PMID- 9326449 TI - Loss of density-dependent growth inhibition and dissociation of alpha-catenin from E-cadherin. AB - Normal human breast epithelial (HBE) cells at early (9th) passage ceased growth and formed a monolayer when they reached confluence. Immunostaining and Western blotting revealed that alpha- and beta-catenins colocalized and coprecipitated with E-cadherin, suggesting a complex formation of E-cadherin with alpha- and beta-catenins in early passage cells. In contrast, HBE cells at late (12-13th) passage did not cease growth after confluence but stratified. The late passage cells exhibited enhanced colony forming ability in soft agar compared with early passage cells, however, they had a definite proliferating lifespan and were primarily diploid. In late passage cells grown as multilayers, alpha-catenin was expressed but did not colocalize or coprecipitate with E-cadherin, suggesting its dissociation from E-cadherin. Coimmunoprecipitation of alpha-actinin with alpha catenin suggested an indirect link between the E-cadherin-beta-catenin complex and alpha-actinin via alpha-catenin in early, but not in late passage cells. Beta Catenin in late passage cells was tyrosine phosphorylated and was not dephosphorylated following the addition of inhibitors of tyrosine kinases. Inhibition of dephosphorylation of beta-catenin in early passage cells by vanadate, an inhibitor of protein tyrosine phosphatases, caused overgrowth of cells beyond the saturation density and loss of alpha-catenin from the E-cadherin beta-catenin complex. The results suggest that E-cadherin requires its association with alpha-actinin-associated alpha-catenin to maintain epithelial monolayers and accomplish the density-dependent inhibition of growth. In addition, association between E-cadherin and alpha-catenin is suggested to be prevented by the presence of tyrosine phosphorylated beta-catenin which associates with E-cadherin. PMID- 9326450 TI - Effect of strain on human keratinocytes in vitro. AB - Tissue expansion, a technique to enlarge the skin surface area with an expandable balloon, has been widely used in reconstructive surgery. Although the effect of tissue expansion on in vivo skin physiology and histology has been well documented, it remains unclear whether keratinocytes or other cell types are responsible for these changes. Therefore, we investigated the in vitro effect of cyclic (10 cycles/min, 150 mmHg) or constant (continuous, 150 mmHg) strain on human keratinocyte phenotype and relevant mechanosignaling pathways. Our results demonstrate that keratinocytes subjected to cyclic strain exhibit a significant (P < 0.05) increase in cell proliferation (49.2+/-15.8%), DNA synthesis (37.7+/ 4.5%), elongation (20.3+/-2.7%), and protein synthesis (17.9+/-6.6% increase) as compared with stationary controls. In contrast, keratinocytes subjected to constant strain were unaffected aside from a modest transitory increase in the proliferative rate. Keratinocytes subjected to cyclic strain aligned perpendicular to the force vector (24.2+/-1.6 degrees) as compared with stationary controls (40.4+/-2.2 degrees; the smaller degree indicates better alignment). We also report strain-induced reduction in the levels of cyclic adenosine mono phosphate (cAMP), protein kinase A (PKA), and prostaglandin E2 (PGE2) as compared with stationary controls (cAMP, 30+/-7.5%; PKA, 45+/-17%; PGE2, 58+/-4.3%; percent decrease vs. that of control). We conclude that direct application of cyclic strain on human keratinocytes modulates cell phenotype and cAMP-mediated signaling pathways in an inverse manner. Moreover, keratinocytes may play an important role in previously observed alterations in skin properties associated with tissue expansion and other strain-induced responses. PMID- 9326451 TI - Rapid elevation of neuronal cytoplasmic calcium by apolipoprotein E peptide. AB - Apolipoprotein E (apoE) and certain peptides derived from it have been shown to exert neurotoxic effects in vitro, and apoE has been linked to the etiology of Alzheimer's disease. The mechanisms underlying these toxic and pathological effects are, however, not known. To approach this question, we have studied the effects of apoE peptides on the cytoplasmic calcium ([Ca2+]i) homeostasis of cultured cortical neurons. A tandem dimer repeat peptide (apoEdp) derived from the receptor binding domain of apoE was found to have a potent effect on elevation of [Ca2+]i calcium. The pathway by which apoEdp exerted this effect was shown to involve both the mobilization of intracellular calcium and the influx of extracellular calcium, although the effect on influx was more pronounced. Calcium mobilization occurs via a G-protein-linked phospholipase C (PLC) pathway, whereas calcium influx appears to involve a novel Co2+-sensitive channel. Both the mobilization and the influx of calcium require the binding of the apoE peptide to a membrane receptor because both pathways are blocked by antibody to low-density lipoprotein receptor-related protein. The data suggest that the neurotoxic effects of apoE may be mediated by a persistent elevation of [Ca2+]i. PMID- 9326452 TI - Interleukin-1 induces major phenotypic changes in human skin microvascular endothelial cells. AB - To determine the role of the pleiotropic cytokine interleukin-1 (IL-1) on the activation of endothelial cells during inflammation and angiogenesis, pure populations of human dermal microvascular endothelial cells (HDMEC) were obtained by immunoaffinity purification using Ulex europaeus agglutinin-1 and platelet endothelial cell adhesion molecule-1 (PECAM-1) antibody. Exposure of HDMEC to IL 1beta induced morphologic and physiologic changes characterized by 1) phenotypic modulation of endothelial cells from epithelioid to spindle-shaped cells accompanied by reorganization of vimentin filaments; 2) gradual decrease to a complete absence of the endothelial cell markers von Willebrand factor (vWf) and PECAM-1; and 3) increased capability to form tubule-like structures when overlaid with collagen gels. The IL-1 effect on cell morphology, growth, and decrease of endothelial cell antigens was potentiated by basic fibroblast growth factor (bFGF). Similar results were observed in mitotically arrested gamma-irradiated cells demonstrating that the spindle-shaped cells observed after IL-1 stimulation were derived from epithelioid endothelial cells and that DNA synthesis was not required to effect these changes. Immunostaining with an antibody specific for human fibroblasts was negative and further confirmed the endothelial cell origin of the spindle-shaped cells. These data demonstrate that IL-1 can induce phenotypic changes in HDMEC from epithelioid to spindle-shaped, mesenchymal-like cells, that these cells are more susceptible to stimulation by bFGF, and that they lose biochemical and functional properties characteristic of epithelioid HDMEC. PMID- 9326453 TI - Dependence of mesenchymal cell responses on duration of exposure to bone morphogenetic protein-2 in vitro. AB - Bone morphogenetic proteins (BMPs) induce osteoblastic responses in cultures of pluripotent mesenchymal cells. The effects of chronic treatment of these cells with BMPs and of withdrawal following exposure, however, have not been fully elucidated. Thus, the aim of this study was to obtain information about the duration of exposure to recombinant human BMP-2 (rhBMP-2) required for expression and retention of osteoblastic characteristics with subsequent formation of a mineralized extracellular matrix in mesenchymal cell cultures. C3H1OT1/2 cells and bone marrow stromal cells were cultured with 1 mug/ml rhBMP-2 for either 0, 7, 14, 21, or 28 days, with the remainder of the 4 week total culture period in the absence of rhBMP-2. Growth and expression of osteoblastic characteristics were examined at the end of each week. C3H1OT1/2 cells responded to increasing duration of exposure to rhBMP-2 with increased cell growth. Additionally, the longer the cells were exposed to rhBMP-2, the more fully they expressed and sustained osteoblastic traits, i.e., they exhibited duration of exposure dependent higher levels of alkaline phosphatase and osteocalcin and larger total amounts of mineral in the matrix. In comparison, exposure of bone marrow stromal cells to rhBMP-2 for at least 14 days restrained cell growth and prevented detachment. With respect to osteoblastic traits, stromal cells exposed to rhBMP-2 also exhibited a dependence on the duration of exposure, however, cultures treated for 14, 21, or 28 days exhibited similar levels of alkaline phosphatase activity and comparable amounts of calcium in the mineralizing matrix. PMID- 9326454 TI - Extracellular matrix heparan sulfate modulates endothelial cell susceptibility to Staphylococcus aureus. AB - The ability of extracellular matrix heparan sulfate to alter the susceptibility of human endothelial cells to S. aureus was investigated. Endothelial cells grown on extracellular matrix synthesized by S. aureus-infected endothelial cells were more susceptible to subsequent staphylococcal infection than endothelial cells grown on the extracellular matrix synthesized by untreated endothelial cells. Endothelial cells were more susceptible to S. aureus infection when 1) grown on heparitinase-treated extracellular matrix that removed heparan sulfate chains, 2) grown on extracellular matrix produced by chlorate-treated endothelial cells that reduced sulfation in the matrix heparan sulfate proteoglycans, 3) grown on heparan sulfate purified from extracellular matrix elaborated by infected endothelial cells, and 4) endothelial cells were chlorate-treated and therefore expressed desulfated cellular heparan sulfate proteoglycans. Extracellular matrix produced by S. aureus-infected endothelial cells contained heparan sulfate proteoglycans with reduced sulfation. The altered extracellular matrix with reduced sulfated heparan sulfate proteoglycans signalled the uninfected endothelial cells to produce under sulfated cellular heparan sulfate proteoglycans that increased S. aureus adherence to the endothelial cells. PMID- 9326455 TI - Interferon-gamma-induced JAK2 and STAT1 signalling in a human salivary gland cell line. AB - We have used a human salivary gland cell line (HSG) as a possible in vitro model to evaluate the effects of IFN-gamma on human salivary gland epithelium (Wu et al., 1994, 1996, 1997). In the present study, we examined the JAK-STAT signal transduction pathway in IFN-gamma-treated HSG cells. We demonstrate that JAK2 and Stat1 are phosphorylated at tyrosine residues in a time- and concentration dependent manner following exposure to IFN-gamma. In addition, we show that activation of this signalling pathway is decreased by the addition of a blocking antibody to the IFN-gamma receptor. The same maneuver is also able to reduce by approximately 50-70% the surface expression of two IFN-gamma-induced immunoregulatory molecules: HLA-DR and ICAM-1. These results demonstrate that the JAK2 and Stat1 signalling pathway is active in salivary-derived epithelial cells and may contribute to their immunopathologic destruction. PMID- 9326456 TI - A piece of my mind. Be careful what you wish for. PMID- 9326457 TI - 'HIV specialists': the time has come. PMID- 9326458 TI - Management of patients with HIV/AIDS. Who should care? PMID- 9326459 TI - Thalidomide back--under strict control. PMID- 9326460 TI - New focus placed on von Willebrand disease. PMID- 9326461 TI - 1997 Albert Lasker and Gairdner Foundation International Medical Research awardees named. PMID- 9326462 TI - From the Centers for Disease Control and Prevention. State-specific birth rates for teenagers--United States, 1990-1996. PMID- 9326463 TI - From the Centers for Disease Control and Prevention. Use of rollover protective structures--Iowa, Kentucky, New York, and Ohio, 1992-1997. PMID- 9326464 TI - From the Centers for Disease Control and Prevention. Update: Staphylococcus aureus with reduced susceptibility to vancomycin--United States, 1997. PMID- 9326465 TI - Contempo 1997: obstetrics and gynecology. PMID- 9326466 TI - Contempo 1997: dermatology. PMID- 9326467 TI - Contempo 1997: dermatology. PMID- 9326468 TI - Contempo 1997: economics. PMID- 9326469 TI - Contempo 1997: economics. PMID- 9326470 TI - Contempo 1997: the face of medicine. PMID- 9326471 TI - Contempo 1997: end-of-life care. PMID- 9326472 TI - Contempo 1997: family medicine. PMID- 9326473 TI - Pediatric window-cord strangulations. PMID- 9326474 TI - Affirmative action and other special consideration admissions at the University of California, Davis, School of Medicine. AB - CONTEXT: The use of race as a criterion for admission to medical schools and other professional schools has become increasingly controversial. This study documents the experience of students at one medical school, admitted through a special admissions process that included race as one consideration. OBJECTIVE: To examine the medical school, postgraduate training, and career experiences of students admitted by a special consideration admission program that included traditional affirmative action admissions. DESIGN: Twenty-year, retrospective, matched-cohort study. SETTING: A public medical school. STUDY POPULATION: All affirmative action and other special consideration admissions between 1968 and 1987 (20 years). MAIN OUTCOME MEASURES: Academic progress, national board examination scores, graduation, residency evaluations, and practice characteristics. RESULTS: During the study period, 20% of students were special consideration admissions (range, 10%-45% per year). Of special consideration admissions, 53.5% were minority students, while 19% of regular admissions were minority students. When only underrepresented minority groups are analyzed, 42.7% of special consideration admissions and 4.0% of regular admissions were minorities. Of special consideration admissions, 94% graduated vs 97% of regular admissions. Regular admission students were more likely to receive honors or an A grade on core basic and clinical science courses. There was no difference in failure rates of core courses. Regular admission students had higher scores on Parts I and II of the National Board of Medical Examiners examination, and special consideration students were more likely to repeat the examination to receive a passing grade. Following graduation, the experience of the special consideration admission students was very similar to that of regular admission students. There was no difference in completion of residency training or evaluation of performance by residency directors. Both populations selected primary care disciplines at the same rate. The practice characteristics of the 2 populations were remarkably similar. CONCLUSIONS: Criteria other than undergraduate grade point average and Medical College Admission Test scores can be used in predicting success in medical school. An admissions process that allows for ethnicity and other special characteristics to be used heavily in admission decisions yields powerful effects on the diversity of the student population and shows no evidence of diluting the quality of the graduates. PMID- 9326475 TI - Patterns of tuberculosis transmission in Central Los Angeles. AB - CONTEXT: Recent studies suggest that many tuberculosis cases in urban areas result from recent transmission. Delineation of the epidemiologic links between patients is important to optimize strategies to reduce tuberculosis transmission. OBJECTIVE: To identify epidemiologic links among recently infected urban patients with tuberculosis. DESIGN: Prospective evaluation of patients with tuberculosis. SETTING: Central Los Angeles, Calif. PATIENTS: A total of 162 patients who had culture-proven tuberculosis. INTERVENTIONS: Patients were prospectively interviewed to identify their contacts and whereabouts. The IS6110-based and pTBN12-based restriction fragment length polymorphism analyses were performed on Mycobacterium tuberculosis isolates. Patients whose isolates had identical or closely related restriction fragment length polymorphism patterns were considered a cluster. Unconditional logistic regression was used to identify independent predictors of clustering. MAIN OUTCOME MEASURES: Relationship of clinical and epidemiologic variables to clustering. RESULTS: A total of 96 (59%) of 162 patients were in 8 clusters. Only 2 of the 96 clustered patients named others in the cluster as contacts. The degree of homelessness was an independent predictor of clustering. Compared with nonclustered patients, patients in 6 clusters were significantly more likely to have spent time at 3 shelters and other locations when at least 1 patient in the cluster was contagious, and these locations were independent predictors of clustering. Among nonhomeless persons, clustered patients were significantly more likely than nonclustered patients to have used daytime services at 3 shelters. CONCLUSIONS: (1) Traditional contact investigation does not reliably identify patients infected with the same M tuberculosis strain, and (2) locations at which the homeless congregate are important sites of tuberculosis transmission for homeless and nonhomeless persons. Measures that reduce tuberculosis transmission should be based on locations rather than on personal contacts. PMID- 9326476 TI - Use of standardized patients to assess between-physician variations in resource utilization. AB - CONTEXT: As medical costs are increasingly being scrutinized, there is heightened interest in defining variations in physician behavior in clinical settings. OBJECTIVE: To evaluate if standardized patient (SP) technology is a reliable and feasible method of studying interphysician variations in test ordering, referral requests, prescribing behavior, and visit costs. DESIGN: The study was conducted with blinded SP visits to family medicine and internal medicine residents, university-affiliated family physicians, and community-based family physicians. Resource utilization and visit costs were assessed using test requisitions, consult requests, and prescriptions that were collected by the SPs. SETTING: Physicians' offices in ambulatory care, hospital-based clinics and in the community. PARTICIPANTS: Four persons (aged 57-77 years) trained to simulate having osteoarthritis of the hip. In one simulation, the patient had gastropathy due to nonsteroidal anti-inflammatory drug use, and in the other, the patient sought therapy for hip discomfort. MAIN OUTCOME MEASURES: Reliability of cost estimates of physician services, tests, consultations, prescriptions, and total visits and test-ordering behavior for nonsteroidal anti-inflammatory gastropathy. RESULTS: Overall, 112 (63%) of the physicians who were sent invitations to the study agreed to participate. Of 312 total SP visits conducted over a 1-year period, unblinding due to SP detection occurred on 36 occasions (11.5%). Reliable cost estimates of physician services, tests, and consultations, and moderately reliable estimates of total visit costs, were obtained with 4 visits per practicing physician and with 2 visits per resident. There were extreme variations in total visit costs generated by the study physicians. A small number of physicians had a major impact on this variability. CONCLUSION: Standardized patient technology provides a reliable, feasible method to assess variations in resource utilization between physicians. PMID- 9326477 TI - Endothelial activation and development of coronary artery disease in transplanted human hearts. AB - CONTEXT: The development of coronary artery disease in heart transplants is often associated with graft failure. Early detection of allografts prone to develop this disease is essential to institute new therapeutic approaches that could prolong allograft function. OBJECTIVE: To determine if early activation of arterial/arteriolar endothelium predicts the development of coronary artery disease, graft failure, or both in transplanted human hearts. DESIGN: Prospective cohort study. SETTING: Heart Transplant Center. PARTICIPANTS: A total of 121 consecutive adult cardiac allograft recipients who received transplants between 1988 and 1995 and were followed up through 1996. MAIN OUTCOME MEASURES: Development of coronary artery disease and graft failure. METHODS: Immunocytochemistry was performed on serial endomyocardial biopsy specimens to evaluate endothelial activation markers (intercellular adhesion molecule-1 and histocompatibility antigen HLA-DR) in arteries and arterioles. The presence and progression of coronary artery disease was evaluated by annual coronary angiograms with side-by-side comparisons. RESULTS: None of the 121 donor hearts showed arterial/arteriolar endothelial activation before transplantation. Arterial/arteriolar endothelial activation was present in 78 and absent in 43 of 121 allografts during the first 3 months after transplantation. The time of appearance and the proportion of biopsy specimens showing endothelial activation during these first 3 months were significantly associated with the risk of developing coronary artery disease, the progression of the disease, and the time required to develop the disease (P<.001). Significantly more patients with arterial/arteriolar endothelial activation died or received a second transplant (P<.001). CONCLUSIONS: Activation of arterial/arteriolar endothelium in transplanted human hearts predicts development of coronary artery disease and increased risk of graft failure. PMID- 9326478 TI - Hair loss after routine immunizations. AB - CONTEXT: Alopecia is a recognized adverse effect of numerous medications, but vaccines are not normally considered a cause for unexpected loss of hair. OBJECTIVE: To describe case reports of hair loss after routine vaccines and to assess the hypothesis that vaccinations might induce hair loss. DESIGN: Case series with telephone follow-up. METHODS: Review of spontaneous reports to the Food and Drug Administration, the Centers for Disease Control and Prevention, and the Vaccine Adverse Event Reporting System. MAIN OUTCOME MEASURE: Loss of hair following immunization. RESULTS: A total of 60 evaluable reports submitted since 1984 and coded for "alopecia" after immunizations included 16 with positive rechallenge (hair loss after vaccination on more than 1 occasion), 4 of which were definite and 12 possible or probable. Of the 60 cases, 46 had received hepatitis B vaccines. Both of the currently available recombinant products, as well as the former plasma-derived product, were represented. Females predominated in all age groups. The majority of patients recovered, but clinical features, such as intervals from vaccination until onset and the extent and reversibility of hair loss, varied widely. Nine patients reported previous medication allergy. CONCLUSION: There may be an association, probably very rare, between vaccinations and hair loss. More than 1 pathophysiologic mechanism may be responsible. Since apparently nonrandom distributions by vaccine, age, and sex could reflect biased case ascertainment, further research will be needed in defined populations with consistent case detection. PMID- 9326479 TI - A 56-year-old woman with chronic fatigue syndrome. PMID- 9326480 TI - A 96-year-old woman with insomnia, 1 year later. PMID- 9326481 TI - Diagnosis and treatment of depression in late life. Consensus statement update. AB - OBJECTIVE: To reexamine the conclusions of the 1991 National Institutes of Health Consensus Panel on Diagnosis and Treatment of Depression in Late Life in light of current scientific evidence. PARTICIPANTS: Participants included National Institutes of Health staff and experts drawn from the Planning Committee and presenters of the 1991 Consensus Development Conference. EVIDENCE: Participants summarized relevant data from the world scientific literature on the original questions posed for the conference. PROCESS: Participants reviewed the original consensus statement and identified areas for update. The list of issues was circulated to all participants and amended to reflect group agreement. Selected participants prepared first drafts of the consensus update for each issue. All drafts were read by all participants and were amended and edited to reflect group consensus. CONCLUSIONS: The review concluded that, although the initial consensus statement still holds, there is important new information in a number of areas. These areas include the onset and course of late-life depression; comorbidity and disability; sex and hormonal issues; newer medications, psychotherapies, and approaches to long-term treatment; impact of depression on health services and health care resource use; late-life depression as a risk factor for suicide; and the importance of the heterogeneous forms of depression. Depression in older people remains a significant public health problem. The burden of unrecognized or inadequately treated depression is substantial. Efficacious treatments are available. Aggressive approaches to recognition, diagnosis, and treatment are warranted to minimize suffering, improve overall functioning and quality of life, and limit inappropriate use of health care resources. PMID- 9326482 TI - Who is still uninsured in Minnesota? Lessons from state reform efforts. AB - OBJECTIVE: To describe Minnesota's health care system reform efforts and their implications for other state and national reform initiatives, document the rate of uninsurance in 1990 and 1995 with special attention to childrens' access to health insurance, and examine the effectiveness of MinnesotaCare, a voluntary state-subsidized health care plan, in serving its target population. DESIGN: Three cross-sectional telephone surveys: 2-stage random samples of Minnesotans of all ages in 1990 and 1995 and a stratified random sample of MinnesotaCare enrollees in 1994. PARTICIPANTS: For the 2 statewide surveys, 10310 respondents participated in 1990 and 11519 in 1995; more detailed information was collected on approximately 1600 respondents in each survey. Eight hundred MinnesotaCare enrollees participated in the third survey conducted in 1994. MAIN OUTCOME MEASURE: Changes in rates of uninsurance. RESULTS: While the rate of uninsurance increased at the national level, the point-in-time Minnesota rate remained stable and low at 6% between 1990 and 1995. The proportion of children uninsured for 12 months or more decreased from 5.2% in 1990 to 3.1% in 1995, while the proportion of uninsured single adults remained stable at approximately 11%. There was no evidence that MinnesotaCare enrollees are gaming the program, or that the program has resulted in significant erosion from the private market. CONCLUSIONS: MinnesotaCare has enabled the state to maintain a low rate of uninsurance and has reduced this rate among its primary target: children. The program has been less effective in enrolling single adults, although it may be too early to witness the effects of recent expansions targeting this group. Minnesota's experience suggests that other state and national reform efforts aimed at reducing uninsurance, particularly among children, are likely to be successful. PMID- 9326483 TI - Medical school admission criteria. The needs of patients matter. PMID- 9326484 TI - Endomyocardial biopsies. An early warning system for chronic transplant arteriopathy. PMID- 9326485 TI - eIF4G: translation's mystery factor begins to yield its secrets. PMID- 9326486 TI - A guide to RNA editing. PMID- 9326487 TI - Poly (rC) binding protein 2 forms a ternary complex with the 5'-terminal sequences of poliovirus RNA and the viral 3CD proteinase. AB - Poly(rC) binding protein 2 (PCBP2) forms a specific ribonucleoprotein (RNP) complex with the 5'-terminal sequences of poliovirus genomic RNA, as determined by electrophoretic mobility shift assay. Mutational analysis showed that binding requires the wild-type nucleotide sequence at positions 20-25. This sequence is predicted to localize to a specific stem-loop within a cloverleaf-like secondary structure element at the 5'-terminus of the viral RNA. Addition of purified poliovirus 3CD to the PCBP2/RNA binding reaction results in the formation of a ternary complex, whose electrophoretic mobility is further retarded. These properties are consistent with those described for the unidentified cellular protein in the RNP complex described by Andino et al. (Andino R, Rieckhof GE, Achacoso PL, Baltimore D, 1993, EMBO J 12:3587-3598). Dicistronic RNAs containing mutations in the 5' cloverleaf-like structure of poliovirus that abate PCBP2 binding show a decrease in RNA replication and translation of gene products directed by the poliovirus 5' noncoding region in vitro, suggesting that the interaction of PCBP2 with these sequences performs a dual role in the virus life cycle by facilitating both viral protein synthesis and initiation of viral RNA synthesis. PMID- 9326488 TI - Functional groups on the cleavage site pyrimidine nucleotide are required for stabilization of the hammerhead transition state. AB - The role of individual functional groups on cytidine 17 in the hammerhead ribozyme was assessed by introducing modified pyrimidines into two kinetically well-characterized hammerheads. As long as the pyrimide ring size was maintained, the modifications had no effect on substrate binding, suggesting that the C17-C3 hydrogen bond observed in the X-ray structure is energetically neutral. However, modification of the exocyclic amino group and the carbonyl of C17 reduced the cleavage rate significantly, indicating that these groups are important in stabilizing the transition-state structure. C17 modifications did not affect the ratio of the forward and reverse reaction rates. Thus, unlike that believed previously, C17 is another one of many hammerhead residues critical in maintaining its active structure. PMID- 9326489 TI - The yeast Prp3 protein is a U4/U6 snRNP protein necessary for integrity of the U4/U6 snRNP and the U4/U6.U5 tri-snRNP. AB - Previously, yeast prp3 mutants were found to be blocked prior to the first catalytic step of pre-mRNA splicing. No splicing intermediates or products are formed from pre-mRNA in heat-inactivated prp3 mutants or prp3 mutant extracts. Here we show that Prp3p is a component of the U4/U6 snRNP and is also present in the U4/U6.U5 tri-snRNP. Heat inactivation of prp3 extracts results in depletion of free U6 snRNPs and U4/U6.U5 tri-snRNPs, but not U4/U6 snRNPs or U5 snRNPs. Free U4 snRNP, normally not present in wild-type extracts, accumulates under these conditions. Assays of in vivo levels of snRNAs in a prp3 mutant revealed that amounts of free U6 snRNA decreased, free U4 snRNA increased, and U4/U6 hybrids decreased slightly. These results suggest that Prp3p is required for formation of stable U4/U6 snRNPs and for assembly of the U4/U6.U5 tri-snRNP from its component snRNPs. Upon inactivation of Prp3p, spliceosomes cannot assemble from prespliceosomes due to the absence of intact U4/U6.U5 tri-snRNPs. Prp3p is homologous to a human protein that is a component of U4/U6 snRNPs, exemplifying the conservation of splicing factors between yeast and metazoans. PMID- 9326490 TI - In vivo HIV-1 frameshifting efficiency is directly related to the stability of the stem-loop stimulatory signal. AB - In many retroviruses, the expression of reverse transcriptase, protease, and integrase is dependent upon a -1 frameshift event. The frameshift signal is composed of a slippery sequence where the ribosome shifts, and a downstream stimulatory sequence. In most cases, the stimulatory sequence is a pseudoknot, but in some viruses, such as human immunodeficiency virus type 1 (HIV-1), a single stem-loop is involved. Here, we analyzed the precise role of the stem-loop thermodynamic stability. We tested the frameshifting stimulatory activity of a series of HIV-1-derived sequences showing a stepwise increment of the estimated deltaG degrees. These sequences were introduced at the junction of a lacZ-luc fusion gene cloned on a versatile expression vector, and the different constructs were tested in Saccharomyces cerevisiae and in mouse NIH3T3 cells. The results showed that the frameshifting efficiency was correlated directly to the stem stability between deltaG degrees = -2.5 kcal mol(-1) and deltaG degrees = -19.4 kcal mol(-1). This demonstrates the essential role of the stability of the stem loop and does not support the involvement of a specific RNA-binding protein target sequence. However, increasing further the stem stability led to a diminution of frameshifting efficiency, suggesting that the stem-loop acts through a precise kinetic of pausing. Because the same pattern was observed in both yeast and mouse cells, it is likely that the stimulatory mechanism is conserved through evolution. PMID- 9326491 TI - Probing the structure of the regulatory region of human transferrin receptor messenger RNA and its interaction with iron regulatory protein-1. AB - A portion of the 3'UTR of the human transferrin receptor mRNA mediates iron dependent regulation of mRNA stability. The minimal RNA regulatory region contains three conserved hairpins, so-called iron responsive elements (IREs), that are recognized specifically by iron regulatory proteins (IRPs). The structure of this regulatory region and its complex with IRP-1 was probed using a combination of enzymes and chemicals. The data support the existence of an intrinsic IRE loop structure that is constrained by an internal C-G base pair. This particular structure is one of the determinants required for optimal IRP binding. IRP-1 covers one helical turn of the IRE and protects conserved residues in each of the three IREs: the bulged cytosine and nucleotides in the hairpin loops. Two essential IRP-phosphate contacts were identified by ethylation interference. Three-dimensional modeling of one IRE reveals that IRP-1 contacts several bases and the ribose-phosphate backbone located on one face in the deep groove, but contacts also exist with the shallow groove. A conformational change of the IRE loop mediated by IRP-1 binding was visualized by Pb2+-catalyzed hydrolysis. This effect is dependent on the loop structure and on the nature of the closing base pair. Within the regulatory region of transferrin receptor mRNA, IRP-1 induces reactivity changes in a U-rich hairpin loop that requires the presence of the stem-loop structure located just downstream the endonucleolytic cleavage site identified by Binder et al. (Binder R et al. 1994, EMBO J 13:1969 1980). These results provide indications of the mechanism by which IRP-1 stabilizes the transferrin receptor mRNA under iron depletion conditions. PMID- 9326492 TI - Mammalian prothymosin alpha links to tRNA in Escherichia coli cells. AB - Prothymosin alpha, a small and highly acidic nuclear protein related to cell proliferation, is known to be covalently attached to a small unidentified cytoplasmic RNA in mammalian cells. Here we demonstrate that recombinant rat prothymosin a links covalently to an RNA when overproduced in Escherichia coli cells. The RNA species of this complex is represented by a wide range of bacterial tRNAs. tRNA(Lys), tRNA(3Ser), tRNA(2Ile), and tRNA(mMet) were identified by sequencing. Prothymosin alpha appears to be linked to the 5' terminus of tRNA. tRNA attachment site lies close to the carboxy-terminus of prothymosin alpha. Furthermore, the carboxy-terminal peptide of prothymosin alpha is also competent for tRNA binding. The site of tRNA attachment coincides with the nuclear localization signal of prothymosin alpha, and tRNA binding might be expected to affect subcellular localization of this protein. PMID- 9326493 TI - Substrate structure requirements of the Pac1 ribonuclease from Schizosaccharmyces pombe. AB - The Pac1 ribonuclease of Schizosaccharomyces pombe is a member of the RNase III family of double-strand-specific ribonucleases. To examine RNA structural features required for efficient cleavage by the Pac1 RNase, we tested a variety of double-stranded and hairpin RNAs as substrates for the enzyme. The Pac1 RNase required substrates that have a minimal helix length of about 20 base pairs. The enzyme cut both strands of the helix at sites separated by two base pairs. However, Pac1 was also able to make a single-stranded cleavage within an internal bulge of an authentic Escherichia coli substrate at the same site chosen by RNase III. Pac1 efficiently degraded the structurally complex adenovirus VA RNA(I), but was inactive against the short HIV-1 TAR RNA hairpin. These results indicate that the Pac1 RNase prefers straight, perfect helices, but it can tolerate internal bulges that do not distort the helix severely. Like its homologue from Saccharomyces cerevisiae, the Pac1 RNase cleaved at two in vivo RNA processing sites in a hairpin structure in the 3' external transcribed spacer of the S. pombe pre-rRNA, suggesting a role for the enzyme in rRNA maturation. PMID- 9326494 TI - Identification of critical nucleotide positions for plastid RNA editing site recognition. AB - Transcripts in higher plant cell organelles undergo RNA editing by C-to-U conversion. Both the mechanistic steps and the factors involved in this process are largely unknown. To gain a better understanding of the molecular interactions involved in organellar RNA editing, we have begun to identify critical nucleotide positions for plastid RNA editing-site recognition. We performed a scanning point mutagenesis on a sequence motif separating editing sites IV and V in the tobacco ndhB transcript. The constructs were integrated into the chloroplast genome by the biolistic process and the effect of each point mutation on editing of both the upstream and the downstream site was measured. In addition to a previously identified sequence element located upstream of both sites, only few nucleotide positions 5' and 3' of an editing site turned out to be of critical importance. Unexpectedly, our study revealed that mutation of the upstream site leads to loss of editing at the downstream site. However, our results also indicate that, even though closely adjacent editing sites can share common recognition elements in cis, they are edited independently and not in a polar fashion. PMID- 9326495 TI - Possible involvement of somatolactin in the regulation of plasma bicarbonate for the compensation of acidosis in rainbow trout AB - Somatolactin is a putative pituitary hormone of the growth hormone/prolactin family in fish. Its function is still unknown. The effects of environmental hypercapnia and hypoxia, acid (HCl) infusion and exhaustive exercise on plasma somatolactin levels were examined in the chronically cannulated rainbow trout to study the possible physiological roles of somatolactin. Respiratory acidosis induced by hypercapnia (2% CO2) did not affect plasma somatolactin level. In contrast, metabolic acidosis induced by acid infusion and exercise increased plasma somatolactin level. Blood pH was depressed to a similar extent by both types of acidosis, whereas plasma [HCO3-] was elevated by respiratory acidosis but reduced by metabolic acidosis. A moderate hypoxia (water PO2 9.3kPa) affected neither acid­base status nor plasma somatolactin level. A more severe hypoxia (water PO2 6.1kPa) resulted in metabolic acidosis accompanied by an apparent rise in plasma somatolactin level, although the difference in somatolactin level from the control value was not statistically significant. Somatolactin immunoneutralization retarded recovery of plasma [HCO3-] following acid infusion. These results indicate that somatolactin is involved in the retention of HCO3- during metabolic acidosis but not in the active accumulation of HCO3- for acid­base compensation of respiratory acidosis in rainbow trout Oncorhynchus mykiss. PMID- 9326496 TI - Imidacloprid actions on insect neuronal acetylcholine receptors AB - The neonicotinoid insecticide Imidacloprid acts at three pharmacologically distinct acetylcholine receptor (AChR) subtypes in the cockroach (Periplaneta americana) nervous system, but is ineffective on muscarinic receptors. Imidacloprid (3-100µmoll-1) induced dose-dependent depolarizations at cockroach cercal afferent/giant interneurone synapses. These responses were insensitive to 20µmoll-1 atropine but were completely blocked by the nicotinic antagonist mecamylamine (50µmoll-1). Similarly, Imidacloprid induced depolarizations of cultured cockroach dorsal unpaired median (DUM) neurones dissociated from the same (terminal abdominal) ganglion were also completely blocked by 100µmoll-1 mecamylamine. However, two components of the response could be distinguished on the basis of their differential sensitivities to 0.1µmoll-1-bungarotoxin (-BTX), which selectively blocks AChRs with 'mixed' nicotinic/muscarinic pharmacology in this preparation. This indicates that Imidacloprid affects both AChRs sensitive to -BTX and -BTX insensitive nicotinic acetylcholine receptors (nAChRs). Thus, in the cockroach, Imidacloprid activates -BTX-sensitive synaptic nAChRs in giant interneurones, BTX-insensitive extrasynaptic nAChRs in DUM neurones, and a recently characterized DUM neurone 'mixed' AChR that is sensitive to both nicotinic and muscarinic ligands. Imidacloprid does not act on muscarinic acetylcholine receptors (mAChRs) present on DUM neurone cell bodies and at the cercal afferent/giant interneurone synapses. This study shows that Imidacloprid can act on pharmacologically diverse nAChR subtypes. PMID- 9326497 TI - Measuring the angle of attack of beating insect wings: robust three-dimensional reconstruction from two-dimensional images AB - A robust technique for determining the angle of attack of insect wings from film of free flight has to date proved elusive. This report describes the development of two new methods ­ the Strips and Planes techniques ­ which were designed to overcome some of the limitations experienced in previous studies. The accuracy and robustness of these novel methods were tested extensively using simulated hawkmoth wing outlines generated for a realistic range of wing positions and torsion. The results were compared with those from two existing methods ­ the Symmetry and Landmarks procedures. The performance of the latter technique will be strongly species-dependent; it could not be successfully applied to hawkmoth flight because of practical difficulties in obtaining suitable landmarks. The Planes method was the least successful of the remaining techniques, especially during those phases of the wingbeat when the orientations of the two wings relative to the camera viewpoint were similar. The Symmetry and Strips methods were tested further to investigate the effects on their performance of introducing additional camber or wing motion asymmetry. The results showed clearly that the Strips method should be the technique of choice wherever the axis of wing torsion is close to the longitudinal axis of the wing. The procedure involves the experimenter matching a model wing divided into chordwise strips to the true wing outline digitized from high-speed film. The use of strips rather than the points digitized in previous studies meant that the analysis required only one wing outline to be digitized. Symmetry of motion between the left and right wings is not assumed. The implications of requiring human input to the Strips method, as opposed to the strictly numerical algorithms of the alternative techniques, are discussed. It is argued that the added flexibility that this provides in dealing with images which have typically been recorded in suboptimal conditions outweighs the introduction of an element of subjectivity. Additional observations arising from the use of the Strips analysis with high-speed video sequences of hawkmoth flight are given. PMID- 9326502 TI - Feeding mechanism and functional morphology of the jaws of the lemon shark Negaprion brevirostris (Chondrichthyes, Carcharhinidae) AB - This study tests the hypothesis that preparatory, expansive, compressive and recovery phases of biting behavior known for aquatically feeding anamniotes are conserved among extant elasmobranch fishes. The feeding mechanism of the lemon shark Negaprionbrevirostris is examined by anatomical dissection, electromyography and high-speed video analysis. Three types of feeding events are differentiated during feeding: (1) food ingestion primarily by ram feeding; (2) food manipulation; and (3) hydraulic transport of the food by suction. All feeding events are composed of the expansive, compressive and recovery phases common to aquatically feeding teleost fishes, salamanders and turtles. A preparatory phase is occasionally observed during ingestion bites, and there is no fast opening phase characteristic of some aquatically feeding vertebrates. During the compressive phase, palatoquadrate protrusion accounts for 26% of the gape distance during jaw closure and is concurrent with muscle activity in the dorsal and ventral preorbitalis and the levator palatoquadrati. Hydraulic transport events are shorter in duration than ram ingestion bites. Prey ingestion, manipulation and hydraulic transport events are all found to have a common series of kinematic and motor components. Individual sharks are capable of varying the duration and to a lesser extent the onset of muscle activity and, consequently, can vary their biting behavior. We propose a model for the feeding mechanism in carcharhinid sharks, including upper jaw protrusion. This study represents the first electromyographic and kinematic analysis of the feeding mechanism and behavior of an elasmobranch. PMID- 9326504 TI - Mitochondrial arginine kinase in the midgut of the tobacco hornworm (Manduca sexta) AB - Mitochondria isolated from the posterior midgut of the tobacco hornworm (Manduca sexta) contain arginine kinase. The distribution of mitochondrial and cytoplasmic marker enzymes indicates that the presence of mitochondrial arginine kinase is not due to cytoplasmic contamination. Arginine is not oxidized by the midgut mitochondria but, when metabolic substrates and ATP are present, respiration can be initiated by the addition of arginine. Under these conditions, there is no return to State 4 respiration, indicating regeneration of ADP by the arginine kinase reaction. Respiration can be blocked, however, by atractyloside, an inhibitor of the adenine nucleotide translocator. These results indicate that arginine kinase resides outside the matrix. Mitochondrial arginine kinase is specific to l-arginine since analogs of l-arginine are ineffective in stimulating respiration in the presence of ATP. Coupling between the adenine nucleotide translocator and arginine kinase was investigated using kinetic and thermodynamic approaches. There were no differences in the activities of arginine kinase in respiring and non-respiring mitochondria when they were measured at different ATP or arginine concentrations. This result indicates that arginine kinase does not have preferential access to the ATP exported out of the matix. A comparison of the apparent equilibrium constant and the mass action ratio of the arginine kinase reaction also confirms that there is no microcompartmentation of the reaction. PMID- 9326506 TI - The Terminology of "Carcinoma Cell Leukemia" - Is an Alternative Needed? PMID- 9326505 TI - The effects of exposure to ammonia on ammonia and taurine pools of the symbiotic clam AB - The nutrition of the gutless clam Solemyareidi is supported by the activity of intracellular chemoautotrophic bacteria housed in its gill filaments. Ammonia (the sum of NH3 and NH4+) is utilized as a nitrogen source by the association and is abundant in the clam's environment. In the present study, clams were exposed to 0.01­1.3mmoll-1 ammonia for 22­23h in the presence of thiosulfate as a sulfur substrate. Ammonia exposure increased the ammonia concentration in the tissue pools of the gill, foot and visceral mass from 0.5 to 2µmolg 1wetmass, without added ammonia, to as much as 12µmolg-1wetmass in the presence of 0.7 and 1.3mmoll-1 external ammonia. Gill tissue ammonia concentrations were consistently higher than those in the foot and visceral mass. The elevation of tissue ammonia concentration compared with the medium may be due in part to an ammonia trapping mechanism resulting from a lower intracellular pH compared with sea water and greater permeability to NH3 compared with NH4+. Rates of ammonia incorporation into organic matter (assimilation) were determined using 15N as a tracer. 15N-labeled ammonia assimilation was higher in gill than in foot and increased as a function of 15N-labeled ammonia concentration in the medium. The size of the free amino acid (FAA) pool in the gill also increased as a function of ammonia concentration in the medium. This entire increase was accounted for by a single amino acid, taurine, which was the predominant FAA in both gill and foot tissue. Aspartate, glutamate, arginine and alanine were also abundant but their levels were not influenced by external ammonia concentration. Ammonia assimilation appeared to occur at rates sufficient to account for the observed increase in taurine level. These findings suggest that taurine is a major product of ammonia assimilation. PMID- 9326507 TI - Cutaneous T-Cell Lymphomas and Bacterial Superantigens PMID- 9326508 TI - The Effectiveness of Deferiprone in Thalassemia PMID- 9326509 TI - Serum Ig Abnormalities in Mantle Cell Lymphoma PMID- 9326510 TI - Does the Adhesion Molecule CD31 Act as a Minor Histocompatibility Antigen? PMID- 9326511 TI - Serum Transferrin Receptor Levels in Different Stages of Iron Deficiency PMID- 9326512 TI - Congenital Haptoglobin Deficiency PMID- 9326513 TI - Evidence That the RHDVI Deletion Genotype Does Not Exist PMID- 9326514 TI - Hemoglobin E and Pyrimidine 5' Nucleotidase Deficiency PMID- 9326515 TI - Responses of Granulocyte Colony-Stimulating Factor-Mobilized Peripheral Blood Mononuclear Cells to Alloantigen Stimulation PMID- 9326516 TI - Should Patients With Atherosclerosis or Peripheral Vascular Disease Be Stratified for Factor V Leiden? PMID- 9326517 TI - Epstein-Barr Virus Lytic Infection in Lymphocytes and the Persistence of the Virus PMID- 9326574 TI - Broccoli sprouts as inducers of carcinogen-detoxifying enzyme systems: clinical, dietary, and policy implications. PMID- 9326575 TI - Immune responsiveness in vector insects. PMID- 9326576 TI - Wolbachia run amok. PMID- 9326577 TI - An artificial cytochrome P450 that hydroxylates unactivated carbons with regio- and stereoselectivity and useful catalytic turnovers. AB - A catalyst has been synthesized comprising a manganese porphyrin carrying four beta-cyclodextrin groups. It catalyzes the hydroxylation of substrates of appropriate size carrying tert-butylphenyl groups that can hydrophobically bind into the cyclodextrin cavities. In one example as many as 650 catalytic turnovers are seen before the catalyst is oxidatively destroyed, and with a rate comparable to that of typical cytochrome P450 enzymes. In another example, a steroid derivative is regio- and stereoselectively hydroxylated at a single unactivated carbon atom, but more slowly and with fewer turnovers. The carbon attacked is not the most chemically reactive, and the selectivity is determined by the geometry of the catalyst-substrate complex. Nonbinding substrates are not reactive under the conditions used, and substrates with more flexible binding geometries give more than a single product. PMID- 9326579 TI - High-precision 40Ar/39Ar geochronology and the advent of North America's Late Cretaceous terrestrial fauna. AB - A densely sampled, diverse new fauna from the uppermost Cedar Mountain Formation, Utah, indicates that the basic pattern of faunal composition for the Late Cretaceous of North America was already established by the Albian-Cenomanian boundary. Multiple, concordant 40Ar/39Ar determinations from a volcanic ash associated with the fauna have an average age of 98.39 +/- 0.07 million years. The fauna of the Cedar Mountain Formation records the first global appearance of hadrosaurid dinosaurs, advanced lizard (e.g., Helodermatidae), and mammal (e.g., Marsupialia) groups, and the first North American appearance of other taxa such as tyrannosaurids, pachycephalosaurs, and snakes. Although the origin of many groups is unclear, combined biostratigraphic and phylogenetic evidence suggests an Old World, specifically Asian, origin for some of the taxa, an hypothesis that is consistent with existing evidence from tectonics and marine invertebrates. Large-bodied herbivores are mainly represented by low-level browsers, ornithopod dinosaurs, whose radiations have been hypothesized to be related to the initial diversification of angiosperm plants. Diversity at the largest body sizes (>10(6) g) is low, in contrast to both preceding and succeeding faunas; sauropods, which underwent demise in the Northern hemisphere coincident with the radiation of angiosperms, apparently went temporarily unreplaced by other megaherbivores. Morphologic and taxonomic diversity among small, omnivorous mammals, multituberculates, is also low. A later apparent increase in diversity occurred during the Campanian, coincident with the appearance of major fruit types among angiosperms, suggesting the possibility of adaptive response to new resources. PMID- 9326580 TI - Oxidative damage during aging targets mitochondrial aconitase. AB - The mechanisms that cause aging are not well understood. The oxidative stress hypothesis proposes that the changes associated with aging are a consequence of random oxidative damage to biomolecules. We hypothesized that oxidation of specific proteins is critical in controlling the rate of the aging process. Utilizing an immunochemical probe for oxidatively modified proteins, we show that mitochondrial aconitase, an enzyme in the citric acid cycle, is a specific target during aging of the housefly. The oxidative damage detected immunochemically was paralleled by a loss of catalytic activity of aconitase, an enzyme activity that is critical in energy metabolism. Experimental manipulations which decrease aconitase activity should therefore cause a decrease in life-span. This expected decrease was observed when flies were exposed to hyperoxia, which oxidizes aconitase, and when they were given fluoroacetate, an inhibitor of aconitase. The identification of a specific target of oxidative damage during aging allows for the assessment of the physiological age of a specific individual and provides a method for the evaluation of treatments designed to affect the aging process. PMID- 9326581 TI - A yeast sterol auxotroph (erg25) is rescued by addition of azole antifungals and reduced levels of heme. AB - Genetic disruption of the Saccharomyces cerevisiae C-4 sterol methyl oxidase ERG25 gene leads to sterol auxotrophy. We have characterized a suppression system that requires two mutations to restore viability to this disrupted strain. One suppressor mutation is erg11, which is blocked in 14alpha-demethylation of lanosterol and is itself an auxotroph. The second suppressor mutation required is either slu1 or slu2 (suppressor of lanosterol utilization). These mutations are leaky versions of HEM2 and HEM4, respectively; addition of exogenous hemin reverses the suppressing effects of slu1 and slu2. Suppression of erg25 by erg11 slu1 (or erg11 slu2) results in a slow-growing strain in which lanosterol, the first sterol in the pathway, accumulates. This result indicates that endogenously synthesized lanosterol can substitute for ergosterol and support growth. In the triple mutants, all but 1 (ERG6) of the 13 subsequent reactions of the ergosterol pathway are inactive. Azole antibiotics (clotrimazole, ketoconazole, and itraconazole) widely used to combat fungal infections are known to do so by inhibiting the ERG11 gene product, the 14alpha-demethylase. In this investigation, we demonstrate that treatment of the sterol auxotrophs erg25 slu1 or erg25 slu2 with azole antibiotics paradoxically restores viability to these strains in the absence of sterol supplementation via the suppression system we have described. PMID- 9326582 TI - Sphingomyelin depletion in cultured cells blocks proteolysis of sterol regulatory element binding proteins at site 1. AB - The current studies explore the mechanism by which the sphingomyelin content of mammalian cells regulates transcription of genes encoding enzymes of cholesterol synthesis. Previous studies by others have shown that depletion of sphingomyelin by treatment with neutral sphingomyelinase causes a fraction of cellular cholesterol to translocate from the plasma membrane to the endoplasmic reticulum where it expands a regulatory pool that leads to down-regulation of cholesterol synthesis and up-regulation of cholesterol esterification. Here we show that sphingomyelinase treatment of cultured Chinese hamster ovary cells prevents the nuclear entry of sterol regulatory element binding protein-2 (SREBP-2), a membrane-bound transcription factor required for transcription of several genes involved in the biosynthesis and uptake of cholesterol. Nuclear entry is blocked because sphingomyelinase treatment inhibits the proteolytic cleavage of SREBP-2 at site 1, thereby preventing release of the active NH2-terminal fragments from cell membranes. Sphingomyelinase treatment thus mimics the inhibitory effect on SREBP processing that occurs when exogenous sterols are added to cells. Sphingomyelinase treatment did not block site 1 proteolysis of SREBP-2 in 25-RA cells, a line of Chinese hamster ovary cells that is resistant to the suppressive effects of sterols, owing to an activating point mutation in the gene encoding SREBP cleavage-activating protein. In 25-RA cells, sphingomyelinase treatment also failed to down-regulate the mRNA for 3-hydroxy-3-methylglutaryl CoA synthase, a cholesterol biosynthetic enzyme whose transcription depends on the cleavage of SREBPs. Considered together with previous data, the current results indicate that cells regulate the balance between cholesterol and sphingomyelin content by regulating the proteolytic cleavage of SREBPs. PMID- 9326583 TI - D-AKAP2, a novel protein kinase A anchoring protein with a putative RGS domain. AB - Subcellular localization directed by specific A kinase anchoring proteins (AKAPs) is a mechanism for compartmentalization of cAMP-dependent protein kinase (PKA). Using a two-hybrid screen, a novel AKAP was isolated. Because it interacts with both the type I and type II regulatory subunits, it was defined as a dual specific AKAP or D-AKAP1. Here we report the cloning and characterization of another novel cDNA isolated from that screen. This new member of the D-AKAP family, D-AKAP2, also binds both types of regulatory subunits. A message of 5 kb pairs was detected for D-AKAP2 in all embryonic stages and in all adult tissues tested. In brain, skeletal muscle, kidney, and testis, a 10-kb mRNA was identified. In testis, several small mRNAs were observed. Therefore, D-AKAP2 represents a novel family of proteins. cDNA cloning from a mouse testis library identified the full length D-AKAP2. It is composed of 372 amino acids which includes the R binding fragment, residues 333-372, at its C-terminus. Based on coprecipitation assays, the R binding domain interacts with the N-terminal dimerization domain of RIalpha and RIIalpha. A putative RGS domain was identified near the N-terminal region of D-AKAP2. The presence of this domain raises the intriguing possibility that D-AKAP2 may interact with a Galpha protein thus providing a link between the signaling machinery at the plasma membrane and the downstream kinase. PMID- 9326584 TI - Three Ever Shorter Telomere (EST) genes are dispensable for in vitro yeast telomerase activity. AB - Telomerase is a specialized reverse transcriptase consisting of both RNA and protein components. Previous characterization of yeast telomerase function in vivo identified four EST (for ever shorter telomeres) genes that, when mutated, result in the phenotypes expected for a defect in telomerase. Consistent with this genetic prediction, the EST2 gene has recently been shown to encode the catalytic component of telomerase. Using an in vitro assay, we show here that telomerase activity is present in extracts prepared from yeast strains carrying est1-Delta, est3-Delta, and cdc13-2(est) mutations. Therefore, while these three genes are necessary for telomerase function in vivo, they do not encode components essential for core catalytic activity. When Est2p, the one EST gene product found to be essential for catalytic activity, was immunoprecipitated from extracts, the telomerase RNA subunit was also specifically precipitated, supporting the conclusion that these two components are in a stable complex. PMID- 9326585 TI - The cysteine-rich frizzled domain of Frzb-1 is required and sufficient for modulation of Wnt signaling. AB - Convincing evidence has accumulated to identify the Frizzled proteins as receptors for the Wnt growth factors. In parallel, a number of secreted frizzled like proteins with a conserved N-terminal frizzled motif have been identified. One of these proteins, Frzb-1, binds Wnt-1 and Xwnt-8 proteins and antagonizes Xwnt-8 signaling in Xenopus embryos. Here we report that Frzb-1 blocks Wnt-1 induced cytosolic accumulation of beta-catenin, a key component of the Wnt signaling pathway, in human embryonic kidney cells. Structure/function analysis reveals that complete removal of the frizzled domain of Frzb-1 abolishes the Wnt 1/Frzb-1 protein interaction and the inhibition of Wnt-1 mediated axis duplication in Xenopus embryos. In contrast, removal of the C-terminal portion of the molecule preserves both Frzb-Wnt binding and functional inhibition of Wnt signaling. Partial deletions of the Frzb-1 cysteine-rich domain maintain Wnt-1 interaction, but functional inhibition is lost. Taken together, these findings support the conclusion that the frizzled domain is necessary and sufficient for both activities. Interestingly, Frzb-1 does not block Wnt-5A signaling in a Xenopus functional assay, even though Wnt-5A coimmunoprecipitates with Frzb-1, suggesting that coimmunoprecipitation does not necessarily imply inhibition of Wnt function. PMID- 9326586 TI - Converting a transmembrane receptor to a soluble receptor: recognition domain to effector domain signaling after excision of the transmembrane domain. AB - The bacterial aspartate receptor was reconstructed to eliminate the transmembrane domain, thus connecting the recognition domain directly to the effector domain. The resulting soluble receptor folded correctly and was no longer an integral membrane protein. Upon aspartate binding, this soluble receptor was stabilized to a similar extent as that of the native receptor. Of interest, this soluble receptor retained the ability to signal from the recognition to the effector domain. This result defines more clearly the role of the membrane and transmembrane domains in signal transduction and suggests that some ligand induced motions in receptor proteins do not require the membrane or transmembrane domain for information transmission. PMID- 9326587 TI - Cockayne syndrome group B protein enhances elongation by RNA polymerase II. AB - Cockayne syndrome (CS) is characterized by impaired physical and mental development. Two complementation groups, CSA and CSB, have been identified. Here we report that the CSB gene product enhances elongation by RNA polymerase II. CSB stimulated the rate of elongation on an undamaged template by a factor of about 3. A thymine-thymine cyclobutane dimer located in the template strand is known to be a strong block to transcription. Addition of CSB to the blocked polymerase resulted in addition of one nucleotide to the nascent transcript. Finally, addition of transcription factor IIS is known to cause polymerase blocked at a thymine-thymine cyclobutane dimer to digest its nascent transcript, and CSB counteracted this transcript shortening action of transcription factor IIS. Thus a deficiency in transcription elongation may contribute to the CS phenotype. PMID- 9326588 TI - Inorganic polyphosphate and the induction of rpoS expression. AB - Inorganic polyphosphate [poly(P)] levels in Escherichia coli were reduced to barely detectable concentrations by expression of the plasmid-borne gene for a potent yeast exopolyphosphatase [poly(P)ase]. As a consequence, resistance to H2O2 was greatly diminished, particularly in katG (catalase HPI) mutants, implying a major role for the other catalase, the stationary-phase KatE (HPII), which is rpoS dependent. Resistance was restored to wild-type levels by complementation with plasmids expressing ppk, the gene for PPK [the polyphosphate kinase that generates poly(P)]. Induction of expression of both katE and rpoS (the stationary-phase sigma factor) was prevented in cells in which the poly(P)ase was overproduced. Inasmuch as this inhibition by poly(P)ase did not affect the levels of the stringent-response guanosine nucleotides (pppGpp and ppGpp) and in view of the capacity of additional rpoS expression to suppress the poly(P)ase inhibition of katE expression, a role is proposed for poly(P) in inducing the expression of rpoS. PMID- 9326589 TI - CooA, a CO-sensing transcription factor from Rhodospirillum rubrum, is a CO binding heme protein. AB - Biological sensing of small molecules such as NO, O2, and CO is an important area of research; however, little is know about how CO is sensed biologically. The photosynthetic bacterium Rhodospirillum rubrum responds to CO by activating transcription of two operons that encode a CO-oxidizing system. A protein, CooA, has been identified as necessary for this response. CooA is a member of a family of transcriptional regulators similar to the cAMP receptor protein and fumavate nitrate reduction from Escherichia coli. In this study we report the purification of wild-type CooA from its native organism, R. rubrum, to greater than 95% purity. The purified protein is active in sequence-specific DNA binding in the presence of CO, but not in the absence of CO. Gel filtration experiments reveal the protein to be a dimer in the absence of CO. Purified CooA contains 1.6 mol heme per mol of dimer. Upon interacting with CO, the electronic spectrum of CooA is perturbed, indicating the direct binding of CO to the heme of CooA. A hypothesis for the mechanism of the protein's response to CO is proposed. PMID- 9326590 TI - Recombination activities of HsDmc1 protein, the meiotic human homolog of RecA protein. AB - Meiosis-specific homologs of RecA protein have been identified in Saccharomyces cerevisiae and higher eukaryotes including mammals, but their enzymatic activities have not been described. We have purified the human protein HsDmc1 produced in Escherichia coli from a cloned copy of the cDNA. The recombinant enzyme had DNA-dependent ATPase activity with an estimated kcat of 1.5 min-1. DNase protection experiments with oligonucleotides as substrates indicated that HsDmc1 protein binds preferentially to single-stranded DNA with a stoichiometry of approximately one molecule of protein per three nucleotide residues. HsDmc1 protein catalyzed the formation of D-loops in superhelical DNA, as well as strand exchange between single-stranded and double-stranded oligonucleotides. The requirements for strand exchange catalyzed by HsDmc1 were similar to those of RecA protein, but exchange caused by HsDmc1 was not supported by ATPgammaS. PMID- 9326591 TI - Src-induced activation of inducible T cell kinase (ITK) requires phosphatidylinositol 3-kinase activity and the Pleckstrin homology domain of inducible T cell kinase. AB - The Tec family of tyrosine kinases are involved in signals emanating from cytokine receptors, antigen receptors, and other lymphoid cell surface receptors. One family member, ITK (inducible T cell kinase), is involved in T cell activation and can be activated by the T cell receptor and the CD28 cell surface receptor. This stimulation of tyrosine phosphorylation and activation of ITK can be mimicked by the Src family kinase Lck. We have explored the mechanism of this requirement for Src family kinases in the activation of ITK. We found that coexpression of ITK and Src results in increased membrane association, tyrosine phosphorylation and activation of ITK, which could be blocked by inhibitors of the lipid kinase phosphatidylinositol 3-kinase (PI 3-kinase) as well as overexpression of the p85 subunit of PI 3-kinase. Removal of the Pleckstrin homology domain (PH) of ITK resulted in a kinase that could no longer be induced to localize to the membrane or be activated by Src. The PH of ITK was also able to bind inositol phosphates phosphorylated at the D3 position. Membrane targeting of ITK without the PH recovered its ability to be activated by Src. These results suggest that ITK can be activated by a combination of Src and PI 3-kinase. PMID- 9326592 TI - Activation of protein kinase C by tyrosine phosphorylation in response to H2O2. AB - Protein kinase C (PKC) isoforms, alpha, betaI, and gamma of cPKC subgroup, delta and epsilon of nPKC subgroup, and zeta of aPKC subgroup, were tyrosine phosphorylated in COS-7 cells in response to H2O2. These isoforms isolated from the H2O2-treated cells showed enhanced enzyme activity to various extents. The enzymes, PKC alpha and delta, recovered from the cells were independent of lipid cofactors for their catalytic activity. Analysis of mutated molecules of PKC delta showed that tyrosine residues, which are conserved in the catalytic domain of the PKC family, are critical for PKC activation induced by H2O2. These results suggest that PKC isoforms can be activated through tyrosine phosphorylation in a manner unrelated to receptor-coupled hydrolysis of inositol phospholipids. PMID- 9326593 TI - Sequence-specific recognition of a subgenomic RNA promoter by a viral RNA polymerase. AB - RNA templates of 33 nucleotides containing the brome mosaic virus (BMV) core subgenomic promoter were used to determine the promoter elements recognized by the BMV RNA-dependent RNA polymerase (RdRp) to initiate RNA synthesis. Nucleotides at positions -17, -14, -13, and -11 relative to the subgenomic initiation site must be maintained for interaction with the RdRp. Changes to every other nucleotide at these four positions allow predictions for the base specific functional groups required for RdRp recognition. RdRp contact of the nucleotide at position -17 was suggested with a template competition assay. Comparison of the BMV subgenomic promoter to those from other plant and animal alphaviruses shows a remarkable degree of conservation of the nucleotides required for BMV subgenomic RNA synthesis. We show that the RdRp of the plant infecting BMV is capable of accurately, albeit inefficiently, initiating RNA synthesis from the subgenomic promoter of the animal-infecting Semliki Forest virus. The sequence-specific recognition of RNA by the BMV RdRp is analogous to the recognition of DNA promoters by DNA-dependent RNA polymerases. PMID- 9326594 TI - DNA polymerase delta isolated from Schizosaccharomyces pombe contains five subunits. AB - DNA polymerase delta (pol delta) plays an essential role in DNA replication, repair, and recombination. We have purified pol delta from Schizosaccharomyces pombe more than 10(3)-fold and demonstrated that the polymerase activity of purified S. pombe pol delta is completely dependent on proliferating cell nuclear antigen and replication factor C. SDS/PAGE analysis of the purified fraction indicated that the pol delta complex consists of five subunits that migrate with apparent molecular masses of 125, 55, 54, 42, and 22 kDa. Western blot analysis indicated that the 125, 55, and 54 kDa proteins are the large catalytic subunit (Pol3), Cdc1, and Cdc27, respectively. The identity of the other two subunits, p42 and p22, was determined following proteolytic digestion and sequence analysis of the resulting peptides. The peptide sequences derived from the p22 subunit indicated that this subunit is identical to Cdm1, previously identified as a multicopy suppressor of the temperature-sensitive cdc1-P13 mutant, whereas peptide sequences derived from the p42 subunit were identical to a previously uncharacterized ORF located on S. pombe chromosome 1. PMID- 9326595 TI - ApoNifH functions in iron-molybdenum cofactor synthesis and apodinitrogenase maturation. AB - NifH (dinitrogenase reductase) has three important roles in the nitrogenase enzyme system. In addition to its role as the obligate electron donor to dinitrogenase, NifH is required for the iron-molybdenum cofactor (FeMo-co) synthesis and apodinitrogenase maturation. We have investigated the requirement of the Fe-S cluster of NifH for these processes by preparing apoNifH. The 4Fe-4S cluster of NifH was removed by chelation of the cluster with alpha, alpha' bipyridyl. The resulting apoNifH was tested in in vitro FeMo-co synthesis and apodinitrogenase maturation reactions and was found to function in both these processes. Thus, the presence of a redox active 4Fe-4S cluster in NifH is not required for its function in FeMo-co synthesis and in apodinitrogenase maturation. This, in turn, implies that the role of NifH in these processes is not one of electron transfer or of iron or sulfur donation. PMID- 9326596 TI - The macrophage/endothelial cell mannose receptor cDNA encodes a protein that binds oligosaccharides terminating with SO4-4-GalNAcbeta1,4GlcNAcbeta or Man at independent sites. AB - Lutropin (LH) and other glycoproteins bearing oligosaccharides with the terminal sequence SO4-4-GalNAcbeta1,4GlcNAcbeta1,4Man- (S4GGnM) are rapidly removed from the circulation by an S4GGnM-specific receptor (S4GGnM-R) expressed at the surface of hepatic endothelial cells. The S4GGnM-R isolated from rat liver is closely related to the macrophage mannose-specific receptor (Man-R) isolated from rat lung both antigenically and structurally. The S4GGnM-R and Man-R isolated from these tissues nonetheless differ in their ability to bind ligands bearing terminal GalNAc-4-SO4 or Man. In this paper, we have explored the structural relationship between the Man-R and the S4GGnM-R by examining the properties of the recombinant Man-R in the form of a transmembrane protein and a soluble chimeric fusion protein in which the transmembrane and cytosolic domains have been replaced by the Fc region of human IgG1. Like the S4GGnM-R isolated from liver, the chimeric fusion protein is able to bind ligands terminating with GalNAc-4-SO4 and Man at independent sites. When expressed in CHO cells the recombinant Man-R is able to mediate the uptake of ligands bearing either terminal GalNAc-4-SO4 or terminal Man. We propose that the Man-R be renamed the Man/S4GGnM receptor on the basis of its multiple and independent specificities. PMID- 9326597 TI - Chrysanthemum chlorotic mottle viroid: unusual structural properties of a subgroup of self-cleaving viroids with hammerhead ribozymes. AB - The causal agent of chrysanthemum chlorotic mottle (CChM) disease has been identified, cloned, and sequenced. It is a viroid RNA (CChMVd) of 398-399 nucleotides. In vitro transcripts with the complete CChMVd sequence were infectious and induced the typical symptoms of the CChM disease. CChMVd can form hammerhead structures in both polarity strands. Plus and minus monomeric CChMVd RNAs self-cleaved during in vitro transcription and after purification as predicted by these structures, which are stable and most probably act as single hammerhead structures as in peach latent mosaic viroid (PLMVd), but not in avocado sunblotch viroid (ASBVd). Moreover, the plus CChMVd hammerhead structure also appears to be active in vivo, because the 5' terminus of the linear plus CChMVd RNA isolated from infected tissue is that predicted by the corresponding hammerhead ribozyme. Both hammerhead structures of CChMVd display some peculiarities: the plus self-cleaving domain has an unpaired A after the conserved A9 residue, and the minus one has an unusually long helix II. The most stable secondary structure predicted for CChMVd is a branched conformation that does not fulfill the rod-like or quasi-rod-like model proposed for the in vitro structure of most viroids with the exception of PLMVd, whose proposed secondary structure of lowest free energy also is branched. The unusual conformation of CChMVd and PLMVd is supported by their insolubility in 2 M LiCl, in contrast to ASBVd and a series of representative non-self-cleaving viroids that are soluble under the same high salt conditions. These results support the classification of self-cleaving viroids into two subgroups, one formed by ASBVd and the other one by PLMVd and CChMVd. PMID- 9326599 TI - Positioning of two alpha subunit carboxy-terminal domains of RNA polymerase at promoters by two transcription factors. AB - Interactions between the cAMP receptor protein (CRP) and the carboxy-terminal regulatory domain (CTD) of Escherichia coli RNA polymerase alpha subunit were analyzed at promoters carrying tandem DNA sites for CRP binding using a chemical nuclease covalently attached to alpha. Each CRP dimer was found to direct the positioning of one of the two alpha subunit CTDs. Thus, the function of RNA polymerase may be subject to regulation through protein-protein interactions between the two alpha subunits and two different species of transcription factors. PMID- 9326598 TI - RB and hbrm cooperate to repress the activation functions of E2F1. AB - Forced expression of the retinoblastoma (RB) gene product inhibits the proliferation of cells in culture. A major target of the RB protein is the S phase-inducing transcription factor E2F1. RB binds directly to the activation domain of E2F1 and silences it, thereby preventing cells from entering S phase. To induce complete G1 arrest, RB requires the presence of the hbrm/BRG-1 proteins, which are components of the coactivator SWI/SNF complex. This cooperation is mediated through a physical interaction between RB and hbrm/BRG-1. We show here that in transfected cells RB can contact both E2F1 and hbrm at the same time, thereby targeting hbrm to E2F1. E2F1 and hbrm are indeed found within the same complex in vivo. Furthermore, RB and hbrm cooperate to repress E2F1 activity in transient transfection assays. The ability of hbrm to cooperate with RB to repress E2F1 is dependent upon several distinct domains of hbrm, including the RB binding domain and the NTP binding site. However, the bromodomain seems dispensable for this activity. Taken together, our results point out an unexpected role of corepressor for the hbrm protein. The ability of hbrm and RB to cooperate in repressing E2F1 activity could be an underlying mechanism for the observed cooperation between hbrm and RB to induce G1 arrest. Finally, we demonstrate that the domain of hbrm that binds RB has transcriptional activation potential which RB can repress. This suggest that RB not only targets hbrm but also regulates its activity. PMID- 9326600 TI - Binding of RNA template to a complex of HIV-1 reverse transcriptase/primer/template. AB - HIV-1 reverse transcriptase (RT) catalyzes the synthesis of DNA from DNA or RNA templates. During this process, it must transfer its primer from one template to another RNA or DNA template. Binary complexes made of RT and a primer/template bind an additional single-stranded RNA molecule of the same nucleotide sequence as that of the DNA or RNA template. The additional RNA strand leads to a 10-fold decrease of the off-rate constant, koff, of RT from a primer/DNA template. In a binary complex of RT and a primer/template, the primer can be cross-linked to both the p66 and p51 subunits. Depending on the location of the photoreactive group in the primer, the distribution of the cross-linked primers between subunits is dependent on the nature of the template and of the additional single stranded molecule. Greater cross-linking of the primer to p51 occurs with DNA templates, whereas cross-linking to p66 predominates with RNA templates. Excess single-stranded DNA shifts the distribution of cross-linking from p66 to p51 with RNA templates, and excess single-stranded RNA shifts the cross-linking from p51 to p66 with DNA templates. RT thus uses two primer/template binding modes depending on the nature of the template. PMID- 9326601 TI - Bioactive and nuclease-resistant L-DNA ligand of vasopressin. AB - In vitro selection experiments have produced nucleic acid ligands (aptamers) that bind tightly and specifically to a great variety of target biomolecules. The utility of aptamers is often limited by their vulnerability to nucleases present in biological materials. One way to circumvent this problem is to select an aptamer that binds the enantiomer of the target, then synthesize the enantiomer of the aptamer as a nuclease-insensitive ligand of the normal target. We have so identified a mirror-image single-stranded DNA that binds the peptide hormone vasopressin and have demonstrated its stability to nucleases and its bioactivity as a vasopressin antagonist in cell culture. PMID- 9326602 TI - Subtle atomic group discrimination in the RNA minor groove. AB - As a problem in molecular recognition and for drug discovery, great interest has developed around the possibility that RNA structures could be discriminated by peptides and other small molecules. Although small peptides have been shown to have the capacity to discriminate specific bulges and loops in RNA molecules, discrimination of double helical regions by a peptide binder has not been reported. Indeed, the most accessible part of an RNA helix is the minor groove, and fundamental stereochemical considerations have suggested that discrimination of at least some base pairs would be difficult in the minor groove. Here we report the design and isolation of a peptide binder that manifests the most subtle kind of discrimination of base pair differences in the RNA minor groove. Functional discrimination of a single atomic group is demonstrated as well as the difference between two different angular orientations of the same group. This report of RNA helix discrimination by a peptide binder suggests a richer potential for RNA minor groove recognition than previously thought. PMID- 9326603 TI - A histone deacetylase inhibitor potentiates retinoid receptor action in embryonal carcinoma cells. AB - Histone acetylation is thought to have a role in transcription. To gain insight into the role of histone acetylation in retinoid-dependent transcription, we studied the effects of trichostatin A (TSA), a specific inhibitor of histone deacetylase, on P19 embryonal carcinoma cells. We show that coaddition of TSA and retinoic acid (RA) markedly enhances neuronal differentiation in these cells, although TSA alone does not induce differentiation but causes extensive apoptosis. Consistent with the cooperative effect of TSA and RA, coaddition of the two agents synergistically enhanced transcription from stably integrated RA responsive promoters. The transcriptional synergy by TSA and RA required the RA responsive element and a functional retinoid X receptor (RXR)/retinoic acid receptor (RAR) heterodimer, both obligatory for RA-dependent transcription. Furthermore, TSA led to promoter activation by an RXR-selective ligand that was otherwise inactive in transcription. In addition, TSA enhanced transcription from a minimum basal promoter, independently of the RA-responsive element. Finally, we show that TSA alone or in combination with RA increases in vivo endonuclease sensitivity within the RA-responsive promoter, suggesting that TSA treatment might alter a local chromatin environment to enhance RXR/RAR heterodimer action. Thus, these results indicate that histone acetylation influences activity of the heterodimer, which is in line with the observed interaction between the RXR/RAR heterodimer and a histone acetylase presented elsewhere. PMID- 9326605 TI - The efficiency of propulsion by a rotating flagellum. AB - At very low Reynolds number, the regime in which fluid dynamics is governed by Stokes equations, a helix that translates along its axis under an external force but without an external torque will necessarily rotate. By the linearity of the Stokes equations, the same helix that is caused to rotate due to an external torque will necessarily translate. This is the physics that underlies the mechanism of flagellar propulsion employed by many microorganisms. Here, I examine the linear relationships between forces and torques and translational and angular velocities of helical objects to understand the nature of flagellar propulsion. PMID- 9326604 TI - A functionally defined model for the M2 proton channel of influenza A virus suggests a mechanism for its ion selectivity. AB - The M2 protein from influenza A virus forms proton-selective channels that are essential to viral function and are the target of the drug amantadine. Cys scanning was used to generate a series of mutants with successive substitutions in the transmembrane segment of the protein, and the mutants were expressed in Xenopus laevis oocytes. The effect of the mutations on reversal potential, ion currents, and amantadine resistance were measured. Fourier analysis revealed a periodicity consistent with a four-stranded coiled coil or helical bundle. A three-dimensional model of this structure suggests a possible mechanism for the proton selectivity of the M2 channel of influenza virus. PMID- 9326606 TI - Raman spectral evidence for hydration forces between collagen triple helices. AB - Hydration forces are thought to result from the energetic cost of water rearrangement near macromolecular surfaces. Raman spectra, collected on the same collagen samples on which these forces were measured, reveal a continuous change in water hydrogen-bonding structure as a function of separation between collagen triple helices. The varying spectral parameters track the force-distance curve. The energetic cost of water "restructuring," estimated from the spectra, is consistent with the measured energy of intermolecular interaction. These correlations support the idea that the change in water structure underlies the exponentially varying forces seen in this system at least over the 13-18-A range of interaxial separations. PMID- 9326608 TI - Evolution of the folding ability of proteins through functional selection. AB - An evolutionary process is simulated with a simple spin-glass-like model of proteins to examine the origin of folding ability. At each generation, sequences are randomly mutated and subjected to a simulation of the folding process based on the model. According to the frequency of local configurations at the active sites, sequences are selected and passed to the next generation. After a few hundred generations, a sequence capable of folding globally into a native conformation emerges. Moreover, the selected sequence has a distinct energy minimum and an anisotropic funnel on the energy surface, which are the imperative features for fast folding of proteins. The proposed model reveals that the functional selection on the local configurations leads a sequence to fold globally into a conformation at a faster rate. PMID- 9326609 TI - Stability and dynamics in a hyperthermophilic protein with melting temperature close to 200 degrees C. AB - The rubredoxin protein from the hyperthermophilic archaebacterium Pyrococcus furiosus was examined by a hydrogen exchange method. Even though the protein does not exhibit reversible thermal unfolding, one can determine its stability parameters-free energy, enthalpy, entropy, and melting temperature-and also the distribution of stability throughout the protein, by using hydrogen exchange to measure the reversible cycling of the protein between native and unfolded states that occurs even under native conditions. PMID- 9326610 TI - MRIT, a novel death-effector domain-containing protein, interacts with caspases and BclXL and initiates cell death. AB - Activation of the cascade of proteolytic caspases has been identified as the final common pathway of apoptosis in diverse biological systems. We have isolated a gene, termed MRIT, that possesses overall sequence homology to FLICE (MACH), a large prodomain caspase that links the aggregated complex of the death domain receptors of the tumor necrosis factor receptor family to downstream caspases. However, unlike FLICE, the C-terminal domain of MRIT lacks the caspase catalytic consensus sequence QAC(R/Q)G. Nonetheless MRIT activates caspase-dependent death. Using yeast two-hybrid assays, we demonstrate that MRIT associates with caspases possessing large and small prodomains (FLICE, and CPP32/YAMA), as well as with the adaptor molecule FADD. In addition, MRIT simultaneously and independently interacts with BclXL and FLICE in mammalian cells. Thus, MRIT is a mammalian protein that interacts simultaneously with both caspases and a Bcl-2 family member. PMID- 9326611 TI - The signal recognition particle receptor of Escherichia coli (FtsY) has a nucleotide exchange factor built into the GTPase domain. AB - Targeting of many secretory and membrane proteins to the inner membrane in Escherichia coli is achieved by the signal recognition particle (SRP) and its receptor (FtsY). In E. coli SRP consists of only one polypeptide (Ffh), and a 4.5S RNA. Ffh and FtsY each contain a conserved GTPase domain (G domain) with an alpha-helical domain on its N terminus (N domain). The nucleotide binding kinetics of the NG domain of the SRP receptor FtsY have been investigated, using different fluorescence techniques. Methods to describe the reaction kinetically are presented. The kinetics of interaction of FtsY with guanine nucleotides are quantitatively different from those of other GTPases. The intrinsic guanine nucleotide dissociation rates of FtsY are about 10(5) times higher than in Ras, but similar to those seen in GTPases in the presence of an exchange factor. Therefore, the data presented here show that the NG domain of FtsY resembles a GTPase-nucleotide exchange factor complex not only in its structure but also kinetically. The I-box, an insertion present in all SRP-type GTPases, is likely to act as an intrinsic exchange factor. From this we conclude that the details of the GTPase cycle of FtsY and presumably other SRP-type GTPases are fundamentally different from those of other GTPases. PMID- 9326612 TI - Interleukin 3-dependent survival by the Akt protein kinase. AB - Interleukin 3 (IL-3)-dependent survival of hematopoietic cells is known to rely on the activity of multiple signaling pathways, including a pathway leading to activation of phosphoinositide 3-kinase (PI 3-kinase), and protein kinase Akt is a direct target of PI 3-kinase. We find that Akt kinase activity is rapidly induced by the cytokine IL-3, suggesting a role for Akt in PI 3-kinase-dependent signaling in hematopoetic cells. Dominant-negative mutants of Akt specifically block Akt activation by IL-3 and interfere with IL-3-dependent proliferation. Overexpression of Akt or oncogenic v-akt protects 32D cells from apoptosis induced by IL-3 withdrawal. Apoptosis after IL-3 withdrawal is accelerated by expression of dominant-negative mutants of Akt, indicating that a functional Akt signaling pathway is necessary for cell survival mediated by the cytokine IL-3. Thus Akt appears to be an important mediator of anti-apoptotic signaling in this system. PMID- 9326613 TI - A transformation-associated complex involving tyrosine kinase signal adapter proteins and caldesmon links v-erbB signaling to actin stress fiber disassembly. AB - The avian erythroblastosis viral oncogene (v-erbB) encodes a receptor tyrosine kinase that possesses sarcomagenic and leukemogenic potential. We have expressed transforming and nontransforming mutants of v-erbB in fibroblasts to detect transformation-associated signal transduction events. Coimmunoprecipitation and affinity chromatography have been used to identify a transformation-associated, tyrosine phosphorylated, multiprotein complex. This complex consists of Src homologous collagen protein (Shc), growth factor receptor binding protein 2 (Grb2), son of sevenless (Sos), and a novel tyrosine phosphorylated form of the cytoskeletal regulatory protein caldesmon. Immunofluorescence localization studies further reveal that, in contrast to the distribution of caldesmon along actin stress fibers in normal fibroblasts, caldesmon colocalizes with Shc in plasma membrane blebs in transformed fibroblasts. This colocalization of caldesmon and Shc correlates with actin stress fiber disassembly and v-erbB mediated transformation. The tyrosine phosphorylation of caldesmon, and its association with the Shc-Grb2-Sos signaling complex directly links tyrosine kinase oncogenic signaling events with cytoskeletal regulatory processes, and may define one mechanism regulating actin stress fiber disassembly in transformed cells. PMID- 9326614 TI - Comparison of the ion channel characteristics of proapoptotic BAX and antiapoptotic BCL-2. AB - The BCL-2 family of proteins is composed of both pro- and antiapoptotic regulators, although its most critical biochemical functions remain uncertain. The structural similarity between the BCL-XL monomer and several ion-pore-forming bacterial toxins has prompted electrophysiologic studies. Both BAX and BCL-2 insert into KCl-loaded vesicles in a pH-dependent fashion and demonstrate macroscopic ion efflux. Release is maximum at approximately pH 4.0 for both proteins; however, BAX demonstrates a broader pH range of activity. Both purified proteins also insert into planar lipid bilayers at pH 4.0. Single-channel recordings revealed a minimal channel conductance for BAX of 22 pS that evolved to channel currents with at least three subconductance levels. The final, apparently stable BAX channel had a conductance of 0.731 nS at pH 4. 0 that changed to 0.329 nS when shifted to pH 7.0 but remained mildly Cl- selective and predominantly open. When BAX-incorporated lipid vesicles were fused to planar lipid bilayers at pH 7.0, a Cl--selective (PK/PCl = 0.3) 1.5-nS channel displaying mild inward rectification was noted. In contrast, BCL-2 formed mildly K+-selective (PK/PCl = 3.9) channels with a most prominent initial conductance of 80 pS that increased to 1.90 nS. Fusion of BCL-2-incorporated lipid vesicles into planar bilayers at pH 7.0 also revealed mild K+ selectivity (PK/PCl = 2.4) with a maximum conductance of 1.08 nS. BAX and BCL-2 each form channels in artificial membranes that have distinct characteristics including ion selectivity, conductance, voltage dependence, and rectification. Thus, one role of these molecules may include pore activity at selected membrane sites. PMID- 9326615 TI - Endoplasmic reticulum quality control of asialoglycoprotein receptor H2a involves a determinant for retention and not retrieval. AB - The human asialoglycoprotein receptor H2a subunit contains a charged pentapeptide, EGHRG, in its ectodomain that is the only sequence absent from the H2b alternatively spliced variant. H2b exits the endoplasmic reticulum (ER) even when singly expressed, whereas H2a gives rise to a cleaved soluble secreted ectodomain fragment; uncleaved membrane-bound H2a molecules are completely retained and degraded in the ER. We have inserted the H2a pentapeptide into the sequence of the H1 subunit (H1i5), which caused complete ER retention but, unexpectedly, no degradation. This suggests that the pentapeptide is a determinant for ER retention not colocalizing in H2a with the determinant for degradation. The state of sugar chain processing and the ER localization of H1i5, which was unchanged at 15 degrees C or after treatment with nocodazole, indicate ER retention and not retrieval from the cis-Golgi or the intermediate compartment. H1i5 folded similarly to H1, and both associated to calnexin. However, whereas H1 dissociated with a half time of 45 min, H1i5 remained bound to the chaperone for prolonged periods. The correct global folding of H2a and H1i5 and of other normal precursors and unassembled proteins and the true ER retention, and not exit and retrieval, suggest a difference in their quality control mechanism compared with that of misfolded proteins, which does involve retrieval. However, both pathways may involve calnexin. PMID- 9326616 TI - Integration of multiple instructive cues by neural crest stem cells reveals cell intrinsic biases in relative growth factor responsiveness. AB - Growth factors can influence lineage determination of neural crest stem cells (NCSCs) in an instructive manner, in vitro. Because NCSCs are likely exposed to multiple signals in vivo, these findings raise the question of how stem cells would integrate such combined influences. Bone morphogenetic protein 2 (BMP2) promotes neuronal differentiation and glial growth factor 2 (GGF2) promotes glial differentiation; if NCSCs are exposed to saturating concentrations of both factors, BMP2 appears dominant. By contrast, if the cells are exposed to saturating concentrations of both BMP2 and transforming growth factor beta1 (which promotes smooth muscle differentiation), the two factors appear codominant. Sequential addition experiments indicate that NCSCs require 48-96 hrs in GGF2 before they commit to a glial fate, whereas the cells commit to a smooth muscle fate within 24 hr in transforming growth factor beta1. The delayed response to GGF2 does not reflect a lack of functional receptors; however, because the growth factor induces rapid mitogen-activated protein kinase phosphorylation in naive cells. Furthermore, GGF2 can attenuate induction of the neurogenic transcription factor mammalian achaete-scute homolog 1, by low doses of BMP2. This short-term antineurogenic influence of GGF2 is not sufficient for glial lineage commitment, however. These data imply that NCSCs exhibit cell intrinsic biases in the timing and relative dosage sensitivity of their responses to instructive factors that influence the outcome of lineage decisions in the presence of multiple factors. The relative delay in glial lineage commitment, moreover, apparently reflects successive short-term and longer-term actions of GGF2. Such a delay may help to explain why glia normally differentiate after neurons, in vivo. PMID- 9326617 TI - Role of lipid polymorphism in G protein-membrane interactions: nonlamellar-prone phospholipids and peripheral protein binding to membranes. AB - Heterotrimeric G proteins (peripheral proteins) conduct signals from membrane receptors (integral proteins) to regulatory proteins localized to various cellular compartments. They are in excess over any G protein-coupled receptor type on the cell membrane, which is necessary for signal amplification. These facts account for the large number of G protein molecules bound to membrane lipids. Thus, the protein-lipid interactions are crucial for their cellular localization, and consequently for signal transduction. In this work, the binding of G protein subunits to model membranes (liposomes), formed with defined membrane lipids, has been studied. It is shown that although G protein alpha subunits were able to bind to lipid bilayers, the presence of nonlamellar-prone phospholipids (phosphatidylethanolamines) enhanced their binding to model membranes. This mechanism also appears to be used by other (structurally and functionally unrelated) peripheral proteins, such as protein kinase C and the insect protein apolipophorin III, indicating that it could constitute a general mode of protein-lipid interactions, relevant in the activity and translocation of some peripheral (amphitropic) proteins from soluble to particulate compartments. Other factors, such as the presence of cholesterol or the vesicle surface charge, also modulated the binding of the G protein subunits to lipid bilayers. Conversely, the binding of G protein-coupled receptor kinase 2 and the G protein beta-subunit to liposomes was not increased by hexagonally prone lipids. Their distinct interactions with membrane lipids may, in part, explain the different cellular localizations of all of these proteins during the signaling process. PMID- 9326618 TI - Insulin and insulin-like growth factor-I acutely inhibit surface translocation of growth hormone receptors in osteoblasts: a novel mechanism of growth hormone receptor regulation. AB - We previously have demonstrated that insulin and insulin-like growth factor-I (IGF-I) down-regulate growth hormone (GH) binding in osteoblasts by reducing the number of surface GH receptors (GHRs). The present study was undertaken to investigate the mechanism of GHR down-regulation. Treatment with 5 nM insulin or IGF-I for 18 hr significantly decreased surface GH binding to 26.4 +/- 2.9% and 23.0 +/- 2.7% of control (mean +/- SE; P < 0.05), respectively. No corresponding reductions in the mRNA level and total cellular content of GHR were found, nor was the rate of receptor internalization affected. The effects on GHR translocation were assessed by measuring the reappearance of GH binding of whole cells after trypsinization to remove the surface receptors. GH binding of control cultures significantly increased (P < 0.05) over 2 hr after trypsinization, whereas no recovery of binding activity was detected in insulin and IGF-I-treated cultures, indicating that GHR translocation was impaired. Studies on the time course of GHR down-regulation revealed that surface GH binding was reduced significantly by 3-hr treatment (P /=50 kb) and nuclear fragmentation. Important to note, specific inhibition of PAO by N,N'-bis(2, 3-butadienyl)-1,4-butane-diamine results in a significant reduction of the formation of HMW DNA and nuclear fragmentation. In contrast, the coaddition of catalase or PAO inhibitor has no effect on reducing HMW DNA fragmentation induced by N1-ethyl-N11-[(cycloheptyl)methyl]-4,8,-diazaundecane, which does not induce SSAT and does not deplete intracellular polyamines. These results strongly suggest that H2O2 production by PAO has a role in CPENSpm cytotoxicity in sensitive cells via PCD and demonstrate a potential basis for differential sensitivity to this promising new class of antineoplastic agents. Furthermore, the data suggest a general mechanism by which, under certain stimuli, cells can commit suicide through catabolism of the ubiquitous intracellular polyamines. PMID- 9326649 TI - A factor IX-deficient mouse model for hemophilia B gene therapy. AB - We have generated a mouse where the clotting factor IX (FIX) gene has been disrupted by homologous recombination. The FIX nullizygous (-/-) mouse was devoid of factor IX antigen in plasma. Consistent with the bleeding disorder, the factor IX coagulant activities for wild-type (+/+), heterozygous (+/-), and homozygous ( /-) mice were 92%, 53%, and <5%, respectively, in activated partial thromboplastin time assays. Plasma factor IX activity in the deficient mice (-/-) was restored by introducing wild-type murine FIX gene via adenoviral vectors. Thus, these factor IX-deficient mice provide a useful animal model for gene therapy studies of hemophilia B. PMID- 9326650 TI - Phenotypic knockout of HIV type 1 chemokine coreceptor CCR-5 by intrakines as potential therapeutic approach for HIV-1 infection. AB - A genetic defect in a CC-chemokine receptor (CCR)-5, the principal coreceptor for the macrophage-tropic HIV type 1 (HIV-1), recently was found to naturally protect CCR-5-defective, but healthy, individuals from HIV-1 infection. In this study, we mimic the natural resistance of the CCR-5-defective individuals by designing a strategy to phenotypically knock out CCR-5. The inactivation of the CCR-5 coreceptor is accomplished by targeting a modified CC-chemokine to the endoplasmic reticulum to block the surface expression of newly synthesized CCR-5. The lymphocytes transduced to express the intracellular chemokine, termed "intrakine," were found to be viable and resistant to macrophage-tropic HIV-1 infection. Thus, this gene-based intrakine strategy targeted at the conserved cellular receptor for the prevention of HIV-1 entry should have significant advantages over currently described approaches for HIV-1 therapy. PMID- 9326651 TI - Governing step of metastasis visualized in vitro. AB - Metastasis is the ultimate life-threatening stage of cancer. The lack of accurate model systems thwarted studies of the metastatic cell's basic biology. To follow continuously the succeeding stages of metastatic colony growth, we heritably labeled cells from the human lung adenocarcinoma cell line ANIP 973 with green fluorescent protein (GFP) by transfection with GFP cDNA. Labeled cells were then injected intravenously into nude mice, where, by 7 days, they formed brilliantly fluorescing metastatic colonies on mouse lung [Chishima, T., Miyagi, Y., Wang, X., Yang, M., Tan, Y., Shimada, H., Moossa, A. R. & Hoffman, R. M. (1997) Clin. Exp. Metastasis 15, 547-552]. The seeded lung tissue was then excised and incubated in the three-dimensional sponge-gel-matrix-supported histoculture that maintained the critical features of progressive in vivo tumor colonization while allowing continuous access for measurement and manipulation. Tumor progression was continuously visualized by GFP fluorescence in the same individual cultures over a 52-day period, during which the tumors spread throughout the lung. Histoculture tumor colonization was selective for lung cancer cells to grow on lung tissue, because no growth occurred on histocultured mouse liver tissue, which was also observed in vivo. The ability to support selective organ colonization in histoculture and visualize tumor progression by GFP fluorescence allows the in vitro study of the governing processes of metastasis [Kuo, T.-H., Kubota, T., Watanbe, M., Furukawa, T., Teramoto, T., Ishibiki, K., Kitajima, M., Moossa, A. R., Penman, S. & Hoffman, R. M. (1995) Proc. Natl. Acad. Sci. USA 92, 12085-12089]. The results presented here provide significant, new opportunities to understand and to develop treatments that prevent and possibly reverse metastasis. PMID- 9326652 TI - Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes (PMN) in the presence of EDTA: PMN in close quarters require neither leukocyte integrins nor external divalent cations. AB - Divalent cations are thought essential for motile function of leukocytes in general, and for the function of critical adhesion molecules in particular. In the current study, under direct microscopic observation with concomitant time lapse video recording, we examined the effects of 10 mM EDTA on locomotion of human blood polymorphonuclear leukocytes (PMN). In very thin slide preparations, EDTA did not impair either random locomotion or chemotaxis; motile behavior appeared to benefit from the close approximation of slide and coverslip ("chimneying"). In preparations twice as thick, PMN in EDTA first exhibited active deformability with little or no displacement, then rounded up and became motionless. However, on creation of a chemotactic gradient, the same cells were able to orient and make their way to the target, often, however, losing momentarily their purchase on the substrate. In either of these preparations without EDTA, specific antibodies to beta2 integrins did not prevent random locomotion or chemotaxis, even when we added antibodies to beta1 and alphavbeta3 integrins and to integrin-associated protein, and none of these antibodies added anything to the effects of EDTA. In the more turbulent environment of even more media, effects of anti-beta2 integrins became evident: PMN still could locomote but adhered to substrate largely by their uropods and by uropod-associated filaments. We relate these findings to the reported independence from integrins of PMN in certain experimental and disease states. Moreover, we suggest that PMN locomotion in close quarters is not only integrin-independent, but independent of external divalent cations as well. PMID- 9326653 TI - Panhandle PCR strategy to amplify MLL genomic breakpoints in treatment-related leukemias. AB - Panhandle PCR amplifies genomic DNA with known 5' and unknown 3' sequences from a template with an intrastrand loop schematically shaped like a pan with a handle. We used panhandle PCR to clone MLL genomic breakpoints in two pediatric treatment related leukemias. The karyotype in a case of treatment-related acute lymphoblastic leukemia showed the t(4;11)(q21;q23). Panhandle PCR amplified the translocation breakpoint at position 2158 in intron 6 in the 5' MLL breakpoint cluster region (bcr). The karyotype in a case of treatment-related acute myeloid leukemia was normal, but Southern blot analysis showed a single MLL gene rearrangement. Panhandle PCR amplified the breakpoint at position 1493 in MLL intron 6. Screening of somatic cell hybrid and radiation hybrid DNAs by PCR and reverse transcriptase-PCR analysis of the leukemic cells indicated that panhandle PCR identified a fusion of MLL intron 6 with a previously uncharacterized sequence in MLL intron 1, consistent with a partial duplication. In both cases, the breakpoints in the MLL bcr were in Alu repeats, and there were Alu repeats in proximity to the breakpoints in the partner DNAs, suggesting that Alu sequences were relevant to these rearrangements. This study shows that panhandle PCR is an effective method for cloning MLL genomic breakpoints in treatment-related leukemias. Analysis of additional pediatric cases will determine whether breakpoint distribution deviates from the predilection for 3' distribution in the bcr that has been found in adult cases. PMID- 9326655 TI - Interferon beta transduction of peripheral blood lymphocytes from HIV-infected donors increases Th1-type cytokine production and improves the proliferative response to recall antigens. AB - We are developing a gene therapy method of HIV infection based on the constitutive low production of interferon (IFN) beta. Peripheral blood lymphocytes (PBL) from HIV-infected patients at different clinical stages of infection were efficiently transduced with the HMB-HbHuIFNbeta retroviral vector. The constitutive low production of IFN-beta in cultured PBL from HIV-infected patients resulted in a decreased viral production and an enhanced survival of CD4+ cells, and this protective effect was observed only in the PBL derived from donors having a CD4+ cell count above 200 per mm3. In IFN-beta-transduced PBL from healthy and from HIV-infected donors, the production of the Th1-type cytokines IFN-gamma and interleukin (IL)-12 was enhanced. In IFN-beta-transduced PBL from HIV-infected donors, the production of IL-4, IL-6, IL-10, and tumor necrosis factor alpha was maintained at normal levels, contrary to the increased levels produced by the untransduced PBL. The proliferative response to recall antigens was partially restored in IFN-beta-transduced PBL from donors with an impaired antigen response. Thus, in addition to inhibiting HIV replication, IFN beta transduction of PBL from HIV-infected donors improves several parameters of immune function. PMID- 9326654 TI - Decreased blood pressure response in mice deficient of the alpha1b-adrenergic receptor. AB - To investigate the functional role of different alpha1-adrenergic receptor (alpha1-AR) subtypes in vivo, we have applied a gene targeting approach to create a mouse model lacking the alpha1b-AR (alpha1b-/-). Reverse transcription-PCR and ligand binding studies were combined to elucidate the expression of the alpha1-AR subtypes in various tissues of alpha1b +/+ and -/- mice. Total alpha1-AR sites were decreased by 98% in liver, 74% in heart, and 42% in cerebral cortex of the alpha1b -/- as compared with +/+ mice. Because of the large decrease of alpha1-AR in the heart and the loss of the alpha1b-AR mRNA in the aorta of the alpha1b-/- mice, the in vivo blood pressure and in vitro aorta contractile responses to alpha1-agonists were investigated in alpha1b +/+ and -/- mice. Our findings provide strong evidence that the alpha1b-AR is a mediator of the blood pressure and the aorta contractile responses induced by alpha1 agonists. This was demonstrated by the finding that the mean arterial blood pressure response to phenylephrine was decreased by 45% in alpha1b -/- as compared with +/+ mice. In addition, phenylephrine-induced contractions of aortic rings also were decreased by 25% in alpha1b-/- mice. The alpha1b-AR knockout mouse model provides a potentially useful tool to elucidate the functional specificity of different alpha1-AR subtypes, to better understand the effects of adrenergic drugs, and to investigate the multiple mechanisms involved in the control of blood pressure. PMID- 9326656 TI - Developmental disorder associated with increased cellular nucleotidase activity. AB - Four unrelated patients are described with a syndrome that included developmental delay, seizures, ataxia, recurrent infections, severe language deficit, and an unusual behavioral phenotype characterized by hyperactivity, short attention span, and poor social interaction. These manifestations appeared within the first few years of life. Each patient displayed abnormalities on EEG. No unusual metabolites were found in plasma or urine, and metabolic testing was normal except for persistent hypouricosuria. Investigation of purine and pyrimidine metabolism in cultured fibroblasts derived from these patients showed normal incorporation of purine bases into nucleotides but decreased incorporation of uridine. De novo synthesis of purines and cellular phosphoribosyl pyrophosphate content also were moderately decreased. The distribution of incorporated purines and pyrimidines did not reveal a pattern suggestive of a deficient enzyme activity. Assay of individual enzymes in fibroblast lysates showed no deficiencies. However, the activity of cytosolic 5'-nucleotidase was elevated 6- to 10-fold. Based on the possibility that the observed increased catabolic activity and decreased pyrimidine salvage might be causing a deficiency of pyrimidine nucleotides, the patients were treated with oral pyrimidine nucleoside or nucleotide compounds. All patients showed remarkable improvement in speech and behavior as well as decreased seizure activity and frequency of infections. A double-blind placebo trial was undertaken to ascertain the efficacy of this supplementation regimen. Upon replacement of the supplements with placebo, all patients showed rapid regression to their pretreatment states. These observations suggest that increased nucleotide catabolism is related to the symptoms of these patients, and that the effects of this increased catabolism are reversed by administration of uridine. PMID- 9326657 TI - Transmembrane beta-barrel of staphylococcal alpha-toxin forms in sensitive but not in resistant cells. AB - Staphylococcal alpha-toxin is a 293-residue, single-chain polypeptide that spontaneously assembles into a heptameric pore in target cell membranes. To identify the pore-forming domain, substitution mutants have been produced in which single cysteine residues were introduced throughout the toxin molecule. By attaching the environmentally sensitive dye acrylodan to the sulfhydryl groups, the environment of individual amino acid side chains could be probed. In liposomes, a single 23-amino acid sequence (residues 118-140) was found to move from a polar to a nonpolar environment, indicating that this sequence forms the walls of the pore. However, periodicity in side chain environmental polarity could not be detected in the liposomal system. In the present study, the fluorimetric analyses were extended to physiological target cells. With susceptible cells such as rabbit erythrocytes and human lymphocytes, the 23 central amino acids 118-140 were again found to insert into the membrane; in contrast to the previous study with liposomes, the expected periodicity was now detected. Thus, every other residue in the sequence 126-140 entered a nonpolar environment in a striking display of an amphipathic transmembrane beta-barrel. In contrast, human granulocytes were found to bind alpha-toxin to a similar extent as lymphocytes, but the heptamers forming on these cells failed to insert their pore-forming domain into the membrane. As a consequence, nonfunctional heptamers assembled and the cells remained viable. The data resolve the molecular organization of a pore-forming toxin domain in living cells and reveal that resistant cells can prevent insertion of the functional domain into the bilayer. PMID- 9326658 TI - Binding of high-risk human papillomavirus E6 oncoproteins to the human homologue of the Drosophila discs large tumor suppressor protein. AB - In the majority of cervical cancers, DNAs of high-risk mucosotpropic human papillomaviruses (HPVs), such as type 16, are maintained so as to express two viral proteins, E6 and E7, suggesting an essential importance to carcinogenesis. The high-risk HPV E6 proteins are known to inactivate p53 tumor suppressor protein but appear to have an additional, molecularly unknown function(s). In this study, we demonstrate that these E6 proteins can bind to the second PDZ domain of the human homologue of the Drosophila discs large tumor suppressor protein (hDLG) through their C-terminal XS/TXV/L (where X represents any amino acid, S/T serine or threonine, and V/L valine or leucine) motif. This finding is similar to the interaction between the adenomatous polyposis coli gene product and hDLG. E6 mutants losing the ability to bind to hDLG are no longer able to induce E6-dependent transformation of rodent cells. These results suggest an intriguing possibility that interaction between the E6 protein and hDLG or other PDZ domain-containing proteins could be an underlying mechanism in the development of HPV-associated cancers. PMID- 9326659 TI - Identification of a cellular receptor for subgroup E avian leukosis virus. AB - Genetic studies in chickens and receptor interference experiments have indicated that avian leukosis virus (ALV)-E may utilize a cellular receptor related to the receptor for ALV-B and ALV-D. Recently, we cloned CAR1, a tumor necrosis factor receptor (TNFR)-related protein, that serves as a cellular receptor for ALV-B and ALV-D. To determine whether the cellular receptor for ALV-E is a CAR1-like protein, a cDNA library was made from turkey embryo fibroblasts (TEFs), which are susceptible to ALV-E infection, but not to infection by ALV-B and ALV-D. The cDNA library was screened with a radioactively labeled CAR1 cDNA probe, and clones that hybridized with the probe were isolated. A 2.3-kb cDNA clone was identified that conferred susceptibility to ALV-E infection, but not to ALV-B infection, when expressed in transfected human 293 cells. The functional cDNA clone is predicted to encode a 368 amino acid protein with significant amino acid similarity to CAR1. Like CAR1, the TEF protein is predicted to have two extracellular TNFR-like cysteine-rich domains and a putative death domain similar to those of TNFR I and Fas. Flow cytometric analysis and immunoprecipitation experiments demonstrated specific binding between the TEF CAR1-related protein and an immunoadhesin composed of the surface (SU) envelope protein of subgroup E (RAV-0) virus fused to the constant region of a rabbit immunoglobulin. These two activities of the TEF CAR1-related protein, specific binding to ALV-E SU and permitting entry only of ALV-E, have unambiguously identified this protein as a cellular receptor specific for subgroup E ALV. PMID- 9326660 TI - Bordetella bronchiseptica dermonecrotizing toxin induces reorganization of actin stress fibers through deamidation of Gln-63 of the GTP-binding protein Rho. AB - Bordetella dermonecrotizing toxin causes assembly of actin stress fibers and focal adhesions in some cultured cells and induces mobility shifts of the small GTP-binding protein Rho on electrophoresis. We attempted to clarify the molecular basis of the toxin action on Rho. Analysis of the amino acid sequence of toxin treated RhoA revealed the deamidation of Gln-63 to Glu. The substitution of Glu for Gln-63 of RhoA by site-directed mutagenesis caused a mobility shift on electrophoresis, which was indistinguishable from that of the toxin-treated RhoA. Neither mutant RhoA-bearing Glu-63 nor toxin-treated RhoA significantly differed from untreated wild type RhoA in guanosine 5'-[gamma-thio]triphosphate binding activity but both showed a 10-fold reduction in GTP hydrolysis activity relative to untreated RhoA. C3H10T1/2 cells transfected with cDNA of the mutant RhoA bearing Glu-63 showed extensive formation of actin stress fibers similar to the toxin-treated cells. These results indicate that the toxin catalyzes deamidation of Gln-63 of Rho and renders it constitutively active, leading to formation of actin stress fibers. PMID- 9326661 TI - Ca2+-independent inhibition of inositol trisphosphate receptors by calmodulin: redistribution of calmodulin as a possible means of regulating Ca2+ mobilization. AB - The interactions between calmodulin, inositol 1,4,5-trisphosphate (InsP3), and pure cerebellar InsP3 receptors were characterized by using a scintillation proximity assay. In the absence of Ca2+, 125I-labeled calmodulin reversibly bound to multiple sites on InsP3 receptors and Ca2+ increased the binding by 190% +/- 10%; the half-maximal effect occurred when the Ca2+ concentration was 184 +/- 14 nM. In the absence of Ca2+, calmodulin caused a reversible, concentration dependent (IC50 = 3.1 +/- 0.2 microM) inhibition of [3H]InsP3 binding by decreasing the affinity of the receptor for InsP3. This effect was similar at all Ca2+ concentrations, indicating that the site through which calmodulin inhibits InsP3 binding has similar affinities for calmodulin and Ca2+-calmodulin. Calmodulin (10 microM) inhibited the Ca2+ release from cerebellar microsomes evoked by submaximal, but not by maximal, concentrations of InsP3. Tonic inhibition of InsP3 receptors by the high concentrations of calmodulin within cerebellar Purkinje cells may account for their relative insensitivity to InsP3 and limit spontaneous activation of InsP3 receptors in the dendritic spines. Inhibition of InsP3 receptors by calmodulin at all cytosolic Ca2+ concentrations, together with the known redistribution of neuronal calmodulin evoked by protein kinases and Ca2+, suggests that calmodulin may also allow both feedback control of InsP3 receptors and integration of inputs from other signaling pathways. PMID- 9326662 TI - Decoding temporally encoded sensory input by cortical oscillations and thalamic phase comparators. AB - The temporally encoded information obtained by vibrissal touch could be decoded "passively," involving only input-driven elements, or "actively," utilizing intrinsically driven oscillators. A previous study suggested that the trigeminal somatosensory system of rats does not obey the bottom-up order of activation predicted by passive decoding. Thus, we have tested whether this system obeys the predictions of active decoding. We have studied cortical single units in the somatosensory cortices of anesthetized rats and guinea pigs and found that about a quarter of them exhibit clear spontaneous oscillations, many of them around whisking frequencies ( approximately 10 Hz). The frequencies of these oscillations could be controlled locally by glutamate. These oscillations could be forced to track the frequency of induced rhythmic whisker movements at a stable, frequency-dependent, phase difference. During these stimulations, the response intensities of multiunits at the thalamic recipient layers of the cortex decreased, and their latencies increased, with increasing input frequency. These observations are consistent with thalamocortical loops implementing phase-locked loops, circuits that are most efficient in decoding temporally encoded information like that obtained by active vibrissal touch. According to this model, and consistent with our results, populations of thalamic "relay" neurons function as phase "comparators" that compare cortical timing expectations with the actual input timing and represent the difference by their population output rate. PMID- 9326663 TI - Identification and characterization of a conserved family of protein serine/threonine phosphatases homologous to Drosophila retinal degeneration C. AB - The Drosophila retinal degeneration C (rdgC) gene encodes an unusual protein serine/threonine phosphatase in that it contains at least two EF-hand motifs at its carboxy terminus. By a combination of large-scale sequencing of human retina cDNA clones and searches of expressed sequence tag and genomic DNA databases, we have identified two sequences in mammals [Protein Phosphatase with EF-hands-1 and 2 (PPEF-1 and PPEF-2)] and one in Caenorhabditis elegans (PPEF) that closely resemble rdgC. In the adult, PPEF-2 is expressed specifically in retinal rod photoreceptors and the pineal. In the retina, several isoforms of PPEF-2 are predicted to arise from differential splicing. The isoform that most closely resembles rdgC is localized to rod inner segments. Together with the recently described localization of PPEF-1 transcripts to primary somatosensory neurons and inner ear cells in the developing mouse, these data suggest that the PPEF family of protein serine/threonine phosphatases plays a specific and conserved role in diverse sensory neurons. PMID- 9326664 TI - A dominant form of inherited retinal degeneration caused by a non-photoreceptor cell-specific mutation. AB - We have isolated a dominant mutation, night blindness a (nba), that causes a slow retinal degeneration in zebrafish. Heterozygous nba fish have normal vision through 2-3 months of age but subsequently become night blind. By 9.5 months of age, visual sensitivity of affected fish may be decreased more than two log units, or 100-fold, as measured behaviorally. Electroretinographic (ERG) thresholds of mutant fish are also raised significantly, and the ERG b-wave shows a delayed implicit time. These defects are due primarily to a late-onset photoreceptor cell degeneration involving initially the rods but eventually the cones as well. Homozygous nba fish display an early-onset neuronal degeneration throughout the retina and elsewhere in the central nervous system. As a result, animals develop with small eyes and die by 4-5 days postfertilization (pf). These latter data indicate that the mutation affecting nba fish is not in a photoreceptor cell-specific gene. PMID- 9326665 TI - A human intermediate conductance calcium-activated potassium channel. AB - An intermediate conductance calcium-activated potassium channel, hIK1, was cloned from human pancreas. The predicted amino acid sequence is related to, but distinct from, the small conductance calcium-activated potassium channel subfamily, which is approximately 50% conserved. hIK1 mRNA was detected in peripheral tissues but not in brain. Expression of hIK1 in Xenopus oocytes gave rise to inwardly rectifying potassium currents, which were activated by submicromolar concentrations of intracellular calcium (K0.5 = 0.3 microM). Although the K0.5 for calcium was similar to that of small conductance calcium activated potassium channels, the slope factor derived from the Hill equation was significantly reduced (1.7 vs. 3. 5). Single-channel current amplitudes reflected the macroscopic inward rectification and revealed a conductance level of 39 pS in the inward direction. hIK1 currents were reversibly blocked by charybdotoxin (Ki = 2.5 nM) and clotrimazole (Ki = 24.8 nM) but were minimally affected by apamin (100 nM), iberiotoxin (50 nM), or ketoconazole (10 microM). These biophysical and pharmacological properties are consistent with native intermediate conductance calcium-activated potassium channels, including the erythrocyte Gardos channel. PMID- 9326666 TI - Activation of a caspase 3-related cysteine protease is required for glutamate mediated apoptosis of cultured cerebellar granule neurons. AB - Neurotoxicity induced by overstimulation of N-methyl-D-aspartate (NMDA) receptors is due, in part, to a sustained rise in intracellular Ca2+; however, little is known about the ensuing intracellular events that ultimately result in cell death. Here we show that overstimulation of NMDA receptors by relatively low concentrations of glutamate induces apoptosis of cultured cerebellar granule neurons (CGNs) and that CGNs do not require new RNA or protein synthesis. Glutamate-induced apoptosis of CGNs is, however, associated with a concentration- and time-dependent activation of the interleukin 1beta-converting enzyme (ICE)/CED-3-related protease, CPP32/Yama/apopain (now designated caspase 3). Further, the time course of caspase 3 activation after glutamate exposure of CGNs parallels the development of apoptosis. Moreover, glutamate-induced apoptosis of CGNs is almost completely blocked by the selective cell permeable tetrapeptide inhibitor of caspase 3, Ac-DEVD-CHO but not by the ICE (caspase 1) inhibitor, Ac YVAD-CHO. Western blots of cytosolic extracts from glutamate-exposed CGNs reveal both cleavage of the caspase 3 substrate, poly(ADP-ribose) polymerase, as well as proteolytic processing of pro-caspase 3 to active subunits. Our data demonstrate that glutamate-induced apoptosis of CGNs is mediated by a posttranslational activation of the ICE/CED-3-related cysteine protease caspase 3. PMID- 9326667 TI - Multipotent neural precursors can differentiate toward replacement of neurons undergoing targeted apoptotic degeneration in adult mouse neocortex. AB - Neurons undergoing targeted photolytic cell death degenerate by apoptosis. Clonal, multipotent neural precursor cells were transplanted into regions of adult mouse neocortex undergoing selective degeneration of layer II/III pyramidal neurons via targeted photolysis. These precursors integrated into the regions of selective neuronal death; 15 +/- 7% differentiated into neurons with many characteristics of the degenerated pyramidal neurons. They extended axons and dendrites and established afferent synaptic contacts. In intact and kainic acid lesioned control adult neocortex, transplanted precursors differentiated exclusively into glia. These results suggest that the microenvironmental alterations produced by this synchronous apoptotic neuronal degeneration in adult neocortex induced multipotent neural precursors to undergo neuronal differentiation which ordinarily occurs only during embryonic corticogenesis. Studying the effects of this defined microenvironmental perturbation on the differentiation of clonal neural precursors may facilitate identification of factors involved in commitment and differentiation during normal development. Because photolytic degeneration simulates some mechanisms underlying apoptotic neurodegenerative diseases, these results also suggest the possibility of neural precursor transplantation as a potential cell replacement or molecular support therapy for some diseases of neocortex, even in the adult. PMID- 9326668 TI - Glyceraldehyde-3-phosphate dehydrogenase: nuclear translocation participates in neuronal and nonneuronal cell death. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein levels increase in particulate fractions in association with cell death in HEK293 cells, S49 cells, primary thymocytes, PC12 cells, and primary cerebral cortical neuronal cultures. Subcellular fractionation and immunocytochemistry reveal that this increase primarily reflects nuclear translocation. Nuclear GAPDH is tightly bound, resisting extraction by DNase or salt treatment. Treating primary thymocytes, PC12 cells, and primary cortical neurons with antisense but not sense oligonucleotides to GAPDH prevents cell death. Because cell-death-associated nuclear translocation of GAPDH and antisense protection occur in multiple neuronal and nonneuronal systems, we propose that GAPDH is a general mediator of cell death and uses nuclear translocation as a signaling mechanism. PMID- 9326669 TI - Long-term potentiation activates the GAP-43 promoter: selective participation of hippocampal mossy cells. AB - Perforant path long-term potentiation (LTP) in intact mouse hippocampal dentate gyrus increased the neuron-specific, growth-associated protein GAP-43 mRNA in hilar cells 3 days after tetanus, but surprisingly not in granule cells, the perforant path target. This increase was positively correlated with level of enhancement and restricted to central hilar cells on the side of stimulation. Blockade of LTP by puffing DL-aminophosphonovalerate (APV), an N-methyl-D aspartate (NMDA) receptor blocker into the molecular layer, eliminated LTP induced GAP-43 mRNA elevation in hilar cells. To determine whether the mRNA elevation was mediated by transcription, LTP was studied in transgenic mice bearing a GAP-43 promoter-lacZ reporter gene. Promoter activity as indexed by Transgene expression (PATE) increased as indicated by blue staining of the lacZ gene product, beta-galactosidase. Potentiation induced a blue band bilaterally in the inner molecular layer of the dentate gyrus along the entire septotemporal axis. Because mossy cells are the only neurons in the central hilar zone that project to the inner molecular layer bilaterally along the entire septotemporal axis and LTP-induced activation of PATE in this zone was confined to the side of stimulation, we concluded that mossy cells were unilaterally activated, increasing synthesis of beta-galactosidase, which was transported bilaterally. Neither granule cells nor pyramidal cells demonstrated increased PATE or increased GAP-43 mRNA levels. These results and recent evidence indicating the necessity of hilar neurons for LTP point to previously unheralded mossy cells as potentially critical for perforant path LTP and the GAP-43 in these cells as important for LTP persistence lasting days. PMID- 9326670 TI - Massive targeting of liposomes, surface-modified with anionized albumins, to hepatic endothelial cells. AB - Human serum albumin (HSA) derivatized with cis-aconitic anhydride was covalently coupled to liposomes with a size of approximately 100 nm [polyaconitylated HSA (Aco-HSA) liposomes]. Within 30 min after injection into a rat, Aco-HSA liposomes were completely cleared from the blood and almost exclusively taken up by the liver, whereas in control liposomes 80% was still present in the blood at that time. Endothelial cells were shown to account for almost two-thirds of the hepatic uptake of the Aco-HSA liposomes, the remainder being recovered mainly in the liver macrophages (Kupffer cells). With fluorescently labeled liposomes it was shown that the Aco-HSA liposomes target a vast majority (>85%) of the cells in the endothelial cell population. Control liposomes were not taken up to a significant extent by the endothelial cells. Uptake of Aco-HSA liposomes by both endothelial and Kupffer cells was inhibited by preinjection with polyinosinic acid, indicating the involvement of scavenger receptors in the uptake process. The uptake of Aco-HSA liposomes by liver endothelial cells was dependent on liposome size; with increasing liposome diameter endothelial cell uptake decreased in favor of Kupffer cell uptake. We have demonstrated that massive in vivo targeting of liposomes to a defined cell population other than macrophages is possible. Aco-HSA liposomes thus may represent an attractive drug carrier system for treatment of various liver or liver endothelium-associated disorders. PMID- 9326671 TI - Peptide analogue studies of the hypothalamic neuropeptide Y receptor mediating pituitary adrenocorticotrophic hormone release. AB - Hypothalamic neuropeptide Y (NPY) is thought to be important in the regulation of feeding and also in the release of Adrenocorticotrophic hormone (ACTH). Intracerebroventricular administration of NPY to male rats significantly increased plasma ACTH 10 min after injection and stimulated 2-h food intake. A series of analogues of NPY that have a greatly reduced affinity for the Y1 [human pancreatic polypeptide (human PP), NPY(3-36)], the Y2 ([Pro34]NPY, human PP), the Y3 (peptide YY), and the Y6 (human PP) receptor, all markedly stimulated ACTH release. Rat PP, which binds with high affinity to the Y4 receptor, was unable to stimulate ACTH release. A novel analogue fragment [Pro34]NPY(13-36) was synthesized as a ligand with low Y1 and Y2 receptor affinity. Interestingly, neither [Pro34]NPY(13-36) nor the selective Y5 receptor agonist [D-Trp32]NPY stimulated food intake, whereas both significantly increased plasma ACTH. Thus the hypothalamic NPY receptor mediating increases in plasma ACTH has a fragment activation profile unlike the Y1-Y4 or Y6 receptors and appears distinct from the NPY receptor controlling food intake. PMID- 9326672 TI - A peptide derived from an extracellular domain selectively inhibits receptor internalization: target sequences on insulin and insulin-like growth factor 1 receptors. AB - Certain peptides derived from the alpha1 domain of the major histocompatibility class I antigen complex (MHC-I) inhibit receptor internalization, increasing the steady-state number of active receptors on the cell surface and thereby enhancing the sensitivity to hormones and other agonists. These peptides self-assemble, and they also bind to MHC-I at the same site from which they are derived, suggesting that they could bind to receptor sites with significant sequence similarity. Receptors affected by MHC-I peptides do, indeed, have such sequence similarity, as illustrated here by insulin receptor (IR) and insulin-like growth factor-1 receptor. A synthetic peptide with sequence identical to a certain extracellular receptor domain binds to that receptor in a ligand-dependent manner and inhibits receptor internalization. Moreover, each such peptide is selective for its cognate receptor. An antibody to the IR peptide not only binds to IR and competes with the peptide but also inhibits insulin-dependent internalization of IR. These observations, and binding studies with deletion mutants of IR, indicate that the sequence QILKELEESSF encoded by exon 10 plays a key role in IR internalization. Our results illustrate a principle for identifying receptor-specific sites of importance for receptor internalization, and for enhancing sensitivity to hormones and other agonists. PMID- 9326673 TI - Regulation of the human ether-a-gogo related gene (HERG) K+ channels by reactive oxygen species. AB - Human ether-a-gogo related gene (HERG) K+ channels are key elements in the control of cell excitability in both the cardiovascular and the central nervous systems. For this reason, the possible modulation by reactive oxygen species (ROS) of HERG and other cloned K+ channels expressed in Xenopus oocytes has been explored in the present study. Exposure of Xenopus oocytes to an extracellular solution containing FeSO4 (25-100 microM) and ascorbic acid (50-200 microM) (Fe/Asc) increased both malondialdehyde content and 2',7'-dichlorofluorescin fluorescence, two indexes of ROS production. Oocyte perfusion with Fe/Asc caused a 50% increase of the outward K+ currents carried by HERG channels, whereas inward currents were not modified. This ROS-induced increase in HERG outward K+ currents was due to a depolarizing shift of the voltage-dependence of channel inactivation, with no change in channel activation. No effect of Fe/Asc was observed on the expressed K+ currents carried by other K+ channels such as bEAG, rDRK1, and mIRK1. Fe/Asc-induced stimulation of HERG outward currents was completely prevented by perfusion of the oocytes with a ROS scavenger mixture (containing 1,000 units/ml catalase, 200 ng/ml superoxide dismutase, and 2 mM mannitol). Furthermore, the scavenger mixture also was able to reduce HERG outward currents in resting conditions by 30%, an effect mimicked by catalase alone. In conclusion, the present results seem to suggest that changes in ROS production can specifically influence K+ currents carried by the HERG channels. PMID- 9326674 TI - Oxytocin releases atrial natriuretic peptide by combining with oxytocin receptors in the heart. AB - Previous studies indicated that the central nervous system induces release of the cardiac hormone atrial natriuretic peptide (ANP) by release of oxytocin from the neurohypophysis. The presence of specific transcripts for the oxytocin receptor was demonstrated in all chambers of the heart by amplification of cDNA by the PCR using specific oligonucleotide primers. Oxytocin receptor mRNA content in the heart is 10 times lower than in the uterus of female rats. Oxytocin receptor transcripts were demonstrated by in situ hybridization in atrial and ventricular sections and confirmed by competitive binding assay using frozen heart sections. Perfusion of female rat hearts for 25 min with Krebs-Henseleit buffer resulted in nearly constant release of ANP. Addition of oxytocin (10(-6) M) significantly stimulated ANP release, and an oxytocin receptor antagonist (10(-7) and 10(-6) M) caused dose-related inhibition of oxytocin-induced ANP release and in the last few minutes of perfusion decreased ANP release below that in control hearts, suggesting that intracardiac oxytocin stimulates ANP release. In contrast, brain natriuretic peptide release was unaltered by oxytocin. During perfusion, heart rate decreased gradually and it was further decreased significantly by oxytocin (10(-6) M). This decrease was totally reversed by the oxytocin antagonist (10(-6) M) indicating that oxytocin released ANP that directly slowed the heart, probably by release of cyclic GMP. The results indicate that oxytocin receptors mediate the action of oxytocin to release ANP, which slows the heart and reduces its force of contraction to produce a rapid reduction in circulating blood volume. PMID- 9326675 TI - A mouse model for the renal salt-wasting syndrome pseudohypoaldosteronism. AB - Aldosterone-dependent epithelial sodium transport in the distal nephron is mediated by the absorption of sodium through the highly selective, amiloride sensitive epithelial sodium channel (ENaC) made of three homologous subunits (alpha, beta, and gamma). In human, autosomal recessive mutations of alpha, beta, or gammaENaC subunits cause pseudohypoaldosteronism type 1 (PHA-1), a renal salt wasting syndrome characterized by severe hypovolemia, high plasma aldosterone, hyponatremia, life-threatening hyperkaliemia, and metabolic acidosis. In the mouse, inactivation of alphaENaC results in failure to clear fetal lung liquid at birth and in early neonatal death, preventing the observation of a PHA-1 renal phenotype. Transgenic expression of alphaENaC driven by a cytomegalovirus promoter in alphaENaC(-/-) knockout mice [alphaENaC(-/-)Tg] rescued the perinatal lethal pulmonary phenotype and partially restored Na+ transport in renal, colonic, and pulmonary epithelia. At days 5-9, however, alphaENaC(-/-)Tg mice showed clinical features of severe PHA-1 with metabolic acidosis, urinary salt wasting, growth retardation, and 50% mortality. Adult alphaENaC(-/-)Tg survivors exhibited a compensated PHA-1 with normal acid/base and electrolyte values but 6 fold elevation of plasma aldosterone compared with wild-type littermate controls. We conclude that partial restoration of ENaC-mediated Na+ absorption in this transgenic mouse results in a mouse model for PHA-1. PMID- 9326676 TI - Phentolamine block of KATP channels is mediated by Kir6.2. AB - The ATP-sensitive K+-channel (KATP channel) plays a key role in insulin secretion from pancreatic beta cells. It is closed both by glucose metabolism and the sulfonylurea drugs that are used in the treatment of noninsulin-dependent diabetes mellitus, thereby initiating a membrane depolarization that activates voltage-dependent Ca2+ entry and insulin release. The beta cell KATP channel is a complex of two proteins: Kir6.2 and SUR1. The former is an ATP-sensitive K+ selective pore, whereas SUR1 is a channel regulator that endows Kir6.2 with sensitivity to sulfonylureas. A number of drugs containing an imidazoline moiety, such as phentolamine, also act as potent stimulators of insulin secretion, but their mechanism of action is unknown. We have used a truncated form of Kir6.2, which expresses independently of SUR1, to show that phentolamine does not inhibit KATP channels by interacting with SUR1. Instead, our results argue that phentolamine may interact directly with Kir6.2 to produce a voltage-independent reduction in channel activity. The single-channel conductance is unaffected. Although the ATP molecule also contains an imidazoline group, the site at which phentolamine blocks is not identical to the ATP-inhibitory site, because phentolamine block of an ATP-insensitive mutant (K185Q) is normal. KATP channels also are found in the heart where they are involved in the response to cardiac ischemia: they also are blocked by phentolamine. Our results suggest that this may be because Kir6.2, which is expressed in the heart, forms the pore of the cardiac KATP channel. PMID- 9326677 TI - Developmental changes in DNA methylation of the two tobacco pollen nuclei during maturation. AB - Changes in DNA methylation during tobacco pollen development have been studied by confocal fluorescence microscopy using a monoclonal anti-5-methylcytosine (anti m5C) antibody and a polyclonal anti-histone H1 (anti-histone) antibody as an internal standard. The specificity of the anti-m5C antibody was demonstrated by a titration series against both single-stranded DNA and double-stranded DNA substrates in either the methylated or unmethylated forms. The antibody was found to show similar kinetics against both double- and single-stranded DNA, and the fluorescence was proportional to the amount of DNA used. No signal was observed with unmethylated substrates. The extent of methylation of the two pollen nuclei remained approximately constant after the mitotic division that gave rise to the vegetative and generative nuclei. However, during the subsequent development of the pollen, the staining of the generative nucleus decreased until it reached a normalized value of (1)/(5) of that of the vegetative nucleus. The use of a confocal microscope makes these data independent of possible focusing artefacts. The anti-histone antibody was used as a control to show that, while the antibody staining directed against 5-methylcytosine changed dramatically during pollen maturation, the histone signal did not. We observed the existence of structural dimorphism amongst tobacco pollen grains, the majority having three pollen apertures and the rest with four. However, the methylation changes observed occurred to the same extent in both subclasses. PMID- 9326678 TI - T-strand integration in maize protoplasts after codelivery of a T-DNA substrate and virulence genes. AB - We describe a plant protoplast transformation method that provides transformants with a simple pattern of integration of a foreign gene. The approach is to deliver into plant protoplasts by direct gene transfer the Agrobacterium virulence genes virD1 and virD2 with or without virE2, together with a target plasmid containing a gene of interest flanked by Agrobacterium T-DNA border repeat sequences of 25 bp. We present evidence of T-DNA formation in maize protoplasts and its integration into the maize genome. The frequency of VirD1 VirD2-mediated integration events was about 20-35% of the total number of transformants. The addition of virE2 doubled the transformation efficiency. The method described here is of sufficient efficiency and simplicity to be useful for the production of transgenic plants with single-copy well-defined transgenic inserts. PMID- 9326679 TI - Arabidopsis thaliana mutants altered in homologous recombination. AB - Homologous recombination contributes both to the generation of allelic diversity and to the preservation of genetic information. In plants, a lack of suitable experimental material has prevented studies of the regulatory and enzymatic aspects of recombination in somatic and meiotic cells. We have isolated nine Arabidopsis thaliana mutants hypersensitive to x-ray irradiation (xrs) and examined their recombination properties. For the three xrs loci described here, single recessive mutations were found to confer simultaneous hypersensitivities to the DNA-damaging chemicals mitomycin C (MMCs) and/or methyl methanesulfonate (MMSs) and alterations in homologous recombination. Mutant xrs9 (Xrays, MMSs) is reduced in both somatic and meiotic recombination and resembles yeast mutants of the rad52 epistatic group. xrs11 (Xrays, MMCs) is deficient in the x-ray-mediated stimulation of homologous recombination in somatic cells in a manner suggesting a specific signaling defect. xrs4 (Xrays, MMSs, MMCs) has a significant deficiency in somatic recombination, but this is accompanied by meiotic hyper-recombination. A corresponding phenotype has not been reported in other systems and thus this indicates a novel, plant-specific regulatory circuit linking mitotic and meiotic recombination. PMID- 9326680 TI - The heme oxygenase gene (pbsA) in the red alga Rhodella violacea is discontinuous and transcriptionally activated during iron limitation. AB - Heme oxygenase (HO) catalyzes the opening of the heme ring with the release of iron in both plants and animals. In cyanobacteria, red algae, and cryptophyceae, HO is a key enzyme in the synthesis of the chromophoric part of the photosynthetic antennae. In an attempt to study the regulation of this key metabolic step, we cloned and sequenced the pbsA gene encoding this enzyme from the red alga Rhodella violacea. The gene is located on the chloroplast genome, split into three distant exons, and is presumably expressed by a trans-splicing mechanism. The deduced polypeptide sequence is homologous to other reported HOs from organisms containing phycobilisomes (Porphyra purpurea and Synechocystis sp. strain PCC 6803) and, to a lesser extent, to vertebrate enzymes. The expression is transcriptionally activated under iron deprivation, a stress condition frequently encountered by algae, suggesting a second role for HO as an iron mobilizing agent in photosynthetic organisms. PMID- 9326681 TI - Feature integration in pattern perception. AB - The human visual system is able to effortlessly integrate local features to form our rich perception of patterns, despite the fact that visual information is discretely sampled by the retina and cortex. By using a novel perturbation technique, we show that the mechanisms by which features are integrated into coherent percepts are scale-invariant and nonlinear (phase and contrast polarity independent). They appear to operate by assigning position labels or "place tags" to each feature. Specifically, in the first series of experiments, we show that the positional tolerance of these place tags in foveal, and peripheral vision is about half the separation of the features, suggesting that the neural mechanisms that bind features into forms are quite robust to topographical jitter. In the second series of experiment, we asked how many stimulus samples are required for pattern identification by human and ideal observers. In human foveal vision, only about half the features are needed for reliable pattern interpolation. In this regard, human vision is quite efficient (ratio of ideal to real approximately 0.75). Peripheral vision, on the other hand is rather inefficient, requiring more features, suggesting that the stimulus may be relatively underrepresented at the stage of feature integration. PMID- 9326682 TI - Ape-like or hominid-like? The positional behavior of Oreopithecus bambolii reconsidered. AB - Comparative morphological and functional analyses of the skeletal remains of Oreopithecus bambolii, a hominoid from the Miocene Mediterranean island of Tuscany-Sardinia (Italy), provides evidence that bipedal activities made up a significant part of the positional behavior of this primate. The mosaic pattern of its postcranial morphology is to some degree convergent with that of Australopithecus and functionally intermediate between apes and early hominids. Some unique traits could have been selected only under insular conditions where the absence of predators and the limitation of trophic resources play a crucial role in mammalian evolution. PMID- 9326683 TI - Prevalence of idiopathic long QT syndrome in children with congenital deafness. AB - Long QT syndrome (LQTS) is characterized by prolongation of the QT interval associated with a high risk for syncope and sudden death. Jervell and Lange Nielsen initially described LQTS in association with congenital sensorineural deafness. We have investigated the prevalence of this syndrome in a school for deaf children, evaluating by ECG 350 congenitally deaf children with an age range of 6-19 years. The corrected QT interval (QTc) was calculated by Bazett's formula. Eight children with a QTc interval >440 ms were further studied by cardiac examination, repeat ECGs (three times), Holter monitoring, echocardiography, and exercise testing. The families were assessed for a history of syncope and deafness and underwent ECG evaluations regarding lengthened QTc interval. Among these eight children only two girls aged 14 and 15 years were diagnosed as having LQTS according to Schwartz's criteria (0.57% of the 350 deaf children; 95% confidence intervals 0, 0.05) in any of these parameters when compared with nonexpanded irradiated tissue. This study demonstrates that radiation produced a significant change in porcine skin, but tissue expansion did not further alter the histological changes associated with irradiation. PMID- 9326711 TI - Minoxidil and wound contraction. AB - Minoxidil has been proposed as a potential topical inhibitor of wound contraction and proliferative scarring. Suggestions for this application are derived from in vitro investigations demonstrating inhibition of various fibroblastic functions. The purpose of this study was to attempt to establish in vivo support of these effects using an established animal model of wound contraction. Standardized cutaneous wounds were created on the dorsum of Sprague-Dawley rats, which were divided equally into six treatment groups. Wounds were treated daily after tracing their unhealed areas. On complete closure of the wounds, analyses of the contraction rates and tensile strength were performed for comparison among groups. Minoxidil did not demonstrate significant inhibition of wound contraction rates relative to either an inert vehicle, an active vehicle, or no treatment. Contrarily, as previously demonstrated in this animal model, silver sulfadiazine did demonstrate significant inhibition of wound contraction rates relative to both vehicles. No significant difference in tensile strength was demonstrated among groups. These observations do not support the proposed use of minoxidil as an "antifibrotic" agent. PMID- 9326712 TI - Sentinel node excision for the diagnosis of metastatic neuroendocrine carcinoma of the skin: a case report. AB - The technology of sentinel node lymphoscintigraphy has made it possible to map and identify the lymph nodes draining the site of a primary cutaneous malignancy. This technique is now being used in the treatment of melanoma, and breast and vulvar carcinoma. With melanoma and breast carcinoma, the histology of the sentinel lymph node (SLN) has been found to be reflective of the histology of the remainder of the nodal basin. The concept of sampling the SLN is to provide an accurate staging for the entire nodal basin, obviating the need for a complete lymphadenectomy if the SLN is negative. It is believed that cutaneous malignancies with a propensity for regional metastasis, such as neuroendocrine carcinoma of the skin, may spread via a similar lymphatic pathway involving an SLN. Using this technique we identified and excised the SLNs in a patient with a neuroendocrine carcinoma of the skin that contained the only focus of metastatic disease. Although this technique is still investigational we believe it holds great promise in being able to detect occult metastatic nodal disease in clinically node-negative patients. PMID- 9326713 TI - Case report: sequential vascular connection of free flaps in the upper extremity. AB - Devastating hand injuries often require multiple microvascular reconstructions. We report a patient in whom two flaps were used for late reconstruction of a devastating hand injury involving devascularization of the right hand, severely comminuted fractures of the hand and forearm, and multiple tendon avulsions. We believe the sequential vascular connection of free flaps offers the best method of reconstruction in this severe case, allowing composite tissue transfer, monitoring of the osseous flap, and optimal positioning of the two free tissue transfers. PMID- 9326714 TI - Management of a large arteriovenous fistula in the face: a case of neurofibromatosis type 1. AB - Neurofibromatosis type 1 (von Recklinghausen's disease or peripheral neurofibromatosis) is of particular interest to the plastic surgeon, as it affects the skin. The majority of affected patients require operative procedures for cosmetic and functional reasons. Rarely, vascular lesions such as stenosis, rupture of an aneurysm, and fistula formation in neurofibromatosis type 1 can create some difficulties during operation without any symptoms before surgery. We report the management of a large occipitojugular fistula. This life-threatening fistula was located on the right side of the face of a 28-year-old patient who had been operated six times for the same neurofibromatous mass without any operative complications. During the operation for partial excision of the neurofibromatous mass, life-threatening, uncontrollable hemorrhage began and the operation was ended without excision. Postoperative angiography revealed an arteriovenous fistula between the occipital artery and jugular vein, and also total occlusion/agenesis in the postclinoid cisternal segment of the internal carotid artery. The fistula was obliterated with coil and histoacryl lipiodol mixture. After this procedure partial excision was performed without abnormal bleeding. PMID- 9326715 TI - Giant cell tumor of the proximal phalanx: a case report. AB - We report a 45-year-old man who presented with a 3-month history of pain and swelling in the proximal phalanx of the right fifth finger. After biopsy and pathological examination the lesion was found to be an aggressive (Enneking's stage 3) giant cell tumor of the proximal phalanx. Due to the high rate of recurrence of stage 3 giant cell tumors in the hand after en bloc resection, it was decided to amputate the right little finger through the metacarpo-phalangeal joint. After a 3-year follow-up of the patient there have been no recurrences or metastases. PMID- 9326716 TI - A historical review of hemostasis, thrombosis, and antithrombotic therapy. PMID- 9326717 TI - Resident education--a casualty of managed care? PMID- 9326719 TI - Re: Selective use of preoperative lower extremity arteriography and free flap reconstruction. PMID- 9326718 TI - Re: Tumescent miniabdominoplasty. PMID- 9326720 TI - Simple securing of subcuticular running suture. PMID- 9326721 TI - Ocular-specific delivery of timolol by sequential bioactivation of its oxime and methoxime analogs. AB - S-(-)- Timolol maleate was oxidized, using the modified Pfitzner-Mofatt method, to the corresponding keto analog, which was then coupled with either hydroxylamine or methoxyamine in the same reaction medium. The products separated, timolone oxime (TO) or timolone methoxime (TMO), were found to be a mixture of both E and Z isomers with the Z isomer in higher concentration. Both isomers could be separated on silica column. No isomerization of any of the isomers could be detected whether in buffers or biological fluids. TMO salts were found to be stable in slightly acidic buffer. The Z isomer of TMO is more stable than the E isomer. Both TO and TMO showed pronounced reduction of the intraocular pressure (IOP) in normotensive rabbits, when instilled into the conjunctival sac. Reduction of IOP caused by either TO or TMO was higher than the reduction produced with the same dose of timolol maleate. Equal doses of any of the TMO isomers or the mixture of isomers gave almost the same percent reduction of IOP. TMO and TO did not show cardiovascular effects when administered intravenously to rabbits or rats. Both are good candidates to be used for topical management of glaucoma without producing systemic side effects. PMID- 9326722 TI - Neuroanatomical aspects of mydriatic action of morphine in rats. AB - Morphine causes mydriasis in rats. In order to investigate whether this effect is due to direct inhibition of preganglionic pupilloconstrictor neurons in the Edinger-Westphal nucleus (EWN), we injected opiate agonists into the EWN in male albino Charles River rats. Bilateral stereotactic microinjections of morphine (10, 20, 30, 40 micrograms/side) inhibited spinal nociceptive reflexes and caused pronounced catalepsy, but had no effect on pupillary size. The powerful opiate agonist, fentanyl, also elicited analgesia and catalepsy, when given in doses of 5 and 10 micrograms/side, but no dose of fentanyl up to 10 micrograms/side induced mydriasis. Naloxone (10 micrograms/side), given into the EWN, effectively antagonized inhibition of the tail-flick response induced by subcutaneously administered morphine (30 micrograms/kg), but had no effect on the cataleptic and mydriatic actions of systemic morphine. These results indicate that, in the rat, morphine-induced mydriasis is not accounted for by a direct action on the EWN. PMID- 9326723 TI - The sensitivity of bovine corneal epithelial lyso-PAF acetyltransferase to cyclooxygenase and lipoxygenase inhibitors is independent of arachidonate metabolites. AB - Lyso-platelet-activating factor (PAF) acetyltransferase is a critical regulatory step in PAF synthesis. PAF accumulates in the cornea in response to injury and is a potent inflammatory mediator that stimulates corneal cyclooxygenase but not lipoxygenase reactions. 12(S)-hydroxyeicosatetraenoic (HETE) acid, a major lipoxygenase product of mammalian corneas, is also generated during injury. In the bovine corneal epithelium, both PAF acetyltransferase and 12-lipoxygenase are microsomal enzymes. A potential interaction between these two lipid mediators was, therefore, examined. PAF acetyltransferase activity was assayed by determining radioactivity in the trichloroacetic acid-precipitated complex of [3H]PAF bound to albumin, formed after incubation of corneal epithelial microsomes with lyso-PAF and [3H]acetyl CoA. Lipoxygenase metabolism by bovine corneal epithelial microsomes was also studied. While 12(S)-HETE did not activate lyso-PAF acetyltransferase, PAF synthesis was decreased when microsomes were treated with lipoxygenase inhibitors. The IC50 values for nordihydroguaiaretic acid (NDGA), and baicalein were 75 and 105 microM, respectively. The IC50 value for CDC, a more potent inhibitor of platelet 12-lipoxygenase, was greater than 200 microM. The results indicate that concentrations suppressing lyso-PAF acetyltransferase activity exceed those required to inhibit lipoxygenases from bovine corneal epithelia. Similarly, concentrations of aspirin and indomethacin, cyclooxygenase inhibitors that decrease PAF formation, were greater than those reported to block prostaglandin generation. A number of other compounds, some structurally similar to the lipoxygenase inhibitors that suppress lyso-PAF acetyltransferase, and others unrelated chemically but known as anti-oxidant and cationic-chelators, also inhibited lyso-PAF acetyltransferase. This suggests that lyso-PAF acetyltransferase activity is inhibited by mechanisms independent of lipoxygenase and cyclooxygenase metabolites. PMID- 9326724 TI - Antagonism of interleukin-1 (IL-1)-induced uveitis with synthetic IL-1 blockers. AB - Because of the discovery of potent interleukin-1 (IL-1) blocking effects by CK 103A (4,6-dihydropyridazino[4,5-c]pyridazin-5 (1H)-ones) on rat uveitis induced by IL-1, numerous derivatives of CK-103A have been synthesized and their efficacies on the same animal model studied. The uveitis was induced by injection of 1 ng IL-1/10 microliters intravitreally. The inflammation reached peak at 12 hr after the injection of IL-1. The prevention/blockade of IL-1-induced uveitis was measured at this peak inflammation time point. It was found that 8 out of 12 CK-analogs studied produced an effective blockade of IL-1-induced uveitis. Most of them were at least equipotent or even more potent than prednisolone in blocking IL-1-induced uveitis. It is concluded that most dihydropyridazinopyridazin derivatives are effective anti-uveitis compounds. Some could be found to be safe and useful for the treatment of this dreadful disease. PMID- 9326725 TI - Potent inhibitory effect of tetrandrine on experimental allergic conjunctivitis in mice. AB - This study investigated the effectiveness of tetrandrine (TDR) on experimental allergic conjunctivitis secondary to ragweed pollen. SWR/J mice were divided as follows: group 1, normal controls; group 2, sensitized but untreated; group 3, sensitized, buffered saline (BS)-treated; and group 4, sensitized, TDR-treated. The last three groups were exposed to ragweed through topical contact on the nasal and conjunctival mucosae followed by challenge with the allergen on the conjunctiva. Groups 3 and 4 received doses of BS and TDR, respectively. The allergic conjunctivitis was evaluated by scoring of the clinical signs and histopathology. mRNA gene expression of interleukin 1 beta (IL-1 beta) and IL-5 in the conjunctiva was analyzed by polymerase chain reaction techniques. All mice exposed to ragweed developed allergic conjunctivitis clinically and histologically. The conjunctivitis was significantly modulated by intraperitoneal injection of a new anti-inflammatory agent, TDR. Histopathologic analysis demonstrated that TDR strikingly reduced the conjunctival eosinophil infiltration and the number of intact and degranulating mast cells. IL-1 beta and Il-5 mRNA gene expression in the conjunctiva of TDR-treated mice was dramatically down regulated compared with untreated and BS-treated controls. The results indicate that TDR may have potential clinical use in the treatment of conjunctivitis. PMID- 9326726 TI - Drug distribution in the vitreous humor of the human eye: the effects of aphakia and changes in retinal permeability and vitreous diffusivity. AB - The majority of eyes that receive drug therapy exhibit some form of pathophysiological degradation. Two common pathophysiological states are retinal inflammation, which results in breakdown of the blood-retinal barrier, and aphakia. The purpose of this study was to examine the effects of aphakia and changes in retinal permeability and vitreous diffusivity on drug distribution in the vitreous humor of the human eye. The study was performed using a finite element model that accurately accounts for the vitreous geometry and boundary conditions. Intravitreal injection is the most common method for treating posterior segment disorders; therefore, this administration method was simulated using the models. Elimination from aphakic and phakic eyes was compared for four extreme injection locations and for two retinal permeabilities. When the retinal permeability was fixed at 5.0 x 10(-5) cm/s, increasing the drug diffusivity through the vitreous from 5.4 to 10(-7) to 2.4 x 10(-5) cm2/s decreased the half life of drug from 64 hours to 2.7 hours. When the drug diffusivity was fixed at 5.6 x 10(-6) cm2/s, increasing the retinal permeability of the drug from 1.0 x 10(-7) to 1.0 x 10(-4) cm/s decreased the half-life of drug from 44 to 7 hours. Therefore, drug diffusivity and retinal permeability are key factors that influence elimination from the vitreous, and must be considered, particularly if the blood retinal barrier has been compromised. Faster drug elimination was observed in aphakic eyes than in phakic eyes, especially for drugs with a low retinal permeability and injected close to the lens capsule. Injection position is also important if the drug is injected in close proximity to a primary elimination barrier. PMID- 9326727 TI - The regurgitation of large vitreous injections. AB - The same quantity of fluorescein labelled dextran was injected into the vitreous humor of 5 rabbits; in one eye, dissolved in 10 microliters solution and in the other, 100 microliters. The level of fluorescence in the two eyes was compared a few days later and found to average only 12% less with the larger volume. It is concluded that regurgitation through the needle hole can be ignored in spite of the very high intraocular pressure created by the 100 microliter injection. PMID- 9326728 TI - Sulfonamides do not reach the retina in therapeutic amounts after topical application to the cornea. AB - Retinal drug concentrations were measured in treated and fellow eyes following topical administration of two potent and permeable carbonic anhydrase inhibitors, MK-927 and trifluoromethanesulfonamide (CF3SO2NH2) in the rabbit. Administration of 2 or 5 drops 2% MK-927 or 2% CF3SO2NH2 gave at 1 hr 1.5-9 microM drug in the retinas of treated eyes and 1.3-3.7 microM drug in the retinas of the fellow eye. After allowance was made for the blood content of the retinas (1.4-13%) and drug contained in whole blood due to systemic absorption, little (< 1.4 microM) or no net accumulation in retina could be documented. In a second series of experiments animals were treated with 5 mg/kg i.v. of either MK-927 or CF3SO2NH2 in order to fully saturate red cell carbonic anhydrase prior to topical administration of drug. For MK-927 there was a net accumulation in retina 3 hrs following topical administration in both treated and fellow eyes (3-4 microM) which was approximately 1/4 the concentration in plasma, suggesting that some drug had permeated the blood-retinal barrier. Similar results were obtained with CF3SO2NH2, giving no difference between treated and fellow eyes; retinal concentrations were approximately 90% of plasma concentrations. These results suggest that there is no direct access to retina by posterior or uveoscleral drainage of topically applied sulfonamides other than systemically absorbed drug. PMID- 9326729 TI - Reversibility of ethambutol optic neuropathy. AB - Ethambutol hydrochloride is one of the routinely used drugs as the first line of antitubercular agents. The delayed onset of ocular toxicity is usually thought to be reversible following rapid withdrawal of the drug. We collected ten consecutive patients with severe visual defects due to ethambutol toxicity, and these patients had received presumably safe ethambutol dosages. Although ethambutol was stopped immediately in all cases, only five patients (50%) experienced visual improvement after a period of 12 months to 3 years follow-up. The other five patients (50%) had permanent visual impairment without recovery. There were no predisposing or risk factors to contribute to poor visual outcome. In the group over 60 years old, only 20% (1/5) experienced visual improvement; in the group less than 60 years old, 80% (4/5) had some visual recovery, the difference between these two age groups being statistically significant. We need more patient collections to answer whether the older patients with ethambutol optic neuropathy have poor prognoses. Ethambutol optic neuropathy, in our follow up study, is not always reversible, especially in the older population. It may cause permanent visual disability. There is no so-called "safe-dosage". We suggest reconsideration regarding the use of ethambutol as one of the first-line antitubercular drugs, especially in older patients. PMID- 9326730 TI - L-deprenyl protects injured retinal precursor cells in vitro. AB - We evaluated the ability of L-deprenyl, a monoamine oxidase B-inhibitor (MAO-B), to preserve the viability of serum-deprived immortalized retinal precursor cells in vitro. We serum-deprived rat neural retinal ganglion cells immortalized by an incompetent retro virus. We instilled L-deprenyl in concentrations ranging from 0.01 to 100 microM. After 72 hours we performed a cell count of the L-deprenyl cultures and the control (no L-deprenyl instilled) with a hemocytometer and flow cytometry. We used transmission electron microscopy, DNA gel electrophoresis, and flow cytometry to determine the mechanism of cell death. This study showed that all five concentrations of L-deprenyl statistically increased the survival rate of immortalized retinal precursor cells at 72 hours in the serum-deprived medium (P = 0.01, ANOVA test). Ten microM and higher appeared to provide the greatest immortalized retinal precursor cell survival (12.7 x 10(-4) cells) compared to the control (5.8 x 10(-4) cells). Flow cytometry also demonstrated a higher percentage of surviving cells at 10 microM (80%) and 100 microM (76%) than with the control (58%) (P = 0.0017, chi2 test). Transmission electron microscopy, DNA electrophoresis, and flow cytometry showed that the mode of cell death was by apoptosis. This study suggests that L-deprenyl may be worthy of further investigation as a neuroprotective agent to treat chronic open-angle glaucoma. PMID- 9326731 TI - Sex-linked differences in equilibrium reactions among adolescents performing complex sensorimotor tasks. AB - Equilibrium reactions were compared between male and female adolescents (prepuberal and puberal), classified into two groups: those who had previously learned complex motor tasks (dance or acrobatics) and those with no particular training. Subjects stood (eyes open or eyes closed) on a free seesaw platform, the displacements of which were calculated from accelerometer measures. They were instructed to maintain a vertical position with their frontal plane either parallel (to measure antero-posterior oscillations) or perpendicular to the axis of the platform (to measure lateral oscillations). Girls had a better stability than boys as shown by the smaller displacement of their center of gravity. Untrained subjects, irrespective of sex, were the least stable. Subjects trained in acrobatics were more stable than dancers. Differences related to sex can be attenuated by physical training involving equilibrium exercises which suggests that moderate sustained training could reduce the incidence of falls in aged persons and in professionally exposed workers. PMID- 9326733 TI - A proposed mechanism for memory and learning based upon very high frequency signals in the serotonergic neuronal system. AB - Evidence that the serotonergic neuronal system is associated with memory and learning is discussed. It is proposed that the serotonergic neuronal system has the capacity to generate very high oscillatory frequencies which propagate signals along the microtubule cytoskeletal structure of this neuronal system. A mechanism whereby the very high frequencies couple in a co-operative and synchronous fashion with sensory and event related signals in frequency specific channels is described. The specific and discrete frequency channels involve the receptor areas of serotonergic activity in a model of information storage and retrieval. PMID- 9326732 TI - Reversal of progesterone-induced sequential inhibition by progesterone metabolites. AB - Previous reports have shown that intrabrain administration of progesterone (P) ring A-reduced metabolites into the medial preoptic area (MPOA) and ventromedial hypothalamus (VMH) induces facilitation of female sexual behavior in ovariectomized (ovx) rats pretreated with estrogen. Present studies were designed to explore the possibility that ring-A reduced progesterone metabolites might play a role in controlling the duration of estrous behavior. To this aim ovariectomized (ovx) Sprague Dawley rats implanted with guide cannulae directed towards the VMH or the MPOA were submitted to a systemic hormonal treatment to provoke P-induced sequential inhibition (estradiol benzoate (EB) at time O + P at 44 h + P at 68 h). The second dose of P was administered simultaneously with the i.c. implantation of one of the following P metabolites: 3 beta-hydroxy-5 beta pregnan-20-one (5 beta,3 alpha P), 3 alpha-hydroxy-5 beta-pregnan-20-one (5 beta,3 alpha P) or 3 beta-hydroxy-5 beta- pregnan-20-one (5 alpha,3 beta P) into the MPOA or VMH. Lordosis behavior was evaluated by the lordosis quotient (LQ = number of lordosis/10 male mount x 100) and by the percentage of responding subjects. Results show that 5 beta,3 beta P implanted into the VMH or MPOA counteracted the sequential inhibitory effect induced by systemic administration of P.5 alpha,3 beta P was also able to counteract sequential inhibition, but with less potency and only in the VMFI. Results showed that P-induced sequential inhibition can be counteracted by intrabrain administration of ring-A reduced progestins in both the VMH and MPOA. Data are discussed in terms of a putative physiological role of naturally occurring P metabolites in P-mediated female sexual behavior expression. PMID- 9326734 TI - The state of consciousness and very high frequency signalling in the serotonergic neuronal system. AB - The attributes of consciousness are briefly discussed and matched to the known characteristics of the serotonergic neuronal system. In particular sleep, the action of agonist drugs, the regulatory role and monitoring components, together with the receptor areas for serotonergic activity, are identified as significant correlates of consciousness. Frohlich-like phenomena are attributed to the topology of the serotonergic cell which is posited to produce excitations in the gigaHz or very high frequency range. Coding by discrete channel frequency and propagation throughout the vast microtubule cytoskeletal structure of serotonergic activity is proposed as the biophysical basis of consciousness in higher mammals, particularly humans. PMID- 9326735 TI - Population-specific behavioral electrosensitivity of the European blind cave salamander, Proteus anguinus. AB - In nine salamanders from different Slovenian populations of the urodele Proteus anguinus, including three specimens of its 'black' variety, P anguinus parkelj, thresholds of an overt avoidance response to electrical field stimuli were estimated as a function of frequency (continuous sine-waves in water). Thresholds down to 0.3V/cm (ca 100 nA/cm2) and up to 2 mV/cm (670 nA/cm2), at 'best frequencies' of around 30 Hz were found. Sensitivity covered a total frequency range of below 1 Hz, excluding DC, up to 1-2 kHz with up to 40 dB higher thresholds. Thresholds and tuning curves are compared with those of a Proteus population raised in captivity for more than 35 years. The biological significance and the apparently still ongoing evolution of the electrical sense in urodeles, ie in the genus Proteus, are interpreted in terms of comparative sensory physiology and ethological ecology as a result of more recent evolutionary diversification during and since glaciation in the Pleistocene. PMID- 9326736 TI - Reserpine effects on neurotransmitters in chick heart during growth. AB - Effects of tranquilizing agents on neurotransmitters in the heart have not been widely studied. Thus, the effect of intraperitoneal injection of reserpine, (2.5 mg/kg bw) on the concentrations of excitatory (glutamic acid, glutamine, aspartic acid, asparagine), inhibitory (GABA, glycine, alanine, taurine), neurotransmitters as well as the enzymes (GOT and GPT) and total protein were measured in both heart and serum chicks at different ages (1, 7, 30, 90 and 180 days). Reserpine induced a decrease in the excitatory amino acids and an increase in GABA in both heart and serum in most ages. Glycine and alanine increased in the heart and decreased in serum. Taurine increased in the heart of young ages (1 and 7 days) and decreased in older ones (90 and 180 days), however, it decreased in serum of most ages. Both GOT and GPT increased in heart but, in serum, GOT increased and GPT decreased in most ages. Total protein increased in the heart of young chicks and decreased in the 90- and 180-day-old chicks. In conclusion, reserpine induced a parallel decrease in the ratio glutamate, glutamine, aspartate/GABA in both myocardial tissue and serum of the different age groups. Changes observed in neurotransmitters of the heart suggest that these amino acids may play a similar role in the myocardial tissue, as is described in the central nervous system. PMID- 9326737 TI - Effect of different illumination conditions and ionic environment on the guanylate cyclase activity in retina, optic nerve and optic chiasm of the rat. AB - Guanylate cyclase is sensitive to changes of light and dark periods in incubated extracts obtained from soluble fractions of the retina, optic nerve and optic chiasm. The changes in guanylate cyclase activity found, about 100-fold between dark and light periods in those tissues, indicate a key role for this enzyme. The results showed that light inhibits strongly the retinal guanylate cyclase activity, while it increases the activity of this enzyme in the optic nerve. A generalized photo-inhibited response of guanylate cyclase was observed in all studied tissues in animals adapted to the dark. This suggests that light could act as a double stimulus gating the central circuit which promotes the hydrolysis of cGMP via cGMP phosphodiesterase-rhodopsin-transducin cascade, and by direct inhibition of the retinal guanylate cyclase activity. Finally, different responses have been observed in the guanylate cyclase activity in relation with the ion exposure depending on the studied tissue. In summary, all indicate an important role for the soluble guanylate cyclase activity in retina, and other tissues involved in the visual process such as optic nerve and optic chiasm, which have not been examined until now. PMID- 9326738 TI - Effect of rilmenidine injection into the paraventricular nucleus of the hypothalamus on the water intake induced by application of angiotensin II to the subfornical organ. PMID- 9326739 TI - Age-dependent effect of pituitary transplants on immune responses in rat submaxillary lymph nodes: modulatory effect of the autonomic nervous system. AB - An anterior pituitary was grafted into the breast muscles of male rats on day 5, or under the kidney capsule on day 30 or 60 of life. At the 70th day of life (rats operated at the 5th or 30th day) or at the 100th day of life (rats operated at the 60th day), rats were injected subcutaneously with Freund's complete adjuvant, being killed 2 days later. Rats pituitary-grafted at the 30th day showed a greater hyperprolactinemia than rats grafted at the 5th or 60th day. Submaxillary lymph node natural killer (NK) activity decreased in neonatally pituitary-grafted rats and increased in rats grafted at the 30th or 60th day of life, while lymph node cellularity decreased in rats grafted at the 30th or 60th day. In the case of lipopolysaccharide (LPS)-and concanavalin A (Con A)-induced cell proliferation in lymph nodes, the only significant factor detected was age of transplantation, the effect being less evident in older animals. To examine whether the autonomic denervation of submaxillary lymph nodes affected immune responses in pituitary-grafted rats, animals were subjected to a unilateral superior cervical ganglionectomy (Gx) and/or a parasympathetic decentralization (Dc; by chorda tympani section), 10 days before immunization with Freund's adjuvant. A unilateral Gx blunted the stimulation of lymph node NK activity in rats receiving a pituitary transplant at the 30th or 60th day, but not at the 5th day. Only in sympathetically denervated lymph nodes a significant effect of pituitary transplants on LPS mitogenic effect was found, with significantly less effect of LPS in rats pituitary-grafted at the 5th or 30th day of life. Regarding Con A, a unilateral Gx or unilateral Gx plus Dc uncovered a significant depressive effect of pituitary transplants with a significantly smaller Con A mitogenic effect at each studied age in ipsilaterally Gx lymph nodes, and at 30 and 60 days of age in ipsilaterally Gx plus Dc lymph nodes. Pituitary grafts augmented Con A- induced proliferation in submaxillary lymph nodes at the 5th day of life while they decreased it at the 60th day of life. Pituitary transplants augmented cellularity at the sympathetically denervated lymph nodes and decreased it at the contralateral sham-operated side. Pituitary transplants also diminished lymph node cellularity at the Dc side, the effect being significant in rats transplanted at the 30th day of life. The results are compatible with age dependent, inhibitory as well as promoting activities of hyperprolactinemia on immune responses in submaxillary lymph nodes, depending in part on intact autonomic nerves. PMID- 9326740 TI - Intracerebroventricular injection of interleukin-1 suppresses peripheral lymphocyte function in the primate. AB - The cytokine interleukin-1 (IL-1) can act within the brain to induce peripheral endocrine and immune effects. In the rodent intracerebroventricular (i.c.v.) injection of IL-1 activates the hypothalamic-pituitary-adrenal axis and suppresses peripheral immune function by a CRH-dependent mechanism. It is unknown if IL-1 can similarly act within the brain to cause peripheral immunosuppression in the primate and to what extent this could be attributed to the IL-1-induced increase in ACTH and cortisol levels. In this study we have characterized the pituitary-adrenal and peripheral lymphocyte responses to IL-1 alpha (4.2 micrograms) infused over 30 min into the lateral ventricle of ovariectomized monkeys (n = 5) as compared with responses to an intravenous (i.v.) ACTH infusion (1 microgram/h for 7 h; n = 4). Four serial blood samples were obtained for ACTH and cortisol determination and for lymphocyte isolation during a 1-hour baseline and for 7 h after IL-1 or ACTH. Lymphocyte proliferation was measured by 3H thymidine uptake in response to stimulation with phytohemagglutinin. In all 5 animals, IL-1 alpha caused rapid and profound suppression of lymphocyte mitogen responsiveness for 7 h. Baseline lymphocyte proliferation was 51,800 +/- 9,780 cpm and suppressed to a nadir of 4.5% with a mean of 23% baseline over 7 h (p < 0.001). Mean ACTH and cortisol levels increased from 33 +/- (SEM) 4.6 pg/ml and 43 +/- 4.0 micrograms/dl, respectively, during the control period to 90 +/- 14 pg/ml and 56 +/- 2.6 micrograms/dl, respectively, after IL-1 (p < 0.01). Before i.v. ACTH, baseline lymphocyte proliferation was 49,400 +/- 2,820 cpm, and suppressed to a mean of 64% of baseline during ACTH infusion (p < 0.05). Mean ACTH and cortisol levels increased from 48 +/- 5.0 pg/ml and 43 +/- 2.0 micrograms/dl, respectively, to 170 +/- 34 pg/ml and 66 +/- 2.3 micrograms/dl, respectively, during the ACTH infusion (p < 0.01). Lymphocyte suppression after i.c.v. IL-1 was much more profound than after i.v. ACTH (p < 0.01); the area under the IL-1 response curve was 37% of the area under the ACTH response curve. These studies demonstrate for the first time in the primate that centrally injected IL-1 has a profound suppressive effect on lymphocyte function. They also show for the first time in any species that there appears to be a significant immunosuppressive message produced by i.c.v. IL-1 that is not accounted for by the associated increases in ACTH and cortisol. PMID- 9326741 TI - Growth hormone induces interferon gamma production and may play a role in the presentation of alloantigens in vitro. AB - Several reports support the view that growth hormone (GH) promotes proliferation and cytotoxicity by T cells in a mixed leukocyte culture (MLC). The present study was undertaken to begin to determine the mechanism of action of GH on the MLC in vitro. First, we determined that peripheral blood mononuclear cells (PBMC) cultured with mitomycin-treated allogeneic PBMC in an MLC in the presence of exogenously added rhGH develop an augmented proliferative (25-100%) and cytotoxic response (50-600%). We next examined the possibility that GH may promote alloresponses by inducing gamma-interferon (IGN gamma) production. In these experiments, in situ hybridization was used to determine the frequency of cells expressing mRNA for IFN gamma. It was observed that GH increased significantly the frequency of cells expressing mRNA for IFN gamma (100-800%). To determine the site of action of rhGH, we evaluated the response of purified T cells to alloantigens in the presence of rhGH. The addition of rhGH to an MLC had no demonstrable effect when purified T cells were used as the responding population. However, when T cells were reconstituted with autologous mitomycin-treated PBMC and used as the responding population, rhGH augmented proliferation and cytotoxicity. Taken together, these data show that rhGH augments proliferation, cytotoxicity and IFN gamma production during an MLC, and at lease part of the action of rhGH appears to be on the autologous antigen-presenting cell. PMID- 9326742 TI - Early organ-specific hemorrhage-induced increases in tissue cytokine content: associated neurohormonal and opioid alterations. AB - Hemorrhage is associated with an impairment in the immune response and with increased concentrations of circulating inflammatory cytokines. The present study determined the time course and localization of alterations in circulating and tissue pro-inflammatory cytokines (TNF-alpha, IL-1-alpha and -beta) in response to fixed-pressure (40 mm Hg) hemorrhage as well as the associated hanges in circulating neurohormonal and opioid mediators. Conscious unrestrained non heparinized male Sprague-Dawley rats (n = 24) underwent hemorrhage followed by standard resuscitation with lactated Ringer's solution. Animals were sacrificed at three time points; immediately after the hemorrhage period, at completion of resuscitation and 1.5 h after the resuscitation period. Hemorrhage resulted in marked elevations in circulating levels of TNF-alpha, which averaged 860 +/- 201 pg/ml. The levels were similarly elevated following fluid resuscitation (877 +/- 196 pg/ml) and had decreased towards baseline 1.5 h after completion of resuscitation (281 +/- 134 pg/ml). TNF-alpha was not detectable in plasma of time matched controls. Hemorrhage elevated TNF-alpha content in spleen (25%), lung (55%) and heart (20%), and tissue content remained elevated despite resuscitation. No significant changes in tissue content of TNF-alpha were detected in the liver, kidney or brain. Circulating levels of IL1-alpha and -beta were not detectable in either the time-matched controls or hemorrhaged animals. However, statistically significant elevations in tissue content of IL-1 alpha were observed in heart, spleen, lung, gut and whole brain (15-30%). Tissue content of IL-1 beta did not change in response to hemorrhage and/or fluid resuscitation. Activation of sympathetic outflow, as evidenced by a 3- to 4-fold elevation in circulating epinephrine and norepinephrine levels, was observed immediately after hemorrhage, and was associated with a 5-fold rise in circulating beta-endorphin. These results demonstrate an early increase in tissue cytokine content following hemorrhagic shock, which is associated with elevations in circulating catecholamines and endogenous opioids, consistent with their potential modulatory role in this response. PMID- 9326743 TI - A potential mechanism of local anti-inflammatory action of alpha-melanocyte stimulating hormone within the brain: modulation of tumor necrosis factor-alpha production by human astrocytic cells. AB - The proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) occurs in CNS tissue in neurological disorders, infection, and injury. Its excessive production is believed to contribute to local pathology, in which case modulation of TNF-alpha production should promote survival of neural tissue. The neuropeptide alpha-melanocyte stimulating hormone [alpha-MSH (1-13)] inhibits TNF alpha production in vivo and in vitro, and in this research we tested the capacity of the peptide, and of an anti-inflammatory COOH-terminal tripeptide fragment of it, to inhibit TNF-alpha production induced by bacterial endotoxin in cells of a human glioma line (A-172, anaplastic astrocytoma cells). Both peptides were effective, although the alpha-MSH (1-13) sequence was more potent. Preincubation of the cells with alpha-MSH (1-13) markedly increased its effectiveness. The anticytokine effect of alpha-MSH in glioma cells may be mediated by human melanocortin-1 receptors; mRNA for this receptor subtype was isolated from resting A-172 cells. These results, combined with prior evidence of effectiveness of alpha-MSH molecules in modulating inflammatory processes and of their low toxicity, suggest that the molecules may be useful in the treatment of CNS disorders that have an inflammatory component. PMID- 9326744 TI - Substance P as an immune modulator of anxiety. AB - Peripheral blood concentrations of the proinflammatory peptide substance P (SP) have been shown to increase in response to psychological anxiety in human subjects. In this study, we examined changes in SP levels in peripheral blood in response to the anxiety of a diagnostic medical procedure. The levels of SP were found to he higher in subjects with high initial anxiety as compared to subjects with low initial anxiety as measured on the Multiple Affect Adjective Checklist. Changes in the percentages of CD-8-expressing T lymphocytes were found to correlate with alterations in measures of anxiety as well as SP. These changes persisted for 3 days following the diagnostic procedure. The results of the study seem to indicate that SP may serve as a mediator in stress-induced immune reactions. PMID- 9326745 TI - A selective impairment of the IL-2 system in lymphocytes of patients with glioblastomas: increased level of soluble IL-2R and reduced protein tyrosine phosphorylation. AB - The occurrence of brain tumors is associated with broad suppression of the immune system function; however, the mechanisms involved in this impairment are not fully characterized. In this study, we have examined mechanisms involved in diminished T lymphocyte reactivity in patients with glioblastomas as compared to patients with other types of brain tumors. We found that the proliferative response of T lymphocytes stimulated with phytohemagglutinin or anti-CD3 was significantly reduced in these patients as compared to patients with meningiomas, oligodendrogliomas and healthy individuals. Stimulated T cells appear to express lower levels of the alpha-subunit (p55) of the IL-2 receptor (IL-2R), and increased levels of soluble IL-2R in cell supernatants, whereas no significant differences were observed in the level of the beta (p75)- or gamma-subunits. In addition, we found that competent T cells of glioblastoma patients exhibit lower levels of tyrosine phosphorylation in response to IL-2 as compared with cells of healthy donors. The decrease in the levels of IL-2 and its receptor was selective since no significant changes were observed in the secretion of other Th1- and Th2 derived cytokines (IFN-gamma and IL-4) and the expression of their respective receptors. These results indicate that the diminished response of T cells obtained from patients with glioblastomas may be due to a selective defect in the production of IL-2 and in the expression of functional IL-2R due to a decreased expression of the membranal IL-2R alpha and to lower levels of tyrosine phosphorylation in response to IL-2. PMID- 9326746 TI - Neurochemical sensitization in the pathophysiology of schizophrenia: deficits and dysfunction in neuronal regulation and plasticity. AB - Existing pathophysiological models of schizophrenia are limited in their ability to account for all the clinical dimensions of the disorder. The purpose of this article is to describe a comprehensive hypothesis of the pathophysiology of schizophrenia and specifically how a deficit in neural regulation of developmental origin can lead to a pathologic form of neuroplasticity, i.e., neurochemical sensitization, which causes the onset and psychotic symptoms of the illness. We propose that the symptoms of schizophrenia may be caused by deficits in neural regulation resulting in a pathologic condition of neurochemical sensitization analogous to the preclinical model of pharmacologically-induced behavioral sensitization. This condition, if sustained, can lead to potential neurotoxic effects which produce structural neuronal alterations and persistent morbidity. Several lines of indirect and direct clinical evidence are consistent with this hypothesis. These include the ability of stimulant and psychotomimetic drugs to induce psychosis in normal subjects, the development of apparent sensitization to psychosis-inducing effects of stimulants in chronic stimulant abusers and the increased susceptibility of patients with schizophrenia to the psychotogenic effects of Dopamine (DA) agonists. This hypothesis integrates and extends the work of other investigators and is consistent with specific aspects of the longitudinal course of schizophrenia. The association of longer duration and more episodes of psychosis, with poor treatment response and outcome, are also consistent with this model. Form this hypothesis, specific predictions about the illness course, treatment interventions, and pathophysiologic features of schizophrenia can be derived and tested through clinical investigation. PMID- 9326747 TI - Effect of antipsychotics on regional cerebral blood flow measured with positron emission tomography. AB - Positron Emission Tomography (PET) imaging of regional cerebral blood flow (rCBF) provides an in vivo method for studying brain function. We used [15O]H20 PET to assess the effect of antipsychotic medications on rCBF in 17 subjects with schizophrenia. Each subject was scanned while receiving antipsychotic medication, and after having been withdrawn from antipsychotic medication for a 3-week period. The two scans were subtracted from one another, using a within subjects design, and the areas of difference were identified using the Montreal method. Subjects treated with antipsychotic medication had significantly higher rCBF in the left basal ganglia and left fusiform gyrus compared with the "off-medication" condition. Significantly higher relative rCBF in the anterior cingulate, left dorsolateral and inferior frontal cortex, and left and right cerebellum was observed when off antipsychotic medication. Upregulation of dopamine D2 receptors may lead to a regional increase of blood flow and metabolism in the basal ganglia, which may explain recently reported anatomical enlargement in these regions. PMID- 9326748 TI - The effect of fluoxetine on sleep EEG in childhood depression: a preliminary report. AB - Fluoxetine is associated with substantial objective and subjective sleep disturbance in adults with major depressive disorders. In this preliminary report, the effects of fluoxetine on sleep electroencephologram (EEG) are described in 6 children and adolescents with nonpsychotic major depression. Fluoxetine increased light Stage 1 sleep, the number of arousals and rapid eye movement (REM) density. REM latency was largely unaffected. Oculomotor abnormalities were also evident on treatment, accompanied by an increase in myoclonic activity. Subjective sleep was also disturbed on treatment. These results are in keeping with those observed in depressed adults treated with fluoxetine. PMID- 9326749 TI - Effects of antiglucocorticoid treatment on 5-HT1A function in depressed patients and healthy subjects. AB - Clinical studies suggest that 5-HT1A receptor function may be blunted in depression, while 5-HT1A agonists may possess antidepressant activity. Preclinical findings implicate changes in 5-HT1A receptor sensitivity in the mechanism of antidepressant action. The hyperactivity of the hypothalamic pituitary-adrenal (HPA) axis in depression could be related to those observations, since 5-HT1A receptors are inhibited by glucocorticoids. To evaluate the interaction of the HPA and 5-HT1A systems, we pretreated 15 unipolar depressed patients and 12 healthy control subjects with the antiglucocorticoid ketoconazole (KTCZ) prior to administration of a test dose of the 5-HT1A agonist ipsapirone (IPS). Neuroendocrine (ACTH, cortisol, growth hormone), physiological (hypothermia), and behavioral responses to IPS were assessed. As expected, KTCZ inhibited cortisol biosynthesis, but non-HPA responses to IPS were not enhanced. This study failed to show that glucocorticoid modulation of 5-HT1A receptor function is altered in depression. PMID- 9326750 TI - Sumatriptan-induced growth hormone release in patients with major depression, mania, and normal controls. AB - The purpose of this study was to assess serotonergic function in patients with major depression or mania using sumatriptan, a novel 5-HT1D receptor agonist, as a pharmacological probe in a neuroendocrine challenge paradigm. We studied 18 drug free patients (10 with acute unipolar major depression and 8 with acute mania) who met DSM-IV criteria, and healthy controls. Subjects presented for testing after an overnight fast. After obtaining a blood sample for baseline growth hormone (GH) levels, sumatriptan (6 mg) was given subcutaneously, and further blood samples were collected at half hour intervals for 2 hours. The results showed that GH responses to sumatriptan were blunted in depressed patients but not in manics, compared to healthy controls. There were no differences in basal GH levels between the 3 groups. The results of this study provide further support for the role of serotonergic system in pathophysiology of major depression, but not in mania. PMID- 9326752 TI - Analysis of the 5'-flanking promoter region of the 5-HT2A receptor gene in schizophrenia. AB - We studied the 5'-flanking promoter region comprising positions -1441 to +128 of the 5-HT2A receptor gene in 95 schizophrenics and 100 unrelated normal control subjects. The genes encoding the 5-HT2A receptor exons and the adjoining introl regions had already been studied in these subjects, but no disease specific polymorphism was found (Ishigaki et al., 1996). The DNA fragments were amplified by means of the polymerase chain reaction (PCR), and then analyzed by the single stranded conformation polymorphism (SSCP) and sequencing methods. One patient had a substitution, from A to G, at position -668, and a 5 nucleotide deletion of TACTT at positions -646 to -642, however, the patient also had a normal sequence on the other allele. SSCP analysis showed that the other schizophrenics and the control subjects did not have any polymorphism in the 5'-flanking promoter region of the 5-HT2A receptor gene. PMID- 9326751 TI - Depressive response to physostigmine challenge in borderline personality disorder patients. AB - The purpose of this study was to examine the relationship between mood and hormonal responses to cholinergic challenge with physostigmine in order to assess cholinergic system responsiveness in borderline personality disorder (BPD) patients, other non-BPD personality disorder patients, and normal controls. Thirty-four personality disorder patients, 10 of whom met criteria for BPD and 24 of whom met criteria for other, non-borderline, personality disorders, and 11 normal controls participated in a double blind, placebo controlled physostigmine challenge paradigm. The Profile of Mood States depression subscale (POMS-D) self report measure was obtained at baseline and following the physostigmine or placebo infusions. A repeated measures ANOVA of POMS-D scores in placebo and drug conditions indicated a significantly greater depressive response in the total cohort of personality disorder patients than in the normal comparison group (p < 0.05). However, the depressive response to physostigmine was significantly greater in BPD patients, but not other personality disorder patients, compared to normal controls (p < 0.05). There was a correlation between the peak placebo corrected depressive response to physostigmine and a group of BPD traits related to affective instability but not a group of BPD traits related to impulsivity. There was no correlation in any group between mood response to physostigmine and changes in plasma cortisol, prolactin, or growth hormone, or to nausea or other side effects following physostigmine infusion. These data suggest that there is an association between BPD and acute depressive responses to physostigmine challenge, and that the cholinergic system may be involved in the regulation of affect in Axis II disorders. PMID- 9326753 TI - Three CAG trinucleotide repeats on chromosome 6 (D6S1014, D6S1015, and D6S1058) are not expanded in 30 families with schizophrenia. AB - Since 1991, more than five neuro-genetic disorders have been recognized to be caused by trinucleotide repeat expansions, and the list of such disease should grow. The diseases are characterized clinically by the phenomenon of anticipation, i.e., worsening of the disease phenotype in successive generations with increasing trinucleotide repeat expansion. The presence of anticipation in familial schizophrenia has been suggested. Several studies have provided supportive evidence that the suceptability locus for schizophrenia is on chromosome 6. Therefore, we analyzed three CAG trinucleotide repeat clones D6S1014, D6S1015, and D6S1058 on chromosome 6, which are polymolphic in 30 families with schizophrenia. No unusually, long alleles that would suggest abnormal expansion of more than 35 trinucleotide repeats were observed for these genes. Also, no statistically significant differences were found between the offspring and parental generations of affected subjects or between the affected and unaffected subjects in families with schizophrenia. PMID- 9326754 TI - Plasma levels of arginine vasopressin elevated in patients with major depression. AB - Mentally healthy subjects show increased plasma concentrations of the neuropeptides, arginine vasopressin (AVP) and oxytocin (OT), under conditions of stress, but data are lacking about plasma concentrations of AVP and OT in patients with major depression. We thus assessed plasma concentrations of AVP and OT in patients with major depression (n = 52) and healthy controls (n = 37). Mean plasma AVP concentrations were higher in the group of depressed patients than in controls. A subgroup of 16 patients showed very high levels of plasma AVP, but no other feature differentiating this subgroup from the other patients was found. In patients showed higher plasma AVP levels than out-patients, and melancholic patients had higher plasma AVP levels than did nonmelancholic patients. Plasma AVP levels were slightly related to psychomotor retardation and significantly inversely to neuroticism. Patients' plasma OT concentrations had a wider range than in controls. AVP and AVP-mediated functions may be a factor in the clinical picture of depression, possibly by influencing the activity of the hypothalamic pituitary-adrenal axis. PMID- 9326755 TI - [Proceeding of the 12th annual orthopedic research meeting of the Japanese Orthopedic Association. October 16-17, 1997. Niigata. Japan. Abstracts]. PMID- 9326757 TI - Ritualistic female genital mutilation: current status and future outlook. AB - Ritualistic sexual mutilation of females dates back to the fifth century B.C. This traditional practice is a social as well as a health issue that affects the physical and mental well being of the women who undergo it. Although practiced mostly in African countries north of the equator and the Middle-East, concern has recently been expressed that female genital mutilation is also being practiced in the U.S., Europe, and other western countries by immigrants from these countries. This review describes the various types of female genital mutilation and presents the historical and cultural background of the tradition, outlines the medical, psychological and sexual problems, and discusses the current status and future outlook for this tradition, emphasizing social, medical, and legislative aspects. PMID- 9326756 TI - Divine and rational: the reproductive health of women in ancient Egypt. PMID- 9326758 TI - Evidence for magnesium sulfate as a tocolytic agent. AB - The objective of our study is to quantitatively examine the available evidence regarding the efficacy and side effects of magnesium sulfate for acute tocolysis (from randomized trials) compared with placebo and beta-agonist agents. Randomized trials comparing magnesium sulfate with placebo or beta-agonists for tocolysis were identified with a MEDLINE-based search and was supplemented by a search of obstetrical textbooks and bibliographies. Trials underwent quality evaluation and data abstraction by two independent, blinded investigators. Outcomes evaluated included delivery delay of various durations as well as the frequency of major and minor side effects. Summary odds ratios and 95 percent confidence intervals for dichotomous outcomes were calculated using a random effects model. Interstudy heterogeneity for these outcomes was assessed with a Q statistic. We identified 12 randomized controlled trials of magnesium sulfate for acute tocolysis. Four studies were excluded because of either lack of comparison of magnesium sulfate to either placebo or beta-agonists or lack of reporting clinical outcomes of interest. The eight remaining randomized trials comparing magnesium sulfate with placebo or beta-agonists were included in this analysis. There was no significant difference between MgSO4 and placebo for any of the measured outcomes for delay in delivery. Comparing magnesium sulfate to ritodrine or beta-agonists did not demonstrate any differences between the agents in achieving clinically significant tocolysis. There was a significant difference between MgSO4 and beta-agonists in the frequency of medication discontinuation because of side effects, but not in the frequency of major adverse drug events. There are few data comparing magnesium sulfate with a placebo for acute tocolysis. Magnesium sulfate seems to be comparable to ritodrine and beta agonists, although the available data are not sufficient for a rational choice between these agents. PMID- 9326759 TI - Determinations of serum tumor necrosis factor alpha in corneal allografts. AB - Corneal allograft rejection culminates in a series of interactions between different classes of antigen presenting cells, cytokines and leukocytes. Tumor necrosis factor-alpha (TNF-alpha) was recently reported to be elevated in acute rejection of solid organ transplants. This cytokine is released early in immune activation and may be detected in the peripheral circulation. Serial determinations of TNF-alpha serum levels were performed following experimental corneal allografts. Lewis rats received 3.5 mm orthotopic corneal grafts of MHC incompatible Wistar-Furth donors. TNF-alpha concentrations were measured in serum samples collected pre- and postoperatively and measured by micro ELISA. Clinical observations revealed graft rejection in 65.5% of corneal transplants 14 +/- 4 days following grafting. The mean serum level of TNF-alpha in control animals without corneal graft (group I) was 41 +/- 12 pg/ml. Animals following keratoplasty without allograft rejection (group II) showed a mean TNF-alpha level of 54 +/- 16 pg/ml that did not differ from group I. The rejection group III displayed significantly higher TNF-alpha levels (98 +/- 16 pg/ml, p < 0.05). These significantly elevated levels were found even before the diagnosis of rejection was established by clinical criteria. These data suggest systemic immunoreactivity to corneal allografts. Elevated levels of cytokines may provide valuable information in recipients undergoing rejection and may also provide a rationale for systemic immunotherapy in some instances. PMID- 9326760 TI - Pleomorphism of stromal eosinophils in murine experimental onchocercal keratitis. AB - Onchocercal keratitis (river blindness) is one of the leading worldwide causes of blindness. Light microscopic analysis of human specimens and corneal tissue from experimental models has implicated the eosinophil as an important cell in the inflammatory response. Our previous studies in experimental murine onchocercal keratitis have demonstrated that the inflammatory infiltrate is composed primarily of eosinophils displaying ring shaped or bilobed nuclei. However, a number of cells were not characterizable by light microscopy, presumably due to mechanical distortion. To more fully characterize the inflammatory cell infiltrate, we examined corneal specimens by transmission electron microscopy. In addition to typical eosinophils with bilobed and ring shaped nuclei, this approach revealed cells with variable nuclear morphology and cell shape which contained the dense cored granules characteristic of eosinophils. Hence, the degree of pleomorphism of eosinophils is broader than appreciated and underscores the importance of this cell in experimental murine onchocercal keratitis. PMID- 9326761 TI - Prevention of experimental autoimmune uveoretinitis by intrathymic S-antigen injection. AB - The objective of this paper was to determine whether intrathymic injection of retinal S-antigen (S-Ag) can prevent experimental autoimmune uveoretinitis (EAU) in Lewis rats. Lewis rats were injected intrathymically with 25-100 micrograms of S-Ag in 100 microliters split between thymic lobes. Controls received vehicle alone (PBS) or 100 micrograms of BSA. Animals were immunized two weeks later with 100 micrograms of S-Ag in CFA with or without pertussis toxin (0.5 micrograms/rat). Clinical ocular disease was confirmed by histopathology. Splenocytes and lymph node cells were assayed, in vitro, for their ability to proliferate in response to various concentrations of S-Ag. Furthermore, attempts were made to adoptively transfer protection to naive rats using spleen cells from intrathymically injected animals and to adoptively transfer EAU to protected rats using Con A activated cells from affected animals. Intrathymic injection of S-Ag reduced the incidence of EAU in animals subsequently immunized with S-Ag and pertussis, and prevented it entirely in rats immunized in the absence of pertussis. Splenic and lymph node cells from intrathymically injected animals showed reduced reactivity to S-Ag compared to controls, suggesting that intrathymic S-Ag injection may have rendered them tolerant to this antigen. We were unable to adoptively transfer protection to naive rats, nor were intrathymically injected rats protected from EAU induced by the adoptive transfer of primed lymph node cells. These data demonstrate that intrathymic S-Ag injection can be an effective method for protection from EAU, apparently through the induction of immunological tolerance and not active suppression. The tolerance was not absolute and could be overcome by increasing the intensity of the antigenic challenge. PMID- 9326762 TI - Tetrandrine potently inhibits herpes simplex virus type-1-induced keratitis in BALB/c mice. AB - This study investigated the effect of tetrandrine (TDR) on experimental herpes simplex keratitis (HSK) in mice. BALB/c mice were divided as follows: Group 1, untreated; Group 2, acyclovir (ACV)-treated from day 0 postinfection; Group 3, ACV-treated from day 7; Group 4, TDR-treated from day 0; and Group 5, TDR-treated from day 7. All mice were infected in the right cornea with herpes simplex virus (HSV) type I. TDR 30 mg/kg and ACV 120 mg/kg were administered intraperitoneally daily. The mice were observed for 14 days postinfection. Clinical inflammatory reactions and ocular histopathology were analyzed. The herpes specific antibody response and the delayed type hypersensitivity (DTH) response were studied. Of the 22 untreated mice, 16 developed HSK (incidence, 72.7%). TDR given from day 7 reduced the HSK incidence to 8.5% (p < 0.01); the incidence of HSK was 45.4% in mice treated with TDR from day 0 (p > 0.05). Systemic ACV given from day 0 inhibited HSK development (p < 0.01); ACV given from day 7 resulted in an HSK incidence of 50% (p > 0.05). The specific anti-HSV antibody response in the serum of mice treated with TDR or ACV either from day 0 or day 7 was significantly less than that of untreated mice (p < 0.01 and p < 0.05, respectively), and TDR treatment suppressed DTH responses to HSV (p < 0.05). Systemic TDR administered after HSV inoculation of the cornea significantly modulates murine HSK development at least partly by modifying the host immune/inflammatory response to the virus. PMID- 9326763 TI - Effect of sialodacryoadenitis virus exposure on acinar epithelial cells from the rat lacrimal gland. AB - Sialodacryoadenitis virus (SDAV), a RNA coronavirus, induces degenerative, necrotic and atrophic alterations in acinar epithelial cells of the rat lacrimal gland. To begin to explore the underlying mechanism(s) of this viral effect, we sought in the present study to: (1) determine whether SDAV invades and replicates in lacrimal gland acinar cells in vitro and (2) assess whether short-term SDAV challenge interferes with the viability or function of acinar cells in vitro. For comparison we also evaluated the relative infectivity of SDAV in acinar epithelial cells from lacrimal, submandibular and parotid glands, given that salivary tissues are known to be highly susceptible to SDAV infection in vivo. Acinar epithelial cells from lacrimal, submandibular or parotid glands were isolated from male rats, exposed briefly to SDAV or control cell antigen and then cultured for four, eight or twelve days. At experimental termination, SDAV titers in both media and sonicated cell extracts were evaluated by plaque assay titration on mouse L2 cell monolayers. To evaluate functional aspects of lacrimal gland acinar cells, SDAV-infected cells were incubated in the presence or absence of dihydrotestosterone and culture media were analyzed by RIA to measure the extent of the androgen-induced increase in secretory component (SC) production. Our results showed that: (1) SDAV invades and replicates in lacrimal gland acinar cells, Viral challenge resulted in a significant, time-dependent increase in SDAV titers, that were primarily cell-associated and greatly exceeded amounts contained in the original inoculum; (2) SDAV infection did not compromise lacrimal acinar cell viability or prevent the cellular SC response to androgens. Viral presence, though, did often attenuate the magnitude of this hormone action; and (3) SDAV infects salivary acinar cells, but the kinetics and magnitude or viral replication in lacrimal, submandibular and parotid cells showed considerable variations. These findings demonstrate that SDAV invades and replicates in acinar epithelial cells from lacrimal and salivary glands. The resulting release of infectious progeny may play a role in the SDAV-induced pathology of exocrine tissues in vivo. PMID- 9326764 TI - Pediatric uveitis in southern Turkey. AB - Uveitis in the pediatric population is not so common, and the etiology is different from adults. In the present study the charts of the patients with onset of uveitis at 16 years of age or younger were reviewed in order to analyze the etiology in pediatric patients with uveitis, and to compare the results with other studies carried out on different populations. The charts of 90 cases, followed at the Uvea-Immunology Clinic of the Cukurova University Medical Faculty Department of Ophthalmology, between January 1987 and February 1996 were reviewed retrospectively. There were 47 girls, 43 boys, aged 5-20 (13.64 +/- 4.24) in the study group. The average age at onset of uveitis was 12.20 +/- 4.81, and the follow-up period was 9-98 months (21.80 +/- 11.13). Of the 90 patients 31 (34.4%) had panuveitis, 30 (33.3%) had anterior uveitis, 21 (23.3%) had posterior uveitis and 8 (8.9%) had intermediate uveitis. Only 2 (9.5%) of the patients with posterior uveitis were considered idiopathic after extensive laboratory and clinical work-up, whereas idiopathic cases constituted 48.4% of panuveitis, and 46.7% of anterior uveitis cases. Of the 90 patients an associated condition could be found in only 59 (65.6%) patients. Of these 59 patients 23 had toxoplasmosis (39%), which constituted the most common associated condition in this study. The second most common underlying cause was Behcet's disease (17%), followed by pars planitis (13.6%), Fuchs' heterochromic iridocyclitis (8.5%), JRA (5.1%), leukemia (5.1%), and herpetic eye disease (5.1%). There were single cases with Reiter, toxocariasis, traumatic uveitis, and sympathetic ophthalmia. Environmental, cultural and genetic factors may be accountable for the differences in relative frequencies of some of the associated conditions between our findings and those of previously published studies in patients with pediatric uveitis. PMID- 9326765 TI - Prolonged retinal arterio-venous circulation time by fluorescein but not by indocyanine green angiography in birdshot chorioretinopathy. AB - PURPOSE: To compare retinal arterio-venous circulation time by fluorescein (FA) and indocyanine green angiography (ICGA) in birdshot chorioretinopathy. METHODS: We analyzed prolonged retinal arterio-venous fluorescein transit time, a known feature in birdshot chorioretinopathy and correlated it with ICGA findings in four consecutive patients. RESULTS: Mean retinal arterio-venous fluorescein circulation time was 31.1 +/- 5.2 seconds, a transit time significantly longer than in a group of ten patients with sarcoidosis (9.45 +/- 3.36 sec., p < 0.0001) and in a group of three cases with Vogt-Koyanagi-Harada disease (7.0 +/- 1.1 sec., p < 0.0001). CONCLUSION: Prolonged fluorescein arterio-venous transit time seems to be a characteristic feature of birdshot chorioretinopathy that does however not reflect the actual intravascular hemodynamic situation but diffuse blood-retinal barrier damage allowing exudation, slow gradual tissue impregnation and delayed venous reabsorption of small molecules like fluorescein. PMID- 9326767 TI - The molecular basis of anterior associated immune deviation (ACAID). PMID- 9326766 TI - Recurrent toxoplasmic retinochoroiditis. Significance of perilesional satellite dark dots seen by indocyanine green angiography. AB - PURPOSE: To suggest an explanation for the satellite dark dots seen by indocyanine green angiography (ICGA) around the main focus of a toxoplasmic retinochoroiditis. METHODS: The authors analysed the evolution of ICG satellite dark dots in two cases of recurrent toxoplasmic retinochoroiditis receiving anti toxoplasmic treatment not including corticosteroids. RESULTS: Both patients had a recurrence on the peripheral aspect of scars from previous retinochoroiditis and were treated with pyrimethamine (50 mg/day) and sulfadiazine (4 g/day) for seven weeks. Resolution of satellite ICG dark dots was observed in both cases on the follow-up ICG angiogram performed at the end of treatment. CONCLUSION: Resolution of ICG satellite dark dots after anti-toxoplasmic treatment not including corticosteroids tends to indicate that there is probably an infectious component in these hypofluorescent dots and that they probably do not represent a purely inflammatory perilesional reaction. PMID- 9326768 TI - Molecular basis of ACAID. PMID- 9326769 TI - Studies in cranial suture biology: regional dura mater determines in vitro cranial suture fusion. AB - Craniosynostosis results in alterations in craniofacial growth that create cosmetic abnormalities and functional deficits, yet the biology underlying cranial suture fusion remains unknown. The purpose of the present study was to show that regional dura mater can induce suture fusion while in an organ culture system in cranial sutures programmed to remain patient. To accomplish this, we studied mouse cranial sutures, since in this model the posterior frontal suture (analogous to the human metopic suture) fuses in both in vivo and in vitro environments while all other sutures remain patent. We demonstrated that when mouse sagittal sutures (programmed to remain patent) were rotated or translocated to overlie the posterior frontal dura then grown in organ culture systems, suture fusion occurred. Twenty-four-day-old CD-1 mice (time when the posterior frontal suture was patent) were divided into three groups of 50 (n = 165: three groups of 50 cultured and three groups of 5 uncultured controls). Group A (unrotated control group) was characterized by a strip of posterior frontal and sagittal suture with underlying dural tissue grown in organ culture systems for up to 30 days and resulted in persistent patency of the sagittal suture and fusion of the posterior frontal suture in an anterior-to-posterior direction. Group B (rotated experimental group) was characterized by 180-degree suture rotation while in vitro and resulted in patency of the posterior frontal suture over the sagittal dura and fusion of the sagittal suture over the posterior frontal dura in a posterior-to-anterior suture direction. Group C (translocated experimental group) was characterized by translocation or shifting of sutures while in vitro and resulted in patency of the posterior frontal suture over the sagittal dura and fusion of the sagittal suture over the posterior frontal dura in an anterior-to posterior suture direction. These data from the in vitro rotation and translocation experiments indicate that the "regional" posterior frontal dura determined in vitro cranial suture fusion. Molecular mechanisms behind this process are thought to involve inductive tissue interactions of the dural cells with the suture cells by means of growth factor-mediated signal pathways. PMID- 9326770 TI - Osteoblastic and osteoclastic activation in coronal sutures undergoing fusion ex vivo. AB - Numerous studies have demonstrated the importance of dura mater in the normal development and regeneration of the cranium and its sutures. The purpose of this study was to analyze the effect of dura mater on the metabolism of bone during the process of premature suture fusion. Previously, coronal sutures of fetal rats have been shown to fuse in serum-free culture after removal of their dura mater, whereas sutures of neonatal rats resist fusion even without their dura mater present. Sutures from these two distinct developmental stages were evaluated by assaying alkaline phosphatase and tartrate-resistant acid phosphatase (TRAP), marker enzymes of bone synthesis and catabolism, respectively. Coronal sutures with adjacent calvaria were dissected from fetal day 19.5 (F19) rats (n = 142) and neonatal day 1 (N1) rats (n = 42) and randomly divided into two groups each: F19 sutures with dura mater intact; F19 sutures with dura mater removed; N1 sutures with dura mater intact; and N1 sutures with dura mater removed. Calvaria were grown in serum-free medium for up to 21 days, and enzyme activities in suture regions were assayed by microanalytical techniques at different time intervals of culture. F19 sutures without dura mater exhibited significant increases in enzyme activities during days 7 to 21 of culture, whereas those without dura mater did not. N1 sutures with or without dura mater exhibited no significant changes in enzyme activities during the 14-day period of culture. The process of F19 suture fusion, occurring in the absence of dura mater, coincided with the increased activities of both alkaline phosphatase and TRAP. These cellular, enzymatic changes may have implications for the cellular events comprising craniosynostosis in vivo. PMID- 9326771 TI - Strategy of craniofacial reconstruction after resection of spheno-orbital "en plaque" meningiomas. AB - Surgical resection of spheno-orbital "en plaque" meningiomas should be as complete as possible to prevent tumor recurrence and therefore requires a bone reconstruction. We report a series of 20 patients operated on for spheno-orbital "en plaque" meningioma between 1981 and 1993. The surgical treatment included a resection of the involved dura and a wide resection of tumoral bone using a fronto-temporal craniotomy extended to the orbitozygomaticomalar bone ridge. The craniofacial reconstruction was performed in the same operative procedure using iliac bone autograft in 11 patients, internal cortical bone from the bone flap in 8 patients, and a coral graft in 1 patient. The cosmetic result was scored according to the following criteria: superior frontal paralysis, appearance of the orbitomalar bone ridge, shape of the external temporal fossa, and projection of the eyeballs. The cosmetic result was scored as excellent or good in 17 patients, average in 2 patients, and poor in 1 patient. The iliac bone autograft appeared to be the best material for craniofacial reconstruction because it could be modeled easily to the desired shape. However, the reconstruction technique was modified as necessary according to the extent of tumor removal, clinical presentation, and age of the patient. PMID- 9326772 TI - Age related changes in intracranial volume in rabbits with craniosynostosis. AB - Neurocapsular growth is highly heritable and determines neurocranial form. Although craniosynostosis alters brain growth direction, resulting in compensatory changes in the neurocranium, it is believed that such compensations occur without reduction in intracranial volume. This hypothesis was tested in a rabbit model with nonsyndromic, familial coronal suture synostosis. Skulls of 56 rabbits (20 normals, 20 with delayed onset synostosis, and 16 with complete synostosis) were scanned using three-dimensional computed tomography at 6 and 18 weeks of age. Intracranial contents were reconstructed, and indirect intracranial volume was calculated. Qualitatively, re-formations of intracranial contents from completely synostosed rabbit skulls exhibited the typical "copper beaten" morphology. Quantitatively, intracranial volume was significantly (p < 0.05) reduced in rabbit skulls with complete synostosis compared with both control rabbit skulls and rabbit skulls with delayed onset synostosis at 6 weeks by 11 percent and 14 percent, respectively). By 18 weeks, intracranial volume in rabbit skulls with synostosis was significantly (p < 0.05) reduced (by 12 percent in complete synostosis and 8 percent in delayed onset synostosis) compared with normal rabbits. Results suggest that in rabbits with uncorrected craniosynostosis, compensatory changes in the neurocranium were not adequate to allow normal expansion of the neurocapsular matrix. Further research is needed to determine whether reduction in intracranial volume was a result of neural tissue deficiency or cerebrospinal fluid (i.e., ventricular or subarachnoid) space compression in this model. PMID- 9326773 TI - Mandibular reconstruction in children with obstructive sleep apnea due to micrognathia. AB - Obstructive sleep apnea in children with micrognathia may be severe enough to necessitate permanent airway maintenance. Affected children form a small, but important, group of patients because early reconstructive surgery to the mandible may permit decannulation. A policy of early and aggressive surgical management has been pursued for the past 15 years, and an audit of outcome was undertaken to confirm whether or not this policy has been justified. The records of 25 patients so treated between 1980 and 1994 were analyzed by an independent clinician. Decannulation was achieved in 20 of the children (80 percent). The conditions that were most amenable to successful treatment (100 percent) were hemifacial microsomia, temporomandibular ankylosis, and the Pierre Robin sequence. The outcome for children with Treacher Collins syndrome and Nager syndrome was less successful (75 and 60 percent, respectively). The worst outcome occurred in a mixed group of three children with other syndromes, two of whom failed decannulation. Surgery was most successful when carried out before the age of 2 years (87 percent) and after the age of 4 years (90 percent). PMID- 9326774 TI - Bilateral buccal flaps with double opposing Z-plasty for wider palatal clefts. AB - Described is a technique that has evolved from the challenges of closure of larger cleft palate defects and that we are now using in preference over other techniques to repair a wide variety of clefts. Soft-palate closure and muscular sling reconstruction are accomplished using a modified Furlow technique. An associated cleft of the hard palate and the gaps produced by posterior displacement of the reconstructed soft palate are closed by adding tissue, buccal flaps, rather than by closure under tension or leaving residual raw surfaces. Palate lengthening is achieved both by the Z-plasty effect and by the interposition of buccal flaps between the hard and soft palate. Seventy-six palates have been repaired using this procedure. There were three postoperative fistulas. PMID- 9326775 TI - Design of the cyclops flap for chest-wall reconstruction. AB - The cyclops flap is a little-known but not forgotten alternative in chest-wall reconstruction for women. Female patients having a large, pendulous breast with a contralateral adjacent defect may be reconstructed by this technique. The flap derives its name from the repositioning of the remaining nipple to the center of the chest. The design of the flap is described. By knowing the width and length of the defect, the surgeon can design incisions on the remaining breast tissue that will allow the flap to advance easily, reliably filling the defect. The flap is an axial-pattern flap nourished by the lateral thoracic artery and the variable external mammary artery. This arterial inflow will serve the medial portion of the flap. The operation is straightforward and predictable. In using this design scheme three times in the last two years, there have been no complications. There is little discomfort after surgery. Occasionally, patients will benefit from this reconstructive option. PMID- 9326776 TI - The effects of radiation treatment after TRAM flap breast reconstruction. AB - A subgroup of mastectomy patients receives adjuvant radiation therapy after autogenous breast reconstruction for locoregional control of cancer. The effects of radiation therapy on pedicled transverse rectus abdominis musculocutaneous (TRAM) flaps were determined to evaluate complication rates and aesthetic results. Nineteen patients from 1981 to 1994 receiving radiation therapy after a pedicled TRAM flap reconstruction were compared with 108 patients who received radiation prior to reconstruction and 572 patients who underwent breast reconstruction without radiation. Retrospective reviews of patient charts were completed. Adjuvant radiation therapy was given for chest-wall recurrence in 13 of 19 patients. With a mean follow-up of 47.6 months from the time of reconstruction, 10 patients (52.6 percent) demonstrated postradiation changes in the TRAM flap reconstruction, and 6 required surgical intervention (31.6 percent). Overall complication rates were increased but were not found to be statistically significant between the radiated TRAM flap group and patients with preoperative radiation followed by TRAM flap reconstruction (31 versus 25 percent). Fibrosis was not found in patients with pre-TRAM flap radiation or in patients without radiation but was seen in 31.6 percent of patients who received radiation after the reconstruction. Fat necrosis was not significantly increased between the two groups of radiated patients. The complication rate does not change whether a patient receives radiation before or after her reconstruction; only the nature of the complication changes (fat necrosis to fibrosis). PMID- 9326777 TI - Balloon assisted endoscopic harvest of the latissimus dorsi muscle. AB - In this study, we present our experience with balloon assisted endoscopic harvest of the latissimus dorsi muscle for extremity reconstruction. The balloon performs most of the dissection under the muscle and creates the optical work space used in the endoscopic dissection. Over the course of this series the operative time has been reduced and averaged 2 hours and 44 minutes. The reconstructive goals were met in all cases. The average axillary incision length was 5.6 cm, and there were an average of 1.3 one-centimeter or smaller counter incisions. PMID- 9326778 TI - The peritoneal free flap: an anatomic study. AB - In view of a possible clinical application of an isolated microvascular peritoneal flap, an anatomic study was performed in order to determine the peritoneal vascular territory of the deep inferior epigastric artery. For this, the deep inferior epigastric artery was injected unilaterally with Araldite in 30 embalmed cadavers and bilaterally with india ink in 15 fresh cadavers. In 70 percent of the embalmed cadavers, a constant pattern of three branches from the deep inferior epigastric artery could be identified. The peritoneal vascular supply is not derived solely from these three branches but also from multiple small branches sprouting directly from the main stem of the deep inferior epigastric artery and from segmental and muscular branches. Therefore, classification of peritoneal branches arising from the deep inferior epigastric artery seems to be of little clinical importance. In all cases, the india ink injected in the deep inferior epigastric artery colored a similar territory of the parietal peritoneum. Considering the magnitude of the peritoneal vascularization by the deep inferior epigastric artery, implementation of an isolated free or pedicled peritoneal flap seems to be possible. Such a microvascular peritoneal flap vascularized by the deep inferior epigastric artery may be used, for example, for reconstruction of mucosal defects in the head and neck region. PMID- 9326779 TI - Arterial fasciocutaneous vascular territories of the lower leg. AB - The purpose of this study was to analyze the fasciocutaneous arterial circulation of the lower extremity to provide a quantitative guide to design reliable fasciocutaneous flaps. Thirty-one fresh cadaver limbs were studied using the techniques of dissection of latex injected specimens, selective ink injections, and barium latex radiographs. Fasciocutaneous perforator locations were recorded according to fascial septum of origin and distance relative to bony landmarks between the knee and the ankle. Selective ink injections of the trifurcation vessels identified four anterior tibial, three peroneal, and three posterior tibial fasciocutaneous territories. Although perforator site locations were randomly distributed along the trifurcation vessel within any vascular territory, the separate cutaneous regions that make up the fasciocutaneous territories occur in predictable locations with a measurable standard deviation. The transverse section radiographs confirmed the transverse dimensions of the vascular territories. Additionally, the summation of any two vascular territories calculated from the anatomical data conforms to the clinically observed 2.5:1 to 3:1 length-to-width ratios for fasciocutaneous flap viability as reported by Ponten and by Barclay et al. This study provides a quantitative anatomical framework using primary fasciocutaneous vascular territories to design potentially reliable fasciocutaneous flaps in the lower extremity. PMID- 9326780 TI - Posterior interosseous free flap: various types. AB - The posterior interosseous artery is located in the intermuscular septum between the extensor carpi ulnaris and extensor digiti minimi muscles. The posterior interosseous artery is anatomically united through two main anastomoses: one proximal (at the level of the distal border of the supinator muscle) and one distal (at the most distal part of the interosseous space). In the distal part, the posterior interosseous artery joins the anterior interosseous artery to form the distal anastomosis between them. The posterior interosseous flap can be widely used as a reverse flow island flap because it is perfused by anastomoses between the anterior and the posterior interosseous arteries at the level of the wrist. The flap is not reliable whenever there is injury to the distal forearm or the wrist. To circumvent this limitation and to increase the versatility of this flap, we have refined its use as a direct flow free flap. The three types of free flaps used were (1) fasciocutaneous, (2) fasciocutaneous-fascia, and (3) fascia only. Described are 23 posterior interosseous free flaps: 13 fasciocutaneous flaps, 6 fasciocutaneous-fascial flaps, and 4 fascial flaps. There were 13 sensory flaps using the posterior antebrachial cutaneous nerve. The length and external diameter of the pedicle were measured in 35 cases. The length of the pedicle was on average 3.5 cm (range, 3.0 to 4.0 cm) and the external diameter of the artery averaged 2.2 mm (range, 2.0 to 2.5 mm). The hand was the recipient in 21 patients, and the foot in 2. All 23 flaps covered the defect successfully. PMID- 9326781 TI - The "Tic-Tac-Toe" classification system for mutilating injuries of the hand. AB - Several classifications of mutilating hand injuries exist in the literature. Unfortunately, each of these provides a categorization that is arbitrarily grouped according to the part of the hand predominantly involved. It is imperative that a comprehensive classification system incorporate the degree and precise location of soft-tissue and/or bony destruction and the vascular integrity in addition to the predominantly involved part of the hand. We therefore devised a new classification system for mutilating injuries of the hand which categorizes them into seven types: (I) dorsal mutilation, (II) palmer mutilation, (III) ulnar mutilation, (IV) radial mutilation, (V) transverse amputations, (VI) degloving injuries, and (VII) combination injuries. These types are subcategorized into three subtypes: (A) soft-tissue loss, (B) bony loss, and (C) combined tissue loss. Vascular integrity is recorded with subscript notation: (0) vascularization intact or (1) devascularization. The hand is then systematically divided into nine numerical zones in "tic-tac-toe" fashion with radial, central, and ulnar columns and proximal, central, and distal rows. The "Tic-Tac-Toe" classification system allows the examining surgeon to describe precisely any mutilating injury of the hand. This system permits accurate assessment of each hand injury by assignment of the appropriate classification type, subtype, vascular status, and zone involvement. Clinical examples illustrate the user-friendliness and practicality of this new classification system. PMID- 9326782 TI - Long-term outcome of pressure sores treated with flap coverage. AB - This article provides our experience with 45 ischial sores and 24 sacral sores in 53 paraplegic patients between 1990 and 1995. Data were evaluated as to the sites of sores and types of the transferred flaps. Types of the transferred flaps were categorized into the fasciocutaneous flap and the myocutaneous or muscle flap. In the treatment of 45 ischial sores, 18 were reconstructed with the fasciocutaneous flaps and 27 with the myocutaneous or muscle flaps. In the treatment of 24 sacral sores, 23 were reconstructed with the fasciocutaneous flaps and 1 with the myocutaneous flap. The recurrence rate was analyzed by percent pressure sore free survival (%PSFS) by the Kaplan-Meier method. Overall, the ischial sores provided a higher recurrence rate than sacral sores; however, there was no significant difference in the %PSFS between the sites of sores. The group of the sores reconstructed with the fasciocutaneous flap demonstrated significant or marginally significant better results in the %PSFS (total of ischial and sacral, p = 0.0155; ischial, p = 0.0555) compared with the group of the sores reconstructed with the myocutaneous or muscle flap. These findings indicated that the use of the fasciocutaneous flap is expected to provide a better long-term result in surgical reconstruction of pressure sores than the myocutaneous or muscle flap. PMID- 9326784 TI - The protective effect of L-arginine on ischemia-reperfusion injury in rat skin flaps. AB - The objective of this study was to examine whether the administration of L arginine, a precursor of nitric oxide and substrate of nitric oxide synthase, prior to reperfusion could lead to decrease in neutrophil-mediated tissue injury and improved flap survival. Epigastric island skin flaps were elevated in 70 rats and rendered ischemic. Thirty minutes prior to reperfusion, the rats were treated with intraperitoneal saline (n = 15), L-arginine (n = 15), D-arginine (n = 15), or N omega-nitro-L-arginine methylester plus L-arginine in equimolar amounts (n = 15). Flap survival at 7 days and neutrophil counts at 24 hours were evaluated. Flap necrosis as expected in the sham group of animals (n = 10) was 0.0 percent, while the control (saline-treated) animals had 59.6 percent necrosis. Animals treated with L-arginine demonstrated a significant decrease in flap necrosis to 12.7 percent. This protective effect was almost completely negated by N omega nitrol-L-arginine methylester, which significantly increased flap necrosis to 49.3 percent and was much less pronounced with D-arginine (28.6 percent). Neutrophil counts were significantly decreased in flaps from L-arginine-treated and sham animals versus both saline and N omega-nitro-L-arginine methylester treated groups. We conclude that administration of L-arginine prior to reperfusion can significantly reduce the extent of flap necrosis and flap neutrophil counts due to ischemia-reperfusion injury. This protective effect is completely negated by nitric oxide synthase inhibition. Since L-arginine reduces the number of neutrophils within the flap and the extent of flap necrosis only in the presence of active nitric oxide synthase, we hypothesize that this protective effect of L-arginine on ischemia-reperfusion injury is secondary to a nitric oxide-mediated suppression of neutrophil-mediated injury. PMID- 9326783 TI - Arteriovenous cross-flow flap in rats: a novel skin flap. AB - Angiosomes are blocks of tissues, composed of the integument and underlying deep structures, supplied and drained by a named artery and its accompanying vein. The purpose of the current study is to describe a new principle, which allows extension of the territory of an angiosome into the adjacent angiosome, thus enabling the creation of a large skin flap (arteriovenous cross-flow flap). Epigastric skin flaps, measuring 8 x 8 cm, were raised in 30 Sprague-Dawley male rats. In group A (single-pedicle flap), the flaps were based on the epigastric artery and vein on the ipsilateral side, and the contralateral pedicle was divided. In group B (cross-flow flap), the epigastric vein on the ipsilateral side and the epigastric artery on the contralateral side of the flap were divided. In group C (skin graft), the vascular pedicles were divided bilaterally. A definitive assessment was made on the seventh day. Digital images of the flaps were analyzed using an imaging software and the areas of skin survival and necrosis were determined. Lead oxide microangiogram was performed in another set of flaps both acutely and 1 week after flap elevation. The percent survival flap area in group A was 69.94, in group B was 89.07, and in group C was 13.00. All the groups are statistically different, with a p value < 0.001. The microangiograms showed striking differences in the vascular pattern in the cross flow and the single-pedicle flaps. It is clearly demonstrated that the arteriovenous cross-flow flaps have increased survival of skin when compared with the conventional axial-pattern flaps. PMID- 9326785 TI - Costochondral graft in acute mandibular condylar fracture. PMID- 9326786 TI - Reconstruction of a large thoracoabdominal wall defect with a flow-through forearm flap and a latissimus dorsi-groin flap. AB - We report a case of a 45-year-old man with a recurrent, large, invasive dermatofibrosarcoma protuberans over the left lower chest and abdomen. Wide surgical excision of the tumor created a major thoracoabdominal wall defect. Wound coverage was achieved by using a flow-through forearm flap and an inferiorly based latissimus dorsi-groin flap. Follow-up at 1 year revealed no local recurrence or herniation. PMID- 9326787 TI - Glioma of the tongue. AB - A case of glioma of the tongue that was treated successfully by simple excision and repair is presented. It may represent neural tissue that remains in the occipital somites that differentiate into the tongue muscles. Histologically, it consists of sheets of glial tissue. The literature is reviewed, and the case is discussed. PMID- 9326788 TI - Use of bone wax as a template for intraoperative evaluation of facial defects and shaping of polyethylene implants. AB - With the advent of polymer chemistry, an increasing number of alloplastic materials are now available for use as onlay implants for reconstruction of facial bony and soft-tissue deformities. An optimal clinical result of a facial contour deformity surgery will depend not only on the choice of implant, but also on the method of giving exact shape to the implant to be used. The latter is particularly important to fit the implant into the complex configuration of a specific defect of bone and soft tissue. A template greatly enhances the accuracy of implant design. In this paper we describe a new method of fabricating polyethylene implants by using bone wax as an intraoperative template. We used this technique in four patients aged 8 to 35 years (average, 18 years) with posttraumatic and congenital facial defects without any complications. We present this method as a simple, inexpensive, and accurate alternative to the more sophisticated, but expensive and time-consuming, computer-assisted implant generation. PMID- 9326789 TI - Question mark ear: a method for repair. AB - We present a rare case of bilateral question mark ears together with our method to correct the deformity. We discuss the developmental theory of this anomaly, and review the literature. We describe our technique to correct this deformity using advancement-rotation flaps taken from both sides of the ear cleft. PMID- 9326790 TI - Browpexy: lateral orbicularis muscle fixation as an adjunct to upper blepharoplasty. AB - Lateral ptosis of the eyebrow is a major part of the complex changes that patients are seeking to improve with an upper blepharoplasty. Two hundred and eight patients have undergone a browpexy procedure at the time of blepharoplasty. This operative maneuver, which is performed through the blepharoplasty incision, prevents the brow from dropping below the superior orbital rim and creates a defined tarsal sulcus to produce a sculptured upper eyelid. PMID- 9326791 TI - The inferior retinacular lateral canthoplasty: a new technique. AB - Lateral canthoplasty is a useful method of restoring lower eyelid position and thereby protecting the ocular surfaces. The success of the lateral canthoplasty procedure depends on the proper analysis of periorbital anatomy. Newer lateral canthoplasty techniques have become progressively refined in an attempt to avoid the drawbacks and pitfalls of older procedures. We present the inferior retinacular lateral canthoplasty, developed to effectively address the problems associated with lower lid laxity and/or malposition. The inferior retinacular lateral canthoplasty is a versatile reconstructive procedure that also can be used as an adjunct to aesthetic surgery. The evolution of the inferior retinacular lateral canthoplasty from over 15 years of clinical experience is discussed. PMID- 9326792 TI - Anatomy of the nasolabial fold. AB - An anatomical evaluation involving the nasolabial fold was performed on three fresh cadavers. Both gross and histologic analyses were made. The mimetic muscles were noted to have dermal extensions to the skin overlying the nasolabial fold. The dynamics of these mimetic muscles were evaluated subsequently in the clinical setting. Marked dimples and contour irregularities could be elicited by selectively contracting these muscles. Our findings suggest that it may be possible to improve the contour over the nasolabial fold during rhytidectomy procedures by severing the dermal extensions of the mimetic muscles along the nasolabial fold. This may allow better gliding of the skin and subcutaneous tissue over the nasolabial fold, thereby resulting in a smoother crease. PMID- 9326793 TI - Secondary rhytidectomy. AB - A postoperative questionnaire was sent to all secondary rhytidectomy patients inquiring about their social and physical recovery time, complications related to either the initial or secondary surgery, and the onset of any new medical problems or the commencement of any new medications between the two surgeries. The overall satisfaction rates for both surgeries, time interval between the two operations, and their perception of the years of youthful appearance gained from either operation were also investigated. The overall satisfaction rate was slightly higher for the secondary facial rhytidectomy (4.49) than for the primary rejuvenation of the face (3.97) (p < 0.06). Patients perceived themselves as looking an average of 9.31 years younger following primary surgery, as compared to an average of 10.19 years younger following the secondary rhytidectomy (p < 0.50). The average time interval between the primary and secondary rhytidectomy surgeries was 8.48 years (range = 1 to 16 years). Twenty-nine ancillary procedures were performed during the initial rhytidectomy and 70 ancillary procedures were selected during the secondary rhytidectomy (p < 0.001). There was no statistically significant difference for the physical and social recovery time between the two procedures. Fourteen of 33 patients (42.4 percent) requiring a secondary rhytidectomy had developed a new medical problem prior to the second surgery (p < 0.001) and 19 patients (57.6 percent) were started on a new medication (p < 0.001). It was concluded from this study that the secondary rhytidectomy patients are more inclined to be satisfied (approaching statistical significance), are more likely to undergo ancillary procedures, and, being 10 years older, are more prone to have medical problems with deleterious effects on surgery and to be on medications with potential ill effects. Also, observations have been made that the previous scars pose some limitations, with the anatomical changes from the previous surgeries often requiring masterful planning and execution. Skin circulation is, in general, superior, enduring more tension. PMID- 9326794 TI - CO2 laser safety considerations in facial skin resurfacing. AB - Carbon dioxide lasers have been used increasingly in the field of aesthetic plastic surgery, specifically for facial resurfacing procedures. As many plastic surgeons are now venturing into the arena of laser surgery for the first time, it is paramount to understand basic laser safety principles to protect our patients, the operating room personnel, and the laser surgeon. This article reviews basic laser principles and practices and delineates the safety requirements needed to perform laser resurfacing using the CO2 laser system. We subjected several common objects present in the operative field during resurfacing procedures to multiple passes of both the Coherent 5000 C laser and the Laser Industries (Sharplan) model 150XJ laser Silktouch to assess flammability and margins of safety. We tested endotracheal tubes, wet and dry towels, wet and dry gauze sponges, cottonoids, eye protectors, and ophthalmic ointments. Neither flame nor burn was incited in the moistened preparations. The dry objects tested produced flame. The plastic corneal protectors began to melt by the third pass and produced significant heat. Lastly, both the Lacrilube and Bacitracin ophthalmic ointments began to vaporize after three laser passes. On the basis of our findings in this study, we recommend guidelines for prudent and safe CO2 laser usage in facial skin resurfacing. PMID- 9326795 TI - Measurement of 2,4-toluenediamine in urine and serum samples from women with Meme or Replicon breast implants. AB - The objective of this matched case-control study was to determine whether women with Meme or Replicon polyurethane-covered silicone breast implants are exposed to clinically significant levels of free 2,4-TDA from biodegradation of the polyurethane foam. Urine and serum samples were obtained from 61 patients with Meme or Replicon breast implants and 61 controls on two separate occasions separated by 10 +/- 3 days. Free TDA was analyzed by gas chromatography combined with negative chemical ionization mass spectrometry with lower limit of quantitation in both urine and serum of 10 pg/ml. The results were correlated with the length of time since implantation. No patients or controls had detectable free 2,4-TDA in their sera. Thirty patients had quantifiable levels of free 2,4-TDA, and 18 had detectable levels in their urine. Controls had no quantifiable levels, but 7 subjects had detectable levels. The biodegradative half-life of the polyurethane foam was estimated to be 2 years. A risk assessment using the cancer potency estimate calculated by the FDA from rat data and the National Academy of Sciences methodology provided a theoretical lifetime risk of approximately one in one million. It was concluded that the polyurethane foam cover on the Meme and Replicon breast implants biodegrades. The risk assessment of approximately one in one million derived from this study strengthens earlier conclusions by the Health Protection Branch (Canada) that there is no significant risk of cancer from exposure to the 2,4-TDA formed from this biodegradation. PMID- 9326796 TI - Body image and psychological sequelae of silicone breast explantation: preliminary findings. AB - Twenty-two breast explantation (implant removal) and 20 cholecystectomy patients were assessed preoperatively and postoperatively and compared with 20 nonsurgical control subjects on several body-image measures, depression, self-esteem, and self-reported health status. Explantation patients had higher breast anxiety and upper torso dissatisfaction than either control group and levels were unaffected by implant removal. The discrepancy between self-rated ideal and current breast size increased substantially after implant removal for the explantation group, but did not change for controls. Overall appearance satisfaction level and positive appearance-related cognitions decreased as a function of surgery for explantation patients, but remained unchanged in cholecystectomy and nonsurgical controls. Depression levels were elevated in explantation patients and did not change as a function of surgery; self-reported health status level improved for the explantation group, but levels still remained below those of both control groups after explantation. Therapeutic indications for the elevated depression levels and unique body-image issues that patients undergoing explantation experience are discussed. PMID- 9326797 TI - Silicone and plastic surgery. PMID- 9326798 TI - A review of epidemiologic studies analyzing the relationship between breast implants and connective tissue diseases. PMID- 9326799 TI - On the safety of breast implants. AB - A great deal of recent safety research combined with over 30 years of clinical experience has proven the value and relative safety of these devices. Aside from the side effect of capsular contracture, the complication rate of this surgery approaches that of any clean elective procedure. There is no convincing evidence to date of any systemic disorder that can be attributed to the silicone. Because these are man-made devices they do have a failure rate and in some women can require a significant amount of maintenance. As with all natural body parts these artificial substitutes may be subject to injury or disease and when viewed from that perspective do have very favorable risk benefit characteristics. PMID- 9326800 TI - Daubert and the breast implant litigation: how is the judiciary addressing the science? PMID- 9326801 TI - Use of endoscopic surgery for forehead recontouring. AB - Forehead recontouring in endoscopic surgery is presented. Eleven cases of protruded forehead deformity caused by benign tumor and one case of concave deformity caused by depressed frontal bone fracture were treated. All lesions were approached through incisions made in the hair-bearing area and operated on endoscopically. This method left no scars on the forehead, and the results were satisfactory. It is considered to be an excellent procedure with regard to cosmetic results. PMID- 9326802 TI - Graduate medical education in plastic surgery: a time for revolution. PMID- 9326803 TI - Complications and toxicities of implantable biomaterials used in facial reconstructive and aesthetic surgery: a comprehensive review of the literature. AB - The use of implantable biomaterials has become an integral part of aesthetic and reconstructive surgery of the face. Metals are used for fracture fixation devices, whereas polymers are used primarily for bone or soft-tissue substitution. This review of the scientific literature examines the risks and complications of these materials. First, we present an overview of commonly used materials. Second, we address general considerations of toxicity relevant to all biomaterials. Third, we present data from a large number of clinical series on the incidence of complications for individual materials used in specific applications. PMID- 9326804 TI - Suturing transconjunctival incision: the transpalpebral intraconjunctival suture. PMID- 9326805 TI - Herpes zoster after breast augmentation. PMID- 9326806 TI - The internal mammary artery and vein as a recipient site for free-flap breast reconstruction. PMID- 9326808 TI - Formula to estimate operating time when the procedure is done by two primary surgeons. PMID- 9326807 TI - Internal mammary artery and vein. PMID- 9326809 TI - Animal research. PMID- 9326810 TI - Peroneal functional index. PMID- 9326811 TI - Vermilionectomy and mucosal advancement. PMID- 9326812 TI - Deep vein thrombosis after face-lift surgery. PMID- 9326813 TI - Tumescent liposuction complicated by pulmonary edema. PMID- 9326814 TI - Vascular malformations and Binder's syndrome. PMID- 9326815 TI - Psychiatry of old age is coming of age. PMID- 9326816 TI - Clinical genetics, IV. Alzheimer's disease: from gene to pathology. PMID- 9326817 TI - Mortality of elderly patients with psychiatric disorders. AB - OBJECTIVE: The goal of this study was to evaluate the impact of common late-life mental disorders on the life expectancy and causes of death of older psychiatric patients. METHOD: The study population consisted of 809 older psychiatric patients who met DSM-III-R criteria for organic mental disorders, mood disorders, or psychotic disorders and who were discharged after a comprehensive multidisciplinary evaluation and acute inpatient treatment for their behavioral disorders. Dates and causes of death during a 5.75-year follow-up period were provided by the Pennsylvania Department of Health. Univariate and multivariate survival procedures were used to compare the survival rates of the three groups to each other and to a reference population of Pennsylvania residents. Causes of death were also tabulated according to ICD-9-CM and compared across the groups. RESULTS: Age, gender, race, and medical comorbidity made significant independent contributions to survival. When these variables were controlled, the survival of patients with organic mental disorders was less than half of that for patients with mood or psychotic disorders. However, all three groups experienced higher rates of mortality than the reference population, with standardized mortality ratios of 1.5 to 2.5. Deaths occurred from the usual spectrum of natural causes, with the exception that patients with mood disorders were more likely to have died from disorders of the digestive system and suicide. CONCLUSIONS: The mental disorders of late life have a significant negative impact on the survival of older psychiatric patients. PMID- 9326818 TI - Use of antidepressants by nonpsychiatrists in the treatment of medically ill hospitalized depressed elderly patients. AB - OBJECTIVE: The purpose of this study was to examine antidepressant use by nonpsychiatrists in the treatment of depressed elderly medical inpatients. METHOD: Patients aged 60 or older who were admitted to medical services at Duke Hospital were evaluated by a geropsychiatrist who used a structured psychiatric interview to identify major or minor depressive disorder. Medical records of depressed patients were reviewed for use of antidepressants and benzodiazepines before admission, during hospitalization, and on discharge. After discharge, depressed patients were contacted four times by telephone at 12-week intervals to inquire about medication use (median follow-up time = 45 weeks). RESULTS: Of 153 depressed patients, 40.5% received antidepressants at some time during their hospital stay or follow-up period, 25.5% received only benzodiazepines, and 34.0% received neither. The most commonly prescribed antidepressant was amitriptyline (45.2% of treated patients), administered at an average maximum dose of 49 mg/day. Only 15 of 114 untreated depressed patients started antidepressant therapy during hospitalization (nine with amitriptyline). Of 91 depressed patients who did not receive antidepressants either before admission or during hospitalization, only 11% received any antidepressant therapy during the median 11-month follow-up; again, half were treated with amitriptyline at doses of 10-30 mg/day. Intensity of antidepressant therapy was predicted by severity of depressive symptoms, history of psychiatric problems, and higher income. CONCLUSIONS: A relatively low proportion of depressed older medical inpatients receive treatment with antidepressants. Patients treated with antidepressants often receive potentially dangerous tertiary tricyclics at inadequate doses. Unless depression is identified and treated during medical hospitalization, it is unlikely to be treated adequately after discharge. PMID- 9326819 TI - Depression in medically ill hospitalized older adults: prevalence, characteristics, and course of symptoms according to six diagnostic schemes. AB - OBJECTIVE: The purpose of this study was to examine and compare rates of depression, correlates, and course of symptoms in medically ill hospitalized elders through use of six diagnostic schemes (inclusive, etiologic, exclusive inclusive, exclusive-etiologic, substitutive-inclusive, and substitutive etiologic). METHOD: A consecutive series of 460 cognitively unimpaired patients aged 60 or over who were admitted to the medical inpatient services of Duke Hospital underwent a structured psychiatric evaluation administered by a psychiatrist. Patients with depression were contacted by telephone at 12-week intervals after discharge to assess weekly change in depressive symptoms (median follow-up time = 47 weeks). RESULTS: The prevalence of major depression varied from 10% to 21% depending on diagnostic scheme; similarly, minor depression varied from 14% to 25%. Diagnostic strategy made little difference in known psychological and health characteristics of patients with depression (predictive validity) or severity of depressive symptoms (convergent validity). The diagnostic strategy that best distinguished a severe and persistent major depression was the exclusive-etiologic approach; however, this strategy missed 49% of patients with major depression identified by the inclusive approach, almost 60% of whom continued to experience persistent symptoms of depression many weeks after discharge. CONCLUSIONS: Diagnostic strategy affects rates of major and minor depression, with about a twofold difference between the extremes. There is little reason, however, to choose one diagnostic scheme over another in all cases. Diagnostic strategy should be chosen on the basis of the specific goals and purposes of the examiner. While the exclusive-etiologic approach identifies the most severe and persistent depressions, the inclusive approach is the most sensitive and reliable approach and is an intermediate predictor of persistent depression. PMID- 9326820 TI - Does growing old increase the risk for depression? AB - OBJECTIVE: Most research examining age as a risk factor for depression has been based on cross-sectional data. To investigate the effect of aging on rates of depression prospectively, the authors used two waves of data from a panel study of community residents 50 years old and older. METHOD: Data on symptoms of major depressive episodes were examined for the 1994 and 1995 cohorts of the Alameda County Study. The authors examined age, gender, marital status, education, financial strain, chronic medical conditions, functional impairment, cognitive problems, life events, neighborhood problems, social isolation, and social support. Depression was measured with 12 items covering DSM-IV diagnostic criteria for major depressive episodes. RESULTS: Point prevalence of major depressive episodes was 8.7% in 1994 and 9.0% in 1995. Among the subjects 60 years old and older, there was a tendency toward higher prevalence in 1995. The highest prevalence rates in 1994 and in 1995 were among those 80 years old and older. Subjects who were depressed in 1994 were at greater risk for depression in 1995. When the effects of age and other psychosocial risk factors in 1994 were controlled, there were no significant age effects on depression in 1995. Multivariate analyses demonstrated that the initial age effects were due mainly to chronic health problems and functional impairment. Gender, chronic health conditions, problems with activities of daily living, cognitive problems, neighborhood problems, and social isolation in 1994 all were significant predictors of depression in 1995. CONCLUSIONS: Healthy, normally functioning older adults are at no greater risk for depression than younger adults. What seem to be age-related effects on depression are attributable to physical health problems and related disability. PMID- 9326821 TI - Effectiveness of community-based screening for depression. AB - OBJECTIVE: The effectiveness of a voluntary depression screening program was assessed by determining 1) whether participants in the 1994 National Depression Screening Day went for recommended follow-up examinations and 2) the characteristics that differentiated those who did and did not return. METHOD: Randomly selected participants (N = 1,169) from 99 facilities completed a follow up telephone survey. RESULTS: Of 805 people for whom follow-up was recommended, 56.5% (N = 455) went for an appointment. The severity of depressive symptoms in these subjects ranged from severe (33.4%, N = 152) and marked (41.3%, N = 188) to minimal (17.1%, N = 78) and normal (8.1%, N = 37). Subjects with marked or severe depression were more likely to respond to the screening recommendation than were those with minimal depressive symptoms. However, at each level of symptom severity, subjects who had received previous treatment were more likely to adhere to the screening recommendation than were those with no previous treatment. Of those who returned for a recommended follow-up, 72.1% were diagnosed with depression. Of those who did not return, 29.5% cited lack of insurance, under insurance, or inadequate finances, and 38.0% felt they could "handle" depression on their own. CONCLUSIONS: Voluntary screening for depression is an effective way to bring certain untreated depressed individuals to treatment. Inadequate insurance and the belief that individuals can manage depression on their own continue to be barriers to seeking treatment among some depressed individuals who attend a depression screening program. PMID- 9326822 TI - Social support: a genetic-epidemiologic analysis. AB - OBJECTIVE: Social support is a widely used construct in the fields of mental health, sociology, and medicine and has typically been conceptualized as an environmental factor that influences the risk for dysfunction and disease. In this study a longitudinal twin design was used to clarify the etiology of social support. METHOD: A 16-item social support inventory was administered at personal interview to a population-based sample of female twins twice, approximately 5 years apart. A twin measurement model-which permits an estimation of the etiologic role of genetic and environmental factors correcting for errors of measurement or short-term temporal fluctuations-was applied to these data. RESULTS: Six factors, which were moderately stable over time, were found: relative problems, friend problems, relative support, confidants, friend support, and social integration. The best-fitting twin measurement models indicated that genetic factors were of substantial etiologic significance for all six social support scales. Heritabilities of the stable component of social support ranged from 43% to 75%. Familial-environmental factors contributed to twin resemblance only for relative problems and relative support. No evidence was found for significant biases in the twin method. CONCLUSIONS: Measures of social support are moderately stable over time. When short-term fluctuations are corrected for, heritable factors are of substantial etiologic importance for social support as measured at personal interview. Treating social support solely as an environmental measure is probably incorrect. Through genetically influenced traits such as temperament, individuals play a substantial role in creating their own social environments. PMID- 9326823 TI - Social consequences of psychiatric disorders, II: Teenage parenthood. AB - OBJECTIVE: The subject of this study was the relation between retrospectively reported early-onset psychiatric disorders and subsequent teenage parenthood in the general population. METHOD: The data were from 5,877 respondents aged 15-54 years in the National Comorbidity Survey, a nationally representative household survey. Information on respondents' DSM-III-R anxiety disorders, mood disorders, substance abuse disorders, and conduct disorder, age at the birth of the first child, and teenage sexual activity was collected in face-to-face interviews. RESULTS: Early-onset psychiatric disorders were associated with subsequent teenage parenthood among both females and males, with significant odds ratios of 2.0-12.0 and population attributable risk proportions of 6.2%-33.7%. Disaggregation analyses showed that disorders were associated with increased probability of sexual activity but not with decreased probability of using contraception. CONCLUSIONS: These results add to a growing body of evidence that psychiatric disorders are associated with a variety of adverse life consequences. The current policy debate concerning universal insurance coverage needs to take this into consideration. Planners of interventions aimed at preventing teenage pregnancy should consider including a mental health treatment component in their intervention packages. Mental health professionals treating adolescents need to be sensitized to their higher risk of pregnancy, while family doctors and specialists treating teenage mothers or their children need to be sensitized to the mothers' higher risk of psychiatric disorder. PMID- 9326824 TI - Clinical factors contributing to the differential diagnosis of primary insomnia and insomnia related to mental disorders. AB - OBJECTIVE: Primary insomnia and insomnia related to mental disorders are the two most common DSM-IV insomnia diagnoses, but distinguishing between them is difficult in clinical practice. This analysis was performed to identify clinical factors used by sleep specialists to distinguish primary insomnia from insomnia related to mental disorders. METHOD: Clinicians evaluated 216 patients referred for insomnia at five clinical sites, rated a list of clinical factors judged to contribute to each patient's presentation, and assigned diagnoses. Analysis of variance was performed, with contributing factors as the dependent variable and diagnostic group and clinic location as independent variables. RESULTS: Sleep specialists rated a psychiatric disorder as a stronger factor for insomnia related to mental disorders and rated negative conditioning and sleep hygiene as stronger factors for primary insomnia. However, a psychiatric disorder was rated as a contributing factor for 77% of patients who received a first diagnosis of primary insomnia. CONCLUSIONS: While neither sleep hygiene nor negative conditioning is a diagnostic criterion in DSM-IV, these results support the face validity of these clinical factors distinguishing between primary insomnia and insomnia related to mental disorders. The use of a psychiatric disorder as an inclusion criterion for insomnia related to mental disorders and an exclusion criterion for primary insomnia reinforces a categorical distinction between the two diagnoses, but the contribution of psychiatric symptoms in primary insomnia appears to be a clinically relevant one. These findings suggest the need for studies on the validity of negative conditioning and sleep hygiene in the etiology of primary insomnia, as well as on the significance of psychiatric disorders, especially depression, in primary insomnia. PMID- 9326825 TI - Prevalence, burden, and treatment of insomnia in primary care. AB - OBJECTIVE: The prevalence, burden, and management of insomnia among primary care patients were evaluated. METHOD: Consecutive patients aged 18 to 65 years in primary care clinics of a staff-model health maintenance organization (N = 1,962) were screened with the 12-item General Health Questionnaire. A stratified random sample (N = 373) completed face-to-face diagnostic assessments including the Composite International Diagnostic Interview, a brief self-rated disability questionnaire (Brief Disability Questionnaire), and the interviewer-rated Social Disability Schedule. A telephone follow-up survey was completed 3 months later. Use of psychotropic drugs, use of mental health services, and direct health care costs were assessed by using the health plan's automated data systems. RESULTS: Approximately 10% of the primary care patients reported major current insomnia (e.g., taking at least 2 hours to fall asleep nearly every night). Current insomnia was associated with significantly greater functional impairment (according to both Brief Disability Questionnaire and Social Disability Schedule), more days of disability due to health problems, and greater general medical service utilization. While insomnia was associated with depressive disorder and chronic medical illness, adjustment for these factors only partially accounted for the association of insomnia with disability and with health care utilization. Of the patients with current insomnia, 28% received any psychotropic drug; 14% received benzodiazepines and 19% received antidepressants. CONCLUSIONS: Insomnia among primary care patients is associated with greater functional impairment, lost productivity, and excess health care utilization. PMID- 9326826 TI - Small planum temporale volume in Down's syndrome: a volumetric MRI study. AB - OBJECTIVE: Down's syndrome is associated with structural brain abnormalities and language deficits. The aim of this study was to investigate whether the superior temporal gyrus and the planum temporale, both parts of the anatomic substrate for language, are abnormal in Down's syndrome. METHOD: The authors examined volumetric magnetic resonance imaging (MRI) measures of the superior temporal gyrus and the planum temporale for 17 community-dwelling patients with Down's syndrome and 17 matched healthy comparison subjects. For the subjects with Down's syndrome, the correlations of the superior temporal gyrus and planum temporale volumes with performance on tests of language function were examined. RESULTS: The planum temporale volume of the patients with Down's syndrome was smaller than that of the healthy subjects, even after differences in whole brain volume were controlled for. The volume of the superior temporal gyrus in the Down's syndrome patients was proportionally similar to that of the comparison group. For the subjects with Down's syndrome, neither superior temporal gyrus nor planum temporale volume was significantly correlated with performance on language tests after total brain volume was controlled for. CONCLUSIONS: In Down's syndrome, planum temporale volume may be selectively smaller than normal, although the effect of this volume deficit on language is not clear. PMID- 9326827 TI - Central serotonin activity and aggression: inverse relationship with prolactin response to d-fenfluramine, but not CSF 5-HIAA concentration, in human subjects. AB - OBJECTIVE: This study compared the nature and magnitude of the relationship between aggression and CSF 5-hydroxyindoleacetic acid (5-HIAA) concentration with that between aggression and the prolactin response to d-fenfluramine challenge in human subjects. METHOD: The Life History of Aggression assessment scores of 24 subjects with personality disorders were compared with their lumbar CSF 5-HIAA concentrations and with their prolactin responses to d-fenfluramine challenge. RESULTS: Aggression was significantly and inversely correlated with prolactin responses to d-fenfluramine challenge but not with lumbar CSF 5-HIAA concentrations in these subjects. CONCLUSIONS: Prolactin response to d fenfluramine may be more sensitive than lumbar CSF 5-HIAA concentration in detecting a relationship between aggression and central serotonin activity in noncriminally violent human subjects. PMID- 9326829 TI - Trichotillomania: behavioral symptom or clinical syndrome? PMID- 9326828 TI - Seasonality of symptoms in women with late luteal phase dysphoric disorder. AB - OBJECTIVE: Both late luteal phase dysphoric disorder (LLPDD) and seasonal affective disorder are cyclical disorders often manifested by "atypical" depressive features. The goal of this study was to determine whether patients with LLPDD demonstrate substantial seasonal variation in symptoms. METHOD: Consecutive female patients attending a subspecialty clinic in a university teaching hospital were assessed by means of DSM-III-R criteria. All subjects completed the Seasonal Pattern Assessment Questionnaire, modified to include items on the seasonality of premenstrual symptoms. The results were compared with those of a group of female nonclinical subjects (N = 50). RESULTS: One hundred patients met the DSM-III-R criteria for LLPDD. Compared to the nonclinical group, the LLPDD patients had a significantly higher mean global seasonality score (an index of seasonality of mood and vegetative symptoms) and a significantly higher rate of seasonal affective disorder (38% versus 8%) as determined by Seasonal Pattern Assessment Questionnaire criteria. Twenty-five percent of the LLPDD group rated their seasonal variation in premenstrual symptoms as marked or severe, while 30% considered seasonal changes in overall symptoms to be a marked or severe problem. CONCLUSIONS: These results suggest that patients with LLPDD have substantial seasonal patterns in mood and premenstrual symptoms. These seasonal patterns have implications for the clinical assessment and treatment of LLPDD. For example, light therapy may be beneficial for women with seasonal worsening of LLPDD. PMID- 9326831 TI - Possible association of a polymorphism of the tryptophan hydroxylase gene with suicidal behavior in depressed patients. AB - OBJECTIVE: This study was designed to test the hypothesis that serotonin-system related genes may be correlated with suicide risk. METHOD: Fifty-one unrelated Caucasian inpatients with major depression, with or without a history of suicidal acts, were genotyped for a biallelic polymorphism at the tryptophan hydroxylase locus. RESULTS: The less common tryptophan hydroxylase U allele occurred with greater frequency in the patients who had attempted suicide. A logistic regression analysis confirmed an association between tryptophan hydroxylase genotype and lifetime history of suicide attempts. CONCLUSIONS: Serotonergic system-related genes may influence the risk of suicide in persons with major depression. PMID- 9326832 TI - Luteinizing hormone pulse characteristics in depressed women. AB - OBJECTIVE: Luteinizing hormone (LH) pulse characteristics in depressed and normal women were compared to determine whether hypothalamic dysregulation in depression extends to the hypothalamic-pituitary-gonadal axis. METHOD: The subjects were 10 depressed and 13 normal comparison women admitted to a clinical research center. For each woman, an intravenous line was started and blood was withdrawn every 10 minutes for 8 hours. Blood samples were assayed for LH and LH pulse characteristics determined by using the computerized cluster algorithm of Veldhuis and Johnson. RESULTS: The depressed women differed significantly from the comparison women in LH pulse amplitude, rhythmicity, and area under the curve. CONCLUSIONS: Major depressive disorder is associated with abnormal regulation of luteinizing hormone. Gonadotropin regulation may provide a hormonal link between major depressive disorder and impaired fertility. PMID- 9326833 TI - Valproate prophylaxis in a prospective clinical trial of refractory bipolar disorder. AB - OBJECTIVE: The authors studied the efficacy of valproate plus lithium and of triple therapy with lithium, carbamazepine, and valproate in refractory bipolar illness. METHOD: The subjects were 24 bipolar outpatients who had completed an intended 3-year crossover study comparing lithium, carbamazepine, and their combination. Patients entered a 1-year phase of valproate plus lithium because of inadequate response or major side effects, and patients with inadequate responses were offered an additional year of treatment with all three mood-stabilizing drugs. RESULTS: Six (33%) of the 18 evaluable patients had moderate to marked responses to valproate plus lithium; four of these six had not responded to any previous treatment condition. Three of seven patients responded to triple therapy, although only one response was marked. CONCLUSIONS: Some outpatients with bipolar disorder refractory to lithium and carbamazepine received clinically relevant prophylactic benefit from valproate when used with lithium or in triple therapy. PMID- 9326834 TI - Brain proton magnetic resonance spectroscopy (1H-MRS) in Alzheimer's disease: changes after treatment with xanomeline, an M1 selective cholinergic agonist. AB - OBJECTIVE: Higher than normal cellular levels of the phospholipid catabolic intermediate glycerophosphocholine have been found in postmortem brain tissue of persons with Alzheimer's disease. Proton magnetic resonance spectroscopy (1H-MRS) can detect a choline resonance that is largely due to glycerophosphocholine. The authors tested the hypothesis that treatment with xanomeline, an M1 selective muscarinic cholinergic agonist, would be associated with a decrease in the 1H-MRS choline resonance. METHOD: Patients with mild to moderate Alzheimer's disease received placebo or xanomeline for 6 months. 1H-MRS spectra were collected at baseline and after treatment discontinuation for 12 patients, two taking placebo and 10 taking xanomeline at a dose of 25 mg t.i.d. (N = 4), 50 mg t.i.d. (N = 3), or 75 mg t.i.d. (N = 3). RESULTS: For the combined group of patients taking xanomeline, there was a significant decrease in the choline/creatine ratio from baseline to endpoint. CONCLUSIONS: Treatment of Alzheimer's disease with a cholinergic agonist is associated with a decrease in the MRS choline resonance. Xanomeline may reduce breakdown of cholinergic neuron membranes by reducing the cellular requirement for free choline for acetylcholine synthesis. PMID- 9326835 TI - False positive results: a challenge for psychiatric screening in primary care. AB - OBJECTIVE: The goal of this study was to characterize primary care patients with false positive results on screens for mental disorders. METHOD: A sample of 1,001 primary care patients completed self-administered screens and structured interviews for DSM-IV diagnoses. RESULTS: A substantial proportion of the patients with false positive screen results for at least one diagnosis met the diagnostic criteria for other psychiatric disorders. They also had significantly greater functional impairment and higher rates of recent use of mental health services than the subjects with true negative results on the screens. CONCLUSIONS: Although the positive predictive values of screens for specific mental disorders are in line with those of other medical screens, false positive results are not uncommon. This may be due in part to the sensitivity of brief screening instruments to nonspecific symptoms. The results suggest that as with other screens used in primary care, patients with false positive results on screens for mental disorders should receive clinical attention. PMID- 9326836 TI - Lethal interaction of clozapine and buspirone? PMID- 9326838 TI - Withdrawal syndrome after discontinuation of venlafaxine. PMID- 9326839 TI - Paroxetine treatment of sexual disinhibition in dementia. PMID- 9326837 TI - Clozapine-induced stuttering. PMID- 9326840 TI - Extrapyramidal symptoms from intravenous haloperidol in the treatment of delirium. PMID- 9326842 TI - Risperidone for treatment of childhood schizophrenia. PMID- 9326841 TI - Atypical neuroleptic malignant syndrome associated with risperidone treatment. PMID- 9326843 TI - E-mail therapy. PMID- 9326844 TI - Lithium assay errors. PMID- 9326845 TI - Uncontrolled buying. PMID- 9326846 TI - Self-reported life satisfaction. PMID- 9326847 TI - Self-reported life satisfaction. PMID- 9326848 TI - Effects of pregnancy on suicidal behavior. PMID- 9326849 TI - Benzodiazepine receptor binding and schizophrenia. PMID- 9326850 TI - Violence and cocaine. PMID- 9326851 TI - Psychotherapy's role in psychiatry. PMID- 9326852 TI - Peacekeeping duty and PTSD. PMID- 9326853 TI - Confidentiality conundrum. PMID- 9326855 TI - Borderline personality disorder and transitional objects. PMID- 9326854 TI - Genetic risk factors for bipolar disorder. PMID- 9326856 TI - Italian psychiatric reform. PMID- 9326857 TI - Italian psychiatric reform. PMID- 9326858 TI - Recognition of depression in geriatric ED patients by emergency physicians. AB - STUDY OBJECTIVE: To prospectively evaluate identification of geriatric depression by emergency physicians and to assess the utility of a self-rated depression scale to improve case-finding in geriatric patients presenting to the ED. METHODS: We conducted an observational survey of geriatric ED patients who presented to an urban, university-affiliated public hospital. A brief self-rated depression scale was administered to 101 patients aged 65 years or older. Emergency physicians, blinded to depression scale scores, prospectively rated the likelihood of depression in these patients. Our main outcome measures were prevalence of depression (in accordance with a predetermined cutoff score for detecting depression) and the emergency physicians' clinical recognition of depression. RESULTS: Thirty patients (30%; 95% confidence interval [CI], 21% to 39%) met the predetermined criteria for depression. Age, sex, race, and education were not significantly different between depressed and nondepressed patients. Patients who categorized their health as good were less likely to be depressed than those who considered their health poor or fair (18% versus 37%; 95% CI for difference of 19%, 10% to 35%). Recognition of depression by emergency physicians was poor, with a sensitivity of 27% (95% CI; 12% to 46%), specificity of 75% (95% CI, 63% to 84%), and positive predictive value of 32% (95% CI, 27% to 41%). Only 13% (95% CI, 4% to 31%) of depressed patients were referred for further mental health evaluation. CONCLUSION: Depression is common in older ED patients but often goes unrecognized by emergency physicians. Use of a brief depression scale can improve case-finding in this age group, leading to appropriate referral for further management. PMID- 9326859 TI - ED use by older victims of family violence. AB - STUDY OBJECTIVE: To determine the nature and frequency of ED use by victims of physical elder abuse. METHODS: Community-dwelling victims of abuse were identified through a state elderly protective service program independent of the health care system in a geographic area served by two EDs. ED records were reviewed and abstracted to determine if and how victims used emergency services. RESULTS: During a 7-year period, 182 elderly victims of physical abuse were identified in the catchment area of the study, and 114 (62.6%) had been seen in one or both EDs at least once during a 5-year "window" surrounding the initial identification of abuse. These 114 individuals accounted for 628 visits (median 3, range 1-46); 30.6% of visits resulted in a hospital admission. An ordinal system was used that assigned a probability of any single ED visit being referable to abuse; 37.8% of subjects had at least one visit categorized as being of high probability, and 66% of subjects had at least one visit that resulted in an injury-related chief symptom or ICD-9 discharge diagnosis. CONCLUSION: Elder abuse victims have substantial interactions with EDs and these visits frequently result in admission. Strategies that identify elder abuse in less acute settings and effectively address the needs of victims would improve quality of life and likely result in substantial savings in health care expenditures. PMID- 9326860 TI - Immunologic response to tetanus toxoid in geriatric patients. AB - STUDY OBJECTIVE: Tetanus antibody levels have been shown to be inadequate in 50% of patients older than 65 years. Although immunization recommendations have been made for this age group, the efficacy of this intervention has not been well documented. We sought to determine the difference in tetanus antibody levels after the administration of one tetanus toxoid immunization to geriatric patients without adequate titers. METHODS: Thirty-five patients older than 65 years at a large urban comprehensive care geriatric center who were documented to have inadequate tetanus antibody titers were each given one tetanus toxoid immunization. Repeat titers were obtained at least 2 months after the immunization with the use of enzyme-linked immunosorbent assay (Bindazyme kit; the Binding Site Corporation, Birmingham, England). We considered tetanus antibody levels greater than .17 IU/mL protective. RESULTS: The mean age was 79.4 years; 30 of 35 (86%) were female. Repeat tetanus antibody titers were obtained an average of 123 days (range, 63 to 204 days) after immunization with tetanus toxoid. The mean preimmunization antibody titer was .1 IU/mL (range, .04 to .16 IU/mL). After immunization, antibody titers increased a mean of .61 IU/mL (range, -.01 to 2.23 IU/mL; 95% confidence interval, .35 to .87 IU/mL). Thirty of the 35 patients who received a single injection of tetanus toxoid (86%) developed protective titers. We found no relationship between seroconversion and age, sex, or medical history; nor did we find a relationship between antibody level and time elapsed since immunization when repeat titers were obtained. CONCLUSION: Administration of one tetanus toxoid injection affords protective immunity in many geriatric patients. PMID- 9326861 TI - Elder abuse: a review. AB - Elder abuse exists in many forms: physical, emotional, financial, and sexual; neglect; and self-neglect. As many as 2.5 million older people are abused each year, and the number of cases will likely increase as this population grows. Elder abuse receives less attention than other forms of domestic violence, and fewer than 10% of cases are reported. Although all states have legislation addressing elder abuse, financial support for evaluation and protective services is lacking. Most states have mandatory reporting; however, it may infringe on the autonomy of competent geriatric individuals. Physicians infrequently report elder abuse because they are not familiar with reporting laws, fear offending patients, are concerned with time limitations, and believe they do not have appropriate evaluation skills. Victims often have low self-esteem, blame themselves for the abuse, and do not want to admit their vulnerabilities or betray their families. The "caregiver stress hypothesis," which suggests abuse stems from caregiver stress and resentment resulting from chronic care of dependent geriatric patients, is a misconception. Abuse is actually better correlated with the emotional and financial dependence of the caregivers on the geriatric victims. Older patients are most commonly abused by the people with whom they live. Older men and women have similar per capita abuse rates. Assessment and management should be supportive without assigning blame and should focus on both the patient and the caregiver. Patients in immediate danger should be hospitalized or placed in emergency shelters. Suspected abuse should be reported directly to the appropriate state agency, which can provide a thorough long-term assessment. PMID- 9326862 TI - Elder mistreatment: national survey of emergency physicians. AB - To determine the perceived magnitude of elder mistreatment, physician awareness of applicable state laws, and the barriers to reporting suspected cases, we surveyed a random sample of 3,000 members of the American College of Emergency Physicians in the United States. Survey questions included practice characteristics, number and type of suspected cases of elder mistreatment seen in the ED, number of cases actually reported, and reasons for not reporting abuse. Physicians were also asked about the availability of elder-mistreatment protocols and their familiarity with local laws and reporting requirements. We received 705 completed surveys, for a response rate of 24%. Most physicians (52%) described elder mistreatment as prevalent but less so than spouse or child abuse. The respondents had evaluated a mean of 4 +/- 8 (range, 0 to 93) suspected cases of elder mistreatment in the preceding 12 months; approximately 50% were reported. Only 31% of emergency physicians reported having a written protocol for the reporting of elder mistreatment, and physicians were generally not familiar with applicable state laws. Twenty-five percent were able to recall educational content pertaining to elder mistreatment during their emergency medicine residencies. Most physicians were not certain or did not believe that clear-cut medical definitions of elder abuse or neglect exist (74%); that emergency physicians can accurately identify cases of mistreatment (58%); or that their states had sufficient resources to meet the needs of victims (92%). These results suggest that practicing emergency physicians are not confident in identifying or reporting geriatric victims of abuse or neglect. This lack of confidence may reflect inadequacies of training, research, and continuing education with regard to mistreatment of older people. PMID- 9326863 TI - Practice guideline for the ED management of falls in community-dwelling elderly persons. Kaiser Permanente Medical Group. PMID- 9326864 TI - Intravenous nitrates in the prehospital management of acute pulmonary edema. AB - STUDY OBJECTIVE: We sought to assess the effect of nitrates on prehospital mortality among patients with acute pulmonary edema (APE). METHODS: The study involved a retrospective evaluation of the records of prehospital outcome in 640 patients with APE rescued by the mobile CCU (MCCU) of Florence, Italy, between January 1980 and December 1991. The MCCU serves an urban environment with a population of 400,000 in a 102-sq km area. In the years 1980 through 1983, patients were treated with oxygen, morphine, furosemide, digoxin, nitrates, aminophylline, or dopamine, according to the attending physician's judgment. From 1984 through 1991, new guidelines for the use of intravenous nitrates, based on differential treatment according to blood pressure, were in use. RESULTS: Overall prehospital mortality rate for APE in all patients was 7.8% (50 of of 640 patients). Mortality after 1984 was significantly lower than before (5.3% versus 13%, P < .01). Nitrates were effective in reducing mortality, even in hypotensive patients. Multivariate analysis showed that outcome was significantly affected by two clinical features (dyspnea and low blood pressure), treatment with nitrates, and calendar period effects (before/after 1984). CONCLUSION: Our findings suggest that the use of intravenous nitrates improves short-term prognosis in APE. PMID- 9326865 TI - Cost-effectiveness analysis of helicopter EMS for trauma patients. AB - STUDY OBJECTIVE: To evaluate the cost-effectiveness of helicopter EMS for trauma patients. METHODS: We applied a cost-effectiveness analysis from the service provider's perspective to cost and effectiveness estimates. The cost estimates comprise direct operating costs and additional survivors' hospital costs. The effectiveness estimates were calculated with the TRISS methodology from literature sources and data from a cohort of patients transported by helicopter during 1994 and 1995. Sensitivity analysis and discounting were used. Cost per life saved and discounted cost per year of life in 1995 US dollars were the main outcome measures. RESULTS: The reported literature survival benefit ranges from 1 to 12 additional survivors per 100 patients flown. Transport costs were $2,214 per patient, and each additional survivor's hospitalization averaged $15,883. For the base case (5 additional survivors per 100 patients flown), cost per life was $60,163 and discounted cost per year of life $2,454. Sensitivity analysis revealed that discounted cost per year of life could be as high as $9,677 or as low as $1,400 and that it was most dependent on the surviving benefit. These results are comparable to a reported median discounted cost per year of life of %19,000 for other commonly used lifesaving medical interventions. CONCLUSION: Assuming that helicopter air medical transport provides a substantial survival benefit for trauma patients, our findings suggest that this service is a cost effective option for the treatment of trauma patients. The magnitude of the survival benefit is the most important factor determining cost-effectiveness. PMID- 9326866 TI - Crush syndrome sustained in the 1995 Kobe, Japan, earthquake; treatment and outcome. AB - STUDY OBJECTIVE: To assess the treatment and outcome of patients with crush syndrome sustained in an earthquake disaster. METHODS: We conducted a retrospective analysis of eight patients with crush syndrome and subsequent acute kidney failure who were treated in the ICU of a university hospital. All eight patients had been extricated from buildings that collapsed in the 1995 Kobe, Japan, earthquake. Crush injury involved the upper extremities in one patient and the lower extremities in seven. Each patient received intravenous fluid infusion and diuretic drugs and underwent hemodialysis. Emergency fasciotomy was performed in some patients, 17 to 100 hours after extrication. RESULTS: All patients were conscious and lucid on admission, and blood pressure and heart rate were normal. All the patients demonstrated kidney failure with increased concentrations of serum creatinine (1.9 to 9.6 mg/dL [169 to 852 mumol/L]). Six patients were oliguric. Hyperkalemia (5.6 to 8.8 mEq/L) was present in six patients. We found close correlations between the serum potassium and creatine kinase concentrations, between the serum myoglobin and potassium concentrations, and between the serum myoglobin and creatine kinase concentrations. All the patients were weaned from hemodialysis. The serum creatinine concentration decreased to a normal level within 20 to 52 days of admission in all patients. No patients underwent amputation. Muscle weakness and sensory deficits persisted in all patients 6 months after the earthquake. CONCLUSION: Our findings support current therapeutic strategies for crush syndrome, despite the long delay to initiation of intensive therapy. All the patients recovered kidney function and were weaned from hemodialysis; none required amputation. PMID- 9326867 TI - Emergency preparedness and response in Israel during the Gulf War. AB - We examined the effect of the emergency response on medical and public health problems during the 1991 Gulf War in Israel. On the first day of the conflict, the number of deaths from suffocation, asphyxiation, aspiration, myocardial infarction, cardiac arrest, and cerebrovascular accident increased abruptly, as did the number of sudden deaths associated with the use of tight-fitting masks with filters in sealed rooms. Much of the excess risk for death from cardiorespiratory complications during the first alert may have been a consequence of its duration (140 minutes). Mass evacuation and concrete buildings are believed to have kept the death toll from trauma down, and mask use may have protected against facial and upper-airway injuries. Falls and hip fractures, airway irritation from exposure to bleach, carbon monoxide intoxication from open kerosene heaters in sealed rooms, and self-injection with atropine syringes were also noted. A measles epidemic and increased death rates from automobile crashes were other preventable causes of death. Protection against biological warfare was limited to surveillance of trends for pneumonia and gastroenteritis. Emergency planners failed to anticipate the need for better mask fit, hands-on training in the use of masks, and special guidelines for older persons to prevent deaths from suffocation and other cardiovascular-respiratory problems in the first minutes of use. If masks are to be distributed as a protection against chemical warfare, a simpler model including the use of shrouds for whole-body skin protection might help avoid cardiorespiratory complications. Public health problems not adequately dealt with in the predisaster period are apt to emerge with greater severity during a crisis. PMID- 9326868 TI - ED evaluation of transient global amnesia. AB - Transient global amnesia is an unusual syndrome involving a transient memory loss and inability to form new memories, without affecting other cognitive functions. Although the patient often lacks insight into the problem, the syndrome is very frightening for family members, who frequently present to the ED. The evaluation of the syndrome is the subject of some controversy. This review presents current understanding about the causes of transient global amnesia and an approach to the ED evaluation of the syndrome. PMID- 9326869 TI - Emergency medicine in Bosnia and Herzegovina. PMID- 9326870 TI - Sports-related pneumothorax. AB - Pneumothorax and pneumomediastinum are rare complications of athletic activity. Spontaneous pneumothorax has been reported in association with several sports, but reports of pneumothorax associated with blunt trauma sustained during sporting activity are rare. We present a case series of patients in whom pneumothorax or pneumomediastinum developed as a result of blunt trauma sustained during participation in a contact sport. PMID- 9326871 TI - Clarithromycin-induced ventricular tachycardia. AB - Clarithromycin is a relatively new macrolide antibiotic that offers twice-daily dosing. It differs from erythromycin only in the methylation of the hydroxyl group at position 6. Although the side-effect profile of erythromycin is established, including gastroenteritis and interactions with other drugs subject to hepatic mixed-function oxidase metabolism, experience with the newer macrolides is still being recorded. Cardiotoxicity has been demonstrated after both intravenous and oral administration of erythromycin but has never been reported with the newer macrolides. We report a case of ventricular dysrhythmias that occurred after six therapeutic doses of clarithromycin. The dysrhythmias resolved after discontinuation of the drug. PMID- 9326872 TI - Pediatric duodenal perforation missed on computed tomography. AB - We present the case of a 2-year-old child who sustained duodenal perforation after blunt abdominal trauma. The presentation of gastrointestinal perforation after blunt abdominal trauma in children can be subtle and at times delayed. A slight increase in the concentration of aspartate aminotransferase, alanine aminotransferase, or pancreatic enzymes may be the only indication of hollow viscus injury. Initial radiographic studies have a low yield in detecting such injuries. Computed tomography is also unreliable. Conservative management with serial examinations and serial laboratory studies may be required to identify gastrointestinal perforation. PMID- 9326873 TI - Toxic effects of nefazodone. PMID- 9326874 TI - Toxic effects of nefazodone. PMID- 9326875 TI - Motorcycle helmets and vision. PMID- 9326876 TI - Removing inhaled teeth. PMID- 9326877 TI - Certification in emergency medicine. American College of Emergency Physicians. PMID- 9326878 TI - Emergency physician overhead. American College of Emergency Physicians. PMID- 9326879 TI - Mandatory reporting of domestic violence to law enforcement and criminal justice agencies. American College of Emergency Physicians. PMID- 9326880 TI - Military emergency medical services systems. American College of Emergency Physicians. PMID- 9326881 TI - Military emergency medicine. American College of Emergency Physicians. PMID- 9326882 TI - Emergency physician stewardship of finite resources. American College of Emergency Physicians. PMID- 9326883 TI - Schistosoma japonicum: in vitro cultivation of miracidium to daughter sporocyst using a Biomphalaria glabrata embryonic cell line. AB - In vitro cultivation of Schistosoma japonicum miracidia to the mother sporocyst (MS) and then to the daughter sporocyst (DS) stage was achieved using the Biomphalaria glabrata embryonic (Bge) cell line as a coculture system. When comparing the effect of Bge cell and MS density on MS development, it was apparent that Bge cell density had a highly significant effect on both MS viability and growth. Viability and growth rate of MS cultured under high cell density conditions (350 cells/mm2) were almost 2 times greater than those of MS cultured under conditions of low cell density (60 cells/mm2). Growth under high cell density conditions corresponded to a 20 to 30 times increase in MS estimated volume within the first 9 weeks of cultivation. Emergence of fully formed motile DS was first observed after 11 weeks of cocultivation. A few DS lived for 14 weeks after emergence and attained a size of 770 +/- 100 microns in length and 48 +/- 13 microns in width. In contrast to what was observed in Bge cell/Schistosoma mansoni cocultures, Bge cells did not encapsulate S. japonicum MS. Our results show that, although the cellular interactions between Bge cells and schistosomes MS display some level of specificity, Bge cells apparently secrete soluble factors that permit excellent survival and can trigger advanced in vitro development of S. japonicum. PMID- 9326884 TI - Toxoplasma gondii: dithiol-induced Ca2+ flux causes egress of parasites from the parasitophorous vacuole. AB - Ca2+ is an essential activator of motility in the obligate intracellular parasite Toxoplasma gondii. Ca2+ ionophore A23187 and intracellular microinjection of Ca2+ initiate motility of parasites residing in parasitophorous vacuoles (PV). The source of Ca2+ and the mechanism by which it activates motility in vivo remain uncertain. Exposure of the parasites to dithiothreitol (DTT) can activate egress of previously nonmotile intravacuolar parasites within 60 sec. DTT is also known to activate both isoforms of the highly concentrated nucleoside triphosphate hydrolase (NTPase) produced by T. gondii. Using an adherent cell analysis system (ACAS) for Ca2+ imaging, a brief 15-50% increase in intra-PV fluorescence ratio was observed after exposure of infected fibroblasts to 5 mM DTT. Chelation of intracellular Ca2+ with BAPTA-AM and extracellular Ca2+ with EGTA blocked the DTT effect; however, this chelation did not prevent the activation of parasites nor the Ca2+ response to the Ca2+ ionophore ionomycin, suggesting that the Ca2+ that activates motility may reside near or within the parasite itself. This result demonstrates that an increase in Ca2+ within the vacuole precedes the onset of motility and the correlation of the DTT effect on motility and tachyzoite NTPase suggests that NTPase activation may be involved in the Ca2+ flux. PMID- 9326885 TI - Leishmania spp.: mechanisms of toxicity of nitrogen oxidation products. AB - Intracellular killing of Leishmania parasites within activated murine macrophages is thought to result from the toxic activities of nitrogen oxidation products (referred to as NO) released by the activated cells. In order to determine possible mechanisms of NO toxicity for these microorganisms, promastigotes of Leishmania major and Leishmania enriettii were exposed to NO generated chemically from acidified nitrite, S-nitrosocysteine, diethylamine NONOate, or nitroprusside. Treatment with these agents led to loss of viability (as determined from decreased motility and inhibition of [3H]TdR uptake upon reincubation in NO-free medium) with kinetics characteristic for each compound L. major was less sensitive to these effects than L. enriettii, and amastigotes displayed the same sensitivity as promastigotes of the same species. The early effects of NO toxicity could be detected within minutes of exposure to the NO donors; they included decreased respiration rate and inhibition of glucose, proline, and adenine incorporation. Inhibition of the activities of glyceraldehyde 3-phosphate dehydrogenase and of aconitase were also evidenced. In order to determine whether these phenomena reflected the mechanisms of toxicity of bona fide NO generated by macrophages, promastigotes were exposed to IFN-gamma + LPS-activated macrophages across permeable membranes. This resulted in marked inhibition of proline and adenine uptake in the parasites, which was restored, however, to control levels when macrophages were activated in the presence of the nitric oxide synthase inhibitor NGMMA. These results indicate that several cellular targets may be subject to NO toxicity in Leishmania parasites, including enzymes of glycolysis and respiratory metabolism as well as trans-membrane transport systems. PMID- 9326886 TI - Plasmodium falciparum: cyanide-resistant oxygen consumption. AB - It has been hypothesized that Plasmodium parasites utilize a branched chain respiratory pathway, consisting of a classical cyanide-sensitive branch and an alternative cyanide-resistant branch. To further explore this hypothesis, the effect of cyanide on Plasmodium falciparum was determined using a polarographic assay. The rate of oxygen consumption by saponin-freed parasites was approximately 5% that of control human white blood cells or of Toxoplasma gondii, consistent with an anabolic role for P. falciparum respiration. However, while all of the oxygen consumption of the control white blood cells and of T. gondii could be inhibited by cyanide, 25% of the oxygen consumption of the P. falciparum parasites was found to be insensitive to high concentrations of cyanide. The cyanide-resistant portion of the parasite oxygen consumption was completely inhibited by two inhibitors of alternative oxidase activities in other systems, propyl gallate and salicyclhydroxamic acid. These studies provide the first direct evidence for a branched chain respiratory pathway in P. falciparum. Furthermore, salicyclhydroxamic acid, propyl gallate, and related inhibitors of alternative oxidase activities were shown to inhibit the growth of P. falciparum in vitro. These results support the need for further investigation of alternative oxidase activity as an antimalarial chemotherapeutic target. PMID- 9326887 TI - A novel phospholipase A2 activity in saliva of the lone star tick, Amblyomma americanum (L.). AB - Saliva from female lone star ticks, Amblyomma americanum, contained a novel phospholipase A2 (PLA2) activity that hydrolyzed 14C-arachidonate from 14C arachidonyl phosphatidylcholine. The tick saliva PLA2 (ts-PLA2) was active over a broad pH range (4.5-11.5) with two distinct pH optima of pH 5.5 and 9.5. Though extracellular PLA2s are reported to be activated by millimolar Ca2+, ts-PLA2 was sensitive to submicromolar Ca2+ and was half-maximally activated by 3.5 microM Ca2+. Tick saliva contains > 500 microM Ca2+ and the feeding lesion in the host is expected to contain millimolar Ca2+. Saliva exhibited a single peak of PLA2 activity corresponding to a molecular weight of 55.7 +/- 1.3 kDa by size exclusion chromatography. The ts-PLA2 was unaffected by a variety of compounds known to inhibit either secreted or cytosolic PLA2s from other sources. However, ts-PLA2 was inhibited by the substrate analog, oleyloxyethyl phosphorylcholine (IC50 = 1.4 microM), and the end product, arachidonic acid (IC50 = 38 microM). Low concentrations of dithiothreitol did not greatly affect ts-PLA2, but activity was reduced at higher concentrations. The PLA2 activity found in A. americanum salivary glands showed many similarities to ts-PLA2, but also some distinct differences. Secreted at the tick-host interface, ts-PLA2 is thought to play an important, but unknown, role during the prolonged tick feeding. PMID- 9326888 TI - Giardia lamblia: evidence for carrier-mediated uptake and release of conjugated bile acids. AB - Giardia lamblia trophozoites colonize the human small intestine, where they are exposed to high concentrations of conjugated bile acids. Previous work has shown that bile acids enhance trophozoite survival, multiplication, and differentiation into the cyst stage. Therefore, experiments were performed to test whether carrier-mediated uptake of conjugated bile acids is present in this primitive parasite. Uptake of both cholyltaurine (C-tau) and cholylglycine (C-gly) was increased manyfold after culturing trophozoites in medium lacking bile acids. Absence of uptake at 4 degrees C and inhibition by other conjugated bile acids provided additional evidence for carrier-mediated uptake. Uptake of C-tau was greater than that of C-gly under all experimental conditions and appeared to be mediated by a different carrier. The major evidence for different carriers is that C-tau uptake was Na(+)-dependent, while C-gly uptake was not. In addition, C tau uptake was more strongly inhibited by DTNB and several organic anions than C gly uptake. Radiolabeled C-tau and C-gly were each released rapidly from trophozoites at 37 degrees C but not at 4 degrees C, suggesting that release of conjugated bile acids was also carrier-mediated. These findings are consistent with the notion that multiple transporters for conjugated bile acids are present in a lower eukaryote. We speculate that intracellular bile acids may facilitate lipid trafficking and membrane biosynthesis. PMID- 9326889 TI - Echinococcus multilocularis metacestodes: immunological and immunocytochemical analysis of the relationships between alkaline phosphatase and the Em2 antigen. AB - Echinococcus multilocularis metacestodes possess an alkaline phosphatase (EmAP) which has been extensively characterized at the biochemical level in previous studies. The apparent molecular weight of the enzyme monomer and its isoelectric point matched those originally described for the Em2 antigen, a reference antigen currently used for the immunodiagnosis of E. multilocularis infection. These observations raised questions about the molecular relationship between the two molecules. In order to investigate the relations between EmAP and the Em2 antigen, immunoblotting and ELISA were carried out using polyclonal and monoclonal antibodies directed against EmAP and the Em2 antigen, respectively. In addition, the localization of EmAP and the Em2 antigen was compared by immunofluorescence and immunogold electron microscopy in in vitro-generated E. multilocularis metacestodes. The results show that common epitopes between EmAP and Em2 exist, which are predominantly of a peptidic nature. Both antigens are localized in an acellular parasite structure, the laminated layer, with additional locations for the EmAP on the glycocalyx and in the central region of invaginated protoscoleces. These results suggest a putative functional relationship between the two antigens and that Em2 could originate from EmAP. PMID- 9326890 TI - Plasmodium reichenowi: deduced amino acid sequence of sexual stage-specific surface antigen Prs48/45 and comparison with its homologue in Plasmodium falciparum. PMID- 9326891 TI - Pyruvate kinase is present in Giardia intestinalis. PMID- 9326892 TI - An estimation of the size distribution of amalgam particles in dental treatment waste. PMID- 9326893 TI - Dr. Basil Bibby: early fluoride investigator and intellectual provocateur. AB - Dr. Basil Bibby assumed many roles during his productive career as a researcher, teacher, and administrator. Although best known for his research on oral microbiology and on foodstuff as it relates to dental caries, and for fostering the careers of many distinguished researchers, he played an important generative role in determining the local cariostatic effects of fluoride. His seminal work in this area has not received its due because of the mixed success of his initial clinical studies. We hope that a review of his contributions in this area will bring those contributions into focus and illustrate his open personality and multi-dimensional approach to research. PMID- 9326894 TI - The activin-binding protein follistatin is expressed in developing murine molar and induces odontoblast-like cell differentiation in vitro. AB - It has recently been shown that mice deficient in activin-beta A subunits and follistatin exhibit major defects in dentition. To increase understanding of the roles played by these molecules during tooth development, we determined the temporospatial expression of activin-beta A subunit and follistatin messenger RNA and their corresponding proteins in developing murine molars (between day E 14 and 2 days after birth). The effects of recombinant human activin A and its binding protein follistatin on odontoblast differentiation were also studied in cultures of dental papillae (DP) isolated from the mandibular first molars of E 17-day mice. In situ hybridization indicated that transcripts for activin-beta A subunit were abundant in pre-odontoblasts at the tips of forming cusps prior to odontoblast terminal differentiation, and transcripts for follistatin in overlying inner enamel epithelial cells (pre-ameloblasts). Pre-odontoblasts were also weakly immunoreactive in relation to activin-beta A subunit, pre-ameloblasts in relation to follistatin. When follistatin was added at different concentrations to a DP culture model (2-14 nmol/DP) together with heparin at constant concentration, differentiation of odontoblast-like cells was induced, as evidenced by polarization and deposition of extracellular matrix in vitro, to extents depending on the follistatin concentration. In contrast, the addition of activin A (2 nmol/DP) had no effect on the differentiation parameters studied. These findings suggest that the activin-follistatin system regulates odontoblast differentiation during tooth development. In particular, we suggest that binding of endogenous activin A by follistatin may allow odontoblast terminal differentiation to occur. PMID- 9326895 TI - Salivary cholesterol of healthy adults in relation to serum cholesterol concentration and oral health. AB - Salivary lipids are mostly glandular in origin, but some are believed to diffuse directly from serum. This diffusion and the role of salivary lipids in oral health have scarcely been studied. Therefore, the serum and saliva cholesterol concentrations and oral health were analyzed in a group of healthy adults (n = 139; 64 men and 75 women; 34.2 +/- 5.2 yrs). Paraffin-stimulated whole saliva was collected, centrifuged (10,000 x g; 30 min, 4 degrees C), and lyophilized, and the cholesterol and other neutral lipids were extracted, separated by thin-layer chromatography, and quantified. The mean +/- SD (range) of saliva cholesterol concentration was 1.20 +/- 0.75 (0.02-5.46) mumol/L, and the saliva cholesterol level of men (1.36 +/- 0.85 mumol/L) was significantly higher than that of women (1.06 +/- 0.64 mumol/L; p < 0.05). Weak positive correlations between saliva and serum cholesterol concentrations and saliva cholesterol and serum non-high density lipoprotein cholesterol concentrations were found (r = 0.22, p < 0.05; r = 0.28, p < 0.005, respectively). The saliva cholesterol assay detected subjects with high (> or = 6.5 mmol/L) serum cholesterol values, with sensitivity and specificity values of 100% and 29%, respectively. A positive correlation between the body mass index and the level of saliva cholesterol concentration was also found (r = 0.31 p < 0.01). Oral health, microbial counts, or saliva flow rate revealed no differences in subjects with low and high salivary cholesterol level. We conclude that, in healthy adults, saliva cholesterol concentration reflects serum concentration to some extent and can be used to select individuals with high serum cholesterol levels. PMID- 9326896 TI - HIV status and the risk of post-extraction complications. AB - Tooth extraction is commonly performed for patients infected with the human immunodeficiency virus (HIV). We undertook a prospective study to determine if HIV-positive patients had an increased risk for complications following tooth extraction. The study sample was composed of patients who presented for tooth extraction to the outpatient oral/maxillofacial surgery clinic at Grady Memorial Hospital, Atlanta, GA. The predictor variable was HIV status (positive or negative). The outcome variable was the presence or absence of a post-extraction complication. Other study variables were grouped into the following sets: (1) demographic, (2) past medical and social history, (3) clinical, (4) laboratory values, and (5) treatment. Between 11/93 and 4/96, 166 patients were enrolled. The study sample was composed of the 151 patients who completed the study protocol and consisted of 76 HIV-positive and 75 HIV-negative patients. The post extraction complication rates were 22.3 and 13.3% for the HIV-positive and negative groups, respectively (relative risk = 1.68, 95% confidence interval = 0.82 to 3.42, p = 0.15). The types of complications that occurred were similar in both groups. While the data suggest an increased rate of post-extraction complications in the HIV-positive group, the difference in complication rates between the two groups was not statistically significant. In addition, the complications were minor, self-limiting, and readily treated. Based on these findings, we believe that tooth extraction is a low-risk procedure in HIV positive patients. Treatment may be rendered routinely to patients who present on an outpatient basis without the need for an extensive pre-operative work-up, unless otherwise indicated by relevant history. PMID- 9326898 TI - Changes in the shape and orientation of periodontal ligament fibroblasts in the continuously erupting rat incisor following removal of the occlusal load. AB - One of the main theories which attempts to explain the phenomenon of tooth eruption suggests that periodontal ligament (PDL) fibroblasts move actively and pull the tooth with them out of its socket. To find further support for this theory, we determined the changes in the shape and orientation of PDL fibroblasts induced by a transition from impeded to unimpeded eruption. We measured nuclear area, elongation (length-to-width ratio), and orientation (angulation in relation to the eruption axis) of PDL fibroblasts in impeded (functionally loaded) and unimpeded (hypoloaded) rat incisors. The mean cross-sectional nuclear area did not differ between fibroblasts in the two groups. In contrast, unimpeded eruption resulted in a marked increase in the mean nuclear elongation (from about 2 to 2.56) and a significant increase in the mean nuclear orientation (from 25.6 to 14.0 degrees). Bivariate analysis suggested that these changes occurred in the same cells. Analysis of nuclear elongation and orientation at various distances from the cementum toward the alveolar bone revealed a profile of both parameters, such that cells located 20 to 80 microns away from the cemental surface were more elongated and more frequently oriented toward the eruption axis, while cells at 0 to 20 and 80 to 100 microns were more round/oval and had a greater angulation with the eruption axis. These findings, together with other observations of changes in cell number, number of microtubules, and migration velocity which occur on the shift to unimpeded eruption, support the theory of active movement of PDL fibroblasts as an important component of tooth eruption. PMID- 9326897 TI - Smoking, smoking cessation, and tooth loss. AB - Smoking is associated with an increased risk of tooth loss, but it is not known if this risk decreases significantly when individuals quit smoking. The objectives of this study were to describe the rates of tooth loss by smoking status in two populations of medically healthy men and women. Among the men, rates of tooth loss and edentulism in relation to smoking cessation were also evaluated. The subjects were drawn from a group of 584 women (aged 40 to 70) recruited from the Boston, MA, area and a separate population of 1231 male veterans (aged 21 to 75) who participated in the VA Dental Longitudinal Study in Boston. In cross-sectional baseline analyses, current cigarette smokers of either sex had significantly more missing teeth than never-smokers or former smokers. Former smokers and pipe or cigar smokers tended to have an intermediate number of missing teeth. Current male smokers had more teeth with calculus, but the differences in plaque, tooth mobility, probing depth > 2 mm, filled and decayed teeth, and bleeding on probing by smoking history were not significant. Prospective observations of 248 women (mean follow-up time = 6 +/- 2 years) and 977 men (mean = 18 +/- 7 years) indicated that individuals who continued to smoke cigarettes had 2.4-fold (men) to 3.5-fold risk (women) of tooth loss compared with non-smokers. The rates of tooth loss in men were significantly reduced after they quit smoking cigarettes but remained higher than those in non-smokers. Men who smoked cigarettes had a 4.5-fold increase in risk of edentulism, and this risk also decreased upon smoking cessation. These findings indicate that the risk of tooth loss is greater among cigarette smokers than among non-smokers. Smoking cessation significantly benefits an individual's likelihood of tooth retention, but it may take decades for the individual to return to the rate of tooth loss observed in non-smokers. PMID- 9326899 TI - Association between marginal bone loss around osseointegrated mandibular implants and smoking habits: a 10-year follow-up study. AB - While many factors are conceivable, occlusal loading and plaque-induced inflammation are frequently stated as the most important ones negatively affecting the prognosis of oral implants. Currently, little is known about the relative importance of such factors. The aim of this study was to analyze the influence of smoking and other possibly relevant factors on bone loss around mandibular implants. The participants were 45 edentulous patients, 21 smokers and 24 non-smokers, who were followed for 10-year period after treatment with a fixed implant-supported prosthesis in the mandible. The peri-implant bone level was measured on intraoral radiographs, information about smoking habits was based on a careful interview, and oral hygiene was evaluated from clinical registration of plaque accumulation. Besides standard statistical methods, multiple linear regression models were constructed for estimation of the relative influence of some factors on peri-implant bone loss. The long-term results of the implant treatment were good, and only three implants (1%) were lost. The mean marginal bone loss around the mandibular implants was very small, about 1 mm for the entire 10-year period. It was greater in smokers than in non-smokers and correlated to the amount of cigarette consumption. Smokers with poor oral hygiene showed greater marginal bone loss around the mandibular implants than those with good oral hygiene. Oral hygiene did not significantly affect bone loss in non smokers. Multivariate analyses showed that smoking was the most important factor among those analyzed for association with peri-implant bone loss. The separate models for smokers and non-smokers revealed that oral hygiene had a greater impact on peri-implant bone loss among smokers than among non-smokers. This study showed that smoking was the most important factor affecting the rate of peri implant bone loss, and that oral hygiene also had an influence, especially in smokers, while other factors, e.g., those associated with occlusal loading, were of minor importance. These results indicate that smoking habits should be included in analyses of implant survival and peri-implant bone loss. PMID- 9326901 TI - Computer intuition: guiding scientific research in imaging and oral implantology. AB - As the costs associated with clinical research and new drug development increase, it is incumbent upon us to develop alternative research methodologies. A new Computer Intuition (CI) program identifies literature-based evidence with the potential to generate a hypothesis that is most likely to be clinically confirmed regarding the cause of a disease or the questions being posed. The objective of this study was to demonstrate whether computer intuition can be used to guide scientific research in solving important biomedical questions. The establishment of CI as an alternative research methodology has the potential to accelerate the translation of basic science findings into clinical practice. The specific aim of this study was to develop a CI-driven hypothesis related to the controversial issue debating which radiographic imaging technology is most suitable for diagnostic purposes prior to the placement of oral implants. This hypothesis was then compared, in retrospect, with a known opinion established following a recent literature review on this issue. In his review of the literature, Frederiksen (1995) suggests that multiplanar reformatted computed tomography (CT) is the diagnostic imaging modality of choice prior to implant placement. To compare the CI hypothesis with Frederiksen's (1995) opinion, we subjected 34 relevant papers to CI analysis. The output consisted of clusters of important statements, phrases, and thought processes from the given dataset rated with the greatest potential to generate testable hypotheses. Both CI's output and Frederiksen (1995) indicated that CT scanning is the diagnostic imaging modality of choice prior to implant placement. Although the ultimate utility of CI is dependent on the successful testing of its derived hypothesis, this preliminary retrospective study suggests that CI might be useful in guiding scientific research. PMID- 9326900 TI - A within-subject comparison of mandibular long-bar and hybrid implant-supported prostheses: psychometric evaluation and patient preference. AB - Although it has been shown that patients are more satisfied with prostheses supported by implants than with conventional dentures, there have been few direct comparisons of the various designs of implant-supported prostheses. This within subject crossover clinical trial was designed to compare two forms of removable prostheses which are frequently prescribed for the edentulous mandible: a long bar overdenture supported by 4 implants and a two-implant hybrid overdenture. Sixteen completely edentulous subjects were given a new maxillary conventional denture: Ten of them received the mandibular long-bar prosthesis first and six the hybrid. After a two-month adaptation period, psychometric measures of various aspects of the prostheses and physiological tests of masticatory efficiency were carried out over three weeks. The mandibular prostheses were then changed and the procedures repeated. At the end of the study, subjects were asked to choose the mandibular prosthesis that they wished to keep, and final psychometric measures were taken. In this paper, the results of the psychometric assessment and patient preference are presented. Subjects assessed factors such as general satisfaction, quality of life, stability, retention, comfort, esthetics, ease of cleaning, speaking, and chewing, and how well-chewed foods were before being swallowed. Most of the factors except ease of cleaning and speaking were rated significantly better with long-bar overdentures than with hybrid ones. These results are consistent with the fact that all subjects chose long-bar overdentures, reporting stability, ease of chewing, and comfort as the most important factors influencing their choice. These results suggest that, although subjects assign high ratings for most factors to hybrid overdentures, they find long-bar overdentures to be significantly more stable, comfortable, and easier for chewing. PMID- 9326902 TI - A three-dimensional finite element model of prismatic enamel: a re-appraisal of the data on the Young's modulus of enamel. AB - The inconsistencies of published data on the Young's modulus of dental enamel, the parameter used to quantify stiffness, have, for a long time, restricted our understanding of the biomechanical behavior of teeth. With the use of modeling techniques, the aim of this paper is to investigate which of the data may be more reliable. In this way, the possible causes of the discrepancies in data will be addressed. Two different structural levels are considered within the model. At an ultrastructural (i.e., crystalline) level, the model considers enamel to behave as a simple composite, being made up of long, parallel crystals held together by an organic matrix. At this level, the stiffness of enamel is predicted by simple composite theory, and the model indicates that stiffness is dependent on chemical composition and crystal orientation. At a microstructural (i.e., prismatic) level, the model considers enamel to behave as a hierarchical composite, being made up of prisms, in which the crystal orientation is heterogeneous. At this level, the stiffness of enamel is predicted by finite element stress analysis, and values of predicted stiffness are found to be dependent on both chemical composition and prism orientation. Within a realistic compositional range, predicted values of Young's modulus along the direction of prisms are comparable with the corresponding experimental values of 77.9 +/- 4.8 GPa obtained by Craig et al. (1961) and 73 GPa obtained by Gilmore et al. (1970), but not with those low values of 9.65 +/- 3.45 obtained by Stanford et al. (1960). Predictions of Young's modulus values across the direction of prisms are also made, and the model is less stiff in this direction. These findings indicate that human prismatic enamel is almost certainly anisotropic with respect to stiffness. PMID- 9326903 TI - Enamel subsurface damage due to tooth preparation with diamonds. AB - In clinical tooth preparation with diamond burs, sharp diamond particles indent and scratch the enamel, causing material removal. Such operations may produce subsurface damage in enamel. However, little information is available on the mechanisms and the extent of subsurface damage in enamel produced during clinical tooth preparation. The aim of this study, therefore, was to investigate the mechanisms of subsurface damage produced in enamel during tooth preparation by means of diamond burs, and to examine the dependence of such damage on enamel rod orientation, diamond particle size, and removal rate. Subsurface damage was evaluated by a bonded-interface technique. Tooth preparation was carried out on two enamel rod orientations, with four clinical diamond burs (coarse, medium, fine, and superfine) used in a dental handpiece. The results of this study showed that subsurface damage in enamel took the form of median-type cracks and distributed microcracks, extending preferentially along the boundaries between the enamel rods. Microcracks within individual enamel rods were also observed. The median-type cracks were significantly longer in the direction parallel to the enamel rods than perpendicular to the rods. Preparation with the coarse diamond bur produced cracks as deep as 84 +/- 30 microns in enamel. Finishing with fine diamond burs was effective in crack removal. The crack lengths in enamel were not significantly different when the removal rate was varied. Based on these results, it is concluded that subsurface damage in enamel induced by tooth preparation takes the form of median-type cracks as well as inter- and intra-rod microcracks, and that the lengths of these cracks are sensitive to diamond particle size and enamel rod orientation, but insensitive to removal rate. PMID- 9326904 TI - On the sensitivity of fecal occult blood test screening for colorectal cancer. PMID- 9326905 TI - The story of second cancers in patients cured of testicular cancer: tarnishing success or burnishing irrelevance? PMID- 9326906 TI - A tale of too witty? Using whimsy to name fringe genes. PMID- 9326907 TI - Greying of America will foster new strategies in oncology. PMID- 9326908 TI - Researchers test hi-tech bra for detecting breast cancer. PMID- 9326909 TI - Heat-seeking pads may help find early breast cancers. PMID- 9326910 TI - Participation in colorectal cancer screening: a review. AB - The purpose of this review is to evaluate the published literature on adherence to colorectal cancer (CRC) screening with fecal occult blood testing (FOBT) and sigmoidoscopy. Specifically, the review addresses the following: 1) prevalence of FOBT and sigmoidoscopy; 2) interventions to increase adherence to FOBT and sigmoidoscopy; 3) correlates or predictors of adherence to FOBT and sigmoidoscopy; and 4) reasons for nonadherence. Other objectives are to put the literature on CRC screening adherence in the context of recently reported findings from experimental interventions to change prevention and early detection behaviors and to suggest directions for future research on CRC screening adherence. CRC screening offers the potential both for primary and for secondary prevention. Data from the 1992 National Health Interview Survey show that 26% of the population more than 49 years of age report FOBT within the past 3 years and 33% report ever having had sigmoidoscopy. The Year 2000 goals set forth in Healthy People 2000 are for 50% of the population more than 49 years of age to report FOBT within the past 2 years and for 40% to report that they ever had sigmoidoscopy. Thus, systematic efforts to increase CRC screening are warranted. To date, attempts to promote CRC screening have used both a public health model that targets entire communities, e.g., mass media campaigns, and a medical model that targets individuals, e.g., general practice patients. Most of these efforts, however, did not include systematic evaluation of strategies to increase adherence. The data on FOBT show that the median adherence rate to programmatic offers of FOBT is between 40% and 50%, depending on the type of population offered the test, e.g., patients or employees. Approximately, 50% of those initially offered testing in unselected populations will respond to minimal prompts or interventions. A salient issue for FOBT, however, is whether or not the behavior can be sustained over time. Fewer studies examined adherence to sigmoidoscopy. Adherence was highest in relatives of CRC cases and in employer sponsored programs offered to workers at increased risk of CRC. At present, we know very little about the determinants of CRC screening behaviors, particularly as they relate to rescreening. PMID- 9326911 TI - Fecal occult blood screening in the Minnesota study: role of chance detection of lesions. AB - BACKGROUND: In the Minnesota Colon Cancer Control Study, annual fecal occult blood testing reduced mortality from colorectal cancer by at least 33.4%. Some attribute a large part of this reduction to chance detection of cancers by colonoscopies; rehydration of guaiac test slides greatly increased positivity and consequently the number of colonoscopies performed. This study was conducted to determine how much of the reduction resulted from chance detection. METHODS: We used a mathematical model developed by Lang and Ransohoff to estimate the proportion of the 33.4% mortality attainable by chance alone. Applying the model requires the specification of five parameters: duration of follow-up, rate of compliance with fecal occult blood testing, rate of compliance with colonoscopy, positivity rate, and efficacy of colonoscopy in reducing colorectal cancer mortality. We took values for four of the five parameters directly from the Minnesota study. For the fifth parameter, efficacy of colonoscopy, we selected a value of 60%, based on the conclusions of another study. Whereas the Lang Ransohoff model selects persons for colonoscopy by chance alone, those with bleeding cancers would also be selected by sensitive fecal occult blood testing. We therefore adjusted the result of the Lang-Ransohoff model for this dual detectability. RESULTS: We found that 16%-25% of the reduction in colorectal cancer deaths effected by fecal occult blood testing in the Minnesota study was due to chance detection; the remainder was due to sensitive detection. CONCLUSION: Chance played a minor role in the detection of colorectal cancers by fecal occult blood testing in the Minnesota study. PMID- 9326912 TI - Risk of second malignant neoplasms among long-term survivors of testicular cancer. AB - BACKGROUND: We have quantified the site-specific risk of second malignant neoplasms among nearly 29,000 survivors (> or = 1 year) of testicular cancer, taking into account the histologic type of initial cancer and the primary therapy used to treat it. METHODS: The study cohort consisted of 28,843 men identified within 16 population-based tumor registries in North America and Europe; over 3300 men had survived more than 20 years. New invasive cancers were identified through a search of registry files. RESULTS: Second cancers were reported in 1406 men (observed-to-expected ratio [O/E] = 1.43; 95% confidence interval = 1.36 1.51), with statistically significant excesses noted for acute lymphoblastic leukemia (O/E = 5.20), acute nonlymphocytic leukemia (O/E = 3.07), melanoma (O/E = 1.69), non-Hodgkin's lymphoma (O/E = 1.88), and cancers of the stomach (O/E = 1.95), colon (O/E = 1.27), rectum (O/E = 1.41), pancreas (O/E = 2.21), prostate (O/E = 1.26), kidney (O/E = 1.50), bladder (O/E = 2.02), thyroid (O/E = 2.92), and connective tissue (O/E = 3.16). Overall risk was similar after seminomas (O/E = 1.42) or nonseminomatous tumors (O/E = 1.50). Risk of solid tumors increased with time since the diagnosis of testicular cancer, yielding an O/E = 1.54 (O = 369) among 20-year survivors (two-sided P for trend = .00002). Secondary leukemia was associated with both radiotherapy and chemotherapy, whereas excess cancers of the stomach, bladder, and, possibly, pancreas were associated mainly with radiotherapy. CONCLUSIONS: Men with testicular cancer continue to be at significantly elevated risk of second malignant neoplasms for more than two decades following initial diagnosis. Patterns of excess second cancers suggest that many factors may be involved, although the precise roles of treatment, natural history, diagnostic surveillance, and other influences are yet to be clarified. PMID- 9326913 TI - Fecal occult blood screening in the Minnesota study: sensitivity of the screening test. AB - BACKGROUND: In the Minnesota Colon Cancer Control Study, which used guaiac slides to annually screen stool samples for blood, mortality from colorectal cancer was reduced by 33.4%. The reported sensitivity of this test for colorectal cancer was about 90%. However, results from another study estimated the sensitivity to be 25%-33%; other investigators have reported intermediate values. Given these contradictions, we examined screening sensitivity for colorectal cancer in the Minnesota study by several direct and indirect methods. METHODS: In this reanalysis of data from the Minnesota study, we distinguished between sensitivity for colorectal cancer of the screening test (composed of six slides) and of the screening program (a series of such tests). We estimated screen sensitivity by adjusting the crude estimate from the final tests in each screening phase for colorectal cancer incidence in 5 years of follow-up, by modeling guaiac slide results at each screen as a function of the presence of occult blood, and by incorporating sensitive detection into a modification of a mathematical model developed by Lang and Ransohoff. Program sensitivity was estimated from the fraction of screen-detected cancers among all cancers diagnosed in screened individuals. RESULTS: The crude estimate of program sensitivity was 89.4%, whereas the modified Lang-Ransohoff model estimates screen sensitivities at 94.1% 96.2%, consistent with the estimates from the other methods. Indirect measures, such as the association between the number of positive slides among the six slides in each set and the positive predictivity for colorectal cancer, are consistent with these estimates. CONCLUSIONS: The Minnesota study reduced mortality from colorectal cancer through use of a screening test with average screen and program sensitivities of about 90%. PMID- 9326914 TI - Concordance of genetic alterations in poorly differentiated colorectal neuroendocrine carcinomas and associated adenocarcinomas. AB - BACKGROUND: The histopathologic spectrum of colorectal neuroendocrine tumors ranges from benign to highly malignant. In this spectrum, poorly differentiated neuroendocrine carcinoma (PDNC) is the most aggressive type, characterized by early dissemination and a rapidly fatal course. Since it is unclear whether PDNC originates from neoplastic transformation of preexisting neuroectodermal cells, pluripotent epithelial stem cells, or adenocarcinoma precursor cells, we investigated the histogenesis of this type of cancer by performing genetic analyses on a series of colorectal tumors. METHODS: Archived histologic sections of colorectal PDNC from nine patients were analyzed; gastrointestinal carcinoid tumor specimens from four patients were used as controls. The specimens were deparaffinized, microdissected, and analyzed genetically. After DNA extraction, polymerase chain reaction amplification was performed to investigate alteration (i.e., loss of heterozygosity [LOH]) of the APC (adenomatous polyposis coli), DCC (deleted in colorectal carcinoma), and p53 (also known as TP53) genes. RESULTS: LOH of the APC, DCC, or p53 genes was observed in six of eight informative PDNC tumors; no LOH was detected in the carcinoid control specimens. Four of five informative PDNC tumors had associated adenocarcinoma; LOH of the APC and p53 genes in these tumors involved the same allele in both tissue components. Four of the five tumors with associated adenocarcinoma showed LOH of the DCC gene; in three of these four tumors, the PDNC and adenomatous components showed LOH of the same allele. CONCLUSIONS: PDNC and associated adenocarcinoma appear to be derived from the same cell of origin, which is most likely either a pluripotent epithelial stem cell or an adenocarcinoma precursor cell. PMID- 9326915 TI - Risk of urinary tract cancers following kidney or ureter stones. AB - BACKGROUND: A relationship has been suggested between kidney or ureter stones and the development of urinary tract cancers. In this study, a population-based cohort of patients hospitalized for kidney or ureter stones in Sweden was followed for up to 25 years to examine subsequent risks for developing renal cell, renal pelvis/ureter, or bladder cancer. METHODS: Data from the national Swedish In-patient Register and the national Swedish Cancer Registry were linked to follow 61,144 patients who were hospitalized for kidney or ureter stones from 1965 through 1983. Standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) were computed on the basis of nationwide cancer incidence rates, after adjustment for age, sex, and calendar year. RESULTS: Risk of renal cell cancer was not elevated in this cohort. Significant excesses of renal pelvis/ureter cancer (SIR = 2.5; 95% CI = 1.8-3.3) and bladder cancer (SIR = 1.4; 95% CI = 1.3-1.6) were observed, but the SIRs for women were more than twice those for men. Risks varied little by age or duration of follow-up. Risks of renal pelvis/ureter cancer and bladder cancer among patients with an associated diagnosis of urinary tract infection were more than double those among patients without such infection, although the risks were significantly elevated in both groups. CONCLUSIONS: Individuals hospitalized for kidney or ureter stones are at increased risk of developing renal pelvis/ureter or bladder cancer, even beyond 10 years of follow-up. Chronic irritation and infection may play a role, since kidney or ureter stones were located on the same side of the body as the tumors in most patients with renal pelvis/ureter cancer evaluated in our study. PMID- 9326916 TI - Can acid phosphatase reduce pap test false-negative readings? PMID- 9326917 TI - Germline CDKN2A mutations in childhood melanoma. PMID- 9326918 TI - Come together. PMID- 9326919 TI - Chloride channels cough up. PMID- 9326920 TI - The maddening hunt for madness genes. PMID- 9326921 TI - Molecular evolution--who is in the driver's seat? PMID- 9326922 TI - On the TRAIL from p53 to apoptosis? PMID- 9326923 TI - A searchlight through the fog. PMID- 9326924 TI - A human compound heterozygote for two MLH1 missense mutations. PMID- 9326925 TI - Sex-dependent rearrangements resulting in CMT1A and HNPP. PMID- 9326926 TI - Early-onset type-II diabetes mellitus (MODY4) linked to IPF1. PMID- 9326927 TI - Mutations in RPE65 cause Leber's congenital amaurosis. PMID- 9326928 TI - KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene. PMID- 9326929 TI - PTEN1 is frequently mutated in primary endometrial carcinomas. PMID- 9326930 TI - Genetic interaction between HAP1/REF-1 and p53. PMID- 9326931 TI - Mutations in transcriptional regulator ATRX establish the functional significance of a PHD-like domain. PMID- 9326932 TI - Diabetes, dependence, asymptotics, selection and significance. PMID- 9326933 TI - A novel gene encoding an SH3 domain protein is mutated in nephronophthisis type 1. AB - Juvenile nephronophthisis (NPH), an autosomal recessive cystic kidney disease, is the primary genetic cause of chronic renal failure in children. About two thirds of patients with NPH carry a large homozygous deletion at the gene locus NPH1 on 2q13. We here identify a novel gene. NPHP1, which extends over most of this common deletion. The 4.5-kb transcript encodes a protein with an SH3 domain, which is highly conserved throughout evolution. The 11-kb interval between the 3' end of NPHP1 and an inverted repeat containing the distal deletion breakpoint was found to contain the first exon of a second gene, MALL. In patients with a hemizygous deletion of the NPH1 region, additional point mutations were found in NPHP1 but not in MALL. PMID- 9326934 TI - Homologous recombination of a flanking repeat gene cluster is a mechanism for a common contiguous gene deletion syndrome. AB - Smith-Magenis syndrome (SMS), caused by del(17)p11.2, represents one of the most frequently observed human microdeletion syndromes. We have identified three copies of a low-copy-number repeat (SMS-REPs) located within and flanking the SMS common deletion region and show that SMS-REP represents a repeated gene cluster. We have isolated a corresponding cDNA clone that identifies a novel junction fragment from 29 unrelated SMS patients and a different-sized junction fragment from a patient with dup(17)p11.2. Our results suggest that homologous recombination of a flanking repeat gene cluster is a mechanism for this common microdeletion syndrome. PMID- 9326935 TI - Positional cloning of the gene associated with X-linked juvenile retinoschisis. AB - X-linked juvenile retinoschisis(RS) is a recessively inherited vitreo-retinal degeneration characterized by macular pathology and intraretinal splitting of the retina. The RS gene has been localized to Xp22.2 to an approximately 1 Mb interval between DXS418 and DXS999/DXS7161. Mapping and expression analysis of expressed sequence tags have identified a novel transcript, designated XLRS1, within the centromeric RS locus that is exclusively expressed in retina. The predicted XLRS1 protein contains a highly conserved motif implicated in cell-cell interaction and thus may be active in cell adhesion processes during retinal development. Mutational analyses of XLRS1 in affected individuals from nine unrelated RS families revealed one nonsense, one frameshift, one splice acceptor and six missense mutations segregating with the disease phenotype in the respective families. These data provide strong evidence that the XLRS1 gene, when mutated, causes RS. PMID- 9326936 TI - Mutations in the chloride channel gene, CLCNKB, cause Bartter's syndrome type III. AB - Analysis of patients with inherited hypokalaemic alkalosis resulting from salt wasting has proved fertile ground for identification of essential elements of renal salt homeostasis and blood-pressure regulation. We now demonstrate linkage of this phenotype to a segment of chromosome 1 containing the gene encoding a renal chloride channel, CLCNKB. Examination of this gene reveals loss-of-function mutations that impair renal chloride reabsorption in the thick ascending limb of Henle's loop. Mutations in seventeen kindreds have been identified, and they include large deletions and nonsense and missense mutations. Some of the deletions are shown to have arisen by unequal crossing over between CLCNKB and the nearby related gene, CLCNKA. Patients who harbour CLCNKB mutations are characterized by hypokalaemic alkalosis with salt-wasting, low blood pressure, normal magnesium and hyper- or normocalciuria; they define a distinct subset of patients with Bartter's syndrome in whom nephrocalcinosis is absent. These findings demonstrate the critical role of CLCNKB in renal salt reabsorption and blood-pressure homeostasis, and demonstrate the potential role of specific CLCNKB antagonists as diuretic antihypertensive agents. PMID- 9326937 TI - Perinatal lethality with kidney and pancreas defects in mice with a targetted Pkd1 mutation. AB - PKD1 is the most common site for mutations in human autosomal dominant polycystic kidney disease (ADPKD). ADPKD is characterized by progressive replacement of kidney tissue by epithelial cysts and eventual renal failure. Hepatic and pancreatic cysts are also common. The PKD1 protein, polycystin, is a cell-surface protein of unknown function that is widely expressed in epithelia and in vascular smooth muscle and myocardium. None of the genetic forms of murine polycystic disease map to the murine Pkd1 locus. We introduced into mice by homologous recombination a Pkd1 truncation mutation, Pkd1-, that mimics a mutation found in ADPKD. Pkd1- heterozygotes have no discernible phenotype, whereas homozygotes die during the perinatal period with massively enlarged cystic kidneys, pancreatic ductal cysts and pulmonary hypoplasia. Renal cyst formation begins at embryonic day 15.5 (E15.5) in proximal tubules and progresses rapidly to replace the entire renal parenchyma. The timing of cyst formation indicates that full-length polycystin is required for normal morphogenesis during elongation and maturation of tubular structures in the kidney and pancreas. PMID- 9326938 TI - Male-driven evolution of DNA sequences in birds. AB - Assuming that new mutations arise mainly during DNA replication, sequence evolution in mammals has been seen as 'male driven' (ref. 1) because of the many more cell divisions in spermatogenesis than in oogenesis. Molecular support for this idea has been obtained from the observation of higher substitution rates in genes on the Y than on the X chromosome of primates and rodents, which are species with male heterogamety, but has not been confirmed by the reciprocal analysis of organisms with female heterogamety. The recent suggestion that an intrinsic reduction in the X-chromosome mutation rate may be confounded with male effects in previous comparisons, and the paradoxical finding of low levels of polymorphism on the primate Y chromosome indicate that the idea of male-biased mutation rate needs to be re-examined. We have analysed the molecular evolution of the gene CHD, which is present on the Z and W sex chromosomes of birds. The substitution rate at synonymous positions, as well as in intron DNA, was considerably higher on the Z chromosome than on the female-specific W chromosome, with an estimated male-to-female bias in mutation rate (alpha m) of 3.9-6.5. Thus, evolution appears to be male driven in birds--a situation that supports a neutral model of molecular evolution. PMID- 9326939 TI - Identification of PAHX, a Refsum disease gene. AB - Refsum disease is an autosomal recessive disorder characterized by retinitis pigmentosa, peripheral polyneuropathy, cerebellar ataxia and increased cerebrospinal fluid protein. Biochemically, the disorder is defined by two related properties: pronounced accumulation of phytanic acid and selective loss of the peroxisomal dioxygenase required for alpha-hydroxylation of phytanoyl CoA2. Decreased phytanic-acid oxidation is also observed in human cells lacking PEX7, the receptor for the type-2 peroxisomal targetting signal (PTS2; refs 3,4), suggesting that the enzyme defective in Refsum disease is targetted to peroxisomes by a PTS2. We initially identified the human PAHX and mouse Pahx genes as expressed sequence tags (ESTs) capable of encoding PTS2 proteins. Human PAHX is targetted to peroxisomes, requires the PTS2 receptor for peroxisomal localization, interacts with the PTS2 receptor in the yeast two-hybrid assay and has intrinsic phytanoyl-CoA alpha-hydroxylase activity that requires the dioxygenase cofactor iron and cosubstrate 2-oxoglutarate. Radiation hybrid data place PAHX on chromosome 10 between the markers D10S249 and D10S466, a region previously implicated in Refsum disease by homozygosity mapping. We find that both Refsum disease patients examined are homozygous for inactivating mutations in PAHX, demonstrating that mutations in PAHX can cause Refsum disease. PMID- 9326940 TI - Refsum disease is caused by mutations in the phytanoyl-CoA hydroxylase gene. AB - Refsum disease is an autosomal-recessively inherited disorder characterized clinically by a tetrad of abnormalities: retinitis pigmentosa, peripheral neuropathy, cerebellar ataxia and elevated protein levels in the cerebrospinal fluid (CSF) without an increase in the number of cells in the CSF. All patients exhibit accumulation of an unusual branched-chain fatty acid, phytanic acid (3,7,11,15-tetramethylhexadecanoic acid), in blood and tissues. Biochemically, the disease is caused by the deficiency of phytanoyl-CoA hydroxylase (PhyH), a peroxisomal protein catalyzing the first step in the alpha-oxidation of phytanic acid. We have purified PhyH from rat-liver peroxisomes and determined the N terminal amino-acid sequence, as well as an additional internal amino-acid sequence obtained after Lys-C digestion of the purified protein. A search of the EST database with these partial amino-acid sequences led to the identification of the full-length human cDNA sequence encoding PhyH: the open reading frame encodes a 41.2-kD protein of 338 amino acids, which contains a cleavable peroxisomal targeting signal type 2 (PTS2). Sequence analysis of PHYH fibroblast cDNA from five patients with Refsum disease revealed distinct mutations, including a one nucleotide deletion, a 111-nucleotide deletion and a point mutation. This analysis confirms our finding that Refsum disease is caused by a deficiency of PhyH. PMID- 9326941 TI - Mutations in RPE65 cause autosomal recessive childhood-onset severe retinal dystrophy. AB - Autosomal recessive childhood-onset severe retinal dystrophy (arCSRD) designates a heterogeneous group of disorders affecting rod and cone photoreceptors simultaneously. The most severe cases are termed Leber congenital amaurosis (LCA), while the less aggressive forms are usually considered juvenile retinitis pigmentosa. Recently, mutations in the retinal-specific guanylate cyclase gene were found in patients with LCA. Disease genes implicated in other forms of arCSRD are expected to encode proteins present in the neuroretina or in the retinal pigment epithelium (RPE). The RPE, a monolayer of cells separating the vascular-rich choroid and the neuroretina, is in intimate contact with the outer segments of rods and cones via the microvilli surrounding the photoreceptors. The RPE expresses a tissue-specific and evolutionarily highly conserved 61 kD protein (RPE65) present at high levels in vivo. Although the function of RPE65 is not yet known, an important role in the RPE/photoreceptor vitamin-A cycle is suggested by the fact that RPE65 associates both with serum retinol-binding protein and with the RPE-specific 11-cis retinol dehydrogenase, an enzyme active in the synthesis of the visual pigment chromophore 11-cis retinal. Here we report that the analysis of RPE65 in a collection of about 100 unselected retinal-dystrophy patients of different ethnic origin revealed five that are likely to be pathogenic mutations, including a missense mutation (Pro363Thr), two point mutations affecting splicing (912 + 1G-->T and 65 + 5G-->A) and two small re arrangements (ins144T and 831del8) on a total of nine alleles of five patients with arCSRD. In contrast to other genes whose defects have been implicated in degenerative retinopathies, RPE65 is the first disease gene in this group of inherited disorders that is expressed exclusively in the RPE, and may play a role in vitamin-A metabolism of the retina. PMID- 9326942 TI - Mutation of the gene encoding cellular retinaldehyde-binding protein in autosomal recessive retinitis pigmentosa. AB - Inadequate levels of all-trans-retinol in the blood cause retinal dysfunction; hence, genes implicated in retinal vitamin-A metabolism represent candidates for inherited retinal degenerations. In the current study, molecular genetic analysis of a consanguineous pedigree segregating for non-syndromic autosomal recessive retinitis pigmentosa (arRP) indicated that the affected siblings were homozygous by descent for a G4763A nucleotide substitution in RLBP1, the gene encoding cellular retinaldehyde-binding protein (CRALBP). This substitution is predicted to replace an arginine with glutamine at residue 150. CRALBP is not expressed in photoreceptors but is abundant in the retinal pigment epithelium (RPE) and Muller cells of the neuroretina, where it carries 11-cis-retinol and 11-cis retinaldehyde. When expressed in bacteria, recombinant CRALBP (rCRALBP) containing the R150Q substitution was less soluble than wild-type rCRALBP. Mutant rCRALBP was purified from the soluble cell lysate and the protein structure was verified by mass spectrometry. The mutant protein lacked the ability to bind 11 cis-retinaldehyde. These findings suggest that arRP in the current pedigree results from a lack of functional CRALBP, presumably leading to disruption of retinal vitamin-A metabolism. PMID- 9326943 TI - The genetic and functional basis of isolated 17,20-lyase deficiency. AB - Human male sexual differentiation requires production of fetal testicular testosterone, whose biosynthesis requires steroid 17,20-lyase activity. Patients with putative isolated 17,20-lyase deficiency have been reported. The existence of true isolated 17,20-lyase deficiency, however, has been questioned because 17 alpha-hydroxylase and 17,20-lyase activities are catalyzed by a single enzyme, microsomal cytochrome P450c17, and because the index case of apparent isolated 17,20-lyase deficiency had combined deficiencies of both activities. We studied two patients with clinical and hormonal findings suggestive of isolated 17,20 lyase deficiency. We found two patients homozygous for substitution mutations in CYP17, the gene encoding P450c17. When expressed in COS-1 cells, the mutants retained 17 alpha-hydroxylase activity but had minimal 17,20-lyase activity. Substrate competition experiments suggested that the mutations did not alter the enzyme's substrate-binding capacity, but co-transfection of cells with P450 oxidoreductase, the electron donor used by P450c17, indicated that the mutants had a diminished ability to interact with redox partners. Computer-graphic modelling of P450c17 suggests that both mutations lie in or near the redox partner binding site, on the opposite side of the haem from the substrate-binding pocket. These mutations alter electrostatic charge distribution in the redox partner binding site, so that electron transfer for the 17,20-lyase reaction is selectively lost or diverted to uncoupling reactions. These are the first proven cases of isolated 17,20-lyase deficiency, and they demonstrate a novel mechanism for loss of enzymatic activity. PMID- 9326944 TI - Increased stress response and beta-phenylethylamine in MAOB-deficient mice. AB - MAOA and MAOB are key iso-enzymes that degrade biogenic and dietary amines. MAOA preferentially oxidizes serotonin (5-hydroxytryptamine, or 5-HT) and norepinephrine (NE), whereas MAOB preferentially oxidizes beta-phenylethylamine (PEA). Both forms can oxidize dopamine (DA). A mutation in MAOA results in a clinical phenotype characterized by borderline mental retardation and impaired impulse control. X-chromosomal deletions which include MAOB were found in patients suffering from atypical Norrie's disease, which is characterized by blindness and impaired hearing. Reduced MAOB activity has been found in type-II alcoholism and in cigarette smokers. Because most alcoholics smoke, the effects of alcohol on MAOB activity remain to be determined. Here we show that targetted inactivation of MAOB in mice increases levels of PEA but not those of 5-HT, NE and DA, demonstrating a primary role for MAOB in the metabolism of PEA. PEA has been implicated in modulating mood and affect. Indeed, MAOB-deficient mice showed an increased reactivity to stress. In addition, mutant mice were resistant to the neurodegenerative effects of MPTP, a toxin that induces a condition reminiscent of Parkinson's disease. PMID- 9326945 TI - Drosophila CBP is required for dorsal-dependent twist gene expression. AB - Although CREB-binding protein (CBP) functions as a co-activator of many transcription factors, relatively little is known about the physiological role of CBP. Mutations in the human CBP gene are associated with Rubinstein-Taybi syndrome, a haplo-insufficiency disorder characterized by abnormal pattern formation. Recently, we isolated a Drosophila CBP (dCBP) mutant, and found dCBP to be maternally expressed, suggesting that it plays a role in early embryogenesis. Mesoderm formation is one of the most important events during early embryogenesis. To initiate the differentiation of the mesoderm in Drosophila, multiple zygotic genes such as twist (twi) and snail (sna), which encode a basic-helix-loop-helix and a zinc finger transcription factor, respectively, are required. The transcription of these genes is induced by maternal dorsal (dl) protein (Dl; refs 8-10), a transcription factor that is homologous to the NF-kappa B family of proteins. The activity of dl is negatively regulated by cactus (cact), a Drosophila homologue of I kappa B. Here, we show that dCBP mutants fail to express twi and generate twisted embryos. This is explained by results showing that dCBP is necessary for dl-mediated activation of the twi promoter. PMID- 9326946 TI - Aconitase and mitochondrial iron-sulphur protein deficiency in Friedreich ataxia. AB - Friedreich ataxia (FRDA) is a common autosomal recessive degenerative disease (1/50,000 live births) characterized by a progressive-gait and limb ataxia with lack of tendon reflexes in the legs, dysarthria and pyramidal weakness of the inferior limbs. Hypertrophic cardiomyopathy is observed in most FRDA patients. The gene associated with the disease has been mapped to chromosome 9q13 (ref. 3) and encodes a 210-amino-acid protein, frataxin. FRDA is caused primarily by a GAA repeat expansion within the first intron of the frataxin gene, which accounts for 98% of mutant alleles. The function of the protein is unknown, but an increased iron content has been reported in hearts of FRDA patients and in mitochondria of yeast strains carrying a deleted frataxin gene counterpart (YFH1), suggesting that frataxin plays a major role in regulating mitochondrial iron transport. Here, we report a deficient activity of the iron-sulphur (Fe-S) cluster containing subunits of mitochondrial respiratory complexes I, II and III in the endomyocardial biopsy of two unrelated FRDA patients. Aconitase, an iron-sulphur protein involved in iron homeostasis, was found to be deficient as well. Moreover, disruption of the YFH1 gene resulted in multiple Fe-S-dependent enzyme deficiencies in yeast. The deficiency of Fe-S-dependent enzyme activities in both FRDA patients and yeast should be related to mitochondrial iron accumulation, especially as Fe-S proteins are remarkably sensitive to free radicals. Mutated frataxin triggers aconitase and mitochondrial Fe-S respiratory enzyme deficiency in FRDA, which should therefore be regarded as a mitochondrial disorder. PMID- 9326947 TI - Targetting of the gene encoding fibrillin-1 recapitulates the vascular aspect of Marfan syndrome. AB - Aortic aneurysm and dissection account for about 2% of all deaths in industrialized countries; they are also components of several genetic diseases, including Marfan syndrome (MFS). The vascular phenotype of MFS results from mutations in fibrillin-1 (FBN1), the major constituent of extracellular microfibrils. Microfibrils, either associated with or devoid of elastin, give rise to a variety of extracellular networks in elastic and non-elastic tissues. It is believed that microfibrils regulate elastic fibre formation by guiding tropo-elastin deposition during embryogenesis and early post-natal life. Hence, vascular disease in MFS is thought to result when FBN1 mutations preclude elastic fibre maturation by disrupting microfibrillar assembly. Here we report a gene targetting experiment in mice that indicates that fibrillin-1 microfibrils are predominantly engaged in tissue homeostasis rather than elastic matrix assembly. This finding, in turn, suggests that aortic dilation is due primarily to the failure by the microfibrillar array of the adventitia to sustain physiological haemodynamic stress, and that disruption of the elastic network of the media is a secondary event. PMID- 9326948 TI - Cre-mediated chromosome loss in mice. AB - Chromosome loss in early human embryos is thought to cause a large proportion of spontaneous abortions; when it occurs in specific cell lineages in older embryos or adults, it can result in neoplasia. Although early embryonic chromosome loss can be modelled by breeding mice carrying robertsonian translocation chromosomes, there is currently no method for producing mice with tissue-specific monosomies. Here we demonstrate that DNA recombination mediated by the site-specific recombinase Cre causes loss of a chromosome carrying loxP sites (Cre recognition sites) in an inverted orientation. Thus, when male mice carrying a Y-linked transgene containing inverted loxP sites are mated with females carrying a cre gene that is obiquitously expressed in the early embryo, almost all their XY progeny lose the Y chromosome early in embryogenesis and develop as XO females. Because inverted loxP sites can be targetted to any mouse chromosome and mice can be produced that express cre in specific cell lineages, these data suggest a method for engineering tissue-specific loss of particular chromosomes to provide mouse models for human diseases caused by or associated with specific monosomies. PMID- 9326949 TI - Deregulation of MUM1/IRF4 by chromosomal translocation in multiple myeloma. AB - The pathogenesis of multiple myeloma (MM), an incurable tumour causing the deregulated proliferation of terminally differentiated B cells, is unknown. Chromosomal translocations (14q1) affecting band 14q32 and unidentified partner chromosomes are common in this tumour, suggesting that they may cause the activation of novel oncogenes. By cloning the chromosomal breakpoints in an MM cell line, we show that the 14q+ translocation represents a t(6;14)x(p25;q32) and that this aberration is recurrent in MM, as it was found in two of eleven MM cell lines. The translocation juxtaposes the immunoglobulin heavy-chain (IgH) locus to MUM1 (multiple myeloma oncogene 1)/IRF4 gene, a member of the interferon regulatory factor (IRF) family known to be active in the control of B-cell proliferation and differentiation. As a result, the MUM1/IRF4 gene is overexpressed--an event that may contribute to tumorigenesis, a MUM1/IRF4 has oncogenic activity in vitro. These findings identify a novel genetic alteration associated with MM, with implications for the pathogenesis and diagnostics of this tumour. PMID- 9326950 TI - Human telomeres contain two distinct Myb-related proteins, TRF1 and TRF2. AB - Human telomeres are composed of long arrays of TTAGGG repeats that form a nucleoprotein complex required for the protection and replication of chromosome ends. One component of human telomeres is the TTAGGG repeat binding factor 1 (TRF1), a ubiquitously expressed protein, related to the protooncogene Myb, that is present at telomeres throughout the cell cycle. Recent evidence has implicated TRF1 in the control of telomere length. TRF1 is proposed to be an inhibitor of telomerase, acting in cis to limit the elongation of individual chromosome ends. Here we report the cloning of TRF2, a distant homologue of TRF1 that carries a very similar Myb-related DNA-binding motif. Like TRF1, TRF2 was ubiquitously expressed, bound specifically to duplex TTAGGG repeats in vitro and localized to all human telomeres in metaphase chromosomes. TRF2 was shown to have an architecture similar to that of TRF1 in that it carries a C-terminal Myb motif and a large TRF1-related dimerization domain near its N terminus. However, the dimerization domains of TRF1 and TRF2 did not interact, suggesting that these proteins exist predominantly as homodimers. While having similar telomere binding activity and domain organization, TRF2 differed from TRF1 in that its N terminus was basic rather than acidic, and TRF2 was much more conserved than TRF1. The results indicate that the TTAGGG repeat arrays at the ends of human and mouse chromosomes bind to two related proteins. Because TRF1 and TRF2 showed significant differences, we suggest that these factors have distinct functions at telomeres. PMID- 9326951 TI - Telomeric localization of TRF2, a novel human telobox protein. AB - Natural chromosomal ends are stabilized by proteins that bind duplex telomeric DNA repeats. In human cells, the TTAGGG Repeat Factor 1 (TRF1) was identified by two independent studies, one screening for factors that bind duplex telomeric DNA and the other screening for proteins containing a particular Myb motif called the telobox, which is required for telomeric repeat recognition (Fig. 1a; refs 3-5). A second human open reading frame, orf2, contains a telobox sequence and encodes a polypeptide that specifically recognizes mammalian telomeric repeat DNA in vitro. We show that two proteins of 65 and 69 kD, expressed in HeLa cells, contain the orf2 telobox sequence. These proteins are collectively termed TRF2. Affinity-purified antibodies specific for anti-TRF2 label the telomeres of intact human chromosomes, strengthening the correlation between occurrence of telobox and telomere-repeat recognition in vivo. PMID- 9326952 TI - Mutations in the plakophilin 1 gene result in ectodermal dysplasia/skin fragility syndrome. AB - Members of the armadillo protein gene family, which includes plakoglobin and beta catenin, have important functions in cytoskeleton/cell membrane interactions. These proteins may act as linker molecules at adherens junctions and desmosomes at the plasma membrane; in addition, they may have pivotal roles in signal transduction pathways and significant effects on cell behaviour during development. Here, we describe the first human mutations in one of these dual function proteins, plakophilin 1 (band-6 protein; refs 8-10). The affected individual has a complete absence of immunostaining for plakophilin 1 in the skin and is a compound heterozygote for autosomal-recessively inherited premature termination codons of translation on both alleles of the plakophilin 1 gene (PKP1). Clinically, there are features of both cutaneous fragility and congenital ectodermal dysplasia affecting skin, hair and nails. There is no evidence of significant abnormalities in other epithelia or tissues. Desmosomes in the skin are small and poorly formed with widening of keratinocyte intercellular spaces and perturbed desmosome/keratin intermediate filament interactions. The molecular findings and clinical observations in this patient attest to the dual importance of plakophilin 1 in both cutaneous cell-call adhesion and epidermal morphogenesis. PMID- 9326953 TI - Imaging in pediatric urology. AB - A multitude of disorders of the genitourinary tract can occur in children. Although some entities may be diagnosed clinically, radiologic imaging is often necessary for diagnosis and management. The radiologic work-up has been discussed using a problem-oriented approach in five clinical situations: urinary tract infection, hydronephrosis or hydroureter, trauma, swollen scrotum, and hematuria. This discussion provides some general guidelines, although the evaluation of each child may need to be individualized depending on their specific clinical symptomatology. PMID- 9326954 TI - Common office problems in pediatric urology and gynecology. AB - The number of genital problems that pediatricians encounter is substantial. The most common ones have been reviewed in this article. Perhaps the most important point to reinforce is the appropriateness of nonintervention in uncircumcised boys whose foreskins have not become retractile during early school years. Without infections or pathologic phimosis, these boys do well, and most foreskins become retractile as they approach puberty. Abnormalities beyond those discussed or those not fitting the anticipated pattern probably warrant specialty referral. PMID- 9326955 TI - Enuresis and common voiding abnormalities. AB - Voiding problems, and in particular nocturnal enuresis, can usually be evaluated and managed without resorting to complex procedures or invasive tests. A good history with attention to toilet habits and the possible presence of infection can help distinguish patients who may have significant organic pathologic conditions who require further investigation. Wetting alarms are effective with a low recidivism rate but are noisy. DDAVP is effective, works rapidly, and is discrete but has a higher recidivism rate. Treatment is aimed at correcting any poor toilet habits and using the appropriate alarm device or medication. PMID- 9326956 TI - Urinary tract infections in children. Epidemiology, evaluation, and management. AB - Accurate documentation of UTIs in children is essential for proper evaluation and management. Urine cultures with multiple organisms or colony counts less than 50,000 to 100,000 CFU/ml should be considered suspect and require confirmation, particularly with clean-catch specimens. Children with well-documented UTIs should be evaluated based on their age and presenting symptoms. Infants and young children require imaging, usually with a cystogram and sonogram of the kidneys and bladder. Older girls with febrile UTIs and boys at any age should also be considered for urinary tract imaging. Renal cortical scintigraphy with 99mTc-DMSA has emerged as the imaging study of choice for acute pyelonephritis and renal scarring in children with UTIs. Treatment of UTIs in children ideally commences with culture-specific antimicrobial therapy, although treatment may be started in sick children before culture results are available. Short-course treatment (3-5 days) is sufficient for children with acute uncomplicated lower UTIs. Children with acute pyelonephritis require 10 to 14 days of antibiotics, which can be administered on an outpatient basis in older infants and children who are not toxic, as long as good compliance is expected. Patients with first-time UTIs who require imaging should be maintained on low-dose antibiotic prophylaxis until their workup is completed. Treatment of ABU does not seem necessary if the urinary tract is otherwise normal. Long-term antibiotic prophylaxis is indicated for children with frequent symptomatic recurrences of UTI and for those with known VUR. Diagnosis and treatment of underlying voiding dysfunction and constipation is an essential component of the successful management of UTIs in children. PMID- 9326957 TI - Vesicoureteral reflux. AB - The management options outlined earlier are based on the available treatment modalities; however, when a simple, successful, durable, minimally invasive method becomes available to treat vesicoureteral reflux, the approach likely will change. Endoscopic outpatient treatment of reflux has been available for about a decade. Treatment entails injection of a material into the submucosa at the refluxing ureteral orifice to bolster it, thus curing the problem. A suspension of microscopic size polytetrafluoroethylene (Teflon) particles has been used; however, its safety has been seriously questioned, as some evidence shows migration of the particles to other organ systems, including the central nervous system. More recently, cross-linked bovine collagen has been similarly used; however, it does not appear to be as durable. The use of other materials that are safe and will lead to long-term success are being studied. Chondrocytes and other nonbiologic materials, such as microspheres of bioglass and detachable balloons, are being evaluated. It is fairly certain that when a safe material is found, patients with mild to moderate reflux will be endoscopically treated upon recognition, thereby avoiding the use of long-term prophylaxis and periodic radiographic reevaluation. PMID- 9326958 TI - Hematuria. An integrated medical and surgical approach. AB - The need to perform a detailed work-up of microscopic hematuria is based on the following set of questions: Does the history or physical examination findings suggest systemic or renal disease? Is the patient able to acidify and concentrate urine? Is proteinuria present? Do other family members have hematuria or other renal problems? Does the microscopic analysis show casts, crystals, or WBCs? Are the RBCs eumorphic or dysmorphic? Using this scheme of questions, most children do not require laboratory tests or radiographic studies. In the case of gross or macroscopic hematuria, the initial evaluation may require only a urine culture, urine calcium-to-creatinine ratio, and renal and bladder sonography or a very detailed evaluation for renal parenchymal disease, stones, tumors, or anatomic abnormalities. In these instances, consultation with a pediatric nephrologist, urologist, or both is necessary. PMID- 9326959 TI - Cryptorchidism. Current concepts. AB - Cryptorchidism is the most common genitourinary disorder of childhood. Even though its incidence has changed only slightly over the years, the number of operations for cryptorchid testes has tripled. Better understanding of the natural history of cryptorchidism, as well as changes that occur in testicular histology both in the cryptorchid and the contralateral descended testis very early in life, are the cause. This experience has led us to advocate early orchiopexy as the optimum means of treatment. PMID- 9326960 TI - Benign and malignant pediatric scrotal masses. AB - Pediatric patients presenting with painless scrotal masses can be perplexing because of the long differential diagnosis. A careful plan based on the physical examination and sonogram findings localizes the mass to the testis or an extratesticular location. Sonography distinguishes solid from cystic lesions. Subsequent management is based on the location and nature of the mass. Intratesticular masses are assumed to be malignant, but testis-sparing surgery is possible in pediatric patients. Extratesticular cystic lesions are likely benign and are managed according to the specific diagnosis. Solid extratesticular lesions require exploration to establish the correct diagnosis. PMID- 9326961 TI - The acute scrotum. AB - Every boy with acute onset scrotal pain and swelling requires an immediate evaluation. Our protocol (Fig. 6) for the evaluation of these children is based on the history and physical examination combined with the selective use of imaging studies. When used appropriately, this protocol facilitates the rapid identification of children with torsion and minimizes the number of unnecessary scrotal explorations. When the duration of the pain is brief, and history and physical examination suggest that torsion is the most likely diagnosis, urgent surgical exploration without additional imaging studies is recommended. When it is not possible to definitely diagnose or exclude the diagnosis of testicular torsion, or when the duration of pain is greater than 12 hours, then diagnostic imaging can provide significant information. Color Doppler sonography is, in the authors' opinion, preferable to nuclear imaging for the evaluation of children with acute scrotums. When normal or increased blood flow is present, scrotal exploration is not required. When the study demonstrates decreased blood flow or does not provide a definite diagnosis, scrotal exploration is recommended. The authors recommend this approach because less than one third of these children have testicular torsion, and if routine scrotal exploration is performed for all boys with acute scrotums, a significant number of unnecessary surgical procedures will result. PMID- 9326962 TI - Abnormalities of the external genitalia. AB - This article discusses the general nuances of hypospadias, exstrophy/epispadias, and ambiguous genitalia. Embryologic considerations, etiologic factors, anatomy, associated anomalies, and timing of referrals and surgery are discussed. PMID- 9326963 TI - Antenatal hydronephrosis. Fetal and neonatal management. AB - As many as 1% of newborn infants have a prenatal diagnosis of hydronephrosis or significant renal pelvic dilation. Hydronephrosis often is caused by nonobstructive conditions. The likelihood of significant urologic pathology is directly related to the size of the fetal renal pelvis, and 90% with an anteroposterior diameter more than 2 cm need surgery or long-term urologic medical care. Following delivery, antibiotic prophylaxis should be administered and a renal sonogram and voiding cystourethrogram should be obtained. If there is grade 3 or 4 hydronephrosis, usually a diuretic renogram is recommended also. Pediatric urologic or pediatric nephrologic consultation usually is helpful in planing evaluation and treatment. Prenatal recognition of hydronephrosis allows neonatal diagnosis and treatment of urologic pathology, preventing complications of pyelonephritis and obstruction. PMID- 9326964 TI - Renal duplication and fusion anomalies. AB - Renal duplications and fusion anomalies in children often present challenging problems. The diagnostic evolution of these entities often consists of upper tract imaging to evaluate function and help diagnose obstruction, and lower tract imaging to assess reflux and at times voiding dynamics. The clinician needs to be aware of the variable presentations of these lesions, their evolution, and the therapeutic interventions that may be required to resolve problems resulting from them. PMID- 9326965 TI - Ophthalmic instrumentation. AB - The instruments utilized in ophthalmic surgery are described along with their intended uses. The sterilization and storage of surgical instruments are also discussed. To guide the surgeon in the selection of instruments, examples of instrument packs are included. PMID- 9326966 TI - Principles of microsurgery. AB - The surgical principles and techniques used in ophthalmic microsurgery differ considerably from those used in general surgery. Successful ophthalmic microsurgery requires that the surgeon understand not only the design and complexities of the operating microscope, but how tissues are affected by minute manipulations with microsurgical instruments. Furthermore, ophthalmic microsurgery requires a detailed understanding of how microsurgical techniques need to be adjusted to accommodate the unique features of ocular tissues such as conjunctiva, cornea, and lens. A diligent effort to master the principles of ophthalmic microsurgery is probably the single most important prerequisite to becoming an accomplished ophthalmic surgeon. PMID- 9326967 TI - Surgery of the adnexa. AB - Review of the functional anatomy of ocular adnexal tissues is followed by presentation of surgical procedures aimed at correcting conditions of the eyelids, conjunctiva, and third eyelids of small animals. Procedures used effectively by the primary author are described in detail including instances where combination procedures may be indicated. Some newer, recently described techniques are also briefly discussed. Illustrations of applied anatomy and multiple surgical techniques are provided by illustrator and coauthor Dr. Gheorghe Constantinescu. References are given to encourage readers to further explore alternative techniques focusing on the surgical correction of adnexal diseases of dogs and cats. PMID- 9326968 TI - Surgery of the cornea. AB - Corneal surgery is a common and essential part of veterinary ophthalmology and ranges from simple linear keratotomy for indolent ulcers to penetrating keratoplasty for restoration of optical clarity. Success in corneal surgery relies on an understanding of corneal anatomy, physiology and wound healing, meticulous attention to detail, microsurgical equipment and techniques, and use of appropriate preoperative, intraoperative, and postoperative medications. Surgical management of corneal disease is indicated for corneal ulceration, excision of a mass lesion, reconstructive procedures, therapeutic indications, optical restoration, and cosmetic purposes. PMID- 9326970 TI - Phacoemulsification. Technology and fundamentals. AB - The number one rule of phacoemulsification and aspiration cataract surgery is to know your machine. This chapter is designed to help the surgeon who is currently using phacoemulsification, or those who wish to understand more about technique, learn the basics and technology of the various types of phacoemulsification machines. Fluidics, pump design, handpiece mechanics, phacoemulsification needles, and fundamentals of phacoemulsification of cataracts will be reviewed. PMID- 9326969 TI - Surgery for glaucoma. AB - Most cases of glaucoma in small animals ultimately require surgical treatment for long-term control of intraocular pressure. Surgical procedures that have the potential to preserve vision in acute cases are categorized into those that reduce aqueous production (cyclodestructive techniques). Salvage procedures for irreversibly blind eyes include enucleation, implantation of an intraocular prosthesis, and pharmacologic ciliary body ablation. The indications, surgical technique, and complications of these procedures are discussed in this article. PMID- 9326972 TI - Surgery for lens instability. AB - Lens luxation is a common and potentially blinding disease of dogs. If left untreated, degenerative changes in the pathways for aqueous humor result in glaucoma; however, if the lens is removed by ICLE before significant secondary changes occur, vision can be preserved. In addition, it is now possible to restore excellent vision by replacing the luxated lens with a synthetic IOL. PMID- 9326971 TI - Surgery for cataracts. AB - The advent of phacoemulsification has substantially improved the success rate of cataract surgery in dogs, whereas the development of artificial lens implantation has equally improved postoperative visual acuity. In this chapter information pertaining to the etiology, diagnosis, and management of canine cataracts is provided for the general practitioner. More detailed information on phacoemulsification and artificial lens implantation is provided for residents in training or practicing ophthalmologists that may be converting from extracapsular extraction. PMID- 9326973 TI - Surgery for retinal detachment. AB - Retinal detachment surgery in human patients is currently 90% successful, with most detachments amenable to treatment by scleral buckling procedures. The main obstacle to achieving comparable results in veterinary patients is the active nature of our patients during the postoperative convalescent period. Adapting current techniques to include short-term chorioretinal adhesion by way of tacking, cyanoacrylate adhesives, or other methods has shown substantial promise and should be further investigated in veterinary species. The technology and methods are currently available to produce success rates comparable to those achieved in human patients, and the near future promises to bring further refinements in veterinary applications. PMID- 9326974 TI - Surgery of the orbit. AB - Orbital surgery is performed infrequently but when necessary, requires detailed understanding of orbital anatomy and the probable biologic behavior and extent of the pathologic process affecting the orbit. Thorough preoperative characterization of an orbital disease allows the surgeon to develop a surgical strategy. Inaccurate or hasty preoperative localization, determination of extension, and diagnosis may result in selection of an inappropriate surgical approach or discretionary surgery when medical treatment is indicated. In most instances, diagnostic images (MR, CT, echography) should always be made and fine needle aspiration be done before orbital surgery is performed. The choice of surgical approach or combination of approaches is determined primarily by the type, location, size, and extent of disease present. Extensive surgical exposure of the orbit is limited to centimeters or fractions of a centimeter because of the compact anatomy and tight confines of the orbital region. Careful tissue manipulation, surgical dissection, and postoperative assessment are necessary to preserve the globe and functional vision when orbital disease endangers function. PMID- 9326975 TI - Routine coronary angiography after heart transplantation. PMID- 9326976 TI - Joseph Leopold Auenbrugger (1722-1809). PMID- 9326977 TI - Surveillance cardiac catheterisation in heart transplant recipients. PMID- 9326978 TI - Myocardial supply and demand. PMID- 9326979 TI - Is blood donation good for the donor? PMID- 9326980 TI - Fax machines for thrombolysis? PMID- 9326981 TI - Clinical nurse specialists in the catheter laboratory: a time for change or a bridge too far? PMID- 9326982 TI - The phenotype/genotype relation and the current status of genetic screening in hypertrophic cardiomyopathy, Marfan syndrome, and the long QT syndrome. PMID- 9326983 TI - Myocardial oxygen supply:demand ratio as reference for coronary vasodilatory drug effects in humans. AB - OBJECTIVE: Introduction and measurement of human myocardial oxygen supply:demand ratio as a reference for quantification of coronary microvascular vasodilating drug effects in clinical studies. Myocardial oxygen consumption is the major determinant of coronary blood flow; therefore, the true vasodilating properties of coronary vasodilating drugs that may have an effect on oxygen consumption cannot be correctly assessed from blood flow changes alone. DESIGN: Prospective, controlled trial. SETTING: Academic hospital. PATIENTS: 12 patients with multivessel coronary artery disease (CAD) undergoing coronary artery bypass grafting. INTERVENTIONS: Cardiac pacing at 30 beats/min above sinus rhythm in awake and anaesthetised patients (fentanyl/pancuronium bromide). MAIN OUTCOME MEASURES: Myocardial oxygen supply, defined as coronary sinus blood flow multiplied by arterial oxygen content; myocardial oxygen demand, defined as coronary sinus blood flow multiplied by arteriovenous oxygen content difference. The change in oxygen demand induced by pacing was related to the change in myocardial oxygen supply in awake and anaesthetised patients. This myocardial oxygen supply:demand ratio determined in the reference study was compared with that induced by intravenous and intracoronary drugs (nifedipine, felodipine, urapidil, and sodium nitroprusside) in two pharmacological studies: patients with CAD undergoing cardiac surgery (45 treated with sodium nitroprusside, 27 with nifedipine, and 27 with urapidil to manage arterial blood pressure); and patients with unstable angina (and a similar degree of CAD) undergoing cardiac catheterisation for diagnostic purposes (10 treated with intracoronary nifedipine and 10 with intracoronary felodipine). RESULTS: When awake, the ratio of pacing induced oxygen supply:demand changes in the 12 reference study patients was 1.50 (95% confidence intervals (CI), 1.41-1.58), similar to the 1.45 (1.35-1.56) measured in the same patients after induction of anaesthesia. Anaesthesia per se did not increase coronary oxygen supply above the expected increase related to demand changes. The only significant change in the oxygen supply:demand ratio was induced by intracoronary bolus administration of nifedipine and felodipine (10.6 (SE 1.9) and 13.9 (1.9) ml/min, respectively, above the demand related supply). CONCLUSIONS: Quantification of coronary vasoactive properties in relation to the physiological reference ratio between myocardial oxygen supply and demand may be a powerful tool to differentiate between true and apparent coronary vasoactive drugs. PMID- 9326984 TI - Comparison of transthoracic three dimensional echocardiography with magnetic resonance imaging in the assessment of right ventricular volume and mass. AB - OBJECTIVE: Assessment of right ventricular volume and mass with three dimensional echocardiography and comparison with magnetic resonance imaging. METHODS: Measurements of right ventricular volumes performed on three dimensional datasets acquired by transthoracic echocardiography were compared to those obtained from magnetic resonance imaging performed on the same day. Volumes were measured in end systole and end diastole and ejection fraction calculated. Right ventricular mass was assessed in end systole. With both methods, the areas of a 2 mm thick slice of the ventricle were manually outlined and multiplied by the slice thickness to obtain slice volume. Slice volumes were multiplied by the number of measured slices to obtain the ventricular volume. PATIENTS: 16 patients were studied: three with normal hearts, three after surgical repair of coarctation of the aorta, nine following repair of tetralogy of Fallot, and one with Mustard atrial repair of complete transposition of the great arteries. RESULTS: Correlation between end diastolic volumes measured by both methods was r = 0.95 with limits of agreement ranging from -3.5 to 12.5 ml; correlation for end systolic volumes was r = 0.87 with limits of agreement between -4.0 and 16.4 ml; correlation for end systolic right ventricular mass was r = 0.81 with limits of agreement between -7.0 and 20.6 g. Interobserver variability ranged from 4.3% (range 0.2% to 9.3%) for end diastolic volume to 7.6% (1.8% to 15.4%) for mass measurements. CONCLUSIONS: With transthoracic three dimensional echocardiography, end diastolic right ventricular volumes can be assessed with acceptable accuracy in normal hearts and those with enlarged right ventricles, whereas the current method of three dimensional echocardiography is less good for end systolic volumes and not satisfactory for right ventricular mass measurements. PMID- 9326985 TI - Serum Lp(a) lipoprotein concentration is not associated with clinical and angiographic outcome five years after coronary artery bypass graft surgery. AB - OBJECTIVE: To examine the association between serum Lp(a) lipoprotein concentration and clinical and angiographic outcomes five years after coronary artery bypass graft (CABG) surgery. SETTING: A regional cardiothoracic centre, Freeman Hospital, and the University Clinical Investigation Unit, Royal Victoria Infirmary, Newcastle upon Tyne. PATIENTS AND DESIGN: 353 consecutive patients (56 female, 297 male, mean age 57-2 years) undergoing first time CABG surgery for stable angina were studied prospectively. MAIN OUTCOME MEASURES: Late cardiac death (beyond 30 days) and non-fatal myocardial infarction; prevalence of angina five years after surgery in 291 (94%) survivors and vein graft patency (evaluated by patient) in 118 survivors five years after surgery. Serum Lp(a) concentration and lipid profiles were measured before operation, and 3, 6, 12, and 60 months after surgery. Lipid profiles were also measured 24 months after surgery. RESULTS: Weighted Lp(a) concentration (by tertile) was not associated with late cardiac death or with the combination of late cardiac death and non-fatal myocardial infarction, with the presence of angina, or with vein graft occlusion. The association remained non-significant if analysis was restricted to the upper tertile of LDL cholesterol (> 4.1 mmol/l) or to patients under the age of 55 years at the time of surgery. CONCLUSIONS: Serum Lp(a) concentration did not predict late cardiac death, the combination of late cardiac death and non-fatal myocardial infarction, or the prevalence of angina or vein graft occlusion five years after CABG surgery. PMID- 9326986 TI - The natural history of aneurysmal coronary artery disease. AB - OBJECTIVE: To assess the contribution of coronary artery ectasia, either isolated or in association with obstructive coronary artery disease, to morbidity and mortality from ischaemic heart disease. DESIGN: A retrospective study of patients undergoing coronary arteriography at a tertiary cardiac centre. PATIENTS AND METHODS: The epidemiological, clinical, arteriographic, and follow up characteristics of three groups of patients were examined: group A, 172 patients with coronary artery ectasia and coexisting significant coronary artery disease; group B, 31 patients with coronary artery ectasia only; group C, 165 patients with significant coronary artery disease but without ectasia, matched for sex and age with group A. RESULTS: Group A patients had a similar incidence of a previous myocardial infarction to group C patients (61.6% v 64.2%), exercise performance, severity of obstructive lesions (CASS score 2.19 v 2.14), and similar distribution of diseased vessels. At follow up of approximately two years they experienced a similar incidence of unstable angina (7.5% v 4.4%) and myocardial infarction plus cardiac death (4.9% v 6.1%). They underwent bypass surgery with similar frequency (39% v 42%) but there was a lower frequency of percutaneous transluminal coronary angioplasty (5.8% v 17%, P < 0.01). Patients with pure coronary ectasia (group B) had a lower incidence of a previous myocardial infarction (38.7%, 12/31, P < 0.05) than the two other groups. The infarct in all cases was related to an ectatic artery. Their exercise performance and ejection fraction (9 (SD 3) minutes and 56.5(9)%) were higher (P < 0.01) than group A (5 (2) minutes, 48.3(10)%) and group C (5.3 (2) minutes, 49.3(10)%). Group B had no myocardial infarctions, cardiac death, surgery, or intervention at follow up; 4.4% (5/115) developed unstable angina. The incidence of angina at study entry was similar in all three groups (38.7-49.7%). CONCLUSIONS: Coronary artery ectasia does not confer added risk in patients with coexisting obstructive coronary artery disease. Although there is a measurable incidence of previous myocardial infarction, patients with pure ectasia have a good prognosis. The wisdom of giving oral anticoagulants to such patients is questioned. PMID- 9326987 TI - Comparative study of chest pain characteristics in patients with normal and abnormal coronary angiograms. AB - OBJECTIVE: To improve the characterisation of chest pain by comparing symptoms in patients with normal and abnormal coronary angiograms. STUDY DESIGN: Prospective case-control study. SETTING: Single tertiary cardiac referral centre. PATIENTS: 65 consecutive patients with chest pain and completely normal coronary angiograms recruited over a period of one year, and 65 sex matched patients with significant stenoses at angiography. MAIN OUTCOME MEASURES: Standardised chest pain questionnaires. RESULTS: 61 of 65 patients (94%) and every control reported chest pain on exertion. There were no important differences in the site, quality, and radiation of pain but three symptoms had discriminatory value expressed in binary fashion ("typical" v "atypical"): the consistency with which pain was reproduced by exercise (typical, score index 10/10), the duration of pain episodes (typical, five minutes), and the frequency of pain at rest (typical, 10% all pain episodes). All three symptoms were atypical in 21 (32%) patients with normal coronary angiograms, but only one patient with an abnormal coronary angiogram. Patients with no typical features had a 2% chance of an abnormal coronary angiogram if aged under 55 years or 12% if aged 55 years or more. The additional impact of exercise stress testing was low. CONCLUSIONS: Chest pain characteristics which separate patients with normal coronary angiograms from patients with obstructive coronary heart disease can be defined objectively. This may allow improvements in referral patterns for specialist opinion or angiography, and in characterisation of patients in research studies. PMID- 9326988 TI - Serum urate and the risk of major coronary heart disease events. AB - OBJECTIVE: To examine the relation between serum urate and the risk of major coronary heart disease events. DESIGN: A prospective study of a male cohort. SETTING: One general practice in each of 24 British towns. SUBJECTS: 7688 men aged 40-59 years at screening. MAIN OUTCOME MEASURES: Fatal and non-fatal coronary heart disease events. RESULTS: There were 1085 major coronary heart disease events during the average follow up period of 16.8 years. Serum urate was significantly associated with a wide range of cardiovascular risk factors including body mass index, alcohol intake, antihypertensive treatment, pre existing coronary heart disease, serum triglycerides, cholesterol, and diastolic blood pressure. There was a significant positive association between serum urate and risk of coronary heart disease after adjustment for lifestyle factors and disease indicators. This relation was attenuated to non-significance upon additional adjustment for diastolic blood pressure and serum total cholesterol: cholesterol appeared to be the critical factor in attenuating this relation. When the association between serum urate and risk of coronary heart disease was examined by presence and grade of pre-existing coronary heart disease, a positive association was seen only in men with previous definite myocardial infarction, even after full adjustment (P = 0.07). CONCLUSIONS: The relation between serum urate and the risk of coronary heart disease depends heavily upon the presence of pre-existing myocardial infarction and widespread underlying atherosclerosis as well as the clustering of risk factors. Thus serum urate is not a truly independent risk factor for coronary heart disease. Raised serum urate appears to be an integral part of the cluster of risk factors associated with the insulin resistance syndrome that include obesity, raised serum triglycerides, and serum cholesterol. PMID- 9326989 TI - Sequelae after modified Fontan operation: postoperative haemodynamic data and organ function. AB - OBJECTIVE: To investigate the specific sequelae of the Fontan operation, and particularly the potential sequelae of chronically elevated systemic venous pressure. DESIGN: A retrospective analysis of clinical and haemodynamic data and evaluation of organ function in 80 surviving patients undergoing modified Fontan operation for various forms of underlying functionally univentricular hearts. PATIENTS: 65 patients (81%) who had undergone a total cavopulmonary anastomosis and 15 an atriopulmonary anastomosis. Follow up ranged from 12 to 106 months (mean 54 (SD 23) months). RESULTS: 62 patients underwent postoperative cardiac catheterisation (mean systemic venous pressure 10.5 (2.5) mm Hg and cardiac index 3.1 (0.7) l/min/m2). Older age at operation was significantly correlated with both higher systemic venous pressure and lower cardiac index. Atrial arrhythmia was documented on Holter electrocardiogram in 17%. Protein losing enteropathy (with abnormal alpha 1-antitrypsin clearance) was found in 2/80 patients (2.5%). Ten patients had hypoproteinaemia, with a significantly higher incidence in patients after total cavopulmonary anastomosis and young age at operation. Liver function tests reflecting liver synthesis and metabolism were normal in all, whereas mild cholestasis was found in nearly 30%-predominantly in patients with a cardiac index of < 3 l/min/m2 (P = 0.045). Five patients (6.2%) developed atrial thrombosis. Coagulation factor analysis in 44 patients showed protein C deficiency in 11 (25%); laboratory signs of activation of the coagulation system were found in four of these (9%). None of the abnormal laboratory indices was significantly related to underlying cardiac malformation, postoperative systemic venous pressure, or follow up interval. CONCLUSIONS: A high proportion of clinically asymptomatic patients had abnormal laboratory findings on mid-term follow up. Detailed evaluation of organ function is necessary to detect the need for further diagnostic procedures before clinical symptoms develop. PMID- 9326990 TI - Radiofrequency catheter ablation of accessory pathways in infants. AB - OBJECTIVE: To evaluate the indications, results and complications of radiofrequency catheter ablation in small infants with supraventricular tachycardia due to an accessory atrioventricular pathway. METHODS: Five infants less than 9 months old underwent radiofrequency catheter ablation of accessory pathways. Ablation was done for medically refractory tachyarrhythmia associated with aborted sudden death in two patients, left ventricular dysfunction in one, failure of antiarrhythmic drugs in one, and planned cardiac surgery in one. All five patients underwent a single successful procedure. Three left free wall pathways were ablated by transseptal approach, a right posteroseptal pathway was ablated from the inferior vena cava, and a left posteroseptal pathway was approached from the inferior vena cava into the coronary sinus. A deflectable 5F bipolar electrode catheter with a 3 mm tip was used. RESULTS: A sudden increment in impedance indicative of coagulum formation was observed in two procedures. One patient developed a transient ischaemic complication after ablation of a left lateral accessory pathway by transseptal approach. This patient had mild pericardial effusion after the procedure. Moderate pericardial effusion was also noted in another patient. After a mean follow up of 18.4 months all patients are symptom free without treatment. CONCLUSIONS: Radiofrequency catheter ablation can be performed successfully in infants. Temperature monitoring in 5F ablation catheters would be desirable to prevent the development of coagulum. Echocardiography must be performed after the ablation procedure to investigate pericardial effusion. PMID- 9326991 TI - Late ventricular potentials and heavy drinking. AB - OBJECTIVES: To assess the effects of chronic drinking on detection of low amplitude signals, and to determine the relation between late ventricular potentials (LVP) and liver biopsy findings. DESIGN: Prospective study. SETTING: General hospital. PATIENTS: 41 consecutive chronic alcoholics without known pre existing heart disease. METHODS: About four days after each patient's last alcoholic drink, ECG, echocardiography, signal averaged electrocardiogram, liver biopsy, and blood tests were performed. RESULTS: Twenty eight per cent of patients had evidence of LVP. There was a correlation between the percentage of steatosis of the hepatic biopsy and the amplitude of the last 40 ms of average QRS (P = 0.04), the duration of the terminal low amplitude QRS signal (P = 0.05), and the number of positive criteria of late potentials (P = 0.02). CONCLUSIONS: Chronic drinking sufficient to cause steatosis is associated with positive findings on the signal averaged ECG. PMID- 9326992 TI - Effect of phenylephrine infusion on atrial electrophysiological properties. AB - OBJECTIVE: To determine the effect of changes in autonomic tone induced by phenylephrine infusion on atrial refractoriness and conduction. DESIGN: Left and right atrial electrophysiological properties were measured before and after a constant phenylephrine infusion designed to increase sinus cycle length by 25%. SUBJECTS: 20 patients, aged 53 (SD 6) years, undergoing electrophysiological study for investigation of idiopathic paroxysmal atrial fibrillation (seven patients) or for routine follow up after successful catheter ablation of supraventricular tachycardia (13 patients). MAIN OUTCOME MEASURES: Changes in left and right atrial effective refractory periods, atrial activation times, and frequency of induction of atrial fibrillation. RESULTS: Phenylephrine (mean dose 69 (SD 18) mg/min) increased mean blood pressure by 22 (12) mm Hg (range 7 to 44) and lengthened sinus cycle length by 223 (94) ms (20 to 430). Left atrial effective refractory period lengthened following phenylephrine infusion from 250 (25) to 264 (21) ms (P < 0.001) but there was no significant change in right atrial effective refractory period: 200 (20) v 206 (29), P = 0.11. There was a significant relation between the effect of phenylephrine on sinus cycle length and on right atrial refractoriness (r = 0.6, P = 0.005) with shortening of right atrial refractoriness in patients with the greatest prolongation in sinus cycle length. During phenylephrine infusion, the right atrial stimulus to left atrial activation time at the basic pacing cycle length of 600 ms was unchanged, at 130 (18) v 131 (17) ms, but activation delay with a premature extrastimulus increased: 212 (28) v 227 (38) ms, P = 0.002. Atrial fibrillation was induced by two of 58 refractory period measurements at baseline and by 12 of 61 measurements during phenylephrine infusion (P < 0.01). Phenylephrine increased the difference between left and right atrial refractory periods by 22.8 (19.4) ms in the five patients with induced atrial fibrillation after phenylephrine compared to 0.9 (16.2) ms in the 13 patients without induced atrial fibrillation after phenylephrine infusion (P = 0.02). CONCLUSIONS: Phenylephrine infusion increased left atrial refractoriness and intra-atrial conduction delay following a premature right atrial extrastimulus. Induction of atrial fibrillation during phenylephrine infusion was associated with non-uniform changes in atrial refractoriness. These data support the concept that changes in autonomic tone may precipitate atrial fibrillation in susceptible individuals. PMID- 9326993 TI - Comparison of bronchopulmonary collaterals and collateral blood flow in patients with chronic thromboembolic and primary pulmonary hypertension. AB - OBJECTIVE: To compare the visualisation of bronchopulmonary collaterals and bronchopulmonary collateral blood flow in patients with chronic thromboembolic pulmonary hypertension 2nd primary pulmonary hypertension. SETTING: Referral centre for cardiology at an academic hospital. PATIENTS: Nine patients with chronic thromboembolic pulmonary hypertension and 17 with primary pulmonary hypertension. INTERVENTIONS: Bronchopulmonary collaterals were visualised by selective bronchial arteriography or thoracic aortography. Bronchopulmonary collateral blood flow was estimated by injecting indocyanine green into the ascending aorta and sampling below the mitral valve from the left ventricle. RESULTS: The degree of pulmonary hypertension was comparable in the two groups. Large bronchopulmonary collaterals were visualised in all the patients with thromboembolic pulmonary hypertension who had bronchial arteriography or aortography or both. None of the primary pulmonary hypertension group studied by aortography had bronchopulmonary collaterals (P < < 0.001). All the patients with chronic thromboembolic pulmonary hypertension had significant bronchopulmonary collateral blood flow, which was (mean (SD)) 29.8 (18.6)% of the systemic blood flow. There was no recordable collateral blood flow in 11 of 15 patients with primary pulmonary hypertension. In the remaining four patients the mean value was 1.1 (1.8)% of the systemic blood flow (P < < 0.001). CONCLUSIONS: Visualisation of bronchopulmonary collaterals by thoracic aortography or by bronchial arteriography, or the demonstration of an increased bronchopulmonary collateral flow, helps to distinguish patients with chronic thromboembolic pulmonary hypertension from those with primary pulmonary hypertension. PMID- 9326994 TI - Abnormal skeletal muscle bioenergetics in familial hypertrophic cardiomyopathy. AB - OBJECTIVE: To determine the skeletal muscle metabolic manifestations of familial hypertrophic cardiomyopathy. DESIGN: A case-control study. SETTING: 31P magnetic resonance spectroscopy of the calf muscle was performed on volunteers from a centre specialising in familial hypertrophic cardiomyopathy. PATIENTS: Five patients with abnormal beta myosin heavy chain protein in cardiac and skeletal muscle and five patients with a troponin T abnormality in cardiac muscle were compared with healthy controls. RESULTS: High energy phosphate metabolism in vivo was examined in a non-invasive manner. In resting muscle, the beta myosin heavy chain group had a higher ratio of phosphocreatine to ATP concentration (4.51 (SD 0.17)) than either the troponin T group (3.88 (0.42)) or controls (n = 16; 4.04 (0.40)). Exercise duration was reduced compared to controls, and during the fourth minute of exercise phosphocreatine depletion and muscle acidification were greater in both patient groups. After exercise, the recovery of phosphocreatine an index of oxidative metabolic capacity of the muscle-was slower in the beta myosin heavy chain group (mean half time 0.65 (0.08) minutes) than in the troponin T group (0.60 (0.17) minutes) or controls (0.48 (0.14) minutes). CONCLUSIONS: Exercise metabolism was abnormal in both groups of subjects, and the affected contractile protein determined the metabolic changes in muscle at rest and during recovery. In patients with abnormal beta myosin heavy chain protein, there was a decrease in oxidative capacity consistent with the reduction in mitochondria reported in muscle biopsy studies of similar patients. PMID- 9326995 TI - Health related quality of life and psychological wellbeing in patients with hypertrophic cardiomyopathy. AB - OBJECTIVE: To assess the health related quality of life and psychological wellbeing of patients with hypertrophic cardiomyopathy, to correlate these with symptoms, clinical, and psychosocial factors. DESIGN: Questionnaire distributed to 171 hypertrophic cardiomyopathy patients aged at least 14 years, selected at random from a dataset of 480 patients. Assessments included the Short Form 36 (SF 36) Health Survey, the Hospital Anxiety and Depression questionnaire, and measures of adjustment, worry, and patient satisfaction. RESULTS: There was an 80.1% response rate to the questionnaire. Patients had severe limitations in all eight dimensions of quality of life assessed by the SF-36: physical functioning, role limitations owing to physical problems, role limitations owing to emotional problems, social functioning, mental health, general health perceptions, vitality, and bodily pain. Levels of anxiety and depression were also high compared with population norms. Quality of life was particularly impaired in patients with chest pain and dyspnoea, but was less consistently related to clinical cardiological measures. Adjustment to the condition and patient satisfaction were generally good. In multivariate analysis, quality of life was associated with a combination of symptom patterns and psychosocial factors. No differences in quality of life, anxiety or depression were observed between patients with no known family history, those with familial cardiomyopathy, and patients with a family history of premature sudden death. CONCLUSIONS: Hypertrophic cardiomyopathy is associated with substantial restrictions in health related quality of life. Symptoms, adjustment, and quality of interactions with clinical staff contribute to these limitations. Recognition of the problems confronted by patients with hypertrophic cardiomyopathy requires continued efforts at education both of the public and health professionals. PMID- 9326997 TI - Cardiac catheterisation performed by a clinical nurse specialist. AB - OBJECTIVE: To establish the feasibility and safety of an appropriately trained clinical nurse specialist performing diagnostic cardiac catheterisation. DESIGN: Non-randomised retrospective comparison between the first 100 and second 100 consecutive investigations by a clinical nurse specialist and 200 consecutive patients investigated by two cardiology registrars over a similar period. SETTING: Regional cardiac centre performing 3200 catheterisation procedures per annum. PATIENTS: 200 patients undergoing routine (non-emergency) cardiac catheterisation for investigation of ischaemic heart disease. MAIN OUTCOME MEASURES: Procedural complications, image quality, fluoroscopy times. RESULTS: Satisfactory diagnostic images in all nurse specialist cases with no deaths and two complications (coronary artery dissection and femoral pseudoaneurysm). Procedure duration and fluoroscopy times slightly shorter for clinical nurse specialist by 3 and 1.6 minutes, respectively (P < 0.05). CONCLUSIONS: Non medical practitioners can be trained to perform straightforward cardiac angiography in low risk patients with consultant supervision, as for cardiology registrars. With important restrictions such posts may have a limited role in supporting future consultant based services. PMID- 9326996 TI - Possible association of a reduction in cardiovascular events with blood donation. AB - BACKGROUND: The iron hypothesis suggests that females are protected from atherosclerosis by having lower iron stores than men, thus limiting oxidation of lipids. OBJECTIVE: To test the iron hypothesis by comparing cardiovascular event rates in whole blood donors compared with nondonors. DESIGN: Prospective cohort with telephone survey follow up. SETTING: The State of Nebraska, USA. PARTICIPANTS: A sample was selected from the Nebraska Diet Heart Survey (NDHS) restricting for age > or = 40 years and absence of clinically apparent vascular diseases at time of enrollment in to NDHS (1985-87). MAIN OUTCOME MEASURES: The occurrence of cardiovascular events (myocardial infarction, angina, stroke), procedures (angioplasty, bypass surgery, claudication, endarterectomy), nitroglycerin use, or death (all cause mortality), and level of blood donation. RESULTS: Participants were 655 blood donors and 3200 non-donors who differed in education, physical activity, diabetes, and frequency of antihypertensive treatment; 889 were lost to follow up. Sixty four donors and 567 non-donors reported cardiovascular events (crude odds ratio = 0.50, 95% confidence interval (CI) 0.38-0.66). The benefit of donation was confined to non-smoking males (adjusted odds ratio 0.67, 95% CI 0.45-0.99). Benefit was limited to current donors (the most recent three years). No additional benefit resulted from donating more than once or twice over three years. CONCLUSION: In support of the iron hypothesis, blood donation in non-smoking men in this cohort was associated with reduced risk of cardiovascular events. A randomised clinical trial is warranted to confirm these findings as the observed personal health benefit of donation has public policy ramifications. PMID- 9326999 TI - Images in cardiology. Echocardiographic appearances of right ventricular dysplasia in the fetus. PMID- 9326998 TI - Use of fax facility improves decision making regarding thrombolysis in acute myocardial infarction. AB - BACKGROUND: Electrocardiography is the fundamental investigation for decision making regarding thrombolytic treatment in acute myocardial infarction (MI). Increasing the accuracy of ECG analysis by input from consultant staff may assist in management decisions in patients with suspected MI. AIMS: To evaluate a system whereby out of hours ECGs can be faxed to the consultant to aid in decision making regarding thrombolytic treatment. METHODS: 112 patients with suspected MI were assessed on admission by the senior house officer (SHO) who faxed to a cardiology consultant the ECG trace and a predesigned form with information on: clinical assessment of the patient; interpretation of the ECG; and views regarding administration of thrombolytic treatment including choice of agent. The consultant reviewed the information and communicated his views to the SHO. Subsequent diagnosis was recorded in all patients and the forms were analysed in regard to areas of agreement and disagreement between the SHO and the consultant. RESULTS: A diagnosis of MI was confirmed in 52 of the 112 patients (46.4%). The consultant agreed with the SHO's decision on thrombolysis in 98 patients (87.5%). The reason for disagreement in the remaining 14 patients (12.5%) was SHO misinterpretation of the ECG (10 patients) and clinical assessment (four patients). Eight patients were saved unnecessary thrombolytic treatment and four received it when they otherwise would not have. Additionally the choice of thrombolytic agent was changed in six patients from streptokinase to tissue plasminogen activator. CONCLUSION: The use of fax machine assists in decision making with regard to thrombolytic treatment and provides support to junior doctors in what can be a difficult, yet critical decision. PMID- 9327000 TI - Simultaneous delivery of two patent arterial duct coils via one venous sheath. AB - A female child, 10 months of age, weighing 7.2 kg, was catheterised for closure of a patent arterial duct. Aortography was performed in the lateral projection and the minimum diameter of the arterial duct was assessed by comparing it to the size of the catheter. The duct size was estimated between 3 and 3.5 mm at the narrowest point, therefore, it was decided to deliver two 5 mm patent arterial duct coils to avoid placement of an 8 mm coil in this small child. Similar operations were subsequently performed in two further children. Simultaneous delivery of two coils via a single long venous sheath is easy, fast, and safe. This simple and inexpensive procedure can reduce irradiation and anaesthesia time. PMID- 9327001 TI - Transcatheter embolisation of an enlarging acquired coronary arteriovenous fistula in a heart transplant recipient. AB - A 50 year old, recent cardiac transplant recipient developed systolic and diastolic murmurs but remained asymptomatic. The cause of the murmurs was not evident at transthoracic echocardiography. During routine left heart catheterisation a left anterior descending artery to right ventricular fistula was evident arising from the distal vessel and presumably acquired during routine endomyocardial biopsy. One year later, the patient remained asymptomatic but the calibre of the left anterior descending artery had increased and there appeared to be poor flow in to the proximal branches. The fistula was successfully treated by percutaneous deployment of two detachable embolisation coils in to the distal left anterior descending artery. PMID- 9327002 TI - Single coronary artery with infundibular pulmonary stenosis. PMID- 9327003 TI - WHO and its role in the prevention of deafness and hearing impairment. PMID- 9327004 TI - Long-term observation of subcutaneous tissue reaction to synthetic auditory ossicle (Apaceram) in rats. AB - The present study evaluates histological characteristics of the soft tissue response to long-term implantation of Apaceram discs composed of dense hydroxyapatite in rats. Discs were implanted into the subcutaneous tissue of 76 rats for six to 20 months. Decalcified histological sections stained with haematoxylin and eosin (H & E) and Mallory's azan were examined. Different cell types surrounding implants were counted. The greatest proportion of macrophages was found at six months (13.5 per cent). This proportion gradually decreased to four per cent at 20 months. Small numbers of lymphocytes and foreign body giant cells were observed in every group, but neither neutrophils nor osteogenesis were observed in any specimens. Results of the present study and previous related studies indicate that despite reappearance of a small number of macrophages six months after implantation, Apaceram is useful for reconstructive surgery. PMID- 9327005 TI - Temporalis fascia grafts shrink. AB - Temporalis fascia, placed as an underlay graft, is commonly used to repair tympanic membrane perforations. Graft failure, however, is a well recognized complication. Grafts are often allowed to dry out during the procedure and, therefore, are often positioned in a dry or partially dehydrated state and only become fully rehydrated after placement. This study looked at how the size of the temporalis fascia alters with its state of hydration. The size of 20 temporalis fascia grafts was measured when fresh, after flattening and allowing them to dry, and finally after rehydrating the grafts with 0.9 per cent saline solution. Significant shrinkage was demonstrated. It is therefore proposed that a cause of increased failure rates, particularly in anterior myringoplasties, is loss of underlay due to graft rehydration and shrinkage. Thus graft shrinkage should be considered when positioning the graft. PMID- 9327006 TI - Primary tumours of the vestibule and inner ear. AB - Seven primary tumours of the vestibule and inner ear are described, six schwannomas and one traumatic fibroma. Schwannomas in this situation may occur as sporadic tumours, or may be a feature of neurofibromatosis type 2 (NF-2). In the latter condition they may occur in isolation or in association with, but separate from, schwannomas arising in the internal meatus. Direct extension into the vestibule of an intrameatal vestibular schwannoma is well reported, but extension of an intravestibular tumour into the internal meatus is not described. Traumatic fibromas of the vestibule are rare and the trigger could be an attack of labyrinthitis. Intravestibular tumours, although rare, are likely to be diagnosed with increasing frequency with the widespread use of MR imaging. PMID- 9327007 TI - Intra-operative facial nerve monitoring. Its predictive value after skull base surgery. AB - PURPOSE: Facial nerve monitoring can be used to predict post-operative facial function after skull base surgery. In this study three methods of prediction of facial function were compared. These methods utilize various parameters of the evoked electromyographic monitoring. MATERIAL AND METHODS: Twenty-three patients who underwent surgery for skull base diseases were retrospectively reviewed. Amplitude of ongoing electromyographic activity, stimulation current thresholds and amplitude of evoked response were analysed. The predictive value of the three methods was correlated with post-operative facial nerve function. RESULTS: The method that used only the stimulation thresholds predicted the final post operative facial function in 86.9 per cent of the patients. The second employed a mathematical ratio which combined the amplitude of evoked response and the stimulation current thresholds and confirmed the prediction of the facial function in 91.3 per cent of the patients. The last method does not consider the stimulation thresholds greater than 0.05 mA and failed to predict the final VIIth nerve function in patients in whom the stimulation was greater than 0.05 mA. CONCLUSION: Analysis of prognostic value demonstrates that the first two methods had the smaller degree of variation showing the better sensitivity. PMID- 9327008 TI - The Groote Schuur hospital classification of the orbital complications of sinusitis. AB - The complications of sinusitis have been well described. The most common classifications used for orbital complications have been that of Chandler et al. (1970) and Moloney et al. (1987). With the ready availability of high-resolution computed tomography (CT) scanners, limitations of these classifications have become apparent. The aims of this study were to determine the relative frequency of the various complications associated with acute sinusitis, to determine which groups of sinuses were most frequently involved and to correlate the orbital signs with a new proposed classification of orbital complications. Over a five year period, 87 consecutive patients were admitted with acute sinusitis. Sixty three patients (72.4 per cent) had one or more complications. When orbital complications were classified under the proposed classification, all patients with proptosis and/or decreased eye movement had post-septal infection. Visual impairment occurred only in the post-septal group. Most complications had a combination of sinus involvement with the maxillary/ethmoid/frontal combination being the most common. The authors propose a modification of Moloney's classification for orbital complications of acute sinusitis that allows a clear differentiation between pre- and post-septal infection and a radiological differentiation to be made between cellulitis/phlegmon and abscess formation. The latter is of importance when a decision is made on whether surgical intervention is appropriate or not. PMID- 9327009 TI - Nasopharyngeal carcinoma: clinical trends. AB - Nasopharyngeal carcinoma (NPC) is one of the most difficult diseases to diagnose at an early stage. The clinical presentation of 122 patients with confirmed NPC is described and the findings analysed. The common modes of presentation and cases where detailed nasopharyngeal examination need to be performed are highlighted. We emphasize the importance of health education and training for primary care physicians for early detection of these cases. PMID- 9327010 TI - Supraglottic laryngectomy--series report and analysis of results. AB - Between October 1987 and October 1993, 92 patients with squamous cell carcinoma of the supraglottis were treated by supraglottic laryngectomy and neck dissection in our department. There were 33 T1, 46 T2, six T3 and seven T4 cases. All patients with N+ necks and T3 or T4 tumours received post-operative radiotherapy (5,000-6,500 cGy). The patients were followed for a minimum of 36 months or until death. The incidence of local recurrence was 7.6 per cent. Neck recurrence was observed in 13 per cent of patients. Decannulation was achieved in 93.4 per cent of the cases with three patients undergoing gastrostomy because of aspiration. The average hospital stay was 26 days. The overall three-year survival was 83.6 per cent, with eight patients dying of unrelated causes. There was a significant difference in recurrence rate between patients in the N0 and the N+ stage. PMID- 9327011 TI - Diathermy tonsillectomy: comparisons of morbidity following bipolar and monopolar microdissection needle excision. AB - Tonsillectomy is frequently associated with a considerable post-operative morbidity. In some cases reactionary or secondary haemorrhage occurs and all patients suffer a degree of post-operative pain. The use of bipolar diathermy excision has become popular because it reduces intra-operative blood loss, but all diathermy inevitably produces a degree of damage to adjacent normal soft tissues. In turn this inadvertent injury must act to increase the post-operative pain. Monopolar dissection using a fine tungsten diathermy needle (the Colorado needle) allows sharp dissection at low power levels and in previous studies has been shown to produce a reduction in collateral tissue damage. In this prospective study the morbidity associated with tonsillectomy using this needle was compared to that following a standard bipolar dissection. Using the monopolar needle produced no enhanced risk of reactionary or secondary haemorrhage while causing significantly less post-operative pain and a reduction in eschar. We believe that excision using this needle preserves the advantages associated with bipolar dissection while reducing local soft tissue damage. PMID- 9327012 TI - The evaluation of velopharyngeal function using flexible nasendoscopy. AB - Nasendoscopy is an essential tool in assessing the dynamic function and structure of the velopharyngeal sphincter during speech and swallowing. Flexible fibre optic nasendoscopy has been used by the cleft palate team at Withington Hospital, Manchester since 1989. Seventy-six patients were referred between 1989 and 1994 for evaluation of velopharyngeal function during speech. Flexible nasendoscopic evaluation was attempted in 50 patients, and successfully carried out in 43 patients. The age range was four years to 77 years (mean 21 years). The patients were divided into two groups: Group 1 consisting of patients with cleft palate and Group 2 comprised of patients with non-overt cleft palate-related velopharyngeal dysfunction of various aetiologies; such as, submucous cleft, post tonsillectomy, post-adenoidectomy, neurological and post-traumatic. Based on the findings on nasendoscopy, videofluoroscopy and clinical speech/voice analysis the following treatment options were recommended: 17 (40 per cent) for pharyngoplasty, five (11 per cent) for revision pharyngoplasty, 15 (35 per cent) for speech therapy, four for an obturator and one for tonsillectomy. Two previously undetected submucous clefts were diagnosed. PMID- 9327013 TI - Misleading clinical features in Wegener's granulomatosis. A case report. AB - Wegener's granulomatosis is a systemic vasculitis that may present with a variety of findings and be difficult to diagnose. We report a case of a patient who presented with serous otitis media and subsequently developed a suspected primary lung tumour. Thoracotomy and pulmonary mass excision were required to establish the diagnosis. Otological manifestations of Wegener's granulomatosis, differential diagnosis, pathological findings and c-ANCA test role are discussed. PMID- 9327014 TI - Penetrating oral foreign body presenting as an aural polyp. AB - A case of a penetrating oral foreign body presenting with an aural polyp is described. The possibility of a penetrating oral injury should be considered whenever a child's fall is unwitnessed, as it is easily overlooked. An underlying foreign body should be considered in cases where an aural polyp fails to respond to standard therapy. MRI may be the best imaging technique to identify plastic foreign material. PMID- 9327015 TI - The role of surgery in tuberculous mastoiditis: appropriate chemotherapy is not always enough. AB - We present a case of tuberculous otitis media in which a facial palsy occurred after the start of appropriate chemotherapy. To our knowledge this circumstance has not been described previously. It has been argued that radical surgery is completely unnecessary if chemotherapy is commenced early in the disease. We would suggest that this is not always the case, and would advocate a more measured approach. PMID- 9327016 TI - Large endolymphatic sac. A congenital deformity of the inner ear shown by magnetic resonance imaging. AB - Fluctuant and progressive hearing impairment in a patient with a wide vestibular aqueduct has been called the 'large vestibular aqueduct syndrome'. Recently reports of magnetic resonance imaging (MRI) studies describe enlargement of the endolymphatic sac and duct in patients shown to have large vestibular aqueducts by computed tomography (CT). A patient with progressive deafness was shown to have borderline or slightly enlarged vestibular aqueducts by re-formatted sagittal CT. However, MRI in axial and sagittal planes gave a more satisfactory demonstration of both aqueduct and endolymphatic sac enlargement. PMID- 9327018 TI - Cilia from a cystic fibrosis patient react to the ciliotoxic Pseudomonas aeruginosa II lectin in a similar manner to normal control cilia--a case report. AB - The ciliary beat frequency measurements taken from a nasal polyp from a cystic fibrosis patient were similar to that of the control nasal polyps. The addition of a ciliotoxic lectin produced by Pseudomonas aeruginosa stopped the beating of the cilia as in the controls. This reaction could be blocked by the pre incubation of the lectin with its inhibitor fucose. As in the control, the addition of fucose after the cilia had slowed resulted in a return to normal ciliary beating within 24 hours. This shows that the delta F508 CF mutation observed in this patient does not affect ciliary beating and suggests that treatment with fucose in the early stages of a Pseudomonas aeruginosa infection could be advantageous for cystic fibrosis patients. PMID- 9327017 TI - The pathophysiology of otitic hydrocephalus. AB - The pathophysiology of otitic hydrocephalus remains controversial. It has been argued that involvement of the superior sagittal sinus, by, at least, a mural thrombus is a necessary component of this disease. We present a case of otitic hydrocephalus where on magnetic resonance imaging (MRI) normal luminal and mural flow within the superior sagittal sinus is demonstrated. The presence of thrombus in the lateral venous sinus alone appears sufficient in this case to impede venous drainage of the intracranial contents into the neck and produce a rise in the cerebral venous pressure and a subsequent increase in the CSF pressure. The presence of a superior sagittal sinus mural thrombus is not required. PMID- 9327019 TI - Laryngeal necrosis after combined chemotherapy and radiation therapy. AB - Post-radiation necrosis of the larynx is a major complication after irradiation and has become rare. Recently, combined chemotherapy and radiation therapy has been introduced for head and neck tumours. The authors report a case of laryngeal necrosis after combination therapy for a patient with cervical lymph node metastases of nasopharyngeal carcinoma and review the literature on late laryngeal necrosis. Although radiation-induced laryngeal necrosis has become a rare complication, the combination of chemotherapy and radiation therapy may increase its incidence. We should always consider it as a possible late complication and treat it appropriately. PMID- 9327020 TI - Patient with primary tonsillar and gastric syphilis. AB - A male patient with syphilitic lesions in the tonsil and stomach is presented. The patient was infected while practising oral sex with heterosexual friends. He complained of nausea and snoring; his left tonsil was enlarged. Spirochetes were detected in a smear preparation from the left tonsil. As a gastric lesion, initially believed to be cancer, appeared to result from spirochete ingestion, the case is considered to represent primary syphilis. After antibiotic therapy with ampicillin, the left tonsil returned to normal size and gastric changes were no longer evident endoscopically. Gastroscopy should be considered if syphilis of the tonsil is observed, particularly when gastrointestinal symptoms are present. Both the oral and the gastric lesion can be mistaken for malignant neoplasm. PMID- 9327021 TI - An unique tumour of the geniohyoid muscle: an intramuscular haemangioma. AB - We present the first case report in the English literature of an intramuscular haemangioma of the geniohyoid muscle. This occurred in a 24-year-old female and the diagnosis was not made prior to resection. Haemangiomas are uncommon tumours of the head and neck and intramuscular haemangiomas account for fewer than one per cent of the total. Diagnosis of the vascular nature of the tumour is often missed. Recurrence is common and usually due to incomplete excision. A review of the literature and a case report of these locally destructive lesions is presented. PMID- 9327022 TI - Aberrant common carotid artery encountered during free jejunal graft repair of oesophageal stricture. AB - The origin and course of the carotid arterial system in the superior mediastinum and neck are remarkably constant. Although variations are rare, they can have important implications in certain clinical problems. In this report, a patient underwent free jejunal graft repair of a cervical oesophageal stricture caused by radiotherapy for a post-cricoid carcinoma. The common carotid artery on the side of the surgical approach was found to cross the midline to the other side of the neck, and created difficulties both with the procedure and future management. This has not been previously reported. PMID- 9327023 TI - An unusual foreign body in the post-nasal space: a case report. AB - We report a case of a 13-year-old boy who inserted a 5 cm nail into his nose. X ray showed it in the post-nasal space and it was removed through the mouth. PMID- 9327024 TI - Tumours and pseudotumorous lesions of the temporomandibular joint: a diagnostic challenge. AB - Tumours and pseudotumorous lesions originating from the synovial membrane of the temporomandibular joint are rare. We report a series of six cases of such disorders. There were two cases of synovial chondromatosis, two of calcium pyrophosphate dihydrate crystal deposition disease, one nodular synovitis and one synovial sarcoma. Three patients were female and three were male. Their ages ranged from 36 to 70 years. All had atypical clinical and radiographical presentation. The prevalence, clinical and radiographical findings and pathological features of each disease entity are discussed and a review of the literature is made concerning all tumours and pseudotumours arising from the temporomandibular joint. PMID- 9327025 TI - Surgical aspects of paediatric cochlear implantation. PMID- 9327026 TI - Percutaneous endoscopic gastrostomies: are they being done for the right reasons? PMID- 9327027 TI - Post-transplantation lymphoproliferative disease. PMID- 9327028 TI - Altered folate and vitamin B12 metabolism in families with spina bifida offspring. AB - Folic acid intake reduces the risk of neural tube defects (NTDs). Although the 677C-->T mutation in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene is a risk factor for NTDs, it only partly explains the elevated homocysteine levels in mothers of children with NTDs. We measured vitamin B12, folate and homocysteine in patients with spina bifida (SB), their parents, and in controls, to investigate which other enzymes of homocysteine metabolism might be defective. Because homozygosity for the 677C-->T mutation causes decreased plasma folate and increased red-cell folate (RCF) and plasma homocysteine levels, we excluded individuals homozygous for that mutation. The remaining SB patients and their parents still had lowered plasma folate and elevated total homocysteine levels, and a small subset had decreased vitamin B12 levels. Red-cell folate was the same in all groups, suggesting that dietary folate intake and its uptake was normal. Risk of SB was increased at the 25th percentile of plasma folate and at the 75th percentile of homocysteine values in SB patients and their parents, and at the 5th and 25th percentiles of vitamin B12 in mothers with SB-affected offspring. This underlines the functional importance of homocysteine remethylation to methionine. There was no correlation between vitamin B12 and homocysteine or RCF. In combination with the lowered plasma folate (80-90% 5-methyltetrahydrofolate), our data do not support a major involvement of methionine synthase in the aetiology of SB. Our data rather favour the involvement of genetic variation at loci coding for the formation of 5-methyltetrahydrofolate, such as MTHFR, methylenetetrahydrofolate dehydrogenase or serine hydroxymethyltransferase. PMID- 9327029 TI - Sequence analysis of the coding region of human methionine synthase: relevance to hyperhomocysteinaemia in neural-tube defects and vascular disease. AB - Elevated homocysteine (Hcy) levels are observed in two apparently unrelated diseases: neural-tube defects (NTD) and premature vascular disease. Defective human methionine synthase (MS) could result in elevated Hcy levels. We sequenced the coding region of MS in 8 hyperhomocysteinaemic patients (4 NTD patients and 4 patients with pregnancies complicated by spiral arterial disease, SAD). We identified only one mutation resulting in an amino acid substitution: an A-->G transition at bp 2756, converting an aspartic acid (D919) into a glycine (G). We screened genomic DNA for the presence of this mutation in 56 NTD patients, 69 mothers of children with NTD, 108 SAD patients and 364 controls. There was no increased prevalence of the GG and AG genotypes in NTD patients, their mothers or SAD patients. The D919G mutation does not seem to be a risk factor for NTD or vascular disease. We then examined the mean Hcy levels for each MS genotype. There was no correlation between GG- or AG-genotype and Hcy levels. The D919G mutation is thus a fairly prevalent, and probably benign polymorphism. This study, though limited, provides no evidence for a major involvement of MS in the aetiology of homocysteine-related diseases such as NTD or vascular disease. PMID- 9327030 TI - Plasma homocysteine, a risk factor for cardiovascular disease, is lowered by physiological doses of folic acid. AB - Elevated plasma homocysteine, an independent risk factor for cardiovascular disease (CVD) can be lowered by administration of pharmacological doses of folic acid. The effect of lower doses in apparently normal subjects is currently unknown but is highly relevant to the question of food fortification. Healthy male volunteers (n = 30) participated in a chronic intervention study (26 weeks). Folic acid supplements were administered daily at doses increasing from 100 micrograms (6 weeks), to 200 micrograms (6 weeks), to 400 micrograms (14 weeks). Fasting blood samples collected before, during and 10 weeks post intervention were analysed for plasma homocysteine, serum and red-cell folate levels. Results, expressed as tertiles of baseline plasma homocysteine concentration, showed significant (p < or = 0.001) homocysteine lowering in the top (10.90 +/- 0.83 mumol/l) and middle (9.11 +/- 0.49 mumol/l) tertiles only. In the low tertile, where the mean baseline homocysteine level was 7.07 +/- 0.84 mumol/l, no significant response was observed. Of the three folic acid doses, 200 micrograms appeared to be as effective as 400 micrograms, while 100 micrograms was clearly not optimal. There is thus a minimal level of plasma homocysteine below which folic acid has no further lowering effect, probably because an optimal folate status has been reached. A dose as low as 200 micrograms/day of folic acid is effective in lowering plasma homocysteine concentrations in apparently normal subjects. Any public health programme for lowering homocysteine levels, with the goal of diminishing CVD risk, should not be based on unnecessarily high doses of folic acid. PMID- 9327031 TI - Microvascular abnormalities in patients with vibration white finger. AB - In vivo nailfold capillary microscopy was performed on 10 men with vibration white finger (VWF) and 10 age-matched male controls. The observed nailfold capillaries required adaptation of Maricq's classifications and addition of new morphological scoring systems. These new classifications produced numerical scores for assessing capillary: dropout, tortuosity, elongation, visualization of subpapillary venular plexus, and the degree of disarrangement of nailfold capillary polarity. Application of these new scores showed, for the first time, a complex pattern of abnormal-nailfold capillaries in patients with VWF. Capillary dropout was evident in 7/10 patients, with an associated disarrangement in nailfold capillary polarity in five. All 10 controls had normal capillary morphology. Tortuosity of the capillary loops and elongation of their length was observed in 30% of patients. These significant morphological alterations seen in VWF suggest a local small-vessel vasculitis. PMID- 9327032 TI - Histiocytic necrotizing lymphadenitis, Kikuchi-Fujimoto's disease, associated with systemic lupus erythemotosus. AB - Histiocytic necrotizing lymphadenitis, Kikuchi-Fujimoto's Disease (KFD), is a condition rarely associated with systemic lupus erythematosus (SLE). The diagnosis of KFD can precede, postdate or coincide with the diagnosis of SLE. Lymphadenopathy is a common clinical presentation of KFD and SLE, and is histologically indistinguishable in both conditions. We describe two cases of KFD associated with SLE. The diagnosis of KFD in one case was made several years before the diagnosis of SLE, and the other was simultaneous. Both showed large lymphadenopathy, but neither fever nor neutropenia. Lymph-node biopsy showed necrosis, with proliferation of histiocytes and immunoblasts, paucity of neutrophils and absence of hemathoxilin bodies. Both patients responded favourably to steroid treatment. Patients with KFD should be assessed for SLE and have long-term follow-up checking for development of SLE. KFD should be ruled out in SLE flare-up accompanied by lymphadenopathy. PMID- 9327033 TI - Heparin therapy in the Chinese--lower doses are required. AB - Warfarin requirements are lower in the Chinese, but it is not known if this applies to heparin. We investigated the optimal dose for heparin therapy in Chinese patients, and to assess relationship between i.v. heparin dosage and anticoagulation efficacy. One hundred Chinese patients requiring intravenous heparin therapy were given an initial bolus followed by continuous intravenous infusion. The main outcome measures were: (i) Efficacy of anticoagulation assessed by blood coagulation studies (APTT) compared to heparin dosage, (ii) Determinants of dosage variation-age, gender, body weight, height, indication for heparin therapy and number of medications, other disease, and serum albumin level. It was found that the mean therapeutic infusion dose requirement of heparin was 848.7 +/- 274.7 units/h, 79% required a dose of 1000 units/h or less. Heparin dose correlated negatively with age (r = -0.40; p < 0.001) and positively with weight (r = 0.44 p < 0.001) and height (r = 0.49; p < 0.001). Chinese subjects require lower heparin doses (about 800 units/h) than usually recommended for Caucasians (usual dose 1000-1500 units/h). This can be partly explained by the lower body weight in Chinese patients. PMID- 9327034 TI - Biofeedback, relaxation training, and cognitive behaviour modification as treatments for lower functional gastrointestinal disorders. AB - Biofeedback, relaxation training, and cognitive behaviour modification are being increasingly proposed for the treatment of numerous functional disorders of the gastrointestinal tract. Among these, those related to the lower part of the gut seem to be more likely to benefit from this therapeutic approach. We examine and discuss the literature studies adopting such techniques. PMID- 9327035 TI - Vitamin B6 status, MTHFR and hyperhomocysteinaemia. PMID- 9327036 TI - The course of alcohol withdrawal in a general hospital. PMID- 9327038 TI - Bronchiectasis in Finland: trends in hospital treatment. AB - The incidence of bronchiectasis has probably declined in developed countries in recent years, but no reliable statistical data on this are available. The present paper describes the use made of hospital services by bronchiectatic patients in Finland. Data on a total of 12,539 treatment periods for bronchiectasis that had occurred between 1972 and 1992 were collected from the discharge register maintained by the National Research and Development Centre for Welfare and Health (diagnosis 518 in the International Classification of Diseases up to 1986, and 494 from 1987 onwards). The number of admissions, new occurrences of bronchiectasis and days in hospital were calculated by sex and age in relation to the total population at the end of each year. There were 143 and 87 admissions per million inhabitants in 1972 and 1992, respectively. The admissions, new occurrences and the days in hospital all decreased, at annual rates of 1.3, 4.2 and 5.7%, respectively. Thus, where the number of new occurrences was 50 per million persons in 1977, it was 27 per million in 1992. In summary, bronchiectasis-related hospital treatment declined markedly between 1972 and 1992. Trend is attributed to effective treatment of pulmonary infections and the reduction in tuberculosis. PMID- 9327037 TI - Phase 1 safety trial of Filgrastim (r-metHuG-CSF) in non-neutropenic patients with severe community-acquired pneumonia. AB - The objectives of the present study were to: (1) evaluate the safety of Filgrastim therapy in non-neutropenic patients with severe community-acquired pneumonia; (2) determine the absolute neutrophil count (ANC) response to various dosages of Filgrastim in non-neutropenic patients with active infection; and (3) describe the impact of therapy with Filgrastim in combination with antibiotics on selected pneumonia-related clinical parameters. The study design was an open label, dose-ranging, clinical trial, set in the General Clinical Research Unit of a large, public community hospital. The study population consisted of 30 patients who had presented to the Emergency Department with severe, community-acquired pneumonia. One of five dosages (75, 150, 300, 450 or 600 micrograms day-1) of Filgrastim (r-metHuG-CSF) was given subcutaneously daily for 10 days, until discharge or until the absolute neutrophil count > 75 x 10(9) l(-1), whichever was earlier. Vital signs, pulse oximetry, arterial blood gases, daily complete blood counts with differential, serum chemistries, coagulation profiles, electrocardiograms, chest radiographs, plasma G-CSF concentrations and duration of hospitalization were measured. There was no evidence of Filgrastim-related lung injury or evidence of extra-pulmonary toxicity. There was no apparent dose response effect of Filgrastim on pneumonia-related clinical variables. Dosages of Filgrastim between 150 and 600 micrograms day-1 had similar effects on increasing the ANC. Filgrastim appeared to be safe in non-neutropenic patients with severe, community-acquired pneumonia when given in dosages of 75-600 micrograms day-1 in combination with appropriate antibiotic therapy. Further study is needed to determine the effect of Filgrastim on morbidity, mortality and duration of symptoms in this patient population. PMID- 9327039 TI - Serum angiotensin converting enzyme does not correlate with radiographic stage at initial diagnosis of sarcoidosis. AB - Serum levels of angiotension converting enzyme (ACE) are elevated in many patients who suffer from sarcoidosis. Few studies have correlated ACE levels at diagnosis with the radiographic stage of the disease. The present authors reviewed the charts of all patients who had the diagnosis of sarcoidosis made between 1990 and 1995, and correlated ACE level at diagnosis with radiographic stage. Only patients with biopsy-proven sarcoid were included. One hundred and sixteen cases were identified, and complete data were available for 104 individuals. Serum ACE levels were increased in approximately 63.5% of the study population. The relationships between both stage and ACE level, and stage and percentage of individuals with elevated ACE levels within that stage were not statistically significant (P > 0.05). This large, retrospective study of patients with histologic evidence of sarcoidosis demonstrated no association between serum ACE level and radiographic stage. PMID- 9327040 TI - Artificial pneumothorax by the Veress cannula: efficacy and safety. AB - In 16 patients with pulmonary fibrosis, an artificial pneumothorax was introduced using the Veress cannula and the Saugman water manometer. Atmospheric air was introduced by fractionated insufflation to a total volume of 800 ml (median). The interpleural space was found on the first attempt, and in all cases, fractionated insufflation of atmospheric air was conducted while the intrapleural pressure was controlled with the water manometer. In one case, the procedure was stopped because of a rise in the pleural pressure after insufflation of only 50 ml air. This was undoubtedly caused by pleural adhesions not visible on chest X-ray. The main concern with air insufflation is air embolism but this was not observed clinically in any of the present cases. The patients in the present study all suffered from pulmonary fibrosis judged by clinical examination, chest X-ray and pulmonary function tests. Despite a diffusion capacity (DCO/VA) with a median value of 48% expected, the procedure was well tolerated. It has previously been shown that artificial pneumothorax preceding thoracoscopy is well tolerated due to hyperventilation, with an increase in respiratory frequency and a fall in arterial CO2 concentration (PaCO2), while pH and arterial O2 concentration (PO2) remain constant. This probably also explains the tolerance of the patients in this material. Insufflation of air as described here should be restricted to senior pulmonologists because it is an infrequent procedure. The present authors found the procedure to be uncomplicated and easy to perform with little discomfort to the patients. PMID- 9327041 TI - Thickness of the basement membrane of bronchial epithelial cells in lung diseases as determined by transbronchial biopsy. AB - The thickness of the basement membranes of bronchial epithelial cells varies under various pathological conditions. It has been reported that this membrane is thickened in patients with bronchial asthma. By light microscopy, this parameter was measured in biopsy specimens of bronchial mucosa obtained by fibre-optic bronchoscopy. These specimens were obtained from 171 patients who had undergone bronchial biopsy between 1984 and 1994. It was demonstrated that the thickness of the basement membrane of bronchial epithelial cells was weakly correlated with the patient's age, when thickness was examined in patients with lung cancer (r = 0.242, P = 0.0268). The basement membranes in patients with bronchial asthma (8.193 +/- 1.362 mu, mean +/- SEM) were significantly thicker than those without bronchial asthma (5.145 +/- 0.233 mu) (P = 0.0180, Mann-Whitney's U-test). In addition, it is noteworthy that the basement membranes in patients with diabetes mellitus (7.217 +/- 0.753 mu) were also significantly thicker than those without diabetes mellitus (4.968 +/- 0.235 mu) (P = 0.0038, Mann-Whitney's U-test). The background or underlying pathophysiology in such patients should be studied further, with attention directed towards the thickness of the bronchial basement membrane in bronchial biopsy specimens. PMID- 9327042 TI - Hospitalization of adults for asthma and inhaled corticosteroid use in an island population. AB - Inhaled corticosteroids have been shown to reduce morbidity and the need for hospitalization from asthma. Despite improvements in the therapy of asthma, epidemiologic data from several countries has shown that the hospital admission rates for asthma among adults at a population level are on the increase. The prevalence rate of hospital admission for asthma among Maltese adults aged 15-59 years was determined retrospectively from 1989 to 1993. Concurrent yearly total dispensal of inhaled corticosteroids for the whole population was also calculated. This study was undertaken amongst a well-defined island population served by a single medical facility offering emergency services, and a possible association between these two trends was investigated by means of logistic regression. The age-specific hospital admission rates for asthma decreased from 96.2 (95% CI: 109.7, 82.7) per 100,000 in 1989 to 38.1 (95% CI: 46.4, 29.8) per 100,000 in 1993. The prevalence rates of admission from asthma decreased from 67.6 (95% CI: 78.9, 56.3) per 100,000 in 1989 to 30.6 (95% CI: 38.0, 23.2) per 100,000 in 1993. The dispensal of inhaled beclomethasone dipropionate (BDP) increased from 0.99 defined daily dose (DDD) per 1000 population in 1989 to 3.28 DDD per 1000 in 1993. Logistic regression showed that increasing dispensal of inhaled BDP by 1 DDD per 1000 decreased the odds of an admission from asthma to 0.71 (95% CI: 0.65, 0.78) times their previous value. Similarly, the odds of an individual being hospitalized because of asthma decreased to 0.75 (95% CI: 0.67, 0.83) times their previous value. This study concludes that there was a progressive decrease in hospital admission rates for asthma in adults, and this trend correlates well with increasing use of inhaled corticosteroids at a community level. This must, however, be interpreted with care in light of the fact that increase in utilization of anti-inflammatory therapy probably also reflected improved general and widespread care for asthma. PMID- 9327043 TI - Asthma statistics in The Netherlands 1980-94. AB - Concern about the rising asthma mortality and morbidity in several countries in the 1980s, and the consequent development of international guidelines for diagnosis and management of asthma, gave reason to evaluate national mortality and hospital admission data from the Netherlands for asthma [International Classification of Diseases (ICD) 493] over the period 1980-94, as well as for chronic obstructive pulmonary disease (COPD) (ICD 490-2, 496) and acute bronchi(oli)tis (ICD 466), according to age (0-4, 5-34, 35-64 and > or = 65 years). Rates per million population per year were calculated and time trend analyses were performed. Hospital admissions for asthma showed a decrease in all age groups except in age group 0-4 years. In this age group, an increase was found which continued in the 1990s. Hospital admissions also increased for COPD and acute bronchi(oli)tis in the age group 0-4 years. These increases, however, had no impact on the respiratory mortality, which remained stable and even fell for acute bronchi(oli)tis. Asthma mortality showed a large decline in the 1990s in age group > or = 65 years in both sexes, and also fell, to a lesser extent, in age group 35-64 years. In both age groups, rising COPD trends were found in hospital admissions and mortality, except in males aged 35-64 years, in whom trends fell in the last decade. In age group 5-34 years, asthma mortality declined over the whole study period, whilst the other respiratory trends were stable. It is concluded that asthma statistics in the Netherlands indicate favourable development, except for the age group 0-4 years. In this age group, morbidity from asthma and from other reactive airway disorders is still of great concern. PMID- 9327044 TI - Phase II study of weekly gemcitabine in advanced non-small cell lung cancer. AB - New active agents are needed to develop effective systemic therapy against Stage IIIB-IV non-small cell lung cancer (NSCLC). The aim of the present study was to assess the efficacy and toxicity of gemcitabine, a novel nucleoside analogue with significant preclinical activity, as a single-agent therapy. Forty-three patients with previously untreated Stage IIIB-IV NSCLC were included. Gemcitabine was administered intravenously over 30 min on Days 1, 8 and 15 of each 28-day cycle at a dose of 1250 mg m-2. Thirty-seven patients were evaluable for response. There were seven partial responses giving an overall response rate of 19% (95% confidence interval 8-35%). Median duration of response was 6 months. One-year survival and median survival for all patients were 33% and 8 months, respectively. Toxicity of the treatment was mild. World Health Organization (WHO) Grade 3-4 leukopenia was detected in 11% of the patients. Mild (WHO Grade 1-2) nausea was the most frequent subjective side-effect with a rate of 82%. Mild rash and peripheral oedema were typical side-effects of gemcitabine with rates of 19 and 9%, respectively. In conclusion, single-agent gemcitabine is an active and well-tolerated treatment for Stage IIIB-IV NSCLC patients. PMID- 9327045 TI - Contemporary use of antibiotics in 1089 adults presenting with acute lower respiratory tract illness in general practice in the U.K.: implications for developing management guidelines. AB - Respiratory symptoms are the most common cause of general practitioner (GP) consultation, and hospital-based specialists are often called on to provide management guidelines, particularly in the area of antibiotic prescribing. The present authors have assessed factors associated with antibiotic use by 115 GPs when managing 1089 adults with an acute lower respiratory tract illness, including cough. They prescribed antibiotics to three-quarters of patients, but felt antibiotics to be definitely indicated in less than one-third of these cases and not needed in one-fifth. Univariate analysis revealed that antibiotics were prescribed more frequently by older GPs for older patients in the presence of underlying disease, discoloured sputum, shortness of breath, wheeze, fever, signs on chest examination, and 'other factors'. Multivariate logistic regression confirmed an independent effect for all these findings except for the presence of underlying disease, shortness of breath and wheeze. 'Other factors' included patient 'pressure' and social factors, and GP work pressure or prior experience with the patient. These factors were an important influence on prescribing, especially if the GP felt an antibiotic was not indicated. Amoxycillin was the first choice (58% of total) except where the patient had recently received antibiotics for the same illness. Broader spectrum antibiotics were used more commonly in patients with chronic lung disease, discoloured sputum, chest signs on examination and where the GP felt antibiotics were indicated. However, these antibiotics were also prescribed to 14% of previously well patients. General practitioners used a wide variety of terms to describe the illness with little consistency or structure. The decision concerning the use and choice of antibiotics and the confidence with which the GP makes that decision is a complex interaction between patient, doctor and disease, being affected not only by clinical features but also by the social and psychological elements of the presenting problem. Such issues need to be appreciated by hospital specialists when called on to advise on developing relevant guidelines for primary care. PMID- 9327047 TI - A case of acute deterioration in asthma symptoms induced by isoniazid prophylaxis. AB - The present case report describes a 10-year-old boy with clinical history of steroid-dependent asthma who developed severe exacerbation of his respiratory symptoms upon isoniazid administration. Subsequent control of his asthma symptoms was re-established and maintained only after isoniazid withdrawal. This case is the first to emphasize the dangers of isoniazid administration in patients who have asthma. PMID- 9327046 TI - Invasive pulmonary aspergillosis with cerebromeningeal involvement after short term intravenous corticosteroid therapy in a patient with asthma. AB - The present case report describes a case of invasive pulmonary aspergillosis with cerebromeningeal invasion in an asthmatic non-immunocompromised patient. This fatal complication occurred despite early anti-fungal antibiotherapy after a 2 week course of intravenous corticosteroid therapy for treatment of an exacerbated asthma. Diagnostic and therapeutic procedures are discussed. PMID- 9327048 TI - Contrast and spatial-frequency requirements for emmetropization in chicks. AB - This study examined the contrast and spatial-frequency requirements for emmetropization in chicks. Chicks were form deprived from hatching either constantly or had this treatment interrupted with 20 min of "visual stimulation" each day. Visual stimulation comprised exposure to either a normal cage environment (i.e., normal vision) or environments that were restricted in either their spatial contrast or spatial-frequency composition. Constant form deprivation resulted in high myopia (e.g. -11.8 D after 5 days), with refractive changes being much smaller in chicks allowed 20 min of normal vision each day (e.g. -3.4D). The restricted contrast environments (contrast range: 9-78%) were generally only slightly less effective than the normal cage environment in preventing form-deprivation myopia. However, in the case of restricted spatial frequency environments, both the intermediate (0.86 cycles deg-1) and mixed spatial-frequency environments significantly reduced the form deprivation response, while both the high (4.3 cycles deg-1) and low spatial-frequency (0.086 cycles deg-1) stimuli, as well as the composites of these, were less effective in preventing form-deprivation myopia. This spatial-frequency dependence did not vary when, instead of white light, monochromatic illumination was used to eliminate chromatic aberration, although all groups showed more myopia under this condition. It is assumed that the observed inhibitory effects on form-deprivation myopia reflect the adequacy of the visual information presented during the period of visual stimulation for emmetropization in chicks. In this context, the data imply a mid-spatial-frequency tuning in the current study and a low contrast threshold which was not reached for this emmetropization process. Finally, the data hint that chromatic aberration may have some role as a cue to defocus in emmetropization. PMID- 9327049 TI - Properties of glutamate-gated ion channels in horizontal cells of the perch retina. AB - The effect of two different concentrations of L-glutamate and kainate on the gating kinetics of amino acid-sensitive non-NMDA channels were studied in cultured teleost retinal horizontal cells by single-channel recording and by noise analysis of whole-cell currents. When the glutamate agonist kainate was applied clearly parabolic mean-variance relations of whole-cell membrane currents (up to 3000 pA) indicated that this agonist was acting on one type of channels with a conductance of 5-10 pS. The cells were less sensitive when L-glutamate was used as the agonist and in most cases whole-cell currents amounted to less than 200 pA. The mean-variance relation of glutamate induced currents was complex, indicating that more than one type of channel opening could be involved. Power spectra of whole-cell currents were fitted with two Lorentzians with time constants of approx. 1 and 5-20 msec. Effects on amplitudes and time constants of agonist concentrations are demonstrated. Two categories of unitary events with mean open times of approx. 1 and 7 msec and conductances of approx. 7 and 12 pS, respectively, were obtained in single-channel recordings from cell-attached patches at different concentrations of glutamate in the pipette. PMID- 9327051 TI - Sharpness overconstancy in peripheral vision. AB - Although much has been learned about the spatial sampling and filtering properties of peripheral vision, little attention has been paid to the remarkably clear appearance of the peripheral visual field. To study the apparent sharpness of stimuli presented in the periphery, we presented Gaussian blurred horizontal edges at 8.3, 16.6, 24, 32, and 40 deg eccentricity. Observers adjusted the sharpness of a similar edge, viewed foveally, to match the appearance of the peripheral stimulus. All observers matched blurred peripheral stimuli with sharper foveal stimuli. We have called this effect "sharpness overconstancy". For field sizes of 4 deg, there was greater overconstancy at larger eccentricities. Scaling the field size of the peripheral stimuli by a cortical magnification factor produced sharpness overconstancy which was independent of eccentricity. In both cases, there was a slight sharpness underconstancy for peripherally presented edges blurred only slightly. We consider various explanations of peripheral sharpness overconstancy. PMID- 9327050 TI - Reticalmin: a novel calcium/calmodulin-dependent protein kinase IV-like protein in rat retina. AB - Western blot analysis of 100,000 g supernatant of rat retina using a polyclonal anti-Ca2+/ calmodulin-dependent protein kinase IV (CaM-kinase IV) antibody revealed an immunoreactive mass of 35 kDa, termed reticalmin. Lower amount of a isoform of CaM-kinase IV was also expressed in rat retina. Reticalmin did not react with anti-CaM-kinase IV C-terminal peptide antibody which recognized alpha and beta isoforms of CaM-kinase IV and calspermin. Immunohistochemically reticalmin was shown to be localized mainly in the outer segment of photo receptor cells, and in dendrites of inner plexiform layers and may be in nuclei of ganglion cells and some inner nuclear layer cells. PMID- 9327052 TI - Direction discrimination of cyclopean (stereoscopic) and luminance motion. AB - This study compared direction discrimination of cyclopean (stereoscopic) and luminance motion involving stimuli equated for effective strength. The stimuli were random-walk cinematogram (RWC) displays whose signal and noise discs were created from binocular disparity differences embedded in a dynamic random-dot stereogram or from luminance differences. Experiment 1 measured global motion detection thresholds for cyclopean and luminance stimuli by manipulating the proportion of signal to noise discs. Detection thresholds for cyclopean motion were about 25% whereas detection thresholds for luminance motion were 5%, thus five times more cyclopean motion events than luminance events were necessary to elicit threshold responding. Experiment 2 measured thresholds for discriminating the direction of cyclopean and luminance motion under conditions of equal stimulus strength by presenting the motion displays at equal multiples of detection threshold. Direction discrimination thresholds (ranging from about 5-30 deg, depending upon conditions) were similar for cyclopean and luminance motion, thus the precision with which the pooling of local motion events in one direction can be discriminated from the pooling of events in a slightly different direction is the same for cyclopean and luminance stimuli. The finding that cyclopean motion information is pooled is consistent with the idea that the direction of cyclopean motion is coded in the responses of a population of directionally selective mechanisms. PMID- 9327053 TI - Is stereopsis effective in breaking camouflage for moving targets? AB - It has been suggested that breaking camouflage is one of the major functions of stereopsis (Julesz, 1971). In this study, we found that stereopsis is less effective in breaking camouflage for moving targets than for static ones. Observers were asked to detect a single dot moving on a straight trajectory amidst identical noise dots in random motion. In the three-dimensional (3D) condition, the noise dots filled a cylindrical volume 5.7 cm in height and diameter; the trajectory signal dot moved on an oblique 3D trajectory through the center of the cylinder. In the two-dimensional (2D) control condition, observers viewed one half-image of the 3D cylinder binocularly. Surprisingly, trajectory detection in the 3D condition was only slightly better than in the 2D condition. Stereoscopic tuning for motion detection was also measured with a novel target configuration in which the random motion noise was presented in two depth planes that straddled the fixation plane where the trajectory target was presented. As the disparity between the noise planes and the fixation plane was increased, trajectory detection improved, reaching a peak between 6 and 12 arcmin, and then declining to the 2D level at larger disparities, where the noise became diplopic. Similar tuning measurements were made for detecting a static pattern, a string of five aligned dots presented in the fixation plane between two planes of static noise dots. Adding disparity to the noise planes produced a far greater improvement in static detection than in motion detection, for a comparable range of disparities (1.5-12 arcmin). We speculate that the temporal characteristics of the stereo system are not well suited for responding to moving targets, with the result that stereo does not greatly enhance motion detection in noise. PMID- 9327054 TI - The development of chromatic and achromatic contrast sensitivity in infancy as tested with the sweep VEP. AB - Swept-contrast visual evoked potential (VEP) techniques were used to measure the development of contrast sensitivity functions (CSFs) for achromatic and red/green isoluminant chromatic gratings. Subjects were infants of 8, 14, 20 and 32 weeks of age, and adults. Stimuli were 20 deg, 0.3-4 cyc/deg sinusoidal gratings, counterphased at 6 Hz and modulated through white. Achromatic and chromatic CSFs for all ages could be fit simultaneously with a double exponential equation of a common, lowpass shape. Both achromatic and chromatic CSFs exhibited developmental shifts in both sensitivity and spatial scale. From 8 weeks to adulthood, sensitivity increased by 0.64 log units for achromatic gratings and by 0.91 log units for chromatic gratings, yielding an 0.27 log unit larger sensitivity change for chromatic than for achromatic stimuli. Spatial scale shifts were closely similar across achromatic and chromatic CSFs, and were consistent with the factor of about four predicted on the basis of changes in foveal receptor packing density and eye size. The question of uniform vs differential loss of sensitivity for chromatic vs achromatic stimuli at fixed spatial frequencies is discussed. PMID- 9327055 TI - Isoluminance and chromatic motion perception throughout the visual field. AB - Isoluminance and chromatic motion perception for red/green gratings were measured throughout an 80 deg visual field. Generally, the red/green isoluminance values changed with increasing eccentricity, i.e., observers increased the red luminance contrast for a fixed green luminance contrast. Enlarging the target size (to compensate for the cone density changes with eccentricity) and decreasing the spatial frequency (to compensate for receptive field property changes with eccentricity) did not change the isoluminance values within the central 20 deg, but the isoluminance ratios decreased beyond 20 deg. Our manipulations did not entirely compensate for a given eccentricity, which implies the need for a post receptoral scaling function for the perception of drifting chromatic stimuli. Further, the results for isoluminance show heterogeneity between the visual field meridians where the red to green luminance ratio tends to be greater in the superior visual field. In our present conditions, chromatic motion was always perceived (up to 40 deg of eccentricity), but sensitivity generally decreased with increasing eccentricity. The inferior visual field was found to be the most sensitive to chromatic motion. We propose that the lower visual field and not the superior visual field is specialized for colour motion information. PMID- 9327056 TI - Visual search of expansion and contraction. AB - The perception of expansion/contraction in human subjects was examined with a visual search paradigm. When searching for a target defined by two-dimensional expansion among distractors defined by two-dimensional contraction, the time needed to find the target did not vary as the number of distractors was increased. However, for a target defined by two-dimensional contraction among distractors defined by two-dimensional expansion, the search time increased as a function of the number of distractors in the display. A similar search asymmetry remained between one-dimensional expansion and one-dimensional contraction, even though one-dimensional expansion was searched in a serial manner. This asymmetry between expansion and contraction reflects a basic characteristic of higher-order motion information processing. PMID- 9327058 TI - Edge detection by landing honeybees: behavioural analysis and model simulations of the underlying mechanism. AB - The mechanism of edge detection in the honeybee was investigated by examining the effects of combining different kinds of visual cues that define an edge. Free flying bees were trained to land at three different types of edges which were defined by texture and relative motion cues either in isolation or in combination with each other. Bees are able to detect and land at the three types of edges, but do so with different frequencies. In contrast to the naive expectation that edges jointly defined by two cues can be detected better than those defined by a single cue in isolation, the combination of the cues does not increase and may even decrease the detectability of an edge. When bees land at an edge the orientation of their body axis is strongly affected by the visual cues defining this edge. Model simulations were performed to test whether the experimental findings can be explained on the basis of a single edge detection mechanism sensitive to both types of visual cues. In the model, the information from both types of cues is sensed by two fields of movement detectors that receive their input signals from two adjacent patches in the visual field. The output of all detectors subserving either patch is pooled by integrating cells. The signals of the two integrating cells subserving the two adjacent patches are compared at a subtraction stage. The resulting signal is then rectified and forms the output signal of the model. The model simulations closely resemble the experimental results, thus providing evidence that edge detection by the bee could be mediated by a single mechanism. PMID- 9327057 TI - Effects of dot density, patch size and contrast on the upper spatial limit for direction discrimination in random-dot kinematograms. AB - Two-frame random-dot kinematograms (RDKs) of different dot density, area and contrast were used to study the spatial properties of the human visual motion system. It was found that the maximum spatial displacement at which observers could reliably discriminate the direction of motion (dmax) increased gradually by a factor of up to 6.4 as dot density was decreased from 50 to 0.025% for high Michelson contrast (0.997) stimuli. As stimulus area was reduced from 645 deg2, this trend gradually disappeared so that by a stimulus area of 2.56 deg2, there was no effect of density upon dmax. A further experiment investigated the effects of reducing Michelson contrast from 0.77 to 0.2 on dmax over this same range of dot densities. It was found that at the highest densities, dmax declined as contrast was reduced. Furthermore, for contrasts at and below 0.4, dmax was invariant of density over the range 50-5%. These results can be accounted for by the fact that both reducing contrast, while keeping density fixed, and reducing density, while maintaining a fixed high contrast, reduce the stimulus mean luminance. For all contrasts, decreasing density below 5% led to an increase in dmax. However, the rate of this increase was slower for the lower contrast stimuli. A two-stage model based on bandpass filtering followed by an informationally limited motion detection stage is proposed and shown to provide a good account of these data. PMID- 9327059 TI - Saccadic reaction times: a statistical analysis of multimodal distributions. AB - The distributions of saccadic reaction times (SRT) often deviate from unimodal normal distributions. An excess-mass procedure was used to detect peaks in 963 data sets containing 90,927 reaction times from 170 subjects. About 55% showed one, 30% two, 12% three and 3% four peaks. According to their clustering along the reaction time scale the modes could be classified into express (90-120 msec), fast regular (135-170 msec) and slow regular (200-220 msec) modes. Among the unimodal distributions 29% had peaks in the range of the express mode and 46% had peaks in the range of the fast regular mode. Therefore, 87% of the data sets support the notion of saccadic reaction time distributions being the superposition of three modes. All experimental distributions were fitted by as many gamma distributions as determined by the excess-mass test. The significance of the multimodality for saccade generation processes is discussed. PMID- 9327060 TI - The role of feedback in learning a vernier discrimination task. AB - We compare improvement through training in vernier acuity under different feedback conditions in order to clarify the role of feedback during learning of a perceptual task and to test different (neural network) models of perceptual learning. Improvement of performance is measured in 49 observers under feedback, no feedback, uncorrelated feedback, partial feedback, and block feedback conditions. Correct feedback conditions yield a larger improvement of performance than manipulated and no feedback conditions. Providing feedback that is uncorrelated to the observers' responses prevents learning, while the effect of block feedback does not differ significantly from complete feedback. Our results cannot be explained by learning rules that depend exclusively on an external teacher or by models that propose learning in an exposure-dependent way with unsupervised learning rules but without top-down influences. PMID- 9327061 TI - Computing stereo channels from masking data. AB - The detection of stereoscopic depth in random-dot patterns that have been spatially band-pass filtered is adversely affected by the addition of noise at spatial frequencies in the neighbourhood of the frequencies present in the stereogram. This elevation of threshold is generally termed masking and recent data have been interpreted as evidence for a pair of spatial-frequency-tuned stereo "channels" whose peak spatial frequencies are either at 3 and 5 c/deg or 2.5 and 7 c/deg. This interpretation was re-examined. In particular, we have studied how the characteristics of masking interactions might be affected by taking account of the presence of an initial modulation transfer function (including the optical m.t.f. of the eye) that precedes the stage at which signal and mask interact. Using this approach, we reach the conclusion that the peak of the internal masking function for stereo detection coincides with the signal spatial frequency over the whole range tested (1.7-11.6 c/deg). We conclude that the recent data of Yang and Blake [(1991). Vision Research, 31, 1177-1189] are consistent with a multiple channel model in much the form proposed by Julesz and Miller [(1975). Perception, 4, 125-143]. The analysis presented in this paper has general implications for the interpretation of masking studies in spatial contrast vision. PMID- 9327062 TI - What spatial frequency do we use to detect the orientation of a Landolt C? AB - We calculated the two-dimensional Fourier spectrum of a Landolt C. For Landolt Cs of orthogonal orientation, the main differences in the amplitude spectrum were found at a low frequency such that 1.3 periods were equal to the size fo the Landolt C, rather than at the high frequency corresponding to the size of the gap, i.e., such that 2.5 periods were equal to the size of the Landolt C. We compared visual acuity assessments obtained with Landolt C optotypes and the cut off of the contrast sensitivity function measured with sinusoidal gratings. We found that the frequency corresponding to the size of the gap is twice the latter frequency. We suggest that, in fact, this lower frequency can be used to determine the position of the gap. PMID- 9327063 TI - Cone electroretinogram to chromatic stimuli in myopic eyes. AB - The cone electroretinograms (ERGs) to different chromatic stimuli were recorded in myopic subjects. Ganzfeld color flashes under bright white background illumination were used to elicit short-wavelength-sensitive (S-), and mixed long (L-) and middle-(M-) wavelength-sensitive cone ERGs. Nineteen subjects with mild myopia (between -3.0 and -6.0 D), 12 subjects with high myopia (greater than 6.25 D) but without chorioretinal atrophy or posterior staphyloma, and 22 age matched normal controls were compared. The S-cone and L,M-cone b-wave amplitudes decreased progressively with increasing myopia. The amplitudes of the S-cone and the L,M-cone b-wave were significantly lower in highly myopic subjects, as compared with controls. The implicit times of both b-waves were normal in most myopic subjects. The S-cone and the L,M-cone ERGs were almost equally affected in myopic eyes. The selective reduction of the S-cone pathway in high myopia, reported previously, may not originate from the other retina. PMID- 9327064 TI - Separate magnocellular and parvocellular contributions from temporal analysis of the multifocal VEP. AB - Temporal analysis of the multifocal cortical visual evoked potential (VEP) was studied using pseudo-random (m-sequence) achromatic stimulation. The effects of variation of luminance contrast on the first-order response were complex. At low to mid contrasts (< 60%), a wave doublet (P100-N115) predominated. A second wave complex (N100-P120-N160) dominated at high contrasts. The second-order responses, however, showed an extremely simple variation with luminance contrast. Intrinsic differences in the adaptation time of the generators of these two components caused a distinct separation in the slices of the second-order response. A rapidly adapting nonlinearity saturating at low contrasts was only observable when measuring the responses from two consecutive flashes. Its latency coincided with the contrast saturating first-order response component. By comparison, the nonlinearity derived from the responses to the stimuli with longer interstimulus intervals (second and third slices) yielded a much more linear contrast response function with lower contrast gain and latencies, which clearly corresponded to the longer latency component of the first-order response. Thus, the second-order responses show a first slice which is predominantly driven by neural elements that have a latency and contrast function that mimic those of the magnocellular neurons of the primate LGN and a second slice which is dominated by a generator whose properties resemble primate parvocellular function. This division into magno and parvocellular contribution to the VEP is based on function (interaction time) as distinct from other currently available analyses, with potential for neural analysis of visual disease. PMID- 9327065 TI - Unintentional fatal paraquat poisonings among agricultural workers in Costa Rica: report of 15 cases. AB - This study analyzes the exposure circumstances of 15 fatal occupational paraquat poisonings. To evaluate the potential danger of dermal absorption and the amount needed to produce a fatal outcome in the event of oral intake, we reviewed the medical records and autopsy protocols and interviewed relatives. Five fatalities were due to ingestion of a mouthful of paraquat concentrate, and five to intake of a smaller amount; three cases were associated with dermal exposure, and in two, there was no evidence of either oral or dermal exposure. Several cases concerned diluted paraquat spray. The clinical and pathomorphological findings, including a "blinded" evaluation of lung slides, were consistent with paraquat poisoning in all cases. Difficulties in establishing the diagnosis and recognizing the exposure were identified, as well as classification of unintentional poisonings as suicides at autopsy. The findings suggest that paraquat may cause fatal poisonings by ingestion of small amounts, by dermal absorption of diluted paraquat, and possibly by inhalation. More conclusive studies are warranted. PMID- 9327066 TI - Mortality from a Chinese asbestos plant: overall cancer mortality. AB - The cancer mortality in a Chinese asbestos plant using only chrysotile was studied. Opened in the 1950s, all workers with at least 1 year of employment by 1972 were followed through 1994. Most workers were female. Excess cancer mortality, compared to local city data, was found for lung cancer, including many cases among nonsmoking women, and stomach cancer. Plant asbestos levels have been high in the past but have consistently come down over the decades. PMID- 9327067 TI - Pleural plaques as risk indicators for malignant pleural mesothelioma: a necropsy based study. AB - Pleural plaque is recognized as a reliable marker of previous exposure to asbestos. However, it is controversial whether pleural plaque is a risk indicator for asbestos-related malignancies. In the present study, the thoracic cavities were examined for pleural plaques in 3,005 necropsies performed at the Monfalcone Hospital in people aged 15 years or older. Plaques were classified into three classes: 1, small (plaques measuring 1-4 cm in major diameter); 3, large (plaques involving a major part of a hemithorax); and 2, moderate (intermediate conditions). The prevalences of pleural plaques were 70.9% among men, and 24.0% among women. The prevalences of plaques (total plaques, various classes) among subjects with pleural mesothelioma were compared with those observed in the remaining cases. The series included 92 subjects with malignant pleural mesothelioma (82 men and 10 women). Mesothelioma cases showed higher prevalences of total plaques as well as higher prevalences of classes 1, 2, and 3, when compared with controls. These differences reached the statistical significance for total plaques, and classes 2, 3. The present data are consistent with the idea that pleural plaque is a risk indicator for pleural mesothelioma. PMID- 9327068 TI - Respiratory health of automobile workers exposed to metal-working fluid aerosols: respiratory symptoms. AB - A total of 1,811 automobile workers at three General Motors facilities were evaluated by questionnaire for possible respiratory effects resulting from airborne exposures to metal-working fluids (MWF): 1,042 currently worked as machinists and were exposed to one of three types of MWF aerosols (straight mineral oils, soluble oil emulsions, or water-based synthetic fluids that contained no oils); 769 assembly workers, without direct exposure, served as an internal reference group (of these, 239 had never worked as machinists). Symptoms of usual cough, usual phlegm, wheezing, chest tightness, and breathlessness, as well as physician-diagnosed asthma, and chronic bronchitis were the primary outcomes examined. Machinists as a whole had higher prevalence of cough, phlegm, wheezing, and breathlessness than that of assembly workers. Adjusting for confounding, phlegm and wheeze were associated with increasing levels of current exposure to straight oils; cough, phlegm, wheeze, chest tightness, and chronic bronchitis were associated with increasing levels of current exposure to synthetics. In models that included both past and current exposure, only current exposures to straight and synthetic fluids were associated with current symptoms. PMID- 9327069 TI - Loss of lung function among sheet metal workers: ten-year study. AB - One hundred and twenty-two sheet metal workers in New England were examined over a 10-year interval for loss of pulmonary function and the development of asbestosis or asbestos-related pleural fibrosis. Regression models using the generalized estimating equation (GEE) approach were created to investigate the relationship between exposure and pulmonary function after adjusting for smoking status, age, height, and asbestos-related x-ray changes. A history of shipyard work was a significant contributor to the loss of forced vital capacity (FVC). Among smokers, loss in forced expiratory volume at 1 sec (FEV1) also had a significant relationship to prior shipyard work. There was a borderline significant relationship between percentage predicted FEV1 and cumulative years of asbestos exposure in smokers, as well as years-since-initial-exposure in never smokers. This study supports previous findings of obstructive airway changes in asbestos-exposed workers and identifies shipboard work as an important predictor of loss in pulmonary function even years after shipyard exposure to asbestos has ceased. PMID- 9327070 TI - Cancer morbidity in workers at aluminum foundries and secondary aluminum smelters. AB - In a Swedish cohort of workers (n = 6,454) from seven aluminum foundries and three secondary aluminum (scrap) smelters there was no overall excess risk of cancer among male or female workers less than 85 years of age (males: 325 observed cases, standardized incidence ratio (SIR) 1.02, 95% confidence interval (CI) 0.91-1.13; females: 22 cases, SIR = 0.95, 95% CI = 0.60-1.44). In male workers, however, significantly elevated risk estimates were observed for cancer of the lung (51 cases; SIR = 1.49, 95% CI = 1.11-1.96), anorectal cancer (33 cases; SIR 2.13, 95% CI = 1.47-2.99), and sinonasal cancer (4 cases; SIR = 4.70, 95% CI = 1.28-12.01). There was no increase of urinary bladder or liver cancer. Lung cancer risks were highest in workers with a short duration of employment (< 5 years) suggesting determinants of risk related to socioeconomic factors rather than the occupational environment under study, but there were also indications of a lung cancer hazard from sand casting of aluminum for 10 years or more (SIR = 2.10, 95% CI = 1.01-3.87). The increase in anorectal cancer could not be etiologically related to occupational determinants of risk. Sand casting of aluminum aside, the cancer risk in secondary aluminum smelting seems to be lower than in primary aluminum smelting and in iron and steel founding, respectively. PMID- 9327071 TI - Serum liver function profiles in coking workers. AB - Coking workers are regularly exposed to coke oven emissions (COE), which consist mainly of polycyclic aromatic hydrocarbons and volatile organic compounds. In a previous cross-sectional study, we found that coking and by-product workers with heavy exposure to COE in the older of two coke operation areas in Taiwan had higher serum activities of hepatic aminotransferase than the controls. In this study, we further examine the relationship of exposure to COE with liver function profiles in coking workers. Liver function profiles included serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), and total bilirubin (BIL). The exposed group included 88 workers working 3 months or more in the older coke oven plant. Fifty-nine referents, not visiting the coke operation areas in the last 3 months, came from the administrative area in the same company. Each participant wore a personal monitor that was used to measure benzene soluble fraction (BSF) of total particulates, as a surrogate of COE, for 3 consecutive days between August 1995 and February 1996. Serum liver function profiles, hepatitis B surface antigens, and anti-hepatitis C antibodies were examined in the morning following the exposure measurements. Exposure levels were categorized by exposure situations (high, medium, low) among coking workers. The high exposure group (n = 23) worked topside of the oven. The medium exposure group (n = 44) worked at the sideoven for more than 4 hr/day, whereas the low exposure group (n = 21) worked at the sideoven for less than 4 hr/day and mostly remained in the control rooms. The low exposure group was used as an internal comparison group. The median BSF concentrations for various exposure situations were as follows, high exposure group: 372 micrograms/m3, medium exposure group: 61 micrograms/m3, low exposure group: 49 micrograms/m3, and referents: 10 micrograms/m3. The coking workers (n = 88) did not significantly differ from the referents (n = 59) in any of the liver function profiles. Excluding the referents, workers in the high exposure group had a mean AST level that was 31% higher (95% confidence interval (CI) = 9-57%) and a mean ALT level that was 46% higher (95% CI = 7-98%) than those in the low exposure group after adjusting for appropriate confounders in multivariate models. The prevalence of an abnormal hepatocellular pattern (AST > 37 IU/L or ALT > 39 IU/L) was more common in the high exposure group than in the low exposure group (adjusted odds ratio = 4.4; 95% CI = 0.9-22.6). However, these associations were not found in GGT, ALP, or BIL. After controlling for the possible effects of nonoccupational factors on serum activity of AST and ALT, we conclude that increased AST and ALT levels among topside coking workers may be caused by heavy inhalation exposure to COE. Additionally, the adverse hepatic effect seems to be caused by a mixture of hazards, rather than a unique identifiable chemical. PMID- 9327072 TI - Long-term use of organophosphates and neuropsychological performance. AB - This study evaluated neuropsychological effects due to chronic organophosphate use among farmers with no history of acute poisoning. Fifty-seven male tree fruit farmers (exposed) were compared with 42 age-matched male cranberry/blueberry growers and hardware store owners (unexposed). Univariate analyses of covariance (reading test as covariate) comparing exposed and unexposed subjects revealed significantly slower reaction time. No other significant differences were noted on tests of concentration, visuomotor skills, memory, expressive language, or mood. Based on an exposure metric derived from detailed exposure histories, farmers were divided into high exposure (n = 40) and low exposure (n = 59) groups, and their neuropsychological performance was compared. Analysis of covariance with age and reading test score as covariates revealed that the high exposure group had significantly slower reaction time, dominant hand. Long-term use of organophosphates without evidence of an acute poisoning episode appears to produce, at most, subtle changes in neuropsychological performance. PMID- 9327073 TI - Coxarthrosis and farm work: a case-referent study. AB - The purpose of this case-referent study was to analyze the association between coxarthrosis and occupation. The study was performed in a Swedish agricultural county and comprised 269 cases of radiologically verified arthrosis of osteoarthritis (< 3 mm joint space) that were compared to 538 randomly selected controls in the same region, matched for age, sex, and place of residence. Farmers and agricultural workers showed an increased risk of coxarthrosis and the observed risk increased with increasing number of years of farming. Tractor driving and milking were associated with coxarthrosis, whereas no association with other types of machine work could be demonstrated. An association between coxarthrosis and heavy physical work before the age of 16 years was also observed. The results give only limited information on the external causes of coxarthrosis, however. More detailed studies of groups of individuals with coxarthrosis in order to obtain more information about contributing and underlying factors would therefore be valuable. PMID- 9327074 TI - Hospital costs associated with agricultural machinery injuries in Ontario. AB - To assist those responsible for agricultural safety, we: (1) piloted an approach to costing hospitalized farm injuries; and, (2) described ambulance and inpatient costs associated with these injuries in Ontario. Hospital discharge records (hospital separations) for farm machinery injuries in Ontario (n = 1,610) were identified by ICD9-CM E-codes for 1985-1993. Ambulance costs were estimated by the Ontario Ministry of Health. For each case, the hospital costs were calculated by multiplying the case-specific resource intensity weight by the average inpatient cost per weighted case. The costs (1993 Canadian dollars) ranged from $768 to $62,643 and totaled $6.9 million over the study period. Males accounted for 89.8% of the total costs. Tractor injuries accounted for a large proportion of costs (34.3%). The median costs per case varied by type of machinery, ranging from $2,043 for ploughs/disks to $3,366 for augers. Entanglement injuries were responsible for the largest proportion of costs (40.7%), while tractor rollovers accounted for the highest median cost ($3,065). Although these figures represent a fraction of the total costs associated with farm injuries, the results provide one basis from which to justify and target preventive initiatives. This approach to costing may also be widely applicable to other health issues. PMID- 9327075 TI - Meta-analyses of multiple myeloma and farming. AB - A series of meta-analyses of peer-reviewed studies of multiple myeloma (MM) and farming were performed, using 32 studies published between 1981 and 1996. Prior to the meta-analyses, all studies were reviewed and evaluated for heterogeneity and publication bias. A random-effects meta-analysis including all of the studies yielded an estimator of relative risk equal to 1.23, with a 95% confidence interval (95% CI) of 1.14, 1.32. The estimator of relative risk obtained from a meta-analysis restricted to female farmers was 1.23 (95% CI = 1.17, 1.29). A third meta-analysis restricted to studies of farmers residing in the central United States resulted in an estimator of relative risk equal to 1.38 (95% CI = 1.27, 1.51). These findings were not influenced by either a publication bias or a specific study design. The consistent significant positive findings suggest that there is an association between MM and farming. Exposures commonly experienced by farmers and that might contribute to the occurrence of MM include infectious microorganisms, solvents and pesticides. PMID- 9327076 TI - Assessment of the relationship between isocyanate exposure levels and occupational asthma. AB - As part of a previous study, we identified Ontario cases of isocyanate-induced occupational asthma (OA) and the companies at which they worked. The Ontario Ministry of Labour maintained a computerized database including isocyanate air sampling determinations conducted by the Ministry. Within this database, we compared levels of isocyanate concentrations measured at 20 case companies [with compensated isocyanate asthma (OA) claims] with 203 noncase companies, based on air samples collected during the same 4-year period during which the OA claims arose. The proportion of case companies that were ever recorded as having a measured ambient isocyanate concentration of > or = 0.005 ppm was greater than that for noncase companies, for TDI users (43% vs 22%), and for MDI users (40% vs 27%). This reached conventional significance when combined across companies and isocyanate types (50% vs 25%; P < 0.05). This provides some evidence that facilities having OA claims have higher isocyanate exposures than do those without claims. PMID- 9327077 TI - No evidence for the influence of HLA class II in alleles in isocyanate-induced asthma. AB - Isocyanates are one of the main causes of occupational asthma. The aim of this investigation was to study the possible genetic background of isocyanate-induced asthma under consideration of the atopy status and different lung function parameters. We investigated the human leukocyte antigen (HLA) genes DRB1,3,4,5, DQB1, and DQA1 in 55 isocyanate-exposed patients with workplace-related dyspnea (32 asthmatics, 23 nonasthmatics) and 90 nonexposed controls. In contrast to other studies, we found no significant differences for any HLA class II allele tested in our study group. Furthermore, no significant differences concerning the aspartic amino acid residue 57 of DQB1 was observed. Therefore, we are unable to confirm an involvement of a specific HLA class II allele or DQB1-Asp57 in conferring susceptibility to isocyanate asthma in our study group. PMID- 9327078 TI - Occupational risk of Mycobacterium tuberculosis infection in hospital workers. AB - We conducted a 4-year (1/89-12/92) retrospective cohort study among employees at a large metropolitan hospital where a nosocomial outbreak of multidrug-resistant tuberculosis (TB) had occurred. We compared the risk of tuberculin skin test (TST) conversion among employees who worked on wards where patients with culture confirmed TB were cared for ("exposed") with the risk among employees who worked on wards with no such patients ("unexposed"). Exposed employees had a higher 4 year risk of TST conversion (14.5%) than unexposed employees (1.4%) (adjusted relative risk 13.4; 95 percent confidence interval 5.1-35.2). Exposed employees had significantly higher risks of conversion than unexposed employees during 1989 91, but not for 1992. Among the exposed, ward clerks had a risk of conversion (15.6%) only slightly lower than nurses (18.2%). We conclude that employees who worked in areas where patients with active M. tuberculosis infection were cared for, including workers who did not provide direct patient care, had a higher risk of TST conversion than employees who did not work in these areas. Reasons for the decline in risk over time include outbreak termination, fewer admissions of patients with TB, implementation of effective infection control measures, and possible resistance to infection in some members of the study population. PMID- 9327079 TI - Mortality in employees of a Scottish paper mill. AB - To assess possible health risks associated with the manufacture of paper, we carried out a retrospective analysis of mortality among 4,242 men and women employed at a Scottish paper mill between 1955 and 1992. During follow-up to 1994, 959 subjects had died giving an SMR of 0.85 (95% CI 0.80-0.90) in comparison with the national population. Mortality from all cancer (SMR 0.77, 95% CI 0.68-0.88) and particularly from lung cancer (SMR 0.64, 95% CI 0.50-0.81) was lower than expected. An excess of lymphatic and hematopoietic cancer (11 deaths, SMR 2.17) was observed in the making department. These findings do not support an occupational hazard of lung cancer as suggested by several earlier studies. The excess of lymphatic and hematopoietic cancer in the making department was unexpected, and may be a chance occurrence. PMID- 9327080 TI - Lead exposure in ironworkers. AB - In adults, lead toxicity is most commonly caused by occupation in a lead industry. Whereas lead toxicity has been described in workers who are involved in bridge rehabilitation, as of this date there has been no systematic evaluation published regarding the conditions responsible for lead toxicity in ironworkers. This is a report of a study designed to identify risk factors for elevated blood lead levels in ironworkers. One hundred fifty members of a 2,400-member local ironworkers union volunteered to have their blood drawn for lead and zinc protoporphyrin analysis and to complete a questionnaire regarding demographics, health, and occupation. The relationships between these variables and blood-lead level were analyzed using student's t-test, chi-square, and logistic regression. Current work on a lead job, rivet busting as the predominant job task, and cigarette smoking were all found to be significantly associated with elevated blood-lead level. Whereas cigarette smoking and current work with lead have been previously identified as risk factors for toxicity, interventions to prevent lead toxicity in ironworkers should also focus on work practices during rivet busting. PMID- 9327081 TI - Simple visual reaction time in organolead manufacturing workers: comparison of different methods of modeling lead exposure and reaction time. AB - In March 1990, 222 organolead manufacturing workers and 62 nonexposed referents were administered a neurobehavioral test battery that included simple visual reaction time (SVRT). SVRT was measured over 44 trials with interstimulus intervals ranging from 1 to 10 sec in a random but fixed order for all study subjects. Different measures of lead exposure and dose (e.g., recent and cumulative exposure based on personal sampling data, exposed/nonexposed status, recent blood lead and zinc protoporphyrin levels, and peak and cumulative urine lead levels) were examined as predictors of several different parameters of SVRT (e.g., mean, median, truncated mean, and standard deviation of SVRT over 44 trials). The association varied, depending on the measures used for SVRT and lead exposure and dose. In linear regression analyses, the strongest and most consistent associations of lead exposure and dose were observed with the standard deviation of SVRT. In assessing the different exposure measures, strong and consistent associations were observed with blood lead levels at the time of SVRT testing, but not with recent or cumulative exposure measures. That is, stronger associations were observed with measures of relatively recent internal dose (i.e., blood lead level) than with cumulative measures (i.e., cumulative exposure). Future studies using SVRT should consider parameters of SVRT that have not been commonly used to date, such as the standard deviation of the SVRT. PMID- 9327082 TI - Association of occupational and non-occupational risk factors with the prevalence of self-reported carpal tunnel syndrome in a national survey of the working population. AB - To compare the association of occupational versus personal, nonoccupational risk factors with the prevalence of carpal tunnel syndrome (CTS), data from the 1988 National Health Interview Survey, Occupational Health Supplement, were analyzed. When both occupational factors (bending/twisting of the hands/wrists [B/T] and use of hand-held vibrating tools) and personal nonoccupational factors (gender, race, age, body mass index [BMI], smoking, education, and family income) were included in a multivariate logistic regression model, adjusted odds ratios (AORs) of these factors for reporting medically called CTS (MC-CTS) were: exposure to B/T, 5.5; exposure to vibration, 1.9; white race, 16.7; female gender, 2.3; BMI > or = 25, 2.0; history of cigarette smoking, 1.6; age > or = 40, 1.2; education > 12 years, 1.2; and annual family income > or = $20,000, 1.5. Although both occupational and nonoccupational factors are associated with reporting of CTS, repetitive bending/twisting of the hands/wrists and use of vibrating tools remain important risk factors for work-related carpal tunnel syndrome. PMID- 9327083 TI - Male proportion in offspring of parents exposed to strong static and extremely low-frequency electromagnetic fields in Norway. AB - Reduced male proportion in offspring of male carbon setters prompted a study into whether offspring of workers exposed to strong static and extremely low-frequency electromagnetic fields (ELF) had a deviant sex ratio. The study was based on all births in Norway 1970-1993. The reference population was offspring of parents not exposed to ELF. The male proportion in offspring of men in aluminum works was 50.38%, RR 0.98 (0.94-1.03), in manganese works 47.32%, RR 0.92 (0.83-1.02), in factories producing iron 50.03%, RR 0.97 (0.93-1.02), in nickel works 48.27%, RR 0.94 (0.84-1.05), and in electric wire production 47.20%, RR 0.92 (0.80-1.05). In the offspring of women in aluminum works, the male proportion was 37.04%, RR 0.72 (0.59-0.90), in all smelter works grouped together, 45.13%, RR 0.88 (0.79-0.99). The male proportion in the reference population was 51.42%. The male proportion in offspring of men in industries with ELF, was slightly reduced, while offspring of women was significantly reduced. PMID- 9327084 TI - Professor Matthew Stewart: asbestosis research 1929-1934. AB - Matthew Stewart, Professor of Pathology at Leeds University, developed an interest in asbestosis during the late 1920s. In 1929, the Medical Research Council (MRC), encouraged by an advisory committee, funded research into asbestosis at Leeds University. Stewart supported by physicians designed a program of clinical, radiological and physiological studies to follow up Merewether's affected asbestos workers. Unfortunately, this met with opposition from industry, and the Home Office Factory Department was reluctant to assist, so it was abandoned. Industry did, however, cooperate with Stewart's studies on the effects of exposing guinea pigs in the factory environment, but this led to little in the way of publication. The failure of the Leeds School to realize its potential in investigating the effects of asbestos in humans, results in part from the discouragement it received and in part from the limited time and energies available to persons with a wide range of active interests. Some 45 years were to elapse before the MRC were enabled to carry out an analysis of the clinical, radiological and physiological data of a population of asbestos workers. PMID- 9327085 TI - Myelofibrosis in golf course groundskeepers. PMID- 9327086 TI - ApoE genotype and Alzheimer's disease in adults with Down syndrome: meta analysis. AB - The apoE gene polymorphism was examined in 100 adults with Down syndrome (with and without dementia) compared to 346 control subjects without mental retardation. Meta-analysis of available data (480 subjects) revealed that apolipoprotein E genotype distribution for people with Down syndrome was similar to that of the nonretarded population. Although no significant association between possession of the apoE epsilon 4 allele and onset of Alzheimer's disease was found, subjects with the allele had a tendency towards lower age of onset of dementia. Subjects with apoE epsilon 2 allele may not develop dementia and may have increased longevity. PMID- 9327087 TI - The quality of friendships between children with and without learning problems. AB - The nature and quality of preadolescent friendships between children with and without learning problems due to mental retardation or mild cognitive difficulties were investigated. Based on an assessment of the reciprocal relationship status of 373 children, including 54 with learning problems, 33 friend and 32 acquaintance dyads were identified. Of these dyads, half included a child with learning problems and half consisted of 2 children without learning problems. The dyads were observed performing a play task. Unlike friendships between children without disabilities, friendships between children with and without learning problems were marked by limited collaboration and shared decision-making, a low level of cooperative play and shared laughter, and an asymmetrical, hierarchical division of roles. The importance of advancing beyond the study of the social acceptance of children with learning problems to study the qualitative aspects of their friendships was discussed. PMID- 9327088 TI - Drug abuse in persons with mental retardation: a review. AB - The literature on drug abuse by people with mental retardation, a real, but largely ignored problem, was reviewed. Topics addressed were (a) the prevalence of drug use, (b) the drug-related problems characteristically encountered by this population, (c) the special vulnerabilities of people with mental retardation, (d) the treatment programs used with (and appropriate for) people with mental retardation, and (e) the status of drug abuse prevention and drug education for this population. Controlled research dealing with the genesis, treatment, and prevention of drug abuse among people with mental retardation is essentially nonexistent, but badly needed. PMID- 9327089 TI - The Stereotyped Behavior Scale for adolescents and adults with mental retardation. AB - The development of the Stereotyped Behavior Scale for adolescents and adults with mental retardation was described. Service provider staff in three states selected 600 clients known for stereotypic behaviors and then used 66 items to rate the frequency of occurrence of these behaviors. Items with test-retest reliability less than .45 and/or interrater agreement less than .30 were deleted. The remaining 30 items were subjected to a principal component analysis with varimax rotation. A single-factor solution emerged, which explained 24.9% of the variance. After eliminating 4 items with low factor loadings, we found that the final 26-item Stereotyped Behavior Scale had an internal consistency alpha of .88, test-retest reliability was pI = .90, and interrater reliability was pI = .76. PMID- 9327090 TI - Procedural and declarative memory processes: individuals with and without mental retardation. AB - Learning and retention of procedural versus declarative memory tasks were examined with 26 young adults with mild mental retardation and 27 school children matched for MA. Results revealed a similar pattern of task performance. Performance of the young adults with mild mental retardation was inferior to that of the control subjects on both types of tasks. However, learning rate and retention over time were comparable, thereby maintaining the control group's consistent advantage throughout all repeated trials. These results are consistent with previous findings for individual's with mental retardation tested on memory and problem-solving tasks. Theoretical implications of this pattern of results for individuals with mild mental retardation were discussed. PMID- 9327091 TI - Psychological distress of parents of infants with Down syndrome. AB - The distress level of parents who had infants with Down syndrome (study parents) was compared to that of control parents of infants without disability (infants were all less than 2 years of age). Data were collected in two independent surveys. We matched subjects case-by-case on socioeconomic status. Analysis of pooled data indicated significantly greater depression for the study parents. However, effect sizes were small, and the prevalence of clinical depression was 5.56% (n = 108) among matched study parents and 4.26% (n = 188) among unmatched study parents. Parenting an infant with Down syndrome may cause less distress than previously thought. PMID- 9327092 TI - Visual discrimination and motor reproduction of movement by individuals with mental retardation. AB - Visual discrimination and motor reproduction tasks involving computer-simulated arm movements were administered to 12 adults with mental retardation and a gender matched control group to examine whether inadequacies in visual perception account for the poorer motor performance of individuals with mental retardation. In the discrimination phase subjects judged whether simulated arm movements were either of greater or lesser extent or shorter or longer in duration, respectively, than those of a standard display. In the reproduction phase accuracy in reproducing the movement in the standard display was measured. Results indicate that error in discriminating extent and duration was significantly greater for the individuals with mental retardation, who were also less accurate and more variable in matching the extent and duration of the standard displays. These outcomes implicate both perceptual and motor reproduction inadequacies in skill acquisition for these individuals. PMID- 9327093 TI - Dissociation of POMC peptides after self-injury predicts responses to centrally acting opiate blockers. AB - Apparent insensitivity to pain, ritualistic patterns of behavior, and improvement in symptoms after administration of opiate receptor blockers implicated the endogenous opioid system in the initiation and maintenance of SIB. This study was designed to determine whether plasma levels of proopiomelanocortin (POMC)-derived peptides, beta-endorphin-like activity (beta E), ACTH, and adrenal cortisol immediately after an episode of SIB predicted subsequent response to an opiate blocker. Blood samples were collected from 10 patients with mental retardation within minutes of a self-injuring act and during an SIB-free control period. On another day, morning and afternoon samples were collected at least one week apart from the other samples. Effects on SIB of naltrexone hydrochloride (NTX) were examined in a double-blind, placebo-controlled crossover study. After an SIB episode, beta E, but not ACTH, was elevated compared with morning levels, p < .003. Patients with increased plasma levels of beta E after SIB had the most positive response to 2 mg/kg NTX, p < .03. Results suggest that changes in the hypothalamic-pituitary-adrenal axis after SIB may predict differences in individual patient response to opiate blockers. PMID- 9327094 TI - Western lowland gorillas (Gorilla gorilla gorilla) as seasonal frugivores: use of variable resources. AB - The gorillas studied at Bai Hokou, Central African Republic, between August 1990 and October 1992 consumed 239 kinds of foods from 138 species of plants and invertebrates, including the fruits of 77 species. Seeds were present in 99% of all fecal samples (n = 859). Although gorillas ate fleshy fruit whenever it was available, herbaceous plants and fibrous fruits were consumed year-round and were important during times of fleshy fruit scarcity. At Bai Hokou and across their range, resources are temporally discontinuous, and western gorilla diet exhibits marked seasonal and interannual variation. Although their large body size lends them dietary flexibility relative to chimpanzees, seasonal fruit-eating shapes the foraging and ranging patterns of western lowland gorillas. PMID- 9327095 TI - Ranging and grouping patterns of a western lowland gorilla group at Bai Hokou, Central African Republic. AB - The ranging and grouping patterns of a gorilla group were studied during 27 months from 1990-1992 at the Bai Hokou study site, Central African Republic. The study group ranged far daily (average = 2.3 km/day) and had a large home range (22.9 km2), relative to mountain gorillas, and ranging patterns differed between years. During 1990-1992, the bimale study group foraged less cohesively and had more flexible grouping patterns than mountain gorillas. The study group sometimes split into two distinct foraging subgroups, each led by a silverback, and these subgroups occasionally slept apart (mean = 950 m apart). Lowland gorillas rely on many of the same fruit resources as sympatric chimpanzees, and under certain demographic situations gorillas, like sympatric chimpanzees, may adapt their foraging group size to reduce intragroup feeding competition. However, the fiber content of the lowland gorilla diet likely relaxes constraints on foraging party size and facilitates group cohesion relative to chimpanzees. PMID- 9327096 TI - Social context affects phee call production by nonreproductive common marmosets (Callithrix jacchus). AB - Common marmosets produce two variants of their long call (phee call) in different situations. Intergroup calls are produced in territorial situations, and intragroup separation calls are produced by marmosets isolated from group members. Marmoset groups frequently include postpubertal, nonreproductive members; their roles in the spontaneous production of territorial vocalizations is unclear. This study analyzed the production of home cage phee calls by nonreproductive, postpubertal marmosets while they were housed in their natal groups and after pairing with an opposite-sex conspecific. Additionally, the production of the separation phee call variant was assessed in both social conditions. The results indicated that the marmosets rarely produced home cage, or territorial, phee calls while they were natally housed. In contrast, both males and females produced the territorial phee call at a much higher rate as early as 4 days after pairing. Agematched females remaining in their natal groups throughout the study produced home cage phee calls infrequently. Most marmosets produced separation phee calls at a high rate after separation from either their natal group or a partner, suggesting that the makeup of a social group has little effect on an animal's motivation to reunite with conspecifics. These results suggest that the social environment has an important influence on the production of territorial phee calls. PMID- 9327097 TI - Kin recognition by paternal half-siblings in captive Papio cynocephalus. AB - Our objective in this study was to evaluate whether a group of paternally related, subadult baboons (Papio cynocephalus) would preferentially interact with kin or nonkin when they had been raised apart from kin other than their mothers. Subjects and their mothers were removed from the breeding group and placed in alternate housing within 24 h after birth to ensure that the subjects would not have a social history with either their sire or their half-siblings. At 90 days of age, the 23 subjects were separated from their mothers and assigned to a peer peer social group. Behavioral performance was measured using focal animal sampling techniques and 12 molecular behavioral criteria. Analyses of the data indicate that in dyadic interactions kin did not interact more frequently than nonkin in performance of affiliative, sociosexual, and agonistic behaviors. The hypothesis that baboons recognize kin in the absence of maternal associations was not supported by the data; moreover, we suggest that social learning and social history are the most likely mechanisms for kin recognition. PMID- 9327098 TI - Meat-eating by adult female Sumatran orangutans (Pongo pygmaeus abelii). AB - Information about meat-eating behavior by wild orangutans (Pongo pygmaeus) is scant. The first article about such a case dates from 1981. Since 1989, seven incidents of adult female Sumatran orangutans eating slow lorises (Nycticebus coucang) have been witnessed. Three females from two study sites were involved. In three cases the females were seen catching the prey. There are too few cases to conclude whether this behavior is typically female. PMID- 9327100 TI - Albert Ellis on rational emotive behavior therapy. Interview by Lata K. McGinn. PMID- 9327099 TI - Group size and group composition of the mona monkey (Cercopithecus mona) on the Island of Grenada, West Indies. AB - Cercopithecus, the genus of guenons, is the largest of the African primate genera, and yet more than half of the species belonging to this group have never been the focus of a long-term field study. In this paper, I present data on group size and composition for a previously unstudied population of guenons on the Caribbean island of Grenada. The mona monkey, Cercopithecus mona, was introduced to Grenada from Africa approximately 200-300 years ago. Two types of social groups were found for Cercopithecus mona on Grenada: all-male groups consisting of two to four individuals and bisexual groups containing 5-32 individuals. All male groups of Grenada mona monkeys contained any combination of juveniles, subadults, and/or adults. All-male groups were a common occurrence on Grenada but have never been reported for African C. mona and have been reported only in two other forest Cercopithecus species. Bisexual groups appeared to consist of one adult male, one to six adult females, subadult females, and juveniles and infants of both sexes. Even though no more than one adult male was ever seen in each bisexual group of monas on Grenada, other males were heard giving copulation calls simultaneously with resident adult male loud calls, suggesting that other males occasionally infiltrate bisexual groups. PMID- 9327101 TI - Empathic understanding revisited: conceptualization, controversies, and limitations. AB - Regardless of the controversial philosophical aspects of understanding another human being, it is a useful concept in psychotherapy because understanding of the patient is one of the factors determining the course of therapy. Understanding resulting from empathy refers to the perception of the patient's innermost feelings and meanings, but inevitably represents only an approximation of what the patient truly experiences. It crucially depends on the vicissitudes of interactions between the patient and the therapist. The accuracy of empathic understanding needs to be verified continuously. It is important for therapists to learn to recognize what constitutes their own projection in their understanding of patients. Empathic understanding usually entails an attempt on the part of the therapist to feel to a certain extent the way the patient does. Various degrees of empathic emotional involvement may be suitable for patients with different types of psychopathology. Therapists must be careful as to how they convey their understanding to their patients. Not every patient wants to be understood, many are afraid of being understood, and some are distressed by the therapists' demonstration of understanding. Therefore, knowing what to do with the understanding one has achieved becomes an important therapeutic task. PMID- 9327103 TI - From object-relations theory to the theory of alterity: shame as an intermediary between the interpersonal world and the inner world of psychic structure. AB - Proceeding from a critical discussion of positions adopted in object-relations theory and of recent approaches to the understanding of shame (exemplified with reference to Wurmser's concept of shame), the paper demonstrates that intrapsychic structures should not be regarded as preconditions for shame but as themselves evolving in the first place from contact with experiential forms of the shame affect. The paper takes its theoretical bearings from object-relations theory and the theory of psychic structure, expanding the purview of these approaches by incorporating the reciprocity aspect and thus outlining a comprehensive "alterity theory." "Shame" is presented as an "interface affect," manifesting itself initially in the external interactional dimension and constituting the relational structure of "self-consciousness" via the internalization of the reciprocal relation between subject and object. From the angle of developmental psychology, three characteristic forms of such stages of internalization (identified by mythological figures) are described. (1) Narcissus, characterized by the absence of any reciprocal relation and accordingly termed "unreflected," (2) Tiresias, with a capacity for taking up the position of the vis-a-vis temporarily and looking critically at one's own self from that perspective, yet lacking the faculty of self-objectification without the help of the vis-a-vis. Self-objectification is therefore taking place in the interactional dimension. This stage is thus designated as "externally reflected." (3) Oedipus, who has reached the stage to be termed "self-reflected" or "self referential." The gaze is directed initially toward the outside in search of external sources of guilt but then falls back upon the subject itself. The gaze "turns inward" (to use an experientially suggestive image) and in the mythology this is represented by Oedipus' self-blinding. The subject is capable of "critical," dissociating functions, in the sense of self-objectification, thus attaining to a capacity for self-recognition, self-criticism and self-judgement. These three stages are seen to be progressive, not mutually exclusive. Self referentiality in the broadest sense is regarded as being hierarchically stratified. PMID- 9327102 TI - The impatient therapist: managed care and countertransference. AB - The conduct of therapy is particularly vulnerable to the influence of impatience. External pressures on the therapist to work quickly intrude on the session. When therapists are unconscious of their own impatience, it is more likely to reduce their ability to listen fully, to understand the extent of the patient's problems, and to participate in the rapidly shifting dance that makes therapy effective. Since the external factors that encourage this impatience are unlikely to abate, therapists need to become more aware of these influences and to consciously develop coping strategies. Impatience can arise from a number of places-the departure from any plan or timetable we have set for therapy, a desire to satisfy third parties, worry that the work is not progressing quickly enough, doubts about our ability to help, fear of "failure," confusion of process with "product," and attachment to favorite treatments. Aversion, attachment, and confusion have been well charted by Buddhist writers as hindrances to clear perception. The solution they propose is to face these obstacles directly, by noting their appearance, development, and passing away. While frequently an unpleasant process, such self-examination maximizes our chances of listening carefully and compassionately to each person who consults us. PMID- 9327104 TI - Exploitation of patients: themes in the psychopathology of their therapists. AB - This clinical paper develops an interpersonal theory of the therapist-patient relationship when boundaries are violated. It describes a clinically useful way to define boundary transgressions. A variety of mutually created dialectical paradigms interact with the therapist's psychopathology when, for various reasons, the therapist who is fully ethically responsible cannot see what is happening. The role of burgeoning aggression, dominance/ submission interplay, envy, and erotized countertransference is explored as forces leading to sexual and nonsexual exploitation of patients. Numerous clinical vignettes are used to illustrate the theoretical issues. Two fundamental axioms of treatment are also described, which, if followed, can help avoid transgressions. PMID- 9327105 TI - The clinical utilization of early childhood memories. AB - Early childhood memories (EMs) are discussed as an individual's unique psychological product, capable of revealing basic fantasies around which the individual's character structure is organized. Major theoretical approaches to understanding EMs are surveyed including early Freudian, Adlerian, and Ego Psychological. The more recent Cognitive-Perceptual perspective is also discussed. As EMs are viewed as a projective technique, methods of retrieval and interpretation are described. Case material is presented to demonstrate the utilization of EMs in the course of psychotherapy. In the early phase of therapy, EMs are shown to be of use in establishing therapeutic focus and elucidating dynamic patterns. Within the process of therapy, illustrations are given to demonstrate the use of EMs as an interpretive aid and as a means of monitoring therapeutic change. PMID- 9327106 TI - Threats to identity in survivors of multiple AIDS-related losses. AB - Survivors of multiple AIDS-related losses face threat to their identity because of the extreme disruption to their personal, assumptive, and interpersonal worlds. This article briefly explains the experience of multiple-loss survivors and includes a case history of a survivor. An individual's sense of self is transformed through identification with the disease. In the gay community, a particularly strong identification with AIDS arose. One outcome of the meshing of an AIDS and homosexual identity is the tendency for gays to assume an identity in relation to HIV ("I am HIV positive/negative.") Personality alteration is not uncommon and may include an inability to trust, labile emotionality, and diffuse anger. Erik Erikson's developmental stage model is used to clarify the confusion survivors face in maintaining and forming identity. Many survivors are catapulted into an integrity versus despair task, reporting many similarities with the situation of their grandparents. The survivor's interpersonal connection to the world, especially their connection to a community, is severely shaken. The article does not ignore the potential for positive identity growth arising from this tragedy. Conclusions from this experience may have applicability in other areas of multiple, ongoing losses. PMID- 9327107 TI - Change in the therapist: the role of patient-induced inspiration. AB - Clinicians experience positive changes in emotion, cognition, and behavior that they attribute to their work with particular patients. This process of change is termed inspiration. The goals of this study are: (1) to demonstrate that clinicians report being inspired by certain patients; (2) to compare inspiring and noninspiring patients regarding the strength of the therapeutic alliance; (3) to explore how the therapist changes as a result of working with the inspiring patient, and (4) to investigate what patient factors are inspiring. Of 300 randomly selected NASW members in the New York metropolitan area working in mental health settings, 84 completed a survey about inspiring and noninspiring patients that included the Working Alliance Inventory. Of these participants, 59 provided narrative responses about ways in which they have changed as a result of their work with inspiring patients. Inspiring patient factors were described by 37. The therapeutic alliance with the inspiring patient was found to be stronger than that with the noninspiring patient. Clinicians described various changes in personal and professional realms. In addition, specific patient factors such as perseverance and the ability to overcome serious obstacles were described as inspiring. The process of inspiration and implication of the findings in this study are discussed. PMID- 9327108 TI - Protein extract as a surrogate mother. AB - Brief dynamic psychotherapy, carried out by an resident in training, can bring significant improvement to a patient who does not necessarily meet ideal selection criteria. S. had experienced early childhood deprivation resulting in adult psychosomatic manifestations of feeling hypoglycemic under pressure of responsibility or during fear of abandonment. He soothed himself with the aid of a bottle of "protein extract," which he carried at all times on his belt. The 21 week therapy resulted in his increased ability to differentiate the sources of his anxiety and allowed him to find new and more mature ways of coping with stress. Although still confronting internal conflicts and anxiety, he relinquished the protein-extract bottle and began to take a more active role in work and relationships. With the help of brief dynamic psychotherapy, this patient was able to start moving forward in his personal growth. Finally, although requiring ongoing supervision and training, resident therapists should be encouraged to treat difficult patients with brief dynamic psychotherapy despite their limited experience with such a promising therapeutic technique. PMID- 9327109 TI - Psychodynamic psychotherapy with the older adult: challenges facing the patient and the therapist. AB - The subject of advanced age is often neglected in discussions of diversity and intersubjectivity in psychodynamic psychotherapy. In addition, psychologists historically have underestimated the ability of older individuals to utilize and benefit from psychodynamic treatment. This article provides support for the belief that many older individuals are capable of engaging in insight-oriented treatment and addresses some of the unique challenges faced by the older patient and the psychotherapist. The importance of determining the impact of age on intrapsychic conflicts is examined. Concepts from developmental psychodynamic theory are reviewed and applied to the clinical case of A., an 81-year-old widow. This case illustrates several themes that often emerge in work with older patients. The significance of changes in family structure and roles, object loss, and narcissistic injury in the life of this patient and older individuals in general is discussed. PMID- 9327110 TI - The evolving nature of psychotherapy outcome research. PMID- 9327111 TI - Interpersonal relationship and prisoner of war concerns of rated military male and female aircrew. AB - BACKGROUND: The issue of women flying military aircraft in a combat role has been very controversial. HYPOTHESIS: To succeed, female military aircrew are very similar to their male peers. METHODS: We conducted a comprehensive anonymous questionnaire survey of all U.S. Army and U.S. Air Force rated female aircrew, with an equal number of age and duty matched male aircrew. We are reporting on the interpersonal relationship and prisoner of war (P.O.W.) responses here. RESULTS: Male and female aircrew respond in a similar manner to posed questions, although differences do exist. Women reported: unequal treatment by opposite gender peers; problems relating to peers, superiors and subordinates; their gender influences assignments; the need to perform to higher standards and the need to work harder to be accepted as equals; ability to bond equally to their own and opposite gender peers; improved squadron cohesiveness in mixed gender squadrons; problems with peers' spouses; and, in a P.O.W. situation, fear of rape and sexual abuse. Men reported: women get inappropriate privileges and get special "breaks"; a gender difference in how flight duties are performed; worsened squadron cohesiveness in mixed gender squadrons; less likeliness to recommend their career path to their daughters; and a higher concern for welfare of families in a P.O.W. situation. CONCLUSIONS: Although responding in a similar manner to most questions, male and female military aircrew differ in the perception of their ability to function in mixed squadrons because of their gender. Some of these perceptions can be modified through training, others may need to be resolved through high level orders/policy; while in others, the military may have to accept women are different from men in some aspects. PMID- 9327112 TI - Slowing due to acute hypoxia originates early in the visual system. AB - BACKGROUND: Experiments in the visual modality show that acute hypoxia slows the earliest stage of information processing-preprocessing. It is unknown, however, whether a later stage, feature extraction, is also slowed. METHODS: To answer this question, an additive factors method (AFM) experiment was conducted which employed seven well trained subjects whose arterial oxyhaemoglobin saturation was controlled at 63% with low oxygen mixtures. The subjects responded to oddball names presented on a computer screen and both reaction time (RT) and the event related brain potential P300 were measured. The luminance and quality of the names was varied factorially to influence the preprocessing and feature extraction stages, respectively. RESULTS: RT and P300 latency showed the same pattern of results: stimulus luminance and signal quality were additive, indicating that AFM assumptions were met; hypoxia and stimulus luminance were interactive but hypoxia and signal quality were additive. CONCLUSION: In conjunction with other evidence, we interpret these results to indicate that the locus of slowing produced by hypoxia is largely at the preprocessing stage, at least in the visual modality. Slowing at the preprocessing stage can be explained by hypoxia shifting the function relating RT and stimulus luminance to the right by a constant amount. PMID- 9327113 TI - Comparison of marezine and dramamine in preventing symptoms of motion sickness. AB - BACKGROUND: The most common pharmacological agents for alleviating symptoms of motion sickness in the U.S. are over-the-counter antihistamines. Two example are dimenhydrinate (Dramamine) and cyclizine (Marezine). HYPOTHESIS: Dramamine and Marezine suppress overall motion sickness symptoms with equal effectiveness, but Dramamine affects the central nervous system (CNS), while Marezine affects the stomach directly. METHODS: This study employed a double-blind, within-subject design to compare the effectiveness of Marezine (50 mg) and Dramamine (50 mg), in preventing subjective symptoms and gastric dysrhythmias associated with motion sickness. The sedative effects of the two drugs were also compared. Electrogastrograms (EGGs) were recorded from 23 subjects during 2 counterbalanced sessions for 3 trial periods: an 8-min pre-drug baseline, an 8-min pre-rotation baseline, which began 30 min after drug ingestion, and a 16-min period of exposure to a rotating optokinetic drum. Subjects reported any subjective symptoms of motion sickness (SSMS) and drowsiness before and during induction of motion sickness. RESULTS: There were no statistically significant differences between the two drug conditions for the overall mean SSMS scores. However, when the scores were divided into symptom groups, Marezine was associated with significantly lower scores than Dramamine for gastrointestinal (GI) symptoms. Also, Marezine was associated with significantly less drowsiness than Dramamine 30 min after ingestion. Power in both the normal (3 cpm) and tachyarrhythmia (4-9 cpm) ranges of the EGG increased significantly more during rotation compared to baseline in the Dramamine condition than in the Marezine condition. CONCLUSIONS: Marezine and Dramamine are similarly effective in preventing the overall subjective symptoms of motion sickness. While Dramamine's effectiveness may be related to its sedative properties, Marezine may work more directly on the stomach and thus be more effective in preventing gastric dysrhythmias and reports of GI symptoms. PMID- 9327114 TI - Are the laboratory and field conditions observations of acute mountain sickness related? AB - In order to study relationships between acute mountain sickness (AMS) observations done both during a short-term hypoxic exposure in a hypobaric chamber, and in field conditions during a high altitude expedition, nine subjects were submitted to a 9-h hypoxic exposure in a hypobaric chamber. Then, they experienced a high altitude expedition in the Himalayas. The Lake Louise AMS scoring system was used to assess AMS in both conditions, especially the self report questionnaire. During the expedition, the mean self report score of each subject, defined as the ratio between the sum of daily self report scores and the duration of the expedition, appears to be correlated not only to the maximal self report score observed in altitude (r = +0.77, p < 0.05) but also to the self report and self report+clinical assessment scores observed at the end of the hypobaric chamber sojourn (r = +0.81, p < 0.01 and r = +0.75, p < 0.05, respectively). In conclusion, the Lake Louise AMS scoring system, especially the self report questionnaire, is relevant to assess AMS with simplicity and rapidity both in laboratory and in field conditions. Our study also suggests that AMS induced by a short term sojourn in a hypobaric chamber is related to AMS observed in field conditions. PMID- 9327115 TI - Anti-retroviral therapy and cognitive function. AB - BACKGROUND: The effect of anti-retroviral medications on the cognitive functions important in flying has not been determined. HYPOTHESIS: Anti-retroviral medications have no effect on the cognitive performance of individuals at the CDC 4C2 (symptomatic HIV disease with no illness indicative of full-blown AIDS) stage of infection. METHODS: A two-group study using a cross-sectional design was used. The participants in each group represented a sample of convenience obtained from a larger, naturalistic study. Each group consisted of 10 HIV+ males at the CDC 4C2 stage of infection. The two groups were found to be comparable on age, education, pre-morbid intelligence, and ethnicity. All members of the anti retroviral medication group had been receiving medications for at least 3 mo. Those in the control group (no anti-retroviral medication) had received no anti retroviral medications for at least 6 mo. Cognitive functioning was assessed using a computerized information processing battery that included test similar to those under consideration for inclusion in military pilot selection batteries and a neuropsychological battery. As part of the larger study, the participants were carefully and repetitively screened on factors known to affect performance on neuropsychological instruments. RESULTS: The groups showed little difference in cognitive functioning. CONCLUSION: Although more research is needed, anti retroviral medication does not impair, and may improve, the cognitive processes of individuals with symptomatic HIV infection who do not have AIDS. PMID- 9327116 TI - An examination of two emergency breathing aids for use during helicopter underwater escape. AB - BACKGROUND: There is a paucity of published work in which the performance of Emergency Underwater Breathing Aids (EUBA) has been examined in the wide range of scenarios in which helicopter underwater escape may be necessary. In the present investigation two EUBA were examined: the Air Pocket (AP) rebreather and the Short Term Air Supply System (STASS) mini SCUBA set. METHOD: Young, healthy male subjects undertook simple simulated helicopter underwater escapes in water at 15 degrees C and/or 5 degrees C. During the immersions the subjects attempted to remain submerged for 60 s while traversing back and forth along a ladder secured at a depth of 1.25 m. At each temperature the subjects used AP and STASS twice. The subjects were dressed in the Royal Navy winter sea helicopter aircrew equipment assembly and an aircrew helmet. RESULTS: Both AP and STASS significantly extended the underwater survival time of individuals when compared to their maximum breath-hold time (BHT). It is clear from the measurements made of gas concentrations in AP; the volume of air used from STASS; and subjective responses, that the 60-s submersions were achieved more easily with STASS than AP. CONCLUSION: It is concluded that in conditions similar to those of the present experiment STASS will give a longer underwater duration than AP, but this benefit must be offset against the possible risk of pulmonary barotrauma associated with the use of STASS, as well as increased training and maintenance costs. Irrespective of the EUBA which is provided, in-water training, preferably including exposure to cold water, will significantly improve the ability of an individual to use it. PMID- 9327117 TI - Effects of weight and center of gravity location of head-supported devices on neck loading. AB - A comprehensive study of the effect of Head-Supported Devices (HSDs) on neck loading during helicopter accidents is presented. The new Articulated Total Body (ATB) model which treats the neck as a deformable segment was used for crash simulation. Different categories of human and manikin subjects were considered under several crash scenarios. Simulations were theoretically designed to include a wide range of HSDs by changing their weights and center of gravity (CG) locations relative to the head, and studying the effects of these changes. Since HSDs were only theoretically included in the model, detachment of specific detachable devices used in most military applications was not modeled in the study. Hence, two typical detachable devices were modeled and selected simulations were repeated and compared not only to provide a measure of accuracy for the original results but also to see the effect of separation of these devices from the helmet. PMID- 9327118 TI - The legal implications of preflight medical screening of civil airline passengers. AB - BACKGROUND: It has been suggested that meticulous preflight medical screening of airline passengers would prevent most in-flight medical emergencies and it has been estimated that medical assistance is sought on around 1 in 50 international flights on wide bodied domestic aircraft. It was considered that the legal implications of such screening needed to be determined. METHODS: A literature review of current legislation, court cases, and legal and medical journals was conducted. RESULTS: It was found that the legal problems with preflight medical screening fell into three areas: discrimination, right to free movement, and guidelines to medical contraindications to flying. It was considered that precluding someone from flying on medical grounds could in certain circumstances be construed as discriminatory or a breach of the basic human right of freedom of movement and, thus, unlawful. Current guidelines on medical contraindications to flying vary and there are presently no internationally agreed or legally enforceable protocols on the subject. CONCLUSIONS: Pre-flight medical screening of civil airline passengers may offer a means of reducing in-flight morbidity, but the complexity of the legal issues involved are such that it is unlikely to be introduced in the near future. PMID- 9327119 TI - Zolpidem as a fatigue countermeasure. AB - BACKGROUND: In light of greater emphasis on global reach, fatigue has a real potential to negatively impact the U.S. Air Force (USAF) mission. Sustained operations and extended air missions will become ever more common and critical in a smaller USAF and human performance may be the most important limiting factor in the effectiveness of advanced weapons systems. Pharmaceuticals may be considered for fatigue countermeasures. METHODS: A literature review was conducted to examine the potential of zolpidem as a pharmaceutical countermeasure against fatigue in USAF operations. RESULTS: Zolpidem is a hypnotic which appears to cause less global impairment than benzodiazepines during peak effect, and is free of persistent performance decrement or hangover effect. Few adverse effects have been reported. Several studies suggest a benefit of effective sleep produced by hypnotics on next day performance compared with sleep of questionable quality using no medication. CONCLUSION: The USAF should conduct further investigation into the potential safety and efficacy of zolpidem during sustained operations and extended air operations. PMID- 9327120 TI - Fatigue in the aviation environment: an overview of the causes and effects as well as recommended countermeasures. AB - Fatigue is an insidious threat to aviation safety because of the impairments in alertness and performance it creates. The fatigue associated with sleep loss, shift work, and long duty cycles can cause aviators to become sloppy, inattentive, careless, and inefficient. The only cure for fatigue is adequate sleep; however, gaining sufficient amounts of sleep is often difficult because of work requirements, family demands, or poor sleep habits. Although it may not be possible to avoid some of these problems, pilots can improve their sleep habits and thus gain more restful and restorative sleep by using self-administered relaxation therapy, establishing consistent and soothing bedtime routines, and avoiding certain activities and substances immediately prior to sleep. When opportunities for adequate sleep are not available because of work-related factors, prophylactic naps can sustain performance until sleep is possible. PMID- 9327121 TI - Human subject screening: a dynamic process. AB - INTRODUCTION: The history of disqualified (DQ) subjects from 1973-1993 at Armstrong Laboratory, Wright Patterson AFB, is presented for both sustained and impact acceleration panels. METHODS: Candidate and subject medical records were reviewed for screening results, recommendation for panel duty, and any follow-up medical findings. The generation and interpretation of the medical screening criteria and DQ rates are discussed. MEDICAL SCREENING CRITERIA: The mechanisms for change, those factors influencing change, and the interpretation of the screening criteria for Armstrong Laboratory's acceleration panels determine the panel's composition, which is reflected in the DQ rates. RESULTS: The centrifuge had a 5% (7/132) disqualification (DQ) rate from 1973-93 with 29% (2/7) due to musculoskeletal and 71% (5/7) for medical reasons. All were DQ during 1973-88. The impact panel had a DQ rate of 18% (36/195) with 71% (24/34) DQ due to musculoskeletal and 29% (10/34) for medical reasons. Only 28% (10/36) were DQ during 1973-88, while during 1989-93, 72% (26/36) were DQ. CONCLUSIONS: The differences in DQ rates between the centrifuge and impact facility were due to the variability or conservatism of individual physicians, interpretation of the medical screening criteria, and the type of research being done. These factors effect the composition of the human subject panels. This determines to which target population the research data can be applied. If the subjects do not represent pilots due to inappropriate screening, then there is no benefit from the research and, therefore, there can be no risk incurred by the subjects. PMID- 9327122 TI - The very large airplane: safety, health, and comfort considerations. Air Transport Medicine Committee, Aerospace Medical Association. AB - In recent years, aircraft manufacturers have been considering a very large airplane with a capacity of 600-1000 passengers. The human factors aspects of such an unprecedented enterprise demand that the aerospace medicine community take an active role early on in the design phase. Consequently, the Aerospace Medical Association formed an international task force to prepare a paper containing pertinent human factors recommendations for the manufacturers. This paper, including the recommendations herein, has been forwarded to Boeing and Airbus as well as to 50 major airlines of the world. PMID- 9327123 TI - Identification of preferred distamycin-DNA binding sites by the combinatorial method REPSA. AB - The combinatorial method restriction endonuclease protection, selection, and amplification (REPSA) was used to determine the preferred duplex DNA binding sites of the peptide N-methylpyrrolecarboxamide antibiotic distamycin A. After 12 rounds of REPSA, several sequences were identified that bound distamycin with an apparent affinity of 2-20 nM. Among these, the highest-affinity sites averaged 10 bp in length, suggesting that these sites may be occupied by multiple, cooperatively interacting distamycin molecules. Presently, REPSA is the only combinatorial approach that allows the identification of preferred DNA targets for small molecule ligands at physiologically relevant concentrations in solution. As such, it should prove useful in the design and screening of sequence specific DNA-binding molecules. PMID- 9327124 TI - 99mTc labeling of highly potent small peptides. PMID- 9327125 TI - Protein conjugates with water-soluble poly(alkylene oxide)s entrapped in hydrated reversed micelles. AB - Conjugates of alpha-chymotrypsin (ChT) with poly(ethylene glycol) (PEG) and block copolymers of ethylene and propylene oxides (proxanols) have been synthesized. The molecular mass of the polymers used was 2 kDa. The conjugates contained five to seven polymer chains per enzyme molecule. Hydrolysis of N-trans cinnamoylimidazole catalyzed by the conjugates of ChT with poly(alkylene oxide)s was studied in 0.05 M Tris-HCl buffer at pH 8.0 and in the system of the hydrated reversed micelles of aerosol OT (AOT) in octane at 25 degrees C. The deacylation rate constant k3 for the conjugates in buffer solution was 1.5-1.8-fold higher than that for native ChT. The value of the [H2O]:[AOT] ratio corresponding to the maximum on k3 versus [H2O]:[AOT] curves for the conjugates (ca. 16) allows the dimensions of their molecules to be evaluated. The radius of the conjugate molecules was found to be about 2.8 nm. The value of k3 for the conjugate of ChT with PEG, as in the case of native ChT, remains constant when the concentration of AOT is varied. However, the deacylation rate constant for the conjugates of ChT with proxanols decreases with the increase in AOT concentration, which indicates that these conjugates are able to interact with the micellar matrix and therefore may be considered membranotropic compounds. PMID- 9327126 TI - Induced thermostability of poly(ethylene oxide)-modified hemoglobin in glycols. AB - The thermostability and redox activity of poly(ethylene oxide) (PEO)-modified human hemoglobin in PEO200 (PEO containing KCl, average MW of 200, < 0.3% H2O) were investigated by UV-vis spectroscopy, by circular dichroism spectroscopy, and by cyclic voltammetry. Using PEO oligomers as a solvent, PEO-modified hemoglobin was reduced and oxidized at an indium tin oxide glass electrode in the temperature range of -10 to 120 degrees C. The thermostability of PEO-modified hemoglobin was affected by the molecular weight of the solvent PEO. In lower molecular weight glycols (MW of < 150), PEO-modified hemoglobin was denatured within a few minutes at 80 degrees C. On the other hand, the absorbance at the Soret band for PEO-modified hemoglobin was unchanged for 2 h at 80 degrees C in PEO200. A decrease in the water content of solvent PEO200 also improved the thermostability of PEO-modified hemoglobin. Improvement in the thermostability was attributed to physicochemical characteristics such as the relatively low molecular motion of PEO oligomers used as a solvent. PMID- 9327127 TI - Interactions of pluronic block copolymers with brain microvessel endothelial cells: evidence of two potential pathways for drug absorption. AB - Pluronic block copolymers have been previously reported to increase the delivery of agents to the brain [Kabanov et al. (1992) J. Controlled Release 22, 141-158]. In the present study, primary cultured bovine brain microvessel endothelial cells (BBMEC) were used as an in vitro model of the blood-brain barrier to examine the membrane interactions of Pluronic P85 (P85) and potential mechanisms for drug absorption. At concentrations below the critical micelle concentration (cmc), P85 enhanced the accumulation of the fluorescent probe rhodamine 123 (R123) in BBMEC through inhibition of P-glycoprotein (P-gp)-mediated drug efflux. The effects of P85 on the cellular accumulation of R123 were also observed in KBv cells (P-gp positive) but not in human umbilical vein endothelial cells (P-gp negative). In contrast to the effects with P85 below the cmc, the enhanced absorption of R123 observed with Pluronic micelles was transient and not dependent on P-gp. A transient increase in R123 accumulation was observed in both P-gp positive cells (brain microvessel endothelial cells and KBv) and P-gp negative cells (human umbilical vein endothelial cells). Therefore, it appears that P85 affects the absorption of drugs in brain microvessel endothelial cells through (1) inhibition of the P-gp-mediated drug efflux at low concentrations of the copolymer and (2) increased vesicular transport at higher concentrations of the copolymer. Furthermore, both interactions of P85 with the brain endothelial cells appear to be energy-dependent as demonstrated by the inhibitory effects of the metabolic inhibitor 2-deoxyglucose. PMID- 9327128 TI - Formation of microscale gradients of protein using heterobifunctional photolinkers. AB - Gradients of biological molecules on a microscale have been postulated to elicit cellular responses, such as migration. However, it has been difficult to prepare such gradients for experimental testing. A means for producing such gradients has been developed using a heterobifunctional photolinking agent with laser light activation. The photolinking agent synthesized includes an N-hydroxysuccinimide group and a photoreactive benzophenone (BP) separated by a tetraethylene glycol (TEG) spacer. The presence of the tetraethylene glycol spacer renders the photolinker hydrophilic, a desirable trait for conjugation in aqueous solutions. The linker was then conjugated to R-phycoerythrin (R-PE), a fluorescent protein. The resulting photolinker-R-phycoerythrin conjugate (BP-TEG-PE) was then immobilized onto a polystyrene surface by laser irradiation on a motorized stage. By varying exposure time of the sample to the beam, the amount of BP-TEG-PE immobilized on the surface was changed over an order of magnitude over a distance of 250 microns. This method can be applied to prepare gradients of proteins that elicit biological responses, such as extracellular matrix proteins or growth factors, and to study the biological effects of such gradients. PMID- 9327129 TI - Glucose-induced release of glycosylpoly(ethylene glycol) insulin bound to a soluble conjugate of concanavalin A. AB - Treatment of diabetes mellitus by insulin injections provides long-term control of the disease but lacks any feedback response to glucose concentration changes, which finally leads to a number of life-threatening conditions. The purpose of this study was to improve and optimize an implantable, concanavalin A (Con A) based, glucose-responsive insulin delivery system studied earlier [Jeong, S. Y., Kim, S. W., Holmberg, D. L., and McRea, J. C. (1985) J. Controlled Release 2, 143 152], which can be used for long-term diabetes treatment. To optimize the "insulin component" of the delivery system, we prepared PheB1 insulin amino group monosubstituted monoglucosylpoly(ethylene glycol) (G-PEG) insulin conjugates (PEG M(r) 600 or 2000), which showed preserved bioactivity, significantly improved solubility and solution stability at neutral pH, and substantially suppressed hexamerization/dimerization. To improve the delivery system further, we synthesized and characterized a conjugate of Con A and monomethoxypoly(ethylene glycol) (mPEG, M(r) 5000) grafted hydrophilic poly(vinylpyrrolidone-co-acrylic acid) (PVPAA) with M(r) of 250,000. The optimal conjugate contained around eight PEG chains and two to three Con A tetramers attached through the amide bonds to the PVPAA chain. The Con A sugar binding characteristics were preserved, and, more importantly, Con A solubility at pH 7.4 substantially increased. This also holds true for a complex formed by the Con A conjugate and G-PEG insulin, which is soluble and does not precipitate under the physiologically relevant conditions under which the complex formed by the Con A conjugate and glycosyl insulin immediately precipitates. Finally, no leakage of the Con A conjugate from a membrane device was detected. Preliminary in vitro release experiments with Con A conjugate and G-PEG insulin complex enclosed in the membrane device showed a pulsative, reversible release pattern for G-PEG insulin in response to glucose challenges of 50-500 mg/dL, demonstrating the feasibility of the release system for use in planned, chronic in vivo studies with diabetic (pancreatectomized) dogs. PMID- 9327130 TI - Design and synthesis of [111In]DTPA-folate for use as a tumor-targeted radiopharmaceutical. AB - Folate-conjugated metal chelates have been proposed as potential imaging agents for cancers that overexpress folate receptors. In a previous study, folic acid was linked through its gamma-carboxyl group to deferoxamine (DF), and the 67Ga labeled complex ([67Ga]DF-folate) was examined for in vivo tumor targeting efficiency in athymic mice with a human tumor cell implant. Although superb tumor to-background contrast was obtained, slow hepatobiliary clearance would compromise imaging of abdominal tumors such as ovarian cancer. In the present study, folic acid was conjugated to an alternative chelator, diethylenetriaminepentaacetic acid (DTPA), via an ethylenediamine spacer. The desired DTPA-folate (gamma) regioisomer was synthesized by two different approaches, purified by reversed phase column chromatography, and characterized mainly by analytical HPLC, mass spectroscopy, and NMR. In cultured tumor cells, uptake of [111In]DTPA-folate (gamma) was found to be specific for folate receptor bearing cells, and the kinetics of uptake were similar to those of free folate and other folate-conjugated molecules. In the normal rat, intravenously administered [111In]DTPA-folate (gamma) was found to be rapidly excreted into the urine, giving intestinal levels of radiotracer 10-fold lower than those observed with [67Ga]DF-folate (gamma) at 4 h. In a preliminary mouse imaging study, a folate receptor-positive KB cell tumor was readily visualized by gamma scintigraphy 1 h following intravenous administration of [111In]DTPA-folate (gamma). PMID- 9327131 TI - Photoaffinity analogue for the anti-inflammatory drug alpha-trinositol: synthesis and identification of putative molecular targets. AB - alpha-Trinositol (alpha T), or Ins(1,2,6)P3, is a semisynthetic inositol trisphosphate produced commercially by the partial degradation of phytic acid with phytase. The molecular targets mediating the mechanism of action of this novel anti-inflammatory, analgesic, and antivasoconstrictive drug are unknown. A new photoaffinity analogue, 4-[3H]BZDC-alpha T, has been prepared in which the [3H]-p-benzoyldihydrocinnamoyl ([3H]BZDC) photophore is tethered through an O-(5 aminopentanoyl) linkage to the 4-OH of alpha T. Photoaffinity labeling experiments with two human tissues, umbilical cord vascular smooth muscle cells and platelet membranes, revealed proteins that were selectively labeled by 4 [3H]BZDC-alpha T. Thus, co-incubation with alpha T but not with Ins(1,3,4,5)P4 during photolysis competitively displaced labeling of a 55 kDa platelet protein. In vascular epithelial cells, alpha T and Ins(1,3,4,5)P4 both displaced labeling of a 55 and 43 kDa proteins. The identification of putative protein targets for alpha T in smooth vascular tissue may have important implications in elucidation of the mechanism of action of this unusual drug. PMID- 9327132 TI - Synthesis and in vitro degradation of new polyvalent hydrazide cross-linked hydrogels of hyaluronic acid. AB - New polyvalent hydrazide cross-linkers were synthesized, characterized, and used to prepare hydrazide cross-linked hydrogels derived from hyaluronic acid (HA). First, the chemical synthesis and characterization of the di-, tri-, tetra-, penta-, and hexahydrazides are presented. Second, HA concentration, buffer type and concentration, and ratio of HA to carbodiimide to cross-linker were varied to obtain HA-hydrogels with different chemical and physical properties. Third, two new assays are described to monitor the stability of HA-hydrogels toward hyaluronidase (HAse) and other media. These assays were used to evaluate the stability of cross-linked HA-hydrogels to HAse solutions and different pH values. Hydrophobic cross-linkers gave the most stable gels, and the susceptibility of the gels to HAse was independent of cross-linker concentration. HAse does not significantly penetrate the HA-hydrogels and acts primarily at the gel-solution interface. The HA-hydrogels are stable in acid environments and dissolve gradually above pH 7.0. PMID- 9327133 TI - Genetic construction and characterization of an anti-monkey CD3 single-chain immunotoxin with a truncated diphtheria toxin. AB - We have previously developed a chemically conjugated anti-rhesus monkey CD3 immunotoxin FN18-CRM9 that can deplete in vivo T cells and induce long term tolerance of mismatched renal allograft in rhesus monkeys. This immunotoxin is a monkey analogue of anti-human CD3 immunotoxin UCHT1-CRM9. In this study, we cloned the light and heavy chain variable regions of anti-monkey CD3 monoclonal antibody FN18 and constructed a single-chain Fv (sFv) by linking variable light and variable heavy regions with a (Gly4Ser)3 linker. The single-chain immunotoxin DT390-FN18sFv was constructed by ligating the sFv to the carboxyl terminus of DT390, a truncated form of diphtheria toxin. The DT390-FN18sFv fusion protein was expressed in Escherichia coli and purified with Ni-RTA affinity and anion exchange columns. Similar to the chemically conjugated immunotoxin FN18-CRM9, DT390-FN18sFv can also specifically inhibit protein synthesis in primary monkey T cells in a dose-dependent manner. DT390-FN18sFv at 10(-7) mol/L or FN18-CRM9 at 10(-8) mol/L is sufficient to reduce protein synthesis of monkey primary T cells to less than 5% of the control. The 50% inhibition dosage (IC50) of FN18-CRM9 is 1 x 10(-10) mol/L, while the IC50 of DT390-FN18sFv is 1 x 10(-8) mol/ L, reflecting the lowered affinity of monovalent Fab' FN18 to its parental divalent antibody. The availability of functional FN18sFv will provide the basis for the construction of divalent anti-CD3 immunotoxins for preclinical studies on the induction of tolerance in organ transplantation and experimental autoimmune diseases. PMID- 9327134 TI - Water-soluble polyion complex associates of DNA and poly(ethylene glycol)-poly(L lysine) block copolymer. AB - Complex formation of poly(ethylene glycol)-poly(L-lysine) (PEG-PLL) AB type block copolymer with salmon testes DNA or Col E1 plasmid DNA in aqueous milieu was studied. The PLL segment of PEG-PLL interacts with nucleic acid through an electrostatic force to form a water-soluble complex associate with a diameter of ca. 50 nm. PEG segments surrounding the core of the polyion complex prevented the complex from precipitation even under stoichiometric conditions, at which the unit ratio of L-lysine in PEG-PLL and phosphate in the DNA is equal. The profile of the thermal melting curve revealed a higher stabilization of DNA structure in PEG-PLL/DNA complexes compared to that in the complex made from DNA and PLL homopolymer with the same molecular weight as the PLL segment in PEG-PLL. This stabilizing effect on the DNA structure may be due to the compartmentalization of DNA into the microenvironment of PEG with low permittivity. The reversible nature of the PEG-PLL/DNA complex was further verified through the addition of polyanion [poly-(L-aspartic acid)]: Poly(L-aspartic acid) replaced DNA in the complex with PEG-PLL, resulting in the release of free DNA in the medium. Furthermore, the PEG PLL/DNA complex showed high resistance against DNase I attack, suggesting DNA protection through the segregation into the core of the associate having PEG palisade. PMID- 9327135 TI - Expression and characterization of bryodin 1 and a bryodin 1-based single-chain immunotoxin from tobacco cell culture. AB - Bryodin 1 (BD1) is a potent ribosome-inactivating protein (RIP) isolated from the plant Bryonia dioica. It is relatively nontoxic in rodents (LD50 > 40 mg/kg) and represents a potential improvement over other RIPs and bacterial toxins that have been used in immunotoxins. Recombinant BD1, expressed in Escherichia coli, localizes to insoluble inclusion bodies necessitating denaturation and refolding steps to generate active protein. In this report, BD1 was expressed as a soluble recombinant protein in tobacco cell culture (ntBD1) and purified to near homogeneity with yields of up to 30 mg/(L of culture). The protein synthesis inhibition activity of ntBD1 was identical to that of both native BD1 isolated from the roots of B. dioica and recombinant BD1 expressed in E. coli. Toxicology analysis showed that ntBD1 was well tolerated in rats at doses that cannot be achieved with most other toxin components of immunotoxins. Additionally, a single chain immunotoxin composed of BD1 fused to the single-chain Fv region of the anti CD40 antibody G28-5 (ntBD1-G28-5 sFv) was expressed in tobacco tissue culture as a soluble protein and was specifically cytotoxic toward CD40 expressing non Hodgkin's lymphoma cells in vitro. These data indicate that tobacco tissue culture is a viable system for soluble expression of BD1 and BD1-containing immunotoxins. PMID- 9327136 TI - Synthesis and biological properties of mannosylated starburst poly(amidoamine) dendrimers. AB - Starburst PAMAM dendrimers ending with mannopyranoside residues were readily synthesized in large scale and good yields from commercially available dendrimers bearing high-density amine functionality on their surface and p isothiocyanatophenyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranoside. The first four generations of this novel class of monodispersed neoglycoconjugates having up to 32 mannoside units were evaluated as ligands for the phytohemagglutinins from concanavalin A (Con A) and Pisum sativum (pea lectin) using enzyme-linked lectin assay (ELLA) and turbidimetric analyses. The binding properties of these glycodendrimers, together with reference monosaccharides, were determined using yeast mannan as a coating antigen and peroxidase-labeled lectins. These mannosylated dendrimers were demonstrated to be potent inhibitors with IC50 values 400 times better than those of monomeric methyl alpha-D-mannopyranoside taken as a standard. Their lipophilic character was shown to be sufficient for their direct use as coating antigens in microtiter plate assays. Moreover, their ability to bind and form insoluble carbohydrate-lectin complexes was also demonstrated by radial double immunodiffusion and turbidimetric analyses. Furthermore, the ability of these ligands to selectively precipitate a mannose binding protein (Con A) from a crude lectin mixture was also demonstrated using polyacrylamide gel electrophoresis (SDS-PAGE). These multivalent neoglycoconjugates were shown to constitute novel biochromatography materials of high affinity for the easy isolation of carbohydrate-binding proteins. PMID- 9327137 TI - Method for radioiodination of proteins using N-succinimidyl 3-hydroxy-4 iodobenzoate. AB - A conjugation method has been developed for the radioiodination of proteins which should be adaptable to kit formulation. m-Hydroxybenzoic acid was converted to 3 hydroxy-4-[131I]iodobenzoic acid in 65% radiochemical yield using Chloramine-T as the oxidant. This intermediate was then converted to N-succinimidyl 3-hydroxy-4 [131I]iodobenzoate ([131I]mSHIB) in 75% yield by reaction with N hydroxysuccinimide and dicyclohexylcarbodiimide in a reaction time of only 10 min. Monoclonal antibody (mAb) 81C6 was labeled in 40-60% yield by reaction with [131I]mSHIB. Performing purifications of radioiodinated compounds using cartridges instead of HPLC did not alter conjugation efficiency, mAb immunoreactivity, or tissue distribution. Thyroid uptake of labeled mAb was low but up to 2.4 times higher than that seen when the mAb was labeled with N succinimidyl 3-[125I]-iodobenzoate. These results suggest that [131I]mSHIB may be a useful reagent for the radioiodination of proteins, particularly in contexts when less complicated purification methods would be advantageous. PMID- 9327138 TI - Photoimmobilization of sulfated hyaluronic acid for antithrombogenicity. AB - Anticoagulant polymer sulfated hyaluronic acid was patterned immobilized on a poly(ethylene terephthalate) (PET) film in a specific pattern by photolithography. Hyaluronic acid was sulfated by a sulfur trioxide-pyridine complex. The polymer was coupled with azidoaniline. The derivatized polymer was cast on a PET film from aqueous solution. After drying, the film was photoirradiated in the presence or absence of a photomask. The micropatterning was confirmed by staining with a dye, brilliant green. Since the anticoagulant polymer has negative charges, the cationic dye was adsorbed on the regions where the anticoagulant polymer was immobilized. Platelet adhesion was reduced on the sulfated hyaluronic acid-immobilized areas. The immobilized sulfated hyaluronic acid significantly reduced thrombus formation. PMID- 9327139 TI - Nanoparticle DNA carrier with poly(L-lysine) grafted polysaccharide copolymer and poly(D,L-lactic acid). AB - Biodegradable nanoparticles, which contain the sites for both polynucleotide adsorption and targeting ligand on their surfaces, were prepared as a novel carrier for genetic materials. The nanoparticles were obtained from poly(D,L lactic acid) and poly(L-lysine)-graft-polysaccharide copolymers by using either a solvent evaporation method or a diafiltration method. The size of the particles prepared by the diafiltration method was controlled by varying the initial concentration of the graft copolymer. Nanoparticles as small as 60 nm in diameter were successfully obtained from the graft copolymers with high polysaccharide contents but not from the poly(L-lysine) homopolymer. Polysaccharide moieties on the surface of the nanoparticles were found to interact specifically with a particular lectin as verified by the aggregation assay. The polynucleotide adsorption capacity of the nanoparticles was increased with increasing polysaccharide contents in the graft copolymers, suggesting that the adsorption conformation of poly(L-lysine) moiety in the graft copolymer on the nanoparticle surface is different from that in poly(L-lysine) homopolymer. Moreover, the nanoparticles from the graft copolymer exhibited resistance against self aggregation and nonspecific adsorption of serum proteins, presumably due to the polymer brush effect and/or exclusion effect from the polysaccharide graft chains. These results suggest that the nanoparticles prepared from poly(L-lysine) graft-polysaccharide copolymer and poly(D,L-lactic acid) can serve as a good DNA carrier in vivo. PMID- 9327140 TI - Synthesis of green fluorescent protein-ricin and monitoring of its intracellular trafficking. AB - We performed genetic engineering to fuse enhanced green fluorescent protein (EGFP) to the N terminus of RTA, expressed the fusion protein in Escherichia coli, purified and reassociated EGFP-RTA with plant RTB, and purified EGFP-ricin by size exclusion HPLC. The fusion heterodimer was able to bind galactosides, intoxicate cells, and show strong fluorescence. Mammalian cells incubated with EGFP-ricin showed strong cell surface fluorescence at 4 degrees C and, on incubation at 37 degrees C, distributed initially to endosomes and then to Golgi vesicles. Variable sensitivity of mammalian cells to ricin and ricin fusion proteins may be due in part to different patterns of intracellular routing. Cells were incubated with ricin or EGFP-ricin, and inhibition of protein synthesis was measured. Human hepatocellular carcinoma Hep3B cells were 10-fold more sensitive to ricin and 85-fold more sensitive to EGFP-ricin than human epidermoid carcinoma KB cells. Epifluorescence microscopy of cells incubated with EGFP-ricin showed greater localization of the fluorescence signal in the Golgi compartments in Hep3B cells than in KB cells. These data support a model requiring a Golgi dependent step in cell intoxication by ricin. The work further identifies the usefulness of green fluorescent protein fusions in the study of retrograde transport of internalized peptides. PMID- 9327141 TI - Functionalized tricarbocyanine dyes as near-infrared fluorescent probes for biomolecules. AB - The syntheses of three novel functionalized tricarbocyanine dyes are described. These dyes containing isothiocyanate and succinimidyl ester functional groups are reactive toward primary amines and can be used as fluorescent probes for biologically pertinent compounds such as amino acids and functionalized dideoxynucleotides. The absorption and fluorescence maxima occur in the near-IR region of the spectrum (770-810 nm). The succinimidyl ester proved to be very sensitive to hydrolysis and was generated in situ to label amino acids. The isothiocyanates were less susceptible to hydrolysis and were conjugated using organic modified [40% (v/v) acetonitrile] buffers to amino acids. A dye with an alkyl isothiocyanate moiety showed conjugation to amino-functionalized dideoxynucleotide triphosphates. PMID- 9327142 TI - Preparation and properties of oligodeoxynucleotides containing 5-iodouracil and 5 bromo- and 5-iodocytosine. AB - The behavior of oligonucleotides containing 5-iodouracil, 5-bromocytidine, and 5 iodocytidine in concentrated ammonia is described. 5-Aminouracil and 5 aminocytidine are obtained as side products when deprotection is performed at 60 degrees C. Small amounts, if any, of side products are obtained when ammonia deprotection is performed at room temperature. The base-pairing properties of these 5-halopyrimidines including triple helix are described. PMID- 9327143 TI - Facile synthesis of a high-affinity ligand for mammalian hepatic lectin containing three terminal N-acetylgalactosamine residues. AB - A simple cluster glycoside containing three residues of N-acetylgalactosamine with proper inter-residual distances can be a high-affinity ligand for asialoglycoprotein receptor of mammalian liver. YEE(ah-GalNAc)3 [Lee, R. T., and Lee, Y. C. (1987) Glycoconjugate J, 4, 317-328] is such a ligand having a Kd in the subnanomolar range, and this high-affinity ligand has been successfully utilized in the delivery of gene to the parenchymal cells of the liver [Merwin, J. R., Noell, G. S., Thomas, W. L., Chiou, H. C., DeRome, M. E., McKee, T. D., Spitalny, G. L., and Findeis, M. A. (1994) Bioconjugate Chem. 5, 612-620; Hangeland, J. J., Levis, J. T., Lee, Y. C., and Ts'o, P. O. P. (1995) Bioconjugate Chem. 6, 695-701]. Reported here is a synthetic procedure for an equally effective, homologous trivalent ligand, YDD(G-ah-GalNAc)3. The advantage offered by this new cluster glycoside is that the synthetic scheme accomplishes purification of reaction intermediates and the product without chromatographic separations. This greatly simplifies the procedure and allows scale-up of the operation at reduced cost of production. PMID- 9327144 TI - Conjugation of unprotected trisuccin, N-[tris[2-[(N hydroxyamino)carbonyl]ethyl]methyl]succinamic acid, to monoclonal antibody CC49 by an improved active ester protocol. AB - For the conjugation of the trihydroxamate bifunctional chelating agent N-[tris[2 [[N-(benzyloxy)amino]-carbonyl]ethyl]methyl]succinamic acid (trisuccin, 1) to antibodies, we originally used the corresponding 2,3,5,6-tetrafluorophenyl active ester followed by the postconjugation removal of the benzyl protecting groups by catalytic hydrogenation. It was of interest to us to design a conjugation protocol capable of incorporating deblocked hydroxamates into peptides and proteins. Reported procedures that were expected to be compatible with the functionalities present in trisuccin were used with no success, as judged by the lack of ability of the products to radiolabel with 188Re. A simple conjugation method was then developed utilizing the o-nitrophenol (ONP) activated ester of the unprotected trisuccin, N-[tris[2-[(N hydroxyamino)carbonyl]ethyl]methyl]succinamic acid, 3, which eliminates the need for the postconjugation deblocking. An assay for indirect estimation of the active ester content, based on the concentration of its decomposition byproduct, ONP-OH, was developed. Comparison of the indirectly estimated concentrations with those obtained directly from purified products showed > 90% accuracy for this assay. This procedure has the advantage of rapidly using the unpurified active ester, eliminating the possibilities of its decomposition through solvolysis or self-condensation by the unprotected hydroxamate functions. A colorimetric assay was developed for estimation of the number of ligands per molecule of protein. This assay and the fact that all conjugates consistently radiolabeled with 188Re show that this procedure conjugated the unprotected hydroxamate ligands to the CC49 monoclonal antibody. These results indicate the potential applicability of this technique to conjugation of unprotected hydroxamate derivatives with other proteins and peptides. PMID- 9327145 TI - Synthesis of 17 beta-hydroxy esters of 4-estren-17 beta-ol-3-one and carbenicillin, ticarcillin, or functionalized oxacillin: potentially useful conjugates for beta-lactamase-based homogeneous immunoassays. AB - On the basis of the large range of kinetic constants of their substrates, beta lactamases seem to be interesting enzymes for the development of homogeneous immunoassays. For this purpose, hapten-penicillin or -cephalosporin conjugates have to be prepared. The aim of this work is to couple the anabolizing steroid nandrolone to several penicillins characterized by extremely low K(m) and kcat values: ticarcillin, carbenicillin, and oxacillin. The easy decarboxylation of derivatives of phenylmalonic acid (carbenicillin) and thienylmalonic acid (ticarcillin) imposes the choice of very mild procedures which have been specifically adapted to each substance investigated. 4-Estren-17 beta-ol-3-one hemiphenylmalonate is conjugated to 6-aminopenicillanic acid after 1,1' carbonyldiimidazole activation, while 4-estren-17 beta-ol-3-one hemi(3 thiophene)malonate is coupled to 6-aminopenicillanic acid after activation using methanesulfonyl chloride. Before conjugation of oxacillin, a carboxylated analogue of its side chain has been prepared. A procedure resorting to tert-butyl ester protection of the carboxyl group present on the isoxazole ring allows the binding of nandrolone to the remaining carboxyl followed, after specific deprotection, by the conjugation to 6-aminopenicillanic acid giving the oxacillin derivative. In this way, conjugates retaining immunological properties of nandrolone and high inhibiting power of beta-lactamases should be obtained. PMID- 9327146 TI - Microdissection-based genetic discovery and analysis applied to cancer progression. PMID- 9327147 TI - Is routine frozen section assessment feasible in the practice environment of the 1990s? PMID- 9327148 TI - Optimizing combination chemotherapy for Kaposi's sarcoma. PMID- 9327149 TI - ras Mutation: does it have clinical import? PMID- 9327150 TI - The role of frozen section analysis of margins during breast conservation surgery. AB - PURPOSE: The role of frozen section analysis during breast conservation surgery is undefined. Assessment of margins using permanent section evaluation is the standard method of ensuring complete tumor excision. If the margin is positive, however, surgical re-excision is necessary to reduce the likelihood of subsequent local recurrence. Therefore, biopsy of the surgical cavity with immediate pathological evaluation during lumpectomy was performed to evaluate the effect on local recurrence, the number of re-excisions, and cosmesis. PATIENTS AND METHODS: One hundred sixty patients underwent attempted lumpectomy with frozen section margin determination. One hundred forty patients were available for long-term follow-up (mean = 57 months, median = 46 months). All patients underwent attempted breast conservation surgery, which consisted of tumorectomy with excision of a greater than 1-cm rim of grossly normal tissue. Tumor margins were obtained by intraoperative biopsy with frozen section analysis of the lumpectomy cavity walls. RESULTS: In 21 patients (15%), frozen section analyses (FSA) revealed positive margins, resulting in immediate re-excision. In seven of these patients (5%), margins were persistently positive, and these patients therefore underwent mastectomy. Fourteen patients were successfully re-excised to a negative margin. The sensitivity and specificity of FSA were 91% and 100%, respectively. Five percent of patients definitively managed by lumpectomy with FSA of margins recurred locally. The mean cosmesis score after radiotherapy was 7.0 out of a possible 10, correlating with a good to excellent result. DISCUSSION: The accuracy of FSA, low recurrence rate, avoidance of reoperation, and good cosmesis indicate that intraoperative frozen section analysis should be adopted as a safe and effective method of margin analysis during breast conservation surgery. PMID- 9327152 TI - Prognostic implications of c-Ki-ras2 mutations in patients with advanced colorectal cancer treated with 5-fluorouracil and interferon: a study of the eastern cooperative oncology group (EST 2292) AB - PURPOSE: Mutations in c-Ki-ras2 (ras) occur in about 40% of patients with colorectal cancers and occur early in the pathogenesis of this disease. To evaluate the prognostic value of mutations in ras, the Eastern Cooperative Oncology Group (ECOG) conducted a retrospective study (EST 2292) to determine the frequency of mutations in patients with advanced colorectal cancer, and to determine whether ras mutations were associated with altered response to therapy and survival. PATIENTS AND METHODS: Patients were enrolled from four studies: P Z289, an ECOG phase II trial of 5-fluorouracil (5-FU) and interferon (IFN) in patients with advanced colorectal cancer; P-Z991, an ECOG phase I trial of 5-FU and IFN in patients with advanced malignancies; and two trials from the Albert Einstein College of Medicine in patients with advanced colorectal cancer treated with 5-FU and either IFN-alpha or IFN-beta. All patients had advanced colorectal carcinoma and had sufficient histologic material available for analysis for the presence and type of ras, using polymerase chain reaction and dot-blot analysis with sets of probes sufficient to detect all the common mutations of ras at codons 12, 13, and 61. RESULTS: Seventy-two patients were enrolled in this trial. Mutations in ras were detected in 25 (35%), including 17 (23%) in codon 12, four (6%) in codon 13, and four (6%) in codon 61. There was no correlation between the presence of a ras mutation and age, sex, Dukes' stage, histology, or tumor markers. Thirty-one of 72 patients (43%) responded to therapy with 5-FU and IFN, and 10 of 31 responders (32%) and 15 of 41 nonresponders (37%) had mutations in ras. There was no difference in response rates or overall survival between the groups with and without ras mutations. CONCLUSIONS: It is unlikely that ras mutations will have significant prognostic value for either response to therapy or survival in patients with colorectal carcinomas treated with 5-FU and IFN. PMID- 9327153 TI - Tumor control of locally advanced prostate cancer following combined estramustine, vinblastine, and radiation therapy. AB - PURPOSE: A prospective phase II study was carried out to determine whether estramustine phosphate (EMP) plus vinblastine (VBL) in combination with radiotherapy (RT) would improve the control of locally advanced prostate cancer. The rationale for combining EMP plus VBL with RT was based on the clinical and radiobiological data that EMP plus VBL acted as an excellent radiation sensitizer in cultured human prostatic carcinoma cells with the property of tissue selectivity. The combined EMP and VBL were well tolerated in the phase II clinical study of patients with advanced prostate cancer. MATERIALS AND METHODS: Between January 1991 and July 1996, 65 patients, stage T2 (B2) through stage T4 (D1), were entered into the study. Gleason pattern scores ranged from 4 to 10. Pretreatment prostate-specific antigen (PSA) was as follows: < 20 in 21 patients (32%), 20 to 50 in 23 patients (35%), and > 50 in 21 patients (32%). The median age was 70 years (55-83). All patients were treated with megavoltage beam radiation with a total tumor dose of 65 to 70 Gy. Oral EMP 450 mg/m2 daily and VBL 3 mg/m2 weekly were given concomitantly in 46 patients during the 7- to 7 1/2 week course of radiotherapy. RESULTS: All patients showed prompt and complete tumor regression on digital rectal examination at 6 weeks following the completion of treatment. Median follow-up time is 43 months (3-65). PSA fell to an undetectable level by 6 weeks in 56 of 65 patients (86%). For the whole group at 5 years clinical control was 81%, but biochemical control (PSA < 4 ng/mL) was 48%. The likelihood of being free of biochemical relapse at 5 years was a function of initial PSA value (PSA < 20 in 64% of the cases, 21-50 in 60%, and > 50 in 0%). The biochemical-relapse-free survival at 5 years for each stage was T2, 49%; T3, 38%; and T4, 17%. In particular, a group of patients with pretreatment PSA levels of 20 to 50 ng/mL responded quite favorably to the present combined regimen in that only 40% of the patients showed a biochemical failure at 5 years, considering the high level of initial PSA. CONCLUSIONS: The present combined approach is effective in achieving a high rate of tumor control with no disproportionately enhanced side effects. The rapid regression of the tumor nodules and sustained freedom from biochemical relapse suggest excellent long-term tumor control, especially in the group of patients with pretreatment PSA levels of 20 to 50 ng/mL. PMID- 9327151 TI - AIDS Clinical Trials Group Study 094: a phase I/II trial of ABV chemotherapy with zidovudine and recombinant human GM-CSF in AIDS-related Kaposi's sarcoma. AB - PURPOSE: To define the maximum tolerated dose of doxorubicin when combined with fixed doses of bleomycin, vincristine, zidovudine, and recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) in patients with advanced AIDS-related Kaposi's sarcoma. PATIENTS AND METHODS: Twenty male patients were treated with zidovudine at doses of either 100 or 200 mg by mouth every 4 hours, and cytotoxic chemotherapy with bleomycin 10 U/m2 and vincristine 1.4 mg/m2 by vein every 2 weeks. Four successive cohorts received fixed doses of doxorubicin given intravenously every 2 weeks: two cohorts each received 10 mg/m2 (levels 1, 2) or 20 mg/m2 (levels 3, 4). The first cohort received rhGM-CSF at a dose of 10 micrograms/ kg, given subcutaneously on days 2 through 11 (level 1). Due to toxicity, the dose of rhGM-CSF was reduced to 5 micrograms/kg (levels 2, 3) and then to 2.5 micrograms/kg (level 4). RESULTS: The dose-limiting toxicity was severe neutropenia, occurring in 10 patients. Severe neutropenic episodes occurred after a median of three cycles of chemotherapy, with the nadir occurring after 14 days (median). Moderate neutropenia occurred in 14% of all cycles administered. Constitutional toxicities of moderate or greater severity occurred in four patients. Five of 10 patients at a doxorubicin dose of 20 mg/m2 (levels 3 and 4) experienced severe neutropenia. Thus, doxorubicin at 10 mg/m2, with BV (bleomycin, vincristine chemotherapy), zidovudine (100 mg five times daily), and rhGM-CSF (5 micrograms/kg/day), was defined as the maximum tolerated dose. CONCLUSIONS: The maximum tolerated dose of doxorubicin is 10 mg/ m2 every 2 weeks when given in combination with BV chemotherapy, zidovudine, and rhGM-CSF. While the addition of rhGM-CSF at doses of 2.5 to 5 micrograms/kg decreased the duration of neutropenia, it did not prevent the occurrence of severe neutropenia from combined myelotoxic therapy. PMID- 9327154 TI - A dose-seeking trial of edatrexate in combination with vinblastine, adriamycin, cisplatin, and filgrastim (EVAC/G-CSF) in patients with advanced malignancies: promising antineoplastic activity against non-small cell lung carcinomas. AB - PURPOSE: To determine the maximum tolerated dose, toxicities, and potential antitumor activity of edatrexate (E), an antifolate agent with enhanced in vitro antitumor activity as compared with methotrexate (M), when given in combination with vinblastine, doxorubicin, cisplatin, and filgrastim (G-CSF) to patients with advanced malignancies. PATIENTS AND METHODS: Thirty-seven patients with advanced malignancies were treated with escalating doses of edatrexate in combination with vinblastine (V), doxorubicin (A), cisplatin (C), and filgrastim (EVAC/G-CSF) following three different subsequently developed schedules. Schedule 1 was patterned after the MVAC regimen, a combination chemotherapy program with activity against different epithelial malignancies, and consisted of E, 40 mg/m2/day, days 1/15/22; V, 3 mg/m2/day, days 2/15/22; A, 30 mg/m2/ day, day 2; C, 70 mg/m2/day, day 2; repeated every 28 days. Schedules 2 and 3 were designed to avoid observed dose-limiting toxicity on schedule 1 consisting of transient elevation of serum creatinine levels and delayed myelosuppression. Schedule 2 consisted of E, 40 or 60 mg/ m2/day, days 1 and 15; V, 3 mg/m2/day, days 2 and 15; A, 30 mg/m2/day, day 2; C, 30 mg/m2/day, days 1 and 2; cycled every 28 days. Schedule 3 consisted of E, 60 to 120 mg/m2/day, day 1; V, 3 mg/m2/day, day 2; A, 30 mg/m2/day, day 2; C, 30 mg/m2/day, days 1 and 2; cycled every 21 days. Filgrastim 5 micrograms/kg/day was given to all patients subcutaneously until the absolute neutrophil count was greater than 10,000/microL postnadir. Three patients were treated on schedule 1, 10 on schedule 2 (four at an E dose of 40 mg/m2/day and six at an E dose of 60 mg/m2/day), and 24 on schedule 3 (six at each of the following E dosages: 60, 80, 100, and 120 mg/m2/day). RESULTS: Dose limiting toxicities of grade 3 to 4 leukopenia and transient elevation of serum creatinine values were observed in two of three patients treated on schedule 1. A dose-limiting toxicity of grade 3 to 4 leukopenia was noted in two of six patients treated on schedule 2 at an edatrexate dose of 60 mg/m2/day. Two of six patients treated on schedule 3 at an edatrexate dose of 120 mg/m2/day had a dose limiting toxicity of grade 3 stomatitis (one patient) and grade 3 cytopenia (one patient). Nineteen of 37 patients with evaluable or measurable disease had a response to treatment (response rate 51%, 95% confidence intervals = 35%-67%). Nine of 15 patients with metastatic non-small cell lung cancer responded, including one complete remission (response rate 60%, confidence intervals = 35% 85%). A median survival of 517 days (confidence interval = 163-808 days) and a 1 year survival rate of 60% (confidence interval = 35%-85%) was seen in patients with advanced non-small cell lung cancer. CONCLUSIONS: The maximum tolerated dose and the recommended phase II dose of edatrexate is 100 mg/m2/day when administered as part of the EVAC/G-CSF program following schedule 3. Promising antineoplastic activity against non-small cell lung carcinomas was observed, and a phase II study is planned. PMID- 9327156 TI - Combinatorial chemistry to develop new drugs. PMID- 9327155 TI - Non-Hodgkin's lymphoma of the paranasal sinuses: clinical and pathological features, and response to combined-modality therapy. AB - PURPOSE: Lymphomas of the paranasal sinuses may have poorer prognoses compared with other extranodal lymphomas of the head and neck, and are not well defined as a particular clinicopathologic entity. The outcome of combined-modality therapy and central nervous system (CNS) prophylaxis has not been fully determined. PATIENTS AND METHODS: We retrospectively reviewed our experience with 16 consecutive, carefully defined patients, all treated with both chemotherapy and radiotherapy. RESULTS: There were 11 men and five women, mean age 52. All presented with local symptoms; 13 had stage I or II disease. Thirteen had diffuse large cell lymphoma, two diffuse mixed, and one small noncleaved. Phenotyping revealed 10 B-cell, four T-cell, and two T or natural killer (NK). Most received CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) chemotherapy; the order of chemotherapy and radiotherapy varied. Twelve received CNS prophylaxis. Of 12 complete responses, six relapsed, all at distant sites, and two died during initial therapy. Five-year survival was 29%, and median survival 18 months. Four of 10 B-lineage patients were relapse-free at 4 years; all six T- or T/NK-lineage patients relapsed or were dead within 6 months. Tumors of T or NK lineage often expressed CD56 and showed evidence of Epstein-Barr viral infection; otherwise, pathological features were not predictive of lineage or outcome. Neither age nor lactate dehydrogenase predicted prognosis. No complete responder recurred in the CNS as site of first relapse. CONCLUSION: Despite localized stage at presentation, sinus lymphoma is an aggressive disease, characterized by distant relapse and early mortality. Combined-modality therapy with CNS prophylaxis improves outcome compared with radiotherapy alone; however, prognosis remains poor. Patients with T-lineage disease appear to have a particularly bad outcome. Autologous bone marrow transplantation should be evaluated as first-line therapy for those at high risk of relapse. PMID- 9327179 TI - Therapy of congestive heart failure in elderly persons. AB - LVEF should be measured in all elderly persons with CHF Underlying causes and precipitating causes of CHF should be treated. Persons with CHF associated with abnormal LVEF should be treated with a low sodium diet, diuretics, and ACE inhibitors. If CHF persists, digoxin should be added. If CHF still persists, isosorbide dinitrate plus hydralazine should be added. If CHF still persists, a beta blocker should also be added. However, calcium channel blockers should not be used. Persons with CHF associated with normal LVEF should be treated with a low sodium diet, diuretics, and ACE inhibitors. If CHF persists, a beta blocker, isosorbide dinitrate plus hydralazine, or a calcium channel blocker should be added to the therapeutic regimen. If sinus rhythm is present, digoxin should not be used. Persons with CHF and abnormal or normal LVEF unable to tolerate ACE inhibitors should be treated with losartan. PMID- 9327180 TI - Initial therapy of advanced prostate cancer. PMID- 9327181 TI - Office management of bronchopulmonary dysplasia. PMID- 9327182 TI - Anorexia nervosa. PMID- 9327183 TI - Malabsorption: a clinical update. PMID- 9327184 TI - Abdominal pain & vomiting in infants & children: imaging evaluation. PMID- 9327185 TI - Diabetics not receiving recommended preventive care. PMID- 9327186 TI - Siemens introduces new diffusion MR imaging tool: a technological advance in early acute stroke diagnosis. PMID- 9327187 TI - UCSF scientists use latest techniques to study biological basis of drug addiction at new center. PMID- 9327188 TI - Knee injuries can take female athletes out of the game. PMID- 9327189 TI - The influence of dosage form on aspirin kinetics: implications for acute cardiovascular use. AB - In this study, the pharmacokinetics of several formulations of aspirin were examined: soluble aspirin, mouth-dispersible aspirin, plain aspirin and enteric coated aspirin granules. Blood samples were taken at frequent intervals for 24 hours after single dosing in 12 healthy volunteers and Tmax, Cmax and t1/2 measured. Cmax was significantly higher for soluble aspirin than for the other formulations and the t1/2 was shorter. The results show the rapid absorption of aspirin from a soluble formulation compared with that from plain aspirin or enteric-coated aspirin granules. Recommendations to treat patients suspected of having a heart attack as soon as possible with aspirin are now widely accepted and the present study would suggest that soluble aspirin should be the aspirin of choice in this situation. PMID- 9327191 TI - A comparison of zopiclone and propiomazine as hypnotics in outpatients: a multicentre, double-blind, randomized, parallel-group comparison of zopiclone and propiomazine in insomniacs. AB - In a double-blind, parallel-group study of 135 patients with a mean age of 60 years, zopiclone 5 mg was compared with propiomazine 25 mg. The patients rated their sleep in a diary. There were statistically significant differences in favour of zopiclone for nine out of 13 variables measuring subjective sleep quality and quantity. Concerning side-effects, bad taste was reported more frequently in the zopiclone group and restless legs in the propiomazine group. PMID- 9327190 TI - A comparison of etodolac (Ultradol) with acetaminophen plus codeine (Tylenol #3) in controlling post-surgical pain in vasectomy patients. AB - The efficacy and safety of etodolac (Ultradol) and acetaminophen plus codeine [A + C (Tylenol #3)] in controlling post-surgical pain were compared in an open label, randomized, parallel-group outpatient study. Patients who were voluntarily having a vasectomy performed for sterilization were assigned to receive either etodolac 200 mg (20 patients) or A + C (20 patients). All medication was taken as required for up to 7 days. Efficacy assessments were made at 1, 6 and 24 hours after surgery and included pain measurement (Likert Visual Analogue scale), patient and physician global assessments and time to analgesic relief. Safety assessments were made throughout the study and included vital signs and adverse event monitoring. Results of the study indicated that patients taking etodolac were more likely to say they could return to work 24 hours after their vasectomy (p = 0.04). There were no other statistically significant differences between the two groups of patients. The results from this study indicate that etodolac and A + C are equally efficacious and well-tolerated for the control of post-vasectomy pain and that patients may observe an increased benefit with etodolac by being able to return to work sooner. PMID- 9327192 TI - Controlled clinical trial on the efficacy and safety of oral sulodexide in patients with peripheral occlusive arterial disease. AB - One hundred and seven adult outpatients with Leriche stage II peripheral occlusive arterial disease took part in this open, controlled trial. Patients were randomly treated over a six-month period either with sulodexide capsules containing 250 lipoproteinlipase releasing units (LRU, two capsules twice daily for 176 days on average: 56 patients), or with pentoxifylline 400 mg tablets (one tablet three times a day for 180 days on average: 51 patients). The incidences of diabetes, hyperlipoproteinaemias, smoking habit and other risk factors were the same in the two groups. The drugs' efficacies were evaluated by monitoring, at the start of treatment and every month during it, the Winsor Index and the walking distance, both prior to (initial claudication distance-IDC) and after (absolute claudication distance-ACD) the symptom's onset. Compliance with treatment and occurrence of adverse events were constantly monitored; systemic tolerability was evaluated through the use of routine haematological and haematochemical tests. Both treatments brought about a progressive increase in the claudication-free walking distance, statistically significant versus baseline from the second month (ACD, sulodexide group) and third month (ACD and ICD, pentoxifilline and sulodexide groups). At the end of treatment, the absolute increase of ACD was significantly greater in sulodexide-treated patients (p < 0.01) with respect to the pentoxifylline-treated group. In both groups the Doppler test evidenced a good improvement in local arterial haemodynamics. In the sulodexide group, 3.6% of patients developed nausea, dyspepsia and other minor gastrointestinal phenomena. In the pentoxifylline group 17.6% of patients complained of gastroenteric disorders (nausea, vomiting, dyspepsia), or of headache and dizziness. In one patient of this latter group insomnia was also present. Systemic tolerance of both drugs was consistently good. PMID- 9327193 TI - An open, parallel group comparison of quinapril and captopril, when added to diuretic therapy, in the treatment of elderly patients with heart failure. AB - This study aimed to compare the efficacy, tolerability and first-dose blood pressure response of once-daily quinapril and twice-daily captopril when added to diuretic therapy in elderly patients with heart failure. The study was performed at a single centre as an open randomised parallel-group study, patients being selected for inclusion from the outpatient population. Following a starting dose of either 2.5 mg once-daily quinapril, or 6.25 mg twice-daily captopril, patients were reviewed at two-weekly intervals, and following clinical assessment a decision was made either to titrate up to the next medication stage or to enter the patient into the 16-week maintenance phase. Efficacy was assessed using a six minute walking test, the New York Heart Association (NYHA) class, a functional lifescale (FLS) questionnaire and the cardiothoracic ratio (CTR)-at study entry and at the end of the maintenance phase. Blood pressure was measured for 5 h post first-dose of medication. Sixty-one patients were randomised to treatment: 30 to quinapril and 31 to captopril. Following withdrawals, data from 36 patients (20 on quinapril, 16 on captopril) were available for analysis. The distance walked during the six-minute walking test improved in both groups; the difference between the treatment groups was not statistically significant. There were no significant changes in the FLS or CTR. An analysis of change in the NYHA status from study entry to study end showed a statistically significant difference between the two groups (p = 0.02) in favour of quinapril. Five patients in each group experienced hypotension during the 5 h following the first dose of medication. This study has shown heart failure to be as well controlled by once daily quinapril as by twice-daily captopril, with comparable effects on first dose blood-pressure response. PMID- 9327194 TI - Rifaximin, a non-absorbable rifamycin, for the treatment of hepatic encephalopathy. A double-blind, randomised trial. AB - The aim of this study was to evaluate the efficacy and tolerability of rifaximin, a non-absorbable intestinal antibiotic, in comparison to neomycin in the short- and long-term treatment of hepatic encephalopathy (HE). Forty-nine patients with a definite diagnosis of cirrhosis were included in this double-blind, randomised, controlled trial. Patients were randomly assigned to one of the following treatments: (1) rifaximin 400 mg three times daily; (2) neomycin 1 g three times daily. Both drugs were administrated orally as tablets during 14 consecutive days each month, for a period of six months. The neuropsychiatric signs and blood ammonia levels were examined before starting the treatment, and every 30 days, until the final assessment. In all patients a progressive and important reduction in HE grade was observed, and no statistically significant difference between the two treatments was detected. In both groups the disturbances in speech, memory, behaviour and mood, gait, asterixis, writing, and serial subtraction of 7 s and five-pointed star tests all showed the highest proportion of improvement. During the study blood ammonia levels decreased in both the rifaximin and in the neomycin groups, and again no statistically significant difference was found between groups. Our findings confirm, therefore, the usefulness of rifaximin in the treatment of HE, supporting its use as a first-choice antibiotic, particularly in patients intolerant to neomycin or with impaired renal function. PMID- 9327195 TI - Is cholesterol the major lipoprotein risk factor in coronary heart disease?--a Franco-Scottish overview. AB - There has been much debate over the past three decades concerning the role of hyperlipidaemia in coronary heart disease (CHD) and the efficacy of reducing plasma lipids levels. Although reduction in plasma cholesterol has been associated with a favourable effect on both primary and secondary CHD, there is a growing feeling that cholesterol may not be the only significant lipoprotein risk factor to be involved. Only relatively recently has the true role of triglycerides become apparent. Studies have indicated that the greatest reduction in CHD with some treatments has been found in those patients in whom high triglyceride levels accompany hypercholesterolaemia. In particular, in younger patients who have suffered a myocardial infarction, hypertriglyceridaemia is more common than hypercholesterolaemia. Nevertheless, recent large studies have shown that reduction of low-density lipoprotein (LDL) is beneficial, even in post infarction patients with a relatively normal total cholesterol level. Furthermore, studies with fibrates and with HMG Co-A reductase inhibitors have indicated that progression of atheromatous lesions can be halted and in may cases there is evidence of regression. Continuing research on the pathophysiology of atherosclerosis, including the role of macrophages and thrombotic involvement, will further define the role of hypolipidaemics in the prevention and management of coronary heart disease. PMID- 9327196 TI - Efficacy and tolerability of azelastine nasal spray in the treatment of allergic rhinitis: large scale experience in community practice. AB - Two Spanish prospective monitoring studies evaluated efficacy and tolerability of azelastine nasal spray containing azelastine hydrochloride for allergic rhinitis. Both studies were conducted by community practitioners over two weeks (Study I) or one month (Study II). The numbers of patients recruited were 3680 (I) and 4002 (II). Of these, 56.1% (I) and 51.7% (II) had been previously treated with oral antihistamines with/without other medications. Patients rated the severity of 10 symptoms of allergic rhinitis as absent, mild, moderate or severe. Azelastine nasal spray was generally administered at a dose of one spray puff (0.14 mg) per nostril twice daily. Follow-up was after 14 days (I) or 31 days (II), when symptoms were rated and patients questioned about treatment. Assessment was by a sum score for all 10 symptoms. A symptom sum score of 16-20 occurred in 21.1% (I) and 13.7% (II) of patients before treatment and only 0.8% (I) and 0.6% (II) after treatment. A symptom sum score of 11-15 occurred in 35.9% (I) and 30.5% (II) of patients before treatment and only 2.6% (I) and 2.8% (II) after treatment. Overall, 92.3% (I) and 90.7% (II) of patients were completely free of adverse events, 7.0% (I) and 8.8% (II) experienced one and 0.7% (I) and 0.6% (II) two adverse events. The number of doctors who rated efficacy as either very good or good was 89.4% (I) and 84.6% (II). General tolerance was rated as good or very good by 97.5% (I) and 97.3% (II), and local tolerance by 93.1% (I) and 91.5% (II) of physicians, respectively. Overall, azelastine nasal spray was highly effective and very well tolerated in normal clinical practice. PMID- 9327197 TI - HIV antibodies: further questions and a plea for clarification. AB - The existence of specific antibody/protein reactions is the crucial assumption underlying proof of HIV isolation, proof of HIV infection and the causative role of HIV in AIDS. However, since 1. antibodies which react with the 'HIV' proteins arise following allogenic stimuli in non-HIV-infected animals and humans, as well as in mice and humans with autoimmune disorders; antibodies to antigens from both mycobacteria and yeasts cross-react with HIV env and gag proteins; 2. individuals belonging to the AIDS risk groups are subjected to allogenic stimuli and have high levels of autoimmune antibodies, while the vast majority of patients in the AIDS risk groups are infected with either or both mycobacteria or yeasts; the evidence for the existence of HIV and its putative role in AIDS must be reappraised. PMID- 9327198 TI - Molecular basis of inherited disorders of renal solute transport. PMID- 9327199 TI - Sodium taste. AB - The regulation of sodium metabolism is achieved by the adaptive sodium-saving capacities in epithelia and by hormones acting within the brain to modulate salt appetite. Taste, especially in rodents, has a sodium-specific component that provides a guiding function for salt intake. The taste responsiveness is not invariable, however, and may be modified in the case of sodium need. This review discusses emerging functional aspects of the peripheral and central branch of the salt sensory pathway. PMID- 9327200 TI - Renal osmoregulatory transport of compatible organic osmolytes. AB - Cells in renal medullas are exposed to a high concentration of salt during antidiuresis. They adapt in part by accumulating myo-inositol, glycine betaine, taurine, and other amino acids by transporting them from the interstitial fluid. This transport is osmotically regulated by changes in transcription of the transporters and by post-translational modifications. Most of the original studies were in vitro, but these processes are increasingly being examined in vivo. PMID- 9327201 TI - Na,K-ATPase. AB - Na,K-ATPase is the driving force for renal Na+ reabsorption and is thus critically implicated in the control of extracellular volume and blood pressure. This review focuses on most recent advances in the elucidation of intrinsic structural features that are important for Na,K-ATPase function and in the identification of regulatory mechanisms that are implicated in the modulation of Na,K-ATPase activity in the kidney under physiological or pathophysiological conditions. PMID- 9327202 TI - Sodium-hydrogen exchange in renal epithelia: mechanisms of acute regulation. AB - Five isoforms of sodium-hydrogen exchangers are present in the kidney. Specific isoforms serve specialized functions and are strategically located in polarized distributions in different nephron segments. This review updates understanding of the renal distribution and highlights some of the recent advances in understanding the mechanisms of acute regulation of sodium-hydrogen exchangers in the kidney. PMID- 9327203 TI - Regulation of ion transport by protein-protein interaction domains. AB - Protein-protein interaction via specific modular domains has been implicated in the regulation of many signalling pathways. Recently, such modules, in particular WW, Src homology-3 and PDZ domains, have been shown to regulate localization, clustering and function of several ion channels and transporters, including the epithelial Na+ channel, K+ channel and ionotropic neurotransmitter receptors. PMID- 9327204 TI - Sorting and trafficking of ion transport proteins in polarized epithelial cells. AB - Transport proteins are targeted to specific plasmalemmal domains in polarized epithelial cells. The molecular signals that govern these sorting events are just beginning to be elucidated. In many cases, the cell surface delivery of transport proteins is subjected to tight regulation. Several different mechanisms appear to participate in these trafficking processes. PMID- 9327205 TI - Vasopressin V2-receptor antagonists: panaceas for hyponatremia? AB - The current treatment of hyponatremia is unsatisfactory and can be associated with significant morbidity. Vasopressin is inappropriately elevated in the majority of patients with hyponatremia and causes free water retention by stimulating V2-receptors in the collecting ducts. Recently, orally active, nonpeptide, selective vasopressin V2-receptor antagonists have been characterized and offer an exciting prospect for the treatment for hyponatremia. V2-receptor antagonists are effective aquaretic agents, that are capable of increasing free water clearance and plasma sodium and might be useful in the treatment of hyponatremia caused by syndrome of inappropriate secretion of antidiuretic hormone, heart failure, cirrhosis, and nephrotic syndrome. The rationale for their use and evidence from animal and human studies are discussed. PMID- 9327206 TI - Enhancing endogenous effects of natriuretic peptides: inhibitors of neutral endopeptidase (EC.3.4.24.11) and phosphodiesterase. AB - Because diuretic drugs remain the main treatment for disorders of sodium and water metabolism, the quest for improved diuretic and natriuretic agents continues in the hope of achieving fewer side effects and a more rational basis in pathophysiology. One aim has been to enhance endogenous diuretic and natriuretic activity by selective manipulation of atrial natriuretic peptide and related compounds. The first approach has been to inhibit degradation of these peptides using inhibitors of their main catabolic enzyme, neutral endopeptidase, and to offset any antagonistic effect of the renin-angiotensin system by combination with an angiotensin-converting enzyme inhibitor. The second and more recent approach has been to inhibit breakdown of the second messenger of atrial natriuretic peptide, cGMP, using phosphodiesterase inhibitors. As yet, neutral endopeptidase inhibition has not advanced successfully beyond animal experimentation and phosphodiesterase inhibition is still in its infancy. Both strategies suffer from the problem that, on the one hand, neutral endopeptidase metabolizes a variety of bioactive peptides, including endothelin, and it is not possible to develop inhibitors that will be selective for a given peptide; whereas, on the other hand, there are several phosphodiesterase isoforms metabolizing cGMP and cAMP, both second messengers for many different bioactive compounds, and selective inhibitors are still under development. PMID- 9327207 TI - The rise and fall of atrial natriuretic peptide for acute renal failure. AB - Atrial natriuretic peptide can increase glomerular filtration rate and filtration fraction and can promote natriuresis, effects that would logically seem to improve renal function after acute tubular necrosis from ischemic or toxic injury. Early human trials suggested a beneficial effect of atrial natriuretic peptide on creatinine clearance, and a reduction in the need for dialysis in treated patients. However, randomized, placebo-controlled trials have failed to show a clinically relevant benefit on survival, dialysis-free survival, or renal function in patients treated with this agent. PMID- 9327208 TI - Dietary salt restriction: benefits for cardiovascular disease and beyond. AB - The evidence linking salt intake to high blood pressure has become stronger. At the same time, there is increasing evidence that salt has other adverse effects independent of its effects on blood pressure. PMID- 9327209 TI - Hyperhomocysteinemia: detection, risk assessment, and treatment. AB - Homocysteine is formed by the demethylation of methionine in the course of its normal metabolism. Hyperhomocysteinemia is an independent risk factor for vascular disease. It develops most commonly from folate deficiency, genetic abnormalities, and chronic renal failure. Current models favor direct angiotoxicity involving endothelial and vascular smooth muscle cells, and impaired thrombolysis. Folic acid reduces hyperhomocysteinemia and thus provides an opportunity for risk-factor modification. PMID- 9327210 TI - Angiotensin-converting enzyme inhibitor therapy for non-diabetic progressive renal disease. AB - Chronic renal disease evolves to end-stage renal failure through events, including enhanced intraglomerular pressure and plasma protein ultrafiltration, mediated at least in part by angiotensin II. Angiotensin-converting enzyme inhibitors reduce intracapillary pressure and ameliorate glomerular size selective function, which may account for their antiproteinuric effect and renoprotective potential. Thus, the Ramipril Efficacy in Nephropathy study found a significant correlation between enhanced urinary protein excretion and faster disease progression in non-diabetic patients with proteinuric chronic renal disease. In proteinuric non-diabetic renal disease at comparable levels of blood pressure control, angiotensin-converting enzyme inhibitors reduce proteinuria and slow disease progression to end-stage renal failure safely and more effectively than non-angiotensin-converting enzyme therapy. On the contrary, most non proteinuric chronic renal diseases progress slowly and do not benefit specifically from angiotensin-converting enzyme inhibition therapy. PMID- 9327211 TI - Gene therapy for the treatment of renal disease: prospects for the future. AB - Human gene therapy has been progressing at a remarkable rate, but information from basic research on gene therapy is still generally lacking. Gene transfer to the kidney has been attempted using various strategies, yet we still have no satisfactory and reliable gene transfer technique to the kidney. More effective and more selective gene transfer techniques are required for gene therapy, as well as for use as a research tool, to improve understanding of the pathophysiology of renal diseases. PMID- 9327212 TI - Treatment of hypertension in the elderly. AB - Hypertension is present in over 50% of elderly patients and constitutes a major risk factor for cardiovascular morbidity and mortality. This paper reviews the rationale for treating hypertension in the elderly, discusses the choice of antihypertensive therapy and optimal target blood pressure, and summarizes ongoing clinical trials. The major questions that remain to be answered are the optimal level of blood pressure reduction in the elderly and the long-term efficacy and safety of newer antihypertensive agents compared with diuretics and beta-blockers. PMID- 9327213 TI - Molecular cell biology and physiology of solute transport. PMID- 9327214 TI - Pharmacology and therapeutics. PMID- 9327215 TI - Lymphoma. PMID- 9327216 TI - Recent progress in lymphoma classification. AB - In the Revised European-American list of lymphoid neoplasms recently proposed by the hematopathologists of the International Lymphoma Study Group, the disease entities currently recognized in the literature among the B-cell and T/natural killer (NK)-cell lymphomas and leukemias and variants of Hodgkin's disease are collected and defined. This proposal is here to stay. The main challenge for the future is to present the information in this list in a form that facilitates the understanding, learning, and teaching of these disorders. A meaningful classification for clinical use has already been built on the foundation provided by this list, through the agreement of an international group of clinicians. Newer data confirm the clinical validity of many categories in the list. Others suggest the need for additions or possible changes. To be discussed, among the latter, are: a need for a better definition of the new category of lymphoplasmacytic lymphoma; the variants of mantle-cell lymphoma and their correlation with genomic abnormalities; the need for a clarification of the relationship among the variants listed as "marginal zone B-cell" lymphomas; the subclassification of large B-cell lymphomas; and the coming of age of the NK-cell lymphomas. PMID- 9327217 TI - Recent advances in lymphoma biology. AB - This review highlights three topics in which major contributions to lymphoma biology have recently been achieved. First, the use of single-cell polymerase chain reaction analysis has allowed for a better understanding of the origin of several entities, eg, Hodgkin's, Burkitt's, and marginal zone B-cell lymphomas. Second, the studies of mechanisms regulating apoptosis and survival of the neoplastic clones have opened a whole area of research with promising expectations for new therapeutical approaches. Finally, the recent cloning of genes involved in several chromosomal translocations is likely to shed more light on the molecular mechanisms responsible for the neoplastic transformation. The survey presents the main results obtained in these three areas and critically discusses them in the light of the current debate. PMID- 9327218 TI - Treatment of aggressive histology lymphoma. AB - The recent phase III intergroup trial showed no improvement in outcome with the use of second- and third-generation regimens for primary treatment of aggressive lymphomas. Efforts are now underway to significantly intensify therapies for patients with high-risk aggressive lymphoma by utilizing hematopoietic growth factors or autologous peripheral blood stem cells. Preliminary results show minimal or no benefit to intensification of standard regimens using growth factor support alone. However, high-risk patients who achieve a complete remission with standard therapy appear to benefit from consolidative bone marrow transplantation. Results of a novel, dose-intense regimen, which administers several non-cross-resistant drugs as high-dose single agents in rapid succession, may provide a promising alternative for primary therapy of high-risk aggressive lymphoma. New regimens incorporating high-dose cytarabine for patients with poor prognosis, small noncleaved-cell lymphoma seem to improve survival. The ability to effectively tailor therapy for subgroups of patients with aggressive lymphoma depends on the continued identification of clinical and biologic prognostic factors to compliment the International Prognostic Factors Index. PMID- 9327219 TI - Treatment of non-Hodgkin's lymphoma with high-dose therapy and hematopoietic stem cell support. AB - Standard therapies for advanced low-grade lymphomas are not likely to provide a cure, prompting the use of more aggressive approaches. Patients with low-grade lymphoma who receive high-dose therapy with stem cell support appear to have a prolonged disease-free survival, although the benefit to overall survival remains unproven. Patients with chemotherapy-sensitive intermediate- or high-grade relapsed disease have improved survival with high-dose therapy, and those with high-risk disease may benefit from early consolidation while in first remission. Significant questions remain in terms of the proper timing of high-dose therapy, appropriate stratification by risk factors, the value of purging, the role of radiotherapy after transplantation, and the most appropriate source of stem cells for transplantation. PMID- 9327220 TI - Late sequelae of treatment of Hodgkin's disease. AB - Twenty-three years after it was observed that successful treatment of Hodgkin's disease was associated with risk of a second malignancy related to treatment, the literature is maturing. Early reports of leukemia risk rising and falling between 5 and 10 years after treatment continue to be supported. The real problem is the steady emergence of solid tumors in radiation treatment fields, particularly breast cancer and in females treated between the ages of 10 and 25. Several reports this year again emphasize this point. One avenue to pursue to avoid this complication is to use combination chemotherapy alone, especially in young women. The other relates to one observation made this year that low doses of radiation may not be as carcinogenic as full doses. These data suggest that chemotherapy and low-dose radiotherapy in early stage Hodgkin's disease need our attention in rigorously designed clinical trials. PMID- 9327221 TI - Treatment of epidemic (AIDS-related) Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) is the most common tumor seen in patients with HIV-1 infection. KS causes significant morbidity and mortality through involvement of the skin and visceral organs. The optimal treatment for KS depends on the extent of the disease and immunologic status. However, with knowledge gained on the pathogenesis of disease, newer therapies and compounds are being developed. Early disease patients are best treated with either local therapy or agents that have low toxicity and can be delivered long term. Advanced disease, such as in patients with widespread mucocutaneous disease, lymphedema, and visceral disease, are treated most effectively with cytotoxic agents such as liposomal anthracyclines, vinca alkaloids, or paclitaxel. Future treatment developments are focusing on the role of effective anti-HIV therapy and anti-human herpesvirus (HHV)-8 therapy in an effort to interfere with key steps in the etiology of KS to control the disease. Secondly, agents that focus on the interruption of autocrine and paracrine growth factors such as vascular endothelial cell growth factor and basic fibroblast growth factor, interleukin-6, and interleukin-8 are of therapeutic interest. Some of these compounds currently under evaluation include antiangiogenesis inhibitors and retinoids. PMID- 9327222 TI - Human herpesvirus-8. AB - Our current understanding of Kaposi's sarcoma (KS) has evolved from initial descriptions of this "idiopathic sarcoma" to a progressively detailed characterization of its probable infectious origin. This article traces the explosive discovery of human herpesvirus-8 (HHV-8) and its etiologic associations with KS, Castleman's disease, and primary effusion lymphomas. A framework for understanding how viral infection leads to development of these unusual disease is presented. The possible role of HHV-8 in tumorigenesis, through viral manipulation of signal transduction pathways and cell cycle control, is discussed. PMID- 9327223 TI - HIV-associated lymphomas. AB - Intermediate and high-grade non-Hodgkin's lymphomas (NHL) with a B-cell phenotype are AIDS-defining illnesses. The incidence of systemic NHL increased greatly and primary central nervous system NHL increased even more in the HIV-infected population. Further increases in frequency are anticipated as HIV-infected individuals survive longer in an immunosuppressed state with improved antiretroviral treatment and treatment of opportunistic infections. Unusual types of NHL and manifestations of Hodgkin's disease are seen in HIV-infected individuals also. The pathologic and clinical features of the HIV-associated lymphomas and treatment approaches and results are the subjects of this review. Other articles in this issue discuss epidemiology and pathogenesis. PMID- 9327224 TI - The use of new antiretroviral therapy in combination with chemotherapy. AB - With rapid scientific advances in HIV pathogenesis and antiretroviral therapy, the oncologist is faced with new challenges in the treatment of patients with HIV infection and malignancy. HIV infection, by causing immune dysregulation and cytokine production, may alter the natural history of certain neoplasms. Treatment of cancer with chemotherapy can also affect the course of HIV disease. The use of combination antiretroviral agents, particularly the protease inhibitors, can maximally suppress replication of HIV. Improved clinical outcome is associated with more profound decreases in plasma HIV RNA, although development of resistant strains may hinder sustained antiretroviral activity. Antiretroviral therapy should be continued during chemotherapy with the goal of achieving plasma HIV-RNA levels below 500 copies/mL. Modifications in the regimen should be done when intolerable toxicity occurs or if viral load is increased. Although information regarding the use of newer antiretroviral agents with chemotherapy is limited, the use of several reverse transcriptase inhibitors has been well tolerated in patients with AIDS-related malignancies such as Kaposi's sarcoma and lymphoma. PMID- 9327225 TI - Biology of ovarian cancer. AB - Although ovarian carcinomas are thought to arise from the ovarian surface mesothelial layer, the possibility that they develop from mullerian remnants within paraovarian tissues merits further consideration. Molecular genetic studies suggest that ovarian cystadenomas, low malignant potential tumors, and carcinomas are not part of a disease continuum but do represent separate disease entities. Recent advances in our understanding of the molecular genetic changes associated with ovarian epithelial tumor development can be summarized in a working genetic model for ovarian tumorigenesis, which can provide a framework for further studies. Certain molecular changes, such as telomerase expression and alterations in DNA methylation levels, are associated with both tumors of low malignant potential and carcinomas but not with cystadenomas. Mutations in the p53 gene and the development of multiple losses of heterozygosity are specific for the malignant phenotype. The nature of the specific chromosomes affected by the latter losses in a given tumor dictates its biologic aggressiveness. PMID- 9327226 TI - Advances in gynecologic radiation oncology. AB - Radiation therapy is an important modality in the curative and palliative management of patients with gynecologic malignancies. However, the specific roles of radiation therapy continue to evolve in terms of specific indications, volumes treated, techniques, and integration with other modalities. Retrospective assessments of outcome and prospective clinical trials are necessary to answer questions, generate additional hypotheses, and guide further refinements in the application of radiation therapy. This article focuses on recent literature and ongoing clinical trials in this disease group. PMID- 9327227 TI - Decisions about prostate cancer screening in managed care. AB - This review contrasts a traditional biomedical model with an outcomes model and illustrates the distinction in approaches with regards to prostate cancer. The traditional biomedical model emphasizes diagnosis and treatment of medical conditions. The principal data are diagnosis and disease status. In contrast, an outcomes model emphasizes life expectancy and health-related quality of life. According to the traditional model, screening for prostate cancer is encouraged and focus is on the number of cases diagnosed and the success of treatment. Treatment success is measured by recurrence rates or disease-free survival; however, among all men with the potential for a diagnosis of prostate cancer, most will die of other causes. Treatment may have significant consequences in terms of quality of life without clear evidence that it improves survival. The outcomes model argues that decisions for screening and treatment of prostate cancer must be shared between an informed patient and his physician. PMID- 9327228 TI - Experimental models of gene-environment interaction for cancer chemoprevention studies. AB - The recent development of mouse strains with cancer-related genes overexpressed or inactivated has provided investigators with new models for testing chemoprevention strategies to offset specific genetic susceptibilities to cancer. This review focuses on the three genetically altered mouse models that have been the most widely used in chemoprevention studies: Min mice, which carry a mutation in the adenomatous polyposis coli (APC) gene; APC-knockout mice; and p53-knockout mice. Studies with the Min and APC-knockout mice provide the strongest evidence to date that the enzyme cyclooxygenase-2 plays a major role in colon carcinogenesis, and that nonsteroidal anti-inflammatory drugs that target cyclooxygenase-2 have great potential as colon cancer chemopreventive agents. In addition, chemoprevention studies in mice deficient of the p53 tumor-suppressor gene, the most commonly altered gene in human cancer, suggest that the increased susceptibility to cancer resulting from the loss of p53 function may be offset by preventive approaches. Other recently developed transgenic and knockout models of potential interest for chemoprevention studies will also be discussed. PMID- 9327229 TI - Prevention of liver cancer. AB - Hepatocellular carcinoma (HCC) is among the most prevalent and deadly cancers worldwide. Elevated incidence of HCC is strongly associated with hepatitis B virus infection and dietary exposure to hepatotoxic contaminants. Vaccination programs against viral hepatitis hold hope for the eventual reduction of HCC incidence, and screening of cirrhotic patients may provide one mechanism for improving prognosis of HCC patients. Additional efforts have focused on preventing or retarding liver cancer development in populations at risk for HCC mortality. This review highlights recent clinical trials aimed at exploring the efficacy and practicality of chemoprevention efforts in these cohorts. Preclinical findings that provide insight into mechanisms of hepatocarcinogenesis and point to potential strategies for intervention also are discussed. Finally, novel molecular mechanisms contributing to the known ability of chemopreventive agents to inhibit liver cancer in experimental models are explored. PMID- 9327231 TI - Cancer in AIDS. PMID- 9327230 TI - Lymphoma. PMID- 9327232 TI - Gynecologic cancer. PMID- 9327233 TI - Prevention. PMID- 9327235 TI - EEG activation in 1-month-old infants of depressed mothers. AB - Previous research has documented differences in the pattern of EEG activation between 3-month-old infants of depressed mothers and infants of nondepressed mothers. In the present study, EEG was recorded in even younger 1-month-old infants of depressed and nondepressed mothers. The infants of depressed mothers exhibited greater relative right frontal EEG asymmetry (due to reduced left frontal activation), and this pattern at 1 month was significantly related to 3 month EEG asymmetry. Right frontal EEG asymmetry was also related to more frequent negative facial expressions (sad and pre-cry faces) during the Brazelton exam. Finally, the infants of depressed mothers showed more indeterminate sleep, were less active, and cried less than infants of nondepressed mothers. PMID- 9327236 TI - Maternal directive and facilitative interaction styles: associations with language and cognitive development of low risk and high risk toddlers. AB - The purpose of this study was to examine the relation of maternal interaction styles to the development of a sample of 56 toddlers (19 low risk, 37 high risk) seen at 12 and 24 months of age. At 12 months, videotapes of mother-child interaction were coded for directiveness, sensitivity, and elaborativeness. At 12 and 24 months, cognitive and language measures were collected. A directive maternal style was negatively correlated with sensitivity and elaborativeness, whereas sensitive and elaborative ratings were positively correlated, suggesting a facilitative style. Regression models significantly predicted receptive language and cognitive development at 24 months but not expressive language. Maternal directiveness at 12 months was negatively relative to later receptive language skills, whereas elaborativeness at 12 months was positively predictive of later cognitive development. Child status variables and maternal interactional styles contributed about equally to the prediction of later cognitive and language outcomes. PMID- 9327234 TI - Relationship of prenatal cocaine exposure and maternal postpartum psychological distress to child developmental outcome. AB - Maternal cocaine use during pregnancy can affect the infant directly through toxic effects or indirectly through cocaine's influence on maternal psychological status. We followed 160 cocaine exposed and 56 nonexposed infants and their mothers identified at birth through interview and/or urine screen. Although cocaine exposure defined the groups, infant exposure to alcohol, marijuana, and tobacco was allowed to vary. Infants were 99% African American and poor. All mothers completed the Brief Symptom Inventory (BSI) and infants were given the Bayley Scales of Mental (MDI) and Motor (PDI) Development at a mean corrected age of 17 +/- 8 months. Both MDIs (94 +/- 17 vs. 103 +/- 16) and PDIs (101 +/- 16 vs. 108 +/- 12) were lower for cocaine exposed infants. Psychological distress was greater in cocaine using mothers. Hierarchical multiple regression was used to assess the relative effects of gestational age, maternal psychological distress, and cocaine and polydrug exposure on infant outcomes. Both psychological distress and cocaine and alcohol exposure predicted lower MDIs after controlling for prematurity. Neither psychological distress nor alcohol exposure predicted motor outcome, while cocaine had a significant effect. Tobacco and marijuana exposure were unrelated to outcome. These findings provide further support for direct effects of cocaine and alcohol on infant development as well as highlight the need for studies to document maternal psychological factors, which may increase child risk for poorer outcomes. PMID- 9327237 TI - The Preschool Assessment of Attachment: construct validity in a sample of depressed and nondepressed families. AB - Construct validity of the newly developed Preschool Assessment of Attachment (PAA) was examined in a sample of depressed and nondepressed mothers and their preschoolers, focusing on attachment related differences in children's general caregiving environments, maternal psychosocial functioning, and child behavior during interactions with mother. Mothers of secure children were more emotionally and verbally responsive to their children than were mothers of insecure children, and secure children were emotionally more positive to their mothers than were insecure children. Mothers of secure children also reported higher levels of social supports than did mothers of insecure children. Finally, dyads with children who lacked unitary, coherent attachment strategies (i.e., anxious depressed, defended/coercive, and insecure other) showed the worst functioning in all domains relative to all other attachment groups. Similar but slightly less robust findings were obtained with socioeconomic variables statistically controlled. These results lend support to the PAA as a valid system for the conceptualization and measurement of quality of attachment among preschoolers. Future research applications with the PAA are discussed. PMID- 9327238 TI - Mother-toddler interaction patterns associated with maternal depression. AB - Interactive coordination was observed in laboratory play interactions of pairs of 29 clinically depressed and 14 nondepressed mothers and their 13-29-month-old children (M = 18.9 months). Nondepressed mothers and their children displayed more interactive coordination than depressed-mother dyads (p < .001). Depressed mothers were less likely to repair interrupted interactions, and their toddlers were less likely to maintain interactions than nondepressed controls. Toddlers matched their nondepressed but not their depressed mothers negative behavior rates. Results suggested that early interventions focus on training mothers to attend to maintain, and repair mother-child interactions to more closely approximate normal levels of interactive coordination. PMID- 9327239 TI - Preventive intervention as means of clarifying direction of effects in socialization: anxious-withdrawn preschoolers case. AB - An indicated preventive intervention research program integrating attachment, attributional, and behaviorist perspectives was conducted to test the hypothesis that parent-child relationship disturbances directly effect the child's adjustment to the preschool. Anxious-withdrawn preschool children and their mothers were divided equally into treatment and control groups, and assessed on maternal self-report of parenting stress, behavioral ratings of mother-child interaction, and teacher ratings of the children in the preschool classroom. Results showed significant changes in the treatment of group: mothers in the treatment group moderated their level of control to a more appropriate, less intrusive level, while children in the treatment group showed an increase in cooperation and enthusiasm during a problem solving task with mother. Teacher rated social competence and anxious-withdrawn behavior indicated improvement, although only the former was significant. The demonstration of effects of this home intervention for the mother on the child's behavior in the preschool confirm the transactional model underlying this study and demonstrate the utility of a parent-child interaction training component for the prevention of behavioral emotional problems in young children. PMID- 9327240 TI - Follow-up study of young stress-affected and stress-resilient urban children. AB - Reports follow-up study of 181 young highly stressed urban children, classified as stress-resilient (SR) and stress-affected (SA) 1 1/2-2 years earlier. At follow-up (T2), children were retested on five initial (T1) test measures: self rated adjustment, perceived competence, social problem solving, realistic control attributions, and empathy; parents and teachers did new child adjustment ratings, and parents participated in a phone interview focusing on the T1-T2 interval. Child test and adjustment measures and parent interview responses at T2 sensitively differentiated children classified as SR and SA at T1. Test and interview variables used at T1 and T2 correlated moderately across time periods. At T2, four child test indicators (i.e., rule conformity, global self-worth, social problem solving, and realistic control attributions) and four parent interview variables (positive future expectations for the child, absence of predelinquency indicators, good parent mental health in the past year, and adaptive parent coping strategies) sensitively differentiated children classified as SR and SA at T1. No relationship was found between family stress experienced in the T1-T2 interval and changes in children's adjustment during that period. PMID- 9327241 TI - Emotion recognition in autism: verbal and nonverbal information. AB - This study examined the roles of verbal and nonverbal sources of information in the ability of persons with and without autism to recognize emotion. Child, adolescent, and young adult participants in four groups [Lower Functioning Autism (LFA) (n = 17), High Functioning Autism (HFA) (n = 18), Lower Functioning Comparison (LFC) (n = 18), and High Functioning Comparison (HFC) (n = 23)] identified emotions shown (happy, angry, sad, surprised, or neutral) in video clips of individuals expressing emotion verbally, nonverbally, or both. Verbal expressions of emotion were either Explicit, Implicit, or Neutral, whereas nonverbal expressions were Animate or Flat (3 x 2). Pairwise ANCOVAs indicated no group differences between HFA and HFC groups or between the LFA and LFC groups, and indicated instead group differences between higher and lower functioning persons. With groups collapsed into High Functioning (HF) and Lower Functioning (LF), significant group differences were found. Performance of LF individuals suggested they had difficulty inferring how a person felt based on what the person said, if the emotion was not explicitly named. Performance of HF individuals suggested they relied more on nonverbal than on verbal information to determine a speaker's emotion, except where the emotion was explicitly named. Results suggested that persons with autistic spectrum disorders can use affective information from multiple sources in much the same ways as persons of comparable developmental level without autism. PMID- 9327242 TI - A review of the in vitro activity of meropenem and comparative antimicrobial agents tested against 30,254 aerobic and anaerobic pathogens isolated world wide. AB - The in vitro activity of meropenem (formerly SM-7738), a new carbapenem, was compared with that of imipenem and five other broad-spectrum antimicrobials (ceftazidime, cefotaxime, piperacillin, piperacillin/tazobactam, and ciprofloxacin) against 30,254 clinically significant pathogens isolated in nine countries worldwide. Overall, the carbapenems, meropenem and imipenem, were the most active drugs. Meropenem was four- to 64-fold more active than imipenem against Gram-negative bacteria, including the Enterobacteriaceae, Pseudomonas aeruginosa, Burkholderia cepacia, Haemophilus influenzae, and Neisseria meningitidis. Meropenem was also quite active against ceftazidime-resistant strains of Enterobacteriaceae, inhibiting 87.5 to 100% at < or = 4 micrograms/ml. In contrast, imipenem was four- to eight-fold more active than meropenem against Gram-positive species, including methicillin-susceptible strains of Staphylococcus aureus and Staphylococcus epidermidis, Streptococcus pneumoniae, and Enterococcus faecalis. Among the anaerobes, strains resistant to meropenem or imipenem were encountered very rarely. These extensive data provide additional in vitro support for the clinical use of meropenem as a broad spectrum antimicrobial agent active against key pathogenic species of bacteria. PMID- 9327243 TI - Antimicrobial resistance and serotypes of Shigella isolates in Kigali, Rwanda (1983 to 1993): increasing frequency of multiple resistance. AB - The serotype distribution and susceptibility to nine antibiotics was determined for 2491 Shigella isolates cultured in the medical laboratory of the Centre Hospitalier de Kigali, Rwanda, during 1983 to 1993. Overall, Shigella flexneri was the most frequent species, ranking before Shigella sonnei, Shigella boydii, and Shigella dysenteriae. However, the relative frequency of the different Shigella spp. showed an important variability over time. S flexneri increased from 40% in 1983 to 68% of the isolates in 1993 whereas S. dysenteriae Type 1 decreased gradually from 30 to 0.5% of the isolates in 1992. After the outbreak of severe civil unrest, which caused the displacement of many people to the capital, a new epidemic of dysentery started in the Kigali area and S. dysenteriae Type 1 accounted again for 24% of the isolates in 1993. In 1983, resistance to tetracycline, streptomycin, and sulfonamides was common among the endemic Shigella spp. Resistance to chloramphenicol was observed in 17% (30/182) of the isolates. Only 10% were resistant to ampicillin and an equal proportion to trimethoprim, whereas 5% of the isolates showed resistance to both products. By 1993, 66% (195/295) of the isolates were resistant to chloramphenicol (for comparison with 1983, p < 0.001), 70% (207/295) to ampicillin (p < 0.001), 67% to trimethoprim (p < 0.001), and 58% had combined resistance to the latter two drugs (p < 0.001). Resistance patterns differed strongly by species, S. flexneri being more frequently resistant than S. sonnei. In 1983, all S. dysenteriae Type 1 isolates were resistant to ampicillin, chloramphenicol, tetracycline, and sulfonamides. Trimethoprim resistance increased from 31% (25/80) in 1983 to 96% (26/27) of the isolates in 1986 (p < 0.001). After the introduction of nalidixic acid as an alternative for trimethoprim-sulfamethoxazole, trimethoprim resistance decreased to 87%, during 1987 to 1992, and subsequently to 68% of the isolates in 1993. However, 20% of the isolates became resistant to nalidixic acid in 1993. Ampicillin and trimethoprim-sulfamethoxazole are no longer useful for the empirical treatment of shigellosis in Rwanda. PMID- 9327244 TI - Comparison of polymerase chain reaction and pulsed-field gel electrophoresis for the epidemiological typing of Alcaligenes xylosoxidans subsp. xylosoxidans in a burn unit. AB - Eighteen isolates of Alcaligenes xylosoxidans subsp. xylosoxidans were collected from clinical specimens of 15 patients in a burn unit and a plastic surgery ward over a 16-month period. Pulsed-field gel electrophoresis and polymerase chain reaction (PCR) were compared for the epidemiologic typing of these 18 isolates and fifteen epidemiologically unrelated strains. These 18 isolates demonstrated an identical fingerprint pattern and were easily distinguished from the 15 epidemiologically unrelated strains by pulsed-field gel electrophoresis typing and both enterobacterial repetitive intergenic concensus and repetitive extragenic palindrome-primed PCR fingerprinting. We conclude that pulsed-field gel electrophoresis analysis of XbaI-digested genomic DNA is a highly discriminatory and reproducible method for epidemiological typing of A. xylosoxidans subsp. xylosoxidans isolates. However, poor resolution due to frequent cutting in the smaller fragments (< 145.5 Kb) may lead to difficulty in interpretation. PCR is a rapid and highly discriminatory, but less reproducible, technique with occasional loss of major bands. The fingerprints produced by repetitive extragenic palindrome primed PCR had more intense bands and were easier to read than those produced by enterobacterial repetitive intergenic concensus-primed PCR in this study. PMID- 9327245 TI - In vitro antifungal activity of BMS-181184 against systemic isolates of Candida, Cryptococcus, and Blastomyces species. AB - BMS-181184 is a water-soluble derivative of the pradimicin group of antifungal compounds. We determined the in vitro activities of BMS-181184 and comparator agents amphotericin B, 5-fluorocytosine, fluconazole, and ketoconazole against 184 systemic fungal isolates collected at the Health Sciences Centre in Winnipeg, Canada, between 1987 and 1995. BMS-181184 demonstrated MICs of between 1 and 8 micrograms/mL for all Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei, Candida lusitaniae, and Cryptococcus neoformans isolates tested. BMS-181184 was less active against Candida parapsilosis (MIC90 = 16 micrograms/mL) and Blastomyces dermatitidis (MIC90 = 32 micrograms/mL). Isolates of Candida species with fluconazole MICs of > or = 16 micrograms/mL and those with fluconazole MICs of < or = 8 micrograms/mL demonstrated similar BMS-181184 sensitivities. PMID- 9327246 TI - Activity of selected beta-lactams, ciprofloxacin, and amikacin against different Acinetobacter baumannii biotypes from Chilean hospitals. AB - The activity of some third generation cephalosporins, aztreonam, imipenem, ciprofloxacin, and amikacin against isolates of Acinetobacter baumannii of various biotypes has been studied. The isolates, independently of the biotype, exhibited a broad multiresistance against cephalosporins. Ceftazidime was the most active and cefoperazone the least active compound. Aztreonam also showed low activity and no imipenem-resistant strains were found. Ciprofloxacin and amikacin were somewhat more active than cephalosporins, but resistant isolates were also frequent. Isolates of Biotypes 9 and 8 exhibited broader multiresistance than those of Biotype 6 and "other." PMID- 9327247 TI - In vitro activities of antimicrobial agents, alone and in combinations, against Burkholderia cepacia isolated from blood. AB - Burkholderia cepacia is a widespread, environmental gram-negative bacillus that is associated with nosocomial infections. This bacterium is considered to be an important pathogen in immunocompromised patients and is inherently resistant to multiple antimicrobial agents. To compare the activity of different antimicrobial agents and the potential of combinations against invasive strains of B. cepacia, we collected 36 isolates of B. cepacia from blood cultures and checked their susceptibilities to 13 antimicrobials by broth microdilution method. Most strains tested were susceptible to minocycline (94.4%), ceftazidime (86.1%), ciprofloxacin (83.3%), and trimethoprim-sulfamethoxazole (83.3%). All strains were resistant to aminoglycosides, and only some strains were susceptible to imipenem (16.7%), aztreonam (19.4%), moxalactam (25.0%), piperacillin (25.0%), and carbenicillin (47.2%). The effects of combinations of ceftazidime with amikacin, ceftazidime with ciprofloxacin, and ciprofloxacin with amikacin were assayed by checkerboard titration method. Synergistic effect was found in 28 out of 36 tested strains (77.8%), when ceftazidime was combined with amikacin, in 25 out of 36 strains (69.4%) when ceftazidime was combined with ciprofloxacin, and in only 8 out of 36 strains (22.2%) when ciprofloxacin was combined with amikacin. PMID- 9327248 TI - Antimicrobial activity of trovafloxacin tested against ciprofloxacin-susceptible and -resistant Neisseria gonorrhoeae. Interpretive criteria and comparisons with Etest results. AB - Trovafloxacin, a new fluorinated naphthyridine, has enhanced activity against Gram-positive cocci, while retaining an excellent spectrum against Gram-negative pathogens. It has been used successfully in clinical trials for therapy of gonorrhea, and this investigation proposes in vitro susceptibility testing criteria for trovafloxacin. A total of 150 Neisseria gonorrhoeae clinical isolates (50 resistant to ciprofloxacin; MICs > or = 0.12 microgram/mL) were tested by methods recommended by the National Committee for Clinical Laboratory Standards (NCCLS) and the Etest (AB BIODISK, Solna, Sweden). Trovafloxacin was very active against gonococci (MIC90, 0.008 to 0.015 microgram/mL), but was generally eightfold less potent versus ciprofloxacin-resistant strains. Etest results correlated well (r = 0.96; 98% of MICs +/- one log2 dilution) compared to the reference agar dilution test. Reference agar dilution and Etest MICs were compared to disk-diffusion test zones (10-micrograms trovafloxacin disk), and excellent categorical agreement (89.4 to 99.3%) was achieved without significant false-susceptible or -resistant error (< or = 1.3%). Tentative breakpoints were suggested initially to outline the ciprofloxacin-susceptible and trovafloxacin susceptible as susceptible (MIC, < or = 0.015 microgram/mL; zones > or = 47 mm), and strains with various well-characterized mutations of the gyr A and par C genes as either intermediate or resistant to trovafloxacin. When the results of clinical studies treating ciprofloxacin-resistant N. gonorrhoeae with trovafloxacin become available, the alternative breakpoints could be utilized for resistant MIC breakpoints of > or = 0.06 microgram/mL or > or = 0.5 microgram/mL. Trovafloxacin was stable in supplemented GC agar for 21 days stored at refrigerated temperatures. These in vitro results indicate that trovafloxacin should be a very acceptable agent for therapy of gonorrhea and other common sexually transmitted pathogens. PMID- 9327249 TI - Serum inhibitory titers and serum bactericidal titers for human subjects receiving multiple doses of the antibacterial oxazolidinones eperezolid and linezolid. AB - In Phase I trials subjects received multiple doses of eperezolid (PNU-100592; formerly U-100592) and linezolid (PNU-100766; formerly U-100766), and steady state samples were drawn at the projected peak and trough timepoints. Serum inhibitory titer and serum bactericidal titer values were determined using single strains of Staphylococcus aureus, Enterococcus faecalis, and Streptococcus pneumoniae. Serum inhibitory titer values generally correlated with drug concentration in serum and inherent organism susceptibility. Against S. aureus and E. faecalis sera from patients dosed with either drug were generally inhibitory at the peak timepoint, but at trough only linezolid exhibited a persistent effect. No bactericidal activity was seen for either drug against S. aureus or E. faecalis. The sera from patients dosed with either drug exhibited inhibition of S. pneumoniae at peak and trough. Bactericidal activity was seen against S. pneumoniae for both drugs at peak time and at trough for many of the sera for patients on the higher dose regimens. The results demonstrated that the sera from most human subjects dosed with eperezolid or linezolid were inhibitory to S. aureus and E. faecalis and S. pneumoniae and that many of the samples exhibited bactericidal activity for S. pneumoniae. PMID- 9327250 TI - Vertebral osteomyelitis: nontypeable beta-lactamase-negative Haemophilus influenzae in an adult: case report. AB - Adult pyogenic vertebral osteomyelitis due to Haemophilus influenzae is exceedingly rare. After a search of the literature, we deemed our case to be the seventh case of H. influenzae pyogenic osteomyelitis. Vertebral osteomyelitis in itself is a rarity. The most common organisms associated with vertebral osteomyelitis are Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Of the six previously reported cases of adult pyogenic vertebral osteomyelitis due to H. influenzae, four of the six cases were caused by Type B H. influenzae, one case was attributed to Type C, and the other strain was not typed. PMID- 9327251 TI - Inducible amp C beta-lactamase producing gram-negative bacilli from blood stream infections: frequency, antimicrobial susceptibility, and molecular epidemiology in a national surveillance program (SCOPE). AB - A surveillance study of nosocomial blood stream infections [Surveillance and Control of Pathogens of Epidemiologic Importance (SCOPE)] was conducted during a 14-month period in 1995 to 1996 in approximately 50 American medical centers. Among the 4725 blood stream infections, the etiologic agent was Enterobacter spp. in 230, Citrobacter freundii in 24, and Serratia marcescens in 65. The vast majority of these isolates (89%) had been sent to the University of Iowa including 198 Enterobacter spp. (46 Enterobacter aerogenes, 141 Enterobacter cloacae, 11 other Enterobacter spp.), 23 C. freundii, and 62 S. marcescens. Because these species are capable of producing Amp C beta-lactamase, we examined their susceptibility to 12 broad-spectrum antimicrobial agents. The frequency of resistance to ceftazidime and the molecular epidemiology of ceftazidime-resistant strains was also examined. Among the Enterobacter spp. and C. freundii isolates, resistance to third generation cephalosporins (ceftazidime, ceftriaxone) and broad-spectrum semisynthetic penicillins (piperacillin), with or without an enzyme inhibitor (piperacillin/tazobactam), was high, e.g., 35 to 50%. The S. marcescens isolates were quite susceptible to all agents tested. Both imipenem and cefepime were active against virtually all isolates tested including 84 stably derepressed Amp C-producing ceftazidime-resistant strains of Enterobacter spp. and C. freundii. The overall rank order of activity for the six best agents against these Amp C-producing strains was: imipenem (100% susceptible) > amikacin = cefepime (98.6%) > ciprofloxacin = gentamicin = ofloxacin (93.6 to 94.0%). Molecular typing studies of ceftazidime-resistant E. cloacae using an automated ribotyping system, as well as pulsed-field gel electrophoresis, indicated that although clonal spread of a single strain occurred in some of the medical centers, most of the episodes of bacteremia were caused by patient-unique strains. Control of these resistant organisms will require attention to microbiologic recognition of phenotypes, to infection control practices, and to limiting the overuse of certain extended spectrum beta-lactams. PMID- 9327252 TI - Susceptibility of ninety-eight clinical isolates of Legionella to macrolides and quinolones using the Etest. AB - Isolates of Legionella from 98 patients with Legionnaires' disease hospitalized in Columbus, Ohio, USA between 1991 through 1995 were tested for antimicrobial susceptibility to macrolides and quinolones using the Etest. Most (87%) isolates were Legionella pneumophila serogroup 1. All isolates tested remain susceptible to erythromycin, azithromycin, clarithromycin, ciprofloxacin, ofloxacin, and levofloxacin. In vitro susceptibility testing of Legionella to representative macrolides and quinolones should be considered to detect the emergence of resistant isolates. PMID- 9327253 TI - The morbidity of insomnia uncomplicated by psychiatric disorders. AB - The morbidity of sleep problems has been well documented; however, they are frequently associated with and are symptomatic of several psychiatric disorders. It is unclear how much of the morbidity can be accounted for by the associated psychiatric and substance abuse disorders and medical problems, and how much by the sleep problems per se. Sleep problems may also be an early sign of a psychiatric problem. This paper reports data from an epidemiologic community survey of over 10,000 adults living in three U.S. communities. A structured diagnostic assessment of psychiatric disorders as well as assessment of the presence of insomnia not due to medical conditions, medication, drug or alcohol abuse, and a 1-year follow-up were completed. Persons with insomnia in the past year without any psychiatric disorders ever (uncomplicated insomnia); with a psychiatric disorder in the past year (complicated insomnia); and with neither insomnia nor psychiatric disorders ever were compared on treatment utilization and the first onset of a psychiatric disorder in the subsequent year. Eight percent of those with uncomplicated as compared with 14.9% with complicated insomnia and 2.5% with neither had sought treatment from the general medical sector for emotional problems in the 6 months prior to the interview. The rates of treatment sought from the psychiatric specialty sector were 3.8%, 9.4%, and 1.2%, respectively. These differences were significant after controlling for sociodemographic characteristics and were sustained when the persons were interviewed 1 year later. Uncomplicated insomnia was also associated with an increase in risk for first onset of major depression, panic disorder, and alcohol abuse over the following year. Insomnia, even in the absence of psychiatric disorders, is associated with increased use of general medical and mental health treatment for emotional problems and for the subsequent first onset in the following year of some psychiatric disorders. Early diagnosis and treatment of uncomplicated insomnia may be useful. PMID- 9327254 TI - Treatment of somatization in primary care. AB - A large proportion of patients present to primary care with chronic, stress related symptoms having no organic cause. Biomedical treatment of these patients is usually ineffective and expensive. A 6-week behavioral medicine intervention designed to provide adjunctive treatment to primary care was evaluated in a randomized, controlled study. Thirty-eight individuals receiving treatment and 44 waiting for treatment completed the SCL-90-R at times corresponding to 1 week before (time 1) and 1 week after the course (time 2). The treatment group was then followed up at 6 months. After correction for initial levels, the treatment group reported significantly less somatization, anxiety, and depression than did the wait-list group at time 2. Within the treatment group, decreases in somatization, anxiety, and depression were statistically significant and were maintained 6 months later. Within the wait-list group, distress remained unchanged. A review of relevant literature reveals that a general behavioral medicine course such as the one studied here has an important adjunctive role in primary care, since 1) subsyndromal psychological distress is common in primary care; 2) physicians are reluctant to address psychosocial issues; 3) negative mood is associated with poor health; 4) negative mood is associated with high, inappropriate medical utilization; and 5) negative mood is associated with help seeking behavior. PMID- 9327256 TI - Medical evaluation of persons with mental retardation referred for psychiatric assessment. AB - Many people with developmental disabilities and "challenging behaviors" present to primary care physicians, internists, or general psychiatrists for assessment and treatment. These clinicians seek to provide the comprehensive biopsychosocial assessment necessary for successful treatment, but may encounter interference from funding agencies. Epidemiologic data on medical comorbidity in persons with developmental disabilities with a primarily "behavioral" presentation may assist in facilitating these assessments. A total of 1135 people with mental retardation referred for mental health assessment were medically evaluated according to a two step protocol which included a screening evaluation of all persons and expanded testing, depending on clinical status. The workup was considered complete when the person with either improving clinically or had a specific terminal diagnosis and was as comfortable as possible. Medical comorbidity was about double that of people referred for mental health assessment who do not have mental retardation. Common conditions presented in unusual ways, and less frequent conditions presented more often. The cost of the medical assessments was promptly recovered in a variety of savings to systems. Comprehensive medical assessment discloses increased medical comorbidity in persons with mental retardation referred for psychiatric evaluation. Comprehensive treatment based on the assessment findings appears to be associated with better clinical outcomes and cost savings to systems. PMID- 9327255 TI - Medical training in psychiatry residency. A proposed curriculum. AB - During the coming decades, psychiatrists will be asked to participate to a greater extent in the physical evaluation and treatment of patients with behavioral or emotional problems. Despite the high frequency with which psychiatric symptoms are caused or exacerbated by organic disease, psychiatrists have been reluctant, and in some ways, even discouraged to include physical assessments. Psychoanalysis and concerns about boundary issues have influenced psychiatrists to cede physical assessment and physical illness to other physicians. To help overcome these barriers to improved care of psychiatric patients, a curriculum is proposed for psychiatry residents. It will allow them to better use their medical backgrounds while increasing their contributions as mental health specialists. PMID- 9327257 TI - Neuropsychiatric aspects in Munchausen's syndrome. AB - The case of a 27-year-old woman with Munchausen's syndrome is presented, in whom a history of cerebral palsy and pathological findings from electroencephalogram, cranial computed tomography, and neuropsychological assessment were prominent. It is argued that neuropsychiatric aspects may play a role in the development of Munchausen's syndrome in a subgroup of patients. PMID- 9327258 TI - Use of high dose benzodiazepines in alcohol and sedative withdrawal delirium. AB - The authors describe two patients who required massive doses of benzodiazepines to treat complicated alcohol and sedative withdrawal delirium. Some of the factors that contribute to difficulties in management are discussed. Finally, we describe the advantages and disadvantages of high dose pharmacologic management and controversies regarding psychopharmacologic management of such complex patients. PMID- 9327259 TI - The connection between the psychological condition of breast cancer patients and survival: a follow-up after eight years. PMID- 9327260 TI - Trinucleotide GAA repeat expansions in seven French Friedreich ataxia families. AB - We collected 7 Friedreich ataxia (FRDA) pedigrees from France. All cases but one family were homozygous for an unstable GAA trinucleotide expansion in the first intron of the frataxin gene. In this peculiar pedigree absence of the GAA expansion supports the notion of possible genetic heterogeneity of FRDA. PMID- 9327261 TI - Severe mental retardation with marfanoid habitus in a young Lebanese male. A diagnostic challenge. AB - A severely mentally retarded 20-year-old male adult with a marfanoid habitus reported since birth and dysmorphic features is described. A younger brother presented the same appearance at birth but died at 2 months of age following a cardiopathy. Despite some clinical differences, his features are very close to the Lujan-Fryns syndrome and to two unrelated girls described by de Die-Smulders et al. (1). This case shows the challenge that clinicians could have while investigating patients with mental retardation and marfanoid habitus for diagnosis and genetic counseling. PMID- 9327262 TI - Monozygotic twins discordant for the oculo-oto-radial syndrome (IVIC syndrome). AB - Monozygotic twins are rarely completely identical despite their origin from a single zygote. The twinning process itself, or the very early intrauterine environment must provide the clues to explain this developmental dilemma. We present here discordant monozygotic twin girls, one of whom was diagnosed having IVIC (Oculo-oto-radial) syndrome on the basis of hand abnormalities and hearing loss but her twin sister has only strabismus. The family has at least 7 apparently and 2 possible affected members (gene carriers) over four generations, the majority being only midly affected. The 2 girls, whose monozygosity has been proved using independent DNA loci, show marked variability in expression, showing that for this gene modification of expression must be epigenetic or environmental rather than genetic. PMID- 9327263 TI - How to counsel in osteopathia striata with cranial sclerosis. AB - Osteopathia with cranial sclerosis (OS-CS) is an autosomal dominant condition, which is characterized by typical radiological changes of the skull and the long bones, in association with a wide variety of clinical symptoms. A family is reported with at least three affected individuals. One of them, a young women and her husband asked for a preconceptional advice, but the highly variable expressivity, as documented by this family, made it very difficult to counsel them properly. PMID- 9327264 TI - Pyknodysostosis: hemangioma of the skull as a new finding. AB - Pyknodysostosis is a rare sclerosing bone dysplasia syndrome with autosomal recessive inheritance. Here, we report a case of pyknodysostosis, with characteristic physical and radiological findings, but also with a hemangioma of the skull, as a non reported finding sofar. PMID- 9327265 TI - On the nosology of the craniodigital syndromes: report of a family and review of the literature. AB - During a systematic survey for genetic causes of mental retardation in schools for adolescents with learning problems we had the occasion to examine a 16-year old moderately mentally retarded boy with facial dysmorphism, short stature, relative microcephaly, complete cutaneous syndactyly of fingers III/IV and of toes II/III. Partial clinical manifestations (low to subnormal intelligence and syndactyly) were present in his mother and sister. We discuss the nosology and differential diagnosis of the craniodigital syndromes. PMID- 9327266 TI - An exonic polymorphism (381A/G) in the choroideremia gene. AB - By using the single strand conformational analysis (SSCA) to search for point mutations in the choroideremia gene, we have identified a previously undescribed polymorphism within exon 5a (381A/G). We have studied the frequency of this polymorphism in a population from Southern France. The sequence variation creates a new restriction site for HhaI, allowing a convenient DNA-based genetic counseling in families in which the causal disease mutation is unknown. PMID- 9327267 TI - Binder syndrome in a mother and her son. AB - Binder syndrome or maxillonasal dysplasia is characterized by maxillary hypoplasia and a flat, vertical nose. Inheritance is uncertain. We report on a mother and her son with Binder syndrome. The proband was the last child of a kinship of seven children. Three sisters, three brothers and the parents were normal. The proband had a maxillary hypoplasia and a flat, vertical nose. The columella was short, the nostrils had a triangular shape and the upper lip was convex with an acute nasolabial angle. There were no dental abnormalities apart from those secondary to malocclusion. Height and intelligence were normal. After an uncomplicated pregnancy, the proband delivered a boy who had low nasal bridge, hypertelorism, maxillary hypoplasia and a small nose with a broad columella. Although the majority of cases of maxillonasal dysplasia are sporadic, familial occurrence has been reported by a number of authors. In six pedigrees the recurrence was in second or third degree relatives. Recurrence in sibs with unaffected parents has been observed seven times and an affected parent and child has been rarely reported. The Binder phenotype may be heterogeneous. The pattern of abnormalities seen in this condition does not represent a causally defined entity. PMID- 9327268 TI - Social, familial and medical aspects of Usher syndrome in Colombia. AB - A psycho-social study was performed in 19 Colombian families with 40 affected individuals with Usher syndrome, identified through our national screening program for this disease in Colombia. The study was aimed to understand their needs, kind of familial inter-relationships, and social and familial implications of the patients' double sensorial limitation, in order to provide enough information to support the importance of early diagnosis, appropriate genetic counseling, and the establishment of adequate educational and rehabilitation programs in Colombia. PMID- 9327270 TI - Pierre-Robin sequence and severe mental retardation with chaotic behaviour associated with a small interstitial deletion in the long arm of chromosome 2 (del(2)(q331q333)). AB - In this report we describe a severely mentally retarded male presenting an interstitial deletion in the long arm of chromosome 2 (del(2)(q331q333)). He presented a Pierre-Robin sequence at birth, and during childhood increasing behaviour problems were noted. PMID- 9327269 TI - Multiple vertebral segmentation defects. Brief report of three patients and nosological considerations. AB - Multiple vertebral segmentation defects i.e. multiple malformations of vertebrae and ribs are characterized by short neck, scoliosis, short trunk and deformity of the ribcage. There are three major subtypes; Jarcho-Levin syndrome, spondylothoracic dysostosis and spondylocostal dysostosis, with different inheritance patterns, survival rates and associated malformations. We describe three cases of multiple vertebral segmentation defects, two with familial spondylothoracic dysostosis and one with sporadic spondylothoracic dysostosis, and anomalies i.e. super-numerary breast, clubfeet deformity, myelomeningocele, intradural lipoma, and Arnold-Chiari malformation. PMID- 9327271 TI - An adaptive training method for optimal interpolative neural nets. AB - In contrast to conventional multilayered feedforward networks which are typically trained by iterative gradient search methods, an optimal interpolative (OI) net can be trained by a noniterative least squares algorithm called RLS-OI. The basic idea of RLS-OI is to use a subset of the training set, whose inputs are called subprototypes, to constrain the OI net solution. A subset of these subprototypes, called prototypes, is then chosen as the parameter vectors of the activation functions of the OI net to satisfy the subprototype constraints in the least squares (LS) sense. By dynamically increasing the numbers of subprototypes and prototypes, RLS-OI evolves the OI net from scratch to the extent sufficient to solve a given classification problem. To improve the performance of RLS-OI, this paper addresses two important problems in OI net training: the selection of the subprototypes and the selection of the prototypes. By choosing subprototypes from poorly classified regions, this paper proposes a new subprototype selection method which is adaptive to the changing classification performance of the growing OI net. This paper also proposes a new prototype selection criterion to reduce the complexity of the OI net. For the same training accuracy, simulation results demonstrate that the proposed approach produces smaller OI net than the RLS-OI algorithm. Experimental results also show that the proposed approach is less sensitive to the variation of the training set than RLS-OI. PMID- 9327272 TI - Classification of user expertise level by neural networks. AB - A neural network approach to low-level user modeling is described, in the context of text editing tasks using the Jove editor. Knowledge of a user's expertise is extracted automatically, based on their interaction with Jove over a two week period. A MLP classifier which uses rprop learning and incorporates output data fuzzification is developed to classify users into one of five expertise levels. Classification into the correct level is achieved in around 80% of the cases, with misclassification being restricted to adjacent classes. The neurofuzzy system is seen to outperform not only the binary classifier of Beale [1989], but also production rule and inductive expert systems developed especially for comparison purposes in this study. PMID- 9327273 TI - Translation-, rotation-, scale-, and distortion-invariant object recognition through self-organization. AB - The task of visual object recognition is often complicated by the fact that a single 3-D object can undergo a number of transformations which can substantially alter its projection onto a 2-D surface, such as the retina. Such transformations include translation of the object in the visual field, changes in the size of the object, its orientation in the 2-D plane and the viewing perspective. For a general pattern recognition system to detect and recognize and object after such transformations, it must be able to associate widely differing patterns with the same object label. In this paper, a novel self-organizing model, called the Multiple Elastic Modules (MEM), is presented which attempts to solve this problem by searching a multi-dimensional space, where each axis is defined by one of the transformations (e.g. scale, translation, rotation, etc.). A particular object of a specific size, orientation and spatial location is mapped onto a single point in this space. Of course, distortions and minor variations in an object's image will expand this point to a small localized area in this multi-dimensional space. Such a powerful representation scheme comes at a cost of high computational demand due to the combinatorially large search space. The MEM approach to solving this problem efficiently partitions the solution space to search the most promising areas for the correct match. Simulation results are presented on detecting a stick-figure object under translation, distortion, scale, and rotation transformations in a cluttered background. PMID- 9327274 TI - Supervised adaptive Hamming net for classification of multiple-valued patterns. AB - A Supervised Adaptive Hamming Net (SAHN) is introduced for incremental learning of recognition categories in response to arbitrary sequences of multiple-valued or binary-valued input patterns. The binary-valued SAHN derived from the Adaptive Hamming Net (AHN) is functionally equivalent to a simplified ARTMAP, which is specifically designed to establish many-to-one mappings. The generalization to learning multiple-valued input patterns is achieved by incorporating multiple valued logic into the AHN. In this paper, we examine some useful properties of learning in a P-valued SAHN. In particular, an upper bound is derived on the number of epochs required by the P-valued SAHN to learn a list of input-output pairs that is repeatedly presented to the architecture. Furthermore, we connect the P-valued SAHN with the binary-valued SAHN via the thermometer code. PMID- 9327275 TI - Recognition of rotating images using an automatic feature extraction technique and neural networks. AB - This paper presents a new automatic feature extraction technique and a neural network based classification method for recognition of rotating images. The image processing technique extracts global features of an image and converts a large size image into a one-dimensional small vector. A special advantage of the proposed technique is that the extracted features are the same even if the original image is rotated with rotation angles from 5 to 355 or rotated and a little bit distorted. The proposed technique is based on simple co-ordinate geometry, fuzzy sets and neural networks. The proposed approach is very easy in implementation and it has been developed in C++ on a Sun workstation. The experimental results have demonstrated that the proposed approach performs successfully on a variety of small as well as large scale rotated and distorted images. PMID- 9327276 TI - Data mining of inputs: analysing magnitude and functional measures. AB - The problem of data encoding and feature selection for training back-propagation neural networks is well known. The basic principles are to avoid encrypting the underlying structure of the data, and to avoid using irrelevant inputs. This is not easy in the real world, where we often receive data which has been processed by at least one previous user. The data may contain too many instances of some class, and too few instances of other classes. Real data sets often include many irrelevant or redundant input fields. This paper examines the use of weight matrix analysis techniques and functional measures using two real (and hence noisy) data sets. The first part of this paper examines the use of the weight matrix of the trained neural network itself to determine which inputs are significant. A new technique is introduced and compared with two other techniques from the literature. We present our experience and results on some satellite data augmented by a terrain model. The task was to predict the forest supra-type based on the available information. A brute force technique eliminating randomly selected inputs was used to validate our approach. The second part of this paper examines the use of measures to determine the functional contribution of inputs to outputs. Inputs which include minor but unique information to the network are more significant than inputs with higher magnitude contribution but providing redundant information, which is also provided by another input. A comparison is made to sensitivity analysis, where the sensitivity of outputs to input perturbation is used as a measure of the significance of inputs. This paper presents a novel functional analysis of the weight matrix based on a technique developed for determining the behavioral significance of hidden neurons. This is compared with the application of the same technique to the training and test data. Finally, a novel aggregation technique is introduced. PMID- 9327277 TI - On neural blind separation with noise suppression and redundancy reduction. AB - Noise is an unavoidable factor in real sensor signals. We study how additive and convolutive noise can be reduced or even eliminated in the blind source separation (BSS) problem. Particular attention is paid to cases in which the number of sensors is larger than the number of sources. We propose various methods and associated adaptive learning algorithms for such an extended BSS problem. Performance and validity of the proposed approaches are demonstrated by extensive computer simulations. PMID- 9327278 TI - A supervised learning network based on adaptive resonance theory. AB - A neural network architecture, fuzzy ART with logistic discrimination (ART-LD), is introduced as a method of realising the pattern recognition task in a supervised learning manner. The system is formed by the hierarchical organisation of two network modules: a fuzzy ART and a logistic discrimination. The learning consists of two separate stages. Firstly, the fuzzy ART module self-organises the input patterns into category clusters, whose operations are governed by fuzzy set theory and "competitive learning" dynamics that ensure fast and stable learning. Then the outputs, which can be interpreted as fuzzy memberships of an input pattern to the encoded categories, provide the spatial distance information that is generalised by the subsequent logistic discrimination to give the final prediction. Examples are presented, and the generalisation capabilities of ART-LD are demonstrated through two simulated and one real classification problem. PMID- 9327279 TI - Cephalometric parameters affecting severity of anterior open bite. AB - One hundred and four patients with anterior open bite (AOB) were subdivided by statistical means into three distinctly separate groups with AOB of different severity based on the extent of separation of the incisors in the vertical plane. Cephalometric evaluation of all subjects was completed using 24 skeletal and 12 dentoalveolar measurements, and the results were compared between the mild, moderate and severe groups of AOB. The findings suggest that the palatal plane, mandibular occlusal plane, upper anterior dental height and upper posterior dental height correlate significantly with the severity of AOB. In contrast, skeletal jaw relationship in the horizontal plane correlated only weakly with severity. This study further highlighted the steep mandibular occlusal plane as an important indicator of AOB severity in contrast to the maxillary occlusal plane. PMID- 9327280 TI - Internal derangement of the temporomandibular joint: correlation of arthrographic imaging with surgical findings. AB - Sixty-seven patients, who were surgically treated for internal derangement of the temporomandibular joint (TMJ), were retrospectively examined. The patients were evaluated preoperatively by clinical and arthrographic examinations and these results were compared with findings at surgery. Partial or complete dislocation of the disc was detected by arthrographic examination in 65 joints. Actual disc displacement was demonstrated at surgery in 57 TMJs, giving arthrography a positive predictive value of 88% for detecting disc dislocation. Arthrographic diagnosis of disc perforation was unreliable, as both false positive and false negative observations were recorded. Arthrography was found to have a positive predictive value of only 53% for assessing disc perforation. Based upon its proven inaccuracy, invasiveness and discomfort to the patient, it is recommended to replace arthrography by magnetic resonance imaging. PMID- 9327281 TI - Galeal skin island flap in the reconstruction of scalp defects caused by missile injuries. AB - The use of the galeal skin island flap pedicled either on the superficial temporal or occipital vessels for coverage of scalp defects following neurosurgical treatment of cranial missile injuries, is presented. This technique was applied in seven consecutive patients. The defect was closed in all patients, however, only in three without complications. Indications, advantages and disadvantages of the treatment described are discussed. PMID- 9327282 TI - Closure of a post-traumatic maxillary defect. A case report. AB - A case of successful closure of a large maxillary defect is presented, using a free bone graft, rigid fixation and hydroxylapatite particles. PMID- 9327284 TI - Magnetic resonance imaging of the temporomandibular joint after functional treatment of bilateral condylar fractures in adults. AB - The position and functioning of discs in ten adult patients, whose bilateral condylar fractures were treated following a nonsurgical protocol, were investigated by means of magnetic resonance imaging. In seven temporomandibular joints (TMJs), where the condylar fragments were situated in the confines of the glenoid fossa, the discs were not displaced and were functioning normally. In 13 TMJs, where the condylar fragments were medially dislocated out of the fossa, the discs were displaced together with the condylar head but moved forward together with the condyle during mouth opening. This relationship between disc and condyle appears to play an important role in the reestablishment of function using nonsurgical treatment in patients with condylar fractures. PMID- 9327283 TI - Mandibular fractures following third molar extraction. A retrospective clinical and radiological study. AB - Twelve patients with 13 mandibular fractures following third molar extraction were treated in our Department between 1980 and 1995. Clinical and radiographic data relating to these patients were analysed retrospectively to determine complication characteristics. Only one fracture occurred during surgery. The first week after the operation was found to be most critical for fracture occurrence. A "cracking" sound from the jaw while eating frequently indicated that a fracture had occurred, but repeated radiographic examination may be necessary for definitive diagnosis. Most teeth removed belonged to Groups II/III according to PELL & GREGORY's classification, indicating partial or total impaction of the tooth and a narrow space in the retromolar triangle. In all cases, tooth roots were superimposed on or adjacent to the inferior alveolar canal. Clinically, the age of the patient seemed to be a common predisposing factor, and patients older than 30 to 40 years were considered to be a risk group. Regardless of the degree of impaction and tooth position, the fracture type seen was uniform, i.e. preangular location and a typical course of the fracture line. After adequate fracture treatment, no further complications were noted. PMID- 9327285 TI - Modified osteosynthesis for condylar neck fractures in atrophic mandibles. AB - A fixation system combining both plate and lag screw osteosynthesis for condylar neck fractures of the mandible is described. The currently available device is adapted in that the lag screw is inserted in the lateral cortical bone of the condylar segment instead of the cancellous bone alone. PMID- 9327286 TI - The combined use of endosteal implants and iliac crest onlay grafts in the severely atrophic mandible: a longitudinal study. AB - Thirteen patients, who received an onlay bone graft augmentation to their severely atrophic mandible in combination with a simultaneous implant insertion, were studied prospectively. A reproducible method that allowed for accurate assessment of graft resorption, consisting of lateral and oblique-lateral cephalometric radiographs in combination with an image analysis system, was used to assess the resorption rate in all patients and between subgroups of patients, according to selected patient and treatment characteristics. Thirty implants were placed, none of which were lost, and all patients expressed satisfaction after a mean observation period of 877 days. A mean resorption rate of 36% of the grafted bone height occurred, mainly during the first year and with some degree of individual variance. After three years the resorption had virtually stopped. No statistically significant differences between any of the subgroups of patients could be distinguished. Peri-implantitis was found around eight implants in seven patients. It is concluded that the described surgical technique should be used only if there are stringent indications. PMID- 9327287 TI - Interposed bone grafts to accommodate endosteal implants for retaining mandibular overdentures. A 1-7 year follow-up study. AB - The results are reported on 34 edentulous patients, who underwent interposed autogenous bone graft augmentation in the symphysis of the mandible, combined with bone-hydroxylapatite onlay augmentation of the area posterior to the mental foramina. Two to four implants were placed in the grafted symphysis after 3-5 months. An overdenture was constructed three months later. The follow-up period ranged from one to seven years. An average loss of mandibular bone height of 10 13% was observed. The data provided no evidence of a further time-dependent resorption from two-and-half to seven years. PMID- 9327288 TI - Clinical manifestations and diagnostic approach to metastatic cancer of the mandible. AB - In a 12-month period, metastatic cancer was diagnosed in eight patients. Six of them presented with pain mimicking toothache, temporomandibular joint disorders or trigeminal neuralgia, while two showed osteopenic bone lesions in the panoramic radiography, and perimandibular swelling. Anesthesia of the lower lip was the only common clinical feature. In seven of the eight patients, a whole body bone scintigraphy and single photon emission computed tomography (SPECT) of the skull in combination with a whole body and SPECT anti-granulocyte (Tc-99m MAK 250/183) bone marrow scintigraphy was performed. One patient did not have combined scintigraphy performed secondary to severe systemic illness. In six of the seven, the results were conclusive for a metastatic bone lesion. Biopsies confirmed three patients to have a previously unrecognized primary cancer, one patient to have previously unrecognized recurrent cancer, and three patients to exhibit new metastatic spread of an already diagnosed cancer. Histology revealed breast, lung, renal cancer and a malignancy of inconclusive origin. In the remaining patient, combined scintigraphy suggested osteomyelitis, yet biopsy revealed a prostate cancer metastasis with acute inflammatory cell infiltration. Thus, the scintigraphy pattern of a hot spot in the bone scan and a cold lesion in the bone marrow scintigraphy is highly suggestive of a mandibular metastasis, if accompanied by anesthesia of the lower lip. PMID- 9327289 TI - A method for the bony and dental reconstruction of the maxilla in dentate patients. AB - Reconstruction was carried out on eleven patients using a vascularized full thickness calvarial bone flap following partial maxillectomy. The donor site was covered with a split calvarial bone graft. Intraorally a mucosal transposition flap was used to cover the graft. Six months later implants were inserted and were allowed to heal for three months before dental rehabilitation began. No serious complications were encountered. PMID- 9327290 TI - Rhabdomyosarcoma masked by orbital trauma. AB - An orbital embryonal rhabdomyosarcoma is reported. The diagnosis was delayed by several weeks due to a concurrent history of repeated trauma. PMID- 9327291 TI - Ki-67 antigen in ameloblastomas: correlation with clinical and histological parameters in 54 cases from Kenya. AB - The aim of this study was to assess the cell proliferation in ameloblastomas and to correlate this with clinical features and histology. Immunohistochemistry with Ki-67 monoclonal antibody was performed on fresh tissue from 54 ameloblastomas. A labelling index (LI) was calculated by expressing the percentage of Ki-67 positive cells. There was no significant correlation between LI and clinical features: age, sex or tumour size. Follicular ameloblastomas had significantly higher LI (5.0 +/- 0.5; mean +/- SEM) than plexiform tumours (3.2 +/- 0.6; P < 0.05). Plexiform ameloblastomas from the anterior mandible had a significantly lower LI (1.8 +/- 0.5) than those from the posterior (3.9 +/- 0.8; P < 0.05). LI was higher in squamous arcades (6.4 +/- 3.1%) than in epithelial cords and cysts (1.4 +/- 1.3%; P < 0.001). These results suggest that LI correlates most closely with the histological pattern of the epithelium of ameloblastoma, both within and between different tumours. PMID- 9327292 TI - Stereophotogrammetric assessment of the effect of tenoxicam on facial swelling subsequent to third molar surgery. AB - The present study was undertaken in order to evaluate the effects of tenoxicam (Tilcotil), 20 mg daily, versus placebo on the inflammatory response induced in patients undergoing surgical removal of bilateral lower third molar teeth, using stereophotogrammetry to assess the degree of swelling. The results indicated that tenoxicam (20 mg/day) was no more effective than a nonactive placebo in modifying swelling in the first two postoperative days. PMID- 9327293 TI - A three-dimensional study of bending and torsion moments for different fracture sites in the mandible: an in vitro study. AB - The aim of the study was to determine and compare bending and torsion moments across mandibular fractures, for different positions of the bite point and different sites of the fracture. Three identical resin mandibles, each with a single fracture, were used. The fracture sites were in the angle, body and symphyseal regions. A polyethylene bone plate was used for fixation. Simulated bite forces were applied at 13 bite points. For each bite point, the displacements of the fragments were registered and converted into bending and torsion moments across the fracture. Positive bending moments were defined as those moments that caused compression at the lower border and tension at the alveolar side of the mandible; negative bending moments did the opposite. Angle fractures had relatively high positive bending moments. Body fractures had positive as well as negative bending moments and the highest torsion moments. Symphyseal fractures had negative bending moments only and relatively high torsion moments. It was found that angle, body and symphyseal fractures each have a characteristic load pattern. These load patterns should play a decisive role in the treatment of mandibular fractures with regard to number and positioning of plates. PMID- 9327294 TI - Low dosage of native allogeneic bone morphogenetic protein in repair of sheep calvarial defects. AB - A sheep skull trephine defect model was used to test the efficacy of allogeneic partially purified sheep bone morphogenetic protein (sBMP), extracted using a low cost alternative technique based on 60% ammonium sulphate saturation of the guanethidine-HCI extract of pulverized bone matrix. Eight mg of partially purified sBMP was implanted in six 22-mm right-side sheep calvarial critical-size defects trephined in the diploe area using a midline incision; left-side defects implanted with an equal amount of type IV collagen served as controls. After 16 weeks the sheep were killed and the defects removed. Formation of new bone was evaluated using radiomorphometry and histomorphometry. The healing percentage in sBMP-implanted defects was 60.8 +/- 8.1% and in controls 49.8 +/- 6.7% (P < 0.05) as assessed by radiomorphometry. In cross-sectional histomorphometry, newly formed bone regenerated 50.9 +/- 15.1% in the defects with sBMP and 16.1 +/- 10.6% in controls (P < 0.01). The good result, considering the low dosage of sBMP, can be explained by the strong osteoinductivity and low immunogenicity of native allogeneic sBMP. PMID- 9327295 TI - Has the TMI prosthetic protocol been changed? PMID- 9327296 TI - Operant schedule transformations and human behavioral momentum. AB - Behavioral momentum, which can be defined as the persistence of behavior under altered environmental contingencies, is derived from Newtonian physics and operant psychology, and has relevance to behavior therapy in terms of shaping strong behaviors and ensuring effective relapse prevention strategies. The present study investigated whether changing operant schedule contingencies affects how humans respond to different stimuli when reinforcement density is systematically manipulated. Fifteen subjects participated in a computer study, in which each of two keys in a baseline condition was associated with the same schedule of reinforcement, multiple variable interval schedules, the only difference being that one reinforcer was ten times larger than the other. After six sessions, the contingency schedule changed to either an extinction condition, a variable time schedule, or a changed variable interval schedule to assess how 'subjects' responses persisted when reinforcement contingencies were systematically changed. Results of this study were found to be consistent with the general predictions of behavioral momentum. Subjects not only biased responding in favor of the more densely reinforcing key, but when contingencies changed, subjects showed continued biased responding. Implications for behavioral momentum for behavior modification and behavior therapy are discussed, and it is concluded that behavioral momentum has significant implications for designing new and comprehensive behavior change programs. PMID- 9327297 TI - The transfer of avoidance evoking functions through stimulus equivalence classes. AB - Recent research in the area of stimulus equivalence suggests that transfer of function via members of stimulus equivalence classes may have relevance to human emotional responding and the development and generalization of certain psychological disorders. This study investigated the transfer of avoidance evoking functions through equivalence classes. Eight subjects were trained in the necessary relations for two-four member stimulus equivalence classes to emerge. Next, using an on-baseline classical conditioning procedure, one member of one class was paired with shock while one member of the other class was presented without shock. Then, while subjects engaged a key-press task, a differential, signalled avoidance task was introduced wherein shock was avoided if a response occurred to the stimulus previously associated with shock. The remaining stimuli from both classes were then presented. The behavior of all eight subjects showed the differential transfer of the avoidance evoking function. The clinical and theoretical implications of the results are discussed. PMID- 9327298 TI - Collegial support and barriers to behavioral programs for severe mental illness. AB - Previous investigations have identified staff beliefs about barriers to implementing behavioral interventions in programs for persons with severe mental illness. One of these barriers, institutional constraints, was found to be associated with collegial support; i.e., staff who report more collegial support were less likely to endorse institutional constraints. The purpose of this study was to determine how the components of collegial support were associated with beliefs about institutional constraints. Fifty-six staff members completed measures of staff opinions about barriers to implementing behavior therapy, satisfaction with collegial support, source of support, and functions of support. Results suggested that collegial support is significantly associated with co worker and supervisor support, but not the support of family and friends. Endorsing institutional constraints was inversely associated with the support of co-workers and supervisors; institutional constraints were positively associated with the support of family and friends. Endorsing institutional constraints was also inversely associated with the sense that others rely upon the individual for their well-being. Implications of these findings for diminishing barriers to behavioral interventions are discussed. PMID- 9327299 TI - Cognitive-behavioral remediation of problem solving deficits in children with acquired brain injury. AB - The inability to problem solve can have a deleterious impact on a student's academic performance and social adjustment. Children with an acquired brain injury (ABI) are at risk for deficits in problem solving skills. This case study and series of multiple baseline experiments examined the effects of a multi component cognitive-behavioral training program on the remediation of problem solving deficits in five children with ABI. Results indicated that the training program resulted in a substantial decrease in errors on a computerized problem solving task used to monitor problem solving performance during baseline and treatment. In addition, significant improvements were found on two of four standardized measures of problem solving abilities. Finally, students, parents and teaching staff reported a high degree of satisfaction with and generalization of the training program. PMID- 9327300 TI - An instrument for measuring staff's knowledge of behavior management principles (KBMQ) as applied to geropsychiatric clients in long-term care settings. AB - Direct-care staff in long-term care institutions for the elderly are responsible for implementing behavioral treatment programs for an increasing number of patients with behavioral and psychiatric problems. Because the quality of life for these patients is a function of the quality of care provided, maximizing staff's behavioral competencies could potentially improve patients' quality of life. Behavioral competence requires an understanding of behavioral principles. To assess direct-care staff's knowledge of behavioral principles as applied to geropsychiatric settings the KBMQ, a 30-item multiple choice instrument, was designed. The psychometric properties were tested with 203 staff. Results suggest that the instrument is reliable and valid. Potential uses and limitations are discussed. PMID- 9327301 TI - Comparison of specific and nonspecific factors in a group cognitive therapy for depression. AB - Research into the efficacy of psychotherapy has often reported equivalence in treatment outcome when comparing different therapies. These findings have been interpreted as evidence for what are variously termed placebo, common or nonspecific processes. We suggested that this issue is best examined in comparison of specific and nonspecific processes in the action of a specified therapy and disorder. No comparisons of this nature have yet been reported in relation to cognitive therapy for depression. This study compared specific processes (automatic thoughts and dysfunctional attitudes) and major common processes (satisfaction with therapy and client evaluation of therapist) in the action of a group cognitive therapy for depression. Sixty patients suffering from major depression received a 12 week course of group cognitive therapy. Results from hierarchical regression suggested that the specific processes of cognitive therapy were more associated with reduction in depression than common processes which contributed to the prediction of reduction in depression via specific processes. PMID- 9327302 TI - How specific are specific phobias? AB - To study the generality of fears among specific phobic individuals and controls, 31 individuals with a DSM-IV diagnosis of specific phobia (natural environmental type: n = 13; blood-injection-injury type: n = 10; and situational type: n = 8) and 33 never mentally ill control subjects participated in an interview and questionnaire study. Based on subjects' fear ratings on the Fear Survey Schedule, subjects were classified as either positive or negative with regard to fear categories that correspond to the five diagnostic subtypes of specific phobia. Phobics showed overall a more generalized form of fear than controls. Furthermore, situational fears were more common among specific phobics who did not meet criteria for specific phobia, situational type, than among controls. These results add to the literature on the functional relationship among different fears and suggest that specific phobias are not as "specific" as is implied by the current diagnostic system. PMID- 9327303 TI - A simplified method of toilet training adults in residential settings. AB - A simplified version of Azrin and Foxx's method of toilet training was evaluated in an adult with profound mental retardation. An ABAB reversal design was used to evaluate intervention effects. Results indicated that the procedure was effective in reducing toileting accidents and in increasing appropriate urinations. Additionally, continence was maintained at a 3 month follow-up evaluation. PMID- 9327304 TI - Welch's comments on Shapiro's walk in the woods and the origin of eye movement desensitization and reprocessing. AB - Welch's (Journal of Behavior Therapy and Experimental Psychiatry, 27, 175-179, 1996) response to Rosen's (Journal of Behavior Therapy and Experimental Psychiatry, 26, 121-122, 1995) limited study on the origin of eye movement desensitization and reprocessing (EMDR) does not resolve how best to interpret what Shapiro experienced during her reported walk in the woods. References cited by Welch actually argue against the conclusions he advances. PMID- 9327305 TI - Feeling the heat. PMID- 9327306 TI - Prolonged rewarming after hypothermic cardiopulmonary bypass does not attenuate reduction of jugular bulb oxygen saturation. AB - OBJECTIVE: This study investigates the effects of rapid versus graded rewarming on decreases in jugular bulb oxygen saturation (SjO2) during cardiopulmonary bypass (CPB) in a prospective nonrandomized and nonblinded design. SETTING AND PARTICIPANTS: At the Department of Anesthesiology (University Hospital Eppendorf, Germany), 28 patients (ASA III) undergoing coronary artery bypass graft were investigated. INTERVENTION: CPB was managed according to alpha-stat conditions during moderate hypothermia (27 degrees C). In group 1 (n = 17), rewarming was performed by increasing the perfusate temperature to 36 degrees C within 7 minutes, in group 2 (n = 11) within 15 minutes. MEASUREMENTS AND MAIN RESULTS: SjO2 was measured by a fiberoptic catheter placed in the right jugular bulb. Data were recorded before and 40 minutes after the start of rewarming every 5 minutes. During rewarming of CPB, SjO2 was decreased to 43 +/- 7% in group 1 and to 44 +/- 4% in group 2. In groups 1 and 2, the maximum reduction of SjO2 occurred 17 minutes and 30 minutes after start of rewarming, respectively. The delayed reduction of SjO2 in group 2 correlated strongly with the prolonged increase in jugular bulb temperature. CONCLUSION: The current data show that slow rewarming does not attenuate reductions of SjO2. This suggests that the reduction of SjO2 during rewarming of CPB is not a function of the rewarming speed but is strongly correlated with the increase in jugular bulb temperature, with a maximum effect just before reaching normothermia of the brain. PMID- 9327307 TI - Neurologic findings in coronary artery bypass patients: perioperative or preexisting? AB - OBJECTIVES/DESIGN: This prospective study compares the incidence of preexisting neurologic findings in elective cardiac surgery patients presenting with and without coronary atherosclerosis. SETTING: This single-center study was conducted at a tertiary care hospital. PARTICIPANTS/INTERVENTIONS: After Review Board approval and obtaining written informed consent, 11 patients undergoing valvular heart surgery, 9 patients undergoing similar valvular procedures with concomitant coronary artery bypass surgery, and 4 patients undergoing coronary artery bypass surgery alone were enrolled. Preoperatively, all patients underwent a structured neurologic assessment, and the latter four additionally had preoperative magnetic resonance imaging. MEASUREMENTS AND MAIN RESULTS: The patients, 9 of 24 of whom were female, were aged 46 to 78 years and, other than ischemic heart disease, had medical histories that were similar between groups, with the exception of one patient having scleroderma. None of the patients had a clinical history of neurologic or cerebrovascular disease. Nine percent (1 of 11) of the valve-only patients showed subtle preoperative neurologic abnormalities, compared with 89% (eight of nine) of the valve patients having concomitant coronary surgery and 100% (four of four) of coronary artery bypass-only patients. Additionally, brain imaging scans of all four coronary bypass patients showed nonspecific changes reported as scattered punctate areas of high signal less than 3 to 4 mm in diameter. CONCLUSION: This survey shows that both subtle neurological abnormalities and magnetic resonance imaging lesions can be found in a high percentage of patients with coronary atherosclerosis. Furthermore, this study indicates that without a standardized preoperative neurological examination, postoperative neurologic dysfunction cannot necessarily be ascribed to perioperative events. PMID- 9327308 TI - The appearance of S-100 protein in serum during and immediately after cardiopulmonary bypass surgery: a possible marker for cerebral injury. AB - OBJECTIVE: To investigate the appearance and elimination of brain-specific S-100 protein in serum during and immediately after cardiopulmonary bypass. DESIGN: Prospective study. PARTICIPANTS: Twenty-nine patients undergoing elective cardiac surgery. INTERVENTIONS: Twenty-seven patients were operated on for coronary artery disease; two patients had valve replacement. Serial measurements of S-100 in arterial blood during and up to 48 hours after cardiopulmonary bypass were made. MEASUREMENTS AND MAIN RESULTS: The perioperative and postoperative course was uneventful in 25 patients, with no clinical signs of neurologic complications. S-100 was not detected before extracorporeal circulation was started. Detectable concentrations (detection limit, 0.2 microgram/L) appeared in serum after 10 minutes of perfusion and reached maximum levels, 2.43 +/- 0.3 micrograms/L, at the end of bypass. The levels then declined with elimination t1/2 of 2.2 hours. Only two patients had detectable concentrations of S-100 48 hours after the end of bypass. In four patients who developed clinical signs of cerebral injury, levels of S-100 were significantly higher at the end of bypass and 24 hours after the end of bypass. CONCLUSIONS: Cardiopulmonary bypass initiates a release of brain-specific S-100 to the systemic circulation. The release and elimination of S-100 seem to follow a reproducible pattern in patients with no signs of cerebral injury. In patients who developed cerebral injury, the concentrations of S-100 in blood were increased, thus suggesting that S-100 may be a usable marker for cerebral injury after extracorporeal circulation. PMID- 9327309 TI - The intraoperative assessment of ascending aortic atheroma: epiaortic imaging is superior to both transesophageal echocardiography and direct palpation. AB - OBJECTIVES: To determine the optimal method for detecting ascending aortic atheroma intraoperatively by comparing manual palpation by the operating surgeon, intraoperative transesophageal echocardiography, and epiaortic ultrasound (linear and phased-array imaging); and to assess risk factors for severe aortic atheroma. DESIGN: A longitudinal prospective study. Assessment of the atheroma by manual palpation was blinded to the results of the ultrasound images. SETTING: The study was performed in a single university tertiary referral hospital. PARTICIPANTS: One hundred consecutive patients undergoing coronary bypass or valve surgery were studied after their written, informed consent. INTERVENTIONS: Potential risk factors were evaluated by both a patient questionnaire and examination of prior hospital records. The ascending aorta was assessed by the following methods: manual palpation by the operating surgeon, intraoperative transesophageal echocardiography, and epiaortic ultrasound (linear and phased-array imaging) performed by an echocardiologist. For analysis, the ascending aorta was divided into three equal segments: proximal, mid, and distal, corresponding to regions of different operative manipulations. MEASUREMENTS AND MAIN RESULTS: Age older than 70 years and hypertension were significant risk factors for severe ascending aortic atheroma with adjusted odds ratios of 3.3 (95% CI, 1.2 to 9.3) and 3.9 (95% CI, 1.3 to 12.0), respectively. There was no significant difference in atheroma detection between the two ultrasonic epiaortic probes in any segment; however, epiaortic probes were superior to manual palpation in all segments and also superior to transesophageal echocardiography in the mid and distal segments of the ascending aorta. CONCLUSIONS: Age older than 70 years and hypertension are significant risk factors for severe ascending aortic atheroma. Intraoperative detection of ascending aortic atheroma is best achieved by epiaortic ultrasound with either a linear or phased array transducer. Transesophageal echocardiography is an insensitive technique for evaluation of mid and distal ascending aortic atheroma and, therefore, of little value in guiding surgical manipulations such as cross-clamping. PMID- 9327310 TI - Forced-air warming is no more effective than conventional methods for raising postoperative core temperature after cardiac surgery. AB - OBJECTIVE: To determine whether postoperative forced-air warming of cardiac bypass patients in the intensive care unit (ICU) results in faster rate of warming and improved outcomes compared with more conventional ICU warming methods. DESIGN: Prospective randomized effectiveness study. SETTING: Three hundred fifty-bed university-affiliated hospital. PARTICIPANTS: Sixty consenting randomized patients from a consecutive series of 84 patients undergoing routine adult cardiac surgery. INTERVENTIONS: One group of patients received usual patient care, which includes warm blankets and overhead heat lamps. Patients in the other group were placed under forced-air warming devices on arrival in the ICU. Sixty consenting patients (30 in each group) were randomly assigned to one or the other method of warming. The remaining 24 patients refused randomization and self-selected a treatment group. MEASUREMENTS AND MAIN RESULTS: Results are presented for the randomized groups. Core temperature, measured by pulmonary artery catheter thermistor, increased in both groups at the rate of 0.25 degree C per hour. No statistically or clinically significant differences were found between the group for whom the warming device was used and the standard care group in the incidence of postoperative cardiac arrhythmia, duration of time in the ICU, or any other clinical variable. CONCLUSIONS: There is no evidence from this study to warrant use of forced-air warming devices for the care of postoperative cardiac surgical patients in the ICU. PMID- 9327311 TI - Comparison of the effects of sodium nitroprusside and isoflurane during rewarming on cardiopulmonary bypass. AB - OBJECTIVES: Afterdrop in core temperatures after discontinuation of cardiopulmonary bypass (CPB) is reported to be a sign of inadequate total body rewarming on CPB. The purpose of this study was to compare the effects of three different drug regimens on hemodynamic stability and the uniformity of rewarming during the rewarming period of CPB. DESIGN: This prospective randomized study was performed in the Anesthesiology Department of the University of Istanbul. PARTICIPANTS: Sixty-six patients undergoing uncomplicated valve replacement and aortocoronary bypass grafting surgery were studied. INTERVENTIONS: Anesthesia was maintained with isoflurane and fentanyl infusion during the prebypass and the postbypass periods. Patients were allocated into three groups by the initiation of CPB. Group 1 (n = 22): fentanyl infusion + diazepam + sodium nitroprusside (SNP) in the rewarming period), group 2 (n = 22): fentanyl infusion + isoflurane, group 3, control (n = 22): fentanyl infusion + diazepam. Rectal, esophageal, and forearm temperatures were monitored throughout the study. MEASUREMENTS AND MAIN RESULTS: None of the durational and temperature data showed significant differences between groups 1 and 2. In the control group, afterdrop in esophageal temperature was significantly higher than groups 1 and 2 (group 1: -1.4 +/- 0.9 degrees C, group 2: -1.44 +/- 0.8 degrees C, group 3: -2.1 +/- 0.65 degrees C). In group 1, the number of patients whose mean arterial pressure (MAP) decreased below 45 mmHg was significantly higher than group 2 (p = 0.002). Mean SNP infusion rate and mean isoflurane concentration during the rewarming period were calculated as 1.55 +/- 0.8 micrograms/kg/min and 0.775 +/- 0.27%, respectively. CONCLUSIONS: Isoflurane produced more stable hemodynamic conditions than SNP during the rewarming period, improved the uniformity of rewarming, and permitted earlier extubation in the intensive care unit (ICU). It is concluded that isoflurane alone is capable of fulfilling the anesthesia needs during hypothermia and the rewarming period of CPB. PMID- 9327312 TI - Proinflammatory cytokine release during pediatric cardiopulmonary bypass: influence of centrifugal and roller pumps. AB - OBJECTIVE: It has been proposed that nonocclusive centrifugal pumps may elicit less blood cell trauma and hence a reduced inflammatory response than standard roller pumps. However, there have been no reports describing the impact of such pumps on proinflammatory cytokine release in pediatric cohorts. DESIGN: A prospective randomized study was undertaken. SETTING: A regional cardiothoracic center of a university hospital. PARTICIPANTS: Thirty-four pediatric patients undergoing cardiopulmonary bypass (CPB) for the correction of complex congenital heart defects were recruited. INTERVENTIONS: Either standard twin roller (n = 17), or centrifugal vortex (Biopump, Medtronic Biomedicus Inc, MN) (n = 17) blood pumping. MEASUREMENTS AND MAIN RESULTS: Venous blood was drawn (1) on induction of anesthesia, (2) 5 minutes on bypass, (3) end of CPB, (4) 30 minutes post protamine, (5) 2 hours and (6) 24 hours postoperation. Neutrophil count, level of plasma leukocyte elastase, terminal complement complex (C5b-9); interleukin-6 (IL 6) and interleukin-8 (IL-8) were increased during and after CPB compared with the postinduction baseline. C5b-9 levels in both groups peaked at the end of CPB before returning to baseline at 24 hours: (median [range]), 564 (16 to 1,136) ng/mL in centrifugal group versus 508 (0 to 1,128) ng/mL in the roller group. IL 6 in both groups reached its peak level at 2 hours postprotamine (208 [98 to 411] pg/mL in centrifugal versus 205 [60-327] pg/mL in the roller group), before coming back to baseline at 24 hours. Plasma leukocyte elastase and IL-8 reached their maximum level 15 minutes after protamine administration: 215 (64 to 375) pg/mL in centrifugal versus 235 (87 to 410) pg/mL in roller group; and 700 (90 to 5,925) ng/mL versus 362 (120 to 3,400) ng/mL, respectively. CONCLUSIONS: The current study confirms the proinflammatory nature of pediatric CPB surgery, but failed to show a significant advantage of centrifugal pumping over roller perfusion in terms of the inflammatory response. PMID- 9327313 TI - Hemodynamic benefit of optimizing atrioventricular delay after cardiopulmonary bypass. AB - BACKGROUND: Shortening of atrioventricular delay (AVD) by sequential cardiac pacing has been proposed to improve hemodynamics in patients with end-stage heart failure. In addition, optimization of prolonged AVD may be associated with a decrease of presystolic mitral insufficiency. The aim of this study was to explore the incidence of prolonged AVD during the early postcardiopulmonary bypass (CPB) period and to evaluate the hemodynamic benefit of its shortening by using sequential cardiac pacing. METHODS: Fifty consecutive patients scheduled for coronary artery bypass grafting were prospectively screened. AVD was measured immediately after separation from CPB. Patients presenting with AVD greater than or equal to 200 ms entered the study. Sequential cardiac pacing was introduced with programmed AVD starting at 80 ms and randomly increased by steps of 20 ms until resumption of native anterograde atrioventricular node conduction. Cardiac index (CI) was derived from transesophageal echocardiographic data during each step of this procedure. RESULTS: Nineteen patients were included. Median native AVD was 220 ms. Median optimal AVD was 140 ms. Mean native CI (CI-nat) was 2.59 +/- 0.42 L/min/m2. Mean optimal CI (CI-opt) was 3.12 +/- 0.45 L/min/m2. CI-opt/CI nat was 1.20 +/- 0.07. CI-opt/CI-nat was significantly inversely correlated with preoperative left ventricular ejection fraction (r = -0.83). CONCLUSIONS: Prolonged AVD is a common occurrence after CPB. Its artificial shortening by sequential cardiac pacing is always associated with a significant increase of CI. The magnitude of this hemodynamic improvement is inversely correlated with preoperative left ventricular ejection fraction. PMID- 9327314 TI - A randomized multicenter double-blind comparison of urapidil and ketanserin in hypertensive patients after coronary artery surgery. AB - OBJECTIVES: To compare the hemodynamic responses, safety, and efficacy of urapidil and ketanserin in hypertensive patients after coronary artery surgery. DESIGN: Randomized double-blind study. SETTING: Multi-institutional. PARTICIPANTS: One hundred twenty-two patients undergoing elective coronary artery surgery. INTERVENTIONS: When hypertension (defined as mean arterial pressure > 85 mmHg) developed within the first 2 hours after arrival in the intensive care unit, patients received urapidil (n = 62) or ketanserin (n = 60) to reach a mean arterial pressure between 65 and 75 mmHg. Urapidil was administered by repeated bolus injections (25 to 125 mg) followed by a continuous infusion of maximally 50 micrograms/kg/min. Ketanserin was administered by repeated bolus injections (10 to 50 mg) followed by a continuous infusion of maximally 4.0 micrograms/kg/min. MEASUREMENTS AND MAIN RESULTS: A complete hemodynamic profile was determined at baseline and at 30 and 60 minutes after start of study medication. In the urapidil group, mean arterial pressure (+/-SD) decreased significantly from 100.6 +/- 12.4 mmHg at baseline to 74.6 +/- 12.1 mmHg at 30 minutes and 73.5 +/- 13.8 mmHg at 60 minutes. In the ketanserin group, mean arterial pressure decreased significantly from 98.7 +/- 10.7 mmHg at baseline to 83.5 +/- 16.8 mmHg at 30 minutes and 83.1 +/- 15.3 mmHg at 60 minutes. Between the groups, there was a significant difference in the degree of lowering mean arterial pressure at 30 and 60 minutes. Heart rate increased significantly by 5.8 +/- 12.7 (30 minutes) and 8.6 +/- 16.5 (60 minutes) beats/min in the ketanserin group. In the urapidil group, no changes in heart rate occurred. Cardiac output increased to the same extent (0.7 L/min) in both groups. Within and between the groups, there were no relevant changes in pulmonary filling pressures. The number of patients not responding adequately to the study medication (mean arterial pressure > 85 mmHg after 30 minutes despite the maximum doses of study medication) was comparable in both groups (9 [U] v 13 [K]). Adverse events attributable to the study medication occurred to a similar degree in both groups. In the patients treated with urapidil, a significantly higher incidence (32.3%) of hypotension (mean arterial pressure < or = 65 mmHg for more than 10 minutes) occurred after 60 minutes of continuous infusion. CONCLUSIONS: In contrast to ketanserin, urapidil did not increase heart rate. Urapidil was more effective in lowering arterial blood pressure than ketanserin. However, one third of the patients treated with urapidil developed hypotension after 60 minutes of continuous infusion. PMID- 9327315 TI - Oral transmucosal fentanyl citrate as an additional premedicant for adult cardiac surgery patients. AB - OBJECTIVE: To investigate the efficacy of a combination of midazolam and oral transmucosal fentanyl citrate (OTFC) as a preoperative medication for adult cardiac surgery patients compared with the use of midazolam alone. DESIGN: A randomized, prospective study. SETTING: University teaching hospital. PARTICIPANTS: Patients scheduled for elective coronary artery bypass surgery. INTERVENTIONS: All patients were given 50 micrograms/kg of midazolam intramuscularly in their rooms. Group I received 300 micrograms of OTFC Oralet (Anesta Corp, Salt Lake City, UT) if they weighed less than 70 kg and 400 micrograms of OTFC Oralet if they weighed more than 70 kg. Group II received a placebo Oralet. A radial artery catheter, two internal jugular venous catheters, and a pulmonary artery catheter inserted through one of the internal jugular catheters were placed in each study patient. Fentanyl was administered intravenously as a rescue drug. MEASUREMENTS AND MAIN RESULTS: Ninety percent of midazolam/OTFC patients reported feeling no pain during catheter placement, compared with 50% of midazolam/placebo patients. Fifty percent of the placebo group required fentanyl supplement of 50 micrograms intravenously because of complaints of pain, compared with 10% of the OTFC group. The midazolam/OTFC group scored approximately 20% better than the placebo group in the independent observer score of patient analgesia and the anesthesiologist rating for ease of invasive catheter placement. No myocardial ischemic events were noted in either group as determined by electrocardiogram. All patients found the Oralet mode of delivery very acceptable. CONCLUSIONS: The OTFC Oralet provides effective analgesia and sedation when combined with midazolam for invasive catheter placement in adult cardiac surgery patients. The OTFC Oralet with its gradual onset lessens the possibility of overmedicating with fentanyl, and it offers a very acceptable mode of delivery for a preemptive analgesic. PMID- 9327316 TI - Comparison of isoflurane and midazolam as hypnotic supplementation to moderately high-dose fentanyl during coronary artery bypass grafting: effects on systemic hemodynamics and early postoperative recovery profile. AB - OBJECTIVE: The aim of this study was to compare isoflurane and midazolam as hypnotic adjuncts to moderately high-dose fentanyl during coronary artery bypass grafting (CABG) with regard to perioperative hemodynamics and early postoperative recovery profile. DESIGN: Prospective open randomized clinical trial. SETTING: Single university-affiliated medical center. PARTICIPANTS: Thirty patients scheduled for elective primary CABG were randomly divided into two groups receiving either isoflurane or midazolam as adjuncts to 50 micrograms/kg of fentanyl. INTERVENTION: Anesthesia was induced with intravenous fentanyl, 10 micrograms/kg, and midazolam, 0.1 mg/kg, and maintained with either isoflurane, 0.6% or midazolam, 0.1 mg/kg/hour, in intravenous infusion. Before the sternotomy, all patients received 30 micrograms/kg of fentanyl. Midazolam and isoflurane were stopped at the start of cardiopulmonary bypass (CPB). At rewarming, all patients received fentanyl, 5 to 10 micrograms/kg, and either isoflurane, 0.6%, or midazolam, at a reduced rate of 0.05 mg/kg/h. Changes in systolic blood pressure of more than 20% from baseline were first treated with vasoactive drugs. Hypertension was corrected with ketanserin, 10 to 20 mg; hypotension with ephedrine, 5 mg. For a mean blood pressure of less than 50 mmHg during CPB phenylephrine, 0.25 to 0.5 mg, was administered. When hypotension persisted despite a vasopressor, the administration of midazolam or isoflurane was stopped. Postoperatively, the patients were mechanically ventilated overnight and sedated with intermittent doses of fentanyl, 0.15 mg, and midazolam, 5 mg. MEASUREMENTS AND MAIN RESULTS: Routine five-lead electrocardiogram (ECG) and invasive hemodynamic monitoring using a pulmonary artery catheter were performed. The mean dose of fentanyl was 3.9 mg in the midazolam group versus 3.6 mg in the isoflurane group. There were no significant perioperative differences between groups in cardiac output, filling, or pulmonary artery pressures. Systolic blood pressure from the initial incision to CPB was lower in the isoflurane group compared with the midazolam group. During this interval, ketanserin was required in nine patients from the midazolam group at a mean dose of 26 mg, compared with only one patient in the isoflurane group. During and after CPB, there was no difference in ketanserin and in vasopressive/inotropic agent requirements. Temporary cessation of midazolam was required in four patients, for a mean of 34 minutes; whereas isoflurane was stopped in 10 patients for 36 minutes, mostly in the post-CPB period. Time to awakening and to extubation in the midazolam group (217 minutes and 19.5 hours) and the isoflurane group (193 minutes and 18.2 hours) were comparable. Between intensive care unit (ICU) admission and extubation, the patients in the midazolam group received 0.89 mg of fentanyl and 36.5 mg of midazolam compared with 0.98 mg of fentanyl and 36.2 mg of midazolam in the isoflurane group. There was a tendency for a higher postoperative pulmonary shunt and more severely impaired oxygenation in the isoflurane group. CONCLUSION: Midazolam supplementation to fentanyl required more frequent antihypertensive escape during the pre-CPB period than isoflurane. However, more frequent cessation of isoflurane caused by hypotension was needed in the post-CPB period. No difference in awakening and ICU discharge was found. PMID- 9327317 TI - Perioperative metoprolol reduces the frequency of atrial fibrillation after thoracotomy for lung resection. AB - OBJECTIVE: The association of atrial fibrillation with thoracic surgical procedures is well known, but nevertheless its cause is not well defined. Increased sympathetic activity may play a role in the development of atrial fibrillation, and reduced beta-receptor activity may be advantageous. The objective was to evaluate the effect of oral beta-blockade on the frequency of atrial fibrillation and to evaluate some possible causative factors. DESIGN AND SETTING: The study was prospective, randomized, and double-blind, and was conducted at Aarhus University Hospital. PARTICIPANTS: Thirty patients without previous or present cardiovascular history undergoing elective thoracotomy for lung resection. INTERVENTIONS: The patients received either 100 mg of metoprolol or placebo orally before surgery and once daily postoperatively. Anesthesia consisted of a thoracic epidural block combined with general intravenous anesthesia. Epidural morphine was continued postoperatively. MEASUREMENTS AND MAIN RESULTS: Patients were monitored with electrocardiograms (ECGs), capillary pulse oximetry, invasive hemodynamic monitoring, central venous oxygen saturation, arterial blood gases, serum electrolytes, and fluid balances. Atrial fibrillation developed in 23.3% of the patients, 6.7% after metoprolol compared with 40% in the placebo group. Atrial fibrillation developed a mean of 2.9 days postoperatively. The predominant hemodynamic findings were perioperative lower oxygen consumption and postoperative lower cardiac index after metoprolol. Patients developing atrial fibrillation had much higher oxygen consumption and postoperative cardiac index than other patients. CONCLUSION: Perioperative oral beta-blockade can reduce the frequency of atrial fibrillation without serious side effects. Increased sympathetic activity is one of the predominant factors in the cause of this complication. PMID- 9327318 TI - Evaluation of the efficacy of routine preoperative electrocardiograms. AB - OBJECTIVE: To evaluate the efficacy of routine preoperative electrocardiograms (ECG) in predicting perioperative cardiovascular complications in an essentially healthy population. DESIGN: Retrospective chart review. SETTING: The adult hospital of a large academic medical center. PARTICIPANTS: One thousand ASA class I and II adult patients undergoing a number of different elective surgical procedures. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The mean age of the population was 48 years (range, 18 to 88), and 53% were female. Fifty-seven percent of the population had a preoperative ECG, of which 56.5% were considered normal, 37.8% were abnormal, and 6.6% were considered borderline. Twenty-seven percent of the population presented with cardiovascular risk factors, and of these, 93.2% had an ECG performed. Seventy-three percent of patients had no cardiovascular risk factors, and of these, 44.5% had a preoperative ECG. Patients who had cardiovascular risk factors had significantly more abnormal ECGs than those without (51% v 26.1%,); however, there was no difference in the prevalence of perioperative events between the two groups. The positive predictive value of an abnormal ECG for a perioperative event was slightly greater for patients with cardiovascular risk factors than for those without (42.7% v 34.7%, respectively); however, this difference was not significant. In addition, a normal ECG was just as predictive as an abnormal one. CONCLUSIONS: Results of this study suggest that the practice of routine ECG screening for patients with no cardiovascular risk factors is a poor predictor of perioperative complications in this patient population. A review of the current criteria for ordering preoperative ECGs may reduce the number of unnecessary tests and improve cost-effectiveness. PMID- 9327319 TI - Coronary thrombosis associated with antithrombin-III deficiency. PMID- 9327320 TI - Indicators of fibrinolysis during cardiopulmonary bypass after exogenous antithrombin-III administration for acquired antithrombin III deficiency. PMID- 9327321 TI - Recognition and management of patients with antiphospholipid antibody syndrome undergoing cardiac surgery. PMID- 9327322 TI - Unexplained intraoperative hypotension, acute ischemic hepatitis, and pancreatitis associated with aortorenal bypass surgery. PMID- 9327323 TI - New concepts and therapies of adult respiratory distress syndrome. PMID- 9327324 TI - Case 4--1997. The role of ischemic preconditioning during minimally invasive coronary artery bypass surgery. PMID- 9327325 TI - Pro: prevention of neurologic dysfunction associated with cardiac surgery requires pharmacologic brain protection. PMID- 9327326 TI - Con: preventing stroke after cardiopulmonary bypass does not require pharmacologic neuroprotection. PMID- 9327327 TI - Failure of a Swan-Ganz catheter to show a pulmonary artery wedge trace. PMID- 9327328 TI - Benefits of neuraxial anesthesia in patients undergoing cardiac surgery. PMID- 9327329 TI - Improvement of cardiac function by allopurinol. PMID- 9327330 TI - Choice of double-lumen tube in Kartagener's syndrome. PMID- 9327331 TI - Anesthesia for DiGeorge's syndrome. PMID- 9327332 TI - Measurement of myocardial contractility. PMID- 9327333 TI - Regulation of mucosal immune responses: distinct antigens and antigen presenting cells. PMID- 9327335 TI - Interleukin-17 and its receptor. PMID- 9327334 TI - Host-pathogen interactions in the immunopathogenesis of Lyme disease. AB - The immunopathogenesis of Lyme disease is complicated and requires a thorough understanding of the interaction among the causative organism, Borrelia burgdorferi, its tick vector, and its mammalian hosts. In vitro, animal and human studies have shown that the organism is capable of adapting to and utilizing elements from its environment to establish infection and persist despite a inducing a strong immune response. Indeed, the immune response may be responsible for many of the symptoms associated with Lyme disease. It appears that humoral immunity plays the greatest role in clearance of the organism. Cytokines released by Th 1 or Th 2 subsets of CD4+ cells have been shown to play an important role in determining outcome of the disease in animal models possibly through their effects on immunoglobulin class switching. In the small percentage of patients who have treatment resistant chronic Lyme disease, autoimmune mechanisms may play a role in persistent disease. PMID- 9327336 TI - Rectal immunization for induction of specific antibody in the genital tract of women. AB - The purpose of the current study was to examine potential routes of vaccine administration for the induction of antigen-specific responses in the genital tract of women. Sixteen women were enrolled in this study, and the level of influenza-specific antibodies induced in the genital tract was measured after rectal or intramuscular immunizations. Both methods of administration induced significant increases in the concentration of flu-specific IgA found in cervical secretions within 28 days after vaccination. Initially flu-specific IgG antibodies were not induced in the genital tract by either route. As expected both IgA and IgG flu-specific antibodies were dramatically increased in serum after intramuscular vaccination. In contrast, rectal administration did not induce significant IgA responses, and only small flu-specific IgG increases in serum. Six months after administration, IgA flu-specific antibody concentrations were significantly higher than baseline levels in vaginal secretions and saliva isolated from both subject groups and flu-specific IgG concentrations in cervical secretions were high in the rectal immunization group. The long-term presence of both IgG and IgA antibody in genital secretions suggests that rectal immunization may be an effective method for induction of immune protection in the genital tract of women. PMID- 9327337 TI - Elevated serum cyclophilin levels in patients with severe sepsis. AB - Several cytokines are considered to be important mediators in the pathophysiology of sepsis. Cyclophilins (Cyps), the main binding proteins for the immunosuppressive drug cyclosporine A, have been suggested to function as cytokines. This study was conducted to determine (i) if serum Cyp levels were elevated in critically ill patients suffering from either sepsis or other life threatening diseases and (ii) if so, whether there was an association between Cyp levels and a certain diagnosis and/or outcome. Serum samples of 45 patients (22 severe sepsis, 23 other diagnoses) and 17 healthy controls were prospectively analyzed by an enzymatic assay using the ability of cyclophilins to catalyze cis/trans isomerisation of peptidylprolyl-peptide bonds (PPIase activity). In addition, western blotting was applied to differentiate both isoforms. PPIase activity was significantly higher in patients with severe sepsis than in patients with other diagnoses (P = 0.004) or in healthy subjects (P = 0.001). There was no difference between healthy subjects and other critically ill patients (P = 0.067). Elevated PPIase activity was associated with high mortality (P = 0.03). It is concluded that Cyps might play a role, probably as mediators in the pathophysiology of sepsis or as symptoms of diagnostic value. PMID- 9327338 TI - Altered IL-1 expression and compartmentalization in monocytes from patients with AIDS stimulated with Mycobacterium avium complex. AB - The pathophysiologic basis for the exuberant intracellular growth of Mycobacterium avium complex (MAC) in AIDS patients is unclear but may relate to altered expression of modulatory cytokines. Interleukin (IL)-1, IL-6, and TNF alpha expression by monocytes from AIDS patients and healthy subjects (HS) stimulated with isogeneic MAC strains (SmT, smooth-transparent, virulent; SmD, smooth-domed, avirulent) was examined. Spontaneous cytokine production was not observed in patients with AIDS. MAC strains induced less IL-1 alpha and IL-1 beta release in AIDS patients than HS (P < 0.05). The ratio of cell-associated to supernatant IL-1 alpha also was increased in AIDS patients (P = 0.03). IL-1 beta mRNA expression paralleled protein release in either group of subjects. In both HS and AIDS patients, stimulation with SmD induced more IL-1 and TNF-alpha release by monocytes compared to SmT. In AIDS patients, SmD also induced greater IL-6 release than SmT (P < 0.01). Alterations in monocyte expression and compartmentalization of the regulatory cytokines IL-1 and IL-6 may enhance bacterial replication and contribute to the pathogenesis of MAC infection in AIDS. PMID- 9327340 TI - Molecular characterization of complement components (C3, C4, and factor B) in human saliva. AB - A molecular analysis of complement components (C3, C4, and factor B) in human saliva was performed by SDS-PAGE and immunoblotting. Complement C3 was detected as a molecule composed of a 115-kDa alpha-chain linked to a 70-kDa beta chain by disulfide bonds, and C3 levels ranged from 0.52 to 15.0 micrograms/ml (n = 15). C4 was detected as a triple-chain molecule (98-kDa alpha chain, 73-kDa beta chain, and 33-kDa gamma chain) linked by disulfide bonds, and C4 levels ranged from 0.086 to 4.8 micrograms/ml. Factor B was detected as a 100-kDa single chain, and factor B levels ranged from 0.042 to 0.62/microgram/ml. The sizes and subunit structures of the complement components in human saliva were compatible with those reported in human serum. The results of a hemolytic assay indicated that the complement molecules in human saliva were functionally active. These complement components may participate in the local immune and inflammatory responses in the oral cavity. PMID- 9327339 TI - Phenotypic characterization of cytokine expression in patients with IgA nephropathy. AB - To identify the cytokines that play a relevant role in the pathogenesis of IgA nephropathy, we analyzed and compared the gene expression of proinflammatory cytokines, immuno-regulatory cytokines, and growth factors in peripheral blood mononuclear cells (PBMC). Expression of IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma, TGF-beta, TNF-alpha, and PDGF was examined in 28 patients with IgA nephropathy (IgAN), 20 patients with non-IgA mesangial proliferative glomerulonephritis (mesPGN), and 19 healthy controls. Compared with healthy controls, a significant number of IgAN and mesPGN patients showed increased expression of IL-1 beta, IL-4, IL-10, IL-12, and IFN-gamma. The cytokine profile of renal tissue of 10 IgAN and 5 mesPGN biopsies was simultaneously analyzed and compared with that of PBMC. The proinflammatory IL-1 alpha and growth factor PDGF-B were expressed more in renal tissues than in PBMC. Furthermore, the renal profile of IL-alpha, IFN-gamma, and TNF-alpha expression was associated with the expression of IFN-gamma in PBMC. The serum level of IFN gamma of IgAN correlated significantly (P = 0.0003) with that of IL-12, suggesting a potential role for cross-stimulation. More importantly, expression of IFN-gamma in PBMC and the elevated serum level correlated with the decline in glomerular filtration rate (P = 0.0012) and severity of renal histopathologic grade. To elucidate the role of leukocytes in renal cytokine expression, surface markers of T cells (CD3), monocytes (CD14), natural killer cells (CD16), and B cells (CD19) were also examined in renal tissues. The prominent renal expression of CD3, CD14, and CD16 implicates the leukocytes as the major source of proinflammatory cytokines in IgAN. Collectively, these findings indicate that IFN gamma plays a prominent role in an interactive network of cytokines that contribute to the pathogenesis and progression of IgA nephropathy. PMID- 9327341 TI - Suppressed cell-mediated immunity and monocyte and natural killer cell activity following allogeneic immunization of women with spontaneous recurrent abortion. AB - Spontaneous recurrent abortion (SRA) has been treated by means of immunization with paternal or third-party white blood cells, yet the immunological basis for SRA and for the role of immunization protocols in pregnancy outcome remains controversial. To elucidate this question, nine women with SRA were immunized with paternal mononuclear cells and studied before and 2 weeks after immunization. Seven women who became pregnant gave birth to live newborns. Secretion of the T helper 1 cytokines IL-2 and interferon-gamma by patients, mononuclear cells decreased, while production of IL-10 increased. The levels of natural killer and lymphokine-activated killer cell-mediated cytotoxicity were markedly decreased. Monocyte functions such as secretion of IL-1 alpha, tumor necrosis factor alpha, IL-6, and cytotoxic activity decreased concurrently with elevations in IL-10 and transforming growth factor beta secretion. Production of IL-12, a pivotal regulatory cytokine, decreased. Furthermore, B7/1 expression on patients' mononuclear cells was downregulated. This resulted in a decrease in monocyte costimulatory activity of purified T cells with soluble anti-CD3, paralleled by a decline in allogeneic proliferative responses. These results suggest that the improved pregnancy success rate in women with SRA following immunization may be partly related to suppression of cell-mediated immunity and monocyte and natural killer cell activity. PMID- 9327342 TI - Numerical and functional characteristics of lymphocyte subsets in centenarians. AB - The immune system in the aged is a very interesting subject for study. In this study, analysis was extended to extrathymic T cells as well as NK cells and "conventional" T cells (i.e., thymus-derived T cells) in terms of their constitution and function in both healthy and unhealthy centenarians. Middle-aged persons were used as controls. Healthy and unhealthy centenarians showed lower levels in the proportion and absolute number of lymphocytes. The major change in the constitution of lymphocyte subsets was increased levels in the proportion of NK cells (CD56+/CD57+) and extrathymic T cells (CD3+CD57+). Inversely, conventional T cells decreased in proportion and function (i.e., proliferative response to mitogen). Although NK cells increased in centenarians, NK activity by whole lymphocytes and the purified NK fraction decreased. The difference between healthy and unhealthy centenarians was small in all parameters, the only difference being a lower level of expression of CD56 antigens on CD57+ T cells in unhealthy centenarians. These results indicate that there is a major shift in lymphocyte population from conventional T cells to NK cells and extrathymic T cells with aging. Concerning the age-associated increases in CD56+ T and CD57+ T cells, these cells correspond to NK1+ T cells in mice. PMID- 9327343 TI - Antimetabolite application: science or voodoo? PMID- 9327344 TI - The influence of age and intraocular pressure on the optic cup in a normal population. AB - PURPOSE: To determine the effects of age and intraocular pressure (IOP) on optic cup and neural rim size, and cup-disc ratio in a well defined population free from apparent glaucoma and other optic nerve disease. METHODS: Data were collected on 3654 people, 49 years or older, living in the Blue Mountains, west of Sydney. Examination included subjective refraction and Zeiss colour stereo (Carl Zeiss, Oberkochen, Germany) optic disc photographs. Magnification effects of the eye and camera were corrected. After excluding subjects with optic nerve diseases, data from 6579 normal phakic eyes of 3358 subject (91.9% of those examined) were used. RESULTS: Adjusted for optic disc size and IOP, cup diameter increased 0.01 mm, cup-disc ratio increased 0.01, and neural rim width decreased 0.01 mm for every decade of age increase. Adjusted for age and optic disc size, cup diameter increased 0.01 mm, cup-disc ratio increased 0.04, and neural rim width decreased 0.07 mm for every 10 mmHg increase in IOP. The IOP-related increase in cup-disc ratio amounted to 9.5% of the mean per 10 mmHg, while the age related increase was 1.9% of the mean. CONCLUSIONS: These data support the hypothesis that there is an age-related loss of tissue from the neuroretinal rim. However the mean change between the ages of 50 and 90 years is very small. The association between increasing IOP and smaller neural rim width could suggest a causal relationship. However, it is also plausible that IOP and optic cup size are both determined by other unmeasured factors. PMID- 9327345 TI - Probability maps of sequential glaucoma-scope images help identify significant change. AB - PURPOSE: The purpose of this study was to identify areas of the optic disc showing high variability of repeated depth measurements, and to minimize the effect of baseline variability in interpretation of possible change over time using the Glaucoma-Scope. METHODS: Seventy-four eyes from 70 subjects were analyzed with the Glaucoma-Scope. Three images were obtained on each of two separate sessions during the same day. At each location, the mean depth of the three images for each session was calculated to create a "baseline image." A contour map of standard deviation (SD) values at each topographic location was created for each subject reflecting local variability at different parts of the disc. The contour map and disc photograph were compared to determine what photographic features predicted high variability. A modified two-sample t-test was used at each topographic location to obtain p-values for the likelihood that a difference in mean depth between sessions was attributable to measurement variability alone. RESULTS: Contour plots of SD for most subject eyes showed high variability in steeply sloped areas of the disc and along large blood vessels, with low variability near the cup center. The use of probability plots for significance of depth changes between test sessions automatically accounted for increased pointwise variability. The proportion of topographic locations showing statistically significant change but attributable to chance variation when no true change has occurred approximated the predicted proportion based on our modified t-test model. CONCLUSION: A contour map of standard deviations of depth based on Glaucoma-Scope baseline images can identify areas of the disc with high variability. Statistical methods such as probability maps that account for local variability in the baseline image may be helpful in distinguishing true change from artefactual change over time. PMID- 9327346 TI - Appositional angle closure in eyes with narrow angles: comparison between the fellow eyes of acute angle-closure glaucoma and normotensive cases. AB - PURPOSE: To identify topologic features of eyes with acute primary angle-closure glaucoma before an attack as compared with those features in normotensive cases, assuming that untreated fellow eyes of acute primary angle-closure glaucoma are candidates for an acute attack. METHODS: A total of 50 eyes (12 fellow eyes of acute primary angle-closure glaucoma and 38 normotensive cases with a closure possible narrow angle) were prospectively examined by ultrasound biomicroscopy under dark-room conditions. All eyes were examined before surgical or laser intervention and had normal pupillary reaction without any topical drugs. Four predetermined angle locations per eye were examined. Parameters regarding the chamber angle configuration were measured and compared between the two groups. RESULTS: Appositional angle closures were observed in 27 locations in fellow eyes and in 48 locations in normotensive eyes with a closure-possible narrow angle. The incidence was statistically different between the two groups (69.2% in the fellow eyes and 48% in the normotensive eyes). Appositional angle closures beginning at the entrance of the angle were more frequently detected in the fellow eye group. The distance between the iris root and the bottom of the angle was significantly different between the two groups. CONCLUSION: The fellow eyes of acute primary angle-closure glaucoma have different topologic features from normotensive narrow-angled eyes and a higher incidence of appositional closure that may predispose these eyes to an imminent acute attack. PMID- 9327347 TI - Mitomycin C primary trabeculectomy in primary glaucoma of white patients. AB - PURPOSE: To evaluate the clinical outcome of eyes which underwent primary trabeculectomy with adjunctive mitomycin C (MMC) for primary glaucoma. PATIENTS AND METHODS: A prospective analysis of 25 eyes in 23 patients who underwent primary trabeculectomy with MMC for primary glaucoma was performed. Clinical outcome measures including postoperative intraocular pressure, change in logarithm of the minimum angle of resolution (LogMAR) visual acuity, and incidence of complications were measured up to 1 year postoperatively. RESULTS: Mean intraocular pressure decreased from 26.0 +/- 4.4 mmHg preoperatively to 12.5 +/- 3.9 mmHg (p < 0.0001) 1 year postoperatively. The mean LogMAR visual acuity changed from 0.23 +/- 0.19 preoperatively to 0.23 +/- 0.20 1 year postoperatively (p = 1.0). One eye developed a temporary hypotonous maculopathy and 4 eyes progressed in cataract formation. CONCLUSION: Primary trabeculectomy with MMC in eyes with primary glaucoma showed excellent pressure reduction. There were no cases of persistent hypotonous maculopathy or bleb endophthalmitis. PMID- 9327348 TI - Clinical experience with apraclonidine 0.5%. AB - PURPOSE: This study describes our long-term experience with apraclonidine 0.5% in the treatment of chronic glaucoma in clinical practice. METHODS: A retrospective review was performed of all consecutive patients treated with apraclonidine 0.5%, specifically studying the magnitude of IOP reduction, incidence of allergic reaction, frequency of ineffectiveness, and its additivity to other anti-glaucoma medications. Patients previously treated with this agent or in whom multiple simultaneous medication changes were made were excluded. RESULTS: A total of 174 patients were included in this study and followed up to 24 months. For 38 patients (21%), the drug was found to be ineffective at some point during the study. A similar number of patients developed an allergic reaction to the medication. Intraocular pressure lowering ranged from 19 to 26% overall, and 22.5 to 29% in those responding to apraclonidine 0.5%. CONCLUSION: In this study, apraclonidine 0.5% was shown to be effective in reducing intraocular pressure, both for short-term situations and for longer periods of treatment, up to 24 months. PMID- 9327349 TI - Astrocyte responses in human optic nerve head with primary open-angle glaucoma. AB - PURPOSE: To identify and characterize astrocyte responses and reactivation in human optic nerve heads from patients with primary open-angle glaucoma. METHODS: Fifteen optic nerve heads with primary open-angle glaucoma and 13 normal controls were fixed in 4% paraformaldehyde, paraffin embedded, and stained for immunofluorescence and immunoperoxidase. The antibodies used were against glial fibrillary acidic protein (GFAP) and against neural cell adhesion molecule (N CAM). RESULTS: Two subpopulations of type 1 astrocytes exist in the normal optic nerve. Type 1A astrocytes express only glial fibrillary acidic protein and type 1B express both glial fibrillary acidic protein and neural cell adhesion molecule. These are the major cell subpopulations in the lamina cribrosa and prelaminar regions. In primary open angle glaucoma, type 1B astrocytes in the prelaminar region showed increased immunoreactivity for glial fibrillary acidic protein and neural cell adhesion molecule, and cytoplasmic enlargement with thicker and longer cytoplasmic processes. At the level of the lamina cribrosa, type 1B astrocytes appeared round and the cell bodies were no longer in the cribriform plates but located in the nerve bundles. Type 1A astrocytes were not observed in the glaucomatous optic nerve head. CONCLUSIONS: Astrocyte responses in primary open angle glaucoma may underlie cellular changes that lead to axonal damage and optic nerve head remodeling. These responses may have pathogenic significance for glaucomatous optic neuropathy. PMID- 9327350 TI - Transconjunctival mitomycin C as an adjunct to conventional glaucoma filtration surgery in rabbits. AB - PURPOSE: Subconjunctival mitomycin C has been used in glaucoma filtration surgery with success. A prospective, randomized, masked, placebo-controlled study was performed to evaluate whether single transconjunctival mitomycin C applied either preoperatively or postoperatively would enhance the success of filtration surgery in rabbits. METHODS: Two groups of 5 rabbits were studied. In Group I, a Weck-Cel sponge soaked in 0.5 mg/ml mitomycin C was applied transconjunctively for 7 minutes immediately before a full thickness filtering procedure. The other eye was treated similarly with a sponge soaked in balanced salt solution. Group II first underwent filtration surgery followed by treatment with either mitomycin-c or balanced salt solution 3 days later. Postoperative intraocular pressure, bleb status, and complications were evaluated. Treatment failure was defined as postoperative pressure within 4 mmHg of that determined preoperatively or the absence of bleb formation. RESULTS: In Group I, mean time to failure (+/- SD) was significantly longer (p = 0.03) in experimental eyes (30 +/- 15.1 days) than control eyes (8.6 +/- 0.8 days). In Group II, the time to failure was 12.4 (+/- 2.6) days versus 9.6 (+/- 2.5) days in the experimental and control eyes respectively, but this was not statistically significant (p = 0.10). Transient limbal vascularization and corneal haze were seen in all experimental eyes. Serious complications included late bleb rupture in eyes pretreated with mitomycin C (all eyes in Group I) and corneal decompensation (one mitomycin-c eye). CONCLUSIONS: This study demonstrates that a single preoperative tranconjunctival application of mitomycin-c is more effective at the time of surgery than an application applied in the intermediate postoperative period. Additional studies are needed, however, to further refine both the dose and timing of mitomycin-c application during filtration surgery. PMID- 9327352 TI - The value of electrophysiological testing in glaucomatous diseases. PMID- 9327351 TI - Evaluation of optic nerve head circulation: review of the methods used. PMID- 9327353 TI - Ophthalmic generic drug approval process: implications for efficacy and safety. PMID- 9327354 TI - Activity-based costing in the operating room at Valley View Hospital. AB - This article presents an example of how one hospital reports the results of activity-based costing (ABC). It examines the composition and supporting assumptions of an ABC report for a particular procedure in the operating room (OR). It describes management uses of the information generated. It comments upon how the continuous quality improvement (CQI) is synchronized with the ABC reporting. PMID- 9327355 TI - Total quality management issues in managed care. AB - The implementation of total quality management (TQM) in health care has gone on in parallel with the growth of managed care. What is the interaction between the two? Key issues are the ascendance of cost control over quality in many areas, erosion of employee commitment and loyalty, and a short-run orientation. Associated with this is an emphasis on organizational learning rather than learning by autonomous professionals. Both TQM and managed care acknowledge the dynamic nature of clinical processes and the ability and responsibility of both institutions and clinicians to improve their processes. Both are consistent with efforts to identify and implement best practices. However, these similarities should not mask fundamental differences. Continuous improvement must shift its focus from avoiding unnecessary variation to facilitating rapid organizational learning and institutionalizing mass customization into the delivery of health services. PMID- 9327356 TI - Improving quality while managing costs in emergency medicine. AB - This article correlates quality of care with cost of care. The authors describe their experience in developing an internal measure of quality and two surrogates for cost. They examine archival data for 3,671 patients in the emergency department of a large community teaching hospital. Their results indicate statistically significant differences among emergent, urgent, and routine care assessments by triage staff, nurses, and physicians. Only 56 percent of the assessments were consistent. Triage was significantly less predictive of nursing acuity assessments than physician resource-based relative value scale codes. The authors conclude that by reducing process variation in patient acuity assessments, health care managers can improve quality of care while managing costs. PMID- 9327357 TI - The relationship among cost, quality, and competition: an analysis of obstetrics services in Missouri hospitals. AB - An understanding of the relationship among cost, quality, and competition is vital to ongoing efforts of market-based health care reform. The objectives of this study were to introduce a distance-based operational definition of competition and to examine the relationships among competition, cost, and quality within the singular product and geographic market of obstetrics services at hospitals within the state of Missouri. Correlational results indicate that increased competition is related to both increases in quality of care and costs- the characteristics of a price-insensitive market. This has obvious implications on health policy debates focusing on enhancing market competition as an avenue for health care reform. PMID- 9327359 TI - A health statistic for length of stay by reporting period. AB - Managers of long-term care organizations lack usable information about patterns of stay, because the time spent by residents typically exceeds the length of reporting and reimbursement periods. Formulas for estimating lengths of stay often yield unstable estimates of long-term stays. Consequently, little insight is gained into changes that may be occurring in stay patterns as the organization adapts its service processes to shifts in markets. An alternative length of stay statistic is proposed from which patterns of stay can be more clearly discerned. Examples utilizing both simulated and field data are presented to highlight the advantages associated with the new statistic. PMID- 9327360 TI - Health care financial and quality measures: international call for a "balanced scorecard" approach. PMID- 9327361 TI - The dangerous financial politics of AIDS "cocktail" therapies. AB - Political pressures by local AIDS activists are forcing some communities to redirect limited financial resources exclusively toward new drug therapies called protease inhibitors. These expensive and often reckless decisions threaten both the financial integrity of AIDS service organizations and the health of the public. The dangerous financial politics surrounding access to the new and complicated drug therapies for AIDS threaten to bankrupt local AIDS health service, create inescapable financial and legal liabilities, and may contribute to a worsening of the AIDS epidemic, causing the elimination of essential systems of prevention and care for the highest risk populations-adolescents and minority women. PMID- 9327362 TI - The economic impact of high-risk pregnancies. AB - To compare the costs of prenatal care, labor and delivery, and postnatal care of 775 high-risk (HR) pregnancies with costs of 2,825 low-risk pregnancies, data were collected from retrospective chart review and computerized financial records of infants and mothers. Claims paid to providers, hospitals, and ancillary services were the direct medical costs of care for Sentara Health Plan. The total prenatal, labor and delivery, and postnatal costs were more than 6 million dollars and 3.5 million dollars for premature and term babies, respectively. Postnatal and total costs were related inversely to gestational ages and birth weights and directly related to length of stay. The data indicate the substantially increased cost of identified HR pregnancies. The gestational age and birth weight correlate with postnatal and total costs. PMID- 9327363 TI - Capacity planning for the future. AB - Managed care is changing the way health care organizations plan for their futures. Traditional planning takes into account history and geography, while the new approach factors in the impact of managed care of future utilization. The new approach also incorporates strategic planning into an organization's broader strategic plan and budgeting process. The result is a more comprehensive planning method that is critical for health care organization's success. PMID- 9327364 TI - In situ Rantes and interferon-gamma gene expression in pediatric small bowel Crohn's disease. AB - BACKGROUND: Rantes (regulated upon activation, normal T cell expressed and secreted) is a chemotactic cytokine for memory T lymphocytes, monocytes, and eosinophils. The cytokine interferon-gamma (IFN-gamma) plays a key in the immune response. Their distributions and possible roles in the selective accumulation of inflammatory cells in Crohn's disease (CD) were examined by determining the expression of Rantes and IFN-gamma genes in patients with CD using in situ hybridization (ISH) on frozen and paraffin-embedded tissue sections. METHODS: Intestinal and mesenteric lymph node samples from 9 children who had undergone ileal resection for CD were examined for the presence of epithelioid-giant cell granulomas (EGCG) and Rantes and IFN-gamma messenger RNA (mRNA). Normal pediatric intestine (n = 5) and lymph nodes (n = 2) served as controls. RESULTS: Many cells in all CD specimens in the epithelial compartment, lamina propria, and the EGCG gave positive signal with the Rantes antisense probe. Labelled cells were identified on paraffin sections as lymphocytes, macrophages, and epithelioid cells. There were Rantes-positive cells in the control intestinal tissues, but many Rantes-positive cells in control lymph nodes that showed follicular hyperplasia. IFN-gamma-positive cells were present in all CD ileal and lymph node specimens, predominantly in close contact with EGCC. No positive signal was obtained with the Rantes and IFN-gamma sense control probes. CONCLUSIONS: These findings suggest that Rantes and IFN-gamma contribute to the selective accumulation of macrophages and memory T helper lymphocytes inside the granulomas and inflammatory infiltrates that are characteristic of CD. PMID- 9327365 TI - Is a gastric drainage procedure necessary at the time of antireflux surgery? AB - BACKGROUND: Gastroesophageal reflux is part of a generalized foregut motility disorder, which may also include delayed gastric emptying. With persistence of gastroesophageal reflux, or the presence of complications, including recurrent aspiration syndrome and esophageal stricture formation, surgical correction may be indicated. It is uncertain whether a procedure to resolve delayed gastric emptying is indicated at this time as well. METHODS: Sixty-seven children with proven gastroesophageal reflux had preoperative gastric emptying assessed using 99Technetium-Sn-colloid labelled milk. Delayed gastric emptying was defined as a gastric residual activity of more than 40% at 2 hours after feeding. The antireflux operation was a partial anterior fundoplication. Postoperative milk scans assessed the effect of surgery on gastric emptying. RESULTS: Gastric emptying at 2 hours improved overall from a median of 22% before surgery to 17% after surgery. In 17 patients delayed gastric emptying was identified before surgery; in 15 of those it returned to within normal limits after surgery. In 50 children with normal gastric emptying before surgery (gastric residual activity at 2 hours 16%), 14 (28%) showed delayed gastric emptying in the postoperative scan. CONCLUSIONS: Delayed gastric emptying is common in children who undergo surgery for gastroesophageal reflux disease. A partial anterior fundoplication antireflux operation improves gastric emptying to within normal limits in the majority (88%) in this group, rendering a synchronous gastric drainage procedure unnecessary. PMID- 9327366 TI - Acceptability, safety, and digestibility of spray-dried bovine serum added to diets of recovering malnourished children. AB - BACKGROUND: Specially collected, spray-dried bovine and porcine blood plasma have been incorporated previously in feeds of weanling farm animals, resulting in increased dietary intakes and greater rates of weight gain than observed in control animals. Before conducting similar trials in human populations, preliminary studies have been completed to assess the acceptability, safety, and digestibility of processed animal plasma in young children. METHODS: Masked study diets were provided sequentially to each of ten young, Peruvian children recovering from severe protein-energy malnutrition during three randomly ordered 7-day dietary periods. The control diet was prepared from rice, milk, vegetable oil, and sugar; the two study diets included spray-dried, bovine serum concentrate to replace either 25% or 50% of the milk protein of the control diet. Urine and feces were collected quantitatively during the last four days of each diet period to assess stool weight, apparent absorption of macronutrients, and retention of nitrogen. RESULTS: All children consumed the entire amounts offered of each of the diets. The mean number of daily bowel movements and mean apparent absorption and retention of nitrogen and mean apparent absorption of carbohydrate were similar for each diet. Fractional absorption of dietary lipid and of total energy increased significantly in relation to the amount of bovine serum concentrate in the diet, although this might be explained by the simultaneous replacement of milk fat with additional vegetable oil. CONCLUSIONS: Each of the diets was well accepted by the study children, and there was no evidence of any adverse effects of bovine serum concentrate. PMID- 9327367 TI - The effect of starvation on brain carnitine concentration in neonatal rats. AB - BACKGROUND: We examined carnitine concentrations in fasted neonatal rat brain to evaluate the effect of starvation on fatty acid metabolism. METHODS: The free- and acylcarnitine concentrations in neonatal rat brain and heart were determined after a 72-hour starvation period from the 3rd to 6th postnatal day. They were also determined in rats at 3 and 6 days of age fed normally by the mother rats as controls. RESULTS: In the brain, the mean free carnitine concentration in the fasted group showed an increase similar to that in normal rats and there was no difference between the fasted and 6-day-old control rats. However, the mean acylcarnitine concentration was significantly higher in the fasted group than in the control group at both 3 and 6 days of age. Almost all of the increased acylcarnitine in the fasted group was short-chain acylcarnitine. In the heart, there was no difference in the mean free carnitine concentration between the fasted group and control group at 6 days of age. The 6-day-old rats in both the fasted and control groups showed higher levels compared to 3-day-old rats in the control group. The mean acylcarnitine concentration in the fasted group was not different from that in control group at 6 days of age, while the amount of short chain acylcarnitine was less than that in the control group at 6 days of age. CONCLUSIONS: These findings suggest that in the brain, carnitine is accumulated as a result of redistribution during starvation, and is utilized for energy supply by fatty acid oxidation. PMID- 9327368 TI - Simultaneous determination of fecal fat, nitrogen, and water by near-infrared reflectance spectroscopy. AB - BACKGROUND: Determinations of fecal fat and nitrogen reveal evidence of malabsorption and assist in estimating the efficacy of pancreatic enzyme treatment. Seventy-two-hour stool collection, with chemical analysis of fecal fat, and Kjeldahl's method for measurement of fecal nitrogen are generally accepted as standard methods for making these determinations. However, these traditional methods are expensive, time-consuming, and cumbersome. This study evaluated the efficiency and validity of an alternative method, using near infrared reflectance spectroscopy (NIRS) and compared results with those of the standard methods. METHODS: Near-infrared reflectance spectroscopy is a secondary method: The instrument first has to be calibrated with samples analyzed by the standard method. Sixty-three stool samples with known fat content (range 4.79 292.5 mg/g), 24 samples with known nitrogen content (range 5.36-19.38 mg/g), and 24 samples with known water concentration (range 60.1-82.22%) served for calibration. A further 69 samples were analyzed to validate the procedure. RESULTS: There was a satisfactory correlation between the measurements produced by near-infrared reflectance spectroscopy and those produced by standard methods: fat r = 0.97; nitrogen r = 0.94; water r = 0.96. CONCLUSIONS: Near-infrared reflectance spectroscopy appears to be a reliable, simple, and rapid method of measuring different fecal components-as precise and accurate as the standard methods. Stool samples should be analyzed immediately after collecting or stored only a few days before analyzing. PMID- 9327369 TI - Colonic structural and ion transport abnormalities in suckling rabbits infected with Escherichia coli K12. AB - BACKGROUND: Escherichia coli K12 is a laboratory strain considered nonpathogenic. The purpose of this study was to examine the effect of E. coli K12 infection on colonic structure and function. METHODS: Suckling rabbits were infected at 10 days of age with 6 x 10(9) CFU E. coli by intragastric inoculation and were examined 4 to 5 days later. Segments of ileum and proximal and distal colon were removed for light and electron microscopy, and NaCl transport was examined in vitro under short-circuited conditions in Ussing chambers. RESULTS: Infection did not cause weight loss or diarrhea. Colonic mucosa was inflamed with infiltration by polymorphonuclear neutrophils mainly in the lamina propria. The proximal and distal colon exhibited reduced Na+ absorption. The proximal colon also showed increased Cl- secretion; the ileum was unaffected. CONCLUSIONS: Infection with E. coli K12 disrupts the epithelium and alters ion transport in the colon, probably as a result of mucosal inflammation. The changes indicate that nonpathogenic E. coli have the potential to cause intestinal disease. PMID- 9327370 TI - Essential fatty acids and their trans geometrical isomers in powdered and liquid infant formulas sold in Canada. AB - BACKGROUND: Animal and human studies have suggested that trans fatty acids might alter some physiological functions and adversely affect the growth and essential fatty acid balance of infants. In this context it is important to know the fatty acid composition, including the levels of trans isomers of oleic, linoleic and alpha-linolenic acids in infant formulas. METHODS: Ten liquid and fourteen powdered formulas for term infants were purchased from retail stores in Canada. The fatty acid composition of each formula was determined by capillary gas-liquid chromatography. RESULTS: All the formulas met the minimum content of 500 mg of linoleic acid/100 kcal formula (equivalent to 4.5% of energy) specified under current Canadian regulations. The formulas all met the minimum energy levels of 3% as linoleic acid and 0.7% as alpha-linolenic acid recommended recently by an ad hoc committee of Health Canada. However, in nine formulas, the proportion of linoleic acid was more than 20% of total fatty acids, and consequently, in five of them, the ratio of linoleic acid to alpha-linolenic acid exceeded the maximum ratio of 16:1 recommended by the ad hoc committee. Trans fatty acids were present in all the samples, and generally the liquid formulas displayed a higher total trans content (mean 1.9%, range 0.9-3.1%) than powdered formulas (mean 1.4%, range 0.6-2.5%). The amounts of trans isomers of linoleic and alpha-linolenic acids and the degree of isomerization of these fatty acids were also higher in liquid formulas than in powdered formulas. CONCLUSIONS: A few of the Canadian infant formulas would provide one-third of alpha-linolenic acid as trans geometric isomers. PMID- 9327371 TI - Possible antioxidant effect of vitamin A supplementation in premature infants. AB - BACKGROUND: Increased lipid peroxidation caused by oxygen free radicals is thought to be one of the common pathogenetic mechanisms for the so-called oxygen radical diseases of prematurity. Since in vitro studies have shown that various forms of vitamin A can exert antioxidant effects that are more potent than those of vitamin E (treatment with which has been ineffective in these diseases), the purpose of this prospective, controlled study was to determine whether administration of supplemental vitamin A to premature infants deficient in this vitamin would have an antioxidant effect in vivo. METHODS: Fourteen infants (1181 +/- 35 g; gestational age 29 +/- 0.04 weeks) with a serum retinol concentration at 7 +/- 2 days of age in the deficient range, lower than 0.7 mumol/l (< 20 micrograms/dl), were enrolled in the study. Infants were randomized to receive the standard amount of vitamin A or standard plus supplemental (2.6 mumol/l [2500 IU] orally each day) vitamin A, beginning at 1 week of age. Antioxidant effects of supplementation were assessed by a decrease in lipid peroxidation, quantified by the ethane content of expired air. RESULTS: Three weeks after study enrollment, total daily vitamin A intake in the infants receiving supplements was 4.565 +/- 0.236 mumol (4354 +/- 225 IU) versus 1.879 +/- 0.317 mumol/l (1792 +/- 302 IU) in infants receiving standard amounts of the vitamin. In spite of the difference in intake of vitamin A, 3 weeks after study enrollment, serum retinol concentrations did not differ between the infants given supplements and those receiving standard amounts of vitamin A, 0.70 +/- 0.21 versus 0.66 +/- 0.07 mumol/l (20 +/- 6 micrograms/dl versus 19 +/- 2 micrograms/dl, respectively). In the infants receiving supplemental vitamin A, breath ethane values declined from baseline values. There was an inverse correlation between the number of weeks of supplementation and breath ethane values, whereas there was no significant correlation between the duration of the study and breath ethane values in the infants not given supplements. CONCLUSIONS: Our data suggest that supplementation with vitamin A in a small group of vitamin A-deficient preterm infants was associated with an antioxidant effect. Although no immediate clinical benefits were associated with supplementation, the data provide the rationale for future investigations of possible antioxidant effects of (larger amounts?) of vitamin A in higher risk premature infants born with subnormal serum retinol concentrations. PMID- 9327372 TI - Celiac disease: an uncommon cause of recurrent intussusception. PMID- 9327373 TI - Hepatic vein thrombosis (Budd-Chiari syndrome) in an adolescent with ulcerative colitis. PMID- 9327374 TI - Transjugular intrahepatic portosystemic shunt placement in a child complicated by perforated Roux-en-Y portoenterostomy. PMID- 9327375 TI - Duodenal hematoma as a complication of endoscopic biopsy in pediatric bone marrow transplant recipients. PMID- 9327376 TI - Ulcers of the rectal mucosa caused by suppositories containing acetylsalicylic acid. PMID- 9327377 TI - Desmin-rich smooth muscle bundles in chronic intestinal pseudo-obstruction. PMID- 9327378 TI - Massive pericardial effusion in a child following the administration of mesalamine. PMID- 9327379 TI - Nitric oxide and the intestinal epithelial barrier: does it protect or damage the gut? PMID- 9327380 TI - Helicobacter pylori gastritis and sideropenic refractory anemia. PMID- 9327381 TI - Inhibitory effects of fluorescein isothiocyanate photoactivation on lymphatic pump activity. AB - The effects of photoactivation of fluorescein 5'-isothiocyanate (FITC)-dextran on lymphatic pump activity of rat mesenteric collecting vessel were studied in vivo. Rats were anesthetized with intraperitoneal alpha-chloralose and urethane, and the mesenteries were studied by using intravital videomicroscopic techniques. The diameter of the collecting lymph vessels were continuously monitored and lymphatic pump parameters (end diastolic diameter, end systolic diameter, stroke volume index, ejection fraction, contraction frequency, and pump flow index) were calculated. FITC-dextran (42 nmol/100 g body wt) without illumination caused no disturbance of lymphatic pump activity. Photoactivated FITC-dextran significantly increased end systolic diameter and decreased stroke volume index, ejection fraction, contraction frequency, and pump flow index. End diastolic diameter was not changed throughout the experiment. Superoxide dismutase (120 U/ml) and catalase (5000 U/ml) had no protective effect on photoactivated FITC-induced pump dysfunction, while histidine (singlet oxygen quencher, 10 mM) significantly prevented the disturbance of pump parameters. These results indicate that photoactivation of FITC induces negative chronotropic and negative inotropic effects in lymphatic pump activity through generation of singlet oxygen in the mesentery. PMID- 9327382 TI - Human omental microvascular endothelial and mesothelial cells: characterization of two distinct mesodermally derived epithelial cells. AB - Human omental microvascular endothelial (HOME) and mesothelial (MESO) cells share many phenotypic properties, but can be characterized from one another based upon a comprehensive panel of endothelial and mesothelial markers. Traditional cell markers such as von-Willebrand factor, DiI-Ac-LDL, and Ulex europaeus I lectin are not sufficient to distinguish between HOME and MESO cells. Furthermore, immunoreactivity to a panel of endothelial cell-specific monoclonal antibodies, including representatives from the known clusters of differentiation (CD), indicate that some of these antigens are coexpressed in HOME and MESO cells. In distinguishing between the two cell types, HOME and not MESO cells express E selectin, E/P-selectin, P-selectin (CD62), Le-y, and VLA-6 (CDw49f*). Moreover, HOME cells and not MESO cells form tube-like structures when cultured on Matrigel. MESO cells differ from HOME cells based upon (1) the expression of cytokeratins; (2) their rapid proliferation in response to platelet-derived growth factor; and (3) a change from an epitheliod to fibroblast-like morphology in response to tumor necrosis factor and epidermal growth factor. Both HOME and MESO cells express tissue plasminogen activator and plasminogen activator inhibitor, but urokinase activity is only expressed by MESO cells. As there is no one universal endothelial or mesothelial cell marker that can specifically confirm the identity of these cells, it appears necessary to employ a comprehensive panel of cell markers to rule out the possibility of misidentifying a cell culture. PMID- 9327383 TI - Two-stage isolation procedure for obtaining homogenous populations of microvascular endothelial and mesothelial cells from human omentum. AB - The human omentum is a highly vascularized tissue often advocated as a source of human microvascular endothelial (HOME) cells. The omentum also contains mesothelial (MESO) cells and isolation protocols published to date do not describe a separation of the two cell populations. Using a two-stage collagenase digestion procedure, homogenous populations of HOME and MESO cells are obtained from the same omental tissue sample. HOME and MESO cells are both simple squamous epithelial cells and consequently are often difficult to discriminate between based on morphology and reactivity with many of the conventional endothelial and mesothelial cell markers. Both HOME and MESO cells form typical cobblestone, contact-inhibited monolayers, metabolize DiI-Ac-LDL, and are immunoreactive to von Willebrand Factor and Ulex europeaus I lectin. However, MESO cells are distinguishable from HOME cells based upon their expression of cytokeratins. Moreover, HOME cells and not MESO cells form capillary-like structures when cultured on Matrigel. It appears that HOME and MESO cells share many phenotypic properties, but are distinguishable from one another based upon a comprehensive panel of endothelial and mesothelial markers. Both cell types should be useful for studying the biology and pathology of the human microvasculature in vitro. PMID- 9327384 TI - Constitutive and stimulated expression of ICAM-1 protein on pulmonary endothelial cells in vivo. AB - The expression of intercellular adhesion molecule-1 (ICAM-1) on pulmonary endothelial cells after stimulus and subsequent binding of neutrophils is a first step leading to lung injury. A similar process may dictate the binding of tumor cells to the pulmonary endothelium during metastasis. We report the development of a new technique that allowed us to monitor the location and relative expression of ICAM-1 levels on the luminal surface of the pulmonary microvasculature in vivo. This technique uses intravital microscopy together with a two-step labeling procedure involving fluorescent microspheres. Constitutive expression of ICAM-1 was not detectable to a significant level by our model, but expression was observed after upregulation by the systemic administration of TNF alpha. Sprague-Dawley rats were injected with 0-5.0 micrograms/kg TNF alpha and ICAM-1 expression was monitored through 24 hr. ICAM-1 expression was related to both the dose of TNF alpha administered and the time elapsed between injection of TNF alpha and observation. Injection of 5 micrograms/kg TNF alpha caused upregulation of ICAM-1 protein expression from 0.30 +/- 2.76 binding events/175,000 microns2 to 62.6 +/- 5.48 through 4 hr observation, after which levels returned to near baseline within 24 hr. The delay required for maximal expression is likely related to the time required for the cell to respond to the stimulus and generate ICAM-1 protein. Reductions in the relative numbers of ICAM 1 protein expressed between 4 and 24 hr in vivo are likely a result of protein turnover after the initial stimulus. PMID- 9327385 TI - Endothelial ectoenzyme assays estimate perfused capillary surface area in the dog lung. AB - Whether the pulmonary vascular bed accommodates flow-induced increases in blood volume mainly through recruitment of previously unperfused capillaries or distension of already perfused vessels remains controversial. The modified first order reaction parameter of an enzyme and substrate, Amax/K(m), is, under nontoxic conditions, proportional to enzyme mass. Thus for ACE, an endothelium bound ectoenzyme uniformly distributed along the luminal surface of the pulmonary capillary bed, Amax/K(m) is proportional to the dynamically perfused capillary surface area (PCSA). We estimated single-pass translobar hydrolysis and calculated the corresponding Amax/K(m) values of the synthetic ACE substrate 3H benzoyl-Phe-Ala-Pro (BPAP), under first-order reaction conditions, in isolated blood-perfused dog lung lobes. We additionally studied blood flow distribution using radioactive microsphere techniques. Experiments were performed under zone III conditions over a wide range of lobar blood flow rates (Qb). As Qb was increased, Amax/K(m) rose linearly, while lobar vascular resistance (LVR) decreased, suggesting capillary recruitment rather than distension. Single pass BPAP hydrolysis (v approximately 2.9 at resting Qb) was not altered over a wide range of Qb, indicative of unchanging capillary transit times. When full capillary recruitment was achieved (at Qb > 70 ml/min/g lung wet weight), further Qb elevations failed to increase Amax/K(m), but decreased BPAP hydrolysis, denoting shorter transit times through the fully recruited capillary bed. Our data indicate that, as previously shown for rabbit lung, in this canine model, increases in pulmonary blood volume are mainly accommodated through recruitment of previously unperfused capillaries throughout the entire lung. PMID- 9327386 TI - Quantification of the initial lymphatic network in normal human forearm skin using fluorescence microlymphography and stereological methods. AB - Fluorescence microlymphography enables the visualization of the initial lymphatic network in human dermis in vivo. In order to determine which objective, measurable parameters are likely to be of value in future studies, microlymphangiograms were obtained from normal forearm skin of eight men and nine women of a range of ages and analyzed stereologically to calculate: (1) lymphatic length density at a specified radial distance, r, from the site of injection of the fluorescent dye (LDr, cm-1); (2) maximum lymphatic density (LDmax, cm-1); (3) total length of filled lymphatic (LL, cm); (4) maximum radial spread of vessels (mm). There was no significant difference between the right and left arms for peak LDr, LDmax, LL, or spread. Mean values (+/-SD) for the right and left arms combined (n = 24) were 13.34 +/- 7.61 cm-1; 19.31 +/- 8.96 cm-1; 12.04 +/- 10.04 cm; and 7.58 +/- 3.30 mm, respectively. Unexpectedly, three of the four parameters were significantly lower in women compared to men. For women LDmax was 30% lower than for men (P = 0.04), LL was 59% lower (P = 0.032), and spread was 39% lower (P = 0.007). Peak LDr did not differ significantly. Stereological analysis of fluorescent microlymphangiograms enables quantification of the dermal initial lymphatic network and hence the objective study of the influence of gender, age, and disease (including treatment interventions). PMID- 9327387 TI - High-resolution microscopic determination of hepatic NADH fluorescence for in vivo monitoring of tissue oxygenation during hemorrhagic shock and resuscitation. AB - Impaired microvascular oxygen supply reduces oxidative phosphorylation and causes an increase in cellular NADH, which was monitored densitometrically in vivo by high-resolution fluorescence microscopy (330-390/ > 430 nm excitation/emission wavelengths) in rat livers (n = 8) subjected to hemorrhagic shock and resuscitation. At each time point, NADH fluorescence was recorded from 10 different observation fields of the left liver lobe. Withdrawal of a total of 4.5 ml arterial blood for induction of volume-controlled hemorrhagic shock resulted in an increase in NADH fluorescence by approximately 31% from 45.1 +/- 3.9 to 59.2 +/- 4.2 aU, which was associated with a fall of arterial blood pressure from 110 +/- 3 to 51 +/- 8 mmHg, a decrease in hepatic tissue oxygenation (flexible polarographic surface electrode) from 18 +/- 2 to 2 +/- 1 mmHg, and a restriction of hepatic bile flow from 1.7 +/- 0.1 to 0.5 +/- 0.2 microliter/min x g. Normovolemic resuscitation with 10% hydroxyethylstarch failed to completely restore the metabolic state of liver tissue (NADH fluorescence 49.9 +/- 3.1 aU), arterial blood pressure (83 +/- 8 mmHg), hepatic tissue oxygenation (7.4 +/- 1.5 mmHg), and hepatocellular excretory function (1.3 +/- 0.1 microliters/min x g). During both shock and resuscitation, the ratio between pericentral and periportal NADH fluorescence intensities slightly increased, but calculation of coefficients of variance of interlobular NADH fluorescence did not reveal an increase in heterogeneity of tissue metabolic state. Significant correlations were found between NADH fluorescence and both hepatic tissue oxygenation (r2 = 0.78, P < 0.01) and hepatic bile flow (r2 = 0.85, P < 0.01), indicating that high resolution intravital microscopic assessment of NADH fluorescence reflects appropriately the relation between local oxygen supply and demand in hepatic tissue in vivo. PMID- 9327388 TI - Effects of needle insertion in tumors on interstitial fluid pressure. PMID- 9327389 TI - Inhibition of angiogenesis by oral ingestion of powdered shark cartilage in a rat model. PMID- 9327390 TI - The influence of hypnoid relaxation on the hypothalamic control of thermoregulatory cutaneous blood flow. PMID- 9327391 TI - Reduced leukocyte adhesion response and absence of slow leukocyte rolling in interleukin-8 receptor-deficient mice. PMID- 9327392 TI - A novel method for the purification of recombinant subunit I of the Dolichos biflorus seed lectin. AB - Lectins are carbohydrate-binding proteins that are ubiquitous in nature. Their ability to specifically bind carbohydrates has been used as a means of purification mainly through affinity chromatography techniques. Plant lectins are one of the most thoroughly studied class of lectins, however, details of their in situ function remains elusive. Recent advances in recombinant DNA techniques have been used in several laboratories to study the function of these lectins by heterologous overexpression. The larger subunit of the Dolichos biflorus seed lectin was described by Chao et al. in 1994 and purification through affinity chromatography techniques was described. Here we report on a new method for the purification of this recombinant protein with techniques that are not dependent on the ability of the lectin to bind sugars. This method may have uses in the purification of mutant proteins that may not bind carbohydrates. Characterization of the purified protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization (MALDI) mass spectroscopy shows that the lectin is over 99% pure with a molecular weight of 27,090 +/- 16.17 Da, and hemagglutination assays confirm that the lectin retains its biological activity. PMID- 9327393 TI - Subtractive hybridization of cDNA from small amounts of plant tissue. AB - A subtractive hybridization method is described that allows the generation of a subtractive gene library from small amounts of plant or other eukaryotic tissues. The method uses paramagnetic oligo-dT beads to capture poly-adenylated mRNA and to synthesize the complementary cDNA on a solid support. The use of magnetic beads facilitates the change of reaction buffers and the removal of primers and minimizes yield losses. Subtracted material obtained from this method can either be cloned directly or used to screen a specific library. PMID- 9327394 TI - Rapid PCR amplification of chimeric products and its direct application to in vivo [corrected] testing of recombinant DNA construction strategies. AB - This article describes a simple and rapid method for efficient production of chimeric products by polymerase chain reaction (PCR). This protocol is amenable to site-directed mutagenesis strategies and can be done without the time consuming gel purification step. The PCR products generated can also be directly used for direct gene transfer into plant cells without further subcloning to test construction strategies. PMID- 9327397 TI - Molecular cloning of PCR fragments with cohesive ends. AB - Use of the polymerase chain reaction (PCR) provides a convenient means of generating DNA fragments for insertion into plasmids. Large quantities of the desired insert, bounded by convenient restriction sites, may be synthesized. The primers are chosen to span a known region of interest, and extended at their 5' ends to include the desired restriction sites. Amplification of the target sequence is followed by precipitation of the product with ammonium acetate and ethanol to remove the primers. A small amount of product is analyzed by gel electrophoresis to ensure correct amplification, the remainder is digested with the appropriate restriction enzyme(s). Restricted insert DNA is added to similarly restricted plasmid DNA in several ratios and incubated with DNA ligase to recircularize. Ligation products are used to transform competent bacteria. Clones containing inserts are identified by restriction digestion of plasmid minipreps from bacterial colonies. PMID- 9327396 TI - Methods for the analysis of DNA-protein interactions. AB - The interaction of proteins with DNA is a central theme of molecular biology. In this article, we review some of the principal techniques currently used for the identification and characterization of DNA binding proteins, and for investigation of the molecular interactions that are responsible for the recognition of specific DNA sequences. PMID- 9327395 TI - Antiviral ribozymes. New jobs for ancient molecules. AB - Catalytic RNAs are a genetic property not only of some particular viroids or viruses, but also are more common naturally among eukaryotes and even prokaryotes than earlier expected. However, the major interest in ribozymes results from their potential for development of "tailor-made" cDNA constructions designed to be transcribed into catalytic RNAs that will recognize by hybridization and destroy by specific cleavage their cellular or viral RNA targets. The efficiency of an antiviral ribozyme is determined by both the accessibility and sequence conservation of the target region, as well as the design of the ribozyme: its type, size, and composition of flanking sequences; expression rates; and cellular compartment localization. Until now the most frequently selected viral target is the human immunodeficiency virus, where an up to a 10(4)-fold inhibition in its progeny production has been achieved. Although the first generation ribozymes focused on improvements in basic design and expression rates, more recently the efficiency of antiviral catalytic activity has been increased by employing polyribozymes and/or multitarget ribozymes, as well as special constructions to enhance the cellular co-compartmentation of the ribozyme with its viral RNA target. PMID- 9327399 TI - In vitro transcription systems from cultured cells and fat-body tissue of the silkworm, Bombyx mori. AB - This article describes the development of cell-free transcription systems from the cultured cells and fat body tissues of a Lepidpteran insect, the silkworm, Bombyx mori. Detailed protocols are provided for the culture of a B. mori cell line, rearing larvae, preparation of whole cell as well as nuclear extract and conditions for in vitro transcription of cloned plasma protein gene templates. PMID- 9327398 TI - Reverse transcriptase. The use of cloned Moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA. AB - Reverse transcriptase (RT) is the key enzyme required for conversion of RNA to DNA. Cloning of Moloney murine leukemia virus (MMLV) RT has enable engineering an RT that lacks endogenous RNase H activity. RT catalyzes cDNA synthesis more efficiently in the absence of RNase H. We describe here a number of properties of MMLV RT and RNase H-minus MMLV RT not summarized in a single location elsewhere, providing a basis for best use of these enzymes in cDNA synthesis. In addition, general guidelines and detailed protocols are provided for use of MMLV RTs in one tube double-stranded cDNA synthesis, in [32P]cDNA synthesis, and in RT-PCR and long RT-PCR. PMID- 9327400 TI - Disturbances of neuromuscular interaction may contribute to muscle weakness in spinal muscular atrophy. AB - During a critical period of development, motoneurones and muscles are dependent on functional interaction with each other for their survival. In certain neuromuscular disorders such as spinal muscular atrophy (SMA), motoneurones die and muscle development is seriously affected. Recently advances have been made towards understanding the genetic basis of this disease, and a particular gene, i.e. the survival motoneurone gene (SMN), has been identified and found missing in SMA patients. Nevertheless the function of this gene is not clear, it may be involved in the control of the development of either the motoneurone or the muscle. Here we report that disrupting neuromuscular interaction during the early postnatal period has similar consequences on the development of muscles and motoneurones to those seen in patients with SMA, in that there is a loss of motoneurones and muscle function is severely impaired. In view of this, we discuss the possibility that these symptoms in SMA patients may be due to a disturbance of neuromuscular interaction during a critical stage of development. PMID- 9327401 TI - Changes of laminin beta 2 chain expression in congenital muscular dystrophy. AB - We studied the distribution of laminin beta 2 chain in the skeletal muscle basement membrane of 16 patients with congenital muscular dystrophy (CMD) by immunohistochemistry. A dramatic reduction in the laminin beta 2 staining was observed in four patients with classical merosin-negative CMD. A moderate reduction of laminin beta 2 labelling was observed in four patients with partial merosin deficiency and two patients with merosin-positive CMD. Two patients with merosin-positive CMD had no apparent changes in the expression of laminin beta 2. In three patients and one fetus diagnosed as Walker-Warburg syndrome (WWS) the laminin beta 2 pattern was similar to normal controls. We conclude that a primary deficiency in the laminin alpha 2 chain may lead to a vast or moderate reduction in the laminin beta 2 chain in the skeletal muscle membrane. PMID- 9327402 TI - Oxidative stress as a potential pathogenic mechanism in an animal model of Duchenne muscular dystrophy. AB - Dystrophin-deficiency results in degeneration of most, but not all, skeletal muscles. The mechanisms responsible for degeneration of limb muscle and sparing of extraocular muscle are not known. To address the notion that muscle pathology may be free radical-mediated, we evaluated antioxidant enzyme activities and lipid peroxidation products (TBARS) content in mdx and control mice. TBARS content and the activities of total superoxide dismutase, selenium dependent glutathione peroxidase, glucose-6-phosphate dehydrogenase and catalase were consistently higher in both affected and spared muscles of mdx mice. These data suggest that oxidative stress may be constitutively present in mdx muscle, but may not be the principal pathogenic mechanism. To further test the hypothesis of oxidative stress involvement in dystrophinopathies, control strain and mdx mice were subjected to chronic hyperoxia. The pattern of antioxidant enzyme activities and TBARS content from hyperoxic control strain mice was similar to that of normoxic mdx mice, suggesting that a similar level of oxidative stress was induced. In conclusion, this study has provided indirect evidence for oxidative stress in dystrophin-deficient muscle. PMID- 9327403 TI - Association of genetically proven deficiencies of myophosphorylase and AMP deaminase: a second case of 'double trouble'. AB - We studied a 25-year-old man with paresis of the limbs and neck, scapular atrophy, facial weakness, exercise intolerance and frequent episodes of myoglobinuria. Muscle histochemistry and biochemistry revealed a combined defect of myophosphorylase and AMP deaminase. Molecular genetic analysis showed that the patient was homozygous for the two most common mutations associated with myophosphorylase and AMP deaminase deficiencies. This is the second documented case of genetic 'double trouble', which should be looked for in patients with unusual severe phenotypes. PMID- 9327404 TI - Toxoplasmic polymyositis revisited: case report and review of literature. AB - Toxoplasmosis can cause polymyositis either by reactivation or by recent infection. Inconsistent response to antiprotozoal therapy has been the strongest argument against toxoplasmic polymyositis as a separate entity. We report a biopsy-proven case of toxoplasmic polymyositis in a cardiac transplant patient presenting with a severe proximal weakness, myopathic, electromyographic changes and ten-fold increase of anti-Toxoplasma antibodies. An early antiprotozoal therapy and plasmapheresis led to recovery. A review of previously reported cases of toxoplasmic polymyositis suggests that an early antiprotozoal therapy is the most important variable affecting the outcome of this disease. We propose that toxoplasmic polymyositis has two phases: acute, responsive to antiprotozoal therapy, and chronic, manifested by altered immune response requiring steroids. We suggest that all patients presenting with polymyositis should have serological tests for toxoplasmosis as a part of their initial evaluation and an early trial of antiprotozoal therapy in case of positive findings. PMID- 9327405 TI - 42nd ENMC Sponsored International Workshop: X-linked cardiomyopathies. 21-23 June 1996, Naarden, The Netherlands. PMID- 9327406 TI - 48th ENMC International Workshop: drug trials and clinical research in ALS. 12-14 January 1997, Naarden, The Netherlands. PMID- 9327408 TI - Neuromuscular disorders: gene location. PMID- 9327407 TI - 49th ENMC-Sponsored International Workshop: fibrodysplasia (myositis) ossificans progressiva (FOP). 14-16 February 1997, Naarden, The Netherlands. PMID- 9327409 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 9327410 TI - [Receptors and development of therapeutic agents: studies based on new approaches for receptor research]. PMID- 9327411 TI - [Pharmacological approaches to the mechanism of vascular-, glial- and neuronal responses following brain ischemia]. PMID- 9327412 TI - [Central serotonin receptor subtypes and drug development]. PMID- 9327413 TI - [Regulation of cell functions using bioactive molecules that recognize and/or modify cell membrane components]. PMID- 9327414 TI - [Cardiovascular pharmacology--present and future]. PMID- 9327415 TI - [Specificity and efficacy of antagonists of autacoids or inhibitors for their biosynthesis: fundamental and clinical approaches]. PMID- 9327416 TI - [New vistas of salivary function and therapy]. PMID- 9327417 TI - [Molecular pharmacological analysis of signal transduction by synthetic compounds]. PMID- 9327418 TI - [Role of endothelin in cardiovascular regulation]. PMID- 9327419 TI - [Molecular basis underlying inflammatory damages and phenotypic changes of airway epithelial cells]. PMID- 9327420 TI - [Mechanical stress and cellular function: physiological and pharmacological points of view]. PMID- 9327422 TI - [Signal transduction and function of astrocytes]. PMID- 9327421 TI - [New animal models for the diseases of central nervous systems and screening of new drugs]. PMID- 9327423 TI - [Strategies for bioradical detection in pharmacology]. PMID- 9327424 TI - [Regulation of age-related changes and aging markers]. PMID- 9327425 TI - [Elucidation of intracellular signalling using physiologically active substances and its application to creation of excellent medicines]. PMID- 9327426 TI - [Electrophysiological basis for excitation and conduction abnormalities in atrial arrhythmias]. PMID- 9327427 TI - [Brain metal ions and regulation of neural functions]. PMID- 9327428 TI - [Cholinergic functions in CNS: current approaches and future aspects]. PMID- 9327429 TI - [Neuroactive substances and emotional responses: In special reference to neuro endocrine-immune axis]. PMID- 9327430 TI - [Physiological and protective actions of radicals on the blood and cardiovascular systems]. PMID- 9327431 TI - [Recent advances in pathology and pharmacology of vascular smooth muscle]. PMID- 9327432 TI - [Rational strategies for discovering new drugs]. PMID- 9327433 TI - The future of combinatorial chemistry as a drug discovery paradigm. PMID- 9327434 TI - Flow cytometry applications in pharmacodynamics and drug delivery. AB - We highlight recent advances in flow cytometry that have potential applications in the pharmaceutical sciences, particularly in pharmacodynamics and drug delivery. These advances are discussed in the context of the preclinical development of anticancer agents, immunosuppressants and immunomodulators, and oligonucleotides and gene therapy. PMID- 9327436 TI - A population growth model of dissolution. AB - PURPOSE: To develop a new approach for describing drug dissolution which does not require the presuppositions of time continuity and Fick's law of diffusion and which can be applied to both homogeneous and heterogeneous media. METHODS: The mass dissolved is considered to be a function of a discrete time index specifying successive "generations" (n). The recurrence equation: phi n + 1 = phi n + r(1 - phi n)(1 - phi n X0/theta) was derived for the fractions of dose dissolved phi n and phi n+1, between generations n and n + 1, where r is a dimensionless proportionality constant, X0 is the dose and theta is the amount of drug corresponding to the drug's solubility in the dissolution medium. RESULTS: The equation has two steady state solutions, phi ss = 1 when (X0/theta) < or = 1 and phi ss = theta/X0 when (X0/theta) > 1 and the usual behavior encountered in dissolution studies, i.e, a monotonic exponential increase of phi n reaching asymptotically the steady state when either r < theta/X0 < 1 or r < 1 < theta/X0. Good fits were obtained when the model equation was applied to danazol data after appropriate transformation of the time scale to "generations". The dissolution process is controlled by the two dimensionless parameters theta/X0 and r, which were found to be analogous to the fundamental parameters dose and dissolution number, respectively. The model was also used for the prediction of fraction of dose absorbed for highly permeable drugs. CONCLUSIONS: The model does not rely on diffusion principles and therefore it can be applied under both homogeneous and non-homogeneous conditions. This feature will facilitate the correlation of in vitro dissolution data obtained under homogeneous conditions and in vivo observations adhering to the heterogeneous milieu of the GI tract. PMID- 9327435 TI - Sex hormones differentially regulate isoforms of UDP-glucuronosyltransferase. AB - PURPOSE: To investigate the role of sex hormones in the regulation of UDP glucuronosyltransferase (UGT). METHODS: We examined liver from adult, prepubertal, gonadectomised and gonadectomised plus hormone replaced rats of both sexes. Immunohistochemistry and immunoblots were performed using a polyclonal UGT antibody to a number of family 1 and family 2 UGT isoforms. Northern blot analysis was performed utilising cDNA probes to family 1 and family 2 isoforms. RESULTS: Immunohistochemistry demonstrated variations in intensity and distribution of staining in the hormonally manipulated rats. Immunoblots showed variations in individual band intensity between rat groups. Immunoblots using a more specific antibody (anti-17 beta-hydroxysteroid UGT, which recognises UGT2B3 and UGT2B2) demonstrated marked differences between male and female rats and significant alterations after gonadectomy and testosterone replacement in the male rats. In northern analysis, UGT2B3 and 2B1 mRNA were significantly higher in adult males than females, and in prepubertal males compared to prepubertal females. In male rats, gonadectomy resulted in a 45-53% reduction in UGT2B3 and 2B1 levels respectively, which increased significantly with testosterone treatment to greater than normal adult levels. No change in UGT2B3 or 2B1 occurred after gonadectomy in females. In contrast, UGT1*1 mRNA tended to be higher in adult female and prepubertal female rats than in their male counterparts. In females, gonadectomy resulted in significant up-regulation of UGT1*1, while gonadectomy plus oestradiol treatment resulted in markedly reduced levels. UGT1*1 mRNA was not significantly altered by gonadectomy in males. CONCLUSIONS: This study demonstrates the differential effects of sex hormones on the expression of isoforms from the two phylogenetically distinct UGT families. PMID- 9327437 TI - Human intestinal permeability of piroxicam, propranolol, phenylalanine, and PEG 400 determined by jejunal perfusion. AB - PURPOSE: To determine the human jejunal permeabilities of compounds utilizing different transport mechanisms using a regional perfusion approach and to establish a standard procedure for determining drug permeability class to be used for the establishment of drug product bioequivalence standards. METHODS: Six healthy male volunteers participated in this study. A multi-lumen perfusion tube was inserted orally and positioned in the proximal region of the jejunum. A solution containing piroxicam, phenylalanine, propranolol, PEG 400 and PEG 4000 was perfused through the intestinal segment at a rate of 3.0 ml/min. Perfusate samples were quantitatively collected every 10 minutes for two 100 minute periods with an intermediate wash out period to determine intra and intersubject variation. RESULTS: The mean P(eff) (+/-SD) of piroxicam, phenylalanine, propranolol, and PEG 400 were 10.40 +/- 5.93, 6.67 +/- 3.42, 3.59 +/- 1.60, 0.80 0.46 x 10(-4) cm/sec, respectively. The coefficient of variation for the intersubject variability, first and second perfusion periods were: piroxicam, 60.5% and 57.1%; phenylalanine, 52.8% and 57.8%; propranolol, 62.1% and 44.6%; and PEG 400, 81.7% and 42.3%, indicating a slightly lower CV for the second perfusion period in the same subject. The intrasubject CV's between the two perfusion periods were: 19.4%, 21.3%, 23.6% and 41.0% respectively, indicating a smaller intraindividual variation for all compounds studied. CONCLUSIONS: Piroxicam, a nonpolar drug exhibited the highest permeability of the compounds studied. The intrasubject CV was lower than the intersubject CV, indicating consistent permeability estimation within subjects. The methodology is useful for permeability estimation regardless of absorption mechanism and can be used to establish a consistent data base of human permeabilities for estimation of human drug absorption and for establishing the biopharmaceutic permeability class of drugs. PMID- 9327438 TI - Pharmacokinetic model of ascorbic acid in healthy male volunteers during depletion and repletion. AB - PURPOSE: To develop a new pharmacokinetic model for ascorbic acid (vitamin C) since no previously published model describes ascorbic acid absorption and disposition over a broad physiologic range of doses and plasma concentrations. METHODS: A new model was developed through exploratory simulations. The model was fitted to pharmacokinetic data obtained from seven healthy volunteers who underwent ascorbic acid depletion then gradual repletion. Concentrations of ascorbic acid were measured in plasma and urine. Final pharmacokinetic model parameter estimates were obtained using nonlinear regression analysis. RESULTS: The new model included saturable absorption, distribution and renal tubular reabsorption parameters. The model described ascorbic acid concentrations in plasma, cells, and urine during depletion and gradual repletion phases with a residual error less than 15%. CONCLUSIONS: The model was useful for obtaining a new understanding of the likely causes for the complex concentration-time profile observed during gradual repletion. At doses of 200 to 2500 mg per day, the plateau in pre-dose concentrations is largely due to apparent saturation of tissue uptake and less a function of oral bioavailability and renal excretion than previously thought. PMID- 9327439 TI - Application of pharmacodynamic modeling for designing time-variant dosing regimens to overcome nitroglycerin tolerance in experimental heart failure. AB - PURPOSE: Prolonged continuous administration of nitroglycerin (NTG) leads to hemodynamic tolerance. We used a previously developed pharmacokinetic pharmacodynamic (PK/PD) model of NTG tolerance in experimental heart failure to test whether dosage regimens, designed from this model, may allow avoidance of tolerance development upon continuous NTG infusion. METHODS: Simulation experiments (using ADAPT II) were performed to evolve a time-variant infusion regimen that would theoretically provide sustained hemodynamic effect (30% reduction in left ventricular end-diastolic pressure, LVEDP) throughout 10 hours of drug dosing. A computer controlled infusion pump was utilized to deliver this time-variant input. Infusion experiments were then conducted in CHF rats to challenge the predictability of the applied PK/PD model. RESULTS: Simulations showed that exponentially increasing input functions provided more sustained LVEDP effects when compared to linear or hyperbolic input functions delivering the same total NTG dose. A computer-selected infusion regimen of 6.56e0.00156 x minutes micrograms/min was anticipated to provide the desired hemodynamic profile in our animal model. Experiments conducted in rats with congestive heart failure (n = 4) confirmed the prediction of sustained hemodynamic effect without tolerance (28 +/- 4% reduction in LVEDP at 10 hrs). CONCLUSIONS: These findings support the utility of our PK/PD model of NTG tolerance in predicting NTG action, and serve as an example of therapeutic optimization through PK/PD considerations. PMID- 9327440 TI - Utilization of an amorphous form of a water-soluble GPIIb/IIIa antagonist for controlled release from biodegradable microspheres. AB - PURPOSE: We prepared injectable microspheres for controlled release of TAK-029, a water-soluble GPIIb/IIIa antagonist and discussed the characteristics of controlled release from microspheres. METHODS: Copoly(dl-lactic/glycolic)acid (PLGA) microspheres were used for controlled release of TAK-029 [4-(4 amidinobenzoylglycyl)-3-methoxycarbonyl-2-oxopiperazine++ +-1-acetic acid]. They were prepared with a solid-in-oil-in-water (S/O/W) emulsion solvent evaporation technique using either a crystalline form or an amorphous form of the drug. RESULTS: An amorphous form of TAK-029 gave more homogeneous S/O dispersion and higher viscosity than its crystalline form when added to dichloromethane solution of PLGA, resulting in a high drug entrapment into microspheres and a well controlled release of the drug. Additions of sodium chloride into an external aqueous phase and L-arginine into an oil phase also increased entrapment of the drug, and reduced initial burst of the drug from the microspheres. The microspheres demonstrated a desirable plasma level profile in therapeutic range (20-100 ng/ml) for 3 weeks in rats after single subcutaneous injection. CONCLUSIONS: A well-controlled release of TAK-029, a water-soluble neutral drug, with small initial burst was achieved by utilizing its amorphous form as a result of possible interaction with PLGA and L-arginine. PMID- 9327441 TI - Effect of freezing rate on the stability of liposomes during freeze-drying and rehydration. AB - PURPOSE: In the present study we examined the effect of the freezing protocol on carboxyfluorescein (CF) retention in liposomes after freeze-drying and rehydration. METHODS: Liposomes were frozen slowly at 0.5 degree C/min, or quickly by submerging the samples in boiling nitrogen before freeze-drying. The thermal behaviour of the frozen dispersions was analysed by Modulated Temperature Differential Scanning Calorimetry (MTDSC). The dried cakes were analysed by SEM, MTDSC and FTIR. The % encapsulated CF of the (re)hydrated liposomes was determined by fluorimetry after GPC, their vesicle size was measured by the Dynamic Light scattering Technique and their bilayer transition was studied by DSC. RESULTS: Slow freezing resulted in a markedly higher CF retention after freeze-drying and rehydration as compared to quick freezing. The effect of the freezing rate depended on the lipid composition and was most pronounced for rigid liposomes. The damage caused by quick freezing did not occur after a freezing/thawing cycle. The freezing protocol did not influence the interaction between the phospholipids and the lyoprotectants (sucrose, trehalose or glucose) in the freeze-dried state. However, analysis by DSC of dipalmitoylphosphatidylcholine (DPPC): dipalmitoylphosphatidylglycerol (DPPG) = 10:1 and DPPC liposome dispersions showed that the freezing protocol affected the bilayer melting characteristics of these liposomes after freeze-drying and rehydration. CONCLUSIONS: A proper design of the freezing protocol is essential to achieve optimal stability of rigid liposomes during a freeze-drying and rehydration cycle. PMID- 9327442 TI - Theory of force transducer design optimization for die wall stress measurement during tablet compaction: optimization and validation of split-web die using finite element analysis. AB - PURPOSE: 1) To illustrate how computer aided engineering stress analysis can be used to improve the transducer design process for tablet press instrumentation; 2) to use these optimal design procedures for the geometric optimization of a cylindrical, segmented, and a novel split-web die design. Discussion includes the selection of optimal die wall thickness, segment cutting angle, strain gage placement, Wheatstone bridge configuration, and the influence of tablet height and position within the die on signal output. METHODS: Stress analysis was done with a finite element analysis (FEA) software package running on a personal computer. RESULTS: For the segmented die, the admissible range of die wall thicknesses depends upon cutting angle; the signal output is non-linear because the stress distribution in the die wall is influenced by tablet height and position within the die. For the split-web die, the optimal configuration consists of a 1/8 in. sensing web with a strain gage located at the peak of the sensing-web arch. This prototype had a linear calibration curve (r2 = 0.999) with no hysteresis. Radial versus axial stress transmission curves for: starch and sodium chloride were consistent with literature data. CONCLUSIONS: Finite element analysis (FEA) is a useful numerical tool for the systematic optimization of tablet press instrumentation. By enclosing the sensing web of a three layered die design in a cylinder, the split-web design can be directly mounted without modification of the die table. PMID- 9327443 TI - Study of hygroscopic properties of aqueous mixtures of disodium fluorescein and sodium chloride using an electrodynamic balance. AB - PURPOSE: The purposes of this study are: a) to demonstrate the use of an Electrodynamic Balance (EDB) to investigate the hygroscopic properties of pharmaceutical aerosols; and b) to evaluate the applicability of the Zdanovskii Stokes-Robinson model (ZSR) in the associated data analysis with multicomponent pharmaceutical aerosols. METHODS: The compositional dependence of the water activity of two model materials commonly employed in the study of pharmaceutical aerosols, namely, NaCl and Disodium Fluorescein (DF), was investigated using an EDB. The water contents of single levitated droplets of NaCl and DF and their mixtures at mass ratios of 1:3, 1:1, 3:1, and 6:1 from dilute concentration to high supersaturation were determined as a function of relative humidity (RH). RESULTS: At decreasing ambient RH, supersaturated aqueous NaCl droplets lose water and crystallize to form dry solid particles at an RH of approximately 50%. Aqueous DF droplet continues to lose water until it reaches a final state containing about 20% by mass of residual water. Mixed solutions of DF and NaCl crystallize at an RH of approximately 50% and then continue to lose water at lower RHs. The resulting "dried" particle still contains water whose amount depends on the mass ratios of DF and NaCl in the mixture. Good prediction of water activity of the DF-NaCl mixture can be achieved with the ZSR model. Collection of a full set of water activity-composition data at each mass ratio of DF-NaCl requires only a few hours. CONCLUSIONS: The EDB, together with the application of the ZSR model in data treatment, appears to be a valuable tool for studying the hygroscopic properties of pharmaceutical aerosols. PMID- 9327444 TI - Simultaneous quantification of an enantiomer and the racemic compound of ibuprofen by X-ray powder diffractometry. AB - PURPOSE: An X-ray powder diffractometric method was developed for the simultaneous quantification of the relative amounts of the racemic compound (+/-) of ibuprofen (I) and S(+)-ibuprofen (II), when they occur as a mixture. METHOD: The X-ray powder diffraction patterns of I and II show pronounced differences. This formed the basis for the determination of the relative amounts of I and II when they occur as a mixture. X-ray lines with d-spacings of 14.41 and 4.37 A were unique to I and II, respectively. Mixtures containing different proportions of I and II were prepared which also contained lithium fluoride (III) as an internal standard. RESULTS: A linear relationship was obtained when the intensity ratio (intensity of the 4.37 A line of II/intensity of the 2.01 A line of III) was plotted as a function of the weight fraction of II in the mixture. Similar results were obtained in the case of I. Using these standard curves, the weight fractions of I and II in "unknown" mixtures were determined. The experimentally determined analyte concentration ranged between 98 and 104% of the true value. The relative error in the analyses of individual samples was < 10%. The minimum detectable weight fraction of I and II and II in I were 0.032 (3.2% w/w) and 0.034 (3.4% w/w), respectively. The minimum quantifiable weight fractions were 0.136 for I and 0.112 for II. Since the X-ray diffraction patterns of S(+) ibuprofen and R(-)-ibuprofen are identical, the conclusions drawn regarding mixtures of I and II will also hold true in the quantitative analyses of mixtures of I and R(-)-ibuprofen. PMID- 9327445 TI - Effect of ionic strength on solution stability of PNU-67590A, a micellar prodrug of methylprednisolone. AB - PURPOSE: PNU-67590A is a water-soluble micellar prodrug of methylprednisolone (MP). The major products of degradation of PNU-67590A are MP by hydrolysis and methylprednisolone 17-suleptanate (17-E) by 21-->17 acyl migration. The effect of ionic strength on micelle formation and stability of PNU-67590A in aqueous solution was examined. METHODS: PNU-67590A solutions at pH 2 and 8 and ionic strength of 0.05, 0.1, 0.2, and 0.4 M were maintained at 25 degrees C in the dark to measure MP and 17-E levels over time. RESULTS: The rate of degradation of micellar PNU-67590A at pH 8 was less than that of monomeric PNU-67590A, and vice versa at pH 2. Increase in ionic strength decreased both the critical micelle concentration of PNU-67590A and the degradation of micelle PNU-67590A at both pHs, resulting in improved overall stability of PNU-67590A. CONCLUSIONS: Formulation of PNU-67590A in a concentrated solution with high ionic strength will maximize stability and shelf-life. PMID- 9327446 TI - ESR dosimetry of irradiated ascorbic acid. AB - PURPOSE: As an alternative to heat and gas exposure sterilization, ionizing radiation is gaining interest as a sterilization process for medicinal products. The aim of this work was to develop equations to describe the ESR curves versus dose and storage time after gamma irradiation of ascorbic acid. Several ESR data sets previously acquired in this laboratory were adopted to check the performance of the models. RESULTS: Limit of detection and limit of discrimination are respectively 0.5 kGy and 2 kGy for ascorbic acid. Linear regression is applicable for doses lower than 25 kGy. Since the radiation dose selected must always be based upon the bioburden of the products and the Degree of Sterility required (ANSI/AAMI/ISO 11137), doses in the range 5-25 kGy could be investigated and linear regression would appear to be the least expensive route to follow and gives good results. Quadratic fit, power function, exponential function and bi exponential functions are of more general applicability to predict irradiation dose. Decay kinetics for radicals versus storage were considered. Nonhomogeneous kinetics with time-dependent rate (diffusion-controlled second-order reaction) and bi-exponential function appeared valid to reproduce the experimental data. Discrimination between irradiated and unirradiated ascorbic acid is possible after a storage of 800 days. CONCLUSIONS: It is worth noting that, at present, ESR is the only technique which proves to be suitable for identification and quantification purposes in irradiated pharmaceuticals. Moreover, other features such as sensitivity, precision, ease and non-destructive readout make ESR superior to other proposed analytical techniques. PMID- 9327447 TI - Radiation effects on dopamine and norepinephrine. AB - PURPOSE: Radiation sterilization is becoming increasingly popular for the sterilization of many pharmaceutical products. We have investigated the gamma radiation induced effects on dopamine and norepinephrine by ESR spectroscopy. RESULTS AND DISCUSSION: Equations to describe the evolution of the ESR curves versus doses and time of storage are presented. Linear regression is, for dopamine hydrochloride, applicable for doses ranging from 10 to 25 kGy. Since the radiation dose selected must always be based upon the bioburden of the products and the degree of sterility required, doses in the range 10-25 kGy could be investigated and linear regression would appear to be the least expensive route to follow and gives good results. The comportment of noradrenaline bitartrate is more complex and the use of linear regression would appear more hazardous especially for low doses. For doses higher than 25 kGy, a more general equation is required. Power function using only 2 parameters could give good results but must be validated. Decay kinetics for radicals versus storage were considered. Non-homogeneous kinetics with time dependent rate constant and bi-exponential function appeared valid to reproduce the decay of radicals for, respectively, dopamine and norepinephrine. CONCLUSIONS: It is worth noting that, at present, ESR is the only technique which proved to be suitable for identification and quantification purposes in irradiated pharmaceuticals. Moreover, other features such as sensitivity, precision, ease and non-destructive readout make ESR superior to other proposed analytical techniques. PMID- 9327448 TI - N-trimethyl chitosan chloride as a potential absorption enhancer across mucosal surfaces: in vitro evaluation in intestinal epithelial cells (Caco-2). AB - PURPOSE: Previous studies have established that chitosan hydrochloride and glutamate are potent absorption enhancers for large hydrophilic compounds across mucosal surfaces. However, these compounds lack solubility at neutral pH values. A partially quaternized and well-soluble derivative of chitosan, N-trimethyl chitosan chloride, was synthesized and the effects of this polymer on the transepithelial electrical resistance and permeability of intestinal epithelial cells were investigated in vitro. METHODS: N-trimethyl chitosan chloride was synthesized by reductive methylation and characterized with NMR. The effect of this polymer (1.0-2.5% w/v) on the transepithelial electrical resistance of intestinal epithelial cells, using Caco-2 cell monolayers, was investigated. Permeation of the hydrophilic model compounds [14C]-mannitol (MW 182.2), FITC Dextran (MW 4400) and the peptide drug buserelin (MW 1299.5), in the presence of N-trimethyl chitosan chloride (1.5-2.5% w/v), was followed for 3 hours. The transport process of the fluorescent marker, FITC-Dextran 4400, across the cell monolayers was visualised with confocal laser scanning microscopy. Viability of the cells was checked with the trypan blue exclusion technique. RESULTS: N trimethyl chitosan chloride was found to be a perfectly water-soluble, partially quaternized (about 12%) derivative of chitosan. This polymer (1.5-2.5% w/v) caused a pronounced and immediate reduction (25-85%) in the transepithelial electrical resistance of Caco-2 cells. Large increases in the transport rate of [14C]-mannitol (32-60 fold), FITC-Dextran 4400 (167-373 fold) and buserelin (28 73 fold) were demonstrated. Confocal laser scanning microscopy confirmed that N trimethyl chitosan chloride opens the tight junctions of intestinal epithelial cells to allow increased transport of hydrophilic compounds through the paracellular transport pathway. No deleterious effects to the cells could be demonstrated with trypan blue. CONCLUSIONS: The potential use of N-trimethyl chitosan chloride as an absorption enhancer across mucosal surfaces could be an important contribution towards the development of effective delivery systems for hydrophilic drugs. PMID- 9327449 TI - Cellular retention of liposome-delivered anionic compounds modulated by a probenecid-sensitive anion transporter. AB - PURPOSE: Drug carriers such as liposomes may enhance the intracellular delivery of therapeutic agents for infectious or neoplastic diseases. However, the mechanisms affecting cellular retention of liposome contents are understood poorly. We tested the hypothesis that retention of anionic compounds may be modulated by a nonspecific probenecid-sensitive anion transport mechanism, and that liposome composition may determine the impact of such transporters on drug retention by cells. METHODS: The fluorescent anionic dye hydroxy-pyrene-[1,3,6] trisulfonate (HPTS) was transferred to the cytoplasm of cultured CV-1 or J774 cells by direct needle-microinjection or by ATP-induced permeabilization of the plasma membrane, respectively, to investigate whether the cells have anion transport mechanisms capable of extruding HPTS from the cytoplasm. Cellular retention of dye was monitored in the presence and absence of the anion transport inhibitors probenecid or sulfinpyrazone. Liposomes containing HPTS were co labeled with tetramethylrhodamine-labeled phosphatidylethanolamine (Rho-PE) as a marker of liposome membrane fate, and uptake was investigated using J774 cells. RESULTS: Needle-injected HPTS underwent both sequestration in early endocytic vesicles and rapid extrusion into the extracellular medium. Probenecid or sulfinpyrazone reduced the extrusion of HPTS. Thus HPTS is a substrate for a probenecid-sensitive anion transporter in J774 and CV1 cells. After delivery via fluid liposomes composed of phosphatidylglycerol:phosphatidylcholine:cholesterol (3:7:5 mole ratio) and co-labeled with Rho-PE, cell-associated HPTS declined more rapidly than did Rho-PE. Exposure of cells to 5 mM probenecid doubled the quantity of HPTS retained by cells, without changing the retention of the Rho-PE membrane marker. In contrast, the effect of probenecid was negligible when gel phase liposomes of distearoylphosphatidylglycerol:cholesterol (10:5 mole ratio) were used. CONCLUSIONS: Probenecid-sensitive nonspecific anion transporters can mediate the extrusion of model anions delivered via liposomes. However, liposome composition modulates the amount of material subject to extrusion from cells, possibly by altering the endocytic compartment in which liposomes release their contents. PMID- 9327450 TI - Estimation of the relative contribution of the transcellular and paracellular pathway to the transport of passively absorbed drugs in the Caco-2 cell culture model. AB - PURPOSE: The objective of this investigation was to determine, using the Caco-2 cell culture model, the extent to which the paracellular and transcellular routes contributed to the transport of passively absorbed drugs. An effort was also made to determine the controlling factors in this process. METHODS: We selected a heterologous series of drugs with varying physicochemical parameters for the investigation. Effective permeability coefficients of the model drugs (naproxen, phenytoin, salicylic acid, chlorothiazide, furosemide, propranolol, diltiazem, ephedrine, and cimetidine), at pH 7.2 and pH 5.4, were estimated using confluent monolayers of Caco-2 cells. The biophysical model approach, based on molecular size restricted diffusion within an electrostatic field of force, used by Adson et al. (1,2), was employed to estimate the permeability coefficients of the ionized and unionized forms of the drugs for the paracellular and transcellular route. RESULTS AND CONCLUSIONS: The permeability coefficients of the acidic drugs was greater at pH 5.4, whereas that of the basic drugs was greater at pH 7.2 and the transcellular pathway was the favored pathway for most drugs, probably due to its larger accessible surface area. The paracellular permeability of the drugs was size and charge dependent. The permeability of the drugs through the tight junctions decreased with increasing molecular size. Further, the pathway also appeared to be cation-selective, with the positively charged cations of weak bases permeating the aqueous pores of the paracellular pathway at a faster rate than the negatively charged anions of weak acids. Thus, the extent to which the paracellular and transcellular routes are utilized in drug transport is influenced by the fraction of ionized and unionized species (which in turn depends upon the pKa of the drug and the pH of the solution), the intrinsic partition coefficient of the drug, the size of the molecule and its charge. PMID- 9327451 TI - The absence of accessible vitronectin receptors in differentiated tissue hinders adenoviral-mediated gene transfer to the intestinal epithelium in vitro. AB - PURPOSE: Adenoviral (Ad) vectors have been used as efficient tools for gene therapy in various tissues, whereas in some differentiated epithelium transduction efficiency is almost abolished. METHODS: Caco-2 cell monolayers were chosen as an in vitro model for the differentiated intestinal epithelium. Fluorescence-labeled adenoviral particles were used for binding studies to cell surfaces. Internalization receptors for adenoviral uptake were detected by a fluorescence-labeled vitronectin antibody. Gene expression was studied by using the beta-galactosidase reporter gene. All experiments were done on undifferentiated and differentiated Caco-2 cells. Furthermore, adenoviral particles were allowed to bind to differentiated Caco-2 monolayers followed by a trypsinization step that disintegrates the monolayers and result in a cell suspension. Gene expression was tested after reseeding the cells into dishes. RESULTS: The results from adenoviral binding studies, vitronectin immunofluorescence detection and gene expression are in good agreement and indicate that virion binding as well as the expression of internalization receptors almost disappear in fully differentiated cells. Nonetheless, adenoviral binding to differentiated monolayers seems to be sufficient to cause up to 53% gene expression, but only if internalization of the vector can be induced by disintegrating the monolayers and releasing free vitronectin receptors. CONCLUSIONS: These findings indicate that gene transfer to the intestinal epithelium utilizing adenoviral vectors is poor and ineffective, because of the lack of sufficient internalization receptors. If these receptors can be exposed in differentiated epithelium, transduction can be made more efficient. Alternatively, a viral vector must be developed whose uptake mechanism is independent of integrin receptor expression like the enteral virus Ad40, or Ad5 could be conjugated to ligands that trigger viral internalization by receptor mediated endocytosis. PMID- 9327452 TI - Enhanced anti-inflammatory effects of Cu, Zn-superoxide dismutase delivered by genetically modified skin fibroblasts in vitro and in vivo. AB - PURPOSE: The purpose of this work was to evaluate the anti-inflammatory effects of secretable human Cu, Zn-superoxide dismutase (hSOD) delivered by genetically modified skin fibroblasts in vitro and in vivo. METHODS: Rat skin fibroblasts were transfected with pRc/CMV-ILSOD including secretable SOD-coding cDNA. The effects of host and transformants on oxidative stress in vitro models using the xanthine/xanthine oxidase (X/XO) system were examined to study the paracrine SOD action. The anti-inflammatory effects by transplantation of host and transformants were evaluated in an acute inflammation model, carrageenin-induced paw edema, in rats. RESULTS: The transformants (ILSOD cells) secreted SOD protein into the extracellular space, and the extracellular SOD activity in ILSOD cells cultures was significantly increased compared with that in host cell cultures. ILSOD cells diminished the cytotoxic activity by X/XO in a paracrine fashion. These protective effects of ILSOD cells against X/XO-induced cytotoxicity correlated well with the decrease in lipid peroxidation in the damaged cells. The in vivo study showed that transplantation of ILSOD cell suspensions into the hind paw in rats inhibited carrageenin-induced paw edema for at least 7 days, and the degree and the durability of these inhibitory effects were dependent on the number of ILSOD cells transplanted. These inhibitory effects of ILSOD cell suspensions were reduced by co-administration of antiserum for hSOD. Furthermore, the healing of paw edema caused by carrageenin was markedly enhanced by transplantation of ILSOD cells into the edemics hind paw. CONCLUSIONS: The findings suggested that genetically modified skin fibroblasts are a suitable delivery system for obtaining an efficient and continuous supply of SOD to the target site, and this strategy may be a useful drug delivery system for therapeutic proteins. PMID- 9327453 TI - Kinetic analysis of transcytosis of epidermal growth factor in Madin-Darby canine kidney epithelial cells. AB - PURPOSE: The aim of this study is to clarify the intracellular fate and a rate limiting step in transcytosis of epidermal growth factor (EGF) in Madin-Darby Canine Kidney (MDCK) epithelial cells. METHODS: The kinetics of transcytosis of 125I-EGF was investigated. To examine the fate of EGF molecules bound to its receptor on the cell surface. 125I-EGF was prebound to the basal surface at 4 degrees C, followed by extensive washing and subsequent incubation at 37 degrees C in EGF-free medium. RESULTS: Saturable transport of 125I-EGF through the cell monolayer could only be observed from the basal to apical side. Most (approximately 90%) of the EGF molecules bound to the surface receptor are internalized with a half-life of 1-3 min, followed by intracellular degradation with a half-life of 20-50 min. The exocytosis of internalized EGF into the apical medium is much slower with a half-life of 130-250 min. Even when 125I-EGF was incubated with MDCK cells at 37 degrees C and washed with acid to remove cell surface 125I-EGF, intact 125I-EGF appeared in the basal medium with a half life of 160-170 min. CONCLUSIONS: The exocytosis of internalized EGF into the apical medium is a rate limiting step in EGF transcytosis. At least a small amount of internalized EGF is recycled. PMID- 9327454 TI - Kinetic analysis of tetraethylammonium transport in the kidney epithelial cell line, LLC-PK1. AB - PURPOSE: The aims of this study were to establish a kinetic means of analyzing the membrane transport of organic cations in renal epithelial cells, and to simultaneously evaluate drug interactions in apical and basolateral membranes. METHODS: Tetraethylammonium (TEA) transport was measured using LLC-PK1 cell monolayers grown on microporous membrane filters. After incubating the cells with unlabeled TEA or other drugs, apical or basolateral medium was changed to that containing labeled TEA, and transcellular transport and cellular accumulation were measured. Clearance from apical medium to cells (CL12), cells to apical medium (CL21), cells to basolateral medium (CL23) and basolateral medium to cells (CL32) were calculated based on a three compartment model. RESULTS: TEA was accumulated progressively in the monolayers from the basolateral side and was transported unidirectionally to the apical side. CL32 was greater than CL12 and CL23 was greater than CL21. Therefore, the rate limiting step of TEA transport from the basolateral to the apical medium was the cell-to-apical step. Co incubation of TEA with procainamide decreased the transport parameters of TEA, CL12, CL21 and CL32, whereas that with levofloxacin decreased only CL12 and CL21, not affecting the parameters in basolateral membranes. CONCLUSIONS: Using a simple model, we analyzed the transport of organic cation in kidney epithelial cell line, LLC-PK1. This method can be useful for the analysis of cation transport and drug interactions in the apical and basolateral membranes of renal tubules. PMID- 9327455 TI - Perfusion cells for studying regional variation in oral-mucosal permeability in humans. I: Kinetic aspects in oral-mucosal absorption of alkylparabens. AB - PURPOSE: To evaluate regional differences in permeability of human oral mucosa. METHODS: Newly designed perfusion cells were used for the investigation. The cells were applied to 5 different sites, i.e., dorsum of tongue, ventral surface of tongue, labial mucosa, floor of mouth and buccal mucosa of human volunteers. Model drugs used were methyl-, ethyl-, propyl- and butylparaben, which are passively absorbed from oral mucosa and have different lipophilicities. Biexponential disappearance profiles of the alkylparabens were analyzed kinetically using a two-compartment linear open model. RESULTS: Both the partitioning parameters to the oral mucosa and the absorption rate constants to the blood circulation correlated to the lipophilicities of the compounds in all mucosa. As to the former parameter, no significant difference was recognized in all mucosa. While, the latter parameter exhibited the regional difference; the absorption rate constants in buccal mucosa were approximately one-half of those estimated in other oral mucosa. A positive relation was recognized between the retention in oral-mucosal compartment and the drug lipophilicity. CONCLUSIONS: The newly designed perfusion cells used in this study were useful to examine the regional variations of drug absorption from oral mucosa in humans. The absorption rate constant, the partition to oral mucosa and the residence time in oral mucosa increased with lipophilicity of the compound. The regional difference in the drug absorption process was demonstrated; the slow absorption and the prolonged retention were demonstrated in buccal mucosa. PMID- 9327456 TI - Polar solute transport across the pigmented rabbit conjunctiva: size dependence and the influence of 8-bromo cyclic adenosine monophosphate. AB - PURPOSE: To characterize the conjunctival permeability to polar solutes ranging from 182 to 167,000 daltons in molecular weight (m.w.). METHODS: Solute transport across the excised pigmented rabbit conjunctiva with a baseline transepithelial electrical resistance (TEER) of 1,285 +/- 46 ohm.cm2 was evaluated in the modified Ussing chamber under open-circuit conditions. The model solutes were mannitol (m.w. 182), 6-carboxyfluorescein (m.w. 376), and fluorescein isothiocyanate-labeled dextrans (FD4, m.w. 4,400-FD150, m.w. 167,000). RESULTS: For a given solute, the apparent permeability coefficient (Papp) was independent of solute concentration and direction of transport. As expected, the Papp decreased with solute size, from 27.7 x 10(-8) cm/sec for mannitol to 0.31 x 10( 8) cm/sec for FD150. When the experimental temperature was lowered from 37 degrees C to 4 degrees C. Papp decreased by approximately 50% for FD4 through FD40 and by > 80% for both FD70 and FD150. Equivalent pore analysis, assuming restricted solute diffusion via cylindrical, water-filled pores across the isolated tissue, revealed a radius of 5.5 nm at a pore density of 1.9 x 10(8) pores per cm2. The addition of 1 mM 8-bromo cyclic adenosine monophosphate (8 BrcAMP), known to stimulate Cl- secretion and decrease TEER, to the mucosal side of the conjunctiva increased the transport of mannitol, FD4, and FD40 by 28%, while not affecting FD150 transport. CONCLUSIONS: Our findings suggest that polar solutes up to FD40 traverse the conjunctival epithelial barrier primarily by restricted diffusion through equivalent pores of 5.5 nm radius and that solute movement is affected by reduction of TEER. On the other hand, polar solutes of the FD70 or larger may cross the barrier primarily via non-diffusional pathways such as non-specific endocytosis. PMID- 9327457 TI - The effect of current on skin barrier function in vivo: recovery kinetics post iontophoresis. AB - PURPOSE: The objective of this study was to determine the extent to which current passage perturbed the skin's intrinsic permeability, and to quantify how quickly and to what extent the barrier properties recovered from the effects of iontophoresis. METHODS: Laser scanning confocal microscopy (LSCM) and impedance spectroscopy (IS) were employed, respectively, to visualize and quantify the recovery kinetics. RESULTS: LSCM images were obtained following passive calcein diffusion through pre-iontophoresed HMS skin in vivo that had been allowed to recover for progressively longer periods of time. IS was used to quantify the rate and extent of skin permeability recovery following current pretreatment. Impedance spectra were recorded 0, 3, 5, 7, 9 and 18 hrs after current termination. CONCLUSIONS: Enhanced calcein permeability as assessed by confocal microscopy persisted for up to 24 hrs following current passage. Consistent with these LSCM findings, IS indicated that the time required for the impedance of hairless mouse skin to return to pre-iontophoresis levels (following 2-hr current passage at 0.5 mA/cm2) was at least 18 hrs. PMID- 9327458 TI - Iontophoretic delivery across the skin: electroosmosis and its modulation by drug substances. AB - PURPOSE: The long-term objective of this research is to understand how the efficiency of iontophoresis depends upon the structural and physicochemical properties of the administered drug. Specifically, the ability of certain drug species to alter the permselective properties of the skin was examined. METHODS: Using conventional in vitro methodology, the inhibition of electroosmotic flow induced by the iontophoresis of five different beta-blockers (of varying lipophilicity) was examined. The concomitant electrotransport of the most lipophilic species (propranolol) and the convective movement of solvent in the anode-to-cathode direction were measured. In addition, the possibility that electroosmosis might be augmented by the delivery of anionic drugs was also considered. RESULTS: Iontophoresis of lipophilic, cationic beta-blockers caused a concentration-dependent inhibition of conventional electroosmosis. The most hydrophilic analogs elicited no effect. As a result of this charge neutralization phenomenon, the optimal concentration for propranolol iontophoresis was significantly less than the maximum achievable in aqueous solution. Only a very modest improvement in convective solvent flow was induced by the cathodal iontophoresis of anionic compounds. CONCLUSIONS: The permselectivity of the skin can be altered by drugs which are positively charged and which possess a significant, adjacent hydrophobic surface. The latter seems able to "anchor" the molecule in the skin and the counter charge to the membrane's negative character ensures a tight association. Both lipophilicity and a positive charge are essential-without either, the phenomenon is not observed. The conformational flexibility of the drugs studied to-date, however, prevents unambiguous conclusions about the three-dimensional nature of the putative "binding site". PMID- 9327459 TI - Biphasic testosterone delivery profile observed with two different transdermal formulations. AB - PURPOSE: Our long-term goal is to develop formulations for pulsatile testosterone (T) delivery. T has been reported earlier to show biphasic pharmacokinetics in humans by Mazer et al, as well as biphasic permeation across excised rat skin by our group. We examined two kinds of formulations to evaluate their delivery profiles and to assess whether differences in the formulation approach affect pharmacokinetics in animal models. METHODS: One formulation consisted of T and a polymer blend dissolved in isopropanol; administered by dispensing the solution on the skin to cast a film in situ. The other was an adhesive-dispersion patch. In vitro release from the patch was evaluated using a flow-through cell interfaced with an HPLC pump and UV detector. Single dose pharmacokinetics were evaluated in castrated Wistar rats and bonnet monkeys immunized against gonadotropin-releasing hormone to deplete endogenous T. RESULTS: Two maximas were observed in the T release profile from the patch and in serum concentration versus time profiles in both animal models on application of either formulation. The relative magnitudes of the two maximas and the time interval separating them were different in the case of each formulation. CONCLUSIONS: Both formulations result in biphasic pharmacokinetics of T in the animal models studied. Discrete maximas presumably correlate with "burst" and "sustained" phases of drug release. PMID- 9327460 TI - Separation and quantitation of cationic liposome components by high performance liquid chromatography with evaporative light-scattering detection. PMID- 9327461 TI - Particle size distribution of a suspension aerosol using Andersen and Marple Miller cascade impactors. PMID- 9327462 TI - X-ray scattering analysis of human stratum corneum treated by high voltage pulses. PMID- 9327463 TI - Is the jejunal permeability in rats age-dependent? PMID- 9327464 TI - Evaluation of drug absorption after intrapulmonary administration using Xenopus pulmonary membranes: correlation with in vivo pulmonary absorption studies in rats. PMID- 9327465 TI - Review--animal waste used as livestock feed: dangers to human health. AB - Foodborne illness remains a common and serious problem, despite efforts to improve slaughterhouse inspection and food preparation practices. A potential contributor to this problem that has heretofore escaped serious public health scrutiny is the feeding of animal excrement to livestock, a common practice in some parts of the United States. In 1994, 18% of poultry producers in Arkansas collectively fed more than 1,000 tons of poultry litter to cattle, and the procedure is also common in some other geographic areas as a means of eliminating a portion of the 1.6 million tons of livestock wastes produced in the United States annually. While heat processing reliably kills bacterial pathogens, its use is limited by expense and other factors. Deep-stacking and ensiling are commonly used by farmers to process animal wastes, but the maximal temperatures achieved in stacked poultry litter are typically in the range of 43 to 60 degrees C (110 to 140 degrees F), below the inactivation temperatures of pathogenic salmonella and Escherichia coli species, and far below the USDA's recommended cooking temperatures of 71 to 77 degrees C (160 to 170 degrees F) for potentially manure-tainted meat products. In addition to the spread of potential pathogens, using animal wastes as feed presents the possibility that antibiotic-resistant bacteria may spread from one animal to another and that antibiotics or other chemicals may be passed between animals. Few research reports have addressed the safety of this practice, and those studies that have been published have generally been in controlled and artificial environments, rather than in on-farm conditions. Further microbiological studies are recommended to assess the extent of risk. PMID- 9327466 TI - The nocebo phenomenon: concept, evidence, and implications for public health. AB - The nocebo hypothesis proposes that expectations of sickness and the affective states associated with such expectations cause sickness in the expectant. The nocebo phenomenon is a little-recognized facet of culture that may be responsible for a substantial variety of pathology throughout the world. However, the extent of the phenomenon is not yet known, and evidence is piecemeal and ambiguous. This paper reviews the concept of nocebo and its association with the placebo phenomenon, gives examples of evidence for the nocebo phenomenon, and suggests public health implications. PMID- 9327467 TI - The nocebo effect: history and physiology. PMID- 9327468 TI - Nocebo: the power of suggestibility. AB - A useful way to summarize the placebo-nocebo theme is to consider the tension and interaction between conviction and responsibility. With the conviction of the mainstream biomedical paradigm prevalent today, it would be tempting to say to Dr. Engel's patient: "That question is nonsense. Cancer pain is not classified as 'male' or 'female.' Pain varies with location in the body and other factors." This response is technically honest but, in effect, it would have the impact of a nocebo. It would impair the patient's hope and morale. The doctor's honesty and conviction would serve as blinders to the patient's suffering. This type of honest statement results in a diminished sense of responsibility for the patient's well-being. Taking the biopsychosocial context into account, Dr. Engel achieved a balance between conviction and responsibility. The patient's question was understood within the meaning and metaphorical terms of her belief system. He answered in a manner that respected her private point of view toward pain and tapped her suggestibility, guiding her toward a probable placebo effect. "Female cancer" resonated with her personal beliefs and wish for less pain. Engel was both true to his convictions and responsible for providing the highest standard of care by understanding the patient's convictions and needs for comfort. The biopsychosocial concept provides a blueprint to bring the old-fashioned medical art of "humanness" to modern scientific care. Identifying the interactions of the problem, the person, and the totality of resources permits a focus on therapeutic strategies to promote placebo effects and prevent the consequences of nocebo. PMID- 9327469 TI - The importance of marital and socioeconomic status in incidence and survival of prostate cancer. An analysis of complete Norwegian birth cohorts. AB - BACKGROUND: Previous studies of the association between social and family status and prostate cancer (PCa) have given somewhat divergent results. Little attention has been paid to the possible importance of these factors for survival. METHODS: In this study, hazard regression models for PCa incidence and mortality were estimated on the basis of register- and census-based histories for complete Norwegian birth cohorts, giving a follow-up time of 16 million person years and 30,000 cases of PCa. RESULTS: A significant excess incidence of about 20% was found for ever-married men and for those with higher education. Marriage and socioeconomic resources appeared, however, to have a favorable effect on survival. Five-year relative survival from metastasized cancer among men with a high educational level was found to be 15 percentage points higher than among men with lower education. CONCLUSIONS: The observed differences in incidence are not easily explained. They apparently run counter to the hypothesis that multiple partners give a higher PCa risk, but may be consistent with the view that fat and meat consumption is risky. Better survival from PCa in higher socioeconomic groups and among married men may reflect differences in the search for, access to, or quality of treatment or a better constitution to fight the disease. PMID- 9327470 TI - Risk factors for low bone mineral density among females: the effect of lean body mass. AB - BACKGROUND: This study determines if body composition and lifestyle are risk factors for low radius-bone mineral density (R-BMD) and evaluates the role of body composition in the age-related decline of R-BMD. METHODS: Data on age, menopausal status, fat mass, lean body mass (LBM), drinking, smoking, and occupation were collected from 3,867 females ages 37-69, whose R-BMD was also measured. Multiple logistic regression analyses examined the predictive accuracy of these factors for low (20th percentile) R-BMD. RESULTS: Age, LBM, and menopausal status were risk factors for the 37-55 age group, while age, LBM, and lifestyle (alcohol consumption 3 or fewer days per week and currently smoking) were risk factors for the 56-69 age group. The odds ratio (OR) for LBM was nearly reciprocal to the ORs for age and for menopausal status. Our model has low sensitivity, high specificity, low positive predictive value, and high negative predictive value. CONCLUSIONS: Consumption of alcohol 3 or fewer days per week and being a current smoker have a negative effect on R-BMD among older (56-69) women. The positive effect of LBM on R-BMD continues from age 37 on LBM has an effect almost equal but opposite to those of aging and menopause on the risk of low R-BMD. High LBM after age 37 predicts normal R-BMD. PMID- 9327471 TI - The relation of smoking to waist-to-hip ratio and diabetes mellitus among elderly women. AB - BACKGROUND: Smoking is associated with lower body weight, but an increased risk of diabetes in some studies. Because smoking may increase waist-to-hip ratio (WHR), a risk factor for diabetes, we postulated that the relation between smoking and diabetes may be mediated in part by smoking-associated differences in body fat distribution. METHODS: We conducted a cross-sectional analysis of baseline data from 9,435 elderly nonblack women enrolled in the Study of Osteoporotic Fractures. Data were collected by Self-report and physical examination. Linear and logistic models were used to determine the relation of smoking to WHR and prevalence of self-reported diabetes. RESULTS: Current and past smokers had greater WHRs compared with never smokers. In multivariate models that adjusted for body mass index, the prevalence of diabetes was lower among smokers of < or = 10 cigarettes/day [odds ratio (OR) = 0.57, 95% confidence interval (CI) 0.31-1.03] and higher among smokers of > 10 cigarettes/day (OR = 1.38, 95% CI 0.99-1.92) compared with never smokers. The relation of smoking > 10 cigarettes/day to prevalence of diabetes was slightly attenuated after further adjustment for WHR. CONCLUSIONS: Smoking-associated differences in WHR may mediate, at least in part, the prevalence of diabetes among smokers of > 10 cigarettes/day. The decreased prevalence of diabetes that we observed among smokers of < or = 10 cigarettes/day was not explained by differences in obesity and requires confirmation. PMID- 9327472 TI - Factors influencing unawareness of hypertension among older Mexican Americans. AB - BACKGROUND: The objective of the study was to identify factors associated with unawareness of hypertension among Mexican Americans age 65 years and older. METHODS: This was a population-based survey of 3,050 older Mexican Americans conducted in five Southwestern states in 1993-1994. An in-home interview included sociodemographics, review of medications, and blood pressure measurements. RESULTS: Sixty percent of all subjects were hypertensive, and 37% of these were unaware of their diagnosis. Unaware hypertensive had significantly higher mean blood pressures than did aware hypertensives (145.7/ 86.2 mm Hg vs 142.4/83.1 mm Hg). While 77% of aware hypertensives were treated, only 10% of unaware hypertensives were treated. In multivariate analyses, factors associated with unawareness included male gender (OR = 1.8), being married (OR = 1.6), having Medicaid (OR = 1.6), having made fewer than two visits to a doctor in the past year (OR = 2.8), having a history of heart disease (OR = 0.57) or stroke (OR = 0.37), and having poor self-reported health (OR = 0.43). CONCLUSION: Despite 3 decades of hypertension detection and education programs, unawareness of hypertension remains high among older Mexican Americans. There is a continued need for community-based education programs for hypertensives who are unaware of their diagnosis, and also there is need for programs to increase access to primary care physicians. PMID- 9327473 TI - Prevalence and correlates of alcohol use among older adults. AB - BACKGROUND: The 1990 Health Promotion and Disease Prevention supplement to The National Health Interview Survey was used to develop point-prevalence data about drinking for four age groups, 55-64, 65-74, 75-84, and over 84, and to assess the impact of sociodemographics, health status, and health belief variables on light, moderate, and heavy alcohol consumption. The number of observations in the unweighted sample was 12,819, and the weighted sample contained 51,046,521 observations. METHODS: The chi 2 and Cohran-Mantel-Haenszel tests were used to investigate prevalence patterns, and odds ratios were generated from logistic regressions. RESULTS: Eighty percent of the sample had had at least 12 drinks during their lifetime, and 46% reported drinking during the survey year. The modal category for the number of days a respondent drank during the survey was 1 4 days, and the modal amount consumed on days that a person drank was 1-3 drinks. Age, gender, race, education, city size, labor force participation, geographic region, health status, having diabetes, and health beliefs about the adverse effects of excessive drinking and being overweight were associated with alcohol consumption, although their effects were different by drinking level. CONCLUSIONS: Analyses of health behaviors among older adults must recognize the diversity within this age group. Studies of drinking should differentiate between the amount consumed. Health beliefs need to be included in subsequent studies of health behaviors among older adults. PMID- 9327474 TI - Smoking behavior in a Swiss urban population: the role of gender and education. AB - BACKGROUND: Smoking has become more prevalent among U.S. men and women and European men in lower socioeducational groups. The relation between socioeducational status and smoking among European women has been studied less. METHODS: A survey assessing the smoking behavior and educational level of 943 women and 961 men ages 35 to 74 years from Geneva, Switzerland, was conducted. RESULTS: The prevalence of never smokers has declined among younger women but has remained stable among men. More men than women have ever smoked, but the difference has decreased among younger generations. Ever smoking was more prevalent among women with secondary (47.6%, age-adjusted OR 2.03, 95% CI 1.29 to 3.18) or tertiary (46.6%, age-adjusted OR 1.83, 95% CI 1.13 to 2.97) education relative to women with primary education (30.7%). Among males, ever smoking was slightly more prevalent among lower levels of education. There were moderate differences in quit ratio (ex-smokers/ever smokers) across educational levels among women (trend P = 0.08). In contrast, men with tertiary education stopped smoking more often (63.6%) than those with secondary (54.2%) or primary (47.6%) education (trend P = 0.008). For most women, primary education was associated with a later age at start of smoking while the inverse was true for men. CONCLUSION: Smoking behavior is evolving across generations of women in Geneva. It is more prevalent among educated women of the older generations, but this is less so among the younger generations. Women from Geneva may be currently experiencing the transition of smoking from upper to lower social classes. PMID- 9327475 TI - Integrating pros and cons for mammography and Pap testing: extending the construct of decisional balance to two behaviors. AB - BACKGROUND: The ability to study health-related behaviors in combination rather than singly will lead to a more comprehensive approach to health promotion. This investigation focused on mammography and Pap testing. One index was created to reflect the recency of receiving both examinations. A second index integrated opinions about the two procedures into a single measure, guided by the pros, cons, and decisional balance constructs of the Transtheoretical Model of behavior change. METHOD: Data were drawn from the baseline and 1-year follow-up surveys of an HMO sample of women ages 40-74 (N = 1,605). Data collection occurred by telephone. A series of analyses examined whether recency of screening was associated with opinions about screening. RESULTS: The first analysis showed that recency of Pap testing and whether or not a Pap test was obtained between the two surveys were associated with opinions about Pap testing. The next analysis examined the association between the indicator for regularity of both Pap testing and mammography, with the measure of opinions toward the two procedures. The variable measuring receipt of Pap test and mammography was associated with the combined measure of opinions. CONCLUSIONS: The ability to employ combined indicators for recency of testing and test-related opinions is promising for being able to take a more comprehensive approach to women's health. The paper discusses methodological considerations that arise when attempting to integrate two or more behaviors. PMID- 9327476 TI - Case-control studies in clinical research: mechanism and prevention of selection bias. AB - The mechanism by which selection bias occurs in case-control studies is explained to an audience of clinicians using a simple conceptual framework and a graphical presentation. A case-control study consists in comparing the frequency of exposure in a group of subjects having the studied disease (the cases) relative to another group free of that disease (the controls). Cases and controls can be thought of as arising from a hypothetical cohort study. Thus, enrolled cases are a fraction F1 of the exposed who developed the disease plus a fraction F3 of the unexposed who developed the disease during a given period. Similarly, enrolled controls are a fraction F2 of the exposed who did not develop disease plus a fraction F4 of the unexposed who did not develop the disease. A selection process is inherent to the design of case-control studies but it leads to selection bias only when the ratio of F1 x F4/F2 x F3 is not equal to unity. Examples demonstrate the implication of sampling fractions for designing and interpreting case-control studies performed in clinical settings. PMID- 9327477 TI - Validity of self-reported hypertension in the National Health and Nutrition Examination Survey III, 1988-1991. AB - BACKGROUND: The National Health and Nutrition Examination Survey (NHANES) is the main data source for hypertension surveillance. However, because of a gap of almost 10 years between each NHANES, self-reported data from annual surveys need to be examined as an alternative data source. This study analyzes the validity of self-reported hypertension in a national sample of non-Hispanic whites, non Hispanic blacks, and Mexican-Americans. METHODS: Sensitivity, specificity, and predictive values positive (PVP) and negative (PVN) of self-reported hypertension were calculated against two definitions of hypertension: the definition recommended by the Third Joint National Committee on Hypertension, JNC III (blood pressure > or = 140/90 and/or taking antihypertension medication) and a broader definition including control with lifestyle modifications. Data used come from the NHANES III, 1988-1991. RESULTS: Overall test characteristics using the JNC III definition are sensitivity 71%, specificity 90%, PVP 72%, and PVN 89%. Test characteristics were consistently higher for the broad than for the JNC III definition. Validity of self-reported hypertension is higher among women than among men and among persons with a medical visit during the past year than among those with no visits: validity was lowest among Mexican-American men. Due to the similarity between sensitivity and PVP, the prevalence of self-reported hypertension is nearly equal to the prevalence of JNC III-defined hypertension. CONCLUSIONS: Self-reported hypertension may be used for surveillance of hypertension trends, in the absence of measured blood pressure, among non Hispanic whites and non-Hispanic black women and persons with a medical visit in the past year. Validation should be repeated with each NHANES. PMID- 9327478 TI - Effect of response to a low-fat diet among adolescent males on their adult blood cholesterol levels. AB - BACKGROUND: While primary prevention of adult cardiovascular diseases should begin early, there are problems in identifying children at increased risk of future disease. METHODS: We did a follow-up study in 1991-1992 of 100 male former students at a boarding high school who had blood cholesterol measured in 1970 1971 both prior to and following a school-wide, reduced-fat dietary intervention. We compared adult cholesterol levels of the 50 subjects whose cholesterol decreased > or = 16.5% (the median decrease) following the 1970-1971 intervention (Diet-Sensitive) with the 50 whose response was < 16.5% (Non-Diet-Sensitive). RESULTS: Blood cholesterol of adults who were Diet-Sensitive in 1970-1971 was 4.2 mg/dl lower than their baseline values in adolescence, while adults classified as Non-Diet-Sensitive as adolescents showed a 15.9 mg/ dl increase in cholesterol over 21 years. Adjusting for baseline adolescent values, Non-Diet-Sensitive subjects were 4.8 (95% CI 1.4, 15.9) times as likely as Diet-Sensitive subjects to have adult cholesterol > or = 200 mg/ dl. Also, Diet-Sensitive adults on a low fat diet had adult blood cholesterol levels > 20 mg/dl lower than Non-Diet Sensitive adults on a similar diet (180.1 vs 202.1 mg/dl, respectively). CONCLUSIONS: Degree of response to a low-fat, low-cholesterol diet during adolescence may identify male subjects who will have differing patterns of cholesterol change over time. PMID- 9327479 TI - Transitions out of high school: time of increased cancer risk? AB - BACKGROUND: The effectiveness of lifestyle behavior interventions with children to reduce chronic disease risks in adulthood assumes stability in the lifestyle behaviors across time. The transition out of high school is a time when many changes occur in social roles, e.g., changing schools, leaving the parents' home, changing peers, finding employment, getting married, and becoming a parent. Cancer risk behaviors may increase as a result of some of these social role changes. METHODS: Concepts relevant to the stability or change in lifestyle behaviors through the transition out of high school are presented. Literature concerning diet, smoking, smokeless tobacco, alcohol, physical activity, sexual practices, and sun exposure behaviors through the transition is reviewed. RESULTS: Most lifestyle behaviors display increasing cancer risk around the transition out of high school. Different levels of change were associated with different pathways through the transition. Inconsistent findings were obtained in the pattern of co-occurrence of these behaviors. CONCLUSION: Priority research includes establishing the pattern of co-occurrence of lifestyle behaviors through the transition, identifying the pattern of tracking of each behavior through the transition, and identifying the primary influences on the group values and tracking of the behaviors. Longitudinal research is needed to control for preexisting differences between pathways through the transition. PMID- 9327480 TI - Dietary differences with green tea intake among middle-aged Japanese men and women. AB - BACKGROUND: Although several epidemiologic investigations have suggested a protective role of green tea against cardiovascular diseases and cancer, few studies examined how consumption of green tea was associated with intake of other dietary factors. METHODS: In the winters of 1989-1991, 880 men ages 40-49 years were randomly sampled from the general populations of five Public Health Center districts of Japan. Response rate was 72% (n = 634). A convenience sample of 373 spouses also consented to participate. They were interviewed on the frequency of consumption of green tea and 37 food items. A 3-day weighed food record was collected from a subgroup of the subjects (207 men and 164 women) to calculate daily intake of 22 nutrient variables. Consumption of the foods and nutrients was compared with three levels of green tea intake (< 1, 1-4, and > 4 cups/days) after adjustment for potential confounders. RESULTS: Among men, green tea was associated significantly with consumption of 10 foods (P < 0.05) and at borderline significance with 4 nutrients (P < 0.1). These foods and nutrients included fruits (apple, orange juice), vegetables (green, yellow, and pickled), total lipid, cholesterol, and carotene. Among women, green tea was associated with 6 foods and total energy. CONCLUSION: The results indicate that consumption of green tea is associated with diets that could modify the risks of cardiovascular diseases and cancer, especially among men. When the health effects of green tea are examined by observational epidemiologic studies, potential confounding and effect modification by other dietary factors should be controlled thoroughly. PMID- 9327481 TI - Morbidity on Kauai before and after Hurricane Iniki. AB - BACKGROUND: On September 11, 1992, Hurricane Iniki, a Class III/IV storm, passed directly over Kauai. This study is the first attempt to measure increases in injuries and other health outcomes among an entire population in the impact zone of a hurricane. METHODS: Medical chart data were abstracted from all facilities providing primary and emergency care on Kauai. Incidence of injury, cardiovascular disease, and asthma for the 2-week period following Hurricane Iniki were compared to those for the 2-week period preceding Iniki. RESULTS: A total of 1,584 injuries were treated in the post-Iniki period compared with 231 injuries treated in the pre-Iniki period (relative risk = 6.86, 95% confidence interval 5.98-7.87). Open wounds constituted over half of these injuries. Physician visits for asthma and cardiovascular disease were also significantly increased in the post-Iniki period (relative risks, respectively: 2.81, 95% confidence interval 1.93-4.09; 2.73, 95% confidence interval 1.51-4.94). CONCLUSIONS: Significant increases in the incidence of injuries, asthma, and cardiovascular disease occurred following Hurricane Iniki. Although no changes occurred in the proportion of patients needing hospitalization, additional injuries and illnesses after a natural disaster can burden existing medical facilities in a rural community with limited resources. Disaster preparedness plans need to include methods to increase services and supplies at existing medical facilities. PMID- 9327482 TI - The relationship of conjoint traits of dyslipidemias between young offspring and their parents in a community-based sample. AB - BACKGROUND: The relationship of dyslipidemias between young offspring and their parents was examined to evaluate its usefulness in predicting lipid disorders among parents and children. METHODS: Young offspring ages 5-17 years and their parents were studied in a community-based sample of 477 families. The dyslipidemias were defined as: (1) isolated high low-density lipoprotein cholesterol (LDL-C); (2) isolated high triglycerides (TG) and/or low high-density lipoprotein cholesterol (HDL-C); and (3) combined, involving both above. RESULTS: Children of parents with a given dyslipidemia type had the highest frequency of the same disorder (P < 0.001 to P < 0.05). In discriminant analyses only the corresponding disorders in their parents were selected into the models as significant predictors after controlling parental obesity. In terms of sensitivity, 54.8, 50.0, 66.7, and 69.1% of offspring could be correctly predicted for isolated TG/HDL-C, isolated LDL-C, combined, and any type of disorder, respectively, by the corresponding disorders in both parents. Likewise, the predictability of parent's dyslipidemia from their children's disorder was also modest. CONCLUSION: The conjoint dyslipidemias have familial basis to provide rationale for parents or children to determine their own risk status; however, sensitivity and positive predictive values are not high enough to be useful as a selective screening tool. PMID- 9327483 TI - Clustering of health-related behaviors among 18-year-old Australians. AB - BACKGROUND: Few studies among young adults have examined clustering of health behaviors affecting risk for lifestyle diseases. METHODS: Smoking, alcohol consumption, physical activity, and diet were examined among Australian 18-year olds (301 males, 282 females) initially recruited at the age of 9 years from 26 schools. Association analysis was used to recognize behavior clustering. RESULTS: Fat intake was greater among male smokers than nonsmokers (36% energy vs 34% energy). Women smokers ate less fiber (14.1 g/day) than did nonsmokers (17.8 g/day). Smoking was significantly related, among males, to unsafe drinking (odds ratio 2.38) and higher fat intake (odds ratio 1.06) and, among females, to unsafe drinking (odds ratio 1.59), lower dietary fiber (odds ratio 0.93), and less physical activity (odds ratio 0.36). Cluster analysis defined separate behavior clusters for men and women with smoking status identifying further subgroups. Smoking, drinking alcohol to excess, and adverse dietary choices clustered among men and women, with physical inactivity also clustering among women. CONCLUSION: Smoking among adolescents is an important indicator of behaviors influencing risk for later cardiovascular disease and other medical disorders. Multimodal approaches allowing for gender differences in health-related behaviors are likely to be more successful than targeting a single behavior in this age group. PMID- 9327484 TI - Determinants of obesity in relation to socioeconomic status among middle-aged Swedish women. AB - BACKGROUND: It has been previously demonstrated that obesity is common among women with low socioeconomic status (SES), but the factors accounting for this association are not well known. According to our hypothesis, low SES is associated with psychosocial stress, an unhealthy lifestyle, and reproductive history, which may increase the likelihood of women with low SES to be overweight or obese. METHODS: We examined overweight and obesity in relation to SES among 300 healthy women ages 30-65 years, who constitute the control group of the Stockholm Female Coronary Risk Study, a population-based case-control study of women with coronary heart disease. This control group was compared with a large population-based sample and found to be representative of healthy Swedish women ages 30-65 years. We used an aggregate of education and occupation as a measure of SES and defined overweight as body mass index (BMI) between 23.8 and 28.6 kg/m2 and obesity as BMI > 28.6 kg/m2. RESULTS: Low SES was a strong determinant of overweight and obesity among middle-aged healthy Swedish women. The odds of being overweight or obese increased with lower social position. After adjustment for age, the odds ratios for overweight and obesity among women in a low vs high position were 2.2 [95% confidence interval (CI) 1.1 to 4.4) and 2.7 (95% CI 1.1 to 6.7), respectively. Both low social position and obesity were related to reproductive history (higher parity and earlier age at menarche), unhealthy dietary habits, and unfavorable psychosocial factors (poor quality of life, low self-esteem, and job strain). These factors together explained 53% of the low-SES obesity association. CONCLUSIONS: Reproductive history, unhealthy dietary habits, and psychosocial stress accounted for a large part of the association between low SES and obesity. Dietary habits and psychosocial stress are potentially modifiable factors, which should be taken into account in intervention programs among women with low SES. PMID- 9327485 TI - Evaluation of two strategies for heart health promotion by direct mail in a low income urban community. AB - BACKGROUND: The objective of this study was to evaluate the reach of mass mailings of heart health education print materials in a low-income, urban community. Materials included a monthly newsletter and a self-help behavior change kit, both distributed to all 12,789 households in the study community. METHODS: Recall, use, and self-reported impact of the materials were measured in a cross-sectional survey of a random sample of 345 adults conducted 2 weeks after distribution of the kit and 18 months after delivery of the first newsletter. RESULTS: Over one-third of the subjects (38.6%) recalled the newsletter and 27.9% had read one or more newsletters; 21.7% recalled the kit and 10.8% had read it. Few subjects had read both materials. Female gender and older age were independent correlates of having seen and read the newsletters. Older age, being widowed/separated/divorced, and infrequent physical activity were correlates of having seen and read the kit. CONCLUSIONS: Although the newsletter and kit formats might appeal to different segments of the population, mass mailings of heart health education print materials in a low-income urban community can reach large numbers of individuals. The cost effectiveness of repeated mailings of short, simple newsletters might be higher than a single mailing of a more complex behavior change kit. PMID- 9327486 TI - Cultural, material, and psychosocial correlates of the socioeconomic gradient in smoking behavior among adults. AB - BACKGROUND: The aim was to identify the correlates of educational differences in smoking among adults. METHODS: We used data from the baseline of a Dutch longitudinal study, relating to a population of 2,462 respondents, ages 25-74. Logistic regression was used to assess the educational gradient in smoking. Current smokers were compared with former and never smokers, respectively. RESULTS: The risk of being a current smoker compared with being a former/never smoker was higher among lower educational groups. For example, the odds of being a current smoker compared with never smoker among persons in the lowest level was more than five times as high as that for persons in the highest level. A substantial part (20-40%) of the increased risk of being a smoker among lower groups appeared to be associated with adverse material conditions. The financial situation especially accounted for that effect. One of the cultural factors, i.e., locus of control, was found to account for approximately 30% of the educational gradient in the case in which smokers were compared with former smokers. Psychosocial factors, i.e., neuroticism and coping styles, accounted for less of the gradient in smoking than cultural and material factors. CONCLUSIONS: On the basis of the results, we hypothesize that both cultural and material factors contribute to the higher smoking rates among lower socioeconomic groups. Psychosocial factors seem to be less important. If our results are confirmed in more powerful studies, this would indicate, first, that possibilities for a reduction of smoking differences may be found in tailoring smoking cessation programs to the more externally oriented locus of control and the coping styles that are common among lower educational groups, and second, that a reduction of smoking differences may follow from an improvement of the material living conditions of lower socioeconomic groups. PMID- 9327487 TI - Priorities in behavioral research in cancer prevention and control. PMID- 9327488 TI - Behavioral research contributions and needs in cancer prevention and control: adherence to cancer screening advice. AB - BACKGROUND: Research has been critical in understanding human behavior related to the early detection of cancer. METHODS: Based on the literature and the author's experience, this paper reviews behavioral research accomplishments in the past decade and needs for the future. RESULTS: Accomplishments have included an improved understanding of the barriers to screening, methods to improve the provision of tests by practitioners, and progress in the development and application of community interventions. Outstanding needs are summarized in 12 areas that include a continued focus on underserved populations and an examination of the use of screening tests in the presence of incomplete evidence of efficacy, translational research that explores the behavioral consequences of genetic susceptibility testing, continued development and use of computer-based technologies in the delivery of preventive services, and studies of the potential negative consequences of screening. Critical changes in the delivery of health care in the era of managed care call for increased behavioral research in health services and health care policy. CONCLUSION: Although this review of research needs is not exhaustive, it is clear that the challenges and opportunities for behavioral research are substantial, and this effort will be a critical part of the overall national research agenda for cancer control. PMID- 9327489 TI - Behavioral research to enhance adjustment and quality of life among adults with cancer. AB - The purpose of this article is to recommend future directions for behavioral research to enhance adjustment and quality of life for adults diagnosed with cancer. As context for the recommendations, the domain of behavioral research in psychosocial oncology is briefly described, the state of the science measuring quality of life is summarized, and research results from behavioral research on quality of life are reviewed. PMID- 9327490 TI - Prevention and control research within a changing health care system. AB - The agenda for behavioral and social science research in cancer prevention and control needs to be assessed and interpreted within the context of a changing health care system. These changes present potential opportunities and barriers to an evolving research agenda. Opportunities include access to defined populations and providers, emphasis on clinical outcomes, and access to clinical and financial data. Barriers include intense competition among providers and an overriding emphasis on cost containment. To meet these challenges, attention needs to be given to developing new partnerships with provider organizations, emphasizing interdisciplinary cooperation and training, and reassessing underlying behavioral assumptions and models. PMID- 9327491 TI - The behavior-biology interface in cancer prevention and control science. PMID- 9327492 TI - Behavioral research contributions and needs in cancer prevention and control: dietary change. AB - This paper summarizes key behavioral research contributions to the promotion of healthful diets and identifies the outstanding behavioral research needs that could lead to positive dietary changes in the United States. Nutrition plays an important role in the initiation, promotion, and progression of cancer. Dietary guidelines for health promotion and cancer prevention recommend diets that are lower in fat and higher in fiber, fruits, and vegetables. Behavioral research on dietary change has become more rigorous and sophisticated in the past decade, with noteworthy contributions in four areas: behavioral research within clinical trials, self-help or minimal contact intervention strategies, school nutrition programs and services, and advances in the development of measures. Work in progress includes large-scale randomized intervention trials, with the majority of funding for studies to increase fruit and vegetable consumption. There are many needs for further research. Six priority areas for behavioral research are identified and discussed: (1) determinants of dietary behavior and change processes; (2) policy, environmental, and organizational interventions; (3) studies of dietary change and exercise and interventions with persons at high risk for diet-related cancers; (4) methodological research; (5) research on diffusion and dissemination; and (6) systematic behavioral research on dietary change in clinical trials. A concerted research effort in the area of dietary change has great potential benefits for cancer prevention and control and for public health in general. PMID- 9327493 TI - Behavioral research contributions and needs in cancer prevention and control: tobacco use prevention and cessation. AB - Following a brief review of the etiology and prevalence of tobacco use and data on the effectiveness of prevention and cessation interventions, recommendations for a research agenda are outlined. It is suggested that research on youth tobacco initiation and cessation be given highest priority because of rising prevalence rates, fundamental social importance, and the widespread support for such efforts. Policy and community approaches to deterring youth tobacco use deserve particular attention. Adult intervention research should focus on health care settings and include factors that both help and hinder adoption and routine implementation of tobacco interventions by clinicians. Developing and evaluating practical ways of using nicotine replacement therapies or other pharmacological therapies in primary care are also of importance. Media interventions that segment the smoking population by age, ethnicity, and developmental milestones should be encouraged. Three approaches could profit from working conferences of investigators and other interested parties to review the data and suggest research directions: worksite interventions, interventions with ethnic populations, and matching or tailoring interventions to specified characteristics of smokers. The importance of devoting considerable resources to investigator initiated contrasted with sponsor-directed research is discussed. PMID- 9327494 TI - Translational behavioral research in cancer genetics. AB - Recent advances in the molecular genetics of cancer have created new opportunities for behavioral scientists interested in decision-making, risk communication, and health behavior. As part of a National Cancer Institute sponsored workshop, behavioral scientists participated in breakout groups charged with generating suggestions for research priorities. The following five research priorities were identified for the area of genetic testing for cancer susceptibility: (1) enhancing informed decision-making about whether to be tested, (2) improving methods of cancer risk communication, (3) examining the psychosocial, clinical, and economic outcomes of testing, (4) evaluating the efficacy of psychological and behavioral interventions, and (5) evaluating approaches to prevent cancer in mutation carriers. Across each of these areas, family issues, ethnic and cultural issues, and economic implications should be evaluated. PMID- 9327495 TI - Atomic force microscope imaging of DNA and DNA repair proteins: applications in radiobiological research. AB - By using the atomic force microscope (AFM), three-dimensional structures of biological specimens may be imaged at nanometer resolution. Furthermore, samples can be imaged in air or in fluid environments. The tapping mode of AFM operation for imaging has offered a significant advance in visualizing soft biological structures, such as DNA, proteins, and membranes. Here, we review the principles underlying the application of this instrument to radiation biological investigations. We focus on examples of proteins involved in the processes of repair of damaged DNA, including poly(ADP-ribose) polymerase, Ku protein, and DNA protein kinase. Novel observations on the character of DNA damage and repair have been addressed by direct visualization of DNA and protein-DNA interactions, such as the observation that the Ku protein is capable of physically joining DNA fragments in vitro. The AFM offers a powerful tool for investigating biologically important molecular interactions that are relevant to DNA damage and repair processes. PMID- 9327497 TI - Potential indicators of radiosensitivity in squamous cell carcinoma of the head and neck. AB - Recently, attention has focused on the potential of oncogenes, tumour suppressor genes, and assessment of cell proliferation as biological response indicators in human cancer. In this study, immunocytochemical analysis was used to evaluate the usefulness of Ki67, epidermal growth factor receptor (EGFr), and the protein products of c-Myc and Bcl-2 as indicators of radiosensitivity in primary cultures of head and neck tumours. Primary cultures established from tumours taken at surgery were divided into two groups; the control group remained untreated, and the treatment group received a single dose of 2 Gy. The cultures were incubated for 14 days, after which time they were fixed and examined immunocytochemically. The response to treatment of the cultures was measured as the percentage of growth inhibition (%GI) in the treated cultures relative to the untreated controls. Expression of Ki67 measured after a single dose of 2 Gy significantly differentiated the radioresistant and radiosensitive groups (P = 0.045); high percentages of Ki67+ cells correlated with radioresistance. A significant difference was found between the expression of EGFr in the resistant and sensitive groups, as measured in control cultures and after a dose of 2 Gy (P = 0.002 and P = 0.03, respectively); high levels of expression of EGFr correlated with radioresistance. The level of expression c-Myc+ cells, as measured in control cultures, significantly distinguished the radiosensitive group from the radioresistant group (P = 0.05). These results indicate a potential role for these proteins as indicators of radioresistance. PMID- 9327496 TI - S-phase fraction, 5-bromo-2'-deoxy-uridine labelling index, duration of S-phase, potential doubling time, and DNA index in benign and malignant brain tumors. AB - Seventy-one histologically malignant brain tumors, 52 histologically benign brain tumors, and 14 cerebral metastases were characterized according to DNA content and proliferative capacity. DNA ploidy, DNA index (DI), S-phase fraction (SPF), 5 bromo-2'-deoxy-uridine (BrdUrd) labelling index (LI), duration of S-phase (Ts), and potential doubling time (Tpot) were assessed by flow cytometry (FCM). In histologically benign tumors, a high percentage of DNA diploid tumors and a low proliferative capacity in DNA diploid tumors were found. Histologically malignant tumors and cerebral metastases were both found to be characterized by a low percentage of DNA diploid tumors and a high proliferative capacity in DNA diploid tumors. The proliferative capacity of DNA aneuploid benign tumors and that of DNA aneuploid malignant tumors, however, appeared not to differ significantly. The number of DNA aneuploid tumors was small. Duration of S-phase was short (range 3.9-4.7 hr) and appeared not to differ between the three groups. From this, the observed differences in Tpot values should be accredited mainly to differences in LI. High-grade as well as low-grade gliomas both appeared to be characterized by malignant (FCM) features, i.e., 1) a high percentage DNA aneuploidy, 2) a high mean DI (for DI > 1), and 3) a high proliferative capacity. PMID- 9327498 TI - Prostate volumes and organ movement defined by serial computerized tomographic scans during three-dimensional conformal radiotherapy. AB - The aim of this study was to assess changes in prostate volumes and organ movement during a course of external beam irradiation using serial computerized tomographic (CT) scans and three-dimensional treatment planning software. Ten consenting prostate cancer patients underwent repeat CT scans at biweekly intervals during the course of external beam irradiation. The spacing of 5 mm was used because this spacing mimics our clinical treatment approach. Prostate locations were determined by merging CT images using bony anatomy and comparing the differences in the prostate volumes, the edges (anterior, posterior, superior, inferior, and lateral) and centers of the prostate (EoP and CoP, respectively). Compared to the 10 initial treatment planning CT scans, the prostate volume determined by the repeat CT scans tended to be smaller (approximately 14%, P < 0.001). The prostate volumes determined by repeat CT scans tended to be stable with a mean volume of 86% (S.D. = 18%) of the initial CT. When assessed by changes in the EoP, superior movements appeared to be the most common source for concern for adequate coverage of the prostate, while inferior movement was not seen. When assessed by changes in CoP, movement of > or = 3 mm was noted in 47% of the studies in the superior direction, with the average displacement being approximately 2.0 mm. In this study, the prostate volume tended to be smaller 2 weeks after the start of radiotherapy. Moreover, the prostate volumes defined by the serial CT scans were less reproducible than expected. Superior displacement of the prostate is the most common and significant type of displacement, while inferior movement is least frequent when patients are simulated with their rectums empty. Because of the magnitude of daily setup errors, organ movement, and problems with reproducibility in target definition, additional field edge reductions do not appear to be warranted during the delivery of three-dimensional conformal radiotherapy. Efforts should be directed at improving our ability to reduce organ movement and accurately targeting the prostate. PMID- 9327499 TI - Computed tomography-magnetic resonance image fusion: a clinical evaluation of an innovative approach for improved tumor localization in primary central nervous system lesions. AB - We describe our initial experience with the AcQSim (Picker International, St. David, PA) computed tomography-magnetic resonance imaging (CT-MRI) fusion software in eight patients with intracranial lesions. MRI data are electronically integrated into the CT-based treatment planning system. Since MRI is superior to CT in identifying intracranial abnormalities, we evaluated the precision and feasibility of this new localization method. Patients initially underwent CT simulation from C2 to the most superior portion of the scalp. T2 and post contrast T1-weighted MRI of this area was then performed. Patient positioning was duplicated utilizing a head cup and bridge of nose to forehead angle measurements. First, a gross tumor volume (GTV) was identified utilizing the CT (CT/GTV). The CT and MRI scans were subsequently fused utilizing a point pair matching method and a second GTV (CT-MRI/GTV) was contoured with the aid of both studies. The fusion process was uncomplicated and completed in a timely manner. Volumetric analysis revealed the CT-MRI/GTV to be larger than the CT/GTV in all eight cases. The mean CT-MRI/GTV was 28.7 cm3 compared to 16.7 cm3 by CT alone. This translated into a 72% increase in the radiographic tumor volume by CT-MRI. A simulated dose-volume histogram in two patients revealed that marginal portions of the lesion, as identified by CT and MRI, were not included in the high dose treatment volume as contoured with the use of CT alone. Our initial experience with the fusion software demonstrated an improvement in tumor localization with this technique. Based on these patients the use of CT alone for treatment planning purposes in central nervous system (CNS) lesions is inadequate and would result in an unacceptable rate of marginal misses. The importation of MRI data into three-dimensional treatment planning is therefore crucial to accurate tumor localization. The fusion process simplifies and improves precision of this task. PMID- 9327501 TI - At issue: are traditional societies schizophrenogenic? AB - Schizophrenia is apparently less common in traditional than in nontraditional societies, and the course of illness in these cultural settings may also be more benign. Viral, political, economic, social labeling, and other explanations have been offered over the years for these differences. In contrast to those ideas that suggest the presence of a schizophrenogenic stress in urbanized, Westernized populations, I propose that traditional societies are actually schizophrenogenic compared with nontraditional societies. Assuming a multifactorial threshold model for the development of schizophrenia, traditional societies may be characterized by a lower threshold for developing schizophrenia in at-risk individuals. In the short term, this leads to a greater proportion of all clinical cases being of a less severe variety; in the long term, genes predisposing individuals to develop schizophrenia are exposed to the effects of negative selection, ultimately resulting in a relatively lower level of overt schizophrenia in these populations. The greater social demands placed on individual actors in traditional societies and the lack of variability in social network size may contribute to the (relatively) schizophrenogenic environment. PMID- 9327500 TI - Warping CT scans from nontreatment to treatment position. AB - This paper describes a cost-effective technique that optimally utilizes all available diagnostic studies for three-dimensional treatment planning. A simulator unit modified to produce cross-sectional images (simulator-CT unit) is used to create a reference data set with the patient in the treatment position. Registration software (qsh) brings other diagnostic studies into agreement with this reference data set. Two cases are presented as examples of the use of this technique. Registration of abdominal scans from the same patient demonstrates the warping of a nontreatment position study to the treatment position. The second case is based on paired data sets through the head, in which the diagnostic study was obtained by using a gantry tilt to follow the base of the skull and to avoid sections passing through the teeth. The registration software provides a method for combining diagnostic studies into a single "master" data set. The success of the transformation depends on the operator's ability to identify corresponding anatomic landmarks for different data sets and on the magnitude of the variation in the patient's position from one procedure to the next. Limitations in image quality and the number of cross-sections obtainable from a simulator-CT unit can be partially overcome by using the described technique. Thus, the information contained in nontreatment position diagnostic tests can be used accurately for treatment planning at limited cost. PMID- 9327503 TI - The neuroanatomies of schizophrenia. AB - Schizophrenia is a brain disease whose pathophysiology has escaped detection despite intensive investigation. The failure to delineate the neuroanatomy of schizophrenia is related in part to both the subtle nature of the neuropathological abnormalities and to the failure to address adequately the pathophysiological heterogeneity of schizophrenia. The symptoms of schizophrenia aggregate into relatively independent symptom complexes, which suggests that there may be a distinct neural substrate for each complex. If this is true, then the neuroanatomy of schizophrenia is better addressed as the separate neuroanatomies of symptom complexes. However, the use of symptom complexes to guide future neuroanatomical investigations raises crucial methodological issues, including the differentiation of primary versus secondary symptoms, trait versus state characteristics, and continuous versus categorical variables. Decisive hypothesis testing requires that these issues be addressed in study design. PMID- 9327504 TI - Anatomy of schizophrenia revisited. AB - The search for an anatomy of schizophrenia has engendered an enormous, almost indigestible mass of data. In no studies do all patients show the same deviations from control samples. No morphological or microscopic abnormality has been found that is either necessary or sufficient for the diagnosis. In contrast to epilepsy, in which a proliferation of excitatory pathways or inadequate inhibitory factors are paramount, schizophrenia may represent a genetically and age-determined elaboration of one or more inhibitory networks in response to specific physiological events (e.g., the increased neuronal activity in limbic and hypothalamic structures during the physiological events of puberty) or to brain injury or defect. Current diagnostic classifications, including the positive-negative categories, have not led to separation of the disorder into etiologically or pathologically similar subgroups. Analysis of morphological and other biological pathology by a different nosological principle, such as trajectory of the illness, and separate correlation of anatomical and other biological outliers with clinical and demographic factors may be more successful strategies than pooling and averaging results from a mixture of patients diagnosed with schizophrenia. PMID- 9327505 TI - Postmortem studies of the hippocampal formation in schizophrenia. AB - Many postmortem studies report differences between the hippocampal formations of patients with schizophrenia and those of controls. These differences include volume changes, cell density changes, periventricular gliosis, senile degenerative changes, and abnormalities of neuronal size, position, or orientation. However, the findings are almost never common to all schizophrenia patients within a series. Furthermore, some well-designed studies are negative, and different positive reports are mutually contradictory. Some of the inconsistencies are methodological. The normal variation, over small distances, in the cytoarchitecture of the temporal allocortex creates particular difficulties when this region is studied with a limited number of sections, especially if the sample size is small. Other inconsistencies are probably the result of case selection. We review the methods and findings of some of these studies, stressing the dangers of eliminating (rather than evaluating) cases with definite neuropathologic changes. We conclude that the existing postmortem studies of temporal lobe morphology provide little conclusive evidence for the neural substrate of schizophrenia. PMID- 9327506 TI - Neuropathology of schizophrenia: cortex, thalamus, basal ganglia, and neurotransmitter-specific projection systems. AB - This article reviews neuropathological studies in the search for an anatomical correlate of schizophrenia. Replication of many results has proven to be difficult. A consistent finding is the lack of significant gliosis in the neocortex. Intriguing findings that need further corroboration include decreased volume and cell number of the mediodorsal thalamic nucleus, cytoarchitectural alterations of the prefrontal cortex and upper layers of the anterior cingulate gyrus, and superior temporal gyrus abnormalities. Most neuropathological studies investigate regional brain volume and cell density. Highly variable shrinkage of brain tissue postmortem makes these estimates prone to bias and often not comparable across studies. So far, no strong clinicopathological correlations and no pathological criteria to diagnose schizophrenia have been established. PMID- 9327507 TI - The temporolimbic system theory of positive schizophrenic symptoms. AB - This article proposes that subtle structural and functional disturbance of limbic key structures in the medial temporal lobe-especially of the left hippocampal formation and parahippocampal gyrus-can explain the so-called positive symptoms of schizophrenia. After presenting pathophysiological considerations linking limbic dysfunction to schizophrenia, the article reviews evidence from structural, biochemical, and functional studies supporting the theory. Also discussed here are neurodevelopmental and laterality aspects, as well as predictions, questions, and future tasks derived from the theory. PMID- 9327508 TI - Functional and anatomical aspects of prefrontal pathology in schizophrenia. AB - Clinical and experimental research have provided anatomical, pharmacological, and behavioral evidence for a prominent prefrontal dysfunction in schizophrenia. Negative symptoms and behavioral disorganization in the disorder can be understood as a failure in the working memory functions of the prefrontal cortex by which information is updated on a moment-to-moment basis or retrieved from long-term stores, held in mind, and used to guide behavior by ideas, concepts, and stored knowledge. This article recounts efforts to dissect the cellular and circuit basis of working memory with the goal of extending the insights gained from the study of normal brain organization in animal models to an understanding of the clinical disorder; it includes recent neuropathological findings that indicate that neural dystrophy rather than cell loss predominates in schizophrenia. Evidence from a variety of studies is accumulating to indicate that dopamine has a major role in regulating the excitability of the cortical neurons upon which the working memory function of the prefrontal cortex depends. Interactions between monoamines and a compromised cortical circuitry may hold the key to the salience of frontal lobe symptoms in schizophrenia, in spite of widespread pathological changes. We outline several direct and indirect intercellular mechanisms for modulating working memory function in the prefrontal cortex based on the localization of dopamine receptors on the distal dendrites and spines of glutamatergic pyramidal cells and on gamma-aminobutyric acid (GABA) ergic interneurons in the prefrontal cortex. Understanding the interactions between the major cellular constituents of cortical circuits-pyramidal and nonpyramidal cells-is a necessary step in unraveling the receptor mechanisms, which could lead to an effective pharmacological treatment of negative and cognitive symptoms, as well as improved insight into the pathophysiological basis of the disorder. PMID- 9327509 TI - The basal ganglia and cognitive pattern generators. AB - This article introduces the notion of cognitive pattern generators and suggests, by analogy with the central pattern generators of the motor system, that these pattern generators operate to organize neural activity underlying aspects of action-oriented cognition. It is further proposed that the basal ganglia are involved in the control of cognitive as well as motor pattern generators. Disorders of the basal ganglia may thereby contribute to neural circuit dysfunctions that are expressed as positive and negative symptoms of schizophrenia. PMID- 9327510 TI - The interface between dopamine neurons and the amygdala: implications for schizophrenia. AB - A substantial amount of research has focused on the midbrain dopamine system and its role in emotional and motivational behaviors. In diseases in which dopamine function is compromised, patients exhibit a constellation of symptoms, suggesting that the dopamine system plays an important role in the integration of several functions. Subgroups of dopamine neurons receive information from limbic and association areas and project widely throughout cortex and striatum, including motor areas. A dorsal tier of dopamine neurons receive input from the ventral (limbic-related) striatum and from the amygdala and project widely throughout cortex. A more ventrally located group of dopamine cells receives input from both the limbic and association areas of striatum and projects widely throughout the striatum, including the sensorimotor regions. Through these projections, the limbic system has an enormous influence on dopamine output and can therefore affect the emotional and motivational "coloring" of a wide range of behaviors. PMID- 9327511 TI - Cortical development and thalamic pathology in schizophrenia. AB - In this article, morphological data suggesting that brain development may be altered in schizophrenia are reviewed in relation to the major events in neural development. In the absence of severe defects in brain structure in individuals with schizophrenia, developmental processes governing the establishment, refinement, and maintenance of connections are potential sites of pathological involvement. Alterations in connectional patterns are likely to result in activity-dependent changes in gene expression for molecules involved in the neurotransmission process, with functional consequences. Loss of cells in the thalamus may be primary or secondary to cortical or other subcortical pathology. Loss of thalamic cells and/or of corticothalamic inputs could lead to disintegration of thought processes by a failure in functional brain states dependent on collective oscillation of large ensembles of cortical and thalamic neurons. PMID- 9327512 TI - What an archaeological dig can tell us about macro- and microcircuitry in brains of schizophrenia subjects. AB - This commentary on recent postmortem investigations suggests that schizophrenia may involve alterations of corticothalamic and temporolimbic regions of the brain. Although studies of this type are beginning to provide unique insights into the underlying pathophysiology of this disorder, all such investigations are generally hampered by the inability to differentiate between primary and secondary changes within complex macro- and microcircuitry. To overcome this basic epistomological problem, it will be necessary to develop novel strategies for determining how the communication between and within these various brain regions is decompensating, and later, compensating at different stages of the life cycle in schizophrenia. PMID- 9327513 TI - Neuroanatomy of schizophrenia. AB - The articles that appear in this issue offer a framework of insights about the neuroanatomy of schizophrenia from three learned and creative perspectives. All three articles advance our understanding of schizophrenia from a single locus/specific "lesion" model to more advanced perspectives of neural circuit dysfunction models. Goldman-Rakic and Selemon review their own and others' work on structure-activity relationships of the frontal cortex and related working memory dysfunction. This important but sometimes cloudy and complex area is illuminated by their highly specific, informative research. Jones focuses on thalamic abnormalities hypothetically linked to abnormal oscillations in large arrays of cortical and thalamic neurons, a critically important concept in understanding the functional consequences of abnormal (thalamic) brain structure and function. Graybiel describes her interest in abnormal basal ganglia activity dependent loops that may access the thalamus and set the tone of thalamo-cortical transmission. This view allows for us to understand the "upward" influences on basal ganglia function (and dysfunction) relevant to schizophrenia. These intriguing articles raise a number of issues that await increased data and continued integrating insights. These issues include the need for more information about (1) the developmental timing of lesions or dysfunctions; (2) the extent and regional distribution of abnormalities; (3) the relationship of brain dysfunction to clinical/cognitive abnormalities; and (4) the variable expression of brain abnormalities across the schizophrenia spectrum. These three articles and their authors are at the forefront of our expanding knowledge about the neuroanatomy of schizophrenia and how complex structural and functional deficits are expressed in individuals in the group of schizophrenias. PMID- 9327514 TI - The temporolimbic system theory of paranoid schizophrenia. AB - The hippocampus serves as a funnel for heavily processed sensory information that has converged at the entorhinal cortex. Lesions of the hippocampus do not alter incoming sensory or motor information but, rather, alter their integration with our baggage of emotional experiences and social values. According to Bogerts, such a lesion would be ideally situated to result in laboriously processed sensory information that is out of context to our outside environment. In this regard, Bogerts describes the pathological findings of a patient with a gross delusional disorder. The salient finding at autopsy was a developmental lesion in the left posterior parahippocampal gyrus. Although a number of lesions have been described in the brains of patients with schizophrenia, Bogerts believes that those in the limbic system appear critical to the expression of paranoid symptoms. PMID- 9327515 TI - Functional and anatomical aspects of prefrontal pathology in schizophrenia. AB - Neuropathology is a field that correlates autopsy findings to clinical symptomatology. Since the brain has an inordinate number of parceled regions, each having a different function, it makes more sense to work in an inverse fashion and use clinical findings to establish pathological correlations. In this regard, a lesion in the prefrontal lobes can explain some of the salient findings in schizophrenia, for example, scrambled language, disordered thinking, and abnormal behavior. Recent quantitative cytoarchitectural observations by Goldman Rakic and Selemon sustain such a correlation. By using a computerized image analysis system, these authors have described an abnormally high neuronal density and reduced cortical thickness in many of their patients with schizophrenia. The importance of these findings is discussed in terms of the recent schizophrenia literature. PMID- 9327516 TI - Temporolimbic or transcallosal connections: where is the primary lesion in schizophrenia and what is its nature? AB - A critique of the article by Bogerts on the temporolimbic system theory is presented. Schizophrenia is conceived as arising as a component of the diversity of interhemisphric (callosal) connectivity associated with the evolution of language, a process that occurred through a genetic change (the speciation event) that allowed the hemispheres to develop with a degree of independence. Language and psychosis thus have a common evolutionary origin. The anatomical changes can be considered as a boundary component of the anatomical variation that is characteristic of the species. PMID- 9327517 TI - Functional brain imaging and the neuropathology of schizophrenia. AB - We know much more about the anatomy of the frontal lobes than we do about their functions or how these functions go wrong in schizophrenia. Key areas for brain imaging research that will increase our understanding of frontal function concern (1) long-range functional connectivity and the mechanisms by which one brain region modulates activity in another and (2) the mechanisms underlying specific signs and symptoms associated with schizophrenia. PMID- 9327518 TI - Schizophrenia and disordered neural circuitry. AB - The knowledge of how alterations in neural circuitry relate to the symptoms of schizophrenia may depend on our ability to disentangle a complex cascade of pathological events that affect multiple brain regions. Recent progress in Alzheimer's disease research may provide several useful guiding principles in this pursuit. PMID- 9327519 TI - Discussion of Bogerts' temporolimbic system theory of paranoid schizophrenia. AB - Olney and Farber present their work with N-methyl-D-aspartate (NMDA) antagonists, which are psychotogens, and propose that the structural changes described by Bogerts could be accounted for by a two-stage process. The first stage of the process would occur early in life and would culminate in the selective loss of NMDA-receptor bearing gamma-aminobutyric acid (GABA)ergic neurons and thus render the brain into a NMDA receptor hypofunctional (NRH) state. Such a loss would set the foundation for the second stage in which the neural circuits that have been altered by the loss of these GABAergic interneurons would become activated in late adolescence but would be dysfunctional. Dysfunction of this circuit would lead to the psychopathology of schizophrenia and potentially, if severe enough, to neuronal degeneration. Thus, the changes described by Bogerts could originate partially in early life and partially in adulthood. Based on their animal model, the authors suggest studies that should be carried out in humans. PMID- 9327521 TI - First person account: meaning of psychoses. PMID- 9327520 TI - On localizing schizophrenic neuropathology. AB - Many brain regions and circuits have been implicated in the neuropathology of schizophrenia. Drs. Bogerts and Jones have reviewed the evidence that links the disorder to temporal limbic structures and to frontal-thalamic circuits, respectively. Each article is an important update on what we know about the relevance of these brain regions to schizophrenia. In addition, each article, in summarizing the accumulation of relevant research data, is a testament to the likelihood that these structures play a role in the disease. In light of their compelling arguments, this commentary emphasizes incompleteness in the data and inconsistencies in published findings. The principal weaknesses of the temporal limbic findings are that most have been reported in chronically ill patients and that the only qualitative finding of cytoarchitectural disorganization has not been replicated convincingly. Problems of replication also compromise the interpretation of neuropathological findings in prefrontal cortex and thalamus. Despite the loose ends, I agree with the conclusions of Drs. Bogerts and Jones that brain circuits involving thalamus, prefrontal, and temporolimbic cortices are involved in the basic biology of schizophrenia. PMID- 9327522 TI - DNA vaccines. AB - DNA vaccines use eukaryotic expression vectors to produce immunizing proteins in the vaccinated host. Popular methods of delivery are intramuscular and intradermal saline injections of DNA and gene gun bombardment of skin with DNA coated gold beads. The method of DNA inoculation (gene gun versus intramuscular injection) and the form of the DNA-expressed antigen (cell-associated versus secreted) determine whether T-cell help will be primarily type 1 or type 2. Mechanistically, gene gun-delivered DNA initiates responses by transfected or antigen-bearing epidermal Langerhans cells that move in lymph from bombarded skin to the draining lymph nodes. Following i.m. injections, the functional DNA appears to move as free DNA through blood to the spleen where professional antigen presenting cells initiate responses. Preclinical trials with DNA vaccines have had outstanding success. DNA-based immunizations have provided protection against viral, bacterial and parasitic diseases, modulated the effects of autoimmune and allergic disease, and provided some hope for the control of cancer. PMID- 9327523 TI - IL-12 as an adjuvant for cell-mediated immunity. AB - Recent studies have shown that IL-12 initiates the development of cell-mediated immunity following infection with certain infectious agents. This attribute of IL 12 makes it a potentially effective adjuvant for vaccines. This review summarizes our findings in the Leishmania system, outlining the role of endogenous IL-12 in resistance to L. major and the use of exogenous IL-12 in a vaccine against L. major, and also describes other examples in which IL-12 has been shown to be effective as an adjuvant at inducing protective immunity. The potential use of IL 12 as an adjuvant in therapeutic vaccines is also discussed. PMID- 9327524 TI - The immunotargeting approach to adjuvant-independent subunit vaccine design. AB - Recently there has been considerable interest in exploring adjuvant-independent approaches to the enhancement of immunogenicity. One strategy which has emerged in response to this need is the immunotargeting approach to subunit vaccine design. Immunotargeting involves the conjugation of non-self antigens to monoclonal antibodies specific for cell surface determinants (e.g. class II MHC) on antigen presenting cells in vivo. Saline injections of these immunoconjugates have been shown to promote significant IgG responses to the delivered antigens in a number of different animal model studies. Oral and nasal immunizations in mice with anti-class II MHC immunoconjugates induce both secretory IgA and systemic IgG responses. Recombinant anticlass II MHC antibodies with defined B-cell and T cell epitopes incorporated into their primary sequence have recently been used to induce epitope specific antibody responses in macaques. Thus the potential now exists for the rational design of adjuvant-independent human vaccine candidates based on a recombinant immunotargeting anti-body framework. PMID- 9327525 TI - Challenges for vaccination against sexually-transmitted diseases: induction and long-term maintenance of mucosal immune responses in the female genital tract. AB - The ability to develop vaccines capable of preventing or protecting against sexually transmitted viruses, such as herpes simplex virus (HSV) and HIV is likely to depend on the induction of long-term mucosal immune responses. We have utilized a recombinant adenovirus capable of expressing HSV glycoprotein B (AdgB8) to demonstrate that intranasal (i.n.) immunization induces HSVgB-specific IgG and IgA antibodies in the serum and vaginal washes of mice. In contrast, systemic immunization only resulted in IgG antibodies in vaginal fluids. We also observed that although i.n. and i.p. AdgB8 immunization induced short-term CTL responses, only i.n. immunized mice maintained long-term CTL responses in the genital-associated lymphoid tissues. When compared to systemic immunization, mice immunized i.n. with the same dose of AdgB8 were better protected for a longer time period from a lethal intravaginal HSV-2 challenge. Protection occurred despite the fact that mice were initially infected (i.e. not sterile immunity) and the enhanced survival occurring in i.n. immunized mice correlated with the presence of HSVgB-specific IgA antibody-secreting cells in the genital tissues and with memory CTL, recall responses. Together, these results indicate that mucosal immunization is important for the induction and long-term maintenance of mucosal immune responses. PMID- 9327526 TI - Priming of T cells by heat shock protein-peptide complexes as the basis of tumor vaccines. AB - A number of heat shock proteins, or stress proteins, have been shown functionally as potent 'tumor rejection antigens', despite the fact that there are no single tumor-specific mutations in these proteins. Current studies have solved many aspects of this puzzle biochemically and immunologically. The proposal that heat shock proteins are not antigenic per se, but chaperone and present antigenic peptides to MHC class I molecules for recognition by T lymphocytes, has been mechanistically defined. This significance of this discovery is elaborated in the context of the importance of MHC molecules. The immunological principle and the practical vaccination strategy of the two major mammalian heat shock proteins, gp96/grp94 and hsp70 are discussed. It is also argued that extracellular heat shock proteins signal cell stress and tissue damage, and are involved in presenting peptides to the MHC class I pathway through professional antigen presenting cells. These results and ideas form the basis of a new generation of T cell vaccines against tumors and intracellular organisms. PMID- 9327527 TI - Activation or frustration of anti-tumor responses by T-cell-based immune modulation. AB - Many types of tumors (e.g. virus-induced tumors, melanomas, tumors over expressing certain oncogenes) often express antigens that can induce T-cell mediated tumor-specific immune responses. Nonetheless, many such tumors manage to circumvent the induction of an effective anti-tumor T-cell response, as is apparent from the many tumor-bearing patients. Therefore, optimally designed vaccination protocols may evoke a more powerful and competent T-cell-mediated anti-tumor response, allowing the host to effectively deal with at least some cancers. These vaccination approaches might include immunization with whole (tumor) cell-based vaccines, entire tumor antigens or selected T-cell epitopes derived from tumor antigens. This survey is an update of several anti-tumor vaccination approaches employed, and on novel possibilities to target the anti tumor immune response to preselected tumor-derived T-cell epitopes. PMID- 9327528 TI - Influence of strength training on sprint running performance. Current findings and implications for training. AB - Today, it is generally accepted that sprint performance, like endurance performance, can improve considerably with training. Strength training, especially, plays a key role in this process. Sprint performance will be viewed multidimensionally as an initial acceleration phase (0 to 10 m), a phase of maximum running speed (36 to 100 m) and a transition phase in between. Immediately following the start action, the powerful extensions of the hip, knee and ankle joints are the main accelerators of body mass. However, the hamstrings, the m. adductor magnus and the m. gluteus maximus are considered to make the most important contribution in producing the highest levels of speed. Different training methods are proposed to improve the power output of these muscles. Some of them aim for hypertrophy and others for specific adaptations of the nervous system. This includes general (hypertrophy and neuronal activation), velocity specific (speed-strength) and movement specific (sprint associated exercises) strength training. In developing training strategies, the coach has to keep in mind that strength, power and speed are inherently related to one another, because they are all the output of the same functional systems. As heavy resistance training results in a fibre type IIb into fibre type IIa conversion, the coach has to aim for an optimal balance between sprint specific and nonspecific training components. To achieve this they must take into consideration the specific strength training demands of each individual, based on performance capacity in each specific phase of the sprint. PMID- 9327529 TI - An overview of some definitional issues for sports injury surveillance. AB - Injury surveillance is the ongoing collection of data describing the occurrence of, and factors associated with, injury. The success of any sports injury surveillance system and its wide scale applicability is dependent upon valid and reliable definitions of sports injury, injury severity and sports participation. Published sports injury reports are often difficult to interpret and compare with other published data because of different data collection and/or analysis methods. Standardised data collection methodologies including definitions are crucial for improving the comparability and interpretation of published data. Attention needs to be directed towards the definition of both risk and exposure factors since the validity and usefulness of the outcomes of research activities, data collection and surveillance systems rely on these. International consensus on appropriate definitions would greatly assist the collection of comparable and reliable sports injury data. Standardised definitions are also needed to answer questions such as: 'what is a sport? When should an activity be considered to be recreational rather than sport? Who is a sports participant? How should sports participation be measured? What is a meaningful measure of exposure to injury risk? What is a sports injury? How should sports injury severity be measured? How severe must an injury be before it should be considered to be a sports injury for surveillance purposes?' Agreed definitions and answers to these questions are essential before injury surveillance is established. Sports injury data is needed to guide injury prevention activities, to set and monitor sports safety policies and interventions, and as the basis of sports injury prevention research. All sports injury surveillance systems should therefore collect information about the epidemiology of sports injuries and their outcomes in a form that is of relevance across a broad range of potential users of the data. PMID- 9327530 TI - Sports injury surveillance systems. 'One size fits all'? AB - Sport is beneficial to health, but may also cause injuries. Therefore there is a need for sports injury prevention. Sports injury prevention should be based on the outcome of scientific research and should be part of the 'sequence of prevention'. In applying the 'sequence of prevention', first the incidence and severity of the sports injury problem need to be established. Secondly the aetiology and the mechanism of sports injuries need to be identified. Only based on this information can preventative measures be introduced, which must subsequently be evaluated for effectiveness. The principle of the 'sequence of prevention' cannot be applied without proper sports injury surveillance. This paper addresses the question of whether one uniform sports injury surveillance system can be used to cover all aspects of sports injury research at all stages of the 'sequence of prevention'. It is argued that a general sports injury surveillance system is useful for answering questions about the incidence and severity of the sports injury problem in various subsets of a population. It can also be used for time trend studies. If the purpose of injury surveillance is to identify the aetiology or the effectiveness of preventative measures, then sports injury surveillance should be tailored to the specific sports situation. Sports injury surveillance systems are not useful in identifying the mechanism of injury. PMID- 9327531 TI - Sports injury surveillance has everything to do with sports medicine. AB - Injury is the most under-recognised major public health problem facing the world community. In addition to being an enormous public health issue, injuries continue to usurp our limited healthcare financial resources. It therefore becomes imperative that comprehensive injury occurrence data systems be developed in order to: (i) identify risk factors and types of injuries; (ii) help determine the effectiveness of preventative intervention; and (iii) delineate the costs of injury in order to develop financial resource planning. PMID- 9327532 TI - Exposure data. Why are they needed? AB - Aspects of different approaches to study design in the epidemiology of sports injuries are presented, identifying 4 major types: (i) the clinical case series; (ii) the community-based survey; (iii) studies on specific sports or diagnoses without exposure data; and (iv) studies on specific sports or diagnoses with exposure data. The advantages and disadvantages of these concepts are discussed. It is concluded that to optimise preventive efforts and improve the comparability of studies in sports injury epidemiology, there is not only a need for consensus on definitional issues, but also for an agreement on the methodology. Attention to exposure issues is a crucial component of this. PMID- 9327534 TI - Sports injury surveillance and protective equipment. AB - Protective equipment is adopted in the hope of reducing the incidence and severity of injuries. To objectively assess the effectiveness of such equipment, injury data is required prior to and after the introduction of this countermeasure. In many cases, there has been no appropriate evaluation of the countermeasure. Pre- and postintervention data is vital to the protective equipment developer. Such information may be obtained from injury surveillance systems. Other information which is just as vital but not obtainable from such a system is injury tolerance levels. Additional information from injury surveillance such as the type of protective equipment worn, its condition prior to and after impact, and a description of the event leading to injury would be most valuable. PMID- 9327533 TI - The severity of sports injuries. AB - In order to assess the extent of the sports injury problem, it is necessary to identify both the incidence and severity of sports injuries. In the literature, the severity of sports injuries is usually described on the basis of 6 criteria: (i) nature of sports injury; (ii) duration and nature of treatment; (iii) sporting time lost; (iv) working time lost; (v) permanent damage; and (vi) monetary cost. It is important to include information on the severity of injuries in a sports injury registration system. This kind of information will help to set targets for preventive strategies. The more severe the injuries sustained, the higher the priority will be to prevent these injuries regardless of the injury incidence. When assessing injury severity one should take into account that, in order to enhance the comparability of research data, uniform definitions are also important in this area of sports injury surveillance. This paper briefly discusses the 6 criteria used to describe the severity of sports injuries. PMID- 9327535 TI - Athletic injury reporting. Development of universal systems. AB - There are numerous athletic injury reporting systems currently in place. In order for our understanding of athletic injury epidemiology to advance, we must be able to compare data from divergent sources. This paper provides a review of existing athletic injury reporting systems in North America. The epidemiological designs employed in these systems are outlined, along with a description of the strengths and weaknesses of each approach to reporting. The differences between the case series and cohort methods are delineated and the importance of injury definition, sources of error, denominator data and exposure estimation are discussed within this context. Four recommendations are then offered to assist in moving toward more universal systems for athletic injury reporting. First, comparability of data between systems should be maximised through clear indication of the reporting system design and the methods of data collection. Secondly, an exact definition should be given as to what constitutes a reportable event ('injury'). Thirdly, whenever possible, outcome information should be collected on each reported event so that an injury definition may be applied at the time of data analysis. Lastly, any limitations or sources of error should be acknowledged. PMID- 9327536 TI - Rotator cuff pathology in athletes. AB - The rotator cuff is the primary dynamic stabiliser of the glenohumeral joint and is placed under significant stress during overhead and contact sports. Mechanisms of injury include repetitive microtrauma, usually seen in the athlete involved in overhand sports, and macrotrauma associated with contact sports. Rotator cuff injury and dysfunction in the overhand athlete may be classified based on aetiology as primary impingement, primary tensile overload, and secondary impingement and tensile overload resulting from glenohumeral instability. A thorough history and physical examination are paramount in the evaluation, classification and treatment planning of the athlete with rotator cuff pathology. Imaging studies are a helpful adjunct to the history and physical. Athletes with primary impingement are usually middle aged or older and often have chronic shoulder pain and sometimes weakness associated with overhand sporting activities. Night pain is typical of full thickness rotator cuff tears. Impingement signs are positive and strength of elevation and external rotation are often limited. They usually respond to a nonoperative rehabilitation programme centred on decreasing inflammation, restoring range of motion and strengthening the rotator cuff and scapular stabilisers. Depending on the degree of cuff pathology, acromioplasty, debridement of partial cuff tears, and repair of full thickness tears are usually successful in those who fail a rehabilitation programme. Overhand athletes with cuff pathology secondary to subtle anterior instability are usually young and complain of pain and decreased throwing velocity. Instability may be so subtle that it is only detectable through a positive relocation test on examination. The majority of these athletes do not have a Bankart lesion on magnetic resonance imaging or arthroscopic examination. Arthroscopic examination usually demonstrates anterior capsular laxity (positive 'drive-through' sign), as well as superior-posterior labral and cuff injury typical of internal impingement. If rehabilitation alone is not successful, a capsulolabral repair followed by rehabilitation may allow the athlete to return to their previous level of competition. The athlete with an acute episode of macrotrauma to the shoulder resulting in cuff pathology usually presents with pain, limited active elevation and a positive 'shrug sign'. Arthroscopy and debridement of thickened, inflamed or scarred subacromial bursa with cuff repair or debridement as indicated is usually successful in those who do not respond to a rehabilitation programme. PMID- 9327540 TI - A model for the combined effects of temperature and salt concentration on growth rate of food spoilage molds. AB - We modeled mold growth on a solid culture medium at various temperatures and NaCl concentrations by using five common food spoilage molds (Penicillium roqueforti, Trichoderma harzianum, Paecilomyces variotii, Aspergillus niger, and Emericella nidulans). For the description of the growth rate (expressed as the increase in colony diameter per unit of time) as a function of temperature and NaCl concentration, a six-parameter model has been developed. The model combines either the Rosso-type or the Ratkowsky-type temperature dependence with the NaCl concentration dependence derived from the relationship between the growth rate and square root of (1 - water activity), as proposed by Gibson and coworkers (A. M. Gibson, J. Baranyi, J. I. Pitt, M. J. Eyles, and T. A. Roberts, Int. J. Food Microbiol. 23:419-431, 1994). The model will be of use to food microbiologists whose aim is to predict the likelihood of fungal spoilage. PMID- 9327537 TI - Inhibition and facilitation of nucleic acid amplification. PMID- 9327538 TI - Isolation and overexpression of a gene encoding an extracellular beta-(1,3-1,4) glucanase from Streptococcus bovis JB1. AB - Streptococcus bovis JB1 was found to produce a 25-kDa extracellular enzyme active against beta-(1,3-1,4)-glucans. A gene was isolated encoding a specific beta-(1,3 1,4)-glucanase that corresponds to this size and belongs to glycoside hydrolase family 16. A 4- to 10-fold increase in supernatant beta-glucanase activity was obtained when the cloned beta-glucanase gene was reintroduced into S. bovis JB1 by use of constructs based on the plasmid vector pTRW10 or pIL253. The beta-(1,3 1,4)-glucanase gene was also expressed upon introduction of the pTRW10 construct pTRWL1R into Lactococcus lactis IL2661 and Enterococcus faecalis JH2-SS, although extracellular activity was 8- to 50-fold lower than that in S. bovis JB1. The beta-(1,3-1,4)-glucanase purified from the culture supernatant of S. bovis JB1 carrying pTRWL1R showed a K(m) of 2.8 mg per ml and a Vmax of 338 mumol of glucose equivalents per min per mg of protein with barley beta-glucan as the substrate. The S. bovis beta-(1,3-1,4)-glucanase may contribute to the ability of this bacterium to utilize starch by degrading structural polysaccharides present in endosperm cell walls. PMID- 9327541 TI - Purification and characterization of phenylacetaldehyde reductase from a styrene assimilating Corynebacterium strain, ST-10. AB - A novel phenylacetaldehyde reductase was purified about 50-fold to homogeneity from Corynebacterium sp. strain ST-10, which can assimilate gaseous styrene as the sole carbon and energy source. The enzyme was inductively synthesized when grown on gaseous styrene and had an important role in styrene metabolism in vivo. The enzyme had a molecular weight of 155,000 and was composed of four identical subunits (molecular weight, 42,000). The enzyme catalyzed the reduction of not only phenylacetaldehyde but also various aldehydes and ketones; however, it did not catalyze the reverse reaction, the dehydrogenation of 2-phenylethanol. The enzyme required NADH as a cofactor and showed no activity with NADPH; therefore, it was defined as an NADH-dependent phenylacetaldehyde reductase. The enzyme stereospecifically produced (S)-(-)-1-phenylethanol from acetophenone; therefore, it would be useful as a biocatalyst. PMID- 9327543 TI - Streptococcus thermophilus and its biosurfactants inhibit adhesion by Candida spp. on silicone rubber. AB - The adhesion of yeasts, two Candida albicans and two Candida tropicalis strains isolated from naturally colonized voice prostheses, to silicone rubber with and without a salivary conditioning film in the absence and presence of adhering Streptococcus thermophilus B, a biosurfactant-releasing dairy isolate, was studied. Coverage of 1 to 4% of the surface of silicone rubber substrata with adhering S. thermophilus B gave significant reductions in the initial yeast adhesion regardless of the presence of a conditioning film. Mechanistically, this interference in yeast adhesion by S. thermophilus B was not due to direct physical effects but to biosurfactant release by the adhering bacteria, because experiments with S. thermophilus B cells that had released their biosurfactants prior to adhesion to silicone rubber and competition with yeasts did not show interference with initial yeast adhesion. The amounts of biosurfactants released were highest for mid-exponential- and early-stationary-phase bacteria (37 mg.g of cells-1 [dry weight]), but biosurfactants released by stationary-phase bacteria (14 mg.g of cells-1 [dry weight]) were the most surface active. The crude biosurfactants released were mixtures of various components, with a glycolipid like component being the most surface active. A lipid-enriched biosurfactant fraction reduced the surface tension of an aqueous solution to about 35 mJ.m-2 at a concentration of only 0.5 mg.ml-1. The amount of biosurfactant released per S. thermophilus B cell was estimated to be sufficient to cover approximately 12 times the area of the cross section of the bacterium, making biosurfactant release a powerful defense weapon in the postadhesion competition of the bacterium with microorganisms such as yeasts. Preadsorption of biosurfactants to the silicone rubber prior to allowing yeasts to adhere was as effective against C. albicans GB 1/2 adhesion as covering 1 to 2% of the silicone rubber surface with adhering S. thermophilus B, but a preadsorbed biosurfactant layer was less effective against C. tropicalis GB 9/9. PMID- 9327542 TI - Increased species diversity and extended habitat range of sulfur-oxidizing Thiomicrospira spp. AB - We combined traditional cultivation methods and new molecular techniques to study the diversity and habitat range of bacteria of the genus Thiomicrospira. Specific primers were designed and used in the PCR to amplify the 16S ribosomal DNA (rDNA) of Thiomicrospira spp. and thus detect the presence of these bacteria in environmental samples and enrichment cultures. By using this genus-specific PCR, we were able to amplify 722-bp-long 16S rDNA fragments from different saltwater habitats as well as from a freshwater ecosystem. Furthermore, we were able to isolate most of these bacteria in pure culture by using enrichment cultures for chemolithoautotrophic sulfur-oxidizing bacteria. With denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA fragments followed by hybridization analysis with one of the primers as a genus-specific probe, it was possible to monitor the success of isolation. The combined approach resulted in the isolation of several chemolithoautotrophic bacteria from different habitats: e.g., a coastal sediment along the coast of Chile, a microbial mat of the hypersaline pond Solar Lake (Sinai, Egypt), and the saline spring Artern (Thuringia, Germany). In addition, four different isolates were obtained from sediment and water samples taken at Jadebusen, which is part of the German Waddensea. Comparative analysis of the nearly complete 16S rRNA sequences of these isolates indicated several new species, all grouping with the Thiomicrospira species of the gamma subdivision of the class Proteobacteria. A freshwater Thiomicrospira species could not be isolated, but sequence analysis of the PCR product obtained after amplification of the environmental DNA with the Thiomicrospira-specific primers revealed its phylogenetic affiliation. The study indicates an increased species diversity of Thiomicrospira and the ubiquity of this sulfur-oxidizing bacterium in habitats with reduced sulfur compounds. PMID- 9327544 TI - The freeze-thaw stress response of the yeast Saccharomyces cerevisiae is growth phase specific and is controlled by nutritional state via the RAS-cyclic AMP signal transduction pathway. AB - The ability of cells to survive freezing and thawing is expected to depend on the physiological conditions experienced prior to freezing. We examined factors affecting yeast cell survival during freeze-thaw stress, including those associated with growth phase, requirement for mitochondrial functions, and prior stress treatment(s), and the role played by relevant signal transduction pathways. The yeast Saccharomyces cerevisiae was frozen at -20 degrees C for 2 h (cooling rate, less than 4 degrees C min-1) and thawed on ice for 40 min. Supercooling occurred without reducing cell survival and was followed by freezing. Loss of viability was proportional to the freezing duration, indicating that freezing is the main determinant of freeze-thaw damage. Regardless of the carbon source used, the wild-type strain and an isogenic petite mutant ([rho 0]) showed the same pattern of freeze-thaw tolerance throughout growth, i.e., high resistance during lag phase and low resistance during log phase, indicating that the response to freeze-thaw stress is growth phase specific and not controlled by glucose repression. In addition, respiratory ability and functional mitochondria are necessary to confer full resistance to freeze-thaw stress. Both nitrogen and carbon source starvation led to freeze-thaw tolerance. The use of strains affected in the RAS-cyclic AMP (RAS-cAMP) pathway or supplementation of an rca1 mutant (defective in the cAMP phosphodiesterase gene) with cAMP showed that the freeze-thaw response of yeast is under the control of the RAS-cAMP pathway. Yeast did not adapt to freeze-thaw stress following repeated freeze-thaw treatment with or without a recovery period between freeze-thaw cycles, nor could it adapt following pretreatment by cold shock. However, freeze-thaw tolerance of yeast cells was induced during fermentative and respiratory growth by pretreatment with H2O2, cycloheximide, mild heat shock, or NaCl, indicating that cross protection between freeze-thaw stress and a limited number of other types of stress exists. PMID- 9327545 TI - Novel detoxification of the trichothecene mycotoxin deoxynivalenol by a soil bacterium isolated by enrichment culture. AB - A mixed microbial culture capable of metabolizing deoxynivalenol was obtained from soil samples by an enrichment culture procedure. A bacterium (strain E3-39) isolated from the enrichment culture completely removed exogenously supplied deoxynivalenol from culture medium after incubation for 1 day. On the basis of morphological, physiological, and phylogenetic studies, strain E3-39 was classified as a bacterium belonging to the Agrobacterium-Rhizobium group. Thin layer chromatographic analysis indicated the presence of one major and two minor metabolites of deoxynivalenol in ethyl acetate extracts of the E3-39 culture filtrates. The main metabolite was identified as 3-keto-4-deoxynivalenol by mass spectroscopy and 1H and 13C nuclear magnetic resonance analysis. The immunosuppressive toxicity of 3-keto-4-deoxynivalenol was evaluated by means of a bioassay based on the mitogen-induced and mitogen-free proliferations of mouse spleen lymphocytes. This compound exhibited a remarkably decreased (to less than one tenth) immunosuppressive toxicity relative to deoxynivalenol, indicating that the 3-OH group in deoxynivalenol is likely to be involved in exerting its immunosuppressive toxicity. Strain E3-39 was also capable of transforming 3 acetyldeoxynivalenol but not nivalenol and fusarenon-X. PMID- 9327546 TI - Moderate concentrations of ethanol inhibit endocytosis of the yeast maltose transporter. AB - The maltose transporter in Saccharomyces cerevisiae is degraded in the vacuole after internalization by endocytosis upon nitrogen starvation in the presence of a fermentable substrate. This degradation, known as catabolite inactivation, is inhibited by the presence of moderate concentrations (2 to 6%, vol/vol) of ethanol. We have investigated the mechanism of this inactivation and have found that it is due to the inhibition of the internalization of the transporter by endocytosis. The results also indicate that this inhibition is due to alterations produced by ethanol in the organization of the plasma membrane which also affects to endocytosis of other plasma membrane proteins. Apparently, endocytosis is particularly sensitive to these alterations compared with other processes occurring at the plasma membrane. PMID- 9327547 TI - Assessment of a dye permeability assay for determination of inactivation rates of Cryptosporidium parvum oocysts. AB - The ability to determine inactivation rates of Cryptosporidium parvum oocysts in environmental samples is critical for assessing the public health hazard of this gastrointestinal parasite in watersheds. We compared a dye permeability assay, which tests the differential uptake of the fluorochromes 4'-6-diamidino-2 phenylindole (DAPI) and propidium iodide (PI) by the oocysts (A. T. Campbell, L. J. Robertson, and H. V. Smith, Appl. Environ. Microbiol. 58:3488-3493, 1992), with an in vitro excystation assay, which tests their ability to excyst and, thus, their metabolic potential and potential for infectivity (J.B. Rose, H. Darbin, and C.P. Gerba, Water Sci. Technol. 20:271-276, 1988). Formaldehyde-fixed (killed) oocysts and untreated oocysts were permeabilized with sodium hypochlorite and subjected to both assays. The results of the dye permeability assays were the same, while the excystation assay showed that no excystation occurred in formaldehyde-fixed oocysts. This confirmed that oocyst wall permeability, rather than metabolic activity potential, was the basis of the dye permeability viability assessment. A previously developed protocol (L. J. Anguish and W. C. Ghiorse, Appl. Environ. Microbiol. 63:724-733, 1997) for determining viability of oocysts in soil and sediment was used to examine further the use of oocyst permeability status as an indicator of oocyst viability in fecal material stored at 4 degrees C and in water at various temperatures. Most of the oocysts in fresh calf feces were found to be impermeable to the fluorochromes. They were also capable of excystation, as indicated by the in vitro excystation assay, and were infective, as indicated by a standard mouse infectivity assay. The dye permeability assay further showed that an increase in the intermediate population of oocysts permeable to DAPI but not to PI occurred over time. There was also a steady population of oocysts permeable to both dyes. Further experiments with purified oocysts suspended in distilled water showed that the shift in oocyst populations from impermeable to partially permeable to fully permeable was accelerated at temperatures above 4 degrees C. This sequence of oocyst permeability changes was taken as an indicator of the oocyst inactivation pathway. Using the dye permeability results, inactivation rates of oocysts in two fecal pools stored in the dark at 4 degrees C for 410 and 259 days were estimated to be 0.0040 and 0.0056 oocyst day-1, respectively. The excystation assay gave similar inactivation rates of 0.0046 and 0.0079 oocyst day-1. These results demonstrate the utility of the dye permeability assay as an indicator of potential viability and infectivity of oocysts, especially when combined with improved microscopic methods for detection of oocysts in soil, turbid water, and sediments. PMID- 9327548 TI - Cloning and sequence analysis of a novel hemolysin gene (vllY) from Vibrio vulnificus. AB - A gene (vllY) encoding a novel hemolysin of Vibrio vulnificus CKM-1 has been cloned and sequenced. When the vllY gene was expressed in minicells, a unique peptide of approximately 40 kDa was identified. Subcellular fractionation of Escherichia coli cells carrying the vllY gene indicated that the VllY protein was distributed in both the cytoplasmic and the periplasmic fractions, with the notable ability to appear in the latter compartment. Nucleotide sequence analysis predicted a single open reading frame of 1,071 bp encoding a 357-amino acid polypeptide with an estimated pI of 5.02. The deduced amino acid sequence of VllY showed high similarity to the sequence of legiolysin, responsible for hemolysis, pigment production, and fluorescence in Legionella pneumophila. The enzyme also exhibited sequence homology to the MelA protein sequence of Shewanella colwelliana and the sequences of 4-hydroxyphenylpyruvate dioxygenase family proteins from various organisms. PCR screening and Southern blotting of V. vulnificus strains revealed that all of the 41 V. vulnificus clinical isolates contained vllY-like genes. PMID- 9327549 TI - Detection and characterization of fungal infections of Ammophila arenaria (marram grass) roots by denaturing gradient gel electrophoresis of specifically amplified 18s rDNA. AB - Marram grass (Ammophila arenaria L.), a sand-stabilizing plant species in coastal dune areas, is affected by a specific pathosystem thought to include both plant pathogenic fungi and nematodes. To study the fungal component of this pathosystem, we developed a method for the cultivation-independent detection and characterization of fungi infecting plant roots based on denaturing gradient gel electrophoresis (DGGE) of specifically amplified DNA fragments coding for 18S rRNA (rDNA). A nested PCR strategy was employed to amplify a 569-bp region of the 18S rRNA gene, with the addition of a 36-bp GC clamp, from fungal isolates, from roots of test plants infected in the laboratory, and from field samples of marram grass roots from both healthy and degenerating stands from coastal dunes in The Netherlands. PCR products from fungal isolates were subjected to DGGE to examine the variation seen both between different fungal taxa and within a single species. DGGE of the 18S rDNA fragments could resolve species differences from fungi used in this study yet was unable to discriminate between strains of a single species. The 18S rRNA genes from 20 isolates of fungal species previously recovered from A. arenaria roots were cloned and partially sequenced to aid in the interpretation of DGGE data. DGGE patterns recovered from laboratory plants showed that this technique could reliably identify known plant-infecting fungi. Amplification products from field A. arenaria roots also were analyzed by DGGE, and the major bands were excised, reamplified, sequenced, and subjected to phylogenetic analysis. Some recovered 18S rDNA sequences allowed for phylogenetic placement to the genus level, whereas other sequences were not closely related to known fungal 18S rDNA sequences. The molecular data presented here reveal fungal diversity not detected in previous culture-based surveys. PMID- 9327550 TI - Trichloroethylene biodegradation by mesophilic and psychrophilic ammonia oxidizers and methanotrophs in groundwater microcosms. AB - This study investigated the efficiency of methane and ammonium for stimulating trichloroethylene (TCE) biodegradation in groundwater microcosms (flasks and batch exchange columns) at a psychrophilic temperature (12 degrees C) typical of shallow aquifers in the northern United States or a mesophilic temperature (24 degrees C) representative of most laboratory experiments. After 140 days, TCE biodegradation rates by ammonia oxidizers and methanotrophs in mesophilic flask microcosms were similar (8 to 10 nmol day-1), but [14C]TCE mineralization (biodegradation to 14CO2) by ammonia oxidizers was significantly greater than that by methanotrophs (63 versus 53%). Under psychrophilic conditions, [14C]TCE mineralization in flask systems by ammonia oxidizers and methanotrophs was reduced to 12 and 5%, respectively. In mesophilic batch exchange columns, average TCE biodegradation rates for methanotrophs (900 nmol liter-1 day-1) were not significantly different from those of ammonia oxidizers (775 nmol liter-1 day-1). Psychrophilic TCE biodegradation rates in the columns were similar with both biostimulants and averaged 145 nmol liter-1 day-1. Methanotroph biostimulation was most adversely affected by low temperatures. At 12 degrees C, the biodegradation efficiencies (TCE degradation normalized to microbial activity) of methanotrophs and ammonia oxidizers decreased by factors of 2.6 and 1.6, respectively, relative to their biodegradation efficiencies at 24 degrees C. Collectively, these experiments demonstrated that in situ bioremediation of TCE is feasible at the psychrophilic temperatures common in surficial aquifers in the northern United States and that for such applications biostimulation of ammonia oxidizers could be more effective than has been previously reported. PMID- 9327551 TI - Evaluation of dietary influences on Escherichia coli O157:H7 shedding by sheep. AB - The effect of diet, an abrupt diet change, and fasting on the shedding of Escherichia coli O157:H7 was investigated with experimentally inoculated sheep as a ruminant model. Sheep were fed a grass hay diet (G), which was low in protein and digestible energy and high in fiber, or a mixture of corn and pelleted alfalfa (C), which was high in protein and digestible energy and low in fiber. After a single oral inoculation of E. coli O157:H7, all the animals shed fecal E. coli O157:H7. However, sheep that were fed G shed the bacterium almost twice as long as, and in larger numbers than, did sheep that were fed C. The number of culture-positive animals increased after the diet was abruptly changed from C to G and decreased with the opposite change (G to C). A 24-h fast did not influence E. coli O157:H7 shedding. Horizontal transmission of infection between animals occurred. Recent shedding of E. coli O157:H7 did not affect recolonization with E. coli O157:H7. The findings presented in this study indicate that preharvest control of diet may reduce the risk of E. coli O157:H7-positive animals entering the food chain. PMID- 9327552 TI - Critical role of anteiso-C15:0 fatty acid in the growth of Listeria monocytogenes at low temperatures. AB - Listeria monocytogenes is a food-borne pathogen capable of growth at refrigeration temperatures. Membrane lipid fatty acids are major determinants of a sufficiently fluid membrane state to allow growth at low temperatures. L. monocytogenes was characterized by a fatty acid profile dominated to an unusual extent (> 95%) by branched-chain fatty acids, with the major fatty acids being anteiso-C15:0, anteiso-C17:0, and iso-C15:0 in cultures grown in complex or defined media at 37 degrees C. Determination of the fatty acid composition of L. monocytogenes 10403S and SLCC 53 grown over the temperature range 45 to 5 degrees C revealed two modes of adaptation of fatty acid composition to lower growth temperatures: (i) shortening of fatty acid chain length and (ii) alteration of branching from iso to anteiso. Two transposon Tn917-induced cold-sensitive mutants incapable of growth at low temperatures had dramatically altered fatty acid compositions with low levels of i-C15:0, a-C15:0, and a-C17:0 and high levels of i-C14:0, C14:0, i-C16:0, and C16:0. The levels of a-C15:0 and a-C17:0 and the ability to grow at low temperatures were restored by supplementing media with 2-methylbutyric acid, presumably because it acted as a precursor of methylbutyryl coenzyme A, the primer for synthesis of anteiso odd-numbered fatty acids. When mid-exponential-phase 10403S cells grown at 37 degrees C were temperature down-shocked to 5 degrees C they were able, for the most part, to reinitiate growth before the membrane fatty acid composition had reset to a composition more typical for low-temperature growth. No obvious evidence was found for a role for fatty acid unsaturation in adaptation of L. monocytogenes to cold temperature. The switch to a fatty acid profile dominated by a-C15:0 at low temperatures and the association of cold sensitivity with deficiency of a-C15:0 focus attention on the critical role of this fatty acid in growth of L. monocytogenes in the cold, presumably through its physical properties and their effects, in maintaining a fluid, liquid-crystalline state of the membrane lipids. PMID- 9327553 TI - Seasonal changes in the relative abundance of uncultivated sulfate-reducing bacteria in a salt marsh sediment and in the rhizosphere of Spartina alterniflora. AB - Phylogenetic diversity and community composition of sulfate-reducing bacteria in a salt marsh sediment and in the rhizosphere of Spartina alterniflora were investigated. Uncultivated Desulfobacteriaceae family-related phylotypes were studied by selectively amplifying 16S rRNA gene fragments from DNA extracted from salt marsh rhizosphere samples. Two novel phylotypes were retrieved from rhizosphere samples, with A01 having 89.1% sequence similarity with Desulfococcus multivorans and 4D19 having 96.3% sequence similarity with Desulfosarcina variabilis. Additionally, six sequences that were extremely closely related to Desulfococcus multivorans (99% sequence similarity) were found. Reference RNAs containing sequences identical to corresponding cloned regions of A01 or 4D19 16S rRNA were synthesized via in vitro transcription and were used in subsequent quantitative membrane hybridization experiments. Oligonucleotide probes A01-183 and 4D19-189 were designed to specifically target these two novel phylotypes and were tested for target specificity against synthesized RNA and reference RNAs extracted from pure cultures. The newly designed probes were then used, together with eubacterial probes, to determine the relative abundances of the novel phylotypes in the salt marsh sediment and the rhizosphere. Mean relative abundances of A01-183 and 4D19-189 targets were 7.5 and 3.4%, respectively, suggesting that the target organisms of A01-183 and, to a lesser extent, of 4D19 189 play an important role in the salt marsh sediment and the Spartina rhizosphere. A seasonal trend of increased A01 relative abundance during the period of vegetative plant growth was evident, suggesting a close interaction between A01 and S. alterniflora. PMID- 9327555 TI - Efficacy of vaporized hydrogen peroxide against exotic animal viruses. AB - The efficacy of vapor-phase hydrogen peroxide in a pass-through box for the decontamination of equipment and inanimate materials potentially contaminated with exotic animal viruses was evaluated. Tests were conducted with a variety of viral agents, which included representatives of several virus families (Orthomyxoviridae, Reoviridae, Flaviviridae, Paramyxoviridae, Herpesviridae, Picornaviridae, Caliciviridae, and Rhabdoviridae) from both avian and mammalian species, with particular emphasis on animal viruses exotic to Canada. The effects of the gas on a variety of laboratory equipment were also studied. Virus suspensions in cell culture media, egg fluid, or blood were dried onto glass and stainless steel. Virus viability was assessed after exposure to vaporphase hydrogen peroxide for 30 min. For all viruses tested and under all conditions (except one), the decontamination process reduced the virus titer to 0 embryo lethal doses for the avian viruses (avian influenza and Newcastle disease viruses) or less than 10 tissue culture infective doses for the mammalian viruses (African swine fever, bluetongue, hog cholera, pseudorabies, swine vesicular disease, vesicular exanthema, and vesicular stomatitis viruses). The laboratory equipment exposed to the gas appeared to suffer no adverse effects. Vaporphase hydrogen peroxide decontamination can be recommended as a safe and efficacious way of removing potentially virus-contaminated objects from biocontainment level III laboratories in which exotic animal disease virus agents are handled. PMID- 9327554 TI - Genes encoding two different beta-glucosidases of Thermoanaerobacter brockii are clustered in a common operon. AB - A 5.9-kb fragment of chromosomal DNA coding for beta-glucosidase activity of the thermophilic anaerobe Thermoanaerobacter brockii was sequenced. Two genes, cglT and xglS, encoding a cellodextrin-cleaving beta-glucosidase and a xylodextrin degrading xylo-beta-glucosidase, respectively, were located directly adjacent to each other. The 5' region contained two additional genes, cglF and cglG, whose products exhibited similarity to integral membrane proteins of metabolite transport systems. The two beta-glucosidases, CglT and XglS, with deduced molecular masses of 52 and 81 kDa, belong to different families of glycosyl hydrolases. Both enzymes were overexpressed in Escherichia coli and could be detected after protein gel electrophoresis and activity staining. The enzyme CglT was purified by fast protein liquid chromatography and identified by N-terminal sequencing. The enzyme was thermostable at 60 degrees C for at least 24 h, and the temperature optimum was 75 degrees C. The ki for glucose inhibition was calculated to 200 mM. The enzyme released glucose from the nonreducing end of beta-1,4-cello oligomers as well as from various disaccharides. CglT was active on glucosides, galactosides and on fucosides, while XglS cleaved beta-glucosides and beta-xylosides as well. The cglT gene was also expressed in Bacillus subtilis, and the enzyme was mainly intracellular during exponential growth but was efficiently released into the supernatant after cultures entered the stationary phase. PMID- 9327556 TI - Comparison of phenanthrene and pyrene degradation by different wood-decaying fungi. AB - The degradation of phenanthrene and pyrene was investigated by using five different wood-decaying fungi. After 63 days of incubation in liquid culture, 13.8 and 4.3% of the [ring U-14C]phenantherene and 2.4 and 1.4% of the [4,5,9,10 14C]pyrene were mineralized by Trametes versicolor and Kuehneromyces mutabilis, respectively. No 14CO2 evolution was detected in either [14C]phenanthrene or [14C]pyrene liquid cultures of Flammulina velutipes, Laetiporus sulphureus, and Agrocybe aegerita. Cultivation in straw cultures demonstrated that, in addition to T. versicolor (15.5%) and K. mutabilis (5.0%), L. sulphureus (10.7%) and A. aegerita (3.7%) were also capable of mineralizing phenanthrene in a period of 63 days. Additionally, K. mutabilis (6.7%), L. sulphureus (4.3%), and A. aegerita (3.3%) mineralized [14C]pyrene in straw cultures. The highest mineralization of [14C] pyrene was detected in straw cultures of T. versicolor (34.1%), which suggested that mineralization of both compounds by fungi may be independent of the number of aromatic rings. Phenanthrene and pyrene metabolites were purified by high-performance liquid chromatography and identified by UV absorption, mass, and 1H nuclear magnetic resonance spectrometry. Fungi capable of mineralizing phenanthrene and pyrene in liquid culture produced enriched metabolites substituted in the K region (C-9,10 position of phenanthrene and C-4,5 position of pyrene), whereas all other fungi investigated produced metabolites substituted in the C-1,2, C-3,4, and C-9,10 positions of phenanthrene and the C-1 position of pyrene. PMID- 9327557 TI - Endosymbionts of ticks and their relationship to Wolbachia spp. and tick-borne pathogens of humans and animals. AB - The presence, internal distribution, and phylogenetic position of endosymbiotic bacteria from four species of specific-pathogen-free ticks were studied. These included the hard ticks Ixodes scapularis (the black-legged tick), Rhipicephalus sanguineus (the brown dog tick), and Haemaphysalis longicornis and the African soft tick Ornithodoros moubata. PCR assays for bacteria, using two sets of general primers for eubacterial 16S and 23S rRNA genes (rDNAs) and seven sets of specific primers for wolbachial, rickettsial, or Francisella genes, indicated that I. scapularis possessed symbiotic rickettsiae in the ovaries and that the other species harbored eubacteria in both the ovaries and Malpighian tubules. Phylogenetic analysis based on the sequence of 16S rDNA indicated that the symbiont of I. scapularis belonged to the alpha subgroup of proteobacteria and was closely related to the members of the genus Rickettsia. The other species had similar microorganisms in the ovaries and Malpighian tubules, which belonged to the gamma subgroup of proteobacteria, and formed a monophyletic group with the Q fever pathogen, Coxiella burnetii. O. moubata harbored another symbiont, which formed a monophyletic group with Francisella tularensis and Wolbachia persica, the latter a symbiont previously isolated from Malpighian tubules of the soft tick Argas (Persicargas) arboreus. Thus, the symbionts of these four tick species were not related to the Wolbachia species found in insects. The two symbionts that live in the Malpighian tubules, one closely related to C. burnetii and the other closely related to F. tularensis, appear to be of ancient origin and be widely distributed in ticks. PMID- 9327558 TI - Characterization of an endosymbiont infecting wood ticks, Dermacentor andersoni, as a member of the genus Francisella. AB - A microorganism (Dermacantor andersoni symbiont [DAS]) infecting Rocky Mountain wood ticks (D. andersoni) collected in the Bitterroot Mountains of western Montana was characterized as an endosymbiont belonging to the genus Francisella. Previously described as Wolbachia like, the organism's DNA was amplified from both naturally infected tick ovarial tissues and Vero cell cultures by PCR assay with primer sets derived from eubacterial 16S ribosomal DNA (rDNA) and Francisella membrane protein genes. The 16S rDNA gene sequence of the DAS was most similar (95.4%) to that of Francisella tularensis subsp. tularensis. Through a combination of Gimenez staining, PCR assay, and restriction fragment length polymorphism analysis, 102 of 108 female ticks collected from 1992 to 1996 were infected. Transovarial transmission to female progeny was 95.6%, but we found no evidence of horizontal transmission. PMID- 9327559 TI - Effect of fluorochromes on bacterial surface properties and interaction with granular media. AB - Simple, efficient, and safe tagging methods are desired in short-term microbial transport studies such as in the study of filtration systems for water and wastewater treatment. Suitability of selected fluorochromes as bacterial tagging agents in transport studies was evaluated on the basis of stability of stained cells and the effect of staining on bacterial surface characteristics and interaction with granular media. Surface properties were characterized by zeta potential and microbial adhesion to hydrocarbons. The effect of staining on interactions between bacteria and porous media was evaluated in terms of removal of bacteria in batch adsorption tests using sand coated with aluminum hydroxide to enhance adsorption. The DNA-specific fluorochrome 4',6-diamidino-2 phenylindole (DAPI) had generally negligible effects on bacterial surface properties and interaction with sand, as indicated in batch adsorption tests using pure cultures (Escherichia coli or Acinetobacter sp.) and wastewater bacteria. Cells stained with DAPI were stable for 48 h at 4 or 20 degrees C. Other nucleic acid fluorochromes tested had different but significant effects on bacterial cells and produced less stable fluorescence. Since transport through porous media is modulated by surface properties, it may be concluded based on these results that the choice of fluorochromes is critical in microbial transport studies. DAPI appeared to be a promising tagging agent. Time dependence of fluorescence of stained cells may limit the use of fluorochrome-tagged cells in long-term transport studies. PMID- 9327560 TI - Prevalence of and associated risk factors for shedding Cryptosporidium parvum oocysts and Giardia cysts within feral pig populations in California. AB - Populations of feral pigs (Sus scrofa) may serve as an environmental reservoir of Cryptosporidium parvum oocysts and Giardia sp. cysts for source water. We conducted a cross-sectional study to determine the prevalence of and associated demographic and environmental risk factors for the shedding of C. parvum oocysts and Giardia sp. cysts. Feral pigs were either live-trapped or dispatched from 10 populations located along the coastal mountains of western California, and fecal samples were obtained for immunofluorescence detection of C. parvum oocysts and Giardia sp. cysts. We found that 12 (5.4%) and 17 (7.6%) of 221 feral pigs were shedding C. parvum oocysts and Giardia sp. cysts, respectively. The pig's sex and body condition and the presence of cattle were not associated with the probability of the shedding of C. parvum oocysts. However, younger pigs (< or = 8 months) and pigs from high-density populations (> 2.0 feral pigs/km2) were significantly more likely to shed oocysts compared to older pigs (> 8 months) and pigs from low-density populations (< or = 1.9 feral pigs/km2). In contrast, none of these demographic and environmental variables were associated with the probability of the shedding of Giardia sp. cysts among feral pigs. These results suggest that given the propensity for feral pigs to focus their activity in riparian areas, feral pigs may serve as a source of protozoal contamination for surface water. PMID- 9327561 TI - Use of alkaline phosphatase as a reporter polypeptide to study the role of the subtilin leader segment and the SpaT transporter in the posttranslational modifications and secretion of subtilin in Bacillus subtilis 168. AB - The subtilin leader segment of presubtilin was fused to alkaline phosphatase (AP), which was used as a reporter polypeptide to study the role of the subtilin leader segment in posttranslational modifications during the conversion of presubtilin to subtilin and in the translocation of presubtilin from the cytoplasm of Bacillus subtilis 168 to the extracellular medium. It was observed that the subtilin leader segment could be utilized by a wild-type transporter, but secretion was enhanced if the subtilin transporter was available. The subtilin leader was not cleaved away from the AP component of the precursor until the precursor had been transported to the cell wall, and none of the AP was released into the medium until after cleavage had occurred. The role of SpaT, which is an ABC transporter that has been implicated in subtilin secretion, was explored by making a large in-frame deletion from the central region of SpaT and observing the effect on translocation of the AP reporter. Instead of having an effect on translocation, the deletion disrupted proteolytic cleavage of the subtilin leader segment and release of the AP reporter into the medium. The AP that was secreted by means of the subtilin leader segment had not undergone any posttranslational modifications, as assessed by amino acid composition analysis and enzymatic activity analysis. PMID- 9327562 TI - Single-site mutations in the conserved alternating-arginine region affect ionic channels formed by CryIAa, a Bacillus thuringiensis toxin. AB - The role of the third domain of CryIAa, a Bacillus thuringiensis insecticidal toxin, in toxin-induced membrane permeabilization in a receptor-free environment was investigated. Planar lipid bilayer experiments were conducted with the parental toxin and five proteins obtained by site-directed mutagenesis in block 4, an arginine-rich, highly conserved region of the protein. Four mutants were constructed by replacing the first arginine in position 21 by a lysine (R521K), a glutamine (R521Q), a histidine (R521H), or a glutamic acid (R521E). A fifth mutant was obtained by replacing the fourth arginine by a lysine (R527K). Like CryIAa, the mutants formed cation-selective channels. A limited but significant reduction in channel conductance was observed for all mutants except R521H. The effect was more dramatic for the voltage dependence of the channels formed by R521K and R521Q, which was reversed compared to that of the parental toxin. This study provides the first direct evidence of a functional role for domain III in membrane permeabilization. Our results suggest that residues of the positive arginine face of block 4 interact with domain I, the putative pore-forming region of CryIAa. PMID- 9327563 TI - Ribotyping for strain characterization of Clostridium perfringens isolates from food poisoning cases and outbreaks. AB - Ribotyping was used to characterize 34 Clostridium perfringens strains isolated from 10 food poisoning cases and outbreaks over a 7-year period. Twelve different ribopatterns were generated by EcoRI digestion. In eight food poisoning cases and outbreaks, all of the ribotypes of each food and stool isolate were found to be identical. Two C. perfringens isolates showed unique patterns. Ribotyping was found to be a useful tool for determining the genetic relationship of C. perfringens isolates in the context of foodborne poisoning cases. PMID- 9327564 TI - Size and complexity of the nuclear genome of Colletotrichum graminicola. AB - DNA reassociation was used to estimate GC content, size, and complexity of the nuclear genomes of Colletotrichum from maize and sorghum. Melting-temperature analysis indicated that the GC content of the maize pathotype DNA was 51% and that the GC content of the sorghum pathotype was 52%. DNA reassociation kinetics employing S1 nuclease digestion and an appropriately modified second-order equation indicated that the genome sizes of the maize and sorghum pathotypes were 4.8 x 10(7) bp, and 5.0 x 10(7) bp, respectively. Genomic reconstruction experiments based on Southern blot hybridization between a cloned single-copy gene, PYR1 (orotate phosphoribosyl transferase), and maize-pathotype DNA confirmed the size of the nuclear genome. The single-copy component of the genomes of both pathotypes was estimated at about 90%. For both pathotypes, ca. 7% of the genome represented repetitive DNA, and 2 to 3% was foldback DNA. PMID- 9327565 TI - Physiological basis for the high salt tolerance of Debaryomyces hansenii. AB - The effects of KCl, NaCl, and LiCl on the growth of Debaryomyces hansenii, usually considered a halotolerant yeast, and Saccharomyces cerevisiae were compared. KCl and NaCl had similar effects on D. hansenii, indicating that NaCl created only osmotic stress, while LiCl had a specific inhibitory effect, although relatively weaker than in S. cerevisiae. In media with low K+, Na+ was able to substitute for K+, restoring the specific growth rate and the final biomass of the culture. The intracellular concentration of Na+ reached values up to 800 mM, suggesting that metabolism is not affected by rather high concentrations of salt. The ability of D. hansenii to extrude Na+ and Li+ was similar to that described for S. cerevisiae, suggesting that this mechanism is not responsible for the increased halotolerance. Also, the kinetic parameters of Rb+ uptake in D. hansenii (Vmax, 4.2 nmol mg [dry weight]-1 min-1; K(m), 7.4 mM) indicate that the transport system was not more efficient than in S. cerevisiae. Sodium (50 mM) activated the transport of Rb+ by increasing the affinity for the substrate in D. hansenii, while the effect was opposite in S. cerevisiae. Lithium inhibited Rb+ uptake in D. hansenii. We propose that the metabolism of D. hansenii is less sensitive to intracellular Na+ than is that of S. cerevisiae, that Na+ substitutes for K+ when K+ is scarce, and that the transport of K+ is favored by the presence of Na+. In low K+ environments, D. hansenii behaved as a halophilic yeast. PMID- 9327566 TI - Gene sequence and expression of an analog of proliferating cell nuclear antigen (PCNA) in the alga Tetraselmis chui and detection of the encoded protein with anti-rat PCNA monoclonal antibody. AB - To identify a phytoplankton cell cycle marker detected by a monoclonal antibody against mammalian proliferating cell nuclear antigen (PCNA) (S. Lin, J. Chang, and E. J. Carpenter, J. Phycol. 30:449-456, 1994), a PCNA gene fragment was isolated by reverse transcription-PCR from the marine unicellular alga Tetraselmis chui Butcher (Prasinophyceae). The gene fragment was 616 bp in length and contained an open reading frame of 205 amino acids. The deduced amino acid sequence showed 80 and 88% similarity to human and rice PCNA, respectively. Southern hybridization indicated that the isolated gene fragment was part of the T. chui genome, with up to three copies in each haploid nucleus. Northern hybridization was used to detect a PCNA mRNA with a size of 1.2 kb from an exponentially growing algal culture. The T. chui gene fragment has been cloned into an expression vector, and a fusion protein was subsequently generated. Anti rat PCNA simultaneously recognized the PCNA fusion protein and a single 33-kDa band in T.chui total protein extract. Our results indicate that T. chui PCNA is highly similar to its mammalian counterpart and that anti-rat PCNA is a good tool for detecting phytoplankton PCNA in general. PMID- 9327567 TI - Impact of trichloroethylene and toluene on nitrogen cycling in soil. AB - The effects of trichloroethylene (TCE) and toluene on soil nitrogen-cycling activities were examined. Ammonium oxidation potential (AOP) was reduced after incubation with as little as 1 microgram of TCE ml-1, and the effects were generally greater when toluene was present and increased with longer exposure. Arginine ammonification potential and denitrification enzyme activity were constant regardless of TCE concentration or the presence of toluene, while nitrite oxidation potential (NOP) exhibited variable sensitivity. KCl-extractable ammonium levels increased dramatically after exposure to 30 and 60 micrograms of TCE ml-1 in the presence of toluene, whereas gamma-irradiated or sodium azide treated soil incubated with the same concentrations of TCE and toluene showed no increase. Alfalfa-amended soils showed similar decreases in AOP and increases in extractable ammonium during incubation with 60 micrograms of TCE ml-1 and 20 micrograms of toluene ml-1, although most probable number estimates of the ammonium oxidizer population showed no difference between exposed and unexposed soil. AOP and extractable ammonium returned slowly to control levels after 28 days of incubation in the presence of TCE and toluene. Activity assays to which various TCE and toluene concentrations were added indicated that AOP and NOP were relatively more sensitive to these compounds than was arginine ammonification potential. These results indicate that the soil microbial populations responsible for nitrogen cycling exhibit different sensitivities to TCE and toluene and that they may be more susceptible to adverse effects than previously thought. PMID- 9327568 TI - Multiple pathways for toluene degradation in Burkholderia sp. strain JS150. AB - Burkholderia (Pseudomonas) sp. strain JS150 uses multiple pathways for the metabolism of catechols that result from degradation of aromatic compounds. This suggests that the strain also uses multiple upstream pathways for the initial hydroxylation of aromatic substrates. Two distinct DNA fragments that allowed Pseudomonas aeruginosa PAO1c to grow with benzene as a sole carbon source were cloned from strain JS150. One of the recombinant plasmids containing the initial steps for the degradative pathway contained a 14-kb DNA insert and was designated pRO2016. We have previously shown that the DNA insert originated from a plasmid carried by strain JS150 and contained genes encoding a multicomponent toluene-2 monooxygenase (tbmABCDEF) as well as the cognate regulatory protein (tbmR) that controls expression of the 2-monooxygenase (G. R. Johnson and R. H. Olsen, Appl. Environ. Microbiol. 61:3336-3346, 1995). Subsequently, we have identified an additional region on this DNA fragment that encodes toluene-4-monooxygenase activity. The toluene-4-monooxygenase activity was also regulated by the tbmR gene product. A second DNA fragment that allowed P. aeruginosa to grow with benzene was obtained as a 20-kb insert on a recombinant plasmid designated pRO2015. The DNA insert contained genes encoding toluene-4-monooxygenase activity but no toluene-2-monooxygenase activity. The pRO2015 insert originated from the chromosome of strain JS150, unlike the region cloned in pRO2016. Southern blots and restriction map comparisons showed that the genes for the individual 4 monooxygenases were distinct from one another. Thus, strain JS150 has been shown to have at least three toluene/benzene monooxygenases to initiate toluene metabolism in addition to the toluene dioxygenase reported previously by others. PMID- 9327569 TI - Development and characterization of a whole-cell bioluminescent sensor for bioavailable middle-chain alkanes in contaminated groundwater samples. AB - A microbial whole-cell biosensor was developed, and its potential to measure water-dissolved concentrations of middle-chain-length alkanes and some related compounds by bioluminescence was characterized. The biosensor strain Escherichia coli DH5 alpha(pGEc74, pJAMA7) carried the regulatory gene alkS from Pseudomonas oleovorans and a transcriptional fusion of PalkB from the same strain with the promoterless luciferase luxAB genes from Vibrio harveyi on two separately introduced plasmids. In standardized assays, the biosensor cells were readily inducible with octane, a typical inducer of the alk system. Light emission after induction periods of more than 15 min correlated well with octane concentration. In well-defined aqueous samples, there was a linear relationship between light output and octane concentrations between 24 and 100 nM. The biosensor responded to middle-chain-length alkanes but not to alicyclic or aromatic compounds. In order to test its applicability for analyzing environmentally relevant samples, the biosensor was used to detect the bioavailable concentration of alkanes in heating oil-contaminated groundwater samples. By the extrapolation of calibrated light output data to low octane concentrations with a hyperbolic function, a total inducer concentration of about 3 nM in octane equivalents was estimated. The whole-cell biosensor tended to underestimate the alkane concentration in the groundwater samples by about 25%, possibly because of the presence of unknown inhibitors. This was corrected for by spiking the samples with a known amount of an octane standard. Biosensor measurements of alkane concentrations were further verified by comparing them with the results of chemical analyses. PMID- 9327570 TI - Hydroxyapatite adherence as a means to concentrate bacteria. AB - Adherence to hydroxyapatite (HA) was examined as a method to concentrate bacteria from foods. Using HA at a level of 10% and suspensions of an Escherichia coli strain containing 10(9), 10(6), and 10(3) cells per ml, kinetic studies revealed that maximum adherence was attained within 5 min for all cell concentrations and that comparable log reductions (1.0 to 1.5) of cells in suspension were seen regardless of initial cell concentration. Eleven species of spoilage and pathogenic bacteria were found to adhere to HA, with seven species adhering at proportions of greater than 95%. Fluorescent viability staining revealed that cells bound to HA remained viable. There was greater than 92% adherence of indigenous bacteria to HA from three of five 1:10 dilutions of ground beef, indicating promise for the use of HA for concentrating bacteria from meat and other food samples. PMID- 9327571 TI - Cadmium removal by a new strain of Pseudomonas aeruginosa in aerobic culture. AB - A fluorescent pseudomonad (strain CW-96-1) isolated from a deep-sea vent sample grew at 30 degrees C under aerobic conditions in an artificial seawater medium and tolerated cadmium concentrations up to 5 mM. After 140 h, strain CW-96-1 removed > 99% of the cadmium from solution. Energy dispersive microanalysis revealed that the cadmium was removed by precipitation on the cell wall; sulfide production was confirmed by growth on Kligler's agar. Based on 16S ribosomal DNA sequencing and fatty acid analysis, the microorganism is closely related to Pseudomonas aeruginosa. PMID- 9327573 TI - Survival of Escherichia coli cells exposed to iodoacetate and chlorodinitrobenzene is independent of the glutathione-gated K+ efflux systems KefB and KefC. AB - The KefB and KefC systems of Escherichia coli cells are activated by iodoacetate (IOA) and chlorodinitrobenzene (CDNB), leading to a rapid drop in the intracellular pH. However, survival of exposure to IOA or CDNB was found to be essentially independent of KefB and KefC activation. No correlation was found between the toxicity of the compound and its ability to elicit protective acidification via activation of KefB and KefC. PMID- 9327572 TI - Purification and characterization of a novel NADP-dependent branched-chain alcohol dehydrogenase from Saccharomyces cerevisiae. AB - An NADP-dependent branched-chain alcohol dehydrogenase was purified from Saccharomyces cerevisiae var. uvarum grown under anaerobic conditions. Its quaternary structure is monomeric, and it has a molecular mass of 37 kDa and a pI of 5.9. A possible role of the enzyme in flavor production during alcoholic fermentation is discussed. PMID- 9327574 TI - Effect of simulated microgravity and shear stress on microcin B17 production by Escherichia coli and on its excretion into the medium. AB - Production of the antibacterial polypeptide microcin B17 (MccB17) by Escherichia coli ZK650 was inhibited by simulated microgravity. The site of MccB17 accumulation was found to be different, depending on whether the organism was grown in shaking flasks or in rotating bioreactors designed to establish a simulated microgravity environment. In flasks, the accumulation was cellular, but in the reactors, virtually all the microcin was found in the medium. The change from a cellular site to an extracellular one was apparently not a function of gravity, since extracellular production occurred in these bioreactors, irrespective of whether they were operated in the simulated microgravity or normal gravity mode. More probably, excretion is due to the much lower degree of shear stress in the bioreactors. Addition of even a single glass bead to the 50 ml medium volume in the bioreactor created enough shear to change the site of MccB17 accumulation from the medium to the cells. PMID- 9327575 TI - Escherichia coli in settled-dust and air samples collected in residential environments in Mexico City. AB - Escherichia coli, an important indicator of the presence of fecal material, was isolated from indoor and outdoor environments in Mexico City. The heterogeneity of E. coli was represented by 89 serotypes, most of them coming from settled-dust indoor samples; 21% of them presented antibiotic multiresistance. The numbers of plasmids were higher among the antibiotic-resistant strains. The results of this study suggest that intestinal infections produced by environmental strains could be of more epidemiological impact than previously thought. PMID- 9327576 TI - Reduction of malachite green to leucomalachite green by intestinal bacteria. AB - Intestinal microfloras from human, rat, mouse, and monkey fecal samples and 14 pure cultures of anaerobic bacteria representative of those found in the human gastrointestinal tract metabolized the triphenylmethane dye malachite green to leucomalachite green. The reduction of malachite green to the leuco derivative suggests that intestinal microflora could play an important role in the metabolic activation of the triphenylmethane dye to a potential carcinogen. PMID- 9327577 TI - Phenotypic and genotypic biotyping of environmental strains of Vibrio cholerae non-O1 isolated in Italy. AB - The purpose of this study was to characterize strains of Vibrio cholerae non-O1 isolated in Italy from different sources by biochemical and serological assays, antibiotic susceptibility testing, and molecular biotyping. Serotyping and genomic analysis by pulsed-field gel electrophoresis proved to be useful in discriminating the isolates. The data obtained show a wide heterogeneity at the genomic level, and in keeping with this, the serogrouping classification provided evidence of a high variability of the investigated strains. In addition, none of the strains tested produced cholera-like toxins. PMID- 9327578 TI - Magnification of tributyl tin toxicity to oyster larvae by bioconcentration in biofilms of Shewanella colwelliana. AB - The toxic effects of dissolved versus bioconcentrated tributyl tin (TBT) on oyster larvae were compared. Water column TBT levels, which had no effect in solution, inhibited natural attachment and metamorphosis of oyster larvae on bottom surfaces due to bioconcentration by biofilms. This mechanism should be considered when evaluating heavy metal toxicity in the environment. PMID- 9327579 TI - Chemotaxis of Pseudomonas spp. to the polyaromatic hydrocarbon naphthalene. AB - Two naphthalene-degrading bacteria, Pseudomonas putida G7 and Pseudomonas sp. strain NCIB 9816-4, were chemotactically attracted to naphthalene in drop assays and modified capillary assays. Growth on naphthalene or salicylate induced the chemotactic response. P. putida G7 was also chemotactic to biphenyl; other polyaromatic hydrocarbons that were tested did not appear to be chemoattractants for either Pseudomonas strain. Strains that were cured of the naphthalene degradation plasmid were not attracted to naphthalene. PMID- 9327580 TI - Inhibition of beta-galactosidase biosynthesis in Escherichia coli by tetracycline residues in milk. AB - Low levels of tetracyclines found as residues in milk inhibited the biosynthesis of beta-galactosidase in Escherichia coli. To produce the same effect, other antibacterials had to occur in concentrations that were more than 10-fold higher. This relative selectivity was exploited for the development of a screening test for tetracyclines in milk based on a chemiluminometric assay of beta galactosidase. The method was validated with spiked samples of raw milk and applied to field samples contaminated with tetracyclines. PMID- 9327581 TI - Factors controlling acid tolerance of Listeria monocytogenes: effects of nisin and other ionophores. AB - The acid tolerance of a Listeria monocytogenes serotype 4b strain was studied by measuring its ability to survive at an acidic pH at 37 degrees C. The acid tolerance of L. monocytogenes was much lower than those of Escherichia coli O157:H7 and Shigella flexneri strains. This observation suggested a higher infective dose for L. monocytogenes than E. coli O157:H7 and Shigella. The susceptibility of L. monocytogenes to acidic pH was dependent upon growth medium pH and growth phase of the culture. Nisin and some other ionophores reduced the acid tolerance of both stationary-phase and log-phase cultures of L. monocytogenes. These studies indicated that nisin might be a useful candidate for controlling acid tolerance of L. monocytogenes. PMID- 9327582 TI - A simple filtration technique to detect enterohemorrhagic Escherichia coli O157:H7 and its toxins in beef by multiplex PCR. AB - Primers, specific for a unique base substitution in uidA of Escherichia coli O157:H7, were coupled with oligonucleotides for the shiga-like toxin I (SLT-I) and SLT-II genes in a multiplex PCR assay. A minimum of 10(2) CFU per PCR (10 microliters) was necessary to amplify E. coli O157:H7-specific bands by multiplex PCR. Food particles as well as various unknown metabolic by-products of bacteria inhibited the PCR, but a simple two-step filtration procedure eliminated this inhibition. To reliably generate PCR products, an E. coli inoculum of 10(3) CFU g of food slurry-1 in a nonspecific medium was required with 6 h of enrichment at 37 degrees C. However, when the food homogenate was incubated overnight, E. coli O157:H7 at an initial inoculum of even 1 CFU g-1 was detected. PMID- 9327583 TI - Detection of Mycobacterium ulcerans in environmental samples during an outbreak of ulcerative disease. AB - Mycobacterium ulcerans is an environmental bacterium which causes chronic skin ulcers. Despite significant epidemiological evidence to suggest that water is the source of infection, the organism has never been identified in the environment. Environmental water samples were collected from a small town in which an outbreak of 29 cases had occurred in a 3-year period. These were examined by mycobacterial culture and PCR amplification. Similar to previous studies, M. ulcerans was not cultured from the water samples. However, five samples were positive for M. ulcerans by PCR. These samples were collected from a swamp and a golf course irrigation system within the outbreak area. This is the first time that M. ulcerans has been demonstrated to be present in the environment and supports the postulated epidemiology of disease due to this organism. PMID- 9327584 TI - Structure of the gypsy moth vitellogenin gene. AB - Genomic clones containing the vitellogenin (Vg) gene from the gypsy moth were isolated from two genomic libraries and characterized. The nucleotide sequence of a 16,132 bp region of the gypsy moth genome was determined which included a 3,666 bp region upstream from the transcription initiation site and 499 bp region downstream from the transcribed region. Primer extension analysis was performed to identify the transcription initiation site. Gene sequence confirmed the sequence of VgmRNA recently reported [Hiremath and Lehtoma, J. Insect Biochem. Mol. Biol. (1997) 27:27-35] and indicated that the gypsy moth Vg gene contains seven exons interrupted by six introns. Sequence analysis of the promoter region revealed presence of several motifs associated with sex-specific and developmentally regulated genes in other systems. The nucleotide sequence comparison analyses showed that the gypsy moth Vg gene had considerably similarity with the Bombyx mori Vg gene but not with those from Anthonomous grandis and Aedes aegypti. PMID- 9327585 TI - Effect of demethylation on the chitinase inhibitory activity of allosamidin. AB - Removal of a methyl group of the allosamizoline moiety of allosamidin decreases the inhibitory effect on family 18 chitinases from three different species (a bacterium, Serratia marcescens, a crustacean, Artemia salina, and an insect cell line, Chironomus tentans). Loss of a second methyl group weakens enzyme inhibition further. This is in agreement with the highly conserved catalytic centre of these enzymes. PMID- 9327586 TI - Purification and characterization of a carboxylesterase associated with organophosphate resistance in the greenbug, Schizaphis graminum (Homoptera: Aphididae). AB - The Type II esterase associated with organophosphate resistance in the greenbug, Schizaphis graminum, was purified by column chromatography and preparative electrophoresis resulting in over 100-fold purification and approximately 11% recovery. The native enzyme appears to exist as a heterodimer with the subunits equal to 52 and 56 kDa. The mass of the native enzyme was estimated at 102 kDa by gel filtration chromatography and the isoelectric focusing point was 4.8. The enzyme was inhibited by both paraoxon and mercuric chloride, suggesting that it is a serine hydrolase, although it was not inhibited by carbamate insecticides or eserine. The enzyme was active toward both beta- and alpha-naphthol esters, although the length of the side chain (C-2 or C-4) also affected activity. The enzyme displayed no activity toward acetylthiocholine. N-Terminal amino acid sequence analysis of the enzyme subunits indicates that residues Val-4 and Gly-10 of the larger fragment were highly conserved among 11 other carboxylesterase sequences. Sequence data from the smaller fragment did not reveal any similarity with other esterase sequences. PMID- 9327587 TI - Filipino personality structure and the big five model: a lexical approach. AB - In lexically based studies, we derived Filipino personality dimensions and related them to the Big Five model. In Study 1, Filipino high-school and college students (N = 629) rated themselves on a near-comprehensive list of 861 Filipino (Tagalog) trait adjectives. In Study 2, Filipino high-school and college students (N = 1,531) rated 280 markers of dimensions identified in Study 1. Some students (n = 473) also completed the NEO Five-Factor Inventory. Seven comparable Filipino dimensions were identified in factor analyses in the two studies. We concluded that the dimensions we labeled Concern for Others (vs. Egotism), Conscientiousness, Gregariousness, and Intellect were quite similar to Big Five Agreeableness, Conscientiousness, Extraversion, and Intellect, respectively. The Filipino Self-Assurance dimension was most similar to Big Five Neuroticism. The Filipino Temperamentalness dimension was more complex in Big Five terms, overlapping Agreeableness, Conscientiousness, and Neuroticism. A final Filipino factor resembled a Negative Valence or Infrequency dimension. More than five factors had to be extracted to obtain Philippine dimensions resembling all of the Big Five. PMID- 9327588 TI - On energy, personality, and health: subjective vitality as a dynamic reflection of well-being. AB - In this article, we examine subjective vitality, a positive feeling of aliveness and energy, in six studies. Subjective vitality is hypothesized to reflect organismic well-being and thus should covary with both psychological and somatic factors that impact the energy available to the self. Associations are shown between subjective vitality and several indexes of psychological well-being; somatic factors such as physical symptoms and perceived body functioning; and basic personality traits and affective dispositions. Subsequently, vitality is shown to be lower in people with chronic pain compared to matched controls, especially those who perceive their pain to be disabling or frightening. Subjective vitality is further associated with self-motivation and maintained weight loss among patients treated for obesity. Finally, subjective vitality is assessed in a diary study for its covariation with physical symptoms. Discussion focuses on the phenomenological salience of personal energy and its relations to physical and psychological well-being. PMID- 9327589 TI - Predicting self-esteem, well-being, and distress in a cohort of gay men: the importance of cultural stigma, personal visibility, community networks, and positive identity. AB - Homosexual and bisexual men (N = 825) enrolled in the Multicenter AIDS Cohort Study in Chicago completed a 90-minute self-administered questionnaire that included the Rosenberg Self-Esteem Scale, a Well-Being Index, and the Hopkins Symptom Checklist. Participants indicated their experiences with gay stigma, their visibility as gay men, their involvement in the gay community, and their commitment to a positive gay identity. Data from this predominantly white, young, educated, and middle-class cohort are consistent with a structural model in which cultural stigma is negatively associated with positive self-perceptions. This within-group result contrasts sharply with between-group results that indicate our gay cohort was neither particularly low in global self-esteem nor high in psychological distress when compared to nonstigmatized samples. PMID- 9327590 TI - Personality, social networks, and perceived social support among alcoholics: a structural equation analysis. AB - In this study we tested relations among personality characteristics, social network properties, and perceived social support both concurrently and prospectively. A sample of 294 men in treatment at a Department of Veterans Affairs Alcohol Treatment Unit was assessed during treatment and 3 months after discharge. Results of the cross-sectional structural equation analyses indicated that the personality characteristics of extraversion and neuroticism were related to both social network properties and perceived social support. Characteristics of the alcoholic's social network were also related to perceived availability of support. Longitudinal analyses of perceived social support after treatment indicated that two social network properties (size of the network and the proportion of confidants) were predictive net of initial levels of social support. Extraversion and neuroticism were found to be indirectly related to perceived social support at Time 2 through their effects on social network properties and perceived social support during treatment. Implications of these findings for models of the nature and determinants of perceived social support are discussed. PMID- 9327591 TI - Ectopic expression of a tobacco homeobox gene, NTH15, dramatically alters leaf morphology and hormone levels in transgenic tobacco. AB - The shoot apical meristem functions to generate the lateral organs of a plant throughout the vegetative and reproductive phases. Homeobox genes play key roles in controlling such developmental programs, but their modes of action have not been well defined. Here we describe isolation and biological functions of a novel tobacco homeobox gene, designated NTH15 (Nicotiana tabacum homeobox 15), from a tobacco shoot apex cDNA library. NTH15 encodes a polypeptide of 342 amino acids, its homeodomain is very similar to the class 1 KNOTTED-type homeodomains. NTH15 mRNA is mainly localized in corpus cells in the tobacco shoot apical meristem, but not in tunica layers nor in differentiated lateral organs. The NTH15 cDNA was fused to the cauliflower mosaic virus 35S promoter and used to generate transgenic tobacco plants. Almost all transgenic tobacco plants showed abnormal leaf and/or flower morphology, and were categorized into three groups depending on severity of the leaf phenotype. In transgenic leaves, drastic decrease of GA1 and increase of cytokinin were observed, while the levels of other phytohormones were only slightly changed. Taken together, our results suggest NTH15 is involved in tobacco morphogenesis and abnormal leaf morphology in transgenic plants results from altered hormone levels. PMID- 9327592 TI - Differential expression of two genes for sucrose-phosphate synthase in sugarcane: molecular cloning of the cDNAs and comparative analysis of gene expression. AB - Two cDNA clones, pSoSPS1 and pSoSPS2, encoding sucrose-phosphate synthase (SPS) of sugarcane were isolated from a leaf cDNA library. Northern analysis revealed the transcript of SoSPS1 to be predominant in leaves, but that of SoSPS2 to be distributed not only in leaves but also in roots at a similar level. The transcript of SoSPS1 was markedly accumulated during greening of etiolated leaves, whereas that of SoSPS2 was constitutively expressed. These findings indicate that SPS in sugarcane is encoded by multiple genes, which show organ specificity and are differentially regulated in response to light. PMID- 9327593 TI - Expression of a gene for a protein similar to HIV-1 Tat binding protein 1 (TBP1) in floral organs of Brassica rapa. AB - A cDNA for a protein similar to human immunodeficiency virus Tat binding protein was isolated from an anther cDNA library of Brassica rapa. RNA in situ analysis in flower buds showed that the gene for this cDNA was specifically expressed in the tapetum and middle layer of anthers and pollen. PMID- 9327594 TI - The gene encoding flavanone 3-hydroxylase is expressed normally in the pale yellow flowers of the Japanese morning glory carrying the speckled mutation which produce neither flavonol nor anthocyanin but accumulate chalcone, aurone and flavanone. AB - The Japanese morning glory carrying the recessive mutable speckled allele with the dominant speckled-activator bears colorless flowers with fine and round colored spots distributed over the corolla whereas the plant without the speckled activator produces pale yellow flowers. Previous chemical analysis has indicated that a mutation in the gene for flavanone 3-hydroxylase (F3H) is a likely candidate for the speckled allele. However, the F3H mRNA without sequence alteration accumulates normally in the pale yellow flowers, indicating that the speckled allele is neither the F3H gene nor a regulatory gene acting on the F3H gene expression. PMID- 9327596 TI - Visualization of flumes from pharmaceutical delivery devices. AB - For the treatment of some diseases, drugs are delivered suspended in sprays that are administered either intranasally or by inhalation. The characteristics of these sprays or flumes have an influence on the effectiveness of the drug and some of these characteristics are best understood if the emerging flumes can be visualized. At GlaxoWellcome we have developed a multi-exposure photographic system whereby individual flumes can be recorded at pre-set intervals over their entire time period, typically 250 ms. It may also be desirable to visualize the flume within the nasal passage and this may be achieved by the use of a glass model nose. In this paper I will summarize pre-existing photographic methods and describe in detail the new technique. PMID- 9327597 TI - Photographing the cleft palate. PMID- 9327595 TI - An unusual mitochondrial atp9-rpl16 cotranscript found in the maternal distorted leaf mutant of Arabidopsis thaliana: implication of GUG as an initiation codon in plant mitochondria. AB - Properties of an unusual atp9-rpl16 cotranscript preferentially found in the maternal distorted leaf mutant of Arabidopsis thaliana, which had arisen from a genetic cross between chloroplast mutator and wild-type plants, were examined. Analysis of RNA editing of this cotranscript showed that one editing event in the rpl16 coding region created a UGA stop codon. This raises a possibility that a downstream GUG codon can serve as an initiation codon for rpl16. PMID- 9327598 TI - Medicine and television. AB - 1997 is the 50th anniversary of the first use of television for medical teaching and the 40th anniversary in Britain of an experimental 2-hour demonstration of pre-clinical teaching by colour television, of the use of large screen television for medical conferences, and of the presentation on national television of programmes aimed at modifying the public attitude to mental illness and to medicine. PMID- 9327599 TI - Ganglion. PMID- 9327600 TI - Communication in surgical illustration. AB - The surgeon is a unique user of medical illustration. With both hands busy, necessarily occupying the space with the best view of what's going on, muffled by a mask and hemmed in by operating theatre personnel, the surgeon is not well situated for teaching. On the other hand, teaching is precisely what medical illustrators do, through their drawings and diagrams: the profession has no other raison d'etre. Communication is essential, from surgeon to artist and from artist to student. Two unexpected obstacles are here discussed (only partly tongue-in cheek). PMID- 9327601 TI - Frank Netter (1906-1991). PMID- 9327602 TI - Ventricular defibrillating threshold: strength-duration and percent-success curves. AB - The term defibrillation threshold is usually understood to mean the shock intensity just enough to defibrillate a specified cardiac chamber (atria or ventricles). With the advent of so many different types of defibrillator, it is important to be able to specify the defibrillation threshold, which has frequently been described by the classical strength-duration curve. Another method of representing defibrillation plots the percent-successful defibrillation against shock-strength area. The mechanism of defibrillation is discussed, and the concepts of the strength-duration curve and percent-success against shock strength curves are compared. Because defibrillation is associated with a time varying spectrum of cellular excitability, a given shock strength will not always achieve defibrillation, and this produces the sigmoid shape for the curve that relates percent-successful defibrillation to shock strength. Therefore it is important to recognise two concepts: first, there is a family of strength duration curves for defibrillation, each curve representing a given percent successful defibrillation, and, secondly, there is a family of percent-success against shock-strength curves, one for each pulse duration. Canine ventricular defibrillation data are used to bring these two concepts together. Most importantly, the concepts adduced in the paper apply to transventricular, intracardiac and transchest defibrillation; the only difference in these applications is a scale factor that represents electrode location with respect to the heart. PMID- 9327603 TI - Time-frequency analysis of the first heart sound. Part 1: Simulation and analysis. AB - The authors propose a simulated first heart sound (S1) signal that can be used as a reference signal to evaluate the accuracy of time-frequency representation techniques for studying multicomponent signals. The composition of this simulated S1 is based on the hypothesis that an S1 recorded on the thorax over the apical area of the heart is composed of constant frequency vibrations from the mitral valve and a frequency modulated vibration from the myocardium. Essentially, the simulated S1 consists of a valvular component and a myocardial component. The valvular component is modelled as two exponentially decaying sinusoids of 50 Hz and 150 Hz and the myocardial component is modelled by a frequency modulated wave between 20 Hz and 100 Hz. The study shows that the simulated S1 has temporal and spectral characteristics similar to S1 recorded in humans and dogs. It also shows that the spectrogram cannot resolve the three components of the simulated S1. It is concluded that it is necessary to search for a better time-frequency representation technique for studying the time-frequency distribution of multicomponent signals such as the simulated S1. PMID- 9327604 TI - Time-frequency analysis of the first heart sound. Part 2: An appropriate time frequency representation technique. AB - A simulated first heart sound (S1) signal is used to determine the best technique for analysing physiological S1 from the following five time-frequency representations (TFR): the spectrogram, time-varying autoregressive modelling, binomial reduced interference distribution, Bessel distribution and cone-kernel distribution (CKD). To provide information on the time and frequency resolutions of each TFR technique, the instantaneous frequency and the -3 dB bandwidth as functions of time were computed for each simulated component of the S1. The performance index for selecting the best technique was based on the relative error and the correlation coefficient of the instantaneous frequency function between the theoretical distribution and the computed TFR. This index served to select the best technique. The sensitivity of each technique to noise and to small variations of the signal parameters was also evaluated. The results of the comparative study show that, although important limitations were found for all five TFRs tested, the CKD appears to be the best technique for the time-frequency analysis of multicomponent signals such as the simulated S1. PMID- 9327605 TI - New technique for time series analysis combining the maximum entropy method and non-linear least squares method: its value in heart rate variability analysis. AB - A new technique for time series analysis, which is a combination of the maximum entropy method (MEM) for spectral analysis and the non-linear least squares method (LSM) for fitting analysis, is described. In this technique, the MEM power spectral density (MEMPSD) is calculated using a very large lag that could diminish the lag dependence of dominant periods estimated by the MEM analysis. The validity of this large lag is confirmed by the LSM, given that the ten dominant MEM periods are known quantities. To validate the MEM plus LSM technique, it is compared with autoregressive (AR) modelling, by analysing heart rate variability under pharmacological interventions (phenylephrine and trinitroglycerine), using 16 young males. The results indicate that the MEMPSD, when compared with the ARPSD, has numerous periods that could reproduce the original time series much more accurately, as revealed by the LSM analysis. However, both the low- and high-frequency powers with MEMPSD and ARPSDs shift in the expected directions in accordance with the pharmacological effects on the cardiovascular system. The implications of these results are discussed from the theoretical and practical standpoints of the MEM plus LSM technique, compared with AR modelling. PMID- 9327607 TI - Fluctuations in the cerebral oxygenation state during the resting period in functional mapping studies of the human brain. AB - Functional mapping studies using near-infrared spectroscopy have detected for the first time the existence of significant fluctuations in the concentration of cerebral oxygenated [oxy-Hb], and deoxygenated haemoglobin, [deoxy-Hb], during the resting period. The fluctuations are not related to alterations in the systemic circulatory system. The temporal pattern varies with each brain region. In some instances, the degrees of change in [oxy-Hb] and [deoxy-Hb] caused by highly integrated tasks are within this resting variation range. Thus, taking account of these fluctuations is essential to the interpretation of distribution patterns of cerebral activity. PMID- 9327606 TI - Non-invasive determination of instantaneous heart rate in developing avian embryos by means of acoustocardiogram. AB - Previous noninvasive studies of the mean heart rate of embryonic birds have prompted an investigation into the instantaneous heart rate (IHR), which may be informative in developmental studies of cardiac rhythm. Using the acoustocardiogram (ACG), a noninvasive, long-term measuring system for embryonic IHR is developed, and the IHR in chickens during the last half of embryonic development is determined. The system, which uses a micro-computer, samples the ACG at a frequency of 50 Hz, restores the ACG wave by sinc function and calculates the IHR with an error in accuracy of less than 1 beat min-1. It was found that characteristic, transient bradycardia begins to appear late in the second week of incubation, and, with the additional development of transient tachycardia, the embryonic cardiac rhythm becomes more arrhythmic towards hatching. Simultaneous measurements of IHR with somatic movements showed no relationship between arrhythmia and embryonic activities. This system is useful, providing new evidence on long-term IHR developmental patterns. PMID- 9327608 TI - Measurement of venous oxyhaemoglobin saturation in the adult human forearm by near infrared spectroscopy with venous occlusion. AB - Measurement of the oxygenation of the peripheral tissues provides useful information about tissue perfusion. A method is described for the measurement of peripheral venous oxyhaemaglobin saturation (SvO2) in the adult forearm by a noninvasive technique, near infrared spectroscopy (NIRS) with venous occlusion. A series of studies is performed on healthy adults to compare measurements of forearm SvO2 made by NIRS with measurements of superficial venous SvO2 made by co oximetry, and to study the effect of different optode spacings. There is a significant correlation between forearm SvO2 measured by NIRS and SvO2 of superficial venous blood measured by co-oximetry (n = 19, r = 0.7, p < 0.0001). Higher values for SvO2 were obtained using a 2.5 cm spacing than with a 4 cm spacing (mean difference = 4.1% (95% CI 1.4%-6.8%) n = 16). This difference is likely to have been due to a more superficial volume of tissue being studied with the closer optode spacing. Peripheral SvO2 can be measured non-invasively using NIRS with venous occlusion. It may prove to be a useful method to study circulatory disturbances. PMID- 9327609 TI - Removal of catheter distortion in multiple indicator dilution studies: a deconvolution-based method and case studies on glucose blood-tissue exchange. AB - The study of blood-tissue exchange by the multiple indicator dilution technique often needs frequent sampling in the blood of the indicator dilution curves (IDC). Usually, this requires the use of a catheter supported by a pump. This causes a distortion in the IDC, which must be removed for proper interpretation of the data. A deconvolution-based methodology to remove IDC distortion is presented. First, the catheter impulse response is modelled by means of data obtained from a suitable experiment. Then the reconstruction of the blood IDC is tackled by a new nonparametric deconvolution algorithm, which provides (quasi) time-continuous signals and exploits statistically based criteria for the choice of the regularisation parameter. The methodology is applied to the removal of catheter distortion in studies of glucose blood-tissue exchange in the human forearm and myocardium. PMID- 9327611 TI - Impedance of a goat eye lens. AB - The complex electrical impedance of a goat eye lens is studied in the frequency range 10 mHz-10 Hz at room temperature, using a computer-controlled AC impedance system. AC impedance software (model 368, version 2.2) is employed to determine the total impedance and capacitance of the eye lens at various frequencies. A Cole-Cole plot of the eye lens material is drawn between the real component of impedance Z' and the imaginary component Z" for each excitation frequency that shows a perfect arc of a-semicircle, with its centre lying below the abscissa at an angle of 35 degrees. The half-angle phi between R(0) and R infinity is found to be 55 degrees, which mathematically demonstrates the selective permeability of the eye lens. Using graphical analysis of the Cole-Cole plot, characteristic frequency fc and distribution factor alpha are observed to be 1 Hz and 0.77, respectively. At characteristic frequency, capacitance and total impedance are found to be 1.14 microF and 9.08 k omega. The effect of electrode polarisation on capacitance is corrected, based on Fricke's power function. The observed electrical parameters are then used to explain the multiple current path through various tissue compartments. Further, an attempt is made to explain the results on the basis of a possible dipolar model. PMID- 9327610 TI - Estimation of habituation and signal-to-noise ratio of cortical evoked potentials to oesophageal electrical and mechanical stimulation. AB - Electrical and mechanical stimulation of the oesophagus has been recently proposed to examine the physiological effects of autonomic stimulation in humans. Cortical evoked potentials (EPs) to oesophageal stimulation provide an assessment of afferent fibres and central processing. However, habituation takes place during averaging of cortical EPs and reduces the signal-to-noise ratio (SNR) as the number of stimuli increases. The SNR of cortical EPs to oesophageal stimulation is computed for 15 normal subjects. Habituation is characterised by the Euclidean distance between the EEG response to single stimuli and the averaged EP, to serve as an objective measure of similarity between the averaged EP and the single-stimulus EEG. With electrical stimulation, the SNR is highest (0.41 +/- 0.21) for 1-12 stimuli and then significantly decreases to 0.2 +/- 0.08 for 13-24 stimuli (p < 0.001). With balloon distension (BD), the SNR is highest (0.22 +/- 0.16) for 1-12 stimuli and lowest (0.12 +/- 0.14) for 13-24 stimuli, but these SNRs are not significantly different from each other. Both electrical and mechanical stimulation of the oesophagus produce rapidly adapting EPs. The SNR of the EPs is higher with electrical stimulation than with BD. The EPs response to BD has a higher variability and is more noisy. Consequently, these results suggest that the overall cortical EP response to electrical stimulation of the oesophagus is more reproducible than that due to balloon distension. PMID- 9327612 TI - Spike detection in biomedical signals using midprediction filter. AB - Spikes such as QRS complex in ECG, epileptic seizures in EEG, fine crackles in vesicular sound and glottal closure instants in voiced sound are of diagnostic importance. Various methods of spike detection use the amplitude and frequency characteristics of the spikes. Because of the high frequency content, the spikes appear in the error signal when a linear prediction filtering scheme is used. The authors use the method of midprediction filtering for the detection of the spikes. In this method, the present sample is predicted as a weighted average of p recent past and p immediate future samples. The symmetrical nature of midprediction causes the spikes to appear in the error signal with their original basewidths. This can help in improving the reliability of spike detection, as both the amplitude and the duration of the spike can be considered as decision making parameters. It is observed that the high frequency gain of the midprediction filter is higher compared to the high frequency gain of the LPC or endprediction filter. As a result, this method works better than linear prediction for the detection of spikes. PMID- 9327613 TI - Acoustical recognition of laryngeal pathology using the fundamental frequency and the first three formants of vowels. AB - The recognition of laryngeal pathology by analysis of the voice is investigated. The fundamental frequency and the first three formants are considered. The recognition strategy is based on comparison with normal ranges calculated over 200 ordinary voices, grouped in ten age classes ranging from 20 to 70 years, for males and females. 220 test voices are studied divided into four groups: normal voices, functional dysphonia, nodules and recurrent nerve palsy. Each subject is marked according to his/her normal range. Parameters (or items) are calculated on the Interactive Laboratory System workstation. The vocalic material is composed of 11 vowels taken from a sentence. Results are given in terms of the number of values out of the normal ranges. Statistical analysis considers both parameter ability and error rates in pathology recognition. Pathology recognition shows the following error percentages: 23% for dysphonia, 14% for nodules and 33% for recurrent nerve palsy. Parameters do not show the same efficiency for voice pathology characterisation. Formants appear to be better than the fundamental frequency. PMID- 9327614 TI - In vitro 3D reconstruction of long bones using B-scan image processing. AB - An echographic image processing method has been developed, and validated by in vitro experiments, for the 3D reconstruction of the long bones of the newborn. The reconstruction of successive parallel cross-sections is obtained by a 2D reconstruction technique using radial B-scan image processing. The automatic segmentation of all the calculated images allows the extraction of the external contours of the skeleton. After structuring the explored volume using a contour association method, a contour interpolation step is required to solve the anisotropy problem, to obtain a 3D representation with cubic voxel lists. The results are encouraging, and a new mechanical part prototype of the acquisition system is under test for in vivo experiments. The main originality of the paper lies in the combination of different steps to obtain a practical solution to a clinical problem. PMID- 9327615 TI - Computerised multiparametric analysis from images of blood flow through frog mesenteric arterial bifurcation. AB - A computerised multiparametric procedure is developed to analyse the images of blood flow through various locations of the mesenteric arterial bifurcation of frog. The data are recorded by a video microscopic system and, after digitisation and pre-processing, are analysed by an IBM PC/AT based image processing system to obtain erythrocyte and velocity distribution profiles by axial tomographic and image velocimetry techniques, respectively. The vessel radius, haematocrit, blood flow through main and branch arteries and flow separation zones are determined from the data by various analytical procedures. In contrast to the earlier techniques the data are obtained from the same location of the vessel and thus the variability in flow parameters is minimised. PMID- 9327616 TI - Effect of ankle-foot orthosis on active ankle moment in patients with hemiparesis. AB - The paper investigates the effect of dorsi/plantar rigidity and the initial angle of ankle-foot orthoses (AFOs) on the moment generated by ankle musculature (referred to as active ankle moment) during gait in patients with hemiparesis. In the early stance phase, the active ankle moment in the direction of dorsiflexion is negligible, and AFOs play an important role in supplementing weak dorsiflexion. In mid to late stance, the moment generated by AFOs is very small compared with the active ankle moment in the direction of plantarflexion. AFOs therefore play only a limited role in assisting plantar flexors during this period. The active ankle moment in the direction of plantarflexion varies significantly with changes in the rigidity and initial angle of AFOs in 11 out of 20 subjects. The implication of this finding is discussed in relation to the need for dynamic matching of AFOs in individual patients with hemiparesis. PMID- 9327617 TI - Technology assessment using the association between outcome measures and patterns of illness severity. AB - The inclusion of a patient's illness experience as outcome in the assessment of health care technology has revealed methodological limitations such as the interpretation of multi-attribute scores and lack of knowledge about the association between illness and disease information. In an attempt to overcome these limitations, a cross-sectional study is performed to search for patterns of illness severity and investigate the association between illness measures and between illness patterns and disease factors. A sample of 211 patients with ulcerative colitis is studied using the sickness impact profile (SIP) and the rating form for inflammatory bowel disease patient concerns (RFIPC) as illness measures. SIP and RFIPC scores show low association, suggesting that they provide complementary information about the patient's illness status. Cluster analysis is performed using the two measures of illness separately to identify groups of patients with different degrees of severity of illness (clusters). The cluster description covers illness, disease and social and demographic variables. The RFIPC clusters show a general pattern of ascendant rank scores for the RFIPC items. SIP clusters differ, not only in the level of severity, but also in specific types of disability. The patients in the clusters with the highest degree of disability (reflected by SIP) show a non-linear relationship with patients' concerns (reflected by RFIPC) and disease factors. PMID- 9327618 TI - Clinical assessment of Hokkaido University PACS. AB - The paper describes the hardware and software used in the Hokkaido University picture archiving and communication system (HUPACS), which has been in use for the past two years. An evaluation of the system is also included. Fuji computed radiography completely replaces conventional X-ray radiographs, and 10:1 data compression is routinely employed. The PACS LAN is connected to the hospital information system through the main-frame computer and it is linked, not only to the radiology department, but also to the outpatient clinics. Image data have been steadily stored in the optical disc library, which contains more than 200,000 images. The original image can be accessed within 1 min. Image quality on a cathode ray tube (CRT) is clinically acceptable. Nevertheless, the transfer time of a newly made image is prolonged in some cases, during periods of heavy usage of the system. During scheduled radiology conferences, image reading tends to take more time on a CRT than on film. An improvement in the ease of operation of the workstations is necessary for full acceptance of CRT diagnoses. Generally speaking, however, HUPACS is working well and is appreciated by most of the clinicians. PMID- 9327619 TI - Design of a summary screen for an ICU patient data management system. AB - The paper describes a used-centred design for the summary screen of a computerised ICU patient data management system (PDMS). The screen also forms the resting state display, or default screen, and provides the principal navigation tool to other functionality within the system. The design process identified the most frequent potential users of this screen to be the nurses. Their tasks and the information resources required to perform them were analysed. The analysis identified that the nurses' main task of planning and implementing patient care required an awareness of a set of physiological parameters which provided an overview of the patient's general condition. Novel formats are proposed for displaying the trends in physiological parameters and these have been incorporated into a proposed screen design. These display formats have been evaluated by ICU nurses; they were adjudged to be clear, relevant, easy to learn and simple to use. Nurses considered the content of the screen, and the display formats used, to be suitable for maintaining an awareness of a patient's state during routine patient management. PMID- 9327620 TI - Biopotential amplifier for simultaneous operation with biomagnetic instruments. AB - A multichannel biopotential amplifier for simultaneous use with biomagnetic measurements in a magnetically shielded room is designed and evaluated. Particular care is taken to make the amplifier electromagnetically compatible with the biomagnetic instruments over the whole frequency spectrum, from DC to RF. The electromagnetically quiet environment allows the use of high electrode impedances; the preamplifier has been designed accordingly. Special care is taken to analyse the coupling mechanisms of mains interference to the amplifier. Over 170 simultaneous electric and magnetic recordings have been performed using the system in a hospital environment. PMID- 9327621 TI - Management of dental trauma: development of a 2D data acquisition system to evaluate passivity of dental splints. AB - Management of dental trauma in children sometimes requires the use of dental splints, which can be constructed from orthodontic materials. Past studies of the mechanical properties of dental splints have only been interested in their flexibility and not passivity. The purpose of this study is to determine the passivity of splints constructed from orthodontic materials. A specific data acquisition system, with a 2D transducer, is developed for evaluation of its neutrality. The transducer detects the displacements of the splinted tooth generated by the splint. The splints are constructed with 0.406 mm round, straight or Arch Blank preformed stainless-steel wires, with 0.559 mm standard edgewise brackets and with 0.254 mm stainless-steel or 3.05 mm elastomeric ligatures. Results show that mean output voltages generated by the splints range from 1.13 V to 2.48 V. The best control of passivity is obtained with the splints constructed with a preformed archwire and with elastomeric ligatures (p < 0.05), and the worst control is obtained with those constructed with a straight wire and with stainless-steel ligatures (p < 0.05). PMID- 9327622 TI - Information in full and reduced data sets of electrical impedance spectra from various skin conditions, compared using a holographic neural network. AB - Information relating to patho-physiological events in living tissues, including skin, should be found in the beta-dispersion frequency range of the electrical impedance. For intact skin, the alpha-dispersion is strongly influenced by the condition of the stratum corneum, particularly its hydration. For intact skin, these dispersions are not well separated: in a Nyquist plot, most of the relatively small beta-dispersion will be hidden behind the upper-frequency end of the alpha-dispersion arc. In the study, information obtained with a set of four indices, claimed to extract most of the information in the pertinent frequency interval, is compared with the full information measured at 31 frequencies, between 1 kHz and 1 MHz, for irritant and allergic contact reactions and nodular basal cell carcinomas, using a holographic neural network that appears to be useful for model-independent evaluation of the consequences of data-reduction procedures. Cole parameters should be avoided in the beta-range for intact skin. The indices are well supported, as long as differences from a reference site are used, and it seems that they can serve as the basis for differential diagnostics even as absolute values, although more information may be extracted from the complete spectrum. PMID- 9327623 TI - Optimisation algorithms for ECG data compression. AB - The use of exact optimisation algorithms for compressing digital electrocardiograms (ECGs) is demonstrated. As opposed to traditional time-domain methods, which use heuristics to select a small subset of representative signal samples, the problem of selecting the subset is formulated in rigorous mathematical terms. This approach makes it possible to derive algorithms guaranteeing the smallest possible reconstruction error when a bounded selection of signal samples is interpolated. The proposed model resembles well-known network models and is solved by a cubic dynamic programming algorithm. When applied to standard test problems, the algorithm produces a compressed representation for which the distortion is about one-half of that obtained by traditional time-domain compression techniques at reasonable compression ratios. This illustrates that, in terms of the accuracy of decoded signals, existing time domain heuristics for ECG compression may be far from what is theoretically achievable. The paper is an attempt to bridge this gap. PMID- 9327624 TI - Optical assessment of recovery of tissue blood supply after removal of externally applied pressure. AB - The authors use photoelectric plethysmography to determine the external occlusion pressure for blood vessels in human tissue in vivo. Three wavelengths are employed; 950 nm (infra-red), 640 nm (red) and 583 nm (yellow). Each probe is applied in turn to one finger of each subject. Pressure is applied, using a neonatal blood pressure cuff, to the finger via the probe. This pressure is increased linearly to 20 kPa (150 mmHg) over 15 s and then decreased linearly to zero over 15 s. The pressure at which perfusion returns is obtained for four repeat measurements at each wavelength. The mean (+/-standard deviation) occlusion pressures for all 13 subjects investigated are 7.1 (+/-1.9) kPa for infra-red, 6.3 (+/-1.7) kPa for red and 5.8 (+/-1.8) kPa for yellow. The pressure is 0.79 (+/-0.83) kPa lower for red compared with infra-red (P < 0.01), 0.54 (+/ 0.60) kPa lower for yellow compared with red (P < 0.002) and 1.3 (+/-1.0) kPa lower for yellow compared with infra-red (P < 0.005). The reduced penetration of shorter optical wavelengths can be used to detect the lower occlusion pressures of the smaller blood vessels nearer the skin surface. PMID- 9327625 TI - Hopfield network applied to blood vessel detection in angiograms. AB - A neural network classifier for detecting vascular structures in angiograms is developed. The classifier consists of a Hopfield network applied to a square window in which the centre pixel is classified from binary information within the window. Tests are performed using a binary test image corrupted by inverting a percentage of the image pixels. The resulting noisy images simulate the output of a detector using a simple threshold derived from local image statistics. The factors affecting the size of window and the choice of stored patterns are discussed. The results are compared with those obtained from a multi-layer perceptron using a similar approach. The Hopfield network is found to be effective at rejecting the high levels of noise that would result from low contrast source imagery. Another important feature is that the processed image retains an accurate representation of blood vessel diameter. PMID- 9327626 TI - Ballistocardiogram of avian eggs determined by an electromagnetic induction coil. AB - As an avian embryo grows within an eggshell, the whole egg is moved by embryonic activity and also by the embryonic heartbeat. A technical interest in detecting minute biological movements has prompted the development of techniques and systems to measure the cardiogenic ballistic movement of the egg or ballistocardiogram (BCG). In this context, there is interest in using an electromagnetic induction coil (solenoid) as another simple sensor to measure the BCG and examining its possibility for BCG measurement. A small permanent magnet is attached tightly to the surface of an incubated egg, and then the egg with the magnet is placed in a solenoid. Preliminary model analysis is made to design a setup of the egg, magnet and solenoid coupling system. Then, simultaneous measurement with a laser displacement measuring system, developed previously, is made for chicken eggs, indicating that the solenoid detects the minute cardiogenic ballistic movements and that the BCG determined is a measure of the velocity of egg movements. PMID- 9327627 TI - Electromagnetic compatibility aspects of active robotic systems for surgery: the robotic prostatectomy experience. AB - The paper discusses the susceptibility to electromagnetic interference (EMI) of active robots for surgery, which are safety-critical systems. The high EMI environment of an operating room in the presence of an electrosurgical generator is considered. Experience of a surgeon assistant robot for prostatectomies in improving the immunity to EMI is described. It has been found that effective isolation of the robotic system hardware from grounded metal objects provides significant improvements to safety by its immunity to EMI, in minimising the flow of high-frequency current to ground through the system hardware. PMID- 9327628 TI - Precision LED-based stimulator for focal electroretinography. AB - The authors discuss the technical problems commonly encountered in the design of devices used in the functional analysis of the central retina (macula) and its neuronal elements. They present a simple effective solution for introducing some of the most recent and interesting results of neurophysiological and psychophysical research into the eye clinic. PMID- 9327629 TI - "A tissue of the most flagrant anomalies": smallpox vaccination and the centralization of sanitary administration in nineteenth-century London. PMID- 9327630 TI - Vaccination policy of the Faculty of Physicians and Surgeons of Glasgow, 1801 to 1863. PMID- 9327631 TI - Boyle versus the Galenists: a suppressed critique of seventeenth-century medical practice and its significance. PMID- 9327632 TI - The English sweating sickness of 1551: an epidemic anatomized. PMID- 9327633 TI - To treat or not to treat...? PMID- 9327634 TI - Occupational exposure to inorganic mercury vapour and reproductive outcomes. AB - The effect of exposure to inorganic mercury on the pregnant woman and her foetus has received little attention. Transport of elemental inorganic mercury into foetal tissues has been reported, and prior studies indicate a higher incidence of adverse pregnancy outcome. The effects of occupational exposure to inorganic mercury on pregnancy were investigated among 46 exposed women workers: controls were 19 women working in non-production areas of the same factory. There were 104 recorded total pregnancies during the period 1948-77. The study revealed a higher frequency of adverse reproductive outcomes, especially congenital anomalies, among the women exposed to inorganic mercury levels at or substantially lower than 0.6 mg/m3; no significant differences in the stillbirth or miscarriage rates were noted between the two groups of women. The overall foetal death rate in this study was similar to New York state (USA) and national levels for the same period. PMID- 9327635 TI - Occupational risk and toxicology evaluations of industrial water conditioning. AB - This study addresses the chemical and toxicological questions due to the wide use of chemical treatment programmes for industrial cooling water. First, natural problems encountered in cooling tower systems were presented and grouped into three categories: (i) scaling; (ii) corrosion and (iii) biofouling. Chemical solutions adopted in industrial plants were outlined for each one in order to minimize damage and categorized as shut-down, production loss, heat transfer reduction, upsets, etc. Above all, the purpose of the work was to identify the most dangerous chemicals normally used, which means sources of chemical risk for safety workers and their environment; thus, symptoms of exposure, prevention measures and protection tools are also described. PMID- 9327636 TI - Flour protein antigens in occupational flour hypersensitivity. AB - Thirty serum samples from clinical cases of flour hypersensitivity were analyzed for wheat or rye flour protein antibodies. The patients included 20 bakers and 10 others who also had occupational flour exposure. Twenty-three cases had antiflour antibodies which recognized antigens other than control sera in the flour protein patterns. The immunologic response of individual cases seemed very variable in view of the numerous differences between the cases in the antigen-antibody reactions. For the practical purposes, the flour protein antigens were divided in three groups, i.e., those larger than 80 kDa, those between 80 and 50 kDa and those smaller than 50 kDa. Cases with flour-induced dermatitis (n = 8) showed sensitization towards antigens in all size classes while those with rhinitis or asthma showed more antigens with a molecular weight less than 50 kDa. The test offers a possibility to independently verify an exposure to flour while it does not substitute for the conventional immunologic diagnostic tests. PMID- 9327637 TI - Changes in neuropsychological symptoms and moods among tanker drivers exposed to gasoline during a work week. AB - The purpose of this study was to investigate changes in the neuropsychological symptoms and moods among tanker drivers during a work week and the associations of the changes with exposure to gasoline (including 10% methyl-tert-butyl ether). The target group for the study consisted of 101 road tanker drivers from three Finnish oil companies in various parts of Finland. The control group consisted of 100 milk delivery drivers from two milk companies from the same localities. Interviews were conducted before the work week and at the end of the same work week. Standardized symptom questionnaires were used to elicit responses to questions about symptoms and moods. In the questionnaires tanker drivers scored significantly higher in the fatigue scale at the end of the work week than before the work week. Our findings show that tanker drivers with long exposure to gasoline during the work week developed significantly higher changes in fatigue scores than drivers with short exposure. During the exposure situations, 20% of tanker drivers reported acute symptoms (headache, dizziness, nausea, dyspnoea, irritation of saliva excretion) at the end of work week voluntarily. These symptoms have been connected, with MTBE (methyl-tert-butyl ether) exposure, among others. Exposure to MTBE during the work week can be reason for acute symptoms among the tanker drivers in this study. PMID- 9327638 TI - Communication between an occupational physician and other medical practitioners- an audit. AB - Four hundred and seventy-two consecutive referral episodes relating to 386 patients attending the Occupational Health Department of a general teaching hospital were analyzed to evaluate the frequency, content and effect on management of communications between the occupational physician and other doctors. In all, 250 episodes (53%) were associated with such a communication. The likelihood of a communication was strongly influenced by reason for referral, particularly in respect of long or short term sickness absence; univariate odds ratios (OR) = 10.58, 95% CI = 8.13-27.08) and 2.65, 95% CI = 1.55-4.60) respectively; a medical diagnosis of psychiatric illness (OR = 3.17, 95% CI = 1.69-5.97)); and by number of consultations. Communication was also more likely when the occupational outcome was ill health retirement, rehabilitation in work or modified work. Ninety-eight per cent of specific requests for information or an opinion elicited a reply. Information received from other doctors influenced the occupational health physician's management in 52 referral episodes (20%). Specific action by GPs as a result of communication was documented in 54 and by specialists in 37 episodes. The importance of communication between occupational health physician and other doctors in the occupational health process is confirmed. PMID- 9327639 TI - Contamination incidents among doctors and midwives: reasons for non-reporting and knowledge of risks. AB - A 6-month retrospective self-administered questionnaire study of 482 doctors and 380 midwives in two NHS Trusts was undertaken. The response rate was 384 (80%) and 293 (77%) respectively. The study revealed that only nine per cent of doctors and 46% of midwives had reported the contamination incidents they had received. The doctors' main reason for non-reporting was 'too time consuming' and midwives' was 'did not consider anything could be done', although their awareness of the active management of contamination incidents by occupational health departments was good. Seventy-seven per cent of doctors and 69% of midwives underestimated the risk of contracting hepatitis B virus from a needlestick injury, whilst 52% of doctors and 36% of midwives underestimated the risks of acquiring infection with HIV (human immunodeficiency virus) infection following such an injury. Strategies for improving the knowledge of the potential risks of contamination incidents and methods for facilitating ease of reporting are discussed. PMID- 9327641 TI - Genetic screening--uses, potential abuses and ethical issues. AB - Potential uses, abuses and control of genetic screening techniques are discussed and the ethical issues arising from their use are compared with those for existing health screening practices. The main emphasis is upon issues affecting employment but more wide ranging implications are also touched upon. Although the current policy of various bodies is described, the views expressed are personal. PMID- 9327640 TI - Screening questionnaires for bakers' asthma--are they worth the effort? AB - The use of a respiratory screening questionnaire is recommended annually to screen bakery workers in the UK. We compared questionnaire screening with other methods of detecting workers with asthmatic symptoms and then assessed the significance of these symptoms with careful investigation and follow-up. Reasons for questionnaire failures were then explored. A questionnaire was issued to 362 flour-exposed workers in a large bakery. All positive respondents to respiratory symptom questions were interviewed by an occupational nurse. Workers with occupationally related symptoms at this interview were referred to the chest clinic. In addition, workers with negative questionnaires were screened using attendance records, sick notes and direct workplace observations. Workers with frequent absence from work or sick notes with respiratory diagnoses were interviewed in the same manner as those answering the questionnaire positively and then referred to clinic. At clinic, a diagnosis was made for each worker on the basis of clinical assessment, spirometry, serial peak expiratory flow rate (PEFR) analysis and radioallergosorbent testing (RAST) testing for specific IgE. Using the clinic diagnoses, the referral routes were audited to assess the rates of case detection of asthma and occupational asthma. The respiratory screening questionnaire identified 68 workers with respiratory symptoms. Of these, 21 proceeded to full assessment. A diagnosis of asthma was made in five cases, one of which was bakers' asthma. In addition, 11 workers not reporting any symptoms by questionnaire were referred to clinic and five were diagnosed as having asthma. Screening questionnaires may lead to an underestimate of the prevalence of asthmatic symptoms and as such should not be used alone in workplace screening. PMID- 9327642 TI - Ethical issues in occupational medicine practice: knowledge and attitudes of occupational physicians. AB - Views on ethical conduct in occupational medicine practice can vary from country to country and even between occupational health practitioners. However, there are many areas of common agreement, and this is apparent on comparing guidance documents on ethics produced by several different organizations. The usefulness of these documents will depend in part on how aware practitioners are of their existence. A standardized questionnaire administered to 70 occupational physicians in the Netherlands, UK, and Singapore showed that there was a lack of awareness of guidance documents on ethics, even for publications from their own countries. Only five of the 70 respondents consulted an ethics document in the past year. In addition to publications, other avenues were used for advice on ethical issues. There was a difference in opinion between the physicians from Singapore and those from the two European countries on whether specific occupational health activities were ethical. These findings reinforce the need for international guidance on ethics to take into account differences in attitudes and practice between countries. On many issues there was no unanimity of opinion, even between occupational physicians from the same country. This may be an indication of the complexity of ethical matters, and provides a rationale for publishing guidance on ethics in occupational medicine. PMID- 9327644 TI - Data acquisition, people and exposures. PMID- 9327643 TI - SWORD '96: surveillance of work-related and occupational respiratory disease in the UK. AB - Chest and occupational physicians who report to the SWORD surveillance scheme are estimated to have seen some 3,300 new cases of work-related respiratory disease in 1996. This total has regained the level recorded prior to a low in 1995, probably because of improved chest physician participation and the introduction of a sampling system for occupational physicians. Trends in disease incidence have remained fairly constant with some changes only in pneumoconiosis and inhalation accidents. It is of concern that there has been no evidence of a decrease in frequency of occupational asthma or in any of the incriminated agents since the scheme began in 1989. Comparison with other sources of data shows that, for asthma, SWORD records a relatively high frequency in women, a substantial proportion of whom do not appear to receive compensation. For mesothelioma, rates based on death certificates continue for understandable reasons to run at about twice the level reported to SWORD or as reflected by successful claims to the DSS for industrial injuries benefit. The SWORD programme is now one of six clinically based reporting schemes which, by the end of 1997, are planned to cover all types of occupational disease in the UK. PMID- 9327645 TI - Comparison of the respiratory and hemodynamic responses of healthy subjects to exercise in three different protocols. PMID- 9327646 TI - An Advisory Committee Statement (ACS). National Advisory Committee on Immunization (NACI). Supplementary statement on hepatitis prevention. PMID- 9327647 TI - Postmortem cesarean section with infant survival: a case report of an HIV infected patient. AB - Postmortem cesarean sections are rare events, but modern technology is forcing society to explore the definition of viability and the legal rights of both mother and fetus that ultimately will affect the frequency and use of this rare operation. Since the Human Immunodeficiency Virus (HIV) prevalence rate and the number of critically ill obstetrical patients with Acquired Immunodeficiency Syndrome (AIDS) continues to rise, it is reasonable to assume more patients and physicians may be confronted by issues concerning fetal and maternal rights and other considerations in perimortem delivery. A 33-year-old woman, at 27 weeks' gestation, with fulminant AIDS was admitted to the intensive care unit (ICU) in respiratory distress. As her condition deteriorated the complex problem of caring for both mother and fetus emerged. A patient advisory panel explored the issues with the patient, her family, and her health care team. Eventually a peri/postmortem cesarean section was performed on the mother when she suffered an acute fatal cardiorespiratory arrest. With the prevalence of AIDS increasing and with most women not being tested prior to pregnancy, many obstetricians will be confronted with new medical and legal challenges. Establishing comprehensive medical management for the critically ill obstetrical patient and understanding the legal rights of both patients (mother and fetus) will help avoid conflicts and potentially improve survival. PMID- 9327648 TI - Multimodality treatment in the management of esophageal cancer: neoadjuvant chemoradiotherapy followed by transhiatal esophagectomy. AB - Esophageal cancer, although not one of the more common malignancies in the United States, remains a significant problem. Nearly as many patients as are diagnosed die in the same year, regardless of the treatment employed. Surgery is considered the mainstay of therapy. Esophagectomy with the use of the stomach as a substitute is preferred. Radical procedures have not proven more effective in extending survival. Because of the poor five-year survival rate, multimodality therapy with preoperative chemoradiotherapy (neoadjuvant therapy) followed by esophagectomy has shown encouraging results. Two illustrative cases are presented, one with adenocarcinoma and one with a squamous cell carcinoma, that were treated in this manner. PMID- 9327649 TI - Diet and cancer. AB - Unhealthy diets are a major cause of cancer. Extensive scientific research implicates three components of diet as being particularly important in the etiology of various cancers: dietary fat, fruits and vegetables, and fiber. The evidence linking these components to the incidence of numerous cancers warrants physicians making strong dietary recommendations to reduce their patients' risk of developing cancer. PMID- 9327650 TI - Psychotropic medication use in people with developmental disabilities. AB - Psychotropic medications are frequently used to treat undesirable behaviors in persons with developmental disabilities. Successful use of these drugs is dependent on accurate assessment of the psychiatric disorder or behavioral problem. Treatment of aggression and self-injurious behavior and the use of antipsychotics, antidepressants, mood stabilizers, anxiolytics, beta-blocking agents, and naltrexone will be discussed. PMID- 9327652 TI - Broadening our approach among diverse populations. PMID- 9327651 TI - The Muse family of Maryland: to them much was given. PMID- 9327654 TI - The cardiovascular benefits of regular participation in physical activity. PMID- 9327653 TI - Levels of physical activity among Rhode Island adults. PMID- 9327655 TI - Diabetes and exercise. PMID- 9327656 TI - The benefits of physical activity in regard to cancer. PMID- 9327657 TI - Exercise and the skeleton. PMID- 9327658 TI - Physical activity affects weight loss and weight management. PMID- 9327659 TI - The psychological benefits of exercise. PMID- 9327660 TI - Potential adverse effects. PMID- 9327661 TI - Physical limitations to exercise: what they are; how to overcome them. PMID- 9327662 TI - The challenge of behavior change. AB - In a culture that is becoming increasingly automated, motivating individuals to choose to be active remains a great challenge. Recognizing that individuals differ in their motivation to become active and then tailoring the counseling message according to the individual's readiness for change have been shown to help spur behavior change. Developing means for designing interventions for moderate activity and additional recommendations for patient-treatment matching will help to promote physical activity. PMID- 9327663 TI - Physician-delivered physical activity counseling. PMID- 9327664 TI - An ominously low potassium level. PMID- 9327665 TI - Preferred types of physical activity among Rhode Island adults. PMID- 9327666 TI - Femoral pseudoaneurysms in drug addicts. AB - Femoral pseudoaneurysm is a serious complication in drug addicts who habitually inject via the groin. A total of 33 drug addicts presenting with 34 infected femoral pseudoaneurysms were treated in the Department of Surgery, the University of Hong Kong, Queen Mary Hospital from July 1993 to June 1996. There were 27 men and 6 women, with ages ranging from 23 to 76 years (mean 39.6 years). Positive intraoperative tissue cultures were seen in 29 (85%), with 17 being pure growth of methicillin-sensitive Staphylococcus aureus (MSSA). Twenty-four pseudoaneurysms involved the femoral bifurcation and were treated by triple ligation of the common femoral, superficial femoral, and profunda femoris arteries. Seven other limbs underwent ligation of the common femoral artery alone, and three had superficial femoral artery ligation. Nineteen limbs had the external iliac artery ligated in addition to the femoral ligation for better proximal control. The mean postoperative ankle-brachial index (ABI) was 0.43 and 0.52 in those with triple ligation and those with single-vessel ligation, respectively. There was no hospital mortality, and all patients were discharged with a viable limb. The duration of follow-up ranged from 2 to 36 months (mean 15.5 months). Four patients were asymptomatic, but the rest suffered some degree of intermittent claudication. No delayed limb loss was identified. We conclude that systemic antibiotics active against MSSA are the antibiotics of choice in drug addicts with infected femoral pseudoaneurysms. Ligation and excision of the pseudoaneurysm without revascularization is safe, with acceptable morbidity and a low limb loss rate. PMID- 9327667 TI - Sentinel node biopsy in melanoma patients: dynamic lymphoscintigraphy followed by intraoperative gamma probe and vital dye guidance. AB - Biopsy of the first tumor-draining lymph node (sentinel node, SN) is bound to become the procedure of choice in regional staging of melanoma patients. A tumor negative SN virtually excludes lymphatic metastases and obviates the need for lymph node dissection. The aim of this study was to combine the advantages of three known techniques to improve the yield of successful SN biopsies. A total of 150 drainage areas in 135 patients was evaluated. First, preoperative dynamic and static lymphoscintigraphy was performed after injection of technetium 99m colloidal albumin. In all patients one to three focal accumulations, concordant with SNs, were seen in the lymphatic drainage areas, in 97% within 20 minutes from injection of the tracer. Peroperative identification of the SN, 2 to 24 hours after injection of the tracer, was done with a handheld gamma probe to estimate the optimal site for the small incision and to guide preparation. Vital dye was injected just preoperatively and served to facilitate the final identification and biopsy of the SN. A total of 216 SNs were biopsied. Micrometastases were found in 39 SNs in 30 drainage areas, and in 22 of the 30 of the SN was the only node harboring tumor. In 5 of 30 drainage areas, the SN did not contain blue dye and would not have been found without the gamma probe. Up to now (follow-up 233-691 days) no recurrence has developed in the lymphatic drainage areas where the SN was tumor-free. It was concluded that by combining these three techniques the SN could be detected and excised in all patients. The procedure combines a steep learning curve with high sensitivity. PMID- 9327668 TI - Stereotactic breast biopsy as an alternative to excisional biopsy. AB - The current widespread use of modern mammography has increased the detection of suspicious mammographic lesions, which has led to a greater number of diagnostic surgical biopsies. Most of these lesions referred for surgical biopsy have been benign. The reduction in the number of benign surgical biopsies can reduce medical costs and unnecessary invasive breast surgery. Stereotactic large-core needle biopsies in patients with suspicious mammographic lesions can preselect the breast lesions that need further evaluation with surgical excisional biopsy. Our results with stereotactic large-core needle biopsies support this alternative approach to the workup of suspicious mammographic lesions. PMID- 9327669 TI - Long-term prognosis of patients with surgical wound infections. AB - This study examined if surgical wound infections (SWI) result in a severe prognosis regarding general health and increase the consumption of social resources. A group of 1301 patients were interviewed by self-administered questionnaires during 1993-1994, while operated during hospitalization in seven Danish hospitals. These patients were followed up at least once by similar questionnaires at a median time of 5.5 and 10.0 months postoperatively. The consequences of surgically diagnosed SWI were analyzed in a hospital cohort of 58 infected patients and 648 controls. Postdischarge infections were analyzed in a patient cohort of 263 cases and 767 controls. Changes in health was measured by the General Health Questionnaire, Activities of Daily Living index, and self assessed health. Consumption of resources were estimated by reliance on assistance from family and friends, use of home services, and contacts to doctors. It was found that the long-term prognosis of general health was unaffected by SWIs. However significantly increased social dependence was found for patients with SWIs compared to uninfected patients. Almost one-fourth of the operations were complicated by an SWI. Most of the infections were recognized only after discharge and were thought to be of minor importance, which may explain why no chronic impairment of the health was found for patients with an SWI. A bias toward no-effect may have been introduced if patients with severe SWIs abstained from participation. The societal cost of care for patients with minor infections seems to be large. The causal relation between outcome and SWI needs to be further investigated. PMID- 9327670 TI - Patterns of intestinal motility recovery during the early stage following abdominal surgery: clinical and manometric study. AB - The purpose of this study was to evaluate the pattern of recovery of intestinal motility using manometric and clinical assessment in the postoperative ileus following abdominal surgery. We reviewed the charts of 18 patients who underwent partial colectomy for colon cancer (group A, without vagotomy) and compared them to those of 15 patients who underwent gastrectomy with truncal vagotomy and reconstruction by the Billroth I technique (group B, with vagotomy). A three lumen catheter was inserted via the nose into the proximal jejunum for drainage of intestinal fluids and to record intestinal motility at laparotomy. The first appearance of a burst activity following a silent period and a migrating motor complex (MMC) was at 3.8 hours (mean) and 16.4 hours, respectively, in group A, and at 2.5 and 14.3 hours, respectively, in group B. The first audible bowel sounds and the time of feeling abdominal fullness and passage of flatus was at 34.8, 47.5, and 62.9 hours (group A), and at 43.0, 53.8, and 77.9 hours (group B), respectively. The peak volume of nasointestinal drainage was recorded at 16 to 24 hours in group A and 40 to 48 hours in group B (p < 0.05). As measured by manometric examination, an MMC reappeared within a few hours after surgery. Physiologic postoperative ileus was ended when MMCs extended throughout the gastrointestinal (GI) tract with forwarding GI fluids. PMID- 9327671 TI - Antibiotic prophylaxis and open groin hernia repair. AB - Antibiotic prophylaxis is not routinely given for nonimplant, clean operations, although this view has recently been challenged. We have conducted a randomized multicenter, double-blind prospective trial to compare co-amoxiclav with placebo in 619 patients undergoing open groin hernia repair. Altogether 563 (91%) patients fulfilled the protocol; 283 received co-amoxiclav and 280 placebo. There was no difference between the groups in the number of patients receiving local or general anesthetic, the type of repair performed, the use of a subcutaneous fat suture, the type of skin closure used, the use of wound analgesia, or the use of a wound drain. Patients were given a card to return to the hospital in the event of their wound discharging or their needing to see their general practitioner. All patients were reviewed at approximately 6 weeks after operation. Fifty (8.9%) patients sustained a wound infection, 25 in the co-amoxiclav group and 25 in the placebo group. We conclude that antibiotic prophylaxis is of no benefit to patients undergoing open groin hernia repair. PMID- 9327672 TI - Patterns of injury in victims of urban free-falls. AB - The objective of this study was to identify the patterns of injury in urban free fall victims so as to establish guidelines of management. This prospective study at an academic level I trauma center included 187 consecutive patients who presented to our trauma center during a 9-month period (September 1994 to June 1995) after a fall from a height of 5 to 70 feet. Only three falls were from heights of more than 40 feet. Of these patients, 116 (65.1%) suffered significant trauma. Fractures were the most common injuries, accounting for 76.2% of all injuries. Spinal fractures were detected in 37 patients and were associated with neurologic deficits in 7. Intraabdominal injuries occurred in 11 patients, requiring operative intervention in 9 of them. Solid organ lacerations prevailed, but small bowel perforation and bladder rupture were present in one case each. A significant retroperitoneal hematoma was detected in only one case and a thoracic aortic rupture in one more. The height of the fall correlated highly with the incidence of intoxication and severity of injury, the need for operation, the length of hospitalization, and mortality. Most urban free-falls occur from moderate heights. The spinal column is frequently injured and therefore should be thoroughly assessed clinically and radiographically in all fall victims. Intraabdominal organ injuries are much more common than retroperitoneal ones. Thus the abdominal cavity should be the primary target of aggressive workup in hemodynamically unstable patients. The height of the fall is a good predictor of injury severity and outcome prognosis. PMID- 9327673 TI - Radical transhiatal esophagectomy with two-field lymphadenectomy and endodissection for distal esophageal adenocarcinoma. AB - Distal adenocarcinoma of the esophagus is defined as a tumor originating from an endobrachyesophagus or a tumor with its main tumor mass (more than two-thirds) located in the distal tubular esophagus. Controversy exists about the optimal mode of surgical resection. Some favor transthoracic esophagectomy, whereas others prefer transhiatal (blunt) esophagectomy. A radical transhiatal esophagectomy (RTE) combined with two-field lymphadenectomy and mediastinoscopic dissection of the upper thoracic esophagus (endodissection) is described herein. We assessed the short- and long-term results of this technique and compared them to a historical group of patients undergoing conventional transhiatal esophagectomy (THE) for adenocarcinoma of the distal esophagus. Altogether 124 patients underwent transmediastinal esophagectomy because of adenocarcinoma of the distal esophagus in our department between January 1986 and May 1995. Thirteen of these patients were excluded from this analysis because of preoperative chemotherapy. The remaining 109 patients were divided into two groups: 62 patients who underwent THE between January 1986 and March 1991 (51 men, 11 women; mean age 65.3 years, range 31-83 years) and 47 patients who had RTE between April 1991 and May 1995 (44 men, 3 women; mean age 63.4 years, range 41-84 years). To compare the long-term results of RTE and THE, we used a matched pairs analysis considering tumor stage and age. The hospital (30-day) mortality was marginally lower in the RTE group (4.3% versus 6.4%), resulting in an overall mortality of 5.5%. The rate of pulmonary complications was insignificantly lower in the RTE group [19.1% RTE versus 25.8% THE; not significant (NS), and the rate of postoperative cardiac abnormalities significantly decreased after RTE (2.6% RTE versus 19.3% THE; p < 0.05). The overall rate of R0 resections was 87.2% (82.2% RTE, 87.1% THE). Overall survival was similar within the two study groups. Complete tumor removal, T and N stages, and the lymph node ratio were identified as prognostic factors for long-term survival. Overall survival was better after RTE than after conventional THE in patients with involved lymph nodes. The mean number of resected lymph nodes per patient in the RTE group was 26.7. Positive lymph nodes were most common in the paracardial region and at the lesser curvature (72%/10.8% of all invaded abdominal nodes). In the mediastinum positive nodes were most common in the paraesophageal and paraaortal region (48%/27% of all mediastinal nodes). Patients with positive abdominal and mediastinal lymph nodes had a poor long-term prognosis. Distal adenocarcinoma of the esophagus can be safely resected by RTE with two-field lymphadenectomy and endodissection. This technique allows radical "enbloc" resection of the tumor-bearing distal third of the esophagus, which includes the primary area of lymph node metastasis of adenocarcinoma of the distal esophagus: the lower mediastinum and paracardial region. The analysis showed that RTE incurred fewer cardiac complications and a better overall survival in N1-positive patients when compared retrospectively to THE. Intraoperative mediastinoscopy allows controlled dissection of the upper mediastinum and biopsy of several mediastinal lymph nodes, with the advantage of providing additional staging information. PMID- 9327674 TI - Treatment of primary multiple early gastric cancer: from the viewpoint of clinicopathologic features. AB - The treatment of multiple early gastric cancer was investigated through the clinicopathologic assessment of 61 cases of primary multiple early gastric cancer (82 accessory lesions) treated by surgical resection over a 15-year period. These cases accounted for 11.7% of all cases of early gastric cancer resected during the same period. The 61 patients included 48 men (mean age 64 years) and 13 women (61 years). Of the 82 accessory lesions, 41 (50%) were located in the same region as the main lesion. The most frequent combination of macroscopic types of the main lesion and the accessory lesion was depressed type/depressed type (28 cases). The main lesion was of the well differentiated type in 39 (64%) of the 61 cases; the accessory lesion was also well differentiated in 37 of the 39 cases. Of the 82 accessory lesions, 29 (35%) had been overlooked preoperatively; most of them were located in the middle third of the stomach and included 17 depressed and 10 flat lesions, most of which measured no more than 1 cm. Cases of multiple early gastric cancer are characterized by the predominance of male patients of advanced age (> 60 years), a combination of the same macroscopic type of the main and accessory lesions, and well differentiated carcinoma. Lymph node metastasis and vascular invasion are equally or less frequent than in cases of solitary early gastric cancer. The postoperative crude survival rate in patients with multiple gastric cancer is similar to that in those with solitary gastric cancer. Therefore we believe that multiple early gastric cancer does not require extended operative procedures. Endoscopic treatment may be indicated if each lesion fits the criteria for treatment and careful follow-up is ensured. PMID- 9327675 TI - Prognostic impact of splenectomy on gastric cancer: results of the Korean Gastric Cancer Study Group. AB - The relation between splenectomy and survival time after curative total gastrectomy for gastric cancer was reviewed retrospectively on 492 patients treated at nine hospitals between 1989 and 1993. Altogether 260 patients underwent splenectomy, and 232 patients did not. A univariate analysis revealed that the survival time of patients with splenectomy was significantly less than those without splenectomy (p = 0.0265). In a subgroup of our patients stratified to adjust for the stage of disease, there was no significant difference between the survival rates. Splenectomy remained insignificant according to the multivariate analysis using Cox's proportional-hazard regression. The splenectomy group was associated with more risk factors (e.g., T3/T4 tumors, positive nodes, stage greater than III, large tumor size) that are powerful predictors of death due to gastric cancer. In a separate multivariate analysis after eliminating those who had a T4 tumor invasion or a N2 nodal positivity from the analysis (or both), splenectomy again remained insignificant. In conclusion, we could not find any beneficial effect of splenectomy in gastric cancer patients in this retrospective multivariate analysis. We can presume that splenectomy cannot increase the survival rate so long as the splenectomy group has more risk factors than the nonsplenectomy group. Therefore randomized prospective clinical trials using more precise criteria to indicate the need for splenectomy are needed to assess whether splenectomy is beneficial. PMID- 9327676 TI - Surgery for early gastric cancer: a European one-center experience. AB - Between 1972 and 1995 a total of 251 patients with early gastric cancer underwent resection in our department of surgery. At the time of the operation 10.8% of the patients were proved to have lymph node involvement, and two already had distant metastases. A subtotal gastric resection was performed in 59.8% of cases (n = 150), a total gastrectomy in 33.8% (n = 85), and either a proximal or an atypical resection in 6.4% (n = 16). Since 1985 subtotal distal resection and total gastrectomy were accompanied by a systematic lymphadenectomy of compartments I and II. The overall postoperative morbidity was 18.3%, and the hospital mortality, 4.9%; it was only 1.6% within the last decade. Concerning these short term results there were no statistically significant differences between the different surgical procedures. The cumulative overall 5-year-survival rate was 82.6%. There was no statistically significant influence of either the different surgical procedures or the histologic types according to the Japanese classification of early gastric cancer. PMID- 9327677 TI - Nonparasitic cysts of the liver: results and options of surgical treatment. AB - Nonparasitic cysts of the liver (NPHC) are highly variable in respect to appearance and therapeutic approach. The treatment of these cysts varies according to the nature and appearance of the disease. Based on the variable nature of disease and the various therapeutic options, all of which were attempted in our patients, the most suitable mode of treatment for different forms of NPHC are discussed. Ninety-one patients with NPHC who had been treated surgically from 1977 through 1995 were examined retrospectively. Asymptomatic peripheral cysts measuring up to 10 cm do not require further treatment. Computed tomography (CT)-guided aspiration (n = 9) should be regarded as a palliative measure. Within a short period, CT-guided aspiration led to recurrence of symptoms in seven of our patients. Standard treatment of NPHC is fenestration with widest possible excision of the cystic wall, which can be performed laparoscopically (n = 10) or by the conventional surgical mode (n = 54). One patient was initially operated by the laparoscopic technique but developed bleeding, which necessitated conversion to the open mode. Three patients underwent synchronous laparoscopic cholecystectomy. Recurrence rates were similar: 11% in the laparoscopically treated group and 13% in the group that underwent conventional open surgery. Conventional surgical treatment was always successful in cases of solitary cysts. However, in cases of multiple cysts measuring more than 5 cm, conventional surgery was followed by recurrence of symptoms in 26% of patients (7/27), who then had to undergo a second operation. Partial resection of the liver (n = 9) was successfully performed in cases of polycystic disease (n = 5) with concomitant enlargement of the organ as well as in cases of large solitary cysts of the left lobe of the liver (n = 4). In patients in whom we found that the cysts communicated with the ductal system (n = 3), we performed a cystojejunostomy to drain the bile. The complication rate was low. In addition to frequent postoperative ascites, which necessitated no further intervention, we observed infectious complications in four patients. Twenty patients (22%) expired during a mean follow-up period of 6.2 years. Interestingly, deaths were frequently associated with malignancy (11/20). After fenestration of multiple cysts measuring > 5 cm, the patients are at high risk for recurrence. Hence partial resection of the liver is an excellent therapeutic alternative in selected patients with polycystic disease and massive enlargement of the organ in whom the disease could not be controlled by simple fenestration. The results of this study show that laparoscopic fenestration should replace the conventional surgical technique as the gold standard in cases of NPHC because the laparoscopic technique is less stressful for the patient and is associated with a rate of success similar to that of the conventional technique. PMID- 9327678 TI - Intraperitoneal dissemination probably caused by needle biopsy of alveolar echinococcosis of the liver: experimental study. AB - Alveolar echinococcosis of the liver (AEL) is a parasitosis with a potential for malignant tumor-like behavior. The disease is diagnosed by a combination of serologic tests, diagnostic images, and the histology of needle biopsy specimens. It remains unresolved whether the biopsy induces subsequent troubles. We designed this study to investigate critical problems after needle biopsy of AEL lesions using an experimental model. Five samples were prepared from the resected lesions of AEL patients: (A) 10% suspension of trypsin digests of the minced lesion; (B) 10% suspension of mesh-filtered sediment of the minced lesion; (C) 10% sediment suspension after washing the nonminced lesion; (D) supernatant after centrifuging intracystic fluid; (E) 10% sediment suspension after centrifuging intracystic fluid. A 1-ml aliquot of each sample was injected intraperitoneally into jirds (gerbils) or cotton rats, respectively. The animals were sacrificed 12 weeks later, and intraperitoneal metacestodes were observed. All samples except D developed metacestodes, and their histologies were all lesions of typical alveolar echinococcosis. These results suggest that a needle biopsy may cause intraperitoneal dissemination or tracial implantation of the parasites along the track of the needle. PMID- 9327679 TI - Potentially multicentric hepatocellular carcinoma: clinicopathologic characteristics and postoperative prognosis. AB - When multiple hepatic tumors are present, it is sometimes difficult to distinguish between metastatic and multicentric hepatocellular carcinoma (HCC). To identify the important clinicopathologic features of multicentric HCC, we evaluated the clinical characteristics of patients with multicentric HCC and examined the usefulness of surgical treatment in those patients. A total of 99 patients with multiple HCCs were classified into one of the following two groups according to whether their tumors were multicentric or metastatic: Group MO consisted of 18 patients with tumors thought to have developed synchronously from multicentric origins. Group IM consisted of 64 patients with intrahepatic metastases. In this study 18% of the patients with multiple HCCs were thought to have presented with multicentric tumors. This study revealed that synchronous multicentric HCCs often affected multiple segments of the liver and responded relatively well to partial hepatectomy of individual tumor-affected areas. To appropriately treat potentially multicentric HCC, it is important to understand the histopathologic characteristics of multicentric HCC and diagnose during preoperative and intraoperative ultra-sonography, so surgical treatment may be useful. PMID- 9327680 TI - Segment III cholangiojejunostomy for carcinoma of the gallbladder. AB - Jaundice in patients of advanced carcinoma of the gallbladder requires palliation for the distressing symptoms of pruritus and cholangitis. Intrahepatic segment III duct cholangiojejunostomy is a means for alleviating the obstruction with malignant porta block. The authors reviewed their experience with this procedure in 48 patients of carcinoma of the gallbladder. All patients had jaundice; pruritus was present in 44 (92%) and cholangitis in 14 (29%). The level of obstruction was determined preoperatively by percutaneous transhepatic cholangiography. In 32 patients the block was below the level of the bifurcation of the right and left ducts, and 16 patients had a block involving the confluence, isolating the two lobes of the liver. Following segment III cholangiojejunostomy, pruritus was relieved in all and cholangitis in 86% of patients. At the end of 6 weeks a significant fall in serum bilirubin and alkaline phosphatase levels was seen with both types of hilar obstruction. Varying degrees of pain relief was also noted in 75% of patients. Segment III biliary bypass is an effective, one-time, reliable means of palliation for carcinoma of the gallbladder with hilar obstruction. Its efficacy appears to depend on the duration and depth of the jaundice and on the anatomy of the biliary ductal system in the left hemiliver rather than on the type of hilar obstruction. PMID- 9327681 TI - Reduced postoperative morbidity after elective laparoscopic cholecystectomy: stratified matched case-control study. AB - To answer the question whether laparoscopic cholecystectomy (LC) or open cholecystectomy (OC) is safer in terms of complications and to what extent the "learning curve" influences the frequency of complications after LC, we conducted a matched case-control study. First, 200 patients undergoing LC (LC group A), and two groups of 200 patients undergoing LC at two different periods of the learning curve (LC groups B and C) were matched, taking into account sex, age, anesthesiologic risk, and surgical difficulties. We evaluated the frequency and grade of postoperative complications of these patients and of the last 200 patients undergoing OC before the introduction of LC, retrospectively matched with the LC groups. The total rate of complications in the OC group was 16.0% compared with 5.5% in the LC groups (p < 0.003); the difference was particularly significant for complications classified as grade I, in female patients, those younger than 70, those with low anesthesiologic risk (ASA), and those after cholecystectomy without surgical difficulties. Matched case-control analysis revealed that the complication rate in the LC group significantly decreases with experience (P < 0.01). We conclude that LC is today the treatment of choice for symptomatic cholelithiasis and is replacing OC as the gold standard against which new therapies should be compared. PMID- 9327682 TI - Peripancreatic fluid cytology: detection of early rejection versus graft pancreatitis after canine pancreatic transplantation. AB - It is particularly difficult to distinguish between early rejection and graft pancreatitis when early rejection produces an elevated serum amylase level. In this study we determined whether peripancreatic fluid cytology (PFC) can differentiate early acute rejection and graft pancreatitis as an alternative diagnostic tool to graft biopsy that has the potential of pancreatic fistula and hemorrhage. Sixty-two dogs received either a segmental pancreas allograft (n = 25) or autograft (n = 37) heterotopically in the neck. This study included five groups: allografts without immunosuppression (group A, n = 12), allografts with immunosuppression (group B, n = 13), autografts without immunosuppression (group C, n = 11), autografts with immunosuppression (group D, n = 12), and autografts treated by 45 minutes of pretransplant warm ischemia to induce acute graft pancreatitis (group E, n = 14). A closed suction drainage catheter was placed next to the graft to collect peripancreatic fluid daily after the transplant. PFC was performed using May-Gruenwald-Giemsa double-staining technique and compared to the corresponding histology through the observation period. In analyses of 50 functioning grafts, PFC performed on day 1 showed similar neutrophil accumulations in all groups. In sharp contrast, on days 3 and 6, group A had dramatically increased mononuclear cell concentrations in PFC, whereas groups B, C, and D showed significantly lower concentrations, the percent of mononuclear cells among total leukocytes being 47.3 +/- 23.4%, 11.8 +/- 4.9%, 4.3 +/- 1.8%, and 6.4 +/- 2.4% (day 3); and 32.7 +/- 9.8%, 10.5 +/- 4.8%, 7.2 +/- 4.2%, and 8.6 +/- 6.4% (day 6) in groups A, B, C, and D, respectively. On the other hand, in group E numerous degenerating neutrophils with a marked to moderate increase in necrotic tissue fragments were observed by PFC on days 3 and 6. In terms of graft histology on days 3 and 6, group A showed interstitial mononuclear cell infiltration indicating an acute rejection process, whereas groups B, C, and D had minimal inflammatory cell infiltration. In group E graft pancreatitis was histologically confirmed on days 3 and 6. These results suggest that PFC after pancreas transplantation could be a safe, simple, useful diagnostic tool for discriminating early graft rejection from graft pancreatitis. PMID- 9327683 TI - Absorption kinetics of rectal formalin instillation. AB - Formalin instillation has become an accepted treatment of radiation-induced hemorrhagic cystitis and proctitis since the initial report by Brown in 1969 (Med. J. Aust. 1:23, 1969). Although its use is widespread, no studies have been performed to determine the safest, volume or duration of formalin exposure. The purpose of our study was to determine the optimum technique for instillation and the safety margin regarding the maximum time that formalin can be in contact with the rectal mucosa without causing serum toxicity. In a pilot canine study, 4% neutral buffered formalin was instilled into the rectum in 30 ml aliquots for 60 seconds each after which each aliquot was withdrawn; a total volume of 400 ml was used. Our subsequent experiment involved rectal instillation of a single formalin bolus of 100 ml for 1 hour without removal during this time. Formalin metabolites were measured in the blood and urine to assess toxicity. Results indicate that with the latter technique serum formic acid reaches toxic levels within 15 minutes of instillation and may stay elevated for several hours. Metabolites in the urine similarly increase within 15 minutes, lagging only shortly behind the rise in serum levels. Performing formalin instillation in a series of 30 ml aliquots appears to be a safer treatment, as toxic serum levels were not reached and their slight rise above baseline returned to normal within 3 hours. PMID- 9327684 TI - History of perforated duodenal and gastric ulcers. AB - Perforated peptic ulcer as a disease entity has been known since 167 BC. Surgical and nonsurgical treatment strategies for perforated peptic ulcer disease were not developed until the latter half of the nineteenth century. The history of the gradual evolution of the various forms of treatment adopted for the conditions over the last century and a half is described. PMID- 9327685 TI - Report of the study on reducing the occupational risk of infections for the surgeon. PMID- 9327686 TI - Health after coronary stenting or balloon angioplasty: results from the Stent Restenosis Study. AB - This study was designed to compare health-related quality of life (HRQOL) in patients undergoing coronary stenting or balloon angioplasty in the randomized Stent Restenosis Study. The study sample was drawn from patients at nine U.S. clinical sites of the Stent Restenosis Study, a randomized trial comparing Palmaz Schatz coronary stent implantation with conventional balloon angioplasty. One hundred ninety-nine consecutive patients were sent surveys 6 to 18 months after enrollment and 160 (80%) were returned. The survey sent to the patients included the Medical Outcomes Study 36-Item Short-Form Health Survey, the Canadian Cardiovascular Society Classification, and the Duke Activity Status Index. Although patients who underwent stenting had less angiographic restenosis and a tendency for fewer ischemic events, there were few differences in HRQOL after a mean of 456 days after randomization. The group that underwent stenting reported significantly less bodily pain than the group that underwent angioplasty (p = 0.02). Otherwise, there were no significant differences in generic or disease specific measures. In a rating of their overall health, 47% of the group that underwent stenting and 45% of the group that underwent percutaneous transluminal coronary angioplasty reported very good or excellent health. In each group, 60% of the patients reported being symptom free from a cardiovascular perspective. This survey revealed no marked differences in long-term HRQOL between patients who underwent Palmaz-Schatz coronary stenting compared with those who underwent conventional angioplasty. PMID- 9327687 TI - Directional coronary atherectomy for the treatment of acute myocardial infarction. AB - Directional coronary atherectomy (DCA) was performed after intracoronary thrombolysis in 32 patients with a first acute myocardial infarction. DCA was successful in 31 (97%) of 32 patients. Abrupt closure of the treated segment occurred in one patient but was managed successfully by conventional balloon angioplasty. Repeat angiography was performed in 32 patients before discharge (2.7 +/- 0.7 weeks later) and in 29 patients during the follow-up (4.5 +/- 1.5 months later). No restenosis (stenosis > 50%) occurred before discharge; however restenosis occurred in 12 (41%) of 29 patients during follow-up. The restenosis rate in patients with subintimal resection was significantly higher than in those with intimal resection (78% vs 25%, p < 0.01). These data suggest that DCA in patients with acute myocardial infarction is feasible for persistent early patency of the infarct-related coronary artery, but late restenosis continues to limit success and subintimal resection may increase the restenosis rate during the follow-up. PMID- 9327688 TI - Relation between injections before 90-minute angiography and coronary patency: results of the thrombolysis in myocardial infarction 4 trial. AB - The current goal of thrombolytic therapy is to achieve both full (Thrombolysis in Myocardial Infarction [TIMI] grade 3) and early reperfusion. Newer reperfusion strategies may now achieve a high degree of reperfusion even earlier than the traditional 90-minute end point. To determine whether injections before 90 minutes affect this traditional end point, the relation between the number of injections before 90-minute angiography and patency was examined in the TIMI 4 trial. The number of injections before 90-minute angiography was no different between occluded arteries (TIMI grade 0/1 flow) (2.46 +/- 1.78; n = 94) and patent arteries (TIMI grade 2/3 flow) (2.71 +/- 2.42; n = 295) (p = 0.24). The incidence of any injections before 90 minutes was no different in patent versus closed arteries (80.6% [77/98] vs 72.4% [22/304]; p = 0.10). The number of injections before 90 minutes was insignificantly smaller in patients with TIMI grade 3 flow (2.53 +/- 2.53 [n = 184] vs 2.76 +/- 2.03 [n = 204]; p = 0.31), but the incidence of any injections before 90 minutes was significantly smaller in patients with TIMI grade 3 flow (68.8% [132/192] vs 79.5% [167/210]; p = 0.01). No relation was identified between the number of injections before 90-minute angiography and patency at this traditional time point. This observation justifies the judicious use of a limited number of "earlier snapshots" of the infarct-related artery before 90 minutes to ascertain just how rapidly newer thrombolytic regimens achieve patency. Patients with TIMI grade 3 flow had a slightly lower incidence of injections before 90 minutes, perhaps because they did not require as urgent a definition of coronary anatomy. PMID- 9327689 TI - Initial experience with long coronary stents: the changing practice of coronary angioplasty. AB - The initial experience with the use of long coronary stents (> 30 mm in length) was analyzed retrospectively. Sixty-seven stents were deployed in 58 narrowings in 57 patients (34 AVE Microstents, 16 Nir stents, four Gianturco-Roubin II stents, and 13 Wallstents). Stents were implanted in 22 patients with unstable angina, 34 patients with stable angina, and one patient during direct angioplasty for acute myocardial infarction. Eighteen additional short stents were implanted to cover the entire length of the lesions so that an average of one and a half stents were deployed per patient. The length of the narrowings before stenting was 40 +/- 20 mm and the length of the stented segments was 45 +/- 20 mm. Stents were deployed for "bailout" in 23 narrowings, to improve suboptimal results of balloon angioplasty in 18 narrowings, and electively in 17 narrowings. Twenty of the 67 long stents were deployed in saphenous vein grafts. The success rate of stent implantation was 100%. One patient had a rupture of a saphenous vein graft after deployment of two long stents, with tamponade treated by emergency surgery. One patient had chest pain 18 hours after stent deployment; by the time he arrived at the catheterization laboratory the pain had subsided and the angiogram revealed a patent artery with normal flow. There were no other major complications during the hospital course and 1-month follow-up. We conclude that long coronary stents can be deployed successfully in native coronary arteries and vein grafts. They are useful for elective implantation and extremely helpful in bailout situations. The immediate results are excellent, but long-term outcome is awaited. PMID- 9327690 TI - Abrupt vessel closure: changing importance, management, and consequences. AB - Percutaneous coronary interventions have been performed for 20 years. Despite the success and progress of these interventions, abrupt vessel closure has been a dramatic adverse event of coronary interventions. Closure has frequently led to the major complications of death, myocardial infarction, and emergency coronary artery bypass. Because of the fear of this adverse event and its subsequent complications, the applicability of coronary interventions is sometimes limited. The pathologic characteristics of abrupt vessel closure have been recognized as predominantly caused by dissection, with vessel recoil and thrombus formation playing important secondary roles. The recognition of the lesions at risk for abrupt vessel closure has led to a strategy of lesion-specific device therapy to reduce complications. Similarly the role of antiplatelet and antithrombotic therapies have reduced complications. The earliest methods of dealing with abrupt closure was emergency coronary artery bypass surgery with significant rates of morbidity and mortality. With the advent of second-generation devices and techniques, particularly stents, the management of abrupt vessel closure has been simplified and alternatives to emergency coronary bypass are more available. This article will review the history and current status of the prevention and management of abrupt vessel closure and demonstrate that anticipation and management of this complication have been facilitated with reduction of subsequent complications and increased applicability of coronary interventions. PMID- 9327691 TI - Incidence of cardiac sarcoidosis in Japanese patients with high-degree atrioventricular block. AB - In Japan the majority of sarcoidosis-related deaths are due to cardiac sarcoidosis. One of the most common electrocardiographic abnormalities in patients with this disease is atrioventricular block. This study surveyed the incidence of cardiac sarcoidosis in Japanese patients (40 men and 49 women; mean age, 69.1 years) with high-degree atrioventricular block who were admitted to the hospital to receive a permanent pacemaker. We excluded cases in which sarcoidosis had been diagnosed from the involvement of other organs. Patients with the characteristic signs of sarcoidosis underwent echocardiography, radionuclide imaging, and biopsy. Ten cases (11.2%) of cardiac sarcoidosis were diagnosed, most frequently in women aged 40 to 69 years (8 of 25, 32%). Thus the possibility of cardiac sarcoidosis should be carefully considered in middle-aged or elderly Japanese women who show high-degree atrioventricular block. PMID- 9327692 TI - Atrioventricular node reentrant tachycardia in patients with a long fast pathway effective refractory period: clinical features, electrophysiologic characteristics, and results of radiofrequency ablation. AB - Twenty-two patients (group 1) with AV node reentrant tachycardia and a baseline fast pathway effective refractory period (ERP) > or = 500 msec were compared with 30 consecutive patients (group 2) with AV node reentrant tachycardia and a fast pathway ERP < 500 msec. Both groups underwent slow pathway ablation. In the patients with complete elimination of slow pathway, the fast pathway ERP and shortest 1:1 conduction cycle length shortened significantly after ablation but was greater in group 1 (n = 14) than in group 2 (n = 21) (125 +/- 78 msec vs 48 +/- 29 msec, p < 0.001 and 103 +/- 72 msec vs 52 +/- 30 msec, p < 0.001, respectively). In group 1, the shortening of fast pathway ERP was correlated to baseline difference between anterograde fast and anterograde slow ERP (r = 0.806, p < 0.001, slope = 1.08), and the shortening of fast pathway shortest 1:1 conduction cycle length was correlated to baseline difference between anterograde fast and anterograde slow shortest 1:1 conduction cycle length (r = 0.885, p < 0.001, slope = 1.47). During follow-up bradycardia did not develop in any patient and no one required pacing. This shortening of the fast pathway ERP and shortest 1:1 conduction cycle length after complete elimination of slow pathway reduced the concern of subsequent impairment of AV node conduction. PMID- 9327693 TI - Effect of beta-adrenergic stimulation on the QRS duration of the signal-averaged electrocardiogram. PMID- 9327694 TI - Effect of coronary angioplasty on QT dispersion. AB - Increased QT dispersion (QTmax-QTmin [QTd]) reflects inhomogeneous ventricular repolarization that may provide a substrate for serious arrhythmias and is associated with adverse clinical outcomes in patients with heart disease. Effective treatment of acute myocardial infarction or ventricular arrhythmias may reduce QTd, but the effect of coronary revascularization on QTd in patients without these conditions is unknown. In this study, QTd was measured before and 4 and 24 hours after successful angioplasty in 94 patients without ongoing symptomatic myocardial ischemia or malignant arrhythmias. QTd decreased from 434 +/- 17 msec before angioplasty to 354 +/- 15 msec 4 hours (p < 0.05) and 33 +/- 14 msec 24 hours after angioplasty (p < 0.05). QTd was improved in 64% of patients, worse in 28%, and unchanged in 8%. Thus angioplasty significantly improves QTd. This may reflect increased myocardial perfusion and may be inherently beneficial by reducing the propensity for arrhythmias. PMID- 9327695 TI - Effects of beta-adrenergic blockade on dispersion of ventricular repolarization in newborn infants with prolonged QT interval. AB - A prolonged QT interval in the neonatal period and during infancy is associated with higher mortality rates, independently of the appearance of symptoms. This suggests the opportunity of a prophylaxis with beta-blockers in this population. QT interval dispersion is a useful clinical tool to predict the efficacy of antiadrenergic therapy. However, the effects of antiadrenergic interventions on QT interval and QT interval dispersion in newborns are not known. Standard 12 lead electrocardiograms were recorded in 14 newborns with prolonged QT interval, before and after oral administration of 2 mg/kg propranolol. Two electrocardiograms were also recorded in 14 newborns with normal QT intervals, matched for age and sex. In the control group no differences in heart rate, mean QTc, and QTc dispersion between the two recordings were observed. In the newborns with prolonged QT interval, propranolol did not change mean Qtc (from 467 +/- 21 to 451 +/- 26 msec; difference not significant), whereas it decreased spatial QTc dispersion, measured as the difference between the longest and shortest QTc in 12 different leads (from 69 +/- 23 to 42 +/- 13 msec; -39%; p = 0.001). This reduction was explained mainly by a shortening of the maximum QTc (from 504 +/- 25 to 476 +/- 18 msec; p = 0.005), with no change in the minimum QTc, and was not dependent on the slight change in mean QTc, as shown by the decrease in the coefficient of variation of QTc (standard deviation of QTc/mean QTc x 100) from 6.0 +/- 2.2 to 3.3 +/- 0.8 (p = 0.002). Of note, after propranolol, both measures of QTc dispersion reached the same levels observed in the control newborns. During the follow-up (> 2 years for nine of 14 infants), none of the infants had symptoms or arrhythmias. These results suggest that beta-adrenergic blockade with propranolol slightly affects mean QTc, but it significantly decreases the spatial dispersion of ventricular repolarization in newborn infants with a prolonged QT interval. This effect might modify the arrhythmogenic substrate, leading to a reduction of the susceptibility to life-threatening arrhythmias. PMID- 9327696 TI - Stability over time of circadian rhythm of variability of heart rate in patients with stable coronary artery disease. AB - Reproducibility of circadian rhythm of variability in heart rate was studied in 40 patients with stable coronary artery disease who underwent 48-hour ambulatory electrocardiographic recordings at baseline (time 1) and after 4 months (time 2). The standard deviation of the R-R interval and the low-frequency (0.04 to 0.15 Hz) and high-frequency (0.15 to 0.45 Hz) components of variability in heart rate were assessed every 5 minutes. In 35 patients a significant circadian rhythm was observed at both time 1 and time 2 in the standard deviation of the R-R interval, with the acrophase occurring at around 5:00 AM, in the high-frequency amplitude with the acrophase around 3:00 AM, and in the low-frequency/high-frequency ratio with the acrophase around noon. In these patients, parameters of circadian rhythm (mesor, amplitude, and acrophase) showed good within-individual reproducibility with an intraclass correlation coefficient of 0.63 to 0.95 (p < 0.001 for all). In the patients who showed inconsistency about the significance of circadian rhythm between time 1 or time 2, the amplitude of circadian rhythm, even if significant, was found in the lowest five values in the distribution. We conclude that the circadian rhythms of cardiac autonomic activity are stable over time within individual patients with stable coronary artery disease. PMID- 9327697 TI - Patient characteristics and underlying heart disease as predictors of recurrent atrial fibrillation after internal and external cardioversion in patients treated with oral sotalol. AB - The aim of this study was to identify predictors for recurrent atrial fibrillation after internal and external cardioversion in 157 patients. After cardioversion, patients were treated orally with sotalol (174 +/- 54 mg/day). Univariate predictors for recurrence included coronary artery disease (p < 0.05) and advanced age (p < 0.05). Multivariate adjusted risk for relapse increased with the presence of coronary artery disease (odds ratio [OR] 3.4, 95% confidence interval [CI] 1.6 to 8.0), presence of atrial fibrillation > 2 months before cardioversion (OR 2.3; 95% CI 1.4 to 4.5), left atrial diameter > 60 mm (OR 2.1; 95% CI 1.2 to 3.1), and age > 65 years (OR 1.6; 95% CI 1.3 to 3.3). In 26% of patients with lone atrial fibrillation, recurrence was observed compared with 51% of patients with underlying structural heart disease (p < 0.05). The mode of conversion, internal or external, had no impact on the recurrence rate. These findings might be useful for selection of the most appropriate therapy for the individual patient. PMID- 9327698 TI - Right and left ventricular diastolic function in patients with and without heart failure: effect of age, sex, heart rate, and respiration on Doppler-derived measurements. AB - Doppler echocardiography is widely used to assess right and left ventricular diastolic function. Although the factors affecting Doppler-derived measurements have been described in unaffected patients, there is little information available for patients with heart failure. Therefore we performed two-dimensional Doppler echocardiography studies of right and left ventricular function in 140 subjects, 88 with congestive heart failure and 52 age-matched normal subjects. The separate effects of age, sex, heart rate, and respiration were assessed by correlation analysis and multiple linear regression. In normal subjects both left and right ventricular parameters significantly correlated with age and heart rate. No significant effect of respiration was apparent in left ventricular function, but in the right ventricle inspiration caused a significant increase in transtricuspid peak E-wave velocity, E/A ratio, and shortening of the E-wave deceleration time. There was a significant correlation between left and right ventricular diastolic parameters. In heart failure, age correlated weakly with mitral valve peak A wave (r = 0.23, p = 0.03) and with tricuspid valve peak E wave velocity (r = 0.37, p < 0.001), although in those with a restrictive filling pattern age was associated with a significant increased shortening of the mitral E-wave deceleration time (r = -0.8; p = 0.0015). Heart rate and deceleration time of mitral and tricuspid E wave significantly correlated, but heart rate did not correlate with either mitral or tricuspid peak E-wave or A-wave velocities or E/A ratio. In heart failure the effect of respiration was similar to normal subjects. Sex was not associated with Doppler variables in either normals or heart failure subjects. Thus this study demonstrates that age, heart rate, and respiration have important and separate associations with Doppler echo diastolic parameters of the right and left ventricle in normal subjects and similar, although weaker relations in patients with heart failure. PMID- 9327699 TI - Comparison of the degree of hemodynamic tolerance during intravenous infusion of nitroglycerin versus nicorandil in patients with congestive heart failure. AB - BACKGROUND: Continuous exposure to organic nitrates is associated with substantial tachyphylaxis. This study compares the development of tolerance during continuous intravenous treatment with nitroglycerin versus nicorandil over a 24-hour period. METHODS AND RESULTS: Twenty patients with congestive heart failure and pulmonary capillary wedge pressure (PCWP) > or = 18 mm Hg were randomly assigned to nitroglycerin or nicorandil in a double-blind, crossover study. Doses were titrated to obtain a reduction of PCWP of at least 30% and then maintained. The mean pretreatment PCWP for nitroglycerin was 25.4 +/- 6.7 mm Hg, decreasing to 19.0 +/- 6.8 mm Hg at 24 hours. The values for nicorandil were 24.3 +/- 6.3 mm Hg and 15.6 +/- 4.5 mm Hg, respectively. Between-treatment difference was significant (p < 0.01). The difference between the minimal PCWP value and the 24-hour PCWP value for nitroglycerin was 5.1 mm Hg vs 1.4 mm Hg for nicorandil (p < 0.005). The mean systemic vascular resistance was 1418 +/- 355 dynes.sec.cm-5 before nitroglycerin infusion, decreasing to 1312 +/- 353 dynes.sec.cm-5 at 24 hours. Corresponding values for nicorandil were 1420 +/- 366 dynes.sec.cm-5 and 967 +/- 274 dynes.sec.cm-5. Between-treatment difference was significant (p = 0.005). Tachyphylaxis developed in 12 (60%) patients during nitroglycerin infusion versus three patients (15%) during nicorandil infusion. CONCLUSION: This study demonstrates that intravenous nicorandil administration results in significantly less hemodynamic tolerance over a 24-hour period compared with nitroglycerin. This finding may represent a clinical advantage for nicorandil in the short-term treatment of patients with congestive heart failure. PMID- 9327700 TI - Evaluation of plasma natriuretic peptides as markers for left ventricular dysfunction. AB - To test the hypothesis that elevated plasma levels of natriuretic peptides may serve to identify patients with left ventricular (LV) dysfunction, we assessed the predictive diagnostic value of natriuretic peptide levels, in addition to clinical and electro-cardiographic risk factors, as noninvasive indicators of cardiac dysfunction. Plasma levels of atrial natriuretic peptide (cANP) (99-126), N-terminal fragment of proANP (nANP) (26-55), nANP(80-96), brain natriuretic peptide (BNP-32), proBNP(22-46), and C-type natriuretic peptide (CNP-22) were measured in 211 subjects before cardiac catheterization. The strongest correlations with parameters of LV function were found for nANP(80-96) (up to r = -0.55, p < 0.0001), whereas there was no significant correlation with proBNP(22 46) or CNP-22. In patients with LV ejection fractions (LVEF) < or = 45% (n = 38) nANP(26-55), nANP(80-96), cANP(99-126), and BNP-32 were significantly increased (p < 0.001). Partition values for elevated versus normal natriuretic peptide levels were obtained from normal controls and used to separate subjects with and without LV dysfunction. Receiver operating characteristic analysis for LVEF < or = 45% indicated a significantly better diagnostic accuracy for high levels of nANP(80-96), nANP(22-56), cANP(99-126), and BNP-32 than for proBNP and CNP-22. Multivariate analysis by logistic regression identified Q waves and bundle branch block in the electrocardiogram as well as elevated plasma levels of cANP, nANP(80 96), and nANP(26-55) as the strongest independent predictors of low ejection fractions. The relative risk of LV dysfunction was raised up to tenfold in subjects with high natriuretic peptide levels (p < 0.001). The addition of nANP(80-96) and nANP(26-55) to the combination of clinical and electrocardiographic risk factors did not further improve the diagnostic sensitivity for the detection of LVEF < or = 45%, but it markedly increased the overall accuracy (59% to 81%, p < 0.001) and specificity (55% to 81%, p < 0.001). Among natriuretic peptides, elevated nANP(80-96) and nANP(26-55) levels have the strongest impact on the detection of LV dysfunction. They add to the diagnostic information contained in clinical and electrocardiographic factors. Plasma levels alone or in combination with clinical factors seem to be of value for a refined identification of abnormal LV function in the individual patient. PMID- 9327701 TI - Prediction of pulmonary capillary wedge pressure and assessment of stroke volume by noninvasive impedance cardiography. AB - Early recognition of heart failure is important because early treatment reduces mortality and hospitalization rates. In screening for this disease, there is a need for a simple, safe, and cost-effective method to obtain cardiovascular variables. Therefore we developed a noninvasive impedance cardiographic method to predict the pulmonary capillary wedge pressure (PCWP) from the impedance cardiogram. The impedance cardiographic technique, though, was originally designed for stroke volume (SV) determination. The objectives of this study were to validate both variables by comparison with the paired, invasively obtained equivalents. PCWP, measured with a pulmonary artery catheter, was related to the O/C ratio from the impedance cardiogram. The O/C ratio was calculated as the amplitude of the impedance cardiogram during diastole (O) divided by the maximum height during systole (C). Stroke volume was also calculated from the impedance cardiogram according to the equation of Kubicek (SVIC) and compared with thermodilution (SVTD). Data analysis was performed in 24 stable patients who underwent diagnostic heart catheterization. Linear regression analysis showed that the O/C ratio was strongly related to the invasively measured PCWP over a range of 3 to 30 mm Hg (r = 0.92, standard error of the estimate, 3.2 mm Hg). Between SVIC and SVTD a moderate correlation was established (r = 0.69), but after exclusion of the data from patients with an aortic valve disorder (n = 5), the correlation increased considerably (r = 0.87). No significant differences between SVIC and SVTD were found (mean difference +/- 2 SD = 1.8 +/- 28.8 ml). These preliminary observations suggest that impedance cardiography can predict PCWP and measure SV over a wide range of clinically relevant values. The combined measurement of SV and PCWP by impedance cardiography might be a clinical useful tool in screening for heart failure. PMID- 9327702 TI - Comparison between iodine 123 metaiodobenzylguanidine scintigraphy and heart rate variability for the assessment of cardiac sympathetic activity in mild to moderate heart failure. AB - This study demonstrates that in patients with mild to moderate heart failure, cardiac metaiodobenzylguanidine (MIBG) washout positively correlates with normalized low-frequency power in the heart rate variability spectrum. Alterations of the cardiac sympathetic nervous system could be detected with MIBG scintigraphy in patients with normal plasma norepinephrine levels. Therefore cardiac MIBG washout may be a valuable noninvasive technique to assess early alterations in cardiac sympathetic activity that may have potential clinical implications in patients with mild to moderate heart failure. PMID- 9327703 TI - Assessment of autonomic function at rest and during tilt testing in patients with vasovagal syncope. AB - This study evaluated autonomic nervous system function in 30 patients with syncope and a positive tilt test result, 20 with a negative test result, and 20 healthy controls. Indexes of heart rate variability were measured during the intervals immediately before and after tilt, while all subjects were asymptomatic, and over a 24-hour period. There were no significant differences among the groups in any of the indexes of heart rate variability over the 24-hour period. In patients with a positive tilt result, tilting caused a decrease in low frequency (LF) and high-frequency (HF) bands, although the LF/HF ratio did not significantly change. In patients with a negative tilt result there was a decrease in the HF band but no other significant changes. In the controls there was an increase in the LF band and LF/HF ratio and a decrease in the HF band. Our findings showed that patients with vasovagal syncope have no chronic differences from normal subjects in autonomic nervous system activity, but that these patients respond differently to the orthostatic stimulus. PMID- 9327704 TI - Effects of magnetic resonance imaging on cardiac pacemakers and electrodes. AB - In phantom studies we investigated the effects of magnetic resonance imaging (MRI) on pacemakers and electrodes. Twenty-five electrodes were exposed to MRI in a 1.5T scanner with continuous registration of the temperature at the electrode tip. Eleven pacemakers (five single chamber and six dual chamber) were exposed to MRI. Pacemaker output was monitored to detect malfunction in VOO/DOO and VVI/DDD modes. A temperature increase at the electrode tip of up to 63.1 degrees C was observed during 90 seconds of scanning. In seven electrodes the temperature increase exceeded 15 degrees C. Although no pacemaker malfunctions were observed in asynchronous pacing mode (VOO/DOO), inhibition and rapid pacing were observed during spin-echo imaging if the pacemakers were set to VVI or DDD mode. Pacemaker function was not impaired during scanning with gradient-echo sequences. Next to pacemaker dysfunction, electrode heating has to be considered a possible adverse effect when exposing patients with pacemakers to MRI. PMID- 9327705 TI - Frequency, duration, magnitude, and consequences of myocardial ischemia during intracoronary ultrasonography. AB - To determine the frequency, duration, magnitude, and possible adverse effects of ischemia during intracoronary ultrasonography, real-time standard 12-lead electrocardiograms were recorded before, during, and after ultrasonography. Ischemia was defined as new-onset ST segment deviation of > or = 1 mm in one or more leads, measured at J + 80 msec. The magnitude of ischemia was expressed as the sum of absolute ST segment deviations across 12 leads. Eighteen (67%) of 27 patients had ischemia during intracoronary ultrasonography. The electrocardiogram resembled the characteristic pattern observed with occlusion of the vessel under study, involving ST segment elevation in contiguous leads in 89% of patients. A higher proportion of women (88%) had ischemia than men (58%), and women had smaller arterial lumenal areas compared with men (6.3 vs 9.1 mm2; p < 0.05). Individuals with ischemia were smaller than those without ischemia (body surface area = 1.99 vs 1.79 m2; p = 0.01). The mean duration of ischemia was 4 minutes and the mean 12-lead ST segment deviation score was 8.5 mm (maximum 20.5 mm). No patient with ischemia during ultrasonography had complications. Ischemia is common during intracoronary ultrasonography, particularly in women and individuals with smaller vessels; however, no adverse outcomes occur as a result. PMID- 9327706 TI - Echocardiography Doppler in pulmonary embolism: right ventricular dysfunction as a predictor of mortality rate. AB - To test the hypothesis that right ventricular (RV) systolic dysfunction at the time of diagnosis of pulmonary embolism (PE) is a predictor of mortality rate, 126 consecutive patients with PE were examined with echocardiography Doppler (ED) on the day of diagnosis. RV function was assessed by evaluation of wall motion on a four-point scale. The material was divided into two groups: group A (n = 56) with normal or slightly reduced RV function and group B (n = 70) with moderately or severely reduced RV function. The overall mortality rate was 7.9% in the hospital and 15.1% within 1 year. Four deaths occurred in group A and 15 in group B (p = 0.04). All in-hospital deaths (n = 10) occurred in group B (p = 0.002). The variables associated with mortality rate were RV dysfunction and cancer (in hospital, p = 0.002 and 0.004; 1 year, p = 0.04 and < 0.001, respectively). Nine (7.1%) deaths (all in-hospital) were caused by PE. Five of these patients had advanced-stage cancer. The in-hospital mortality rate in patients without cancer was 4%, all from PE and all in group B. In conclusion, RV dysfunction when diagnosis of PE is established is associated with mortality rate. A strategy for risk stratification of patients with PE with ED may be of clinical usefulness. PMID- 9327707 TI - The effect of lead selection on traditional and heart rate-adjusted ST segment analysis in the detection of coronary artery disease during exercise testing. AB - Several methods of heart rate-adjusted ST segment (ST/HR) analysis have been suggested to improve the diagnostic accuracy of exercise electrocardiography in the identification of coronary artery disease compared with traditional ST segment analysis. However, no comprehensive comparison of these methods on a lead by-lead basis in all 12 electrocardiographic leads has been reported. This article compares the diagnostic performances of ST/HR hysteresis, ST/HR index, ST segment depression 3 minutes after recovery from exercise, and ST segment depression at peak exercise in a study population of 128 patients with angiographically proved coronary artery disease and 189 patients with a low likelihood of the disease. The methods were determined in each lead of the Mason Likar modification of the standard 12-lead exercise electrocardiogram for each patient. The ST/HR hysteresis, ST/HR index, ST segment depression 3 minutes after recovery from exercise, and ST segment depression at peak exercise achieved more than 85% area under the receiver-operating characteristic curve in nine, none, three, and one of the 12 standard leads, respectively. The diagnostic performance of ST/HR hysteresis was significantly superior in each lead, with the exception of leads a VL and V1. Examination of individual leads in each study method revealed the high diagnostic performance of leads I and -aVR, indicating that the importance of these leads has been undervalued. In conclusion, the results indicate that when traditional ST segment analysis is used for the detection of coronary artery disease, more attention should be paid to the leads chosen for analysis, and lead-specific cut points should be applied. On the other hand, ST/HR hysteresis, which integrates the ST/HR depression of the exercise and recovery phases, seems to be relatively insensitive to the lead selection and significantly increases the diagnostic performance of exercise electrocardiography in the detection of coronary artery disease. PMID- 9327708 TI - Natural history and serial morphology of aortic intramural hematoma: a novel variant of aortic dissection. AB - BACKGROUND: Acute aortic dissection is a cardiovascular emergency that requires prompt diagnosis and treatment. Transesophageal echocardiography is the current standard diagnostic imaging modality in many medical centers. Aortic intramural hematoma is a variant of aortic dissection whose natural history and prognosis have not been well studied. We performed transesophageal echocardiography in patients with aortic intramural hematoma to determine the echocardiographic characteristics and echocardiographic evolution of this lesion, impact on patient management, and patient outcome. METHODS AND RESULTS: Twenty-one consecutive patients with aortic intramural hematoma confirmed anatomically (four patients) or with an additional diagnostic imaging technique (17 patients) underwent a transesophageal echocardiographic examination. Fifteen patients with longstanding hypertension had chest or back pain, and the intramural hematoma was visualized in the ascending aorta (n = 4), along the whole aorta (n = 4), in the descending aorta (n = 6), or in the aortic arch (n = 1). The thickening of the aortic wall was crescentic. Patients with ascending aortic intramural hematoma had the following results: two patients died suddenly, three patients underwent surgery because of increased aortic wall thickening (one patient) or secondary intimal tear (two patients), and the remaining three patients had regression of the hematoma. Patients with hematoma confined to the descending aorta and the patient with aortic arch involvement (n = 7) had a different result: one patient died from aortic rupture and the remaining six patients did well. Six patients had a traumatic aortic injury, and the intramural hematoma was located along the descending thoracic aorta. The thickening of the aortic wall was circular in five patients and crescentic in one. Three of these patients had normalized thickness of the aortic wall on follow-up transesophageal echocardiographic studies. The other three patients died from multiorgan system failure. Aortography showed a reduction of the diameter of the aortic lumen in four patients; diameter in the remaining 17 patients was normal. CONCLUSIONS: Aortic intramural hematoma can be detected and monitored by transesophageal echocardiography but not by aortography. Two types of aortic intramural hematoma can be distinguished: (1) traumatic of good prognosis and (2) nontraumatic, which can be an early stage of the classic aortic dissection, with bad prognosis in cases involving the ascending aorta. PMID- 9327709 TI - Mimics of Ebstein's anomaly. AB - The purpose of this study was to determine the most discriminating clinical and echocardiographic features that are most helpful in correctly identifying Ebstein's anomaly of the tricuspid valve from other causes of tricuspid regurgitation. Ebstein's anomaly is an uncommon malformation of the tricuspid valve with diagnostic echocardiographic features. Other cardiac disorders associated with tricuspid valve regurgitation and predominate right-sided heart chamber enlargement can be misdiagnosed as Ebstein's anomaly. All patients who were referred to our institution between 1982 and 1995 with the diagnosis of Ebstein's anomaly but were found to have other abnormalities of the tricuspid value or right ventricle were identified. Their clinical, echocardiographic, and surgical records were reviewed retrospectively. Twenty-two patients (12 males and 10 females), aged 7 to 68 years (mean 33 years), were referred to our institution with the diagnosis of Ebstein's anomaly but were found to have another abnormality that mimicked clinical and diagnostic features of Ebstein's anomaly. The most common initial symptom was exercise intolerance (13 [59%] patients) followed by atrial arrhythmia (seven [32%] patients). Two patients had cyanosis. Three patients had paroxysmal and six had chronic atrial fibrillation/flutter. Cardiomegaly on chest x-ray film was noted in 18 (82%) patients. Referral diagnosis of Ebstein's anomaly had been made by echocardiography (12 patients), cardiac catheterization (four patients), both techniques (five patients), and echocardiography and magnetic resonance imaging (one patient). All 22 patients had predominate right atrial and right ventricular enlargement, and 18 (82%) of 22 patients also had right ventricular dysfunction. However, Ebstein's anomaly was confidently ruled out with repeat comprehensive echocardiography at our institution by establishing (1) absence of significant apical displacement of the septal tricuspid valve leaflet (> or = 8 mm/m2) and (2) lack of a redundant, elongated, anterior tricuspid valve leaflet in all 22 patients (100%). All had significant tricuspid regurgitation caused by tricuspid valve dysplasia (nine patients), tricuspid valve prolapse (four patients), trauma (four patients), right ventricular dysplasia (three patients), endocarditis (one patient), and annular dilation caused by free pulmonary regurgitation (one patient). In all 15 patients who subsequently underwent surgery (tricuspid valve repair [seven patients] or replacement [eight patients]), the absence of Ebstein's anomaly was confirmed. Echocardiographic absence of the characteristic degree of displacement of the septal leaflet of the tricuspid valve (> or = 8 mm/m2) and the presence of a nonelongated, nonredundant anterior tricuspid valve leaflet consistently excluded the diagnosis of Ebstein's anomaly. Under such circumstances, other anomalies of the tricuspid valve or right ventricle were consistently identified. Recognition of the mimics of Ebstein's anomaly had important surgical implications. PMID- 9327711 TI - Prominent venous valves in hypoplastic right hearts. AB - To investigate the hypothesis that embryologic abnormalities in the venous valves may be associated with abnormal cardiac development, we examined the right atrial morphologic characteristics in 20 hearts with underdevelopment of the right heart and 17 normal hearts. In the study group, 16 (80%) of the patients had significantly enlarged eustachian valves, one (5%) was slightly enlarged, and three (15%) were smaller than expected. Five (25%) had cor triatriatum dexter. In comparison, eustachian valves in the control specimens were prominent in only one (6%), normal in five (29%), and almost absent in eight (47%). The thebesian valve was also more prominent in the study cohort when compared with controls (p < 0.05). No other morphologic features of the right atrium analyzed in this study differed from those found in normal specimens. We speculate that failure of the venous valves to regress appropriately may create abnormalities in fetal circulation that predispose the fetus to maldevelopment of the right heart structures. PMID- 9327710 TI - Renin-angiotensin system: genes to bedside. AB - Atherosclerosis and its vascular sequela are responsible for considerable morbidity and mortality rates. Several risk factors have been implicated in the pathogenesis of atherosclerosis, and the search for other risk factors continues on the medical horizon. Renin-angiotensin system (RAS), a multienzyme, multilocale axis, has been extensively studied as an important mediator of atherosclerosis. Recently, the tissue-based angiotensin system has been suggested as the most significant pathway of RAS. A genetic polymorphism in the human gene for the angiotensin-converting enzyme (ACE), one of the two enzymes of RAS, has been found to have a strong association with higher risk for acute coronary events, sudden cardiac death, vascular restenosis after angioplasty, and idiopathic and hypertrophic cardiomyopathy. Clinical and animal data support angiotensin II to be the final common pathway in the enzyme cascade of RAS and ACE as the key enzyme in the generation of Angiotensin II. ACE gene polymorphism appears to modify expression of cellular and free ACE levels and could represent a genetic marker for cardiovascular disease. PMID- 9327712 TI - Pulmonary hypertension in patients with thalassemia major. AB - To evaluate the pulmonary artery pressure in patients with thalassemia major, Doppler echocardiography was performed in 33 patients with thalassemia major (aged 2 to 24 years) and 33 normal controls. Pulmonary artery pressure was estimated by (1) measuring the systolic transtricuspid gradient from tricuspid regurgitation and adding it to the right atrial pressure, estimated by the response of the inferior vena cava to deep inspiration, and (2) measuring the time to peak velocity of pulmonary flow. The results showed that of 33 patients, 28 had tricuspid regurgitation with a pulmonary systolic pressure ranging from 18 to 94 mm Hg (47 +/- 15 mm Hg). Pulmonary systolic pressure was > 30 mm Hg in all 22 patients > 10 years old and in four of six patients < 10 years old. Pressure correlated with left ventricular ejection fraction (r = -0.74), the ratio of mitral peak early diastolic flow velocity and peak velocity during the atrial contraction (r = 0.52), age (r = 0.56), and total blood transfusion units (r = 0.59). The pulmonary time to peak velocity was shortened compared with controls (p < 0.05). We concluded that pulmonary hypertension, as another cardiovascular complication of multiple factors of cause, seems to occur more frequently and at an early stage of the cardiac involvement in patients with thalassemia major. PMID- 9327713 TI - Coil occlusion of patent ductus arteriosus with detachable coils. AB - Twenty-five patients (mean age 7.0 +/- 4.8 years) underwent transcatheter coil occlusion of patent ductus arteriosus with detachable coils. The minimum diameter of the ductus arteriosus ranged from 1.0 to 4.2 mm (mean 2.6 +/- 0.9 mm). A single-coil technique was used in 17 patients, double- (six patients) or triple coil (two patients) techniques were used in eight patients. The coil was not detached until sufficient shape and position of implanted coils were confirmed. All patients had successful implantation of coils regardless of the morphologic characteristics of the ductus. Immediately after the occlusion, heart murmurs were abolished in all patients. Color-flow mapping showed complete closure in 16 (64%) patients immediately after and 20 (80%) patients 1 month after the procedure. No significant complications occurred. The advantages of this detachable coil system may reduce coil migration and allow safer and more reliable execution of this procedure. PMID- 9327714 TI - Hyperbaric oxygen and thrombolysis in myocardial infarction: the "HOT MI" pilot study. AB - Hyperbaric oxygen treatment (HBO) in combination with thrombolysis has been demonstrated to salvage myocardium in acute myocardial infarction in the animal model. Therefore a randomized pilot trial was undertaken to assess the safety and feasibility of this treatment in human beings. Patients with an acute myocardial infarction (AMI) who received recombinant tissue plasminogen activator (rTPA) were randomized to treatment with HBO combined with rTPA or rTPA alone. Sixty-six patients were included for analysis. Forty-three patients had inferior AMIs (difference not significant) and the remainder had anterior AMIs. The mean creatine phosphokinase level at 12 and 24 hours was reduced in the patients given HBO by approximately 35% (p = 0.03). Time to pain relief and ST segment resolution was shorter in the group given HBO. There were two deaths in the control group and none in those treated with HBO. The ejection fraction on discharge was 52.4% in the group given HBO compared with 47.3% in the control group (difference not significant). Adjunctive treatment with HBO appears to be a feasible and safe treatment for AMI and may result in an attenuated rise in creatine phosphokinase levels and more rapid resolution of pain and ST segment changes. PMID- 9327715 TI - Concentrations of serum interleukin-8 after successful cardiopulmonary resuscitation in patients with cardiopulmonary arrest. AB - To assess differences in serum interleukin-8 concentrations in resuscitated and nonresuscitated patients after cardiopulmonary resuscitation (CPR), and to compare changes of interleukin-8 levels with hemodynamic variables after restoration of spontaneous circulation, 39 patients with out-of-hospital cardiopulmonary arrest who had undergone CPR were studied. Venous blood samples were taken after CPR and 1 and 2 hours after restoration of spontaneous circulation to measure serum interleukin-8 levels by the enzyme-linked immunosorbent assay method. The median serum interleukin-8 levels after CPR were significantly higher in resuscitated than in nonresuscitated patients (64.9 pg/ml; range 30.2 to 1497 vs 0 pg/ml; range 0 to 31.6 pg/ml; p < 0.001). One and 2 hours after restoration of spontaneous circulation, the median serum interleukin-8 levels were 96.2 pg/ml and 155.4 pg/ml, respectively. Mean values of systolic blood pressure immediately after and 1 and 2 hours after restoration of spontaneous circulation were 117 +/- 9 mm Hg, 130 +/- 11 mm Hg, and 136 +/- 13 mm Hg, respectively. No significant correlations were found between hemodynamic values and serum interleukin-8 levels. In conclusion, successful initial resuscitation was associated with increased serum interleukin-8 concentrations. The interleukin-8 levels remained at high values 2 hours after restoration of spontaneous circulation. PMID- 9327717 TI - Gemfibrozil treatment of hypertriglyceridemia: improvement on fibrinolysis without change of insulin resistance. AB - The fibrinolytic and metabolic changes associated with gemfibrozil treatment of hypertriglyceridemia were evaluated in 16 patients with type IV hyperlipidemia by criteria of triglyceride levels > 250 mg/dl and total cholesterol levels < 220 mg/dl. The plasma triglyceride level was significantly lower (323 +/- 71 vs 189 +/- 57 mg/dl; p = 0.000) and high-density lipoprotein cholesterol level significantly higher (33.5 +/- 4.6 vs 38.0 +/- 6.7 mg/dl; p = 0.005) after 3 to 4 months of gemfibrozil treatment. However, the glucose and insulin metabolism measured by oral glucose challenge and insulin suppression tests showed no significant changes after gemfibrozil therapy. In contrast, plasma plasminogen activator inhibitor-1 antigen (36.9 +/- 12.4 vs 27.3 +/- 11.4 ng/ml; p = 0.008) and activity (15.5 +/- 5.5 vs 11.8 +/- 3.0 IU/ml; p = 0.009) and tissue plasminogen activator antigen (13.2 +/- 4.0 vs 10.4 +/- 3.7 ng/ml; p = 0.007) were significantly depressed, and tissue plasimogen activator activity (0.57 +/- 0.31 vs 0.69 +/- 0.38 IU/ml; p = 0.015) was significantly elevated by gemfibrozil. The data indicate that lowering plasma triglyceride and raising high density lipoprotein cholesterol levels by gemfibrozil treatment also improved the fibrinolytic system without changes of insulin resistance and glucose intolerance in patients with isolated hypertriglyceridemia. PMID- 9327716 TI - Long-term effects of angiotensin-converting enzyme inhibitors and calcium antagonists on the right and left ventricles in essential hypertension. AB - To compare the effects of chronic antihypertensive treatment on left and right ventricular structure and function, 24 patients with mild to moderate, never treated hypertension were randomized to receive fosinopril (20 mg daily) or amlodipine (10 mg daily) for 12 months. At baseline and subsequently at the end of third, sixth, and twelfth months, each patient underwent an integrated echocardiographic study and noninvasive ambulatory blood pressure monitoring. Both drugs significantly reduced blood pressure, casual or monitored (p < 0.01), and left ventricular mass index (from 125 +/- 32 to 100 +/- 12 gm/m2 [p < 0.02] with amlodipine and from 106 +/- 18 to 89 +/- 10 gm/m2 [p < 0.02] with fosinopril). The decrease in left ventricular mass was essentially caused by a reduction of ventricular thickness. Free right ventricular wall thickness was also lowered in both groups, more consistently with amlodipine (from 8.0 +/- 2.1 to 6.4 +/- 0.8 mm; p < 0.01), without an increase in plasma natriuretic peptide and insulin concentrations or heart rate. With both treatments, the decrease in ventricular mass was not associated with impairment of systolic function, whereas a trend toward an improvement of Doppler echocardiographic indexes of biventricular diastolic function was observed. In conclusion, both amlodipine and fosinopril induced similar qualitative effects on anatomy and function of both ventricles. The clinical meaning of these observations must be defined further by means of adequately sized prospective trials. PMID- 9327718 TI - Elevated plasma lipid hydroperoxides in patients with coronary artery disease. AB - Sustained presence of lipid peroxides in the circulation and their plasma carrier is a controversial issue. Particularly, there is no firm evidence for an increased plasma lipid peroxide level in patients with atherosclerosis. In this study, a strong correlation was found between plasma total lipid hydroperoxide and lipid hydroperoxide content of LDL cholesterol (r = 0.882; p < 0.001; n = 16). Lipid hydroperoxides in plasma were carried almost exclusively (89%) in LDL. In 70 patients tested 3 months after coronary artery bypass graft surgery with a specific assay, plasma lipid hydroperoxide levels were significantly increased when compared with matched healthy controls (4.31 +/- 0.23 nmol/ml and 2.34 +/- 0.13 nmol/ml, p < 0.0001, patients vs controls, respectively). These concentrations are 10 times lower than those detected by the nonspecific thiobarbituric acid assay. However, considering the in vitro concentration range in which oxidized LDL exerts important atherogenic effects, the elevated plasma lipid hydroperoxide levels measured in atherosclerotic patients have pathologic significance. PMID- 9327719 TI - Molecular and cellular prospects for repair, augmentation, and replacement of the failing heart. AB - Molecular and cellular biology offer the promise of new approaches to the treatment of heart failure. This article discusses the basic science background, the current state of investigation, and the potential for therapeutic application of these new sciences. It also emphasizes the limitations and unknowns in this frontier. Three approaches are presented: First, increasing the number of myocytes in the heart, previously held to be untenable because postnatal cardiomyocytes do not divide, may be possible by regulating the cell cycle to reinduce cardiac growth. Also, nonmyocytes extant in the heart may be coaxed into differentiating into cardiomyocytes, or exogenous muscle cells may be grafted into the myocardium. Second, cardiac function may be augmented by molecular therapies that increase contractile protein function or regulate beta-adrenergic receptors or Ca++ channels. Third, improved prospects for transplantation of the failed heart may occur by genetic modification of a xenograft donor heart that reduces the chance of immune rejection by the human recipient. The formulation for the successful application of any of these therapies depends on not only the creativity of scientists but also the wisdom of physicians. PMID- 9327720 TI - Inhibition of NF-kappa B activity induces apoptosis in murine hepatocytes. AB - Recently we have demonstrated that inhibition of the nuclear factor (NF)-kappa B/Rel family of transcription factors induces apoptosis of B cells. Interestingly, mice lacking the relA gene encoding the p65 subunit of NF-kappa B exhibit embryonic lethality at days 15 to 16 of gestation, accompanied by massive destruction of liver via apoptosis. To determine whether p65 protein plays a direct role in hepatocyte survival, we employed a nontransformed murine hepatocyte (NMH) cell line, which maintains to a high degree the differentiated hepatocyte phenotype. Exponentially growing NMH cells were found to possess a constitutive level of functional classical (p50/p65) NF-kappa B as assayed by electrophoretic mobility shift analysis, antibody supershift, and transient transfection assays. Treatment of NMH cells with the proteasome inhibitor lactacystin, which prevents degradation of the NF-kappa B inhibitor proteins I kappa B, induced apoptosis. Direct inhibition of the endogenous NF-kappa B activity by microinjection of NMH cells with purified specific inhibitor I kappa B-alpha-glutathione-S-transferase fusion protein or an antibody against p65 protein induced apoptosis. These findings suggest that expression of NF-kappa B/Rel activity in murine hepatocytes acts directly to promote survival of these cells and suggest that apoptosis observed in hepatocytes of mice lacking relA is a direct effect of p65 deficiency. PMID- 9327722 TI - Bile ductular damage induced by methylene dianiline inhibits oval cell activation. AB - Administration of 2-acetylaminofluorene (2-AAF) given before a two-thirds partial hepatectomy (PHx), results in suppression of hepatocyte proliferation and stimulation of oval cell proliferation. Our objective in this study was to examine the oval cell response and associated alpha-fetoprotein (AFP) gene expression by combining 2-AAF with selective hepatic damage caused by either carbon tetrachloride (CCl4) exposure or by PHx. We also examined oval cell response with the above two protocols (2-AAF/CCl4 and 2-AAF/PHx) as affected by previous bile ductular damage caused by 4,4'-methylene dianiline (4,4' diaminodiphenylmethane, DAPM) exposure. DAPM is an aromatic diamine, known to cause bile ductular damage in both humans and animals. Using the protocols of 2 AAF/ CCl4 and 2-AAF/PHx, when DAPM was given 24 hours before the hepatic injury, no oval cell proliferation was seen (histological) and AFP expression was not detected by Northern blot analysis. These results provide direct evidence that oval cells are closely associated with the biliary epithelial cells and supports the theory that hepatic oval cells may originate from cells derived from either intraportal or periportal ductules. PMID- 9327721 TI - Evidence for intrathecal synthesis of alternative pathway complement activation proteins in experimental meningitis. AB - Complement has been shown to contribute to intrathecal inflammation in bacterial meningitis. However, the cellular source of complement in the infected central nervous system has not been determined. In this study, we analyzed protein and mRNA expression of two alternative pathway complement activation proteins, C3 and factor B, in the brains of mice with Listeria monocytogenes meningitis. Complement protein levels were found elevated in the cerebrospinal fluid of infected mice, compared with mock-infected animals. In the course of the disease, enhanced C3 and factor B mRNA expression was detected on pyramidal neurons and Purkinje cells within 6 hours, peaking at 12 hours and then gradually decreasing by 72 hours after infection. In addition, leukocytes infiltrating the subarachnoid space, within 12 to 24 hours, expressed mRNA for C3 and factor B. The cellular infiltration increased dramatically up to 72 hours. Intraperitoneal injection of tumor necrosis factor (TNF)-alpha up-regulated C3 and factor B mRNA expression on neurons in normal mice, suggesting that TNF-alpha may represent one cytokine regulating complement expression in this model of bacterial meningitis. However, additional mediators may be involved in regulation of intrathecal complement expression, as infected mice deficient of TNF/lymphotoxin-alpha genes did not demonstrate attenuated complement expression in the brain. PMID- 9327723 TI - Amyloid A protein amyloidosis induced in apolipoprotein-E-deficient mice. AB - Apolipoprotein E (apoE) is a constituent of lipoproteins other than low-density lipoprotein, and it principally acts in the transport and metabolism of plasma cholesterol and triglyceride. ApoE is a minor constituent of various kinds of amyloidoses and may play a role as a pathological chaperone for fibrillogenesis of amyloid fibril protein with the amyloid P component and proteoglycans. In this study, we examined the role of apoE in amyloidogenesis in vivo in apoE-deficient mutant mice with amyloid A protein (AA) amyloidosis induced by inflammatory stimulation. Amyloid deposition was seen in six of nine C57BL/6J control mice and in six of eight apoE-deficient mutant mice after the intraperitoneal and subcutaneous injections of the mixture of complete Freund's adjuvant and Mycobacterium butyricum. Moreover, amyloid deposition in apoE-deficient mice as well as C57BL/6J control mice started 48 or 72 hours after injection of amyloid enhancing factor and silver nitrate, although the amount of amyloid deposit in C57BL/6J control mice was slightly larger than that in apoE-deficient mice. These amyloid deposits reacted with anti-mouse AA antibody were seen in the perifollicular area of the spleen. Immunoreactivity of apoE was seen irregularly in the amyloid deposits of C57BL/6J control mice but not in the amyloid deposit of apoE-deficient mice. From these results, we concluded that apoE is not always necessary for amyloid deposition and that the existence of apoE might slightly accelerate AA amyloid deposition in the earliest phase of AA amyloid deposition. PMID- 9327725 TI - Feline atopic dermatitis. A model for Langerhans cell participation in disease pathogenesis. AB - Atopic dermatitis is a disorder characterized by cutaneous exanthemata as a consequence of exaggerated eczematous reactions to topical and systemic allergens. Langerhans cells, expressing CD1a and HLA-DR, and dermal dendritic cells, expressing HLA-DR, are known to be potent antigen-presenting cells and are thought to play an important role in the pathogenesis of atopic dermatitis. The immunophenotype of lesional skin in atopic dermatitis in humans involves increased numbers of CD1a+/MHC class II+ dendritic cells in addition to activated T cells, mast cells, and macrophages. To establish feline skin as a model for the study of human atopic dermatitis, and to elucidate the role of dendritic cells in feline atopic dermatitis, we investigated the presence of CD1a+ cells and MHC class II+ cells in the epidermis and dermis of lesional feline skin and in skin of healthy control animals. Immunohistochemistry revealed that MHC class II+ epidermal dendritic cells were CD1a+ in normal feline skin and significantly increased numbers of CD1a+ cells and MHC class II+ cells were present in the epidermis and dermis of lesional skin. These data provide the first correlative documentation of CD1a expression by feline dendritic cells containing Birbeck granules, and indicate the utility of feline skin in the study of human cutaneous atopy. PMID- 9327724 TI - Inducible nitric oxide synthase expression in coronary arteries of transplanted human hearts with accelerated graft arteriosclerosis. AB - Inducible nitric oxide synthase (iNOS) is a high-output isoform of NOS that produces nitric oxide (NO), a nonspecific immune effector molecule. In some animal models of autoimmunity, the induction of iNOS has been shown to lead to inflammation and tissue damage, and it has been suggested that iNOS is an immune mediator in humans as well. Using in situ hybridization and immunohistochemical techniques, we demonstrate that iNOS mRNA and protein are present in the coronary arteries of transplanted human hearts with accelerated graft arteriosclerosis (AGA). iNOS is expressed in cells morphologically consistent with macrophages in the neointima of 7 of 10 of the transplanted vessels with AGA that were examined. In serial sections, these same cells express the macrophage marker CD68. In contrast, iNOS is absent from five native coronary arteries with atherosclerosis and absent from two normal coronary arteries. Although iNOS is expressed in macrophages in AGA, its role in the pathogenesis of AGA is unknown. PMID- 9327726 TI - Increased oxidative DNA damage and hepatocyte overexpression of specific cytochrome P450 isoforms in hepatitis of mice infected with Helicobacter hepaticus. AB - A recently discovered bacterium, Helicobacter hepaticus, infects the intrahepatic bile canaliculi of mice, causing a severe chronic hepatitis culminating in liver cancer. Thus, it affords an animal model for study of bacteria-associated tumorigenesis including H. pylori-related gastric cancer. Reactive oxygen species are often postulated to contribute to this process. We now report that hepatitis of male mice infected with H. hepaticus show significant increases in the oxidatively damaged DNA deoxynucleoside 8-hydroxydeoxyguanosine, with the degree of damage increasing with progression of the disease. Perfusion of infected livers with nitro blue tetrazolium revealed that superoxide was produced in the cytoplasm of hepatocytes, especially in association with plasmacytic infiltrates near portal triads. Contrary to expectations, Kupffer cells, macrophages, and neutrophils were rarely involved. However, levels of cytochrome P450 (CYP) isoforms 1A2 and 2A5 in hepatocytes appeared to be greatly increased, as indicated by the number of cells positive in immunohistochemistry and the intensity of staining in many cells, concomitant with severe hepatitis. The CYP2A5 immunohistochemical staining co-localized with formazan deposits resulting from nitro blue tetrazolium reduction and occurred in nuclei as well as cytoplasm. These findings suggest that CYP2A5 contributes to the superoxide production and 8-hydroxydeoxyguanosine formation, although reactive oxygen species from an unknown source in the hepatocytes leading to CYP2A5 induction or coincidental occurrence of these events are also possibilities. Three glutathione S-transferase isoforms, mGSTP1-1 (pi), mGSTA1-1 (YaYa), and mGSTA4-4, also showed striking increases evidencing major oxidative stress in these livers. PMID- 9327727 TI - Can K-ras codon 12 mutations be used to distinguish benign bile duct proliferations from metastases in the liver? A molecular analysis of 101 liver lesions from 93 patients. AB - It can be difficult to distinguish benign bile duct proliferations (BDPs) from well-differentiated metastatic peripancreatic adenocarcinomas on histological grounds alone. Most peripancreatic carcinomas harbor activating point mutations in codon 12 of the K-ras oncogene, suggesting that K-ras mutational status may provide a molecular basis for distinguishing BDPs from liver metastases. The ability of tests for mutations in codon 12 of K-ras to make this distinction was examined in a two-part study. In the first part we determined the K-ras mutational status of 56 liver lesions and 48 primary peripancreatic adenocarcinomas obtained from 48 patients. In the second part of this study an additional 45 liver lesions were studied. In the first 48 patients, activating point mutations in codon 12 of K-ras were detected in 28 (61%) of the 46 primary carcinomas, in 8 (100%) of 8 liver metastases, in 2 (6.5%) of 31 BDPs, and in none (0%) of 14 liver granulomas. Three BDPs and two primary carcinomas did not amplify. To further estimate the prevalence of K-ras mutations in BDPs we analyzed an additional series of 45 mostly incidental BDPs for K-ras mutations. Three (6.7%) of these 45 harbored K-ras mutations. These results suggest that K ras mutations may be useful in distinguishing BDPs from metastases in the liver; however, there is some overlap in the mutational spectra of BDPs and pancreatic carcinomas. PMID- 9327728 TI - Allelic deletion and mutation of the von Hippel-Lindau (VHL) tumor suppressor gene in pancreatic microcystic adenomas. AB - An association between pancreatic microcystic (serous) adenomas (MCAs) and von Hippel-Lindau (VHL) disease has been suggested. However, genetic alterations of the VHL gene in MCAs of the pancreas have never been reported. In this study, we performed genetic analysis of 12 pancreatic MCAs. In 2 cases, VHL disease was documented clinically, and 10 cases were sporadic. For LOH analysis, tumor and normal pancreatic cells were procured from formalin-fixed, paraffin-embedded material using tissue microdissection. After DNA extraction, the samples were amplified by polymerase chain reaction using the polymorphic markers D3S2452, D3S1110, D3S192, and D3S656. In addition, the sporadic tumors were analyzed for VHL gene mutations using probes 3b/10b and K55/K56. Both MCAs associated with VHL disease showed LOH with at least one of the microsatellite markers tested. Among the 10 sporadic cases, 7 tumors showed LOH at the VHL gene locus. A somatic VHL gene mutation on exon 2 was documented in one sporadic case. The study provides the first direct genetic evidence for the role of the VHL gene in MCA tumorigenesis. Furthermore, VHL gene alterations may be detected in both VHL associated and sporadic pancreatic MCAs. PMID- 9327729 TI - Correlation between Bcl-2 expression and histopathology in diethylnitrosamine induced mouse hepatocellular tumors. AB - It is generally accepted that suppression of apoptosis in chemically initiated hepatocytes results in promotion of rodent hepatocarcinogenesis. Using immuno histochemical methods, I studied the expression of Bcl-2, an anti-apoptotic protein, in hepatocellular tumors of B6C3F1 mice. Although normal mouse hepatocytes did not express detectable amounts of Bcl-2, most diethylnitrosamine induced tumors were positive for this protein. Virtually all of the Bcl-2 positive tumors were composed of small basophilic hepatocytes, whereas the rare cases of Bcl-2-negative tumors demonstrated an eosinophilic appearance. To confirm this difference, tumors initiated with diethylnitrosamine and promoted by phenobarbital were also studied, as this initiation-promotion protocol has been shown to selectively produce eosinophilic lesions. All such tumors were immunohistochemically negative for Bcl-2. The relatively infrequent basophilic tumors found with phenobarbital treatment, however, did express Bcl-2. Thus, the concordance with basophilia was observed regardless of the nature of the promotion agent. These results indicate that the two types of tumors are qualitatively distinct and may develop through independent mechanisms. PMID- 9327730 TI - Selective deposits of versican in the extracellular matrix of restenotic lesions from human peripheral arteries. AB - Although a large percentage of the volume of human restenotic arterial lesions is occupied by extracellular matrix (ECM), the composition and organization of this ECM are not well characterized. In this study, restenotic segments taken from 30 human peripheral arteries by directional atherectomy at times ranging from 13 days to 36 months after angioplasty were analyzed for specific patterns of ECM composition and organization by light and electron microscopic histochemistry and immunohistochemistry. Histochemical analysis revealed the presence of distinct zones, enriched either in proteoglycans or fibrillar collagen. Most sections contained these regions juxtaposed to each other. The frequency of these two distinct ECMs did not change as a function of time after angioplasty. The collagen-rich zone usually contained elongated smooth muscle cells spaced close together except in regions resembling fibrous plaques. The proteoglycan-rich ECM contained both elongated and stellate-shaped smooth muscle cells randomly arranged and separated by wide distances. This region resembled the loose connective-tissue-containing myxoid region typical of restenotic lesions. Immunohistochemical analysis of these regions revealed that the proteoglycan containing zone stained intensely for versican, a large interstitial chondroitin sulfate proteoglycan, whereas the collagen-containing areas were mostly negative for versican but positive for type I collagen. The versican-positive regions also immunostained for biglycan, a small leucine-rich dermatan sulfate proteoglycan, and sparsely for elastin. However, both of these ECM molecules were present in the versican-negative type I collagen-positive regions of the lesions. These results suggest that the development of restenotic lesions involves localized deposits of specific ECM molecules that may play a role in the asymmetric renarrowing of this tissue after angioplasty. PMID- 9327731 TI - Expression and function of endothelial cell alpha v integrin receptors in wound induced human angiogenesis in human skin/SCID mice chimeras. AB - Accumulating evidence indicates that endothelial cell integrins that bind to the matrix proteins associated with inflammation and wound healing are involved in the process of angiogenesis. The integrins containing the alpha v subunit appear to be particularly important. To study the involvement of these receptors in human angiogenesis, a model of wound-associated human angiogenesis was established in human skin transplanted onto severe combined immunodeficient (SCID) mice. Using this model, we studied the expression of several alpha v integrins and tested the hypothesis that blockage of the alpha v beta 3 integrin would inhibit human angiogenesis during human wound healing. These studies revealed that the alpha v beta 3, alpha v beta 5, and alpha v beta 6 integrins are up-regulated briefly during wound angiogenesis with different patterns of expression and that inhibition of the alpha v beta 3 integrin blocked new vessel formation during human wound healing. PMID- 9327732 TI - Dedifferentiation of atrial cardiomyocytes as a result of chronic atrial fibrillation. AB - Chronic atrial fibrillation was induced in goats by electrical pacing. After 9 to 23 weeks of sustained atrial fibrillation, the morphology of the atrial structures was examined. The majority of the cardiomyocytes exhibited marked changes in their cellular substructures, with the replacement of sarcomeres by glycogen as the main characteristic. Using immuno-histochemical staining procedures, we assessed the expression and organization of contractile and cytoskeletal proteins in these cases and compared them with the expression and organization of these proteins in normal atria. Part of the atrial cardiomyocytes acquired a dedifferentiated phenotype, as deduced from the re-expression of alpha smooth muscle actin, the disappearance of cardiotin, and the staining patterns of titin, which resembled those of embryonic cardiomyocytes. From these results we conclude that chronic atrial fibrillation induces myocardial dedifferentiation. This model of chronic atrial fibrillation in goats offers the possibility to study the time course of changes in cardiac structure during sustained atrial fibrillation and after cardioversion. PMID- 9327733 TI - The potential role of BAX and BCL-2 expression in diffuse alveolar damage. AB - Apoptosis of type II pneumocytes has been identified in diffuse alveolar damage (DAD), is associated with p53 and WAF1 expression, and may be of pathogenetic importance. BAX, a homologue of BCL-2, is induced by p53 and is a promoter of apoptosis. The proapoptotic effect of BAX is negatively regulated by its binding with BCL-2. In this study, we sought to investigate that role of BAX and BCL-2 in DAD. We hypothesized that alterations in BAX and BCL-2 expression may be important in determining the susceptibility of type II pneumocytes and interstitial cells to apoptosis. Twenty-eight cases of DAD and 16 control cases (i.e., lung tissues adjacent to resected tumors) were retrieved from the files of the University of Utah and the Armed Forces Institute of Pathology. Immunohistochemical stains were performed with antigen retrieval by microwave using antibodies recognizing BAX and BCL-2. The percentage of positively staining pneumocytes and interstitial cells was estimated in each case to the nearest 10%. BAX expression was markedly increased in pneumocytes and interstitial cells in DAD compared with control lung tissues. In DAD, BAX was identified on an average of 80% of alveolar pneumocytes (range 30 to 100%) and 70% of interstitial cells (range 20 to 90%). In control lungs, BAX was identified on an average of 10% of pneumocytes (range 0 to 20%) but not in interstitial cells. Focal BCL-2 staining was identified in interstitial myofibroblasts in 7 of 25 cases of DAD but was only identified in bronchiolar epithelium of control lungs. These results suggest that the induction of BAX in DAD may enhance the susceptibility of alveolar epithelial cells to apoptosis, whereas BCL-2 expression may contribute to the absence of apoptosis in interstitial myofibroblasts. Expression of BCL-2 in interstitial myofibroblasts may contribute to the development of pulmonary fibrosis in some patients. PMID- 9327734 TI - Respiratory infection in lipid-fed rabbits enhances sudanophilia and the expression of VCAM-1. AB - The pathogenesis of atherosclerosis has been related to infection of the arterial wall, but it is not clear whether this occurs before or after the development of lipid-containing lesions. Respiratory bacterial infection increases the expression of vascular cell adhesion molecule-1 (VCAM-1). We therefore examined whether a similar infection would enhance atherosclerosis in New Zealand White rabbits fed chow supplemented by 15% (w/w) egg yolk for 50 days. Rabbits with naturally acquired respiratory infection by Pasteurella multocida, pathogen-free (SPF) animals infected by P. multocida in the laboratory, and age-matched SPF rabbits maintained in a disease-free environment were used. Endothelial cells expressing VCAM-1 in the aorta between intercostal arteries 3 and 5 were identified using anti-VCAM-1 (Rb1/9) and an alkaline-phosphatase-linked secondary antibody and quantified in Hautchen preparations. The remainder of the aorta was stained with Sudan IV to show lipid deposition. The expression of VCAM-1 (mean +/ SEM per 10,000 cells) was 22 +/- 8 (n = 5) in the lipid-fed SPF rabbits, significantly different from that in the lipid-fed rabbits with naturally occurring infection (190 +/- 51 (n = 5)) or from rabbits infected in the laboratory (106 +/- 25 (n = 5)). The extent of Sudanophilia was significantly greater in the naturally infected rabbits (8.3 +/- 1.2%) or infected SPF rabbits (10.3 +/- 1.8%) than in the SPF rabbits (2.7 +/- 0.8%; P < 0.05). Antibiotic treatment in naturally infected rabbits reduced the number of cells expressing VCAM-1 and the extent of the Sudanophilia to baseline levels. Thus, Sudanophilia is enhanced by bacterial infection in rabbits fed egg yolk and is associated with a significant increase in VCAM-1. PMID- 9327736 TI - Detection of numerical and structural alterations and fusion of chromosomes 16 and 1 in low-grade papillary breast carcinoma by fluorescence in situ hybridization. AB - Intracystic papillary breast tumors, including intraductal papilloma and low grade intracystic papillary carcinoma, constitute a group for which differential diagnosis is frequently difficult. We examined the status of chromosomes 16 and 1 by multicolor fluorescence in situ hybridization (FISH) analyses and the DNA ploidy patterns by flow cytometry in 26 intracystic papillary tumors. Alterations of chromosomes 16 and 1 were detected by FISH in 93 and 85%, respectively, of 14 low-grade papillary carcinomas, and the latter alterations always concurred with the former. Two-color FISH using probes for the D1Z1 (1q12) and D16Z2 (16cen) loci or the D1Z1 and D16Z3 (16q11) loci showed that fusion of chromosomes 16 and 1, mostly with breakpoints distal to 16q11.2 and proximal to 1q12, occurred in 77% of the papillary carcinomas. DNA aneuploidy was detected in 6% of these carcinomas. No papilloma showed these chromosome alterations or DNA aneuploidy. Chromosome 16 and 1 fusions appeared to occur frequently in diploid breast carcinomas and to be involved in the acquisition of a malignant phenotype by duct epithelial cells. We suggest that two-color FISH methods for detecting 1;16 fusions might be applicable as supportive methods for the differential diagnosis of intracystic papillary breast tumors. PMID- 9327735 TI - Pulmonary hemodynamics modify the rat pulmonary artery response to injury. A neointimal model of pulmonary hypertension. AB - Hemodynamic factors have profound influences on blood vessels. To test the hypothesis that hemodynamic conditions modify the pattern of remodeling in response to injury, monocrotaline (MCT) injury in Sprague-Dawley rats was followed 1 week later by left pneumonectomy to increase blood flow to the right lung. Right pulmonary artery remodeling in these MCT plus pneumonectomy animals was compared with animals receiving MCT or pneumonectomy alone. Neointimal changes developed in more than 90% of all right lung intra-acinar vessels 5 weeks after MCT injury (4 weeks after pneumonectomy). Neointimal lesions did not develop in untreated animals or in animals receiving MCT or pneumonectomy only. Animals with a neointimal pattern of remodeling developed severe right ventricular hypertrophy (RVH) whereas animals with a medial hypertrophy pattern of remodeling (MCT only) developed moderate RVH compared with control animals. Neointimal lesions and RVH were similar whether injury preceded pneumonectomy or vice versa. To exclude the possibility that neointimal lesions resulted from injury plus post-pneumonectomy compensatory lung growth, rather than injury plus increased flow, a left subclavian-pulmonary artery anastomosis was substituted for pneumonectomy. Neointimal lesions and severe RVH developed in these animals but were not seen in animals receiving either MCT or anastomosis only. These studies demonstrate an important role for hemodynamics in determining the pattern of pulmonary vascular remodeling after injury. PMID- 9327737 TI - CXCR-4 (Fusin), a co-receptor for the type 1 human immunodeficiency virus (HIV 1), is expressed in the human brain in a variety of cell types, including microglia and neurons. AB - Entry of the type 1 human immunodeficiency virus into most cells requires the presence of the CD4 protein in combination with one of several recently described co-receptors. CXCR-4 (fusin) was the first identified, and it serves as co receptor for T-cell-line tropic (T-tropic) HIV-1 isolates. To determine the expression of CXCR-4 in the brain, a major target of HIV pathology, we used immunohistochemistry and reverse transcriptase polymerase chain reaction with CXCR-4-specific antibodies and probes. We found that CXCR-4 was expressed in several cell types in brain, but notably in neurons and microglia, a finding that was replicated in tissue culture. The study of the expression of CXCR-4 in the brain, which may be one of many chemokine receptors in the central nervous system, may provide further insight into the interactions between brain cells, pathogens, and the immune system, and help understand the pathogenesis of HIV dementia. PMID- 9327740 TI - HOXC4, HOXC5, and HOXC6 expression in primary cutaneous lymphoid lesions. High expression of HOXC5 in anaplastic large-cell lymphomas. AB - Homeobox (HOX) genes are involved in the lineage-specific differentiation of bone marrow stem cells. Recently, we reported a largely similar expression pattern of HOXC4 and HOXC6 in normal and neoplastic cells of the lymphoid lineage. In contrast, HOXC5 was specifically expressed in a subset of B-cell non-Hodgkin's lymphomas (B-NHL) but not in normal lymphocytes or lymphoid leukemias. This might suggest a role for HOXC5 in the pathogenesis of these lymphomas. In the present study the expression of HOXC4, HOXC5, and HOXC6 in primary cutaneous lymphomas was investigated. Using RNA in situ hybridization (RISH) and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), we found strong expression of HOXC4 and HOXC6 in all, except one, primary cutaneous lymphomas and all reactive cutaneous lymphoid infiltrates. Interestingly, a strong expression of HOXC5 in primary anaplastic CD30+ large T-cell lymphomas was found. RISH was consistently negative for HOXC5 in all other types of primary cutaneous B- and T cell lymphomas. However, by semiquantitative RT-PCR these lymphomas showed a weak expression of HOXC5 mRNA. Therefore, we concluded that these lymphomas express low constitutive levels of HOXC5 mRNA. Furthermore, HOXC5 expression was consistently absent in reactive cutaneous lymphoid infiltrates, hyperplastic tonsils and lymph nodes, and peripheral blood lymphocytes either unstimulated or stimulated by a cocktail of CD3 and CD28 antibodies. As a strong expression of HOXC5 in primary cutaneous lymphomas was observed only in anaplastic large T-cell lymphomas and reactive control tissues lacked HOXC5 expression, these data strongly support a role for HOXC5 in the genesis of anaplastic large-T-cell lymphomas. PMID- 9327738 TI - Involvement of transcription factor encoded by the mouse mi locus (MITF) in apoptosis of cultured mast cells induced by removal of interleukin-3. AB - Mast cells develop when spleen cells of mice are cultured in the medium containing interleukin (IL)-3. Cultured mast cells (CMCs) show apoptosis when they are incubated in the medium without IL-3. We obtained CMCs from tg/tg mice that did not express the transcription factor encoded by the mi gene (MITF) due to the integration of a transgene at its 5' flanking region. MITF is a member of the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of transcription factors. We investigated the effect of MITF on the apoptosis of CMCs after removal of IL-3. When cDNA encoding normal MITF ((+)-MITF) was introduced into tg/tg CMCs with the retroviral vector, the apoptosis of tg/tg CMCs was significantly accelerated. The mutant mi allele represents a deletion of an arginine at the basic domain of MITF. The apoptosis of tg/tg CMCs was not accelerated by the introduction of cDNA encoding mi-MITF. The overexpression of (+)-MITF was not prerequisite to the acceleration of the apoptosis, as the apoptotic process proceeded faster in +/+ CMCs than in mi/mi CMCs. The Ba/F3 lymphoid cell line is also dependent on IL-3, and Ba/F3 cells show apoptosis after removal of IL-3. The c-myc gene encodes another transcription factor of the bHLH-Zip family, and the overexpression of the c-myc gene accelerated the apoptosis of Ba/F3 cells. However, the overexpression of (+)-MITF did not accelerate the apoptosis of Ba/F3 cells. The (+)-MITF appeared to play some roles for the acceleration of the apoptosis specifically in the mast cell lineage. PMID- 9327741 TI - Postnatal lung function and morphology in transgenic mice expressing transforming growth factor-alpha. AB - Developmental changes in lung morphology and physiology during postnatal alveolarization were assessed in transgenic mice expressing transforming growth factor-alpha (TGF-alpha) in pulmonary type II cells under control of the surfactant protein C gene promoter. TGF-alpha transcripts were identified in respiratory epithelial cells at 1 day of age to adulthood. Enlargement of alveolar airspaces and fibrosis were detected as early as 1 week of age, and the increased airspace progressed with advancing age. Specific lung compliance was significantly increased in lungs of transgenic mice by 2 weeks of age and was associated with airflow obstruction. Chronic expression of TGF-alpha in the lungs of newborn transgenic mice caused remodeling of the developing lung during the period of postnatal alveolarization, resulting in markedly enlarged parenchymal airspace, pulmonary fibrosis, and physiological abnormalities including airway obstruction and increased lung compliance. PMID- 9327739 TI - Expression of cytokine mRNA in lentivirus-induced arthritis. AB - Infection of goats with the lentivirus caprine arthritis encephalitis virus (CAEV) leads to persistent infection and development of chronic arthritis. We analyzed the expression of cytokines and viral RNA in the joints of goats at early time points after experimental infection with CAEV and in those of animals suffering from chronic arthritis as a result of natural infection. In situ hybridization experiments showed that the pattern of cytokine expression in caprine arthritis was similar to that found in rheumatoid arthritis (RA), with a few cells expressing the lymphocyte-derived cytokines interferon (IFN)-gamma and interleukin (IL)-2 and rather more cells expressing monocyte chemoattractant protein (MCP)-1, IL-6, and tumor necrosis factor (TNF)-alpha. IFN-gamma mRNA expression in experimentally infected joints peaked at day 12 and was mostly detected in areas containing viral RNA. At later time points, no IFN-gamma- or virus-expressing cells were found in inflamed joints but both were again detected in goats with severe arthritis. Interestingly, at the clinical stage of arthritis reflecting the chronic stage of infection, the inflammatory lesion was found to be immunologically compartmentalized. Humoral immune responses and cell-mediated immune responses appeared to concurrently occur in distinct areas of the synovial membrane. PMID- 9327743 TI - Loss of type IV collagen alpha 5 and alpha 6 chains in human invasive prostate carcinomas. AB - Type IV collagen, a major component of basement membranes, is organized in a network responsible for the mechanical resistance of the basement membranes. It also plays a key role in epithelial cell adhesion to basement membranes. This study was designed to investigate the distribution of type IV collagen alpha chains in normal, preneoplastic, and malignant prostate basement membranes. For this purpose, immunohistochemistry using specific antibodies raised against the different alpha-chains of type IV collagen was performed in eight normal samples, six prostatic intraepithelial neoplasia, and 20 malignant lesions of the prostate. Our results demonstrate the presence of the "novel" alpha 5 (IV) and alpha 6 (IV) chains along with the "classical" alpha 1 (IV)/alpha 2 (IV) chains in the basement membrane of the normal prostate gland. The alpha 3 (IV) chain was never detected in any prostate specimen. Prostatic intraepithelial neoplasia showed a similar immunostaining pattern to that found in normal glands. In cancer gland basement membranes, we demonstrate for the first time a specific loss of the alpha 5 (IV) and alpha 6 (IV) chains, whereas the classical alpha 1 (IV) and alpha 2 (IV) chains were consistently exhibited. Additionally, type VII collagen colocalized with alpha 5 (IV) collagen chain, and these two proteins, which were always observed in normal and prostatic intraepithelial neoplasia gland basement membranes, were lost in invasive carcinoma basement membranes. This observation raises questions about the possible association or cooperation between alpha 5 (IV)/alpha 6 (IV) chains and anchoring fibrils in prostate glands basement membrane. PMID- 9327742 TI - Activation of the NF-kappa B and I kappa B system in smooth muscle cells after rat arterial injury. Induction of vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1. AB - The NF-kappa B transcription factor family and its inhibitory proteins (I kappa B) form an autoregulatory system that has been linked to endothelial gene expression and vascular disease. To determine the role of the NF-kappa B/I kappa B system in smooth muscle cells (SMCs) in vivo, the present study used the balloon catheter injury model in the rat carotid artery. The NF-kappa B family members p50, p65, p52, c-Rel, and RelB as well as the inhibitor proteins I kappa B alpha, I kappa B beta, and p105 were present in uninjured arteries as determined by immunoblotting. Using electromobility shift assays, low levels of constitutively activated p50, p65, and c-Rel were seen in normal carotid arteries and a fivefold induction occurred during times of rapid SMC proliferation and neointima formation after balloon denudation. Furthermore, immediately after injury, the levels of I kappa B alpha, I kappa B beta, and p105 were dramatically reduced. Expression of the NF-kappa B-regulated genes, vascular cell adhesion molecule (VCAM)-1 and monocyte chemotactic protein (MCP)-1, was apparent in SMCs within 4 hours after injury. Macrophage infiltration occurred in parallel with the expression of VCAM-1 and MCP-1, and these inflammatory cells were present on the luminal surface of injured vessels during intimal lesion formation. In chronically denuded vessels, the SMCs on the luminal surface continued to express high levels of VCAM-1 and MCP-1, which may account for the increased presence of macrophages. Together, these findings link the activation of NF-kappa B to intimal lesion formation and to the inflammatory response associated with SMCs after vascular injury. PMID- 9327744 TI - Expression of interleukin-8 by human melanoma cells up-regulates MMP-2 activity and increases tumor growth and metastasis. AB - Expression of interleukin-8 (IL-8) by human melanoma cells correlates with their metastatic potential. Moreover, UV-B irradiation of primary cutaneous melanoma cells induces IL-8 mRNA and protein production and increases both tumor growth and metastasis in nude mice. Although IL-8 has been shown to be an angiogenic factor, the biological consequences of increased IL-8 production by melanoma cells and the role of IL-8 in the metastatic process remains unclear. The purpose of this study was to determine the role of IL-8 in tumor growth and metastasis of human melanoma cells. Nonmetastatic SB-2 melanoma cells with negligible levels of IL-8 were transfected with IL-8 cDNA and subsequently analyzed for changes in their tumorigenic and metastatic potential. Enforced expression of IL-8 rendered the melanoma cells highly tumorigenic and increased their metastatic potential as compared with parental and control transfected cells. The IL-8-transfected cells displayed up-regulation in M(r) 72,000 collagenase type IV (MMP-2) mRNA and collagenase activity and increased invasiveness through Matrigel-coated filters. Moreover, when the MMP-2 promoter was linked upstream of the chloramphenicol acetyltransferase (CAT) reporter gene, CAT activity was up-regulated in IL-8 but not in control transfected cells, suggesting that IL-8 is involved in MMP-2 gene transcription. Activation of type IV collagenase by IL-8 can enhance the invasion of host stroma by the tumor cells and increase angiogenesis and, hence, metastasis. PMID- 9327745 TI - Role of glycosaminoglycans in determining the helicity of paired helical filaments. AB - It is known from previous work that tau is the main component of paired helical filaments (PHFs) and that it can assemble in vitro into polymers resembling PHFs when high concentrations of protein are used. In the search for molecules that can facilitate tau polymerization, a component of neurofibrillary tangles, heparan sulfate (or its more sulfated form, heparin), and other glycosaminoglycans have been tested. Glycosaminoglycans, in the sulfated but not in the unsulfated form, facilitate not only tau assembly but also the formation of polymers resembling PHFs. Conversely, PHFs were found to contain heparan sulfate and chondroitin sulfate. Heparinase or chondroitinase treatment of PHFs results in the formation of straight structures. All of these results suggest a role for sulfated glycosaminoglycans in determining the helicity of PHFs. PMID- 9327747 TI - Diabetes induces changes in glomerular development and laminin-beta 2 (s-laminin) expression. AB - Offspring of diabetic mothers have developmental renal abnormalities; thus, we investigated the effects of the diabetic milieu on kidney development. Four groups of host rats, including insulin-deficient and insulin-treated streptozotocin-induced diabetic rats, normal controls, and insulin-treated nondiabetic rats, were prepared. After 38 days, rats received ocular implants of E14 fetal rat kidneys. Nine days later the fetal kidney grafts were harvested for analysis of glomerular development and expression of fibronectin, laminin, laminin-beta 2, and alpha-smooth muscle actin and m170, two additional markers of mesangial maturation. The rate of glomerular maturation was delayed in grafts placed in hyperglycemic, insulin-deficient diabetic rats. These glomeruli contained few mesangial cells or matrix, and laminin-beta 2 expression was reduced as compared with controls. Mesangial expression of alpha-smooth muscle actin and m170 was not detected. In contrast, grafts placed in insulin-treated diabetic animals had increased numbers of mesangial cells and expanded mesangial matrix. The content of laminin-beta 2 and expression of m170 and alpha-smooth muscle actin were also increased in these grafts. These data show that hyperglycemia and insulin status influence laminin isoform expression and play important roles in mesangial development. PMID- 9327746 TI - Classical Hodgkin's disease. Clinical impact of the immunophenotype. AB - Antibodies against CD15, -30, and -20 are often used to support morphological diagnosis of Hodgkin's Disease (HD). The classical HD, i.e., the non-lymphocyte predominance types, are CD15+, CD30+, and CD20- in general. However, the results for CD15 are less clear-cut in many studies, showing up to 40% of classical HD that lack positivity for this maker. Little is currently known about the relevance of antigen expression in relation to clinical outcome in HD. Therefore, the three markers were analyzed in 1751 cases from the German Hodgkin Study Group, using micro-wave epitope retrieval to optimize staining sensitivity. Eighty-three percent of the cases showed a classical immunophenotype (CD15+, CD30+, CD20-), twelve percent lacked CD15 positivity (CD15-, CD30+, CD20-), and five percent showed other combinations. For 1286 cases, clinical follow-up was available, which revealed significant differences for freedom from treatment failure (P = 0.0022) and overall survival (P = 0.0001) between cases with classical immunophenotype and CD15 negativity (CD30+, CD20-). Multivariate Cox regression using the three markers, age, sex, histology, stage, B-symptoms (fever, sweats, weight loss > 10% of body weight), hemoglobin, and erythrocyte sedimentation rate as factors showed that lack of CD15 expression in classical HD is an independent negative prognostic factor for relapses (P = 0.022) and survival (P = 0.0035). In conclusion, immunohistochemistry is able to identify classical HD cases with unfavorable clinical outcome. PMID- 9327748 TI - Podoplanin, novel 43-kd membrane protein of glomerular epithelial cells, is down regulated in puromycin nephrosis. AB - Puromycin aminonucleoside nephrosis (PAN), a rat model of human minimal change nephropathy, is characterized by extensive flattening of glomerular epithelial cell (podocyte) foot processes and by severe proteinuria. For comparison of expression of glomerular membrane proteins of normal and PAN rats, a membrane protein fraction of isolated rat glomeruli was prepared and monoclonal antibodies were raised against it. An IgG-secreting clone designated LF3 was selected that specifically immunolabeled podocytes of normal but not of PAN rats. The antigen of LF3 IgG was identified as a 43-kd glycoprotein. Molecular cloning of its cDNA was performed in a delta gt11 expression library prepared from mRNA of isolated rat glomeruli. The predicted amino acid sequence indicated a 166-amino-acid integral membrane protein with a single membrane-spanning domain, two potential phosphorylation sites in its short cytoplasmic tail, and six potential O glycosylation sites in the large ectodomain. High amino acid sequence identities were found to membrane glycoproteins of rat lung and bone and mouse thymus epithelial cells as well as to a phorbol-ester-induced protein in a mouse osteoblast cell line and to a canine influenza C virus receptor. In PAN, expression of this 43-kd protein was selectively reduced to < 30%, as determined by quantitative immunogold electron microscopy, immunoblotting, and Northern blotting. These data provide evidence that transcription of the 43-kd transmembrane podocyte glycoprotein is specifically down-regulated in PAN. To indicate that this protein could be associated with transformation of arborized foot processes to flat feet (Latin, pes planus) we have called it podoplanin. PMID- 9327749 TI - Comparative genomic hybridization of malignant fibrous histiocytoma reveals a novel prognostic marker. AB - DNA sequence copy number changes were studied by comparative genomic hybridization (CGH) along all chromosomes in 58 samples of malignant fibrous histiocytoma (MFH). The material consisted of 43 primary tumors (9 of myxoid and 34 of storiform-pleomorphic subtype), 13 local recurrences (2 myxoid and 11 storiform-pleomorphic), and 2 metastases (1 myxoid and 1 storiform-pleomorphic). Genetic aberrations, with a mean of 5.5 changes per sample (range, 0 to 22), were detected in 47 of 58 samples (81%). The minimal common regions of the most frequent gains were 1p31 (33%), 9q31 (29%), 5p14-pter (26%), 7q32 (24%), and 7p15 pter (22%). High-level amplifications were detected in 16 of the 58 samples (28%). High-level amplification of 13q31-qter was seen in four tumors (7%); other high-level amplifications were more sporadic. Losses of DNA sequences were less frequent than gains. The minimal common regions of the most common losses were 13q21 (21%) and 13q22 (21%). Statistically significant correlation was found between gain of 7q32 and the rates of worse metastasis-free survival (P = 0.01) and overall survival (P = 0.004). The gain of 7q32 retained its prognostic significance also in a multivariate analysis with tumor size and grade. Gain of 1p31 was associated with a trend to decreased overall survival. Gains of 5p14 pter and 9q31 and losses of 13q21 and/or 13q22 did not have any prognostic value; neither did the total number of aberrations, total number of gains, or total number of losses per sample. PMID- 9327751 TI - Use of FISH analysis for prostate tumors and other tissue types. PMID- 9327752 TI - The efficacy of in situ PCR, CARD and nanogold systems for gene detection. PMID- 9327750 TI - Dephosphorylation of endotoxin by alkaline phosphatase in vivo. AB - Natural substrates for alkaline phosphatase (AP) are at present not identified despite extensive investigations. Difficulties in imagining a possible physiological function involve its extremely high pH optimum for the usual exogenous substrates and its localization as an ecto-enzyme. As endotoxin is a substance that contains phosphate groups and is usually present in the extracellular space, we studied whether AP is able to dephosphorylate this bacterial product at physiological pH levels. We tested this in intestinal cryostat sections using histochemical methods with endotoxin from Escherichia coli and Salmonella minnesota R595 as substrate. Results show that dephosphorylation of both preparations occurs at pH 7.5 by AP activity. As phosphate residues in the lipid A moiety determine the toxicity of the molecule, we examined the effect of the AP inhibitor levamisole in vivo using a septicemia model in the rat. The results show that inhibition of endogenous AP by levamisole significantly reduces survival of rats intraperitoneally injected with E. coli bacteria, whereas this drug does not influence survival of rats receiving a sublethal dose of the gram-positive bacteria Staphylococcus aureus. In view of the endotoxin-dephosphorylating properties of AP demonstrated in vitro, we propose a crucial role for this enzyme in host defense. The effects of levamisole during gram-negative bacterial infections and the localization of AP as an ecto enzyme in most organs as well as the induction of enzyme activity during inflammatory reactions and cholestasis is in accordance with such a protective role. PMID- 9327753 TI - Is HIV found in the cytoplasm of dendritic cells? PMID- 9327754 TI - Bacterial toxins block endothelial wound repair. Evidence that Rho GTPases control cytoskeletal rearrangements in migrating endothelial cells. AB - We investigated the effect of bacterial toxins that modify and inactivate Rho GTP binding proteins on the migratory response of endothelial cells to wounding. C3 transferase from Clostridium botulinum, EDIN from Staphylococcus aureus, and toxin A from Clostridium difficile blocked migration of human umbilical vein endothelial cells (HUVECs) in an in vitro wound repair assay. Migrating HUVECs expressed actin microspikes (maximum at 10 minutes after wounding), ruffles (maximum at 12 hours), and fibers (maximum at 24 hours), and within these actin structures, vinculin-containing focal complexes/adhesions were formed. C3 Transferase ADP ribosylated RhoA, RhoB, and RhoC in HUVECs and abolished the formation of actin stress fibers/focal adhesions but had no effect on expression of microspikes, ruffles, or the associated vinculin-containing focal complexes. Similar results were obtained with EDIN and toxin A. These results indicate that endothelial cells migrating into a wounded area express distinct combinations of actin/vinculin structures in a spatially and temporally coordinated manner. The GTPase Rho selectively controls the formation of actin fibers/focal adhesions that occurs 2 to 24 hours after wounding. A mechanism is proposed by which Rho specific bacterial toxins could influence vascular repair, angiogenesis, or atherosclerosis. PMID- 9327755 TI - Role of free apolipoprotein A-I in cholesterol efflux. Formation of pre-alpha migrating high-density lipoprotein particles. AB - This article characterizes products formed by the interaction of purified apolipoprotein (apo) A-I and human fibroblasts. Fibroblasts were incubated with different concentrations of purified apoA-I (1 to 30 micrograms/mL) in tissue culture medium for different periods of time (0 to 24 hours). The medium was then characterized by one- (agarose) and two-dimensional (agarose: polyacrylamide nondenaturing gradient gel) electrophoresis. At any given concentration of apoA I, the rate of cellular cholesterol efflux appeared linear over 24 hours. Incubating purified apoA-I with fibroblasts for 4 hours, we detected five pre alpha lipoproteins with particle sizes between 114 and 684 kDa. Formation of pre alpha lipoproteins was concentration-dependent. At low concentrations (below 5 micrograms/mL apoA-I), all purified apoA-I (with pre-beta mobility) was converted to pre-alpha lipoproteins. At higher concentrations (greater than 5 micrograms/mL apoA-I), more apoA-I remained with pre-beta mobility. The pre-alpha lipoproteins were characterized by colocalization of apoA-I particles with 14C-cholesterol and 32P-phospholipids. Results showed that the pre-alpha particle of lowest molecular weight contained phospholipid and apoA-I but no cholesterol. The remaining pre alpha particles contained all three substances. When pre-alpha particles were subjected to ultracentrifugation, all particles floated at d < 1.21 g/mL with some of the smallest phospholipid apoA-I only particles being present in the d > 1.21 g/mL fraction. Based on these results, we postulated that in the first stages of reverse cholesterol transport, pre-alpha lipoproteins are formed by the interaction of lipid free apoA-I and peripheral cells. PMID- 9327756 TI - Evidence that apolipoprotein A-IMilano has reduced capacity, compared with wild type apolipoprotein A-I, to recruit membrane cholesterol. AB - Human carriers of apolipoprotein (apo) A-IMilano are heterozygous for an Arg173- >Cys substitution in the apoA-I primary sequence; despite severe reductions in HDL cholesterol concentrations, affected individuals do not develop coronary heart disease, suggesting that apoA-IMilano may possess antiatherogenic properties. As the beneficial effects of wild-type apoA-I are linked to its role in HDL cholesterol transport, we examined the capacity of apoA-IMilano to recruit cell cholesterol and activate lecithin:cholesterol acyltransferase (LCAT) (two key events in the antiatherogenic reverse cholesterol transport pathway). ApoA IMilano and wild-type apoA-I were expressed in Chinese hamster ovary cells, and their ability to recruit membrane phospholipid and cholesterol for the assembly of nascent HDL was compared. Both clonal cell lines exhibited similar levels of apolipoprotein accumulation in serum-free medium (approximately 2 micrograms/mg cell protein per 24 hours), and 15% of each apolipoprotein was associated with membrane lipids to form nascent HDL (d = 1.063 to 1.21 g/mL). SDS-PAGE showed that a majority (66 +/- 12%) of the lipidated apoA-IMilano was in the homodimer form. Compositional analyses revealed that apoA-IMilano nascent HDL had a significantly lower (P < .001) unesterified cholesterol/phospholipid mole ratio (0.47 +/- 0.10) than wild-type apoA-I complexes (1.29 +/- 0.14), indicating that apoA-IMilano had a reduced capacity to recruit cell cholesterol. In addition to the reduced unesterified cholesterol/phospholipid ratio, apoA-IMilano nascent HDL consisted mostly of small 7.4-nm particles compared with wild-type apoA-I, in which 11- and 9-nm particles predominated. Despite these changes in nascent HDL particle size and composition, apoA-IMilano activated LCAT normally. We conclude that, even though apoA-IMilano is a normal activator of LCAT, it is less efficient that wild-type apoA-I in recruiting cell cholesterol, suggesting that the putative antiatherogenic properties attributed to apoA-IMilano may be unrelated to the initial stages of reverse cholesterol transport. PMID- 9327757 TI - Induction of cyclooxygenase-2 in human saphenous vein and internal mammary artery. AB - Within vessels, cyclooxygenase (COX) is expressed constitutively (COX-1) in endothelial cells where its production of prostacyclin is thought to contribute to the maintenance of vascular integrity. Recently, a novel isoform of COX, COX 2, has been described that is induced in animal arterial vessels after physical damage or exposure to proinflammatory cytokines. However, induction of COX-2 in human vessels has not been characterized. Moreover, the relative ability of arteries and veins to express COX-2 has not been addressed. Thus, we have compared the ability of segments of human saphenous vein and internal mammary artery, obtained from the same patient, to express COX-2 activity and mRNA after organ culture in the presence and absence of interleukin-1 beta. COX-2 metabolites, measured by radioimmunoassay, were released by both the internal mammary artery and saphenous vein in the following rank order: prostaglandin E2 > or = prostacyclin thromboxane A2. Inclusion of interleukin-1 beta in the culture medium increased the release of prostanoids by the saphenous vein but not by the internal mammary artery. However, the selective COX-2 inhibitor NS-398 significantly attenuated prostacyclin release from both tissues. Northern blot analysis showed no detectable COX-2 mRNA in freshly prepared saphenous vein or internal mammary artery. In contrast, after 48 hours in organ culture, low levels of COX-2 mRNA were detected in both internal mammary artery and saphenous vein, an effect that was greatly increased by interleukin-1 beta. These observations show that COX-2 is induced in human saphenous vein and internal mammary artery and suggest that this may occur in humans after coronary artery bypass graft surgery. The induction of COX-2 and subsequent release of prostacyclin may represent an endogenous defense mechanism against endothelial damage incurred during surgical preparation of these vessels for bypass. PMID- 9327758 TI - Repeated balloon injury of rat aorta. A model of neointima with attenuated inhibition by heparin. AB - Repeated arterial injury, because it challenges already activated cells, may elicit a reaction that differs from that provoked by a single injury. We compared the response of rat aorta to single and double balloon injury and tested the inhibitory effect of heparin in both situations. For repeated injury, the first and second lesions were induced 3 weeks apart. Two weeks after repeated injury, the neointima that existed from the first lesion had expanded, with significant increases in intima-media wet weight and its DNA and elastin content and in the intima-to-media (I/M) thickness ratio. Two days after repeated injury, the expression of proliferating cell nuclear antigen (PCNA) was enhanced in both the media and the intima, indicating that cells from both layers are involved in the aortic response to a second lesion. As established previously, treatment with heparin (continuous intravenous administration, 50 IU/kg.h-1) almost totally suppressed the response to single injury. However, heparin only attenuated the response to repeated injury, with a partial decrease in intima-media wet weight and its DNA and elastin content and in I/M thickness ratio. PCNA labeling showed that heparin inhibited the proliferative activity in medial cells much more strongly than in intimal cells. In conclusion, repeated aortic injury elicits a reaction of both the media and preexisting neointima. In this mixed response, neointimal smooth muscle cells are less sensitive than medial cells to inhibition by heparin, which results in a weakened effect of the drug on the fibromuscular reaction. PMID- 9327759 TI - Plasma lipoprotein (a) levels in men and women consuming diets enriched in saturated, cis-, or trans-monounsaturated fatty acids. AB - Studies that have shown adverse effects of trans-unsaturated fatty acids on plasma lipoprotein (a) [Lp(a)] levels have used levels of trans-fatty acid that are higher than those in the average U.S. diet. This study was conducted to clarify the effects on Lp(a) of trans-fatty acids levels commonly found in U.S. diets. Lp(a) levels were measured in a double-blind study of 29 men and 29 women who ate 4 controlled diets in random order for 6 weeks each. Fatty acids represented 39% to 40% of energy. The diets were: (1) Oleic (16.7% of energy as oleic acid); (2) Moderate trans (3.8% of energy as trans-monoenes, approximately the trans content of the U.S. diet); (3) High trans (6.6% of energy as trans monoenes); (4) Saturated (16.2% of energy as lauric plus myristic plus palmitic acids). The Saturated diet lowered Lp(a) levels significantly (by 8% to 11%). Compared to the Oleic diet, the trans diets had no adverse effect on Lp(a) levels when all subjects were considered collectively. A subset with initially high levels of Lp(a) (> or = 30 mg/dL), however, responded to the High trans diet with a slight (5%) increase in Lp(a) levels relative to the Oleic and Moderate trans diets. Thus, in amounts commonly found in the typical U.S. diet, saturated fatty acids consistently decrease Lp(a) concentrations. The adverse effects of replacing cis- with trans-fatty acids are only suggestive and are restricted to high trans intakes in subjects with high Lp(a) levels. PMID- 9327760 TI - A common mutation in the methylenetetrahydrofolate reductase gene (C677T) increases the risk for deep-vein thrombosis in patients with mutant factor V (factor V:Q506). AB - Hyperhomocysteinemia is a frequent risk factor for deep-vein thrombosis. A common mutation (C677T) in the gene encoding for methylenetetrahydrofolate reductase (MTHFR) is responsible, in the homozygous state, for decreased enzyme activity and mild hyperhomocysteinemia and is associated with increased risk for cardiovascular disease. We studied the prevalence of C677T MTHFR in 77 patients with deep-vein thrombosis and in 154 age- and sex-matched healthy control subjects. In the same individuals, we also evaluated the frequency of the coexistence of C677T MTHFR with mutant factor V:Q506, a common risk factor for deep-vein thrombosis. Sixteen patients (20.8%) and 35 control subjects (22.7%) were homozygous for the C677T MTHFR mutation (odds ratio [OR] = 0.8, 95% confidence interval [CI] = 0.4-2.0). Sixteen patients (20.8%) and 4 control subjects (2.6%) had factor V:Q506; of them, 10 patients and 3 control subjects had isolated factor V:Q506 (adjusted OR = 6.3, 95% CI = 1.6-25.3) and 6 patients and 1 control subject also had C677T MTHFR (adjusted OR = 17.3, 95% CI = 2.0 152.9). The OR for the coexistence of the two mutations was 65% to 75% higher than the expected joint effect calculated by either an additive (OR = 6.0) or multiplicative (OR = 4.4) model. The homozygous C677T mutation of MTHFR per se is not a risk factor for deep-vein thrombosis but increases the risk associated with factor V:Q506. Due to the high prevalence of C677T MTHFR, it is likely that previous studies, which did not look for this mutation, overestimated the relative risk of thrombosis associated with factor V:Q506 alone. PMID- 9327761 TI - High density lipoproteins stimulate mitogen-activated protein kinases in human skin fibroblasts. AB - Protein kinase C (PKC) seems to play an important role in many of HDL effects on cells, including removal of excess cholesterol. HDL removes cholesterol by at least two mechanisms. One mechanism involves desorption/diffusion of cholesterol from the plasma membrane onto the acceptor particle, whereas the second is mediated by apolipoproteins and may involve intracellular translocation of cholesterol to the plasma membrane for subsequent efflux. In this report, we examined the possibility that mitogen-activated protein (MAP) kinase is one of the downstream events from HDL activation of PKC. Using a gel kinase assay with myelin basic protein incorporated into the gel, HDL (50 micrograms protein/mL) stimulated multiple kinases of 42, 50, 52, 58, and 60 kDa. The 42-kDa protein kinase, corresponding to the unresolved MAP kinases ERK1 and ERK2 based on immunoblotting, was activated over 2-fold by HDL. HDL activated all identified kinases in a concentration- and time-dependent manner, which became maximal within 5 to 10 minutes and remained activated for at least 60 minutes. HDL activation of MAP kinase seems to be partially mediated by PKC, because down regulation of PKC and known PKC inhibitors inhibited the HDL effect by 40 to 50%. Free apolipoproteins A-I (10 micrograms/mL) and A-II (10 micrograms/mL) had no significant effect on MAP kinase activation. Moreover, modifying HDL with trypsin or tetranitromethane, which abolishes apolipoprotein-mediated cholesterol efflux, had no effect on HDL activation of MAP kinase. These results suggest that HDL activates MAP kinase via multiple signal transduction pathways that are likely involved in an HDL effect unrelated to apolipoprotein-mediated cholesterol translocation and efflux. PMID- 9327762 TI - Interactions between lifestyle-related factors and the ApoE polymorphism on plasma lipids and apolipoproteins. The EARS Study. European Atherosclerosis Research Study. AB - To elucidate how the apolipoprotein (apo) E polymorphism and modifiable factors interact in explaining plasma lipid and apolipoprotein levels, we studied 1448 young adults (18 to 26 years old), participating in the European Atherosclerosis Research Study (EARS). Venous blood was collected after an overnight fast. Modifiable factors, eg, body mass index (BMI), waist-to-hip ratio (WHR), tobacco and alcohol consumption, and physical activity, were determined by using standardized protocols. Associations of modifiable factors with apoE levels were homogeneous across apoE phenotypes. In contrast, correlations of BMI with total cholesterol and apoB levels, as well as correlations between WHR and apoB, were significantly (P < .05 to P < .01) stronger in E2 carriers than in subjects with other phenotypes. Total cholesterol and apoB levels were comparable in E2 carriers in the upper tertile of BMI or WHR to those in E3/3 subjects, suggesting that the lowering effect of the E2 allele was no longer present. The inverse association between the plasma cholesteryl linoleate-to-oleate ratio, a marker for the dietary polyunsaturated-to-saturated fatty acid ratio, and triglycerides was also stronger in E2 carriers (-0.33 versus -0.17 in E3/3 and -0.24 in E4 carriers). Associations with other modifiable factors were notably consistent across apoE phenotypes. Gender and modifiable factors explained three times more (31%) of the interindividual variation in apoB levels in E2 carriers than in E3/3 subjects (9%) or E4 carriers (14%), mainly due to a larger variance explained by BMI. Our results suggest that the apoE polymorphism acts in a relatively uniform manner, independently of lifestyle. However, the associations of adiposity to total cholesterol and apoB levels appear to be stronger in apoE2 carriers. PMID- 9327763 TI - Effects of short-term exercise on female platelet function during different phases of the menstrual cycle. AB - Previous studies have shown that premenopausal women have a low incidence of cardiovascular diseases, and that acute exercise affects male platelet function in an intensity-dependent manner. To investigate whether acute exercise affects female platelet function differently from males, sixteen sedentary women in the midfollicular phase or midluteal phase received strenuous or moderate exercise on a bicycle ergometer. Before and immediately after exercise, platelet adhesiveness, adenosine diphosphate-induced platelet aggregation and intracellular calcium concentration elevation, platelet cAMP and cGMP contents, urinary 11-dehydro-TXB2 and 6-keto-prostaglandin F1 alpha levels, and plasma nitric oxide metabolite level were determined. Our results showed no differences in exercise performance and in resting platelet function between two menstrual phases, with little change in urinary eicosanoid metabolites and platelet cAMP levels under all experimental conditions. In addition, for women in the midfollicular phase, (1) strenuous exercise increased platelet adhesiveness, adenosine-diphosphate-induced platelet aggregation, and intracellular calcium concentration elevation, whereas moderate exercise suppressed them; (2) moderate exercise enhanced plasma nitric oxide metabolite and platelet cGMP levels. In contrast, none of these platelet functions was affected by acute exercise in the midluteal phase. Therefore, we conclude that acute exercise affects female platelet function in an intensity-dependent manner in the midfollicular phase but not in the midluteal phase. The irresponsiveness of platelets to acute exercise in the luteal phase may partially explain why premenopausal women have a lower incidence of cardiovascular diseases than men. PMID- 9327764 TI - Alu-repeat polymorphism in the gene coding for tissue-type plasminogen activator (t-PA) and risks of myocardial infarction among middle-aged men. AB - An Alu-repeat polymorphism in the gene coding for tissue-type plasminogen activator has been described recently, and it has been hypothesized that this polymorphism may predict risk of coronary thrombosis. In a prospective cohort of nearly 15,000 apparently healthy men, presence of an Alu-repeat insertion/deletion (I/D) polymorphism in the gene coding for tissue-type plasminogen activator was determined among 369 study participants who subsequently suffered a first myocardial infarction (cases) and among a group of 369 age- and smoking-matched study participants who remained free of reported cardiovascular disease during follow-up (controls). The distributions of the II, DI, and DD genotypes of the tissue-type plasminogen activator polymorphism among men who subsequently suffered myocardial infarction (0.30, 0.50, 0.21) were virtually identical to those who remained free of disease (0.29, 0.50, 0.21; P = .9). There was no evidence of association between the Alu insertion polymorphism and risks of future myocardial infarction in models assuming either allelic recessive (relative risk, 1.05; 95% confidence interval, 0.8 to 1.4, P = .8) or allelic dominant (relative risk, 1.04; 95% confidence interval, 0.7 to 1.5, P = .8) modes of inheritance, nor were associations found in analyses stratified by age, family history, hypercholesterolemia, or the presence of other risk factors for premature coronary disease. Multivariate analysis had no important effects on these relationships. In this cohort of middle-aged US men, the presence of the insertion allele of the Alu-repeat polymorphism of the tissue-type plasminogen activator gene is not associated with future risks of myocardial infarction. PMID- 9327765 TI - Effect of 17 beta-estradiol on metabolism of acetylated low-density lipoprotein by THP-1 macrophages in culture. AB - Evidence from numerous epidemiological and animal studies has shown a protective effect of estrogens on the development of atherosclerosis. Since not all of the beneficial effects of estrogen can be explained by alterations in plasma lipoprotein profiles, estrogens may have a direct effect on the arterial wall on one or more of the key steps in the pathogenesis of atherosclerosis. In the present study we tested the hypothesis that estrogens decrease macrophage foam cell formation by reducing lipoprotein uptake via the scavenger receptor pathway. Incubation of the human THP-1 macrophage cell line with 17 beta-estradiol reduced the uptake and metabolism of 125 I-labeled human acetylated LDL (acLDL) in a concentration-dependent manner (from 10(-9) to 10(-5) mol/L) by 30% to 40% at the highest concentrations used. This decrease was accompanied by a reduction in cholesterol accumulation and esterification. When chloroquine was used to block lysosomal degradation, 17 beta-estradiol retained its ability to decrease accumulation of acLDL. This finding suggested that the effect of estrogen occurs before degradation of acLDL by lysosomes. 17 beta-Estradiol had no effect on binding of 125I-acLDL at 4 degrees C. When 125I-acLDL was bound at 4 degrees C and warmed to 37 degrees C, less acLDL was internalized and degraded in cells treated with 17 beta-estradiol, due to greater dissociation of the bound acLDL from the surface of estrogen-treated cells during internalization. We conclude that as a result of the estrogen-induced increase in dissociation of acLDL, less lipoprotein cholesterol is delivered to macrophages, resulting in a reduced rate of foam cell formation. This may be one mechanism by which estrogens reduce the development of atherosclerosis. PMID- 9327766 TI - Genetic variation in factor VII associated with variation in plasma lipoprotein(a) concentration. AB - Cross-sectional and prospective studies have shown that individuals with high plasma lipoprotein(a) [Lp(a)] concentrations are at increased risk for coronary heart disease. Size polymorphism of the apolipoprotein(a) [apo(a)] glycoprotein accounts for approximately 35% of the variation in plasma Lp(a) concentrations. However, there is no convincing evidence for associations between plasma Lp(a) and common genetic variation outside APO(a), the gene that encodes apo(a). We tested for association of common genetic variation of candidate genes in lipid metabolism and also of F7 with variation of plasma Lp(a) concentrations in Alberta Hutterites. Variation at codon 353 of F7 has been associated with variation in the plasma factor VII activity (FVIIc), with the 353Q allele associated with lower FVIIc and the 353R allele associated with higher FVIIc. We found significant associations between variation in plasma concentrations of Lp(a) and both apo(a) isoform size and F7 codon 353 genotype (both P < .0001). The effects on plasma Lp(a) concentration of the alleles at codon 353 were additive. The average effects of the F7 353Q and 353R alleles were, respectively, to decrease by 1.71 micrograms/mL and to increase by 0.301 microgram/mL plasma Lp(a) concentration from the sample mean. This suggests that common genomic variation in F7 is associated with variation in plasma Lp(a) concentration. PMID- 9327767 TI - Uptake of type III hypertriglyceridemic VLDL by macrophages is enhanced by oxidation, especially after remnant formation. AB - We previously showed that hypertriglyceridemic VLDL (HTG-VLDL, Sf 60 to 400) from subjects with type III (E2/E2) hyperlipoproteinemia do not induce appreciable cholesteryl ester (CE) accumulation in cultured macrophages (J774A.1). In the present study, we examined whether oxidation of type III HTG-VLDL would enhance their uptake by J774A.1 cells. Type III HTG-VLDL were oxidized as measured by both conjugated-diene formation and increased electrophoretic mobility on agarose gels. Both LDL and type III HTG-VLDL undergo oxidation, albeit under different kinetic parameters. From the conjugated-diene curve, type III HTG-VLDL, compared with LDL, were found to have a 6-fold longer lag time, to take 6-fold longer to reach maximal diene production, and to produce a 2-fold greater amount of dienes but at half the rate (all P < .005). Incubation of macrophages with either native type III HTG-VLDL or LDL (50 micrograms lipoprotein cholesterol/mL media for 16 hours) caused small increases (4-fold and 2.7-fold, respectively) in cellular CE levels relative to control cells (both P = .0001). After 24 hours of CuSO4 exposure, we found that oxidized type III HTG-VLDL and LDL caused a 9.4-fold and 10.5-fold increase, respectively, in cellular CE levels (P = .0001). We next examined whether extending the exposure period for type III HTG-VLDL to CuSO4 beyond 24 hours would further enhance its ability to induce macrophage CE accumulation. After 48 hours of CuSO4 exposure, type III HTG-VLDL and LDL caused 21.3-fold and 11.6-fold increases, respectively, in cellular CE levels (P = .0001). The cellular CE loading achieved with 48 hour-oxidized type III HTG-VLDL was significantly higher than either 24 hour-oxidized type III HTG-VLDL (2.3 fold, P = .003) or 48 hour-oxidized LDL (1.8-fold, P = .012). There was no significant difference between the CE loading achieved by incubation of cells with either 24 hour-oxidized type III HTG-VLDL, 24 hour-oxidized LDL, or 48 hour oxidized LDL (P > or = .518). In this study, we also examined whether partial lipolysis (19% to 50% triglyceride hydrolysis) of type III HTG-VLDL to produce remnants would increase the susceptibility of the lipoprotein to oxidative modification and subsequent cellular CE loading. Forty-eight hour-oxidized type III VLDL-remnants stimulated CE accumulation 30.4-fold over baseline (P = .0001). In contrast, nonoxidized type III VLDL-remnants caused the same very low level of CE loading as did native type III HTG-VLDL (P = .680). The increase in cellular CE levels achieved with 48 hour-oxidized type III VLDL-remnants was significantly higher than that achieved with 48 hour-oxidized type III HTG-VLDL (P = .047). In conclusion, we have shown that oxidized type III HTG-VLDL will induce macrophage CE accumulation well above levels achieved with oxidized LDL. In addition, we also showed that by forming a VLDL-remnant before oxidative modification, we can further enhance macrophage CE accumulation. These results provide a potential mechanism for the atherogenicity of type III HTG-VLDL and their remnants. PMID- 9327768 TI - Lipid transfer inhibitor protein activity deficiency in normolipidemic uremic patients on continuous ambulatory peritoneal dialysis. AB - We previously demonstrated that lipid transfer inhibitor protein (LTIP) is a potent modifier of lipid transfer protein (LTP) function in vitro. Based on these studies, we proposed that LTIP activity is an important determinant of lipoprotein size and composition, which leads to a stimulation of reverse cholesterol transport. To further evaluate this hypothesis, we have studied a normolipidemic, uremic patient population undergoing continuous ambulatory peritoneal dialysis (CAPD) that is deficient in LTIP activity (< 18% of control). LDL from CAPD plasma was triglyceride enriched; the diameters of both CAPD LDL and HDL were increased and CAPD HDL was dominated by the largest subfraction, HDL2b. In CAPD patients, the plasma cholesterol esterification rate was only 61% of control; this decrease was due mainly to the poor reactivity of CAPD lipoproteins. CAPD lipoprotein-deficient plasma promoted twofold greater transfer of radiolabeled cholesteryl ester (CE) between standard lipoproteins than control, although LTP itself was increased only 39%. This twofold increase was not equally expressed among individual lipoprotein classes; CE transfers involving LDL were increased 2.4-fold, whereas those not involving LDL were increased only 50%. In whole plasma, CE net mass transfer to VLDL was slightly increased in CAPD plasma; relative to their CE content, control HDL contributed twofold more CE mass to VLDL than control LDL, but in CAPD plasma this preferential transfer of CE from HDL was absent. Collectively, the aberrations in CAPD lipoprotein composition and metabolism are consistent with the hypothesized role of LTIP. The data further support the role of LTIP in modulating the participation of HDL in CE mass transfers to VLDL. This is the first report of LTIP activity deficiency in humans. PMID- 9327769 TI - HDL deficiency in genetically engineered mice requires elevated LDL to accelerate atherogenesis. AB - In humans, a low HDL concentration is one of the strongest indicators of increased risk for coronary heart disease. Apolipoprotein A-I (apo A-I) synthetic defects results in extremely low HDL levels and are frequently although not invariably associated with premature atherosclerosis. To investigate atherosclerosis susceptibility associated with HDL deficiency alone and in combination with other risk factors, such as high levels of LDL, we have quantified diet-induced atherogenesis in a series of genetically engineered mice, including mice with low HDL levels due to targeted disruption of both apo A-I alleles (AI KO mice), mice with high LDL levels due to expression of a human apolipoprotein B transgene (Btg mice), and mice with combined high LDL and low HDL levels due to the presence of the human apo B transgene and apo A-I knockout alleles, respectively (AI KO/Btg mice). After exposure to an atherogenic diet, AI KO and control mice had negligible lesions. All mice expressing the apo B transgene developed extensive lesions, but AI KO/Btg mice developed significantly larger lesions than Btg mice: 56, 260 +/- 4630 micron 2 for AI KO/Btg (n = 27) versus 38, 120 +/- 3350 micron 2 for Btg mice (n = 19) (P < .02). Results of this study, consistent with several human epidemiological studies, indicate that HDL deficiency in the mouse does not by itself lead to the development of atherosclerosis but does increase atherosclerosis susceptibility when accompanied by other risk factors, in this case elevated LDL. PMID- 9327770 TI - The II genotype of the angiotensin-converting enzyme gene delays the onset of acute coronary syndromes. AB - The DD genotype of the angiotensin-converting enzyme (ACE) gene has been reported to be a risk factor for myocardial infarction. However, this association has not been confirmed in some populations. We hypothesized that the discrepancies between these studies may be due to their definition of ischemic heart diseases. According to the genotype of the ACE gene, we analyzed the profiles of 320 patients who underwent coronary angiography for possible ischemic heart diseases. Of these, 23 patients had no significantly diseased vessels and no acetylcholine induced vasospasm (normal acetylcholine responder [NAR]) (II, 7; ID, 14; DD, 2), 34 patients had no significantly diseased vessels and acetylcholine-induced vasospasm (paradoxical acetylcholine responder: [PAR]) (II, 15; ID, 18; DD, 1), 80 angina pectoris (AP) patients had significantly diseased vessels (II, 41; ID, 37; DD, 2), and 183 patients demonstrated myocardial infarction (MI) (II, 67; ID, 91; DD, 25). The frequency of the DD genotype was significantly lower in PAR and AP patients than in those with MI (P = .0344). Next we analyzed the length of time between the first anginal pain and the onset of myocardial infarction in the MI group. We obtained reliable information regarding this period in 149 of the 183 patients. This period was significantly shorter in the ID and DD groups than in the II group (P = .0022). Multiple regression analyses revealed that this period was significantly determined (P = .0003, R = .324) by the genotype of the ACE gene (II = 1, ID + DD = 2, P = .0003) and age (P = .034). The D allele of the ACE gene and lower age were associated with a shorter period. On the other hand, the genotype of the ACE gene had no significant effect on the number of significantly diseased (> 50%) lesions. The frequency of the D allele in subjects with a rapid progression of MI was significantly higher than that in subjects with a prolonged history of stable AP (P < .0001). In summary, the II genotype of the ACE gene was associated with a longer period of time between the first anginal pain and the onset of myocardial infarction than the ID and DD genotypes of the ACE gene. PMID- 9327771 TI - Fatty streak formation in fat-fed mice expressing human copper-zinc superoxide dismutase. AB - Studies in vitro have shown that copper-zinc superoxide dismutase (CuZn-SOD) inhibits a number of events putatively involved in atherogenesis, including cell mediated oxidation of LDL. To investigate whether increased activity of CuZn-SOD reduces atherogenesis in vivo, we examined diet-induced fatty streak formation in CuZn-SOD transgenic mice (n = 24) as compared with their nontransgenic littermates (n = 28). Transgenic animals were originally created by introduction of an EcoRI-BamHI human genomic DNA fragment containing the CuZn-SOD gene and its regulatory elements into B6SJL zygotes. For the current studies, the transgene was bred for 12 generations into the atherosclerosis-susceptible C57BL/6 background. Animals were fed atherogenic diets (15% fat, 1.25% cholesterol, 0.5% Na cholate) starting at 100 weeks of age and extending for 18 weeks. At the end of the diet period, aortic SOD activity was two-fold higher in transgenics than nontransgenics (mean +/- SE: 46.7 +/- 5.8 versus 20.1 +/- 2.4 units/mg of protein, P < .001). Levels of protein-bound amino acid oxidation products (meta-, ortho-, and dityrosine) were either similar or lower in aorta and heart from transgenics as compared with nontransgenics, suggesting that amplification of CuZn-SOD activity above the normal complement had modest inhibitory effects on basal oxidative stress in these tissues. CuZn-SOD overexpression did not reduce the extent of lesion development as analyzed by quantitative lipid staining of serial sections of the proximal aorta; mean lesion areas (+/- SE) were 997 +/- 478 and 943 +/- 221 mu 2 in transgenics and nontransgenics, respectively. Notably, the range of values for lesion area was 2.2-fold greater in transgenics (0-8403 versus 0-3868 mu 2 in nontransgenics). Moreover, within this group, lesion area showed a significant positive correlation with SOD activity (r = .611, P < .03). These results do not support an antiatherogenic effect of Cu-Zn SOD over expression and raise the possibility that high tissue SOD activity may potentiate atherogenesis in fat-fed atherosclerosis-susceptible mice [corrected]. PMID- 9327772 TI - Effects of estrus cycle, ovariectomy, and treatment with estrogen, tamoxifen, and progesterone on apolipoprotein(a) gene expression in transgenic mice. AB - Plasma levels of lipoprotein(a) (Lp(a)), are regulated by the synthetic rate of apolipoprotein(a) (apo(a)), a major protein component of this atherogenic lipoprotein. Exogenously administered sex steroid hormones are potent regulators of plasma Lp(a) concentrations. We utilized a recently developed apo(a) yeast artificial chromosome (YAC) transgenic mouse model to study the effects of ovariectomy, estrus cycle, and exogenous administration of ethinyl-estradiol, the partial estrogen receptor agonist, tamoxifen, and progesterone on circulating apo(a) plasma levels. Analysis of liver RNA revealed that estrogen and tamoxifen exerts their plasma apo(a) lowering effect at the level of apo(a) mRNA. This action of estrogen and tamoxifen may contribute to their antiatherosclerotic and cardiovascular protective effect. PMID- 9327773 TI - Inhibition of inducible nitric oxide synthase restores endothelium-dependent relaxations in proinflammatory mediator-induced blood vessels. AB - Endothelium-dependent relaxations mediated by nitric oxide (NO) are attenuated in arteries exposed to proinflammatory mediators. Because proinflammatory mediators stimulate the expression of the inducible NO synthase (iNOS) in vascular cells, the role of iNOS-derived NO in the impaired endothelium-dependent relaxation was examined in arterial ring preparations. Exposure of rabbit carotid arteries to interleukin-1 beta (IL-1 beta; 100 U/mL for 7 hours) and porcine coronary arteries to a combination of tumor necrosis factor-alpha (1000 U/mL), interferon gamma (500 U/mL), and lipopolysaccharide (10 micrograms/mL) for 15 hours (conditions that are associated with iNOS expression) markedly attenuated relaxations to receptor-dependent agonists, whereas those to the calcium ionophore A23187 and sodium nitroprusside were virtually unchanged. The impaired relaxation was not associated with a reduced level of the constitutive endothelial NOS (cNOS) but was accompanied by a reduced formation of biologically active NO as assessed in a bioassay system. The attenuated relaxation of carotid arteries to acetylcholine was not affected by superoxide dismutase and was neither found in arteries exposed to IL-1 beta for only 15 minutes nor in IL-1 beta-treated arteries for 7 hours followed by a 17-hour incubation period without the cytokine. Furthermore, no impaired relaxation was found in rings exposed to IL-1 beta in combination with either cycloheximide or N-alpha-tosyl-L-lysine chloromethyl ketone or pyrrolidine dithiocarbamate, treatments that prevent iNOS expression. In addition, selective inhibition of iNOS with S-methylisothiourea (10 mumol/L) completely restored acetylcholine-induced relaxations. These findings indicate that the continuous generation of NO induced by proinflammatory mediators plays a major role in the inhibition of endothelium-dependent relaxation, most likely by impairing a step in the signal transduction cascade that links activation of endothelial receptors to the calcium-calmodulin dependent activation of NOS. PMID- 9327774 TI - Regulation of lipoprotein metabolism by thiazolidinediones occurs through a distinct but complementary mechanism relative to fibrates. AB - Thiazolidinediones are antidiabetic agents, which not only improve glucose metabolism but also reduce blood triglyceride concentrations. These compounds are synthetic ligands for PPAR gamma, a transcription factor belonging to the nuclear receptor subfamily of PPARs, which are important transcriptional regulators of lipid and lipoprotein metabolism. The goal of this study was to evaluate the influence of a potent thiazolidinedione, BRL49653, on serum lipoproteins and to determine whether its lipid-lowering effects are mediated by changes in the expression of key genes implicated in lipoprotein metabolism. Treatment of normal rats for 7 days with BRL49653 decreased serum triglycerides in a dose-dependent fashion without affecting serum total and HDL cholesterol and apolipoprotein (apo) A-I and apo A-II concentrations. The decrease in triglyceride concentrations after BRL49653 was mainly due to a reduction of the amount of VLDL particles of unchanged lipid and apo composition. BRL49653 treatment did not change triglyceride production in vivo as analyzed by injection of Triton WR 1339, indicating a primary action on triglyceride catabolism. Analysis of the influence of BRL49653 on the expression of LPL and apo C-III, two key players in triglyceride catabolism, showed a dose-dependent increase in mRNA levels and activity of LPL in epididymal adipose tissue, whereas liver apo C-III mRNA levels remained constant. Furthermore, addition of BRL49653 to primary cultures of differentiated adipocytes increased LPL mRNA levels, indicating a direct action of the drug on the adipocyte. Simultaneous administration of BRL49653 and fenofibrate, a hypolipidemic drug that acts primarily on liver through activation of PPAR alpha both decreased liver apo C-III and increased adipose tissue LPL mRNA levels, resulting in a more pronounced lowering of serum triglycerides than each drug alone. In conclusion, both fibrates and thiazolidinediones exert a hypotriglyceridemic effect. While fibrates act primarily on the liver by decreasing apo C-III production, BRL49653 acts primarily on adipose tissue by increasing lipolysis through the induction of LPL expression. Drugs combining both PPAR alpha and gamma activation potential should therefore display a more efficient hypotriglyceridemic activity than either compound alone and may provide a rationale for improved therapy for elevated triglycerides. PMID- 9327776 TI - Behaviorally elicited heart rate reactivity and atherosclerosis in ovariectomized cynomolgus monkeys (Macaca fascicularis). AB - It has been hypothesized that atherogenesis is accelerated among individuals who exhibit heightened cardiovascular reactions to psychologic stress. We have reported previously that the coronary atherosclerosis of cholesterol-fed, male and reproductively intact (premenopausal) female cynomolgus monkeys was exacerbated in animals that experienced the largest heart rate (HR) reactions to a fear-eliciting laboratory stressor. In this article, we report a similar relationship among 20 female monkeys that were rendered estrogen-deficient (by ovariectomy) and subsequently treated with replacement of both estrogen and progesterone. At the beginning of a 30-month study period, animals were fitted with ECG telemetry devices, and their HRs were recorded under baseline and stressed conditions. Stress HR measurements were obtained during a standard challenge involving threatened capture and physical handling of the animals. As part of a related experiment, monkeys were then ovariectomized and, for the remainder of the study, administered 17 beta-estradiol (continuously) and progesterone (cyclically) by subcutaneous Silastic implant (Dow Corning). Animals consumed a cholesterol-containing diet throughout, and HR measurements were repeated in the 24th month. At necropsy, the magnitude of animals' HR responses to stress correlated significantly with intimal area measurements in the left anterior descending and circumflex coronary arteries (r = .59 and r = .57, respectively; P < .009). This association was due to a marked exacerbation of coronary atherosclerosis in animals comprising the upper third of the reactivity distribution. Although total and HDL cholesterol concentrations also covaried with HR reactivity, the greater atherosclerosis of "high" HR reactors persisted after statistical adjustment for concomitant variability in plasma lipids. HR reactivity was unrelated to blood pressure, body weight, or social behavior. PMID- 9327775 TI - Dietary fat clearance in normal subjects is modulated by genetic variation at the apolipoprotein B gene locus. AB - Apolipoprotein B (apo B) plays a dominant role in cholesterol homeostasis. Several polymorphic sites within or adjacent to the gene locus for apo B have been detected. The X+ allele (XbaI restriction site present) of the XbaI restriction fragment polymorphism on the apo B gene has been found in some studies to be associated with higher serum cholesterol and/or triglyceride levels and with greater dietary response. The present study was designed to evaluate whether the apo B XbaI polymorphism was associated with the interindividual variability observed during postprandial lipemia. Fifty-one healthy young male volunteers [20 X-/X- (X-), and 31 X+/X- or X+/X+ (X+)], homozygotes for the apo E3 allele, were subjected to a vitamin A-fat load test. Subjects with the X- genotype had significantly greater retinyl palmitate (RP) and apo B-48 postprandial responses on both the large and the small TRL lipoprotein fractions compared with X+ subjects. In summary, subjects with the X-/X- genotype at the apo B locus have a greater postprandial response than X+ subjects. These differences observed in postprandial lipoprotein metabolism could explain some of the reported associations of this polymorphism to coronary heart disease risk. PMID- 9327777 TI - Cytomegalovirus infection, lipoprotein(a), and hypercoagulability: an atherogenic link? AB - A link between cytomegalovirus (CMV) infection and atherosclerosis has been suggested by experimental, clinical, and epidemiologic studies. We investigated the association between CMV antibody titers in serum collected in 1974 in 300 adult residents in Washington County, Md, and hemostatic parameters in plasma collected in 1987 through 1989, when these individuals participated in the baseline examination of the Atherosclerosis Risk in Communities Study. The cross sectional association of CMV serum antibodies and hemostatic parameters was also explored in another set of Atherosclerosis Risk in Communities cases and controls. In the longitudinal analyses, CMV titers in 1974 were directly associated with 1987 through 1989 plasma levels of von Willebrand factor, factor VIII, and protein C and negatively associated with activated partial thromboplastin time. In the cross-sectional analyses, CMV titers were directly related to antithrombin III and fibrinogen levels. When the association between CMV antibodies and atherosclerosis was examined in stratified analyses, a significant association was restricted to individuals with high levels of lipoprotein(a) and fibrinogen. These results are compatible with previous evidence suggesting that CMV virus might have procoagulant properties. The possible synergism of CMV infection and resulting hypercoagulability with reduced fibrinolysis due to increased lipoprotein(a) levels deserves further investigation. PMID- 9327778 TI - Augmented adenovirus-mediated gene transfer to atherosclerotic vessels. AB - Vascular endothelium is an important target for gene transfer in atherosclerosis. In this study, we examined gene transfer to normal and atherosclerotic blood vessels from two species, using an organ culture method. Using normal aorta, we determined optimal dose, duration of exposure to adenovirus, and duration of incubation of vessels in tissue culture. Aortas from normal and atherosclerotic monkeys were cut into rings and incubated for 2 hours with a recombinant adenovirus, carrying the reporter gene for beta-galactosidase driven by a cytomegalovirus (CMV) promoter. After 20 hours of incubation, transgene expression was assessed with a morphometric method after histochemical staining and a chemiluminescent assay of enzyme activity. Expression of beta-galactosidase after histochemical staining, expressed as percentage of total cells, was similar in adventitial cells of normal monkeys (21 +/- 4%, mean +/- SE%) and atherosclerotic monkeys (25 +/- 12%). Transgene expression in endothelium was higher in atherosclerotic than in normal vessel (53 +/- 3% versus 27 +/- 7%, P < .05). Chemiluminescent assay indicated greater beta-galactosidase activity (2.5 +/- 0.6 mU/mg of protein) in the intima and media of atherosclerotic than normal vessels (0.6 +/- 0.2 mU/mg of protein, P < .05). Aortas from normal (n = 6) and atherosclerotic (n = 5) rabbits also were examined. Transgene expression (after histochemical staining) in endothelium was much greater in atherosclerotic than normal rabbits (39 +/- 3% versus 9 +/- 2%, P < .05) and expression in adventitial cells was similar (normal 23 +/- 2%, atherosclerotic 24 +/- 4%). Chemiluminescent assay indicated greater beta-galactosidase activity (1.2 +/- 0.4 mU/mg of protein) in the intima and media of atherosclerotic than normal vessels (0.2 +/- 0.1 mU/mg protein, P < .05). These findings suggest that an adenoviral vector with a CMV promoter provides similar transgene expression in adventitia of both normal and atherosclerotic vessels. Gene transfer to the endothelium was much more effective in atherosclerotic than in normal vessels. Thus it may be possible to achieve greater transgene expression in atherosclerotic than in normal arteries. PMID- 9327779 TI - Efficacy and safety of a new hydroxymethylglutaryl-coenzyme A reductase inhibitor, atorvastatin, in patients with combined hyperlipidemia: comparison with fenofibrate. AB - This 24-week, randomized, open-label multicenter study evaluated the efficacy and safety of atorvastatin compared with fenofibrate in the treatment of patients with combined hyperlipidemia (CHL). Following a 6-week baseline period, 84 patients with CHL were randomly assigned to either atorvastatin treatment, 10 mg QD for 12 weeks increasing to 20 mg QD for 12 weeks, or fenofibrate treatment, 100 mg TID for 24 weeks. Changes from baseline in lipid parameters were evaluated at weeks 12 and 24. At both 10- and 20-mg doses, atorvastatin treatment resulted in significantly greater reductions in LDL cholesterol, apolipoprotein (apo) B, total cholesterol, LDL-apoB, and lipoprotein-B compared to 300-mg fenofibrate treatment (P < .05). While atorvastatin also resulted in clinically significant reductions in triglyceride, VLDL cholesterol, apoB in VLDL, triglyceride in VLDL, and apoC-III and significant increases in HDL cholesterol and apoA-I levels, fenofibrate was more effective than atorvastatin in altering all these parameters. However, by significantly affecting both the cholesterol-rich and triglyceride-rich particles, atorvastatin holds promise as a lipid-regulator able to adequately treat a broad range of patients that includes those with CHL. PMID- 9327780 TI - Importance of estrogen receptors in hepatic LDL receptor regulation. AB - Treatment with pharmacological doses of estrogen is the most potent way to stimulate hepatic LDL receptor expression in vivo. The mechanism for this effect is unclear, in part because of difficulties in inducing this stimulation in vitro. A fundamental question, whether estrogen receptors (ERs) mediate this stimulation, has not been addressed. The aim of the current study was to determine the involvement of ERs in the estrogen-induced stimulation of LDL receptors. Treatment of rats with high doses of ethynylestradiol for 7 days increased the hepatic LDL receptor protein and mRNA levels four- and threefold, respectively. LDL receptor stimulation in estrogen-treated rats was not due to their reduced food intake because hepatic LDL receptor expression did not increase in rats fasted for 72 hours. Treatment with antiestrogen (tamoxifen or clomiphene) abolished the LDL receptor stimulatory effect of ethynylestradiol at both the protein and mRNA levels. Antiestrogen alone had no effect on hepatic LDL receptor expression and did not influence the strong resistance to dietary cholesterol normally present in rats. It is concluded that ERs are critically involved in the induction of hepatic LDL receptor expression by ethynylestradiol. The known role of growth hormone for the expression of hepatic ERs may therefore play a role in the modulation of the effects of estrogen on cholesterol metabolism and hepatic LDL receptors in the rat. PMID- 9327781 TI - The antiadhesive and antithrombotic effects of the nitric oxide donor SIN-1 are combined with a decreased vasoconstriction in a porcine model of balloon angioplasty. AB - Nitric oxide has been reported to modulate platelet and neutrophil interactions with the arterial wall. In this study, we investigated the effects of the nitric oxide donor 3-morpholino-sydnonimine (SIN-1) on platelet and neutrophil adhesion, and the vasomotor response, in a porcine model of angioplasty. Carotid arterial injury was produced by balloon dilation in control (n = 10) and treated (SIN-1; 10 micrograms/kg + 1 microgram/kg/min, i.v.) (n = 8) pigs. At the site of deep arterial injury, the average platelet adhesion was 53.6 +/- 11.3 x 10(6)/cm2 in the control animals and was significantly inhibited by more than 70%, to 15.1 +/- 4.1 x 10(6)/cm2 (P < .01), by SIN-1. Neutrophil adhesion was also decreased by SIN-1, from 255.9 +/- 29.7 to 101.8 +/- 19.7 x 10(3)cm2 (P < .001). Mural thrombosis was found in 12 (71%) of the 17 injured arteries in the control group but in only 2 (17%) of the 12 injured arteries in the SIN-1-treated group (P < .05). Concomitantly, SIN-1 reduced platelet and neutrophil adhesion to the site of endothelial injury distally. The internal diameter of the carotid arteries was similar between the two groups before dilation but was 40% greater at the site of endothelial injury distally in SIN-1-treated animals after dilation (P < .05), as compared with controls. Accordingly, postangioplasty vasoconstriction was significantly attenuated from 46.3 +/- 2.9% in control pigs to 32.5 +/- 4.8% (P < .05) in SIN-1-treated animals. The beneficial effects of SIN-1 were associated with inhibition of neutrophil-mediated whole blood aggregation and of neutrophil endothelium interactions. The potent antiadhesive and antithrombotic properties of SIN-1 in vivo were confirmed in ex vivo superfusion experiments. These results indicate that administration of a nitric oxide donor may be effective in preventing the acute pathophysiological responses to arterial injury by angioplasty. PMID- 9327782 TI - Decreased reverse cholesterol transport from Tangier disease fibroblasts. Acceptor specificity and effect of brefeldin on lipid efflux. AB - Tangier disease is characterized by HDL hypercatabolism and increased deposition of cholesterol in tissues. Tangier disease skin fibroblasts have decreased apoA-I mediated cholesterol and phospholipid efflux, which may lead to the excess accumulation of cellular cholesterol. The mechanism of apolipoprotein-mediated cholesterol efflux and the apolipoprotein acceptor specificity for cholesterol efflux from normal and Tangier disease fibroblasts was investigated. Normal cells readily effluxed cholesterol and phospholipid to apoA-I and to all of the other apolipoproteins tested (apoA-II, AIV, C-I, C-II, C-III). In contrast, Tangier cells were almost completely defective in cholesterol efflux to apoA-I and to all of the other apolipoproteins tested. HDL was also less effective, by approximately 50%, in stimulating cholesterol efflux from Tangier cells compared with normal cells. In addition, Tangier cells also showed significantly reduced phospholipid efflux to both apolipoproteins and HDL. A similar rate of cholesterol efflux, however, was observed from normal and Tangier cells when phospholipid vesicles or cyclodextrin were used as acceptors. In contrast to normal cells, only phospholipid vesicles and cyclodextrin and not apoA-I or HDL depleted intracellular cholesteryl esters from Tangier cells. Brefeldin, an inhibitor of intracellular vesicular trafficking, decreased HDL-mediated cholesterol efflux by approximately 40% but almost completely blocked both cholesterol and phospholipid efflux to apoA-I from normal cells. Brefeldin also inhibited cholesteryl ester depletion by apoA-I and HDL from normal cells. Brefeldin, however, had no significant effect on cholesterol efflux from Tangier cells to HDL. In summary, Tangier cells were found to be defective in both cholesterol and phospholipid efflux to HDL and apoA-I. The defect in apolipoprotein-mediated lipid efflux was not specific for apoA-I but also occurred for other apolipoproteins, and brefeldin blocked HDL-mediated lipid efflux from normal but not Tangier disease cells. On the basis of these results, a model is proposed whereby decreased cholesterol efflux by apolipoproteins in Tangier cells is the result of a defect in a brefeldin-sensitive pathway of lipid efflux. PMID- 9327783 TI - Hormonal regulation of lipoprotein(a) levels: effects of estrogen replacement therapy on lipoprotein(a) and acute phase reactants in postmenopausal women. AB - Estrogen lowers lipoprotein(a) [Lp(a)] levels, but the mechanisms involved have not been clarified. To address the relationship between estrogenic effects on Lp(a) and serum lipids, and on other plasma proteins of hepatic origin, 15 healthy postmenopausal women participated in a randomized, double-blinded, placebo-controlled, crossover study with 4 weeks of oral conjugated estrogens (0.625 mg/d) and placebo, separated by a 6-week period. Lp(a) levels decreased during estrogen treatment in 14 of the 15 subjects (mean decrease, 23%; P < .001). In response to estrogen, apolipoprotein A-I (apoA-I), HDL cholesterol, and triglyceride levels increased by 12% (P = .001), 11% (P < .001), and 10% (P = .02), respectively. Apolipoprotein B (apoB) and LDL cholesterol levels decreased by 7% (P = .01) and 12% (P = .03), respectively, ApoB, LDL cholesterol, and Lp(a) levels fell within 1 week of treatment, whereas apoA-I and HDL cholesterol levels rose more slowly. Levels of acid alpha 1-glycoprotein (AAG) and haptoglobin (HPT), two hepatically derived acute phase proteins, also decreased during estrogen treatment by 18% (P < .001) and 25% (P = .002), respectively. Although the changes in AAG and HPT in response to estrogen were highly correlated (r = .67, P = .009), we were unable to detect a correlation between change in either acute phase protein and change in Lp(a) (r = -.14 and -.24, P = .64 and .41). The lack of correlation between the changes in two acute phase reactants and Lp(a) suggests different underlying mechanisms for the effects of estrogen on these liver-derived proteins. PMID- 9327784 TI - Signal transduction through trimeric G proteins in megakaryoblastic cell lines. AB - The biogenesis of trimeric G proteins was investigated by measurement of the expression of alpha-subunits in the megakaryoblastic cell lines MEG-01, DAMI, and CHRF-288-11, representing stages of increasing maturation, and compared with platelets. Megakaryoblasts and platelets contained approximately equal amounts of Gi alpha-1/2, Gi alpha-3, Gq alpha, and G12 alpha protein. Maturation was accompanied by (1) downregulation of mRNA for Gs alpha and disappearance of iloprost-induced Ca2+ mobilization, (2) upregulation of the long form of Gs alpha protein (Gs alpha-L) and an increase in iloprost-induced cAMP formation, and (3) upregulation of G16 alpha mRNA and G16 alpha protein and appearance of thromboxane A2-induced signaling (Ca2+ mobilization and stimulation of prostaglandin I2-induced cAMP formation). Gz alpha protein was absent in the megakaryoblasts despite weak expression of Gz alpha mRNA in DAMI and relatively high levels of Gz alpha mRNA and Gz alpha protein in platelets. These findings reveal major changes in G protein-mediated signal transduction during megakaryocytopoiesis and indicate that G16 alpha couples the thromboxane receptor to phospholipase C beta. PMID- 9327785 TI - Expression of matrix metalloproteinases and their inhibitor TIMP-1 in the rat carotid artery after balloon injury. AB - The temporal relationship of matrix metalloproteinases (MMPs) and a specific tissue inhibitor (TIMP-1) has been examined by reverse transcription-polymerase chain reaction and substrate zymography, after balloon catheter angioplasty of the rat carotid artery. The contralateral uninjured carotid artery was used as a comparative control. Of the MMPs examined, only MMP-2 (72-kDa gelatinase) was produced constitutively by normal uninjured arteries. After injury, MMP-2 mRNA levels fell compared with the uninjured arteries; by 24 hours, levels had increased 2-fold. Zymography showed that the inactive form of MMP-2 predominated in uninjured vessels, but after injury, the level of the active form was increased. MMP-9 (92-kDa gelatinase) mRNA levels and activity peaked at 6 hours after injury and were still detectable at 7 days. MMP-3 (stromelysin) expression was detectable at low levels as early as 2 hours after injury and showed an approximate 2-fold increase of expression at 7 days. The presence of the active protein paralleled the mRNA expression. The inhibitor TIMP-1 mRNA was first detected 6 hours after injury and showed a marked peak of expression at 24 hours; however, no expression was detected by 7 days. The presence of a constitutively expressed, low molecular weight caseinolytic enzyme (27 kDa) was observed, and the induction of a caseinolytic enzyme (30 kDa) was noted that was induced as early as 2 hours after injury, peaked at 6 hours, and was barely detectable by 7 days. These results demonstrate that the process of extracellular matrix breakdown by MMPs after balloon catheter-induced injury is controlled by a tightly regulated temporal response by the genes responsible for the production of these enzymes and their inhibitor and by post-translational activation of the proenzymes. PMID- 9327786 TI - Putting a price on drugs. PMID- 9327787 TI - Foreign specialists need not apply. PMID- 9327788 TI - Patient confidentiality and the law. PMID- 9327789 TI - This experiment has failed. PMID- 9327790 TI - Physician payment: incentives change with supply. PMID- 9327791 TI - Hep to hepatitis C. PMID- 9327792 TI - A look at 350 years of physicians' fees in Quebec. PMID- 9327793 TI - The population explosion revisited. PMID- 9327795 TI - Costs and benefits of routine follow-up after curative treatment for endometrial cancer. AB - OBJECTIVE: To examine the costs of routine outpatient follow-up after curative treatment of endometrial cancer, and to determine whether this leads to early detection of recurrence or survival. The impact of specific disease characteristics on survival is examined. DESIGN: Retrospective chart review, and calculation of costs. SETTING: Ottawa Regional Cancer Centre-Civic Division (ORCC C). PATIENTS: All 432 patients referred to the ORCC-C with endometrial cancer between 1982 and 1991 who received treatment with curative intent and who continued with routine follow-up. RESULTS: Cancer recurred in 50 patients (11.57%). There was no statistically significant difference in overall survival between patients with symptomatic and asymptomatic recurrences, or between those with recurrences detected during routine follow-up visits or in the interval between routine visits. Of 4830 Papanicolaou (Pap) smears performed routinely, cancer was detected in 6 cases. The mean cost of the routine follow-up procedures for each patient with a recurrence was $19,200. CONCLUSION: Intensive follow-up of women with endometrial cancer does not result in improved survival. A prospective randomized study is warranted to evaluate other potential benefits of follow-up, such as improved quality of life or decreased morbidity. There is no economic or clinical justification for the routine use of the Pap smear in the follow-up of patients with endometrial cancer. The potential benefits of routine follow-up in endometrial cancer and other types of cancer with favourable prognoses warrant critical evaluation. PMID- 9327796 TI - Inappropriate hospital use by patients receiving care for medical conditions: targeting utilization review. AB - OBJECTIVE: To describe characteristics associated with inappropriate hospital use by patients in Manitoba in order to help target concurrent utilization review. Utilization review was developed to reduce inappropriate hospital use but can be a very resource-intensive process. DESIGN: Retrospective chart review of a sample of adult patients who received care for medical conditions in a sample of Manitoba hospitals during the fiscal year 1993-94; assessment of patients at admission and for each day of stay with the use of a standardized set of objective, nondiagnosis-based criteria (InterQual). PATIENTS: A total of 3904 patients receiving care at 26 hospitals. OUTCOME MEASURES: Acute (appropriate) and nonacute (inappropriate) admissions and days of stay for adult patients receiving care for medical conditions. RESULTS: After 1 week, 53.2% of patients assessed as needing acute care at admission no longer required acute care. Patients 75 years of age or older consumed more than 50% of the days of stay, and 74.8% of these days of stay were inappropriate. Four diagnostic categories accounted for almost 60% of admissions and days, and more than 50% of those days of stay were inappropriate. Patients admitted through the emergency department were more likely to require acute care (60.9%) than others (41.7%). Patients who were Treaty Indians had a higher proportion of days of stay requiring acute care than others (45.9% v. 32.8%). Patients' income and day of the week on admission (weekday v. weekend) were not predictive factors of inappropriate use. CONCLUSION: Rather than conducting a utilization review for every patient, hospitals might garner more information by targeting patients receiving care for medical conditions with stays longer than 1 week, patients with nervous system, circulatory, respiratory or digestive diagnoses, elderly patients and patients not admitted through the emergency department. PMID- 9327797 TI - Is routine follow-up after endometrial cancer justified? PMID- 9327798 TI - Measuring the appropriateness of hospital use. PMID- 9327799 TI - Adrenal incidentalomas: incidental in detection, not significance. PMID- 9327801 TI - Pheochromocytoma manifesting with shock presents a clinical paradox: a case report. AB - A 46-year-old man presented with shock and adult respiratory distress syndrome. Investigations revealed an adrenal mass that was diagnosed, by fine-needle aspiration biopsy, as pheochromocytoma. Because biopsy is contraindicated in patients with pheochromocytoma, this confusing presentation underscores the value of excluding this diagnosis by biochemical means before performing fine-needle aspiration of adrenal tumours. PMID- 9327800 TI - Report of the Canadian Hypertension Society Consensus Conference: 2. Nonpharmacologic management and prevention of hypertensive disorders in pregnancy. AB - OBJECTIVE: To provide Canadian physicians with comprehensive, evidence-based guidelines for the nonpharmacologic management and prevention of gestational hypertension and pre-existing hypertension during pregnancy. OPTIONS: Lifestyle modifications, dietary or nutrient interventions, plasma volume expansion and use of prostaglandin precursors or inhibitors. OUTCOMES: In gestational hypertension, prevention of complications and death related to either its occurrence (primary or secondary prevention) or its severity (tertiary prevention). In pre-existing hypertension, prevention of superimposed gestational hypertension and intrauterine growth retardation. EVIDENCE: Articles retrieved from the pregnancy and childbirth module of the Cochrane Database of Systematic Reviews; pertinent articles published from 1966 to 1996, retrieved through a MEDLINE search; and review of original randomized trials from 1942 to 1996. If evidence was unavailable, consensus was reached by the members of the consensus panel set up by the Canadian Hypertension Society. VALUES: High priority was given to prevention of adverse maternal and neonatal outcomes in pregnancies with established hypertension and in those at high risk of gestational hypertension through the provision of effective nonpharmacologic management. BENEFITS, HARMS AND COSTS: Reduction in rate of long-term hospital admissions among women with gestational hypertension, with establishment of safe home-care blood pressure monitoring and appropriate rest. Targeting prophylactic interventions in selected high-risk groups may avoid ineffective use in the general population. Cost was not considered. RECOMMENDATION: Nonpharmacologic management should be considered for pregnant women with a systolic blood pressure of 140-150 mm Hg or a diastolic pressure of 90-99 mm Hg, or both, measured in a clinical setting. A short-term hospital stay may be required for diagnosis and for ruling out severe gestational hypertension (preeclampsia). In the latter case, the only effective treatment is delivery. Palliative management, dependent on blood pressure, gestational age and presence of associated maternal and fetal risk factors, includes close supervision, limitation of activities and some bed rest. A normal diet without salt restriction is advised. Promising preventive interventions that may reduce the incidence of gestational hypertension, especially with proteinuria, include calcium supplementation (2 g/d), fish oil supplementation and low-dose acetylsalicylic acid therapy, particularly in women at high risk for early-onset gestational hypertension. Pre-existing hypertension should be managed the same way as before pregnancy. However, additional concerns are the effects on fetal well-being and the worsening of hypertension during the second half of pregnancy. There is, as yet, no treatment that will prevent exacerbation of the condition. VALIDATION: The guidelines share the principles in consensus reports from the US and Australia on the nonpharmacologic management of hypertension in pregnancy. PMID- 9327803 TI - Cochlear implants: the head-on collision between medical technology and the right to be deaf. AB - The debate over using cochlear implants to help deaf people communicate with those who can hear continues to rage. Some have welcomed the new technology, but others say the deaf have their own culture and do not need to be "cured". PMID- 9327802 TI - Preventing influenza outbreaks in long-term care facilities. PMID- 9327804 TI - Deportation proceedings against Canadian MDs may hold lesson for others heading south. AB - Two Alberta physicians who emigrated to a medically underserviced part of Kentucky have learned a harsh lesson about American immigration law. Drs. David Zetter and Sabina Seitz had been settled in western Kentucky for 2 years when the US government launched deportation proceedings against them. American officials allege that they misrepresented themselves when they entered the US on a temporary visa. They may be allowed to stay following a public outcry against their deportation. PMID- 9327805 TI - "Take some action, take some risk," conference on rural recruiting told. AB - Physicians attending a recent conference on the retention of physicians in rural areas proposed more than 40 recommendations for dealing with recruitment and retention issues. The conference was organized by Ontario's residents, who have been feeling the impact of attempts to encourage physicians to move to rural and underserviced areas. PMID- 9327806 TI - Looking back and looking ahead as the CMA turns 130. AB - The handful of "medical men" who founded the CMA 130 years ago this month could not have predicted the changes their association would witness. In this article Dr. John Bennett discusses some of these changes and considers some of the issues that may lie ahead. PMID- 9327807 TI - Cost of malpractice protection on rise in UK, too. AB - As in Canada, medical malpractice premiums in the United Kingdom are on the rise. In recent years there has been a 15%-20% annual rise in the cost of claims, and litigation costs for the National Health Service are soaring. Now, reports Caroline Richmond, another surge of litigation may be on the horizon because a 1996 change makes it possible for lawyers to take cases on a contingency basis. PMID- 9327808 TI - The effect of home care on hospital days: what is the take home message? PMID- 9327809 TI - A memorial. Professor Sylvester E. Berki, 1930-1996. PMID- 9327810 TI - The effects of predetermined payment rates for Medicare home healthcare. AB - OBJECTIVE: To assess the effects of an alternative method of paying home health agencies for services to Medicare beneficiaries, based on a demonstration program. DATA SOURCES/STUDY SETTING: Primary and secondary data collected on participating home health agencies in five states and their patients during the three-year demonstration period. Primary data included patient surveys at discharge and six months later, and two rounds of interviews with executive staff of the agencies. Secondary data included agencies' Medicare cost reports, quality assurance reviews, Medicare claims data, demonstration claims data, demonstration patient intake forms, and plan of treatment forms. STUDY DESIGN: The 47 agencies volunteering to participate in the demonstration were each randomly assigned to the treatment or control group. Treatment group agencies were paid a predetermined rate based on their inflation-adjusted cost per visit during the year preceding the demonstration; control group agencies were paid under Medicare's conventional cost reimbursement method. Demonstration impacts were estimated by comparing outcomes for the two groups of agencies and their respective patients, using regression models to control for any remaining differences. PRINCIPAL FINDINGS: Agencies paid under prospective rate setting were slightly better at holding per-visit cost increases below inflation than were control group agencies. The change in payment method had no effect on agencies' volume of Medicare visits or quality of care, nor on patients' use of Medicare services or other formal or informal care services. CONCLUSION: Changing from cost-based reimbursement to predetermined payment rates for Medicare home healthcare visits would not lead to large savings for the Medicare program, but would not increase costs to Medicare or adversely affect patients or their caregivers. PMID- 9327812 TI - Differences across payors in charges for agency-based home health services: evidence from the National Home and Hospice Care Survey. AB - OBJECTIVE: To investigate charge and payment differentials for home health services across different payors. DATA SOURCES: The 1992 National Home and Hospice Care Survey, a nationally representative survey of home and hospice care agencies and their patients, collected by the National Center for Health Statistics. STUDY DESIGN: We compare the average charge for a Medicare home health visit to the average charge for patients with other sources of payment. In making such comparisons, we control for differences across payors in service mix and agency characteristics. PRINCIPAL FINDINGS: Agencies charge various payors different amounts for similar services, and Medicare is consistently charged more than other payors. CONCLUSIONS: Findings imply the potential existence of payment differentials across payors for home health services, with Medicare and privately insured patients likely to be paying more than others for similar services. Such conclusions raise the possibility that, as in other segments of the healthcare market, cost-shifting and price discrimination might exist within the home health industry. Future research should explore these issues, along with the question of whether Medicare is paying too much for home health services. PMID- 9327811 TI - Impact of home care on hospital days: a meta analysis. AB - OBJECTIVE: To examine the impact of home care on hospital days. DATA SOURCES: Search of automated databases covering 1964-1994 using the key words "home care," "hospice," and "healthcare for the elderly." Home care literature review references also were inspected for additional citations. STUDY SELECTION: Of 412 articles that examined impact on hospital use/cost, those dealing with generic home care that reported hospital admissions/cost and used a comparison group receiving customary care were selected (N = 20). STUDY DESIGN: A meta-analytic analysis used secondary data sources between 1967 and 1992. DATA EXTRACTION: Study characteristics that could have an impact on effect size (i.e., country of origin, study design, disease characteristics of study sample, and length of follow-up) were abstracted and coded to serve as independent variables. Available statistics on hospital days necessary to calculate an effect size were extracted. If necessary information was missing, the authors of the articles were contacted. METHODS: Effect sizes and homogeneity of variance measures were calculated using Dstat software, weighted for sample size. Overall effect sizes were compared by the study characteristics described above. PRINCIPAL FINDINGS: Effect sizes indicate a small to moderate positive impact of home care in reducing hospital days, ranging from 2.5 to 6 days (effect sizes of -.159 and -.379, respectively), depending on the inclusion of a large quasi-experimental study with a large treatment effect. When this outlier was removed from analysis, the effect size for studies that targeted terminally ill patients exclusively was homogeneous across study subcategories; however, the effect size of studies that targeted nonterminal patients was heterogeneous, indicating that unmeasured variables or interactions account for variability. CONCLUSION: Although effect sizes were small to moderate, the consistent pattern of reduced hospital days across a majority of studies suggests for the first time that home care has a significant impact on this costly outcome. PMID- 9327813 TI - The demand for health insurance coverage by low-income workers: can reduced premiums achieve full coverage? AB - OBJECTIVE: To assess the degree to which premium reductions will increase the participation in employer-sponsored health plans by low-income workers who are employed in small businesses. DATA SOURCES/STUDY SETTING: Sample of workers in small business (25 or fewer employees) in seven metropolitan areas. The data were gathered as part of the Small Business Benefits Survey, a telephone survey of small business conducted between October 1992 and February 1993. STUDY DESIGN: Probit regressions were used to estimate the demand for health insurance coverage by low-income workers. Predictions based on these findings were made to assess the extent to which premium reductions might increase coverage rates. DATA COLLECTION/EXTRACTION METHODS: Workers included in the sample were selected, at random, from a randomly generated set of firms drawn from Dun and Bradstreet's DMI (Dun's Market Inclusion). The response rate was 81 percent. FINDINGS: Participation in employer-sponsored plans is high when coverage is offered. However, even when coverage is offered to employees who have no other source of insurance, participation is not universal. Although premium reductions will increase participation in employer-sponsored plans, even large subsidies will not induce all workers to participate in employer-sponsored plans. For workers eligible to participate, subsidies as high as 75 percent of premiums are estimated to increase participation rates from 89.0 percent to 92.6 percent. For workers in firms that do not sponsor plans, similar subsidies are projected to achieve only modest increases in coverage above that which would be observed if the workers had access to plans at unsubsidized, group market rates. CONCLUSIONS: Policies that rely on voluntary purchase of coverage to reduce the number of uninsured will have only modest success. PMID- 9327815 TI - Promoting clinical involvement in hospital quality improvement efforts: the effects of top management, board, and physician leadership. AB - STUDY QUESTION: An examination of the effects of top management, board, and physician leadership for quality on the extent of clinical involvement in hospital CQI/TQM efforts. DATA SOURCES: A sample of 2,193 acute care community hospitals, created by merging data from a 1989 national survey on hospital governance and a 1993 national survey on hospital quality improvement efforts. STUDY DESIGN: Hypotheses were tested using Heckman's two-stage modeling approach. Four dimensions of clinical involvement in CQI/TQM were examined: physician participation in formal QI training, physician participation in QI teams, clinical departments with formally organized QA/QI project teams, and clinical conditions and procedures for which quality of care data are used by formally organized QA/QI project teams. Leadership measures included CEO involvement in CQI/TQM, board quality monitoring, board activity in quality improvement, active staff physician involvement in governance, and physician-at-large involvement in governance. Relevant control variables were included in the analysis. PRINCIPAL FINDINGS: Measures of top management leadership for quality and board leadership for quality showed significant, positive relationships with measures of clinical involvement in CQI/TQM. Active-staff physician involvement in governance showed positive, significant relationships with clinical involvement measures, while physician-at-large involvement in governance showed significant, negative relationships. CONCLUSIONS: Study results suggest that leadership from the top promotes clinical involvement in CQI/TQM. Further, results indicate that leadership for quality in healthcare settings may issue from several sources, including managers, boards, and physician leaders. PMID- 9327814 TI - A controlled letter intervention to change prescribing behavior: results of a dual-targeted approach. AB - OBJECTIVE: To determine the effectiveness of a drug utilization review (DUR) letter intervention sent only to physicians, sent only to pharmacists, or sent to both physicians and pharmacists in changing physician prescribing behavior for dipyridamole. DATA SOURCES/STUDY SETTING: A Wisconsin Medicaid prescription drug database for data from March 1991 through May 1992 related to both long-term care and ambulatory patient settings. STUDY DESIGN: The effects of a DUR letter intervention were tested using a field study, pre-post, nonequivalent control group, quasi-experimental design. The effects of the letter intervention in terms of dipyridamole expenditures (dollars reimbursed to pharmacies by Medicaid), expenditures for related drugs (aspirin, ticlopidine, sulfinpyrazone) and numbers of patients for whom dipyridamole was discontinued were examined across three experimental groups and a control group. DATA COLLECTION/EXTRACTION METHODS: Dipyridamole expenditures for each study patient during a six-month preintervention and six-month postintervention period were collected from Medicaid prescription drug claims. Patients who had zero dipyridamole expenditures throughout the six-month postintervention period were classified as having had dipyridamole discontinued. PRINCIPAL FINDINGS: Letters sent to both physicians and pharmacists resulted in a greater percentage of patients discontinuing dipyridamole relative to controls and statistically significant differences in postintervention dipyridamole expenditures relative to controls in both the long-term care and ambulatory patient populations. CONCLUSIONS: Interventions that focus on another person in the drug use process in addition to the physician may have greater effects on a change in the prescribing of a targeted drug than letters to physicians alone. PMID- 9327816 TI - Adjusting cesarean delivery rates for case mix. AB - OBJECTIVES: (1) To describe the issues in developing a clinical predictor of cesarean delivery that could be used to adjust reported cesarean rates for case mix, and (2) to compare its performance to other, simpler predictors using clinical and statistical criteria. DATA SOURCES: Singleton births greater than 2,500 grams in Washington State in 1989 and 1990 for whom mothers and infant hospital discharge records could be matched to birth certificate data. DESIGN: Statistical analysis of retrospective merged hospital and birth certificate data, which were used to develop variables and models to predict the probability that any particular delivery would be a cesarean. PRINCIPAL FINDINGS: Merged data led to better predictor variables than those based on one source. A simple four category hierarchical classification into births with prior cesarean, breech but no prior cesarean, first birth, and other explains 30 percent of the variance in individual cesarean rates. The full clinical model fit the data well and explained 37 percent of the variance. Multiparas without serious complications comprised 35 percent of the mothers and averaged less than 2 percent cesareans. A hospital's predicted cesarean rate depends strongly on the proportion of its births that are first births. CONCLUSION: Government and private agencies have reported cesarean rates as measures of hospital performance. Depending on data and resources available, both simple and complex measures of case mix can be used to adjust reported rates. These adjustments should not include all variables related to the rates. Proper adjustments may not alter hospital rankings greatly, but they will improve the validity and acceptability of the reports. PMID- 9327817 TI - Development of function-related groups version 2.0: a classification system for medical rehabilitation. AB - OBJECTIVE: To present a new version (2.0) of the Functional Independence Measure Function Related Group (FIM-FRG) case-mix measure. DATA SOURCE/STUDY SETTING: 85,447 patient discharges from 252 freestanding facilities and hospital units contained in the 1992 Uniform Data System for Medical Rehabilitation. STUDY DESIGN: Patient impairment category, functional status at admission to rehabilitation, and patient age were used to develop groups that were homogeneous with respect to length of stay. Within each impairment category patients were randomly assigned to one data set to create the system (through recursive partitioning) or a second set for validation. Clinical and statistical criteria were used to increase the percentage of patients classified, expand the impairment categories of FIM-FRGs Version 1.1, and evaluate the incremental predictive ability of coexisting medical diagnoses. Predictive stability over time was evaluated using 1990 discharges. PRINCIPAL FINDINGS: In Version 2.0, the percentage of patients classified was increased to 92 percent. Version 2.0 includes two new impairment categories and separate groups for patients admitted to rehabilitation for evaluation only. Coexisting medical diagnoses did not improve LOS prediction. The system explains 31.7 percent of the variance in the logarithm of LOS in the 1992 validation sample, and 31.0 percent in 1990 discharges. CONCLUSIONS: FIM-FRGs Version 2.0 includes more specific impairment categories, classifies a higher percentage of patient discharges, and appears sufficiently stable over time to form the basis of a payment system for inpatient medical rehabilitation. PMID- 9327818 TI - The effects of a home exercise program on impairment and health-related quality of life in persons with chronic peripheral neuropathies. AB - BACKGROUND AND PURPOSE: The effects of a home exercise program for persons with chronic peripheral neuropathies (CPN) have not been documented. We compared changes in impairment and health-related quality of life (HRQL) between exercise and control groups, investigated the relationship between HRQL and measures of impairment, and contrasted the HRQL of individuals with CPN to that previously described for the general population. SUBJECTS: Twenty-eight subjects with CPN, aged 23 to 84 years (mean = 56.2, SD = 14.9), completed the study. METHODS: Impairment measures included average muscle score (AMS), handgrip force, walking time, and forced vital capacity. The HRQL instrument measured the eight scales of the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the component scales. The exercise group (n = 14) completed a 6-week home exercise program. The control group (n = 14) did not participate in a home exercise program. RESULTS: There was an increase in the AMS in the exercise group compared with the control group. No other between-group differences were found. The exercise group improved in scores on the role limitation scales of the SF-36. The AMS and walking time were moderately correlated with the physical function scale of the SF-36 (r = .55 and -.62, respectively). The SF-36 scores of individuals with CPN were lower than scores previously described for the general population. CONCLUSION AND DISCUSSION: The home exercise program appears to be an important component of the treatment of persons with CPN. Compared with the general population, patients with CPN appear to have a lower HRQL, but some areas appear to improve following a home exercise program. PMID- 9327819 TI - Physical therapy utilization by patients with acute low back pain. AB - BACKGROUND AND PURPOSE: The purposes of this study were (1) to describe the demographic and clinical characteristics of a group of patients with acute low back pain (LBP), (2) to describe those patients who were being treated by physical therapists, and (3) to analyze their use of physical therapy services. SUBJECTS: The study sample consisted of 1,580 patients with acute LBP who were treated by 208 practitioners in North Carolina. The initial providers were primary care physicians, chiropractors, orthopedic surgeons, and primary care providers at a health maintenance organization. METHODS: A telephone interview was conducted after the initial office visit to assess demographics and medical history, health care services utilization, and functional status. Follow-up telephone interviews were also conducted 2, 4, 8, 12, and 24 weeks later. RESULTS: One hundred ninety-nine (12.6%) of the subjects reported that they saw a physical therapist either by any provider referral or by direct access. Therapeutic exercise was the most commonly reported treatment procedure. Post high-school education, receipt of Workers' Compensation, prior physical therapy for LBP, LBP and pain below the knee in one or both legs, and a higher baseline Roland-Morris Questionnaire score were associated with being treated by physical therapists. CONCLUSION AND DISCUSSION: In this study, physical therapists were utilized in the treatment of patients with greater severity of LBP. The findings demonstrate the importance of controlling for baseline characteristics when comparing outcomes of LBP when treated by different types of providers. PMID- 9327820 TI - Comparison of upper-extremity balance tasks and force platform testing in persons with hemiparesis. AB - BACKGROUND AND PURPOSE: The purpose of this study was to investigate the relationship between clinically accessible functional balance tools and sophisticated force platform measures in a standing position. SUBJECTS: Twenty persons who had hemiparesis secondary to a stroke and were ambulatory (mean age = 57.9 years, SD = 13.6, range = 35-79) were evaluated during a single testing session. METHODS: Performances on self-generated upper-extremity balance tasks using the nonparetic side (Functional Reach Test [FRT], arm raise and arm reach tasks) were compared with responses to external perturbations on the Balance System (postural sway, symmetry of weight distribution). RESULTS: No relationship was found between the upper-extremity balance tests and the force platform measures of postural sway. After suppressing the effect of age by means of partial correlation coefficients, the FRT was correlated with measures of postural symmetry in parallel stance on the Balance System (r = .66-.78). The FRT was only moderately correlated with the arm raise and arm reach tasks (r = .43 and .44). CONCLUSION AND DISCUSSION: Postural sway in response to force platform perturbations may have little relation to the postural control necessary for self generated upper-extremity balance tasks. In contrast, the FRT and the force platform measures of postural symmetry appear to be evaluating comparable standing-balance abilities in persons with hemiparesis. The modest relationship between the FRT and the arm raise and arm reach tasks limits the finding's clinical relevance. PMID- 9327821 TI - Are patellofemoral pain and quadriceps femoris muscle torque associated with locomotor function? AB - BACKGROUND AND PURPOSE: The purpose of this investigation was to determine the influence of pain and muscle weakness on gait variables in subjects with patellofemoral pain (PFP). SUBJECTS: Nineteen female subjects with a diagnosis of PFP and 19 female subjects without PFP participated in the study. METHODS: Subjects underwent gait analysis (stride characteristics and joint motion) during level walking, ascending and descending stairs, and ascending and descending ramps, in addition to isometric torque testing of the knee extensors of the involved limb. Pain and functional status also were assessed. RESULTS: Compared with the comparison group, the primary gait compensation in the PFP group was a reduced walking speed, which was a function of both a reduced stride length and cadence. Knee extensor torque was the only predictor of gait function, with increased torque correlating with improved stride characteristics. In addition, PFP was not associated with locomotor function. CONCLUSION AND DISCUSSION: These findings suggest that functional ability in persons with PFP is associated with increased quadriceps femoris muscle torque. Future research is needed to determine whether function improves with quadriceps femoris muscle strengthening. PMID- 9327822 TI - The shoulder pain and disability index: the construct validity and responsiveness of a region-specific disability measure. AB - BACKGROUND AND PURPOSE: The purposes of this study were (1) to assess the construct validity of the Shoulder Pain and Disability Index (SPADI) and (2) to determine whether the SPADI is more responsive than the Sickness Impact Profile (SIP), a generic health status measure. SUBJECTS: The sample consisted of 94 patients who were diagnosed with a shoulder problem and referred to six outpatient physical therapy clinics. METHODS: Clinically meaningful change was determined by use of an ordinal rating scale designed to determine whether the patient's shoulder function was improved, the same, or worse following treatment. Spearman rho correlations were calculated for the initial visit SPADI and SIP scores. The standardized response mean (SRM) was used to measure responsiveness for the patients who were judged to be improved. One-tailed paired t tests (alpha = .01) were used to determine whether differences existed among SRM values. RESULTS: Correlations between the SPADI and SIP scores ranged from r = .01 to r = .57. The SRM value was higher for the SPADI total score (SRM = 1.38) than for the SIP total score (SRM = 0.79). CONCLUSION AND DISCUSSION: Most correlations between SPADI and SIP scores provided support for the construct validity of the SPADI. The SPADI does not appear to strongly reflect occupational and recreational disability and is more responsive than the SIP. PMID- 9327823 TI - The effect of time and frequency of static stretching on flexibility of the hamstring muscles. AB - BACKGROUND AND PURPOSE: Frequency and duration of static stretching have not been extensively examined. Additionally, the effect of multiple stretches per day has not been evaluated. The purpose of this study was to determine the optimal time and frequency of static stretching to increase flexibility of the hamstring muscles, as measured by knee extension range of motion (ROM). SUBJECTS: Ninety three subjects (61 men, 32 women) ranging in age from 21 to 39 years and who had limited hamstring muscle flexibility were randomly assigned to one of five groups. The four stretching groups stretched 5 days per week for 6 weeks. The fifth group, which served as a control, did not stretch. METHODS: Data were analyzed with a 5 x 2 (group x test) two-way analysis of variance for repeated measures on one variable (test). RESULTS: The change in flexibility appeared to be dependent on the duration and frequency of stretching. Further statistical analysis of the data indicated that the groups that stretched had more ROM than did the control group, but no differences were found among the stretching groups. CONCLUSION AND DISCUSSION: The results of this study suggest that a 30-second duration is an effective amount of time to sustain a hamstring muscle stretch in order to increase ROM. No increase in flexibility occurred when the duration of stretching was increased from 30 to 60 seconds or when the frequency of stretching was increased from one to three times per day. PMID- 9327824 TI - Intraobserver and interobserver reliability of assessments of impairments and disabilities. AB - BACKGROUND AND PURPOSE: The purpose of this study was to evaluate the interobserver and intraobserver reliability of assessments of impairments and disabilities. SUBJECTS AND METHODS: One physical therapist's assessments were examined for intraobserver reliability. Judgments of two pairs of therapists were used to examine interobserver reliability. Reliability was assessed by Cohen's kappa. RESULTS: Of the 42 impairments and disabilities assessed by the physical therapist in the intraobserver reliability study, kappa values could be calculated for 33 items. For 31 items (94%), kappa values ranged from .40 to .91, and 2 items (6%) had kappa values of less than .40. To determine interobserver reliability, 37 items were assessed in one practice. Kappa values could be calculated for 34 items, with 30 items (88%) having kappa values ranging from .41 to .80 and 4 items (12%) showing "poor" agreement. In the second practice, 47 items were assessed for interobserver reliability. Kappa values could be calculated for 40 items, with 11 items (27.5%) having kappa values ranging from .41 to .84. Poor agreement was shown for the remaining 29 items (72.5%). CONCLUSION AND DISCUSSION: Assessments of impairments and disabilities are potentially reliable. The differences between practices of the interobserver reliability study can be explained by the fact that one of the therapists did not receive training in the use of the assessment form. More generalizable conclusions will require further study with more subjects and therapists. PMID- 9327825 TI - Maximizing functional recovery. PMID- 9327826 TI - The molecular bases of restenosis. AB - Studies on the fundamental mechanisms of restenosis after arterial intervention have undergone upheaval. The rat carotid artery has provided perhaps the most widely used experimental preparation for the study of the response to acute arterial injury and gave rise to the multiwave model of restenosis. However, simple arterial injury differs substantially from angioplasty to diseased human vessels. The human intimal lesion already contains abundant smooth muscle cells and leukocytes. Therefore, although the rat carotid injury preparation has provided keen insight into the biology of the response to injury, it mimics human restenosis poorly, if at all. The first wave and second wave of the response to arterial injury so well delineated in rats (medial smooth muscle cell proliferation and migration from media into intima) may not apply to clinical angioplasty injury at all. Smooth muscle proliferation, a constant in injury to animal arteries, may be indolent in human restenosis. Accumulation of extracellular matrix probably explains in part the dissociation between intimal thickening and smooth muscle replication. Also, failure to "remodel" outwards to maintain the increased caliber produced by interventional arterial dilatation, may prove more important than intimal thickening in determining ultimate intimal caliber. Despite a rather disappointing decade of translation of laboratory findings to the clinic, some grounds for optimism in approaching the complications of arterial intervention have emerged. Our appreciation for the biological complexities underlying the restenosis problem have increased. In interventional cardiology we can now proceed, more experienced and wiser, to longer range solutions to help our patients based on mechanistic insights. PMID- 9327827 TI - The biology of restenosis. AB - Recent studies have allowed a better understanding of the biology of restenosis. Neointimal thickening--also referred to as neointimal hyperplasia--occurs in response to experimental arterial injury. This process involves different steps which include smooth muscle cell activation, proliferation, and migration, and the production of extracellular matrix. Neointimal thickening has been identified as one of the mechanisms of restenosis after balloon angioplasty in humans. The factors which control neointimal thickening include growth factors, hormonal factors, and mechanical factors. Delinquent reendothelialization has been shown to have a permissive impact on smooth muscle cell proliferation. In addition to neointimal thickening, arterial remodeling also plays a major role in restenosis. Studies performed on animals and in human subjects have established the potential for "constrictive remodeling" to reduce the vessel lumen after angioplasty. Restenosis thus appears as a multifactorial entity that may be addressed in the future by a combined mechanical and pharmacological approach. PMID- 9327828 TI - The role of proto-oncogenes in coronary restenosis. AB - Arterial injury results in exposure of medial smooth muscle cells and adventitial fibroblasts to multiple growth factors that bind to specific cell surface receptors. These in turn activate second messengers and induce expression of immediate-early genes within minutes to hours after ligand binding to the receptor. Activation of the immediate-early genes results in passage of the stimulated cell from its nonproliferating, quiescent G0 state to the first phase of the cell cycle (G1). Coordination of the events that occur during the cell cycle is effected by a series of cyclin-dependent kinases and requires inactivation of several "tumor suppressor genes," including p53, p21, p16, p15, p27, and the retinoblastoma gene Rb, that inhibit the kinase activity of the cyclin/Cdk complexes. An understanding of the factors that regulate signal transduction, cell cycle progression, and programmed cell death has suggested several novel therapeutic strategies including (1) antisense oligonucleotide inhibition of proto-oncogene expression, (2) the use of molecular decoys or pharmacological therapies to block specific steps required for cell cycle progression, and (3) gene transfer of tumor suppressor genes. The apparent success of several of these strategies in animal models of restenosis suggests that these molecular therapies may play a valuable role in preventing intimal hyperplasia and restenosis after balloon angioplasty and vascular stenting. PMID- 9327829 TI - Remodeling of the coronary artery after vascular injury. AB - Vascular injury, such as angioplasty, induces a complex healing process that is analogous to generalized wound healing. Following initial injury, platelet thrombi and an acute inflammatory response occur, followed by cellular inhibition by monocytes/macrophages, proliferation of vascular smooth muscle cells, and extracellular matrix deposition. The injury repair concludes with organization of the matrix and reendothelialization. Recent studies suggest that restenosis or renarrowing after vascular injury is due to two components: a cellular proliferative process leading to intimal hyperplasia and remodeling of the vessel with a change in vessel geometry. Animal and clinical studies have shown that at least 60% of restenosis can be explained by unfavorable remodeling with vessel constriction rather than intimal hyperplasia. Although a number of factors appear to be associated with remodeling, alterations in extracellular matrix metabolism may be most important. PMID- 9327830 TI - Pharmacological approaches for the prevention of restenosis after percutaneous coronary intervention. AB - A large number of drug trials for prevention of restenosis have been conducted with many showing little or conflicting benefit. Antiplatelets such as aspirin, ticlopidine and thromboxane A2 receptor inhibitors have not shown a clear benefit. Similarly, antithrombotics, either acting indirectly such as heparin, or as direct thrombin inhibitors such as hirudin and hirulog, do not prevent restenosis. Trials with ACE inhibitors, HMG-CoA reductase inhibitors and fish-oil supplements have yielded inconclusive results. The antiproliferatives, angiopeptin, trapidil and tranilast have shown some benefit in small-scale studies. Other drug classes of potential benefit include the glycoprotein IIb/IIIa receptor antagonists, inhibitors of the early coagulation cascade, calcium channel blockers and nitric oxide donors. Drug research into restenosis prevention has been hampered by problems with the definition of restenosis and the applicability in humans of animal models. Although no single drug has conclusively proven effective yet, the promise of a number of agents, together with other nonpharmacological strategies will likely result in further reductions in the incidence of restenosis. PMID- 9327831 TI - Stenting for ischemic heart disease. AB - This article examines current indications to intracoronary stenting for percutaneous cardiac revascularization. The data sources are a review of the published English literature, with focus on clinical and randomized trails of coronary artery stenting. Stents reduce the need for emergency bypass surgery in the setting of acute or threatened vessel closure after percutaneous transluminal coronary angioplasty (PTCA). In selected cases, when deployed electively in large native vessels with focal stenosis, clinical and angiographic recurrence are significantly reduced. Preliminary studies have also suggested a potential role in the treatment of saphenous vein graft lesions and restenotic lesions, and in improving on suboptimal results in lesions refractory to balloon dilatation. To prevent stent thrombosis, adjunctive antiplatelet therapy has been shown to be more efficacious than anticoagulation, and optimal stent apposition to the vessel wall appears to be critical. A recent exponential increase in the use of coronary stents has revolutionized the contemporary practice of interventional cardiology. Although technical factors and feasibility have been well refined and shown, evidence-based practice is lacking for many patient subsets. PMID- 9327832 TI - Local drug delivery: current applications. AB - Catheter-based treatment of coronary artery disease has historically been based on expansion or ablating vessels. Only in the last 3 years have we had devices that allow us to choose the location to apply agents directly onto or into the arterial wall. Previous trials of pharmaceutical agents in humans have failed despite animal trials showing efficacy. These agents were given in high systemic doses of drugs that may have toxic side effects with minimal effect at the site of arterial injury. Percutaneous interventions are still limited by our need to treat thrombus, alter or passivate the arterial wall, and deliver new treatments into or through the arterial wall. This review discusses the current designs of delivery catheters, ongoing trials of locally delivered agents and gene therapy. The limits of our current understanding of delivery location and efficiency as well as future investigations are also discussed. PMID- 9327833 TI - Catecholamine synthesis and release. Overview. PMID- 9327834 TI - The effect of phosphorylation at Ser-40 on the structure and thermal stability of tyrosine hydroxylase. PMID- 9327835 TI - Factors affecting adrenal medullary catecholamine biosynthesis and release. PMID- 9327836 TI - Regulation of tyrosine hydroxylase by neuropeptides. PMID- 9327837 TI - Regulation of tyrosine hydroxylase gene expression by transsynaptic mechanisms and cell-cell contact. PMID- 9327838 TI - A new regulatory protein of catecholamine synthesizing-enzyme expression. PMID- 9327839 TI - Unique and cell-type-specific tyrosine hydroxylase gene expression. PMID- 9327840 TI - Triple colocalization of tyrosine hydroxylase, calretinin, and calbindin D-28k in the periventricular-hypophyseal dopaminergic neuronal system. PMID- 9327841 TI - Genetic disorders involving recycling and formation of tetrahydrobiopterin. PMID- 9327842 TI - Genetic basis of dominant dystonia. PMID- 9327843 TI - Mutations in the tyrosine hydroxylase gene cause various forms of L-dopa responsive dystonia. PMID- 9327844 TI - Catecholamine biosynthetic enzyme expression in neurological and psychiatric disorders. PMID- 9327845 TI - Multiple pathways in regulation of dopamine beta-hydroxylase. PMID- 9327846 TI - Examining adrenergic roles in development, physiology, and behavior through targeted disruption of the mouse dopamine beta-hydroxylase gene. PMID- 9327847 TI - Genetic diseases of hypotension. PMID- 9327848 TI - Dopamine beta-hydroxylase deficiency associated with mutations in a copper transporter gene. PMID- 9327849 TI - Glucocorticoid-phenylethanolamine-N-methyltransferase interactions in humans. PMID- 9327850 TI - Determinants of phenylethanolamine-N-methyltransferase expression. PMID- 9327851 TI - Neural control of phenylethanolamine-N-methyltransferase via cholinergic activation of Egr-I. PMID- 9327852 TI - Synexin (annexin VII) hypothesis for Ca2+/GTP-regulated exocytosis. PMID- 9327853 TI - Monoamine transmitter release from small synaptic and large dense-core vesicles. PMID- 9327854 TI - Calcium channels for exocytosis in chromaffin cells. PMID- 9327856 TI - Neurotransmitter release at individual sympathetic varicosities, boutons. AB - The secretosome hypothesis has been studied at autonomic nerve terminals. Evidence is presented that there is a nonuniform secretion probability at different release sites of these terminals. This can be correlated with their influx of calcium ions following an impulse, as expected according to the secretosome hypothesis. PMID- 9327857 TI - Appropriate target cells are required for maturation of neurotransmitter release function of sympathetic neurons in culture. PMID- 9327855 TI - Characteristics of transmitter secretion from individual sympathetic varicosities. PMID- 9327858 TI - Effects of neuropeptide Y at sympathetic neuroeffector junctions. PMID- 9327859 TI - Strategies for receptor control of neurotransmitter release. PMID- 9327860 TI - Pattern of adenosine triphosphate and norepinephrine release and clearance: consequences for neurotransmission. PMID- 9327861 TI - Corelease of noradrenaline and adenosine triphosphate from sympathetic neurones. PMID- 9327862 TI - Neuropeptide Y: an adrenergic cotransmitter, vasoconstrictor, and a nerve-derived vascular growth factor. PMID- 9327863 TI - Neuropeptide Y: a cardiac sympathetic cotransmitter? PMID- 9327864 TI - Biochemistry of somatodendritic dopamine release in substantia nigra: an in vivo comparison with striatal dopamine release. PMID- 9327865 TI - The use of dual-probe microdialysis for the study of catecholamine release in the brain and pineal gland. PMID- 9327866 TI - Kinetics and geometry of the excitatory dopaminergic transmission in the rat striatum in vivo. PMID- 9327867 TI - In vivo and in vitro assessment of dopamine uptake and release. PMID- 9327868 TI - Catecholamine reuptake and storage. Overview. PMID- 9327869 TI - Molecular physiology and regulation of catecholamine transporters. PMID- 9327870 TI - Localization of dopamine transporter protein by microscopic histochemistry. PMID- 9327871 TI - Cellular and subcellular localization of the dopamine transporter in rat cortex. PMID- 9327873 TI - Inactivation of the dopamine transporter reveals essential roles of dopamine in the control of locomotion, psychostimulant response, and pituitary function. PMID- 9327872 TI - Cloned catecholamine transporters expressed in polarized epithelial cells: sorting, drug sensitivity, and ion-coupling stoichiometry. PMID- 9327874 TI - Role of protein kinase C and second messengers in regulation of the norepinephrine transporter. PMID- 9327875 TI - Electrophysiological analysis of transporter function. PMID- 9327876 TI - Voltammetric approaches to kinetics and mechanism of the norepinephrine transporter. PMID- 9327878 TI - Modulation of quantal dopamine release by psychostimulants. PMID- 9327877 TI - Voltage-dependency of the dopamine transporter in rat brain. PMID- 9327879 TI - Regulation of dopamine transporter mRNA levels in the central nervous system. PMID- 9327880 TI - Structural diversity in the catecholamine transporter gene family: molecular cloning and characterization of an L-epinephrine transporter from bullfrog sympathetic ganglia. PMID- 9327881 TI - Positron emission tomography radioligands for dopamine transporters and studies in human and nonhuman primates. PMID- 9327882 TI - Single photon emission computed tomography imaging of dopaminergic function: presynaptic transporter, postsynaptic receptor, and "intrasynaptic" transmitter. PMID- 9327883 TI - Dopamine transporter changes in neuropsychiatric disorders. PMID- 9327884 TI - Molecular and biochemical studies of rat vesicular monoamine transporter. PMID- 9327886 TI - Ligand recognition by the vesicular monoamine transporters. PMID- 9327885 TI - A chimeric vesicular monoamine transporter dissociates sensitivity to tetrabenazine and unsubstituted aromatic amines. PMID- 9327887 TI - Molecular pharmacology of the vesicular monoamine transporter. PMID- 9327888 TI - Ultrastructural localization of the vesicular monoamine transporter 2 in mesolimbic and nigrostriatal dopaminergic neurons. PMID- 9327890 TI - Protein targeting in neurons and endocrine cells. PMID- 9327889 TI - ICA 512, a receptor tyrosine phosphatase-like protein, is concentrated in neurosecretory granule membranes. PMID- 9327891 TI - The vesicular monoamine transporter VMAT2 and vesicular acetylcholine transporter VAChT are sorted to separate vesicle populations in PC12 cells. PMID- 9327892 TI - Recycling of synaptic vesicles. PMID- 9327893 TI - The secretory cocktail of adrenergic large dense-core vesicles: the functional role of the chromogranins. PMID- 9327894 TI - A novel, catecholamine release-inhibitory peptide from chromogranin A: autocrine control of nicotinic cholinergic-stimulated exocytosis. PMID- 9327895 TI - Transcription regulation coupled to calcium and protein kinase signaling systems through TRE- and CRE-like sequences in neuropeptide genes. PMID- 9327897 TI - Catecholamine metabolism. From molecular understanding to clinical diagnosis and treatment. Overview. PMID- 9327896 TI - Imaging of monoaminergic and cholinergic vesicular transporters in the brain. PMID- 9327898 TI - Monoamine oxidase A and B: structure, function, and behavior. PMID- 9327899 TI - Genetic deficiencies of monoamine oxidase enzymes: a key to understanding the function of the enzymes in humans. PMID- 9327900 TI - Biological markers, with special regard to platelet monoamine oxidase (trbc-MAO), for personality and personality disorders. PMID- 9327901 TI - Visualization of monoamine oxidase in human brain. PMID- 9327902 TI - Aliphatic N-methylpropargylamines: monoamine oxidase-B inhibitors and antiapoptotic drugs. PMID- 9327903 TI - Antiapoptotic actions of monoamine oxidase B inhibitors. PMID- 9327904 TI - Therapeutic actions of L-deprenyl in dogs: a model of human brain aging. PMID- 9327905 TI - Apomorphine is a potent radical scavenger and protects cultured pheochromocytoma cells from 6-OHDA and H2O2-induced cell death. PMID- 9327907 TI - X-ray crystallography of catechol O-methyltransferase: perspectives for target based drug development. PMID- 9327906 TI - Catechol O-methyltransferase: characterization of the protein, its gene, and the preclinical pharmacology of COMT inhibitors. PMID- 9327908 TI - Catechol O-methyltransferase inhibition and the treatment of Parkinson's disease. PMID- 9327909 TI - The structure and function of the UDP-glucuronosyltransferase gene family. PMID- 9327910 TI - Catecholamine sulfation in health and disease. PMID- 9327911 TI - Metabolism of endobiotics and xenobiotics by UDP-glucuronosyltransferase. PMID- 9327912 TI - Molecular structure of the carrier responsible for hepatic uptake of catecholamines. PMID- 9327913 TI - Catecholamine uptake and metabolism in the liver. PMID- 9327914 TI - Catecholamine uptake and metabolism in rat lungs. PMID- 9327915 TI - The extraneuronal monoamine transporter exists in human central nervous system glia. PMID- 9327916 TI - Removal of circulating catecholamines by extraneuronal amine transport systems. PMID- 9327917 TI - Catecholamine metabolites in internal jugular plasma: a window into the human brain. PMID- 9327918 TI - Norepinephrine metabolites in plasma as indicators of pharmacological inhibition of monoamine oxidase and catechol O-methyltransferase. PMID- 9327920 TI - Clues to the diagnosis of pheochromocytoma from the differential tissue metabolism of catecholamines. PMID- 9327919 TI - The adrenal gland as a source of dihydroxyphenylalanine and catecholamine metabolites. PMID- 9327921 TI - Catecholamine receptors and signal transduction. Overview. PMID- 9327922 TI - Expression and regulation of alpha 1-adrenergic receptors in human tissues. PMID- 9327923 TI - Alpha 1-adrenoceptor subtypes in the human cardiovascular and urogenital systems. PMID- 9327924 TI - Molecular mechanisms of ligand binding and activation in alpha 1-adrenergic receptors. PMID- 9327925 TI - Expansion of the dopamine D1 receptor gene family: defining molecular, pharmacological, and functional criteria for D1A, D1B, D1C, and D1D receptors. PMID- 9327926 TI - D1/D3 receptor relationships in brain coexpression, coactivation, and coregulation. PMID- 9327927 TI - Mapping the binding-site crevice of the D2 receptor. PMID- 9327928 TI - Mechanisms of beta-adrenergic receptor desensitization and resensitization. PMID- 9327929 TI - Role of beta-arrestins in the intracellular trafficking of G-protein-coupled receptors. PMID- 9327930 TI - G-protein-linked receptors as substrates for tyrosine kinases: cross-talk in signaling. PMID- 9327931 TI - Role of arrestins in G-protein-coupled receptor endocytosis. PMID- 9327932 TI - Subtype-specific regulation of the beta-adrenergic receptors. PMID- 9327933 TI - Structural determinants of alpha 2-adrenergic receptor regulation. PMID- 9327935 TI - Regulation of the D1 dopamine receptor through cAMP-mediated pathways. PMID- 9327934 TI - Regulation of D2 and D3 receptors in transfected cells by agonists and antagonists. PMID- 9327936 TI - Mechanisms for activation of multiple effectors by alpha 1-adrenergic receptors. PMID- 9327937 TI - Signal transduction pathways modulated by D2-like dopamine receptors. PMID- 9327938 TI - Guanosine triphosphatase-activating proteins for heterotrimeric G-proteins. PMID- 9327939 TI - Regulation of the stoichiometry of protein components of the stimulatory adenylyl cyclase cascade. PMID- 9327940 TI - Regulation of mitogen-activated protein kinase pathways by catecholamine receptors. PMID- 9327941 TI - Examination of ligand-induced conformational changes in the beta 2-adrenergic receptor by fluorescence spectroscopy. PMID- 9327942 TI - Relationship between alpha 2-adrenergic receptors and imidazoline/guanidinium receptive sites. PMID- 9327943 TI - Dopamine D4 receptors may alleviate antipsychotic-induced parkinsonism. PMID- 9327944 TI - Binding of typical and atypical antipsychotic drugs to multiple neurotransmitter receptors. PMID- 9327945 TI - Structural and functional characteristics of the dopamine D4 receptor. PMID- 9327946 TI - NGD 94-1: a specific dopamine-4-receptor antagonist. PMID- 9327947 TI - In vivo mutation of the alpha 2A-adrenergic receptor by homologous recombination reveals the role of this receptor subtype in multiple physiological processes. PMID- 9327948 TI - Regulation of fat-cell function by alpha 2-adrenergic receptors. PMID- 9327949 TI - The developmental and physiological consequences of disrupting genes encoding beta 1 and beta 2 adrenoceptors. PMID- 9327950 TI - Myocardial overexpression of adrenergic receptors and receptor kinases. PMID- 9327952 TI - Structure and function of the beta 3 adrenoreceptor. PMID- 9327951 TI - Cardiac G-protein receptor kinase activity: effect of a beta-adrenergic receptor antagonist. PMID- 9327953 TI - Behavioral analysis of multiple D1-like dopamine receptor subtypes: new agents and studies in transgenic mice with D1A receptor knockout. PMID- 9327954 TI - Antisense knockdown of brain dopamine receptors. PMID- 9327956 TI - Regulation of D1 receptor function in spontaneous hypertension. PMID- 9327957 TI - Catecholamines in the periphery. Overview. PMID- 9327955 TI - The physiological role of dopamine D2 receptors. PMID- 9327958 TI - Peripheral catecholaminergic function evaluated by norepinephrine measurements in plasma, extracellular fluid, and lymphocytes, from nerve recordings and cellular responses. PMID- 9327960 TI - Sympathetic microneurography and neurocirculatory function: studies of ventricular arrhythmias in humans. PMID- 9327959 TI - Cardiac microdialysis. PMID- 9327961 TI - Stress as a medical and scientific idea and its implications. PMID- 9327962 TI - Stressor specificity of peripheral catecholaminergic activation. PMID- 9327963 TI - Stressor-specific activation of catecholaminergic systems: implications for stress-related hypothalamic-pituitary-adrenocortical responses. PMID- 9327965 TI - Peripheral modulatory effects of catecholamines in inflammatory and neuropathic pain. PMID- 9327964 TI - Regulation of gene expression of catecholamine biosynthetic enzymes by stress. PMID- 9327966 TI - Brain catecholamine systems in stress. PMID- 9327967 TI - Alpha 2-adrenergic mechanisms of analgesia: strategies for improving their therapeutic window and identification of the novel, potent alpha 2A-adrenergic receptor agonist, S 18616. PMID- 9327968 TI - Cellular transplantation for intractable pain. PMID- 9327969 TI - The role of the sympathetic nervous system in the modulation of immune responses. PMID- 9327970 TI - Catecholamines, catecholamine receptors, cell adhesion molecules, and acute stressor-related changes in cellular immunity. PMID- 9327971 TI - Nerve growth factor and autoimmune diseases: role of tumor necrosis factor-alpha? PMID- 9327972 TI - Multiple transmitter control of catecholamine secretion in rat adrenal medulla. PMID- 9327973 TI - Adrenomedullin in cardiovascular disease. PMID- 9327974 TI - Strychnine, glycine, and adrenomedullary secretion. PMID- 9327975 TI - Catecholamines and neurocardiogenic syncope. PMID- 9327976 TI - Beta-blockers in congestive heart failure: the pharmacology of carvedilol, a vasodilating beta-blocker and antioxidant, and its therapeutic utility in congestive heart failure. PMID- 9327977 TI - Sympathetic cardioneuropathy in dysautonomias. PMID- 9327978 TI - Hypoglycemia-associated autonomic failure in insulin-dependent diabetes mellitus. PMID- 9327979 TI - Mechanisms of the sympathoadrenal response to hypoglycemia. PMID- 9327980 TI - Importance of catecholamines in defense against insulin hypoglycemia in humans. PMID- 9327981 TI - Sympathetic nervous activity and the thermic effect of food in humans. PMID- 9327982 TI - Microdialysis for the assessment of catecholamine-induced lipolysis in adipose and skeletal muscle tissue. PMID- 9327983 TI - Bulbospinal C1-adrenergic neurons: electrophysiological properties in the neonate rat. PMID- 9327984 TI - Catecholamines, opioids, and vagal afferents in the nucleus of the solitary tract. PMID- 9327985 TI - Agmatine: a novel neurotransmitter? AB - Current evidence is consistent with an hypothesis that agmatine meets many criteria for a neurotransmitter-neuromodulator. It is synthesized, stored, and released in brain; is contained in neurons and axon terminals with a heterogeneous distribution; interacts with cell-specific receptors; and elicits biological actions within the central nervous system. Its role in normal brain function, however, has not yet been established, in part because of the absence of agents that selectively affect its biosynthesis or degradation. PMID- 9327986 TI - Central and peripheral norepinephrine kinetics in heart failure, coronary artery disease, and hypertension. PMID- 9327988 TI - Dopamine-mediated gene regulation in the striatum. PMID- 9327987 TI - Catecholamines in the central nervous system. Overview. AB - A goal of this part is to examine the functional impact of catecholaminergic systems on CNS function as it relates to normal and pathological states. The participants achieve their objectives individually by relating their high-quality work on this expansive topic, while maintaining a focus on the functional implications of their findings. Nonetheless, despite the necessarily broad nature of the topics presented, there is a remarkable degree of convergence of information. Several subthemes have emerged as a consequence of considering this work in its entirety. First is the importance of examining neurotransmitter effects, not in isolation, but in terms of interactions with other neurotransmitter systems. History has shown that a limited focus often produces confusing or inconsistent results, which become increasingly clear on consideration of the state of the organism. Second is the importance of examining pharmacological and pathophysiological interactions in light of the anatomy of the system and how developmental influences can alter this relationship. Such very general considerations have been found to provide an essential ingredient in understanding the nature of catecholamine function within this complex system. PMID- 9327989 TI - Dopamine control of gene expression in basal ganglia nuclei: striatal and nonstriatal mechanisms. PMID- 9327990 TI - Dopaminergic regulation of immediate early gene expression in the basal ganglia. PMID- 9327991 TI - Dopamine and calcium signal interactions in the developing striatum: control by kinetics of CREB phosphorylation. PMID- 9327992 TI - The phasic reward signal of primate dopamine neurons. PMID- 9327993 TI - Afferent control of midbrain dopamine neurons: an intracellular perspective. PMID- 9327994 TI - GABAergic control of the firing pattern of substantia nigra dopaminergic neurons. PMID- 9327995 TI - Afferent control of substantia nigra compacta dopamine neurons: anatomical perspective and role of glutamatergic and cholinergic inputs. PMID- 9327996 TI - Dopamine axons in primate prefrontal cortex: specificity of distribution, synaptic targets, and development. PMID- 9327997 TI - The cortical dopamine system: role in memory and cognition. PMID- 9327998 TI - Norepinephrine-dopamine interactions in the prefrontal cortex and the ventral tegmental area: relevance to mental diseases. PMID- 9327999 TI - Dopamine function in the prefrontal cortex. PMID- 9328000 TI - The modulation of corticoaccumbens transmission by limbic afferents and dopamine: a model for the pathophysiology of schizophrenia. PMID- 9328001 TI - Dopamine modulation of responses mediated by excitatory amino acids in the neostriatum. PMID- 9328003 TI - Modulation by dopamine of rat corticostriatal input. PMID- 9328002 TI - The molecular basis of dopamine and glutamate interactions in the striatum. PMID- 9328004 TI - Dopamine, glutamate, and behavioral correlates of striatal neuronal activity. PMID- 9328005 TI - State-related activity, reactivity of locus ceruleus neurons in behaving monkeys. PMID- 9328006 TI - Modulation of forebrain electroencephalographic activity and behavioral state by the locus ceruleus-noradrenergic system: involvement of the medial septal area. PMID- 9328007 TI - New perspectives on the functional organization and postsynaptic influences of the locus ceruleus efferent projection system. PMID- 9328008 TI - Neuromodulation and cognitive performance: recent studies of noradrenergic locus ceruleus neurons in behaving monkeys. PMID- 9328009 TI - Noradrenergic effects on activity of prefrontal cortical neurons in behaving monkeys. PMID- 9328010 TI - Noradrenergic influences on prefrontal cortical cognitive function: opposing actions at postjunctional alpha 1 versus alpha 2-adrenergic receptors. PMID- 9328011 TI - Afferent control of nucleus locus ceruleus: differential regulation by "shell" and "core" inputs. PMID- 9328012 TI - Sensory response of the locus ceruleus: neonatal and adult studies. PMID- 9328013 TI - Noradrenergic modulation of the prefrontal cortex as revealed by electron microscopic immunocytochemistry. PMID- 9328014 TI - Activation of the locus ceruleus brain noradrenergic system during stress: circuitry, consequences, and regulation. PMID- 9328015 TI - Norepinephrine and schizophrenia: a new hypothesis for antipsychotic drug action. PMID- 9328016 TI - Neurochemical responses to lesions of dopaminergic neurons: implications for compensation and neuropathology. PMID- 9328017 TI - Dopamine receptor subtypes as targets for the pharmacotherapy of Parkinson's disease. PMID- 9328018 TI - Free radicals and MPTP-induced selective destruction of substantia nigra compacta neurons. PMID- 9328019 TI - Application of gene therapy for Parkinson's disease: nonhuman primate experience. PMID- 9328020 TI - Prefrontal cortical and hippocampal modulation of dopamine-mediated effects. PMID- 9328022 TI - Interactions between catecholamines and serotonin: relevance to the pharmacology of schizophrenia. PMID- 9328021 TI - Dysregulation of mesoprefrontal dopamine neurons induced by acute and repeated phencyclidine administration in the nonhuman primate: implications for schizophrenia. PMID- 9328023 TI - Novel catecholaminergic systems. Overview. PMID- 9328024 TI - Catecholestrogens in the induction of tumors in the kidney of the Syrian hamster. PMID- 9328025 TI - Biogenesis and inactivation of catecholestrogens. PMID- 9328026 TI - Catecholestrogens as procarcinogens: depurinating adducts and tumor initiation. PMID- 9328027 TI - Role of aromatic hydrocarbons in disclosing how catecholestrogens initiate cancer. PMID- 9328028 TI - Embryo implantation requires estrogen-directed uterine preparation and catecholestrogen-mediated embyronic activation. PMID- 9328029 TI - Extra-adrenal nonneuronal epinephrine and phenylethanolamine-N-methyltransferase. PMID- 9328030 TI - Dopamine and the brain-gut axis. PMID- 9328031 TI - Origin and significance of plasma dihydroxyphenylalanine. PMID- 9328032 TI - Is L-DOPA a neurotransmitter of the primary baroreceptor afferents terminating in the nucleus tractus solitarii of rats? PMID- 9328033 TI - Immunocytochemical evidence of novel catecholamine- or biopterin-related neurons of mammalian brain. PMID- 9328034 TI - Fluorinated dihydroxyphenylserines as potential biological precursors of fluorinated norepinephrines. PMID- 9328035 TI - Nonneuronal dopamine. PMID- 9328036 TI - The renal dopamine system. AB - Intrarenally formed dopamine induced natriuresis by inhibiting the activity of renal tubular Na/KATPase. This effect is mediated via a complex signal network, which includes inhibition of PP1 via the adenylyl cyclase-PKA-DARPP32 pathway and activation of PKC via the PLA2-arachidonic acid-20HETE pathway. The renal dopamine availability is a major determinant of the natriuretic effect of dopamine and is to a large extent modulated by the activity of COMT. The possibility that regulation of dopamine storage and release influences renal dopamine effects should be considered. PMID- 9328037 TI - Renal dopamine production and release in the rat: a microdialysis study. PMID- 9328039 TI - Inductive interactions underlie neural crest formation. AB - The data summarized here indicate that (1) the neural crest can arise by means of inductive interaction between the neural and nonneural ectoderm; (2) initially, progenitor cells are multipotent, having the potential to form multiple ectodermal derivatives (epidermis, neural crest, and neural tube derivatives); and (3) with time, the precursors become progressively restricted to form neural crest derivatives and eventually to individual phenotypes. PMID- 9328038 TI - Development and plasticity. Overview. PMID- 9328040 TI - Lineage commitment and fate of neural crest-derived neurogenic cells. PMID- 9328041 TI - The differentiation of the neurotransmitter phenotypes in chick sympathetic neurons. PMID- 9328042 TI - Developmental regulation of neurotransmitters in sympathetic neurons. PMID- 9328043 TI - Changes in gene expression in adult sympathetic neurons after axonal injury. PMID- 9328044 TI - Ontogeny of vesicular amine transporter expression in the rat: new perspectives on aminergic neuronal and neuroendocrine differentiation. PMID- 9328045 TI - Specification and survival of dopaminergic neurons in the mammalian midbrain. PMID- 9328046 TI - Effects of glial cell line-derived neurotrophic factor on the nigrostriatal dopamine system in rodents and nonhuman primates. PMID- 9328047 TI - Cell body infusions of brain-derived neurotrophic factor increase forebrain dopamine release and serotonin metabolism determined with in vivo microdialysis. PMID- 9328048 TI - Neurotrophin modulation of hippocampal synaptic transmission. PMID- 9328049 TI - Genotype and phenotype in familial dysautonomia. PMID- 9328050 TI - A gene therapy approach for the treatment of amyotrophic lateral sclerosis and Parkinson's disease. PMID- 9328052 TI - Evolution and origin of the diversity of dopamine receptors in vertebrates. PMID- 9328051 TI - Characterization of adrenal chromaffin progenitor cells in mice. PMID- 9328053 TI - Neurogenetics of synaptic transmission in Caenorhabditis elegans. PMID- 9328055 TI - Dopaminergic control of serotonergic neuron development in the grasshopper central nervous system. PMID- 9328054 TI - Decapitated Drosophila: a novel system for the study of biogenic amines. PMID- 9328057 TI - Rapid acquisition of discriminative responding in monkey locus coeruleus neurons. PMID- 9328056 TI - Noradrenergic long-term potentiation in the dentate gyrus. PMID- 9328058 TI - Catecholaminergic contributions to early learning. PMID- 9328059 TI - Interactions between catecholamines and the amygdala in emotional memory: subclinical and clinical evidence. PMID- 9328060 TI - Drug abuse and alcoholism. Overview. PMID- 9328061 TI - Circuits, drugs, and drug addiction. PMID- 9328062 TI - Homologies and differences in the action of drugs of abuse and a conventional reinforcer (food) on dopamine transmission: an interpretative framework of the mechanism of drug dependence. PMID- 9328063 TI - Drug-induced adaptations in catecholamine systems: on the inevitability of sensitization. PMID- 9328064 TI - Neurobiological substrates underlying conditioned effects of cocaine. PMID- 9328065 TI - The rate hypothesis and agonist substitution approaches to cocaine abuse treatment. PMID- 9328066 TI - Drugs of abuse and dopamine cell activity. PMID- 9328067 TI - D1-receptor regulation of synaptic potentials in the ventral tegmental area after chronic drug treatment. PMID- 9328069 TI - Dopamine efflux studies into in vivo actions of psychostimulant drugs. PMID- 9328068 TI - Neuroadaptations in nucleus accumbens neurons resulting from repeated cocaine administration. PMID- 9328070 TI - Psychostimulants and neuropeptide response. PMID- 9328071 TI - Drugs of abuse and striatal gene expression. PMID- 9328072 TI - Coordinated expression of dopamine receptors in neostriatal medium spiny neurons. PMID- 9328073 TI - Dopaminergic genes and substance abuse. PMID- 9328074 TI - Quantitative trait loci: mapping drug and alcohol-related genes. PMID- 9328075 TI - Nuclear memory: gene transcription and behavior. PMID- 9328076 TI - Phosphorylation of dopamine transporters and rapid adaptation to cocaine. PMID- 9328112 TI - Antibodies to Jo-1 and Ro-52: why do they go together? PMID- 9328113 TI - CD28 costimulation and T lymphocyte proliferative responses in HIV-1 infection. AB - To investigate whether defective costimulatory signals could be involved in the loss of T lymphocyte functions during HIV-1 infection, we tested the effect of CD28 costimulation on both T cell receptor/CD3 and HIV-1 antigen-induced proliferative responses. Although CD3-mediated responses significantly decreased with more advanced stages of HIV-1 infection, the ability of potentiating the responses through CD28 costimulation was maintained at all stages and did not differ from that of HIV-1- subjects. When CD28 costimulation was studied in lymphocyte cultures stimulated with HIV-1 gp160 or p24, potentiation was seen only when a significant response was present without additional CD28 triggering, namely in subjects receiving active immunization with recombinant gp160. These results confirm the integrity of the CD28 pathway of costimulation during HIV-1 infection, and suggest that lymphocytes responding to soluble HIV-1 antigen are not deleted in HIV-1-infected patients, but do not receive significant priming during the natural course of the infection. PMID- 9328114 TI - CD8+CD28- T cells in vertically HIV-infected children. AB - To evaluate whether vertical HIV infection interferes with the expression of CD28 on T lymphocytes, 25 HIV-infected children and 29 seroreverted children born to HIV+ mothers were studied. The percentage of CD28- cells among CD8+ T lymphocytes was higher in HIV-infected children than in controls (P < 0.001). In fact, in HIV infected children, this percentage was elevated from the first year of life, while in healthy seroreverted children, the proportion of CD28- cells among CD8+ cells rose progressively with age (r = 0.49; P = 0.008). In HIV+ children, the CD8+ CD28-, but not CD8+ CD28+ cell proportion was significantly correlated with immunological markers of disease progression, such as CD4+ cell loss (r = -0.65; P < 0.001) and the level of in vitro spontaneous lymphocyte apoptosis (r = 0.53; P = 0.03). PMID- 9328115 TI - Characteristics of asymptomatic secondary immune responses to measles virus in late convalescent donors. AB - Among 44 fully protected, late convalescent adults re-exposed to measles, four developed an asymptomatic secondary immune response (SIR) with a significant increase in measles virus (MV)-specific IgG and low IgM. The boosted antibodies were mainly of the IgG1 subclass and reacted with the nucleoprotein and the haemagglutinin protein. About 30 weeks after re-exposure, antibody levels had decreased by 35-50%, suggesting that the booster effect may only be transient. SIR was only found in individuals with a pre-exposure IgG level below a well defined threshold. Antibody levels above this threshold fully protected against SIR. SIR seems to be an 'all or none response' where the magnitude of increase in specific IgG is independent of pre-exposure antibody levels as long as these are below the above threshold. In combination with pre-exposure neutralizing and haemagglutination inhibiting titres, a threshold was defined below which SIR is likely to occur. This may be useful to predict susceptibility to SIR in a given population, since individuals undergoing clinically inapparent SIR are among seropositive subjects, the most likely candidates to support transmission of virus. PMID- 9328116 TI - Interferon-gamma (IFN-gamma) regulates production of IL-10 and IL-12 in human herpesvirus-6 (HHV-6)-infected monocyte/macrophage lineage. AB - To determine whether HHV-6 infection induces expression and production of IL-10 and IL-12 in monocytes/macrophages, and to explore the influence of IFN-gamma on cytokine production in HHV-6-infected cells, expression and production of IL-10 and IL-12 were evaluated through reverse transcription-polymerase chain reaction (RT-PCR) and sandwich ELISA. HHV-6 infection induced the expression and the production of IL-10 and IL-12 in monocytes and THP-1 cells. Kinetic study showed that the expression of IL-12 mRNA decreased with accumulation of IL-10 mRNA. Expression and production of IL-12 were markedly increased when anti-human IL-10 MoAbs were added to the cultures, implying that endogenous IL-10 induced by HHV-6 inhibited IL-12 production. Addition of increasing concentrations of IFN-gamma to the cultures of HHV-6-infected cells enhanced the expression of IL-12 gene, while the accumulation of IL-10 mRNA was down-regulated. Determination of protein levels of IL-10 and IL-12 by ELISA also showed that IFN-gamma increased IL-12 and decreased IL-10 production. These results suggest that IFN-gamma regulates the production of IL-10 and IL-12 at transcriptional level mainly through inhibiting endogenous IL-10 production in HHV-6-infected monocyte/macrophage lineage. PMID- 9328118 TI - Antagonistic effects of IL-4 and interferon-gamma (IFN-gamma) on inducible nitric oxide synthase expression in bovine macrophages exposed to gram-positive bacteria. AB - Cytokine-mediated modulation of nitric oxide (NO) production by bacteria stimulated bovine macrophages was studied. When Salmonella dublin, as a prototypic gram-negative organism, was used, NO generation was barely enhanced by recombinant bovine and ovine IFN-gamma, but was suppressed by IL-4. Salmonella dublin-induced NO generation was not influenced by a panel of nine other cytokines. The panel included IL-1, tumour necrosis factor (TNF) and IFN-alpha, which are active in a similar mouse macrophage model. The tested cytokines were either homologous or known to interact with bovine cytokine receptors. Recombinant bovine and ovine IFN-gamma were the only cytokines which strongly enhanced NO synthesis by macrophages exposed to the gram-positive organism, Listeria monocytogenes. Listeria-induced NO generation was strongly suppressed by recombinant human and bovine IL-4, but not by IL-10 and transforming-growth factor-beta. Thus, two cytokines characterizing a Th1 and a Th2 response up- and down-regulate, respectively, bacteria-induced NO generation in bovine macrophages, whereas nine other cytokines had little activity in this regard. This modulation was reflected in changes in the steady state levels of mRNA coding for inducible nitric oxide synthase. Combinations of IFN-gamma and IL-4 suggested that the relative proportion of these cytokines determined whether bacteria-induced NO generation was up- or down-regulated. At saturating IL-4 concentrations, stimulation of bacteria-induced NO generation in macrophages by T cell supernatants was solely dependent on IFN-gamma. This was shown by antibody neutralization experiments and by a close correlation between the capacity of supernatants to stimulate NO generation and the IFN-gamma content, as determined by immunoassay. PMID- 9328117 TI - The production of IL-8 in cerebrospinal fluid in aseptic meningitis of children. AB - Neutrophils accumulate initially in the cerebrospinal fluid (CSF) of aseptic meningitis, perhaps because of increased levels of granulocyte colony-stimulating factor (G-CSF), macrophage inflammatory protein-1alpha (MIP-1alpha), and IL-8 in the subarachnoid space. We studied levels of these cytokines in children with aseptic meningitis using ELISA. When meningeal symptoms existed, IL-8 levels (1399 +/- 1600 ng/l, n = 32) in the CSF were significantly higher than those either after meningeal symptoms disappeared (61 +/- 56 ng/l, n = 18) or in controls (44 +/- 63 ng/l, n = 27) (P < 0.0001). High levels of IL-8 on admission dropped sequentially. Significant correlations were found between IL-8 levels and either neutrophil counts (r = 0.612), G-CSF levels (r = 0.873) or MIP-1alpha levels (r = 0.623) in the CSF of the affected patients (P < 0.0001). IL-8 values in serum were lower than in the corresponding CSF samples from all individuals with meningeal symptoms. The IL-8 mRNA was detectable by reverse-transcribed polymerase chain reaction (PCR)-assisted amplification in fresh leucocytes from the CSF, but not from the peripheral blood of a healthy volunteer. The culture of CSF mononuclear cells produced high levels of IL-8 (approximately 2750 ng/l). These data indicate that IL-8 levels rise transiently at the initial stage of aseptic meningitis, and that mononuclear cells that migrate into the CSF are a cellular source of this chemokine. We suppose that IL-8, in addition to G-CSF and MIP-1alpha, contribute to the localized neutrophil accumulation during the disease. PMID- 9328119 TI - Identification and characterization of a DR4-restricted T cell epitope within chlamydia heat shock protein 60. AB - An epitope within the 60 kD Chlamydia trachomatis heat shock protein (hsp) 60, recognized by a HLA-DRB1*0401-restricted T cell clone from a reactive arthritis patient, has been characterized. Stimulatory peptides contained a nine amino acid sequence (residues 38-46) predicted by algorithm to confer strong binding to DRB1*0401, with valine in the P1 position. The overall length of the peptide was critical for efficient recognition; peptides with at least one residue N-terminal to the putative P1 position were markedly more stimulatory than a peptide whose N terminal is the P1 valine. Optimal responses were seen with 14mer peptides having two to three amino acids N- and C-terminal to the core 9mer. The sequence of the defined epitope is identical in hsp60 from both C. trachomatis and C. pneumoniae. Since the latter is a common respiratory pathogen, patients infected with C. trachomatis may already be primed for responses to hsp60 by prior infection with C. pneumoniae. Such secondary responses are important in the pathogenesis of chlamydia-induced inflammatory diseases such as trachoma. Priming by infection with enteric organisms was considered because of the similarity of the epitope sequence in Escherichia coli hsp60. However, although an E. coli-related peptide was recognized, intact E. coli hsp60 was not, suggesting that the epitope is cryptic in E. coli hsp60. Human hsp60 has six amino acid differences from chlamydial hsp60 in the epitope sequence and was not recognized. Thus cross reactive recognition of self hsp60 could not be implicated in the pathogenesis of chlamydia-induced reactive arthritis in this patient. PMID- 9328120 TI - Fractionation of mycobacterial integral membrane proteins by continuous elution SDS-PAGE reveals the immunodominance of low molecular weight subunits for human T cells. AB - Integral membrane proteins (IMP) represent a serologically distinct class of mycobacterial antigens which are potent stimulators of human T cells (Mehrotra et al., Clin Exp Immunol 1995; 102:626). The range of IMP from Mycobacterium fortuitum was resolved by continuous elution SDS-PAGE to recover 31 discrete fractions covering bands up to approximately 58 kD. The fractions, after removal of SDS, were subjected to human T cell proliferation assays for the identification of immunodominant molecule(s). A low molecular weight (<20kD) fraction was able to stimulate T cells from 11 out of 12 donors comprising mainly tuberculoid leprosy patients. The described protocol is well suited to situations where large quantities of antigenic protein mixtures must be processed in order to get the purified molecules/fractions in amounts required for immunoepidemiological studies. PMID- 9328121 TI - Impairment of natural killer (NK) cytotoxic activity in hepatitis C virus (HCV) infection. AB - In the present study, we evaluated the NK cell cytotoxic activity in a group of HCV-infected individuals. Although the number of NK cells present in the peripheral blood of the HCV-infected patients was comparable to non-infected individuals, spontaneous NK cytotoxicity was four-fold lower (P < 0.001) than in normal donors. This functional impairment was not overcome by depletion of adherent or B cells, and it was partially restored by short-term (18 h) stimulation with IL-2. However, long-term stimulation (72 h) with this lymphokine induced activated killer cell (LAK) activity comparable to normal controls. The reduction in NK cytotoxic response does not seem to be due to soluble suppressive factors, since incubation of normal peripheral blood mononuclear cells (PBMC) with infectious HCV serums for a 4-h period does not affect NK spontaneous cytotoxic activity. Successful in vitro infection of PBMC with HCV infectious serum also resulted in an impairment of NK cytotoxicity, suggesting that altered NK function is associated with HCV infection and may be responsible, at least in part, for the chronicity of the infection. PMID- 9328122 TI - Serum levels of IL-10, IL-15 and soluble tumour necrosis factor-alpha (TNF-alpha) receptors in type C chronic liver disease. AB - We previously reported that the number of TNF-alpha-producing cells was increased in the liver of patients with type C chronic liver disease. To understand further the pathophysiology of this change, we examined serum levels of two soluble TNF receptors, TNF-alphaRI (p55) and -alphaRII (p75), and IL-10, all of which act as TNF-alpha buffer, and IL-15, a novel cytokine sharing many immunological activities with IL-2, using ELISA methods. We studied control individuals and patients with type C chronic liver disease, including asymptomatic hepatitis C virus (HCV) carriers with persistently normal serum ALT values, and those with chronic hepatitis (CH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Both types of sTNF-alphaR closely correlated with disease progression. Patients with LC and HCC had significantly elevated levels for sTNF-alphaRII compared with the other patient groups and controls. Serum IL-10 levels were significantly greater in all chronic liver disease groups than in controls. With respect to IL 15, the values were high in CH, LC and HCC compared with those of controls. Notably, HCC patients showed highest values for both IL-10 and IL-15, with significant differences from the other patient groups. Serial determinations revealed that interferon (IFN) treatment for CH patients resulted in the suppression of circulating IL-10 and IL-15 levels along with decrease in serum aminotransferase values. Both cytokines remained at decreased levels after cessation of therapy in patients who went into clinical and virological remission. On the other hand, treatment did not affect serum levels of sTNF alphaRs. These findings indicate that serum levels of these molecules correlated with disease progress in chronic HCV infection, and that IL-10 and IL-15 may reflect the degree of inflammation in the liver. It is also suggested that both cytokines may be related to the development of HCC. PMID- 9328123 TI - Density of gamma/delta+ T cells in the jejunal epithelium of patients with coeliac disease and dermatitis herpetiformis is increased with age. AB - Increased density of gamma/delta T cell receptor (TCR)+ intraepithelial lymphocytes is the only characteristic in the jejunum of patients with coeliac disease and dermatitis herpetiformis which is not normalized on a gluten-free diet. We explored the age-dependent changes in intraepithelial gamma/delta and alpha/beta TCR+ cells from 137 biopsies from patients with coeliac disease and dermatitis herpetiformis and from controls. Biopsy specimens from 100 patients with coeliac disease and dermatitis herpetiformis and from 37 controls were studied with an immunohistochemical method using MoAbs to T cell receptors and peroxidase staining. An increase in the density of intraepithelial gamma/delta T cells above the mean +2 s.d. of the density in controls was present in 97 of 100 specimens from patients with coeliac disease and dermatitis herpetiformis. The density of gamma/delta+ cells of patients with coeliac disease and dermatitis herpetiformis on a normal gluten-containing diet showed a positive correlation with age (r = 0.45, P < 0.0001). In controls, the density of gamma/delta+ cells remained low throughout the age-range studies, from age 0.6-57 years. In controls, alpha/beta+ cells increased with age (r = 0.57, P < 0.001). The increase in density of intraepithelial lymphocytes with age is in agreement with their thymus-independent character and local proliferation. PMID- 9328125 TI - Hypogammaglobulinaemia occurs in Fas-deficient MRL-lpr mice following deletion of MHC class II molecules. AB - Fas (CD95)-mediated apoptosis in B and T cells is deficient in both human autoimmune lymphoproliferative syndrome and in MRL-lpr mice, a model for systemic lupus erythematosis (SLE). Autoimmune disease in these mice is associated with polyclonal B cell activation, increased serum immunoglobulin and autoantibodies. In non-autoimmune mice MHC class II is not required for normal serum immunoglobulin expression, and previously we have shown using MHC class II deficient MRL-lpr mice (MRL-lpr Ab-/-) that generation of specific antibodies to DNA requires MHC class II-directed T cell help. In contrast, in the present study we demonstrate that MRL-lpr Ab-/- mice also have a profound reduction of total serum immunoglobulin levels, suggesting abnormal polyclonal regulation of B cells by MHC class II-directed T cells occurs in the autoimmune MRL-lpr strain. This abrogation of immunoglobulin production does not occur in MHC class II-deficient non-obese diabetic (NOD) mice, nor in MHC class I-deficient NOD or MRL-lpr mice. Reduced immunoglobulin levels in MRL-lpr Ab-/- mice were not due to a lack of B cells or to an increased loss of circulating immunoglobulin, but were associated with reduced numbers of surface IgG-positive B cells. These results define a general abnormal regulation of B cells in MRL-lpr mice through a process requiring MHC class II, and suggest that Fas deficiency may allow expansion of totally T-dependent B cells. PMID- 9328124 TI - Chronic sinusitis refractory to standard management in patients with humoral immunodeficiencies. AB - Chronic refractory sinusitis is a common feature in patients with primary immunodeficiencies. The efficacy of standard therapeutic strategies is questionable. In an open trial we evaluated the efficacy of azithromycin, N acetylcysteine and topical intranasal beclomethasone (100 microg twice daily for 6 weeks) in 16 patients with primary immunodeficiencies (median age 13.5 years, range 5-32 years). All patients suffered from chronic sinusitis despite regular immunoglobulin replacement therapy every 3 weeks. Magnetic resonance imaging (MRI) scans were performed before and after 6 weeks of treatment to evaluate morphological changes in the paranasal sinuses. Nasal swabs and washings were taken for microbial analysis and measurement of inflammatory mediators (IL-8, tumour necrosis factor-alpha (TNF-alpha), eosinophilic cationic protein (ECP)) before and post therapy. Inflammatory mediators in nasal secretions were significantly elevated in patients: IL-8 median 2436 pg/ml (range 441-5435 pg/ml), TNF-alpha 37.3 pg/ml (3.75-524 pg/ml) and ECP 33 ng/ml (1.5-250 ng/ml) versus age-matched healthy controls: IL-8 median 212 pg/ml (99-825 pg/ml), TNF alpha 3.77 pg/ml (2.8-10.2 pg/ml) and ECP 1.5 ng/ml (1.5-14.8 ng/ml) (P < 0.0001). Inflammation of the maxillary sinuses was confirmed by MRI scans in all patients, additionally infection of the ethmoidal and frontal sinuses was recorded in five patients. Bacterial growth appeared in 11 out of 16 cultures. In spite of therapy, no improvement in sinal inflammation visualized by MRI was achieved. Moreover, no significant decrease in pathogens and levels of inflammatory mediators could be detected (IL-8 1141 pg/ml, 426-4556 pg/ml; TNF alpha 13.9 pg/ml, 4.1-291.6 pg/ml; ECP 32.3 ng/ml, 3.7-58.4 ng/ml). Our results demonstrate that conventional management of sinusitis is of little benefit in patients with chronic refractory sinusitis with an underlying immunodeficiency. More studies are needed to test antibiotic regimens, probably combined with surgical drainage and anti-inflammatory agents. PMID- 9328126 TI - Anti-Ro fine specificity defined by multiple antigenic peptides identifies components of tertiary epitopes. AB - Anti-Ro (or SSA) is a clinically important autoantibody that is found in 25-40% of patients with systemic lupus erythematosus as well as an even greater proportion of patients with Sjogren's syndrome or subacute cutaneous lupus. We have studied the binding of anti-Ro sera to multiple antigenic peptides constructed from the sequence of the 60-kD Ro molecule. The results demonstrate that sera bind these peptides in solid-phase assay. Surprisingly, some of these peptides also form a precipitin line in double immunodiffusion with anti-Ro sera. Formation of lines of identity in double immunodiffusion as well as absorption studies indicate that peptides distant in the primary amino acid sequence and without shared sequence are bound by the same antibody. In addition, data from surface plasmon resonance demonstrate that peptides identified in this manner have protein-protein interactions. Thus, these techniques may identify the components of conformational epitopes. PMID- 9328127 TI - Autoantibody reactive with the human general transcription factor TFIIF in sera from patients with autoimmune disorders. AB - Transcription factor (TF) IIF, a heteromeric protein composed of two subunits, RAP30 and RAP74, is required for both specific initiation and elongation of mRNA synthesis by RNA polymerase II. We have identified high titre of specific autoantibodies against the RAP74 subunit of TFIIF in sera from patients with systemic lupus erythematosus (SLE) as well as those with rheumatoid arthritis (RA), periarteritis nodosa (PN), Sjogren's syndrome (SS), dermatomyositis (DM), and mixed connective tissue disease (MCTD) by Western blot or immunoprecipitation. The epitopes recognized by autoantibodies were shown to be preferentially clustered at the central charged region. Anti-RAP74 autoantibody was shown to suppress the activity of TFIIF-stimulated elongation of mRNA synthesis by RNA polymerase II. It is concluded that some patients with autoimmune disorders develop specific autoantibodies against the RAP74 subunit of TFIIF. PMID- 9328128 TI - Immunoprecipitation of melanogenic enzyme autoantigens with vitiligo sera: evidence for cross-reactive autoantibodies to tyrosinase and tyrosinase-related protein-2 (TRP-2). AB - In the present study we describe the detection of TRP-2 antibodies in vitiligo patients using in vitro 35S-labelled human TRP-2 in a radioimmunoassay. Of 53 vitiligo sera examined in the assay, three (5 9%) were found to be positive for TRP-2 antibodies. In contrast, 20 control sera, sera from 10 patients with Hashimoto's thyroiditis and sera from 10 patients with Graves' disease were all negative. All three patients positive for TRP-2 antibodies (mean age 54 years, age range 50-63 years) had had vitiligo of the symmetrical type for more than 1 year and all of them also had an associated autoimmune disorder: Graves' disease in one and autoimmune hypothyroidism in two. In addition, antibodies to the melanogenic enzyme tyrosinase were present in their serum. To examine any immunological cross-reactivity between TRP-2 and tyrosinase, the three vitiligo sera positive for TRP-2 antibodies were preabsorbed with COS-7 cell extract containing either expressed TRP-2 or tyrosinase, and subsequently used in the radioimmunoassay. These absorption studies indicated that preincubation with both proteins inhibited the immunoreactivity of the positive sera in the immunoassay using in vitro translated 35S-TRP-2. This antibody cross-reactivity suggests the humoral response to the two melanogenic enzymes in these patients may not be entirely independent. PMID- 9328129 TI - Phenotype, cytokine production and cytolytic capacity of fresh (uncultured) tumour-infiltrating T lymphocytes in human renal cell carcinoma. AB - We investigated the phenotype and functional capacities of tumour-infiltrating lymphocytes (TIL), freshly isolated from primary renal cell carcinoma (RCC) specimens (n = 20). Three-colour flow cytometry immunophenotyping revealed that RCC TIL consist mainly of CD3+ T cells, with a clear predominance of CD4- CD8+ over CD4+ CD8- T cells, and a marked population of CD4+ CD8+ T cells. Natural killer (NK) cells were also strongly represented (> 25% in 15 of 20 tumour samples), while B cells constituted a minor TIL subset (< 5% in 18 of 20 tumour samples). More importantly, the T and NK cells within the tumour displayed a significantly higher expression of the early activation marker CD69 than their counterparts in adjacent normal renal tissue and in peripheral blood. Expression of CD54 and of HLA-DR was also elevated on CD3+ TIL, and HLA-DR expression was further vigorously up-regulated following ex vivo stimulation with anti-CD3, all suggesting enhanced immune activity within the tumour microenvironment. CD3+ CD4+ TIL displayed a normal capacity to up-regulate CD25 expression and to secrete both Th1-type (IL-2, tumour necrosis factor-alpha (TNF-alpha) and interferon gamma (IFN-gamma)) and Th2-type (IL-4, IL-5 and IL-10) cytokines upon triggering with anti-CD3. Furthermore, cytokine production was susceptible to modulation by CD28 costimulation. CD3+ CD8+ TIL, on the other hand, consistently demonstrated a poor up-regulation of CD25 upon triggering with anti-CD3, and displayed poor ex vivo cytolytic activity in an anti-CD3-redirected 4-h cytotoxicity assay against murine P815 cells. Collectively, our findings indicate that the CD3+ CD4+ TIL in RCC have normal functional capacities, whereas the proportionally major CD3+ CD8+ TIL are functionally impaired. The relevance of these findings to the in vivo local immune response in RCC is discussed. PMID- 9328130 TI - C1q, a subunit of the first component of complement, enhances antibody-mediated apoptosis of cultured rat glomerular mesangial cells. AB - We have shown previously that IgG2a anti-Thy-1 MoAb (ER4G) induces apoptosis of rat mesangial cells (GMC) in vitro. Since the classical complement pathway plays an essential role in Thy-1 nephritis, we analysed whether C1q, a subunit of the first component of complement, enhances the ER4G-mediated apoptosis of rat GMC. Two different subclasses of anti-Thy-1 MoAb, ER4G (IgG2a) and ER14 (IgG1), were used. It was established that ER4G binds C1q efficiently, while ER14 reacts poorly with C1q. For the experiments of apoptosis, quiescent rat GMC were exposed for 1 h at 37 degrees C to a fixed concentration of anti-Thy-1 MoAb and incubated further for 16 h at 37 degrees C in the presence or absence of C1q. GMC exposed to medium (M-GMC) followed by incubation of the cells with medium alone was used as controls. Apoptosis was assessed by morphological studies and quantitative analysis on FACS using FITC-annexin V (the annexin V methods) or bicolour FACS analysis using FITC-annexin V and propidium iodide (the annexin V/PI method). With the annexin V method, M-GMC revealed 9.4 +/- 1.4% apoptosis. C1q had only marginal effects on apoptosis of M-GMC. GMC exposed to ER4G (ER4G-GMC) and further incubated with medium in the absence of C1q resulted in 25.7 +/- 5.7% apoptosis (P < 0.01 relative to control). Incubation of ER4G-GMC together with 100 microg/ml of C1q significantly increased GMC-apoptosis up to 39.4 +/- 4.9% (P < 0.01 relative to ER4G-GMC incubated in the absence of C1q). This enhancing effect of C1q on apoptosis of ER4G-GMC was time- and dose-dependent. In contrast, C1q did not significantly alter the apoptosis of either GMC exposed to ER14 (ER14 GMC) or to F(ab')2-ER4G (F(ab')2-ER4G-GMC), while ER14-GMC or F(ab')2-ER4G-GMC incubated with medium resulted in significant apoptosis compared with control. These results were supported by morphological studies and bicolour FACS analyses in time course experiments using the annexin V/PI method. The effect of C1q is dependent on the presence of intact C1q-containing globular heads and does not occur with collagen-like fragments of C1q. Furthermore, incubation of ER4G-GMC with anti-mouse K-chain antibodies also increased ER4G-mediated GMC-apoptosis. These results indicate for the first time that C1q enhances antibody-mediated apoptosis of rat GMC in vitro, presumably by its binding to ER4G and probably by additional cross-linking of Thy-1 on the surface of GMC. PMID- 9328131 TI - Differential effects of cytomegalovirus infection on complement synthesis by human mesangial cells. AB - Viruses may be eliminated by the host immune system via complement-mediated lysis of infected cells. Previously, we have demonstrated that the synthesis of complement factor B by renal mesangial cells (MC) is enhanced by interferon-alpha (IFN-alpha), -beta and -gamma. In the present study we investigate the effect of human cytomegalovirus (HCMV) infection on the production of complement factors by MC. The production of factor B, C2, C4 and factor H by mock-infected MC was 0.2 +/- 0.4, 3.9 +/- 6.8, 1.7 +/- 0.8 and 149 +/- 36 ng/10(6) cells per 72 h, respectively. In HCMV-infected MC cultures an induction of both factor B and C2 protein synthesis was observed up to 2.2 +/- 1.1 and 156 +/- 74 ng/10(6) cells per 72 h, respectively. The synthesis of C4 and factor H of 2.9 +/- 2.0 and 146 +/- 31 ng/10(6) cells, respectively, was not altered significantly. By Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis it was demonstrated that factor B and C2 mRNA expression were up-regulated in HCMV infected cell cultures, whereas the levels of C4 and factor H mRNA were not changed. When MC cultures were inoculated with heat- or UV-inactivated HCMV no enhancement of factor B mRNA expression was observed. The enhanced expression was not blocked by phosphono acetic acid (PAA), suggesting that expression of the HCMV immediate early or early genes is sufficient to induce complement synthesis. We conclude that infection of MC cultures with HCMV selectively induces complement C2 and factor B production, probably mediated by interferons. PMID- 9328132 TI - IL-13 over-expression in skin is not confined to IgE-mediated skin inflammation. AB - IL-13 is produced by T cells and, like IL-4, it can induce the production of IgE and IgG4. In order to investigate if IL-13 is a specific marker for atopic dermatitis (AD), IL-13 gene expression was analysed in chronic lichenified lesions and non-lesional skin of patients with AD, in involved and non-involved skin of patients with psoriasis, in positive tuberculin reactions in non-atopics, and in the skin of healthy control subjects. Patients with AD (n = 9) showed sensitization to common air-borne allergens (positive Phadiatop) and had total serum IgE values in the range from 10 to 4800 kU/l (median 170 kU/l). Competitive reverse transcription-polymerase chain reaction (RT-PCR) was used to assess IL-13 gene expression in skin biopsy specimens. IL-13 gene expression was markedly higher in chronic lichenified lesions of patients with AD (P < 0.01), and in the positive tuberculin reactions (P < 0.01; n = 12) than in skin from healthy control subjects (n = 10). However, there was no significant difference in IL-13 gene expression in the skin of patients with psoriasis (n = 10) and that of healthy control subjects. The dermal cell infiltrates were larger and the relative amount of CD3+ and CD4+ cells in these infiltrates was higher in the skin of subjects with a positive tuberculin reaction than in lichenified AD skin. However, these differences were not reflected in differences in IL-13 gene expression. Different triggers of IL-13 gene expression may influence the diverse patterns of inflammation seen in different inflammatory skin disorders. PMID- 9328133 TI - Elevated serum levels of soluble CD30 in patients with atopic dermatitis (AD). AB - The immunopathology of AD is still unclear, but evidence for an immune response polarized towards Th2 activity has been provided. The CD30 molecule belongs to the tumour necrosis factor (TNF) receptor family and is expressed on activated T cells with a sustained expression in Th2 cells. This molecule also exists in a soluble form (sCD30). Elevated serum levels of sCD30 have been found in patients with Hodgkin's disease, chronic hepatitis B infection and HIV infection. Studies were undertaken to compare the serum levels of sCD30 in patients with AD (n=49) and healthy non-atopic controls (n=94). The presence of sCD30 was analysed with ELISA. A significantly higher concentration of sCD30 was noted in AD patients, median sCD30 level 29 U/ml (range 1-708 U/ml), compared with healthy non-atopic controls (P<0.001), where the median level was 11 U/ml with a range of 1-1042 U/ml. No correlation was found between sCD30 levels and total serum IgE, or between the AD patients' SCORAD values and concentration of sCD30. sCD30 levels were also analysed in 20 AD patients, which during ketoconazole treatment had improved their clinical scores and reduced their serum IgE and eosinophil cationic protein levels. However, no significant decrease in sCD30 levels was noted after treatment. The results show that patients with AD have elevated levels of sCD30, but without correlation to total serum IgE or disease activity. PMID- 9328134 TI - Expression of human-Torpedo hybrid acetylcholine receptor (AChR) for analysing the subunit specificity of antibodies in sera from patients with myasthenia gravis (MG). AB - The nicotinic AChR, a pentamer composed of alpha2betagamma(or epsilon)delta subunits, is the autoantigen in the human autoimmune disease MG. Anti-AChR antibodies in MG sera bind mainly to conformational epitopes, therefore determination of their specificities requires the use of intact AChR. Indirect antibody competition studies have suggested that most MG antibodies are inhibited from binding to AChR by MoAb to the main immunogenic region (MIR) on the alpha subunits. More recently, based on the knowledge that MG antibodies show little detectable cross-reaction with Torpedo AChR, we have shown, using mouse-Torpedo hybrid AChR, that most MG antibodies that detectably cross-react with the mouse AChR bind to the alpha-subunit. To analyse the whole anti-AChR antibody repertoire in MG sera, we expressed on stably transfected fibroblasts a novel human alpha+ Torpedo betagammadelta AChR and compared the antibody titres against human, Torpedo, and the hybrid AChR. Direct information was provided for the subunit specificity of several MoAbs and sera from 50 MG patients. On average, at least 48% of the anti-AChR antibodies in the sera were directed against the alpha subunit. Interestingly, the anti-alpha-subunit antibodies predominated in low titre (0.6-7.4 nM) but not in high titre (10-386 nM) sera, where they comprised on average 68% versus 23% of the antibodies, respectively. Finally, the directly determined anti-alpha-subunit antibodies and the anti-MIR antibodies defined by antibody competition were significantly correlated, thus suggesting that at least a significant fraction of the anti-MIR antibodies in MG sera bind to the alpha subunit. PMID- 9328135 TI - Characterization of fertilization-blocking monoclonal antibody 1G12 with human sperm-immobilizing activity. AB - A mouse hybridoma (1G12) producing sperm-immobilizing MoAb to human sperm was established and characterized in order to study the antigens relevant to sperm immobilization by antibodies. MoAb 1G12 had strong sperm-immobilizing and agglutinating activities and also showed a fertilization-blocking activity on in vitro fertilization tests. The antibody absorption experiments showed that MoAb 1G12 reacted not only to ejaculated sperm but also human seminal plasma, suggesting that the corresponding antigen might be a sperm coating antigen. The MoAb also reacted with peripheral blood lymphocytes. In histochemical studies, the epithelia of corpus epididymis were most strongly stained. Ejaculated sperm were stained with a granular pattern for their entire surface by immunofluorescence. MoAb 1G12 recognized polymorphic glycoproteins of 15-25 kD in the ejaculated sperm extract in Western blot analysis. After deglycosilation of the sperm extract, only a single staining band of under 15 kD was detected by MoAb 1G12. This suggests that the antigen epitope recognized by MoAb 1G12 might be a peptide of the core portion of the glycoprotein. MoAb 1G12 might be a useful tool for studying the mechanism of egg-sperm interaction, and also be applied to identifying the corresponding antigen by using gene technology. PMID- 9328137 TI - Sex and parity modulate cytokine production during murine ageing. AB - We have previously shown that physiological hormone differences related to pregnancy or sex affect the age-related distribution of mononuclear cell populations during murine ageing. To determine whether such changes are involved in the age-related changes in functions of T cells, we examined the secretion of major T cell immunoregulatory cytokines (IL-2, IL-4, interferon-gamma (IFN gamma), IL-3, IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF)) of in vitro concanavalin A-activated spleen cells of C57B1/6 mice. The study included multiparous and virgin females and males at 2, 8, 15 and 23 months of age. Short-term effects of parity (8 months) were evidenced by the decrease of IFN-gamma and the preserved IL-2 production in multiparous females (8 months), while IFN-gamma was unchanged and IL-2 decreased in virgin mice. The increase in IL-4 production appeared earlier in multiparous females (15 months) than in virgin mice (23 months). The increase in IL-4/IFN-gamma and IL-4/IL-2 ratios at 8 and 15 months, respectively, in multiparous females, suggests that pregnancy modifies the Th1/Th2 equilibrium. In late adulthood (15 months), IL-6 and GM-CSF production was higher in multiparous females than in virgin males or females. Sex differences were also noticed: IFN-gamma secretion capacity was lower in males than in females during ageing. This study underlines that the onset, magnitude and kinetics of the age-related changes in cytokine production are parity- and sex-dependent. These changes probably influence the incidence of age-related diseases and may explain the greater longevity of females. PMID- 9328136 TI - High incidence of autoimmune dacryoadenitis in male non-obese diabetic (NOD) mice depending on sex steroid. AB - The NOD mouse develops spontaneous autoimmune lesions in the lacrimal and salivary glands, besides a well characterized T cell-mediated autoimmune pancreatic beta cell lesion. We report unique pathological findings developed in the lacrimal glands as an autoimmune dacryoadenitis of NOD mice in contrast to those found in the salivary glands and pancreas. A high incidence of autoimmune lesions in the lacrimal glands was observed exclusively in male NOD mice at any age. Histology of autoimmune dacryoadenitis in male NOD mice showed severe destructive changes compared with those observed previously as an autoimmune lesion in the lacrimal glands. Castration in male NOD mice significantly decreased the incidence of autoimmune dacryoadenitis, and testosterone treatment with castration also increased the incidence of autoimmune lesions. Oestrogen treatment with castration did not increase the incidence, but tamoxifen treatment without castration significantly increased the incidence of autoimmune dacryoadenitis in NOD mice. In addition, we detected up-regulation of local cytokine genes (IL-1beta, tumour necrosis factor-alpha (TNF-alpha), IL-2, interferon-gamma (IFN-gamma), IL-6, IL-10, and IL-12 p40) during the course of autoimmune dacryoadenitis. These data suggest that in spontaneous autoimmune dacryoadenitis of male NOD mice there may be an intimate relationship with sex steroids, particularly testosterone, in the development and progression of autoimmune lesions, and autoreactive Th1 cells secrete up-regulated cytokine genes, including IL-10 and IL-12. PMID- 9328138 TI - Regulation of cytokine gene expression by adjuvants in vivo. AB - Antibody isotype affects biological activity of the antibodies and therefore should be considered in prevention of disease by vaccination. In previous reports, we demonstrated that adjuvants affect the antibody isotype switching process and favour the production of certain isotypes. The present study extends these findings and shows fundamental differences in the cytokine induction pattern according to the adjuvant used. Cytokine mRNA levels were determined by in situ RNA-RNA hybridization performed on splenocytes isolated from mice injected with different adjuvants. The results revealed that Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA), Al(OH)3 and QuilA administration results in a type-2 (humoral) response, increasing IL-4, IL-5 and IL-13 gene expression, while poly I:C exhibits a type-1 (cell-mediated) response, increasing the production of interferon-gamma (IFN-gamma), IL-2 and IL-6 mRNA. Finally, BeSO4 and poly A:U augment IL-5 and IL-6 mRNA production, while lipopolysaccharide (LPS) and LiCl augment IL-6 and tumour necrosis factor-alpha (TNF-alpha) mRNA production. Also, the adjuvants appear capable of overcoming the inherent IL-2/IFN-gamma and IL-4 dichotomy of C57B1/6 and BALB/c mice, respectively, in response to cellular antigens such as Leishmania and herpes simplex virus (HSV). The overall data suggest that adjuvants direct the isotype switching process via induction of certain cytokines, a finding that can be useful in selection of the most efficient isotype of protective antibodies for disease prevention by vaccination. PMID- 9328139 TI - Complexity of expression of the intermediate filaments of six new human ovarian carcinoma cell lines: new expression of cytokeratin 20. AB - Six permanent human ovarian carcinoma cell lines (OVISE, OVTOKO, OVMANA and OVSAYO from clear cell adenocarcinoma, and OVSAHO and OVKATE from serous papillary adenocarcinoma) were established from solid tumours. The cell lines have been in culture for 5-8 years, the passage number varying from 62 to 246. Immunohistochemical analysis has shown that five of the six cell lines express at least six cytokeratin (CK) polypeptides. OVISE and OVSAYO expressed CKs 6, 7, 8, 18, 19 and 15 and/or 16. OVTOKO was positive for CKs 7, 8, 18, 19 and 15 and/or 16. OVSAHO expressed CKs 6, 7, 8, 14, 18, 19 and 15 and/or 16. OVMANA expressed CKs 6, 7, 8, 18, 19, 20 and 15 and/or 16. OVKATE expressed CKs 6, 7, 8, 13, 17, 18, 19, 20 and 15 and/or 16. The expression of CK7, additional expression of vimentin, and clinical and histopathological findings enabled us to confirm that six cell lines had been established from primary ovarian cancers. Two of the six cell lines were positive for CK20, although CK20 was not expressed in the original tumours. The heterotransplanted tumours produced by CK20-positive cells also expressed CK20. This is the first report of ovarian carcinoma cell lines that express CK20 irrespective of their histological type. CK20 has been found in all colon carcinoma cell lines, but only in the mucinous type of ovarian tumours. These new ovarian carcinoma cell lines will therefore provide a relevant experimental system for elucidating the regulatory control mechanisms of intermediate filament expression. PMID- 9328140 TI - An immunohistochemical study of altered immunomodulatory molecule expression in head and neck squamous cell carcinoma. AB - For the presentation of peptide antigens to cytotoxic CD8+ T lymphocytes of the immune system, the expression of human leukocyte antigen (HLA) class I molecules on the cell surface is necessary. There is increasing evidence that surface HLA class I antigen expression is altered in a variety of human tumours by either loss or down-regulation of these molecules, which may be a strategy for evasion of immunosurveillance by malignant cells. This study has examined the expression of HLA class I molecules in head and neck squamous cell carcinoma (HNSCC) specimens by immunohistochemistry, using a wide panel of antibodies directed against allele-specific as well as monomorphic determinants of these molecules. The expression of TAP proteins, HLA-DR and the co-stimulatory molecule ICAM-1 were also studied. In addition, the expression of the tumour-associated antigens (TAA) p53 and MAGE genes was determined. Aberrant allelic expression of HLA class I antigens was detected in 17 out of 34 (50%) of the specimens stained, whereas HLA class I expression determined by W6/32 staining was found to be heterogeneous in only 2 out of 34 (6%) cases. Decreased expression of ICAM-1 was observed in 12 out of 34 (35%) tumour specimens and de novo expression of HLA-DR (HLA class II) by carcinoma cells in 13 out of 34 (38%) cases. Aberrant expression of HLA class I antigens was frequently observed in cases in which MAGE genes and p53 overexpression were detected. The altered expression of these immunomodulatory molecules in HNSCC may affect prognosis and has important implications for peptide-based immunotherapy strategies for these patients. PMID- 9328141 TI - Low nicotinamide mononucleotide adenylyltransferase activity in a tiazofurin resistant cell line: effects on NAD metabolism and DNA repair. AB - Poly(ADP-ribose) polymerase (PADPRP), which uses NAD to synthesize ADP-ribose polymers, is activated by DNA strand breaks and mediates cellular responses to DNA damage. The consequences of low cellular NAD levels in a cell line deficient in nicotinamide mononucleotide adenylyltransferase (NMNAT), an enzyme essential for NAD biosynthesis, were investigated by assessing NAD metabolism and DNA repair after treatment with alkylating agents. A tiazofurin-resistant L1210 cell line (TZR) was isolated. NAD levels were approximately 5933 and 3375 pmol mg(-1) protein for parental (wild type, WT) and TZR cells respectively, and NMNAT levels were reduced by > 95%. TZR cells were more sensitive to temozolomide (TM) and 1 methyl-3-nitro-1-nitroso-guanidine (MNNG), particularly at concentrations that caused > 50% NAD depletion. TM and MNNG treatment decreased NAD levels in both cell lines, but took longer to return to control levels in TZR cells. For example, MNNG (5 microM), depleted NAD levels at 6 h to approximately 4512 (WT) and 1442 (TZR) pmol mg(-1) protein; however, NAD levels had returned to control levels by 8 h in WT cells, but were not restored by 16 h in TZR cells. Both cell lines were equisensitive to the growth-inhibitory effects of NU1025 per se (IC50 370 microM). Co-exposure of the cell lines to TM (100 microM) with increasing concentrations of NU1025 led to a synergistic enhancement of cytotoxicity, with IC50 values for NU1025 decreasing to 17 +/- 4 microM (TZR) and 37 +/- 6 microM (WT). A similar enhanced sensitivity to NU1025 (approximately 2.7-fold) was obtained when TZR cells were co-exposed to MNNG + NU1025. TM-induced DNA strand breaks were increased by co-incubation with NU1025, and again the TZR cell line showed increased sensitivity to NU1025. There were no significant changes in NMNAT activity in response to MNNG treatment over 24 h, either in the presence or in the absence of NU1025. These data demonstrate that modest decreases in cellular NAD levels can sensitize cells to alkylating agents and PADPRP inhibitors. PMID- 9328142 TI - Ethnic variation in the prevalence of a common NAD(P)H quinone oxidoreductase polymorphism and its implications for anti-cancer chemotherapy. AB - The NAD(P)H quinone oxidoreductase (NQO1:EC 1.6.99.2) is an important biotransformation enzyme system that is also known to metabolize important novel chemotherapeutic compounds. The gene that codes for this enzyme has recently been found to be polymorphic in humans. Here, we describe the ethnic distribution of the polymorphism and note that this may have implications for anti-tumour drug development and use. PMID- 9328143 TI - Beta-carotene inhibits rat liver chromosomal aberrations and DNA chain break after a single injection of diethylnitrosamine. AB - Beta-carotene (BC) has recently been found to possess potent anti-tumour activity in chemically induced rat liver carcinogenesis. In the present study, attempts have been made to understand the basic cytogenetic and molecular mechanism of the anti-tumour effect of BC by monitoring its effect on rat liver chromosomal aberrations (CAs) and DNA chain breaks during the early preneoplastic stage of diethylnitrosamine (DEN)-induced hepatocarcinogenesis in male rats. DNA chain breaks, however, can be detected with great sensitivity by exposing crude cell lysates to alkaline solutions and monitoring the rate of strand unwinding so that one strand break per chromosome can easily be detected. Supplementary BC, in basal diet (120 mg kg[-1]), was given to rats 15 days before carcinogenic threat with DEN. Under these experimental conditions, BC was found to afford a unique protection against DEN-induced CAs 96 h after DEN injection. Long-term treatment with BC also triggered a protective effect on induction of CAs 15, 30 or 45 days after DEN treatment, which was maximal on structural aberrations followed by numerical and physiological types. BC treatment for 15 days before DEN injection was found to offer a significant (P < 0.001) protection in the generation of single-strand breaks compared with DEN control. Thus, BC ranks as a potential chemopreventive agent for the future so far as chemical rat liver carcinogenesis is concerned. PMID- 9328145 TI - Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis. AB - A cell line (MRc40) resistant to the model quinone compound, menadione, has been isolated from a parental Chinese hamster ovary cell line (CHO-K1). The known relationship between menadione toxicity and glutathione (GSH) depletion led us to investigate whether the mechanism of resistance of MRc40 was related to alteration in GSH homeostasis. Intracellular concentrations of GSH and cysteine (CySH) were twofold and 3.2-fold greater in MRc40 than in CHO-K1. Following exposure to menadione, GSH and CySH were depleted, but subsequent recovery of thiols was more rapid and of greater magnitude in MRc40 than in CHO-K1. Twelve hours after exposure to menadione, the concentrations of GSH and CySH were 9.7- and 4.2-fold greater in MRc40 than in CHO-K1. Using nuclear magnetic resonance (NMR) spectroscopy, we observed the in situ removal of menadione from cell suspensions of CHO-K1 and MRc40. However, only in CHO-K1 did we observe concomitant depletion of NMR-visible GSH. We conclude that the perturbation of GSH metabolism contributes to the resistant phenotype and is an important characteristic of menadione-resistant CHO cells. PMID- 9328144 TI - Resistance to the new anti-cancer phospholipid ilmofosine (BM 41 440). AB - The thioether phospholipid ilmofosine (BM 41 440) is a new anti-cancer drug presently undergoing phase II clinical trials. Because resistance to anti-tumour drugs is a major problem in cancer treatment, we investigated the resistance of different cell lines to this compound. Here we report that the multidrug resistant cell lines MCF7/ADR, CCRFNCR1000, CCRF/ADR500, CEM/VLB100 and HeLa cell lines transfected with a wild-type and mutated (gly/val185) multidrug resistance 1 gene (MDR1) are cross-resistant to ilmofosine compared with the sensitive parental cell lines. In CEMNM-1 cells, in which the resistance is associated with an altered topoisomerase II gene, no cross-resistance to ilmofosine was observed. Ilmofosine is not capable of modulating multidrug resistance and neither does it reduce the labelling of the P-glycoprotein (P-gp) by azidopine nor alter ATPase activity significantly. The resistance to ilmofosine in multidrug-resistant CCRF/VCR1000 cells cannot be reversed by the potent multidrug resistance modifier dexniguldipine-HCI (B8509-035). A tenfold excess of ilmofosine does not prevent the MDR-modulating effect of dexniguldipine-HCl. Treatment of cells with ilmofosine does not alter the levels of MDR1 mRNA. Long-term treatment of an ilmofosine-resistant Meth A subline with the drug does not induce multidrug resistance, indicating that ilmofosine does not increase the level of P-gp. Determination of the MDR2 mRNA levels in the cells revealed that the resistance pattern to ilmofosine is not correlated with the expression of this gene. It is concluded, therefore, that multidrug-resistant cells are cross-resistant to ilmofosine and that the compound is not a substrate of Pgp. No association between the expression of the MDR2-encoded P-gp and resistance to ilmofosine was observed. It is supposed that MDR1-associated alterations in membrane lipids cause resistance to ilmofosine. PMID- 9328146 TI - Influence of a haematoporphyrin derivative on the protoporphyrin IX synthesis and photodynamic effect after 5-aminolaevulinic acid sensitization in human colon carcinoma cells. AB - Haematoporphyrin derivatives (HPDs) are potent sensitizers in photodynamic therapy (PDT), associated with prolonged skin photosensitivity. 5-Aminolaevulinic acid (5-ALA), a natural precusor of haem, is converted intracellularly into the photosensitive agent protoporphyrin IX (PPIX), causing direct cytotoxicity after laser light irradiation but limited skin photosensitivity over 1-2 days and higher tumour selectivity. Unfortunately, the use of 5-ALA in PDT has been shown to cause only superficial tissue necrosis. Therefore, a combination of HPD and 5 ALA could be of great clinical value in the treatment of tumours if a synergistic effect of both sensitizers on tumour cell necrosis with less skin photosensitivity could be demonstrated. Human colon adenocarcinoma cells (HT-29) were cultured with either HPD or 5-ALA alone, simultaneously for 24 h with 5-ALA and HPD or in succession with 5-ALA (18 h) followed by HPD (6 h at different concentrations. Intracellular PPIX concentrations were determined by high performance thin-layer chromatography. Furthermore, PDT was performed with an incoherent light source (lambda = 580-740 nm) using a light dose of 30 J cm(-2) and an output power of 40 mW cm(-2). The intracellular PPIX concentration correlated well with 5-ALA drug dose and incubation time and was highest after single 5-ALA sensitization. In the presence of HPD, either simultaneously or sequentially, PPIX decreased significantly. The PDT effect after simultaneous incubation with both sensitizers for 24 h was not superior to incubation with HPD alone. If 5-ALA incubation (18 h) was followed by HPD (6 h) cytotoxicity after PDT was higher than with either single drug. 5-ALA (80 microg ml(-1)) led to a decrease in tumour cell viability by 40%. A similar effect could be observed when 5-ALA and HPD were sequentially combined allowing for a reduction of the 5-ALA dose from 80 microg ml(-1) in the absence of HPD to 60 microg ml(-1) and 5 microg ml(-1) together with 0.5 microg ml(-1) and 2 microg ml(-1) HPD respectively. We speculate that the enhanced PDT effect after the combined administration of 5-ALA and HPD to cultures of colon carcinoma cells should be even more impressive in the tumour in vivo, since HPD primarily targets the tumour microvasculature and secondarily tumour cells. PMID- 9328147 TI - Vitamin D analogues up-regulate p21 and p27 during growth inhibition of pancreatic cancer cell lines. AB - To obtain information regarding the growth-inhibitory effect of 1,25 dihydroxyvitamin D3 and its non-calcaemic analogue 22-oxa-1,25-dihydroxyvitamin D3 on pancreatic cancer cell lines, differences in the effects of G1-phase cell cycle-regulating factors were studied in vitamin D-responsive and non-responsive cell lines. Levels of expression of cyclins (D1, E and A), cyclin-dependent kinases (2 and 4) and cyclin-dependent kinase inhibitors (p21 and p27) were analysed by Western blotting after treatment with these compounds. In the responsive cells (BxPC-3, Hs 700T and SUP-1), our observations were: (1) marked up-regulation of p21 and p27 after 24 h treatment with 10(-7) mol l(-1) 1,25 dihydroxyvitamin D3 and 22-oxa-1,25-dihydroxyvitamin D3; and (2) marked down regulation of cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors after 7 days' treatment. In non-responsive cells (Hs 766T and Capan 1), no such changes were observed. In conclusion, vitamin D analogues up-regulate p21 and p27 as an early event, which in turn could block the G1/S transition and induce growth inhibition in responsive cells. PMID- 9328148 TI - Expression of the DNA mismatch repair proteins hMLH1 and hPMS2 in normal human tissues. AB - hMLH1 and hPMS2 are part of the DNA mismatch repair complex. Mutations in these genes have been linked to hereditary non-polyposis colon cancer; they also occur in a variety of sporadic cancers. Western blot analysis and immunohistochemistry demonstrated that hMLH1 and hPMS2 are widely expressed nuclear proteins with a distribution pattern very similar to that previously described for hMSH2. These observations showing similar localization of hMLH1 and hPMS2 with hMSH2 are consistent with the biochemical function of these proteins in DNA mismatch repair. PMID- 9328149 TI - An experimental and mathematical model for the extravascular transport of a DNA intercalator in tumours. AB - A new in vitro model has been developed for investigating extravascular diffusion of therapeutic agents in tumour tissue. V79-171b or EMT6/Ak cells are grown on porous Teflon support membranes and submerged in a large reservoir of medium, to give diffusion-limited 'multicellular membranes' (MMs) c. 200 microm in thickness. MMs are histologically similar to multicellular spheroids, but their planar rather than spherical geometry facilitates direct measurement of the flux of radiolabelled agents through the multicellular structure. For [14C]urea, flux kinetics through V79-171b MMs was modelled as simple diffusion, yielding a diffusion coefficient in the MM (DMM) of 1.45 x 10(-6) cm2 s(-1), 11-fold lower than in culture medium. Flux of the 3H-labelled DNA intercalator 9-[3-(N,N dimethylamino)propylamino]acridine (DAPA) was dramatically slower than urea. Modelling this over the first 5 h gave a DMM of 1.3 x 10(-8) cm2 s(-1), but over longer times the kinetics was not consistent with simple diffusion. Flux of DAPA was markedly increased in the presence of 50 mM ammonium chloride, indicating that sequestration in acidic endosomes is a major impediment to flux. Accumulation in cytoplasmic vesicles was confirmed by fluorescence microscopy. The DAPA flux kinetics, with and without ammonium chloride, was well fitted by a reaction-diffusion model with reversible cellular uptake (modelled as binding), using uptake parameters determined in separate experiments with V79-171b single cell suspensions. This study demonstrates the utility of the MM model for determining extravascular transport parameters, and indicates that much of the impediment to diffusion of basic DNA intercalators in tumour tissue may arise from lysosomal sequestration rather than DNA binding. PMID- 9328150 TI - Tumour cells of extramammary Paget's disease do not show either p53 mutation or allelic loss at several selected loci implicated in other cancers. AB - Extramammary Paget's disease is a particular form of skin cancer of unknown histogenesis. To look for the genetic defects underlying the pathogenesis of this tumour, we have examined loss of heterozygosity (LOH), p53 and human papillomavirus (HPV) status, and the expression of c-erbB-2 and bcl-2 proteins in 14 cases. Unexpectedly, no LOH was detected at several loci commonly lost in other human cancers (namely 3p, 9p, 9q, 13q, 16q, 17p, and 17q) in 12 tumours examined. Altered p53 protein expression was entirely or mostly negative in all 14 cases. Direct sequencing of exons 5-8 of the p53 gene in eight cases revealed no mutation. Polymerase chain reaction amplification of the L1 gene of human papillomavirus (HPV) did not detect the virus that could inactivate p53 and retinoblastoma tumour-suppressor gene products. As expected, c-erbB-2 proto oncogene protein was overexpressed in six cases. The expression of bcl-2 was negative in all cases. The results presented in this study suggest that molecular events underlying extramammary Paget's disease differ from those of other common epithelial malignancies and that tumour-suppressor genes located in chromosome regions not examined in this study may be important. PMID- 9328151 TI - Expression of an antigen homologous to the human CO17-1A/GA733 colon cancer antigen in animal tissues. AB - The CO17-1A/GA733 antigen is associated with human carcinomas and some normal epithelial tissues. This antigen has shown promise as a target in approaches to passive and active immunotherapy of colorectal cancer. The relevance of animal models for studies of immunotherapy targeting this antigen in patients is dependent on the expression of the antigen on normal animal tissues. Immunohistoperoxidase staining with polyclonal rabbit antibodies to the human antigen revealed the human homologue on normal small intestine, colon and liver of mice, rats and non-human primates, whereas mouse monoclonal antibodies to the CO17-1A or GA733 epitopes on the human antigen did not detect the antigen. Polyclonal rabbit antibodies, elicited by the murine antigen homologue derived from recombinant baculovirus-infected insect cells, immunoprecipitated the antigen from mouse small intestine, colon, stomach, kidney and lung. The isolated recombinant murine protein bound polyclonal, but not monoclonal, antibodies to the human CO17-1A/GA733 antigen, and recombinant human antigen bound polyclonal antibodies elicited by the murine antigen homologue. Thus, the antigen homologue expressed by animal tissues is similar, but not identical, to the human antigen. These results have important implications for experimental active and passive immunotherapy targeting the CO17-1A/GA733 antigen. PMID- 9328152 TI - Predictive value of c-erbB-2, p53, cathepsin-D and histology of the primary tumour in metastatic breast cancer. AB - The value of various prognostic factors in breast cancer patients has been determined in a number of studies. Few reports have been published on the dependence of treatment outcome on histological and immunohistochemical characteristics in the primary tumour in patients with metastatic disease. We studied the incidence and prognostic value of histological and molecular abnormalities in the primary tumour of patients who had developed metastatic breast cancer. Eligible patients received a fluorouracil, epirubicin and cyclophosphamide (FEC) regimen either once a week or once every 4 weeks. Adequate specimens for various analyses were available from 127 patients. Median follow-up time of the patients ranged from 15 to 101 months. In this study, the histological grade of the malignancy best predicted response to chemotherapy (P < 0.0005). Most of the responses were observed in patients with grade 1 tumours; in this group, time to progression was delayed. C-erb B-2 gene amplification and oncoprotein expression had no predictive value. Neither p53 nor cathepsin-D predicted treatment outcome after chemotherapy. None of the factors had an effect on overall survival. Among breast cancer patients who received anthracycline containing chemotherapy, response to treatment correlated with histological grade. In patients with histological grade 1 breast cancer, the time to progression was longest. However, overall survival was not affected by histological grade nor the other parameters tested. In addition to histological grade, other prognostic factors that are not included in this study need to be identified to determine which patients with metastatic breast cancer would benefit from cytotoxic treatment. PMID- 9328153 TI - DT-diaphorase and cytochrome B5 reductase in human lung and breast tumours. AB - The level of expression of enzymes that can activate or detoxify bioreductive agents within tumours has emerged as an important feature in the development of these anti-tumour compounds. The levels of two such reductase enzymes have been determined in 19 human non-small-cell lung tumours and 20 human breast tumours, together with the corresponding normal tissue. DT-diaphorase (DTD) enzyme levels (both expression and activity) were determined in these samples. Cytochrome b5 reductase (Cytb5R) activity was also assessed. With the exception of six patients, the levels of DTD activity were below 45 nmol min(-1) mg(-1) in the normal tissues assayed. DTD tumour activity was extremely variable, distinguishing two different groups of patients, one with DTD activity above 79 nmol min(-1) mg(-1) and the other with levels that were in the same range as found for the normal tissues. In 53% of the lung tumour samples, DTD activity was increased with respect to the normal tissue by a factor of 2.4-90.3 (range 79-965 nmol min[-1] mg[-1]). In 70% of the breast tumour samples, DTD activity was over 80 nmol min(-1) mg(-1) (range 83-267 nmol min[-1] mg[-1]). DTD expression measured by Western blot correlated well with the enzyme activity measured in both tumour and normal tissues. The levels of the other reductase enzyme, Cytb5R, were not as variable as those for DTD, being in the same range in both tumour and normal tissue or slightly higher in the normal tissues. The heterogeneous nature of DTD activity and expression reinforces the need to measure enzyme levels in individual patients before therapy with DTD-activated bioreductive drugs. PMID- 9328154 TI - Enhanced expression of vascular endothelial growth factor in metastatic melanoma. AB - Tumour growth is dependent on angiogenesis. Vascular endothelial growth factor (VEGF) is a secreted endothelial cell-specific cytokine. VEGF is angiogenic in vivo and it also acts as a vascular permeability factor. VEGF is overexpressed in many skin disorders characterized by angiogenesis and increased vascular permeability. We investigated VEGF expression in 22 primary cutaneous melanomas, 33 melanoma metastases and six naevocellular naevi using immunohistochemistry. VEGF accumulated on the vascular endothelia in the normal dermis, suggesting that a constitutive low level of VEGF expression may regulate skin vessel function under normal physiological conditions. No VEGF was detected in the cells of naevocellular naevi or normal dermis. In contrast, 32% of the primary and 91% of the metastatic melanomas contained melanoma cells staining for VEGF. Expression of VEGF was more frequent in metastases than in primary melanomas (P <0.0001). Tumour-infiltrating inflammatory cells expressed VEGF in all melanomas. A high number of VEGF-expressing inflammatory cells was associated with high VEGF expression in melanoma cells (P = 0.003). Our results suggest that VEGF is up regulated during the course of melanoma progression and dissemination and that tumour-infiltrating cells expressing VEGF may contribute to the progression of melanoma. PMID- 9328155 TI - Bcl-2/Bax ratios in chronic lymphocytic leukaemia and their correlation with in vitro apoptosis and clinical resistance. AB - The bcl-2 gene is overexpressed in the absence of gene rearrangements in most cases of B-cell chronic lymphocytic leukaemia (B-CLL) and the proto-oncogene product Bcl-2 has been shown to be a regulator of apoptosis. The activity of this protein is opposed by Bax, a homologous protein that accelerates the rate of cell death. B-lymphocyte Bcl-2 and Bax protein levels were found to be significantly altered in B-CLL and increased Bcl-2/Bax ratios were observed in both the treated and untreated patients compared with those of normal controls. These alterations were particularly pronounced in those treated patients found to be clinically unresponsive to chemotherapy. In order to determine whether Bcl-2/Bax ratios affected cell survival via an anti-apoptotic mechanism, cell death was induced in B-CLL cells in vitro using chlorambucil, and apoptosis was monitored by Annexin V and propidium iodide staining. Confirmation that the labelled cells were apoptotic was achieved by morphological assessment of cytospin preparations of cell-sorted populations. Drug-induced apoptosis in B-CLL cells was inversely related to Bcl-2/Bax ratios. PMID- 9328156 TI - Concomitant i.v. and oral clodronate in the relief of bone pain--a double-blind placebo-controlled study in patients with prostate cancer. AB - Fifty-seven patients with advanced prostate cancer resistant to first-line hormonal therapy were treated with estramustine and additionally randomized for treatment with clodronate or placebo. Clodronate treatment was started with 5 days intravenous administration (300 mg day[-1]) and followed by oral treatment (1.6 g day[-1]) for 12 months. Skeletal pain relief was only about 10% better in the clodronate than in the placebo group. The results do not support the superiority of combined intravenous and oral treatment with clodronate compared with oral administration only. PMID- 9328157 TI - Cardiotoxicity from intensive chemotherapy combined with radiotherapy in breast cancer. AB - Cardiac function was evaluated in 86 breast cancer patients after standard chemotherapy, followed by ablative chemotherapy and chest irradiation. One patient died of subacute heart failure 3 months after ablative chemotherapy. At a minimum of 1 year's follow-up (range 1-11 years) left vertricular ejection fraction (LVEF) was marginally abnormal in 4 of 27 disease-free survivors. One exceptional patient who received two transplantations is alive, with serious heart failure occurring after the second ablative chemotherapy. Including this patient, the percentage of patients free of clinical and subclinical cardiac dysfunction at 7 years is 78% (95% CI 61-95%). After ablative chemotherapy, cardiotoxicity was rarely life-threatening. The impact of subclinical cardiotoxicity in the long term is not clear and needs continued evaluation. PMID- 9328158 TI - The bioavailability of oral GI147211 (GG211), a new topoisomerase I inhibitor. AB - Topoisomerase I inhibitors are new compounds of interest for cancer chemotherapy. We performed a study with GI147211, a new semisynthetic camptothecin analogue, to determine the absolute bioavailability of the drug given orally. Patients with a histologically confirmed diagnosis of a solid tumour refractory to standard forms of therapy were eligible for the study. GI147211 was given orally on day 1 and as a 30-min infusion daily on days 2-5. The treatment course was repeated every 3 weeks. In subsequent patient cohorts, the dose of the oral formulation was escalated from 1.5 mg m(-2) to 6.0 mg m(-2); the dose for i.v. administration was fixed at 1.2 mg m(-2). Plasma pharmacokinetics was performed on day 1 and 2 of the first course and on day 1 of the second course using a validated high performance liquid chromatographic assay. Nineteen patients were entered into the study; one patient was not evaluable because the treatment course was stopped prematurely. Eighteen patients received a total of 47 treatment courses. The absolute bioavailability of GI147211 averaged 1.3 +/- 5.2%. Drug appeared quickly in plasma with a median Tmax at 0.5 h. Fasting or fed state had no significant influence on the bioavailability of GI147211. The terminal half-life after administration of oral GI147211 was 6.85 +/- 3.13 h, similar to the half-life after intravenous administration. The major toxicities were neutropenia and thrombocytopenia. Nadirs for neutropenia and thrombocytopenia occurred on day 8 and day 15 respectively. Other toxicities predominantly consisted of mild and infrequent nausea and vomiting, and fatigue. The oral administration of the drug is well tolerated. Oral administration of topoisomerase I inhibitor GI147211 results in a low bioavailability with relatively wide interpatient variation. The intravenous route of administration is advised for further development of this promising topoisomerase I inhibitor. PMID- 9328160 TI - Colon cancer in the elderly: evidence for major improvements in health care and survival. AB - Time trends in therapeutic approaches and in the prognosis of colon cancer for patients aged 75 years and above have been investigated in comparison with corresponding trends for younger patients using a population-based series of 2089 colon cancer patients diagnosed between 1976 and 1990 in the Cote-d'Or area (478,000 inhabitants), Burgundy, France. Significant progress has been achieved in the management of patients with colon cancer in both age groups, but trends have been more noticeable in patients aged 75 years and above. In the elderly, the proportion of cancers limited to the digestive tract wall showed a 3-year average increase of 2.8% (P = 0.02) and the frequency of curative surgery an average increase of 8.6% (P < 0.001), so that it was performed in 80% of cases in the last 3-year period. Operative mortality decreased by 2.5% between 3-year periods (P < 0.004). Crude 5-year survival rates in elderly patients increased from 15% in the 1976-78 period to 29% in the 1985-87 period (P < 0.001), the corresponding figures being 36% and 44% (P > 0.10) in younger patients. PMID- 9328159 TI - Topoisomerase I inhibitors: the relevance of prolonged exposure for present clinical development. AB - Topoisomerase I inhibitors constitute a new class of anti-cancer agents. Recently, topotecan and irinotecan were registered for clinical use in ovarian cancer and colorectal cancer respectively. Cytotoxicity of topoisomerase I inhibitors is S-phase specific, and in vitro and in vivo studies have suggested that, for efficacy, prolonged exposure might be more important than short-term exposure to high concentration. Clinical development of those topoisomerase I inhibitors that have reached this stage is also focused on schedules aiming to achieve prolonged exposure. In this review, we summarize all published preclinical studies on this topic for topoisomerase I inhibitors in clinical development, namely 20-S-camptothecin, 9-nitro-camptothecin, 9-amino camptothecin, topotecan, irinotecan and GI147211. In addition, preliminary data on clinical studies concerning this topic are also reviewed. The data suggest that prolonged exposure may indeed be relevant for anti-tumour activity. However, the optimal schedule is yet to be determined. Finally, clinical data are yet too immature to draw definitive conclusions. PMID- 9328162 TI - Induction of chromosome-specific aneuploidy and micronuclei in human lymphocytes by metabolites of 1,3-butadiene. AB - 1,3-Butadiene is a carcinogen in rodents, but its potential carcinogenicity to humans remains controversial. Numerous studies have shown that butadiene and its metabolites cause sister chromatid exchanges in vitro and in vivo. To test for other types of genotoxicity, the micronucleus assay and fluorescence in situ hybridization (FISH) have been used to detect chromosome damage in human lymphocytes caused by two reactive metabolites of butadiene, diepoxybutane (DEB) and monoepoxybutene (MEB). DEB (0.5-5.0 microM) significantly increased micronucleus formation 4- to 6-fold (P <0.01) and MEB (1-500 microM) by 2- to 4 fold (P <0.01) over control levels. The ability of DEB and MEB to induce aneuploidy of chromosomes 7, 8, 12, and X was examined using dual-color FISH in both interphase and metaphase cells. These chromosomes were chosen because of their involvement in leukemogenesis. Both DEB and MEB caused dose-dependent increases in hyperdiploidy of chromosomes 12 and X, but had no discernible effect on chromosomes 7 and 8. These results suggest that DEB and MEB cause chromosome specific aneuploidy in human cells. If formed in sufficient amounts, DEB and MEB may produce chromosome damage of the type found in leukemia following exposure to butadiene. PMID- 9328163 TI - Identification of benzene oxide as a product of benzene metabolism by mouse, rat, and human liver microsomes. AB - Benzene is a ubiquitous environmental pollutant that is known to cause hematotoxicity and leukemia in humans. The initial oxidative metabolite of benzene has long been suspected to be benzene oxide (3,5-cyclohexadiene-1,2 oxide). During in vitro experiments designed to characterize the oxidative metabolism of [14C]benzene, a metabolite was detected by HPLC-radioactivity analysis that did not elute with other known oxidative metabolites. The purpose of our investigation was to prove the hypothesis that this metabolite was benzene oxide. Benzene (1 mM) was incubated with liver microsomes from human donors, male B6C3F1 mice, or male Fischer-344 rats, NADH (1 mM), and NADPH (1 mM) in 0.1 M sodium phosphate buffer (pH 7.4) and then extracted with methylene chloride. Gas chromatography-mass spectrometry analysis of incubation extracts for mice, rats, and humans detected a metabolite whose elution time and mass spectrum matched that of synthetic benzene oxide. The elution time of the benzene oxide peak was approximately 4.1 min, while phenol eluted at approximately 8 min. Benzene oxide also coeluted with the HPLC peak of the previously unidentified metabolite. Based on the 14C activity of this peak, the concentration of benzene oxide was determined to be approximately 18 microM, or 7% of total benzene metabolites, after 18 min of incubation of mouse microsomes with 1 mM benzene. The metabolite was not observed in incubations using heat-inactivated microsomes. This is the first demonstration that benzene oxide is a product of hepatic benzene metabolism in vitro. The level of benzene oxide detected suggests that benzene oxide is sufficiently stable to reach significant levels in the blood of mice, rats, and humans and may be translocated to the bone marrow. Therefore benzene oxide should not be excluded as a possible metabolite involved in benzene-induced leukemogenesis. PMID- 9328161 TI - Effect of hepatitis C and B virus infection on risk of hepatocellular carcinoma: a prospective study. AB - To assess whether there is an additive effect between chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection on the development of hepatocellular carcinoma (HCC), 400 consecutive cirrhotic patients were followed prospectively with periodic abdominal ultrasound examination and measurement of serum alpha fetoprotein (AFP) level every 4 months. During a follow-up of 1185 person-years, 80 (20%) patients developed HCC, with an annual incidence of 6.8%. The annual incidence was 2.0% in patients negative for hepatitis B surface antigen (HBsAg) and antibodies to HCV (anti-HCV), 6.6% in patients with HBsAg alone, 7.0% in patients with anti-HCV alone and 13.3% in patients co-infected with HBV and HCV. There was a positive linear trend in the annual incidence of HCC among patients without either marker, patients with single viral infection and patients with dual viral infection (P[for trend] < 0.0001). Cox's proportional hazard model indicated that HCV/HBV co-infection [hazard ratio (HR), 6.41; 95% confidence interval (CI), 1.80-22.80], anti-HCV alone (HR, 3.74; 95% CI, 1.07-13.07) and HBsAg alone (HR, 4.06; 95% CI, 1.23-13.34) were independently risk factors of HCC. In conclusion, there is an additive and independent effect modification of HCV and HBV infection on HCC development. PMID- 9328165 TI - Uracil misincorporation in human DNA detected using single cell gel electrophoresis. AB - Poor folate status may be important in the aetiology of several epithelial cell malignancies including cancer of the uterine cervix. Folic acid is essential in the synthesis of purine nucleotides and the pyrimidine nucleoside thymidine and it is probable that imbalances in these DNA precursors negatively effect DNA stability and may ultimately lead to malignant transformation. The development of a modified 'comet assay' using the bacterial DNA repair enzyme uracil DNA glycosylase, to detect misincorporated uracil in human DNA is reported here. The effect of perturbing folic acid and deoxyuridine levels on uracil misincorporation in normal human lymphocytes and cultured human tumour cells was investigated using this assay. HeLa cells and peripheral human lymphocytes incubated as agarose-embedded nucleoids, with 1 unit of uracil DNA glycosylase per microg of DNA, contained low levels of uracil in their DNA. Both HeLa cells and stimulated human lymphocytes cultured in folate-deficient medium were growth arrested. Incubating human lymphocytes in folate-deficient medium significantly increased the level of uracil detected compared with control cells. HeLa cells showed an increase in non-specific DNA damage (strand breaks). Deoxyuridine (100 microM) significantly increased the level of uracil detected in the DNA of both folate-deficient and control HeLa cells. It appears that this modified comet assay specifically detects misincorporated uracil in single human cells. It should, therefore, prove valuable in determining the role of folic acid status in DNA instability and cancer. PMID- 9328164 TI - The influence of cellular apoptotic capacity on N-nitrosodimethylamine-induced loss of heterozygosity mutations in human cells. AB - The induction of loss of heterozygosity (LOH) by the environmental carcinogen N nitrosodimethylamine (NDMA), and the factors that influence the recovery of LOH mutations were studied in two directly related human lymphoblastoid cell lines, AHH-1 (h2E1.v2) and MCL-5. Initially, the NDMA-induced mutation frequency at the heterozygous tk locus in AHH-1 cells was observed to be 5-fold higher in AHH-1 compared with MCL-5. Molecular analysis of NDMA-induced TK- mutants indicated that the induced mutant fraction attributable to small intragenic mutations was similar in both cell lines. However, the induced mutant fraction, because of LOH, was 18-fold greater in AHH-1. In addition, LOH mutations were more extensive among TK- mutants derived from AHH-1 cells. We hypothesized that the increased recovery of large LOH mutations in AHH-1 cells could be attributable to reduced apoptotic capacity, as it has been reported that AHH-1 cells carry a heterozygous mutation in the p53 locus, whereas MCL-5 cells are homozygous wild-type. Analysis of the kinetics of apoptosis showed that the apoptotic response of the AHH-1 cell line was diminished and delayed compared with MCL-5. Based on the analyses presented here, and several recent reports, it is suggested that the recovery of LOH mutations in p53 deficient cell lines is affected not only by abnormalities in cellular apoptotic response, but also involves a number of p53-mediated responses to DNA damage. PMID- 9328166 TI - Effects of long term dietary phenethyl isothiocyanate on the microsomal metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol in F344 rats. AB - Phenethyl isothiocyanate (PEITC), a cruciferous vegetable component, inhibits lung tumor induction by the tobacco specific nitrosamine, 4-(methylnitrosamino)-1 (3-pyridyl)-1-butanone (NNK). To gain insight into the mechanism of PEITC lung tumor inhibition, we examined, in male F344 rats, the effects of dietary PEITC (3 micromol/g NIH-07 diet) in combination with NNK treatment (1.76 mg/kg, s.c., three times a week) for 4, 12 and 20 weeks on liver and lung microsomal metabolism of NNK and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of NNK and also a lung carcinogen. This was compared with rats fed NIH-07 diet, without PEITC, and treated with NNK alone or saline. The protocol was identical to that employed for inhibition of lung tumorigenesis by PEITC. We observed decreased rates of alpha-hydroxylation of NNK and NNAL in lung microsomes of 4-, 12- and 20-week PEITC + NNK treated rats compared with those treated with NNK or saline. NNK treatment alone also decreased lung alpha methylene hydroxylation of NNK. Long-term NNK + PEITC administration did not significantly affect liver oxidative metabolism of NNK or NNAL, and did not affect the rate of glucuronidation of NNAL in liver microsomes when compared with rats treated with NNK or saline. Thus, PEITC selectively inhibited lung metabolic activation of NNK and NNAL. These results support the hypothesis that PEITC inhibits NNK-induced lung tumors by inhibiting metabolic activation of NNK in the lung. This study also demonstrated that PEITC inhibits lung alpha-hydroxylation of NNAL; this may play a role in PEITC inhibition of lung tumorigenesis by NNK. PMID- 9328167 TI - Anti-carcinogenic activity of simvastatin during the promotion phase of radiation induced mammary tumorigenesis of rats. AB - Pregnant Wistar-MS rats received whole body irradiation with 2.6 Gy gamma-rays from a 60Co source at day 20 of pregnancy. Control rats were fed a basal diet and were implanted with a diethylstilbestrol (DES) pellet at 30 days after weaning. In the experimental group, rats were fed a diet containing simvastatin immediately after weaning and received a DES pellet at 30 days after weaning. A high incidence of total mammary tumours was observed in the rats fed the control diet and treated with DES for 1 year. The administration of dietary simvastatin together with DES treatment significantly decreased the incidence of mammary tumours. The development of adenocarcinoma in the control rats was significantly higher than that in the rats fed the simvastatin diet. After the administration of simvastatin to the experimental group for 1 year, the serum estradiol-17beta concentration in these rats was markedly reduced, but that of prolactin was not. No significant difference was seen in the development of the mammary glands between rats fed the control diet and those fed the simvastatin diet by whole mount observations. Simvastatin feeding produced an increased development of ER- PgR- tumours and a reduced incidence of ER+ PgR+ tumours. These findings suggest that simvastatin has a potent preventive activity during the DES-dependent promotion/progression phase of radiation-induced mammary tumorigenesis. PMID- 9328168 TI - Mechanism of action of dietary chemoprotective agents in rat liver: induction of phase I and II drug metabolizing enzymes and aflatoxin B1 metabolism. AB - A range of potential chemoprotective agents, most of them natural dietary constituents, has been examined for ability to modulate both phase I (cytochrome P450 1A1, 1A2, 2B1/2, 2C11, 2E1, 3A, 4A) and phase II drug metabolizing enzymes (glutathione S-transferases, in particular subunits Yc2 and P, aflatoxin B1 aldehyde reductase and quinone reductase) in rat liver. In addition to assays of total enzyme activity and Western blots for individual isozymes, the ability of microsomes to metabolize aflatoxin B1, and of cytosols to conjugate aflatoxin B1 (AFB1)-epoxide to GSH and to produce AFB1-dialcohol, were measured. Induction of gamma-glutamyl transpeptidase activity was examined by histochemistry. Differing patterns of induction were observed, reflecting differences in the control of expression of the individual enzymes studied. Of the compounds examined, butylated hydroxytoluene, ethoxyquin, indole-3-carbinol and phenethyl isothiocyanate were the most potent bifunctional agents (inducing both phase I and II activities). Oltipraz, while only weakly inducing CYP1A2 and 2B1/2, was a potent inducer of phase II enzymes. Caffeic acid, garlic oil, sinigrin and propyl gallate all showed some ability to induce phase II enzymes. 4-Methyl catechol, alpha-tocopherol and red wine decreased certain phase I enzyme activities, while inducing total GST activity. Butylated hydroxytoluene, ethoxyquin, garlic oil and indole-3-carbinol induced gamma glutamyltranspeptidase in periportal hepatocytes. Particularly because of their ability to induce the detoxifying activities of glutathione S-transferase Yc2 and aldehyde reductase, butylated hydroxytoluene, ethoxyquin, indole-3-carbinol, oltipraz, phenethyl isothiocyanate and sinigrin will be effective blocking agents in rodents, if administered prior to AFB1. While these studies indicate the relative contributions of phase I and II metabolism in the overall protective effect in rat, care should be taken that a similar balance is achieved in man, and that relevant enzymes or iso forms are induced. PMID- 9328169 TI - High acetaldehyde levels in saliva after ethanol consumption: methodological aspects and pathogenetic implications. AB - Chronic ethanol ingestion leads to an enhanced risk of upper gastrointestinal tract cancer. Although many hypotheses for the tumor promoting effect of alcohol exist, the pathogenetic mechanisms remain unclear since alcohol in itself is not carcinogenic. Acetaldehyde, the first metabolite of ethanol, has been shown to have multiple mutagenic effects and to be carcinogenic to animals. Previous research has revealed that acetaldehyde can be formed from ethanol via microbial alcohol dehydrogenase. Thus, at least part of the proposed tumorigenic effect of ethanol may be linked to local production of acetaldehyde from ethanol by oral microflora. In this study we demonstrate the production of marked amounts of acetaldehyde in saliva after ingestion of moderate amounts of ethanol. Considerable inter individual variation in acetaldehyde production capacity is also shown. In vivo acetaldehyde production is significantly reduced after a 3 day use of an antiseptic mouthwash (chlorhexidine). In vitro acetaldehyde production was shown to be linear in time, inhibited by 4-methylpyrazole and it could not be saturated under ethanol conditions that are relevant in vivo. There was a significant positive correlation between salivary acetaldehyde production in vitro and in vivo. We conclude, that the microbial formation of acetaldehyde in saliva could be one explanation for the tumor promoting effect of ethanol on the upper gastrointestinal tract. Moreover, this may support the epidemiological finding, that poor oral hygiene is an independent risk factor for oral cavity cancer. PMID- 9328170 TI - Resistance of Copenhagen rats to chemical induction of glutathione S-transferase 7-7-positive liver foci. AB - Copenhagen (Cop) rats are completely resistant to the chemical induction of mammary adenocarcinomas, but their susceptibility to hepatocarcinogenesis is virtually unknown. Rat liver is a well-characterized and easily manipulated tissue in which to study carcinogenesis. Therefore, if Cop rats are resistant to hepatocarcinogenesis, studies into resistance mechanisms may be feasible. Male Cop and F344 rats, 7-8 weeks old, were initiated using either N nitrosodiethylamine (DEN) (200 mg/kg, i.p.) or a two-thirds partial hepatectomy (PH) followed by N-methyl-N-nitrosourea (MNU) (60 mg/kg, i.p.). The rats were then promoted using a modified resistant hepatocyte (RH) protocol (a combination of four doses of 2-acetylaminofluorene (2-AAF) and a single dose of CCl4 that provides a selective mitotic stimulus for initiated cells). Six weeks after initiation the rats were killed and liver sections were stained for glutathione S transferase 7-7 (GST 7-7), a marker for putative preneoplastic hepatocytes. Cop rats were found to be highly resistant, having a approximately 9- and approximately 27-fold smaller percentage of liver area occupied by GST 7-7 positive foci than susceptible F344 rats following initiation by DEN and MNU respectively. Furthermore, gross liver nodules did not form in any of the Cop rats, whereas all F344 rat livers contained nodules. Hepatic necrosis caused by DEN during initiation, and CCl4 during promotion is necessary to stimulate compensatory hepatocyte division. We demonstrated that these agents do indeed increase serum transaminase levels and produce histologic evidence of necrosis in Cop rats. In order for liver foci to grow rapidly in the RH protocol, the surrounding normal hepatocytes must be mito-inhibited by 2-AAF. We found that the degree of mito-inhibition of normal hepatocytes by 2-AAF is the same in Cop and F344 rats. These results show that the Cop rat is highly resistant to the chemical induction of putative preneoplastic liver foci and nodules. PMID- 9328171 TI - Smad4 (homolog of human DPC4) and Smad2 (homolog of human JV18-1): candidates for murine lung tumor resistance and suppressor genes. AB - In this study we investigated the mouse mad-related genes, Smad4/Dpc4 and Smad2 (homolog of JV18-1), as candidates for involvement in lung tumor resistance and suppression. These genes are located in a region of mouse chromosome 18 that is syntenic with human 18q21.1, where several genes that are mutated in various cancers have been mapped. A newly identified murine lung tumor resistance locus, Par2 has also been mapped to this region of chromosome 18. We found no mutations in the coding regions of either gene in 11 lung tumors from B6CF1 (C57BL/6 x BALB/c) mice by RT-PCR and SSCP/RFLP, suggesting that these genes are not mutated in lung carcinogenesis in this strain. Moreover, loss of heterozygosity in this region of chromosome 18 was not detected in 28 lung adenocarcinomas from B6CF1 mice, 17 lung adenocarcinomas from B6C3F1 mice or 18 lung adenocarcinomas from AB6F1 mice. These data provide evidence that a 'classical' tumor suppressor gene for mouse lung carcinogenesis in these strains does not reside in this region. In order to investigate Smad4/Dpc4 and Smad2 as candidates for the Par2 resistance locus mapped to this region, we also sequenced the coding regions of both genes in cDNA from normal lungs of A/J, BALB/c and C57BL/6 inbred strains of mice. No polymorphisms were detected in the coding region of Smad4. In Smad2, two sequence polymorphisms were identified that are not in the conserved regions of the gene. Northern blot analysis revealed no differential expression in normal lung tissue among the three strains for either gene. Thus, in this study we found no evidence that either Smad4 or Smad2 represents the Par2 lung tumor resistance locus or is a lung tumor suppressor gene in the B6CF1 mice. PMID- 9328172 TI - Promotion of dimethylbenz[a]anthracene-initiated mammary carcinogenesis by iron in female Sprague-Dawley rats. AB - Iron body-stores and iron dietary intake have been sporadically reported to increase the risk of cancer in humans. To investigate the effect of iron on the development of mammary tumors, female Sprague-Dawley rats were given dimethylbenz[a]anthracene (DMBA) (5 mg/kg, i.g., 1x) at 55 days of age. Eight days later, rats received iron(II) sulfate s.c. (50 micromol/kg, 2x/week) for 53 weeks. Mammary tumors started to appear 6-8 weeks after DMBA initiation. At 20 weeks after DMBA treatment, iron(II) increased mammary tumor frequency twofold (11/30 versus 5/30 with DMBA alone). Tumor frequency increased with time and was significantly higher in iron-promoted rats after 40 weeks of treatment (24/30 versus 11/30, P = 0.001). Also, mammary tumors in iron-promoted rats were significantly larger than in DMBA-only rats at 20 weeks after initiation (P = 0.04) and this difference remained significant through the observation time point at 40 weeks. Iron could be detected histochemically in the stromal connective tissue, but not in the epithelial cells of mammary carcinomas. Mammary tumors in the DMBA-only group were mostly adenomas and adenocarcinomas, while those promoted by iron sulfate included fibroadenomas, adenomas and adenocarcinomas. Thus, iron(II) administered s.c. subsequent to DMBA initiation, greatly accelerated mammary carcinogenesis, implying its promoting activity for mammary tissue of female rats. PMID- 9328173 TI - Cigarette smoking induces an increase in oxidative DNA damage, 8 hydroxydeoxyguanosine, in a central site of the human lung. AB - To evaluate the effect of cigarette smoking on oxidative stress in lung tissues, we compared the levels of a type of oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OH-dG), in tissue obtained from the central sites of lungs from 14 current smokers, seven ex-smokers and nine non-smokers. The mean level of 8-OH-dG in the lung tissues from smokers was 1.43-fold higher than that of the non-smokers (the difference was statistically significant, P = 0.0262). A positive correlation between the 8-OH-dG levels in normal lung tissues and the Brinkman index was obtained from smokers and ex-smokers (r = 0.525; P = 0.0134). A positive correlation was also obtained for the 8-OH-dG levels in normal lung tissues and the number of cigarettes smoked per day (r = 0.436; P = 0.0132). These results suggest that oxidative DNA damage is induced in lung DNA by cigarette smoking. PMID- 9328174 TI - Improved 32P-postlabelling assay for the quantification of the major platinum-DNA adducts. AB - For the improvement of chemotherapy with platinum (Pt)-containing drugs a sensitive assay to detect the induced Pt-DNA adducts is needed. Therefore, the 32P-postlabelling assay, described by Blommaert and Saris (Nucleic Acids Res., 1995, 23, 1300-1306), to detect the major adducts Pt-GG and Pt-AG has substantially been improved and compared with ELISA and AAS. For the quantification of the adducts, TpT was added as an internal standard immediately after isolation of the Pt-adducts from digested DNA samples. It was found that 32P-labelling of both GpG and ApG, the dinucleotides obtained after deplatination of the adducts, was equally efficient as that of TpT. To isolate the Pt-adducts on basis of a positive charge, the pH of DNA digests was adjusted to approximately 3 prior to separation by strong cation-exchange chromatography. For the subsequent deplatination a volume of only 12 microl of 0.2 M NaCN was used, which did not interfere with the following labelling step. The quantification of the 32P-labelled dinucleotides was performed by phosphorimaging of spots after separation on TLC as well as by 32P-counting of fractions collected after separation by HPLC. The method was used to determine adduct levels in in vitro cisplatin-treated DNA and in DNA isolated from cisplatin-treated cultured cells, tumor xenografts from cisplatin-treated mice, and from white blood cells and (tumor) tissues from cisplatin-treated patients. The results show a significant correlation with the adduct levels as determined with atomic absorption spectroscopy (high levels) or with specific antibodies (low levels). This assay appears to be useful for the determination of low levels of Pt-adducts in small DNA samples as present in clinical specimens such as blood and tumor tissue, but also in buccal mucosal cells and fine needle aspirates. PMID- 9328175 TI - Cytochrome P450 mediated bioactivation of methyleugenol to 1' hydroxymethyleugenol in Fischer 344 rat and human liver microsomes. AB - Cytochrome P450 mediated metabolism of methyleugenol to the proximate carcinogen 1'-hydroxymethyleugenol has been investigated in vitro. Kinetic studies undertaken in liver microsomes from control male Fischer 344 rats revealed that this reaction is catalyzed by high affinity (Km of 74.9 +/- 9.0 microM, Vmax of 1.42 +/- 0.17 nmol/min/nmol P450) and low affinity (apparent Km several mM) enzymic components. Studies undertaken at low substrate concentration (20 microM) with microsomes from livers of rats treated with the enzyme inducers phenobarbital, dexamethasone, isosafrole and isoniazid indicated that a number of cytochrome P450 isozymes can catalyze the high affinity component. In control rat liver microsomes, 1'-hydroxylation of methyleugenol (assayed at 20 microM substrate) was inhibited significantly (P < 0.05) by diallylsulfide (40%), p nitrophenol (55%), tolbutamide (30%) and alpha-naphthoflavone (25%) but not by troleandomycin, furafylline, quinine or cimetidine. These results suggested that the reaction is catalyzed by CYP 2E1 and by another as yet unidentified isozyme(s) (most probably CYP 2C6), but not by CYP 3A, CYP 1A2, CYP 2D1 or CYP 2C11. Administration of methyleugenol (0-300 mg/kg/day for 5 days) to rats in vivo caused dose-dependent auto-induction of 1'-hydroxylation of methyleugenol in vitro which could be attributed to induction of various cytochrome P450 isozymes, including CYP 2B and CYP 1A2. Consequently, high dose rodent carcinogenicity studies are likely to over-estimate the risk to human health posed by methyleugenol. The rate of 1'-hydroxylation of methyleugenol in vitro in 13 human liver samples varied markedly (by 37-fold), with the highest activities being similar to the activity evident in control rat liver microsomes. This suggests that the risk posed by dietary ingestion of methyleugenol could vary markedly in the human population. PMID- 9328176 TI - Sensitivity of Escherichia coli (MutT) and human (MTH1) 8-oxo-dGTPases to in vitro inhibition by the carcinogenic metals, nickel(II), copper(II), cobalt(II) and cadmium(II). AB - The toxicity of Ni(II), Co(II) and Cu(II) in animals, and that of Cd(II) in cultured cells, has been associated with generation of the promutagenic lesion 8 oxo-7,8-dihydroguanine (8-oxoguanine) in DNA, among other effects. One possible source of this base may be 8-oxo-7,8-dihydro-2'-deoxyguanosine-5'-triphosphate (8 oxo-dGTP), a product of oxidative damage to the nucleotide pool, from which it is incorporated into DNA. To promote such incorporation, the metals would have to inhibit specific cellular 8-oxo-dGTPases that eliminate 8-oxo-dGTP from the nucleotide pool. The present study was designed to test such inhibition in vitro on 8-oxo-dGTPases from two different species, the human MTH1 protein and Escherichia coli MutT protein. In the presence of Mg(II), the natural activator of 8-oxo-dGTPases, all four metals were found to inhibit both enzymes. For MTH1, the IC50 values (+/- SE; n = 3-4) were 17 +/- 2 microM for Cu(II), 30 +/- 8 microM for Cd(II), 376 +/- 71 microM for Co(II) and 801 +/- 97 microM for Ni(II). For MutT, they were 60 +/- 6 microM for Cd(II), 102 +/- 8 microM for Cu(II), 1461 +/- 96 microM for Ni(II) and 8788 +/- 1003 microM for Co(II). Thus, Cu(II) and Cd(II) emerged as much stronger inhibitors than Ni(II) and Co(II), and MTH1 appeared to be generally more sensitive to metal inhibition than MutT. Interestingly, in the absence of Mg(II), the activity of the enzymes could be restored by Co(II) to 73% of that with Mg(II) alone for MutT, and 34% for MTH1, the other metals being much less or non-effective. The difference in sensitivity to metal inhibition between the two enzymes may reflect the differences in the amino acid ligands, especially the cysteine ligand, outside their evolutionarily conserved Mg(II)-binding active sites, which might indicate predominantly non competitive or uncompetitive mechanism of the inhibition. The overall results suggest that inhibition of 8-oxo-dGTPases may be involved in the mechanisms of induction of the 8-oxoguanine lesion in DNA by the metal ions studied, especially the non-redox-active Cd(II) cation. PMID- 9328177 TI - Metabolism of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) by human cytochrome P4501B1. AB - Cytochrome P4501B1 (CYP1B1) is the most recently identified member of the dioxin inducible CYP1 family. CYP1B1 is constitutively expressed in most human tissues, including colon and breast, and can activate numerous chemically diverse carcinogens. We evaluated the metabolism of the dietary heterocyclic amine carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) by microsomes from yeast expressing the human CYP1B1 protein. PhIP metabolites were analysed by HPLC with fluorescence and absorbance detection. We found that human CYP1B1 metabolizes PhIP to three products: N2-OH-PhIP, a mutagenic activation product; 4'-OH-PhIP, a detoxification product; and 2-OH-PhIP, the mutagenic potential of which is unknown. Metabolite identity was confirmed by co-elution with authentic standards and synchronous fluorescence spectroscopy. The identity of the 2-OH PhIP standard was additionally confirmed by mass spectrometry. Kinetic studies of the formation of N2-OH-PhIP, 4'-OH-PhIP and 2-OH-PhIP by CYP1B1 indicated apparent Km values of 5.7 +/- 1.3, 2.2 +/- 0.5 and 1.3 +/- 0.2 microM, respectively. Apparent turnover rates were 0.40 +/- 0.03, 0.93 +/- 0.02 and 0.04 +/- 0.00 nmol product/min nmol P450, respectively. At saturating levels of substrate, CYP1B1-mediated formation of the non-mutagenic metabolite 4'-OH-PhIP was favored two-fold over that of the mutagenic metabolite, N2-OH-PhIP and >10 fold over that of 2-OH-PhIP. The formation of N2-OH-PhIP, a potent mutagen implicated in the etiology of human colon and breast cancer, indicates that CYP1B1 may play an important role in PhIP-mediated carcinogenesis. PMID- 9328178 TI - Chorionic gonadotropin inhibits rat mammary carcinogenesis through activation of programmed cell death. AB - Human chorionic gonadotropin (hCG) inhibits the progression of 7,12 dimethylbenz[a]anthracene (DMBA) induced mammary carcinomas. In order to determine whether this phenomenon was mediated by induction of programmed cell death or apoptosis, 45-day-old virgin Sprague-Dawley rats received 8 mg DMBA/100 g body weight; 20 days later they were injected daily with 100 IU hCG for 40 days (DMBA + hCG group). Age-matched untreated, hCG- and DMBA + saline treated rats were used as controls. Tissues were collected at the time of DMBA administration and at 5, 10, 20 and 40 days of hCG injection. RNA from mammary glands, adenocarcinomas and ovaries was probed for transforming growth factors (TGF) alpha and beta, and the apoptotic genes TRPM2, ICE, bcl2, bcl-XL, bcl-XS, p53 and c-myc. The mammary glands of hCG-treated animals with or without DMBA exhibited elevated expression of TRPM2, ICE, bcl-XS, c-myc and p53; and elevation in the apoptotic index. Mammary adenocarcinomas developed in those animals treated with hCG showed an elevation in the expression of p53, c-myc and ICE genes in comparison with the levels detected in the adenocarcinomas developed by the animals treated with DMBA alone. No significant alterations in the expression of any of the genes tested was observed in ovarian RNAs. These results led us to conclude that hCG induces programmed cell death in the mammary gland initiated in the carcinogenic process, that this process is p53 dependent, and is modulated by c-myc expression. Our data also indicate the possibility that a cell death program dependent on the bcl2 family exists, because of the potential involvement of p53, bcl-XS and Bax in apoptosis. This additional mechanism of tumor inhibition makes hCG treatment a useful approach for the prevention and therapy of breast cancer. PMID- 9328179 TI - Inhibition of 2-nitropropane-induced rat liver DNA and RNA damage by benzyl selenocyanate. AB - We observed that pretreatment of male F344 rats with benzyl selenocyanate, a versatile organoselenium chemopreventive agent in several animal model systems, decreases the levels of DNA and RNA modifications produced in the liver by the hepatocarcinogen 2-nitropropane. To clarify the mechanisms involved, we pretreated male F344 rats with either benzyl selenocyanate, its sulfur analog benzyl thiocyanate, phenobarbital or cobalt protoporphyrin IX; the latter is a depletor of P450. We then determined (1) the ability of liver microsomes to denitrify 2-nitropropane, (2) effects on 2-nitropropane-induced liver DNA and RNA modifications and (3) amount of nitrate excreted in rat urine following administration of the carcinogen. Pretreatment with benzyl selenocyanate or phenobarbital increased the denitrification activity of liver microsomes by 217 and 765%, respectively, increased liver P4502B1 by 31- and 435-fold, respectively, decreased the levels of 2-nitropropane-induced modifications in liver DNA (29-70% and 17-30%, respectively) and RNA (67-85% and 30-50%, respectively), and increased the 24-h urinary excretion of nitrate by 157 and 209%, respectively. Pretreatment with benzyl thiocyanate had no significant effect on any of these parameters. Pretreatment with cobalt protoporphyrin IX decreased liver P4502B 1 by 87%, decreased the denitrification activity of liver microsomes by 76%, decreased the 24 h urinary excretion of nitrate by 88.5%, but increased the extent of 2-nitropropane-induced liver nucleic acid modifications by 17-67%. These results indicate that the metabolic sequence from 2-nitropropane to the reactive species causing DNA and RNA modifications does not involve the removal of the nitro group. Moreover, they suggest that benzyl selenocyanate inhibits 2-NP-induced liver nucleic acid modifications in part by increasing its detoxication through induction of denitrification, although it is evident that other mechanisms must also be involved. PMID- 9328180 TI - Effects of protein kinase C activators on phorbol ester-sensitive and -resistant EL4 thymoma cells. AB - Phorbol ester-sensitive EL4 murine thymoma cells respond to phorbol 12-myristate 13-acetate with activation of ERK mitogen-activated protein kinases, synthesis of interleukin-2, and death, whereas phorbol ester-resistant variants of this cell line do not exhibit these responses. Additional aspects of the resistant phenotype were examined, using a newly-established resistant cell line. Phorbol ester induced morphological changes, ERK activation, calcium-dependent activation of the c-Jun N-terminal kinase (JNK), interleukin-2 synthesis, and growth inhibition in sensitive but not resistant cells. A series of protein kinase C activators caused membrane translocation of protein kinase C's (PKCs) alpha, eta, and theta in both cell lines. While PKC eta was expressed at higher levels in sensitive than in resistant cells, overexpression of PKC eta did not restore phorbol ester-induced ERK activation to resistant cells. In sensitive cells, PKC activators had similar effects on cell viability and ERK activation, but differed in their abilities to induce JNK activation and interleukin-2 synthesis. PD 098059, an inhibitor of the mitogen activated protein (MAP)/ERK kinase kinase MEK, partially inhibited ERK activation and completely blocked phorbol ester induced cell death in sensitive cells. Thus MEK and/or ERK activation, but not JNK activation or interleukin-2 synthesis, appears to be required for phorbol ester-induced toxicity. Alterations in phorbol ester response pathways, rather than altered expression of PKC isoforms, appear to confer phorbol ester resistance to EL4 cells. PMID- 9328181 TI - Benzo[a]pyrene at an environmentally relevant dose is slowly absorbed by, and extensively metabolized in, tracheal epithelium. AB - While tobacco smoke has been conclusively identified as a lung carcinogen, there is much debate over which smoke constituent(s) are primarily responsible for its carcinogenicity. Previous studies in our laboratory suggested that highly lipophilic carcinogens are slowly absorbed in the thicker epithelium of the conducting airways, potentially allowing for substantial local metabolism. The bioactivation of polycyclic aromatic hydrocarbons in airway epithelium may, hence, be important in tobacco smoke-induced carcinogenesis. In the present study, the hypothesis of slow absorption and substantial local metabolic activation of highly lipophilic carcinogen in airway epithelium was tested in dogs. A single dose of tritiated benzo[a]pyrene (BaP) dissolved in a saline/phospholipid suspension was instilled in the trachea, just anterior to the carina. At intervals of a few minutes up to 30 min over a 3-h period, blood samples were drawn from the azygous vein, which drains the area around the point of instillation, and from the systemic circulation. Tissue samples were taken at the end of the experiment. The concentration of BaP with depth into the tracheal mucosa was determined with autoradiography. BaP was slowly absorbed into the trachea with a half-time of approximately 73 min, which is consistent with diffusion-limited passage through the epithelium and lead to local doses in the tracheal epithelium that were more than a 1000-fold those of other tissues. The long retention of BaP in the epithelium provided the local metabolizing enzymes with high substrate levels over a long period, resulting in extensive metabolism. At 3 h after the exposure, 23% of the BaP-equivalent activity remained in the tracheal mucosa. Of this fraction, 13% was parent compound, 28% was organic extractable, 31% was water-soluble, and 28-7% of the instilled dose was bound to tracheal tissues. These results explain the tendency of highly lipophilic carcinogens, such as BaP, to induce tumors at the site of entry and, furthermore, indicate that the highly lipophilic components of tobacco smoke and polluted air may be the most important contributors to lung tumors of the conducting airways. PMID- 9328182 TI - Problems in the measurement of 8-oxoguanine in human DNA. Report of a workshop, DNA oxidation, held in Aberdeen, UK, 19-21 January, 1997. AB - Oxidative DNA damage is widely believed to play a role in cancer aetiology. It is therefore important to be able to assess it, both as an index of cancer risk, and in experiments to test agents with a potential to reduce oxidative damage, such as dietary antioxidants. However, there is an alarming discordance in estimates of concentrations of oxidative damage in human DNA, largely attributable to the kind of method used to measure it. A meeting was held recently at the Rowett Research Institute in Aberdeen to address this problem. PMID- 9328183 TI - Molecular cloning and characterization of the promoter of the human N methylpurine-DNA glycosylase (MPG) gene. AB - The promoter region of the human N-methylpurine-DNA glycosylase (MPG) gene was cloned and characterized. The cloned segment contains two first exons that were earlier identified and named exons 1a and 1b. These were found to be separated by approximately 800 bp. The minimal promoter region was identified upstream to the distal exon 1a, by transient transfection, and no promoter activity was found in the region in between exons 1a and 1b, suggesting that transcription starts at a single site which is then processed to generate mRNAs of the isoforms. The promoter sequence is G and C rich and contains neither TATA box, nor apparent CAAT sequences, although a partially matched CAAT sequence was identified just downstream to the minimal promoter. PMID- 9328184 TI - Expression of differential nitric oxide synthase isoforms in human normal gastric mucosa and gastric cancer tissue. AB - The present study investigated the expression and distribution of three isoforms of nitric oxide synthase (NOS) in different anatomical regions of the human stomach and in gastric neoplastic tissues by immunohistochemistry using specific antibodies. Intracellular localization of individual isoenzymes of NOS was detected in normal gastric mucosa. Gastric cancer tissues had a marked reduction of all three NOS isoforms expression. The expression of the endothelial NOS, neuronal NOS and inducible NOS in the tumor tissue was significantly lower than in normal gastric mucosa (P = 0.01, P = 0.02, P < 0.01, respectively). In the tumor tissue the expression of inducible NOS was significantly lower than the expression of both constitutive forms of NOS (P < 0.01). There was a tendency to higher expression of both constitutive forms of NOS in earlier stages T2 of the tumor compared to advanced T4 tumor. In contrast, the expression of inducible NOS was higher than in the advanced T4 tumor than in the earlier stages T2 of the tumor. The mapping of the expression of endothelial NOS, neuronal NOS and inducible NOS in human stomach showed higher expression of NOS isoforms in the distal third than in the proximal third of the stomach (P = 0.03, P = 0.04, P = 0.01, respectively). We conclude that there is greater expression of NOS in the stomach corpus and in antrum than in the proximal third of the normal human stomach mirroring the anatomical predilection of common pathological changes in this part of the human stomach. Furthermore, there was loss of the expression of individual isoenzymes in gastric neoplasms. PMID- 9328185 TI - Consumption of vegetables reduces genetic damage in humans: first results of a human intervention trial with carotenoid-rich foods. AB - A human intervention study with vegetable products has been performed in twenty three healthy, non smoking males aged 27-40. It was the aim of the study to assess whether consumption of vegetables containing different carotenoids could protect against DNA damage and oxidative DNA damage. The subjects consumed their normal diets, but abstained from vegetables high in carotenoids throughout the study period. After a 2 week depletion period, they received daily 330 ml tomato juice with 40 mg lycopene (weeks 3 and 4), 330 ml carrot juice with 22.3 mg beta carotene and 15.7 mg alpha-carotene (weeks 5 and 6), and 10 g dried spinach powder (in water or milk) with 11.3 mg lutein (weeks 7 and 8). Blood was collected weekly and DNA damage was detected in peripheral blood lymphocytes with the 'COMET' assay. Oxidised DNA bases were detected by including an incubation step with endonuclease III. The supplementation of the diet with tomato, carrot or spinach products resulted in a significant decrease in endogenous levels of strand breaks in lymphocyte DNA. Oxidative base damage was significantly reduced during the carrot juice intervention. These findings support the hypothesis that carotenoid containing plant products exert a cancer-protective effect via a decrease in oxidative and other damage to DNA in humans. PMID- 9328186 TI - Absolute configuration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol formed metabolically from 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is an important metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone (NNK). Using the chiral derivatizing agent, (R)-(+)-alpha-methylbenzyl isocyanate [(R)-(+)-MBIC], previous work has shown that the enantiomeric ratio of metabolically formed NNAL and its glucuronide derivative may be species dependent. However, the absolute configuration of such NNAL has not been previously reported. Synthetically prepared racemic NNAL was converted to diastereomeric esters by reaction with (R)-(+)- and (S)-(-)-alpha-methoxy-alpha (trifluoromethyl)phenylacetic acid (MTPA) chloride (Mosher's reagent) and the products were characterized by 1H-NMR. Based on chemical shift data, the absolute configuration of NNAL in each diastereomeric ester was assigned. Hydrolysis of (R)-NNAL-(R)-MTPA gave (R)-NNAL. This was converted to the corresponding carbamate by reaction with (R)-(+)-alpha-MBIC and the absolute configurations of the diastereomeric carbamates formed by reaction of (R)- and (S)-NNAL with (R) (+)-MBIC were thereby assigned. Conversion of metabolically produced NNAL to the same carbamates allowed us to assign the NNAL formed from NNK by rat liver microsomes as (R)-NNAL. The major and minor NNAL-glucuronide diastereomers found in the urine of patas monkeys and humans exposed to NNK were similarly assigned; they were formed from (R)-NNAL and (S)-NNAL, respectively. PMID- 9328187 TI - Effects of 1,4-phenylenebis(methylene)selenocyanate and selenomethionine on 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced tumorigenesis in A/J mouse lung. AB - We reported earlier that continuous feeding of 1,4 phenylenebis(methylene)selenocyanate (p-XSC) inhibited lung tumor induction by the tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the A/J mouse (El-Bayoumy et al., Carcinogenesis, 14, 1111-1113, 1993). The present investigation was designed to determine whether p-XSC inhibits pulmonary neoplasia induced by NNK in female A/J mice during the initiation phase of carcinogenesis or during the post-initiation phase. The naturally occurring selenomethionine was also included in this study. Doses higher than 4 p.p.m. of selenomethionine can induce toxic effects, therefore, dietary supplementation of this compound was selected at a dose level of 3.75 p.p.m. However, we were able to give p-XSC at selenium levels of 7.5 and 15 p.p.m., as mice can tolerate such doses in this form without any adverse effects. NNK was given by a single i.p. injection at dose of 10 micromol in 0.1 ml of saline. Selenomethionine did not show chemopreventive activity when administered in either phase of tumorigenesis. In contrast, p-XSC significantly reduced lung tumor multiplicity regardless of whether it was given during the initiation phase of tumorigenesis (P = 0.0009 at both levels of selenium) or post-initiation (P = 0.0009 at 15 p.p.m. and P = 0.036 for 7.5 p.p.m.). This is the first report describing that the synthetic organoselenium compound, p-XSC, can effectively block and suppress chemically (NNK)-induced lung tumor development in mice. PMID- 9328188 TI - Re: Feigelson, H.S. and Henderson, B.E. (1996) Estrogens and breast cancer. Carcinogenesis, 17, 2279-2284. PMID- 9328189 TI - To supplement or not to supplement, that is the question. PMID- 9328190 TI - Parental occupation, occupational exposure to solvents and polycyclic aromatic hydrocarbons and risk of childhood brain tumors (Italy, France, Spain) AB - The role of parental occupational exposure in childhood brain tumors was investigated in a population-based case-control study grouping 251 cases and 601 controls from three European centers: Milan (Italy), Paris (France), and Valencia (Spain). Parental occupational exposure to solvents and polycyclic aromatic hydrocarbons (PAH) during the five-year period before birth was estimated using a job-exposure matrix developed earlier in the same countries. Odds ratios (OR) of brain tumors for each occupation and occupational exposure were estimated by logistic regression, adjusting for child's age, gender, exposure to tobacco smoke and ionizing radiation, mother's age and years of schooling, and center. The risk of childhood brain tumors rose when fathers worked in agriculture (OR = 2.2, 95 percent confidence interval [CI] = 1.0-4.7) and motor-vehicle-related occupations. In the latter group, the risk increased for primitive neuroectodermal tumors in particular (OR = 2.7, CI = 1.1-6.6). Astroglial tumors were more frequent among children of mothers in health services (OR = 2.2, CI = 1.0-4.9). Paternal exposure to PAHs was associated with an increased, but not dose-related, risk of primitive neuroectodermal tumors (OR = 2.0, CI = 1.0-4.0), and maternal exposure to solvents at a high level was associated with an increased risk of both astroglial (OR = 2.3, CI = 0.9-5.8) and primitive neuroectodermal tumors (OR = 3.2, CI = 1.0-10.3). PMID- 9328191 TI - The cervical cancer screening program in Mexico: problems with access and coverage. AB - A cross-sectional study was carried out in two geographic regions of Mexico - Oaxaca (rural area) and Mexico City (urban area) - to determine the main factors for predicting participation in Cervical Cytology Screening Programs (CCSP), in populations with high mortality due to cervical cancer. We included 4,208 women aged between 15 and 49 years, randomly selected through a national household sample frame. Knowledge of what the Pap test is used for strongly predisposes use of CCSP in Mexico City (odds ratio [OR] = 46.1, 95 percent confidence interval [CI] = 33.1-64.1) and Oaxaca state (OR = 61.5, CI = 42.0-89.9), as well as high socioeconomic level (Mexico: OR = 2.0, CI = 1.1-7.6; Oaxaca: OR = 4.1, CI = 3.1 5.3), high education level (Mexico: OR = 3.6, CI = 1.5-8.8; Oaxaca: OR = 5.3, CI = 2.8-10.0), and access to social security (Mexico: OR = 1.7, CI = 1.4-2.2; Oaxaca: OR = 2.2, CI = 1.8-2.7). Low coverage of the CCSP is confirmed as an important problem in Mexico. PMID- 9328192 TI - Cancer incidence in the Hmong of Central California, United States, 1987-94. AB - The Hmong are an ethnic minority in China, some of whom migrated to the mountainous areas of North Vietnam, Laos, and Thailand in the 19th and 20th centuries. Because of their support for the United States during the Vietnam war, many Laotian Hmong fled to Thailand and eventually were re-settled in the US after the end of that conflict. Approximately 100,000 Hmong currently live in the US, of whom about half reside in the Central Valley of California. The purpose of this study was to measure cancer incidence in this unique new immigrant population. Using the resources of the Cancer Registry of Central California (CRCC), a population-based cancer registry, cancer incidence in the Hmong was evaluated by calculating age-adjusted incidence rates as well as by calculating proportional incidence ratios. Compared with all races combined, elevated rates of cancer in the Hmong were observed for the following sites: nasopharynx, stomach, liver, pancreas, leukemia, and non-Hodgkin's lymphoma. Cervical cancer incidence overall was elevated, but more noteworthy, invasive cervix cancer rates were much higher than expected. Lower cancer rates were found for breast, prostate, and colorectal cancer. Hmong also experienced advanced stage and grade of disease at diagnosis for many cancer sites in addition to cervical cancer, which may be explained by cultural factors, including avoidance of Western medical care and low rates of participation in screening programs. This population should be followed closely and monitored for patterns of cancer incidence in the future since it provides a natural laboratory for studies of cancer etiology as this population gradually becomes acculturated to the Western lifestyle. PMID- 9328193 TI - Dietary relationships with early onset (under age 45) breast cancer in a case control study in the United States: influence of chemotherapy treatment. AB - Methodologic investigations have addressed selection and recall bias in case control studies of diet and breast cancer, whereas the effect of disease progression and medical treatment on estimates of dietary intake has been largely overlooked. In a multicenter, population-based case-control study of breast cancer in the United States, 1,588 newly diagnosed cases and 1,451 controls completed a self-administered food-frequency questionnaire. Initial evaluation suggested increased risk related to high intakes of calories, carbohydrates, fat, and protein. All nutrient associations were diminished after adjustment for calories. Evaluation by stage of disease revealed no relation of calories to risk among women with in situ disease, but elevated risks among women with localized (odds ratio [OR] = 1.33, 95 percent confidence interval [CI] = 1.0-1.7 highest cflowest quartile) or regional and distant disease (OR = 1.79, CI = 1.3-2.4). Further evaluation showed that the increased risk associated with calories was restricted to cases who reported having been treated with chemotherapy (OR = 1.66, CI = 1.3-2.1). A gradient of increasing risk with time interval from diagnosis to interview suggested the chemotherapy regimen itself and not necessarily characteristics of tumors requiring this treatment was responsible for the observed increased risk. These results indicate that epidemiologic studies of diet and breast cancer, particularly among young women, should evaluate possible bias related to post-diagnosis influences. PMID- 9328194 TI - Marijuana use and cancer incidence (California, United States). AB - The purpose of this retrospective cohort study was to examine the relationship of marijuana use to cancer incidence. The study population consisted of 64,855 examinees in the Kaiser Permanente multiphasic health checkup in San Francisco and Oakland (California, United States), between 1979-85, aged 15 to 49 years, who completed self-administered questionnaires about smoking habits, including marijuana use. Follow-up for cancer incidence was conducted through 1993 (mean length 8.6 years). Compared with nonusers/experimenters (lifetime use of less than seven times), ever- and current use of marijuana were not associated with increased risk of cancer of all sites (relative risk [RR] = 0.9, 95 percent confidence interval [CI] = 0.7-12 for ever-use in men; RR = 1.0, CI = 0.8-1.1 in women) in analyses adjusted for sociodemographic factors, cigarette smoking, and alcohol use. Marijuana use also was not associated with tobacco-related cancers or with cancer of the following sites: colorectal, lung, melanoma, prostate, breast, cervix. Among nonsmokers of tobacco cigarettes, ever having used marijuana was associated with increased risk of prostate cancer (RR = 3.1, CI = 1.0-9.5) and nearly significantly increased risk of cervical cancer (RR = 1.4, CI = 1.0-2.1). We conclude that, in this relatively young study cohort, marijuana use and cancer were not associated in overall analyses, but that associations in nonsmokers of tobacco cigarettes suggested that marijuana use might affect certain site-specific cancer risks. PMID- 9328195 TI - Tobacco, alcohol use, and risks of laryngeal and lung cancer by subsite and histologic type in Turkey. AB - Effects of tobacco smoking and alcohol use on risks of cancers of the larynx and lung have been evaluated extensively in industrialized countries. Few studies on the effect of these risk factors have been reported from developing countries. We conducted a case-control study to evaluate risks of laryngeal and lung cancers in men by subsite and cell type in relation to smoking and alcohol drinking in Turkey, a country where smoking and alcohol consumption patterns are different from those in industrialized countries. We identified 832 laryngeal and 1,210 lung cancer cases and 829 controls with information on smoking and alcohol use (amount and duration) and histologic cell type from an oncology treatment center of a Social Security Agency hospital in Istanbul, Turkey, admitted between 1979 and 1984. Both laryngeal and lung cancer showed significant associations with smoking and alcohol drinking, but no monotonic dose-response was obtained for alcohol drinking. Among smokers, the highest risks were observed in the supraglottis region of the larynx (odds ratio [OR] = 4.1) after adjustment for age and alcohol use. Among alcohol drinkers, the highest risks were observed in the glottis region of the larynx (OR = 1.7) after adjustment for age and smoking. In the analysis by the cell type of lung cancer among ever-smokers, small cell type showed the highest risk (OR = 5.4), while it showed no association with alcohol drinking. Cumulative cigarette use (pack-years) and number of cigarettes per day showed stronger associations than years smoked for both cancer sites. The relative risks of joint exposure to smoking and alcohol were 12.2 for laryngeal cancer and 14.1 for lung cancer among heavy smokers and heavy alcohol drinkers. This study provides epidemiologic evidence from Turkey that smoking and alcohol use are associated with risks of cancers of the larynx and lung. PMID- 9328196 TI - Bladder cancer and drinking water: a population-based case-control study in Washington County, Maryland (United States). AB - A population-based case-control study was conducted in Washington County, Maryland (United States) to explore the association between incident bladder cancer and exposure to drinking water from chlorinated surface sources. Cancer cases were White residents, enumerated in a 1975 county census and reported to the Washington County Cancer Registry (n = 294) between 1975 and 1992. White controls, frequency matched by age (+/- 5 years) and gender, were selected randomly from the census (n = 2,326). Households receiving municipal water, which generally derived from chlorinated surface waters, were treated as having 'high exposure' and all others, as 'low exposure.' Duration of exposure to type of drinking water was based on length of residence in the census household prior to 1975. Odds ratios (OR) were calculated using logistic regression methods, adjusting for age, gender, tobacco use, and urbanicity. Bladder cancer risk was associated weakly in the general population with duration of exposure to municipal water. The association was limited to those who had smoked cigarettes. In ever-smokers compared with never-smokers with low exposure, the adjusted ORs for bladder cancer risk with increasing exposure were 1.3, 1.4, 1.4, 1.7, 2.2, 2.8, respectively, for 0, 1-10, 11-20, 21-30, 31-40, > 40 years' exposure duration. The ORs in smokers were not diminished after adjusting for smoking history and intensity. PMID- 9328197 TI - Incidence and mortality of neuroblastoma in Canada compared with other childhood cancers. AB - The incidence and mortality of neuroblastoma was reviewed in the general context of childhood cancer in Canada for the periods 1982-86 and 1987-91. This was done to complement the preliminary work of the Quebec Neuroblastoma Screening Project that is studying the impact of screening North American infants for the preclinical detection of neuroblastoma on population-based mortality. Annual age standardized incidence rates for all childhood cancer in Canada appear to have declined slightly (nonsignificantly) from 155.1 to 150.8 per million, between 1982-86 and 1987-91; the rates for neuroblastoma were stable between the two five year periods (11.8 per million in 1982-86 and 11.4 per million in 1987-91). With respect to mortality, the age-standardized rates for childhood cancer in Canada have shown a declining trend between the first and second halves of the decade, from 43.4 to 34.7 per million, while the rates for neuroblastoma have not changed (4.4 and 4.2 per million). The age-specific distributions of incident cancers indicate that neuroblastoma accounts for the greatest proportion of all cancers in children less than one year of age. Similarly, neuroblastoma is the leading cause of cancer deaths in children aged one to four years. Theoretically, infants less than one year of age could benefit most from effective preventive interventions, treatment, and research. PMID- 9328198 TI - International incidence rates of invasive cervical cancer after introduction of cytological screening. AB - Because Pap-smear screening can detect pre-invasive cervical cancer, such screening can markedly reduce the occurrence of invasive cancer. However, its impact in different populations is uncertain. This study compares the changes in cervical cancer incidence at different ages after the introduction of screening in different populations, and addresses the impact of organized and opportunistic smear taking. We identified 17 cancer registries large enough and existing long enough to analyze screening effects. For each registry, we calculated the relative reduction in age-specific incidence rates and in incidence rates age standardized to the world population after the introduction of cytologic screening. In 11 of the 17 populations, age-standardized incidence rates declined markedly from 27 percent in Norway and to 77 percent in Finland. Age-specific declines were confined to women aged 30 to 70 years old with a nadir around ages 40 to 55. In six other populations, age-standardized incidence rates declined less than 25 percent, an amount too small to provide unambiguous evidence of a screening effect. In several populations, cytologic screening had a more pronounced effect than is generally recognized. Because age-specific declines in cervical cancer incidence rates were strikingly similar in populations with widely different screening practices, organized screening may not be markedly superior to opportunistic screening. The reduction in reported cancer incidence because of screening is smaller in younger and older women. PMID- 9328199 TI - Second primary cancers in breast cancer patients in Slovenia. AB - Data from the Cancer Registry of Slovenia were used in a cohort study to determine whether the incidence of second primary cancers in patients with first primary breast cancer differs from the incidence expected in the general population. Special interest was given to long-term survivors. The expected numbers of second primary cancers were calculated by multiplying the number of appropriate person-years at risk by the corresponding age- and calendar-period specific cancer incidence rates for women in Slovenia. The risk of a second primary cancer was expressed as the standardized incidence ratio (SIR). Of the 8,917 patients newly diagnosed in the period 1961-85 and followed-up to the end of 1994, 547 (6.2 percent) developed second primary cancers, whereas 410 (4.7 percent) were expected (SIR = 1.3, 95 percent confidence interval [CI] = 1.2 1.4). The risk was higher among younger patients. In long-term survivors, the risk was increased significantly for second primary cancer of the breast (SIR = 1.4, CI = 1.1-1.7), lung cancer (SIR = 1.6, CI = 1.1-2.3), melanoma (SIR = 2.7, CI = 1.5-4.4) and non-melanoma skin cancers(SIR = 2.0, CI = 1.6-2.4), corpus uteri cancer(SIR = 1.6, CI = 1.2-2.1), ovarian cancer(SIR = 2.3, CI = 1.7-3.0), and thyroid cancer (SIR = 2.5, CI = 1.2-4.6). Our results confirm the findings of several cohort studies carried out in Europe, the United States, and Japan, indicating that breast cancer patients should be monitored carefully for the occurrence of second primary cancers. PMID- 9328200 TI - Factors determining acceptability of mammography in an Asian population: a study among women in Singapore. AB - Population-based mammographic screening has been shown to be effective in reducing breast cancer mortality in the West. In Singapore, a project carried out to determine the effectiveness of implementing such a program locally invited 28,000 women between the ages of 50 and 64 years for mammography. The current study, which was part of this larger project, was intended to determine factors contributing to the acceptance of mammographic screening among women in Singapore. A questionnaire was administered in-person to 300 attenders and 260 non-attenders. The respondents were compared with respect to basic demographic characteristics, previous preventive behavior, informal social support, and attitudes towards early detection. We found that screening attenders were more likely to be Chinese than Malays (14 percent of the population) or Indians (seven percent), and to be working outside the home (adjusted odds ratio [OR]) = 4.5, 95 percent confidence interval [CI] = 2.6-7.9). A greater proportion of attenders had a history of other screening tests such as the Pap smear (OR = 4.7, CI = 2.6 8.7 for recent smear compared with never having had a smear). They were also more likely to indicate a sense of personal susceptibility to cancer, but did not differ from non-attenders in terms of believing in cancer prevention, or of preferring to be told if they did have cancer. The strongest independent predictor of attendance, however, was encouragement by her spouse or family member. For women in this population to be persuaded effectively to participate in mammographic screening, it would be important to convince family members of the benefits of the test. At the same time, education targeted specifically at women of the appropriate age group should address the issue of the personal relevance of screening for breast cancer. PMID- 9328201 TI - Case-control study of colorectal carcinoma in situ and cancer in relation to cigarette smoking and alcohol use (Japan). AB - A case-control study was performed in order to examine the relation of cigarette smoking and alcohol use to colorectal carcinoma in situ and cancer, separately. Study subjects consisted of 129 colorectal carcinoma in situ cases, 66 colorectal cancer cases, and 390 controls recruited from health check-up examinees in Tokyo from January 1991 to March 1993. Smoking status and alcohol habit were ascertained from a self-administered questionnaire. Both cumulative cigarette smoking and current smoking status were associated significantly with an increased risk of carcinoma in situ. A statistically nonsignificant increase in the risk of colorectal cancer was noted only among those with the heaviest exposure category of these measures of smoking. The cumulative exposure to cigarette smoking within recent 20 years was associated significantly with an increased risk of carcinoma in situ, whereas smoking until 20 years before the diagnosis was associated significantly with an increase of cancer risk. A significant and positive association was observed between cumulative alcohol drinking and colorectal cancer. These findings suggest that cigarette smoking may act as an initiator in colorectal carcinogenesis and also provide weak evidence that alcohol drinking is related to an increased risk of colorectal cancer. PMID- 9328202 TI - Vitamin supplements and cancer risk: the epidemiologic evidence. AB - This report reviews published epidemiologic research on the associations of vitamin and mineral supplementation with cancer risk. Although the literature on nutrition and cancer is vast, few reports to date have addressed supplemental nutrients directly (seven clinical trials, 16 cohort, and 36 case-control studies). These studies offer insight into effects of nutrients that are distinguishable from effects of other biologically active compounds in foods. Randomized clinical trials have not shown significant protective effects of beta carotene, but have found protective effects of: alpha-tocopherol against prostate cancer; mixtures of retinol/zinc and beta-carotene/alpha-tocopherol/selenium against stomach cancer; and selenium against total, lung, and prostate cancers. Cohort studies provide little evidence that vitamin supplements are associated with cancer. Case-control studies have reported an inverse association between bladder cancer and vitamin C; oral/pharyngeal cancer and several supplemental vitamins; and several cancers and vitamin E. A randomized clinical trial, a cohort study, and a case-control study have all found inverse associations between colon cancer and vitamin E. Overall, there is modest evidence for protective effects of nutrients from supplements against several cancers. Future studies of supplement use and cancer appear warranted; however, methodologic problems that impair ability to assess supplement use and statistical modeling of the relation between cancer risk and supplement use need attention. PMID- 9328203 TI - Is Mycoplasma Pneumonia associated with childhood acute lymphoblastic leukemia? AB - Acute lymphoblastic leukemia (ALL) in the childhood peak may be a rare response to delayed first exposure to one or more common infectious agent(s). Mycoplasma Pneumonia has the appropriate socioeconomic correlates and clinical symptoms and the hypothesis that delayed first exposure to it may contribute to ALL is considered. Counts of positive reports of M Pneumonia from disease surveillance data for England and Wales (United Kindom) for 1975-92 have been taken as proxies for community burden of infection. Variation by months of birth (cohort) and diagnosis (period) of incidence of ALL in children born and diagnosed 1975-92 are compared with predictions. When periods were classified by mean M Pneumonia count rate in the nine preceding months, standardized morbidity ratios (SMR) for the highest and lowest 20 percent were 108 and 89 (rate ratio [RR] = 1.2, 95 percent confidence interval [CI] = 1.1-1.4). SMRs for cohorts with highest and lowest predicted risk (i.e., lowest and highest M Pneumonia count rate around birth and during infancy) were 110 and 97 (RR = 1.1, CI = 1.0-1.3). The trend for period was most marked in the cohorts with low opportunity for exposure when young. This ecologic analysis provides preliminary support for the hypothesis. PMID- 9328204 TI - Neurotransmitters activate the human estrogen receptor in a neuroblastoma cell line. AB - The human neuroblastoma cell line SK-N-SH has been used as a model system to study the interactions of the human estrogen receptor (hER) with neurotransmitters. We have successfully transfected these cells using an adenoviral delivery system and have reconstituted ligand-dependent responses to estradiol and ligand-independent responses to a series of dopamine D1 receptor agonists. The full agonist for the D1 receptor, SKF 82958, shows a robust activation of hER, comparable to that induced by estradiol. This activation is blocked by the protein kinase A inhibitor H-89, is mimicked by forskolin, and is therefore thought to be mediated in part through the cAMP/protein kinase A pathway. We have examined deletion mutants of hER for activation by SKF 82958 and find that both its transactivation domains, AF-1 and AF-2, must cooperate to impart the full response to the agonist. Significantly, an agonist of the muscarinic acetylcholine receptor, carbachol, though not active by itself, synergistically activates hER in conjunction with suboptimal doses of SKF 82958. This is the first reported instance of two neurotransmitters synergizing to activate a member of the nuclear receptor superfamily, and might predict a role for multiple neural inputs modulating the effects of these receptors in the central nervous system. PMID- 9328205 TI - Both estradiol and tamoxifen decrease proliferation and invasiveness of cancer cells transfected with a mutated estrogen receptor. AB - Previous studies have shown that, after wild-type estrogen receptor (ER) transfection in ER-negative breast cancer cells, estradiol but not tamoxifen prevents growth, invasiveness and metastasis of these cells in mice. Because an ER mutation at position 400 converts the triphenylethylene antiestrogen, OH tamoxifen into a full estrogen agonist, we transfected this mutated form of human ER in an ER-negative rat cancer cell line. This was aimed at inducing an inhibitory, estrogen-like response of tamoxifen in these cells. In two stable ER positive transfectants, OH-tamoxifen inhibited cell growth and invasiveness in vitro as efficiently as estradiol. The pure antiestrogen, ICI 164,384, was not agonistic alone and antagonized estrogen action. In contrast, the three compounds were ineffective in control mock-transfected cells. When injected into ovariectomized nude mice, ER-negative mock-transfected cells formed tumours which were significantly stimulated by estradiol and inhibited by tamoxifen treatment. This indicates that estradiol and tamoxifen altered the growth of ER-negative tumours via a general effect on the host response. Surprisingly, the hormone responsiveness of ER-positive tumours developed from ER-transfected cells did not significantly differ from that of ER-negative (mock-transfected) tumours. We conclude that transfection of a mutated human estrogen receptor inhibited, through an estrogenic activity of tamoxifen, the growth and invasiveness of these cancer cells in vitro. However, the low expression of ER did not allowed us to obtain the same effect of tamoxifen in vivo. PMID- 9328206 TI - Wide variation in androgen receptor dysfunction in complete androgen insensitivity syndrome. AB - Androgen insensitivity syndrome (AIS) is a disorder of male sexual differentiation caused by mutations in the androgen receptor (AR) gene. The partial form (PAIS), associated with varying degrees of receptor dysfunction, presents with a range of undervirilization phenotypes. The complete form (CAIS) is characterized by normal female external appearance at birth. In these cases the receptor is often absent or inactive. However, cases have been described where the mutant receptor concerned has considerable residual activity in in vitro assays. Here we describe the effects of five mutations, Gly750Asp, Leu762Phe, Ala765Thr, Asp864Asn and Leu907Phe, identified in complete androgen insensitivity patients. In vitro assays of mutant androgen receptors expressed in a mammalian cell line showed that the Gly750Asp, Leu762Phe and Ala765Thr mutations cause almost complete loss of androgen-binding activity, suggesting that these residues are critical for ligand binding. However, receptors with Asp864Asn and Leu907Phe, although defective, were capable of considerable binding and transactivation activity. Given that some mutations identified in PAIS patients have a more severe effect on androgen receptor function than two CAIS mutations described here, these results provide further evidence that other factors, including genetic background, can have a significant impact on the phenotype associated with a particular AR mutation. PMID- 9328207 TI - Steroid sulphotransferase and 17beta-hydroxysteroid dehydrogenase activities in Ishikawa human endometrial adenocarcinoma cells. AB - The present studies concern sulphotransferase activities for estrogens and other steroids, and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activities for estrogens in Ishikawa endometrial adenocarcinoma cells. When physiological concentrations of various estrogens (estrone, estradiol, estriol) are incubated, most of the transformation product is the respective sulphate. The sulphotransferase activity is very rapid, and 2 h after incubation 70-95% are converted to the sulphated form. Sulphates are found exclusively in the culture medium, which suggests that as soon as the sulphate is biosynthesized it is secreted to the medium. Comparative data using neutral steroids (dehydroepiandrosterone, testosterone, and pregnenolone) show that sulphotransferase activity for these compounds is very limited. In another series of studies, 17beta-HSD activity was explored for the interconversion estrone estradiol. At low concentrations (5 x 10(-9)-5 x 10(-8) M), when estradiol (E2) is incubated, most of the unconjugated material remains as E2 in the cellular compartment, but at high concentrations (5 x 10(-7)-5 x 10(-6) M) a great proportion (70-80%) of the E2 is converted to estrone (E1). On the other hand, after incubation of E1 at all concentrations most remained as unchanged E1. It is suggested that, in Ishikawa cells, at very low concentrations of E1 or E2, sulphotransferases are predominant, but when this enzyme is saturated 17beta-HSD activity is orientated to the oxidative form. PMID- 9328208 TI - Glucocorticoid/oxysterol-induced DNA lysis in human leukemic cells. AB - Both glucocorticoids and oxysterols, steroids with quite different known transduction pathways, cause the death of lymphoid cells. Dual TUNEL/propidium iodide assays on sensitive human leukemic CEM-C7 clones treated with either steroid were clearly positive by 48 h, consistent with apoptosis. Both steroids evoked two distinctive types of DNA lysis: cleavage into large fragments of several different sizes and the classic "ladders", multiples of approximately 200 base pairs. Conventional gel electrophoresis showed that a small proportion of total DNA had undergone laddering 36-48 h after treatment with glucocorticoid or 24 h after oxysterol exposure. On field inversion gel electrophoresis of cellular DNA both steroids caused an increase in an array of large DNA fragments <50 kb in size. A 50 kb fragment appeared 36 h after treatment with either steroid, but only oxysterol treatment caused a significant increase in a 300 kb fragment. Oxysterol treatment did not result in DNA fragmentation in the resistant M10R5 subclone, which retained sensitivity to glucocorticoids. We conclude that glucocorticoids and oxysterols kill these cells with similar, but not identical, patterns of DNA lysis which occur just before or concomitant with the onset of cell death. PMID- 9328210 TI - Inhibition of rat alpha-reductases by finasteride: evidence for isozyme differences in the mechanism of inhibition. AB - The mechanism of inhibition of the rat types 1 and 2 5alpha-reductase by finasteride was investigated using recombinantly expressed enzymes. These studies revealed that finasteride is a potent, reversible inhibitor of the rat type 1 5alpha-reductase with Ki=10.2+/-1.3 nM. Finasteride is a potent inhibitor of the rat type 2; however, in this case the compound binds to the type 2 isozyme-NADPH complex to form a ternary complex with Ki=1.19+/-0.10 nM, which then rearranges to a high affinity complex (E:I) with a pseudo first order rate constant of 1.62+/-0.22 x 10(-3)/s. The second order rate constant is k3/Ki=1.37+/-0.31 x 10(6) M/s. Heat denaturation of the (type 2 enzyme:inhibitor) complex releases dihydrofinasteride and presumably the NADP+-adduct previously identified with the human 5alpha-reductases. The effects of finasteride were also studied in intact COS cells transiently expressing the rat types 1 and 2 5alpha-reductase. Results with whole cell assays confirm differences in mechanism of inhibition of rat types 1 and 2 5alpha-reductase by finasteride. PMID- 9328209 TI - The role of nitric oxide in the regulation of aldosterone synthesis by adrenal glomerulosa cells. AB - The role of nitric oxide (NO) in the regulation of aldosterone synthesis in the adrenal glomerulosa is not known. In this study, we observed that liberators of NO such as S-nitroso-N-acetyl-penicillamine (SNAP), sodium nitroprusside (Snp) and spermine nonoate (SNO) could significantly inhibit angiotensin II (AII) and ACTH-induced aldosterone synthesis in isolated rat and cultured human adrenal glomerulosa cells. To evaluate more precisely whether glomerulosa cells express NO synthase, we performed immunoblotting experiments with antibodies specific for the endothelial type ecNO synthase as well as the neuronal NO synthase. This revealed the presence of the ecNO synthase in rat adrenal capsules, in normal and in adenomatous human adrenal glomerulosa tissue, as well as in freshly dispersed rat adrenal glomerulosa cells. Furthermore, on immunohistochemical analysis, rat adrenal glomerulosa cell sections showed strongly positive staining for ecNO synthase. These results suggest that NO may be an important negative modulator of adrenal glomerulosa steroidogenesis. PMID- 9328211 TI - Regulation of glucocorticoid receptor by the tyrosine kinase inhibitor herbimycin A in the cytosolic fraction of primary cultured rat hepatocytes. AB - The participation of tyrosine kinase in the regulation of the glucocorticoid receptor (GR) was studied in primary cultured rat hepatocytes using the tyrosine kinase inhibitor herbimycin A. Herbimycin A decreased the number of high-affinity binding sites of glucocorticoids in the cytosolic fraction and increased the equilibrium dissociation constant (Kd). Western blot analysis revealed that it also decreased the amount of GR protein. On the other hand, cycloheximide did not affect the GR protein level. Although herbimycin A slightly increased the amount of GR protein in the nuclear fraction, the increase was much lower than that of its decrease in the cytosolic fraction. Therefore, the decrease of GR protein in the cytosolic fraction was not caused by the inhibition of GR protein synthesis nor the translocation of GR from cytosol to nuclei. As herbimycin A also suppressed the dexamethasone (Dex)-dependent induction of tyrosine aminotransferase (TAT) activity, the decrease of GR protein was followed by the suppression of the GR-mediated biological response. These findings indicate that tyrosine kinase is necessary for the maintenance of the level of GR protein and its affinity of binding sites in the cytosolic fraction. Our results also suggested that the increase of GR protein stability is the most probable explanation for the maintenance of its level. PMID- 9328212 TI - Aromatase inhibition in JAr choriocarcinoma cells by 7alpha-arylaliphatic androgens. AB - The JAr choriocarcinoma cell cultures have demonstrated high levels of aromatase activity and have been useful for assaying a wide variety of aromatase inhibitors for aromatase inhibition in intact cells. Recently, several 7alpha-arylaliphatic androgens have shown effective inhibition of human placental microsomal aromatase in vitro, with apparent Ki values ranging from 10 to 20 nM. A series of 7alpha arylaliphatic androst-4-ene-3,17-dione compounds demonstrated potent competitive inhibition, and 7alpha-arylaliphatic androsta-1,4-diene-3,17-diones were enzyme activated irreversible inhibitors. Both series of these potent inhibitors were investigated for the ability to inhibit aromatase activity in JAr cells by measuring the conversion of [1beta-3H]-androstenedione to 3H2O and unlabelled estrone. JAr cell cultures were incubated for 2 h at 37 degrees C with the aromatase inhibitors at concentrations of 10 pM to 10 microM, the percentage of enzyme inhibition was determined, and IC50 values for inhibitors were calculated. Both series of synthetic compounds demonstrated good to excellent aromatase inhibition, and the most effective inhibitors in both series were those compounds with a phenylpropyl substituent at the 7alpha-position of the steroid nucleus. The 7alpha-arylaliphatic androst-4-ene-3,17-diones exhibited inhibition of JAr aromatase activity with IC50 values from 300 to 434 nM. More potent aromatase inhibition was observed with the 7alpha-arylaliphatic androsta-1,4-diene-3,17 diones, which exhibited IC50 values from 64 to 232 nM. Enhanced efficacy of steroidal enzyme-activated irreversible inhibitors compared to competitive inhibitors was observed in these studies and is consistent with previous reports. These results suggest that JAr choriocarcinoma cells with high levels of aromatase activity may be useful in differentiating steroidal aromatase inhibitors exhibiting different mechanisms of enzyme inhibition. In summary, the 7alpha-phenylpropyl androsta-1,4-diene-3,17-dione analogs, which are enzyme activated irreversible inhibitors, demonstrated the most effective inhibition of aromatase activity present in the JAr cell cultures among the various 7alpha arylaliphatic androgens. PMID- 9328213 TI - Up-regulation of the estrogen receptor by triiodothyronine in rat pituitary cell lines. AB - The effects of thyroid hormones on estrogen receptor (ER) levels in four different rat pituitary clonal cell lines designated as MtT/Se, SM, S and E, were investigated. When T3 was added at 10(-7) M, a significant increase in ER was evident after 9 h with maximum levels reached after 24-48 h in MtT/Se (270% of control), MtT/SM (160%) and MtT/S (140%). No significant increase was noted in MtT/E cells in which the T3 receptor (T3R) level was only one-tenth of that in the other cells, suggesting a direct link between ER expression and T3R binding. ER levels began to increase at 10(-10) M and reached maxima at 10(-8) M in MtT/Se, SM and S cells. Up to 10(-5) M of retinoic acid (RA) did not change ER levels in any of the cell lines. When Se cells were treated with cycloheximide before T3 administration, the increase in ER was completely blocked. Northern blot analysis of total RNA isolated from T3-treated MtT/Se cells revealed a limited 1.4-fold increase in ER mRNA at 10(-7) M. In conclusion, thyroid hormones (but not RA) increase ER levels in pituitary cells by a process requiring protein synthesis, and which is accompanied by an increase in the mRNA. PMID- 9328214 TI - The androgen-dependent mouse seminal vesicle secretory protein of 99 amino acids (MSVSP99): regulation of the mRNA and preliminary characterization of the promoter. AB - MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids) is a member of the rat and mouse seminal vesicle secretory protein (SVS) family. In order to characterize its androgenic regulation, the cloned cDNA and gene encoding MSVSP99 have been used. At adulthood, the MSVSP99 mRNA represents from 3 to 7% of the total mRNA population. This mRNA accumulation is under androgenic control because it is abolished by castration and restored in castrated mice by heptylate testosterone injection. During ontogenesis, MSVSP99 mRNA is just detectable in 10 day-old mice, and reaches adult levels at 30 days. Neonatal castration abolishes MSVSP99 mRNA accumulation in 20-day-old mice. Transcription elongation assays show that androgens act mainly on the MSVSP99 gene transcription. In an attempt to obtain information about the mechanism of androgen action on transcription, preliminary transient transfection experiments in CV-1 cells permitted us to define a promoter region (-387/ + 16), the activity of which is enhanced by dihydrotestosterone. PMID- 9328215 TI - Evaluation of the major metabolites of raloxifene as modulators of tissue selectivity. AB - Raloxifene (LY139481 HCl) is a selective estrogen receptor modulator (SERM) which blocks the effects of estrogen on some tissues, such as the breast and uterus, while mimicking estrogen in other tissues, such as bone. To study the origins of this unique pharmacology, we have prepared the major metabolites of raloxifene as chemical probes for examining the estrogen receptor function in vitro and in vivo. In human breast cancer cell (MCF-7) related assays, these glucuronide conjugates show little affinity for the estrogen receptor and are more than two orders of magnitude less potent at inhibiting cell proliferation than raloxifene. In non-traditional estrogen target tissue, such as bone, these metabolites are less effective than the parent at inhibiting cytokine-stimulated bone resorbing activity in rat osteoclasts or producing transforming growth factor beta-3 (TGF beta3). In animal models, tissue distribution studies with radiolabelled metabolite indicate that conversion to raloxifene occurs readily in a variety of tissues including the liver, lung, spleen, kidney, bone and uterus. Differential conversion of metabolite in target organs, such as bone and the uterus, is not observed indicating that the origin of raloxifene's pharmacology does not result from tissue-selective deconjugation of metabolite to parent. PMID- 9328216 TI - Safety and kinetic properties of a humanized antibody to human interleukin-6 receptor in healthy non-human primates. AB - A monoclonal antibody, hPM-1, was constructed by grafting the complementarity determining regions to human interleukin-6 (IL-6) receptor, raised in mouse, onto a human antibody backbone (humanized antibody). It is expected to be useful as a therapeutic agent for IL-6-related diseases such as multiple myeloma. To investigate the toxicological and kinetic properties of hPM-1 preliminarily, normal cynomolgus monkeys, which showed cross-reactivity with hPM-1, were intravenously administered with hPM-1 at doses of 0 (vehicle), 4 or 40 mg/kg once a week for 13 weeks. Upon toxicological examination, there were no changes in clinical signs, food consumption, body weights, urinalyses, body temperatures, electrocardiograms, hematological and biochemical parameters including blood platelet counts, serum levels of immunoglobulin G and C-reactive protein, and pathological findings. In a kinetic study, serum concentrations of hPM-1 showed a linearity between doses of 4 and 40 mg/kg. The serum concentrations, even at a dose of 4 mg/kg, were maintained at a high enough level to inhibit the IL-6 functions throughout the period of the study. Concentrations of hPM-1 in bone marrow were almost equal to those in serum. The antibodies against hPM-1 were detected only in one of four monkeys receiving hPM-1. This study suggests that blockage of the IL-6 receptor by hPM-1 does not induce any influence on a healthy living body, and hPM-1 is not toxic under the conditions of this investigation. PMID- 9328217 TI - Methylmercury alters the tyrosination status of tubulin in the brains of acutely intoxicated rats. AB - Tyrosination/detyrosination, a post-translational modification at the carboxyl terminus of alpha-tubulin, was investigated in the brain cytosol fraction of rats treated with methylmercury (MeHg) chloride (10 mg/kg per day, for 7 days). The amount of detyrosinated tubulin species, determined as the incorporation of 14C tyrosine at the carboxyl-terminal end of alpha-tubulin, was significantly decreased throughout the experimental period of MeHg intoxication. Furthermore, the activity of tubulin-tyrosine ligase, as well as the amounts of tyrosinatable tubulin determined and calculated by a method involving pancreatic carboxypeptidase A, also decreased in the latent and symptomatic periods. Tubulin tyrosine carboxypeptidase activity did not change during the MeHg intoxication. The total amounts of alpha- and beta-tubulins, as determined by densitometry and immunoblotting, did not show significant changes during the intoxication. These results suggest that MeHg treatment may produce perturbation of cellular activities associated with the tubulin/microtubule system by altering the tyrosination status of tubulin in the rat brain. PMID- 9328218 TI - Murine strain differences in allergic airway inflammation and immunoglobulin production by a combination of antigen and diesel exhaust particles. AB - To clarify the relationship between the manifestations of allergic airway inflammation modulated by diesel exhaust particles (DEP) and immunoglobulin production in response to an antigen, airway inflammation characterized by the infiltration of eosinophils, goblet cell proliferation, and antigen-specific immunoglobulin (Ig) production was investigated in five strains of mice after immunization with ovalbumin (OA). Mice were injected intratracheally with OA (1 microg) or OA (1 microg) + DEP (50 microg) four times at 3-week intervals. The order of antigen-specific IgG1 production in plasma of mouse strains treated with OA alone was CBA/2N lactational > placental). The DTH response to BSA was suppressed in the males receiving both placental and lactational exposure. These results suggest that the immunotoxic effects of perinatal TCDD exposure of rats persist into adulthood and that suppression of the DTH response may represent the most sensitive biomarker for TCDD-induced immunotoxicity in this species. PMID- 9328224 TI - Comparative studies of the biological distinction between unipolar and bipolar depressions. AB - Although unipolar depression and bipolar depression are considered distinct entities both by clinicians and researchers, it is not clear whether a pathophysiological distinction, which is the bridge between etiology and treatment, exists between these two conditions. The objective of this paper was to systematically review the studies that examined the biological differences between unipolar and bipolar depression. Using computerized Medline and manual searches, we located and reviewed studies that directly compared patients with unipolar depression with bipolar depressed patients on at least one biological variable. The results showed that patients with bipolar depression had lower levels of urinary NE and its metabolites and lower platelet MAO activity, and higher platelet free and stimulated intracellular calcium levels compared with unipolar depressed patients, but none of the variables examined appeared to differentiate the two groups consistently. We discuss some of the methodological flaws that might have contributed to this, and suggest that further studies should control for such confounding variables. PMID- 9328225 TI - Physiological and pharmacological properties of an endogenous sodium pump inhibitor. AB - To investigate on Na+, K+-ATPase behavior in chronic uremia, pre and postdialysis serum from 10 chronic dialysis patients and 10 healthy subjects was pooled and subjected to reverse phase C-18 HPLC. Only one fraction, isolated from pre and postdialysis sera, eluting at 28 min (F1), was found to display significant effects on electrophysiological and transepithelial 22Na flux pattern of rabbit distal colon mucosa mounted in Ussing type chambers; indeed, serosal addition of uremic F1 to colonic mucosa resulted in a slow, but constant, decline in short circuit current (Isc) (deltaIsc = 1.55+/-0.16 microEq h(-1) cm(-2), mean +/- S.E.M., n=12, p<0.01) and transepithelial conductance (G(T)) (from 4.50+/-0.23 to 3.71+/-0.33 mS cm(-2), p<0.01, n=12). Measurement of transepithelial 22Na fluxes in the presence of pre or postdialysis sera also showed a significant Na+ absorption rate decrease (from 1.3+/-0.22 to 0.48+/-0.30 microEq h(-1) cm(-2), mean +/- S.E.M., n=4, p<0.01), mainly due to a decrease in mucosal-to-serosal Na+ flux. By contrast, assays of peaks isolated from healthy sera did not inhibit Isc and transepithelial Na+ transport. The incubation of highly purified basolateral membranes with F1 for 1 min produced a approximately 26% inhibition of Na+, K+ ATPase. These findings are consistent with the presence of an endogenous inhibitor of sodium pump activity in uremic plasma; it is of pharmacological interest in that it may participate in the development of unpredictable responsiveness to digitalis therapy in pathophysiologic states. PMID- 9328226 TI - Effect of docarpamine, a novel orally active dopamine prodrug, on the formation of free and sulfoconjugated dopamine in patients who underwent cardiac surgery. AB - To evaluate the clinical efficacy of orally active dopamine prodrug, docarpamine [N-(N-acetyl-L-methionyl)-O,O-bis (ethoxycarbonyl) dopamine], we examined its effect on the formation of free and sulfoconjugated dopamine in patients who underwent cardiac surgery. The preoperative values of free and sulfoconjugated dopamine in patients were 216 +/- 52 pg/ml and 4,930 +/- 820 pg/ml, respectively. The plasma level of free dopamine increased to 95.3 +/- 28.3 ng/ml by dopamine infusion after the operation and was sustained at a high level (87.7 +/- 26.5 ng/ml) by concomitant administration of docarpamine in spite of tapering of dopamine infusion. After stopping dopamine infusion, plasma level of free dopamine was 24.5 +/- 17.6 ng/ml maintained by oral administration of docarpamine alone. From these results, docarpamine may be a useful alternative to intravenous dopamine after cardiac surgery. Sulfoconjugated dopamine in plasma increased to 267 +/- 120 ng/ml after the start of dopamine infusion and increased further after oral docarpamine administration to 2,060 +/- 610 ng/ml. Since sulfoconjugated dopamine is thought to be a possible precursor of active free dopamine in plasma, orally administered docarpamine might be stored as a reserve pool for free dopamine in patients who undergo cardiac surgery. PMID- 9328228 TI - Effect of gonadotrophin and sex steroid on the scotoresponses of day old chicks of Japanese quail, Coturnix coturnix japonica. AB - Luteinizing hormone (LH) and testosterone propionate (TP) were tested for their effects on the development of scotorefractoriness in Japanese quail. Day old chicks of both the sexes were divided into four groups and treated with normal saline, LH (1 microg/100 g body weight) and two doses of TP (100 microg/100 g body weight- TP1 and 1 mg/100 gm body weight- TP2) over a periods of 14 weeks. In the male chicks, compared to control, LH treatment advanced and low dose of TP suppressed the development of scotorefractoriness, while high dose of TP inhibited it completely and maintained the birds in scotosensitive state. On the other hand, in females, LH treatment increased the rate of sexual development resulting in the onset of egg laying earlier than that of control, but both the doses of steroid hormone suppressed ovarian development. It is suggested that, LH treatment not only induced a higher degree of reproductive development in short day quail but may also advance sexual maturity as under long daylength. Further, both the doses of male hormone had negative feedback effect on neuroendocrine axis and eliminated the attainment of scotorefractoriness i.e., reproductive development under short days. PMID- 9328227 TI - Direct inhibitory effect of adrenomedullin and guanylin on isolated caecal circular smooth muscle cells of guinea pig. AB - Smooth muscle cells isolated from the caecal circular smooth muscle layers of the guinea pig were used to determine whether adrenomedullin and guanylin can inhibit the contractile response produced by 10(-9) M cholecystokinin octapeptide (CCK 8). In addition, to elucidate each intracellular mechanisms, we examined the effects of an inhibitor of cAMP-dependent protein kinase, an inhibitor of particulate guanylate cyclase, and an inhibitor of soluble guanylate cyclase on the adrenomedullin- or guanylin-induced relaxation of the caecal circular smooth muscle cells. Both adrenomedullin and guanylin inhibited the contractile response produced by CCK-8 in a dose-dependent manner, with IC50 values of 0.12 nM and 2.4 pM, respectively. An inhibitor of cAMP-dependent protein kinase significantly inhibited the relaxation produced by adrenomedullin. In contrast, an inhibitor of particulate guanylate cyclase and an inhibitor of soluble guanylate cyclase did not have any significant effect on the relaxation produced by adrenomedullin. On the other hand, an inhibitor of particulate guanylate cyclase significantly inhibited the guanylin-induced relaxation, although an inhibitor of cAMP dependent protein kinase and an inhibitor of soluble guanylate cyclase did not have any significant effect on the guanylin-induced relaxation. In this study, we first demonstrated the direct inhibitory effects of adrenomedullin via cAMP system and guanylin via particulate guanylate cyclase system on the isolated caecal circular smooth muscle cells. PMID- 9328229 TI - Molecular cloning, characterization and cellular distribution of rat steroidogenic acute regulatory protein (StAR) in the ovary. AB - Rat ovarian genes induced by the treatment of immature rats with pregnant mare serum gonadotropin (PMSG) were isolated by a subtraction cloning method. Amongst them was obtained a probable rat homologue of steroidogenic acute regulatory protein (StAR), which has been recently identified as a protein that is an acute regulator of the rate limiting transfer of cholesterol from the outer to the inner mitochondrial membrane. Structure of rat StAR was determined by nucleotide sequence analysis. Northern blot analysis revealed that StAR mRNA levels were rapidly and strongly increased by PMSG/hCG but not by FSH. In situ hybridization revealed that the expression of StAR mRNA was strongly induced by PMSG in theca interna cells as well as in corpora lutea. These findings indicate that expression of StAR mRNA is restricted to and induced in the ovarian steroidogenic cell types where cholesterol is used as a substrate for synthesis of steroid hormones. PMID- 9328230 TI - Ovarian nitric oxide synthase (NOS) gene expression during peripubertal development. AB - Nitric oxide (NO) is generated from L-arginine by different isoforms of the enzyme nitric oxide synthase (NOS) and is known to be involved in mediating several biological functions, some of which are associated with reproduction. In this study, we examined the ability of the prepubertal ovary to express the inducible (i), as well as the neuronal-type constitutive (c) form of NOS and also, investigated whether either isoform undergoes changes in the ovary during peripubertal development. Results indicate that both forms of NOS were expressed in the ovary and that the iNOS mRNA transcripts were expressed without being exogenously induced. When compared with juvenile levels, iNOS, but not cNOS, mRNA increased (p<0.01) during the early proestrous phase of development. By the late proestrous phase, the levels of iNOS mRNA declined markedly (p<0.001) and remained low throughout both the first estrous and diestrous phases. Western blot analysis revealed both iNOS and cNOS protein expression in each phase of puberty with only iNOS showing a significant change during the peripubertal period. Specifically, there was an initial increase in the expression of iNOS protein during the late proestrous phase (p<0.05) which was accompanied by preovulatory increases in serum estradiol (p<0.01) and LH (p<0.001). The iNOS protein levels then dramatically increased to peak on the morning of first estrus (p<0.001), an event associated with declining (p<0.05) serum levels of estradiol. These data demonstrate developmental changes in the expression of ovarian iNOS mRNA and protein both before and after first ovulation; hence, suggesting a role for NO in the ovary during pubertal maturation and furthermore, providing compelling evidence at the gene level supporting the hypothesis that the NO/NOS system plays a physiological role in ovarian function. PMID- 9328231 TI - A single dose of mifepristone induces ovulation in pseudopregnant rats. AB - We used mifepristone (M) to evaluate the role of progesterone in maintaining pseudopregnancy. Cycling rats were made pseudopregnant (psp) by cervico-vaginal stimulation (CVS) on the day of estrus (day 0) and received 10 mg/kg M or vehicle (control groups) on day 3. Blood samples were taken at 06.00 h on days 4, 6 or 7 or at 18.00 h on days 3, 4, 6 or 10. M induced proestrus 2 days later (day 5), estrus on day 6, and a second prolonged diestrus afterwards. Prolactin and progesterone levels were similar in the control and M treated groups excepting on day 6, when both were reduced in the M-treated animals, and these rats were in estrus, suggesting a temporary impairment of luteal function. To demonstrate activated corpora lutea the endometrium was scratched on the fourth day of the first or second diestrus in additional control and M-treated groups. The deciduomal response was seen in the control and M groups after scratching the endometrium on day 4 of the first or second diestrus, respectively, but M blocked the deciduomal response in the first diestrus. Ovulation was confirmed by finding that 66.7% of the M-treated rats showed ova in the Fallopian tubes on the M induced estrus and 4 out of 10 of the M rats placed with males on the M-induced proestrus showed spermatozoa in the vaginal smears. Half of these became pregnant, delivering 2 pups each. The results show that M can induce ovulation in psp rats, demonstrating that the anovulation observed after CVS is dependent on progesterone, yet luteal function persists after M in pseudopregnancy. Progesterone may act either by suppressing LH secretion or by permitting prolactin secretion, or both. Moreover, progesterone is required to maintain endometrial responsiveness. PMID- 9328232 TI - Non-epithelial endothelin-A receptors activate adenylate cyclase in rat trachea: biochemical mechanisms and physiological implications. AB - In the present study, we investigated the mechanisms underlying the differential effects of endothelins (ETs) in the rat trachea. Sarafotoxin-S6c (SRTX-c) and ET 3 were more potent spasmogens to rat tracheal strips than ET-1. The EC50 values were 12, 14.1 and 89.1 nM, respectively. Tension responses to ET-1 and ET-3, but not to SRTX-c, were enhanced by either indomethacin or the ET(A) blocker, BQ-610 (1 microM). In epithelium-intact tracheal rings, both ET-1 and ET-3 activated adenylate cyclase (AC) in a concentration-dependent manner. The activation by ET 1 of AC was significantly higher than that of ET-3. Thus, EC50 values for ET-1 and ET-3 were 71 and 200 nM, and maximal cAMP increments were 196% and 62% above baseline, respectively. SRTX-c, up to 1 microM, did not alter basal cAMP level. Mechanical removal of the epithelium neither had an effect on AC activation by ET 1 or ET-3, nor did it alter the inability of SRTX-c to modulate AC activity. Conversely, pre-incubation of tracheal strips with indomethacin (1 microM) virtually ablated the increments in cAMP by the ETs. Likewise, BQ-610 attenuated AC activation, concentration-dependently (IC50=28.2 nM). Taken together, the present study suggests that ET(A) receptors, from non-epithelial source, are functionally-linked to AC activation via a prostanoid-dependent pathway. This ET(A)-initiated cascade acts to negatively regulate muscle contraction. Such a cross-talk between ET signals most likely accounts for variation of tension responses to ET homologs. PMID- 9328234 TI - Ochre suppressor transfer RNA restored dystrophin expression in mdx mice. AB - The mdx mouse is an animal model for human Duchenne muscular dystrophy. The lack of dystrophin in mdx mice is caused by an ochre mutation in exon 23 of the dystrophin gene. This study tested the feasibility of inhibiting translational termination as an approach for genetic therapy for diseases caused by nonsense mutations. We evaluated both the in vitro and in vivo efficiencies of readthrough of ochre codons in 2 genes with the tRNA suppressor gene. The first target was a CAT reporter gene bearing an ochre mutation at the 5' end (CATochre). The second target was the dystrophin gene in mdx mice. The readthrough efficiencies were about 20% in COS cells and 5.5% in rat hearts. At four weeks after a direct injection of plasmid DNA encoding the tRNA suppressor into mdx mice, dystrophin positive fibers were detected by sarcolemmal immunostaining. This is the first convincing data that a tRNA suppressor gene might be a useful in vivo treatment for the genetic disorders caused by nonsense mutations. PMID- 9328233 TI - The marine toxin okadaic acid reduces O2- generation and tyrosine phosphorylation in LPS-primed rat neutrophils. AB - Contrasting effects of okadaic acid (OKA) on neutrophil (PMN) superoxide anion (O2-) generation have been reported. In this study, we examined the effect of OKA on phorbol myristate acetate (PMA)-stimulated O2- generation in rat PMNs primed with LPS in vivo (LPS-PMN) and saline-treated rat PMNs (SAL-PMN). The following results were observed: (1) OKA, but neither genistein nor vanadate, markedly reduced O2- generation in a dose and time-dependent manner; (2) genistein, a tyrosine kinase inhibitor, as well as OKA, reduced tyrosine phosphorylation; (3) sodium orthovanadate, a tyrosine phosphatase inhibitor, potently enhanced tyrosine phosphorylation. Our studies suggest that OKA might reduce tyrosine phosphorylation by affecting the activity of tyrosine phosphatases regulated by serine-threonine phosphorylation. PMID- 9328235 TI - Effects of dietary Spirulina maxima on endothelium dependent vasomotor responses of rat aortic rings. AB - The aim of this study was to evaluate the effects of Spirulina maxima on vasomotor responses of aorta rings from male Wistar rats fed on a purified diet. For this purpose, the animals (weighing 200-240 g) were allocated randomly in two groups. One receiving purified control diet (A) and the other receiving purified diet containing 5% Spirulina (B). Purified diets were according to American Institute of Nutrition guidelines and adjusted to Spirulina protein content. All animals were fed (20 g/day/rat) during two weeks, receiving water ad libitum and 12 h. light-dark cycles. Spirulina maxima effects were evaluated by concentration response (CR) curves of aorta rings with or without endothelium to phenylephrine (PE), both in presence and absence of indomethacin (Indom) or indomethacin plus L NAME (Indom. + L-NAME), and to carbachol (CCh). Aorta rings with endothelium from group B showed, relative to corresponding rings from group A: 1) a significant decrease in the maximal tension developed in response to PE. 2) this decrease was reverted by Indom. 3) Indom. + L-NAME induced an additional increase in the contractile responses to PE. 4) a significant shift to the left of the CR curve to CCh. No significant differences were observed in the tension developed in response to PE in rings without endothelium from either group. These results suggest that Spirulina maxima may decrease vascular tone by increasing the synthesis and release of both a vasodilating cyclooxygenase-dependent product of arachidonic acid and nitric oxide, as well as by decreasing the synthesis and release of a vasoconstricting eicosanoid from the endothelial cells. PMID- 9328236 TI - GABA(A) antagonist and nicotine-induced antinociception. AB - Rats, pretreated with saline or GABA(A) antagonist bicuculline (BIC) at the doses of 2, 4, 6, 10 and 20 mg/kg, were injected with nicotine (NIC, 1 mg/kg) 30 min later. Tail-flick (TF) latencies were measured before (baseline) and three times at 10 min interval after pretreated compounds, continued every 10 min up to 1 hr after NIC injection. In all groups, median TF latencies did not change from the baseline for the first 30 min after the pretreatment. Following NIC alone (control) and in the group pretreated with 2 mg/kg BIC, 60% rats reached ceiling TF latencies (20 sec) lasting for 10 min. In groups with higher BIC doses (4 to 10 mg/kg), median TF latencies were in the range of 5-7 sec with 30% rats reaching the ceiling TF latencies. Following 20 mg/kg BIC, one out of five rats reached 20 sec; the median was in the range of 4-5 sec. Significantly lower responses were observed following 4 mg BIC and higher doses with no difference among the groups. In conclusion, our novel data show that BIC alone, injected systemically, does not possess any effect on the thermal nociceptive transmission as measured by the tail flick test. However, pretreatment with BIC partially prevents NIC-induced antinociception, in a non dose related manner. This suggests that GABA(A) receptors may, at least in part, contribute to the complex mechanisms involved in NIC-induced antinociception. PMID- 9328237 TI - Services for patients with multiple sclerosis. PMID- 9328239 TI - Allesandro Volta (1745-87). PMID- 9328238 TI - Neurology and the liver. PMID- 9328240 TI - Neurological picture. Coma nails. PMID- 9328241 TI - Neurological picture. A "giant" prolactinoma. PMID- 9328242 TI - Disturbed striatoprefrontal mediated visual behaviour in moderate to severe parkinsonian patients. AB - OBJECTIVES: To study visuospatial and visual memory functions in moderate to severe parkinsonian patients. METHODS: Visual antisaccades (AS) and remembered saccades (RS) were examined in 30 patients with moderate to severe Parkinson's disease and in 44 age matched controls. AS are saccades in the direction opposite to the target. RS are saccades towards the remembered position of a target that is no longer visible. RESULTS: Patients with Parkinson's disease had serious difficulties in suppressing a reflex saccade ("visual grasping") or they made no saccade at all ("visual akinesia"). The remembered saccade was often wrongly directed. CONCLUSIONS: These types of errors point to a dysfunction in the striatoprefrontal loop. The discrepancy of the results with those in the literature, which are mostly normal, could be explained by the more advanced stage of our patients. This might correspond to the development of unresponsiveness to levodopa or of non-dopaminergic lesions. PMID- 9328243 TI - Putaminal petechial haemorrhage as the cause of chorea: a neuroimaging study. AB - OBJECTIVES: A hyperintense putamen on either CT or MRI as a finding associated with chorea has occasionally been described and is almost always associated with non-ketotic hyperglycaemia. The cause of the hyperintensity of the striatum in these images is still controversial. Some reports have found that calcification was responsible whereas others have advocated petechial haemorrhage as the cause. The purpose of this study was to determine whether hyperintense striata are caused by petechial haemorrhage or calcification, with the sequential imaging changes. SUBJECTS AND METHODS: Five patients presenting with an acute onset of either hemichorea or generalised chorea and showed either unilateral or bilateral hyperdense striatum on the initial CT were assessed. Neuroimaging studies including sequential CT and MRI examinations and detailed biochemical tests were performed. RESULTS: Three patients had pronounced hyperglycaemia and the other two patients had no biochemical abnormalities. In all patients, the first CT scans, performed within two weeks of the onset of chorea, showed a high density over the striatum contralateral to the chorea, which diminished or disappeared two months later. T1 weighted imaging disclosed hypersignal intensities over the striatum contralateral to the chorea on admission which diminished two months later. T2 weighted imaging at two months showed hyposignal intensity changes corresponding to the area with hypersignal changes on T1 weighted images, implying haemosiderin deposition. CONCLUSION: Based on the evolution of clinical manifestations and the findings of neuroimaging, putaminal petechial haemorrhage might be a new entity causing either hemichorea or generalised chorea. PMID- 9328244 TI - GTP cyclohydrolase I mutations in patients with dystonia responsive to anticholinergic drugs. AB - OBJECTIVES: To investigate the hypothesis that GTP cyclohydrolase I (GCH1) mutations are responsible for the phenotype of highly anticholinergic responsive dystonia in patients with apparent primary torsion dystonia. METHODS: From 107 British patients with clinically diagnosed primary torsion dystonia, seven patients were identified with an excellent response to anticholinergic drugs. All six exons of the GCH1 gene were sequenced in these patients to identify mutations. RESULTS: Three novel GCH1 mutations were identified in two patients. One patient was a compound heterozygote with asymptomatic carrier parents. The clinical phenotype of patients with and without GCH1 mutations was similar. CONCLUSIONS: These findings show that a proportion of patients with apparent primary torsion dystonia and a good response to anticholinergic drugs have GCH1 mutations and therefore have a variant of dopa responsive dystonia. The difficulty in distinguishing clinically between patients with and without mutations underscores the importance of considering the diagnosis of a levodopa responsive dystonia in all such patients. PMID- 9328245 TI - Binswanger's "encephalitis subcorticalis chronica progressiva". PMID- 9328246 TI - Deficits on self ordered tasks associated with hyperostosis frontalis interna. AB - A 74 year old patient, EW, with dorsolateral frontal cortical compression due to hyperostosis frontalis interna, in the absence of the Morgagni or Stewart-Morel syndromes, is described. In addition to conventional neuropsychological measures EW was administered one nonspatial and two spatial self ordered working memory tasks, as well as a standard measure of fluid intelligence or g. She showed impaired performance on all three self ordered working memory tasks compared with a normal control group of 10 subjects matched for age, education, sex, and IQ. By contrast, her performance on the fluid intelligence test was comparable with that of the controls. It is concluded that the compression of dorsolateral frontal cortex accompanying hyperostosis frontalis interna may produce selective cognitive impairment. PMID- 9328247 TI - The nucleus basalis (Ch4) in the alcoholic Wernicke-Korsakoff syndrome: reduced cell number in both amnesic and non-amnesic patients. AB - BACKGROUND: The cholinergic nucleus basalis (Ch4) is an exclusive site of neurofibrillary degeneration in alcoholic patients with Wernicke's encephalopathy. AIM: To test the hypothesis that the loss of Ch4 neurons contributes to the memory disorder, Korsakoff's psychosis, commonly seen in Wernicke's encephalopathy. METHODS: Magnocellular basal forebrain neurons were quantified in alcoholic patients with Wernicke's encephalopathy, both with and without Korsakoff's psychosis, and neurologically asymptomatic alcoholic and non alcoholic controls. Because amnesic and non-amnesic patients with Wernicke's encephalopathy share common periventricular lesions, both thiamine deficient groups as well as alcoholic patients with no neurological complications were included to determine the lesion specific to memory impairment. RESULTS: Ch4 cell number did not differ significantly between alcoholic and non-alcoholic controls and there was no correlation between cell number and lifetime alcohol intake. However, Ch4 cell number in all groups was significantly correlated with the volume of its major projection target, the cerebral cortex. Ch4 cell number in the non-amnesic Wernicke's encephalopathy group was significantly below controls (24%), with cell number in patients with Korsakoff's psychosis 21% below controls. There was considerable overlap in cell number between groups. On discriminant analysis, there was significantly greater cell loss in three non amnesic patients with Wernicke's encephalopathy than in some patients with Korsakoff's psychosis. The nonamnesic patient with the greatest cell loss was impaired on attentional tasks. CONCLUSION: Whereas neurons in the nucleus basalis are at risk in thiamine deficient alcoholic patients, cell loss is minor and does not account for the profound memory disorder. PMID- 9328248 TI - Herpes simplex encephalitis treated with acyclovir: diagnosis and long term outcome. AB - OBJECTIVES: The frequency and characteristics of the long term sequelae of herpes simplex encephalitis were assessed after treatment with acyclovir. METHODS: Patients were included if they were treated with acyclovir and the diagnosis of herpes simplex encephalitis was confirmed by culture of herpes simplex virus (HSV) from the brain, an increase in the CSF HSV antibody titre, or detection of HSV deoxyribonucleic acid in the CSF. Each patient's medical records were reviewed and surviving patients were interviewed and examined. RESULTS: A diagnosis of herpes simplex encephalitis was confirmed in 42 patients. Five patients (12%) died in the first month. Three patients (7%) had severe neurological sequelae and died after a longer interval. All but one of the 34 surviving patients had neurological symptoms, an abnormal neurological examination, or both. Twenty patients (48%) performed everyday activities as well as before herpes simplex encephalitis; nine patients (21%) were living independently, but were functioning at a lower level than before the illness; and five patients (12%) had a severe neurological deficit. Twenty nine of the 34 survivors were assessed six months to 11 years after herpes simplex encephalitis. The most common long term symptoms were memory impairment (69%), personality and behavioural abnormalities (45%), and epilepsy (24%). Short term memory impairment (70%), anosmia (65%), and dysphasia (41%) were the most common signs. CONCLUSIONS: Although acyclovir has reduced the mortality of herpes simplex encephalitis, 30% of this group of patients either died or had a severe neurological deficit. The other 70% of the patients regained independence in activities of daily living, but most of these people had persistent neurological symptoms, signs, or both. PMID- 9328249 TI - Non-invasive investigations successfully select patients for temporal lobe surgery. AB - OBJECTIVES: There is controversy regarding the need for invasive monitoring in the preoperative assessment of patients with temporal lobe epilepsy. The use of a series of non-invasive investigations in identifying the seizure focus is reported in 75 consecutive adults referred for epilepsy surgery. METHODS: All had video-EEG monitoring using scalp electrodes, high resolution MRI, and neuropsychology assessment. Other investigations included volumetric MRI, PET, and ictal and interictal SPECT. The seizure focus was localised and surgery offered if MRI disclosed unilateral hippocampal atrophy or a foreign tissue lesion and other investigations were either concordant or not discordant. RESULTS: In 68 patients the seizure focus was localised and three patients were inoperable. Sixty five patients have been offered surgery and 50 have undergone temporal lobe surgery and have a follow up of at least 12 months (mean 24 months). All had pathology: hippocampal sclerosis 34, dysembryoblastic neuroepithelial tumour six, cavernoma four, dysplasia two, low grade glioma two, ganglioglioma two. Thirty nine patients (78%) are seizure free postoperatively, 29/34 with hippocampal sclerosis and 10/16 with a foreign tissue lesion. Of the 11 patients with postoperative recurrent seizures, eight have a >90% reduction in seizure frequency and three have <90% reduction in seizure frequency but a worthwhile improvement. CONCLUSIONS: Non-invasive investigations successfully select most patients for temporal lobe surgery. PMID- 9328250 TI - Haemostatic changes during surgery for primary brain tumours. AB - OBJECTIVE: Primary brain tumours may be associated with coagulation disorders which can pose intraoperative and postoperative management difficulties. The aim was to evaluate the coagulation profile of patients with brain tumours undergoing surgery using thromboelastography (TEG) in combination with simple laboratory tests. METHODS: Fifty adult patients with primary brain tumours larger than 4 cm in maximum diameter and no history of coagulation disorders were studied in a prospective, observational manner over a one year period. Preoperative, intraoperative, and postoperative measurements included haemoglobin concentration, platelet count, prothrombin and partial thromboplastin times, fibrin(ogen) degradation product concentration, D-dimer concentration, and TEG. RESULTS: Eleven patients (22%) had abnormal intraoperative TEGs, of whom six (12%) subsequently developed haematomas requiring surgical evacuation. The coagulopathy seemed to be hyperfibrinolysis in two cases (4%) and disseminated intravascular coagulation in four (8%). There was no preoperative difference in reaction time (R time) for clot formation between the non-haematoma and haematoma groups(mean 11.44 (SD 3.42) v 12.33 (2.50) min, P=0.46). However, when other preoperative indices were compared, in the non-haematoma group, K time (time to reach a clot amplitude of 20 mm) was shorter (6.72 (2.15) v 10.56 (3.50) min, P=0.001), rate of clot growth (a) was faster (43.67 degrees (7.53) v 27.11 degrees (5.42), P<0.0001) and maximum amplitude of clot strength (MA) was greater (52.64 (7.85) v 40.33 (6.59) mm, P<0.001). Intraoperatively, R time was significantly shortened in the non-haematoma group, (7.67 (1.78) min, P<0.0001) unlike the haematoma group (10.67 (1.58) minutes, P=0.11). CONCLUSIONS: Although these results indicate a general hypercoagulability during brain tumour surgery, in certain cases, a predisposition towards hypocoagulability may exist even before surgery, detectable only when the physical characteristics of clot formation are studied by TEG. Judicious replacement of clotting factors, platelets, and antifibrinolytic agents should be considered intraoperatively if the TEG is abnormal, without waiting for laboratory test results. PMID- 9328251 TI - Interferon-gamma induced increases in intracellular calcium in T lymphocytes from patients with multiple sclerosis precede clinical exacerbations and detection of active lesions on MRI. AB - BACKGROUND: Interferon (IFN)-gamma exerts a multiplicity of actions potentially relevant for the pathogenesis of multiple sclerosis, including the expression of a transplasmalemma calcium (Ca2+) influx leading to an intracellular Ca2+ ([Ca2+]i) increase able to lower T lymphocyte threshold of excitability. It has been previously shown in a cross sectional cumulative study that this influx is associated with clinical and MRI evidence of disease activity. METHODS: To evaluate the temporal relation between disease activity and the IFN-gamma activated Ca2+ influx in individual patients, a fluorimetric analysis was performed on peripheral blood lymphocytes from eight patients with relapsing remitting multiple sclerosis every 15 days for one year. RESULTS: Fluctuations of the influx were correlated with clinical events and monthly enhanced brain MRI. The influx was detected a mean of 10.4 (range 7-17) times per patient during our analysis. In 61% of the occasions, influx induced [Ca2+]i increases were recorded in each patient in more than two consecutive measurements, determining sustained [Ca2+]i increases lasting for a mean of 31.5 days. Peak [Ca2+]i increases preceded clinical attacks (P=0.04) or maximal detection of brain MRI enhancing lesions (P=0.05) by a mean of 30.8 and 34.2 days respectively. Spectral analysis of time series further indicated that the fluctuation frequency of [Ca2+]i increases due to the influx over time were superimposable on the appearance of new MRI lesions in all patients and confirmed that in two thirds of the patients these [Ca2+]i increases occurred significantly before (P<0.005) or concurred with new lesion appearance. Finally, the overall presence of the influx throughout the follow up period correlated (P=0.03) with the patients' exacerbation rates. CONCLUSIONS: Intracellular events leading to T lymphocyte activation in multiple sclerosis occur in the peripheral blood before CNS specific events become evident and are, in part, sustained by cytokine induced Ca2+ mediated phenomena. PMID- 9328252 TI - Delayed recovery of nerve conduction and vibratory sensibility after ischaemic block in patients with diabetes mellitus. AB - OBJECTIVES: To determine if the recovery of nerve function after ischaemic block is impaired in patients with diabetes mellitus relative to healthy controls. METHODS: Median nerve impulse conduction and vibratory thresholds in the same innervation territory were studied in patients with diabetes mellitus (n = 16) and age matched controls (n = 10) during and after 30 minutes of cuffing of the forearm. RESULTS: Cuffing caused a 50% reduction of the compound nerve action potential (CNAP) after 21.9 (SEM 1.6) minutes in patients with diabetes mellitus and after 10.6 (0.7) minutes in controls. After release of the cuff the half life for CNAP recovery was 5.13 (0.45) minutes in patients with diabetes mellitus and <1 minute in controls. At seven minutes after release of the cuff CNAP was fully restored in the controls whereas in patients with diabetes mellitus CNAP had only reached 75.1 (4.1)% of its original amplitude. After onset of ischaemia it took 14.6 (1.9) minutes in patients with diabetes mellitus before the vibratory threshold was doubled, whereas this took 5.8 (0.8) minutes in controls. After release of the cuff half time for recovery of vibratory threshold was 8.8 (1.0) minutes in patients with diabetes mellitus and 2.6 (0.3) minutes in controls. Ten minutes after the cuff was released the threshold was still raised (2.0 (0.3) fold) in the diabetes mellitus group, whereas it was normalised in controls. Among patients with diabetes mellitus the impaired recovery correlated with older age, higher HbA1c, and signs of neuropathy, but not with blood glucose. CONCLUSION: After ischaemia there is a delayed recovery of nerve conduction and the vibratory sensibility in patients with diabetes mellitus. Impaired recovery after ischaemic insults may contribute to the high frequency of entrapment neuropathy in patients with diabetes mellitus. PMID- 9328253 TI - Dural puncture and activated protein C resistance: risk factors for cerebral venous sinus thrombosis. AB - OBJECTIVES: Dural puncture is regarded a safe procedure when contraindications are carefully excluded and has so far not been recognised as a risk factor for cerebral venous sinus thrombosis (CVST). Five patients are described with CVST after dural puncture in the presence of additional risk factors. METHODS: In four out of five patients complete investigations for thrombophilia were performed at least one month after withdrawal of oral anticoagulation. RESULTS: In three out of four patients tested, activated protein C (APC) resistance due to heterozygous coagulation factor V R506Q mutation (factor V Leiden) was found. One patient was using oral contraceptives as a circumstantial risk factor and three had had spinal anaesthesia for surgical procedures. Family history of venous thromboembolism was negative in all patients. Retrospective evaluation of 66 patients with CVST disclosed that dural puncture was the fourth most common risk factor (8%) possibly contributing to thrombosis. CONCLUSION: Dural puncture may constitute an additional risk factor for CVST especially in patients with APC resistance or surgery. In such patients a thrombophilia screen is indicated. PMID- 9328255 TI - PET and SPECT in whiplash syndrome: a new approach to a forgotten brain? AB - Whiplash associated disorders are a medicolegally controversial condition becoming increasingly worrisome in the western world. This study was designed to evaluate perfusion and glucose metabolism in whiplash brain. Using Tc-99m bicisate (ECD) single photon emission computed tomography (SPECT) and F-18 fluorodeoxyglucose (FDG) PET, six clinically and neuropsychologically controlled patients (patient group) with whiplash syndrome and 12 normal controls (control group) were investigated. Standardised elliptical regions of interest (ROIs) were determined in three adjacent transaxial slices in the frontal, parietal, temporal, and parieto-occipital cortex, cerebellum, brain stem, basal ganglia, and thalamus. For PET, the glucose metabolic index (GMI; =ROI uptake/global uptake at the level of the basal ganglia) and, for SPECT, the perfusion index (PI; =ROI/global) were calculated. In the patient group there was significant hypometabolism and hypoperfusion in the parieto-occipital regions (on the right (R) and left (L) side) compared with the control group: PET data: GMI parieto occipital R: control 1.066 (0.081) (mean (SD)), patient 0.946 (0.065); P=0.0092, Mann Whitney. GMI parieto-occipital L: control 1.034 (0.051), patient 0.922 (0.073); p=0.0067. SPECT data: PI parieto-occipital R: control 1.262 (0.066), patient 1.102 (0.063); P=0.0039. PI parieto-occipital L: control 1.226 (0.095), patient 1.098 (0.075); P=0.0273. In some patients there was hypometabolism (>2 SD of control) in regions other than the parieto-occipital region. It is hypothesised that parieto-occipital hypometabolism may be caused by activation ofnociceptive afferent nerves from the upper cervical spine. PMID- 9328254 TI - Transient global amnesia: implicit/explicit memory dissociation and PET assessment of brain perfusion and oxygen metabolism in the acute stage. AB - OBJECTIVES: To assess explicit memory and two components of implicit memory--that is, perceptual-verbal skill learning and lexical-semantic priming effects--as well as resting cerebral blood flow (CBF) and oxygen metabolism (CMRO2) during the acute phase of transient global amnesia. METHODS: In a 59 year old woman, whose amnestic episode fulfilled all current criteria for transient global amnesia, a neuropsychological protocol was administered, including word learning, story recall, categorical fluency, mirror reading, and word stem completion tasks. PET was performed using the (15)O steady state inhalation method, while the patient still exhibited severe anterograde amnesia and was interleaved with the cognitive tests. RESULTS: There was a clear cut dissociation between impaired long term episodic memory and preserved implicit memory for its two components. Categorical fluency was significantly altered, suggesting word retrieval strategy -rather than semantic memory--impairment. The PET study disclosed a reduced CMRO2 with relatively or fully preserved CBF in the left prefrontotemporal cortex and lentiform nucleus, and the reverse pattern over the left occipital cortex. CONCLUSIONS: The PET alterations with patchy CBF-CMRO2 uncoupling would be compatible with a migraine-like phenomenon and indicate that the isolated assessment of perfusion in transient global amnesia may be misleading. The pattern of metabolic depression, with sparing of the hippocampal area, is one among the distinct patterns of brain dysfunction that underlie the (apparently) uniform clinical presentation of transient global amnesia. The finding of a left prefrontal hypometabolism in the face of impaired episodic memory and altered verbal fluency would fit present day concepts from PET activation studies about the role of this area in episodic and semantic memory encoding/retrieval. Likewise, the changes affecting the lenticular nucleus but sparing the caudate would be consistent with the normal performance in perceptual-verbal skill learning. Finally, unaltered lexical-semantic priming effects, despite left temporal cortex hypometabolism, suggest that these processes are subserved by a more distributed neocortical network. PMID- 9328256 TI - Schizencephaly associated with psychosis. AB - Schizencephaly is a rare disorder of brain development resulting in the formation of abnormal unilateral or bilateral clefts in the cerebral hemispheres. It is often accompanied by partial seizures, mental retardation, and hemiparesis. Two patients are described with clear psychotic symptoms with either unilateral or bilateral schizencephaly. The implications of the association between schizencephaly and psychosis in these patients for understanding the biology of the psychoses are discussed. PMID- 9328257 TI - Sjogren's syndrome in patients with chronic idiopathic axonal polyneuropathy. AB - OBJECTIVE: To assess the presence of symptoms and signs of Sjogren's syndrome in patients with otherwise idiopathic axonal polyneuropathy and to develop guidelines for the diagnostic approach with respect to Sjogren's syndrome in these patients. METHODS: Sixty five patients with axonal polyneuropathy in whom an aetiological diagnosis could not be made underwent (1) a standard interview focusing on ocular and oral sicca symptoms, (2) physical examination, (3) tests for objective assessment of keratoconjunctivitis sicca, (4) extensive serological investigations, and (5) a sublabial salivary gland biopsy. RESULTS: In forty nine patients a sublabial salivary gland (SSG) biopsy was performed, thereby completing the whole investigation for Sjogren's syndrome. Three of these 49 patients (all women) had an SSG biopsy specimen suggestive of Sjogren's syndrome, which, in combination with other symptoms and signs, led to a diagnosis of primary Sjogren's syndrome. CONCLUSIONS: None of the three patients with primary Sjogren's syndrome had spontaneously complained about sicca symptoms and the clinical neurological picture of them did not differ from the other patients in the study. Therefore, in patients with chronic idiopathic axonal polyneuropathy, especially in women, a systematic investigation for Sjogren's syndrome should be done, because the presence of Sjogren's syndrome may have implications for treatment and justifies a clinical follow up on a regular base. PMID- 9328259 TI - Quality of life after perimesencephalic haemorrhage. AB - Quality of life was measured by means of the sickness impact profile (SIP) questionnaire in a prospectively collected, consecutive series of 25 patients with perimesencephalic haemorrhage. A mean of two years and four months (range six months to six years) after the perimesencephalic haemorrhage, quality of life scores of the (former) patients were comparable with those of a random sample from the Dutch population. For physical aspects the patients showed even less dysfunction than controls. It is concluded that a perimesencephalic haemorrhage does not reduce quality of life or capacity to work. PMID- 9328258 TI - Missense mutation (R15W) of the connexin32 gene in a family with X chromosomal Charcot-Marie-Tooth neuropathy with only female family members affected. AB - A small family with sensorimotor neuropathy of dominant inheritance was examined. All three affected members were female. They had unusually severe symptoms and pronounced reduction of motor nerve conduction velocities with absent sensory nerve action potentials. Molecular genetic analysis disclosed a missense mutation in the connexin32 gene in codon 15 (Arg15Trp) which predicts the replacement of a basic amino acid to a non-polar amino acid in the first cytoplasmic loop of the protein. This report illustrates that in small pedigrees in which only women are affected, and which show a severe clinical phenotype, X chromosomal Charcot-Marie Tooth neuropathy should be considered as differential diagnosis. PMID- 9328260 TI - Long term melphalan-prednisolone chemotherapy for POEMS syndrome. AB - The effects of long term melphalan-prednisolone (MP) therapy was studied on 12 patients with POEMS (polyneuropathy, organomegaly, endocrinopathy, M protein, and skin changes) syndrome. Six were treated with MP every six weeks for 16 to 52 months; three also with cyclophosphamide, and three with localised irradiation for osteoclastic lesions. Five of the six survived during the follow up period and showed various degrees of lessening of their neuropathy and other symptoms. There were no serious side effects. The other six patients received treatments that included corticosteroids, short term chemotherapy, or irradiation, but not long term chemotherapy. Five showed transient lessening of their non-neurological symptoms, and one, obvious neurological improvement. Five of these six patients died from nine to 70 months after POEMS onset. The findings suggest that long term MP therapy may be an effective treatment for the POEMS syndrome. PMID- 9328261 TI - Transmission of Creutzfeldt-Jakob disease via a corneal transplant. AB - A 45 year old woman is reported who initially presented with a cerebellar syndrome, severe ataxia, and dysarthria. She rapidly deteriorated to coma vigile with bilateral myoclonic jerks, flexion rigidity, and immobility necessitating complete nursing. Her EEG showed generalised slow activity and periodic biphasic and triphasic waves. The CSF concentration of neuron specific enolase was very high. Consequently the diagnosis of Creutzfeldt-Jakob disease was established. Eight months later she died of respiratory complications. Thirty years earlier the patient had undergone corneal transplantation for keratoconus. Review of the organ donor's hospital records showed that death was caused by intercurrent pneumonia subsequent to subacute spongiform encephalopathy confirmed by necropsy. In view of two previous case reports in the literature it is presumed that the cadaveric cornea was the source of transmission of Creutzfeldt-Jakob disease in this patient. PMID- 9328262 TI - Central facial weakness due to medial medullary infarction: the course of facial corticobulbar fibres. AB - Two patients are reported with contralateral hemiparesis including a face of supranuclear type, caused by an infarct of the unilateral ventromedial part of the upper medulla. Data from these patients support the hypothesis that part of the corticobulbar fibres supplying the lower facial muscles descend ipsilaterally in the ventromedial part of the upper medulla and then, after decussation, ascend rostrally to the contralateral facial nucleus. PMID- 9328263 TI - Apolipoprotein E genotype in familial Parkinson's disease. The French Parkinson's Disease Genetics Study Group. AB - APOE genotypes were compared in 57 cases of familial Parkinson's disease, 46 cases of sporadic Parkinson's disease, and 387 controls. The frequency of the APOE allele epsilon4 was similar in patients with Parkinson's disease and controls, but the APOE allele epsilon2, thought to be protective for dementia, was significantly more frequent in patients with sporadic Parkinson's disease than in controls. This is the first study of Parkinson's disease to include familial cases. It confirms the absence of association between the APOE allele epsilon4 and this disease. PMID- 9328264 TI - Transitional progressive multiple sclerosis: a clinical and imaging study. AB - OBJECTIVE: To study the prevalence and the natural course of transitional progressive multiple sclerosis (TPMS). This clinical form is defined by a progressive course beginning many years after an isolated bout. METHODS: 214 consecutive outpatients with definite or probable multiple sclerosis were studied. The prevalence of TPMS was established. Patients with TPMS were compared with patients with other progressive forms of multiple sclerosis according to the clinical course. A prospective one year follow up study was performed in a subgroup of patients to compare progression of the disease using clinical indices and MRI. RESULTS: In this clinical population of 214 outpatients with multiple sclerosis, 55 had secondary progressive multiple sclerosis (SPMS), 38 primary progressive multiple sclerosis (PPMS), and 12 TPMS. Retrospective analysis of the clinical data of these patients shows that TPMS is very similar to SPMS at the beginning of the disease (age at onset, time before progression, clinical symptoms at onset, progression index). In addition a cohort of patients was prospectively followed up clinically and by MRI for one year. CONCLUSIONS: The results did not show any significant differences between the three forms during this follow up. However, all data showed a concordant trend suggesting that at this progressive stage, TPMS is closer to PPMS in terms of progression of disability and new MRI lesions. PMID- 9328265 TI - Toe agnosia in Gerstmann syndrome. AB - The following case report presents a patient exhibiting Gerstmann syndrome accompanied by toe agnosia. A 72 year old right handed woman had a focal lesion in the angular gyrus of the left hemisphere which was caused by a glioblastoma multiforme. The first symptom she had complained of was severe headache. Standardised neuropsychological tests of intelligence, memory, attention, fluency, apraxia, and language functions as well as tests for the assessment of agraphia, acalculia, right-left disorientation, and digit agnosia were performed. The patient displayed all four symptoms of the Gerstmann syndrome--namely, agraphia, acalculia, right-left disorientation, and finger agnosia. The patient did not display aphasia, constructional apraxia, or any other neuropsychological impairment. In addition to the four symptoms of the Gerstmann syndrome an agnosia of the toes was found. Further studies should determine whether finger agnosia in Gerstmann syndrome is usually accompanied by toe agnosia. Finger agnosia in the context of this syndrome may be better named digit agnosia. PMID- 9328266 TI - Isolated myoclonic alien hand as the sole presentation of pathologically established Creutzfeldt-Jakob disease: a report of two patients. AB - Creutzfeldt-Jakob disease may have many atypical presentations before the development of classic progressive dementia and startle myoclonus. In two patients with pathologically established disease association with a progressive alien hand syndrome was the sole initial manifestation of the disease. PMID- 9328267 TI - Magnetic resonance imaging in Creutzfeldt-Jakob disease: evidence of focal involvement of the cortex. PMID- 9328268 TI - Primary central nervous system lymphoma presenting with multiple myeloma-like clinical picture. PMID- 9328269 TI - Attacks of pain in the leg from classic syringomyelia. PMID- 9328270 TI - Anti-GQ1b and anti-GT1a IgG antibodies in a patient with acute demyelinating polyradiculoneuropathy without ophthalmoplegia. PMID- 9328271 TI - Can trauma alone to the trigeminal root relieve trigeminal neuralgia? The case against the microvascular compression hypothesis. PMID- 9328272 TI - About the original description of cerebellar tonsil herniation by Pierre Marie. PMID- 9328273 TI - Blepharospasm induced by flunarizine. PMID- 9328274 TI - Expression of tenascin in astrocytic tumours: too much ado about nothing? PMID- 9328275 TI - Multiple sclerosis associated with duplicated CMT1A: a report of two cases. PMID- 9328276 TI - Distal myasthenia gravis and sensory neuronopathy with anti-50 kDa antibody mimicking sensory-motor neuropathy. PMID- 9328277 TI - Isolation and characterization of Xenopus laevis xSox-B1 cDNA. AB - A family of genes related to the sex-determining region Y gene (SRY) has been termed SOX, and the members are known to encode transcriptional factors and be involved in animal development. We have isolated and sequenced a novel Sox cDNA of Xenopus laevis in this study. The cDNA encodes a protein of 309 amino acids, which encompasses a B-type SOX HMG box; the cDNA was named xSox-B1 in this study. The xSox-B1 mRNA is 1.6 kb in length. Various tissues of adult frog were tested for xSox-B1 mRNA by reverse transcription/ polymerase chain reaction, and the results showed that xSox-B1 expression was highly restricted to the ovary. The message of the xSox-B1 gene was also detected in unfertilized eggs and the late blastula embryos of X. laevis. Recombinant polypeptide of the xSox-B1 HMG domain preferentially binds to the AACAAT sequence. PMID- 9328278 TI - Acarbose and 1-deoxynojirimycin inhibit maltose and maltooligosaccharide hydrolysis of human small intestinal glucoamylase-maltase in two different substrate-induced modes. AB - The inhibition of the glucoamylase-maltase-catalyzed maltose and maltooligosaccharide hydrolysis by acarbose and 1-deoxynojirimycin has been demonstrated. Acarbose and 1-deoxynojirimycin act as potent competitive inhibitors with Ki = 0.8 microM for the hydrolysis of maltose and with Ki values of 0.4 and 0.3 microM, respectively, for the hydrolysis of maltooligosaccharides. In a previous work (Gunther et al., Arch. Biochem. Biophys. 327, 295-302, 1996) using maltitol and maltobionate as inhibitors we were able to discriminate two different binding modes for glucoamylase-maltase: a maltose and an oligosaccharide binding mode. Here we found that structurally quite different substances, namely, the pseudotetrasaccharide acarbose and the monomeric glucose analog 1-deoxynojirimycin, act as competitive inhibitors for maltose and maltooligosaccharide hydrolysis. The Ki values for all used maltooligosaccharides are nearly equal, but for maltose hydrolysis the Ki values are significantly higher by a magnitude factor of two. The differences concerning Ki values can be explained by means of the two-binding-mode model. PMID- 9328279 TI - Purification, characterization, and amino acid sequencing of DNase gamma from rat spleen. AB - An endonuclease named DNase gamma was purified to apparent homogeneity from rat splenocyte nuclei and its properties were characterized. We also determined the NH2-terminal and partial amino acid sequences of the proteolytic internal peptides. The molecular mass of gamma DNase was 33,000 daltons as determined by SDS-polyacrylamide gel electrophoresis. A native molecular mass of 30,000 was estimated by gel filtration. Purified DNase gamma is active in the presence of both Ca2+ and Mg2+ or Mn2+ alone and inhibited by Co2+, Ni2+, Cu2+, and especially Zn2+. Maximal activity was achieved at pH 7.2 in Mops-NaOH buffer. The sequence data on the NH2-terminal and seven internal peptides obtained by sequential digestions with Achromobacter protease I and endoproteinase Asp-N revealed that DNase gamma is a novel endonuclease that shows sequence homology with DNase I. PMID- 9328280 TI - RPE65, the major retinal pigment epithelium microsomal membrane protein, associates with phospholipid liposomes. AB - The retinal pigment epithelium (RPE)-specific protein RPE65 is the major protein of the RPE microsomal membrane fraction. Though RPE65 lacks transmembrane domains or signal peptide, detergents are required for its maximally effective solubilization in isotonic buffers. However, in 0.75-1.0 M KCl, RPE65 is as soluble without detergent, indicating a peripheral membrane association. We wished to understand why this non-membrane-inserted protein was so closely associated with RPE microsomal membranes. To explore the possible involvement of interactions with phospholipids, an isotonic salt-soluble extract of RPE was incubated with phosphatidylcholine (PC)/phosphatidylserine (PS)/phosphatidylinositol liposomes and centrifuged to sediment the liposomes. RPE65 cosedimented with the liposome pellet. RPE65 also cosedimented with synthetic dipalmitoyl-, 1-palmitoyl, 2-docosahexaenoyl-PC or dipalmitoyl-PS liposomes. Incubation with 1 mM Ca2+ or 1 mM EGTA had no effect, indicating a Ca2+-independent association. A spectrophotometric assay showed that this interaction of RPE65 with phospholipid vesicles resulted in increased light scattering, consistent with phospholipid vesicle aggregation. Resonance energy transfer experiments showed that any putative aggregation occurred without subsequent vesicle fusion. This PC affinity was further confirmed by incubation of RPE extract with dimyristoyl-PC-immobilized artificial membrane (IAM.PC) matrix. The RPE65 selectively bound and was elutable with 2% detergent. This RPE65-phospholipid liposome association may explain the solubilization characteristics of RPE65 and may be related to the function of RPE65 and to its physical association with the RPE smooth endoplasmic reticulum. PMID- 9328281 TI - Wheat germ initiation factor 2 (WG x eIF2) decreases the inhibition in protein synthesis and eIF2B activity of reticulocyte lysates mediated by eIF2alpha phosphorylation. AB - Phosphorylation of serine 51 residue in the alpha-subunit of eukaryotic initiation factor 2 (eIF2alpha) impairs the guanine nucleotide exchange (GNE) activity of eIF2B protein and thereby inhibits protein synthesis in mammalian systems, insects and yeast. It is not known if phosphorylation of plant eIF2 can inhibit an eIF2B-like activity. Interestingly purified wheat germ eIF2 (WG x eIF2) can exchange guanine nucleotides in vitro without the addition of any protein factor like eIF2B. It is not clear if this is due to a contaminant eIF2B like activity associated with WG x eIF2 or because the affinity of WG x eIF2 for GDP and GTP is not markedly different. Our observations here indicate that the GNE activity of WG x eIF2 is not inhibited upon phosphorylation of the p41-42 doublet subunit in WG x eIF2 by reticulocyte eIF2alpha kinases, or in the presence of reticulocyte eIF2(alphaP) in which serine 51 residue is phosphorylated. Further, addition of WG x eIF2 reduces the inhibition in eIF2B activity, protein synthesis, and also the formation of 15S complex that occurs between reticulocyte eIF2(alphaP) and eIF2B protein in heme-deficient or poly(IC) treated reticulocyte lysates, presumably by a mechanism of competition between wheat germ and reticulocyte eIF2 for phosphorylation. Unlike reticulocyte eIF2(alphaP), phosphorylated WG x eIF2 is unable to interact with reticulocyte eIF2B to form a 15S complex. The ability of WG x eIF2 to exchange guanine nucleotides independent of an eIF2B like protein and the inability of phosphorylated WG x eIF2 to interact with reticulocyte eIF2B suggests that WG x eIF2 is different from mammalian eIF2 and these differences may have occurred in evolution probably due to some changes in the amino acid sequences around the phosphorylation site in eIF2alpha. PMID- 9328282 TI - Potassium collapses the deltaP in yeast mitochondria while the rate of ATP synthesis is inhibited only partially: modulation by phosphate. AB - Addition of increasing concentrations of K+ to yeast mitochondria in the presence of 0 to 400 microM phosphate and 200 microM Mg2+ led to uncoupled respiration and decreased protonmotive force (deltaP):at 0 K+ deltaP = 213 mV, negative inside, where deltapsi = 180 mV and deltapH = 33 mV, while at 20 mM K+ deltaP = 28 mV, where deltapsi = 16 mV and deltapH = 12 mV. In contrast, the synthesis of ATP resulted in smaller values for the Km and the Vmax in 400 microM Pi and increasing ADP: in 0 K+, Km = 18.6 microM and Vmax = 75.4 nmol (min x mg protein) 1, while in 20 mM K+, Km = 5.2 microM and Vmax = 46.0 nmol (min x mg protein)-1, i.e., when K+ depleted most of the deltaP, and at ADP concentrations below the Km, the rate of ATP synthesis was essentially the same as in the absence of K+. At saturating ADP, the rate of ATP synthesis in the presence of K+ was about 60% of the rate observed without K+. The synthesis of ATP by yeast mitochondria was inhibited by oligomycin or uncouplers. K+ had no effects on rat liver mitochondria. Adenylate kinase activity was much smaller in yeast mitochondria than in rat liver mitochondria and thus did not account for the synthesis of ATP observed in the presence of K+. The effects of K+ on the deltaP of yeast mitochondria were prevented by increasing concentrations of phosphate (1 to 4 mM). At 4 mM phosphate, the deltaP was always above 200 mV and the kinetics of ATP synthesis were as follows: 0 K+ Km = 10.0 microM and Vmax = 88.3 nmol (min x mg protein)-1. At 20 mM K+, Km = 7.4 microM and Vmax = 133 nmol (min x mg protein)-1. PMID- 9328283 TI - Protein modification by lipid peroxidation products: formation of malondialdehyde derived N(epsilon)-(2-propenol)lysine in proteins. AB - Malondialdehyde (MDA), a naturally occurring dialdehyde produced in the membrane lipid peroxidation, is known to react with lysine residues of proteins, but the MDA-lysine adducts generated in the proteins have not been characterized adequately. In the present study, we provide evidence that the enaminal-type MDA lysine adduct, N(epsilon)-(2-propenal)lysine, is formed in human low-density lipoprotein (LDL) upon reaction with MDA or Cu2+. We found that the incubation of N(alpha)-acetyllysine with MDA generated N(alpha)-acetyl-N(epsilon)-(2 propenal)lysine as the predominant product. In addition, a polyclonal antiserum raised against the MDA-modified protein was found to contain antibody populations that could be purified by affinity gel prepared by covalent attachment of N(alpha)-acetyl-N(epsilon)-(2-propenal)lysine. It was concluded that the affinity purified anti-N(epsilon)-(2-propenal) lysine antibody was highly specific to the enaminal derivative of both lysine residues and phosphatidylethanolamine, based on the observations that (i) MDA was the only aldehyde which generated immunoreactive materials in proteins; (ii) among structurally defined MDA-lysine adducts tested, the antibody recognized the enaminal adduct only; and (iii) immunoreactivity to N-(2-propenal)serine was still significant but much weaker than its reactivity to N-(2-propenal)ethanolamine. Furthermore, analysis of antibody recognition sites with a variety of N-(2-propenal)alkylamines revealed that the mono-specific antibody recognized the N-2-propenal-N-ethyl moiety [ (CH2)2-NH-CH=CH-CHO] of enaminal adducts. Determination by a competitive enzyme linked immunosorbent assay demonstrated that N(epsilon)-(2-propenal)lysine accounted for 33.7 and 3.1% of the lysine residues that disappeared during in vitro incubation of LDL with MDA and Cu2+, respectively. These results suggest that N(epsilon)-(2-propenal)lysine represents a major form of MDA covalently attached to proteins. PMID- 9328284 TI - Molecular analysis of carotenoid cyclase inhibition. AB - Later steps of carotenoid biosynthesis catalyzed by cyclase enzymes involve the formation of alpha, beta, and kappa-rings. Examination of the primary structure of lycopene beta-cyclase revealed 55% identity with that of antheraxanthin kappa cyclase. Recombinant lycopene beta-cyclase afforded only beta-carotene, while recombinant antheraxanthin kappa-cyclase catalyzed the formation of beta-carotene from lycopene as well as the conversion of antheraxanthin into the kappa carotenoid capsanthin. Since the formation of beta- and kappa-rings involves a transient carotenoid carbocation, this suggests that both cyclases initiate and/or neutralize the incipient carbocation by similar mechanisms. Several amine derivatives protonated at physiological pH were used to examine the molecular basis of this phenomenon. The beta-and kappa-cyclases displayed similar inhibition patterns. Affinity or photoaffinity labeling using p-dimethylamino benzenediazonium fluoroborate, N,N-dimethyl-2-phenylaziridinium, and nicotine irreversibly inactivated both cyclase enzymes. Photoaffinity labeling using [3H]nicotine followed by radiosequence analysis and site-directed mutagenesis revealed the existence of two cyclase domains characterized by the presence of reactive aromatic and carboxylic amino acid residues. We propose that these residues represent the "negative point charges" involved in the coordination of the incipient carotenoid carbocations. PMID- 9328285 TI - Aryl hydrocarbon (Ah) nonresponsiveness in estrogen receptor-negative MDA-MB-231 cells is associated with expression of a variant arnt protein. AB - Several studies have reported a correlation between expression of the estrogen receptor (ER) and aryl hydrocarbon (Ah) responsiveness in human breast cancer cell lines. MDA-MB-231 cells are ER-negative and Ah-nonresponsive; however, initial studies showed that 2,3,7,8-tetrachlorodibenzo-p-dioxin induced CYP1A1 mRNA levels (5.8-fold) and chloramphenicol acetyltransferase activity (2.6-fold) in high passage (Hp, >50 passages) cells transiently transfected with an Ah responsive plasmid. In contrast, no induction responses were observed in low passage (Lp, <20 passages) cells. The Ah responsiveness of Hp compared to Lp MDA MB-231 cells was associated with a >2-fold increased expression of the Ah receptor in Hp cells. Further analysis revealed that the apparent molecular weight of the Ah receptor mRNA transcript and immunoreactive protein were comparable in Lp MDA-MB-231 and Ah-responsive human HepG2 cells. In contrast, RT PCR analysis of the Ah receptor nuclear translocator (Arnt) protein showed that HepG2 cells expressed the expected 2.6-kb transcript, whereas a 1.3-kb transcript was the major product in MDA-MB-231 cells. Western blot analysis confirmed that HepG2 cells primarily expressed a 97-kDa wild-type form of Arnt, whereas a dominant 36-kDa variant was expressed in MDA-MB-231 cells. Complete sequence analysis of the variant form of Arnt revealed a major deletion of the C-terminal region of the protein (aa 330 to 789). Like HepG2 cells, the wild-type 2.6-kb transcript was detected in ER-positive (Ah-responsive) MCF-7 cells, whereas the low-molecular-weight variant Arnt was dominant in ER-negative MDA-MB-231, MDA-MB 435, and Adriamycin-resistant MCF-7 cells. These results suggest that expression of this protein may be useful as a prognostic factor in breast cancer. PMID- 9328286 TI - Caloric restriction attenuates dityrosine cross-linking of cardiac and skeletal muscle proteins in aging mice. AB - Oxidative damage, particularly to proteins, has been widely postulated to be a major causative factor in the loss of functional capacity during senescence. The nature of the various mechanisms that may contribute to protein oxidation is only partially understood. In this study, concentrations of two markers for oxidative damage, o,o'-dityrosine and o-tyrosine, were determined using stable isotope dilution gas chromatography-mass spectrometry in four tissues of the mouse, namely heart, skeletal muscle, brain, and liver, during youth (4 months old), adulthood (14 months old), and old (30 months old) age. A comparison was made between mice that had access to unlimited calories with those that were restricted to 60% of the caloric intake of the ad libitum regimen. Caloric restriction of this magnitude extends the average and maximum life span of mice by approximately 40%. In vitro studies demonstrated that o,o'-dityrosine was generated selectively in proteins exposed to tyrosyl radical. o-Tyrosine increased in proteins oxidized with hydroxyl radical, which also resulted in a variable increase in o,o'-dityrosine. In mice fed ad libitum, levels of o,o' dityrosine increased with age in cardiac and skeletal muscle but not in liver or brain. In contrast, o-tyrosine levels did not rise with age in any of the tissues examined. These results suggest that tyrosyl radical-induced protein oxidation increases selectively with age in skeletal muscle and heart. Caloric restriction prevented the increase in o,o'-dityrosine levels in cardiac and skeletal muscle but did not influence o-tyrosine levels in any of the four tissues. This selective increase in o,o'-dityrosine levels and its prevention by a life prolonging caloric restriction regimen raise the possibility that oxidation of muscle proteins by tyrosyl radical contributes to the deterioration of cardiac and skeletal muscle function with advancing age. PMID- 9328287 TI - Expression of cytochrome P450 3A7 in Escherichia coli: effects of 5' modification and catalytic characterization of recombinant enzyme expressed in bicistronic format with NADPH-cytochrome P450 reductase. AB - Cytochrome P450 3A7 is the major P450 form present in fetal liver tissue and may be responsible for the detoxification of many drugs that reach the fetal circulation. We report the development of bacterial expression systems for P450 3A7. Maximal yields (up to 50 nmol P450/liter culture) were obtained with a construct in which the 5'-terminus of the 3A7 cDNA was modified to include the MALLLAVFL N-terminal sequence of recombinant bovine P450 17A (H. J. Barnes, M. P. Arlotto, and M. R. Waterman, Proc. Natl. Acad. Sci. USA 88, 5597-5601, 1991) and to incorporate several downstream amino acid substitutions derived from the P450 3A5 sequence. This sequence also appeared optimal for expression of P450 3A4 and 3A5. Recombinant P450 3A7 was partially purified using ion-exchange and hydroxylapatite chromatography and reconstituted with NADPH-cytochrome P450 reductase, cytochrome b5, and lipids. Activity comparable to that of P450 3A4 was demonstrated toward a number of procarcinogens. An alternative approach was used to further characterize recombinant 3A7 due to low yields of recombinant protein in the expression and poor recovery in the purification. P450 3A7 was subcloned into a bicistronic vector containing human NADPH-cytochrome P450 reductase and expressed in bacteria. Recombinant P450 3A7 coexpressed in bacterial membranes with NADPH-cytochrome P450 reductase showed similar levels of activity toward erythromycin (N-demethylation) and ethylmorphine (N-demethylation) to P450 3A4 and 3A5 expressed in the same system, whereas 3A7 was less active toward midazolam (1'- and 4-hydroxylation) and nifedipine (oxidation). PMID- 9328288 TI - Adriamycin and daunomycin semiquinone membrane/buffer partition constants using the spin-broadening technique. AB - Membrane/buffer partition constants have been determined for the semiquinones of adriamycin, Adrsq, and daunomycin, Daunosq, as a function of ionic strength and cholesterol content in phosphatidylcholine multilamellar vesicles using the spin broadening technique. The partition constants corresponding to Adrsq were essentially similar to those of Daunosq under any of the conditions studied. These constants increase with ionic strength increase and decrease with an increase in cholesterol concentration at the membrane. These observations could have implications in the Adr and Dauno cytotoxicities where their corresponding semiquinones are postulated as important intermediates and consequently their interactions with biological membranes should also be considered as important. PMID- 9328289 TI - Further characterization of the major and minor rabbit FMO1 promoters and identification of both positive and negative distal regulatory elements. AB - Previously, two promoters were identified for the rabbit FMO1 gene: a major, upstream promoter (P0) that initiates transcription from exon 0 and a second, minor promoter (P1) located approximately 200 bp downstream and initiating transcription from exon 1. Transcription initiation from the P0 promoter results in elimination of the exon 1 leader sequence from the mature transcript. In this report, we further define the major promoter and identify several positive and negative upstream regulatory domains employing deletion analysis and transient expression in HepG2 cells. Of interest, P0 and P1 were equally active in these assays. A 49-bp fragment spanning position -41 to +8 was found essential for the activity of P0 and also capable of basal transcriptional activity. Interestingly, this same 49-bp region was found necessary for P1 activity. Upstream of P0, three positive regulatory regions (positions -348 to -176, -757 to -584, and -1196 to 829) and two negative regulatory regions (positions -2120 to -1724 and 829 to 757) were identified using deletion mutants. Both P0 and P1 share the most proximate, positive regulatory domain but were regulated differentially by more distal 5' sequences. In addition to the upstream regulatory sequences, a potent negatively acting element was observed within intron 1. Using DNA fragments representing the most potent positive (position -348 to -176) and negative (position -829 to -757) regulatory sequences as probes, we demonstrate the formation of multiple specific DNA/protein complexes with protein factor(s) present in HepG2 nuclear extract. PMID- 9328290 TI - Purification and characterization of thylakoid membrane-bound inorganic pyrophosphatase from Spinacia oleracia L. AB - An inorganic pyrophosphatase (PPase) was purified from thylakoid membrane of spinach leaves to electrophoretic purity by methods including detergent solubilization, ammonium sulfate fractionation, and successive chromatographic techniques. Current protocol yielded about 10% recovery of total activity with a 30-fold purification. The specific activity of the purified enzyme was approximately 400 micromol PPi consumed/mg protein x h. This enzyme is a monomer with a molecular mass of 55 kDa. Several properties, including subunit composition, substrate specificity, ion requirements, inhibitor sensitivities, and amino acid composition, have been studied. Mg2+ is an essential cofactor for the thylakoid PPase. The preferred substrate for the hydrolytic reaction of PPase appears to be dimagnesium pyrophosphate. K+ could not stimulate the enzymatic activity of thylakoid PPase, while F- was a potent inhibitor. Group-specific modification of the thylakoid PPase demonstrates possible involvement of carboxylate residues in the enzymatic activity. Furthermore, antibodies raised against thylakoid PPase in a rabbit could inactivate the PPi hydrolysis of thylakoid and the purified enzyme, but not that of vacuolar H+-PPase, indicating both PPi hydrolases are structurally distinct. PMID- 9328291 TI - Expression cloning of a novel farnesylated protein, RDJ2, encoding a DnaJ protein homologue. AB - The CAAX farnesyltransferase is a heterodimeric enzyme that attaches a farnesyl group to a single cysteine in cellular proteins which terminate in the sequence CAAX, where C is cysteine, A is an aliphatic amino acid, and X is most often methionine or serine. Substrates include the p21ras proteins, nuclear lamins, and a series of retinal proteins. To date, a limited number of substrates for the farnesyltransferase have been identified, predominantly by demonstration of the attachment of a farnesyl group to previously identified cDNA clones which encode proteins containing an appropriate carboxyl-terminal tetrapeptide. We describe here the use of a cDNA fusion protein expression library, together with enzymatic in vitro [3H]farnesyl radiolabeling, as a means of identifying novel farnesylated proteins. One candidate cDNA was fully cloned and found to be a homologue of the Escherichia coli heat shock gene dnaJ. The predicted amino acid sequence of this protein was found to terminate with the tetrapeptide Cys-Ala-His-Gln, which conforms to the consensus sequence for recognition by farnesyltransferase, and was shown to undergo in vivo farnesylation. This farnesylated protein, designated RDJ2 (rat DnaJ homologue 2), is a novel and ubiquitously expressed DnaJ homologue and is the newest member of the subfamily of DnaJ-related proteins which are posttranslationally modified by protein farnesylation. PMID- 9328292 TI - Expression and purification of recombinant rhinovirus 14 3CD proteinase and its comparison to the 3C proteinase. AB - Human rhinovirus (HRV) is a positive-stranded RNA virus with an open reading frame that encodes for a single polyprotein of about 3000 amino acids. The HRV polyprotein is proteolytically processed; eight of nine cleavages are catalyzed by the 3C and/or the 3CD proteinases. We have expressed and purified recombinant HRV14 3C and 3CD proteinases and investigated their substrate selectivity and inhibitor sensitivity. Expressed 3CD proteinase had the P1/P1' residues of the 3C/3D cleavage site mutated from Gln/Gly to Ala/Ala in order to prevent autocleavage. The 3CD proteinase activities were measured by utilization of native, chromogenic, and fluorogenic peptide substrates. The 3CD proteinase exhibited < or =15% activity, compared to 3C, toward peptidyl p-nitroanilide substrates which contain only the p-nitroaniline moiety in the prime side. The 3C and 3CD proteinases exhibited similar activities for both internally quenched fluorogenic and native peptides. These results suggest that the two enzymes have similar but slightly different substrate specificity, especially on their preference for prime side residues. Inhibitor sensitivities toward classical proteinase inhibitors were generally similar for both enzymes. Small peptidyl inhibitors, specifically designed and synthesized for HRV14 3C, also inhibited the 3CD proteinase. Taken together, our data indicate that the 3D domain of 3CD proteinase had some influence on substrate recognition, but did not have dramatic impact on its interaction with inhibitors. PMID- 9328293 TI - Elevated ferritin production, iron containment, and oxidant resistance in hemin treated leukemia cells. AB - Hemin (ferriprotoporphyrin IX), the oxidized prosthetic group of hemoglobin, is a source of potentially cytotoxic iron, but in chronic low doses can induce cytoprotection against iron-stimulated oxidative stress. The latter property of hemin has been examined, using murine L1210 cells and three different oxidant generating systems: (i) glucose/glucose oxidase, (ii) near-ultraviolet irradiation, and (iii) dye-mediated photodynamic action. Cells treated with the lipophilic iron donor ferric-8-hydroxyquinoline, Fe(HQ)2 (1 microM, 30 min) were found to be more sensitive to oxidative killing than nontreated controls. However, cells challenged after long-term (20-24 h) exposure to hemin (10 microM) were substantially more resistant than controls and were sensitized far less by Fe(HQ)2. Immunoblot analyses of 24-h hemin-treated cells indicated that the ferritin heavy (H) subunit was elevated 12- to 15-fold, whereas the light (L) subunit was essentially unchanged. Experiments carried out with 55Fe(HQ)2 showed that iron uptake capacity of cells was greatly enhanced after hemin treatment. More specifically, hemin-stimulated cells were found to contain approximately 9 times more immunoprecipitable ferritin iron after incubation with saturating levels (4-5 microM) of 55Fe(HQ)2 and approximately 3 times more iron per ferritin molecule compared with nonstimulated controls. The nonferritin iron content of the latter was estimated to be approximately 40 times greater than that of the former following low-level (0.5 microM) 55Fe(HQ)2 treatment. These results are consistent with the idea that induced ferritin, enriched in H-chain, sequesters redox active iron rapidly and copiously, thereby enhancing cellular resistance to oxidants. PMID- 9328294 TI - Two Sp sites are important cis elements regulating the upstream promoter region of the gene for rat type I hexokinase. AB - Multiple transcriptional start sites have been identified for the gene encoding the rat Type I isozyme of hexokinase (White, J.A., Liu, W., and Wilson, J. E., Arch. Biochem. Biophys. 335, 161-172, 1996); these are clustered at positions approximately -460, -300, and -100 relative to the translational start codon (ATG, with A being +1). PC12 cells and H9c2 cells were transfected with luciferase reporter constructs containing genomic sequence between positions 3366 and -171. Marked (85%) decrease in promoter activity was associated with deletion of sequence between -742 and -516. In DNase I footprinting experiments, two regions, called P1 (-552 to -529) and P2 (-480 to -458) boxes, were protected by proteins present in nuclear extracts from PC12 cells. Mutation or deletion of the P2 box had no effect on promoter activity; protection in this region, which includes the most upstream cluster of transcriptional start sites, is attributed to binding of RNA polymerase II or associated factors. In contrast, mutations or deletions in the P1 box had markedly detrimental effects on promoter activity and on binding of proteins in PC12 cell nuclear extracts. Maintenance of a consensus Sp1 binding site centrally located in the P1 box was critical for both promoter activity and binding. A second Sp1 site (-570), just upstream from the P1 box, was also shown to be functionally important but no protection of this region was detected in footprinting experiments, presumably reflecting lower affinity at this site under the conditions used. Supershift experiments demonstrated the involvement of Sp1, Sp3, and Sp4 in formation of complexes with the P1 box region and implicate these transcription factors in regulating promoter activity associated with this region. Another series of reporter constructs, including sequence between -171 and -1, permitted detection of an additional promoter activity downstream from -364. While not yet extensively characterized, it is already evident that the cis elements influencing the downstream promoter activity are distinct from the Sp factors determined to be important in expression from the upstream promoter region. PMID- 9328295 TI - Structural diversity among subtypes of small-conductance Ca2+-activated potassium channels. AB - 125I-Apamin and photolabile derivatives of the toxin have been used to investigate the binding properties and subunit composition of small conductance Ca2+-activated potassium channels (SK(Ca) channels) expressed on plasma membranes from rat brain, rabbit liver, or rat pheochromocytoma (PC12) cells. On all preparations, 125I-apamin recognized single classes of acceptor binding sites with similar high affinity (Kd approximately 3-6 pM). Gallamine, however, was found to readily discriminate between 125I-apamin acceptors present in these preparations, showing a maximal approx nine-fold difference in affinity for acceptors expressed by rabbit liver or PC12 cells. Affinity-labeling patterns revealed the expression of different hetero-oligomeric combinations of high (86 or 59 kDa) and low (33 or 30 kDa) molecular mass 125I-apamin-binding polypeptides, consistent with pharmacological differences. Alternative expression of either 86- or 59-kDa polypeptides appeared to be the most important factor influencing gallamine's affinity for SK(Ca) channel subtypes. Both high- and low molecular-mass polypeptides are integral membrane proteins, the latter being glycosylated in a tissue-specific manner. PMID- 9328296 TI - Progesterone and testosterone hydroxylation by cytochromes P450 2C19, 2C9, and 3A4 in human liver microsomes. AB - Roles of human cytochrome P450 (P450 or CYP) 2C9, 2C19, and 3A4 in the oxidation of progesterone and testosterone were studied in recombinant P450 enzymes and in human liver microsomes. In vitro inhibition experiments showed that progesterone and its 17alpha- and 21-hydroxylated metabolites and 11-deoxycortisol suppressed the CYP2C19-dependent R-warfarin 7-hydroxylation activities, with progesterone being the most active. These steroid chemicals also inhibited CYP2C9-dependent S warfarin 7-hydroxylation activities though lesser extents seen with those in CYP2C19 enzyme. Progesterone was found to be a competitive inhibitor of CYP2C19 and CYP2C9 in human liver microsomes. Recombinant CYP2C19 catalyzed progesterone to form 21-hydroxyprogesterone as a major product and 16alpha- and 17alpha hydroxyprogesterone as minor products. CYP2C9 also had progesterone 21 hydroxylation activities, although the activities were lower than those catalyzed by CYP2C19. Vmax/Km ratios for the progesterone 21-hydroxylation activity of CYP2C19 were determined to be 13- and 32-fold higher than those of CYP2C9 and 3A4, respectively. CYP3A4 oxidized progesterone to form 16alpha-, 6beta-, and 2beta-hydroxyprogesterone as major products and 21-hydroxyprogesterone as a minor product, but did not produce detectable levels of 17alpha-hydroxyprogesterone. Immunoinhibition experiments suggested that anti-CYP2C9 (which inhibits both CYP2C9 and CYP2C19 catalytic activities) suppressed the progesterone 21 hydroxylation activities catalyzed by liver microsomes of humans and monkeys and that anti-CYP2C11 inhibited the progesterone 21-hydroxylation activities catalyzed by liver microsomes of male rats. CYP2C19 was also found to oxidize testosterone at 17-position to form androstenedione. Androstenedione formation catalyzed by liver microsomes of humans and monkeys and of male rats was suppressed by anti-CYP2C9 and anti-CYP2C11, respectively. These results suggest that CYP2C19 plays important roles in the oxidation of progesterone and testosterone in human liver microsomes, although the physiological significance of these metabolic pathways remains unclear. CYP2C9 may have some, but lesser extent than those by CYP2C19, of the catalytic roles for the metabolism of progesterone and testosterone by human liver microsomes. PMID- 9328297 TI - Factors associated with gallstone disease in the MICOL experience. Multicenter Italian Study on Epidemiology of Cholelithiasis. AB - The epidemiological associations of gallstone disease were evaluated in a general population sample of 29,584 individuals (15,910 men and 13,674 women; age range, 30-39 years) belonging to 14 cohorts examined between December 1984 and April 1987. Subjects were screened for the presence of gallstones by gallbladder ultrasonography, completed a questionnaire, and underwent a physical examination and blood chemistry tests. Participants were considered to have gallstone disease if they had already had cholecystectomy or gallstones. Statistical associations were established by univariate analysis of the age-standardized data and by stepwise multiple logistic regression. Increasing age and body mass index and a maternal family history of gallstone disease were the most consistent associations (both at univariate and multivariate analysis and in both sexes) found in this study. Personal history of dieting was associated with gallstone disease in men, and at univariate analysis, in women. Decreasing serum total cholesterol levels and increasing serum triglycerides were associated with gallstone disease in both sexes in the multivariate analysis. In women, associations were also found with a number of pregnancies and paternal family history of gallstone disease. A slight but negative association with contraceptive pill use was identified only at multivariate analysis. Associations (investigated at univariate analysis) were also found with diabetes, cirrhosis, angina or myocardial infarction, and peptic ulcer. There was no association with smoking habits and use of aspirin or antirheumatic drugs. PMID- 9328298 TI - Multivesicular stellate cells in primary biliary cirrhosis. AB - Stellate cells have only recently received attention in patients with primary biliary cirrhosis (PBC). We have used electron microscopy and morphometry to perform a qualitative and quantitative examination of lipid-storing activity of stellate cells in liver biopsies of 26 patients with noncirrhotic and cirrhotic PBC. In parallel with this study, a comparative analysis of the morphology of stellate cells in 51 patients with livers of normal histology was performed. There was a marked increased in the total number of lipid vesicles in stellate cells in all PBC patients when compared with livers with normal histology. Multiple multivesicular stellate cells were seen in the livers of 21 of 26 patients with PBC. There were 11 to 28 lipid vesicles per multivesicular stellate cell in sizes of 1 microm to 5 microm in diameter per lipid vesicle. Hepatocytes showed little or no steatosis in 24 of 26 (92%) PBC patients. Multivesicular stellate cells were not seen in female patients with normal liver histology. These results suggest that there is an alteration in hepatic lipid storage that involves stellate cells in PBC that could be an early manifestation of this disease. Its significance remains to be elucidated. PMID- 9328299 TI - Purification of a newly identified alkaline sphingomyelinase in human bile and effects of bile salts and phosphatidylcholine on enzyme activity. AB - The hydrolysis of sphingomyelin (SM) generates important signals regulating cell proliferation and apoptosis. Acid and neutral sphingomyelinases (SMase) have been identified and their biological effects intensively studied. We recently found in human bile a novel alkaline SMase that may have important roles in hepatobiliary diseases. In this work, we purified the enzyme and studied the factors influencing enzyme activity. Purification steps included Sephadex G25, diethylaminoethyl (DEAE)-Sepharose, Sephacryl S-200, and sphingosylphosphorylcholine (SPC) affinity chromatographies. A single protein of 92 kd was obtained with the specific enzyme activity increased 1,154-fold. The enzyme specifically hydrolyzed SM to ceramide, had a weaker activity against phosphatidylcholine (PC), and no activity against either phosphatidylethanolamine (PE) or p-nitrophenyl phosphate. Its optimum pH was 9.0 and its Vmax and Km were 45 micromol/h/mg and 2.5 x 10(-5) mol/L, respectively. The enzyme activity was dependent on concentration and structure of bile salts. Both trihydroxy and dihydroxy bile salts at concentrations up to their critical micellar concentrations activated the alkaline SMase, trihydroxy bile salts being more potent than dihydroxy ones. The side chain of trihydroxy bile salts was also important. Taurocholate (TC) was most effective in activating SMase, followed by glycocholate (GC), and cholate. 3-((3-cholamidopropyl)dimethylammonio) propanesulfonate (CHAPS) alone had no effect on SMase activity but inhibited TC induced activation of SMase. PC competitively inhibited bile alkaline SMase activity, with the 50% inhibition occurring at a PC/SM ratio of approximately 28. In conclusion, we purified a novel alkaline SMase from human bile and found that its activity is dependent on the levels of two major biliary components: PC and bile salts. PMID- 9328300 TI - Sustained gallbladder stasis promotes cholesterol gallstone formation in the ground squirrel. AB - Although gallbladder stasis exists in most patients with cholesterol gallstones, it is unknown whether stasis is a causative factor of gallstone disease or merely a consequence of it. We studied the impact of sustained gallbladder stasis induced by a cholecystokinin (CCK)-A receptor antagonist (MK-329) on gallstone formation in ground squirrels fed either a trace or a high-cholesterol diet. MK 329 markedly inhibited gallbladder contraction in vitro in response to CCK (at EC100, control: 3.6 +/- 0.5 vs. MK-329: 1.1 +/- 0.3 g; P < .05) and increased gallbladder fasting volume in vivo (control: 462 +/- 66 vs. MK-329: 1,004 +/- 121 microL; P < .05). Whereas the high-cholesterol diet alone (1%-cholesterol diet + placebo) increased the cholesterol saturation index (CSI) in control animals (trace-cholesterol diet + placebo), MK-329 significantly (P < .05) decreased the CSI in both hepatic and gallbladder bile in animals on the trace-(trace cholesterol diet + MK-329) as well as on the high-cholesterol diets (1% cholesterol diet + MK-329). The mucin content of the mucus layer on the epithelial surface of the gallbladder wall more than doubled (P < .05) with the high-cholesterol diet; adding MK-329 to the latter group produced a further 82% increase (P < .05). The cholesterol diet + MK-329 group had the highest (100%) incidence of cholesterol crystals that were evident in fresh gallbladder bile, coincident with a shortened nucleation time (2.5 +/- 0.6 days; P < .05 vs. the cholesterol diet + placebo group, 5.8 +/- 1.0 days or the other 2 groups, >21 days). Bile from animals on the trace-cholesterol diet, whether or not receiving MK-329, lacked crystals in bile and exhibited a normal nucleation time (>21 days). Thus, stasis per se may lower the CSI, but its detrimental effect on the gallbladder predominates locally, and so accelerates cholesterol crystal formation in this model. PMID- 9328301 TI - Diencephalic and cerebellar pathology in alcoholic and nonalcoholic patients with end-stage liver disease. AB - Formalin-fixed sections from the brains of 36 patients (30 alcoholic and 6 nonalcoholic) with autopsy-proven cirrhosis who died while in a hepatic coma were stained with hematoxylin and eosin, and examined for the presence of diencephalic, cerebellar, pontine, and basal ganglia lesions. Significant neuropathology was identified in 23 of 36 cases consisting of mammillary body and thalamic lesions characteristic of Wernicke encephalopathy (WE) (9 cases, all alcoholic patients) and cerebellar degeneration (20 cases, 17 alcoholic and 3 nonalcoholic patients). Clinical diagnosis of WE had been entertained during life in only 2 of these patients. All cases, alcoholic and nonalcoholic, manifested mild to severe Alzheimer's type II astrocytosis. No cases of central pontine myelinolysis nor acquired (non-Wilsonian) hepatocerebral degeneration were found. These findings show that the brains of a high proportion of cirrhotic patients with end-stage liver disease manifest concomitant unsuspected diencephalic and cerebellar pathology. The high incidence of WE underscores the need for early sustained treatment of alcoholic cirrhotic patients with vitamin B1. Evaluation of the neurological sequelae of liver transplantation, particularly of alcoholic patients with end-stage liver disease, may require a careful neurological and radiological assessment both before and after surgery. PMID- 9328302 TI - Exhaled nitric oxide and oxygenation abnormalities in hepatic cirrhosis. AB - Impaired arterial oxygenation, ranging from increased alveolar-arterial oxygen gradient (AaDo2) to hypoxemia, is commonly present in patients with cirrhosis. Nitric oxide (NO), through pulmonary vasodilatation, may play a major role in the oxygen abnormalities of cirrhosis. Our aim was to study the relationship between NO production and O2 abnormalities in 45 nonsmoking patients with cirrhosis and without major cardiovascular and respiratory diseases. Intrapulmonary shunting was detected by contrast-enhanced (CE) echocardiography. Lung volumes and diffusion, arterial blood gas analysis, serum NO2-/NO3-, NO output in the exhaled air, and cardiac index by the echocardiographic method were determined in all patients. Twenty-seven (60%) patients had an abnormally increased (> 15 mm Hg) AaDo2. The mean values of exhaled NO output and serum NO2-/NO3- were significantly higher in cirrhotic patients than in controls (252 +/- 117 vs. 75.2 +/- 19 nL/min/m2, P < .0001; and 47.5 +/- 29.4 vs. 32.9 +/- 10.1 micromol/L, P < .02, respectively). In all patients, there was a significant correlation between exhaled NO and AaDo2 (r = .78, P < .0001). Twelve patients (26.6%) were found to have CE-echocardiographic evidence of intrapulmonary shunting (positive CE-echo). Nine patients were considered to have hepatopulmonary syndrome (HPS) on the basis of an AaDo2 > 15 mm Hg and positive CE-echo. These 9 patients had a mean value of exhaled NO significantly higher than patients without HPS (331 +/- 73.2 vs. 223 +/- 118.4 nL/min/m2, P < .05). In all patients, cardiac index was positively correlated with exhaled NO (r = .47, P < .001) and with serum NO2-/NO3- (r = .43, P < .01). The results suggest an important role of NO in the oxygenation and circulatory abnormalities of patients with cirrhosis. PMID- 9328303 TI - Donor type microchimerism is an infrequent event following liver transplantation and is not associated with graft acceptance. AB - Donor-type microchimerism, the presence of a minority population of donor-derived haematopoietic cells following solid organ transplantation, has been postulated as a mechanism for induction of donor-specific graft tolerance. The stability, frequency, and relevance of microchimerism with respect to long-term outcome, however, remains uncertain. Using a polymerase chain reaction (PCR)-based method of microsatellite analysis of highly polymorphic short tandem repeat sequences (STRs) to detect donor-type cells, DNA from 11 patients was analyzed prospectively at specific time points for 12 months following liver transplantation, and from a further six patients retrospectively 2 years after liver transplantation. Using a panel of STRs, transient peripheral blood donor microchimerism was detected in 2 of 11 patients at a single time-point following transplantation, but persistent evidence of donor-derived cells was not observed during the study period. Analysis of DNA extracted from skin and duodenum in two patients likewise failed to show donor-type cells at these sites. None of the six patients in the retrospective arm showed donor microchimerism, resulting in an overall detection rate of 1.58%. These results suggest that donor microchimerism following liver transplantation is an infrequent event, and that the generation of graft tolerance is independent of microchimerism. PMID- 9328304 TI - A randomized clinical trial of ursodeoxycholic acid as adjuvant treatment to prevent liver transplant rejection. AB - Acute rejection following orthotopic liver transplantation is a common problem despite current immunosuppressive regimens. Ursodeoxycholic acid (UDCA) has been shown in small, open-labeled studies to prevent rejection episodes, although its effects on complications such as infections, length of hospital stay, and survival have not been evaluated. We conducted a randomized, placebo-controlled, double-blind trial to determine if UDCA (10-15 mg/kg/d) added to a cyclosporine based immunosuppressive regimen was associated with a decrease in the incidence of at least one episode of acute cellular rejection. Secondary end-points included determining differences in the total number of rejection episodes, the use of muromonab-CD3, the incidence of infections, length of hospital stay, and survival at 90 days and 1 year. Fifty-two patients were randomized, 28 to the treatment group and 24 to the placebo group. During the 3 months of the trial, there was no difference between the placebo and UDCA groups in the number of patients who were rejection-free; however, there were significantly fewer patients in the treatment group who had multiple episodes of acute rejection (0 vs. 6; P = .007). Patients in the treatment group experienced a significantly lower incidence of bacterial infections (4% vs. 29%; P = .02), shorter hospital stay (25 days vs. 34 days; P = .03), and better 90-day survival (100% vs. 83%; P = .04) and 1-year survival (93% vs. 79%). The addition of UDCA to a cyclosporine based immunosuppressive regimen results in significantly fewer patients experiencing multiple episodes of rejection and improved survival at 90 days and at 1 year. The use of UDCA as adjuvant therapy for patients undergoing liver transplantation who are treated with a cyclosporine-based immunosuppressive regimen should be considered. PMID- 9328305 TI - Renal functional reserve and nitric oxide in patients with compensated liver cirrhosis. AB - To investigate the role of nitric oxide (NO) with respect to kidney function and liver cirrhosis, we evaluated renal function, as well as cyclic guanosine monophosphate (cGMP) and NOx (nitrite/nitrate [NO3-/NO2-]) as indirect markers of NO formation in plasma and urine at rest and during amino acid (aa)-induced glomerular hyperfiltration in patients with Child A liver cirrhosis and portal hypertension (n = 12), and in healthy controls (n = 10). Baseline filtration rate (GFR) and effective renal plasma flow (ERPF) were significantly lower in patients with cirrhosis than in controls (GFR: mean 88 +/- SD 16 mL/min vs. 106 +/- 15 mL/min, P = .01, ERPF: 477 +/- 93 vs. 561 +/- 72 mL/min, P = .002). In both groups amino acid (aa) infusion increased GFR, ERPF, as well as cGMP and urinary NOx. Changes in GFR were similar in cirrhotic patients and controls (28.3% +/- 14% in cirrhotics and 26% +/- 11% in controls), but the degree of aa-induced changes in ERPF was more marked in patients with liver cirrhosis (31.8% +/- 17% vs. 18.6% +/- 12%, P = .02). Plasma levels of NOx and cGMP were similar in either group at baseline and during aa infusion, whereas NOx and cGMP excretion in cirrhotics was constantly 14% to 24% lower than in the control group. We conclude that patients with compensated liver cirrhosis and portal hypertension already have an impaired kidney function. In addition our data suggest a cirrhosis related dissociation between ERPF and GFR during aa stimulation. Further studies are warranted to find out whether a local imbalance between vasoconstrictors and vasodilators, e.g., decreased local NO formation, plays a key role for this phenomenon. PMID- 9328306 TI - Aplastic anemia complicating orthotopic liver transplantation. AB - The clinical characteristics and outcome of posttransplantation aplastic anemia (AA) were determined in 12 of 1,736 patients (0.007%) undergoing orthotopic liver transplantation (OLT) that were afflicted with AA. None of the affected patients had a history of hematologic disease. Median patient age was 53 years (range, 2 61 years); 10 of the affected patients were men, and 2 were women. The etiologies of AA included non-A, non-B, non-C fulminant hepatic failure (FHF) (3 patients), graft-versus-host disease (4 patients), Parvovirus-induced (1 patient), and idiopathic (4 patients). The median duration between OLT and the onset of AA was 12 days (range, 11-14 days) in the 3 patients undergoing OLT for FHF; in contrast, AA developed in the other 9 patients at 37 days (range, 27-51 days) after OLT. Eleven patients were treated with reduction of their cyclosporine or tacrolimus dosage, granulocyte colony-stimulating factor, anti-thymocyte globulin, and Solumedrol. Two of the 3 patients developing AA following OLT for FHF achieved hematologic recovery 21 and 92 days after diagnosis. In contrast, all 9 non-FHF patients developing AA after OLT died, 5 due to infectious complications and 4 following intracranial bleeding. AA is an unusual complication of OLT. In the setting of FHF, it affects young males in the early posttransplantation period, and, when infectious complications can be avoided, remission and stable allograft function can be anticipated. However, in the non FHF patient, AA occurs in older individuals later in the posttransplantation period and has a uniformly poor outcome. PMID- 9328307 TI - Oral glutamine challenge in cirrhotics pre- and post-liver transplantation: a psychometric and analyzed EEG study. AB - Latent or sub-clinical hepatic encephalopathy is a recognized complication of cirrhosis and is thought to represent one end of the spectrum of neuropsychiatric impairment, which occurrs as a result of portal-systemic shunting. We studied the psychometric, analyzed electroencephalography (EEG), and venous blood ammonia responses to an oral glutamine challenge in 17 patients with cirrhosis and in 4 normal controls. The cirrhotics were attending for liver transplant assessment and had no clinical evidence of hepatic encephalopathy. The oral glutamine challenge was repeated following liver transplantation. Five of sixteen patients (31%) showed impaired performance on at least one of the baseline psychometric tests. There was a correlation between fasting venous ammonia and choice reaction time (r = .7, P < .01). Following glutamine challenge there was a significant increase in blood ammonia from a mean fasting value ranging between 58 micromol/L to 120 micromol/L (P < .01), between significant prolongation of reaction times of 387 ms to 428 ms (P < .01), and an increase in mean EEG amplitude between 68.5 microV to 78.6 microV (P < .001). Four normal controls who were challenged with glutamine and 6 cirrhotic patients who were challenged with water showed no change in any of these parameters. Following orthotopic liver transplantation (OLT) the eight patients studied had normal baseline psychomotor performance with significant improvements in digit symbol, digit span, information processing, number connection tests (P < .05), and reaction time (P < .005). Posttransplantation, there were no significant changes in blood ammonia, analyzed EEG, or choice reaction time in response to oral glutamine challenge (six patients). We conclude that short lived changes in blood ammonia (in cirrhotics) can cause significant impairment of sensitive tests of brain function and that psychometric performance is improved following OLT. PMID- 9328309 TI - Bcl-2 is overexpressed and alters the threshold for apoptosis in a cholangiocarcinoma cell line. AB - Cholangiocarcinoma is a malignant neoplasm originating from cholangiocytes. The mechanisms responsible for oncogenesis of cholangiocytes are unknown. Resistance to apoptosis, especially by altered expression of B-cell lymphoma/leukemia 2 (Bcl 2) family members, has been implicated as a mechanism contributing to malignant transformation. Thus, our aim was to test the hypothesis that altered expression of Bcl-2 family members by cholangiocarcinoma cells renders them resistant to apoptosis. We compared the apoptotic threshold and expression of the Bcl-2 protein family members, Bcl-2, Bcl-XL, and Bax, in two human cell lines: 1) nonmalignant human cholangiocytes immortalized by transfection with the simian virus 40 (SV 40) large T antigen; and 2) a malignant human cholangiocarcinoma cell line. Apoptosis was induced pharmacologically using beauvericin. Bcl-2, Bcl x long, and Bax protein expression were evaluated by immunoblot analysis, and Bcl 2 expression was modulated using antisense technology. The cholangiocyte and malignant/nonmaligant phenotype of both cell lines was verified using both in vitro and in vivo approaches. Beauvericin induced apoptosis of nonmalignant cholangiocytes in a concentration- (0 to 25 micromol/L) and time- (0 to 6 hours) dependent manner. In contrast, malignant cholangiocytes were resistant to apoptosis. Although expression of Bcl-x long and Bax protein were similiar in the two cell lines, Bcl-2 protein expression was 15-fold greater in malignant than in nonmalignant cholangiocytes. An 18 mer bcl-2 antisense oligonucleotide reduced expression of Bcl-2 protein by 50% and increased the rate of beauvericin-induced apoptosis more than threefold in the malignant cells. Our results support the hypothesis that resistance to apoptosis by overexpression of Bcl-2 may be a feature of cholangiocarcinoma. PMID- 9328308 TI - Treatment of fibrolamellar hepatoma with subtotal hepatectomy or transplantation. AB - Fibrolamellar hepatoma (FL-HCC) is an uncommon variant of hepatocellular carcinoma (HCC), distinguished by histopathological features suggesting greater differentiation than conventional HCC. However, the optimal treatment and the prognosis of FL-HCC have been controversial. Follow-up studies are available from 1 year to 27 years, after 41 patients with FL-HCC were treated with partial hepatectomy (PHx) (28 patients) or liver transplantation (13 patients). In this retrospective study, the effect on outcome was determined for the pTNM stage and other prognostic factors routinely recorded at the time of surgery. Cumulative survival at 1, 3, 5, and 10 years was 97.6%, 72.3%, 66.2%, and 47.4%. Tumor-free survival at these times was 80.3%, 49.4%, 33%, and 29.3%. The TNM stage was significantly associated with tumor-free survival. Patients with positive nodes had a shorter tumor-free survival than those with negative nodes (P < .015). Patient survival was most adversely affected by the presence of vascular invasion (P < .05). FL-HCC is an indolently growing tumor of the liver, which usually was diagnosed in our patients at a stage too advanced for effective surgical treatment of most conventional HCC. Nevertheless, long-term survival frequently was achieved with aggressive surgical treatment. When a subtotal hepatectomy could not be performed, total hepatectomy (THx) with liver transplantation was a valuable option. PMID- 9328310 TI - Evidence for the polyclonal nature of focal nodular hyperplasia of the liver by the study of X-chromosome inactivation. AB - Focal nodular hyperplasia (FNH) of the liver is a benign tumor commonly considered as a reactive disorder related to a pre-existing vascular malformation. However, the pathogenesis of this lesion has been recently discussed. To determine whether FNH is a polyclonal or a clonal lesion, we investigated the inactivation pattern of the X chromosome, using molecular genetic analysis of the DNA methylation pattern at a polymorphic site on the human androgen receptor gene (HUMARA). Fifteen FNH were studied, and results were compared with those obtained from 7 hepatic adenomas (HA) and 2 hepatocellular carcinomas (HCC). DNA was extracted from both lesional and nonlesional livers, fixed, and paraffin-embedded. To assess the methylation pattern, we used a quantitative fluorescent polymerase chain reaction (PCR) procedure that allows for the accurate measurement of the peak intensities of each allele. Three patients were noninformative because they were homozygous at the HUMARA locus. According to the threshold for monoclonality established by a titration curve, all FNH showed a random pattern of X-chromosome inactivation consistent with a polyclonal lesion. In contrast, all but 1 hepatic adenoma and all hepatocellular carcinomas were clonal, as shown by the nonrandom pattern of X-chromosome inactivation observed in these cases. In conclusion, these results suggest that FNH should be considered as a reactive disorder rather than as a tumoral proliferation. Discordant results recently observed in the literature could be at least in part explained by methodological differences in the PCR procedure. PMID- 9328311 TI - Extracellular matrix remodeling at the early stages of liver regeneration in the rat. AB - Previous studies have shown that activity of urokinase-type plasminogen activator (u-PA) increases very rapidly (within 1 minute) after partial hepatectomy. In view of the well-recognized roles of u-PA as one of the major initiators of the matrix proteolysis cascade and as an activator of plasminogen and hepatocyte growth factor (HGF), we studied matrix degradation in liver shortly after partial hepatectomy. The activation of plasminogen to plasmin following partial hepatectomy was examined by Western blot analysis, and a small increase in plasmin at approximately 15 minutes followed by a large elevation at approximately 3 to 6 hours after partial hepatectomy was detected. In addition, we found that fibrinogen, the major substrate for plasmin, begins to be degraded at approximately 15 to 30 minutes following partial hepatectomy. Using immunohistochemical staining, we detected that the distribution of fibrinogen in normal liver is localized to the perisinusoidal space surrounding the periportal region. A decreased distribution of fibrinogen in the periportal region was found by 15 minutes and continued through 24 hours following partial hepatectomy. In addition, the distribution of fibronectin in normal liver was localized to the perisinusoidal space surrounding the periportal and the pericentral regions. A strikingly decreased distribution of fibronectin in the periportal region was found at 5 minutes after partial hepatectomy. Furthermore, we observed that the protein levels of laminin, entactin, and fibronectin in an extracellular matrix (ECM)-enriched preparation decreased shortly after partial hepatectomy, and were restored later. No changes were observed with either vitronectin or the integrin chain alpha(v). In contrast to the protein levels of the ECM components, the messenger RNA (mRNA) levels of fibronectin, integrin chain beta1, and integrin chain alpha(v) gradually increased over 18 hours and then decreased thereafter. Taken together, these results suggest that rapid reorganization of selected ECM components are important for hepatocyte proliferation at the early stages of liver regeneration. PMID- 9328312 TI - Localization and cellular sources of activins in normal and fibrotic rat liver. AB - Activins are dimeric proteins, members of the transforming growth factor beta (TGF-beta) gene superfamily, consisting of the beta-subunits of inhibin (betaA and betaB). Recently, it was shown that activin A (betaA:betaA) inhibits DNA replication and induces apoptosis in rat parenchymal cells in vitro and in vivo. Cryostat sections of normal livers and livers of rats treated with intraperitoneal injections of CCl4 were stained for the different inhibin subunits and desmin, a marker for stellate cells (SC). Staining for inhibin-alpha was invariably negative, both in normal and fibrotic rat liver. In normal liver, inhibin-betaA subunit immunoreactivity was localized in parenchymal cells (PC). Staining for inhibin-betaB was weaker but similarly distributed. In fibrotic livers, connective tissue septa were strongly immunoreactive for inhibin-betaA. Desmin-positive stellate cells (SC) accumulated in areas strongly immunoreactive for inhibin-betaA and several cells were positive for both desmin and inhibin betaA. Staining for inhibin-betaB was weaker but similarly distributed. As above data pointed to a possible role for PC and SC, we examined by Northern blot analysis, the expression of inhibin-alpha, -betaA, and -betaB messenger RNA (mRNA) in total RNA extracted from freshly isolated SC and PC of normal and CCl4 treated liver and in cultured SC. Inhibin-betaA mRNA was predominantly expressed in PC of normal liver. Expression was lost in PC of CCl4-treated liver. Inhibin betaB mRNA expression was induced in SC of CCl4-treated liver. Inhibin-betaA mRNA, and to a lesser extent, inhibin-betaB mRNA expression was rapidly induced in cultured SC. The presence of activin A in conditioned media of cultured SC was shown by Western blotting. Apoptotic cells, identified by terminal deoxy transferase mediated X-dUTP nick end labeling (TUNEL)-staining, were found predominantly in and near the fibrotic septa. IN CONCLUSION: 1) while activin A was constitutively expressed in PC of normal liver, its expression was lost in PC of fibrotic liver; 2) expression of activins was induced in activated SC; and 3) apoptotic cells were found predominantly near the septa, in support of the hypothesis that activin A, derived from activated SC in the septa, contributes to the induction of cell death in neighboring PC. PMID- 9328313 TI - Retinoids exacerbate rat liver fibrosis by inducing the activation of latent TGF beta in liver stellate cells. AB - Liver stellate cells (SCs) play central roles in both the storage of retinol and the development of liver fibrosis. The present study is aimed to understand the mechanism by which retinoic acid (RA, an active metabolite of retinol) enhances hepatic fibrosis in rats. We tested the effect of 9-cis-RA on several aspects in vitro rat SC cultures, including the activity of cellular plasminogen activator (PA), messenger RNA (mRNA), and protein levels of transforming growth factor-beta (TGF-beta) mRNA level of type-I procollagen, and the activity of type-I collagenase. Employing the rat liver fibrosis model produced by porcine serum, we also estimated the effect of oral administration of a stable RA analog on the progression of the fibrosis, as well as on hepatic TGF-beta contents. In vitro SC cultures, 9-cis-RA enhanced cellular PA and plasmin levels thereby induced plasmin-mediated activation of latent TGF-beta. Active TGF-beta generated self stimulated its synthesis as well as that of collagen and suppressed the production of collagenase in an autocrine manner. In in vivo rat models, an RA analog accelerated the porcine serum-induced fibrosis by enhancing TGF-beta contents and, thus, collagen levels in the liver, although the RA analog alone was not fibrogenic. These results suggest that RA exacerbated liver fibrosis, at least in part, by inducing the activation and production of latent TGF-beta in liver SCs. PMID- 9328314 TI - Evidence of a role for matrix metalloproteinases in cold preservation injury of the liver in humans and in the rat. AB - Previous studies have determined that proteases are important in cold preservation injury to the liver. The purpose of this study was to determine the role of matrix metalloproteinases (MMPs) in cold preservation injury. Effluents were collected from rat livers after various periods of preservation either in Eurocollins solution or in University of Wisconsin (UW) solution. Effluents were also collected from 17 human donor livers stored in UW solution. To determine whether sinusoidal endothelial cells released MMPs when placed in the cold, these cells were isolated from rat livers and cultured at 4 degrees C. Gelatin zymography, quantitative assay of gelatinolytic activity, immunoprecipitation, and Western blotting were used to identify metalloproteinases and to measure their activity. Human and rat liver effluents contained gelatin-digesting bands on zymography. Their appearance was inhibited by specific metalloproteinase inhibitors and also by lactobionate, the major ingredient of UW solution. The most prominent bands in humans and the rat appeared at approximately 72 kd and 92 kd, suggesting that they were the MMPs 72-kd gelatinase and 92-kd gelatinase. Supernatants of isolated rat sinusoidal endothelial cells stored in the cold contained similar bands. In the rat, the proteinases were present in both latent and active forms, but, in humans, predominately the latent form was seen. In humans, there were four prominent bands in the gelatin zymography. By immunoprecipitation, two of the bands were identified as the 92-kd gelatinase and a dimer or polymer of 92-kd gelatinase. Using Western blotting with a monoclonal antibody, a third band was identified as 72-kd gelatinase. In quantitative terms, gelatinolytic activity increased with time of cold storage in humans and in the rat. In the rat, gelatinolytic activity was greater when Eurocollins was the preservative than when UW solution was used. Taken together, these results indicate an important role for MMPs in the injury produced by cold preservation of the liver. PMID- 9328315 TI - Flow cytometric analysis of vesicular pH in rat hepatocytes after ethanol administration. AB - We examined the effect of ethanol administration on intravesicular pH in intact hepatocytes by applying a flow cytometric technique to detect fluorescein isothiocyanate-dextran (FITC-dextran) in acidic vesicles. Rats were pair-fed liquid diets containing either ethanol or isocaloric carbohydrate for 1 to 5 weeks. Our study showed that ethanol administration increased the in situ pH of hepatic lysosomes by 0.15 to 0.2 pH units. This pH increase was sufficient to cause a significant reduction in lysosomal protein degradation. Long-term ethanol administration also caused a significant alkalinization of hepatic endosomes, and this increased pH was sustained over the course of vesicular acidification in hepatocytes incubated in vitro. Direct exposure of hepatocytes from rats fed control diet to either 25 mmol/L ethanol or 50 micromol/L colchicine also brought about a rapid alkalinization of acidic vesicles in a manner that resembled that seen in hepatocytes from ethanol-fed rats. These same treatments augmented the vesicular alkalinization already present in cells from ethanol-fed animals. Although ethanol administration had no effect on the content of the hepatic mannose-6-phosphate/IGFII receptor, the results indicate that sustained alkalinization of endosomes could have important functional consequences by impairing M-6-P/IGFII receptor recycling, thereby disrupting the delivery of newly synthesized hydrolases to lysosomes. This decreased complement of hydrolases within lysosomes together with alkalinization of the intralysosomal compartment would result in an overall decrease in lysosomal proteolysis. PMID- 9328316 TI - Cycloheximide prevents apoptosis, reactive oxygen species production, and glutathione depletion induced by transforming growth factor beta in fetal rat hepatocytes in primary culture. AB - We have previously reported that transforming growth factor-beta (TGF-beta) induces apoptosis in fetal hepatocytes in primary culture. This effect was found to be associated with an increase in intracellular reactive oxygen species (ROS) and a lowering of total cellular reduced glutathione (GSH). In this study, we investigated whether protein synthesis plays a role in these TGF-beta-induced effects. When fetal hepatocytes were incubated in the presence of cycloheximide, a specific protein synthesis inhibitor, TGF-beta-induced apoptosis was completely blocked. The overall intracellular oxidized state of the cells, when measured using either 2',7'-dichlorofluorescein diacetate (DCFH) by laser-scanning confocal microscopy, or both DCFH and hydroethidine (DHE) by flow cytometric analysis, was increased transiently after the addition of TGF-beta. This increase was abolished by incubation of the cells in the presence of cycloheximide. Furthermore, the decrease in the total cellular GSH content induced by TGF-beta in these cells was not observed when cycloheximide was present. Cycloheximide effect was not associated with an enhancement of cysteine and restoration of cellular glutathione level, because inhibition of GSH synthesis with buthionine sulfoximine (BSO) did not prevent the cycloheximide protective effect. Experiments performed to check whether Fas (APO-1) ligand might be the protein that needs to be synthesized have indicated that this possibility can be excluded. All these findings suggest that apoptosis elicited by TGF-beta in fetal hepatocytes requires the synthesis of an unknown protein before ROS production, glutathione loss, and oxidative stress. PMID- 9328317 TI - Critical role of Fas/Fas ligand interaction in CD28-independent pathway of allogeneic murine hepatocyte rejection. AB - Cytolytic induction of T cells requires both the T-cell receptor (TCR)-mediated antigenic stimulation and the CD28-mediated co-stimulatory signal. Blockade of the interactions between CD28 and its ligands, CD80 and CD86, prolongs the survival of allografts in some transplantation models. However, we found that allogeneic hepatocytes were completely rejected within 7 days after intrasplenic transplantation, even when treated with monoclonal antibodies (mAbs) against CD80 and CD86 (anti-CD80/86). Recent studies have shown that there are two main mechanisms of T-cell-mediated cytotoxicity, perforin-based and Fas-based ones. It has been shown that the liver is highly sensitive to induction of apoptosis by an agonistic anti-Fas mAb. We then investigated the role of the Fas/Fas ligand (FasL) system in the CD28-independent allogeneic hepatocyte rejection. With the anti-CD80/86 mAb treatment, hepatocytes from C57BL/6 lpr/lpr (B6 lpr) mice, which express little Fas antigen, could survive for 7 days after intrasplenic transplantation, and hepatocytes from C57BL/6 (B6) mice could also survive for 7 days in the spleen of C3H/ He gld/gld (C3H gld) mice, which express no functional FasL. CD28-independent induction of cytotoxicity against allogeneic hepatocytes was not observed when the effector cells were derived from C3H gld mice. These results indicated that the Fas/FasL system plays a critical role in the CD28 independent pathway of allogeneic hepatocyte rejection. PMID- 9328318 TI - Transient immunosuppression with FK506 permits long-term expression of therapeutic genes introduced into the liver using recombinant adenoviruses in the rat. AB - The host immune response limits the duration of expression of adenovirally transduced genes and precludes long-term gene expression upon re-administration of the virus. In this study we wished to evaluate whether short-term immunosuppression of the host, at the time of recombinant virus administration, would allow expression of the therapeutic gene product upon virus reinjection. Gunn rats were used as recipients of recombinant adenoviruses expressing human BUGT (Ad-hBUGT) or E. coli beta-galactosidase (Ad-LacZ). Rats were treated with FK506 (1-1.5 mg/kg, per OS daily) for three days beginning 24 hours before each virus injection. Control groups did not receive any immunosuppressant. The serum bilirubin level was reduced from 7.1 +/- 0.75 mg/dL to 2.0 +/- 0.7 mg/dL within two days of viral injection in both FK506 treated and control groups, and then gradually increased in 6 weeks. FK506-treated rats had low or undetectable antibody titers against the recombinant adenovirus and minimal or no cytotoxic T lymphocyte (CTL) response against adenovirus-infected cells. The tolerized rats received two subsequent injections 42 and 98 days after the first injection, which reduced the bilirubin levels again to 2.0 +/- 0.56 and 2.2 +/- 0.61 mg/dL, respectively. In contrast, control rats developed high titer neutralizing antibodies and a CTL response, and their serum bilirubin levels were not reduced following subsequent injections. We conclude that short-term FK506 treatment around the time of virus administration prevents the host immune response, permitting long-term gene therapy by repeated administration of the recombinant virus. PMID- 9328319 TI - Chlormethiazole inhibition of cytochrome P450 2E1 as assessed by chlorzoxazone hydroxylation in humans. AB - Chlormethiazole is a sedative and anticonvulsive drug used in the treatment of alcohol withdrawal. Because it had been reported that chlormethiazole inhibits the alcohol-inducible cytochrome P450 2E1 in rat liver, we investigated the in vivo and in vitro effect of this drug on cytochrome P450 2E1 in human beings. The activity of this cytochrome was assessed using chlorzoxazone as a probe. The 6 hydroxychlorzoxazone-chlorzoxazone blood concentration ratio, reflecting the cytochrome P450 2E1 activity, was determined in 10 controls and in 24 alcoholic patients who had entered a hospital for detoxification. Alcoholic patients were administered either chlormethiazole (1.3-2.3 g/d) or chlorazepate (100-300 mg/d) as a sedative. Cytochrome P450 2E1 activity was significantly increased in alcoholic patients treated with chlorazepate (1.16 +/- 0.40 vs. 0.27 +/- 0.03, P < .05). In contrast, chlormethiazole treatment inhibited chlorzoxazone hydroxylation almost totally (0.046 +/- 0.03, P < .001). After 7-14 days of ethanol withdrawal, alcoholic patients treated with chlorazepate had ratio values similar to those of controls (0.31 +/- 0.05), whereas values from alcoholic patients treated with chlormethiazole remained low (0.049 +/- 0.01) even though chlormethiazole doses were gradually decreased. Pharmacokinetic studies in controls showed that chlormethiazole-mediated inhibition was present even when chlormethiazole was not detectable in the blood. In addition, the effect of chlormethiazole on cytochrome P450 2E1 was studied in vitro using human liver microsomes. Dixon plot analyses showed a noncompetitive inhibition (Ki = 12 micromol/L). These data clearly show that chlormethiazole is an efficient inhibitor of chlorzoxazone metabolism and thus of cytochrome P450 2E1 activity in human beings. Because cytochrome P450 2E1 induction after chronic ethanol consumption has detrimental effects on the liver through free radical formation, treatment of alcohol detoxification with chlormethiazole may be beneficial. PMID- 9328320 TI - Differences in hepatic processing of dietary and intravenously administered copper in rats. AB - The biliary pathway represents the major excretory route for copper (Cu). It has been suggested that glutathione (GSH) plays a role in this process. However, biliary secretion of endogenous Cu is unaffected in canalicular multispecific organic anion transporter (cmoat)/multi-drug resistance protein (mrp2)-deficient GY/TR- rats, which is a mutant rat strain expressing defective canalicular adenosine triphosphate (ATP)-dependent GSH-conjugate transport and which is unable to secrete GSH into bile. Secretion of Cu after iv Cu load is markedly impaired in GY/TR- rats when compared with normal Wistar (NW) rats. Administration, iv, of 65, 325, or 2300 nmol/100 g body wt CuSO4 dose-dependently increased Cu secretion in normal Wistar (NW) rats. Secretion rates in GY/TR rats were much lower and plateaued with higher loads at a level of about 35 nmol/h/100 g body wt. Clearance of an intravenous (iv) bolus of 64Cu (250 nmol/100 g body wt) was faster in GY/TR- rats than in controls, but secretion of 64Cu into bile was clearly reduced in the mutants. Specific activity of biliary Cu was similar in both groups. To investigate the removal of excess dietary Cu via bile, GY/TR and NW rats received water supplemented with Cu (CuSO4 8 mmol/L) for up to 12 weeks (Cu-fed) or tap water (controls). Cu feeding resulted in an increase of biliary Cu secretion from approximately 6 to approximately 30 nmol/h/100 g body wt within two weeks, both in NW and GY/TR- rats; Cu secretion also did not further increase during the course of the experiment. Hepatic Cu content was similar in NW and GY/TR- rats and progressively increased during Cu feeding. Our data indicate that biliary secretion of diet-derived Cu proceeds exclusively via a saturable Cu transporting system, which is distinct from cmoat/mrp2 and which is independent of biliary GSH. This transport may be mediated by the recently identified Cu-ATPase. In contrast, excess hepatic Cu after iv Cu load depends on cmoat/mrp2 activity for rapid removal. It is concluded that iv administered and dietary (endogenous) Cu is, in part, processed differently by rat liver, which might be related to differences in Cu redox state. PMID- 9328321 TI - Additive inhibitory effect of hydrocortisone and cyclosporine on low-density lipoprotein receptor activity in cultured HepG2 cells. AB - Both glucocorticoids and cyclosporine are used to prevent rejection in organ transplant recipients. However, long-term treatment with these drugs is known to induce hyperlipidemia and premature development of atherosclerosis. In previous studies, we have shown that the immunosuppressive drug cyclosporine inhibits catabolism of low-density lipoproteins (LDL) mainly by reducing the expression of LDL-receptor messenger RNA (mRNA), thus explaining the increased plasma levels of LDL cholesterol observed in patients treated with cyclosporine. In the present study, our objective was to investigate the mechanism by which glucocorticoids increase plasma levels of LDL cholesterol. We studied the catabolism of LDL in the human hepatoma cell line HepG2. Our results show that hydrocortisone at physiologically relevant concentrations inhibits LDL binding, uptake, and degradation in a dose-dependent way. Moreover, hydrocortisone also reduces the expression of LDL-receptor mRNA in a dose-dependent way. Cyclosporine also has an additive inhibitory effect on hydrocortisone in the catabolism of LDL. The 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor fluvastatin reverses the inhibitory effect of both hydrocortisone and cyclosporine. We conclude that treatment with hydrocortisone and/or cyclosporine induces increased plasma levels of LDL cholesterol because of reduced hepatic LDL receptor activity. HMG-CoA reductase inhibitors reverse this undesirable effect and thus reduce the risk of the development of atherosclerosis in patients subjected to immunosuppressive treatment. PMID- 9328322 TI - Effect of moderate exercise on insulin sensitivity and substrate metabolism during post-exercise recovery in cirrhosis. AB - We examined whether a single bout of moderate exercise has a beneficial effect on insulin sensitivity and fuel homeostasis in cirrhosis. Clinically stable cirrhotic patients and age-, sex-, and weight-matched controls participated in insulin clamp studies (either euglycemic hyperinsulinemic or hyperglycemic hyperinsulinemic) in combination with indirect calorimetry and [6,6-2H2]glucose. Three to seven days later, studies were repeated following a single bout of exercise (30 minutes of treadmill exercise at 60% of maximal aerobic capacity). After an overnight fast, following exercise, both cirrhotic and control individuals showed a shift in fuel utilization to enhanced lipid oxidation, decreased glucose oxidation, and increased nonoxidative glucose disposal rates (i.e., glycogen synthesis in muscle) when compared with pre-exercise rates but differences were statistically significant only in the patient group. During euglycemic hyperinsulinemia, insulin-mediated glucose disposal was significantly reduced in cirrhotic patients (3.43 +/- 0.26 vs. 7.36 +/- 0.48 mg/kg/min, P < .01). Following exercise, glucose uptake increased significantly in cirrhotic patients when compared with pre-exercise levels (P < .05) but remained unchanged in the control group. The increase in total body glucose disposal in cirrhotic patients was entirely accounted for by an increase in nonoxidative glucose disposal (0.81 +/- 0.20 vs. 0.51 +/- 0.15 mg/kg/min, P < .05). During combined hyperglycemia/hyperinsulinemia, however, insulin sensitivity was unaffected by exercise in both patients and control individuals. In summary, in cirrhotic patients, a single bout of moderate exercise 1) causes a shift in substrate utilization with an increase in lipid oxidation in the postexercise period that is significantly more pronounced than in controls, and 2) increases insulin sensitivity only during euglycemia but not during the more physiological condition of hyperglycemia. Single bouts of moderate exercise therefore may not have a beneficial effect on the metabolic status of patients with chronic liver disease. PMID- 9328323 TI - Induction of cMrp/cMoat gene expression by cisplatin, 2-acetylaminofluorene, or cycloheximide in rat hepatocytes. AB - The human multidrug-resistance-associated protein (MRP), a member of the adenosine triphosphate (ATP)-binding cassette transporter superfamily, is frequently overexpressed in tumor cells resistant to antineoplastic drugs. In the rat, two Mrp isoforms have been identified, Mrp and cMrp. cMrp, also called Mrp2 or cMoat (canalicular multispecific organic anion transporter), is expressed in the canalicular membrane of rat hepatocytes and mediates the excretion of glucuronate, sulfate, and glutathione conjugates into bile. We investigated the expression of cMrp and Mrp in rat hepatocytes in primary culture. Treatment with the chemical carcinogen 2-acetylaminofluorene (2-AAF), the antineoplastic drug cisplatin, and the protein-synthesis inhibitor cycloheximide led to a dose dependent and time-dependent increase in cmrp gene expression. A 347-base pair cmrp complementary DNA (cDNA) probe served to demonstrate the induction of cmrp messenger RNA (mRNA) with 40 micromol/L 2-AAF, 5 micromol/L cisplatin, or 5 micromol/L cycloheximide. An analogous response was obtained for the increase in cMrp protein. Mrp mRNA was below the detection limit in Northern blots of RNA from liver and hepatocyte cultures, in contrast to rat testis mRNA which served as a positive control. Immunofluorescence microscopy of cultured hepatocytes was used to visualize cMrp in the plasma membrane. Treatment with 2-AAF led to a marked increase in the immunofluorescence signal confirming the cMrp-inducing potency of 2-AAF. In conclusion, the inducing effect of the compounds studied may reflect a general inducibility of hepatic cMrp by a variety of cytotoxic, carcinogenic, and chemotherapeutic agents which is likely to be of relevance for the acquisition of multidrug resistance during chemotherapy and in the process of chemical carcinogenesis in the liver. PMID- 9328324 TI - Clinical and family studies in genetic hemochromatosis: microsatellite and HFE studies in five atypical families. AB - A candidate gene (HFE) has been described for hereditary hemochromatosis on chromosome 6. The study of well-defined atypical hemochromatosis families using genetic markers may increase our understanding of the sensitivity and the specificity of genotyping in hemochromatosis. One hundred and thirteen Canadian families with genetic hemochromatosis were surveyed to find atypical families as possible examples of people with genetic recombinations. All families underwent clinical investigations including iron studies and HLA typing. Each individual was typed at three polymorphic microsatellite loci (D6S105, D6S1260, and D6S299) on chromosome 6. Sixteen subjects were studied for the two missense mutations described for the candidate gene for hemochromatosis (C282Y, H63D). There were eight HLA-identical siblings found in four different families (five men, three women; age range 30-72) with normal transferrin saturation and ferritin levels. There were two patients identified who were homozygous for the C282Y mutation without biochemical evidence of iron overload, and two patients with no evidence of the mutation with significant iron overload. Our conclusions are as follows: 1) finding HLA-identical siblings without iron overload does not confirm a genetic recombination, 2) difficulties in phenotypic definition of disease and the description of new iron overload syndromes that may differ from classical genetic HC cause complicated genetic studies, and 3) finding iron-loaded patients without a C282Y mutation and patients that are homozygous for the C282Y mutation without evidence of iron overload may limit the use of genotyping in population screening for hemochromatosis. PMID- 9328325 TI - Identification and functional characterization of the promoter region of the human organic anion transporting polypeptide gene. AB - The organic anion transporting polypeptide (OATP) of the basolateral hepatocyte membrane mediates multispecific uptake of anionic and other amphipathic substrates from sinusoidal blood plasma. To investigate the mechanisms controlling OATP expression, the 5'-flanking region of the human OATP gene was isolated from a P1-derived artificial chromosome genomic clone. Sequence analysis of the OATP promoter showed a number of consensus binding sites for both ubiquitous and liver-enriched transcription factors. Transfection of HepG2 cells with a series of 5'-deleted promoter-luciferase constructs identified the minimal promoter region within 91 base pairs relative to the transcription initiation site. A putative silencer element was localized in the -662/-440 region. The minimal promoter was also active in Chang liver, Madin-Darby canine kidney, and Chinese hamster ovary cells, indicating that basal promoter function is independent of liver-specific regulatory mechanisms. In transfected HepG2 cells, taurocholate (100 micromol/L) stimulated and triiodothyronine (1 micromol/L) inhibited OATP promoter activity, whereas hydrocortisone, dexamethasone, beta estradiol, estrone-3-sulfate, and testosterone had no significant effect. Reverse transcription polymerase chain reaction analysis showed an increase in OATP messenger RNA in the livers of four patients with chronic cholestatic liver disease compared with three noncholestatic controls. The up-regulation of OATP expression by taurocholate could serve to enhance the sinusoidal efflux of toxic intracellular compounds during chronic cholestasis. PMID- 9328326 TI - Spontaneous and iatrogenic spreading of liver-derived cells into peripheral blood of patients with primary liver cancer. AB - The prognosis for patients with primary liver cancer (PLC) often depends on tumor recurrence and the development of extrahepatic metastases, particularly after liver transplantation. We have developed a sensitive test to detect both spontaneous circulation of tumor cells and the spread of liver cells due to chemoembolization and alcoholization. Reverse-transcription polymerase chain reaction was used to search for cells expressing alpha-fetoprotein (AFP) messenger RNA in the peripheral blood of 84 patients with PLC and 102 controls (55 patients with chronic hepatitis and/or cirrhosis, 10 patients with benign liver tumors or liver metastases from intestinal cancers, and 37 healthy individuals). By spiking the blood of healthy volunteers with HepG2 cells, we assessed the sensitivity limit: one HepG2 cell mixed with 10(7) leukocytes. All 102 controls tested negative. In contrast, 28 patients (33.3%) with PLC tested positive. Positivity for the test was significantly associated with portal thrombosis, tumor size, intravascular tumor emboli, serum AFP level, and extrahepatic metastases. Patients were followed up for a mean period of 39 +/- 51 weeks: the probability of developing extrahepatic metastases was significantly higher in positive than in negative patients. Eighteen negative patients with PLC were tested before, 1 hour after, and 24 hours after locoregional therapy: 9 tested positive either 1 or 24 hours after alcoholization or chemoembolization. In conclusion, we have developed a highly specific and sensitive test to detect circulating tumor cells in patients with PLC. This test is likely to be clinically useful in evaluating the risk of developing extrahepatic metastases and the possibility of iatrogenic spreading of liver-derived, possibly tumorous, cells. PMID- 9328327 TI - Prevalence, risk factors, and genotype distribution of hepatitis C virus infection in the general population: a community-based survey in southern Italy. AB - In 1996 the prevalence, risk factors, and genotype distribution of hepatitis C virus (HCV) infection were assessed in the general population of a town in southern Italy. The sample was selected from the census by a systematic 1:4 sampling procedure. The participation rate was 96.6%. Among the 1,352 subjects enrolled, 195 (14.4%) tested reactive to antibody to HCV (anti-HCV) with enzyme immunoassay (EIA 3). When further tested with recombinant immunoblot assay (RIBA 3), 170 subjects (87.2%) tested positive, 23 subjects (11.8%) had indeterminate results, and 2 subjects (1%) tested negative. Thus, the overall anti-HCV EIA positive RIBA-confirmed prevalence was 12.6% (170 of 1,352 subjects) and increased from 1.3% in subjects younger than 30 years to 33.1% in those > or =60 years of age. This latter age group accounted for 72.3% of all anti-HCV-positive subjects. Females tested positive more frequently than males (14.1% vs. 10.5%; P < .05). Alanine transaminase (ALT) concentrations were abnormal in only 4.1% (7/170) of anti-HCV EIA-positive RIBA-confirmed subjects. This suggests that ALT screening is not useful in the detection of anti-HCV-positive subjects in a general population. The results of multiple logistic regression analysis showed that an age of less than 45 years, the use of glass syringes, and dental therapy were all independent predictors of anti-HCV positivity. HCV RNA was detected by polymerase chain reaction in 75.9% of the 195 anti-HCV EIA-positive subjects: in 84.7% (144/170) of the RIBA-confirmed subjects; in 17.4% (4/23) tested as RIBA indeterminate; and in neither of the two subjects who tested RIBA negative. HCV type 1b was detected in 75 subjects (50.7%), type 2b in 1 subject (0.7%), type 2c in 66 subjects (44.6%), type 3a in 4 subjects (2.7%), and type 4 in two subjects (1.3%). These figures differ from those of Italian patients with chronic liver disease in whom genotype 2 is more rare. None of the individuals was infected with more than one genotype. The distribution of the two most common HCV viral types (1b and 2c) was not statistically different in terms of mean age, sex, or risk factors and suggests that they may have had a parallel spread in this community. These findings provide one of the highest overall anti-HCV prevalence rates in a general population with a likely cohort effect, i.e., decreased risk of infection along generations. These observations may indicate an epidemic or focus of hepatitis C that occurred several years earlier. The majority of anti HCV-positive subjects in the oldest age group and with no clinical evidence suggests that HCV infection is a very prolonged and indolent disease. PMID- 9328328 TI - Virological response to interferon treatment in hepatitis C virus carriers with normal aminotransferase levels and chronic hepatitis. AB - Hepatitis C virus (HCV) carriers with normal aminotransferase levels often show histological chronic hepatitis. This study was undertaken to determine the effect of interferon (IFN) in such patients. Nineteen HCV carriers with normal aminotransferase activities and chronic hepatitis were randomized to receive IFN alpha2b (3 million units 3 times weekly for 12 months) or no treatment. Therapy was monitored by qualitative and quantitative determination of viral RNA. Patients who did not clear HCV RNA after 6 months discontinued therapy. In all, 9 patients constituted the control group, while 10 patients were treated. Five of these patients, still viremic after 6 months, stopped IFN. The remaining 5 patients, who cleared the viral RNA within 6 months, completed the 12-month course. Three of these patients relapsed off treatment, and 2 were still free of viremia 12 months after stopping therapy. A transient flare-up of aminotransferase activities was detected in 2 patients during treatment and in 3 patients after. None of the 9 control patients cleared the viral RNA during follow-up. A variable degree of sequence heterogeneity was detected in the hypervariable region before therapy, and IFN treatment decreased sequence diversity in all patients. These results indicate that IFN therapy can be effective in chronic HCV carriers with normal aminotransferase activities, inducing short-term virological response in 3 of 10 patients and sustained response in 2. The effects of treatment on viral load and quasispecies complexity were similar to those reported previously in patients with increased aminotransferase activities. PMID- 9328329 TI - Liver failure in children with hepatitis A. AB - There have been very few reports dealing with liver failure related to hepatitis A in children. Moreover, the criteria usually used for selecting patients with fulminant hepatitis A for liver transplantation have not been evaluated in children. Therefore, the current study was conducted retrospectively in a single French urban pediatric liver transplantation center to serve as a reminder of the potential severity of hepatitis A in children and to identify predictors of outcome. Children were selected by chart review using a data base system and were grouped according to outcome for analyses purposes. Over a 15-year period, 24 children with hepatitis A showed evidence of liver failure, including 6 children who did not develop hepatic encephalopathy, 7 children in whom encephalopathy occurred but resolved spontaneously, and 11 children in whom death or liver transplantation was the outcome. The mean age at onset was 6.5 years. Those with the most rapid onset of liver failure from onset of jaundice had the best chance of recovery without developing encephalopathy. Otherwise, no predictive factors of outcome were found at onset of liver failure. Among the 18 children who developed encephalopathy, the best early prognostic indicator of a poor outcome irrespective of the grade of encephalopathy, appeared to be a prothrombin time level below 21% of normal combined with a serum bilirubin level above 400 micromol/L. Therefore, these two prognostic indicators may be helpful in deciding liver transplantation in children with hepatitis A-induced fulminant liver failure. PMID- 9328330 TI - HLA-C genes and susceptibility to type 1 autoimmune hepatitis. AB - Susceptibility to autoimmune hepatitis (AIH) is associated with the HLA A1-B8-DR3 haplotype, DR4 antigen, and, more specifically, the HLA DRB3*0101, DRB1*0301, and DRB1*0401 alleles. Few investigators, however, have examined the HLA C locus in AIH, which warrants detailed study in view of its recently described roles in immunoregulation. Eighty-seven adult, white patients with well-characterized type 1 AIH and 100 controls were studied. HLA C and HLA DRB1 alleles were assigned by polymerase chain reaction (PCR)-based genotyping. HLA A and B antigens were determined by standard microlymphocytotoxicity assay. Extended haplotypes were constructed according to known patterns of linkage disequilibrium. Only one HLA C locus allele, Cw*0701, which was present in 54% of patients versus 34% of controls (P = .006; relative risk [RR] = 1.54) was associated with AIH. The overall increase in the frequency of the Cw*07 gene (70.1% of patients vs. 54% of controls; P = .024; RR = 1.3) was due entirely to inheritance of the Cw*0701 allele rather than the other Cw*07 alleles, Cw*0702, *0703, and *0704. The RR for Cw*0701 (RR = 1.54) is greater than that for HLA A1 (RR = 1.33) and DRB1*0301 (RR = 1.49), but less than that for HLA-B8 (RR = 1.75). The present findings suggest that the gene or genes conferring susceptibility to AIH lie in the region centromeric to the HLA A locus between HLA C and DRB1. Although linkage disequilibrium with both B8 and DRB1*0301 may account for our finding of an increased frequency of Cw*0701, it is also possible that this allele contributes to disease susceptibility, perhaps by interaction with natural killer cells or cytotoxic T lymphocytes. PMID- 9328331 TI - Molecular features of the hepatitis B virus nucleocapsid T-cell epitope 18-27: interaction with HLA and T-cell receptor. AB - The strength of the cytotoxic T lymphocyte (CTL) response is believed to influence the final outcome of hepatitis B virus (HBV) infection. Among the different CTL epitopes so far identified, the sequence 18-27 of the HBV nucleocapsid antigen is widely recognized by CTL of HLA-A2-positive patients with acute self-limited HBV infection, and represents the main component of a peptide based therapeutic vaccine aimed at stimulating the antiviral CTL response in patients with chronic hepatitis B. In the present study, we further analyzed the features of this important HBV region by the following: 1) defining the contribution of individual residues of the epitope to the interaction with the T cell receptor (TCR) and with the HLA-A0201 molecule; 2) assessing the antigenicity of this viral region in the context of the different HLA-A2 subtypes; and 3) testing whether this sequence can stimulate not only HLA-class I but also HLA class II restricted T-cell responses. A clear hierarchy was observed in the ability of individual residues to act as TCR or HLA binding sites. Furthermore, the sequence HBc18-27 was able to be recognized by specific CTL when presented in the context of different HLA-A2 subtypes. Finally, this HBV region was also found to stimulate HLA class II restricted T-cell responses. These data further increase the potential coverage and efficacy of therapeutic vaccines based on the HBc18-27 sequence. PMID- 9328332 TI - Correlation of biochemical response to interferon alfa with histological improvement in hepatitis C: a meta-analysis of diagnostic test characteristics. AB - The current goal of interferon treatment for chronic hepatitis C is to normalize alanine aminotransferase (ALT) and to eradicate detectable viral RNA. Many patients do not achieve this objective during treatment, and most do not sustain these outcomes after interferon is discontinued. However, biochemical or virological responses to interferon may not reflect accurately the histological consequences of therapy. The aim of this study was to determine the extent to which the biochemical measures reflect the histological outcomes in the treatment of hepatitis C with interferon alfa using a meta-analysis of diagnostic test characteristics. The data sources were English and non-English language studies retrieved from Medline (from 1966 to December 1995). The study selection included studies in which interferon alfa was used for treatment of chronic hepatitis C with liver biopsies performed before and after therapy. Data on histological and biochemical outcomes were extracted independently by two reviewers. Two separate criteria were used for defining histological response. When strict definitions of histological improvement were considered, histology improved in 28% (95% confidence interval [95% CI], 17%-43%) of patients after interferon treatment. The sensitivity and specificity of the ALT for determining histological change were 70% (95% CI, 56%-81%) and 66% (95% CI, 56%-75%), respectively. As many as 17% (95% CI, 9%-30%) of subjects with an abnormal ALT at the end of therapy may have improved histologically after interferon therapy. When less stringent definitions of histological improvement were considered, 62% (95% CI, 51%-72%) improved after therapy. The sensitivity and specificity of the ALT for determining histological change were 55% (95% CI, 44%-65%) and 75% (95% CI, 67% 81%), respectively. As many as 51% (95% CI, 38%-64%) may have improved, despite failure to normalize ALT. A substantial number of patients may improve histologically after interferon therapy. The significance of histological changes observed after interferon therapy must be weighed against the limitations of liver biopsy and the uncertain natural history of hepatitis C. Nevertheless, the ALT does not always reflect liver histology accurately after interferon alfa treatment and may underestimate histological improvement. PMID- 9328333 TI - Isolation and molecular characterization of hepatitis B virus X-protein from a baculovirus expression system. AB - The X protein (HBx) of the human Hepatitis B Virus (HBV) is a regulatory protein that exercises a transcriptional activator function on a variety of regulatory elements and is therefore considered to be involved in the development of human hepatocellular carcinoma (HCC). So far, most attempts at elucidating HBx function have been undertaken at the genetic level, reflecting the difficulties in detecting the very low amounts of the protein in infected livers. Consequently, the questions of intracellular localization and posttranslational modification have not yet been completely answered. We therefore constructed recombinant baculoviruses that allowed expression of HBx and the hexa histidine HBx fusion protein HBxHis in insect cells. Cell fractionation experiments revealed that only a minor part of HBx is detectable in a soluble form in the cytosolic fraction, whereas most of the protein forms intracellular aggregates. These results could be confirmed by confocal laser immunofluorescence. The fusion of a hexa-histidine tag to the amino terminus of HBx allowed a rapid one-step purification by metal chelate affinity chromatography. The detailed analysis of purified HBxHis using electrospray ionization mass spectrometry uncovered two major components: the unmodified, monomeric, fully oxidized form with five intramolecular disulfide bridges, and its N-acetylated modification. Additionally, two minor peaks with mass differences of delta m = +80 da suggested that a small fraction of HBx becomes posttranslationally phosphorylated in insect cells. No further modifications could be observed, indicating that only phosphorylation might play a role in a possible posttranslational regulation of this viral activator. PMID- 9328334 TI - Cytochromes P450 and uridine triphosphate-glucuronosyltransferases: model autoantigens to study drug-induced, virus-induced, and autoimmune liver disease. AB - Enzymes of phase I (cytochromes P450) and phase II (UDP [uridine diphosphate] glucuronosyltransferases) of drug metabolism are targets of autoimmunity in the following chronic liver diseases of different etiology: 1)autoimmune hepatitis (AIH); 2) hepatitis associated with the autoimmune polyendocrine syndrome type 1 (APS-1); 3) virus-induced autoimmunity; and 4) drug-induced hepatitis. AIH is diagnosed by the following: the absence of infection with hepatitis viruses; the presence of a threshold of relevant factors, including circulating autoantibodies, hypergammaglobulinemia, female sex (female/male ratio 4:1), human leukocyte antigen (HLA) B8, DR3, or DR4; and benefit from immunosuppression. Patients with autoimmune hepatitis type 2 (AIH-2) are characterized by antibodies directed against liver and kidney microsomes, by an early onset of autoimmune hepatitis, which is a more aggressive course of the disease, and by a higher prevalence of autoimmunity directed against other organs. The major target of autoimmunity in patients with AIH-2 is cytochrome P450 2D6. Epitope mapping experiments revealed four short linear epitopes on cytochrome P450 2D6, recognized by liver/kidney microsomal autoantibodies type 1 (LKM-1) in patients with AIH-2. In addition, about 10% of the patient sera contain autoantibodies that detect a conformational epitope on UDP-glucuronosyltransferases (UGTs) of family 1. Presently, LKM-1 autoantibodies are used as diagnostic markers for AIH 2. It is unclear whether these autoantibodies have a pathogenetic role. Hepatitis is found in some patients with APS-1. Presumably this also is an autoimmune liver disease. APS-1 patients with hepatitis may develop autoantibodies directed against microsomal P450 enzymes of the liver; however, these autoantibodies do not recognize cytochrome P450 2D6, but they do recognize cytochrome P450 1A2. Autoimmunity in patients with APS-1 usually is directed against several organs simultaneously, and several organ specific autoantibodies may exist. Interestingly, APS-1 patients may produce various anti-cytochrome P450 antibodies. In addition to the hepatic anti-cytochrome P450, 1A2 autoantibodies are directed against steroidogenic cytochromes P450, namely P450 c21, P450 scc, and P450 c17. These autoantibodies correlate with adrenal and ovarian failure and often these steroidal cell autoantibodies precede the manifestation of adrenal or ovarian dysfunction. Whether anti-P450 1A2 autoantibodies have a similar predictive value is not yet known. LKM autoantibodies are further found in association with chronic hepatitis C and D. In chronic hepatitis C, the major target of LKM autoantibodies is cytochrome P450 2D6. Predominantly, conformational epitopes are recognized by LKM-1 sera of patients with chronic hepatitis C. In 13% of patients with chronic hepatitis D, LKM-3 autoantibody is detectable. The target proteins are UGTs of family 1 and in a minority of sera UGTs of family 2. The epitopes are conformational. All hepatic diseases discussed earlier have in common that autoimmunity, which is directed against enzymes of drug metabolizing multigene families. Each disease is characterized by a specific pattern of autoantibodies, with apparently little overlap. For example, LKM-1 autoantibodies, which are directed against P450 2D6, seem to overlap between AIH and chronic hepatitis C. However, a close examination of these autoantibodies shows differences between LKM-1 autoantibodies from patients with chronic hepatitis C and with AIH. In AIH, LKM autoantibodies are more homogenous, titers are higher, and major autoepitopes on cytochrome P450 2D6 are small and linear. LKM autoantibodies in viral hepatitis C are more heterogeneous and there are multiple epitopes, many of which are conformational. These differences indicate the different mechanisms that are involved in the generation of autoimmunity. (ABSTRACT TRUNCATED) PMID- 9328335 TI - Retinoids may increase fibrotic potential of TGF-beta: crosstalk between two multi-functional effectors. PMID- 9328336 TI - Matrix degradation in liver: a role in injury and repair. PMID- 9328337 TI - Of plaques and stones: the SR-B1 (scavenger receptor class B, type 1). PMID- 9328338 TI - Chronic hepatitis C: beware the older drinking male: fibrosis progression beckons! PMID- 9328339 TI - Hepatitis C virus genotype 1b and risk of hepatocellular carcinoma. PMID- 9328340 TI - Functional synergy between the transcription factor Sp1 and the estrogen receptor. AB - A GC-rich oligonucleotide containing an estrogen responsive element (ERE) half site from the heat shock protein 27 (Hsp 27) gene promoter (-105 to -84) [ie. GGGCGGG(N)10GGTCA; Sp1(N)10ERE] forms a complex with the Sp1 and estrogen receptor (ER) proteins. Moreover, promoter-reporter constructs containing this sequence (-108 to -84 or -108 to +23) are also estrogen-responsive. Mutation of the ERE half-site in the Hsp 27-derived oligonucleotides did not result in loss of estrogen responsiveness in transient transfection studies, suggesting that estrogen inducibility was mediated through the Sp1-DNA motif. Gel mobility shift assays using 32P-labeled wild type and ERE mutant Sp1(N)10ERE and consensus Sp1 oligonucleotides showed that Sp1 protein formed a DNA-protein complex with all three nucleotides, and the intensities of retarded bands were enhanced by coincubation with wild type ER and 11C-ER, which does not contain the DNA-binding domain. ER mutants in which N-terminal (19C-ER) and C-terminal (15C-ER) regions were deleted did not enhance Sp1-DNA binding or hormone-induced transactivation of GC-rich promoter-reporter constructs in ER-negative MDA-MB-231 cells, whereas both wild type and 11C-ER restored inducibility. Immunoprecipitation studies also confirmed that the Sp1 and ER proteins physically interact. The interaction of the Sp1 and ER proteins and the resulting enhanced Sp1-DNA binding is observed in the presence or absence of estrogen (hormone-independent), whereas transactivation of promoter-reporter constructs is estrogen-dependent. Thus, the results illustrate a new estrogen-dependent transactivation pathway that involves ER-protein interactions and is ERE-independent. PMID- 9328341 TI - Interactions of estrogen- and thyroid hormone receptors on a progesterone receptor estrogen response element (ERE) sequence: a comparison with the vitellogenin A2 consensus ERE. AB - The identification of hormone response elements in the promoter regions of hormonally regulated genes has revealed a striking similarity between the half site of the estrogen-response element (ERE) and a consensus sequence constituting the thyroid hormone-response element. Because of the potential for thyroid hormone (T3) to affect estrogen (E)- and progesterone-dependent female reproductive behavior via EREs, we have begun to investigate the activity of an ERE identified in the progesterone receptor (PR) proximal promoter and its interactions with the estrogen receptor (ER) and thyroid hormone receptors (TR). In addition, we have compared ER and TR interactions on the PR ERE construct with that of the vitellogenin A2 (vit A2) consensus ERE. Electrophoretic mobility shift assays demonstrated that TR binds to the PR ERE as well as to the consensus ERE sequence in vitro. Further, these two EREs were differentially regulated by T3 in the presence of TR. T3 in the presence of TR alpha increased transcription from a PR ERE construct approximately 5-fold and had no inhibitory effect on E induction. Similarly, T3 also activated a beta-galactosidase reporter construct containing PR promoter sequences spanning -1400 to +700. In addition, the TR isoforms beta1 and beta2 also stimulated transcription from the PR ERE construct by 5- to 6-fold. A TR alpha mutant lacking the ability to bind AGGTCA sequences in vitro failed to activate transcription from the PR ERE construct, demonstrating dependence on DNA binding. In contrast to its actions on the PR ERE construct, TR alpha did not activate transcription from the vit A2 consensus ERE but rather attenuated E-mediated transcriptional activation. Attenuation from the vit A2 consensus ERE is not necessarily dependent on DNA binding as the TR alpha DNA binding mutant was still able to inhibit E-dependent transactivation. In contrast to TR alpha, the isoforms TRbeta1 and TRbeta2 failed to inhibit E induced activation from the vit A2 consensus ERE. These results demonstrate that the PR ERE construct differs from the vit A2 consensus ERE in its ability to respond to TRs and that divergent pathways exist for activation and inhibition by TR. Since ERs, PRs, and TRs are all present in hypothalamic neurons, these findings may be significant for endocrine integration, which is important for reproductive behavior. PMID- 9328342 TI - Biphasic regulation of breast cancer cell growth by progesterone: role of the cyclin-dependent kinase inhibitors, p21 and p27(Kip1). AB - Depending on the tissue, progesterone is classified as a proliferative or a differentiative hormone. To explain this paradox, and to simplify analysis of its effects, we used a breast cancer cell line (T47D-YB) that constitutively expresses the B isoform of progesterone receptors. These cells are resistant to the proliferative effects of epidermal growth factor (EGF). Progesterone treatment accelerates T47D-YB cells through the first mitotic cell cycle, but arrests them in late G1 of the second cycle. This arrest is accompanied by decreased levels of cyclins D1, D3, and E, disappearance of cyclins A and B, and sequential induction of the cyclin-dependent kinase (cdk) inhibitors p21 and p27(Kip1). The retinoblastoma protein is hypophosphorylated and extensively down regulated. The activity of the cell cycle-dependent protein kinase, cdk2, is regulated biphasically by progesterone: it increases initially, then decreases. This is consistent with the biphasic proliferative increase followed by arrest produced by one pulse of progesterone. A second treatment with progesterone cannot restart proliferation despite adequate levels of transcriptionally competent PR. Instead, a second progesterone dose delays the fall of p21 and enhances the rise of p27(Kip1), thereby intensifying the progesterone resistance in an autoinhibitory loop. However, during the progesterone-induced arrest, the cell cycling machinery is poised to restart. The first dose of progesterone increases the levels of EGF receptors and transiently sensitizes the cells to the proliferative effects of EGF. We conclude that progesterone is neither inherently proliferative nor antiproliferative, but that it is capable of stimulating or inhibiting cell growth depending on whether treatment is transient or continuous. We also suggest that the G1 arrest after progesterone treatment is accompanied by cellular changes that permit other, possibly tissue-specific, factors to influence the final proliferative or differentiative state. PMID- 9328343 TI - Analysis of the role of E2A-encoded proteins in insulin gene transcription. AB - Pancreatic beta-cell type-specific transcription of the insulin gene is mediated, in part, by factors in the basic helix-loop-helix (bHLH) family that act on a site within the insulin enhancer, termed the E1-box. Expression from this element is regulated by a heteromeric protein complex containing ubiquitous (i.e. the E2A and HEB-encoded proteins) and islet-enriched members of the bHLH family. Recent studies indicate that the E2A- and HEB-encoded proteins contain a transactivation domain, termed AD2, that functions more efficiently in transfected beta-cell lines. In the present report, we extend this observation by demonstrating that expression of full-length E2A proteins (E47, E12, and E2/5) activates insulin E element-directed transcription in a beta-cell line-selective manner. Stimulation required functional interactions with other key insulin gene transcription factors, including its islet bHLH partner as well as those that act on the RIPE3b1 and RIPE3a2 elements of the insulin gene enhancer. The conserved AD2 domain in the E2A proteins was essential in this process. The effect of the E2A- and HEB-encoded proteins on insulin gene expression was also analyzed in mice lacking a functional E2A or HEB gene. There was no apparent difference in insulin production between wild type, heterozygote, and homozygous mutant E2A or HEB mice. These results suggest that neither the E2A- or HEB-encoded proteins are essential for insulin transcription and that one factor can substitute for the other to impart normal insulin E1 activator function in mutant animals. PMID- 9328344 TI - Differential regulation of mitogen-activated protein/ERK kinase (MEK)1 and MEK2 and activation by a Ras-independent mechanism. AB - Mitogen-activated protein (MAP)/ERK kinase (MEK)1 and MEK2 are the upstream activators of the MAP kinases, ERK1 and ERK2. MEK1 and MEK2 are approximately 85% identical in sequence but have unique inserts in their C-terminal domains. MEK isoform-specific antibodies were used to examine expression and regulation of each enzyme. MEK1 and MEK2 were expressed in approximately equal amounts in several cell lines; in some, MEK1 was present in slight excess. Activation of tyrosine kinase-containing receptors, heterotrimeric G proteins, and protein kinase C enhanced the activities of both MEK isoforms in 293 and PC12 cells. AIF4 stimulated both MEK1 and MEK2 in PC12 cells expressing a dominant interfering Ras mutant that prevents nerve growth factor-dependent activation of the cascade. Carbachol also stimulated the pathway in these cells. Thus, in addition to their ability to activate Ras/Raf and the downstream ERK pathway, heterotrimeric G proteins also appear to trigger a Ras-independent mechanism to regulate this kinase cascade. In U373, Chinese hamster ovary (CHO), and INS-1 cells, MEK1 was activated by regulators of ERKs, while MEK2 was not. These data suggest that, like the MAP kinases ERK1 and ERK2, in some cell settings the two similar MEK isoforms are differentially regulated. PMID- 9328345 TI - The activation function-2 hexamer of steroidogenic factor-1 is required, but not sufficient for potentiation by SRC-1. AB - The orphan receptor steroidogenic factor 1 (SF-1) plays a central role in development and differentiation of the adrenal gland and gonads. It also regulates the expression of several pivotal steroidogenic enzymes and other proteins that are essential for reproductive function. Its mechanism of target gene activation that directs these intricate processes has not been previously established. We demonstrate here that the activation function-2 (AF-2) activation hexamer (AF-2-AH) of SF-1, located within its carboxy-terminal region, is required for reporter gene activation by SF-1, as well as for SF-1-mediated induction of a steroidogenic phenotype in embryonic stem cells. We further demonstrate that SF-1's AF-2-AH is not sufficient for gene activation, requiring an additional, proximally located domain of SF-1, positioned between residues 187 245. Correspondingly, we show that the coactivator SRC-1 potentiates the activity of SF-1 and that the interaction between SF-1 and SRC-1 requires both AF-2-AH and the proximal activation domain. We conclude that SF-1 harbors at least two activation domains within its carboxy terminus and that both are required for its transcriptional activation function and for direct interaction with SRC-1. It is likely that SRC-1 plays a key role in gene regulation by SF-1. PMID- 9328346 TI - Colony-stimulating factor-1 plays a major role in the development of reproductive function in male mice. AB - Colony-stimulating factor-1 (CSF-1) is the principal regulator of cells of the mononuclear phagocytic lineage that includes monocytes, tissue macrophages, microglia, and osteoclasts. Macrophages are found throughout the reproductive tract of both males and females and have been proposed to act as regulators of fertility at several levels. Mice homozygous for the osteopetrosis mutation (csfm[op]) lack CSF-1 and, consequently, have depleted macrophage numbers. Further analysis has revealed that male csfm(op)/csfm(op) mice have reduced mating ability, low sperm numbers, and 90% lower serum testosterone levels. The present studies show that this low serum testosterone is due to reduced testicular Leydig cell steroidogenesis associated with severe ultrastructural abnormalities characterized by disrupted intracellular membrane structures. In addition, the Leydig cells from csfm(op)/ csfm(op) males have diminished amounts of the steroidogenic enzyme proteins P450 side chain cleavage, 3beta hydroxysteroid dehydrogenase, and P450 17alpha-hydroxylase-lyase, with associated reductions in the activity of all these steroidogenic enzymes, as well as in 17beta-hydroxysteroid dehydrogenase. The CSF-1-deficient males also have reduced serum LH and disruption of the normal testosterone negative feedback response of the hypothalamus, as demonstrated by the failure to increase LH secretion in castrated males and their lack of response to exogenous testosterone. However, these males are responsive to GnRH and LH treatment. These studies have identified a novel role for CSF-1 in the development and/or regulation of the male hypothalamic-pituitary-gonadal axis. PMID- 9328347 TI - Silencing of the gene for the alpha-subunit of human chorionic gonadotropin by the embryonic transcription factor Oct-3/4. AB - CG is required for maintenance of the corpus luteum during pregnancy in higher primates. As CG is a heterodimeric molecule, some form of coordinated control must be maintained over the transcription of its two subunit genes. We recently found that expression of human CG beta-subunit (hCGbeta) in JAr human choriocarcinoma cells was almost completely silenced by the embryonic transcription factor Oct-3/4, which bound to a unique ACAATAATCA octameric sequence in the hCGbeta gene promoter. Here we report that Oct-3/4 is also a potent inhibitor of hCG alpha-subunit (hCGalpha) expression in JAr cells. Oct-3/4 reduced human GH reporter expression from the -170 hCGalpha promoter in either the presence or absence of cAMP by about 70% in transient cotransfection assays, but had no effect on expression from either the -148 hCGalpha or the -99 hCGalpha promoter. Unexpectedly, no Oct-3/ 4-binding site was identified within the -170 to -148 region of the hCGalpha promoter, although one was found around position 115 by both methylation interference footprinting and electrophoretic mobility shift assays. Site-directed mutagenesis of this binding site destroyed the affinity of the promoter for Oct-3/4, but did not affect repression of the promoter. Therefore, inhibition of hCGalpha gene transcription by Oct-3/4 appears not to involve direct binding of this factor to the site responsible for silencing. When stably transfected into JAr cells, Oct-3/4 reduced the amounts of both endogenous hCGalpha mRNA and protein by 70-80%. Oct-3/4 is therefore capable of silencing both hCGalpha and hCGbeta gene expression. We suggest that as the trophoblast begins to form, reduction of Oct-3/4 expression permits the coordinated onset of transcription from the hCGalpha and hCGbeta genes. PMID- 9328348 TI - Derivation of functional antagonists using N-terminal extracellular domain of gonadotropin and thyrotropin receptors. AB - Receptors for the glycoprotein hormones, LH/CG, FSH, and TSH, are a unique subclass of the seven-transmembrane, G protein-coupled proteins with a large N terminal extracellular (ecto-) domain. Although ecto-domains of gonadotropin receptors confer ligand binding, expression of soluble binding proteins has been difficult. We fused the ecto-domains of LH or FSH receptors to the single transmembrane domain of CD8 and found that hybrid proteins anchored on the cell surface retained high-affinity ligand binding. Inclusion of a junctional thrombin cleavage site in the hybrids allowed generation of soluble receptor fragments that interfered with gonadotropin binding to their receptors and blocked cAMP production stimulated by gonadotropins. Cross-linking analyses confirmed the formation of high molecular weight complexes between receptor ecto-domains and their specific ligands. A similar approach also generated a soluble TSH receptor fragment capable of blocking TSH-induced signal transduction. When administered to rats, the soluble FSH receptor fragment retarded testis growth and induced testis cell apoptosis. These findings demonstrate the feasibility of generating ligand-binding regions of glycoprotein hormone receptors to selectively neutralize actions of gonadotropins and TSH, thus allowing future design of novel contraceptives and management of different gonadal and thyroid dysfunction. The present study represents the first successful derivation of soluble, ligand binding domains from glycoprotein hormone receptors as functional antagonists. Similar approaches could allow generation of ecto-domains of related receptors to neutralize actions of ligands or receptor antibodies and to facilitate structural functional analysis. PMID- 9328349 TI - Multiple characteristics of a pentameric regulatory array endow the human alpha subunit glycoprotein hormone promoter with trophoblast specificity and maximal activity. AB - Trophoblast-specific expression of the human alpha-subunit glycoprotein hormone gene requires a tightly linked array of five different regulatory elements [trophoblast-specific element (TSE), alpha-activating element (alphaACT), a tandem cAMP response element (CRE), junctional regulatory element (JRE), and a CCAAT box]. We examined their contextual contributions to trophoblast-specific expression by using transfection assays to evaluate activity of systematic block replacement mutations made within the 1500-bp 5'-flanking region of the human alpha-subunit gene. While all five elements were required for full activity, only the TSE and JRE displayed trophoblast specificity. Interestingly, the TSE-binding protein has limited tissue distribution whereas a JRE-binding protein appears trophoblast specific. Likewise, replacement studies with an AP-1 element that binds heterodimers of jun and fos indicated that this element was incapable of compensating for either the tandem CRE or JRE. This preference for both CRE- and JRE-binding proteins provides another avenue for configuring an alpha-subunit promoter with trophoblast specificity. Additional analysis with a cAMP response element binding protein (CREB)-Gal4 fusion protein further underscored the importance of CREB as well as suggested that transcriptional contributions come from both the DNA-binding domain and transactivation domain of this protein. We also examined the interactive nature of the pentameric array by placing a 15-bp random sequence between each element. Remarkably, only the insertion 3' of the CCAAT box diminished promoter activity. This suggested the absence of direct interactions between the transcriptional factors that bind each element in the array. It also suggested that the CCAAT box is position-dependent relative to the TATA box. This position dependence appeared cell-specific, as it was not manifest in a gonadotrope cell line (alphaT3-1 cells). Thus, the CCAAT box also has tissue specific characteristics that assist in targeting expression of the alpha-subunit gene to trophoblasts. Together, these data suggest that multiple characteristics of a complex pentameric array of regulatory elements endow the alpha-subunit promoter with trophoblast specificity and maximal activity. PMID- 9328350 TI - Stage-specific expression of Dlx-5 during osteoblast differentiation: involvement in regulation of osteocalcin gene expression. AB - Two homeotic genes, Dlx and Msx, appear to regulate development of mineralized tissues, including bone, cartilage, and tooth. Expression of Msx-1 and Msx-2 has been studied during development of the osteoblast phenotype, but the role of Dlx in this context and in the regulation of bone-expressed genes is unknown. We used targeted differential display to isolate homeotic genes of the Dlx family that are expressed at defined stages of osteoblast differentiation. These studies were carried out with fetal rat calvarial cells that produce bone-like tissue in vitro. We observed a mineralization stage-specific mRNA and cloned the corresponding cDNA, which represents the rat homolog of Dlx-5. Northern blot analysis and competitive RT-PCR demonstrated that Dlx-5 and the bone-specific osteocalcin genes exhibit similar up-regulated expression during the mineralization period of osteoblast differentiation. This expression pattern differs from that of Msx-2, which is found predominantly in proliferating osteoblasts. Several approaches were pursued to determine functional consequences of Dlx-5 expression on osteocalcin transcription. Constitutive expression of Dlx 5 in ROS 17/2.8 cells decreased osteocalcin promoter activity in transient assays, and conditional expression of Dlx-5 in stable cell lines reduced endogenous mRNA levels. Consistent with this finding, antisense inhibition of Dlx 5 increased osteocalcin gene transcription. Osteocalcin promoter deletion analysis and binding of the in vitro translation product of Dlx-5 demonstrated that repressor activity was targeted to a single homeodomain-binding site, located in OC-Box I (-99 to -76). These findings demonstrate that Dlx-5 represses osteocalcin gene transcription. However, the coupling of increased Dlx-5 expression with progression of osteoblast differentiation suggests an important role in promoting expression of the mature bone cell phenotype. PMID- 9328351 TI - Human and murine osteocalcin gene expression: conserved tissue restricted expression and divergent responses to 1,25-dihydroxyvitamin D3 in vivo. AB - Human and murine osteocalcin genes demonstrate similar cell-specific expression patterns despite significant differences in gene locus organization and sequence variations in cis-acting regulatory elements. To investigate whether differences in these regulatory regions result in an altered response to 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3] in vivo, we compared the response of the endogenous mouse osteocalcin gene to a bacterial reporter gene directed by flanking regions of the human osteocalcin gene in transgenic mice. Transgene expression colocalized with endogenous osteocalcin expression in serial sections, being detected in osteoblasts, osteocytes and hypertrophic chondrocytes. In calvarial cell culture lysates from transgenic and nontransgenic mice, the endogenous mouse osteocalcin gene did not respond to 1,25-(OH)2D3 treatment. Despite this, transgene activity was significantly increased in the same cells. Similarly, Northern blots of total cellular RNA and in situ hybridization studies of transgenic animals demonstrated a maximal increase in transgene expression at 6 h after 1,25-(OH)2D3 injection (23.6+/-3.6-fold) with a return to levels equivalent to uninjected animals by 24 h (1.2+/-0.1-fold). This increase in transgene expression was also observed at 6 h after 1,25-(OH)2D3 treatment in animals on a low calcium diet (25.2+/-7.7-fold) as well as in transgenic mice fed a vitamin D-deficient diet containing strontium chloride to block endogenous 1,25 (OH)2D3 production (7.5+/-0.9-fold). In contrast to the increased transgene expression levels, neither endogenous mouse osteocalcin mRNA levels nor serum osteocalcin levels were significantly altered after 1,25-(OH)2D3 injection in transgenic or nontransgenic mice, regardless of dietary manipulations, supporting evidence for different mechanisms regulating the response of human and mouse osteocalcin genes to 1,25-(OH)2D3. Although the cis- and trans-acting mechanisms directing cell-specific gene expression appear to be conserved in the mouse and human osteocalcin genes, responsiveness to 1,25-(OH)2D3 is not. The mouse osteocalcin genes do not respond to 1,25-(OH)2D3 treatment, but the human osteocalcin-directed transgene is markedly upregulated under the same conditions and in the same cells. The divergent responses of these homologous genes to 1,25 (OH)2D3 are therefore likely to be due to differences in mouse and human osteocalcin-regulatory sequences rather than to variation in the complement of trans-acting factors present in mouse osteoblastic cells. Increased understanding of these murine-human differences in osteocalcin regulation may shed light on the function of osteocalcin and its regulation by vitamin D in bone physiology. PMID- 9328352 TI - Somatostatin receptor subtype 2 knockout mice are refractory to growth hormone negative feedback on arcuate neurons. AB - The pulsatile nature of GH release is apparently regulated by alternating sequential changes in two hypothalamic hormones, GH releasing hormone (GHRH) and somatostatin. Entrainment of this pulsatility appears to involve GH-mediated negative feedback. Recently a new receptor involved in GH release was cloned. Activation of this receptor by GH-releasing peptides and MK-0677 initiates and amplifies GH pulsatility and is associated with increased Fos immunoreactivity and electrical activity in GHRH containing arcuate neurons. We show that pretreating mice with GH blocks activation of these neurons by MK-0677. Similarly, octreotide inhibited the action of MK-0677. To determine whether this GH-mediated negative feedback on GHRH neurons was direct, or by GH stimulation of somatostatin release from periventricular neurons, we selectively inactivated the gene for one of the five specific somatostatin receptor subtypes (subtype 2). In the knockout mice, both GH and octreotide failed to inhibit MK-0677 activation of arcuate neurons. GH did, however, increase Fos immunoreactivity in the periventricular nucleus, consistent with GH stimulation of somatostatin release from periventricular neurons. Thus, GH-mediated negative feedback involves signaling between periventricular and arcuate neurons with the signal being transduced specifically through somatostatin subtype 2 receptors. PMID- 9328353 TI - A novel mutation adjacent to the switch III domain of G(S alpha) in a patient with pseudohypoparathyroidism. AB - A novel G(S alpha) mutation encoding the substitution of arginine for serine 250 (G[S alpha] S250R) was identified in a patient with pseudohypoparathyroidism type Ia. Both G(S) activity and G(S alpha) expression were decreased by about 50% in erythrocyte membranes from the affected patient. The cDNA of this G(S alpha) mutant, as well as one in which the S250 residue is deleted (G[S alpha] deltaS250), was generated, and the biochemical properties of the products of in vitro transcription/translation were examined. Both mutants had a sedimentation coefficient similar to that of wild type G(S alpha) (approximately 3.7S) when kept at 0 C after synthesis. However when maintained for 1-2 h at 30-37 C, both mutants aggregated to a material sedimenting at approximately 6.3S or greater (G[S alpha]-S250R to a greater extent than G(S alpha]-deltaS250), while wild type G(S alpha) sedimented at approximately 3.7S, suggesting that the mutants were thermolabile. Incubation in the presence of high doses of guanine nucleotide partially prevented heat denaturation of G(S alpha) deltaS250 but had no protective effect on G(S alpha-S250R. Sucrose density gradient centrifugation at 0 C in the presence and absence of beta gamma-dimers demonstrated that, in contrast to wild type G(S alpha) neither mutant could interact with beta gamma. Trypsin protection assays revealed no protection of G(S alpha)-S250R by GTPgammaS or AIF4- at any temperature. GTPgammaS conferred modest protection of G(S alpha) deltaS250 (approximately 50% of wild-type G[S alpha]) at 30 C but none at 37 C, while AIF4- conferred slight protection at 20 C but none at 30 C or above. Consistent with this result, G(S alpha)-deltaS250 was able to stimulate adenylyl cyclase at 30 C when reconstituted with cyc- membranes in the presence of GTPgammaS but not in the presence of AIF4-. G(S alpha)-S250R showed no ability to stimulate adenylyl cyclase in the presence of either agent. Stable transfection of mutant and wild-type G(S alpha) into cyc- S49 lymphoma cells revealed that the majority of wild type G(S alpha) localized to membranes, while little or no membrane localization occurred for either mutant. Modeling of G(S alpha) based upon the crystal structure of G(t alpha) or G(i alpha) suggests that Ser250 interacts with several residues within and around the conserved NKXD motif, which directly interacts with the guanine ring of bound GDP or GTP. It is therefore possible that substitution or deletion of this residue may alter guanine nucleotide binding, which could lead to thermolability and impaired function. PMID- 9328354 TI - Purification, molecular cloning, and functional expression of the human nicodinamide-adenine dinucleotide phosphate-regulated thyroid hormone-binding protein. AB - The kidney and several other thyroid hormone-responsive tissues contain a NADP regulated thyroid hormone (TH)-binding protein (THBP), with an apparent molecular mass of 36 kDa on SDS-PAGE, responsible for most of the intracellular high affinity T3 and T4 binding. THBP was purified to homogeneity from human kidney cytosol and used to generate proteolytic peptides. Microsequencing of four peptides revealed identity to amino acid sequences deduced from a human cDNA homolog to a cDNA encoding kangaroo mu-crystallin. This protein is a major structural kangaroo lens protein with no known function in other species. A full sized cDNA (TH5.9) was isolated by 5'- and 3'-rapid amplification of cDNA ends using a human brain cDNA library and gene-specific PCR primers, confirming identity to the previously cloned human cDNA. The TH5.9 cDNA encodes a 314 residue protein (theoretical mol wt = 33,775) with significant homologies (40 to 60%) with two bacterial enzymes: lysine cyclodeaminase and ornithine cyclodeaminase. The TH5.9 cDNA was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. Purified GST fusion protein, but not GST, bound T3 specifically with high affinity [dissociation constant (Kd) = 0.5 nM] in the presence of NADPH, and was labeled by UV-driven cross-linking of underivatized [(125)I]T3. T3 binding and photoaffinity labeling of GST fusion protein were activated by NADPH [activation constant (K[act]) = 10(-8) M], but not by NADH. The expressed protein displays the appropriate binding properties, indicating that TH5.9 cDNA encodes the NADP-regulated THBP characterized in human tissues. PMID- 9328356 TI - Thyroid transcription factor-1 activates the promoter activity of rat thyroid Na+/I- symporter gene. AB - We have cloned 15 kbp of rat thyroid Na+/I- symporter gene from liver genomic library, which contains 6 kbp upstream sequence from the translation initiation site. Southern blot analysis of the genomic DNA from the liver has revealed that thyroid Na+/I- symporter gene is the single gene in the rat. To study the tissue selective expression mechanism of the gene, we at first determined the transcriptional start site of the gene. Results of a rapid amplification of cDNA end procedure as well as that of primer extension analysis indicated that the transcriptional start sites clustered between -96, -95, and -93 bp of the gene (A in ATG is designated as +1). Chimeras containing 1.9 kbp (-1967 to -46 bp) of the 5'-flanking sequence of the Na+/I- symporter gene and luciferase gene expressed significant enzyme activity when transfected into a rat thyroid cell line, FRTL 5, but little activity was observed in BRL-3A rat liver cells. Deletion analysis of the constructs indicated that a minimal region, exhibiting promoter activity and cell specificity, is located between -291 and -134 bp of the gene. Deoxyribonuclease I footprinting shows that nuclear extracts from FRTL-5, but not BRL-3A, cells protect a region between -245 and -230 bp. Electrophoretic mobility shift assays have demonstrated that nuclear extracts from FRTL-5 cells formed a specific DNA-protein complex with an oligonucleotide probe corresponding to -250 to -211 bp of the gene, but that from BRL-3A cells did not, suggesting that thyrocyte-selective nuclear factors bind to the region. When the nuclear extracts from FRTL-5 cells were preincubated with antibody against thyroid transcription factor-1 (TTF-1), homeodomain containing nuclear protein, formation of the complex was abolished and the band was supershifted. We also found that the probe formed a DNA-protein complex with the recombinant TTF-1 homeodomain, and mutations of the binding site eliminated factor binding. When pRc/CMV-TTF-1 was cotransfected with the minimal promoter fragment of thyroid Na+/I- symporter gene into FRT cells, which express no TTF-1, it caused a significant increase in the transcriptional activity of the reporter construct, but not of the construct having mutated TTF-1-binding element. These results suggest that TTF-1 confers the cell-selective expression of Na+/I- symporter gene in thyrocytes. PMID- 9328355 TI - Transcriptional activation and repression by RORalpha, an orphan nuclear receptor required for cerebellar development. AB - Mutation of the orphan nuclear receptor RORalpha results in a severe impairment of cerebellar development by unknown mechanisms. We have found that RORalpha activates transcription from only a subset of sites to which it binds strongly as a monomer. RORalpha also selectively binds as a homodimer to a direct repeat of this monomer site with a 2-bp spacing between the AGGTCA sequences (Rev-DR2 site) and is a much more potent transcriptional activator on this site than on monomer sites or other direct repeats. To better understand the transcriptional regulatory functions of RORalpha, we fused its C terminus to a heterologous DNA binding domain. Mutational analysis revealed that RORalpha contains both transcriptional activation and transcriptional repression domains, with the repression domain being more active in some cell types. The abilities of RORalpha polypeptides to repress transcription correlate with their abilities to interact with the nuclear receptor corepressors N-CoR and SMRT in vitro. However, the AF2 region of RORalpha inhibits corepressor interaction on DNA, consistent with the lack of repression by the full-length receptor. Thus, transcriptional regulation by RORalpha is complex and likely to be regulated in a cell type- and target gene specific manner. PMID- 9328357 TI - Use of diuretics in chronic renal failure. PMID- 9328358 TI - A critical evaluation of ultrasound measurement of inferior vena cava diameter in assessing dry weight in normotensive and hypertensive hemodialysis patients. AB - The utility of measurement of the inferior vena cava diameter (IVCD) with ultrasound for the assessment of fluid status and posthemodialysis dry weight was studied in 35 hemodialysis (HD) patients, 17 with and 18 without hypertension. In 17 patients (group A), IVCD was measured before and 35 to 40 minutes after HD, pre-HD blood volume (BV) was measured with radiolabeled albumin and post-HD BV was calculated from the change in hematocrit. In 18 patients (group B), IVCD was measured repeatedly during HD and 2 hours after HD. Changes in BV were recorded by monitoring of the hematocrit "on line." Body weight, blood pressure (BP), BV, and IVCD decreased in the entire population. In group A, BV was significantly larger in the hypertensive patients than in the normotensive patients, and it was correlated with the mean BP before and after HD. In the whole population, IVCD was larger in the hypertensive than in the normotensive patients before and after HD. These results confirm that extracellular fluid overload plays an important role in the pathogenesis of dialysis-associated hypertension. In group B, BV and IVCD decreased in parallel during HD and increased during 2 hours after HD due to refilling of the intravascular space, indicating that changes in IVCD reflect changes in BV. In 8 patients studied twice, IVCD increased much more after a 3 hour HD session than after a 6-hour session. At the end of HD, several patients had IVCD below the reference range but IVCD increased during the following 1 to 2 hours, in some patients to values above the reference range. IVCD measured at the end or shortly after HD may therefore be misleading in assessing dry weight. PMID- 9328359 TI - Effects of hypervolemia on interdialytic hemodynamics and blood pressure control in hemodialysis patients. AB - The influence of hypervolemia on hemodynamics and interdialytic blood pressure, as well as in relation to vascular compliance, was investigated in 10 hemodialysis patients who were not receiving vasoactive medication. All subjects were studied during a relative normovolemic interdialytic period (from 1 kg below dry weight postdialytic until dry weight predialytic) and a hypervolemic interdialytic period (from 1 kg above dry weight postdialytic until 3 kg above dry weight predialytic). Interdialytic blood pressure was measured with an ambulatory blood pressure monitor. Cardiac output was echographically measured and total peripheral resistance calculated postdialytic, mid-interdialytic, and predialytic. At the same time, a blood sample was drawn for analyzing vasoactive hormones, sodium, and hematocrit. In all patients, ideal dry weight was estimated by echography of the caval vein. Arterial and venous compliance were measured with an ultrasound vessel wall movement detector system and a strain-gauge plethysmograph. After fluid load, an increase in intravascular volume, an increase in caval vein diameter and cardiac output, and a decrease in peripheral resistance was observed. No significant influence of a 3-L fluid load was found on interdialytic blood pressure course (153+/-24 mm Hg/90+/-19 mm Hg in the hypervolemic period and 146+/-27 mm Hg/89+/-22 mm Hg in the normovolemic period). Sodium and osmolality were similar in the hypervolemic and normovolemic interdialytic periods. After fluid load, a decrease in arginine vasopressin and angiotensin II was observed, which probably contributed to the decreased systemic vascular resistance. Catecholamines were not influenced by fluid load, but increased during the interdialytic period, suggesting accumulation after dialysis. Three of the 10 patients had higher systolic but not diastolic blood pressures after fluid load (159+/-13 mm Hg/81+/-22 mm Hg in the hypervolemic period and 135+/-16 mm Hg/81+/-22 mm Hg in the normovolemic period). No correlation could be found between arterial or venous compliance and blood pressure changes. We concluded that a 3-L interdialytic fluid load does not result in higher blood pressure in most hemodialysis patients. PMID- 9328360 TI - Hemodialysis access blood flow rates can be measured by a differential conductivity technique and are predictive of access clotting. AB - Blood flow in peripheral arteriovenous fistulae and grafts as used for hemodialysis access can be derived from measurements of the amount of access recirculation induced by reversing the dialysis blood lines and a knowledge of the dialyzer blood flow rate. The Hemodynamic Monitor (HDM; GAMBRO Healthcare, Lakewood, CO) is a device that uses magnetic principles to accurately and precisely measure access recirculation during hemodialysis. The measurement is based on differential conductivity between arterial and venous blood flows in the dialysis blood tubing sets following the injection of hypertonic saline into the venous line as a conductivity tracer. Clinical studies were performed on 41 patients from two centers who had arteriovenous fistulae (25 patients) or Goretex grafts (16 patients; W.L. Gore & Associates, Flagstaff, AZ); each patient was studied on two successive dialysis days under variable conditions of dialyzer blood flow, and multiple measurements were made according to a standard protocol. The protocol involved temporarily reversing the arterial and venous lines, then performing an HDM recirculation test and recording the result along with the dialyzer blood flow rate as per the machine blood pump setting. The access blood flow rates measured 1,125+/-581 mL/min (mean+/-SD) on day 1 and 1,140+/-680 mL/ min on day 2 (P > 0.05 [NS]), with an absolute range of 221 to 3,118 mL/min. These flow rates are similar to those measured by other techniques. There was an excellent correlation between access blood flow rates measured in individual patients on days 1 and 2, even in a subset of 13 patients who had the dialyzer blood flow rates altered by > or =100 mL/min, suggesting the independence of access from dialyzer blood flow rates. Analysis of repeated measurements of access blood flow under identical conditions showed a characteristic standard deviation from the mean across the patient population of 7.89%, indicating that the HDM results are repeatable in clinical application. The influence of the measured access blood flow on the outcome of that access was determined after an 8-month follow-up period. Of the 41 accesses, nine were lost to clotting; seven of 14 that had initial blood flow rates less than 750 mL/min clotted, while only two of 27 with flow rates greater than 750 mL/min subsequently clotted (P = 0.005). The data show that the HDM can provide clinically important information on access blood flow. PMID- 9328361 TI - Cytokines clearance during venovenous hemofiltration in the trauma patient. AB - The objective of the study was to investigate whether continuous venovenous hemofiltration (CVVH) would facilitate removal of substantial amounts of tumor necrosis factor (TNF) and interleukin-6 (IL-6) from the circulation in traumatized critically ill patients with multiple organ dysfunction syndrome. The study design was a prospective, nonblind, randomized controlled trial that was set in the trauma intensive care unit of a tertiary university referral hospital. Thirty consecutive critically ill, mechanically ventilated trauma patients with multiple organ dysfunction syndrome (without renal failure) were included in the study. Patients were randomized to either CVVH or conventional treatment. Blood and ultrafiltrate samples were collected from each patient before the initiation of CVVH and after 24, 72, and 168 hours of therapy. In the control group, blood samples were collected during the same periods. In the 30 patients studied, 15 had hemofiltration and 15 did not. Both groups were similar with regard to age (36+/-18 years v 36+/-14 years) and severity scores (injury severity score, 32+/ 16 v 30+/-11; APACHE II score, 22+/-7 v 21+/-6; Goris score, 5.2+/-1.7 v 5.2+/ 1.8). Before CVVH, TNF and IL-6 could be detected in the serum of all patients. The mean concentration of TNF was 17+/-22 pg/mL in patients and 22+/-20 pg/mL in control subjects (P = NS). The mean concentration of IL-6 was 2,153+/-2,824 pg/mL in patients and 1,774+/-1,637 pg/mL in control subjects (P = NS). We found a TNF and IL-6 substantial elimination with CVVH (excretion of TNF [microg/d] at 24, 48, and 168 hours: 112.6+/-161.2, 105.2+/-149.4, and 143.1+/-170.0; excretion of IL-6 [microg/d]: 1,655+/-719, 3,091+/-489, and 2,420+/-366). However, no significant difference was found in serum cytokines concentration between groups during the study: mean serum TNF concentration decreased from the pretreatment level to a mean level of 12+/-9.6 pg/mL in patients and 21+/-27 pg/mL in control subjects. Similar results were found with IL-6 concentration that decreased from the pretreatment level to a mean of 554+/-731 pg/mL in patients and 382 +/-568 pg/mL in control subjects. In conclusion, CVVH is associated with removal of substantial amounts of TNF and IL-6 from the circulation in traumatized critically ill patients, but the profile of these mediators is similar to that of controls, suggesting a nonclinically relevant elimination. Further prospective, randomized, clinical trials are needed to support our results. PMID- 9328362 TI - Low whole blood and erythrocyte levels of glutathione in hemodialysis and peritoneal dialysis patients. AB - Dialysis patients are reported to have impaired antioxidant mechanisms, including those involving glutathione-dependent enzymes. This study used high-performance liquid chromatography assays that directly measure total (oxidized + reduced) glutathione and its precursor cysteine (CYS) to compare the whole blood of hemodialysis (prehemodialysis and posthemodialysis) and peritoneal dialysis patients to that of blood donors with no known kidney disease (n=20 in each group). The levels in erythrocytes were calculated from that data (as nmol/g hemoglobin) because these cells are the major compartment of blood glutathione and their survival may be shortened by oxidant damage. Both dialysis groups had significantly (P=0.0001) higher CYS levels in the plasma compartment than the controls (251 nmol/mL), with prehemodialysis levels (432 nmol/mL) being greater than peritoneal dialysis levels (334 nmol/mL). Hemodialysis acutely lowered CYS levels (215 nmol/mL) below those of controls. Expressed per milliliter whole blood, both dialysis groups had significantly (P=0.0001) lower glutathione levels than controls (1,276 nmol/mL), with prehemodialysis and peritoneal dialysis levels being similar (778 and 912 nmol/mL). Values increased prehemodialysis to posthemodialysis, consistent with hemoconcentration. Expressed per gram hemoglobin, the dialysis groups had significantly (P < 0.015) lower glutathione levels than the controls (8,938 nmol/g hemoglobin), with similar prehemodialysis, posthemodialysis, and peritoneal dialysis values (7,207, 7,315, and 7,915 nmol/g hemoglobin, respectively). In summary, hemodialysis and peritoneal dialysis patients are at increased risk from oxidative stress due to glutathione deficiency in whole blood and erythrocytes. PMID- 9328363 TI - A 6-month study of low-dose recombinant human erythropoietin alone and in combination with androgens for the treatment of anemia in chronic hemodialysis patients. AB - Two previous short-term studies (12 weeks and up to 16 weeks) that used androgens to supplement recombinant human erythropoietin (rHuEPO) for the treatment of the anemia associated with end-stage renal disease showed divergent results. Both studies were limited by their brief duration, since the hematopoietic effect of androgens does not peak until 5 months. Therefore, we conducted a 6-month, prospective, randomized trial comparing low-dose rHuEPO alone and in combination with androgens for the treatment of the anemia of end-stage renal failure. Nineteen anemic chronic hemodialysis patients were randomized into two groups. Group A (n = 10) received 1,500 U rHuEPO intravenously three times a week for 26 weeks. Group B (n = 9) received the same dose of rHuEPO plus nandrolone decanoate 100 mg intramuscularly weekly. Baseline transferrin saturation, serum ferritin, intact serum parathyroid hormone, plasma aluminum, and hematocrit levels were not significantly different between the groups. At study completion, both groups showed a significant increase in mean hematocrit compared with baseline (group A: 24.8% +/- 1.4% to 28.3% +/- 2.8%, P = 0.003; group B: 25.1% +/- 1.5% to 33.2% +/- 4.5%, P = 0.001). The increase in hematocrit in the rHuEPO plus androgen-treated group was statistically greater than in the rHuEPO-alone group (8.2% +/- 4.4% v 3.5% +/- 2.8%; P = 0.012). With the exception of mild discomfort at the injection site, there were no significant side effects from nandrolone. We conclude that the combination of low-dose rHuEPO and nandrolone decanoate is effective treatment for the anemia of end-stage renal failure. PMID- 9328364 TI - Maintaining blood compartment volume in dialyzers reprocessed with peracetic acid maintains Kt/V but not beta2-microglobulin removal. AB - A dialyzer is reused if its blood compartment volume is 80% of its initial value, a condition believed to ensure that the urea clearance remains at 90% of its initial value. This criterion was developed for dialyzers containing low permeability cellulose membranes reprocessed with formaldehyde. We tested the hypothesis that the criterion is also valid for more permeable membranes when dialyzers are reprocessed with peracetic acid/hydrogen peroxide. Kt/V for urea and reduction in beta2-microglobulin concentration were measured for up to 15 uses in dialyzers containing polysulfone or cellulose membranes. Kt/V for urea did not change for either dialyzer provided blood compartment volumes remained 80% of their initial value. The reduction in plasma beta2-microglobulin concentration from predialysis to postdialysis was 30% for the first use of the dialyzer containing polysulfone membranes, but decreased significantly (P = 0.042) following reuse to 12% for the tenth use. For the dialyzers containing cellulose membranes, the reduction in plasma beta2-microglobulin concentration was 18% for the first use and decreased to 12% by the twelfth use; however, this change was not significant. We conclude that removal of urea is maintained during reuse with peracetic acid/hydrogen peroxide provided the blood compartment volume remains 80% of its initial value. However, removal of beta2-microglobulin may not be maintained, even though blood compartment volumes remain at 80% of their initial value. PMID- 9328365 TI - Effect of increasing serum albumin on serum lipoprotein(a) concentration in patients receiving CAPD. AB - Lipoprotein(a) [Lp(a)], an independent risk factor for atherosclerotic cardiovascular disease in the general population, is known to be elevated in patients with renal disease accompanied by hypoalbuminemia such as nephrotic syndrome and end-stage renal disease. In this study, the role of hypoalbuminemia in the elevation of serum Lp(a) was investigated in 20 continuous ambulatory peritoneal dialysis (CAPD) patients with serum albumin below 3.5 g/dL. The patients were divided into two groups. In group 1 (n = 10), fasting serum Lp(a) and albumin were measured before, after repeated infusion of 20% albumin 100 mL three times per week for 2 weeks, and 4 weeks after withdrawal of albumin infusion. In group 2 (n = 10), serum albumin and Lp(a) were measured similarly without albumin infusion. C-reactive protein was monitored in both group as an indicator of acute-phase reactant. Serum Lp(a) was also measured in 20 age- and sex-matched normal controls. Apolipoprotein(a) [apo(a)] phenotype was determined in all the subjects. CAPD patients as a whole (n = 20; median, 70.2 mg/dL; interquartile range, 45.0 to 86.2 mg/dL) had higher serum Lp(a) than normal controls (n = 20; median, 9.9 mg/dL; interquartile range, 2.4 to 24.3 mg/dL) (P < 0.0001), although the distribution of apo(a) phenotype was similar. Serum albumin in group 1 increased from 2.6+/-0.5 g/dL to 3.5+/-0.6 g/dL (P < 0.0005) at the end of repeated infusion of albumin, whereas serum Lp(a) decreased from 73.7 mg/dL (range, 43.2 to 89.0 mg/dL) to 25.6 mg/dL (range, 10.7 to 71.7 mg/dL) (P < 0.01). Four weeks after withdrawal of albumin infusion, serum albumin decreased again to 2.9+/-0.5 g/dL (P < 0.001), whereas serum Lp(a) increased to 65.2 mg/dL (range, 43.3 to 106.0 mg/dL) (P < 0.05). Serum albumin in group 2 was 2.8+/-0.6 g/dL, 3.0+/-0.4 g/dL, and 2.9+/-0.7 g/dL, respectively. The change of serum Lp(a) was not significant (67.0 mg/dL [range, 46.8 to 84.8 mg/dL], 62.8 mg/dL [range, 45.1 to 81.0 mg/dL], and 63.0 mg/dL [range, 44.7 to 74.0 mg/dL]). C-reactive protein was stable during the study period in both groups. These findings support the hypothesis that hypoalbuminemia is one of the important trigger factors in the elevation of serum Lp(a) in CAPD patients. PMID- 9328366 TI - Patient assessments of adequacy of dialysis and protein nutrition. AB - Nephrologists closely monitor biochemical measurements of adequacy of dialysis and protein nutrition, but little is known about how patients assess adequacy. We sought (1) to determine how hemodialysis patients assess adequacy and (2) to compare patient assessments with objective measures of adequacy. We performed a cross-sectional interview study of 145 patients from two chronic hemodialysis units. Using a structured questionnaire, we asked subjects to assess the amount of dialysis and protein nutrition they were getting. Objective measures of amount of dialysis (Kt/V) and protein nutrition (albumin) were obtained from chart abstraction. We found that only 5% of all subjects thought they were getting less dialysis than they needed, yet 41% were actually receiving inadequate dialysis (Kt/V <1.2). Even among the 60 subjects with Kt/V less than 1.2, only 5% thought they were getting less dialysis than they needed. Similarly, 21% of all subjects thought they were getting less protein nutrition than they needed, yet 28% had inadequate protein nutrition levels (albumin <3.5 g/L). Even among the 41 subjects with albumin less than 3.5 g/L, only 20% thought they were getting less protein nutrition than they needed. In conclusion, patient assessments of adequacy differ greatly from the objective measures that nephrologists use to assess adequacy. Most patients with Kt/V less than 1.2 or albumin less than 3.5 g/L think they are getting adequate dialysis and protein nutrition. Understanding how patients assess adequacy may be an important step in developing interventions to improve the adequacy of dialysis and protein nutrition. PMID- 9328367 TI - Bacterial endocarditis in hemodialysis patients. AB - Infective endocarditis (IE) is one of the most serious complications of bacteremia. Hemodialysis patients with prosthetic vascular access devices such as dual-lumen cuffed venous catheters and polytetrafluoroethylene (PTFE) grafts are at increased risk for bacteremia compared with patients with primary arteriovenous fistulae (PAVF). We present 20 cases of IE in hemodialysis patients seen at our institution over a 7-year period. Eight patients had PTFE grafts, 11 had dual-lumen cuffed catheters, and one had a PAVF. All patients underwent transthoracic echocardiography (TTE) and 16 patients also received transesophageal echocardiography (TEE). The Duke Criteria were used to characterize patients as definite or possible IE. Organisms were Staphylococcus aureus (55%), Staphylococcus epidermidis (25%), and Enterococcus sp (10%). The mitral valve (50%) was the most commonly affected valve, followed by aortic (30%) and right-sided IE (25%). Valve replacement was performed in five patients, of whom three survived hospitalization. The overall mortality rate for the series was 30%. All the dual-lumen cuffed catheters were removed, while only two of the eight PTFE grafts were removed. Four of the six patients who died had PTFE grafts, of which one was judged to be infected and removed. Echocardiography was essential to the diagnosis of IE. Vegetations were found in 50% by TTE and 81% by TEE. Six patients with no vegetations on TTE were found to have vegetations on TEE. PMID- 9328368 TI - Care pathway reduces hospitalizations and cost for hemodialysis vascular access surgery. AB - Hemodialysis vascular access-related hospitalizations account for more than 20% of United States end-stage renal disease (ESRD) hospitalizations, with an annual cost approximating $675 million. Limiting access-related costs while delivering similar degrees of quality care thus would enhance alternative utilization of ESRD funding. We implemented a vascular access care pathway emphasizing coordinated patient evaluation and outpatient surgery to determine whether such an intervention affected outcomes associated with vascular access surgery. Data examining hospitalization and vascular access surgery charges, complications, and patient satisfaction (determined by questionnaire) were analyzed, comparing patients who underwent vascular access surgery in 1994 and 1995 as inpatients (non-care pathway patients) and patients who underwent vascular access surgery via the care pathway in 1995. Inpatient days declined in 1995 (1994: 582 days; 1995: 85 days; P < 0.03) and the average charges per patient for the care pathway cohort were significantly less than charges per patient in 1994 and charges for non-care pathway patients in 1995 (1994 patients: $10,524 +/- $5,209; 1995 non care pathway patients: $11,196 +/- $5,806; 1995 care pathway patients: $4,686 +/- $2,912/patient; P < 0.02). Incidence rates for major (life-threatening) complications were not significantly different between 1994 patients and care pathway patients in 1995. However, the 1995 non-care pathway patients had a higher incidence of major complications (15.4%). Forty-seven repeat access procedures were performed in 29 patients in 1994 versus 35 repeat access procedures in 22 care pathway patients in 1995, and 12 repeat access procedures were performed in eight non-care pathway patients in 1995. Finally, a majority of the patients entered into the care pathway who responded to a survey stated that they were satisfied with access surgery via the care pathway. These data suggest that a vascular access care pathway can reduce hospital days and costs while achieving acceptable outcomes for access surgery. PMID- 9328369 TI - Markers of masked iron deficiency and effectiveness of EPO therapy in chronic renal failure. AB - Recombinant erythropoietin (rHuEPO) is well established in the management of anemia of chronic renal disease. However, a number of clinical issues, including the best laboratory indicators of an imminent marrow response to rHuEPO replacement, the ideal measurements to detect masked iron deficiency, and optimal methods of iron replacement, remain unanswered. To investigate these issues, studies were performed in anemic chronic hemodialysis patients. A number of standard hematologic measurements in addition to automated reticulocyte counts (Sysmex R-1000) and serum transferrin receptors (TfR) were obtained in these patients. A response to initiation of rHuEPO administration could be predicted if the serum TfR concentration was less than 6 mg/L (normal, 3.8 to 8.5 mg/L). In patients on rHuEPO, an imminent hemoglobin response to an increased rHuEPO dose could be predicted after 1 week based on a greater than 20% increase from baseline in the serum TfR or absolute reticulocyte count, with a sensitivity of 92%. In patients on rHuEPO replacement with serum ferritin levels greater than 30 microg/L, none of the panel of tests, including serum TfR, reliably detected masked iron deficiency. In a long-term study over 5 months in patients on a stable maintenance dose of EPO, a gradual decline in total body iron occurred, even in subjects with initial adequate iron stores, and despite taking 50 mg elemental iron daily as oral ferrous sulphate. The serum TfR is useful for predicting a hemoglobin response when initiating rHuEPO therapy, and combined with automated reticulocyte counting it is valuable for predicting a hemoglobin response when increasing the dose of rHuEPO. The serum TfR loses its specificity for detecting tissue iron deficiency in patients on maintenance rHuEPO therapy because of increased erythropoiesis, which itself raises serum TfR levels. PMID- 9328370 TI - Impact of capitation on free-standing dialysis facilities: can you survive? AB - Proposed changes in the Medicare reimbursement method for end-stage renal disease (ESRD) patients prompted us to study the total cost of caring for the ESRD patients in northeast Indiana over a 1-year period. We hoped to ascertain the actual cost of caring for patients treated with different modalities, determine if we could compete in a capitated environment, and identify areas in which we might reduce these expenses. Six patients new to dialysis and 29 patients already receiving treatment underwent follow-up evaluation for 1 year. We tracked their cost of care for 1 year in the outpatient setting as well as in the hospital. We found the cost of caring for all patients was $43,044 per year. Patients new to dialysis cost $3,164 more to care for than patients already receiving dialysis treatment. Hospitalization expense was the primary component of that difference. Continuous ambulatory peritoneal dialysis (CAPD) patients were $14,570 less costly per year to care for than hemodialysis patients. This differential primarily related to decreased hospitalization. Vascular access expenses were a major component of both the outpatient and inpatient cost for hemodialysis patients. Our yearly expenditures for all patients compared with suggested capitated Medicare reimbursement rates suggested that our program could be successful in a new reimbursement model. Several areas of possible cost reduction were identified. PMID- 9328371 TI - Living kidney donation: a survey of professional attitudes and practices. AB - Living donation is an option for meeting the needs of patients with end-stage renal disease. We surveyed kidney transplant professionals to understand their attitudes, opinions, and practices regarding living donation and to generate a rough estimate of the national potential for living related kidney donation. Although this sample of health practitioners expressed strong support for living donation, their actual professional practice does not necessarily reflect such support. Given the disparity between support and practice, we recommend that leaders in the kidney transplant community focus on several concrete objectives to optimize living donation: estimate the underlying potential for living donation; identify and implement best demonstrated practices for making the living donation request; explore and respond to the ethical issues that surround living donation; and address the needs of both donors and recipients. PMID- 9328372 TI - Acute renal failure due to indinavir crystalluria and nephrolithiasis: report of two cases. AB - Two patients with oliguric acute renal failure (ARF) attributed to crystalluria and nephrolithiasis with obstructive uropathy caused by the human immunodeficiency virus protease inhibitor indinavir are described. In both patients, ARF resolved with administration of intravenous fluids. One patient required urologic intervention to relieve bilateral ureteral obstruction. PMID- 9328373 TI - Marked dilutional acidosis complicating management of right ventricular myocardial infarction. AB - Dilutional acidosis is a poorly recognized cause of metabolic acidosis. Indeed, the prevailing view has been that even massive expansion of the extracellular fluid volume with non-bicarbonate-containing solutions would not lead to clinically significant hypobicarbonatemia. We describe the development of marked dilutional acidosis as a complication of management of right ventricular myocardial infarction. The pathogenesis, clinical significance, prevention, and treatment of the entity are discussed. PMID- 9328374 TI - Treatment of acute methanol intoxication with hemodialysis using an ethanol enriched, bicarbonate-based dialysate. AB - A patient poisoned with methanol was successfully hemodialyzed with an ethanol enriched, bicarbonate-based dialysate. Along with a concomitant intravenous infusion of ethanol, the ethanol-enriched dialytic procedure was able to maintain an intradialytic plasma ethanol level of 80 to 102 mg/dL. The patient recovered without any sequelae of methanol intoxication. PMID- 9328375 TI - Spurious hypophosphatemia in a patient with multiple myeloma. AB - We report a patient with multiple myeloma and a prolonged history of hypophosphatemia who had remained asymptomatic. Extensive evaluation for a cause, including the search for a renal tubular disorder, oncogenous osteomalacia, or a parathyroid hormone (PTH)-related protein was unproductive. Renal biopsy showed no evidence of myeloma kidney. Subsequent mixing of the immunoglobulin G (IgG) fraction from the patient's serum with normal human serum, confirmed that the observed hypophosphatemia was spurious, resulting from interference of large amounts of an abnormal IgG with a standard automated laboratory assay for phosphate. PMID- 9328376 TI - Hypertension in hemodialysis patients: more questions than answers. PMID- 9328377 TI - Renal ultrasonography: a procedure for nephrologists. PMID- 9328378 TI - Granulomatous interstitial nephritis: drug hypersensitivity, infection, or sarcoidosis? PMID- 9328379 TI - Is there a hepatitis C virus-associated membranoproliferative glomerulonephritis? PMID- 9328380 TI - Risk adjustment of the postoperative mortality rate for the comparative assessment of the quality of surgical care: results of the National Veterans Affairs Surgical Risk Study. AB - BACKGROUND: The National Veterans Affairs Surgical Risk Study was designed to collect reliable, valid data on patient risk and outcomes for major surgery in the Veterans Health Administration and to report comparative risk-adjusted postoperative mortality rates for surgical services in Veterans Health Administration. STUDY DESIGN: This cohort study was conducted in 44 Veterans Affairs Medical Centers. Included were 87,078 major noncardiac operations performed under general, spinal, or epidural anesthesia between October 1, 1991, and December 31, 1993. The main outcomes measure was all-cause mortality within 30 days after the index procedure. Multivariable logistic regression risk adjustment models for all operations and for eight surgical subspecialties were developed. Risk-adjusted surgical mortality rates were expressed as observed-to expected ratios and were compared with unadjusted 30-day postoperative mortality rates. RESULTS: Patient risk factors predictive of postoperative mortality included serum albumin level, American Society of Anesthesia class, emergency operation, and 31 additional preoperative variables. Considerable variability in unadjusted mortality rates for all operations was observed across the 44 hospitals (1.2-5.4%). After risk adjustment, observed-to-expected ratios ranged from 0.49 to 1.53. Rank order correlation of the hospitals by unadjusted and risk adjusted mortality rates for all operations was 0.64. Ninety-three percent of the hospitals changed rank after risk adjustment, 50% by more than 5 and 25% by more than 10. CONCLUSIONS: The Department of Veterans Affairs has successfully implemented a system for the prospective collection and comparative reporting of risk-adjusted postoperative mortality rates after major noncardiac operations. Risk adjustment had an appreciable impact on the rank ordering of the hospitals and provided a means for monitoring and potentially improving the quality of surgical care. PMID- 9328381 TI - Risk adjustment of the postoperative morbidity rate for the comparative assessment of the quality of surgical care: results of the National Veterans Affairs Surgical Risk Study. AB - BACKGROUND: The National Veterans Affairs Surgical Risk Study was designed to collect reliable, valid data on patient risk and outcomes for major surgery in the Veterans Health Administration and to report comparative risk-adjusted postoperative mortality and morbidity rates for surgical services in the Veterans Health Administration. STUDY DESIGN: This was a cohort study conducted at 44 Veterans Affairs Medical Centers closely affiliated with university medical centers. Included were 87,078 major noncardiac operations performed under general, spinal, or epidural anesthesia between October 1, 1991, and December 31, 1993. The main outcomes measures in this report are 21 postoperative adverse events (morbidities) occurring within 30 days after the index procedure. Multivariable logistic regression risk-adjustment models for all operations and for eight surgical subspecialties were developed. RESULTS: Patient risk factors predictive of postoperative morbidity included serum albumin level, American Society of Anesthesia class, the complexity of the operation, and 17 other preoperative risk variables. Wide variation in the unadjusted rates of one or more postoperative morbidities for all operations was observed across the 44 hospitals (7.4-28.4%). Risk-adjusted observed-to-expected ratios ranged from 0.49 to 1.46. The Spearman rank order correlation between the ranking of the hospitals based on unadjusted morbidity rates and risk-adjusted observed-to-expected ratios for all operations was 0.87. There was little or no correlation between the rank order of the hospitals by risk-adjusted morbidity and risk-adjusted mortality. CONCLUSIONS: The Department of Veterans Affairs has successfully implemented a system for the prospective collection and comparative reporting of postoperative mortality and morbidity rates after major noncardiac operations. Risk adjustment had only a modest effect on the rank order of the hospitals. PMID- 9328382 TI - Validating risk-adjusted surgical outcomes: site visit assessment of process and structure. National VA Surgical Risk Study. AB - BACKGROUND: Risk-adjusted mortality and morbidity rates are often used as measures of the quality of surgical care. This study was conducted to determine the validity of risk-adjusted surgical morbidity and mortality rates as measures of quality of care by assessing the process and structure of care in surgical services with higher-than-expected and lower-than-expected risk-adjusted 30-day mortality and morbidity rates. STUDY DESIGN: A structural survey of 44 Veterans Affairs Medical Center surgical services and site visits to 20 surgical services with higher-than-expected and lower-than-expected risk-adjusted outcomes were conducted. Main outcome measures included assessment of technology and equipment, technical competence of staff, leadership, relationship with other services, monitoring of quality of care, coordination of work, relationship with affiliated institutions, and overall quality of care. RESULTS: Surgical services with lower than-expected risk-adjusted surgical morbidity and mortality rates had significantly more equipment available in surgical intensive care units than did services with higher-than-expected outcomes (4.3 versus 2.9, p < 0.05). Site visitor ratings of overall quality of care were significantly higher for surgical services with lower-than-expected morbidity and mortality rates (6.1 versus 4.5 for high outliers, p < 0.05); technology and equipment were rated significantly better among low-outlier services (7.1 versus 4.8 for high outliers, p < 0.001). Masked site-visit teams correctly predicted the outlier status (high versus low) of 17 of the 20 surgical services visited (p < 0.001). CONCLUSIONS: Significant differences in several dimensions of process and structure of the delivery of surgical care are associated with differences in risk-adjusted surgical morbidity and mortality rates among 44 Veterans Affairs Medical Centers. PMID- 9328383 TI - Longterm followup (12-15 years) of a randomized controlled trial comparing Bassini-Stetten, Shouldice, and high ligation with narrowing of the internal ring for primary inguinal hernia repair. AB - BACKGROUND: Shouldice repair for primary inguinal hernia is reported to have better results than classic Bassini-type repairs. The indirect inguinal hernia with a firm posterior wall is often assumed to be adequately treated by high ligation and ring narrowing. STUDY DESIGN: This double randomized controlled trial compared high ligation and ring narrowing with Bassini-Stetten repair for the indirect inguinal hernia with a firm posterior wall, and Shouldice with Bassini-Stetten repair for the inguinal hernia with a weakened posterior wall, direct or indirect. This report focuses on longterm (12-15 years) recurrence rates. RESULTS: From July 1980 to May 1983, 102 indirect primary inguinal hernias with a firm posterior wall (group I) and 263 primary inguinal hernias with a weakened posterior wall (group II) were included. By 1995, 89 patients with 100 hernia repairs had died, and for 30 repairs the patients could not be located. In 41 hernia repairs, a recurrence had been established previously. Of the remaining 194 hernia repairs, followup was updated by physical examination in 179 (92%) and by telephone interview in 15 (8%). A total of 83 recurrences were recorded, 42% of which were asymptomatic at the time of diagnosis. Seventy-three percent of the recurrences happened > 2 years after the operation. The life-table method showed the following longterm (12-15 years) recurrence rates: group I, Bassini-Stetten 33% versus ring narrowing 34%; group II, Bassini-Stetten 32% versus Shouldice 15% (p = 0.033). CONCLUSIONS: The Shouldice is the best type of hernia repair, although the 15% recurrence rate is high. Bassini-Stetten and high ligation with ring narrowing are inadequate repairs, regardless of the type of hernia. PMID- 9328384 TI - Attenuation of ischemic liver injury by monoclonal anti-endothelin antibody, AwETN40. AB - BACKGROUND: Enhanced production of endothelin-1 (ET-1), vasoconstrictive 21 amino acids produced by endothelial cells during ischemia and after reperfusion of the liver, is known to cause sinusoidal constriction and microcirculatory disturbances, which lead to severe tissue damage. Using a 2-hour hepatic vascular exclusion model in dogs, we tested our hypothesis that neutralization of ET-1 by monoclonal anti-ET-1 and anti-ET-2 antibody (AwETN40) abates vascular dysfunction and ameliorates ischemia/reperfusion injury of the liver. STUDY DESIGN: After skeletonization, the liver was made totally ischemic by cross-clamping the portal vein, the hepatic artery, and the vena cava (above and below the liver). Veno venous bypass was used to decompress splanchnic and inferior systemic congestion. AwETN40, 5 mg/kg, was administered intravenously 10 minutes before ischemia (treatment group, n = 5). Nontreated animals were used as controls (control group, n = 10). Animal survival, hepatic tissue blood flow, liver function tests, total bile acid, high-energy phosphate, ET-1 levels, and liver histopathology were studied. RESULTS: Treatment with AwETN40 improved 2-week animal survival from 30% to 100%. Hepatic tissue blood flow after reperfusion was significantly higher in the treatment group. The treatment significantly attenuated liver enzyme release, total bile acid, and changes in adenine nucleotides. Immunoreactive ET-1 levels in the hepatic venous blood of the control group showed a significant increase and remained high for up to 24 hours after reperfusion. Histopathologic alterations were significantly lessened in the treatment group. CONCLUSIONS: These results indicate that ET-1 is involved in ischemia/reperfusion injury of the liver, which can be ameliorated by the monoclonal anti-ET-1 and anti-ET-2 antibody AwETN40. PMID- 9328386 TI - Delayed gastric emptying after standard pancreaticoduodenectomy versus pylorus preserving pancreaticoduodenectomy: an analysis of 200 consecutive patients. AB - BACKGROUND: It has been suggested that pylorus-preserving pancreaticoduodenectomy (PPPD) is associated with a high incidence of delayed gastric emptying and consequently with a prolonged hospital stay compared with standard Whipple's resection. The aim of this prospective study was to evaluate whether the incidence of delayed postoperative gastric emptying was different after both procedures. STUDY DESIGN: From 1989 to 1996, 200 consecutive patients underwent pancreatic head resection (100 standard pancreaticoduodenectomy [PD]; 100 PPPD). The groups were compared with regard to patient characteristics, operative indices, postoperative morbidity, hospital stay, and mortality. Delayed gastric emptying was defined as nasogastric suction for > or = 10 days or delay of regular diet until > 14 days postoperatively. RESULTS: Operative time and blood loss were higher for PD: 6 versus 4.8 hours (p < 0.0001) and 1,580 versus 1,247 mL (p < 0.001), respectively. Postoperative morbidity was 48% after PD and 44% after PPPD (not significant [NS]). Hospital mortality was 6% and 1% after PD and PPPD, respectively (NS). Delayed gastric emptying occurred in 34 patients after PD and in 37 after PPPD (NS). Median days of gastric suction was 3 versus 6 days for PD and PPPD (p < 0.0001). A regular diet was tolerated after a median of 10 and 11 days for PD and PPPD, respectively (NS). Postoperative hospital stay was shorter for patients who underwent PPPD: 20 versus 18 days (p < 0.03). Patients with intraabdominal complications (n = 52) showed a higher incidence of delayed gastric emptying (p < 0.0001). CONCLUSIONS: Our results show that PPPD is a safe procedure associated with less operative time and blood loss than PD. After PPPD, patients require longer postoperative nasogastric intubation than after PD, suggesting some form of early postoperative gastric stasis. There is, however, no difference in the incidence of delayed gastric emptying or the first postoperative day on which a regular diet is tolerated between these surgical procedures. Intraabdominal complications are major risk factors for delayed gastric emptying. PMID- 9328385 TI - Small-molecule selectin inhibitor protects against liver inflammatory response after ischemia and reperfusion. AB - BACKGROUND: The selectin family of adhesion molecules plays a key role in the neutrophil-mediated injury observed after ischemia and reperfusion. In our study, we investigated the effects of TBC-1269, a novel small-molecule, nonoligosaccharide inhibitor of P-, E-, and L-selectin binding, in the liver inflammatory response after 90 minutes of warm ischemia. STUDY DESIGN: Total liver ischemia was produced in Sprague-Dawley rats for 90 minutes using an extracorporeal portosystemic shunt. The animals were divided into five groups including: the sham (group 1), ischemic control (group 2) receiving only the vehicle, and the treated groups receiving TBC-1269 at a dose of 25 mg/kg at different times of administration: 15 minutes before reperfusion (group 3), at reperfusion (group 4), and 15 minutes after reperfusion (group 5). The following indices were analyzed: 7-day survival, liver injury tests, liver tissue myeloperoxidase as an index of neutrophil infiltration, and liver histology. RESULTS: TBC-1269 treated groups experienced a significant increase in survival compared with controls. Best overall survival, 70%, was observed when TBC-1269 (Texas Biotechnology Corporation, Houston, TX) was administered 15 minutes before reperfusion (p < 0.05). This group also showed a marked decrease (p < 0.05) in liver enzyme levels at 6 hours after reperfusion. Neutrophil migration was also significantly ameliorated (81%), as reflected by decreased myeloperoxidase levels. We observed improved histologic damage scores in the treated group compared with controls (p < 0.05). CONCLUSIONS: A small-molecule selectin inhibitor (TBC-1269) had a protective effect in livers subjected to 90 minutes of warm hepatic ischemia and 6 hours of reperfusion by decreasing neutrophil infiltration, migration and subsequent tissue damage. The best protective effect was achieved when the compound was administered 15 minutes before reperfusion. These findings offer a new therapeutic alternative for protection against ischemia and reperfusion injury. PMID- 9328387 TI - Role of esophageal body function in gastroesophageal reflux disease: implications for surgical management. AB - BACKGROUND: Effective esophageal peristalsis is a major determinant of esophageal clearance function. The relation of esophageal body function with a mechanically defective lower esophageal sphincter and the development of esophageal mucosal injury in patients with gastroesophageal reflux disease is unclear. STUDY DESIGN: We analyzed the relations among the manometrically determined esophageal clearance function, lower esophageal sphincter dysfunction, esophageal acid exposure, and the presence and severity of esophageal mucosal injury in patients with gastroesophageal reflux disease. Normal values for the manometric assessment of esophageal clearance function were established in 50 normal volunteers and subsequently applied to 160 symptomatic patients with increased esophageal exposure to gastric juice and various grades of esophageal mucosal injury (no minimal surgery, esophagitis, stricture, and Barrett's esophagus). RESULTS: Defective clearance function was present in 47.5% of the patients; a defective lower esophageal sphincter was documented in 63.1%. Compromised esophageal clearance function was significantly more common in patients with a defective lower esophageal sphincter than in those with normal sphincter function (55% versus 33.8%). Esophageal acid exposure time and the prevalence and severity of esophageal mucosal injury were highest in patients with a defective sphincter and compromised clearance function. CONCLUSIONS: These data show that esophageal motor function deteriorates with increasing severity of mucosal injury. This appears to be due to persistent reflux of gastric juice across a mechanically defective lower esophageal sphincter. This may influence the choice and outcome of antireflux surgery. Surgical correction of a mechanically defective sphincter before the loss of esophageal body function is advocated. PMID- 9328389 TI - Clinicopathologic significance of the expression of mutated p53 protein and the proliferative activity of cancer cells in patients with esophageal squamous cell carcinoma. AB - BACKGROUND: The aim of this study was to investigate the relation between the expression of mutated p53 protein and the proliferative activity of cancer cells in esophageal squamous cell carcinoma. In addition, the clinical and biologic significance of p53 status and the proliferative activity of cancer cells were evaluated in these patients. STUDY DESIGN: Samples of esophageal tumors from 94 patients were subjected to immunohistochemical staining with a monoclonal antibody against p53 and with the monoclonal antibody Ki-67. The immunoreactivity against p53 and the proliferative activity of cancer cells were compared with the clinicopathologic findings in each sample. Prognostic factors including p53 status and Ki-67 labeling index (LI; percentage of Ki-67-immunostained cells) were evaluated for 81 surviving patients by univariate and multivariate analysis. RESULTS: The mean Ki-67 LI of 50 p53-positive patients was higher than that of 44 p53-negative patients (p = 0.009). The Ki-67 LI increased according to the progression of tumors. Overexpression of mutated p53 protein was observed in 40.9% of tumors that invaded to the submucosa, and this percentage was not significantly changed in tumors with invasion to the adventitia. Metastases to the regional lymph nodes were observed in 3 of 22 patients with tumors that invaded to the submucosa, and these 3 tumors had both over-expression of mutated p53 protein and high Ki-67 LI. In 81 surviving patients, only lymph node metastasis (p = 0.045) and the curability of tumors (p < 0.001) were identified as independent prognostic factors by multivariate analysis. CONCLUSIONS: Overexpression of mutated p53 protein is detected in the early stage of esophageal cancer. This mutated p53 protein may not play an important role for tumor invasion. When tumor invasion is limited to the submucosa, cancer cells that overexpress mutated p53 protein may acquire high proliferative activity. Such cells might have considerable potential for metastasis to the lymph nodes. PMID- 9328388 TI - Management of thyroid cancer of follicular cell origin: Gundersen/Lutheran Medical Center, 1969-1995. AB - BACKGROUND: Most reports regarding the treatment of thyroid cancer originate from university referral centers. In this article, we report our experience in managing thyroid cancer of follicular cell origin at a non-university institution over a 26-year period. STUDY DESIGN: We reviewed the medical records of all patients treated for thyroid cancer at the Gundersen/Lutheran Medical Center from 1969 to 1995. Histologic types, demographic and clinical characteristics, laboratory results, treatment, complications, and followup observations were tabulated. Risk was assigned according to the age, presence of distant metastasis, extent of the primary tumor, and site of the primary tumor (AMES) staging system. RESULTS: The histologic classification was as follows: papillary, 139; follicular, 24; Hurthle cell, 14; and anaplastic, 11. Low-risk lesions were identified in 96%, 79%, and 71% of the patients with papillary, follicular, and Hurthle cell (collectively designated differentiated) carcinoma, respectively. We treated 60% of our patients with differentiated thyroid cancer with near-total or total thyroidectomy. Clinically involved cervical lymph nodes were removed singly or by modified neck dissection. We frequently ablated thyroid remnants after operation with 29.9 mCi (1,110 MBq) of 131I, after which we treated the patient with suppressive doses of levothyroxine. Patients were evaluated yearly with thyroglobulin measurements and, in some high-risk patients, with total-body 131I scans. Cancer recurred in 13%, 8%, and 7% of our patients with papillary, follicular, and Hurthle cell carcinoma, respectively. Only three patients died of differentiated thyroid cancer; eight are alive with malignancy. In anaplastic thyroid cancer, cervical lymph node metastases, local invasion, and distant metastases were present in 18%, 64%, and 45% of patients at the time of initial evaluation. Total or near-total thyroidectomy was possible in only four of nine patients treated surgically. External radiation (11 patients) and chemotherapy (two patients) were used. Additional metastases developed in 45% of the patients, and nine patients died within a year. Permanent hypoparathyroidism or hoarseness complicated 2.7% of the thyroid operations. CONCLUSIONS: Although our followup was relatively short, the results of treating thyroid cancer by general surgeons at a nonuniversity hospital compare favorably with results obtained from university referral centers. PMID- 9328390 TI - Does vaginal cuff closure decrease the infectious morbidity associated with abdominal hysterectomy? AB - BACKGROUND: Infectious morbidity after total abdominal hysterectomy includes fever (31%) and antibiotic administration (45%). Whether vaginal cuff closure reduces postoperative infectious morbidity remains unresolved. STUDY DESIGN: We reviewed the records of 172 consecutive abdominal hysterectomies for nonmalignant disease performed at an inner-city hospital. We identified potential risk factors for infectious morbidity by univariate analysis and determined adjusted odds ratios by multiple logistic regression analysis. RESULTS: The open vaginal cuff technique was associated with an increased risk of wound infection. Use of prophylactic antibiotics was associated with a decreased risk of febrile morbidity and a decreased risk of prolonged hospitalization. Body weight in the heaviest quartile was associated with increased risk of wound infection, increased risk of prolonged hospitalization, and decreased risk of postoperative vaginal cuff granulation tissue. Older age was associated with an increased risk of prolonged hospitalization. CONCLUSIONS: Closure of the vaginal cuff and use of prophylactic antibiotics at total abdominal hysterectomy were associated with decreased infectious morbidity in a high-risk population. PMID- 9328391 TI - The future is now. PMID- 9328392 TI - Decompression of the gastrointestinal tract after esophageal operations: how to do it. PMID- 9328393 TI - Leaning spleen: a new approach to laparoscopic splenectomy. PMID- 9328394 TI - Surgical education opportunities in for-profit hospitals. PMID- 9328395 TI - Laparoscopic herniorrhaphy revisited. PMID- 9328396 TI - Laparoscopic herniorrhaphy revisited. PMID- 9328397 TI - Laparoscopic herniorrhaphy revisited. PMID- 9328398 TI - Laparoscopic herniorrhaphy revisited. PMID- 9328431 TI - Delays in malignant tumor development in transgenic mice by forced epidermal keratin 10 expression in mouse skin carcinomas. AB - The keratin cytoskeleton is formed in different epidermal compartments by distinct polypeptides. Basal, proliferative keratinocytes express keratin (K) 5 and K14, whereas, suprabasal, post-mitotic keratinocytes express K1 and K10. Changes in this keratin pattern have been found to occur in hyperproliferative skin disorders and, in particular, throughout mouse epidermal carcinogenesis. Whereas some keratins not found in normal epidermis (K6, K16, K13, and K8) are induced at different stages of tumor development, K1 and K10 expression is lost. To determine whether K1 and K10 loss is just a consequence of the altered differentiation program or an event required for tumor progression, we generated transgenic mice carrying the human keratin 10 gene (hK10) under the control of a bovine keratin 6 gene regulatory region, which is silent in normal skin but is induced and drives transgene expression in hyperproliferative skin keratinocytes and, therefore, in skin tumors. Transgenic animals subjected to a complete carcinogenesis protocol developed tumors that contained various amounts of transgenic hK10. Although no significant difference was found in tumor number or malignancy, tumor onset was significantly delayed in transgenic mice, indicating that the presence of K10 actually impairs tumor development. PMID- 9328432 TI - Induction of PA2.26, a cell-surface antigen expressed by active fibroblasts, in mouse epidermal keratinocytes during carcinogenesis. AB - The monoclonal antibody PA2.26, produced against mouse epidermal keratinocytes transformed with 7,12-dimethylbenz[a]anthracene (DMBA), recognizes a 43- to 47 kDa cell-surface protein that was absent from non-tumorigenic epidermal keratinocytes but present in transformed epidermal cell lines as well as cultured normal fibroblasts. In vivo, the antigen was absent from normal epidermis but induced in basal-like epidermal keratinocytes and dermal fibroblasts during tissue regeneration after wounding and treatment with the tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA). The PA2.26 protein was also expressed in basal-like cells of differentiated papillomas and carcinomas generated in mice treated with DMBA and TPA. In addition, the antigen was abundantly synthesized by stromal cells of the tumors. These results suggest that PA2.26 antigen is involved in reactive processes during skin remodeling and carcinogenesis. PMID- 9328433 TI - Comparison of chemical carcinogen skin tumor induction efficacy in inbred, mutant, and hybrid strains of mice: morphologic variations of induced tumors and absence of a papillomavirus cocarcinogen. AB - Chemical carcinogen induction of skin tumors in mice was investigated to determine (i) if tumor induction efficacy was modified by single gene mutations, (ii) if the histologic types of the tumors varied with these mutations, and (iii) if a novel papillomavirus was involved as a cocarcinogen. A two-stage carcinogenesis protocol (7,12-dimethylbenz[a]anthracene followed by 12-O tetradecanoylphorbol-13-acetate) was used to induce papillomas in 14 inbred, two hybrid, and 15 other genetic stocks of mice with inherited, single-gene mutations causing skin abnormalities. Histopathological, immunohistochemical, and Southern blot analyses were performed to determine tumor type and to detect the presence of papillomaviruses. The histologic types of tumors induced included early follicular papillomas, mixed papillomas, exophytic papillomas, hyperplastic papillomas, fibropapillomas, squamous cell carcinomas, and mast cell tumors. The efficacy of tumor induction was influenced by strain background, as seen by the clustering of mice into high-, intermediate-, and nonresponding groups. Similarly, tumor induction efficacy was affected by specific mutant genes that cause skin abnormalities. No evidence of papillomavirus structural antigens or viral genomic DNA was identified in 547 induced tumors. These observations indicate that numerous modifier genes but not papillomaviruses are involved in cutaneous chemical carcinogenesis. PMID- 9328434 TI - Analysis of the E-cadherin and P-cadherin promoters in murine keratinocyte cell lines from different stages of mouse skin carcinogenesis. AB - We previously isolated the 5' upstream sequences of the mouse P-cadherin gene, in which putative binding sites for several transcription factors were identified between nt-101 and +30. In the study reported here, the promoter activity of the postulated 5' cis-acting sequences of the P-cadherin promoter, and the activity of the proximal E-cadherin promoter were investigated in several murine keratinocyte cell lines showing different levels of P- and E-cadherin expression as well as different morphology and tumorigenic behavior. Cell-type specificity and optimal activity of P-cadherin expression in murine keratinocytes was conferred by 5' sequences located between nt -200 and +30, and the GC-rich region (nt -101 to +80) and a CCAAT box element (nt -65) had a major regulatory role. The cell-type specificity of the E-cadherin promoter, on the other hand, was mediated by a combination of positive regulatory elements, a GC-rich region (nt 58 to -24), and a CCAAT box (nt -65) and repressor elements inside the E-pal sequence. Interestingly, the maximum repressor effect of the E-pal element was observed in non-expressing undifferentiated spindle cells. In vitro binding studies indicated that the GC-rich region of the P-cadherin promoter was mainly recognized by Sp1-related nuclear factors, whereas both AP2- and Sp1-related factors were involved in the interaction of the GC-rich region of the E-cadherin promoter. Common factors (probably related to the CP1 family) seemed also to be involved in the recognition of the CCAAT-box element of both the E- and P cadherin promoters, but additional specific factors participated in the interaction with the CCAAT box of the E-cadherin promoter. Our studies also support the hypothesis that loss or modification of some of the regulatory factors occurs during mouse skin tumor progression. PMID- 9328435 TI - Transplantation analyses of the immunogenicity of epidermal tumors generated in murine skin two-stage carcinogenesis protocols. AB - SSIN mice are very sensitive to tumor promoters in two-stage skin carcinogenesis protocols. It was recently reported that SSIN mice have fewer CD8+ T-cells than other strains of mice and develop a weaker cytotoxic T-cell response upon challenge with an allogeneic tumor transplant. The significance of this muted immune response to processes involved in two-stage carcinogenesis depends on the immunogenicity of the tumors generated in such protocols. Although they have low CD8+ T-cell contents, SSIN rejected a variety of subcutaneously transplanted allogeneic murine tumors. Analyses of the growth of primary papillomas derived from 7,12-dimethylbenz[a]anthracene-initiated/12-O-tetradecanoylphorbol-13-ac etate-promoted SSIN mice and then subcutaneously transplanted into triple deficient (bg-nu-xid), athymic nude and immune-competent and immunosuppressed SSIN mice revealed that few tumors took and tumor takes were not markedly influenced by the immumological status of the transplant recipient. Two tumor cell lines (RS1 and RS2) were derived from the transplantation studies and could be passaged in normal SSIN mice (H-2q haplotype). Both tumors were squamous cell carcinomas (SCCs) by the second in vivo passage and were rejected in allogeneic mice (BALB/c) but grew in FVB/N mice, a strain having the H-2q haplotype. Transplantation studies revealed that prior exposure to RS1 and RS2 did not prime SSIN mice to reject a subsequent tumor challenge. Three primary SCC tumors derived from SSIN mice in a two-stage carcinogenesis protocol also grew when subcutaneously transplanted in SSIN mice and could be serially passaged. Consequently, the epidermal SCCs that develop in two-stage carcinogenesis protocols appear to be nonimmunogenic. PMID- 9328436 TI - Retinoic acid signaling cascade in differentiating murine epidermal keratinocytes: alterations in papilloma- and carcinoma-derived cell lines. AB - The retinoic acid (RA) signaling pathway was investigated by transient transfection of a chloramphenicol acetyltransferase (CAT) reporter gene construct containing the RA response element (RARE) of the murine (m) RARbeta2 gene into murine primary epidermal keratinocytes (PEK), papilloma-derived SP1 cells, and carcinoma-derived 3P2 cells. Murine PEK transfected in a low-Ca2+ medium (0.05 mM Ca2+) exhibited a strong transactivation of the CATgene after exposure of the cells to 0.1 microM RA. Transactivation of the CATgene could, however, also be achieved by shifting RAREbeta2-transfected low-Ca2+ PEK to high-Ca2+ conditions (0.15-1.2 mM Ca2+). Concomitantly, the Ca2+ raise also led to the induction of both cellular retinol (ROL)-binding protein I (CRBPI) and cellular RA-binding protein II (CRABPII), whereas expression of cellular RA-binding protein I (CRABPI) was not observed. Moreover, induction of in vitro differentiation also activated the ROL-->RA converting enzyme system in PEK. These findings suggest the following sequence of events involved in the high Ca2+-mediated activation of RAREbeta2. First, high Ca2+ induces the synthesis of mCRBPI, which binds ROL released from retinyl ester stores and makes it accessible to the ROL-RA converting enzyme system. Enzymatically generated RA is taken over by mCRABPII and transported to the nucleus, where it acts as ligand for nuclear receptors, which complex with RAREbeta2 to activate the reporter gene. This hypothetical cascade of RA signaling was supported by our findings that inhibition of the ROL- >RA converting enzyme system by citral abolished the Ca2+-mediated transactivation of the CAT gene in a nontoxic manner. Studies in transformed murine cell lines revealed that Ca2+-induced activation of RAREbeta2 was essentially maintained in papilloma-derived SP1 cells, although all parameters of the Ca2+-dependent RAREbeta2 activation cascade were induced to a much lower extent. In contrast, strong RAREbeta2 activity was already observed in low-Ca2+ carcinoma-derived 3P2 cells. Low-Ca2+ 3P2 cells also expressed high levels of both mCRBPI and mCRABPII and possessed a highly active ROL-->RA converting enzyme system. Again, inhibition of the enzyme by citral abolished RAREbeta2 activity in low-Ca2+ 3P2 cells. Our data show that Ca2+-induced differentiation in cultured murine PEK entails a series of events that ultimately lead to the activation of RARE-containing genes. These properties are maintained in transformed epidermal keratinocytes. However, with increasing malignant potential of the cells, the respective signaling pathway becomes independent from a differentiation stimulus and leads to constitutive activation of RARE-controlled genes. PMID- 9328437 TI - Decreases in phorbol ester-induced papilloma development in v-Ha-ras transgenic TG.AC mice during reduced gene dosage of bcl-2. AB - We have demonstrated that induction of transgene expression in the v-Ha-ras transgenic TG.AC mouse is a critical event in skin tumorigenesis and that cutaneous papillomas arise from follicular epidermis after treatment with chemical carcinogens. The sensitivity of TG.AC mice to skin tumorigenesis, coupled with their low incidence of spontaneous skin tumors, makes this strain a good model for identifying carcinogens and for investigating the roles that other genes may play in the development of skin neoplasia. To investigate the possible involvement of the bcl-2 gene in skin tumorigenesis in the TG.AC mouse, we crossed heterozygous bcl-2-knockout mice (C57BI/6, 129 background) with TG.AC mice (FVB/N background). Female mice were genotyped by using a neo cassette to identify bcl-2-deficient mice. In addition, homozygous TG.AC mice were bred with FVB/N mice to generate hemizygous TG.AC mice on an FVB/N background to serve as a gene-dosage control. The F1 progeny consisted of FVB/N(v-Ha-ras+/ ):C57BI/6,129(bcl-2+/+),FVB/N(v-Ha-ra s+/-):C57BI/6,129(bcl-2+/-), and FVB/N(v-Ha ras+/-,bcl-2+/+). Ten-week-old mice were dosed twice weekly for 10 wk with acetone, 1.25 microg of 7,12-tetradecanoylphorbol-13-acetate (TPA), or 2.5 microg of TPA, and papillomas were counted weekly. Papillomas were analyzed for ras transgene and bcl-2 expression by reverse transcription-polymerase chain reaction, v-Ha-ras expression by in situ hybridization, and proliferating cell nuclear antigen expression by immunohistochemical analysis. Fewer papillomas (P < 0.05) were observed at the low dose of TPA (1.25 microg) in mice carrying the bcl 2 knockout allele than in the wild-type mice, suggesting that reduction of the bcl-2 gene product affects the susceptibility of TG.AC mice to TPA-induced papillomas. However, at the high dose of TPA (2.5 microg), there was no difference in papilloma response between knockout and wild-type mice, regardless of strain background. This suggests that at the higher dose of TPA, the effect of reduction in bcl-2 gene product was obscured. These results support the hypothesis that bcl-2 plays a limited role in skin tumorigenesis in the TG.AC mouse. PMID- 9328438 TI - Chromosomal and genetic alterations of 7,12-dimethylbenz[a]anthracene-induced melanoma from TP-ras transgenic mice. AB - The TP-ras transgenic mouse line expresses an activated human T24 Ha-ras gene with a mutation in codon 12, regulated by a mouse tyrosinase promoter. The transgene is expressed in melanocytes of the skin, eyes, and brain. The mice develop cutaneous melanoma when treated with 7,12-dimethylbenz[a]anthracene. Cell lines have been generated from the cutaneous tumors and metastatic lesions. By using fluorescence in situ hybridization with mouse whole chromosome paints, the cell lines were characterized for chromosomal abnormalities. Key findings in the tumor cells included translocations of chromosome 4 and alterations in chromosome 6. One tumor cell line contained a double translocation involving chromosomes 3 and 6. To extend the results of the chromosome 4 painting, Southern analysis of the p15INK4B, p16INK4A, and p19INK4D genes was performed. Our data indicated that there were homozygous and partial allelic deletions and polymorphisms in the region of chromosome 4 containing these genes, resulting in the absence or reduced expression of the p16 product. These findings are similar to those reported for human melanoma, and the TP-ras transgenic mouse may therefore be a valuable model for studying novel strategies for melanoma prevention and treatment. PMID- 9328439 TI - Identification of differentially expressed genes in chemically induced skin tumors. AB - Previous studies have demonstrated a role for the fos gene in promoting malignant conversion of mouse skin tumors. In the study reported here, differential display was performed to identify fos- and jun-regulated genes that are differentially expressed during premalignant progression. Total RNA isolated from variants of the papilloma cell line SP-1 transduced with retroviral vectors expressing v-jun and v-fos alone or in tandem was analyzed for the presence of differentially expressed transcripts by using 35 different primer combinations. Differentially expressed clones were rescreened by dot-blot analysis by using cDNA from chemically induced tumors with a high or low risk of malignant conversion. Three differentially displayed fragments were isolated in this analysis. Homology searches indicated that these fragments shared significant homology with the apoptosis inhibitor bcl-2, human alternative splicing factor/splicing factor 2 (ASF/SF2), and a novel gene not present in the GenBank or EMBL databases. In situ hybridization indicated that the expression levels of the bcl-2 homolog increased with malignant potential in chemically derived mouse skin tumors. A similar analysis indicated that expression of the ASF/SF2 homolog was greater in papillomas than in normal skin or in squamous cell carcinomas. Transcripts for this gene were most abundant in the granular layer. The expression pattern of the third differential display fragment was consistent with that of a tumor suppressor gene. This gene was expressed at very high levels in normal skin and benign papillomas but was essentially undetectable in squamous cell carcinomas. Through this approach, we identified known and novel genes that may contribute to malignant progression in epidermal tumors. PMID- 9328440 TI - Resistance of transformed mouse keratinocytes to growth inhibition by glucocorticoids. AB - Glucocorticoid hormones are strong inhibitors of normal keratinocyte proliferation, but established mouse skin papillomas and carcinomas become resistant to these hormones. The biological effect of glucocorticoids is mediated through a highly specific glucocorticoid receptor (GR). To study the possible mechanisms of glucocorticoid resistance of transformed mouse keratinocytes, we evaluated GR expression and function in non-tumorigenic (3PC), papilloma producing (MT1/2 and P1/17), and squamous cell carcinoma-producing (Ca3/7 and Ca8/29) keratinocyte cell lines and analyzed the DNA sequence of GR in glucocorticoid-sensitive and glucocorticoid-resistant keratinocytes. All transformed keratinocyte cell lines studied appeared to be completely resistant to the growth inhibition by the glucocorticoid fluocinolone acetonide (FA), whereas the untransformed cell line 3PC was very sensitive to FA. Despite the glucocorticoid resistance, all the tumorigenic keratinocyte cell lines expressed high levels of GR mRNA and protein. Southern blot analysis and direct sequencing of the DNA-binding domain of the GR gene revealed no significant changes in GR gene structure in transformed keratinocytes. To test the functional capability of GR, we compared the effect of FA on the expression of glucocorticoid-responsive genes. FA strongly induced metallothionein 1 expression in 3PC cells, slightly induced metallothionein 1 expression in P1/17 and Ca3/7 cells, and did not affect its expression in MT1/2 and Ca8/29 cells. These data suggest that resistance to the growth inhibition of glucocorticoids is an important feature of tumorigenic keratinocyte cell lines. It is likely that this hormone-resistant phenotype is a result of alteration of GR function but not of GR expression or gene structure. PMID- 9328441 TI - Kinetics of wound-induced v-Ha-ras transgene expression and papilloma development in transgenic Tg.AC mice. AB - The Tg.AC transgenic mouse, which harbors an activated v-Ha-ras coding region that is fused to an embryonic zeta globin transcriptional control region and a 3' simian virus 40 polyadenylation sequence, rapidly develops epidermal papillomas in response to topical application of chemical carcinogens or tumor promoters or to full-thickness wounding of the dorsal skin. In this report, we investigated the localization and temporal induction of v-Ha-ras transgene expression after full-thickness wounding of Tg.AC mouse skin. Surgically inflicted full-thickness incisions 3 cm long yielded four to six papillomas per Tg.AC mouse by 5 wk after wounding. Similar wounding of the FVB/N isogenic host strain did not produce tumors, which implicates a causal role for the v-Ha-ras transgene. Reverse transcription-polymerase chain reaction assays detected the v-Ha-ras transgene transcript in total RNA samples isolated from wound-associated tissue 3 and 4 wk after wounding. Tissues 1-2 wk after wounding and all non-wound-associated tissues were negative for transgene expression. In situ hybridization experiments using transgene-specific 35S-labeled antisense RNA probes localized transgene expression to the basal epidermal cells in wound-induced papillomas. Adjacent normal and hyperplastic skin tissues were negative for transgene expression by this assay. This work supports the hypothesis that the wound repair response leads to the transcriptional activation and continued expression of the v-Ha-ras transgene in specific cells in the skin, which alters normal epithelial differentiation and ultimately results in neoplastic growth. PMID- 9328442 TI - Evidence that mirex promotes a unique population of epidermal cells that cannot be distinguished by their mutant Ha-ras genotype. AB - Mirex is a potent tumor promoter in 7,1 2-dimethylbenz[a]anthracene (DMBA) initiated female CD-1 mouse skin. Like 12-O-tetradecanoylphorbol-13-acetate (TPA), mirex promotes papillomas that have a Ha-ras mutation; however, unlike TPA promotion, mirex promotion does not involve a general hyperplastic response. We used proliferating cell nuclear antigen (PCNA) and 5-bromo-2'-deoxyuridine (BrdU) immunohistochemical staining to further examine the proliferative capacity of mirex. The numbers of PCNA- and BrdU-positive epidermal S-phase cells were highly concordant in all treatment groups. Unlike a single application of TPA, a single application of mirex had little or no effect on the number of S-phase epidermal cells, and chronic application of mirex to mouse skin produced only minimal increases in S-phase cells. Moreover, mirex did not significantly alter the growth of BALB/MK-2 keratinocytes in media containing either 0.05 or 1.2 mM Ca++. These results suggest that mirex may have highly specific effects on the proliferation of initiated cells and support the existence of a unique mirex mechanism and/or distinct population of mirex-promotable mutant Ha-ras epidermal cells. To begin to address this issue of a distinct population of mirex promotable mutant Ha-ras cells, we conducted a tandem experiment in which DMBA initiated mice were treated twice weekly with a maximal promoting dose of mirex. Then, when the number of papillomas reached a plateau, these same mice were treated twice weekly with a maximal promoting dose of TPA. Mice treated with mirex developed a maximum of 6.4 papillomas/mouse. These mice were then promoted with TPA, which produced 8.9 additional papillomas/mouse for a total of 15.3 papillomas/mouse. The maximum tumor yields from other groups of mice treated with only TPA or mirex were 9.8 and 7.3 papillomas/mouse, respectively. Therefore, under these tandem conditions, tumor yields were additive, indicating that there are at least two distinct populations of mutant Ha-ras cells: one promoted by mirex and the other by TPA. PMID- 9328444 TI - Increased apoptosis during papilloma development in mice susceptible to tumor progression. AB - Programmed cell death (apoptosis) is known to occur not only during normal development and tissue remodeling but also during neoplasia. Despite the suggested role of apoptosis in preventing the proliferation of malignant cells, a positive correlation between tumor progression and the presence of apoptotic cells has been found in different types of cancer, including epithelial tumors. In normal mouse skin, the role of apoptosis is not completely understood, and it has been suggested that terminal differentiation may be a special case of apoptosis. In the work reported here, we counted apoptotic cells in mouse skin tumors generated with a two-stage chemical carcinogenesis protocol. We analyzed papillomas from outbred SENCAR mice at different times during promotion, and to better determine the correlation between apoptosis and tumor progression, we compared papillomas generated from two inbred strains derived from the SENCAR stock that differ in their susceptibility to tumor progression. Our results showed that in mouse skin chemical carcinogenesis, the number of apoptotic cells was greater in papillomas that may have been in the process of progressing to squamous cell carcinomas. This conclusion is also supported by the fact that papillomas from SENCAR P/Bt. mice, a tumor progression-susceptible strain derived from outbred SENCAR mice, had more apoptotic cells than papillomas from progression-resistant SSIN mice. PMID- 9328443 TI - Changes in protein expression during multistage mouse skin carcinogenesis. AB - To directly compare the expression patterns of different proteins known to be altered during mouse skin carcinogenesis, serial sections of normal and hyperplastic skin and tumors from various stages of 7,12 dimethylbenz[a]anthracene-initiated, 12-O-tetradecanoylphorbol-13-acetate promoted female SENCAR mice were examined by immunohistochemistry. In untreated, normal mouse skin, keratin 1 (K1) and transforming growth factor-beta1 (TGFbeta1) were strongly expressed in the suprabasal layers, whereas integrin alpha6beta4 was expressed only in basal cells and only moderate staining for transforming growth factor-alpha (TGFalpha) was seen. In hyperplastic skin, TGFalpha expression became stronger, whereas expression of another epidermal growth factor (EGF) receptor ligand, heparin-binding EGF-like growth factor (HB-EGF), was strongly induced in all epidermal layers from no expression in normal skin. Likewise, the gap-junctional protein connexin 26 (Cx26) became highly expressed in the differentiated granular layers of hyperplastic skin relative to undetectable expression in normal skin. Expression of cyclin D1 in the proliferative cell compartment was seen in all benign and malignant tumors but not in hyperplastic skin. Beginning with very early papillomas (after 10 wk of promotion), expression of alpha6beta4 in suprabasal cells and small, focal staining for keratin 13 (K13) were seen in some tumors. Later (after 20-30 wk), focal areas of gamma-glutamyl transpeptidase (GGT) activity appeared in a few papillomas, whereas TGFbeta1 expression began to decrease. Cx26 and TGFalpha staining became patchier in some late-stage papillomas (30-40 wk), whereas suprabasal alpha6beta4, K13, and GGT expression progressively increased and K1 expression decreased. Finally, in squamous cell carcinomas (SCCs), there was an almost complete loss of K1 and a further decline in TGFalpha, HB-EGF, TGFbeta1, and Cx26 expression. On the other hand, almost all SCCs showed suprabasal staining for alpha6beta4 and widespread cyclin D1 and K13 expression, whereas only about half showed positive focal staining for GGT activity. PMID- 9328445 TI - New strains of inbred SENCAR mice with increased susceptibility to induction of papillomas and squamous cell carcinomas in skin. AB - To develop mouse strains useful for studies of susceptibility and resistance to the induction of skin tumors, three new inbred SENCAR strains were independently derived by random inbreeding of outbred SENCAR mice. Characterization of these mice for sensitivity to skin tumor development indicated that mice of all three strains displayed increased sensitivity to initiation by 7,12 dimethylbenz[a]anthracene (DMBA), urethane, or N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and promotion by 12-O-tetradecanoylphorbol-13-acetate (TPA). Promotion by mezerein as well as carcinogenesis by repeated treatment with DMBA or MNNG produced papillomas with a high frequency of conversion to squamous cell carcinomas (SCCs). Compared with outbred SENCAR mice, development of both squamous papillomas and carcinomas was increased at least two-fold by all protocols tested. The F1 hybrid between SENCARA/Pt males and resistant BALB/cAnPt females was resistant to the induction of both papillomas and SCCs after initiation by 2 microg of DMBA and promotion by 20 weekly applications of 2 microg of TPA. Papillomas developed in all of the SENCARA/Pt mice, none of the BALB/cAnPt mice, and 12% of the F1 progeny. Thus, at these doses of initiator and promoter, resistance was incompletely dominant in the F1 hybrid. However, the responsiveness of the F1 mice could be increased substantially by increasing the dose of the promoter. PMID- 9328446 TI - Evidence that initiated keratinocytes clonally expand into multiple existing hair follicles during papilloma histogenesis in SENCAR mouse skin. AB - We have previously shown that the precursors of cutaneous papillomas in SENCAR mice initiated with 7,12-dimethylbenz[a]anthracene and promoted with 12-O tetradecanoylphorbol-13-acetate are focal hyperplastic lesions that we refer to as squamous cell hyperplastic foci (SCHF). Ha-ras gene codon 61 mutations were frequently found in SCHF, providing evidence that these lesions represent clones of initiated cells. We report here the pathogenesis of multiple hair follicle involvement in more advanced SCHF and describe the role of the hair follicle in papilloma histogenesis. Detailed histological evaluation of 83 SCHF and 25 early papillomas revealed a morphological continuum from the least developed SCHF, involving only one hair follicle, to advanced SCHF and early papillomas, which involved more than 10 hair follicles. These results provide evidence of the recruitment of additional hair follicles as SCHF progress. In advanced SCHF and early papillomas the bulk of the epithelial component in all cases consisted of several markedly hyperplastic adjacent hair follicles, whereas the involved interfollicular epidermis (IFE) was generally less hyperplastic. All of the hair follicles involved in SCHF appeared to have been preexisting, based on their pattern of spacing, that they were consistently normal appearing below the level of the sebaceous glands, and that they were in the same phase of the hair cycle as surrounding, uninvolved hair follicles. Also, no evidence of follicular neogenesis was observed in serially sectioned SCHF, and coalescence of smaller lesions was rare. To investigate whether the involvement of multiple hair follicles in SCHF was due to expansion of initiated cells into existing hair follicles or, possibly, to a paracrine mechanism, we analyzed different levels of three serially sectioned SCHF and one early papilloma for Ha-ras mutations. These analyses revealed cells with Ha-ras gene codon 61 mutations at multiple levels that involved different hair follicles. Overall, our results provide evidence that as initiated cells clonally expand, they spread across the IFE and populate the upper permanent portions of existing hair follicles. The abnormal proliferation of the infundibula of the hair follicles involved in SCHF appears to give rise to most of the epithelial component of papillomas. PMID- 9328447 TI - MNDA binds NPM/B23 and the NPM-MLF1 chimera generated by the t(3;5) associated with myelodysplastic syndrome and acute myeloid leukemia. AB - The myeloid cell nuclear differentiation antigen (MNDA) is a nuclear protein expressed specifically in developing cells of the human myelomonocytic lineage, including the end-stage monocytes/macrophages and granulocytes. Nuclear localization, lineage- and stage-specific expression, association with chromatin, and regulation by interferon alpha indicate that this protein is involved in regulating gene expression uniquely associated with the differentiation process and/or function of the monocyte/macrophage. MNDA does not bind specific DNA sequences, but rather a set of nuclear proteins that includes nucleolin (C23). Both in vitro binding assays and co-immunoprecipitation were used to demonstrate that MNDA also binds protein B23 (nucleophosmin/NPM). Three reciprocal chromosome translocations found in certain cases of leukemia/lymphoma involve fusions with the NPM/B23 gene, t(5;17) NPM-RARalpha, t(2;5) NPM-ALK, and the t(3;5) NPM-MLF1. In the current study, MNDA was not able to bind the NPM-ALK chimera originating from the t(2;5) and containing residues 1-117 of NPM. However, MNDA did bind the NPM-MLF1 product of the t(3;5) that contains the N-terminal 175 residues of NPM. The additional 58 amino acids (amino acids 117-175) of the NPM sequence that are contained in the product of the NPM-MLF1 fusion gene relative to the product of the NPM-ALK fusion appear responsible for MNDA binding. This additional NPM sequence contains a nuclear localization signal and clusters of acidic residues believed to bind nuclear localization signals of other proteins. Whereas NPM and nucleolin are primarily localized within the nucleolus, MNDA is distributed throughout the nucleus including the nucleolus, suggesting that additional interactions define overall MNDA localization. PMID- 9328448 TI - Dual activity of CSF-1, produced by a murine hybrid cell line and from other sources, on the growth of bone marrow stromal cells in culture. AB - Medium conditioned by the cell line A.4.10 was shown to produce a factor that inhibits the growth of bone marrow (BM) stromal foci in liquid cultures. In this study we demonstrated that CSF-1, present in this conditioned medium (CM), was largely responsible for this inhibition. This was concluded from the following findings: 1) the CSF-1 activity in the CM copurified with the inhibitory activity in several biochemical purification procedures; 2) an antibody against murine CSF 1 was able to neutralize the CSF-1 activity as well as the inhibitory activity in the A.4.10 CM; and 3) immunopurified murine CSF-1 from A.4.10 CM and from another murine source and recombinant human (rh) CSF-1 were all able to inhibit stromal cell growth. In secondary stromal cultures, inhibition was demonstrated at high cell densities but not at low densities, suggesting that accessory cells may be involved in the effect. When the dose rh CSF-1 was reduced to half of that required to produce 50% inhibition, stimulation of primary stromal foci growth was demonstrated, supporting observations by others that CSF-1 can act as a growth factor for BM stromal cells. In this report we thus demonstrate both inhibitory and stimulatory effects of CSF-1 on the growth of BM stromal cells in vitro. PMID- 9328449 TI - Ex vivo expansion of megakaryocyte progenitors: effect of various growth factor combinations on CD34+ progenitor cells from bone marrow and G-CSF-mobilized peripheral blood. AB - Prolonged thrombocytopenia resulting from inadequate megakaryocyte (MK) progenitor cell reconstitution is a serious complication of hematopoietic cell supported high-dose chemotherapy (HDC). In this situation, the infusion of MK progenitors that are expanded ex vivo could be clinically beneficial. In this study we investigated the ability of various growth factor combinations to generate MK progenitors. CD34+ cells derived from bone marrow (BM) and granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood (PB) from 17 patients with breast cancer, lymphoma, or myeloma were cultured unpertubed for 10 days in a serum-free liquid culture system that contained recombinant growth factors. Five different growth factors combinations were evaluated: Stem cell factor (SCF), interleukin (IL)-3, IL-6 + G-CSF (combination 1); SCF, megakaryocyte growth and development factor (MGDF) + G-CSF (combination 2); SCF + MGDF (combination 3); MGDF alone (combination 4); and SCF, IL-3, IL-6, G-CSF + MGDF (combination 5). PB CD34+ cells yielded significantly higher numbers of CD41+ MK progenitors than BM CD34+ cells with any of the growth factor regimens assayed. PB CD34+ cells (2x10[5]) at day 0 generated 1.2 to 1.3x10(6) CD41+ cells by day 10 when cultured in the presence of growth factor combinations 1, 2, or 3. In contrast, 2x10(5) BM CD34+ cells produced 5x10(5) CD41+ cells after 9 days in the presence of combination 1, whereas lower numbers of CD41+ cells were generated in cultures with combinations 2 and 3 (2.3x10[5] and 4.2x10[4], respectively). The addition of MGDF to cultures that were grown with combination 1 for 5 days increased the number of CD41+ cells (1.7-fold increase in PB-derived cultures, 1.6-fold increase in BM-derived cultures). Treatment with MGDF alone resulted in higher frequencies of MK progenitors than those obtained in cultures with combined growth factors (79% in PB-derived cultures, 25% in BM derived cultures), but because total cell growth was attenuated, absolute numbers of MK progenitors were lower (7x10(5) in PB-derived cultures, 7x10(4) in BM). Morphological analysis of immunocytochemically identified megakaryocytic cells revealed mononuclear cells as the predominant cell type in all of the cultures. During the 10-day culture period, PB-derived MK progenitors did not show notable maturation, even under the influence of MGDF, whereas in BM-derived cultures MGDF induced a significant shift to binuclear cells and stage I MK after day 5. Phenotypic analysis of cell surface markers showed that the majority of cultured megakaryocytic cells coexpressed CD34 and platelet glycoproteins (GPs), also indicating an immature stage of development. The ex vivo proliferative activity of CD34+ cells and their potential to develop into the megakaryocytic lineage demonstrated considerably high interpatient variations. There was no correlation between platelet recovery following HDC with hematopoietic cell support and the magnitude of GP+ cell expansion ex vivo, suggesting the feasibilty of MK expansion ex vivo in patients with prolonged thrombocytopenia posttransplantation. In summary, these data indicate that GCSF-mobilized CD34+ PBPCs are more effectively expanded ex vivo into the megakaryocytic lineage than are CD34+ BMPCs. CD34+/GP+ MK progenitors may be an appropiate cell population for transplantion as prophylaxis or treatment of prolonged thrombocytopenia. The efficacy of this procedure will be tested prospectively in a clinical trial. PMID- 9328450 TI - Endogenous IL-2 production by natural killer cells maintains cytotoxic and proliferative capacity following retroviral-mediated gene transfer. AB - Interleukin (IL)-2 therapy given at tolerable doses is insufficient to induce maximum activation of natural killer (NK) cells. We recently demonstrated that NK cells expanded in vivo can be maximally activated by short-term ex vivo incubation with 1000 U/mL IL-2. However, IL-2 withdrawal, which would occur with reinfusion, may lead to a rapid loss of cell viability and function. We hypothesized that retroviral transduction could provide an endogenous source of IL-2 to maintain NK function as measured by proliferation and cytotoxicity. Enriched NK cells were transduced with supernatants containing an MFG-based retrovirus designed to express murine IL-2 cDNA. Several supernatant transduction strategies were evaluated. NK cells were initially cultured in 1000 U/mL of huIL2 for 7-8 days, harvested, and replated prior to transduction (4 hours at 37degrees C); this proved insufficient to sustain NK proliferation or maintain cytotoxicity after exogenous human IL-2 (huIL-2) withdrawal. An alternative transduction procedure using phosphate-depleted medium, centrifugation, and transduction for 16 hours at 32degrees C was then evaluated. NK cells transduced under these conditions maintained significant NK proliferation in the absence of exogenous IL 2 compared with sham-transduced controls. Two consecutive daily transductions resulted in less proliferation, suggesting that several exposures to retroviral supernatant may inhibit subsequent NK proliferation. Cytotoxicity of the transduced NK cells against K562 and Raji was maintained under these conditions without exogenous IL-2. Sham-transduced NK cells produced 8.3+/-2.6 U/mL of murine IL-2 (muIL-2) by ELISA (background) after 7 days without exogenous IL-2. In contrast, 109+/-23 U/mL muIL-2 was produced by NK cells transduced with supernatant from the MFG/muIL-2 producer line. These experiments demonstrate that NK cells can be successfully transduced with retroviruses and induced to express sufficient IL-2 to maintain their proliferative and cytotoxic functions. Transduction of IL-2 genes into NK cells may offer advantages over exogenous IL-2 administration in maintaining maximum function for use in antitumor immunotherapy. PMID- 9328451 TI - Erythropoietin-receptor expression and function during the initiation of murine yolk sac erythropoiesis. AB - Although erythropoietin is necessary for definitive (fetal liver and bone marrow) erythropoiesis, the role of erythropoietin signaling in primitive (yolk sac) hematopoiesis has not been well defined. In situ hybridization studies have revealed that erythropoietin-receptor (EPOR) mRNA accumulation begins in mesoderm cell masses of the developing yolk sac of the neural plate stage embryo (E7.5) before the development of morphologically recognizable erythroblasts. EPOR mRNA is also present in yolk sac blood islands at early somite stages (E8.5). These findings suggest that EPOR functions during early stages of yolk sac erythropoiesis. We have used a serum-free murine yolk sac explant system (Palis et al., Blood 86:156, 1995) to investigate the initial differentiation of primitive erythroblasts from extraembryonic mesoderm cells. Exogenous erythropoietin increased both erythroblast numbers and betaH1-globin accumulation in yolk sac explants, suggesting that primary yolk sac erythroblasts are directly responsive to erythropoietin. An antisense oligodeoxynucleotide (ODN) experimental approach was used to examine the functional role of erythropoietin signaling during the initiation of yolk sac hematopoiesis in yolk sac explants. Antisense EPOR ODN produced a >50% reduction (p < 0.005) in the number of differentiating primitive erythroblasts, >95% reduction in betaH1-globin accumulation (p < 0.001), and a >50% reduction (p < 0.01) in the number of CFU-E and BFU-E compared with missense EPOR ODN-treated and untreated control explants. Antisense EPOR ODN also blocked the increase in primitive erythroblast number induced by exogenous erythropoietin. We conclude that erythropoietin/EPOR signaling is functionally active during the initial proliferation and differentiation of primary yolk sac erythroblasts. These results also suggest that other growth factor signaling cascades are active during the onset of mammalian erythropoiesis. PMID- 9328452 TI - Analysis of myeloid-associated genes in human hematopoietic progenitor cells. AB - The distribution of myeloid lineage-associated cytokine receptors and lysosomal proteins was analyzed in human CD34+ cord blood cell (CB) subsets at different stages of myeloid commitment by reverse-transcriptase polymerase chain reaction (RT-PCR). The highly specific granulomonocyte-associated lysosomal proteins myeloperoxidase (MPO) and lysozyme (LZ), as well as the transcription factor PU.1, were already detectable in the most immature CD34+Thy-1+ subset. Messenger RNA (mRNA) levels for the granulocyte-colony stimulating factor (G-CSF) receptor, granulocyte-macrophage (GM)-CSF receptor alpha subunit and tumor necrosis factor (TNF) receptors I (p55) and II (p75) were also detected in this subset in addition to c-kit and flt-3, receptors known to be expressed on progenitor cells. By contrast, the monocyte-macrophage colony stimulating factor (M-CSF) receptor was largely absent at this stage and in the CD34+Thy-1-CD45RA- subsets. The M-CSF receptor was first detectable in the myeloid-committed CD34+Thy-l-CD45RA+ subset. All other molecules studied were found to be expressed at this stage of differentiation. Different cocktails of the identified ligands were added to sorted CD34+Thy-1+ single cells. Low proliferative capacity was observed after 1 week in culture in the presence of stem cell factor (SCF) + Flt-3 ligand (FL) + G CSF. Addition of GM-CSF to this basic cocktail consistently increased the clonogenic capacity of single CD34+Thy-1+ cells, and this effect was further enhanced (up to 72.3 +/- 4.3% on day 7) by the inclusion of TNF-alpha. In conclusion, the presence of myeloid-associated growth factor receptor transcripts in CD34+ CB subsets does not discriminate the various stages of differentiation, with the exception of the M-CSF receptor. In addition, we show that TNF-alpha is a potent costimulatory factor of the very immature CD34+Thy-1+ CB subset. PMID- 9328453 TI - Use of hematopoietic cytokines to accelerate the recovery of the immune system in irradiated mice. AB - In our previous studies aimed at designing appropriate strategies to accelerate recovery of the immune system after irradiation, we found that the hematopoietic cytokine recombinant murine (rmu) interleukin (IL)-3 was able to induce differentiation and growth of thymocytes and splenic T and B lymphocytes in mice exposed to x-rays (200-500 cGy). The recovery, however, was complete at 7 days only after a dose of 200 cGy, whereas 2, 3, and 4 weeks were necessary to achieve full recovery after 300, 400, and 500 cGy, respectively. These studies were extended to investigate the effects of another hematopoietic cytokine, recombinant human (rhu) IL-11, a bone marrow stromal-derived cytokine, administered together with IL-3 to irradiated mice. The synergistic effect of the two cytokines was evident when relatively small doses of rhu IL-11 were injected with an optimal dose of rmu IL-3. PMID- 9328454 TI - Expression mapping of adhesion receptor genes during differentiation of individual hematopoietic precursors. AB - Associations between hematopoietic cells and their microenvironment are central to the development and maintenance of a functional hematopoietic system. It is important, therefore, to identify the surface receptors that mediate adhesive interactions among hematopoietic cells, stromal cells, and extracellular matrix components. In this study, we examined the expression of mRNA transcripts encoding a number of cell adhesion molecules and surface antigens in primitive hematopoietic cells isolated from murine bone marrow and fetal liver. Using a panel of probes, we hybridized a library of globally amplified cDNA prepared by reverse transcriptase-polymerase chain reaction of poly(A)+ mRNA from individual precursors and mature cell populations, representing precisely defined positions within the hematopoietic developmental hierarchy. The panel included probes specific for the CD45, CD34, P-glycoprotein (mdr1), Ly-6A/E (Sca-1), heat stable antigen (CD24), Fc receptor for IgG FcgammaRII (CD32), CD44, CD22, and ICAM-1 (CD54) genes, as well as alphaL (CD11a), alphaM (CD11b), beta2 (CD18), alpha4 (CD49d), alpha5 (CD49e), beta1 (CD29), and beta7 integrin subunit sequences. The data, which revealed stage- and lineage-specific expression patterns, should prove useful in designing future mechanistic studies aimed at elucidating the role played by adhesion receptors in normal and abnormal hematopoiesis. PMID- 9328455 TI - Vesnarinone exhibits antitumor effect against myeloid leukemia cells via apoptosis. AB - Vesnarinone is a positive inotropic agent used for treating congestive heart failure. We evaluated its ex vivo effects on myeloid leukemia cell lines and primary acute myelogenous leukemia cells. Vesnarinone inhibited the incorporation of radiolabeled thymidine by a myeloid cell line, HL60, in a dose-dependent manner at concentrations ranging from 0.1 to 30 microg/mL. A maximum 40% suppression was seen at a concentration of 10 microg/mL. Determination of viable cell counts by trypan blue dye exclusion method demonstrated vesnarinone to be cytocidal for HL60 cells. Vesnarinone induced DNA fragmentation as detected by electrophresis in HL60 cells after 72-hour culture; this effect was not inhibited by G-CSF. The apoptosis induced by vesnarinone was also detected by the in situ end-labeling method. Northern blot analysis showed a reduction of c-myc mRNA expression in HL60 cells by vesnarinone. However, immunostaining assay showed no change in the expression of Fas and Bcl-2 proteins. We next examined the effect of vesnarinone on primary myeloid leukemia cells derived from 10 patients: 3 cases of M1, 2 of M2, 3 of M3, 1 of M4, and 1 of M6, by the French-American British classification. Vesnarinone inhibited the incorporation of thymidine in all cells, with a mean suppression of 58.1%. DNA electrophoresis showed induction of DNA fragmentation in cultured cells with vesnarinone for 72 hours in 8 of the 10 patients with primary leukemia. However, bone marrow mononuclear cells from healthy controls showed no growth suppression or DNA fragmentation in response to vesnarinone. These results suggest that vesnarinone may be useful in treating myeloid leukemia. PMID- 9328456 TI - Severe hypoxia enhances the formation of erythroid bursts from human cord blood cells and the maintenance of BFU-E in vitro. AB - Incubation in severe hypoxia (1% oxygen) increased the number of erythroid bursts generated from full-term CD34+, or premature mononucleated, human cord blood (CB) cells, in semisolid cultures containing stem cell factor (SCF), interleukin (IL) 3 and erythropoietin (EPO). Severe hypoxia also enhanced the maintenance of erythroid burst-forming units (BFU-E) in CB cell liquid cultures. These positive effects of hypoxia on the maintenance and cloning efficiency of BFU-E did not extend to the other progenitors assayed. Hypoxia, on the other hand, markedly reduced the size and level of hemoglobinization of bursts and, in liquid cultures, suppressed the growth factor-stimulated numerical increase in BFU-E and inhibited the expression of CD36, a marker of erythroid colony-forming units and maturing erythroid precursors. However, when transferred to clonal assays incubated in air, cells from liquid cultures incubated in hypoxia or in air generated fully expanded and hemoglobinized bursts, suggesting that in hypoxia the clonogenic potential of BFU-E was maintained and the development of erythroid clones reversibly inhibited. These results indicate that hypoxia inversely regulates two subsequent phases of erythropoiesis, i.e., it enhances the maintenance of BFU-E and the early development of erythroid clones but inhibits the terminal expansion and maturation of these clones. The cloning of CB cells selected for CD34 positivity, when compared with that of the total population of mononucleated CB cells, revealed that the early development of erythroid bursts was either hypoxia-enhanced or hypoxia-insensitive, reflecting the existence of two different types of BFU-E. Hypoxia-enhanced BFU-E are relatively immature, are maintained in hypoxia but not in air, and account for a large part of CD34+ BFU-E and for a high percentage of the BFU-E in premature CB. Hypoxia-insensitive BFU-E are mostly CD34- and are largely predominant in full-term CB, and most probably correspond to a more mature type of BFU-E. PMID- 9328457 TI - Clinical and managed care issues in blood and marrow transplantation for hematologic diseases. Report of a symposium, 14 March 1996, Washington, DC. PMID- 9328462 TI - Mammalian telomerase: catalytic subunit and knockout mice. AB - For the second time this year random cDNA sequencing, in combination with data from unicellular eukaryotes, has made a significant contribution to the analysis of human telomerase. Two groups have reported mammalian homologues of the Tetrahymena p80 telomerase-associated protein, in both cases the key breakthrough being mammalian cDNA clones with database matches to Tetrahymena p80. This has now been joined by the sequence of a candidate for the human telomerase catalytic subunit. The discovery that its message abundance closely follows telomerase activity could make a major impact on the utility of telomerase as a diagnostic marker for human malignancy. In addition, Blasco et al . report the phenotype of a transgenic mouse deleted for the mTR gene, which encodes the essential RNA component of telomerase. Interestingly tumour formation is unaffected in these mice, strengthening the argument that telomerase expression in mouse tumourigenesis is an innocent bystander rather than a necessary event. However, fundamental differences between the genomic organisation of mouse and human telomeres mean that the mouse is not a straightforward model to critically test the role of telomere loss and telomerase in human malignancy. PMID- 9328463 TI - Rethinking genotype and phenotype correlations in polyglutamine expansion disorders. PMID- 9328464 TI - Isolation of a candidate human telomerase catalytic subunit gene, which reveals complex splicing patterns in different cell types. AB - Telomerase is a multicomponent reverse transcriptase enzyme that adds DNA repeats to the ends of chromosomes using its RNA component as a template for synthesis. Telomerase activity is detected in the germline as well as the majority of tumors and immortal cell lines, and at low levels in several types of normal cells. We have cloned a human gene homologous to a protein from Saccharomyces cerevisiae and Euplotes aediculatus that has reverse transcriptase motifs and is thought to be the catalytic subunit of telomerase in those species. This gene is present in the human genome as a single copy sequence with a dominant transcript of approximately 4 kb in a human colon cancer cell line, LIM1215. The cDNA sequence was determined using clones from a LIM1215 cDNA library and by RT-PCR, cRACE and 3'RACE on mRNA from the same source. We show that the gene is expressed in several normal tissues, telomerase-positive post-crisis (immortal) cell lines and various tumors but is not expressed in the majority of normal tissues analyzed, pre-crisis (non-immortal) cells and telomerase-negative immortal (ALT) cell lines. Multiple products were identified by RT-PCR using primers within the reverse transcriptase domain. Sequencing of these products suggests that they arise by alternative splicing. Strikingly, various tumors, cell lines and even normal tissues (colonic crypt and testis) showed considerable differences in the splicing patterns. Alternative splicing of the telomerase catalytic subunit transcript may be important for the regulation of telomerase activity and may give rise to proteins with different biochemical functions. PMID- 9328465 TI - The IPL gene on chromosome 11p15.5 is imprinted in humans and mice and is similar to TDAG51, implicated in Fas expression and apoptosis. AB - We searched for novel imprinted genes in a region of human chromosome 11p15.5, which contains several known imprinted genes. Here we describe the cloning and characterization of the IPL ( I mprinted in P lacenta and L iver) gene, which shows tissue-specific expression and functional imprinting, with the maternal allele active and the paternal allele relatively inactive, in many human and mouse tissues. Human IPL is highly expressed in placenta and shows low but detectable expression in fetal and adult liver and lung. Mouse Ipl maps to the region of chromosome 7 which is syntenic with human 11p15.5 and this gene is expressed in placenta and at higher levels in extraembryonic membranes (yolk sac), fetal liver and adult kidney. Mouse and human IPL show sequence similarity to TDAG51 , a gene which was shown to be essential for Fas expression and susceptibility to apoptosis in a T lymphocyte cell line. Like several other imprinted genes, mouse and human IPL genes are small and contain small introns. These data expand the repertoire of known imprinted genes and will be helpful in testing the mechanism of genomic imprinting and the role of imprinted genes in growth regulation. PMID- 9328467 TI - The human GARS-AIRS-GART gene encodes two proteins which are differentially expressed during human brain development and temporally overexpressed in cerebellum of individuals with Down syndrome. AB - Purines are critical for energy metabolism, cell signalling and cell reproduction. Nevertheless, little is known about the regulation of this essential biochemical pathway during mammalian development. In humans, the second, third and fifth steps of de novo purine biosynthesis are catalyzed by a trifunctional protein with glycinamide ribonucleotide synthetase (GARS), aminoimidazole ribonucleotide synthetase (AIRS) and glycinamide ribonucleotide formyltransferase (GART) enzymatic activities. The gene encoding this trifunctional protein is located on chromosome 21. The enzyme catalyzing the intervening fourth step of de novo purine biosynthesis, phosphoribosylformylglycineamide amidotransferase (FGARAT), is encoded by a separate gene on chromosome 17. To investigate the regulation of these proteins, we have generated monoclonal and/or polyclonal antibodies specific to each of these enzymatic domains. Using these antibodies on western blots of Chinese hamster ovary (CHO) cells transfected with the human GARS-AIRS-GART gene, we show that this gene encodes not only the trifunctional protein of 110 kDa, but also a monofunctional GARS protein of 50 kDa. This carboxy-truncated human GARS protein is produced by alternative splicing resulting in the use of a polyadenylation site in the intron between the terminal GARS and the first AIRS exons. The expression of both the GARS and GARS-AIRS-GART proteins are regulated during development of the human cerebellum, while the expression of FGARAT appears to be constitutive. All three proteins are expressed at high levels during normal prenatal cerebellum development while the GARS and GARS-AIRS-GART proteins become undetectable in this tissue shortly after birth. In contrast, the GARS and GARS AIRS-GART proteins continue to be expressed during the postnatal development of the cerebellum in individuals with Down syndrome. PMID- 9328469 TI - Germline mutations of the CDKN2 gene in UK melanoma families. AB - Germline mutations in CDKN2 on chromosome 9p21, which codes for the cyclin D kinase inhibitor p16, and more rarely, mutations in the gene coding for CDK4, the protein to which p16 binds, underlie susceptibility in some melanoma families. We have sequenced all exons of CDKN2 and analysed the CDK4 gene for mutations in 27 UK families showing evidence of predisposition to melanoma. Five different germline mutations in CDKN2 were found in six families. Three of the mutations (Met53Ile, Arg24Pro and 23ins24) have been reported previously. We have identified two novel CDKN2 mutations (88delG and Ala118Thr) which are likely to be associated with the development of melanoma, because of their co-segregation with the disease and their likely functional effect on the CDKN2 protein. In binding assays the protein expressed from the previously described mutation, Met53Ile, did not bind to CDK4/CDK6, confirming its role as a causal mutation in the development of melanoma. Ala118Thr appeared to be functional in this assay. Arg24Pro appeared to bind to CDK6, but not to CDK4. No mutations were detected in exon 2 of CDK4, suggesting that causal mutations in this gene are uncommon. The penetrance of these mutant CDKN2 genes is not yet established, nor is the risk of non-melanoma cancer to gene carriers. PMID- 9328468 TI - Structural and functional characterization of the human FMR1 promoter reveals similarities with the hnRNP-A2 promoter region. AB - Fragile X mental retardation syndrome is associated with an expansion of a CGG repeat within the 5'UTR of the first exon of the FMR1 gene, abnormal methylation of the CpG island in the promoter region, and a transcriptional silencing of this gene. We studied transcriptional regulation of the FMR1 gene using protein footprint analysis of the active and inactive gene in vivo . We identified four footprints within the FMR1 promoter region which correspond to consensus binding sites of known transcription factors, alpha-PAL/NRF1, Sp1, H4TF1/Sp1-like and c myc. These footprints were present in normal cells with a transcriptionally active FMR1 gene. The same footprints were present in different cell types: primary fibroblasts, lymphoblastoid cells and peripheral lymphocytes. However, for the 1.1 kb region analyzed, no footprints were detected in a variety of cell types derived from patients with fragile X syndrome which have a transcriptionally inactive FMR1 gene. A BLAST nucleotide search identified sequence similarities between the region of the FMR1 gene containing the footprints and an analogous region within the promoter region of the gene for the heterogeneous nuclear ribonucleoprotein (hnRNP) A2, a member of a family of ribonucleoproteins implicated in mRNA processing and nuclear-cytoplasm transport. The nucleotide sequences identified in the hnRNP-A2 promoter region correspond to the same consensus binding sites showing DNA-protein interactions in the FMR1 gene. Our previous functional studies and the studies of others demonstrate that FMR proteins, like hnRNP-A2, are also ribonucleoproteins which appear to be involved in mRNA transport. The results from our footprint studies suggest that the expression of the FMR1 gene is regulated by the binding of specific transcription factors to sequence elements in the 5' region of the gene and that this expression may be regulated by elements in common with the hnRNP-A2 gene. Common regulation of these two genes might play an important role in the cooperative processing and transport of mRNA from the nucleus to the translation machinery. PMID- 9328470 TI - Genetic control of serum IgE levels and asthma: linkage and linkage disequilibrium studies in an isolated population. AB - Immunoglobulin E (IgE) concentration in serum is elevated in atopic diseases such as asthma. A large genomic region on chromosome 5 has previously been implicated in the control of IgE levels and bronchial hyperreactivity and may, therefore, harbor genes predisposing to asthma. In an effort to confirm this linkage and to delimit the critical region, we took advantage of an isolated founder subpopulation in Finland to study genetic linkage and haplotype associations. Sixteen polymorphic markers, including the Interleukin-4 and -9 genes (IL4, IL9), were physically ordered and genotyped in 157 nuclear families. Genetic linkage studies involving sib- and cousin-pair analyses found no evidence of genetic linkage between markers in 5q and either serum IgE levels or asthma. Haplotype association studies were also performed. Although initial inspection suggested the possibility of linkage disequilibrium in the region of IL9, we developed a rigorous permutation test for assessing association and determined that the association was no greater than would be expected by chance. Sequence analysis of the IL9 gene in three patients sharing a possibly conserved haplotype revealed a T113M coding polymorphism, but this variant showed no association with either serum IgE levels or asthma. We conclude that allelic variation at chromosome 5q31 is not likely to contribute to inheritance of serum IgE levels or the development of asthma in this Finnish subpopulation. PMID- 9328471 TI - Genetic susceptibility for human familial essential hypertension in a region of homology with blood pressure linkage on rat chromosome 10. AB - Hypertension is a significant risk factor for heart attack and stroke and represents a major public health burden because of its high prevalence (e.g. 15 20% of the European and American populations). Although blood pressure is known to have a strong genetic determination, the genes responsible for susceptibility to essential hypertension are mostly unknown. Loci involved in blood pressure regulation have been found by linkage in experimental hereditary hypertensive rat strains, but their relationship to human hypertension has not been extensively investigated. One of the principal blood pressure loci has been mapped to rat chromosome 10 and we have undertaken an investigation of the homologous region on human chromosome 17 in familial essential hypertension. Affected sib-pair analysis and parametric analysis with ascertainment correction gave significant evidence of linkage ( P <0.0001 in some analyses) near two closely linked microsatellite markers, D17S183 and D17S934, that reside 18 cM proximal to the ACE locus in the homology region. Our results indicate that chromosome 17q could contain a susceptibility locus for human hypertension and show that comparative mapping may be a useful approach for identification of such loci in humans. PMID- 9328472 TI - A new pathogenic mutation in the APP gene (I716V) increases the relative proportion of A beta 42(43). AB - We report a novel mutation in the amyloid precursor protein gene (APP I716V) which probably leads to familial early onset Alzheimer's disease with an onset age in the mid 50s. Cells transfected with cDNAs bearing this mutation produce more A beta 1-42(43) than those transfected with wild-type APP and this effect is additive with that of the previously reported APP V717I mutation thus providing a novel approach for further increasing A beta 1-42(43) in model systems. PMID- 9328473 TI - Recurrent mutations in a single exon encoding the evolutionarily conserved olfactomedin-homology domain of TIGR in familial open-angle glaucoma. AB - Primary open-angle glaucoma (POAG) is a highly prevalent cause of irreversible blindness which associates cupping of the optic disc and alteration of the visual field, elevation of intraocular pressure being a major risk factor. Provided diagnosis is made at an early stage, treatments are available to prevent visual impairment. A locus, GLC1A, has been mapped on chromosome 1q23-q25 in several families affected with juvenile-onset POAG (JOAG) and also in some families affected with juvenile and middle-age onset POAG. Recently, three mutations of the TIGR (Trabecular meshwork-Induced Glucocorticoid Response) gene were shown to be responsible for the disease in several American families and in unrelated POAG patients. We now describe five new mutations in eight French families. All mutations known to date appear to concentrate in the evolutionarily conserved C terminal domain of TIGR which bears homology to frog olfactomedin, an extracellular matrix glycoprotein of the olfactory epithelium, to rat and human neuronal olfactomedin-related proteins and to F11C3.2, a protein from Caenorhabditis elegans . Moreover, this conserved domain of TIGR is encoded by a single exon to which mutation screening could be limited. Surprisingly, the TIGR message, which is abundantly transcribed in the trabecular meshwork and also in the ciliary body and the sclera, is not expressed in the optic nerve whose degeneration is, however, the primary lesion of POAG. PMID- 9328474 TI - Higher proportion of intact exon 9 CFTR mRNA in nasal epithelium compared with vas deferens. AB - The 5-thymidine (5T) variant of the cystic fibrosis transmembrane conductance regulator (CFTR) intron 8 polypyrimidine tract (IVS8-T tract) is the most frequent CFTR gene alteration identified in men with congenital bilateral absence of vas deferens (CBAVD). This alternative splicing variant gives rise to two transcripts, one normal with exon 9 intact and the other with in-frame deletion of exon 9. That CBAVD men usually have none of the other clinical signs of classical cystic fibrosis (CF) suggests less functional CFTR is produced in the reproductive tract than in other CF-associated organs. Nasal epithelia and segments of vas deferens were obtained from healthy, previously vasectomized men who presented for vasectomy reversal. Quantitative RT-PCR was performed on these specimens, with the region of CFTR cDNA spanning exon 9 amplified. For both nasal and vasal tissues, a strong positive correlation was found between the length of the IVS8-T tract and the proportion of mRNA with exon 9 intact. In addition, within the same subject, a significantly higher level of transcripts lacking exon 9 was found in vas deferens than nasal epithelia, regardless of the IVS8-T genotype. These findings suggest that the splicing of CFTR precursor mRNA is less efficient in vasal epithelia compared with respiratory epithelia. Thus, differential splicing efficiency between the various tissues which express CFTR provides one possible explanation for the reproductive tract abnormalities observed in infertile men with CFTR gene alterations but without other clinical manifestations of CF. PMID- 9328475 TI - Isolation of a new homeobox gene belonging to the Pitx/Rieg family: expression during lens development and mapping to the aphakia region on mouse chromosome 19. AB - We recently reported the positional cloning of a homeobox gene involved in the pathogenesis of Rieger syndrome, RIEG1 , and its mouse homolog, Rieg1 . Rieg1 (also independently described as Pitx2) is highly homologous to the Ptx1/Potx gene product, suggesting that there may be additional members of this novel Pitx family. The Pitx genes play an important role in eye, tooth, pituitary and umbilical region development as evidenced by Rieger syndrome and iris hypoplasia phenotypes, resulting from mutations in the RIEG1 gene and by expression studies. In order to characterize further the Pitx gene family we searched mouse cDNA libraries to identify additional members. A new gene was isolated which encodes a homeoprotein with strong homology to the other Pitx proteins and 97-100% identity in the homeodomain itself, suggesting that this is a third member of the family, Pitx3 . In whole mount in situ hybridization on mouse embryos ranging from 8.5 to 11.5 days post-coitum (d.p.c.), Pitx3 mRNA was seen only in the developing lens starting at day 11. Hybridization on cross-sections revealed strong signals in the lens vesicle in 11 d.p.c. embryos and throughout the lens, particularly in the anterior epithelium and equator region in 15 d.p.c. embryos. Pitx3 was mapped close to aphakia on mouse chromosome 19. The aphakia homozygous mouse is characterized by small eyes lacking a lens, which fail to develop beyond 11 d.p.c. These data make Pitx3 a strong candidate gene for the aphakia phenotype in the mouse and suggest a role for the human homolog in congenital lens malformations. PMID- 9328476 TI - Identification and characterization of human genes encoding Hprp3p and Hprp4p, interacting components of the spliceosome. AB - Nuclear RNA splicing occurs in an RNA-protein complex, termed the spliceosome. U4/U6 snRNP is one of four essential small nuclear ribonucleoprotein (snRNP) particles (U1, U2, U5 and U4/U6) present in the spliceosome. U4/U6 snRNP contains two snRNAs (U4 and U6) and a number of proteins. We report here the identification and characterization of two human genes encoding U4/U6-associated splicing factors, Hprp3p and Hprp4p, respectively. Hprp3p is a 77 kDa protein, which is homologous to the Saccharomyces cerevisiae splicing factor Prp3p. Amino acid sequence analysis revealed two putative homologues in Caenorhabditis elegans and Schizosaccharomyces pombe. Polyclonal antibodies against Hprp3p were generated with His-tagged Hprp3p over-produced in Escherichia coli . This splicing factor can co-immunoprecipitate with U4, U6 and U5 snRNAs, suggesting that it is present in the U4/U6.U5 tri-snRNP. Hprp4p is a 58 kDa protein homologous to yeast splicing factor Prp4p. Like yeast Prp4p, the human homologue contains repeats homologous to the beta-subunit of G-proteins. These repeats are called WD repeats because there is a highly conserved dipeptide of tryptophan and aspartic acid present at the end of each repeat. The primary amino acid sequence homology between human Hprp4p and yeast Prp4p led to the discovery of two additional WD repeats in yeast Prp4p. Structural homology between these human and yeast splicing factors and the beta-subunit of G-proteins has been identified by sequence-similarity comparison and analysis of the protein folding by threading. Structural models of Hprp4p and Prp4p with a seven-blade beta-propeller topology have been generated based on the structure of beta-transducin. Hprp3p and Hprp4p have been shown to interact with each other and the first 100 amino acids of Hprp3p are not essential for this interaction. These experiments suggest that both Hprp3p and Hprp4p are components of human spliceosomes. PMID- 9328477 TI - Evidence for uniparental, paternal expression of the human GABAA receptor subunit genes, using microcell-mediated chromosome transfer. AB - We have constructed mouse A9 hybrids containing a single normal human chromosome 15, via microcell-mediated chromosome transfer. Cytogenetic and DNA-polymorphic analyses identified mouse A9 hybrids that contained either a paternal or maternal human chromosome 15. Paternal specific expression of the known imprinted genes SNRPN (small nuclear ribonucleoprotein-associated polypeptide N gene) and IPW (imprinted gene in the Prader-Willi syndrome region) was maintained in the A9 hybrids. Using this system, we first demonstrated that human GABAAreceptor subunit genes, GABRB3 , GABRA5 and GABRG3 , were expressed exclusively from the paternal allele and that E6-AP (E6-associated protein or UBE3A ) was biallelically expressed. Moreover, the 5' portion of the GABRB3 gene was found to be hypermethylated on the paternal allele. Our data imply that GABAAreceptor subunit genes are imprinted and are possible candidates for Prader-Willi syndrome, and that this human monochromosomal hybrid system enables the efficient analysis of imprinted loci. PMID- 9328478 TI - Increased trinucleotide repeat instability with advanced maternal age. AB - Nucleotide repeat instability is associated with an increasing number of cancers and neurological disorders. The mechanisms that govern repeat instability in these biological disorders are not well understood. To examine genetic aspects of repeat instability we have introduced an expanded CAG trinucleotide repeat into transgenic mice. We have detected intergenerational CAG repeat instability in transgenic mice only when the transgene was maternally transmitted. These intergenerational instabilities increased in frequency and magnitude as the transgenic mother aged. Furthermore, triplet repeat variations were detected in unfertilized oocytes and were comparable with those in the offspring. These data show that maternal repeat instability in the transgenic mice occurs after meiotic DNA replication and prior to oocyte fertilization. Thus, these findings demonstrate that advanced maternal age is an important factor for instability of nucleotide repeats in mammalian DNA. PMID- 9328479 TI - Aberrant processing of the Fugu HD (FrHD) mRNA in mouse cells and in transgenic mice. AB - The puffer fish ( Fugu rubripes ) has a compact genome of 400 Mbp which is approximately 7.5-fold smaller than the human genome. It contains a similar number of genes but is deficient in intergenic, intronic and dispersed repetitive sequences. Fugu is becoming established as the model vertebrate genome for the identification and characterisation of novel human genes and conserved regulatory sequences. It has also been proposed that Fugu genes may provide natural mini genes for the production of transgenic mice. We have used the Fugu homologue of the Huntington's disease (HD) gene to test this possibility. The human and Fugu HD genes cover 170 kb and 23 kb respectively and have previously been sequenced in their entirety. In Fugu tissue, the Fugu HD gene was found to be expressed as predicted from the gene sequence but three differentially spliced forms were also detected. Despite the absence of conserved promoter sequences, the Fugu promoter was found to be functional in mouse cells. We have generated mice transgenic for the Fugu HD gene and conducted a detailed expression analysis across the entire 10 kb transcript. This revealed the presence of many aberrant splice forms which would be incompatible with the production of the Fugu huntingtin protein. The Fugu HD gene is incorrectly processed in mouse cells both in vitro and in vivo which sheds doubt on the usefulness of Fugu genes for transgenesis. PMID- 9328480 TI - Distortion of allelic expression of apolipoprotein E in Alzheimer's disease. AB - The APOE epsilon4 allele is a strong genetic susceptibility factor for Alzheimer's disease. Interaction with other biological factors may modulate the effect of the apoE isoforms. However, previous work suggested that other genetic variability within the APOE locus, influencing the effect of the epsilon4 allele, may exist. Such variability could modify the expression of the APOE gene and, in particular, the level of expression of APOE alleles could be an important determinant of disease pathogenesis. To test this hypothesis we examined the levels of expression of APOE in heterozygotes with AD and in controls, using a new method of semi-quantitation. We report that relative epsilon4 mRNA expression is increased in AD compared with controls and suggest that genetic variability in the neural expression of APOE contributes to disease risk. PMID- 9328481 TI - Molecular genetic and phenotypic analysis reveals differences between TSC1 and TSC2 associated familial and sporadic tuberous sclerosis. AB - Tuberous sclerosis (TSC) is an autosomal dominant disorder characterised by the development of hamartomatous growths in many organs. Sixty to seventy percent of cases are sporadic and appear to represent new mutations. TSC exhibits locus heterogeneity: the TSC2 gene is located at 16p13.3 whilst the TSC1 gene, predicted to encode a novel protein termed hamartin, has recently been cloned from 9q34. With the exception of a contiguous gene deletion syndrome involving TSC2 and PKD1 , TSC1 and TSC2 phenotypes have been considered identical. We have now comprehensively defined the TSC1 mutational spectrum in 171 sequentially ascertained, unrelated TSC patients by single strand conformation polymorphism and heteroduplex analysis of all 21 coding exons, and by assaying a restriction fragment spanning the whole locus. Mutations were identified in 9/24 familial cases, but in only 13/147 sporadic cases. In contrast, a limited screen revealed TSC2 mutations in two of the familial cases and in 45 of the sporadic cases. Thus TSC1 mutations were significantly under-represented among sporadic cases (Fisher's exact p -value = 3.12 x 10(-4)). Both large deletions and missense mutations were common at the TSC2 locus whereas most TSC1 mutations were small truncating lesions. Mental retardation was significantly less frequent among carriers of TSC1 than TSC2 mutations (odds ratio 5.54 for sporadic cases only, 6.78 +/- 1.54 when a single randomly selected patient per multigeneration family was also included). No correlation between mental retardation and the type of mutation was found. We conclude that there is a reduced risk of mental retardation in TSC1 as opposed to TSC2 disease and that consequent ascertainment bias, at least in part, explains the relative paucity of TSC1 mutations in sporadic TSC. PMID- 9328482 TI - Two different connexin 26 mutations in an inbred kindred segregating non syndromic recessive deafness: implications for genetic studies in isolated populations. AB - Non-syndromic recessive deafness (NSRD) is the most common form of prelingual hereditary hearing loss. To date, 10 autosomal NSRD loci (DFNBs) have been identified by genetic mapping; at least three times as many additional loci are expected to be identified. We have performed linkage analyses in two inter related inbred kindreds, comprised of >50 affecteds, from a single Israeli-Arab village segregating NSRD. Genetic mapping by two-point and multi-point linkage analysis in 10 candidate regions identified the segregating gene to be on human chromosome 13q11 (DFNB1). Haplotype analysis, using eight microsatellite markers spanning 15 cM in 13q11, suggested the segregation of two different mutations in this kindred: affected individuals were homozygotes for either haplotype or compound heterozygotes. The gene for the connexin 26 gap junction protein, recently shown to be mutant in both dominant and recessive deafness, maps to this locus. We identified two distinct mutations, W77R and Gdel35, both of which likely inactivate connexin 26. The Gdel35 change likely occurs at a mutational hotspot within the connexin 26 gene. The recombination of marker alleles at the polymorphisms studied in 13q11, at known map distances from the mutations, allowed us to estimate the age of the mutations to be 3-5 generations (75-125 years). This study independently confirms the identity of connexin 26 as an NSRD gene. Importantly, we demonstrate that in small populations with high rates of consanguinity, as compared with large outbred populations, recessive mutations may have very recent origin and show allelic diversity. PMID- 9328483 TI - IsK and KvLQT1: mutation in either of the two subunits of the slow component of the delayed rectifier potassium channel can cause Jervell and Lange-Nielsen syndrome. AB - The Jervell and Lange-Nielsen syndrome (JLNS) comprises profound congenital sensorineural deafness associated with syncopal episodes. These are caused by ventricular arrhythmias secondary to abnormal repolarisation, manifested by a prolonged QT interval on the electrocardiogram. Recently, in families with JLNS, Neyroud et al. reported homozygosity for a single mutation in KVLQT1 , a gene which has previously been shown to be mutated in families with dominantly inherited isolated long QT syndrome [Neyroud et al . (1997) Nature Genet ., 15, 186-189]. We have analysed a group of families with JLNS and shown that the majority are consistent with mutation at this locus: five families of differing ethnic backgrounds were homozygous by descent for markers close to the KVLQT1 gene and a further three families from the same geographical region were shown to be homozygous for a common haplotype and to have the same homozygous mutation of the KVLQT1 gene. However, analysis of a single small consanguineous family excluded linkage to the KVLQT1 gene, establishing genetic heterogeneity in JLNS. The affected children in this family were homozygous by descent for markers on chromosome 21, in a region containing the gene IsK . This codes for a transmembrane protein known to associate with KVLQT1 to form the slow component of the delayed rectifier potassium channel. Sequencing of the affected boys showed a homozygous mutation, demonstrating that mutation in the IsK gene may be a rare cause of JLNS and that an indistinguishable phenotype can arise from mutations in either of the two interacting molecules. PMID- 9328484 TI - Clinical advances in degenerative dementias. PMID- 9328485 TI - Comorbidity of mental disorders with substance misuse. PMID- 9328486 TI - General psychiatry in no-man's land. PMID- 9328487 TI - History of violent behaviour and schizophrenia in different cultures. Analyses based on the WHO study on Determinants of Outcome of Severe Mental Disorders. AB - BACKGROUND: Information on patterns and correlates of the violent behaviour of individuals with schizophrenia is largely limited to populations in developed countries. Data from a World Health Organization epidemiological study of schizophrenia and related disorders, the Determinants of Outcome of Severe Mental Disorders (DOSMD), presented an opportunity to study patterns of violence across multinational settings. METHOD: Centres in 10 countries participated in the DOSMD study. An incidence sample of 1017 patients with schizophrenia who had their first-in-lifetime contact with a helping agency as a result of their psychotic symptoms was obtained. Data were available on their history of violent behaviour, substance use, and demographics. RESULTS: The occurrence rate of assault in the entire cohort was 20.6 per hundred, but the rate was three times higher in the developing countries (31.5 per hundred) than in developed countries (10.5 per hundred). History of assault was associated with positive symptoms, such as excitement and auditory hallucinations, and with serious alcohol problems. CONCLUSIONS: The cultural context and the specific characteristics of the disease in individuals with schizophrenia may interactively affect rates of violent behaviour. PMID- 9328488 TI - Social networks and service use among representative cases of psychosis in south London. AB - BACKGROUND: Large social networks in patients with severe mental illness have been reported to be associated with a low rate of hospitalisation. We aim to determine whether social network size is related to the likelihood of hospitalisation and the amount of service use. METHOD: As part of a prospective controlled study, baseline interview data for a random sample of one-year prevalent cases with non-organic psychosis were analysed with respect to social network characteristics and service use during a six-month period. RESULTS: The likelihood of hospitalisation decreased with an increase in network size, while the number of services used by patients grew as the social network size increased. CONCLUSIONS: While larger social networks may be associated with a lower likelihood of hospitalization, they may also be related to wider use of non hospital services. PMID- 9328489 TI - Subjective experience of persistent schizophrenia and depression among US Latinos and Euro-Americans. AB - BACKGROUND: The aims were to investigate cross-culturally the subjective experience of long-term psychiatric patients to determine whether or not they would define their current life situation predominantly in terms of illness. METHOD: The design is a two-by-two comparison by ethnicity (Latino and Euro American) and diagnosis (schizophrenia and unipolar depression) of 80 subjects for DSM-III-R criteria according to the SADS. Patients were interviewed using semi-structured interview (Context of Illness Experience) yielding data coded for qualitative and quantitative analyses. RESULTS: while significant ethnic differences were observed, nearly half the sample did not include illness in their description of their life situation. Other domains (e.g. activities, event) and the extent to which patients perceived themselves as in- or out-of-step with the "rhythm of life" were identified as central patient concerns. CONCLUSIONS: Empirical research on the subjective experiences and representations of the life situation of patients offers clues to the course and treatment of persistent psychiatric disorder. PMID- 9328490 TI - Appraisal, psychological adjustment and expressed emotion in relatives of patients suffering from schizophrenia. AB - BACKGROUND: It is argued that coping theory may be useful in attempting to understand how relatives adopt to the demands of living with a schizophrenia sufferer. METHOD: In a prospective study, univariate and multivariate relationships were explored between appraisal variables (appraisal of symptom threat (primary appraisal) and perceived symptom control (secondary appraisal)) and (a) expressed emotion, and (b) psychological distress in relatives of schizophrenic patients. The profile of relatives who showed sustained distress over time was also examined. RESULTS: The appraisal variables were found to be related to both the concurrent distress (GHQ scores), EE ratings of relatives at the time of the patients' relapse and hospitalization, as well as the subsequent GHQ scores of relatives when the patient was discharged back home. Relatives who showed sustained distress were likely to show high EE and have a longer caring history. CONCLUSIONS: The study gives some support to the theory that appraisal processes underlie how relatives react to having a family member with schizophrenia, and may have implications both for identifying those at risk of poor adaptation, and for understanding strategies that improve well-being. PMID- 9328491 TI - Bereitschaftpotential in schizophrenia. AB - BACKGROUND: Several reports have documented the presence of motor abnormalities in schizophrenic patients. METHOD: Thirty schizophrenics and 28 healthy controls were included in the study. Scalp-recorded bereitschaftpotentials (BPs) generated prior to voluntary movements were recorded in all subjects. RESULTS: The early (NSI) and late components of BP and peak negativity were reduced in all schizophrenic patients. In particular, the NSI was reduced in patients with positive symptoms, and the late component in patients with negative symptoms. CONCLUSIONS: These findings provide further support of the involvement of frontal cortex, subcortical structures and their connections in schizophrenia, and highlight some differences between positive and negative symptom clusters. PMID- 9328492 TI - Intensive in-patient and community intervention versus routine care after attempted suicide. A randomised controlled intervention study. AB - BACKGROUND: A randomised clinical trial was carried out in suicide attempters to assess clinical efficacy of an intensive psychosocial intervention compared with treatment as usual. METHOD: Two hundred and seventy-four suicide attempters presenting for medical treatment were randomly assigned to either intensive psychosocial treatment or 'care as usual'. Intensive psychosocial treatment consisted of brief admission to a special crisis-intervention unit and problem solving aftercare. 'Care as usual' included any form of treatment the assessing clinicians thought appropriate. Psychological well-being was evaluated by the SCL 90 and the Hopelessness Scale at 3, 6 and 12 months following entry in the study. RESULTS: No differences in outcome were found. The probability of repeat suicide attempts in the 12-month follow-up was 0.17 for patients in the experimental group and 0.15 for the control group. There were no differences in ratings on the SCL-90 and the Hopelessness Scale. Patients in the experimental group attended significantly more out-patient treatment sessions. CONCLUSIONS: General implementation of an intensive in-patient and community intervention programme for suicide attempters does not seem justified. PMID- 9328493 TI - One hundred cases of attempted suicide in the elderly. AB - BACKGROUND: Despite the high suicide rate in the elderly, there is a relative lack of information on the longer-term outcome of elderly people who have attempted suicide, particularly their psychiatric and physical morbidity and mortality. METHOD: Comprehensive demographic and psychiatric data were available on 100 consecutive referrals to a liaison psychiatric service of patients over 65 years of age who attempted suicide between 1989 and 1992. Detailed follow-up in 1994 included the interviewing of survivors using GMS-AGECAT. RESULTS: Of the 64 women and 36 men, with a mean age of 75.8 years, 42 subjects were dead at follow up; 12 were suspected suicides, five having died as a delayed result of their index attempt. Twelve women made a further non-lethal suicide attempt. All five male repeat attempts proved fatal. CONCLUSIONS: Elderly people who attempt suicide have a high mortality both from completed suicide and death from other causes. The completed suicide rate is at least 1.5% per year, and the repetition rate is 5.4% per year. Those at risk of further self-harm are likely to be in contact with psychiatric services and to be suffering from persistent depression. PMID- 9328494 TI - Prospective longitudinal study of depression and anosognosia in Alzheimer's disease. AB - BACKGROUND: The aim was to examine the longitudinal evolution of depression and anosognosia in patients with probable Alzheimer's disease (AD). METHOD: Sixty-two of a consecutive series of 116 AD patients that were examined with a structured psychiatric interview had a follow-up evaluation between one and two years after the initial evaluation. RESULTS: At the initial evaluation 19% of the 62 patients had major depression, 34% had dysthymia, and 47% were not depressed. After a mean follow-up of 16 months, 58% of patients with major depression at the initial evaluation were still depressed, whereas only 28% of patients with initial dysthymia and 21% of the non-depressed patients were depressed at follow-up. During the follow-up period, all three groups showed similar declines in cognitive status and activities of daily living. At the initial evaluation, 39% of the patients had anosognosia, and there was a significant increment of anosognosia during the follow-up period. CONCLUSIONS: While dysthymia in AD is a brief emotional disorder, major depression is a longer-lasting mood change. Anosognosia is another prevalent disorder among AD patients, and increases with the progression of the illness. PMID- 9328495 TI - Seasonal changes in psychological well-being in an elderly population. AB - BACKGROUND: Little is known about seasonal fluctuations in psychological well being among elderly people. METHOD: Over a period of 21 months, 1466 elderly people completed the General Health Questionnaire and the Leeds Scales for Depression and Anxiety. Scores during the winter months (December to February) were compared with those during other months of the year. RESULTS: Scores on all scales were significantly higher during the winter months, but there was no difference in rates of caseness. Unlike younger populations, elderly women did not exhibit greater seasonality in well-being than did elderly men. CONCLUSIONS: Elderly people exhibit a small seasonal fluctuation in psychological well-being, which is probably of little clinical importance, and there is no gender difference. The findings support the contention that seasonal mood changes are most pronounced among females of reproductive age. PMID- 9328496 TI - Cost of community care for older people. AB - BACKGROUND: There has been no published study that considers actual costs in a representative sample of people aged > or = 65 years. The present study describes the financial costs of formal community services for elderly people with dementia, depression, anxiety disorder or physical disability. METHOD: Psychiatric morbidity, physical disability and services received were assessed by standardised questionnaire in randomly selected Islington enumeration districts. Subjects were interviewed at home (n = 700). RESULTS: Dementia was the most expensive disorder per sufferer in terms of formal services. Those with depression were also high users of health services. Despite presenting to health services, 90% were not treated with appropriate drugs. In contrast, social services were received by people who were activity-limited or with dementia. The highest service cost for the population as a whole was for the physically disabled. In multivariate analysis the significant predictors of high service costs were living alone, being physically ill, depression, dementia and increasing age. CONCLUSIONS: Failure to detect and treat depression and the anxiety disorders in older people, despite their presentation to medical services, may have major economic consequences as well as contributing to individual suffering. PMID- 9328497 TI - Common mental disorders in primary care in Harare, Zimbabwe: associations and risk factors. AB - BACKGROUND: This study aimed to investigate the associations for common mental disorders (CMD) among primary care attenders in Harare. METHOD: This was an unmatched case-control study of attenders at primary health clinics, general practitioner surgeries and traditional medical practitioner clinics; 199 cases with CMD as identified by an indigenously developed case-finding questionnaire, and 197 controls (non-cases), were interviewed using measures of sociodemographic data, disability, care-giver diagnoses and treatment, explanatory models, life events and alcohol use. RESULTS: CMD was associated with female gender (P = 0.04) and older age (P = 0.02). After adjustment for age, gender and site of recruitment, CMD was significantly associated with chronicity of illness; number of presenting complaints; beliefs in "thinking too much" and witchcraft as a causal model; economic impoverishment; infertility; recent unemployment; an unhappy childhood for females; disability; and consultations with traditional medical practitioners and religious priests. CONCLUSIONS: Mental disorders are associated with female gender, disability, economic deprivation, and indigenous labels of distress states. PMID- 9328498 TI - Postpartum psychiatric illness in Arab culture: prevalence and psychosocial correlates. AB - BACKGROUND: There have been numerous studies of the prevalence of postpartum depression and its putative risk factors in Western Europe and North America, but very few studies in developing countries including the Arab world. METHOD: Ninety five women admitted to the New Dubai Hospital in Dubai, United Arab Emirates, for childbirth were studied. All subjects were assessed in the postpartum period using clinical and socio-cultural instruments: the Self Report Questionnaire (SRQ) at day 2, the Edinburgh Postnatal Depression Scale (EPDS) at day 7, and the Present State Examination (PSE) at week 8 +/- 2 and week 30 +/- 2 after delivery. RESULTS: The prevalence rate of psychiatric morbidity was 24.5% by the SRQ, 17.8% by the EPDS, and 15.8% by the PSE. A number of psychosocial factors emerged as putative risk factors for postpartum depression. CONCLUSIONS: The prevalence rates of postpartum psychiatric morbidity and its risk factors in this Arab culture are similar to the results obtained in numerous previous studies in industrialised countries. These findings have implications for the early detection and care of women at risk for postpartum depression. PMID- 9328500 TI - Efficacy and safety of acamprosate in the treatment of detoxified alcohol dependent patients. A 90-day placebo-controlled dose-finding study. AB - BACKGROUND: Acamprosate is a newly registered drug that appears to reduce alcohol drinking in both animal models and clinical conditions. METHOD: In order to assess the efficacy and safety of the drug in the treatment of detoxified alcoholics, we performed a 90-day double-blind trial comparing two dosages of acamprosate (1332 mg/day and 1998 mg/day). RESULTS: For all efficacy parameters, acamprosate appeared to be significantly superior to placebo, with a trend towards a better effect at the higher dosage. Furthermore, acamprosate appeared to be extremely safe. CONCLUSION: This study confirms that acamprosate could be an interesting adjuvant for maintaining abstinence in detoxified alcoholics. PMID- 9328499 TI - Family planning needs and STD risk behaviours of female psychiatric out-patients. AB - BACKGROUND: There are few studies concerning the family planning needs of female chronic psychiatric patients. We aimed to determine the contraceptive needs and sexually transmitted disease (STD) risk-behaviours of female psychiatric out patients. METHOD: Sixty-six female out-patients with major psychiatric disorders, including schizophrenia, bipolar disorder and mood disorders, completed a semi structured interview (response rate = 63%) and were individually matched for age and ethnicity with 66 women who had never been treated for psychiatric illness. They answered questions on child-rearing and on their methods of contraception in relation to their attitudes towards pregnancy, as well as on their risk for STDs. RESULTS: Compared with controls, the female patients reported having had significantly more induced abortions and were significantly more likely to have given up their own children for others to raise. Heterosexually active psychiatric patients were significantly more likely than controls to have had more than one male sexual partner, to have been pressured into unwanted sexual intercourse, and to report having had sexual intercourse with a suspected bisexual over the preceding year. CONCLUSIONS: These results underscore the priority for developing programmes that reduce female psychiatric patients' risk for unwanted pregnancies and STDs. PMID- 9328501 TI - Randomised controlled trial of psychological debriefing for victims of acute burn trauma. AB - BACKGROUND: Psychological debriefing (PD) is widely used following major traumatic events in an attempt to reduce psychological sequelae. METHOD: One hundred and thirty-three adult burn trauma victims entered the study. After initial questionnaire completion, participants were randomly allocated to an individual/couple PD group or a control group who received no intervention; 110 (83%) were interviewed by an assessor blind to PD status three and 13 months later. RESULTS: Sixteen (26%) of the PD group had PTSD at 13-month follow-up, compared with four (9%) of the control group. The PD group had higher initial questionnaire scores and more severe dimensions of burn trauma than the control group, both of which were associated with a poorer outcome. CONCLUSION: This study seriously questions the wisdom of advocating one-off interventions post trauma, and should stimulate research into more effective initiatives. PMID- 9328502 TI - Eye movement desensitisation and reprocessing versus exposure in vivo. A single session crossover study of spider-phobic children. AB - BACKGROUND: Eye movement desensitisation and reprocessing (EMDR) is a relatively new therapeutic technique that has been proposed as a treatment for post traumatic stress disorder and other anxiety complaints. METHOD: We compared the efficacy of EMDR with that of exposure in vivo in the treatment of a specific phobia. Twenty-two spider-phobic children who met the DSM-III-R criteria for specific phobia participated in the study. Children were treated with one session of exposure in vivo and one session of EMDR in a crossover design. Treatment outcome was evaluated by self-report measures, a behavioural avoidance test and a physiological index (skin conductance level). RESULTS: Results showed positive effects of EMDR, but also suggest that it is especially self-report measures that are sensitive to EMDR. Improvement on a behavioural measure was less pronounced, and exposure in vivo was found to be superior in reducing avoidance behaviour. With regard to skin conductance level, EMDR and exposure in vivo did not differ. CONCLUSIONS: EMDR has no additional value in treatment of this type of animal phobia, for which exposure in vivo is the treatment of choice. PMID- 9328503 TI - Discontinuation rates of SSRI's and tricyclic antidepressants. PMID- 9328504 TI - Discontinuation rates of SSRI's and tricyclic antidepressants. PMID- 9328505 TI - Cost effectiveness of antidepressant treatment. PMID- 9328506 TI - Cognitive function and fall-related fractures. PMID- 9328507 TI - Clozapine-induced hypersalivation. PMID- 9328508 TI - Clozapine, Chinese and blood. PMID- 9328509 TI - Citalopram-induced decreased libido. PMID- 9328510 TI - Clozapine treatment, eosinophilia and agranulocytosis. PMID- 9328511 TI - Clozapine monotherapy and ketoacidosis. PMID- 9328512 TI - Sigmund: a European database of mental health surveys. PMID- 9328513 TI - Influenza and schizophrenia. PMID- 9328514 TI - Assessing risk in the mentally disordered. Introduction. PMID- 9328515 TI - Risk assessment and clinical risk management: the lessons from recent inquiries. PMID- 9328516 TI - The epidemiology of crime, violence and schizophrenia. PMID- 9328517 TI - The investigation of acting on delusions as a tool for risk assessment in the mentally disordered. PMID- 9328518 TI - Predictors of risk in serious sex offenders. PMID- 9328519 TI - Patients as parents: the risk to children. PMID- 9328520 TI - Assessing dangerousness: protecting the interests of patients. PMID- 9328521 TI - Assessing risk: are we being overcautious? PMID- 9328522 TI - Practical aspects of clinical risk assessment and management. PMID- 9328523 TI - Training trainers in risk assessment. PMID- 9328524 TI - Risk assessment in a climate of litigation. PMID- 9328525 TI - Determination of pyrrolizidine alkaloids in honey from selected sites by solid phase extraction and HPLC-MS. AB - A method was developed for the determination in honey of the Ragwort (Senecio jacobaea) derived pyrrolizidine alkaloids jacoline, jacozine, jacobine, seneciphylline and senecionine, combining solid-phase extraction with high performance liquid chromatography and atmospheric pressure chemical ionization mass spectrometric detection. The method allowed determination of individual alkaloids and offered a considerable improvement in terms of speed, sensitivity and specificity over previous approaches, but was not suitable for determination of jaconine, a minor alkaloid in Ragwort. Instrument calibrations were linear over the range 0.005 to 100 micrograms/ml, equivalent to approximately 0.001 to 2.0 mg/kg in honey with the extraction method used and allowing for observed recoveries. Detection limits in honey were 0.002 mg/kg. Recoveries for most of the alkaloids were between 57 and 70%. The alkaloids have been determined in a number of samples of honey selected after pollen identification and counting. The alkaloids were not detectable in samples containing two grains or less of Ragwort pollen per gram of honey. Samples collected in late July and August contained Ragwort pollen at 15-21 grains/g and total alkaloid concentrations of 0.011-0.056 mg/kg. Similar contributions to the total were made by jacozine, seneciophylline and senecionine, with jacobine making a larger and jacoline a smaller contribution. Two samples of honey containing Ragwort pollen at 24 and 16 grains/g had total alkaloid concentrations of 0.42 and 1.48 mg/kg respectively (not corrected for recovery). The alkaloid profile in these samples was dominated by seneciphylline and senecionine which together comprised 90-95% of the total. Alkaloids were not detected in retail honeys. PMID- 9328526 TI - The influence of deck storage and initial processing on patulin levels in apple juice. AB - Patulin, a secondary metabolite produced by Penicillium expansum and some other fungal species, is a common contaminant of ripened apples used for the production of apple juice concentrates. The limited availability of suitable storage facilities may result in fruit being subjected to storage in the open ('deck storage') for extended periods of time, prior to processing. A study was conducted to determine the influence that deck storage and subsequent initial processing practices had on patulin levels in freshly pressed juice. Over the study period, triplicate samples were collected at four strategic processing points from individual consignments of Granny Smith apples deck-stored for 7, 15 and 33 days, respectively. Over the study period, mean patulin levels in non processed fruit increased from 90 to 2445 ng/g, respectively, but decreased to between 75 and 695 ng/g, respectively, following a water wash step. Subsequent removal of rotten/damaged fruit decreased patulin levels further (to between 55 and 405 ng/g, respectively), although the numerical decreases between sampling points were not shown to be statistically significant (P > 0.05). However, patulin levels were significantly higher (P < 0.05) in the rejected rotten/damaged fruit (mean levels ranged from 1120 to 6235 ng/g, respectively). P. expansum was the major patulin-producing fungus isolated from the juice samples. The mycological analyses tended to support the chemical data, in that removal of the rotten/damaged fractions significantly reduced total fungal counts in the juice samples. PMID- 9328527 TI - Determination of the fate of three Fusarium mycotoxins through wet-milling of maize using an improved HPLC analytical technique. AB - The fate of three Fusarium mycotoxins, nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN), all common contaminants in New Zealand-grown maize, has been measured in fractions of maize after passage through a commercial wet-milling plant. Distribution of the three toxins follows a pattern reasonably expected from their physical solubility characteristics. The highly water-soluble mycotoxins, NIV and DON, were found at high concentrations (up to 8.8 mg/kg) in concentrated steep liquor (CSL) fractions, but at low levels (less than 0.3 mg/kg) in the solid (germ, fibre and gluten) fractions. The converse was true for ZEN, which is relatively insoluble in water. For ZEN, the maximum concentration found in CSL was 0.6 mg/kg compared with 2.2-4.8 mg/kg in germ, fibre and gluten fractions. Accordingly, an animal food byproduct composed mainly of pressed fibre and concentrated steep liquor was usually found to contain concentrations of all three mycotoxins above those existing in the input maize. A single sample of corn oil recovered during the study also had a high concentration (4.6 mg/kg) of ZEN. The analytical clean-up method used converts all trichothecenes present to parent alcohols, therefore results are indicative of total trichothecene content. HPLC analytical conditions suitable for the analysis of NIV and DON in complex process grain products are also described. PMID- 9328528 TI - Evaluation of enzyme-linked immunosorbent assay for analysis of beer for fumonisins. AB - A recently developed sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) was applied to the determination of fumonisins in beer. Intra-assay and inter-assay recoveries averaged 98.7-102.8% at added fumonisin B1 (FB1) levels of 0.5-50 ng/ml beer, and coefficients of variation were 2.8-4.4 and 4.7 8.6%, respectively. Cross-reactivity of fumonisin B2 (FB2) compared with FB1 averaged 67% in beer. Two experiments were carried out to compare ELISA with liquid chromatography (LC) for determination of fumonisins in beer. In the first experiment, 19 samples (five previously known positive, nine other samples and five spiked samples) were passed through commercial immunoaffinity columns (ICs) and analysed by LC before conducting blind ELISA determinations on the extracts and beers directly. The known positive beers and extracts were used as blind duplicates. The second comparative experiment screened 46 beer samples by ELISA and then 22 positive and three of the negative samples were analysed by LC; the highest level found was 64.3 ng total fumonisins/ml measured by LC (24.7 ng/ml by ELISA). Regression analyses showed good correlation between ELISA and LC in the first experiment but low level interferences (equivalent to up to 5.35 ng fumonisin/ml) were observed by ELISA in the IC extracts. Five of nine beers negative by LC showed < 1 ng/ml ELISA responses on direct beer analysis. The second comparative experiment indicated underestimation by ELISA. However, there were two samples which tested positive by ELISA (0.2 ng/ml) but were found negative by LC (results close to the detection limits of both methods, which were 0.1 or 0.2 ng/ml by ELISA and 0.1-0.15 ng each fumonisin/ml by LC). There were no false negatives. It is concluded that ELISA has considerable value in rapid screening of beer directly for fumonisins. PMID- 9328529 TI - Occurrence of aflatoxin M1 in commercial pasteurized milk determined with ELISA and HPLC. AB - Eighty-one samples of commercial pasteurized milk from Athens market were analysed for the presence of aflatoxin M1 (AFM1). A combination of a commercial ELISA kit and a modified HPLC method was applied for the rapid and reliable determination of AFM1. AFM1 concentrations in milk extracts were initially estimated by ELISA. Samples found to contain more than 5 ng/l were further quantitated by HPLC. Determination was performed after derivatization of AFM1 to its hydroxylated product AFM2a. The recovery of the HPLC method used was found to be close to 100%. Thirty-two samples contained aflatoxin M1 at levels of 2.5-5 ng/l, none contained more than 5 ng/l, while 31 contained only traces of aflatoxin (0.5-1 ng/l). In nine samples no AFM1 was detected. There was no seasonal influence on the aflatoxin content of the milk samples analysed. PMID- 9328530 TI - Direct synthesis of aflatoxin B1-N7 guanine adduct: a reference standard for biological monitoring of dietary aflatoxin exposure in molecular epidemiological studies. AB - Aflatoxin B1-N7-guanine and aflatoxin B1-human serum albumin adducts have been established as biomarkers of dietary aflatoxin exposure in epidemiological studies. Earlier chemical oxidants were used to synthesize aflatoxin B1-8,9 epoxide in vitro and its subsequent interaction with DNA or synthetic oligodeoxynucleotide was used as a source of authentic aflatoxin B1-N7-guanine adduct. In the present communication we report a simple single step procedure for the synthesis of aflatoxin B1-N7-guanine adduct using free guanine and m chloroperbenzoic acid as the chemical oxidant for the production of AFB1-8,9 epoxide. At a molar ratio of 1:1 of AFB1-8,9-epoxide and guanine the recovery of the AFB1-N7-guanine adduct was found to be 60% while at higher molar ratios (1:2 and 1:4) of guanine the recovery of the AFB1-N7-guanine adduct was found to be low (30-40%). HPLC analysis of the AFB1-N7 guanine adduct showed a retention time identical with the retention time of the AFB1-N7-guanine adduct synthesized using calf thymus DNA. TLC-fluorodensitometric analysis indicated that the Rf of the AFB1-N7-guanine adduct was zero. Spectral analysis of the adduct synthesized showed an excitation wavelength of 360 nm and emission wavelength at 440 nm in phosphate buffer (100 mM, pH 7.4). Further, the formation of the AFB1-N7-guanine adduct was confirmed by perchloric acid treatment resulting in the destruction of the adduct. The AFB1-N7-guanine adduct thus synthesized was stable in both acidic as well as lyophilized conditions over a period of 2 weeks. The antibody capture assay showed that the antibodies produced against the antigen BSA-guanine-N7-AFB1 also cross-reacted with calf thymus DNA-AFB1 adduct, indicating specificity to the guanine-N7-AFB1 moiety. The method developed may find immediate application as a source of authentic reference standard in molecular epidemiological studies. PMID- 9328532 TI - Cadmium variations in Manchego cheese during traditional cheese-making and ripening processes. AB - Variations in cadmium content were determined throughout cheese manufacturing and ripening processes by applying graphite furnace atomic absorption spectrophotometry to samples of natural pasteurized milk, rennet, curd whey, pressed curd, pressing whey and cheese. The total mean cadmium contents were 4.79 +/- 2.4 and 4.67 +/- 2.1 microgram/kg fresh weight for newly-made and mature cheeses respectively. ANOVA revealed statistically significant differences (p < 0.001) in cadmium levels (fresh weight) and these differences were due to the influence of moisture content during cheese manufacture, since no statistically significant differences (p > 0.05) were found for dry weight. Nevertheless, cadmium levels based on dry weight increased during pasteurization and more noticeably on ferment addition. ANOVA performed during the ripening process revealed significant differences between portions and ripening times for both fresh and dry weights. By Tukey's test (p < 0.05) for portions, two homogeneous groups were established, one corresponding to the outer portion with a greater cadmium content and the other comprising the middle and inner portions. The contribution of cadmium to Spanish mean intake is between 0.098 and 0.147 micrograms/week for new cheese and between 0.168 and 0.245 micrograms/week for mature cheese. PMID- 9328531 TI - Comparison of supercritical fluid extraction and conventional liquid-solid extraction for the determination of benzo[a]pyrene in water-soluble smoke. AB - Extraction of benzo[a]pyrene from 12 samples of water-soluble liquid smokes by means of a conventional liquid-solid extraction method was compared with extraction with a supercritical fluid. Results were satisfactory for both methods, there being no significant differences between the recoveries and precisions obtained for each one. The merits and disadvantages of each extraction method are discussed. PMID- 9328533 TI - Preliminary assessment of potential health hazards associated with barium leached from glazed ceramicware. AB - Ceramic glazes contain several elements which have the potential to leach into food or beverages that are held or stored in ceramicware. Recently, barium salts have been investigated as one of the alternatives to lead in frit formulations for glazes. This preliminary evaluation addresses the potential health hazards associated with barium at levels that might leach from glazed ceramicware. A set of specialty ceramicware, consisting of five teacups and a pitcher, was examined for extractable barium. Exposure to barium that adults (18-44 years) might encounter using the vessels for coffee, tea, or orange juice was estimated. The exposure estimate was derived from values for intakes of the beverages and for the barium migration from glazed ceramicware test samples. An established reference dose (RfD) for barium exposure for the critical effect of hypertension was identified. The potential hazard associated with the leaching of barium from glazed ceramicware varied with the level of use. Consuming beverages in amounts up to the 95th percentile would not result in total barium intake in amounts that exceed the RfD; consuming large quantities (> 95th percentile) of beverages such as tea or coffee from glazed vessels might. This suggests that for a small portion of the population of users, intake of barium may be in quantities that warrant further consideration as a potential health hazard. Analyses of a broad sample of ceramicware and study of barium leaching behaviour under actual use conditions are needed to assess further the significance of these findings. PMID- 9328535 TI - Prediction of worst case migration from packaging to food using mathematical models. AB - Prediction of migration from packaging to food is often made using equations which are not always designed specifically for the problem. At least, these equations should overestimate migration, in order to be on the safe side. Integration of Fick's equation under the assumption of 'infinite packaging' provides an equation which is very practical since it requires only a few experimental data. It is shown here that, unfortunately, the use of this equation leads to a systematic underestimation of the diffusivity, by the square of the percentage of migration at steady state. In contrast to widely accepted opinion, this model is not conservative. A conservative approach requires that the diffusivity is determined under 'finite packaging' assumptions, associated with very large volumes of food and with long term experiments. These equations are applied to the migration of a phenolic antioxidant from polypropylene. PMID- 9328534 TI - Migration of residual contaminants from secondary recycled poly(ethylene terephthalate) into food-simulating solvents, aqueous ethanol and heptane. AB - This study measured the migration of benzene, butyric acid, dodecane, octadecane, tetracosane, diazinon, lindane, and copper (II) ethyl hexonate from poly(ethylene terephthalate)(PETE) sheets into the food simulants, 8% ethanol/water and n heptane. The contaminated PETE sheets were extruded from PETE chips that had been previously contaminated but were washed, dried, and remelted. The level of these contaminants remaining in the extruded sheets ranged from benzene at 0.6 mg/kg to copper salt at 24 mg/kg. The extraction data demonstrate that migration of the residual contaminants from the extruded PETE sheets resulted in concentrations lower than 10 micrograms/kg in the food simulants. At very high residual concentrations of butryic acid (147 mg/kg) and benzene (218 mg/kg) in sheets made from unwashed PETE, higher amounts of the contaminant migrated into the food simulants. This migration resulted in contaminant concentrations exceeding 10 micrograms/kg and suggests that unwashed recycled PETE may not comply with FDA requirements. The crystallinity of extruded PETE sheets in this study ranged from 5 to 15%, which is lower than that of most commercial PETE (30%). Therefore, the migration data obtained from these test samples represent the most severe conditions for conservative exposure evaluations. PMID- 9328536 TI - The bioavailability of residues of the furazolidone metabolite 3-amino-2 oxazolidinone in porcine tissues and the effect of cooking upon residue concentrations. AB - Residues of furazolidone in pig tissues have previously been shown to be bioavailable in the rat. However, no specific furazolidone metabolite has been identified in the tissues of a second species. Tissues were taken from pigs that had been treated therapeutically with furazolidone, lyophilized and then fed to female Sprague Dawley rats for 3 days. Protein-bound and solvent-extractable residues containing the side chain metabolite 3-amino-2-oxazolidinone (AOZ) were detected in the liver, kidney and muscle of the rats using HPLC-thermospray mass spectrometry. Furazolidone-contaminated pig tissues which had undergone solvent extraction and thereby contained only bound residues, was fed to two rats. Bound and extractable AOZ was detected in liver, kidney and muscle. Since it is most likely that consumers would eat animal tissue which had been cooked, an experiment was carried out to determine the effects of cooking upon the concentrations of AOZ residues in pig tissues. Total AOZ concentrations were not significantly reduced in liver, kidney or muscle, following frying, grilling or microwaving. PMID- 9328537 TI - Seasonal variation in the prevalence of Down syndrome at birth: a review. AB - STUDY OBJECTIVE: Many studies on seasonality in Down syndrome (DS) have been performed and have come to different conclusions. It is suggested that seasonal variation in hormone production by the hypothalamus-pituitary-ovarian axis just before ovulation leads to seasonality in conception rates of DS. This study aimed to determine whether there is seasonal variation in the prevalence of DS at birth as a proxy for seasonality in DS at conception. DESIGN: All the English and Dutch articles on this topic were reviewed. Articles published between 1966 and January 1996 were traced by Medline, and by the reference lists. MAIN RESULTS: Twenty articles met the criteria for inclusion. Although seven of these studies reported seasonality in DS prevalence, no consistent seasonal pattern was found in DS at birth in these studies, or in the remaining studies. A seasonal pattern could not have been masked by the effects of maternal age, induced abortions, shortened gestation, or misclassification of DS. CONCLUSION: Seasonality in the prevalence of DS at birth does not exist. Evidence did not support the suggestion that DS occurrence is related to seasonality in hormone production. PMID- 9328538 TI - Putting trials on trial--the costs and consequences of small trials in depression: a systematic review of methodology. AB - STUDY OBJECTIVE: To determine why, despite 122 randomised controlled trials, there is no consensus about whether the selective serotonin reuptake inhibitors or tricyclic and related antidepressants should be used as first line treatment of depression. DESIGN: Systematic review of all RCTs comparing selective serotonin reuptake inhibitors and tricyclic or heterocyclic antidepressants. MAIN RESULTS: The shortcomings identified in the 122 trials were as follows: (1) there was inadequate description of randomisation, (2) the outcomes used were mainly observer rated measurements of depression, and studies failed to use quality of life measures or perform economic evaluations, (3) doses of tricyclic antidepressants were inadequate, (4) generalisability of studies was poor (including a reliance on secondary care settings and inadequate follow up), and (5) there were statistical shortcomings such as low statistical power, failure to use intention to treat analyses, and the tendency to make multiple comparisons. CONCLUSIONS: Future RCTs should be designed to inform policy makers and address these methodological shortcomings. PMID- 9328539 TI - How should interventions to reduce inequalities in health be evaluated? AB - OBJECTIVE: The effectiveness of interventions which have been proposed or are currently in progress to reduce socioeconomic inequalities in health is largely unknown. This paper aims to develop guidelines for evaluating these interventions. APPROACH: Starting from a set of general guidelines which was recently proposed by a group of experts reporting to the national Programme Committee on Socioeconomic Inequalities in Health in The Netherlands, an analysis was made of the appropriateness of different study designs which could be used to assess the effectiveness of interventions to reduce inequalities in health. RESULTS: A "full" study design requires the measurement, in one or more experimental populations and one or more control populations, of changes over time in the magnitude of socioeconomic inequalities in health. This will usually imply a community intervention trial. Five alternative study designs are distinguished which require less complex measurements but also require more assumptions to be made. Several examples are given. CONCLUSIONS: Building up a systematic knowledge base on the effectiveness of interventions to reduce socioeconomic inequalities in health will be a major enterprise. Elements of a strategy to increase learning speed are discussed. Although the guidelines and design recommendations developed in this paper apply to the evaluation of specific interventions where rigorous evaluation methods can often be used, they may also be useful for the interpretation of the results of less rigorous evaluation studies, for example of broader policies to reduce socioeconomic inequalities in health. PMID- 9328540 TI - Validation in London of a physical activity questionnaire for use in a study of postmenopausal osteopaenia. AB - STUDY OBJECTIVE: To determine the validity of a self administered physical activity questionnaire to be used as part of a screening device for postmenopausal osteopaenia (with additional questions on medical history and calcium intake). DESIGN: A questionnaire was posted to 86 perimenopausal women to enquire about weekly hours spent in non-sedentary activity at work, in the household, and during leisure hours. Subjects who returned the questionnaire were visited at home and asked to complete a four day activity diary and subsequently to undertake a submaximal estimate of VO2 MAX, carried out using a treadmill ergometer. They were interviewed to clarify questionnaire and diary entries. Questionnaire validity was assessed in comparison with the diary estimates of hours of activity and with VO2 MAX. PARTICIPANTS: A total of 86 perimenopausal women aged 43-54 years were randomly selected from a GP list in Hammersmith, London. Thirty five women (41%) returned the questionnaire. They were visited at home, given the diary to complete, and invited to attend the physiology laboratory for VO2 MAX measurements. Twenty six of the 35 (74%) completed the study and were included in the final analysis. MAIN RESULTS: Women spent an average of 51 hours per week in non-sedentary activities. Questionnaire and diary yielded similar results (51.05 versus 51.30 h/wk), and there was a good correlation between diary and questionnaire estimates of total weekly hours of non-sedentary activity (r = 0.45, p < 0.05). Other significant correlations were for standing (r = 0.69, p < 0.01), leisure activities (r = 0.66, p < 0.01), and for light household activities (r = 0.42, p < 0.05). Correlations were better for employed than non-employed subjects. In relation to the diary, the questionnaire correctly classified 60% into the top or bottom half of the distribution activity. Sensitivity and specificity of the questionnaire were both equal to 61.5%. CONCLUSIONS: The questionnaire is useful for classifying subjects according to their level of activity, especially when administered in conjunction with an interview. The four day diary provided a useful reference measure and a focus for discussing activity patterns during an interview related to the questionnaire responses. PMID- 9328541 TI - Why do women doctors in the UK take hormone replacement therapy? AB - STUDY OBJECTIVES: To ascertain the determinants and experiences of hormone replacement therapy (HRT) use by menopausal women doctors. DESIGN: Postal questionnaire. SETTING: UK. PATIENTS: A randomized stratified sample of women doctors who obtained full registration between 1952 and 1976, taken from the current Principal List of the UK Medical Register. MAIN OUTCOME MEASURES: Current and previous use of HRT; reasons for and against HRT use; menopausal status; hormonal contraceptive use; lifestyle patterns; family and personal history of disease. MAIN RESULTS: While 73.2% of 471 users had started HRT for symptom relief, 60.9% cited prevention of osteoporosis and 32.7 prevention of cardiovascular disease. Altogether 18.7% had started for preventive purposes alone. Significant predisposing factors to starting HRT were the presence and severity of menopausal symptoms, surgical menopause, past use of hormonal contraception, and a family history of osteoporosis. HRT users were also more likely to use skimmed rather than full fat milk, to try to increase their intake of fruit, vegetables, and fibre, and to undertake vigorous physical activity at least once a week. They were less likely to have had breast cancer. Long duration users were more likely than short duration users to be past users of hormonal contraception and to be using HRT for prevention of osteoporosis as well as symptom relief; they were less likely to have experienced side effects. CONCLUSIONS: The high usage of HRT by women doctors reflects the fact that many started HRT on their own initiative and with long term prevention in mind. The results may become generalisable to the wider population as information on the potential benefits of HRT is disseminated and understood. However, HRT users may differ slightly from non-users in health-related behaviour and a substantial minority may never take up HRT, at least until the benefit-risk ratio is more clearly established. PMID- 9328543 TI - The changing relationship between prescribing and unemployment at family health service authority level in England, 1983-92. AB - STUDY OBJECTIVE: To investigate the relationship between unemployment and prescribing costs over time. DESIGN: This was a longitudinal study. SETTING: All 90 family health service authorities in England, 1983-92. PARTICIPANTS: All general practices in England. MAIN RESULTS: The strength of the relationship varied over the period, falling to a very low value during the last two years of the decade. CONCLUSION: Unemployment rates are not suitable as a proxy for the determination of prescribing costs. PMID- 9328542 TI - Enthusiasm or uncertainty: small area variations in the use of mammography services in Ontario, Canada. AB - STUDY OBJECTIVE: To examine the variation in mammography utilisation in relation to age group and indication across health planning regions in Ontario, Canada. DESIGN: This study includes all women aged 30 and over in Ontario who received a mammogram between July 1, 1990 and December 31, 1991. Data from a sample of 10,000 women aged 50-69 were used to assign mammogram indication as "screening", "possible diagnostic", or "probable diagnostic" based on previous health care utilisation patterns. Age specific rates and age adjusted rates in relation to age group (30-39, 40-49, 50-69, and 70 + years) and region were derived. MAIN RESULTS: Overall, 572,762 women received one or more mammograms. Rates increased from 30-54 years and decreased thereafter. Similar variations were seen in the 40 49 and 50-69 age groups. The ranking in the area specific rates remained consistent for all ages except the 30-39 year range. In relation to indication, the largest variation across regions occurred in the screening group. CONCLUSIONS: Mammography utilisation varies across age groups. The greatest variability is for screening, probably because of physician referral patterns, patient uptake, and perhaps access to mammography. The extent of variation was similar when compared between groups where recommendations were consistent (ages 50-59) and where they were inconsistent (ages 40-49) suggesting that perhaps enthusiasm rather than uncertainty is related to regional variation for this procedure. PMID- 9328544 TI - A confidential enquiry into emergency hospital admissions on the Isle of Wight, UK. AB - OBJECTIVES: To quantify the proportion of potentially avoidable emergency short term admissions to hospital and to identify ways in which they could have been avoided. DESIGN: Confidential enquiry by peer review group. SETTING: St Mary's Hospital, Newport, Isle of Wight. SUBJECTS: All emergency, short term admissions (discharged home within five days) to medicine, general surgery, orthopaedics, gynaecology, ENT, and ophthalmology specialties for 28 (24 hour) days over a six month period in 1994. MAIN OUTCOME MEASURES: Appropriateness of admissions decided by the peer group, the peer group's opinion of ideal management, and the patients' views on the appropriateness of their admission. RESULTS: Altogether 139 cases satisfied the inclusion criteria. Complete data were collected on 123 cases and the peer group considered 81 in the time available. Twenty one of the 81 cases were judged "potentially avoidable". These represent 9.5% (95% CI 6.3%, 13.5%) of short term admissions to the specialties studied. The peer group considered that seven of 10 patients referred by a general practitioner (GP) could have been managed by the GP alone and that the remaining three had been referred appropriately but need not have been admitted had a consultant opinion been available in the accident and emergency (A&E) department. Two of the 10 would have required home support to avoid hospital admission. Five of 11 patients who referred themselves to A&E could have been discharged home without admission and without recourse to a specialist opinion. The remaining six could have been discharged had a consultant opinion been available in A&E. CONCLUSIONS: Urgent consultant opinion, either in A&E or in an outpatient clinic, would have prevented most of these inappropriate admissions, and home support would have expedited the ability to discharge some patients. Further research into the costs and benefits of methods for providing these services is needed urgently. PMID- 9328545 TI - Iodine, milk, and the elimination of endemic goitre in Britain: the story of an accidental public health triumph. AB - OBJECTIVE: To determine how iodine deficiency and endemic goitre disappeared in Britain. DESIGN: Review of surveys of endemic goitre and iodine nutrition. MAIN RESULTS: Endemic goitre was widespread in Britain but has declined, most notably since the 1960's. Its disappearance was probably due to changes in farming practice, especially iodine supplementation in dairy herds which has resulted in iodine contamination of milk and dairy produce. CONCLUSIONS: Although iodization of dairy herds offers an indirect method of counteracting iodine deficiency, it is haphazard and there should be careful and continuous monitoring of iodine intakes in the population. PMID- 9328546 TI - Central obesity, insulin resistance, syndrome X, lipoprotein(a), and cardiovascular risk in Indians, Malays, and Chinese in Singapore. AB - STUDY OBJECTIVE: To examine the hypothesis that the higher rates of coronary heart disease (CHD) in Indians (South Asians) compared with Malays and Chinese is at least partly explained by central obesity, insulin resistance, and syndrome X (including possible components). DESIGN: Cross sectional study of the general population. SETTING: Singapore. PARTICIPANTS: Random sample of 961 men and women (Indians, Malays, and Chinese) aged 30 to 69 years. MAIN RESULTS: Fasting serum insulin concentration was correlated directly and strongly with body mass index (BMI), waist-hip ratio (WHR), and abdominal diameter. The fasting insulin concentration was correlated inversely with HDL cholesterol and directly with the fasting triglyceride concentration, blood pressures, plasminogen activator inhibitor 1 (PAI-1), and tissue plasminogen activator (tPA), but it was not correlated with LDL cholesterol, apolipoproteins B and A1, lipoprotein(a), (Lp(a)), fibrinogen, factor VIIc, or prothrombin fragment (F)1 + 2. This indicates that the former but not the latter are part of syndrome X. While Malays had the highest BMI, Indians had a higher WHR (men 0.93 and women 0.84) than Malays (men 0.91 and women 0.82) and Chinese (men 0.91 and women 0.82). In addition, Indians had higher fasting insulin values and more glucose intolerance than Malays and Chinese. Indians had lower HDL cholesterol, and higher PAI-1, tPA, and Lp(a), but not higher LDL cholesterol, fasting triglyceride, blood pressures, fibrinogen, factor VIIc, or prothrombin F1 + 2. CONCLUSIONS: Indians are more prone than Malays or Chinese to central obesity with insulin resistance and glucose intolerance and there are no apparent environmental reasons for this in Singapore. As a consequence, Indians develop some but not all of the features of syndrome X. They also have higher Lp(a) values. All this puts Indians at increased risk of atherosclerosis and thrombosis and must be at least part of the explanation for their higher rates of CHD. PMID- 9328547 TI - Lipid profile and socioeconomic status in healthy middle aged women in Sweden. AB - STUDY OBJECTIVE: To examine the relationship between socioeconomic status (SES) and full lipid profile in middle aged healthy women. PARTICIPANTS: These comprised 300 healthy Swedish women between 30 and 65 years who constitute the control group of the Stockholm female coronary risk study, a population based, case-control study of women with coronary heart disease (CHD). The age matched control group, drawn from the census register of greater Stockholm, was representative of healthy Swedish women aged 30-65 years. Five measures of SES were used; educational level, occupation, decision latitude at work, annual income, and size of house or apartment. MAIN RESULTS: Swedish women with low decision latitude at work, low income, low educational level, blue collar jobs, and who were living in small houses or apartments had an unhealthy lipid profile, suggesting an increased risk of CHD. Part of this social gradient in lipids was explained by an unhealthy lifestyle, but the lipid gradients associated with decision latitude at work and annual income were independent of these factors. Decision latitude, educational level, and annual income had the strongest associations with lipid profile. These associations were independent of age, menopausal status, smoking, sedentary lifestyle, alcohol consumption, obesity, excess abdominal fat, and unhealthy dietary habits. Of the lipid variables, low high density lipoprotein cholesterol (HDL) levels were most consistently associated with low SES. CONCLUSIONS: Decision latitude at work was the strongest SES predictor of HDL levels in healthy middle aged Swedish women, after simultaneous adjustment for other SES measures, age, and all lifestyle factors in the multivariable regression model. PMID- 9328548 TI - Asthma mortality in Australia 1920-94: age, period, and cohort effects. AB - STUDY OBJECTIVE: To investigate asthma mortality during 1920-94 in Australia in order to assess the relative role of period and birth cohort effects. DESIGN: Asthma mortality (both sexes) was age standardised and examined for changes over time. The data were also examined for age, period, and cohort (APC) effects using Poisson regression modelling. SETTING: National Australian mortality data. PARTICIPANTS: Population (both sexes) aged 15-34 years, 1920-94. MAIN RESULTS: Age adjusted period rates indicate an increase in asthma mortality during the 1950's, and increases and subsequent falls (epidemics) during the mid 1960s and late 1980s. APC modelling suggested an increasing cohort effect (adjusted for both age and period) from the birth cohort 1950-54 onwards. Period effects (adjusted for age and cohort) are characterized by an increase in the 1950s (possibly due to changes in diagnostic labelling), minimal or no increases in the mid 1960s and late 1980s (where period peaks had been noted when data were adjusted for age only), and declines in mortality risk subsequent to the periods where age-period analysis had noted increases. Thus, in Australia, some of the mid 1960s epidemic in asthma deaths, and all of the late 1980s mortality increase, seem to be attributable to cohort effects. CONCLUSIONS: The increase in asthma mortality cohort effect is consistent with empirical evidence of recent increases in prevalence (and presumably incidence) of asthma in Australia, and suggests the need for more research into the underlying environmental aetiology of this condition. PMID- 9328549 TI - Time trend and age-period-cohort effects on gastric cancer incidence in Zaragoza and Navarre, Spain. AB - STUDY OBJECTIVE: To describe time trends in gastric cancer incidence in Zaragoza and Navarre, and to investigate time period and birth cohort as determinants of such trends. DESIGN: Cases from two registries were grouped into five year intervals and the following were calculated: age specific and sex specific incidence rates, and the male to female ratio. Log linear models including age, period of diagnosis, and birth cohort were fitted. SETTING: The Zaragoza Cancer Registry covers the province of Zaragoza, which has a population of 824,776 (403,755 men and 421,021 women). The Navarre Cancer Registry covers the province of Navarre which has 512,512 inhabitants (254,786 men and 257,726 women). In both cases population figures were based on the late census. PATIENTS: These comprised incident cases of gastric cancer reported to the Zaragoza Cancer Registry in 1963 87 and to the Navarre Cancer Registry in 1973-87. MAIN RESULTS: Navarre registered higher adjusted and cumulative rates than Zaragoza for both sexes. In both provinces, there were relative declines in the rates for men and women of 3% and 4% respectively per year. In Zaragoza, the risk of developing stomach cancer fell in generations born between 1888 and 1933, and rose in subsequent birth cohorts in both sexes, while in Navarre the cohort effect showed an approximately linear risk for both sexes. Both provinces recorded increases in risk associated with cohorts born between 1933 and 1943. CONCLUSION: The incidence rates of gastric cancer fell in both Zaragoza and Navarre. The reason for the greater incidence of gastric cancer in Navarre remains unknown. Trends in rates seem to be mainly linked to birth cohort. Increases in risk in generations born after 1933 may be ascribable to nutritional deficiencies in the early years of life. PMID- 9328550 TI - Cancer survival in Estonian migrants to Sweden. AB - OBJECTIVE: To quantify the eventual extra loss of life incurred to cancer patients in Estonia compared with those in Sweden that was possibly attributable to differences in society. DESIGN: Population based survival of cancer patients in Estonia was compared with that of Estonian immigrants to Sweden and that of all cancer patients in Sweden. The cancer sites studied were female breast and ovary, male lung and prostate, and male and female stomach and colon. SETTING: Data on incident cases of cancer were obtained from the population based Swedish and Estonian cancer registries. PARTICIPANTS: Data from Estonian patients in Sweden, Estonian patients in Estonia, and patients from the total Swedish population were included in the study. MAIN RESULTS: Differences in survival among the three populations, controlling for follow-up time and age at diagnosis, were observed in breast, colon, lung, ovarian, and prostate cancers. The survival rates of Estonians living in Sweden and the total population of Sweden were better than that of the Estonians living in Estonia. For cancers of the breast and prostate, the excess mortality in the older age group (75 and above) was much greater in Estonia than in the other populations. CONCLUSIONS: Most differences in cancer survival between Estonian and Swedish populations studied could be attributed to a longer delay in diagnosis, and also to inferior treatment (including access to treatment) in Estonia compared with Sweden. Estonia's lag in socioeconomic development, particularly in its public health organisation and funding, is probably the main source of the differences observed. PMID- 9328551 TI - Secular trends in proximal femoral fracture, Oxford record linkage study area and England 1968-86. AB - OBJECTIVE: To study hospital admission rates for fractures of the proximal femur over a period when incidence is reported to have increased, compensating for known lack of precision in coding, excluding nonemergency admissions and transfers, and modelling for age, period, and cohort effects. DESIGN: Validation of coding of a sample of hospital admissions followed by study of two sets of routinely collected statistical abstracts of hospital records; graphical analysis and statistical modelling were used to search for period and cohort effects. SETTING: Oxfordshire and west Berkshire in 1968-86, covered by the Oxford record linkage study (ORLS), and ENGLAND in 1968-85, covered by the hospital inpatient enquiry (HIPE). The ORLS and HIPE datasets are almost independent (ORLS contributed about 1.8% of the HIPE data). SUBJECTS: Records of patients aged 65 and over. OUTCOME MEASURES: Admission rates for fractured neck of femur and fracture of other and unspecified parts of femur (N820 and N821), and evidence of period and cohort effects. RESULTS: The validation study indicated that it was important to combine the codes 820 and 821 in this age group. Admission rates increased over the period studied in both HIPE and ORLS datasets. In HIPE the pattern was of two plateaux separated by a period of rapid rise in the late 1970s. In the ORLS data there was a more steady rise. Statistical analysis showed significant period and cohort effects but much of this was attributable to the component of the model common to both period and cohort effects (termed "drift"). CONCLUSIONS: The finding that admission rates increased in both datasets, combining relevant codings and restricting analysis to emergency admissions, strongly suggests that the rise was real. At least part of the period effect in the HIPE data, however, might be attributable to a sampling artefact. The cohort effect in incidence rates of femoral fracture has not been previously shown and would be compatible with a number of aetiological hypotheses. PMID- 9328552 TI - Suicide in the 12 months after discharge from psychiatric inpatient care, Scotland 1968-92. AB - STUDY OBJECTIVE: To investigate the rate of suicide in the 12 months after discharge from psychiatric hospital and to determine its relationship to age, diagnosis, and period. DESIGN: Cohort study of patients discharged from psychiatric hospital. SETTING: Scotland. PARTICIPANTS: Altogether 159,742 men and 178,271 women, aged 15-84, who were discharged from Scottish psychiatric hospitals during 1968-92. MAIN RESULTS: During the 25 year period, 1212 male patients committed suicide in 198,059 person years at risk (612 per 100,000; 95% confidence interval (CI) 578,647) and 1099 female patients committed suicide in 228,993 person years at risk (480 per 100,000; 95% CI 452, 509). The overall standardised mortality ratio (general population rate = 1) was 27 (95% CI 26, 29) in men and 40 (95% CI 38, 43) in women. There were variations in the suicide rates in relation to age, diagnosis, and period. The ratio of the 1-28 day rate to the rate between days 29 and 365 over the whole study period was 1.7 (95% CI 1.4, 1.9) in men and 1.6 (95% CI 1.3, 1.8) in women. CONCLUSIONS: The variations in the post discharge suicide rate by age, sex, diagnosis, geographical location, and period suggest that there are several risk factors which vary in their distribution. Further study of these may lead to the development of effective interventions. PMID- 9328553 TI - Fatal methadone and heroin overdoses: time trends in England and Wales. AB - STUDY OBJECTIVE: Although the total number of self poisonings in England and Wales has dropped by 32%, the number involving methadone and/or heroin rose by 900% in 1974-92. Because of concern about the role of methadone in this increase, the part played by methadone and heroin in poisoning deaths in England and Wales in 1974-92 was investigated. DESIGN: A proportional mortality design was used to study whether the ratio between deaths involving methadone or heroin and other substances had increased. The time trend was examined with logistic regression. SETTING: England and Wales, 1974-92. SUBJECTS: Accidental, undetermined, and suicidal poisoning deaths (n = 43,231). MAIN RESULTS: The proportions of poisoning deaths involving methadone (alone or in combination with heroin) rose by 80% (95% CI 69%, 92%) per 3 year period. The proportion of poisoning deaths involving heroin without methadone rose by 76% (95% CI 60%, 93%) per 3 year period. Similar results were obtained when poisoning deaths were examined in relation to gender and legal category (suicide and undetermined versus accidental deaths). CONCLUSIONS: The impact of opiate addiction on rates of death by poisoning is rising quickly. This may reflect the growth of the addict population and is an important public health problem. There is no evidence that methadone's involvement in these deaths has risen disproportionately in relation to that of heroin up to 1992. PMID- 9328554 TI - Audit of ascertainment of deaths to children born in Cumbria, UK, 1950-89 through the NHS central register. AB - STUDY OBJECTIVE: To evaluate the completeness of notification of deaths by the National Health Service Central Register (NHSCR) for England and Wales. DESIGN: Deaths for a birth cohort were ascertained through scanning the relevant volumes of NHSCR. Attempts were made to confirm these deaths and additional deaths were ascertained through searching local records. Logistic regression was used to investigate how the probability of a death being missed by NHSCR varied with the year of birth, age at death, sex, and social class. SETTING: Deaths up to the end of 1989 in the CA postal area among 264,046 children born between 1950 and 1989 to mothers living in Cumbria. RESULTS: NHSCR originally ascertained 4139 deaths; local searches confirmed 3338 (81%) of these and found an additional 342. Most deaths missed by the NHSCR were neonatal deaths in the 1950s and 1960s. In the 1950s, 31% of children who died in the neonatal period either were not entered on NHSCR or, if they were entered, there was no record of their death. For children born from 1970 onwards, ascertainment of deaths through NHSCR was over 99% complete. CONCLUSIONS: The NHSCR was started in 1948 for the administration of records of National Health Service patients. It seems that many babies who died soon after birth were not therefore recorded. In parallel with the increasing use of NHSCR for epidemiological purposes, there has been a substantial and continuing improvement in its clerical procedures since the mid 1960's. PMID- 9328555 TI - Poliomyelitis trends in Pondicherry, south India, 1989-91. AB - STUDY OBJECTIVES: To assess the poliomyelitis trend, including study of the epidemiological features, and to correlate this with the immunisation coverage of infants. DESIGN: Three annual lameness surveys in children aged 0-60 months employing cluster sampling methods and a series of five cross sectional surveys of immunisation coverage in children aged 12-23 months of age were undertaken. SETTING: Pondicherry, India, 1988-92. SUBJECTS: More than 10,000 children in the age group of 0-60 months took part in the three annual lameness surveys and samples of 210 children aged 12-23 months were covered each year in immunisation coverage surveys. MEASUREMENTS AND MAIN RESULTS: Altogether 50 of 11,461, 24 of 10,093, and 17 of 11,218 children surveyed during 1989, 1990, and 1991 respectively had become lame as a result of poliomyelitis, giving prevalences of 4.4, 2.4, and 1.5 per 1000 children for the three surveys. The corrected prevalences of poliomyelitis were 5.9, 3.2, and 2.0 per 1000 children during 1989, 1990, and 1991 respectively. The proportion of cases aged up to 36 months fell from 48% in 1989 to 12.5% in 1990 and 6% in 1991. The age at onset was less than 1 year in most. The median age at onset was 10.7 months. About 54% of the affected children had received three doses of oral poliomyelitis vaccine (OPV) before the onset of paralysis. In 1988 immunisation coverage for the third dose of OPV was 91% and in 1992 it was 97.6%. The drop out rate for the first versus the third dose of OPV fell from 6.3 in 1988 to 1.9% in 1992. CONCLUSION: Three successive annual lameness surveys showed that poliomyelitis was declining between 1989 and 1991. Five immunisation coverage surveys conducted from 1988 to 1992 showed high initial coverage followed by an improvement in the form of almost universal coverage for OPV. PMID- 9328556 TI - Risk factors for seromucinous benign ovarian cysts in northern Italy. AB - STUDY OBJECTIVE: To analyse risk factors for seromucinous benign ovarian cysts. DESIGN: Between 1984 and 1994 a case-control study was carried out. Cases were 225 women aged less than 65 year with a histologically confirmed diagnosis of benign seromucinous ovarian cysts admitted to a network of obstetrics and gynaecology departments in Milan. Controls were a random sample of 450 women admitted for acute conditions that were not gynaecological, hormonal or neoplastic. They were interviewed within the framework of a case-control study of female genital neoplasms. SETTING: Network of hospitals in the greater Milan area, Italy. MAIN RESULTS: The risk of seromucinous benign ovarian cysts was higher in more educated women than in women with fewer than seven years of schooling. The odds ratios (OR) for seromucinous ovarian cysts were 1.3 and 1.4 respectively in women reporting 7-11 and > or = 12 years of schooling, and the trend in risk was statistically significant (chi(2)1 trend 5.20, p < 0.05). There was no clear relationship between the risk of seromucinous ovarian cysts and marital status, age at first marriage, oral contraceptive use, smoking or body mass index. In comparison with women reporting menstrual cycles lasting < 28 days, the risks of seromucinous cysts were 1.6, 2.6, and 2.5 respectively in women reporting cycles lasting 28-30, > or = 31 days, or with totally irregular ones. Among ever married women, nine cases and two controls reported difficulty in conception, and the corresponding OR for seromucinous cysts was 17.7 (95% confidence interval 4.2, 83.8). CONCLUSIONS: The risk of seromucinous benign ovarian tumours is greater in more educated women and in women with a history of infertility and with long or irregular menstrual cycles. PMID- 9328557 TI - Results of a pilot study of endoscopic screening of first degree relatives of colorectal cancer patients in Italy. AB - STUDY OBJECTIVE: Screening recommendations for colorectal cancer include sigmoidoscopy in asymptomatic, average risk persons aged 50 and over and colonoscopy every three to five years in high risk groups. Little is known about the eligible population's compliance with endoscopic screening. This is the first Italian report of an endoscopic screening programme for colorectal cancer patients' relatives. DESIGN: In 1986, a pilot project for colorectal cancer screening by endoscopy in high risk subjects was started in the Desio (Milan, Italy) public health service region. The results obtained after seven years are described. SETTING: The names of 536 inhabitants with colorectal cancer diagnosed between January 1975 and December 1984 and their relatives' addresses were obtained from the Regione Lombardia Health System records and from the municipal registry offices respectively. PARTICIPANTS: From October 1986 to October 1993, 778 first degree relatives aged 20-75 were offered colonoscopy. MAIN RESULTS: After seven years, 233 (29.9%) had undergone endoscopic examination, mostly up to the splenic flexure. Being > 60 in age at the start of the programme negatively affected the participation (p < 0.05). Two cancers were detected and adenomatous polyps were found in another 24 of those screened (frequencies 0.9% and 10.3% respectively). Male gender (p < 0.05), increasing age in males (p < 0.01), and two or more affected relatives in females (p < 0.01) positively affected the frequency of polyps detection. CONCLUSION: These results suggest that about 30% of the eligible population would comply at least with sigmoidoscopic screening. The collaboration of family doctors and more widespread public information about the ability to cure colorectal cancer are necessary for better compliance. PMID- 9328558 TI - Does GP fundholding affect the use of tertiary services in the UK? PMID- 9328559 TI - Coffee consumption and coronary heart disease mortality in Scottish men: a 21 year follow up study. PMID- 9328560 TI - Cost effectiveness of a prize draw on response to a postal questionnaire: results of a randomised trial among orthopaedic outpatients in Edinburgh. PMID- 9328561 TI - Induced abortion as an independent risk factor for breast cancer. PMID- 9328562 TI - RNAse protection assays for the simultaneous and semiquantitative analysis of multiple murine matrix metalloproteinase (MMP) and MMP inhibitor mRNAs. AB - Matrix metalloproteinases (MMPs) are a family of proteinases that play a major role in the metabolic degradation of extracellular matrix proteins. In order to examine the expression pattern of different MMP or MMP-inhibitor genes two RNase protection assays (RPAs) were developed that allow the simultaneous and semiquantitative assessment of their respective mRNAs. Probes for the detection of MMPs stromelysin 1, 2 and 3, matrilysin, metalloelastase, gelatinase A and B, collagenase and membrane type MMP (MT1-MMP) were included in the first RPA probe set, while probes for tissue inhibitor of matrix metalloproteinase (TIMP) 1, 2, 3 and alpha 2-macroglobulin (alpha 2-M) were included in the second probe set (inhibitor of matrix metalloproteinase-IMP set). Titration experiments revealed that this method allows the detection of MMP and inhibitor mRNAs present in at least 0.03 microgram of spleen poly(A)+ RNA. Both RPA sets were further evaluated by analyzing the expression of MMP and IMP genes in brain, kidney, spleen and liver in a murine model for endotoxemia after intraperitoneal LPS injection. Control animals showed an organ-specific constitutive expression of one or more MMPs and a high expression of TIMPs. Following LPS injection, an organ-specific upregulation or induction of MMP and TIMP RNA species was found. This change was most pronounced in the spleen, while liver, kidney and brain showed minor or no changes in MMP expression. An IMP upregulation was detected in all organs. These RPA probe sets provide a valuable tool for the simultaneous assessment of MMP and IMP gene expression under physiological and pathological conditions. PMID- 9328563 TI - Detection of colorectal cancer K-ras mutations using a simplified oligonucleotide ligation assay. AB - It has been suggested that some mutations in codons 12 and 13 of the K-ras gene are associated with the progression of colorectal adenomas to carcinomas. The aim of this study was to develop a rapid, colorimetric assay for K-ras point mutations commonly associated with colorectal cancer. K-ras exon 1 was amplified from colorectal tumor DNA and K-ras activating mutations detected using an oligonucleotide ligation assay (OLA) in combination with immunological and colorimetric detection. Using the OLA with oligonucleotides specific to individual K-ras mutations, 6 (of 17 total colorectal adenomas/carcinomas) were found to have K-ras mutations. The assay could detect as little as 10% mutant allele. A simplified OLA designed to test for either the presence (+) or absence (-) of any of the K-ras activating mutations was developed. The assay was further streamlined by use of a dipstick methodology for colorimetric development. If required, assay sensitivity can be increased by the use of the recently described EDNA-ELCA detection system. The simplified (+/-) mutation OLA in combination with a dipstick or EDNA-ELCA detection system provides a rapid, sensitive assay for K ras point mutations suitable for use as part of the clinical assessment of colorectal cancer. PMID- 9328564 TI - Improvement of Herpesvirus saimiri T cell immortalization procedure to generate multiple CD4+ T-cell clones from peripheral blood lymphocytes of AIDS patients. AB - Herpesvirus saimiri (HVS) can infect and immortalize human T lymphocytes of both CD4- and CD8-positive phenotypes. We have previously shown that infection of peripheral blood lymphocytes (PBL) from AIDS patients with HVS predominantly yielded immortalized CD8-positive T cell clones. Here we show that CD4-positive T cells from AIDS patients can be efficiently immortalized by HVS if patient PBL are enriched for CD4-positive T cell subpopulation prior to HVS infection. Such cells can be cloned and maintained in culture for prolonged times, and they exhibit activated T cell phenotype of Th1 class and are susceptible to HIV-1 infection. Several immortalized T cell clones obtained from one out of three AIDS patients tested here were HIV-1 positive and produced infectious virus. This approach permits efficient generation of multiple CD4-positive T cell clones from AIDS patients for functional and virological studies. PMID- 9328565 TI - A highly sensitive electrochemiluminescence immunoassay for interferon alfa-2b in human serum. AB - A highly sensitive immunoassay for the quantitative measurement of recombinant interferon alfa-2b (IFN alfa-2b) in serum was developed using the ORIGEN electrochemiluminescence (ECL) detection system. The ECL assay developed uses a ruthenylated mouse monoclonal anti-IFN-alfa antibody and a sheep polyclonal anti IFN-alfa antibody that has been biotinylated. An immune complex, formed between the antibodies and the IFN in the serum, is captured by streptavidin coated paramagnetic beads. The assay is sensitive and can accurately measure 4 IU/ml in undiluted serum samples. In addition, this assay requires less labor than classical ELISAs and is not significantly affected by individual serum factors. The total assay variance for both intra- and inter-assay precision is < 10%. The assay is specific for the analyte. Mean percent recoveries in pooled and individual donor serum samples range from 84 to 114%. In addition, results of the ECL assay correlate well with a bioassay and were more accurate that an ELISA for the detection of interferon alfa-2b in individual human serum samples. The assay can be modified to measure other forms of the analyte and/or interferon in other matrices. PMID- 9328566 TI - A simple high yield procedure for purification of human proteinase 3, the main molecular target of cANCA. AB - Proteinase 3, the antigen commonly recognized by classical anti-neutrophil cytoplasmic antibodies (cANCA) in patients with Wegener's granulomatosis was purified from neutrophil azurophilic granules. Proteinase 3, a serine protease with an apparent molecular mass of 29 kDa, was extracted with Triton X-100 from the azurophilic granule fraction of neutrophils after nitrogen bomb cavitation and Percoll gradient centrifugation. Anion exchange chromatography removed many proteins, which were bound to the column. The unbound proteins, which contained most of the proteinase 3, were then separated by gel filtration. All chromatography steps were done in the presence of detergent. SDS polyacrylamide gel electrophoresis of this preparation only revealed three bands migrating closely together at the position of a 29-31 kDa protein, characteristic of proteinase 3. Affinity-purified polyclonal antibodies raised against proteinase 3 were used for immunoblotting studies and demonstrated that the purified protein was proteinase 3. Antibodies to elastase, cathepsin G, myeloperoxidase, lactoferrin or lysozyme did not react in ELISA assays with the isolated protein. The proteinase 3 prepared by this procedure was found to be suitable as an antigen for detecting PR3-ANCA both in ELISA and in immunoblotting experiments. PMID- 9328567 TI - Epitope mapping by phage display: random versus gene-fragment libraries. AB - We present a comparative study on epitope mapping of four monoclonal antibodies directed against four different antigens using alternative phage display techniques and peptide scanning: mAb215 reacts with the largest subunit of RNA polymerase II, mAbBp53-11 with the tumor suppressor protein p53, mAbGDO5 with the Hantaan virus glycoprotein G2 and mAbL13F3 with the Hantaan virus nucleocapsid protein. Epitopes were determined (i) by gene-fragment phage display libraries, constructed by DNaseI digested random gene fragments cloned into the 5' terminus of the pIII-gene of fd phage and (ii) by random peptide phage libraries displaying 6mer and 15mer peptides at the N-terminus of the pIII protein. Using the gene-fragment phage display libraries a single round of affinity selection resulted in the determination of the corresponding epitopes for all monoclonal antibodies tested. In contrast, biopanning of 6mer and 15mer random peptide libraries was only successful for two of the antibodies (mAbBp53-11 and mAbGDO5) after three or four rounds of selection. For the anti-p53 antibody we recovered the epitope from both the 6mer and 15mer library, for mAbGDO5 only the 6mer library displayed the epitope sequence. However, screening of the random peptide libraries with mAb215 and mAbL13F3 failed to yield immunopositive clones. Fine mapping of the epitopes by peptide scan revealed that the minimal epitopes recognized by mAbBp53-11 and mAbGDO5 consist of four and five amino acids, respectively, whereas mAb215 requires a minimal epitope of 11 amino acids for antigen recognition. In contrast, mAbL13F3 did not react with any of the synthesized 15mer peptides. The limits of the different methods of epitope mapping tested in this study are compared and the advantages of the gene-fragment phage display system are discussed. PMID- 9328568 TI - An ELISA for hCAP-18, the cathelicidin present in human neutrophils and plasma. AB - hCAP-18 is a newly described protein of human neutrophilic granulocytes which belongs to the cathelicidin family of antimicrobial proteins. Members of this protein family share a common N-terminal sequence followed by a highly diverse antimicrobial, cationic C-terminus. The present work describes the production of recombinant hCAP-18, the generation of antibodies to the protein and the development of an accurate, sensitive and specific ELISA for the detection of hCAP-18 in cells, plasma and urine with a detection limit of 0.084 ng/ml. The amount of hCAP-18 in neutrophils is 0.627 microgram protein per 10(6) cells. The plasma level is 1.18 micrograms/ml which is several fold higher than for other neutrophil specific granule proteins. hCAP-18 is present in plasma as high molecular weight complexes. In accordance with this, hCAP-18 is barely excreted in the urine. The bone marrow appears to be the major source of plasma hCAP-18. The high level of hCAP-18 in plasma may provide an important defense against microorganisms and endotoxins. PMID- 9328569 TI - Changes in the polymorphonuclear leukocyte function of blood samples induced by storage time, temperature and agitation. AB - We have investigated changes in polymorphonuclear leukocyte (PMN) functions of blood samples caused by such typical elements of laboratory handling as storage time, temperature and agitation. The blood of five healthy subjects was stored upright in test tubes at 4, 22 and 37 degrees C over periods of 20 min, one, two, six and 24 h. Controlled agitation was performed on a shaker. The following PMN functional parameters were measured: the white blood cell count (WBC), migration, elastase (EL) release, reactive oxygen species (ROS) production and lipid peroxidation. Migration was determined in a whole-blood membrane filter assay; ROS production by latex-stimulated, luminol-enhanced chemiluminescence (CL) in a whole-blood assay; EL as EL alpha 1-antitrypsin complex; and lipid peroxidation by malondialdehyde (MDA) generation. The reactions after handling were compared with the values measured immediately after blood withdrawal which served as reference values of 'genuine' PMN reactivity. The outstanding result was the marked scatter between the individual reactions. Overall, the proportion of migrating PMNs in the blood total decreased, while CL, correlating positively with MDA, increased with the time of storage. EL increased considerably in some of the samples. Agitation raised CL and MDA. The effect of temperature was apparent only after 24 h at 37 degrees C. There was evidence that inhomogeneities in the blood samples were another interfering factor, since resuspension of sedimented blood after storage can be incomplete. In order to obtain reliable results from PMN functional tests, whole-blood assays and processing of blood samples within 20 min after blood withdrawal are recommended. PMID- 9328570 TI - Isolation of cell surface-specific human monoclonal antibodies using phage display and magnetically-activated cell sorting: applications in immunohematology. AB - A method is described for the isolation of filamentous phage-displayed human monoclonal antibodies directed at unpurifiable cell surface-expressed molecules. To optimize the capture of antigen-specific phage and minimize the binding of irrelevant phage antibodies, a simultaneous positive and negative selection strategy is employed. Cells bearing the antigen of interest are pre-coated with magnetic beads and diluted into an excess of unmodified antigen-negative cells. Following incubation of the cell admixture with a Fab/phage library, the antigen positive cell population is retrieved using magnetically-activated cell sorting and antigen-specific Fab/phage are eluted and propagated in bacterial culture. Utilizing this protocol with magnetically-labeled Rh(D)-positive and excess unlabeled Rh(D)-negative human red blood cells and a Fab/phage library constructed from human peripheral blood lymphocytes, dozens of unique clinically useful gamma 1 kappa and gamma 1 lambda anti-Rh(D) antibodies were isolated from a single alloimmunized individual. This cell-surface selection method is readily adaptable for use in other systems, such as for the identification of putative tumor-specific antigens and provides a rapid (< 1 month), high-yield approach for isolating self-replicative antibody reagents directed at novel or conformationally-dependent cell-surface epitopes. PMID- 9328571 TI - Detection of blood group antigens utilising immobilised antibodies and surface plasmon resonance. AB - Surface plasmon resonance (SPR) detection using the BIAcore biosensing system was employed for the detection of blood group-associated antigens (BGAA) on whole erythrocytes. The quantitative detection of erythrocytes was accomplished by covalently immobilising blood group-specific antibodies (IgM) to a dextran matrix and monitoring the cell binding response. Non-specific binding of erythrocytes to the IgM coated surface was not detected. Relatively mild regeneration conditions (20 mM NaOH) were employed to elute bound erythrocytes in order to preserve the activity of the immobilised antibody and allow the surface to be used repeatedly. Regeneration of the surface was particularly difficult when a high IgM immobilisation level was used and when the number of bound cells was high. Despite these considerations, a quantitative relationship between the cell binding response and erythrocyte concentration was confirmed. Erythrocyte preparations, diluted by a factor of ten as compared to physiological concentrations, were detectable. The occurrence of non-specific false positives appears to be minimal and allows the system to be used for blood typing. As a model study, the lectin concanavalin A (ConA) was covalently immobilised onto a hydrophilic dextran matrix and successfully used to support the capture of erythrocytes from suspension. PMID- 9328572 TI - Modulation of the antigenic reactivity of the citrus tristeza virus coat protein. AB - Trapping properties of a panel of monoclonal antibodies (Mabs) raised against citrus tristeza virus (CTV) were analyzed in an indirect double-antibody sandwich ELISA (I-DAS-ELISA). These antibodies had been previously assigned by serological specificity into five groups (I to V). Mabs from group V, which are directed to conformational epitopes, trapped significant amounts of virus antigen from CTV infected plant tissue at IgG concentration above 10 ng/ml. Mabs from groups I to IV, which are directed to linear, continuous epitopes, performed poorly as coating antibodies, even at a 1 microgram/ml concentration of the IgG's, indicating that the respective linear epitopes were inaccessible. However, when Mabs from groups I to IV were combined with a small amount of Mabs from group V, a substantial increase in trapping of the CTV antigen was recorded. In this 'two antibody-binding assay' previously cryptic, linear epitopes of the CTV CP apparently became accessible to the Mabs from groups I to IV. Modulation of the antigenic reactivity of the CTV CP was also recorded upon binding of the Mabs directed to the conformational epitopes in solution. Induced exposure of the linear epitopes of the CTV CP was revealed in 'two antibody-binding assays' with pairwise combinations of different mouse Mabs and several rabbit and chicken polyclonal antisera with different serological specificities, including antisera to bacterially expressed CP fragments. This mixed coating in I-DAS-ELISA resulted in substantially increased efficiency of the virus antigen trapping by antisera produced against bacterially expressed protein fragments and an increased sensitivity of the CTV detection after optimization of the ratio between conformational and linear antibodies. PMID- 9328574 TI - Very low density lipoproteins and interleukin 2 enhance the immunogenicity of 9-O acetyl-GD3 ganglioside in BALB/c mice. AB - Gangliosides expressed by tumor cells constitute potential targets for immunotherapy. A major limitation of protocols aiming to immunize patients against tumor gangliosides is the weak immunogenicity of these molecules. We have previously shown that exogenous gangliosides are essentially bound to serum lipoproteins. In this study we have analyzed the influence of human serum lipoproteins on the immunogenicity of purified human ganglioside 9-O-acetyl-GD3 in BALB/c mice. Although expressed at very low levels in mice, this ganglioside was not immunogenic when administered in the form of micelles. However 9-O-acetyl GD3 adsorbed onto Very Low Density Lipoproteins (VLDL) was strongly and reproducibly immunogenic, inducing both an IgM and an IgG response, with higher titers than those obtained with total serum. The IgM antibody response appeared after a single injection whereas the IgG response was observed after 3 weeks but was stronger and more durable. The antibody response to 9-O-acetyl-GD3 bound to other serum fractions was weak or absent. The addition of recombinant interleukin 2 (IL-2) enhanced weak antibody responses to 9-O-acetyl-GD3 thereby facilitating responses to ganglioside in micelles and in protein-free Very Low Density Particles. Using in vitro assays, we demonstrated that VLDL-bound ganglioside 14C GM3 was more sensitive to the effect of neuraminidase than gangliosides bound to other lipoprotein fractions, suggesting greater accessibility of VLDL-bound gangliosides. These results indicate that VLDL-bound gangliosides are the most immunologically active fraction of serum gangliosides. VLDL or similar particles and recombinant IL-2 may be useful adjuvants for immunization with gangliosides. PMID- 9328573 TI - Characterization of anti-human interleukin-18 (IL-18)/interferon-gamma-inducing factor (IGIF) monoclonal antibodies and their application in the measurement of human IL-18 by ELISA. AB - Interleukin-18 (IL-18)/interferon-gamma-inducing factor (IGIF) is a novel cytokine, which is a potent inducer of IFN-gamma production and plays an important role in Th1 responses. In order to develop a specific ELISA for the measurement of human IL-18, we established 13 anti-human IL-18 monoclonal antibodies and characterized them. 7 murine anti-human IL-18 mAbs and 6 rat anti human IL-18 mAbs were obtained by fusion of splenocytes from mice or rats immunized with human IL-18, with SP2/0 myeloma cells. These antibodies were classified into 4 groups according to competitive binding ELISAs to the human IL 18 molecule. 1 murine mAb and all 6 rat mAbs neutralized IFN-gamma production induced by IL-18. A specific human IL-18 ELISA was developed using two neutralizing mAbs (#125-2H and #159-12B). This ELISA detects human IL-18 with a minimum detection limit of 10 pg/ml, but does not react with heat-denatured human IL-18. The ELISA does not show any cross-reactivity with other cytokines. Using this assay, human IL-18 was measurable in the plasma of leukemia patients. This ELISA would become a powerful tool for investigating the relationship between IL 18 and various diseases or analyzing the control mechanisms of IL-18 production from IL-18 producing cells. PMID- 9328575 TI - Conjugation to preadsorbed preactivated proteins and efficient generation of anti peptide antibodies. AB - A solid phase conjugation method is described based on the preadsorption of proteins to aluminium hydroxide adjuvant followed by activation of the adsorbed carrier proteins with iodoacetic acid N-hydroxysuccinimidester or other conjugation reagents. Cysteine-containing peptides were coupled to the iodoacetic acid-activated carrier-adjuvant particles through their SH groups. No dialysis is required since the reaction product is isolated at each step of the procedure by a simple centrifugation and can easily be extensively washed between individual manipulations. The method generates peptide-carrier-adjuvant particles with sterically defined presentation of the peptides at the surface of the particles. When used for immunization of mice and rabbits the conjugates elicited high titered specific anti-peptide sera, which reacted well with the parent protein in ELISA. The strongest reactions were with the denatured form of the parent protein. On immunoblots antisera to the N- and C-terminus of calreticulin recognized the same M, 52,000 protein. PMID- 9328576 TI - Regression analysis of simulated radio-ligand equilibrium experiments using seven different mathematical models. AB - The objective of this study was to investigate the conditions for regression analysis of data from equilibrium experiments. One important issue was to recognize that Kd and the binding site concentration (A) are not of equal nature, although both are parameters in the regression analysis. Whereas Kd approximates to a true constant, A is subject to experimental variation due to pipetting errors and in solid-phase experiments also to uneven coating properties. While recognizing that the ideal assumptions for ordinary regression analysis are poorly satisfied, different regression models were evaluated by extensive simulations. It was first established by a 'worst case' investigation that a limited error (8%) in the dependent variable is not critical for the results obtained at curve-fitting to Langmuir's equation. Seven different equations were compared for the calculation of data representing a solid-phase equilibrium experiment with statistical but no systematic errors. All the equations are rearrangements of the law of mass action. In this setting the Scatchrd plot gave the best result, but also the double reciprocal and the Woolf plots worked well in weighted analysis. Langmuir's equation gave the best result of the 4 nonlinear regression models tested. The influence of one type of systematic error was also investigated. This assumed that 10% of the label was positioned on particles other than the functional ligand molecules. This systematic error was amplified, which resulted in a substantial bias. The calculated Kd-values varied slightly with the regression method used and were almost 24% too high in the best methods. PMID- 9328577 TI - CTL responses induced by a single immunization with peptide encapsulated in biodegradable microparticles. AB - A synthetic peptide representing a measles virus (MV) cytotoxic T cell epitope (CTL) when encapsulated in poly (D,L-lactide co-glycolide) (PLG) 50:50 microparticles induced a strong CTL response after a single intraperitoneal immunization of mice which was greater than that following administration of the peptide in Freund's complete adjuvant. A 100 micrograms dose of encapsulated peptide was shown to be more effective for CTL priming than 50 and 25 micrograms doses. A vaccine formulation prepared by simply mixing empty 50:50 PLG microparticles with the peptide resulted in the induction of CTL responses comparable to those induced by the encapsulated peptide. Moreover, a CTL response against MV-infected target cells was observed. These findings highlight the potential immunostimulatory effect of PLG microparticles for the induction of MV and peptide-specific CTL responses. PMID- 9328578 TI - Induction and detection of apoptosis in human periphery blood T-cells. AB - Freshly isolated, human peripheral blood T (PBT) cells are resistant to induction of apoptosis. In this study, however, we have shown that although small numbers of monocytes (Mo) are required for PBT cells to proliferate optimally in response to mitogenic challenge, a relatively higher percentage of Mo results in a significant decrease in PHA-, but not ConA-induced T-cell proliferation. Interestingly, the decrease in T-cell proliferation correlated to an increase in apoptotic cell death. Moreover, ConA-induced PBT-cells underwent apoptosis in the presence of PHA-pretreated Mo, suggesting a key role of monocyte activation in this system. This apoptosis-promoting effect of activated Mo appeared to depend on contact or close proximity between Mo and PBT-cells, rather than via soluble mediators. Despite an increase in apoptosis by the presence of high numbers of Mo, PHA-stimulated PBT-cells released IL-2 at elevated levels proportional to the increasing numbers of Mo in cultures. They also expressed activation marker CD69 and the IL-2R-gamma chain on the cell surface at comparable or higher levels in the presence of high versus low numbers of Mo. These data suggest that PBT-cells can embark on a normal early phase of activation prior to undergoing apoptosis, thereby providing a model system to study how T-cells are committed to either proliferation or activation-induced apoptosis. PMID- 9328579 TI - High-level production of a secreted, heterodimeric alpha beta murine T-cell receptor in Escherichia coli. AB - For structural studies, high-level production of properly folded, disulfide linked, unglycosylated protein in E. coli is an attractive alternative to production in eukaryotic systems. We describe here the production of heterodimeric, murine D10 T-cell receptor (sD10TCR) in E. coli as a secreted leucine zipper (LZ) fusion protein. Two genes, one (alpha-acid) encoding the alpha-chain variable and constant domains (V alpha and C alpha) of D10 TCR fused to an LZ 'acid' encoding sequence and the other (beta-base) encoding the beta chain variable and constant domains (V beta and C beta) fused to an LZ 'base' encoding sequence, were co-expressed from a bacteriophage T7 promoter as a dicistronic message. Secreted alpha-acid and beta-base proteins formed proper inter- and intra-chain disulfide bonds in the periplasm, bypassing the need for in vitro protein refolding. Complementary LZ sequences facilitated the formation of alpha beta heterodimers. sD10TCR-LZ was purified by affinity chromotography using a D10 TCR clonotype-specific monoclonal antibody (mAb 3D3). Typical yields of purified protein were 4-5 mg/l of culture. Purified sD10TCR-LZ was reactive with a panel of conformationally sensitive TCR-specific monoclonal antibodies, consistent with its conformational integrity and appeared to be suitable for structural studies by X-ray crystallography or NMR spectroscopy. PMID- 9328580 TI - Hierarchical affinity maturation of a phage library derived antibody for the selective removal of cytomegalovirus from plasma. AB - Recombinant antibody fragments can be produced in large quantities using bacterial expression systems and could potentially be useful for the generation of biofilters for the selective removal of viral particles from fluids. A human single chain-Fv antibody library, derived from synthetic repertoires of germ line VH-gene segments rearranged in vitro and paired to a single light chain (Nissim et al., 1994, EMBO J., 13, 692-698), has recently been used to isolate hundreds of different binding specificities by panning with antigen. Antibodies from this library typically have affinities in the 10(6)-10(7) M-1 range. Occasionally, better binders are isolated but at other times the affinities recovered are poor. In the latter situation binding cannot be detected with soluble antibodies, but only by high-avidity display of multiple copies of antibodies on phage. By panning with human cytomegalovirus (HCMV)-coated immunotubes, we have isolated a number of antibody clones from this library that bound to the antigen only if displayed on the filamentous phage, but not in soluble form. One of these clones was selected for an affinity maturation procedure, achieved by combinatorial mutagenesis of the complementarity determining region 3 (CDR3) of the antibody light chain, followed by selection of the resulting library for HCMV binding. By this means, we were able to isolate a number of binders, some of which exhibited specific HCMV binding in soluble form. The clone that gave the strongest ELISA signal was expressed in bacteria, purified in solution, characterised using a novel capture methodology with surface plasmon resonance detection on a BIAcore instrument and used for the production of an immunofilter for the removal of HCMV form human serum. The filter removed more than 99% of applied HCMV in 10 min circulation time, while the amount of HCMV retained non-specifically in a cartridge derivatised with a non-specific antibody was less than 10% under similar conditions. PMID- 9328581 TI - RDA of lymphocyte subsets. PMID- 9328582 TI - Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. AB - Cytokines are produced and function at a micro environmental level: intracellular assessment has only recently become practically feasible. We used 3-color flow cytometry to examine surface and cytoplasmic antigens on peripheral blood lymphocytes of 18 normal donors, assessing the applicability/comparability of various directly conjugated anti-human cytokine reagents and stimulation protocols using separated cells or whole blood preparations. Interdonor variability far exceeded variability due to reagent or stimulation and separation techniques. Based on all results with various reagents, post 4-5.5 h stimulation with PHA/PMA/ionomycin, the range of the percents of T lymphocytes producing various cytokines included: gamma-IFN-13.2-65.0%, IL-2-10.0-56.7%, and TNF-alpha 17.1-79.2%. Compared to CD8+ cells, CD4+ cells more often expressed IL-2 (mean 45.7% of CD4 + vs. 21.4% of CD8+ p < 0.0001), less often expressed gamma-IFN (18.5% vs. 55.3%, p < 0.0001), and did not differ in TNF-alpha expression (52.9% vs. 59.4%). Of T cells producing gamma-IFN, 64.8-100.0% also produced TNF-alpha 3.5-100.0%, IL-2. Of T cells producing IL-2, 6.0-63.9% also produced gamma-IFN and 37.6-100.0%, TNF-alpha. These results demonstrate the broad spectrum of cytokine patterns in normal human adults, as well as the usefulness and limitations of various currently available cytokine products. PMID- 9328583 TI - Competitive particle concentration fluorescence immunoassays for measuring anti diabetic drug levels in mouse plasma. AB - Two competitive particle concentration fluorescence immunoassays were developed to measure blood levels of analogs of anti-diabetic drugs being tested in diabetic mice. Ligands that contained the active pharmacophores were conjugated to PPD for immunization and to beta-phycoerythrin for use as a tracer in the immunoassays. Approximately 90% of 262 compounds assayed were detectable at less than 120 nM in plasma which was well below the estimated therapeutic level of 1 microM for lowering blood glucose. These data were used to define the bioavailability of test compounds and assist in decisions of constructing active analogs. Of additional interest, we noted crossreactivity of one monoclonal antibody for 3 different compound classes that are all known to bind with varying affinities to peroxisome proliferator-activated receptors. PMID- 9328584 TI - A simple method for evaluating the rejection of grafted spleen cells by flow cytometry and tracing adoptively transferred cells by light microscopy. AB - In this report we describe a simple method using the vital dye 5- (and 6-) carboxyfluorescein diacetate succinimidyl ester (CFSE) to follow splenic graft rejection by flow cytometry. CFSE-labelled spleen cell suspensions were injected intravenously into various recipients and blood samples were taken at different time points to follow the transferred cells. We found that the labelled cells could be readily detected by flow cytometry for up to eleven weeks. The loss of these labelled cells in various transfusion experiments with different major and minor histocompatibility differences followed the rejection kinetics previously described for skin transplants. Thus, this method offers a simple tool to test the histocompatibility of donor cells/grafts with the host in adoptive/transplantation experiments in which donor and host are not completely syngeneic. Furthermore, we developed a method to trace adoptively transferred fluorescent CFSE-labelled cells by light microscopy by converting in situ immunofluorescence staining into immunoenzyme staining. PMID- 9328585 TI - A very sensitive coupled luminescent assay for cytotoxicity and complement mediated lysis. AB - The demand for convenient and sensitive means of measuring cytotoxicity and complement-mediated killing is likely to be increased by the recent identification of Complement Factor H, an important regulatory protein of both the classical and alternate pathways of complement, as a tumor-associated antigen. Here we describe a simple luminometric assay capable of detecting the death of approximately 0.03 nucleated human-cell equivalent or approximately 1 rabbit-erythrocyte equivalent. The assay measures the release of glyceraldehyde-3 phosphate dehydrogenase (G3PDH) from dead or damaged cells by coupling its enzymatic activity to production of ATP, which in turn is measured by well-known methods involving firefly luciferase. This is accomplished by means of a reaction series in which the activity of G3PDH is coupled with that of phosphoglycerate kinase, the next enzyme in the glycolytic pathway. As described, the assay uses inexpensive, commercially available reagents. This coupled assay was used to demonstrate that an anti-factor-H antibody is capable of enhancing complement mediated killing of the Raji cancer cell line by > 1000%. PMID- 9328586 TI - Generation of neutralizing antibody to the reverse transcriptase of human immunodeficiency virus type 1 by immunizing of mice with an infectious vaccinia virus recombinant. AB - Antibodies inhibiting the reverse transcriptase (RT) of human immunodeficiency virus type-1 (HIV-1) were found to be generated in the serum of mice repeatedly infected with a vaccinia virus recombinant, WRRT, expressing the enzyme. A monoclonal antibody (mAb), 7C4, which specifically and almost completely inhibits the RNA-dependent DNA polymerase activity of HIV-1 RT was produced from a mouse repeatedly immunized with WRRT. 7C4 seems to be specific for HIV-1 among retroviruses: 7C4 inhibited RT activity of three strains of HIV-1 (IIIB, Bru, and IMS-1) but not of two strains of HIV-2 (GH-1 and LAV-2) or two strains of SIV (MAC and MND). The immunoglobulin isotype of three out of four mAbs produced from spleen cells of the immunized mouse were IgG2a. This immunization method that avoids protein denaturation may preferentially induce a T helper type-1 immune response and increase the chances of producing the only occasionally obtainable mAb capable of recognizing a conformational epitope and completely inhibiting enzyme activity. PMID- 9328587 TI - A new method for the analysis and production of monoclonal antibody fragments originating from single human B cells. AB - The phage display approach has proven to be a major step forward in studies on the human autoimmune repertoire. However, it remains doubtful whether the heavy and light chains of the antibodies obtained from these libraries resemble original in vivo pairings. Here we describe a novel, simple method for the immortalization of the variable heavy and light chain regions originating from individual, nonboosted, autoantigen-specific human B cells. Our method is based on the clonal expansion of B cells in which cell-cell interactions (CD40-CD40L) as well as soluble factors were shown to be essential. This B cell culture system combined with a selection on antigen (the U1A protein, a frequent autoantigenic target in patients with systemic lupus erythematosus) and single cell sorting resulted in the isolation of U1A-specific human B cells and the subsequent expression of an U1A-specific single chain variable fragment (scFv). Our method circumvents laborious plating and screening and has the advantage that original heavy/light chain pairings can be isolated. Due to the high growth efficiency of single cultured B cells (50-70% outgrowth) even rare B cell activities can be studied using this system. PMID- 9328588 TI - A rapid streptavidin-capture ELISA specific for the detection of antibodies to feline foamy virus. AB - We report a simple procedure for the rapid development of an ELISA with the potential for wide application to any defined protein antigen. The procedure involves the expression of protein encoded by a PCR product, using a commercially available T-vector that adds a biotin tag, and a single step purification by affinity for streptavidin for direct use in ELISA. In our experiments, a recombinant protein from the nucleocapsid domain of the feline foamy virus gag gene was expressed as a fusion protein with a biotin tag and then applied directly to streptavidin-coated ELISA wells. An extract from a clone with the insert in antisense orientation was used as a control. Non-specific reactions with antigen extracts from both sense and antisense clones were observed in 6 of the 376 (1.6%) sera tested. Antibody to feline foamy virus, which forms a stable persistent infection in cats, was detected in 107 of 201 (53%) Australian cats, but none of 175 sera from veterinarians. There was a 100% correlation between FeFV antibody detected by ELISA, immunoblot, serum neutralisation and virus isolation, confirming that this test is sensitive and specific. PMID- 9328589 TI - Standardizing the immunological measurement of advanced glycation endproducts using normal human serum. AB - Advanced glycation endproducts (AGEs) have been linked to many sequelae of diabetes, renal disease and aging. To detect AGE levels in human tissues and blood samples, a competitive enzyme-linked immunosorbent assay (ELISA) has been widely used. As no consensus or standard research method for the quantitation of AGEs currently exists, nor a universally defined AGE unit available, the comparative quantitation of AGEs between research laboratories is problematic and restricts the usefulness of interlaboratory clinical data. By comparing the cross reactivities of five different anti-AGE antisera with five different in vitro AGE modified proteins, we found that the immunological recognition of AGEs by competitive ELISA is both AGE-carrier protein- and anti-AGE antibody-dependent. This suggests that in vitro AGE-modified proteins might not be appropriate standards for AGEs that occur naturally in vivo. Based on our observation that serum AGE levels in the normal human population are consistently within a narrow range and several folds lower than in diabetics, we propose a method to standardize AGE units against normal human serum (NHS). In this new method, one AGE unit is defined as the inhibition that results from 1:5 diluted NHS in the competitive AGE-ELISA; thus the AGE value in NHS is 5 units/ml. This NHS method requires a competitive AGE-ELISA with reasonable sensitivity such that 1:5 NHS produces a 25 to 40% inhibition of anti-AGE antibody binding to immobilized AGE proteins. By using this standardized method we found that the AGE levels in normal human serum (5.0 +/- 2.2 units/ml; mean +/- SD, n = 34) fit a normal distribution (chi 2-test, p < 0.01), and the serum AGE levels in diabetic patients (20.3 +/- 3.8 units/ml, n = 7) are significantly higher than that of the normal population (p < 0.0001). Since AGE units can now be defined against a universally available standard, NHS, the results of quantitative AGE measurements using this method should be comparable between assays and between different laboratories. Taken together, standardizing the AGE-ELISA protocol as described here provides a simple and quantitative method that should facilitate the expanded application of clinical AGE data. PMID- 9328607 TI - Evidence for genetic variability in venous responsiveness. AB - Alterations in adrenergic mediated vascular responsiveness and glucose metabolism have been implicated in the pathogenesis of essential hypertension. We sought for a human model to study possible interactions between the two pathogenetical factors. In a group of Latin-Americans there is a predominance of diabetes mellitus, impaired glucose tolerance and obesity, while the rate of hypertension is unexpectedly low (lower than in the normal population). Here we demonstrate attenuated alpha 1 adrenergic mediated vasoconstriction in the above group, suggesting a possible protective mechanism against the development hypertension. PMID- 9328590 TI - Antigenic features of prion proteins of sheep and of other mammalian species. AB - Pathological prion protein (PrPSc) which is a conformational isoform of a host encoded protein designated (PrPC) serves as a specific marker protein for the immunochemical diagnosis of transmissible spongiform encephalopathies (TSE). The generation of suitable antibodies to PrPSc therefore underlies the specificity and sensitivity of diagnostic assays. However, most antibodies reported to date are directed to a limited number of epitopes only. PrPC is a highly conserved cell membrane protein in all mammalian species studied to date. In an attempt to generate antibodies to further regions of PrP we raised antisera in rabbits and chicken against sixteen synthetic peptides which represent the complete aminoacid sequence of ovine PrP. By this approach immunotolerance was overcome and immunoblot-reactive antibodies were stimulated to epitopes at almost any site of ovine PrPC and PrPSc. A large number of different antibodies cross-reacted also with affinity-purified PrPCs from other mammalian species including cow, goat, pig, man, dog, cat, mink, mouse, hamster and guinea pig. No epitope, however, was recognized exclusively on the pathological or cellular isoform of PrP indicating that both isoforms occur in highly denatured conformations on the immunoblots. Antibodies to the amino-terminus are suitable for immunoprecipitation of PrP. The availability of rabbit and chicken anti-peptide antibodies to PrP will greatly improve immunochemical diagnosis and pathogenetic studies on these diseases. PMID- 9328608 TI - Effect of ACE inhibition by benazepril, enalapril and captopril on chronic and post exercise proteinuria. AB - Although post exercise proteinuria has long been known, its exact pathophysiology is unclear. Our objective was to determine whether long-term angiotensin converting enzyme (ACE) inhibition by different ACE inhibitors had an influence on post exercise proteinuria. We studied 14 patients who also had mild, chronic proteinuria caused by diabetes mellitus or chronic glomerulonephritis. We compared changes both in chronic (baseline) and post exercise proteinuria, during and after treatment with three different ACE inhibitors, with appropriate washout periods for the three drugs to all 14 patients. Proteinuria (mg/24 hours +/- SD), prior to the treatment was 682 +/- 92. Proteinuria after treatment for 30 days with benazepril was 464.4 +/- 82.6 (p < 0.001), with enalapril: 477.1 +/- 105.5 (p < 0.001), and captopril: 504.7 +/- 100.1 (p < 0.001). Proteinuria three days after discontinuing the treatment with benazepril was 532.4 +/- 113.5, (p < 0.01), with enalapril: 561.3 +/- 128.5, (p < 0.01), and with captopril: 620.8 +/- 101.8, p = n.s. Post exercise proteinuria prior to treatment (mg/min. +/- SD) was: 1.38 +/- 0.32, vs. after a 30-day treatment period with benazepril: 0.81 +/- 0.19 (p < 0.001), enalapril: 0.95 +/- 0.24, (p < 0.001), captopril: 1.09 +/- 0.27 (p < 0.02). Post exercise proteinuria three days after discontinuing the treatment was (blood pressure already back to baseline): in case of benazepril: 1.26 +/- 0.36 (p = n.s.), of enalapril: 1.17 +/- 0.46 (p = n.s.), and of captopril: 1.34 +/- 0.41 (p = n.s.). We conclude that the renin-angiotensin system plays a significant role in the pathogenesis of post exercise proteinuria; the antiproteinuric effect of ACE inhibition in exercise-induced proteinuria seems to be associated chiefly with the hemodynamic changes due to these drugs, whereas in chronic proteinuria the antiproteinuric and antihypertensive effects are, at least partially, dissociated. PMID- 9328609 TI - Xenobiotic metabolizing enzymes in fish: diversity, regulation and biomarkers for pollutant exposure. AB - Cytochromes P450 play key roles in biotransformation of pollutant chemicals and in the activation or inactivation of many toxic or carcinogenic compounds. Multiple P450 isozymes have been purified from different fish species. Fish monooxygenase activity shows temperature compensation and sex-related variation. Several xenobiotics can induce cytochrome P450 monooxygenases altering toxicity of chemical contaminants. Polycyclic aromatic hydrocarbons can increase transcription of CYP1A gene in fish as it has been observed in mammals, but phenobarbital-type agents do not induce in fish at all. The presence of conjugation enzymes in fish has also been proved, although their induction by xenobiotics is poorly investigated. Since exposure of fish to environmental contaminants can result in the induction of specific cytochrome P450 enzymes, monitoring of their catalytic activities can identify polluted areas. PMID- 9328610 TI - Effects of administration of temazepam on blood glucose and serum lipid levels in hyperlipidemic rats. AB - In rats rendered hyperlipidemic by the administration of Triton WR-1339 temazepam induced significant reductions of serum lipids. The effects was more reduced than of diazepam. The blood glucose level failed to be affected by most of the doses. PMID- 9328613 TI - Platelet rich plasma serotonin content in patients suffering from different psychiatric disorders. AB - Platelet rich plasma serotonin contents were examined using HPLC-EC method in patients suffering from different anxiety states and compared to age matched healthy controls. We have found significant increase of PRP serotonin level in the group of schizophrenic patients and in patients suffering from dementia compared to controls. PRP serotonin content was significantly lower in heavy drinkers but in patients suffering from panic disorder did not differ significantly from the controls. PMID- 9328612 TI - Studies on the sites and mechanisms of 5-HT1A receptor-mediated in vivo actions. AB - In the first study the possible role of the hypothalamic paraventricular nucleus (PVN) in 5-HT1A receptor agonist-induced neuroendocrine responses was tested. Surgical lesions of the PVN completely blocked ACTH and corticosterone and markedly attenuated oxytocin but not prolactin responses to ipsapirone, providing evidence for a crucial role of the PVN in these responses. In the second study we have compared the effectiveness of intracerebroventricular and intravenous administration of 8-OH-DPAT in frequently used behavioural models of 5-HT1A receptor activation, namely lower lip retraction, body temperature and tail flick responses. We have found marked differences in the rates of effectiveness. We conclude that these models measure the activation of different subsets with clearly separate location of 5-HT1A receptors. PMID- 9328611 TI - Effects of midazolam on glycemia and serum lipids in rats. AB - Midazolam administered ip. in albino rats (each group consisted from 10 animals rendered hyperdyslipidemic by the administration of Triton WR-1339) induced at most doses a significant reduction of glycemia (p < 0.001). However, the reduction of blood glucose level was outside of the dangerous level. Midazolam elicited also very significant decrease of the elevated serum lipids (p < 0.001). The pharmacological analysis of these phenomena by using the peripheral type benzodiazepine (BZD) receptors antagonist PK 11105, the central BZD receptor antagonist flumazenil and the purinergic P1 receptors antagonist aminophylline has shown that the effects on serum lipids were due, very probably to the stimulation of the peripheral type BZD receptors. Aminophylline seems to have the property to block the peripheral type BZD receptors. The effects on blood glucose level were very variable. PMID- 9328614 TI - Antiestrogens, antiandrogens. AB - The aim of the study was to find new antiestrogenic and antiandrogenic structures. Out of the triphenyl-alkene derivatives Panomifene (EGIS-5660) proved to be the most active antiestrogenic compound which binds to specific estrogen receptors and exhibits inhibitory effects on experimental mammary tumors both in vitro and in vivo. The investigated antiandrogenic compounds were indol and imidazole derivatives. One of these compounds a di-imidazolil derivative, GYK1 24479 inhibited the in vitro androgen (testosterone and androstenedione) biosynthesis both in vitro and in vivo in concentration/dose dependent manner, and in these respects proved to be more active than the referent ketoconazole. PMID- 9328615 TI - Electrophysiological effects of WAY 100635, a new 5-HT1A receptor antagonist, on dorsal raphe nucleus serotoninergic neurones and CA1 pyramidal cells in vitro. AB - The novel 5-HT1A receptor antagonist WAY 100635 [(N-(2-(-4(2-metoxyphenyl)-1 piperazinyl)ethyl)-N-(2-pyridinyl)cyc lohexane carboxamide)] has been tested on 5 HT1A receptor-mediated inhibition of firing and intracellularly recorded hyperpolarisation of serotoninergic cells of the dorsal raphe nucleus (DRN) and on hyperpolarisation of hippocampal CA1 pyramidal cells. WAY 100635 selectively blocked 5-HT1A receptor-mediated responses of 5-HT, 8-OH-DPAT, lesopitron and 5 CT. The antagonism of the hyperpolarisation elicited by 5-CT was competitive in the DRN and non competitive in CA1, probably because of the existence of a 5-HT1A receptor reserve in serotoninergic cells of DRN. PMID- 9328616 TI - The xanthine derivatives and cerebral blood flow (CBF) after i.v. or i.c. administration in SHR rats. AB - It was shown, that TPH after i.v. administration causes significant decrease of CBF, but IBMX and P-23 had no effects on CBF. After i.c. administration of TPH and P-23 CBF decreased, whereas IBMX i.c. injection had no effect on CBF in SHR rats. These results indicate that direct administration of TPH or P-23 into the cerebral vascular beds causes intensification of TPH activity and reveals the action of P-23. These data suggest, that intracarotid administration of xanthine derivatives can change their cerebrovascular activity. PMID- 9328617 TI - Misoprostol (cytotec) in the treatment of peripheral ischaemic disease. AB - The stable prostacyclin analogue-iloprost and prostaglandin E1 (Alprostadil) showed a beneficial effect on activated platelets and leukocytes, and thrombocyte and leukocyte vessel interaction and damaged endothelium, improving microvascular perfusion and were useful in treatment of patients with peripheral arterial disease. The 4 weeks therapy with misoprostol caused a clinical improvement in all 14 patients and resulted in vasorelaxation and showed antiplatelet and fibrinolytic effects. PMID- 9328618 TI - Differences between BDZ1 selective and non-selective GABAA/BDZ receptor ligands in discriminative stimulus and EtOH intake/preference paradigms. PMID- 9328619 TI - The hippocampus--the key structure in processing of emotional input in the central nervous system. AB - Gray's conception hypothesizes that anxiolytics act indirectly to impair the behavioural inhibition system through GABA-ergic modulation of the ascending NA and 5-HT pathways to the hippocampus. The obtained results support this theory. PMID- 9328620 TI - Pretreatment with indomethacin modulates the effects of selective adenosine receptor agonists on the contractility and effective refractory period in the guinea pig heart. AB - The effects of selective adenosine receptor agonists N6-Cyclohexyladenosine (CHA, A1-agonist) and 5'-(N-ethylcarboxamido)-adenosine (NECA, A2-agonist) on the force of contraction (Fc) and effective refractory period (ERP) of isolated guinea pig papillary muscle were measured. Additionally, the influence of potent cycloxygenase inhibitor indomethacin on above mentioned parameters as well as on the effects of CHA and NECA were studied. It was found that CHA induced negative inotropic action and a shortening of ERP, opposite to NECA and indomethacin. The pretreatment with 10 microM of indomethacin shifted to the right the CHA effects and abolished the NECA effects. PMID- 9328621 TI - On the regulation of kynurenic acid production in the rat brain slices: evidences for the involvement of metabotropic glutamate receptor. PMID- 9328622 TI - The increase in cyclic nucleotide level decreases the contractile response of gastric smooth muscle strips to galanin. PMID- 9328623 TI - Desensitisation of substance P decreases the contractile effect of M15 and Gal (1 14)-(Abu 8) SCY-I on the rat gastric fundus. AB - Tachyphylaxis to SP decreased the effect of M15 and Gal(1-14)-[Abu8]SCY-I on gastric smooth muscles, without effect on the action of Gal. These findings support our initial hypothesis: the action of M15 and Gal(1-14)-[Abu8]SCY-I on the smooth muscles may not only be due to their agonist activity at Gal receptors, but may result from a subsequent stimulation of receptors for SP and perhaps for other tachykinins as well, however, a possibility that Gal analogues release endogenous SP can not be excluded. Further studies involving a tachykinin antagonist (spantide) are in progress at the moment. [1]. PMID- 9328624 TI - Efficacy and tolerance of 400 MG bezafibrate in diabetic and hyperlipidaemic patients. AB - The authors report the results of an open clinical study using 400 mg Bezafibrate once a day. Among 25 diabetic patients (type II.) underwent a 4 weeks period with nutritional advice. Average changes from inclusion levels were -24% for total cholesterol, -56% for triglicerides, +11.9% for HDL-cholesterol, -19% for plama fibrinogen. In conclusion, bezafibrate at a daily dose of 400 mg had significant lipid-modifying properties but also exhibited a beneficial effect on other related risk factors such as fibrinogen reduction. PMID- 9328625 TI - Two animal models of anxiety--different sensitivity for anxiolytic action of GABAA receptor complex ligands. PMID- 9328627 TI - Pharmacological activity of fluoxetine. AB - Some SSRIs like fluvoxamine and zimeldine have already been investigated for their memory improving activity in humans and animals, with positive results. The purpose of this paper is to observe some activities of fluoxetine the known antidepressant on some control neuron system functions [3]. PMID- 9328626 TI - Opioid receptor affinity and analgesic activity of O- and C-glycosylated opioid peptides. AB - O- and C-glycosylation of the mu-agonist dermorphin reduced neither its mu receptor affinity in binding assay nor its agonist potency in guinea-pig ileum assay (GPI). O- and C-glycosylation of the delta-agonist deltorphin reduced its delta-receptor affinity and its agonist potency in mouse vas deferens assay (MVD). O- and C-glycosylated dermorphin, administered i.c.v. and s.c., produced long-lasting antinociception in mice and rats. The ratio between i.c.v. and s.c. antinociceptive ED50 demonstrates facilitated transport into the CNS only for the galactosil peptide. Acetylation significantly reduced penetration of glycopeptides into the CNS indicating that facilitated transport into the CNS exists, but does not depend on the glucose transporter (GLUT-1). PMID- 9328628 TI - Central serotonergic system and mechanism of anxiolytic action. AB - The results clearly indicate that the hippocampus, rather than nucleus accumbens is involved in mediating anxiolytic-like effects of the 5-HT1A receptor agonists. Furthermore, hippocampal postsynaptic 5-HT1A receptors may account for the anti emotional influence of this groups of drugs. As far as the 5-HT3 receptor antagonists are concerned, it was more difficult to localize their central anti anxiety like action. More clear and unequirocal effects could be observed after intra-accumbens, rather than after intrahippocampal injections of tropisetron and ondansetron. PMID- 9328629 TI - Influence of bilateral clamping of carotid arteries on the seizures susceptibility and central action of AOAA in mice. AB - In the experiments carried out on Albino-Swiss mice it was found, that bilateral clamping of carotid arteries (BCCA) for 30 min produce the increase of GABA content in hippocampus, striatum and frontal cortex and decrease of the seizures susceptibility to bicuculline, investigated 7 days after surgery. Moreover, BCCA modulates the action of aminooxyacetic acid (AOAA): potentiates anticonvulsive effect of low doses of AOAA (33 mg/kg) and inhibits the convulsive action of high doses. Also the potentiation of the anticonvulsive effect of diazepam, phenobarbital and valproic acid (VPA) was observed in BCCA and AOAA (CD97, 150 mg/kg) treated animals. Observed effects suggests that enhanced GABA-ergic activity and decrease of excitatory amino acid (EAA) system could be involved in these processes. PMID- 9328630 TI - L-arginine--substrate for no synthesis--its beneficial effects in therapy of patients with peripheral arterial disease: comparison with placebo-preliminary results. AB - Intravenous infusions of L-arginine (L-ARG) and placebo (saline) resulted in improvement of clinical assessments, statistically significant after L-ARG but not after saline. Results of laboratory estimations for platelet and fibrynolysis changed significantly following L-ARG infusions but not after infusions of placebo. These data indicate beneficial effects of L-ARG as a therapeutic agent in patients with peripheral arterial obliterative disease (PAOD). In these patients exogenous L-ARG can be converted to NO. PMID- 9328631 TI - Pharmacokinetics and bioavailability of stereoisomeric analogues of ifosfamide. AB - Analogues of ifosfamide (IPA): racemic bromofosfamide (+/-)-(R,S)-KM 135, racemic chlorobromofosfamide (+/-)-(R,S)-CBM 4a and levorotatory enantiomer of chlorobromofosfamide (-)-(S)-CBM 11 belong to the known group of oxazaphosphorines. Antitumor activity of those three selected compounds, investigated in the Institute of Immunology and Experimental Therapy, Polish Academy of Sciences in Wroclaw against L1210 leukemia, Lewis lung carcinoma and B16 melanoma tumor model system showed cytostatic activity higher than the referential - ifosfamide [1]. These interesting data prompted us to conduct the preclinical pharmacokinetic studies in rats. PMID- 9328632 TI - The role of supraspinal modulation of alpha-2-adrenergic receptors on nociceptive process. AB - Our findings have shown that both supraspinal activation and blockade of alpha-2 adrenergic receptors change bioelectrical response of studied brain structures on the nociceptive stimulation, among other things by the modulation of hippocampal activity. The mechanism of this effect remains to be established. PMID- 9328633 TI - Zinc and cadmium ions differently modulates A1 adenosine receptors. AB - The results suggest that: 1) Zinc ions may chelate the histidines critical for the agonist binding preventing hydrogen bonds between nonprotonated nitrogen atom of His-251 and the exocyclic N6-H in CHA or CCPA molecule and between His-278 and -OH of the ribose ring. This mechanism can explain the reduction in the number of binding sites without changing the affinity. 2) Cadmium ions may oxidize cysteine SH-groups. The redox reaction between Cd2+ and receptor thiols may result in binding of the metal into stable (di)thiol-cadmium complexes rather than in the formation of disulfide and liberation of the reduced metal. This mechanism can justify the conformational modifications of the receptor molecule producing the decrease in affinity. PMID- 9328634 TI - Paracetamol induced acute interstitial nephritis superimposed on mesangiocapillary glomerulonephritis. AB - We report the case of a young alcoholic male whose first renal biopsy disclosed mesangiocapillary glomerulonephritis. One month later he took 1.5 g paracetamol to control the fever. Soon he got hospitalized due to toxicoderma, elevated liver and renal function tests. While the liver enzymes returned to normal, uremia developed. A repeated renal biopsy revealed severe interstitial inflammation, tubular atrophy. Haemodialysis was started and he got steroids (1 mg/kg body weight). He showed considerable recovery of renal function in some weeks. The case points to the possibility that paracetamol-even in therapeutic dosage-might result in hepatic and renal damage in alcoholics. PMID- 9328635 TI - NG-nitro-L-arginine sensitizes mice to 4-aminopyridine-induced seizures. AB - We investigated the influence of NG-nitro-L-arginine (NNA), the inhibitor of nitric oxide synthese, on seizures induced by 4-aminopyridine (4-AP), the K+ channel antagonist, in mice NNA (5, 10 and 40 mg/kg, i.p.) significantly reduced the respectives CD50 of 4-AP from 9.0 to 7.6, 7.5 and 6.8 for clonic seizures, and from 9.2 to 7.7, 7.5 and 6.9 for tonic seizures and death. Lower doses of NNA (1.0 and 2.5 mg/kg) had no effect on 4-AP-induced convulsions and lethality. Our results indicate that 4-AP-induced seizures may be, at least in part, dependent on nitric oxide level. PMID- 9328636 TI - Yawning induced by apomorphine, physostigmine or pilocarpine is inhibited by electroconvulsive shock (ECS). AB - Single and repeated ECS decrease sensitivity of dopamine D2 and acetylcholine receptors as measured by yawning behavior. This reduced sensitivity is dependent from the number of applied shocks, since effects of repeated ECS disappear 5 days after the last shock but effect of single ECS disappear 24 h after the shock. PMID- 9328637 TI - The estimation of interactions between arginine-vasopressin (AVP) and NMDA receptors in memory and learning processes. AB - Arginine-vasopressin (AVP) is a neuropeptide which facilitates learning and memory processes. We examinated the participation of NMDA receptors in beneficial effects of peptide. The results of our study show that noncompetitive antagonist of NMDA receptor-MK-801 impairs the effect of AVP on the consolidation of conditioned avoidance responses and antagonist of polyamines site-arcaine reduced advantageous effect of AVP on the retrieval of memory in passive avoidance situation. PMID- 9328638 TI - The influence of 1S,3R-ACPD on central dopaminergic transmission and recognition memory. AB - 1S,3R-ACPD improved learning and memory in a passive avoidance situation. We examined its influence on recognition memory and dopaminergic transmission. The results of our study shown that 1S,3R-ACPD had not influence on recognition memory and diminished dopaminergic transmission. PMID- 9328639 TI - The role of CD45 in signal transduction. PMID- 9328640 TI - HLA class II peptide binding specificity and autoimmunity. PMID- 9328641 TI - Role of cytokines in sepsis. PMID- 9328642 TI - Role of macrophage migration inhibitory factor in the regulation of the immune response. PMID- 9328644 TI - CD8+ cells in human immunodeficiency virus type I pathogenesis: cytolytic and noncytolytic inhibition of viral replication. PMID- 9328643 TI - The intrinsic coagulation/kinin-forming cascade: assembly in plasma and cell surfaces in inflammation. PMID- 9328645 TI - Nitrate assimilation by bacteria. AB - Nitrate is a significant nitrogen source for plants and microorganisms. Recent molecular genetic analyses of representative bacterial species have revealed structural and regulatory genes responsible for the nitrate-assimilation phenotype. Together with results from physiological and biochemical studies, this information has unveiled fundamental aspects of bacterial nitrate assimilation and provides the foundation for further investigations. Well-studied genera are: the cyanobacteria, including the unicellular Synechococcus and the filamentous Anabaena; the gamma-proteobacteria Klebsiella and Azotobacter; and a Gram positive bacterium, Bacillus. Nitrate uptake in most of these groups seems to involve a periplasmic binding protein-dependent system that presumably is energized by ATP hydrolysis (ATP-binding cassette transporters). However, Bacillus may, like fungi and plants, utilize electrogenic uptake through a representative of the major facilitator superfamily of transport proteins. Nitrate reductase contains both molybdenum cofactor and an iron-sulfur cluster. Electron donors for the enzymes from cyanobacteria and Azotobacter are ferredoxin and flavodoxin, respectively, whereas the Klebsiella and Bacillus enzymes apparently accept electrons from a specific NAD(P)H-reducing subunit. These subunits share sequence similarity with the reductase components of bacterial aromatic ring-hydroxylating dehydrogenases such as toluene dioxygenase. Nitrite reductase contains sirohaem and an iron-sulfur cluster. The enzymes from cyanobacteria and plants use ferredoxin as the electron donor, whereas the larger enzymes from other bacteria and fungi contain FAD and NAD(P)H binding sites. Nevertheless, the two forms of nitrite reductase share recognizable sequence and structural similarity. Synthesis of nitrate assimilation enzymes and uptake systems is controlled by nitrogen limitation in all bacteria examined, but the relevant regulatory proteins exhibit considerable structural and mechanistic diversity in different bacterial groups. A second level of control, pathway specific induction by nitrate and nitrite in Klebsiella, involves transcription antitermination. Several issues await further experimentation, including the mechanism and energetics of nitrate uptake, the pathway(s) for nitrite uptake, the nature of electron flow during nitrate reduction, and the action of transcriptional regulatory circuits. Fundamental knowledge of nitrate assimilation physiology should also enhance the study of nitrate metabolism in soil, water and other natural environments, a challenging topic of considerable interest and importance. PMID- 9328646 TI - Physiology of carbohydrate to solvent conversion by clostridia. AB - The solvent-forming clostridia have attracted interest because of their ability to convert a range of carbohydrates to end-products such as acetone, butanol and ethanol. Polymeric substrates such as cellulose, hemicellulose and starch are degraded by extracellular enzymes. The majority of cellulolytic clostridia, typified by Clostridium thermocellum, produce a multi-enzyme cellulase complex in which the organization of components is critical for activity against the crystalline substrate. A variety of enzymes involved in degradation of hemicellulose and starch have been identified in different strains. The products of degradation, and other soluble substrates, are accumulated via membrane-bound transport systems which are generally poorly characterized. It is clear, however, that the phosphoenolpyruvate-dependent phosphotransferase system (PTS) plays a major role in solute uptake in several species. Accumulated substrates are converted by intracellular enzymes to end-products characteristic of the organism, with production of ATP to support growth. The metabolic pathways have been described, but understanding of mechanisms of regulation of metabolism is incomplete. Synthesis of extracellular enzymes and membrane-bound transport systems is commonly subject to catabolite repression in the presence of a readily metabolized source of carbon and energy. While many genes encoding cellulases, xylanases and amylases have been cloned and sequenced, little is known of control of their expression. Although the mechanism of catabolite repression in clostridia is not understood, some recent findings implicate a role for the PTS as in other low G-C Gram-positive bacteria. Emphasis has been placed on describing the mechanisms underlying the switch of C. acetobutylicum fermentations from acidogenic to solventogenic metabolism at the end of the growth phase. Factors involved include a lowered pH and accumulation of undissociated butyric acid, intracellular concentration of ATP and reduced pyridine nucleotides, nutrient limitation, and the interplay between pathways of carbon and electron flow. Genes encoding enzymes of solvent pathways have been cloned and sequenced, and their expression correlated with the pattern of end product formation in fermentations. There is evidence that the initiation of solvent formation may be subject to control mechanisms similar to other stationary-phase phenomena, including sporulation. The application of recently developed techniques for genetic manipulation of the bacterium is improving understanding of the regulatory circuits, but a complete molecular description of the control of solvent formation remains elusive. Experimental manipulation of the pathways of electron flow in other species has been shown to influence the range and yield of fermentation end-products. Acid-forming clostridia can, under appropriate conditions, be induced to form atypical solvents as products. While the mechanisms of regulation of gene expression are not at all understood, the capacity to adapt in this way further illustrates the metabolic flexibility of clostridial strains. PMID- 9328647 TI - The envelope layers of mycobacteria with reference to their pathogenicity. AB - The review discusses current knowledge of the biosynthesis, composition and arrangement of the mycobacterial envelope, describes the biological activities of the constituents and considers how these activities may be relevant to the pathology of mycobacterial disease. The envelope possesses three structural components: plasma membrane, wall and capsule. Although the major biomolecules occurring in each of these parts are known, the distribution of numerous minor substances is poorly understood; an attempt has been made to assign them to particular positions on rational grounds. The plasma membrane appears to be a typical bacterial membrane but, though vital to the mycobacterium, probably plays little part in pathological processes. The wall partly resembles a Gram-positive wall, but is unusual in having a layer of lipid (mycolate esters) which is probably arranged to form a permeability barrier to polar molecules. The capsule, whose chemical composition has only recently been recognized, consists of polysaccharide and protein with traces of lipid; the arrangement of these components is imperfectly understood. Constituents of all parts of the envelope have biological activities which may be relevant. The likely importance of these activities in the overall effect of the envelope is considered. PMID- 9328648 TI - The effects of fermentation acids on bacterial growth. AB - Anaerobic habitats often have low pH and high concentrations of fermentation acids, and these conditions can inhibit the growth of many bacteria. The toxicity of fermentation acids at low pH was traditionally explained by an uncoupling mechanism. Undissociated fermentation acids can pass across the cell membrane and dissociate in the more alkaline interior, but there is little evidence that they can act in a cyclic manner to dissipate protonmotive force. Fermentation acid dissociation in the more alkaline interior causes an accumulation of the anionic species, and this accumulation is dependent on the pH gradient (delta pH) across the membrane. Fermentation acid-resistant bacteria have low delta pH and are able to generate ATP and grow with a low intracellular pH. Escherichia coli O157:H7 is able to decrease its intracellular pH to 6.1 before growth ceases, but this modest decrease in delta pH can only partially counteract the toxic effect of fermentation anion accumulation. Fermentation acid-resistant bacteria are in most cases Gram-positive bacteria with a high intracellular potassium concentration, and even acid-sensitive bacteria like E. coli K-12 have increased potassium levels when fermentation acids are present. Intracellular potassium provides a counteraction for fermentation acid anions, and allows bacteria to tolerate even greater amounts of fermentation anions. The delta pH-mediated anion accumulation provides a mechanistic explanation for the effect of fermentation acids on microbial ecology and metabolism. PMID- 9328649 TI - Physiology and genetics of sulfur-oxidizing bacteria. AB - Reduced inorganic sulfur compounds are oxidized by members of the domains Archaea and Bacteria. These compounds are used as electron donors for anaerobic phototrophic and aerobic chemotrophic growth, and are mostly oxidized to sulfate. Different enzymes mediate the conversion of various reduced sulfur compounds. Their physiological function in sulfur oxidation is considered (i) mostly from the biochemical characterization of the enzymatic reaction, (ii) rarely from the regulation of their formation, and (iii) only in a few cases from the mutational gene inactivation and characterization of the resulting mutant phenotype. In this review the sulfur-metabolizing reactions of selected phototrophic and of chemotrophic prokaryotes are discussed. These comprise an archaeon, a cyanobacterium, green sulfur bacteria, and selected phototrophic and chemotrophic proteobacteria. The genetic systems are summarized which are presently available for these organisms, and which can be used to study the molecular basis of their dissimilatory sulfur metabolism. Two groups of thiobacteria can be distinguished: those able to grow with tetrathionate and other reduced sulfur compounds, and those unable to do so. This distinction can be made irrespective of their phototrophic or chemotrophic metabolism, neutrophilic or acidophilic nature, and may indicate a mechanism different from that of thiosulfate oxidation. However, the core enzyme for tetrathionate oxidation has not been identified so far. Several phototrophic bacteria utilize hydrogen sulfide, which is considered to be oxidized by flavocytochrome c owing to its in vitro activity. However, the function of flavocytochrome c in vivo may be different, because it is missing in other hydrogen sulfide-oxidizing bacteria, but is present in most thiosulfate oxidizing bacteria. A possible function of flavocytochrome c is discussed based on biophysical studies, and the identification of a flavocytochrome in the operon encoding enzymes involved in thiosulfate oxidation of Paracoccus denitrificans. Adenosine-5'-phosphosulfate reductase thought to function in the 'reverse' direction in different phototrophic and chemotrophic sulfur-oxidizing bacteria was analysed in Chromatium vinosum. Inactivation of the corresponding gene does not affect the sulfite-oxidizing ability of the mutant. This result questions the concept of its 'reverse' function, generally accepted for over three decades. PMID- 9328650 TI - Circadian and ultradian clock-controlled rhythms in unicellular microorganisms. AB - The time structure of a biological system is at least as intricate as its spatial structure. Whereas we have detailed information about the latter, our understanding of the former is still rudimentary. As techniques for monitoring intracellular processes continuously in single cells become more refined, it becomes increasingly evident that periodic behaviour abounds in all time domains. Circadian timekeeping dominates in natural environments. Here the free-running period is about 24 h. Circadian rhythms in eukaryotes and prokaryotes allow predictive matching of intracellular states with environmental changes during the daily cycles. Unicellular organisms provide excellent systems for the study of these phenomena, which pervade all higher life forms. Intracellular timekeeping is essential. The presence of a temperature-compensated oscillator provides such a timer. The coupled outputs (epigenetic oscillations) of this ultradian clock constitute a special class of ultradian rhythm. These are undamped and endogenously driven by a device which shows biochemical properties characteristic of transcriptional and translational elements. Energy-yielding processes, protein turnover, motility and the timing of the cell-division cycle processes are all controlled by the ultradian clock. Different periods characterize different species, and this indicates a genetic determinant. Periods range from 30 min to 4 h. Mechanisms of clock control are being elucidated; it is becoming evident that many different control circuits can provide these functions. PMID- 9328651 TI - Biodegradation and metabolism of unusual carbon compounds by anoxygenic phototrophic bacteria. AB - Anoxygenic phototrophic bacteria play an important role in anaerobic nutritional cycles. The most readily used and widely studied carbon sources for growth of these bacteria are organic acids and a few carbohydrates. In this review we survey the growing knowledge on the metabolism of a number of other carbon sources, particularly polymers (starch, poly(3-hydroxyalkanoates)), aromatic compounds (natural and xenobiotic), one-carbon compounds, alcohols, aliphatic hydrocarbons and higher fatty acids, and their influence on various cellular activities of purple non-sulfur bacteria. We also discuss the possible exploitations in various biotechnological processes of this group of microorganisms while metabolizing unusual carbon compounds. PMID- 9328652 TI - Assessment of current techniques for determining tracheal luminal stenosis in dogs. AB - OBJECTIVE: To assess the accuracy of current antemortem and postmortem techniques for determining tracheal luminal stenosis. ANIMALS: 15 dogs. PROCEDURE: Percentage of tracheal luminal stenosis (PTLS) was determined by 6 methods, using measurements obtained by radiography, tracheoscopy, and necropsy after selected tracheostomy techniques were performed. To calculate PTLS, dorsoventral tracheal diameter was measured from preoperative and postoperative lateral cervical radiographic views. Preoperative or normal tracheal segments adjacent to the stenotic area were used to obtain normal tracheal diameter measurements. Planimetrically determined cross-sectional area (CSA), obtained from pre- and postoperative tracheoscopic photographs, was used to calculate PTLS. The CSA of tracheal specimens obtained at necropsy was determined, using the formula for an ellipse. Percentage of luminal stenosis was calculated, using CSA of the stenotic site and of segments craniad and caudad to the site obtained at necropsy or at surgery. All methods were compared with the control method of planimetrically determined CSA of sections obtained at necropsy of the tracheostomy and segments craniad and caudad to the site. RESULTS: Correlation was poor for radiographic and tracheoscopic techniques (r = 0.146 to 0.458, P > 0.05) The formula for an ellipse accurately predicted PTLS when measurements obtained at surgery (r = 0.516, P = 0.049) or segments craniad and caudad (r = 0.853, P < 0.001) to the site were used. CONCLUSION: Antemortem methods of assessing PTLS did not correlate with control planimetric methods. Methods using CSA determined by tracheal diameter were weakly correlated to control planimetric techniques. CLINICAL RELEVANCE: Accurate measurement of the degree of tracheal stenosis cannot be made in clinical patients using current techniques. PMID- 9328653 TI - Measurement of cyclic variation in ultrasonic integrated backscatter in conscious, unsedated, clinically normal dogs. AB - OBJECTIVE: To assess the feasibility and repeatability of measuring ultrasonic integrated backscatter in unsedated conscious dogs, using a protocol previously validated in pigs with open thorax. ANIMALS: 11 clinically normal conscious unsedated German Shorthair Pointers. PROCEDURE: A modified commercially available echocardiography system was used to record long-axis views of the heart. The radiofrequency data from 15 consecutive frames were digitized and analyzed. Regions of interest were chosen within the myocardium, and the ultrasonic integrated backscatter within each region was calculated in the time domain for each frame. RESULTS: Cyclic variation in integrated backscatter values was observed, with maximal values at end-diastole and minimal values at end-systole. Mean +/-SD amplitude of cyclic variation was 5.81 +/- 3.86 dB over all the regions chosen. CONCLUSIONS: Results agreed with those obtained by other investigators working with dogs with open thorax and those with closed thorax while under general anesthesia. The analysis of the components of variance indicates that this is a consistent, reliable technique in conscious unsedated dogs. CLINICAL RELEVANCE: Integrated ultrasonic backscatter measurement provides a noninvasive means of tissue characterization. Use of this protocol reliably yields cyclic variation in integrated backscatter and could be applied clinically to dogs with myocardial disease. PMID- 9328654 TI - Measurement of total body water content in horses, using deuterium oxide dilution. AB - OBJECTIVE: To measure total body water (TBW) content in horses, using deuterium oxide (D2O) dilution. ANIMALS: Six 8- to 10-year-old healthy untrained mixed breed horses, weighing (mean +/-SD) 503.4 +/- 64.0 kg. PROCEDURE: After a 12-hour nonfeeding period, 6 horses were given D2O (0.14 g/kg of body weight) via nasogastric tube. Blood samples were collected from a preplaced indwelling jugular vein catheter prior to and 1 to 8, 10, 12, 14, and 24 hours after administration of D2O. Blood samples were centrifuged immediately, and plasma was collected and stored at -70 C until analysis. The D2O content in plasma was measured by zinc reduction to deuterium gas. The resulting gas was measured, using an isotope ratio mass spectrometer. RESULTS: Deuterium oxide was rapidly absorbed from the gastrointestinal tract of all horses, and reached peak (mean +/ SD) plasma concentration (1,454.4 +/- 163 delta D/ml or parts/thousand) 1 hour after administration. Plasma concentration decreased slowly during the next 2 to 3 hours, then remained statistically constant from 2 to 5 hours (early plateau phase) and 3 to 7 hours (late plateau phase) after administration. Mean +/- SEM TBW content was 623.0 +/- 2.2 ml/kg (62.3% of body weight) for the early plateau phase and 630.3 +/- 2.2 ml/kg (63.0% of body weight) for the late plateau phase. CONCLUSION: Deuterium oxide dilution appears to be of value for measurement of TBW content in horses, and has a 4-hour plateau effect. Equilibration of D2O with large intestinal water may be the reason for the prolonged equilibrium time and plateau effect seen in these horses. CLINICAL RELEVANCE: Deuterium oxide appears safe and efficacious for determining TBW content in horses and may be helpful for determining changes in TBW content during exercise and disease. PMID- 9328655 TI - Evaluation of noninvasive monitoring techniques in domestic ferrets (Mustela putorius furo). AB - OBJECTIVE: To evaluate instrument placement and accuracy of indirect physiologic monitoring techniques in anesthetized domestic ferrets. ANIMALS: 10 healthy adult female ferrets (Mustela putorius furo). PROCEDURE: Direct arterial blood pressure measurement and arterial blood sample collection were performed in ferrets. A pulse oximeter probe was clipped to a forefoot or hind foot; an airway adaptor for capnography was attached to the endotracheal tube; and a sphygmomanometer cuff and Doppler flow probe were positioned on the tail. Isoflurane and nitrous oxide concentrations were varied to induce episodes of hypotension or hypoxia, respectively. Aforementioned noninvasive techniques were compared with direct methods of arterial blood gas analysis, hemoximetry, and arterial blood pressure measurement. Simultaneously obtained direct and indirect measurements were statistically evaluated for mean and SD of the differences, and SEM, and subjectively, for ease of use and relevance to the clinical situation. RESULTS: Values obtained from pulse oximetry were closely related to oxygen saturation measured by blood gas analysis (O2sat). The mean (+/- SD) difference for all results was -0.49 (+/- -4.09)%. The most precise measurements were obtained when O2sat was between 90 and 100%. Capnography measurements varied between ranges. The most accurate measurements were obtained when PaCO2 was < 25 mm of Hg, when the mean difference was 1.6 (+/- -3.01) mm of Hg. Indirect blood pressure measurement consistently underestimated the direct blood pressure value. CONCLUSIONS AND CLINICAL RELEVANCE: Pulse oximetry is a convenient and accurate method for monitoring oxygen saturation in domestic ferrets. Capnography is useful for monitoring respiratory rate and pattern, but may present difficulties in interpretation of actual PaCO2. Indirect blood pressure monitoring is not accurate by use of current methods in ferrets. PMID- 9328657 TI - Within- and between-examiner repeatability of distraction indices of the hip joints in dogs. AB - OBJECTIVE: To evaluate in vivo repeatability of the distraction index method of evaluating hip joint laxity in dogs. ANIMALS: 31 two-year-old Labrador Retrievers. PROCEDURE: Each dog was anesthetized and radiographically evaluated for hip joint laxity 4 times: twice by an experienced examiner and twice by an examiner who had no previous knowledge of or training in the technique prior to the first day of testing. Distraction indices (DI) were determined from the radiographs and intraclass correlation coefficients were calculated to evaluate the repeatability of DI measurements between and within examiners. RESULTS: Intraclass correlation coefficients were high (range, 0.85 to 0.94). Lower limits of the 95% confidence intervals for the intraclass correlation coefficients ranged from 0.75 to 0.89. CONCLUSIONS: Between- and within-examiner repeatabilities of DI measurements were high, suggesting that the technique is clinically reliable. CLINICAL RELEVANCE: Distraction index is a reliable measure of hip joint laxity and a good predictor of the risk of development of degenerative joint disease associated with hip dysplasia in dogs. Establishment of high repeatability of DI measurements suggests that the stress-radiographic method may be used by multiple examiners with the expectation of comparable and consistent results. PMID- 9328656 TI - Application of multiplex polymerase chain reaction for rapid identification of Campylobacter jejuni and C coli associated with reproductive failure. AB - OBJECTIVE: To evaluate a multiplex polymerase chain reaction (PCR) to distinguish Campylobacter jejuni from C coli as causes of reproductive failure. PROCEDURE: Review of clinical cases of reproductive failure attributed to C jejuni or C coli. RESULTS: A case of swine abortion was attributable to infection with C coli. The porcine abortion isolates were verified as C coli by restriction fragment length polymorphism and multiplex PCR. Cases of endometritis in a fox and in mink caused by C jejuni were reviewed, and isolates were confirmed as C jejuni by results of the multiplex PCR. CONCLUSION: Multiplex PCR was useful in identifying C coli and C jejuni recovered from atypical cases of reproductive failure. Multiplex PCR in conjunction with conventional assays may be useful for verifying other unusual instances of campylobacteriosis. PMID- 9328659 TI - Developmental variation in lumbosacropelvic anatomy of thoroughbred racehorses. AB - OBJECTIVE: To describe the incidence and types of gross osseous developmental variations and ages of physeal closure in the caudal portion of the thoracic and lumbosacral spine and the pelvis in a sample of Thoroughbred racehorses. ANIMALS: Thoroughbred racehorses (n = 36) that died or were euthanatized at California racetracks between October 1993 and July 1994. PROCEDURE: Lumbosacropelvic specimens were collected, and all soft tissues were removed. The osseous specimens were visually examined. RESULTS: Only 22 (61%) specimens had the expected number of 6 lumbar and 5 sacral vertebrae. Eight (22%) specimens had thoracolumbar transitional vertebrae, and 13 (36%) had sacrocaudal transitional vertebrae. Articular process asymmetries were present at 1 or more vertebral segments in 30 (83%) specimens. Intertransverse joints (2 to 4 pairs/specimen) were bilaterally distributed in the caudal portion of the lumbar spine and the lumbosacral joint in 31 (86%) specimens. Five (14%) specimens had asymmetric distribution of the intertransverse joints. Intertransverse joint ankylosis was found in 10 (28%) specimens. Lumbosacral vertebral body physeal closure occurred between 4.9 and 6.7 years of age; pelvic physeal closure occurred between 5.2 and 5.8 years of age. Iliac crest and ischial arch epiphyseal formation was evaluated, using a grading system, and fusion to the underlying bone occurred at 7.2 years and 5.4 years of age, respectively. CONCLUSIONS: Numerous vertebral anatomic variations were commonly found in a sample of Thoroughbred racehorses. CLINICAL RELEVANCE: Normal anatomic variations and ages of skeletal maturity need to be considered in clinical evaluation of the equine spine and pelvis for differentiation from pathologic findings. PMID- 9328658 TI - Biomechanical characterization of passive laxity of the hip joint in dogs. AB - OBJECTIVE: To investigate the in vitro load/displacement characteristics of the hip joints in dogs as a function of joint position. SAMPLE POPULATION: 10 hip joints from 5 healthy dogs. PROCEDURE: A material test system was used to generate load/displacement curves for each joint. Joints were mounted in a custom designed jig that held the joint in fixed anatomic orientations while plotting displacement and corresponding applied loads. All hips were cycled between 40 N of compression and 80 N of distraction. Each hip was tested at 10 degrees increments from 30 degrees flexion to 70 degrees extension. RESULTS: When the hips were in a neutral orientation (approximately a standing position), load/displacement curves were characteristically sigmoidal (tri-phasic), indicating that, in this position, displacement was not highly dependent on load. The curves had a central low-stiffness region in which most of the lateral displacement took place. In contrast, when hips were positioned at the extremes of flexion and extension, this central, low-stiffness region was less distinct, and load/displacement curves were more linear, indicating a proportional relation between load and displacement. The load/displacement curve of 1 hip joint in the study deviated markedly from the others in a pattern consistent with cavitation of the synovial fluid. CONCLUSIONS: When the hip joint is positioned in a neutral position, load-displacement behavior is sigmoidal, whereas when the hip joint is in an extended position, load/displacement behavior is more linear. CLINICAL RELEVANCE: Establishing load/displacement behavior of the hip joints in dogs was an important exercise in establishing the position for and estimating the repeatability of a clinical stress-radiographic method for quantitating joint laxity in dogs. PMID- 9328660 TI - Flow cytometric method for detecting thiazole orange-positive (reticulated) platelets in thrombocytopenic horses. AB - OBJECTIVE: To evaluate a method for detecting thiazole orange-positive (TO+, reticulated) platelets in equine blood, using flow cytometry. ANIMALS: 16 healthy, equine infectious anemia virus (EIAV)-negative horses and ponies; 9 thrombocytopenic, EIAV-positive horses and ponies; and 2 thrombocytopenic, EIAV negative horses. PROCEDURE: Blood from healthy and thrombocytopenic horses was collected by jugular venipuncture. Appropriate sample requirement and incubation time for the assay were evaluated, using blood anticoagulated with EDTA or sodium citrate, or platelet-rich plasma in sodium citrate. The sample of blood or platelet-rich plasma was incubated with thiazole orange, and flow cytometric analysis was performed. Percentage of circulating TO+ platelets was determined from fluorescence (FL-1) logarithmic histograms. RESULTS: Healthy ponies (n = 9) had 1.28 to 2.83% (mean +/- SD, 2.03 +/- 0.50%) and horses (n = 7) had 0.9 to 3.44% (2.12 +/- 1.14%) TO+ platelets in circulation. Thrombocytopenic ponies (n = 7) had 11.14 to 48.41% (26.51 +/- 11.99%) and thrombocytopenic horses (n = 4) had 2.33 to 8.52% (6.19 +/- 2.68%) TO+ platelets in circulation. Mean platelet counts for the thrombocytopenic ponies and horses were 24,400 +/- 20,500 and 39,300 +/- 13,500 platelets/microliters, respectively (reference range, 94,000 to 232,000 platelets/ microliters). CONCLUSION: Thiazole orange-positive platelets can be detected in equine blood and percentages of TO+ platelets are increased in thrombocytopenic horses. CLINICAL RELEVANCE: Enumeration of TO+ platelets may prove to be a helpful noninvasive clinical measurement of bone marrow platelet production and aid in the assessment of platelet kinetics in thrombocytopenic horses. PMID- 9328661 TI - Risk factors associated with the likelihood of leptospiral seropositivity in horses in the state of New York. AB - OBJECTIVE: To determine and quantify risk factors associated with exposure of horses to the following serovars of Leptospira interrogans: pomona, autumnalis, and bratislava. ANIMALS: 2,551 horses were randomly selected from a target population during the period of May 1991 to August 1993. PROCEDURES: Blood was collected from the horses and tested for antibodies to serovars, using the microscopic agglutination test. A titer > or = 1:100 indicated seropositivity. Information was collected on each horse, its environment, and each farm's management practices. Logistic regression analysis was used to develop a multidimensional indexing system for indices of exposure and to identify factors significantly-associated with the risk of seropositivity. These indices were: 1) rodent exposure; 2) wildlife exposure; 3) soil and water; and 4) management. RESULTS: Rodent exposure index value was associated with the risk of exposure to all 3 serovars. Management index value was positively associated with the risk of exposure to serovars pomona and bratislava, but not with risk of exposure to serovar autumnalis. Soil and water index value had a positive association with risk of exposure to serovars pomona and autumnalis, but not to serovar bratislava. The wild-life index value and the population density of horses turned out together were associated with the risk of exposure to serovar autumnalis. Age of horse in years was associated nonlinearly (years) and linearly (years) with the risk of exposure to serovars autumnalis and bratislava, and only linearly with the risk of exposure to serovar pomona. CONCLUSION: Risk of seropositivity to the 3 serovars of L interrogans varies according to age, management practices, population density of horses turned out together, and the values of the rodent exposure, wildlife exposure, and soil and water indices. PMID- 9328662 TI - Safety and efficacy of a mutagen-attenuated Rift Valley fever virus vaccine in cattle. AB - OBJECTIVE: To examine safety and efficacy of a mutagen attenuated Rift Valley fever virus (RVFV) vaccine (RVF MP-12) in cattle. ANIMALS: 38 pregnant cows, 14 steers, and 10 lactating dairy cows. PROCEDURE: Pregnant cows in their third, fifth, or eighth month of gestation were vaccinated (1 ml of RVF MP-12 containing 5 log10 plaque-forming units [PFU] of virus) and were monitored daily through parturition for signs of disease, viremia, and immunologic response. Additionally, 10 vaccinated pregnant cows were challenge inoculated with virulent RVFV at post-vaccination day (PVD) 30 and were monitored daily for untoward effects. Ten unvaccinated pregnant cows also were challenge inoculated with virulent RVFV and served as challenge controls. Vaccinated lactating dairy cows were monitored for viremia and virus shedding in the milk through PVD 14. Yearling steers were vaccinated to assess their immunologic response to various doses of vaccine and were challenge inoculated with virulent RVFV at PVD 28 to assess protection. RESULTS: 10 of 38 (26.3%) cows vaccinated during pregnancy developed transient postvaccination viremia titer > or = 2.5 log10 PFU/ml of serum. All vaccinated cows delivered live, healthy calves that were RVFV seronegative at birth, but which quickly acquired colostral antibodies. Vaccinated cows and their fetuses were protected when challenge exposed with virulent RVFV at PVD 30, whereas unvaccinated pregnant cows inoculated with RVFV became febrile and viremic, and aborted. Vaccine virus was unsuccessfully sought from milk of lactating dairy cows after vaccination, suggesting that shedding of vaccine virus through milk should not be a concern. Steers, inoculated with tenfold escalating vaccine doses, beginning with 1.0 log10 PFU, were protected against virulent RVFV challenge exposure. CONCLUSIONS: RVF MP-12 may be safe and efficacious for use in pregnant or lactating bovids, and a minimal dose of vaccine may provide suitable protection against viremia. PMID- 9328663 TI - Safety of a mutagen-attenuated Rift Valley fever virus vaccine in fetal and neonatal bovids. AB - OBJECTIVE: To examine effects of in utero inoculation with a mutagen-attenuated Rift Valley fever virus (RVFV) vaccine (RVF MP-12) on fetal bovids and to assess the safety and efficacy of calfhood vaccination with RVF MP-12. ANIMALS: 18 pregnant Hereford and Hereford-type cows in the third or fifth month of gestation, their progeny, and 25 calves from cows immunized with RVF MP-12 during pregnancy. PROCEDURE: Bovine fetuses were inoculated, via laparotomy, with 1 ml of RVF MP-12 containing 5 log10 plaque-forming units (PFU) of virus. Blood was obtained from newborn calves prior to their ingestion of colostrum. Immune-naive calves and calves born to RVF MP-12-vaccinated dams, ranging in age from 2 to 45 days, were vaccinated with RVF MP-12, and some were later challenge exposed with 1 ml of 5.7 log10 PFU of virulent RVFV strain ZH-501. Cows were monitored for viremia and antibody responses and for hematologic and serum biochemical alterations through parturition or abortion. RESULTS: Surviving in utero vaccinated calves were healthy, with no noticeable defects. Except for 1 vaccine inoculated fetus that died on postinoculation day 21, all in utero-vaccinated fetuses had serum neutralizing antibody titer > or = 1:20 at the time of delivery. All dams of in utero-vaccinated fetuses also developed neutralizing antibody titer. Calves born to cows vaccinated during gestation did not have antibody at birth, and all but 1 quickly acquired colostral antibody. Postparturient inoculation of immune-naive calves and calves with colostral antibodies resulted in no untoward effects, and all calves with detectable neutralizing antibodies were protected against virulent virus challenge exposure. CONCLUSIONS: Fetal death and abortion would be rare even if fetuses were exposed to RVF MP-12. The trauma and complications associated with in utero inoculation do not make this a practical method of immunization. RVF MP-12 was safe, immunogenic, and protective in calves as young as 2 days of age. PMID- 9328664 TI - Comparison of the PK(15)- and WEHI 164 (clone 13)-based bioassays for detection of porcine tumor necrosis factor. AB - OBJECTIVE: To determine relative sensitivities of the PK(15)- and WEHI 164(13) based bioassays for detection of tumor necrosis factor alpha (TNF). SAMPLE POPULATION: Recombinant human, murine, and porcine INF, and serum from pigs given endotoxin IV. PROCEDURE: Two cell lines were used as targets for recombinant human, murine, and porcine TNF cytotoxicity bioassays. Pigs were given sublethal doses of endotoxin to obtain serum samples containing high activity of porcine TNF. Serum TNF activity was tested, using both cell lines. Viable cells were detected by addition of dimethylthiazol diphenyltetrazolium bromide after 18 to 20 hours' incubation with samples containing TNF. RESULTS: The 2 cell lines tested had different sensitivities to human, murine, and porcine TNF. Compared with WEHI 164(13) cells, PK(15) cells were 50 times less sensitive to murine TNF and 15 times less sensitive to human TNF. However, PK(15) cells were 4 times more sensitive to recombinant porcine TNF and 15 times more sensitive to porcine serum containing TNF. CONCLUSIONS: The PK(15) cell line was more sensitive to porcine TNF-mediated lysis than was the WEHI 164(13) cell line. The PK(15)-based TNF bioassay will be especially useful for study of infectious disease processes in swine, particularly where low activity of TNF exists. PMID- 9328666 TI - Induction of gross and microscopic lesions of porcine proliferative enteritis by Lawsonia intracellularis. AB - OBJECTIVE: To evaluate experimental induction of porcine proliferative enteritis (PPE), using cell cultured Lawsonia intracellularis (ileal symbiont intracellularis), and to determine whether dexamethasone administration or age of the host or both affects susceptibility to L intracellularis infection. ANIMALS: Thirty-two 3- or 7-week-old pigs. PROCEDURES: Lawsonia intracellularis was extracted from tissue with lesions of PPE and was subcultured in a continuous Henle 407 cell line at 37 C under an atmosphere of 5% CO2. Three- or 7-week-old pigs were inoculated orally with 100 ml of a 10-day-old cell culture preparation of the bacterium or infective intestinal homogenates. Control pigs were inoculated with uninfected Henle cells. Pigs were observed daily for clinical signs of infection and necropsied at death or at termination of the study. Lesions in the small and large intestines were recorded. RESULTS: Diarrhea was observed in pigs 4 to 7 days after inoculation with the pure culture agent or homogenates and lasted throughout the study period. Histologic lesions consistent with PPE were detected in pigs inoculated with pure culture. Intestinal lesions were absent in control pigs inoculated with uninfected Henle cells. Differences in lesions were not significant between treatment groups that varied in age or were receiving dexamethasone. Tissue specimens from pigs at necropsy were culture negative for Salmonella spp and Serpulina hyodysenteriae. CONCLUSION AND CLINICAL RELEVANCE: Gross and microscopic lesions typical of acute PPE were induced in pigs by use of a cell culture agent. Age differences and the stress induced by administration of dexamethasone had no effect on development of intestinal lesions. PMID- 9328665 TI - Infection of polarized epithelial cells with enteric and respiratory tract bovine coronaviruses and release of virus progeny. AB - OBJECTIVE: To investigate the susceptibility of polarized epithelioid human rectal tumor (HRT-18G) cells to bovine coronaviruses (BCV) isolated from enteric (EBCV) and respiratory (RBCV) tract infections. PROCEDURE: Cells of the G clone of HRT-18 were grown to confluent monolayers on permeable supports, and were directionally infected at the apical and basolateral domains with 3 wild-type BCV strains, RBCV-LSU-94LSS-051-2, RBCV-OK-0514-3, and EBCV-LY138-2, and 1 cell culture-adapted strain, EBCV-L9-80. Sequential cytopathic changes were microscopically monitored. Medium samples for titration of hemagglutinins and viral infectivity were collected directionally from both domains of the infected cell cultures at various intervals. RESULTS: Polarized epithelioid HRT-18G cells from apical domains had maximal susceptibility to infection with the EBCV and RBCV strains, and those from basolateral surfaces had minimal susceptibility. Titers of hemagglutinins and infective progeny BCV reached 1,280 hemagglutinin units and 4.2 x 10(8) plaque-forming units/ml for apical samples, but were minimal for basolateral samples. Asymmetric virus release occurred through the apical surfaces of the HRT-18G cells by 12 hours after infection when cell fusion as a sign of cytopathic changes began. When cells were infected basolaterally, progeny virions released from apical surfaces reinfected the target cells from the apical domains and induced cytopathic changes were delayed about 12 hours, compared with changes detectable in apically exposed cultures. CONCLUSIONS: EBCV and RBCV, isolated from cattle, had marked tropism for polarized epithelioid HRT 18G cells. Entry of BCV into the polarized HRT-18G cells was effected maximally through the apical domains and minimally through the basolateral domains. Release of progeny BCV occurred preferentially from the apical domains. PMID- 9328667 TI - Effects of intravenous administration of sodium hyaluronate on carpal joints in exercising horses after arthroscopic surgery and osteochondral fragmentation. AB - OBJECTIVE: To evaluate the effects of arthroscopic surgery, osteochondral fragmentation, and treatment with IV administered hyaluronate on histologic, histochemical, and biochemical measurements within the carpal joints of horses. ANIMALS: 12 clinically normal horses, 2 to 7 years of age. PROCEDURE: Horses had an osteochondral fragment created at the distal aspect of the radiocarpal bone of 1 randomly chosen middle carpal joint to simulate osteochondral fragmentation. Horses were treated with 40 mg of hyaluronate or saline solution (placebo) intravenously once a week for 3 consecutive weeks (days 13, 20, and 27 after surgery). Treadmill exercise proceeded 5 days per week beginning 15 days, and ending 72 days, after surgery. Clinical evaluations were performed at the beginning and end of the study. Synovial fluid samples were obtained aseptically from both middle carpal joints on days 0, 13, 20, 27, 34, and 72 after surgery, and total protein, inflammatory cell, hyaluronate, glycosaminoglycan, and prostaglandin E2 concentrations were measured in each sample. All horses were euthanatized on day 72. Synovial membrane and articular cartilage were obtained for histologic evaluation. Articular cartilage samples were also obtained aseptically for determining glycosaminoglycan content and chondrocyte synthetic rate for glycosaminoglycans. RESULTS: Horses treated with hyaluronate intravenously had lower lameness scores (were less lame), significantly better synovial membrane histologic scores, and significantly lower concentrations of total protein and prostaglandin E2 within synovial fluid 72 days after surgery, compared with placebo-treated horses. Treatment with intravenously administered hyaluronate had no significant effects on glycosaminoglycan content, synthetic rate or morphologic scoring in articular cartilage, or other synovial fluid measurements. CONCLUSION: Intravenously administered hyaluronate appears to alleviate signs of lameness by interacting with synoviocytes, and by decreasing production and release of inflammatory mediators. PMID- 9328668 TI - Effects of valacyclovir in cats infected with feline herpesvirus 1. AB - OBJECTIVE: To determine whether orally administered valacyclovir can be used safely and effectively to treat cats with primary, feline herpesvirus 1 (FHV-1) infection. ANIMALS: 14 specific-pathogen-free adult cats. PROCEDURE: Cats were infected with FHV-1 strain 87-727 (300 microliters, 10(7) plaque-forming units/ml) by ocular and nasal inoculations, and were treated every 6 hours with dextrose (controls) or valacyclovir (60 mg/kg of body weight, PO). Virus shedding from both eyes and the oropharynx was monitored every 2 days by virus isolation, and subjective clinical scores were assigned daily for ocular and nasal discharge and conjunctival hyperemia. Urinalysis, CBC, and serum biochemical analysis were done prior to inoculation, and on days 2, 5, 7, 9, and 12 of infection. Differences in CBC and serum biochemical indices between groups were compared, as were differences between preinfection values and maximal postinfection values, rectal temperature, and scores for disease severity. RESULTS: All cats developed acute conjunctivitis and rhinitis typical of FHV-1 infection. Beginning between days 6 and 9, valacyclovir-treated cats became noticeably more lethargic and dehydrated than did cats of the control group. Total WBC and neutrophil counts were significantly lower in cats of the valacyclovir group. The experiment was terminated on day 12 for humane reasons. Histologic changes attributable to FHV-1 infection were similar in all cats. Additional histologic abnormalities seen only in the valacyclovir-treated cats were coagulative necrosis of the renal tubular epithelium, centrilobular atrophy and hepatic necrosis, and severe bone marrow depression. CONCLUSIONS: Cats appear to be uniquely sensitive to the toxic effects of valacyclovir, and even high doses appear not to suppress FHV-1 replication in acutely infected cats. CLINICAL RELEVANCE: Use of valacyclovir is of questionable value in cats with acute FHV-1 infection and, at high doses, the drug may be toxic. PMID- 9328669 TI - Role of endothelium and nitric oxide in the in vitro response of equine colonic venous rings to vasoconstrictor agents. AB - OBJECTIVE: To determine in vitro contractile responses of equine colonic veins to various vasoconstrictor agents. ANIMALS: Colonic veins collected from 8 adult horses. PROCEDURE: Veins were cut into 4-mm-wide rings, placed in organ baths at 37 C, and attached to a force-transducer interfaced with a polygraph; 2 g of tension was applied, and rings were allowed to equilibrate for 45 minutes. Bath solution was replaced, and tension was reapplied at 15-minute intervals. Cumulative concentration responses (10(-8) to 10(-4) M) were determined for each agent, using separate rings (n = 8). Three vein groups were evaluated: endothelium-intact, endothelium-denuded, and N omega-nitro-L-arginine methyl ester (L-NAME, 10(-5) M)-treated. Maximal responses by each vein to each agent were considered 100%; responses to lower concentrations were calculated as percentage of maximum. RESULTS: Considering all vein groups, comparison of the doses that caused 50% of the maximal contraction revealed relative sensitivity of colonic veins to be: angiotensin II (ANG) > thromboxane B2 analogue (TXB) > 5 hydoxytryptamine (5HT) > norepinephrine (NE) > histamine (HST) > prostaglandin F2 alpha (PGF) > vasopressin (VP). Compared with ANG, PGF, TXB, and VP, treatment with HST, 5HT, and NE evoked significantly greater responses. Endothelium-denuded and L-NAME-treated colonic veins had significantly greater maximal contractile responses than did endothelium-intact veins. CONCLUSIONS: Response of colonic veins to vasoconstrictor agents was differential; sensitivity was not altered by endothelium removal or L-NAME treatment; maximal responses of endothelium-intact veins were greater than those of endothelium-denuded and L-NAME-treated veins; and responses of endothelium-denuded and L-NAME-treated veins were not different. CLINICAL RELEVANCE: Alterations in colonic veins that mimic conditions associated with large-colon volvulus may contribute to blood flow alterations, edema formation, and vascular responses to hypovolemic and endotoxic shock. PMID- 9328670 TI - Isolation and expression of a porcine lactoferrin gene. AB - OBJECTIVE: To elucidate the spatial and temporal expression of a porcine lactoferrin (LTF) gene. ANIMALS: 4 female and 4 male Large White pigs. PROCEDURES: We examined LTF expression in various organs excised from the pigs, using northern blot hybridization with a porcine LTF cDNA probe. Antibodies against porcine LTF were raised in rabbits and were used along with immunohistochemical staining to localize the LTF protein. RESULTS: High amounts of porcine LTF mRNA were detected in the secreting mammary gland and epididymis. This distribution is consistent with that of porcine LTF examined by immunohistochemistry. In female pigs, porcine LTF mRNA concentration increased remarkably in the ductal cells of the lactating mammary gland then significantly decreased at day 21 after parturition. Furthermore, specific staining for LTF was observed in the epithelial cells of the gastrointestinal tract of female pigs, but not in the uterus, ovaries, spleen, kidneys, pancreas, muscles, heart, brain, lungs, or liver of postpartum female pigs, or in the testes of male pigs. CONCLUSIONS: Gene expression of porcine LTF is closely related to lactation in the mammary gland. Distribution of LTF in the epididymis suggests that LTF may have a regulatory role in development of the reproductive tract of male pigs. PMID- 9328671 TI - Mechanisms of acute intraocular pressure increases after phacoemulsification lens extraction in dogs. AB - OBJECTIVE: To investigate the mechanisms by which intraocular pressure (IOP) increases acutely after phacoemulsification (PE) lens extraction in clinically normal dogs. ANIMALS: 24 young adult dogs. PROCEDURE: Intraocular pressure was monitored for up to 24 hours after unilateral intercapsular PE in 17 clinically normal, adult dogs. In 8 of these dogs, use of 2% hydroxypropyl methylcellulose (HPMC) aided capsulorhexis. Mean volume of irrigation, PE time, and power were constant between groups. After surgery, dogs were randomized then euthanatized, and eyes were examined grossly and histologically at 0 (n = 4), 3 (n = 7), and 24 hours (n = 6) after PE. Seven additional dogs underwent anterior chamber decompression alone (n = 4) or served as morphologic controls (n = 3). RESULTS: Intraocular pressure peaked by postoperative hour 3 at 49.9 +/- 5.0 mm of Hg and normalized by 24 hours. Use of HPMC did not affect the peak or duration of IOP increase. Blood refluxed into the collecting channels and corneoscleral trabecular meshwork in operated eyes. Computer-aided morphologic analysis indicated significant (P < 0.001) reduction in ciliary cleft cross-sectional surface area and width immediately after PE, but not after anterior chamber decompression alone. Cleft collapse was significantly (P < 0.02) greater at 24 than at 3 hours, despite return of IOP to control values by 24 hours. Plasmoid aqueous also was found in the meshwork. CONCLUSIONS AND CLINICAL RELEVANCE: Sudden, large increases in IOP with few overt clinical signs may occur immediately after lens extraction in dogs. Such increases risk compromising the corneal incision and may damage the optic nerve, thereby complicating lens extraction. Structural alterations in the trabecular meshwork persist after IOP has normalized in 24 hours and may contribute to genesis of glaucoma in the late postoperative period. PMID- 9328672 TI - Effects of postoperative peritoneal lavage on pharmacokinetics of gentamicin in horses after celiotomy. AB - OBJECTIVE: To evaluate the effect of peritoneal lavage on pharmacokinetics of gentamicin sulfate in healthy horses after experimental celiotomy. ANIMALS: 13 clinically normal horses. PROCEDURE: Horses were randomly assigned to control or experimental groups. All horses received gentamicin (6.6 mg/kg of body weight, IV, q 24 h) before surgery, underwent experimental abdominal surgery, and had abdominal drains placed percutaneously. Horses of the experimental group received postoperative peritoneal lavage; horses of the control group did not receive peritoneal lavage. The day after surgery, 24 hours after the preoperative dose of gentamicin, a second dose of gentamicin was administered. Three and 15 hours after this second dose of gentamicin, horses of the experimental group received peritoneal lavage. Venous blood was obtained, for determination of concentration of gentamicin, immediately before and at specified intervals during the 24-hour period after the second dose of gentamicin. RESULTS: There were no differences in any of the pharmacokinetic values of gentamicin between horses of the control and experimental groups. CONCLUSIONS: Peritoneal lavage had no effect on pharmacokinetics of gentamicin in healthy horses after abdominal surgery, in which localized nonseptic peritonitis was induced. CLINICAL RELEVANCE: Peritoneal lavage in horses with localized nonseptic peritonitis or for the prevention of intra-abdominal adhesions should not necessitate alteration of the dosage of gentamicin to maintain predictable serum concentrations. PMID- 9328673 TI - Effects of multiple freezing-thawing cycles on ultimate indentation load and stiffness of bovine cancellous bone. AB - OBJECTIVE: To assess the effects of multiple freezing-thawing cycles on ultimate indentation load and stiffness of bovine tibial cancellous bone with regard to freezing conditions (saline solution or air) or methods of thawing (saline solution or air). SAMPLES: 4 tibias from 4 adult cows. PROCEDURE: The proximal portions of the tibias were sectioned to produce 20-mm-thick bone slices. Slices were subjected to 5 freezing-thawing cycles under 4 conditions: freezing with and without saline solution, then thawing in saline solution or exposed to the air. The mechanical properties of the bone slices before and after the treatments were measured, using an indentation test for comparison. Indentation testing was performed before and after freezing-thawing cycles to measure differences in mechanical parameters. RESULTS: Significant differences in mechanical parameters were not found for bone specimens frozen in saline solution and thawed in saline solution; frozen without saline solution and thawed in air; frozen in saline solution and thawed in air; or frozen without saline solution and thawed in saline solution. CONCLUSIONS: Multiple thawing-freezing cycles do not significantly affect the ultimate indentation load and stiffness of bovine tibial cancellous bone. PMID- 9328674 TI - Fluoride release of restorative materials exposed to a fluoridated dentifrice. AB - The purpose of this study was to examine the effect that brushing with a fluoridated dentifrice (Crest--Procter and Gamble) has on the fluoride release of restorative materials. Thirty standardized discs were fabricated; 10 were P-50 (3M) nonfluoridated composite resin (control), ten were Heliomolar Radiopaque (Ivoclar/Vivadent) fluoride releasing composite resin, and ten were Ketac Fil (ESPE) glass ionomer cement. Specimens were placed into separate containers of 10 mL deionized water. Half the specimens from each group were brushed with fluoridated dentifrice for two minutes twice per day and rinsed. The fluoride level of each specimen was evaluated for thirty days, using a fluoride specific ion analyzer. An analysis of variance (ANOVA) and Duncan's test (p < 0.05) indicated significant differences in fluoride release. The brushed glass ionomer was significantly higher than all other groups and the glass ionomer not brushed was significantly higher than all composite groups. Glass Ionomer Cement-brushed > Glass Ionomer Cement > Fluoridated Composite Resin-brushed = Fluoridated Composite Resin = Composite Resin-brushed = Composite Resin. Brushed glass ionomer appears to release the highest fluoride level, acting as a fluoride reservoir from the dentifrice for subsequent fluoride release. PMID- 9328675 TI - Calcium hydroxide pulpotomy with a light-cured cavity-sealing material after two years. AB - In this follow-up study a light-cured glass ionomer lining cement was evaluated as a cavity-sealing material in calcium hydroxide pulpotomies in primary molars after one and two years. The pulpotomy dressing was a suspension of pure calcium hydroxide with either tap water or sterile saline. The success rate of the pulpotomies after one year was 87.7 percent and after two years 80.4 percent (clinically and radiographically). This result was only influenced by the type of restoration (amalgam versus stainless steel crown). The results of the present investigation compare favorably with those of other published studies of pulpotomy of primary molars using calcium hydroxide as the wound dressing. PMID- 9328676 TI - Pulp response to ferric sulfate, diluted formocresol and IRM in pulpotomized primary baboon teeth. AB - This study investigated the pulp response to a 15.5 percent ferric sulfate solution (FS) and a 20 percent dilution of formocresol (DFC) in pulpotomized primary teeth of baboons, after four and eight weeks. Pulpotomies were performed in seventy-nine primary teeth of 4 baboons. After coronal pulp resection, the pulp stumps were painted with ferric sulfate for fifteen seconds, in thirty-two teeth (group 1); in another thirty-two teeth, a cotton pellet moistened with dilution of formocresol was placed over the pulp stumps for five minutes, and removed (group 2). In fifteen teeth, IRM was placed directly over the pulp stumps after hemostasis (group 3--control). The teeth of all groups were sealed with IRM, and examined for inflammatory changes under a microscope by two blinded examiners. Seventy-seven teeth were assessed. Mild or no inflammation was found in 58 percent (18/31) of the teeth of group 1, in 48 percent (15/31) of those of group 2, and in 73 percent (11/15) of those of group 3. Severe inflammation was found in 35 percent (11/31) of group 1, 29 percent (9/31) of group 2, and in 7 percent (1/15) of group 3. No statistically significant difference between the three groups was observed for degree of inflammation, periradicular or interradicular abscess or inflammatory root resorption (chi-square p > 0.05). Dentin bridges were observed in 52 percent (16/31) of the teeth in group 1, 52 percent (16/31) of those of group 2, and in 73 percent (11/15) of those of group 3. No difference was found between the experimental and control groups for the presence of dentin bridge, (p > 0.05). Ferric sulfate produced pulp responses that compared favorably to those of diluted formocresol. PMID- 9328678 TI - Consequences of serious oral injury associated with the congenital analgia syndrome. AB - Three sisters at the ages of seven months, twelve years, and thirteen years presented with the initial damages to the oral tissues and the distinctive long term effects in conjunction with the congenital analgia syndrome. The severity of this syndrome justifies the consideration of a prophylactic extraction of the primary dentition. A controlled mastication will be more likely with increasing age and eruption of the permanent teeth. PMID- 9328677 TI - In vitro susceptibility of Staphylococcus aureus including MRSA to four disinfectants. AB - The spread of nosocomial infections caused by pathogenic organisms such as methicillin-resistant S. aureus (MRSA) has prompted the dental community to focus more attention on certain control strategies. In the present study, we tested the abilities of the four skin disinfectants (povidone iodine, benzalkonium chloride, chlorhexidine gluconate, and ethanol) to prevent horizontal transmission of MRSA in the dental office. The bactericidal activities of the disinfectants were evaluated by the decrement over time of viable cell numbers of four clinical isolated strains of S. aureus: two MRSA strains and two methicillin-sensitive S. aureus (MSSA) strains. The most effective disinfectant was 70 percent ethanol, which eradicated both MRSA and MSSA in less than three minutes. The 0.1 percent chlorhexidine gluconate proved the least effective of four disinfectants. More than 10(2) bacteria survived despite exposure to it for thirty minutes. PMID- 9328679 TI - Treatment trends during a thirteen-year period in a student pediatric dentistry clinic. AB - This manuscript reports the treatment trends in a pediatric dentistry clinic from 1980 to 1992 and discusses their implication in clinical teaching. Analysis of the records of the senior year pediatric dentistry students indicated: no significant change with time in the patients/student ratio, the number of preformed crowns, pulpotomies, and pulpectomies by student or by patient; a significant decrease in the number of one-surface and > or = 2-surface restorations by student and by patient; a significant increase in the number of pit-and-fissure sealants and preventive resin restorations by student and by patient. During the thirteen-year period, the students performed an average of 7.3 one-surface; 12.9 > or = 2-surface restorations; 5.5 preformed crowns; 6.4 pit-and-fissure sealants; 2.4 pulpotomies. There was a significant increase with time in the number of students who performed pit-and-fissure sealants. PMID- 9328680 TI - Replantation of avulsed primary anterior teeth: treatment and limitations. AB - In addition to the successful replantation of avulsed permanent teeth, the replantation of primary anterior teeth may also be indicated. The decision is based on age and stage of tooth development, development of dentition, storage of the avulsed tooth and the way it is transported to the treatment site, the appropriate in vitro treatment of the tooth before reinsertion, and the willingness of the child to cooperate. A method involving retrograde filling of the primary tooth root with calcium hydroxide after resecting the root apex has proved successful. Other commercially available root filling materials and pins are not indicated. Calcium hydroxide allows the tooth to heal in place without reaction and prevents the development of apical periodontitis. As regards any surgical intervention, the attending dentist in this case has to weigh the benefits against the risks. PMID- 9328681 TI - Evaluation of mandibular infiltration versus block anesthesia in pediatric dentistry. AB - The clinical effectiveness of mandibular block anesthesia was compared to that of buccal infiltration anesthesia. A total of eighty patients three to nine years old was selected with identical bilateral lesions. The anesthetic used was mepacaine HCL 2 percent. The treatments performed were restorations, pulpotomies, and extractions. Child behavior and pain reaction were recorded and rated at certain intervals of treatment, using the Frankl behavioral scale and the SEM scale. The Eland color scale was also used in another sample of twenty patients to determine which type of anesthesia was more acceptable to children. The paired t-test was used to compare results. Buccal infiltration anesthesia was found to be as effective as block anesthesia in all situations, except when pulpotomies were performed in the mandibular second primary molar, where it proved to be unreliable regardless of age. Block anesthesia was significantly more painful than buccal infiltration anesthesia, and behavior of children three through five years old sometimes turned negative following the block injection. PMID- 9328682 TI - Child poverty vs Medicare and Social Security. AB - The series of government safety net programs provides economic security primarily to older populations. In this period of competition for limited federal resources, the need to create a public awareness of the continuing and growing poverty of children is emphasized. PMID- 9328683 TI - Raising children is expensive in the 1990s and beyond. AB - Costs of raising children continue to rise. At the same time, the marked increase in the proportion of two-earner families and single-parent families places added burdens on families to develop and pay for child care arrangements. A review is provided of current and anticipated financial costs of raising children during the 1990s and planning for the finances of their future years. PMID- 9328684 TI - Eosinophilic granuloma: report of case. AB - This paper related a case of eosinophilic granuloma in an eleven-year-old male child, treated successfully with curettage. The lesion appeared as a single irregular nodule in the left mandibular alveolar crest. A radiolucency was observed in the radiograph apparently causing an expansion of the vestibular and lingual cortical plates and displacement of the germs of the canine and first premolar. The treatment was by curettage, retaining the germs of the teeth involved. Other parts of the body were examined by osteal scintillography. Clinical and radiographic observations were done in the first three months; one year; two years; and five years postoperative with excellent prognosis. PMID- 9328685 TI - Gingival overgrowth with valproic acid: a case report. AB - A case of a nine-year-old epileptic girl with severe gingival overgrowth who had been taking valproic acid since two months of age is presented. A review of the literature and possible mechanisms for drug-induced gingival overgrowth is outlined. PMID- 9328686 TI - Anomalies of tooth form and number in the permanent dentition: report of two cases. AB - Two cases of bilateral double teeth involving the permanent maxillary central incisors are described. The difficulties in establishing the precise diagnosis are considered. The treatment plans also are discussed. The etiology and nomenclature of these dental formations and number of anomalies are reviewed. PMID- 9328687 TI - Redundant publication. AB - The following is a consensus statement from the Heart Editors Action Round Table Group concerning its policy on redundant publication. This statement is being published in journals represented on the panel beginning in July, 1997. PMID- 9328688 TI - Comparison of angiography and intravascular ultrasound for the assessment of lumen size after coronary stent placement: impact of dilation pressures. AB - This study was designed to assess the extent of potential discrepancies between intravascular ultrasound (IVUS) and quantitative coronary angiography (QCA) measurement of intrastent minimal luminal diameter and to evaluate the impact of dilation pressures and the balloon:artery ratio on the assessment of the minimal lumen diameter (MLD) by these imaging modalities. IVUS is recommended as an adjunct to angiography to assess stent expansion; however, the extent of potential discrepancies between the two imaging modalities is not well defined. Included were 225 patients in whom coronary Palmaz-Schatz stents were successfully placed after PTCA. IVUS and QCA were performed at the end of the intervention. We compared the MLD assessed by QCA and IVUS in the instent and reference site. The MLD assessed by IVUS and QCA were 2.68 +/- 0.41 mm and 3.08 +/- 0.47 mm (P < 0.001), respectively, at the tightest intrastent site and 3.19 +/- 0.50 mm and 3.17 +/- 0.52 ns at the reference site. There was a correlation between the dilation pressure and the difference between QCA- and IVUS-based intrastent MLD measurement (y = -0.05x + 1.11; r = -0.53; P < 0.0001). At low dilation pressures, a significant difference between the image modalities was found, but after high dilation pressures no discrepancies were detected. No relation was found with the balloon:artery ratio. These data provide clear evidence that in the case of low-pressure dilation, the exclusive reliance on data obtained by QCA will not yield sufficiently accurate information on intrastent MLD, whereas after high dilation pressure, the differences between the imaging modalities are minimized. PMID- 9328689 TI - "QCA and the Emperor's new clothes". PMID- 9328690 TI - Radiation exposure to patients undergoing diagnostic and interventional cardiac catheterization procedures. AB - There has been a recent wave of concern over radiation exposure to patients during cardiac catheterization. Accordingly, we measured the area-exposure product (AEP) to patients undergoing diagnostic and interventional cardiac catheterization procedures. Fluoroscopic and cinefluorographic exposures were determined for 510 patients using an AEP meter. The total AEP was higher in interventional than diagnostic cases (16,289 R.cm2 vs. 10,873 R.cm2, P < 0.00001); multi-lesion than single lesion interventions (20,311 R.cm2 vs. 15,919 R.cm2 P < 0.0001); and in patients with previous coronary bypass surgery (20,403 R.cm2 vs. 14,298 R.cm2, P < 0.00001). The highest AEPs were observed in patients with a prior history of bypass surgery who underwent diagnostic catheterization and multilesion intervention during the same procedure. PMID- 9328691 TI - Radiation exposure: clueless in the cath lab, or sayonara ALARA. PMID- 9328692 TI - X-ray-ted. PMID- 9328693 TI - One balloon approach for optimized Palmaz-Schatz stent implantation: the MUSCAT trial. AB - BACKGROUND: After stent deployment, larger balloons are frequently needed to optimize stent expansion according to angiographic and intravascular ultrasound (IVUS) criteria. The objective of this trial was to assess the feasibility and safety of a single-balloon approach for predilation, stent implantation, and optimization with a differential-compliant balloon allowing for focal overexpansion. We also evaluated the achieved degree of stent expansion according to IVUS criteria. METHODS AND RESULTS: Forty-seven consecutive patients with 50 lesions received single or multiple Palmaz-Schatz coronary stents. The final angiographic diameter stenosis was -2.6 +/- 12.6% (reference diameter, 2.89 +/- 0.44 mm), and the residual lumen area stenosis (IVUS) was 13.0 +/- 12.3% (reference area 10.8 +/- 3.0 mm2). This result was achieved in two steps (first angiographic, then IVUS-guided stent optimization). The balloon inflation pressure increased from 13.1 +/- 3.0 bar at step 1 to 16.1 +/- 3.0 bar at step 2, which resulted in a balloon to artery ratio of 0.97 +/- 0.12 and 1.10 +/- 0.15, respectively, at the low-compliant peripheral balloon segments. The more compliant central balloon segments showed a balloon to artery ratio of 1.09 +/- 0.17 and 1.28 +/- 0.17, respectively. The primary success rate for stent deployment was 94%. Acute complications included two type A and one type B dissection without clinical sequelae. CONCLUSIONS: The single-balloon approach for stenting is feasible and safe. The acute result is comparable to that of other studies with IVUS-guided stent optimization, the primary success rate, however, is slightly lower with the presently available catheter. PMID- 9328694 TI - Focal stenting: does it make sense? PMID- 9328696 TI - How to avoid cardiac tamponade during percutaneous balloon mitral valvuloplasty. PMID- 9328695 TI - Mechanisms of cardiac perforation leading to tamponade in balloon mitral valvuloplasty. AB - Mechanisms of cardiac perforation in 10 cases of cardiac tamponade encountered in a single-center series of 903 balloon mitral valvuloplasty procedures were elucidated by precise localization of the site of perforation at subsequent surgery. These mechanisms were perforation of the aortic root and adjacent right atrium by sliding up of the transseptal set (2), apical tears by straight-tip balloon catheters driven distally during mitral valve dilatation (3), apical perforations by guidewires introduced through catheters wedged in the apex (2), tear of the posterior right atrial wall by dilatation of the track produced by very low septal punctures (2), and right ventricular perforation by a pacing catheter (1). Multivariate analysis showed cardiac perforation to be significantly related to the total experience at the center (inversely) and to patient age (directly). Left ventricular perforation occurred exclusively in patients > 40 yr of age. Understanding these mechanisms has enabled formulation of effective strategies to prevent cardiac perforation. PMID- 9328697 TI - Cardiac perforation and tamponade: being at the wrong place but at predictable times during balloon mitral commissurotomy. PMID- 9328699 TI - Infarct angioplasty: the hole story. PMID- 9328698 TI - Primary angioplasty reduces risk of myocardial rupture compared to thrombolysis for acute myocardial infarction. AB - Although the mechanical complications of acute ventricular septal defect and acute mitral regurgitation are uncommon after acute myocardial infarction, these complications are associated with an extremely high morbidity and mortality. We hypothesized that the administration of thrombolytic drugs may result in hemorrhagic infarction as well as the potential for incomplete revascularization and thus may lead to an increased incidence of mechanical complications compared to primary angioplasty. Accordingly, we reviewed the data of the most contemporary thrombolytic and primary angioplasty trials and compared the incidence of mechanical complications among 36,303 patients treated with thrombolytics reported in the GUSTO trial to the incidence of mechanical complications among 1,295 patients treated with primary angioplasty obtained from the PAMI-1 and PAMI-2 trials. We found that angioplasty resulted in an overall 86% relative risk reduction in mechanical complications (2.20% vs. 0.31%, P < 0.001). In comparison to thrombolytic therapy, angioplasty resulted in an 82% decrease in acute mitral regurgitation (1.73% vs. 0.31%, P < 0.001) and a 100% decrease in acute ventricular septal defect (0.47% vs. 0.00%, P < 0.03). In conclusion, in patients with acute myocardial infarction, reperfusion with primary angioplasty is associated with less myocardial rupture and mechanical complications than thrombolytics. This finding may, in part, explain the improved prognosis observed in myocardial infarction patients treated with primary angioplasty. PMID- 9328700 TI - Multiple stent implantation in single coronary arteries: acute results and six month angiographic follow-up. AB - A total of 147 stents were implanted (in overlapping manner in 76% of vessels) in a single coronary artery in 59 patients (60 vessels, 97 lesions, 2.45 stents/vessel) over a period of 18 mo using high pressure stent deployment without ultrasound guidance. The indications for stenting were suboptimal percutaneous transluminal coronary angioplasty (PTCA) result (45%), primary prevention of restenosis (44%), acute closure (10%), and restenosis after plain balloon angioplasty (1%). One patient required emergency coronary artery bypass grafting (CABG) (extensive dissection), and one required early intervention with plain balloon angioplasty and intracoronary urokinase for stent thrombosis. There were no deaths. Thirteen patients had recurrence of angina within 6 mo and angiograms were performed in all. These showed intrastent restenosis in nine (all had successful repeat plain balloon angioplasty), development of new disease in other vessels along with restenosis close to the stent in the target vessel in one (underwent elective CABG) and normal angiograms with widely patent stents in three. Forty-five patients (77%) remained free of recurrent angina and 25 of these had follow-up angiograms (56%) at a mean of 172 days, two showing restenosis. Thus, the restenosis rate per patient in the symptomatic group (angiographic follow-up in 100%) was 77% and in the asymptomatic group (angiographic follow-up in 56%) was 8%. The restenosis rate in the subgroup with bailout stenting (n = 6) was 20% (angiographic follow-up in 83%). The overall restenosis rate per patient was 32% (overall angiographic follow-up in 66%). During the 6-mo follow-up period, one patient underwent elective CABG (1.7%), one sustained a non-Q myocardial infarction (1.7%), nine had repeat PTCA to the target vessel (15.5%), and there were no deaths. The event-free survival rate was 77%. Multiple stent implantation aided by high pressure stent deployment without ultrasound guidance and with adjunctive optimal antiplatelet therapy without oral anticoagulation seems to be a useful and effective revascularisation strategy to deal with long lesions and acute dissections with a high procedural success rate. The restenosis rate is acceptable and is not appreciably high as reported in previous studies from the "warfarin era." PMID- 9328701 TI - Initial experience with the Cordis stent: analysis of serial angiographic follow up. AB - To evaluate the efficacy of the more flexible Cordis stent, a prospective angiographic follow-up study was performed. Implantation of the Cordis stent was attempted in 99 consecutive patients with 103 native coronary lesions from January 1994 to July 1995. Clinical success, defined as final diameter stenosis of < 50% without death, bypass surgery, or Q-wave myocardial infarction, was achieved in 88% of the patients. There were no in-hospital deaths. In-hospital subacute stent occlusion occurred in only one case. Follow-up angiograms were obtained in 86 (95%) eligible lesions. The minimal luminal diameter improved from 1.03 +/- 0.31 to 2.82 +/- 0.31 mm, but started to decrease at 1 mon (2.57 +/- 0.24 mm), and continued to decrease throughout the 6 mon (2.00 +/- 0.61 mm), the biggest reduction being between 1 and 3 mon (-0.57 +/- 0.50 mm). Angiographic restenosis (stenosis > or = 50%) occurred in 23% of the lesions; a revascularization procedure of the target lesion was required in 12% of the patients. Multivariate analysis identified age, diabetes mellitus, and preprocedural reference diameter to be predictors of angiographic restenosis. In conclusion, the Cordis stent can be implanted successfully with a low complication rate and a clinical outcome at least comparable to other stent studies. PMID- 9328702 TI - Coronary artery stenting for suboptimal PTCA results in acute myocardial infarction in patients treated with Abciximab: early and six-month outcome. AB - Emergent percutaneous transluminal coronary angioplasty (PTCA) is an effective treatment for acute myocardial infarction. However, occasionally results of angioplasty are suboptimal due to coronary dissection or elastic recoil, leading to a high chance of recurrent ischemia. Coronary stents are occasionally employed in such settings, but a high incidence of stent thrombosis was noted by early investigators when stents were placed into areas of active thrombus formation. Since coronary thrombosis and stent thrombosis are both initiated by platelets, the potent antiplatelet agent abciximab might be useful in preventing stent thrombosis. Little information is available concerning early outcome or 6-month clinical event rate when coronary artery stents are placed for suboptimal angioplasty results for acute myocardial infarction in patients given abciximab. We deployed 75 stents as part of angioplasty for acute myocardial infarction in 40 patients given abciximab. All patients had suboptimal angioplasty results leading to stent deployment. Each obtained normal flow angiographically and no stent thrombosis or acute closure was observed. Early mortality occurred in 1 patient. All patients were followed at least 6 months, and no patient died after hospital discharge. Three patients experienced recurrent ischemic events within the first 6 months. Two of these events were due to infarct vessel restenosis. We conclude the combined use of coronary artery stents and abciximab for suboptimal PTCA results during acute myocardial infarction is associated with a low incidence of culprit vessel recurrent ischemic events within 6 months of intervention. PMID- 9328703 TI - Angioscopic evaluation of site-specific administration of ReoPro. AB - Systemic administration of newer antiplatelet agents such as the GP IIb/IIIa agent ReoPro (Centocor BV Leiden, The Netherlands) has been shown to decrease the early incidence of recurrent ischemia and recurrent myocardial infarction. The currently approved protocol for administration of ReoPro involves an initial weight adjusted bolus followed by a systemic infusion over the next 12 hr. The systemic administration is proposed as the only route of administration because it is stated that the drug must be exposed to circulating platelets. An alternative approach would be to deliver ReoPro locally and allow the platelets to disaggregate only when in contact with the local arterial wall. The optimal method of monitoring the efficacy of such a strategy is to visually assess the presence of platelet rich or red blood cell rich thrombus using angioscopy. We report our initial experience in 12 patients who received local administration of ReoPro using currently approved catheters for local administration of agents into coronary arteries who were evaluated before and after intervention using intracoronary angioscopy. None of the patients received a subsequent 12-hr infusion. There was successful resolution of thrombus in 11 of 12 patients. Recurrent ischemia occurred in one patient without myocardial infarction. PMID- 9328704 TI - Seeing is believing. PMID- 9328705 TI - Rapid reversal of no-reflow using Abciximab after coronary device intervention. AB - The no-reflow phenomenon is a reduction in epicardial coronary artery blood flow without mechanical vessel obstruction. Early descriptions of this syndrome involved reperfusion after myocardial infarction. More recently, the no-reflow phenomenon has been recognized after brief ischemic times associated with coronary interventions. It is clearly a negative prognostic indicator. The proposed mechanism is multi-factorial and may involve small vessel vasospasm and potentially platelet-mediated loss of capillary autoregulation. Because of the potential role of platelets in the genesis of the no-reflow state, we administered Abciximab to two patients with no-reflow phenomenon following catheter interventions. In both of these settings, rapid distal runoff was reestablished within minutes after treatment with the platelet glycoprotein 2B/3A inhibitor. PMID- 9328707 TI - Stenting for postoperative congenital heart disease in infants. PMID- 9328706 TI - Emergent stent placement for acute Blalock-Taussig shunt obstruction after stage 1 Norwood surgery. AB - A neonate underwent a stage 1 Norwood surgery for hypoplastic left heart syndrome and subsequently developed profound cyanosis and hemodynamic instability. Catheterization revealed an occluded modified Blalock-Taussig shunt. Angioplasty and stent implantation resulted in immediate angiographic and clinical improvement, which has persisted at 5-month follow-up. This therapy may provide lifesaving treatment in selected patients. PMID- 9328708 TI - Combined brachial and femoral approach to balloon angioplasty in coarctation of aorta. AB - Transfemoral balloon angioplasty of native aortic coarctation was initially not feasible in two patients, because of inability to cannulate the femoral artery percutaneously in one, and to cross the coarctation in the other. The percutaneous brachial approach helped overcome both these problems, after which utilization of intravascular snares allowed successful transfemoral completion of angioplasty. PMID- 9328709 TI - Concentric placement of stents to relieve an obstructed anomalous pulmonary venous connection. AB - A 9-month-old male with asplenia and complex congenital heart disease experienced progressive stenosis of an anomalous pulmonary venous connection. He developed pulmonary edema and growth failure. Two stents were placed concentrically to relieve the stenosis, and the pulmonary edema and growth failure resolved. Definitive surgery was accomplished 8 months later. PMID- 9328710 TI - Utility of magnetic resonance imaging in a patient with anomalous origin of the right coronary artery, acute myocardial infarction, and near-sudden cardiac death. AB - A 46-year-old female presented with an acute myocardial infarction and cardiac arrest. Coronary angiography revealed an anomalous origin of the right coronary artery coursing between the aorta and pulmonary artery. Magnetic resonance imaging confirmed the life-threatening nature of this anomaly and led to referral for surgical revascularization. PMID- 9328711 TI - Cardiac MRI: another imaging modality for coronary heart disease? PMID- 9328712 TI - Acute myocardial infarction caused by a myocardial bridge treated with intracoronary stenting. AB - The clinical significance of myocardial bridges (MBs) is variable, and most patients are asymptomatic. However, angina, myocardial infarction, and sudden death have been reported in association with MBs. Here we describe the use of intracoronary stenting for the treatment of a patient with an anterior myocardial infarction due to an MB. PMID- 9328713 TI - Restoration of patency of left internal mammary artery graft with progression of the underlying left anterior descending coronary artery disease. AB - The use of the left internal mammary artery (LIMA) to graft a borderline lesion in the left anterior descending coronary artery (LAD) has been associated with distal narrowing and occlusion of the LIMA. We present a patient in whom the LIMA occluded 1 year after coronary artery bypass, but was found to be fully patent 4 years later, after progression of the native LAD disease. PMID- 9328714 TI - Recurrent syncope due to complete atrioventricular block, a rare presenting symptom of otherwise silent coronary artery disease: successful treatment by PTCA. AB - A patient presented with recurrent syncope and episodes of AV block preceded by asymptomatic ST segment elevation on ambulatory monitoring. Coronary angiography revealed a severe stenosis in the midsegment of the right coronary artery (RCA). Successful PTCA and stent insertion abolished further episodes of syncope. PMID- 9328715 TI - Transjugular approach to transseptal balloon mitral valvuloplasty. AB - The feasibility of a transjugular approach to septal puncture and Inoue-balloon mitral valvuloplasty (BMV) was studied in 20 patients with severe mitral stenosis and varying degrees of anatomic atrial distortion. Left atrial entry by transjugular septal puncture was achieved without difficulty and BMV completed in all patients. In all of 16 patients who had high septal punctures, crossing the mitral valve with the Inoue-balloon was consistently simple and quick. In one patient, septal dilation after very high septal puncture led to a tear extending to the atrial free wall, resulting in cardiac tamponade requiring surgery. Another patient developed severe mitral regurgitation after BMV and required mitral valve replacement. Excellent results were obtained in 16 patients. The transjugular approach simplifies BMV procedure significantly in patients with distorted atrial anatomy and allows rapid patient mobilization. Its safety and efficacy need to be established in larger studies. PMID- 9328716 TI - A new view of an old picture. PMID- 9328717 TI - Novel technique for coil embolization of intercostal branch of internal mammary artery graft. AB - Coronary artery steal resulting from a large unligated intercostal or pericardial side branch of the internal mammary artery graft causing postoperative angina has been previously described. Transcatheter coil occlusion of internal mammary artery side branch has successfully been performed to treat coronary steal syndrome. Unsuccessful deployment of the microcoils can be due to inadequate guiding support in the LIMA or prolapse of the delivery catheter in the side branch. We report a new approach for the precise deployment of coils in the side branch of a LIMA graft, when inadequate guiding support is present. PMID- 9328718 TI - Vessel reconstruction by stenting of a long subintimal pathway. PMID- 9328719 TI - Time for a prospective national or international registry. PMID- 9328720 TI - New balloon expandable stent for bifurcation lesions. PMID- 9328721 TI - Periodic intermittent electromechanical dissociation: hemodynamic correlate of a malfunctioning mechanical prosthesis. PMID- 9328722 TI - Leaking balloon during stent deployment masquerading as coronary artery rupture. PMID- 9328723 TI - Management of hepatic venous outflow obstruction (Budd Chiari syndrome): balloon angioplasty with or without the use of a stent. PMID- 9328724 TI - Inoue balloon valvuloplasty for the tricuspid valve during pregnancy. PMID- 9328725 TI - "Seinfeld syncope". PMID- 9328726 TI - The molecular mechanisms of term and preterm labor: recent progress and clinical implications. AB - Current tocolytic protocols rely largely on the use of beta-mimetics to induce myometrial quiescence and delay delivery. Unfortunately, the rapid transplacental passage and poor receptor specificity of the commonly used beta-mimetics results in widespread activation of intrauterine and extrauterine beta 1 and beta 2 receptors. The use of beta-mimetics is associated with a range of well-recognized and potentially dangerous side effects for mother and fetus. The value of continued use of beta-agonists after obtaining uterine quiescence also has been the subject of recent debate. In this article we have attempted to explore the biochemical and molecular rationale for the use of alternative therapeutic modalities in the treatment and prevention of PTL. In the light of the current view that the term "preterm labor" covers a considerable diversity of causes, we propose that a range of treatment regimes should be chosen on the basis of the diagnosis and classification of the patient according to the their particular condition. Although the measurement of several biochemical parameters have been suggested to be of use in predicting PTL, we believe that a panel of diagnostic indicators (e.g., free or total CRH, IL-6, extracellular matrix proteases, fetal fibronectin) is more likely to provide useful diagnostic information on which appropriate treatment modalities can be selected (Table 1). Because of the complex and interactive nature of the mechanisms operating within the intrauterine environment to regulate myometrial activation and uterotonin production, we speculate that a combination of tocolytics, anti-inflammatory agents, uterotonic antagonists, and receptor blockers is likely to be more effective than a monotherapeutic approach, which focuses on only one facet of the process of uterine activation for pharmacologic intervention. For example, the use of antibiotics, PGHS inhibitors, and/or beta-mimetics might be an appropriate first line of treatment for infection-associated PTL in extreme prematurity. If it is successful, this treatment might be followed by longer term use of a progestagen and/or NO donor to maintain myometrial quiescence until closer to term. Alternatively, use of progesterone or oxytocin antagonists may be effective in augmenting the actions of beta-mimetics while reducing their side effects, whereas other combinations may be useful as long-term prophylactics in women with a high risk of developing PTL. Improvements continue in our diagnostic ability to correctly identify the different causes of preterm labor. We anticipate that careful selection of differing combinations of therapeutic options will result in significant reductions in the morbidity, mortality, and healthcare costs associated with preterm birth. PMID- 9328727 TI - The genetics of labor. PMID- 9328728 TI - Induction of labor. PMID- 9328729 TI - Elective induction of labor. PMID- 9328730 TI - Active management of labor: the American experience. PMID- 9328731 TI - Oxytocin: use and abuse, science and art. PMID- 9328732 TI - Managing difficult labor: avoiding common pitfalls. AB - The suggestions offered in this article represent an effort to reduce the incidence of cesarean delivery for dystocia while maintaining a safe course to vaginal birth. Avoiding difficult labor induction in which a compelling indication is lacking, providing prompt and effective oxytocin therapy of arrested first stage labor, and liberalizing the use of oxytocin therapy in selected cases of second-stage arrest are emphasized. With the widening use of conduction analgesia, indicated operative vaginal delivery has an increasingly important role in tempering cesarean birth rates. Operative vaginal delivery can play an effective role only when strict conditions to insure its safety are met. PMID- 9328733 TI - Avoiding labor problems during vaginal birth after cesarean delivery. PMID- 9328734 TI - Using evidence-based medicine to optimize cesarean section outcomes. PMID- 9328735 TI - Labor and delivery: a patient's perspective. PMID- 9328736 TI - Office assessment of chronic pelvic pain. AB - Discussion after the completion of the history and pelvic examination should center on education of the patient and her family about the multifactorial nature of chronic pain, and hence the necessity to use multiple treatment methods. The previously described detailed nature of the examination will help the clinician in directing further diagnostic and treatment efforts, as well as in making referrals. PMID- 9328737 TI - Premenstrual syndromes. AB - Premenstrual syndrome research has made a great deal of progress since 1983 when the criteria for the diagnosis were clearly defined. Confirming the diagnosis prospectively and ruling out other disorders was a major methodologic advance. The DSM-IV criteria for PMDD now help us identify and classify women who have severe psychologic symptoms during the premenstruum. Although we do not have a definitive cause for PMDD, the consensus is that it is the end result of a complex series of events mediated partly by the serotonin system and triggered by ovulation. Women who meet criteria for PMS, do not meet the criteria for PMDD, and do not have a concurrent disorder should be treated conservatively. Women who meet criteria for PMDD can be treated successfully with low-dose clomipramine, SSRIs, or GnRH-as with "add back" estrogen and progestins. PMID- 9328738 TI - Depression in women. PMID- 9328739 TI - Medically unexplained physical symptoms. PMID- 9328741 TI - Evaluating sexual dysfunction in women. PMID- 9328740 TI - Eating disorders. PMID- 9328742 TI - Domestic violence: a public health crisis. PMID- 9328743 TI - Marital counseling. PMID- 9328744 TI - Sexual assault. AB - All patients should be screened for a history of sexual abuse and victimization. Survivors of acute sexual assault have physical, psychological, and legal needs. The goal is to minimize additional trauma while simultaneously ensuring quality care and maximizing efforts to collect evidence. A pertinent history, physical examination, and treatment plan should be completed. Chain of custody needs to be maintained. The patient should be discharged with family or friends with appropriate follow-up. PMID- 9328745 TI - Helping patients cope with infertility. PMID- 9328746 TI - Psychosocial aspects of induced abortion. PMID- 9328747 TI - Women in the workplace. PMID- 9328775 TI - The challenge of the "difficult patient" (deja vu all over again--only more so) PMID- 9328776 TI - Predictors of physician frustration in the care of patients with rheumatological complaints. AB - Recent studies of the doctor-patient relationship have shown that certain patients are perceived as frustrating or difficult by their doctors; however, little is known about the characteristics of these patients that elicit this dissatisfaction. As part of a larger study of rheumatology clinic patients with fibromyalgia or rheumatoid arthritis (N = 68) we used stepwise multiple regression to select the factors most associated with physician frustration while controlling for the effects of other variables. Variable domains included demographics, psychiatric diagnoses, personality factors, functional disability, disease state, and trauma history. These domains as well as individual variables within these domains were systematically evaluated for their unique contribution to the prediction of physician frustration as measured by the Difficult Doctor Patient Relationship Questionnaire (DDPRQ). Initial bivariate correlates of physician frustration included marital status, current dysthymia and agoraphobia, lifetime panic disorder and obsessive-compulsive disorder, adult rape and physical abuse, somatization disorder, physical and social disability, the presence of fibromyalgia, as well as neuroticism, illness impact, and perceived loss of control. The best multivariable model for estimating frustration magnitude included somatization disorder, perception of lack of control over illness, and a lifetime history of obsessive-compulsive disorder. These factors explained 48% of the variance in DDPRQ score. Physicians in this study were most frustrated with patients who had ongoing preoccupation with multiple medically unexplained physical symptoms as well as the perception of greater impact and lack of control over their illness. These findings suggest that treatment of somatization in patients with chronic symptoms may decrease physician frustration. PMID- 9328777 TI - Integrating mental health services within primary care. A Canadian program. AB - The increasingly prominent role of the family physician in delivering mental health care can be enhanced if productive and collaborative relationships can be established with local mental health services. This paper describes a Canadian program that has achieved this by bringing mental health counselors and psychiatrists into the offices of 87 family physicians in 35 practices in a community in Southern Ontario. The paper describes the program, the activities of counselors and psychiatrists within the practices, and the administrative structures set up to coordinate these activities. Data is presented from the evaluation of the first year of the program's operation (13 practices and 45 family physicians) during which time 3085 referrals were received. The program made mental health care more available and accessible, increased continuity of care, provided additional support for the family physician, offered new opportunities for continuing education, and led to a reduced and more efficient use of other mental health services. The components of the program can be adapted to most communities. PMID- 9328778 TI - Short-term outcomes of detected and undetected depressed primary care patients and depressed psychiatric patients. AB - The aims of this study were to determine whether detection of major depression in primary care was associated with improved outcome, and to compare the 4.5 month outcomes of detected and undetected depressed primary care patients and depressed psychiatric patients. Primary care patients with major depression were recruited from the practices of 50 family physicians in Southeastern Michigan using a two stage selection procedure employing the Center for Epidemiologic Studies Depression Scale (CES-D) and the Structured Clinical Interview for DSM-III-R (SCID); clinician detection of depression was ascertained by response to a direct query on a rating form. Depressed patients seeking treatment in an outpatient psychiatric setting also received the CES-D and the SCID. Data on patient demographics and clinical characteristics were obtained for both primary care and psychiatric patients. Initial and 4.5 month scores on the Hamilton Depression Rating Scale (HAM-D) were obtained for 34 undetected and 25 detected depressed primary care and 55 depressed psychiatric patients. Improvement in depression over time was assessed by the change in HAM-D scores over the 4.5 months. The three groups did not differ in initial severity. Both psychiatric and undetected primary care patients showed significant improvement at 4.5 months, whereas detected primary care patients did not improve. At 4.5 months there were no differences in mean HAM-D scores between undetected, depressed primary care patients and depressed psychiatric outpatients. This result did not change after controlling for age and severity of depression at initial presentation, nor did it change after exclusion of cases of mild depression to control for a possible "floor effect." However, differences among groups in the stage of depressive episodes may have affected this comparison. These findings suggest that an exclusive focus on increasing detection of depression in primary care patients is unlikely to improve outcomes, and that undetected depression among primary care patients does not necessarily represent poor quality of care. Although depressed psychiatric patients in this study had better outcomes than detected depressed primary care patients, the presence of unmeasured differences among groups in the stage of the depressive episode makes it impossible to determine whether treatment of depression by psychiatrists is superior to that provided by primary care physicians. These findings should stimulate efforts to examine a more comprehensive model for detection and treatment of depression in primary care. PMID- 9328779 TI - The diagnosis and treatment of subclinical hypothyroid states in depressed patients. AB - The diagnosis and treatment of subclinical hypothyroidism in mood-disordered patients is complex and somewhat controversial. The psychiatrist must recognize that such subclinical states may contribute to depression, mood cycling, or delayed response to treatment if undetected. Autoimmune thyroiditis, which may ultimately lead to various grades of hypothyroidism, may also be seen in depressed populations, particularly in postpartum females. The author discusses these issues by means of a clinical vignette, then proposes an algorithm for the diagnosis and treatment of hypothyroid states in mood disordered populations. PMID- 9328780 TI - Factors associated with unplanned discharge from psychiatric day treatment programs. A multicenter study. AB - This study describes and evaluates the adequacy of sociodemographic and clinical descriptors as potential predictors of unplanned discharge from and completion of psychiatric day treatment programs. A 2-year retrospective chart review was completed on all patients (N = 327) attending three university-affiliated day treatment programs. Statistical comparisons were made between those patients who completed and those who had unplanned discharge. Logistic regression was used to generate a predictive model of unplanned discharge using identified variables. The rate of unplanned discharge was 54%. Factors associated with program completion were diagnoses of major depression or posttraumatic stress disorder, a history of completing a prior day treatment program, and higher education levels. Active substance abuse and a history of three or more inpatient admissions were associated with unplanned discharge. The predictive model was able to correctly classify 71% of patients completing day treatment programs and 43% of patients with unplanned discharges. Traditional demographic and clinical variables contribute differentially to program completion/noncompletion. Given the relatively poor predictability of unplanned discharge in this study, patient selection practices based on these factors alone may be somewhat limiting. Interactive effects of patient and program characteristics need to be addressed to improve program outcome. PMID- 9328781 TI - Comparison of child psychiatric patients in hospital and community clinics in Hong Kong. AB - This is a prospective study comparing a consecutive sample of child psychiatric patients at a community child mental health clinic (N = 56) and hospital clinic (N = 42) in Hong Kong. The subjects and their parents were studied with standardized questionnaires and semistructured interviews at their first visits to the clinics. A review of the treatment received was conducted 15 months later. Across settings, subjects were similar in sociodemographic profile, degree of social adversity, number of preceding life events, duration of chief complaints, maternal psychopathology, parental explanatory models of the child's problems, and expectations in treatment. Community clinic attenders had more disruptive behavioral problems, received shorter treatment, and less inpatient care than hospital clinic attenders. Subjects in both settings had very similar previous help-seeking behaviors. The findings suggested that the community clinic attracted disturbed children of similar backgrounds. The community child mental health clinic appeared to be a viable alternative in providing psychiatric care to children in Hong Kong. PMID- 9328783 TI - Cultural factors in the treatment of a catatonic Chinese patient. PMID- 9328782 TI - Hypokalemia leading to torsades de pointes. Munchausen's disorder or bulimia nervosa? AB - A 45-year-old woman traveled over 1000 miles from a major metropolitan area to obtain another opinion for medically refractory diarrhea. She had an extremely complicated medical history with no outside records or family members accompanying her to give collateral history. She had multiple previous diagnostic evaluations including 13 surgical procedures and many therapeutic trials of various medications. She acknowledged a preoccupation with weight and appearance, described previous attempts to diet, and repetitively denied purging, including laxative abuse. During her hospitalization she had two episodes of torsades de pointes requiring cardiac defibrillation. Laboratory testing revealed hypokalemia at the time of these events, and a toxicology screen was positive for bisacodyl, confirming laxative abuse. When confronted by a combined team of cardiology, gastroenterology, and psychiatry specialist, she admitted her laxative abuse and surrendered her supply of Dulcolax tablets. The discussion addresses the procedures employed to detect her surreptitious medication use, the near lethal cardiac complications, and the appropriate psychiatric diagnosis. PMID- 9328784 TI - The appearance of the CD4+CD8+ phenotype on activated T cells: possible role of antigen transfer. AB - Stimulation of peripheral blood lymphocytes (PBL) with phytohemagglutinin (PHA) strikingly increased the proportion of CD4+CD8+ cells. Highly purified CD4+ and CD8+ lymphocyte populations cultured in the presence of PHA consistently failed to coexpress the CD8 and CD4 markers. Similarly, exposure of highly purified CD4+ cells to PHA and recombinant interleukin-2 resulted in augmented expression of CD25 but failed to induce the expression of CD8. When purified preparations of either CD4+ or CD8+ cells were activated separately for 3 days and incubated together for an additional 5 h, a considerable proportion of CD4+CD8+ cells was found in the mixture. Cycloheximide treatment did not prevent the appearance of the CD8 marker on CD4 cells. CD4+CD8+ cells isolated from PBL exposed for 3 days to PHA lost their CD8 antigenicity within 24-48 h in the absence of PHA. Increased levels of soluble CD4 and CD8 antigens were found in supernatant fluids of PHA-stimulated cells. T cells failed, however, to bind soluble markers even after prolonged incubation in the presence of supernatant fluids. Our studies show that activation of CD4+ cells per se does not elicit the CD4+CD8+ phenotype and that soluble T cell markers do not bind to T cells. Rather, it seems that direct cell-cell contact is required for the transfer of CD8 molecules from CD8+ cells to the membrane of CD4+ cells. PMID- 9328785 TI - TCRB clonotypes are present in CD4+ T cell populations prepared directly from rheumatoid synovium. AB - The identification of clonal T cells at sites of inflammation is hampered by the large number of polyclonal T cells that nonspecifically accumulate. In this report, we combine the use of T cell sorting with spectratyping of the third complementarity determining region (CDR3) and direct sequence analysis to rapidly screen for and identify clonal expansions of T cells from synovial tissue specimens from patients with rheumatoid arthritis (RA). Initially, we used a polymerase chain reaction specific for the variable region gene of the T cell receptor beta chain (TCRBV) to compare the TCRBV repertoire expressed by CD4+ T cells from the peripheral blood and synovium of five patients with long-standing RA. Each patient had several TCRBV genes that were amplified to a greater degree from synovium. Extensive sequence analysis (n > 170) showed that each patient contained junctional sequences that occurred more than once, implying the presence of T cell clones within the starting CD4+ T cell population. To assess a more straightforward approach to identifying clones, six additional patients were recruited and CD4+, TCRBV2+ synovial T cells were positively selected and analyzed by CDR3 spectratyping. Bands deviating from a normal distribution were excised from the gel and sequenced directly. Clones were detected in half of the patients. These data are consistent with the possibility of an antigen-driven T cell response in RA that remains present in the setting of advanced disease. PMID- 9328786 TI - Identical T-cell receptor beta chain rearrangements are present in T cells infiltrating the jejunal mucosa of untreated celiac patients. AB - The intestinal mucosal lesion in celiac disease is characterized by a predominant T-cell infiltration of both epithelium and lamina propria. However, a restricted use of T-cell receptors (TCR) in T lymphocytes infiltrating the jejunal mucosa of celiac patients has not been reported. Based on an immunohistochemical survey of jejunal biopsies from a cohort of untreated celiac patients, we demonstrated a small but significant increase of V beta 8.1/2+ T cells in the lamina propria, but not in the epithelium nor in the peripheral blood. Sequence analysis indicated the existence of a variable degree of clonality of V beta 8+ T cells in the celiac mucosa. More importantly, the recurrence of identical CDR3 regions in some patients was also observed. The altered distribution of V beta 8+ T cells and the presence of identical CDR3 regions in celiac patients, but not in controls was independently confirmed by CDR3 size analysis in a further cohort of patients. These findings suggest that disease-specific variations of the TCRBV8 repertoire are present in the small intestinal mucosa of untreated celiac patients. PMID- 9328787 TI - Peptide recognition by T-cell clones of an HLA-DRB1*1501/*0901 heterozygous donor is promiscuous only between parental alleles. AB - The HLA class II isotype and allelic restrictions of peptide recognition were analyzed with T cells from a DRB1*1501/DRB1*0901 heterozygous donor. Nineteen T cell clones, all directed against the single mycobacterial epitope p21-40 were tested with HLA homozygous lymphoblastoid cell lines as antigen-presenting cells. The most striking finding has been, that several DR isotype restricted clones recognized the peptide in the context of both parental, but not of unrelated alleles. In contrast, DQ and DP restricted clones responded in the context of one parental allele only. Most DR promiscuous clones produced interferon-gamma but not IL-4, whereas most DQ and DP clones produced IL-4. We postulate that the confinement of DR promiscuity only to the parental alleles was established possibly during thymic maturation of T cells and that the proportions between monogamous and promiscuous T cells may play a role in the MHC mediated influences on host resistance to infections and other immune responses. PMID- 9328788 TI - Defective expression of CD95 (FAS/APO-1) molecule suggests apoptosis impairment of T and B cells in HLA-B8, DR3-positive individuals. AB - Activation-induced apoptosis is one of the primary control mechanisms for the negative selection of an immune response, leading to maintenance of immune homeostasis and selective T cell deletion. The interaction between the surface molecule Fas and its ligand (FasL) has been proposed as a primary mechanism initiating T cell apoptosis. The T cell receptor modulates the expression and function of these molecules. Defects in the Fas/FasL apoptosis pathway have been shown to result in autoimmune disease in humans and in murine models. Because subjects carrying the HLA-B8, DR3 haplotype show a number of immune dysfunctions, including membrano-proliferative glomerulonephritis, systemic lupus erythematosus, Graves' disease, and others, we investigated Fas expression on T and B cells, and sensitivity to Fas-mediated apoptosis of activated T cells, to determine whether abnormalities of the Fas pathway might be associated with the development of autoimmune diseases in this group of individuals. Our findings show that B cells and resting T cells from HLA-B8+, DR3+ subjects express markedly reduced levels of Fas compared with those isolated from HLA-B8-, DR3+ individuals. Reduced levels of Fas were also evident on the surface of T cells from HLA-B8+, DR3+ subjects activated in vitro by stimulation with OKT3 and phytohemoagglutinin. Cycling T cells from these subjects, evaluated for apoptotic nuclei by flow cytometry after incubation with a cytolytic anti-Fas mAb, showed a significantly lower percentage of Fas-mediated apoptosis than did those from HLA B8-, DR3- individuals. Normal levels of apoptosis were restored after exposure to a synthetic ceramide analog (C2). Further elucidation of the interaction of these molecules in autoimmune diseases may lead to better understanding of the pathogenesis of these disorders. PMID- 9328789 TI - HLA-DQ alleles associate with cutaneous features of onchocerciasis. The Kaduna London-Manchester Collaboration for Research on Onchocerciasis. AB - Onchocerciasis is associated with a spectrum of cutaneous changes, ranging from clinically normal skin to acute and chronic pathology. An important aspect of disease expression may be the level of immune response to parasite antigens, which is likely to be regulated by MHC-encoded molecules. We therefore investigated HLA class I and class II phenotypes in Nigerian residents of an area endemic for onchocerciasis. All study subjects were carefully characterized for parasite load and skin pathology. Individuals with depigmentation had increased frequencies of DQA1*0501 and DQB1*0301 compared with persons with normal skin and high microfilarial load (NSHMF) (Odds Ratios 3.6 (95% CI 1.0 to 13.2) and 3.8 (1.0 to 15.2), respectively). Conversely, individuals with depigmentation had a decreased frequency of DQA1*0101 and Cw6 compared with NSHMF (Odds Ratios 0.2 (0.1 to 0.9) and 0.1 (0.02 to 0.8), respectively). When NSHMF subjects were examined by age, a further decrease in DQA1*0501 frequency and increase in DQA1*0101 frequency were observed in older NSHMF individuals. These results strongly suggest that there is an immunogenetic basis for the spectrum of cutaneous presentations in onchocerciasis and that HLA-DQ molecules are associated with the level of immune response to parasite antigens. PMID- 9328790 TI - HLA class I and class II allele and haplotype distribution in the Venezuelan population. AB - Population studies represent an integral part, and a necessary link in a complex chain of host-pathogen interactions, disease pathogenesis, and major histocompatibility complex polymorphism. HLA class I and class II allele and haplotype distributions among Venezuelan mestizos were determined. Genes of Mongoloid, Negroid, and Caucasoid origin have created a distinctive human leukocyte antigen (HLA) genetic profile in this hybrid mestizo population that will influence HLA and disease association studies. The predominant HLA-B DQA1 DQB1 DRB1 haplotype is HLA-B44 DQA1*0201 DQB1*0201 DRB1*0701 (5.3%). It is noteworthy that the HLA-A3 B7 DR2 and the HLA-A1 B8 DR3 linkage groups, which are part of conserved or ancestral haplotypes, the last one associated with a wide range of autoimmune diseases and immune abnormalities in apparently healthy subjects, show low incidence among Venezuelan mestizos. This fact may be useful for future HLA and disease association studies and for localization of genes involved in immune regulation associated with these haplotypes. PMID- 9328791 TI - HLA-DR and -DQ associations with multiple sclerosis in Turkey. AB - The DRB, DQA, and DQB subregions of the major histocompatibility complex (MHC) were investigated by polymerase chain reaction and sequence-specific oligonucleotide probe hybridization (PCR/SSO) in 103 multiple sclerosis (MS) patients and 101 healthy controls from Turkey. Significant differences were detected between MS and control populations in the frequencies of DRB1*1501 [29 vs. 14, p = 0.02, odds ratio (OR) = 2.4], DRB1*04 (35 vs. 18, p = 0.01, OR = 2.3), DQB1*0302 (30 vs. 15, p = 0.02, OR = 2.3), DQB1*0602 (27 vs. 10, p = 0.005, OR = 3.2), DQB1*0501 (10 vs. 24, p = 0.01, OR = 0.3), DQA1*0101 (16 vs. 31, p = 0.02, OR = 0.4), and DQA1*0103 (7 vs. 19, p = 0.02, OR = 0.3). These results confirm the proposed positive association of the Dw2 (DRB1*1501 DQA1*0102 DQB1*0602) haplotype with MS in Caucasians in our Turkish population (25 vs. 8, p = 0.003, OR = 3.7). Furthermore, the "putative" haplotype supposed to be more frequent in the MS population of Mediterranean countries, namely DRB1*04 DQA1*03 DQB1*0302, is also associated with MS in Turkey (29 vs. 12, p = 0.006, OR = 2.9). The presence of two different haplotypic associations in MS emphasizes the complexity of the genetic susceptibility to MS in different populations. PMID- 9328792 TI - Polymorphism at codons 114, 116, 145, and 163 muddle the typing of HLA-B*1304. AB - Genetic exchanges often muddle the typing of HLA class I molecules, this is also the case for HLA-B*1304. Serologic and molecular DNA class I typing report a B15/B55 type for cell 847, whereas DNA sequencing finds B*5501/B*1304. HLA-B*1304 differs by no more than four amino acids from other HLA-B13 molecules, a comparative analysis of the B13 and B15 families was therefore performed to determine why serologic and molecular DNA approaches report a B15 type for B*1304. Comparisons demonstrate that limited differences individuate the B15 and B13 molecules such that the genetic recombination of codons 145 and 163 in the class I heavy chain's alpha 2 alpha helix prompt B*1304 to exhibit a B15X21 pattern of serologic cross-reactivity. Molecular DNA class I typing approaches are also swayed by genetic recombinations to type B*1304 as a B15 molecule: B15 like nucleotide sequences encoding residues 114, 116, and 145, lead B*1304 to exhibit a B15 PCR amplification pattern. Thus, genetic exchanges encoding key amino acids in the class I heavy chain lead molecular and serologic typing approaches to categorize HLA-B*1304 as a member of the B15 family. PMID- 9328794 TI - Nomenclature for factors of the HLA system, update May/June 1997. The WHO Nomenclature Committee for Factors of the HLA system. PMID- 9328793 TI - A comparative study of HLA-DRB typing by transcription-mediated amplification with the hybridization protection assay (TMA/HPA) versus PCR/SSOP. AB - To evaluate alternative human leukocyte antigen (HLA)-DNA typing methods, we used a system of transcription-mediated amplification (TMA) with a probe hybridization protection assay (HPA) in a microtiter plate format developed by Chugai Pharmaceutics Ltd. (Tokyo, Japan) to perform intermediate-level DRB typing for 502 individual samples. Two hundred fifty-two samples submitted to our Clinical Immunogenetics Laboratory were prospectively tested concurrently with a locally developed intermediate-level DRB polymerase chain reaction/sequence-specific oligonucleotide probe (PCR/SSOP) assay in a double-blind fashion. In addition, 250 retrospective samples of archived frozen cells or DNA from clinical and research panels, previously typed by allele-level DRB1 PCR/SSOP, were chosen to include 66 distinct DRB1 alleles representing Caucasian, American Black, Asian, and Native American ethnic groups. Among the prospectively typed samples, except for four samples with a TMA/HPA microplate handling problem, a single TMA/HPA allele assignment (1/462 alleles = 0.2%) was discordant with PCR/SSOP. Among the 250 retrospective samples, a single HPA probe for codon 57 aspartic acid consistently cross-reacted with the codon 57 valine sequence of DRB1*0807. However, TMA/HPA identified six samples with previous PCR/SSOP typing errors, all of which involved identification of sequences at codons 67-71 in samples heterozygous for two DR52-associated DRB1 alleles. Assay turnaround time from sample preparation to results was 11 h for 24 samples or 6-7 h for 1-4 samples. In summary, we found the TMA/HPA DRB typing system to provide rapid, reliable, and accurate HLA-DRB typing results. The current TMA/HPA methodology could be improved by use of a molded plastic cold block to provide more consistent and secure microtiter plate cooling than the current water/ice slurry. Nevertheless, this methodology, based on a microtiter plate format but without the usual plate washing steps of the traditional ELISA, has superior potential for microplate handling and reagent distribution with a robotics system and a work surface incorporating microplate heating and cooling units. PMID- 9328795 TI - Doxazosin suppresses the morning increase in blood pressure and sympathetic nervous activity in patients with essential hypertension. AB - To investigate the effects of doxazosin on blood pressure and sympathetic nervous activity, we analyzed the circadian variation of blood pressure and the power spectrum of R-R intervals using an ambulatory multibiomedical monitoring system (TM2425) in 10 untreated outpatients with essential hypertension. After a 2-wk placebo period (P-period), we administered 1 to 4 mg of doxazosin mesilate to the patients for 2 to 6 wk (T-period). We measured systolic and diastolic blood pressure (SBP, DBP), heart rate, R-R intervals, posture, and activity with the use of TM2425. Power spectral analysis of R-R intervals was used to calculate the ratio of low to high frequency components (LF/HF). The values were compared between the P-period and T-period. Although daytime blood pressure significantly decreased during the T-period (SBP, 148.1 +/- 5.9 vs. 130.3 +/- 4.4 mmHg; DBP, 92.3 +/- 3.2 vs. 83.6 +/- 2.6 mmHg, p < 0.01), nighttime DBP did not. The LF/HF of R-R intervals in the daytime (5.8 +/- 2.0 vs. 4.9 +/- 1.2, p < 0.01) and the morning rise in blood pressure also decreased significantly (SBP, 17.5 +/- 9.4 vs. 12.1 +/- 6.5 mmHg; DBP, 12.5 +/- 6.5 vs. 8.3 +/- 5.3 mmHg, p < 0.05). We conclude that doxazosin may suppress the morning rise in blood pressure in association with a decrease in sympathetic nervous activity. PMID- 9328796 TI - Circadian variation of hemodynamics and baroreflex functions in patients with essential hypertension. AB - It is well known that cardiovascular accidents such as myocardial infarction frequently occur in the morning, but their triggering mechanisms are not clear. The present study investigated circadian variations of hemodynamics and baroreflex functions. Twenty-three patients with essential hypertension were studied. Direct blood pressure (BP) and ECG were recorded by telemeter over 24 h, and then computer-analyzed. The pulse-contour method was used to measure cardiac output (CO) and total peripheral vascular resistance (TPR). The ratio of low to high frequency components (LF/HF) of the RR-interval on ECG was calculated by power spectral analysis. The baroreflex sensitivity index (BRI) was measured on the basis of the ratio delta RR/delta Ps (delta Ps = spontaneous decrease in systolic BP, delta RR = change in RR). Furthermore, 24-h BP changes were transformed algebraically into positive load component (PC) and negative load component (NC) by using a Windkessel model. The circadian variation of hematocrit (Ht) was also measured. The least squares method was used to determine the time at which the maximum and minimum value of each measurement occurred. Whereas the maximum values for BP and CO occurred in the evening (18:30, 17:00), the maximum values for TPR and LF/HF occurred between 06:30 and 08:00, and the minimum value for BRI occurred at 08:00. PC significantly correlated with Ps, heart rate, and CO (r = 0.81, 0.92, 0.67), and NC significantly correlated with BRI and LF/HF (r = 0.71, 0.64). PC (related to cardiovascular function) reached a maximum and NC (related to baroreflex function) reached a minimum in the late morning (11:00). Ht was highest immediately after the subjects got out of bed. These hemodynamic imbalances may negatively influence coronary blood flow in the morning. PMID- 9328797 TI - Predictive value of home blood pressure measurement in relation to stroke morbidity: a population-based pilot study in Ohasama, Japan. AB - We investigated the utility of home blood pressure measurements for determining the risk of stroke. We also analyzed the relationship between home blood pressure and the incidence of stroke. Home blood pressure and screening blood pressure measurements were obtained from 1,789 residents (aged 40 yr or older) of a rural Japanese community. Blood pressure was measured at home with a semiautomatic device. A mean (+/-SD) of 23.0 +/- 7.5 measurements were made for each subject. Subjects without a history of stroke and who were not receiving medication for hypertension (n = 1,256) were prospectively followed up for 4.4 +/- 2.1 yr. Subjects were subdivided into quintiles according to their baseline blood pressure. The association between the baseline blood pressure and the incidence of the first-ever stroke was examined with the Cox proportional hazards regression model, adjusted for age and sex. The lowest risk of stroke morbidity occurred in the subjects in the third quintile for home systolic blood pressure (117-123 mmHg) and in those in the second quintile for home diastolic blood pressure (66-70 mmHg). The subjects in the fifth quintiles for home systolic (> or = 133 mmHg) and diastolic blood pressure (> or = 81 mmHg) had a significantly increased risk of stroke morbidity. The subjects in the first and the second quintiles for home systolic blood pressure and those in the first quintile for home diastolic blood pressure tended to have an increased risk as compared with subjects in the lowest risk groups, although this increase was not statistically significant, indicating two possibilities: a trend toward a J-shaped relationship or no-decrease in risk of the first-ever stroke in subjects with home blood pressure level less than 123/70 mmHg. This relationship was not observed for screening blood pressure. We conclude that home blood pressure measurements can provide additional prognostic information to that obtained from blood pressure measurement in a medical environment. PMID- 9328798 TI - Ultrasonographic assessment of regional differences in atherosclerotic lesions in patients with hypertension, diabetes mellitus, or both. AB - We evaluated risk factors involved in regional differences in atherosclerotic lesions in patients with hypertension, diabetes mellitus, or both. Using ultrasonography, we examined the brachial, common carotid, and common femoral arteries in 65 hospitalized Japanese patients (15 controls, 18 patients with hypertension, 16 with diabetes mellitus, and 16 with both hypertension and diabetes mellitus). They ranged in age from 39 to 81 yr, mean 60.3 yr. The thickness of the intima-media complex of the far wall was measured, and the severity of atherosclerotic plaques was graded according to maximal lumen stenosis. The intima-media thickness in the carotid and femoral arteries was significantly greater in the hypertensive patients and the hypertensive patients with diabetes than in the controls. Severity of plaque was greater in the hypertensive patients with diabetes than in the controls. Plaque grades were higher in the carotid and femoral arteries than in the brachial artery. Multiple regression analysis revealed that age and mean blood pressure were strongly associated with the intima-media thickness in all three arteries. In the femoral artery, cigarette smoking and hyperglycemia also significantly correlated with the intima-media thickness. Plaque grades increased with age in the carotid and brachial arteries, while in the femoral artery the grade increased with cigarette smoking and serum cholesterol concentration. These findings suggest that the extent of atherosclerosis and its underlying risk factors differ among arterial sites. In addition, risk factors may partly differ according to the stage of atherosclerosis. To prevent or reverse atherosclerosis, the above differences should be taken into account. PMID- 9328799 TI - Cardiovascular effects of chronic inhibition of nitric oxide synthesis and dietary salt in spontaneously hypertensive rats. AB - The cardiovascular effects of chronic inhibition of nitric oxide synthesis and dietary salt were studied in 9-wk-old spontaneously hypertensive rats (SHR). N omega-nitro-L-arginine methyl ester (L-NAME, 0.025% in food, about 20 mg/kg/d) was given to rats receiving diets containing low, moderate, and high salt levels (NaCl 0.2%, 1.1%, and 6.0% of the dry weight of the chow) for 3 wk, L-NAME increased systolic blood pressure by 50 to 60 mmHg in all treated groups, as compared with an average rise of 10 to 20 mmHg in the control SHR. The high-salt diet did not further increase blood pressure. L-NAME also induced cardiac and renal hypertrophy, and these changes were aggravated by the high-salt diet. In addition, 19 of the 30 rats treated with L-NAME suffered strokes and all of them had several myocardial infarctions and renal damage, while the rats not treated with L-NAME had no evidence of stroke or myocardial or renal injury. Responses of mesenteric arterial rings in vitro were studied at the end of the experiment. The vascular contractile responses to noradrenaline were increased, and the relaxation responses to acetylcholine were inhibited in the L-NAME treated groups. In addition, the high-salt diet alone tended to inhibit the response to acetylcholine. Plasma renin activity was markedly increased by L-NAME treatment and decreased by the high-salt diet. The 24-h urine protein excretion was increased both by the L-NAME treatment and by the high-salt diet. The combination of L-NAME and the high-salt diet markedly raised the serum creatinine concentration. Our findings show that the coronary and renal functions are particularly vulnerable in SHR during impaired nitric oxide synthesis, and that the end-organ damage is worsened by an increased intake of dietary salt. We suggest that dysfunction of the endothelium is the primary cause of the effects observed in this study. PMID- 9328800 TI - Serum N-acetyl-beta-D-glucosaminidase activity in a genetic rat model of non insulin-dependent diabetes mellitus. AB - Serum N-acetyl-beta-D-glucosaminidase activity (NAG) is a possible predictor of vascular injury in hypertension. We assessed whether the activity of this enzyme reflects vascular damage in a genetic rat model of non-insulin-dependent diabetes mellitus (NIDDM) in humans. Otsuka Long-Evans Tokushima Fatty (OLETF) rats fed a regular chow were treated with the angiotensin converting enzyme (ACE) inhibitor imidapril for 16 wk. Systolic blood pressure increased in a time-dependent manner in the untreated OLETF rats as compared with that in the control Long-Evans Tokushima (LET) rats. The blood pressure elevation was associated with increases in cardiac and aortic weight. Imidapril treatment significantly attenuated the blood pressure elevation and reduced the increases in cardiac and aortic weight. The untreated OLETF rats had higher plasma glucose and insulin concentrations than did the LET rats and presented with glucosuria at the age of 22 wk. Imidapril treatment strikingly decreased plasma glucose levels and the glucosuria. Plasma insulin concentrations decreased, approaching those of the non diabetic control LET rats. ACE inhibitor treatment attenuated the nodular lesions in the glomeruli of OLETF rats and improved the kidney function. Serum NAG activity increased significantly by 35% in the untreated rats; this increase was attenuated significantly by imidapril treatment. The reduction in serum NAG activity correlated with improvement in cardiovascular injury. In contrast, there were no changes in urinary NAG excretion in the three OLETF rat groups. In addition, NAG excretion did not correlate with indices of cardiovascular injury. These data suggest that serum NAG activity is useful in predicting injury in the cardiovascular system in rats with diabetes mellitus. PMID- 9328801 TI - Preferential changes in hepatosplanchnic hemodynamics in patients with borderline hypertension. AB - To investigate changes in systemic and regional hemodynamics during the development of human hypertension, we simultaneously measured cardiac index (CI) by the indocyanine green (ICG) dye dilution method, hepatosplanchnic blood flow (HBF) by the ICG clearance method using a two-compartment model, and renal blood flow (RBF) by the p-aminohippurate clearance method in patients with borderline and essential hypertension. In patients with borderline hypertension (BH, n = 27), HBF (435 +/- 15 ml/min/m2) and HBF/CI (16 +/- 1%) were significantly (p < 0.05) lower than in age-matched normotensive controls (528 +/- 21 and 19 +/- 1, respectively, n = 21), while CI, RBF and RBF/CI were similar. In patients with essential hypertension (EH, n = 32), HBF, RBF, and RBF/CI were all significantly (p < 0.01) lower than in the control subjects. Hepatosplanchnic vascular resistance (HVR) in patients with BH was preferentially increased, while total peripheral resistance (TPR) and renal vascular resistance (RVR) remained in the normal range. In patients with EH, TPR, HVR, and RVR were all increased. These results indicate that hemodynamic changes in patients with BH do not occur uniformly among the various regional circulations and suggest that hemodynamic changes in the hepatosplanchnic region precede those in other organ circulations during the development of human hypertension. PMID- 9328802 TI - Arginine vasopressin inhibits nitric oxide synthesis in cytokine-stimulated vascular smooth muscle cells. AB - The purpose of this study was to investigate the effects of arginine vasopressin (AVP) on nitric oxide (NO) synthesis in vascular smooth muscle cells (VSMCs). We measured the production of nitrite, a stable metabolite of NO, and the expression of inducible NO synthase (iNOS) mRNA in cultured rat VSMCs. Incubation of VSMCs for 24 h with interleukin-1 beta (IL-1 beta) caused a significant increase in NO production. Both AVP and the V1a receptor agonist [Phe2, Ile3, Orn8]vasopressin inhibited NO synthesis in IL-1 beta-stimulated cells, but not in unstimulated cells, in a dose-dependent manner. The V1a receptor antagonist [d(CH2)5(1), O-Me Tyr2, Arg8]vasopressin completely inhibited the effect of AVP. Incubation with IL 1 beta for 24 h induced the expression of iNOS mRNA in VSMCs, while AVP suppressed its expression. After functional depletion of protein kinase C activity by treating cells with phorbol 12-myristate 13-acetate for 24 h, AVP did not inhibit IL-1 beta-induced NO production. The effect of AVP was also inhibited in the presence of the protein kinase C inhibitor calphostin C in a dose dependent manner. These results indicate that AVP inhibits IL-1 beta-induced iNOS expression in VSMCs through the V1a receptor, which is mediated at least partially via activation of protein kinase C. PMID- 9328803 TI - Stretch-induced proliferation of cultured vascular smooth muscle cells and a possible involvement of local renin-angiotensin system and platelet-derived growth factor (PDGF). AB - This experiment was designed to investigate the possible involvement of angiotensin II (Ang II) and platelet-derived growth factor (PDGF) in the mechanism underlying stretch-induced proliferation of vascular smooth muscle cells (SMCs). SMCs from the rabbit aortic media were grown on polystyrene rubber bottomed dishes coated with type I collagen. Cells were stretched cyclically by a vacuum-operated downward flexion of the culture dish bottom. A 1.4- to 1.6-fold increase in proliferation of SMCs was induced by cyclic stretching, as determined by [3H]-thymidine incorporation, in a stretch force-dependent manner in the range of 5% to 15% elongation, 30 cycles/min for 24 h. Expression of PDGF-B chain mRNA was up-regulated in a time-dependent manner in the range of 2 to 24 h, 10% elongation, and 30 cycles/min. Saralasin, a selective antagonist of Ang II, and captopril, an angiotensin I converting enzyme inhibitor, significantly suppressed the stretch-induced proliferation of SMCs. Blockade of angiotensinogen mRNA translation by antisense oligonucleotide inhibited proliferation under the mechanical strain. Stretch-induced proliferation was inhibited by 78% in the presence of anti-PDGF-AB neutralizing antibody. Increased expression of PDGF-B chain mRNA under the mechanical strain was inhibited by treatment with saralasin. Our results indicate that the stretch-induced proliferation of cultured SMCs is mediated at least in part via increased production of Ang II by the local renin angiotensin system and a subsequent up-regulation of PDGF-B chain mRNA in an autocrine-paracrine manner. PMID- 9328805 TI - The integration of pharmacological and nonpharmacological treatments in drug/alcohol addiction. PMID- 9328804 TI - Inhibitory effects of insulin on intracellular calcium and aggregatory response of platelets are impaired in hypertensive subjects with insulin resistance. AB - To determine the effects of insulin on intracellular calcium and platelet aggregatory responses in hypertensive subjects with insulin resistance, we measured insulin sensitivity in terms of glucose disposal rate (GDR) by the hyperinsulinemic euglycemic clamp technique (GC) in 14 non-diabetic untreated hypertensive subjects, and determined basal ([Ca2+]i) and thrombin-stimulated (T [Ca2+]i) platelet-free calcium concentrations and thrombin-stimulated platelet aggregatory response (AG) before (PRE[Ca2+]i, T-PRE[Ca2+]i, and PRE AG, respectively) and during (POST[Ca2+]i, T-POST[Ca2+]i, and POST AG, respectively) GC. As a control for GC, vehicle (normal saline) was infused on another day. No significant difference was observed between PRE[Ca2+]i and POST[Ca2+]i, T PRE[Ca2+]i and T-POST[Ca2+]i, or PRE AG and POST AG, GDR inversely correlated with delta[Ca2+]i (POST [Ca2+]i-PRE[Ca2+]i, r = -0.75, p < 0.02), delta T [Ca2+]i, (T-POST[Ca2+]i-T-PRE[Ca2+]i, r = -0.63, p < 0.02) and delta AG (POST AG PRE AG, r = -0.67, p < 0.01). No significant changes were observed in these variables during vehicle infusion. [Ca2+]i, T-[Ca2+]i, and AG decreased during GC as compared with baseline in hypertensive subjects with normal insulin sensitivity, but were unchanged in those with insulin resistance, suggesting that the vasodilatory and anti-platelet aggregatory effects of insulin are impaired in patients with insulin-resistant hypertension. PMID- 9328806 TI - Integration of generalized vulnerability to drug and alcohol addiction. AB - The vulnerability to develop addiction to alcohol has been well established in familial and genetic studies. Similar familial and genetic studies have supported a vulnerability to drug addiction. The co-occurrence of alcohol and drug addiction in the same individuals is highly prevalent in clinical populations. Common putative neurochemical mechanisms underlie addiction to both alcohol and drugs, namely, in the mesolimbic pathway and the locus ceruleus in the brain. Treatment strategies are directed at both alcohol and multiple drug addictions in patient populations. The formulation of a generalized vulnerability that extends to both alcohol and drug addiction is not only possible but necessary to explain the substantial numbers of individuals who develop both alcohol and drug addictions. Future research that is pertinent and relevant may depend on the understanding of a generalized vulnerability to develop alcohol and drug addiction and its application in diagnostic strategies and treatment models. PMID- 9328807 TI - Psychopharmacotherapy for addictive and comorbid disorders: current studies. AB - Proper diagnosis of comorbid disorders is crucial in treatment planning for the dually diagnosed. Since psychoactive substance use can obfuscate the diagnosis, special care must be taken to exclude organically based syndromes. Adequate periods of abstinence should first be achieved and subsequently the patient re examined for residual symptoms compatible with a nonaddictive, nonsubstance induced psychiatric disorder. The integration of concurrent treatment of both the mental and the addictive disorders appears to be the best approach for treatment of comorbid psychiatric and addictive disorders. An abstinence-based model that typically utilizes a 12-step group therapy is often employed for the addictive illnesses. Other forms of psychosocial therapies such as case managers are being used as well. Presently, physicians' prescribing practices for comorbid addicted patients are based on traditional approaches to use of medications in psychiatric patients, and their attitudes towards addictive disorders may play a significant role in determining the overall success of treatment. PMID- 9328808 TI - Integrating treatments for methamphetamine abuse: a psychosocial perspective. AB - The recent West Coast experience with increased methamphetamine use is showing signs of spreading to other parts of the US. The risk of corresponding medical and psychosocial problems has led to a call to action at the highest levels of government. The next few years will likely witness a substantial increase in treatment research on methamphetamine abuse/dependence, with particular emphasis on the development and application of novel pharmacotherapies. The evaluation of these agents presupposes that we understand the clinical syndrome resulting from chronic methamphetamine use. To establish a clear picture of the biological and psychological sequellae of methamphetamine use, we compare two cohorts (500 methamphetamine and 224 cocaine users) treated at the same outpatient clinic over the past nine years, using identical manualized treatments. The results suggest that while there are important differences in group characteristics and drug effects, the total response to treatment was quite comparable. PMID- 9328809 TI - The integration of pharmacotherapy and nonpharmacotherapy. PMID- 9328810 TI - Integration of pharmacotherapies in the existing programs for the treatment of alcoholics: an international perspective. AB - This review examines current therapies of alcoholism and addresses the difficulties encountered in evaluating the result of integrating pharmaco- and psychosocial treatment of alcoholics. A variety of treatment modalities and goals, and factors influencing admission of alcoholics, exist in different countries creating problems of comparability. Recognition of alcoholism as a separate entity from associated psychiatric co-morbidity coupled with the introduction of specific pharmacotherapies represent a major advance in the treatment of alcoholism. Combining psychosocial and pharmacological treatments appears to be the most effective approach in the treatment of alcoholics. In the future, rational therapy with appropriate choice of treatment strategies will decrease the necessity for hospital treatment of alcoholic patients. PMID- 9328811 TI - Integration of research in pharmacotherapy for addictive disease: where are we? Where are we going? AB - Systematic efforts by NIDA and NIAAA to develop new medications for drug and alcohol dependence have resulted in recent FDA approval of LAAM for opiate dependence, naltrexone for alcohol dependence and, more recently, a nasal spray for nicotine dependence. This article reviews the current strategies that guide these development efforts, including further examination of the interactions between potential pharmacotherapeutic agents and the drugs of abuse, the enhancement of pharmacotherapeutic efficacy with nonpharmacological interventions, and the development of more precise and meaningful measures of research outcome. PMID- 9328821 TI - New insights into the metastasis-associated 67 kD laminin receptor. AB - The interactions between tumor cells and laminin or other components of the extracellular matrix have been shown to play an important role in tumor invasion and metastasis. These interactions are mediated by different cell surface molecules, including the monomeric 67 kD laminin receptor. This molecule appears to be very peculiar since so, far only a full-length gene encoding a 37 kD precursor protein has been isolated and the mechanism by which the precursor reaches the mature form is not understood. Based on clinical data, which clearly demonstrate the importance of the receptor in tumor progression, studies were conducted to define the structure, expression, and function of this laminin receptor as a step toward developing therapeutic strategies that target this molecule. The data suggest that acylation of the precursor is the key mechanism in maturation of the 67 kD form. The function of the membrane receptor is to stabilize the binding of laminin to cell surface integrins, acting as an integrin accessory molecule, although homology of the gene encoding the receptor precursor with other genes suggests additional functions. Downregulation of the receptor expression on tumor cells might open new therapeutic approaches to decrease tumor aggressiveness. PMID- 9328822 TI - Prolidase activity in fibroblasts is regulated by interaction of extracellular matrix with cell surface integrin receptors. AB - Prolidase (EC 3.4.13.9) is a ubiquitously distributed imidodipeptidase that catalyzes the hydrolysis of C-terminal proline or hydroxyproline containing dipeptides. The enzyme plays an important role in the recycling of proline for collagen synthesis and cell growth. An increase in enzyme activity is correlated with increased rates of collagen turnover indicative of extracellular matrix (ECM) remodeling, but the mechanism linking prolidase activity and ECM is poorly understood. Thus, the effect of ECM-cell interaction on intracellular prolidase activity is of special interest. In cultured human skin fibroblasts, the interaction with ECM and, more specifically, type I collagen mediated by the beta 1 integrin receptor regulates cellular prolidase activity. Supporting evidence comes from the following observations: 1) in sparse cells with a low amount of ECM collagen or in confluent cells in which ECM collagen was removed by collagenase (but not by trypsin or elastase) treatment, prolidase activity was decreased; 2) this effect was reversed by the addition of type I collagen or beta 1 integrin antibody (agonist for beta 1 integrin receptor); 3) sparse cells (with typically low prolidase activity) showed increased prolidase activity when grown on plates coated with type I collagen or on type IV collagen and laminin, constituents of basement membrane; 4) the relative differences in prolidase activity due to collagenase treatment and subsequent recovery of the activity by beta 1 integrin antibody or type I collagen treatment were accompanied by parallel differences in the amount of the enzyme protein recovered from these cells, as shown by Western immunoblot analysis. Thus, we conclude that prolidase activity responded to ECM metabolism (tissue remodeling) through signals mediated by the integrin receptor. PMID- 9328823 TI - Insulin-like growth factor I inhibits the transcription of collagenase 3 in osteoblast cultures. AB - Insulin-like growth factor (IGF) I is an autocrine regulator of bone remodeling which inhibits bone collagen degradation and interstitial collagenase 3 mRNA levels. The mechanism of this inhibitory effect on collagenase 3 expression is not known. We tested the effects of IGF I on collagenase 3 gene expression in cultures of osteoblast-enriched cells from 22 day fetal rat calvariae (Ob cells) to determine whether transcriptional or posttranscriptional mechanisms were involved in the regulation of the collagenase 3 gene. IGF I at 10-100 nM caused a dose-dependent decrease in collagenase mRNA and protein levels. IGF I did not modify the half-life of collagenase 3 mRNA in transcriptionally arrested Ob cells, whereas it decreased the levels of interstitial collagenase 3 heterogeneous nuclear RNA. In addition, IGF I decreased the rates of transcription of the collagenase gene and the activity of a 2.1 kilobase collagenase 3 promoter construct transiently transfected into Ob cells. In conclusion, IGF I decreases the expression of collagenase 3 mRNA by transcriptional mechanisms. PMID- 9328824 TI - v-erbA oncogene initiates ultrastructural changes characteristic of early and intermediate events of meiotic maturation in Xenopus oocytes. AB - The growth-promoting properties of the retroviral v-erbA oncogene, a highly mutated version of the chicken thyroid hormone receptor (TR) alpha, have so far exclusively been linked to dominant repression of the antimitogenic roles of TR and retinoic acid receptors. Here we show that when expressed in Xenopus oocytes v-ErbA induced ultrastructural changes characteristic of early and intermediate events of meiotic maturation by activating gene transcription. v-ErbA-induced maturation events occurred without activation of the cAMP/maturation-promoting factor signal pathway and were arrested prior to meiotic spindle formation. The effects of v-ErbA were not mimicked by a dominant negative in vitro-generated mutant of human TR, suggesting that v-ErbA can contribute to cell cycle reentry by interference with regulatory pathways distinct from those involving TR. Interestingly, a portion of v-ErbA expressed in oocytes was present at the cytoplasmic fibrils of the nuclear pore complexes, suggesting that in addition to its intranuclear function v-ErbA may modulate nucleocytoplasmic transport. PMID- 9328825 TI - Reduced utilization of Man5GlcNAc2-P-P-lipid in a Lec9 mutant of Chinese hamster ovary cells: analysis of the steps in oligosaccharide-lipid assembly. AB - Recently we reported that CHB11-1-3, a Chinese hamster ovary cell mutant defective in glycosylation of asparagine-linked proteins, is defective in the synthesis of dolichol [Quellhorst et al., 343:19-26, 1997: Arch Biochem Biophys]. CHB11-1-3 was found to be in the Lec9 complementation group, which synthesizes polyprenol rather than dolichol. In this paper, levels of various polyprenyl derivatives in CHB11-1-3 are compared to levels of the corresponding dolichyl derivatives in parental cells. CHB11-1-3 was found to maintain near normal levels of Man5GlcNAc2-P-P-polyprenol and mannosylphosphorylpolyprenol, despite reduced rates of synthesis, by utilizing those intermediates at a reduced rate. The Man5GlcNAc2 oligosaccharide attached to prenol in CHB11-1-3 cells and to dolichol in parental cells is the same structure, as determined by acetolysis. Man5GlcNAc2 P-P-polyprenol and Man5GlcNAc5-P-P-dolichol both appeared to be translocated efficiently in an in vitro reaction. Glycosylation of G protein was compared in vesicular stomatitus virus (VSV)-infected parent and mutant; although a portion of G protein was compared in vesicular stomatitus virus (VSV)-infected parent and mutant; although a portion of G protein was normally glycosylated in CHB11-1-3 cells, a large portion of G was underglycosylated, resulting in the addition of either one or no oligosaccharide to G. Addition of a single oligosaccharide occurred randomly rather than preferentially at one of the two sites. PMID- 9328826 TI - Detection and cloning of epidermal zinc-alpha 2-glycoprotein cDNA and expression in normal human skin and in tumors. AB - Zinc-alpha 2-glycoprotein (Zn alpha 2gp) is almost ubiquitous in body fluids, and its antibody labels the corresponding secretory epithelia. We have found that Zn alpha 2gp is also expressed in human epidermis. We cloned the Zn alpha 2gp cDNA by screening our cDNA library, derived from epidermal keratinocytes, with a probe for prostate Zn alpha 2gp. It had complete nucleic acid sequence homology with that from prostate, including the signal peptide. Just as Zn alpha 2gp expression is higher in more differentiated breast tumors, so in skin tumors the highest mRNA levels occurred in the normal controls, the lowest in basal cell carcinomas (the least differentiated epidermal tumor type), and intermediate levels in squamous cell carcinomas and Merkel cell carcinomas. A similar increase in Zn alpha 2gp gene expression with differentiation was observed when epidermal keratinocytes were cultured in media that varied in cellular maturation potential. PMID- 9328827 TI - Role of cell cycle regulators in tumor formation in transgenic mice expressing the human neurotropic virus, JCV, early protein. AB - Transgenic mice harboring the early genome from the human neurotropic JC virus, JCV, develop massive abdominal tumors of neural crest origin during 6-8 months after birth and succumb to death a few weeks later. The viral early protein, T antigen, which possesses the ability to transform cells of neural origin, is highly expressed in the tumor cells. Immunoblot analysis of protein extract from tumor tissue shows high level expression of the tumor suppressor protein, p53, in complex with T-antigen. Expression of p21, a downstream target for p53, which controls cell cycle progression by regulating the activity of cyclins and their associated kinases during the G1 phase, is extremely low in the tumor cells. Whereas the level of expression and activity of cyclin D1 and its associated kinase, cdk6, was modest in tumor cells, both cyclin A and E, and their kinase partners, cdk2 and cdk4, were highly expressed and exhibited significant kinase activity. The retinoblastoma gene product, pRb, which upon phosphorylation by cyclins:cdk induces rapid cell proliferation, was found in the phosphorylated state in tumor cell extracts, and was detected in association with JCV T-antigen. The transcription factor, E2F-1, which dissociates from the pRb-E2F-1 complex and stimulates S phase-specific genes upon phosphorylation of pRb and/or complexation of pRb with the viral transforming protein, was highly expressed in tumor cells. Accordingly, high level expression of the E2F-1-responsive gene, proliferating cell nuclear antigen (PCNA), was detected in the tumor cells. These observations suggest a potential regulating pathway that, upon expression of JCV T-antigen, induces formation and progression of tumors of neural origin in a whole animal system. PMID- 9328828 TI - Apoptosis in C3H-10T1/2 cells: roles of intracellular pH, protein kinase C, and the Na+/H+ antiporter. AB - Changes in intracellular ion concentrations have been correlated with the activation of an endogenous endonuclease and thus internucleosomal DNA cleavage during apoptosis in many cell types. We investigated whether intracellular pH could play a significant role in apoptotic initiation and progression in C3H 10T1/2 cells, a cell strain that does not exhibit double-stranded DNA cleavage during apoptosis. Protein kinase C and the Na+/H+ antiporter, known regulators of intracellular pH, also were assessed for their involvement in apoptosis of C3H 10T1/2 cells. When a H+ ionophore was used to clamp intracellular pH to 6.0 or below, a significant level of apoptosis was induced in these cells within 6 h, whereas clamping at pH 6.75 did not induce significant amounts of apoptosis until 36 h after acidification. The acidified cells exhibited classic apoptotic morphology and chromatin condensation, similar to serum withdrawn cells, but failed to show internucleosomal DNA cleavage with electrophoresis of genomic DNA. Our results also suggest that the 12-O-tetradecanoylphorbol-13-acetate (TPA) mediated inhibition of apoptosis in serum withdrawn C3H-10T1/2 cells functions through a sequential activation of protein kinase C and the Na+/H+ antiporter; thus, an alkalinization or an inhibition of acidification is involved in this apoptotic block. Serum withdrawal itself does not appear to act through a negative effect on either protein kinase C or the Na+/H+ antiporter. TPA was also capable of inhibiting the apoptosis induced by specific inhibitors of protein kinase C and the Na+/H+ antiporter, but the inhibition was successful only if the TPA was administered at least 20 min prior to the addition of the enzyme inhibitor. These results indicate that apoptosis in C3H-10T1/2 cells follows a pathway that involves intracellular acidification, but is independent of detectable endonuclease activity. PMID- 9328829 TI - In vitro motility assay of atrial and ventricular myosin from pig. AB - The role of myosin isoforms in determining contractile filament velocity in the atrium and ventricle of the pig heart was studied by measuring the motion of fluorescently labeled actin over myosin (in vitro motility assay). A rapid and relatively simple method for purification of myosin from small tissue samples was used. The relative extent of light chain-2 phosphorylation was about 30% in both atrial and ventricular myosin extracts. Although the extracted myosin was not free from contaminating proteins, mainly actin, the mean velocity at optimal pH and 32 degrees C of both atrial (3.3 microns/s) and ventricular (2.3 microns/s) myosin were similar to those obtained using extensively purified myosin. The filament sliding velocities using isolated myosin and actin are lower than those estimated from previously published experiments on skinned fiber preparations, which might reflect an influence on sliding velocity by the filament organization or regulatory proteins in the muscle fiber. However, the ratio between velocities of atrial and ventricular myosin was similar in the motility assay (1.5) and muscle fiber experiments (1.6), which might suggest that these two methods reflect the same fundamental processes in cardiac contraction and that the difference in filament sliding velocity between the atrium and ventricle of the pig heart is determined my their myosin isoforms. PMID- 9328830 TI - Inhibition of osteoblastic cell differentiation by conditioned medium derived from the human prostatic cancer cell line PC-3 in vitro. AB - Human prostatic carcinoma frequently metastasizes to bone tissue and activates bone metabolism, especially bone formation, at the site of metastasis. It has been reported that an extract of prostatic carcinoma and conditioned medium (CM) of a human prostatic carcinoma cell line, PC-3, established from a bone metastastic lesion, stimulate osteoblastic cell proliferation. However, there is little information about the effect of PC-3 CM on the differentiation of osteoblastic cells. In this study, we investigated the effect of PC-3 CM on the differentiation of two types of osteoblastic cells, primary fetal rat calvaria (RC) cells containing many undifferentiated osteoprogenitor cells, and ROS 17/2.8, a well-differentiated rat osteosarcoma cell line. PC-3 CM inhibited bone nodule formation and the activity of alkaline phosphatase (ALPase), an osteoblastic marker enzyme, on days 7, 14, and 21 (RC cells) or 3, 6, and 9 (ROS 17/2.8 cells) in a dose-dependent manner (5-30% CM). However, the CM did not affect cell proliferation or cell viability. PC-3 CM was found to markedly block the gene expression of ALPase and osteocalcin (OCN) mRNAs but had no effect on the mRNA expression of osteopontin (OPN), the latter two being noncollagenous proteins related to bone matrix mineralization. These findings suggest that PC-3 CM contains a factor that inhibits osteoblastic cell differentiation and that this factor may be involved in the process of bone metastasis from prostatic carcinoma. PMID- 9328832 TI - Parathyroid hormone (1-34)-mediated interleukin-6 induction. AB - Parathyroid hormone (PTH) functions in part by regulating osteoblast cytokine expression. We recently demonstrated that PTH induced a rapid and transient increase in interleukin-6 (IL-6) mRNA expression in rat bones in vivo. To determine the molecular basis of this effect, we analyzed the human IL-6 promoter fused (-1,179 to +9) with the chloramphenicol acetyltransferase (CAT) reporter gene in stable transfections into human osteoblast-like osteosarcoma SaOS-2 cells. We compared the effects of PTH on IL-6 expression with adenylate cyclase activator forskolin, PKC activator phorbol 12-myristate 13-acetate (PMA), calcium ionophore A23187, interleukin-1 alpha (IL-1 alpha), prostaglandin E-2 (PGE-2), RS 66271 (a parathyroid hormone-related peptide analog), and platelet-derived growth factor-BB (PDGF-BB). Analyses of cell clones showed that IL-6 promoter expression was extremely low in the unstimulated state. Exposure to PTH (0.001-100 nM) for 12 h stimulated CAT expression in a dose-dependent manner (200-500% of control). Treatment with IL-1 alpha was more potent than PTH in inducing transcription of the IL-6 promoter (900-1,000%). Activation of the cAMP-PKA pathway by treatment with forskolin induced a comparable level of induction with PTH. Together, the effects of PTH and forskolin were additive. RS-66271, previously shown to have PTH-like effects, induced a comparable level of IL-6 promoter expression. When examined together, PTH+RS-66271 effects were comparable to PTH effects alone. Exposure to PGE-2, PMA, PDGF-BB, or A23187 for 12 h did not significantly alter IL-6 promoter expression. These results demonstrate PTH, forskolin, the PTHrP analog RS-66271, and IL-1 alpha stimulate IL-6 expression by stimulating gene transcription. The response to forskolin suggests that the messenger system mediated by PKA is sufficient to induce IL-6 expression. PMID- 9328831 TI - Purification and characterization of a silica-induced bronchoalveolar lavage protein with fibroblast growth-promoting activity. AB - Experimentally induced silicosis provides a good model for chronic interstitial pulmonary inflammation and fibrosis. In the present study, a specific single polypeptide with an apparent molecular mass of 58,000 and a pl of 4.5 was purified and characterized from the bronchoalveolar lavage fluid of silicotic rats. The same protein was also isolated from both the extract and conditioned medium of alveolar macrophages of silicotic rats. Therefore, this protein was termed an inducible silicotic (rat) bronchoalveolar lavage protein-p58 (iSBLP58) or an inducible silicotic (rat) pulmonary macrophage factor (iSPMF-p58). iSBLP58 has been purified to homogeneity by a combination of gel permeation, Mono Q ion exchange, and reverse-phase high performance liquid chromatography. This polypeptide displayed a potent fibroblast growth-promoting activity in vitro. The sequence of the first 15 NH2-terminal amino acids was determined and was found to have high sequence homology with members of the mammalian chitinase-like protein family, which includes human cartilage gp39, mammalian oviduct-specific glycoprotein, and a secretory protein from activated mouse macrophages. PMID- 9328833 TI - Novel nuclear matrix protein HET binds to and influences activity of the HSP27 promoter in human breast cancer cells. AB - Since the small heat shock protein hsp27 enhances both growth and drug resistance in breast cancer cells, and is a bad prognostic factor in certain subsets of breast cancer patients, we have characterized the transcriptional regulation of hsp27, with the long-term goal of targeting its expression clinically. The majority of the promoter activity resides in the most proximal 200 bp. This region contains an imperfect estrogen response element (ERE) that is separated by a 13-bp spacer that contains a TATA box. Gel-shift analysis revealed the binding of a protein (termed HET for Hsp27-ERE-TATA-binding protein) to this region that was neither the estrogen receptor nor TATA-binding protein. We cloned a complete cDNA (2.9 kb) for HET from an MCF-7 cDNA library. To confirm the identity of the HET clone, we expressed a partial HET clone as a glutathione S-transferase fusion protein, and showed binding to the hsp27 promoter fragment in gel-retardation assays. The HET clone is almost identical to a recently published scaffold attachment factor (SAF-B) cloned from a HeLa cell cDNA library. Scaffold attachment factors are a subset of nuclear matrix proteins (NMP) that interact with matrix attachment regions. Analyzing how HET could act as a regulator of hsp27 transcription and as a SAF/NMP, we studied its subnuclear localization and its effect on hsp27 transcription in human breast cancer cells. We were able to show that HET is localized in the nuclear matrix in various breast cancer cell lines. Furthermore, in transient transfection assays using hsp27 promoter luciferase reporter constructs, HET overexpression resulted in a dose-dependent decrease of hsp27 promoter activity in several cell lines. PMID- 9328834 TI - Embryonic morphogenesis signaling pathway mediated by JNK targets the transcription factor JUN and the TGF-beta homologue decapentaplegic. AB - The dorsal surface of the Drosophila embryo is formed by the migration of the lateral epithelial cells to cover the amnioserosa. The Drosophila cJun-N-terminal kinase (DJNK) is essential for this process. Mutations in DJNK or the DJNK activator hemipterous (HEP) lead to incomplete dorsal closure, resulting in a hole in the dorsal cuticle. The molecules downstream of DJNK in this signaling pathway have not been established. Here we demonstrate that the basket1 (bsk1) mutation of DJNK causes decreased interaction with DJUN. Expression of decapentaplegic (DPP), a TGF-beta homologue, in the leading edge of the dorsal epithelium, is identified as a genetic target of the JNK pathway. A constitutive allele of JUN is able to rescue the dorsal closure defect of bsk1 and restores DPP expression. Furthermore, ectopic DPP rescues the defects in dorsal closure caused by bsk1. These data indicate that the interaction of DJNK with DJUN contributes to the dorsal closure signaling pathway and targets DPP expression. PMID- 9328835 TI - Downregulation of telomerase activity in HL60 cells by differentiating agents is accompanied by increased expression of telomerase-associated protein. AB - Telomerase activity provides a mechanism for the unlimited division potential of neoplastic cells. Induced differentiation of these cells was found to be associated with repression of telomerase activity irrespective of the inducing agent. We have employed a series of sublines of human promyelocytic leukemia line HL60 with differing degrees of resistance to differentiation to determine how tightly the expression of the differentiated phenotype is coupled to the downregulation of telomerase activity and to the expression of the recently identified telomerase-associated protein 1 (TP1). As expected, in the 1,25D3 dihydroxyvitamin D3 (1,25D3)-resistant subclones (20A-100A cells), telomerase activity was not significantly downregulated by 1,25D3 and, in most cases, by all trans retinoic acid (atRA), to which these cells were cross-resistant, but telomerase activity was repressed by dimethylsulfoxide (DMSO) and phorbol-12 myristate-13-acetate (TPA), to which the sublines were in general sensitive. However, there were exceptions; in some instances telomerase activity was repressed in the absence of the expression of markers of differentiation. Also, there was an inverse relationship between telomerase activity and the cellular levels of TP1 transcripts. We conclude that in HL60 cells downregulation of telomerase is loosely associated with upregulation of differentiation markers and with other cellular changes which include an upregulation of TP1. PMID- 9328836 TI - Distinct roles for Sp1 and E2F sites in the growth/cell cycle regulation of the DHFR promoter. AB - Dihydrofolate reductase activity is required for many biosynthetic pathways including nucleotide synthesis. Its expression is therefore central to cellular growth, and it has become a key target for cancer chemotherapy. Transcription of the dihydrofolate reductase gene is regulated with growth, being expressed maximally in late G1/early S phase following serum stimulation of quiescent cells. This regulation is directed by a promoter which contains binding sites for only the transcription factors Sp1 and E2F. In this study, the role of these promoter elements in growth/cell cycle regulation of dihydrofolate transcription was addressed directly by transient transfection of Balb/c 3T3 cells with mutant promoter-reporter gene constructs. The E2F sites were found to repress transcription in G0 and early G1 but did not contribute to the level of transcription in late G1/S phase. In contrast, Sp1 sites were able to mediate induction of transcription from the dihydrofolate reductase promoter, as well as a heterologous promoter, following serum stimulation of quiescent cells. These findings add dihydrofolate reductase to a growing list of genes at which E2F sites are primarily repressive elements and delineate a role for Sp1 sites in the growth/cell cycle regulation of transcription. PMID- 9328837 TI - Activation-induced bi-dentate interaction of SHIP and Shc in B lymphocytes. AB - SHIP is a SH2 domain-containing inositol polyphosphatase that is selectively tyrosine phosphorylated and associated with the adapter protein Shc in B lymphocytes upon co-crosslinking surface immunoglobulin and Fc gamma RIIB1. We previously observed that this stimulation condition is associated with a reduction in the interaction of Grb2 with phosphorylated Shc, an enhanced interaction of Shc with SHIP, and a block in the Ras signaling pathway. We proposed that the SH2 domain of SHIP competes with Grb2 in binding to phospho Shc, resulting in a block in Ras signaling. To test this model, we examined the mode of SHIP-Shc interaction. Using recombinant Shc and SHIP interaction domains and purified Shc and SHIP phosphopeptides, we show that the interaction is bi dentate such that the SH2 domain of SHIP recognizes phosphorylated Y317 and doubly-phosphorylated Y239/Y240 of Shc and the Shc PTB domain recognizes phosphorylated NPxpY motifs within SHIP. We observed no role for the Shc SH2 domain in the interaction. These findings are consistent with our earlier model that SHIP and Grb2 compete for binding to phospho-Shc and support the notion that, in addition to the hydrolysis of inositol phosphates and phospholipids, SHIP contributes to anti-proliferative biochemistry by blocking protein-protein interactions. PMID- 9328839 TI - Suppression of extracellular signals and cell proliferation through EGF receptor binding by (-)-epigallocatechin gallate in human A431 epidermoid carcinoma cells. AB - Tea polyphenols are known to inhibit a wide variety of enzymatic activities associated with cell proliferation and tumor progression. The molecular mechanisms of antiproliferation are remained to be elucidated. In this study, we investigated the effects of the major tea polyphenol (-)-epigallocatechin gallate (EGCG) on the proliferation of human epidermoid carcinoma cell line, A431. Using a [3H]thymidine incorporation assay, EGCG could significantly inhibit the DNA synthesis of A431 cells. In vitro assay, EGCG strongly inhibited the protein tyrosine kinase (PTK) activities of EGF-R, PDGF-R, and FGF-R, and exhibited an IC50 value of 0.5-1 microgram/ml. But EGCG scarcely inhibited the protein kinase activities of pp60v-src, PKC, and PKA (IC50 > 10 micrograms/ml). In an in vivo assay, EGCG could reduce the autophosphorylation level of EGF-R by EGF. Phosphoamino acid analysis of the EGF-R revealed that EGCG inhibited the EGF stimulated increase in phosphotyrosine level in A431 cells. In addition, we showed that EGCG blocked EGF binding to its receptor. The results of further studies suggested that the inhibition of proliferation and suppression of the EGF signaling by EGCG might mainly mediate dose-dependent blocking of ligand binding to its receptor, and subsequently through inhibition of EGF-R kinase activity. PMID- 9328838 TI - Analysis of the phosphorylation of human heat shock transcription factor-1 by MAP kinase family members. AB - The activation of heat shock transcription factor-1 (HSF-1) after treatment of mammalian cells with stresses such as heat shock, heavy metals, or ethanol induces the synthesis of heat shock proteins. HSF-1 is phosphorylated at normal growth temperature and is hyperphosphorylated upon stress. We recently presented evidence that HSF-1 can be phosphorylated by the mitogen activated protein kinase, ERK1, and that such phosphorylation appears to negatively regulate the activity of HSF-1. In this report, we have tested the ability of ERK1 to phosphorylate various HSF-1 deletion mutants. Our results show that ERK1 phosphorylation is dependent on a region of HSF-1 extending from amino acids 280 to 308. This region contains three serine residues that are potential ERK1 phosphorylation sites. The region falls within a previously defined regulatory domain of HSF-1. The possibility of protein kinases other than ERK1 phosphorylating HSF-1 was also examined using in-gel kinase assays. The results show that HSF-1 can be phosphorylated in a ras-dependent manner by other members of the MAP kinase family such as JNK and p38 protein kinases and possibly others. PMID- 9328840 TI - Scleraxis messenger ribonucleic acid is expressed in C2C12 myoblasts and its level is down-regulated by bone morphogenetic protein-2 (BMP2). AB - We examined the mRNA expression of scleraxis, a non-myogenic helix-loop-helix type transcription factor in C2C12 myogenic cells. Scleraxis mRNA has been shown to be expressed in sclerotome and perichondrium of the embryos. We found that C2C12 cells express 1.2 kb scleraxis mRNA constitutively. Since BMP was reported to induce ectopic bone formation when implanted in muscle, we examined the effects of BMP on scleraxis expression. Scleraxis mRNA expression in C2C12 cells was suppressed by the treatment with BMP2. This suppression was observed at 200 ng/ml but not at the lower concentrations. BMP2 treatment suppressed scleraxis mRNA level within 24 h and lasted at least up to 48 h. Electrophoresis mobility shift assay showed that the proteins in the crude nuclear extracts prepared from C2C12 cells bound to an Scx-E-box sequence, CATGTG, which is preferentially recognized by scleraxis. This binding was competed out by 100-fold molar excess of cold Scx-E-box sequence but not by the one with mutations in the E-box. This band was supershifted by the addition of antiserum raised against scleraxis. BMP2 treatment suppressed the Scx-E binding activity in C2C12 cells. This suppression of the Scx-E-box binding activity was in parallel to the BMP2 suppression of the transcriptional activity of the Scx-E-CAT reporter gene transfected into C2C12 cells. These data indicated that although the default pathway for C2C12 cells is to differentiate into muscle cells, these cells do express non-myogenic transcription factor, scleraxis, whose expression is suppressed by BMP2. PMID- 9328841 TI - Decorin inhibits cell attachment to thrombospondin-1 by binding to a KKTR dependent cell adhesive site present within the N-terminal domain of thrombospondin-1. AB - Skin decorin (DCN) is an antiadhesive dermatan sulfate-rich proteoglycan that interacts with thrombospondin-1 (TSP) and inhibits fibroblast adhesion to TSP [Winnemoller et al., 1992]. Molecular mechanisms by which DCN interacts with TSP and inhibits cell adhesion to TSP are unknown. In the present study, we showed that skin DCN and bone DCN (chondroitin sulfate-rich proteoglycan) were quantitatively identical with respect to their ability to interact with TSP. Using a series of fusion proteins corresponding to the different structural domains of TSP, binding of [125I]DCN to TSP was found to be dependent of the N terminal domain and, to a lesser extent, of the type 1 repeats and the C-terminal domain of TSP. In addition, heparan sulfate drastically inhibited [125I]DCN binding to solid-phase adsorbed TSP (80% inhibition), suggesting that DCN could bind to the N-terminal domain of TSP through interaction with heparin-binding sequences. To address this question, a series of synthetic peptides, overlapping heparin-binding sequences ARKGSGRR (residues 22-29), KKTR (residues 80-83) and RLRIAKGGVNDN (residues 178-189), were synthesized and tested for their ability to interact with DCN. [125I]DCN interacted only with peptides VDAVRTEKGFLLLASLRQMKKTRGT and KKTRGTLLALERKDHS containing the heparin-binding consensus sequence KKTR. These peptides contained glycosaminoglycan-dependent and -independent binding sites because [125I]DCN binding to VDAVRTEKGFLLLASLRQMKKTRGT and KKTRGTLLALERKDHS was partially reduced upon removal of the glycosaminoglycan chain (65% and 46% inhibition, respectively). [125I]DCN poorly bound to subpeptide MKKTRG and did not bind at all to subpeptides VDAVRTEKGFLLLASLRQ and TLLALERKDHS, suggesting that heparin-binding sequence MKKTRG constituted a DCN binding site when flanked with peptides VDAVRTEKGFLLLASLRQ and TLLALERKDHS. The sequence VDAVRTEKGFLLLASLRQMKKTRGTLLALERKDHS constitutes a cell adhesive active site in the N-terminal domain of TSP [Clezardin et al., 1997], and DCN inhibited the attachment of fibroblastic and osteoblastic cells to peptides VDAVRTEKGFLLLASLRQMKKTRGT and KKTRGTLLALERKDHS by about 50 and 80%, respectively. Although fibroblastic cells also attached to type 3 repeats and the C-terminal domain of TSP, DCN only inhibited cell attachment to the C-terminal domain. Overall, these data indicate that modulation by steric exclusion of cell adhesion to a KKTR-dependent cell adhesive site present within the N-terminal domain of TSP could explain the antiadhesive properties of DCN. PMID- 9328842 TI - Reorganization of a novel vimentin-associated protein in 3T3-L1 cells during adipose conversion. AB - We have found that the antibody A2, a marker for the capsule of steroidogenic lipid droplets, reacts with an intermediate filament-associated protein, P200, in 3T3-L1 preadipocytes. Supporting evidence came from the colocalization pattern of P200 with vimentin in double label experiments. The association of P200 with vimentin was further confirmed by its copurification with vimentin after high salt extraction and colocalization of these two proteins in high salt-extracted and vinblastine-treated cells. In preadipocytes this protein was distributed on the vimentin filament network. At the early stage of adipose conversion, this protein was found to encircle nascent lipid droplets ranging from 0.1 to 0.2 micron, accompanied with a decreased distribution on the vimentin filament system. This infers a possible translocation of P200 from the vimentin filaments to the droplet surface. Meanwhile, the vimentin filaments remained in a normal distribution in the cytoplasm and were apparently not associated with the nascent droplet. The association of vimentin filaments to droplet surfaces became prominent in lipid droplets larger than 0.2 micron, forming a typical vimentin cage. Immunogold staining also confirmed the translocation of P200 immunoreactivity from the droplet surface to the vimentin cage. The relocation of P200 from the cytoplasmic vimentin filaments to the droplet surface prior to the formation of the vimentin cage, as well as the reorganization of this protein in the vimentin cage, suggests a stabilizing role in the lipid droplet formation and an inducing function of this protein in the formation of the vimentin cage. PMID- 9328843 TI - Regulation of the rat interstitial collagenase promoter by IL-1 beta, c-Jun, and Ras-dependent signaling in growth plate chondrocytes. AB - In an attempt to better define molecular influences on rat interstitial collagenase gene expression in cartilage, the promoter function was characterized using transient transfection assay, electrophoresis mobility shift assay, and genetic analysis in isolated growth plate chondrocytes. Data from 5'-flanking deletion and selected mutations suggest that multiple cis elements in both the proximal and distal regions of the promoter were important in the regulation of promoter activity. A proximal tumor response element (TRE) was shown to be necessary for basal and interleukin (IL)-1 beta-inducible reporter gene activity. Cells stimulated by IL-1 beta (1 ng/ml; 18 h) had elevated TRE binding activity, and one of the factors involved was identified as the nuclear protein, c-Jun. Indeed, c-Jun directed antisense oligonucleotides reduced rat interstitial collagenase mRNA. A sense oligonucleotide was ineffective. Regulation of promoter activity was susceptible to Ras-dependent signaling as expression of dominant negative mutant of Ras kinase (pZIP-RasN17) reduced reporter gene activity. In a comparison of proximal promoter reporter plasmid activity between proliferative and hypertrophic cells, inhibition of Ras-dependent signaling was less effective in the later cell type. This study suggests that the activation of nuclear binding proteins that bind TRE may be a common event with IL-1 beta regulation. Moreover, these data suggest that the regulation of rat interstitial collagenase gene expression is a combinatorial process and multiple cis-acting regulatory sites may interact to exert different effects dependent on the stage of chondrocyte differentiation. PMID- 9328844 TI - Protein kinase C activation by interleukin (IL)-1 limits IL-1-induced IL-6 synthesis in osteoblast-like cells: involvement of phosphatidylcholine-specific phospholipase C. AB - We investigated the regulatory mechanism of interleukin-6 (IL-6) synthesis induced by interleukin-1 (IL-1) in osteoblast-like MC3T3-E1 cells. IL-1 stimulated the secretion of IL-6 in a dose-dependent manner in the range between 0.1 and 100 ng/ml. Staurosporine and calphostin C, inhibitors of protein kinase C (PKC), significantly enhanced the IL-1-induced secretion of IL-6. The stimulative effect of IL-1 was markedly amplified in PKC down-regulated MC3T3-E1 cells. IL-1 produced diacylglycerol in MC3T3-E1 cells. IL-1 had little effect on the formation of inositol phosphates and choline. On the contrary, IL-1 significantly stimulated the formation of phosphocholine dose-dependently. D-609, an inhibitor of phosphatidylcholine-specific phospholipase C, suppressed the IL-1-induced diacylglycerol production. The IL-1-induced IL-6 secretion was significantly enhanced by D-609. These results indicate that IL-1 activates PKC via phosphatidylcholine-specific phospholipase C in osteoblast-like cells, and the PKC activation then limits IL-6 synthesis induced by IL-1 itself. PMID- 9328845 TI - Quantitative immunofluorescence and immunoelectron microscopy of the topoisomerase II alpha associated with nuclear matrices from wild-type and drug resistant chinese hamster ovary cell lines. AB - Topo II alpha is considered an important constituent of the nuclear matrix, serving as a fastener of DNA loops to the underlying filamentous scaffolding network. To further define a mechanism of drug resistance to topo II poisons, we studied the quantity of topo II alpha associated with the nuclear matrix in drug resistant SMR16 and parental cells in the presence and absence of VP-16. Nuclear matrices were prepared from nuclei isolated in EDTA buffer, followed by nuclease digestion with DNase II in the absence of RNase treatment and extraction with 2 M NaCl. Whole-mount spreading of residual structures permits, by means of isoform specific antibody and colloidal-gold secondary antibodies, an estimate of the amount of topo II alpha in individual nuclear matrices. There are significant variations in topo II alpha amounts between individual nuclear matrices due to the cell cycle distribution. The parental cell line contained eight to ten times more nuclear matrix-associated topo II alpha than the resistant cell line matrices. Nuclear matrix-associated topo II alpha from wild-type and resistant cell lines correlated well with the immunofluorescent staining of the enzyme in nuclei of intact cells. The amount of DNA associated with residual nuclear structures was five times greater in the resistant cell line. This quantity of DNA was not proportional to the quantity of topo II alpha in the same matrix; in fact they were inversely related. In situ whole-mount nuclear matrix preparations were obtained from cells grown on grids and confirmed the results from labeling of isolated residual structures. PMID- 9328846 TI - Increases in mRNA levels of glucose transporters types 1 and 3 in Ehrlich ascites tumor cells during tumor development. AB - A common feature of many tumors is an increase in glucose catabolism during tumor growth. We studied the mechanism of this phenomenon by using Ehrlich ascites tumor bearing mice as the animal model. We found that Ehrlich ascites tumor cells possess only glucose transporter 1 (GLUT1) and GLUT3 but not GLUT2, GLUT4, or GLUT5. The mRNA levels of GLUT1 and GLUT3 increased progressively in the tumour during development; however, there were no changes observable in mRNA levels of glucose transporters of all types in brain, liver, and heart of the host mice. These findings suggest that Ehrlich ascites tumor augments its glucose transport mechanism relative to other tissues in response to its unique growth needs. PMID- 9328847 TI - Expression of meltrin-alpha mRNA is not restricted to fusagenic cells. AB - Meltrin-alpha is a myoblast gene product reported to be required for cell fusion [Yagami-Hiromasa et al. (1995): Nature 377:652-656]. Because Northern blots revealed expression only in muscle and bone, the suggestion was made that meltrin alpha is expressed exclusively by fusagenic cells in these tissues (myoblast and osteoclast). We studied expression of meltrin-alpha mRNA in a panel of tissues and cell lines using the polymerase chain reaction and found it widely expressed. Meltrin-alpha mRNA was readily detected in the osteoblast, the most abundant cell type in bone. In situ hybridization analysis on sections of neonatal mice revealed high levels of expression in the trabecular meshwork of long bones, the basal regions of the dermis and its underlying mesenchyme. We conclude that expression of meltrin-alpha mRNA is not restricted to fusagenic cells and that, in bone, the osteoblast is the major source. PMID- 9328848 TI - Transgene-coded chimeric proteins as reporters of intracellular proteolysis: starvation-induced catabolism of a lacZ fusion protein in muscle cells of Caenorhabditis elegans. AB - The product of an integrated transgene provides a convenient and cell-specific reporter of intracellular protein catabolism in 103 muscle cells of the nematode Caenorhabditis elegans. The transgene is an in-frame fusion of a 5'-region of the C. elegans unc-54 (muscle myosin heavy-chain) gene to the lacZ gene of Escherichia coli [Fire and Waterston (1989): EMBO J 8:3419-3428], encoding a 146 kDa fusion polypeptide that forms active beta-galactosidase tetramers. The protein is stable in vivo in well-fed animals, but upon removal of the food source it is inactivated exponentially (t1/2 = 17 h) following an initial lag of 8 h. The same rate constant (but no lag) is observed in animals starved in the presence of cycloheximide, implying that inactivation is catalyzed by pre existing proteases. Both the 146-kDa fusion polypeptide (t1/2 = 13 h) and a major 116-kDa intermediate (t1/2 = 7 h) undergo exponential physical degradation after a lag of 8 h. Degradation is thus paradoxically faster than inactivation, and a number of characteristic immunoreactive degradation intermediates, some less than one-third the size of the parent polypeptide, are found in affinity-purified (active) protein. Some of these intermediates are conjugated to ubiquitin. We infer that the initial proteolytic cleavages occur in the cytosol, possibly by a ubiquitin-mediated proteolytic pathway and do not necessarily inactivate the fusion protein tetramer. PMID- 9328849 TI - Changes in thoracopulmonary compliance and hemodynamic effects of positive end expiratory pressure in patients with or without heart failure. AB - PURPOSE: The purpose of this study was to confirm that positive end-expiratory pressure (PEEP) has a different effect on cardiac index (CI) in patients with or without heart failure, even after controlling for differences in thoracopulmonary compliance (Ctp) and minimizing the secondary effects of PEEP related changes in oxygenation and breathing effort. MATERIALS AND METHODS: The hemodynamic effects of PEEP were evaluated in two groups of sedated and paralyzed patients with a low Ctp at 0 PEEP: 12 patients with normal pulmonary artery occlusion pressure (Ppao) and a CI > 2.5 L/min and 12 patients with a CI < 2.5 L/min and increased oxygen extraction ratio, despite a Ppao > 15 mm Hg. RESULTS: In patients with low CI and high Ppao, PEEP had no hemodynamic effect and Ctp remained low at all PEEP levels. However, PEEP-induced CI reduction in patients with normal cardiovascular function was associated with an increase in Ctp with incremental PEEP. Concerning PEEP-related hemodynamic effects, the significance between group differences persisted when data were analyzed after controlling for Ctp changes. However, Ctp changes with PEEP were the most significant correlators and discriminators of the magnitude and direction of PEEP-induced CI change. CONCLUSIONS: We conclude that (1) the observed different effect of PEEP on CI in patients with and without heart failure persists after the elimination of secondary effects due to underlying differences in Ctp, oxygenation, and breathing effort; and (2) PEEP related changes in Ctp should be taken into consideration when dealing with the cardiovascular effects of PEEP. Our data support the hypothesis that, in addition to the transmission of PEEP to the pleural space, changes in lung volume are a significant determinant of PEEP-induced CI changes. PMID- 9328850 TI - Continuous jugular bulb venous oxygen saturation validation and variations during intracranial aneurysm surgery. AB - PURPOSE: During intracranial aneurysm surgery, numerous factors may alter cerebral blood flow and oxygen supply-demand balance. Continuous monitoring of jugular bulb venous oxygen saturation (SvjO2) may help in the anesthetic management of such procedures. MATERIALS AND METHODS: Fiberoptic SvjO2 was continuously monitored in seven patients during intracranial aneurysm surgery. Fiberoptic SvjO2 measurement was compared with IL3 CO-OXIMETER determination from 85 paired samples. The occurrence of large SvjO2 variations (SvjO2 variation reaching 10% or more of stable preceding value) during aneurysm surgery was recorded and classified according to the association or not with systemic clinical or therapeutic changes. RESULTS: Fiberoptic SvjO2 showed a limited accuracy, with limits of agreement with IL3 CO-OXIMETER at -16.8% and +10.7% and a small bias (-3.1%). SvjO2 variations were frequent during aneurysm surgery, ranging from 3 to 22 per patient during procedures lasting 6 hours (range 4.5 to 7). Half of these variations occurred in the absence of any systemic clinical or therapeutic change, most often leading to an increased SvjO2. CONCLUSIONS: Although the accuracy of fiberoptic SvjO2 determination is limited, it allows the detection of cerebral blood flow and oxygen supply-demand imbalance during aneurysm surgery. The frequent occurrence of SvjO2 elevations is suggestive of reactive hyperemia mechanisms. PMID- 9328852 TI - The effects of intravenous anesthetics on intracranial pressure and cerebral perfusion pressure in two feline models of brain edema. AB - PURPOSE: The purpose of this study was to investigate the effects of various intravenous anesthetics on intracranial pressure (ICP) and cerebral perfusion pressure (CPP) in two models of brain edema in a prospective study in a Pediatric critical care animal laboratory in a university hospital. MATERIALS AND METHODS: Intraparenchymal ICP monitors were inserted in 30 anesthetized adult cats. In 15 cats, an intraparenchymal balloon-tipped catheter was placed and inflated to create a space-occupying lesion (SOL) to mimic vasogenic brain edema (VBE). In the other 15 cats, cytotoxic brain edema (CBE) was created by an acute reduction in blood osmolality. We used continuous hemodiafiltration (CAVH-D) and replaced the ultrafiltrate with hypotonic solution to maintain euvolemia. At predetermined points, each cat in each model received multiple intravenous (i.v.) injections of one of the following medications: methohexital 1.5 mg/kg, propofol 2 mg/kg, or ketamine 2 mg/kg. ICP and mean arterial pressure (MAP) were continuously monitored in all animals. RESULTS: In the SOL model, all three anesthetic agents decreased ICP after each administration (P < .05). Ketamine administration also resulted in an increase in CPP in this model (P < .05). In the CBE model, none of these agents resulted in a significant change in either ICP or CPP. CONCLUSIONS: Our results indicate that i.v. anesthetics decrease ICP caused by SOL but have no significant effect on ICP due to CBE. We postulate that in the SOL model, and similarly in VBE, some brain tissue is viable and remains responsive to anesthetics. In contrast, in the CBE model, diffuse intracellular damage occurs, the cerebral metabolic rate may be severely depressed, autoregulation of the cerebral vasculature may be impaired, and unresponsiveness to i.v. anesthetics may occur. PMID- 9328851 TI - Influence of fluid resuscitation on renal function in bacteremic and endotoxemic rats. AB - PURPOSE: Fluid resuscitation, which is the most important primary therapy in sepsis, is not always able to prevent acute renal failure. In this study, we investigated in two different rat models of distributive shock whether fluid resuscitation would increase renal plasma flow (RPF) and subsequently glomerular filtration rate (GFR). MATERIALS AND METHODS: In pentobarbital anesthetized wistar rats Haemaccel (Behring Pharma, Hoechst, the Netherlands) infusion (1.2 mL/100 g/h for 3 hours) was started immediately during either bacteremia (bolus of living Escherichia coli bacteria, 10(9) or endotoxemia (1 hour infusion of E. coli endotoxin, 8 mg/kg), as well as in time-matched healthy controls. RESULTS: After 3 hours, this treatment had increased RPF (clearance of 131I-hippurate) above normal in control (+67%) and bacteremic rats (+75%), whereas in endotoxemic animals, the significantly decreased RPF was normalized. On the other hand, in bacteremic animals, the lowered GFR (clearance of creatinine; x44%) was normalized, whereas in endotoxemic animals GFR remained depressed (x30%). The lack of improvement in GFR during endotoxemia was also indicated by a profound fall in urine flow, which by contrast steadily increased in control and bacteremic rats owing to volume loading. In both shocked groups, the decreased renal oxygen delivery was normalized, but the higher renal oxygen consumption than expected on the basis of the work needed for sodium reabsorption was not influenced by Haemaccel treatment, despite the fact that it caused this work load to rise in bacteremic but not in endotoxemic rats. In both shock models, renal cortical adenosine triphosphate content did not differ from healthy controls and was not influenced by volume loading. CONCLUSIONS: In conclusion, our study suggests that a decrease in GFR caused by live bacteria in the circulation may benefit from fluid resuscitation, while during endotoxemia this therapy could not prevent acute renal failure. PMID- 9328853 TI - Dobutamine maintains intestinal villus blood flow during normotensive endotoxemia: an intravital microscopic study in the rat. AB - PURPOSE: The gut plays a pivotal role in sepsis. Intestinal hypoperfusion with subsequent ischemia leads to translocation of endotoxin. Dobutamine has been demonstrated to increase mesenteric blood flow during endotoxic shock; however, its effects on mucosal blood flow especially in intestinal villi is not known. Therefore, we investigated its influence on the blood flow and the arteriolar diameters in intestinal villi in a model of normotensive endotoxemia. MATERIALS AND METHODS: Twenty-one male Wistar rats were divided into three groups: (1) control, saline; (2) endotoxin, endotoxin 1.5 mg/kg during 60 minutes; and (3) dobutamine, endotoxin 1.5 mg/kg (60 minutes) and dobutamine 2.5 micrograms/kg/min during 120 minutes. Villus blood flow and arteriolar diameters were determined at 0 minutes, 60 minutes, and 120 minutes in each group using intravital microscopy. RESULTS: Villus blood flow was constant in the control group, significantly reduced at 120 minutes in the endotoxin group (120 minutes, 55.1 +/- 7.4%), and remained at baseline values in the dobutamine group. The arteriolar diameters remained constant in the control and the dobutamine groups, but they were significantly reduced in the endotoxin group at 120 minutes (7.8 +/- 0.2 to 6.5 +/- 0.7 micron). CONCLUSION: Our results indicate that in rats with normotensive endotoxemia, arteriolar diameters and blood flow in intestinal villi were reduced. Dobutamine prevented arteriolar constriction and maintained villus blood flow at preendotoxemic values. PMID- 9328854 TI - An outcomes analysis of in-hospital cardiopulmonary resuscitation: the futility rationale for do not resuscitate orders. AB - PURPOSE: Cardiopulmonary resuscitation (CPR) is a frequently performed medical intervention in hospitalized patients who die. Despite the widespread use of do not-resuscitate (DNR) orders during the last decade, the outcome following CPR appears not to have improved. The key to an improved outcome may be better patient selection. The objective of this study was to determine the hospital survival rate following CPR in the era of DNR orders, and to identify risk factors predictive of hospital survival at a university-affiliated teaching hospital. MATERIALS AND METHODS: We retrospectively reviewed the code sheets and patient charts of all patients who underwent CPR during a 4-year period from January 1991 to January 1995. Three-hundred-and-eight patients were identified. RESULTS: CPR was successful in 99 (32%) patients, with 41 (13%) patients surviving to hospital discharge. All the patients who survived were otherwise "healthy" with reversible conditions, who experienced a sudden and unexpected arrhythmic event. No pre-arrest risk factors could clearly distinguish the hospital survivors from the nonsurvivors. The length of the code was 9.4 +/- 4 minutes in the hospital survivors compared with 26.6 +/- 19.1 minutes in the nonsurvivors. Patients whose initial rhythm was either ventricular tachycardia or fibrillation had a better survival rate than patients with other rhythms. CONCLUSION: DNR protocols do not prevent CPR being performed on patients who are unlikely to survive to hospital discharge. CPR should only be offered to patients who are likely to derive benefit from this intervention. PMID- 9328855 TI - Economics and the intensive care unit: a conflict of interests? PMID- 9328856 TI - Central venous pressure, pulmonary artery occlusion pressure, intrathoracic blood volume, and right ventricular end-diastolic volume as indicators of cardiac preload. PMID- 9328857 TI - Harbinger of plague: a bad case of gay bowel syndrome. AB - In 1976, a group of physicians in private proctologic practice in New York City coined the illness "Gay Bowel Syndrome" in reference to a constellation of gay male anorectal disorders. Through analysis of biomedical discourse and popular media, it is apparent that Gay Bowel Syndrome is an essentialized category of difference that is neither gay-specific, confined to the bowel, nor a syndrome. The use and diagnosis of Gay Bowel Syndrome must be abandoned before it further lends itself to the formation of social policies and governing practices that seek to force gay male bodies into positions of social, cultural, and political subordination. PMID- 9328858 TI - The lesbian and gay liberation movement in the Presbyterian Church (U.S.A.), 1974 1996. AB - Since its founding in 1974, Presbyterians for Lesbian & Gay Concerns (PLGC) has led the movement toward full participation and membership for lesbian and gay people in the Presbyterian Church (U.S.A.). This is a story of that movement, told by a participant. It traces the development of current antigay policies in the church in 1976-78 in response to the first openly gay candidate for the ministry, PLGC's efforts to overturn these policies at annual General Assemblies, the growth of the pro-gay More Light Churches movement among Presbyterian congregations, the increasing number of antigay ecclesiastical court cases, study and dialogue on lesbian and gay issues across the denomination, and controversies over same-gender marriage, all culminating in the 1996 General Assembly, which endorsed civil rights for lesbian and gay couples, but also voted to send the controversial issue of lesbian and gay ordination to the 171 regional presbyteries of the church for an up or down vote. The battle for lesbian and gay equity in the church may well continue decades longer. Equity for gays and lesbians in society will not be fully won until the religious establishment learns how to apply its most basic principles of love, nurture, inclusive welcome, and support to lesbian and gay people. PMID- 9328859 TI - A further exploration of the lesbian identity development process and its measurement. AB - This study explored the fit of Cass's (1979) model of homosexual identity formation (HIF) and the utilization of the Self Identity Questionnaire (Brady, 1983) with a sample of lesbian women. Purposes of the study were to evaluate assignment of HIF stage based on two self-report measures, to measure the relationship between aspects of self-concept and lesbian identity development, and to clarify the sequencing of developmental tasks in lesbian identity formation. Participants were 118 women who self-identified as lesbian or as questioning their sexuality. Results of the study indicated that relationships exist between self-reported and assigned HIF stage, that HIF stage correlates with self-concept, and that participants moved through key developmental tasks in the order predicted by Cass's (1979) model. In addition, age was found to have a relationship with HIF stage. The study provides limited support for the utility of the HIF model and its measurement. PMID- 9328860 TI - Should I come out to my students? An empirical investigation. AB - Lesbian, gay, and bisexual educators have faced many barriers in their professions, including harassment, discrimination, and even nationwide antigay political campaigns. Recently, lesbian, gay, and bisexual educators, particularly on college campuses, have challenged such stigmatization by coming out. Because previous research has demonstrated that interpersonal contact with lesbians, gay men, and bisexuals is related to less heterosexist attitudes, the current study investigated the impact of a gay instructor's coming out on his students' attitudes toward lesbians and gay men. Data were collected from 156 undergraduate students enrolled in an Introductory Psychology course, 40 of whom were taught by a gay instructor. Herek's (1984, 1994b) Attitudes Toward Lesbians and Gay Men (ATLG) scale was used to measure students' relative levels of heterosexism and was administered to students at the beginning and end of the semester. Midway through the semester, the gay instructor disclosed his gay identity to his students as part of a lesson about sexual orientation. Results from the postcourse survey indicated that students in the gay instructor's course section exhibited improved attitudes. Conversely, students enrolled in the same course in sections taught by heterosexual instructors demonstrated no change in their attitudes. Implications of these findings are discussed, and it is argued that gay instructors' coming out may positively affect their students' attitudes toward lesbian, gay, and bisexual people. However, these efforts by individual instructors must only be a small part of more comprehensive institutional efforts by university communities to address homophobia and heterosexism in educational settings. PMID- 9328861 TI - Vitamin D3 in Tilapia mossambica: relevance of photochemical synthesis. AB - The capability of fish to synthesize vitamin D on exposure to ultraviolet (UV) light was examined. Purposeful exposure of the freshwater fish Tilapia mossambica (Tilapia) to artificial UV light (300 nm) resulted in a significant increase of vitamin D3 with a concomitant decrease in provitamin D3 [7-dehydrocholesterol (7 DHC)], indicating that provitamin D3 was converted to vitamin D3. However, only 0.13% of the intraperitoneally injected 4-14C cholesterol was recovered in the vitamin D3 and 25-hydroxyvitamin D3 (25-OH-D3) fractions after 15 h of irradiation. Thus, although fish is capable of photosynthesizing vitamin D through constant, prolonged exposure to UV light of appropriate wavelength, the contribution of this mode of synthesis is unlikely to be of any significance in its natural habitat. PMID- 9328862 TI - Effects of antioxidants on the oxidative susceptibility of low-density lipoprotein. AB - An important event in the pathogenesis of atherosclerosis is believed to be the oxidative modification of low-density lipoprotein (LDL) initiated by a free radical-driven lipid peroxidation process. Vitamin E acts as a lipophilic chain breaking antioxidant, while water-soluble chain-breaking antioxidants such as vitamin C or uric acid suppress the oxidation of LDL initiated by aqueous radicals. In this study, we established a new method of measuring the lag time of inhibited lipid peroxidation using the lipophilic azo radical initiator V-70:2-2' azobis(4-methoxy-2,4-dimethylvaleronitrile) and investigated in vitro the susceptibility of LDL to oxidation using this method when lipid- and water soluble antioxidants were added. When the lipid-soluble antioxidant, vitamin E, was added to LDL, the lag time was extended whereas a higher dose of vitamin E led to a shortened lag time of V-70-induced lipid peroxidation in LDL. These results suggest that vitamin E radicals (tocopheroxyl radicals) act as prooxidants during the autoxidation of LDL. It was also shown that the shortened lag time induced by higher doses of vitamin E was restored when lipid- and water soluble antioxidants were added simultaneously, which suggests that vitamin E radicals derived from vitamin E are subsequently reduced by vitamin C to regenerate vitamin E. Thus, the interaction between lipid- and water-soluble antioxidants provides an important function in maintaining LDL resistance to oxidation. PMID- 9328863 TI - Effect of dietary fiber on bowel mucosal integrity and bacterial translocation in burned rats. AB - The response of the bowel mucosa to enteral formula supplemented with dietary fiber was examined in rats with 30% full-thickness burns. The rats were fed a standard enteral formula without fiber or with one of two types of fiber (insoluble soy fiber or soluble guar gum fiber). Seventy-two hours after burn injury, the mesenteric lymph nodes were excised aseptically for bacterial culturing. Samples of the jejunum, ileum and cecum were also collected for histological examination. There were significantly fewer bacterial colonies in the lymph node cultures from rats given soy fiber compared to those from rats fed no fiber. In rats given soy fiber, the integrity of the bowel mucosa was maintained in the jejunum, ileum and cecum. In rats given guar gum fiber, however, the repair of mucosal erosions was observed in the jejunum and ileum as well as flattening of the cecal mucosa. These findings indicate that soy fiber is superior to guar gum fiber for maintaining bowel mucosal integrity and preventing bacterial translocation in burned rats receiving enteral feeding. PMID- 9328864 TI - Anti-tumor activity of squid ink. AB - The anti-tumor activity of a new type of peptidoglycan isolated from squid ink was shown to have a cure rate of 64% for Meth A tumor from BALB/c mice. The ink delipidated in acetone, which contained the peptidoglycan at 0.1% (w/w), was administered to tumor-transplanted mice so as to examine the anti-tumor activity. One-fifth of the tumor-bearing mice was cured with 3 injections (1 mg/head) of the acetone delipidated squid ink or a prolongation of survival was observed in the treated animals. Heat treatment at 100 degrees C for 10 min did not affect the anti-tumor activity of the delipidated ink, its potentiality being preserved. The acetone-extractable fraction of the ink also brought about a similar cure rate for Meth A tumor. The delipidated ink enhanced the phagocytic activity of macrophages but no direct cytotoxicity was observed for the Meth A tumor cells. Hence it may be said that the anti-tumor activity of the delipidated ink was mainly due to the augmented cellular immunity in vivo. PMID- 9328865 TI - Glycation and inactivation of aspartate aminotransferase in diabetic rat tissues. AB - Glycated cytosolic aspartate aminotransferase was detected in the liver and kidney of streptozotocin diabetic rats using a boronate affinity column for adjacent cis-hydroxyl groups and an immunoblotting technique. The enzymatic activity and amount of immunoreactive substance were determined in the liver, kidney, and erythrocytes of diabetic and control rats. The ratio of enzymatic activity to the amount of enzyme was lower in diabetic rat tissue than in that in the control rats. It has been suggested that there is an inactive aspartate aminotransferase molecule in the tissues of diabetic rats. We therefore suggest that the cytosolic aspartate aminotransferase was inactivated in the diabetic rat tissues by a glycation reaction, accompanied by an impairment in glucose utilization. PMID- 9328866 TI - The inhibitory effect of vitamin E on arachidonic acid metabolism during the process of urethane-induced lung tumorigenesis in mice. AB - It is known that change in the arachidonic acid metabolism plays an important role in the development of tumors. This study was undertaken to understand the relationship of changes in lipoxygenase, cyclooxygenase and ornithine decarboxylase (ODC) to the inhibitory effect of vitamin E on urethane-induced lung tumorigenesis in mice. We analyzed the inhibitory effect of vitamin E on ornithine decarboxylase, cyclooxygenase and lipoxygenase activities at a promotion phase of lung tumorigenesis in mice. An increase in the ODC of urethane treated-mice and no significant change in the ODC of VE-treated mice were observed. An increase in the production of PGE2 and all HETES tested in the lungs of the urethane-treated mice was observed at week 8 after injection (promotion phase), showing a significant difference compared to the control group. Excessive vitamin E feeding during the initiation or promotion phases inhibited the increase in PGE2 and HETES produced by urethane treatment. These results suggest that the suppression of prostagrandin metabolism and ODC may be associated with the inhibitory effect of vitamin E against urethane-induced lung tumorigenesis. PMID- 9328867 TI - Conversion ratio of tryptophan to niacin in rats fed a vitamin B1-free diet. AB - The effect of a vitamin B1-free diet on the conversion ratio of tryptophan to niacin in rats was investigated using the current methods for determination of the intermediate metabolites. Rats were fed with diets with or without B1 for 33 days. The body weight gain and food intake in the B1-free group were almost the same as the control group for 10 days, but, they steeply dropped after that time and the conversion ratio of tryptophan to niacin began to increase. The ratio reached 7-fold that of the control group on the last day of the experiment. This finding seriously differed from the previous reports, which described that B1 was needed in the first part of tryptophan conversion to niacin and that the conversion ratio of tryptophan to niacin decreased in B1-deficient rats. Furthermore, the activity of tryptophan oxygenase, in which it was reported that B1 is required for the tryptophan catalytic reaction, did not decrease but increased even when the B1-free diet was fed. These results suggest that there is a very small possibility of the direct involvement of B1 in the conversion of tryptophan to niacin. PMID- 9328869 TI - Cognition and communication: referential strategies used by preschoolers with specific language impairment. AB - Ten children with specific language impairment and 10 children with normal language development were asked to describe objects so that a listener could select them. Each trial targeted two out of a group of three toys. The targeted objects were identical or were similar in size or color. Children in the two groups did not differ in referential success, although children in both groups found the size items more difficult. Content analysis of the messages did reveal differences in the referential strategies used most frequently. Children with specific language impairment were more likely to mention the attributes of each object separately, rather than to describe the characteristics common to a pair of objects. Children in both groups talked about separate objects more often when talking about size than about color or object type. Use of this strategy could indicate the effects of attentional capacity on children's solutions to communication tasks. PMID- 9328868 TI - Characterization of bovine heart sulfotransferase catalyzing the sulfation of tyrosine-containing peptides. AB - Using [35S]PAPS as the sulfate donor, we have detected a sulfotransferase from bovine heart which catalyzes the sulfation of tyrosine-containing peptides. The enzyme displayed optimal activity at pH 5.75 and 35 degrees C in a one-hour reaction. The addition of 10 mM Mn2+ or Co2+ to the reaction mixture increased the sulfotransferase activity by 3.4- and 3.5-fold, respectively. In contrast, the maximum increment stimulated by Mg2+ was only 1.75-fold at 15 mM concentration, and instead of exerting an enhancement effect, Ca2+ was found to be a potent inhibitor. The addition of 50 mM NaF to the reaction mixture resulted in an increase in sulfotransferase activity of 3.3-fold. The K(m) for 3' phosphoadenosine 5'-phosphosulfate (PAPS) was determined to be 2 microM at a constant 0.5 mM Boc-Glu-Asp-Tyr-Val. Among the 10 peptides tested as substrates, Boc-Glu-Asp-Tyr-Val and Boc-Asp-Asp-Tyr-Val provided the highest activities. PMID- 9328870 TI - The influence of sentence elicitation variables on children's speech production. AB - This study investigated the potential influence of adult-modeled sentences on the speech production of 15 children with speech delays of unknown origin. Two comparison tokens of target words containing sounds with inconsistently realized phonemes were sampled in picture descriptions elicited with and without adult modeled descriptive sentences. Ten listeners made forced-choice paired comparisons to identify the children's relatively more advanced word productions. From 205 total comparisons, listeners identified 130 word pairs that included one token more advanced than the other. Significantly more of the children's advanced word productions occurred in sentences elicited with an adult model sentence. Discussion considers theoretical and clinical perspectives of an assumption that variables facilitating children's language production may benefit speech production. PMID- 9328871 TI - Look who's talking: a prospective study of familial transmission of language impairments. AB - Language impairments have been hypothesized to have a genetic component. Previous studies of the familial aggregation of language impairments have relied on a retrospective approach based on parental/self-reported history of language development. This study examined familial aggregation prospectively, by investigating language acquisition and cognitive development in the younger siblings and offspring of individuals with well-defined language impairments. It was predicted that children with a positive family history for language impairments would be more likely to show delays in language acquisition than would age- and gender-matched controls. Similar delays were not expected in nonlinguistic domains, such as conceptual, gestural, or general cognitive development. Ten children with a positive family history and 10 age- and gender matched controls were tested. Analyses of linguistic and cognitive assessments at 16 to 26 months confirmed the predictions. Children with a family history of language impairments had lower receptive and expressive language scores than controls, with 50% of them scoring at least 1.5 SD below the mean for their age. At the same time, performance on a number of tasks that did not rely on language abilities did not differ as a function of family history. These results indicate that children with a positive family history for language impairments are at risk for language delay; the results also support a familial component to language impairments. PMID- 9328872 TI - A validity study of an implicit phonological awareness paradigm. AB - The purpose of this investigation was to examine the validity of a nonsense-word pairs paradigm as an implicit phonological awareness task. For this task one member of each nonsense-word-pair violated the rules of consonant combination in English (e.g., /integral kib/), and the other did not (e.g., /integral rib/). The subjects were required to choose the member of the pair that contained permissible consonant sequence(s). Eighty-one normally developing first- and second-graders were given the implicit phonological awareness task, 3 explicit phonological awareness tasks, 2 reading tasks, and a multisyllabic word production task. There were significant correlations between the implicit phonological awareness task and all of the experimental tasks, with the exception of one. Additionally, the implicit phonological awareness task was sensitive to developmental differences between the first- and second-grade readers. PMID- 9328873 TI - The ability of children with specific language impairment to access and participate in an ongoing interaction. AB - This study investigated the ability of 6 children with specific language impairment (SLI), ages 8;10 to 12;5 (years; months), to enter and participate in an ongoing dyadic interaction. Performance was compared to that of 6 chronological age-matched (CA) peers and 6 language-similar (LS) peers. All children in the LS and CA groups successfully accessed the interaction, and most did so quickly. Two children from the SLI group did not access, and the 4 remaining subjects required varying amounts of time to access. Following successful access, the triadic interactions of subjects were examined. The accessing children with SLI talked significantly less, were addressed significantly less, and collaborated less than either of the partners within their triads. Few significant differences were observed between LS or CA children and their partners. PMID- 9328874 TI - Naming difficulties in language-disabled children: preliminary findings with the application of the tip-of-the-tongue paradigm. AB - The "tip of the tongue" (TOT) paradigm in a picture-naming task was presented to 14 children with language disabilities (LD) and 14 children without language disabilities (ND). Although the two groups did not differ in the semantic information they had on words they could not fully retrieve, the LD children had less valid and more invalid phonological information. They also had fewer correct responses and spontaneous recalls, more "Don't Know" s (DK) and TOTs, and less accurate "feeling of knowing" (FOK) judgments. These results, demonstrating dissociation between the semantic and phonological levels of word representation, support a two-stage model of word retrieval. These findings are evidence in favor of a phonological treatment approach for naming problems in LD children. PMID- 9328875 TI - Reading and metaphonological outcomes in late talkers. AB - Children with a history of slow expressive language development (SELD) were followed to second grade, at which point outcomes in terms of speech, language, cognitive skills, reading achievement, and metaphonological performance were evaluated. Although there were some statistically significant differences between groups, children with a history of SELD generally performed within the normal range on the measures collected. Relations among speech, reading, and metaphonology in the SELD cohort appeared to operate in a manner similar to that seen in groups with typical language development. The implications of these findings for understanding the nature of specific language impairments and for treating early circumscribed language delays are discussed. PMID- 9328876 TI - Japanese speakers of American English: competence with connectives in written language. AB - Competence with literate connectives (e.g., conversely, similarly, moreover) is important for academic success, particularly in university settings. This study examined the ability of Japanese speakers of American English (mean age = 23 years) who were attending an American university to use and understand connectives in written language. Compared to a control group of age-matched native English-speakers attending the same university, the Japanese students had difficulty with the words. Suggestions are offered on how speech-language pathologists might facilitate competence with connectives in adults who are non native speakers of English. PMID- 9328877 TI - Long-term phonatory instability in individuals with multiple sclerosis. AB - This paper uses a new approach to describe and quantify the long-term phonatory instability of speakers with MS. Sustained vowel phonations of 20 individuals with a definite diagnosis of multiple sclerosis (MS) and 20 age- and gender matched individuals with normal speech were recorded. The phonations were f0 and intensity analyzed and subjected to spectral analysis using the Fast Fourier Transform. Three methods for analyzing the instabilities are presented, compared, and related to perceptual judgments: (a) coefficients of variation, (b) magnitude based analysis of spectral energy, and (c) frequency-based analysis of spectral components. All measures reliably distinguished between individuals with MS and persons with normal speech. A single factor based on a linear discriminant analysis of the frequency-based measures was especially useful in distinguishing these groups. Critical frequency bands of instability, corresponding to wow (1-2 Hz), tremor (around 8 Hz), and flutter (17-18 Hz), distinguished the MS group from those of the control group. PMID- 9328879 TI - Development of a two-stage procedure for the automatic recognition of dysfluencies in the speech of children who stutter: II. ANN recognition of repetitions and prolongations with supplied word segment markers. AB - This program of work is intended to develop automatic recognition procedures to locate and assess stuttered dysfluencies. This and the preceding article focus on developing and testing recognizers for repetitions and prolongations in stuttered speech. The automatic recognizers classify the speech in two stages: In the first the speech is segmented and in the second the segments are categorized. The units segmented are words. The current article describes results for an automatic recognizer intended to classify words as fluent or containing a repetition or prolongation in a text read by children who stutter that contained the three types of words alone. Word segmentations are supplied and the classifier is an artificial neural network (ANN). Classification performance was assessed on material that was not used for training. Correct performance occurred when the ANN placed a word into the same category as the human judge whose material was used to train the ANNs. The best ANN correctly classified 95% of fluent, and 78% of dysfluent words in the test material. PMID- 9328878 TI - Development of a two-stage procedure for the automatic recognition of dysfluencies in the speech of children who stutter: I. Psychometric procedures appropriate for selection of training material for lexical dysfluency classifiers. AB - This program of work is intended to develop automatic recognition procedures to locate and assess stuttered dysfluencies. This and the following article together, develop and test recognizers for repetitions and prolongations. The automatic recognizers classify the speech in two stages: In the first, the speech is segmented, and, in the second, the segments are categorized. The units that are segmented are words. Here assessments by human judges on the speech of 12 children who stutter are described using a corresponding procedure. The accuracy of word boundary placement across judges, categorization of the words as fluent, repetition or prolongation, and duration of the different fluency categories are reported. These measures allow reliable instances of repetitions and prolongations to be selected for training and assessing the recognizers in the subsequent paper. PMID- 9328880 TI - Acoustic measures of temporal intervals across speaking rates: variability of syllable- and phrase-level relative timing. AB - It has been suggested previously that at least some levels of the temporal organization for speech production are characterized by proportional timing. The proportional timing model maintains that the duration of temporal intervals within a sequence would remain proportionally invariant across changes in overall duration of the sequence. In order to test this hypothesis for the acoustic level of speech production, 18 women produced three trials of the utterance "Buy Bobby a poppy" at each of three speaking rates (i.e., slow, normal, fast). Acoustically derived temporal intervals were paired to form ratios reflecting either syllable level or phrase-level relative timing. Findings indicated that ratios of temporal intervals at both the syllable-level and phrase-level did not remain invariant across speaking rates. Rather, statistically significant changes in the relative duration of both types of intervals were observed as a function of overall rate of production. For most of the obtained ratios, the direction of these changes was highly consistent across individual subjects. PMID- 9328881 TI - On the registration of time and the patterning of speech movements. AB - In order to study speech coordination we frequently average kinematic and other physiological signals. The averages are assumed to be more representative of the underlying patterns of production than individual records. In this note we outline different approaches to averaging and present a new nonlinear normalization technique that offers better information than ensemble averaging, linear normalization, or feature alignment methods. We suggest that this technique provides a clear estimation of pattern shape while preserving information on the variation over time. PMID- 9328882 TI - A system for three-dimensional visualization of human jaw motion in speech. AB - With the development of precise three-dimensional motion measurement systems and powerful computers for three-dimensional graphical visualization, it is possible to record and fully reconstruct human jaw motion. In this paper, we describe a visualization system for displaying three-dimensional jaw movements in speech. The system is designed to take as input jaw motion data obtained from one or multi-dimensional recording systems. In the present application, kinematic records of jaw motion were recorded using an optoelectronic measurement system (Optotrak). The corresponding speech signal was recorded using an analog input channel. The three orientation angles and three positions that describe the motion of the jaw as a rigid skeletal structure were derived from the empirical measurements. These six kinematic variables, which in mechanical terms account fully for jaw motion kinematics, act as inputs that drive a real-time three dimensional animation of a skeletal jaw and upper skull. The visualization software enables the user to view jaw motion from any orientation and to change the viewpoint during the course of an utterance. Selected portions of an utterance may be replayed and the speed of the visual display may be varied. The user may also display, along with the audio track, individual kinematic degrees of freedom or several degrees of freedom in combination. The system is presently being used as an educational tool and for research into audio-visual speech recognition. Interested researchers may obtain the software and source code free of charge from the authors. PMID- 9328883 TI - Estimating phonation threshold pressure. AB - Phonation threshold pressure (PTP) is the minimum subglottal pressure required to initiate vocal fold oscillation. Although potentially useful clinically, PTP is difficult to estimate noninvasively because of limitations to vocal motor control near the threshold of soft phonation. Previous investigators observed, for example, that trained subjects were unable to produce flat, consistent oral pressure peaks during/pae/syllable strings when they attempted to phonate as softly as possible (Verdolini-Marston, Titze, & Druker, 1990). The present study aimed to determine if nasal airflow or vowel context affected phonation threshold pressure as estimated from oral pressure (Smitheran & Hixon, 1981) in 5 untrained female speakers with normal velopharyngeal and voice function. Nasal airflow during /p/occlusion was observed for 3 of 5 participants when they attempted to phonate near threshold pressure. When the nose was occluded, nasal airflow was reduced or eliminated during /p/;however, individuals then evidenced compensatory changes in glottal adduction and/or respiratory effort that may be expected to alter PTP estimates. Results demonstrate the importance of monitoring nasal flow (or the flow zero point in undivided masks) when obtaining PTP measurements noninvasively. Results also highlight the need to pursue improved methods for noninvasive estimation of PTP. PMID- 9328884 TI - Effect of altered auditory feedback on people who stutter during scripted telephone conversations. AB - The effect of altered auditory feedback (AAF) conditions on stuttering during scripted telephone conversations was investigated. Nine adult participants made 15 scripted telephone calls to business in New York City. Alterations in the participants' auditory feedback signal were generated by a commercially available digital signal processor (Casa Futura Technologies Desktop Fluency System Model BTD-400) that shifted participants' speech one-half octave down in frequency, produced a 50-ms delay, or produced non-altered auditory feedback. The AAF effects produced by the digital signal processor were not perceived by the recipients of the telephone calls. The proportion of stuttering events per scripted telephone conversations were significantly reduced in the AAF conditions relative to the non-altered auditory feedback condition (p = .0004). Stuttering frequency was reduced by 55% and 60% for the FAF and DAF, respectively. These findings demonstrate the applicability of this technology to situations of daily living involving telephone use. PMID- 9328885 TI - Multisensory speech perception of young children with profound hearing loss. AB - The contribution of a two-channel vibrotactile aid (Trill VTA 2/3, AVR Communications LTD) to the audiovisual perception of speech was evaluated in four young children with profound hearing loss using words and speech pattern contrasts. An intensive, hierarchical, and systematic training program was provided. The results show that the addition of the tactile (T) modality to the auditory and visual (A+V) modalities enhanced speech perception performance significantly on all tests. Specifically, at the end of the training sessions, the tactile supplementation increased word recognition scores in a 44-word, closed-set task by 12 percentage points; detection of consonant in final position by 50 percentage points; detection of sibilant in final position by 30 percentage points; and detection of voicing in final position by 25 percentage points. Significant learning over time was evident for all test materials, in all modalities. As expected, fastest learning (i.e., smallest time constants) was found for the AVT condition. The results of this study provide further evidence that sensory information provided by the tactile modality can enhance speech perception in young children. PMID- 9328886 TI - The effect of communication mode on the development of phonemic awareness in prelingually deaf students. AB - Two groups of prelingually deaf children and a hearing control group participated in an experiment examining the effect of communication mode on the development of phonemic awareness. Sixteen of the deaf students (mean grade 6.9) were trained orally, using spoken language as their principal means for communication at home and at school. Another 16 deaf students (mean grade 6.9), all of them deaf children of deaf parents, acquired sign language as their primary language. The mean grade of the hearing control group was 6.5. The performance of the two deaf groups indicates that permanent auditory deprivation leads to substantially reduced phonemic awareness but does not entirely block its development. Contrary to expectation, the development of phonemic awareness in individuals with impaired hearing was not significantly affected by their preferred communication mode. Results further suggested that, for deaf individuals with excellent skills in sign language, the functional impairment caused by prelingual deafness may be restricted to the processing of phonological information. PMID- 9328887 TI - COAE thresholds: 1. Effects of equal-amplitude versus subtraction methods. AB - Although research has demonstrated that click-evoked otoacoustic emissions (COAEs) elicited by high-level stimuli are useful for identifying hearing loss, the ability of COAEs to predict behavioral thresholds has not been adequately tested. Results of studies comparing COAE thresholds and behavioral thresholds have been equivocal, perhaps due to the need for a more rigorous approach to COAE threshold estimation. The present study was designed to address several methodological concerns in COAE threshold testing, particularly the effects of two methods of stimulus presentation on COAE testing and threshold calculation. In an attempt to make COAE threshold estimation consistent across participants, COAE threshold calculations were based on mean noise floor levels across participants. COAE and noise floor levels were measured in 15 participants using both equal-amplitude clicks and a subtraction method. Broadband COAEs were analyzed into 1/3 octave bands, so that input/output functions could be examined and COAE thresholds could be calculated for each 1/3 octave band. Comparison of the two stimulus methods indicated several differences. Mean noise floor levels for the equal-amplitude method were approximately 6 dB lower than those measured for the subtraction method across frequency. In many cases COAEs evoked using the equal-amplitude method were higher in amplitude than those evoked using the subtraction method. COAE thresholds measured using the equal-amplitude click stimuli were significantly lower than those measured using the subtraction method. The significantly higher thresholds obtained using the subtraction method may be attributed in part to the reduction of COAE amplitude by the subtraction procedure, and not merely to the higher noise level. Slopes of the input/output functions were not significantly different between the two stimulus methods. These results suggest that the equal-amplitude method is preferable for COAE threshold testing because lower noise floor and larger amplitude COAEs may be obtained in the same test time. PMID- 9328888 TI - The perception of internal circuit noise in hearing aids by listeners with normal hearing. AB - Internal circuit noise in hearing aids is distracting to a listener and, if loud enough, may interfere with intelligibility, either by direct masking of weak components of speech or through the generation of undesired intermodulation products, which can also act as a source of masking. The objective characteristics of noise may be measured; however, wearers of hearing aids often differ in their subjective reporting of the perceived characteristics of the internal noise. This study reports on the results for four listeners with normal hearing of matching pitch and amplitude to the internal noise generated within a series of hearing aids. Results of these experiments showed that the listeners (a) primarily matched the perceived pitch of the noise to the frequency of their most sensitive hearing, and (b) matched the perceived level of the noise approximately to the total SPL noise level. PMID- 9328889 TI - Effects of age, speech rate, and type of test on temporal auditory processing. AB - Cognitive slowing that accompanies aging may be reflected in temporal aspects of auditory processing. The purpose of this study was to investigate the effects of age, type of test, and rate of speech on temporal auditory processing. Listeners were divided into three groups: young (25- to 35-year-olds), middle aged (45- to 55-year-olds), and older (65- to 75-year-olds). A method of time compression known as Synchronized Overlap Add (SOLA) was used to increase the rate of speech. This method provides a high-quality speech signal and limits the distortions that may confound the temporal effects on time-compressed tests of speech intelligibility. Listeners performed four speech understanding tasks: sentence repetition, sentence intelligibility rating, connected discourse intelligibility rating, and connected discourse comprehension question and answers at three time compression rates (60%, 70%, and 80%). Although the older group performed more poorly on all tests, only the connected discourse intelligibility rating test was sensitive to age differences among all three groups. This difference did not appear to increase with rate increases but was present only at the 70% compression rate. In addition, variability was especially high in the oldest group of participants. PMID- 9328891 TI - Hearing sensitivity in newborns estimated from ABRs to bone-conducted sounds. AB - This study focused on the problem of estimating hearing sensitivity in newborns from auditory brainstem responses (ABRs) evoked by clicks and 500 Hz and 4000 Hz tonebursts presented by a bone-conduction (BC) oscillator. The effects of acoustic energy transmitted to the ear canal, gender, and ear differences were also investigated. ABR thresholds for BC stimuli were 56, 52, and 53 dB (re 1 microN) or -5, -14, and 0 dB nHL (re adult psychophysical threshold) for click and 500 Hz and 4000 Hz tonebursts, respectively. For newborns, ear canal SPLs generated by the BC stimuli were as much as 21 dB greater than those in adults. Gender-related threshold differences were significant, with female infants having lower thresholds than males; however, ear differences were not. The findings of this study can be used to set appropriate BC stimulus levels for screening or assessment of newborns. PMID- 9328890 TI - Speech recognition as a function of the number of electrodes used in the SPEAK cochlear implant speech processor. AB - Speech recognition was measured in listeners with the Nucleus-22 SPEAK speech processing strategy as a function of the number of electrodes. Speech stimuli were analyzed into 20 frequency bands and processed according to the usual SPEAK processing strategy. In the normal clinical processor each electrode is assigned to represent the output of one filter. To create reduced-electrode processors the output of several adjacent filters were directed to a single electrode, resulting in processors with 1, 2, 4, 7, 10, and 20 electrodes. The overall spectral bandwidth was preserved, but the number of active electrodes was progressively reduced. After a 2-day period of adjustment to each processor, speech recognition performance was measured on medial consonants, vowels, monosyllabic words, and sentences. Performance with a single electrode processor was poor in all listeners, and average performance increased dramatically on all test materials as the number of electrodes was increased from 1 to 4. No differences in average performance were observed on any test in the 7-, 10-, and 20-electrode conditions. On sentence and consonant tests there was no difference between average performance with the 4-electrode and 20-electrode processors. This pattern of results suggests that cochlear implant listeners are not able to make full use of the spectral information on all 20 electrodes. Further research is necessary to understand the reasons for this limitation and to understand how to increase the amount of spectral information in speech received by implanted listeners. PMID- 9328892 TI - Interaural time effects on the frequency-following response. AB - Frequency-following responses (FFRs) were recorded to evaluate differences between monaural and binaural waveforms and waveforms evoked by stimuli with interaural time disparities. Eight normal-hearing adult females served as subjects. The stimuli were monaural and binaural 450-Hz tonebursts at 65 and 60 dB SL and interaural time differences of 0 and 660 microseconds, respectively. Normalized amplitudes and periodicities of FFR waveforms within and between subjects were compared. The results showed asymmetric FFR to the various stimuli used in this study. Binaural FFR waveforms were greater than monaural but smaller than summed monaural FFRs. Binaural FFR amplitudes evoked by a zero time difference were greater than amplitudes evoked by a 660-microseconds difference. Additionally, tight phase-locked periodicities were evoked in the FFR monaurally and binaurally. The averaged FFR periodicity to all stimulus conditions from all subjects was 2.29 msec, differing only 6.8 microseconds from the period of the 450-Hz stimulus. In contrast, monaural and binaural neurons in the lower brain stem typically exhibit much less synchroneities to low-frequency tones than the FFR. These data provide evidence that the FFR is not simply a sum of neuronal action potentials. The findings suggest instead the presence of brainstem neuronal networks. Such putative neuronal ensembles apparently maintain a closer correspondence to the period of a low-frequency sound, whether monaural or binaural, than the discharge patterns of single neurons. PMID- 9328893 TI - Aural rehabilitation and graduate audiology programs. AB - The quantity and quality of aural rehabilitation training that audiology graduate students receive versus the current services available to those students regarding work with the hearing impaired, cochlear implant recipients, patients with vestibular disorders, patients with central auditory processing disorders, and patients suffering from tinnitus were examined. Forty ASHA-accredited colleges and universities were contacted by telephone and audiology faculty members were surveyed. The results of the survey may provide valuable information for identifying and addressing shortfalls in aural rehabilitation programs. PMID- 9328894 TI - Consonant perception in quiet: effect of increasing the consonant-vowel ratio with compression amplification. AB - Single-channel syllabic compression amplification may increase the consonant vowel ratio (CVR). This study was conducted to investigate the effect of CVR increases, associated with syllabic compression, on consonant perception in quiet for people with normal hearing and those with sensorineural hearing impairment. Fifteen hearing-impaired and 15 normal-hearing older people were assessed with different versions of the Nonsense Syllable Test, which had been recorded with linear and compression amplification (compression ratio = infinity). Overall scores did not differ significantly with type of amplification for both subject groups. Analysis of classes of sounds revealed the differential effect of type of amplification for the subject groups. This study highlights the need for audiologists to be aware that applying amplification that raises the level of consonants in relation to vowels is not always beneficial for people with hearing impairment, as the evidence indicates that CVR may be a cue to the perception of some consonants. PMID- 9328895 TI - Application of a stimulus spectral calibration routine to click evoked otoacoustic emissions. AB - This study examined the influence of a calibrated transient stimulus on click evoked otoacoustic emissions (CEOAEs). The calibration procedure produced a spectrally uniform stimulus (1-8 kHz) at the plane of the ear probe that was very similar among individuals. However, the calibrated signal reduced the overall level and repeatability of the CEOAE, probably due to the minimization of the energy peak at 2 kHz, which is enhanced in the uncalibrated signal. The amplitude of CEOAEs obtained with the calibrated signal was less variable among individuals compared to CEOAEs obtained with the uncalibrated signal. PMID- 9328896 TI - Comparison of the intersubject and intrasubject variability of exogenous and endogenous auditory evoked potentials. AB - The variability, both between subjects (intersubject variability) and within subjects (intrasubject variability), of the P300 event-related potential was compared to that of early and middle latency components using coefficients of variation. For each of five waveform components, 12 amplitude measures (two trials on each of six measurement occasions) and 12 latency measures were obtained for each of eight subjects. P300 intersubject variability was comparable to that of the most stable early components (ABR waves III and V), both in terms of amplitude and latency. Intrasubject variability was considerably greater for P300 than for the early components; even so, multiple waveforms collected from the same subject tended to fit within an envelope bounded by +/- 1 standard deviation form the mean (averaged) response to oddball stimuli. The variability of MLR component Pa exceeded that of all other components. PMID- 9328897 TI - Comparison of performance with frequency transposition hearing aids and conventional hearing aids. AB - Four experienced hearing aid users were evaluated using a frequency transposition (TranSonic) hearing system. Following a trial period, the Abbreviated Profile of Hearing Aid Performance (APHAB) and a variety of speech audiometric measures were used to compare the frequency transposition fitting with each subject's conventional hearing aids. A single-subject study design with a series of repeated measures permitted statistical analysis of differences in performance with the various amplification strategies. Two of the four subjects demonstrated statistically significant benefit with the frequency transposition device. Results show the efficacy of frequency transposition in improving speech understanding and quality of life in some individuals with severe-to-profound hearing loss. Overall, results suggest the need for evaluating the benefit of frequency transposition on an individual basis. PMID- 9328926 TI - Mannitol therapy revisited (1940-1997). PMID- 9328925 TI - Pregnancy in renal disease. PMID- 9328927 TI - Mutations in the CLCN5 gene in Japanese patients with familial idiopathic low molecular-weight proteinuria. AB - Familial idiopathic low-molecular-weight proteinuria (FILMWP) is a renal proximal tubulopathy that occurs predominantly in males. FILMWP is characterized by mild proteinuria consisting of low-molecular-weight proteinuria, aminoaciduria and relatively conserved renal function, but without rickets. To determine whether FILMWP is related to the CLCN5 gene, which is responsible for Dent's disease and two related disorders, we analyzed the CLCN5 gene from four Japanese families with FILMWP. We identified two novel mutations: one was a single base insertion at codon 520 serine in exon 10 and the other was a single base deletion at codon 403 tyrosine in exon 8. These mutations caused a shift in the reading frame, resulting in synthesis of truncated CLC5 proteins that lacked 220 (29%) and 314 (42%) amino acids, respectively. These mutations were demonstrated to cosegregate with the disease in two families, respectively. We conclude that the CLCN5 gene is responsible for this proximal renal tubulopathy in some Japanese families and that FILMWP is possibly a variant of Dent's disease. PMID- 9328928 TI - Protein kinase C in the developing kidney: isoform expression and effects of ceramide and PKC inhibitors. AB - Protein kinase C (PKC) is a serine/threonine kinase recognized as a key enzyme in signal transduction mechanisms in various biological processes. During development, PKC is involved in the regulation of growth and differentiation. In mature tissue PKC is important for homeostatic functions. We studied PKC with regard to expression and effects on differentiation, growth and apoptosis in the developing kidney. Using in situ hybridization, we demonstrate age-dependent expression of PKC alpha, PKC delta, PKC zeta and PKC lambda during fetal and postnatal kidney development. The endogenous sphingolipid product ceramide, as well as specific PKC inhibitors, disturbed nephron formation and induced apoptosis in organ cultures of E13 kidneys. In primary cell cultures of proximal tubule cells, ceramide and the specific PKC inhibitors induced apoptosis. In conclusion, PKC alpha, PKC delta, PKC zeta and PKC lambda are expressed in an age dependent pattern during kidney development. Inhibition of PKC disturbs nephron formation, inhibits growth and induces apoptosis in the developing kidney. The findings suggest that PKC plays an important role in regulating normal kidney growth and differentiation. PMID- 9328929 TI - Mutations of CLCN5 in Japanese children with idiopathic low molecular weight proteinuria, hypercalciuria and nephrocalcinosis. AB - The annual urinary screening of Japanese children above three years of age has identified a progressive renal tubular disorder characterized by low molecular weight proteinuria, hypercalciuria and nephrocalcinosis. The disorder has been observed in over 60 patients and has a familial predisposition. Mutations of a renal chloride channel gene, CLCN5, have been reported in four such families, and we have undertaken studies in additional patients from 10 unrelated, non consanguineous Japanese families to further characterize such CLCN5 mutations and to ascertain their prevalence. CLCN5 abnormalities we identified in 7 of the 10 unrelated patients and consisted of 5 mutations (2 nonsense, 1 frameshift and 2 missense), 1 deletion and 1 silent polymorphism. A clustering of these mutations in CLCN5 exons 8 and 10 was observed. Over 80% of the CLCN5 mutations could be readily detected by single stranded conformational polymorphism (SSCP) analysis, thereby providing a useful mutation screening method. Our results, which indicate that over 70% of Japanese patients with this renal tubulopathy have CLCN5 mutations, will help in the genetic and clinical evaluation of children at risk from this disorder. PMID- 9328930 TI - Cytoprotective effects of adrenomedullin in glomerular cell injury: central role of cAMP signaling pathway. AB - Activation of cAMP signaling pathway was shown to inhibit some pathobiologic processes in mesangial cells (MC). We investigated whether adrenomedullin (ADM), a potent agonist of adenylate cyclase, is synthesized in MC and whether it can, via cAMP, suppress the generation of reactive oxygen metabolites (ROM) and proliferation of cells in glomeruli. With the use of an immunohistologic technique ADM was detected in mesangial and microvascular areas of rat glomeruli. MC grown in primary culture synthesized ADM, and the synthesis was stimulated by TNF alpha and IL-1 beta but not by PDGF and EGF. ADM inhibited ROM generation in MC dose-dependently and caused in situ activation of protein kinase A (PKA). In macrophages (cell line J774) ROM generation was about four times higher than in MC and was inhibited by ADM in a similar way as in MC. The rate of MC proliferation, measured by [3H]-incorporation, and the activity of mitogen activated protein kinase (MAPK) stimulated by PDGF and EGF were dose-dependently inhibited by ADM; the maximum inhibition (at 10 nM ADM) was about -80%. Mitogenesis of MC and MAPK activity when stimulated to a similar extent by endothelin (ET-1) was inhibited by ADM to a significantly (P < 0.01) lesser degree (-30%). Further, ADM inhibited PDF-stimulated mitogenesis and activation of MAPK in cultured vascular smooth muscle cells (VSMC). The inhibition of PDGF activated MAPK by ADM in VSMC was reversed by the protein kinase A (PKA) inhibitor, H89. Taken together, results indicate the adrenomedullin (ADM) generated in mesangial cells (MC) can suppress, via activation of the cAMP protein kinase A (PKA) signaling pathway, reactive oxygen metabolites (ROM) generation in MC and infiltrating macrophages as well as mitogen-activated protein kinase (MAPK)-mediated mitogenesis in MC and vascular smooth muscle cells (VSMC). We suggest that introglomerular ADM may serve as a cytoprotective autoacoid that suppresses pathobiologic processes evoked by immuno-inflammatory injury of glomeruli. PMID- 9328931 TI - Rapid communication. Enalapril decreases nuclear factor kappa B activation in the kidney with ureteral obstruction. AB - The transcription factor nuclear factor kappa B (NF-kappa B) controls a number of genes associated with tissue inflammation and has been shown to be activated in the kidney with ureteral obstruction. In this investigation, we further explored NF-kappa B activation in the kidney cortex of rats with unilateral ureteral obstruction. Electrophoretic mobility shift assays combined with antibody supershift/depletion demonstrated that NF-kappa B subunits p50, p52, c-rel, p65 (RelA) and RelB were all activated. Immunocytochemical analysis using an antibody to the p50 subunit demonstrated activation occurring predominantly in nuclei of tubular cells. Treatment of animals with unilateral ureteral obstruction with an oral ACE inhibitor significantly decreased NF-kappa B activation. This suggests that the antiinflammatory effect of ACE inhibitors in renal disease is in part due to a blunting of NF-kappa B activation. PMID- 9328932 TI - Systemic autoimmune nephritogenic components induce CSF-1 and TNF-alpha in MRL kidneys. AB - MRL-Faslpr mice are an appealing strain to understand the importance of cytokines in the pathogenesis of autoimmune renal destruction, since injury is rapid and predictable. Colony stimulating factor 1 (CSF-1) and tumor necrosis factor alpha (TNF-alpha) are detected in the kidney and circulation prior to renal injury and continue to increase with progressive renal damage in MRL-Faslpr, Fas deficient mice, but not the congenic Fas intact MRL-(++) strain. Delivery of CSF-1, but not TNF-alpha, into the kidney via gene transfer incites local renal injury. By comparison, dual gene transfer of CSF-1 and TNF-alpha incites autoimmune renal injury that is far more severe than CSF-1 alone. The purpose of this study was to establish whether CSF-1 and TNF-alpha incites autoimmune renal injury that is far more severe than CSF-1 alone. The purpose of this study was to establish whether CSF-1 and TNF-alpha expression in the kidney of MRL-Faslpr mice induced by a circulating stimulant resulted from a primary defect in the kidney. Therefore, we transplanted (Tx) a MRL-(++) kidney without CSF-1, TNF-alpha and renal injury into an MRL-Faslpr recipient after removing nephritic kidney expressing CSF-1 and TNF-alpha. The Tx kidneys were examined after 2, 4, 5, 6, and 12 weeks. CSF-1 and TNF-alpha were rapidly induced in the MRL-(++) Tx kidney. CSF-1 and TNF-alpha were evident by two weeks and continually increased for 12 weeks post transplantation. Within several weeks, the rapid expansion of M phi and T cells and induction of glomerulonephritis and interstitial nephritis in the MRL-(++) Tx kidney was similar to the age-matched native MRL-Faslpr kidney. In conclusion, we have constructed an experimental transplantation system that can explore whether cytokine expression in the kidney induced by a circulating stimulant is a result of a primary defect in the kidney. Using this approach, we established that the MRL-Faslpr kidney is not defective, but rather a circulating stimulant in the MRL Faslpr mouse can induce CSF-1, TNF-alpha and renal injury in a normal MRL-(++) kidney. Thus, we exclude an intrinsic defect in the MRL-Faslpr kidney as the pathogenic mechanism responsible for tissue damage. We suggest purging the circulation of molecules that induce CSF-1 and TNF-alpha as a therapeutic strategy for autoimmune renal injury. PMID- 9328933 TI - Decreased expression of mitochondrial-derived H2O2 and hydroxyl radical in cytoresistant proximal tubules. AB - Increased production of reactive oxygen metabolites (ROM) can contribute to the initiation phase of nephrotoxic and ischemic acute renal failure (ARF). However, whether altered ROM expression also exists during the maintenance phase of ARF has not been adequately assessed. Since diverse forms of tubular injury can initiate a "cytoresistant state," this study tested whether a down-regulation of ROM expression might develop in the aftermath of acute tubular damage, potentially limiting renal susceptibility to further attack. To test this hypothesis, rats were subjected to either mild myohemoglobinuria (glycerol injection) or bilateral ureteral obstruction and 24 hours later, cytoresistant proximal tubular segments (PTS) were isolated to assess ROM expression. PTS from sham operated rats were used to establish normal values. Both sets of cytoresistant PTS manifested approximately 75% reductions in H2O2 levels, as assessed by the phenol red/horseradish peroxidase technique (P < 0.01 to 0.001). A 40% reduction in hydroxyl radical (.OH) levels was also observed (salicylate trap method), thereby substantiating decreased oxidant stress in cytoresistant PTS. Catalase, glutathione peroxidase, and free iron levels were comparable in control and cytoresistant PTS, suggesting that decreased H2O2 production (such as by mitochondria) was the cause of the decreased oxidant stress. To test this latter hypothesis, H2O2 expression by control and cytoresistant PTS was assessed in the presence of respiratory chain inhibitors. Although site 1 and site 3 inhibition markedly suppressed H2O2 production in control PTS, they had no impact on H2O2 production in cytoresistant PTS, implying that production at these sites was already maximally suppressed. Correlates of the decreased mitochondrial H2O2 production were improvements in cell energetics (increased ATP/ADP ratios with Na ionophore treatment) and approximately 40 to 90% increases in PTS/renal cortical glutathione content. We conclude that: (1) proximal tubule H2O2/.OH expression can be downregulated during the maintenance phase of ARF; (2) this seemingly reflects a decrease in mitochondrial ROM generation; and (3) the associated improvements in glutathione content and/or cellular energetics could conceivably contribute to a post-injury cytoresistant state. PMID- 9328934 TI - Overexpression of ecto-5'-nucleotidase promotes P-glycoprotein expression in renal epithelial cells. AB - P-glycoprotein (P-gp), responsible for multidrug resistance (MDR) of tumoral cells, is also expressed in apical membranes of normal epithelial cells, among which are proximal tubular cells. Ecto-5'-nucleotidase (5'Nu), co-located with P gp in renal brush border membranes, could be instrumental in the expression of MDR phenotype. P-gp activity [assessed by rhodamine 123 (R123) and [3H]vinblastine (3H-VBL) accumulation] was evaluated in MDCK cell lines in which human 5'Nu was expressed at different levels after retroviral infection: MDCK 5'NU/- cells with a low 5'Nu activity (Vmax < 2 pmol/mg protein/min) and MDCK 5'NU/+ cells, which expressed a high level of 5'Nu (Vmax 150 +/- 18.5 pmol/mg protein/min). MDCK-5'NU/- cells did not display functional expression of MDR. In MDCK-5'NU/+ cells, R123 and 3H-VBL accumulation was significantly lower than in MDCK-5'NU/- cells and was dramatically enhanced by P-gp inhibitors. This high P gp activity in MDCK-5'NU/+ cells was confirmed by their resistance to colchicine (measured by LDH release and MTT assay) as compared to MDCK-5'NU/- and was accounted for by increased membrane expression of P-gp assessed by Western blot. Neither AMP nor adenosine, the substrate and the product of 5'Nu, respectively, affected P-gp activity. Inhibition of 5'Nu with alpha beta-methylene-adenosine diphosphate (alpha beta MADP) or with a blocking anti-5'Nu antibody (1E9) did not blunt MDR expression in MDCK-5'NU/+ cells. Conversely, the anti-5'Nu antibody 5F/F9, which did not block the enzymatic site, induced a decrease of P-gp activity. Further, incubation of MDCK-5'NU/- cells with conditioned medium from MDCK-5'NU/+ cells, which contained significant amounts of released 5'Nu, induced MDR phenotype. IN CONCLUSION: (i) expression of ecto-5'Nu promotes multidrug resistance (MDR) activity in renal epithelial cells by enhancement of P-gp expression; (ii) this effect does not involve enzymatic activity of 5'Nu; (iii) supernatants of cells that express 5'Nu conferred P-gp activity to 5'Nu negative cells. PMID- 9328935 TI - HMG-CoA reductase inhibitors induce apoptosis in mouse proximal tubular cells in primary culture. AB - Renal cyst formation in polycystic diseases or after nephron reduction is attributed to enhanced tubular cell proliferation with unbalanced cell death. The induction of tubular cell death could be effective to reduce renal cyst formation. In this study, we examined the effects of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors on apoptosis in mouse proximal tubular (MPT) cells in primary culture. After treatment with HMG-CoA reductase inhibitors, the extracted DNA was analyzed by gel-electrophoresis under ultraviolet light. Apoptosis was evaluated quantitatively by estimating the ratio of fragmented DNA over intact DNA. For morphologic studies, cells were stained with Hoechst 33,258. DNA ladder pattern of 200 kDa typical of apoptosis and significant increase in DNA fragmentation were seen after 24 hours of treatment with lovastatin, a HMG-CoA reductase inhibitor. Staining with the Hoechst dye revealed cleavage of nucleus into pieces under the same condition. Geranylgeranylpyrophosphate (20 microM) and mevalonate (500 microM) completely reversed the effect of lovastatin, while farnesylpyrophosphate (20 microM) partially reversed it. Other products of HMG-CoA pathway such as cholesterol, ubiquinone, dolichol, and isopentenyladenine had no effect. Perillic acid and alpha-hydoxyfarnesylphosphonic acid, isoprenylation inhibitors, induced apoptosis of the cells. A treatment with lovastatin caused actin filament disruption. Cytochalasin D, an inhibitor of actin polymerization, induced apoptosis. Interleukin-1 beta-converting enzyme inhibitor II, a protease inhibitor, had no effect on the apoptosis induced by either HRI or cytochalasin D. The present study suggests that in mouse proximal tubules, HMG-CoA reductase inhibitors induce apoptosis via inhibition of isoprenoid production, and disruption of actin filaments may play a role in the apoptosis induction. PMID- 9328936 TI - Exposure of human renal proximal tubular cells to glucose leads to accumulation of type IV collagen and fibronectin by decreased degradation. AB - Thickening and reduplication of the tubular basement membrane has been reported as an early event in diabetic nephropathy. In the current study we examined the effects of elevated D-glucose concentrations on human proximal tubular (HPTC) type IV collagen and fibronectin turnover. Incubation of confluent growth arrested HPTC with 25 mM D-glucose led to accumulation of both type IV collagen and fibronectin. This effect was maximal at 48 hours and represented a sevenfold increase for fibronectin (N = 4, P = 0.04), and a threefold increase for type IV collagen (N = 3, P = 0.03) over cells exposed to 5 mM D-glucose controls. This increase was not dependent on new gene transcription for either protein. Tissue inhibitor of metalloproteinases (TIMP 1 + TIMP 2) were induced following addition of 25 mM D-glucose, but not when cells were exposed to 5 mM D-glucose. Twenty four hours after the addition of 25 mM D-glucose there was an eightfold increase in TIMP 1 (P = 0.009, N = 4), and a tenfold increase in TIMP 2 levels (P = 0.003, N = 4), over the control values for both inhibitors. The increase in both TIMP 1 and TIMP 2 in response to 25 mM D-glucose was abrogated in a dose dependent manner by the aldose reductase inhibitor sorbinil. Gelatin-substrate gel zymography showed increased activity of gelatinase A, but not of gelatinase B in response to the addition of 25 mM D-glucose to HPTC. The induction of gelatinase A was accompanied by increased gelatinase A mRNA expression, which was inhibited both by protein kinase C (PKC) depletion using PMA pre-treatment, and by the addition of a PKC inhibitor. These data demonstrate that the glucose-induced accumulation of type IV collagen and fibronectin is unrelated to increased gene transcription, but may involve alterations in the degradative pathway of these basement membrane constituents. Furthermore, the data demonstrate that glucose may simultaneously activate two intracellular pathways (the polyol pathway and a PKC dependent activation pathway), which are involved in mediating separate, complementary effects on cell function. PMID- 9328937 TI - Ischemia causes rapidly progressive nephropathy in the diabetic rat. AB - We examined the influence of renal ischemia in rats with diabetes mellitus (DM). Male Wistar rats were rendered diabetic by streptozotocin treatment. Two weeks later, 30 minutes of complete ischemia was induced in the left kidney of DM and non-DM animals. Both groups were evaluated functionally and morphologically four or eight weeks post-ischemia. In non-DM animals renal function and morphology showed almost complete recovery. In the DM animals, however, this comparatively short period of ischemia caused a substantial loss of renal function. Four weeks post-ischemia inulin clearance in the DM kidneys rendered ischemic was only 20% of that in the corresponding non-DM kidneys, and after eight weeks the DM kidneys were completely anuric. Extensive inflammation and tubulointerstitial fibrosis were evident in DM kidneys four weeks after ischemia and seemed to increase over time. After eight weeks, tubular atrophy was found in the ischemic DM kidneys, resulting in a substantial loss of kidney mass. We conclude that in diabetic rats renal ischemia causes rapidly progressive kidney damage that in several respects resembles diabetic nephropathy in humans. Since advanced renal lesions similar to those seen in human diabetic nephropathy never develop in the rat solely as a result of DM, the present study may provide an experimental model for further studies on renal failure in diabetes mellitus. PMID- 9328938 TI - Adenovirus-mediated beta-galactosidase gene delivery to the liver leads to protein deposition in kidney glomeruli. AB - The many cell types of the kidney, precisely arranged, allow this organ to perform its complex physiologic functions. However, this architectural complexity makes gene transfer into the kidney difficult. One approach to delivering a therapeutic protein to the kidney is to transfer a gene to a non-renal tissue. Release of the protein into the circulation might then result in deposition in the kidney, if the protein has the appropriate molecular properties. In this study, we found that parenterally administered replication deficient adenovirus carrying the beta-galactosidase gene resulted in intense beta-galactosidase gene expression in hepatocytes. As a result of immune attack on transduced hepatocytes, beta-galactosidase protein from these cells is released into the circulation, transported, and deposited almost exclusively in kidney glomeruli. Intense beta-galactosidase activity was noted in both kidneys with a peak at two weeks following viral administration, concurrent with loss of beta-galactosidase positive hepatocytes. Consistent with our hypothesis of protein transfer, no beta galactosidase mRNA was detected in glomeruli. Moreover, systemically administered protein generated similar glomerular beta-galactosidase activity. Finally, co administration of murine CTLA4 Ig, an immunomodulator of T cell activation, with the adenovirus protected infected hepatocytes and markedly diminished glomerular beta-galactosidase activity. Collectively, these findings suggest that a therapeutic protein can be "targeted" to the renal glomerulus, utilizing the liver as a gene transfer organ. PMID- 9328939 TI - Renal biopsy collagen I mRNA predicts scarring in rabbit anti-GBM disease: comparison with conventional measures. AB - Progressive loss of normal structure associated with scarring is the hallmark of chronic diseases of most organs. To test the hypothesis that measurement of interstitial collagen mRNA levels would be a useful index to predict future scarring, we developed an assay to quantitate alpha 1(I) procollagen mRNA factored for GAPDH mRNA using RT-PCR (the "CI:G ratio"). We first defined conditions under which the assay could be used for analysis of renal biopsy samples. The CI:G ratio was then used to determine whether mRNA measurements performed at an early stage of inflammation (day 7) in a model of anti-GBM disease in the rabbit would predict outcome at day 30 as measured by interstitial and glomerular scarring and renal cortical hydroxyproline accumulation. The predictive value of this assay was compared to functional (serum creatinine and urine protein:creatinine ratio) and histologic (glomerular and interstitial scoring) parameters also measured at day 7. We found that the CI:G ratio alone provided a sensitive and discriminating assay over a wide range of renal injury that predicted various parameters of scarring with an average coefficient of determination (r2) of 0.69. This predictive power was higher than that found for conventional measures, which tended to have good discriminatory capacity over limited ranges of renal injury. The CI:G ratio provided significant additional predictive power over and above that available from combinations of conventional functional or histologic parameters. We conclude that measurement of the CI:G ratio in biopsy samples deserves further assessment as a potentially useful quantitative predictor of outcome that could lead to improved clinical decision making. PMID- 9328940 TI - Lovastatin-induced inhibition of renal epithelial tubular cell proliferation involves a p21ras activated, AP-1-dependent pathway. AB - Proliferation of tubular epithelial cells underlies the development of cystic lesions and the subsequent impairment of renal function after renal mass reduction. The effect of HMG CoA reductase inhibitors (HRI) on cell proliferation was investigated in rat renal proximal tubular epithelial cells in primary culture. Treatment of renal tubular epithelial cells with three different HRI reduced fetal calf serum (FCS)-induced [3H]-thymidine incorporation (IC50 values were 0.7 microM, 1.7 microM, and 1.6 microM for simvastatin, lovastatin, and compactin, respectively), and lovastatin blocked BrdUrd incorporation, as assessed by immunocytochemical studies. The proliferative effect of epidermal growth factor (EGF) was similarly abolished by lovastatin. The effect of lovastatin (1 microM) was prevented by 100 microM mevalonate, 5 microM farnesyl pyrophosphate and 5 microM geranylgeranyl-pyrophosphate (in percent of control value, 31% vs. 102%, 60%, and 82%, respectively) while cholesterol and other products of the mevalonate pathway were inactive. Immunoblot analysis showed that lovastatin decreased membrane-bound p21ras and inhibited FCS-induced c-fos and c jun protein expression. Furthermore, electrophoretic mobility shift assay demonstrated the functional impairement of AP-1 DNA binding activity in lovastatin-treated cells. In conclusion, these results demonstrate that HRI are antiproliferative in epithelial tubule cells and that this effect is exerted, at least in part, via inhibition of the p21ras-activated and AP-1 dependent mitogenic cascade. PMID- 9328941 TI - Urine and plasma levels of uroguanylin and its molecular forms in renal diseases. AB - Uroguanylin activates the intestinal and possibly the renal guanylate cyclase C receptor, and stimulates Cl- secretion. We developed a sensitive radioimmunoassay (RIA) for human uroguanylin and measured its concentration in the urine and plasma. Twenty-four-hour urinary excretion of immunoreactive (ir-) uroguanylin for persons with a high-salt diet (10 g/day) was 137.8 +/- 14.4 pmol/day, significantly higher than that for persons with a low-salt diet (7 g/day, 95.1 +/ 16.3 pmol/day, P < 0.05). There were significantly positive correlations between the urinary excretion of ir-uroguanylin and Na+, Cl-, K+ or cyclic GMP (cGMP). We demonstrated the presence of messenger RNA of guanylate cyclase C in the medulla of human kidney. The concentration of plasma ir-uroguanylin significantly correlated with that of serum creatinine (r = 0.71, P < 0.001). Biologically active uroguanylin-16 accounted for 99% of the endogenous uroguanylin molecules in normal urine and 60% in plasma, the remainder being the 10 kDa precursor. The precursor content increased in the urine and plasma as the severity of renal impairment increased. These findings suggest that bioactive uroguanylin-16 is involved in the regulation of electrolyte homeostasis and that the kidney participates in the metabolism and excretion of uroguanylin. PMID- 9328942 TI - Luminal arginine vasopressin stimulates Na(+)-H+ exchange and H(+)-ATPase in cortical distal tubule via V1 receptor. AB - Bicarbonate reabsorption was evaluated by stationary microperfusion of in vivo early distal (ED) and late distal (LD) segments of rat kidney. Intratubular pH was recorded by double-barreled H ion-exchange resin/reference (1 M KCl) microelectrodes for the determination of HCO3- reabsorption. In the presence of luminal arginine vasopressin (AVP, 10(-9) M), a significant increase in HCO3- reabsorption was observed both in ED (from 0.931 +/- 0.061 to 2.12 +/- 0.171 nmol.cm-2.s-1] and LD segments [from 0.542 +/- 0.086 to 1.67 +/- 0.111 nmol.cm 2.s-1]. The addition of the V1-receptor antagonist [(d (CH2)5, Tyr (Et)2) arginine vasopressin] (10(-5) M) to luminal perfusion blocked luminal AVP mediated stimulation in ED and LD segments. 5-(N, N-hexamethylene) amiloride (10( 4) M) added to luminal perfusion inhibited luminal AVP-mediated stimulation in ED (by 63.7%) and LD (by 34.1%) segments. The addition of Bafilomycin A1 (2 x 10(-7) M) to the luminal perfusion did not affect luminal AVP-mediated stimulation in ED segments, but reduced it (by 31.7%) in LD segments. Our results indicate that luminal AVP acts to stimulate the Na(+)-H+ exchange in ED and LD segments via activation of V1 receptors, as well as the vacuolar H(+)-ATPase in LD segments. PMID- 9328943 TI - Altered nitric oxide metabolism and increased oxygen free radical activity in lead-induced hypertension: effect of lazaroid therapy. AB - Chronic exposure to low levels of lead results in sustained hypertension (HTN) in humans and experimental animals. The mechanism of lead-induced HTN remains unclear. We investigated the possible role of reactive oxygen species (ROS) and their impact on nitric oxide (NO) metabolism in lead-induced HTN. Male Sprague Dawley rats were treated with lead (100 ppm in drinking water) for twelve weeks. They were then treated with either the potent antioxidant, lazaroid (des-methyl tirilazad, 5 mg/kg i.p., twice daily) (Pb-Lz group) or placebo (Pb group) for two weeks and monitored for an additional two weeks. A group of normal animals served as controls (N = 6 in each group). Lead administration resulted in marked HTN together with a significant rise in plasma concentration of lipid peroxidation product, malondialdehyde (MDA, reflecting increased ROS generation) and a twofold reduction in urinary excretion of NO metabolites, that is, total nitrates and nitrites (NOx). Lazaroid therapy led to prompt normalization of blood pressure, plasma MDA and urinary NOx. In contrast, blood pressure and plasma MDA remained elevated, and recovery of urinary NOx excretion was slow with placebo therapy. No significant difference was found in creatinine clearance between the study groups during the observation period. Thus, chronic lead exposure resulted in marked HTN coupled with increased ROS production and decreased urinary NOx excretion. Administration of the potent antioxidant, lazaroid, abrogated HTN and reversed the abnormalities of plasma MDA and urinary NOx excretion, thus supporting the role of ROS in lead-induced HTN in this model. PMID- 9328944 TI - Expression of the mucosal gamma delta T cell receptor V region repertoire in patients with IgA nephropathy. AB - IgA nephropathy (IgAN) is characterized by the deposition of IgA in the glomerular mesangium and often leads to progressive renal dysfunction and kidney failure. We have previously shown that the mesangial IgA is likely to derive from the bone marrow plasma cells, and suggested that a primary abnormality within the mucosal immune system may underly the pathogenesis of IgAN. This study has analyzed the T cell receptor (TCR) variable (V) region expression by gamma delta T cells in the intestinal mucosa of patients with IgAN. The V gamma and V delta usage of TCR transcripts was determined using a semiquantitative reverse transcriptase-PCR protocol. Primers specific for the four human V gamma and six V delta subfamilies were used each with a constant (C) gamma or C delta specific primer, and the PCR-amplified TCR transcripts were detected by Southern blotting and oligonucleotide hybridization. gamma delta TCR V region expression was determined in gut biopsies and peripheral blood of 11 patients with IgAN, and the TCR V gamma and V delta repertoires were compared to those in gut and peripheral blood of 11 control individuals. gamma delta T cells in normal blood predominantly expressed V gamma 2 (V gamma 9 gene) and V delta 2 gene segments whereas those in normal gut mainly expressed V gamma 3 and V delta 3. In IgAN patients, V delta 2 was also the predominant V delta gene utilized by peripheral blood gamma delta T cells, however, we observed a predominance of V gamma 3 and reduced V gamma 2 usage by these cells. gamma delta T cells in the gut of IgAN patients mainly used V gamma 3 and V delta 1. While the gamma and delta TCR V region repertoires did not differ significantly between the peripheral blood of patients and controls, there were significant differences in V gamma and V delta repertoire expression between IgAN and control gut biopsies. V gamma 3 gene expression was significantly decreased in IgAN gut compared to control gut (P = 0.023). In addition, there was a significant decrease in V delta 3 gene expression in IgAN gut compared to control gut (P = 0.043). These findings indicate that a subpopulation of gamma delta T cells, which represent the majority of gamma delta T cells in normal gut mucosa, are significantly diminished in the gut of patients with IgAN. This suggests that a "hole" in the mucosal gamma delta T cell repertoire may play a fundamental role in contributing to the pathogenesis of IgAN. PMID- 9328945 TI - In situ expression and soluble form of P-selectin in human glomerulonephritis. AB - To clarify the early involvement of cellular adhesion molecules in human glomerulonephritis, we investigated P-selectin and high endothelial venules' (HEVs) marker MECA-79 expression in kidney specimens by immunohistochemical and in situ hybridization analyses, and measured serum and urinary soluble P-selectin levels by enzyme-linked immunosorbent assay. In normal controls, P-selectin and MECA-79 expression were negative in glomeruli (N = 4), and serum soluble P selectin levels were 114.3 +/- 36.8 ng/ml (mean +/- SEM, N = 12). Soluble P selectin was not detectable in urine of all cases. In proliferative glomerulonephritis involving rapidly progressive glomerulonephritis (N = 6), IgA nephropathy (N = 26), lupus nephritis (N = 7) and acute glomerulonephritis (N = 2), both glomerular and interstitial P-selectin expression were up-regulated. Glomerular P-selectin expression correlated positively with local cellular accumulation, endocapillary proliferation and CD41b (platelet) staining. Interstitial P-selectin expression showed a positive correlation with the grade of local cellular infiltrates. P-selectin mRNA signals detected by in situ hybridization were only observed on capillary or venous endothelium in the interstitium, but not in glomeruli. In addition, MECA-79 was expressed on the plump endothelial cells at the cortico-medullary junction (outer medulla). Serum soluble P-selectin levels were significantly higher in patients with proliferative glomerulonephritis, especially in glomerular and interstitial P selectin positive staining, and correlated with glomerular endocapillary proliferation. These observations suggested that P-selectin was associated with both glomerular and interstitial leukocyte accumulation in human glomerulonephritis, and might be expressed by two distinct mechanisms that are the activated platelets in glomeruli and the de novo expression in the interstitial lesions that correlated with MECA-79 expression as HEVs like vessels, and serum soluble P-selectin may be a useful marker for predicting in situ P-selectin expression associated with glomerular endocapillary proliferation in nephritis. PMID- 9328946 TI - Technical note. The serum concentration of the advanced glycation end-product N epsilon-(carboxymethyl)lysine is increased in uremia. AB - Advanced glycation end products (AGEs) such as pentosidine and N epsilon (carboxymethyl)lysine (CML) have been traditionally quantified by HPLC or gas chromatography--mass spectrometry (GC/MS). Enzyme-linked immunosorbent assays (ELISA) have been introduced as a convenient alternative to simplify the detection and measurement of AGEs in proteins and tissues, but some of these studies are limited by the lack of information on the structure of the epitopes recognized by antibodies to AGE-proteins. In this work we demonstrate that an antibody used in a previous study, reporting increased levels of AGEs in patients with diabetes or on continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD), recognizes CML as its major epitope. We also show that there is a significant correlation between the concentration of AGEs in serum measured by ELISA and a GC/MS assay for CML in serum proteins. Both analyses yielded comparable results, with patients on CAPD and HD having about threefold higher AGE- or CML-concentrations in their serum. Our data suggest that ELISA assays for CML should be useful for the clinical measurement of AGEs in serum proteins. PMID- 9328947 TI - Effect of increased dialysate fill volume on peritoneal fluid and solute transport. AB - It has recently been recommended that the peritoneal dialysate volume should in general be increased to increase the peritoneal small solute clearances. However, the net ultrafiltration volume may decrease due to higher intraperitoneal hydrostatic pressure (IPP) and higher peritoneal fluid absorption induced by higher fill volume. In the present study, we investigated the effects of increasing the fill volume on peritoneal fluid and solute transport. A four-hour dwell study with frequent dialysate and blood sampling was performed in 32 male Sprague-Dawley rats using 16 ml, 25 ml, 30 ml or 40 ml (8 rats in each group) of 3.86% glucose solution with 131I albumin as an intraperitoneal volume marker. The peritoneal transport of fluid, glucose, urea, sodium, potassium, phosphate and total protein as well as IPP with different fill volume were evaluated. The IPP and peritoneal fluid absorption rate (as estimated from the 131I albumin elimination coefficient, KE) significantly increased with increase in fill volume (P < 0.05), whereas the direct lymphatic absorption did not change with increasing fill volume. There was a strong correlation between IPP and KE. However, the net ultrafiltration volume was significantly higher in the high fill volume groups compared to the low fill volume groups, mainly due to a better maintenance of the dialysate to plasma glucose concentration gradient in the high fill volume groups. There was no significant difference in the diffusive mass transport coefficients (KBD) and sieving coefficients for any of the investigated solutes, although KBD values tended to be lower in the 16 ml group. The clearances for small solutes increased with increased fill volume, although these increases were slightly smaller than predicted from the increase in fill volume. We conclude that: (1) An increase in dialysate fill volume using 3.86% glucose solution results in higher intraperitoneal hydrostatic pressure and higher peritoneal fluid absorption, but, on the other hand, a higher net ultrafiltration; (2) The increase in net ultrafiltration with increased fill volume is mainly due to a better maintenance of glucose concentration in the dialysate, inducing an increased transcapillary ultrafiltration rate; (3) Solute clearances increase although not quite to the same extent as predicted from the increase in fill volume. Our results indicate that decreased net ultrafiltration volume associated with higher dialysate fill volume (due to higher IPP and higher peritoneal fluid absorption) could be avoided if hypertonic glucose solutions are used. PMID- 9328948 TI - Beta 2-microglobulin associated amyloidosis: a vanishing complication of long term hemodialysis? AB - Beta 2-microglobulin associated amyloidosis (A beta 2m amyloidosis) is considered an inevitable complication of chronic hemodialysis, particularly in hemodialysis with cellulose based membranes. We performed a single center study to assess the prevalence of A beta 2m amyloidosis in 1988 versus 1996. Randomly selected patients, studied in 1988, were matched for time on hemodialysis (mean 71 months, range 3 to 207) and age (mean 51 years, range 22 to 80) with patients of the 1996 population. Compared to 1988 patients, the 1996 patients exhibited a lower prevalence of carpal tunnel syndrome (7 of 43 in 1988 vs. 1 of 43 in 1996; P < 0.001) and radiological evidence of A beta 2m amyloidosis (13 of 34 patients vs. 3 of 34 patients positive; P < 0.001; and 33 of 272 possible sites affected in 1988 vs. 7 of 272 sites in 1996 patients; P < 0.05). Compared to the 1988 population, the 1996 population exhibited significantly lower serum aluminum levels, lower average serum creatinine (but not urea) levels, more frequent therapy with erythropoietin, less home hemodialysis, longer hemodialysis time using high-flux synthetic dialysis membranes (mean of 13% vs. 6% of the total hemodialysis time in the 1988 group), and more frequent usage of reverse osmosis water plus bicarbonate buffer for dialysate preparation. We conclude that the prevalence and severity of A beta 2m amyloidosis unexpectedly decreased by about 80% in our center between 1988 and 1996. Given the relatively short times spent on high flux hemodialysis in both groups, increased beta 2-microglobulin removal is unlikely to account for this phenomenon. Rather, other factors, for example, dialysate composition and purity, may be involved. PMID- 9328949 TI - Pressure or flow recordings for the surveillance of hemodialysis grafts. AB - Venous pressures (VP) measured by the dialysis machine are widely used for access surveillance and have significantly improved outcomes. VP reflect the resistance in the venous outflow tract, which will rise in the presence of stenosis. Low graft flow caused by high graft resistance predicts thrombosis. In this study we investigated whether high VP coincides with low graft flow (measured by ultrasound dilution technique). Of 70 forearm bridge grafts in 42 chronic hemodialysis patients, 31 had an angiographically proven outflow stenosis. VP at 200 ml/min blood flow (VP200), total graft resistance and venous outflow resistance were higher whereas graft flow was lower in patients with venous outflow tract stenosis as compared to patients without stenosis. Diagnostic power of the tests for identifying patients with venous stenosis showed no important differences. However, arterial inflow resistance, which is not reflected in VP measurements, represented a substantial and, more importantly, a highly variable percentage of total graft resistance. As a result graft flow showed no correlation with VP measurements. In conclusion, although patients with venous outflow stenosis may be identified accurately using venous pressure assessments, graft flow measurements seem to be more suitable for selecting patients at risk for thrombosis. PMID- 9328950 TI - Role of an improvement in acid-base status and nutrition in CAPD patients. AB - Short-term correction of metabolic acidosis in normal and uremic subjects has been shown to decrease protein degradation, but the long-term effects of better correction of acidosis on nutrition in ESRF are unknown. The aim of this study was to assess the possible benefits, in the nutritional state and morbidity, of improved correction of acidosis in the first year of treatment with continuous ambulatory peritoneal dialysis (CAPD). Two hundred consecutive new CAPD patients were randomized, in a single-blind fashion, to receive a high (HA; lactate 40 mmol/liter) or low (LA; lactate 35 mmol/liter) alkali dialysate for one year. Calcium carbonate and sodium bicarbonate were also used to correct acidosis in the HA group. At one year, the venous serum bicarbonate and arterial pH were 7.44 +/- 0.004 and 27.2 +/- 0.3 mmol/liter in the HA group, and 23.0 +/- 0.3 mmol/liter and 7.4 +/- 0.004 in the LA group (P < 0.001). Dialysis dose, at one year or at the point of leaving the study (HA 8.0 +/- 0.1 liters/day vs. LA 8.5 +/- 0.3 liters/day) was not significantly different (P = 0.18). At one year, the increase in body weight in the HA group (6.1 +/- 0.66 kg) was higher than in the LA group (3.71 +/- 0.56 kg, P < 0.05). The increase in midarm circumference in the HA patients (1.26 +/- 0.16 cm) was significantly higher than the increase in the LA patients (0.61 +/- 0.16 cm, P < 0.05). The increase in triceps skinfold thickness were not significantly different (HA 2.5 +/- 0.41 mm vs. LA 1.24 +/- 0.38 mm, P = 0.1). Serum albumin was 37.8 +/- 0.4 g/dl at one year in the HA group, and 38.2 +/- 0.5 g/dl in the LA group (NS). Dietary protein intake at one year (HA 0.9 +/- 0.2 g/kg/day vs. LA 1.0 +/- 0.1 g/kg/day) was not significantly different. There were fewer hospital admissions in the HA group (1.13 +/- 0.16 per patient per year) compared to the LA group (1.71 +/- 0.22 per patient per year, P < 0.05). The HA patients spent less days in hospital per year than the LA patients (16.4 +/- 1.4 days/year vs. 21.2 +/- 1.9 days/year; P < 0.05). It is concluded that better correction of metabolic acidosis leads to greater increases in body weight and midarm circumference, but not triceps skinfold thickness, in the first year of CAPD. The improvement in morbidity, in terms of number of admissions and days in hospital per year, may be associated with the improvement in nutritional state. PMID- 9328952 TI - Biology of acute renal failure: therapeutic implications. PMID- 9328951 TI - Switch from conventional to high-flux membrane reduces the risk of carpal tunnel syndrome and mortality of hemodialysis patients. AB - The use of a high-flux membrane, which eliminates larger molecular weight solutes with better biocompatibility, has steadily increased since the discovery of beta 2 microglobulin (beta 2m) amyloidosis in 1985. The long-term effects of a dialyzer membrane on morbidity and mortality are not completely understood. To examine the membrane effect as a factor of carpal tunnel syndrome onset and mortality, multivariate Cox regression analysis with time-dependent covariate was conducted on 819 patients from March 1968 to November 1994 at a single center. Two hundred and forty-eight of the patients were either switched from the conventional to high-flux membrane or treated only with a high-flux membrane. Fifty-one patients underwent a CTS operation and 206 died. Membrane status (on high-flux or on conventional) was considered as time-dependent covariate and risk was adjusted for age, gender, type of renal disease and calendar year of dialysis initiation. The relative risk of CTS was reduced to 0.503 (P < 0.05) and mortality 0.613 (P < 0.05) by dialysis on the high-flux membrane, compared to the conventional membrane. Serial measurements of beta 2m indicated significantly lower beta 2m to persist in patients on the high-flux membrane. The high-flux membrane decreased the risk of morbidity and mortality substantially. Larger molecule elimination was shown important not only for preventing beta 2m amyloidosis, but for prolonging survival of dialysis patients as well. PMID- 9328953 TI - Protein kinases in the action of vasopressin. PMID- 9328954 TI - Expression and subcellular localization of water channel aquaporin-2 in conscious rats. PMID- 9328955 TI - The Wistar-Furth rat as a model of mineralocorticoid resistance. PMID- 9328956 TI - Interaction of kinins and captopril in regulating arterial smooth muscle cell proliferation. PMID- 9328957 TI - Role of receptor mediated endocytosis in proximal tubule epithelial function. PMID- 9328958 TI - Central endothelin 1 regulation of arterial pressure and arginine vasopressin secretion via the AV3V region. PMID- 9328959 TI - Cellular mechanisms of cyclosporine A-associated side-effects: role of endothelin. AB - Immunosuppressive therapy with cyclosporine A (CsA) may be associated with severe side-effects such as nephrotoxicity and arterial hypertension. The partial reversability of these effects suggests that they are at least in part functional. We examined the effects of CsA on cellular signaling in cultured vascular smooth muscle cells from rat aorta. Intracellular free calcium concentrations ([Ca2+]i) were measured using fura-2. Total cell calcium was measured by atomic absorption and cellular endothelin production was estimated by radioimmunoassay. In the presence of CsA the calcium mobilizing effect of angiotensin (Ang) II was significantly enhanced. While the ETA receptor antagonist BQ 123 did not affect Ang II-induced calcium mobilization, the potentiating effect of CsA on [Ca2+]i was blocked by BQ 123. Preincubation of the cells with cyclosporine (10 micrograms/ml) for 30 minutes increased total cell calcium from 2.6 +/- 0.5 to 6.9 +/- 0.3 nmol/mg protein (P < 0.01). Within 24 hours endothelin production was significantly enhanced in the presence of cyclosporine (52.2 +/- 2.5 vs. 65.9 +/- 2.7 fmol/mg protein, P < 0.05). Therefore, the cyclosporine-induced rise of total cell calcium in smooth muscle cells is associated with an enhanced production of endothelin. We speculate that cyclosporine induced changes of Ca(2+)-kinetics may be mediated by endothelin. These results indicate that endothelin may play a major role in cyclosporine associated side-effects. PMID- 9328960 TI - Role of glycosphingolipids in the regulation of renal phosphate transport. PMID- 9328961 TI - Growth factor regulation of renal tubular epithelial cell motility. PMID- 9328962 TI - Nitric oxide in acute renal failure: foe or friend? PMID- 9328963 TI - Vascular hyporesponsiveness in cirrhotic rats: role of different nitric oxide synthase isoforms. PMID- 9328965 TI - Pathogenesis of cardiac dysfunction during metabolic acidosis: therapeutic implications. AB - Based on work performed in many laboratories including our own, we suggest the following schematic shown in Figure 4 to explain metabolic acidosis and the effects of alkalinization therapy. Metabolic acidosis induces prompt and substantial decreases in cardiac functional performance. This is mediated by an intracellular acidosis which impairs cardiac function directly as well as leads to an impairment of cardiac energy metabolism which further impairs cardiac function. Attempts to alkalinize the cell with sodium bicarbonate therapy lead to a paradoxical intracellular acidosis and further impairment of cardiac function, whereas Carbicarb may correct the intracellular acidosis and improve physiological function. However, at the time which this paper was written, Carbicarb was not available for clinical use in the United States. PMID- 9328964 TI - The glomerular cell: source and target of various mediators. PMID- 9328967 TI - Role of endothelin-1 in the activation of polymorphonuclear leukocytes. PMID- 9328966 TI - Integrins and repair after acute renal injury. PMID- 9328968 TI - Proteinuria and tubulointerstitial injury. AB - These studies have demonstrated pathways whereby one urinary protein, holotransferrin, may alter proximal tubular cell function and cause tubular cytotoxicity, and at least two urinary proteins, albumin and transferrin, may mediate the development of interstitial inflammation in proteinuric renal disease. PMID- 9328969 TI - Efficacy of immunosuppressive treatment in patients with membranous nephropathy and renal insufficiency. PMID- 9328970 TI - Primary IgA nephropathy: natural history and factors of importance in the progression of renal impairment. PMID- 9328971 TI - Hypertension in autosomal dominant polycystic kidney disease. PMID- 9328972 TI - Hydrochlorothiazide, atenolol, nitrendipine, and enalapril in antihypertensive treatment. Influence on LVH, proteinuria and metabolic parameters. The HANE Trial Research Group. PMID- 9328973 TI - Prognostic significance of renal dysfunction in cirrhosis. PMID- 9328974 TI - Non-insulin dependent diabetes mellitus in nephrology: where do we go from here? PMID- 9328975 TI - Gene therapy for renal diseases. AB - Gene therapy is a promising therapeutic approach for a variety of renal diseases including both inherited and acquired diseases. In vivo gene transfer in the kidney using viral or non-viral vectors have been reported. These approaches have been tested in a few animal models of renal diseases, including experimental glomerulonephritis, ischemic renal failure, and carbonic anhydrase II deficiency. Selection of vectors, routes, and therapeutic genes is critical to the success of gene therapy targeted to the specific compartment of the kidney. Limitations of gene therapy for renal diseases exist and consist of: duration of transgene expression is short, transfection efficiency is not adequate, immune reactions are induced by adenoviral vector, and insertional mutagenesis may be caused by retroviral and adeno-associated viral vectors. Further studies are needed for improvement of gene delivery, minimization of side effects and development of cell-specific and long-term regulated gene expression. PMID- 9328976 TI - Total lymphoid irradiation for pretransplant immunosuppression: nine year follow up. AB - Total lymphoid irradiation (16 to 30 Gray units) was given to 12 patients (8 who had rejected previous grafts). We found a fall in T and B cell counts with a reversal of CD4/CD8, and a decrease in mitogeneic and allogeneic responses with recovery to pretransplant levels in 24 to 36 months. The total lymphoid irradiation patients (on low dose prednisone and cyclosporine) had nine year graft and patient survival rates of 50% and 81.5%, respectively, versus 56% and 69.8% for 65 patients on conventional immunosuppression. Complications included viral (7) and fungal (1) infections, immunological thrombocytopenia (3), hypothyroidism and radiation pneumonitis and pericarditis (1 each), and there were no non-skin malignancies. There was only one episode of acute rejection in the group with total lymphoid irradiation as compared to 18 in the other group. We believe that total lymphoid irradiation provides effective immunosuppression and has the potential for reducing and possibly eliminating other immunosuppressants. PMID- 9328977 TI - Treatment of sebaceous gland hyperplasia with the pulsed dye laser. AB - BACKGROUND AND OBJECTIVE: Sebaceous gland hyperplasia may be treated by cryotherapy, cauterization, topical chemicals, or excision. The major disadvantage of these therapeutic strategies is a considerable risk of postoperative scarring or dyspigmentation. The pulsed dye laser may be an effective and safe alternative treatment option. STUDY DESIGN AND METHODS: Our report presents two patients with sebaceous gland hyperplasia who were treated with the pulsed dye-laser (585 nm, 6.5-8 J/cm2, 300-450/microsecond). RESULTS: After 2-3 treatment sessions, the lesions were completely gone. To data, no side effects have been observed. CONCLUSIONS: Based upon our experiences, we recommend the pulsed dye laser as a safe, fast, and minimal straining treatment alternative for hyperplasia of sebaceous glands. PMID- 9328978 TI - Photodynamic therapy for Barrett's esophagus: cardiac effects. AB - BACKGROUND AND OBJECTIVE: Atrial fibrillation has been reported following esophageal photodynamic therapy. This study presents the results of serial cardiac testing following photodynamic therapy for patients with Barrett's esophagus and with dysplasia or early carcinoma. STUDY DESIGN/MATERIALS AND METHODS: Twelve patients were treated using photodynamic therapy. Serum creatinine phosphokinase and lactic dehydrogenase isoenzyme levels were determined pretreatment and 24, 48, and 72 hours after treatment. Electrocardiograms were obtained before and 48 hours after treatment. A rhythm strip was obtained 1 week posttreatment. Clinical assessment for cardiac arrhythmias occurred daily following therapy. RESULTS: Transient atrial fibrillation was noted in one patient during a follow-up endoscopy. However, no significant or permanent abnormality was noted in cardiac enzymes or electrocardiograms. CONCLUSION: No permanent electrocardiographic changes or significant abnormalities in cardiac enzymes were detected following esophageal photodynamic therapy in patients with or without histories of cardiac disease. Delivery of esophageal PDT is not associated with permanent adverse cardiac effects. PMID- 9328980 TI - Copper bromide laser treatment of facial telangiectasia: results of patients treated over five years. AB - BACKGROUND AND OBJECTIVE: Various yellow light lasers have been used over the past decade in an attempt to eradicate facial telangiectasia. Based on their power output, spot size, and pulsing characteristics, these lasers belong to one of two categories that exist at either end of a spectrum--high power, short pulse, and large spot size, or low power, long exposure, and small spot size. The copper bromide laser clearly belongs in the latter group, but with higher available power than most other lasers in this group, it exists further along the spectrum toward the region in which the laser parameters might be considered closer to theoretical ideals for treating certain cutaneous vascular pathologies. The objective of this study was to ascertain the role and efficacy of the copper bromide laser on treatment of a variety of facial telangiectasia. STUDY DESIGN/MATERIALS AND METHODS: A total of 570 patients with facial telangiectasia of different diameters and on different regions of the face were treated with the copper bromide laser one or more times and followed up over 5 years. RESULTS: More than 75% clearance was achieved in 70% patients, 50-75% clearance in 17.4% patients, and < 50% clearance in 12.6% patients. Poor results were correlated with anatomical location on the nasal alae and nasal tip and also with vessel size. Very small (< 100 microns) and very large (> 300 microns) vessels did not respond as well as vessels in the 100-300-micron diameter group. Very large vessels responded better to a combination of sclerotherapy and laser treatment. There were no reported long-term adverse effects. CONCLUSION: The copper bromide laser is a safe and effective modality for the treatment of the majority of facial telangiectasia. It is less suited to treating very small vessel lesions such as diffuse erythema, and conversely very large vessels as well as those of the nasal alae. These latter two groups respond better and more permanently to combined sclerotherapy and laser treatment. PMID- 9328979 TI - Transurethral balloon laser enhanced thermotherapy in the canine prostate. AB - BACKGROUND AND OBJECTIVE: Hyperthermia is performed for prostate cancer. We examined the selective induction of coagulonecrotic changes in the objective area of the canine prostate in enhancing the effect of hyperthermia and treating the target area with transurethral balloon laser enhanced thermotherapy (TUBAL-ET) using a light absorbent material. STUDY DESIGN/MATERIALS AND METHODS: The heat exchange of ultrafine carbon particles after laser irradiation was observed in a phantom study using thermography. The carbon solution was injected at the right prostatic lobe in dogs and TUBAL-ET was performed. RESULTS: The charcoal absorbed the Nd:YAG laser energy and apparently converted it into thermal energy in the phantom study by thermographic observation. TUBAL-ET induced coagulonecrotic changes only at the area at which carbon had been injected in the prostate gland. The necrotic tissue was almost absorbed at four weeks after treatment. CONCLUSIONS: TUBAL-ET induces tissue damage at the target area in the prostate gland. PMID- 9328981 TI - Effects of pulsed laser systems on stapes footplate. AB - BACKGROUND AND OBJECTIVE: The aim of the present study was to investigate the tissue ablation capacity of various pulsed lasers at the stapes footplate. STUDY DESIGN/MATERIALS AND METHODS: Isolated human stapes and bovine compact-bone platelets were used to determine the effective laser parameters and appropriate application technique for achieving a perforation measuring 500-600 microns in diameter. Of interest were also the shape and quality of the perforations, the reproducibility of the perforation effect, and the thermically altered marginal zones occurring at the footplate. Three pulsed laser systems were used: excimer, holmium:YAG (Ho:YAG), and erbium:YSGG (Er:YSGG) lasers. RESULTS: The tissue ablating effect of pulsed laser systems permits a precise and controlled management of the stapes footplate through low and readily reproducible ablation rates. The extent of thermic side effects at the footplate is lower in comparison to the purely thermically acting cw and superpulse laser systems. The Er:YSGG laser exhibits the highest ablation rate at the stapes and is thus the most effective laser for perforation of the stapes footplate. Though somewhat less effective, the Ho:YAG laser also appears to be suitable for stapedotomy. On the other hand, we do not consider the applied excimer laser (308 nm) to be particularly appropriate at the stapes because of its low ablation rates. CONCLUSION: Thus, the erbium laser could represent an alternative to the argon, KTP 532, and CO2 lasers, already clinically successful in stapes surgery. However, further studies are necessary to examine the transmission of thermic energy into the vestibule and the acoustic stress to the inner ear during laser stapedotomy, to be able to make a definitive statement about the safest and most effective laser system for stapes surgery. PMID- 9328982 TI - Treatment of viral infections with 5-aminolevulinic acid and light. AB - BACKGROUND AND OBJECTIVE: When 5-aminolevulinic acid (ALA) is exogenously supplied, protoporphyrin IX (PpIX) is accumulated in various cells and makes them light sensitive. The possibility of using such an approach for the treatment of viral infections was studied in this work. STUDY DESIGN/MATERIALS AND METHODS: ALA was added to cultured cells infected with human immunodeficiency virus (HIV). Accumulation of PpIX in the cells as well as virus infectivity after photodynamic treatment (PDT) were assessed. For in vivo studies, guinea pigs were infected with herpes simplex virus (HSV) and then administered ALA at intervals after infection. The animals were exposed to PDT at the site of infection 3 hours after ALA administration. Clinical observations and virus titration were made daily. For clinical studies, two patients with Molluscum contagiosum and Verrucae vulgares were treated with ALA fortified with an iron chelating agent and dimethylsulfoxide, followed 4 hours later by PDT. RESULTS: Cells that are infected with HIV accumulated PpIX upon addition of ALA in vitro. This accumulation was enhanced approximately two-fold in the presence of an iron chelator. Subsequent exposure to red light PDT drastically reduced the virus titer (> 99% for U1 cells latently infected with HIV). In guinea pigs infected with HSV, subsequent administration of ALA and exposure of the lesions to red light shortened the duration of vesicles' appearance from more than a week to a few days and reduced HSV titer in the lesions by > or = 5 log10. ALA-PDT treated AIDS patient suffering from Molluscum contagiosum or a kidney transplant patient with Verrucae vulgares showed greatly improved clinical symptoms one month after treatment. CONCLUSION: It is concluded that ALA-PDT could be effective in treating certain viral infections, particularly those resulting in warts. PMID- 9328983 TI - Photodynamic therapy for rheumatoid arthritis? AB - BACKGROUND AND OBJECTIVE: The only early surgical therapy of rheumatoid arthritis is synovectomy. But even an arthroscopic synovectomy is restricted to more or less big joints. It has been shown recently that for smaller joints a laser synovectomy is possible but more time-consuming than with mechanical instruments. An alternative method may be photodynamic therapy. STUDY DESIGN/MATERIALS AND METHODS: In this study, possible photodynamic effects of Chloroquine, Methotrexate, Piroxicam, and Sodium Morrhuate were examined using a cell culture model of human synovial fibroblasts from patients having rheumatoid arthritis. RESULTS: Incubation with Chloroquine or Methotrexate and subsequent laser irradiation at a wavelength of 351 nm resulted in an at least twenty-fold enhanced cytotoxicity. CONCLUSION: Both substances therefore may serve for a photodynamic therapy of rheumatoid arthritis. PMID- 9328984 TI - Selective laser photocoagulation of blood vessels in a hamster skin flap model using a specific ICG formulation. AB - BACKGROUND AND OBJECTIVE: The present study was undertaken to evaluate the selective laser photocoagulation of blood vessels in a hamster skin flap model using a specific indocyanine green (ICG) formulation. STUDY DESIGN/MATERIALS AND METHODS: Experiments were performed in a hamster skin flap model after injection of ICG in aqueous solution (ICGA), or after injection of a specific formulation of ICG (ICG in emulsion: ICGE). Laser irradiation was achieved 30 minutes after injection with a 300 microns fiber connected to a 805 nm diode laser (power = 0.8W, spot diameter = 1.3 mm and pulse exposure time lasting from 1 to 5 s). Macroscopic observation and acute histology were performed to compare the tissue effects obtained for each ICG formulation and to assess the selectivity of vessel damage. RESULTS: The ICGE clearance process was slowed down as compared to the ICGA process. After 30 minutes, the concentration of ICG in blood is higher (2.27 +/- 0.4, P < 0.003) for ICGE compared to ICGA. With ICGA, vessel coagulation required a minimum fluence of 240 J/cm2, which led to very significant skin damage. Conversely with ICGE, vessel coagulation required a fluence of 120 J/cm2. With such a fluence, no laser effect could be detected on the skin. Histological examination confirmed blood vessels coagulation in depth, whereas epidermis and dermis remained intact. CONCLUSION: The major restrictions of ICG in aqueous solution, which are the very-short half-life of ICG in blood and consequently the lack of selectivity in blood vessels after a few minutes, are alleviated when ICG is used in emulsion. ICG in emulsion increases the circulating half-life of ICG and moreover confines ICG in the vascular compartment. Thanks to this specific property, it is possible to obtain a selective vascular damage 30 minutes after injection. PMID- 9328985 TI - Short wave ultraviolet laser energy in porcine coronary arteries: medial cell death and neointimal formation. AB - BACKGROUND AND OBJECTIVE: Smooth muscle cell migration and proliferation from arterial media into the neointima are major factors in the restenosis process following coronary angioplasty. Because short wave ultraviolet (UV) radiation is cytotoxic for rat carotid artery smooth muscle cells, the aims of this study were to determine the effects of short wave UV irradiation on normal pig coronary arteries and to evaluate the efficacy of UV laser energy for reducing neointimal hyperplasia (NI). STUDY DESIGN/MATERIALS AND METHODS: In 13 pigs fed a normal diet, 37 coronary arteries were studied. UV laser light (275 nm) was applied in escalating doses from 0-16,353 mJ/cm2 via fiberoptic through a 20 mm PTCA balloon catheter. The pigs were euthanized at 21 days and histologic analysis performed. Arterial media was rendered acellular (ACM) in 20 of 33 irradiated coronary arteries (61%). The minimum UV energy density inducing ACM was 1348 mJ/cm2. The fraction of acellular media to internal elastic lamina length (ACM/IEL) was 0.79 +/- 0.29. RESULTS: No statistically significant difference was found between NI thickness at normal media sites (NM) vs. ACM sites (0.17 +/- 0.14 mm vs. 0.16 +/- 0.17 mm). No correlation was found between UV dose and NI formation (r = 0.307, P = 0.08). CONCLUSION: Short wave UV irradiation induces ACM in normal porcine coronary arteries. Induction of acellular media is not associated with a reduction of NI formation in this porcine coronary model. PMID- 9328986 TI - Laser ablation as a function of the primary absorber in dentin. AB - BACKGROUND AND OBJECTIVE: Infrared transmission spectra of dentin reveal a broad absorption band between 6.0 and 7.0 microns composed of absorption peaks of water, collagen and carbonated hydroxyapatite. The nearly constant absorption and the existence of absorption peaks of different tissue components were used to investigate ablation as a function of the primary absorber. STUDY DESIGN/MATERIALS AND METHODS: Laser ablation of dentin as a function of fluence was studied in the wavelength range between 6.0 and 7.5 microns using the Vanderbilt Free-Electron Laser (FEL). Depth and volume of the ablation crater were determined with a silicon replica method and subsequent confocal laser topometry. SEM investigations were performed on the irradiated surfaces. For the description of the experimental data an ablation model is developed. RESULTS: At all applied wavelengths we found a linear increase of ablation depth as a function of fluence above a threshold fluence. The lower absorption of dentin at 7.5 microns compared to the absorption at 6.0, 6.5 and 7.0 microns results in a greater ablation threshold. At 6.0, 6.5 and 7.0 microns wavelengths the ablation thresholds are comparable. The experimental data are in good agreement with an ablation model using a mean absorption coefficient of the target material. No thermal cracking is observed after ablation in dentin. The post ablative surface structure at 6.0 and 7.0 microns looks similar whereas at 7.5 microns the surface reveals a greater roughness. CONCLUSION: The ablation efficiency and threshold depend on the mean absorption but do not depend upon the chemical identity of the primary absorber in dentin. Calculations show that heat conduction during the laser pulse leads to a thermal equalization between the heated microstructures and surrounding tissue resulting in an ablation with little dependence on the primary absorber. PMID- 9328987 TI - Periodontal tissue regeneration in beagle dogs after laser therapy. AB - BACKGROUND AND OBJECTIVE: Class III periodontal furcations still represent a challenge for the periodontist. Aim of this study was to test the effect of CO2 laser on the treatment of class III furcation defects. STUDY DESIGN/MATERIALS AND METHOD: Class III furcation defects 3 mm deep were surgically induced on mandibular premolars on six male Beagle dogs, for a total of 36 defects. After 6 8 weeks of plaque accumulation, the mean depth was 6.8 mm. Quadrants were randomly assigned to a) CO2 laser therapy (laser), b) Guided Tissue Regeneration (GTR) procedure using Gore-Tex Membranes, (Gore Tex, Flagstaff, Arizona, USA) and c) Scaling and Root planing (Sc/Rp). CO2 laser beam (El.En, Florence, Italy) was applied to the root surfaces in defocused pulsed mode at 2W, 1 Hz and a duty cycle of 6%, and on periodontal soft tissues at 13W, 40 Hz, and a duty cycle of 40%. Control quadrants received either GTR procedure or Sc/Rp. Mechanical oral hygiene was provided. At 6 months the animals were sacrificed. RESULTS: The laser group showed new attachment formation averaging 1.9 mm (sd +/- 0.5), whereas GTR and Sc/Rp showed 0.2 mm (sd +/- 0.4) and 0.2 mm (sd +/- 0.5) respectively, being the differences statistically significant between the laser group and both GTR and Sc/Rp groups (p < 0.005). CONCLUSION: CO2 laser treatment of class III furcation induced formation of new periodontal ligament, cementum and bone. PMID- 9329009 TI - The cutaneous microcirculation: ultrastructure and microanatomical organization. AB - The cutaneous microcirculation is organized as two horizontal plexuses. One is situated 1-1.5 mm below the skin surface, and the other is at the dermal subcutaneous junction. Ascending arterioles and descending venules are paired as they connect the two plexuses. From the upper layer, arterial capillaries arise to form the dermal papillary loops that represent the nutritive component of the skin circulation. There are sphincter-like smooth muscle cells at the point where the ascending arterioles divide to form the arteriolar component of the upper horizontal plexus. At the dermal subcutaneous junction, there are collecting veins with 2-cusped valves that are oriented to prevent the retrograde flow of blood. Laser Doppler flowmetry (LDF) has demonstrated vasomotion of red cell flux localized to the sites of ascending arterioles. The simultaneous recording by LDF of red cell flux and the concentration of moving red blood cells from individual sites allows one to construct by computer topographic maps of these two valves. The two maps, based on initial studies using correlative skin biopsy specimens, can define 1-mm3 volumes of skin that are predominantly arteriolar in composition, predominantly venular in composition, or essentially devoid of all microvascular elements. The electron and light microscopic features that define the microvascular segments, when coupled with the ability of LDF to define the predominant microvascular segments under the probe, will allow one to study both the mechanisms of normal physiological states and the pathogenetic mechanisms underlying pathological skin disorders in which the microvasculature plays a predominant role. PMID- 9329010 TI - Redox control of ion channel activity in vascular endothelial cells by glutathione. AB - Oxidized glutathione (GSSG) is endogenously formed within vascular endothelial cells. The bioactivity of GSSG results in the oxidation of protein thiol groups, leading to changes in protein structure-function relationships. When ion channel protein thiols are the target of oxidation by GSSG, important changes in channel conductance, activity, and gating occur. In this review, we focus on two endothelial cell ion channels, the activities of which influence vascular cell signaling and the nitric oxide signaling pathway. The first channel is the GSSG operated cation channel that depolarizes the endothelial cell, leading to inhibition of capacitative Ca2+ entry. The second channel is the inositol 1,4,5 triphosphate (IP3)-operated Ca2+ channel that is responsible for the agonist stimulated release of Ca2+ from IP3-sensitive endoplasmic reticulum. GSSG acts to deplete IP3-sensitive Ca2+ stores, thereby attenuating the intracellular Ca2+ response to agonist stimulation. Together, these effects indicate that glutathione, which is formed endogenously within the cell, is a key physiological modulator of endothelial cell signaling. PMID- 9329011 TI - Attenuation of oxygen free radical formation and tissue injury during experimental inflammation by P-selectin blockade. AB - OBJECTIVE: Neutrophilic leukocyte rolling on postcapillary endothelium requires membrane interaction between SLex-containing ligands on neutrophils and P selectin on endothelial cells. The current sequence of studies was performed to explore the hypothesis that the leukocyte-endothelial rolling interaction not only precedes leukocyte migration but also is accompanied by oxygen free radical production and interstitial cell death in the rat mesentery. METHODS: The ratio of leukocyte rolling velocity on the endothelium of postcapillary venules and centerline red cell velocity was determined after topical application of platelet activating factor (PAF; 10(-8) mol/L). Superoxide formation was determined by an in situ nitroblue tetrazolium reduction to dark blue formazan crystals in the in situ mesentery preparation, and cell death was detected by nuclear staining with propidium iodide. RESULTS: Leukocyte rolling and subsequent adhesion was inhibited with a monoclonal antibody against P-selectin, PB1.3. Superoxide formation, as well as parenchymal cell death, was significantly enhanced in the mesentery after stimulation with platelet activating factor, and both could be significantly attenuated by reduction of the rolling leukocyte-endothelial interaction with PB1.3. CONCLUSIONS: These results provide direct evidence that the interaction of leukocytes and endothelium followed by migration of leukocytes into the interstitium is accompanied by enhanced oxidative stress and parenchymal cell death. Early interruption of the interaction provides significant protection even in the presence of a proinflammatory stimulus. PMID- 9329012 TI - Lung neutrophil retention and injury after intestinal ischemia/reperfusion. AB - OBJECTIVE: To define the mechanisms responsible for the lung leukosequestration and injury elicited by intestinal ischemia/reperfusion (I/R). METHODS: The effect of 120 minutes of superior mesenteric artery occlusion and 90 minutes of reperfusion on neutrophil deformability, lung neutrophil retention, and pulmonary microvascular permeability was determined. RESULTS: Compared with control surgery, intestinal I/R resulted in a significant increase in neutrophil stiffness (mean yield pressure [Pyield], 1.533 +/- 0.075 and 2.302 +/- 0.288 cm H2O, respectively) and lung neutrophil content (6.3 +/- 1.4 and 31.5 +/- 6.4 U/g wet weight, respectively). These changes were not affected by inhibition of neutrophil adherence before gut reperfusion. However, the increased lung microvascular permeability elicited by gut I/R (0.111 +/- 0.020 [control surgery] and 0.255 +/- 0.041 [I/R] mL/min/cm H2O/100 g lung tissue) was significantly attenuated by administration of antibodies directed against neutrophil or endothelial determinants of leukocyte adhesion. CONCLUSIONS: The results of this study suggest that intestinal I/R is a potent inflammatory stimulus that elicits an increase in neutrophil stiffness and lung neutrophil retention independent of neutrophil-endothelial cell adhesion. In contrast, the increased lung microvascular permeability elicited by gut I/R is attenuated by strategies that interfere with neutrophil-endothelial cell adhesion. PMID- 9329013 TI - Polymorphonuclear leukocytes released from the bone marrow preferentially sequester in lung microvessels. AB - OBJECTIVE: A hallmark of the systemic response to an inflammatory stimulus is the release of polymorphonuclear leukocytes (PMNs) from the bone marrow. This study was designed to measure the release of PMNs from the bone marrow and to determine their sequestration in the lung after an intravenous injection of either endotoxin (n = 5) or saline (n = 5). METHODS AND RESULTS: The thymidine analogue 5'-bromo-2-deoxyuridine (BrdU) was used to pulse label dividing PMNs in the bone marrow of rabbits (n = 13), and immunohistochemistry and morphometry were used to detect the release of BrdU-labeled PMNs into the circulation and to determine their sequestration in the lung. Endotoxin treatment caused a drop in the circulating PMN counts (3.3 +/- 0.08 at baseline to 0.12 +/- 0.02 x 10(9)/L at 1 hour after endotoxin), which was followed by a neutrophilia at 8 hours (6.3 +/- 1.1 x 10(9)/L, P < 0.01), an increase in circulating band cells (0.12 +/- 0.01 at baseline to 2.18 +/- 0.4 x 1(9)/L at 8 hours, p < 0.001), and an increase in the percentage of BrdU-labeled PMNs (0.01% +/- 0.004% at baseline to 26.1% +/- 3.2% at 8 hours, p < 0.001). Endotoxemia caused an arteriovenous difference in BrdU labeled PMNs across the lung (35.9% +/- 2.9% versus 26.1% +/- 3.1%, mixed venous versus arterial, p < 0.02). Morphometric studies showed that endotoxin caused sequestration of PMNs in the lung (2.2 +/- 0.4 versus 1.0 +/- 0.2 x 10(10), endotoxin versus saline, p < 0.03) with preferential retention of BrdU-labeled PMNs (0.79 +/- 0.21 versus 0.039 +/- 0.016 x 10(10), endotoxin versus saline, p < 0.05). The percentage of BrdU-labeled PMNs in the alveolocapillary walls was higher than in circulating blood (64.01% +/- 4.3% versus 26.1% +/- 3.2%, p < 0.01) in the endotoxin group. In vitro filtration of cells through 5-mm pore size filters showed that circulating BrdU-labeled PMNs, 8 hours after endotoxin, were preferentially retained in the filters (p < 0.01). CONCLUSIONS: We conclude that endotoxemia stimulates the bone marrow to release mature and immature PMNs. Compared to PMNs released from the bone marrow during normal turnover, these PMNs are less deformable and preferentially sequester in the lung microvessels. PMID- 9329014 TI - Effects of anastomoses on solute transcapillary exchange in countercurrent systems. AB - OBJECTIVE: To investigate effects of anastomoses between the descending vasa recta (DVR) and the ascending vasa recta (AVR) on the distribution of small solutes in the interstitial fluid of the renal medulla. METHODS: Countercurrent capillary loops, surrounded by a secretory epithelium, were used to model microvessels in the renal medulla. Anastomoses between the DVR and AVR were modeled as a decrease in cross-sectional area of the vessels and a decrease in flow velocity from the base to the tip of the capillary loop. When experimental data were used to evaluate parameters of the model, it was seen that diffusive transport of solute in the axial direction of the capillary was negligible, and the equations could be greatly simplified. RESULTS: General formulae of the solute concentration distribution were derived for different degrees of shunting between the two limbs of the capillary loop. Analytical solutions for the steady state solute distribution were obtained when the sizes of capillaries and flows in them were assumed to decrease linearly with the distance from the base to the tip of the capillary loop. When the effects of reduction in the size of the limbs were compared with the effects of reduction of flow velocities on solute distribution, it was found that, in the presence of anastomoses, change in flow velocity increases the axial gradient of the solute concentration more than change in the cross-sectional area. The combined effects of a decrease in flow velocity and cross-sectional area can easily double the axial gradient of the solute concentration for a modest degree of anastomotic shunting. CONCLUSIONS: In this study, we separated effects of anastomotic flow between the DVR and AVR from other factors affecting the complicated countercurrent solute exchanges in the renal medulla. Results from the model show that anastomoses increase the solute concentration in the medullary interstitium and also the axial gradient of the solute concentration there. PMID- 9329015 TI - Innervation of the gallbladder: structure, neurochemical coding, and physiological properties of guinea pig gallbladder ganglia. AB - The muscle and epithelial tissues of the gallbladder are regulated by a ganglionated plexus that lies within the wall of the organ. Although these ganglia are derived from the same set of precursor neural crest cells that colonize the gut, they exhibit structural, neurochemical and physiological characteristics that are distinct from the myenteric and submucous plexuses of the enteric nervous system. Structurally, the ganglionated plexus of the guinea pig gallbladder is comprised of small clusters of neurons that are located in the outer wall of the organ, between the serosa and underlying smooth muscle. The ganglia are encapsulated by a shell of fibroblasts and a basal lamina, and are devoid of collagen. Gallbladder neurons are rather simple in structure, consisting of a soma, a few short dendritic processes and one or two long axons. Results reported here indicate that all gallbladder neurons are probably cholinergic since they all express immunoreactivity for choline acetyltransferase. The majority of these neurons also express substance P, neuropeptide Y, and somatostatin, and a small remaining population of neurons express vasoactive intestinal peptide (VIP) immunoreactivity and NADPH-diaphorase enzymatic activity. We report here that NADPH-diaphorase activity, nitric oxide synthase immunoreactivity, and VIP immunoreactivity are expressed by the same neurons in the gallbladder. Physiological studies indicate that the ganglia of the gallbladder are the site of action of the following neurohumoral inputs: 1) all neurons receive nicotinic input from vagal preganglionic fibers; 2) norepinephrine released from sympathetic postganglionic fibers acts presynaptically on vagal terminals within gallbladder ganglia to decrease the release of acetylcholine from vagal terminals; 3) substance P and calcitonin gene related peptide, which are co-expressed in sensory fibers, cause prolonged depolarizations of gallbladder neurons that resemble slow EPSPs; and 4) cholecystokinin (CCK) acts presynaptically within gallbladder ganglia to increase the release of acetylcholine from vagal terminals. Results reported here indicate that hormonal CCK can readily access gallbladder ganglia, since there is no evidence for a blood-ganglionic barrier in the gallbladder. Taken together, these results indicate that gallbladder ganglia are not simple relay stations, but rather sites of complex modulatory interactions that ultimately influence the functions of muscle and epithelial cells in the organ. PMID- 9329016 TI - Fine structure of cholesterolosis in the human gallbladder and the mechanism of lipid accumulation. AB - Gallbladders with cholesterolosis removed surgically for cholelithiasis were studied by light and electron microscopy as well as by cytochemical methods to demonstrate the presence of free cholesterol in the epithelial cells. Lipid droplets were found not only in the submucosa, but also in the infranuclear cytoplasm of epithelial cells. These contained well developed mitochondria and an agranular endoplasmic reticulum. Macrophages were often present between the epithelial cells and the submucosa, and protruded numerous processes, which also contained well developed cell organelles, abundant lysosomes and lipid droplets. With the excessive lipid deposition, macrophages were filled with lipid droplets and became foam cells. In the epithelial cells, many reaction precipitates occurred after digitonin treatment and some of them were observed in the endoplasmic reticulum. It is suggested, therefore, that free cholesterol is absorbed by epithelial cells and thereafter becomes esterified in the endoplasmic reticulum and thus appears as lipid droplets. Lipid droplets synthesized in the epithelial cells may then be released into the intercellular space, and phagocytosed there by macrophages. It is thus suggested that macrophages filled with lipid droplets may become too large and rigid to pass through the endothelium of lymph vessels, and those large "foam cells" may cause the destruction of lymph vessels. Those sequential events should eventually advance the accumulation of foam cells in the submucosa. PMID- 9329017 TI - Ultrastructural aspects of human cystic duct epithelium as a result of cholelithiasis and cholesterolosis. AB - Although there is a large body of data on the gallbladder and the importance of the cystic duct in surgical procedures, there is insufficient data regarding the morphology of the human cystic duct. In the present study, transmission electron microscopic (TEM) and scanning electron microscopic (SEM) survey of several surgical and autopsy cystic ducts in cholelithiasis and cholesterolosis is reported. In cholelithiasis, similar to gallbladder epithelium, the cystic duct epithelial cells display minor-to-severe alterations of the epithelial surface accompanied by variable erosion of the epithelium. Areas of intact surface epithelium demonstrate microvilli-covered cells coated by a rich glycocalyx and mucous production. In other areas, apical excrescences are associated with mucus hyperproduction and secretory events. Lipoid bodies are also present in many cells and especially in many of the cells' subliminal apical areas. In cholesterolosis, mucous secretory granules appear dilated, fatty deposits are infrequent, and peculiar intracellular cholesterol deposits can be detected in the apical and subapical region of cells and around condensed mitochondria. Following elective cholecystectomies, predominantly in association with cholelithiasis, eroded areas were detected; therefore, it appears that the action of intraluminal calculi may be a principal causative factor in discrete epithelial erosions of the cystic duct. Intraluminal calculi/ debris, along with the alteration of mucus, cell sloughing, and a decreased pool of bile acids and motility may participate in the gallstone nucleation process. The peculiar cholesterol inclusions may also play a role in that nucleating process. PMID- 9329018 TI - Ground squirrel model for cholelithiasis: role of epithelial glycoproteins. AB - The cholesterol-fed Richardson's ground squirrel (Spermophilus richardsonii) has proven to be an effective animal model in which to study factors that influence cholesterol gallstone formation and associated alterations in the gallbladder epithelium. Ground squirrels of either sex, fed a 2% cholesterol-enriched diet, exhibit cholesterol monohydrate crystal precipitation within 24 hours and macroscopically visible cholesterol stones by 3 weeks. Data on bile chemistry, biliary cholesterol precipitation, and various mucosal alterations occurring prior to, during, and after stone formation were collected using sampling intervals from 6 hours to 20 weeks on the diet. The results indicate that mucin hypersecretion appears to be more closely related to the initiation of nucleation than does either bile calcium of pH. Mucus hypersecretion begins within 18 hours of diet initiation and continues throughout the 20 week experimental period. Apical excrescences became more common and were larger in size during the early stages of cholelithiasis. Administration of aspirin during the experimental period demonstrated an inhibition of mucin synthesis and release. Gallstones were not formed in these aspirin-treated animals. A lectin-binding panel for 10 epithelial glycoprotein-related sugars indicated the mucin secreted by the gallbladder epithelium of 7 day experimental animals differed from that of controls. The most obvious difference was the abolition of WGA binding in the experimental animals, suggesting an absence of sialic acid expression in the mucin during the lithogenic process. Ultrastructural histochemistry indicated that both sulphomucin and sialomucin were present in the secretory granules and within the surface mucus layer of both experimental and control animals. Experimental animals, however, exhibited a significant predominance for sulphomucin. This pattern varies from that typically seen in other regions of the gastrointestinal tract where sialomucins predominate during pathologic processes. PMID- 9329020 TI - Monolayer and three-dimensional cell culture and living tissue culture of gallbladder epithelium. AB - Several models for preparing and isolating human and animal gallbladder epithelial cells, including low-grade gallbladder carcinoma cells, as well as proposed systems for culturing these isolated epithelial cells are reviewed here. Several reports concerning tissue culture of the gallbladder are also reviewed. The cell culture systems are divided into monolayer cell culture on collagen coated or uncoated culture dishes or other culture substrate and three dimensional cell culture in collagen gel. To prepare and isolate gallbladder epithelial cells, digestion of the gallbladder mucosa, abrasion of the mucosal epithelial cells, and excision of epithelial outgrowth of mucosal explants are applied. In monolayer cell culture, most of the specific biological features of isolated and cultured cells characteristic to the gallbladder are gradually lost after several passages, though quantitative and objective analyses of the pathophysiology of cultured cells and their secretory substances can be performed. Tissue culture using explants of the gallbladder has mainly been used for physiological studies of the gallbladder, such as investigating the transport of water and electrolytes. In this tissue culture system, quantitative assessment is difficult, though the original and specific biological and histological characteristics of the gallbladder are retained. Three-dimensional collagen gel culture could be an ideal model combining monolayer cell culture and tissue culture systems, and create controllable conditions or environments when several biologically active substances, such as growth factors, proinflammatory cytokines and adhesion molecules, are added to the culture medium. Advantages and shortcomings of individual cultivation models are discussed, and selecting the culture model most appropriate to the purpose of the study will facilitate investigations of the biology and pathogenetic mechanisms of gallbladder diseases such as cholelithiasis. PMID- 9329019 TI - Cholelithiasis induced in the Syrian hamster: evidence for an intramucinous nucleating process and down regulation of cholesterol 7 alpha-hydroxylase (CYP7) gene by medroxyprogesterone. AB - This report reviews previously published studies from our laboratory and shows some recent morphological data obtained with scanning and transmission electron microscopy regarding gallstone formation and alteration of the gallbladder epithelium in the Syrian hamster model. Both male and female hamsters were treated with female sex steroids (estradiol alone, estradiol and medroxyprogesterone, medroxyprogesterone alone) during one month. The results show that the Syrian hamster is a good model to study bile changes, gallbladder structure changes, including gallstone formation, and the regulation of cholesterol metabolism at the molecular level. Arguments in favor of this animal model are presented and, during gallstone formation, epithelial cell changes, anionic mucus secretion, and formation of gallbladder luminal deposits can be demonstrated. Recent molecular biology observations related to the effect of female sex steroids on liver cholesterol 7 alpha-hydroxylase (CYP7) gene suggest that progestin alone or primed by estrogen down regulates CYP7 transcription and activity. In addition, progesterone in cell culture systems has been shown to enhance intracellular accumulation of free cholesterol by increasing its uptake and synthesis and by decreasing its esterification by inhibiting the activity of acylcoenzyme A: cholesterol acyltransferase. Non-esterified cholesterol is free to migrate to the extracellular spaces and may contribute to nucleation within the bile. It is suggested that these effects of progesterone on cholesterol metabolism combined with the CYP7 gene down regulation, physical changes in the mucus and the hypomotility of the gallbladder and biliary ducts result in hypersaturation of cholesterol in the bile which favors gallstone formation. PMID- 9329021 TI - Imaging supramolecular aggregates in bile models and human bile. AB - Investigation of cholesterol crystallization is essential for the understanding of gallstone formation. Previous work has revealed a variety of aggregates of different sizes and shapes prior to the appearance of "classical" plate-like cholesterol monohydrate crystals both in native biles and model systems. In this article, we review existing data based on various microscopic techniques and present data on microstructural pathways leading to cholesterol crystal formation in two different bile models and in native bile. In continuation of our recent investigation of microstructures in nucleating human bile, we now present data suggesting that polymorphism is not limited to complex native bile, but also appears in two, simplified model systems. These studies employed cryo transmission electron microscopy (cryo-TEM) and video-enhanced light microscopy, using Nomarski optics (VELM). Only the combined use of these two complementary, non-perturbing direct methods can cover the whole range of microstructures ranging from a few nanometers to several microns. Concentrated isotropic solutions of bile models, composed of cholesterol, lecithin and taurocholate, were diluted to induce cholesterol supersaturation and start an evolution of microstructures, leading to cholesterol crystallization. Initially, small spheroidal micelles were observed by cryo-TEM. Subsequently, uni-, oligo- and multilamellar vesicles, compatible with structures seen at the same time by VELM, appeared in coexistence with micelles. Thereafter, during a dynamic phase of cholesterol crystallization, filaments, tubular and helical microstructures, as well as classical plate-like cholesterol monohydrate crystals were noted by light microscopy. Eventually, large plate-like crystals were observed by VELM, while cryo-TEM revealed only small spheroidal micelles. The crystallization process in native human bile during ex vivo incubation was found to bear close resemblance to the findings in the model systems, further supporting the applicability of these systems to the exploration of microstructural aspects of nucleating human bile. PMID- 9329022 TI - Hypericin-induced apoptosis of human malignant glioma cells is light-dependent, independent of bcl-2 expression, and does not require wild-type p53. AB - Hypericin and tamoxifen are experimental agents for the adjuvant chemotherapy of malignant glioma. We report that hypericin and tamoxifen induce apoptosis of 7 human malignant glioma cell lines in a concentration- and time-dependent manner. Illumination is essential for the cytotoxicity of hypericin but not tamoxifen. Apoptosis is unaffected by inhibitors of RNA and protein synthesis or free radical scavengers, does not require wild-type p53 activity, and occurs in glioma cells expressing high levels of bcl-2. There is no correlation between hypericin and tamoxifen-induced cytotoxicity and inhibition of protein kinase C (PKC). Ectopic expression of a murine bcl-2 transgene provides modest protection from tamoxifen but does not affect hypericin toxicity. Hypericin and tamoxifen do not modulate glioma cell killing induced by tumor necrosis factor-alpha (TNF-alpha) or CD95 ligand. Both drugs augment the acute cytotoxicity of various cancer chemotherapy drugs but fail to shift their EC50 values in modified colony formation assays. These data do not provide further supportive evidence how to enhance the limited efficacy of tamoxifen treatment for human malignant glioma. However, hypericin is a promising agent for the treatment of malignant glioma if local photodynamic activation of hypericin in the glioma tissue can be achieved. PMID- 9329023 TI - Vascular dementia, hypertension, and the brain. AB - Ischemic vascular dementia is a clinical syndrome of acquired intellectual impairment with ischemic cerebral injury resulting from occlusion of cerebral blood vessels and loss of cerebral tissue caused by cerebrovascular disease. With increasing life expectancy, the developed countries have experienced a shift towards a progressively older population. As the average age of the population increases, the prevalence of cerebrovascular disease and vascular dementia is likely to increase. The risk of vascular dementia seems to be correlated with the epidemiologic risk factors of stroke, namely hypertension. Hypertension is thought to be directly associated with vascular dementia and preliminary evidence suggests an association between elevated blood pressure and impairments in cognitive functioning. Recent investigations have found significant associations between hypertension and cerebral dilation and left hemisphere atrophy, and an increased incidence of white matter hyperintensities among hypertensives. Treatment and prevention of vascular dementia and cognitive dysfunction in the elderly require attention to cerebrovascular risk factors, particularly hypertension. Vascular dementias are potentially preventable and cases of Alzheimer's disease with vascular components are becoming increasingly recognized. PMID- 9329024 TI - Temporal integration of pain from electrical stimulation of the skin. AB - The threshold of sensation and the threshold of pain in response to electrical stimulation (impulses of 1 msec duration) of the skin on the forearm or hand in individuals without pain were compared with the thresholds of individuals with chronic pain in the range 1 to 100 pulses per second repetition rate. The threshold of sensation in patients without pain was little affected by the repetition rate of the stimulation within the range studied, and the threshold for pain decreased exponentially with increasing repetition rate. In individuals with chronic pain the threshold of sensation was similar to that of individuals without pain over the entire range of stimulus repetition rates studied, but the threshold of pain in patients with pain was lower and less affected by the stimulus rate than it was in the individuals without pain, thus closer to the threshold of sensation. PMID- 9329025 TI - A nonlinear quasi-static model of intracranial aneurysms. AB - Biomathematical models of intracranial aneurysms can provide qualitative and quantitative information on stages of aneurysm development through elucidation of biophysical interactions and phenomena. However, most current aneurysm models, based on Laplace's law, are renditions of static, linearly elastic spheres. The primary goal of this study is to: 1. develop a nonlinear constitutive quasi static model and 2. derive an expression for the critical size/pressure of an aneurysm, with subsequent applications to clinical data. A constitutive model of an aneurysm, based on experimental data of tissue specimens available in the literature, was incorporated into a time-dependent set of equations describing the dynamic behavior of a saccular aneurysm in response to pulsatile blood flow. The set of differential equations was solved numerically, yielding mathematical expressions for aneurysm radius and pressure. This model was applied to clinical data obtained from 24 patients presenting with ruptured aneurysms. Aneurysm development and eventual rupture exhibited an inverse relationship between aneurysm size and blood pressure. In general, the model revealed that rupture becomes highly probable for an aneurysm diameter greater than 2.0 mm and a systemic blood pressure greater than 125 mmHg. However, an interesting observation was that the critical pressure demonstrated a minimal sensitivity to the critical radius, substantiating similar clinical and experimental observations that blood pressure was not correlated, to any degree, with aneurysm rupture. Undulations in the aneurysm wall, presented by irregular multilobulated morphologies, could play an important role in aneurysm rupture. However, due to the large variation in results, more extensive studies will be necessary for further evaluation and validation of this model. PMID- 9329026 TI - Experience with transcranial magnetic stimulation in evaluation of spinal cord injury. AB - Nine subjects (seven male, two female) underwent transcranial magnetic stimulation (TMS) toward the evaluation of spinal cord injury (SCI). The evaluation of SCI with TMS tended to support clinical findings. Those subjects with clinically complete injuries demonstrated no evoked muscle response below the level of injury. Those subjects with clinically incomplete injuries showed trends toward prolonged evoked muscle latencies on the weaker side. Facilitation tended to enhance distal muscle responses. With incomplete spinal injury, the facilitation maneuver allowed the recording of weak muscle responses as well as those otherwise not present at rest. Maximum anal sphincter contraction also helped facilitate muscle responses and tended to impart less noise in the recordings. Facilitation failed, however to produce a response in those subjects with clinically complete injuries. No subject experienced adverse effects during the study. TMS promises to be an effective tool for the evaluation of SCI. PMID- 9329028 TI - Symmetric temporal patterns in cortical spike trains during performance of a short-term memory task. AB - The trion model is a highly structured representation of cortical organization, which predicts families of symmetric spatial-temporal firing patterns inherent in cortical activity. The symmetries of these inherent firing patterns are used by the brain in short-term memory to perform higher level computations. In the present study, symmetric temporal patterns were searched for in spike trains recorded from cells in parietal cortex of a monkey performing a short-term memory task. A new method of analysis was used to map neuronal firing into sequences of integers representing relative levels of firing rate about the mean (i.e. -1, 0 and 1). The results of this analysis show families of patterns related by symmetry operations. These operations are: i. the interchanging of all the +1's and -1's in a given pattern sequence (C symmetry), ii. the inverting of the temporal sequence of the mapping (T symmetry), and iii. the combination of the two previous operations (CT symmetry). Patterns of a given family are found across cells, especially in the memory periods of the task; in most cases they reoccur within a given spike train. The pattern families predicted by the model and reported here should be further investigated in multiple microelectrode and EEG recordings. PMID- 9329027 TI - Enhanced cytokines delivery and intercellular adhesion molecule 1 (ICAM-1) expression in glioma by intracarotid infusion of bradykinin analog, RMP-7. AB - The effect of intracarotid infusion of the bradykinin analog, RMP-7, on blood-to tumor and blood-to-brain transport of three cytokines were investigated. Wistar rats with RG2 gliomas were utilized and a unidirectional transfer constant, Ki, was determined using quantitative autoradiography. Interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and interleukin-2 (IL-2) were labeled with 125Iodine for quantitative transport studies using autoradiography. Radiolabeled cytokines were injected as an intravenous bolus. Intracarotid infusion of RMP-7 (0.1 microgram kg-1 min-1) increased the selective transport to tumors of IFN-gamma by 3.97-fold (p < 0.005), of TNF-alpha by 5.30-fold (p < 0.005), and of IL-2 by 4.34-fold (p < 0.005), compared to intracarotid saline infusion. To determine whether the increased IFN-gamma or TNF-alpha transport to tumors with RMP-7 could enhance expression of intercellular adhesion molecule 1 (ICAM-1) in tumors, ICAM-1 expression in RG2 glioma was evaluated by immunohistochemistry. Both IFN-gamma and TNF-alpha increased ICAM-1 expression of RG2 cells in vitro. In vivo, intracarotid infusion of IFN-gamma combined with RMP 7 significantly enhanced ICAM-1 expression in intracerebral RG2 gliomas compared to infusion of IFN-gamma without RMP-7. Expression of ICAM-1 was not enhanced by TNF-alpha combined with RMP-7. Intracarotid infusion of RMP-7 is a novel method of cytokines delivery to brain tumors. PMID- 9329029 TI - Intraventricular atrial natriuretic peptide for acute intracranial hypertension. AB - The effect of intraventricular atrial natriuretic peptide (ANP) on elevated intracranial pressure (ICP) was evaluated in a rodent model of global ischemia and reperfusion. ANP administration into the lateral ventricle 30 minutes after reperfusion statistically significantly reduced ICP compared with both vehicle treated animals (p < 0.001) and pretreatment pressures (p < 0.001). The ICP effects of ANP did not coincide with specific changes in regional perfusion parameters measured by laser-Doppler flowmetry. Two different vehicles for ANP were used to verify that the changes in ICP observed were independent of the sodium content administered in the vehicle. Based on the reductions observed in ICP, ANP deserves further evaluation as a treatment modality for the acute elevations in ICP associated with ischemic brain injury. PMID- 9329030 TI - Mediation of endothelium-dependent relaxation: different response patterns in rat and rabbit basilar artery. AB - The endothelial lining of blood vessels plays an important role in the control of arterial tone by releasing a number of vasoactive factors. Among these, nitric oxide (NO) or a NO containing moiety, some cyclooxygenase products such as prostacyclin (PGl2) and a still hypothetical hyperpolarizing factor may induce dilatation/relaxation. In the present study, the functional importance of these factors in mediating endothelium-dependent relaxation has been investigated in isolated cerebral arteries. Acetylcholine (ACh) was used to induce endothelium dependent relaxation of ring segments obtained from rat and rabbit basilar artery (BA). Glibenclamide (10(-6) M) which acts as an inhibitor of ATP-sensitive K+ channel activation did not affect the relaxant action of ACh. With the cyclooxygenase blocked by indomethacin (10(-6) M) ACh-induced relaxation was unaffected in rat BA and only slightly attenuated in rabbit BA. The NO synthase inhibitor NG-nitro-L-arginine (L-NNA, 10(-6) and 10(-5) M) completely abolished ACh-induced relaxation in rat BA and resulted in a moderate inhibition in rabbit BA (10(-5) and 10(-4) M). However, ACh-induced relaxation of rabbit BA was abolished in the presence of both indomethacin and L-NNA. These results indicate that endothelium-dependent relaxation of large cerebral arteries is mediated by NO acting alone or in concert with a relaxant cyclooxygenase product. In addition, there is no indication for the action of an endothelium-derived hyperpolarizing factor different from NO or a prostanoid. PMID- 9329031 TI - HEPES inhibits contractile responses of canine basilar artery. AB - N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid (HEPES) is a commonly-used buffer. This study determined the effect of HEPES on contractility of the dog basilar artery and tested the hypothesis that HEPES inhibits vasoconstriction of isolated arterial segments by generating H2O2. Rings of dog basilar artery with or without endothelium were suspended under isometric tension and contracted with KCl, serotonin, or prostaglandin F2 alpha (PGF2 alpha) in bicarbonate or HEPES buffer. Addition of HEPES, 30 mmol l-1, before or after contraction with KCl, serotonin or PGF2 alpha significantly decreased maximal tension in rings with or without endothelium. Preincubation with HEPES buffer, 10 mmol l-1, significantly decreased maximal contractions to each agonist in rings with endothelium and to KCl and serotonin in rings without endothelium. HEPES, 30 mmol l-1, noncompetitively inhibited concentration-contraction curves to increasing concentrations of each agonist in rings with or without endothelium. Inhibition by HEPES was completely reversible with washing. The inhibitory effects of HEPES on responses to each agonist in rings with endothelium were significantly less in the dark or after coincubation with catalase. Unlike HEPES, effects of H2O2 were endothelium-dependent in that H2O2 caused contractions in rings with endothelium and relaxations in rings without endothelium. 5-(N,N'-dimethyl)-amiloride and 4,4'-diisothiocyanataostilbene-2,2'-disulfonic acid did not affect contractility in this preparation. These results show that HEPES exerts significant inhibitory effects on arterial smooth muscle contractility. The mechanism does not involve endothelium-dependent relaxation, effects on chloride channels or the sodium hydrogen exchanger or generation of H2O2 by HEPES in the light. PMID- 9329032 TI - beta-Amyloid induces cerebrovascular endothelial dysfunction in the rat brain. AB - beta-Amyloid toxicity plays a central role in the pathology of Alzheimer's disease. Contraction and relaxation responses of pressurized rat posterior cerebral artery were studied before and after in vitro exposure to beta-amyloid. The peptide-induced characteristic features of endothelial dysfunction including enhanced vasoconstriction with serotonin and diminished relaxation to endothelium dependent vasodilators acetylcholine and bradykinin. Response to the endothelium independent vasodilator nitroprusside was not affected by beta-Amyloid. beta amyloid inhibition of acetylcholine-induced vasodilation was prevented by the oxygen radical scavenging enzyme superoxide dismutase. Endothelial destruction and the protective effect of superoxide dismutase was verified by electron microscopy. The results suggest that beta-amyloid peptide produces endothelial dysfunction in cerebral microvessels through reactive oxygen species. PMID- 9329033 TI - Effects of topical N-methyl-D-aspartate on blood-brain barrier permeability in the cerebral cortex of normotensive and hypertensive rats. AB - This study was performed to examine if blood-brain barrier (BBB) permeability could be increased by N-methyl-D-aspartate (NMDA) in the cerebral cortex, and to compare the degree of alteration of BBB permeability in normotensive and in chronic hypertensive rats. Twenty- to 22-week-old spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were anesthetized with isoflurane. After craniotomy in 7 animals in each group (SHR and WKY group), an NMDA patch (10 mM) was placed on one cortex (ipsilateral cortex: IC), and a normal saline patch was placed on the other cortex (control cortex: CC). The other 7 rats in each group were pretreated with MK-801 before placing NMDA and normal saline patches (SHR.MK 801 and WKY.MK-801 group). The BBB transfer coefficient (Ki) was determined using 14C-alpha-aminoisobutyric acid. The mean arterial pressures of the SHR and the SHR.MK-801 group were about 65% higher than those of the WKY and the WKY.MK-801 groups. In the WKY group, the Ki of the IC was significantly higher than that of the CC (IC: 10.0 +/- 2.7, CC: 6.2 +/- 2.4 microliters g-1 min-1). In the WKY.MK 801 group, the Ki was similar in both cortices (IC: 8.6 +/- 4.0, CC: 8.2 +/- 3.3). In the SHR group, the Ki of the IC was significantly higher than that of the CC (IC: 9.5 +/- 3.7, CC: 6.5 +/- 3.4), and the Ki of each cortex was similar to that of the corresponding cortex of the WKY group. In the SHR.MK-801 group, the Ki was similar in both cortices (IC: 7.2 +/- 1.5, CC: 7.1 +/- 2.7), and was also similar to those of the WKY.MK-801 group. Our data suggest that NMDA is involved in increasing BBB permeability. In chronic hypertension, the response of the BBB to NMDA is not altered when compared with normotension. PMID- 9329034 TI - A new nonNMDA antagonist modifies the local cerebral glucose utilization in kainate-induced generalized seizure. AB - To investigate the effects of a new nonNMDA antagonist on the trisynaptic pathways in the hippocampus, the author examined kainate(KA)-induced generalized seizures in rats. A novel nonNMDA antagonist, YM90K, showed the blockade of the Schaffer collaterals in 2-deoxyglucose study (2-DG) and that the CA1-2 pyramidal cells of the hippocampus were preserved seven days after the KA injections. On the other hand, the control and MK-801 (NMDA-antagonist) treated rats did not depress the Schaffer collaterals and showed persistent hypermetabolism of glucose in the CA1 pyramidal cell layer, where neurons were not preserved seven days later. 2-DG was useful to reveal the effects of nonNMDA antagonist on the KA induced generalized seizures. This suggests that YM90K is a potent nonNMDA antagonist and that it has a neuroprotective effect in rats. PMID- 9329035 TI - Pentoxifylline suppression of TNF-alpha mediated axonal degeneration in the rabbit optic nerve. AB - In AIDS patients, axonal degeneration in the optic nerve occurs as a histopathological manifestation of the optic neuropathy. Direct infection of neurons by HIV is unlikely, and the axonal injury may be an indirect effect mediated by cytotoxic factors such as tumor necrosis factor-alpha (TNF-alpha) which we have previously demonstrated to cause axonal degeneration in the rabbit optic nerve. To test the suppressive effects of pentoxifylline in preventing TNF alpha-mediated axonal degeneration, we applied pentoxifylline to an established rabbit model that demonstrates an AIDS-like optic neuropathy using intravitreal TNF-alpha injections. Degenerated axonal profiles were numerous in control rabbit optic nerve (mean 1879) and reduced in rabbits receiving the medium dose of pentoxifylline (300 mg PO BID, mean 439, p < 0.001) and the highest dose of pentoxifylline (600 mg PO BID, mean 120, p < 0.007). High dose pentoxifylline reduced TNF-alpha-induced axonal losses to less than 10% that seen without pentoxifylline pretreatment. Lower doses of pentoxifylline had a lesser but significant protective effect. Our results suggest that TNF-alpha-mediated axonal degeneration can be suppressed by high doses of pentoxifylline. Pentoxifylline may therefore be useful in AIDS patients demonstrating neurological or neuro ophthalmological symptoms. PMID- 9329036 TI - Basic fibroblast growth factor and platelet-derived growth factor prevent the death of spinal motor neurons after sciatic nerve transection in the neonatal rats. AB - In vivo, motor neurons are destined to die after axotomy. Several neuronal growth factors, such as ciliary neurotrophic factor, brain-derived neurotrophic factor, and leukemia inhibitory factor rescue neuronal death of axotomized motor neurons. Here, we report that systemically administered basic fibroblast growth factor and platelet-derived growth factor prevented spinal motor neuron death in neonatal rats following sciatic nerve resection. These data indicate that basic fibroblast growth factor and platelet derived growth factor play a role for motor neuron survival in vivo. PMID- 9329038 TI - Ebselen (DR3305) ameliorates delayed cerebral vasospasm in a canine two hemorrhage model. AB - Ebselen, a seleno-organic compound which inhibits arachidonic acid lipoxygenase activity and exerts glutathione peroxidase-like activity, ameliorated delayed cerebral vasospasm in a canine two-hemorrhage model. Twenty-five dogs were exposed to subarachnoid hemorrhage and divided into two groups. In the Ebselen treated group (6 dogs), 50 mg kg-1 of Ebselen was administered twice a day, for 7 days. The other 19 dogs without administration of Ebselen were used as a control group. In the Ebselen-treated group, the basilar artery on Day 7 after subarachnoid hemorrhage was constricted to 68.1 +/- 6.4% (mean +/- SD, n = 6) of the angiographic diameter on Day 0, before subarachnoid hemorrhage. This percentage was significantly larger than the 41.3 +/- 4.6% (n = 19) in the control group. The basilar artery segment obtained on Day 7 in the Ebselen treated group produced 3.32 +/- 1.82 nmol of 5-hydroxyeicosatetraenoic acid/mg protein/5 minutes (n = 5), significantly less than the 6.73 +/- 0.95 (n = 3) produced by the artery in the control group. Thus, administration of Ebselen suppressed arachidonate 5-lipoxygenase activation and had a beneficial effect on angiographically detected delayed vasospasm. PMID- 9329037 TI - Dysfunction of nitric oxide induces protein kinase C activation resulting in vasospasm after subarachnoid hemorrhage. AB - We hypothesize that the interaction between protein kinase C (PKC) and nitric oxide (NO) plays a role in the modulation of cerebral vascular tone, and the disturbance of this interaction following subarachnoid hemorrhage (SAH) results in vasospasm. To prove this hypothesis with direct evidence, PKC activities of smooth muscle cells of canine basilar arteries in the control and in the SAH groups were measured by an enzyme immunoassay method. N omega-nitro-L arginine (L NA), an inhibitor of NO production, enhanced PKC activity. This enhancement was inhibited neither by 8-bromo-guanosine 3',5'-cyclic monophosphate (8-bromo-cGMP) nor SIN-1, a NO releasing agent. PKC activity in the SAH was significantly higher than in the control; however, no further enhancement was produced with L-NA. In the SAH, PKC activity was not inhibited either by 8-bromo-cGMP or SIN-1. We conclude that NO maintains an appropriate vascular tone through inactivation of PKC, and that this effect is disturbed following SAH, resulting in PKC-dependent vascular contraction, such as vasospasm. On the other hand, once PKC has been activated, NO precursors do not inhibit PKC. These facts indicate NO inactivates PKC through the inhibition of phosphatidylinositol breakdown. PMID- 9329049 TI - Hibernation-induction trigger. I. Opioid-like effects of prairie dog plasma albumin on induced contractility of guinea pig ileum. AB - Studies have shown that plasma albumin fractions (PAFs) from hibernating mammals can inhibit induced contractility of the guinea pig ileum similarly to morphine. This study examined PAFs from two species of prairie dogs, one that undergoes natural seasonal hibernation (white-tailed, WT) and one that does not but can be induced to hibernate (black-tailed, BT). Dose-response curves of lyophilized PAF yielded IC50 values (mg) of 20.23 for summer WT, 15.53 for hibernating WT, 15.45 for summer BT, and 13.16 for winter-active BT. Winter samples from both species have IC50s lower than samples from summer animals, indicating greater potency of winter PAFs in suppressing guinea pig ileum contractility and therefore the presence of more opioid ligands in winter prairie dog plasma. Studies to elucidate receptor selectivity of PAF continue. PMID- 9329050 TI - Hibernation-induction trigger. II. In vitro effects of prairie dog plasma albumin on mouse vas deferens contractility. AB - Involvement of opioid molecules in hibernation is well established, with the delta opioid receptor implicated in hibernation induction. Previous studies have shown that plasma albumin fractions (PAFs) from hibernating mammals contain an uncharacterized ligand called "hibernation-induction trigger" (HIT), which causes inhibition of induced contractility in the guinea pig ileum (GPI). In part I of this study, we described effects of PAF from two species of prairie dogs on induced contractility of the GPI. In the present study (part II), we examine the response of the mouse vas deferens (MVD), which is populated with the delta receptor subtype, to increasing concentrations of PAF from the white-tailed prairie dog (WT) and the black-tailed prairie dog (BT). Dose-response curves of lyophilized PAF yielded IC50 values (mg) (mean dose that inhibits contractility to 50% of control) of 11.0 for summer WT, 10.6 for hibernating WT, 9.4 for summer BT, 12.2 for winter active BT, and 4.7 for winter hibernating BT. These results suggest that delta opioid (HIT) is present in both species throughout the calendar year and that the induction of hibernation may involve not only levels of opioid but also dynamic interactions between endogenous opioid and its receptors. PMID- 9329051 TI - Bilateral hypothalamic dopamine infusion in male Zucker rat suppresses feeding due to reduced meal size. AB - Lateral hypothalamic area dopamine activity (LHA-DA) appears to play a contributory role in regulating food intake, in particular, meal size. In this study we examined our hypothesis that bilateral LHA-DA injection induced depression of food intake via reduced meal size. Dopamine (11 mg/ml) or vehicle was infused into bilateral LHA at 0.5 microliter/h via two osmotic minipumps in six study or six control obese male Zucker rats for 13 days, respectively. Meal size, meal number, as well as food intake were continuously measured before, during, and after dopamine infusion. Intra-LHA-DA infusion significantly depressed food intake. The decreased food intake was solely caused by a significant and profound reduction in meal size. There was a modest compensatory rise in meal number that gradually increased food intake so that it reached control level on 10th dopamine infusion day. However, feeding pattern did not normalize until dopamine infusion ceased. The findings support our hypothesis that LHA-DA may participate in regulating meal size. Data also demonstrate that meal size and meal number are regulated in a reciprocal and independent manner to compensate for each other. PMID- 9329052 TI - Ketanserin and anxiety levels: influence of gender, estrous cycle, ovariectomy and ovarian hormones in female rats. AB - The influence of gender, estrous cycle, ovariectomy and ovarian hormones on the behavioral effects of the 5-HT2 receptor antagonist, ketanserin (KET), was studied. Intact males, female rats in the four stages of the estrous cycle and ovariectomized (OVX) female rats 14 days after surgery were used. The OVX rats received progesterone [PROG, 25 mg/kg, subcutaneously (SC)] and/or estradiol benzoate (EB, 10 micrograms/kg, SC). KET (3 mg/kg, SC) was injected 30 min before testing. All the animals were subjected to the following behavioral tests: exploration of an elevated plus-maze and retention of a passive-avoidance response. KET enhanced the exploration of the open arms in diestrous female rats but inhibited this behavior during the other stages of the cycle and in OVX rats injected either with oil or EB. This dose of KET was ineffective in males and in OVX rats injected with PROG. Furthermore, KET inhibited the retention of the passive avoidance response in males, in diestrous and metestrous female rats and in OVX rats injected with oil. In estrous females and in OVX rats injected with EB, KET enhanced the passive-avoidance response. These results demonstrate that the sensitivity to KET differs with the gender, estrous cycle and hormonal treatment and suggest that central serotonergic activity is influenced by the hormonal status of the animal. PMID- 9329053 TI - Behavioral alterations in male golden hamsters exposed to chlorodibromomethane. AB - A number of behavioral parameters in male golden hamsters (Mesocricetus auratus) exposed to chlorodibromomethane (CDBM) were registered over a 10-day (day -9 to day 0) pretest period and a 14-day (day 1 to day 14) test period beginning at 8 weeks of age. Whereas the subchronically treated group (5 mg/kg body weight) only showed significantly increased water bottle contacts on days 4-7 compared with vehicle controls, the acutely treated group (50 mg/kg body weight) exhibited significantly increased locomotor activity on days 3-6 and decreased wheel running at day 6 until day 9. In the open field, the acutely exposed males displayed significantly more flank-mark movements at days 4 and 7 than did the vehicle control animals. After day 9, the acutely treated males showed no differences in any parameter compared with the control males, indicating recovery. During the social confrontation on day 14, the subchronically exposed males bit and approached the intruder significantly less often than did the control males, indicating an impairment of selected social behavior affected by CDBM. Our results of low-level (5 mg/kg) behavioral effects contribute to the general characterization of CDBM and suggest that tests on social capabilities in behavioral toxicology provide significant results in a field of low-level and sublethal chronic application on small samples of experimental animals. PMID- 9329054 TI - Behavioral and neurochemical effects induced by subchronic l-deprenyl administration. AB - (-)Deprenyl was administered orally to rats for 15 days. In the staircase maze, a reduction of incorrect responses was observed at 0.9 mg/kg/day; higher or lower doses (3.5 or 0.35 mg/kg/day) were ineffective. In the same range of doses, the subchronic administration of (-)deprenyl did not modify the levels of norepinephrine, 5-hydroxytryptamine, 5-hydroxyindolacetic acid or the density and affinity of alpha-noradrenergic receptors in the cortex, olfactory system, hippocampus and striatum. An increase of the dopamine and a reduction of dihydroxyphenylacetic acid levels was observed only at the highest tested doses, at which no behavioral modification was observed. Only at 1.0 mg/kg/day did ( )deprenyl increase the acetylcholine (ACh) levels in the olfactory system, hippocampus and striatum. This neurochemical effect may be correlated to the behavioral effect observed in the same range of doses. We propose that this increase of ACh levels is determined by an activation of dopaminergic systems, resulting from the increase in the levels of PE caused by the inhibition of monoamine oxidase B (MAO-B) by (-)deprenyl. PMID- 9329055 TI - Reduction of dizocilpine and scopolamine-induced deficits in avoidance responding by SCH 54388, a metabolite of felbamate. AB - Felbamate (2-phenyl-1,3-propanediol dicarbamate) is a novel antiepileptic agent with a unique structure and mechanism of action, possibly involving binding sites at the N-methyl-D-aspartate receptor (NMDA) complex. A monocarbomate metabolite of felbamate (SCH 54388) was compared to felbamate using a mouse passive avoidance paradigm (PAR). SCH 54388 was markedly free of toxic side effects up to doses of 300 mg/kg, sc. SCH 54388 reduced the deficit-producing effects of either scopolamine, a cholinergic antagonist, or dizocilpine (MK-801), an NMDA receptor channel blocker, in a dose-dependent manner. The effective dose range of SCH 54388 was between 0.01 and 10 mg/kg, sc. SCH 54388 was also orally active at doses between 0.1 and 10 mg/kg. Felbamate also reduced scopolamine and dizocilpine antagonism, but was less potent than SCH 54388, reducing scopolamine induced deficits at 1 to 3 mg/kg, sc. in a dose-dependent manner and reducing deficits induced by dizocilpine at doses of 0.1 and 3 mg/kg, SC. The reduction of dizocilpine-induced deficits by felbamate was not dose dependent. These results suggest that SCH 54388 has a mechanism of action involving either directly or indirectly, glutaminergic and cholinergic central neuronal systems. PMID- 9329056 TI - Behavioral effects of trichloroethylene and tetrachloroethylene in mice. AB - This study was performed to clarify the toxicological profiles of trichloroethylene (TRCE) and tetrachloroethylene (TECE) when they are administered intraperitoneally in mice. The ED50 for loss of righting reflex were 2596 mg/kg in TRCE and 4209 mg/kg in TECE. TRCE and TECE impaired bridge test performance at 500 and 2000 mg/kg, respectively. An operant behavior performance was also inhibited by TRCE at 1000 mg/kg and by TECE at 2000 mg/kg. Both TRCE and TECE exhibited anticonflict effects in a Vogel-type task at 500 mg/kg. This effect was confirmed by the finding that TRCE exhibited anticonflict action in a Geller-type paradigm at 250 mg/kg and more, as did TRCE did at 1000 mg/kg. These results show that TRCE and TECE affect various behaviors in mice and suggest that conflict behaviors are one of the most sensitive behavioral indicators of the effects of these substances. The toxicological profiles of TRCE and TECE with respect to behavioral effects were very similar, and they can be classified in a single category. PMID- 9329057 TI - Decreased intake of a liquid diet in nonfood-deprived rats following intra-PVN injections of GLP-1 (7-36) amide. AB - I.c.v. administration of glucagon-like peptide-1 (7-36) amide (GLP-1) dose dependently suppresses food intake in rats, and induces activation of c-fos within rat paraventricular hypothalamus (PVN). The present study sought to determine whether GLP-1 (7-36) amide may act within the PVN by examining the effects of intra-PVN administration of GLP-1 (7-36) amide on food intake in rats. Adult male rats (n = 11) were prepared with indwelling guide cannulae aimed at the PVN. Rats were allowed access to a palatable liquid diet (Ensure) and water during a daily 60-min test period with intakes measured every 15 min. Intra-PVN administration of GLP-1 (7-36) amide (10, 50, 100 and 200 ng) did not alter latency to feed, but did suppress liquid diet intake over a 1-h testing period, as a function of dose. These results suggest that GLP-1 (7-36) amide may act, in part, to suppress feeding through interactions with cells within the PVN. PMID- 9329058 TI - A sucrose-based maintenance diet increases sensitivity to appetite suppressant effects of naloxone. AB - Rats maintained under restricted access to food (but at 100% free-feeding weights) received one of two diets in their home cages: a palatable sucrose-based diet, or regular chow (grain based diet), and could respond for either sucrose- or grain-based reinforcers under an FR 40 reinforcement schedule (crossover design). Naloxone (0, 0.1, 0.3, 1.0, and 3.0 mg/kg) was more potent in reducing operant-chamber responding in rats maintained on a sucrose-based diet in their home cages than those fed a grain-based diet, regardless of the type of pellets available in the operant chambers. Whereas naloxone decreased response rate over the session, it had no effect on initiation of responding. Results support the hypothesis that opioids are involved in the maintenance, but not the initiation of consummatory behavior. Furthermore, increased potency of naloxone following chronic ingestion of palatable food is similar to that observed following chronic opiate administration, suggesting a relationship between palatability and opioids. PMID- 9329059 TI - Effects of diet on sensitization to cocaine-induced stereotypy in female rats. AB - The progressive increase in cocaine-induced stereotyped behavior that accompanies repeated cocaine injections (sensitization) was examined in rats consuming different diets. Adult female Sprague-Dawley rats were fed one of three diets: low protein (6% casein), adequate protein (25% casein), or a standard chow diet. Following 1 week of adaptation to the diets, the rats were injected every 3-4 days with either cocaine (30 mg/kg, IP) or saline, and the total amount of stereotypy was measured over a 90-min interval following each of four injections. Cocaine-induced stereotypy peaked at 40-50 min following each injection, after which it declined for all diet groups. With repeated injections, the total amount of stereotypy increased in all diet groups. By the fourth injection, the low protein diet group (6% casein) exhibited a slower onset and a possibly prolonged duration of cocaine-induced stereotypy when compared with the two adequate protein diet groups (25% casein and chow). Interestingly, the rats in the two purified diet groups (6% casein and 25% casein) exhibited significantly more stereotypy across injections than those in the chow diet group. Weight differences did not explain the differences in stereotypy present among the diet groups. This study concludes that diet significantly alters the pattern of cocaine-induced stereotypy in female rats, especially after repeated exposure. PMID- 9329060 TI - Cysteamine blocks amphetamine-induced deficits in sensorimotor gating. AB - Somatostatin is a neuropeptide that has been shown to interact with dopamine. Low concentrations of cysteamine selectively depletes somatostatin and has been used to investigate the role of endogenous somatostatin in lieu of an available selective receptor antagonist. We examined the effects of various doses of subcutaneous cysteamine on baseline and amphetamine-disrupted sensorimotor gating as measured by prepulse inhibition of the acoustic startle reflex. Cysteamine in doses ranging from 50-300 mg/kg reversed decreases in PPI induced by systemic injections of amphetamine (2 mg/kg). Cysteamine had no effect on the amplitude of the acoustic startle reflex itself. The results lend further support to a somatostatin-dopamine interaction within the brain in which endogenous somatostatin facilitates dopaminergic activity. These findings also suggest that endogenous somatostatin might play a significant role in regulation of sensorimotor gating deficits. This has clinical implications as deficient prepulse inhibition is recorded in humans suffering from neuropsychiatric conditions such as schizophrenia. PMID- 9329061 TI - Nicotine abstinence syndrome precipitated by central but not peripheral hexamethonium. AB - A rodent model of nicotine dependence has been developed based on continuous subcutaneous (s.c.) infusion of nicotine tartrate. Nicotine abstinence syndrome was precipitated by s.c. injection of the nicotinic antagonist mecamylamine, which freely crosses the blood-brain barrier. In contrast, the nicotinic antagonist hexamethonium crosses the blood-brain barrier very poorly. This study determined whether central or peripheral administration of hexamethonium could precipitate nicotine abstinence. In the first experiment, 26 nicotine-dependent rats were injected s.c. with 0.5, 5 or 10 mg/kg hexamethonium dichloride or saline alone and observed for 20 min. Few abstinence signs were observed in any group; there was no significant drug effect. In the second experiment, 18 rats were cannulated in the third ventricle and rendered nicotine dependent. One week later, rats were injected through the cannula with 12 or 18 ng hexamethonium or saline alone and observed for 20 min. Both dose groups differed significantly from the saline-injected group, and there was a significant positive linear trend of signs as a function of dose. The high dose had no significant effect in 14 nondependent rats. We conclude that hexamethonium is much more potent by the central route, and there is a major central nervous system component in nicotine dependence. PMID- 9329062 TI - Effects of salicylate on 3,4-methylenedioxymethamphetamine (MDMA)-induced neurotoxicity in rats. AB - The drug 3,4-methylenedioxymethamphetamine (MDMA) is a serotonergic neurotoxicant that causes hyperthermia and depletion of serotonin (5-HT) and 5-hydroxy-indole-3 acetic acid (5-HIAA) in the central nervous system. Formation of neurotoxic metabolites of MDMA, e.g., 2,4,5-trihydroxy-methamphetamine and 2,4,5 trihydroxyamphetamine, involves hydroxyl and/or superoxide free radicals. The present study was designed to determine whether the hydroxyl free-radical trapping agent salicylate could provide protection against MDMA neurotoxicity in rats. In the acute studies, sodium salicylate (12.5-400 mg/kg, calculated as free acid) was injected interperitoneally (i.p.) 1 h before subcutaneous (s.c.) injections of MDMA (20 mg/kg as base). In the chronic studies, sodium salicylate (3.1-100 mg/kg) was injected i.p. 1 h before repeated s.c. injections of MDMA (10 mg/kg as base, twice daily, at 0830 and 1730 h for 4 consecutive days). Repeated MDMA administration depleted contents of 5-HT and 5-HIAA in the frontal cortex, hippocampus and striatum. Coadministration of salicylate plus MDMA did not significantly alter MDMA-induced depletion of 5-HT and 5-HIAA in these tissues. Thus, salicylate, a hydroxyl free-radical-trapping agent, does not protect against MDMA-induced hyperthermia and depletion of 5-HT and 5-HIAA. These observations suggest that MDMA-induced neurotoxicity may occur mainly through the production of superoxide or other radicals rather than hydroxyl free radicals. Salicylate actually potentiated MDMA-induced hyperthermia and lethality, findings that might be of clinical relevance. PMID- 9329063 TI - The antiseizure efficacies of MK-801, phencyclidine, ketamine, and memantine are altered selectively by stress. AB - Adaptive changes in the NMDA receptor complex occur in response to exposure to stress. We have previously shown that the ability of MK-801, an uncompetitive NMDA receptor antagonist, to antagonize electrically precipitated tonic hind-limb extension is reduced 24 h after mice are forced to swim for up to 10 min in cold water. The stress-induced reduction of the antiseizure efficacy of MK-801 stimulated the proposal that mice exposed to swim stress may serve as "an intact animal model" of altered or diminished NMDA-mediated neural transmission. In the current investigation, the dose-dependent abilities for the antagonism of electrically precipitated seizures in mice were determined for MK-801, phencyclidine, ketamine, and memantine. Interestingly, a single session of cold water swim stress reduced the antiseizure efficacies of MK-801 and memantine without affecting phencyclidine and ketamine when tested 24 h later. The data do not suggest that stress results in a simple reduction in the number of activated or open channels, but rather alters their size or charge characteristics. PMID- 9329064 TI - Effects of neonatal exposure to nicotine on electrophysiological parameters in adult rats. AB - In clinical studies and animal models, there is evidence that nicotine exposure during gestation can result in deficits in cognitive performance. The present study examined the effects of two doses of neonatal nicotine exposure on adult brain activity as assessed by the N1 and P3 components of the event-related potential (ERP) and background electroencephalography (EEG). Nicotine (0 mg, 1 mg/kg/day, 4 mg/kg/day) was administered to neonatal rat pups from postnatal day 4 (PN4) through PN12 with an artificial rearing paradigm; suckled rats served as additional control subjects. Nicotine exposure was specifically found to alter responses of the P3 component of the ERP, recorded in dorsal hippocampus, to changes in stimulus parameters. A significant reduction in the response of the P3A component to the noise tone as compared with the level of the frequently presented tone was found. A significant reduction in the response to the noise tone as compared with the level of the infrequently presented tone also was seen in the P3B component. No effects of drug exposure were found on the N1 component in any lead, although artificial rearing produced specific effects on the latency of the N1 component in cortex. No significant differences among treatment groups were found on any of the EEG-dependent variables. Female rats overall were found to have significantly higher EEG amplitudes than the males, a finding previously reported in our laboratory. However, no overall effects of gender were found on any ERP component. These studies suggest that neonatal nicotine exposure specifically reduces the electrophysiological response of the hippocampus to changes in auditory stimuli. Additional studies will be necessary to link these P3 amplitude changes to the effects of nicotine on the developing brain in human and animal subjects. PMID- 9329065 TI - Absorption and subjective effects of caffeine from coffee, cola and capsules. AB - Coffee is often perceived as producing greater pharmacological effects than cola. The present study compared the magnitude and rapidity of peak caffeine levels and subjective effects between coffee and cola. Thirteen users of both coffee and cola (mean daily caffeine consumption = 456 mg) ingested 400 mg caffeine via 12 oz unsweetened coffee, 24 oz sugar-free cola or 2 capsules in a random, double blind, placebo-controlled, within-subjects design. Subjects provided a saliva sample and completed subjective effect scales 15 min before and 30, 60, 90, 120, 180 and 240 min after ingestion. Mean peak saliva caffeine levels did not differ between coffee (9.7 +/- 1.2 micrograms/ml) and cola (9.8 +/- 0.9 micrograms/ml) and appeared to be greater with these beverages than with the capsule (7.8 +/- 0.6 micrograms/ml; p = NS). Saliva caffeine levels peaked at similar times for coffee (42 +/- 5 min) and cola (39 +/- 5 min) but later for capsule (67 +/- 7 min; p = 0.004). There was no main effect of vehicle or interaction of vehicle and drug on magnitude of peak effect or time to peak increase on self-report scales. In summary, peak caffeine absorption, time to peak absorption, and subjective effects do not appear to be influenced by cola vs. coffee vehicle. Perceived differences in the effects of coffee vs. cola may be due to differences in dose, time of day, added sweetener, environmental setting or contingencies. PMID- 9329066 TI - Drug disruption of short-term memory in Drosophila melanogaster. AB - Recent work on operant visual learning and memory in Drosophila has suggested at least three distinct memory phases. Trying to disrupt memory pharmacologically, we fed flies with ouabain or the depolarizing drugs potassium chloride (KCl), lithium chloride (LiCl) and monosodium glutamate for some specific time before training. The depolarizing drugs abolished memory very soon after training. Ouabain exerted no effect on memory within the first 20 min but abolished it more than 30 min after training. These drugs had no diminishing effects on the visual discrimination and behavioral performance of the flies during training. This result suggests that memory disruption may not be induced by nonspecific effects of the drugs. In addition, reversal training of the KCl-fed flies indicates that KCl appears not to impair the retrieval mechanism of flies. These results suggest that the specific disruptive effects of the drugs on memory formation and the existence of a short-term memory phase, are susceptible to disruption of the depolarizing drugs but unaffected by ouabain. PMID- 9329067 TI - Anxiogenic behavior in the light-dark paradigm follwoing intraventricular administration of cholecystokinin-8S, restraint stress, or uncontrollable footshock in the CD-1 mouse. AB - The influence of restraint stress (0, 15, 30, or 60 min), uncontrollable footshock (0, 15, 30, or 60 shocks), or intraventricular CCK-8S administration (0, 5, 25, or 50 ng delivered in a 1 microliter volume) were evaluated on transition frequency and cumulative time in light among CD-1 mice in the light dark paradigm. Mice exposed to restraint stress of either 15 or 60 min were indistinguishable from nonrestrained animals, while the 30-min session of restraint decreased time in light and transition scores. The presentation of 15, 30, or 60 uncontrollable footshocks were equally effective in decreasing cumulative time in light but had no effect on transition scores. Intraventricular infusion of 25 and 50 ng doses of cholecystokinin-8S reduced cumulative time in light and transition frequency in CD-1 mice relative to vehicle or 5 ng CCK-8S treated animals in the light-dark paradigm. The time in light and transition data secured among mice with repeated light-dark exposure and 30 min of restraint were comparable to the corresponding scores secured when performance was only evaluated on trial 1. Transition scores were reduced on trial 1 of mice exposed to 30 min of footshock, but time in light was reminiscent of the performance detected among mice with prior light-dark experience. Potential neurochemical correlates associated with the anxiogenic effects associated with stressor exposure and CCK-8S administration in the light-dark task are discussed. PMID- 9329068 TI - Anxiolytic action of a neurokinin1 receptor antagonist in the social interaction test. AB - CGP 49823, a substance P antagonist acting at NK1 receptors, had significant anxiolytic effects at 3, 10 and 30 mg/kg orally in the high-light unfamiliar and low-light unfamiliar conditions of the social interaction test but was without effect in the low-light familiar condition. The effects were less marked after 3 and 6 weeks of treatment (10 mg/kg/day), indicating that some tolerance had developed, but a significant anxiolytic effect still remained. After 3 weeks of diazepam treatment (2 mg/kg/day, intraperitoneally), tolerance developed to the anxiolytic effects, and there was an anxiogenic response 24 h after withdrawal. In contrast, there were no anxiogenic withdrawal effects 24 h after 3 weeks or 24, 48 or 72 h after 6 weeks treatment with CGP 49823 (10 mg/kg/day). These results suggest that the compound may prove to be a useful anxiolytic and that substance P may play a role in mediating states of anxiety. PMID- 9329069 TI - L-NNA decreases cortical vascularization, alcohol preference and withdrawal in alcoholic rats. AB - Rats, which were made chronically alcoholic in combination with L-N(o)-nitro arginine (L-NNA) treatment (5 mg/kg/day), a nitric oxide (NO) synthase inhibitor, showed a significant decrease in their alcohol preference and hypermotility during the withdrawal period by comparison with chronically alcoholic rats. However, no difference in the global liquid consumption between treated and untreated rats during the withdrawal stage was identified. In addition, the hypervascularization of the cortical area observed after chronic alcoholism was significantly decreased in the rats that had received L-NNA during the alcoholism procedure and was comparable to control rats. Thus, L-NNA alters both the behavioral preference for alcohol after alcoholism and the hypermotility during alcohol withdrawal, thus supporting the hypothesis of a direct implication of NO in alcohol abuse and its withdrawal. PMID- 9329070 TI - Differences between two substrains of AB mice in the opioid system. AB - Animals from two substrains of AB mice, i.e., ABH/Md and ABG/Md, differ in the occurrence of aggressive behavior. After maturation, male ABH mice regularly exhibited abnormal aggressive behavior making group-housing impossible. In contrast, ABG animals never showed such behavioral patterns. To elucidate the role of opioid mechanisms, we tested the reaction of these animals to morphine in the hot plate test. Moreover, specific DAMGO binding was measured. It was shown that mice from control groups differed significantly in reaction to the thermal stimulus. ABH mice had significantly longer reaction times. With increasing doses of morphine this difference disappeared, suggesting different levels of basal activity in endogenous opioid systems. This is underlined by significantly lower DAMGO binding in aggressive ABH mice. The results suggest that differences in endogenous opioid systems may account for differences in aggressiveness. PMID- 9329071 TI - Fluoxetine decreases fat and protein intakes but not carbohydrate intake in male rats. AB - Administration of fluoxetine, a selective serotonin reuptake inhibitor, results in decreases in food intake and body weight. The present study investigated whether the anorectic actions of fluoxetine were due to a general decrease in caloric intake or macronutrient specific. Male Long-Evans rats were maintained on a dietary self-selection regime with separate sources of protein, fat, and carbohydrate. During the acute phase of the experiment, nutrient intakes were measured 2, 4, 6, and 24 h after injections of 0, 5.0, and 10.0 mg/kg fluoxetine hydrochloride. Fluoxetine significantly decreased protein and fat intakes in a dose-related manner at all measurement times. In comparison, fluoxetine had a less pronounced effect on carbohydrate intake. During the chronic phase, rats were divided into two groups, one receiving daily injections of 10.0 mg/kg fluoxetine, and the other, vehicle injections. Drug injections continued for 28 days, and were followed by a 28-day withdrawal period. Rats given fluoxetine on a chronic basis consumed significantly less calories and gained significantly less weight than rats injected with the vehicle. Both caloric intake and body weight returned to control values during the withdrawal period. Fat and protein intakes also were significantly reduced throughout the drug injection period, and were restored to baseline levels during the withdrawal period. In contrast, carbohydrate intake was not reduced on an absolute basis, and actually was increased as percent of total caloric intake during the drug period. The results of this experiment call into question the idea that increased serotoninergic activity is related to selective reductions in carbohydrate intake. PMID- 9329073 TI - Serum estradiol levels and ethanol-induced aggression. AB - The biological mechanisms behind ethanol-induced aggression are not known. Because gonadal hormones are linked both to aggression and ethanol, the present study examined relationships among the levels of serum estradiol (E2), testosterone (T), and aggressive behavior in ethanol-treated male mice. We found that among group-housed male mice, serum E2 levels were significantly elevated 30 min after a single injection of 0.6 g/kg ethanol. Serum T levels showed a nonsignificant decrease by ethanol. The E2/T ratio, an index of aromatization of T to E2, was significantly higher in the ethanol-treated animals when compared with the vehicle-treated animals. We also determined aggressive behavior in the resident-intruder test among isolated male mice at baseline (after a vehicle), and after an injection of 0.6 g/kg ethanol. The mice were grouped accordingly to those that increased, decreased, or remained nonaggressive in response to ethanol administration. We found that at baseline, neither serum T or E2 levels, nor E2/T ratio differed significantly between the increased or reduced aggressor mice. In contrast to the increase in serum E2 levels seen in the nonaggressive mice, ethanol significantly reduced circulating E2 levels, but did not affect aromatization of E2 from T in the mice that became aggressive following an ethanol injection. These data suggest that mice who exhibit a paradoxical decrease in serum E2 levels by ethanol may be particularly prone to ethanol induced aggression. PMID- 9329072 TI - Effect of aversive stimulation on 5-hydroxytryptamine and dopamine metabolism in the rat brain. AB - The neurochemical consequences of aversive behavior based on novelty, rat social interaction, have been assessed in various rat brain regions utilizing high performance liquid chromatography coupled with an electrochemical detector (HPLC ECD) technique. The present studies indicated that compared to animals from the home cage, those exposed to the high-light aversive unfamiliar test condition, had significantly increased levels of 5-hydroxyindoleacetic acid (5-HIAA), the metabolite of 5-hydroxytryptamine (5-HT), in the tested brain regions including amygdala, entorhinal cortex, frontal cortex, temporal cortex, tuberculum olfactorium, hippocampus, nucleus accumbens, and striatum. The levels of 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), the metabolites of dopamine (DA), were increased in tuberculum olfactorium, nucleus accumbens, and striatum. When compared to the low-light familiar test condition (LF), the levels, following exposure to the highlight unfamiliar situation, of 5-HIAA were significantly increased in the amygdala, entorhinal cortex, tuberculum olfactorium, hippocampus, and nucleus accumbens, while the 5-HIAA levels remained unchanged in the frontal cortex, temporal cortex, and striatum. The DOPAC and HVA levels were also increased by the HU situation in the amygdala, tuberculum olfactorium, and nucleus accumbens. An increase was also found for the levels of DA in the amygdala. Such effects were prevented by diazepam or the 5-HT3 receptor antagonist ondansetron. It is concluded that the aversive test condition of the social interaction test (HU) increases 5-HT and DA turnover throughout the rat brain. Such effects might be related to the sensitivity to novel anxiolytic drug of the social interaction test. PMID- 9329074 TI - Chronic cocaine exposure affects stimulus-induced but not spontaneous behavior of the near-term mouse fetus. AB - Pregnant female mice were injected subcutaneously with a 40-mg/kg dose of cocaine HCl or physiological saline from day 1 through day 17 of gestation. On day 18 of gestation, dams were surgically prepared to allow the behavior of their fetuses to be observed. Spontaneous motor behavior was unaffected by cocaine exposure. Cocaine exposure potentiated motor responses of the fetuses to ammonia and to control injections of saline into the amniotic sac. Restriction of umbilical blood flow caused a specific stereotyped response in saline-injected fetuses, which is in agreement with studies of other species. This response was markedly potentiated in fetuses exposed to cocaine. The results suggest that the mouse may be a viable model for studies of the neurodevelopmental effects of gestational cocaine exposure and are discussed in relation to current models of the effects of long-term cocaine exposure on brain neurochemistry. PMID- 9329075 TI - Pimozide does not shift palatability: separation of anhedonia from sensorimotor suppression by taste reactivity. AB - Several "taste reactivity" studies of dopamine and reward have concluded that pimozide suppresses the hedonic reaction patterns normally elicited by sucrose but enhances aversive reaction patterns elicited by quinine. However, other taste reactivity studies have failed to find hedonic/aversive shifts in reaction patterns after dopamine antagonists or dopamine lesions. The divergent conclusions have come from two different laboratories. To resolve the controversy regarding dopamine blockade and palatability, the present study joined the two laboratories to investigate the effect of pimozide on taste reactivity patterns elicited by sucrose and quinine. The results replicated many (but not all) of the earlier findings and identified procedural factors responsible for different outcomes. Overall, the results provide evidence for sensorimotor effects of pimozide on taste reactivity but not for a hedonic shift in palatability. Pimozide suppressed both hedonic and aversive reaction patterns in a gradual sensorimotor fashion when the eliciting taste stimulus was repeated or continued for several minutes. The general suppression typically did not alter the initial reaction to a taste but emerged only after an oral infusion of sucrose or quinine continued for several minutes or trials. Aversive reactions were never enhanced. The balance between hedonic and aversive reaction patterns was not shifted by pimozide. We conclude that pimozide produces a sensorimotor impairment of taste reactivity patterns but does not shift taste palatability toward anhedonia or aversion. PMID- 9329076 TI - Pharmacological characterization of the discriminative stimulus effects of clenbuterol in rats. AB - The beta-2 selective adrenergic agonist clenbuterol produces discriminative stimulus effects in rats. Administration of beta adrenergic agonists that do not cross the blood-brain barrier well following peripheral administration either failed to substitute for clenbuterol or resulted in chance levels of drug appropriate responding; this suggested central mediation of the effects of clenbuterol. This interpretation was supported by the finding that the centrally acting beta adrenergic antagonist propranolol antagonized the discriminative stimulus effects of clenbuterol more potently than did CGP-12177, a hydrophilic beta adrenergic antagonist that has been shown to have very limited central activity. Antagonism experiments using subtype-selective antagonists showed that the beta-2 selective antagonist ICI 118,551 more potently antagonized the discriminative effects of the training dose of clenbuterol than did the beta-1 selective antagonist betaxolol. The present results indicate that the discriminative stimulus effects of clenbuterol provide an in vivo index of activation of central beta-2 adrenergic receptors. PMID- 9329077 TI - Dopamine infusion does not alter LH levels before or after chronic cocaine exposure in female rhesus monkeys. AB - Cocaine stimulates release of luteinizing hormone (LH) in preclinical and clinical studies but the contribution of the indirect dopamine agonist actions of cocaine to its effects on LH are unclear. In the present study, we examined the effects of exogenous dopamine infusions on LH release in drug-naive, normally cycling, female rhesus monkeys. All studies were conducted during the mid follicular phase (cycle days 6-8). Three successive 80-min dopamine infusions (10 micrograms/kg/min, intravenous) were alternated with 20- or 40-min interruptions of dopamine infusions. There were no significant changes in LH during or following dopamine infusions. Predopamine baseline LH levels averaged 30 +/- 5.4 ng/ml. LH averaged 31.7 +/- 1.3 ng/ml during dopamine infusions and 31.4 +/- 1.3 ng/ml after dopamine infusions stopped. To determine whether chronic cocaine exposure influenced the effect of dopamine on LH, rhesus females were studied after more than 2 years of cocaine self-administration at an average dose of 6.5 +/- 0.2 mg/kg/day. LH averaged 27.3 +/- 3.3 ng/ml during baseline and 26.9 +/- 0.7 ng/ml and 26.1 +/- 0.7 ng/ml during dopamine infusions and interruptions, respectively. Similarly, during withdrawal from cocaine, baseline LH levels averaged 32.1 +/- 4.5 ng/ml, and LH did not change significantly during dopamine infusions (31.2 +/- 1.1 ng/ml) and infusion interruptions (32.1 +/- 1.1 ng/ml). Under the conditions of the present study, dopamine administration did not change LH levels in gonadally intact rhesus monkeys, and these findings are consistent with previous studies in ovariectomized rhesus females. However, these data are not consistent with clinical reports, and some possible implications of this species difference are discussed. Moreover, these data suggest that the stimulation of LH by cocaine may not be explained by its indirect dopamine agonist actions. PMID- 9329078 TI - Backbone entropy of loops as a measure of their flexibility: application to a ras protein simulated by molecular dynamics. AB - The flexibility of surface loops plays an important role in protein-protein and protein-peptide recognition; it is commonly studied by Molecular Dynamics or Monte Carlo stimulations. We propose to measure the relative backbone flexibility of loops by the difference in their backbone conformational entropies, which are calculated here with the local states (LS) method of Meirovitch. Thus, one can compare the entropies of loops of the same protein or, under certain simulation conditions, of different proteins. These loops should be equal in size but can differ in their sequence of amino acids residues. This methodology is applied successfully to three segments of 10 residues of a Ras protein simulated by the stochastic boundary molecular dynamics procedure. For the first time estimates of backbone entropy differences are obtained, and their correlation with B factors is pointed out; for example, the segments which consist of residues 60-65 and 112 117 have average B factors of 67 and 18 A2, respectively, and entropy difference T delta S = 5.4 +/- 0.1 kcal/mol at T = 300 K. In a large number of recent publications the entropy due to the fast motions (on the ps-ns time scale) of N-H and C-H vectors has been obtained from their order parameter, measured in nuclear magnetic resonance spin relaxation experiments. This enables one to estimate differences in the entropy of protein segments due to folding-unfolding transitions, for example. However, the vectors are assumed to be independent, and the effect of the neglected correlations is unknown; our method is expected to become an important tool for assessing this approximation. The present calculations, obtained with the LS method, suggest that the errors involved in experimental entropy differences might not be large; however, this should be verified in each case. Potential applications of entropy calculations to rational drug design are discussed. PMID- 9329079 TI - Crystal structures of the methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath): implications for substrate gating and component interactions. AB - The crystal structure of the nonheme iron-containing hydroxylase component of methane monooxygenase hydroxylase (MMOH) from Methylococcus capsulatus (Bath) has been solved in two crystal forms, one of which was refined to 1.7 A resolution. The enzyme is composed of two copies each of three subunits (alpha 2 beta 2 gamma 2), and all three subunits are almost completely alpha-helical, with the exception of two beta hairpin structures in the alpha subunit. The active site of each alpha subunit contains one dinuclear iron center, housed in a four-helix bundle. The two iron atoms are octahedrally coordinated by 2 histidine and 4 glutamic acid residues as well as by a bridging hydroxide ion, a terminal water molecule, and at 4 degrees C, a bridging acetate ion, which is replaced at -160 degrees C with a bridging water molecule. Comparison of the results for two crystal forms demonstrates overall conservation and relative orientation of the domain structures. The most prominent structural differences identified between the two crystal forms is in an altered side chain conformation for Leu 110 at the active site cavity. We suggest that this residue serves as one component of a hydrophobic gate controlling access of substrates to and products from the active site. The leucine gate may be responsible for the effect of the B protein component on the reactivity of the reduced hydroxylase with dioxygen. A potential reductase binding site has been assigned based on an analysis of crystal packing in the two forms and corroborated by inhibition studies with a synthetic peptide corresponding to the proposed docking position. PMID- 9329080 TI - Variations on a theme by Debye and Waller: from simple crystals to proteins. AB - Debye and Waller showed how to adjust scattering intensities in diffraction experiments for harmonic motions of atoms about an average, static reference configuration. However, many motions, particularly in biological molecules as compared to simple crystals, are far from harmonic. We show how, using a variety of simple anharmonic, multiconformational models, it is possible to construct a variety of Generalized Debye-Waller Factors, and understand their meaning. A central result for these cases is that, in principle, intensity factors cannot be obtained from true total mean square displacements of the atoms. We make the distinction between the intensity factors for unimodal quasiharmonic motions and those for the anharmonic, multimodal (valley hopping) motions. Only the former affect the conventional B factors. PMID- 9329081 TI - VL:VH domain rotations in engineered antibodies: crystal structures of the Fab fragments from two murine antitumor antibodies and their engineered human constructs. AB - The crystal structures of two pairs of Fab fragments have been determined. The pairs comprise both a murine and an engineered human form, each derived from the antitumor antibodies A5B7 and CTM01. Although antigen specificity is maintained within the pairs, antigen affinity varies. A comparison of the hypervariable loops for each pair of antibodies shows their structure has been well maintained in grafting, supporting the canonical loop model. Detailed structural analysis of the binding sites and domain arrangements for these antibodies suggests the differences in antigen affinity observed are likely to be due to inherent flexibility of the hypervariable loops and movements at the VL:VH domain interface. The four structures provide the first opportunity to study in detail the effects of protein engineering on specific antibodies. PMID- 9329082 TI - Understanding the recognition of protein structural classes by amino acid composition. AB - Knowledge of amino acid composition, alone, is verified here to be sufficient for recognizing the structural class, alpha, beta, alpha + beta, or alpha/beta of a given protein with an accuracy of 81%. This is supported by results from exhaustive enumerations of all conformations for all sequences of simple, compact lattice models consisting of two types (hydrophobic and polar) of residues. Different compositions exhibit strong affinities for certain folds. Within the limits of validity of the lattice models, two factors appear to determine the choice of particular folds: 1) the coordination numbers of individual sites and 2) the size and geometry of non-bonded clusters. These two properties, collectively termed the distribution of non-bonded contacts, are quantitatively assessed by an eigenvalue analysis of the so-called Kirchhoff or adjacency matrices obtained by considering the non-bonded interactions on a lattice. The analysis permits the identification of conformations that possess the same distribution of non-bonded contacts. Furthermore, some distributions of non bonded contacts are favored entropically, due to their high degeneracies. Thus, a competition between enthalpic and entropic effects is effective in determining the choice of a distribution for a given composition. Based on these findings, an analysis of non-bonded contacts in protein structures was made. The analysis shows that proteins belonging to the four distinct folding classes exhibit significant differences in their distributions of non-bonded contacts, which more directly explains the success in predicting structural class from amino acid composition. PMID- 9329083 TI - Studies on the inhibitor-binding site of porcine aldehyde reductase: crystal structure of the holoenzyme-inhibitor ternary complex. AB - Aldehyde reductase is an enzyme capable of metabolizing a wide variety of aldehydes to their corresponding alcohols. The tertiary structures of aldehyde reductase and aldose reductase are similar and consist of an alpha/beta-barrel with the active site located at the carboxy terminus of the strands of the barrel. We have determined the X-ray crystal structure of porcine aldehyde reductase holoenzyme in complex with an aldose reductase inhibitor, tolrestat, at 2.4 A resolution to obtain a picture of the binding conformation of inhibitors to aldehyde reductase. Tolrestat binds in the active site pocket of aldehyde reductase and interacts through van der Waals contacts with Arg 312 and Asp 313. The carboxylate group of tolrestat is within hydrogen bonding distance with His 113 and Trp 114. Mutation of Arg 312 to alanine in porcine aldehyde reductase alters the potency of inhibition of the enzyme by aldose reductase inhibitors. Our results indicate that the structure of the inhibitor-binding site of aldehyde reductase differs from that of aldose reductase due to the participation of nonconserved residues in its formation. A major difference is the participation of Arg 312 and Asp 313 in lining the inhibitor-binding site in aldehyde reductase but not in aldose reductase. PMID- 9329084 TI - A molecular dynamics simulation study of segment B1 of protein G. AB - The immunoglobulin binding protein, segment B1 of protein G, has been studied experimentally as a paradigm for protein folding. This protein consists of 56 residues, includes both beta sheet and alpha helix and contains neither disulfide bonds nor proline residues. We report an all-atom molecular dynamics study of the native manifold of the protein in explicit solvent. A 2-ns simulation starting from the nuclear magnetic resonance (NMR) structure and a 1-ns control simulation starting from the x-ray structure were performed. The difference between average structures calculated over the equilibrium portion of trajectories is smaller than the difference between their starting conformations. These simulation averages are structurally similar to the x-ray structure and differ in systematic ways from the NMR-determined structure. Partitioning of the fluctuations into fast (< 20 ps) and slow (> 20 ps) components indicates that the beta sheet displays greater long-time mobility than does the alpha helix. Clore and Gronenborn [J. Mol. Biol. 223:853-856, 1992] detected two long-residence water molecules by NMR in a solution structure of segment B1 of protein G. Both molecules were found in the fully exposed regions and were proposed to be stabilized by bifurcated hydrogen bonds to the protein backbone. One of these long-residence water molecules, found near an exposed loop region, is identified in both of our simulations, and is seen to be involved in the formation of a stable water-mediated hydrogen bond bridge. The second water molecule, located near the middle of the alpha helix, is not seen with an exceptional residence time in either as a result of the conformation being closer to the x-ray structure in this region of the protein. PMID- 9329085 TI - Structure of the LAV6 peptide: a nucleation site for the correct receptor-induced refolding of the CD4-binding domain of HIV1 gp 120. AB - LAV44 and LAV15 (lymphadenopathy-associated virus) peptides of the CD4-binding region of gp 120 per se bind to the CD4 receptor (Reed and Kinzel, Biochemistry 30: 4521-4528, 1991; Lasky et al., Cell 50:975-985, 1987). Depending on the environment, the LAV peptides exhibit the ability to switch cooperatively between beta-sheet and helical conformation when solvent polarity is changed past a critical point. This property, which is dependent on the amino acid sequence LPCR, is crucial for receptor binding (Reed and Kinzel, Proc. Natl. Acad. Sci. U.S.A. 90:6761-6765, 1993). Structure determination with 2D-NMR-spectroscopy reveals that LAV6 peptide (sequence: TLPCRI) has a well-defined structure, partially exhibiting inverse gamma-turn conformation in aqueous solution. Quantitative evaluation of the NMR data discloses 90% trans-conformation for the peptide bond between leucine and proline. The psi- and phi-angles fall into the typical range for amino acids located in turns. On the other hand, the amino acid sequence C-terminal to the LPCR tetrad has been shown to fold atypically in the absence of these residues. All these results show that the short sequence of LAV6 peptide, with the central amino acids LPCR, displays a matrix-independent structure and may, therefore, act as a conformational template for forming secondary structure in the intact CD4-binding domain of gp 120. PMID- 9329086 TI - Refolding simulations of an isolated fragment of barnase into a native-like beta hairpin: evidence for compactness and hydrogen bonding as concurrent stabilizing factors. AB - Experimental evidence and theoretical models both suggest that protein folding is initiated within specific fragments intermittently adopting conformations close to that found in the protein native structure. These folding initiation sites encompassing short portions of the protein are ideally suited for study in isolation by computational methods aimed at peering into the very early events of folding. We have used Molecular Dynamics (MD) technique to investigate the behavior of an isolated protein fragment formed by residues 85 to 102 of barnase that folds into a beta hairpin in the protein native structure. Three independent MD simulations of 1.3 to 1.8 ns starting from unfolded conformations of the peptide portrayed with an all-atom model in water were carried out at gradually decreasing temperature. A detailed analysis of the conformational preferences adopted by this peptide in the course of the simulations is presented. Two of the unfolded peptides conformations fold into a hairpin characterized by native and a larger bulk of nonnative interactions. Both refolding simulations substantiate the close relationship between interstrand compactness and hydrogen bonding network involving backbone atoms. Persistent compactness witnessed by side-chain interactions always occurs concomitantly with the formation of backbone hydrogen bonds. No highly populated conformations generated in a third simulation starting from the remotest unfolded conformer relative to the native structure are observed. However, nonnative long-range and medium-range contacts with the aromatic moiety of Trp94 are spotted, which are in fair agreement with a former nuclear magnetic resonance study of a denaturing solution of an isolated barnase fragment encompassing the beta hairpin. All this lends reason to believe that the 85-102 barnase fragment is a strong initiation site for folding. PMID- 9329087 TI - Correlation of the enzyme activities of Bacillus stearothermophilus lactate dehydrogenase on three substrates with the results of molecular dynamics/energy minimization conformational searching. AB - Current methods for reengineering enzyme substrate specificities rely heavily on the use of static x-ray crystallographic models. In this article we detail the use of a molecular mechanics approach for suggesting regions of Bacillus stearothermophilus L-lactate dehydrogenase (EC 1.1.1.27) involved in substrate specificity, and hence areas of interest for protein engineers. The approach combines molecular dynamics with energy minimization (MD/EM) to search the conformational space available to a 15-A sphere of the ternary complex centered on the catalytic histidine. The search is carried out by calculating a 30-ps dynamics trajectory at 300 K and minimizing structures at 1-ps intervals. The protocol has been performed on 14 systems containing different combinations of substrate and mutant/wt LDH. In order to discover which interactions are important in defining substrate specificity, eight conformational parameters representing substrate-active site interactions were measured in each of the 420 minimized structures. These parameters were then compared to the measured catalytic activity of the protein-substrate combinations. These comparisons show that arginine 109 orientation is a major determining factor in LDH specificity. Using this methodology it is possible to estimate the catalytic activity of proteins of varied sequence by computer simulation before synthesis. PMID- 9329088 TI - Prediction of protein conformational freedom from distance constraints. AB - A method is presented that generates random protein structures that fulfil a set of upper and lower interatomic distance limits. These limits depend on distances measured in experimental structures and the strength of the interatomic interaction. Structural differences between generated structures are similar to those obtained from experiment and from MD simulation. Although detailed aspects of dynamical mechanisms are not covered and the extent of variations are only estimated in a relative sense, applications to an IgG-binding domain, an SH3 binding domain, HPr, calmodulin, and lysozyme are presented which illustrate the use of the method as a fast and simple way to predict structural variability in proteins. The method may be used to support the design of mutants, when structural fluctuations for a large number of mutants are to be screened. The results suggest that motional freedom in proteins is ruled largely by a set of simple geometric constraints. PMID- 9329089 TI - SAmBA: an interactive software for optimizing the design of biological macromolecules crystallization experiments. AB - SAmBA is a new software for the design of minimal experimental protocols using the notion of orthogonal arrays of strength 2. The main application of SAmBA is the search of protein crystallization conditions. Given a user input defining the relevant effectors/variables (e.g., pH, temperature, salts) and states (e.g., pH: 5, 6, 7 and 8), this software proposes an optimal set of experiments in which all tested variables and the pairwise interactions between them are symmetrically sampled. No a priori restrictions on the number and range of experimental variables is imposed. SAmBA consists of two complementary programs, SAm and BA, using a simulated annealing approach and a backtracking algorithm, respectively. The software is freely available as C code or as an interactive JAVA applet at http:/(/)igs-server.cnrs-mrs.fr. PMID- 9329103 TI - The relationship between the World Trade Organisation and the Office International des Epizooties. AB - The provisions of the World Trade Organisation Agreement on the Application of Sanitary and Phytosanitary Measures are designed to extend the liberalisation of trade, without increasing the risk to public, animal or plant health. The international standards set by the Office International des Epizooties (OIE) will be used as a benchmark by World Trade Organisation panels and committees when evaluating national sanitary-based regulations. For a significant liberalisation of trade to be achieved, Member Countries are faced with a dual mandate: a) each country must put these concepts into practice when making import/export decisions; and b) each country must make the commitment to support the OIE in its efforts to develop and review sanitary standards. Of equal importance to the application of standards is the cultural change that trade and regulatory communities must undergo. The author examines the role of Member Countries and the OIE in the implementation of this important agreement. PMID- 9329104 TI - Risk analysis in relation to the importation and exportation of animal products. AB - The design of a quantitative risk analysis model has to be dictated by the questions it seeks to answer. The model should also be as objective as the available data will allow. Animal and animal product import risks usually have three characteristics which make the design of a good quantitative risk analysis model quite difficult, namely:--the probabilities of the steps leading to the undesired outcome are frequently inter-related--the probability of the undesired outcome itself is in many cases very small, making direct simulation impractical- important variables within the model often cannot be quantified through analysis of data, thus these variables must be modelled with probability distributions to reflect the degree of uncertainty, usually determined by expert opinion. This paper provides a tutorial on some modelling techniques which are essential to the risk assessment of animal and animal product imports and which help overcome these problems. A number of probability distributions, their uses and inter relationships, are examined. The application of these distributions, coupled with some general modelling techniques, is then demonstrated to produce rigorous and transparent animal import risk analyses. PMID- 9329105 TI - Contamination of animal products: the minimum pathogen dose required to initiate infection. AB - When an animal product contains a low level of contamination (perhaps less than the minimum infective dose of a pathogen as determined experimentally), the theoretical probability remains that if a large number of animals are exposed to that product, at least one animal in the group will become infected. Such an infected animal could start an outbreak of the disease. These aspects, therefore, should be considered when risk assessments are performed. Foot and mouth disease virus in milk is used as an example. PMID- 9329106 TI - Risks of introducing foot and mouth disease through the importation of beef from South America. AB - The safety of beef with respect to foot and mouth disease (FMD) is determined by the level of risk which the exporting region poses through disease prevalence, the reliability of the surveillance system of the region, the efficacy of the prevention and control measures, the efficiency of the Veterinary Services and the support of the private sector. The South American continent has been regionalised in accordance with these criteria. Today there are approximately 90 million cattle in a territory of over 5 million km2 comprising regions classified as having a very low to low level risk for FMD with regard to the export of animals and animal products. Another 50 million cattle live in regions classified as posing a moderate risk. These risk categories reflect varying levels of risk. The harvest of beef in the meat-exporting regions of South America includes a series of risk mitigation measures, from the origin of the source herd to the final packing of the beef. These measures reduce the unrestricted risk estimate by almost six orders of magnitude. Therefore, the final risk of FMD for the global trade of beef originating from the low risk regions in South America is extremely small. PMID- 9329107 TI - The potential risks to animal health from imported sheep and goat meat. AB - Alerted by outbreaks of foot and mouth disease and the swine fevers which have been attributed to international trade in meat, regulators have tended to adopt conservative policies with respect to the importation of meat. However, for disease introduction to occur as a result of meat importation, a number of criteria must be satisfied. A qualitative assessment of the risks posed by sheep and goat meat leads to the conclusion that, with the possible exception of foot and mouth disease there is little likelihood that Office International des Epizooties List A or List B diseases would be spread through trade in adequately matured meat obtained from animals which have passed veterinary ante-mortem and post-mortem inspections. PMID- 9329108 TI - Health hazards to the small ruminant population of the Middle East posed by the trade of sheep and goat meat. AB - Meat consumers in the Middle East traditionally prefer meat from freshly slaughtered animals to that from chilled or frozen carcasses. Consequently, meat trade in the Middle East is based mainly upon the importation of large quantities of live animals rather than of sheep and goat carcasses. Furthermore, as it seems that pathogens remain viable for longer periods of time in live animals than in meat, the probability of pathogens spreading in the Middle East as a result of contaminated small ruminant carcasses is lower than the probability of pathogens being imported through live animals. With suitable environmental conditions, however, there are two livestock diseases which may spread through meat, namely: foot and mouth disease (FMD) and transmissible spongiform encephalopathies (TSEs). Foot and mouth disease virus which might have survived in the bones and offal of animals slaughtered in FMD-infected areas may be transmitted to animals in other regions. This occurs if carcass waste matter is not properly disposed of and is a particular problem where scavenging animals may carry away the infective material. Due to the low standards in garbage and sewage collection methods in many regions of the Middle East, such an eventuality should never be overlooked. Transmissible spongiform encephalopathies may be introduced into sensitive animal populations if the waste matter of slaughtered sheep and goats is processed into animal feed. This would occur if animal products which have not undergone treatment to inactivate the scrapie agent are used in the rendering process. The risk of introducing TSEs into livestock populations of the Middle East in this way is very low, since the rendering technology and the use of animal waste matter as feed are not part of the local traditional husbandry practices. PMID- 9329109 TI - Potential animal health hazards of pork and pork products. AB - The animal health hazards associated with the importation of pork and pork products include four viral agents: foot and mouth disease, classical swine fever (hog cholera), African swine fever, and swine vesicular disease viruses. The safety of importing pork from a zone infected with one or more of these diseases can be adequately determined only through risk assessment. This also applies for the safety of importing pork products which have undergone some form of processing (fully cooked pork products are not counted here). For each disease, the agent (pH and temperature lability), target organs, agent survival in pork and pork products, and agent quantification are discussed. Agent quantification is an input of the risk assessment which measures the viral titres in waste pork and pork products in relation to the oral infective dose estimated for each disease. Two other viral diseases, transmissible gastroenteritis of pigs and porcine reproductive and respiratory syndrome, are presented to illustrate why these two diseases are not hazards when associated with pork and pork products. PMID- 9329110 TI - Animal health risks associated with the transportation and utilisation of wildlife products. AB - The animal health risks associated with the movement of wildlife products are infinitely less than those associated with the movement of live animals. Very few pathogens are sufficiently robust to survive the significant changes in temperature, pH, moisture content and osmolality which occur post mortem, or which are associated with preservation processes such as pickling, smoking or drying. Certain pathogens, however, (e.g. foot and mouth disease, classical swine fever [hog cholera] and African swine fever viruses and the anthrax bacillus) are hardy and resistant to these environmental changes and therefore constitute a finite animal health risk if raw, undercooked or under-preserved products from infected wild animals are imported. Other less robust pathogens, such as rinderpest virus, may remain infectious in animal products if these are obtained from acutely infected animals and frozen immediately. Macroparasitic diseases such as trichinellosis and echinococcosis-hydatidosis, if present in the unprocessed tissues of infected wildlife, are potentially infectious to carnivorous or omnivorous companion animals. The importation of untreated wet hides may result in the introduction of alien ectoparasites and/or the infectious diseases for which they are vectors. The author discusses the more significant pathogens found in free-ranging wildlife which should be taken into consideration when importing wildlife products from endemically or epidemically infected countries. PMID- 9329112 TI - Risks of spreading foot and mouth disease through milk and dairy products. AB - A review of epidemics of foot and mouth disease (FMD) has highlighted the important role which raw (untreated) milk can play in the spread of the disease in a country which is normally free of FMD and whose cattle are not routinely vaccinated. The greatest hazard is likely to be in the early stages of an outbreak, before disease control measures have been implemented. The spread of FMD through milk can be prevented by the effective application of control measures combined with 'codes of practice' for the treatment of potentially infected milk. The author considers the probable mechanisms of transmission of FMD by milk and dairy products. These mechanisms are based on the quantities of virus excreted in milk, the survival of the virus under various management and manufacturing conditions and the minimum doses required to initiate infection in susceptible animals by different routes. The key points for consideration when making a risk assessment of the importation of milk and dairy products are also discussed. PMID- 9329111 TI - Animal health risks associated with ostrich products. AB - Five diseases recorded in ostriches are regarded as posing a potential animal health threat to meat-importing countries. Newcastle disease causes an atypically low mortality in ostriches: infected birds display typical nervous symptoms but no pathognomonic lesions which could be detected during post-mortem inspection. The vaccination of feedlot birds and a thorough ante-mortem examination are regarded as necessary precautions to ensure virus carriers are not among those animals destined for slaughter and subsequent export. Avian influenza produces clinical depression and lesions can be detected at post-mortem examination. Borna disease appears to affect mainly younger birds, and the virus is probably not present in the meat of affected birds. Finally, there is little evidence to suggest that ostriches could play a role in the epidemiology of transmissible spongiform encephalopathies. Cases of anthrax are extremely rare. The importation of deboned ostrich meat reduces the risk of infected scraps being fed to susceptible animals. PMID- 9329113 TI - Contamination of fish products: risks and prevention. AB - The risks of contamination of finfish products with active pathogens largely depend on the type of product concerned and disposal methods of the importing country. Frozen fish used as bait or to feed high-value species present the greatest risk as vehicles of contamination because they are unprocessed. Freezing preserves viral- and some bacterial-pathogens, thus the use of such fish as bait can introduce those pathogens into natural waters. Conversely, processed fish, particularly fillets, which have been heat-treated or cooked, present the lowest risk. If fish are processed after importation, care must be taken to ensure effective waste disposal, with particular attention to the prevention of scavenging by avian vectors and drainage from landfills into natural waters. Liquid waste should be disinfected and disposed of well away from natural waters. PMID- 9329114 TI - Risk of spread of penaeid shrimp viruses in the Americas by the international movement of live and frozen shrimp. AB - Within the past decade, viral diseases have emerged as serious economic impediments to successful shrimp farming in many of the shrimp-farming countries of the world. In the western hemisphere, the viral agents of Taura syndrome (TS) and infectious hypodermal and haematopoietic necrosis have caused serious disease epizootics throughout the shrimp-growing regions of the Americas and Hawaii, while in Asia the viral agents of white spot syndrome (WSS) and yellow head (YH) have caused pandemics with catastrophic losses. The international transfer of live shrimp for aquaculture purposes is an obvious mechanism by which the viruses have spread within and between regions in which they have occurred. Shrimp-eating gulls, other seabirds and aquatic insects may also be factors in the spread of shrimp viruses between and within regions. Another potentially important mechanism for the international spread of these pathogens is the trade in frozen commodity shrimp, which may contain viruses exotic to the importing countries. The viral agents of WSS, YH and TS have been found, and demonstrated to be infectious, in frozen shrimp imported into the United States market. Mechanisms identified for the potential transfer of virus in imported frozen products to domestic populations of cultured or wild penaeid shrimp stocks include: the release of untreated liquid or solid wastes from shrimp importing and processing plants directly into coastal waters, improper disposal of solid waste from shrimp importing and processing plants in landfills so that the waste is accessible to gulls and other seabirds, and the use of imported shrimp as bait by sports fishermen. PMID- 9329116 TI - Bovine spongiform encephalopathy: the causal role of ruminant-derived protein in cattle diets. AB - Although bovine spongiform encephalopathy (BSE) has occurred in other European countries, the major epidemic has been in the United Kingdom (UK), where there have been more than 163,000 cases so far. BSE has been linked to the practice of feeding meat-and-bone meal (MBM), putatively contaminated with scrapie agent, to cattle. A ban on the feeding of MBM to ruminants in the UK has resulted in a significant decline in the number of reported cases. It is considered that BSE in other European countries probably originated through the use of British MBM in the diets of cattle in these affected countries. Recently, in the UK, a new variant form of Creutzfeldt-Jakob disease in humans has been discovered, which does not appear to have occurred before the advent of BSE. It may have been caused by BSE agent, possibly as a consequence of dietary exposure. The use of MBM in the diets of any livestock species has now been prohibited in the UK. PMID- 9329115 TI - Bee health and international trade. AB - The international trade in bee products is a complex issue as a result of the diverse uses of these products. This is especially true with regard to honey. In most cases, honey is imported for human consumption: the high purchase and shipping costs preclude the use of honey as feed for bees. For these reasons, the risk of transmitting disease through honey is minimal. However, this risk should not be ignored, especially in those countries where American foulbrood is not known to occur. The importation of pollen for bee feed poses a definite risk, especially since there are no acceptable procedures for determining whether pollen is free from pathogens, insects and mites. Routine drying of pollen would reduce the survival of mites and insects, but would not have any impact on bacterial spores. Phytosanitary certificates should be required for the importation of honey and pollen when destined for bee feed. The declaration on the phytosanitary certificate should include country of origin, and should state whether the following bee diseases and parasitic mites are present: American foulbrood disease, European foulbrood disease, chalkbrood disease, Varroa jacobsoni and Tropilaelaps clareae. PMID- 9329117 TI - Likelihood of introducing selected exotic diseases to domestic swine in the continental United States of America through uncooked swill. AB - To help policy makers determine the need for current regulations (which require cooking of swill prior to feeding to swine), an assessment of the likelihood of exposing domestic swine in the continental United States of America (USA) to selected foreign animal disease agents by feeding uncooked swill was carried out. The hazard was assumed to originate from contraband food items entering the USA and subsequently being discarded in household waste. Such food waste may be collected by licensed waste feeders and fed to swine. This study showed that, of the four diseases studied, the probability of exposure was highest for the classical swine fever (hog cholera) virus. The median annual likelihood of one or more contaminated loads of swill being fed to swine in the continental USA was estimated as follows: classical swine fever virus: 0.063, foot and mouth disease virus: 0.043, swine vesicular disease virus: 0.005, African swine fever virus: 0.005. PMID- 9329118 TI - Risks and economic consequences of introducing classical swine fever into The Netherlands by feeding swill to swine. AB - An effective animal disease prevention and eradication programme is of great importance for meat-exporting countries such as the Netherlands. If a serious outbreak of disease were to occur, the eradication measures required by the European Union and a possible ban on meat exports would have severe economic consequences. However, historical and experimental information on which these programmes can be based is scarce. Furthermore, until recently, an integrated approach which combined the various aspects of outbreaks and risks with economic consequences was lacking. This paper describes a project based on such an integrated approach. The project covered the elicitation of expert knowledge and the development of the virus introduction risk simulation model (VIRiS). VIRiS integrates objective and subjective information concerning risks and consequences of virus introduction, and thus presents policy-makers with a useful tool for the evaluation of existing prevention programmes and possible alternatives. VIRiS is illustrated for classical swine fever. A comparison is made between the current situation and a hypothetical situation where the risk factor 'swill feeding' is completely eliminated. PMID- 9329119 TI - Disease risks to animal health from artificial insemination with bovine semen. AB - Two of the major goals of artificial insemination of domesticated animals are to achieve continuous genetic improvement and to prevent or eliminate venereal disease. In comparison with natural service, fewer males are needed to artificially inseminate the same number of females and to produce the same number of offspring. However, there are risks associated with artificial insemination, which has the potential to disseminate genetic defects and also to spread infectious disease nationally and internationally. This paper focuses on the risk of six specific diseases which are transmitted in bull semen and outlines the appropriate measures to prevent these risks. PMID- 9329120 TI - The risks of disease transmission by embryo transfer in cattle. AB - Guidelines for the safe international movement of livestock embryos are provided in the International Animal Health Code of the Office International des Epizooties, and recommendations for embryo processing, based on numerous research papers on embryo-pathogen interaction studies, are given in the Manual of the International Embryo Transfer Society. Risk assessment is the logical extension of these approaches, since it provides veterinary authorities with a complete package of information on which to base their import/export decisions. Risk assessment includes evaluation of disease prevalence, effectiveness of Veterinary Services and competence of the embryo collection team. It also takes account of the epidemiology and pathogenesis of the disease concerned. The application of risk assessment for embryo movement is illustrated in this paper by comparisons of the probabilities of transmitting foot and mouth disease, bluetongue and vesicular stomatitis by bovine embryos. The risk scenario pathway was divided into three phases for analysis. The first phase deals with the potential for embryo contamination, which depends on the disease situation in the exporting region, the health status of donor herds and donor cows, and on the pathogenetic properties of the disease agent. The second phase covers risk mitigation by use of the internationally accepted standards for embryo processing, and the third phase considers the risk reductions resulting from post-collection surveillance of donors and donor herds, and also from testing of embryo-collection (flushing) fluids for the disease agent. It was evident from this assessment that low risks of transmitting disease by international movement of bovine embryos depend initially on a low disease incidence in the exporting region and on easily recognisable disease signs. Competent embryo processing was also of great importance, and in the case of bluetongue, vector ecology had a major influence. In addition to providing a logical basis for import/export decisions, risk assessment is useful for evaluating the potential outcome of new research and for assessing the safety of the movement of embryos of other species for which little or no research information is available on embryo-pathogen interactions. PMID- 9329121 TI - Risks of transmitting scrapie and bovine spongiform encephalopathy by semen and embryos. AB - This paper reviews current knowledge on transmission of scrapie and bovine spongiform encephalopathy (BSE) by semen and embryos. In sheep, in particular, it is difficult to distinguish between the genetic transmission of susceptibility to scrapie and vertical transmission of the infection. Nevertheless, there is evidence that vertical transmission of infection does occur, probably across the placenta, but none to suggest a significant scrapie risk from semen. Two teams have studied scrapie transmission from experimentally infected sheep using embryo transfer. Whereas one team found no evidence for transmission, the results from the other team suggest that embryos, even after washing, might carry the disease into the offspring. In regard to goats, although genetic differences in susceptibility exist, they are much less obvious than in sheep. There is no evidence for vertical transmission or for transmission through semen and embryos. With regard to BSE, although it appears that genetic differences in susceptibility are absent or unimportant, some recent work does suggest that the disease may be passed from cow to calf. The route of transmission and stage or stages when this takes place are unclear, however. In conclusion, despite growing evidence to indicate that scrapie and BSE are unlikely to be transmitted through semen and embryos, more research is needed to confirm this. Furthermore, until all possibility of risk is ruled out, risk reduction methods must be considered, especially when semen and embryos are being imported into countries where the diseases do not exist. PMID- 9329122 TI - Risk analysis and international trade principles applied to the importation into Canada of caprine embryos from South Africa. AB - Between November 1994 and February 1995 over nine thousand Boer goat embryos were imported into Canada from the Republic of South Africa. This substantial international movement of animal genetics via embryos was achieved through the application of the risk analysis principles prescribed in Section 1.4. of the International Animal Health Code of the Office International des Epizooties (OIE). Integral to the development of the health certification procedures was the application of the fundamental principles of non-discrimination, harmonisation, equivalence and transparency defined in the World Trade Organisation Agreement on Sanitary and Phytosanitary measures. Risk mitigation interventions were founded upon full consideration of the potential for disease transmission by animal embryos as espoused by the International Embryo Transfer Society and the relevant standards contained in Appendix 4.2.3.3. of the OIE International Animal Health Code. All the embryos imported into Canada were implanted into synchronised recipients on arrival. Twenty months later there has been no evidence of disease in either the recipient animals or the resulting animals born in Canada. PMID- 9329123 TI - Hypertonic saline resuscitation: a tool to modulate immune function in trauma patients? AB - Hypertonic saline (HS) resuscitation has recently gained attention from trauma physicians because it may benefit the immune system of trauma patients. We have found that HS augments in vitro and in vivo immune function of healthy T-cells. In addition, HS restored the function of suppressed T-cells in vitro and in vivo and reduced immunosuppression after hemorrhage, protecting mice from subsequent sepsis. These effects of HS are based on its direct influence on cellular signaling events through specific signaling pathway(s) that include protein tyrosine kinase and mitogen-activated protein kinase p38 activation. HS provides a costimulatory signal that enhances the proliferation of activated T-cells. HS may be able to substitute signals lost through blockage as a result of trauma induced suppressive factors, thereby restoring the function of suppressed T cells. Although further work is needed to determine the optimal conditions and possible risks of HS resuscitation, the data presented in this short review of our recent work shed a favorable light on HS as a simple but effective tool to modulate cellular immune function after trauma. PMID- 9329124 TI - Flutamide: a novel agent for restoring the depressed cell-mediated immunity following soft-tissue trauma and hemorrhagic shock. AB - Recent studies indicate beneficial effects of androgen depletion in male mice, before trauma-hemorrhage on cell-mediated immunity following soft-tissue trauma and hemorrhagic shock. Nonetheless, it remains unknown whether androgen receptor blockade following the insult has any salutary effects. To study this, male C3H/HeN mice were either sham-operated or subjected to soft-tissue trauma (i.e., 2.5 cm midline laparotomy) followed by hemorrhagic shock (blood pressure 35 +/- 5 mmHg for 90 min) and then adequately resuscitated (shed blood and lactated Ringer's). Immediately after the completion of resuscitation, as well as 24 and 48 h thereafter, the animals received either vehicle, 10 mg/kg body weight (BW) flutamide or 25 mg/kg BW flutamide subcutaneously. At 72 h after resuscitation, all animals were killed. The spleens and peritoneal macrophages (M phi) were then harvested and cultures established to determine IL-2 and IL-3 release, splenocyte proliferative capacity, as well as splenic and peritoneal M phi IL-1 release. Moreover, plasma testosterone and corticosterone levels were measured. Our results indicate that trauma-hemorrhage resulted in significant depression of splenocyte and M phi functions in vehicle-treated and animals receiving 10 mg/kg BW flutamide. Treatment with 25 mg/kg BW flutamide following trauma-hemorrhage, however, resulted in levels of cytokine release which were comparable with those found in sham-operated animals. No significant alterations in plasma corticosterone and testosterone levels were observed in any of the experimental groups. These findings indicate that short-term therapy of males with the androgen receptor blocker, flutamide at 25 mg/kg BW, following trauma-hemorrhage has protective effects on immune functions. This protective effect is dose dependent, since 10 mg/kg BW flutamide did not produce significant salutary effects. Thus, flutamide represents a novel and safe agent for improving the depressed functions in male trauma patients suffering severe blood loss. PMID- 9329125 TI - Interleukin-1 beta and interferon-gamma regulate interleukin-6 production in cultured human intestinal epithelial cells. AB - Recent studies suggest that interleukin-6 (IL-6) is produced in the intestinal mucosa during sepsis and endotoxemia and that the enterocyte may be a source of IL-6 in these conditions. The regulation of IL-6 production in the enterocyte is not fully understood. We tested the hypothesis that IL-6 production in the enterocyte is regulated by proinflammatory cytokines. This was done by treating cultured Caco-2 cells, a transformed human intestinal epithelial cell line, with different concentrations of tumor necrosis factor-alpha (TNF-alpha), IL-1 beta, IL-6 or interferon-gamma (IFN-gamma). IL-6 production by the Caco-2 cells was determined by ELISA. The expression of IL-6 mRNA was determined by reverse transcriptase polymerase chain reaction. IL-6 was not produced in unstimulated Caco-2 cells. Treatment of the Caco-2 cells with IL-1 beta resulted in a dose- and time-dependent stimulation of IL-6 production with a maximal effect noted at an IL-1 beta concentration of .5 ng/mL at 24 h. IFN-gamma alone did not stimulate IL-6 production but potentiated the effect of IL-1 beta in a synergistic fashion. Treatment of the Caco-2 cells with IL-1 beta induced expression of IL-6 mRNA with a response noticed after 30 min. TNF-alpha and IL-6 did not influence the production of IL-6 in the Caco-2 cells. The results suggest that enterocyte IL-6 production is stimulated by IL-1 beta and that this effect is potentiated by IFN gamma. The regulation of IL-6 production in the enterocyte may be specific for IL 1 beta, since neither TNF nor IL-6 stimulated IL-6 production. PMID- 9329126 TI - Elevated expression of p47phox and p67phox proteins in neutrophils from burned rats. AB - The molecular control of neutrophil respiratory burst in burn injury was investigated through quantitation of protein factors, p47phox and p67phox, which are required for the activation of the phagocyte plasma membrane NADPH-oxidase. Circulating neutrophils were isolated from rats with 30% body surface area covered with full thickness burns. Neutrophil O2- generation, and p47phox and p67phox expressions, respectively, were determined using spectrophotometric and immunoblot techniques. Formylmethionyl-leucyl-phenylalanine stimulated superoxide anion generation was approximately 50% higher in neutrophils from rats 24 and 72 h after burns compared with that in sham control rats. The level of superoxide production was .47 +/- .05 nanomoles per minute per 10(6) cells (mean +/- SE, n = 6) at 24 h and .45 +/- .05 (n = 6) at 72 h postburn, whereas in sham control animals it was .32 +/- .02 (n = 8). Compared with the sham group p47phox levels, p47phox expression was 5.7-fold, 4.4-fold, and 4.5-fold higher, respectively, at 24, 36 and 72 h postburn. The levels of p67phox in burned animals were 2-fold higher than in the sham group, (p < .05) at 24 h postburn, and approximately 50% higher than sham at 36 h after the burn. The p67phox levels in rats 72 h after the burn were not significantly different from the sham values. These data support the occurrence of an up-regulation of p47phox and p67phox expressions accompanying the enhanced neutrophil respiratory burst activity during the early stages of burn injury. The up-regulation of p47phox and p67phox could be responsible for the priming of neutrophil O2- production leading to host tissue injury. PMID- 9329127 TI - Gradual development of organ damage in the murine zymosan-induced multiple organ dysfunction syndrome. AB - A well defined animal model is a prerequisite to test intervention strategies aimed at preventing the development of the multiple organ dysfunction syndrome. This study compares changes in clinical parameters to histopathologic changes in tissues, observed over a 12 day period after a single intraperitoneal injection of zymosan in C57BL/6 mice. Administration of zymosan induced gradual and progressive changes in wet and dry organ weight of the liver, kidneys, and particularly, lungs and spleen that correlated with increasing histopathology. From 6 days after zymosan injection onwards, hemorrhagic spots were found in the lungs evolving into massive hemorrhages at 12 days, when thickened interstitial walls and loss of the honey comb-like structures were observed. The liver displayed progressive accumulation of macrophage-like and mononuclear cells. After 12 days, numerous granuloma-like structures were disseminated throughout the liver parenchyma. The spleen displayed great changes in red and white pulp with increasing numbers of megakaryocytes and plasma-like cells. In the kidneys, necrosis of the tubular epithelium adjacent to granulomas on the ventral (peritoneal) side was found. In mice, a single intraperitoneal challenge with zymosan leads to progressive multiple organ damage, which becomes apparent at some time after the insult. This animal model displays a number of features encountered in human multiple organ dysfunction syndrome and appears suitable to conduct intervention studies. PMID- 9329128 TI - Multiple organ failure following zymosan-induced peritonitis is mediated by nitric oxide. AB - In the present study we tested the hypothesis that nitric oxide may play a role in the pathogenesis of multiple organ failure induced by peritoneal injection of zymosan in the rat. A severe inflammatory response characterized by peritoneal exudation, high plasma and peritoneal levels of nitrate/ nitrite (breakdown products of nitric oxide), prostaglandin E2 and leukocyte infiltration into peritoneal exudate was induced by zymosan administration. This inflammatory process started within 3 h of administration and onset occurred at 18 h, coinciding with damage of lung, small intestine and liver, as assessed by histological examination and by increase of myeloperoxidase activity, indicative of neutrophil infiltration. Furthermore, at 18 h after zymosan-induced peritonitis, expression of inducible nitric oxide synthase enzyme was found mainly in the macrophages of inflamed lungs. Subcutaneously administration of a nonisoform selective nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester, reduced formation of peritoneal exudate fluid, blocked plasma and peritoneal nitrate/nitrite accumulation, and attenuated the elevated release of peritoneal prostaglandin E2. In addition, nitric oxide synthase inhibition was effective in preventing the development of organ failure since tissue injury and neutrophil infiltration, by myeloperoxidase evaluation, was reduced in lung, small intestine, and liver. In conclusion, major findings of our study are that nitric oxide exerts a proinflammatory role in the development of multiple organ failure and nitric oxide synthase inhibition is an effective antiinflammatory therapeutic tool, since inhibits not only nitric oxide but also prostaglandin production and cellular infiltration in inflamed organs. PMID- 9329129 TI - Differences in eicosanoid and cytokine production between injury/hemorrhage and bacteremic shock in the pig. AB - Plasma concentrations of the eicosanoids leukotriene (LT)B4, LTC4D4E4, thromboxane (TX)A2 and prostaglandin (PG)I2, and tumor necrosis factor (TNF) were measured during acute bacteremic shock and injury/hemorrhage in two porcine models. As TXA2 and PGI2 are rapidly metabolized, we measured their stable metabolites TXB2 and 6-keto-PGF1 alpha. Bacteremic shock was induced by a graded infusion of Aeromonas hydrophila over 4 h. Injury/hemorrhage was produced by a 30 min, 30% total blood volume hemorrhage followed by a 30 min shock period and then reinfusion of shed blood. Nociceptive afferent nerve stimulation was applied to the brachial plexi to mimic the cardiovascular responses to tissue injury. There was no increase in eicosanoid or TNF levels in the injury/hemorrhage model. In sepsis there was an early peak in TNF (at 60 min) followed by peaks in LTB4 and LTC4D4E4 at 180 min. Both TXB2 and 6-keto-PGF1 alpha showed large increases at the end of the study but there was no evidence that they had reached a peak. These results suggest that the very early inflammatory response in bacteremic shock and injury/hemorrhagic shock may be quite different. This may have implications for any therapies aimed at reducing the incidence of multiple organ failure after either of these physiological insults. PMID- 9329131 TI - Resuscitation of pulmonary contusion: hypertonic saline is not beneficial. AB - We postulated that hypertonic solutions could minimize the accumulation of lung water and subsequent respiratory derangements that occur after pulmonary contusion. Anesthetized pigs underwent contusion of the right chest at baseline and then were hemorrhaged (30 cc/kg) over 20 min. They were resuscitated with either 7.5% NaCl (4 cc/kg) or .9% saline (90 cc/kg) for 20 min and observed for 4 h. Gravimetric lung weights and spiral computed tomography scans were used to quantitate lung water. The hemodynamic response to contusion and hemorrhage was similar in both resuscitation groups. Arterial oxygen tension was not significantly altered by the method of resuscitation and remained close to baseline values for the entirety of the experiment. Static compliance measurements were significantly decreased from baseline in both groups following pulmonary contusion. There were no differences in wet to dry lung weights or computed tomography scan injury volume between groups. We conclude that small volume hypertonic saline resuscitation does not reduce the magnitude of lung injury or provide substantial physiologic benefit over isotonic solutions following pulmonary contusion. PMID- 9329130 TI - The role of mast cells in mucosal permeability changes during ischemia reperfusion injury of the small intestine. AB - The objective of this study was to investigate the significance of mast cell induced reactions in the mucosal functional and morphological alterations induced by 30 min segmental ischemia and 120 min reperfusion in anesthetized dogs. The rates of changes in permeability of the mucosa to sodium fluorescein (NaFL) in the plasma-to-lumen and lumen-to-plasma directions were studied, the local hemodynamics, intramucosal pH (pHi) alterations, mast cell number and degranulation, and degree of tissue injury were determined. The effects of pretreatments with cromolyn (a peritoneal-type mast cell stabilizer), quercetin (a mucosal-type mast cell stabilizer), and dexamethasone (an aspecific membrane stabilizer and mast cell depleter) were evaluated. We found that ischemia reperfusion induced significant tissue injury, elevated the segmental vascular resistance, and decreased pHi. The plasma-to-lumen clearance of NaFL increased significantly during ischemia and reperfusion. Cromolyn and quercetin pretreatments significantly inhibited the permeability changes, but did not influence the pHi and morphological alterations induced by ischemia-reperfusion. Dexamethasone pretreatment did not influence the number of mast cells, but the degree of mast cell degranulation and fluorescein leakage decreased. We conclude that intestinal mast cells and mast cell-induced reactions contribute to the mucosal permeability alterations during reperfusion, but play only a minor role in ischemia-reperfusion-induced structural injury. PMID- 9329132 TI - Xanthine oxidase inhibition after resuscitated hemorrhagic shock restores mesenteric blood flow without vasodilation. AB - To determine the contribution of xanthine oxidase-mediated reperfusion injury to the blood flow deficits seen in the intestinal microcirculation after resuscitated hemorrhagic shock, rats were prepared for intravital microscopic study then bled to 50% of baseline blood pressure for 60 min. Treatment animals received a 50 mg/kg bolus and a 25 mg/kg/h infusion of the xanthine oxidase inhibitor allopurinol after shock but before standard resuscitation with shed blood and an equal volume of Ringer's lactate. A similarly resuscitated group served as control. Blood flow and vessel diameters were measured in the neurovascularly intact terminal ileum using intravital microscopy and doppler velocimetry. Resuscitation restored cardiac output and blood pressure in both groups. Blood flow in first order arterioles 120 min postresuscitation was 41% of baseline in the standard resuscitation group and 77% of baseline in the allopurinol-treated group. A1 arteriolar diameter was not significantly different between the two groups, being 73 and 82% of baseline, respectively. These data suggest that xanthine oxidase-mediated ischemia-reperfusion injury contributes to blood flow deficits in the small intestinal microcirculation after resuscitated hemorrhagic shock and that the improvement in blood flow seen with allopurinol is not due to vasodilation within the microvasculature. PMID- 9329133 TI - The effects of Escherichia coli sepsis and short-term ischemia on coronary vascular reactivity and myocardial function. AB - Ischemia and reperfusion stun the myocardium and the coronary vasculature. We have previously shown that a short period (15 min) of global ischemia in the isolated rat heart causes impaired coronary constriction in response to a thromboxane analog U46619 during reperfusion. Sepsis has also been shown to affect myocardial and vascular function. In the present study, we determined whether Escherichia coli-induced sepsis would exacerbate the effects of ischemia on the coronary circulation. Sepsis prolonged the impairment in the coronary constriction response to U46619 following short term ischemia. We hypothesized that sepsis-induced increases in nitric oxide (NO) production caused the delay in the recovery of the contractile response to U46619. Perfusion with NO synthase inhibitors however indicated that the impaired response was not due to NO. However, NO did appear to have a significant role in the development of myocardial ischemic contracture and on the recovery of diastolic function after ischemia. Inhibitors of NO synthase also caused a significant increase in basal coronary perfusion pressure as well as in the maximum coronary pressure generated in response to U46619, suggesting a role of NO in regulating basal coronary vascular resistance in the isolated rat heart. Some of these effects were more pronounced in septic rat hearts than in the sham surgical rat hearts, consistent with altered nitric oxide production in the septic rat hearts. PMID- 9329151 TI - Role of glutamate receptor subtypes in the differential release of somatostatin, neuropeptide Y, and substance P in primary serum-free cultures of striatal neurons. AB - The spiny and aspiny neuronal populations of the striatum display differential vulnerability to the toxic effects of glutamatergic agonists. Substance P containing spiny neurons appear to be more vulnerable to NMDA-receptor-mediated toxicity and less susceptible to kainate toxicity than the somatostatin- and neuropeptide Y (NPY)-containing aspiny population. We studied whether selective glutamatergic agonists might have similar differential effects on neuropeptide release from the substance P- and somatostatin/NPY-containing neuronal populations. After collection of a baseline sample, striatal neurons in primary culture were treated with one of the following: phosphate-buffered saline, 56 mM potassium chloride (KCl), 100 microM N-methyl-D-aspartate (NMDA), 100 microM quisqualate, 100 microM kainate, or 100 microM glutamate. Baseline and treatment samples were measured by radioimmunoassay for somatostatin, NPY, and substance P. KCl and kainate provoked a selective release of somatostatin and NPY, whereas substance P measured in the same samples showed no response. By contrast, NMDA elicited a selective release of substance P without a similar increase of either somatostatin or NPY. Quisqualate evoked comparable responses in the three peptides. These results indicate that the glutamatergic regulation of somatostatin and NPY release from aspiny striatal neurons in primary culture is preferentially mediated by the kainate receptor, whereas substance P release is selectively mediated by the NMDA receptor. These findings suggest a preferential expression of functional kainate receptors on the aspiny somatostatin/NPY neurons and of NMDA receptors on the substance-P-containing spiny neurons. PMID- 9329152 TI - Glutamate receptors in the postmortem striatum of schizophrenic, suicide, and control brains. AB - INTRODUCTION: Previous postmortem studies of glutamate receptors and uptake sites have shown decreased D-aspartate (D-Asp) (a marker for the high affinity glutamate uptake site) and elevated (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine maleate (MK-801) binding in the putamen in schizophrenia and elevated alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor binding in the caudate nucleus of schizophrenics who committed suicide. The relative effects of schizophrenia, suicide, and neuroleptic treatment in these findings is unclear. This study further explores binding to glutamate receptors (NMDA, kainic acid, and AMPA) and uptake sites in postmortem striatal structures in schizophrenics relative to three control groups (normal controls, neuroleptic-treated controls, and nonpsychotic suicides). METHODS: We compared the binding densities of tritium-labeled ligands 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX), kainic acid (KA), MK-801, and D-Asp, which target the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), KA, and N methyl-D-aspartic acid (NMDA) ionotropic receptor sites and the glutamate uptake site, respectively, in postmortem striatal/accumbens tissue from six DSM-III-R schizophrenics, eight normal controls, eight neuroleptic-treated controls, and eight suicide victims using standard receptor autoradiographic methods. RESULTS: Binding of [3H] CNQX (AMPA receptors) was significantly different among the four groups across the subdivisions of the striatum: caudate, putamen, and nucleus accumbens (ANOVA P = .0007, .002, and .004, respectively). The schizophrenia group had higher mean CNQX binding in the caudate nucleus than normal (P = .005) and neuroleptic controls (P = .006) but not suicides (P = .6), who were also higher than normals and neuroleptic-treated controls (P = .05). The binding densities of tritiated MK-801, KA, and D-Asp were not significantly different among the four groups of subjects in any of the striatal regions examined. CONCLUSIONS: The data suggest there may be an increased density of AMPA receptor sites in the caudate nucleus in schizophrenia that is not apparently due to neuroleptic treatment. A similar increase was also seen the suicide group. Although these data do not confirm previous reports of an increase in [3H]MK-801 or a decrease in [3H]D-Asp binding in the basal ganglia in schizophrenia, the increased caudate AMPA binding observed here could reflect decreased cortical glutamatergic innervation of the caudate. Its implication for suicide is unclear. PMID- 9329153 TI - Effects of intracerebroventricular administration of beta-funaltrexamine on DAMGO stimulated [35S]GTP-gamma-S binding in rat brain sections. AB - Intracerebroventricular administration of beta-funaltrexamine (beta-FNA) reduces the density of mu opioid receptors as measured by in situ autoradiography by 40 50% throughout the brain, with little regional variation [Martin et al. (1993) J. Pharmacol. Exp. Ther. 267:506-514] Recently an assay has been developed to study opioid stimulation of [35S]GTP-gamma-S binding autoradiographically in situ using slide-mounted brain sections [Sim et al. (1995) Proc. Natl. Acad. Sci. U.S.A. 92:7242-7246]. The present study was undertaken to determine the effect of mu opioid receptor alkylation on G protein activation by the mu opioid agonist DAMGO. Animals were injected intracerebroventricularly with 40 nmol of beta-FNA or saline and sacrificed 24 hours later. DAMGO stimulated [35S]GTP-gamma-S binding with an anatomical specificity consistent with the localization of mu opioid receptors. The percent stimulation by DAMGO ranged from approximately 50 to 100% in the regions studied. beta-FNA significantly decreased G protein activation by DAMGO in regions that are consistent with its reported long-lasting and insurmountable antagonism of the antinociceptive (medial thalamus, central gray) and reinforcing (nucleus accumbens) effects of mu opioid agonists [Adams et al. (1990) J. Pharmacol. Exp. Ther. 255:1027-1032; Martin et al. (1995) J. Pharmacol. Exp. Ther. 272:1135-1140]. However, the effects of beta-FNA were not equal in all brain regions. This may indicate regional differences in the coupling efficiency of mu opioid receptors with G proteins, or in the effects of beta-FNA on mu opioid receptor binding or on mu opioid receptor-stimulated G protein activity. PMID- 9329154 TI - [123I]FP-CIT binds to the dopamine transporter as assessed by biodistribution studies in rats and SPECT studies in MPTP-lesioned monkeys. AB - [123I]FP-CIT (N-omega-fluoropropyl-2 beta-carbomethoxy-3 beta-(4-iodophenyl) tropane), a radioiodinated cocaine analogue, was evaluated as an agent for the in vivo labeling of dopamine (DA) transporters by biodistribution studies in rats and by single photon emission computed tomography (SPECT) studies in unilateral 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys. In rats, intravenous injection of [123I]FP-CIT resulted in high accumulation of radioactivity in the striatum. Less pronounced uptake was seen in brain areas with high densities of serotonergic uptake sites. While striatal uptake of radioactivity after injection of [123I]FP-CIT was displaced significantly by GBR12,909 but not by fluvoxamine, the opposite was observed in brain areas known to be rich of serotonin transporters. Monkeys which were unilaterally treated with neurotoxic doses of MPTP showed severe loss of striatal [123I]FP-CIT uptake at the side of treatment. The results of this study indicate that [123I]FP-CIT, although not being a selective radioligand, binds specifically to the striatal DA transporter in vivo and thus suggest that [123I]FP-CIT promises to be a suitable radioligand for SPECT imaging of DA transporters in humans. PMID- 9329155 TI - Extracellular sodium removal increases release of neuropeptide Y-like immunoreactivity from rat brain hypothalamic synaptosomes: involvement of intracellular acidification. AB - Rat hypothalamic synaptosomes were exposed via superfusion to various stimuli and the release of neuropeptide Y-like immunoreactivity (NPY-LI) was measured by means of radioimmunoassay procedures. High KCl (15-50 mM) concentration dependently evoked NPY-LI release; the evoked overflow reached a plateau at 30 mM KCl and was abolished in the absence of Ca2+ ions. Furthermore, a remarkable NPY LI overflow was obtained when extracellular Na+ ions were removed. Low external Na(+)-evoked NPY-LI release was independent of the presence of Ca2+ ions from the superfusion medium. It is well known that the reduction of external Na+ ions activates the release of several neurotransmitters through an inversion of the uptake-carrier working direction; but such mechanisms, involving Na(+)-dependent uptake, have never been described for neuropeptides. The alteration of the extracellular Na+ concentration is able to modify the concentration of the intracellular Ca2+ and H+ ions. In fact, the concentrations of these two ions are regulated through Na(+)-dependent exchange mechanisms across the membrane. Amiloride, blocking the Na+/H+ exchanger, was able to maintain low Na(+)-evoked NPY-LI release, underlying that the blockade of the exchanger preserves the H+ accumulation induced by the reduction of the external Na+ ions. NPY-LI release could also be stimulated by nigericine, a proton ionophore, showing that the intracellular acidification is responsible for NPY-LI release. Intracellular acidification may stimulate Ca2+ ion release from intracellular stores, as has been shown by other workers. Large dense-core vesicles containing the peptide appear to be more sensitive to local intracellular Ca2+ release compared with extracellular Ca2+ ion entry through voltage-dependent channels. PMID- 9329156 TI - Staurosporine but not chelerythrine inhibits regeneration in hippocampal organotypic cultures. AB - A mechanical lesion in hippocampal organotypic cultures is followed by a recovery process involving scar formation, sprouting of fibres and formation of new functional synapses. Here we tested the effect of staurosporine and chelerythrine, two protein kinase C (PKC) inhibitors, on this lesion-induced neurite outgrowth of Shaffer collaterals. At a concentration of 1 microM, staurosporine delayed functional recovery assessed by measuring synaptic field potentials across the lesion, without altering synaptic transmission on nonlesioned cultures. Immunostaining carried out by using antibodies directed against neurofilament proteins showed that there was a marked reduction in the number of regenerating fibres crossing the lesion. In contrast to this, chelerythrine (50 microM) did not prevent functional recovery, although it affected synaptic transmission and plasticity at this concentration. We conclude that the inhibition of sprouting produced by staurosporine is independent of its blockade of PKC-mediated phosphorylation mechanisms. PMID- 9329157 TI - Alzheimer disease hyperphosphorylated tau aggregates hydrophobically. AB - The chemical interaction that condenses the hyperphosphorylated protein tau in Alzheimer's disease (AD P-tau) into neurofibrillary tangles and cripples synaptic transmission remains unknown. Only beta-sheet, positive ion salt bridges between phosphates, and hydrophobic association can create tangles of just AD P-tau. We have correlated transmission electron microscope (TEM) images of tau aggregation with different percentages of beta-sheet in aqueous suspensions of tau while using buffers that block dispositive or tripositive ionic bridges between intermolecular phosphates. Circular dichroism (CD) studies were performed at different temperatures from 5-85 degrees C using AD P-tau, AD P-tau dephosphorylated with hydrofluoric acid (HF AD P-tau) or alkaline phosphatase (AP AD P-tau), and recombinant human tau with 3-repeats and two amino terminal inserts (R-39) and using bovine tau (B tau) isolated without heat or acid treatment. Secondary structure was estimated from CD spectra at 5 degrees C using the Lincomb algorithm. Each preparation except one demonstrated an inverse temperature transition, Ti, in the CD at 197 nm. No correlation was found between beta-sheet content and aggregation, leaving only hydrophobic interaction as the remaining possibility. Thirteen of 21 possible phosphorylation sites in AD P-tau lie adjacent to positive residues in tau's primary structure. Occupation of five to nine phosphate sites on AD P-tau appears sufficient to reduce or neutralize tau's basic character. AD P-tau's hydrophobic character is indicated by its low inverse temperature transition, Ti. The Ti for AD P-tau was 24.5 degrees C or 28 degrees C, whereas for B tau with three phosphates it was 32 degrees C, for unphosphorylated tau R-39 it was 38 degrees C, and for dephosphorylated HF AD P tau it was 37.5 degrees C. The hydrophobic protein elastin and its analogs coalesce and precipitate at their Ti of 24-29 degrees C, well below body temperature. We hypothesize that AD P-tau causes tangle accumulation by this mechanism. PMID- 9329159 TI - Brain microdialysis of GABA and glutamate: what does it signify? AB - Microdialysis has become a frequently used method to study extracellular levels of GABA and glutamate in the central nervous system. However, the fact that the major part of GABA and glutamate as measured by microdialysis does not fulfill the classical criteria for exocytotic release questions the vesicular origin of the amino acids in dialysates. Glial metabolism or reversal of the (re)uptake sites has been suggested to be responsible for the pool of nonexocytotically released amino-acid transmitters that seem to predominate over the neuronal exocytotic pool. The origin of extracellular GABA and glutamate levels and, as a consequence, the implications of changes in these levels upon manipulations are therefore obscure. This review critically analyzes what microdialysis data signify, i.e., whether amino-acid neurotransmitters sampled by microdialysis represent synaptic release, carrier-mediated release, or glial metabolism. The basal levels of GABA and glutamate are virtually tetrodotoxin- and calcium independent. Given the fact that evidence for nonexocytotic release mediated by reversal of the uptake sites as a release mechanism relevant for normal neurotransmission is so far limited to conditions of "excessive stimulation," basal levels most likely reflect a nonneuronal pool of amino acids. Extracellular GABA and glutamate concentrations can be enhanced by a wide variety of pharmacological and physiological manipulations. However, it is presently impossible to ascertain that the stimulated GABA and glutamate in dialysates are of neuronal origin. On the other hand, under certain stimulatory conditions, increases in amino-acid transmitters can be obtained in the presence of tetrodotoxin, again suggesting that aspecific factors not directly related to neurotransmission underlie these changes in extracellular levels. It is concluded that synaptic transmission of GABA and glutamate is strictly compartmentalized and as a result, these amino acids can hardly leak out of the synaptic cleft and reach the extracellular space where the dialysis probe samples. PMID- 9329158 TI - Distribution of dopamine beta-hydroxylase-like immunoreactive fibers within the shell subregion of the nucleus accumbens. AB - The nucleus accumbens (Acb) can be divided into distinct subfields, delineated on the basis of histochemical markers as well as by afferent and efferent projection patterns. The shell subregion has reciprocal relationships with a variety of limbic areas and brainstem autonomic structures, and has been suggested to participate in motivation-related processes, including reward, stress, and arousal. The locus coeruleus (LC)-noradrenergic system has similarly been implicated in the modulation of behavioral state and stress-related processes, and previous studies have demonstrated reciprocal projections between the locus coeruleus and Acb shell. To better understand the anatomical substrate through which LC could influence activity within Acb shell, immunohistochemical methods were used to visualize the extent and the distribution of noradrenergic axons within this structure. Coronal sections of rat brain were processed to visualize immunoreactivity for the norepinephrine synthetic enzyme dopamine beta hydroxylase (DBH), a specific marker for noradrenergic processes. In some cases, alternate sections were processed for immunohistochemical localization of substance P, in order to delineate core, shell, and pallidal compartments. Moderate-to-dense DBH-like immunoreactivity (DBHir) was found in approximately the caudal half of the shell subregion, particularly in caudalmost (septal pole) and ventral zones. The innervation of the septal pole was contiguous with a dense innervation of the bed nucleus of the stria terminalis. Few immunoreactive fibers were observed in the caudate-putamen, Acb core, or rostral Acb shell. Many DBHir fibers within the shell region were highly arborized with numerous varicosities, features indicative of terminal fields. These observations suggest noradrenergic systems might modulate certain processes associated with stress, behavioral state, or reinforcement via actions within the Acb shell. PMID- 9329160 TI - Striatal dopamine receptors in rats displaying long-term behavioural sensitization to morphine. PMID- 9329161 TI - Introduction: birth defects surveillance in the United States. PMID- 9329162 TI - Birth defect surveillance at the state and local level. PMID- 9329163 TI - State-by-state cost of birth defects--1992. PMID- 9329164 TI - Prevention of neural tube defects. PMID- 9329165 TI - An annotated bibliography of the National Birth Defects Prevention Network. PMID- 9329166 TI - State birth defects surveillance programs directory (updated July 1997). PMID- 9329175 TI - Synthesis and chiroptical properties of a novel C2-symmetric binaphthyl phosphortriamide ("chiral HMPA"). AB - By use of an asymmetric Ullmann coupling involving chiral naphthalene oxazolines 1, the title compounds were prepared in good yields and with high diastereoselectivity. Hydrolysis of the binaphthyl oxazolines 2 led to the di aldehydes 5, which were transformed into the azepine derivative, 6. The latter was treated with the appropriate phosphoryl halide to access the chiral HMPA systems 7 and 9. The CD spectra of the chiral azepine 6 and the chiral phosphoramides 7 and 9 were measured and showed a strong positive CD couplet near 225 nm, consistent with the P axial chirality (S configuration). Semi-empirical CNDO/S molecular orbital calculations of the CD spectrum of 6 satisfactorily reproduced the major features of the observed spectrum. PMID- 9329167 TI - Birth defects surveillance data from selected states. PMID- 9329176 TI - Synthesis and cataleptic effects of optically active dihydrohaloperidols and dihydrobromoperidols. AB - Optically active dihydrohaloperidols and dihydrobromoperidols, the major metabolites of haloperidol and bromoperidol, clinically used as neuroleptic drugs in humans, were asymmetrically synthesized by Jaen's method. The motor effects of all the reduced haloperidol and bromoperidol metabolites were evaluated by the mouse catalepsy test. The results indicate that administration of the optically active dihydrohaloperidols and dihydrobromoperidols as well as haloperidol and bromoperidol can cause significant motor deficits in mice. PMID- 9329177 TI - Synthetic and model computational studies of molar rotation additivity for interacting chiral centers: a reinvestigation of van't Hoff's principle. AB - When plane-polarized light impinges on a solution of optically active molecules, the polarization of the light that emerges is rotated. This simple phenomenon arises from the interaction of light with matter and is well understood, in principle, van't Hoff's rule of optical superposition correlates the molar rotation with the individual contributions to optical activity of isolated centers of asymmetry. This straightforward empirical additivity rule is rarely used for structure elucidation nowadays because of its limitations in the assessment of conformationally restricted or interacting chiral centers. However, additivity can be used successfully to assign the configuration of complex natural products such as hennoxazole A if appropriate synthetic partial structures are available. Therefore, van't Hoff's principle is a powerful stereochemical complement to natural products' total synthesis. The quest for reliable quantitative methods to calculate the angle of rotation a priori has been underway for a long time. Both classical and quantum methods for calculating molar rotation have been developed. Of particular practical importance for determining the absolute structure of molecules by calculation is the manner in which interactions between multiple chiral centers in a single molecule are included, leading to additive or non-additive optical rotation angles. This problem is addressed here using semi-empirical electronic structure models and the Rosenfeld equation. PMID- 9329178 TI - Synthesis of novel 3'-trifluoromethyl taxoids through effective kinetic resolution of racemic 4-CF3-beta-lactams with baccatins. AB - The coupling of racemic 1-tBoc-4-CF3-beta-lactams with various C-10 modified baccatins has resulted in CF3-taxoids with diastereoselectivities ranging from 9:1 to one single isomer. The observed high diastereoselectivity is ascribed to the highly efficient enantiomer-differentiation by the enantiopure lithium alkoxide of a baccatin III in the coupling reaction with a racemic 1-tBoc-beta lactam. These novel CF3-taxoids have also been shown to exhibit significant increases in activity against various cancer cell lines compared to either paclitaxel or docetaxel. In addition, the first asymmetric synthesis of a CF3 beta-lactam via chiral ester enolate-imine cyclocondensation was performed with 50% enantioselectivity. PMID- 9329179 TI - Direct resolution, characterization, and stereospecific binding properties of an atropisomeric 1,4-benzodiazepine. AB - The chromatographic resolution of 7-chloro-1,3-dihydro-1-(1,1-dimethylethyl)-5- (2-fluorophenyl)-2H-1,4-benzodiazepin-2-on (7), the 2'-fluoro, N1-tertbutyl analogue of diazepam, was attained on both analytical and preparative (mgs) scales, by using several chiral stationary phases (CSPs). The stereochemistry of this compound was characterized by means of 1H-NMR Nuclear Overhauser Effect (NOE) analysis. The single enantiomers of 7 were tested for their configuration and stereochemical stability by circular dichroism (CD), and their interaction with the central nervous system (CNS) benzodiazepine receptor was assayed, showing a significant difference in their binding affinities. Protein binding studies with human serum albumin (HSA, the main benzodiazepine carrier in human plasma) immobilized on a silica stationary phase revealed that HSA also preferentially binds one stereoisomer of 7. However, both on line CD detection and stereospecific interaction with other common drugs clearly demonstrated that the stereoselectivity of immobilized HSA for 7 is opposite to that for all the other studied benzodiazepines. In addition, HSA stereoselectivity for 7 is opposite to CNS receptor binding stereoselectivity for the same compound. Such HSA anomalous stereoselectivity for 7 was also confirmed in aqueous buffer solution by competitive displacement studies. Compared to other chiral 1,4 benzodiazepines, compound 7 thus shows several anomalous binding properties: HSA and the CNS receptor demonstrated opposite enantioselective discrimination; HSA has reversed enantioselectivity for compound 7; and HSA stereospecifically binds the low-affinity enantiomer. PMID- 9329180 TI - Excitatory amino acid receptor antagonists: resolution, absolute stereochemistry, and pharmacology of (S)- and (R)-2-amino-2-(5-tert-butyl-3-hydroxyisoxazol-4 yl)acetic acid (ATAA). AB - We have previously shown that (RS)-2-amino-2-(5-tert-butyl-3-hydroxyisoxazol-4 yl)acetic acid (ATAA) is an antagonist at N-methyl-D-aspartic acid (NMDA) and (RS)-2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid (AMPA) receptors. We have now resolved ATAA via diastereomeric salt formation using N-BOC protected ATAA and (R)- and (S)-phenylethylamine. Enantiomeric purities (ee > 98%) of (R)- and (S)-ATAA were determined using the Crownpak CR(-) and CR(+) columns, respectively. The absolute configuration of (R)-ATAA was established by an X-ray crystallographic analysis of the (R)-phenylethylamine salt of N-BOC-(R)-ATAA. Like ATAA, neither (R)- nor (S)-ATAA significantly affected (IC50 > 100 microM) the receptor binding of tritiated AMPA, kainic acid, or (RS)-3-(2 carboxypiperazin-4-yl)propyl-1-phosphonic acid, the latter being a competitive NMDA antagonist. Electrophysiological experiments, using the rat cortical wedge preparation, showed the NMDA antagonist effect as well as the AMPA antagonist effect of ATAA to reside exclusively in the (R)-enantiomer (Ki = 75 +/- 5 microM and 57 +/- 1 microM, respectively). Neither (R)- nor (S)-ATAA significantly reduced kainic acid-induced excitation (Ki > 1,000 microM). PMID- 9329181 TI - Exciton chirality of bilirubin homologs. AB - Bilirubin, the yellow pigment of jaundice, is a bichromophoric tetrapyrrole that readily adopts either of two enantiomeric, folded conformations shaped like ridge tiles and stabilized by a network of six intramolecular hydrogen bonds. Interconversion of these M and P helical chirality conformational enantiomers is rapid at room temperature but may be displaced toward either enantiomer by intramolecular non-bonded steric interactions. Introduction of a methyl group at the beta and beta' carbons of the propionic acid chains on the symmetric bilirubin analog, mesobilirubin-XIII alpha, shifts the conformational equilibrium toward the M or the P-chirality intramolecularly hydrogen-bonded conformer, depending only on the S or R stereochemistry at beta and beta', resulting in pigments with intense exciton coupling circular dichroism (CD) for the approximately 430 nm transition(s). Optically active synthetic analogs of bilirubin with propionic acid groups lengthened systematically to heptanoic acid (1-5) were synthesized and examined by spectroscopy to explore the influence of alkanoic acid chain length on conformation and intramolecular hydrogen bonding. In these diacids and their dimethyl esters (6-10), strong exciton chirality CD spectra are observed, and the data are correlated with molecular helicity. PMID- 9329182 TI - Absolute configurational assignment of self-organizing asymmetric tripodal ligand metal complexes. AB - The solution configuration of labile coordination complexes may be difficult to determine, even in cases in which the solid state structure is known. We have previously synthesized a series of chiral ligands which form pseudo-C3-symmetric complexes with ZnII and CuII salts that possess an available electrophilic coordination site. Molecular modeling of ZnII complexes of the chiral ligand N,N bis[(2-quinolyl)methyl]-1-(2-pyridyl)ethanamine (alpha-MeBQPA) showed that the spatial arrangement of the heterocyclic arms is controlled by a substituent on one methylene arm, resulting in the adoption of an enantiomeric conformation displaying a propeller-like asymmetry. In this paper we report the application of the exciton chirality method to the determination of the conformation of asymmetric metal-ligand complexes in solution. There is a good correlation between the predicted and the observed Cotton effects, demonstrating that the geometry in solution closely resembles that predicted by computational simulations and those obtained by X-ray crystallographic studies of metal complexes with racemic and enantiomerically pure ligands. The X-ray crystallographic structure of the first optically pure complex in this series is reported. PMID- 9329201 TI - Histophysiological observations on the external auditory meatus, middle, and inner ear of the Weddell seal (Leptonychotes weddelli). AB - The external auditory meatus, middle, and inner ear of the deep-diving Weddell seal (Leptonychotes weddelli) were studied with light microscopic, histological, and histochemical techniques in order to contribute to the open discussion on the orientation of this seal in the darkness of the deep Antarctic seas. The external auditory meatus is characterized by a well-developed venous plexus, single apocrine ceruminous, and numerous holocrine sebaceous glands and an incomplete tube of elastic cartilage. The tympanic membrane is comprised of two layers of radially and concentrically arranged collagen fibers and by elastic fibers which are concentrated in the outer part of the ear drum. The tympanic cavity is lined by a pseudostratified prismatic ciliated epithelium with goblet cells; a plexus of wide venous vessels marks the subepithelial lamina propria. The cochlea is about 10 mm high and forms about two and a half turns. The richly pigmented stria vascularis is well vascularized, while the cell-rich prominentia spiralis contains only single small blood vessels. The organ of Corti contains one row of inner and three rows of outer hair cells. Cells of Hensen, Claudius, and Boettcher are present. The basilar membrane is of comparatively uniform simple structure and is composed of abundant glycoproteins, proteoglycans, collagenous fibers, and the loose tissue of the tympanal layer. The spiral ligament is built up by abundant proteoglycans and a complex system of radial and concentric collagen fibers; close to the osseous wall of the bony cochlea it contains fine elastic fibers. The inner zone of the osseous wall of the cochlea strikingly contains hyaline cartilage. The thin lamina spiralis ossea is covered by a limbus spiralis with interdental cells secreting the lamina tectoria, which has a fibrous texture and contains glycoproteins and negatively charged components. PMID- 9329202 TI - The arterial system of the sperm whale (Physeter macrocephalus). AB - The angioarchitecture of the sperm whale is basically similar to that of other mammals, but it has specific attributes associated with the aquatic environment of this animal and its tolerance for deep and long diving. Specialized features include an expansive aortic arch, unusually far anterior localization of the arch, symmetrical branching of common carotid and subclavian arteries from the aorta, the absence of direct connection between internal carotid arteries and brain arteries, the absence of a costocervical artery, and the presence of a well developed occipital artery. The sperm whale has extraordinarily well-developed retia mirabilia, distributed in the cranial cavity, vertebral canal, neck and thoracic cavity, around the optic nerve, and in the walls of the uterus. These retia are more extensively developed in the sperm whale than in any other cetacean previously studied. PMID- 9329203 TI - Knowledge of pragmatic conversational structure. AB - Knowledge of conversational pragmatic structure was examined by asking 53 female volunteers to rate the naturalness of three versions of an appointment-making conversation from a beauty salon. One version was the naturally occurring conversation. The other two were its two most frequent reconstructions created by a separate group of subjects asked to put the scrambled natural conversation "back together again." A chi-square test and standardized deviates showed that the naturally occurring conversation was rated as the most natural one. This result is attributed to subjects' implicit knowledge about conversational pragmatic structure. The role that this knowledge might play in language comprehension is discussed. PMID- 9329204 TI - Does drawing attention to the referent constrain the way in which children construct verbal messages? AB - Tasks which assess children's speaker skills in the referential communication paradigm frequently mark the target item in some way to indicate which picture should be described. It was hypothesized that this procedure would interfere with effective scanning of the visual array (comparison processing) assumed to be necessary for the production of accurate messages. It was also predicted that, when one item was highlighted, more redundant features would be included in messages. An experiment which compared a marked with an unmarked target condition across three age ranges (6, 8, 10 years) found that message accuracy was not affected by highlighting. But significantly more redundant features were reported in the marked condition, indicating that, where redundancy is the focus of interest, a highlighting procedure should be avoided. Two age effects were also found: Message accuracy improved with age, while redundancy reduced with age. PMID- 9329205 TI - Temporal relationships between gaze and vocal behavior in prelinguistic and linguistic communication. AB - This work reports longitudinal evaluation of the temporal relationships between gaze and vocal behavior addressed to interactive partners (mother or experimenter) in a free-play situation. Thirteen children were observed at the ages of 1;0 and 1;8 during laboratory sessions, and video recordings of free-play interactions with mother and a female experimenter were coded separately for children's vocal behavior (vocalizations and words) and gaze toward their interactive partners. The difference between the observed and expected co occurrence of these two communicative behaviors was evaluated by transformation into z-scores. The most important findings are related to differences in the temporal relationship observed at age 1;0 between gaze and vocalizations and at age 1;8 between gaze and words. At the earlier age, the infants who exhibited greater coordination between gaze and vocal behavior than was expected by chance (z-score > +1.96) preferred to look at the interlocutor at the beginning of the vocal turn. Instead, when they were older and began to produce words, they frequently looked at the interlocutor at the end of the vocal turn. These results are interpreted as referring to characteristics of conversational competence in the prelinguistic and lingustic periods. Moreover, looking at the interlocutor at the beginning of the vocal turn at age 1;0 was found to be related to language production at age 1;8, highlighting a significant relationship between conversational competence during the prelinguistic period and language acquisition. PMID- 9329206 TI - Tacit integration and referential structure in the language comprehension of aphasics and normals. AB - Aphasics, brain-damaged patients with no language deficit, neurologically intact elderly subjects, and university undergraduates matched pictures to sentences having compelling tacit implications (e.g., the sentence The fox grabs the hen strongly invites one to assume that the fox will eat the hen). All groups made, for the same sentences, qualitatively similar referential errors consisting in choosing a tacit implication picture. Two auxiliary experiments using the same target sentences in other interpretive situations permitted ruling out the possibility that these errors were due to the putative intrinsic semantic properties of the sentences, showing that the sentences which were most liable to elicit integrative error varied from task to task. These results are interpreted within the conceptual framework which posits that reliable directions for interpretation are couched by the speaker in the very structure of his utterances (the utterance's referential structure) providing the hearer with means to restructure the relevant personal knowledge integrated into the interpretive process in accordance with the speaker's communicative intent. The determination of the referential structure (RSD) of utterances thus seems critical to their correct or, more precisely, conventional interpretation, and, along with the tacit integration of relevant sources of personal knowledge, constitutes the principal cognitive device enabling us to understand each other. But this device appears to be easily corruptible. It is suggested that many errors made by aphasics in language interpretation are due to a failure to follow all referential instructions, but that qualitatively similar failures also occur in normal subjects, though to a lessen degree. Language interpretation is a fallible process and aphasic errors provide remarkable clues for the understanding of its subtle referential mechanisms. PMID- 9329207 TI - Effects of repeated administration of dithiol chelating agent--sodium 2,3 dimercapto-1-propanesulphonate (DMPS)--on biochemical and haematological parameters in rabbits. AB - The effects of weekly intravenously administered a dithiol chelating agent-sodium 2,3-dimercaptopropane-sulphonate (DMPS)-in a single dose of 50 mg/kg/week for 10 weeks on biochemical and haematological parameters were studied in rabbits. DMPS was well tolerated, an increase in body weight was similar in the DMPS-treated and control animals. DMPS caused significant decrease in plasma calcium and vitamin E concentrations at the end of the experiment. No significant differences in haematological parameters between the DMPS and control groups were observed. A significant decrease in magnesium content in myocardial tissue was observed in the DMPS-treated rabbits. The above-mentioned biochemical changes should be taken into account in studies of possible chelating and radical scavenging effects of DMPS in various pathological conditions. PMID- 9329209 TI - The chronic anal fissure treatment by internal partial lateral sphincterotomy and following anal manometry. AB - Anal fissure remains one of the most common proctologic problems. A number of reports have advocated the use of partial internal sphincterotomy as a treatment of chronic anal fissure. Our study shows the results of a retrospective analysis of our patients who underwent lateral internal sphincterotomy for the treatment of chronic anal fissure. To determine long time results we examined random sample of 75 operated patients. Apart from taking careful history and patient's assessment of the operation effect, the patients were investigated "per rectum", and even possibly rectoscopically. To asscertain suitability and also security of properly done internal sphincterotomy, we performed anorectal manometric examinations in 53 patients controlled. To conclude our results, we can state that in 9 (12%) patients controlled some subjective complaints or certain pathologic findings connected with anal fissure or sphincterotomy were found. None of the people from the set of ours suffered from any complaints, which can be taken as stool incontinency. In no patients any change in pressure measured by anorectal manometry indicating incontinency was proved. PMID- 9329208 TI - Effect of alprazolam and ketamine on seizures induced by two different convulsants. AB - Effect of two anticonvulsants with different mechanism of action, i.e. alprazolam and ketamine, was tested in two types of seizure activity. The first one was induced by N-(3,5-dimethoxy-4-propoxyphenylethyl)-aziridine, the second one by pentylenetetrazol. While alprazolam alleviated both the minimal as well as major paroxysms, ketamine suppressed only major seizures. These effects are discussed in terms of the both N-methyl-D-aspartate and GABA receptors involvement. PMID- 9329211 TI - Hemothorax--a complication of subclavian vein cannulation. AB - Massive bleeding into pleural cavity after subclavian vein cannulation is a rather rare but very serious complication. Usually laceration of the venous wall is the cause. In patients where conservative treatment, i.e. pleural drainage, maintaining the circulatory volume, treatment of possible coagulopathy, etc. is ineffective, surgery has to be performed. Bleeding can be surgically managed either from posterolateral thoracotomy or direct subclavian vessel revision is possible after partial resection of the clavicle. Brachiocephalic vein bleeding can be approached and managed through median sternotomy. We present a case report of 22-year old man with hemothorax after subclavian vein cannulation. In our patient only complex surgical procedure enabled proper management of bleeding complication. PMID- 9329210 TI - Screening for organic acid disorders. AB - The detection of organic acidurias is a part of our screening programme for inherited metabolic diseases. Adapted procedure is differentiated and involves several steps: 1) thin-layer chromatography (TLC) in the case of an abnormal finding followed by 2) gas chromatography (GC). The next step of the investigation, using 3) gas chromatography mass-spectrometry (GS-MS) is reserved for more complicated and dubious analyses. In acutely sick patients and in the case of discrepancies between TLC results on the one hand, and clinical symptoms, supported by other laboratory findings on the other, the GC or GC-MS-analysis is performed immediately. Some examples of metabolic disorders, identified by this procedure, are presented. PMID- 9329212 TI - The history of otorhinolaryngology at the Charles University Faculty of Medicine in Hradec Kralove. PMID- 9329213 TI - Immunomodulatory functions and molecular regulation of IL-12. PMID- 9329214 TI - Structural and functional aspects of the IL-12 receptor complex. PMID- 9329215 TI - Early events controlling T-helper cell differentiation: the role of the IL-12 receptor. PMID- 9329217 TI - The immunostimulatory function of IL-12 in T-helper cell development and its regulation by TGF-beta, IFN-gamma and IL-4. PMID- 9329216 TI - Regulation of IL-12 receptor expression in early T-helper responses implies two phases of Th1 differentiation: capacitance and development. PMID- 9329218 TI - The role of IL-12 and IL-4 in Leishmania major infection. PMID- 9329219 TI - Initiation of T-helper cell immunity to Candida albicans by IL-12: the role of neutrophils. AB - The Th1/Th2 paradigm of acquired immunity is proving essential for a better understanding of immunoregulation in candidiasis and perhaps other fungal infections, conditions that may be life-threatening in humans and difficult to control by chemotherapy alone, especially in neutropenic and severely immunocompromised patients. In its basic conception applied to Candida infection in mice, this paradigm calls for: (i) an association between Th1 responses and the onset/maintenance of phagocyte-dependent immunity, critical for opposing infectivity of the commensal, focusing an infection, and clearing the yeast from infected tissue; (ii) the ability of the yeast to activate Th2 response as an evasive strategy; (iii) the reciprocal regulation of Th1 and Th2 responses, resulting in a dynamic balance between these two types of reactivity. This balance, in concert with a variety of environmental factors, may regulate the status of the yeast as a commensal or pathogen in the mucosal tissue of colonized humans, but may also determine the outcome of deep-seated systemic infections once hematogenous dissemination of the yeast has occurred. An important corollary of this hypothesis may be the possible combined effects on Th immunity of Candida carriage/infection and various disease states. While immune deficiency or dysregulation, resulting in an altered cytokine balance as may occur in AIDS, can reasonably be expected to increase local infectivity of the yeast, it is even more intriguing that antifungal chemotherapy will resolve some of the unusual skin (atopic dermatitis-like) disorders frequently observed in this clinical setting, patients with AIDS. Besides, an immunopathologic role for Candida has been suggested for atopic dermatitis, atopy, and other conditions, overtly associated or not with Candida. Thus, the Th cell dichotomy to Candida may have important implications not only for regulation of the balance between commensalism and infection, but may also contribute to the onset or dominance of Th2 responses in other disease states. A similar example, although with different effects, may be provided by the temporary improvement seen in atopic dermatitis patients in concomitance with acute severe infections, an effect that has been proposed to result from transient down-regulation of the predominant Th2 cell reactivity. With a view to either controlling Candida infections or opposing Candida-related immunopathology, the promotion of yeast-specific Th1 responses appears to be a promising immunotherapeutic approach. This, in principle, could be achieved by subtraction of Th2 cytokines or by administration of Th1-promoting cytokines. However, our initial studies with exogenous IL-12 were unsuccessful, suggesting that the recombinant cytokine: (i) is unable to oppose Th2 differentiation driven in vivo by IL-4/IL-10; (ii) may induce endogenous IL-10 production as a regulatory response, and (iii) may potentate local inflammatory responses in gastrointestinal infection or even trigger IFN-gamma-dependent mechanisms of fungal septic shock.. More recent studies seem to provide encouraging results, at least under specific conditions of testing. In acute candidemia, neutrophils appear to be a major source of the directive cytokines, IL-12 and IL-10, thus contributing to the selection of Th1 and Th2 cell responses to LVS or virulent infection, respectively. Neutrophils may also be an important source of IL-10 released in response to challenge with exogenous IL-12. As a result, the Th1-promoting role of IL-12 may be largely unopposed (by IL-10 induction) in neutropenic mice, which would otherwise succumb to LVS challenge. These animals are, in fact, cured by replacement therapy with IL-12 and acquire durable, Th1-associated anticandidal protection. These findings may be very important for immunotherapy of fungal infections in humans. Neutropenic patients are those at the highest risk for developing systemic candidal infections. (ABSTRACT TRUNCATED) PMID- 9329220 TI - Identification and characterization of protozoan products that trigger the synthesis of IL-12 by inflammatory macrophages. PMID- 9329221 TI - Antitumor activities of IL-12 and mechanisms of action. PMID- 9329222 TI - Targeting IL-12, the key cytokine driving Th1-mediated autoimmune diseases. PMID- 9329223 TI - Notes from a clinical information system program manager. A solid vision makes all the difference. AB - Today's CIS manager will create a vision that connects computerization in ambulatory, home and community-based care with increased responsibility for patients to assume self-care. Patients will be faced with a glut of information and they will need nursing help in determining the validity of information. The new vision in this environment will focus on integration, interoperability, and a new definition for patient-centered information. Creating a well-articulated vision is the first skill in the repertoire of a CIS manager's tool set. A vision provides the firm structure upon which the entire project can be built, and provides for links to life-cycle planning. This first step in project planning begins to bring order to the chaos of dynamic demands in clinical computing. PMID- 9329224 TI - Using a multimedia slide show with interactive television. PMID- 9329225 TI - The application of pen-based computer technology to home health care. AB - The purpose of this project was to study the applicability of pen-based computer technology to home health care through the development of a pen-based computer system for a Hospital/Community-Patient Review Instrument (H/C-PRI) used for nursing home placement. The sample included nurses (n = 12) from the four regional Visiting Nurse Service of New York offices, as well as all patients on whom a H/C-PRI was completed during the pre-period (n = 238) and patients on whom a H/C-PRI was completed during the post-period (n = 238). The quality of documentation was higher for patients whose H/C-PRI was performed using the pen based computer (0% calculation errors) than for those patients whose H/C-PRI was documented in the usual manner (11% calculation errors). PMID- 9329226 TI - Ergonomic nursing workstation design to prevent cumulative trauma disorders. AB - The introduction of computerized nursing information systems offers health care institutions an opportunity to take a new look at safety issues related to nursing workstation design. Industrial studies have investigated the injuries sustained by clerical workers who spend long periods of time at their computers. Cumulative trauma disorders (CTDs) are the most common injuries associated with computerized workstation use. They account for nearly 90,000 injuries each year in the United States. Typical CTDs include back pain, strain of the neck, shoulders and eyes, and carpal tunnel syndrome. As the information handling work of nurses is increasingly computerized, the incidence of computer-related injury is expected to increase. Injury rates can be reduced by ergonomic workstation design. An assessment of potential risks associated with the equipment installed should be done as part of workstation design. Risk identification is a prerequisite for avoiding injuries by designing workstations that protect human health. The ergonomic principles learned and tested on office workers are addressed and extrapolated to nursing workstation design. Specific suggestions for design of sitting and standing workstations are presented. PMID- 9329227 TI - Computerized NCLEX-RN and NCLEX-PN preparation programs. Comparative review, 1997. AB - The computerized test taking market continues to expand to meet the needs of nursing students studying to prepare for the NCLEX-RN and the NCLEX-PN. In addition, currently registered nurses may choose to use these software programs to review clinical knowledge in areas in which they are not currently practicing. This article reviewed software designed for personal use, review books with complete disks, on-site institutional testing and consultation, and school of nursing LANs. The costs of software for personal use is priced reasonably and provides flexibility for students to use as their schedule permits. The cost of institutional licenses is moderate and most programs provide multiple on-site use rights. The marketplace has responded to the computerized NCLEX testing now in place nationally. As new formats are developed and new uses identified, nursing faculty and students can expect to see an expanded use of computerized testing. PMID- 9329228 TI - Using the Internet for data collection in nursing research. AB - This article examines how the internet may be used as a tool for data collection in nursing research. An overview of the demographic composition of the Internet population is outlined and discussed as a constraint on the type of research that can be undertaken using the Internet. Methods of data collection such as e-mail and WWW questionnaires are discussed as well as the possibility of virtual focus groups. Some of the difficulties and advantages that may confront the researcher wishing to undertake research using the Internet are outlined. PMID- 9329229 TI - Wolff Award 1997. Involvement of a Ca2+ channel gene in familial hemiplegic migraine and migraine with and without aura. Dutch Migraine Genetics Research Group. AB - A gene for familial hemiplegic migraine, a subtype of migraine with aura, was assigned to chromosome 19p13. In this region, we identified a brain-specific P/Q type calcium-channel alpha 1A-subunit gene, CACNA 1A, with 47 exons covering 300 kb. Sequencing of all exons and their flanking surroundings revealed polymorphic variations, including a (CA)n-repeat and a (CAG)n-repeat in the 3' untranslated region. In patients with familial hemiplegic migraine, we found four different missense mutations in conserved functional domains. One of the mutations has occurred on two different haplotypes in unrelated familial hemiplegic migraine families. Moreover, in episodic ataxia type 2, we found two mutations disrupting the reading frame. Thus, familial hemiplegic migraine and episodic ataxia type 2 can be considered as allelic channelopathies. Involvement of this familial hemiplegic migraine locus in migraine with and without aura was demonstrated by sib-pair analysis. We showed an increase of shared marker alleles of locus D19S394, which is tightly linked to the gene. The association between the alpha 1A calcium channel and familial hemiplegic migraine, and the increase of shared alleles in migraine-affected sib-pairs, have uncovered a new pathway for the pathophysiology of migraine. This finding may provide a rationale for the development of specific prophylactic therapy for migraine and other (paroxysmal) cerebral disorders. PMID- 9329230 TI - Alteration of central excitation circuits in chronic headache and analgesic misuse. AB - Central excitatory circuits could be involved in the pathophysiology of pain; particularly, the genesis of chronic pain. The "second pain" is the sensation that follows the initial pain after an appropriate nociceptive stimulus. The second pain is amplified by repeating the stimulus after brief intervals (temporal summation). This phenomenon is the psychophysical correlate of the excitatory pain circuits. The temporal summation of the second pain was evaluated in four groups of subjects; one group affected by migraine without aura, one by episodic tension headache, one by chronic daily headache, and a group of healthy subjects. A percutaneous electrical shock was used as the nociceptive stimulus. The intensity of the second pain was significantly greater in the group of patients with chronic headache in comparison with the other groups. The patients with chronic headache were subdivided into three groups on the basis of their clinical history: a group with transformed migraine; a group with chronic headache ab initio, a form related to the first one; (both groups suffered from chronic daily headache with a frequent superimposition of episodes of migraine attacks) and the third group consisted of patients with chronic tension headache. The temporal summation of the second pain was altered in the first two groups. The patients with chronic migraine abused ergotamine given as a symptomatic drug. Those who were able to discontinue this drug were retested and reported a decrease of the second pain in comparison to the previous measurements. The results of the present study indicate that central excitatory circuits could be involved in the mechanism leading to the development of chronic daily headache. PMID- 9329231 TI - Photophobia and phonophobia in migraineurs between attacks. AB - This study investigated whether migraineurs are more sensitive to light and sound while headache-free than are healthy people. Fifty-two migraineurs (mean age 39 years) were selected using the International Headache Society diagnostic criteria for migraine. Forty-eight healthy controls were matched for age, sex, and race (mean age 36 years). Visual and auditory discomfort thresholds were measured by exposing subjects to increasing light and sound until they complained of discomfort. There were significant differences between groups in both the light discomfort threshold (P < 0.00005) and the hearing discomfort threshold (P < 0.0005). The thresholds for both were lower in the migraineurs. Overall, for both groups together, there was a significant negative correlation between light discomfort threshold and age (correlation coefficient -0.2276, P = 0.011), but not for the hearing discomfort threshold and age (P = 0.275). The results show that the migraineurs were significantly more sensitive to light and sound when headache-free than were healthy controls. The apparent increased intolerance to light in both groups together noted with increased age, did not apply to the migraine group. PMID- 9329232 TI - The enhanced ciliospinal reflex in asymptomatic patients with cluster headache is due to preganglionic sympathetic mechanisms. AB - An amplified ciliospinal reflex response has been documented in patients with cluster headache, lacking a Horner-like syndrome. The mechanism is unknown. Tentatively, it may be due to an increased release of monoamines from post ganglionic sympathetic nerve endings or an increased density of postsynaptic adrenergic receptors in the dilatator muscle of the iris. The instillation of a 1% phenylephrine solution into the conjunctival sac induces mydriasis by stimulating postsynaptic adrenergic receptors in the dilatator muscle of the iris, while the instillation of a 2% tyramine solution causes mydriasis by releasing noradrenaline from the presynaptic sympathetic nerve terminals in the iris. According to these premises, a positive correlation should be expected between the ciliospinal reflex response and the pupillary response to tyramine, if the enhanced ciliospinal reflex response was due to an increased presynaptic release of monoamines. No such correlation was found. Nor was there any positive correlation between the ciliospinal reflex response and the pupillary response to phenylephrine, contradicting an increased density of postsynaptic monoaminergic receptors in the dilatator muscle of the iris as the explanation. However, there was a significant positive correlation between the pupillary responses to phenylephrine and tyramine, ruling out any functionally caused "denervation" hypersensitivity in the dilatator muscle of the iris. It is concluded that the amplified ciliospinal reflex response in cluster headache patients (lacking a Horner-like syndrome) reflects compensatory pathophysiological mechanisms proximal to the third-order sympathetic neuron. PMID- 9329233 TI - Mental stress of long duration: EMG activity, perceived tension, fatigue, and pain development in pain-free subjects. AB - The study examined the relationship between pain development in the shoulder, neck, and facial regions and the EMG activity of underlying muscles, during prolonged exposure to a mental stressor. The subjective perception of tension and fatigue was recorded. Thirty-six subjects were exposed to a two-choice reaction time test for 1 hour. Electromyographic (EMG) recordings were performed bilaterally over the frontalis, temporalis, splenius, and trapezius muscles. Pain and perceived tension were scored on a visual analog scale, and fatigue on a Borg scale. Pain development was most pronounced in the shoulder and neck region. There was a weak tendency of those reporting pain in the shoulder region to generate higher EMG activity in the trapezius relative to those with no shoulder pain at the end of the test. No such relationship was observed for the other muscles. Perceived tension during the test was weakly related to pain and strongly related to fatigue at the end of the test, but not to EMG level. It is concluded that the mean level of the EMG response is of little consequence for pain development during stressful conditions. It is argued that other physiological responses such as prolonged activity in low-threshold motor units, whereby the surface EMG response can serve as a marker, can be important for shoulder pain originating in the trapezius muscle. PMID- 9329234 TI - Migrainous syndrome with CSF pleocytosis. SPECT findings. AB - Brain single photon emission computed tomography (SPECT) findings are described in four adult patients with the transient syndrome of headache with neurological deficits and cerebrospinal fluid (CSF) pleocytosis. Focal deficits consisted of right-sided hemisensory changes with or without motor dysphasia or dysarthric speech (n = 4) and confusional episodes (n = 1). All patients had a CSF pleocytosis (with a mean of 309 cells/mm3 on the first spinal tap; range 75 to 590) and an elevated total protein (mean 130.5 mg/dL; range 70 to 193). The EEG showed excessive focal slowing (n = 2). A technetium Tc 99m hexamethyl propylenamine oxime (HMPAO) brain SPECT was performed during a symptom-free period, within 8 and 25 days after the onset of symptoms (n = 4). Three patients showed a decreased tracer uptake in the anterior left hemisphere, topographically consistent with the neurological deficits and EEG slowing. One patient showed no abnormalities. These findings indicate either focally impaired neuronal metabolism or hypoperfusion in regional cerebral blood flow, which could bear some relationship with the clinical features. The possibility that SPECT abnormalities may represent an epiphenomenon was also considered. PMID- 9329235 TI - Cerebral vasomotor changes in the transient syndrome of headache with neurologic deficits and CSF lymphocytosis (HaNDL). AB - We report two patients with the recently described transient syndrome of headache with neurologic deficits and CSF lymphocytosis (HaNDL). Transcranial Doppler sonography performed during and after attacks of HaNDL showed asymmetrical decreases or increases in blood flow velocity of the middle cerebral artery, accompanied by increases or decreases in pulsatility suggesting fluctuations of arteriolar tone. The findings demonstrate focal vasomotor disturbances that link the transient headaches and deficits of HaNDL with attacks of migraine. PMID- 9329236 TI - Serotonin syndrome presenting with migrainelike stroke. AB - Serotonin syndrome is the result of the drug interaction that enhances serotonergic tone in the central nervous system. The neurological manifestations are transient in most of the reported cases with rare fatality. CASE DESCRIPTION: A 30-year-old woman developed ischemic infarction of the right temporoparietal and cerebellar hemispheres 48 hours after administration of a large dose of clomipramine. This drug was given within 24 hours of discontinuation of high-dose fluoxetine, both of which are known to enhance serotonergic tone in the central nervous system. The distribution of the ischemic infarction in this case did not respect the major territorial arteries and was similar to spreading oligemia/ischemia that is described in migraine. A similar pathogenesis may exist for stroke in serotonin syndrome and in migraine. PMID- 9329237 TI - Painful trigeminal neuropathy: clinical and pharmacological observations. AB - A 74-year-old woman had a 5-year history of constant burning pain and numbness of the central face of subacute onset. The central region of the face, oral cavity, and nose lacked all sensation. Corneal reflexes and the jaw jerk were absent. Blood tests, rectal biopsy, neurodiagnostic studies, and surgical exploration of the trigeminal nerve were normal. Blink reflexes were absent. Facial nerve motor latencies and EMG of the facial and masseter muscles were normal. Responses to the thermoregulatory sweat test, intradermal histamine, and simulated diving were present. Oral administration of 500 mg L-dopa aggravated her pain and produced transient hypalgesia in the C2 through C6 dermatomes. Infraorbital nerve biopsy demonstrated loss of large myelinated fibers. IN CONCLUSION: (1) Only the central region of the face is exclusively supplied by the trigeminal nerves. (2) Somato autonomic reflexes coupled with electrophysiological studies localized the lesion to the large fibers. (3) Large fiber loss and central brain stem reorganization may explain the burning pain. (4) Dopamine may modulate trigeminal nociception. PMID- 9329238 TI - A late 'migraine': the only symptom of an intrasellar aneurysm. AB - The history of a 62-year-old woman affected by an intrasellar aneurysm is described. A migrainelike headache was, for many years, her only complaint. PMID- 9329239 TI - Migraines and tannins--any relationship? PMID- 9329269 TI - Uses of dietary reference intakes. AB - The correct reference value must be used for its intended purpose, which usually involves either planning for an adequate intake or the assessment of adequacy of intake. It is anticipated that future publications will address the interpretation and uses of DRIs in more detail in order to assist both the health professional and those interested in nutrition policy and analysis. PMID- 9329268 TI - Dietary reference intakes. AB - The term Dietary Reference Intakes (DRIs) is new to the field of nutrition. It refers to a set of at least four nutrient-based reference values that can be used for planning and assessing diets and for many other purposes. The development of DRIs replaces the periodic revisions of Recommended Dietary Allowances (RDAs), which have been published since 1941 by the National Academy of Sciences. This is a comprehensive effort being undertaken by the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes of the Food and Nutrition Board, Institute of Medicine, National Academy of Sciences, with the involvement of Health Canada. PMID- 9329270 TI - Origin and framework of the development of dietary reference intakes. AB - This report is the first in a series of publications resulting from the comprehensive effort being undertaken by the Food and Nutrition Board's Committee on the Scientific Evaluation of Dietary Reference Intakes and its panels and subcommittees. The history and rationale behind the development of the Dietary Reference Intakes are discussed. PMID- 9329271 TI - Calcium and related nutrients: overview and methods. PMID- 9329272 TI - A model for the development of tolerable upper intake levels. PMID- 9329273 TI - Aesthetic inquiry and the art of nursing. AB - This article describes the development of aesthetic inquiry and the emerging conceptualization of the art of nursing as an art form. Aesthetic knowing, which emerges from aesthetic inquiry, is described as connoisseurship of the art of nursing and includes appreciation of the art form and insight into meanings of the art. A method of aesthetic criticism is described that links artistic experience, history, form, alternate meanings, and future possibilities. The inquiry yielded two essential elements of the art of nursing, movement and narrative, which, when manifested as an art form have the capacity to shift experience into a different realm. PMID- 9329274 TI - Social dancing in the care of persons with dementia in a nursing home setting: a phenomenological study. AB - The purpose of this study was to describe the phenomenon of social dancing in the care of persons with dementia in a nursing home setting. Social dancing is an activity that has taken place once a month regularly during the last 10 years at a nursing home in Stockholm. The period of data collection for this study was the year 1995. At the time of the investigation, the subjects were in special units for persons with dementia. The analysis is based on the data contained in five 45 minute video tapes. All videotapes were analysed based on Husserl's philosophy and Giorgi's method of phenomenological analysis. The results suggested that dance music was a good stimulus for making social contacts. The earlier-trained social patterns, old social habits, and general rules seemed to awaken to life in the persons with dementia. It was important that the caregivers showed individual creativity, spontaneity, and supportive nursing care. Social dancing at the nursing home was found in this study to be very positive and successful for patients with dementia. PMID- 9329275 TI - The impact of HIV on emotional distress of infected women: cognitive appraisal and coping as mediators. AB - This study examined the role of psychological factors as mediators of the impact of HIV-related stressors on emotional distress of a clinic-based sample of 264 HIV+ women. Based upon Lazarus and colleagues' cognitively oriented theory of stress and coping, causal modeling was used to test for mediating effects of cognitive appraisal (intrusive thoughts and perceived stigma) and coping variables (avoidance and fatalism) on emotional distress within the context of HIV-related stressors (functional impairment and work performance impairment). The findings supported the mediating effects of cognitive appraisal but not of the coping variables. Consistent with theory, the effect of HIV-related stressors on emotional distress was indirect through cognitive appraisal; however, there were no significant direct effects of HIV-related stressors, fatalism or avoidance on emotional distress. The causal model accounted for significant portions of variance in emotional distress (R2 = .49) and the model fit, as a whole, was more than adequate. The findings indicate that how HIV+ women think about HIV-related stressors is an important factor that may account for individual variability in the ability to maintain a sense of subjective well being in the face of a devastating fatal disease. PMID- 9329276 TI - High touch in high tech: the presence of relatives and friends during resuscitative efforts. AB - Hospital emergency departments traditionally have policies that require relatives and friends to wait out the resuscitative attempt of their loved ones in a counseling room. In recent years, this widespread practice has been questioned. In some hospitals, family members and friends are given the option to attend the resuscitative effort. Based on in depth interviews, three different resuscitation perspectives to which health care providers subscribe have been identified. According to the advocates of the survival framework, the only goal that a resuscitation should achieve is to save a human life. In the bifurcated perspectives, a resuscitation has two separate goals: saving lives and taking care of family members. Health care providers who subscribe to the holistic framework are still concerned with survival of the patient, but significant others become participants in the resuscitation process. PMID- 9329277 TI - Leprosy elimination campaigns--reaching every patient in every village. PMID- 9329278 TI - Outbreak of hand, foot and mouth disease in Sarawak. Cluster of deaths among infants and young children. PMID- 9329279 TI - Socio cultural factors in leprosy: implications for control programmes in the post leprosaria abolition years in Nigeria. AB - A questionnaire was administered to 53 male and female leprosy patients aged 17 78 years, randomly selected from four clinics in two Local Government Areas of the Eastern part of Nigeria to determine the impact of socio cultural factors on, and also to predict the chances of compliance at, leprosy control measures. About 60% of the patients indicated that traditional concepts were the likely factors explaining the aetiology of leprosy. Four patients were convinced about the microbial aetiology of leprosy. Traditional concepts of leprosy aetiology were significantly associated with mode of entry into the control programme (P < 0.025) and tendency to live within the vicinity of the leprosy clinics (P < 0.01) but not with clinic attendance rate of leprosy patients. The distance of patient's abode from clinic attended, some formal education and whether or not patient's spouses were alive, were not significantly associated with clinic attendance rate. There was a significant association between maleness, age less than 55 years (P < 0.025) as well as negative family attitude (P < 0.05) and irregular clinic attendance. Its implications therefore are to broadly categorize these at risk group at first contact and target towards them patient-holding methods as well as health education, targetted towards patients, their relations and the community. PMID- 9329280 TI - Pattern of out-patient drug treatment of hypertension in Korle-Bu Teaching Hospital, Accra. AB - In a retrospective study of the pattern of drugs used in the initial treatment of hypertension, 300 case notes of hypertensive patients attending medical clinics at the Korle-Bu Teaching Hospital, Accra and whose treatment were initiated during the period 1973-1993, were reviewed. The mean age of patients was 55 years, mean pre-treatment systolic and diastolic pressures were 179.5 +/- 25. 5 and 108.5 +/- 14.2 mm Hg, respectively, with 85% of patients being female. The frequencies of individual drugs prescribed for the initial treatment of hypertension were: diuretics (90%), reserpine (46%), methyldopa (31%), and propranolol (30%). Single drug treatment was prescribed for 18%, two drugs for 60% and three or more (multiple drugs) for 22% of patients. The mean number of drugs per patient was 2.2 Patients prescribed multiple drugs had pre-treatment systolic and diastolic blood pressures which were significantly (p < 0.01) higher than those prescribed 2 drugs which were, in turn, higher (p < 0.001) than those prescribed single drugs. The results showed that during the period 1973-1993, diuretic based "stepped care" therapy was the main first line anti-hypertensive drug management regime. "Old" anti-hypertensive drugs were more commonly used than newer ones. The cost and availability drugs and the pretreatment blood pressure were probably the main determinants of the choice of the type and number of drugs prescribed. PMID- 9329281 TI - Epilepsy: knowledge, attitude and practice in literate urban population, Accra, Ghana. AB - A cross sectional survey was conducted among Government workers and the general public in Accra, Ghana. A total of 380 persons were interviewed. Almost everybody could describe accurately, an epileptic person. However, 172 (45.3%) out of the 380 respondents did not know the cause of epilepsy, and 37.6% did not know how it could be treated. Out of the 358 responses to the cause of epilepsy, 114 (31.8%) said it was inherited disease, 100 (27.9%) said it was due to witchcraft/juju or spiritual. With respect to treatment, 150 out of 333 responses mentioned sending the individual to the medical doctor, 95 (28.5%) said the use of herbs/visits to fetish priest, 59 (17.7%) suggested prayers, 20 (6.0%) said to do nothing. For prevention, 77 (29.1%) out of 319 responses indicated prayers, 49 (15.45%) cautioned marrying into epileptic family, and 13 (4.1%) responses indicated not to touch patient fitting. Those who answered "don't know" regarding knowledge about epilepsy were mostly the young, the lower educational status and the single respondents. However, the most important characteristic of the respondent that was associated with the appropriateness of the responses was the educational status. Although a lot of misconceptions about epilepsy existed in the study population, e.g. epilepsy can be spread by contact and that epileptics must be isolated or avoided, several respondents would share a room, eat or employ persons with epilepsy. The study has shown that the traditional beliefs and attitudes about epilepsy are still held firmly by the adult working population and that the educational level of the respondent was positively related to the appropriateness of the responses. It is therefore suggested that additional efforts must be made to increase the knowledge of the general population through the use of social marketing strategies in order to improve the management of persons with epilepsy. PMID- 9329282 TI - Common psychiatric disorders among the elderly attending a general psychiatric out patient clinic in Accra, Ghana: a five year retrospective study (1989-1993). AB - Details of thirty-five (35) elderly patients aged from sixty (60) years and upwards, who had attended a general psychiatric outpatient clinic, and duly registered as new patients between 1989-1993 were analysed. Depressive illness using the ICD-10 diagnostic criteria was found to be the predominant diagnostic category. This was followed by Dementia and Paranoid delusional disorder. No significant psychosocial contributory factors were elicited among the depressives. However, among the demerits there was a strong association with a previous history of excessive alcohol intake, especially the locally brewed Gin Akpeteshie among the males. Paranoid delusional disorder was found to be commoner among females, a third of those having had similar disorder in the past, and was associated with partial blindness and partial deafness. The majority of the patients were living with relatives. The implications of these findings with regards to future planning and the care of the elderly are discussed. PMID- 9329283 TI - Experience in managing splenic trauma on the Jos Plateau. AB - We studied retrospectively, fifty eight (58) patients with splenic injury admitted to Jos University Teaching Hospital between October 1988 and September, 1995. Forty-nine were males while nine were females (M:F = 5.4:1). The age ranged from 5 to 50 years with a mean of 24.5 years. The highest incidence was recorded in the third decade of life. Road Traffic Accident (RTA) was the commonest (75.8%) cause of splenic injury; others were fall from heights 7 (12.1%), blows to the abdomen 5 (8.6%) and stab wounds to the abdomen 2 (3.5%). Of the 58 cases, 56 (96.5%) were blunt abdominal injuries while 2 (3.5%) were penetrating stab injuries. All had laparotomy. 13 (22.4%) sustained Upadhyaya and Simpson's type 1 injury, 18 (31.0%) type II, 12 (20.7%) type III and 15 (25.9%) type IV injuries. Of the 58 patients, 29 (50%) had total splenectomy without heterotopic autotransplantation (HAT); 21 (36.2%) had splenectomy with HAT, while 8 (13.8%) had splenorrhaphy with omentoplasty. The average number of units of blood transfused was 2.3 units per patient. There were four (6.9%) deaths--two as a result of shock and multiple organ failure and another two died as a result of pulmonary embolism. However the commonest post operative complications were chest and wound infections. The rate of splenic salvage in this study was low despite the fact that most of these patients sustained types I and II injuries. We believe a greater salvage rate could be achieved and the trend in our centre now is toward splenic conservation. PMID- 9329284 TI - Trends in prevalence and pattern of substance use among secondary school pupils in Ilorin, Nigeria. AB - We present trends data on the prevalence and pattern of substance use among secondary school pupils in Ilorin, derived from comparing the finding of two consecutive cross-sectional surveys. From a sample of six schools, 1041 and 848 pupils 1988 and 1993 respectively, completed anonymously a 117-item WHO self report substance-use questionnaire. The analyses cover responses on the current and lifetime use of eleven substances, their frequency of use, and the effect of gender and school location on use trends. Although a significant increase in current use rates was recorded for alcohol, cannabis, mild stimulants and hypnosedatives, all of these substances (except stimulants) showed a shift towards less frequent use in 1993. The only consistent gender effect was found for smoking, which remained significantly a male activity. The significant increases found in the current use of cocaine, organic solvents and hallucinogens are difficult to substantiate, thus prompting the suggestion for further corroborative qualitative studies. There was a trend towards greater involvement of the rural school's respondents in substance use in general. The implications of the findings with respect to policy issues on substance-use prevention programmes for youths in the Ilorin metropolis are discussed within the context of the limitations of the study design. PMID- 9329285 TI - Maternal mortality following post-partum haemorrhage in Calabar a 6-year review. AB - During the 6-year period (January 1987 to December 1992) a maternal mortality rate of 16.2 per 1,000 deliveries and a post-partum haemorrhage case fatality rate of 2.2 percent were recorded at the University of Calabar Teaching Hospital. Sixteen of the 62 unbooked patients with primary post-partum haemorrhage died giving a case fatality rate of 25.8% as compared with a case fatality rate of 0.8% for booked patients with primary post-partum haemorrhage. A case fatality of 5.4% from induced labour compared with 1.8% from spontaneous labour was recorded, while assisted vaginal delivery had a case fatality rate of 4.7%. Uterine atony, retained placenta and coagulation defect in that order were the main causes of maternal death from primary post-partum haemorrhage. The case fatality rate amongst primigravidas now seems to be the same as that of grandmultipararas traditionally classified as high risk. Lack of adequate supervision of junior residents formed the major defect in the management of patients in this study. PMID- 9329286 TI - Blood glucose responses to mixed Ghanaian diets in healthy adult males. AB - The blood glucose responses to five Ghanaian carbohydrate sources, unripe plantain, Ga kenkey, Gari, rice and yam, as part of mixed meals were determined in ten healthy young nondiabetic adult males aged 25.6 +/- 2.6 years with a BMI of 20.9 +/- 2.4 kg/m2. Ga kenkey showed the least changes in blood glucose responses as measured by the glycemic index. Yam exhibited the least favourable blood glucose responses. Significant difference were observed between the glycemic indices of kenkey and yam; Kenkey and gari (p < 0.01); rice and yam, plantain and yam (p < 0.05). Further studies of these carbohydrate sources are required in diabetics to ascertain their suitability as carbohydrate sources in Ghanaian diabetics. PMID- 9329287 TI - Determinants of tetanus toxoid immunization of parturient women: a community based study in Rivers State of Nigeria. AB - A community-based study carried out in the Rivers State of Nigeria on tetanus toxoid immunization status of parturient women showed a complete, partial and no coverage status of 41.2, 17.0 and 41.8 per cent respectively of women surveyed. Formal education to the secondary school level was very strongly associated with complete coverage status (p < 0.001). Also of importance was the peculiar geographical terrain of the state since the place of obstacles as a negative factor was significantly more pronounced among the riverine community (p < 0.001). Generally, communities in the state will require more logistics support than elsewhere in the country for any intervention measure to have an appreciable impact and on the long term the institution of measures aimed at raising the literacy level of the population as a whole will bring about overall improvement in the vaccine coverage in the state. PMID- 9329288 TI - Comparison of albendazole and levamisole chemotherapy on prevalence and intensity of common soil-transmitted helminth infections in school children, Sierra Leone. AB - A comparison of two studies performed in Sierra Leone on the effect of anthelmintic, chemotherapy, levamisole, albendazole or a placebo in children aged 6 to 10 years on the prevalence and intensity of common soil transmitted helminths (S-THs) infections is presented. In total 501 children were screened, and 394 successfully follow-up. At baseline their overall prevalence and intensity (epg) of Ascaris, Necator and Trichuris were 34% (2,877), 22% (284) and 39% (266) respectively. At baseline there was no significant difference in the intensity of S-THs infections in the different treatment groups but the prevalence of Necator was significantly higher in the levamisole than the albendazole group (p < 0.05). At follow-up both albendazole and levamisole significantly reduced prevalence and intensity of Ascaris and Trichuris. Only albendazole significantly reduced those of Necator. The placebo group had no significant change in prevalence but a significant increase in intensity of all S THs. PMID- 9329289 TI - The family planning aspects of the practice of traditional healers in Ibadan, Nigeria. AB - The family planning aspects of the practice of traditional healers in Ibadan, a large city in south west Nigeria, was investigated by means of a questionnaire survey of 193 traditional healers. The findings revealed that between 13% and 53% agreed with certain cultural beliefs which tend to increase fertility and that their perceptions of ideal child spacing is most commonly 2-3 years. Only 13% think a couple should have a specified number of children; a large proportion think the number should be "as God wills" (42%) or as many as the couple has resources to cope with (42%). Nearly all think that traditional healers and orthodox health professional should work together in the area of family planning. While most of them recommend traditional methods of contraception (such as beads and herbs) to their clients, up to 22% recommend modern family planning methods such as condoms and oral contraceptive pills. The implications of these findings for family planning programmes and the possibility of the involvement of traditional healers in the promotion of modern family planning methods are discussed. PMID- 9329290 TI - Intraocular pressure asymmetry-Topcon Computerised Tonometer CT-20. AB - The Topcon Computerised Tonometer CT-20, a non-contact tonometer (NCT), was used to measure intraocular pressure (IOP) in mmHg in 1,226 subjects above the age of 30 years with no ocular pathology. The mean IOP was 14.41 mmHg and the standard deviation 3.43. The mean IOP far was higher in female (14.60) than male (14.22). This gender difference in the mean IOP was significant; t stat = 2.69, P < 0.5. Right IOP exceeded left IOP and this was highly significant in both male (t stat 4.03) and female (t stat 5.64) p < 0.01. This IOP asymmetry approximates a normal distribution. The mean of the differences (right-left IOP) and standard deviation for all subjects were 0.57 and 2.26 respectively. The range for the differences was- 10 to 10 mmHg. Asymmetry was greater in females than males (t stat = 2.05, p < 0.05). Knowledge of these statistics will help in decisions concerning glaucoma screening with NCT. PMID- 9329291 TI - Superficial Salmonella abscesses in two siblings with sickle cell diseases. AB - Salmonella infection in sickle cell disease patients is generally well-known but presentation as superficial abscess is relatively uncommon. Two sisters aged 4 1/2 and 6 years presented with superficial subcutaneous abscesses that were caused by the same strain of Salmonella enteriditis group C. Despite in vitro susceptibility with a MIC of 0.03 microgram/ml and an adequate dosage of ciprofloxacin there was a relapse with widespread dissemination of the same organism in the younger sister who subsequently developed multiple osteolytic infections. Change of treatment to chloramphenicol produced a cure in both patients. PMID- 9329292 TI - What is so special about special education? AB - There is nothing special about special education. Educational methods have not changed significantly in at least 2,500 years. IQ tests were developed to identify those in need of special education, with the intention of developing appropriate educational methods. Effective special educational methods have yet to be developed. IQ tests are diagnostic but not prescriptive. Effective special educational methods will not be developed until (a) individual differences in student characteristics beyond IQ scores are recognized and understood and (b) educators focus on specific and realistic goals for outcome. PMID- 9329293 TI - Sex differences in intelligence. Implications for education. AB - Sex differences in intelligence is among the most politically volatile topics in contemporary psychology. Although no single finding has unanimous support, conclusions from multiple studies suggest that females, on average, score higher on tasks that require rapid access to and use of phonological and semantic information in long-term memory, production and comprehension of complex prose, fine motor skills, and perceptual speed. Males, on average, score higher on tasks that require transformations in visual-spatial working memory, motor skills involved in aiming, spatiotemporal responding, and fluid reasoning, especially in abstract mathematical and scientific domains. Males, however, are also over represented in the low-ability end of several distributions, including mental retardation, attention disorders, dyslexia, stuttering, and delayed speech. A psychobiosocial model that is based on the inextricable links between the biological bases of intelligence and environmental events is proposed as an alternative to nature-nurture dichotomies. Societal implications and applications to teaching and learning are suggested. PMID- 9329294 TI - Intelligence and lifelong learning. What's new and how can we use it? AB - The field of intelligence as applied to lifelong learning has some new ideas about what intellectual abilities are and how they can be measured. The field also has come to a realization of what some of the limitations are both of conventional tests and even of new tests that are starting to emerge. What does the future hold for this field? It is in our hands, as a field, to decide. PMID- 9329295 TI - Improving the health of the world's poor. PMID- 9329296 TI - The trouble with bone allograft. PMID- 9329297 TI - Consumer participation in research and health care. PMID- 9329298 TI - Adverse drug reactions: finding the needle in the haystack. PMID- 9329299 TI - China's smoking epidemic grows. PMID- 9329300 TI - French doctors miss out on postmortem examinations. PMID- 9329301 TI - Colombia struggles with health reform. PMID- 9329302 TI - Medicinal marijuana provided at cost price. PMID- 9329303 TI - Randomised, double blind, placebo controlled clinical trial of efficacy of vitamin A treatment in non-measles childhood pneumonia. AB - OBJECTIVE: To evaluate the impact on clinical recovery and severity of the addition of large doses of vitamin A to the standard treatment for childhood pneumonia. DESIGN: A randomised, double blind, placebo controlled trial. SETTING: Study children were recruited at a public hospital in Recife, north east Brazil, an area of marginal vitamin A deficiency. SUBJECTS: 472 children aged 6 to 59 months with clinical diagnosis of pneumonia. INTERVENTIONS: 200,000 IU (infants) or 400,000 IU (1-4 year olds) of vitamin A in oil or similar capsules of placebo divided into two daily oral doses, in addition to the standard treatment. MAIN OUTCOME MEASURES: Duration of the episode and incidence of adverse outcomes. RESULTS: The groups were similar with respect to overall duration of pneumonia and incidence of adverse outcomes. Children who received vitamin A, however, were less likely to have fever by day 3 (P = 0.008) and were 29% less likely to fail to respond to the first line antibiotic (P = 0.054). CONCLUSION: There was little evidence for an effect of vitamin A treatment on the immediate outcome of the pneumonia episode. PMID- 9329304 TI - Acute upper gastrointestinal haemorrhage in west of Scotland: case ascertainment study. AB - OBJECTIVES: To determine the incidence and case fatality of acute upper gastrointestinal haemorrhage in the west of Scotland and to identify associated factors. DESIGN: Case ascertainment study. SETTING: All hospitals treating adults with acute upper gastrointestinal haemorrhage in the west of Scotland. SUBJECTS: 1882 patients aged 15 years and over treated in hospitals for acute upper gastrointestinal haemorrhage during a six month period. MAIN OUTCOME MEASURES: Incidence of acute upper gastrointestinal haemorrhage per 100,000 population per year, and case fatality. RESULTS: The annual incidence was 172 per 100,000 people aged 15 and over. The annual population mortality was 14.0 per 100,000. Both were higher among elderly people, men, and patients resident in areas of greater social deprivation. Overall case fatality was 8.2%. This was higher among those who bled as inpatients after admission for other reasons (42%) and those admitted as tertiary referrals (16%). Factors associated with increased case fatality were age, uraemia, pre-existing malignancy, hepatic failure, hypotension, cardiac failure, and frank haematemesis or a history of syncope at presentation. Social deprivation, sex, and anaemia were not associated with increased case fatality after adjustment for other factors. CONCLUSIONS: The incidence of acute upper gastrointestinal haemorrhage was 67% greater than the highest previously reported incidence in the United Kingdom, which may be partially attributable to the greater social deprivation in the west of Scotland and may be related to the increased prevalence of Helicobacter pylori. Fatality after acute upper gastrointestinal haemorrhage was associated with age, comorbidity, hypotension, and raised blood urea concentrations on admission. Although deprivation was associated with increased incidence, it was not related to the risk of fatality. PMID- 9329306 TI - Electronic monitoring of vaccine cold chain in a metropolitan area. PMID- 9329305 TI - Water fluoridation, tooth decay in 5 year olds, and social deprivation measured by the Jarman score: analysis of data from British dental surveys. AB - OBJECTIVE: To examine the effect of water fluoridation, both artificial and natural, on dental decay, after socioeconomic deprivation was controlled for. DESIGN: Ecological study based on results from the NHS dental surveys in 5 year olds in 1991-2 and 1993-4 and Jarman underprivileged area scores from the 1991 census. SETTING: Electoral wards in three areas: Hartlepool (naturally fluoridated), Newcastle and North Tyneside (fluoridated), and Salford and Trafford (non-fluoridated). SUBJECTS: 5 year old children (n = 10,004). INTERVENTION: Water fluoridation (artificial and occurring naturally). MAIN OUTCOME MEASURE: Ward tooth decay score (score on the "decayed, missing, and filled tooth index" for each electoral ward). RESULTS: Multiple linear regression showed a significant interaction between Jarman score for ward, mean number of teeth affected by decay, and both types of water fluoridation. This confirms that the more deprived an area, the greater benefit derived from fluoridation, whether natural or artificial (R2 = 0.84, P < 0.001). At a Jarman score of zero (national mean score) there was a predicted 44% reduction in decay in fluoridated areas, increasing to a 54% reduction in wards with a Jarman score of 40 (very deprived). The area with natural fluoridation (at a level of 1.2 parts per million-higher than levels in artificially fluoridated areas) had a 66% reduction in decay, with a 74% reduction in wards with a Jarman score of 40. CONCLUSION: Tooth decay is confirmed as a disease associated with social deprivation, and the more socially deprived areas benefit more from fluoridation. Widespread water fluoridation is urgently needed to reduce the "dental health divide" by improving the dental health of the poorer people in Britain. PMID- 9329307 TI - Reporting of adverse drug reactions by hospital pharmacists: pilot scheme. PMID- 9329308 TI - Prescribing behaviour in clinical practice: patients' expectations and doctors' perceptions of patients' expectations--a questionnaire study. AB - OBJECTIVES: To examine the effect of patients' expectations for medication and doctors' perceptions of patients' expectations on prescribing when patients present with new conditions. DESIGN: Questionnaire study of practitioners and patients. SETTING: General practice in Newcastle, Australia. SUBJECTS: 22 non randomly selected general practitioners and 336 of their patients with a newly diagnosed medical condition. MAIN OUTCOME MEASURES: Prescription of medication and expectation of it. RESULTS: Medication was prescribed for 169 (50%) patients. After controlling for the presenting condition, patients who expected medication were nearly three times more likely to receive medication (odds ratio = 2.9, 95% confidence interval 1.3 to 6.3). When the general practitioner thought the patient expected medication the patient was 10 times more likely to receive it (odds ratio = 10.1, 5.3 to 19.6). A significant association existed between patients' expectation and doctors' perception of patients' expectation (chi 2 = 52.0, df = 4, P = 0.001). For all categories of patient expectation, however, patients were more likely to receive medication when the practitioner judged the patient to want medication than when the practitioner ascribed no expectation to the patient. CONCLUSIONS: Although patients brought expectations to the consultation regarding medication, the doctors' opinions about their expectations were the strongest determinants of prescribing. PMID- 9329310 TI - Fortnighly review. Stress, the brain, and mental illness. PMID- 9329309 TI - The diabetes audit and research in Tayside Scotland (DARTS) study: electronic record linkage to create a diabetes register. DARTS/MEMO Collaboration. AB - OBJECTIVES: To identify all patients with diabetes in a community using electronic record linkage of multiple data sources and to compare this method of case ascertainment with registers of diabetic patients derived from primary care. DESIGN: Electronic capture-recapture linkage of records included data on all patients attending hospital diabetes clinics, all encashed prescriptions for diabetes related drugs and monitoring equipment, all patients discharged from hospital, patients attending a mobile unit for eye screening, and results for glycated haemoglobin and plasma glucose concentrations from the regional biochemistry database. Diabetes registers from primary care were from a random sample of eight Tayside general practices. A detailed manual study of relevant records for the 35,144 patients registered with these eight general practices allowed for validation of the case ascertainment. SETTING: Tayside region of Scotland, population 391,274 on 1 January 1996. MAIN OUTCOME MEASURES: Prevalence of diabetes; population of patients identified by different data sources; sensitivity and positive predictive value of ascertainment methods. RESULTS: Electronic record linkage identified 7596 diabetic patients, giving a prevalence of known diabetes of 1.94% (0.21% insulin dependent diabetes, 1.73% non-insulin dependent): 63% of patients had attended hospital diabetes clinics, 68% had encashed diabetes related prescriptions, 72% had attended the mobile eye screening unit, and 48% had biochemical results diagnostic of diabetes. A further 701 patients had isolated hyperglycaemia (plasma glucose > 11.1 mmol/l) but were not considered diabetic by general practitioners. Validation against the eight general practices (636 diabetic patients) showed electronic linkage to have a sensitivity of 0.96 and a positive predictive value of 0.95 for ascertainment of known diabetes. General practice lists had a sensitivity of 0.91 and a positive predictive value of 0.98. CONCLUSIONS: Electronic record linkage was more sensitive than general practice registers in identifying diabetic subjects and identified an additional 0.18% of the population with a history of hyperglycaemia who might warrant screening for undiagnosed diabetes. PMID- 9329311 TI - ABC of mental health. Mental health on the margins. PMID- 9329312 TI - How to read a paper. Papers that report diagnostic or screening tests. PMID- 9329313 TI - Intensive insulin treatment after acute myocardial infarction in diabetes mellitus. Evidence exists from study of non-insulin dependent diabetes in Japan. PMID- 9329314 TI - Intensive insulin treatment after acute myocardial infarction in diabetes mellitus. Factors other than continued use of subcutaneous insulin may be important. PMID- 9329315 TI - Intensive insulin treatment after acute myocardial infarction in diabetes mellitus. Intensive insulin regimens in primary prevention should be assessed. PMID- 9329316 TI - Breast cancer risk with cyst type in cystic disease of the breast. Consistency of cyst type needs to be known. PMID- 9329317 TI - Breast cancer risk with cyst type in cystic disease of the breast. Larger study found no association between cyst type and breast cancer. PMID- 9329318 TI - General practitioners' workload in primary care led NHS. Workload for chronic disease management has increased substantially. PMID- 9329319 TI - General practitioners' workload in primary care led NHS. Policies of comprehensive anticipatory care require extra doctors and staff. PMID- 9329320 TI - General practitioners' workload in primary care led NHS. Practice's consultation rates have increased by three quarters in past 25 years. PMID- 9329321 TI - Data on health economics of pulmonary rehabilitation programmes are needed. PMID- 9329322 TI - GPs' perceptions of tolerability of selective serotonin reuptake inhibitors and tricyclic antidepressants. Research into long term use is needed. PMID- 9329323 TI - GPs' perceptions of tolerability of selective serotonin reuptake inhibitors and tricyclic antidepressants. Clinical assessments are liable to bias. PMID- 9329324 TI - GPs' perceptions of tolerability of selective serotonin reuptake inhibitors and tricyclic antidepressants. Analysis should discriminate between newer and older tricyclic antidepressants. PMID- 9329325 TI - Anonymity for unrelated bone marrow donors should remain. PMID- 9329326 TI - Diagnosing and managing polymyalgia rheumatica and temporal arteritis. Sensitivity of temporal artery biopsy varies with biopsy length and sectioning strategy. PMID- 9329327 TI - Diagnosing and managing polymyalgia rheumatica and temporal arteritis. Oral prednisolone 40 mg daily is not adequate for temporal arteritis once vision is affected. PMID- 9329328 TI - Diagnosing and managing polymyalgia rheumatica and temporal arteritis. Urgency in giving steroids in giant cell arteritis is still not widely appreciated. PMID- 9329329 TI - Diagnosing and managing polymyalgia rheumatica and temporal arteritis. Repeated measurements of erythrocyte sedimentation rate are not efficient use of time or resources. PMID- 9329330 TI - Diagnosing and managing polymyalgia rheumatica and temporal arteritis. Patients starting steroids should be given advice on risk of osteoporosis. PMID- 9329331 TI - Doctors should be trained in lifting patients. PMID- 9329332 TI - Cognitive dysfunction may complicate assessment of pain in elderly patients. PMID- 9329333 TI - EU directive on bovine spongiform encephalopathy will not affect drugs. PMID- 9329334 TI - Obstructive sleep apnoea. Authors' reply. PMID- 9329335 TI - The role of radiotherapy in acromegaly. PMID- 9329336 TI - Pituitary irradiation is ineffective in normalizing plasma insulin-like growth factor I in patients with acromegaly. AB - Pituitary irradiation suppresses GH hypersecretion in patients with acromegaly. Within 10 yr after radiotherapy, up to 80% of patients achieve plasma GH levels below 5 micrograms/L. Whether this is sufficient to normalize plasma insulin-like growth factor I (IGF-I) levels, is unknown. We examined the effect of radiotherapy on plasma IGF-I concentrations in patients with acromegaly. We reviewed hospital charts of 140 patients with acromegaly seen in our institution between 1975 and 1996. Data on plasma GH and IGF-I were extracted and tabulated longitudinally together with the information about the concomitant medical therapy. We included data from the patients who received radiotherapy as a part of their treatment and whose IGF-I was monitored for more than 1 yr afterward. To avoid the potential bias, the data for patients who were referred to us for medical therapy, having failed radiation elsewhere, were excluded. A total of 38 datasets were submitted for the final analysis. The average follow-up was 6.8 +/- 0.8 yr (range, 1-19). Only 2 patients achieved age- and sex-adjusted normal IGF-I levels while off medical therapy. Noncured patients had a mean plasma GH level of 4.6 +/- 1.1 micrograms/L but still elevated plasma IGF-I levels (219 +/- 26% of the upper normal limit) at the last follow-up visit. A random GH concentration below 1.5 micrograms/L was associated with a pathologically high plasma IGF-I concentration in 43% of instances. Radiotherapy appears to be ineffective in normalizing plasma IGF-I levels in acromegaly. A multicenter study to reevaluate the future use of this modality in patients with acromegaly is warranted. PMID- 9329337 TI - Increase in plasma thyrotropin levels in hypothyroid patients during treatment due to a defect in the commercial preparation. AB - Around mid-1995, the Molecular Endocrinology Laboratory of the Regional Hospital (Malaga, Spain) began detecting an increase in TSH levels in the serum of patients under study to control the treatment of hypothyroidism with levothyroxine. Over a period of 5 months, of a total of 467 hypothyroid patients treated with Levothyroid, 53% had TSH levels higher than 6 microU/mL. The reliability of the biochemical results was verified by duplicating 56 randomly chosen samples from all those with high TSH levels and by an external control performed in four different laboratories. The amount of levothyroxine in the tablets was analyzed by RIA, high performance liquid chromatography, and their iodine contents. The lowest levels of levothyroxine found in the 50 micrograms Levothyroid tablets were those determined by RIA, with a mean value of 32.3 micrograms, resulting in a 35.3% loss of activity. The mean value of levothyroxine found in these same tablets by high performance liquid chromatography was 39.3 micrograms, amounting to a 21.3% loss in activity. The iodine showed no significant loss in these tablets, with a mean experimental value of 48 micrograms. The commercial laboratory withdrew lot J from the market, the one in which these deficiencies were found. PMID- 9329338 TI - Long-term outcome in children and adolescents after transsphenoidal surgery for Cushing's disease. AB - Cushing's disease refers specifically to an ACTH-producing pituitary adenoma that stimulates excess cortisol production. Transsphenoidal surgery is the treatment of choice in children and adolescents, but disparate cure rates have been reported, ranging from 50-98%. The discrepancies in cure rate are due primarily to the technical success of the surgery and the length and method of follow-up. We studied 42 consecutive children and adolescents (age, < or = 18 yr) who underwent transsphenoidal exploration for the primary treatment of Cushing's disease at University of California-San Francisco from 1974-1993. Only 7 patients had persistent disease, defined as evidence of Cushing's disease within 6 months of surgery, yielding an initial remission rate of 83%. We comprehensively evaluated 26 of the 35 patients who experienced an initial remission, including testing of the ACTH-adrenocortical axis. The mean duration of follow-up is 7.2 yr (range, 1.5-13.6 yr). Seven experienced a relapse of Cushing's disease, yielding a net remission rate of 73%. Relapses occurred an average of 4.2 yr postoperatively (range, 0.75-6.2 yr). Five patients experienced relapse within 5 yr of surgery, whereas 2 relapsed more than 5 yr postoperatively. Repeat transsphenoidal surgery was performed in 8 patients with persistent or recurrent disease, and 6 of these remain in remission. Low serum or urinary cortisol measurements within the first post-operative week predicted remission of Cushing's disease, but were not necessarily predictive of long-term cure. Hypercortisolism had significant effects on bone metabolism, as reflected by both diminished bone density in the majority of patients examined and decreased growth rate. Both parameters improved after surgical care, although they did not fully normalize. We conclude that transsphenoidal surgery is a safe and effective treatment for pediatric Cushing's disease, but long-term surveillance is necessary to detect possible recurrences. PMID- 9329339 TI - Effects of leuprolide-induced hypogonadism and testosterone replacement on sleep, melatonin, and prolactin secretion in men. AB - The possible role of gonadal steroids in regulating sleep and circadian rhythms in humans has received relatively little attention despite the importance of the topic to several clinical syndromes. Pharmacologically induced hypogonadism, with and without gonadal steroid replacement, provides an opportunity to examine these questions within a controlled experimental design. We used leuprolide acetate, with and without testosterone replacement, to study the role of testosterone in the regulation of sleep and of melatonin, PRL, and TSH secretion in men. Results from 10 men revealed significant decreases in 24-h PRL levels and in the percentage and time of stage 4 sleep in the hypogonadal state compared with testosterone replacement. There were no differences in melatonin or TSH secretion or in the timing or duration of sleep between the two hormonal conditions. These results indicate that testosterone has relatively specific and discrete effects on sleep and hormonal rhythms in men. PMID- 9329340 TI - Regular exercise and the age-related decline in resting metabolic rate in women. AB - A low resting metabolic rate (RMR) is a risk factor for future weight gain. We tested the hypothesis that the age-related decline in RMR in sedentary women is not observed in women who exercise regularly. Sixty-five healthy, weight-stable women, aged 21-35 or 50-72 yr, were studied: 12 premenopausal and 15 postmenopausal sedentary women, 13 premenopausal and 15 postmenopausal distance runners, and 10 endurance-trained postmenopausal swimmers. RMR was measured by indirect calorimetry (ventilated hood system) after an overnight fast, and values were adjusted for fat mass and fat-free mass (RMRadj). The RMRadj was approximately 10% lower in the postmenopausal vs. premenopausal sedentary women (52 +/- 2 vs. 57 +/- 2 Cal/h; P < 0.002). In contrast, RMRadj was not significantly different in the premenopausal (59 +/- 2 Cal/h) and postmenopausal (57 +/- 1 Cal/h) distance runners. The postmenopausal swimmers had a RMRadj (57 +/- 2 Cal/h) identical to that of the postmenopausal runners, suggesting a generalized influence of the endurance exercise-trained state in postmenopausal women. Group differences in RMRadj were not associated with differences in total energy intake or composition or with plasma concentrations of norepinephrine, T3, or T4. However, maximal oxygen consumption (aerobic fitness) accounted for 35% of the individual variance in RMRadj in the overall population (r = 0.59; P < 0.001). Our results are consistent with the concept that the age-related decline in RMR in sedentary women is not observed in women who regularly perform endurance exercise. The elevated level of RMR observed in middle-aged and older exercising women may play a role in their lower levels of body weight and fatness compared to those in sedentary women. PMID- 9329341 TI - Testosterone treatment in adolescents with delayed puberty: changes in body composition, protein, fat, and glucose metabolism. AB - Previously, we demonstrated decreased protein breakdown and insulin resistance in pubertal adolescents compared with prepubertal children. Puberty-related increases in sex steroids and/or GH could be potentially responsible. In the present study, the effects of 4 months of testosterone enanthate (50 mg in every 2 weeks) on body composition, protein, fat, and glucose metabolism and insulin sensitivity were evaluated in adolescents with delayed puberty. Body composition was assessed by H218O-dilution principle. Protein breakdown, oxidation, and synthesis were measured during primed constant infusion of [1-13C]leucine. Whole body lipolysis was measured during primed constant infusion of [2H5]glycerol. Insulin action in suppressing proteolysis and lipolysis and stimulating glucose disposal was assessed during a stepwise hyperinsulinemic (10 and 40 mU-m2.min) euglycemic clamp. Fat and glucose oxidation rates were calculated from indirect calorimetry measurements. After 4 months of testosterone treatment, height, weight, and fat free mass (FFM) increased and fat mass, percent body fat, plasma cholesterol, high- and low-density lipoproteins, and leptin levels decreased significantly. Whole-body proteolysis and protein oxidation were lower after testosterone treatment (proteolysis, 0.49 +/- 0.03 vs 0.54 +/- 0.04 g.h.kg FFM, P = 0.032; oxidation, 0.05 +/- 0.01 vs. 0.09 +/- 0.01 g.h.kg FFM, P = 0.015). Protein synthesis was not different, and resting energy expenditure was not different. Total body lipolysis was not affected by testosterone treatment, however, fat oxidation was higher after testosterone (pre-: 2.4 +/- 0.7 vs. post : 3.5 +/- 0.7 mumol.kg.min, P = 0.031). During the 40 mU.m2.min hyperinsulinemia, insulin sensitivity of glucose metabolism was not affected with testosterone therapy (59.1 +/- 8.8 vs. 57.1 +/- 8.2 mumol.kg.min per muU/mL). However, metabolic clearance rate of insulin was higher posttestosterone (13.6 +/- 1.1 vs. 16.7 +/- 0.8 mL.kg.min, P = 0.004). In conclusion, after 4 months of low-dose testosterone treatment in adolescents with delayed puberty 1) FFM increases and fat mass and leptin levels decrease; 2) postabsorptive proteolysis and protein oxidation decrease; 3) fat oxidation increases; and 4) insulin sensitivity in glucose metabolism does not change, whereas insulin clearance increases. These longitudinal observations are in agreement with our previous cross-sectional studies of puberty and demonstrate sparing of protein breakdown of approximately 1.2 g.kg.day FFM, wasting of fat mass, but no change in insulin sensitivity after short periods of low-dose testosterone supplementation. PMID- 9329342 TI - Insulin resistance does not change the ratio of proinsulin to insulin in normal volunteers. AB - Plasma glucose, insulin, and proinsulin concentrations were measured before and after an oral glucose challenge in 57 nondiabetic individuals. In addition, insulin-mediated glucose disposal was estimated by determining the steady state plasma glucose (SSPG) concentration after a 180-min iv infusion of somatostatin, insulin, and glucose. The plasma glucose concentration after oral glucose administration was used to divide the population into those with normal (n = 36) or impaired glucose tolerance (IGT; n = 21), and the 36 normal glucose-tolerant individuals were further subdivided into an insulin-sensitive (SSPG, < 9.0 mmol/L; n = 15) and an insulin-resistant (SSPG, > 10 mmol/L; n = 21) group. Fasting and postglucose load insulin concentrations were similar in the normal glucose-tolerant insulin-resistant and IGT groups, but were significantly higher (P < 0.02- < 0.001) than those in normal glucose-tolerant insulin-sensitive individuals. Fasting proinsulin concentrations were also higher (P < 0.002) in the normal glucose-tolerant insulin-resistant (15.1 +/- 1.5 pmol/L) and IGT (15.8 +/- 1.8 pmol/L) groups compared to those in normal glucose-tolerant insulin sensitive volunteers (9.3 +/- 1.2 pmol/ L). However, the ratio of fasting proinsulin to insulin was identical in all three groups (0.12). When the three groups were combined, significant relationships (P < 0.001) existed between SSPG (degree of insulin resistance) and both fasting proinsulin (r = 0.59) and insulin (r = 0.66) concentrations, but not with the ratio of proinsulin to insulin (r = 0.03). These results demonstrate that fasting proinsulin and insulin concentrations are increased in insulin-resistant, nondiabetic subjects, and the more insulin resistant, the greater the increase. In contrast, the ratio of proinsulin to insulin did not vary as a function of insulin resistance. Thus, neither insulin resistance nor the need to secrete more insulin to maintain glucose tolerance necessarily leads to abnormal insulin processing by the beta cell. PMID- 9329343 TI - Spontaneous nocturnal growth hormone secretion in anorexia nervosa. AB - In anorexia nervosa, serum GH levels are increased under basal conditions and respond abnormally to provocative stimuli. We report here, for the first time, an analysis of pulsatile GH secretion in these patients performed by Cluster algorithm. Seven anorectic and six normal weight, healthy women underwent serial blood sampling at 20-min intervals form 2030-0830 h for GH estimation. The total area under the curve (AUC; micrograms per L/min) was elevated 4-fold in anorectic patients compared to controls (4743.0 +/- 1520.09 vs. 1148.6 +/- 519.27; P < 0.01), largely due to an increase in the non-pulsatile fraction (3212.5 +/- 990.45 vs. 378.7 +/- 123.27; P < 0.01). Accordingly, the valley mean value was higher in anorectic than in control subjects (5.9 +/- 2.25 vs. 1.0 +/- 1.30 micrograms/L; P < 0.01). Furthermore, pulsatile AUC was also greater in anorectic patients (1530.4 +/- 654.72 vs. 769.8 +/- 404.02; P < 0.01) due to a significant increase in GH peak frequency (5.0 +/- 0.81 vs. 3.0 +/- 0.89; P < 0.01). No correlations were observed in these patients between body mass index and any of the parameters of spontaneous GH release, whereas a positive correlation was found between insulin-like growth factor I levels and pulsatile AUC (r2 = 0.583; P < 0.05), peak height (r2 = 0.743; P = 0.01), peak increment (r2 = 0.801; P < 0.01), and GH valley mean (r2 = 0.576; P < 0.05). In conclusion, it appears that the enhanced GH secretion in anorexia nervosa is the result of an increased frequency of secretory pulses superimposed on enhanced tonic GH secretion. Although this latter is consistent with a reduction of hypothalamic SRIH tone, the former may be accounted for by an increased number of GHRH discharges. Considering that in normal weight and obese subjects parameters of GH release are negatively correlated with adiposity indexes, the lack of such a negative correlation in our patients suggests that the enhancement of spontaneous GH release in anorectic patients is not merely the consequence of malnutrition dependent impairment of insulin-like growth factor I production, but reflects a more complex hypothalamic dysregulation of GH release. PMID- 9329344 TI - Robust leptin secretory responses to dexamethasone in obese subjects. AB - Although leptin reverses obesity in rodents, its function and regulation in humans are unknown. Glucocorticoids have been reported to stimulate leptin production in both rodents and humans, but data assessing the effect of obesity on dynamic leptin secretory responses are unavailable. We, therefore, studied 52 lean and obese subjects [20 men and 32 women; aged 19-84 yr; body mass index (BMI) range, 16-47 kg/m2] randomized to treatment with dexamethasone (total dose, 10 mg/4 days) or placebo. Compared with placebo, dexamethasone increased (P = 0.0001) plasma leptin levels by 64-111% above baseline values within 2-4 days. The increases occurred in all ages, showed no sexual dimorphism, and were particularly robust in obese subjects. After dexamethasone treatment, significant interactions were observed between the change in plasma leptin and BMI (P = 0.0001), baseline plasma leptin (P = 0.0006) and plasma dexamethasone levels (P = 0.04), but not age (P = 0.28); an apparent interaction with plasma insulin no longer was significant after controlling for BMI. These results confirm dexamethasone-induced hyperleptinemia in humans and further demonstrate that the response is not defective in obesity. PMID- 9329346 TI - Leptin is inversely related to age at menarche in human females. AB - Over the last century there has been a trend toward an earlier onset of menarche attributed to better nutrition and body fatness. With the discovery of the obesity gene and its product, leptin, we reexamined this hypothesis from a new perspective. As delayed menarche and leanness are considered risk factors for osteoporosis, we also evaluated the relation between leptin and bone mass. Body composition and serum leptin levels were measured, and the timing of menarche was recorded in 343 pubertal females over 4 yr. Body composition was measured by dual x-ray absorptiometry, and leptin by a new RIA. All participants were premenarcheal at baseline (aged 8.3-13.1 yr). Leptin was strongly associated with body fat (r = 0.81; P < 0.0001) and change in body fat (r = 0.58; P < 0.0001). The rise in serum leptin concentration up to the level of 12.2 ng/mL (95% confidence interval, 7.2-16.7) was associated with the decline in age at menarche. An increase of 1 ng/mL in serum leptin lowered the age at menarche by 1 month. A serum leptin level of 12.2 ng/mL corresponded to a relative percent body fat of 29.7%, a body mass index of 22.3, and-body fat of 16.0 kg. A gain in body fat of 1 kg lowered the timing of menarche by 13 days. Leptin was positively related to bone area (r = 0.307; P < 0.0001) and change in bone area (r = 0.274; P < 0.0001). A critical blood leptin level is necessary to trigger reproductive ability in women, suggesting a threshold effect. Leptin is a mediator between adipose tissue and the gonads. Leptin may also mediate the effect of obesity on bone mass by influencing the periosteal envelope. This may have implications for the development of osteoporosis and osteoarthritis. PMID- 9329345 TI - Decreased insulin sensitivity and compensatory hyperinsulinemia after hormone treatment in children with short stature. AB - To assess the effects of GH treatment on carbohydrate and protein metabolism, we studied eight patients with short stature before and after the commencement of GH treatment. The hyperglycemic clamp procedure was employed to produce a hyperglycemic stimulus of 50 mg/dL above fasting levels for 120 min. These patients were then treated with 0.3 mg/kg. week GH for 6 months, after which they were restudied. Patients were compared to eight healthy control children matched for age, sex, and Tanner stage. Fasting plasma glucose did not change significantly, but fasting plasma insulin levels were higher after GH therapy (P < 0.005). Despite identical glucose increments during the glucose clamp procedure, both first, and second phase insulin responses were markedly greater after instituting GH treatment. Even in the face of higher mean plasma insulin levels after GH treatment, the rate of insulin-stimulated glucose metabolism did not differ during the last 60 min of both studies. Hence, the rate of insulin stimulated glucose metabolism/mean plasma insulin ratio (an index of insulin sensitivity) was sharply reduced after GH treatment (P < 0.01). During the clamp, the fall in circulating branched chain amino acid levels was significantly greater after GH therapy (P < 0.02). We conclude that glucose-stimulated insulin responses are increased in short children treated with GH and that such hyperinsulinemic responses compensate for reductions in insulin sensitivity. The compensatory hyperinsulinemic responses induced by GH therapy may serve a beneficial role by augmenting insulin's anabolic effects on protein metabolism. PMID- 9329347 TI - A novel missense mutation in codon 218 of the albumin gene in a distinct phenotype of familial dysalbuminemic hyperthyroxinemia in a Japanese kindred. AB - Familial dysalbuminemic hyperthyroxinemia (FDH) is the most common cause of inherited euthyroid hyperthyroxinemia in Caucasians. To our knowledge, no such documentation on Asians exists. Six of 8 members of a 3-generation Japanese family were found by us to carry the FDH phenotype. Serum total T4 levels ranged from 1763.2-2741.3 nmol/L (normal range, 65.6-164.7), serum total T3 levels ranged from 2.73-5.62 nmol/L (normal range, 1.47-2.95), and rT3 levels ranged from 1.08-2.52 nmol/L (normal range, 0.22-0.60). In the proband, the majority of [125I]T4 in serum T4-binding proteins was distributed in albumin fractions, and the isolated albumin had an increased affinity for T4. A guanine to cytosine transition in the second nucleotide of codon 218, resulting in replacement of normal arginine with proline, was detected in 1 of 2 alleles in all 5 subjects of the family with FDH. In all FDH-affected Caucasian subjects from 10 unrelated families with a moderate increase in serum T4, the guanine to adenine transition was demonstrated at the same position of the albumin gene as noted in our patients, but histidine, the replacement amino acid, differed from proline noted in our FDH Japanese subjects. It would thus appear that FDH has ethnic variations. PMID- 9329348 TI - Cortisol reduces hippocampal glucose metabolism in normal elderly, but not in Alzheimer's disease. AB - Glucocorticoids are known to play a role in the regulation of peripheral glucose mobilization and metabolism. Although several animal studies have shown that hippocampal glucose metabolism is reduced acutely and chronically by the action of corticosterone and that excess glucocorticoids are harmful to hippocampal neurons, little is known about the central effects of glucocorticoids in the human. In this study we examined the brain glucose utilization (CMRglu) response to hydrocortisone (cortisol) in seven normal elderly and eight Alzheimer's disease (AD) patients. On 2 separate days, immediately after the administration of a bolus of either 35 mg hydrocortisone or placebo, we administered 2-deoxy-2 [18F]fluoro-D-glucose. After a 35-min radiotracer uptake period, positron emission tomography (PET) images were collected. PET CMRglu images were analyzed using two methods: an image transformation that allowed analyses across cases on a voxel by voxel basis, and an anatomically based region of interest method that used coregistered magnetic resonance imaging scans. Both image analysis methods yielded similar results, identifying relative to placebo, a specific hippocampal CMRglu reduction in response to the hydrocortisone challenge that was restricted to the normal group. The region of interest technique showed CMRglu reductions of 16% and 12% in the right and left hippocampi, respectively. Blood collected during the PET scans showed, for the normal group, a rise in plasma glucose levels, starting approximately 25 min after hydrocortisone administration. The AD group did not show this effect. Baseline cortisol was elevated in the AD group, but the clearance of hydrocortisone was not different between the groups. In conclusion, these data show that among normal individuals in the presence of a pharmacological dose of cortisol, the glucose utilization of the hippocampus is specifically reduced, and serum glucose levels increase. Based in part on other studies, we offer the interpretation that glucocorticoid-mediated regulation of glucose transport is altered in AD, and this may underlie both the hippocampal insensitivity to cortisol and the failure in these patients to mount a peripheral glucose response. As our findings could reflect an altered state of the AD patients, we interpret our results as preliminary with respect to evidence for metabolic abnormalities in AD. The results suggest the continued study of the hydrocortisone challenge as a test of hippocampal responsivity. PMID- 9329349 TI - Glucocorticoids and the immune function in the human immunodeficiency virus infection: a study in hypercortisolemic and cortisol-resistant patients. AB - Immunological studies in human immunodeficiency virus (HIV)-positive patients suggest that the disease progression is accompanied by a defective production of type 1 cytokines (interleukin-2 (IL-2) and IL-12], an increased production of type 2 cytokines (IL-4, IL-6, and IL-10), and an increased production of IgE. HIV infection is also associated with activation of the hypothalamo-pituitary-adrenal axis function and increased plasma and urinary cortisol concentrations. As cortisol is involved in the physiological regulation of cytokines, a study was conducted to examine cytokine patterns in two groups of hypercortisolemic patients, one with normal sensitivity to glucocorticoids and the other with glucocorticoid resistance. Ten HIV-infected patients with normal receptor affinity to glucocorticoids (AIDS-C), 10 HIV-infected patients with low receptor affinity to glucocorticoids (AIDS-GR), and 20 healthy subjects were studied. Receptor characteristics of peripheral blood mononuclear cells were evaluated by [3H]dexamethasone binding. Serum cortisol and urinary free cortisol were measured by RIA. Serum ACTH and IgE were measured by immunoradiometric assay, and IL-2, IL 4, and IL-10 cytokines and interferon-gamma were measured by enzyme-linked immunosorbent assay. AIDS-C patients showed low IL-2 and high IL-4, IL-10, and IgE concentratios; conversely, AIDS-GR patients showed high IL-2 and low IL-4 and IgE concentrations. Thus, in HIV infection, elevated cortisol levels suppress cell-mediated immunity and stimulate humoral immunity, whereas this response is not detected in cortisol-resistant patients. These findings indicate that cortisol and its receptors are critically involved in the regulation of immune function in HIV infection. PMID- 9329350 TI - Thyroid function in Rubinstein-Taybi syndrome. AB - Rubinstein-Taybi syndrome (RTS) is a genetic syndrome characterized by broad thumbs and halluces, growth retardation, mental retardation, and craniofacial abnormalities. This condition recently was found to be caused by mutations in the gene encoding cAMP response element-binding protein (CREB)-binding protein. As CREB-binding protein has been shown to be a critical coactivator for thyroid hormone receptors, it is plausible that RTS would be characterized by thyroid hormone resistance. In fact, features of RTS, such as mental retardation and short stature, are consistent with thyroid hormone deficiency or resistance. To assess the function of the thyroid axis in RTS, free T4 and TSH were measured in 12 subjects with this syndrome. The free T4 level was normal in all 12 (mean +/- SD, 0.97 +/- 0.20 ng/dL; normal range, 0.73-1.79), as was the TSH level (2.24 +/- 0.87 microU/mL; normal range, 0.3-6.5). Thus, overt thyroid hormone resistance does not appear to be a typical feature of RTS. PMID- 9329351 TI - Reversal of the sex difference in serum leptin levels upon cross-sex hormone administration in transsexuals. AB - Women have higher circulating leptin levels than men. This sex difference is not simply explained by differences in the amount of body fat and is possibly influenced by their different sex steroid milieus. This prompted us to study prospectively the effects of cross-sex steroid hormones on serum leptin levels in 17 male to female transsexuals and 15 female to male transsexuals. Male to female transsexuals were treated with 100 micrograms ethinyl estradiol and 100 mg cyproterone acetate (antiandrogen) daily, and female to male transsexuals received testosterone esters (250 mg/2 weeks, im). Before and after 4 and 12 months of cross-sex hormone treatment, serum leptin levels and measures of body fatness were assessed. Before treatment, female subjects had higher serum leptin levels than male subjects independently of the amount of body fat (P < 0.01). Cross-sex hormone administration induced a reversal of the sex difference in serum leptin levels. Estrogen treatment in combination with antiandrogens in male subjects increased median serum leptin levels from 1.9 ng/mL before treatment to 4.8 ng/mL after 4 months and 5.5 ng/mL after 12 months of treatment (P < 0.0001). Testosterone administration in female subjects decreased median serum leptin levels from 5.6 to 2.6 ng/mL after 4 months and to 2.5 ng/mL after 12 months (P < 0.0001). Analysis of covariance revealed that the changes in serum leptin levels were independent of changes in body fatness in both groups (P < 0.01). In conclusion, these results indicate that sex steroid hormones, in particular testosterone, play an important role in the regulation of serum leptin levels. The prevailing sex steroid milieu, not genetic sex, is a significant determinant of the sex difference in serum leptin levels. PMID- 9329352 TI - Liposomal thyroxine: a noninvasive model for transplacental fetal therapy. AB - Drugs that cross the placenta sparingly are currently given directly to the fetus by invasive procedures. We investigated whether anionic small unilamellar (SUV) liposomes of different lipid compositions enhanced the transfer and uptake of T4 in an in vitro model of perfused human term placenta. T4-encapsulated anionic liposomes were prepared using lecithin (F-SUV) or distearoyl phosphatidylcholine (S-SUV) with cholesterol and dicetylcholine. The size distribution, encapsulation efficiency, and stability were determined in blood-based media. The transfer kinetics of free and liposomally encapsulated T4 were studied in a dually perfused isolated lobule of human term placenta, with creatinine and liposomal carboxyfluorescein as marker substances. Concentrations of T4 and rT3 were measured by RIA. T4 crossed the placenta sparingly (1.9 +/- 0.5%) because it was metabolized to rT3 (9.2 +/- 1.3%). Transplacental transfer of T4 was significantly increased by F-SUV (15.8 +/- 2.1%; P < 0.001) and S-SUV liposomes (7.1 +/- 1.2%; P < 0.001), with a concomitant decrease in fetal rT3 levels (P < 0.001). Placental uptake of F-SUV (13.5 +/- 2.0%; P < 0.001) was greater than that of S-SUV liposomes (6.7 +/- 0.8%; P < 0.001). Our data suggest that anionic liposomes increase transplacental transfer of T4. If confirmed in vivo, liposomes may provide an alternative noninvasive method of drug delivery to the fetus. PMID- 9329353 TI - Rapid eye movement sleep correlates with the overall activities of the hypothalamic-pituitary-adrenal axis and sympathetic system in healthy humans. AB - To assess the association of the overall amount of rapid eye movement (REM) sleep and the activities of the hypothalamic-pituitary-adrenal axis and sympathetic system, we performed polysomnography and measured 24-h urinary free cortisol and catecholamine excretion in 21 healthy adults. After an adaptation night, each subject was recorded in the sleep laboratory for 3 consecutive nights while 24-h urine specimens were collected. Urinary free cortisol, epinephrine, dihydroxyphenylglycol, and dihydroxyphenylacetic acid levels were significantly and positively correlated with the average values of percent REM sleep (P < 0.05). There were no correlations between hormone values and REM latency, other variables of REM distribution, or REM density, an index of phasic activity during REM sleep. The positive correlations between stress system activity and REM sleep are consistent with hormonal and sleep alterations in melancholic depression, a state characterized by increased cortisol and catecholamine secretion. PMID- 9329354 TI - Relationship between concentration of serum leptin and fetal growth. AB - The serum leptin concentration reflects the amount of adipose tissue in the body. Although fat deposition in the fetus in the third trimester markedly increases, the role of leptin during pregnancy has not been clarified. In the present study, whether or not the serum leptin concentration correlates with growth in utero was investigated, in addition to how leptin levels change in the first few days after birth. One hundred sixteen Japanese infants were divided into term (n = 91) and preterm groups (n = 25). Term infants were divided into 3 subgroups: birth weight appropriate for gestational age (AGA) (n = 44), birth weight large for gestational age (LGA) (n = 28), and birth weight small for gestational age (SGA) (n = 19). Longitudinal changes in the concentration of serum leptin after birth were examined in 48 infants. The serum leptin concentration was determined by RIA. No significant difference in leptin levels between cord sera and infants' sera obtained within the first 6 h of life (n = 28) was observed. Within the first 6 h of life, the concentration of serum leptin in LGA infants (12.8 +/- 10.2 ng/mL) and SGA infants (1.6 +/- 1.1 ng/mL) was significantly higher and lower, respectively, than that in the AGA infants (4.4 +/- 3.0 ng/mL) (P < 0.01). A significant positive correlation was found between the leptin concentration within 6 h of life and birth body weight (r = 0.59, P < 0.01). After birth, the concentration of leptin in LGA and AGA infants significantly decreased to the level in SGA infants within 72 h [corrected] of delivery (P < 0.05). After 72 h [corrected] of life, no significant differences in the concentration of leptin were observed among the three groups, and low levels continued to 7 days of age. These findings indicate that serum level of leptin correlates with fetal body weight gain. PMID- 9329356 TI - Population-wide evaluation of disease manifestation in relation to molecular genotype in steroid 21-hydroxylase (CYP21) deficiency: good correlation in a well defined population. AB - We report a population-wide analysis of all patients with 21-hydroxylase deficiency (21-OHD) found in Finland, a country with a genetically well defined population, in which the effects of other genetic and environmental factors on the phenotype can be expected to be low. In total, 120 patients were identified, and their clinical status was evaluated. Blood samples for CYP21 genotype determination could be obtained from 78 (65%) patients, and their phenotypes were compared with their genotypes. In general, the severity of gene defects correlated well with clinical expression. All patients carrying mutations with the most drastic effects on enzymatic activity had the salt-wasting form of 21 OHD. The I2 splice mutation, which in some reports has been connected with clinical variation, was constantly associated with severe mineralocorticoid deficiency. However, patients with I172N as the determining mutation expressed a wide spectrum of phenotypes; the variation could not be attributed to additional mutations. Although genetically affected males with the nonclassical form had not been clinically diagnosed, our study suggests that nonclassical 21-OHD is substantially more rare in Finland than elsewhere, as indicated by both clinical evaluation and mutational screening. PMID- 9329355 TI - Growth hormone (GH) replacement reduces total body fat and normalizes insulin sensitivity in GH-deficient adults: a report of one-year clinical experience. AB - The effects of GH replacement on body fat composition and insulin sensitivity were assessed in GH-deficient adults. The patients were randomized into a double blind, placebo-controlled study of human recombinant GH replacement therapy for 6 months (period 1), followed by an open phase of GH for another 6 months (period 2). Anthropometric variables, body fat composition (fat %), and biochemical parameters were measured during the trial. Measurements of in vivo insulin sensitivity were carried out at the commencement of the study and on completion of the trial by modified insulin suppression test. The modified insulin suppression test was performed both in the morning (AM) and in the afternoon (PM) to further evaluate the PM-AM steady-state plasma glucose (SSPG) pattern. We found that the GH-deficient adults had more body fat and were insulin resistant. Significant reduction in fat % and total body fat mass was found in the active arm of period 1 without alteration of body weight. Besides, we demonstrated, for the first time, the GH replacement for 6 months did not alter the insulin sensitivity, but replacement for a longer period (12 months) normalized not only the AM SSPG level but also the PM-AM SSPG pattern. We also found a positive correlation between SSPG (regardless of AM vs. PM) and fat % and total body fat mass. In conclusion, normalization of insulin sensitivity in GH-deficient adults after replacement of GH may be related to the reduction of total body fat. PMID- 9329357 TI - Menstrual bleeding in a female infant with congenital adrenal hyperplasia: altered maturation of the hypothalamic-pituitary-ovarian axis. AB - Vaginal bleeding during the neonatal period is commonly related to the withdrawal of maternal estrogens. Vaginal bleeding has also been reported in female infants with congenital adrenal hyperplasia and has been proposed to be due to a treatment-induced activation of the hypothalamic-pituitary-ovarian axis. We report a female infant with the salt-losing form of congenital adrenal hyperplasia due to 21-hydroxylase deficiency, who had the onset of vaginal bleeding at 3 months of life. Adrenal steroid suppression had been achieved by 2.5 weeks of age. At the time of bleeding, imaging studies revealed an enlarged right ovary with a dominant 3-cm cyst and additional small cysts that had not been seen on the newborn sonogram. The uterus was enlarged and stimulated. Three weeks later (1 week after the cessation of bleeding), repeat ultrasound demonstrated a marked decrease in the size of the right ovary, and the dominant cyst was no longer seen. The patient had a heightened FSH response to GnRH and elevated levels of estradiol for age. At 5 months of age, no further episodes of sustained vaginal bleeding were observed. Repeat hormonal levels were prepubertal, and pelvic sonogram demonstrated no evidence of stimulation. The findings in our patient suggest that a decline in adrenal androgens after glucocorticoid treatment resulted in an increase in gonadotropin levels, which then triggered a transient and augmented end-organ response (menses). Further, we suggest that our infant's hormonal findings may reflect a delay in the timely development of the negative restraint by sex steroids on gonadotropins that is normally observed in infancy. PMID- 9329358 TI - Serum markers of bone and collagen turnover in patients with Cushing's syndrome and in subjects with adrenal incidentalomas. AB - The aim of this study was to assess serum levels of some markers of bone turnover and collagen synthesis in 22 patients with adrenal incidentalomas (AI), a model of silent glucocorticoid excess, and to compare the results with those obtained in 18 patients with Cushing's syndrome (CS). Osteocalcin (BGP), bone isoenzyme of alkaline phsophatase, carboxy-terminal propeptide of type I procollagen, and carboxy-terminal cross-linked telopeptide of type I collagen were measured as biochemical indexes of bone turnover, and amino-terminal propeptide of type III procollagen was determined as an index of collagen synthesis. Two groups of healthy volunteers evenly matched for sex, age, and menstrual status were used for a case-control analysis of AI and CS groups, respectively. Patients with AI showed a slight, albeit significant, reduction in serum BGP and a mild increase in carboxy-terminal cross-linked telopeptide of type I collagen levels compared with controls [median, 6.6 vs. 7.8 ng/mL (P < 0.05) and 4.2 vs. 3.1 micrograms/L (P < 0.01), respectively]. No significant differences were found when comparing the other markers. Patients with CS had BGP, bone isoenzyme of alkaline phosphatase, and amino-terminal propeptide of type III procollagen levels significantly lower than control values [median, 3.0 vs. 7.3 ng/mL (P < 0.0001); 4.4 vs. 11.5 micrograms/L (P < 0.01); 2.2 vs. 4.3 micrograms/L (P < 0.0001), respectively], but no significant difference in the other markers. These results confirm a clear inhibition of osteoblastic activity in CS and could suggest an enhanced bone metabolism in patients with AI. The degree of impairment of bone turnover in patients with AI does not seem enough to recommend surgery (removal of the adrenal adenoma) in the absence of other indications. PMID- 9329359 TI - Effect of octreotide pretreatment on surgical outcome in acromegaly. AB - Pretreatment with octreotide (OCT) in acromegaly has been reported to improve surgical outcome. The objective of this study was to analyze retrospectively the effects of a 3- to 6-month presurgical treatment with OCT in acromegalics focusing on electrocardiographic (ECG) records, blood pressure levels, glucose and lipid profile, tumor size and consistency, easy tumor removal at surgery, and morphological findings at pathology. Fifty-nine patients with acromegaly who were undergoing surgical treatment were studied randomly before surgery; 37 patients were untreated, and 22 were treated with OCT at doses ranging 150-600 micrograms/day for 3-6 months. At study entry, untreated and OCT-treated patients had similar circulating GH and insulin-like growth factor I (IGF-I), glucose, and cholesterol levels as well as prevalence of overt diabetes mellitus, hypertension, and ECG abnormalities. In untreated and OCT-treated patients, respectively, radiological imaging documented microadenoma in 0 and 1, intrasellar macroadenoma in 10 and 6, intra- and suprasellar macroadenoma in 18 and 11, invasive macroadenoma in 9 and 4 patients. Before surgery, serum GH and IGF-I levels significantly decreased in the 22 OCT-treated acromegalics, and in 5 of them, a significant shrinkage was documented. ECG abnormalities disappeared in 7 of 11 (63.6%) OCT-treated patients. In 3 of the 7 patients with diabetes mellitus, treatment with OCT together with low carbohydrate intake normalized blood glucose levels, whereas in 2 patients, insulin could be replaced by oral antidiabetics, and in 2 patients, the insulin dose was reduced. Presurgical blood glucose, total cholesterol and triglyceride levels, as well as systolic (145.2 +/ 3.4 vs. 132.9 +/- 2.5 mm Hg; P < 0.01) and diastolic (94.3 +/- 1.7 vs. 84.3 +/- 1.6 mm Hg; P < 0.001) blood pressure levels were significantly higher in untreated than in OCT-treated patients. Two weeks after surgery, circulating GH and IGF-I levels were normalized in 11 untreated (29.7%) and 12 OCT-treated (54.5%) patients (P < 0.005, by chi 2 test). Macroscopically, no difference was found between untreated and OCT-treated adenomas, whereas at pathology, a significant increases in cellular atypia (31.6% vs. 19.2%; P < 0.05) was found in OCT-treated adenomas. One patients in the untreated group died from cardiorespiratory arrest during the early postoperative period. Finally, the average duration of hospitalization after operation was longer in untreated than in OCT-treated patients (8.6 +/- 0.7 vs. 5.6 +/- 0.5 days). We conclude that a 3- to 6-month treatment with OCT before surgery for GH-secreting adenoma improved clinical conditions and surgical outcome and reduced the duration of hospitalization after operation. PMID- 9329360 TI - Clinical and biochemical features of muscle dysfunction in subclinical hypothyroidism. AB - Alterations in muscle structure and function have been reported in overt hypothyroidism, with decreased activity of enzymes involved in anaerobic and oxidative glucose metabolism. To test whether similar changes in muscle energy metabolism are present in subclinical hypothyroidism (sHT), we studied 12 patients with sHT who complained of mild neuromuscular symptoms. The control group included 10 sex- and age-matched healthy volunteers. Skeletal muscle lactate and pyruvate production were determined in the resting state and during dynamic arm exercise. During exercise, blood lactate was significantly higher in sHT patients than in controls from the third exercise step onward (P = 0.02 at 30%, p = 0.008 at 40%, and P = 0.002 at 50% of maximal voluntary contraction). Moreover, the mean increment in blood lactate during exercise was positively related (r2 = 0.44; P = 0.02) to the duration of sHT, but not to serum levels of TSH, free T3, or free T4. No significant difference was found in blood pyruvate concentrations between the two groups at baseline or during exercise. Thus, the lactate/pyruvate ratio curve paralleled the lactate curve in patients as well as controls. We conclude that muscle energy metabolism is impaired in sHT in rough proportion to the known duration of the disease. Early L-T4 therapy may be useful not only to provide specific treatment for such metabolic changes, but also to avoid progression to frank hypothyroidism. PMID- 9329361 TI - Keratinocyte growth factor expression in the mesenchymal cells of human amnion. AB - Amnion epithelial and mesenchymal cells were separated by differential protease treatment, and the separated cells were maintained in monolayer culture. Keratinocyte growth factor (KGF) messenger RNA (mRNA) was readily detected by Northern analysis of amnion mesenchymal cell total RNA (10 micrograms) but not in amnion epithelial cells. Treatment of the amnion mesenchymal cells in serum-free medium with tetradecanoyl phorbol acetate (1 nM) caused an increase in the level of KGF mRNA. Forskolin treatment also caused an increase in KGF mRNA but not to the levels attained with tetradecanoyl phorbol acetate treatment. Dexamethasone (1 nM) treatment of these cells effected a reduction in the level of KGF mRNA. Prolonged maintenance of mesenchymal cells in serum-free medium also was associated with an increase in the level of KGF mRNA. Treatment with a variety of other agents, viz., interleukin (IL)-1, IL-6 plus or minus IL-6 soluble receptor, IL-11, oncostatin M, epidermal growth factor (EGF), and transforming growth factor-beta and not modify the level of KGF mRNA. Treatment of amnion epithelial cells with KGF caused an increase in the rate of [3H]thymidine incorporation, but the rate of cell replication induced by KGF was less than that induced by treatment with EGF. Transforming growth factor-beta treatment inhibited basal and EGF- and KGF-stimulated amnion epithelial cell replication. The findings of this study are indicative the KGF is expressed in human amnion mesenchymal cells, and that KGF may act on the epithelial cells of this tissue. PMID- 9329363 TI - Leptin concentration in cord blood correlates with intrauterine growth. AB - Leptin is an adipocyte-derived peptide hormone regulating energy balance in experimental animals. Although the physiological function of leptin in humans is still unclear, its secretion is closely related to fat mass in adult humans. To examine how fetal growth correlates with leptin levels at birth, an umbilical cord venous blood sample was obtained at the delivery from 50 term newborn infants. Twenty-eight of the newborn infants had birth weights appropriate for gestational age (AGA; mean +/- SEM, 3362 +/- 90 g; relative birth weight, -0.08 +/- 0.2 SD), 9 were large for gestational age (birth weight, 4655 +/- 165 g; relative birth weight, 3.2 +/- 0.3 SD; P < 0.001 vs. AGA newborn infants), and 13 were small for gestational age (SGA; birth weight, 2385 +/- 69 g; relative birth weight, -2.2 +/- 0.08 SD; P < 0.001 vs. AGA newborn infants). Leptin concentrations were higher in large for gestational age (35.7 +/- 8.0 micrograms/L; P < 0.005), but lower in SGA (3.3 +/- 0.5 micrograms/L; P < 0.001) than in AGA infants (14.5 +/- 2.8 micrograms/L). When adjusted for differences in body weight, mean leptin levels were similar in the three newborn groups. Leptin concentration correlated closely with both absolute and relative birth weights (r = 0.71; P < 0.001 in both), with cord blood insulin concentration (r = 0.67; P < 0.001), and with placental weight (r = 0.60; P < 0.001). These data suggest that leptin is synthesized in utero, and that the circulating leptin concentration relates to the intrauterine growth pattern. PMID- 9329362 TI - Prophylactic adrenalectomy of a three-year-old girl with congenital adrenal hyperplasia: pre- and postoperative studies. AB - Long term follow-up studies of children with congenital adrenal hyperplasia have documented less than desirable outcomes, including reduction in final adult height, obesity, virilism, and decreased fertility. We have proposed that children with the most severe forms of congenital adrenal hyperplasia would be better off if their adrenals were removed at an early age. We report here on our experience with prophylactic bilateral adrenalectomy in a 3-yr-old girl with a double null mutation of the CYP21 gene. The results of sodium balance studies, performed preoperatively on our patient and her unaffected fraternal twin sister, and hormonal data are presented as well. In contrast to her twin, who markedly increased her sodium retention in response to ACTH, our patient showed increased natriuresis, suggesting a deleterious effect of her adrenals on sodium homeostasis. Adrenalectomy was carried out at the time of necessary genital repair. No surgical or postsurgical complications were encountered. PMID- 9329364 TI - Increased expression of the Na+/I- symporter in cultured human thyroid cells exposed to thyrotropin and in Graves' thyroid tissue. AB - The Na+/I- symporter (NIS) is important in hormone synthesis in the thyroid gland. NIS activity, as reflected by I- uptake, was increased by TSH (1 mU/mL) or forskolin (10 mumol/L) in primary cultured human thyroid cells. Northern blot analysis revealed that incubation of these cells with TSH or forskolin for 24 h increased the abundance of NIS messenger ribonucleic acid (mRNA) 2.3- and 2.5 fold, respectively. Immunoblot analysis revealed 2.7- and 2.4-fold increases, respectively, in the amount of NIS protein after 48 h, suggesting that elevated levels of intracellular cAMP induced the expression of NIS in human thyrocytes. We then studied the levels of NIS mRNA and protein in Graves' thyroid tissue and found that the amount of NIS mRNA in thyroid tissue from individuals with Graves' disease (n = 5) was 3.8 times that in normal thyroid tissue (n = 5). The abundance of NIS mRNA was significantly correlated with that of thyroid peroxidase or thyroglobulin mRNAs, but not with that of TSH receptor mRNA, in the Graves' and normal thyroid tissue specimens. The amount of NIS protein was also increased 3.1-fold in Graves' thyroid tissue compared with that in normal thyroid tissue. The increased expression of NIS may thus contribute to the development of Graves' disease. PMID- 9329365 TI - Selective growth hormone/placental lactogen gene transcription and hormone production in pre- and postmenopausal human ovaries. AB - In addition to effects of pituitary-derived gonadotropins, human GH modulates and regulates intraovarian reproductive processes in a dose-dependent manner via the endocrine GHRH/GH/insulin-like-growth-factor I (IGF-I) axis. Based on increasing evidence that ovarian regulation involves a complex system of putative para/autocrine factors, we investigated the possibility of gene-selective intraovarian GH/placental lactogen (PL) hormone production, with emphasis on differences between pre- and postmenopause. Analysis of both premenopausal (n = 8) and postmenopausal (n = 10) ovarian-derived messenger ribonucleic acid by reverse transcription-PCR, which amplifies all major gene products of the five member GH/PL gene cluster GH-N, GH-V, PL-A/B, and PL-L, revealed specific transcripts in all specimens. Their share in gene selective expression by analytical restriction enzyme digestion was determined. The expression pattern of GH/PL messenger ribonucleic acid shows PL-A/B > GH-N, which sets it apart from those of pituitary and placenta. Local production of the respective protein hormones was verified by two time-resolved immunofluorometric assays for human PL A/B and GH-N; significant amounts of these hormones were detected in cytosolic extracts of premenopausal (n = 6; 555.5 +/- 171 ng PL-A/B and 0.8 +/- 0.6 ng GH N/g tissue wet wt) and postmenopausal (n = 6; 5.2 +/- 2.7 ng PL-A/B and 0.9 +/- 0.6 ng GH-N/g tissue wet wt) ovaries. No difference was observed between pre- and postmenopausal ovarian GH-N contents, but PL values were 2-3 orders of magnitude lower in postmenopausal tissue (P < 0.001). Serum levels of healthy premenopausal (n = 21) and postmenopausal (n = 16) women were less than 0.02 ng PL/mL. In summary, ovarian-derived GH-N and PL-A/B synthesis correlates well with the established local cascade of GHRH, GHRH receptor, GH receptor, IGF-I, and IGF-I receptor as a putative para/autocrine regulator of ovarian reproductive function. PMID- 9329366 TI - Responses of catecholestrogen metabolism to acute graded exercise in normal menstruating women before and after training. AB - It has been hypothesized that exercise-related hypo-estrogenemia occurs as a consequence of increased competition of catecholestrogens (CE) for catechol-O methyltransferase (COMT). This may result in higher norepinephrine (NE) concentrations, which could interfere with normal gonadotropin pulsatility. The present study investigates the effects of training on CE responses to acute exercise stress. Nine untrained eumenorrheic women (mean percentage of body fat +/-SD: 24.8 +/- 3.1%) volunteered for an intensive 5-day training program. Resting, submaximal, and maximal (tmax) exercise plasma CE, estrogen, and catecholamine responses were determined pre- and post training in both the follicular (FPh) and luteal phase (LPh). Acute exercise stress increased total primary estrogens (E) but had little effect on total 2-hydroxyestrogens (2-OHE) and 2-hydroxyestrogen-monomethylethers (2-MeOE) (= O-methylated CE after competition for catechol-O-methyltransferase). This pattern was not significantly changed by training. However, posttraining LPh mean (+/-SE) plasma E, 2-OHE, and 2-MeOE concentrations were significantly lower (P < 0.05) at each exercise intensity (for 2-OHE: 332 +/- 47 vs. 422 +/- 57 pg/mL at tmax; for 2-MeOE: 317 +/ 26 vs. 354 +/- 34 pg/mL at tmax). Training produced opposite effects on 2-OHE:E ratios (an estimation of CE formation) during acute exercise in the FPh (reduction) and LPh (increase). The 2-MeOE:2-OHE ratio (an estimation of CE activity) showed significantly higher values at tmax in both menstrual phases after training (FPh: +11%; LPh: +23%; P < 0.05). After training, NE values were significantly higher (P < 0.05). The major findings of this study were that: training lowers absolute concentrations of plasma estrogens and CE; the acute exercise challenge altered plasma estrogens but had little effect on CE; estimation of the formation and activity of CE suggests that formation and O methylation of CE proportionately increases. These findings may be of importance for NE-mediated effects on gonadotropin release. PMID- 9329367 TI - Using dilution techniques and multifrequency bioelectrical impedance to assess both total body water and extracellular water at baseline and during recombinant human growth hormone (GH) treatment in GH-deficient adults. AB - Due to the use of various, and mostly indirect, methods to estimate total body water (TBW) and extracellular water (ECW), there is no agreement about whether body water distribution, i.e. the ECW to TBW ratio, is normal in GH-deficient (GHD) subjects at baseline and during recombinant human GH (rhGH) treatment. We studied body water distribution in 14 patients with adult-onset GHD and in 28 healthy controls. We also investigated the effect of GH replacement therapy for 4 and 52 weeks on body water distribution. All patients started with a dose of 0.6 IU rhGH/day for the first 4 weeks. After 52 weeks, the dose varied between 0.6 1.8 IU/day. TBW and ECW were measured by dilution of deuterium and bromide, respectively. Both parameters were also estimated using multifrequency bioelectrical impedance (BIA). Patients with GHD had significantly lower ECW and TBW than healthy controls. In addition, the ECW to TBW ratio was significantly lower in GHD patients than in healthy controls. Four weeks of GH treatment significantly increased body weight, TBW, ECW, and ECW/TBW. A further increase in TBW, but not ECW, was found after 52 weeks of treatment. The mean increases in TBW and ECW from the baselines were 2.5 +/- 0.3 and 2.0 +/- 0.3 L, respectively. The correlation coefficient and the estimated reliability between measured and estimated TBW and ECW at any time point were all high (> 0.91 and > 0.95, respectively). In general, both ECW and TBW were overestimated by multifrequency BIA in GHD adults. During treatment, the overestimation of both ECW and TBW diminished. The estimation error was correlated with the level of the body water compartment and the ratio of ECW to TBW. The estimated change in ECW with rhGH treatment was underestimated by multifrequency BIA. We conclude that GHD adults have lower ECW and TBW and a lower ECW to TBW ratio, as measured by dilution techniques. The ECW to TBW ratio can be normalized within 4 weeks of rhGH treatment at a dose of 0.6 IU/day. Finally, we conclude that multifrequency impedance measurements do not give valid estimates of body water compartments in the follow-up of patients with GHD. PMID- 9329369 TI - Detection of major T cell epitopes on human thyroid stimulating hormone receptor by overriding immune heterogeneity in patients with Graves' disease. AB - To examine the major immunogenic regions of the human TSH receptor (hTSHR), we examined 14 patients with Graves' disease and 14 healthy control subjects for their peripheral blood T cell proliferative responses to 29 synthetic peptides representing the entire ectodomain of the hTSHR (TSHR-ecd). By combining an analytical approach encompassing the grading of peptide-induced responses and nonparametric testing, we obtained evidence for highly significant differences (P = < 0.000001) in the patient group compared with minor differences in the control group (P = 0.045). To account for this difference, we identified four major T cell epitopes (amino acid 247-266, 202-221, 142-161, and 52-71), by multiple comparison analysis, in the patient group. Furthermore, we demonstrated by radiolabeled PCR that the responding T cells were clonally expanding. These findings demonstrate that despite likely differences in human leukocyte antigen type among patients with Graves' disease, several distinct hTSHR epitopes elicited significant responses in the immune system of patients with Graves' disease, and that such patients are most often poorly tolerant to particular epitopes of the TSH receptor ectodomain, The data support the notion of TSHR peptide antigens overriding human immune heterogeneity in patients with Graves' disease, and raise the possibility of applying analog peptide blockade to suppress T cell responsivity. PMID- 9329368 TI - Functioning thoracic paraganglioma: association with Von Hippel-Lindau syndrome. AB - Functioning thoracic paraganglioma (pheochromocytoma) is unusual and therefore suggestive of a pathogenesis distinct from that of sporadic adrenal pheochromocytoma. To determine whether the pheochromocytoma-associated syndromes Von Hippel-Lindau disease (VHL) and multiple endocrine neoplasia type 2 (MEN 2) play a role in the development of thoracic functioning paragangliomas, germline DNA from five unselected patients with this rare tumor was analyzed for mutations in the genes that predispose to VHL and MEN 2. Genetic investigations and further clinical data revealed that three had VHL, with two different germline mutations of the vhl gene, but no individual was affected by MEN 2. Two of the three patients with VHL did not show any additional VHL-associated lesions. This result suggests that VHL should be considered in the differential diagnosis of thoracic pheochromocytoma, as such a diagnosis carries further important implications for the patient and family. Conversely, in patients suspected of a catecholamine secreting tumor and known VHL, thoracic localization should be considered if an adrenal pheochromocytoma cannot be detected. PMID- 9329370 TI - Suppression of endogenous testosterone in young men increases serum levels of high density lipoprotein subclass lipoprotein A-I and lipoprotein(a). AB - We investigated the effect of testosterone suppression on lipoprotein metabolism in men. After a baseline period of 14 days, 12 healthy young men received over a period of 3 weeks daily s.c. injections of Cetrorelix, an antagonist of GnRH. The volunteers were then followed-up for 10 additional weeks. Administration of Cetrorelix suppressed testosterone significantly up to day 35, after which values returned to baseline. Suppression of testosterone was associated with significant and consistent increases in mean serum levels of high density lipoprotein (HDL) cholesterol by 20% (P < 0.0001), apolipoprotein A-I (apoA-I) by 10% (P = 0.0032), apoA-II by 7% (P = 0.0112), HDL subclass lipoprotein A-I (LpA-I) by 23% (P = 0.002), and plasma lecithin:cholesterol acyltransferase by 7% (P < 0.001). Serum levels of HDL subclass LpA-I/LpA-II changed insignificantly. Moreover, suppression of testosterone significantly increased the median of lipoprotein(a) [Lp(a)] levels from 5.5 to 8.5 mg/dL (P < 0.0001). The increase in Lp(a) levels was positively correlated with baseline levels of Lp(a) (r = 0.91; P < 0.001) and amounted to 40-60% in individuals with baseline levels of Lp(a) higher than 3 mg/dL. We conclude that endogenous testosterone is involved in the regulation of HDL cholesterol and Lp(a) levels and may thereby influence cardiovascular risk. PMID- 9329371 TI - Paraoxonase-2 gene (PON2) G148 variant associated with elevated fasting plasma glucose in noninsulin-dependent diabetes mellitus. AB - Defining the genetic determinants of NIDDM requires evidence from several complementary approaches, including both linkage and association analyses using both discrete phenotypes and intermediate quantitative traits. We tested for association between common genomic variation in three genes that map to chromosome 7q21-q22 and quantitative traits related to NIDDM in a sample of Oji Cree. We found that a common genomic variation in codon 148 (alanine or glycine) of the paraoxonase-2 gene (PON2) demonstrated a significant association with a variation in fasting plasma glucose (P < 0.0001). Furthermore, we found a significant association between a variation in fasting plasma glucose and the interaction term comprised of a PON2 codon 148 genetic variation and the presence of noninsulin-dependent diabetes mellitus (NIDDM; P < 0.0001). We then analyzed subjects according to PON2 genotype and NIDDM status. In subjects with NIDDM, the PON2 codon 148 G/G homozygotes had significantly higher mean fasting plasma glucose than subjects with the other two genotypes (P < 0.0001). However, in non NIDDM subjects, there was no difference in mean fasting plasma glucose among any of the genotypes. There was no association of the PON2 genotype with NIDDM itself, with impaired glucose tolerance, or with other quantitative traits related to NIDDM in this sample. These findings suggest that 1) the PON2 G148 gene variant worsens glycemia in subjects with NIDDM; 2) defining the physiological role of the PON2 gene product would be worthwhile; and 3) genetic factors can modify the severity of clinical phenotypes in subjects with NIDDM. PMID- 9329372 TI - A case of metastatic medullary thyroid carcinoma: early identification before surgery of an RET proto-oncogene somatic mutation in fine-needle aspirate specimens. AB - Medullary thyroid carcinoma (MTC) management requires determination of the sporadic or familial nature of the disease. RET proto-oncogene mutation analysis in the tumor tissue obtained at surgery and in the peripheral blood identifies somatic vs. germinal mutations. We now report a case of MTC in which a RET somatic mutation at codon 918 was detected in fine-needle aspiration specimens obtained from both the thyroid nodule and two enlarged neck lymph nodes but not in peripheral blood. Therefore, a diagnosis of sporadic MTC was made before surgery. Thus, this approach, by excluding preoperatively multiple endocrine neoplasia disease, permitted immediate thyroidectomy without search for pheochromocytoma. PCR-based genetic analysis in fine-needle aspiration biopsy specimens, therefore, preoperatively identifies genetic abnormalities at an early and easily manageable stage and may well contribute to the management strategy of MTC. PMID- 9329373 TI - Expression, localization, and thyrotropin regulation of cathepsin D in human thyroid tissues. AB - Enzymatic activity and isoform expression of cathepsin D (cath D) were studied in 107 cytosols from various human thyroid tissues including 21 normal tissues, 12 cold benign nodules, 17 toxic adenomas, 22 samples from Graves' disease patients, and 35 thyroid carcinomas. Cath D assay was optimized for human thyroid tissues. We found that mean cath D specific activities, expressed as units per milligrams protein minus thyroglobulin, were higher in carcinomas (P = 0.0001), toxic adenomas (P = 0.0001), and specimens from Graves' disease patients (P = 0.0001) than in normal thyroid tissues. Mean cath D activity in carcinomas was also significantly different from that in cold benign nodules (P < 0.001) and Graves' disease tissues (P < 0.05) but not from that of toxic adenomas. To determine possible mechanisms by which the observed increase in cath D activity might be regulated, we used Western blotting to measure relative amounts of cath D isoforms in the various thyroid tissues. We found that the 31-kDa major processing form of cath D was significantly increased in carcinomas and toxic adenomas compared with normal tissues (P < 0.01), cold benign nodules (P < 0.05), and Graves' disease tissues (P < 0.05). A positive correlation of cath D activity with relative expression of the 31-kDa form (r = 0.67, P = 0.0001) was observed in 104 thyroid cytosols. These data demonstrate a deregulation at the protein level, with resulting increases in cath D activity. Immunogold labeling of cath D showed particle concentration in lysosomes or phagosomes in both normal follicles and papillary carcinoma cells, indicating that cath D localization was not altered by malignant transformation in human thyroid cells. TSH induced cath D synthesis and secretion in extracellular fluid of normal human thyroid cells in primary culture; TSH had little effect on intracellular cath D level. In conclusion, TSH-induced cath D synthesis may explain high cath D levels in Graves' disease tissues and toxic adenomas, because these tissues possess a permanently stimulated cAMP transduction pathway. Furthermore, the overexpression of cath D in thyroid carcinomas in comparison with normal controls adds further arguments for the potential role of cath D in tumor growth and metastasis. PMID- 9329374 TI - Altered composition of high density lipoproteins in women with the polycystic ovary syndrome. AB - Women with polycystic ovary syndrome (PCOS) appear at increased cardiovascular risk due in part to a dyslipidemia characterized by increased plasma triglyceride and reduced high density lipoprotein (HDL) cholesterol levels. This is a detailed exploratory study of HDL composition in 35 obese [body mass index (BMI), > 27] and 22 nonobese subjects with PCOS and in 14 healthy obese and 18 nonobese women. Although we found reduced levels of total and HDL2 cholesterol in obese women with PCOS, HDL composition was modified by depletion of lipid relative to protein, with reduced ratios of HDL total cholesterol and HDL phospholipids to apolipoprotein A-I (apoA-I) compared to those in obese controls (P = 0.008 and P = 0.012, respectively). This was explained by reduced cholesterol (P = 0.004) and phospholipid (although not significant, P = 0.07) in HDL with no change in the content of apoA-I, its major protein. Obesity, insulin resistance, and hyperandrogenemia are features of PCOS and potentially affect lipid metabolism. Insulin sensitivity was assessed by the reduction in endogenous glucose concentration after exogenous insulin; the insulin, glucose, and fatty acid responses to oral glucose; and the fasting insulin concentration. When age, BMI, free androgen index, insulin sensitivity determined by all methods, and the presence of PCOS were subjected to stepwise multivariate regression analysis, the presence of PCOS was the most consistent predictor of lipid-depleted HDL (HDL total cholesterol/apoA-I and HDL phospholipids/apoA-I). We speculate that altered activity of hepatic lipase or lipid transfer protein could explain this aspect of the dyslipidemia. Obesity has an important influence on the lipid profile. Obese PCOS and control subjects had higher levels of cholesterol, triglyceride, apoB, and fatty acids than their lean counterparts, and BMI proved the best predictor of blood levels on multiple regression analysis. In contrast, lean PCOS patients had normal sensitivity to insulin and lipid profiles similar to those of the lean controls and did not manifest the HDL abnormalities. Although in PCOS, correlations were obtained between the free androgen index and cholesterol, triglyceride, and apoB levels and between the integrated glucose and insulin responses after oral glucose and fasting fatty acid and triglyceride levels, when age and adiposity were included as covariates only fatty acids and the integrated glucose response remained significantly correlated. Among the controls, total, low density lipoprotein cholesterol, triglycerides, and apoB were related to aspects of insulin sensitivity independent of age and BMI. Lipid metabolism in PCOS is dependent on several related factors, but subjects with PCOS who are obese show a specific reduction in HDL lipid, suggesting a reduced capacity for cholesterol removal from tissues with diminished antiatherogenic potential. Efforts should be directed toward reducing obesity in PCOS to improve the metabolic disturbance in addition to ameliorating the presenting symptoms. PMID- 9329375 TI - Molecular scanning of beta-3-adrenergic receptor gene in total congenital lipoatrophic diabetes mellitus. AB - Total congenital lipoatrophic diabetes is characterized by absence of subcutaneous adipose tissue, hypertriglyceridemia, and insulin resistance. We hypothesized that mutations in the beta-3-adrenergic receptor (beta 3AR) gene might result in the lipoatrophic phenotype by preventing triglyceride storage in adipocytes; thereby, resulting in secondary insulin resistance. We screened the beta 3AR gene in 7 subjects with total congenital lipoatropic diabetes. We found a heterozygous substitution of a guanine to cytosine at position -153 (G-153C) in the 5'-untranslated region of 3 African-American lipoatrophic siblings and 1 sibling without lipoatrophy but with insulin resistance. To determine whether the base change was related to the lipoatrophic phenotype, we genotyped 69 African Americans without lipoatrophy and found the G-153C substitution in 2 control subjects (allele frequency = 0.01). No other single-stranded polymorphism variants were found in any of the 7 lipoatrophic subjects. Direct sequencing of both alleles of 1 lipoatrophic subject demonstrated a thymidine insertion at position -300 in both alleles. All lipoatrophic subjects along with 20 African American control subjects were homozygous for the base insertion, suggesting an error in the published sequence. In conclusion, mutations in the beta 3AR gene do not appear to be involved in the development of congenital total lipoatrophy. PMID- 9329376 TI - Transport and metabolism of thyrotrophin-releasing hormone across the fetal membrane. AB - To determine the transfer and metabolism of TRH by human fetal membranes, the bidirectional transport and uptake of TRH was investigated by adding 125I-labeled TRH (100,000 cpm) or commercial TRH either to the maternal or the fetal compartment of an in vitro model of cultured human fetal membranes obtained from term and preterm placenta. Transmembrane transfer was also studied in the presence of 200 microM p-hydroxy-mercuriphenyl-sulphonic acid (p-HMSA), a dipeptidase enzyme inhibitor. Creatinine and heparin were used as an internal markers. Metabolites of TRH were separated from intact molecules by gel filtration on Sephadex G-10. The structural integrity of the membrane was confirmed by electron microscopy. The transmembrane transfer of radiolabeled and commercial TRH were comparable across both preterm and term placenta. When transport was studied from the maternal to fetal side, the maternal concentration of TRH declined rapidly from 100% at time 0 to 19.31 +/- 2.26% at 8 h with a concomitant increase in the fetal concentration from undetectable to a maximum of 2.56 +/- 0.38% with a fetomaternal ratio of 0.16 +/- 0.01. Transfer of TRH from the fetal to maternal compartment was similar to that of maternal to fetal. Chromatography of maternal and fetal media showed that TRH was metabolized by the membrane into small molecular weight fragments. Treatment of the membrane with p HMSA increased TRH transport from the maternal to fetal compartment to 18.12 +/- 0.91 (P < 0.001) with an fetomaternal ratio of 0.35 +/- 0.02 (P < 0.001). Although transmembrane transfer of TRH from the fetal to maternal side was also increased by p-HMSA, levels achieved were less than that from maternal to fetal (12.26 +/- 1.50%; P < 0.05). These results suggest that the human fetal membrane acts as an enzymatic barrier to the bidirectional transfer of TRH from 24 weeks gestation. PMID- 9329377 TI - Leptin concentrations in relation to body mass index and the tumor necrosis factor-alpha system in humans. AB - The expression of leptin, an adipocyte-derived protein whose circulating levels reflect energy stores, can be induced by tumor necrosis factor (TNF)alpha in rodents, but an association between the TNF alpha system and leptin levels has not been reported in humans. To evaluate the potential association between serum leptin and the TNF alpha system, we measured the levels of soluble TNF alpha receptor (sTNF alpha-R55), which has been validated as a sensitive indicator of activation of the TNF alpha system. We studied two groups: 1) 82 young healthy normal controls and 2) 48 patients with noninsulin dependent diabetes mellitus (NIDDM) and 24 appropriately matched controls. By simple regression analysis in controls, there was a strong positive association between leptin and 3 parameters: body mass index, sTNF alpha-R55, and insulin levels. In a multiple regression analysis model, leptin remained significantly and strongly associated with body mass index, and the association of leptin with both insulin and sTNF alpha-R55, although weakened, remained significant. Patients with NIDDM had leptin concentrations similar to controls of similar weight. Importantly, serum levels of sTNF alpha-R55 were also positively and independently associated with leptin in this group of diabetic subjects and matched controls. These data are consistent with the hypothesis that the TNF alpha system plays a role in regulating leptin levels in humans. Further elucidation of a possible role of the TNF alpha system in leptin expression and circulating levels may have important implications for our understanding of obesity and cachexia in humans. PMID- 9329378 TI - Estrogen and testosterone, but not a nonaromatizable androgen, direct network integration of the hypothalamo-somatotrope (growth hormone)-insulin-like growth factor I axis in the human: evidence from pubertal pathophysiology and sex steroid hormone replacement. AB - Activation of the gonadotropic and somatotropic axes in puberty is marked by striking amplification of pulsatile neurohormone secretion. In addition, each axis, as a whole, constitutes a regulated network whose feedback relationships are likely to manifest important changes at the time of puberty. Here, we use the regularity statistic, approximate entropy (ApEn), to assess feedback activity within the somatotropic (hypothalamo-pituitary/GH-insulin-like growth factor I) axis indirectly. To this end, we studied pubertal boys and prepubertal girls or boys with sex-steroid hormone deficiency treated short-term with estrogen, testosterone, or a nonaromatizable androgen in a total of 3 paradigms. First, our cross-sectional analysis of 53 boys at various stages of puberty or young adulthood revealed that mean ApEn, taken as a measure of feedback complexity, of 24-h serum GH concentration profiles is maximal in pre- and mid-late puberty, followed by a significant decline in postpubertal adolescence and young adulthood (P = 0.0008 by ANOVA). This indicates that marked disorderliness of the GH release process occurs in mid-late puberty at or near the time of peak growth velocity, with a return to maximal orderliness thereafter at reproductive maturity. Second, oral administration of ethinyl estradiol for 5 weeks to 7 prepubertal girls with Turner's syndrome also augmented ApEn significantly (P = 0.018), thus showing that estrogen per se can induce greater irregularity of GH secretion. Third, in 5 boys with constitutionally delayed puberty, im testosterone administration also significantly increased ApEn of 24-h GH time series (P = 0.0045). In counterpoint, 5 alpha-dihydrotestosterone, a nonaromatizable androgen, failed to produce a significant ApEn increase (P > 0.43). We conclude from these three distinct experimental contexts that aromatization of testosterone to estrogen in boys, or estrogen itself in girls, is likely the proximate sex-steroid stimulus amplifying secretory activity of the GH axis in puberty. In addition, based on inferences derived from mathematical models that mechanistically link increased disorderliness (higher ApEn) to network changes, we suggest that sex-steroid hormones in normal puberty modulate feedback within, and hence network function of, the hypothalamo-pituitary/GH insulin-like growth factor I axis. PMID- 9329379 TI - Both hypothyroidism and hyperthyroidism enhance low density lipoprotein oxidation. AB - Hypothyroidism is frequently associated with hypercholesterolemia and an increased risk for atherosclerosis, whereas hyperthyroidism is known to precipitate angina or myocardial infarction in patients with underlying coronary heart disease. We have shown previously that L-T4 functions as an antioxidant in vitro and inhibits low density lipoprotein (LDL) oxidation in a dose-dependent fashion. The present study was designed to evaluate the changes in LDL oxidation in subjects with hypothyroidism and hyperthyroidism. Fasting blood samples for LDL oxidation analyses, lipoprotein determinations, and thyroid function tests were collected at baseline and after the patients were rendered euthyroid. The lag phase (mean +/- SEM hours) of the Cu+2-catalyzed LDL oxidation in the hypothyroid state and the subsequent euthyroid states were 4 +/- 0.0.65 and 14 +/ 0.68 h, respectively (P < 0.05). The lag phase during the hyperthyroid phase was 6 +/- 0.55 h, and that during the euthyroid phase was 12 +/- 0.66 h (P < 0.05). The total and LDL cholesterol levels were higher in hypothyroidism than in euthyroidism and were lower in hyperthyroidism than in the euthyroid state. We conclude that LDL has more susceptibility to oxidation in both the hypothyroid and hyperthyroid states. Thus, the enhanced LDL oxidation may play a role in the cardiac disease process in both hypothyroidism and hyperthyroidism. PMID- 9329380 TI - Raloxifene and estrogen: comparative bone-remodeling kinetics. AB - The pattern of changes in human bone remodeling produced by raloxifene (60 mg/day) was compared to that of estrogen (given as hormone replacement therapy) in 33 early postmenopausal women randomly assigned to raloxifene, estrogen, or no treatment. Remodeling was measured using calcium tracer kinetic methods employed under a constant diet and full metabolic balance conditions. Studies were performed at baseline and, to detect both early and late remodeling changes, at 4 and 31 weeks of treatment. Both raloxifene and estrogen produced a significant positive calcium balance shift at each treatment measurement point: +74 and +60 mg/day at 4 weeks, and +60 and +91 mg/day at 31 weeks for raloxifene and estrogen, respectively. Externally, this balance change was due to a highly significant fall in the urinary calcium level and marginal improvement in calcium absorption efficiency. Internally, bone resorption was significantly reduced at both measurement points: -64 and -60 mg/day at 4 weeks, and -82 and -162 mg/day at 31 weeks for raloxifene and estrogen, respectively. Bone formation was not significantly affected by either agent at 4 weeks; at 31 weeks, formation was reduced by estrogen, but not by raloxifene. Thus, at 4 weeks, the general pattern of remodeling change was identical for the two agents. At 31 weeks, remodeling suppression was greater for estrogen than for raloxifene; however, remodeling balance was the same for the two agents. We conclude that raloxifene and estrogen affect the bone remodeling apparatus similarly, and that raloxifene, therefore, is acting on bone as an estrogen agonist. PMID- 9329381 TI - Prevalence of goiter and urinary iodine excretion levels in children around Chernobyl. AB - The prevalence of goiter among children living in areas affected by the Chernobyl accident was investigated by analysis of data on approximately 120,000 children examined at five medical diagnostic centers in Belarus, Russia, and the Ukraine. Examinations of thyroid gland were conducted with an arch-automatic ultrasonographic instrument at the five centers under the same protocol. The diagnosis of goiter was established when the thyroid volume exceeded a limit calculated from age, height, and body weight of a child. A considerable variation by region was noted in the prevalence of goiter. Highest in the Kiev region, the prevalence in the five regions was 54% in Kiev, 38% in the Zhitomir regions of the Ukraine, 18% in Gomel, 22% in the Mogilev regions of Belarus, and 41% in the Bryansk region of Russia. Urinary iodine content was measured in approximately 5700 children, and an endemic iodine deficient zone was confirmed in the Bryansk, Kiev, and Zhitomir regions. A significant negative correlation was observed between the prevalence of goiter and the median level of urinary iodine content (Spearman's rank correlation coefficient was -0.35, P = 0.025). PMID- 9329382 TI - Detection of skewed X-inactivation in two female carriers of vasopressin type 2 receptor gene mutation. AB - Most cases of congenital nephrogenic diabetes insipidus (NDI) are inherited in an X-linked manner, which is due to the mutations of the vasopressin type 2 receptor (V2R) gene. However, recent reports have presented female NDI patients with heterozygote V2R gene mutations. The mechanism of inheritance was thought to be skewed X-inactivation. We present a family with congenital NDI. Three male members were diagnosed with NDI, and examination of their V2R gene revealed a G inserted at nucleotide 804 of the open reading frame. Three female individuals display different degrees of symptoms of NDI, and all of them possess both the normal and abnormal genes. The X-inactivation patterns of the female members were investigated via the detection of methylated trinucleotide repeat in the human androgen receptor gene. The grandmother showed extremely skewed methylation of one X chromosome, and the mother revealed moderately skewed methylation. The daughter of the grandmother's sister, who has no symptoms of NDI, showed random methylation. The highly skewed X-inactivation pattern of the grandmother suggests that her NDI phenotype is caused by dominant methylation of the normal allele of V2R gene. PMID- 9329383 TI - Genetic exclusion of 14 candidate genes in lipoatropic diabetes using linkage analysis in 10 consanguineous families. AB - Lipoatropic diabetes (LD) is a rare recessive autosomal disorder, mainly characterized by lipoatrophy with alterations in lipid metabolism and extreme insulin resistance. To identify molecular defects responsible for this disease, we tested the implication of 14 candidate genes coding for proteins involved either in insulin action, i.e. insulin receptor, insulin receptor substrate 1, insulin-like growth factor I receptor, diabetes-associated ras-like protein (Rad), and glycogen synthase, or in lipid metabolism, i.e. lipoprotein lipase; apolipoproteins CII, AII, and CIII; hepatic lipase; hormone-sensitive lipase; the beta 3-adrenergic receptor; leptin; and fatty acid-binding protein 2. To this end, haplotype and linkage analyses using genotyping with microsatellites in 10 consanguineous families provided us with powerful genetic tools. Our results show that in most families, lod scores at a null recombination fraction were less than -2. Haplotype analysis also argues against the involvement of these genes in LD. This implies that mutations in these genes are unlikely to make a major genetic contribution to LD. PMID- 9329384 TI - Endothelin-1 stimulates steroid secretion of human adrenocortical cells ex vivo via both ETA and ETB receptor subtypes. AB - The role played by endothelins (ETs) and their receptor subtypes (ETA and ETB) in the regulation of steroid hormone secretion in human adrenal gland remains unclear. Therefore, we investigated the gene expression of ET-1 and its receptors in highly pure preparations of human adrenocortical cells and the effect of ET-1 on their secretory activity. Reverse transcription-PCR with primers specific for prepro-ET-1, ET-converting enzyme-1, ETA, and ETB complementary DNAs demonstrated the expression of all of these genes in human adrenocortical cells. ET-1 increased the secretion of aldosterone and cortisol by enhancing both earlier and late steps of their synthesis. The secretory response to ET-1 was partially (60%) inhibited by BQ-123 and BQ-788, which are selective antagonists of the ETA and ETB receptors, respectively. When added together, the two antagonists suppressed the secretagogue effect of ET-1. Collectively, these findings suggest that ET-1, acting via both ETA and ETB receptors, may exert an autocrine/paracrine regulation of the function of the human adrenal cortex. PMID- 9329385 TI - Pituitary stalk interruption syndrome: a clinical-biological-genetic assessment of its pathogenesis. AB - The detection of pituitary stalk interruption syndrome (PSIS) by magnetic resonance imaging is a diagnostic marker of permanent GH deficiency (GHD), but the pathogenesis of PSIS is unknown. Fifty-one patients (27 males) with GHD and PSIS were classified according to whether the GHD was isolated (group 1, 16 cases) or associated with other anterior pituitary abnormalities (group 2, 35 cases). The 2 groups had similar characteristics (frequencies of perinatal abnormalities, ages at occurrence of first signs and at diagnosis, height, GH peak response to stimuli other than GHRH), but associated malformations were less frequent in group 1 (12%) than in group 2 (54%; P < 0.01), hypoglycemia occurred in 25% of group 1 and 70% of group 2 (P < 0.01), and the GH peak response to GHRH was less than 10 micrograms/L in 0% of group 1 (4 cases evaluated) and 57% of group 2 (21 cases; P < 0.05). Thirty-one cases (61%; 25 from group 2) had features suggesting an antenatal origin: familial form (4 cases), microphallus (10 boys), and/or associated malformations (50%; 21 cases). Twenty-seven cases (53%, 22 from group 2) had features suggesting a hypothalamic origin. The three group 1 patients with a GH peak of 1 microgram/L or less had no large GH-N gene deletion. One familial form had no linkage between the GHD phenotype and abnormal GH-N locus, and the only mutation described to date in the GHRH receptor gene was absent. The two patients with low plasma PRL levels had no Pit-1 gene abnormality. Thus, most of the patients with GHD associated with multiple anterior pituitary abnormalities and PSIS have features suggesting an antenatal origin. The GH-N, GHRH receptor, and Pit-1 genes do not seem to be implicated in PSIS. PMID- 9329386 TI - Oral administration of growth hormone (GH) releasing peptide-mimetic MK-677 stimulates the GH/insulin-like growth factor-I axis in selected GH-deficient adults. AB - To determine the effect of the GH releasing peptide (GHRP)-mimetic, MK-677, on the GH/insulin-like growth factor-I (IGF-I) axis in selected GH-deficient adults, we studied nine severely GH-deficient men [peak serum GH concentration in response to insulin-induced hypoglycemia of 1.2 +/- 1.5 micrograms/L, mean +/- SD (range 0.02-4.79)], age 17-34 yr, height 168 +/- 1.5 cm, body mass index 22.6 +/- 3.3 kg/m2, who had been treated for GH deficiency with GH during childhood. In a double-blind rising-dose design, subjects received once daily oral doses of 10 or 50 mg MK-677 or placebo for 4 days over two treatment periods separated by at least 28 days. Four subjects received placebo and 10 mg/day MK-677 in a cross over fashion in periods 1 and 2. Five subjects received 10 mg and then 50 mg/day MK-677 in a sequential, rising-dose fashion in periods 1 and 2, respectively. Blood was collected every 20 min for 24 h before treatment and at the end of each period for GH measurement using an ultrasensitive assay. The drug was generally well tolerated, with no significant changes from baseline in circulating concentrations of cortisol, PRL, and thyroid hormones. Serum IGF-i and 24-H mean GH concentrations increased in all subjects after treatment with both 10 and 50 mg/day MK-677 vs. baseline. After treatment with 10 mg MK-677, IGF-I concentrations increased 52 +/- 20% (65 +/- 6 to 99 +/- 9 micrograms/L, geometric mean +/- intrasubject SE, P < or = 0.05 vs. baseline), and 24 h mean GH concentrations increased 79 +/- 19% (0.14 +/- 0.01 to 0.26 +/- 0.02 microgram/L, P < or = 0.05 vs. baseline). Following treatment with 50 mg MK-677, IGF-I concentrations increased 79 +/- 9% (84 +/- 3 to 150 +/- 6 micrograms/L, P < or = 0.05 vs. baseline) and 24-h mean GH concentrations increased 82 +/- 29% (0.21 +/- 0.02 to 0.39 +/- 0.04 microgram/L, P < or = 0.05 vs. baseline), respectively. Serum IGF binding protein-3 concentrations increased with both 10 mg (1.2 +/- 0.1 to 1.7 +/- 0.1 micrograms/L, P < or = 0.05) and 50 mg MK-677 (1.7 +/- 0.1 to 2.2 +/- 0.2 micrograms/L, P < or = 0.05). The GH response to MK-677 was greater in subjects who were the least GH/IGF-I deficient at baseline; by linear regression analysis the increase in 24-h mean GH concentration was positively related to both baseline 24-h mean GH concentration (r = 0.81, P = 0.009) and baseline IGF-I (r = 0.79, P = 0.01) for 10 mg MK-677. IGF-I responses were not significantly related to any baseline measurement. Fasting and postprandial insulin and postprandial glucose increased significantly after MK-677 treatment, and the clinical significance of these changes will need to be assessed in longer term studies. Oral administration of such GHRP-mimetic compounds may have a role in the treatment of GH deficiency of childhood onset. PMID- 9329387 TI - Natural antiestrogen receptor autoantibodies in man with estrogenic activity in mammary carcinoma cell culture: study of their mechanism of action; evidence for involvement of estrogen-like epitopes. AB - We previously reported that human natural autoantibodies enriched in antiestrogen receptor Ig (IgGs) display estrogenic activity in MCF-7 mammary carcinoma cells. In this study, we investigated IgGs' mechanism of action. We showed that: 1) IgGs Fab fragments (which contain only one antigen binding site) induced an estrogenic response in MCF-7 cells, producing estrogen receptor (ER) down-regulation and an increase in progesterone receptor concentration; 2) IgGs specifically inhibited MCF-7 cell surface labeling with fluorescent estradiol (E2)-BSA conjugates; 3) this inhibition of E2-BSA binding to membrane estrogen binding sites was largely caused by natural anti-E2-BSA antibodies (Ab) selectively associated with the natural anti-ER Ab within IgGs; 4) furthermore, these natural anti-E2-BSA Ab accounted for most of IgGs estrogenic activity in cell culture; 5) however, when incubated with cytosolic ER, they did not behave like estrogens, but they decreased ER hormone binding capacity; and 6) although IgGs stimulated cAMP production, their anti-E2-BSA Ab subpopulation did not. In conclusion, the estrogenic activity of IgGs does not involve Ab mimicking E2 molecular configuration or ligand-independent cAMP mediated pathways, membrane Fc receptors, and membrane receptor cross-linking mechanisms. On the contrary, IgGs seem to function by neutralizing estrogen-like epitopes, associated with ER related peptides, which might inhibit ER activation. PMID- 9329388 TI - Mutations of the human thyrotropin receptor gene causing thyroid hypoplasia and persistent congenital hypothyroidism. AB - The pathogenesis of congenital hypothyroidism due to thyroid dysgenesis is still unknown. A point mutation in the TSH receptor (TSHR) of the hypothyroid hyt/hyt mouse invoked the TSHR as a candidate gene for congenital hypothyroidism. Therefore, we screened for mutations in the TSHR gene in patients with congenital hypothyroidism and hypoplasia of the gland. In one girl detected in neonatal screening with the confirmed diagnosis of permanent congenital hypothyroidism with reduced thyroid volume, two novel mutations in the TSHR gene were identified. Single strand conformational polymorphism and subsequent DNA sequencing studies of a fragment of the TSHR gene showed that the patient is a compound heterozygote for 2 loss of function mutations in exon 10 of the TSHR gene. In the mutant maternal allele, 18 nucleotides (positions 1217-1234) are deleted, and 4 novel bp are inserted, resulting in a frame-shift and premature termination of the coding sequence. Transfection studies showed that this truncated TSHR was trapped intracellularly and completely lacked cell surface expression. The paternal gene harbors a missense mutation at nucleotide position 1170, leading to the exchange of the highly conserved C-390 for a W residue. This alteration resulted in a drastic loss of affinity and potency of TSH acting at the mutant compared to the wild-type receptor. In contrast to the published loss of function mutations of the TSHR leading to euthyroid hyperthyrotropinemia, the two new mutations lead to persistent congenital hypothyroidism and defective organ development. Further studies will have to analyze to what extent TSHR mutations are involved in the pathogenesis of congenital hypothyroidism as opposed to other genetic or environmental factors. PMID- 9329389 TI - Tumor-specific decreased expression of calcium sensing receptor messenger ribonucleic acid in sporadic primary hyperparathyroidism. AB - Secretion of PTH is regulated by extracellular calcium via calcium receptors (CaR) on the parathyroid cell surface. Recent studies have shown a decreased expression of CaR messenger RNA (mRNA) and CaR protein in pathological parathyroids. We studied the expression of CaR mRNA in pairs of adenoma and adenoma-associated normal gland from the same patients (n = 17) and in biopsies of normal parathyroid glands of normocalcemic subjects (n = 4) using in situ hybridization with oligonucleotide probes on frozen sections. No down-regulation of CaR mRNA caused by hypercalcemia could be demonstrated in the normal adenoma associated parathyroids when compared with the normal parathyroids of normocalcemic subjects. In contrast, CaR mRNA in the adenomas was significantly reduced to 64% (median; range 41-98) of the corresponding normal adenoma associated glands. No correlation was seen between CaR mRNA in the adenoma and preoperative serum calcium, PTH, or weight of the adenoma. Loss of heterozygosity studies were performed on adenomas using markers for the locus of the CaR gene on chromosome 3q. No allelic loss was demonstrated, excluding allelic loss as the cause for decreased CaR mRNA expression in the adenomas. It is concluded that the lowered levels of CaR mRNA in parathyroid adenomas may contribute to the increased set point of PTH secretion. In large adenomas the increased cell mass seems to be more important for the increased secretion of PTH. PMID- 9329390 TI - A large multiple endocrine neoplasia type 1 family with clinical expression suggestive of anticipation. AB - We describe a large multigenerational multiple endocrine neoplasia Type 1 (MEN1) family with clinical expression suggestive of anticipation. In the second and third generations, two deceased obligate gene carriers died at the ages of 85 and 76 without the history of MEN1, whereas two other living gene carriers above the age of 65 have had no clinical evidence of MEN1 to date. In the fourth generation, eight members were affected, with four having severe MEN1-related and atypical malignancies: a case of metastatic endocrine pancreatic tumor, two cases of metastatic thymic carcinoids, and a case of spinal ependymoma. In the fifth generation, all five patients were below the age of 22 when the disease was detected. MEN1 was confirmed in the family by linkage analysis using MEN1-linked microsatellite markers and by identification of a nonsense mutation in the MEN1/menin gene. Alleotyping showed loss of heterozygosity (LOH) involving the wild-type alleles in seven tumors in the family including the ependymoma, which is the first MEN1-related case that shows genetic abnormality in chromosome 11q13, suggesting that MEN1 gene might be involved in the tumorigenesis of a subset of ependymomas. In relation to clinical anticipation, repeated expansion studies were carried out but failed to detect any expansion. We conclude that this is a unique MEN1 family and that an unknown genetic mechanism might be contributing to the anticipation phenomenon. We demonstrate in this family that all gene carriers, including the very young members, will need close and careful follow-up. PMID- 9329391 TI - The localization of androgen receptors in human bone. AB - Androgens have important effects on the human skeleton, and deficiency has been associated with bone loss in both males and females. The skeletal actions of androgens may be mediated directly via the androgen receptor (AR) or indirectly via the estrogen receptor after aromatization to estrogens. The presence of androgen receptors has been demonstrated in bone cells and chondrocytes in vitro, but their presence in human bone in situ has not been reported. In order to provide further evidence for a direct action of androgens on bone via androgen receptors, we have used specific monoclonal antibodies to investigate the expression of human AR in normal developing and osteophytic bone of both sexes. In the growth plates from the developing bone, androgen receptors were predominantly expressed in hypertrophic chondrocytes and in osteoblasts at sites of bone formation. They were also observed in osteocytes in the bone, and in mononuclear cells and endothelial cells of blood vessels within the bone marrow. In the osteophytes, androgen receptors were widely distributed at sites of endochondral ossification in proliferating, mature, and hypertrophic chondrocytes and at sites of bone remodeling in osteoblasts. They were also expressed in osteocytes and mononuclear cells within the bone marrow. The pattern and number of cells expressing the receptor was similar in both sexes. Our results show for the first time the presence and distribution of androgen receptors in normal developing human and osteophytic bone in situ and further provide evidence for a direct action of androgens on bone and cartilage cells. PMID- 9329392 TI - Effect of 12-month dehydroepiandrosterone replacement therapy on bone, vagina, and endometrium in postmenopausal women. AB - The effect of 12-month dehydroepiandrosterone (DHEA) replacement therapy has been evaluated in 14 60- to 70-yr-old women who received daily applications of a 10% DHEA cream. Vaginal epithelium maturation was stimulated by DHEA administration in 8 of 10 women who had a maturation value of zero at the onset of therapy, whereas a stimulatory effect was also seen in all three women who had an intermediate vaginal maturation index before therapy. The estrogenic effect of DHEA observed in the vagina was not observed in the endometrium, which remained atrophic in all women. Most interesting, the bone mineral density significantly increased at the hip from 0.744 +/- 0.021 to 0.759 +/- 0.025 g/cm2 after 12 months of treatment (P < 0.05). These changes in bone mineral density were associated with a significant 20.0% decrease (P < 0.01) in plasma bone alkaline phosphatase and a 28% decrease in the urinary hydroxyproline/creatinine ratio. A 2.1-fold increase over the control value (P < 0.01) in plasma osteocalcin was concomitantly observed. The present data describe for the first time a series of medically important beneficial effects of DHEA therapy in postmenopausal women through transformation of the precursor steroid DHEA into androgens and/or estrogens in specific peripheral intracrine tissues without significant adverse effects. The stimulatory effect on the vaginal epithelium in the absence of stimulation of the endometrium is of particular interest because it eliminates the need for progestin replacement therapy. On the other hand, the stimulatory effect on bone mineral density accompanied by an increase in serum osteocalcin, a marker of bone formation, suggests stimulation of bone formation by the androgenic action of DHEA, a finding of particular interest for both the prevention and treatment of osteoporosis. PMID- 9329393 TI - Evidence for endocrinological abnormalities in heterozygotes for adrenal 11 beta hydroxylase deficiency of a family with the R448H mutation in the CYP11B1 gene. AB - In about 5% of cases of classical congenital adrenal hyperplasia, steroid 11 beta hydroxylase deficiency is the underlying defect. In two publications, no biochemical abnormalities have been reported in obligate heterozygotes for 11 beta-hydroxylase deficiency. We found the typical plasma steroid pattern of 11 beta-hydroxylase deficiency and identified the R448H mutation in the CYP11B1 gene in a boy presenting with pseudoprecocious puberty at age 2 yr. Both parents and an older sister were genotyped and were heterozygous carriers for the R448H mutation in CYP11B1. In contrast to the data reported in the literature, we found increased responses of plasma 11-deoxycortisol and 11-deoxycorticosterone in the short term ACTH test in the three family members heterozygous for the R448H mutation. PMID- 9329394 TI - Tissue distribution of estrogen receptors alpha (ER-alpha) and beta (ER-beta) mRNA in the midgestational human fetus. AB - We compared the expression profiles of the mRNAs of both estrogen receptors, ER alpha and the recently cloned ER-beta, in the midgestational human fetus by semiquantitative RT-PCR. ER-alpha was most abundant in the uterus, and smaller quantities were detected in the ovary, testis, skin and gut. High amounts of ER beta mRNA were present in fetal ovaries, testes, adrenals and spleen. In these tissues, the levels of ER-beta mRNA were higher than ER-alpha. In the uterus, however, ER-alpha mRNA was more abundant, and ER-beta mRNA was expressed only moderately. ER-beta mRNA was present at moderate to low levels in the thymus, pituitary gland, skin, lung, kidney and brain cortex. In the course of our work, using the ER-beta primers on genomic DNA, an intron of 2468 bp in length, located between nt 222 and 223 in the A/B domain of ER-beta cDNA, was detected, cloned and sequenced. The study shows that the expression profile of the two ERs is different, and ER-beta is expressed in a variety of tissues during human fetal development, suggesting different, organ-specific roles for the two receptors. PMID- 9329395 TI - The diagnosis of growth hormone deficiency in adults. PMID- 9329396 TI - Not only growth hormone (GH)-deficient men are more responsive to GH than women. PMID- 9329397 TI - Pseudoacromegaly and hyperinsulinemia: a possibility of premature atherosclerosis? PMID- 9329399 TI - Spontaneous complete remission of primary pigmented adrenocortical disease. PMID- 9329398 TI - 1,25 Dihydroxyvitamin D and cancer. PMID- 9329400 TI - Thyroglobulin-like immunoreactivity within goitrous thyroid stroma after iodine overload. PMID- 9329401 TI - What should be considered a low dose in the ACTH stimulation test? PMID- 9329402 TI - Short bowel syndrome: remedial features that influence outcome and the duration of parenteral nutrition. PMID- 9329403 TI - Safety of heparin use in the premature infant. PMID- 9329404 TI - The stickiness of newborn skin: bioadhesion and the epidermal barrier. PMID- 9329405 TI - Rescue in inner space: management of Rh hemolytic disease. PMID- 9329406 TI - Expired nitric oxide in pediatric asthma: emissions testing for children? PMID- 9329407 TI - Evaluation of severe hypospadias. PMID- 9329408 TI - Childhood asthma and allergic rhinitis: the role of leukotrienes. AB - Research in the past two decades has shown that patients with asthma and rhinitis have inflammation of the involved tissues. This perception has been reflected in recent treatment guidelines, which stress the decreased use of symptom-based therapy and increased use of antiinflammatory therapies to control underlying inflammation. Corticosteroids are the most effective drugs currently in use; however, their use may be limited by potential problems with safety and patient/family adherence, which includes the "fear factor." In addition, the use of high doses of topical corticosteroids (especially when used in both the nose and airways) may have adverse effects when used continuously for long periods. The inflammatory response is complex, involving numerous inflammatory mediators and cells that interact in complicated and interrelated pathways. This provides researchers with numerous interactions at which molecular intervention may result in the attenuation of inflammation, and thus clinical disease. The leukotrienes, a group of important inflammatory mediators, cause vascular leakage and tissue edema; they also promote mucus secretion and a potent bronchoconstriction in patients with asthma. Currently a number of antileukotriene drugs have been developed and preliminary research indicates that they may provide clinicians with a non-steroidal antiinflammatory therapy that may provide steroid-sparing effects. This review examines the leukotrienes and the effects of antileukotriene agents in patients with asthma and allergic rhinitis. PMID- 9329409 TI - Influence of bacterial overgrowth and intestinal inflammation on duration of parenteral nutrition in children with short bowel syndrome. AB - OBJECTIVES: Massive intestinal resection results in short bowel syndrome and necessitates prolonged parenteral feeding. The purpose of this work was to assess the impact of late complications of short bowel syndrome, including intestinal bacterial overgrowth and enterocolitis, on the duration of parenteral nutrition (PN) in comparison with factors evident in the neonatal period. METHODS: Retrospective chart review. RESULTS: Of 49 children, 42 were weaned from parenteral nutrition after a treatment course of 17 +/- 14 months. In these 42, postresection small intestinal length equaled 81 +/- 65 cm; 45% had an ileocecal valve. Small intestinal length in the seven children who were PN dependent was 31 +/- 30 cm (p < 0.05); none had an ileocecal valve (p < 0.05). Bacterial overgrowth occurred in all seven PN-dependent children and in 23 of 42 children eventually weaned from PN (p < 0.05). When bacterial overgrowth was identified before weaning (n = 12), the duration pf PN was 28 +/- 17 months, but when bacterial overgrowth was first identified only after weaning (n = 11), the duration of PN was 16 +/- 13 months (p < 0.05). Small intestinal inflammation correlated with bacterial overgrowth (r = 0.69). Those children with severe enteritis identified before weaning remained on the PN regimen for 36 +/- 15 months, in comparison with 21 +/- 14 months in those with mild enteritis and 13 +/- 11 months in those without inflammation (p < 0.02). CONCLUSIONS: Although the length of small intestine remaining after resection is the best immediate predictor of final success in terminating PN in children with short bowel syndrome, PN is prolonged by bacterial overgrowth and associated enteritis in those who will ultimately be weaned. PMID- 9329410 TI - Heparin and the risk of intraventricular hemorrhage in premature infants. AB - OBJECTIVE: This study was carried out to determine whether the routine use of low dose heparin in umbilical catheter infusates increases the risk of intraventricular hemorrhage or alters the coagulation profile in premature infants. METHODS: In a randomized, blinded trial, 113 infants born at less than 31 weeks' gestation were assigned to receive, in their umbilical catheter infusate, either 1 unit of heparin per milliliter (n = 55) or no heparin (n = 58). Prothrombin time, activated partial thromboplastin time, fibrinogen concentration, and antithrombin III activity levels were determined at the start and the completion of the study. Cranial ultrasonography was performed during the first week of life. RESULTS: There was no difference in the incidence of intraventricular hemorrhage between the heparin and no heparin groups, 35.8% and 31.5%, respectively (p = 0.6). Similarly, no difference was detected in the incidence of severe intraventricular hemorrhage (grades III/IV). Prothrombin time, activated partial thromboplastin time, and fibrinogen levels were not significantly different between the two groups. However, the use of heparin was associated with a lower antithrombin III activity level. Antenatal indomethacin use was associated with a 2.9 increased risk of intraventricular hemorrhage (95% confidence interval, 1.15 to 7.17). CONCLUSION: A low dose of heparin added to umbilical catheter infusates does not increase the incidence or severity of intraventricular hemorrhage or significantly alter the coagulation profile in premature infants. PMID- 9329411 TI - Disruption of barrier function in neonatal skin associated with adhesive removal. AB - OBJECTIVE: Patients in the neonatal intensive care unit require life support and monitoring equipment that must be securely attached to the skin; removal or replacement often causes skin trauma. In this study, we compared the effects of application and removal of three different adhesives on the skin barrier function of premature neonates. The effects were measured by transepidermal water loss (TEWL), colorimetric measurements, and visual inspection. DESIGN: Thirty neonates, between 26 and 40 weeks of gestational age and with birth weights ranging from 690 to 3000 gm, were enrolled in the study during the first week of life. Pieces of plastic tape (1 cm2), pectin barrier, and hydrophilic gel were applied to previously undisturbed sites on the back. A fourth site was used as a control. We measured TEWL, colorimetric readings, and visual inspection scores of skin irritation and stripping at each of the four sites serially: before adhesive application, 30 minutes after adhesive removal, and 24 hours later. RESULTS: Thirty minutes after adhesive removal, TEWL, colorimetric measurements, and visual inspection scores were all significantly higher at the sites of plastic tape and pectin barrier removal than at the control and gel adhesive sites (p < 0.01), demonstrating greater disruption of skin barrier function with removal of the plastic tape and pectin barrier. When the neonates were divided into three groups on the basis of birth weight (< 1000 gm [n = 10], 1000 to 1500 gm [n = 11], and > 1500 gm [n = 9], the same pattern of greater disruption in skin barrier function, as measured by TEWL, was observed in each birth weight group. Twenty-four hours after adhesive removal, TEWL of the plastic tape and pectin barrier sites were not significantly different from the control site, indicating recovery of skin barrier function. CONCLUSIONS: This study demonstrates that a single application and removal of two commonly used adhesives, plastic tape and pectin barrier, disrupts skin barrier function in neonates of varying gestational ages. PMID- 9329412 TI - Outcome for children treated with fetal intravascular transfusions because of severe blood group antagonism. AB - OBJECTIVE: To describe the outcome for 92 fetuses treated between May 1987 and January of 1993 with intrauterine (intravascular) transfusions for severe hemolytic disease in comparison with a high-risk and a healthy control group. STUDY DESIGN: Information on the perinatal period was obtained from the patient records. The children regularly attended the outpatient clinic, and a general pediatric examination was performed on each visit. The psychometer development of the child until age 4 1/2 years was assessed according to Gesell. At the age of 5 years, the adaptation part of the Denver Developmental Screening Test and a Dutch language test were used. A neurologic examination was performed according to Touwen. RESULTS: In our study, 77 (83.7%) of 92 fetuses were born alive after intravascular transfusions. The overall survival rate was 79.3%. The follow-up group included 69 infants, with an age range of 6 months to 6 years. Correlation between antenatal and perinatal features showed a significant negative relationship between the number of intrauterine transfusions and the duration of phototherapy (p = 0.002). The probability that neurologic abnormalities would occur was significantly greater when perinatal asphyxia had been present (p < 0.05) and with a lower cord hemoglobin level at birth (p = 0.03). The total number of children with disabilities was 10.1% (7/69). CONCLUSIONS: The neurodevelopmental outcome for the group of survivors compared favorably with a group of high-risk, very low birth weight infants (10.1% to 18%), and less favorably with a healthy control group (10.1% to 6%). PMID- 9329413 TI - Corticosteroids decrease exhaled nitric oxide in children with acute asthma. AB - OBJECTIVES: Nitric oxide (NO) produced in human airways seems to have both homeostatic and proinflammatory actions in the respiratory system. NO production has been shown to be higher in the exhaled air of asthmatic adults than in normal subjects. The aim of this study was to evaluate exhaled NO production during asthma exacerbation in children and the effect of a rescue course of oral steroid therapy. STUDY DESIGN: We measured NO in the exhaled air of 16 children (8 girls and 8 boys, aged 6 to 13 years) with an acute asthmatic episode before and after 5 days of therapy with prednisone, and in 16 healthy children. To measure NO, children inhaled NO-free air and, breathing at tidal volume, exhaled in a circuit from which a chemiluminescence analyzer sampled continuously. To assess the effect of acute changes in bronchial caliber on exhaled NO levels, we measured NO before and after a positive bronchodilation test result with albuterol in seven children with asthma whose disease was stable. RESULTS: In the group with acute asthma (forced expiratory volume in 1 second 62% +/- 4.4% predicted, mean +/- SEM), NO levels were significantly higher (31.3 +/- 4.2 parts per billion [ppb]) than in healthy children (5.4 +/- 0.4 ppb, p < 0.001). Administration of prednisone (1 mg/kg per day orally) for 5 days resulted in a mean decrease of 46% +/- 4% in exhaled NO concentrations (16.5 +/- 2.3 ppb, p < 0.001) compared with baseline, accompanied by a significant improvement in lung function (forced expiratory volume in 1 second 90.7% +/- 4.3% predicted). However, in patients with asthma exhaled NO levels remained significantly higher than in control children (p < 0.001) after steroid treatment. When exhaled NO was measured before and after a positive result after bronchodilator reversibility testing, we found no difference in exhaled NO levels (24 +/- 3.8 ppb vs 23.8 +/- 3 ppb; difference not significant). This demonstrates that inhaled albuterol and acute changes in bronchial caliber do not affect exhaled NO measurement. CONCLUSIONS: These data show that children with asthma exacerbation have high levels of exhaled NO that rapidly decrease with oral steroid therapy. We suggest that measurement of exhaled NO may represent a noninvasive method of monitoring airway inflammation in children with asthma. PMID- 9329415 TI - Clinical utility of the polymerase chain reaction for diagnosis of enteroviral meningitis in infancy. AB - OBJECTIVE: To determine the utility of polymerase chain reaction (PCR) assay of cerebrospinal fluid (CSF), serum, and urine for rapid diagnosis of enteroviral meningitis in infants 3 months of age and younger. STUDY DESIGN: We identified prospectively infants 3 months of age and younger coming to the emergency department with fever whose examination included a lumbar puncture, blood culture, or both. Samples of CSF, serum, urine, throat, and stool specimens were collected for viral culture and, with the exception of stool, for PCR assay. Those infants who had not received prior antibiotic therapy and had sterile bacterial cultures of CSF, blood, and urine were selected for the present analysis. RESULTS: A total of 259 specimens for viral culture and 203 specimens for PCR assay were collected from 64 infants. Comparison of results of PCR assay of CSF with viral culture, the gold standard for diagnosis of enteroviral meningitis, demonstrated a sensitivity of 100% and a specificity of 90%. Because enteroviruses are not always detectable by culture, the following modified standard was established to define enteroviral meningitis: either CSF pleocytosis, sterile bacterial cultures and detection of an enterovirus in stool culture or positive viral culture of CSF, or both. With this modified definition, the sensitivity and specificity of the PCR assay of CSF were 92% and 94%, respectively. PCR assay of serum and urine offered no benefit over PCR assay of CSF alone for diagnosis of meningitis. CONCLUSION: PCR assay of CSF is useful for the rapid and reliable diagnosis of enteroviral meningitis. Application of this technique in the clinical setting can potentially diminish unnecessary hospitalization and use of antibiotics. PMID- 9329414 TI - Etiologic classification of severe hypospadias: implications for prognosis and management. AB - OBJECTIVE: Classification of severe hypospadias employing a broad array of diagnostic tools. Standardization of a diagnostic approach to children with hypospadias. Indentification of patients at risk of having malignancies and endocrine problems. DESIGN: Retrospective analysis of patients in a single-center study. SUBJECTS: Thirty-three patients with severe (scrotal or penoscrotal) hypospadias, aged 1 to 18 years. METHODS: Clinical assessment, ultrasonography, karyotyping, endocrine evaluation including adrenal steroid concentrations, sex hormone-binding globulin test for androgen sensitivity, human chorionic gonadotropin stimulation with determination of testosterone and dihydrotestosterone concentrations to exclude 5 alpha-reductase deficiency, and molecular genetic analysis of the androgen receptor gene and the 5 alpha reductase gene. RESULTS: In 12 patients the cause was clarified. Diagnoses included Drash syndrome with Wilms tumor in infancy (3 patients), partial androgen insensitivity resulting from androgen receptor mutations (2), true hermaphroditism (2), chromosomal aberration (1), deficiency of antimullerian hormone (1), gonadal dysgenesis (1), partial 5 alpha-reductase deficiency caused by a novel point mutation (1), and XX-male syndrome (1). Twelve patients had associated findings such as cardiac malformations (3 patients), rectal atresia (1), dilation of urinary tract (2), cystinuria (1), and others. CONCLUSIONS: Patients with severe hypospadias should be submitted to a standardized set of diagnostic procedures in infancy. A stepwise diagnostic study avoids unnecessary, invasive, and expensive testing. A high proportion of classified causes can be expected. Patients at risk of having malignancies or hormonal disorders must remain under close surveillance. PMID- 9329416 TI - Incidence of bacteremia in infants and children with fever and petechiae. AB - OBJECTIVE: We determined the incidence of serious invasive bacteremia caused by Neisseria meningitidis and other organisms in febrile infants and children with a petechial rash. Further, we studied the diagnostic value of laboratory and clinical finding in these patients. STUDY DESIGN: We conducted this prospective cohort study in the emergency department of an urban pediatric teaching hospital, during an 18-month period, and enrolled consecutive patients with temperature of 38 degrees C or higher and petechiae. Our measures included (1) laboratory tests (leukocyte count, coagulation profile, blood culture, and cerebrospinal fluid bacterial culture); (2) a questionnaire requesting clinical data including general appearance, number and location of petechiae, and presence or absence of purpura; and (3) a follow-up telephone survey documenting health status. RESULTS: A total of 411 patients were enrolled, with 57.7% between 3 and 36 months of age. Eight patients (1.9%) had bacteremia or clinical sepsis. Six had serious invasive bacteremia: N. meningitidis (two patients), group A streptococcus (one), or sepsis with negative culture results (three). Two had occult bacteremia caused by Streptococcus pneumoniae and no evidence of sepsis. No patient had a positive cerebrospinal fluid culture result. None of the 357 well-appearing patients (95% confidence interval: 0.0%, 1.0%) had serious invasive bacteremia. Fifty-three patients appeared ill, including all six with serious invasive bacteremia. Ill appearance of the child had a sensitivity of 1.00 (95% confidence interval: 0.60, 1.00), and a leukocyte count of 15,000 or greater, or of less than 5000, had a sensitivity of 1.0 (95% confidence interval: 0.53, 1.00) for detecting serious invasive bacteremia. All children with meningococcemia had purpura. CONCLUSIONS: Invasive bacteremia occurred less frequently in our study than in previous series and was identified by clinical criteria. Our data support the treatment of selected well-appearing children with fever and petechiae as outpatients. PMID- 9329418 TI - Pulmonary dysfunction and reduced exercise capacity in patients with myelomeningocele. AB - OBJECTIVE: To evaluate pulmonary function and exercise capacity in children with myelomeningocele. STUDY DESIGN: Prospective evaluation in a randomly selected cohort of 12 subjects (10 to 17 years of age) with myelomeningocele and 12 control subjects matched for age, sex, and arm span. METHODS: Spirometry, lung volumes, maximum respiratory pressures, maximum oxygen expenditure during arm ergometry, and anaerobic threshold were measured. RESULTS: Mean total lung capacity and fractional lung volumes were significantly lower in case subjects than control subjects. Eleven subjects (92%) had a reduced forced vital capacity; seven (58%) had restrictive disease as evidenced by reductions in total lung capacity with normal or increased forced expiratory volume in 1 second/forced vital capacity ratio. Nine subjects (75%) had respiratory muscle weakness as evidenced by reduced maximum respiratory pressures or a low maximum voluntary ventilation. Exercise capacity was reduced as evidenced by a lower maximum oxygen consumption at peak exercise (13.8 +/- 4.8 vs 21.3 +/- 7.5 ml/min per kilogram of body weight; p < 0.02) and a lower anaerobic threshold (12.4 +/- 5.1 vs 17.3 +/- 4.2 ml/min per kilogram; p < 0.01) than the control group. Though the majority of subjects with myelomeningocele had a significant degree of restrictive disease, respiratory muscle weakness, or both, only one subject had pulmonary symptoms during exercise. CONCLUSIONS: Though most subjects with myelomeningocele had a significant degree of restrictive lung disease, respiratory muscle weakness, or both, exercise capacity was mostly limited by arm weakness. Skeletal muscle weakness may mask the symptoms of an underlying pulmonary abnormality, which may not be evident unless a pathologic cause of increased ventilation is present. Pulmonary function testing is suggested to screen for these abnormalities. PMID- 9329419 TI - Minitympanometry in detecting middle ear fluid. AB - OBJECTIVE: To assess the time needed to perform tympanometry, the success rate and the importance of the child's cooperation for the accuracy of minitympanometry in detecting middle ear fluid, and the relation between the static admittance of the tympanogram and the weight of the middle ear fluid. STUDY DESIGN: Two series of patients were enrolled. The first consisted of 206 consecutive children (mean age 4.7 years, range 1 month to 16 years) from the Outpatient Emergency Department of Pediatrics in the University of Oulu; the second group consisted of 162 children (age range 7 months to 8 years) who were referred to the Department of Otolaryngology for adenoidectomy, tympanostomy, or both procedures. In the first series the success rate and the time needed to complete a minitympanometric examination on each ear were recorded. In the second series, the tympanograms were evaluated according to the cooperation of the children at the time of the tympanometric examination, and the weight of the middle ear fluid was measured and compared with the static admittance of the minitympanometric curve. Sensitivity and specificity values were calculated separately for cooperative and uncooperative patients. RESULTS: In the first series, the mean time needed for tympanometry was 2.1 minutes (range 0.5 to 10 minutes), and 179 (86.9%) of the patients were cooperative. In the second series, the sensitivity and specificity calculated for tympanometry in detecting middle ear fluid were 79% and 93% among the cooperative children. In the uncooperative group, sensitivity and specificity were 71% and 38%, respectively. The weight of the middle ear fluid varied from 5 mg to 695 mg. There was a significant negative correlation (r = -0.66, p < 0.001) between the static admittance in minitympanometry and the weight of the middle ear fluid. CONCLUSION: Minitympanometry can be done quickly, it fails rarely, and in cooperative patients it is a better tool than has been earlier suggested, but it is useless in uncooperative children. The amount of middle ear fluid varies notably even among young children. PMID- 9329420 TI - Low levels of physical activity in 5-year-old children. AB - As the prevalence of obesity in Western societies has increased to disturbing levels, interest in the role of physical inactivity in promoting this trend has increased. We assessed physical activity energy expenditure (AEE) in 127 5-year old children, 43 of whom were white children and 84 Pima Indian children; the latter group represents a population with an extremely high prevalence of obesity. Total energy expenditure (TEE) and resting metabolic rate (RMR) were measured by the doubly labeled water method and indirect calorimetry, respectively. From these measured values, different indexes of physical activity were calculated, including AEE = TEE-(RMR + 0.1 x TEE) and physical activity level (PAL = TEE/RMR). By the age of 5 years, Pima Indian children were significantly heavier (23.0 +/- 5.3 kg vs 19.1 +/- 2.6 kg) and fatter (30 +/- 7% vs 21 +/- 5% body fat) than white children (p < 0.0001), whereas TEE (5996 +/- 1005 kJ/day vs 5690 +/- 760 kJ/day) and RMR (4431 +/- 625 kJ/day vs 4236 +/- 534 kJ/day) were similar in the 2 groups in both absolute values and after adjustment for fat-free mass, fat mass, and sex. Both white and Pima Indian children had physical activity levels 20% to 30% lower (PAL = 1.35 +/- 0.13) than currently recommended by the World Health Organization (1.7 to 2.0). However, the different calculated indexes of physical activity were comparable in the two racial groups. Differences in TEE or AEE are unlikely to explain the obesity seen in Pima Indian children at a later age, suggesting that excess food intake is likely to play a major role in the cause of obesity in this obesity-prone population. However, both white and Pima Indian children have surprisingly low levels of physical activity, a condition that portends poorly for the prevention of obesity in adulthood. PMID- 9329417 TI - Successful use of a chicken-based diet for the treatment of severely malnourished children with persistent diarrhea: a prospective, randomized study. AB - OBJECTIVE: To evaluate the efficacy of a chicken-based diet for the treatment of persistent diarrhea in severely malnourished children. STUDY DESIGN: Prospective, randomized, double-blind study that compared a chicken-based diet with elemental (Vivonex) and soy (Nursoy) diets. Hospitalized children with third-degree malnutrition and persistent diarrhea, aged 3 to 36 months, were included. Diets were isocaloric and given nasogastrically at 150 ml/kg per day in progressively increasing concentrations. RESULTS: Fifty-six children were included (18 received Vivonex, 19 Nursoy, 19 chicken). They had a mean age of 6.4 +/- 4.4 months, a mean weight of 3604 +/- 1232 gm, and a mean weight-for-age percentage of 51.4% +/ 7.2%. Sixty-four percent had associated conditions on admission to the hospital. Forty-one children (73.2%) were successfully treated (13 Vivonex, 13 Nursoy, 15 chicken). There were no differences in diarrheal outcomes, and all groups had significant weight gain. Failure was independent of the diet and was associated with the presence of infection on admission. There was a significantly higher nitrogen balance in the children from the chicken group (358.2 +/- 13 mg/kg per day) than in those receiving Vivonex (226.6 +/- 61) or Nursoy (291-4 +/- 111.6; p < 0.05) groups. CONCLUSIONS: The chicken-based diet was as effective as Vivonex or Nursoy. It is well tolerated, inexpensive, and widely available and thus represents an effective and inexpensive alternative to the treatment of severely malnourished children with persistent diarrhea. PMID- 9329421 TI - Human development index as a predictor of infant and maternal mortality rates. AB - OBJECTIVE: The United Nations Human Development Index (HDI) is a composite index of life expectancy, literacy, and per capita gross domestic product that measures the socioeconomic development of a country. We estimated infant and maternal mortality rates in the world and assessed how well the HDI and its individual components predicted infant and maternal mortality rates for individual countries. MATERIALS: Data on mortality rates and values for HDI components were obtained from the United Nations and the World Bank. RESULTS: For the 1987 to 1990 period, approximately 9 million infant deaths and 349,000 maternal deaths occurred in the world annually, yielding global infant and maternal mortality rates of 67 per 1000 and 250 per 100,000 live births, respectively. HDI is a powerful predictor of both infant and maternal mortality rates. It accounts for 85% to 92% of the variation in infant mortality rates, and 82% to 85% of the variation in maternal mortality rates among countries. Each component of HDI is also strongly correlated with both infant and maternal mortality rates (significance of all values for r, p < 0.001), and eliminating life expectancy from HDI does not decrease significantly the predictive power of HDI for infant or maternal mortality rates. CONCLUSION: HDI is not only a useful measure for socioeconomic development, but also a powerful predictor of infant and maternal mortality rates for individual countries. PMID- 9329422 TI - Topical iodine-containing antiseptics and subclinical hypothyroidism in preterm infants. AB - The influence of topical iodine-containing antiseptics on thyroid function test results of premature infants was determined in two separate studies. Thyroxine and thyrotropin levels were measured on blood-spotted filter paper. Samples were obtained from 128 premature infants on their tenth day of life; the infants were treated in two neonatal intensive care units. Both units used similar treatment protocols; however, one routinely used topical iodinated antiseptic agents (n = 73), whereas the other used chlorhexidine-containing antiseptics (n = 55). There was no difference in the mean T4 levels between the two groups. The mean thyrotropin levels were elevated in preterm babies exposed to iodine (15.4 vs 7.8 mIU/L, p < 0.01). Among the iodine-exposed infants, elevated thyrotropin levels (> 30 mIU/L) were found in 13.7% of infants, compared with none in the chlorhexidine-treated group (p < 0.01). We then studied an additional 46 premature infants who were treated in one neonatal intensive care unit. Iodine containing solutions were used in 24 infants and chlorhexidine was used in 22 infants. T4 and thyrotropin levels were measured weekly during the first 28 days, one every 2 weeks until the age of 60 days, and at the age of 90 days. Among iodine-exposed infants, 20.8% had thyrotropin values > 30 mIU/L, whereas none of the infants in the chlorhexidine group had elevated thyrotropin values (p < 0.05). The elevated thyrotropin levels correlated positively with the area of disinfection. Elevated urine iodine levels were present reflecting an abnormally high iodine absorption. This study suggests that iodine absorption from topical iodine-containing antiseptics may cause disturbances in thyroid function test results in premature infants. We recommend that caution be exercised in the use of iodine-containing antiseptics in premature infants. PMID- 9329423 TI - Neonatal onset of hyperornithinemia-hyperammonemia-homocitrullinuria syndrome with favorable outcome. AB - We report on a neonate with hyperammonemic coma in whom hyperornithinemia hyperammonemia-homocitrullinuria syndrome was diagnosed. Appropriate treatment led to rapid clinical and metabolic improvement. The incorporation of 14C ornithine on cultured fibroblasts confirmed the diagnosis. At the age of 18 months, the patient is in excellent clinical condition. PMID- 9329424 TI - Very long chain acyl-coenzyme A dehydrogenase deficiency in two siblings: evolution after prenatal diagnosis and prompt management. AB - A boy had neonatal seizure, lethargy, and metabolic acidosis at presentation. He recovered completely, but the recurrence of a similar episode with associated cardiomyopathy and dicarboxylic aciduria at 10 months of age led to the recognition of a fatty acid oxidation defect. A diagnosis of very long chain acyl coenzyme A dehydrogenase deficiency was later made by enzyme assay in culture fibroblasts from this child, as well as in cultured amniotic cells from a sibling fetus. This prenatal diagnosis forestalled neonatal injury by close clinical and metabolic monitoring of the second infant. Early diagnosis and management should potentially improve the generally poor prognosis for patients with very long chain acyl-coenzyme A dehydrogenase deficiency. PMID- 9329425 TI - Segregation of the G8993 mutant mitochondrial DNA through generations and embryonic tissues in a family at risk of Leigh syndrome. AB - We identified the T8993G mitochondrial mutation in a female infant who died of Leigh syndrome. The proportion of mutant mitochondrial DNA increased to near homoplasmy in three generations of the pedigree. A similarly high proportion of mutant mitochondrial DNA was found in the chorionic villi and in fetal tissues from a pregnancy interrupted because of the risk of Leigh syndrome. This study supports the concept that prenatal diagnosis can be used for Leigh syndrome with the T8993G mitochondrial DNA mutation. PMID- 9329426 TI - Concomitant administration of sodium dichloroacetate and vitamin B1 for lactic acidemia in children with MELAS syndrome. AB - Myoclonic seizures, intractable abdominal pain, and headaches resolved during the concomitant administration of sodium dichloroacetate and vita min B1 in two Japanese siblings with the MELAS syndrome (mitochondrial myopathy, encephalopathy, lactic acidosis and strokelike syndrome). PMID- 9329427 TI - Immunoablation does not delay the neurologic progression of X-linked adrenoleukodystrophy. AB - We report the results of a near total myeloablation in preparation for bone marrow transplantation in a boy with minimal symptoms of X-linked adrenoleukodystrophy. Severe cerebral X-linked adrenoleukodystrophy developed in the patient after failure of bone marrow transplantation. This experience suggests that immunotherapy alone is not responsible for the improvement observed in some patients with X-ALD after BMT. PMID- 9329428 TI - Concentrations of biotin metabolites in human milk. AB - Because estimates of the biotin requirement for infants currently are based on the biotin concentration in human milk, we sought to determine whether inactive biotin metabolites are present. Samples were collected for 7 weeks post partum from 15 healthy women. Biotin and the inactive metabolites bisnorbiotin and biotin sulfoxide were measured by means of a high-performance liquid chromatography avidin-binding assay. At 8 days post partum the proportion of biotin was 44%, bisnorbiotin 48%, and biotin sulfoxide 8%. Although biotin content increased post partum (p < 0.003), the metabolites remained an important portion of the total providing evidence that accurate measurement of biotin in human milk requires an assay that is specific for biotin. PMID- 9329430 TI - Clinical significance of cytokine measurement for detection of meningitis. AB - Levels of interleukin-6 and tumor necrosis factor alpha were measured in cerebrospinal fluids from patients with meningitis. Interleukin-6 was increased in aseptic and bacterial meningitis, whereas tumor necrosis factor alpha was increased only in bacterial meningitis. We concluded that measurement of cytokines in cerebrospinal fluid may be useful for the rapid diagnosis of meningitis. PMID- 9329429 TI - Necrotizing fasciitis after Plastibell circumcision. AB - Necrotizing fasciitis is a potentially life-threatening infection of subcutaneous tissues and Scarpa's fascia that rarely affects neonates. We report the occurrence of this devastating infection in two neonates after routine Plastibell circumcision. These case reports highlight the presentation and management of this complication after a relatively routine and frequently performed operation. This report also emphasizes the differences between cellulitis and necrotizing fasciitis and suggests strategies for management. PMID- 9329431 TI - Staphylococcal necrotizing fasciitis in the mammary region in childhood: a report of five cases. AB - OBJECTIVE: Necrotizing fasciitis is a highly lethal soft tissue infection rarely reported in childhood. The initiating site is usually a local trauma or a surgical wound. We observed five cases of necrotizing fasciitis the initiating site for which was the mammary region and discuss their management. STUDY DESIGN: We describe these five patients and review the clinical characteristics of their presentation. RESULTS: Staphylococcal necrotizing fasciitis was observed in the mammary region in all five cases. Four children were newborn infants with a mammitis preceding the onset of necrotizing fasciitis. Surgical debridement was done only after the fourth day from onset of illness. All children were discharged in good condition after 1 month. Two have been followed until puberty, with destruction of the mammary gland in one case and good development in the other one. CONCLUSION: Mammitis may be the initiating event for necrotizing fasciitis in neonates. Necrotizing fasciitis is a life-threatening disease; patients require early intensive care, parenteral antibiotic therapy, and surgical debridement. In a few instances surgery can be carefully delayed until the necrotic area is more delineated if the condition is diagnosed early during disease evolution and appropriate treatment is instituted in intensive care units. PMID- 9329432 TI - Treatment of chronic recurrent multifocal osteomyelitis with interferon gamma. AB - Chronic recurrent multifocal osteomyelitis is characterized by recurrent episodes of painful swollen lesions of the bone and overlying skin with radiographic changes and an elevated sedimentation rate. It resembles infectious osteomyelitis but with negative findings on bacterial culture and no response to antibiotics. We treated a 13-year-old girl with interferon gamma for 3 months. She had 11 episodes of chronic recurrent multifocal osteomyelitis in 2 1/2 years before therapy and has had none in the 15 months since therapy, an outcome suggesting a favorable therapeutic response. PMID- 9329433 TI - Chronic hepatitis in an infant, in association with human herpesvirus-6 infection. AB - A 20-month-old boy was investigated for persistent liver dysfunction. Liver histologic findings showed chronic hepatitis. The presence of human herpesvirus-6 DNA in liver tissue was demonstrated both by in situ hybridization and by polymerase chain reaction. Human herpesvirus-6 may be a causative agent of chronic hepatitis in this case. PMID- 9329434 TI - Day-care centers and diarrhea: a public health perspective. AB - OBJECTIVE: To assess the relation between morbidity from acute diarrhea and the form of day care. STUDY DESIGN: The design was a retrospective cohort study. The setting was the city of Espoo, an urban-suburban municipality in southern Finland with a population of 170,000. The study population comprised 2568 randomly selected children aged 1 to 7 years. The main outcome measure was the occurrence of diarrhea. RESULTS: Children in day-care centers (DCCs) had an increased risk for acute diarrhea compared with children in home care. In the whole group of children in DCCs, the relative risk was 1.20 (95% confidence interval [CI], 1.08 to 1.34). The risk was greatest in 1- and 2-year-old children, for whom the estimated relative risks were 1.76 (95% CI, 1.28 to 2.43) and 1.56 (95% CI, 1.16 to 2.09), respectively. The proportion of diarrhea episodes attributable to DCC care in 1-year-old children was 49% (95% CI, 18% to 91%), in 2-year-old children 37% (95% CI, 11% to 73%), and in the whole group 17% (95% CI, 7% to 29%). The infection risk did not differ between children in home and family care. CONCLUSIONS: The results provide quantitative evidence that the care in DCCs is a major determinant of acute diarrhea in children, whereas family day care does not increase the infection risk. PMID- 9329436 TI - Recurrent immune cytopenias in two patients with DiGeorge/velocardiofacial syndrome. AB - We describe two patients with clinical and cytogenetic findings consistent with DiGeorge/velocardiofacial syndrome who had recurrent cytopenias at presentation. Our observations suggest that recurrent cytopenias may be part of the clinical spectrum of deletion 22q11.2. We also suggest that the diagnosis of DG/VCF syndrome be considered in patients with unexplained recurrent immune cytopenias in association with cardiac lesions, subtle craniofacial dysmorphisms, and/or learning or behavioral impairments. PMID- 9329435 TI - Acquired inhibitors to factors V and X after exposure to topical thrombin: interference with monitoring of low molecular weight heparin and warfarin. AB - Repeated surgical exposure to topical bovine thrombin is known to be associated with the development of antibodies to bovine and human thrombin and factor V. This is demonstrated by abnormalities of in vitro coagulation assays and, rarely, postoperative bleeding. We describe a 4-year-old child in whom an antibody to bovine factor X developed after cardiac surgery; this antibody interfered with the heparin anti-Xa assay, thereby complicating the monitoring of heparin therapy. PMID- 9329437 TI - Lymphoid hyperplasia causing recurrent rectal prolapse. AB - An 8-year-old girl had a 5-month history of recurrent rectal prolapse. On colonoscopy, two submucosal masses were noted in the distal rectum and diagnosed by biopsy as benign lymphoid hyperplasia. These were excised by limited dissection superficial to the submucosa, and the histologic diagnosis was confirmed. The child has done well after removal of the nodules, with no subsequent prolapse for more than 2 years. PMID- 9329438 TI - Serum free thyroxine concentration is not reduced in premature infants with respiratory distress syndrome. AB - OBJECTIVE: We used improved methods of assay to determine whether pituitary thyroid function is altered in premature infants with respiratory distress syndrome (RDS) during the first week of postnatal life. METHODS: Serum free thyroxine (T4) was measured by direct equilibrium dialysis, total thyroxine (TT4) by radioimmunoassay, and thyrotropin by a sensitive immunometric assay in 90 premature infants (45 healthy control subjects and 45 with RDS) during their first week of life after 25 to 30 weeks of gestation. Infants in the RDS group received exogenous surfactant therapy. RESULTS: Free T4 and thyrotropin concentrations of infants were not significantly different between RDS and control groups. As expected, infants with RDS had significantly lower serum total T4 concentrations compared with control infants (p < 0.001). This difference was present even after stratification for gestational age (25- to 27-week group, p = 0.012; 28- to 30-week group, p = 0.002). Lower total T4 concentrations were attributable to lower T4 binding to serum proteins among infants with RDS compared with control subjects, especially in the 25- to 27-week gestation group (p = 0.0075). CONCLUSION: These data indicate that pituitary-thyroid function is not altered in premature infants with RDS. The low total T4 state in these premature infants is attributable solely to reduced serum T4 binding, as is often seen in acute nonthyroidal illnesses. PMID- 9329439 TI - A teenager with pacemaker twiddler syndrome. PMID- 9329440 TI - Reimmunization to pneumococcus in patients with sickle cell disease: do they or don't they respond? PMID- 9329441 TI - Framingham Safety Survey. PMID- 9329442 TI - Bioelectrical impedance analysis for estimation of body water spaces. PMID- 9329443 TI - Treating the common cold. PMID- 9329444 TI - Repairing the shattered self: recovering from trauma. PMID- 9329445 TI - Trauma: prevalence, impairment, service use, and cost. AB - A review of the literature on the epidemiology of trauma reveals that traumatic events are common: most Americans experience at least one over the course of their lives. According to recent estimates, 5% of men and 10% to 12% of women will suffer from posttraumatic stress disorder (PTSD) sometime in their lives, and for victims of traumas such as rape, the rate may be as high as 60% to 80%. For at least a third of sufferers, PTSD is a persistent condition lasting many years. Over 80% of persons with PTSD suffer from other psychiatric disorders. Many also experience marital, occupational, financial, and health problems. While trauma victims are disproportionate users of the health care system, they are reluctant to seek mental health treatment. Consequences of exposure to trauma are enormously costly, not only to the victims, but also to our health care system and to society as a whole. PMID- 9329446 TI - Posttraumatic stress disorder and comorbidity: recognizing the many faces of PTSD. AB - Posttraumatic stress disorder (PTSD) commonly occurs with other psychiatric disorders. Data from a recent epidemiologic survey indicate that approximately 80% of individuals with PTSD meet criteria for at least one other psychiatric diagnosis. PTSD is particularly likely to be comorbid with affective disorders, other anxiety disorders, somatization, substance abuse, and dissociative disorders. Comorbidity may affect the presentation and clinical course of PTSD. Because of the relative frequency of traumatic events and the heterogeneity of presentation of PTSD, screening for traumatic events and PTSD should be standard in both psychiatric and primary care practice. Additionally, individuals with PTSD should be screened for psychiatric comorbidity. Accurate assessment of comorbidity may be important in determining optimal psychotherapeutic and pharmacotherapeutic treatment options for individuals with PTSD. PMID- 9329447 TI - The psychobiology of posttraumatic stress disorder. AB - This review summarizes the current state of our knowledge of the psychobiology of posttraumatic stress disorder (PTSD). People with PTSD develop an enduring vigilance for and sensitivity to environmental threat. They have difficulty in properly evaluating sensory stimuli and responding with appropriate levels of physiologic and neurohormonal arousal. The inappropriate mobilization of biological emergency responses to innocuous stimuli is mirrored psychologically in an inability to properly integrate memories of the trauma and in a fixation on the past. The biological dysregulation of PTSD can be measured on physiologic, neurohormonal, immunologic, and functional neuroanatomical levels. The developmental level at which the trauma occurs affects the nature and extent of psychobiological disruptions. The availability of neuroimaging for documenting structural and functional abnormalities in PTSD has opened up new ways for understanding the neuronal filters concerned with the interpretation of sensory information in PTSD. These studies have produced a number of unexpected findings, which may alter how we conceptualize PTSD and which may force us to reevaluate appropriate therapeutic interventions. PMID- 9329448 TI - Trauma and women: course, predictors, and treatment. AB - Posttraumatic stress disorder (PTSD) resulting from aggravated assault, rape, or noncrime trauma affects over 4 million women in the United States, according to retrospective studies. Prospective studies reviewed here found that 3 months post assault the prevalence of PTSD was 48% in rape victims and 25% in nonsexual crime victims. Prolonged exposure treatment and stress inoculation training are both effective psychotherapeutic treatments for PTSD. Prolonged exposure involves having the patient relive the traumatic memory and recount the event in detail. This description is audiotaped and the patient is asked to listen to it as part of assigned homework. In vivo exposure to feared objects or situations is also assigned as homework. Stress inoculation training consists of teaching patients a variety of techniques for managing anxiety, including controlled breathing, deep muscle relaxation, thought-stopping, cognitive restructuring, preparation for stressors, covert modeling, and role-play. Both treatments have been proven to be effective alone and in combination in ameliorating chronic PTSD in women after traumatic sexual or nonsexual assault. This efficacy was maintained for 3 months of follow-up. PMID- 9329450 TI - Posttraumatic stress disorder. AB - This article reviews concepts that help synthesize the data on posttraumatic stress disorder (PTSD), a very complex condition in terms of its etiology, psychobiology, epidemiology, comorbidity, and treatment. At least four neurobiologic systems are involved in PTSD: the catecholamine, the hypothalamic pituitary-adrenocortical, the thyroid, and the endogenous opioid systems. Six other systems are probably or possibly implicated as well. The avoidance and hyperarousal of PTSD distort the patient's appraisal of the world. The symptoms of PTSD can be understood through models of learning and memory, which form the basis of behavioral treatments. The concepts of tonic and phasic alteration and of allostasis versus homeostasis also shed light on PTSD. In addition to PTSD, there may be other identifiable posttraumatic syndromes that might be diagnosed separately, such as "complex" PTSD. Cross-cultural issues may also affect clinical phenomenology and thereby confuse the diagnosis. Comorbid disorders may actually be clues to subtypes of PTSD. The fact that victims of PTSD are also more vulnerable to medical illnesses makes a closer relationship with primary care providers and other specialists mandatory. New approaches to prevention, treatment of chronic PTSD, psychotherapy, pharmacotherapy, and research hold promise of an improved prognosis for patients with PTSD. PMID- 9329449 TI - Biological therapies for posttraumatic stress disorder: an overview. AB - Both core and secondary symptoms of posttraumatic stress disorder (PTSD) respond to medication, a valuable part of overall PTSD treatment. Treatment options include antidepressants, anxiolytics, anticonvulsants, and mood stabilizers. A growing data base of results from double-blind, placebo-controlled clinical trials supports the use of antidepressants, especially tricyclics, monoamine oxidase inhibitors (MAOIs), and serotonin selective reuptake inhibitors (SSRIs). Although heightened anxiety is characteristic of PTSD, benzodiazepines have not yet proved useful in controlled trials and may be associated with a rebound effect on discontinuation. The small, open studies of anticonvulsant drugs indicate moderate to good improvement with these agents. Tricyclic, SSRI, and MAOI antidepressants have demonstrated efficacy in larger, longer-term controlled trials. Drug/psychotherapy combinations may enhance the usefulness of psychotherapeutics in the management of PTSD. Studies with tricyclics and fluoxetine indicate that magnitude and type of trauma may determine the degree of response. PMID- 9329451 TI - The future of neuroscience--an ontogeny and teleology. PMID- 9329452 TI - Consensus recommendations for the postmortem diagnosis of Alzheimer disease from the National Institute on Aging and the Reagan Institute Working Group on diagnostic criteria for the neuropathological assessment of Alzheimer disease. PMID- 9329453 TI - Molecular genetic evidence for subtypes of oligoastrocytomas. AB - The histogenesis of oligoastrocytomas remains controversial, with some data arguing similarity of oligoastrocytomas to astrocytic tumors, and other data suggesting closer relationships with oligodendroglial neoplasms. Since the molecular genetic changes in astrocytomas differ from those of oligodendrogliomas, we characterized 120 astrocytic and oligodendroglial tumors, including 38 oligoastrocytomas, for genetic alterations that occur disproportionately between astrocytomas and oligodendrogliomas, i.e. TP53 gene mutations and allelic loss of chromosomes 1p, 17p and 19q. As previously reported, TP53 mutations were common in astrocytic gliomas, occurring in approximately half of WHO grade II and III astrocytomas, but in only 5% of WHO grades II and III oligodendrogliomas. Allelic losses of chromosomes 1p and 19q, however, were common in oligodendrogliomas (41% and 63%), but less frequent in astrocytomas (9% and 35%). Oligoastrocytomas showed TP53 mutations in 12/38 (32%) cases and allelic losses of chromosomes 1p and 19q in 52% and 70%, respectively. Most importantly, TP53 mutations and allelic losses on chromosomes 1p and 19q were inversely correlated in oligoastrocytomas (p < 0.011 and p < 0.019). These data suggest the existence of two genetic subsets of oligoastrocytomas, one genetically related to astrocytomas and the other genetically related to oligodendrogliomas. Histologically, those oligoastrocytomas with TP53 mutations were more often astrocytoma-predominant, while those with chromosome 19q loss were more often oligodendroglioma-predominant. PMID- 9329454 TI - Dendritic translocation of RC3/neurogranin mRNA in normal aging, Alzheimer disease and fronto-temporal dementia. AB - RC3/neurogranin is a postsynaptic protein kinase C (PKC)-/calmodulin-binding substrate implicated in long-term potentiation (LTP) forms of synaptic plasticity. Our previous digoxigenin in situ hybridization (DIG-ISH) studies detected RC3 mRNA in apical dendrites and cell bodies of neurons in the rat cerebral cortex and hippocampus. This observation suggested that RC3 mRNA is selectively translocated to dendrites, where it may be translated locally in response to synaptic activity. To test this hypothesis further, we isolated a full-length cDNA clone of the homologous human RC3 mRNA from a human cortex lambda GT11 library, determined its nucleotide and predicted amino acid sequences, and performed mRNA expression studies in cerebral cortex from normal human patients and from patients with Alzheimer disease (AD) and fronto-temporal dementia (FTD). The human cDNA clone detects a single approximately 1.3 kb mRNA whose nucleotide sequence is 73% similar to the rat nucleotide sequence and 96% similar to its amino acid sequence. DIG-ISH studies detect robust staining of RC3 mRNA in cell bodies of numerous neurons throughout Layers II-VI and in both apical and basal dendrites of pyramidal neurons in human neocortex (temporal/frontal). We conclude that dendritic targeting of RC3 mRNA is conserved in human brain. In AD neocortex tissue, there is little or no evidence for RC3 mRNA translocation to dendrites, while in FTD neocortex, targeting of RC3 mRNA to apical dendrites is preserved. Comparative studies in AD and FTD point to the potential importance of synapse integrity and the dendritic cytoskeleton in RC3 mRNA targeting in the human neocortex. PMID- 9329455 TI - Distribution of parvalbumin-immunoreactive neurons in brain correlates with hippocampal and temporal cortical pathology in Creutzfeldt-Jakob disease. AB - There is a distinctive pattern of hippocampal involvement in Creutzfeldt-Jakob disease (CJD) and evidence for selective vulnerability of GABAergic neurons in experimental and human prion disease. We studied hippocampus and temporal cortex from human CJD and control autopsy brains and surgical cryptogenic temporal lobe epilepsy specimens for distribution and density of parvalbumin (PV) and calbindin D28K (Cal) -positive neurons that are subpopulations of GABAergic neurons. Pathology was evaluated semiquantitatively in 8 regions in 23 CJD brains for severity of spongiform change, astrogliosis and pathological prion protein deposition. In CJD, pathology was severe in pre-parasubiculum and temporal cortex, and little or absent in CA1-4; PV+ neurons were severely reduced or absent in all cases, whereas Cal+ neurons were largely preserved. In controls, the density of PV+ neurons was highest in pre-parasubiculum and temporal cortex, and lowest in CA1-4. In cTLE, loss of PV+ neurons was seen only in CA1-4. The diffuse and severe loss of PV+ neurons in CJD, and the topographical correlation of tissue lesioning in CJD with density of PV+ neurons in controls suggest selective vulnerability and early loss of this subset of inhibitory neurons in CJD. This might relate to characteristic CJD symptoms such as myoclonus and the distinctive EEG pattern. PMID- 9329456 TI - Detection of chromosomal changes by interphase cytogenetics in biopsies of recurrent astrocytomas and oligodendrogliomas. AB - Recently, lineage-specific genetic pathways of tumor progression have been suggested in both oligodendrogliomas and astrocytomas. Aberrations consistently reported in gliomas include chromosomes 1, 7, 10, 17 and 19. Identification of specific genetic damage may have important clinical consequences, because oligodendrogliomas, unlike astrocytomas, are responsive to chemotherapy. Genetic alterations specific for tumor type and tumor progression were investigated in 5 pairs of recurrent astrocytomas and 8 pairs of recurrent oligodendrogliomas by means of interphase in situ hybridization (ISH) to paraffin-embedded, formalin fixed tissue sections. A set of DNA probes specific for the centromeric regions of chromosomes 1, 7, 10, 17, X and Y was applied. Since LOH studies on oligodendrogliomas have revealed losses in the region of 1p32-1p36, a DNA probe specific for the 1p36.3 locus was included. Hybridization with the 1p36.3 probe revealed loss of 1p in 5 of the 8 oligodendroglioma recurrences, the aberration being present in the primary tumors in 2 cases. In none of the astrocytomas was loss of 1p observed. Numerical aberrations were found in one astrocytoma pair (+7) and in the second biopsy of an oligodendroglioma (+7, -10). Aneuploidy was found by in situ hybridization in 8 of the 13 tumor pairs. Detection of aberrations in the 1p36.3 locus by interphase in situ hybridization to paraffin embedded, formalin-fixed tumors may become a very useful tool in delineation of oligodendroglial from astrocytic genotypes, directing tumor specific therapy. The technique may be of crucial importance in tumor cases in which histologic criteria of lineage are not obvious. PMID- 9329457 TI - Temporal and regional patterns of axonal damage following traumatic brain injury: a beta-amyloid precursor protein immunocytochemical study in rats. AB - Diffuse axonal injury (DAI) is an important consequence of human head trauma. This experimental investigation utilized the immunocytochemical visualization of beta-amyloid precursor protein (beta-APP) to document regional patterns of axonal injury after traumatic brain injury (TBI) and to determine the importance of injury severity on the magnitude of axonal damage. Rats underwent moderate (1.84 2.11 atm) or severe (2.38-2.52 atm) parasagittal fluid-percussion (F-P) brain injury or sham procedures. At 1, 3, 7 or 30 days after TBI, rats were perfusion fixed and sections immunostained for the visualization of beta-APP. A regionally specific axonal response to TBI was documented after moderate F-P injury. Within the dorsolateral striatum, an early increase in beta-APP-positive axonal profiles at 24 hours (h) was followed by a significant decline at subsequent survival periods. In contrast, the frequency of reactive profiles was initially low within the thalamus, but increased significantly by day 7. Within the external capsule at the injury epicenter, numbers of immunoreactive axons increased significantly at 24 h and remained elevated throughout the subsequent survival periods. At multiple periods after TBI, selective cortical and thalamic neurons displayed increased staining of the perikarya. A significant increase in the overall frequency of beta-APP profiles was documented in the severe vs moderately injured rats at 72 h after TBI. These data indicate that parasagittal F-P brain injury (a) results in widespread axonal damage, (b) that axonal damage includes both reversible and delayed patterns, and (c) that injury severity is an important factor in determining the severity of the axonal response to TBI. PMID- 9329458 TI - Expression of telomerase RNA component correlates with the MIB-1 proliferation index in ependymomas. AB - Although there is general agreement that certain morphologic subtypes of ependymoma are benign, the biologic behavior of other ependymal neoplasms is poorly understood and not clearly related to conventional histopathologic criteria. The absence of universally accepted standards has prompted the search for more objective biologic markers. Telomerase is an RNA-containing enzyme associated with immortality in proliferating stem cells and many tumors. We investigated the proliferative activity of 26 ependymomas as determined by MIB-1 immunolabeling and compared the results with the in situ expression of human telomerase RNA (hTR) and WHO tumor grade. The study included 9 WHO grade I ependymomas (6 subependymomas and 3 myxopapillary ependymomas), 13 WHO grade II ependymomas, and 4 anaplastic (WHO grade III) ependymomas. The proliferation index (PI) and telomerase RNA expression were significantly increased in grade III ependymomas (p < 0.0001 for PI and p = 0.0015 for hTR). In these tumors, the PI and hTR expression were highly correlated (p = 0.0001). Of note, a single case designated grade II showed both increased proliferative activity and the highest hTR expression detected in this series of ependymal neoplasms. Our results suggest that the PI and hTR expression may be important biologic markers, independent of other histopathologic criteria of tumor grade. Future studies examining the correlation of MIB-1 cell kinetics and hTR expression with clinical parameters in selected ependymoma subtypes are needed to determine the prognostic relevance of these markers. PMID- 9329459 TI - Neuropathological findings in eight children with cerebro-oculo-facio-skeletal (COFS) syndrome. AB - Cerebro-oculo-facial-skeletal (COFS) syndrome is a rare autosomal recessive disorder with microcephaly, severe mental retardation, and death in childhood. The pathogenesis is unknown. Neuropathological features of 8 children with COFS syndrome are presented. Seven of the children, ranging in age from 36 weeks gestation to 5 years 8 months, are of North American aboriginal background from Manitoba, Canada. The eight child is a 3-year-old Caucasian male. In all children there was severe microencephaly and mild ventriculomegaly. Cerebral myelination appeared to be delayed in one infantile case. Swollen ubiquitinated granular cells appeared in the white matter shortly after birth. Older children displayed cortical neuron loss, patchy or diffuse absence of myelin and gliosis in the white matter, and pericapillary and parenchymal mineralization in the globus pallidus and to a lesser extent the putamen and cerebral cortex. The cerebellum of older children exhibited severe degenerative changes involving the internal granular layer and Purkinje cell layer. The neuropathological changes, previously not well documented, suggest that COFS syndrome is associated with a degenerative process that begins in utero and affects many brain cell types. Similarities to Cockayne syndrome are discussed. PMID- 9329460 TI - Human mucopolysaccharidosis IIID: clinical, biochemical, morphological and immunohistochemical characteristics. AB - Mucopolysaccharidosis IIID (MPS IIID) is one of the rarest of the MPS-III syndromes. To date, the clinical manifestations of 10 patients have been reported, the deficient N-acetylglucosamine 6-sulfatase (G6S) enzyme has been purified, and the G6S gene has been cloned, sequenced and localized. However, morphological manifestations of this condition have not been reported and the pathogenesis of the severe neurological deficits remains an enigma. In this paper we describe and correlate the clinical, biochemical and pathological observations for 2 cases of MPS IIID. We used monoclonal antibodies against heparan sulfate (HS) and GM2-ganglioside, thin layer chromatography, mass spectrometry, and morphological techniques to demonstrate the nature and the distribution of the uncatabolized substrates. The majority of the cells in various tissues showed morphological changes expected with lysosomal storage of HS. The central nervous system (CNS) was most severely affected because of the secondary storage of GM2 and GM3 gangliosides in addition to the primary accumulation of HS. The extent as well as the distribution of the diverse storage materials varied within and among different neurons as observed in MPS-III A, B, and C syndromes. This study supports the hypothesis that the neurological dysfunction and neurodegeneration common to the Sanfilippo syndromes is, in part, due to the secondary metabolic perturbations induced by HS accumulation. PMID- 9329461 TI - Therapy-induced alterations in host defense in children receiving therapy for cancer. PMID- 9329462 TI - Hepatoblastoma in children: a review. AB - The past 25 years have seen a dramatic improvement in results of treatment of children with HBL; formerly, < 25% were cured, and today, 65 to 75% may be cured. New, active agents are still needed, and the late effects of therapy, especially on the heart and kidneys, remain a concern. Nevertheless, it is clear that treatment of HBL is indeed "a bit of a success story" (42). PMID- 9329463 TI - Equal participation of minority patients in U.S. national pediatric cancer clinical trials. AB - PURPOSE: To determine the ethnic/racial distribution of patients entered in national pediatric cancer clinical trials relative to the patient population served. METHODS: The ethnic/racial distribution of 29,134 patients < 20 years of age entered in clinical trials conducted by the Children's Cancer Group (CCG) and Pediatric Oncology Group (POG) between January 1, 1991 and June 30, 1994 were compared with the expected distribution of patients of the same age in the United States. The latter was predicted from the 1989 to 1991 crude incidence data of the National Cancer Institute (NCI) Surveillance, Epidemiology, and End Results (SEER) program applied to the 1990 United States census. RESULTS: Of the patients on CCG and POG trials, 11.6% were reported to be Hispanic, 10.4% were African American, and 4.7% were other racial groups. The expected values were 9.1%, 10.7% and 4.3%, respectively. Representation of minority patients was equal or greater than expected for 24 of 27 subgroups analyzed. CONCLUSIONS: In the United States, minority children with cancer are proportionately represented on clinical trials of the two national pediatric cancer cooperative groups. They and their families are provided with an equal opportunity to access clinical cancer trials and the potential benefits of cancer research. PMID- 9329464 TI - Infant cancer in the U.S.: histology-specific incidence and trends, 1973 to 1992. AB - BACKGROUND: Many cancers in infants demonstrate unique epidemiologic, clinical, and genetic characteristics compared with cancers in older children. Few epidemiologic reports, however, have focused on this important age group. METHODS: Population-based data from the Surveillance, Epidemiology, and End Results (SEER) program were used to estimate relative frequency, incidence rates, and average annual percentage change of rates among children in their first year of life (infants) who were diagnosed with a malignant neoplasm from 1973 to 1992 (N = 1461). RESULTS: The greatest proportion of cases (12%) was diagnosed during the first month of life, with extracranial neuroblastoma accounting for 35% of this total. Overall, the average annual incidence rate was 223/1,000,000 infants. Extracranial neuroblastoma was the most common infant malignancy (58/1,000,000 infants per year), followed by leukemias (37/1,000,000), brain and central nervous system (CNS) tumors (34/1,000,000), and retinoblastoma (27/1,000,000). White infants had a 32% higher incidence rate than black infants. The average annual percentage increase in rates for all cancer from 1973 to 1992 was 2.9% (95% CI: 1.9%, 3.8%). For neoplasms with at least 100 cases, increasing trends were greatest for retinoblastoma (4.6%), CNS (4.1%), and extracranial neuroblastoma (3.4%). CONCLUSIONS: Incidence rates increased notably over the study period. Future studies should consider the unique presentation of infants with cancer when developing new hypotheses related to cancer etiology and gene environment interactions. PMID- 9329465 TI - Evans syndrome: results of a national survey. AB - PURPOSE: Our goal was to improve the management of Evans Syndrome, an uncommon and frequently refractory condition. We conducted a retrospective survey to assess the demography, presentation, clinical course, and treatment response of affected children. PATIENTS AND METHODS: Information was analyzed from a detailed questionnaire completed by pediatric hematologists mainly in the U.S. and Canada. We sought information regarding demographics, findings at presentation, approach to diagnosis, treatments used (with specific reference to splenectomy, corticosteroids, and intravenous immunoglobulin (IVIG)), course of the disease with emphasis on recurrences, and status at last follow-up. RESULTS: Forty-two patients (22 male, 20 female) were included in the study. The median age was 7.7 years (range 0.2-26.6 years). At presentation, thrombocytopenia (32 patients) and anemia (28) were common; neutropenia occurred in 10 and pancytopenia in 6. Patients received a median of 5 (range 0-12) modalities of treatment. Courses of IVIG and corticosteroids were given to almost all patients; responses were varied but the effects lasted as long as 2 years. Splenectomy was performed for 15 patients but the median duration of response was only 1 month. Other treatments included cyclosporine, vincristine, danazol, azathioprine, cyclophosphamide, and plasmapheresis. The course of the disease was characterized by recurrent thrombocytopenia, hemolytic anemia, and neutropenia. After a median follow-up of 3 years, 3 patients had died, 20 had active disease on treatment, 5 had persistent disease (not on treatment), and 14 had no evidence of disease. CONCLUSION: Evans syndrome is a chronic and recurrent condition which is often refractory to IVIG, corticosteroids, and splenectomy. Responses to other agents have been anecdotal and inconclusive. A prospective study involving these agents is needed to determine optimal therapeutic combinations. PMID- 9329466 TI - A phase II trial of high-dose methotrexate in previously untreated children and adolescents with high-risk unresectable or metastatic rhabdomyosarcoma. AB - PURPOSE: The outcome for children with advanced-stage rhabdomyosarcoma remains poor with contemporary treatment regimens. Evaluation of new drugs is important to improve clinical outcome. Because methotrexate has shown promising activity in the treatment of patients with recurrent rhabdomyosarcoma, we conducted a phase II trial in untreated children with advanced-stage disease to evaluate the efficacy and safety of this agent. PATIENTS AND METHODS: Fifteen patients received 1 to 4 courses of high-dose methotrexate (HDMTX, 12 g/m2). Patients then received standard multiagent chemotherapy (vincristine, dactinomycin, cyclophosphamide, ifosfamide, mesna) with cytokine support and local radiotherapy. Patients who responded to HDMTX received four additional courses of this drug during continuation therapy. RESULTS: Twelve patients were evaluable for response after 2 or more courses of HDMTX; 4 achieved a partial response (33.3%). After administration of standard chemotherapy and radiation, the estimated 2-year progression-free survival for all patients was 56% (SD 15%). The drug was well-tolerated and the most common side effects included mucositis, transient elevation of transaminases, and neutropenia. The four patients who received additional courses of HDMTX during continuation therapy had limited toxicity which included mucositis, anemia, and thrombocytopenia. CONCLUSIONS: About one-third of children with previously untreated advanced-stage rhabdomyosarcoma responded to HDMTX. Its different mechanism of action and non overlapping toxicity with other agents make HDMTX an attractive candidate for incorporation into front-line treatment regimens for rhabdomyosarcoma. PMID- 9329467 TI - Clinical features of myelokathexis and treatment with hematopoietic cytokines: a case report of two patients and review of the literature. AB - PURPOSE: To define the features and course of myelokathexis, a rare congenital neutropenia resulting from impaired release of granulocytes from bone marrow. METHODS: The clinical features, granulocyte function, lymphocyte function, and response to granulocyte colony-stimulating factor (G-CSF) of two patients (mother/son) with myelokathexis were studied. This experience and 14 previous reports lead to a composite description of myelokathexis. RESULTS: Both patients had chronic neutropenia, recurrent pulmonary infections, bone marrow consistent with myelokathexis, hypogammaglobulinemia, and elevated endogenous G-CSF. Patient 15 had normal granulocyte function, a rise in absolute neutrophil count (ANC) with epinephrine and hydrocortisone, and normal numbers of T- and B-lymphocytes; she also had numerous warts during childhood. Both patients experienced a transient increase in ANC with infection, a significant increase in ANC within 5 hours following a single dose of G-CSF, and fewer infections with daily G-CSF. CONCLUSIONS: Based on 16 cases, myelokathexis occurs more often in females and frequently affects multiple members of a family. The usual number of circulating granulocytes is low although function is normal. Mature marrow granulocytes are mobilized with infection, corticosteroids, epinephrine, G-CSF, and granulocyte macrophage colony-stimulating factor (GM-CSF). Lymphocyte number is normal but lymphocyte function is abnormal as evidenced by hypogammaglobulinemia and papillomavirus infection. PMID- 9329468 TI - Hemangiopericytoma of the liver: immunohistochemical observations, expression of angiogenic factors, and review of the literature. AB - PURPOSE: We describe an unusual case of a hemangiopericytoma in the liver of a child, review the literature, and characterize the tumor by immunohistochemistry and electron microscopy. We study the expression of basic fibroblast growth factor (bFGF) and of vascular endothelial growth factor (VEGF) in the tumor. MATERIALS AND METHODS: Clinical history and pathology were reviewed; sections of the tumor were studied by histology, electron microscopy, and immunohistochemistry using antibodies directed towards factor-XIIIa, HAM-56, bFGF and VEGF, among others. RESULTS: The expression of VEGF resembled that of "proliferating" hemangiomas; however, despite being markedly elevated in the urine, bFGF could not be unequivocally detected in the tumor. A subpopulation of factor XIIIa positive cells was identified, similar to the "interstitial" cells of the cellular hemangiomas of infancy. The nature and function of these cells remains speculative. CONCLUSIONS: Hemangiopericytomas are rare in the liver. When arising in this location in a child, they may clinically resemble a hemangioma, may express angiogenic factors in a similar fashion, and should be considered in the differential diagnosis. PMID- 9329469 TI - Internal carotid artery occlusion in a child with sickle cell disease: case report and immunohistochemical study. AB - PURPOSE: The purpose of this report is to describe the clinical and pathologic features of a patient with acute thrombosis of both internal carotid arteries leading to death. METHODS: This is a case report of special interest because of extensive brain vessel pathologic examination. RESULTS: The analysis of this case showed that the brain had suffered massive infarction and cerebral edema. The internal carotid arteries (ICAs) were occluded by acute thrombus. The arterial wall of the left ICA, studied at its distal segment, showed a small amount of intimal hyperplasia which did not cause encroachment on the lumen. Immunohistochemical stains indicated that this lesion was formed by proliferative vascular smooth muscle rather than incremental thrombus formation. CONCLUSION: Acute thrombus formation can occur in the large cerebral arteries of children with sickle cell disease in the presence of only minimal intimal hyperplasia. The intimal hyperplasia which forms the sickle related vasculopathy seen on angiography or detected by Transcranial Doppler may be more related to stimulation of smooth muscle cells than dysregulation of thromboregulation at the endothelial surface. Implications for preventive treatment are discussed. PMID- 9329470 TI - Successful treatment of life-threatening acute chest syndrome of sickle cell disease with venovenous extracorporeal membrane oxygenation. AB - PURPOSE: We describe a pediatric patient with sickle cell disease and life threatening acute chest syndrome who was successfully treated with venovenous extracorporeal membrane oxygenation (ECMO). PATIENT AND METHODS: An 8-year-old boy with sickle cell disease presented with vaso-occlusive crisis, which progressed to fulminant acute chest syndrome requiring a partial exchange transfusion and mechanical ventilation. Despite very high ventilator settings and significant barotrauma, hypoxia persisted and circulatory failure occurred. He was then successfully treated with venovenous ECMO for 11 days. One month after decannulation he had a seizure associated with abnormalities on magnetic resonance images (MRIs). His disease has been managed with a chronic transfusion program since then. Follow-up after 5 years reveals normal pulmonary function tests, a normal magnetic resonance angiogram (MRA), and above-average cognitive skills. CONCLUSION: This is the first report of a pediatric patient with acute chest syndrome successfully managed with venovenous ECMO. His course was complicated by a seizure associated with MRI abnormalities, although the outcome has been excellent. This case suggests that treatment with venovenous ECMO should be strongly considered for sickle cell patients with life-threatening acute chest syndrome, despite maximal conventional support. PMID- 9329471 TI - Successful treatment of two brothers with congenital afibrinogenemia for splenic rupture using heat- and solvent detergent-treated fibrinogen concentrates. AB - PURPOSE: This report describes life-threatening spontaneous splenic rupture in two brothers with congenital afibrinogenemia. PATIENTS: Two brothers, aged 11 and 14 years, had intra-abdominal bleeding due to splenic rupture confirmed by ECHO ultrasonography and computed tomography scans. RESULTS: Splenectomy was performed after administration of heat- and solvent-detergent treated fibrinogen concentrates. CONCLUSIONS: Hemostatic treatment for splenic rupture using heat- and solvent detergent-treated fibrinogen concentrates was effective. Careful attention must be paid to the risk of splenic rupture during the growth spurt in physically active children with this rare coagulation disorder. PMID- 9329472 TI - Cerebral metastases of alveolar rhabdomyosarcoma in an infant with multiple skin nodules. AB - PURPOSE: This report describes extremely rare cases of infantile rhabdomyosarcoma with multiple skin nodules. They are of interest not only for their anatomic sites, but also for subsequent cerebral metastases with sudden cranial hypertension. PATIENTS: Two infants had multiple skin nodules and excisional biopsy revealed alveolar type rhabdomyosarcomas. The patients were treated with tumor resection and combined chemotherapy without any clinical progression for 9 and 16 months, respectively. RESULTS: Evidence of cerebral metastases developed with sudden vomiting and convulsion as the first manifestation. In one patient, urgent radiographic examinations failed to reveal lesions except for dilated cerebral ventricles. Seven weeks after the onset of the neurologic symptoms, only Gd-DPTA-enhanced magnetic resonance imaging (MRI) revealed multiple punctate metastatic lesions hyperintense to the surrounding cerebral tissue. Despite appropriate chemotherapy, both patients had disease progression and died of central nervous system metastases. CONCLUSIONS: The authors emphasize the need to recognize the multiple cutaneous presentation of infantile rhabdomyosarcoma and the association of cerebral metastases as a potential and fatal complication. The diagnosis is facilitated by Gd-DPTA-enhanced MRI, particularly when cerebral computed tomography scans fail to disclose metastatic lesions. PMID- 9329473 TI - Pleural relapse during hematopoietic remission in childhood acute lymphoblastic leukemia. AB - PURPOSE: This report describes an isolated pleural relapse during hematopoietic remission in a child previously treated for acute lymphoblastic leukemia (ALL). PATIENT AND METHODS: An 11-year-old boy had a cough and exertional dyspnea 34 months after an initial diagnosis of ALL and 10 months after completion of therapy. Imaging studies revealed a large left pleural effusion. Bone marrow and cerebrospinal fluid studies were negative for disease at this time. RESULTS: Histopathologic examination of biopsy samples revealed cells with morphologic features of acute lymphoblastic leukemia blasts. Immunophenotyping, cytogenetic, and gene rearrangement studies confirmed the presence of a leukemic blast cell population similar to that detected at initial diagnosis. An isolated extramedullary relapse in the pleura was diagnosed. The patient underwent successful reinduction therapy and subsequently a matched unrelated donor bone marrow transplant; he died of disseminated infection in the posttransplant period. CONCLUSIONS: Unusual extramedullary sites of relapse are recognized with increasing frequency as long-term survival in childhood ALL improves. The length of the disease-free interval before relapse is felt to be of prognostic significance. Isolated relapse to the pleural space has not previously been described. The mechanism for persistence of leukemic clones in patients who appear to be in hematopoietic remission is unknown. PMID- 9329474 TI - Primitive neuroectodermal tumor of bone as a second malignant neoplasm in a child previously treated for acute lymphoblastic leukemia. AB - PURPOSE: Although rare, second malignant neoplasms (SMNs) are a devastating consequence of successful treatment of childhood cancer. The 15-year estimated risk of developing a second malignant neoplasm after treatment of childhood acute lymphoblastic leukemia (ALL) is 2.5%. Most of these neoplasms are central nervous system tumors. The risk of secondary acute myeloid leukemia has been negligible in most treatment regimens. Here, we report the first case of a primitive neuroectodermal tumor (PNET) in a patient treated for ALL. PATIENTS AND METHODS: A 15.7-year-old girl developed pain in her left leg 7 years after diagnosis of low-risk ALL. Imaging studies revealed lytic lesions in her left proximal tibia and several vertebra as well as metastatic nodules in both lungs. RESULTS: Immunocytochemical and molecular analyses led to the diagnosis of PNET. The treatment of this SMN was composed of combination chemotherapy with hematopoietic growth factor support and radiotherapy to the primary lesion and affected spine. The tumor recurred 5 months after the completion of treatment, and the patient is now undergoing salvage therapy composed of chemotherapy and radiotherapy. CONCLUSIONS: To our knowledge, this is the first report of PNET as an SMN after successful treatment of ALL. PMID- 9329475 TI - Recurrent rhabdomyosarcoma after 25 years: a possible association with estrogen and progestogen therapy. AB - PURPOSE: We describe a patient with a late recurrence of rhabdomyosarcoma and review the relevant literature. PATIENT AND METHODS: Recurrent rhabdomyosarcoma occurred in a young woman 25 years after initial presentation, with the onset of symptoms 3 months after commencing hormonal replacement therapy with estrogen and progestogen. The primary and recurrent tumors were immunocytochemically identical. The primary tumor was steroid receptor negative but the recurrent tumor was estrogen and progesterone receptor positive. DISCUSSION: Estrogen priming can stimulate synthesis of progesterone receptors which may modulate mitotic activity, which suggests a functional role for receptor positive cells in modulating cell growth when sex hormone primed, with a possible tumor induction role of sex hormone replacement therapy. CONCLUSION: The late tumor recurrence may have been induced by estrogen and progestogen treatment. PMID- 9329476 TI - Pain in the oldest-old during hospitalization and up to one year later. HELP Investigators. Hospitalized Elderly Longitudinal Project. AB - OBJECTIVE: To evaluate the pain experience of very old hospitalized patients during and up to 1 year after hospitalization. To understand the relationship of level of pain to demographic, psychological, and illness-related variables. DESIGN: Prospective cohort study. SETTING: Four teaching hospitals. PARTICIPANTS: 1266 patients at least 80 years of age in the Hospitalized Elderly Longitudinal Project (HELP). MEASUREMENTS: Pain interviews during hospitalization and 2 and 6 months later. Ordinal logistic regression was used to study the association of variables with level of pain. RESULTS: Interviews about symptoms were available for 806 (64.6% of survivors) patients during hospitalization, 614 (57.9% of survivors) at 2-months, and 416 (48.0% of survivors) at 12 months; of these, 45.8, 49.8 and 53.6%, respectively, reported pain, and 12.9% of those with pain during hospitalization were dissatisfied with its control. Multivariable analysis revealed that study hospital, admission diagnosis, depressed mood, alertness, and level of activity 2 weeks before admission were associated with pain during hospitalization, and pain reported during hospitalization, study site, patient level of activity 2 weeks before hospital admission, and patient education were associated with pain 2 months later. CONCLUSIONS: Frequency of pain among very old hospitalized patients and at follow-up is similar to that reported for other hospitalized patients. Further studies of strategies to better control pain during and after hospitalization in very old patients are needed. These studies will have to adjust for other variables associated with pain in the oldest-old. PMID- 9329477 TI - Psychiatric assessments of nursing home residents under OBRA-87: should PASARR be reformed? Pre-Admission Screening and Annual Review. AB - OBJECTIVE: As part of nursing home practice reforms, OBRA-87 mandates formal psychiatric assessments (PASARR) of nursing home residents suspected of having mental disorders, a responsibility it delegates individually to states. We describe the initial year of implementation of the PASARR process in King County, Washington, and characterize the mental disorders and mental health services needs of nursing home residents referred for psychiatric screening. DESIGN: Cross sectional study. SETTING: The 54 Medicare-certified King County nursing homes (total beds = 7013). PARTICIPANTS: All patients referred for psychiatric evaluation under PASARR (n = 510). MEASUREMENTS: A systematic, multidimensional evaluation including a semistructured psychiatric diagnostic examination, validated measures of cognitive dysfunction, depression, and global psychopathology, functional variables relevant to need for nursing home care, and selected mental health services indicators. RESULTS: Fewer than 10% of all nursing home residents were referred for psychiatric evaluation. A primary mental illness, evenly divided between psychoses and mood disorders, was found in 60% of the sample, and a psychiatric disorder associated with dementia or mental retardation was found in 25%. Six percent had complex neuropsychiatric features defying classification, and 4% had no mental disorder. Other disorders, such as substance abuse, were rare. Cognitive impairment and global psychopathology were prevalent in all diagnostic groups, and depressive symptoms were common even in patients without affective diagnoses. Eighty-eight percent of the sample were appropriately placed, based on their needs for daily care. Fifty-five percent had unmet mental health services needs. CONCLUSIONS: The PASARR referral process detected a group of seriously mentally ill, functionally disabled patients, most of whom required the level of care that nursing homes provide. Depressed and psychiatrically impaired dementia patients were underrepresented in the referral pool as measured against widely accepted prevalence figures for mental disorders in nursing home populations. The PASARR process as currently configured appears to be most efficient in identifying schizophrenic patients, who represent a small minority of nursing home residents, and the skewed sample it generates fails to provide an adequate basis for estimating overall mental health services needed in nursing homes. The PASARR process should be altered to improve referral rates for depressed and behaviorally disturbed dementia patients. PMID- 9329478 TI - Skin disorders and moisture in incontinent nursing home residents: intervention implications. AB - OBJECTIVE: To provide data needed to design an intervention trial to prevent or treat skin disorders in a high risk, incontinent nursing home population. DESIGN: The incidence and prevalence of nine common skin disorders were measured prospectively over a 60-day period using trained observers. Urinary and fecal incontinence frequency were measured over 24 hours, and mobility was measured with subjects both in and out of bed. Direct measures of skin moisture were taken with an impedance device in the presence and absence of urinary incontinence. Multiple regression analyses were used to relate the incontinence and mobility variables to the presence and development of skin disorders. SETTING: Four nursing homes. PARTICIPANTS: One hundred incontinent nursing home residents. MAIN OUTCOME MEASURES: Prospective measures of nine common skin disorders and skin moisture in four perineal regions under continent and incontinent conditions. RESULTS: All subjects had at least one skin condition identified during the 60 day data collection period. The most commonly observed skin condition was blanchable erythema, which occurred in 94% of the subjects, predominantly in the front and back regions that were closest to the urethra and rectum. Twenty-one percent of residents developed either a Stage 1 (nonblanchable erythema) or 2 pressure ulcer. All skin conditions were transient when measured every 3 weeks with the exception of blanchable erythema, which showed stability. Stage 3 or greater pressure ulcers and edema were not observed, and interrater reliability for the measure of papules was poor. Measures of urinary and fecal incontinence severity were correlated with blanchable erythema severity, and blanchable erythema and low bed mobility were predictive of pressure ulcer severity. Blanchable erythema severity was also predictive of Stage 1 and 2 pressure ulcers. Skin moisture levels in the back perineal farthest from the rectum (peripheral) were affected most by urinary incontinence. CONCLUSION: A trial to detect a 50% preventive effect on Stage 1 and 2 pressure ulcers would require that 167 subjects be monitored for 60 days. The transient nature of the skin effects require that skin be monitored at least once a week. Because blanchable erythema is so prevalent and appears to be associated with more severe skin conditions, it would make an excellent marker for beginning to assess the potential preventive effects of various interventions on the incidence of pressure ulcers and other related skin disorders in incontinent patients. It is likely that the back area peripheral to the urethra and rectum would experience the greatest benefit from an intervention trial to reduce moisture caused by incontinence. PMID- 9329479 TI - Longitudinal weight changes, length of survival, and energy requirements of long term care residents with dementia. AB - OBJECTIVE: We hypothesized that institutionalized patients with dementia, who frequently have feeding problems and require supervised and assisted feeding, would lose more weight during their residency than nondemented, independently functioning residents and have compromised survival. To test this hypothesis, we examined the survival and longitudinal changes in weight of two cohorts of institutionalized residents with dementia and compared these cohorts with a cohort of nondemented residents. We also measured the resting energy expenditures of a subset of the subjects with dementia as an indicator of their energy needs. DESIGN: A longitudinal cohort study with retrospective baseline chart review and subsequent follow-up of monthly weights and mortality over 4 years. SETTING: A 725-bed long-term care institution with specified levels of care. SUBJECTS: Two cohorts of residents with dementia, one consisting of subjects who required total care throughout their institutional stay (n = 31) and another group who did not initially require total care (n = 48); these were compared with a cohort with normal mentation who were functionally independent in their daily activities (n = 26). The total number of subjects was 105. MEASUREMENTS: Demographics, medical problems, and medications by chart review; functional and mental status evaluations; longitudinal monthly weights and mortality for the 48-month study period; and resting energy expenditures by indirect calorimetry. MAIN RESULTS: Residents with dementia had lower weights on admission and throughout their stay than nondemented, independently functioning residents, and they were more likely to have a weight loss of 10 lbs or more at some point during the 4-year study period. However, their mean weights did not change during the study period. The mean survival from admission of those demented residents who died was more than 3 years. Resting energy expenditures of women residents with advanced dementia were 12% lower than predicted from the Harris Benedict equations. CONCLUSION: Dementia is not necessarily associated with unremitting weight loss during institutionalization despite the frequent occurrence of feeding difficulties and temporary weight loss. This may be caused partly by the lower than expected resting energy expenditures and, hence, energy needs of affected residents as their dementia progresses. Demented residents weighed significantly less than nondemented, independently functioning residents throughout their institutional stay. Nevertheless, nursing staff are able to maintain weight and survival for extended periods even in very impaired residents. PMID- 9329480 TI - Determinants of functional status in healthy Italian nonagenarians and centenarians: a comprehensive functional assessment by the instruments of geriatric practice. AB - OBJECTIVE: To evaluate the physical ability and psychocognitive status of a population more than 90 years of age with regard to sociodemographic, behavioral, and biomedical variables known to affect functional status in old age. DESIGN: A survey design was used. SETTING: Emilia Romagna, Northern Italy. PARTICIPANTS: Eighty-four healthy community-dwelling subjects aged 90 to 106 years. MEASUREMENTS: Sociodemographic variables, health behavior, anthropometric indices, and serum DHEAS levels were recorded. Functional assessment was performed by instruments currently used in geriatric practice: the Mini-Mental State Examination (MMSE), Geriatric Depression Scale (GDS), and Activities of Daily Living (ADL) scale. A stepwise multiple regression analysis was performed. RESULTS: GDS scores correlated directly with MMSE scores and inversely with ADL severity scores. Poor education, institutionalization, sensory impairment, muscular mass loss, and lower DHEAS levels were the variables with the highest correlation to functional impairment. Smoking, alcohol consumption, and marital status were relatively unimportant. An inverse association was found between DHEAS levels and dependency scores of single ADLs (continence, mobility). CONCLUSION: Impaired cognitive and physical ability with no increase in depression prevalence was found in a sample of subjects more than 90 years of age free of major age-related disease. Muscular mass and DHEAS levels seem to play a role in maintaining physical independence. In turn, physical independence, as well as social and cultural factors, strongly affect the compliance of long-lived subjects with psychocognitive tests currently used in the clinical evaluation of younger old people, suggesting that these instruments are not reliable for screening for cognitive impairment and depression in the oldest old subjects. PMID- 9329481 TI - Development and testing of a process measure of nursing home quality of care. AB - OBJECTIVE: To develop and test quality of care process measures for three medical conditions of nursing home patients: fever, shortness of breath, and chest pain. DESIGN: Flowsheets designed to capture the critical elements of care for the above conditions were developed by an expert panel. Nursing home residents charts were reviewed retrospectively using the flow sheets. The reviews were translated into clinical scenarios, and the quality of care the scenarios represented was rated by an expert panel. SETTING: All nursing homes in Hennepin County, MN, that care for Medicaid patients. PATIENTS: A random sample of 1405 Medicaid nursing home residents from 1984 and 1988. MEASURES: Measures of quality of physician assessment and intervention, quality of nurse assessment and intervention, and global quality were developed and the intra- and interrater reliability were tested. The measures' validity was assessed by their ability to predict resident death. RESULTS: Intrarater reliability was measured as the correlation of the ratings of blinded duplicates. The correlation for the global scale and the four subscales ranged from .74 to .88 (P < .001 for all). Interrater reliability was tested by examining what percentage of the quality ratings were within one unit (1-5 scale) for all three raters. All three raters were within one unit for more than 72% scenarios for all scales. The subscale of quality of physician assessment was able to predict resident death when the worst episode of care (OR = .47, 95% CI(.31-.74)) or the mean episode of care (OR .54, 95% CI(.30-.99)) was used. None of the other subscales or the global measure predicted death. CONCLUSIONS: Through the use of an expert panel, measures of nursing home quality of care were developed for shortness of breath, fever, and chest pain. These measures have reasonable reliability and significant face validity. Their validity is supported further by the ability of one of the measures to predict resident death. PMID- 9329482 TI - Active voluntary euthanasia or physician-assisted suicide? AB - OBJECTIVE: To find out why Dutch general practitioners (GPs) and nursing home physicians (NHPs), and patients (according to their physician) opt for active voluntary euthanasia rather than for physician-assisted suicide, or vice-versa. DEFINITIONS: The following definitions were used in the study: Euthanasia is the intentional termination of life, by someone other than the patient, at the patient's request; physician-assisted suicide is intentionally helping a patient to terminate his or her life at his or her request. DESIGN: Two descriptive, retrospective studies. SETTING: The Netherlands. METHOD: Data were collected by means of anonymous questionnaires sent to a random sample of 521 GPs from the province of North Holland, 521 GPs from the rest of the Netherlands, and all 713 NHPs who were members of the Dutch Association of Nursing Home Physicians. Data were collected over the period 1986-1989 (inclusive) for GPs and the period 1986 June 1990 (inclusive) for NHPs. RESULTS: Forty-eight percent of the Gps, 78% of the NHPs, and about half of the patients who opted for euthanasia did so because of the physical condition of the patient. The reason GPs, NHPs, and patients gave most often for opting for physician-assisted suicide was that they wanted 'as far as possible to let the patient bear the responsibility.' CONCLUSION: In 38% of all cases for GPs and 57% of all cases for NHPs, only active voluntary euthanasia could be performed because of the patient's condition. In the other cases, where there was a choice, most GPs performed euthanasia, while most NHPs assisted in suicide. Active voluntary euthanasia was chosen primarily for medico-technical reasons, whereas physician-assisted suicide was selected primarily for moral reasons. PMID- 9329484 TI - A survey of management practices for isolated systolic hypertension. AB - OBJECTIVE: To determine the management practices of clinicians for patients with isolated systolic hypertension, with particular attention to treatment thresholds, medication choices, and target blood pressures. DESIGN: Self administered questionnaire. SETTING: Edmonton, Alberta, a large Canadian city. PARTICIPANTS: A random sample of 348 family physicians and 125 internists. MEASUREMENTS: Demographics of the respondents, first and second choice of antihypertensives, treatment thresholds, and target blood pressures for patients with isolated systolic hypertension. RESULTS: Excluding 54 nondeliverable questionnaires, a response rate of 67% (281 surveys) was obtained. The responding clinicians reported treatment thresholds and target blood pressures consistent with the evidence from randomized clinical trials and the recommendations of the Canadian Hypertension Society and the Fifth Report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure. Thiazide diuretics were recommended as first line therapy by 74% of internists and 58% of family physicians. Angiotensin converting enzyme inhibitors were the most frequently chosen second line drug (27% of internists and 45% of family physicians). CONCLUSIONS: The reported management practices of this group of clinicians are consistent with the evidence from randomized clinical trials and the recommendations of national consensus guidelines. PMID- 9329483 TI - Driving patterns and medical conditions in older women. AB - OBJECTIVES: To describe driving patterns (e.g., driving frequency) in older women drivers and to evaluate the impact of medical conditions and comorbidity on driving patterns. DESIGN: Cross-sectional examination of the association between medical conditions and driving patterns. SETTING: Population-based cohort from the Pittsburgh Center of the Study of Osteoporotic Fractures (SOF). PARTICIPANTS: A total of 1768 women aged 71 years or older. MAIN MEASUREMENTS: Driving information was obtained through a driving questionnaire, including driving status, weekly mileage, longest trip in the past year, etc. Data for demographics, lifestyle behavior, and medical conditions were collected through the SOF study. RESULTS: Among the participants, 1103 (62.3%) were current drivers, 337 (19.1%) had stopped driving, and 329 (18.6%) had never driven in their lifetime. The proportion reporting driving cessation and decline in driving amount increased with age. The prevalence of most medical conditions was higher among former drivers than in current or never drivers. Even after controlling for age and other demographic variables, fractures, heart disease, diabetes, self reported poor vision or hearing, as well as comorbidity were found to be associated independently with decreased driving amount, including driving cessation, decline in mileage, and avoiding long trips. CONCLUSION: Both individual medical conditions and comorbidity influence driving patterns in older drivers. Because it is common for older people to have several medical conditions simultaneously, comorbidity might be a more comprehensive measure of medical impact on driving. PMID- 9329485 TI - Accuracy of patient care staff in estimating and documenting meal intake of nursing home residents. AB - OBJECTIVES: To determine the accuracy of patient care staff estimates and documentation of food intake of residents in nursing homes. DESIGN: Prospective, observed, unblinded cohort study. SETTING: Three urban nursing home facilities. SUBJECTS: Staff estimation and documentation of 27 nursing home residents' meal intake. MEASUREMENTS: Actual amount consumed by 27 nursing home residents was ascertained by weighing food and caloric fluids on resident trays before and after one lunch time meal. Staff estimates and documentation of percent of meal consumed was compared with actual intake. RESULTS: Patient care staff estimates differed from actual intake by approximately 20%, and in most instances intake was overestimated. Almost one-third of the residents at risk for nutritional problems were not identified correctly by staff. Chart documentation of meal intake frequently did not reflect either actual amount of meal consumed or the staff's estimation of what was eaten. CONCLUSION: Study findings indicate that the present system used to document nursing home residents' intake is inadequate and that a more accurate mechanism or an entirely different process for identifying residents at risk for nutritional problems should be developed and implemented. PMID- 9329486 TI - Effect of genetic risk factors and disease progression on the cerebrospinal fluid tau levels in Alzheimer's disease. AB - OBJECTIVE: This study was undertaken to gain insights into the clinical utility of measuring cerebrospinal fluid tau protein (CSF-tau) to aid in the diagnosis of Alzheimer's disease (AD). SETTING: AD patients from Tohoku University Hospital, Sendai Japan were sampled. SUBJECTS AND METHODS: CSF-tau levels were examined by sandwich enzyme-linked immunosorbent assay in a total of 62 patients carrying different alpha 1-antichymotrypsin (ACT) and presenilin-1 (PS-1) polymorphic alleles. Further, the CSF-tau levels were followed up on two occasions during the progression of the disease in 17 AD patients. RESULTS: There was no evident gradient for tau protein in CSF. Neither the ACT/A allele nor the PS-1/1 allele affected the CSF-tau levels. Although CSF-tau levels changed to a variable extent over time, the CSF-tau levels were significantly increased (P < .01) during the follow-up period. Three of the AD patients demonstrated decreasing values, whereas 14 patients showed increasing values. Finally, these temporal changes in CSF-tau levels were not influenced by the apolipoprotein E epsilon 4, ACT/A or PS 1/1 alleles during the progression of AD. CONCLUSION: Regardless of the mechanisms leading to the degeneration of neurons in AD, our findings provide further evidences that monitoring CSF-tau levels may provide useful information about AD irrespective of the background of genetic risks and disease progression. PMID- 9329488 TI - Management of erectile dysfunction by the geriatrician. AB - Erectile dysfunction (ED) is the most common health disorder to afflict elderly men. Although 67% of men aged 70 years have ED, and their interest in sexual intercourse remains high, less than 5% receive adequate treatment. In this report, we review recent developments in our understanding of the pathophysiology of ED, how geriatricians can perform an office-based evaluation, and rational (evidence-based) treatment of this important disorder. PMID- 9329487 TI - Attitudes toward life-sustaining treatment of older persons in Hong Kong. AB - OBJECTIVES: There have been few studies of the attitudes of older Asians toward life-sustaining therapy. This paper presents the knowledge of and attitudes toward cardiopulmonary resuscitation (CPR) and life support in a group of subjects in Hong Kong. DESIGN: Cross-sectional, descriptive study. PARTICIPANTS: Of the 543 subjects, 382 were old-age home residents and 161 were in-patients of geriatric wards. MEASUREMENTS: Sociodemographic data, functional ability (using the Barthel Index), self-perceived health scale, knowledge of life-sustaining procedures, and subjects' preferences for such treatments were studied. They were also asked to give the most important reason for wanting or declining CPR, and to indicate who they believe should be the decision-maker(s) regarding whether they should receive life-sustaining treatment. RESULTS: Approximately 80% of old-age home residents and 60% of hospitalized patients had no knowledge of life sustaining therapy. The success rate of CPR was overestimated by older subjects, and most wished to be resuscitated. However, up to 20% changed their minds and declined CPR after they knew the true outcome of the procedure. Half of the subjects wanted life support. Univariate analysis found that advanced age and not having a spouse were associated significantly with CPR preference, whereas subjects' knowledge was associated with preference for life support. Multivariate analysis revealed that advanced age, not having a spouse, and female sex were independently associated with a tendency to decline CPR. A considerable proportion of older people wished to be involved in decision-making regarding life-sustaining treatment. CONCLUSION: Knowledge of life-sustaining procedures was poor among older people in Hong Kong compared with their counterparts in western countries. Although most older subjects wanted CPR, a number of them changed their minds after they knew the poor outcome. Therefore, older patients should be given more information about life-sustaining therapy and encouraged to take part in their treatment plans. PMID- 9329489 TI - A 63-year-old man with progressive dyspnea. PMID- 9329490 TI - Treatment of congestive heart failure in older persons. AB - OBJECTIVE: To review the management of congestive heart failure (CHF), with emphasis on older adults. DATA SOURCES: A computer-assisted search of the English language literature (MEDLINE database) followed by a manual search of the bibliographies of pertinent articles. STUDY SELECTION: Studies on the management of CHF were screened for review. Studies in older people and recent studies were emphasized. DATA EXTRACTION: Pertinent data were extracted from the reviewed articles. Emphasis was on studies involving the older persons. Relevant articles were reviewed in depth. DATA SYNTHESIS: Available data about the management of CHF, with emphasis on studies involving older people, were summarized. CONCLUSIONS: Left ventricular ejection fraction (LVEF) should be measured in all older persons with CHF. Underlying causes of CHF should be treated when possible. Precipitating causes of CHF should be treated. Older persons with CHF associated with an abnormal LVEF should be treated with a low sodium diet and with diuretics plus angiotensin-converting enzyme (ACE) inhibitors. If CHF persists, digoxin should be added to the therapeutic regimen. If CHF still persists, isosorbide dinitrate plus hydralazine should be added. If CHF still persists, a beta blocker should also be used. Calcium channel blockers should not be used. Older persons with CHF associated with a normal LVEF should be treated with a low sodium diet and with diuretics plus ACE inhibitors. If CHF persists, a beta blocker or isosorbide dinitrate plus hydralazine or a calcium channel blocker should be added to the therapeutic regimen. If sinus rhythm is present, digoxin should not be used. The role of angiotensin II receptor antagonists such as losartan in the treatment of CHF is under investigation. PMID- 9329491 TI - When will the biology of aging become useful? Future landmarks in biomedical gerontology. AB - Where should we look to find the causes of and cure for aging? This essay considers a number of scientific discoveries, not yet made, that might dramatically increase the proportion of biogerontologists doing useful work. I will consider, in turn, problems and prospects in the areas of comparative biology, mammalian and invertebrate genetics, biomarker research, caloric restriction, and clonal senescence and then conclude with a discussion of potential links between aging and late life disease. PMID- 9329492 TI - Evidence-based medicine holds the key to the future for geriatric medicine. PMID- 9329493 TI - Management of cancer pain in older patients. AGS Clinical Practice Committee. PMID- 9329494 TI - Pressure sores and urinary incontinence. PMID- 9329495 TI - The PACE program. PMID- 9329496 TI - The PACE program. PMID- 9329497 TI - Dementia with a seasonal onset secondary to carbon monoxide poisoning. PMID- 9329498 TI - Delayed diagnosis: a 78-year-old man with AIDS. PMID- 9329499 TI - A persistent fire hazard for older adults: cooking-related clothing ignition. PMID- 9329500 TI - Egg safety. PMID- 9329501 TI - "Tacrine for treatment of sleep disturbance in dementia". PMID- 9329503 TI - Psychogenic fever or psychogenic hyperthermia? PMID- 9329502 TI - Bilateral psoas abscesses in an older diabetic male [toed]. PMID- 9329504 TI - Abstract of section scientific presentations. American Academy of Pediatrics 1997 annual meeting. October 31-November 4, 1997, New Orleans, Louisiana. PMID- 9329505 TI - Knowing your genes. PMID- 9329506 TI - Combination therapies for HIV infection and genomic drug resistance. PMID- 9329507 TI - Phyto-oestrogens and breast cancer. PMID- 9329508 TI - Spread of antibiotic-resistant bacteria from acne patients to personal contacts- a problem beyond the skin? PMID- 9329509 TI - Hazards of powdered surgical gloves. PMID- 9329510 TI - The colleges, Calman, and the new deal. PMID- 9329512 TI - Risk of ocular hypertension or open-angle glaucoma in elderly patients on oral glucocorticoids. AB - BACKGROUND: Ocular hypertension and open-angle glaucoma are well-known side effects of treatment with topical ophthalmic glucocorticoids. There is uncertainty about the risk of these disorders with oral glucocorticoid therapy. METHODS: Data from the Quebec universal health insurance programme for the elderly were used to identify 9793 patients with a new diagnosis of ocular hypertension or open-angle glaucoma, or on newly prescribed treatment for these disorders (cases). 38,325 controls were randomly selected from ophthalmology patients seen in the same month and year as the case (index date). Current use of oral glucocorticoids was defined as that within 14 days of the index date. All glucocorticoid doses were converted to the equivalent amount of hydrocortisone. The case-control analysis was done by conditional logistic regression and adjusted for age, sex, systemic hypertension, diabetes mellitus, ophthalmic glucocorticoids, glucocorticoid injections, and variables related to general health. FINDINGS: The mean ages of cases and controls were similar (74.9 [SD 6.3] vs 74.7 [6.4]). The adjusted odds ratio of ocular hypertension or open-angle glaucoma for current users of oral glucocorticoids compared with non-users was 1.41 (95% CI 1.22-1.63). There was a dose-related increase in the adjusted odds ratios for current users: 1.26 (1.01-1.56) for less than 40 mg per day of hydrocortisone, 1.37 (1.06-1.76) for patients on 40-79 mg per day, and 1.88 (1.40 2.53) for patients on 80 mg or more per day. The odds ratios also increased with the duration of treatment over the first 11 months of exposure. INTERPRETATION: The use of oral glucocorticoids increases the risk of ocular hypertension or open angle glaucoma in elderly patients. In patients in this age-group who need long term treatment with high doses of oral glucocorticoids, monitoring of intraocular pressure may be justified. PMID- 9329511 TI - Randomised trial of eradication of Helicobacter pylori before non-steroidal anti inflammatory drug therapy to prevent peptic ulcers. AB - BACKGROUND: Helicobacter pylori infection is common in patients with peptic ulcers caused by the use of non-steroidal anti-inflammatory drugs (NSAIDs). But the pathogenic role of H pylori in this disease is controversial. We studied the efficacy of eradication of H pylori in the prevention of NSAID-induced peptic ulcers. METHODS: We recruited patients with musculoskeletal pain who required NSAID treatment. None of the patients had previous exposure to NSAID therapy. Patients who had H pylori infection but no pre-existing ulcers on endoscopy were randomly allocated naproxen alone (750 mg daily) for 8 weeks or a 1-week course of triple therapy (bismuth subcitrate 120 mg, tetracycline 500 mg, metronidazole 400 mg, each given orally four times daily) before administration of naproxen (750 mg daily). Endoscopy was repeated after 8 weeks of naproxen treatment or when naproxen treatment was stopped early because of bleeding or intractable dyspepsia. All endoscopic examinations were done by one endoscopist who was unaware of treatment assignment. The primary endpoint was the cumulative rate of gastric and duodenal ulcers. FINDINGS: 202 patients underwent endoscopic screening for enrolment in the trial, and 100 eligible patients were randomly assigned treatment. 92 patients completed the trial (47 in the naproxen group, 45 in the triple-therapy group). At 8 weeks, H pylori had been eradicated from no patients in the naproxen group and 40 (89%) in the triple-therapy group (p < 0.001). 12 (26%) naproxen-group patients developed ulcers: five had ulcer pain and one developed ulcer bleeding. Only three (7%) patients on triple therapy had ulcers, and two of these patients had failure of H pylori eradication (p = 0.01). Thus, 12 (26%) patients with persistent H pylori infection but only one (3%) with successful H pylori eradication developed ulcers with naproxen (p = 0.002). INTERPRETATION: Eradication of H pylori before NSAID therapy reduces the occurrence of NSAID-induced peptic ulcers. PMID- 9329514 TI - Case-control study of phyto-oestrogens and breast cancer. AB - BACKGROUND: Phyto-oestrogens are a group of naturally occurring chemicals derived from plants; they have a structure similar to oestrogen, and form part of our diet. They also have potentially anticarcinogenic biological activity. We did a case-control study to assess the association between phyto-oestrogen intake (as measured by urinary excretion) and the risk of breast cancer. METHODS: Women with newly diagnosed early breast cancer were interviewed by means of questionnaires, and a 72 h urine collection and blood sample were taken before any treatment started. Controls were randomly selected from the electoral roll after matching for age and area of residence. 144 pairs were included for analysis. The urine samples were assayed for the isoflavonic phyto-oestrogens daidzein, genistein, and equol, and the lignans enterodiol, enterolactone, and matairesinol. FINDINGS: After adjustment for age at menarche, parity, alcohol intake, and total fat intake, high excretion of both equol and enterolactone was associated with a substantial reduction in breast-cancer risk, with significant trends through the quartiles: equol odds ratios were 1.00, 0.45 (95% CI 0.20, 1.02), 0.52 (0.23, 1.17), and 0.27 (0.10, 0.69)--trend p = 0.009--and enterolactone odds ratios were 1.00, 0.91 (0.41, 1.98), 0.65 (0.29, 1.44), 0.36 (0.15, 0.86)--trend p = 0.013. For most other phytoestrogens there was a reduction in risk, but it did not reach significance. Difficulties with the genistein assay precluded analysis of that substance. INTERPRETATION: There is a substantial reduction in breast-cancer risk among women with a high intake (as measured by excretion) of phyto-oestrogens particularly the isoflavonic phyto-oestrogen equol and the lignan enterolactone. These findings could be important in the prevention of breast cancer. PMID- 9329513 TI - HIV-1 viral load, phenotype, and resistance in a subset of drug-naive participants from the Delta trial. The National Virology Groups. Delta Virology Working Group and Coordinating Committee. AB - BACKGROUND: The Delta trial showed that combination therapy (zidovudine plus didanosine and zidovudine plus zalcitabine) substantially lengthened life and reduced disease progression compared with zidovudine monotherapy. We did a nested virological study in three countries (France, the Netherlands, and the UK) to investigate changes in markers for viral load and antiretroviral-drug resistance during therapy. METHODS: 240 zidovudine-naive HIV-1-infected patients were randomly assigned zidovudine only (n = 87), zidovudine plus didanosine (n = 80), or zidovudine plus zalcitabine (n = 73). Viral load in peripheral-blood mononuclear cells and plasma was measured by quantitative culture. Plasma HIV-1 RNA was measured by reverse-transcriptase PCR amplification, and serum p24 antigen by ELISA. Resistance to antiretroviral drugs was measured phenotypically by culture and genotypically by detection and quantification of drug-related point mutations in the pol gene. Analyses were done by intention to treat. FINDINGS: The reduction in viral load was greatest 4-12 weeks after the start of therapy and was most pronounced in the combination-therapy study groups (median reductions of RNA at 4 weeks 1.58, 1.28, and 0.49 log10 copies/mL for zidovudine plus didanosine, zidovudine plus zalcitabine, and zidovudine only, respectively). RNA levels at 8 weeks were predictive of disease progression and death after allowance for baseline values. At 48 weeks, the proportion of participants with phenotypic zidovudine resistance was similar in all three groups: didanosine and zalcitabine resistance were rare; zidovudine genomic resistance correlated with phenotypic resistance (r = 0.54, p < 0.0001) and developed earlier in the combined-therapy groups. However, participants in the zidovudine monotherapy group had higher circulating loads of resistant virus than those in the combined therapy groups. INTERPRETATION: Combined antiretroviral therapy was more efficient at lowering virus load than monotherapy. Although zidovudine resistance was common in monotherapy and combined-therapy groups, circulating concentrations of resistant virus were substantially lower in the combination groups, which is likely to be a result of the continued antiviral activity of didanosine or zalcitabine. PMID- 9329516 TI - A breathless woman with hepatosplenomegaly for 12 years. PMID- 9329517 TI - Stool elastase as a diagnostic test for pancreatic function in children with cystic fibrosis. PMID- 9329515 TI - Association between beta 2-adrenoceptor polymorphism and susceptibility to bronchodilator desensitisation in moderately severe stable asthmatics. AB - BACKGROUND: In-vitro studies have suggested that polymorphisms of the beta 2 adrenoceptor may influence the desensitisation induced by beta 2-agonists. We investigated the influence of beta 2-AR polymorphism on the development of bronchodilator desensitisation in asthma patients. METHODS: We carried out an analysis of 22 moderately severe stable asthmatics, mean age 38 years, FEV1 63% of predicted and FEF25-75 38% of predicted, who received a median inhaled corticosteroid dose of 1000 micrograms/day. Patients were randomly assigned inhaled placebo or inhaled formoterol 24 micrograms bid for 4 weeks each in a crossover study. Bronchodilator dose-response curves were made at the end of each treatment period by use of cumulative doses of formoterol (6-108 micrograms) with FEV1 and FEF25-75 measured 30 min after each dose, and up to 6 h after the last dose. We calculated the degree of bronchodilator desensitisation by comparing the dose-response (for maximum and 6 h) after placebo with that after formoterol, and expressed this degree as a percentage of placebo response. Patients were divided into groups according to genotype at codon 16: homozygous Arg 16 (n = 4), heterozygous Arg 16/Gly 16 (n = 8), and homozygous Gly 16 (n = 10). At codon 27: homozygous Gln 27 (n = 5), heterozygous Gln 27/Glu 27 (n = 11), and homozygous Glu 27 (n = 6). FINDINGS: We found a significantly (p < 0.05) greater degree of bronchodilator desensitisation with homozygous Gly 16 than with homozygous Arg 16 for maximal FEV1 response: -8% (Arg 16) vs 46% (Gly 16); and for maximal FEF25-75 response: -32% (Arg 16) vs 74% (Gly 16; 95% CI 15-92% and 49-164%, respectively). Bronchodilator responses at 6 h were also significantly (p < 0.05) different for FEV1 and FEF25-75 when Arg 16 and Gly 16 were compared and values for heterozygous Arg 16/Gly 16 were intermediate. There was significantly greater desensitisation with Glu 27 than with Gln 27 for maximal FEF25-75 response: -7% (Gln 27) vs 68% (Glu 27), p = 0.05; and for 6 h FEF25-75 response: 43% (Gln 27) vs 93% (Glu 27), p < 0.05 (95% CI 2-147% and 5-94%, respectively). All patients who were homozygous Glu 27 were also homozygous Gly 16. INTERPRETATION: We have found preliminary evidence that beta 2-adrenoceptor polymorphism is associated with altered beta 2-adrenoceptor expression in asthma patients. The homozygous Gly-16 form was significantly more prone to bronchodilator desensitisation than Arg 16, with the influence of Gly 16 dominating over any putative protective effects of Glu 27. PMID- 9329519 TI - Methaemoglobinaemia after inhalation of nitric oxide in infant with pulmonary hypertension. PMID- 9329518 TI - No Borna disease virus-specific RNA detected in blood from psychiatric patients in different regions of Germany. The Bornavirus Study Group. PMID- 9329520 TI - Mycophenolate mofetil for severe autoimmune haemolytic anemia. PMID- 9329521 TI - Transient population bypassed by polio vaccination programmes in Yunnan Province, China. PMID- 9329522 TI - Vascular headache due to intracranial meningioma: a curable form of headache. PMID- 9329523 TI - Cryptosporidiosis associated with school milk. PMID- 9329524 TI - Tick-borne lymphadenopathy--a new rickettsial disease? PMID- 9329525 TI - Regulation of the response to HIV-1 needs a combined approach. PMID- 9329526 TI - Epidemiology of childhood asthma. PMID- 9329527 TI - Centennial notions of asthma as an eosinophilic, desquamative, exudative, and steroid-sensitive disease. PMID- 9329528 TI - Generalisation from phase III clinical trials: survival, quality of life, and health economics. PMID- 9329529 TI - Chronic infections and coronary heart disease. PMID- 9329530 TI - Chronic infections and coronary heart disease. The GISSI-Prevenzione Investigators. PMID- 9329531 TI - Chronic infections and coronary heart disease. PMID- 9329532 TI - Chronic infections and coronary heart disease. PMID- 9329533 TI - Chronic infections and coronary heart disease. PMID- 9329534 TI - Screening babies for vesicoureteric reflux. PMID- 9329535 TI - Screening babies for vesicoureteric reflux. PMID- 9329536 TI - Vitamin A supplementation. PMID- 9329537 TI - Cholera and street food. PMID- 9329538 TI - Dead-in-bed syndrome in diabetes mellitus. PMID- 9329539 TI - New treatments and azathioprine in multiple sclerosis. PMID- 9329540 TI - Selective serotonin reuptake inhibitors in anorexia nervosa. PMID- 9329541 TI - Diagnosing amoebiasis. PMID- 9329542 TI - Treatment of listeria meningitis. PMID- 9329543 TI - Impact factor limits funding. PMID- 9329544 TI - Biotechnology research. PMID- 9329545 TI - Genetic counselling among Muslims: questions remain unanswered. PMID- 9329546 TI - Hypersensitivity vasculitis. PMID- 9329548 TI - First clinical results with Enzymun-Test for free PSA. AB - BACKGROUND: Prostate Specific Antigen (PSA) is widely used for the detection and monitoring of prostate cancer (CAP) but PSA is elevated in many patients with benign prostatic hyperplasia (BPH). The measurement of free PSA may improve the discrimination between CAP and BPH. MATERIAL AND METHODS: Free PSA (F-PSA) and total PSA (T-PSA) were measured using kits based on the Enzymun-Test principle. The patient population was composed of 38 untreated CAP patients, 76 BPH patients and 29 prostatitis patients. RESULTS: At cut-off levels of 0.15 for the F-PSA/T PSA ratio and 10 ng/ml for T-PSA the specificity and sensitivity for detecting CAP were respectively 87% and 84% for F-PSA/T-PSA ratio and 80% and 63% for T PSA. CONCLUSION: The F-PSA/T-PSA ratio is more powerful at discriminating between CAP and BPH than T-PSA alone and may contribute to a reduction in unnecessary invasive techniques. PMID- 9329547 TI - Cut-off value determination of CYFRA 21-1 for squamous cell carcinomas of the head and neck (SCCHN). AB - BACKGROUND: CYFRA 21-1 was found to be a sensitive and specific tumor marker especially for squamous cell carcinomas of the lung. There is no reliable tumor marker for squamous cell carcinomas of the head and neck (SCCHN) available. METHODS: The amount of Cytokeratin (Ck) 19 in healthy tissue and tissues of benign and malignant tumors from pharynx, larynx and lung was compared utilizing a dot-blot assay. CYFRA 21-1 serum levels were determined prior to therapy in 132 healthy controls, in a reference group consisting of 158 patients with benign diseases of the head and neck and in 238 patients with a histologically proven SCCHN. RESULTS: Compared to lung tissue Ck 19 levels were similar only in specimen of SCCHN. Ck 19 content in normal and benign tissue from pharynx and larynx was lower than in normal lung tissue. CYFRA 21-1 serum levels were significantly higher in patients with SCCHN compared to the control or reference group. The cut-off value for CYFRA 21-1 determined at a 95%-specificity of the reference group was found to be 2.2 ng/ml. CONCLUSIONS: The Ck 19 level in healthy tissue of the upper aerodigestive tract was lower than in normal lung tissue. Corresponding to this finding the CYFRA 21-1 serum level in the reference group with benign diseases of the head and neck was lower than that of patients with benign lung diseases. The cut-off level of CYFRA 21-1 for patients with SCCHN was determined to be 2.2 ng/ml. PMID- 9329549 TI - Tissue polypeptide-specific antigen (TPS) in pediatric malignancies. AB - Tissue Polypeptide Specific Antigen (TPS) may soon be routinely used as a proliferation marker in adult epithelial tumors. So far, no data have been available on normal or pathologic TPS values in children. Therefore the present study was designed to test the marker for the first time in pediatric malignancies. Using a commercial ELISA kit (Beki Diagnostics), serum TPS levels were determined in 270 healthy children and compared with various benign (n = 143) and malignant (n = 58) diseases. In healthy children, we found an age dependent distribution of TPS values. Median (M) TPS was found to be 105.05 U/l at birth as determined from umbilical cord blood (n = 96). By the end of the first week, the value rose to M = 164 U/l and then continuously decreased with age until reaching the adult level at around 14 years. Patients with benign masses (n = 29) did not show elevated serum TPS in contrast to children with malignancies. Advanced tumor stage, metastases, and low dignity correlated with an increase in serum values. In many cases, chemotherapy and especially surgical resection of the tumor were followed by a decrease of the previously markedly elevated TPS levels. Severe infections and impaired renal function were however related with very high values and this must be taken into account when judging the validity of TPS measurements. The marker appears useful for the differential diagnosis between benign and malignant processes. While previous investigators described its use exclusively in epithelial tumors of adults, we cannot confirm this specifity in pediatric patients: Our data show that elevated TPS values in children can also be observed in nonepithelial tumors. PMID- 9329550 TI - Prognostic relevance of p53 in node negative breast cancer. AB - In a retrospective study, immunohistochemical determinations of p53 were performed on paraffinised tissue sections of 243 patients with node negative breast cancer. These patients were primary treated in the years 1980-1986 at the UFK-Wurzburg. A complete follow up could be performed in 92% of all cases. The median follow up is at 102 month. The results have been correlated to other prognostic criteria and to the clinical course of the patients. In 77 of the 243 cases a positive immunohistochemical staining for p53 (31.7%) could be found. In 42 (17.3%) cancer specimens the p53 expression was over 5%. A significant correlation was found between p53 and tumour size (p = 0.05), ploidy (p < 0.001) and dynamic parameters of the cell cycle (S-phase-fraction, MIB1: p < 0.001). In univariate and multivariate analysis the expression of p53 did not effect disease free or over-all survival of patients with node negative breast cancer. PMID- 9329551 TI - Head and neck cancer and p53-immunogenicity. AB - BACKGROUND: p53-mutations are of major importance in the development of human malignancies and occur frequently in head and neck cancer. The detection of serum p53-antibodies has been performed for a number of different cancers. For head and neck cancer though, the occurrence of serum p53-antibodies has not been determined so far. MATERIALS AND METHODS: A set of 82 sera from patients with squamous cell carcinomas of the head and neck were screened for circulating antibodies against p53 with ELISA. RESULTS: Of 82 patients 22% (n = 18) demonstrated p53-antibodies in their sera; the specificity for malignancy was 100%. CONCLUSIONS: As far as we know, this is the first study to reveal p53 antibodies in the sera of patients with SCCHN. The high incidence of positivity for p53-antibodies in this subset of cancer patients may give additional help in the diagnosis of this often disfiguring disease. PMID- 9329552 TI - Monitoring of therapy in inoperable lung cancer patients by measurement of CYFRA 21-1, TPA- TP CEA, and NSE. AB - In a series of 381 consecutive patients with lung tumors and benign pulmonary diseases, we examined whether tumor markers CYFRA 21-1 (EIA, Boehringer, Mannheim), TPA-M (IRMA AB Sangtec Medical, Bromma, Sweden), TPS (IRMA, Beki Diagnostics AB, Bromma, Sweden), CEA and NSE (EIA, Roche, Basel) have the potential to contribute to clinical decision-making processes with respect to diagnosis and assessment of response to therapy. The sensitivity values of the marker tests in NSCLC (CYFRA 21-1: 44.4% > 3.9 ng/ml, TPA-M: 39.4% > 200 U/ml, TPS: 13.2% > 230 U/ml, CEA: 37.5% > 8.6 ng/ml), in SCLC (NSE: 61.9% > 14.0 ng/ml) and in pleural mesothelioma (CYFRA 21-1 and TPS: 36.4%) were found to be clearly inferior to the yield of standard cytopathological examinations (85-98%) when using the 95% specificity versus the group with benign pulmonary disease as cut off values. Therefore, currently available tumor markers are of minor value in the primary diagnosis of lung tumors. After curative surgery (Ro) of NSCLC only CYFRA 21-1 levels dropped to the normal range within one week. The other markers simulated residual tumor mass by displaying elevated marker levels after surgery. During the monitoring of response to chemo-/radiotherapy the changes in marker levels were compared to the clinical assessment according to standard criteria of the WHO. The criteria defined for marker response were a 65% decrease for a partial response and a 40% increase of the marker levels for progressive disease. Concordant results were obtained in 59.4% of the cases for CYFRA 21-1 (TPA-M: 63.3%, TPS: 65.5%, CEA: 54.8%, NSE: 68.9%). Most discordant results were obtained in tumor remission due to an insufficient decrease in the markers. Progressive disease was most effectively indicated by CYFRA 21-1 in NSCLC 60%) and by NSE in SCLC (70.0%). It is concluded that increasing marker levels may contribute to clinical decision making, at least in helping to decide which patients should no longer treated by ineffective and toxic drugs. PMID- 9329553 TI - Adjuvant therapy of renal cell carcinoma with active-specific-immunotherapy (ASI) using autologous tumor vaccine. AB - 116 patients with renal cell carcinoma were treated with autologous tumor vaccine after radical nephrectomy. The survival rate of these patients compared to a historical control group of 106 patients from the same hospital, which received identical surgical treatment but no adjuvant therapy was evaluated. According to defined in- and exclusion criteria there was no statistical difference between the two groups. As a consequence any significant effects resulting from the treatment with autologous tumor vaccines have to be interpreted as clinically relevant in spite of the fact that no prospective randomised design was chosen. A clear significant difference (p = 0.0007) between both groups with a benefit for the group treated with autologous tumor vaccines was observed. According to the individual Robson stages patients in Robson II (p = 0.02) and Robson III (p = 0.04) showed significantly different survival rates. Due to the short follow-up time in the group Robson I and the limited number of patients in the group Robson IV no significant difference was observed. During adjuvant treatment with autologous tumor vaccine 2 patients out of 116 showed minor side effects not exceeding WHO-grade 1. PMID- 9329554 TI - Measures to overcome HAMA interferences in immunoassays. AB - The incidence of human anti-mouse antibodies (HAMA) in different patient sera panels may be debatable, the interference of HAMA, if present, in immunochemically based assays is, however, a fact. This interference can lead to falsely elevated or depressed results depending on the nature of the HAMAs involved and the particular assay format chosen. For several reasons, assays for serum tumor markers may be especially prone to HAMA interference. Consequently, in the development of such assays, special attention should be given to the HAMA issue. In our experience, the degree of HAMA interference varies greatly from one assay to another. Nevertheless, the HAMA issue has to be addressed. Several methods have been described to remove HAMA (and other interfering substances) via sample pretreatment. Alternatively, there are also some options to counterbalance the potential effect of HAMA by using assay reagents optimized for minimal HAMA susceptibility, e.g. inclusion of an excess of non-relevant mouse antibodies. However, there is no guarantee that any given assay will not be affected by HAMA. This is especially true if a portion of the HAMA in a patient's sample is comprised of anti-idiotypic "internal image" antibodies. PMID- 9329555 TI - Basics of cancer gene therapy. AB - Gene therapy is a rapidly evolving concept for the therapy of different forms of cancer. A number of phase-I clinical trials have recently been initiated world wide. This review discusses the technical concepts underlying the protocols currently coming into clinics. Two tools essentially constitute such concepts, the vector for efficient transfer and expression of the transgene in the cancer cell, and the therapeutic gene. The most advanced vectors ready for clinical use are the retrovirus- and adenovirus-derived vectors. The application of a broad spectrum of therapeutic genes can be classified into gene replacement of replace e.g. mutated tumor suppressor genes, gene addition to increase immunogenicity by cytokine genes or introduce prodrug genes for suicide induction and, thirdly, targeted affection of gene expression by antisense oligonucleotides or expression of ribozymes e.g. directed against oncogene sequences. Cancer gene therapy holds great promise to become an important addition to the multimodality in the therapy of some forms of cancer. PMID- 9329557 TI - Rationale and clinical status of local hyperthermia, radiation, and chemotherapy in locally advanced malignancies. AB - The combination of ionizing irradiation and local hyperthermia therapy has been demonstrated to be efficacious in a variety of localized neoplasms. One of the most consistent conclusions from this experience, however, is the finding that large tumor size is a significant negative prognosticator for attaining complete tumor regression. During the past decade investigators have begun to look at the possibility of adding chemotherapy to thermo-radiotherapy in order to improve the efficacy of treatment in patients with large tumors. This review article summarizes the recent clinical experience with such triple modality therapy. PMID- 9329556 TI - Rationale and clinical status of 41.8 degrees C systemic hyperthermia tumor necrosis factor, and melphalan for neoplastic disease. AB - Dramatic clinical results have been obtained in malignant melanoma and sarcoma using hyperthermic limb perfusion in combination with tumor necrosis factor (TNF) and melphalan (L-PAM). In order to extrapolate these results to systemic treatment, a preclinical research program was initiated to study the interactions of hyperthermia, TNF, and L-PAM. Based on these results, a Phase I clinical trial of whole body hyperthermia (WBH) and L-PAM was initiated and completed. Clinical results obtained were consistent with initiating two second generation studies: a) a Phase II study of WBH and L-PAM for malignant melanoma; b) a Phase I study of WBH, TNF and L-PAM. Both of these studies are currently active at the University of Wisconsin Comprehensive Cancer Center. The following review summarizes the laboratory and clinical data obtained to date regarding this systemic multi-modality treatment approach. PMID- 9329558 TI - Systemic hyperthermia and ICE chemotherapy for sarcoma patients: rationale and clinical status. AB - Preclinical studies are consistent with the concept that 41.8 degrees C whole body hyperthermia (WBH) can enhance the therapeutic index of specific chemotherapeutic agents. These laboratory investigations resulted in 2 phase I clinical studies, which also support this hypothesis. These trials were extended to 2 sequential phase II investigations of WBH plus ifosfamide, carboplatin and etoposide (ICE) for refractory sarcoma patients. The first study (involving 12 patients) using extra-corporeal WBH was prematurely closed to adopt a less toxic WBH technology, i.e., the radiant heat Aquatherm. To date, 12 patients have been accrued to the Aquatherm trial. Projections regarding reduced morbidity were correct. The response rate for ICE/WBH is currently 63%. The review to follow will summarize the results of these trials, as well as the laboratory and clinical data which serve to explicate the dramatic clinical results observed to date. PMID- 9329559 TI - Prognostic value of preoperative serum levels of CEA, CA 19-9 and CA 72-4 in gastric carcinoma. AB - We studied the relevance of CEA, CA 19-9, CA 72-4 and the common classical prognostic factors (age, sex, tumor infiltration, N-classification, staging, grading and Lauren classification) in gastric carcinoma. PATIENTS AND METHODS: Survival function estimates were calculated according the method to Kaplan-Meier. The patients were separated into two groups according to preoperative marker levels. Fixing specificity at 100% for healthy people, cut off levels were calculated. Survival curve differences were assessed using the log-rank-test. Multivariate Cox proportional hazards regression analysis was performed. The mantel-Haenszel method was used to assess the 2-year survival rate of patients with gastric carcinoma and high versus low levels of tumor-associated antigens adjusted to tumor stages. The study was performed on the frozen sera (stored at 80 degrees C) of 103 patients with histologically proven gastric carcinoma. RESULTS: The tumor stage (log-rank chi-square = 55.9; P < 0.0001) represents the best prognostic factor besides preoperative values of CA 19-9 (log-rank chi square = 13.9; P < 0.001) and CEA (log-rank chi-square = 12.2; P < 0.001). CA 72.4 shows a log-rank chi-square of 6.9 (P < 0.01). We found no statistically significant correlation between survival and sex, tumor grade and Lauren classification. The importance of different parameters in providing additional prognostic information was evaluated by multivariate analysis. Only patients after curative surgical intervention (n = 55, R0) were considered. Cox proportional hazards regression analysis yielded an adjusted relative risk of 2.4 in patients with a preoperative CEA concentrations > or = 4 ng/mL vs. < 4 ng/mL, of 2.8 in patients with a preoperative CA 19-9 concentration > or = 60 U/mL vs. < CA 19-9 and of 1.8 for stage III/IV vs. stage I/II (P < 0.05). For CEA the 2-year survival rates in the group of patients with preoperative serum concentrations > or = 4 ng/mL versus < 4 ng/mL and stadium III/IV were 14% versus 29% and in stadium I/II 50% versus 83% (P < 0.05). For CA 19-9 the 2-years survival rates in the group of patients with preoperative serum concentrations > or = 60 U/mL versus < 60 U/mL and stadium III/IV were 14% versus 28% and in stadium I/II 40% versus 83% (P < 0.05). CONCLUSION: The postoperative R-classification and the tumor stage represent the best prognostic information besides the preoperative values of CA 19-9 or CEA, respectively. The predictive information provided by preoperative CEA and CA 19-9 serum levels is additional to that obtained from other factors investigated. PMID- 9329560 TI - Pathological aspects of prostate cancer and benign nodular hyperplasia. AB - The aim of the study was to investigate tumor suppressor genes (TSGs) which play a role in the pathogenesis of prostate cancer. Additionally, different prostate tumors were immunohistochemically related to their potential precursor cells, the basal cells and the glandular secretory epithelium, which differ in their hormone responsiveness. By PCR-amplification of microsatellite-DNA we found allelic losses of chromosomal loci near known or putative TSGs to increase with the malignancy grade of prostate carcinoma. When investigated for numerous markers common and endometrioid carcinoma were immunohistochemically related to the secretory epithelium while the rare basal cell tumor, containing the estrogen receptor, squamous cell carcinoma and urothelial carcinoma showed features of the basal cells. In histopathological differential diagnosis high molecular weight basal cell keratins, prostatic acid phosphatase and prostate specific antigen may be of value. Stromal type nodular hyperplasia and the fetal prostate mesenchyme had common immunohistochemical features which might reflect analogous development. PMID- 9329561 TI - Cobas Core CA 19-9 II EIA: new CA 19-9 enzyme immunoassay with high correlation to radioimmunoassays. AB - Especially in patients with pancreatic carcinoma (1-5) (89%) the high molecular weight mucin CA 19-9 (MG 210 kD) shows a high specificity (98.5%) and sensitivity (up to 82%) (6-7). The mayor problem in the field of CA 19-9 diagnostics is the lack of satisfying comparability between CA 19-9 test kits: external proficiency studies (8) indicate a discrepancy between enzymatic and isotopic tests regarding their correlation. The Cobas Core CA 19-9 EIA II contributes to the standardisation of the CA 19-9 kits and allows the switching over from CA 19-9 IRMA's to EIA's. The test can be performed on the random access immunochemistry analyzer Cobas Core with a total assay time of 75 minutes by using 20 microliters of serum or plasma specimens. The analytical sensitivity was determined to be > 0.3 IU/ml. The 2 recalibrators cover a measuring range from 0 to 400 IU/ml. A high-dose-hook effect was not observed up to a concentration of 400,000 IU/ml. Interferences from the sample caused by heterophilic antibodies are reduced to a minimum. Precisions for intra-assay and inter-assay ranged below 5% and 6% respectively. In summary, the Cobas Core CA 19-9 II EIA exhibits a significant improvement regarding the correlation to CA 19-9 radioimmunometric methods (CIS, Centocor). PMID- 9329562 TI - Beta-2-microglobulin--a rapid and automated determination for a broad range of clinical applications. AB - The clinical evaluation of the Cobas Core beta 2-Microglobulin EIA was performed on the random access analyzer Cobas Core. The coefficients of variations for the intra-assay and inter-assay precisions ranged between 3% and 6%. In comparison to healthy persons (0.7-2.7 mg/l), significantly elevated serum levels of beta 2-m were found in patients with lymphoproliferative disorders like monoclonal gammopathies of the IgG (1.98-78 mg/l; p = 0.0001), IgA (1.3-7.1 mg/l; p = 0.0002) and of the IgM (2.1-8.7 mg/l; p = 0.0001) type, in malignant lymphoma (1.5-33.5 mg/l; p = 0.0001) and in chronic lymphatic leukemia (1.5-22.4 mg/l; p = 0.0001). In cases of HIV-infection, increasing levels of beta 2-m exhibited an inverse correlation to the CD4+ T-lymphocyte count (p = 0.0001) and indicated disease progression. In patients having renal transplantation a rejection of the graft was accompanied by a rise of the beta 2-m serum level. In summary, the data clearly indicate the reliability of the measuring system as well as the clinical relevance of beta-2-M determinations for such patient groups. PMID- 9329563 TI - PSA standardization: a review of NCCLS, Stanford and Abbott efforts. AB - The wide variances between prostate specific antigen (PSA) values for various assays and the demonstration that many of these differences are due to calibration differences has resulted in efforts to develop standards for PSA. Two major efforts are underway in the USA. The National Committee for Clinical Laboratory Standards (NCCLS) has published proposed guidelines for the purification and characterization of PSA and PSA-ACT (alpha-1-antichymotrypsin) complexes for primary standards. Furthermore, the Second Stanford PSA Conference proposed a mixture of 90% PSA-ACT and 10% PSA (90:10 standard) with a biochemically defined concentration to calibrate total PSA assays. Significant improvements in agreement between assays was observed with the 90:10 standard as compared to results with kit calibrators. The NCCLS has reviewed and adopted the 90:10 proposal. Thus, biochemically defined standards are preferred over immunoassay defined standards due to differences in assays and laboratory methods. The use of the 90:10 standard will be a major step towards improving the agreement between PSA immunoassays. PMID- 9329564 TI - The soluble interleukin-2 receptor--a marker for squamous cell carcinoma of the upper aerodigestive tract. AB - Various studies indicate that the soluble interleukin-2 receptor (sIL-2R) is of value as marker in chronic and malignant disorders. No data are available on sIL 2R levels in head and neck cancer. The serum of 86 patients with squamous cell carcinoma of the head and neck (SCCHN) prior to any therapy and of 25 healthy controls was taken. Six month later the serum of 49 patients was taken again. sIL 2R levels were determined with ELISA. Patients with SCCHN prior to therapy had significantly elevated sIL-2R serum levels (895 U/ml) compared to controls (437 U/ml). After an eight month follow up 78.4% of the SCCHN patients with tumor recurrence had increased and 80% of the carcinoma patients without recurrence had decreased sIL-2R levels compared to prior to therapy. sIL-2R seems to be an unspecific marker for SCCHN and may be of prognostic value. PMID- 9329565 TI - Ki-67, ploidy and S-phase fraction as prognostic factors in gastric cancer. AB - The prognostic value of the immunohistochemical expression of Ki-67 in paraffin embedded specimens was studied in 242 patients with gastric adenocarcinoma. The results were compared with ploidy and S-phase fraction (SPF) obtained by DNA flow cytometry. Both SPF and ploidy correlated with survival in univariate analysis. In multivariate survival analysis, stage of disease, DNA ploidy and presence of distant metastases emerged as independent prognostic parameters. There was no significant difference in survival between patients having tumours with high and low Ki-67 labelling, neither in univariatenor in multivariate survival analysis. PMID- 9329566 TI - AFP isoforms and their clinical significance (overview). AB - AFP has maintained great clinical relevance as target tumor marker of germ cell tumors (YST = yolk sac tumors) and hepatocellular cancer (HCC) besides its occasional elevation in other malignancies. Its organ and tumor specificity is further limited by elevation in benign liver diseases (BLD) and in pregnancy. The binding of various lectins to its carbohydrate chain (5%) in addition to mere isoelectric focussing has been successfully used in the differentiation of AFP source and disease. The primarily used lectins in affinity electrophoresis and chromatography are Concanavalin A and lens culinaris agglutinin (LCA). The inclusion of other lectins (e.g. erythroagglutinating E-PHA, allomyrina dich. aggl. A lectin) and a more practical nomenclature have been used for characterization of 4 reactive patterns of cord serum/liver/BLD, HCC, g.i. tumor and YST type. In particular, data are reviewed concerning the clinically important liver AFP differentiation by means of LCA and E-PHA differentiation. In conclusion the investigation of AFP lectin binding in cases unexplained AFP elevation may be helpful in the differentiation of AFP source and malignancy. PMID- 9329567 TI - Biliary mucin secreted by cultured human gallbladder epithelial cells carries the epitope of CA 19-9. AB - Serum CA 19-9 is increased in patients with different gastrointestinal malignancies. Unfortunately, CA 19-9 is also detected in high concentrations in normal bile causing unspecific serum elevations during inflammatory disease of the biliary tract and cholestasis. In order to identify the source of CA 19-9 in bile, the capacity of cultured human gallbladder epithelial cells (HGBEC) to secrete CA 19-9 was investigated. Cells were harvested from gallbladders removed by cholecystectomy and cultured for up to 14 days in collagen I coated 24-well culture dishes. CA 19-9 was measured in the culture medium by a solid-phase CA 19 9 EIA (Boehringer). In addition, culture medium was separated by Sepharose 4B-Cl, Concanavalin-A (Con-A) and CA 19-9 affinity chromatography. Significant CA 19-9 activity was measured in the culture medium after a 24 hour incubation period. Following separation of the culture medium by Sepharose 4B-Cl and Con-A affinity chromatography, 90% of the CA 19-9 activity was recovered in the exclusion volume (> 2000 kDa) from which 90% were identified as Con-A negative. A close correlation was found between CA 19-9 and concentrations of mucin purified from human gallbladder bile. Furthermore, CA 19-9 affinity chromatography selectively extracted mucins from the culture medium of HGBEC. Finally, addition of the mucin secretagogue bethanechol (6 mM) to the culture medium increased CA 19-9 activity in the medium. In conclusion, normal HGBEC secrete mucins carrying the epitope of CA 19-9. During inflammatory biliary disease unspecific elevation of CA 19-9 in serum may reflect both inflammatory hypersecretion and leakage of biliary mucins into serum. PMID- 9329568 TI - Preoperative serum levels of CEA and CA 19-9 and their prognostic significance in colorectal carcinoma. AB - The prognostic information provided by preoperative serum CEA, CA 19-9 antigen assays as compared with the classical prognostic factors (age, sex, tumor infiltration, tumor stage (Dukes') and R-classification) in 495 patients with colorectal carcinoma was analysed. PATIENTS AND METHODS: Survival function estimates were calculated according to Kaplan-Meier. The patients were separated into two groups according to the preoperative marker levels. Fixing specificity at 100% for healthy people, cut off levels were calculated. Survival curve differences were assessed using the log-rank-test. Multivariate Cox's proportional hazard regression analysis was performed. The Mantel-Haenszel method was used to assess the survival rate of patients with colorectal carcinoma and high versus low levels of tumor-associated antigens according to tumor stages. The study was performed on the frozen sera (stored at -80 degrees C) of 495 patients with histologically proven colorectal carcinoma. RESULTS: The Dukes' stages (log-rank chi-square = 231.9; P < 0.0001) represent the best prognostic factor besides the preoperative values of CA 19-9 (log-rank chi-square = 162.5). CEA shows a log-rank chi-square of 71.4. Thus, CEA and CA 19-9 can be used to discriminate two groups of patients with significantly different survival times (P < 0.0001). The importance of different parameters in providing additional prognostic information was evaluated by multivariate analysis (Cox's model). Estimated relative risks of death adjusted for tumor stage were 5.5 for Dukes' stage A versus Dukes' stage B/C and Dukes' stage B/C versus Dukes' stage D, respectively and an increasing relative risk of 27.5 for Dukes' stage A versus Dukes' stage D (P < 0.001). The relative risk for preoperative CA 19-9 serum concentrations (> or = 60 U/mL versus < 60 U/mL) was 2.3 (P < 0.001) for preoperative CEA concentrations (> or = 4 ng/mL versus < 4 ng/mL) 1.4 (P < 0.07). For CEA the 2-years survival rates in the group of patients with preoperative serum concentrations > or = 4 ng/mL versus < 4 ng/mL and Dukes' stage D were 16% versus 38%, in Dukes' stage B/C 73% versus 91% and in Dukes' stage A 100% versus 98%. For CA 19-9 the 2-years survival rates in the group of patients with preoperative serum concentrations > or = 60 U/mL versus < 60 U/mL and Dukes' stage D were 10% versus 39%, in Dukes' stage B/C 58% versus 87%. In the group of patients with Dukes' stage A with serum levels > or = 60 U/mL a 2-year survival rate of 100% was found. In the corresponding group only one patient exists. CONCLUSION: The postoperative Dukes' classification provides the best prognostic information besides the preoperative values of CA 19-9. The predictive information provided by the preoperative CA 19-9 serum level is additional to that obtained from the other factors investigated. PMID- 9329569 TI - Tumor marker concentrations in normal and malignant tissues of colorectal cancer patients and their prognostic relevance. AB - Tumor markers CEA, CA19-9, CA15-3, CA125, AFP, beta-HCG, SCC were measured quantitatively in the serum, tumor tissue and healthy colonic mucosa of patients with colorectal cancer. We wanted to investigated whether there is a difference in concentration between patients with and without recurrence of cancer. During the follow-up period 14 of 38 patients showed tumor recurrence. The patients with cancer relapse had higher preoperative serum levels of CEA and CA19-9 and in the histologically normal colonic mucosa they had higher concentrations of CEA, CA19 9, SCC and low CA15-3. The highest values of CEA, CA19-9, and SCC occurred in the mucosa of patients developing local cancer recurrence. Marker concentrations in tumor tissues themselves did not differ between patients with or without tumor relapse. Though confirmation in a larger number of cases is needed we conclude from these results that tumor marker concentrations in the healthy colonic mucosa of patients with colorectal cancer may become valuable indicators of the risk of tumor recurrence. PMID- 9329570 TI - Is the tumor marker CASA dependent on the menstrual cycle? AB - The time course and characteristics of the tumor marker CASA (Cancer Associated Serum Antigen) during the menstrual cycle were analysed. We investigated whether hormonal influences in women and the score of this tumor marker ought to be considered in diagnosis. It could be demonstrated that CASA is not influenced by physiological alterations of the hormonal status in regularly ovulating women. PMID- 9329571 TI - Prognostic significance of CA125 in patients with ovarian cancer and secondary debulking surgery. AB - Prognostic outcome of patients with bulky disease after primary surgery in ovarian cancer remains extremely poor. One possible approach to achieve prolonged survival is secondary debulking surgery, but only in those patients without residual tumor after the second surgery. In 79 patients with secondary debulking surgery preoperative CA125 values were determined. In 52% of the patients with CA125 values below 35 U/ml a tumor free situation could be achieved at secondary debulking. In contrast, the percentage of patients macroscopically free of disease with levels above 35 U/ml was only 22%. Furthermore, there was a significant difference in survival time depending on CA125 values at the time of secondary debulking. Patients with levels below 35 U/ml survived 49 months, women with values above 35 U/ml survived only 30 months respectively. In conclusion, CA125 is an important prognostic tool for predicting a tumor free situation at secondary debulking surgery. In patients with values above 35 U/ml secondary debulking should be indicated restrictively, even if other preoperative diagnostic tools would predict a tumor free situation after secondary cytoreductive surgery. PMID- 9329572 TI - Significance of tumor marker determinations in the primary therapy of ovarian cancer. AB - In 296 patients with primary ovarian cancer the sensitivity and specificity of CA125 and carcinoembryonic antigen (CEA) were determined. High CA125 values can be found in serous cystadenocarcinomas, CA125 can also be detected in patients with mucinous carcinomas. Only in cases negative for CA125 should other tumor markers be determined. As a second choice CA72-4 is proposed, since because of its low sensitivity of only 9% CEA should no longer be regularly determined in ovarian cancer. Preoperatively determined CA125 values show no prognostic significance in primary ovarian cancer. The means of CA125 half life in primary therapy demonstrated prognostic significance, and therefore patients with a worse prognostic outcome can be spared the adverse side effects of ineffective chemotherapy. PMID- 9329573 TI - Mammary serum antigen (MSA), Ca 549, CA 15-3 and CEA in breast cancer preoperative sensitivity and correlation to prognostic factors. AB - Previously a higher preoperative sensitivity of the mucin like glycoprotein Mammary Serum Antigen (MSA) compared to established tumor markers was reported. The aim of our study was the comparison of MSA and CA 549, CA 15-3 and CEA in primary breast cancer and the correlation to prognostic factors. In 119 patients MSA and CA 549 serum levels were analysed by ELISA, CEA and CA 15-3 levels by LIA. We received the following sensitivities: 30.2% for MSA (cut off = 55 U/mL), 21.8% for CA 549 (cut off = 12.6 U/mL), 20.5% for CA 15-3 (cut off = 25 U/ml) and 10.7% for CEA (cut off = 6 ng/ml). Significant correlations were found between MSA concentrations and grading (p = 0.006), tumor size (p = 0.005) and metastases (p = 0.03). No correlations were found to tumor type, hormone receptors, lymph node status and cathepsin D. MSA is a valuable tumor marker with the highest preoperative sensitivity. Further studies on the value of MSA in the follow-up are necessary. PMID- 9329574 TI - Value of Cyfra 21-1, TPA, and SCC-Ag in predicting extracervical disease and prognosis in cervical cancer. AB - BACKGROUND: The value of serum Cyfra 21-1 level was compared with tissue polypeptide antigen (TPA), and squamous cell carcinoma antigen (SCC-Ag) in squamous cell cervical cancer. MATERIALS AND METHODS: Pre- and post-treatment serum levels of patients were compared with lymph node status, parametrial involvement, and prognostic data. RESULTS: Pretreatment serum levels of each marker were significantly related to tumor stage, tumor size and the presence of either lymph node metastases or parametrial involvement. The clinical performance of each marker in predicting extracervical disease at initial diagnosis appeared to be similar. None of these tests achieved the conditions of a perfect test. Pretreatment serum levels of each marker showed prognostic value in the univariate analysis. The clinical performance of post-treatment SCC-Ag levels in predicting complete remission versus the presence of tumor during follow-up was better than Cyfra 21-1 or TPA. CONCLUSION: SCC-Ag appears to be more useful than Cyfra 21-1 for monitoring patients with cervical cancer. PMID- 9329575 TI - Prognostic significance of squamous cell carcinoma (SCC) antigen in primary advanced and recurrent cervical carcinoma. AB - Between January 1986 and June 1992 56 patients with cervical carcinoma were treated with cytostatic drugs in our department. In all patients showing primary response to therapy, the SCC and CEA levels fell rapidly to normal after one or two cycles. In contrast, clinical remission was not obtained in those patients with levels which remained high or rose again following an initial decrease. Using tumor markers, treatment can be individualized so that, cases of therapy failure or further tumor progression can be detected early and the patient can be spared the severe side effects of treatment. PMID- 9329576 TI - Clinical value of CYFRA 8/18 and TPS in the diagnosis and follow up of invasive breast cancer. AB - In a prospective study, we evaluated the diagnostic accuracy of CYFRA 8/18, TPS, CEA and CA 15-3 among 415 patients in various clinical situations of invasive breast cancer and 244 women with benign breast diseases. In comparison to TPS, the sensitivity of CYFRA 8/18 was slightly lower as well in local malignancy (25% vs. 30%) as in metastatic cancer (54% vs. 57%), but in follow up care the rate of false positive results of TPA (> or = 25%) seems to be higher than that of CYFRA 8/18 (> or = 17%). In conclusion, the clinical value of CYFRA 8/18 and TPA appears to be similar. PMID- 9329577 TI - CA 125 is an indicator for pleural metastases in breast cancer. AB - In this study we analyzed the role of CA 125 serum levels in the diagnosis and follow-up of pleural metastases in breast cancer patients. CA 125 serum levels were measured in patients with lung and/or pleural metastases: a) at the time of the detection of the metastases, b) 3 months thereafter during therapy and c) before and after drainage of pleural effusions. In all patients the occurrence of metastases at other localisations was excluded. For CA 125 measurement ELISA sandwich assays were performed. In 76 patients with metastases CA 125 levels were elevated in 8% of patients with lung metastases, in 89% with pleural metastases and in 94% with both sites of metastases. In patients with lung metastases, the CA 125 concentrations did not follow the clinical course of the disease, while in patients with pleural metastases CA 125 levels followed the course of disease in 25 of 25 progressions and in 5 of 6 remissions. After the puncture of pleural effusions the CA 125 levels decreased in 7 of 14 patients and remained stable in the other 7 patients. CA 125, an established marker for diagnosis and follow-up of ovarian cancer, could also be considered as a relatively selective marker for the diagnosis and follow-up of pleural metastases in breast cancer patients. PMID- 9329578 TI - Cyfra 21-1 and TPA as markers in malignant mesothelioma. AB - BACKGROUND: Serum concentrations of CEA and Cyfra were compared in patients with mesothelioma. MATERIALS AND METHODS: Serum levels of patients were analysed for prognosis of survival and follow up of disease. RESULTS: Cyfra 21-1 and TPA have significant prognostic value for survival. CONCLUSION: Assays measuring CK19 fragments seem to be good markers in the management of patients with malignant mesothelioma. PMID- 9329579 TI - Serum S-100 has prognostic significance in malignant melanoma. AB - BACKGROUND: Pretreatment serum values of S-100 and NSE were compared in patients with malignant melanoma. MATERIALS AND METHODS: Pretreatment serum levels of patients with malignant melanoma were analysed for stage of disease and survival. RESULTS: S-100 pretreatment serum values seem highly specific, but have low sensitivity. CONCLUSION: Further evaluation is needed with an assay with improved sensitivity. PMID- 9329580 TI - Screening for prostatic carcinoma with prostate specific antigen. AB - The usefulness of prostate specific antigen (PSA) in screening for prostatic carcinoma was studied in 262 inpatients of the department of internal medicine. All patients underwent a rectal digital examination and determination of PSA by the Tandem-E method (Hybritech). The plan was to perform biopsies if there were suspicious findings on the rectal examination or if the PSA value was more than 10 ng/ml. The PSA values were < or = 4 ng/ml in 219 patients (83.6%), > 4 to 10 ng/ml in 27 patients (10.3%) and > 10 ng/ml in 16 men (6.1%). In consideration of the severity of disease which limited life expectancy we did not perform a biopsy on 37.5% of the patients with PSA > 10 ng/ml. 7 patients with prostatic carcinoma were found. Their PSA values varied between 11.2 and 875 ng/ml. The cancer detection rate was highest for the combination of a suspicious rectal examination and a PSA value > 10 ng/ml (70%). PMID- 9329581 TI - The use of prostate-specific antigen (PSA) for the monitoring of radiation therapy in prostate cancer. AB - BACKGROUND: The classic methods for surveilling the efficacy of radiotherapy in prostate cancer are not accurate enough. The objective of this analysis was to determine whether prostate-specific antigen could perform this task. MATERIALS AND METHODS: From 1/95 to 10/95, 16 patients were treated at our clinic. 7 of these underwent primary irradiation, 4 treatment for local recurrence, and 5 had adjuvant radiotherapy. Radiotherapy was carried out with a total dose of 60 Gy in 30 fractions, 10 fractions per week, to the prostate bed plus 2 cm safety margin. RESULTS: PSA levels usually start to decline between the 3rd and the 4th week of radiotherapy with a half-life of 2.5 months. Five patients had equal or rising PSA levels, including all three patients with recurrent tumor. DISCUSSION: In most cases, PSA declined continuously from the 3rd week of therapy. Our median half-life was similar to other reported results. Persisting or rising PSA levels are an indicator for local or distant recurrence; all our patients who developed a recurrence showed a corresponding PSA increase. PMID- 9329582 TI - The ratio of free to total prostate specific antigen: an advantageous addition in the differential diagnosis of benign hyperplasia and cancer of the prostate? AB - This study examined the clinical relevance of the determination of free PSA (f PSA) in addition to total PSA (t-PSA). PATIENTS AND METHODS: Both total PSA- and free PSA-values of frozen sera obtained pretherapeutically from 80 patients with carcinoma (PC) and 171 patients with benign hyperplasia of the prostate (BPH) were analysed by means of PSA IRMA and FREE PSA IRMA (IMMUNOCORP/IBL). RESULTS: At 95% specificity (true negative test results), a cut-off value of 16.8 [micrograms/L] was obtained for total PSA (9 patients with BPH [5%] were above this value). For this cut-off value we calculated a sensitivity (true positive test results) of 41%. Using the same criteria for the ratio Q = f-PSA:t-PSA a cut off of 0.083 was found again at a specificity of 95%. In a second step only patients with total PSA values below the cut-off level of 16.8 [micrograms/L]) were considered. Of these patients 11 of 160 with BPH (missing values = 1) and 13 of 33 with PC (missing values = 2) were below the above mentioned ratio (Q = 0.083). Considering both steps (total PSA and Q) 46 patients with PC were detected correctly and 20 patients with BPH would have been biopsied unnecessarily (positive biopsy rate: 70%). CONCLUSION: High total PSA levels are a very good indicator for the presence of prostate cancer. There is still concern to improve the differentiation between the diagnosis between BPH and PC, when an intermediate or low value (< or = 95% specificity) is observed. The determination of Q is only useful in this range and might be helpful for the clinician's decision to apply or avoid biopsy. PMID- 9329583 TI - Increased discrimination between benign prostatic hyperplasia and prostate cancer through measurement of percentage free PSA. AB - OBJECTIVE: The value of Prostate-Specific Antigen (PSA) for the early detection of Prostate Cancer (CaP) is controversial due to an appreciable false positive rate causing unnecessary biopsies. As PSA exits in both free and bound forms the percentage of free PSA was found to be lower in CaP than in Benign Prostatic Hyperplasia (BPH). We investigated whether the percentage of free PSA offers better discrimination on the detection of CaP. MATERIAL AND METHODS: In a retrospective analysis the percentage of free PSA was determined in the sera of 50 consecutive patients with histologically proven BPH (n = 30) and clinically localised CaP without metastases (n = 20; pT1-3 No Mo). Serum levels of free PSA and total PSA were determined employing a chemiluminescent enzyme immunoassay. RESULTS: Patients with CaP demonstrated a lower percentage of free PSA (median: 8.5, range: 2.7-24.5) than patients with BPH (median: 22.35, range: 8.9-66.7). (p < 0.001). CONCLUSION: Determination of percentage of free PSA enhances the discrimination between BPH and CaP and may reduce the number of unnecessary biopsies in patients with elevated PSA. PMID- 9329584 TI - Metastatic workup of patients with prostate cancer employing skeletal alkaline phosphatase. AB - PURPOSE: To compare the efficacy of two tests, prostate-specific antigen (PSA) and skeletal alkaline phosphatase (SAP) as staging markers to discriminate patients with cancer of the prostate (CaP) with bony metastases (M1) from those without bony metastases (Mo). MATERIAL AND METHODS: Forty-seven untreated patients with Mo (n = 26) and M1 (n = 21) CaP were entered in this study. Serum concentrations for SAP and PSA were determined using two immunoassays. RESULTS: None of the Mo patients but 65% of the M1 patients exhibited a SAP value above the reference range (< 19 ng/ml). A corresponding cut-offpoint of 100 ng/ml for PSA showed that 27% of Mo patients and only 65% of the M1 patients exhibited a value > 100 ng/ml. This resulted in a sensitivity and specificity of 65% and 100% for SAP and 65% and 73% for PSA. CONCLUSION: Our findings suggest that SAP could become a useful marker in the evaluation of patients with newly diagnosed CaP as it seems to provide additional information concerning the skeletal status of these patients. PMID- 9329585 TI - Can thyroid peroxidase be used as a complementary tumor marker besides thyroglobulin? Preliminary experience with determination of TPO in differentiated thyroid carcinomas. AB - Serum TPO concentrations were studied compared with the hTG levels in 47 patients suffering from different thyroid carcinomas (before and several times after the first, second or third radioiodine therapy respectively. An immunoluminometric coated tube assay was applied recognizing this enzyme (Brahms Diagnostica GmbH). Elevated TPO levels could be seen in 18/47 pts. All of them suffered from large postoperative remnants or metastases. An increase of TPO concentration was seen frequently at days 2 and 3 following the first or second radioidine application. Although the hTG response seems to be more sensitive, TPO may be helpful to reduce the number of false positive hTG findings. PMID- 9329586 TI - Prognostic value of TP53 protein accumulation in human primary breast cancer: an analysis by luminometric immunoassay on 1491 tumor cytosols. AB - The tumor suppressor gene TP53 is implicated in the regulation of normal cell growth and division, DNA repair and apoptosis. Mutations in this gene usually give rise to a conformationally altered protein which is stably expressed at high levels. We have studied TP53 protein accumulation in routinely prepared cytosols from 1491 human primary breast cancer specimens (median follow-up of patients alive, 66 months), using a quantitative luminometric immunoassay (LIA). The TP53 LIA values varied between 0 and 153.53 (median 0.20 ng/mg protein). Median TP53 levels were significantly higher in ER- and PgR-negative tumors. In Cox univariate regression analysis, when analyzed as a continuous variable, increasing TP53 levels were related with a poor relapse-free survival (p < 0.01). In multivariate analysis for relapse-free survival, including age, menopausal status, tumor size, nodal status and steroid hormone receptor status, TP53 accumulation, when analyzed as a dichotomized variable, was an independent factor for predicting the rate of relapse with a relative relapse rate (95% confidence limits) of 1.39 (1.19-1.63). In conclusion, the LIA for the TP53 protein can easily be performed on cytosols routinely prepared for steroid hormone receptor analysis, it is a quantitative assay, and it may be useful in establishing the relation of TP53 accumulation and breast cancer prognosis. PMID- 9329587 TI - Circadian variations of interleukin-2 receptors, serum thymidine kinase and beta 2-microglobulin in patients with non-Hodgkin's lymphoma and normal controls. AB - In this study we investigated whether circadian rhythms of interleukin-2 receptors (sIL-2R), serum thymidine kinase (sTK) and beta-2-microglobulin (beta 2M) are apparent, which may influence tumor marker detection. Blood was drawn every two hours over 24 hours from 6 patients with NHL, 3 healthy donors and three patients with non-hematologic disorders. The serum levels of the three markers were measured. The three normal volunteers showed circadian variations of sIL-2R, sTK and beta 2M best described by a sinusoidal curve. This rhythm could also be demonstrated in three of the six patients with NHL for all three markers. In three patients the circadian rhythm of one marker was disturbed. The three NHL patients with consistent circadian rhythms in all three markers achieved a complete remission, whereas the patients with disturbed rhythm progressed under therapy. We showed that circadian variations of sIL-2R, sTK and beta 2M follow a sinusoidal circadian rhythm in normals and patients with NHL. Disturbance of this rhythm may be related to a worse prognosis. PMID- 9329588 TI - Homogeneous immunoassays using rare earth cryptates and time resolved fluorescence: principles and specific advantages for tumor markers. AB - We have developed a new type of long lived fluorescent tracer, the rare earth cryptates formed by the inclusion of a Eu3+ ion into the intra molecular cavity of a macrobicyclic ligand containing bipyridine units. This tracer is used in a new homogeneous assay format based on spectral and temporal selectivity as well as on an amplification of the cryptate fluorescence. As a consequence of these features, the measurement of the specific signal is totally shielded from media interaction and corrected in real time for sample to sample optical variation. We show the example of a PSA assay where an analytical sensitivity of 0.03 ng/ml is obtained and an AFP assay where measurements in the first minutes of the incubation allow a rapid estimation of the AFP concentration and therefore an immediate dilution of the samples if required. These results illustrate the performance of this new homogeneous method and point out the specific advantages it allows in terms of sensitivity, precision and flexibility. It is therefore particularly well adapted for the assay of analytes like tumor markers where both sensitivity and wide dynamic range are needed. PMID- 9329589 TI - Free PSA in the detection of prostatic carcinoma. AB - A new possibility for improved differentiation between malignant and benign prostatic disease is the determination of free-PSA in the indifferent grey area of total PSA between 2 and 30 ng/ml. In a retrospective study of 106 men with a total PSA between 2 and 30 ng/ml we studied the ratio of free to total PSA. The differentiation between prostatic carcinoma (PCa) and benign prostatic hyperplasia (BPH) was verified by randomised biopsies. PSA was measured with Tandem-E, Hybritech, USA and free-PSA with Tandem-R, Hybritech, USA and Immunite R, DPC-Biermann, USA. Patients (pts.) with an untreated, virgin PCa releaved a highest quotient free-PSA/PSA lower than 0.25. The highest quotient in pts. with treated PCa was 0.51 and in pts with BPH was 0.52. Therapy of PCa with LHRH analogues changed free-PSA toward a BPH-profile. Both kits used for free-PSA gave similar results. Our study suggests, that every free-PSA higher than 25% of PSA should not be a valuable, supplementary parameter for pts. with unclear diagnosis. PMID- 9329590 TI - CA125 based diagnosis and therapy in recurrent ovarian cancer. AB - Approximately 85% of patients with advanced ovarian cancer will experience recurrence of the disease. In 311 patients CA125 serum levels were determined at follow up investigation in our department. A sensitivity of 92% and specificity of 89% were calculated. Reflecting the further course of disease, specificity could be increased up to 97%. Using 25 U/ml/month as the upper limit of grade of CA125 increase, a 100% specificity for detecting recurrence could be achieved. In conclusion, recurrence diagnosis and even therapy should be started in cases of either significantly raised CA125 levels or elevated grade of CA125 increase. PMID- 9329591 TI - Evaluation of serum neural cell adhesion molecule as a prognostic marker in multiple myeloma. AB - Serum neural cell adhesion molecule (NCAM), a possible prognostic marker for multiple myeloma (MM), was determined by means of an enzyme immunoassay, which showed good linearity and high precision. In 95% of healthy controls (n = 70), NCAM values were below 18.7 U/mL. In patients with monoclorlal gammopathies of undetermined significance (MGUS) (n = 31) or polyclonal gammopathies (n = 53) the cut off was 23.1 U/mL. MM in active stage (n = 52) showed significantly higher NCAM levels (p < 0.001) than in asymptomatic stage (n = 44). In active myeloma the sensitivity of serum markers were found to be: NCAM 40%, beta 2-microglobulin beta 2-M) 52% and serum thymidine-kinase (S-TK) 41% (cut off defined on MGUS). The combined sensitivities ranged between 55 and 60% (NCAM+ beta 2-M, beta 2-M+S TK, NCAM+S-TK). No correlation with beta 2-M or S-TK could be demonstrated. However, NCAM values were correlated with the concentration of monoclonal immunoglobulin (IgG-paraprotein: r = 0.45; IgA-paraprotein: r = 0.58). In the follow-up of patients with myeloma, NCAM values decreased in response to chemotherapy and were low in smouldering myeloma. But in three patients with progression NCAM did not reflect the tumor activity. At the time of censor, 80% of patients (n = 80) with a pre-treatment NCAM of < 18.5 U/mL and 61% of patients with a NCAM of > 18.5 U/mL were still alive. NCAM showed a low prognostic significance (log-rank: p < 0.07). Seven of ten myeloma patients with CD56 expression on plasma cell surface, which was examined by flow cytometry, displayed a high concentration of NCAM in serum. All other non-Hodgkin's lymphomas (21 immunocytoma, 27 chronic lymphocytic leukemia, 16 centrocytic/centroblastic-centrocytic lymphoma, 24 high-grade lymphoma) had low NCAM concentrations in serum and did not significantly vary in follow-up. In conclusion, serum NCAM could be a marker for the staging and monitoring of MM. However, it seems, that NCAM did not provide additional prognostic information relating to beta 2-M, S-TK or paraprotein. PMID- 9329592 TI - Serum thymidine kinase in non-Hodgkin lymphomas with special regard to multiple myeloma. AB - S-TK (Serum thymidine kinase) levels were determined by means of a radioenzyme assay (REA). In 95% of healthy controls (n = 97), S-TK values were below 8.5 U/L. In patients with monoclonal gammopathies of undetermined significance (MGUS) (n = 27) or polyclonal gammopathies (n = 45) the cut off was 10.3 U/L respectively 25 U/L. Patients with viral disease (n = 16), especially infections with Epstein Barr virus, Hepatitis-virus and HIV, had elevated S-TK values of up to 215 U/L. In 95 patients with multiple myeloma (MM) and 103 patients with other various non Hodgkin lymphomas (NHL) S-TK levels were investigated. With regard to monoclonal gammopathies, MGUS had lower S-TK than MM patients (p < 0.05) and patients with stage I MM according to Durie and Salmon had S-TK levels significantly lower than those with more advanced stages (p < 0.01). There was a correlation between S-TK and plasma cell labeling index (r = 0.56, p < 0.001). Patients with chronic lymphocytic leukemia showed significantly higher S-TK levels in the RAI stages 3 and 4 than in stages 1 and 2 (p < 0.01). In cases of other malignant NHL in progression sensitivities of S-TK were found to be: immunocytoma 36%, centrocytic/centroblastic-centrocytic lymphoma 54% and high-grade NHL 40% (cut off defined on lymphomas in remission). S-TK levels varied in MM according to the course of disease and response to therapy decreasing at remission and increasing again at relapse. Analogous variations were found in the other NHL. After two years, 83% of patients with a pretreatment S-TK of < 10 U/L and 47% of the patients with a S-TK of > or = 10 U/L were still alive. S-TK proved to be a highly significant prognostic indicator for MM patients (log-rank and Wilcoxon: p < 0.0001). In the other NHL patients with a S-TK level greater than 10 U/L had a median follow-up of only 7 months. NHL patients with lower S-TK levels did not yet reach the median survival time (log-rank and Wilcoxon. p < 0.005). Our results suggest that the determination of S-TK may help to monitor the clinical course of NHL during therapy and predict the prognosis of NHL. PMID- 9329593 TI - Value of the serum levels of the tumor marker TUM2-PK in pancreatic cancer. AB - The monoclonal antibody pyruvate kinase type tumor M2 (TUM2-PK) has been shown to have a high binding capacity to pancreatic cancer. In present study TUM2-PK serum levels were measured in pancreatic cancer and compared with the reference tumor markers CA19-9, CA50, CA72-4 and CEA. Overall 100 patients were included in this study, 64 patients had a histologically confirmed pancreatic carcinoma, 36 patients gastrointestinal cancer (stomach, colon), 666 healthy volunteers served as controls. Measurements were done by enzymimmunoassay. For the healthy blood donors a cut-off value of 22.5 U/ml was evaluated, which corresponds to 95% specificity. In patients with pancreatic cancer the sensitivities of TUM2-PK, CA19-9, CEA, CA72-4 and CA50 were 71%; 68%, 37%, 49% and 63.4% respectively. Linear regression analysis indicated that there was a positive correlation (r = 0.79). According to the results of our study TUM2-PK has comparable sensitivity but higher specificity than the reference tumor marker CA19-9. PMID- 9329594 TI - Diagnostic value of skeletal AP and PSA with respect to skeletal scintigram in patients with prostatic disease. AB - The aim of this study was to compare the diagnostic value of skeletal AP and PSA with the skeletal scintigram in patients with prostatic cancer. PSA and skeletal AP were measured by Tandem-R-Ostase Assay (IRMA) and Tandem-R-PSA-Assay. 64 patients with prostatic cancer, 20 of them in stage D2 were involved. Patients with a prostatic cancer and bone metastases show remarkable increased skeletal AP concentration with a median concentration of 50ng/ml SAP and 95 ng/ml PSA. Patients without bone metastases have a lower median concentration of SAP at 10 ng/ml and PSA concentration of 40 ng/ml which is within the normal range. Six out of 20 patients at stage D2 showed a significant increase of SAP concentration and even of PSA before bone metastases were seen by skeletal scintigraphy. We conclude when skeletal metastases are assumed in patients with prostatic cancer, a combination of skeletal scintigramm and measurement of SAP seem to be of advantage to recognize patients with bone metastases earlier. PMID- 9329595 TI - Development of the Abbott AxSYM Free PSA assay: performance characteristics and preliminary clinical evaluation. AB - The AxSYM Free PSA assay was demonstrated to have good analytical sensitivity and reproducibility. The F/T ratio determinations for 385 men tested during the Prostate Awareness Week who had biopsies due to an elevated total PSA value and/or a suspicious DRE demonstrated that the percentage of free PSA was lower in patients found to have prostate cancer than those that were biopsy negative for the overall group and for all patient categories examined. The optimal strategy for combining PSA values, F/T ratios, DRE and other clinical and diagnostic parameters to improve the early detection of prostate cancer requires additional clinical studies. PMID- 9329596 TI - First experiences with the use of CASA in mammary carcinoma. AB - The expression of the tumor marker CASA (cancer associated serum antigen) was studied in patients with mammary carcinoma; increased values were almost exclusively seen in cases of manifest metastasis. No correlation existed between tumor stage, lymph node involvement and grading. Marker levels were tested by means of enzyme-immune-assay of the firm medac (Hamburg), with a cut-off value set at 4 U/ml. PMID- 9329597 TI - UGP--a tumor marker of gynecologic and breast malignancies? Specificity and sensitivity in pretherapeutic patients and the influence of hormonal substitution on the expression of UGP. AB - Urinary gonadotropin peptide (UGP) is a 10,300 Dalton peptide which is present in the urine of pregnant women, those with trophoblast disease and those with, certain nontrophoblastic malignancies. We examined the efficiency of UGP measurement at differentiating benign from malignant gynecologic and breast diseases. UGP was measured in 1355 spot urine samples from 841 patients (343 samples from 323 healthy women and women with benign gynecologic and breast diseases, 1012 samples from 518 women with gynecologic malignant diseases or breast cancer). Using a cutoff of > 3 fmol UGP/mg urinary creatinine the specificity was 97%. The sensitivity of UGP was calculated from pretherapeutically collected samples (n = 210). The sensitivity of the test for all malignancies was 26% (ovarian malignancy (n = 27) 52%, endometrial cancer (n = 25) 32%, cervical cancer (n = 49) 29%, breast cancer (n = 72) 19%, vulvar cancer and vaginal cancer (n = 12) 17% and for carcinoma in situ of the breast or the cervix (n = 20) 0%). We also found significantly higher UGP values in postmenopausal women than in premenopausal women. Hormonal substitution significantly lowered the UGP values. PMID- 9329598 TI - Is serum tumor marker half-life a guide to prognosis in metastatic nonseminomatous germ cell tumors? AB - BACKGROUND: The value of serum tumor marker kinetics of AFP and HCG assessed by marker half-life (MHL) analysis for diagnosis and in the follow-up of patients with nonseminomatous germ cell tumors (NSGCT) is still debated controversally. The aim of this retrospective study was therefore to investigate the influence of serum MHL during the first two cycles of chemotherapy on the long-term outcome in metastatic NSGCT. MATERIAL AND METHODS: 147 patients with at least 2 abnormal marker values > 7 days after start of chemotherapy were investigated for HL analysis (HL cut off 3.5 days for HCG and 7 days for AFP). HCG and AFP determinations were performed by a double monoclonal IRMA (HCG) and conventional RIA (AFP) developed by our laboratory. RESULTS: According to these cut offs 35/108 patients (32.4%) had a prolonged HCG HL and 41/114 patients (36%) a prolonged AFP HL. Patients with either MHL prolonged had a significantly inferior overall survival (OS; 10 year OS 37% vs. 75%, p = 0.0005) and progression-free survival (PFS; 10 year PFS 29% vs. 69%, p < 0.0001) than those with a prolonged HCG MHL (OS; 10 year OS 36% vs. 65%, p = 0.003; 10 year PFS 28% vs. 56%; p = 0.001) and even more than those with a prolonged AFP MHL (10 year OS 39% vs. 70%, p = 0.02; 10 year PFS 32% vs. 56%, p = 0.01). CONCLUSION: The remarkable prognostic information assessed by MHL analysis in this retrospective study merits further confirmation by a prospective study for its appropriateness in selecting patients for high dose chemotherapy. PMID- 9329599 TI - A new tumour marker assay for ovarian cancer on the OPUS immunoassay system. AB - The human tumour associated cancer antigen CA 125 is a glycoprotein with high molecular weight. The determination of this antigen has been proven to be useful in the monitoring of patients with ovarian cancer. OPUS OV, the tumour marker assay for the ovarian cancer antigen CA 125 is an ELISA that was developed for the family of fully automated random-access analyzers, OPUS, OPUS Plus and OPUS I Magnum. The assay uses a double monoclonal sandwich format (antibodies B27.1 and B43.13, Biomira/Canada). The first antibody is immobilized on the solid phase of the OPUS modules. Sample is automatically added and incubated for 5 minutes. The addition of a solution of the second antibody conjugated to the enzyme alkaline phosphatase leads to the formation of a sandwich complex within 5 minutes. The last step, the addition of the fluorogenic substrate 4-methylumbelliferyl phosphate, serves both as washing procedure and for the development of the fluorescence signal (kinetic measurement). OPUS OV has an assay range from 0-1000 kU/L with a detection limit of 5 kU/L. Within run cv's are 6-8%. A good correlation was found to commercially available kits for the determination of CA 125. We conclude that this new OPUS OV assay is a valid alternative for use in the routine determination of the cancer associated antigen CA 125 and allows more reliable determinations in terms of random access, speed, and ease of operation. PMID- 9329600 TI - Value of HAMA--determination in clinical practice--an overview. AB - HAMAs (human anti mouse antibodies) in serum of patients may be stimulated as an immunologic reaction to the application of animal protein. They are differentiated into heterophilic (species-unspecific) and mouse-specific anti isotypic (often IgG1) or-idiotypic antibodies as well as according to the human immunoglobulin type (IgG or IgM). Besides infrequent clinically immunologic side effects (anaphylactic reaction), their influence on tumor marker measurements predominates in the follow-up of tumor patients by the possible impairment of newly applied monoclonal antibodies. The detection of HAMAs is performed on suspicion of a tumor by self-made or commercial tests, however, there is no consensus standardization of tests, standards and controls. Experiences are reported herein on the occurrence of heterophilic antibodies, reasons of suspicion, detection and removal, comparative determination of HAMAs by two commercially available tests (ImmunoSTRIP HAMA, Enzygnost) and HAMA induction in patients following immunoscintigraphy (OC-125, n = 27). Besides the rare appearance of HAMAs in patients without pretreatment or following biologic therapy, i.v. application of monoclonal antibodies for diagnosis (immunoscintigraphy) or therapy represents the most frequent reason for HAMA development that should always be ruled out in case of anamnestic indication and clinically unexplained tumor marker changes. PMID- 9329601 TI - Significance of tumor markers during the follow-up of women without symptoms after treatment of primary breast cancer. AB - In Berlin, 23-24 February, 1995, during a "consensus-meeting" several German Oncological Societies ratified their guidelines for the follow-up after primary treatment of breast cancer. In women without symptoms and with no anamnestic or clinical signs for relapse or metastases routine X-ray diagnostics (except mammography), bone scintigraphy, ultrasound and other technical imaging procedures and laboratory evaluation including tumor markers are not obligatory. The relevance of continuously rising tumor markers (e.g. CEA, CA 15-3, MCA) without tumor-related symptoms or manifestation of relapse or metastases is not yet sufficiently studied to give exact information for specific treatment selection. Before this can be done, these factors must not only be shown to be prognostic, their therapeutic relevance must be established by controlled clinical trials. Two examples of such ongoing trials are mentioned. Today's follow-up after curative treatment of local or locoregional breast cancer is symptom-orientated. More data must be accumulated eventually showing, which subgroups of patients could profit from the very early detection of a subclinical metastasizing process. So far these data are lacking, and the use of tumor markers to intensively search for very early metastases does not help the patient or the physician. It gives rise only to unanswerable questions, is costly and time consuming and therefore seems useress. PMID- 9329602 TI - CYFRA 21-1 quantity measurement in the urine of patients with carcinoma of the urinary bladder and tract. PMID- 9329603 TI - Prostate specific antigen (PSA) in serum samples of female patients: possible pitfalls in evaluating the analytical sensitivity of ultrasensitive PSA-assays. PMID- 9329604 TI - Prognostic value of serum analyses of S-100 protein beta in malignant melanoma. AB - In the present study serum levels of S-100 protein beta were measured in 643 patients with cutaneous malignant melanoma. An immuno-radiometric assay with three monoclonal antibodies against bovine S-100 protein beta subunit was used. At the time of blood sampling 553 patients were in clinical stage 1, 24 in clinical stage II and 66 in clinical stage III. The overall survival rate was strongly associated with serum levels of S-100 protein. The observed/expected death ratio was markedly increased with increasing levels of S-100 beta (p < < 0.001). Our data strongly suggest that S-100 beta in serum is an independent prognostic marker and may be useful in identifying high-risk cases and monitoring response to therapy in patients with malignant melanoma. PMID- 9329605 TI - Hormonal treatment with sex steroids in women is associated with lower p105 serum concentrations. AB - BACKGROUND: In breast and ovarian cancer patients elevated serum levels of the oncoprotein p105 were detected. We examined the influence of endocrine factors on the p105 serum concentration. MATERIALS AND METHODS: The sera of 115 and non pregnant women were analysed. Of the premenopausal women (n = 77) hormonal contraception was used by 30 women, and of the postmenopausal women (n = 33) there were 15 women using hormonal replacement therapy (HRT). The serum specimens were analysed by ELISA. RESULTS: Women with hormonal contraception had significantly lower serum levels than premenospausal controls (82-138 fmol/ml, median 109 fmol/ml vs. 103-183 fmol/ml, median 130 fmol/ml; P < 0.001). In postmenopausal women with HRT serum levels were significantly lower than in postmenopausal controls (88-136 fmol/ml, median 117 fmol/ml vs. 99-261 fmol/ml, median 148 fmol/ml; P < 0.001). CONCLUSIONS: Sex steroids seem to modulate c-erbB 2 activity. These results are important with regard to clinical studies on p105 oncoprotein in cancer patients. PMID- 9329606 TI - Anti-p53 autoantibodies in hepatitis C virus-infected patients. AB - Mutations in the p53 gene with generation of circulating autoantibodies to p53 protein (anti-p53) have been recently detected in a significant proportion of patients with different malignant diseases. Using ELISA methods we assessed alpha fetoprotein (AFP) and anti-p53 as serological screening parameters for hepatocellular carcinoma (HCC) in 147 consecutive patients with hepatitis C virus related chronic hepatitis. Liver cirrhosis was histologically diagnosed in 58 patients (39,5%) and a HCC confirmed in 7 patients (4.8%). Serum AFP levels were raised above 20 ng/ml in 26/147 patients (17.7%) and above 100 ng/ml in 5/147 patients (3.4%). In 6/7 patients with HCC serum AFP was raised above 20 ng/ml, but only in 3/7 cases above 100 ng/ml. Autoantibodies to p53 protein were detected in 3/7 patients with HCC, but in 0/140 patients without HCC (sensitivity 42.9%, specificity 100%). In conclusion, the presence of anti-p53 was specific for malignancy and independent of AFP status. Overall, the sensitivity of serological screening for HCC in patients with hepatitis C virus-related chronic hepatitis was improved by combining AFP measurements (level > 100 ng/ml) with the detection of anti-p53. PMID- 9329607 TI - Clinical value of squamous cell carcinoma antigen (SCC-A) in Egyptian gynecologic cancer patients. AB - Squamous cell carcinoma (SCC) antigen levels were measured by immunoparticle assay (IMx) in the sera of 32 patients with gynecologic malignancies, 15 with benign diseases of the genital system and 14 normal healthy controls. At a cut off value of 4.8 ng/ml (100% specificity), the rate of SCC antigen elevation was 100% in vulvar and vaginal cancer (n = 5), 90% in ovarian cancer (n = 10), 60.0% in endometrial cancer (n = 10) and 57.2% in cervical cancer (n = 7). The benign disease's group had 80.0% false positivity at the same cut-off value. Serum SCC-A was found to correlate directly with the clinical stage of disease. A sensitivity of 73.3% was obtained at stage I which gives SCC-A a role in screening the high risk population for gynecological cancer. Concerning the histopathologic type of tumor, serum SCCA was highly sensitive in SCC tumors, in ovarian serous cystadenocarcinoma and in patients with recurrent ovarian cancer. PMID- 9329608 TI - The clinical value of cathepsin-D and TNF-alpha in bladder cancer patients. AB - This study included 34 normal healthy controls, 35 patients with urinary tract bilharziasis and 93 bladder cancer patients (62 were operable cases and 31 non operable). Serum tumor necrosis factor alpha (TNF-alpha) was determined using the enzyme immunoassay reagents supplied by Medgenix Diagnostics, Belgium. Cytosol Cathepsin-D was estimated using the immunoradiometric assay supplied by CIS BIO International, France. The results revealed that at 100% and 90% specificities, cytosol Cathepsin-D had 35.7% and 59.5% sensitivity in bladder cancer patients. Serum TNF-alpha showed sensitivity of 17.0% and 55.0% at 100% and 90% specificities in operable bladder cancer patients and 48.0% and 77.0% in non operable cases respectively. Cytosol cathepsin D and TNF-alpha did not show prognostic values like positive correlation with tumor stages, grades or association of tumors with bilharzial ova or lymph node involvement. PMID- 9329609 TI - Multivariate analysis of DNA ploidy, p53, c-erbB-2 proteins, EGFR, and steroid hormone receptors for short-term prognosis in breast cancer. AB - BACKGROUND: Several molecular-genetic alterations in breast cancer, including aneuploidy, aberrant expression of p53, c-erbB-2, and EGFR have been associated with poor prognosis in breast cancer particularly in lymph node negative patients. To determine the importance of molecular-genetic factors relative to more traditional surgical-pathologic prognostic factors, a multivariate analysis was performed particularly in lymph node positive breast cancer cases. METHODS: One hundred fresh samples of primary breast carcinoma have been studied with flow cytometry for DNA ploidy. On the same specimens steroid hormone receptors (ER and PR) were measured in cytosol fraction using Abbott ELIZA assays, c-erbB-2 and EGFR were determined in the tissue homogenate and mutant p53 protein in the nuclear fraction by Oncogene Science ELISA procedures. In addition, information regarding surgical-pathologic features of the tumor was obtained. Multivariate analysis using Cox's proportional hazards model was done to identify variables predictive of poor prognosis. RESULTS: With univariate analysis, tumor size, lymphnode number, p53, c-erbB-2 were predictive of poor short term prognosis. In the multivariate analysis, only c-erbB-2 (P = 0.001) and p53 (P = 0.05) were significant. Subgroup analysis by nodal status yielded significant association of c-erbB-2 (P = 0.001) and p53 (P = 0.04) with lymph node positive breast cancer. CONCLUSIONS: Among molecular-genetic prognostic factors, c-erbB-2 was the most strongly predictive of poor short term prognosis followed by p53 in lymph node positive breast cancer. PMID- 9329610 TI - CA 125 measurement in the follow-up of breast cancer patients. AB - During follow-up examinations for breast cancer the serum tumor marker CA 125 was measured routinely in the Universitatsfrauenklinik Erlangen. We found elevated CA 125 levels in 18 of 510 clinical asymptomatic patients. These patients were followed closely. A second malignant neoplasm was found in six of these patients, two at a curable early stage. We found metastases of breast cancer in nine patients and benign diseases in three patients. PMID- 9329611 TI - Tissue and serum c-erbB-2 and tissue EGFR in breast carcinoma: three years follow up. AB - Amplification of erbB-1 and c-erbB-2 genes has been shown in human breast cancer. Expression of these protooncogenes results in production of epidermal growth factor receptor (EGFR) and c-erbB-2. Both are transmembrane receptors with tyrosine kinase activity. Recent data have indicated that the external domain of c-erbB-2 is shed into the culture supernatant of certain breast cancer cell lines and sera of breast cancer patients. A body of literature has shown that the overexpression of these receptors in malignant tissue and c-erbB-2 when shed into serum is associated with bad prognosis. In the present work, tissue EGFR and c erbB-2 were determined in the membrane fractions of histopathologically verified malignant and normal tissues from the same breast of 94 patients. These values were also determined in 48 tissue specimens of benign mastopathies. Serum c-erbB 2 was quantified in breast cancer patients (n = 105), patients with benign breast disease (n = 48) and 30 apparently healthy women as controls. Patients were followed up by determination of serum c-erbB-2 for one year and clinically for three years to detect any distant metastasis or recurrence. The levels of tissue and serum c-erbB-2 and Estrogen receptors were significantly higher in the carcinomas and sera of breast cancer patients than benign breast diseases or normal controls. Follow-up, although short, of pre-operative serum c-erbB-2 showed a prognostic value (P = 0.007) better than that of tumor size (P = 0.04), EGFR (P = 0.18), nodal involvement (P = 0.25) and tissue c-erbB-2 (P = 0.85). The shedding of soluble fragments of c-erbB-2 into the serum seems to be a characteristic of the potentially malignant cell. The EGFR mean level, however, was significantly lower in malignant tissues than benign and normal ones. A new definition of EGFR status was developed. Accordingly, the recurrence of the disease was more frequent among patients with negative EGFR. The present work did not reveal any correlation between tissue, serum c-erbB-2 or EGFR on one hand and age, menopausal status, stage, histological type and grade of carcinomas and nodal involvement on the other hand. The present work showed an inverse correlation between estrogen receptor level and level of EGFR in malignant tissues. PMID- 9329612 TI - Epidermal growth factor receptors: status and effect on breast cancer patients. AB - The predictive potential of Epidermal Growth Factor Receptor (EGFR) is still a matter of debate. EGFR was quantified biochemically using an enzyme immunoassays malignant and normal tissues from the same breast (n = 94) as well as benign mastopathies (n = 40). The mean level of EGFR in malignant tissues showed a significant decrease from the control and benign ones with a weak positive correlation existing between EGFR level in malignant and control tissue of the same breast. Statistically, no cut-off line could be drawn between malignant and non malignant tumors due to the large overlap in their values. On the contrary to reports on EGFR, when the patients were classified according to the relative changes in EGFR from malignant and adjacent tissue, patients with a relative EGFR decrease (negative EGFR) in malignant tissue showed the poorer prognosis in short term follow up. Mean EGFR values in malignant or normal tissue, or the difference between them, did not show any significant correlation with the age of the patients, menstrual status, clinical stage, type and grade of the carcinoma and lymph node involvement. The present work showed also an inverse correlation between EGFR and Estrogen Receptor (ER) level in the malignant tissues. PMID- 9329613 TI - Computer modeling of cytokeratin release in clinical oncology. AB - The levels of cytokeratins (CK) in serum of cancer patients have been widely used for monitoring progression of cancer growth and the effectiveness of cancer treatment. Previous studies have shown that the release of CK by tumors in patients is a complex process which depends on the rate of cell damage caused by an increasing tumor mass, or by the tumor treatment, but is not in any simple manner correlated to the number of proliferating cells or to the total tumor mass (1). The complexity of the CK-releasing process has been analyzed by a computer model which mimics the progress of tumor growth, allows the introduction of different types of treatment (i.e. irradiation, chemotherapy and surgery), and computes the amount of CK released by the tumor, and the level of CK in blood and blood clearance. The computer model can be used to obtain a better understanding of the interactions of various factors, for scheduling of treatment and CK sampling, and for analyzing the effects of treatment. PMID- 9329614 TI - Cytokeratin profiles in neck cystic lymph node metastases of tonsillary origin. PMID- 9329615 TI - Active-specific immunotherapy of pancreatic carcinoma: usefulness of human pancreatic carcinomas in preparing autologous tumor vaccines. AB - Using a mouse model we investigate whether pancreatic carcinoma cells can serve as a basic material for the preparation of tumour vaccines. Human pancreatic carcinomas grown in nude mice were dissociated and stimulated with interferon gamma and tocopherol acetate. Due to the very homogenous tumour material the yield of vital tumour cells even from small specimens was high enough to produce at least three vaccine doses each. Flow cytometric analyses of stimulated cells showed a significant increase in MHC I presenting cells compared to nonstimulated cells. These preliminary data suggest that beneath the successful application of tumor vaccines to renal carcinoma, melanoma, colon carcinoma and different gynaecological carcinomas pancreatic carcinoma could be a further candidate for this kind of therapy. PMID- 9329616 TI - Tissue polypeptide specific antigen (TPS) detects a specific epitope structure on human cytokeratin. AB - A TPS-reactive protein fragment from a human colon adenocarcinoma cell line was purified to electrophoretic homogeneity. N-terminal sequence analysis of the purified 13 kDa protein fragment demonstrated that the component was a fragment of human cytokeratin 18. The M3 monoclonal antibody (detector antibody in the TPS assay) was applied for screening of an expression library. The M3-reactive phage clones were subcloned and PCR amplified. DNA-sequence analysis revealed it to contain a nucleotide fragment corresponding to human cytokeratin 18. Smaller fragment, engineered by PCR and expressed as fusion proteins, demonstrated that the M3 epitope is localized to human cytokeratin 18, amino acid residues 322-340. PMID- 9329617 TI - Effects of interfering and influencing factors on the analyses of p105 (c-erbB 2/HER-2) oncoprotein fragment in serum. AB - BACKGROUND: The 105 kDa extracellular domain of c-erbB-2 encoded protein p185 is detectable in serum. Elevated p105 serum levels were found in patients with breast and ovarian cancer and in pregnancy. The effects of interfering and influencing factors on the measurement are still unclear. MATERIAL AND METHODS: We used an ELISA based on the monoclonal capture antibody OD-3 and investigated possible interfering factors, like changing transporting and storing conditions of blood and serum samples. In order to evaluate a normal range of p105 serum values we examined 71 healthy women. RESULTS: In our tests we found no influence on the reproducibility of results by changing transporting and storing conditions. We could not find any dependency of p105 serum values on the menstrual cycle or the age of controls. Postmenopausal women showed significantly higher serum values than premenopausal women (p = 0.0127). CONCLUSIONS: Clinical interpretation of female p105 serum values requires comparison with normal ranges when considering the menopause as an influencing factor. PMID- 9329618 TI - Modulation of CA-125 release by inflammatory cytokines in human peritoneal mesothelial and ovarian cancer cells. AB - In the present study the modulatory effects of inflammatory cytokines, interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma), on CA-125 release in established ovarian cancer cell lines and in human peritoneal mesothelial cells (HPMC) both grown as monolayers, were investigated. The purity of mesothelial cell cultures were confirmed by the positivity of the cells for vimentin and cytokeratins 8 and 18, and their negativity for markers CD34 and CD68, thus excluding contamination by endothelial cells and macrophages. The preliminary results of CA-125 measurements in the culture medium clearly indicated differences in the pattern of CA-125 expression and release between normal and malignant cells under the influence of inflammatory cytokines. Furthermore, it seems that normal mesothelial cells play a crucial role as a source of CA-125 found in ascitic fluid or in pleural effusions and possibly even in the serum since secretion of this tumor marker into the culture medium was found to be significantly higher in HPMC than in ovarian cancer cells. PMID- 9329619 TI - int-2 and c-erbB-2 gene amplification detected in 70 frozen human breast carcinomas by quantitative polymerase chain reaction. AB - Gene amplification is a common mechanism of proto-oncogene activation and contributes to tumor progression. Analysis of such genetic alterations is relevant to our understanding of tumor genetics and can provide prognostic information for the patients. A rapid, non-radioactive approach based on qdPCR and fluorescent DNA technique was applied for determination of int-2 and c-erbB2 gene amplification and correlated with other prognostic factors in 70 breast cancer samples. ER and PgR were analysed by immunohistochemistry. The mixed template assay showed 96% concordance between calculated and measured gene copy number. int-2 gene and c-erbB2 amplification were both found in 24% of the tumors. The amplification did not correlate with any of the other prognostic factors. 8% of the tumors showed amplification of both genes without significant correlations to any of the other parameters. The fd-PCR assay is a valuable tool for determination of amplification of int-2 and c-erbB2 genes. Therefore, more detailed information about individual tumour biology and outcome may be acquired by this routine assay and probably provide prognostic impact. PMID- 9329621 TI - Enzymun-Test TG: results from external evaluation. AB - A new non isotopic assay for determination of thyroglobulin has been developed by Boehringer Mannheim. This assay uses a new international reference material for calibration, that is thought to replace the current manufacturer specific standardization. Several aspects of the technical performance of this assay have been evaluated in a multicenter evaluation, including precision profile, lower detection limit, recovery testing and method comparison. Additionally reference ranges were established. The presented assay fulfils clinical and analytical needs and offers for the first time a calibration according to a new international certified reference material. PMID- 9329620 TI - Correlation of the EGF-receptor with cell kinetic and classical prognostic factors in breast cancer. AB - SPECIFIC OBJECTIVE: The Epidermal Growth Factor Receptor (EGFR) is a specific cell membrane receptor that shows homology to the product of the oncogene c-erbB2 in human breast cancer. Growth factors bound to the EGFR are able to stimulate the growth of tumor cells in an autocrine or paracrine manner. Our objective was to examine whether there is a relationship between EGFR, cell kinetic prognostic factors (ploidy, proliferation-antigen Ki67) and classical prognostic factors (hormone receptors, menopausal status, nodal status) in breast cancer. METHODS: EGFR was assayed in tumor tissue of 55 patients with breast cancer using an ELISA, the ploidy-status was evaluated by image analysis and Ki67 was determined by immune histochemistry. Estrogen- (ER) and Progesterone-Receptor (PR) concentrations were quantified using a radioligand assay. RESULTS: There was a significant positive correlation between the EGFR and the cell kinetic prognostic factors: EGFR positive tumors were significantly-more often aneuploid and Ki67 positive. In addition there was an inverse association between EGFR- and ER concentration, but no association between EGFR and PR. The EGFR did not correlate with the nodal and the menopausal status. CONCLUSIONS: Our study revealed associations between EGFR, ER, Ki67 and ploidy. Whether these correlations can help to predict the course of disease, providing further information in addition to the conventional factors (nodal status, steroid hormone receptors etc.) has to be investigated by several years of clinical follow up. PMID- 9329622 TI - Distribution of disialoganglioside GD2-antigen and binding of anti-GD2 antibodies on spheroids of neuroblastoma cell line. AB - The ganglioside GD2 is highly expressed on human tumors of neuroectodermal origin. We investigated by scanning electron microscopy the ganglioside GD2 distribution on the surface of spheroids of the neuroblastoma cell line SK-N-LO. About 50% of cells showed GD2 on the plasma membrane and the distribution of GD2 on most of these cells was heterogeneous, with more GD2 at the contact sites of the cells. The binding kinetics of the chimeric anti-GD2 antibody ch14.18 labelled with I-125 on spheroids (average diameter: 450 microns) was determined by gamma counting. Over 4 hours the antibody concentration was raised substantially but up to 24 hours there was only a very slow further increase. A clustered pattern of bound chimeric anti-GD2 antibody ch14.18 was found on a cross-section of the spheroid by autoradiography. PMID- 9329624 TI - Quantification of tumor type M2 pyruvate kinase (Tu M2-PK) in human carcinomas. AB - Proliferating and tumor cells express a certain isoenzyme of pyruvate kinase, called PK type M2. This isoenzyme can be isolated in an active tetrameric and an inactive dimeric form. We have termed this form tumor type M2-PK. This tumor type pyruvate kinase can be quantified by a specific ELISA in blood sera and tumor homogenates. In this study we have compared 26 normal colon mucosa and colon cancer specimens from the same patients. The total specific pyruvate kinase activity and the amount of the tumour type M2-PK measured by ELISA was increased in the tumor samples compared to the normal colon mucosa of the same patient. In normal colon mucosa the specific PK-activity ranged between 0.21 and 1.25 U/mg protein whereas in colon carcinoma we found activities between 0.99 and 7.08 U/mg. The amount of tumor M2-PK measured by ELISA ranged between 0.82 and 27.10 U/mg protein in normal colon mucosa and between 1.96 and 242.40 U/mg protein in colon carcinoma. The tumor M2-PK content in the serum of 666 healthy blood donors was measured by ELISA and compared to sera from 15 colon carcinoma patients and showed a highly significant difference (Mann-Whitney rank sum test, p < 0.001). The values for the 50%-percentiles (median) of blood donors were 10.8 U/ml and 55.0 U/ml for colon carcinoma. PMID- 9329623 TI - The endothelial marker CD 34 in the assessment of tumour vascularisation in ovarian cancer. AB - OBJECTIVE: Tumour angiogenesis as well as the density of newly formed vessels are of potential prognostic relevance in the assessment of malignant neoplasia. Among other monoclonal antibodies the endothelial marker CD 34 is being increasingly investigated in the assessment of tumour vascularisation, especially in vascular tumours. The aim of this study was to determine the value of CD 34 as an immunohistochemical method to quantify tumour vascularisation in ovarian cancers. METHODS: In a preliminary study 30 solid ovarian cancers were investigated with regard to their CD 34 expression. Paraffine embedded specimens were processed immunohistochemically using the PAP-method, in a dilution of the primary antibody CD 34 of 1:50. Morphological aspects, such as tumour homogenicity and vessel distribution, as well as vessel density were analysed. RESULTS: The primary antibody CD 34 reacted positively with the endothelium of arteries, veins and capillaries, noting a more marked expression in small vessels. Furthermore, enhanced staining of those tumour sections with connective tissue was observed, very likely due to the increased vascular pattern of connective tissue. Non homogenous distribution (e.g. "hot spots") was also seen. Overall, an excellent marking and therefore quantification of tumour vessels was achieved using CD 34. SUMMARY: In this pilot study we were able to demonstrate the ability of CD 34 to mark tumour vessels in solid cancers of the ovary. Whether this marker will be of any prognostic relevance in the future is under investigation in a larger patient cohort at present. PMID- 9329625 TI - Growth inhibition of human papillary thyroid carcinoma cells and multicellular spheroids by anti-EGF-receptor antibody. AB - EGF has been reported to stimulate thyroid cell proliferation. In the present study we investigated the effects of anti-EGF-R-antibody (Mab 4253 both as monolayers and spheroids of an oxyphilic, non iodine metabolizing, papillary thyroid carcinoma cell line (ONCO-DG-1) and roughly characterized their EGF-R. Scatchard analysis with I-125-labeled-EGF demonstrated a low number of 1.5 x 10(4) EGF-R per monolayer cell (KD 4.1 X 10(-10) M) and only 6 x 10(3) EGF-R per spheroids cell (KD 5.0 X 10(-10) M). Already 80% of the binding sites were blocked by only 0.44 microgram/ml Mab 425. Proliferathe activity and EGF-R were found to be regularly distributed throughout the spheroids. Adding Mab 425 to medium containing 1 ng/ml EGF, inhibited the growth of monolayer cells by 15% (1 ng/ml Mab 425) and 28% (10 ngiml Mab 425), measured by the MTT-test. The volume growth of spheroids was inhibited by 10-15% using 2 micrograms/ml Mab 425, whereas their viability (MTT-Test) was almost identical. The results show that the anti-EGF-R-antibody (Mab 425) alone is not effective enough for therapeutical use, but it could be of clinical value in conjugation with radionuclides (e.g. I 131) in order to reach metastases not metabolizing iodine. PMID- 9329626 TI - Quality management and international standardization programs for protein immunoassays. AB - The huge number of new immunoassays for the measurement of specific proteins/tumor markers commercially available at present has made necessary comprehensive measures for the quality management of such laboratory tests. Very important for the comparability of test results are international standardization programs for the establishment of consensus/reference methods as well as for the development of suitable reference materials. Such programs are presently conducted by various international organizations and professional societies. Important characteristics of such reference materials are molecular composition, purity, matrix, additives, storage conditions, stability. Internationally accepted reference materials which have been prepared, calibrated and certified will contribute to the harmonization of results with different test systems, improving their quality and clinical use. PMID- 9329627 TI - Tenascin-C tissue concentration in inflammatory and neoplastic diseases of the colon mucosa. AB - BACKGROUND: Tenascin-C is a gycoprotein of the extracellular matrix with predominantly antiadhesive qualities. In the colon mucosa tenascin-C has been found to be induced in inflammatory and neoplastic diseases by immunohistology. This study aimed at quantitating mucosal tenascin-C induction. MATERIALS AND METHODS: Mucosal tenascin-C concentration was determined by Western blotting quantified by densitometry in fresh frozen specimens of the colon from patients with ulcerative colitis, familial polyposis, and colorectal carcinomas. RESULTS: The tenascin-C concentration in normal mucosa was 2.6 micrograms/mg protein (SD +/- 3.4 micrograms/mg). Colorectal adenomas displayed an equal tissue concentration of 2.8 micrograms/mg protein (SD +/- 2.0 micrograms/mg). In ulcerative colitis statistically significant elevated tissue content of 7.5 micrograms/mg protein (SD +/- 4.7 micrograms/mg) was found. Colorectal carcinomas had a tissue tenascin-C level of 18.0 micrograms/mg protein (SD +/- 14.6 micrograms/mg), which was significantly different from the other groups. CONCLUSIONS: Tenascin-C concentration is elevated in inflammatory and neoplastic diseases of the colorectal mucosa. The distinct increase in the tenascin-C content in colorectal carcinomas in contrast to normal levels in colorectal adenomas reflects an association of tenascin-C induction with malignant disease. PMID- 9329628 TI - Analytical and clinical evaluation of a method to quantify bone alkaline phosphatase, a marker of osteoblastic activity. AB - An immunoradiometric assay (IRMA), involving specific monoclonal antibodies (Ostase, Hybritech) and agarose electrophoresis (Isopal, Beckman), two methods for quantification of serum bone alkaline phosphatase (ALP), a marker for osteoblastic activity, were analytically and clinically compared in 293 patients: 79 with end-stage renal failure treated with hemodialysis and 214 with malignant disease. Acceptable within-assay precision was obtained for the IRMA method: 82.5% of the duplicate determinations had a coefficient of variation (CV) < 5%. Curve fitting characteristics were bad and the sensitivity was better than the one mentioned by the manufacturer. Overall correlation between the two methods was good (r = 0.92), except (a) for low values of bone ALP and (b) in some samples with high total liver ALP activity. Low bone ALP determined with the IRMA (< 5 micrograms/L) was confirmed by electrophoresis (< 22 U/L), but ALP activity determined by electrophoresis to be low (< 22 U/L) was not correlated with the IRMA results. After standardizing our results by computing z-values for bone ALP, delta z (= zostase - zelectrophoresis) was significant correlated with liver ALP activity (r = 0.73, P < 0.0001). We conclude that the IRMA for quantifying bone ALP is acceptable. However, when high values for bone ALP are found with the Ostase method, confirmation by electrophoresis remains mandatory to rule out cross-reactivity with high amounts of liver ALP. PMID- 9329629 TI - Comparison of one first and three second generation methods for the determination of CA 125. AB - The monoclonal antibody, OC125 (Centocor, Inc, Malvern, Pa) was the basis for the first generation, one step immunoradiometric assays (IRMA) to detect the CA 125 glycoprotein. Recently, two step IRMA's were developed, the CA 125 II generation assays. In these new assays the CA 125 capture antibody is the M11 monoclonal antibody coated on a solid phase and the OC125 monoclonal antibody is used as the tracer. We compared analytically and clinically one first generation radioassay, and three second generation assays (two radioassays and one ELISA). The ELISA method showed the best within-assay precision and the best curve fitting characteristics. In the clinical comparison, none of the correlations between the first and the second generation methods really satisfied, however the cut off level of 35 U/ml was confirmed. The four CA 125 assays do not yield equal results. As a consequence, the evolution of CA 125 serum concentration during disease monitoring is not reliable when different determination methods are used consecutively. PMID- 9329630 TI - Cytotoxic effect of immunoconjugate composed of glucose-oxidase coupled to an anti-ganglioside (GD2) antibody on spheroids. AB - As a new treatment protocol for neuroblastoma, the chimeric (human/mouse) antiganglioside GD2 antibody chl4.18 is being clinically tested. To improve the therapeutic effect of the antibody alone, we are currently investigating the cytotoxicity of glucose-oxidase coupled to the antibody chl4.18 on spheroids of the neuroblastoma cell line SK-N-LO. The cytotoxic effect of glucose-oxidase is achieved by the production of hydrogenperoxide (H2O2) and probably by the following reaction of H2O2 with iron to form hydrogen radicals (OH.). The cytotoxicity of glucose-oxidase was measured by two viability tests (MTT and WST 1). After a 4 hour treatment of the spheroids with the immunoconjugate, a reduction of viability to 50% (MTT-test) and 25% (WST 1-test), respectively, was obtained. The difference between the results of these two tests, might be explained by the different measurement protocols. PMID- 9329631 TI - Granulocyte-macrophage colony stimulating factor and interleukin-6 enhanced white blood cell synthesis of leukotrienes in chronic myelogenous leukemia. AB - The effect of Granulocyte-Macrophage, Colony Stimulating Factor (GM-CSF) and Interleukin-6 (IL-6) on leukotriene production by CML white blood cells induced by calcium ionophore (A23187) was investigated and the leukotrienes formed were identified and quantified using high performance liquid chromatography (HPLC). The in vivo levels of IL-6 and LTB4 were determined by enzyme immunoassay reagents, while GM-CSF was measured by enzyme amplified sensitivity immunoassay. Although GM-CSF or IL-6 alone did not stimulate the synthesis of 5-lipoxygenase product, preincubation of the white blood cells of CML with GM-CSF or IL-6 for 30 minutes at 37 degrees C enhanced the ionophore A23187 induced leukotrienes synthesis, thus the CML white blood cell suspension primed with GM-CSF or IL-6 produced 26.6 +/- 2.8 and 18.9 +/- 1.3 pmol LTC4/10(6) cells respectively, and 30.2 +/- 3.6 and 25.5 +/- 2.5 Pmol LTB4/10(6) cells. In contrast minute amount of leukotrienes were produced by the control cells. In vivo levels of GM-CSF, IL-6 and LTB4 were investigated in CML and normal healthy donors, elevated chemotactic B4 was found in plasma from CML (267 +/- 70.4) while the mean value in normal healthy donors was (127 +/- 13.6) pg/ml. The plasma level of GM-CSF was 32.4 +/- 15.7 pg/ml and 10.5 +/- 3.1 pg/ml respectively in CML and normal healthy donors, while the mean value of GM-CSF and IL-6 in normal healthy donors were 6.7 +/- 2.2 and 4.9 +/- 2.4 pg/ml respectively. No significant correlation was observed between the level of LTB4 and the level of GM-CSF or IL-6 in CML. PMID- 9329632 TI - P53 auto-antibodies in the sera of patients with oral squamous cell carcinoma. AB - AIM: To investigate the sera of patients with oral squamous cell carcinoma (OSCC) for the existence of p53 auto-antibodies. MATERIAL AND METHODS: The sera of thirty-nine OSCC-patients were investigated. All samples were from untreated patients with no history of another neoplastic disease. The sera of nine healthy subjects served as controls. RESULTS: P53 auto-antibodies were not detected both in all healthy subjects and in 20.6% OSCC-patients with their disease at an advanced stage. We prove that the extinction rates for p53 auto-antibodies in OSCC are very weak in the majority of investigated sera (50%) and are not related to tumour stage. CONCLUSION: It is unlikely that p53 auto-antibodies can serve as a prognostic marker of OSCC. PMID- 9329633 TI - Human monitoring. PMID- 9329634 TI - Biomonitoring of possible human exposure to environmental genotoxic chemicals: lessons from a study following the wreck of the oil tanker Braer. AB - In January 1993 the oil tanker Braer ran aground in the Shetland Islands, Scotland. Approximately 80,000 tons of crude oil were released. Exceptionally high winds caused extensive pollution and exposure of the local population to crude oil. We describe the study which was immediately set in place to examine the exposed population for evidence of genotoxic exposure. Blood samples were taken and primary DNA damage was measured in the mononuclear cell fraction by the butanol modification of the 32P-postlabelling method. Mutation was measured at the hprt locus in T lymphocytes. No evidence of genotoxicity was obtained for either end point, but nevertheless, we believe that useful lessons were learnt, which should be incorporated into the design of future studies: (1) A rapid response is essential, and even if sufficient funds are not immediately available, it is still worth attempting to obtain samples quickly and use cryopreservation, also to attempt to estimate exposure. (2) Adequate numbers of volunteers must be sought, together with enough controls, not just to allow meaningful analysis but to overcome loss of samples and failure of things to go according to plan. (3) Points concerning laboratory practice include: (i) samples should be coded, (ii) clearly defined and proven protocols should be used, (iii) irreplaceable samples should not be used for method development, (iv) should a problem become apparent during the study, work on such samples should cease immediately until the problem is solved, (v) all critical experimental components should be pretested against a laboratory standard. (4) The study design should include replicate experiments to monitor experimental variability and reproducibility, as well as internal standards and cryopreserved "in house" samples. Care must be taken that samples from any one exposure group are spread between a number of independent experiments and that each experiment includes samples from a number of exposure groups. (5) A computerised data base should be maintained with full details of experimental variables, donor attributes, and raw data so that any contribution of experimental artefacts to "outlier" results can be monitored. (6) Because of the nature of the statistical variation for many environmental genotoxicity end points, only a large-scale study is likely to be capable of yielding useful information. PMID- 9329635 TI - A more comprehensive application of the micronucleus technique for biomonitoring of genetic damage rates in human populations--experiences from the Chernobyl catastrophe. AB - The current method for scoring micronuclei as a measure of genetic damage rate in peripheral blood cells is to enumerate this end point in cytokinesis-blocked binucleated cultured lymphocytes. However, one can expect that, due to chronic exposure to genotoxins or inherent genetic instability, micronuclei may be expressed continually in vivo in dividing cell populations such as the progenitor cell lineages leading to mature lymphocytes or erythrocytes. Consequently, micronuclei may already be expressed in peripheral blood lymphocytes prior to culture. In view of these considerations, we have performed a study in children living in regions of Belarus that are contaminated by radionuclides from the Chernobyl disaster and compared their micronucleus frequency in erythrocytes, nondivided lymphocytes, and cultured cytokinesis-blocked binucleated lymphocytes to that of controls living in noncontaminated areas. Preliminary data presented in this paper indicate a significant two- to fourfold increase in micronucleus expression (P < 0.05) in exposed children relative to controls in erythrocytes or peripheral blood lymphocytes in blood smears as well as in mononuclear and cytokinesis-blocked binucleated lymphocytes in cultures. The measurement of micronuclei in nondivided mononuclear lymphocytes represents chromosomal damage expressed during in vivo divisions. The micronuclei in binucleated cultured cells represent micronuclei expressed ex-vivo and may include micronuclei already present in a cell prior to tissue culture. These preliminary data suggest that a different spectrum and level of damage may be observed in nondivided mononuclear lymphocytes, binucleated lymphocytes, and erythrocytes and that a combination of these approaches may provide a more comprehensive assessment of the extent of genetic damage induced by chronic exposure to radionuclides or other genotoxins in haematopoietic tissue. PMID- 9329636 TI - Biomonitoring study of a group of workers potentially exposed to traffic fumes. AB - Risk assessment of environmental pollutants is concerned with the identification of compounds in the environment that might be hazardous to human health: measuring exposure levels, measuring cellular damage and then estimating the probability of harm occurring. The feasibility of such a comprehensive approach has been explored in this study of two groups of workers, one of which may be occupationally exposed to exhaust fumes. No statistically significant difference in cellular damage, as measured by the lymphocyte micronucleus assay, was found between these two groups of workers, although clear differences in exposure levels to volatile organic compounds were detected. A number of other factors identified in the study did show clear effects on micronucleus frequency. PMID- 9329637 TI - Frequencies of hprt mutant lymphocytes in smokers, non-smokers, and former smokers. AB - Previous work with the autoradiographic mutant lymphocyte assay has provided information about the time-course of development of hprt mutations and the persistence of detectable mutant cells in human subjects following therapeutic exposures to genotoxic agents. These early studies also revealed elevations in frequencies of mutant cells in pretreatment blood samples from patients who were current tobacco smokers, but no information was available on former smokers. In the present study, blood samples were obtained from 21 healthy former tobacco smokers who had quit smoking at least 1 year before sampling, 42 subjects who had never smoked, and 23 tobacco smokers. Plasma from all samples was tested for cotinine, a metabolite of nicotine. Current smokers were categorized as heavy smokers (> or = 10 cigarettes per day, cotinine > or = 90 ng/ml plasma) and light smokers (< 10/day, cotinine < 90 ng/ml). Lymphocytes from the blood samples were isolated, cryopreserved, and later thawed and assayed with the autoradiographic hprt assay. The 21 former tobacco smokers had a mean variant (mutant) frequency (Vf +/- standard error) of 1.97 (+/-0.13) per million evaluatable cells. The Vf of 42 subjects who had never smoked was 1.74 (+/-0.13) x 10(-6), not significantly different from the former smokers. The smokers had Vfs of 8.09 (+/ 0.78) x 10(-6) for 18 heavy smokers and 5.22 (+/-1.02) x 10(-6) for five light smokers. The two categories of smokers had frequencies of mutant cells significantly different from each other, and each was significantly higher than non-smokers and former smokers (P < 0.05). Vfs were significantly correlated with both cotinine concentrations and the number of cigarettes smoked per day, P < 0.001. This study demonstrates the sensitivity of the autoradiographic hprt assay for detecting mutagenic effects related to chronic low-level exposures to genotoxins, and indicates that this assay is more likely to detect the effects of recent rather than past exposures. PMID- 9329638 TI - Comet assay in human biomonitoring studies: reliability, validation, and applications. AB - The comet assay (single-cell gel electrophoresis), which measures DNA strand breaks at the level of single cells, is very easily applied to human lymphocytes, and therefore lends itself to human biomonitoring studies. For the examination of DNA base oxidation (a specific marker of oxidative damage), the assay is modified by including a stage at which the DNA is incubated with a suitable lesion specific endonuclease. Here we report on the reliability and reproducibility of this approach, from the level of comparing results from duplicate gels prepared from the same sample of cells, up to an assessment of the natural intra- and interindividual variability in lymphocyte DNA damage measured in groups of normal, healthy human volunteers. We applied the assay in investigations of human disease and occupational exposure of factory workers. PMID- 9329639 TI - DNA damage in leukocytes and buccal and nasal epithelial cells of individuals exposed to air pollution in Mexico City. AB - There is an increased interest in using biological markers to monitor individuals for possible exposure to environmental toxicants. Test systems which permit the sensitive detection of DNA damage and DNA repair are critically important in such studies. The single cell gel electrophoresis (SCG) assay is a rapid and a sensitive method for the evaluation of DNA damage at the single cell level, providing information on the occurrence of DNA single-strand breaks and alkali labile sites using alkaline conditions. In this study, the differences in the basal level of DNA damage between young adults from the south (exposed principally to high levels of ozone) and young adults from the north (exposed principally to hydrocarbons and particles) of Mexico City were investigated by the SCG assay using three different cell types (leukocytes and nasal and buccal epithelial cells). We found an increased DNA migration in blood leukocytes and nasal cells from individuals who live in the southern part of the city compared to those living in the northern part; however, no differences were observed for buccal epithelial cells. These results show the feasability of using the SCG assay to evaluate DNA damage in different tissues and its great potential for use in the monitoring of humans potentially exposed to genotoxic pollutants. PMID- 9329640 TI - Changes in the repair capacity of blood cells as a biomarker for chronic low-dose exposure to ionizing radiation. AB - The purpose of this study was to examine whether changes in the repair capacity of blood cells could be used as a valuable biomarker for radiation exposure. To characterize the repair kinetics in nonirradiated and irradiated cells we first performed in vitro split dose experiments. DNA damage and DNA repair capacity were analysed using the comet assay. Our results showed that the first in vitro irradiation affects the repair system of the cells, resulting in a decreased repair capacity after the second irradiation. Furthermore, the second irradiation results in a large amount of DNA damage in the blood cells. To test whether the analysis of the DNA repair capacity after in vitro irradiation is also a valuable method for in vivo studies of donors exposed to radiation, we analysed the repair capacity of blood cells of two exposed groups: patients subjected to a radioiodine therapy and chronically irradiated volunteers from the Chernobyl region. Both groups also showed a significantly impaired repair capacity indicating a stress on the hematopoietic system. In addition, in the group of the Ukrainians DNA damage after in vitro irradiation was significantly higher than in a control group. These results lead to the presumption that the repair capacity and the DNA damage after in vitro irradiation might be a very useful biological marker for radiation exposure in population monitoring. PMID- 9329641 TI - The effects of vitamin C supplementation on biomarkers of oxygen radical generated damage in human volunteers with "low" or "high" cholesterol levels. AB - A human volunteer study was conducted to test the effect of vitamin C supplementation on biomarkers of oxygen radical-mediated damage in individuals with a range of serum cholesterol levels. A group of 48 non-smokers, 24 men and 24 women, was selected from a panel of over 100 volunteers to give as wide a range of serum cholesterol levels as possible. None of the volunteers was taking medication to control cholesterol levels and they maintained their normal dietary habits so as not to compromise their cholesterol status. Volunteers were allocated to three groups of 16, each consisting of four males with low cholesterol levels (< 6 mmol/L) matched for age and build with four males with high cholesterol levels (> 6 mmol/L) and eight females matched in the same way. A three-treatment, three-treatment period, cross-over design was adopted to take account of any temporal differences in response. The three treatments given were placebo, 60 mg vitamin C/day (the recommended daily allowance) and 6 g vitamin C/day. Each treatment was given for 14 days with 6 weeks between the treatment periods. All procedures were performed to the standards of Good Clinical Practice. Blood samples were taken at the end of each treatment period. Serum was assayed for cholesterol whilst vitamin C, total antioxidant capacity, lipid peroxidation breakdown products and ras p21 protein levels were measured in plasma. Lymphocytes were examined for DNA damage using the Comet assay and chromosome aberration test. The Comet assay was conducted with and without challenge with hydrogen peroxide and the chromosome aberration test with and without challenge with bleomycin. Vitamin C supplementation caused a statistically significant increase in plasma vitamin C concentrations and total antioxidant capacity but did not affect cholesterol levels or ras p21 protein levels. There was a non-significant dose-related decrease in lipid peroxidation breakdown products with vitamin C supplementation. No effect on DNA damage was observed in the Comet assay, either with or without hydrogen peroxide challenge, or in the chromosome aberration test without bleomycin. However, a statistically significant increase in bleomycin-induced aberrations was found after vitamin C supplementation. This may be due to effects of vitamin C on iron status. Comparison of male and female subjects showed statistically significant differences in plasma vitamin C levels, the antioxidant capacity of the plasma and the number of chromosome aberrations induced by bleomycin challenge of lymphocytes in vitro. The results were the same for both low and high cholesterol subjects. This study provides no evidence of a beneficial effect on any of the biomarkers studied of vitamin C supplementation over a short-term supplementation period of 2 weeks in a population of healthy, non-smoking individuals eating a nutritionally adequate diet. PMID- 9329642 TI - Use of fluorescence in situ hybridization (FISH) to assess effects of smoking, caffeine, and alcohol on aneuploidy load in sperm of healthy men. AB - Aneuploidy is a common cause of poor reproductive outcomes in humans and is associated with severe medical problems in liveborn offspring, yet little is known about its underlying cause. A substantial amount of aneuploidy is known to be contributed by the father through cytogenetically abnormal sperm. The purpose of this cross-sectional, observational study was to investigate the potential contribution of common lifestyle exposures (smoking, caffeine, and alcohol) to the aneuploidy load in sperm from 45 healthy male volunteers 19-35 years of age. Sperm FISH (fluorescence in situ hybridization) was used to determine aneuploidy and diploidy frequencies for chromosomes X, Y and 18 across varying exposure levels of smoking, caffeine, and alcohol. Caffeine was significantly associated with increased frequencies of sperm aneuploidy XX18 and XY18, diploidy XY18-18 and the duplication phenotype YY18-18 controlling for alcohol, smoking and donor age. Alcohol was significantly associated with increased frequencies of sperm aneuploidy XX18, diploidy XY18-18 and the duplication phenotype XX18-18 controlling for caffeine, smoking and donor age. There was a suggestive, but unstable, association between smoking and XX18. Even within our truncated age range, we were able to confirm an increased risk for XX18 aneuploidy with increasing donor age. Sperm FISH proved to be a useful biomarker to detect and compare numerical cytogenetic abnormalities in human sperm cells across differing levels of exposure to smoking, caffeine, and alcohol. PMID- 9329643 TI - Influence of GSTM1 and NAT2 genotypes on placental DNA adducts in an environmentally exposed population. AB - The placenta bulky DNA adducts have been studied in relation to metabolic genotypes for glutathione S-transferase M1 (GSTM1) and N-acetyl transferase 2 (NAT2) in 158 mothers (113 nonsmokers and 45 smokers) living in two regions with different annual average air pollution levels of sulphur dioxide, nitrogen oxides, particulate matter < 10 microns, and polycyclic aromatic hydrocarbons. One region was the district of Teplice as the polluted industrial region with mines and brown coal power plants, and the other was the district of Prachatice, an agricultural region without heavy industry. DNA adduct levels were determined by using a butanol extraction enrichment procedure of 32P-postlabeling. GSTM1 and NAT2 genotypes were studied by using polymerase chain reaction. The total DNA adduct levels included a diagonal radioactive zone (DRZ) and one distinct spot outside DRZ (termed X), which was detected in almost all placenta samples and correlated with DRZ (r = .682; P < .001). We found the total DNA adduct levels 2.12 +/- 1.46 (0.04-7.70) and 1.48 +/- 1.09 (0.11-4.98) adducts per 10(8) nucleotides for Teplice and Prachatice districts, respectively, indicating significant differences between both regions studied (P = .004). Elevated DNA adduct levels were found in smoking mothers (10 or more cigarettes per day) by comparison with nonsmoking mothers (3.21 +/- 1.39 versus 1.32 +/- 0.88 adducts per 10(8) nucleotides; P < .001). Placental DNA adduct levels in smokers correlated with cotinine measured in plasma (r = .432; P = .003). This relation indicates that cigarette smoking could be predominantly responsible for DNA adduct formation in placentas of smoking mothers. DNA adduct levels were evaluated separately for non-smokers (1.50 +/- 1.00 vs. 1.09 +/- 0.66 adducts/10(8) nucleotides for the Teplice and Prachatice districts, respectively; P = .046) and smokers (3.35 +/- 1.47 vs. 2.91 +/- 1.20 adducts/10(8) nucleotides for Teplice and Prachatice districts, respectively; P = .384) to exclude the effect of active cigarette smoking on the district variation. These findings indicate that the effect of the environmental pollution in cigarette smokers is practically overlapped by tobacco exposure. No seasonal variation was observed for DNA adduct levels in the overall population studied and no relation between total DNA adduct levels in placenta and levels of vitamins A, C, and E in venous and cord blood was found. A positive GSTM1 genotype was detected in 78 subjects, while negative GSTM1 genotype was found in 80 subjects. Higher DNA adduct levels were detected in the group with GSTM1-negative genotype by comparison with GSTM1 positive genotype (2.05 +/- 1.30 vs. 1.66 +/- 1.39 adducts/10(8) nucleotides; P = .018). This finding is more pronounced in the Teplice district (2.33 +/- 1.36 vs. 1.88 +/- 1.56 adducts/10(8) nucleotides; P = .053) than for the Prachatice district (1.61 +/- 1.09 vs. 1.36 +/- 1.10 adducts/10(8) nucleotides; P = .248) and for nonsmokers (1.45 +/- 0.82 vs. 1.18 +/- 0.93 adducts/10(8) nucleotides; P = .029) more than for smokers (3.45 +/- 1.14 vs. 2.95 +/- 1.62 adducts/10(8) nucleotides; P = .085). Significant district and seasonal differences were found in subgroups with GSTM1-negative genotype. DNA adduct levels in placentas of the GSTM1-negative subgroup were higher in mothers living in the polluted district of Teplice than in Prachatice (P = .012). The adduct levels in placentas sampled in the summer period were higher than in the winter period in the GSTM1-negative population (P = .006). No effect of the NAT2 genotype on DNA adduct levels was observed. PMID- 9329644 TI - Interactions between genetic predisposition and environmental toxicants for development of lung cancer. AB - Significant interindividual variations in health outcome may be caused by the inheritance of variant polymorphic genes, such as CYP2D6 and CYP2E1 for activation, and GSTM1 and GSTT1 for detoxification of chemicals. However, mechanistic studies linking the inheritance of predisposing genes with genotoxic effects towards cancer have yet to be systematically conducted. We have studied 54 lung cancer patients and 50 matched normal controls, who have been cigarette smokers, to elucidate the role of polymorphic genes in cancer. Our data indicates that the inheritance of unfavorable CYP2D6, CYP2E1, and GSTT1 genes in strongly correlated with the smoking-related lung cancer. For heavy cigarette smokers (> 30 pack-years), the smoking habit is the strongest predictor of lung cancer risk irrespective of the inheritance of unfavorable metabolizing genes. For moderate to light smokers (< 30 pack-years), the genetic predisposition plays an important role for the risk (odds ratio = 3.46; 95% Cl = 0.46-40.2). Using a subgroup of the study population, we observed that cigarette smokers having the defective GST genes have significantly more chromosome aberrations as determined by the fluorescence-in-situ-hybridization (FISH) technique than smokers with the normal GST genes (P < 0.001). In conclusion, our study provides data to indicate that individuals who have inherited unfavorable metabolizing genes have increased body burden of toxicants to cause increased genetic damage and to have increased risk for cancer. Studies like ours can be used to understand the basis for interindividual variations in cancer outcome, to identify high risk individuals and to assess health risk. PMID- 9329645 TI - Factors affecting various biomarkers in untreated lung cancer patients and healthy donors. AB - The purpose of the present communication was to determine in lung cancer patients and healthy donors if there was a possible association between cancer and biomarkers of cytogenetic damage and ras p21 oncoprotein levels, and if various exogenous confounding factors (such as smoking habit) and endogenous ones (age, sex, etc.) could affect these biomarkers. Peripheral blood and plasma were collected from 31 lung cancer patients prior to treatment and 35 healthy donors of a similar socioeconomic status and from the same region in Poland. Chromosomal aberrations (CA), sister chromatid exchanges (SCE), high frequency cells (HFC), and proliferative rate index (PRI) were examined from the blood and ras p21 oncoproteins from the plasma. These parameters were used as biomarkers of genotoxic anomalies. All the biomarkers were examined for their relationship to confounding factors of age, sex, smoking habit, and immediate family cancer history. Results were analyzed by a t-test, analysis of variance (ANOVA), and stepwise multivariate regression analysis. All types of CA (including and excluding gaps), percent aberrant cells, SCE, and ras p21 oncoproteins were statistically significantly higher in cancer patients than in the healthy donors. Although there were smaller numbers of females in the cancer patients group who were older than the males, there was a difference due to sex (gender) with statistically significant increases in females for CA, SCE, and HFC, but there was no increase for ras p21 oncoproteins. Cytogenetic damage was not related to other cancers in the immediate families of the groups. All major CA parameters differed significantly between smokers and non-smokers in the cancer patients group, and SCE and HFC differed in the healthy donors group. Such parameters also showed a significant variability with the number of cigarettes smoked and the years of smoking habit. Multivariate regression analyses showed a significant association between cytogenetic damage, ras p21 oncoproteins, and cancer. In conclusion, cytogenetic damage and ras p21 oncoproteins in this study appear to be biomarkers associated with cancer, but have not been proved causally, and confounding factors such as age, sex (gender), and smoking can have an impact on them. PMID- 9329646 TI - No evidence of microsatellite instability but frequent loss of heterozygosity in primary resected lung cancer. AB - We examined microsatellite instability and loss of heterozygosity (LOH) in primary lung tumors from 93 cancer patients, using 16 microsatellite markers. The cases studied included 87 non-small-cell lung cancers (NSCLC) and six small-cell lung cancers (SCLC). All the patients except two were current or former smokers. The microsatellite markers were all dinucleotide repeat sequences from chromosomal locations 1p, 3p, 5q, 8p, 9p, 10p, 11p, 13q, and 17q. None of the tumors showed microsatellite instability (0/93). In NSCLC, 28% (24/87) of the cases showed LOH in at least one locus, whereas, in SCLC, 67% (4/6) had allelic losses. The frequency of LOH differed between the various cell types of NSCLC. The highest frequency was seen in large cell carcinoma (3/6, 50%) followed by squamous cell carcinoma (16/43, 37%) and adenocarcinoma (5/35, 14%). The most common site of LOH was 3p, where markers D3S1284, D3S659, D3S1289, D3S966, D3S647, and D3S1038 were studied. LOH, studied with 9p markers (D9S126, D9S171, D9S162), was less common. The present results, together with earlier reports, suggest that smoking-related primary lung cancers seldom show microsatellite instability but are characterized by frequent LOH. PMID- 9329647 TI - Mutations, tissue accumulations, and serum levels of p53 in patients with occupational cancers from asbestos and silica exposure. AB - In order to determine the relationship between mutations, tissue accumulations, and serum levels of p53 in occupational cancers, we used denaturing gradient gel electrophoresis and DNA sequencing of exons 5-9 of the p53 gene, immunohistochemical analysis for tissue identification of mutant p53 protein, and enzyme-linked immunosorbent assay for serum levels of mutant p53 protein to examine for such alteration in a cohort of individuals with workplace exposure to asbestos or silica, and resultant lung cancers or mesotheliomas. DNA analysis detected mutations in 5 of 18 (28%) tumors, and tissue accumulations of protein were detected in 7 of 20 (35%) tumors; the agreement between mutational and immunohistochemical analyses was significant (kappa = 0.62, P = 0.002). Serum elevations of protein were detected in 4 of 11 (36%) cases with available serum samples; the agreement between tissue alterations and serum elevations was also significant (kappa = 0.71, P = 0.017). In addition, based on the analysis of banked samples, serum results tended to be consistent over time prior to the diagnosis of disease (positive predictive value = 0.67, negative predictive value = 0.83). These results suggest that serum levels of p53 are reasonably accurate in reflecting tissue alterations in p53 at the gene and/ or protein level and may be early biomarkers of disease risk. PMID- 9329648 TI - Use of twins in search for tumor suppressor genes. AB - A new approach is applied in the mapping of tumor suppressor genes: analysis of loss of heterozygosity (LOH) in concordant tumors of monozygotic and dizygotic twins. The method relies on recognition of genome locations undergoing loss in both twins in a high proportion of the set of all twin pairs examined. The method effectively pinpoints, and excludes, the loci of potential tumor suppressor genes. With the help of a high density linkage map any such candidates can be placed within a narrow region of a chromosome arm and perhaps matched with known genes. The analysis of the Swedish Twin Registry has shown a clear genetic component for breast cancer. We have identified mono- and dizygotic twins concordant for breast cancer and collected the pathology specimens. Tumor and normal tissue was microdissected and microsatellite analysis carried out to test for allelic loss (LOH) in entirely new and putative chromosomal loci in this cancer. It can be calculated that using only six pairs of informative monozygotic twins, a locus can be incriminated with a high probability. Using increasingly dense markers and search for homozygous deletions, it should be possible to map one or more candidates for breast cancer. PMID- 9329649 TI - Monitoring of early human fetal development in women exposed to large doses of chemicals. AB - The toxicological in-patient hospital in Budapest is responsible for the care of chemically poisoned persons from a population of 3 million. A population-based prospective epidemiological study of all pregnant women admitted from 1985 to 1993 was used to evaluate effects of large doses of chemicals on human fetal development. Of 559 self-poisoned pregnant women identified, two died from the poisoning. A total of 213 fetuses were in the first month of their postconception development. Of these, 126 had evaluated pregnancy outcomes: 111 ended in very early loss, 3 ended in clinical miscarriage, and 12 survived to delivery. (In addition 73 pregnancies were terminated and one pregnant woman died.) The 12 liveborn infants had two congenital abnormalities that were probably not related to their mother's self-poisoning. Though based on small numbers, these findings are consistent with an "all-or-nothing" effect of chemical poisoning very early in human gestation. PMID- 9329676 TI - Cerebrovascular pathology in Alzheimer's disease: cause, effect or epiphenomenon? AB - Cerebrovascular pathology abounds in Alzheimer's disease. Changes in the endothelium, disruption of the blood-brain barrier and amyloid deposition in the cerebral blood vessels are almost universal in advanced cases. Do these changes represent the cause, the effect, or the consequences of a common pathogenesis of Alzheimer's disease? This volume addresses some of these issues by presenting new knowledge gained from a diversity of fields. Recognition of the mechanisms involved will open new possibilities for therapeutic trials. PMID- 9329677 TI - Brain microvascular changes in Alzheimer's disease and other dementias. AB - Vasculopathy in Alzheimer's disease (AD) may represent an important pathogenetic factor of this disorder. In the present study, microvasculature was studied by immunohistochemistry using a monoclonal antibody against a vascular heparan sulfate proteoglycan. Vascular changes were consistently observed in AD and included decrease in vascular density, presence of atrophic and coiling vessels, and glomerular loop formations. The laminar and regional distribution of these vascular alterations was correlated with the presence of neurofibrillary tangles. However, vascular changes may also follow neuronal loss. Vascular density may be related to a decrease in brain metabolism. Furthermore, one of the main features of AD is the presence of amyloid deposits within brain parenchyma and blood vessel walls. It is not yet clear whether amyloid components are derived from the blood or the central nervous system. Because AD is clearly heterogeneous, based on clinical and genetic data, evidence for either a brain or peripheral origin is discussed. Microvasculature was also analyzed in other neurodegenerative disorders devoid of amyloid deposits including amyotrophic lateral sclerosis/parkinsonism-dementia complex of Guam and Pick's disease. In conclusion, if vasculopathy in neurodegenerative disorders is not directly involved in pathogenesis, it may act synergistically with other pathogenetic mechanisms including genetic and environmental factors. This aspect of pathology is particularly interesting in view of its accessibility to therapeutic interventions. PMID- 9329678 TI - Electron microscopy of amyloid fibrils and microvessels. AB - Senile plaques and microvessels in the cortices of brains with Alzheimer's disease were examined using serial sections under light and electron microscopes. In addition, observations were carried out by using an immunostaining method. The results obtained were as follows: (1) Every senile, plaque contains some amyloid fibrils, and many senile plaques have degenerated capillaries with amyloid fibrils. (2) Amyloid fibrils without senile plaques continue directly to the capillaries. (3) A considerable number of preamyloids are observed surrounding the capillaries when a methenamine silver stain is used. (4) Alpha 1 antichymotrypsin is found in amyloid fibrils, endothelial cells, and vascular feet or astrocytic processes. Our findings strongly suggest that amyloid fibrils forming senile plaques have a close relationship to the capillaries. Moreover, a protease inhibitor, such as alpha 1-antichymotrypsin, could possibly play an important role in producing the amyloid fibrils. PMID- 9329679 TI - The vasoactivity of A beta peptides. AB - We have demonstrated that freshly solubilized A beta peptides can enhance vasoconstriction by phenylephrine or endothelin of isolated rat aorta. Concentrations of peptide producing these effects (100 nM-1 microM) are much lower than those requiring toxicity to endothelial cells in culture, and effects are immediate, not requiring the prolonged time periods for aggregation necessary in A beta cell culture toxicity experiments. Pre-treatment with SOD diminishes the enhancement of vasoconstriction by A beta peptides, suggesting that the effects are partly mediated via a decrease in the nitric oxide/superoxide ratio. Enhancement of endothelin vasoconstriction is observed with A beta 1-40 and A beta 1-42, but not with A beta 25-35 even at 5 microM, again suggesting the mechanism of A beta vasoactivity is distinct from that of A beta cytotoxicity. These observations raise the possibility that A beta peptides in contact with the cerebrovasculature could result in vasoconstriction, hypoperfusion and oxygen free radical imbalance contributing to the neurodegeneration of AD. PMID- 9329680 TI - Production of neurotoxic factors by brain endothelium in Alzheimer's disease. AB - The cerebral vasculature is central to the maintenance of the neuronal microenvironment. We have previously demonstrated that brain microvessels in Alzheimer's disease produce high, potentially toxic, levels of nitric oxide. It is our hypothesis that neuronal injury in Alzheimer's disease occurs because an abnormal endothelium secretes factors that are toxic to neurons. In this study, we report that inhibition of protein kinase C in endothelial cells causes release of a factor that is toxic to neurons in vitro. Our results demonstrate that this endothelium-derived toxic factor is soluble, heat-labile, susceptible to proteolysis, and loses activity with repeated freeze-thawing. The molecular weight of this putative protein is between 10 and 50 kDa, and 8 hours are required after protein kinase C inhibition to detect the endothelium-derived toxic factor in the media. Finally, the endothelium-derived toxic factor kills neurons within 2 hours, suggesting that cell death occurs via necrosis, not apoptosis. These data support the notion that endothelial cells can create an injurious microenvironment for neurons by producing molecules with noxious properties. Altered/dysfunctional endothelial cells in the cerebral microcirculation could be a novel, unexplored source of neurotoxic factors in Alzheimer's disease. PMID- 9329681 TI - Cerebrovascular hypoperfusion: a risk factor for Alzheimer's disease? Animal model and postmortem human studies. AB - Although cognitive impairment during aging is usually associated with neuronal alterations, the cerebrovascular system undergoes prominent alterations in aging as well. Using electron microscopy we previously showed a progressive deterioration of the capillary wall in the cerebral cortex of aged rats. In aged rats the capillary basement membrane (BM) is thickened, massive bundles of collagen fibrils are deposited within the BM, and pericytes are degenerating. A compromized cerebral circulation (e.g., in rats with chronic hypertension) is characterized by an increased number of capillary alterations. In autopsy material (gray matter, gyrus cinguli) of carefully diagnosed patient groups (controls, AD, Lewy body disease, MID and demented Lewy body disease patients) we observed significantly more morphological changes in the capillary bed of demented versus non-demented patients. In both animal and human material morphological evidence points to a relation between energy-dependent nutrient transport across the blood-brain barrier and the ultrastructural deviations. In the AD cases we did not find a correlation between the stage of the disease (Braak I-VI) and the incidence of capillary aberrations, which indicates that the capillary alterations are not a consequence of AD pathology. Simultaneously, we are conducting animal model studies to determine the effects of cerebral hypoperfusion in the rat. Permanent bilateral occlusion of the carotid arteries shifts the behavioral profile of the rats (Morris maze, open field) towards that of aged rats, while the sensitivity for muscarinic ligand agents is altered. PMID- 9329682 TI - Hemodynamic consequences of deformed microvessels in the brain in Alzheimer's disease. AB - The cause of sporadic Alzheimer's disease (AD) remains a mystery. Mounting clinical and experimental data, however, suggest that a cerebral hemodynamic role may affect neuronoglial metabolism. Light and electron microscopy have consistently revealed that the microvasculature in AD brains contains structurally deformed capillaries which create a distorted intraluminal conduit for blood flow. The cerebral capillary distortions can create "disturbed" rather than "laminar" blood flow. Chronically disturbed capillary blood flow will impair normal delivery of essential nutrients to brain neurons as well as impede catabolic outflow of CNS waste products. This condition will negatively affect cerebral metabolism, primarily because of impaired glucose delivery to neurons. Impaired glucose delivery to AD brain results in a patho-chemical cascade that will impair the Na+, K(+)-ATPase ion pump and affect the syntheses of ATP, acetylcholine, and other neurotransmitters. The outcome of this metabolic dysfunction can promote neurofibrillary tangle and senile plaque formation in AD brain. PMID- 9329683 TI - Cognitive impairment and cellular/vascular changes in the cerebral white matter. AB - A possible relation between cerebral white-matter injury and dementia was intuitively attributed by Alzheimer to changes affecting the small penetrating vessels that supply the cerebral white matter. Several observations support the view that white-matter changes detectable by neuroimaging may contribute to cognitive deficits in the elderly. But many questions concerning this matter remain partially answered. In this communication we review: (1) Selected anatomic features of the blood vessels supplying the white matter; (2) possible pathogenetic mechanisms responsible for the white-matter changes; (3) observations on humans and animals suggesting a causal relationship between ischemia/hypoxemia and white-matter injury; (4) epidemiologic studies linking white-matter abnormalities with cognitive disorders. We conclude that abnormalities in the small vessels caused by aging and arterial hypertension, or other processes (cerebral amyloid angiopathy, CADASIL) together with systemic circulatory disturbances, such as abrupt variations in blood pressure values or cardiac diseases, may be the substrate of selective white-matter injury. The damage is structurally characterized by incomplete infarction or selective cellular injury. PMID- 9329684 TI - Cerebral microvascular alterations in aging, leukoaraiosis, and Alzheimer's disease. AB - We have been using alkaline phosphatase (AP) histochemical staining, formerly a research tool for the study of cerebral cortical vascular morphology, to examine pathological changes in the cortex and deep cerebral structures. Deep structures stain similarly to the cortex. The AP stain is found in the afferent vessels (small arteries, arterioles, and capillaries), but not in venules and veins. The stain is also present in leaky vessels, such as those in the area postrema. The vascular supply to the cerebrum is not homogeneous. Supply to the deep white matter, for instance, derives from the leptomeningeal border zone, and then medullary arterioles must wind their way for up to 4 cm before arriving at their ultimate destination. Adding to the difficulties, tortuosities develop in some of these vessels with aging. According to some calculations, hypertensive levels of blood pressure would be required to maintain irrigation through some of these vessels. We have identified a venous alteration that attends aging: periventricular venous collagenosis (PVC) is a previously unrecognized, noninflammatory, mural disease of the periventricular veins. In severe cases, examples can be found of veins that are completely occluded by this process. PVC is found in 65% of subjects over 60 years old, and it strongly correlates with leukoaraiosis. In addition to previously mentioned aging-related changes, we have found extreme tortuosity, multiplications, and aneurysms of the smallest arterioles and lumpy-bumpy capillaries in the deep structures of patients with Alzheimer's disease. PMID- 9329685 TI - Can deposition of amyloid be prevented in Alzheimer's disease? AB - Amyloid is a generic term referring to extracellular fibrillar protein deposits defined by their unique tinctorial, ultrastructural, and protein conformational properties. At least 17 different forms have been identified. In each form the deposit consists of a disease-specific (or pathologic process-specific) protein and a set of common components. The disease-specific protein serves as the basis for the classification of the amyloids. In inflammation-associated (AA) amyloid it can be demonstrated that interactive processes between serum amyloid A (SAA), the AA precursor, and the common components, are likely responsible for AA amyloid deposition. Understanding the details of these interactions provide targets for therapeutic interference that are successful in vivo. Analogous interactions take place between the common components and the beta-protein and beta-protein precursor responsible for the congophilic angiopathy and neuritic plaque amyloid in Alzheimer's disease. Interference with beta-protein/common component interaction in vitro both prevents and reverses beta-protein fibril assembly, indicating that successful delivery of effective agents across the blood-brain barrier should prevent and possibly reverse amyloid deposition in Alzheimer's disease. PMID- 9329686 TI - Coronary artery disease, hypertension, ApoE, and cholesterol: a link to Alzheimer's disease? AB - The premature presence of senile plaques (SP) in coronary artery disease (CAD), and neurofibrillary tangles (NFT) as well as SP in hypertension (HyperT), suggest a neuropathologic link between CAD, HyperT, and AD. Previous MI, CAD and HyperT often occur in and may increase the risk of AD. Expression of Apo-E4 likely increases risk of CAD by elevating blood cholesterol and the risk of AD via proposed interactions with beta-amyloid and/or free radicals (FRs). Any Apo-E4 effect is vague, but FRs probably mediate vascular damage in HyperT. Increasing FR content in the blood is related to increasing CAD severity, while the severity of elevated FR level correlates with how deep into a blood vessel there is activation of the FR scavenger enzyme, superoxide dismutase (SOD). The ApoE genotype and SP/NFT areal densities were determined in a large population of non demented CAD, HyperT and non-heart disease (non-HD) control subjects, and compared to findings in a similar number of AD patients. ApoE immunoreactivity was determined in many individuals. Cholesterol content in cortex was determined by HPLC in a small, loosely age-matched group of Apo-E4 genotype-matched AD, CAD and non-HD subjects. SOD immunoreactivity was also assessed in a number of subjects. The Apo-E4 genotype frequency was increased in CAD, HyperT and AD compared to non-HD controls. Dose of Apo-E4 correlated with SP densities, but not NFT, and only in the non-demented groups. Essentially all SP in CAD, HyperT and non-HD subjects were ApoE-immunoreactive. Cortical cholesterol was increased in CAD and AD compared to controls. SOD immunoreactivity was similar in HyperT and AD; SP were immunodecorated in both. AD, CAD and HyperT may be linked, while CAD and HyperT subjects may die of heart disease before showing cognitive change. PMID- 9329687 TI - Vascular basement membrane pathology and Alzheimer's disease. AB - We have previously demonstrated that the capillary vascular basement membrane (VBM) is pathologically altered in Alzheimer's disease (AD). This microangiopathy is highlighted by the immunocytochemical localization of the three principal intrinsic VBM components: heparan sulfate proteoglycan, collagen type IV, and laminin. These three VBM components also immunolable amyloid deposits and senile plaque-associated glial processes. The present study examines the ultrastructure of the VBM in one brain region severely affected (temporal gyrus) and one relatively spared (cerebellum) from the lesions of AD in both AD and neurological control cases. The cross-sectional area as well as the width of the VBM were found to be greater in AD cortical capillaries. In addition, we found ultrastructural evidence for the activation of microglial-related perivascular cells, and their apparent extravasation through the VBM, findings consistent with the hypothesis that these cells are being recruited as part of a disease-related immune response. The recruitment of these "resting" microglial-like cells from their intra-VBM location to plaques and tangles in AD may explain (1) the thickening and vacuolization of the VBM; (2) the specificity of this VBM alteration to brain regions where there are plaques and tangles; and (3) the source of some of the large number of activated microglia in these affected areas. Thus, while VBM alterations may not be specific to AD, these changes appear to be specifically related to the disease process. PMID- 9329688 TI - Amyloid angiopathy and blood-brain barrier changes in Alzheimer's disease. AB - Evidence is accumulating that suggests that increased permeability of the BBB to blood-borne proteins is favorable for the development of neuropathologic changes such as amyloid angiopathy and formation of amyloid plaques in the AD brain. To study this problem, we applied a quantitative immunocytochemical procedure that enables evaluation of the barrier function of brain microvasculature to endogenous albumin. This procedure was successfully used on scrapie-infected mice, which represent a unique animal model enabling study of an interrelation between BBB function and deposition of amyloid within vascular wall and neuritic plaques. Biopsy specimens obtained during neurosurgical procedures (tumors and dementia) were also examined. Our observations indicate that (1) the vast majority of brain microvessels in scrapie-infected mice and in demented individuals show normal features of the BBB; (2) only those microvascular segments directly surrounded by amyloid plaques or representing amyloid angiopathy show increased permeability to endogenous albumin; (3) numerous immunosignals over the amyloid deposits in plaques and in the wall of angiopathic vessels suggest the affinity of extravasated albumin to the amyloid material. PMID- 9329689 TI - Serum protein leakage in Alzheimer's disease revisited. AB - Leakage of serum proteins into the brain parenchyma has been repeatedly used as evidence of blood-brain barrier (BBB) damage in experimental and human studies. However, there is no consensus in the literature concerning this phenomenon in Alzheimer's disease (AD). We have examined this question by comparing frontal lobe sections in seven groups of patients: Multi-infarct dementia (n = 6), AD with (n = 10) and without (n = 10) infarcts, age-matched controls with (n = 10) and without (n = 10) infarcts, controls with neurodegenerative diseases other than AD, and young controls (n = 10). An additional series compared prospectively followed patients with a diagnosis of either multi-infarct dementia (n = 5) or AD (n = 4). Albumin was detected in white-matter astrocytes in all cases, without significant variation in intensity. In addition, diverse combinations of neurons, astrocytes, and (in AD patients) senile plaques were present in the cerebral cortex in an inconsistent manner. Semiquantitative analysis showed no statistically significant differences among groups. Anti-IgG labeled astrocytes in infarcts only. Complement C3c component was detected in rare amyloid plaques in a minority (15%) of AD cases. Selective labeling of AD-specific lesions in a patchy manner was observed for serum amyloid P. We conclude that there is no immunohistochemical evidence of alteration of the BBB in Alzheimer's disease with or without vascular factors or in old age. Serum amyloid P binds avidly to AD lesions, but our findings are consistent with leakage through the BBB during the agonal or immediate postmortem period. Finally, no specific pattern of abnormality in the BBB was detected in multi-infarct dementia. PMID- 9329690 TI - Interactions of beta-amyloids with the blood-brain barrier. AB - Blood-borne beta-amyloids (A beta s) could affect brain function by (1) crossing the BBB to directly interact with brain tissues or (2) altering BBB function by interacting with the brain capillaries that make up the BBB. Several radioactively labeled A beta s have been examined for such interactions. Blood borne A beta 1-28 is hindered from accumulating in brain by a slow rate of passage across the BBB and by robust enzymatic degradation. A beta 1-40, but not A beta 40-1 or A beta 1-42, is sequestered by brain capillaries, raising the possibility that it could affect BBB function. Small amounts of circulating A beta 1-40 are recovered intact from CSF and brain. A beta 1-40 is degraded by aluminum-sensitive, calcium-dependent intracellular enzymes. Apo-J, which can bind A beta, has been shown with an in situ method to be transported by a saturable system across the BBB. However, our recent work has shown that this system is not operable in vivo, probably because the transporter is saturated at physiological blood levels. In conclusion, A beta s have been shown to interact with and to cross the BBB. PMID- 9329691 TI - ApoE, Alzheimer's disease, and recovery from brain stress. AB - Apolipoprotein E (APOE, gene; ApoE, protein) is the major genetic susceptibility locus for the common forms of Alzheimer's disease (AD). There are three common polymorphisms in the population: epsilon 2, epsilon 3, and epsilon 4. The inheritance of each dose of epsilon 4 increases the risk and lowers the age of onset distribution for AD; epsilon 2 lowers the risk and increases the age of onset distribution. APOE-epsilon 4 has a high positive predictive value for AD, and is clinically useful as an adjunct in the early diagnosis of cognitively impaired patients. The APOE alleles have also been associated with risk of AD with head injury, intraneuronal localization of ApoE in animal stroke models, recovery of function after intracerebral hemorrhage, and recovery of psychological parameters after general cardiac anesthesia. A multifunctional role of ApoE in the brain implicates isoform-specific differences in interactions with several brain proteins including A beta, tau, and MAP-2. Intraneuronal ApoE is increased temporally and in relevant neurons in AD as a function of APOE genotype. Decreased glucose metabolism can be demonstrated by PET imaging in subjects two decades before the median age of onset as a function of APOE genotype. ApoE isoforms may also have different effects as antioxidants. The risk of stroke and vascular dementia has not been confirmed in neuropathological series to be related to specific APOE genotypes. PMID- 9329693 TI - Brain energy metabolizing enzymes in Alzheimer's disease: alpha-ketoglutarate dehydrogenase complex and cytochrome oxidase. AB - PET observations of reduced cerebral glucose metabolism in AD could be explained by a defect in key energy metabolizing enzymes. In particular, levels of two enzymes, cytochrome oxidase (CO) and alpha-ketoglutarate dehydrogenase complex (alpha KGDHC) are generally assumed to be reliably reduced in postmortem brain of patients with AD. How strong is the evidence that brain CO and alpha KGDHC are reduced in AD? In our study CO activity and alpha KGDHC activity and protein subunit levels were measured in cerebral cortex of 19-29 AD patients and 29 control subjects. We found that mean CO activity in cerebral cortex was reduced by 16-26% in the AD group but with almost complete overlap between control and patient ranges. Since our publication in 1992, mean brain CO activity in AD was modestly reduced in 9 independent studies (p < 0.05 in 5). Activity of alpha KGDHC varied widely in control/AD subjects and is not useful as an enzyme marker. Cerebral cortical protein levels of E1-3 subunits, which showed much less variance, were reduced by 23-41% but with large overlap between control/patient groups. We concluded that decreased (i.e., below normal) brain CO and alpha KGDHC is a feature of some, but not all patients with AD. The possible causes and significance of the enzyme changes are discussed. PMID- 9329692 TI - Notch3 mutations in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a mendelian condition causing stroke and vascular dementia. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited condition whose key features include recurrent subcortical ischemic events, migraine attacks and vascular dementia in association with diffuse white-matter abnormalities seen on neuroimaging. Pathologic examination shows multiple small deep cerebral infarcts, a leukoencephalopathy and a nonatherosclerotic nonamyloid angiopathy involving mainly the media of small cerebral arteries. To progress in understanding the pathophysiological mechanisms of this condition, we undertook the identification of the mutated gene. We mapped the CADASIL gene on chromosome 19p13.1. More than 120 families have been referred to our lab. Genetic linkage analysis of 33 of these families allowed us to reduce the size of the genetic interval to less than 1 cM and to demonstrate the genetic homogeneity of this condition. In the absence of any candidate gene, we undertook positional cloning of this gene. We identified, within the CADASIL critical region, the human Notch3 gene, whose sequence analysis revealed deleterious mutations in CADASIL families co segregating with the affected phenotype. These data establish that this gene causes CADASIL. Identification of the CADASIL gene will provide a valuable diagnostic tool for clinicians and could be used to estimate the prevalence of this underdiagnosed condition. It should help in the understanding of pathophysiological mechanisms of CADASIL and vascular dementia. PMID- 9329694 TI - Pathogenesis of decreased glucose turnover and oxidative phosphorylation in ischemic and trauma-induced dementia of the Alzheimer type. AB - The pathogenetic mechanisms causing a dementing brain disease after temporary ischemia, heat shock, or brain trauma are surveyed. These lesions increase beta amyloid precursor protein (beta APP) synthesis. This process is potentiated by an ischemic glutamate release that opens cellular Ca2+ channels, inhibiting glucose turnover and ATP production, which is, under these conditions, accompanied by the generation of beta amyloid (beta A), even in young persons. Beta amyloid starts a vicious circle by inactivating the glycolytic key enzyme, phosphofructokinase, which, with age, exhausts the functional reserve capacity of the brain. This demonstrates that beta A is an epiphenomenon of a dementing brain disease, triggered by the disturbance of glucose turnover and oxidative phosphorylation. Clinical studies have shown that a dementing brain disease can be clearly objectified and monitored by 18F-2-deoxyglucose PET studies. This paper looks briefly at pharmacologic approaches to this disease using models of temporary ischemia, the testing of 14C-deoxyglucose turnover, or examination with 31P magnetic resonance spectroscopy techniques. In conclusion, the key process of all dementing brain diseases of the Alzheimer type is a decreased glucose turnover and subsequently decreased oxidative phosphorylation, linked directly to a secondary amyloid formation and nerve cell atrophy. PMID- 9329695 TI - Neuroimaging of vessel amyloid in Alzheimer's disease. AB - Despite extensive recent advances in understanding Alzheimer's disease (AD) we are unable to noninvasively establish a definite diagnosis during life and cannot monitor the cerebral deposition of amyloid beta protein (A beta) in living patients. We evaluated the use of 10H3, a monoclonal antibody Fab targeting A beta protein 1-28 labeled with Tc-99m. Six subjects with probable AD were studied using single-photon emission computed tomography (SPECT) at times from 0-24 hours following injection. Curves of radioactivity in blood demonstrate a half-life of the injected Fab of 2-3 hours. Images show uptake around the head in the scalp or bone marrow in all subjects. There is no evidence of cerebral uptake of the antibody. Scalp biopsies in all six patients demonstrate diffuse staining with 10H3 of the scalp, a pattern indistinguishable from that found in controls. Evidence of amyloid deposition in the scalp in AD is not seen with other anti-A beta antibodies, suggesting that 10H3 is cross-reacting with another protein. Further studies with anti-A beta antibodies will require longer-lived radionuclides to detect cerebral uptake at later times after injection to allow for complete clearance from the blood. Alternately, imaging using labeled A beta itself may provide a means for noninvasive targeting of cerebral amyloid. PMID- 9329696 TI - Decreased brain glucose metabolism in microvessels from patients with Alzheimer's disease. AB - We studied brain glucose metabolism in patients with Alzheimer's disease and age matched controls in vivo by PET and assessed brain glucose utilization and the phosphorylation constant K3 for hexokinase. In addition we determined in vitro the binding of 2DG and measured its phosphorylation to 2DG-phosphate in cerebral microvessels obtained at autopsy from subjects with Alzheimer's disease and age matched controls. In patients with Alzheimer's disease we found a marked decrease in the kinetic constant K3 for the hexokinase, and a marked decrease in the overall metabolism of glucose in our PET studies; in microvessels there was a marked decrease in the affinity of 2DG and a decrease in hexokinase activity. Alzheimer's disease may be related to a complex alteration in brain glucose metabolism. PMID- 9329697 TI - Clinical studies of cerebral blood flow in Alzheimer's disease. AB - Studies of Alzheimer's disease using single-photon emission computed tomography (SPECT) and positron emission tomography (PET) have found reductions in blood flow and glucose metabolism in temporal and parietal cortex. In 50 AD patients who underwent neuropsychological testing and SPECT perfusion imaging, we found significant correlations between perfusion and performance on the Mini-Mental Status Examination in the frontal and parietal lobes. In addition, specific correlations between perfusion in the frontal lobes and performance on tests of frontal lobe ability were noted. These findings, while suggesting the importance of perfusion measures in determining clinical features of the disease, do not clearly define perfusion changes as primary, since similar findings have been seen when metabolism is studied. In a separate group of 5 AD patients and 16 controls, we used PET with the perfusion tracer HIPDM and examined cerebrovascular reactivity to carbon dioxide inhalation. We found that in multiple brain regions, including the temporal lobes, AD patients showed robust and significant increases in perfusion in response to carbon dioxide that did not differ from the response seen in the controls. Taken together, these results show that while perfusion changes are important in AD, they are not clearly either primary or limiting. PMID- 9329698 TI - Cerebrovascular degeneration is related to amyloid-beta protein deposition in Alzheimer's disease. AB - Current evidence is not inconsistent with the suggestion that cerebrovascular functions decline during normal aging with pronounced effects in both sporadic and familial Alzheimer's disease (AD). The primary causes of these changes remain unknown. It is possible that amyloid beta (A beta) protein is involved in the degeneration of both the larger penetrating vessels as well as the cerebral capillaries that represent the blood-brain barrier (BBB). A beta-induced endothelial changes could also alter muscular tone, resulting not only in increased expression of vascular amyloid precursor protein (APP) and production of A beta, but also in oxidative injury. We used immunochemical methods to examine the status of the perfusing cerebral vessels and the microvascular endothelium in relation to deposition of A beta in AD and non-AD aging control subjects. Double-immunostaining with antibodies to vascular markers revealed marked loss of smooth muscle in larger vessels and absence or attenuation of the endothelium in capillary profiles that still appeared to retain their basement membranes. These vascular changes were predominantly restricted to neocortical regions abundant in A beta deposits. Quantitative studies showed that the microvascular abnormalities were correlated to A beta deposition rather than neurofibrillary tangles or neuronal numbers. Our studies suggest that A beta, irrespective of its origin within vascular myocytes or brain parenchyma, is responsible not only for cerebral amyloid angiopathy, but also for the degeneration of the cerebral microvasculature, which may profoundly affect brain perfusion and BBB functions. PMID- 9329699 TI - Misery perfusion with preserved vascular reactivity in Alzheimer's disease. AB - To elucidate the hemodynamic pathophysiology underlying Alzheimer's disease (AD), cerebral blood flow (CBF), cerebral metabolic rate of oxygen (CMRO2) and oxygen extraction fraction (OEF) were measured with positron emission tomography in 10 patients with probable AD and in 20 age-matched normal volunteers. By the 15O intravenous bolus injection method, CBF was measured during resting state, CO2 inhalation (hypercapnia) and hyperventilation (hypocapnia), and the vascular reactivity (VR) was estimated by comparing the CBF changes (delta CBF%/PaCO2 mmHg) in the hyper- or hypocapnic to the resting state. By the 15O2 single-breath method or 15O steady-state method, CMRO2 and OEF were measured during resting state. Based on 26 regions of interest, local CBF, CMRO2 and OEF were compared statistically between the two groups. As compared with the control group, the mean CBF and CMRO2 decreased to as low as 77.0% and 88.4% of the normal values, respectively, while the mean OEF increased by 12.1% (p < 0.05) in AD patients. These changes were most pronounced in the supramarginal and superior temporal gyri. There was no focal change in VR in the AD group, and no significant difference was seen in VR to either hyper- or hypocapnia between AD and control groups. The results may suggest a vascular involvement, possibly at the capillary level, that might cause a relative misery perfusion syndrome accompanied by preserved vascular reactivity in AD. PMID- 9329700 TI - Magnetic resonance spectroscopic changes in Alzheimer's disease. AB - In vitro and in vivo 31P magnetic resonance (MR) spectroscopy studies of Alzheimer's disease (AD) brain have revealed alterations in membrane phospholipid metabolism and high-energy phosphate metabolism. Mildly demented AD patients compared with control subjects have increased levels of phosphomonoesters, decreased levels of phosphocreatine and probably adenosine diphosphate and an increased oxidative metabolic rate. As the dementia worsens, levels of phosphomonoesters decrease and levels of phosphocreatine and adenosine di phosphate increase. The changes in oxidative metabolic rate suggest that the AD brain is under energetic stress. The phosphomonoester findings support our in vitro findings and implicate basic defects in membrane metabolism in AD brain. MR spectroscopy provides new diagnostic insights and a noninvasive method to follow the progression of the disease and the metabolic response to therapeutic interventions. PMID- 9329701 TI - Clinical trials in dementia with propentofylline. AB - The mode of action of propentofylline (a xanthine derivative) suggested that it would have beneficial effects in patients with Alzheimer's disease or vascular dementia. In four double-blind, placebo-controlled, randomized studies, 901 patients with mild to moderate Alzheimer's disease and 359 patients with mild to moderate vascular dementia were treated for up to 12 months (daily dose of propentofylline: 3 x 300 mg taken 1 hr before food). Patients were assessed at regular intervals for efficacy and safety of the drug. Efficacy variables covered cognitive and global functions as well as activities of daily living. Propentofylline showed statistically significant, clinically relevant improvements over placebo in efficacy assessments, both in patients with Alzheimer's disease and in patients with vascular dementia. The drug was also well tolerated. It had no significant effects on laboratory findings and the adverse events that were considered to be related to the study medication were mostly minor, transient, and affected the digestive and nervous systems. PMID- 9329702 TI - Combined tacrine and estrogen replacement therapy in patients with Alzheimer's disease. AB - In light of evidence that estrogen replacement might affect cholinergic function, we examined possible effects of estrogen replacement therapy (ERT) on clinical response to the cholinesterase inhibitor tacrine in women with Alzheimer's disease (AD). In a previously reported 30-week, randomized, double-blind, placebo controlled, multicenter clinical trial, 14.5% of 318 women with evaluable data had been receiving ERT before randomization. They were randomly assigned to receive placebo or tacrine. Women receiving ERT who were randomized to tacrine improved more than women not receiving ERT who were randomized either to tacrine or to placebo as assessed by cognitive (p < 0.01) and clinical global (p = 0.02) tests. These results provide evidence that prior and continuing ERT may enhance response to tacrine in women with AD. Furthermore, among women on ERT receiving tacrine, there tended to be greater improvement relative to placebo among those without an APOE-epsilon 4 allele. Randomized trials are needed. PMID- 9329703 TI - Omental transposition to the brain for Alzheimer's disease. AB - Omental transposition (OT) to revascularize the brain was first performed in animals in the late 1960s, where it was shown that blood vessels originating from the omentum crossed through the omental-cerebral interface prior to developing into large-sized vessels that penetrated directly and deeply into the underlying brain. The additional cerebral blood flow coming from the omentum was of sufficient volume to protect an animal's brain from cerebral infarction even in the presence of middle cerebral artery ligation. It was also learned that the omentum was a rich source for neurotransmitters and omentum-derived nerve growth substance. OT to the brain is now being done for a variety of conditions which include strokes, TIAs, epilepsy, and Parkinson's disease. Of recent interest is the published information that OT may play some role in Alzheimer's disease (AD). The placement of the omentum on an AD brain has led to a profound decrease in senile plaque formation. The omentum may ultimately prove to be beneficial in reversing or at least stabilizing the dementia associated with the devastation of AD. PMID- 9329704 TI - Support of homeostatic glial cell signaling: a novel therapeutic approach by propentofylline. AB - A pathological glial cell activation, which forces microglia to transform into immunocompetent cells with cytotoxic properties and astrocytes to "de differentiate," presumably adds to neurodegenerative diseases. We examined the modulatory effect of adenosine on the Ca2+ and cAMP-dependent regulation of such reactive glial cell properties in culture and tested possibilities of pharmacologic reinforcement. A strengthening of the cAMP-signaling, as could be achieved by adenosine agonists via a Ca(2+)-dependent action, favored the differentiation of proliferating astrocytes and associated neuroprotective properties (ion homeostasis, formation of trophic factors). But potentially neurotoxic properties of microglial cells were inhibited. Adenosine depressed their proliferation rate and transformation into macrophages, their particularly high formation of reactive oxygen intermediates and the release of the cytokine TNF-alpha. Similar effects were obtained with propentofylline, which acts as selective cAMP/cGMP phosphodiesterase inhibitor and also increases the effective concentration of adenosine by blocking its cellular reuptake. The recently observed induction of microglial apoptosis by elevated extracellular adenosine levels may further contribute to limit secondary nerve cell damage related to a pathological glial cell activation. PMID- 9329705 TI - "Amyloid is not a tombstone"--a summation. The primary role for cerebrovascular and CSF dynamics as factors in Alzheimer's disease (AD): DMSO, fluorocarbon oxygen carriers, thyroid hormonal, and other suggested therapeutic measures. PMID- 9329706 TI - Cerebrovascular changes associated with interleukin-1 beta (IL-1 beta) and histamine (HA) levels in Alzheimer's disease. PMID- 9329707 TI - Causes and consequences of neuronal energy deficit in sporadic Alzheimer's disease. PMID- 9329708 TI - Inherent abnormalities in oxidative metabolism in Alzheimer's disease: interaction with vascular abnormalities. AB - Extensive studies over the last 20 years have documented the existence of inherent abnormalities in oxidative/energy metabolism in Alzheimer's disease (AD). These abnormalities can be linked to characteristics of AD by plausible pathophysiological mechanisms for which there is abundant, robust evidence. The inherent abnormalities in cerebral metabolism of oxygen and glucose can reasonably be expected to interact synergistically with vascular compromise of cerebral oxygen and glucose metabolism in causing brain damage in AD. PMID- 9329709 TI - Microembolic brain injuries from cardiac surgery: are they seeds of future Alzheimer's disease? PMID- 9329710 TI - Amyloid-beta protein angiopathies masquerading as Alzheimer's disease? AB - Current evidence from genetic and epidemiological studies supports the view that Alzheimer's disease (AD) is a heterogeneous disorder. While the disease is pathologically defined by the presence of specified lesions in form of amyloid plaques and neurofibrillary tangles within the parenchyma, other features of pathology are often either neglected or considered coincidental. Our studies suggest that cerebrovascular pathology is inherently part of the disorder, which could be an important factor in a cause or effect manner. We have recently identified subjects having died with severe amyloid beta (A beta) protein cerebral amyloid angiopathy (CAA) in the absence of a profound Alzheimer pathology. These subjects, diagnosed with dementia had a late onset disease and were found at autopsy to exhibit severe CAA but paucity of typical AD changes. Immunocytochemical studies showed numerous microvascular abnormalities as well as characteristic degeneration of the vascular smooth muscle in both surface and intracortical vessels. The pathology was also characterized by occasional intracerebral hemorrhages and multiple infarcts. Further assessment of the abnormalities and amyloid infiltrated cerebral vessels with antibodies to the carboxyl terminus of A beta indicated that the longer, more pathogenic form of A beta(1-42) was found to be highly associated with intracerebral hemorrhages. Our observations suggest that these mild AD cases with a predominantly vascular pathology are variants of AD and bear resemblance to the familial Dutch and Flemish versions of cerebral amyloidosis. We propose that AD is a group of diseases with a variable pathology analogous to the prion diseases, in which a vascular variant also exists. PMID- 9329711 TI - Global Deterioration Scale-related brain hemodynamics and histamine levels in Alzheimer's disease and vascular dementia. PMID- 9329712 TI - Amyloid beta peptide and precursor protein (APP) in mild and severe brain ischemia. PMID- 9329713 TI - Dynamic FDG-PET study in probable Alzheimer's disease. PMID- 9329714 TI - Reduced cortical vasodilatory response to stimulation of the nucleus basalis of Meynert in the aged rat and evidence for a control of the cerebral circulation. AB - In earlier studies we showed that electrical stimulation of the rat nucleus basalis of Meynert (NBM) induces large increases in cerebral blood flow, mainly through cholinergic mechanisms. We then investigated the effect of aging on this influence by measuring cortical blood flow (CoBF) and tissue gas partial pressures (PtO2, PtCO2) in the conscious young adult and aged rat. NBM stimulation increased frontal (+101%) and parietal (+29%) CoBF in young rats. The effects were halved in aged rats. Moreover, PtO2 was significantly increased in young but not in aged rats. By contrast, the corticovascular reactivity to hypercapnia did not differ between young and aged rats, nor did the potentiating vasodilator effect of physostigmine. In combined autoradiographic measurements of cerebral blood flow and cerebral glucose utilization, we recently found that the cortical circulatory response to NBM stimulation was not accompanied by significant metabolic change. Thus, the blood flow changes observed in the cortex cannot be ascribed to increased metabolic activity. The distribution of this uncoupling coincides with that of cholinergic NBM projections directly impinging on cortical microvessels. These data support the cortical microcirculation and suggest the possible involvement of NBM dysfunction in the pathology of cortical microcirculation. PMID- 9329715 TI - Effects of cholinesterase inhibitors on the secretion of beta-amyloid precursor protein in cell cultures. AB - One of the main characteristics of Alzheimer's disease (AD) is the cerebrovascular deposition of the amyloid beta-peptide (A beta), which is derived from a larger beta-amyloid precursor protein (beta APP). The majority of beta APP is processed by either a secretory of lysosomal/endosomal pathway. Carboxyl truncated soluble derivatives of beta APP (sAPP) are generated by the proteolytic processing of full-length beta APP by either alpha- or beta-secretase enzyme. Our objective is to determine whether the processing of beta APP can be regulated by cholinesterase inhibitors, some of which were shown to produce a moderate improvement in memory and cognitive functions in patients with Alzheimer's disease. Here we have analyzed the levels of sAPP derivatives in cultured cells treated with different drugs by immunoblotting samples of conditioned media. The immunoreactive protein bands were developed by probing with the monoclonal antibody 22C11. Treating neuroblastoma, pheochromocytoma and fibroblast cells with high dose of either 3,4-diaminopyridine, metrifonate, or physostigmine did not inhibit the secretion of sAPP. Treating glioblastoma with either 3,4 diaminopyridine or metrifonate showed an increase in secretion of sAPP. However, treatment of cells with tacrine reduced release of sAPP in conditioned media of cell lines studied. The difference in action of metrifonate, physostigmine, and tacrine on beta APP is independent of their anticholinesterase activities. Our results suggests that noncatalytic functions of cholinesterase inhibitors can be utilized to alter the metabolism of beta APP, which might in turn affect the process of deposition of A beta, a key component of the cerebrovascular amyloid detected in AD. PMID- 9329716 TI - The risk of dementia among persons with diabetes mellitus: a population-based cohort study. PMID- 9329717 TI - Changes of microvessels in the brain with Alzheimer's disease. PMID- 9329718 TI - Cerebral hypoxia and ischemia in the pathogenesis of dementia after stroke. PMID- 9329719 TI - Cerebral microischemia as a potential precipitant of the neurodegenerative cascade of Alzheimer's disease. PMID- 9329720 TI - Long-term venous A beta 1-40 infusion in rats causes lung hemorrhage and brain perivascular gliosis. PMID- 9329721 TI - beta-Amyloid-induced cerebrovascular endothelial dysfunction. AB - Cerebrovascular effects of beta-amyloid were investigated using bovine mid cerebral arteries. beta-amyloid-induced endothelial damage was evidenced by increased vasoconstriction, diminished vasodilation and was evident on electron microscopy. The endothelial dysfunction was mediated by reactive oxygen radicals. Vascular damage by beta-amyloid may be an early event in the development of the pathology of Alzheimer's disease. PMID- 9329722 TI - APOE epsilon 4 allele frequency in Alzheimer's disease and vascular dementia in the Spanish population. PMID- 9329723 TI - Inhibition of glutamine synthetase induces critical energy threshold for neuronal survival. PMID- 9329724 TI - beta-Amyloid peptides and enhancement of vasoconstriction by endothelin-1. PMID- 9329725 TI - Amyloid deposition in cerebrovascular angiopathy. AB - Recent evidence suggests that AD is associated with the development of cerebral amyloid angiopathy and with significant changes in the blood-brain barrier glucose transporter and basement membrane protein alterations. It is likely that these changes significantly contribute to chronic cerebral hypoperfusion, possibly resulting in progressive neural death in AD. To investigate the etiology of these changes, we have analyzed postmortem tissue sections of frontal cortex of moderately demented AD cases, stained using both single-label and double-label immunocytochemistry to detect vessels and beta-amyloid. The resultant color images are analyzed using HSV (hue-saturation-value) color image analysis, shape analysis, and histogram analysis techniques. These analyses let us segment the tissue images for further statistical analyses. PMID- 9329726 TI - Tacrine treatment in Alzheimer's disease enhances cerebral blood flow and mental status and decreases caregiver suffering. PMID- 9329727 TI - beta-Amyloid (A beta 1-40)-evoked changes in vascular reactivity are mediated via an endothelium-specific mechanism: studies using rabbit isolated aorta. PMID- 9329728 TI - Impaired dynamic morphology of cerebellar mitochondria in physiological aging and Alzheimer's disease. PMID- 9329729 TI - Human aging: risk factors for cerebral atrophy. PMID- 9329730 TI - Misclassification of dementia subtype using the Hachinski Ischemic Score: results of a meta-analysis of patients with pathologically verified dementias. PMID- 9329731 TI - Types of cerebrovascular lesions associated with severe cerebral amyloid angiopathy in Alzheimer's disease. PMID- 9329732 TI - Chronic ischemia: memory impairment and neural pathology in the rat. PMID- 9329733 TI - The effect of stepwise cerebral hypoperfusion on energy metabolism and amyloid precursor protein (APP) in cerebral cortex and hippocampus in the adult rat. AB - To study the relationship between cerebral blood flow, energy metabolism, and the formation of amyloid precursor protein (APP), an in vivo animal model was established in which stepwise long-term cerebral hypoperfusion states were induced. Adult rats underwent a stepwise chronic cerebral hypoperfusion by crosswise occlusion of the carotid and vertebral arteries with different periods and severity of hypoperfusion until the final steady-state experiment. Investigations of metabolic compounds were done in hippocampus and parietotemporal cerebral cortex. The analysis of energy-rich phosphates and adenosine was examined by HPLC analysis. Substrate concentrations of pyruvate and lactate were measured spectrophotometrically. The APP holoprotein was investigated by immunblot technique. Long-term cerebral hypoperfusion induced a decrease of energy-rich phosphates in the brain areas studied, whereas the concentration of adenosine and the ATP turnover were increased. Pyruvate decreased, and lactate was increased, pointing to a shift in the cytoplasmatic redox state. More severe changes were found in parietotemporal cerebral cortex in comparison to the hippocampus. After 2-vessel occlusion, the concentration of APP decreased, whereas the APP concentration was significantly increased in rat brain after 4-vessel occlusion. It has been demonstrated for the first time in vivo that the reduction in cerebral energy metabolism alters the formation of APP due to cerebral hypoperfusion. PMID- 9329734 TI - Altered choroid plexus basement membrane and epithelium in late-onset Alzheimer's disease: an ultrastructural study. PMID- 9329735 TI - Accumulation of aluminum and silicon in lipofuscin granules suggests retardation of blood-brain barrier function by aging. PMID- 9329736 TI - Altered phosphoglycerides and PLA2 activity in aging mouse brain microvessel membrane. PMID- 9329737 TI - Importance of vascular changes in selective neurodegeneration with thiamine deficiency. AB - These results demonstrate that early alterations in the BBB may underlie selective vulnerability in this model of chronic reduced oxidative metabolism. Changes in the BBB (IgG extravasation) precede alterations in APP processing and cell death. Since thiamine-dependent enzymes are also reduced in the brain in Alzheimer's disease, similar processes may be important in the pathophysiology of the disease. PMID- 9329738 TI - Progress in population analyses of the insulin resistance syndrome. AB - Insulin resistance is associated with a variety of cardiovascular risk factors including hypertension, dyslipidemia, and non-insulin-dependent diabetes. In blacks, the relation between insulin resistance, hypertension, and atherosclerosis has been questioned. Most data collected on the Insulin Resistance Syndrome have been collected in nondiabetic subjects; therefore, no inference can be drawn to exogenous insulin use in diabetic subjects where improved glycemic control is usually associated with improved cardiovascular risk factors (especially dyslipidemia) in the absence of weight gain. PMID- 9329739 TI - Hypertension and the insulin resistance syndrome of rats. Are they related? PMID- 9329740 TI - Obesity genes and insulin resistance syndrome. PMID- 9329741 TI - Development of obesity and insulin resistance in the Israeli sand rat (Psammomys obesus). Does leptin play a role? AB - The Israeli Sand Rat (Psammomys obesus) is an excellent polygenic model for the study of obesity and diabetes. The metabolic characteristics and the heterogeneous development of these defects, including elevated leptin levels, mimic those found in susceptible human populations. Interestingly, only animals that develop metabolic abnormalities demonstrate hyperleptinemia and, in these animals, leptin administration at the same dose that is effective in ob/ob mice is ineffective in reducing food intake or body weight. Perhaps leptin resistance needs to develop in Israeli Sand Rats to allow the development of obesity and, in fact, leptin resistance may be the "thrifty gene" that predisposes individuals to the development of obesity and subsequent metabolic abnormalities. However, there remain many unanswered questions about the physiological actions of leptin. The widespread tissue location of receptors and the actions of leptin independent of food intake highlight the need for further research aimed at determining the major physiological action of this newly discovered and exciting hormone. PMID- 9329742 TI - Diabetes and hypertension in rodent models. AB - As shown by ourselves and others, animals models closely resembling human complex diseases like IDDM in BB/OK and hypertension in SHR/Mol rats can be used to dissect a complex disease into discrete genetic factors as has been done for hypertension in (BB/OK x SHR/Mol) cross hybrids. Discrete genetic factors, so called QTLs, were detected on chromosomes 1, 10, 18, 20, and X. To gain additional information about the physiologic effect of the mapped blood pressure QTLs, genetically defined regions of the SHR rat were transferred onto the genetic background of diabetes-prone BB/OK rats. Four new congenic BB.SHR rats named BB.Sa, BB.Bp2, BB.1K, and BB.Xs were generated and characterized telemetrically for blood pressure, heart rate, and motor activity. The data demonstrate clearly that each single blood pressure QTL of the SHR rat causes a significant increase of the systolic blood pressure and has a different influence on diastolic blood pressure, heart rate, and motor activity. The effects were modified differently by the diabetic state in BB.Sa, BB.Bp2, and BB.Xs rats carrying all diabetogenic genes of the BB/OK rats. The results demonstrate that these newly established congenic strains are a unique tool to study the physiological control of blood pressure by a single blood pressure QTL on the one hand and their interaction with hyperglycemia on the other. It is well within the bounds of possibility that diabetic congenics reflect the diabetic hypertension seen in diabetic patients. Because of the synteny conservation in gene order between different mammals, genes of the appropriate human region could therefore be candidate genes for hypertension in diabetics. Furthermore, these congenic strains can also be used to study interactions between a blood pressure QTL and various selected environmental conditions. In this way, one could learn which QTL can be influenced by environmental factors and to what extent. Another point is the study of gene interactions. Because congenics are genetically identical except for the defined transferred region, congenics can be crossed to investigate the interaction between two or three blood pressure QTLs selected by the investigator and not by nature. These QTL combinations can be studied in the nondiabetic as well as diabetic state. Although the advantage of congenic strains has been shown, the transferred chromosomal regions are too large to pinpoint the gene responsible for the phenotypic change. Therefore, regions on each chromosome must be systematically whittled down, which can be done by crossing the congenics with BB/OK rats and intercrossing their progeny to generate recombinants. These can then be used for the creation of new congenic lines carrying a much smaller region of the SHR/Mol rat. This has been started for the region on chromosome 1 spanning a 16-cM region from the Sa to the Igf2 gene. BB.Sa rats were therefore backcrossed onto BB/OK rats and the resulting progeny were intercrossed. The aim will be to create at least three new congenic BB.Sa rat strains homozygous for the SHR alleles of Sa, Lsn, or Igf2 genes. However, new problems will emerge with these new congenics. To genetically define small regions requires more dense polymorphic markers than are currently available. Dense polymorphic markers will also be necessary to split the other regions on chromosomes 10, 18, 20, and X. We expect that in the near future it will be possible using this approach to define small regions of < 0.5 cM. The recent progress in gene mapping in the rat gives hope that the use of such congenic lines will allow the identification and recovery of the blood pressure genes in the near future. PMID- 9329743 TI - Insulin resistance syndrome in mice deficient in insulin receptor substrate-1. PMID- 9329744 TI - WOK.1W rats. A potential animal model of the insulin resistance syndrome. PMID- 9329745 TI - Lipid transport genes and their relation to the syndrome of insulin resistance. PMID- 9329746 TI - Progress in determining the genes for hypertension, insulin resistance, and dyslipidemia. AB - For the first time, we have techniques available that may enable us to determine cause and effect in common cardiovascular diseases. The observations presented herein require cautious interpretation until replicated. There are currently several large programs under way that seek to establish substantial family resources for investigation of the genetic basis of hypertension. When interpreting the results of genome screens, it will be necessary to consider that we may link genetic loci for coexistent features such as dyslipidemia and insulin resistance to hypertension. Therefore, careful phenotypic data will be necessary to dissect out the causes of hypertension. PMID- 9329748 TI - Is a mutation of the beta 3-adrenergic receptor gene related to non-insulin dependent diabetes mellitus and juvenile hypertension in the Czech population? PMID- 9329747 TI - Hyperinsulinemia and sympathoadrenal system activity in the rat. PMID- 9329749 TI - Glucose transport and insulin signaling in rat muscle and adipose tissue. Effect of lipid availability. PMID- 9329750 TI - Cafestol (a coffee lipid) decreases uptake of low-density lipoprotein (LDL) in human skin fibroblasts and liver cells. PMID- 9329751 TI - High fructose feeding enhances erythrocyte carbonic anhydrase 1 mRNA levels in rat. PMID- 9329752 TI - Pharmacological treatment and mechanisms of insulin resistance. Impact on vascular smooth muscle cells, blood pressure, and lipids. PMID- 9329754 TI - Impact of dietary fatty acid composition on insulin action at the nucleus. PMID- 9329753 TI - Fatty acid regulation of gene expression. Its role in fuel partitioning and insulin resistance. AB - Dietary polyenoic (n-6) and (n-3) fatty acids uniquely regulate fatty acid biosynthesis and fatty acid oxidation. They exercise this effect by modulating the expression of genes coding for key metabolic enzymes and, in doing this, PUFA govern the intracellular as well as the interorgan metabolism of glucose and fatty acids. During the past 20 years, we have gradually elucidated the cellular and molecular mechanism by which dietary PUFA regulate lipid metabolism. Central to this mechanism has been our ability to determine that dietary PUFA regulate the transcription of genes. We have only begun to elucidate the nuclear mechanisms by which PUFA govern gene expression, but one point is clear and that is that it is unlikely that one mechanism will explain the variety of genes governed by PUFA. The difficulty in providing a unifying hypothesis at this time stems from (a) the many metabolic routes taken by PUFA upon entering a cell and (b) the lack of identity of a specific PUFA-regulated trans-acting factor. Nevertheless, our studies have revealed that PUFA are not only utilized as fuel and structural components of cells, but also serve as important mediators of gene expression, and that in this way they influence the metabolic directions of fuels and they modulate the development of nutritionally related pathophysiologies such as diabetes. PMID- 9329755 TI - Molecular and cellular determinants of triglyceride availability. PMID- 9329756 TI - Dietary fatty acids, insulin resistance, and diabetes. PMID- 9329757 TI - Lipid abnormalities in muscle of insulin-resistant rodents. The malonyl CoA hypothesis. PMID- 9329758 TI - Pharmacological strategies for reduction of lipid availability. PMID- 9329760 TI - Relationships between fatty acid composition and insulin-induced oxidizability of low-density lipoproteins in healthy men. PMID- 9329759 TI - Effect of oral antidiabetics and insulin on lipids and coronary heart disease in non-insulin-dependent diabetes mellitus. PMID- 9329761 TI - Small dense low-density lipoprotein (LDL) in non-insulin-dependent diabetes mellitus (NIDDM). Impact of hypertriglyceridemia. PMID- 9329762 TI - Does dietary fat influence insulin action? AB - What is clear from the research thus far is that dietary fat intake does influence insulin action. However, whether the effect is good, bad, or indifferent is strongly related to the fatty acid profile of that dietary fat. The evidence has taken many forms, including in vitro evidence of differences in insulin binding and glucose transport in cells grown with different types of fat in the incubation medium, in vivo results in animals fed different fats, relationships demonstrated between the membrane structural lipid fatty acid profile and insulin resistance in humans, and finally epidemiological evidence linking particularly high saturated fat intake with hyperinsulinemia and increased risk of diabetes. This contrasts with the lack of relationship, or even possible protective effect, of polyunsaturated fats. In particular, habitual increased n-3 polyunsaturated dietary fat intake (as fish fats) would appear to be protective against the development of glucose intolerance. It is reassuring that the patterns of dietary fatty acids that appear beneficial for insulin action and energy balance are also the patterns that would seem appropriate in the fight against thrombosis and cardiovascular disease. Mechanisms, though, still need to be defined. However, there are strong indicators that defining the ways in which changes in the fatty acid profile of membrane structural lipids are achieved, and in turn influence relevant transport events, plus understanding the processes that control accumulation and availability of storage lipid in muscle may be fruitful avenues for future research. One of the problems of moving the knowledge gained from research at the cellular level through to the individual and on to populations is the need for more accommodating research designs. In vitro studies may provide in-depth insights into intricate mechanisms, but they do not give the "big picture" for practical recommendations. On the other hand, correlational studies tend to be fairly blunt instruments, requiring large numbers that are very often not feasible if a greater depth of understanding of the biological processes is to be incorporated. There may be benefit in turning to the clinical case study as a framework for a more comprehensive analysis of the links between dietary fats and insulin action. The real challenge is to keep the depth of analysis rigorous enough to be able to explain and accommodate individual variation (i.e., the diversity of both environmental and genetic backgrounds) while at the same time satisfying the cultural need to provide appropriate overall dietary guidelines. Finally, David Kritchevsky brought to our attention a delightful quote from Mark Twain: "There is something fascinating about science. One gets such a wholesale return of conjecture for such a trifling investment of fact." In the field of dietary fats and the Metabolic Syndrome, this quotation is, unfortunately, apt. Much more research is necessary to define how dietary fats really work to affect insulin action. Well designed, long-term studies in "free range" humans must be undertaken if dietary guidelines for the Metabolic Syndrome are to be based on anything more than a "trifling" amount of "fact." PMID- 9329763 TI - Monounsaturated and marine omega-3 fatty acids in NIDDM patients. PMID- 9329764 TI - Omega-3 and omega-6 fatty acids in the insulin resistance syndrome. Lipid and lipoprotein metabolism and atherosclerosis. AB - Dietary fatty acids appear to be of significant importance for several of the most-common diseases in modern societies. To obtain more knowledge about the health consequences of dietary fatty acids, we depend upon a better understanding of the mechanisms of action of these fatty acids in vivo. With regard to the IRS, omega-3 PUFA may exert beneficial effects upon many of the associated pathophysiological metabolic changes. Omega-3 PUFA reduce fasting and postprandial TG, may improve insulin sensitivity (as shown in animal experiments), decrease platelet and leukocyte reactivity, alter immunological functions, and may slightly decrease blood pressure. Omega-3 PUFA may also beneficially influence vessel wall characteristics and blood rheology. Furthermore, both types of PUFA (omega-3 and omega-6) have been shown to inhibit cardiac arrhythmias in animals. The role of omega-3 PUFA in blood clotting and fibrinolysis still remains controversial, whereas omega-6 fatty acids may lead to increased oxidation of lipoproteins. Regardless of the effects on LDL oxidizability, both types of PUFA have shown beneficial effects on the development of atherosclerosis. As yet, little is known about the effect of specific omega-6 fatty acids with respect to the IRS. Potential adverse effects of dietary PUFA must not be neglected, but should be viewed in light of the beneficial effects of these agents. PMID- 9329765 TI - Omega-6/omega-3 fatty acid ratio and trans fatty acids in non-insulin-dependent diabetes mellitus. PMID- 9329766 TI - Dietary fats and hypertension. Focus on fish oil. PMID- 9329767 TI - Postprandial triglyceride high response and the metabolic syndrome. PMID- 9329768 TI - Evaluation of an omega-3 fatty acid supplement in diabetics with microalbuminuria. PMID- 9329769 TI - Paleodiet and its relation to atherosclerosis. PMID- 9329770 TI - Passive overconsumption. Fat intake and short-term energy balance. PMID- 9329771 TI - Dietary fats and thermogenesis. PMID- 9329772 TI - Fat metabolism in the predisposition to obesity. PMID- 9329773 TI - Fat and energy balance. AB - In summary, an imbalance between energy intake and energy expenditure can explain approximately 80% of the variance in body weight gain in this dietary model of obesity. Several metabolic variables appear to contribute to differences in energy balance. A high RQ and an inappropriate suppression of glucose production by insulin appear to be linked to the increase in energy intake that occurs when obesity-prone rats are provided with the high-fat diet. In addition, early tissue enzymatic differences in obesity-prone versus obesity-resistant rats may contribute to differences in energy expenditure and/or to differences in nutrient partitioning. In this dietary model, susceptibility to dietary obesity involves a metabolic environment that includes a high RQ and a reduced ability of insulin to suppress glucose appearance (FIG. 9). However, this environment does not lead to obesity nor to a measurable difference in body weight gain when the susceptible rats are eating a low-fat diet. The high-fat diet is a necessary catalyst for the observed variability in body weight gain and the development of obesity. As a catalyst, the high-fat diet results in an imbalance between energy intake and energy expenditure in some, but not all, rats. This imbalance interacts with the permissive metabolic environment (tissue enzymatic profile favoring carbohydrate utilization and lipid storage) to produce obesity on the high-fat diet. Later, in the HFD feeding period, the rate of weight gain is not significantly different between OP and OR rats, although net fat accumulation remains greater in the former group. It is interesting that this later period is characterized by a reduction in the difference in both RQ and energy intake between OP and OR rats. Thus, during the later stages of HFD feeding, the discrepancy in both energy balance and nutrient balance between OP and OR rats is reduced. This dietary model of obesity is relevant to human obesity. While the prevalence of obesity is high, the majority of people are not obese. The high prevalence of obesity may be due to environmental catalysts that interact with inherent behavioral and metabolic characteristics that favor nutrient retention. Resistance to obesity can be achieved by avoiding these environmental catalysts, by having inherent characteristics that prevent nutrient retention, or both. Our work suggests that the complete understanding of obesity will require not only the identification and functional significance of the genes that determine the inherent capacity of the behavioral and metabolic systems, but also the role of environmental catalysts in determining where and how these systems operate. PMID- 9329774 TI - Recent advances in the pharmacological control of energy balance and body weight. PMID- 9329775 TI - Fat substitutes and energy balance. PMID- 9329776 TI - Nonesterified fatty acid regulation of lipid and glucose oxidation in the obese. PMID- 9329777 TI - Nuclear all-trans retinoic acid receptors in liver of rats with diet-induced insulin resistance. AB - Retinoic acid receptor alpha (RAR alpha) expression and RAR binding characteristics were investigated in the liver of rats with high-sucrose (HS) diet-induced insulin resistance. Animals were fed a basal (B) or HS (63 cal%) diet with or without fish oil (FO) (30% w/w of total fatty acids) for two weeks. A significant augmentation (p < 0.01) in the RAR alpha mRNA accumulation in rats fed HS diet when compared to rats fed B diet was demonstrated. In comparison with rats fed B + FO diet, a significant increase (p < 0.005) in the RAR alpha expression was found in rats fed HS + FO diet. In [3H]-retinoic acid (RA) binding studies, Scatchard plots confirmed a significant increase (p < 0.05) in the RAR maximal binding capacity (Bmax) only in rats fed HS + FO diet when compared to rats fed B diet. No significant changes in the association constant (Ka) were found among the groups when compared to rats fed B diet. In contrast to RAR alpha, a significant decrease (p < 0.005) in nuclear thyroid hormone receptor alpha 1 (TR alpha 1) expression was found in rats fed HS diet when compared to rats fed B diet. A significant decrease (p < 0.05) in the TR alpha 1 expression was also detected in rats fed HS + FO diet in comparison with rats fed B + FO diet. In addition, an analogous pattern in the expression of the TR isoform (TR alpha 2) was evaluated as well. In conclusion, the high-sucrose diet-induced insulin resistance might be associated with an increased RAR alpha expression and RAR population, and also with a decreased TR alpha 1 and TR alpha 2 mRNA accumulation. PMID- 9329778 TI - Diet-induced insulin resistance is associated with decreased activity of type I iodothyronine 5'-deiodinase in rat liver. AB - The effect of feeding Wistar rats with high-sucrose (63 wt% of sucrose, HS) or high-fat (30 wt% of fat, HF) diets for two weeks on serum selenium concentration and type I iodothyronine 5'-deiodinase (5'-DI) activity in liver was investigated. No significant differences in serum selenium concentration (as determined by graphite-furnace atomic absorption spectrometry) were found among the groups of rats fed basal, HS, or HF diets. A significant reduction of the 5' DI activity (p < 0.005-0.05) was found in groups of rats fed either HS or HF diet in comparison with rats fed B diet. In conclusion, it is suggested that decreased 5'-DI activity in HS or HF diet-induced insulin resistance is not due to selenium status, but it may involve other dietary-related factors. PMID- 9329779 TI - Insulin and catecholamines act at different stages of rat liver regeneration. AB - Insulin and catecholamines are known to exert effects on hepatocyte growth and metabolism. The binding of insulin, the plasma levels of insulin (INS), and the plasma catecholamine levels of epinephrine (EPI) and norepinephrine (NE) were measured during liver regeneration after partial hepatectomy (PH). A significant decrease (p < 0.05) of INS receptor binding capacity was found at 1, 2, and 3 days after operation. A single insulin injection (2.5 IU/kg body weight) at 24 h after sham operation or partial hepatectomy did not affect these changes of INS binding to hepatocytes. The plasma insulin and glucose levels were similar in both hepatectomized and sham-operated rats. Within 20 min after liver resection or sham operation, plasma NE and EPI concentrations increased rapidly. Then, a significant decrease was observed in plasma catecholamine levels at 1 h after laparotomy and PH. In both groups, laparotomized and partially hepatectomized plasma levels of NE at 4 h reached control values and remained unchanged at the 4 and 24-h periods. After PH, the levels of EPI remained elevated at 4 h in comparison with laparotomy. Adrenal tyrosine hydroxylase mRNA levels were significantly elevated at 4 h in both PH and sham-operated groups. These results suggest that signals that are initiated by catecholamines and transduced through second messengers presumably participate in the trigger mechanism of liver regeneration, while insulin (considered as a secondary mitogen) enhances a stimulus for liver regeneration. PMID- 9329780 TI - Fatty acid composition in fractions of structural and storage lipids in liver and skeletal muscle of hereditary hypertriglyceridemic rats. AB - The fatty acid (FA) compositions of liver and skeletal muscle structural lipids, overall phospholipids and phosphatidylcholine, and triglycerides (TG) were determined in the hereditary hypertriglyceridemic (HTG) rat, a nonobese animal model of the insulin resistance syndrome. Four groups of HTG rats and four groups of control animals were fed equal-energy diets for two weeks: basal (B), high sucrose (HS), or fish oil-supplemented basal (BFO) or high-sucrose (HSFO) diets. In the liver of HTG rats, a decrease of n-6 long-chain polyunsaturated FA (PUFA), especially in 20:4n-6, in comparison with controls was found. Moreover, a concomitant accumulation of 18:2n-6 in structural lipids was observed. These differences were more pronounced in liver than in skeletal muscle. HS feeding raised the proportion of 18:1n-9 and decreased 18:2n-6 in lipid fractions. In both tissues and in both strains, the amounts of long-chain n-3 PUFA, as well as the level of total C20-22 PUFA, went up after fish oil feeding. However, the effects were somewhat less pronounced in the HTG rats. The increase in n-3 PUFA occurred mainly at the expense of reduced levels of 18:2n-6 in structural lipids and of 18:1n-9 in triglycerides. These changes were associated, in companion studies reported in this volume, with improved insulin action in HTG rats. In conclusion, the FA composition in lipid subclasses of HTG rats differs significantly from the controls mainly in liver structural lipids, suggesting the impairment of PUFA desaturation. Dietary change effected a similar modulation of FA profile across both strains, with fish oil increasing the levels of long-chain PUFA toward control values in the NTG rats. The HTG rat thus provides an interesting animal model for the study of impaired fatty acid metabolism. PMID- 9329781 TI - Increased adipose tissue lipolysis in a hypertriglyceridemic rat line. PMID- 9329782 TI - Structural changes in the aorta of the hereditary hypertriglyceridemic rat. AB - Structural changes in the ascending thoracic aorta of hereditary hypertriglyceridemic (hHTG), insulin-resistant, and hypertensive rats were studied using transmission electron microscopy. Normotensive Wistar rats were used as controls. The most-pronounced morphological changes were observed in the tunica intima. Endothelial cells of hHTG rats formed a continual layer around the whole circumference. Subendothelial space was enlarged. Some endothelial cells were delaminated from the subendothelial space by big lipid droplets that were often present in the subendothelial space, and the endothelial cells bulged out towards the lumen. Big electron-lucent lipid droplets were present in the majority of the endothelial cells and occupied the main part of the cytoplasm. Degenerative microvesicular and membranous material was present in the cytoplasm. Increased numbers of vesicles of Golgi apparatus and cisternae of endoplasmic reticulum were found. Similar morphological alterations, but in less-extended form, were observed in smooth muscle cells. The organization and orientation of smooth muscle cells were essentially intact. In muscle cells, lipid droplets were localized in close relation to Golgi complex and in dilated cisternae of the sarcoplasmic reticulum. Lipid droplets, degenerative material, myelin figures, myelinoid membranes, and vesicular components were also sporadically found in the intercellular space among muscle cells. This pilot morphological investigation provides further arguments for a thorough and more-focused electron microscopy study of conductance arteries of the hHTG rats. PMID- 9329783 TI - Increased lipoprotein oxidability and aortic lipid peroxidation in an experimental model of insulin resistance syndrome. PMID- 9329784 TI - Phenotype and genotype comparison of hereditary hypertriglyceridemic (hHTG) and brown-Norway (BN) rats. Identification of quantitative trait loci (QTLs) for the insulin resistance syndrome. PMID- 9329785 TI - Insulin resistance in adipocytes of SHR rats. PMID- 9329786 TI - Disproportionate increase of fatty acid binding proteins in the livers of obese diabetic Psammomys obesus. PMID- 9329787 TI - Partial characterization of insulin resistance in adipose tissue of monosodium glutamate-induced obese rats. PMID- 9329788 TI - Biguanide effects on insulin signaling. PMID- 9329789 TI - Effect of the high-fat diet on the calcium channels in rat myocardium. PMID- 9329790 TI - The effect of fasting and vitamin E on insulin action in obese type 2 diabetes mellitus. AB - The influence of either short-term fasting or vitamin E administration on insulin action was studied in two groups of obese Type 2 diabetic patients. Twelve patients underwent 7 days of fasting (group A), whereas 600 mg of vitamin E was administered daily during 3 months in 9 diabetic patients (group B). Insulin action was examined by using hyperinsulinemic isoglycemic clamps (insulin infusion rate, 1.0 mU/kg/min) and insulin receptors on erythrocytes before and after respective regimens. An increase of glucose disposal rate (29.5 +/- 8.9 vs. 24.0 +/- 7.5 mumol/kg/min, p < 0.01) and an increase of metabolic clearance rate of glucose (4.0 +/- 2.5 vs. 2.3 +/- 0.9 mL/kg/min, p < 0.01) were observed in group A after fasting. On the contrary, decreases of glucose disposal rate (21.3 +/- 8.5 vs. 26.6 +/- 9.8 mumol/kg/min, p < 0.02), metabolic clearance rate of glucose (2.9 +/- 0.8 vs. 3.7 +/- 1.7 mL/kg/min, p < 0.05), and insulin receptor number (p < 0.01) were found after vitamin E administration as compared with pretreated values. A worsening of diabetes control as observed by an increase of HbA1C (p < 0.01) was present in the latter group. In summary, we found an improvement of insulin action after short-term fasting in contrast with the worsening of metabolic parameters after vitamin E administration in obese Type 2 diabetic patients. PMID- 9329791 TI - The effect of hyperlipidemia on serum fatty acid composition in type 2 diabetics. AB - Fatty acid (FA) profiles of total serum lipids were determined by capillary gas chromatography in Type 2 diabetic patients (NIDDM) with diverse types of hyperlipidemia. In patients with hypertriglyceridemia (DM-HTG) and combined hypertriglyceridemia and hypercholesterolemia (DM-HLP), a significantly different total FA composition was found compared with healthy controls or diabetics with normal serum lipids. In particular, the proportions of saturated and monounsaturated FA were increased and the proportions of n-6 polyunsaturated FA were decreased. In DM-HLP patients, PUFA n-6 metabolites and C20-C22 PUFA were also decreased. Thus, hyperlipidemia shifts significantly the serum FA composition in NIDDM patients into an atherogenic profile. More study is needed, however, to understand if serum FA changes may contribute to the increased atherogenesis commonly found in these patients. PMID- 9329792 TI - High lipid levels in Slovak rural population. Consequence of thyroid dysfunction or nutritional status? PMID- 9329793 TI - Activity of antioxidant enzymes during hyperglycemia and hypoglycemia in healthy subjects. PMID- 9329819 TI - Female reproductive science in historical perspective. PMID- 9329821 TI - Differentiation-dependent and cell-specific regulation of the hIGFBP-1 gene in human endometrium. AB - We analyzed IGFBP-1 gene promoter activity by transient transfection during the progressive decidualization of human endometrial stromal cells. A time study over a 13-day culture period showed that the promoter activity increased exponentially to > 10(4) fold in cells treated with MPA and RLX correlating with the secretion rate and steady-state mRNA levels of the endogenous gene. Deletion analysis showed that two regions in the IGFBP-1 gene promoter are responsible for the activation of the IGFBP-1 gene. The basal promoter region between -1 and -300 bp contains multiple sections of functional elements homologous either to CRE, PRE, or CCAAT. The major difference of IGFBP-1 gene activation in endometrium and the hepatic system lies in the distal promoter region, between -2.6 and -3.4 kb, which mediates 95% of the total promoter activity derived from -3.3 kb to +68 bp. Functional and binding analysis in the distal promoter region showed that multiple Sp1 elements interacting with a novel Sp3 transcription factor activates the hIGFBP-1 gene promoter. PMID- 9329820 TI - Differential expression of the A and B isoforms of progesterone receptor in human endometrial cancer cells. Only progesterone receptor B is induced by estrogen and associated with strong transcriptional activation. PMID- 9329822 TI - Nestorone progestin. The ideal progestin for use in controlled release delivery systems. PMID- 9329823 TI - RU 486 (mifepristone). A short overview of its mechanisms of action and clinical uses at the end of 1996. AB - RU 486 (mifepristone) has proved to be a remarkably active antiprogesterone and antiglucocorticosteroid agent in humans. The mechanism of action of RU 486 involves the intracellular receptors of the antagonized hormones progesterone and glucocorticosteroids. At the molecular level, the most important features are high binding affinity to the receptor, interaction of the phenyl-aminodimethyl group in the 11 beta-position with a specific region of the receptor binding pocket, and RU 486-induced transconformation in the ligand binding domain. These properties have consequences at different steps of the receptor function as compared with agonists. However, this cannot be limited to the RU 486-receptor interaction; for instance, a switch from an antagonistic property to an agonist activity is possible, depending on the intervention of other signaling pathways. Derivatives with only one of the two antagonistic properties (antiprogestin, antiglucocorticosteroid) would be desirable in spite of similarities between the steroid structures, receptors involved, and responsive machineries in target cells. Clinically, the RU 486 plus prostaglandin method is ready to be used on a large scale, and is close to being as practical and safe as any medical method of abortion may be. The early use of RU 486, as a contragestive--that is, for use by a woman as soon as she fears a pregnancy she does not want--will help to defuse the abortion issue. Research should now be conducted to define an efficient and convenient contraceptive method with RU 486 or other antiprogestins. The usefulness of RU 486 for obstetrical indications, including facilitation of difficult delivery, has to be assessed rapidly. Gynecological trials, particularly in leiomyomata, should also be systematically continued. The very preliminary results obtained with tumors, including breast cancers, do indicate that further studies are necessary. PMID- 9329825 TI - Endometrial corticotropin-releasing hormone. Its potential autocrine and paracrine actions. AB - Corticotropin-releasing hormone (CRH) is expressed at several peripheral tissues including normal epithelial cells of human and rodent uterus. However, its biological role is unknown in both species. To clarify this role we studied the regulation of CRH promoter in endometrial cells. We performed homologous transfection experiments in Ishikawa cells, a human endometrial cell line, using a 0.9-kb fragment of the 5'-flanking region of human CRH gene coupled to luciferase. We found that the activity of the 5'-flanking region of the CRH gene is stimulated by cAMP and EGF and inhibited in a receptor-mediated, dose dependent fashion by estradiol and dexamethasone. The antiglucocorticoid RU 486 acted as a glucocorticoid agonist suppressing CRH gene activation, whereas progesterone was devoid of any activity. Prostaglandin E2 and interleukins-1 and 6 stimulated CRH activation, and the prostanoid inhibitor indomethacin suppressed it, most probably by inhibiting endogenous prostaglandins. These findings suggest that endometrial CRH gene expression may be under the negative control of estrogens and glucocorticoids and under the positive control of PGE2, IL-1, and IL-6. Considering the involvement of CRH in proinflammatory phenomena, we postulate that endometrial CRH, in association with uterine prostanoids and cytokines, may participate in intrauterine inflammatory processes of early pregnancy, such as decidualization and blastocyst implantation. PMID- 9329824 TI - The endometrial approach in contraception. PMID- 9329826 TI - In vitro bioassays for hormonal activities. PMID- 9329827 TI - Antiprogestin action on the endometrium. PMID- 9329829 TI - Integrins and the endometrium: new markers of uterine receptivity. PMID- 9329828 TI - A review of the endometrial changes in Norplant users. PMID- 9329831 TI - Implantation: from basics to the clinic. PMID- 9329830 TI - Recombinant virus-mediated gene transfer in trophoblast cells. PMID- 9329832 TI - Interleukin-1 alpha opposes progesterone-mediated suppression of MMP-7. A possible role of this cytokine during human implantation. PMID- 9329833 TI - Multifaceted roles for IGFBP-1 in human endometrium during implantation and pregnancy. AB - IGFBP-1 is a major protein product of nonpregnant endometrium during the mid-late secretory phase and occurs in abundance in decidua. Its roles as an IGF-binding protein and as a trophoblast integrin ligand suggest that it may have multiple roles in endometrial development and in interactions between the decidua and the invading trophoblast. IGFBP-1 in vaginal/cervical secretions has already had clinical application as a predictor of premature rupture of fetal membranes. The future awaits elucidation of the potential utility of IGFBP-1 in serum and in decidua in predicting fetal growth restriction and preeclampsia and perhaps implantation failure. PMID- 9329834 TI - Use of recombinant leukemia inhibitory factor in embryo implantation. PMID- 9329835 TI - Update in preimplantation genetic diagnosis. Age, genetics, and infertility. AB - PGD has been successfully used for several years. Over 40 babies have been born worldwide by use of these techniques. Unfortunately, a number of misdiagnoses have been made, a distressing consequence of a new frontier. Significant advances have been made to improve the efficiency and accuracy of PCR and FISH. The widespread use of this technology awaits further documentation of safety and accuracy. Other issues must also be addressed. First, the cost-effectiveness of the techniques relative to the traditional alternatives must be evaluated. A number of ethical issues regarding embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for nongenetic disorders such as gender selection. Finally, the experimental nature of these procedures must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved IVF success rates may make PGD a more widely used clinical tool. The future awaits these developments. PMID- 9329836 TI - Infertility and early pregnancy loss is largely due to oocyte aging, not uterine senescence, as demonstrated by oocyte donation. PMID- 9329837 TI - Physiological and clinical implications of decidualization-associated protease activity. PMID- 9329838 TI - Hypoxia stimulates ecNOS mRNA expression by differentiated human trophoblasts. AB - Cytotrophoblasts isolated from normal human placenta cultured under normoxic conditions (20% O2, pO2 = 130 mmHg) for 48-72 h differentiate to a form which expresses high levels of hCG and which morphologically resembles syncytiotrophoblast. We had previously shown that hypoxia (0-1% O2, pO2 = 12-14 mmHg) blocks this differentiation process, although trophoblasts exposed to hypoxia for up to 96 h were completely viable. In this article we showed that trophoblast responds to hypoxia by expressing the hypoxia-sensitive DNA binding protein HIF-1. We also showed that in trophoblast cultured under normoxic conditions, expression of endothelial cell nitric oxide synthase (ecNOS) mRNA increases with time, reaching a maximum in 48-72 h. However, in trophoblast maintained under hypoxic conditions for 48 h (after an initial 24 h in normoxia), expression of ecNOS mRNA is greatly reduced. These observations are consistent with the expression of ecNOS by syncytiotrophoblast but not by cytotrophoblast. In contrast, exposure of differentiated trophoblasts to hypoxia for 24 h (after 48-72 h in normoxia) significantly stimulates expression of ecNoS mRNA over that of cells maintained continuously in normoxia. These results suggest that in differentiated trophoblast hypoxia can stimulate ecNOS expression. PMID- 9329839 TI - Decidual cell regulation of hemostasis during implantation and menstruation. AB - Progesterone stimulation of the estradiol (E2)-primed human endometrium initiates DZ of the stromal cells around the spiral arterioles. Under continued steroid stimulation, DZ spreads wave-like to establish the decidual cell as a major cell type of the luteal phase and pregnant endometrium. Because of their widespread distribution throughout the endometrium and concentration at perivascular sites, decidual cells are spatially and temporally positioned to mediate the opposing requirements of maintaining hemostasis during endovascular trophoblast invasion, yet promoting menstrual hemorrhage in the absence of implantation. The experimental results summarized in this review indicate that the paradoxical properties manifested by endometrial stromal/decidual cells are controlled by several proteins with either hemostatic or ECM-degrading or vasoactive activity, and that their expression is altered in response to changes in levels of circulating ovarian steroids during the menstrual cycle. These conclusions are drawn primarily from studies with a well-characterized in vitro model of DZ using monolayers of stromal cells derived from specimens of predecidualized endometrium. Thus, progestins modify the expression of several DZ-related markers in the cultured stromal cells, and E2 enhances these effects despite the lack of response to E2 alone. These responses are consistent with the differential actions displayed by E2 and progesterone in vivo, by which E2 primes the endometrium for the decidualizing effects of progesterone by elevating progesterone receptor levels. Accordingly, during steroid-induced in vitro DZ, a marked increase in the expression of stromal cell TF and PAI-1 and reciprocal inhibition of tPA activity suggest mechanisms to account for the absence of hemorrhage during invasion of the endometrial vasculature by implanting trophoblasts. In contrast to steroid-induced DZ, the events of menstruation are initiated in response to a decline in circulating levels of ovarian steroids. Accordingly, subjecting in vitro decidualized stromal cells to steroid withdrawal results in pronounced reversal in the expression of all of the end points listed above. Consequently, the local hemostatic environment is transformed into a hemorrhage-promoting milieu. Taken together with vascular injury resulting from ischemia induced by spiral artery vasoconstriction, the net effect is attainment of two prerequisites for menstrual hemorrhage, vascular injury and inadequate hemostasis. PMID- 9329840 TI - Angiogenesis and macrophage activation in endometriosis. AB - Recent studies suggest that the symptoms associated with endometriosis are the result of local peritoneal inflammation. Increased concentrations of activated pelvic macrophages and lymphocytes and the elevated levels of specific cytokines and growth factors reviewed above support this hypothesis. The precise roles of these soluble factors are currently unknown, but we propose that a complex network of endometrial cytokines are normally regulated by hormones produced during the ovulatory cycle. Ectopic endometrial implants also are subject to these same endocrine cues. The secretion of these proinflammatory proteins by endometriosis lesions into the peritoneal microenvironment appears to cause a recruitment of capillaries and activated inflammatory cells to the implant. Future therapeutic strategies directed to ameliorate the inflammatory reaction associated with endometriosis should not ignore the likely physiological actions of many of the same bioactive molecules in normal eutopic endometrial function. PMID- 9329841 TI - Evaluation of abnormal vaginal bleeding in perimenopausal women with endovaginal ultrasound and saline infusion sonohysterography. AB - Saline infusion sonohysterography enhances endovaginal ultrasound examination of the uterine cavity in perimenopausal patients with abnormal uterine bleeding. It is easily and rapidly performed at minimal cost, is extremely well tolerated by patients, and is virtually devoid of complications. Its use can prevent invasive diagnostic procedures in some patients as well as optimize the preoperative triage process for those patients who will require therapeutic intervention. PMID- 9329842 TI - Contractility in the nonpregnant uterus. PMID- 9329843 TI - Abnormal uterine contractility in nonpregnant women. PMID- 9329844 TI - Placental stress factors and human parturition. PMID- 9329845 TI - Regulation of the uterus and cervix during pregnancy and labor. Role of progesterone and nitric oxide. PMID- 9329846 TI - Prostaglandins and parturition. PMID- 9329847 TI - Role of Fas ligand in conferring immune privilege to non-lymphoid cells. PMID- 9329848 TI - The endocrinology of human parturition. PMID- 9329849 TI - Vaginal drug delivery: the first uterine pass effect. PMID- 9329850 TI - The first uterine pass effect. AB - The endometrial effects of vaginal progesterone have been found to be unexpectedly reliable. This has led us to suspect that a local direct vagina-to uterus transport or first uterine pass effect was the basis of the uterine targeting of vaginal progesterone. After vaginal administration of progesterone, uterine tissue concentration has been found to exceed by more than 10-fold the levels achieved by systemic administration, despite plasma levels in the latter case that were more than seven times higher. Similar differences in systemic-to uterine tissue level ratios have been observed between oral and vaginal administration of danazol. Originally seen as a pharmacological advantage permitting the uterine targeting of vaginally administered substances, it is possible that the first uterine pass effect plays a physiological role in the control of uterine contractile activity through the prostaglandins contained in the semen. PMID- 9329851 TI - Ectopic pregnancy. PMID- 9329852 TI - Laparoscopic surgery and assisted reproductive techniques. Combined strategies of therapy in infertile women. PMID- 9329853 TI - Laparoscopic myomectomy. Operative technique and results. PMID- 9329854 TI - Surgery for endometriosis of the bowel, bladder, ureter, and diaphragm. PMID- 9329855 TI - Total laparoscopic hysterectomy. Indications, results, and complications. PMID- 9329856 TI - Transdermal progestins in hormone replacement therapy. PMID- 9329857 TI - Sex hormone-binding globulin in estrogen-dependent cancer and estrogen replacement therapy. PMID- 9329858 TI - Effects of using vasectomized bulls in artificial insemination practice on the reproductive efficiency of Italian buffalo cows. AB - The effects of the presence or absence of vasectomized male buffaloes on the reproductive efficiency of buffalo cows (n = 396) undergoing artificial insemination (AI) was studied on six farms owned and operated by a single consortium. Lactating animals were separated into two groups of various sizes on each farm and kept under semi-range conditions. Vasectomized bulls were present in one group at a bull/empty-cow ratio of 1:30. No bulls were present in the other group. Reproductive efficiency between the two groups over a period of 3.5 months was compared and evaluated on the basis of: 1) the number of spontaneous overt estruses associated with either feeble or intense signs of estrous behaviour; 2) the number of functional estrous cycles, i.e. estrous cycles with luteal phases defined as normal, based on specified progesterone concentrations in milk or blood plasma 8-10 days after estrus; 3) the number of consecutive functional estrous cycles in cases of induced estrus; and 4) pregnancy rate. Groups with bulls present demonstrated a significantly higher reproductive efficiency than groups without them. There was a higher incidence of spontaneous estrus (92 versus 69%; P < 0.01); spontaneous estrus of high intensity (62.2 versus 31.1%; P < 0.01); and higher incidence of functional estrous cycles following both spontaneous (65.8 versus 57.1%) and induced (77.0 versus 59.5%; P < 0.05) estrus. Exposure to vasectomised bulls also increased the incidence of consecutive functional estrous cycles (90.5 versus 68.1%; P < 0.01), and the pregnancy rate in cows inseminated at spontaneous (42.5 versus 18.9%; P < 0.01) or induced (51.1 versus 33.3%; P < 0.05) estrus. Overall pregnancy rate did not differ significantly between cows inseminated at induced or spontaneous estrus, although in the absence of bulls, pregnancy rate per AI was higher in cows inseminated at induced than at spontaneous estrus (33.3 versus 18.9%). PMID- 9329859 TI - Effects of estradiol-17 beta and hCG supplementation on superovulatory responses and embryo quality in swamp buffalo (Bubalus bubalis) implanted with norgestomet. AB - Twenty-four cycling swamp buffaloes with normal reproductive histories and 2-3 months postpartum were used to investigate the effect of addition of estradiol-17 beta and human chorionic gonadotrophin (hCG) to the superovulation regime on the level of ovarian stimulation and embryo production. The estrous cycles of buffaloes were synchronized by prostaglandin injection and then divided into two groups for superovulatory treatment. Those in Group 1 (n = 12) received a implant containing 3 mg norgestomet (Syncro-Mate-B) for 9 days (insertion day is Day 0), with 4000 IU of equine chorionic gonadotrophin (eCG) and 500 micrograms cloprostenol i.m. given at Day 7. Group 2 (n = 12) received the same regime as Group 1, together with 7.5 mg estradiol-17 beta given in three intramuscular injections on Days 3, 5 and 7 in decreasing doses (4.0, 2.5 and 1.0 mg, respectively) and 5000 I.U hCG i.v. coincidentally with the first insemination. Estrus was monitored visually and by placing treated animals with bulls. Each animal was inseminated twice with frozen sperm after standing estrus. The numbers of corpora lutea (CL) and follicles greater than 8 mm in diameter were recorded via palpation per rectum at 6 days after implant removal. Two days later 11 animals from Group 2 and two from Group 1 were slaughtered for direct observation of ovarian responses and for embryo collection. PMID- 9329860 TI - Oocyte retrieval and histological studies of follicular population in buffalo ovaries. AB - Ovaries (N = 250) from slaughtered buffaloes were collected to study follicular population and compare methods of oocyte retrieval. The number and size of surface follicles were recorded and grouped into different categories. Different sized follicles in relation to oocyte diameter were studied histologically. Yield of oocytes per ovary were less (P < 0.05) from ovaries bearing a corpus luteum (CL). Techniques used for oocyte recovery included slicing, follicle puncture and aspiration. The oocyte recovery rate was greatest (P < 0.05) using slicing. The average number of visible surface follicles was 5.20 +/- 0.97 with mean numbers of 2.5, 1.2, 0.82 and 0.62 per ovary for follicles sized 4, 8, 12 and 12mm respectively. Histological studies revealed large numbers of primordial follicles in prepubertal and atretic follicles in senile buffaloes. They also established a biphasic relationship of growth between oocyte diameter and follicular size. PMID- 9329861 TI - Evidence for a role of the ovarian surface epithelium in the ovulatory mechanism of the sheep: secretion of urokinase-type plasminogen activator. AB - The extent of dissolution of tissues within the apical wall of the preovulatory ovine follicle (formative site of rupture) is greater than that of the counterpart basal hemisphere. It has been hypothesized that proteolytic enzymes released from contiguous ovarian surface epithelial cells contribute to apical follicular weakening and ovulation. Ovulation occurs from the dominant ovarian follicle of proestrous ewes at approximately 24 h after administration of luteinizing hormone-releasing hormone (LHRH). Follicular rupture was inhibited in sheep in which the ovarian surface epithelium was surgically removed at 8 (but not at 16) h following LHRH. Plasminogen activator bioactivity was greater within the follicular apex compared to basal wall at 12 h; this difference was negated by prior removal of epithelium at 8 h after LHRH. A low M(r) plasminogen activator of the urokinase-type (uPA) was secreted by epithelial cells recovered from the surface of preovulatory follicles (Western blot analysis). Ovarian epithelium, not associated with a preovulatory follicle, produced very little uPA. Finally, ovulation was suppressed by intrafollicular injection (8 h post LHRH) of uPA antibodies. It is suggested that secretion of uPA by ovarian surface epithelium and consequent plasmin up-regulation within neighboring tunica albuginea and follicular theca is a contributing factor in the mechanism of ovulation. PMID- 9329862 TI - Factors affecting age at first lambing in Yankasa ewes. AB - Data on 133 primiparous Yankasa ewes covering a period of 8 years (1983-1991) were analysed by least squares method for the effects of season, year, litter size, birth weight, weaning age and weaning weight on age at first lambing (AFL). Results showed the AFL (+/-SE) to be 533.1 +/- 10.3 days. Only the effect of season on AFL was significant (P < 0.01) while year of lambing, birth weight, litter size, weaning age and weaning weight did not affect AFL significantly (P > 0.01). AFL was 476.9 +/- 23.3, 523.9 +/- 19.2, 549.8 +/- 19.7 and 581.9 +/- 19.5 days for late wet, early dry, late dry and early wet season born ewes respectively. AFL was significantly (P < 0.01) lowest for ewes born in the late wet season and highest for ewes born in the early wet season. There was no significant (P > 0.01) difference in AFL between ewes born in the early dry season and ewes born in the late dry season, and between late dry and early wet season born ewes. PMID- 9329863 TI - Metalloproteinase activity during growth, maturation and atresia in the ovarian follicles of the goat. AB - Metalloproteinases are an important group of hydrolytic enzymes which participate in interstitial matrix degradation during tissue remodelling processes and therefore may be required during follicular growth and maturation. The activity of metalloproteinases (collagenases, gelatinase, and Pz-peptidase), was measured during growth, maturation and atresia of goat antral follicles. These follicles (n = 67) were separated by size and also classified into four groups: non-atretic (Group I); early atretic (Stage I) (Group II); moderately atretic (Stage II) (Group IIIa); and, late atretic (Stage III) (Group IIIb). Pz-peptidase was greater in granulosa than in thecal cells, and almost absent in follicular fluid. In non-atretic follicles, activity in granulosa cells increased with increasing follicle size, whereas activity peaked in 3-6 mm follicles in thecal cells. Atresia was associated with declining activity in thecal cells from follicles in the 3-6 mm range and in granulosa cells from the > 6 mm range. Interstitial collagenase activity was significant and similar in granulosa and thecal cell extracts and low in follicular fluid from non-atretic follicles. Activity increased significantly in thecal cells, but decreased significantly in granulosa cells from large (> 6 mm) non-atretic follicles. Atresia was associated with declining activity in both types cells and increasing activity in follicular fluid. Gelatinase activity was some times associated with five regions corresponding to molecular weights of 22.1, 30.7, 39.6, 63.8 and 71.4 kDa, and rarely at 91.3 and 81.2 kDa. Overall activity declined with atresia in thecal cells from follicles in the 3-6 mm range, but not in those > 6 mm. In granulosa cells from follicles 3-6 mm, activity varied widely with stage of atresia, while in cells from follicles > 6 mm, activity was greatly increased in atretic follicles. PMID- 9329864 TI - Effects of testosterone and oestradiol on [3H]-thymidine incorporation by porcine granulosa and theca cells. AB - Experiments were carried out to investigate the effects of varying physiological concentrations (0, 10, 100, and 1000 ng ml-1) of oestradiol or testosterone on [3H]-thymidine incorporation by porcine granulosa and theca cells in vitro. Granulosa cells only were recovered from small (1-3-mm) follicles and both granulosa and theca cells recovered from large (4-8-mm) porcine follicles. Cells were cultured for 72 h in medium containing 10% foetal calf serum, 24 h in serum free medium, and finally 40 h in serum-free medium containing [3H]-thymidine and appropriate steroid treatment. Although DNA per well was significantly higher (P < 0.05) at the end of culture in the theca cells than in the granulosa cells, neither steroid treatment had a significant (P > 0.1) effect on DNA concentration in either cell type. Overall, cells from small follicles incorporated significantly (P < 0.01) more [3H]-thymidine than those from medium follicles. Both oestradiol and testosterone significantly (P < 0.01) inhibited thymidine incorporation by cells from both follicle size categories, with a significant (P < 0.05) hormone X dose interaction. Finally, there was a highly significant (P < 0.001) interaction between the response of cells to different hormone concentrations and the follicle size from which they were recovered. These results indicate that both oestradiol and testosterone may act in an autocrine/paracrine manner to inhibit proliferation and encourage differentiation in follicular cells and thus are likely regulators of the later stage of antral follicle development in the pig. PMID- 9329865 TI - Pregnancy diagnosis in dairy goats using progesterone assay kits and oestrous observation. AB - Two qualitative on-farm milk progesterone test kits were used for early pregnancy diagnosis in goats. One kit was based on latex agglutination (LA) and the other on enzyme-linked immunosorbent assay (ELISA). The accuracy of early pregnancy diagnosis by these two methods was compared with the accuracy of oestrous observation (OeO) and the level of plasma progesterone measured by radioimmunoassay (RIA). Dairy goats (n = 73) from a single herd were used for collection of milk and blood samples 20 days after breeding. Ultrasound examination 50 days after mating found that 49 (67%) goats were pregnant, and 24 (33%) were not pregnant. Using ultrasound as a reference method, the accuracy of early pregnancy diagnosis by RIA and OeO was 92% and 86%, respectively, and for non-pregnancy 100% for both RIA and OeO. The accuracy of early pregnancy diagnosis by ELISA and LA was 82% and 79% and for non-pregnancy, 88% and 100%, respectively. The kappa-statistic for RIA, OeO, ELISA and LA was 0.93, 0.84, 0.77 and 0.73, respectively. It was concluded that LA and ELISA tests can be used for early pregnancy diagnosis in dairy goats. However, in the herd studied, early pregnancy diagnosis by OeO was as good as that achieved with progesterone determination using the kits. PMID- 9329866 TI - Vaginal temperature is not related to the time of ovulation in sows. AB - The relationship between vaginal temperature and ovulation time was studied in sows. The vaginal temperature was measured continuously between Day 4 and Day 10 after Altrenogest-treatment in 10 sows. Oestrus was checked with a vasectomized boar at 8-h intervals, and during oestrus, ovulation time was checked with transrectal ultrasonography at 2-h intervals between 07:00 h and 23:00 h. Two sows ovulated between 23:00 h and 07:00 h, and these sows were taken out of the experiment. In the eight remaining sows, a clear day/night rhythm in vaginal temperature was found: between 03:00 h and 09:00 h, vaginal temperature (LSM +/- sem, corrected for sow) was on average 38.2 +/- 0.01 degrees C; between 15:00 h and 21:00 h, vaginal temperature was on average 38.5 +/- 0.01 degrees C (P < 0.001). Between 4 days before ovulation and 2 days after ovulation, no changes in temperature could be found that were related to ovulation time in any of the sows. Therefore, in sows, changes in vaginal temperature cannot be used as a predictor for ovulation time, and consequently cannot be used to predict the best time for insemination. PMID- 9329867 TI - Epidemiology of advanced lung disease in the United States. AB - ALD affects a large segment of the population in the United States, both old and young, and men and women of all races and ethnicities. Many chronic diseases inevitably advance to a stage that results in significant respiratory impairment and disability. Although the causes of some of the diseases are known and the diseases may be preventable, the overall absolute burden of illness in the population is rising because of an enlarging population and newer therapeutic approaches. This is evident despite the lack of consistent and comparable data estimates for all diseases from national database resources. Where the data exist, it is evident that the cost related to the morbidity and mortality of these illnesses is substantial and consumes a significant proportion of health care expenditures. Both morbidity and mortality estimates, as well as cost estimates, are conservative and are likely underestimates of the true overall impact of ALD on the US economy. PMID- 9329868 TI - Natural history and prognosis of advanced lung disease. AB - Large gaps exist in our knowledge of the natural history of advanced lung disease and of the impact of various therapies upon prognosis and survival. Applying the results of population-based epidemiologic studies or limited clinical trials to a specific patient is hazardous because of marked individual variation in survival, even with the most grim of prognoses. Obtaining such prognostic information is essential, however, in addressing current key issues in advanced lung disease-the efficacy of various therapies, timing lung transplantation, referring to hospice care, providing palliative therapy, and determining medical futility. PMID- 9329869 TI - Measurement of dyspnea and quality of life in advanced lung disease. AB - An important aspect of disease management is the ability to measure and report health outcomes. For advanced lung diseases, both dyspnea and HRQOL are key clinical outcomes of interest to patients and health care providers. There are two major reasons to measure those constructs: to differentiate between individuals (or populations) who have varying degrees of the quality (i.e., more dyspnea or better HRQOL); and to evaluate how much dyspnea or HRQOL has changed as a result of a specific intervention or therapy (e.g., LVRS or transplantation). Dyspnea can be measured by clinical instruments-usually questionnaires-that consider various dimensions or components that affect the individual's breathlessness and by direct ratings during a physical task or exercise test. Although multidimensional instruments provide an "indirect" measure of dyspnea, those scales have been tested extensively and have demonstrated significant improvements with a variety of treatments for patients with advanced lung diseases. Another approach is to instruct the patient to rate dyspnea during an exercise test, then examine the full range of dyspnea responses throughout exercise. Disease-specific and generic instruments are available to measure HRQOL. For clinical trials evaluating a new therapy or procedure, disease specific measures are more appropriate because patients and clinicians find the items more relevant. Furthermore, there is greater potential for demonstrating a significant change with a disease-specific instrument (i.e., it is more responsive). If a clinical outcome of a treatment currently exists, however, then a generic HRQOL instrument may be used to provide complementary information. Such data may expand the impact or scope of the therapy, and previously unrecognized adverse effects may be detected. Another advantage of a generic measure is that comparisons can be made across conditions and populations. Clearly, the selection of the HRQOL instrument depends, in large part, on the objective or question of the clinical trial. An essential consideration for both dyspnea and HRQOL instruments is clinical applicability (i.e., can the questionnaire be used in the daily practice of medicine?). Key factors include the number of items to be completed, the possible need for an interviewer, the time for the patient to complete the instrument, and the ability to computerize scoring and results. Although shorter versions of established "full" instruments are available, the responsiveness and validity of the shorter questionnaires need to be tested by comparing the two versions. Finally, translation of an existing instrument into a new language requires re-examination of the measurement criteria (validity, reliability, and responsiveness). We anticipate a growing interest in measurement of clinical outcomes for patients with advanced lung diseases in the foreseeable future. We expect that current instruments may be modified and that, certainly, new tools will be developed in an attempt to improve the ability to measure dyspnea and HRQOL. Clearly, it is an ongoing process. PMID- 9329870 TI - Determination of disability for patients with advanced lung disease. AB - To conclude, impairment ratings differ among various diseases and compensation programs. It therefore is important to note, at the outset, which compensation program the patient is eligible for because the requirements of the different programs may vary. The physician report must clearly state an opinion using terminology understandable to lay people. It should include the diagnosis, whether the condition is work-related or not, the evidence of impairment, and the severity of impairment. It should also state whether the resultant disability is temporary or permanent. In the case of work-related diseases, apportionment should be addressed. Finally, patients with advanced lung disease may be totally disabled from certain types of employment and yet may be eligible for vocational training. It is appropriate to describe the types of jobs and work the patient physically can and cannot do. PMID- 9329871 TI - Assessment of operative risk for patients with advanced lung disease. AB - Increasingly, patients with advanced lung disease are being offered operative procedures. The assessment of the perioperative risk of these patients must include not only the assessment of their lung disease, but the assessment of the patient's cardiovascular disease, their age, and their other medical problems. Knowledge of the stress of particular surgical procedures is also of importance in risk assessment, and is addressed in this article. PMID- 9329872 TI - Anxiety and depression in advanced lung disease. AB - Anxiety, panic, and depression commonly complicate chronic airflow obstruction, and probably other forms of advanced lung disease as well. Despite the recent development of many new therapeutic options, these conditions remain under recognized and under-treated in this patient population. Under-diagnosis may result in part from the challenge of distinguishing between the somatic manifestations of psychiatric disease and the physical symptoms of severe respiratory dysfunction. Treatment relies on judicious pharmacotherapy and appropriate psychologic support. Serotonin selective reuptake inhibitors are particularly useful in the treatment of depression and panic, and may be helpful in controlling other forms of anxiety, as well. Cognitive behavioral therapy is an important adjunct in the management of anxiety. Electroconvulsive therapy should be considered for selected lung disease patients with refractory depression. PMID- 9329873 TI - Management of complications of glucocorticoid therapy. AB - Corticosteroids are commonly employed in the treatment of a wide variety of pulmonary disorders, including asthma, chronic obstructive pulmonary disease, pulmonary fibrosis, sarcoidosis, and pulmonary vasculitides; as well as in the lung transplant population. Frequently, steroid-induced complications contribute significantly to morbidity and diminished quality of life, often overshadowing the impact of the underlying lung disease. This article will discuss the more commonly encountered complications of corticosteroids in the context of the method of delivery, events occurring with chronic use, and events related to withdrawal. PMID- 9329874 TI - Pulmonary rehabilitation for patients with advanced lung disease. AB - Pulmonary rehabilitation has gradually become a cornerstone of treatment for patients with advanced lung disease. Although most of the data that has resulted in the acceptance of this therapeutic modality has been obtained from studies of patients with chronic obstructive pulmonary disease, the basic principles and tools are applicable to patients with many other limiting chronic diseases of the respiratory system. Because new therapeutic strategies, such as lung volume reduction surgery and lung transplantation, require well-conditioned patients, pulmonary rehabilitation is becoming a crucial component of the overall treatment strategy of many patients who heretofore were deemed untreatable. This article reviews the basic definitions, objectives, components, and outcomes of pulmonary rehabilitation in order to provide the reader with a practical and inclusive overview of the topic. PMID- 9329875 TI - Home oxygen therapy. AB - Home oxygen therapy has well-established benefits for patients with chronic obstructive pulmonary disease and resting hypoxemia. The indications for therapy have been clearly defined in the United States by the Health Care Financing Administration (HCFA), and these guidelines have been accepted by most third party payers. Controversies regarding the use of oxygen during sleep and exercise when daytime hypoxemia is not present have important therapeutic and financial implications. HCFA has recently proposed a substantial reduction in reimbursement for home oxygen which could have a major adverse effect if done on a global, across-the-board, basis. Some of the new technological developments in oxygen delivery systems are presented in this article. PMID- 9329876 TI - Nutritional support in advanced lung disease. The pulmonary cachexia syndrome. AB - Patients with advanced lung disease (ALD) demonstrate changes in body composition characteristically manifested by a progressive loss of body weight. The mechanisms of this pulmonary cachexia syndrome are multifactorial, and treatment must be comprehensive in nature. This article addresses our current knowledge regarding the relationship between nutrition and ALD. PMID- 9329877 TI - Options for long-term ventilatory support. AB - Long-term mechanical ventilator support for patients with chronic respiratory failure is becoming more common. This article reviews the common causes for chronic ventilator dependence, and offers an approach to weaning these patients from the ventilator. In addition, the details for preparing these patients for prolonged mechanical ventilation outside of the acute-care hospital setting are discussed. Appropriate education of the patient's caregivers is key to the success of long-term ventilatory support outside of the acute-care hospital. PMID- 9329878 TI - Surgical options for patients with advanced emphysema. AB - Since the early 1900s, a variety of operations have been suggested for emphysema but, with the exception of giant bullectomy, an option in only a small fraction of patients, none has proven effective. Data collected by a number of academic medical centers indicate that LVRS may ameliorate symptoms and improve pulmonary physiology, function, and quality of life in appropriately selected patients with emphysema. Accordingly, LVRS may provide an opportunity to intervene in a rapid, effective, and, possibly, cost-effective manner in a debilitating, chronic disease. That is an extraordinarily attractive proposition for both patients and physicians alike. But a number of questions remain: (1) What is the effect of LVRS compared with maximal medical therapy? (2) What is the duration of any beneficial effect of LVRS? (3) What is the best operative approach? (4) What patient characteristics predict good and bad outcomes? (5) What is the role of pre- and, possibly, postoperative pulmonary rehabilitation? (6) Does LVRS adversely affect the rate of loss of lung function over time, as some have suggested? (7) What is the cost of LVRS compared with standard medical therapy? (8) Can the procedure be performed safely in nontransplant centers? (9) What is the effect on disease-specific quality of life? (10) Does it affect mortality? A prospective, randomized controlled trial involving 18 selected centers will begin in the fall of 1997 under the sponsorship of the Health Care Financing Corporation (the administrators of Medicare) and the National Institutes of Health. We strongly support the creative, collaborative approach that has been taken by those two government agencies to stimulate this study. The need for controlled trials of new therapies cannot be overstated; only with such trials can the questions enumerated above be answered with certainty. PMID- 9329879 TI - Therapeutic options for severe pulmonary hypertension. AB - Pulmonary hypertension occurs as a consequence of numerous and varied conditions, all of which result in an elevation of pulmonary vascular resistance. Over the past decade, significant progress has been made in understanding the factors which contribute to the progressive nature of pulmonary vascular disease, and in identifying new treatments for pulmonary hypertension. The majority of these therapeutic options are pharmacologic, but for specific circumstances, surgical therapy may be a consideration. This article discusses nonspecific therapies for all patients with pulmonary hypertension, vasodilator therapy (including screening for vasodilator responsiveness, standard oral agents, and newer intravenous or inhalational therapies) and surgical options applicable to specific situations. PMID- 9329880 TI - Diagnosis and management of steroid-resistant asthma. AB - The term "steroid-resistant (SR) asthma" has been used to describe a group of asthmatics who demonstrate persistent airway obstruction and inflammation despite treatment with high doses of systemic glucocorticoids. There are at least two forms of SR asthma, that is, primary and acquired types. Type I SR asthma is acquired and is associated with abnormally reduced glucocorticoid receptor (GR) ligand and DNA binding affinity. Type II SR asthma is due to a primary GR binding abnormality. An important distinction between these two types of SR asthma is that the GR defect in Type I, but not Type II, SR asthma is reversible in culture and is sustained by incubation with combination IL-2 and IL-4. The treatment of these patients requires a systematic approach to rule out confounding factors, including triggers of immune activation, optimizing steroid therapy, and use of alternative strategies to inhibit airway inflammation. PMID- 9329881 TI - Lung transplantation. A disease-specific approach. AB - Lung transplantation has emerged as a viable option for the treatment of end stage disease attributable to a wide spectrum of primary disorders. Although many aspects of patient management are indifferent to the underlying indication, important differences related to timing of transplantation, selection of candidates, choice of procedure, and post-transplant complications exist among the various primary disease groups. Optimal utilization of transplantation for these challenging patient populations with advanced lung disease mandates a thorough appreciation of those differences. PMID- 9329882 TI - Advanced lung disease. Palliation and terminal care. AB - Considering that lung disease is the fourth leading cause of death in the United States, remarkably little has been written about palliative care for patients who die of respiratory disease. Because most such deaths are anticipated, palliative care should begin with advance medical planning, ideally in the form of a prescheduled meeting among the physician, the patient, and the patient's proxy for health affairs. Home hospice care should be considered when a patient with progressive lung disease is largely confined to the bedroom because of dyspnea. Medical attention during the terminal phase of a respiratory illness should focus on the experience of the patient. Common symptoms amenable to counseling and pharmacotherapy include dyspnea, pain, anxiety, insomnia, and depression. If initiated to no benefit, mechanical ventilation can be terminally withdrawn with the concurrence of the patient or family. The withdrawal process should be family centered, and followed by continued supportive care until the patient dies. PMID- 9329883 TI - Leprosy disabilities: the impact of multidrug therapy (MDT). PMID- 9329884 TI - A few more grains of melanin. PMID- 9329885 TI - The conference: an integral part of continuing medical education. PMID- 9329886 TI - Dermatology Online Journal: an Internet-based journal for dermatologists. PMID- 9329887 TI - Etiologic factors in Peyronie's disease. AB - BACKGROUND: The etiology of Peyonie's disease remains a mystery. An attempt is made to resolve this. MATERIALS AND METHODS: A total of 25 patients with Peyronie's disease were examined and investigated for the presence of various implicated etiologic factors. RESULTS: Of the 25 patients examined, two presented with ischemic heart disease, 10 with hypertension, 10 with asymptomatic hyperuricemia, 18 with hypercholesterolemia, two with fibrosing disorders, two with autoimmune disorders, and three with diabetes mellitus. None had any sexually transmitted disease. Seven of the 10 hypertensive patients had received beta-blockers. On comparing the prevalence of these factors between patients and the general population, a significant association was found between hypertension, hypercholesterolemia, hyperuricemia, and Peyronie's disease. CONCLUSIONS: Known factors, such as hypertension and hypercholesterolemia, lead to weakening of the vasculature and its rupture during coitus. This leads to the formation of a hematoma and its subsequent organization in predisposed individuals to fibrous plaques. The identification and treatment of these factors might prevent, secondarily, the development of Peyronie's disease. PMID- 9329888 TI - Primary cutaneous lymphomas: a study of 37 cases. AB - BACKGROUND: A retrospective clinical, histologic, and immunohistochemical study was performed in 37 cases of isolated primary cutaneous lymphoma (PCL) (22 B and 15 T phenotype). Patients with epidermotrophic infiltrate (mycosis fungoides and Sezary syndrome) were excluded. METHODS: Patients with PCL were selected according to strict criteria: isolated cutaneous involvement for at least 6 months and a negative exhaustive study of possible spread. Lesions were either limited to a single cutaneous region or were disseminated, involving at least two nonadjacent regions. The diagnosis was confirmed histologically, and an immunohistochemical study was performed. RESULTS: On the basis of the new Willemze classification for prognostic criteria, this study showed similarities between lymphomas of B and T phenotype in clinical features, therapeutic response, course, and overall prognosis. The clinical lesion was usually an erythematous nodule associated, or not, with an infiltrated layer and generally limited to a single cutaneous region. PCLs were highly sensitive to nonaggressive treatment, showing complete or more than 50% partial remission in all cases. CONCLUSIONS: The overall prognosis for these lymphomas was good, even for disseminated cutaneous forms. Patient survival at 48 months was 78% for T and 89% for B phenotype. In the latter group, the prognosis was comparable for CD30+ and CD30- T lymphomas; however, the course of PCL involved frequent cutaneous relapses, particularly with the disseminated forms, raising the problem of adjuvant treatment after complete remission was obtained. Extracutaneous involvement was rare, but always indicative of poor prognosis. PMID- 9329889 TI - Hair cycle-dependent expression of heat shock proteins in hair follicle epithelium. AB - BACKGROUND: Heat shock proteins (HSPs) have a physiologic function in unstressed cells, which is believed to include a role as a "molecular chaperone." The hair cycle is characterized by rhythmic tissue remodelling processes, and is an intriguing model for studying the relation between keratinocyte differentiation and HSP expression under physiologic circumstances. We have therefore studied, by immunofluorescence, the expression of selected HSPs during the murine hair cycle. METHODS: The association between hair follicle cycling and the expression of three selected HSPs (HSPs 27, 60, and 72) was examined by immunofluorescence, using the depilation-induced hair cycle of C57BL/6 mice. RESULTS: HSP expression was absent from telogen follicles, and was restricted predominantly to keratinocytes in the bulge and the cycling epithelial portion of the hair follicle during anagen and catagen. Immunoreactivity for HSPs 27, 60, and 72 in the hair bulb increased significantly during anagen VI and the catagen transformation of the follicle, and decreased again abruptly with completion of the catagen-telogen transformation. The expression pattern of HSPs 60 and 72 in situ was cytoplasmic, whereas that of HSP 27 was both cytoplasmic and nuclear. CONCLUSIONS: These observations suggest that the synthesis of HSPs by hair bulb keratinocytes is related to the anagen-catagen transformation of the follicle, possibly reflecting keratinocyte apoptosis and/or terminal differentiation in the regressing, cycling portion of the follicle. In addition, the rather proximal localization of HSP expression makes it unlikely that the HSPs examined interact with the more distally located intrafollicular gamma/delta T-cell receptor positive lymphocytes. PMID- 9329890 TI - Chemotherapy-induced acral erythema in leukemic patients: a report of 15 cases. AB - BACKGROUND: Chemotherapy-induced acral erythema is a distinct localized cutaneous response to certain systemic chemotherapeutic agents. METHODS: Between January 1990 and December 1994, from a total of 76 leukemic patients who have received combination chemotherapy consisting of cytosine arabinoside and anthracycline antibiotics, 15 patients developed chemotherapy-induced acral erythema. Fourteen of the patients had acute myelocytic leukemia, and one of them had chronic myelogenous leukemia in blast phase. Clinical features of these 15 patients have been analysed. Biopsy specimens obtained from eight of the patients were also evaluated for histopathologic alterations. RESULTS: The overall incidence of this reaction was found to be 19.7% in our group of patients receiving this chemotherapy protocol. The onset of reaction varied from the fourth to the seventeenth days of the chemotherapy and resolved within 2 weeks in most of the patients. Lesions appeared as well-defined erythema and edema involving the palmar surfaces in all of the patients. In nine of the patients the reaction recurred with subsequent chemotherapies. Scattered necrotic keratinocytes, vacuolar alterations of the basal layer, and mild to moderate perivascular lymphocytic infiltration in the dermis were the histopathologic findings observed in the biopsy specimens. CONCLUSIONS: Chemotherapy-induced acral erythema is a frequent reaction in patients who are receiving high-dose chemotherapy. For patients in whom this self-limited condition develops, reassurance is the mainstay of therapy. Awareness of this reaction is also important to be able to differentiate it from acute graft versus host disease in patients who receive bone marrow transplants. PMID- 9329891 TI - Cutaneous disseminated histoplasmosis in AIDS patients in south Florida. AB - BACKGROUND: Histoplasma capsulatum is a dimorphic pathogenic fungus endemic to the Mississippi and Ohio river valleys. In the immunocompetent it causes a self limited disease, but in the immunocompromised may lead to disseminated disease (disseminated histoplasmosis (DH)). It is one of the opportunistic infections which defines the acquired immunodeficiency syndrome (AIDS) and is rarely encountered outside endemic regions. METHODS: Clinical, laboratory, and histologic information concerning seven patients with DH and AIDS in South Florida was recorded. RESULTS: We report seven cases of DH with mucocutaneous lesions in patients infected with the human immunodeficiency virus (HIV). All patients had markedly depressed CD4 counts of less than 40 cells/mm3, and only two had traveled to endemic areas. Two out of the seven patients were diagnosed with HIV/AIDS at the time DH was identified. All of our patients had mucocutaneous lesions at the time of diagnosis, which clinically presented as a generalized papular eruption, ulcers, and erythematous scaly plaques. CONCLUSIONS: Even in non-endemic regions, HIV-positive patients presenting with fever, chills, weight loss, hepatosplenomegaly, anemia, cough, lymphadenopathy, and mucocutaneous lesions should have an early skin biopsy specimen taken for mycologic tissue culture and histopathologic evaluation for disseminated fungal infections. PMID- 9329892 TI - Tegafur-induced photosensitivity--evaluation of provocation by UVB irradiation. PMID- 9329893 TI - Subcutaneous nodules as initial metastatic sites in osteosarcoma. PMID- 9329894 TI - Paget's disease of the breast in a man with neurofibromatosis. PMID- 9329895 TI - Dermatomyositis and acquired ichthyosis as paraneoplastic manifestations of ovarian tumor. PMID- 9329896 TI - Disseminated recurrent papular B-cell pseudolymphoma. PMID- 9329897 TI - Sparfloxacin in the treatment of leprosy patients. AB - Seven men (age range, 20-33 years) with leprosy visited our dermatologic clinic at Yokohama City University Hospital (Table 1) and were entered into a trial of sparfloxacin (SPFX), 100-200 mg daily for up to 1 year. Five patients were Japanese Brazilians or Paraguayan from South America, and two patients were Filipinos. Examination procedures included a detailed medical history, pretreatment, clinical examinations, and body charting of the characteristic skin lesions, areas of anesthesia, and enlarged peripheral nerves. There were no deformities observed in any patient. All had presented with eruptions and neurologic problems. The diagnosis of leprosy and its type were determined by the above examination, skin smear, and histopathologic study, as well as Mitsuda reaction. The first patient came from the Philippines and presented to our clinic in 1993. There were many elevated erythemas with central healing on his whole body. Sensory nerves were impaired. His type was lepromatous (LL) leprosy. The second patient was followed for only 1 month because of a change in residence. The third and fourth patients were brothers. The fifth patient came from Paraguay. The sixth patient was from the Philippines. The seventh patient came to our clinic in April 1995. PMID- 9329898 TI - Basal cell carcinoma of trunk and extremities. AB - BACKGROUND: Basal cell carcinoma is the most common malignancy in Caucasians. Information about basal cell carcinoma in the Mexican population is scarce. OBJECTIVE: To determine the epidemiologic and clinical characteristics and treatment results of basal cell carcinoma located on the trunk and extremities of patients seen at the Instituto Nacional de Cancerologia of Mexico. METHODS: A retrospective study was performed of patients with confirmed diagnosis of basal cell carcinoma located on the trunk and extremities seen at the Instituto Nacional de Cancerologia of Mexico between 1966 and 1993. RESULTS: Ninety-one patients with basal cell carcinoma located on the trunk and extremities were found (6% of all patients with diagnosis of basal cell carcinoma). The median age was 64 years; 52% of the patients were women and 48% were men. A total of 119 basal cell carcinomas at these locations were diagnosed. The size of the skin tumor ranged from 0.3 to 22 cm (mean, 3.9 cm). Treatment results were evaluated in 62 patients (follow-up ranged from 24 to 240 months; mean, 80 months). Overall tumor control was accomplished in 95% of cases. Three patients died as a result of basal cell carcinoma. CONCLUSIONS: Basal cell carcinoma in the Mexican population is not as infrequent as previously thought, although it is less commonly located on the trunk and extremities than in Caucasians. PMID- 9329899 TI - Hansen's disease in Japan: a brief history. PMID- 9329901 TI - Pentoxifylline-induced thrombocytopenia. PMID- 9329900 TI - Pustular eruption in a malaria patient treated with chloroquine. PMID- 9329902 TI - Analyzing scanning microscopic study of contents of a giant comedo. PMID- 9329903 TI - Our man in Havana. PMID- 9329904 TI - Pyoderma gangrenosum of penile skin. PMID- 9329932 TI - Herpes simplex virus type 2 in the United States, 1976 to 1994. AB - BACKGROUND: Herpes simplex virus type 2 (HSV-2) infection is usually transmitted sexually and can cause recurrent, painful genital ulcers. In neonates the infection is potentially lethal. We investigated the seroprevalence and correlates of HSV-2 infection in the United States and identified changes in HSV 2 seroprevalence since the late 1970s. METHODS: Serum samples and questionnaire data were collected during the National Health and Nutrition Examination Surveys (NHANES) II (1976 to 1980) and III (1988 to 1994). HSV-2 antibody was assessed with an immunodot assay specific for glycoprotein gG-2 of HSV-2. RESULTS: From 1988 to 1994, the seroprevalence of HSV-2 in persons 12 years of age or older in the United States was 21.9 percent (95 percent confidence interval, 20.2 to 23.6 percent), corresponding to 45 million infected people in the noninstitutionalized civilian population. The seroprevalence was higher among women (25.6 percent) than men (17.8 percent) and higher among blacks (45.9 percent) than whites (17.6 percent). Less than 10 percent of all those who were seropositive reported a history of genital herpes infection. In a multivariate model, the independent predictors of HSV-2 seropositivity were female sex, black race or Mexican American ethnic background, older age, less education, poverty, cocaine use, and a greater lifetime number of sexual partners. As compared with the period from 1976 to 1980, the age-adjusted seroprevalence of HSV-2 rose 30 percent (95 percent confidence interval, 15.8 to 45.8 percent). The seroprevalence quintupled among white teenagers and doubled among whites in their twenties. Among blacks and older whites, the increases were smaller. CONCLUSIONS: Since the late 1970s, the prevalence of HSV-2 infection has increased by 30 percent, and HSV-2 is now detectable in roughly one of five persons 12 years of age or older nationwide. Improvements in the prevention of HSV-2 infection are needed, particularly since genital ulcers may facilitate the transmission of the human immunodeficiency virus. PMID- 9329933 TI - Acetaminophen toxicity in an urban county hospital. AB - BACKGROUND: The prevalence and characteristics of acetaminophen-associated liver injury in hospitalized patients are not well defined. METHODS: We identified patients hospitalized for excessive acetaminophen ingestion at an urban county hospital over a 40-month period (1992 to 1995) and reviewed their medical records to determine the incidence and clinical features of the ingestions and their outcomes. RESULTS: Of the 71 patients studied, 50 were classified as having taken acetaminophen during suicide attempts and 21 as having accidentally poisoned themselves while attempting to relieve pain. The suicidal patients had ingested almost twice as much acetaminophen as those in the accidental-overdose group (median, 20 vs. 12 g; P=0.009). Among the patients for whom data were available, 63 percent of those in the accidental-overdose group and 25 percent of those in the suicidal group had chronic alcohol abuse (P=0.009). The patients in the accidental-overdose group more often had severe liver necrosis (aminotransferase levels, >3500 IU per liter; 52 percent vs. 14 percent; P=0.002), and were more likely to have hepatic coma (33 percent vs. 6 percent, P=0.006). There were four deaths (19 percent) in the accidental-overdose group and one (2 percent) in the suicidal group (P=0.04). Five patients -- three in the accidental-overdose group and two in the suicidal group -- had ingested 4 g of acetaminophen or less. Acetaminophen ingestion accounted for 12 percent of all patients hospitalized with overdoses (71 of 589) and 40 percent of patients with acute liver failure (10 of 25) during the study period. CONCLUSIONS: In an urban county hospital, patients hospitalized with acetaminophen toxicity related to accidental misuse had higher rates of morbidity and mortality than those who attempted suicide, even though the latter had taken more acetaminophen. A higher frequency of chronic alcohol abuse among the patients with accidental overdoses may be one explanation. PMID- 9329934 TI - Images in clinical medicine. Orf skin ulcer. PMID- 9329935 TI - Anesthesiology. First of two parts. PMID- 9329936 TI - Evaluation and management of traumatic lacerations. PMID- 9329937 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 32-1997. A 43-year-old woman with rapidly changing pulmonary infiltrates and markedly increased intracranial pressure. PMID- 9329938 TI - Herpes simplex virus type 2--a persistent problem. PMID- 9329939 TI - Equivalence trials. PMID- 9329940 TI - National HIV case reporting for the United States. A defining moment in the history of the epidemic. PMID- 9329953 TI - A p47-phox pseudogene carries the most common mutation causing p47-phox- deficient chronic granulomatous disease. AB - The predominant genetic defect causing p47-phox-deficient chronic granulomatous disease (A47 degrees CGD) is a GT deletion (DeltaGT) at the beginning of exon 2. No explanation exists to account for the high incidence of this single mutation causing a rare disease in an unrelated, racially diverse population. In each of 34 consecutive unrelated normal individuals, both the normal and mutant DeltaGT sequences were present in genomic DNA, suggesting that a p47-phox related sequence carrying DeltaGT exists in the normal population. Screening of genomic bacteriophage and YAC libraries identified 13 p47-phox bacteriophage and 19 YAC clones. The GT deletion was found in 11 bacteriophage and 15 YAC clones. Only 5 exonic and 33 intronic differences distinguished all DeltaGT clones from all wild type clones. The most striking differences were a 30-bp deletion in intron 1 and a 20-bp duplication in intron 2. These results provide good evidence for the existence of at least one highly homologous p47-phox pseudogene containing the DeltaGT mutation. The p47-phox gene and pseudogene(s) colocalize to chromosome 7q11.23. This close linkage, together with the presence within each gene of multiple recombination hot spots, suggests that the predominance of the DeltaGT mutation in A47 degrees CGD is caused by recombination events between the wild type gene and the pseudogene(s). PMID- 9329954 TI - Myocardial insulin resistance in patients with syndrome X. AB - Insulin resistance is common in patients with angina pectoris, a positive exercise electrocardiogram, and normal coronary angiograms (syndrome X). It is still not known whether insulin resistance affects the cardiac muscle itself and, if so, whether insulin resistance involves myocardial hemodynamics and energy metabolism. We investigated hemodynamics as well as metabolite exchanges across the heart and the forearm in eight patients with syndrome X and eight control subjects during a baseline period after an overnight fast and during a hyperinsulinemic-euglycemic clamp. Myocardial hemodynamics and metabolism were studied at rest, during pace stress, and in the recovery period after pacing. Neither coronary sinus blood flow nor forearm blood flow differed between the groups before and during the clamp. Whole body insulin-stimulated glucose uptake was decreased in the patients (15.6+/-2.1 vs. 23.1+/-2.0 micromol x kg-1 x min 1). Insulin-stimulated glucose uptake in the forearm and the cardiac muscle was equally reduced in the patients (46+/-5 and 48+/-5%). Myocardial glucose uptake correlated with total arterial delivery in the control subjects (r = 0.63, P < 0.01), but not in patients (r = 0.22, P = 0.13). Carbohydrate and lipid oxidation was similar in the two groups at rest, and changes during the clamp were not different in control subjects and patients either at rest, during pacing, or in the recovery period. Patients with syndrome X exhibit myocardial insulin resistance, but cardiac energy metabolism remains unaffected. In patients with syndrome X, insulin-stimulated glucose uptake is independent from myocardial blood flow. PMID- 9329956 TI - Genetic identification of two major modifier loci of polycystic kidney disease progression in pcy mice. AB - Unlike the uniform disease progression in inbred animals, polycystic kidney disease (PKD) progression within human families can be highly variable. This may be due to environmental or genetic factors or both. To determine if PKD severity can be influenced by modifier genes, we carried out an intercross between DBA/2 pcy/pcy and Mus m. castaneous involving 3,105 6-wk-old F2 mice. Large differences in PKD severity were observed in this cross. In addition, 23/ 800 phenotypically normal mice were pcy/pcy genotypically. These results demonstrated that PKD progression in pcy/ pcy mice is a quantitative trait that is strongly modulated by modifier genes. Whole genome quantitative trait loci mapping of 114 selected pcy/pcy mice (68 with the mild PKD and 46 with severe PKD) identified two loci, MOP1 and MOP2 that strongly modulate PKD progression. MOP1 (max LOD score = 10.3 at D4Mit111) and MOP2 (max LOD score = 13.8 at D16Mit1) accounted for 36.7 and 46.8% of the phenotypic variance, respectively. Two-factor ANOVA of the phenotypes and genotypes of all 673 pcy/pcy mice from our cross indicated that MOP1 and MOP2 alleles regulate PKD progression in a complex additive manner. Characterization of these novel modifying loci may provide additional insights into the pathogenesis of polycystic kidney diseases. PMID- 9329955 TI - Prostaglandins I2 and E2 have a synergistic role in rescuing epithelial barrier function in porcine ileum. AB - Prostaglandins (PG) are cytoprotective for gastrointestinal epithelium, possibly because they enhance mucosal repair. The objective of the present studies was to assess the role of prostaglandins in intestinal repair. Intestinal mucosa from porcine ileum subjected to 1 h of ischemia was mounted in Ussing chambers. Recovery of normal transepithelial electrical resistance occurred within 2 h, and continued to increase for a further 2 h to a value twice that of control. The latter response was blocked by inhibition of prostaglandin synthesis, and restored by addition of both carbacyclin (an analog of PGI2) and PGE2, whereas the addition of each alone had little effect. Histologically, prostaglandins had no effect on epithelial restitution or villous contraction, indicating that elevations in transepithelial resistance were associated with increases in paracellular resistance. Furthermore, prostaglandin-stimulated elevations in resistance were inhibited with cytochalasin D, an agent known to stimulate cytoskeletal contraction. Synergistic elevations in transepithelial resistance, similar to those of carbacyclin and PGE2, were also noted after treatment with cAMP and A23187 (a calcium ionophore). We conclude that PGE2 and PGI2 have a synergistic role in restoration of intestinal barrier function by increasing intracellular cAMP and Ca2+, respectively, which in turn signal cytoskeletal mediated tight junction closure. PMID- 9329958 TI - Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer. AB - Both rheumatoid arthritis and animal models of autoimmune arthritis are characterized by hyperactivation of synovial cells and hyperplasia of the synovial membrane. The activated synovial cells produce inflammatory cytokines and degradative enzymes that lead to destruction of cartilage and bones. Effective treatment of arthritis may require elimination of most or all activated synovial cells. The death factor Fas/Apo-1 and its ligand (FasL) play pivotal roles in maintaining self-tolerance and immune privilege. Fas is expressed constitutively in most tissues, and is dramatically upregulated at the site of inflammation. In both rheumatoid arthritis and animal models of autoimmune arthritis, high levels of Fas are expressed on activated synovial cells and infiltrating leukocytes in the inflamed joints. Unlike Fas, however, the levels of FasL expressed in the arthritic joints are extremely low, and most activated synovial cells survive despite high levels of Fas expression. To upregulate FasL expression in the arthritic joints, we have generated a recombinant replication defective adenovirus carrying FasL gene; injection of the FasL virus into inflamed joints conferred high levels of FasL expression, induced apoptosis of synovial cells, and ameliorated collagen-induced arthritis in DBA/1 mice. The Fas ligand virus also inhibited production of interferon-gamma by collagen-specific T cells. Coadministration of Fas-immunoglobulin fusion protein with the Fas-ligand virus prevented these effects, demonstrating the specificity of the Fas-ligand virus. Thus, FasL gene transfer at the site of inflammation effectively ameliorates autoimmune disease. PMID- 9329957 TI - Hypercalcemia stimulates expression of intrarenal phospholipase A2 and prostaglandin H synthase-2 in rats. Role of angiotensin II AT1 receptors. AB - In chronic hypercalcemia, inhibition of thick ascending limb sodium chloride reabsorption is mediated by elevated intrarenal PGE2. The mechanisms and source of elevated PGE2 in hypercalcemia are not known. We determined the effect of hypercalcemia on intrarenal expression of cytosolic phospholipase A2 (cPLA2), prostaglandin H synthase-1 (PGHS-1), and prostaglandin H synthase-2 (PGHS-2), enzymes important in prostaglandin production. In rats fed dihydrotachysterol to induce hypercalcemia, Western blot analysis revealed significant upregulation of both cPLA2 and PGHS-2 in the kidney cortex and the inner and outer medulla. Immunofluorescence localized intrarenal cPLA2 and PGHS-2 to interstitial cells of the inner and outer medulla, and to macula densa and cortical thick ascending limbs in both control and hypercalcemic rats. Hypercalcemia had no effect on intrarenal expression of PGHS-1. To determine if AT1 angiotensin II receptor activation was involved in the stimulation of cPLA2 and PGHS-2 in hypercalcemia, we treated rats with the AT1 receptor antagonist, losartan. Losartan abolished the polydipsia associated with hypercalcemia, prevented the increase in cPLA2 protein in all regions of the kidney, and diminished PGHS-2 expression in the inner medulla. In addition, losartan completely prevented the increase in urinary PGE2 excretion in hypercalcemic rats. Intrarenal levels of angiotensin II were unchanged in hypercalcemia. These data indicate that hypercalcemia stimulates intrarenal cPLA2 and PGHS-2 protein expression. Our results further support a role for angiotensin II, acting on AT1 receptors, in mediating this stimulation. PMID- 9329959 TI - Cardiac compartment-specific overexpression of a modified retinoic acid receptor produces dilated cardiomyopathy and congestive heart failure in transgenic mice. AB - Retinoids play a critical role in cardiac morphogenesis. To examine the effects of excessive retinoid signaling on myocardial development, transgenic mice that overexpress a constitutively active retinoic acid receptor (RAR) controlled by either the alpha- or beta-myosin heavy chain (MyHC) promoter were generated. Animals carrying the alpha-MyHC-RAR transgene expressed RARs in embryonic atria and in adult atria and ventricles, but developed no signs of either malformations or disease. In contrast, beta-MyHC-RAR animals, where expression was activated in fetal ventricles, developed a dilated cardiomyopathy that varied in severity with transgene copy number. Characteristic postmortem lesions included biventricular chamber dilation and left atrial thrombosis; the incidence and severity of these lesions increased with increasing copy number. Transcript analyses showed that molecular markers of hypertrophy, alpha-skeletal actin, atrial natriuretic factor and beta-MyHC, were upregulated. Cardiac performance of transgenic hearts was evaluated using the isolated perfused working heart model as well as in vivo, by transthoracic M-mode echocardiography. Both analyses showed moderate to severe impairment of left ventricular function and reduced cardiac contractility. Thus, expression of a constitutively active RAR in developing atria and/ or in postnatal ventricles is relatively benign, while ventricular expression during gestation can lead to significant cardiac dysfunction. PMID- 9329960 TI - Upregulation of tumor necrosis factor-alpha gene by Epstein-Barr virus and activation of macrophages in Epstein-Barr virus-infected T cells in the pathogenesis of hemophagocytic syndrome. AB - A potentially fatal hemophagocytic syndrome has been noted in patients with malignant lymphomas, particularly in EBV-infected T cell lymphoma. Cytokines, such as interferon-gamma (IFN-gamma), TNF-alpha, and IL-1alpha, are elevated in patients' sera. To verify whether infection of T cells by EBV will upregulate specific cytokine genes and subsequently activate macrophages leading to hemophagocytic syndrome, we studied the transcripts of TNF-alpha, IFN-gamma, and IL-1alpha in EBV-infected and EBV-negative lymphoma tissues. By reverse transcription PCR analysis, transcripts of TNF-alpha were detected in 8 (57%) of 14 EBV-infected T cell lymphomas, higher than that detected in EBV-negative T cell lymphoma (one of six, 17%), EBV-positive B cell lymphoma (two of five, 40%) and EBV-negative B cell lymphomas (one of seven, 14%). Transcripts of IFN-gamma were consistently detected in T cell lymphoma and occasionally in B cell lymphoma, but were independent of EBV status. IL-1alpha expression was not detectable in any category. Consistent with these in vivo observations, in vitro EBV infection of T cell lymphoma lines caused upregulation of TNF-alpha gene, and increased secretion of TNF-alpha, but not IFN-gamma or IL-1alpha. Expression of TNF-alpha, IFN-gamma, and IL-1alpha was not changed by EBV infection of B cell lymphoma lines. To identify the specific cytokine(s) responsible for macrophage activation, culture supernatants from EBV-infected T cells were cocultured with a monocytic cell line U937 for 24 h. Enhanced phagocytosis and secretion of TNF alpha, IFN-gamma, and IL-1alpha by U937 cells were observed, and could be inhibited to a large extent by anti-TNF-alpha (70%), less effectively by anti-IFN gamma (31%), but almost completely by the combination of anti-TNF-alpha and anti IFN-gamma (85%). Taken together, the in vivo and in vitro observations suggest that infection of T cells by EBV selectively upregulates the TNF-alpha expression which, in combination with IFN-gamma and probably other cytokines, can activate macrophages. This study not only highlights a probable pathogenesis for virus associated hemophagocytic syndrome, but also suggests that anti-TNF-alpha will have therapeutic potential in the context of their fatal syndrome. PMID- 9329961 TI - Regulation of the gp80 and gp130 subunits of the IL-6 receptor by sex steroids in the murine bone marrow. AB - Both estrogen and androgen exert their antiosteoporotic effects, at least in part, by inhibiting IL-6 production, thereby suppressing osteoclastogenesis. Several observations, however, suggest that besides increased IL-6 production, sensitivity of the osteoclastogenic process to this cytokine is altered after ovariectomy. Based on this and evidence that the ligand-binding subunit of the IL 6 receptor (gp80) is a limiting factor for the actions of IL-6 on bone, we hypothesized that sex steroids regulate expression of the IL-6 receptor as well. We report that 17beta-estradiol or dihydrotestosterone in vitro decreased the abundance of the gp80 mRNA as well as the mRNA of the signal-transducing subunit of the IL-6 receptor (gp130) in cells of the bone marrow stromal/osteoblastic lineage, and also decreased gp130 protein levels. These effects did not require new protein synthesis. In contrast to sex steroids, parathyroid hormone stimulated gp130 expression; this effect was opposed by sex steroids. Consistent with these findings, ovariectomy in mice caused an increase in expression of gp80, gp130, and IL-6 mRNAs in ex vivo bone marrow cell cultures as determined by quantitative reverse transcription (RT)-PCR, and confirmed on an individual cell basis using in situ RT-PCR. The demonstration of increased expression of the IL-6 receptor after loss of sex steroids provides an explanation for why IL-6 is important for skeletal homeostasis in the sex steroid-deficient, but not replete, state. PMID- 9329962 TI - Overexpression of insulin-like growth factor-1 in mice protects from myocyte death after infarction, attenuating ventricular dilation, wall stress, and cardiac hypertrophy. AB - To determine whether IGF-1 opposes the stimulation of myocyte death in the surviving myocardium after infarction, transgenic mice overexpressing human IGF 1B in myocytes (FVB.Igf+/-) and wild-type littermates at 1.5 and 2.5 mo of age were subjected to coronary ligation and killed 7 d later. Myocardial infarction involved an average 50% of the left ventricle, and produced cardiac failure. In the region proximate to infarction, myocyte apoptosis increased 4. 2-fold and 2.1 fold in nontransgenics at 1.5 and 2.5 mo, respectively. Corresponding increases in myocyte necrosis were 1. 8-fold and 1.6-fold. In contrast, apoptotic and necrotic myocyte death did not increase in FVB.Igf+/- mice at either age after infarction. In 2.5-mo-old infarcted nontransgenics, functional impairment was associated with a 29% decrease in wall thickness, 43% increase in chamber diameter, and a 131% expansion in chamber volume. Conversely, the changes in wall thickness, chamber diameter, and cavitary volume were 41, 58, and 48% smaller in infarcted FVB.Igf+/- than in nontransgenics. The differential response to infarction of FVB.Igf+/- mice resulted in an attenuated increase in diastolic wall stress, cardiac weight, and left and right ventricular weight-to-body wt ratios. In conclusion, constitutive overexpression of IGF-1 prevented activation of cell death in the viable myocardium after infarction, limiting ventricular dilation, myocardial loading, and cardiac hypertrophy. PMID- 9329963 TI - A pharmacogenetic approach to blood pressure in Lyon hypertensive rats. A chromosome 2 locus influences the response to a calcium antagonist. AB - In a backcross population (n = 281) derived from a cross of the Lyon hypertensive rat with Lyon normotensive rat, we investigated whether genetic factors influence the acute cardiovascular responses to pharmacological modulation of the renin angiotensin system, the sympathetic nervous system, and the voltage-sensitive L type calcium channels. Using microsatellite markers, a quantitative trait locus was identified and mapped on rat chromosome 2 that specifically influences the systolic (peak LOD score 4.4) and diastolic (peak LOD score 4.1) blood pressure responses to administration of a dihydropyridine calcium antagonist, PY108-068. The locus accounted for 10.3 and 10.4% of the total variances in the systolic and diastolic responses to PY108-068, respectively. In marked contrast, the locus had no effect on either basal blood pressure or on the responses to acute administration of a ganglionic blocking agent, trimetaphan, or of an angiotensin II subtype 1 receptor antagonist, losartan. These findings provide strong direct support for the paradigm that genetic factors may influence the response to antihypertensive drugs and suggest that the heterogeneity seen in the responses to different antihypertensive agents in human essential hypertension may have a significant genetic determination. PMID- 9329964 TI - Insulin modulation of an endothelial nitric oxide component present in the alpha2 and beta-adrenergic responses in human forearm. AB - We explored in 51 normal subjects, distributed in various series of experiments, whether endothelium nitric oxide may play a role in insulin modulation of alpha2- and beta-adrenergic- evoked vascular responses. In particular, we examined the forearm blood flow response (FBF, ml.min-1.dl-1) to intrabrachial infusion of BHT 933 (0.5, 1, and 2 microg.min-1.dl-1) or isoproterenol (1, 3, and 6 ng. min-1.dl 1) in control conditions, during intrabrachial infusion of insulin alone (0.05 mU.kg-1.min-1) and associated with l-N-monomethylarginine (L-NMMA) (0.05 microg.min-1.dl-1), a nitric oxide synthase inhibitor. In control conditions both BHT-933 and isoproterenol induced a dose-dependent vascular response. Local hyperinsulinemia (deep venous plasma insulin 68.5+/-4 microU/ml) did not change basal FBF whereas attenuated BHT-933 vasoconstriction and enhanced isoproterenol vasodilation. L-NMMA reduced basal FBF and abolished the insulin effect on BHT 933 and isoproterenol response. To clarify whether a nitric oxide component is included in alpha2- and beta-adrenergic response and may be responsible for insulin vascular effect, we further examined BHT-933 and isoproterenol responses during nitric oxide inhibition. Interestingly, L-NMMA potentiated the BHT-933 vasoconstriction and attenuated the isoproterenol vasodilation and, in these conditions, insulin was no more able to exhibit its vascular effects. Finally, to rule out the possibility that the conteracting effect of L-NMMA may not be specifically related to insulin action, dose-response curves to phenylephrine (0.5, 1, and 2 microg.min-1.dl-1) or sodium nitroprusside (1, 2, and 4 microg.min 1.dl-1) were also performed. Both insulin and L-NMMA were unable to alter the phenylephrine-induced vasoconstriction and the sodium nitroprusside vasodilation. In conclusion, our data demonstrate an endothelial nitric oxide component in the alpha2- and beta-adrenergic vascular responses which is the target of the insulin vascular action. PMID- 9329965 TI - A non-MHC locus essential for autoimmune type I diabetes in the Komeda Diabetes Prone rat. AB - The Long-Evans Tokushima Lean (LETL) rat, characterized by rapid onset of insulin dependent (type I) diabetes mellitus (IDDM), no sex difference in the incidence of IDDM, autoimmune destruction of pancreatic beta cells, and no significant T cell lymphopenia, is a desirable animal model for human IDDM. We have established a diabetes-prone substrain of the LETL rat, named Komeda Diabetes-Prone (KDP) rat, showing a 100% development of moderate to severe insulitis within 220 d of age. The cumulative frequency of IDDM was 70% at 120 d of age, and reached 82% within 220 d of age. Here, we performed the first genome-wide scan for non-MHC IDDM susceptibility genes in this strain. The analysis of three crosses has led to the revelation of a major IDDM susceptibility gene, termed Iddm/kdp1, on rat chromosome (Chr) 11. Homozygosity for the KDP allele at this locus is shown to be essential for the development of moderate to severe insulitis and the onset of IDDM. Comparative mapping suggests that the homologues of Iddm/ kdp1 are located on human Chr 3 and mouse Chr 16 and would therefore be different from previously reported IDDM susceptibility genes. PMID- 9329966 TI - Targeted disruption of the beta-chemokine receptor CCR1 protects against pancreatitis-associated lung injury. AB - beta-Chemokines and their receptors mediate the trafficking and activation of a variety of leukocytes including the lymphocyte and macrophage. An array of no less than eight beta-chemokine receptors has been identified, four of which are capable of recognizing the chemokines MIP1alpha and RANTES. Genetic deletion of one of the MIP1alpha and RANTES receptors, CCR5, is associated with protection from infection with HIV-1 in humans, while deletion of the ligand MIP1alpha protects against Coxsackie virus-associated myocarditis. In this report we show that the deletion of another receptor for MIP1alpha and RANTES, the CCR1 receptor, is associated with protection from pulmonary inflammation secondary to acute pancreatitis in the mouse. The protection from lung injury is associated with decreased levels of TNF-alpha in a temporal sequence indicating that the activation of the CCR1 receptor is an early event in the systemic inflammatory response syndrome. PMID- 9329967 TI - Localization of distinct F2-isoprostanes in human atherosclerotic lesions. AB - F2-Isoprostanes are prostaglandin (PG) isomers formed in situ in cell membranes by peroxidation of arachidonic acid. 8-epi PGF2alpha and IPF2alpha-I are F2 isoprostanes produced in humans which circulate in plasma and are excreted in urine. Measurement of F2-isoprostanes may offer a sensitive, specific, and noninvasive method for measuring oxidant stress in clinical settings where reactive oxygen species are putatively involved. We determined whether isoprostanes were present in human atherosclerotic lesions, where lipid peroxidation is thought to occur in vivo. 8-epi PGF2alpha ranged from 1.310-3.450 pmol/micromol phospholipid in atherectomy specimens compared with 0.045-0.115 pmol/micromol phospholipid (P < 0.001) in vascular tissue devoid of atherosclerosis. Corresponding values of IPF2alpha-I were 5.6-13.8 vs. 0.16-0.44 pmol/micromol phospholipid (P < 0.001). Levels of the two isoprostanes in vascular tissue were highly correlated (r = 0.80, P < 0.0001). Immunohistochemical studies confirmed that foam cells adjacent to the lipid necrotic core of the plaque were markedly positive for 8-epi PGF2alpha. These cells were also reactive with anti-CD68, an epitope specific for human monocyte/macrophages. 8-epi PGF2alpha immunoreactivity was also detected in cells positive for anti-alpha-smooth muscle actin antibody, which specifically recognizes vascular smooth muscle cells. Our results indicate that 8-epi PGF2alpha and IPF2alpha-I, two distinct F2-isoprostanes and markers of oxidative stress in vivo, are present in human atherosclerotic plaque. Quantitation of these chemically stable products of lipid peroxidation in target tissues, as well as in biological fluids, may aid in the rational development of antioxidant drugs in humans. PMID- 9329968 TI - Inducible nitric oxide synthase suppresses the development of allograft arteriosclerosis. AB - In cardiac transplantation, chronic rejection takes the form of an occlusive vasculopathy. The mechanism underlying this disorder remains unclear. The purpose of this study was to investigate the role nitric oxide (NO) may play in the development of allograft arteriosclerosis. Rat aortic allografts from ACI donors to Wistar Furth recipients with a strong genetic disparity in both major and minor histocompatibility antigens were used for transplantation. Allografts collected at 28 d were found to have significant increases in both inducible NO synthase (iNOS) mRNA and protein as well as in intimal thickness when compared with isografts. Inhibiting NO production with an iNOS inhibitor increased the intimal thickening by 57.2%, indicating that NO suppresses the development of allograft arteriosclerosis. Next, we evaluated the effect of cyclosporine (CsA) on iNOS expression and allograft arteriosclerosis. CsA (10 mg/kg/d) suppressed the expression of iNOS in response to balloon-induced aortic injury. Similarly, CsA inhibited iNOS expression in the aortic allografts, associated with a 65% increase in intimal thickening. Finally, we investigated the effect of adenoviral mediated iNOS gene transfer on allograft arteriosclerosis. Transduction with iNOS using an adenoviral vector suppressed completely the development of allograft arteriosclerosis in both untreated recipients and recipients treated with CsA. These results suggest that the early immune-mediated upregulation in iNOS expression partially protects aortic allografts from the development of allograft arteriosclerosis, and that iNOS gene transfer strategies may prove useful in preventing the development of this otherwise untreatable disease process. PMID- 9329969 TI - Productive infection of dendritic cells by HIV-1 and their ability to capture virus are mediated through separate pathways. AB - There is substantial evidence that dendritic cells (DC) residing within epithelial surfaces (e.g., Langerhans cells) are the initial cells infected with HIV after mucosal exposure to virus. To study DC-HIV interactions in detail, we propagated Langerhans cell-like DC from cord blood CD34(+) cells and from adult blood plastic-adherent PBMC in the presence of cytokines (GM-CSF, IL-4, and/or TNF-alpha). DC pulsed overnight with HIVBaL or HIVIIIB were infected productively with both viral subtypes (as assessed by PCR, supernatant p24 protein levels, electron microscopy, and antibody staining). Productive infection could be blocked by anti-CD4 mAbs, RANTES (regulated upon activation, normal T cell expressed and secreted) (for HIVBaL), stromal cell-derived factor-1 (for HIVIIIB), or azidothymidine added during the HIV pulse, as well as by blocking DC proliferation. However, pulsing DC with HIV under these blocking conditions had no effect on the ability of DC to capture virus and transmit infection to cocultured antigen-stimulated CD4(+) T cells. Thus, we show by several criteria that (a) productive infection of DC and (b) the ability of DC to capture virus are mediated through separate pathways. We suggest that strategies designed to block mucosal transmission of HIV should consider interfering with both virus infection and virus capture by DC. PMID- 9329970 TI - Pertussis toxin-sensitive G proteins as mediators of the signal transduction pathways activated by cytomegalovirus infection of smooth muscle cells. AB - We demonstrated recently that the arachidonic acid (AA) cascade is involved in cytomegalovirus (CMV)-induced generation of reactive oxygen species (ROS) and the activation of nuclear factor (NF)-kappaB in human smooth muscle cells (SMCs). Since AA release from neutrophils is mediated by pertussis toxin (PTx)-sensitive guanine nucleotide-binding (G) proteins, we hypothesized by analogy that CMV stimulates ROS generation in SMCs and ultimately activates NF-kappaB via a PTx sensitive G protein-coupled pathway. Our first test of this hypothesis demonstrated that PTx blocked AA release induced by CMV infection of SMCs, as well as blocked the terminal products of this reaction, ROS generation and NF kappaB activation. More proximal components of the pathway were then examined. CMV infection increased phosphorylation and activity of cytosolic phospholipase A2 (cPLA2), an enzyme causing AA release; these effects were inhibited by PTx. CMV infection activated mitogen-activated protein (MAP) kinase, a key enzyme for cPLA2 phosphorylation, an effect also inhibited by PTx. Finally, inhibition of MAP kinase kinase (MAPKK), which phosphorylates and thereby activates MAP kinase, inhibited CMV-induced ROS generation. These data demonstrate that a PTx-sensitive G protein-dependent signaling pathway mediates cellular effects of CMV infection of SMCs. The downstream events include phosphorylation and activation of MAP kinase by MAPKK and subsequent phosphorylation and activation of cPLA2 (with its translocation to cell membranes), followed by stimulation of the AA cascade, which generates intracellular ROS and thereby activates NF-kappaB. PMID- 9329971 TI - Defective expression of gp180, a novel CD8 ligand on intestinal epithelial cells, in inflammatory bowel disease. AB - Previous studies support a role for intestinal epithelial cells (IEC) as antigen presenting cells in mucosal immune responses. T cells activated by IEC are CD8+, suppressor in function, and dependent upon CD8-associated p56lck activation. A 180-kD glycoprotein (gp180) recognized by mAbs B9 and L12 has been identified and shown to be important in CD8+ T cell activation by IEC. Since IEC derived from patients with inflammatory bowel disease (IBD) are incapable of activating CD8+ T cells, we asked whether this correlated with gp180 expression. While frozen sections of normal bowel revealed bright gp180 staining on all IEC, both inflamed and uninflamed ulcerative colitis (UC) specimens showed patchy staining. In Crohn's disease (CD), staining was faint to absent. Flow cytometry confirmed immunohistochemical data. The staining patterns correlated with the ability of IEC to activate CD8-associated p56lck. Normal IEC induced phosphorylation of p56lck in CD8alpha but not CD4+ transfectants. In contrast, both UC and CD IEC activated CD4 and, to a much lesser extent, CD8-associated p56lck. Thus, gp180 expression by IBD IEC appears to be altered, and correlates with a functional alteration of lck activation. This defect may reflect a more proximal event in the pathogenesis of IBD. PMID- 9329972 TI - Inhibition of tumor angiogenesis using a soluble receptor establishes a role for Tie2 in pathologic vascular growth. AB - Tie2 is a novel receptor tyrosine kinase that is expressed almost exclusively by vascular endothelium. Disruption of Tie2 function in transgenic mice resulted in embryonic lethality secondary to characteristic vascular defects; similar defects occurred after disruption of the Tie2 ligand. These findings indicate that the Tie2/Tie2 ligand pathway plays important roles during development of the embryonic vasculature. To determine whether the Tie2 pathway was involved in pathologic angiogenesis in adult tissues, a soluble form of the extracellular domain of murine Tie2 (ExTek.6His) was developed and used as a Tie2 inhibitor. After a single application of the ExTek.6His protein into a rat cutaneous window chamber, growth of a mammary tumor inside the chamber was reduced by > 75% (P < 0.005), and tumor vascular length density was reduced by 40% when compared with control-treated tumors (P < 0.01). In the rat cornea, ExTek.6His blocked angiogenesis stimulated by tumor cell conditioned media. ExTek.6His protein did not affect the viability of cultured tumor cells, indicating that the antitumor effect of ExTek.6His was due to the inhibition of tumor angiogenesis. These data demonstrate a role for the Tie2 pathway in pathologic angiogenesis, suggesting that targeting this pathway may yield effective antiangiogenic agents for treatment of cancer and other angiogenic diseases. PMID- 9329973 TI - A synthetic peptide derived from the sequence of a type I collagen receptor inhibits type I collagen-mediated platelet aggregation. AB - A synthetic peptide-1, an 18 amino acid residue peptide derived from a hydrophilic domain of a cloned platelet type I collagen receptor, was used to study the role of the receptor on types I and III collagen-induced platelet aggregation and the release of ATP. The peptide inhibits the type I, but not the type III, collagen-induced platelet aggregation and the release of ATP in a dose dependent manner. The [125I]peptide-1 specifically binds to type I collagen coated microtiter wells in a dose-dependent manner (with Kd = 10 nM). The binding of [125I]peptide-1 can be inhibited by an excess of unlabeled peptide-1 suggesting that the binding is specific. The labeled peptide-1 does not bind to type III collagen-coated microtiter wells. Results from an enzyme-linked immunosorbent assay show that the peptide reacts with the poly- and monoclonal antibodies raised against the purified platelet type I collagen receptor (Mr 65 kD). The peptide also inhibits the adhesion of platelets on type I collagen matrix and rabbit aortic segments in a dose-dependent manner. These results suggest that the reactive site of the platelet receptor for type I collagen resides in this portion of the molecule. PMID- 9329974 TI - Neutrophil accumulation on activated, surface-adherent platelets in flow is mediated by interaction of Mac-1 with fibrinogen bound to alphaIIbbeta3 and stimulated by platelet-activating factor. AB - We have studied the pathways that lead to arrest and firm adhesion of rolling PMN on activated, surface-adherent platelets. Stable arrest and adhesion strengthening of PMN on thrombin-stimulated, surface-adherent platelets in flow required distinct Ca2+- and Mg2+-dependent regions of Mac-1 (alphaMbeta2), and involved interactions of Mac-1 with fibrinogen, which was bound to platelets via alphaIIbbeta3. Mac-1 also bound to other unidentified ligands on platelets, which were not intracellular adhesion molecule-2 (ICAM-2), heparin, or heparan-sulfate proteoglycans. This was shown by inhibition with mAbs or peptides, by treatment of platelets with heparitinase, and by using platelets with defective alphaIIbbeta3 from a patient with Glanzmann thrombasthenia. Tethering of PMN on platelet ICAM-2 via LFA-1 (alphaLbeta2) was observed, which may facilitate the transition between rolling on selectins and Mac-1-dependent arrest. Arrest and adhesion strengthening was pertussis toxin sensitive and in flow was mainly induced by platelet-activating factor but not through activation of the chemokine receptor CXCR2. In stasis, spreading occurred and the CXCR2 contributed to firm adhesion. PMID- 9329975 TI - Pancreatic beta cells are important targets for the diabetogenic effects of glucocorticoids. AB - Abnormalities contributing to the pathogenesis of non-insulin-dependent diabetes mellitus include impaired beta cell function, peripheral insulin resistance, and increased hepatic glucose production. Glucocorticoids are diabetogenic hormones because they decrease glucose uptake and increase hepatic glucose production. In addition, they may directly inhibit insulin release. To evaluate that possible role of glucocorticoids in beta cell function independent of their other effects, transgenic mice with an increased glucocorticoid sensitivity restricted to their beta cells were generated by overexpressing the glucocorticoid receptor (GR) under the control of the insulin promoter. Intravenous glucose tolerance tests showed that the GR transgenic mice had normal fasting and postabsorptive blood glucose levels but exhibited a reduced glucose tolerance compared with their control littermates. Measurement of plasma insulin levels 5 min after intravenous glucose load demonstrated a dramatic decrease in acute insulin response in the GR transgenic mice. These results show that glucocorticoids directly inhibit insulin release in vivo and identify the pancreatic beta cell as an important target for the diabetogenic action of glucocorticoids. PMID- 9329976 TI - Host defense against systemic infection with Streptococcus pneumoniae is impaired in E-, P-, and E-/P-selectin-deficient mice. AB - Endothelial selectins mediate rolling of leukocytes on endothelium, a crucial step for leukocyte firm adhesion and emigration into sites of tissue injury and infection. To characterize the role of the endothelial selectins during bacterial sepsis in vivo, Streptococcus pneumoniae (1-10 x 10(6) colony-forming units) was inoculated intraperitoneally into wild-type mice and mice with E-, P-, or E-/P selectin deficiencies. Mice were followed 10 d for morbidity, survival, clearance of bacteremia, and leukocyte migration to the peritoneal cavity and organs 48 h after infection. All selectin-deficient mice showed a more pronounced morbidity, a significantly higher mortality associated with persistent bacteremia, and a higher bacterial load when compared with wild-type mice. These differences were most remarkable in the E-selectin-deficient mice, which showed the highest rate of mortality and bacteremia (P 10% weight loss. Before hrGH, rates of skeletal muscle protein synthesis, measured with l-[2H5]phenylalanine, were the same in controls and in all stages of disease. Rates of myofibrillar protein degradation, however, assessed from urinary excretion of 3-methyl histidine, were higher in AIDS and AIDS wasting than in HIV+ or healthy individuals. The group with weight loss had significantly higher TNFalpha levels but not higher HIV viral loads. Muscle function, as determined by isokinetic knee extension and shoulder flexion, was significantly higher in controls than all infected individuals. After GH, rates of protein synthesis were stimulated 27% in controls, with a smaller increase (11%) in HIV+, and a significant depression (42%) in AIDS with weight loss, despite fourfold elevation in insulin-like growth factor-I in all groups. There was a significant correlation of hrGH-induced changes in muscle protein synthesis with severity of disease (P = 0.002). The results indicate increased basal muscle protein degradation and decreased responsiveness of muscle protein synthesis to GH in the later stages of disease. PMID- 9329982 TI - Investigation of the influence of maternal infection with Wuchereria bancrofti on the humoral and cellular responses of neonates to filarial antigens. AB - Epidemiological data indicate that maternal filarial infection might be associated with increased susceptibility to filarial infection in offspring. To examine the influence of maternal infection on development of antifilarial immunity in neonates, paired cord and maternal sera and mononuclear cells were collected in an area where Wuchereria bancrofti infection is endemic. Anti filarial humoral responses (IgG, IgM and IgE) non-parasite-specific humoral responses (total IgE), proliferation induced by filarial antigen and production of cytokines (interleukin-2, interleukin-4 and interferon-gamma) were all monitored. Few cord serum samples had detectable antifilarial IgM of IgE and neither these responses nor total IgE levels differed as a function of maternal infection status. Of cord-blood mononuclear cells assayed, a relatively small proportion exhibited reactivity to filarial antigens. Based on these limited responses to filarial antigens, few neonates display evidence of in-utero sensitization to filarial antigens. PMID- 9329984 TI - Epidemiological and clinical studies on an outbreak of trichinosis in central China. AB - An outbreak of trichinosis occurred in the city of Zhengzhou, central China, between December 1995 and February 1996, affecting 85 of the administrative units into which the city is split. Of 297 subjects from eight of the affected units, 54% were seropositive for Trichinella and 41% had symptoms consistent with acute trichinosis. Of the 490 subjects who had eaten at one particular dumpling restaurant 1-5 weeks before the outbreak and who were traced, 291 (59%) were seropositive and 212 (43%) had been or were ill. MOst of the infections were in manual workers, cadres and merchants aged 20-49 years. Most of those who had been infected failed to develop gastro-intestinal symptoms or a cutaneous rash. Eyelid oedema was only seen in the early stages of the infection, the main clinical manifestations being fever of long duration of tiredness. Surprisingly, six cases had no marked symptoms after repeated infection. Eosinophilia (eosinophils > 7% of leucocytes) was noted in 71 (55%) of the 130 cases in which blood cells were counted. When 212 sera were tested for antibodies to Trichinella, seropositivities were found to increase from 89.1% (IFAT) of 87.7% (microprecipitation test) at presentation to 100% (both tests) 1 week after treatment with albendazole. All those treated were cured. The outbreak was one of the most extensive, single-source outbreaks ever recorded in China, probably with > 600 infections and > 300 clinical cases. The entire episode was attributed to the ingestion of undercooked pork dumplings at one restaurant. PMID- 9329980 TI - Widely dispersed p53 mutation in respiratory epithelium. A novel mechanism for field carcinogenesis. AB - Individuals with one aerodigestive tract malignancy have a high incidence of second primary aerodigestive tumors. The mechanism for this field effect has not been determined. We studied an individual with widespread dysplastic changes in the respiratory epithelium but no overt carcinoma. The entire tracheobronchial tree obtained at autopsy was embedded in paraffin, and bronchial epithelial cells were isolated by microdissection. DNA extracted from the microdissected cells was analyzed for point mutations in the p53 tumor suppressor gene. A single, identical point mutation consisting of a G:C to T:A transversion in codon 245 was identified in bronchial epithelium from 7 of 10 sites in both lungs. Epithelium at sites containing the p53 mutation was morphologically abnormal, exhibiting squamous metaplasia and mild to moderate atypia. No invasive tumor was found in the tracheobronchial tree or any other location. Cells from peripheral blood, kidney, liver, and lymph node exhibited no abnormality in the p53 gene. The widespread presence of a single somatic p53 point mutation in the bronchi of a smoker suggests that a single progenitor bronchial epithelial clone may expand to populate broad areas of the bronchial mucosa-a novel mechanism for field carcinogenesis in the respiratory epithelium that may be of importance in assessing individuals for risk of a second primary tumor as well as in devising effective strategies for chemoprevention of lung cancer. PMID- 9329983 TI - Successful control of onchocerciasis vectors in San Vicente Pacaya, Guatemala, 1984-1989. AB - Between 1984 and 1989, the onchocerciasis-vector control zone on the pilot area of San Vicente Pacaya, Guatemala, which had been subject to experimental control since 1979, was extended from 91.3 to 148.6 km2. Temephos was used as a larvicide against Simulium ochraceum s.l. the target species. As a new strategy, only breeding sites with water discharges of 0.1-10 litres/s were treated, every 2 weeks. This approach provided a substantial reduction in effort, number of treated sites, time and cost. To assess the effect of the temephos, nine sites were selected in which standardized collections of adult Simulium were made twice a month, by human bait. There was an obvious difference between the pre- and post control mean densities of flies at each site. In the northern area, which includes the Lavaderos, Barretal, Colina and Rodeo sites, the biting density in 1979, before treatment, varied between 10 and 64 flies/man-hour (FMH). Four years later, this had been reduced to 0.1-3.2 FMH, and by the end of the present study, in 1989, the mean density was zero FMH. In the southern area, which lies south east of Lavaderos (and includes Guachipilin, Ingerto, Pena Blanca and Sierra Morena), the density during the pre-control phase was 24 FMH at one of the two sites investigated at the time and 39.3 FMH at the other. It fell to 0.1-0.5 FMH after 5 years of control and to zero (three sites) or close to zero (< or = 0.5 FMH; one site) for the last 4 years of the present study. To assess the effect of vector control on onchocerciasis prevalence and incidence, 1280 residents from six endemic communities, out of 12,000 permanent inhabitants, were examined. In Santa Cruz, Patrocinio and Los Rios, the prevalence of skin microfilariae in the subjects from each community fell from 8.1%-37.8% during the pretreatment, base line period to 0.0%-31.5% when the study foci were totally integrated into the vector-control operation following treatment. Incidence among children (aged < or = 9 years) varied from 0%-25% for the period 1982-1984 but, thereafter, not a single case appeared in four of the six study communities (Santa Cruz, Patrocinio, Los Rios and Berlin). Incidence in Guachipilin did not decline appreciably, probably because of human migration into the area from other onchocerciasis foci. The prevalence of nodules followed a similar trend to those of the prevalence and incidence of skin microfilariae, falling from 9.1%-45.0% pre-control to 1.8%-14.3% 10 years later. PMID- 9329985 TI - Post-hepatitis aplastic anaemia: causal link to viral hepatitis A to G? PMID- 9329986 TI - Hydrocarbon variations/discrimination between two strains of Anopheles albimanus Wied from El Salvador. PMID- 9329988 TI - Progress in the map-based cloning of the Anopheles gambiae genes responsible for the encapsulation of malarial parasites. AB - A genetically selected strain of the mosquito Anopheles gambiae, the major vector of malaria in sub-Saharan Africa, is able to encapsulate and kill Plasmodium ookinetes after they have penetrated the midgut cells and come to rest between the midgut epithelial cells and the surrounding basal lamina. The genetic basis of this phenotype has now been examined by high-resolution mapping using microsatellite loci. Results of this mapping indicate that three genes contribute to this phenotype, with one gene on the left arm of chromosome 2 accounting for the most of the effect. These genes, called Pen1, Pen2, and Pen3 (for Plasmodium encapsulation genes 1, 2 and 3) have also been physically localized to relatively small and well defined regions of the polytene chromosome complement. Strategies for cloning these genes by genetic and physical mapping methods are discussed. PMID- 9329987 TI - Malarial pigment (haemozoin): a very active 'inert' substance. AB - Malarial pigment (haemozoin; HZ) is generally considered to be a non-toxic, high molecular-weight storage form of undigested, toxic, host-haemoglobin haem. The available information on HZ indicates that it is a very heterozygous material. Its exact structure, in terms of constituent proteins (remnants of host globin v. parasite proteins), the type of linkage between the haem moieties (mu-oxo haem dimers further aggregated by non-covalent hydrophobic bonds v. mutually independent haematin monomers), iron status in the haem (penta-co-ordinated, high spin ferriprotoporphyrin IX v. esa-co-ordinated, low-spin ferriprotoporphyrin IX), and compositions (beta-haematin-like structure without functionally relevant proteins or other constituents v. a ferriprotoporphyrin-IX core with aggregated proteins and phospholipids of host and parasite origin) remains a subject of controversy. When investigated by macrophages, HZ is not inert but affects a number of functional parameters. Crude pigment, as present in infected erythrocytes and shed after schizont rupture, may be considered the 'natural diet' ingested by macrophages in infected blood. It is a powerful source of radicals that may generate lipoperoxides and derived, toxic hydroxyaldehydes such as 4-hydroxynonenal (HNE). High concentrations of HNE, which have been detected in HZ-fed macrophages, inhibit protein kinase C (PKC). Complexes between HNE and PKC have also been detected in immunoprecipitated PKC from HZ-fed macrophages. HNE-mediated inhibition of PKC (and of other, as yet unidentified enzymes and processes) may explain HZ-mediated effects. HZ-mediated inhibition of NADPH oxidase, the enzyme responsible for oxidative bursts, may only be partially explained by PKC inhibition. As Hz-laden human and murine macrophages produce increased amounts of tumour necrosis factor-alpha, interleukins 1 and 6, and macrophage inflammatory proteins 1 alpha and 1 beta, HZ-macrophage interactions may contribute to the cytokine-mediated manifestations of malaria. PMID- 9329989 TI - What's new in malaria control? AB - Malaria remains a significant health problem in many tropical areas but the main impact of the infection is felt in sub-Saharan Africa, where malaria continues to cause many deaths and much morbidity. Recently, several new initiatives to improve malaria control in Africa have been started. Control is difficult to achieve in areas with very high levels of transmission but something can be accomplished with existing tools, which include provision of good treatment facilities, chemoprophylaxis in pregnancy and use of insecticide-treated materials. In some areas, especially on the fringes of sub-Saharan Africa, households spraying may also have a role. Use of insecticide-treated materials has had a major impact on child mortality in several countries in Africa, although there are concerns that the dramatic effects achieved initially may not be sustained. A major constraint on malaria control in Africa has been poor organization of malaria-control programmes. New approaches that combine elements of vertical and of horizontal control systems are being tried. Malaria control, even when carried out efficiently and cost-effectively, is relatively expensive. New ways of financing malaria control in the countries where it is needed most must be found. PMID- 9329990 TI - The malarial fever response--pathogenesis, polymorphism and prospects for intervention. AB - It is estimated that over 200 million people each year suffer debilitating attacks of malarial fever, and roughly 2 million of these episodes are fatal. The fever is caused by tumour necrosis factor (TNF) and other pyrogenic cytokines that are released by the host immune system response to products of schizont rupture. TNF has anti-parasitic properties but excessive TNF production is thought to play an important role in the pathogenesis of cerebral malaria. This review summarizes recent attempts to achieve molecular characterization of the parasite components that stimulate the host TNF response, and to define the host and parasite factors that affect the level of TNF production. Of particular interest are host polymorphisms that may regulate TNF gene expression, and naturally acquired antibodies that prevent the parasite from inducing TNF, both of which correlate with the clinical severity of infection. Our understanding of these processes, which are potentially of considerable therapeutic relevance, remains very limited at both the molecular and the epidemiological level. PMID- 9329991 TI - Protein trafficking in the Plasmodium-falciparum-infected erythrocyte--from models to mechanisms. AB - Plasmodium falciparum is a eukaryotic single cell which invades human erythrocytes. Within the host cell, the parasite is surrounded by the membrane of the parastiophorous vacuole. Parasite proteins are secreted either into the vacuolar space or are exported, by an as yet unknown mechanism, across the vacuolar membrane into erythrocyte cytoplasm. In recent years, several groups have devised experimental approaches to follow the transport pathways of proteins from the parasite into the host cell. The concepts underlying these approaches and the peculiarities of the transport pathways are discussed and compared with protein transport in higher eukaryotes. PMID- 9329992 TI - PfEMP1, polymorphism and pathogenesis. AB - The virulence of Plasmodium falciparum relative to the other species of malarial parasite which infect humans is thought to be due to this parasite's ability to adhere to endothelial cells lining small blood vessels and, in some cases, to its ability to form rosettes with uninfected erythrocytes. The latter phenotype has been found more frequently in cases of severe disease. The former property means that only the younger, asexual, intra-erythrocytic forms circulate whereas the more mature developmental stages are sequestered in the vasculature of a variety of organs. When large numbers of parasites accumulate in a vulnerable target organ such as the brain, the the life-threatening condition of cerebral malaria may result. While the factors that control whether or not cerebral malaria develops are not clearly defined, one crucial determinant my be the endothelial receptors utilised by the infecting isolate. Many such receptors have been identified, including CD36, thrombospondin, ICAM-1, VCAM, E-selectin and chondroitin-4-sulphate. The results of laboratory, field, post-mortem and direct receptor-binding studies indicate that, of the receptors currently identified, ICAM-1 binding is more likely to be associated with the development of cerebral malaria. The molecule expressed on the surface of the infected erythrocyte which mediates adherence to endothelium belongs to a large family of clonally variable antigens encoded by the var genes. The evidence for this conclusion and progress in defining the regions of var-gene products responsible to receptor-specific binding are discussed. Finally, the organization of the var genes within and between parasites is discussed in relation to the evolution of the var-gene family and its functions of antigenic variation and endothelial adhesion. PMID- 9329993 TI - Haematin (haem) polymerization and its inhibition by quinoline antimalarials. AB - Haematin (ferriprotoporphyrin IX) is released from haemoglobin during its degradation in the malarial parasites' food vacuole and is detoxified by its polymerization into a form of beta-haematin called haemozoin, or malarial pigment. This process is protein independent in vitro. Quinoline antimalarial blood schizonticides accumulate in the food vacuole and may inhibit haematin polymerization by binding to haematin and preventing its incorporation into the growing haemozoin chain. Drug resistance to quinolines is thought to be due to reduced accumulation of the drug in the food vacuole. As some quinolines overcome this resistance, quinolines, as a class, remain a potential source of future antimalarial drugs. PMID- 9329994 TI - Management of infectious diarrhoea in childhood. PMID- 9329995 TI - Surgical treatment of parathyroid disease. PMID- 9329996 TI - Cross-cultural competencies in the psychiatric assessment. AB - We live in an increasingly multicultural society and thus the cross-cultural application of assessment procedures is likely to become more common. However, little attention has been directed to the limitations of such unconditional application of what are essentially ethnocentric procedures. This article outlines the limitations of the psychiatric assessment when applied across cultures and presents the template for a culture sensitive assessment. PMID- 9329997 TI - Arterial cannulation: how to do it. AB - The use of arterial lines is now common in the care of critically ill patients. Intra-arterial cannulation with continuous blood pressure transduction and display remains the accepted standard for comprehensive arterial blood pressure monitoring. This article will illustrate a technique for percutaneous radial artery cannulation as well as outlining the indications, contraindications and possible complications of arterial cannulation. PMID- 9329998 TI - Neurological cases for MRCP: I. PMID- 9329999 TI - Decision-making in surgery: the management of gastric carcinoma. PMID- 9330000 TI - Endometrial resection. PMID- 9330001 TI - Listening to the patient: endometrial resection. PMID- 9330002 TI - Laparoscopic hysterectomy. PMID- 9330003 TI - How to do it in surgery: laparoscopic sacral colpopexy. PMID- 9330004 TI - Alcohol and the liver. AB - Alcohol remains the most common cause of cirrhosis in the Western world. This article reviews the increased understanding of the mechanisms by which alcohol damages the liver, why individuals differ in their susceptibility to alcohol, current treatments available and those which may have a role to play in the future. PMID- 9330005 TI - The management of lumps in the neck. AB - The many diagnostic problems associated with neck lumps demand a systematized and team approach to their management. Use of current imaging technology, effective examination of the upper aerodigestive tract and minimal surgical techniques for tissue diagnosis are discussed. PMID- 9330006 TI - Year to end budget forecasting: income and expenditure. AB - The introduction of devolved management structures within the NHS has exposed many clinical staff to the disciplines of financial management. This short article looks at the principles behind the preparation of budgets, the purpose of the income and expenditure account and the tools used to monitor expenditure at directorate and department level within a hospital. PMID- 9330007 TI - The management of shoulder pain. PMID- 9330008 TI - Quality and clinical audit. PMID- 9330009 TI - Meningococcal septicaemia. PMID- 9330010 TI - Attention deficit hyperactivity disorder or hyperkinetic disorder in adults. PMID- 9330011 TI - The Mental Health (Patients in the Community) Act 1995. A clinical analysis. PMID- 9330012 TI - Bereavement and grief in adults with learning disabilities. AB - BACKGROUND: This paper reports the results of the first systematic study of the reaction of people with learning disabilities to bereavement. METHOD: A sample of 50 parent-bereaved people with learning disabilities was compared with a matched control group of 50 non-bereaved people. A semi-structured bereavement questionnaire was used along with the following instruments: the Aberrant Behaviour Checklist (ABC), the Psychopathology Instrument for Mentally Retarded Adults (PIMRA) and the Life Events Checklist. RESULTS: Highly significant differences are demonstrated between bereaved and non-bereaved samples on both the total scores and most of the subscores of the ABC and PIMRA. Staff and carers did not usually attribute behaviour problems to the bereavement and its concomitant life events, nor was there a recognition of psychopathology due to bereavement. CONCLUSIONS: The impact in terms of psychiatric and behavioural morbidity of loss of a parent, with its concomitant life events, in adults with learning disabilities has been underestimated. PMID- 9330013 TI - Autistic traits in adults with learning disabilities. AB - BACKGROUND: Although many adults with learning disabilities show features of autistic syndrome, there have been very few population-based studies. We explored the prevalence of autistic traits and their association with maladaptive behaviours in a geographically defined population of adults with learning disabilities. METHOD: The carers of 2201 adults with learning disabilities were interviewed, and information was sought concerning aspects of their behaviour and ability. Individuals were scored according to the number of core autistic traits displayed. The prevalence of autistic traits was examined in respect of aspects of behaviour and ability. RESULTS: Autistic traits were common among adults with learning disabilities. Those with a higher number of autistic traits were more likely to be profoundly learning disabled and demonstrate a wide range of challenging behaviours. CONCLUSION: Many adults with learning disabilities demonstrate autistic traits. The relationship of autistic traits with challenging behaviour has major implications in service planning and delivery. PMID- 9330014 TI - An open trial of clozapine in neuroleptic-resistant childhood-onset schizophrenia. AB - BACKGROUND: Studies performed with schizophrenic adults who were resistant to classical neuroleptics showed improvement in 30% of the patients when treated with clozapine. Very early onset schizophrenic patients benefit only partially from conventional antipsychotics and are at increased risk of developing extrapyramidal symptoms; clozapine may offer an alternative treatment for these patients. METHODS: Eleven neuroleptic-resistant children (< 13 years) with schizophrenia were treated with clozapine. Improvement was monitored during the first 16 weeks using the Brief Psychiatric Rating Scale, Positive and Negative Syndrome Scale and Clinical Global Impression. The mean clozapine dosage was 227.3 (s.d. 34.4 mg/day at the end of the 16 weeks. RESULTS: There was an overall statistically significant reduction in all parameters, especially positive symptoms, implying a favourable outcome. Most of the improvement occurred during the first 6 to 8 weeks. The major side-effects were somnolence and drooling (no agranulocytosis). CONCLUSION: Clozapine may be a promising drug for the treatment of resistant childhood-onset schizophrenia. PMID- 9330015 TI - A controlled family study of late-onset non-affective psychosis (late paraphrenia). AB - BACKGROUND: The relationship between those schizophrenia-like conditions that have their onset in late life and early-onset schizophrenia is unclear. Very few family history studies of patients with late-onset psychosis have been reported, and it is not known whether their relatives have an increased risk of psychosis. METHOD: Information was collected on the psychiatric morbidity of 269 first degree relatives of patients with schizophrenia or delusional disorder with an onset after the age of 60 (late paraphrenia), and 272 first-degree relatives of healthy elderly control subjects, using a research diagnostic instrument. RESULTS: With a narrow age range (15-50 years) at risk, the estimated lifetime risk of schizophrenia was 1.3% in the relatives of both cases and controls. With a wider age range (15-90 years) at risk, estimated lifetime risk of schizophrenia was 2.3% for the relatives of cases and 2.2% for the relatives of controls. However, depression was significantly more common among the relatives of cases than controls. CONCLUSIONS: Those schizophrenia-like psychoses with onset in late life are not genetically associated with schizophrenia. PMID- 9330016 TI - Brain morphometric comparison of first-episode schizophrenia and temporal lobe epilepsy. AB - BACKGROUND: Converging evidence has suggested that the abnormalities in brain morphology observed in schizophrenia are similar to those seen in temporal lobe epilepsy (TLE). The purpose of this study was to compare the features of these groups directly with measures of the brain using magnetic resonance (MR) morphometry. METHOD: Morphometric measures of ventricular and hippocampal volumes obtained from FLASH MR images were studied in 32 patients with first-episode schizophrenia (FES), 39 patients with TLE (21 left, 18 right), and 42 healthy controls. RESULTS: Ventricular volumes in the FES and TLE groups were both significantly larger that those seen in controls and did not differ from each other. The FES group showed significantly larger temporal horns, while the TLE group had relatively larger frontal horns. Analyses of hippocampal volumes revealed a significant group by hemisphere effect. The FES group showed relative reductions in left hippocampal volume that were comparable only to TLE patients with seizures originating from the left hemisphere. CONCLUSION: The results indicate that FES and TLE groups both show evidence of ventricular enlargement. Lateralised morphological abnormalities of the hippocampal formation in FES and left TLE are comparable, and may be specific to temporolimbic regions. PMID- 9330017 TI - Predictive power and construct validity of the Level of Expressed Emotion (LEE) scale. Depressed out-patients and couples from the general community. AB - BACKGROUND: The Level of Expressed Emotion scale (LEE) is a questionnaire designed to measure the perception of expressed emotion, an important predictor of the course of several psychiatric disorders. METHOD: In this study, the scale's predictive and construct validity were examined in a sample of 26 clinically depressed out-patients and their partners, and in a sample of 40 couples from the general community. RESULTS: In the sample of depressed out patients, the LEE was predictive of depression improvement at six-month follow up. With regard to the construct validity, results in both samples showed quite strong relationships between the LEE and depressive symptomatology, relational dissatisfaction, and coping styles. CONCLUSIONS: The LEE may be a useful tool in the study of interpersonal processes and depression, both in clinical and research settings. PMID- 9330018 TI - No association between bipolar disorder and alleles at a functional polymorphism in the COMT gene. Biomed European Bipolar Collaborative Group. AB - BACKGROUND: There is compelling evidence for the existence of susceptibility genes for bipolar disorder. Association studies using functional DNA variations are an important approach for identifying these genes. The enzyme catechol-O methyltransferase (COMT) plays a key role in the degradation of catecholamine neurotransmitters and is a candidate for involvement in bipolar disorder. Recently a common functional genetic polymorphism that underlies population variation in COMT activity has been elucidated and a simple assay developed. METHOD: In a collaboration involving seven European centres, we have undertaken an association study of this functional polymorphism in 412 unrelated West European caucasian DSM-III-R bipolar patients and 368 ethnically matched controls. RESULTS: We found no evidence of allelic or genotypic association. CONCLUSIONS: We can conclude that variation at this functional polymorphism does not make an important contribution to bipolar disorder in the Western European population. Future studies using this powerful experimental approach can be expected to contribute to identification of bipolar susceptibility genes. PMID- 9330019 TI - Mental health in primary care. An epidemiological study of morbidity and use of health resources. AB - BACKGROUND: The main objectives of the survey were: (a) to analyse the sociological, clinical and illness-related correlates of mental illness in primary care; (b) to study, during one-year follow-up, outcome and use of health resources. METHOD: The survey comprised a two-phase cross-sectional study. In the first phase patients were classified using the GHQ-28 or by the general practitioner (GP). In the second phase they were assessed by the SCAN system. RESULTS: The prevalence rate of mental illness (in attenders) using the GHQ was 33.2%. The corresponding rate for the GP was 14.1%, and for the SCAN 31.5%. Mental illness mainly comprised depression, anxiety and alcohol-related diagnoses. The presence of mental illness and the use of health resources during follow-up were dependent on demographic characteristics and on their original psychiatric status. CONCLUSIONS: In primary care, mental illness constitutes a major health problem. Despite this fact, GPs do not recognise a substantial proportion of these health problems. PMID- 9330020 TI - The effect of physical ill health on the course of psychiatric disorder in general practice. AB - BACKGROUND: The aim of this study was to determine the effect of physical morbidity on the outcome of patients with psychiatric disorder, and to compare the effects on non-medically and medically explained symptoms. METHOD: One hundred and fifty psychiatric cases were recruited using a two-stage design from 1620 consecutive patients attending their GP. Subjects were assessed at the time of screening, and one year subsequently, using the Composite International Diagnostic Instrument adapted for use in primary care (CIDI-PHC) and the Groningen Social Disability Schedule (GSDS). Assessments of psychiatric morbidity were also obtained from GPs. RESULTS: Medically explained somatic symptoms were strongly related to psychological outcome one year later. Whereas just over a half of patients with no medically explained symptoms had recovered from a psychiatric disorder, the percentage recovery fell to 41% in those with 1-4 medically explained symptoms, and only 21% in patients with five or more medically explained symptoms. CONCLUSIONS: Physical ill-health has been shown to make an independent contribution to psychological outcome. The specific needs of these patients should receive greater attention. PMID- 9330021 TI - The familial aggregation of common psychiatric and substance use disorders in the National Comorbidity Survey: a family history study. AB - BACKGROUND: Most family studies of psychiatric disorders examine one syndrome at a time, and identify probands in clinical rather than epidemiological settings. METHOD: In the National Comorbidity Survey, 5877 respondents were asked about the history of five psychiatric disorders in their parents: major depression (MD), generalised anxiety disorder (GAD), antisocial personality disorder (ASP), alcohol abuse/dependence (AAD) and drug abuse/dependence (DAD). RESULTS: Significant familial aggregation was seen for all disorders. Controlling for other disorders produced only modest reductions in the odds ratios for MD, GAD and AAD and larger reductions for ASP and DAD. The familial transmission of these disorders can be explained by underlying vulnerabilities to internalising and to externalising disorders transmitted across generations with moderate fidelity. CONCLUSIONS: Familial aggregation of common psychiatric and substance use disorders is substantial in epidemiologic samples. The examined environmental adversities account for little of the observed parent-offspring transmission of these conditions. PMID- 9330022 TI - The effect of citalopram in panic disorder. AB - BACKGROUND: Citalopram is a serotonin reuptake inhibitor which has been demonstrated to be highly selective and with a superior tolerability profile to the classical tricyclic antidepressants. This study was designed to test whether there was any difference in efficacy in the management of panic disorder (PD) between citalopram and placebo. METHOD: This was a double-blind, placebo and clomipramine controlled, parallel group eight-week study. A total of 475 patients with PD, with or without agoraphobia, were randomised to treatment with either placebo, clomipramine 60 or 90 mg/day, or citalopram 10 or 15 mg/day, or 20 or 30 mg/day, or 40 or 60 mg/day. Doses were increased over the first three weeks, stabilised during the fourth week and fixed between weeks five and eight. RESULTS: Treatment with citalopram at 20 or 30 mg, 40 or 60 mg and clomipramine were significantly superior to placebo, judged by the number of patients free of panic attacks in the week prior to the final assessment. All rating scales examined suggested that citalopram 20 or 30 mg was more effective than citalopram 40 or 60 mg. CONCLUSION: The most advantageous benefit/risk ratio for the treatment of PD was associated with citalopram 20 or 30 mg/day. PMID- 9330024 TI - Predicting PTSD in trauma survivors: prospective evaluation of self-report and clinician-administered instruments. AB - BACKGROUND: This study examined the ability of commonly used questionnaires and a structured clinical interview to predict PTSD in recent trauma survivors. METHOD: Horowitz's Impact of Event Scale (IES), Speilberger's State Anxiety (SANX) and the Peri Traumatic Dissociation Questionnaire (PDEQ) were administered one week post-trauma to 239 traumatised individuals recruited from a general hospital emergency room. The IES, the SANX, the civilian version of the Mississippi Scale for Combat Related PTSD (MISS), and the Clinician Administered PTSD Scale (CAPS) were administered one month and four months post-trauma. Receiver operator characteristic (ROC) analysis was used with these data. RESULTS: All questionnaires were better than chance at predicting PTSD. The so-called PTSD questionnaires (IES and MISS) were not better than the more general ones. No difference in predictive value was found when questionnaires were carried out one week or one month after a trauma. Recovery was better predicted than PTSD, and the CAPS was better than the questionnaires. DISCUSSION: The use of psychometrics and clinical interviews to predict PTSD should be guided by clinical relevance and by the availability of resources. PMID- 9330023 TI - Prolactin response to d-fenfluramine in obsessive-compulsive patients, and outcome of fluvoxamine treatment. AB - BACKGROUND: Although several studies have directly explored serotonin (5-HT) transmission in patients with obsessive-compulsive disorder (OCD), their results have been inconsistent and their clinical relevance is doubtful. METHOD: According to a double-blind placebo-controlled design, plasma prolactin (PRL) response to a specific serotonergic probe, d-fenfluramine, was measured in 20 drug-free obsessive-compulsive patients and in 20 matched healthy controls. After the neuroendocrine test, 15 patients completed a 10-week treatment with fluvoxamine. Psychopathological assessment was performed before and after therapy. RESULTS: PRL response in OCD patients was blunted under the drug-free condition; correlated inversely with pretreatment ratings of obsessive-compulsive and depressive symptomatology; and correlated inversely with the improvement in obsessive-compulsive score observed after fluvoxamine treatment. CONCLUSIONS: These results support the idea of a dysfunction of 5-HT transmission in OCD, and suggest that the greater this impairment, the better the response to drugs which selectively block the reuptake of 5-HT. PMID- 9330025 TI - Confusional State Evaluation (CSE): an instrument for measuring severity of delirium in the elderly. AB - BACKGROUND: Delirium or confusional state is a common mental disorder in the elderly. To follow changes in symptoms over time and to evaluate the efficacy of treatment of delirium, a reliable and valid instrument for measuring degrees of delirium is essential. METHOD: An observer's rating scale with 22 items, the Confusional State Evaluation (CSE), was developed. Scores on 12 of the items were summarised to a "confusion score". Based on ratings of 71 demented and non demented elderly patients with delirium, the interrater reliability and validity of the scale was studied. RESULTS: Agreement between two independent raters was fair to excellent (weighted kappa 0.38-0.93). The correlation between the "confusion score" of the scale and the global rating by a psychogeriatrician was good (r = 0.79). CONCLUSIONS: The CSE seems to be a reliable and valid measuring instrument which can be useful in following the course of confusion in elderly patients. PMID- 9330026 TI - Clinical specificity of prison inmates with severe mental disorders. A case control study. AB - BACKGROUND: We wished to determine whether prison inmates with severe mental disorders possess specific clinical characteristics compared with psychiatric in patients suffering from similar problems. METHOD: Under a case-control design, 69 male prison inmates suffering from a schizophrenic or major affective disorder were matched for age and diagnostic spectrum to 60 male psychiatric in-patients. Standardised interviews were used to diagnose psychiatric disorders according to DSM-III-R and social functioning criteria. Case-notes were reviewed to cull data regarding social life, criminal record and service use. RESULTS: Inmates were more likely to suffer from delusional/NOS psychotic disorders (72%) or major depression (70%), and psychiatric in-patients from schizophrenic or bipolar disorder (62% and 71%, respectively). Comorbidity was more prevalent among inmates than among in-patients, while in-patients presented less social autonomy than did inmates. CONCLUSIONS: The clinical specificity of prison inmates with severe mental disorders clearly differentiates them from psychiatric in-patients, and warrants recognition of their special needs for assessment and integrated treatment approaches. PMID- 9330027 TI - Influenza and schizophrenia. PMID- 9330028 TI - Early detection of schizophrenia. PMID- 9330030 TI - Terminology of learning disability. PMID- 9330029 TI - Terminology of learning disability. PMID- 9330031 TI - Confidential inquiry into suicide and homicide by mentally ill people. PMID- 9330032 TI - Measuring cognitive deterioration in Alzheimer's disease. PMID- 9330033 TI - CYP2D6 genotype and tardive dyskinesia. PMID- 9330034 TI - Dissociative pathology discriminates between bipolar mood disorder and dissociative disorder. PMID- 9330035 TI - Simultaneous spectrophotometric determination of calcium and magnesium in mineral waters by means of multivariate partial least-squares regression. AB - A method for simultaneous spectrophotometric determination of calcium and magnesium in mineral waters using multivariate calibration methods is proposed. The method is based on the development of the reaction between the analytes and Methylthymol Blue at pH 11. Two operational modes were used: static (spectral information) and flow injection (FI) (spectral and kinetic information). The selection of variables was studied. A series of synthetic solutions containing different concentrations of calcium and magnesium were used to check the prediction ability of the partial least-squares models. The method was applied to the analysis of mineral waters and the results were compared with those obtained by complexometry. No significant differences at the 95% confidence level were found. The proposed method is simple, accurate and reproducible, and it could be easily adapted as a portable (static mode) or automatic (FI) method. PMID- 9330036 TI - Probability for detecting hot particles in environmental samples by sample splitting. AB - The presence of radioactive hot particles in environmental samples (e.g., soil, vegetation, sediments) is frequently detected by observing significant differences in the activities of sub-samples, which are otherwise alike. The probabilities for detecting hot particles in this way were calculated by using Monte Carlo methods as a function of the number of hot particles in the original sample, the number of sub-samples used, the frequency distribution of the activities of the hot particles, and the precision with which the activities of the sub-samples are determined. Assuming, for example, (i) a log-normal distribution of the activities of the hot particles with a relative standard deviation eta > or = 1, and (ii) that a difference of > 30% between the activities of the sub-sample with the largest and that with the smallest activity can be detected, splitting the original sample into three sub-samples will be sufficient to detect the presence of up to five hot particles with a probability of > 95%. If four sub-samples are used, the presence of up to 20 hot particles can be detected with this probability. In general, it will not be effective to increase the precision of the activity measurements of the sub-samples at the expense of the number of sub-samples investigated. PMID- 9330037 TI - Derivatization of amphetamine and methamphetamine with 1,2-naphthoquinone 4 sulfonic acid into solid-phase extraction cartridges. Determination of amphetamine in pharmaceutical and urine samples. AB - The derivatization of amphetamine and methamphetamine with 1,2-naphthoquinone-4 sulfonate (NQS) into solid-phase extraction cartridges (C18) is described. Optimum conditions were the use of carbonate-hydrogencarbonate buffer of pH 10, a 10 min reaction time at 25 degrees C and an NQS concentration of 9.6 x 10(-3) M. The accuracy and the precision of the method were tested. The results obtained with the proposed liquid-solid procedure were compared with those obtained with a traditional liquid-liquid extraction with hexane-ethyl acetate. The procedure was used to measure amphetamine in pharmaceutical and urine samples. PMID- 9330038 TI - Fully automatic on-line separation preconcentration system for electrothermal atomic absorption spectrometry: determination of cadmium and lead in sea-water. AB - An automatic separation preconcentration system coupled to an electrothermal (graphite furnace) atomic absorption spectrometer is described. The preconcentration step is performed on a chelating resin microcolumn (Chelex-100) placed in the injection tip of the autosampler. A time based manifold with two- and three-way solenoid valves commanded by an eight channel microcomputer programmable controller is used for column conditioning, preconcentration and washing steps; no manual operations are involved. Elution is performed by the programmable graphite furnace autosampler and achieved in only one step. Operations involving complete and partial injection of the eluate into the graphite furnace are also discussed. The system was applied to the determination of Cd and Pb in near shore sea-water from Patagonia, Argentina. Detection limits of 1 and 8 ng l-1 were obtained for Cd and Pb respectively. Analysis of a certified reference material (CASS-3) showed good agreement with the certified values. PMID- 9330039 TI - Chemiluminescence flow system for vanadium(v) with immobilized reagents. AB - A chemiluminescence (CL)-based system for vanadium(v) combined with flow injection analysis is described. The analytical regents, luminol and hexacyanoferrate(II), were both immobilized on an anion-exchange resin column. When a volume of phosphoric acid was passed through the column, these two reagents were eluted from the resin and then mixed with a vanadium(v) stream under acidic conditions. By means of the fast oxidation reaction between vanadium(v) and hexacyanoferrate(II), vanadium(IV) and hexacyanoferrate (III) were generated, both of which catalyzed the oxidation of luminol by dissolved oxygen in aqueous alkaline solution to produce CL. The CL emission intensity was correlated with the standard vanadium (v) concentration in the range from 1.0 x 10(-2) to 10 micrograms cm-3, and the detection limit was 5.4 x 10(-3) micrograms cm-3 vanadium (v). Interfering metal ions co-existing in sample solutions could be effectively separated on-line by a cation-exchange column placed upstream. A complete analysis, including sampling and washing, could be performed in 1 min with a relative standard deviation of less than 5%. The system was stable for over 100 analyses and was applied successfully to the determination of vanadium in geochemical and human hair samples. PMID- 9330040 TI - Determination of norfloxacin in real samples by different spectrofluorimetric techniques. AB - Simple, rapid, accurate and sensitive spectrofluorimetric methods for the determination of norfloxacin are described. The methods are based on the reaction of this drug with aluminium(III) ion to form a strongly fluorescent complex. Fluorescence properties of the AlIII-norfloxacin complex were used for the determination of this drug in pharmaceutical preparations. First-derivative constant wavelength synchronous fluorescence spectrometry was used for the determination of norfloxacin in the presence of nalidixic acid. The determination of norfloxacin in urine without the need of tedious pre-separation was achieved by using zero-crossing second-derivative synchronous fluorescence spectrometry. PMID- 9330041 TI - Determination of lactic acid and poly(lactic acid)s in a dermatological formulation by capillary electrophoresis. AB - A stability-indicating capillary electrophoresis (CE) method was developed to determine lactic acid at a level of 0.9% m/m in a dermatological formulation. This level includes up to 34% present in the form of poly(lactic acid)s. Current methods which involve treatment of samples with sodium hydroxide to hydrolyse the various lactic acid oligomers to free lactic acid cannot strictly be regarded as true measures of stability. The harsh hydrolysis conditions used can also lead to the generation of many degradation peaks from excipients such as natural extracts which can interfere with the detection of lactic acid in HPLC and CE methods. Application of indirect UV detection using 4-methoxybenzoic acid (p-anisate) as the background electrolyte and negative voltage polarity (detector towards anode) allows the direct quantification of the total available lactic acid from the monomer and the predominant linear oligomers in the raw material, i.e., the dimer (lactoyllactate) and the trimer. Tetradecyltrimethylammonium bromide was used as the electroosmotic flow modifier, lactic acid, lithium salt, as a calibration standard and butyric acid as an internal standard. A typical RSD for standard response using migration time-corrected peak area ratios was 1.17% (n = 9). Duplicate analysis of total lactic acid from two similar product placebos spiked with raw material gave average recoveries of 101.0 and 99.6%. The method, when applied to fully and partially hydrolysed raw material, showed good agreement (99.6%) with a standard titration assay for raw materials and was successfully applied to 5 month storage samples of a product. Advantages of the method include speed, simplicity, low consumption of reagents and no organic solvents. PMID- 9330043 TI - Dandruff and seborrhoeic dermatitis: causes and management. PMID- 9330044 TI - A left-right comparison of UVB phototherapy and topical photochemotherapy in bilateral chronic hand dermatitis after 6 weeks' treatment. AB - We have compared the efficacy of local UVB phototherapy with topical (bath) photochemotherapy in 13 patients with bilateral chronic hand dermatitis. In each patient, one hand was treated with UVB phototherapy and the other hand with topical (bath) photochemotherapy. Both treatments moderately improved the chronic hand dermatitis after 6 weeks' treatment. We observed no significant differences in improvement between the modalities, but side-effects occurred more often on the photochemotherapy-treated side. Considering the similar responses and relative incidence of side-effects, we would advise starting treatment with UVB phototherapy and only using topical photochemotherapy if this fails. PMID- 9330045 TI - The relationship of Ki67 and involucrin expression in proliferative, pre neoplastic and neoplastic skin. AB - In normal skin, proliferation and differentiation are tightly coupled in order to maintain normal architecture in a continually renewing tissue. The temporal and spatial relationships between these two processes in normal, psoriatic, pre neoplastic and neoplastic skin were investigated by a double immunolabelling technique with Ki67 as a marker of proliferation and involucrin as a marker of terminal differentiation. In normal skin, expression of the two antigens was strictly spatially segregated. In the abnormal, the proportions of cells expressing the antigens were increased with some loss of the spatial segregation, while small numbers of cells showed dual expression suggesting loss of the normal control between proliferation and differentiation. However, the quantitative ratio of proliferation to differentiation in psoriatic and pre-neoplastic skin was similar to the normal; transition to an invasive phenotype, however, was associated with a reversal of this ratio, and this correlated well with the degree of histological differentiation. PMID- 9330042 TI - Determination of disodium cromoglycate in human urine by high-performance liquid chromatography with post-column photoirradiation-fluorescence detection. AB - For the determination of disodium cromoglycate in urine, a fluorimetric method using HPLC post-column photoirradiation has been developed. The mobile phase consisted of a 35 mmol l-1 phosphate buffer (pH 8)-methanol (7 + 3, %v/v) containing 75 mmol l-1 hydrogen peroxide and 20 mmol l-1 18-crown-6. The 18-crown 6 was used for separation adjustment of the disodium cromoglycate in the urine sample. Photoirradiation was carried out in tubing wound around a germicidal light in a reactor equipped with an air-cooling fan. The fluorescence was monitored with excitation at 325 nm and emission at 448 nm. The calibration graph for disodium cromoglycate was linear over the range 38-2340 ng ml-1 using an injection volume of 100 microliters. The pretreatment of the urine samples consisted of diluting and filtering steps. The mean recovery of disodium cromoglycate from urine was 99.1 +/- 2.4% (n = 6). PMID- 9330047 TI - A novel method for estimating the volume of capillary haemangioma to determine response to treatment. PMID- 9330046 TI - Retrospective study of the epidemiology of nodular vasculitis followed up in the National Skin Centre, Singapore. AB - A seven-year retrospective study was done on histologically proven nodular vasculitis presenting at the National Skin Centre, Singapore. We collected 25 patients and found that they differed from the classical European model. They most commonly involved the shins and did not ulcerate. A tuberculous aetiology accounted for 28% of cases. Useful indicators of tuberculosis were a past history of tuberculosis, an abnormal chest X-ray and a strongly positive Mantoux test. PMID- 9330048 TI - Verrucous carcinoma in association with hypertrophic lichen planus. AB - Neoplastic transformation of lichen planus is a rare event. However, squamous cell carcinoma may develop in 0.3%-3% of patients with the oral form of the disease. On the other hand, less than 30 cases arising in cutaneous lichen planus have been reported, and only four cases of verrucous carcinoma in association with either form, one with an oral lesion and three with cutaneous lesions (one hypertrophic and one ulcerative). This report describes the unusual progression of a hypertrophic lichen planus plaque of the right leg to a verrucous carcinoma in a 40-year-old woman. PMID- 9330049 TI - Acne fulminans and erythema nodosum during isotretinoin therapy responding to dapsone. AB - Acne vulgaris is very common, 85% of teenagers being affected at any one time. In most cases, the disease is mild and patients do not present to the dermatologist. Most are instead treated with over-the-counter products and conventional treatment such as peeling agents or topical and systemic antibiotics. Isotretinoin has revolutionized the treatment of severe acne unresponsive to oral antibiotics. Explosive and very severe acne such as pyoderma faciale, acne conglobata and acne fulminans are rare, the features that distinguish acne fulminans from the other conditions being systemic upset with fever, joint pain, malaise and leucocytosis, while there have been two reports of the condition associated with erythema nodosum. The recommended treatment for acne fulminans is a combination of oral steroids and systemic antibiotics, isotretinoin probably not being the treatment of choice. We now report a patient who developed acne fulminans and erythema nodosum within 3 weeks of starting isotretinoin and then responded to dapsone without oral steroids. PMID- 9330050 TI - Nylon cloth macular amyloidosis. AB - In primary localized cutaneous amyloid, deposition of amyloid is confined to the skin without the involvement of any internal organs. Amyloid deposition in the skin is often scanty, and electron microscopy may be needed to confirm the presence of the typical amyloid fibrils. There have been several case reports of cutaneous amyloidosis associated with friction or rubbing of the skin. We report a case of primary localized cutaneous amyloid associated with the habitual use of a nylon cloth. PMID- 9330051 TI - Bullous pemphigoid evolving into cicatricial pemphigoid? AB - We describe three patients who initially presented with both clinical and immunological findings to support a diagnosis of bullous pemphigoid but whose subsequent course has been that of cicatricial pemphigoid. Mucosal scarring was accompanied by a fall in autoantibody titres in our three patients. These cases illustrate the difficulties clinicians may experience in assigning a specific diagnosis to patients. They also support the concept that bullous pemphigoid and cicatricial pemphigoid are part of a single disease spectrum. The most intriguing question is what specific factors determine the expression of a particular disease phenotype as bullous pemphigoid and cicatricial pemphigoid share target antigens and also the DQ7 allele. PMID- 9330052 TI - Dowling Degos disease in association with multiple seborrhoeic warts. AB - A patient is described with histopathological features in keeping with a diagnosis of Dowling Degos disease but with some unusual clinical features. Other members of the family are similarly affected. PMID- 9330053 TI - Transient leukaemia cutis in chronic lymphocytic leukaemia. AB - Leukaemia cutis arises due to cutaneous infiltration of neoplastic leukocytes or their precursors. Recent evidence suggests that this sign does not necessarily herald a poor prognosis. We describe a 72-year-old woman with B-cell chronic lymphatic leukaemia (CLL) who developed a papular eruption on her breast at the site of a recent herpetic eruption. Histology and immunostaining showed a dense dermal B-cell infiltrate in keeping with leukaemia cutis. The papules cleared in 6 months without treatment, leaving atrophic scars. The histological features and possible aetiological mechanisms of post-herpetic papular eruptions in CLL are reviewed. PMID- 9330054 TI - Kaposi's varicelliform eruption in a patient with mycosis fungoides. AB - A 39-year-old Japanese woman suffering from plaque stage of mycosis fungoides (MF) developed Kaposi's varicelliform eruption (KVE) on her face. KVE appeared 1 month after commencing skin electron beam irradiation (SEBI), when the total irradiation had reached 46 Gy. Natural killer (NK) cell activity in the peripheral blood was abnormally low, but returned to normal after 1 year. However, lymphocyte reactivity to mitogens was persistently low. These facts suggested that the KVE in our patient developed because of abnormally low NK cell reactivity probably induced by radiation therapy. PMID- 9330055 TI - Pellagra, azathioprine and inflammatory bowel disease. PMID- 9330056 TI - Postirradiation angiosarcoma. AB - We describe a patient with angiosarcoma of the scalp arising in an area of radiodermatitis caused by X-ray epilation therapy for scalp ringworm as a child. Radiotherapy has been well documented as a causative factor in the formation of cutaneous malignancies, most notably basal cell and squamous cell carcinoma, but only rarely angiosarcoma. This is now the first reported case of angiosarcoma arising after X-ray epilation therapy for scalp ringworm, and we postulate a causative link between the two events. PMID- 9330057 TI - PUVA for vitiligo and skin cancer. PMID- 9330058 TI - Concurrent Sweet's syndrome (acute febrile neutrophilic dermatosis), erythema nodosum and sarcoidosis. PMID- 9330059 TI - Increased risk of skin cancer in patients with ectodermal dysplasia--a contraindication to psoralen and UVA (PUVA) therapy? PMID- 9330060 TI - Bowen's disease--an unusual case with a revealing past history. PMID- 9330061 TI - The treatment of rosaceous lymphoedema. PMID- 9330062 TI - Pityriasis versicolor associated with oral lithium therapy. PMID- 9330063 TI - Chronic actinic dermatitis associated with primary biliary cirrhosis. PMID- 9330064 TI - Serum levels of soluble interleukin-2 receptor--a possible index of disease prognosis in systemic sclerosis. PMID- 9330065 TI - Linear lichen planus and lichen striatus: is there an intermediate form between these conditions? PMID- 9330066 TI - Acrokeratoelastoidosis: a report of two sporadic cases. PMID- 9330067 TI - Volume-weighted mean nuclear volume of basal cell carcinoma and risk of recurrence. PMID- 9330068 TI - Systemic treatment of malignant melanoma: grade is as important as stage. AB - The prognosis of malignant melanoma relates to the biological profile of the tumour and is independent of the magnitude of surgery on the offending primary lesion. The optimal management of this cancer throughout its evolution depends on the appraisal of its metastatic potential. With better understanding of the natural history of the disease and improvements in the therapeutic ratio of currently available drugs, systemic treatment of high risk melanoma in the adjuvant context and before dissemination is emerging as a therapeutic priority. There is now evidence from recently published, controlled studies with long follow up that this approach can prolong relapse-free and overall survival. PMID- 9330069 TI - Comparison of serum antibiotic levels in acne patients receiving the standard or a modified release formulation of minocycline hydrochloride. AB - Serum levels of minocycline hydrochloride were determined by bioassay in a total of 223 acne patients (123 male, 100 female) receiving either the recommended dose (100mg/day) or a high dose (200mg/day) of the standard preparation (101 patients) of a modified release formulation (132 patients). Sera were collected within 6 h of the morning dose 7-10 days after the start of treatment. Mean minocycline serum levels were consistently higher in females than in males, irrespective of dose or formulation. The differences only reached statistical significance (P < 0.05, Student's t-test) in the case of the standard preparation at a dose of 50 mg, b.d. Serum levels were increased significantly in both sexes at the higher dosage of each formulation (P < 0.01) but there was no significant difference between formulations at either dosage. Variation in serum concentrations was not accounted for by variation in body mass. Serum levels above the modal minimum inhibitory concentration (MIC) of minocycline for fully sensitive strains of Propionibacterium acnes I (0.125 micrograms/mL) were recorded in all patients. In contrast, serum levels equal to or greater than the modal MIC of minocycline for resistant propionibacteria (2 micrograms/mL) were recorded in only 17.9% of patients on the low dose standard preparation compared with 55.6% on the high dose standard preparation (P < 0.001, chi 2). Even in females on the high-dose modified release formulation, 32.2% had serum levels below the modal MIC of minocycline for resistant strains. We conclude that, in terms of achievable serum levels over a short time period, there is no advantage of the modified release formulation over the standard preparation of minocycline. Whichever formulation is used, dose manipulation may be necessary to achieve maximum therapeutic benefit, especially in those individuals who are colonized by propionibacteria with reduced sensitivity to minocycline. PMID- 9330070 TI - The value of direct immunofluorescence as a diagnostic aid in dermatomyositis--a study of 35 cases. AB - Dermatomyositis (DM) in a inflammatory disorder of skeletal muscle and skin closely related to other connective tissue diseases; however, to date, no conclusive immunofluorescence (IMF) data are available for the disorder. The aim of this study was therefore to analyse retrospectively the clinical, histological and direct IMF findings for a group of 35 patients who had presented during an 8 year period. Clinically, 29 of the patients had the typical cutaneous features of DM, while 16 had evidence of myositis based on electromyographic or muscle biopsy findings; six developed malignant tumours during the study period, three of which proved fatal. Cutaneous histopathology findings were compatible with the diagnosis of DM in 18 cases, with evidence of a lupus band in six. Direct IMF showed a lupus band at the basement membrane zone in 19 (with IgM, IgG or C3), with colloid bodies in seven. Serologically, only nine were antinuclear antibody positive, one extractable nuclear antibody positive and all 35 anti-Jo-1 negative. Eight had other circulating autoantibodies, namely thyroid (three), gastric parietal cell (two), smooth muscle (two) and rheumatoid factor (one). Our findings suggest that direct IMF can be a useful adjunct in the investigation of patients with DM, and may prove helpful in diagnosis when both the histopathology and serology are inconclusive. PMID- 9330071 TI - IgE bullous disease. AB - We report two patients with the typical picture of bullous pemphigoid who lacked two critical diagnostic immunopathological features of the disease, namely IgG or C3 bound to the epidermal basement membrane and circulating IgG antibodies directed against the basement membrane zone (BMZ). Both patients had dense infiltrates of eosinophils within their skin lesions, as well as markedly elevated serum IgE levels, while immunofluorescent studies with anti-IgE antibody revealed heavy IgE deposition on inflammatory cells within the dermis surrounding the bullae. These cells were confirmed to be eosinophils by means of specific staining with antibody to major basic protein (MBP). We speculate that this 'IgE bullous disease' resulted from IgE-mediated hypersensitivity induced by focal infection, both patients initially being helped by antibiotics. However, dramatic clearing of bullae was seen following surgical removal of a battery implant (Patient 1), and bilateral above-the-knee amputations of gangrenous legs (Patient 2). PMID- 9330072 TI - The impact of onychomycosis on quality of life. AB - Onychomycosis is a common disease, which is estimated to affect approximately 3% of the UK population. It is often viewed as being relatively trivial in nature; however, there has been little research to evaluate how this condition affects patients' physical and psychological well-being. The aim of this study was to assess the possible effect of onychomycosis on different aspects of patients' quality of life. PMID- 9330073 TI - Linear IgA bullous dermatosis of childhood: treatment with dapsone and co trimoxazole. PMID- 9330074 TI - Pyoderma gangrenosum of the breast treated with low-dose cyclosporin A. AB - Pyoderma gangrenosum (PG) is a painful chronic ulcerative skin disorder often occurring in association with systemic disease. It typically affects the lower limbs, but may also involve other sites, or sometimes develop after trauma of surgical procedures. We report the case of a woman with rheumatoid arthritis who developed disfiguring and severe PG of the right breast, a rare site, following biopsy for a benign breast lesion, and who was subsequently successfully treated with low-dose cyclosporin A. PMID- 9330075 TI - Segmental scarring following intrauterine herpes simplex virus infection. AB - We report the case of a female infant with an intrauterine herpes simplex type II infection in zosteriform distribution. She was treated with several courses of intravenous acyclovir leading to healing of the skin with segmental scarring. This patient is unusual in that the infection occurred in zosteriform distribution without any evidence of systemic involvement. PMID- 9330076 TI - A case of acanthosis nigricans in obese siblings with a pedigree of familial polyposis coli. PMID- 9330077 TI - Proliferative myositis: an unusual cause of multiple subcutaneous nodules. AB - Proliferative myositis is a rare inflammatory condition which clinically has an apparently aggressive growth pattern reminiscent of a sarcoma. Histopathological features are characteristic but not widely recognized, so misdiagnosis may occur. We describe a patient presenting to the Dermatology Department and review the literature and current diagnostic techniques. PMID- 9330078 TI - Angiocentric T cell lymphoma of the skin presenting as inflammatory nodules of the leg. AB - We describe two cases of malignant lymphoma presenting as inflammatory nodules of the leg and mimicking panniculitis clinically. In both cases the skin biopsies showed prominent involvement of the subcutaneous tissue by lymphoma cells. In addition, lymphoma cells invaded blood vessels in the dermis or the subcutaneous tissue. One case was characterized by predominantly extravascular and intravascular location of the lymphoma cells, and the other case by the predominantly extravascular and intramural location of lymphoma cells. These histological findings were compatible with those of angiocentric T-cell lymphoma but with some unusual features. Angiocentric T-cell lymphoma of the skin should be listed in the group of diseases which present as inflammatory nodules of the leg. PMID- 9330079 TI - Nail growth measurement by nail indentation. PMID- 9330080 TI - Progressive granulomatous resection to Bacille Calmette-Guerin (BCG) vaccination. PMID- 9330081 TI - Schleromyxoedema with prominent linear eruption and polyclonal gammopathy. PMID- 9330082 TI - Building a framework for multiple improvement initiatives. AB - BACKGROUND: As health care organizations struggle to compete and even survive in today's turbulent marketplace, they often juggle a variety of initiatives addressing cost reduction, quality improvement, critical pathways, accreditation, clinical guidelines, strategic planning, and organizational development-each with its own priorities, advocates, methods, and language. EXAMPLE: One large health care system that had adopted continuous quality improvement (CQI) as a major strategy then responded to significant cost pressures with additional initiatives in reengineering, cost reduction, and physician guidelines, with varying connections to the quality language, principles, and methods. The senior leadership committee for the quality initiative undertook the development of a framework to explain the connections among the diverse projects and approaches-a framework still in use after two years. SUMMARY AND CONCLUSION: In this period of enormous change, health care leaders must quickly and effectively mobilize all available resources to optimize organizational effectiveness. Too often, management's response patterns have overemphasized a "splitting" tactic, managing multiple distinct initiatives. The creation of integrated delivery systems, mergers, acquisitions, and alliance exacerbates this problem, as the number of initiatives multiplies in the new organizations. Development and application of an integrating management framework will accelerate the pace of organizational improvement by increasing shared understanding of the current desired states of the organization; linking strategic, cultural, and method needs and behaviors; aligning various management initiatives in relation to organizational goals and one another; increasing the fit of management methods with specific situations; and providing a unifying perspective and language for organizational members to act and learn collaboratively. PMID- 9330083 TI - Evaluating and improving pressure ulcer care: the VA experience with administrative data. AB - BACKGROUND: A number of state initiatives are using databases originally developed for nursing home reimbursements to assess the quality of care. Since 1991 the Department of Veterans Affairs (VA; Washington, DC) has been using a long term care administrative database to calculate facility-specific rates of pressure ulcer development. This information is disseminated to all 140 long term care facilities as part of a quality assessment and improvement program. DATA ON PRESSURE ULCER DEVELOPMENT: Assessments are performed on all long term care residents on April 1 and October 1, as well as at the time of admission or transfer to a long term care unit. Approximately 18,000 long term care residents are evaluated in each six-month period; the VA rate of pressure ulcer development is approximately 3.5%. Reports of the rates of pressure ulcer development are then disseminated to all facilities, generally within two months of the assessment date. IMPLICATIONS FOR OTHER QUALITY IMPROVEMENT EFFORTS: The VA's more than five years' experience in using administrative data to assess outcomes for long term care highlights several important issues that should be considered when using outcome measures based on administrative data. These include the importance of carefully selecting the outcome measure, the need to consider the structure of the database, the role of case-mix adjustment, strategies for reporting rates to small facilities, and methods for information dissemination. CONCLUSION: Attention to these issues will help ensure that results from administrative databases lead to improvements in the quality of care. PMID- 9330084 TI - Assessing influenza immunization rates in Medicare managed care plans: a comparison of three methods. AB - BACKGROUND: In an effort to improve care delivered to Medicare beneficiaries, the Health Care Financing Administration (HCFA) has encouraged competitive Medicare risk plans to collaborate on quality improvement projects. PRO-West, a private, nonprofit quality improvement organization, fostered a collaboration of all Medicare risk plans in Washington State in order to assess and improve influenza immunization rates among seniors enrolled in managed care. METHODOLOGY: After the 1994-1995 influenza immunization season, immunization rates were determined for each participating plan from administrative data and medical record review. In the 1995-1996 season, these methods were supplemented with a telephone survey. The survey was used to identify perceived barriers to immunization and to estimate immunization rates. RESULTS: Immunization rates, as estimated by administrative data and medical record review, were similar for both years. The average immunization rate using administrative data for the 1995-1996 flu season was 60.6% (range, 42.8% to 72.3%). The estimated rate increased to 77.8% (95% confidence interval, 75.3% to 80.3%) when the telephone survey data were added. Medical record review contributed little additional information. CONCLUSIONS: Influenza immunization rates for seniors enrolled in Medicare risk plans in Washington State exceed those reported for fee-for-service Medicare beneficiaries. Telephone surveys resulted in higher and probably more valid estimates of influenza immunization rates than did analysis of administrative data and medical records. Plans with lower rates can emulate "benchmark" plans that are explicit about the methods they use to achieve high coverage rates. Medicare risk health plans competing in the same markets can collaborate in quality assessment activities in an effective manner. PMID- 9330085 TI - Providing patients the information they need. PMID- 9330123 TI - The mysterious angiotensin-converting enzyme inhibitors. PMID- 9330124 TI - Clinical and neurohumoral differences between spirapril and captopril in mild to moderate chronic congestive heart failure. AB - BACKGROUND: This study was done to determine whether the difference in duration of action of the long-acting angiotensin-converting enzyme (ACE) inhibitor spirapril compared with the short-acting ACE inhibitor captopril affects clinical efficacy in patients with congestive heart failure. METHODS AND RESULTS: The effects on exercise capacity, neurohumoral status, and quality of life were studied in 20 patients with mild to moderate congestive heart failure in a double blind, randomized, comparative study in parallel groups with a duration of 12 weeks. All assessments during the study were performed in the morning, before intake of the study medication, to avoid the expected peak effect of the ACE inhibitors used. Mean peak oxygen consumption (peak Vo2) was 17.4 mL/min/kg (range, 14.2-19.9 mL/min/kg) and mean left ventricular ejection fraction was 28% (range, 13-40%). Exercise duration in the captopril group showed a significant increase after 12 weeks (P < .05) of treatment compared with the spirapril group. Peak oxygen consumption tended only to increase in the captopril-treated patients compared with the spirapril-treated patients. Serum ACE activity was significantly different between the two treatment groups during treatment (P < .0001) and showed only a significant decrease in the spirapril group. There was no difference in improvement of quality of life between the two treatment groups. CONCLUSIONS: This study showed that the effects of the ACE inhibitors spirapril and captopril on exercise capacity are not related to the degree of inhibition of serum ACE activity. PMID- 9330125 TI - Safety and efficacy of carvedilol in severe heart failure. The U.S. Carvedilol Heart Failure Study Group. AB - BACKGROUND: Many patients remain markedly symptomatic despite optimal current therapy for heart failure. Beta-blockers have often been viewed as contraindicated in this group because of their potential adverse short-term effects on cardiac function. METHODS AND RESULTS: One hundred thirty-one patients with severe congestive heart failure were enrolled into a double-blind, placebo controlled study of the vasodilating beta-blocker carvedilol. All patients had symptomatic, advanced heart failure while on standard triple therapy, as evidenced by a mean ejection fraction of 0.22, marked reduction in distance traveled in a 6-minute corridor walk test, and severe impairment in quality of life measured by the Minnesota Living With Heart Failure Questionnaire. After a 2 week, open-label test of 6.25 mg twice daily carvedilol, 105 patients were randomized (2:1) to receive either carvedilol (up to 25 mg twice daily, n = 70) or matching placebo (n = 35) for 6 months while background therapy with digoxin, diuretics, and an angiotensin-converting enzyme inhibitor remained constant. Ten patients (8%) did not complete the open-label period because of adverse events and 11.4% in both the carvedilol and placebo groups dropped out in the double blind phase. The study was terminated early by the Data Safety and Monitoring Board and follow-up evaluation was therefore aborted before the projected number of patients and follow-up time was achieved. Quality of life, which was the primary endpoint, improved similarly in the carvedilol and placebo groups, whereas the global assessment by the physicians and the patient exhibited a better response to carvedilol (P < .05). Hospitalization and mortality rate were too low to evaluate a difference, and exercise time and New York Heart Association classification did not change significantly in response to the drug. Left ventricular ejection fraction rose significantly (+0.09) in the carvedilol group compared with the placebo group (+0.02, P = .004). CONCLUSION: The beta blocker carvedilol can be safely employed in patients with severe heart failure. Improved left ventricular function with a trend for some improvement in symptoms combined with the experience with the drug in the larger population of less severe patients in this multicenter trial suggests that carvedilol may have a favorable long-term effect in heart failure of diverse severity. PMID- 9330126 TI - Respiratory oxygen cost for dead space challenge is characteristically increased during exercise in patients with chronic heart failure: does it further decrease exercise capacity? AB - BACKGROUND: Although the work of the respiratory muscles is markedly increased during exercise in patients with chronic heart failure, the role of this abnormality in exercise intolerance is still controversial. This issue may be clarified directly by dead space challenge, as this technique increases minute ventilation. Therefore, in this study, the effects of an external dead space on exercise ventilation, gas exchange data, and exercise capacity in patients with chronic heart failure were examined. METHODS AND RESULTS: Dead space challenge was performed by adding an external dead space to the airway in 20 patients with chronic heart failure and 10 normal subjects. Two hours after completion of the control maximal bicycle exercise, the second exercise was performed under application of an external dead space equivalent to 10% of peak tidal volume. Respiratory gas exchange data were collected during exercise. Aerobic exercise capacity was assessed from the exercise time and the time to anaerobic threshold. The sensation of exertional dyspnea was assessed using Borg's rating scale. As compared with data during the control exercise, minute ventilation was increased by approximately 25% with the external dead space throughout exercise in both groups. A parallel 20% increase in systemic oxygen uptake was observed in the heart failure group, likely reflecting an increase in respiratory muscle work. This response was not observed in the normal group. Despite an additional increase in respiratory muscle work, neither aerobic exercise capacity nor exertional dyspnea was exacerbated in the heart failure group by the external dead space. CONCLUSIONS: Dead space challenge appears to be a unique technique that characteristically increases the work of respiratory muscles during exercise in patients with chronic heart failure. By use of this technique, it was demonstrated that an increase in respiratory muscle work is not important in reducing exercise capacity of patients with chronic heart failure. PMID- 9330127 TI - Effects of captopril on interstitial collagen in the myocardium after infarction in rats. AB - BACKGROUND: Myocardial infarction is an important cause of heart failure because it cause tissue loss and contractility disturbances. In chronically infarcted hearts the increase in the collagen content in the extracellular matrix of the surviving viable myocardium has been considered a major factor contributing to development of heart failure. Postinfarction neuroendocrine activation involving the renin-angiotensin system has been implicated in this cardiac fibrosis. METHODS AND RESULTS: As collagen synthesis and degradation are dynamic processes and postinfarction remodeling is a time-dependent phenomenon, rats submitted to coronary artery ligation to produce myocardial infarction were treated with captopril after infarction (30 mg/kg, intraperitoneally, daily) to investigate whether blockade of the renin-angiotensin system can prevent postinfarction myocardial hypertrophy and reactive fibrosis. Groups of rats with myocardial infarction were treated with captopril throughout the protocol period (6 weeks), or during the first 3 weeks after infarction (early therapy), or only during the last 3 weeks of the protocol (late therapy). Untreated groups of rats with or without myocardial infarction were used as control subjects. All animals were killed 6 weeks after surgery to evaluate hypertrophy of heart chambers and collagen deposition in the right ventricle wall and in surviving left ventricular muscle. Protein and hydroxyproline concentrations were assayed biochemically in these tissue homogenates. Only rats with an infarct covering 20 to 40% of the left ventricular surface were included in the study. In the control uninfarcted group (n = 12), hydroxyproline content was 152 +/- 12 micrograms in the right ventricle and 370 +/- 30 micrograms in the left ventricle. These values increased (P < .05) to 232 +/- 13 and 630 +/- 46 micrograms, respectively, in the group with myocardial infarction (n = 8) without treatment. These values were significantly reduced (P < .05) to 160 +/- 9 micrograms in the right ventricle and 520 +/- 40 micrograms in the left ventricle in the group with myocardial infarction treated with captopril for 6 weeks. The percentage decreases in collagen content and myocardial weight produced by captopril were similar. Thus, hydroxyproline concentration (mg hydroxyproline muscle), which increases significantly in both ventricles after myocardial infarction, was not modified by captopril therapy. Protein concentration in the right and left ventricular muscles decreased after myocardial infarction. This decrease was enhanced in the infarcted groups submitted to captopril treatment, mainly in the group treated for 6 weeks. Lesser effects on hypertrophy and hydroxyproline content were observed in the groups of rats treated with captopril in only the earlier or later phase of infarction. CONCLUSIONS: It is concluded that captopril reduces similarly postinfarction hypertrophy and collagen deposition in surviving myocardium. These effects, although less intense, also occur when the drug is used for a short period immediately after myocardial infarction or when used later, when ventricular remodeling is almost fully developed. PMID- 9330128 TI - Effects of selective dopaminergic receptor stimulation on ventricular remodeling after experimental myocardial infarction in rats. AB - BACKGROUND: The disappointing results of the nonselective dopaminergic agonist ibopamine in the treatment of heart failure may be caused by nonselective receptor stimulation. Therefore, a search for more selective dopaminergic agonists remains important. Z1046 is such a compound. METHODS AND RESULTS: Forty two normotensive rats with a myocardial infarction (MI) and 18 sham-operated rats were studied. Rats with MI were treated for 6 weeks with Z1046 (n = 12) or ibopamine (n = 12) or were not treated (n = 18). Sham-operated control rats were not treated (n = 18). Assessments during the trial included those of plasma catecholamine levels, cardiac function, and morphology. Z1046 significantly decreased heart rate and blood pressure in rats with MI. Ibopamine affected only blood pressure. Compared with control rats with MI (736 +/- 66 pg/mL), plasma norepinephrine was significantly lower both after Z1046 (508 +/- 44 pg/mL) and after ibopamine (561 +/- 28 pg/mL). Infarct size was significantly reduced both by Z1046 and by ibopamine (P < .05). Z1046, but not ibopamine, normalized baseline left ventricular pressure (P < .05, treated rats vs MI control rats). CONCLUSIONS: The new (relatively selective) dopaminergic agonist Z1046 appears to have a more pronounced effect in protection against remodeling than ibopamine; this results in preservation of cardiac function. The effects appear to be mediated by both a reduced sympathetic drive and an improved hemodynamic profile. PMID- 9330129 TI - Myocardial reactive hyperemia in experimental chronic heart failure: evidence for the role of K+ adenosine triphosphate-dependent channels and cyclooxygenase activity. AB - BACKGROUND: Several studies suggest that coronary perfusion is abnormal in heart failure. The fact that these deficits may results in an altered coronary reserve remains controversial. Therefore, coronary adaptability to short-duration ischemia and the resultant myocardial reactive hyperemia were investigated in a model of chronic heart failure. METHODS AND RESULTS: Experiments were performed in normal and failing hamster hearts (UM-X7.1, aged > 225 days). Heart rate, left ventricular developed pressure, and coronary flow were recorded continuously before and after each 30-second ischemia in isolated perfused heart preparations. Studies were conducted under control conditions and in the presence of four inhibitors of potential mediators of the reactive hyperemia response: the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (30 microM), the adenosine antagonist 8-(p-sulfophenyl)theophylline (50 microM), the K+ cyclic adenosine triphosphate-dependent channel antagonist glibenclamide (10 microM), and the cyclooxygenase inhibitor indomethacin (10 microM). Baseline hemodynamic parameters were all significantly impaired in failing hearts. Under control conditions, failing hearts were able to respond adequately to a 30-second ischemia: repayment-to-debt ratio averaged 1.02 +/- 0.09 as compared with 1.10 +/ 0.09 in normal hearts (P = NS). All inhibitors significantly reduced basal coronary perfusion except for indomethacin. Of the four inhibitors of potential mediators of the myocardial reactive hyperemic response, only glibenclamide and indomethacin impaired the repayment-to-debt ratio. In their presence, repayment to-debt ratio was reduced by 40% of the baseline response (P < .01) without significant difference between normal and failing hearts. On the contrary, NG nitro-L-arginine methyl ester and 8-(p-sulfophenyl)theophylline did not alter the repayment-to-debt ratio. CONCLUSIONS: These observations demonstrate the capacity of the failing heart to tolerate short-duration ischemia despite the presence of significant alterations in its basal coronary perfusion. In addition, results suggest that activation of K+ adenosine triphosphate-dependent channels and the presence of cyclooxygenase by-products are important determinants of coronary adaptation to short-duration ischemia in this model of chronic heart failure. PMID- 9330131 TI - Diastolic dysfunction in heart failure. PMID- 9330130 TI - The propionyl-L-carnitine hypothesis: an alternative approach to treating heart failure. AB - Propionyl-L-carnitine (PLC) is a naturally occurring compound that has been considered for the treatment of congestive heart failure (CHF). The rationale for its use in this pathology is related to its effects on cardiac and skeletal muscle. Chronic treatment with PLC improves the contraction of isolated and aerobic perfused rabbit hearts. The compound improves energy metabolism and myocardial contractility in different experimental models of heart failure, such as pressure-overloaded rats, infarct model of heart failure, and rabbit with streptozotocin-induced diabetes. In general, the effect of PLC is apparent in situations of high energy demand such as those induced by increased workload. It therefore seems likely that PLC is able to correct some metabolic steps of the process that leads to heart failure. In addition, PLC may be helpful in heart failure because of its specific action on peripheral skeletal muscle. Administration of PLC in patients with CHF improves skeletal muscle metabolism by increasing pyruvate flux into the Krebs cycle and by decreasing lactate production. These effects occur in the absence of major hemodynamic and neuroendocrinologic changes and may underlie the ability of PLC to increase exercise performance in patients with heart failure. In a randomized study of 50 patients with mild CHF, PLC increased the maximal exercise time, reduced lactate production, and improved left ventricular ejection fraction. There have been two large-scale trials on the effects of PLC on both cardiac and peripheral muscle function in CHF. One is ongoing; the other one, which just ended, failed to show an improvement in exercise capacity in the population studied. A benefit was evident only in a subgroup of patients with preserved ejection fraction and impaired baseline exercise duration. PMID- 9330132 TI - Arthroscopic diagnosis of intra-articular soft tissue injuries associated with distal radial fractures. AB - Arthroscopy was used to assess the soft tissue injuries associated with distal radial fractures in 118 acute intra- and extra-articular fractures. The triangular fibrocartilage complex (TFCC) was torn in 46 of 118 patients--in 35% of intra-articular fractures and in 53% of extra-articular fractures. No correlation between ulnar styloid fractures and TFCC injuries could be found. Scapholunate (SL) ligament injuries with instability were present in 21.5% of intra-articular fractures and in 6.7% of extra-articular fractures. Lunotriquetral (LT) ligament injuries with instability were present in 6.7% of intra-articular fractures. and in 13.3% of extra-articular fractures. Combined SL and LT injuries were present in 5.6% of intra-articular fracture. Preoperative radiographs correlated with TFCC injury. Patients with TFCC tears had greater shortening and dorsal angulation on the preoperative radiographs. Preoperative radiographs had no predictive value for interosseous ligament injury. Ligamentous injuries are commonly associated with both intra-articular and extra-articular distal radial fractures. PMID- 9330133 TI - Prospective multicenter trial of a plate for dorsal fixation of distal radius fractures. AB - A new plate designed specifically to address complex wrist pathology was used for the internal fixation of 22 complex fractures of the distal radius in 22 patients in a prospective multicenter trial. The majority of fractures were group C2- and C3-type fractures according to the Comprehensive Classification of Fractures. No plate failures, loss of reduction, nonunions, or infections occurred. Within the average follow-up time of 14 months, the functional results (including an average motion of 76% and an average grip strength of 56% of the contralateral side) were comparable to those reported for similar fractures in previous investigations. Five patients had irritation of the tendons in the second dorsal compartment. This trial serves both as a verification of the safety and efficacy of this distal radius plate as well as a demonstration of its utility in the treatment of complex fractures of the distal radius. PMID- 9330134 TI - Opening-wedge osteotomy, bone graft, and external fixation for correction of radius malunion. AB - A technique of radius opening-wedge osteotomy, bone graft, and external fixation for the treatment of symptomatic radius malunion is presented. It provides direct rigid fixation to the osteotomy components, thus maintaining the correction while allowing early wrist exercises. This technique has been effective for 7 patients in correcting deformities that averaged--20 degrees palmar tilt with radial shortening of 3.4 mm to a postoperative average palmar tilt of 5.3 degrees and 0.4 mm radial shortening. It is an alternative technique for the hand surgeon treating radius malunion and can be easily combined with adjunctive procedures. PMID- 9330135 TI - Radiographic evaluation of osseous displacement following intra-articular fractures of the distal radius: reliability of plain radiography versus computed tomography. AB - This study evaluated the reliability of plain radiography versus computed tomography (CT) for the measurement of small (< 5 mm) intra-articular displacements of distal radius fracture fragments. The plain radiographs and CT scans of 19 acute intra-articular distal radius fractures were used by 5 independent observers, using 2 standardized techniques, to quantify incongruity of the articular surface in a blinded and randomized fashion. Repeat measurements were performed by the same observers 2-4 weeks later, allowing determination of intraclass correlation coefficients (ICC) as a measure of intraobserver and interobserver agreement. The average maximum gap displacement on plain radiographs was 2.1 mm (range, 0.0-15.0 mm, lateral view) and on CT images was 4.9 mm (range, 0.7-17.3 mm, axial view). The average maximum step displacement on plain radiographs was 0.9 mm (range, 0.0-6.4 mm, lateral view) and on CT images was 1.2 mm (range, 0.0-6.0 mm, sagittal view). More reproducible values determining step and gap displacement were obtained when the arc method of measurement was used on CT scans (ICC values, .69-.97) as compared to the longitudinal axis method for plain radiographs (ICC values, .30-.50). For measured displacements of 2 mm or more, our data demonstrated poor correlation between measurements made on CT images and those made on plain radiographs (gap or step displacement > 2 mm, K = 0.21; step displacement > 2 mm, K = 0.21). Thirty percent of measurements from plain radiographs significantly underestimated or overestimated displacement compared to CT scan measurements. From these data, we conclude that CT scanning data, using the arc method of measurement, are more reliable for quantifying articular surface incongruities of the distal radius than are plain radiography measurements. PMID- 9330136 TI - Kinematics of the scaphoid shift test. AB - Twenty-five uninjured subjects (50 wrists) were examined clinically and fluoroscopically during performance of the scaphoid shift test. Wrists were placed into 3 groups on the basis of the degree of palpable carpal motion that occurred during the clinical examination. Kinematic parameters of rotation and displacement were calculated from digitized images of the carpals at rest and at maximum displacement. On clinical exam, 36% of normal individuals had positive findings on scaphoid shift test. Dorsal displacement of the scaphoid was not significantly associated with positive scaphoid shift test results in these subjects, while total displacement of the scaphoid (the sum of axial and dorsal displacement) was significantly associated with positive test results. The principle confounding factor appeared to be a high degree of displacement that occurred at the capitolunate joint in some individuals, termed a "midcarpal shift." The data demonstrate that despite a high prevalence of positive scaphoid shifts among uninjured individuals, the ability to accurately detect dorsal displacement of the scaphoid using the scaphoid shift test is limited. On the basis of their findings, the authors recommend that positive test results be confirmed fluoroscopically. PMID- 9330137 TI - Ulnar shortening combined with arthroscopic repairs in the delayed management of triangular fibrocartilage complex tears. AB - The functional outcome after surgery was determined in 21 patients an average of 29 months (range, 24-52 months) after surgery to evaluate the efficacy of arthroscopic repair of triangular fibrocartilage complex (TFCC) tears and ulnar shortening. All of the patients had reparable lesions of the TFCC treated after a delay of more than 6 months from the time of injury. The patients' average age was 32 years and all patients had wrist pain limiting them from work and/or sports prior to surgery. After surgery, there was a significant relief of pain (p < .01). Grip strength and range of motion averaged 83% +/- 18% and 81% +/- 16%, respectively, of that of the uninjured side. At follow-up evaluation, 14 patients with repairs underwent follow-up studies; the TFCC was noted to be intact in 12 patients. PMID- 9330138 TI - Operative technique for inside-out repair of the triangular fibrocartilage complex. AB - A technique for arthroscopic inside-out repair of peripheral traumatic (type 1B) tears of the triangular fibrocartilage complex is reported. The technique can be performed using zone-specific cannulas that are commonly used for repairing meniscal tears in the knee. Anatomic dissections were used to show the safe regions around the TFCC where tears are amenable to this type of repair. PMID- 9330139 TI - Tendon repair--cellular activities in rabbit deep flexor tendons and surrounding synovial sheaths and the effects of hyaluronan: an experimental study in vivo and in vitro. AB - One deep flexor tendon and its surrounding sheath of each hindpaw of 48 rabbits were transected and repaired in order to investigate the abilities of rabbit flexor tendons and synovial sheaths to synthesize DNA and matrix components during healing and to study the effects of hyaluronan (HA). After repair, HA or saline was injected between the tendon and the sheath. Short-term culture and labeling in vitro were used up to 6 weeks after surgery to determine synthesis of DNA, proteoglycan, collagen, and noncollagen protein. Within tendon repair sites, the rate of cell proliferation increased and reached a maximum 5 days after surgery; within repaired synovial sheaths, the rate immediately decreased. In the healing tendons, the rate of collagen synthesis decreased and the rate of noncollagen protein synthesis remained unchanged. The opposite results were found within the healing synovial sheaths. HA did not affect the rate of cell proliferation or matrix synthesis in healing tendons or surrounding sheaths. These results show that cellular activities differ between tendons and synovial sheaths during healing and that those activities may not be affected by HA. PMID- 9330140 TI - Reduction of restrictive adhesions by local aprotinin application and primary sheath repair in surgically traumatized flexor tendons of the rabbit. AB - The effects of microsurgical and medical treatments on reduction of adhesions in surgically traumatized flexor tendons of rabbits are quantified in this study. The effects of the mentioned techniques were investigated for the following 4 groups: (1) neither primary sheath repair nor aprotinin application was done, (2) primary sheath repair was done but no aprotinin was used, (3) primary sheath repair was not done but local aprotinin (15,000 IU/kg) was applied, and (4) primary sheath repair was done and local aprotinin was applied. At the sixth and twelfth postoperative weeks, the flexor digitorum profundus tendons of the second and the third digits were subjected to biomechanical tests. Only the third digit was used in macroscopic and histopathologic evaluations. There were 6 digits included in each subgroup of biomechanical tests and 4 digits per subgroups in macroscopic and histopathologic evaluations. Work of flexion (WOF) values were obtained by calculating the area under the load-displacement curve. Percent resistive work of flexion (PRWOF) was obtained by calculating the difference between the WOF value for the repaired right digit and the WOF value for the contralateral corresponding nonrepaired digit. Combined primary sheath repair and medical treatment yielded the best results in reducing the restrictive adhesions in injured tendons. The differences between the PRWOF values of group 4 were 33.7% +/- 8.2% and 15.8% +/- 7.7% for the sixth and twelfth postoperative weeks, respectively. The corresponding values for group 1 were 95.7% +/- 13.8% and 51.75% +/- 10.25%. PMID- 9330141 TI - Staged extensor tendon reconstruction in the finger. AB - Staged extensor tendon reconstruction using a silicone implant followed by tendon grafting was done to restore proximal interphalangeal (PIP) joint extension in 6 fingers with severe injuries to the dorsal skin and extensor mechanism. Abrasions to the joint capsule and cortical surfaces were also present. To avoid finger stiffness, the reconstruction was delayed and range of motion exercises were initiated early. The skin injury was managed by split-thickness skin grafting or allowed to heal by secondary intention to avoid prolonged immobilization. During surgery, the peritendinous fascia of the extensor tendon is used to guide insertion of the implant, and it serves as a premade tunnel that appears to aid the gliding and stability of the implant and subsequent tendon graft. Active extension of the PIP joint was restored in all fingers; there was an average extension lag of 15 degrees. PIP joint flexion averaged 95 degrees. On the basis of this experience, the author believes the technique to be a reliable treatment alternative for severely injured fingers with extensor mechanism loss. PMID- 9330142 TI - Biomechanical analysis of four-strand extensor tendon repair techniques. AB - Experience with flexor tendon repairs has suggested the superiority of the augmented Becker (MGH) technique for strength, toughness, and gap resistance. In an effort to apply these findings to the extensor tendons, 3 four-strand extensor tendon repair techniques were biomechanically tested in fresh human cadaver limbs: modified Bunnell, modified Krackow-Thomas, and MGH. Repairs were performed in Verdan's zone VI. Repaired tendons were distracted at constant speed until rupture. Tendon load and tendon distraction were continuously monitored. Benchmark values for load were measured as fingers were pulled from full metacarpophalangeal (MP) joint flexion to full extension, to 1-mm gap formation at the tenorrhaphy, and to complete rupture of the repair. The MGH repair proved significantly more resistant to gap formation (stronger and tougher) than the Bunnell and Krackow-Thomas repairs (p < .02). No differences were seen between groups in repair performance at MP joint extension and at complete rupture. This study suggests that the MGH technique has superior gap resistance to the other four-strand methods tested for extensor tendon repair in Verdan's zone VI. The MGH repair is recommended for extensor tendon repairs in zone VI when early postoperative motion regimens are considered. PMID- 9330143 TI - Measurement of thumb abduction strength: normative data and a comparison with grip and pinch strength. AB - Abduction strength of the thumb was measured in normal men (n = 101; age range, 21-94 years) and women (n = 208; age range, 20-97 years). Abduction-strength measurements were conducted concurrently with grip and pinch strength measurements made by well-established clinical methods. Normal values were established and stratified by age and sex. Thumb abduction strength generally correlated with grip and pinch strength. All strength variables at all ages were greater in men than in women. The magnitude of all strength variables was maintained from 20 to 59 years of age, then decreased with increasing age in both men and women. Measurement of thumb abduction strength may prove to be a useful adjunct to the various tests currently used by hand surgeons to assess hand function. PMID- 9330144 TI - The effects of low median nerve block on thumb abduction strength. AB - The relative contributions of the abductor pollicis longus (APL) and abductor pollicis brevis (APB) to thumb abduction strength (TAS) were determined after a selective nerve block in 21 normal volunteers. The median nerve was anesthetized (blocked) at the wrist. Needle electromyography verified paralysis of the APB and usually the opponens pollicis; in 6 study subjects, the superficial head of the flexor pollicis brevis (FPB) was also paralyzed. The APL, innervated by a branch of the posterior interosseous nerve, remained functionally intact. TAS was measured by a mechanical device before and after median nerve block. Median nerve block at the wrist resulted in a dramatic decrease in TAS in all volunteers. The mean loss of TAS was 70.3% in men and 74.3% in women. Postblock TAS tended to be greater in those subjects with retained function of the FPB. This study verifies that TAS is primarily a function of the APB. PMID- 9330145 TI - Ultrasonographically assisted carpal tunnel release. AB - An operative technique of carpal tunnel release using intraoperative ultrasonography is described. In this technique, "safe line" is defined in the transverse carpal ligament and the adjacent deep forearm fascia midway between the ulnar margin of the median nerve and the radial margin of the ulnar artery. After ultrasonographic design of a 1.0 to 1.5-cm skin incision along the safe line at the distal carpal tunnel, the distal ligament is released under direct vision. Proximal release is performed along this line under ultrasonographic monitoring using a device that consists of a basket punch and an outer metal tube. In a prospective randomized study, the outcomes were compared for carpal tunnel release using either this technique in 50 hands of 50 patients or conventional open release in 53 hands of 53 patients. Follow-up assessment at 3, 6, 13, 26, 52, and 104 weeks showed no significant difference with respect to numbness and paresthesias, static two-point discrimination, findings on Semmes Weinstein monofilament testing, findings on manual muscle testing of the abductor pollicis brevis, and electrophysiologic findings. The ultrasonographic-release group had better outcomes regarding pain, tenderness of the scar, and key-pinch strength at 3, 6, and 13 weeks, and grip strength at 3 and 6 weeks after surgery. The scar was more aesthetic in this group. There were no complications with either technique. PMID- 9330146 TI - Cubital tunnel release and medial epicondylectomy: effect of timing of mobilization. AB - The effects of early versus late range of motion (ROM) following cubital tunnel decompression and medial epicondylectomy were evaluated in a randomized prospective study. Forty-five consecutive procedures were divided into 2 groups. The early mobilization group started rehabilitation at an average of 3 days after surgery and the late mobilization group started rehabilitation at an average of 14 days after surgery. Flexion contracture of more than 5 degrees degrees was observed in 5% of the early mobilization group, compared to 52% of the late mobilization group (p < .001). On average, patients in the early mobilization group returned to work twice as early as those in the late mobilization group and did not experience any adverse effects on their grip strength or other hand functions. Institution of ROM exercises immediately after surgery was found to be more effective in preventing elbow flexion contractures, obtaining a quicker recovery, and allowing return to work with no ill effects. PMID- 9330147 TI - Evaluation of cutaneous vibration thresholds in medical transcriptionists. AB - This study was designed to determine whether vibration thresholds of transcriptionists varied significantly from the thresholds of individuals not exposed to keyboard activities. Using a multifrequency vibrometer, we obtained vibration threshold values from 31 medical transcriptionists who perform work on computer keyboards and compared them to values obtained from 40 control subjects. Thresholds tended to become more abnormal at higher frequencies, although this difference was statistically significant only at frequencies of 125 Hz, 250 Hz, and 500 Hz in the index and small fingers. Vibration thresholds were not found to increase significantly with age or years of occupation. Vibration thresholds were significantly increased in medical transcriptionists at the higher frequencies, suggesting subtle neural dysfunction. PMID- 9330148 TI - Association between a quantitative measure of tactile acuity and hand symptoms reported by operators of power tools. AB - An association between a quantitative measure of tactile acuity at the fingertips and symptoms of reduced manipulative function, as established by responses to a questionnaire, was demonstrated in a population of 81 manual workers from the mining industry (62 power-tool operators and 19 nonusers). Mechanoreceptor specific vibrotactile thresholds were determined for the slowly adapting type I (SAI) and fast-adapting types I and II (FAI and FAII) receptors at the fingertip of the third digit of each hand. Statistically significant threshold shifts in SAI and/or FAII acuity were found in persons responding affirmatively to questions concerning finger/hand numbness, blanching, and difficulty buttoning clothing. The best predictors of a quantitative change in tactile acuity were questions relating to difficulty manipulating small objects and buttoning clothing, yielding positive predictive values of from 90% to 100% and false positive rates of from 0% to 2.8%. The demonstration of an association between a quantitative measure of tactile acuity at the fingertips and some symptom reports, obtained by means of a questionnaire, provides the basis for the development of a screening procedure for persons at risk of such disturbances in hand function. PMID- 9330149 TI - Use of outcome instruments to compare workers' compensation and non-workers' compensation carpal tunnel syndrome. AB - Validated outcome instruments were used to compare treatment outcomes of carpal tunnel syndrome (CTS) in workers' compensation and non-workers' compensation patients. A self-administered questionnaire consisting of the generic Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the disease-specific Carpal Tunnel Syndrome Instrument was mailed to 277 patients randomly selected from all 1050 new patients treated for CTS during a 1-year period. A total of 212 patients (61 workers' compensation and 151 non-workers' compensation) responded to the survey 7-22 (mean, 14) months after the initiation of treatment, yielding a response rate of 76%. Workers' compensation patients had worse mean scores than non-workers' compensation patients in 6 of the 8 SF-36 scales and in the 2 Carpal Tunnel Syndrome Instrument scales, but validating multivariate analysis could not verify significant score differences in any of the scales. Thus, this study could not demonstrate inferior treatment outcomes of CTS in workers' compensation patients as measured by standardized generic and disease-specific outcome instruments. PMID- 9330150 TI - Radial tunnel syndrome: long-term results of surgical decompression. AB - Between 1980 and 1990, 31 patients (33 extremities) underwent decompression of the radial tunnel. All procedures were performed at the same institution by the senior author using a brachioradialis muscle-splitting approach. Twenty-three patients (24 extremities) were available for follow-up evaluation at an average of 8 years after surgery. The outcome was determined using the original criteria of Roles and Maudsley and Ritts et al. By the criteria of Roles and Maudsley, outcomes for 15 patients (16 extremities; 67%) were rated as excellent/good and for 8 patients (8 extremities; 33%), as fair/poor. By the criteria of Ritts et al., 16 patients (17 extremities; 71%) had good results and 7 patients (7 extremities; 29%), a fair/poor result. Overall, the majority of patients were satisfied and felt helped by the surgery. Five patients changed their occupation because of continued discomfort. Significant differences in outcome were not found for workers' compensation patients. Similarly, no differences in outcome were found for site of compression, patient sex, history of trauma, or associated neuropathies. The results of this study suggest that complete pain relief and return to activities following radial tunnel surgery is not as predictable as previous studies have indicated. Nineteen patients (20 extremities), however, felt satisfied and helped and believed they gained improved function because of surgical decompression of the radial tunnel. PMID- 9330151 TI - Comparison of transthecal and subcutaneous single-injection digital block techniques in cadaver hands. AB - A controlled, randomized single-blinded study was performed on the thumbs and little fingers of 20 cadaver hands. The digits were randomly divided into 2 groups. In the transthecal group, 2 mL of 0.5% methylene blue was injected into the tendon sheath at the A1 pulley. In the subcutaneous group, the same amount of dye was injected into the subcutaneous tissue superficial to the A1 pulley. The injections were performed by 2 investigators. They exchanged specimens and performed dissections on the injected digits without knowledge of which technique had been used. The distributions of dye along the digit and the color intensity of the dye on the digital nerves were studied. There was no significant difference (p > .05) between results for the 2 techniques. It was expected that both techniques would result in similar anesthetic distribution in the clinical setting. In the transthecal group, intra-articular staining of the metacarpophalangeal joint was noted in 3 little fingers and 1 thumb. This complication did not occur in the subcutaneous group. This difference was significant (p < .05). PMID- 9330152 TI - Comparison of transthecal and subcutaneous single-injection digital block techniques. AB - A randomized double-blinded study was performed on 20 normal volunteers to evaluate 2 different techniques of single-injection digital anesthesia. Single injection transthecal digital block technique was used to anesthetize 1 index finger and single-injection subcutaneous technique to block the other index finger. Pain and light touch were evaluated and sensory nerve-conduction studies were performed on both index fingers. These data were obtained prior to the nerve blocks and then at 10-minute intervals until recovery from the anesthesia. The method of anesthesia was found to have no effect on the distribution, onset, and duration of anesthesia. Median and radial nerve sensory nerve action potential amplitude reductions following digital anesthesia were also not influenced by the technique of anesthesia. Single-injection subcutaneous block was found to be easier to administer and to produce less pain during and 24 hours after injection than did the single-injection transthecal technique. PMID- 9330153 TI - Continuous cervical epidural anesthesia in reconstructive hand surgery. AB - Continuous cervical epidural anesthesia was used for 17 operations in 16 patients undergoing immediate reconstructive surgery after upper-extremity injuries or tumor resection and was continued for postoperative pain management. Routine hemodynamics, arterial blood gases, plasma bupivacaine levels, and skin temperatures were recorded before and after the block. The surgery time ranged from 3 to 18 hours. Postoperative pain management was maintained for up to 6 days. The blocks were adequate for surgery and postoperative pain treatment in all cases. There were no signs or symptoms indicating local anesthetic toxicity. Circulatory and respiratory integrity was well maintained. The patients were all ambulatory the day after surgery and could start physiotherapy immediately. This regional anesthesia technique may have significant advantages over branchial plexus block or general anesthesia for lengthy surgical procedures of the upper extremity. PMID- 9330154 TI - Resurfacing of the donor defect after wrap-around toe transfer with a free lateral forearm flap. AB - In thumb reconstruction, the wrap-around free flap has many advantages; however, delayed wound healing, pain, and skin ulcerations at the donor site can be a problem. In 5 patients, a free lateral forearm fasciocutaneous flap was successfully used for immediate resurfacing of the donor defect of the big toe during wrap-around procedures. This flap was selected after preliminary anatomic studies that showed that it could be safely raised if based on the anterior terminal division of the posterior radial collateral artery. The average follow up period was 2 years. The time required for healing of the great-toe defect was less than 1 month. All patients were satisfied with the outcome of the procedure. The skin of this flap is very pliable and thin, and the subcutaneous tissue is about half the thickness of that of the lateral arm flap. This technique is especially indicated for closure of moderate to big skin defects at the great-toe level, whenever a larger than usual amount of skin is required, during wrap around procedures for thumb reconstruction. PMID- 9330155 TI - Pisotriquetral loose bodies. AB - Eight patients--2 men and 6 women (mean age, 49 years)--who underwent excision of pisotriquetral (PT) loose bodies were identified from clinic records. The time interval from onset of symptoms to surgery averaged 18 months. Four patients reported a traumatic onset of symptoms, and 4 reported an insidious onset. For all patients, treatment by nonsteroidal anti-inflammatory drugs, splinting, and steroid injection had failed. Routine radiography revealed a loose body in only 4 patients. Trispiral tomography delineated all loose bodies. Three patients underwent loose-body excision only; 5 had PT joint degeneration and underwent additional pisiformectomy. Length of follow-up monitoring averaged 7.4 years. All patients had resolution of wrist pain and improvement in strength. There were no complications. Loose bodies, which may form in the PT joint or migrate from the radiocarpal joint, were identified best by tomography, with simple excision providing excellent relief of symptoms in the absence of PT joint degeneration. PMID- 9330156 TI - Anti-ICAM-1 antibodies protect allografts against microvascular and parenchymal cell damage. AB - Anti-ICAM-1 (anti-intercellular adhesion molecule-1) monoclonal antibodies (CD54) were tested for treating composite tissue allografts in the rat hindlimb cremaster transplantation model for intravital microcirculatory studies. Twenty four transplantations were carried out across major histocompatibility barriers between Lewis-Brown Norway and Lewis rats. Isograft control transplants were compared to nontreated allograft control transplants and to allografts treated with 1 mg/kg of anti-ICAM-1 monoclonal antibodies. At 24 and 72 hours, microcirculatory vessel diameters, red blood cell velocities, functional capillary perfusion, endothelial edema index, and leukocyte-endothelial interactions were measured. At 24 and 72 hours, the number of sticking leukocytes, sticking lymphocytes, transmigrating leukocytes, and the endothelial edema index in the treated allografts were significantly decreased more than in the other 2 groups (p < .05 for all variables). Anti-ICAM monoclonal antibodies significantly reduced leukocyte-endothelial interactions, protecting the allografts from acute microvascular and parenchymal injury. PMID- 9330157 TI - Anatomy of the intermetacarpal ligaments of the carpometacarpal joints of the fingers. AB - In this study, the structure of the retaining ligaments between the proximal metacarpal bones of the fingers was defined. Anatomic dissections were performed on 10 fresh cadavers. Four separate ligaments were found: a dorsal metacarpal ligament, a palmar metacarpal ligament, and 2 interosseous ligaments oriented in a V-shaped configuration. The V-shaped interosseous ligaments were found to be the strongest; along with the palmar and dorsal intermetacarpal ligaments, they form a very strong connection between the bases of the adjacent metacarpals. PMID- 9330158 TI - A cadaver study on volume and surface area of the fingertip. AB - The volume of the volar soft tissue, dorsal soft tissue, and bone and the area of the dorsal and volar surfaces were estimated in 35 adult cadaver fingertips. The fingertip was defined as the part of the finger distal to the plane of the palmar skin crease and the major dorsal crease at the distal interphalangeal articulation. An impression molding technique, involving silicone rubber, was used to determine the volume, while dyed imprints of the dorsal and palmar surfaces were used to determine the surface area. In all digits, the mean volume of the volar soft tissue of the fingertip was found to be about 56%, the dorsal soft tissue about 26%, and the volume of bone about 18%. The volar soft tissue includes the skin and fascia (51% of the total fingertip volume), the flexor tendon and its sheath, and the volar plate and volar joint capsule (5% of the total). Power relationships for the total fingertip volume, the volume of volar soft tissue, and the volume of bone in terms of the length of the fingertip were noted. There was also a linear relationship found between the volume of the volar soft tissue and the volar surface area. This study provides data on the ratio of soft tissue to bone in the fingertip. The maintenance of the soft tissue-to-bone ratio, so as to regain fingertip form and function, may be of particular importance when designing flaps and coverage in the reconstruction of the fingertip. PMID- 9330159 TI - Sarcoidosis of the hand and wrist: a report of two cases. AB - The association of soft tissue and bone sarcoidosis in the hand and wrist is rare. Two cases of sarcoidosis, 1 with involvement of the distal radius and both flexor and extensor tendons in the wrist and another with tenosynovitis of 2 fingers associated with phalangeal osteolysis, are reported. Both patients were surgically treated by tenosynovectomy. The importance of the systemic corticosteroid therapy is emphasized. PMID- 9330160 TI - Clinical perspective: repetitive strain in the workplace. PMID- 9330161 TI - Electrodiagnostic testing in hand surgery. PMID- 9330162 TI - Carpal tunnel pressure. PMID- 9330163 TI - Flexor carpi radialis approach for carpal tunnel release. PMID- 9330164 TI - Risk, diagnosis and management of prosthetic valve endocarditis: a review. AB - Prosthetic valve endocarditis (PVE) emerged approximately 37 years ago when the first human heart valve replacements were performed. PVE can be classified as 'early' or 'late' with the pathophysiology and etiologic organisms varying between the two subgroups. The incidence of PVE ranges up to 0.5% per patient year for mechanical mitral valves and up to 1.0% per patient-year for other valves. The clinical presentation is similar to that of native valve endocarditis, with fever being the most prevalent sign. Diagnosis is based on a constellation of clinical signs and symptoms as well as echocardiographic evaluation of the valve and perivalvular tissues. An algorithm is set forth for diagnosis and management of patients with suspected PVE based on our personal experience and the published literature. Indications for surgery, the surgical approach and methods of PVE prophylaxis and prevention are discussed. PMID- 9330165 TI - The economics of uncomplicated mitral valve surgery. AB - BACKGROUND AND AIMS OF THE STUDY: Comparisons of mitral valve (MV) replacement and reconstruction have demonstrated lower overall complication rates, better left ventricular (LV) function, and inferred overall lower cost for the latter procedure compared with the former. However, assessment of economic differences between the two procedures in routine cases, without complications, has not been reported. This study retrospectively evaluates the economic impact of uncomplicated MV repair versus replacement. METHODS: As this study seeks only to evaluate economic comparisons between routine cases of mitral repair versus replacement, those patients having concomitant procedures performed (coronary revascularization or other valve procedure) or postoperative complications (i.e. pulmonary failure, wound infections, new-onset atrial fibrillation, return for bleeding, or neurologic sequelae) were excluded from the study. Among patients who underwent uncomplicated MV procedures, 30 were selected at random and reviewed. RESULTS: Variables for MV replacement versus reconstruction included aortic cross-clamp time (112 +/- 54 versus 92 +/- 20 min; p = NS), cardiopulmonary bypass (CPB) time (189 +/- 70 versus 128 +/- 18 min; p < 0.05), total hospital stay (8.3 +/- 1.6 versus 5.6 +/- 1.6 days; p < 0.0001), and total hospital charges ($44,697 +/- 4903 versus $31,337 +/- 4484; p < 0.0001), respectively. CONCLUSIONS: These data suggest that, beyond the recognized benefits of MV reconstruction, namely preservation of LV function and avoidance of long-term anticoagulation, there is an economic advantage to MV reconstruction for patients and payors, even in uncomplicated cases. These differences may become more apparent with longer follow-up and in patients having poor function or combined procedures. This finding reinforces the idea that MV reconstruction is the option of choice for patients with mitral regurgitation. PMID- 9330166 TI - Surgical treatment for prolapse of the anterior mitral leaflet. AB - BACKGROUND AND AIMS OF THE STUDY: Mitral valvuloplasty (MVP) for the prolapse of the anterior mitral leaflet (AML) is more difficult than that for the posterior mitral leaflet. The introduction of artificial chordae (November, 1986) and the concomitant maze operation (November, 1992) were surgical 'turning points' in our 17 years' experience. METHODS: In total, 163 surgical cases of AML prolapse based on the above turning points, and carried out between 1979 and 1996, were reviewed. These included 110 MVP and 53 mitral valve replacements. MVP was performed in only 46% (29/63) of patients before October 1986 (Group I); in 72% (42/58) of patients between November 1986 and October 1992 (Group II); and in 93% (39/42) of patients after November 1992 (Group III). RESULTS: Reoperation was required in nine patients. The reoperation-free rate after MVP was 79% at 17 years in all cases, 82% at 17 years in Group I, 86% at 10 years in Group II and 97% at four years in Group III. Besides reoperation cases, grade 3/4 mitral regurgitation (MR), assessed by color Doppler echocardiography, was detected in seven patients. The event-free (reoperation, MR, thromboembolism) rate was 69% at 17 years in all cases, 78% at 17 years in Group I, 71% at 10 years in Group II and 92% at 4 years in Group III. A concomitant maze operation was performed in 19 of 20 current patients with atrial fibrillation. The percentage of sinus rhythm after operation in Group I, II and III was 53%, 60% and 84%, respectively. CONCLUSIONS: During 17 years' experience, mortality and morbidity after MVP for AML prolapse were satisfactory. With the use of artificial chordae, we have been able to perform MVP in more than 90% of current patients with AML prolapse. Further, a concomitant maze procedure could provide a higher incidence of postoperative sinus rhythm. PMID- 9330167 TI - Green Lane Hospital experience with mitral valve repair for prolapse: adverse outcomes for highly symptomatic patients. AB - BACKGROUND AND AIMS OF THE STUDY: Valve repair, where suitable, is the preferred option in patients who require mitral surgery. A number of studies have shown excellent long-term results, but most were undertaken in tertiary referral centers with a high throughput of patients. METHODS: We present our experience in 60 patients, aged 60 +/- 14 years, undergoing repair between 1984 and 1993. Most patients (83%) were in New York Heart Association (NYHA) class II or III at the time of surgery; 27% had concomitant ischemic heart disease. Almost all (98%) had posterior leaflet repair and 18% had anterior leaflet repair. Eight surgeons each performed a mean of 7.5 operations during this period. RESULTS: The 30-day mortality rate was 3.3%. There were seven late deaths. Five patients underwent reoperation for mitral regurgitation (two early, three late). At six years, 60% of patients were alive, or free of stroke or reoperation. Late follow up was obtained in 45 of 47 surviving patients: 95% were in NYHA class I or II; one third were on anticoagulants for atrial fibrillation; 90% had mild (or less) mitral regurgitation on echocardiography. CONCLUSIONS: These data show that most patients have a very good outcome from valve repair surgery and encourage the trend towards operating earlier in the course of the disease. Adverse outcomes occurred mainly in patients who were highly symptomatic at the time of surgery. The high proportion of patients on postoperative anticoagulants underscores the importance of operating before atrial fibrillation becomes permanent. PMID- 9330168 TI - Long-term follow up study on 64 elderly patients after balloon aortic valvuloplasty. AB - BACKGROUND AND AIMS OF THE STUDY: The aims of this study were to evaluate symptomatic improvement and event-free/overall survival after balloon aortic valvulotomy in patients with significant sclerotic aortic valve stenosis. METHODS: Sixty-four patients with calcified aortic stenosis, in NYHA class III IV, and of mean age 79.0 years, underwent a total of 75 scheduled attempts at balloon aortic valvulotomy, with single balloon catheters between December 1987 and June 1993. Patients were either considered as poor surgical candidates or themselves preferred such valvulotomy. RESULTS: Periprocedural major complications, including death in 6%, occurred in association with 16% of the procedures. Among 57 patients in whom initial dilatation was successful, the average period of symptom relief was 9.4 months (median 7.0, range: 0 to 47 months). Independent predictors for longer duration of symptom relief and survival were systolic arterial pressure > 115 mmHg and female gender; ejection fraction > or = 30% was only predictive of survival. Actuarial survival rates at one, two and three years were 77, 48 and 36% respectively. CONCLUSIONS: Balloon aortic valvulotomy is followed by a short period of symptomatic relief and carries a low periprocedural mortality, but considerable morbidity. By comparison, aortic valve replacement patients aged over 70 and with serious physical limitations (NYHA class IIIB-IV) showed much better overall survival. As contraindications to surgery are in most cases relative, aortic valve replacement should always be considered as the only choice in the surgical decision-making. PMID- 9330169 TI - Prediction of stentless aortic bioprosthesis size with transesophageal echocardiography and magnetic resonance imaging. AB - BACKGROUND AND AIMS OF THE STUDY: During stentless bioprosthetic aortic valve replacement, ischemic time may be decreased by the non-invasive prediction of bioprosthesis size, allowing earlier commencement of prosthesis preparation. In this study we examine whether the addition of transesophageal echocardiography (TEE) to transthoracic echocardiography (TTE) aids in the prediction of stentless bioprosthesis aortic valve size. We also report our preliminary experience with the use of magnetic resonance imaging (MRI) in bioprosthetic valve size prediction. METHODS: Eight patients in whom elective aortic valve replacement with a Toronto SPV valve was planned underwent preoperative TTE and MRI, and intraoperative TEE. RESULTS: In all cases the combination of TTE and TEE correctly predicted the size of Toronto SPV valve inserted. In three cases, TEE led to a revision of the TTE-based prediction. The need for sinotubuloplasty in two patients was correctly predicted by both TTE and TEE. MRI of the aortic annulus correctly predicted valve size in three of four cases, but could not reliably identify the sinotubular junction. CONCLUSIONS: In aortic valve replacement the accuracy of prediction of stentless bioprosthesis size is improved by the addition of TEE to TTE. PMID- 9330170 TI - Tanning revisited for leaflets and the aortic wall. PMID- 9330171 TI - Improved ultrastructural preservation of bioprosthetic tissue. AB - BACKGROUND AND AIMS OF THE STUDY: Poor ultrastructural tissue preservation of bioprosthetic heart valves is associated with a higher propensity for calcification. In spite of this realization, commercial valve fixation remains suboptimal. METHODS: In an attempt to maintain tissue integrity through improved cross-linking procedures, transmission electron microscopy and a 21-point damage score were applied to assess the ultrastructural preservation of aortic wall tissue-the main component of contemporary aortic valve bioprostheses. An ideal glutaraldehyde (GA) concentration was assessed by immediate tissue fixation at 4 degrees C comparing 0.2%, 0.5%, 0.65%, 1.0%, 2.0%, 3.0% and 4.0% GA in phosphate buffered saline (PBS). Subsequently, an optimal concentration of 3.0% GA was used to determine the effect of fixation temperature (4 degrees, 22 degrees and 37 degrees C). Finally, the superior glutaraldehyde concentration (3.0%) and cross linking temperature (4 degrees C) were used to assess tolerance towards delayed fixation. RESULTS: When different GA concentrations were used almost identical damage scores of 6.3 and 5.8 were found for 0.2% and 0.65% fixation. The first significant improvement was found at a concentration of 1.0% (score 3.3; p < 0.01) followed by a further improvement at 3.0% (score 2.6; p = 0.05). The optimal fixation temperature was 4 degrees C (3.7) with the worst results obtained at room temperature (score 9.2; p < 0.03). When fixation was delayed, the most significant damage occurred during the initial 30 min after slaughter (from 2.3 to 7.4; p < 0.02) followed by another significant deterioration between 4 and 16 h (from 5.6 to 9.7; p < 0.02). CONCLUSIONS: In summary, the prerequisites for an ideal ultrastructural preservation of bioprosthetic aortic wall tissue are immediate fixation (within 30 min), high GA concentrations (> 1.0%) and cold-temperature fixation (4 degrees C). PMID- 9330172 TI - High glutaraldehyde concentrations reduce rather than increase the calcification of aortic wall tissue. AB - BACKGROUND AND AIMS OF THE STUDY: This study was performed in order to: (i) determine whether a similar reduction of tissue calcification as seen after prolonged storage can be achieved through higher concentrations of glutaraldehyde (GA); and (ii) verify that well-preserved tissue integrity can suppress calcification. METHODS: Before fixation in 0.2% GA (PBS, 4 degrees C, seven days) porcine aortas were kept on ice for 48 h. Alternatively, tissue was immediately fixed at the abattoir in 0.2%, 1.0% or 3% glutaraldehyde (PBS, 4 degrees C, seven days). A second group of immediately fixed tissue (0.2%, 1.0%, 3.0% GA) (PBS, 4 degrees C, two days) had an interim step of L-lysine treatment (0.1M, 37 degrees C, acetic acid buffer, two days) in order to enhance cross-linking followed by warm-temperature fixation (PBS, 37 degrees C, five days). Two animal models were compared: subcutaneous implantation in rats (12 weeks) and vascular implantation in non-human primates, Chacma baboons (six weeks). RESULTS: In both animal models the highest level of calcification was found in the group with delayed fixation in 0.2% GA. In the rat model there was an inverse correlation between tissue calcification and the GA concentration used, with 3% GA-fixed tissue showing the lowest level of tissue calcium. Overall, increasing GA concentration had a significant benefit on calcification (p < 0.0001; two-factor analysis of variance). Enhancement of cross-linking with L-lysine further abrogated tissue calcium levels at all GA concentrations (p < 0.0001; two- factor analysis of variance). Although the short-term baboon model showed lower tissue calcium levels, the trend seen in the rat model was confirmed. CONCLUSIONS: Our results demonstrate the detrimental effect of delayed fixation and further suggest that, against previous beliefs, fixation at higher glutaraldehyde concentrations reduces the calcification tendency of cross-linked aortic tissue. PMID- 9330174 TI - Pericardial tissue stabilized by dye-mediated photooxidation: a review article. PMID- 9330173 TI - Glutaraldehyde detoxification of aortic wall tissue: a promising perspective for emerging bioprosthetic valve concepts. AB - BACKGROUND AND AIMS OF THE STUDY: Due to its superb crosslinking activity, glutaraldehyde (GA) is still the most widely used fixative for bioprosthetic heart valves. At the same time, however, GA is also believed to be partly responsible for tissue calcification and the lack of surface re endothelialization, both of which may contribute to valve degeneration. Although excess GA has previously been extracted from thin leaflet tissue, this treatment proved insufficient for the detoxification of thick aortic wall tissue of stentless valves or root prostheses. METHODS: In order to establish a detoxification procedure which thoroughly extracts biologically active GA from aortic wall tissue, we used a highly sensitive bioassay where endothelial cells were seeded onto glutaraldehyde-fixed aortic wall discs following various detoxification procedures. Absolute cell numbers and morphologic shape were correlated with shrinkage temperature and shrinkage extent of the tissue to determine the potential of the treatments to reverse crosslinks. To optimize treatment conditions, pH (3.2 versus 4.5), temperature (22 degrees C versus 37 degrees C) and incubation time (48 h versus one week) were varied. In order to identify an optimal detoxification agent, 12 different amino-reagents from four chemical groups were compared: low pKa aromatic amines, amino acids, low pKa N heterocyclic compounds and amino sugars. RESULTS: Amino-reagent treatment required warm temperature (37 degrees C), prolonged reaction time (one week) and a pH of 4.5 to achieve long-term cell growth on glutaraldehyde-fixed aortic wall. All 12 amino-reagents were able to detoxify aortic tissue satisfactorily; and all mildly reversed crosslinks, although there were differences between candidates. When summarized data were ranked correlating cell growth and quality with shrinkage temperature and shrinkage extent, seven reagents had a rank sum above the overall mean value, and five below with statistically significant differences between candidates. The additional stabilization of the detoxification reaction through borohydride-reduction had no further effect on tissue biocompatibility and crosslinks. CONCLUSIONS: Efficient detoxification of thick aortic wall tissue is possible if a one-week incubation in an acetic acid buffer-based amino-reagent is carried out at 37 degrees C. PMID- 9330175 TI - Time course of high-intensity transient signals in patients undergoing elective heart valve replacement: a prospective study. AB - BACKGROUND AND AIMS OF THE STUDY: This study was performed to evaluate the time course of intracranial high-intensity transient signals (HITS) in patients undergoing elective heart valve replacement. METHODS: Thirty-three patients were enrolled in this study. The examination protocol included serial (before and at one, five, 90 and 180 days after surgery) monitoring sessions with transcranial Doppler and detailed neurological examination. Monitoring was performed bilaterally over the middle cerebral arteries for one hour per session using 2 MHz probes. Microembolic signals were recognized according to standard criteria and stored on a computer for later evaluation. RESULTS: HITS prevalence increased from 3% preoperatively to 41% on the first postoperative day and remained unchanged during the postoperative period. No influence of the intensity of anticoagulation or valve type on HITS counts was evident. Unilateral monitoring provided adequate results in 83.9% of cases. CONCLUSIONS: The causative role of the valve implant in the pathogenesis of HITS appears certain, since their prevalence dramatically increases following valve implantation. Valve type, duration of valve implant or intensity of anticoagulation did not influence HITS counts. Bilateral monitoring is warranted for accurate evaluation of HITS counts in this patient group. PMID- 9330176 TI - Long-term Doppler echocardiographic follow up in normally functioning aortic St. Jude Medical prosthesis. AB - BACKGROUND AND AIMS OF THE STUDY: Mean and peak Doppler gradients remain the most frequently used parameters for follow up of prosthetic aortic valves. Gradients that deviate from baseline recordings can lead to uncertainty among physician and patient, especially if symptoms have not completely subsided after surgery, or have recurred. This study aimed to document long-term evolution of mean and peak gradients in patients with stationary clinical symptoms and signs. METHODS: Seventy-six patients (48 men, 28 women), of mean age 56.1 +/- 14.5 years (range: 23 to 82 years) who underwent St. Jude Medical bileaflet prosthesis implantation were followed up for a mean of 3.9 years (range: 1 to 7 years), both clinically and echocardiographically. Evolution of mean and peak gradients, left ventricular function, other valvular lesions and rhythm as well as adequacy of anticoagulation were examined. RESULTS: Mean gradient increased from 12.3 +/- 5.5 to 14 +/- 5.7 mmHg (p = 0.002). Mean gradient increased in 47 patients, decreased in 17 and was unchanged in 12. There was no correlation between left ventricular function, appropriate anticoagulation, left ventricular hypertrophy, age or gender with change in mean or peak gradient. Change in peak gradient correlated excellently (r2 = 0.82) with that in mean gradient. CONCLUSIONS: The range of evolution of Doppler gradients in normally functioning St. Jude Medical prostheses has been defined in this study. Slight long-term increases in mean and peak pressure gradients are normal findings and do not warrant a change in management strategy if unaccompanied by deterioration of symptoms and/or clinical signs. Although we recommend routine determination of baseline flow measurements within three months of prosthesis implantation, mean and peak gradients are adequate follow up parameters. Peak gradient correlated well with mean gradient and may be a useful adjunct for follow up in clinical practice. PMID- 9330177 TI - Early experience with the new Masters series of St. Jude Medical heart valve: in vivo hemodynamic and clinical results in patients with narrowed aortic annulus. AB - BACKGROUND AND AIMS OF THE STUDY: Aortic valve replacement in the small aortic root results in a heart-prosthesis mismatch in a significant number of patients. The new Masters series of St. Jude Medical (SJM) valves represents the company's most recent innovation, combining the beneficial Hemodynamic Plus (HP) characteristics with rotatability. Thus, this valve allows for a larger valve orifice area with an equivalent tissue annulus diameter and reduces the potential interferences of subannular tissue with leaflet mobility. METHODS: We compared prospectively the hemodynamic characteristics and the early clinical results in four groups of 25 patients each who received either the 21 Masters-HP, the 21 Standard, the 21 HP or the 23 Standard SJM valves. Patients were selected from our database and matched rigorously for age, gender, body surface area, NYHA functional class, underlying lesion, native valve opening area and left ventricular function, as well as preoperative peak and mean valve gradients. Postoperative evaluation included clinical examination and echocardiographic studies before hospital discharge and at six months. RESULTS: Short-term clinical follow up was marked by a complete absence of valve-related complications in all groups. Doppler-derived mean and maximal pressure gradients were significantly lower in the 21 HP (8.7 +/- 3.1 mmHg and 15.1 +/- 4.0 mmHg, respectively) and 21 Masters-HP groups (8.9 +/- 2.6 mm +/- Hg and 14.5 +/- 3.8 mmHg) than those in the 21 Standard group (15.1 +/- 3.2 mmHg and 22.5 +/- 6.1 mmHg; p = 0.002 and p = 0.004, respectively). These results confirm that the superior hemodynamic performance of the HP series is maintained in the Masters-HP valve, despite the introduction of a new cuff design allowing rotatability. Pressure gradients did not differ significantly between the 21 HP, the 21 Masters-HP and the 23 Standard groups. CONCLUSIONS: The hemodynamic performance of the 21 Masters-HP SJM valve corresponds closely with that of the 21 HP and 23 Standard valves and is substantially better than that of the 21 Standard valve. The Masters-HP valve will continue to reduce cardiac-prosthesis mismatch in normal-sized patients with a narrowed aortic root; its performance index is equal to that of the 21 HP valve and significantly higher than that of the 21 Standard valves. The valve will also further reduce the need for aortic annulus enlargement. PMID- 9330178 TI - Mitral valve replacement with a pulmonary autograft: the mitral top hat. AB - Despite previous unsatisfactory results with inverted pulmonary homografts in mitral valve replacement, we have rekindled our interest in this technique by the use of a pulmonary autograft with the fully flexible 'top hat' type of mounting. The surgical technique and the clinical feasibility of the operation are presented. PMID- 9330179 TI - Hufnagel procedure reconverted by endovascular leaflet dislocation and extirpation. AB - Before the development of standard aortic valve replacement, Hufnagel treated aortic insufficiency by implanting a ball-valve prosthesis in the descending thoracic aorta. In a patient who, after four major thoracic procedures, ultimately received two mechanical bileaflet valves in series (one in the ascending and one in the descending aorta), the downstream prosthesis became progressively immobilized, with total blockage of the leaflets in the open position due to insufficient transvalvular negative pressure gradient. After evaluation of this particular situation in an experimental model, we predicted blockage of the downstream prosthesis, once the ascending valve had regained normal function, and easily cleared the blockage of the distal valve by removing the two leaflets using balloon inflation. Normal circulation was clinically restored in three separate steps. (a) Normal function was re-established surgically in the ascending aorta position. (b) A second thoracotomy was avoided by endovascular dislocation of both blocked descending thoracic leaflets through endovascular balloon inflation. (c) Both leaflets embolized to the level of the left common iliac artery, where the nearly intact leaflets were removed surgically, finally creating a near-normal circulatory situation. PMID- 9330180 TI - Repair of left ventriculo-atrial fistulas due to posterior mitral annular abscesses. AB - Two cases of acute staphylococcal mitral annular abscesses resulting in transannular left ventriculoatrial fistulation are reported. Mitral annular abscess debridement and repair of the annulus, obliteration of the fistulation, and preservation of the native valve were successful in both cases. PMID- 9330181 TI - Biocompatibility of silver-modified polyester for antimicrobial protection of prosthetic valves. AB - BACKGROUND AND AIMS OF THE STUDY: The biocompatibility of a silver-coated polyethylene terephthlate (PET, polyester) fabric for the inhibition of prosthetic valve endocarditis (PVE) associated with mechanical heart valves (MHVs) was assessed. The infrequency of PVE is outweighed by mortality rates commonly exceeding 50%. These high mortality rates have been attributed to the poor effect of antibiotic therapy on colonized valves and infected myocardial tissue. Silver has been used as an antimicrobial for centuries due to its general effectiveness and relative lack of toxicity. Our previous work has shown PET polyester fabric coated with metallic silver by an ion beam-assisted deposition (IBAD) process to: (i) be effective in vitro in the inhibition of microbial attachment and colonization; (ii) be tightly adherent and low leaching; and (iii) promote tissue ingrowth and the organization of tissue pannus in a short-duration (five weeks) sheep mitral mechanical heart valve model. METHODS: This paper addresses additional biocompatibility assessment consisting of a cell compatibility assay in which serum extracts of silver-coated fabric were exposed to fibroblasts for 48 hours, after which cell viability and function were measured. The amount of silver in the extract was measured using elemental analysis techniques. RESULTS: No signs of toxicity were seen in the cells until the extract concentration reached 1200 p.p.m. Ten-week duration mechanical valve replacement studies in sheep with uncoated or coated polyester sewing cuffs showed comparable tissue ingrowth and mature pannus with a suggestion of a thinner pannus on the silver-coated fabric. Additional antimicrobial testing confirmed the effectiveness of this coating in inhibiting colonization of polyester fabric. CONCLUSIONS: These current results, together with the earlier data, suggest that IBAD silver coating on polyester facilitates healing and may provide protection against PVE. PMID- 9330182 TI - The role of IgG in type-I allergy: an unsolved problem. AB - The role of antiallergen IgG antibodies in allergy remains unclear. In this review we present evidence for and against the hypothesis that IgG (IgG4) could act as a sensitizing antibody. After considering the available data, we conclude that the possible sensitizing ability of IgG4 may depend on the nature and origin of the antibodies and cells used in the various experiments and/or is related to the differential induction of two subtypes of IgG4 (anaphylactic vs. blocking). It still remains to be settled whether the observed increase in specific IgG during immunotherapy is causally related to the relief of symptoms, or whether it merely represents an epiphenomenon due to high antigen exposure. Furthermore, the presence of IgG anti-IgE antibodies in the serum of allergic patients and their possible pathophysiological relevance is discussed in the light of recent evidence suggesting that the raised levels of these antibodies may likewise represent an epiphenomenon of the immune response. There exists a tight relationship between antiallergen IgE and IgG antibodies. This correlation has led to the suggestion that IgG determinations might be useful for diagnostic purposes. There also exists a good correlation between these antiallergen isotypes in inhibition assays. It has therefore been proposed to employ inhibition of IgG binding for the standardization of allergenic extracts. More recent studies explore the relevance of the binding affinities or avidities of allergen-specific IgG antibodies. Antibody affinity may have important repercussions with respect to the biological effects. It has been pointed out that the affinity of specific IgG subclasses in allergic patients depends, among other things, on the nature of the sensitizing allergen. Further studies on IgG binding to the so-called "major" and "minor" allergens may help clarify the role of IgG in allergic disorders. PMID- 9330183 TI - Potentiation of histamine release against inhalant allergens (Dermatophagoides pteronyssinus) with bacterial antigens in bronchial asthma. AB - In the etiopathogenesis of bronchial asthma, the important role of bacterial infection is more evident every day, favoring inflammation and obstruction, that is, triggering an asthmatic response. We gathered 36 patients diagnosed of bronchial asthma with sensitization to Dermatophagoides pteronyssinus and 32 healthy subjects. Histamine release tests against Staphylococcus aureus extract alone or together with D. pteronyssinus were performed, and the results were contrasted with or without the presence of S. aureus in the nasal secretion culture. We found histamine release against S. aureus higher than 10% only at the highest concentration (200 micrograms/ml) and significantly higher in those patients with positive nasal secretion culture. Regarding histamine release against D. pteronyssinus in the presence of S. aureus, we found a release by coincubation significantly higher than the one obtained from the addition of release against S. aureus and release against D. pteronyssinus, both in patients with negative and with positive nasal culture, at concentrations of 20 and 2 micrograms/ml. In conclusion, we observed a potentiation of histamine release against D. pteronyssinus with S. aureus extract in patients with bronchial asthma. These findings support the important role of the bacterial infection in the etiopathogenesis of bronchial asthma and the importance of a treatment against this infection. PMID- 9330184 TI - Mononuclear blood cell sulfidoleukotriene generation in the presence of interleukin-3 and whole blood histamine release in honey bee and yellow jacket venom allergy. AB - Cross-linking IgE on basophils is known to cause both sulfidoleukotriene (sLT) generation and histamine release. We recently developed an ELISA to determine sulfidoleukotriene generation by blood mononuclear cells which employs pretreatment with IL-3 to enhance leukotriene generation (cellular antigen stimulation test, CAST). Here, we compared the CAST and whole blood histamine release in response to honey bee/yellow jacket venom (BV/YJV) in 23 patients clinically suspected of type-I allergy to these venoms. Of these, 17 were diagnosed as "definitive venom allergics," defined by a positive skin test at 100 ng/ml of venom or less. The six in whom such skin reactivity was absent were labelled "suspected venom allergics." Both venoms stimulated sulfidoleukotriene generation and histamine release also from control individuals (n = 10). In patients, insect venoms generally stimulated histamine release and sulfidoleukotriene generation in excess of the mean + 3 SD of values obtained with control individuals. However, about half of the patients reacted predominantly with either histamine release or sulfidoleukotriene generation. No overall correlation was found between threshold doses necessary to stimulate sulfidoleukotriene generation (ThsLT) and histamine release (ThHist). (Linear correlation coefficients between ThsLT and ThHist were -0.02 for honey bee venom and 0.13 for yellow jacket, n = 23). Both findings are in contrast to the concept that these responses occur in parallel. From results with "definitive venom allergics," CAST sensitivity was calculated as 100% for honey bee venom and 83% for yellow jacket, and that of the histamine release assay as 62.5% for honey bee venom and 50% for yellow jacket. Specificity of the CAST was calculated as 77% for honey bee venom and 100% for yellow jacket, and that of the histamine release assay as 44% for honey bee venom and 60% for yellow jacket. Thus, CAST results are closer to skin test results than to those of the whole blood histamine release assay. PMID- 9330185 TI - Inhibition of interferon-gamma production from lymphocytes stimulated with food antigens by a beta 2-agonist, procaterol, in patients with food-sensitive atopic dermatitis. AB - Procaterol hydrochloride is a relatively new beta 2-selective agonist with a unique carbostyril nucleus. In this study, procaterol dose-dependently inhibited IFN-gamma production of peripheral blood mononuclear cells stimulated with ovalbumin in patients with hen's egg-sensitive atopic dermatitis, without inhibition of proliferative responses of peripheral blood mononuclear cells. Our results suggest that procaterol also has an effect as an immunomodulator, in addition to its effect as a beta 2-selective agonist. PMID- 9330187 TI - Allergy as an etiologic factor in nasal polyposis. AB - Allergy has been reported as an important factor in the etiology of nasal polyposis. Asthma, chronic sinusitis and aspirin hypersensitivity are frequently found together with nasal polyposis. Total IgE, RAST for specific IgE and skin prick test were used to investigate the incidence of allergy in 95 patients with nasal polyposis. In addition, histopathologic appearance of polyp tissue was examined in 21 patients after polypectomy and compared in allergic and nonallergic groups. IgG subclass levels were also measured to detect if there were any changes. Mean serum IgE level was found to be higher in the patient group and the skin prick test (SPT) was positive in 66.3% of patients. On the basis of positive SPT and serum RAST results, 45.2% of all patients with nasal polyposis were defined as allergic. Both total IgE and IgG4 were detected at increased levels in the SPT-positive group. These findings suggest that an IgE mediated mechanism may be present in a subpopulation of patients with nasal polyposis. PMID- 9330186 TI - Atopy and bronchial hyperresponsiveness in pure extrinsic childhood asthma. AB - Nonspecific bronchial hyperresponsiveness and atopy are considered risk factors in the development of asthma. Bronchial responsiveness to allergens could be the most important factor in extrinsic asthma. The trial was designed to investigate the role of specific and nonspecific bronchial responsiveness and atopy in a pure model of extrinsic asthma in children. One hundred and thirty-seven patients with pollen allergy were evaluated. Twenty children with allergy to grass pollen (Lolium perenne) alone, with symptoms only in the grass pollen season, were selected. Their score of symptoms, airway responsiveness to methacholine in and out of season, airway responsiveness to Lolium perenne out of season, and total and specific IgE were assessed. Twelve were male and eight female. Mild asthma was observed in 14, and moderate asthma in six. Age of onset of symptoms ranged from three to 13 years of age. Significant seasonal increase in airway responsiveness to methacholine was found (p = 0.002). Specific bronchial challenge test was positive in all patients. Lolium pernne PD20 ranged from 2.3 to 155.5 inhalation units. An inverse association between age of onset of symptoms and severity of asthma was shown (p = 0.001). Increase in nonspecific bronchial responsiveness was related to the appearance of symptoms during the spring, but it showed no relationship to the severity of symptoms. Severity of asthma during the spring correlated with the intensity of allergen airway responsiveness (p = 0.02). Levels of total and specific IgE were not related to the degree of specific or nonspecific airway responsiveness. Severity of extrinsic childhood asthma is determined by bronchial response to allergens. Bronchial hyperresponsiveness to methacholine during the spring can be the consequence of environmental exposure to allergens. The intensity of airway responsiveness to methacholine has no predictive value in the severity of pure extrinsic childhood asthma. PMID- 9330188 TI - Measurement of allergen-induced skin reactions by computerized dynamic telethermography (CDTT). AB - Cutaneous thermic changes induced by a skin prick test reaction were measured by an infrared thermography camera (computerized dynamic telethermography, CDTT). Changes in skin temperature (T degree) detected by CDTT were compared with the mean diameter of allergen-induced skin reactions. Cutaneous thermic increase detected by CDTT correlated well with the mean wheal diameter measured in millimeters (r = 0.938, p < 0.001). Average coefficient of variation for repeated CDTT measurements was 4.6%. CDTT provides a reproducible and precise method for measuring allergen-induced skin reactions. Moreover, the continuous recording of the skin temperature represents an additional parameter for the quantification of wheal reactions. PMID- 9330189 TI - IgE antibodies to aeroallergens in allergic children in Sao Paulo, Brazil. AB - The sensitization to inhaled allergens was studied in 80 Brazilian children and 12 controls, aged 6 to 16 years. We performed skin tests with Alternaria alternata, cat, dog, Lolium perenne, grasses and the following domestic mites: Blomia tropicalis, Dermatophagoides pteronyssinus, Dermatophagoides farinae, Lepidoglyphus destructor, Chortoglyphus arcutus and Aleuroglyphus ovatus. The frequencies of positive skin tests (mean diameter > or = 23 mm) were respectively 15%, 11%, 11%, 6%, 7%, 95%, 92%, 88%, 76%, 75% and 71%. The mean diameter of the wheal to domestic mites was higher to D. pteronyssinus than to B. tropicalis, L. destructor, A. ovatus and C. arcuatus (p < 0.05). The skin test results to domestic mites by regression analysis revealed highly significant correlation (p < 0.0001), suggesting considerable cross-reactivity between them. Specific IgE to each mite was determined and the results expressed as the mean of the percentage of total counts bound (% TCB). We had the highest levels of % TCB to D. farinae, followed by D. pteronyssinus, E. maynei, B. tropicalis, C. arcuatus, L. destructor and A. ovatus. The Brazilian children with asthma had more positive skin tests to domestic mites than to other inhalant allergens, and based on previous studies performed in Sao Paulo that have shown D. pteronyssinus and B. tropicalis as the most prevalent mite species in house dust, we assumed that IgE Ab to D. farinae reflects cross-reactivity with D. pteronyssinus in most cases. The observed high levels of IgE Ab to domestic mites stress the importance of environmental avoidance measures, decreasing the rate of sensitization or perhaps the development of symptoms of asthma in genetically predisposed children. PMID- 9330190 TI - Asthma mortality in Latin America. AB - There are not enough data concerning asthma mortality in Latin America. The Latin American Society of Allergy and Immunology coordinated this project to provide reliable data for gaining knowledge about our present situation, which is a condition indispensable to changing it. The following countries participated in this study: Argentina, Brazil, Chile, Colombia, Costa Rica, Cuba, Mexico, Paraguay, Peru, Uruguay and Venezuela. A uniform protocol was designed in Santa Fe, Argentina. Asthma mortality rates were analyzed in accordance with two variables: age-adjusted rates (5-34) and total death rates. The total population studied was 107, 122, 529 inhabitants. The highest death rates were found in Uruguay and Mexico (5.63), and the lowest in Paraguay (0.8) and Colombia (1.35). Age-adjusted (5-34) rates were higher in Costa Rica (1.38) and lower in Chile (0.28). Regarding sex, the analysis of the information provided by seven countries showed a predominance of females (51.8%) over males (48.18%). In the southern Latin American countries such as Chile, Uruguay, Paraguay and Argentina, which have marked climatic differences, deaths occurred mainly in the winter. It is important to emphasize that, in most countries, deaths from asthma occurred at home: Chile (60.7%), Argentina (63.4%) and Paraguay (88%). However, in Uruguay, 58.6% occurred during hospitalization. Mortality rates from bronchial asthma are high in most of the Latin American countries studied, even though further studies are needed. Asthma is a serious global health problem. People of all ages in countries throughout the world are affected by this chronic airway disorder that can be severe and sometimes fatal. The health ministries of each country do not believe asthma is a significant issue. Therefore, we should provide them with sound epidemiological studies to convince them to change their attitude toward this disease. PMID- 9330192 TI - Allergic contact dermatitis from tromantadine. AB - Tromantadine is a topical antiviral agent derived from amantadine. In previous reports, 5% of treated patients developed contact allergy. We report the case of a 28-year-old woman who had a sudden worsening of a lip herpes after treatment with a tromantadine ointment, with strong positive patch testing to this substance. The purpose of this paper is to call attention to the fact that tromantadine is still widely used in several European countries as a second-line therapy for lip herpes, and therefore, new cases of sensitization to tromantadine are likely to occur. PMID- 9330191 TI - Monocyte chemotactic and activating factor/monocyte chemoattractant protein (MCAF/MCP-1) in bronchoalveolar lavage fluid from patients with atopic asthma and chronic bronchitis. AB - Monocyte chemotactic and activating factor/monocyte chemoattractant protein (MCAF/MCP-1) is a member of the beta (C-C) subfamily of chemokines. The biological roles played by MCAF/MCP-1 in a number of inflammatory and noninflammatory diseases states is not well known. Several studies have confirmed that inflammation is present in the airways of subjects with atopic asthma and with chronic bronchitis. Analysis of bronchoalveolar lavage fluid (BALF) is an effective method of sampling lower respiratory tract inflammation. The aim of this study was to examine associations among MCAF/MCP-1, BALF cells and spirometry parameters and bronchial hyperresponsiveness in patients with atopic asthma and chronic bronchitis. Twenty patients with atopic asthma, 10 patients with chronic bronchitis and 10 patients of the control group, took part in this study. An ELISA test was used to assess MCAF/MCP-1 in BALF. The levels of MCAF/ MCP-1 (mean +/- SEM) were 150 +/- 18.6 pg/ml in patients with atopic asthma, 320 +/- 39.7 pg/ml in chronic bronchitis and 74.9 +/- 3.3 pg/ml in the control group (p < 0.05). When all patients with disease were considered, there was negative correlation with FEF50 (Kendall's correlation coefficient = - 0.4; p < 0.01). Regression analysis has shown that a level of MCAF/MCP-1 over 100 pg/ml was correlated with duration of illness (Pearson's correlation coefficient = 0.7; p < 0.02). In conclusion, MCAF/MCP-1 probably possesses proinflammatory properties in atopic asthma and chronic bronchitis. The elevated level of this chemokine may support the clinical suspicion of specific diagnosis. PMID- 9330193 TI - Peter Howard Elworthy (1933-1995): a biographical note. AB - Peter Elworthy had a considerable influence on pharmaceutical science, education and practice, in the UK. He died in December 1995 at the early age of 62, but he had retired from full time academic work from his post as Professor of Pharmacy and Head of the Department of Pharmacy in the University of Manchester twelve years earlier. Concerned about his health but also not a little disillusioned by the multiple pressures placed even then on senior academics, he foresaw the era of cuts and the central oversight and restrictions. His heart was elsewhere. In his 1976 Harrison Memorial Lecture (reproduced in this issue) he said: "Looking back, I have the feeling of having been very lucky. 'Much have I travell'd in the realms of gold, and many goodly states and kingdoms seen.' My main interest has been in the phenomenon of micellization. The subject has been of absorbing interest, and continually shows new bright facets which lead to new scientific advances. Travelling in the realms of gold has nothing to do with gold, but to me it means travelling in sunlight, which illuminates things brightly, and makes visible new facts, which have been invisible before. Occasionally dark clouds form; they are the disappointments and frustrations, but we need them in order to be able to recognize the sunlight by contrast." PMID- 9330194 TI - Dehydrated elephants and other matters. 1971. PMID- 9330195 TI - The increasingly clever micelle. 1976. PMID- 9330196 TI - An interfacial tension model of the interaction of water-soluble polymers with phospholipid composite monolayers. AB - The axi-symmetric drop-shape analysis-pendant drop technique has been used to measure interfacial tension at the chlorobenzene-water interface in the presence of adsorbed films of dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), DMPC-cholesterol, DPPC-cholesterol, DMPC cholesterol-dicetyl phosphate (DCP) and DPPC-cholesterol-DCP. A surface-pressure function, pi * = pi lipid-polymer -pi lipid (where pi lipid is the surface pressure of the mono-layer without polymer and pi lipid-polymer is the surface pressure of the lipid mono-layer and adsorbed polymer at equilibrium at the chlorobenzene-water interface) was used to characterize the interaction of eight water-soluble polymers with the lipid films. The equation, delta pi * = pi II*-pi I* (where the subscripts II and I denote the higher and lower lipid composites, respectively) was used to determine the differential effect of cholesterol and DCP on mono-layer characteristics in the presence of 1% w/v polymer. Cholesterol or polymer individually condensed DMPC films and expanded DPPC films. However, composite films of DMPC-cholesterol-DCP and carboxymethylchitin (CM-chitin), poly(acrylic acid) (PAA) or poly(vinyl alcohol) (PVA) were more expanded than DMPC films whereas composite films of DPPC were neither more condensed nor expanded than DPPC films. A polymer impact ratio, P* = pi lipid-polymer/pi lpolymer was calculated and the polymers were ranked in order of their impact on the lipid film. PVA and polysaccharides gave low and high P* values, respectively, corresponding to high and low levels of film interaction, whereas PAA and hydrophobized polysaccharides gave intermediate values, indicating their affinity for and penetration of interfacial films with little disruption of the mono-layer. The results show that measurement of interfacial pressures at the chlorobenzene-water interface might be advantageous for evaluating the action of polymers on biological membranes. PMID- 9330197 TI - Variation in cetostearyl alcohol and lecithin from different sources: evaluation by dielectric analysis. PMID- 9330198 TI - Aggregation and surface properties of synthetic double-chain non-ionic surfactants in aqueous solution. AB - Double-chain non-ionic surfactants have been synthesized with the general formula 2CnMPEG750, where n is the number of carbons in the alkyl chains and 750 is the molecular weight of the monomethoxypolyoxyethylene (MPEG) head group. The aggregation and surface properties in dilute aqueous solution (< 1.0% w/w) of surfactants with n = 8, 10 and 12 have been determined. Each of the surfactants formed clear, foaming, non-birefringent solutions. Total-intensity light scattering indicated the presence of micelles with aggregation numbers of 36, 68 and 74 for the 2C8, 2C10 and 2C12 surfactants, respectively, whereas photon correlation studies yielded radii, assuming spherical aggregates, of 59-103 A. Surface-tension measurements gave critical micelle concentrations for the surfactants in the range 1.15-7.24 x 10(-6) M; these, as expected, decreased when the number of carbon atoms in the alkyl chains was increased. Such studies are important in the design of new surfactants as vehicles for drug delivery because it is imperative to be able to predict the type of aggregate formed by a surfactant. PMID- 9330199 TI - The influence of gamma irradiation on the physicochemical properties of a novel triblock copolymer of epsilon-caprolactone and ethylene oxide. AB - A novel triblock copolymer of epsilon-caprolactone (CL) and ethylene oxide (E), CL6E90CL6, intended for use in implantable drug-delivery systems, has been subjected to gamma irradiation, in the solid state and in aqueous solution, under different controlled environmental conditions, to assess its stability to a radiation sterilization process. When copolymer matrices were irradiated with doses of irradiation up to 72 kGy in the presence of oxygen, negligible changes were observed in the molar mass, molecular mobility (assessed by pulsed nuclear magnetic resonance spectroscopy) and thermal properties. However, irradiation of matrices in the absence of oxygen (anoxia) induced the formation of cross-links, as indicated by a reduction in the molecular mobility of the copolymer, but without affecting its molar mass and thermal properties. Gamma irradiation of aqueous solutions of CL6E90CL6 in the presence of oxygen induced random polymer chain scission, as evidenced by a reduction in the molar mass, and the formation of a distribution of copolymer chain lengths in solution. Nuclear magnetic resonance relaxation studies showed that irradiation of solutions of CL6E90CL6 at concentrations greater than 4% w/v under anoxic conditions with doses of 54 kGy produced polymer gels with a network structure. These differences in the effects of gamma irradiation on the physicochemical properties of CL6E90CL6 might be germane to the method selected for sterilization of the polymer before its use in implantable drug-delivery systems. PMID- 9330200 TI - Polyhedral non-ionic surfactant vesicles. AB - Large polyhedral (2-10 microns) non-ionic surfactant vesicles (niosomes) formed from mixtures of a hexadecyl diglycerol ether (C16G2), a cholesteryl poly-24 oxyethylene ether (solulan C24) and a low level of cholesterol are being investigated as slow-release systems for ophthalmic, subcutaneous or intramuscular administration. The phase-diagram of this three-component system has been constructed and these polyhedral vesicles are found to be in the gel (L beta) phase. Confocal laser-scanning microscopy was used to confirm the complex morphology of these vesicles. The thermo-responsive nature of release of entrapped carboxyfluorescein and nicotinamide adenine dinucleotide has been studied; release is increased with increase in temperature (37 degrees C) even though the polyhedral vesicles still maintain their polyhedral shape at this temperature. The results indicate that the thermo-responsive features of the niosomes are a result of reversible changes in bi-layer permeability caused by temperature-mediated alteration in the membrane-packing characteristics of the polyethoxylated cholesterol ether. PMID- 9330201 TI - The effect of monomers and of micellar and vesicular forms of non-ionic surfactants (Solulan C24 and Solulan 16) on Caco-2 cell monolayers. AB - Measurements of transepithelial electrical resistance (TEER), the MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) test and monitoring of poly(ethylene glycol) (PEG) transport have been used to study the effects of the non-ionic surfactants Solulan C24 and Solulan 16, either free in solution or as an integral part of niosome bi-layers, on intestinal epithelial cells from man (Caco-2 cell monolayers). The effects on epithelial integrity and on the transport of the hydrophilic drug metformin depend on the concentration of the surfactants. At concentrations above 1% the effect on TEER of the surfactant in niosomal form and free in solution were equivalent whereas cell viability was preserved to a higher concentration of Solulans when the Solulans were present in the niosomal form. It was concluded that the toxic effect of niosomes arises from free surfactant present in the niosome suspension. PMID- 9330202 TI - Influence of hydroxypropyl beta-cyclodextrin on the stability of benzylpenicillin in chloroacetate buffer. AB - Hydroxypropyl beta-cyclodextrin (HP beta CyD) has been shown to stabilize a wide variety of chemically distinct pharmaceutical entities through inclusion-complex formation between drug and cyclodextrin. The effect of HP beta CyD on the acid catalysed hydrolysis of benzylpenicillin (penicillin G) was evaluated in chloroacetate buffer at pH 2.20. At penicillin G: cyclodextrin molar concentration ratios from 1:1 to 1:10, HP beta CyD effected stabilization of penicillin G by 1.56- to 5.21-fold. At all temperatures, the observed first-order rate constant (kobs) values assumed a non-linear, Michaelis-Menten type decrease as a function of increasing HP beta CyD concentration. Degradation of penicillin G complexed with HP beta CyD (penicillin G-HP beta CyD), was approximately ninefold slower than uncomplexed penicillin G. The proportion of penicillin G degrading in either of these forms was, in turn, determined by the equilibrium constant for the complexation. The apparent thermodynamic and activation parameters for the complexation between penicillin G and HP beta CyD have also been evaluated. The negative standard enthalpy change (delta H degrees) for the complexation implied that the penicillin G-HP beta CyD complex would be predisposed towards enhanced stability, and thus the kobs value for the hydrolysis of penicillin G decreased with reduction of temperature in these systems. The lack of difference between the enthalpies of activation (delta H ++) for the hydrolysis of uncomplexed and complexed penicillin G seemed to be compensated by the significant difference between the entropies of activation (delta S ++) for these hydrolytic reactions. The results indicate that HP beta CyD represents a viable means of stabilization of penicillin G solutions at the pH employed in this study. PMID- 9330203 TI - Studies on amidated pectins as potential carriers in colonic drug delivery. AB - Amidated pectins have been assessed, in-vitro, for their potential value in colonic drug delivery. The monitoring of the release of a model soluble drug, paracetamol, gives a sensitive indication of the behaviour of the pectins under simulated gastrointestinal conditions. Inclusion of calcium as a cross-linking agent increased the viscosities of amidated pectin gels to a maximum value. Further addition of calcium reduced gel viscosity and for pectin with a high extent of amidation this led to a reduction in drug release. Release was faster for pectin with less amidation in the presence of calcium; this could be related to matrix erosion. The results of the study suggest that the materials might be of value in colonic delivery either alone or in combination, possibly in the form of a coating. PMID- 9330225 TI - Limited proteolysis of angiogenin by elastase is regulated by plasminogen. AB - Human neutrophil elastase cleaves angiogenin at the Ile-29/Met-30 peptide bond to produce two major disulfide-linked fragments with apparent molecular weights of 10,000 and 4000, respectively. Elastase-cleaved angiogenin has slightly increased ribonucleolytic activity, but has lost its ability to undergo nuclear translocation in endothelial cells, a process essential for angiogenic activity. Cleavage appears to alter the cell-binding properties of angiogenin, despite the fact that it occurs some distance from the putative receptor-binding site, since the elastase-cleaved protein fails to compete with its native counterpart for nuclear translocation in endothelial cells. Plasminogen specifically accelerates elastase proteolysis of angiogenin. It does not enhance elastase activity toward ribonuclease A or the synthetic peptide substrate MeOSuc-Ala-Ala-Pro-Val-pNA. Plasminogen-accelerated inactivation of angiogenin by elastase might be a significant event in the process of angiogenin-induced angiogenesis since (i) angiogenin and plasminogen circulate in plasma at high concentrations, (ii) angiogenin, especially when bound to actin, activates tissue plasminogen activator to generate plasmin from plasminogen, and (iii) elastase cleaves plasminogen to produce angiostatin, a potent inhibitor of angiogenesis and metastasis. Interrelationships among angiogenin, plasminogen, plasminogen activators, elastase, and angiostatin may provide a sensitive regulatory system to balance angiogenesis and antiangiogenesis. PMID- 9330227 TI - Artificial neural network method for predicting the specificity of GalNAc transferase. AB - The specificity of GalNAc-transferase is consistent with the existence of an extended site composed of nine subsites, denoted by R4, R3, R2, R1, R0, R1', R2', R3', and R4', where the acceptor at R0 is either Ser or Thr to which the reducing monosaccharide is anchored. To predict whether a peptide will react with the enzyme to form a Ser- or Thr-conjugated glycopeptide, a neural network method- Kohonen's self-organization model is proposed in this paper. Three hundred five oligopeptides are chosen for the training site, with another 30 oligopeptides for the test set. Because of its high correct prediction rate (26/30 = 86.7%) and stronger fault-tolerant ability, it is expected that the neural network method can be used as a technique for predicting O-glycosylation and designing effective inhibitors of GalNAc-transferase. It might also be useful for targeting drugs to specific sites in the body and for enzyme replacement therapy for the treatment of genetic disorders. PMID- 9330226 TI - Intrinsic fluorescence in endoglucanase and cellobiohydrolase from Trichoderma pseudokiningii S-38: effects of pH, quenching agents, and ligand binding. AB - To gain further insight into the difference in substrate specificity between endoglucanase and cellobiohydrolase, the intrinsic fluorescence properties of cellobiohydrolase I (CBH I) and endoglucanase I (EG I) from Trichoderma pseudokiningii S-38 were investigated. The results for the spectral characteristics, ligand binding and fluorescence quenching suggest that the fluorescence of two enzymes comes from tryptophan residues, and that tryptophan residue(s) may be involved in the function of the two enzymes. The results also suggest that the binding tryptophan in EG I may be more exposed to solvent than that in CBH I. This interpretation is supported by the observations that the effects of pH upon the fluorescence of EG I are greater than that of CBH I; spectral shifts are different in EG I and CBH I under various conditions, and fluorescence lifetime changes caused by cellobiose binding are larger for EG I than for CBH I. PMID- 9330224 TI - Preformed GroES oligomers are not required as functional cochaperonins. AB - We have previously shown that the C-terminal sequence of GroES is required for oligomerization [Seale and Horowitz (1995), J. Biol. Chem. 270, 30268-30270]. In this report, we have generated a C-terminal deletion mutant of GroES with a significantly destabilized oligomer and have investigated its function in the chaperonin-assisted protein folding cycle. Removal of the two C-terminal residues of GroES results in a cochaperonin [GroESD(96-97)] that is monomeric at concentrations where GroES function is assessed. Using equilibrium ultracentrifugation, we measured the dissociation constant for the oligomer monomer equilibrium to be 7.3 x 10(-34)M6. The GroESD(96-97) is fully active as a cochaperonin. This mutant is able to inhibit the ATPase activity of GroEL to levels comparable to wild-type GroES. It is also able to assist the refolding of urea-denatured rhodanese by GroEL. While GroESD(96-97) can function at levels comparable to wild-type GroES, higher concentrations of mutant are required to produce the same effect. These results support the idea that the performed GroES heptamer is not required for function, but they suggest that the oligomeric cochaperonin is most efficient. PMID- 9330229 TI - PFDB: a protein families database for Macintosh computers. The effectiveness of its organization in searching for protein similarity. AB - A protein sequence database (PFDB) containing about 11,000 entries is available for Macintosh computers. The PFDB can be easily updated by importing sequences from the PIR collection through the internet. The most important feature of the database is its organization in families of closely related sequences, each family being characterized by its average dipeptide composition [Petrilli (1993), Comput. Appl. Biosci. 2, 89-93]. This allows one to perform a rapid and sensitive protein similarity search by comparing the precalculated family dipeptide composition with that of the query sequence by a linear correlation coefficient. An example of an application in which a new protein was classified by using a sequence of a fragment just 19 residues long is reported. PMID- 9330228 TI - Botulinum neurotoxin type A: limited proteolysis by endoproteinase Glu-C and alpha-chymotrypsin enhanced following reduction; identification of the cleaved sites and fragments. AB - Botulinum neurotoxin (NT) serotype A is a approximately 150-kDa dichain protein. Posttranslational nicking of the single-chain NT (residues Pro 1-Leu 1295) by the protease(s) endogenous to Clostridium botulinum excises 10 residues, leaving Pro 1-Lys 437 and Ala 448-Leu 1295 in the approximately 50-kDa light (L) and approximately 100-kDa heavy (H) chains, respectively, connected by a Cys 429-Cys 453 disulfide and noncovalent bonds [Krieglstein et al. (1994), J. Protein Chem. 13, 49-57]. The L chain is a metalloprotease, while the amino- and carboxy terminal halves of the H chain have channel-forming and receptor-binding activities, respectively [Montecucco and Schiavo (1995), Q. Rev. Biophys. 28, 423 472]. Endoproteinase Glu-C and alpha-chymotrypsin were used for controlled digestion at pH 7.4 of the approximately 150-kDa dichain NT and the isolated approximately 100-kDa H chain (i.e., freed from the L chain) in order to map the cleavage sites and isolate the proteolytic fragments. The dichain NT appeared more resistant to cleavage by endoproteinase Glu-C than the isolated H chain. In contrast, the NT with its disulfide(s) reduced showed rapid digestion of both chains, including a cleavage between Glu 251 and Met 252 (resulting in approximately 30- and approximately 20-kDa fragments of the L chain) which was not noted unless the NT was reduced. Interestingly, an adjacent bond, Tyr 249-Tyr 250, was noted earlier [DasGupta and Foley (1989), Biochimie 71, 1193-1200] to undergo "self-cleavage" following reductive separation of the L chain from the H chain. The site Tyr-Tyr-Glu-Met (residues 249-252) appears to become exposed following reduction of Cys 429-Cys 453 disulfide. Identification of Glu 669-Ile 670 and Tyr 683-Ile 684 as protease-susceptible sites demonstrated for the first time that at least two peptide bonds in the segment of the H chain (residues 659 684), part of which (residues 659-681) is thought to interact with the endosomal membranes and forms channels [Oblatt-Montal et al., (1995), Protein Sci, 4, 1490 1497], are exposed on the surface of the NT. Two of the fragments of the H chain we generated and purified by chromatography are suitable for structure-function studies; the approximately 85- and approximately 45-kDa fragments beginning at residue Leu 544 and Ser 884, respectively (both extend presumably to Leu 1295) contain the channel-forming segment and receptor-binding segments, respectively. In determining partial amino acid sequences of 10 fragments, a total of 149 amino acids in the 1275-residue NT were chemically identified. PMID- 9330230 TI - A model for the unusual kinetics of thermal denaturation of rubredoxin. AB - The thermal denaturation of the simple, redox-active iron protein rubredoxin is characterized by a slow, irreversible decay of the characteristic red color of the iron center at elevated temperatures in the presence of oxygen at pH 7.8. The denaturation rate is essentially constant and the time period for complete bleaching is nearly independent of protein concentration. These two characteristics of the kinetics can be fit by a simple self-catalyzed kinetics model consisting of the combination of a first-order decay and catalysis by some product of that decay, i.e., dP/dt = k1[A] + (k2[P][A])/(K(m) + [A]), where A is native rubredoxin, P, is unspecified product, k1 is a first-order rate constant, and k2 and K(m) are the catalytic constants. In order for the second term to be of this simple form over the full course of a decay, the model must include the condition that the reaction is effectively irreversible. This model has properties which suggest other biological roles in regulation (changes in k1 or k2 can dramatically modulate the kinetics), in timing (titer-independent fixed reaction time), and in self-activation reactions. At one extreme (k1 >> k2) the kinetics becomes exponential, but at the other extreme (k2 >> k1) they show a dramatic and rapid terminal increase after a lag period. Some obvious possible roles in the kinetics of programmed cell death, prion disease, and protease autoactivation are discussed. PMID- 9330231 TI - Neuropsychiatry--the mind embrained? PMID- 9330232 TI - Traumatic brain injury and psychiatry. PMID- 9330233 TI - The relationship between infection and fatigue. PMID- 9330234 TI - Attitudes toward motherhood in postnatal depression: development of the Maternal Attitudes Questionnaire. AB - This article describes the development of the Maternal Attitudes Questionnaire (MAQ), a 14-item self-report instrument measuring cognitions relating to role change, expectations of motherhood, and expectations of the self as a mother in postnatal women. This questionnaire was found to have good test-retest and internal reliability. In a large sample of women (n = 483) at 6-8 weeks postpartum, scores on the questionnaire were highly correlated with scores on the Edinburgh Postnatal Depression Scale (EPDS) and the Revised Clinical Interview Schedule (CIS-R). Cluster analysis demonstrated that, among depressed women with similar symptom scores on the CIS-R, the MAQ discriminated a group with low MAQ scores and a group with high MAQ scores. This finding supports the hypothesis that women who are depressed postnatally are cognitively heterogeneous; such differences may be important in understanding the etiology and determining the treatment of postnatal depression. PMID- 9330235 TI - Sleep disruption and mood changes associated with menopause. AB - This study examined the sleep and mood differences between premenopausal and perimenopausal women matched for age and sociodemographic variables. Wrist actigraphy, Profile of Mood State (POMS), State-Trait Anxiety Inventory (STAI), a sleep questionnaire, and responses to a sleep diary were recorded for a period of 1 week. It was found that the sleep disruption of perimenopausal subjects was significantly greater than that of the premenopausal group (p < 0.05). Overall, the perimenopausal group demonstrated a significant increase in sleep disruption and mood alterations when compared with the premenopausal group. Actigraphic data showed that perimenopausal subjects experienced longer and more numerous arousals resulting in significantly less sleep (p < 0.05). In addition, perimenopausal subjects scored significantly higher (p < 0.05) on the STAI and significantly lower on the Vigor subscale of the POMS (p < 0.01) than premenopausal subjects. Correlational analyses indicated that sleep and mood changes were significantly related in the perimenopausal group, but not in the premenopausal group. Taken together, these results suggest that the mood changes experienced by the perimenopausal group may be mediated by sleep disruption. PMID- 9330236 TI - The chronic fatigue syndrome and hyperventilation. AB - Chronic fatigue syndrome (CFS) is characterized by severe fatigue, lasting for at least 6 months, for which no somatic explanation can be found. Because hyperventilation can produce substantial fatigue, it seems worthwhile to investigate the relationship between it and CFS. It might be hypothesized that hyperventilation plays a causal or perpetuating role in CFS. CFS patients, non CFS patients known to experience hyperventilation, and healthy controls were compared on complaints of fatigue and hyperventilation. CFS patients and non-CFS patients known to experience hyperventilation offered substantial complaints of fatigue and hyperventilation, both to a similar degree. Physiological evidence of hyperventilation was found significantly more often in CFS patients than in healthy controls. However, no significant differences between CFS patients with and CFS patients without hyperventilation were found on severity of fatigue, impairment, number of complaints, activity level, psychopathology, and depression. It is concluded that hyperventilation in CFS should probably be regarded as an epiphenomenon. PMID- 9330237 TI - Chronic posttraumatic stress disorder and chronic pain in Vietnam combat veterans. AB - A study was conducted to investigate chronic pain patterns in Vietnam veterans with posttraumatic stress disorder (PTSD). Combat veterans with PTSD completed standardized PTSD severity, pain, somatization, and depression measures. Of 129 consecutive out-patient combat veterans with PTSD, 80% reported chronic pain. In descending order were limb pain (83%), back pain (77%), torso pain (50%), and headache pain (32%). Compared to PTSD combat veterans without chronic pain, PTSD veterans who reported chronic pain reported significantly higher somatization as measured by the Minnesota Multiphasic Inventory 2 hypochondriasis and hysteria subscales. In the sample of 103 combat veterans with PTSD and chronic pain, MMPI 2 hypochondriasis scores and B PTSD symptoms (reexperiencing symptoms) were significantly related to pain disability, overall pain index, and current pain level MMPI 2 hypochondriasis and depression scores were also significantly related to percent body pain. These results are discussed in the context of current conceptualizations of PTSD. PMID- 9330238 TI - Apparent poisoning by wood preservatives: an attributional syndrome. AB - Recent reports have pointed to an increased number of patients presenting with multisystem symptoms which they attribute to chemical exposures or to heightened chemical sensitivity. Twenty patients exposed to wood preservative products, who attended a joint toxicology and psychiatric clinic, were reviewed by a retrospective case note analysis. Thirteen patients attributed their symptoms to the wood preservative soon after the exposure, and seven patients developed the attribution only at a later date. Reported symptoms referred to all body systems, but there were few physical signs. Clinical findings suggest that the acute symptoms were consistent with the expected toxic effects, but the chronic symptoms could not be explained physically. Patient's beliefs about chemical poisoning could be understood as arising in the context of an attributional process, representing a sociopsychosomatic syndrome precipitated by wood preservative exposure. Patient management included a discussion of findings from assessments, published information, along with counseling where appropriate. Follow-up information from their general practitioners indicated a possible improvement in 50% of patients. PMID- 9330239 TI - Does short-term group therapy affect unexplained medical symptoms? AB - This paper describes an intensive time-limited group-therapy program conducted in a busy surgical clinic by two nursing staff with the support of a consultant psychiatrist. Nineteen patients with chronic idiopathic facial pain were recruited to the study and underwent weekly group therapy over 8 weeks. At the end of the study period, results showed decreases in pain, anxiety and depression scores, along with an improvement in the patients coping skills. The findings support the use of group psychological interventions undertaken by appropriately trained nursing staff in reducing symptoms associated with chronic idiopathic facial pain. PMID- 9330240 TI - Coping and other predictors of outcome in chronic fatigue syndrome: a 1-year follow-up. AB - In this prospective study, 137 patients with chronic fatigue syndrome were followed-up at a 1-year interval to determine factors relating to outcomes. Nearly two thirds reported an improvement on direct ratings of change. In analyses with fatigue and functional impairment at follow-up as the criteria, and controlling for earlier status, poorer outcomes were predicted by illness duration, subjective cognitive difficulty, and somatic symptoms; there was no influence of anxiety, depression, or general emotional distress. Fatigue was also predicted by information-seeking, and impairment by behavioral disengagement and a low internal locus of control. The belief that one's actions can influence outcomes modified the relationship between illness accommodation and both fatigue and impairment; adverse outcomes were associated with accommodating to illness only in the context of lower levels of perceived control. Thus, it is suggested that interventions that either discourage avoidance of activity or enhance perceived control could benefit the course of the illness. PMID- 9330241 TI - Abbreviated cognitive test for delirium. AB - The cognitive test for delirium (CTD) was recently developed to identify delirium in an intensive care unit (ICU) setting. Stepwise discriminant analyses using the original validation sample indicated that a total score formed by summing only two of the nine content scores (visual attention span and recognition memory for pictures) maintained good reliability (coefficient alpha = 0.79) and the ability to discriminate delirium from dementia, schizophrenia, and depression (p < 0.0001) and delirium from moderate to severe dementia (p < 0.0002). This abbreviated version of the CTD is more practical for use by ICU clinicians. PMID- 9330242 TI - Conversion disorders in Nottingham: alive, but not kicking. AB - A postal request to general practitioners in a catchment area (population 37,000) identified 18 patients fulfilling criteria for conversion disorder (age range 26 74 years; mean age of onset of first episode 38 years). There was a female preponderance, with two patients from ethnic minority groups. There was a temporal correlation with stress in 13 cases (72%) and a history of sexual abuse in 5 cases (28%). Three clinical groups were identified: acute onset with good premorbid functioning and full recovery; conversion symptoms as part of polysymptomatic presentation, with fluctuating course; and chronic, severely disturbed individuals with a past history of sexual abuse. One third of cases were referred to psychiatrists. There was no case of a missed organic disorder. The details of one case are discussed. PMID- 9330243 TI - Burn injuries, psychiatric disorders and length of hospitalization. AB - The purpose of this work was to investigate whether differences in length of hospitalization exist between burn patients with and without psychiatric comorbidity and, if so, why. The method undertaken was a retrospective file study from 1981 to 1995. Psychiatric patients with accidental burns are hospitalized longer than nonpsychiatric patients. However, psychiatric patients with self inflicted burns are not hospitalized longer (after correction for the size of burn). The most likely reason for this is an excess of substance-dependent patients in the group of psychiatric patients with accidental burns. The often reported longer hospitalization of psychiatric patients may be restricted to substance-dependent patients and may be mediated by poorer physical condition. PMID- 9330244 TI - Effects of lidocaine on cytosolic pH regulation and stimulus-induced effector functions in alveolar macrophages. AB - Local anesthetics influence a variety of stimulus-induced effector functions in leukocytes. The present study determined the effects of lidocaine on intracellular pH (pHi) regulation, superoxide production, and tumor necrosis factor-alpha (TNF-alpha) release in alveolar macrophages (m phi). Resident m phi were obtained by bronchoalveolar lavage of rabbits. The cells were subjected to an intracellular acid load, and subsequent pHi recovery was followed in the presence or absence of bafilomycin A1, a specific inhibitor of V-type H(+) translocating ATPase (V-ATPase) or amiloride, an inhibitor of Na+/H+ exchange. Lidocaine slowed pHi recovery in a dose-dependent manner. Pretreatment (1 h) with 2.5 mM lidocaine abolished Na+/H+ exchange and reduced the V-ATPase-mediated component of pHi recovery. Lidocaine also significantly depressed the superoxide production induced by phorbol ester. TNF-alpha release induced by endotoxin was not affected significantly by the local anesthetic. Macrophage viability (trypan blue exclusion) and cellular ATP concentration were unaffected. These results indicate that lidocaine inhibits pHi regulatory mechanisms in alveolar m phi. This disruption of pHi regulation could contribute to inhibitory actions of lidocaine on m phi effector functions. PMID- 9330245 TI - Nontraumatic bilateral diaphragmatic myopathy: an unusual disturbance. AB - Bilateral diaphragmatic paralysis (BDP) is an uncommon entity among neuromuscular diseases. We present a young female with no triggering factors who developed progressive dyspnea with respiratory failure. Further studies led to the diagnosis of BDP. PMID- 9330246 TI - Modulation of airway responsiveness to acetylcholine by nitric oxide in a rabbit model. AB - Nitric oxide (NO) is an important mediator in the regulation of bronchial muscle tone and airway responsiveness. We investigated the influence of exogenous NO on airway responsiveness to acetylcholine aerosols (ACH) in normal and in hyperresponsive rabbits. White New Zealand rabbits were anesthetized, intubated, and breathed room air spontaneously. Responses of respiratory parameters in ACH challenge tests were measured. In group A the influence of NO on ACH infusion induced airway constriction was measured. Airway responses to aerosols from 0.25 to 8.0% ACH solutions in saline were measured with 150 and 300 ppm NO inhalation (groups B and C) and compared with the same animals' responses without NO. Moreover, we examined the influence of NO synthase inhibition on airway responsiveness (group D) and the modulatory effect of NO in hyperresponsive animals (group E). 300 ppm NO inhalation significantly decreased the bronchoconstrictor response to intravenously administered ACH (group A). However, the baseline value of dynamic elastance (Edyn) was only marginally lower under the influence of 300 ppm NO. During inhalation of 150 or 300 ppm NO, responses to nebulized 2.0% and less ACH solutions remained nearly unaltered. Responses to aerosols of 4.0 and 8.0% diminished significantly (groups B and C). Following 40 min of aerosolized N-nitro-L-arginine-methyl ester (L-NAME) solution (a NO synthase inhibitor, 1.2 mM) inhalation, the response of Edyn to ACH increased significantly in group D. In group E, animals inhaled 500 mg/m3 ammonium persulfate (APS), an oxidant with various industrial applications, after the first ACH challenge test (0.2, 1.0, and 2.0% ACH). After 2 h of APS exposure, the ACH-induced broncho constriction was increased significantly in the challenge test. After another 2 h of APS inhalation, the airway responsiveness to ACH was tested under the influence of 300 ppm NO. NO significantly decreased the response to ACH to almost the same level as before APS exposure. The results indicate that responses to high ACH concentrations as well as an APS-induced increase in ACH responsiveness were effectively reduced by high concentrations of inhaled NO. PMID- 9330247 TI - Detection of interstitial pneumonitis in patients with rheumatoid arthritis by measuring circulating levels of KL-6, a human MUC1 mucin. AB - In rheumatoid arthritis (RA), interstitial-pneumonitis is one of the major extraarticular complications that worsens a patient's prognosis. KL-6, a human MUC1 mucin, has been reported to be a sensitive serum marker for activity of interstitial pneumonitis. We investigated the clinical significance of serum KL-6 level in patients with RA. Serum levels of KL-6 and RA-associated inflammatory markers were evaluated in 177 RA patients. The diagnosis of active interstitial pneumonitis was made by clinical symptoms, pulmonary function tests, chest X-ray film, and high resolution CT. Serum KL-6 was increased in 8 of 9 (88.9%) RA patients with active interstitial pneumonitis but in only 1 of 168 (0.6%) RA patients without active interstitial pneumonitis. No significant correlation was found between KL-6 level and conventional clinical parameters. In RA, abnormal elevation of serum KL-6 strongly indicates the complication of active interstitial pneumonitis. PMID- 9330248 TI - Lymphocyte subsets in peripheral blood and smoking habits. AB - The investigation of peripheral blood lymphocyte (PBL) subpopulations is of interest in a wide variety of inflammatory diseases. Since the number of circulating lymphocytes has been shown to be affected by smoking habits, it seems useful to know how PBL subpopulations are influenced. We therefore determined percentages and absolute numbers of a wide range of PBL subpopulations in smokers (n = 14) and nonsmokers (n = 14). PBLs were obtained from healthy volunteers and analyzed by flow cytometry using antibodies for the detection of CD3, CD4, CD8, CD19, CD56, CD57, CD45RO, CD45RA, alpha/beta and gamma/delta T cell receptor epitopes. With the exception of CD3+ cells, no differences between smokers and nonsmokers were found regarding percentages of PBL subpopulations. Smokers were found to have higher absolute numbers of PBLs in the following subpopulations compared with nonsmokers: CD3+, CD4+, CD3+ alpha/beta +, CD45RO+/CD4+, and CD45RA+/CD4+. Cytotoxic lymphocytes, natural killer cells, and B cells did not differ in number between smokers and nonsmokers. There was likewise no difference in the number of the CD8+ alpha/beta + and all cells bearing the gamma/delta T cell receptor. Smoking increased the number of T cells and mainly CD4+ PBLs. The smoking habits of healthy control groups should therefore be taken into account when comparing lymphocyte subpopulations in different diseases. PMID- 9330249 TI - Lung tissue resistance measured in saline-filled guinea pig lungs by micropuncture. AB - Lung tissue resistance (Rti) measured in air-filled guinea pig lungs by the alveolar capsule technique was a large part of total lung resistance (Rl), and we wondered whether similar results applied to saline-filled lungs. We used the micropuncture method to measure alveolar pressure (Palv) in saline-filled lungs of 21 guinea pigs. Palv and airway opening pressure (Pao) were measured before and after a sudden interruption of flow during an inflation or deflation maneuver. On stopping flow, there was an immediate large change in Pao followed by a smaller slower change in Pao. Palv was nearly constant immediately after flow interruption but followed the slower change in Pao. The initial change in Pao on flow interruption was interpreted as the resistive pressure loss in the airways. The small change in Pao and Palv was interpreted as the pressure loss caused by tissue stress adaptation. Airway resistance (R(aw)) and Rti were obtained by dividing the pressure losses by the flow before the interruption. Rl was the sum of R(aw) and Rti. The calcium blocker nifedipine reduced both R(aw) and Rti and abolished the difference in Rti between inflation and deflation. Values of Rti were 10-29% of Rl. However, with correction for viscosity, Rti predicted in air-filled lungs would dominate Rl. PMID- 9330251 TI - A circadian rhythm of locomotor activity in newly emerged Ceratophyllus sciurorum. AB - Circadian rhythm in newly emerged individuals of the Red Squirrel (Scuirus vulgaris) flea C.s.sciurorum was studied in a constant environment, using an insect activity monitor. Trials were run over 7 days using two start times (08.00 and 17.00 hours). The results show that, regardless of start time, the fleas display a 24 h activity rhythm. The presence of a rhythm under constant conditions gives a strong indication that C.s.sciurorum has a self-sustaining clock which is started by disturbance and is most likely to be linked to host activity patterns. PMID- 9330250 TI - The spatial and seasonal distribution of African horse sickness and its potential Culicoides vectors in Morocco. AB - African horse sickness (AHS) is a vector-borne, infectious disease of equines that is caused by African horse sickness virus (AHSV). The only proven field vector is the biting midge Culicoides imicola, although C. obsoletus and C. pulicaris are suspected vectors. There was a recent epizootic of AHS in Iberia (1987-90) and Morocco (1989-91). In 1994-45 a total of 3887 light trap samples were taken from twenty-two sites distributed over most of Morocco. Culicoides imicola was found to be very widely dispersed, with the greatest catches in the low-lying northwestern areas (between Tangier and Rabat) and at Marrakech. Culicoides imicola was absent at one site only, near Settat. Culicoides imicola was found at altitudes ranging from 4 to 1275 m and in climatic conditions ranging from subhumid to saharan. In general, the catch of C.imicola peaked in late summer and autumn, with a smaller peak in spring. In areas where the insect appears most abundant at least one adult C.imicola per night may be caught in a light trap at all times of year, thus providing a possible means of viral overwintering. Culicoides obsoletus and C.pulicaris are also widely distributed in Morocco but trap catches were much lower than for C.imicola. Peak catches occurred in spring, and late summer and autumn. Other frequently caught species were C.circumscriptus, C.newsteadi, C.puncticollis and members of the odibilis subgenus. In general, the findings for C.imicola correspond well with the distribution of disease outbreaks during the epizootic. Although disease outbreaks were widespread in the country, the greatest number of reported cases was in the northwest (1989-90); in 1991 there were also significant numbers in Marrakech province. No cases were reported in a large area to the west of the Atlas mountains (including Settat) despite the presence of a large equine population. It is likely that during the epizootic the virus overwintered in the northwest (1989) and in Marrakech province (1990). Disease outbreaks occurred from July to December, with a peak from September to November. An unexplained phenomenon is the large number of reported cases of AHS in mules in Chefchaouen province in 1990, despite the apparent low abundance of C.imicola at a site at Chefchaouen. It is argued that C.obsoletus and C.pulicaris were probably of little significance to the epidemiology of AHS in Morocco in 1989-91. PMID- 9330252 TI - Mouthparts, antennae and genitalia of intersex Culicoides stellifer parasitized by mermithid nematodes. AB - A 12-month study of Culicoides in Bulloch County, Georgia, U.S.A., revealed that twelve of 23,859 specimens (0.05%) of C.stellifer were intersexes as a result of being parasitized by mermithid nematodes. Of these, eleven had male genitalia and female type antennae and one had female genitalia with male type antennae. PMID- 9330253 TI - Univariate analysis of tsetse habitat in the common fly belt of southern Africa using climate and remotely sensed vegetation data. AB - Tsetse are vectors of trypanosomes that cause diseases both in humans and livestock. Traditional tsetse surveys, using sampling methods such as Epsilon traps and black screen fly rounds, are often logistically difficult, costly and time-consuming. The distribution of tsetse, as revealed by such survey methods, is strongly influenced by environmental conditions, such as climate and vegetation cover, which may be readily mapped using satellite data. These data may be used to make predictions of the probable distribution of tsetse in unsurveyed areas by determining the environmental characteristics of areas of tsetse presence and absence in surveyed areas. The same methods may also be used to characterize differences between tsetse species and subspecies. In this paper we analyse the distribution of Glossina morsitans centralis, Glossina morsitans morsitans and Glossina pallidipes in southern Africa with respect to single environmental variables. For G.m.centralis the best predictions were made using the average NDVI (75% correct predictions; range > 0.37) and the average of the maximum temperature (70% correct predictions; 27.0-29.2 degrees C). For G.m.morsitans the best prediction was given by the maximum of the minimum temperature (84% correct predictions; range > 18.8 degrees C), and for G.pallidipes, also by the maximum of the minimum temperature (86% correct predictions; range > 19.6 degrees C). The following paper compares a range of multivariate techniques for making predictions about the distribution of these species in the same region. PMID- 9330254 TI - Mapping tsetse habitat suitability in the common fly belt of southern Africa using multivariate analysis of climate and remotely sensed vegetation data. AB - The distribution of Glossina morsitans centralis, Glossina morsitans morsitans and Glossina pallidipes are described in part of southern Africa, using a range of multivariate techniques applied to climate and remotely sensed vegetation data. Linear discriminant analysis is limited in its predictive power by the assumption of common covariances in the classes within multivariate environment space. Maximum likelihood classification is one of a variety of alternative methods that do not have this constraint, and produce a better prediction, particularly when a priori probabilities of presence and absence are taken into account. The best predictions are obtained when the habitat is subdivided, prior to classification, on the basis of a bimodality detected on the third component axis of a principal component analysis. The results of the predictions were good, particularly for G.m.centralis and G.m.morsitans, which gave overall correct predictions of 92.8% and 85.1%, with a Kappa index of agreement between the prediction and the training data of 0.7305 and 0.641 respectively. For G.pallidipes, 91.7% of predictions were correct but the value of Kappa was only 0.549. Very clear differences are demonstrated between the habitats of the two subspecies G.m.centralis and G.m.morsitans. PMID- 9330255 TI - Comparison of the susceptibility of different Glossina species to simple and mixed infections with Trypanosoma (Nannomonas) congolense savannah and riverine forest types. AB - Teneral Glossina morsitans mositans, G.m.submorsitans, G.palpalis gambiensis and G.tachinoides were allowed to feed on rabbits infected with Trypanosoma congolense savannah type or on mice infected with T.congolense riverine-forest type. The four tsetse species and subspecies were also infected simultaneously in vitro on the blood of mice infected with the two clones of T.congolense via a silicone membrane. The infected tsetse were maintained on rabbits and from the day 25 after the infective feed, the surviving tsetse were dissected in order to determine the infection rates. Results showed higher mature infection rates in morsitans-group tsetse flies than in palpalis-group tsetse flies when infected with the savannah type of T.congolense. In contrast, infection rates with the riverine-forest type of T.congolense were lower, and fewer flies showed full development cycle. The intrinsec vectorial capacity of G.m.submorsitans for the two T.congolense types was the highest, whereas the intrinsic vectorial capacity of G.p.gambiensis for the Savannah type and G.m.morsitans for the riverine-forest type were the lowest. Among all tsetse which were infected simultaneously with the two types of T.congolense, the polymerase chain reaction detected only five flies which had both trypanosome taxa in the midgut and the proboscis. All the other infections were attributable to the savannah type. The differences in the gut of different Glossina species and subspecies allowing these two sub-groups of T.congolense to survive better and undergo the complete developmental cycle more readily in some species than other are discussed. PMID- 9330256 TI - Blowfly species composition in sheep myiasis in Scotland. AB - Samples of dipteran larvae were collected from live sheep throughout Scotland, reared in the laboratory, and identified once adult flies emerged. Lucilia sericata was found in 77% of samples, and other species in 49%. The most common alternative species were L.caesar, which occurred in 31% of samples, and Protophormia terraenovae, which occurred in 18%. Three other calliphorid species, Calliphora vomitoria, C.vicina and L.illustris, and the muscid Muscina pabulorum were also found. The proportion of samples containing alternative species was significantly lower in eastern Scotland than in western Scotland. Significantly higher proportions of samples containing alternative species were collected at altitudes of 200m and above; from sheep of hill breeds; from rough grazing conditions and moorland; in the absence of trees; and in the presence of bracken. PMID- 9330257 TI - The effects of hair density of beef cattle on Haematobia irritans horn fly populations. AB - We show the relationships that exist between the amount of hair and quantity of sebum on cattle skin and the population density of the horn fly, Haematobia irritans. Brahman and Chianina steers had means of 2390 and 1587 hairs per cm2, respectively, significantly more than the mean number of hairs on Angus, Brahman x Angus Crossbred, Charolais, and Red Poll steers. The Chianina steers had > 30% more sebum present on their skin and hair (0.58g/929 cm2) than the Angus, Charolais, and Red Poll steers at the Beef Cattle Research Station Savoy, Arkansas. The Brahman steers had a significantly greater amount of sebum present on the skin (1.51 g/929 cm2) than the Crossbred and purebred Angus steers (0.55 and 0.25 g/929 cm2, respectively) at the South Central Family Farms Research Centre Booneville, Arkansas. The Brahman and Chianina steers had means of 61.9 and 17.0 horn flies per steer, respectively, during the fly season, whereas the Angus, Crossbred, Charolais and Red Poll steers had fly season means that ranged from 76.9 to 265.8 flies per steer. Regression analysis showed that an increase of 100 hairs per cm2, was associated with a reduction of 11 horn flies in the Angus II, 5 in Angus I, 20 in Charolais, 37 in Red Poll, and 0.4 in Chianina steers at the Savoy Station and a reduction of 6.6 horn flies for the Angus, Brahman, and Crossbred steers at the Booneville Centre. Regardless of cattle breed, an increase of 1.0 g of sebum per 929 cm2 output by the steer was associated with 478.5 additional hairs per cm2 on the animal. Each increase of 0.25 g of sebum per 929 cm2 resulted in a decrease of 9.2 horn flies per steer. We conclude that some of the factors responsible for fly-resistance in cattle are hair density and the corresponding amount of sebum present on cattle skin and hair. PMID- 9330258 TI - Effects of Plasmodium yoelii nigeriensis infection on Anopheles stephensi egg development and resorption. AB - It has been shown previously that infection with Plasmodium yoelii nigeriensis reduces the number of eggs produced by female Anopheles stephensi. Here we examine the mechanism underlying fecundity reduction. Ovaries from infected and uninfected (control) female mosquitoes were examined 12, 24 or 36 h after blood feeding during the first gonotrophic cycle (replicated) or the second gonotrophic cycle (unreplicated). Follicular development was assessed according to Christophers' stages and the proportions of developing and resorbing follicles per ovary were determined. Resorption of some follicles commenced within 12 h of blood-feeding, affecting significantly more follicles in the infected females: 1.1% v. 3.2%. The difference was greatest 36 h after blood-feeding: 25% reduction (10 v. 35%) in the first cycle; 16% reduction (9 v. 25%) in the second gonotrophic cycle. The mean speed of oogenesis was also found to be significantly retarded in infected mosquitoes. During the second gonotrophic cycle, for example, only 92-94% of follicles reached stage III by 24 h and stage IV by 36 h in infected females, whereas all the developing follicles of uninfected females reached these stages more or less synchronously in the time specified. PMID- 9330259 TI - Immunology of interactions between ticks and hosts. AB - Infestation with ixodid tick stimulates the immune regulatory and effector pathways of the hosts involving antigen presenting cells, T-lymphocytes, B lymphocytes, basophils, mast cells, eosinophils and a variety of bioactive molecules like cytokines, antibodies and complement. Tick-mediated immunosuppression has been investigated using cells derived from infested animals and by exposing cells from uninfected animals to tick salivary gland molecules. Tick-induced suppression of host immune defences is characterized by reduced ability of lymphocytes from infested animals to proliferate in vitro in the presence of concanavalin A (Con A), diminished primary antibody responses to T cell dependent antigen, and decreased elaboration of macrophage (IL-1 and TNF alpha) and Th1-lymphocyte cytokines (IFN-gamma), whereas Th2 cytokines production (IL-4, IL-5 and IL-10) is enhanced. It is known that IL-10 inhibits Th1 cell development and also reduces the in vitro T-lymphocyte proliferative response to Con A stimulation. Proteins which inhibited T-lymphocyte in vitro responsiveness to Con A were also isolated from tick salivary glands. PMID- 9330260 TI - Tick saliva: recent advances and implications for vector competence. AB - Secretions of the tick salivary glands are essential to the successful completion of the prolonged feeding of these ectoparasites as well as the conduit by which most tick-borne pathogens are transmitted to the host. In ixodid ticks the salivary glands are the organs of osmoregulation, and excess water from the bloodmeal is returned via saliva into the host. Host blood must continue to flow into the feeding lesion as well as remain fluid in the tick mouthparts and gut. The host's haemostatic mechanisms are thwarted by various anti-platelet aggregatory, anticoagulatory and anti-vasoconstrictory factors in tick saliva. Saliva components suppress the immune and inflammatory response of the host permitting the ticks to remain on the host for an extended period of time and, adventitiously, enhancing the transmission and establishment of tick-borne pathogens. Over the years much work has been done on the numerous enzyme and pharmacological activities found in the tick saliva. The present article reviews the most recent work on salivary gland secretions with special emphasis on how they favour pathogen transmission. PMID- 9330261 TI - Control of Trypanosoma brucei brucei infections in tsetse, Glossina morsitans. AB - Numbers of immature Trypanosoma brucei brucei within a tsetse midgut remain remarkably constant after establishment throughout the course of an infection, irrespective of whether the infection eventually matures. These results suggest a system of self regulation of the parasite population in the insect gut based on a form of programmed cell death which would carry advantages for both the parasite and the vector. PMID- 9330262 TI - Interactions of human malaria parasites, Plasmodium vivax and P.falciparum, with the midgut of Anopheles mosquitoes. AB - Present understanding of the development of sexual stages of the human malaria parasites Plasmodium vivax and P.falciparum in the Anopheles vector is reviewed, with particular reference to the role of the mosquito midgut in establishing an infection. The sexual stages of the parasite, the gametocytes, are formed in human erythrocytes. The changes in temperature and pH encountered by the gametocyte induce gametogenesis in the lumen of the midgut. Macromolecules derived from mosquito tissue and second messenger pathways regulate events leading to fertilization. In An.tessellatus the movement of the ookinete from the lumen to the midgut epithelium is linked to the release of trypsin in the midgut and the peritrophic matrix is not a firm barrier to this movement. The passage of the P.vivax ookinete through the peritrophic matrix may take place before the latter is fully formed. The late ookinete development in P.falciparum requires chitinase to facilitate penetration of the peritrophic matrix. Recognition sites for the ookinetes are present on the midgut epithelial cells. N-acetyl glucosamine residues in the oligosaccharide side chains of An.tessellatus midgut glycoproteins and peritrophic matrix proteoglycan may function as recognition sites for P.vivax and P.falciparum ookinetes. It is possible that ookinetes penetrating epithelial cells produce stress in the vector. Mosquito molecules may be involved in oocyst development in the basal lamina, and encapsulation of the parasite occurs in vectors that are refractory to the parasite. Detailed knowledge of vector-parasite interactions, particularly in the midgut and the identification of critical mosquito molecules offers prospects for manipulating the vector for the control of malaria. PMID- 9330263 TI - Odour attractants for tsetse: Glossina austeni, G.brevipalpis and G.swynnertoni. PMID- 9330264 TI - Therapeutic efficacy of linalool for the topical treatment of parasitic otitis caused by Psoroptes cuniculi in the rabbit and in the goat. PMID- 9330265 TI - Effect of all-trans-retinoic acid alone or in combination with chemotherapy in newly diagnosed acute promyelocytic leukaemia. AB - Between February 1992 and November 1996 we treated 30 newly diagnosed acute promyelocytic leukaemia (APL) patients either with oral all-trans-retinoic acid (ATRA) alone (45 mg m-2) or with a simultaneous combination of ATRA (45 mg m-2), daunorubicin (DNR, 50 mg/m-2 for 3 days) and cytosine arabinoside (ARA-C, 200 mg m-2 for 7 days). There were 15 patients in each group. Patients with a white blood cell count < 5 x 10(9)/l at diagnosis received only ATRA as an induction therapy. Patients with initial white blood cell count > 5 x 10(9)/l received a combination of ATRA, DNR and ARA-C as an induction therapy. Within the first 20 days of induction, there were two early deaths in the group of patients receiving only ATRA, and six early deaths in the group of patients treated with a combination of ATRA and chemotherapy. Ten out of 13 patients (76.9%) receiving ATRA only achieved complete remission (CR) whereas seven out of nine patients (77.8%) receiving ATRA with chemotherapy achieved CR. Initial median peripheral white blood cell counts were significantly lower in the group of patients treated with ATRA alone (2.3 x 10(9)/l) than in the group of patients receiving ATRA and chemotherapy (14.0 x 10(9)/l). Morphological evidence of differentiation was noted in all patients entering CR. Patients in both groups who achieved CR received one course of standard '3 + 7' chemotherapy (DNR 45 mg m-2, 1-3 days, ARA-C 200 mg m-2, 1-7 days) followed by two courses of standard '2 + 5' chemotherapy (DNR 50 mg m-2 1-2 days, ARA-C 200 mg m-2 1-5 days) as a consolidation therapy. Patients not achieving remission (three out of 13 in the ATRA group and two out of nine in ATRA+chemotherapy group) did not respond to salvage chemotherapy and all died within 3 months of diagnosis. Only one out of 10 patients (10%) in CR, treated with ATRA is in relapse after 18 months. In patients treated with ATRA alone two out of 10 (20%) survived 58 months following diagnosis whereas in the ATRA+chemotherapy group one out of seven has already survived their 58th month since diagnosis. Four out of eight patients with an early death died of retinoic acid syndrome. Other toxicities due to ATRA were minimal (cheilitis, xerosis, dermatitis, diarrhoea, liver damage or pseudotumor cerebri). PMID- 9330266 TI - Systemic therapy of malignant melanoma. AB - The present status of medical treatment of malignant melanoma is briefly reviewed, both with regard to adjuvant therapy for individuals with high-risk melanoma and a high probability of harbouring subclinical micrometastases, as well as to therapy for established disseminated (macrometastatic) disease. At present, disseminated, macrometastatic melanoma is incurable in the majority of cases. Single agent chemotherapy has modest effects and results in disease remission in a minority of patients, usually of short duration, Combination chemotherapy, or the combination of chemotherapeutic drugs and cytokines, results in increased response rates and occasionally remissions of prolonged duration. So far, no regimen has demonstrated improved survival compared to single agent therapy in disseminated melanoma. New insights into the mechanisms of resistance to chemotherapeutic drugs may lead to development of predictive tests that can identify individuals with tumors sensitive to a specific agent, as well as to the development of strategies to circumvent drug resistance. It has recently been shown that adjuvant therapy of high-risk melanoma with large doses of interferon alpha 2b significantly prolongs relapse-free and overall survival, at the price of considerable toxicity. Ongoing studies aim to define the optimum dose and duration of adjuvant interferon therapy. Recent advances in molecular biology and immunology may lead to the development of new treatment modalities, such as improved vaccines and other biologic therapies, which may benefit patients with malignant melanoma. PMID- 9330267 TI - Targeting signal transduction for disease therapy. AB - With the advance in the molecular understanding of cancers and proliferative disorders new approaches to managing these diseases may become feasible. It has been recognized that a key feature of these diseases is the pathological alteration in the molecular machineries of signalling pathways. This recognition which began to emerge in the early 1980s induced us to explore the possibility of targetting the aberrant signalling pathways for disease therapy. I now present evidence for the validity of the approach. PMID- 9330268 TI - Use of granulocyte-macrophage colony stimulating factor in the treatment of prolonged haematopoietic dysfunction after chemotherapy alone or chemotherapy plus bone marrow transplantation. AB - This study evaluates the use of granulocyte-macrophage colony stimulating factor (GM-CSF) in patients with prolonged haematopoietic dysfunction (> 21 days) after using chemotherapy to treat cancer. One hundred and seven patients were identified who had a leucocyte count below 1000 cells/mm3 more than 21 days after start of chemotherapy (81 patients) or after bone marrow transplantation (BMT)(26 patients). There were 66 males and 40 females ranging in age from 4.5 to 82 years. The duration of aplasia was 48 +/- 43 days in the chemotherapy alone group, and 79 +/- 57 days in the post BMT group. Over 80% of the patients had haematologic malignancies and 70% had an infection prior to the start of the cytokine. Patients received 5 micrograms GM-CSF/kg1 body weight daily i.v. or s.c. for 14 +/- 11 days in the chemotherapy group and 20 +/- 26 days in the BMT group. Sixty percent of chemotherapy patients and 58% of BMT patients had a haematological response to treatment (leucocyte count > 2000 cells/mm3. Median times to haematologic recovery were 7 days in the chemotherapy group and 10 days in the BMT group. There was a significant reduction in the number of infections (73% to 28% in the chemotherapy group). Clinical responses in the two groups were 55% and 50%, respectively. No severe, drug-related adverse events were reported and no evidence of stimulation of malignant clones was observed. It is concluded that GM-CSF is effective and well tolerated in patients with prolonged bone marrow dysfunction after chemotherapy or BMT. Although results from an open-label trial must be viewed with caution, this observation confirms the value and safety of GM-CSF therapy in patients with this severe, and often fatal, condition. PMID- 9330269 TI - Haemopoietic cell composition of human fetal liver, spleen and thymus. AB - The haemopoietic cell composition of fetal liver, spleen and thymus was studied in human aborted fetuses of 12-22 weeks gestation. Erythroid cells were present primarily in liver and to a lesser extent in spleen. Orthochromatic normoblasts were the predominant erythroid component. Lymphoid cells were seen primarily in the thymus and to a lesser extent in the spleen. There were few granulocytic and megakaryocytic cells in all these organs. A few primitive haemopoietic cells (Haemocytoblasts) were identified only in liver. These observations indicate that during mid-fetal life (12-22 weeks) the liver contributes primarily to erythropoiesis, the thymus to lymphopoiesis, the spleen to both, and that there is a lack of granulopoiesis and megakaryopoiesis at these sites. PMID- 9330271 TI - Primary neuroendocrine tumor of the breast. AB - Adult neuroendocrine tumors may present with a wide range of clinical symptoms that share specific ultrastructural and biochemical features. A 63-year-old post menopausal female patient was admitted to the hospital with a mass in her right breast and the diagnosis was primary neuroendocrine tumor of the breast. Although neuroendocrine tumors can originate in various parts of the body and breast carcinoma with neuroendocrine differentiation has been described, primary neuroendocrine tumor of the breast is very unusual. This case is now presented and the current literature is reviewed. PMID- 9330272 TI - Generation and characterization of cellular retinoic acid-binding proteins from Escherichia coli expression systems. PMID- 9330273 TI - Generating and characterizing retinoid receptors from Escherichia coli and insect cell expression systems. PMID- 9330274 TI - Expression and characterization of retinoid receptors in yeast. PMID- 9330270 TI - Current perspectives in gliomas. AB - The annual incidence of primary central nervous system tumors, including gliomas, is increasing, however, the prognosis of these tumors remains poor with a median survival of only 5 years. The imaging of tumors by computerised tomography, magnetic resonance imaging and newer methods such as positron emission tomography and proton magnetic resonance spectroscopy (1H-MRS) is increasing our knowledge of tumor biology and extent of the disease. Advances within the field of neurosurgery have improved operative procedures reducing mortality and morbidity. Furthermore, radiotherapy planning, tumor targeting and repositioning for treatment have all improved initial tumor management. The role of adjuvant chemotherapy remains controversial. Chemotherapy for advanced and recurrent disease has been extensively investigated, and although improvements in quality of life have been recorded, no prolongation of survival has been documented. With new discoveries and increasing knowledge of the physiology and molecular biology of these tumors the potential for targeting therapy at a genetic level is becoming increasingly promising. This review provides an overview of these current perspectives in glioma management. PMID- 9330275 TI - Use of in situ hybridization techniques to study embryonic expression of retinoid receptors and binding proteins. PMID- 9330276 TI - Use of quantitative polymerase chain reaction to study retinoid receptor expression. PMID- 9330277 TI - Use of quantitative polymerase chain reaction to study cellular retinoic acid binding protein-II mRNA expression in human skin. PMID- 9330278 TI - Use of transgenic mice to study activation of retinoic acid-responsive promoters. PMID- 9330279 TI - Use of transgenic mice to eliminate retinoic acid receptor function in specific tissues. PMID- 9330280 TI - Use of reporter cells to study endogenous retinoid sources in embryonic tissues. PMID- 9330281 TI - Preparation of radiolabeled 9-cis- and all-trans-retinoids. PMID- 9330282 TI - Identification and quantification of retinoic acid and other metabolites from beta-carotene excentric cleavage in human intestine in vitro and ferret intestine in vivo. PMID- 9330283 TI - Assessing metabolism of beta-[13C]carotene using high-precision isotope ratio mass spectrometry. PMID- 9330284 TI - Atmospheric pressure chemical ionization and electron capture negative chemical ionization mass spectrometry in studying beta-carotene conversion to retinol in humans. PMID- 9330285 TI - Synthesis of [3 alpha-3H]vitamin D3 and 1 alpha,25-dihydroxy[1 beta-3H]vitamin D3. PMID- 9330286 TI - Assay of 1,25-dihydroxyvitamin D3 from serum samples: use of receptor-binding or enzyme-coupled reporter analysis. PMID- 9330287 TI - Quantitation of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D by radioimmunoassay using radioiodinated tracers. PMID- 9330288 TI - Assay of vitamin D derivatives and purification of vitamin D hydroxylases. PMID- 9330289 TI - Assay of 25-hydroxyvitamin D 1 alpha-hydroxylase and 24-hydroxylase. PMID- 9330290 TI - Molecular cloning of vitamin D3 hydroxylases. PMID- 9330291 TI - Role of 1 alpha,25-dihydroxyvitamin D3 in osteoclast differentiation and function. PMID- 9330292 TI - Assay of direct effect of 1,25-dihydroxyvitamin D3 on calcium ion influx into cultured osteoblasts. PMID- 9330293 TI - Vitamin E status and immune function. AB - Evidence from animal and human studies indicates that vitamin E plays an important role in the maintenance of the immune system. Even a marginal vitamin E deficiency impairs the immune response, while supplementation with higher than recommended dietary levels of vitamin E enhances humoral and cell-mediated immunity. The current RDA level of vitamin E prevents clinical deficiency syndrome but in some situations, especially in older subjects or in a disease state, fails to maintain optimal host defense. The immunological parameters reviewed are all sensitive to changes in the availability of vitamin E and, therefore, may reflect the vitamin E status of a given individual more accurately than conventional methods. PMID- 9330294 TI - Inhibition of platelet adhesion as functional test for vitamin E status. PMID- 9330295 TI - Inhibition of plasma cholesterol ester hydroperoxide and phosphatidylcholine hydroperoxide formation as measures of antioxidant status. PMID- 9330296 TI - alpha-tocopherol-binding proteins: purification and characterization. PMID- 9330297 TI - alpha-Carboxyethyl-6-hydroxychroman as urinary metabolite of vitamin E. PMID- 9330298 TI - Purification of vitamin K-dependent carboxylase from cultured cells. PMID- 9330299 TI - Purification of native bovine carboxylase and expression and purification of recombinant bovine carboxylase. PMID- 9330300 TI - Purification of gamma-glutamyl carboxylase from bovine liver. PMID- 9330301 TI - Assay of vitamin K-dependent carboxylase activity in hepatic and extrahepatic tissues. PMID- 9330302 TI - Expression of human anticoagulation protein C and gamma-carboxyglutamic acid mutants in mammalian cell cultures. PMID- 9330303 TI - Determination of site-specific gamma-carboxyglutamic acid formation by vitamin K dependent carboxylase utilizing De-gamma-carboxy bone Gla protein as substrate. PMID- 9330304 TI - Purification of warfarin-sensitive vitamin K epoxide reductase. PMID- 9330305 TI - Determination of vitamin K compounds in plasma or serum by high-performance liquid chromatography using postcolumn chemical reduction and fluorimetric detection. PMID- 9330306 TI - Assay of phylloquinone in plasma by high-performance liquid chromatography with electrochemical detection. PMID- 9330307 TI - Assay of menaquinones in plasma utilizing dual-electrode electrochemical detection. PMID- 9330308 TI - Assay of phylloquinone and menaquinones in human liver. PMID- 9330309 TI - Determination of phylloquinone in foods by high-performance liquid chromatography. PMID- 9330310 TI - Assay of menaquinones in bacterial cultures, stool samples, and intestinal contents. PMID- 9330312 TI - Purification and identification of human and mouse granulocyte chemotactic protein-2 isoforms. PMID- 9330311 TI - Expression, purification, and characterization of Escherichia coli-derived recombinant human melanoma growth stimulating activity. PMID- 9330313 TI - Synthesis of chemokines by native chemical ligation. PMID- 9330314 TI - Molecular approaches to structure-function analysis of interleukin-8. PMID- 9330315 TI - Alanine scan mutagenesis of chemokines. PMID- 9330316 TI - Biological assays for C-X-C chemokines. PMID- 9330317 TI - Characterization of quaternary structure of interleukin-8 and functional implications. PMID- 9330318 TI - Isolation of human monocyte chemotactic proteins and study of their producer and responder cells by immunotests and bioassays. PMID- 9330319 TI - Assays for macrophage inflammatory proteins. PMID- 9330320 TI - Gene expression of RANTES. PMID- 9330321 TI - Expression of chemokine RANTES and production of monoclonal antibodies. AB - RANTES was first identified as a cDNA in a search for genes expressed late (3-5 days) after T-cell activation. Definition of RANTES function depended on the generation of protein. This chapter describes the various techniques used to make recombinant RANTES protein, to test its activity, and to generate monoclonal antibodies to assess RANTES protein cell distribution. PMID- 9330322 TI - Biological responses to C-C chemokines. PMID- 9330323 TI - Gene targeting strategies to study chemokine function in vivo. PMID- 9330324 TI - Lymphotactin: a new class of chemokine. PMID- 9330325 TI - Isolation and purification of neutrophil-activating peptide-4: a chemokine missing two cysteines. PMID- 9330326 TI - Chemical synthesis, purification, and folding of C-X-C and C-C chemokines. PMID- 9330327 TI - Identification of inflammatory mediators by screening for glucocorticoid attenuated response genes. AB - We describe an approach for identifying novel inflammatory mediators, based on screening for immediate early/primary response genes whose induction by an inflammatory stimulus is attenuated by glucocorticoids. This procedure can be applied to a wide range of cell types and tissues, using a variety of inducers. In an initial test of this idea, we identified cDNAs for 12 LPS-induced, glucocorticoid-attenuated response genes (GARGs) by differential hybridization screening of a lambda phage cDNA library from murine 3T3 fibroblasts. Seven of the GARGs were known genes, including the chemokines JE/MCP-1, fic/MARC/MCP-3, and crg2/IP-10. One of the novel cDNAs was a new C-X-C chemokine that we designated LIX, for LPS-induced C-X-C chemokine. Because the 12 GARG cDNAs were identified in a single screening of only 15,000 phage, and four were found as single isolates, these results suggest that there are many GARGs not yet described. Furthermore, six of the seven known GARGs encode proteins that modulate intercellular communication. These results support our hypothesis that GARGs predominantly encode products that function in paracrine cell communication. Here we provide an overview of the GARG strategy and the differential hybridization procedures used in our initial screening. A variety of other methods for identifying differentially expressed genes may be used in future searches for novel GARGs. PMID- 9330328 TI - Chemokine-induced human lymphocyte infiltration and engraftment in huPBL-SCID mice. PMID- 9330329 TI - High throughput screening for identification of RANTES chemokine expression inhibitors. PMID- 9330330 TI - Transgenic methods to study chemokine function in lung and central nervous system. PMID- 9330331 TI - Chemokines and chemokine receptors in model neurological pathologies: molecular and immunocytochemical approaches. PMID- 9330332 TI - Synthesis and evaluation of fluorescent chemokines labeled at the amino terminal. PMID- 9330333 TI - Solid-phase binding assay to study interaction of chemokines with glycosaminoglycans. PMID- 9330334 TI - Biological activity C-X-C and C-C chemokines on leukocyte subpopulations in human whole blood. AB - We have described an assay that monitors the activating effects of a variety of chemokines on leukocyte subsets in human whole blood. This procedure has the following advantages: (1) minimal manipulation of the cells, (2) maintenance of more physiological conditions, and (3) simultaneous monitoring of the responses of monocytes, neutrophils, and eosinophils. PMID- 9330345 TI - Introduction to the biliary tract, the gallbladder, and gallstones. AB - This paper serves to introduce a topical section of fifteen invited original research contributions dealing with normal and pathological development of the human biliary tract. This section also includes comparative anatomy of the gallbladder and the cystic duct as well as, the formation of gallstone. This series of reports have used advanced microscopic and ancillary techniques to study adaptative changes in gallbladder epithelial cell changes regarding permeability, renewal, mucous secretion as well as cholesterol uptake and nucleation. Several contributions deal with the blood and lymphatic drainage of the gallbladder. The gallbladder contractility is clarified by recent findings about its innervation, elegantly demonstrated and supported by complementary immunohistochemical and neurophysiological techniques. In vivo models for production of cholelithiasis in the ground squirrel and the Syrian hamster are introduced. Recent in vitro cellular and molecular models have substantially increased the understanding of biliary tract calculi formation. Finally, a survey and new data about progesterone gene regulation of both cholesterol metabolism and gallstone formation obtained in the Syrian hamster model are compared with cholelithogenesis in human. PMID- 9330346 TI - Microstructure and development of the normal and pathologic biliary tract in humans, including blood supply. AB - Microstructure and development of the normal biliary tract and the pathologies of several biliary tract diseases in humans are reviewed. The biliary tract, comprising the bile duct and peribiliary glands, is anatomically divided into the extrahepatic and intrahepatic biliary tree. The intrahepatic biliary tree is further divided into large bile ducts, corresponding to the right and left hepatic ducts and their first to third order branches, and into septal and interlobular bile ducts and bile ductules according to their size and location relative to the hepatic lobules and surrounding structures. The right and left hepatic ducts and the extrahepatic bile ducts are composed of dense fibrous duct walls lined by a layer of columnar biliary epithelium. The peribiliary glands, which may secrete mucinous and serous substances into the bile, are found along the extrahepatic and large intrahepatic bile ducts. They are divided in glands within and outside the duct wall. The former (intramural glands) drain directly into the lumen of the bile duct, while the latter (extramural glands) are composed of several lobules and drain into the ductal lumen via their own conduits. The biliary tract is supplied by a complex vasculature called the peribiliary vascular plexus. Afferent vessels of this plexus derive from hepatic arterial branches, and this plexus drains into the portal venous system or directly hepatic sinusoids. The development of the intrahepatic biliary tract is divided into three stages: the stage of the ductal plate, the stage of biliary cell migration into the mesenchyme, and the stage of bile duct formation in the portal tract. It remains unclear how the extrahepatic and intrahepatic biliary tract integrate. Along with these developmental changes in the biliary tract, the peribiliary glands and the vascular plexus also develop in a step-wise manner and their maturation is completed after birth. Pathologies of various biliary diseases are briefly reviewed noting their relevance to several histologic elements and the microenvironment of the biliary tract and the developmental anomalies of the biliary tract including ductal plate malformation. PMID- 9330347 TI - Comparative morphology of the gallbladder and biliary tract in vertebrates: variation in structure, homology in function and gallstones. AB - A review of investigations on the morphology of the gallbladder and biliary tract in fish, reptiles, amphibians, birds, and mammals was performed. Scanning electron microscopy, transmission electron microscopy, and light microscopy observations by the authors were also included. Variations in the presence or absence of a gallbladder, surface epithelium of the gallbladder, and differences in the morphology of the biliary tract in vertebrates were reported. Many differences were diet-related. Despite some dissimilarities observed, analogous functioning of the biliary system was accomplished by its various components, with the biliary ducts performing the function of the gallbladder when this organ was absent. In addition, the occurrence of peculiar parasitism and gallstones among some cases of vertebrates, including humans, was presented. PMID- 9330348 TI - Presence of brush cells in the mouse gallbladder. AB - The brush cells (BC) are the second most frequent cellular component of the epithelium of the mouse gallbladder. They have a topographical distribution, being present in large numbers toward the neck and in the fundic regions of the organ and are scattered in the body. Serial section studies demonstrate that BC have a characteristic shape consisting of a narrow apical portion, bulky body and basal cytoplasmic projections. BC are located obliquely among the principal cells. Scanning electron microscopy demonstrates that the microvilli forming the prominent brush border, after which the cell was named, have a triangular arrangement. Due to their size and stiffness, the microvilli of BC have more similarity with stereocilia of sensory cells than with conventional microvilli. Furthermore freeze-fracture replicas demonstrate that, like stereocilia, the P face of the microvilli plasma membrane of BC is smoother than the E face but several intramembranous particles form small aggregates on the microvillus tip of both P and E faces. Numerous intramembranous particles are scattered on the lateral plasma membrane. An unusual, spatially organized cytoskeleton characterizes the apical cytoplasm of BC. The use of the appropriate fixative reveals that it consists of bundles of actin filaments originating from the axis of the apical microvilli and stretching continuously up to the supranuclear region of the cell. Microtubuli, also assembled in bundles, flank in alternating manner the actin filaments over their whole course. Due to the strong parallel arrangement of both cytoskeletal structures, the apical cytoplasm of the BC assumes a typical stiffness, observable in both thin sections and freeze-fracture replicas. A variable number of vesicles of different size are aligned between the bundles of actin filaments and microtubuli; their shape is highly influenced by the fixative used. Intraluminal injection of horseradish peroxidase demonstrates that these vesicles are not resorptive as they are not filled by the tracer. The BC possess a large number of lateral microvilli. These, whether single or in pairs, are rigid cytoplasmic protrusions that leave the lateral surface of the cell in all directions and penetrate deeply into the cytoplasm of the adjacent principal cells. The bundle of actin filaments emanating from each lateral microvillus extends at different angles into the cytoplasm. A conspicuous amount of bundles of 10 nm filaments is intertwined around the nucleus and extends toward the desmosomes of the lateral plasma membrane and into the basal cellular body. Arguments are considered in support of the view that interactions between the plasma membrane with its differentiations on the one hand and the cytoskeleton elements on the other hand, play a key role in the function of BC as a receptor (sensory) cell. PMID- 9330349 TI - DNA synthesis, cell proliferation index in normal and abnormal gallbladder epithelium. AB - The observation of mitotic figures in the epithelium of the normal gallbladder is exceptional because cell renewal is occurring at a very slow rate. It is only after using 3H-thymidine and autoradiography to observe the cells in DNA synthesis that evidence of a significant epithelial cell replication has been provided. Because numerous mitotic figures and increased 3H-thymidine uptake have been observed after intraluminal introduction of foreign bodies or after ligation of the common bile duct in animals, mechanical distension has been supposed to represent an important trigger factor of cell proliferation in this hollow organ. An increased epithelial cell renewal was also observed in human gallbladders of patients with a complete obstruction of the common bile duct causing the distension. However, the absence of correlation between the degree of gallbladder distension and the proliferative response was suggesting that factors other than distension could be involved. In studies on experimental lithiasis cell proliferation appeared to be enhanced in the gallbladder epithelium of mice fed on a cholesterol-cholic acid-rich lithogenic diet. The fact that the increase in proliferative activity was preceding the formation of gallstones was another indication that factors other than mechanical stimulation by stretching or by the stones may stimulate cell renewal in this organ. Factors in the bile of animals receiving a lithogenic diet could be involved which might cause cellular death and, hence, a regenerative reaction. Direct mitogenic effect of an unknown factor in the bile of these animals is an alternative possibility. On the other hand the stimulating effect of postprandial hormones on gallbladder cell renewal suggested by the observation of a DNA synthesis peak after feeding has been established. Synthetic cholecystokinin analogues have been shown to increase the proliferative activity and to induce epithelial hyperplasia in this organ. In one recent study using fundusectomy to increase the serum gastrin levels, a significant proliferative stimulation in the gallbladder was also observed. In human gallbladder mitotic activity in gallbladders with gallstones in much higher than in the controls. No correlation between stone number, weight or volume and the proliferative activity was put in evidence, whereas cell renewal appeared to be more influenced by the composition of the stones than by their physical presence. Epithelial DNA synthesis activity was, namely, much higher in gallbladders with cholesterol stones than in those with pigment stones. Whether increased cell turnover and, hence, cellular shedding into the lumen could represent a nucleating factor for cholesterol stones is an attractive working hypothesis. Considering the very high frequency of gallstones in man and also the frequent association of gallbladder cancer and lithiasis, further studies on mitotic activity in this organ are required. In conclusion, data from animal experiments and in vitro studies on human gallbladders indicate that gallbladder epithelial cell proliferation may be influenced by several mechanical, chemical and hormonal factors. The list of these factors is still incomplete while their possible role in gallbladder disease is a fascinating exploration field for future research. PMID- 9330351 TI - Biochemical and morphological correlations in human gallbladder with reference to membrane permeability. AB - There is good evidence that gallbladder epithelium is permeable to a diverse range of molecules which move into the epithelial cell from the lumen or the basement membrane. Morphological investigations have shown both secretory mucous droplets, components of the endocytosis pathway together with evidence of a system allowing passage of molecules across the basement membrane. This indicates that the gallbladder epithelium may be influenced by molecules presented via the apical and basal membranes, complicating our understanding of gallbladder function, particularly in disease. Gallbladder disease increases the proteoglycan content of the basement membrane, but the implication of this in terms of permeability remains to be defined. Indeed, it remains unknown whether this precedes disease or is a manifestation of the disease process. The removal of water from hepatic bile by gallbladder involves two counter ion transport systems. Autoradiography shows that ion transport occurs into the lateral intracellular spaces but it remains unclear whether this leads to a hypertonic solution in these spaces causing an osmotically driven water absorption or if the process involves an osmotically linked isotonic secretion. These ion pumps are reversible, for water is absorbed during the interdigestive phase but fluid is secreted into the lumen during digestion or in the presence of disease. Appropriate neural stimulation can increase or decrease fluid absorption from the lumen while vasoactive intestinal peptide or secretin promote fluid secretion, probably mediated by prostaglandins leading to raised cyclic AMP acting at the cellular level. Immediate control may depend on intracellular Ca2+ which activates a calmodulin-protein kinase, phosphorylating the counter ion transporters to downregulate their activity. Failure of this regulatory process may explain the initial increase in bile concentrating potential seen in the development of gallstones although the mechanism of such failure remains unknown. More concentrated bile increases movement of biliary compounds into gallbladder epithelial cells which alter gallbladder function in a complex manner. Secondary bile acids are raised in gallstone disease and increase permeability of the gallbladder epithelium to molecules including cholesterol. This cholesterol absorbed from the lumen may have paramount importance to gallbladder function. Raised biliary cholesterol reduces gallbladder motility, possibly by increasing the amount of cholesterol in gallbladder muscle membranes and reducing contraction in response to cholecystokinin. However, increased secondary bile acids are also associated with an alteration in phospholipid acyl groups which may alter ion transport activity and/or cholesterol solubility within the micelle/vesicle. As the acyl groups show increased arachidonate levels the production of prostaglandins could be raised, although currently it is not known if this phospholipid arachidonate enters the epithelial cells. In addition, gallbladder inflammation is associated with raised phospholipase A2 activity, leading to formation of fatty acids and lysophospholipid which causes membrane damage. The fatty acids are likely to displace cholesterol from the micelle but may also act directly on the epithelium, possibly increasing prostaglandin production and thus stimulating mucin secretion. Increased mucin secretion is seen early in gallstone disease but the evidence presently available cannot determine if this is a causative factor. PMID- 9330350 TI - Characterization of glycoproteins in the epithelial cells of human and other mammalian gallbladder. A review. AB - The mammalian gallbladder mucosa is lined by a simple columnar epithelium. Typical surface epithelial cells (principal cells) contain short microvilli, secretory granules, dense bodies, mitochondria and Golgi apparatus. Dense bodies are thought to be lysosomes. Secretory granules contain mucous glycoproteins which are released to the lumen by exocytosis. Oligosaccharide side chains of mucous glycoproteins may provide a favorable environment for nucleation of cholesterol in gallstone formation; therefore they have been studied during the past decades. Histochemical techniques allow the in situ identification of carbohydrates at both the cellular and subcellular levels. The oligosaccharide chains of principal cell mucous glycoproteins have been studied by classical histochemical techniques (PAS, alcian blue, HID, etc). These techniques indicate that mammalian gallbladder mucous glycoproteins are heavily sulphated, whereas sialic acid residues are scarce. Neutral mucins have not been described in the mammalian gallbladder. Electron microscopic studies have located the oligosaccharide chains in secretory granules and Golgi apparatus. More recently, lectins (molecules which specifically recognize and bind with different saccharides or saccharide sequences) have been applied for the intracellular localization of carbohydrate residues. Lectin histochemistry has detected fucose, galactose, N-acetylglucosamine, N-acetylgalactosamine and N-acetylneuraminic acid residues in mucous granules, Golgi apparatus and apical membrane of human principal cells. Mannose residues were observed only in dense bodies. The combined use of deglycosylation procedures and lectin histochemistry has revealed a variety of terminal sequences in oligosaccharide chains of gallbladder mucous glycoproteins: Neu5Ac(alpha 2-3)Gal(beta 1-3)GalNAc, Neu5Ac(alpha 2-3)Gal(beta 1 4)GlcNAc and Gal(beta 1-4)GlcNAc. This technology also suggested the occurrence of N-linked oligosaccharides in the dense bodies of principal cells. Mucous granules mainly contained mucin-type O-linked oligosaccharides although some N linked chains have also been detected. Gallstone formation is probably a complex process depending on multiple factors. Mucous glycoproteins are one of the factors involved in this process. Histochemical methods offer an excellent research tool for the characterization of glycoproteins in the epithelial cells of the gallbladder, thus contributing to the elucidation of the pathophysiology of gallstone formation. PMID- 9330352 TI - Ultrastructural aspects of human gallbladder epithelial cells in cholelithiasis: production of anionic mucus. AB - The surface epithelium of 28 gallbladders removed during elective cholecystectomies and pathology collection was studied ultrastructurally. Focusing on 10 of the 28 cases that were diagnosed as cholecystitis, we found that the epithelium displayed numerous apical mucous granules and bulging apical apices. Mucous granule changes included 1) hyperproduction of secretory granules of neutral type containing an electron-dense proteinaceous spherule, similar to that described in other mucus-producing glands of the digestive system, and 2) production of anionic, osmiophilic secretory mucus. Other alterations of the surface epithelial cells included the production of bizarre surface appendages resembling primitive cilia without axoneme and epithelial excrescences. PMID- 9330353 TI - Organization of the blood and lymphatic microvasculature of the gallbladder in the guinea pig: a scanning electron microscopic study. AB - The organization of the blood and lymphatic microvessels of the gallbladder in the guinea pig is demonstrated by scanning electron microscopy (SEM) of vascular corrosion casts, and SEM of KOH-macerated tissues. In the lamina propria of the gallbladder, there is a dense network of subepithelial capillaries. The network is supplied by the arterioles that come off the arterial plexus located deep in the lamina propria. The network gathers into the postcapillary venules continuous with the collecting venular plexus located immediately below the subepithelial capillary network. The precapillary arterioles are sparsely surrounded by a single layer of circularly oriented extensions of smooth muscle cells. The terminal arterioles are endowed with circularly oriented fusiform smooth muscle cells. The nervous plexus is also noticed along the terminal arterioles. The capillaries are embraced by flat prolongations of pericytes. The postcapillary venules are sparsely surrounded by stellate pericytes and the collecting venules are sparsely surrounded by elongated or branched spindle-shaped, primitive smooth muscle cells which extend their long process in various directions along the vascular wall. The lymphatics are mostly located in the subserosal layer. The tips of the initial lymphatics are closed by endothelial cells, although there are frequently some gaps between them. The thin flaps of the lymphatic endothelial cells overlap or interdigitate with each other. The luminar surfaces of the lymphatics show oval nuclear protrusions, while the abluminal surfaces showed numerous microfolds except for the oval and flat nuclear portions. The lymphatics possess neither smooth muscle cells nor pericytes. PMID- 9330354 TI - Nociceptin-like immunoreactivity in the rat dorsal horn and inhibition of substantia gelatinosa neurons. AB - Nociceptin, also referred to as orphanin FQ, is believed to be the endogenous ligand for the ORL1. Nociceptin, when injected intracerebroventricularly to mice, produced hyperalgesia in behavioral tests. Recent studies have demonstrated the presence of ORL1 transcript in the spinal cord, and ORL1-like immunoreactivity has been localized to nerve fibers and somata throughout the spinal cord. Here, we report the localization of nociceptin-like immunoreactivity to fiber-like elements of the superficial layers of the rat dorsal horn by immunohistochemical techniques. Whole-cell recordings from substantia gelatinosa neurons in transverse lumbar spinal cord slices of 22-26-day-old rats showed that exogenous nociceptin at low concentrations (100-300 nM) depressed excitatory postsynaptic potentials evoked by stimulation of dorsal rootlets without causing an appreciable change of resting membrane potentials and glutamate-evoked depolarizations. At a concentration of 1 microM, nociceptin hyperpolarized substantia gelatinosa neurons and suppressed spike discharges. The hyperpolarizing and synaptic depressant action of nociceptin was not reversed by the known opioid receptor antagonist naloxone (1 microM). Our result provides evidence that nociceptin-like peptide is concentrated in nerve fibers of the rat dorsal horn and that it may serve as an inhibitory transmitter within the substantia gelatinosa. PMID- 9330355 TI - Brainstem-diencephalo-septohippocampal systems controlling the theta rhythm of the hippocampus. AB - We present a new model for the generation of theta rhythm of the hippocampus. We propose that theta at CA1 involves extracellular current fluxes produced by alternating depolarizing and hyperpolarizing membrane potential fluctuations of large populations of hippocampal pyramidal cells. Pyramidal cells are, in turn, controlled by rhythmically bursting cholinergic and GABAergic cells of the medial septum/vertical limb of the diagonal band. We postulate that septal cholinergic and GABAergic rhythmically bursting cells fire in relative synchrony; their coordinated burst discharge (burst mode) drives the positive-going phase of intracellular theta and associated firing of pyramidal cells; their synchronized pauses (interburst mode) give rise to the negative-going phase of intracellular theta and an inhibition of pyramidal cells. We further demonstrate that the theta rhythm is controlled by a network of cells extending from the brainstem to the septum/hippocampus. During theta, tonically discharging cells of the nucleus reticularis pontis oralis activate neurons of the supramammillary nucleus; the supramammillary nucleus, in turn, converts this steady barrage into a rhythmical pattern of discharge which is relayed to GABAergic/ cholinergic rhythmically bursting cells of the medial septum. The septal rhythmically bursting cells modulate subsets of hippocampal interneurons and principal cells in the generation of the theta rhythm. We review evidence showing that the serotonin containing neurons of the median raphe nucleus desynchronize the hippocampal electroencephalogram, presumably by disrupting the rhythmical discharge of septal cholinergic and GABAergic neurons. Finally, we summarize recent work indicating that the theta rhythm is critically involved in memory functions of the hippocampus and that its disruption (electroencephalographic desynchronization) may block or temporarily suspend mnemonic processes of the hippocampus. PMID- 9330356 TI - Metabotropic glutamate receptor activation modulates epileptiform activity in the hippocampus. AB - Synchronous neuronal activity that resembles interictal epileptiform discharges occurs in hippocampal slices if there is an imbalance of inhibitory and excitatory synaptic activity. Antagonists of the GABAA receptor and agonists of the ionotropic glutamate receptors are convulsants that produce epileptiform discharges in hippocampal slices. We evaluated the effects of activation of the metabotropic class of glutamate receptors on epileptiform activity produced by convulsants. The metabotropic glutamate agonist (+/-)-1-aminocyclopentane-trans 1,3-dicarboxylic acid (ACPD, 30-100 microM) accelerated the rate of interictal epileptiform discharges produced by either bicuculline methiodide or 4 aminopyridine and had minimal effects on discharges produced by high [K+]o. The increase in rate was associated with a significant decrease in the amplitude and duration of the afterhyperpolarization that follows the paroxysmal depolarizing shift, the intracellular correlate of the interictal epileptiform discharge. A modest increase in input resistance (approximately 10%) accompanied the rate increase. beta-adrenergic or muscarinic agonists, neurotransmitters that also decrease the afterhyperpolarization, acted synergistically with ACPD (100 microM) to increase the control rate of bicuculline-induced interictal discharges by more than eight-fold. Antagonists of beta-adrenergic or muscarinic receptors reduced, but did not block, the acceleration of bicuculline-induced discharge rate produced by 30 microM ACPD. The results show that metabotropic glutamate receptors enhance the rate of interictal epileptiform discharges produced by bicuculline or 4-aminopyridine. ACPD had no effect on interictal epileptiform activity induced by high [K+]o, a finding that may indicate that in high [K+]o conditions the metabotropic receptor is activated or that the effects of high [K+]o already reduced the effect of depolarizing currents that are enhanced by ACPD. The acceleration in interictal discharge rate was associated with a reduction in the afterhyperpolarization that follows the paroxysmal depolarizing shift and this action appears to be important in determining the synchronization of neurons and the rate of interictal epileptiform discharges. Furthermore, interaction between mGluR activation and either muscarinic or beta-adrenergic activation may be important for seizure generation. PMID- 9330357 TI - Electroresponsiveness of medial entorhinal cortex layer III neurons in vitro. AB - The entorhinal cortex funnels sensory information from the entire cortical mantle into the hippocampal formation via the perforant path. A major component of this pathway originates from the stellate cells in layer II and terminates on the dentate granule cells to activate the hippocampal trisynaptic circuit. In addition, there is also a significant, albeit less characterized, component of the perforant path that originates in entorhinal layer III pyramidal cells and terminates directly in area CA1. As a step in understanding the functional role of this monosynaptic component of the perforant path, we undertook the electrophysiological characterization of entorhinal layer III neurons in an in vitro rat brain slice preparation using intracellular recording techniques with sharp micropipettes and under current-clamp conditions. Cells were also intracellularly injected with biocytin to assess their pyramidal cell morphology. Layer III pyramidal cells did not display either the rhythmic subthreshold membrane potential oscillations nor spike-cluster discharge that characterizes the spiny stellate cells from layer II. In contrast, layer III pyramidal cells displayed a robust tendency towards spontaneous activity in the form of regular tonic discharge. Analysis of the voltage-current relations also demonstrated, in these neurons, a rather linear membrane voltage behaviour in the subthreshold range with the exception of pronounced inward rectification in the depolarizing direction. Depolarizing inward rectification was unaffected by Ca(2+)-conductance block with but was abolished by voltage-gated Na(+)-conductance block with tetrodotoxin, suggesting that a persistent Na(+)-conductance provides much of the inward current sustaining tonic discharge. In addition, in the presence of tetrodotoxin, an intermediate threshold (approximately -50 mV) Ca(2+)-dependent rebound potential was also observed which could constitute another pacemaker mechanism. A high-threshold Ca(2+)-conductance was also found to contribute to the action potential as judged by the decrease in spike duration towards the peak observed during Ca(2+)-conductance block. On the other hand, Ca(2+)-conductance block increase spike duration at the base and abolished the monophasic spike afterhyperpolarization. Analysis of the input-output relations revealed firing properties similar to those of regularly spiking neocortical cells. Current-pulse driven spike trains displayed moderate adaptation and were followed by a Ca(2+) dependent slow afterhyperpolarization. In summary, the intrinsic electroresponsiveness of entorhinal layer III pyramidal cells suggest that these neurons may perform a rather high-fidelity transfer function of incoming neocortical sensory information directly to the CA1 hippocampal subfield. The pronounced excitability of layer III cells, due to both Na+ and Ca2+ conductances, may also be related to their tendency towards degeneration in epilepsy. PMID- 9330358 TI - Sex-dependent effects of formalin and restraint on c-Fos expression in the septum and hippocampus of the rat. AB - In the present study we have demonstrated that the same aversive stimulus induces different patterns of expression of transcription factors in the hippocampus and septum of male and female rats. We have investigated by immunohistochemistry the effects of a persistent painful stimulus and restraint stress on c-Fos expression in the hippocampus and septum of male and female rats. Subjects were randomly assigned to one of three experimental groups: (i) untreated controls, (ii) subcutaneous injection with formalin (50 microliters, 10%) in the right hindpaw, or (iii) immobilization in an adjustable restrainer. Formalin-treated and restrained animals were killed 90 min after the beginning of treatment. In both male and female rats, unilateral injection of formalin induced bilateral c-Fos expression in the hippocampus, but the number of labeled neurons was two-fold higher in females than in males. Restraint stress was not effective in c-Fos induction in the hippocampus of both sexes. In the septum, both treatments increased c-Fos, but this increase tended to be greater in males than females. Previous experiments have consistently shown that male and female rats react differently to aversive stimulation. The present findings suggest that hormonal and behavioral differences between the sexes are accompanied by genetic modifications in those brain areas involved in cognition and emotion. PMID- 9330359 TI - Calcium-mediated intracellular messengers modulate the serotonergic effects on axonal excitability. AB - We carried out experiments to investigate the mechanisms of serotonin-induced axonal excitability changes using isolated dorsal columns from young (seven to 11 day-old) Long-Evan's hooded rats. Conducting action potentials were activated by submaximal (50%) and supramaximal constant current electrical stimuli and recorded with glass micropipette electrodes. In experiment 1, to study Ca(2+) mediated mechanisms, we superfused the preparations with Ringer solutions containing varying Ca2+ concentrations. Following superfusion with Ca(2+)-free Ringer solution for 4 h, we tested initial responses to serotonin agonists. Studies then were repeated after preparations had been washed for 1 h with Ringer solution containing 1.5 mM Ca2+ and 1.5 mM Mg2+. After 4 h superfusion of Ca(2+) free Ringer solution, quipazine (a serotonin2A agonist, 100 microM) did not induce significant axonal excitability changes (amplitude change of 1.4 +/- 1.3%, percentage of predrug control level, +/-S.D., n = 6). A 100 microM concentration of 8-hydroxy-dipropylaminotetralin (a serotonin1A agonist) reduced response amplitudes by 36.3 +/- 4.2% (+/-S.D., P < 0.0005, n = 7) and prolonged latencies by 22.3 +/- 4.3% (+/-S.D., P < 0.0005, n = 7). Application of serotonin (100 microM) decreased amplitudes by 6.6 +/- 5.0% (+/-S.D., P < 0.05, n = 6). Extracellular calcium concentration ([Ca2+]e) was measured at various depths in the dorsal column with ion-selective microelectrodes. Four hours' superfusion with Ca(2+)-free Ringer solution reduced [Ca2+]e to less than 0.1 mM in dorsal columns. In 1.5 mM Ca2+ Ringer solution, quipazine increased the amplitudes by 38.3 +/- 5.8% (P < 0.0005, n = 6). Likewise, serotonin increased the amplitudes by 13.8 +/- 4.9% (P < 0.005, n = 6). In contrast however, 8-hydroxy dipropylaminotetralin still reduced amplitudes by 35.0 +/- 6.4% (P < 0.0005, n = 7) and prolonged latencies by 24.1 +/- 4.5% (P < 0.0005, n = 7). In experiment 2, we investigated calcium-dependent and cAMP-mediated protein kinase signalling pathways to evaluate their role as intracellular messengers for serotonin2A receptor activation. Two protein kinase inhibitors, 50 microM H7 (an inhibitor of protein kinase C and c-AMP dependent protein kinase) and 100 microM D-sphingosine (an inhibitor of protein kinase A and C) effectively eliminated the excitatory effects of the serotonin2A agonist. 100 microM cadmium (a Ca2+ channel blocker) also blocked the effects of quipazine. Neither these protein kinase inhibitors nor cadmium alone affected action potential amplitudes. These results suggest that replacing Ca2+ with Mg2+ blocks the excitatory effects of quipazine but does not prevent the inhibitory effects of 8-hydroxy-dipropylaminotetralin, and calcium-mediated protein kinase mechanisms modulate axonal excitability changes induced by serotonin and its agonist. PMID- 9330360 TI - Microinjections of muscimol into lateral superior colliculus disrupt orienting and oral movements in the formalin model of pain. AB - An important reaction in rodent models of persistent pain is for the animal to turn and bite/lick the source of discomfort (autotomy). Comparatively little is known about the supraspinal pathways which mediate this reaction. Since autotomy requires co-ordinated control of the head and mouth, it is possible that basal ganglia output via the superior colliculus may be involved; previously this projection has been implicated in the control of orienting and oral behaviour. The purpose of the present study was therefore, to test whether the striato-nigro tectal projection plays a significant role in oral responses elicited by subcutaneous injections of formalin. Behavioural output from this system is normally associated with the release of collicular projection neurons from tonic inhibitory input from substantia nigra pars reticulata. Therefore, in the present study normal disinhibitory signals from the basal ganglia were blocked by injecting the GABA agonist muscimol into different regions of the rat superior colliculus. c-Fos immunohistochemistry was used routinely to provide regional estimates of the suppressive effects of muscimol on neuronal activity. Biting and licking directed to the site of a subcutaneous injection of formalin (50 microliters of 4%) into the hind-paw were suppressed in a dose-related manner by bilateral microinjections of muscimol into the lateral superior colliculus (10-50 ng; 0.5 microliter/side); injections into the medial superior colliculus had little effect. Bilateral injections of muscimol 20 ng into lateral colliculus caused formalin-treated animals to re-direct their attention and activity from lower to upper regions of space. Muscimol injected unilaterally into lateral superior colliculus elicited ipsilateral turning irrespective of which hind-paw was injected with formalin. Oral behaviour was blocked when the muscimol and formalin injections were contralaterally opposed; this was also true for formalin injections into the front foot. Interestingly, when formalin was injected into the perioral region, injections of muscimol into the lateral superior colliculus had no effect on the ability of animals to make appropriate contralaterally directed head and body movements to facilitate localization of the injected area with either front- or hind-paw. These findings suggest that basal ganglia output via the lateral superior colliculus is critical for responses to noxious stimuli which entail the mouth moving to and acting on the foot, but not when the foot is the active agent applied to the mouth. The data also suggest that pain produces a spatially non-specific facilitation of units throughout collicular maps, which can be converted into a spatially inappropriate signal by locally suppressing parts of the map with the muscimol. PMID- 9330361 TI - Ischemic tolerance in hippocampal CA1 neurons studied using contralateral controls. AB - We induced ischemic tolerance unilaterally in gerbil hippocampus using the contralateral hippocampus as control. Ischemia for 2 min of right common carotid occlusion was reversible but sufficient to cause heat-shock protein 70 production in CA1 neurons. This pretreatment given four days prior to occlusion of both common carotids for 5 min, but not at longer preceding intervals, induced tolerance in right CA1 neurons. Neuroprotection was still evident two months after the 5 min occlusion. Adenosine triphosphate content and immunoreactive microtubule associate protein 2 in the hippocampus showed that the 5 min ischemic insult was essentially equal in both hemispheres. Repetitive pretreatments at two day intervals caused almost complete protection of CA1 neurons against subsequent 5 min ischemia, while a single pretreatment showed 80% protection. However, the increase in heat-shock protein 70 with repeated pretreatments was not significantly more than with one pretreatment. We concluded that true ischemic tolerance was induced by ischemic stress itself, was long-lasting, was not due to mitigation of subsequent ischemia, and was augmented by repetition without further increase of heat-shock protein 70. PMID- 9330362 TI - Sodium channel blockade unmasks two temporally distinct mechanisms of striatal dopamine release during hypoxia/hypoglycaemia in vitro. AB - Massive striatal dopamine release during cerebral ischaemia has been implicated in the resulting neuronal damage. Sodium influx is an early event in the biochemical cascade during ischaemia and blockade of sodium channels may increase resistance to ischaemia by reducing energy demand involved in compensation for sodium and potassium fluxes. In this study, we have determined the effects of opening and blockade of voltage-gated sodium channels on hypoxia/hypoglycaemia induced dopamine release. Slices of rat caudate nucleus were maintained in a slice chamber superfused by an oxygenated artificial cerebrospinal fluid containing 4 mM glucose. Ischaemia (hypoxia/hypoglycaemia) was mimicked by a switch to a deoxygenated artificial cerebrospinal fluid containing 2 mM glucose and dopamine release was measured using fast cyclic voltammetry. In drug-free (control) slices, there was a 2-3 min delay after the onset of hypoxia/hypoglycaemia followed by a rapid dopamine release event which was associated with anoxic depolarization. In slices treated with the Na+ channel opener, veratridine (1 microM), the time to onset of dopamine release was shortened (101 +/- 20 s, compared with 171 +/- 8 s in controls, P < 0.05). Conversely, phenytoin (100 microM), lignocaine (200 microM) and the highly selective sodium channel blocker, tetrodotoxin (1 microM) markedly delayed and slowed dopamine release vs paired controls. In the majority of cases, dopamine release was biphasic after sodium channel blockade: a slow phase preceded a more rapid dopamine release event. The latter was associated with anoxic depolarization. Neither the fast nor the slow release events were affected by pretreatment with the selective dopamine uptake blocker GBR 12935 (0.2 microM), suggesting that uptake carrier reversal did not contribute to these events. In conclusion, sodium channel antagonism delays and slows hypoxia/hypoglycaemia induced dopamine release in vitro. Furthermore, sodium channel blockade delays anoxic depolarization and its associated neurotransmitter release, revealing an earlier dopamine release event that does not result from reversal of the uptake carrier. PMID- 9330363 TI - Nicotinic antagonist administration into the ventral hippocampus and spatial working memory in rats. AB - Nicotinic acetylcholine receptors are important for maintaining optimal memory performance. In order to more fully characterize the involvement of nicotinic systems in memory, the contributions of nicotinic acetylcholine receptor subtypes were investigated. This study targeted the alpha 7 and alpha 4 beta 2 nicotinic receptors in the ventral hippocampus, an area known to be important for spatial working memory. Antagonists of alpha 7 and alpha 4 beta 2 receptors were locally infused into the ventral hippocampus of rats and the effects on memory were examined with the radial-arm maze. The subtype-specific competitive antagonists infused into separate groups of rats were methyllycaconitine citrate (an alpha 7 antagonist) and dihydro-beta-erythroidine hydrobromide (an alpha 4 beta 2 antagonist). Their effects on radial-arm maze performance were contrasted with the non-specific competitive antagonist, D-tubocurarine chloride. Significant deficits in radial-arm maze choice accuracy performance were found at 78.7 micrograms/side for methyllycaconitine and at 106.9 micrograms/side for dihydro beta-erythroidine. Increased response latency was also seen at these doses. Tubocurarine induced seizures at doses previously reported to have no effect. Wet dog shakes were seen in most rats at 0.1 microgram/side with tubocurarine, 26.3 micrograms/side with methyllycaconitine and 106.9 micrograms/side with dihydro beta-erythroidine. This study suggests that both alpha 7 and alpha 4 beta 2 nicotinic acetylcholine receptor subtypes are involved in working memory formation and that the hippocampus is a critical site for nicotinic cholinergic involvement in memory function, though the high doses of antagonists needed to produce the memory impairment may have had less than completely specific effects. PMID- 9330364 TI - Transforming growth factor-alpha's effects on astroglial-cholinergic cell interactions in the medial septal area in vitro are mediated by alpha 2 macroglobulin. AB - We reported previously that two epidermal growth factor receptor ligands, epidermal growth factor and transforming growth factor-alpha, inhibit medial septal cholinergic cell phenotypic expression (choline acetyltransferase and acetylcholinesterase activities) in vitro indirectly via (a) soluble molecule(s) released from astrocytes [Kenigsberg R. L. et al. (1992) Neuroscience 50, 85-97; Kenigsberg R. L. and Mazzoni I. E. (1995) J. Neurosci. Res. 41, 734-744; Mazzoni I. E. and Kenigsberg R. L. (1996) Brain Res. 707, 88-99]. In the present study, we found that this response to transforming growth factor-alpha is mediated, for the most part, by alpha 2-macroglobulin, a potent protease inhibitor with a wide spectrum of biological activities. In this regard, the effects of transforming growth factor-alpha on cholinergic cells can be blocked with immunoneutralizing antibodies raised against alpha 2-macroglobulin. Furthermore, western blot analysis reveals that although alpha 2-macroglobulin is present in conditioned media from control septal cultures, it is more abundant in those treated with transforming growth factor-alpha. In addition, exogenous alpha 2-macroglobulin, both in its native and trypsin-activated forms, can mimic transforming growth factor-alpha's effects on septal cholinergic cell expression. However, while the native antiprotease can slightly but significantly decrease choline acetyltransferase activity, trypsin-activated alpha 2-macroglobulin, in the nanomolar range, induces as marked a decrease in this enzyme activity as that noted with transforming growth factor-alpha. Furthermore, trypsin-activated alpha 2-macroglobulin, like epidermal growth factor/transforming growth factor-alpha, decreases choline acetyltransferase activity by arresting its spontaneous increase that occurs with time in culture, does so in a reversible manner and is not neurotoxic. In addition, trypsin-activated alpha 2-macroglobulin, in the nanomolar range, can affect choline acetyltransferase in a dual manner, up regulating it at low concentrations while down-regulating it at higher ones. These responses are identical in mixed neuronal-glial and pure neuronal septal cultures. Furthermore, when concentrations of trypsin-activated alpha 2 macroglobulin, which alone decrease choline acetyltransferase, are added simultaneously with nerve growth factor, they serve to potentiate the nerve growth factor-induced increase in enzymatic activity. As GABAergic cell expression is not affected by alpha 2-macroglobulin, it appears that the effects of this protease inhibitor on medial septal neuronal expression are neurotransmitter-specific. Finally, trypsin-activated but not native alpha 2 macroglobulin promotes a dose-dependent aggregation of the septal neurons. This change in morphology, however, is not related to those noted in choline acetyltransferase activity. In summary, these data suggest that the expression of alpha 2-macroglobulin in astroglia from the medial septal nucleus can be controlled by epidermal growth factor receptor ligands to impact the functioning of basal forebrain cholinergic neurons. PMID- 9330365 TI - Cellular and synaptic localization of the neuronal glutamate transporters excitatory amino acid transporter 3 and 4. AB - Glutamate transport is a primary mechanism for the synaptic inactivation of glutamate. Excitatory amino acid transporter 4 (EAAT4) is a novel glutamate transporter with properties of a ligand-gated chloride channel that was recently cloned from human brain. The present study was an investigation of the protein expression and cellular localization of EAAT4 in human and rat brain, and comparison with another neuronal glutamate transporter, EAAT3 (rabbit excitatory amino acid carrier 1; EAAC1). Regional immunoblot analysis of EAAT4, using a monospecific oligopeptide (carboxy-terminal) affinity-purified polyclonal antibody, revealed that the protein was restricted to the central nervous system. The EAAT4 protein was largely expressed in cerebellum, with a much lower expression in hippocampus, neocortex, striatum, brain stem and thalamus. Immunohistochemical studies showed intense EAAT4 immunoreactivity in the human and rat cerebellar Purkinje cells with a somatodendritic localization. Other brain regions including neocortex, hippocampus, striatum showed faint neuropil staining of EAAT4. Immunogold localization identified EAAT4 protein at plasma membranes of Purkinje cell dendrites and spines. In the hippocampus and neocortex, EAAT4 immunoreactivity was found mainly at small calibre dendrites. Rarely, EAAT4 immunoreactivity was found in astrocytic cell processes of forebrain. In the cerebellum, EAAT4 localization partly overlapped with the neuronal localization of EAAT3 (EAAC1). Immunoreactivity for EAAT3 was enriched in the somatodendritic compartment of the Purkinje cells like EAAT4, but EAAT3 was also found in Purkinje cell axons and in boutons in deep cerebellar nuclei, as well as in granular cells and stellate cells. Our results indicate that EAAT4 protein is largely localized to cerebellar cortex and lower levels of EAAT4 protein are present in forebrain by immunoblot and immunohistochemistry. Both neuronal glutamate transporter EAAT3 (EAAC1) and EAAT4 are located at somatodendritic compartment of Purkinje cells, and probably contribute to glutamate re-uptake mechanisms at Purkinje cell synapses. PMID- 9330366 TI - GABAA-receptor alpha-subunit is an essential prerequisite for receptor formation in vivo. AB - The mechanisms governing the assembly of alpha-, beta- and gamma-subunits to form GABAA-receptors are poorly understood. Here, we report that the alpha-subunit is essential for receptor assembly. In mice homozygous for a deletion on chromosome 7 spanning the alpha 5- and gamma 3-subunit genes, zolpidem-insensitive benzodiazepine binding sites, corresponding to GABAA-receptors containing the alpha 5-subunit, were absent in the hippocampus. This loss of alpha 5-GABAA receptor binding was also apparent as a 21% decrease in the total number of benzodiazepine binding sites in the hippocampus. In addition, immunoreactivity for the beta 2,3- and gamma 2-subunit was decreased exclusively in neurons which normally express the alpha 5-subunit, such as olfactory bulb granule cells and hippocampal pyramidal cells. In other brain regions of the mutants, the beta 2,3- and gamma 2-subunit staining was unaffected. Controls included two lines of mice homozygous for a shorter chromosomal deletion, that either included or excluded the gamma 3-subunit gene. These two lines were indistinguishable with regard to numbers of benzodiazepine binding sites and levels alpha 5-, beta 2,3- and gamma 2-subunit immunoreactivity, indicating that the lack of gamma 3-subunit gene did not contribute to the observed deficit in receptor formation. These results demonstrate that the absence of the alpha 5-subunit gene prevents the formation of the entire respective receptor complex in adult mouse brain. Thus, the alpha subunit, unlike the gamma 2-subunit, might play a major role in the assembly or targeting of GABAA-receptor complexes. PMID- 9330367 TI - Chronic cold stress alters the basal and evoked electrophysiological activity of rat locus coeruleus neurons. AB - In vivo extracellular single-unit recording techniques revealed that chronic cold stress significantly alters both the basal and the evoked electrophysiological activity of noradrenergic neurons in the locus coeruleus of the anaesthetized rat. Following 17-21 days of chronic cold exposure (5 degrees C), the single-unit activity of histologically-identified locus coeruleus neurons in chloral hydrate anaesthetized rats was recorded and analysed in terms of their basal firing rate and pattern of spike activity, as well as their response to footshock stimulation. There was no significant difference in the incidence of spontaneously active cells/electrode track between cold-stressed rats and control rats. However, the basal spike activity of locus coeruleus cells recorded from cold-stressed rats differed significantly from that of control rats along two dimensions: i) they displayed significantly higher basal firing rates (mean = 1.88 Hz vs 1.20 Hz, respectively); and ii) they frequently exhibited spontaneous burst-firing activity that was not observed in control rats (observed in 15/17 cold-stressed rats vs 1/26 control rats). The evoked spike activity of locus coeruleus cells in cold-stressed rats also differed significantly from that of control rats along two dimensions: i) they were more likely to respond to footshock stimulation (mean = 90.3% vs 74.4%, respectively); and ii) these responses were more likely to consist of multispike bursts of action potentials (mean = 8 bursts/50 stimulations vs 1 burst/50 stimulations, respectively). These results indicate that alterations in the electrophysiological activity of noradrenergic locus coeruleus neurons may contribute to the phenomenon of stress induced sensitization of norepinephrine release that is thought to underlie some of the neuropathological changes that accompany long-term stress. PMID- 9330368 TI - Distribution of activated neurons in the rabbit brain following a volume load. AB - Immunohistochemical detection of the protein, Fos, was used to identify neurons in the brain activated following a volume load. The plasma expanders, Haemaccel and 6% dextran, were infused intravenously in conscious rabbits for 60 min. Compared to control animals both stimuli significantly increased right atrial pressure but had no effect on blood pressure. Heart rate was significantly elevated with dextran only. Volume expansion with Haemaccel also reduced renal sympathetic nerve activity by about 50% from the pre-infusion resting level. Ninety minutes after the start of the infusion, the rabbits were perfusion fixed and the distribution of Fos-positive cell nuclei was examined. Following Haemaccel infusion there were significant increases in the number of Fos-positive cell nuclei in the organum vasculosum of the lamina terminalis, parvocellular paraventricular nucleus and in specific rostrocaudal levels of the nucleus tractus solitarius and ventrolateral medulla. Following dextran similar effects were observed in the medulla but Fos-positive cell nuclei were not significantly elevated above controls in the forebrain. After Haemaccel or dextran areas such as the supraoptic nucleus, the magnocellular paraventricular nucleus, the bed nucleus of the stria terminalis, diagonal band of Broca and amygdala either did not produce Fos or were not consistently different from the control group. The results suggest that specific brain regions, that are known to be important in cardiovascular control, are activated by a volume load. These areas are likely to play an important role in the reflex responses initiated by that particular stimulus. PMID- 9330370 TI - Unilateral fetal mesencephalic grafts in intact rats reduce amphetamine-induced dopamine release in both striata. An in vivo voltammetric study. AB - This study investigated amphetamine-induced striatal dopamine release after intraventricular unilateral fetal mesencephalic grafts in otherwise intact rats. Dopamine was monitored in vivo by differential pulse voltammetry. In grafted animals, amphetamine-induced dopamine release was decreased compared to sham grafted, age-matched controls. This decrease was observed in the grafted as well as in the contralateral striatum five months after intraventricular grafting. There was no measurable effect of the graft on the amphetamine-induced rotational behaviour. Our results exceed former observations reporting decreased amphetamine induced dopamine release in the contralateral striatum of 6-hydroxydopamine lesioned and unilaterally-grafted rats which had been attributed to a reduction of dopamine transporters. Furthermore, it was shown that concerning this effect ventral mesencephalic grafts are independent of a previous 6-hydroxydopamine lesion. PMID- 9330369 TI - Role of metabotropic glutamate receptors in the depression of GABA-mediated depolarization of frog primary afferent terminals. AB - Sucrose gap recordings from the dorsal roots of isolated, hemisected frog spinal cords were used to determine the effects of metabotropic L-glutamate receptor activation on primary afferent terminals by (+/-)-1-amino-trans-1,3-cyclopentane dicarboxylic acid (t-ACPD). Dorsal root potentials evoked by ventral root volleys were significantly reduced by t-ACPD (30 microM), as were GABA- and muscimol induced afferent terminal depolarizations. The effects of t-ACPD on GABA depolarizations depended upon activation of group I metabotropic glutamate receptors, i.e. the effects were blocked by the group I/II antagonist (RS)-alpha methyl-4-carboxyphenylglycine, but not by the group II antagonist alpha-methyl (2S,3S,4S)-alpha-(carboxycyclopropyl)-glycine or the group III antagonist alpha methyl-(S)-2-amino-4-phosphonobutyrate and were mimicked by the group I agonist 3,5-dihydroxyphenylglycine but were not mimicked by the group III agonist (S)-2 amino-4-phosphonobutyrate. Increasing the intracellular concentration of 3'-5' cyclic adenosine monophosphate with 8-bromo-cAMP, forskolin, and 3-isobutyl-1 methylxanthine significantly reduced GABA depolarizations, but the protein kinase inhibitors Rp-adenosine 3,5-cyclic monophosphothioate triethylamine and N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide did not alter t-ACPD's depression of GABA depolarizations. The actions of t-ACPD on GABA depolarizations were neither mimicked nor blocked by phorbol-12-myristate 13-acetate, thapsigargin, staurosporine, or arachidonic acid, presumptive indications that the effects of t-ACPD did not involve phosphoinositide hydrolysis, the release of Ca2+ from intracellular stores, or the formation of arachidonate. t-ACPD's effects on GABA depolarizations were blocked by 20 mM Mg2+, the broad spectrum L glutamate antagonist kynurenate, and the selective N-methyl-D-aspartate antagonist D(-)-2-amino-5-phosphonovaleric acid, but not by the non-N-methyl-D aspartate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione. Low concentrations of N-methyl-D-aspartate (10 microM) mimicked the effect of t-ACPD on GABA responses. These results suggest that t-ACPD's depression of GABA depolarizations involves an indirect, three-stage mechanism that includes activation of Group I metabotropic glutamate receptors on interneurons and/or on afferent terminals, the release of L-glutamate from the latter structures, and the activation of N methyl-D-aspartate receptors on primary afferent terminals. The depression of GABA depolarizations caused by the release of L-glutamate from afferent terminal and/or interneurons leads to a block of presynaptic inhibition (produced in the frog spinal cord by GABA) resulting in a positive feed-forward amplification of reflex transmission. PMID- 9330371 TI - Glial cell line-derived neurotrophic factor attenuates the excitotoxin-induced behavioral and neurochemical deficits in a rodent model of Huntington's disease. AB - The present study determined the effects of intraventricularly administered glial cell line-derived neurotrophic factor on the behavioral and neurochemical sequelae of unilateral excitotoxic lesions of the striatum. Distinct asymmetrical rotational behavior in response to peripheral administration of amphetamine (5 mg/kg) was noted one and two weeks following injections of quinolinic acid (200 nmol) into two sites in the left striatum. In rats given a single intraventricular injection of glial cell line-derived neurotrophic factor (10 1000 micrograms) 30 min before the toxin, amphetamine-induced rotational behavior was significantly attenuated. Analysis of Nissl-stained coronal sections showed marked neuronal loss in the striatum ipsilateral to the quinolinic acid injections, which was at least partially prevented by glial cell line-derived neurotrophic factor D1 and D2 dopamine binding sites in the striatum, the majority of which are localized to subpopulations of GABAergic neurons, were decreased to a similar extent by quinolinic acid. Moreover, the reduction was attenuated by glial cell line-derived neurotrophic factor treatment to a similar degree, suggesting that the two subpopulations of GABAergic striatal output neurons are equally vulnerable to excitotoxic damage. Concomitant changes in neurotransmitter function as a result of the lesion were also observed: [3H]GABA uptake into striatal target tissues (globus pallidus and substantia nigra) was considerably reduced in the lesioned compared to the contralateral unlesioned tissues, as were [3H]choline and [3H]dopamine uptake into striatal synaptosomes. Similarly, striatal choline acetyltransferase activity was decreased by the lesion. Decrements in neuropeptide levels of similar magnitude were evident ipsilateral to the lesion; substance P, met-enkephalin and dynorphin A contents in the globus pallidus and substantia nigra were significantly reduced. Striatal somatostatin and neuropeptide Y levels were not altered. All of the neurochemical deficits induced by striatal quinolinic acid lesions were attenuated by intraventricular delivery of glial cell line-derived neurotrophic factor. Continuous intraventricular infusion of this trophic factor (10 micrograms/day) over a two-week period did not afford notable improvement compared to the single injection of 10 micrograms. In contrast, continuous infusion of brain-derived neurotrophic factor (10 micrograms/day) directly into the striatum did not affect any of the neurochemical parameters studied. However, neurotrophin-3 (10 micrograms/day) delivery into the striatum significantly increased [3H]GABA uptake, but only modestly affected [3H]choline uptake. The results indicate that glial cell line-derived neurotrophic factor counteracts neuronal damage induced by a striatal excitotoxic insult and support its potential use as a treatment for central nervous system disorders that may be a consequence of excitotoxic processes, such as Huntington's disease. PMID- 9330372 TI - The role of long-term potentiation in persistent epileptiform burst-induced hyperexcitability following GABAA receptor blockade. AB - Persistent hyperexcitability follows synchronized bursting induced in the CA3 region of hippocampal slices by perfusion with high concentrations (2000 IU/ml) of the GABAA antagonist, penicillin. This hyperexcitable state is characterized by: i) slow recovery from bursting following penicillin washout; ii) persistent "post-burst" field potential oscillations and iii) increased probability of spontaneous bursting with ordinarily sub-convulsant doses of GABAA antagonists. An N-methyl-D-aspartate-independent type of long-term potentiation of alpha-amino 3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate excitatory postsynaptic potentials occurred following bursting. However, similar increases in excitatory postsynaptic potential magnitude also occurred after a subconvulsant dose of penicillin (500 IU/ml) which did not produce the other features of persistent hyperexcitability. Furthermore, long-term potentiation either increased or remained stable after bursting stopped, whereas, post-burst oscillations gradually diminished with time. Low doses of the AMPA/kainate antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, which restored the potentiated excitatory postsynaptic potentials to control levels, reduced but did not eliminate the post-burst oscillation. Tetanus-induced long-term potentiation did not reproduce the hyperexcitable state seen after bursting. These findings indicate that the epileptiform bursting caused by blocking GABAA-mediated inhibition induces long-term potentiation which is partially responsible for persistent burst-induced hyperexcitability but is not sufficient to entirely explain it. The hippocampus which is critical for normal memory is also frequently the generator of intractable epileptic seizures. Seizure-like discharges in the hippocampus induced long-lasting increases in synaptic efficacy similar to those thought to underlie normal memory. This form of long-term potentiation contributed to the network oscillations characteristics of the hyperexcitable state persisting after epileptiform activity but was not sufficient to entirely explain them. Epileptic seizures may engage normal memory mechanisms which increase neuronal excitability and predispose the hippocampal network to further seizures. This may, in part, account for the propensity for hippocampal seizure foci to become intractable. PMID- 9330373 TI - Developmental changes in calpain activity, GluR1 receptors and in the effect of kainic acid treatment in rat brain. AB - The cellular distribution of calpain activation and glutamate receptor 1 (GluR1) subunits of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and their alterations following kainic acid-induced seizure were evaluated during postnatal development using antibodies specific for spectrin breakdown product and the C-terminus of GluR1 subunits. In the first postnatal week, most brain regions exhibited high levels of calpain activity that progressively decreased during the following weeks. The highest levels of spectrin breakdown product immunoreactivity were observed in the somata and proximal dendrites of hippocampal pyramidal cells, non-pyramidal neurons in stratum oriens, and cortical neurons. In general, during the first two postnatal weeks, kainic acid treatment induced a decrease in spectrin breakdown product immunoreactivity in neuronal cell bodies and an increase in dendritic fields. Obvious elevation in spectrin breakdown product immunoreactivity in selective non-pyramidal cells in stratum oriens started at postnatal day 14, and was further evidenced by postnatal day 21. Likewise, massive calpain activation in subpopulations of neurons in some thalamic nuclei, amygdala, and pyriform cortex was observed after the third postnatal week. GluR1 subunits were highly expressed throughout the forebrain in the first postnatal week, further increased during the second postnatal week, decreased thereafter, and reached adult levels after postnatal day 21. In cortex, intense GluR1 immunostaining was found in the somata and proximal processes of pyramidal and non-pyramidal neurons, with the non-pyramidal neurons in layers IV through VI exhibiting the densest immunolabelling. In the first two postnatal weeks, the somata of hippocampal pyramidal neurons exhibited intense GluR1 immunostaining that became more dendritic in the subsequent developmental period. While hilar cells exhibited a similar developmental pattern as CA regions, the molecular layer of dentate gyrus exhibited weak immunoreactivity from postnatal day 7 to postnatal day 14. The early increase in GluR1 immunoreactivity in hippocampal pyramidal layer following kainic acid treatment occurred throughout the developmental period, while the later decrease in CA regions, amygdala, and pyriform cortex was observed only in postnatal day 21 animals. The combined immunocytochemical studies of spectrin breakdown product localization and GluR1 expression indicate that calpain activation might play an important role in synaptic formation, developmental regulation of synaptic plasticity, and neuronal vulnerability to excitotoxicity during postnatal development. Moreover, calpain-mediated modulation of alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid receptors might underlie these processes. PMID- 9330374 TI - Expression of calretinin in diverse neuronal populations during development of rat hippocampus. AB - The prenatal and postnatal expression of calretinin was studied in hippocampus of the rat using immunohistochemical procedures. Calretinin was detected as early as embryonic day 15 in the primordial hippocampus where calretinin-containing neurons and fibres were localized to the primitive plexiform layer. Upon emergence of the hippocampal plate (the prospective stratum pyramidale), large numbers of immunopositive multipolar cells were observed in the marginal zone. Fewer cells with fusiform cell bodies were observed bordering the hippocampal plate and subplate. During the perinatal period (embryonic day 20 to postnatal day 0), large numbers of immunoreactive pyramidal-like neurons were observed at the margin of the hippocampal plate with the subplate. At this same time, many calretinin-containing neurons with irregularly shaped dendrites were observed in stratum radiatum. Soon after birth (postnatal day 3), the calretinin immunoreactivity of both these later cell types rapidly declined and a new population of calretinin-immunopositive cells emerged, the Cajal-Retzius cells of stratum lacunosum-moleculare and the dentate gyrus. The Cajal-Retzius cells rapidly matured but disappeared by the second postnatal week. During the second postnatal week, calretinin interneurons of the adult hippocampal formation began to appear. Their immunoreactivity increased by postnatal day 15, when the number of calretinin-immunopositive interneurons in area CA1 and stratum radiatum of CA3 exceeded that of the adult. At this time, the soma and proximal dendrites of many calretinin interneurons were found to contact each other. The frequency of such cellular appositions decreased in adulthood. The results presented here show that calretinin immunohistochemistry can be very useful in recording the development of subpopulations of hippocampal neurons that are present during distinct embryonic and postnatal periods. Although some neuronal types may exist only briefly during hippocampal development, others appear to express calretinin transiently during restricted phases of neuronal differentiation. Surprisingly, this includes some hippocampal pyramidal cells. However, even as the adult pattern of immunostaining emerges in week 2, morphological refinement of interneurons continues to take place, which eventually leads to the population of calretinin-containing interneurons of the mature hippocampus. PMID- 9330375 TI - Spatial distribution of field potential profiles in the cat cerebellar cortex evoked by peripheral and central inputs. AB - The present study was designed to characterize the spread of excitation within the frontal plane of the cat cerebellar cortex following different types of stimuli. In particular, experiments were performed to determine whether the spread of excitation evoked by mossy fibre inputs proceeds primarily along the parallel fibres ("beam-like" spread) or whether these inputs activate non propagated foci ("patches") in the cerebellar cortex. Field potentials were recorded within a frontal plane as a medial to lateral array at different depths in parallel tracks. The recordings were made following electrical stimulation of different forelimb nerves and functionally related areas of the sensorimotor cortex as well as during passive paw movements. The resulting spatial grid of responses provides discrete spatio-temporal information reflecting the activation of specific cerebellar afferents and the neuronal interactions they evoke. The method employed demonstrates the spatial distribution of the temporal sequence of excitability changes throughout all the cerebellar cortical layers. In general, the characteristics of the responses in the intermediate cerebellar cortex depended on the source of the signals. Activity patterns evoked by peripheral nerve stimulation showed more clustered foci compared with those following electrical stimulation of functionally related areas of the sensorimotor cortex. The centrally evoked profiles were generally more homogeneous. The largest number of foci were observed following passive movements around the wrist joint. The spread of excitation in the vertical direction was evaluated by the spatial shift of the line of reversal of the N3/P2-potential (zero-isopotential line). Lines of reversal for peripherally-evoked activity patterns were approximately 90 microns closer to the molecular layer than those evoked by central stimulation in animals in which recordings have been performed in lobule Vc. The opposite was found for recordings in lobule Vb, where potential reversals following peripheral stimulation were located 40 microns deeper than those evoked following central stimulation. Cortical inputs resulted in a more proximal activation of lobule Vc Purkinje cell dendrites than in lobule Vb. This type of input processing thus seems to be lobule dependent. A beam-like spread of excitation could not be demonstrated. For both climbing fibre and mossy fibre afferent systems multiple foci were found in the frontal plane. The foci due to mossy fibre activation arose from the granular layer and expanded vertically to the molecular layer. For the climbing fibre system the foci were restricted to the molecular layer, where they merged to form a superficial band of activation. Although the data presented in this paper favour a focal distribution of activity, they do not exclude beam like propagation along the parallel fibres, because of the difficulty of detecting this pattern in response to the stimuli. The "beam"- and "patch"-like hypotheses need not be mutually exclusive. Each could contribute to a specific stage of the temporal-spatial processing in the cerebellar cortex in a functional and task-specific manner. PMID- 9330377 TI - [Preoperative estimation of liver injury and operative risk]. AB - We report on preoperative estimation of operative risk, principally rating liver injury in chronic liver disease, c.g., chronic hepatitis and cirrhosis, for liver resection and general surgery. Regarding general surgery, elective and standard operation are possible in Child A and operations with measures to lessen intraoperative blood loss and lymphadenectomy in Child B, but in Child C, surgery is limited to emergency palliative operations, and conservative treatment methods must chosen. In liver resection and major surgery it is important to estimate extent of liver resection and operative risk, primarily by R15 and KICG, and make an overall judgment based on fibronectin, hyaluronic acid, sinusoidal endothelial cell function measured by thrombomodulin, sigma IRI in the 75g OGTT, Fischer's ratio and other indices of lipid metabolism. Generally, surgery limits are: KICG, 40%. Conventional indices of hepatic reserve should be reviewed. Indices recently attracting interest in liver resection cases are quantitative 99mTC-galactosyl human serum albumin scintigraphy using liver cell surface asialoglycoprotein receptor, and functional hepatic resection rate using 99mTC-GSA SPECT images, which is important in estimating liver regeneration after percutaneous trashepatic portal embolization. PMID- 9330376 TI - Trimethyltin syndrome as a hippocampal degeneration model: temporal changes and neurochemical features of seizure susceptibility and learning impairment. AB - The effects of trimethyltin on the hippocampus were investigated in terms of changes in histology, depth electroencephalography, learning acquisition and memory retention, choline acetyltransferase and neuropeptides, and seizure induced c-fos messenger RNA expression. The results were as follows. (1) Morphologically, trimethyltin produced a progressive loss of hippocampal CA3 and CA4 pyramidal cells, starting from four days after peroral treatment with trimethyltin hydroxide (9 mg/kg), as described previously. (2) Neurophysiologically, the increased seizure susceptibility to pentylenetetrazol treatment reached a maximum at four days post-trimethyltin and then declined after five days post-trimethyltin. The maximal seizure susceptibility at four days post-trimethyltin was confirmed by the immediate and long-lasting appearance of spike discharge in the hippocampus. However, this was not verified by the expression of c-fos messenger RNA in the hippocampus, which was comparable between trimethyltin-treated and control rats. (3) Behaviorally, the time-courses of aggression and learning impairment were similar to that of the seizure susceptibility. (4) Neurochemically, trimethyltin treatment caused changes of neurochemical markers, which were manifested by the elevation of neuropeptide Y content in the entorhinal cortex, and of choline acetyltransferase in the hippocampal CA3 subfield. Trimethyltin may offer potential as a tool for investigations on the relationship between neuronal death in the hippocampus and the development of seizure susceptibility and learning impairment. Alterations in glucocorticoids, glutamate and neuropeptides may all contribute to the manifestation of the trimethyltin syndrome. PMID- 9330378 TI - [Perioperative management for patients with chronic liver injury and the strategy for preventing postoperative liver failure]. AB - In liver surgery, postoperative liver failure has been a serious problem of concern. Recently, the advances in the imaging diagnosis and in the operative procedures have contributed to reduce the operative mortality to less than 1%. Between October 1994 and December 1996, a total of 159 patients, including 39 with chronic hepatitis and 66 with cirrhosis, underwent liver resection for hepatocellular carcinomas (n = 103), metastatic tumors (n = 24) and others (n = 32). Although about 20% of patients had some postoperative complications, no patient died of postoperative liver failure. Preoperatively, the liver function was estimated by ICG R15 and CT volume metry, and portal vein embolization and the splenectomy, if necessary, was performed. Blood loss was replaced by plasma as far as possible. Postoperatively, it is most important to maintain the optimal water balance and electrolyte levels using fresh plasma and diuretics. Even in patients with cirrhosis, no operative mortality can be achieved with optimal hepatectomy and careful perioperative management. PMID- 9330379 TI - [Thoracic surgery and liver dysfunction]. AB - Lung resection results in loss of lung parenchyma including residual healthy lung tissue and in reduction in pulmonary vascular bed. A decrease in residual pulmonary vascular bed after lung resection causes an increase in right heart afterload, and in some patients, it would be associated with an increase in right heart preload and consequent the changes in hepatic circulation which would lead to liver damage. Preceding thoracotomy, unilateral pulmonary arterial occlusion test (UPAO) was performed to simulate the hemodynamic changes after lung resection to evaluate the increase in right heart preload after surgery. Patients with the decreases in cardiac index or PaO2 during UPAO showed a higher levels of GPT during postoperative period when compared with those with the increase in either parameters. In a surgical treatment for empyema, bronchiectasis, or other infectious lung diseases, bronchial angiography (BAG) and also bronchial arterial embolization (BAE) were useful methods to prevent from exceeding bleeding during thoracotomy, which is one of the risk factors to cause liver damage after surgery. These results suggest that, in the field of thoracic surgery, the preoperative assessment of the hemodynamic changes caused by lung resection and the preoperative attempt to prevent from bleeding during thoracotomy are both important to protect from liver damage caused by surgical stress. PMID- 9330380 TI - [Cardiovascular surgery in the patients with liver dysfunction]. AB - Under the current situations we have increasing opportunities to manage the patients with posthepatitis and/or congestive liver dysfunction. In order to prevent postoperative hepatic failure we described perioperative management for those patients. For this purpose, the major point is the selection of appropriate operative methods which reduce operative invasions and have sure efficacy. In order to decide operative methods, we have to grasp the functional reserve of dysfunctional liver. We have no effective methods to estimate the functional reserve, but our data suggested that serum cholinesterase level at the preoperative states might demonstrate prognostic significance. The others are managements of postoperative cardiopulmonary distress and infection. It goes without saying that preoperative improvement of anemia and poor nutrition. PMID- 9330381 TI - [Surgery for upper gastrointestinal diseases in cirrhotic patients]. AB - Outcomes of surgery for gastric cancer or esophageal cancer in cirrhotic patients are not favorable. The preoperative assessment of liver function utilizing Child's classification or indocyanine green (ICG) excretion test can be a predictive factor of postoperative mortality. Operative risk is acceptable if patients are classified as Child's class A, and surgical procedures should be avoided in patients either classified as Child's class C or having ICG-R15 of 25% or more. To avoid postoperative complications, it is important to minimize the operative procedure and to ligate vessels instead of using electrocautery. Surgical stress and risk can further be reduced by a two stage operation for esophageal cancer and by gastrectomy with reduced lymph node dissection of D1 for gastric cancer. However, because curability of existing cancer is also required for surgical procedures, the status of liver cirrhosis and the stage of cancers should be considered in surgical treatment of gastric cancer or esophageal cancer in patients with liver cirrhosis. PMID- 9330382 TI - [Surgical treatment for colorectal carcinoma combined with liver cirrhosis]. AB - Surgical treatment for colorectal carcinoma associated with liver cirrhosis is discussed. Resection of this carcinoma is considered safe for patients whose liver function belongs to clinical stage I, II or Grade A, B of Child-Pugh classification or KICG more than 0.06/min. When hepatocellular carcinoma (HCC) is associated and the remnant liver function is feasible, hepatic resection should be undertaken at a favorable opportunity. When risky esophageal varices are present, endoscopic variceal ligation (EVL) or sclerotherapy (EIS) should be performed preoperatively. Lymph node dissection should be restricted to D1 or D2. In the postoperative phase, attention should be paid to maintaining liver function and hepatic circulation. Dobutamine or dopamine is useful for increasing hepatic blood flow. Fresh frozen plasma or platelet transfusion is used when needed. PMID- 9330383 TI - [Surgery for cholelithiasis in cirrhotic patients]. AB - Although biliary tract surgery for cholelithiasis is performed frequently, cirrhotic patients require special consideration. The prevalence of postoperative severe complications, such as hepatic failure and biliary peritonitis caused by insufficient fistula formation after removal of the T-tube, is higher than non cirrhotic patients. We suggest that definitive surgery can be carried out safely, in Child's A and B cirrhotic patients, either electively or as an emergency. However, a more conservative approach is advisable in Child's C patients with acute conditions and definitive surgery is recommended as an elective procedure after liver function has improved. And for the treatment of choledocholithiasis in patients with severe cirrhosis, avoiding surgical intervention through the use of such techniques as endoscopic papillotomy is recommended whenever possible. PMID- 9330384 TI - [Surgical therapy for hepatocellular carcinoma with liver cirrhosis]. AB - Approximately 80% of hepatocellular carcinoma (HCC) patients in Japan have associated liver cirrhosis, which increases the difficulty of surgical treatment. Liver dysfunction associated with liver cirrhosis is one of the most important predictive prognostic factors for HCC patients. Percutaneous ethanol injection therapy (PEIT) is useful for patients with small HCC or with poor hepatic functional reserve. Transcatheter arterial chemoembolization (TACE) is also useful both for patients with unresectable HCC and patients with multiple intrahepatic recurrence. Liver resection, however, lead to better outcome than other treatments when liver function is maintained after surgery. To determine operative procedures, it is important to evaluate the exact function of remnant liver, based on the preoperative liver function test and the evaluation of tumor character. For advanced HCC patients with vascular invasion, non-surgical treatments such as PEIT or TACE are not indicated, and surgical intervention can be an effective modality to improve their survival. Improvements of surgical technique and perioperative management have decreased fatal complications at a major liver resection and allowed us to carry out liver resection on patients with advanced HCC. PMID- 9330385 TI - [The significance of the medical research study: hemodynamic and pathophysiologic characteristics in peripheral arterial reconstruction]. AB - In cases of peripheral arterial reconstructive surgery, it is important to prevent postoperative early and late occlusion of the reconstructed artery. The postoperative early outcome of the reconstructed artery were related to electromagnetically determined flow waveform of the reconstructed artery. Flow waveform was classified in 5 types, i. e. types O, I, II, III, and IV. The prognosis of the reconstructed arteries with types O and I was excellent. Type II showed no postoperative early occlusion but late occlusion did occur in 31%. Type III and IV were all failed within 48 hours postoperatively. Correlation between prognosis of the reconstructed artery and flow waveform was established by the intraluminal velocity profile of blood flow, using a specially designed flow wave simulation pump system. Postoperative late occlusion in arterial reconstruction often occurs in cases with poor distal runoff and caused by intimal hyperplasia of implanted autovein graft and its distal end-to-side anastomosis. The features of intimal hyperplasia of the implanted autovein graft differed from those at the distal end-to-side anastomosis; the former being related to active proliferation of smooth muscle cells while the latter displayed an excessive proliferation of fibroblasts and collagen fibers. PMID- 9330386 TI - [Results of surgical treatment for thoracic esophageal cancer with reference to a randomized controlled trial]. AB - An esophagectomy with lymphadenectomy was performed on 248 patients in The Second Department of Surgery, Tohoku University Hospital from 1986 to 1992. Five year survival rate of 211 patients who underwent curative operation (RO) was 53.1%, however that of 33 patients undergone non-curative operation (R1 and R2) was 8.0%. The operative death was only one (0.4%) in this series. In order to clarify the efficacy of the extensive lymphadenectomy, a randomized controlled trial was performed during the same period. Five year survival rate of extensive lymphadenectomy group (n = 23) was 61.1%, however that of conventional lymphadenectomy group (n = 22) was 48.4%. There were some metastatic nodes in the left upper mediastinum or in the neck where could not be dissected through the conventional lymphadenectomy. These results indicate that the extensive lymphadenectomy for thoracic esophageal cancer can be safely performed and may contribute to improvement of survival rate after surgery. PMID- 9330387 TI - [New aspects in the liver regeneration following partial hepatectomy]. AB - We demonstrated the new two concepts for the liver regeneration after partial hepatectomy (PHx). The first is immunological findings in the regenerating liver. Th1 type T cells, which produce IFN gamma, and extrathymic T cells may play an important role in immunological control system of liver regeneration after PHx on the basis of the cellular immunity restricted by MHC class I and II molecules. The second is the mechanism of "on and off" on liver regeneration after PHx. Portal pressure, which reflecting wall shear stress of sinusoid, may trigger the liver regeneration and control the liver volume after PHx. There are two type of intrahepatic leukocytes; one type would tend to stay associated with SEC, while the other would not. Therefore shear stress may have a great influence on the leukocytes-sinusoidal endothelial cells (SEC) adhesion immediately after partial hepatectomy. PMID- 9330388 TI - [A case of sudden onset of toxic cholangitis]. AB - A previously healthy 43-year-old male was admitted to the hospital because of the impared consciousness. Although a passage of the bile duct was good, the patient was in severe biliary sepsis and cardiac arrest occurred repeatedly. Plasma concentration of TNF increased markedly suggesting a pathogenic role of TNF in the course of the patient. Plasma exchange was effective for hemodynamic stabilization. Recent advances in treatment of acute cholangitis has been brought by early drainage of the obstruction in the bile duct using endoscopic or percutaneous drainage. However this case suggests that only drainage of the infection focus is not sufficient for the life saving in such a patient and other therapeutic approach such as an inactivation of TNF activity should be considered. PMID- 9330389 TI - [Molecular changes during differentiation in glioma cells]. PMID- 9330390 TI - [Neuroendoscopic surgery for hydrocephalus]. PMID- 9330391 TI - [The effect of Japanese herbal medicine on MRSA carrier in neurosurgery]. AB - Since the management of methicillin-resistant staphylococcus aureus (MRSA) carriers is not established, these patients are isolated individually, limited in activity, and delayed in rehabilitation. In this study, the effect of Japanese herbal medicine on MRSA carriers was examined. In the control group, MRSA carriers were isolated individually. In the treatment group, one of the Japanese herbal medicines "Juzentaihotou" or "Hotyuekkitou" was given in addition to isolating the patient. It was shown in cultures that Japanese herbal medicines had effectively changed MRSA carriers to negative. They also shortened the duration required to bring about the change of MRSA carriers to negative. As a result, the total number of MRSA carriers was reduced. These results suggested that Japanese herbal medicines may be useful for the management of MRSA carriers in neurosurgery. PMID- 9330392 TI - [Acute subdural hematoma due to pure cortical arterial injury: analysis of 14 cases]. AB - Pure cortical arterial injury is defined as an acute subdural hematoma (ASDH) due to micro laceration of the cortical artery without cerebral contusion. We analyzed this peculiar ASDH in 14 cases. This ASDH has the following characteristics. 1) This disease develops in elderly persons after minor head trauma. 2) After a relatively long lucid interval, the consciousness level of these patients decreases rapidly to semicoma or deep coma. 3) Brain atrophy, atherosclerosis and a tendency to bleed (drugs, hemodialysis, thrombocytopenia, etc.) are involved in this hematoma. 4) The clinical outcome is generally poor because of systemic complications. The authors infer the mechanism of pure cortical arterial injury to be as follows. Minor hemorrhage occurs due to direct impact damage after trivial head trauma, but physiological hemostatic actions are set in motion. After a lucid interval, rebleeding from the injured cortical artery causes massive subdural hemorrhage. PMID- 9330393 TI - [Management of subarachnoid hemorrhages without detectable aneurysm]. AB - In this study, 21 patients with subarachnoid hemorrhage (SAH) but negative angiography were evaluated. Angiography was performed twice on each patient, that is, on admission and at 2 weeks following admission. All patients had severe headache of sudden onset, a characteristic manifestation of SAH. Clinical grades on admission (Hunt and Kosnik classification) were generally good: 17 patients were in grade I or II and 4 patients were in grade III. SAH was confirmed by the presence of subarachnoid clot on CT in all cases. Based on the distribution of SAH, CT findings were classified into two patterns, i.e., perimesencephalic and non-perimesencephalic patterns. Four patients showed the perimesencephalic pattern and the remaining 17 the non-perimesencephalic. The period of follow-up ranged from 20 days to 11 years 6 months, with a mean of 6 years 10 months. Except for three recent cases, the mean follow-up period in 8 years 9 months. Exploratory craniotomies probing for aneurysms have been performed in four patients, but no aneurysms have been found in any of these cases. Clinical deterioration associated with vasospasm was observed in one patient. A communicating hydrocephalus requiring a shunting procedure was observed in three patients showing the non-perimesencephalic type CT pattern. Rebleeding occurred in one patient who subsequently died of what may be a dissecting aneurysm of the vertebral artery. One patient who was able to return to full activity experienced symptoms attributable to SAH such as frequent headaches and increased fatigability. Complete recovery was observed in the remaining 19 patients. Two of them, however, later died due to myocardial infarction and aging, respectively. Given these generally positive outcomes, it should be possible to inform such patients of the benignity of their condition. Angiography may not demonstrate a ruptured aneurysm on initial examination in all cases of aneurysmal SAH. Serial angiography, however, can provide a definite diagnosis of the dissecting aneurysm. Therefore, repeat angiography, particularly, when possible, digital subtraction angiography, is necessary to rule out aneurysmal SAH. While small aneurysms or microaneurysms are often found through exploratory craniotomy, we do not agree with the opinion that surgery may be appropriate for certain patients with SAH but with negative angiography. The natural history concerning rebleeding in such cases, as well as morbidity and mortality associated with hemorrhage, remains to be defined. Furthermore, there are reservations regarding whether coagulation of these abnormalities with bipolar cautery constitutes definitive treatment. PMID- 9330394 TI - [Usefulness of fronto-orbito-zygomatic approach for intraorbital tumors: report of 31 cases]. AB - We have encountered 31 cases of intraorbital tumors at the Kobe University Hospital between 1971 and 1996. We reviewed those cases taking into account the surgical approaches used, the removability of the tumors and functional outcome. Before 1991, frontal or fronto-temporal craniotomy had been mostly used whereas the fronto-orbito-zygomatic approach or fronto-orbital approach has been used only since 1992. After introduction of these approaches, functional outcome was significantly improved. Most of the tumors have been successfully removed totally. In particular, this approach is thought to be useful for tumors located in the deep portion of the orbit and for tumors extending intracranially. In addition, removal of the anterior clinoid process and the opening of the optic canal as well as the superior orbital fissure provides a wider operative view and enables the transposition of the optic nerve safely. PMID- 9330395 TI - [Stereotactic brachytherapy for clival chordoma]. AB - There has recently been interest in the use of high-dose radiation with methods such as radiosurgery and brachytherapy for skull base tumors. Brachytherapy is believed to be effective for clival chordomas, but technical difficulties exist in stereotactic insertion of catheters into the clivus. We assessed the usefulness following improvement of implantation techniques in three patients with clival chordomas. All tumors were larger than 50 mm in diameter. Removable iridium-192 sources were held in catheters which were implanted through a transnasal approach under general anesthesia using a CT-guided stereotactic system in one patient and a CRW stereotactic system adapted to a magnetic resonance imaging (MRI) scanner in 2 patients. The implantation array was designed based on results of stereotactic 3-D MRI scanning, and coordinates were calculated for stereotactic implantation through twist drill holes. These catheters were introduced through the nares and directed into the clival chordoma under endoscopic visualization and X-ray fluoroscopy. No complications such as CSF liquorrhea, hemorrhage or infection were observed. Brachytherapy with a total dose of 43.2-58.0 Gy at the tumor periphery was administered for 7 to 10 days, and serial follow-up imaging studies demonstrated reduction in tumor size in two patients and no tumor growth in the other. Our results suggested that stereotactic brachytherapy is potentially useful for the control of clival chordomas and that computer-guided transnasal stereotactic insertion enables implantation of catheters less invasively and more accurately than does X-ray fluoroscopic guidance alone. PMID- 9330396 TI - [Objective evaluation of selective peripheral denervation for spasmodic torticollis]. AB - The authors evaluated the results of selective peripheral denervation (SPD) of posterior rami of the cervical spinal nerves and/or the accessory nerve for spasmodic torticollis. Five patients underwent seven operations in total and the results were evaluated with the modified Tsui's score which was used in the clinical trial of botulinum toxin (BTX) for torticollis in Japan. The preoperative score was 10.8 +/- 2.2 (mean +/- S.D.) and the postoperative score was 1.4 +/- 1.7. The score changes indicated the effects of the operation as "excellent" in four cases and "good" in one case. These results indicate that SPD is superior to BTX in terms of control of symptoms in spasmodic torticollis. After the initial operation, however, two patients showed the so-called "mole hitting game phenomenon" in which normal muscles develop abnormal contraction after denervation of abnormal muscles. This forced us to perform the second operations. Although this phenomenon was first described in botulinum toxin treatment, this is probably the first report in surgically denervated cases. PMID- 9330397 TI - [An unbranched-site ruptured aneurysm of callosomarginal artery]. AB - A case of ruptured callosomarginal artery aneurysm was reported. A 47-year-old male was admitted to our hospital with disturbance of consciousness. CT scan revealed subarachnoid hemorrhage. No aneurysm was detected by initial angiography. His consciousness level deteriorated so we stopped the examination. CT scan after initial angiography showed intracerebral hematoma in the left frontal lobe. On day 15 severe vasospasm occurred and he presented with total aphasia and right hemiplegia. The third angiography disclosed a callosomarginal artery aneurysm. Neck clipping of the aneurysm was successfully performed via unilateral interhemispheric approach. Postoperative course was uneventful and the patient recovered well with mild right hemiparesis and motor aphasia. He was transferred to another hospital for rehabilitation. The incidence of distal anterior cerebral artery aneurysm has been reported as being about 5%. The location of aneurysm in this case is very rare. In the present study we review the literature on aneurysms at the distal anterior cerebral artery and discuss the clinical and radiological features. PMID- 9330398 TI - [A case with nasopharyngeal carcinoma extending into the cavernous sinus]. AB - Nasopharyngeal carcinoma is seen frequently in South China, but it is rare in Japan as well as in Western countries. We reported a rare case with intracranial extension at the initial presentation. This 25-year-old Japanese female showed left abducens palsy. MRI revealed an intracavernous mass with homogeneous enhancement connected with the lesion around the epipharynx through the parapharyngeal space. The mass enlarged gradually, followed by left trigeminal palsy. An elevation of the titer of serum EBV Ig-G was noted. She was operated on via the subtemporal approach, which revealed an intracavernous tumor, connected with the infratemporal fossa mass via the enlarged foramen ovale. Histological examination of the surgical specimen revealed lymphoepithelioma. Local irradiation of 60Co of 63 Gy was performed on the cavernous and parapharyngeal tumors and they decreased in size remarkably. However, metastasis to the upper right neck lymph node occurred, and it was removed. Additional irradiation of 45 Gy was performed on her neck. Chemotherapy was not able to be carried out due to leucocytopenia. At present, 2 years after onset, no regrowth of the tumor has been seen. At initial presentation, intracranial extension of nasopharyngeal carcinoma is very rare. Remission may be obtained by a combination of surgery and irradiation even in cases with metastasis. PMID- 9330399 TI - [A case of epidermoid tumor inside the Meckel's cave]. AB - An epidermoid tumor inside the Meckel's cave is rare. The symptoms caused by this tumor include trigeminal neuralgia, facial hypesthesia and paresis of the 3rd, 4th and 6th nerves. A case of epidermoid tumor inside Meckel's cave was presented. A 54-year-old female who had complained of 3rd nerve palsy with right facial hypesthesia since 3 years before was referred to our clinic. Magnetic resonance imaging (MRI) showed the tumor at Meckel's cave. The tumor removal was performed using the orbito-zygomatic approach. To avoid injury of the internal carotid artery and nerves inside the cavernous sinus, removal of the tumor inside the capsule was carried out leaving the capsule. Postoperatively, the tumor removal was confirmed by MRI and improvement of the 3rd and the 5th nerve palsy was obtained three months after surgery. This case suggests that the capsule of the tumor inside the Meckel's cave should be allowed to remain to avoid injury of the adjacent 4th, 5th and 6th nerves and of the internal carotid artery. PMID- 9330400 TI - [Subependymal tumor with metaplastic bone formation: a case report]. AB - A 26-year-old woman was admitted to our hospital because of headache. CT scan and MRI showed a right subependymal nodule and a left ventricular tumor, neither of which had any enhancement nor were they stained in angiography. Although no skin abnormality was detected, the patient was suspected of tuberous sclerosis. The diagnosis was made because of the subependymal nodule on CT scan and MRI. On June 29, 1995, total removal of a left ventricular tumor was performed by a transcortical approach. Histological sections of this tumor consisted of astrocytic and meningothelial components, containing metaplastic bone formation. Histological diagnosis was dysplastic subependymal tumor. Postoperative course was uneventful. Regrowth of the tumor has not been observed as of now. This case was suspected to involve factors of tuberous sclerosis from a subependymal nodule. However, the ventricular tumor was not diagnosed as a subependymal giant cell astrocytoma. PMID- 9330401 TI - [Diffuse cerebral artery vasospasm following total resection of posterior fossa meningioma: a case report]. AB - Diffuse cerebral artery vasospasm following brain tumor resection is a rare complication. The authors reported a case of symptomatic diffuse cerebral artery vasospasm of early phase following resection of a left posterior fossa meningioma. A 50-year-old female patient was admitted to our hospital complaining of headache. No neurological deficits were detected at the time of admission. Computed tomography (CT) and magnetic resonance imaging (MRI) showed a large mass in the left posterior fossa. Cerebral angiography demonstrated mildly diffuse stenosis of the bilateral internal carotid artery. The tumor was resected totally. CT after operation showed a small amount of subarachnoid hematoma in the superior aspect of the cerebellum. Pathological specimen of the tumor showed fibrous meningioma. One day after this radical operation, the patient was found to have weakness in her left leg. Then she developed left hemiparesis, weakness in the right leg and left homonymous hemianopsia. MRI showed ischemic lesions in the bilateral parietal and the occipital lobe. Angiography demonstrated diffuse severe vasospasm throughout the whole cerebral artery. Ten days after the operation, angiographical findings were improved. This case indicates that vasospasm may occur even after resection of brain tumors which are localized outside the suprasellar area. PMID- 9330402 TI - [A case of tuberous sclerosis presenting intractable adversive seizure, successfully resected with the technique of "gyrectomy"]. AB - We report a 9-year-old girl with tuberous sclerosis presenting intractable adversive seizure. She had been suffering from frequent attacks of consciousness loss since the age of 6 years. Although a considerable amount of antiepileptic drugs had been administered, her epileptic attacks were not controlled, but instead rather increased. She had been suffering from adversive seizure to the right for more than 2 years. CT scan failed to show any abnormal density area. MRI showed a small lesion in the left frontal subcortical area. The electroencephalogram showed relatively mild abnormal waves in the left hemisphere. We undertook surgical removal of the lesion with epileptogenic foci because her epilepsy has not been controlled and the lesion could be a glioma. Abnormal spike waves were detected around the lesion with electrocorticogram. "Gyrectomy" technique was employed and the spike waves totally disappeared. After the surgery, no neurological deterioration was presented. She has suffered no seizure attack since the surgery even though the amount of the antiepileptic drugs has been significantly decreased. Resection of the epileptogenic foci as well as the abnormal lesion using the technique of gyrectomy is useful for the control of the intractable epilepsy, and makes the quality of life of patients much higher. PMID- 9330403 TI - Management of locally advanced breast cancer. AB - Multimodality therapy--i.e., surgical excision followed by appropriate systemic therapy and radiotherapy--has an established role in managing patients with locally advanced breast cancer (LABC). Preoperative chemotherapy permits optimal local control with less radical surgical intervention, although its impact on overall survival is still unclear. Definitive data are not yet available to determine the optimal sequencing of surgery and radiation therapy. Therefore, treatment should continue to be individualized. New cytotoxic agents with demonstrated activity against metastatic breast cancer (e.g., the taxanes) are being studied to determine their role in women with LABC. Preliminary data from a recently completed, small randomized trial in patients with LABC did not demonstrate a significant improvement in overall survival with high-dose chemotherapy plus stem-cell rescue, as compared with standard-dose therapy. The evaluation of biologic parameters that may predict response and survival, and of radiographic and pathologic methods to assess response, should ultimately lead to significant improvements in the management and survival of patients with locally advanced breast cancer. PMID- 9330404 TI - Combined-modality therapy for bladder cancer. AB - Radical cystectomy remains standard management for patients with locally advanced T2 through T4, N0, M0 transitional cell carcinoma of the urinary bladder. Although radical cystectomy results in excellent local control, 50% or more of patients relapse. Studies have shown that multidrug cisplatin (Platinol)-based chemotherapy prolongs disease-free survival in 10% to 15% of cases and is superior to single-agent cisplatin. These studies led to the application of these regimens in conjunction with surgery and/or radiation therapy in an attempt to preserve bladder function. With this approach, the decision to leave the bladder in place, remove a portion (partial cystectomy), or perform a radical cystectomy is made after assessing the initial response to therapy. Results from neoadjuvant studies have shown that: major responses are observed in at least 50% of patients; bladder preservation can be achieved in 25% to 50%; a pathologic complete response predicts long survival; and patients with deeply invasive lesions (T3b to T4) usually are not candidates for bladder preservation. Whether overall survival is improved has been difficult to ascertain due to such issues as small sample size and case selection. Concurrently, newer surgical approaches with continent diversions have reduced, to some extent, the need for ileal conduits, a factor influencing the bladder preservation approach. Adjuvant chemotherapy, although less well studied, suggests a possible survival benefit for selected patients with a high likelihood of relapse. To optimize patients selection, new prognostic factors are necessary. Many biologic variables based on expression of tumor-related proteins are under study. Combined-modality therapy is not standard management for the majority of bladder-cancer patients. However, it is a viable alternative for those who are committed to preserving bladder function. Additional research is required to determine whether these approaches improve survival and to identify better markers of treatment outcome. PMID- 9330405 TI - Combined-modality therapy of head and neck cancer. AB - The treatment of head and neck cancer has traditionally consisted of surgery with postoperative radiation therapy. Chemotherapy has been reserved for palliation. In recent years, induction chemotherapy has been shown to allow for larynx preservation in patients with laryngeal or hypopharyngeal cancer. Concomitant chemoradiotherapy has been shown to provide a statistically significant survival benefit in selected studies and recent meta-analyses. In patients with nasopharyngeal cancer, the use of cisplatin and concomitant radiation therapy results in a marked survival benefit. PMID- 9330406 TI - Concurrent chemoradiotherapy for limited small-cell lung cancer. AB - It is now established that the treatment of choice for limited small-cell lung cancer (SCLC) in the United States, Canada, and Japan is thoracic radiotherapy (TRT) combined with etoposide (VePesid), either alone or in conjunction with other agents, especially a platinum agent. The specific factors related to the use of TRT in the treatment of limited SCLC are: (1) dose (total and daily), (2) volume to be irradiated, (3) fractionation, (4) timing of radiation relative to chemotherapy (concurrent, at the same time; alternating, using both within weeks; or sequential, all of one followed by all of the other without any overlap), (5) whether radiation should be given earlier or later in the treatment course, and (6) whether to use a split course (rest intervals during a course of radiotherapy) or a continuous course of radiation. This paper discusses each of these factors. PMID- 9330407 TI - New drugs for small-cell lung cancer. AB - Better efficacy of chemotherapy for small-cell lung cancer (SCLC) has not been convincingly demonstrated despite many investigations examining increased drug delivery and enhancement of drug diversity. Improvements in the median- and long term survival of patients with limited SCLC have been shown, but this change is more closely associated with better integration of chemoradiation than with more effective chemotherapy or radiotherapy components. Recently, a number of new active drugs have been identified for treating patients with SCLC. These drugs include the taxanes, the camptothecins, and gemcitabine. Determining the role of each new agent will require carefully designed and conducted trials like those now in progress. PMID- 9330408 TI - Combined-modality therapy of locally advanced non-small-cell lung cancer. AB - Treatment of patients with unresectable stage IIIA and IIIB non-small-cell lung cancer with conventionally-fractionated radiation therapy (i.e., total doses of 50 to 60 Gy, using one fraction per day), which was standard practice in the 1970s and early 1980s, resulted in good short-term palliation but few long-term survivors. Local control was poor, and the majority of patients also rapidly developed symptomatic metastatic disease outside the chest. In the past 15 years, a number of approaches to improve this situation have been defined in prospective clinical trials. They include radiation therapy with altered fractionation schemes that allow either higher overall doses or shortened treatment times, the use of systemic chemotherapy to address microscopic metastatic disease, and the use of a variety of agents, some but not all with intrinsic cytotoxic activity, to act as radiation sensitizers. These strategies have resulted in modest but significant improvements in local and systemic disease control, but at a cost of increased toxicity, including myelosuppression, esophagitis, and pneumonitis. Further advances in treatment will require better (i.e., more active) cytotoxic agents and better ways of limiting radiation effects to the target volume of tumor. PMID- 9330409 TI - Induction chemotherapy with/without radiation followed by surgery in stage III non-small-cell lung cancer. AB - The objectives of this review are to provide an update and perspectives on the use of induction therapy (chemotherapy with or without radiotherapy) followed by surgery in two subgroups of patients with stage III non-small-cell lung cancer. The first subset is that of bulky stage IIIAN2 or IIIB disease (standard treatment: chemoradiotherapy), and the second, minimal stage IIIA non-N2 or computed tomography (CT)-negative N2 disease (standard therapy: initial surgical resection). Details of recent major trials in each of these two subsets are provided regarding selections criteria, study design, toxicity, resection rates, median and long-term survival, and predictors of survival. The review concludes with a discussion of whether consensus has emerged about the addition of surgery after induction chemoradiotherapy in the group with bulky disease and whether there should now be a standard recommendation for preoperative chemotherapy with or without radiotherapy in patients with initially resectable tumors. PMID- 9330410 TI - Precise clinical staging allows treatment modification of patients with esophageal carcinoma. AB - Treatment of esophageal carcinoma requires the realization that this neoplasm is not a single entity with a uniformly poor prognosis. As with any other malignancy, disease stage has prognostic and therapeutic importance. Patients with stage T1-2,N0,M0 or lower tumors have acceptable surgical cure rates and should undergo immediate resection. Patients with more advanced tumors (T3 or N1) are still potentially curable but do poorly with surgery alone. These patients should be considered for multimodality therapy. Patients with hematogenous metastatic disease should be treated with palliative intent. Endoscopic ultrasound (EUS), an accurate, reproducible staging tool, allows for clinical staging of these patients and modification of treatment at the time of diagnosis. Our experience at The Cleveland Clinic Foundation using this algorithm of surgery alone in patients with EUS-defined early-stage carcinomas and multimodality therapy in patients with EUS-staged locally advanced disease is described. PMID- 9330411 TI - Combined-modality treatment of esophageal cancer. AB - The use of chemotherapy and radiotherapy prior to surgery for patients with potentially resectable esophageal carcinoma has been investigated since the late 1970s, with trials yielding response rates approaching 50%. Although some of these trials suggested improved survival with preoperative chemoradiation plus surgery, as compared with historical controls treated with surgery or radiation alone, the question of survival benefit had to be addressed in prospective, randomized fashion. This review describes the experience with surgery or radiation alone vs preoperative chemoradiation in terms of long-term overall survival in the treatment of esophageal cancer. PMID- 9330412 TI - Ventilatory response of the newborn infant to mild hypoxia. AB - The transition from an immature (biphasic) to a mature (sustained hyperpneic) response to a brief period of sustained hypoxia is believed to be well advanced by postnatal day 10 for newborn infants. However, a review of the supporting evidence convinced us that this issue warranted further, more systematic investigation. Seven healthy term infants aged 2 days to 8 weeks were studied. The ventilatory response (VR) elicited by 5 min breathing of 15% O2 was measured during quiet sleep. Arterial SaO2 (pulse oximeter) and minute ventilation (expressed as a change from control, delta V'i) were measured continuously. Infants were wrapped in their usual bedding and slept in open cots at room temperature (23 degrees-25 degrees). Infants aged 2-3 days exhibited predominantely a sustained hypopnea during the period of hypoxia (delta V'i = -2% at 1 min, -13% at 5 min). At 8 weeks of age, the mean response was typically biphasic (delta V'i = +9% at 1 min, -4% at 5 min). This age-related difference between responses was statistically significant (two-way ANOVA by time and age group; interaction P < 0.05). These data reveal that term infants studied under ambient conditions during defined quiet sleep may exhibit an immature VR to mild, sustained hypoxia for at least 2 months after birth. This suggests that postnatal development of the O2 chemoreflex is slower than previously thought. PMID- 9330413 TI - Decreased concentration of exhaled nitric oxide (NO) in patients with cystic fibrosis. AB - Nitric oxide (NO) is produced by various cell types in the human respiratory tract. Endogenously produced nitric oxide is detectable in the exhaled air of healthy individuals. Exhaled NO has been shown to be increased in airway inflammation, most probably due to cytokine-mediated activation of NO synthases. To assess whether NO can serve as a marker of inflammation in cystic fibrosis (CF) lung disease, we measured exhaled NO in CF patients with a chemiluminescence analyser. Single breath measurements were performed in 27 stable CF patients (age range, 6-40 years) and 30 non-smoking controls (age range, 6-37 years). Exhaled NO concentrations were 9.1 +/- 3.6 ppb in the controls and 5.9 +/- 2.6 ppb (P < 0.001) in CF patients. To account for room air NO concentrations on the measurement of exhaled NO, we also calculated the difference between exhaled NO and ambient NO concentrations. Difference values were also significantly lower in CF compared with controls (P < 0.0001). In CF patients there was a positive correlation between exhaled NO and forced vital capacity (r = 0.43, P = 0.033), suggesting that exhaled NO is lower in patients with severe lung disease than in those with mild disease. We conclude that measurements of exhaled NO in CF does not reflect activity of CF airway inflammation. The decreased concentrations of exhaled NO may be due to inhibitory effects of inflammatory cytokines on NO syntheses in the airways and alveolar epithelial cells or to increased retention in airway secretions. PMID- 9330414 TI - Growth during one year of treatment with fluticasone propionate or sodium cromoglycate in children with asthma. AB - The aim of this study was to compare accurately measured growth over 12 months in asthmatic children treated with either fluticasone propionate (FP) 50 micrograms twice daily (b.i.d.) or sodium cromoglycate (SCG) 20 mg four times daily (q.i.d.). After a 2-week run-in, asthmatic children aged 4-10 years from 15 UK centers were randomized in a 3:4 ratio to open-label FP (n = 52) or SCG (n = 70). After 8 weeks, those whose asthma was not adequately controlled were switched from SCG to FP or withdrawn. Standing height was measured (Holtain stadiometry) at baseline, after 8 weeks and at 6 weeks intervals thereafter for 1 year. Morning peak flows (PEFam) were recorded by patients for 2 weeks during baseline, and 1 week before each visit during treatment. Urinary free cortisol (24 h) was measured at baseline, 6 months, and 1 year. After 8 weeks, 22 patients were withdrawn from SCG group (and were switched to FP), and five patients were withdrawn from the FP group due to poor asthma control. A further 21 and 11 patients were withdrawn from the SCG and FP groups, respectively, during the course of the study. There were no significant differences between patients who received FP and SCG for 1 year (n = 34 and n = 26, respectively) in terms of height velocity adjusted for age and gender (HV), or height velocity standard deviation scores adjusted for gender (HVSDS). Mean HV (mean HVSDS) were 6.0 cm/yr (0.1) and 6.5 cm/yr (0.5) for FP and SCG, respectively. There were no treatment differences in mean 24 h urinary free cortisol levels at 6 and 12 months. Mean % predicted PEFam improved over 1 year in both groups but to a greater degree in the FP group. We concluded that growth was normal in mildly asthmatic children receiving FP (50 micrograms bid) for 1 year. There were fewer withdrawals and lung function improved to a greater extent in FP treated patients than in patients receiving SCG. PMID- 9330415 TI - Effects of body fat on ventilatory function in children and adolescents: cross sectional findings from a random population sample of school children. AB - Childhood obesity is associated with a range of adverse consequences, and the prevalence is increasing in developed nations. Most of the literature on obesity and ventilatory function in children concerns samples selected for gross obesity with relatively little detail available from random population samples. This report examines the effect of total body fat as a percentage of weight (TBF%) on ventilatory function in a nationally representative sample of 2,464 Australian school children aged 9, 12, and 15 years. Forced vital capacity (FVC) and forced expiratory volume in 1 s (FEV1) were used as measures of ventilatory function. TBF% was estimated from skinfold thickness measurements. Ventilatory function was adjusted (for height and then for both height and weight) using linear regression on a logarithmic scale. Adjustment was performed within separate strata of age and gender. Analysis of covariance was used for hypothesis testing. Height adjusted FVC and FEV1 values increased significantly with increasing weight within each age and gender group and for all subjects combined (P < 0.0001). The effect of TBF% independent of lean tissue was examined using FVC and FEV1 values adjusted for both height and weight, because body weight measures both lean and fat mass. Adjusted FVC and FEV1 values decreased significantly with increasing TBF% within each age and gender group and for all subjects combined (P < 0.0001). Ventilatory function decreased with increasing proportions of body fat. This is consistent with previous findings on lean tissue mass and ventilatory function. Although the magnitude of the effect was relatively small in clinical terms, from a public health perspective our findings indicate yet another adverse consequence of childhood obesity. PMID- 9330416 TI - Increased incidence of sighs (augmented inspiratory efforts) during synchronized intermittent mandatory ventilation (SIMV) in preterm neonates. AB - A reflex resulting in a deep, sigh-like inspiratory effort (augmented breath) is frequently triggered during synchronized mechanical ventilation in preterm infants. We studied the incidence of augmented inspiratory efforts and their effect on ventilation and lung compliance during conventional IMV and synchronized IMV (SIMV) in 15 preterm neonates (GA 26.7 +/- 1.5 wks (mean +/- SD), BW 925 +/- 222 g, age 1-8 days). Augmentation of spontaneous inspiratory effort was defined as an esophageal pressure deflection occurring coincident with a synchronized mechanical breath and exceeding the previous unassisted spontaneous effort by more than 50%. The incidence of augmented breaths was higher during SIMV (11.1 +/- 7.7%; P < 0.01) than during conventional IMV (5.1 +/ 6.1%). However, when the synchronized breaths were triggered late (200-300 msec) after the onset of inspiration, augmented breaths occurred no more frequently than during conventional IMV (6.0 +/- 4.7%). The incidence of augmented breaths correlated inversely with dynamic lung compliance (P = 0.014), but was not significantly influenced by a change in PEEP. Although inspiratory effort increased nearly three times during the augmented breaths, tidal volume increased only 12%. The change in tidal volume was limited because the augmented effort reached its maximal negativity only approximately 500 ms after the beginning of the synchronized, mechanical breath and at a time when the mechanical breath had already ended. For this reason the augmented effort did not contribute significantly to minute ventilation, but only prolonged inspiration. Dynamic lung compliance did not change significantly after an augmented breath. The results indicate that augmented inspiratory efforts are more common in preterm neonates ventilated with SIMV than with conventional IMV, but do not contribute significantly to ventilation. PMID- 9330418 TI - Lung colonization with Enterobacteriaceae producing extended-spectrum beta lactamases in cystic fibrosis patients. PMID- 9330417 TI - Aerosol delivery to non-ventilated infants by metered dose inhaler: should a valved spacer be used? AB - In a randomized double-blind cross-over study on 20 spontaneously breathing, oxygen-dependent preterm infants who had received positive pressure ventilation for respiratory distress syndrome, we tested the hypothesis that the one-way non rebreathing valves of aerosol spacer devices might impair rather than enhance the delivery of aerosols to small infants by metered dose inhalers (MDI). Ten infants were given 2 doses (200 micrograms/dose) of MDI albuterol through a neonatal Aerochamber 4 h apart. At random sequence, one dose was delivered with the non rebreathing valve of the Aerochamber in place; for the other dose, the valve had been removed. The experiment was repeated on another ten infants using a different spacer device (Babyhaler) with or without its one-way inspiratory valve removed. During the first hour following aerosol administration, use of the non valved spacers was associated with a significantly greater degree of tachycardia in both groups, and also lower transcutaneous carbon dioxide tension in the Aerochamber group. All infants showed a reduction in respiratory system resistance and an improvement in functional residual capacity following albuterol treatment. In both groups, maximum reduction in respiratory system resistance, recorded 30 min after aerosol delivery, was significantly greater following the use of the non-valved spacers (Aerochamber: 51.2 +/- 3.1% vs. 35.0 +/- 2.8%, P < 0.0001; Babyhaler: 38.8 +/- 2.3% vs. 19.2 +/- 1.4%, P < 0.0001) than following the use of the spacers with a valve. The findings provide indirect evidence supporting our hypothesis and suggest that when the MDI is used to deliver therapeutic aerosols to non-ventilated newborns or small infants, a spacer device without a non-rebreathing valve should be used. PMID- 9330419 TI - Nasal CPAP treatment in an infant with respiratory syncytial virus-associated apnea. PMID- 9330420 TI - Surfactant aerosol treatment of respiratory distress syndrome in spontaneously breathing premature infants. PMID- 9330421 TI - Corticospinal tract regrowth. AB - The natural ability of the adult central nervous system of higher vertebrates to recover from injury is highly limited. This limitation is most likely due to an inhospitable environment and/or intrinsic incapacities of the neurons to re extend their neurites after injury or axotomy. The rat corticospinal tract is the largest tract leading from brain to spinal cord and is often used as a model in developmental and regeneration studies. The extensive know-how of factors involved in the development of the corticospinal tract did provide the foundation for many studies on corticospinal tract regrowth after injury in the adult spinal cord. The results of these experiments, as discussed in this review, have led to important contributions to the further understanding of central nervous system regeneration. PMID- 9330422 TI - Nerve growth factors and the control of neurotransmitter phenotype selection in the mammalian central nervous system. AB - Determination of neurotransmitter phenotype in the peripheral nervous system (PNS) has been intensively characterized. However, relatively little is known about the underlying molecular and biochemical events involved in determination of transmitter phenotype in the central nervous system (CNS). It has been well established that nerve growth factors regulate cell growth and differentiation. They are increasingly recognized as playing an important role in many decision making steps during development. Published data suggest that neurotransmitter phenotype is determined largely by exogenous stimuli, such as nerve growth factors--acidic/basic fibroblast growth factor, epidermal growth factor, neurotrophins, etc., working in concert with the genetic programmes. They exert potent effects independently or synergistically with other molecules by acting either on neural precursor cells or differentiated neuronal cells. However, the process of transmitter phenotype determination in the CNS is only beginning to be understood, with more uncharacterized substances, with considerable potency in this respect being reported and in need of isolation and further study. These studies will bring great advances in our existing knowledge of brain development and have potential value for the development of new treatments for neurodegenerative diseases. PMID- 9330423 TI - Habituation: events in the history of its characterization and linkage to synaptic depression. A new proposed kinetic criterion for its identification. AB - Reports on habituation from the last part of the nineteenth and the first part of this century are reviewed. Publications are selected according to their significance for the understanding of the properties that characterize habituation. Reports that have contributed to the establishment of a causative link between habituation and synaptic depression are also reviewed. A kinetic analysis of experimental cases of habituation is made for different reflexes in a number of species. The results of the analysis indicate the existence of two distinct subcategories of habituation: a slow and a fast version. Each category has a remarkably narrow range of kinetic variability, regardless of species and type of reflex. In one particular reflex, the tentacle withdrawal of Helix pomatia, both versions of habituation are displayed in response to different stimulation paradigms. On the basis of the kinetic analysis, a new criterion for identification of habituation is suggested. PMID- 9330424 TI - On the significance of temporally structured activity in the dorsal lateral geniculate nucleus (LGN). AB - Higher organisms perceive information about external or internal physical or chemical stimuli with specialized sensors that encode characteristics of that stimulus by a train of action potentials. Usually, the location and modality of the stimulus is represented by the location and specificity of the receptor and the intensity of the stimulus and its temporal modulation is thought to be encoded by the instantaneous firing rate. Recent studies have shown that, primarily in cortical structures, special features of a stimulus also are represented in the temporal pattern of spike activity. Typical attributes of this time structure are oscillatory patterns of activity and synchronous discharges in spatially distributed neurons that respond to inputs evoked by a coherent object. The origin and functional significance of this kind of activity is less clear. Cortical, subcortical and even very peripheral sources seem to be involved. Most of the relevant studies were devoted to the mammalian visual system and cortical findings on temporally structured activity were reviewed recently (Eckhorn, 1994, Progr. Brain Res., Vol. 102, pp. 405-426; Singer and Gray, 1995, Annu. Rev. Neurosci., Vol. 18, pp. 555-586). Therefore, this article is designed to give an overview, especially of those studies concerned with the temporal structure of visual activity in subcortical centers of the primary visual pathway, which are the retina and the dorsal lateral geniculate nucleus (LGN). We discuss the mechanisms that possibly contribute to the generation and modulation of the subcortical activity time structure and we try to relate to each other the subcortical and cortical patterns of sensory activity. PMID- 9330425 TI - Smooth non-parametric receiver operating characteristic (ROC) curves for continuous diagnostic tests. AB - Receiver operating characteristic (ROC) curves have seen increasing use to assess the accuracy of diagnostic tests that yield continuous test results. We can calculate a fully non-parametric ROC curve from the empirical false positive rate (FPR) and true positive rate (TPR) at each possible decision threshold, but it may be quite jagged. We can obtain a smooth ROC curve by directly fitting a parameteric model, for example, binormal or bilogistic, to the actual test results, but substantial lack-of-fit may result if the distributional assumptions are not valid. A recently proposed alternative algorithm 'LABROC4' is insensitive to such departures, but is not fully flexible in that the form of the resulting ROC curve is restricted to be binormal. We propose a smooth non-parametric ROC curve derived from kernel density estimates of the two test result distributions. We obtain pointwise standard errors for the estimated TPR at each FPR and use them to construct pointwise confidence intervals for the ROC curve, using a logit transformation. We also obtain standard errors for the estimated TPR and FPR at given thresholds and use them to construct confidence rectangles in (FPR, TPR) space that correspond to a specific threshold. We adapt existing methods for the unsmoothed non-parametric ROC curve to obtain the area under the ROC curve and its standard error; we also give partial areas and corresponding standard errors. We have created a FORTRAN algorithm 'ROC-&-ROL', available upon request. We compare ROC curves and areas obtained by applying our methods and its competitors to two data sets, one of which is fit well by parametric methods and LABROC4, the other of which is not. PMID- 9330426 TI - Effects of dependent errors in the assessment of diagnostic test performance. AB - Latent class models can be used to assess diagnostic test performance when there is no perfectly accurate gold standard test available for comparison. These models usually assume independent errors between the tests, conditional on the true disease state of the subject. Maximum likelihood estimates of the prevalence of the disease and the error rates of diagnostic tests are then obtained. This paper examines the impact of error dependencies between binary diagnostic tests on the parameter estimates obtained from the latent class models. The independence model often gives parameter estimates having relatively small bias, but in some situations (for example, when disease prevalence is low and the tests have low specificity, such as in population screening) the bias may be more serious. PMID- 9330427 TI - Estimating correlations from epidemiological data in the presence of measurement error. AB - Analysis of a major multi-site epidemiologic study of heart disease has required estimation of the pairwise correlation of several measurements across subpopulations. Because the measurements from each subpopulation were subject to sampling variability, the Pearson product moment estimator of these correlations produces biased estimates. This paper proposes a model that takes into account within and between sub-population variation, provides algorithms for obtaining maximum likelihood estimates of these correlations and discusses several approaches for obtaining interval estimates. PMID- 9330428 TI - A Markov model for HIV disease progression including the effect of HIV diagnosis and treatment: application to AIDS prediction in England and Wales. AB - Back-calculation is a widely used method to estimate HIV incidence rates, and is commonly based on times of AIDS diagnosis. Following up earlier work, we extend this method to also incorporate knowledge of times of HIV diagnosis (first positive test). This is achieved through the use of a Markov model which describes the progress of an HIV infected person through various stages, and which allows causal connections between events to be explicitly modelled. Estimation is based on maximum likelihood, the likelihood being calculated within a discretized version of the Markov model. The effect of sampling uncertainty and model uncertainty (sensitivity) is evaluated simultaneously by means of a combined bootstrap and simulation procedure. At each replication we resample both the data and the model (from a set of possible models described by randomizing one or more parameters). For instance, uncertain knowledge about the incubation distribution affects the estimates of some parameters, but not others. The Markov approach is applied to the prediction of AIDS incidence for homosexuals in England and Wales up to the year 2000. PMID- 9330429 TI - Classification of the effectiveness of combination treatments. AB - According to FDA regulations, a combination drug is not efficacious unless each component contributes to the claimed effects. For a univariate endpoint, this implies that the combination at specific doses must be superior to each of its components at the same doses. More demanding is the property of synergy, in which the effect of the combination must be superior to the effect expected based on those of its components. If it is equal to those effects, it is additive, and if it is inferior, it is antagonistic. We give regions in the combination dose plane where these concepts are well defined. If the effect of the combination is greater than the greatest effect achievable by any of its components it is therapeutically synergistic. A combination can be antagonistic, yet its components can still contribute to the claimed effects. If it is additive, synergistic or therapeutically synergistic, its components must contribute to the claimed effects. We relate these concepts and provide designs and sequential procedures for determining whether a combination is therapeutically synergistic, synergistic, additive, antagonistic and contributing or antagonistic and non contributing. PMID- 9330430 TI - A practical approach for the assessment of bioequivalence under selected higher order cross-over designs. AB - The two-period cross-over design with two sequences of drug administration is a standard experimental design when bioequivalence of one test formulation is to be assessed in comparison with a reference formulation. Previously, an approach based on Fieller's confidence interval has been presented for the assessment of average bioequivalence under this particular design. However, the two-sequence two-period cross-over design is not very useful in the presence of unequal carry over effects. Besides, this basic design does not provide independent estimators for the intra-subject variabilities. To overcome these limitations, it might be of interest to consider a higher-order cross-over design in which the number of periods and/or the number of sequences is greater than the number of formulations to be compared. Because of this, the present communication will concentrate on the generalization of Fieller's confidence interval concept for a particular group of higher-order cross-over designs. In addition to this, since the evaluation of simple average bioequivalence does not guarantee that the two products can be used interchangeably, the assessment of population and individual bioequivalence is addressed through the application of a comprehensible three step decision rule. An example study with a two-sequence four-period design is also analysed to illustrate the use of the proposed methods. PMID- 9330431 TI - A sequential design for psychophysical experiments: an application to estimating timing of sensory events. AB - An experimental subject sequentially receives different levels of a stimulus, and data are recorded on response or non-response to the stimulus. To ensure that the subject cannot predict the next stimulus level based on previous stimulus levels, a randomized design, based on a generalized Polya urn model, is used to allocate the stimulus levels. The goal of the experiment is to elicit information efficiently about the relationship between stimulus level and response (either for an individual subject or a group of independent subjects), by estimating quantiles of the stimulus-response curve. Our design allocates stimulus levels unimodally and symmetrically around the unknown median of the stimulus-response curve. We discuss estimation under a broad family of distributions and also fully discuss design issues and options. This design was used for an experiment in neurophysiology in humans to estimate the timing of onset of kinesthetic stimuli. Such psychophysical studies can increase our understanding of normal and pathological function. We present data from that experiment. PMID- 9330432 TI - Evaluation of a new screening assay ProC Global for identification of defects in the protein C/protein S anticoagulant pathway. AB - In the present study a new assay, ProC Global, globally estimating the activity of the main plasma components of anticoagulant protein C/protein S pathway, was evaluated with respect to test characteristics and its sensitivity in the detection of deficiency states of protein C and protein S and of increased aPCR. In the ProC Global assay procedure protein C is activated in patient's plasma by an activator reagent (venom from agkistrodon contortrix). The extent of the prolongation of a sample's aPTT, caused by the activation of protein C, is taken as a measure for its anticoagulant capacity. Ninety-eight patients with one of the above mentioned defects were investigated. Decreased plasma protein C activity and increased aPCR were detected with a sensitivity of 1.0, while only 11 of 14 patients with decreased levels of free protein S antigen showed abnormal results in the ProC Global assay (sensitivity = 0.79). The test can be used in heparinized samples up to 1.0 anti Xa U/ml heparin (UFH and LMWH). When samples from patients on oral anticoagulant treatment are prediluted with factor V deficient plasma the test is sensitive for increased aPCR. PMID- 9330433 TI - Bradykinin generation in RC-MAP during storage at 4 degrees C and leukocyte removal filtration. AB - We investigated contact system activation in red cell concentrate in mannitol adenine-phosphate (MAP) solution (RC-MAP) during storage and leukocyte removal filtration. The contact system activation was assessed in terms of bradykinin (BK) generation. The BK level in the RC-MAP transiently but significantly (p < 0.05) increased at 7 to 14 days of storage. Moreover, the BK level in 21-day stored RC-MAP was approximately 10 times higher than that in other blood components. The BK level in mannitol-containing MAP solutions, but not that in mannitol-free MAP solution, saline or whole blood, increased during storage. The plasma angiotensin-converting enzyme (ACE) activity decreased significantly only in mannitol-containing MAP solutions. These findings suggest that mannitol plays an essential role in the increase in BK level in RC-MAP. Furthermore, the BK level in RC-MAP increased 10 minutes after the start of filtration through a negatively charged filter, either a BPF-4 or an RN-40, and reached a maximum of 6,000 pg/ml. Hypotensive reactions are rare in RC-MAP transfusion in comparison to their incidence in platelet concentrate (PC) transfusion in spite of the higher BK level in RC-MAP than in PC. Therefore, BK generation in various blood components is not likely to be the main cause of hypotensive reactions. However, the level of BK in RC-MAP is sufficiently high to cause site pain. Further investigation on the clinical significance of a high BK level in blood components is necessary. PMID- 9330434 TI - Increased prevalence of a polymorphism in the gene coding for human prothrombin in patients with coronary heart disease. AB - A number of genetic risk factors for the development of coronary heart disease has been identified in the past. Some of these represent polymorphisms in genes of proteins which are associated with the process of blood clotting. We investigated the distribution of a recently described G/A polymorphism in the 3'untranslated region of the human prothrombin gene (nt 20210) in 98 patients (19 female age: 53 + 12 SD years and 79 male, age: 49 + 8.5 SD years) with coronary heart disease and in 102 healthy newborns by enzymatic amplification of the genomic DNA carrying the polymorphic site and by subsequent restriction digest. The diagnosis of coronary heart disease was established by coronary angiography in all patients. The frequency of the A allele in the healthy newborns was 0.98% (0.2%-3.5%; CI 0.95) with the G/A genotype occurring in 1.96% (0.24%-6.9%; CI 0.95). In the group of patients with coronary heart disease the G/A genotype was found in 5.1% (1.7%-11.4%; CI 0.95). 94.9% of the patients studied showed a G/G genotype. The A/A genotype was neither detected in the newborns nor in the patients with coronary heart disease. This preliminary study strongly suggest that the presence of the G/A polymorphism in the 3'untranslated region of the gene coding for human prothrombin is associated with the occurrence of coronary heart disease. PMID- 9330435 TI - Lipoprotein(a) concentration and molecular weight of apolipoprotein(a) in patients with cerebrovascular disease and diabetes mellitus. AB - Plasma lipoprotein(a) [Lp(a)] concentrations are genetically determined, and hyper-Lp(a)-emia is an independent risk factor for atherosclerosis and thrombosis. To study the implications of Lp(a) in cerebrovascular disease (CVD) and diabetes mellitus (DM), we examined plasma Lp(a) levels and molecular weights of apolipoprotein(a) [apo(a)] in 118 patients with CVD, and 125 cases with DM. Although mean Lp(a) concentrations were higher in those cases with atherothrombotic brain infarction than in those with brain hemorrhage and lacunar infarction, the difference was not statistically significant. Lp(a) levels were significantly higher in the DM cases treated with insulin and in those treated with oral hypoglycemic agents than in those on diet therapy alone, suggesting that insulin and oral agents modulate apo(a) expression. Lp(a) concentrations correlated significantly with the low-molecular-weight isoforms of apo(a) in all CVD and DM groups. PMID- 9330436 TI - Persistence of hemostatic alterations in patients affected by Crohn's disease after bowel surgery. AB - In Crohn's disease (CD) a condition of hypercoagulability with increased risk for thrombotic events has been reported. In this study we have investigated hemostatic parameters in thirty-one patients affected by CD before, 3 and 12 months after bowel operation, and in thirty healthy controls. Before surgery platelet number (PLT), fibrinogen (Fbg), prothrombin fragment F1 + 2 (F1 + 2), PAI and whole blood-spontaneous platelet aggregation (WB-SPA) were significantly higher (p at least < 0.0005) in patients than in controls, while factor XIII (F XIII) was significantly lower (p at least < 0.005). Three and twelve months after surgery PLT, FBG and WB-SPA significantly decreased in comparison to pre-surgery values (respectively p at least < 0.05 and p < 0.01), but PLT and Fbg were still significantly higher than in controls at 3 and 12 months (p < 0.01). At three and 12 months after operation F XIII was significantly higher in comparison with pre surgery values (p at least < 0.05). The presence of antiphospholipid antibodies (aPL) was not different between CD patients and controls before surgery, whereas it significantly increased 12 months after surgery (p < 0.05). Our results suggest that in CD hemostatic changes are only in part influenced by local flogistic processes and that an inflammatory systemic condition may provoke both the bowel and extraintestinal manifestations of CD. PMID- 9330437 TI - Involvement of cyclic GMP in the mode of action of a new antithrombotic agent PCA 4230; inhibition of the platelet cyclic GMP dependent phosphodiesterase. AB - The effect of PCA-4230, a new dihydropyridine derivative with a potent antithrombotic activity, on cyclic nucleotide phosphodiesterase in platelets was studied. PCA-4230 inhibited (54%) cyclic GMP hydrolytic activity of a platelet cytosolic fraction, whereas it did not affect that of cyclic AMP. Results suggested that PCA-4230 inhibited a cyclic GMP-dependent phosphodiesterase, known as cGB PDE or type V, on a purified enzyme from rabbit platelets by a non competitive-uncompetitive type inhibition. In addition, PCA-4230 potentiated the increase in both cyclic GMP and cyclic AMP levels evoked by sodium nitroprusside. Furthermore, PCA-4230 and forskolin caused a synergistic effect in cyclic AMP, and also potentiated the phosphorylation of 50 kDa and 22 kDa proteins, reported as substrates of cyclic GMP- and cyclic AMP-dependent protein kinases that are related to the inhibition of platelet functions. Finally, PCA-4230 also potentiated the forskolin- and sodium nitroprusside-inhibited serotonin release evoked by thrombin, probably related to the increased cyclic nucleotide level. PMID- 9330438 TI - The effect of physical conditioning suggests adaptation in procoagulant and fibrinolytic potential. AB - Acute exercise evokes a transient increase in procoagulant activity. We evaluated the effect of physical conditioning on the activation of the coagulation and fibrinolytic systems. Two groups of subjects of different aerobic endurance levels (athletes and controls with maximal oxygen uptake (VO2max) 68.4 and 52.6 ml kg-1 min-1, respectively), were tested at rest and after standardized exercise at 80% of their individual VO2max. There was a significant increase in prothrombinfragment 1 + 2 (F1 + 2) level among controls in response to standardized exercise (p < 0.05), whereas no significant difference in the level of F1 + 2 between athletes and controls at rest or in response to exercise was demonstrated. Tissue plasminogen activator (tPA) antigen level at rest was significantly lower in athletes compared to controls (p < 0.03). A significant increase was found in the tPA level after standardized exercise in both groups (p < 0.02), which was lower in athletes compared to controls (p < 0.05). There were no significant differences between athletes and controls in plasminogen activator inhibitor-1 (PAI-1) and thrombin antithrombin complex (TAT) levels at rest. Athletes had a significantly lower PAI-1 level than controls after exercise (p < 0.05). In conclusion, the present study suggests an increased activation of the coagulation system in response to exercise in controls only. It also suggests adaptive changes in fibrinolytic potential induced by physical conditioning, as demonstrated by the lower level of tPA at rest and the lower levels of tPA and PAI-1 after exercise in athletes compared to controls. PMID- 9330439 TI - Enhancement of plasminogen binding and fibrinolysis by chloropeptin I. AB - Plasminogen is a zymogen of the fibrinolytic serine protease, plasmin. Plasminogen binds, through its lysine binding sites in the kringle domain, to blood and vascular cells or fibrin, where it is efficiently activated and exerts fibrinolytic activity (1,2). We have recently found that complestatin, a peptide like metabolite of streptomyces (3,4), enhances plasminogen binding to U937 cells and fibrin, thus potentiating fibrinolysis (5). In the present study, complestatin was found to be converted by an acid treatment to a more active isomer in enhancing plasminogen binding to U937 cells. This isomer was identified to be chloropeptin I, which was recently isolated from a culture of Streptomyces sp. by Matsuzaki et al. as an inhibitor of gp 120-CD4 binding (6). The present paper deals with the stimulation of fibrinolysis by chloropeptin I. PMID- 9330440 TI - The effect of delayed analysis or freeze-thawing on the measurement of natural anticoagulants, resistance to activated protein C and markers of activation of the haemostatic system. AB - We provide a centralised thrombophilia screening service with sample transfer by courier or first class mail, a practice common to many centres. Sample quality is of prime importance, thus we have assessed the effect of delayed sample handling upon the haemostatic variables within our thrombophilia profile. This comprises screening for familial natural anticoagulant deficiency and APC resistance (1-4), and acquired lupus anticoagulant (5,6). The effect upon hypercoagulable markers were also assessed to facilitate evaluation of these in prospective clinical studies (7). PMID- 9330441 TI - Contrast ultrasonography for 2-D opacification of heart cavities, peripheral vessels, kidney and muscle. AB - Contrast ultrasonography of peripheral vessels and peripheral organs has been only sparsely used to evaluate peripheral tissue blood flow. The purpose of the study was to characterize intraluminal opacification of renal and femoral arteries and veins, of skeletal muscle and renal parenchyma after intraarterial (IA) injection of BY963, a newly developed ultrasound contrast agent being evaluated in Phase II and III trials, and to compare it with opacification of heart cavities after intravenous injection (IV) in dogs. A further purpose was to quantitate possible opacification losses during the first transcapillary passage of BY963 through pulmonary and peripheral microcirculation. BY963 was administered at the dose of 5 mL/animal/vascular territory (0.2 mL/kg). The peak intensity (intensity units = IU) and the area-under-the-curve (AUC, IU x heart cycles) were estimated from regions-of-interest placed in the right ventricle (RV), left ventricle (LV), main renal artery and vein, kidney, femoral artery and vein and adductor muscle. Following single IV injection, the average peak intensity and AUC values were 33 +/- 3 (mean +/- SE) and 674 +/- 109 for the RV, and 27 +/- 2 and 870 +/- 74 for the LV (p < 0.05), respectively. Following single IA injection in the descending aorta, the average peak intensities and AUC values were 35 +/- 2 and 613 +/- 139 in the renal artery and 26 +/- 4 (p < 0.05) and 639 +/- 151 in the renal vein (nonsignificant), respectively. For the femoral vessels, the average peak intensities and AUC values were 30 +/- 3 and 469 +/- 63 in the femoral artery, and 21 +/- 2 (p < 0.05) and 517 +/- 44 in the femoral vein (nonsignificant), respectively. The values for the output-to-input intensity ratios for peak intensity and AUC were 0.82 +/- 0.06 and 1.36 +/- 0.12 for the LV/RV ratio, 0.73 +/- 0.08 and 1.02 +/- 0.05 for the renal vein/renal artery ratio, and 0.71 +/- 0.09 and 1.16 +/- 0.13 for the femoral vein/femoral artery ratio, respectively (nonsignificant). In conclusion, these results demonstrate the high opacification potency of BY963 in the LV, renal and femoral veins, being of the same order of magnitude as that in the RV, renal and femoral arteries, respectively. Finally, the loss of opacification properties of BY963 during the first transcapillary (pulmonary or peripheral-capillary) passage is minimal. PMID- 9330442 TI - Stiffness and diameter of the common carotid artery and abdominal aorta in women. AB - Vascular disease is differentiated throughout the vascular regions with aorta more prone to dilatation and with peripheral arteries more prone to occlusive disease. In this study, we investigated the diameter and distensibility in the common carotid artery (CCA) and abdominal aorta (AO) in healthy females of varying ages to assess potential differences in the aging process. The diameter and pulsatile diameter change of the CCA and AO in 66 healthy Caucasian females aged 8 to 79 y were examined using an ultrasound phase-locked echo-tracking technique. Blood pressure was obtained by the auscultatory method. Arterial stiffness (beta) was calculated. The diameter of both CCA and AO increases, not only in childhood, but also in adult women. The dilatation in adults (25 to 75 y) seems to be more pronounced in the AO (23%) than in the CCA (11%). Stiffness (beta) is higher in the CCA than AO before 20 y of age (p < 0.05). Stiffness increases with aging in both arterial regions, but the increase is initially somewhat higher in the AO, which means that no differences between the CCA and AO are seen from 25 y onward. In conclusion, this study demonstrates regional differences in diameter change and stiffness in the CCA and AO, and implies that the AO is more prone to age-related changes of the arterial wall than the CCA. These differences, however, are not as marked as previously reported in males. PMID- 9330443 TI - Ultrasonic characterization of tissues via backscatter frequency dependence. AB - Phantom and patient studies were performed to assess the potential of backscatter frequency dependence as a useful parameter for tissue characterization. A commercial phased-array ultrasonic scanner was adapted to allow digitization of the intermediate-frequency ultrasonic data. Studies of agar phantoms containing polystyrene microspheres with 3.5 and 5 MHz transducers indicated the ability for robust differentiation of phantoms having different scatterer size and frequency dependence, based on calculated differences in mean frequencies of backscattered spectra. Using a 3.5-MHz probe, significantly lower mean frequency of ultrasound backscattered from cirrhotic, compared to normal, liver tissue was noted. Studies of benign and malignant liver tumors (hemangiomas and metastases, respectively) indicated differences in frequency content of these tumors, compared to the adjacent normal liver, that were statistically significant for 3.5-MHz data. PMID- 9330445 TI - Automated detection of local artery wall thickness based on M-line signal processing. AB - The Young's modulus of an arterial segment, a measure of the elastic properties of the arterial wall, requires the simultaneous and local assessment of pulse pressure, wall thickness, diameter, and distensibility (relative increase in cross-sectional area per change in blood pressure). The diameter and relative increase in cross-sectional area can be obtained with a wall track system, processing the radiofrequency (r.f.) ultrasound signals received along a single line of observation (M-line processing). It will be demonstrated that it is feasible to combine, in a single measurement, the assessment of wall thickness and the (relative change in) diameter involving a minimum of user interaction. Phantom tests show a standard error of the estimate for intima-media thickness measurements of less than 20 microns; in vivo registrations exhibit a variation on the order of 45 microns. It is concluded that processing of the radiofrequency ultrasound signal, acquired along an M-line, provides an accurate and time efficient alternative for videoprocessing of 2-dimensional B-mode ultrasound images to estimate artery wall thickness. PMID- 9330444 TI - A high-frequency pulsed-wave Doppler ultrasound system for the detection and imaging of blood flow in the microcirculation. AB - Previous work with a 40-MHz continuous-wave Doppler ultrasound system has demonstrated the potential of high-frequency Doppler ultrasound (HFD), operating in the frequency range 20-200 MHz, to detect blood flow in the microcirculation. This paper describes a directional, pulsed-wave high-frequency Doppler ultrasound (PW HFD) system that was designed and constructed further to investigate this potential. The PW HFD system electronics have a dynamic range of > 80 dB, a noise floor of 250 nV, a directional isolation of 45 dB and operate over the frequency range 1-200 MHz. The system is tested using a focused PVDF transducer that is tuned for maximum sensitivity at 50 MHz, has a -6 dB lateral beamwidth of 70 microns and -6 dB depth-of-focus of 0.90 mm. This permits the practical use of sample volumes with dimensions 70 microns laterally by 90-900 microns axially. The PW HFD system can operate in duplex mode by either sharing the Doppler transducer or using a second PVDF transducer for imaging. Tests with string and capillary flow phantoms demonstrate that the PW HFD system is capable of detecting velocities on the order of the blood velocities found in the capillaries (0.5 mm/s) and arterioles (5 mm/s) with suitable velocity (30-300 microns/s) and temporal (15-100 ms) resolutions. In vivo measurements demonstrate that PW HFD can detect and measure blood velocities of less than 5 mm/s in arterioles and venules with diameters as small as 20 microns and 35 microns, respectively, using a sample volume of only 70 microns laterally by 150 microns axially. Preliminary experiments with high-frequency colour Doppler (HFCD) and high-frequency power Doppler (HFPD) imaging are also presented. PMID- 9330446 TI - Evaluation of cerebral arterial flow with transcranial Doppler ultrasound: theoretical development and phantom studies. AB - Blood flow information available from transcranial Doppler ultrasound is usually derived from velocity alone because no knowledge of vessel caliber is available. In cases such as vasospasm, where vessel size changes, the inference of flow from velocity becomes questionable. A computational technique was used to calculate a flow index and 2 vessel area indices based on the first and zero moments of the Doppler power spectrum. These indices were tested in a steady and pulsatile flow phantom using 6 different diameter elastic tubes. Changes in the flow index showed good agreement with changes in timed volume flow for different flow rates. The vessel caliber indices correctly predicted changes in area when different diameter tubes were examined. These indices may prove useful in clinical settings where the constancy of flow or vessel diameter between studies are in question. PMID- 9330447 TI - A study of the spectral broadening of simulated Doppler signals using FFT and AR modelling. AB - Doppler ultrasound is used clinically to detect stenosis in the carotid artery. The presence of stenosis may be identified by disturbed flow patterns distal to the stenosis that cause spectral broadening in the spectrum of the Doppler signal around peak systole. This paper investigates the behaviour of the spectral broadening index (SBI) derived from wide-band spectra obtained using autoregressive modelling (AR), compared with the SBI based on the fast-Fourier transform (FFT) spectra. Simulated Doppler signals were created using white noise and shaped filters to analyse spectra typically found around the systolic peak and to assess the magnitude and variance of AR and FFT-SBI for a range of signal to-noise ratios. The results of the analysis show a strong correlation between the indices calculated using the FFT and AR algorithms. Despite the qualitative improvement of the AR spectra over the FFT, the estimation of SBI for short data frames is not significantly improved using AR. PMID- 9330448 TI - Ultrasonic measurement of breast tissue motion and the implications for velocity estimation. AB - A high-resolution study of breast tissue motion during cardiac systole and respiration is presented. An experimental system was designed to achieve a velocity resolution on the order of 1 mm/s with high spatial resolution. The peak velocity of tissue motion estimated during cardiac systole ranged from 0.2 mm/s to 5.6 mm/s among the subjects studied. It is shown that motion due to the cardiac cycle is less significant when the subject is positioned on the side rather than supine. The mean tissue velocity among subjects in the supine position is 2.88 mm/s and drops to 0.81 mm/s for the side position, with a corresponding spatial displacement of 0.095 mm, dropping to 0.027 mm. The velocity profiles indicate that 100 ms is required for the entire ribcage contraction-relaxation process to occur. Experiments using a prone biopsy table show the almost complete elimination of tissue motion due to cardiac systole, suggesting that the use of the table eliminates this motion, thus allowing for high-resolution blood velocity estimates. Features resulting from respiratory motion are also presented. We found this motion to be of a much longer time duration, and of a much higher magnitude, with velocities as high as 29 mm/s. The implications of the study on the high-resolution estimation of blood velocity and high-resolution breast imaging are discussed. PMID- 9330449 TI - Influences of ultrasonic machine settings, transducer frequency and placement of region of interest on the measurement of integrated backscatter and cyclic variation. AB - Integrated backscatter and its cyclic variation are potentially important parameters to discriminate normal from diseased myocardium. Cyclic variation of integrated backscatter is expected to be independent of machine settings. Backscatter images of swine hearts were taken using a two-dimensional backscatter system while acoustic power was varied at different time gain control (TGC) settings. Cyclic variation was measured in vivo with various acoustic power and TGC settings using different transducer frequencies. Three different regions were analyzed. For any given TGC setting, the relationship between acoustic power and integrated backscatter in vitro was linear only over a narrow range. In vivo, cyclic variation was present at all regions studied in both long- and short-axis views. However, lower acoustic power (< 15 dB) and TGC (< 20 dB), or excessive settings of acoustic power (> 35 dB) and TGC (> 50 dB), produced minimal cyclic variation. Appropriate acoustic power (20-35 dB) and TGC (30-50 dB) produced larger and more consistent cyclic, variation at the posterior region of the left ventricle. These data indicate that each region has specific, appropriate machine settings to maximize the magnitude of cyclic variation. PMID- 9330450 TI - Influence of tissue preservation methods on arterial geometry and echogenicity. AB - Thoracic porcine aortas from 5 pigs were investigated with 7.5-MHz ultrasound in vitro at low and high transmural pressure before and after the following tissue preservation methods were applied: 1. Storage in frozen condition (-12 degrees C) for 24 h followed by thawing; 2. fixation in formalin at zero transmural pressure for 24 h; and 3. fixation with formalin for 24 h while applying 74 mmHg of transmural pressure from within the lumen and a tensile force to longitudinally stretch the artery. Fixation in formalin at zero transmural pressure resulted in swelling of the arterial wall (25 +/- 40%, p < 0.02, at low transmural measurement pressure) and in decreased echogenicity (-23 +/- 38%, p < 0.01) of the arterial vessel wall. No changes in this respect were found after storage in a frozen condition nor after fixation in formalin at high transmural pressure which, therefore, are more appropriate procedures for fixation of arteries prior to in vitro ultrasound examination if geometry is important. PMID- 9330451 TI - A numerical study of the effect of drive level on the intensity loss from an ultrasonic beam. AB - Knowledge of the spatial distribution of intensity loss from an ultrasonic beam is vital to bioeffect predictions such as heating and streaming. A method for calculating the distribution of intensity loss over a finite-amplitude ultrasonic beam is described. The technique demonstrates that the location of peak intensity loss varies considerably with drive level for a plane circular source, as does the overall distribution of intensity loss across the beam. The effects are shown to be less pronounced but still significant for a focused source. PMID- 9330452 TI - Stress-wave-induced membrane permeation of red blood cells is facilitated by aquaporins. AB - Stress waves generated by lasers and extracorporeal lithotripters have been shown to transiently increase the permeability of the plasma membrane, without affecting cell viability. Molecules present in the medium can diffuse into the cytoplasm under the concentration gradient. Molecular uptake under stress waves correlates with the presence of functioning aquaporins in the plasma membrane. PMID- 9330453 TI - Mechanism of ultrasound enhanced porphyrin cytotoxicity. Part I: A search for free radical effects. AB - Intense ultrasound beams may have the potential to treat malignant tumours when combined with sensitizers, often called sonodynamic agents. Some of these agents, e.g., the porphyrins, are currently used for photodynamic therapy. However, the experimental evidence for ultrasound activation of sensitizers is inconsistent. This paper attempts to discover whether they yield of free radicals such as .OH and .H, which are produced by transient cavitation, could explain the killing of Chinese hamster ovary (CHO) cells in vitro with and without sonodynamic agents. CHO cells were irradiated with ultrasound beams in phosphate-buffered saline or in growth medium, and the immediate cell lysis and loss of cell colony forming ability were measured. Under our specific conditions, in which the standing wave patterns were minimized, a general correlation was observed between the transient cavitation, free radical production, and cytotoxicity. However, the yield of free radicals was much too small to explain the cell killing observed. We conclude that cytotoxicity is not linked to attack from free radicals formed outside the cells. In our experiments, immediate cell lysis is closely linked to the transient cavitation, which is known to produce shear forces that disrupt cellular membranes. We hypothesize that the loss of cell colony forming ability is also linked to damage of cellular membranes. PMID- 9330454 TI - In vitro study of the mechanical effects of shock-wave lithotripsy. AB - Impulsive stress in repeated shock waves administered during extracorporeal shock wave lithotripsy (ESWL) causes injury to kidney tissue. In a study of the mechanical input of ESWL, the effects of focused shock waves on thin planar polymeric membranes immersed in a variety of tissue-mimicking fluids have been examined. A direct mechanism of failure by shock compression and an indirect mechanism by bubble collapse have been observed. Thin membranes are easily damaged by bubble collapse. After propagating through cavitation-free acoustically heterogeneous media (liquids mixed with hollow glass spheres, and tissue) shock waves cause membranes to fail in fatigue by a shearing mechanism. As is characteristic of dynamic fatigue, the failure stress increases with strain rate, determined by the amplitude and rise time of the attenuated shock wave. Shocks with large amplitude and short rise time (i.e., in uniform media) cause no damage. Thus the inhomogeneity of tissue is likely to contribute to injury in ESWL. A definition of dose is proposed which yields a criterion for damage based on measurable shock wave properties. PMID- 9330455 TI - Measurement of shear-wave velocity by ultrasound critical-angle reflectometry (UCR). AB - There exists a growing body of research that relates the measurement of pressure wave velocity in bone to different physiological conditions and treatment modalities. The shear-wave velocity has been less studied, although it is necessary for a more complete understanding of the mechanical properties of bone. Ultrasound critical-angle reflectometry (UCR) is a noninvasive and nondestructive technique previously used to measure pressure-wave velocities both in vitro and in vivo. This note describes its application to the measurement of shear-wave velocity in bone, whether directly accessible or covered by soft tissue. PMID- 9330456 TI - Infectious disease on the run. A report on the Children's Vaccine Initiative (CVI): consultative group meeting, Dakar, Senegal, 9-10 December 1996. PMID- 9330457 TI - CVI awards celebrate the Year of the Vaccine and recent contributions in immunization and vaccine development. PMID- 9330458 TI - Vaccine development against dengue and Japanese encephalitis: report of a World Health Organization meeting. PMID- 9330459 TI - Oral immunization with recombinant vaccina expressing cell-surface-anchored beta hCG induces anti-hCG antibodies and T-cell proliferative response in rats. AB - A vaccinia recombinant, VSS2, expressing cell-surface-anchored beta-subunit of human chorionic gonadotropin (beta hCG) has earlier been found to induce high titered anti-hCG antibodies in rats immunized by tail scarification. The immunogenicity of this recombinant was compared in rats which received the virus through different routes of inoculation: intradermal, intragastric, intrajejunal or by tail scarification. The recombinant virus induced high titers of anti-hCG antibodies in all instances although the titers were about one log lower when the recombinant virus was delivered orally. The recombinant was found to induce T cell proliferative response in rats of all immunization groups. PMID- 9330460 TI - Influenza vaccination in 22 developed countries: an update to 1995. AB - This study expands and updates through 1995 our earlier report on influenza vaccine use in 18 developed countries. Five of the six countries with high levels of vaccine use in 1992 (> or = 130 doses/1000 population) showed little change or slight declines over the subsequent 3 years. The exception was the United States, where a new federal program for vaccination reimbursement for the elderly helped to increase vaccine distribution from 144 to 239 doses/1000 population. The six countries with medium levels of vaccine use in 1992 (76-96 doses/1000 population) increased to > or = 100 doses/1000 population by 1995. Among the six low-use countries in 1992 (< or = 65 doses/1000 population), only Finland showed substantial improvement (96 doses/1000 population) in 1995. Four new countries were added to the study. In Germany, vaccine use increased to 80 doses/1000 population in 1995, but in Ireland it remained at a low level (48 doses/1000 population). In Korea, vaccine use increased from 17 to 95 doses/ 1000 population during the period 1987-1995. In Japan, very high levels of vaccine use (approximately 280 doses/1000 population) in the early 1980s were associated with vaccination programs for school children. However, vaccine use fell precipitously when these programs were discontinued, and only 2 and 8 doses/1000 population were used in 1994 and 1995, respectively. In all 22 countries, higher levels of vaccine use were associated with vaccination reimbursement programs under national or social health insurance and were not correlated with different levels of economic development. Excluding Japan, in 1995 there was still a greater than fourfold difference between the highest and lowest levels of vaccine use among the other 21 countries in the study. Given its well established clinical effectiveness and cost-effectiveness, none of these countries has yet achieved the full benefits of its programs for influenza vaccination. PMID- 9330461 TI - Inactivated bovine herpesvirus 1 marker vaccines are more efficacious in reducing virus excretion after reactivation than a live marker vaccine. AB - A comparative study was carried out to evaluate the efficacy of three bovine herpesvirus 1 (BHV1) marker vaccines to reduce the reexcretion of virus after reactivation of latent BHV1. A live gE-negative vaccine, an inactivated gE negative vaccine and an experimental gD-subunit vaccine were tested in three identical experiments in which cattle, latently infected with BHV1, were vaccinated twice before they were treated with high doses of dexamethasone. Virus excretion after dexamethasone treatment was compared with that in BHV1-infected, unvaccinated cattle which served as controls. All cattle, controls and vaccinees, excreted virus. However, the inactivated vaccines reduced virus excretion more efficiently than did the live vaccine. PMID- 9330462 TI - Antibody response to influenza immunization in patients after heart transplantation. AB - The aim of this study was to evaluate post-heart transplantation (Htx) response to two-dose and three-dose influenza vaccine. Hemagglutination inhibition antibodies were monitored in HTx recipients immunized twice (n = 25) or three times (n = 17), and non-HTx controls (n = 8) once, with inactivated influenza vaccine. Post-first dose protective antibody titers (> or = 1:40) were demonstrated in 9/25 (36%) for A/Singapore/ (H1N1), 5/25 (20%) for A/Shanghai/(H3N2) and 2/25 (8%) for B/Yamagata compared with 4/8 (50%), 6/8 (75%) and 2/8 (25%), respectively, for controls. Post-second dose protective titers remained low, increasing following the third dose to 71%, 65% and 29%, respectively. The abnormally low antibody responses of HTx recipients to one-dose and two-dose influenza vaccine can be overcome by a third dose. PMID- 9330463 TI - Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia). AB - Naja kaouthia (NK) causes the highest fatality due to snake venom poisoning in Thailand. The specific antivenom produced is of low potency and in short supply. The aim of this study was to improve the antivenom potency. Bentonite and complete Freund's adjuvants (CFA) and various immunogens were compared. Six groups of three to five horses were immunized as follows: Group 1, NK venom adsorbed on bentonite; Group 2, NK venom in CFA; Group 3, NK venom in CFA in multi-emulsion formulation; Group 4, NK venom in 25% CFA; Group 5, NK neurotoxin 3 (NK3) conjugated with tetanus toxoid (NK3-TT) in CFA; Group 6, NK3 conjugated with diphtheria toxoid (NK3-DT) in CFA. Horses in Group 2-6 produced antivenom of very high neutralizing activity, up to four times higher than that of horses of Group 1. CFA (100 or 25%) or as a multi-emulsion formulation, induced comparable neutralizing antibody production with all three immunogens. All horses showed normal weight gain during the course of immunization. Group 1 horses exhibited minimal local reactions while horses in the other five groups had mild and comparable local reactions at the injection sites. No significant differences in the reactions caused by CFA in different formulations or different immunogens were observed. The production of highly potent antivenom against N. kaouthia from these horses should help solve problems associated with the currently available antivenom. PMID- 9330464 TI - Erythrocytes enhance the immunogenicity of oral vaccination with gamma irradiated influenza virus: increasing the dose of irradiation results in a significant diminution of lung IgA response. AB - Previous studies have demonstrated that the ability of gamma-irradiated whole influenza virus to prime for specific anti-influenza antibody responses was dramatically enhanced when delivered in a complex with chicken red blood cell ghosts (cRBC). The purpose of this study was to investigate the effect of increasing the dose of gamma irradiation used to inactivate A/Queensland/6/72 virus on the ability of the virus-cRBC complex to prime for specific influenza responses. Spleen cell proliferation studies confirmed the enhancing effect of the cRBC carrier for oral vaccination with irradiated virus. Cells from mice vaccinated with 30 kGy-irradiated virus only did not respond to influenza stimulation in vitro, whereas cells from mice vaccinated with irradiated virus+cRBC showed significant increases in proliferation to antigen exposure. No significant antibody response or challenge virus clearance was observed in mice orally vaccinated with irradiated (13.1 or 30 kGy) virus alone, even when the dose was increased significantly. Oral vaccination with live virus (+/-cRBC) primed for significant influenza specific IgA responses in the lungs, in addition to IgG responses in the lungs and sera. The dose of irradiation used to inactivate the virus was found to be critical to the profile of antibody response when the virus was delivered in a complex with cRBC. Oral vaccination of Swiss mice with 13.1 or 30 kGy virus (+cRBC) primed for significant serum and lung IgG responses. Lung IgA responses for 13.1 kGy+cRBC vaccinated mice were detected, but 30 kGy+cRBC vaccinated Swiss and CBA/H mice had no significant lung IgA response. The abrogation of IgA response, however, did not lessen the clearance of live challenge virus in outbred mice, suggesting a primary role for IgG and/or CTL response in the control of influenza virus infection post oral vaccination. To ensure direct comparison of virus alone and virus+cRBC treatments, the concentration of virus complexed to the cRBC was determined. PMID- 9330466 TI - Human schistosomiasis: potential long-term consequences of vaccination programmes. AB - Fields trials of new schistosomiasis vaccines are anticipated within the next few years, but there remains great uncertainty regarding the optimal design of vaccination programmes. Mathematical models are used here to explore the potential long-term consequences of vaccination, assuming that the vaccines provide partial protection for a limited period. The analysis suggests that vaccines acting to reduce infection rates or egg output will have a similar impact on levels of infection, that this impact may be highly sensitive to the duration as well as the degree of protection, that it may take several decades for the full impact to become apparent, and that one consequence will be peak levels of infection occurring in older age classes. In terms of lowering levels of infection there may be advantages in delaying vaccination until children reach school age, especially if the vaccine gives short-lived protection, or to repeat vaccination. The short-term advantages can be greatly increased by combining the introduction of a vaccination programme with initial mass chemotherapy. Continuous combined vaccination and chemotherapy programmes may also be more effective than either intervention alone. More research is needed on the consequences of vaccinating previously vaccinated, infected, and infected and treated individuals and the importance of natural boosting of vaccine-induced immunity. PMID- 9330465 TI - Twenty years' experience of rubella vaccination in Sweden: 10 years of selective vaccination (of 12-year-old girls and of women postpartum) and 13 years of a general two-dose vaccination. AB - Two different strategies for the prevention of rubella-induced malformations have now been practised in Sweden, both reaching 90% or more of the target populations. The first was initiated in 1973-1974 and targeted schoolgirls, susceptible women after pregnancy and women at special risk. The second programme a two-dose measles, mumps, rubella vaccination (MMR) of both boys and girls at the ages of 18 months and 12 years-was introduced in 1982. The percentage of susceptible pregnant women was gradually reduced from 12% in 1975 to 2.8% in 1987 and to just below 2% in 1994. The majority of the non immune are unvaccinated, these being either Swedes born before 1963 or immigrants. Before 1974 on average 14 severely rubella damaged children were reported yearly. Between 1975 and 1985 only a mean of two cases per year were recorded. Since 1985 no child with the rubella syndrome has been registered. PMID- 9330467 TI - Immunization with a polyvalent OspA vaccine protects mice against Ixodes ricinus tick bites infected by Borrelia burgdorferi ss, Borrelia garinii and Borrelia afzelii. AB - Sequence variability of the outer surface protein (Osp) A among Borrelia burgdorferi sl species suggests that a monovalent OspA vaccine may not protect against the various Borrelia present in Eurasia. Here, we confirmed that a monovalent recombinant OspA (rOspA) vaccine does not protect mice against Ixodes ricinus mediated infection with B. burgdorferi ss, Borrelia garinii and Borrelia afzelii. However, when mice were vaccinated with a cocktail of various rOspA from these three species, they were protected, and all challenge ticks that fed on them were cleared of their spirochetes. These results showed that a multiple OspA antigens vaccine, compatible with human use, was very efficient at protecting mice against B. burgdorferi ss, B. garinii, and B. afzelii. PMID- 9330468 TI - Immunoglobulin E and G responses to pertussis toxin in children immunised with adsorbed and non-adsorbed whole cell pertussis vaccines. AB - The IgE and IgG responses to pertussis toxin were measured in blood samples from 70 children (age 1.5-2.9 years) after primary immunisation with either a non aluminium adsorbed, whole cell vaccine (n = 34) or an aluminium adsorbed whole cell vaccine (n = 36). Two years later, they received a booster immunisation with either the non-adsorbed (n = 24) or the aluminium adsorbed vaccine (n = 14). Neutralising antibodies to pertussis toxin were higher (P < 0.05) after the three priming doses of the adsorbed vaccine than of the non-adsorbed vaccine, although both groups showed > 90% seropositives after the third dose. IgE antibodies to PT (PT-IgE) were detected in samples from 11/52 children after completed primary immunisation and the levels were low (median < or = 0.1 PRU ml-1) in both groups. No significant differences between the groups were found. PT-IgE levels did not increase after the booster injection. Thus, the aluminium content of the whole cell vaccines influenced the IgG response but not the IgE responses to pertussis toxin. The high rates of PT-IgE responses noted after a booster dose of acellular or whole cell pertussis vaccine to children primed with acellular vaccine in previous studies can therefore be mainly ascribed to the nature of the priming vaccine rather than the aluminium adjuvant. PMID- 9330469 TI - Immunization against the murine malaria parasite Plasmodium yoelii using a recombinant protein with adjuvants developed for clinical use. AB - Mice vaccinated with a recombinant protein containing the two EGF-like modules of Plasmodium yoelii merozoite surface protein-1 in liposomes or combined with the formulations SBAS2.1 and SBAS2, were protected against a lethal malaria infection. The protection achieved with these adjuvants developed for clinical use was as good as or better than that achieved with Freund's adjuvant. A parasite-specific response was needed for protection. Analysis of the immunoglobulin sub-class response showed that MSP-1-specific IgG1, and to a lesser extent IgG2a and IgG2b, were induced, suggesting that these antibodies were important for protection. Mice passively immunized with serum or purified IgG from vaccinated mice had delayed onset of parasitemia and were able to control the infection. PMID- 9330470 TI - Confronting the degeneracy of convergent combinatorial immunogens, or 'mixotopes', with the specificity of recognition of the target sequences. AB - Immunization by convergent combinatorial peptide libraries, or 'mixotopes' represents an interesting approach for inducing broadly cross-reactive immune response to hypervariable pathogens. The authors have immunized rabbits with a series of eight HIV-1 V3-loop derived constructs of increasing complexity, and analysed the reactivity of the corresponding antisera towards a set of V3-related peptides. Results were surprisingly homogeneous. Mixotopes containing as many as several billion closely related combinatorial sequences were immunogenic, and able to induce V3-specific antibodies. These results suggest that serological cross-reactivity depends on the sequential similarity of the antigen with the parent immunogen. Such 'mixotopes' could represent a useful approach to vaccination against hypervariable pathogens. PMID- 9330471 TI - Identification of strain-specific nucleotide sequences in E1 and NS4 genes of rubella virus vaccine strains in Japan. AB - Strain-specific nucleotide sequences of E1 and NS4 genes in five strains of a live rubella virus vaccine manufactured in Japan were identified for comparison, using 2389 nucleotides (1443 nucleotides of the E1 gene, 41 of the 3' terminal region following the E1 gene and 905 of the NS4 gene). Sequences of the E1 gene in three strains (Matsuura, TCRB19 and To-336) were identified. Takahashi and Matsuba strains shared common sequences, but were discriminated by the sequence of the NS4 gene. These five strains showed a phylogenetic relationship with the places of their isolation. In a comparative study of three strains with their unattenuated progenitors, the nucleotides in these regions were almost conserved during the attenuation process. PMID- 9330472 TI - Induction of T1 (cytotoxic lymphocyte) and/or T2 (antibody) responses to a mucin 1 tumour antigen. AB - Effective vaccination-based control of intracellular pathogens or parasites and various tumours is dependent upon induction of cytotoxic lymphocytes and other mechanisms of cellular immunity. Such responses are usually described as being antagonistic to an antibody-based immune response. This paper elaborates on previous studies that have demonstrated that conjugation of a fusion protein (FP, incorporating copies of the variable number of tandem repeat sequence of human mucin-1 (MUC1)) to oxidized mannan results in a significant shift from a type-2 response towards a type-1 response. This response induces complete protection upon challenge of immunized mice with MUC1 expressing tumour cells. This report details experiments in which the balance between type-1 and type-2 anti-MUC1 responses is manipulated by altering the dose of mannan-FP (M-FP) delivered. It is also shown that type-1 and type-2 responses may be induced simultaneously by administration of both forms of the antigen (FP/M-FP). Further, when a type-2 response is induced after FP immunization, a type-1 response can also be established by subsequent immunization with M-FP without adversely affecting the initial response. The converse also applies when M-FP is used for the initial immunizations, followed by FP administration. Delivery of interleukin-1 beta as a cytokine adjuvant with M-FP immunizations also enhanced antibody responses to levels fourfold that induced by M-FP alone without adversely affecting the cytotoxic activity induced by M-FP immunization. Contrary to the type-1/type-2 paradigm, cellular and antibody responses to MUC1 were not antagonistic. These results have important implications for the development of vaccination strategies against pathogens for which both the cellular and humoral compartments of the immune response contribute to protection. PMID- 9330473 TI - Bacteriological monitoring of Salmonella enteritidis carrier birds after decontamination using enrofloxacin, competitive exclusion and movement of birds. AB - Two hundred and forty, four-week-old laying birds naturally infected with Salmonella enteritidis PT33 (Pasteur Institute phage typing system) were randomly divided twice (before and during the treatments) to obtain four separately housed groups of 60 birds and to study the efficacy of three decontamination treatments: enrofloxacin either with or without the movement of birds to a clean area, and enrofloxacin combined with movement of birds and a competitive exclusion treatment. The control group remained untreated. In each group contamination with S enteritidis was checked bacteriologically, every week from two months before until two months after the treatments began. All the samples taken from all the birds before the treatments began were S enteritidis-positive. After the treatments it was not possible to isolate salmonella either from the environment or from the faeces of the three treated groups. All the birds were humanely sacrificed at 22 weeks of age and samples of liver, spleen, ovaries and caeca were analysed for the presence of salmonella. The results demonstrated that although antibiotic therapy, the movement of birds into a clean house and competitive exclusion, either combined or not, had some efficacy in reducing infection levels, it was not possible to decontaminate all the birds completely. PMID- 9330474 TI - Value of alpha 1-acid glycoprotein in the diagnosis of feline infectious peritonitis. AB - Feline infectious peritonitis (FIP) is notoriously difficult to differentiate from the many other diseases with similar clinical signs and at present the only conclusive diagnostic test is the histopathological examination of a biopsy. The potential value of raised levels of the acute phase reactants, alpha 1-acid glycoprotein (AGP) and haptoglobin in the diagnosis of the disease was investigated. The concentrations of the two proteins were determined in serum samples from healthy cats and gave reference ranges of 0.1 to 0.48 g/litre and 0.04 to 3.84 g/litre, respectively. Levels of AGP greater than 1.5 g/litre in serum, plasma or effusion samples were found to be of value in distinguishing field cases of FIP from cats with similar clinical signs and differentiated these two groups of cats more effectively than the albumin:globulin ratio. The concentration of haptoglobin was higher in cats with FIP than in the group of healthy cats, but this protein was not of value in the diagnosis of FIP. Serum samples from feline immunodeficiency virus-infected cats were also analysed for these proteins and their concentrations were significantly elevated, illustrating that raised levels of AGP and haptoglobin are not pathognomonic for FIP. PMID- 9330476 TI - Application of the polymerase chain reaction for the routine identification of Mycoplasma bovis. PMID- 9330475 TI - Molecular and clinicopathological diagnosis of malignant catarrhal fever in cattle, deer and buffalo in New Zealand. AB - Fresh and formalin-fixed tissues and blood samples in ethylenediaminetetraacetate were collected from cattle, deer and buffalo with clinical signs suggestive of malignant catarrhal fever (MCF). In addition, formalin-fixed, paraffin-embedded tissue blocks collected from these animals and retrospectively from field cases of MCF were examined. DNA samples extracted from these samples were analysed by polymerase chain reaction (PCR) assay using primers specific for the sheep associated (SA)- and wildebeest-associated (WA)-MCF viruses. Both the SA-MCF virus and WA-MCF virus PCR yielded positive results which were in nearly complete agreement with the histopathological diagnoses of MCF in fresh and formalin fixed, paraffin-embedded tissue from 29 cattle, 24 deer and three buffaloes. Some blood samples tested by the two assays indicated that some of the infected cattle were possible carriers. PMID- 9330477 TI - Examination of red foxes (Vulpes vulpes) from Belgium for antibody to Neospora caninum and Toxoplasma gondii. PMID- 9330478 TI - Confirmation of ovarian remnant syndrome in the queen using hCG administration. PMID- 9330479 TI - BSE and British cattle exports. PMID- 9330480 TI - Preventive role in cattle production. PMID- 9330481 TI - Farmed deer as a potential source of verocytotoxin-producing Escherichia coli O157. PMID- 9330482 TI - Student numbers and university funding. PMID- 9330483 TI - Unilateral uterine swelling in a cow. PMID- 9330484 TI - Lettuce as a suspected cause of narcosis in a duckling. PMID- 9330494 TI - Characteristics of in-patients transferred to a locked ward in a Scottish psychiatric hospital. AB - OBJECTIVE: To identify and characterise in-patients transferred from open wards to the locked facility (IPCU) of a psychiatric teaching hospital, and to classify reasons for transfer. DESIGN: Retrospective case note study using the Lothian Psychiatric Case Register. A reason for transfer classification was devised. SETTING: Intensive Psychiatric Care Unit (IPCU) of the Royal Edinburgh Hospital. SUBJECTS: All in-patients transferred from other wards of the Royal Edinburgh Hospital to the IPCU during one year. RESULTS: Over one year there were 131 transfer episodes involving 97 patients. Ninety-eight per cent of episodes could be allocated to one of six categories of reason for transfer, with physical violence to others being commonest (30%). Patients' mean age was 31 years. There was a predominance of schizophrenics (51%) and males (66%). In 20% of transfers, patients were not detained under mental health legislation. Associations emerged between schizophrenia and physical violence, hypomania/mania and generally disruptive behaviour, personality disorder and self harm. In half of all transfers the patient had already been an in-patient for at least 1 month. Fifty seven per cent of patients were transferred more than once over the preceding five years. Most (90%) transfers originated from acute wards, and patients stayed in the IPCU less than two weeks (70%) before returning to the open wards (23% less than 24 hours). CONCLUSIONS: Local locked wards deal with a range of disturbed behaviour and can be an important component in the spectrum of secure provision. Further evaluative research is required. PMID- 9330495 TI - Psychiatrists and the restricted hospital order in Scotland. PMID- 9330496 TI - Demand for warfarin anticoagulation monitoring in Tayside, Scotland. AB - AIM: The aim of this study was to assess the demand for warfarin prescribing and monitoring, and to identify patients with atrial fibrillation who might benefit from warfarin therapy. The study was carried out in the population of Tayside, Scotland (400,000 people) using patient-specific dispensed prescribing and hospitalisation data from the Medicines Monitoring Unit at the University of Dundee. METHODS: The incidence and prevalence of digoxin and warfarin prescribing were calculated between 1989 and 1993. Patients dispensed digoxin in 1993 were assumed to have atrial fibrillation and they were stratified into high risk groups for an adverse thromboembolic event based on past medical history. The numbers of patients at high risk who were judged to be possible candidates for warfarin were calculated. RESULTS: The prevalence of warfarin prescribing is increasing in Tayside and is mainly for elderly patients. There were also many patients assumed to have atrial fibrillation who were at particularly high risk for an adverse thromboembolic event, who had no record of warfarin prescribing. Only 35% received warfarin. Even given the methodological limitations of this study, and the use of aspirin as an alternative prophylactic agent, it is likely that these patients have been a source of increased prevalence of warfarin prescribing since 1993 and will be in the future. Other indications for warfarin prescribing are also increasing. CONCLUSION: It is anticipated that there will be increasing demands for anticoagulant monitoring, which will need to be met either by increasing the capacity of existing clinics, or by increasing the role of primary care. PMID- 9330497 TI - Can audit of a local protocol for the management of lipid disorders effect and detect a change in clinical practice? AB - OBJECTIVE: To audit the implementation of a local protocol for the management of lipid disorders in Grampian. INTERVENTION: A local protocol for the detection and management of lipid disorders, developed by a multi-disciplinary group and based on available national guidelines and systematic reviews, was disseminated and implemented using the audit cycle. DESIGN: An uncontrolled before and after study using four surveys of different aspects of lipid management in primary and secondary care. RESULTS: There was evidence of change in clinical practice following protocol implementation. Improvements in patient care were identified. However, some areas of inappropriate practice remained unaltered. Changes in dietary therapy indicated an improvement in the appreciation of the importance of appropriate dietary advice. There was an increase in the proportion of patients who were appropriately screened and managed, and appropriately referred to the lipid clinic. The health status of patients being screened/treated following guidelines was poorer suggesting a reduction in screening of low risk patients and an increase in lipid assessment of patients with other health problems or risk factors i.e. more appropriate screening. However, after the protocol was disseminated, general practitioners screened less often for secondary causes of hyperlipidaemia. PMID- 9330498 TI - Attitudes of general practitioners who practice in remote island communities. AB - OBJECTIVE: To describe the personal, social and medical attitudes of doctors who practice on the islands off the West coast of Scotland. DESIGN: Questionnaire survey with a single follow-up. SUBJECTS: All 65 general practitioners (GPs) who practice on the 17 islands located off the West coast of Scotland. RESULTS: Fifty two (80%) responded after a single reminder. The main advantages identified were continuity of care, personal relationships with patients and involvement with the local community. Other important reasons were the opportunity to exercise clinical skills and appreciation of their local environment such as the beauty of the scenery. Disadvantages were associated with isolation, difficulty in obtaining cover, in attending refresher courses and the burden of sole responsibility. CONCLUSION: The GPs who practice in remote island practices believe that the advantages outweigh the disadvantages. They value continuity of care and their relationships with patients and communities. This survey suggests that the difficulties of staffing remote island communities may partly be addressed by allowing undergraduate students and postgraduate colleagues access to these general practitioners. PMID- 9330499 TI - Measuring health-related quality of life outcomes in women with endometriosis- results of the Gynaecology Audit Project in Scotland. AB - The clinical management of endometriosis was addressed within the recent Gynaecology Audit Project in Scotland. The impact of endometriosis and its treatment on women's health-related quality of life was examined using a condition-specific measure and a general measure, the Short Form 36 health survey (SF-36). Postal questionnaires containing the health-related quality of life measures were sent to 273 women at diagnosis and six months later. The measurement properties, including the reliability, validity and responsiveness, of the measures were examined. The condition-specific questions and the SF-36 had a high level of reliability. The validity of the condition-specific scores was demonstrated by their high correlation with the SF-36 which is a well-validated measure. Furthermore, the condition-specific scores were related to clinicians' assessment of disease severity and the need for further treatment. At the six month follow-up, changes in scores conformed to expected hypotheses, demonstrating the responsiveness of both measures. As a general measure, the SF 36 appeared to reflect the effects of both the condition of interest (i.e. endometriosis) and other conditions affecting health at the time of measurement (i.e. treatment side effects). The condition-specific measure was more responsive than the SF-36 to the changes in pain symptoms which resulted from active treatment. A condition-specific questionnaire, together with a general measure such as the SF-36 health survey, can provide a reliable, valid and responsive package of measures for assessing health-related quality of life in women with endometriosis. Such measures should be used alongside clinical measures of outcome to assess the effectiveness of different treatment strategies for endometriosis. A similar approach combining general and specific instruments would be useful in medical audits of other conditions. PMID- 9330500 TI - A review of school vision screening in Glasgow. AB - OBJECTIVES: To assess to what extent the primary school vision screening programme was being carried out in the years 1993-94 and 1994-95, what the prevalence of eye defects was in the primary schools and how many children had a positive screening test. DESIGN: Summary sheets of the results of screening were returned by each school nurse. SETTING: All 315 mainstream primary schools served by Greater Glasgow Health Board. SUBJECTS: All school nurses working in these schools. RESULTS: Large numbers of tests were being performed but the screening programme was not fully carried out in 10% of schools in 1993-94 and in 68% in 1994-95, although the number of schools where some screening was performed did increase. The years were not directly comparable due to a change in the age groups screened. When a class was screened 94-96% of children were tested. The rate of positive screening varied from 4% to 15.6% and varied with age and locality. Fewer abnormalities were found at five years than in the older age groups (Primary 1-7.6%, Primary 4-11.4%, and Primary 7-9.7%) and fewer abnormalities were detected in the North-West area where a pre-school orthoptic screening programme was operating (4% in the North-West compared with 11% in the South where no pre-school screening was carried out). CONCLUSION: If the recommended programme is carried out, then 95% of children at age five years could be tested at a stage when amblyopia is still potentially treatable. The reasons for the programme not being completed require to be addressed. Variations between areas need to be confirmed and the reasons for them identified. Further evaluation of the programme is required, exploring the accuracy of it and the long-term outcomes. PMID- 9330502 TI - Death in the PICU: caring for the "other" families. AB - This article discusses the care of Pediatric Intensive Care Unit (PICU) families who are present during the death of a child other than their own. "Other" families witness bereavement and experience grief, and there is no literature available to evaluate and describe their needs. Many PICU families are exposed to the death of a child. Nursing interventions to ameliorate self-awareness skills, validate the significance of the event, eliminate unnecessary fears, and cultivate effective coping strategies are necessary to decrease the stress of the experience. Research is needed to specify the needs of "other" families and to improve care for everyone present during the death of a child. PMID- 9330501 TI - Risk factors for cardiovascular disease in children with type I diabetes: Part 1. AB - Diabetes is a major risk factor for premature morbidity and mortality caused by cardiovascular disease (CVD). Although an increased prevalence of lipid abnormalities in many populations with diabetes has been observed, minimal data exist regarding the distribution, correlates, and determinants of lipid levels of children with diabetes. Early identification of hyperlipidemia and other CVD risk factors is requisite to timely and specific nursing interventions. Part 1 of this two part series will discuss the lipid profile, the link between cardiovascular disease and diabetes, and physiological risk factors for CVD in children with diabetes. PMID- 9330504 TI - Coordinated systems of community-based health care delivery: a vehicle for health care reform. AB - The need for health care reform in this country continues to be high on the public agenda. Deficits in the access of United States citizens to needed health care services continues, despite recent efforts at health care reform. In addition, the health status of children in this country is of paramount concern. Significant threats to the health of this nation's children have spurred several proposals for reducing the cost of health care while improving the quality of outcomes. A sample of these proposals is analyzed. An example of a health care program which exemplifies one such proposal for health care reform is then discussed. Findings of a recent research project that is relative to the benefits of such a program are provided. PMID- 9330503 TI - Reconstructing reality: family strategies for managing childhood cancer. AB - The purpose was to describe strategies used by the family in response to childhood cancer and to relate those strategies to two different conceptual frameworks. A longitudinal, prospective, grounded theory study was conducted with a sample of 32 members of seven families who had a child recently diagnosed with cancer. All family members 5 years and older participated in three semistructured home interviews. Constant comparative analysis was used. The core process in which families engaged was reconstructing reality, using strategies of managing the flow of information, reorganizing roles, evaluating and shifting priorities, changing the future orientation, assigning meaning to the illness, and managing the therapeutic regimen. These strategies support a Family Management Style framework for viewing family management of pediatric cancer. PMID- 9330505 TI - African-American families with chronically ill children: oversights and insights. AB - From a critical review of the literature concerning African-American families' management and care of children having chronic illness, we concluded that information on culture-related experiences in such families remains seriously deficient. To present an accurate picture of African-American life as these families manage a child with a chronic illness, more comprehensive and detailed descriptions of family caregiving styles and other experiences are needed. PMID- 9330506 TI - Health insurance for poor children. PMID- 9330507 TI - Using equipment in unfamiliar clinical settings: audiology screening. PMID- 9330508 TI - Sibling relationships of Japanese children with diabetes. AB - Sibling relationships among diabetic children and their sibling are similar to those among healthy children. However, there was more sibling interaction among diabetics and their siblings than among healthy children. The diabetic's siblings are interested in learning about diabetes and in being supportive to their ill brother or sister. However, the feelings are different among the ill children. The ill sibling does not feel that the healthy sibling is supportive of the strict dietary regimen. This is especially true when it comes to eating sweets in front of them. Many parents viewed their healthy children as supportive to the patients, and they wished them to continue to be supportive persons to the patients in the future. Some parents felt guilt and conflict about spending more time with the ill child. PMID- 9330509 TI - [Surgery of aortic arch aneurysm combined with other cardiovascular lesion]. AB - From 1995 to 1996, we performed aortic arch replacement using antegrade cerebral perfusion under deep hypothermia in 7 patients, in whom 4 cases accompanied with cardiac lesion which treated simultaneously and 3 cases had abdominal aortic aneurysm. We compared the surgical results between cases with (group II, n = 4) and without (group I, n = 3) combined cardiovascular lesion. There is no difference between two groups in the cerebral perfusion time and the amount of bleeding and blood transfusion. The cardiac ischemic time and bypass time were insignificantly longer in group II than in group I. We experienced no early death and no cardiac and brain complication in both groups. Three cases with abdominal aortic aneurysm had two-staged operation successfully after arch surgery within a half year. In conclusion, we successfully treated aortic arch aneurysm even in patients combined with other cardiovascular lesion as well as in patients without that. PMID- 9330510 TI - [Composite arterial conduits with internal thoracic artery and radial artery for a myocardial revascularization]. AB - From April 1996 to December 1996, 15 patients were submitted to myocardial revascularization using composite arterial conduit with internal thoracic artery (ITA) and radial artery (RA). The age ranged from 51 to 76 years (mean age, 65.7 years); Forty patients were male. All patients had double or triple vessel disease or LMT disease. We used 28 arterial conduits including 15 left ITAs, 15 RA, 9 right gastroepiploic artery and one inferior epigastric artery. 15 RAs were anastomosed to LITAs and 15 composite arterial conduits were constructed (branched in 15). There was no operative deaths. Early postoperative angiographic controls demonstrated 93.3% (14/15) patency of composite grafts in 14 of 15 patients. The composite arterial graft using ITA and RA is feasible and the anastomoses so performed are completely safe. PMID- 9330511 TI - [Successful treatment of myocardial infarction of left main trunk by emergent CABG under IABP and PCPS support]. AB - We experienced a successful treatment of acute myocardial infarction which was due to left main trunk obstruction. A 54-year-old man with no history of angina was transported by a rescue squad in cardiogenic shock, and diagnosed by electrocardiography with a wide range of myocardial infarction. Emergent coronary arteriography was performed under IABP support, revealing 99% stenosis in the left main trunk. Percutaneous transluminal coronary recanalization (PTCR) was performed, but suddenly cardiac arrest was happened. He was put on emergency percutaneous cardiopulmonary support (PCPS). A Palmaz-Schatz stent was implanted for reperfusion, but the patient was hemodynamically unstable with frequent ventricular arrhythmia and pulmonary edema. 24 hours later he underwent coronary artery bypass grafting and CPB could be terminated intraoperatively. His cardiac function was very low and LVEF was 20%. All grafts were patent. On the rehabilitation he was discharged on postoperative day 162 and has returned to work in his office one year postoperatively. PMID- 9330512 TI - [Thoracoscopic unilateral fold plication method in lung volume reduction surgery for diffuse pulmonary emphysema]. AB - We have devised a 'Fold Plication Method', which helps ensure safe and simple lung volume reduction surgery (LVRS) for pulmonary emphysema and prevent air leakage from pulmonary stumps, which is most crucial problem, in addition to postoperative interstitial pneumonia that is occasionally caused by the use of the bovine pericardium. We performed LVRS on seven patients with the disease, using this method and the thoracoscopic surgery, which is based on the 'Two Windows Method' we had previously developed. The operation was performed without observable bleeding within approximately one hour for all patients, who then recovered favorably. PMID- 9330513 TI - [Surgical results of emergency coronary artery bypass grafting: effect of left main trunk lesion]. AB - Emergency coronary artery bypass grafting has higher risk than elective surgery. Furthermore, if a lesion is located at left main coronary artery, the frequency of cardiogenic shock or high risk patient would be greater, and operative results would be worse. Between January, 1989 and December, 1995, 45 patients who underwent emergency CABG were included. Age ranged 44 to 80 years (mean 67 +/- 7.6 years; 31 men, 14 women). Of 45 cases, 12 cases were patients with LMT lesion. Results were analysed by univariate analysis and multivariate logistic analysis. Of 45 emergency cases, 5 were operative death and 3 were hospital death. Mortality rate was 17.8%, which was significantly higher than the mortality of elective CABG (2.8%) during the same period (p < 0.001). A factor that influenced the mortality was acute myocardial infarction (AMI), which was confirmed by both univariate and multivariate logistic analyses. Odds ratio (relative risk) was 12.4 for AMI. Only one patient died in 12 cases with LMT (8.3%). This case was due to complication after catheter intervention possibly caused by MOF. Thus, the relative risk of LMT was not so high (p = n.s). Other factors showed no significant correlation. Although the emergency case of LMT lesion was generally severe, we could have the same result as other emergency surgeries when patients were revascularized as soon as possible. PMID- 9330514 TI - [Effects of milrinone in patients undergoing cardiac surgery]. AB - We investigated effects of milrinone in twenty consecutive patients (6 adults, 1 child, and 3 early infants) during cardiac surgery requiring cardiopulmonary bypass (CPB). The operations were: CABG 5, CABG+mitral valve repair 2, MVR 2, redo-MVR 4, aortic surgery 3 (total arch replacement 2), VSD+pulmonary hypertension 2 (infants), Tetralogy of Fallot 1, and PDA aneurysm 1 (infant). Ten minutes after release of aortic cross-clamp, all patients received milrinone by loading dose (50 micrograms/kg, bolus), followed by a continuous infusion of 0.5 or 0.75 microgram/kg/min. All of patients weaned from CPB with milrinone and low dose of dopamine. The 75% of patients did not require any other drugs except for milrinone during post operative ICU stay. At the same time, we evaluated the effect of milrinone on platelet number in the patients before and after CPB. Milrinone administration did not cause significant changes in platelet number after CPB. No adverse effects attributable to this drug were found. Milrinone appears to be effective and safe in patients undergoing cardiac surgery of all kinds. PMID- 9330515 TI - [A study on low grade malignant tumors arisen in the trachea and the bronchus]. AB - Twelve patients who suffered from low grade malignant tumors arisen in the trachea and the bronchus (6 of carcinoid, 4 of mucoepidermoid carcinoma, and 2 of adenoid cystic carcinoma) underwent surgical treatment from 1977 to 1996 in our department. Operations included 1 sleeve resection of the trachea, 1 patch plasty of the trachea, 3 sleeve lobectomies, and 7 lobectomies. Lymph node dissection was performed in 9 of 12 cases. Metastases in lymph nodes were not found in all 12 cases. Five year survival rate of low grade malignant tumors arisen in the trachea and the bronchus was 78.8% and better than that of stage I lung cancers. PMID- 9330516 TI - [A case report of total removal of infected pacemaker leads with cardiopulmonary bypass through right thoracotomy]. AB - A 61-year-old man with septicemia had four infected pacemaker leads, which were impossible to remove using simple traction method. He received CABG previously, and SVG anastomosed to LAD was patent. Redo median sternotomy had a possibility to make damage to SVG. Total removal of infected pacemaker was performed successfully with cardiopulmonary bypass through right thoracotomy. PMID- 9330517 TI - [Thoracoscopic surgery for the giant bulla complicating pulmonary eosinophilic granuloma: a case report]. AB - We report a case of pulmonary eosinophilic granuloma (PEG) accompanied with a giant bulla successfully operated with a thoracoscopy. A 24-year-old female had been suffering from recurrent bilateral pneumothoraces with multiple bullae since February 1993. She was diagnosed as PEG with an open lung biopsy in July 1994. Repetitive pleurodesis were efficacious against the pneumothoraces. However, a bulla in the left upper lobe were growing with her respiratory function having worsened to bed rest. In May 1995, thoracoscopic surgery were performed to extinguish the bulla. Trocars were directly introduced into the lumen of the bulla. Ligation of the three communicating bronchial branches and tube drainage made the bulla disappear immediately. The patient had been well for one and a half year postoperatively. PMID- 9330518 TI - [Findings of the Carpentier-Edwards porcine bioprosthesis in the mitral position with PTF in the 16th post-operative year]. AB - A 62-year-old woman, who had ever undergone mitral valve replacement with a 29 mm Carpentier-Edwards (C-E) porcine bioprosthesis for mitral regurgitation, was admitted to our hospital because of progressive dyspnea on effort. Transthoracic and transesophageal echocardiography revealed primary tissue failure (PTF) of the C-E bioprosthesis with prolapsing, thickening and impaired mobility of the leaflets which resulted in severe mitral stenosis and regurgitation. The patient was scheduled to elective operation for redo mitral valve replacement with a prosthetic mechanical valve. The removed C-E bioprosthesis showed (1) leaflet perforation, (2) commissural tear, (3) pannus overgrowth, (4) impaired leaflet mobility and (5) diffuse calcification. Despite all attempt to improve on bioprosthetic features, reported free rate from PTF decreased approximately after 10th postoperative year in any types of bioprostheses available. Our various degenerative findings on the C-E bioprosthesis in the 16th postoperative year may suggest the limitation of long-term durability of the bioprostheses, which needs to be concerned about in their surgical indication and choice. PMID- 9330519 TI - [A case of two stage operation of thoracic and thoracoabdominal aortic aneurysm of a patient in late phase with Behcet's disease]. AB - Patient was a 70-year-old female diagnosed as Behcet's disease 1973. She underwent descending thoracic aortic aneurysm operation 1980. She had graft replacement for a residual thoracoabdominal aortic aneurysm on April 28, 1994. She didn't accept steroid therapy although she had a slight inflammatory reaction on admission. The graft replacement of thoracoabdominal aortic aneurysm was done with an aid of Carmeda closed chest support system bypass and segmental aortic cross clamping. Reattachment of the intercostal and lumbar arteries to the graft was used with button technique or interposition technique. Visceral branches including celiac axis, superior mesenteric and right renal arteries were reconstructed to an opening made in the graft with button technique and reattachment of left renal artery was used with graft interposition during selective perfusion of visceral arteries. She started steroid therapy with 5 mg of predonine to prevent inflammatory reaction postoperatively. The thoracoabdominal aortic aneurysm accompanied with Behcet's disease was relatively rare and this is a long survival case. PMID- 9330520 TI - [Case report of successful treatment of patent ductus arteriosus with severe pulmonary hypertension in an adult]. AB - A 40-year-old man was diagnosed as having PDA with severe PH by cardiac catheterization. We decided that surgery was indicated on the basis of 100% oxygen inhalation test. After 100% oxygen inhalation, left to right shunt changed from 37.8% to 61.5%, Qp/Qs changed from 1.43 to 2.60, and PVR changed from 1,199 dyne/sec/cm5 to 237 dyne/sec/cm5. We decided to perform surgery. Under extracorporeal circulation, PDA was safely closed by triple ligation and was directly closed after pulmonary arteriotomy. The postoperative course was uneventful. Severe PH was greatly improved, pulmonary artery pressure recovering to the normal range, by administering PGE1, prostacyclin (PGI2), and nitroglycerin. PMID- 9330521 TI - [Endoscopic transthoracic sympathectomy for angina pectoris: a case report]. AB - We performed endoscopic transthoracic sympathicotomy (ETS) for angina pectoris in a 77-year-old female. She had multiple coronary disease of a history of failed percutaneous transluminal coronary angioplasty (PTCA). Bilateral ETS of Th2 to Th5 was carried out under general anesthesia without surgical complication, hemodynamic change and ECG change. Postoperatively she was free from angina and increased working capacity. ST depression on ECG was remarkably improved and the increase of heart rate and systolic blood pressure was suppressed on exercise test after a month of ETS. The effect of ETS to angina pectoris was suspected not to relieve anginal pain but also to reduce myocardial oxygen demand on exercise. We conclude that ETS is a safe, simple and low invasive procedure and it can be done in patients who are judged unsuitable for coronary artery bypass grafting (CABG) or PTCA. PMID- 9330522 TI - [A case of permanent pacemaker implantation in neonate using steroid-eluting myocardial pacing leads]. AB - A 0-day-old infant with symptomatic congenital atrioventricular block diagnosed by fetal echocardiography was successfully treated with the pacemaker implantation. Two steroid-eluting myocardial electrodes were fixed on the wall of the right ventricle. Steroid-eluting myocardial electrode is smaller than the others, stab-in and screw-in leads, so even in neonate we were able to place the leads by the subxyphoid approach. The postoperative thresholds were excellent. PMID- 9330523 TI - [Adult bochdalek hernia after playing at a tug of war]. AB - A 38-year-old female was admitted to Shonai Hospital with severe abdominal pain and nausea after playing at a tug of war in the athletic meeting. The X-ray film showed air above the left diaphragm, and CT scan and barium enema revealed the incarcerated transverse colon to the left thoracic cavity. Operation was performed through a thoracotomy. Because of no evidence of trauma, the case was diagnosed as adult Bochdalek hernia. Repair could be done by direct suture and her postoperative course was uneventful. PMID- 9330525 TI - [A case of thymic cyst and thymoma with myasthenia gravis]. AB - A case of 53-year-old male with thymic cyst was reported. We established diagnosis for myasthenia gravis. The chest CT and MRI demonstrated a cystic and mass lesion on the anterosuperior mediastinum. This finding suggested a thymic cyst and tumor with myasthenia gravis. The extended thymectomy was performed. Histological examination demonstrated the clear separation of the mixed type thymoma and the thymic cyst. The few reported cases of thymic cyst and thymoma with myasthenia gravis were reviewed. PMID- 9330524 TI - [Direct approach to the site of injury of the thoracic duct in treatment of chylothorax after pulmonary resection]. AB - Chylothorax, a rare complication after pulmonary resection, has no single established treatment. Generally, conservative therapy is tried first, but surgery should be done without delay if chyle leakage is severe. A 73-year-old woman underwent upper left lobectomy for lung cancer. Two days later, chylothorax was diagnosed, and because chyle leakage was great, emergency re-thoracotomy was done on day 4 after the first operation. With the preoperative ingestion of 200 ml of milk and 20 g of margarine, chyle leakage from the injured thoracic duct was readily located in the upper mediastinum. Closure of the trunk of the thoracic duct may be undertaken, but a direct approach to the site of injury is more preferable, because the trunk may have collateral. PMID- 9330526 TI - [A case of lung abscess with elevated serum levers of sialyl Lewis X-i (SLX) and CA 19-9]. AB - A case of lung abscess of the felt lower lobe in a 19-year-old woman with elevated serum levels of Sialyl Lewis X-i (SLX) and CA 19-9 is reported. Completing the lobectomy, serum SLX level returned to the normal range within a week postoperative day. Serum CA 19-9 level also decreased at half life of 2 weeks to the normal range within 6 weeks postoperative day. Laboratory examination demonstrated high levels of these antigens in abscess fluid. Histologically, the abscess was revealed to be associated with a markedly dilated bronchus with hyperplastic bronchial glands, and there was no evidence of malignancy. Immunohistochemical examinations using monoclonal antibodies against human SLX and CA 19-9 showed highly positive reaction with those antigens in both goblet cells in bronchial epithelia and the mucous cells in bronchial glands. PMID- 9330527 TI - [Successful surgical treatment for type B aortic dissection with Marfan's syndrome after aortic root and mitral valve replacement: report of a case]. AB - A 30-year-old female with Marfan's syndrome underwent aortic root replacement for annuloaortic ectasia and mitral valve replacement for mitral regurgitation. She remained well until 16 months postoperatively when she had sudden onset of pain. Preoperative angiogram showed Stanford B aortic dissection. Thoracoabdominal aortic replacement was performed successfully under deep hypothermic bypass. PMID- 9330528 TI - Induction of resistance to etoposide and adriamycin in a human glioma cell line treated with antisense oligodeoxynucleotide complementary to the messenger ribonucleic acid of deoxyribonucleic acid topoisomerase II alpha. AB - Acquisition of resistance to anticancer agents is a serious problem for cancer chemotherapy. The present study analyzed the relationship between expression of the alpha isoform of deoxyribonucleic acid (DNA) topoisomerase II (topo II alpha) and chemosensitivity to topo II inhibitors by modulating the level of topo II alpha expression. A phosphorothioate analogue of an 18-nucleotide oligomer which is complementary to the translation initiation site of the human topo II alpha messenger ribonucleic acid sequence was used to suppress the expression of topo II alpha in a human glioma cell line (U373MG). The topo II alpha activity of the treated cells was reduced to 1/3 of untreated cells in a decatenation assay using kinetoplast DNA. Antisense oligoDNA-treated cells showed mild resistance to the topo II inhibitors, etoposide and adriamycin, of about 2.0 fold and 1.5 fold, respectively, compared to control cells. Only partial reduction in the activity of topo II alpha in the glioma cell line can cause a measurable resistance to topo II inhibitors, implying that the degree of topo II expression is correlated with chemosensitivity to topo II inhibitors. PMID- 9330529 TI - Changes in brain glutamine synthetase activity in congenital hydrocephalic rats (LEW-HYR) after ventriculoperitoneal shunt. AB - Significantly reduced activities of glutamine synthetase (GS), which is predominantly present in glial cells, occur in the early stage of congenital hydrocephalic rat (LEW-HYR) brain development. GS activity is reported to be related to brain dysfunction. The effect of ventriculoperitoneal (VP) shunt on the suppression of GS activity was studied in the LEW-HYR. VP shunting improved the attenuation of GS activity in the LEW-HYR and the response of GS activity to methionine sulfoximine (a competitive GS inhibitor) treatment was similar to that seen in normal siblings. However, no enhancement of GS activity by hydrocortisone could be detected, although this enhancement occurs in the normal siblings. These results suggest that VP shunting is not completely effective in improving the suppression of brain GS activity in the LEW-HYR, since the suppression of GS activity in the LEW-HYR might be programmed genetically. PMID- 9330530 TI - Intracerebral monoamine concentration after ventriculoperitoneal shunting in the congenital hydrocephalus rat. AB - This study investigated the relationship between neurotransmitters and improvement of symptoms after ventriculoperitoneal shunting in congenital hydrocephalus (LEW-HYR) rats. Twenty-four patent hydrocephalus rats, aged 12-14 days, were randomly assigned to the following four groups: ventriculoperitoneal shunt group, obstructed shunt group, burr hole group, and no treatment group. In addition, six normal rats served as control group. Head to body length ratio was measured before and 7 days after the procedures. Coordination movement was evaluated on the 7th postoperative day. High performance liquid chromatography (HPLC) was used to measure the concentrations of dopamine (DA), norepinephrine (NE), serotonin, homovanillic acid (HVA), 3-methoxyl-4-hydroxyphenylenglycol, 5 hydroxy-indolacetic acid (5-HIAA), and 3,4-dihydroxyphenylacetic acid in the whole cerebral cortex, the thalamus and hypothalamus, the midbrain, the lower brainstem, the cerebellum, and the striatum. Fluorohistochemical studies were also performed. Significant improvements were observed in body proportion and coordination movement in the ventriculoperitoneal shunt group compared to the burr hole group and the no treatment group. HPLC and fluorohistochemical studies revealed that concentrations of NE in the thalamus and hypothalamus and DA in the striatum were significantly lower in the burr hole group and the no treatment group. Concentrations of HVA and 5-HIAA in the cerebellum were significantly lower in the control group. The present study indicates that ventriculoperitoneal shunting may improve the changes in concentrations of neurotransmitter in specific neurons caused by hydrocephalus, and this may contribute to the improvement of the symptoms. PMID- 9330531 TI - Iatrogenic arteriovenous fistula of the middle meningeal artery caused during embolization for meningioma--case report. AB - A 73-year-old female developed middle meningeal arteriovenous fistula during embolization of a falx meningioma. The cause of this complication was thought to be perforation by the guide wire during catheterization at the sharp bend in the sphenoidal portion of the middle meningeal artery. Embolization of the fistula and the feeding artery to the meningioma with polyvinyl alcohol particles 250-355 microns size resulted in complete obliteration of the fistula. Computed tomography showed no epidural or subdural hematoma. Introduction of the microcatheter beyond the sharp bend in the middle meningeal artery should not be attempted to avoid the possibility of iatrogenic middle meningeal arteriovenous fistula. PMID- 9330532 TI - Endovascular treatment of a partially thrombosed giant basilar tip aneurysm using interlocking detachable coils--case report. AB - A 65-year-old female presented with visual acuity loss. Magnetic resonance imaging confirmed the presence of a partially thrombosed giant aneurysm on the basilar tip. Cerebral angiography showed the opacified lumen of the aneurysm was 25 x 15 mm with a broad-based neck. Using a transfemoral approach, a microcatheter was guided through the vertebral artery and placed directly into the aneurysm under local anesthesia. Interlocking detachable coils were deposited into the patent portion of the aneurysm, resulting in 95% obliteration of the aneurysm and preservation of the parent artery. No complication was observed during or after surgery. Follow-up angiography 2 months later demonstrated the aneurysm was 95% occluded. No coil compaction was observed. Endovascular coil embolization therapy provides a therapeutic option for management of basilar tip aneurysms. PMID- 9330533 TI - Primary choroid plexus papilloma of the foramen magnum--case report. AB - A 50-year-old male presented with a choroid plexus papilloma in the foramen magnum manifesting as dysesthesia in the right hand and severe headache. Magnetic resonance imaging clearly showed that the tumor was located in the cerebellomedullary cistern, without extension into the fourth ventricle. However, differentiation from hemangioblastoma or foramen magnum tumor was difficult by neuroimaging. Intraoperative observation found the tumor was located extraventricularly and attached to the choroid plexus of the foramen of Magendie. The tumor was grossly totally resected. Histological examination proved the tumor was a choroid plexus papilloma without malignancy. His neurological deficits resolved almost completely. PMID- 9330534 TI - Orbital hemangiopericytoma--case report. AB - A 43-year-old male presented with swelling involving the right eye. T1-weighted magnetic resonance imaging demonstrated a round tumor in the lateral region of the right orbit, which was isointense relative to the cerebral gray matter and homogeneously enhanced by gadolinium-diethylenetriaminepenta-acetic acid with a flow-void signal area in the mass. The tumor was totally resected through the transcranial and fronto-orbitotemporal approach after embolization of feeding arteries arising from the external carotid artery. The histological findings were characteristic of hemangiopericytoma. No radiation therapy was administered. The transcranial and fronto-orbitotemporal approach provides a wide operative field with excellent exposure of the highly vascular orbital tumor. PMID- 9330535 TI - Recurrent Lhermitte-Duclos disease--case report. AB - A 43-year-old male presented with recurrent Lhermitte-Duclos disease (LDD), a rare pathological entity of the cerebellum of which the etiology is still controversial. He had undergone subtotal removal of a cerebellar lesion, misdiagnosed as a benign astrocytoma, 8 years previously. Subtotal removal of the recurrent tumor completely resolved the presenting symptoms. Recurrence of LDD is not as rare as generally assumed. Patients with LDD require long-term observation even when the initial treatment appeared curative. PMID- 9330536 TI - Lhermitte-Duclos disease associated with Cowden's disease--case report. AB - A 49-year-old Japanese male with Lhermitte-Duclos disease subsequently developed a very rare association with Cowden's disease. Partial tumor removal established the diagnosis of Lhermitte-Duclos disease. Follow-up examinations discovered the presence of Cowden's disease. Long-term follow-up of patients with Lhermitte Duclos disease is essential to identify signs of Cowden's disease, which carries the risk of developing malignancy. PMID- 9330537 TI - Unilateral sensori-neural hearing disturbance caused by intramedullary cerebellar tumors--three case reports. AB - Three patients presented with unilateral sensori-neural hearing disturbance as the initial symptom of cerebellar tumors: a 19-year-old female with a medulloblastoma (Case 1), a 45-year-old male with a cerebellar low-grade glioma (Case 2), and a 49-year-old female with a cerebellaer hemangioblastoma (Case 3). In Cases 1 and 2, the whole length of the eight cranial nerve was intact according to magnetic resonance imaging and intraoperative findings. In Case 3, the intracerebellar tumor had bulged into the cerebellopontine cistern, compressing the eighth cranial nerve near the brainstem. Auditory evoked brainstem responses showed only the first wave in all three patients, and the following waves could not be discriminated. Unilateral sensori-neural hearing disturbance occurs very rarely in patients with intramedullary cerebellar lesions because the auditory neural pathway is bilaterally innervated. Intramedullary tumors may cause unilateral sensori-neural hearing disturbance by infiltrating or causing edematous changes of the eighth cranial nerve or the cochlear nucleus in the brainstem, or by compressing the nerve in the cistern. The symptoms are the same as those of acoustic neurinoma, so intramedullary cerebellar tumors should be considered in the differential diagnosis of unilateral sensorineural hearing disturbance. PMID- 9330539 TI - Active compliance in robotic surgery--the use of force control as a dynamic constraint. AB - Robotic surgery can be carried out automatically by using a robot to move the cutting tool under position control. However, although the surgeon can observe the procedure on a visual display and has the ability to stop the operation in an emergency, he has little direct contact with the task. An alternative approach is to involve the surgeon more directly, by his moving a robot using active force control. The robot is then used to allow motion in preprogrammed regions, by the surgeon back-driving the robot motors, while preventing motion in prohibited areas. This active constraint robot (or ACROBOT) is described in this paper applied to knee surgery, in which the knee bones are accurately machined to allow the fitting of prosthetic knee implants. The ACROBOT is, however, ideally suited to a range of surgical procedure, because it allows the surgeon to feel the forces exerted during cutting and take appropriate action. This ability to be in direct control, while being constrained to cut within a permitted region, enhances safety and makes the system more acceptable to the medical community. The system of programmable constraint also allows the ACROBOT to provide the traditional benefits of robot surgery, namely the ability to machine complex geometrical surfaces very accurately and to make repetitive motions tirelessly. The system also has a potential for minimally invasive procedures. In knee surgery, for example, the robot could operate through a small incision in the skin and excise a volume into which a small, specially designed, unicompartmental prosthesis could fit. PMID- 9330538 TI - Intraoperative guidance in maxillofacial and craniofacial surgery. AB - The authors' experiences with intraoperative computer assisted guidance in interventions in oromaxillofacial and craniofacial surgery are reported. The guidance system SPOCS (Surgical Planning and Orientation Computer Systems, Aesculap, Germany) consists of an infrared light emitting system of diodes and camera, an imaging workstation and assorted freehand instruments. The software is an updated version of the well-known Viewing Wand software (ISG Technologies, Canada). In tests on phantoms, the system proved a mean accuracy of less than 1.5 mm. Within the last 15 clinical tests, the system has achieved an accuracy better than 3 mm which, at the moment, the authors estimate to be sufficient to proceed with its clinical evaluation. Using bone screws to register the patient's position, an accuracy in the range of less than 2 mm in relation to bony reference points has been achieved. By visualizing the tip of the instrument in real time, this technique allows surgical interventions, even in anatomically complicated situations, without endangering vital neighbouring structures. The 'offset' function of the software, by which the surgeon can elongate the tip of the instrument virtually, allows the surgeon to analyse structures before they are penetrated by the instrument as in a 'look ahead' operation. The authors expect computer assisted simulation and guidance systems to improve surgical quality and reduce the risks associated with surgical interventions. PMID- 9330540 TI - A computer assisted orthopaedic surgical system for distal locking of intramedullary nails. AB - This paper presents a prototype system for computer assisted surgery, the purpose of which is to assist orthopaedic surgeons when performing distal locking of intramedullary nails. This system comprises three components, namely: an Intelligent Image Intensifier, a Trajectory Tactician and an Intelligent Trajectory Guide. The Intelligent Image Intensifier is an X-ray vision system that provides accurate X-ray images. Such images enable the Trajectory Tactician software to analyse the operation site and calculate the trajectory required for a screw to lock an intramedullary nail. This involves the capture of two X-ray images from which are extracted the projections of the nail's edge boundaries and its distal locking holes. Using an analytical mathematical model of the nail, the position and orientation of the nail is determined. The trajectory is then implemented by the surgeon using the Intelligent Trajectory Guide. Evaluation in the laboratory suggests that the system is capable of reliably inserting a locking screw into an intramedullary nail. The rapidity with which this computer assisted method achieves locking should benefit both patient and surgeon by reducing radiation dosage and the length of time required to lock a nail. PMID- 9330541 TI - A new oscillating saw for robotic aided surgery. AB - In this paper a brief description of a computer and robotic aided surgery system is given with a detailed overview of the necessity to develop special tools for robotic surgery. The application range of this robotic system has been specially focused on the orthopaedics field and, more particularly, on the execution of osteotomies. It was therefore necessary to develop a new saw device which would meet medical and--from the robot system point of view--mechanical as well as functional requirements. After describing the device which was developed on the basis of these requirements, a detailed comparative study of off-the-shelf oscillating saws and the new device is given at the end of the paper. PMID- 9330542 TI - A technique for measuring contact force distribution in minimally invasive surgical procedures. AB - This paper investigates new methods for measuring forces and tactile sense as a contribution towards relaying the sense of touch to the surgeon. The approach used is to determine a distribution of contact force using a small number of sensory outputs to detect the bending of a surface of known behaviour. Software algorithms have been produced to interpret the contacting force from sensory data, and have achieved a bandwidth of 30 Hz and an accuracy of 2 per cent. The sensor construction is of sufficiently low cost to produce a disposable unit and uses materials that are compatible with the invasive working environment. PMID- 9330543 TI - The Probot--an active robot for prostate resection. AB - As men age, their prostates can enlarge, causing urinary difficulty. Surgery to correct this [transurethral resection of the prostate (TURP)] is a skilled and time-consuming operation requiring many repetitive motions of a cutter. A robot has been developed to perform these motions, relieving the surgeon of much of the burden of surgery. This robot has been tried both in the laboratory and later on human subjects and has proved itself capable of performing prostate resection. The Probot system consists of on-line imaging and three-dimensional prostate model construction, an appropriate surgeon-computer interface, a counterbalanced mounting frame and a computer controlled robot. PMID- 9330544 TI - A non-invasive patient registration and reference system for interactive intraoperative localization in intranasal sinus surgery. AB - A basic problem common to all systems for computer assisted surgery (CAS) is patient referencing, or the transfer of preoperative image data to the intraoperative pathology. The authors describe a highly precise CAS system with non-invasive referencing that can be used in ear, nose and throat (ENT) surgery of the paranasal sinuses. It is based on optical digitizing with several custom made self-localizing surgical instruments. The accuracy of the system was tested in an experimental model using a plastic head. Measurements of repositioning the reference bow had a mean error of 0.81 mm +/- 0.31 mm. The system was evaluated clinically with 11 patients who received surgery for different pathologies of the paranasal sinuses. These trials met with a high rate of success and specific results are reported. PMID- 9330545 TI - Simulation of resistance forces acting on surgical needles. AB - High precision in the manual control of needles and biopsy probes in medical treatment requires high skill and dexterity levels. In anaesthesia, force sensation is an important feedback mechanism, and the practitioner needs to refresh or develop skills to improve on the interpretation of needle progress towards the target site. This paper describes an experimental tactile force simulator for uniaxial needle action for which the force resisting progress of the needle is derived from measured data. As an example, the approach taken to develop the simulation of the insertion of epidural needles is described. Adaptation to other procedures would be possible by adopting new reference models based on appropriate measured force data. PMID- 9330546 TI - Assessing Health-Related Quality of Life in Early HIV Disease. Proceedings of a meeting. Paris, France, 1-2 October 1996. PMID- 9330547 TI - Current antiretroviral therapy: an overview. AB - Advances in the understanding of human immunodeficiency virus (HIV) pathogenesis, clinical assessment with viral load testing and the availability of potent combination antiretroviral therapy regimens have led to significant changes in options for HIV-infected patients. From the first approved antiretroviral agent, zidovudine (AZT), through two-drug nucleoside analogue regimens, to the current three-drug combination regimens with protease inhibitors, both the benefits of therapy and the complexities of therapy continue to increase. With the clinical benefits come associated lifestyle constraints and, thus, the impact and assessment of potent antiretroviral therapy on patient quality of life (QoL) becomes increasingly complicated. PMID- 9330548 TI - Assessing health-related quality of life in HIV disease: key measurement issues. AB - The reliable and valid assessment of health-related quality of life (HRQoL) in human immunodeficiency virus (HIV) clinical research presents both familiar and unique challenges. Consistent with HRQoL research in general, measures of HRQoL in HIV disease must meet a set of standard psychometric properties, produce interpretable results, and be responsive to relatively small treatment effects. Furthermore, as clinical research for a range of diseases and conditions becomes increasingly global, HRQoL investigators are confronted with the formidable task of developing measures that are applicable across a range of cultures and languages within and across national boundaries. In this paper, we present a model of HRQoL to be applied across disease and conditions, with a discussion of the key measurement issues. We then briefly consider the natural history and treatment aspects of HIV that are relevant to HRQoL research. In the final sections of the paper, we describe the elements needed in an 'ideal' HRQoL/HIV instrument and propose a method for evaluating the degree to which current HRQoL measures address the challenges posed by HIV clinical research. PMID- 9330549 TI - Evidence for reliability, validity and usefulness of the Medical Outcomes Study HIV Health Survey (MOS-HIV). AB - The Medical Outcomes Study HIV Health Survey (MOS-HIV) is a brief, comprehensive measure of health-related quality of life (HRQoL) used extensively in human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS). The 35 item questionnaire includes ten dimensions (health perceptions, pain, physical, role, social and cognitive functioning, mental health, energy, health distress and quality of life (QoL) and takes approximately 5 minutes to complete. Subscales are scored on a 0-100 scale (a higher score indicates better health) and physical and mental health summary scores can be generated. The MOS-HIV has been shown to be internally consistent, correlate with concurrent measures of health, discriminate between distinct groups, predict future outcomes and be responsive to changes over time. Limited experience suggests acceptable reliability and validity in women, injecting drug users and African-American and lower socioeconomic status patients. The MOS-HIV is available in 14 languages and has been included as a secondary outcome measure in numerous clinical trials for all stages of disease. In several studies it has detected significant differences between treatments; in some cases concordant with conventional endpoints and, in others, discordant. The interpretation of scores is facilitated by an explanation in terms meaningful to the intended audience. Research is needed to compare the MOS-HIV to other strategies for HRQoL assessment in early HIV disease. PMID- 9330550 TI - Assessment of quality of life in early stage HIV-infected persons: data from the AIDS Time-oriented Health Outcome Study (ATHOS). AB - The development of new pharmaceutical interventions for persons with human immunodeficiency virus (HIV) infection has resulted in extended survival and a need for valid, reliable and responsive instruments to assess health-related QoL (HRQoL). This paper reviews the reliability and validity of an HRQoL instrument, the AIDS Health Assessment Questionnaire (AIDS-HAQ), among persons participating in an observational database of HIV infection. The AIDS-HAQ includes nine subscales: disability, energy, general health, pain, cognitive functioning, mental health, social functioning, health distress and symptoms. Individuals complete the AIDS-HAQ quarterly. Data are reported for 440 individuals entering the study with early HIV infection. Fifty-nine progressed to symptomatic disease and 109 to AIDS after 1 year. The subscales of the instrument resulted in high internal consistency reliability (range = 0.79-0.88). Concurrent validity data reflected the ability to distinguish between patients with increasing disease severity. In all domains, except cognitive functioning, individuals who progressed to AIDS had significant decrements (p < 0.01) in HRQoL compared with symptomatic and asymptomatic patients. Significant decrements (p < 0.01) were observed for disability, general health, energy and symptoms for patients who progressed to symptomatic disease from an asymptomatic status. Individuals who had decreasing CD4+ counts also had significant declines (p < 0.001) in disability, general health, social functioning, pain and symptoms. The AIDS-HAQ is an instrument that can be used when comparing group differences and within group changes in observational databases, naturalistic studies and clinical trials. PMID- 9330551 TI - The Quality of Well-Being scale in asymptomatic HIV-infected patients. HNRC Group. HIV Neural Behavioral Research Center. AB - We review applications of the Quality of Well-Being (QWB) scale for use in studies of human immunodeficiency virus (HIV)-infected patients. The QWB scale is a preference-weighted decision theory-based measure that summarizes outcomes in terms of quality-adjusted life years (QALYs). In order to validate the QWB scale for HIV-infected patients, the measure was administered in the University of California, San Diego (UCSD) HIV Neural Behavioral Research Center (HNRC). Data are presented for a cohort of 400 HIV-positive-infected men and 114 HIV uninfected male controls. The evidence suggests that the QWB scale is a significant prospective predictor of mortality. The QWB scale was concurrently associated with the number of CD4+ lymphocytes and ratings of neurological impairment based upon clinical evaluations. Further, the QWB scale was related to neuropsychological assessments derived from formal tests of cognitive functioning. Neuropsychological impairments may be associated with income loss for affected patients. The QWB scale scores were lower among patients with clinical depression. We conclude that the QWB scale is an appropriate general health outcome measure for use in observational studies and clinical trials for patients with HIV disease. PMID- 9330552 TI - Measuring quality of life in early HIV disease: the modular approach. AB - RATIONALE: to examine the reliability and validity of the General Health Self assessment, a modular questionnaire for self-assessment of quality of life (QoL) in human immunodeficiency virus (HIV) clinical trials and to describe the baseline QoL of participants in a large HIV clinical trial. DESIGN: the domains assessed include health perceptions, physical, psychological and role/social functioning, health care utilization and symptom distress. METHOD: 1,694 subjects with early HIV infection enrolled in the AIDS Clinical Trials Group Protocol 175 completed the scale at baseline. RESULTS: the domains demonstrated reliability, construct and discriminant validity. A worse QoL was associated with recent hospitalization and symptomatic status. Prior antiretroviral therapy was associated with higher health perceptions and well-being. The presence of symptom distress was related to lower QoL on the other scales. There was no relationship between QoL scales and the baseline CD4 count. Women showed a lower QoL than men on all scales, while ethnicity was related to differences in health perceptions and physical and psychological functioning. CONCLUSIONS: the General Health Self assessment shows excellent potential as a measure of QoL for HIV-infected patients in clinical trials. Further research is necessary to determine the responsiveness of the scale to clinical and immunological changes in HIV-infected individuals. PMID- 9330554 TI - Use of the MQoL-HIV with asymptomatic HIV-positive patients. AB - The purpose of this study was to determine the appropriateness of the recently developed Multidimensional Quality of Life Questionnaire for HIV/AIDS (MQoL-HIV) as a measure of quality of life (QoL) in cases of asymptomatic human immunodeficiency virus (HIV) infection. The MQoL-HIV is a 40-item instrument measuring ten domains (mental health, physical health, physical functioning, social functioning, social support, cognitive functioning, financial status, partner intimacy, sexual functioning and medical care) relevant to HIV infection. An overall QoL score, the MQoL-HIV Index, is a weighted composite of two domain scores. In a sample of 216 HIV-infected men and women, the MQoL-HIV distinguished acquired immune deficiency syndrome (AIDS), symptomatic and asymptomatic cases in overall QoL and in seven individual QoL domains. The index was responsive to perceived QoL changes over 5.5 months (r = 0.52). We also found the MQoL-HIV was less susceptible to ceiling effects in asymptomatic cases than was the Medical Outcomes Study (MOS) SF-20. These results suggest that the MQoL-HIV is a valid and reliable measure of QoL for both asymptomatic and symptomatic HIV infection. PMID- 9330553 TI - Applications of the Medical Outcomes Study health-related quality of life measures in HIV/AIDS. AB - The leading health status instruments in human immunodeficiency virus (HIV) research are based on the pool of items developed as part of the Medical Outcomes Study (MOS). The measures include the SF-20, MOS-HIV, SF-36, SF-12, SF-56, SF-38 (Patient Reported Status and Experience Survey (PARSE)), SF-21 and HIV Cost and Service Utilization Study (HCSUS) questionnaires. The instrument length ranges from 12 to 56 items, covering two to 11 dimensions. Completion requires from 2 to 14 minutes. Subscales are scored on a 0-100 scale (a higher score indicates better health); physical and mental health or overall summary scores are available for most of the measures. Three of the instruments are available in multiple languages. The instruments have been administered to over 20,000 persons with HIV in descriptive studies and clinical trials and there is substantial evidence for their reliability, construct and predictive validity and responsiveness. In several studies the measures have shown important differences between treatments. Although existing measures do not assess all domains relevant to HIV disease, additional subscales are available from the MOS pool. Some of the subscales may be prone to floor and ceiling effects. However, summary scales that encompass all of the subscales reduce this issue. Selection among MOS measures should be dictated by specific questions, the balance of available time and resources, and practical concerns. PMID- 9330555 TI - Performance of a new, HIV/AIDS-targeted quality of life (HAT-QoL) instrument in asymptomatic seropositive individuals. AB - OBJECTIVE: To evaluate the psychometric performance of a new human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS)-targeted quality of life (QoL) instrument in asymptomatic HIV-seropositive individuals. METHODS: 201 urban and rural, HIV-seropositive subjects were recruited to complete a 76-item pilot QoL measure developed using content provided in group process by seropositive individuals. Questionnaire responses from the full sample (n = 201) were used to identify dimensions and to reduce the number of items to 42. The responses to the retained items were then analysed for the asymptomatic subsample (n = 106). RESULTS: 9 multi-item dimensions were identified: overall function (OF), sexual function (SF), disclosure worries (DW), health worries (HW), financial worries (FW), HIV mastery (HM), life satisfaction (LS), medication concerns (MC) and provider trust (PT). The responses by asymptomatic subjects (74% male, 64% non-White and 63% homosexual/bisexual) revealed no substantial floor or ceiling effects, except for the PT dimension (where 44% were found to have scored the highest score). The internal consistency coefficients (Cronbach's alphas) were between 0.80 and 0.89 for six dimensions. The coefficients were lower for the SF (0.52), HM (0.67) and MC (0.48) dimensions. Construct validity assessments, using self-reported HIV disease-severity and sociodemographic variables, revealed some significant relationships (p < or = 0.05) for all dimensions except SF, MC and PT. CONCLUSIONS: The results suggested that five dimensions (OF, DW, HW, FW and LS) from the new instrument have good psychometric properties for asymptomatic HIV-seropositive individuals. These dimensions may be useful in the study of asymptomatic, seropositive individuals' QoL. Four dimensions (SF, HM, MC and PS) require additional refinement for this subpopulation. PMID- 9330556 TI - Psychometric validation of the revised Functional Assessment of Human Immunodeficiency Virus Infection (FAHI) quality of life instrument. AB - The revised Functional Assessment of Human Immunodeficiency Virus Infection (FAHI) quality of life (QoL) instrument has been updated and expanded to provide more complete and accurate coverage of human immune deficiency virus/acquired immune deficiency syndrome (HIV/AIDS)-related QoL. Factor analysis and the Rasch measurement model were used to determine a new subscale structure for the FAHI. The content of these subscales, including physical well-being (ten items, alpha = 0.91), function and global well-being (13 items, alpha = 0.86), emotional well being/living with HIV (10 items, alpha = 0.82), social well-being (eight items, alpha = 0.73), and cognitive functioning (three items; alpha = 0.75), reflect both general illness- and HIV/AIDS-specific QoL concerns: a total QoL score can also be calculated for the FAHI (44 items, alpha = 0.91). Psychometric evaluation revealed good internal consistency reliability for the FAHI and its subscales. In addition, construct validity, known groups validity and sensitivity to change were demonstrated by significant associations between the FAHI and additional indicators of functional status, psychological symptoms, stress and illness severity. In summary, the FAHI is a psychometrically sound instrument that captures multiple important dimensions of HIV/AIDS-related QoL. It is brief, easy to administer and score, has been translated into nine languages other than English and is appropriate for use in clinical trials and clinical practice. PMID- 9330557 TI - Measuring quality of life from the point of view of HIV-positive subjects: the HIV-QL31. AB - Assessment of the quality of life (QoL) of human immunodeficiency virus (HIV) infected subjects is often based on questionnaires in which the items or questions are not seen to be relevant by patients, nor by the users of the data obtained. It therefore seemed appropriate to return to the issue. The methodological and bibliographical research as well as the consultations we conducted convinced us that the elaboration of a new questionnaire was both necessary and possible. In order to do so, we adopted methodological principles based on the Sickness Impact Profile development methodology. First a bibliographical research was conducted in order to study instruments already used for HIV infection. Then, experts concerned with HIV infection and members of patients' associations were interviewed to assess how opportune the development of a new instrument could be. Following this, a methodology was established for the design and construction of the new instrument. One hundred and eighteen candidate questions were generated from an analysis of the content of 20 patients' interviews, which were subsequently submitted to 102 patients, to obtain finally a set of 31 questions from the interpretation of the results obtained from classic psychometric analysis and also from non-classic methods (item response theory and Rasch model). The concept being measured is the impact of illness being experienced by HIV-infected subjects from their own perspective. The range of health states covered by this questionnaire relates to fairly mild conditions. Rasch analysis of this set of 31 questions (HIV-QL31) shows that it corresponds to one unidimensional construct. A single score can be calculated by simple summation of dichotomous response options. This score is highly reliable (Cronbach's alpha coefficient = 0.93) and is also discriminant regarding the severity of clinical status. PMID- 9330558 TI - Health status assessment for the twenty-first century: item response theory, item banking and computer adaptive testing. AB - Health status assessment is frequently used to evaluate the combined impact of human immunodeficiency virus (HIV) disease and its treatment on functioning and well-being from the patient's perspective. No single health status measure can efficiently cover the range of problems in functioning and well-being experienced across HIV disease stages. Item response theory (IRT), item banking and computer adaptive testing (CAT) provide a solution to measuring health-related quality of life (HRQoL) across different stages of HIV disease. IRT allows us to examine the response characteristics of individual items and the relationship between responses to individual items and the responses to each other item in a domain. With information on the response characteristics of a large number of items covering a HRQoL domain (e.g. physical function, and psychological well-being), and information on the interrelationships between all pairs of these items and the total scale, we can construct more efficient scales. Item banks consist of large sets of questions representing various levels of a HRQoL domain that can be used to develop brief, efficient scales for measuring the domain. CAT is the application of IRT and item banks to the tailored assessment of HRQoL domains specific to individual patients. Given the results of IRT analyses and computer assisted test administration, more efficient and brief scales can be used to measure multiple domains of HRQoL for clinical trials and longitudinal observational studies. PMID- 9330559 TI - Summary and recommendations for future research. PMID- 9330560 TI - [Considerations on the fundamental structure and characteristics of the "Amae" phenomenon--clarification of the "Amae-theory (Takeo Doi)"]. AB - It is well-known that Takeo Doi tried to describe Japanese culture using the Japanese term "Amae". However, with many Japanese students of psychotherapy pointing out that his use of the term was arbitrary, his Amae-theory fell into confusion. Actually, with the single word Amae, he explained many heterogeneous psychological phenomena including pathological dependency as well as maternal separation. In this paper, I use my own clinical observations to clarify the Amae phenomenon and define it as follows: The "Aame" the Japanese usually experience in daily life differs from both pathological dependency (which M. Balint described as "ocnophilia"), and an affinity for friendly expanses in the therapeutic depressive position (described by M. Balint as "philobatism", by me as "sumu-akirameru"; c.f. Keiichi Nagayama: Considerations on the Guilt Feeling towards Mother and Maternal Separation using the Japanese Keywords "Sumu Akirameru" and "Sumanai", Psychiatria et Neurologica Japonica, 96: 83-108, 1994). In psychotherapy, Japanese patients only become able to form Amae connections with others after experiencing the two opposites (ocnophilia and philobatism) mentioned above. Although Amae is phenomenologically different from those extremes, it consists of two elements carrying some attributes from those extremes. One element consists of interpenetrating and mutually interdependent personal relations in a small familiar group; and the other element is a kind of protective and harmonious space in which the Japanese enjoy "temporary and partial regression in the service of the ego". As these two elements are both present in Amae, where dependence and independence are concerned, Amae toes the midline and has a double meaning. A fundamental principle of interpersonal relationships in Japanese society, Amae calls upon members of a small group to be moderately individualistic. If a member lacks ego flexibility regarding Amae and cannot obey this principle, he cannot adapt to a small familiar group. From the Western standpoint of individuality, Amae and interpenetrating personal relations in Japanese society must be regarded as regressive phenomena, whereas, for the Japanese, Amae is a personal skill necessary for social adaptation. Whereas Sumu Akirameru (Nagayama) and Philobatism (Balint) are ontological phenomena that tend to avoid external objects, Amae involves the need for relationships and is a somewhat socialized phenomenon in Japan. Although Amae and Sumu-Akirameru are different phenomena, they share several characteristics: 1) both tend towards protective harmonious spaces which have both specialty and boundaries; 2) things arise spontaneously and unintentionally in both phenomena; 3) the Japanese use both to confirm their sense of self; 4) both have the qualities of "corporality" and "living in". Because of these common areas, the Japanese tend to perceive Sumu-Akirameru and Amae as one experience, although they are different, separate phenomena. This tendency leads the Japanese to group behavior patterns, and unconsciously forces them into a double bind between individualization and group behavior patterns. This cultural tendency and phenomenological ambiguity of Amae itself allow the Japanese to easily project many kinds of psychological phenomena onto Amae. It is this projection deeply rooted in Japanese culture that confused the Amae-theory (Doi). Clarification of this cultural tendency not only contributes to the study of Japanese psychotherapy (Morita therapy, Naikan therapy), but also to that of "narcissism" and Preoedipal subjects in psychoanalysis. PMID- 9330561 TI - [The aggressivity in schizophrenics]. AB - Three young male patients with schizophrenia who developed aggressive acts towards themselves and others, suicide attempts, assaults, and a murder, are reported and discussed from the viewpoints of the language and the symbolization. The characteristics reported in proceeding researches were found in these three schizophrenics. It was when these patients interrogated, "What is the father?" that the psychosis was triggered and that the aggressivity closely connected with the conception of the death became manifest. These patients interrogated themselves as to being human and had to prove that they belonged to the human beings. Why the aggressivity in schizophrenics becomes manifest in this way? Because the symbolization didn't take place in schizophrenics and they were not subject to the internal death, necessary to be structured in the symbolic dimension. As a result, when they are asked, "What is the father? What is the human?" at the beginning of the psychosis, the manifests without symbolic articulation. The aggressivity in schizophrenics is considered as the function to murder the and to induce the internal death from the outside. It's Law-of the-Father that inhibits the manifestation of the for subjects in the symbolic dimension. However, Law-of-the-Father doesn't function in schizophrenics and returns in the delusion and the hallucination at the beginning of the psychosis when 'what is the father' comes in question. The absolute other exhibiting Law-of-the-Father orders the murder of the , and in consequence the aggressivity manifests by the orders. PMID- 9330562 TI - Debating the power of the placebo--experimental error or expectation mediated? PMID- 9330563 TI - Social support and long-term survivors of AIDS. AB - A naturalistic study design using ethnographic interviews was employed to elicit data about the phenomenon of being a long-term survivor of AIDS from 14 men and 6 women. Data were generated through multiple intensive open-ended interviews, demographic data sheets, and self-reported CD4 counts. Data were analyzed using latent and manifest content analysis techniques and the method of constant comparison. One of the dimensions that emerged from the data was "being in relation to others," the complex set of interpersonal relationships that have been renegotiated to maintain the reconstructed life. Specific ways of being in relation to others included dealing with one's family, renegotiating the friendship group, helping others with HIV, and developing a relationship with a higher power. The results of this study have implications for counseling people with HIV disease, and for nursing actions to enhance social support in this vulnerable group. PMID- 9330564 TI - The experiences of postmenopausal women with coronary artery disease. AB - The lifetime probability that a woman will develop coronary artery disease (CAD) is 46%. Hormonal changes, resulting in a shift in the ratio of the protective high density lipoproteins to low density lipoproteins dramatically increase the incidence of CAD in women after menopause. Historically, women have not been considered at risk for the development of CAD and clinical research has focused on men. The purpose of this descriptive study was to gain insight into the experiences of postmenopausal women after being diagnosed with CAD. Twelve postmenopausal participants were interviewed about their experiences since being diagnosed with CAD. The four major categories of data that emerged from this study were the effects of having a diagnosis of CAD, managing lifestyle changes resulting from this illness, identifying support systems, and adapting or coping with a diagnosis of CAD. PMID- 9330566 TI - Description of a self-care instrument for elders. AB - The psychometric properties of the self-care component of the Abilities Assessment Instrument were studied with elderly women. Thirty of the participants were cognitively impaired and 20 participants were not. Nonparametric statistics were used as the data were not normally distributed. Using the Spearman rank order correlation coefficient, test-retest reliability was rs = .79; p < .01 (n = 10) and interrater agreement was rs = .97; p < .01 (n = 9). Cronbach's alpha was .97. Between two content experts, the index of content validity was .89. Concurrent validity was assessed by comparing the self-care scores with the MMSE scores (rs = .91; p < .01). To establish construct validity, the self-care scores of those with dementia were shown to be statistically different from those who did not have dementia (Mann-Whitney U = 85). This study provided evidence for the reliability and validity of this self-care instrument. PMID- 9330565 TI - Parental experience and meaning construction during a pediatric health crisis. AB - This study explores parental lived experience following admission of their child to a pediatric intensive care unit. The interview data used were collected from 10 randomly chosen families from the Family Impact of Catastrophic Childhood Illness Project recruited during the early phase of critical care hospitalization of their child. A 3-stage contextual analysis procedure integrating interactional and contextual perspectives into Colaizz's phenomenological approach was used to reduce text data to thematic content. The analysis uncovered a multidimensional and holistic phenomenon consisting of four organizing concepts: initial boundary ambiguity, parents' coping patterns, family resources, and functioning of the family boundary. These results provide evidence of a collective family level perception of stress when experiencing the health crisis of a child and support further use of family stress perception as a family level phenomenon that represents family meaning construction during critical illness of a child. PMID- 9330567 TI - Health-promoting behaviors of black and white college women. AB - There is growing recognition that race and socioeconomic variables in health research demand greater attention. The investigators compared racial differences in health definition, health value, and health-promoting behavior of 62 pairs (N = 124) of Black and White college women matched on age, body mass index, and socioeconomic status. Both groups of women had similar definitions of health, valued health to the same extent, and reported similar levels of self actualization, health responsibility, exercise, and stress management. Black women, relative to White women, practiced fewer nutrition behaviors and had less interpersonal support. Interventions to reduce health risk associated with nutrition practices of Black women are warranted and further research is needed to explore the influence of the social structure of educational institutions on interpersonal relationships and other health behaviors. When socioeconomic status is taken into consideration, Black and White college women demonstrated more commonalities in health behavior than differences. PMID- 9330569 TI - [Sphingolipid biology in the central nervous system]. PMID- 9330568 TI - Using critical incident technique to inform aged and extended care nursing. AB - Flanagan's critical incident technique was used to explore the beliefs of a cross section of careers (both nursing and others) and consumers about the value of nursing and the nature of nursing's contribution in aged and extended care. The exploratory study found that nurses' being there, with and for residents, their families, and other health professionals, was influenced by two major themes that emerged from the data: personal and structural considerations. In this article, the potential of critical incident technique as a research method in nursing and for generating information about critical aspects of nursing work in aged and extended care is demonstrated. PMID- 9330570 TI - [Orphan nuclear receptor ROR alpha in cerebellum]. PMID- 9330571 TI - [Structure and function of Clostridium botulinum progenitor toxins]. PMID- 9330572 TI - [LIM domains: double zinc finger motifs involved in protein-protein interactions]. PMID- 9330574 TI - [Role of the steroidogenic acute regulatory (StAR) protein in the delivery of cholesterol to the inner mitochondrial membrane]. PMID- 9330573 TI - [Protein trafficking in the secretory and endocytic pathways]. PMID- 9330575 TI - [Tissue biosensor for determination of tetrodotoxin]. PMID- 9330576 TI - [Calorimetry of proteins (I)]. PMID- 9330577 TI - [Losartan (LORZAAR) and Losartan/hydrochlorothiazide (LORZAAR-PLUS)--their value in hypertension therapy. Symposium on the occasion of the 103rd Session of the German Society. Wiesbaden, 7 April 1997]. PMID- 9330578 TI - [Moxonidin in heart failure: new therapeutic principles suppress neurohumoral activation. Supplement to the Congress on "Heart Failure" of the working group on cardiac insufficiency at the European Society for Cardiology. Cologne, May 1997]. PMID- 9330579 TI - [Irbesartan--a new AT1-receptor antagonist (AT1-blocker). Angiotensin-II-receptor antagonists: perspectives of a new therapeutic principle, Wiesbaden, 6 April 1997]. PMID- 9330581 TI - Receptor regulation--food for thought. PMID- 9330582 TI - The role of glucagon-like peptide 1 in the regulation of glucose homeostasis and satiety. PMID- 9330580 TI - Glucocorticoid osteoporosis--mechanisms and management. AB - Glucocorticoids are potent osteopenic agents, producing negative calcium and bone balance via actions at many sites. The most significant adverse effects of glucocorticoid drugs on the skeleton are probably a direct inhibition of matrix synthesis by the osteoblast, reductions in calcium absorption in both the gut and the renal tubule, and the production of hypogonadism, particularly in men. Reductions in bone density of 10-40% result, the loss being more marked in trabecular bone and in patients receiving a high cumulative dose of the steroid. Fractures occur in about 30% of individuals who take these drugs for an average of 5 years. Bone loss is reversible when glucocorticoid treatment is withdrawn. Bone density can also be increased by sex hormone replacement in those with demonstrable deficiency, by bisphosphonates, and possibly by vitamin D metabolites. All patients treated with glucocorticoids for more than 6 months should be considered for bone densitometry and be offered appropriate drug treatment if values are towards the lower end of the young normal range or if there is already evidence of fractures occurring after minimal trauma. With this approach, the significant morbidity associated with steroid osteoporosis might be substantially avoided. PMID- 9330583 TI - A long-sought needle in the haystack: the multiple endocrine neoplasia type 1 gene. PMID- 9330584 TI - Novel pharmacological approaches to the prevention and treatment of non-insulin dependent diabetes mellitus. PMID- 9330585 TI - Effects of long-term total fasting and insulin on ob gene expression in obese patients. AB - In the present study the effect of long-term fasting (6 days) on obese (ob) gene expression was examined in nine severely obese females of 34 +/- 3 years and with a body mass index of 46.4 +/- 2.3 kg/m2. Six days of fasting induced a significant weight loss (126.8 +/- 5.3 vs 120.5 +/- 5.1 kg, P < 0.0001). Insulin stimulated glucose uptake (hyperinsulinemic, euglycemic clamp, insulin infusion rate 1.5 mU/kg per min) was markedly reduced following fasting (M-value 5.96 +/- 0.74 vs 2.79 +/- 0.23 mg/kg per min, P < 0.0001). Ob mRNA/beta-actin concentration in fat biopsies from abdominal subcutaneous adipose tissue was unchanged after 6 days of fasting (1.50 +/- 0.40 vs 1.47 +/- 0.36 arbitrary units, not significant), whereas serum leptin levels decreased significantly from 53.8 +/- 4.7 to 30.7 +/- 2.0 ng/ml (P < 0.0001) during the same period. No significant correlations were found between insulin-stimulated glucose uptake and serum leptin concentration, either prior to the fast or after the fast. Serum leptin levels were unchanged by hyperinsulinemia for 3 h during the clamp prior to the fast, while hyperinsulinemia for 3 h after 6 days of fasting increased serum leptin by 25% (P < 0.01). In conclusion, 6 days of fasting reduced serum leptin by about 40%. In contrast, ob mRNA in abdominal subcutaneous adipose tissue was unchanged. Furthermore, after 6 days of fasting insulin was able to increase the serum level of leptin significantly, indicating that the effect of insulin on the level of leptin is dependent on the nutritional state. PMID- 9330587 TI - Increased fracture frequency in adult patients with hypopituitarism and GH deficiency. AB - Fracture frequency was studied in 107 hypopituitary patients with GH deficiency (GHD) (69 men, mean age 53 years, range 18-74 and 38 women, mean age 54 years, range 31-73). Routine hormonal replacement therapy was given, except GH. Five male patients and 15 female patients with untreated hypogonadism were allocated to a separate group. The mean duration of hypopituitarism was 13.4 years. The prevalence of a history of fractures was assessed using questionnaires. A subsample of the Goteborg WHO MONICA Project was used as a reference population (n = 323). The total fracture frequency was threefold higher (P < 0.001) in patients (24.1%) compared with controls (8.7%) (odds ratio 3.49) (1.85-6.56; 95% confidence intervals). In men (n = 64) the fracture frequency was 25.0%, compared with 7.8% among the controls (P < 0.001). In women (n = 23) the fracture frequency was 21.7%, compared with 9.5% among the controls (P = 0.08). The odds ratios for fracture frequency were 3.97 (1.81-8.40; 95% confidence intervals) and 2.64 (0.89-7.81; 95% confidence intervals) in men and women respectively. In conclusion, adult hypopituitary patients with GHD had a threefold increased fracture frequency compared with controls. Further studies are needed to ascertain whether long-term recombinant human GH treatment can reduce the fracture rate in hypopituitary patients with GHD. PMID- 9330586 TI - Vitamin E and nicotinamide have similar effects in maintaining residual beta cell function in recent onset insulin-dependent diabetes (the IMDIAB IV study) AB - OBJECTIVE: Protection of residual beta cell function at the time of diagnosis of insulin-dependent diabetes mellitus (IDDM) by intensive insulin therapy and the addition of nicotinamide (NA) has been established. The objective of this study was to evaluate the effect of a free oxygen radical scavenger such as vitamin E (Vit E) on residual beta cell function and parameters of metabolic control in patients with recent onset IDDM undergoing intensive insulin therapy. DESIGN: The effect of Vit E was compared with that of NA (control group) in a randomized multicentre trial. METHODS: Eighty-four IDDM patients between 5 and 35 years of age (mean age 15.8 +/- 8.4 (s.d.) years) entered a one year prospective study. One group of patients (n = 42) was treated with Vit E (15 mg/kg body weight/day) for one year; the other group (n = 42) received NA for one year (25 mg/kg body weight/day). All patients were under intensive insulin therapy with three to four injections a day. Basal and stimulated (1 mg i.v. glucagon) C-peptide secretion, glycosylated haemoglobin and insulin dose were evaluated at diagnosis and at three-monthly intervals up to one year. RESULTS: Preservation and slight increase of C-peptide levels at one year compared with diagnosis were obtained in the two treated patient groups. No statistically significant differences were observed in basal or stimulated C-peptide levels between the two groups of patients for up to one year after diagnosis. Glycosylated haemoglobin and insulin dose were also similar between the two groups; however patients receiving Vit E under the age of 15 years required significantly more insulin than NA-treated patients one year after diagnosis (P < 0.04). CONCLUSIONS: Our data indicate that Vit E and NA possess similar effects in protecting residual beta cell function in patients with recent onset IDDM. Since their putative mechanism of protection on beta cell cytotoxicity is different, combination of these two vitamins may be envisaged for future trials of intervention at IDDM onset. PMID- 9330588 TI - Circulating non-22 kDa growth hormone isoforms in healthy children of normal stature: relation to height, body mass and pubertal development. AB - The proportion of non-22 kDa GH isoforms was evaluated in 93 healthy children (48 boys aged 6.8-18.4 years and 45 girls aged 3.9-18.4 years) of normal stature (height +/- 2 s.d. score) at different stages of puberty. In addition, correlations among the proportion of non-22 kDa GH isoforms, auxology, spontaneous GH secretion and biochemical measurements were investigated. Serum non-22 kDa GH levels, expressed as percentage of total GH concentration in the samples, were determined by the 22 kDa GH exclusion assay, in which monomeric and dimeric 22 kDa GH are removed from serum and the non-22 kDa GH isoforms are quantitated using a polyclonal antibody GH assay. Samples were selected from spontaneous GH peaks in 24-h GH profiles. For boys, the median proportion of non 22 kDa GH isoforms was 8.5% (range 3.2-26.6%) and for girls it was 9.6% (1.8 17.4%), with no influence of age and no sex-related difference in prepubertal (boys, 7.2%; girls, 8.8%) or pubertal children (boys, 9.1%; girls, 9.9%). However, the median proportion of non-22 kDa GH isoforms was significantly higher in pubertal boys (9.1%) than in prepubertal boys (7.2%; P = 0.03). In pubertal boys, height S.D. scores (SDS) were inversely correlated to the proportion of non 22 kDa GH isoforms (r = -0.38; P = 0.02), especially at mid-puberty (r = -0.7; P = 0.01), indicating that the presence of increased amounts of circulating non-22 kDa GH isoforms was associated with less growth. In prepubertal children, positive correlations between non-22 kDa GH and weight SDS (r = 0.46; P = 0.03), weight-for-height SDS (r = 0.51; P = 0.01) and body mass index (r = 0.42; P = 0.04) were observed. No significant correlations were seen with spontaneous GH secretion or measurements of IGF-1, IGF-binding protein-3, insulin and leptin. These findings in normal children indicate that the proportion of circulating non 22 kDa GH isoforms may have physiologic significance for growth and metabolism in different stages of development, and emphasize the importance of evaluating the circulating ratio of 22 kDa and non-22 kDa GH in children with growth disorders. PMID- 9330589 TI - Evaluation and treatment of persistent thyroglobulinemia in patients with well differentiated thyroid cancer. AB - Whereas in the past a negative diagnostic 131I whole body scan (WBS) was interpreted as the lack of significant residual or recurrent thyroid cancer, today the patient with negative WBS and measurable serum thyroglobulin (Tg) presents a diagnostic and therapeutic dilemma. Previous studies have shown a high rate of visualization of uptake and a decrease in Tg after one or more therapeutic doses of 131I. In order to further assess the significance of this finding, retrospective analysis of patients with persistent thyroglobulinemia and negative WBS was performed for evidence of surgically amenable disease. Seven out of seventeen patients had neck ultrasound and/or computerized tomography (neck +/ chest) showing the presence of pathologically confirmed malignant masses ranging from 1 to 4 cm in size. Their serum Tg while on L-thyroxine ranged between 2.4 and 1173 pmol/l. Removal of the identified masses resulted in a greater than 75% reduction in serum Tg in four out of five patients in the group. One patient achieved a serum Tg of < 1.5 pmol/l while hypothyroid. Empiric 131I treatment of eleven patients with persistent thyroglobulinemia resulted in demonstrated uptake on post-therapy scan in seven. Further study is needed to compare the efficacy, safety and cost of a diagnostic approach to radiologically identify and surgically resect identified disease versus empiric therapeutic 131I treatment and high-dose WBS in this group of patients. Patients with negative WBS and persistent thyroglobulinemia, even to levels < 4.5 pmol/l, may have significant foci of thyroid cancer in surgically accessible areas. This suggests the need for a redefinition or clarification of the term 'recurrence' in thyroid cancer. PMID- 9330590 TI - Absence of angiotensin II type 1 receptor gene mutations in human adrenal tumors. AB - Regulatory actions of angiotensin II (AngII), which is involved in the pathophysiology of hypertension and also participates in cell proliferation and cell differentiation, are mainly mediated by AngII type 1 (AT1) receptor. Recently, activating mutations of receptors causing hyperfunctioning endocrine diseases have been described in the case of the TSH and LH receptors, implicating that such mutations might occur in other G-protein-coupled receptors. Furthermore it seems to be possible that genetic variations of AT1 receptor have an influence upon the action of AngII. Therefore, we searched by sequence analysis of the coding region of AT1 receptor gene for activating mutations and genetic polymorphisms in 56 human adrenal tumors (16 aldosterone-producing adenomas, 10 cortisol-producing adenomas, 1 aldosterone-producing carcinoma, and 29 incidentalomas). We were not able to identify any activating mutation in the coding region of AT1 receptor gene. We conclude that activating mutations of the AT1 receptor are not a major cause of the development of adrenal adenomas, if at all. In addition, polymorphic subtypes of AT1 receptor do not seem to play a major role in the pathogenesis of these tumors, even though a tendency towards a higher frequency of the polymorphic base substitution at position 573 (T573-->C) in cortisol-producing tumors needs to be further evaluated. PMID- 9330591 TI - An unusual case of papillary carcinoma of the thyroid with cutaneous and breast metastases only. AB - Cutaneous metastases of thyroid carcinoma are infrequent and, when present, are usually located in the vicinity of a widespread primary tumor. Breast metastases from these tumors are even less common. We report the case of a 64-year-old female with a toxic multinodular goiter in whom a fine-needle biopsy, performed in 1985 at the age of 52, was suggestive of papillary carcinoma of the thyroid. Total thyroidectomy for a papillary carcinoma, follicular variant, was performed in 1988. Four months after surgery, a cutaneous metastasis was discovered in the right thigh. Surgical excision of the lesion followed by treatment with radioactive iodine decreased serum Tg levels from 7495 to 3.3 micrograms/l. Under suppressive therapy with L-thyroxine, serum Tg remained undetectable for the next 4 years. Then, serum Tg levels rose to 3.9-5.6 micrograms/l and a second cutaneous metastasis was removed from the abdominal wall. The patient was again treated with radioactive iodine and the post-treatment whole-body scan did not show any area of increased uptake of the radionuclide. However, serum Tg levels under suppression with L-thyroxine remained elevated at 4-20 micrograms/l for the next 2 years. In August 1995, a 1.5 cm nodule was found in the right breast. Cytological examination was suggestive of a breast metastasis from thyroid carcinoma and the lesion was removed by enucleation. This proved to be a metastasis from a papillary carcinoma of the thyroid. Elevated (19-44 micrograms/l) serum Tg levels persisted postoperatively. A third cutaneous metastasis was revealed by 131I scintigraphy in the right buttock and surgically removed in December 1996. Serum Tg levels have remained undetectable since then. To the best of our knowledge, this is a unique case of a papillary carcinoma of the thyroid with a propensity to metastasize only to the skin and breast during a follow-up of 11 years. PMID- 9330592 TI - Starvation-induced increase in the parathyroid hormone/PTH-related protein receptor mRNA of bone and kidney in sham-operated and thyroparathyroidectomized rats. AB - Parathyroid hormone (PTH) acts on bone and kidneys by binding to PTH/PTH-related protein (PTHrP) receptors and regulating calcium (Ca) and phosphorus (P) homeostasis. PTH/PTHrP receptor mRNA was expressed at high levels in PTH target tissues such as the kidneys and bone including the calvaria, femur, and tibia. Because short-term starvation influences Ca and P ion homeostasis, we measured changes in PTH/PTHrP receptor mRNA expression in the bone and kidneys. Food deprivation for 3 days decreased the serum Ca and P concentrations, and reinstitution of feeding for 2 days normalized the serum Ca level and significantly increased the serum P level. Concomitantly, rat immunoreactive PTH (riPTH) was increased during starvation and returned to the control level after 2 days of subsequent feeding. Serum 1 alpha, 25-dihydroxyvitamin D3 (1,25(OH)2D3) concentrations did not significantly change during starvation and subsequent feeding. Starvation up-regulated PTH/PTHrP receptor mRNA expression in both bone and kidney. The effects of food deprivation on the receptor transcript abundance were greater in bone (threefold increase compared with control) than in the kidney (1.8-fold increase), whereas the mRNA level increase by food deprivation was more rapid in the kidneys than in bone. The PTH-induced adenylyl cyclase activity of renal membranes increased in starvation. Feeding after starvation normalized the mRNA levels in both tissues. Serum PTH depression, initiated by thyroparathyroidectomy, did not affect PTH/PTHrP receptor mRNA levels in bone and kidney in rats that were fed or starved for 3 days. The abundance of receptor mRNA in bone and kidney was significantly lower in fed rats given either corticosterone or vehicle than in starved rats. These data indicate that starvation induces PTH/PTHrP receptor mRNA expression in bone and kidney, independently of serum PTH and corticosterone concentrations. The factors leading to up-regulated receptor mRNA induced by starvation remain unknown. PMID- 9330593 TI - The steroid antagonist RU486 given at pro-oestrus induces hypersecretion of follicle-stimulating hormone from oestrus afternoon to early metoestrus in the rat. AB - Administration of the steroid antagonist RU486 to cyclic rats at pro-oestrus blunts the preovulatory surge of LH and suppresses the first and second surges of FSH. In addition, administration of oestradiol to RU486-treated rats reactivates the LH surge the following day. The present study explored the effects of RU486 (4 mg/0.2 ml oil), administered at 0800 h on the day of pro-oestrus, on serum FSH and LH concentrations through oestrus and early metoestrus. RU486 induced a hypersecretion of FSH, which started at 1400 h on the day of oestrus and was maintained until 0800 h on the day of metoestrus. Because the timing and magnitude of this secretion of FSH were similar to those of the periovulatory secretion of FSH during pro-oestrus and early oestrus in intact cyclic rats, we investigated the effects of: 1) LHRH antagonist (LHRHa) injected at either 0900 h or 2000 h on the day of oestrus, 2) oestradiol benzoate injected at 1600 h on the day of pro-oestrus and at 0900 h on the day of oestrus, 3) bovine follicular fluid (bFF) given either at 1100 h or at 2000 h on the day of oestrus, or 4) adrenalectomy (ADX) at 1100 h on the day of oestrus, on serum FSH and LH concentrations at 1800 h on the day of oestrus and at 0200 h on the day of metoestrus in rats injected with RU486 at pro-oestrus. The results showed that 1) both components (late oestrus and early metoestrus) of FSH hypersecretion in RU486-injected rats in pro-oestrus were inhibited by oestradiol benzoate and bFF, 2) the metoestrous component was not affected by LHRHa, whereas the oestrous component was partially reduced, and 3) ADX partially reduced serum FSH concentrations only on the day of metoestrus, possibly because, as the serum concentrations of corticosterone reflected, the antiglucocorticoid activity of 4 mg RU486 lasted only 24 h. The results support the hypothesis that blockade of progesterone actions at pro-oestrus results in the maintenance of the daily neural signal that activates the release of gonadotrophins. Whereas the expression of LH secretion requires high levels of oestradiol, FSH secretion is expressed against a background of low oestradiol levels. The results of this study also indicate that the release of FSH during oestrus and metoestrus in rats injected with RU486 at pro-oestrus is a consequence of the lack of ovarian negative feedback inhibition on the pituitary. PMID- 9330594 TI - Effects of castration on early postnatal development of male accessory sex glands in the domestic pig. AB - In the neonatal pig there is a remarkable production of steroids by the testes for the first few weeks after birth. Several androgens and estrogens reach a peak at about one month of age. In order to gain an understanding of the significance of this early steroid secretion we examined the effect on accessory sex glands of removal of the testes before the peak in these compounds would have occurred. Pigs were castrated (n = 38) at 2-3 weeks of age, with littermates serving as intact controls (n = 33). Animals were killed at ages ranging from 4-12 weeks. Blood samples were taken and both bulbourethral (BU) and vesicular glands (VG) were removed, as well as the testes of intact males. Organ pairs were weighted and samples fixed for histological examination. Plasma samples were stored at -20 degrees C until assayed, without extraction, for testosterone, dehydroepiandrosterone sulfate (DHEAS) and estrone sulfate (E1S) by radioimmunoassay. Of the hormones measured, plasma DHEAS concentrations were highest, but variable over the time period (304.2 and 75.6 nmol/l; 87.7 and 21.8 ng/ml at 5 and 12 weeks respectively). E1S declined steadily from 76.6 to 5.8 nmol/l (20.7 to 1.56 ng/ml). Testosterone levels were lowest but rose from 2.67 to 9.54 nmol/l (0.77 to 2.75 ng/ml). No steroids were clearly detectable in samples from castrated males. Testes weights (wt) increased fourfold, as did body wt for both intact and castrate males. Both BU and VG showed absolute increase in wt (3.5x and 5x respectively) in intact males, and each was about 2.8x greater than in castrates (mg/kg body wt). Histological sections were markedly distinctive for both BU and VG between intact and castrate animals, and a lack of developmental changes in both glands was noted in the castrates. Our findings provide clear evidence of an influence of the testes on accessory sex glands in the early postnatal life of the pig. PMID- 9330595 TI - Interactions between interleukin-1 beta, nitric oxide and prostaglandin E2 in the rat ovary: effects on steroidogenesis. AB - It has been reported that interleukin (IL)-1 beta induces the synthesis of both nitric oxide (NO) and prostaglandin (PG)E2 in cultures of dispersed ovarian cells and exerts cytotoxic effects on these cells. Since PGE2, NO and IL-1 beta have been implicated as modulators of steroidogenesis, experiments have been undertaken to determine how IL-1 beta-induced NO and PGE2 production may affect steroidogenesis in cultures of ovarian dispersates obtained from untreated adult oestrous rats and to compare the action of IL-1 beta in cultures of granulosa/luteal (GL) cell-only cultures and GL cells co-cultured with peritoneal macrophages. IL-1 beta significantly increased the production of NO (assessed by nitrite measured in the culture medium) and PGE2 in cultures of ovarian dispersates but had no effect on cultures of GL cells in which NO production was typically very low and PGE2 production was undetectable. In contrast both NO and PGE2 were high in co-cultures and were not significantly altered by the addition of IL-1 beta. The NO donor sodium nitroprusside inhibited steroidogenesis in cultures of ovarian cells in a dose-dependent manner while PGE2 had a stimulatory effect. Concomitant inhibition of NO production with aminoguanidine and PG production with indomethacin resulted in a significant enhancement of basal progesterone production in these cultures. Finally, IL-1 beta inhibited progesterone responses to forskolin and PGE2 in ovarian dispersates: an effect not observed in GL cell-only cultures nor in co-cultures in which forskolin induced progesterone production is always inhibited. No cytotoxic effects of IL-1 beta during the 48 h period of culture were observed. A comparison of the steroidogenic NO and PGE2 responses to IL-1 beta in the three culture models suggests that (i) the response to IL-1 beta observed in ovarian dispersates could be due to cytokine activation of resident and infiltrating macrophages or that the action of IL-1 beta requires some other heterologous cell-cell contact, and (ii) IL-1 beta acts indirectly on signal transduction pathways which stimulate steroidogenesis. PMID- 9330596 TI - Role of the testis in the response of the pituitary-testicular axis to nitric oxide-related agents. AB - Nitric oxide (NO) is generated from the guanidine group of L-arginine by NO synthases (NOS) in a wide variety of tissues, including endocrine organs. In order to discriminate between central and local effects of NO-related agents on the pituitary-testicular axis, adult rats were injected intraperitoneally with 1 g/kg body weight (bw) L-arginine methyl ester (L-AME, an exogenous substrate of NOS), 0.5 mg/kg bw sodium nitroprusside (SNP, an NO donor) or vehicle (0.9% NaCl) or intratesticularly with 2 mg/testis L-AME, 2 micrograms/testis SNP or 25 microliters vehicle, and killed at 60 or 120 min after treatment. Both intraperitoneal and intratesticular administration of L-AME had the same effects: a decrease in the serum concentrations of LH and testosterone and in those of testosterone in the testicular interstitial fluid. However, treatment with SNP was more effective when given intratesticularly, inducing a decrease in serum and interstitial fluid testosterone concentrations, without significant changes in LH concentrations. Furthermore, when rats were injected intraperitoneally with 4 mg L-AME (the same dose as that given intratesticularly), serum LH concentrations were not changed. In addition, L-AME administration was not effective in modifying serum LH concentrations in castrated rats. To test the possible role of Leydig cells, the effects of systemic administration of L-AME were studied in rats depleted of Leydig cells by treatment with ethylene dimethane sulphonate. In these animals L-AME significantly decreased serum LH concentrations. To study the role of macrophages in this system, rats depleted of testicular macrophages by the liposome-suicide approach were injected intraperitoneally (1 g/kg bw) or intratesticularly (2 mg/testis) with L-AME or vehicle, 10 days after macrophage depletion, and killed at 120 min after treatment. The effects of L-AME on serum LH concentrations were blocked when the drug was administered intratesticularly. PMID- 9330597 TI - Insulin-like growth factor-I stimulates myofibrillar genes and modulates atrial natriuretic factor mRNA in rat heart. AB - We have investigated the effect of a 6-day infusion of recombinant human (rh) IGF I (0.3-1.0 mg/day) or rhGH (200 mU/day) into normal and hypophysectomized rats on the ventricular expression of myofibrillar genes (alpha- and beta-myosin heavy chain (MHC), skeletal and cardiac alpha-actin) and of atrial natriuretic factor (ANF). In normal rats, beta-MHC was not detectable either before or after IGF-I or GH, but alpha-MHC mRNA increased significantly (twofold) with GH (not statistically significant for IGF-I). In contrast to normal rats, hypophysectomized rats did not express alpha-MHC either before or after IGF-I or GH, but beta-MHC was strongly expressed and significantly stimulated (1.8-fold) by IGF-I (not statistically significantly with GH). Skeletal alpha-actin expression remained unchanged during IGF-I or GH treatment of normal rats, but was enhanced by both IGF-I and GH (2.5- and 2.8-fold respectively) in hypophysectomized rats. Expression of cardiac alpha-actin in normal and hypophysectomized rats was not altered by either treatment. IGF-I and GH decreased ventricular expression of ANF in normal rats by 63% and 45% respectively, but did not influence ANF expression in hypophysectomized rats. Our results show that IGF-I and GH (possibly via IGF-I) stimulate expression of myofibrillar genes and modulate ANF mRNA concentrations in rat heart ventricles in vivo, depending on the hormonal status of the animals. However, neither IGF-I nor GH caused a shift from the beta- to the alpha-MHC isoform in hypophysectomized rats. PMID- 9330598 TI - The colorectal tumor suppressor APC and its partners. PMID- 9330599 TI - Infrequent mutation of the H-cadherin gene on chromosome 16q24 in human breast cancers. AB - To investigate the molecular basis of altered expression of the H-cadherin gene, we used polymerase chain reaction-single strand conformation polymorphism and DNA sequencing to examine the H-cadherin gene in 48 primary breast cancers in which loss of the long arm of chromosome 16 had been detected. We identified no mutations other than somatic 5-bp deletion within the coding region in a single tumor. The very low frequency of mutation found in these experiments suggests that H-cadherin is usually not a primary target for carcinogenesis in human breast cancers, and that reduction of its expression is likely to be a consequence of some other genetic event(s). PMID- 9330600 TI - Inhibitory effects of diallyl disulfide or aspirin on 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine-induced mammary carcinogenesis in rats. AB - Modifying effects of diallyl disulfide (DAD), aspirin or DL-alpha difluoromethylornithine (DFMO) on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced mammary carcinogenesis in SD rats were investigated. A total of 166 female rats, 6 weeks old, were divided into 8 groups. They were fed a high fat diet throughout the experiment. Starting at 7 weeks of age, groups 1-4 were given PhIP (85 mg/kg body weight in corn oil) by gavage 8 times in 10 days, and groups 5-8 were given corn oil alone. For the beginning 4 weeks, groups 2 and 5 were given DAD at 200 ppm in diet. Similarly groups 3 and 6, and groups 4 and 7 were given aspirin (400 ppm) and DFMO (400 ppm), respectively. Mammary carcinomas were only recognized in groups 1-4 at the termination (25 weeks after the start of experiment). Multiplicity (mean number/rat) of neoplasms in group 2 (PhIP+DAD, 0.90/rat) and group 3 (PhIP+aspirin, 1.37/rat) was significantly smaller than that in group 1 (PhIP alone, 2.45/ rat) (P < 0.005 and P < 0.05, respectively). These results indicate that dietary intake of DAD or aspirin during the time corresponding to initiation phase has chemopreventive potential on PhIP-induced mammary carcinogenesis in rats. PMID- 9330601 TI - Retroviral introduction of the p16 gene into murine cell lines to elicit marked antiproliferative effects. AB - The p16 gene is a candidate tumor suppressor, because mutation of the gene has been reported in many transformed cell lines and some primary tumor tissues. We have examined this possibility in murine cell lines (NIH3T3 and RSV-M) which lack p16 gene expression. Full-length human p16 cDNA was obtained from a HeLa cell line using polymerase chain reaction amplification. We constructed two separate retrovirus vectors carrying this p16 cDNA. First, we transduced the p16 cDNA into the murine cell lines using a retrovirus vector harboring the neomycin-resistance gene. The p16 gene-transduced cells formed no colonies after selection with G418, in contrast to the vector-transduced cells. Next, we used another retrovirus vector that expresses both the p16 cDNA and the Lac Z gene, which enabled us to distinguish affected cells from unaffected ones. Proliferation of the p16 gene transduced cells was markedly inhibited and morphological change in the cells was also observed. Thus, we concluded that the p16 gene has an antiproliferative effect on the cell cycle and that the loss of its function may play a major role in dysregulated proliferation of the cells. PMID- 9330602 TI - Analyses of the APC and TGF-beta type II receptor genes, and microsatellite instability in mucosal colorectal carcinomas. AB - APC and transforming growth factor-beta type II receptor (TGF-beta RII) gene mutations, and microsatellite instability have been found in sporadic colorectal carcinomas. To clarify further the early alterations in colorectal carcinogenesis, we investigated these genetic changes in 23 protruding- and 24 superficial-type mucosal colorectal carcinomas. TGF-beta RII gene mutations and microsatellite instability were rarely found in these lesions. Nevertheless, APC was mutated in 16 of the 47 (34.0%) mucosal colorectal carcinomas and was significantly more frequently mutated in protruding- (I) and superficial elevated type (IIa) (14/32, 43.8%) than in other superficial-type (IIa+IIc, IIb, IIc, and IIc+IIa) (2/ 15, 13.3%) mucosal colorectal carcinomas (P < 0.04). These results indicate that the APC gene may be involved from the beginning in the tumorigenesis of many early colorectal carcinomas, particularly of the protruding and superficial elevated types. However, there might be a distinct pathway for other superficial-type colorectal carcinomas, possibly not involving APC as an initial step of tumorigenesis. PMID- 9330605 TI - P-glycoprotein is positively correlated with p53 protein accumulation in human colorectal cancers. AB - To explore the relationship between mutant p53 and Pgp expression, we have examined the levels of both proteins in human colorectal adenocarcinomas. Serial frozen sections of 40 surgical samples were stained with an anti-Pgp (MRK16) and two different anti-p53 protein antibodies (Abs), PAb421 and PAb1801. Nineteen (47.5%) of 40 samples examined were positive for Pgp, and 18 (45%) of 40 were positive for p53. The samples that stained positively with PAb421 also stained positively with PAb1801. Pgp expression was detected in 13 (76.5%) of 17 samples that were positive for p53 using PAb421 and in 15 (83.3%) of 18 samples that were positive for p53 using PAb1801. Thus, we found that p53 and Pgp were co-expressed in a significant number of samples (P < 0.002). There was no relationship between Pgp or p53 protein accumulation and histologic grade or stage. The present results demonstrate that Pgp expression is closely associated with p53 protein accumulation in human colorectal cancers. These data provide evidence to support the idea that mutant p53 activates the MDR1 gene in vivo. PMID- 9330603 TI - Isolation of a novel gene showing reduced expression in metastatic colorectal carcinoma cell lines and carcinomas. AB - To investigate genes involved in metastatic stages of cancer, we analyzed expression of mRNAs in three cell lines derived from murine colon adenocarcinoma 26 by means of a differential display method. Each of these lines exhibits distinct metastatic characteristics. Among many bands representing different expression patterns in the display, we confirmed by northern analysis that a gene corresponding to one amplified fragment, termed grm2 (gene related to metastasis 2), was expressed more abundantly in NL4, the derivative with the lowest metastatic potential, than in cell lines NL17, an experimentally metastatic derivative, and in NL22, a spontaneously metastatic derivative. Using the grm2 fragment as a probe, we isolated murine cDNA clones and subsequently human cDNA clones corresponding to the GRM2 gene. The human and mouse homologues both encode proteins of 600 amino-acid residues, which show weak homologies to proteins belonging to the myosin family. When we examined the expression levels of this novel gene in human colon cancers and in corresponding metastatic foci, we found that in more than half of these tissues, expression was significantly reduced in association with malignant potential. Our results imply that in humans the GRM2 gene product may regulate the metastatic phenotype of some colorectal cancers. PMID- 9330604 TI - Telomerase activity and metastasis: expansion of cells having higher telomerase activity within culture lines and tumor tissues. AB - Tumor cells with metastatic potential may have a high telomerase activity that augments telomeric DNA repeats, allowing the cells to escape from the inhibition of cell proliferation due to shortened telomeres. We examined the expression level of telomerase activity using the telomeric repeat amplification protocol among a series of cell lines obtained by repeated transplantation of a mouse fibrosarcoma. The lines could be grouped into three; one has no metastatic potential, and the other two show metastatic abilities after intravenous or subcutaneous injection. Comparison of their telomerase activity indicated that more malignant lines had higher activity. A similar relation was seen in metastatic nodules formed through clonal expansion from the heterogeneous population of inoculated cells; clonality was monitored in terms of variable patterns of subtelomeric repeats. The results suggest that a high level of telomerase activity may not be requisite for metastasis, but may confer a propensity to dominate in a tumor tissue. PMID- 9330606 TI - The induction of cytotoxic T lymphocytes against HLA-A locus-matched lung adenocarcinoma in patients with non-small cell lung cancer. AB - To induce cytotoxic T lymphocytes (CTL) against non-small cell lung cancer (NSCLC) efficiently, the induction of CTL was attempted using HLA-A locus-shared allogeneic NSCLC cells. T cells derived from either tumor tissue specimens or the regional lymph nodes of patients with NSCLC were stimulated twice or three times with an HLA-A2/A24-positive NSCLC cell line (PC-9), and thereafter the cytotoxic activity was examined by 51Cr-release assay. In patients with HLA-A24/ adenocarcinoma, anti-PC-9 cytotoxicity was induced in all 6 patients tested. Anti PC-9 cytotoxicity was induced in 2 out of 5 patients with HLA-A2 (A24 )/adenocarcinoma, in 2 out of 4 patients with HLA-A24/squamous cell carcinoma, and 1 of 2 patients with HLA-A2/squamous cell carcinoma. The cytotoxic activity was observed to kill PC-9 selectively, not other NSCLC lines, and the activity was substantially blocked by anti-MHC class I antibody, but not by anti-MHC class II antibody. The PC-9-specific CTL produced gamma-interferon in response to autologous tumor cells. These results indicated that the anti-PC-9 cytotoxicity was mediated by cytotoxic T lymphocytes that may recognize the T cell epitope(s) shared and presented by HLA-A2 and/or HLA-A24-positive NSCLC. PMID- 9330607 TI - Transport mechanisms of idarubicin, an anthracycline derivative, in human leukemia HL60 cells and mononuclear cells, and comparison with those of its analogs. AB - Transport mechanisms of idarubicin (IDA) in HL60 cells, as leukemia cells, and human mononuclear cells (MNCs), as normal cells, were investigated, and compared with those of its analogs. The uptake of IDA by both cell types was temperature- and concentration-dependent, was inhibited competitively by daunorubicin (DNR) and noncompetitively by adriamycin (ADR), and was stimulated by preloading of the cells with DNR and ADR, indicating the partial involvement of a carrier-mediated mechanism. On pretreatment of the cells with 2,4-dinitrophenol, IDA uptake by HL60 cells increased, but that by MNCs decreased, suggesting that IDA was partially taken up into HL60 cells via an energy-independent carrier system, and into MNCs via an energy-dependent one. We speculated that in HL60 cells the carrier concerned with IDA uptake was common to DNR and ADR, and that the binding site of IDA on the carrier was the same as that for DNR, but not that for ADR, while in MNCs the carrier system consisted of, at least in part, a carrier for DNR uptake and one for ADR uptake, and the binding site of IDA was identical to that for DNR in the former, but different from that for ADR in the latter. It appeared that the uptake of IDA was greater than those of pirarubicin, DNR and ADR in both HL60 cells and MNCs, and that IDA was incorporated into MNCs more efficiently than into HL60 cells because of the higher uptake efficacy of the carrier(s). PMID- 9330609 TI - Augmentation in chemosensitivity of intratumor quiescent cells by combined treatment with nicotinamide and mild hyperthermia. AB - C3H/He and Balb/c mice bearing SCC VII and EMT6/KU tumors, respectively, received continuous administration of 5-bromo-2'-deoxyuridine (BrdU) for 5 days using implanted mini-osmotic pumps to label all proliferating (P) cells. Nicotinamide was administered intraperitoneally before cisplatin injection and/or tumors were locally heated at 40 degrees C for 60 min immediately after cisplatin injection. The tumors were then excised, minced and trypsinized. The tumor cell suspensions were incubated with cytochalasin-B (a cytokinesis-blocker), and the micronucleus (MN) frequency in cells without BrdU labeling (quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. The MN frequency in total (P+Q) tumor cells was determined from tumors that had not been pretreated with BrdU labeling. The sensitivity to cisplatin was evaluated in terms of the frequency of induced micronuclei in binuclear tumor cells (MN frequency). In both tumor systems, the MN frequency in Q cells was lower than that in the total cell population. Nicotinamide treatment elevated the MN frequency in total SCC VII cells. Mild heating raised the MN frequency more markedly in Q cells than in total cells. The combination of nicotinamide and mild heat treatment increased the MN frequency more markedly than either treatment alone. In total SCC VII cells, nicotinamide increased 195mPt-cisplatin uptake. Mild heating elevated 195mPt-cisplatin uptake in total EMT6/KU cells. Cisplatin-sensitivity of Q cells was lower than that of total cells in both tumor systems. Nicotinamide sensitized tumor cells including a large acutely hypoxic fraction, such as those of SCC VII tumors, through inhibition of the fluctuations in tumor blood flow. Tumor cells including a large chronically hypoxic fraction such as Q cells were thought to be sensitized by mild heating through an increase in tumor blood flow. PMID- 9330608 TI - Antitumor effect of DX-8951, a novel camptothecin analog, on human pancreatic tumor cells and their CPT-11-resistant variants cultured in vitro and xenografted into nude mice. AB - DX-8951 is a novel water-soluble derivative of camptothecin. We evaluated the effects of DX-8951 on the growth of several pancreatic tumor cell lines in vitro and in vivo. In vitro cytotoxic activity of DX-8951 against SUIT-2 and KP-1N cells, as indicated by IC50 value, was several times more potent than that of SN 38, an active metabolite of CPT-11, and dozens of times more potent than that of SK&F104864 (topotecan). DX-8951 also showed the greatest cytotoxicity against CPT 11-resistant variants, SUIT-2/CPT-11 and KP-1N/CPT-11 cells, and the cross resistance of these cells to DX-8951 was lower than that to SN-38 and SK&F104864. Topoisomerase I inhibitory activity of DX-8951 was about three-fold stronger than that of SN-38, as measured in crude nuclear extract obtained from SUIT-2 cells. DX-8951 induced DNA fragmentation, a specific feature of apoptosis, in SUIT-2 cells more effectively than SN-38. DX-8951 exhibited potent antitumor effects against SUIT-2 in a solid tumor model and in a liver metastasis model, in which tumor cells were xenografted subcutaneously and intrasplenically, respectively, into nude mice. The in vivo effects were closely similar to or somewhat superior to those of CPT-11. DX-8951 also showed significant antitumor effects against SUIT-2/CPT-11 solid tumors, against which CPT-11 had no effect. These results suggest that, on the basis of its strong antitumor activity and effectiveness against CPT-11-resistant tumors, DX-8951 may be a useful therapeutic agent in the treatment of human cancer. The potent cytotoxicity of DX-8951 may result from strong inhibition of topoisomerase I, which may then trigger apoptotic cell death. PMID- 9330610 TI - Effectiveness of mammographic screening for breast cancer in women aged over 50 years in Japan. AB - The optimal age for effective screening of subjects for breast cancer by mammography in Japan was studied based on the results of two mammographic screening systems (systems I and II) in Tokushima Prefecture. System I consisted of visit screening using a bus equipped with a mammographic apparatus. System II consisted of central screening performed at Tokushima Health Screening Center. The examinees numbered 2,500 and 3,707 in systems I and II, respectively. There was a significant difference between the two screening systems in the age distribution of the examinees. The detection rates of breast cancer were 0.6% and 0.24% in systems I and II, respectively, which are 2-5 times higher than that (0.12%) obtained by conventional screening using physical examination alone. The detection rate increased especially in the sixth and seventh decades of life. The sensitivity of mammography screening was 93.3% in system I and 81.1% in system II. Higher sensitivity (100%) than that (73%) of screening by physical examination was obtained in women aged over 50. The proportion of stage I was 60% in system I and 66.7% in system II, compared with 32-65% in the United States and Europe. The rates of no nodal involvement were high, being 77.8% and 83.3% in systems I and II, respectively, compared with 57-71% in other countries. Breast conserving therapy was applied to 18 of the 24 patients with breast cancer detected by the two screening systems. In addition, in Wolfe's classification of mammograms, the proportion of DY (mammary dysplasia) pattern was remarkably low, being 3.2% in the sixth decade and 0.8% in the seventh decade, compared with 16.6% in women aged 49 years. These results indicate that mammographic screening is effective in women aged over 50 years in Japan, as has been found in other countries. PMID- 9330611 TI - Developmental expression of a DNA repair gene in Arabidopsis. AB - Exposure to alkylating agents results in the formation of a wide variety of DNA damage products. One of these, 3-methyladenine (3-mAde), is lethal if left unrepaired. The 3-methyladenine glycosylase (aMAG) gene of Arabidopsis thaliana is required for base excision repair of this lesion, and probably shares the ability of other 3-mAde glycosylases to recognize and excise a broad spectrum of damaged bases. Given the fact that DNA damage products can act as blocks to both DNA and RNA synthesis, one would expect that this protein should be expressed to some degree in all living cells. Using a DIG-labeled aMAG antisense RNA as a probe, we have investigated the developmental and tissue-specific expression of this repair gene. We found that the gene is preferentially expressed in meristematic tissue, the developing embryo and endosperm, and organ primordia. This pattern of expression is consistent with a requirement for expression in rapidly dividing tissues. However, high levels of expression were also observed in growing leaves, a tissue that is undergoing a relatively low rate of cell division. This result suggests that 3-mAde glycosylase is required not only for DNA replication, but also for cell growth. PMID- 9330612 TI - The roles of the eukaryotic DNA polymerases in DNA repair synthesis. PMID- 9330613 TI - Identification of defective illegitimate recombinational repair of oxidatively induced DNA double-strand breaks in ataxia-telangiectasia cells. AB - Ataxia-telangiectasia (A-T) is an autosomal-recessive lethal human disease. Homozygotes suffer from a number of neurological disorders, as well as very high cancer incidence. Heterozygotes may also have a higher than normal risk of cancer, particularly for the breast. The gene responsible for the disease (ATM) has been cloned, but its role in mechanisms of the disease remain unknown. Cellular A-T phenotypes, such as radiosensitivity and genomic instability, suggest that a deficiency in the repair of DNA double-strand breaks (DSBs) may be the primary defect; however, overall levels of DSB rejoining appear normal. We used the shuttle vector, pZ189, containing an oxidatively-induced DSB, to compare the integrity of DSB rejoining in one normal and two A-T fibroblast cells lines. Mutation frequencies were two-fold higher in A-T cells, and the mutational spectrum was different. The majority of the mutations found in all three cell lines were deletions (44-63%). The DNA sequence analysis indicated that 17 of the 17 plasmids with deletion mutations in normal cells occurred between short direct repeat sequences (removing one of the repeats plus the intervening sequences), implicating illegitimate recombination in DSB rejoining. The combined data from both A-T cell lines showed that 21 of 24 deletions did not involve direct-repeats sequences, implicating a defect in the illegitimate recombination pathway. These findings suggest that the A-T gene product may either directly participate in illegitimate recombination or modulate the pathway. Regardless, this defect is likely to be important to a mechanistic understanding of this lethal disease. PMID- 9330614 TI - Biochemical and physicochemical characterization of normal and variant forms of human MTH1 protein with antimutagenic activity. AB - 8-Oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP) is produced during cellular metabolism, and its misincorporation into DNA causes mutation. Human cells possess an enzyme that hydrolyzes 8-oxo-dGTP to the corresponding nucleoside monophosphate, thereby preventing misincorporation of 8-oxo-7,8 dihydroguanine into DNA. Sequence analyses of the MTH1 gene, encoding the 8-oxo 7,8-dihydro-2'-deoxyguanosine 5'-triphosphatase (8-oxo-dGTPase) protein in human cell lines revealed that a G to A base substitution frequently occurs at codon 83, which causes a change of valine to methionine in the MTH1 protein [Wu, C. et al., Biochem. Biophys. Res. Commun. 214 (1995) 1239-1245]. Here we isolated cDNAs for the two types of MTH1 protein and expressed them in Escherichia coli mutT-. cells, devoid of their own 8-oxo-dGTPase activity. The two forms of proteins were purified to physical homogeneity, and amino acid analyses confirmed that the variant protein, Met83-MTH1, indeed carries the corresponding amino acid substitution. Met83-MTH1, but not normal type Val83-MTH1, was separated into two peaks in hydrophobic interacting chromatography. 8-Oxo-dGTPase activity of Met83 MTH1 is more thermolabile than that of Val83-MTH1. Circular dichroism (CD) and fluorescence spectroscopic analyses confirmed this conclusion. CD further indicated that Met83-MTH1 has a higher alpha-helix content. PMID- 9330615 TI - Characterization of a human homolog of (6-4) photolyase. AB - (6-4)Photolyase catalyzes light-dependent repair of UV-induced pyrimidine (6-4) pyrimidone photoproducts. A human cDNA clone which has high sequence homology to the (6-4)photolyase gene (H64PRH gene) was identified. In this paper we also isolated a genomic clone corresponding to the H64PRH cDNA and mapped it to chromosome 12q24.1 by fluorescence in situ hybridization (FISH). Northern-blot analysis revealed transcription of this gene in all human tissues examined. The H64PRH protein was overproduced in E. coli, partially purified and characterized. Like (6-4)photolyase, the enzyme contains two chromophores, one of which is FAD. However, the enzyme does not show any detectable photolyase activity. PMID- 9330616 TI - Cell cycle-dependent protein expression of mammalian homologs of yeast DNA double strand break repair genes Rad51 and Rad52. AB - Recently, human and rodent homologs of yeast repair genes Rad51 and Rad52 have been identified and proposed to play roles in DNA double-strand break (DSB) repair. In this study, cell cycle-dependent expression of human and rodent RAD51 and RAD52 proteins was monitored using two approaches. First, flow cytometric measurements of DNA content and immunofluorescence were used to determine the phase-specific levels of RAD51 and RAD52 protein expression in irradiated and control populations. The expression of both proteins was lowest in G0/G1, increased in S and reached a maximum in G2/M. No difference was found in the whole-cell level of RAD51 or RAD52 protein expression between gamma-irradiated and control cell populations. Second, cell cycle-dependent protein expression was confirmed by Western analysis of populations synchronized in G0, G1 and G2 phases. Analysis of V3, a hamster equivalent of SCID, indicates that the protein level increases of RAD51 and RAD52 from G0 to G1/S/G2 do not require DNA-PK. PMID- 9330617 TI - Oxidative phosphorylation dysfunction does not increase the rate of accumulation of age-related mtDNA deletions in skeletal muscle. AB - Several reports described an age-related accumulation of a particular mitochondrial DNA (mtDNA) deletion ('common deletion') in post-mitotic tissues. These findings led to the hypothesis that free radicals generated inside the mitochondria could damage mtDNA during a normal life span. The impaired electron transfer function resulting from mtDNA damage would increase the production of free radicals creating a vicious cycle. If this vicious cycle is an important player in the somatic accumulation of mtDNA deletions, patients with impaired oxidative phosphorylation (regardless of the primary defect) should have an accelerated accumulation of mtDNA deletions. We tested this hypothesis by performing three analyses: (a) comparing the amounts of the mtDNA 'common deletion' in normal controls and patients with genetically characterized mitochondrial disorders associated with pathogenic mtDNA point mutations or deletions other than the common deletion; (b) analyzing the co-segregation of the age-related mtDNA common deletion with a pathogenic mtDNA point mutation; and (c) by the detection of multiple mtDNA deletions by long PCR in controls and patients with mitochondrial disorders. We observed a positive correlation between age and common deletion levels in controls (r = 0.80) and patients (r = 0.69). The slopes of the curves were similar, suggesting that the rate of accumulation of the age related common deletion was the same in both groups. We could not find a co segregation of the pathogenic point mutated mtDNA molecules with the common deletion nor increased number of age-related deletions in patients. Our data do not support the hypothesis that a vicious cycle (damage to mtDNA would affect the respiratory function, leading to the generation of more free radicals, which in turn would provoke additional mtDNA damage) is an important factor in the accumulation of age-related mtDNA deletions. PMID- 9330618 TI - Spontaneous mutation frequencies and spectra in p53 (+/+) and p53 (-/-) mice: a test of the 'guardian of the genome' hypothesis in the Big Blue transgenic mouse mutation detection system. AB - TSG-p53/Big Blue double transgenic mice offer a powerful tool for examining the effect of a p53 germline mutation on spontaneous somatic mutation in vivo. After sequencing the DNA-binding domain of the lacI gene, we previously reported no differences in mutant frequency between p53 nullizygous (-/-) and p53 wild-type (+/+) mice in liver, spleen and brain. However, jackpot mutations elsewhere in the gene may have obscured a real difference in mutation frequency and the small sample size of mutations not at CpG dinucleotides (n = 23) may have been insufficient to reveal differences in mutation spectra. Herein we have sequenced the entire lacI gene, including the promoter and lacZ operator regions. 123 additional independent mutations have been found including 70 mutations not at CpG sites. The mutation frequency was determined by correcting for jackpot mutations. There were no statistically significant differences in mutation frequency or spectrum between the p53 (+/+) and p53 (-/-) genotypes in any of the three tissues. PMID- 9330619 TI - The inhibition by flavonoids of 2-amino-3-methylimidazo[4,5-f]quinoline metabolic activation to a mutagen: a structure-activity relationship study. AB - The mutagenicity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in Salmonella typhimurium TA98 is inhibited by flavonoids with distinct structure antimutagenicity relationships (Edenharder, R., I. von Petersdorff I. and R. Rauscher (1993). Antimutagenic effects of flavonoids, chalcones and structurally related compounds on the activity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and other heterocyclic amine mutagens from cooked food, Mutation Res., 287, 261 274). With respect to the mechanism(s) of antimutagenicity, the following results were obtained here. (1) 7-Methoxy- and 7-ethoxyresorufin-O-dealkylase activities in rat liver microsomes, linked to cytochrome P-450-dependent 1A1 and 1A2 monooxygenases catalyzing oxidation of IQ to N-hydroxy-IQ (N-OH-IQ), were effectively inhibited by 16 flavonoids (IC50: 0.4-9.8 microM). Flavones and flavonols are in general more potent enzyme inhibitors than flavanones, isoflavones, and chalcones. Among flavones the presence of hydroxyl or methoxyl groups resulted in minor changes only. However, among flavonols and flavanones the parent compounds exerted the strongest inhibitory effects, which decreased in dependence on number and position of hydroxyl functions. Contrary to the results obtained in the Salmonella assay in the tests with alkoxyresorufins no extraordinary counteracting effects of isoflavones, of hydroxyl groups at carbons 6 or 2' or of the elimination of ring B (benzylideneacetone) were detected. (2) No effects of flavonoids on NADPH-dependent cytochrome P-450 reductase activity could be detected. (3) The effects of 30 flavonoids on mutagenicity induced by N OH-IQ in S. typhimurium TA98NR were again structure dependent. The most striking feature was the, in principle, reverse structure-antimutagenicity pattern as compared to IQ: non-polar compounds were inactive and a 50% inhibition was achieved only by some flavones and flavonols (IC50: 15.0-148 nmol/ml top agar). Within the flavone and flavonol subgroups inhibitory effects increased in dependence on number and position of hydroxyl functions. Isoflavones and flavanones, however, as well as glycosides, were inactive. Hydroxyl groups at carbons 7, 3', 4', and 5' generated antimutagenic compounds, a hydroxyl function at C5 was ineffective, but hydroxyls at C3 and 6 as well as methoxyl groups at C3' (isorhamnetin) or 4' (diosmetin) generated comutagenic compounds. 4. Cytosolic activation of IQ to mutagenic metabolites as determined by experiments with the hepatic S105 fraction comprises about 10% of the mutagenicity after activation by the combined microsomal and cytosolic fractions (S9). The pattern of inhibition as produced by 20 flavonoids was closely similar to that observed with the S9 fraction. 5. In various experiments designed for modulation of the mutagenic response, it could be shown that further mechanisms of flavonoid interaction with the overall mutagenic process may exist, such as interactions with biological membranes (luteolin, fisetin) and effects on fixation and expression of.DNA damage (flavone, fisetin). PMID- 9330621 TI - Characterization of DNA adducts in Chinese hamster ovary cells treated with mutagenic doses of 1- and 3-nitrosobenzo[a]pyrene and the trans-7,8-diol-anti 9,10-epoxides of 1- and 3-nitrobenzo[a]pyrene. AB - The environmental contaminants 1- and 3-nitrobenzo[a]pyrene (1- and 3-nitro-BaP) are mutagens in Chinese hamster ovary (CHO) cells with exogenous metabolic activation. Previous studies demonstrated the potent direct-acting mutagenicity of the oxidized metabolites, trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10 tetrahydro-1-nitrobenzo[a] pyrene (1-NBaPDE) and trans-7,8-dihydroxy-anti-9,10 epoxy-7,8,9, 10-tetrahydro-3-nitrobenzo[a]pyrene (3-NBaPDE), and the partially nitroreduced metabolites, 1- and 3-nitrosobenzo[a]pyrene (1- and 3-NO-BaP). In this study, we have identified the major adduct formed by incubation of calf thymus DNA with 1-NBaPDE and used this standard in conjunction with other adduct standards to characterize the 32P-postlabeled DNA adducts produced by 1- and 3 nitro-BaP metabolites in CHO cultures. The major adduct from 1-NBaPDE exposure was 10-(deoxyguanosin-N2-yl)-7,8,9-trihydroxy-7,8,9,10-tetrahydro-1- nitrobenzo[a]pyrene; from 3-NBaPDE, 10-(deoxyguanosin-N2-yl)-7,8,9-trihydroxy 7,8,9,10-tetrahydro-3- nitrobenzo[a]pyrene; from 1-NO-BaP, 6-(deoxyguanosin-N2 yl)-1-aminobenzo[a]pyrene; and from 3-NO-BaP, 6-(deoxyguanosin-N2-yl)-3 aminobenzo[a]pyrene. For comparison, the adducts formed by trans-7,8-dihydroxy anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and the related nitroreduced derivative 6-nitrosobenzo[a]pyrene were also examined. The nitrobenzo[a]pyrene DNA adducts described in this study are proposed to be involved in the mutagenicity of 1- and 3-nitro-BaP upon either oxidative or reductive metabolism. PMID- 9330620 TI - DNA demethylation and pericentromeric rearrangements of chromosome 1. AB - Rearrangements in the vicinity of the centromere of chromosome 1 are over represented in many types of human cancer and are a characteristic feature of a rare genetic disease called ICF (immunodeficiency, centromeric heterochromatin instability, and facial anomalies). Evidence is presented that implicates DNA hypomethylation in the formation of these pericentromeric chromosomal anomalies. The DNA methylation inhibitors 5-azadeoxycytidine and 5-azacytidine, but not other tested genotoxins, induced the preferential formation of pericentromeric rearrangements of chromosome 1 at a very high frequency in a pro-B-cell line (FLEB14) and at a lower frequency in a mature B-cell line (AHH-1). These abnormal chromosomes appear identical to the diagnostic chromosomal aberrations in the ICF syndrome. A major component of the pericentromeric DNA in chromosome 1, satellite 2, was shown to be hypomethylated in an ICF B-cell line, although DNA from this cell line did not display detectable overall hypomethylation. It is hypothesized that demethylation in certain DNA regions, including in pericentromeric satellite DNA, helps lead to pericentromeric chromosomal rearrangements in lymphocytes from ICF patients and in normal lymphoblastoid cells incubated in vitro with DNA demethylating agents. PMID- 9330622 TI - Predisposing genes and increased chromosome aberrations in lung cancer cigarette smokers. AB - Genotoxic effects linking cigarette smoking with lung cancer have not been consistently demonstrated, therefore claims for the cause-effect relationships are vigorously contested. Using matched populations of 22 lung cancer patients who have been cigarette smokers (LCP), 22 non-cancerous cigarette smokers (SC) and 13 non-smokers (NSC), we have applied the fluorescence in situ hybridization (FISH) tanden probe assay to elucidate the frequency of chromosome breakage among the participants. Two probes were used, a classical satellite probe which hybridizes to the large heterochromatin region of chromosome 1, and an alpha satellite probe which targets a small region adjacent to the heterochromatin probe. The highest frequency of structural aberrations was observed in LCP (1.4 +/- 0.1) followed by SC (1.25 +/- 0.1) and NSC (0.4 +/- 0.1). Aberration frequencies were not significantly different between LCP and SC (p > 0.05), however, a statistically significant difference was detected between the smoker populations combined (LCP and SC) and the NSC (p < 0.001). The breakage frequencies showed a positive correlation with duration of smoking for LCP (r = 0.5; p < 0.01), but not for SC (P > 0.05). In addition, the aberration frequencies were influences by the inheritance of polymorphic glutathione S transferase (GST) genes. LCPs missing one or the other GST (GSTM1 or GSTT1) genes were found to have significantly higher chromosome breaks compared to LCPs with both genes present (p < 0.05). Our data indicate that genetic predisposition and chromosome aberrations may be mechanistically related to the initiation of lung carcinogenesis; therefore, they may be useful biomarkers for lung cancer among cigarette smokers. PMID- 9330624 TI - Somatic mutation of the glucose-6-phosphate dehydrogenase (g6pd) gene in colonic stem cells and crypt restricted loss of G6PD activity. AB - The study of somatic mutation frequency, particularly stem cell somatic mutation, is important to the understanding of mechanisms of carcinogenesis. The models currently in use for studies in stem cell tissues such as the colon infer the presence of stem cell somatic mutation from alteration in enzyme function, when this has shown to be mutagen dose dependent, restricted to the unit of clonal architecture, and persistent. The present study identifies and characterises somatic mutations in the g6pd gene in individual mouse colonic crypts showing histochemically demonstrable loss of G6PD activity. Microdissection of single crypts, showing either normal or low G6PD activity by histochemistry was performed in mice treated with ethylnitrosourea (ENU), and the presence of point mutations sought by PCR and direct sequencing. Because of the limitation of the small amount of partially degraded (due to fixation) DNA available from each crypt, only about 20% of the coding region of the g6pd gene could be sequenced. Despite this, somatic mutations were identified in 3 of the 9 crypts analysed which showed loss of G6PD activity, but in none of the crypts with normal activity. Each of the mutations identified would be predicted to lead to a decrease in enzyme activity. We conclude that we have confirmed that the crypt restricted loss of G6PD activity is indeed due to stem cell somatic mutation in the g6pd gene, and suggest that the G6PD model can be used as a paradigm for other models where somatic mutation is inferred from a change in histochemically identifiable gene expression. PMID- 9330623 TI - Analysis of mutations in the K-ras and p53 genes of lung tumors and in the hprt gene of 6-thioguanine-resistant T-lymphocytes from rats treated with 1,6 dinitropyrene. AB - Direct pulmonary instillation of 1,6-dinitropyrene (DNP) into male Fischer 344 rats results in a dose-dependent induction of lung tumors and 6-thioguanine resistant (TGr) T-lymphocytes. The treatment also results in DNP binding to dG in the lung and in T-lymphocytes. In the present study, we have examined the types of mutations associated with these responses to DNP. Sequencing of DNA amplification products from 20 DNP-induced lung tumors identified 5 mutations in K-ras codon 12, 4 GGT-->TGT transversions and one GGT-->GAT transition. No mutations were found in K-ras codons 13 or 61. Single-strand conformation polymorphism analysis of p53 exons 5-8 revealed mobility shifts indicative of mutation in 9 of the 20 tumor samples. Eight of the mutations were substitutions at G:C base pairs, and one was a deletion of a single G:C base pair. DNA from 161 TGr lymphocyte colonies cultured from DNP-treated rats was examined for point mutations by amplification of hprt exons 2, 3, and 8, and screening the products for mutant: wild-type heteroduplex formation by denaturing gradient-gel electrophoresis. Only three mutations were found, a G-->T transversion in exon 3, a G-->A transition in exon 8, and a complex mutation consisting of a tandem G-->T transversion and a one base deletion in exon 3. The mutations identified in the DNP-induced lung tumors and TGr T-lymphocytes are consistent with the formation of dG-DNA adducts by DNP. The extremely low recovery of point mutations from TGr lymphocytes suggests that DNP induces a substantial number of mutations by other mechanisms. PMID- 9330626 TI - Chromosomal aberrations in vitro induced by aneugens. AB - Various aneugens were reported to induce structural chromosomal aberrations beside their influence on cell division and their aneugenic potential To asses, whether a relationship between disturbance of cell division and clastogenic potential exists, CHO cells were treated with the well-known aneugens colcemid, colchicine and vincristine and investigated for the induction of structural chromosomal aberrations, polyploid cells and alterations in mitotic index. At low and intermediate concentration, all compounds induced polyploidy and an increase in mitotic index, but no structural aberrations at all. However, at high concentrations, colcemid and colchicine both induced numerous structural chromosomal aberrations in diploid cells. Colchicine was also clastogenic in tetraploid cells. Vincristine did not induce structural chromosomal aberrations in diploid cells, but in tetraploid cells. The clastogenic effects showed a clear cut threshold with all three compounds. Furthermore, it was found that the tetraploid condition in CHO cells is generally accompanied by an increase in structural chromosomal aberrations, in vehicle controls as well as in cultures treated with the aneugens. Nevertheless, this study demonstrates that for the three aneugenic compounds tested, no direct relationship between compound induced disturbance of cell cycle and compound induced structural chromosomal aberration incidence exists. PMID- 9330625 TI - On the recovery of single spots with the flr phenotype in the wing spot test in Drosophila. AB - Graf et al. (U. Graf, F.E. Wurgler, A.J. Katz, H. Frei, H. Juon, C.B. Hall, P.G. Kale, Somatic mutation and recombination test in Drosophila, Environment Mutagen. 6 (1984) 153-188.) described the overall results of assays of a series of compounds in the Drosophila wing spot test as indicating that single mwh spots appeared most frequently, followed by less frequent twin spots with both mwh and flr cells and lastly the 'quite rare' single flr spots. Data are presented below demonstrating that some compounds behave in a manner consistent with the above description, whereas others do not in that the frequency of single flr spots is equal to or exceeds that of twin spots and cannot be described as occurring 'rarely'. It is suggested that (large) flr singles be used as a measure of mutations/deletions directly from treated transheterozygotes. An argument is presented questioning the use of treated mwh +/+ TM3 individuals as an assay of mutations/deletions at the mwh+ locus. PMID- 9330627 TI - Naming the mutagenic nucleic acid base analogs: the Galatea syndrome. PMID- 9330628 TI - Response to "Selecting chemicals and assays for assessing mammalian germ cell mutagenicity" [M.D. Shelby, Mutation Research, 352 (1996) 159-167]. PMID- 9330629 TI - Structural and developmental analysis of the mouse peripherin/rds gene. AB - Mutations in the peripherin/rds gene have been reported to be associated with different forms of human autosomal dominant retinitis pigmentosa (ADRP) and macular degeneration (MD). To better understand the disruptive role of these mutations, knowledge of the structure-function relationship of the peripherin/rds gene is needed. To facilitate that, genomic clones encoding the mouse gene were isolated using bovine cDNA sequences as probes. Sequence analysis of clone lambda 6-1-1, that contained the entire coding sequence for the mouse peripherin/rds, yielded the exon-intron organization of the gene. The gene is composed of three exons (581, 247, and 213 bp) and two introns with the first and second introns 8.6 kb and 3.7 kb in size, respectively. Two major (1.6 and 2.7 kb) and three minor (4.0, 5.5, 6.5 kb) transcripts were detected on RNA blots. The major transcripts first appeared in the brain at embryonic day 13 and in the retina at postnatal day 1. Transcripts were missing in brain and eye of mice at embryonic day 15. Several transcription start sites were mapped within 26 nucleotides approximately 200 bp upstream from the translation initiation site. However, transcripts varied in the lengths of their 3' untranslated portion as a result of the utilization of different polyadenylation signals. PMID- 9330630 TI - Selective loss of the hepatic phenotype due to the absence of a transcriptional activation pathway. AB - Liver-enriched trans-acting factors hepatocyte nuclear factor-1 alpha (HNF1 alpha) and -4 (HNF4) are components of a transcriptional activation pathway that is thought to play a major role in hepatic gene activation. We previously described the isolation and characterization of distinct classes of hepatoma variants which lack the HNF4-->HNF1 alpha pathway (1). In order to determine the influence of the HNF4-->HNF1 alpha pathway on hepatic gene expression, genetic rescue experiments were done using hepatoma variant line H11 as a model system. Results suggest that this pathway is required for basal expression of a number of endogenous hepatocyte-specific genes. Complementation groups were established by fusion of H11 cells with other variant lines. Lastly, introduction of human chromosome 20 (containing the HNF4 locus) or randomly-marked human chromosomes into H11 cells failed to rescue the hepatic phenotype, suggesting that what appears to be a 'simple' defect may involve multiple genetic loci. PMID- 9330631 TI - High-efficiency retroviral infection of primary myoblasts. AB - In the past, it has been hard to introduce genes into primary myoblasts without selection, as they have been very difficult to transfect or infect. We describe conditions under which mouse primary skeletal muscle myoblasts can be infected with retroviral vectors at high efficiency. Infection can be greatly increased by minimizing the time during which cells are exposed to virus, adding a minimal centrifugation step, and supplementing the infection cocktail to mimic more closely primary myoblast growth medium. Under these conditions, one round of exposure to virus results in an infection efficiency of up to 80%, whereas 4-5 rounds of infection over a two day period reproducibly yield an infection efficiency of > 99%. These methods greatly enhance the potential for studying genetically engineered primary myoblasts from any mouse strain, transgenic or knockout, and may have useful application to other primary cell types that are refractory to transfection or infection. PMID- 9330632 TI - Chromosomal aberrations induced by defined DNA double-strand breaks: the origin of achromatic lesions. AB - The mechanisms of formation of chromosomal aberrations are poorly understood, despite the common use of aberrations as a measure of the genetic effects of physical and chemical agents. We have used restriction endonucleases to introduce defined DNA double-strand breaks into mammalian cells, and measured chromosomal aberration formation relative to the activity of the endonuclease. The endonucleases AluI and Sau3AI remain active for a relatively short time under simulated cellular conditions and induce achromatic lesions ('gaps') in chromatids only within the first hour or two following treatment. In contrast, the endonuclease MboI (an isoschizomer of Sau3AI) is active for an extremely long time and continues to produce chromatid gaps during the whole 12 hr sampling period. This observation strongly suggests that the aberrations classified as gaps are a manifestation of unrejoined DNA double-strand breaks. The formation of gaps may relate to the opportunities for repair of DNA breaks in relation to cell cycle position. It is more difficult to relate the formation of structural chromatid aberrations to the endonuclease activity, although at relatively low concentrations all 3 endonucleases gave similar levels of structural aberrations. PMID- 9330633 TI - Assignment of the human gene encoding eukaryotic initiation factor 4E (EIF4E) to the region q21-25 on chromosome 4. AB - We recently cloned genomic sequences containing the promoter region for the messenger RNA cap binding protein (eIF4E). As the rate-limiting step in translation, eukaryotic initiation factor 4E is important in cellular growth control. Using oligonucleotide primers specific for the promoter region in polymerase chain reactions (PCR), we amplified the human gene in a chromosome 4 specific human/rodent somatic cell panel. This panel mapped single copy genomic sequences for eIF4E in the region 4q21 to 4q25. PMID- 9330634 TI - Assignment of the possible HTLV receptor gene to chromosome 17q21-q23. AB - We have determined the HTLV (Human T-cell leukemia virus) receptor localization more precisely on the human chromosome 17. Based on the fact that HTLV infection induces syncytium formation of infected cells as a result of interaction between the viral envelope and viral receptor, we performed the sensitive biological assay using recombinant vaccinia expression system. Our results from the induced syncytium pattern of the somatic hybrid cell lines with different deletions indicated that the HTLV receptor gene may reside from q21 to q23 on the long arm of the human chromosome 17. PMID- 9330635 TI - Chromosomal localization to 19q13.3, partial genomic structure and 5' cDNA sequence of the human symplekin gene. AB - Exon trapping from cosmids mapping to chromosome 19q13.3 yielded 6 exonic sequences that matched the human symplekin gene, which encodes a tight junction related protein. One exonic sequence identified a 4.0 kb brain cDNA clone, R6E1, which contained 302 bp 5' to the originally reported 3.7 kb symplekin cDNA. A portion of this novel 5' sequence matched an additional trapped exonic sequence which was obtained from the most telomeric cosmid analyzed. The symplekin gene thus lies in a telomeric-to-centromeric direction on 19q13.3. Only three cosmids from a large 19q13.3 contig hybridized with R6E1, thereby assigning the symplekin gene to a 40 kb region immediately telomeric to gene 59 and the DM protein kinase gene. The 5' end of the R6E1 clone has a potential initiation codon with a strong Kozak sequence and Northern blot analysis detected a 4.2 kb signal in most human tissues, indicating that R6E1 may be a complete cDNA sequence. Based on the trapped exonic sequences, twelve exon-intron boundaries were predicted. PMID- 9330636 TI - Localization of the candidate tumor suppressor gene ING1 to human chromosome 13q34. AB - A novel gene ING1 was recently cloned and defined as a candidate tumor suppressor gene. Reduced expression and rearrangements of ING1 are found in several tumor cell lines, ING1 overexpression is associated with cell growth arrest and ING1 suppression promotes neoplastic transformation (1). Using radiation hybrid mapping technique ING1 was assigned to subtelomeric region of the long arm of human chromosome 13 (13q34) which is known to be frequently rearranged in squamous carcinomas of head and neck. PMID- 9330637 TI - Isolation of human ear specific cDNAs and construction of cDNA libraries from surgically removed small amounts of inner ear tissues. AB - We have used representational difference analysis (RDA) for subtractive hybridization of oligo dT primed directionally cloned cDNA libraries from human inner ear tissue and a B-lymphoblast cell line. Two rounds of subtraction amplification, followed by differential hybridization of selected clones led to the isolation of genes which were specific to the ear. Sequence analysis of randomly chosen clones revealed the presence of a histidine rich Ca2+ binding protein, human dynamin, collagen type 1A1, collagen type 2A1, SPARC, human growth hormone, and several specific genes which had no sequence homology in the data base. Furthermore, to apply these techniques for isolating genes specific to distinct inner ear structures and/or cell types of inner ear for which the starting tissue material is limiting, we have used a modified PCR based protocol to construct representative cDNA libraries. We have characterized a cDNA library constructed from small amounts of inner ear tissues recovered by ablative surgical procedure involving labyrinthectomy. The potential application of these protocols for isolating genes involved in hearing and deafness is discussed. PMID- 9330638 TI - Replication timing properties of the human HPRT locus on active, inactive and reactivated X chromosomes. AB - X chromosome inactivation is associated with a highly asynchronous pattern of DNA replication at most X-linked loci in females. We studied the human HPRT locus, which is subject to X inactivation and expressed from only the active homolog, with the goal of comparing replication properties between the active and inactive homologs in this region using a fluorescence in situ hybridization approach. We found that in normal female lymphoblasts this locus is replicated in a highly asynchronous manner across a broad, discrete 500-600 kb zone with earliest replication appearing at the gene coding sequence. This general timing profile is maintained in normal male lymphoblasts, as well as in hamster x human hybrid cells containing the active human X chromosome. However, the inactive human X chromosome in the hamster cell background does not appear to function in a fully equivalent manner to the normal inactive X chromosome in female cells. Furthermore, reactivation of the inactive human X chromosome in a hamster x human hybrid system by 5-azacytidine treatment and HAT selection restores early replication at the HPRT gene itself, but does not change the overall domain behavior. PMID- 9330639 TI - A cell line selected for resistance to ionizing radiation exhibits cross resistance to other genotoxic agents and a mutator phenotype for loss of heterozygosity events. AB - An ionizing radiation resistant derivative was obtained from the mouse P19H22 (aprt hemizygote) embryonal carcinoma cell line by repeated exposure to 137Cs gamma radiation. Ionizing radiation resistance in the 6Gy-R cell line was not correlated with a failure to undergo cell cycle arrest or a loss of the p53 response after exposure to 137Cs gamma radiation. Moreover, the cells did not display increased resistance to bleomycin, a double strand break inducing agent. However, the cells did display increased resistance to ultraviolet radiation, ethyl methanesulfonate, and 95% oxygen. A mutational analysis demonstrated a > 700 fold-fold increase in the frequency of aprt mutants for the 6Gy-R cells, but no change in the frequency of hprt or dhfr mutants. A molecular analysis suggested that the aprt mutations in the 6Gy-R cells arose by recombinational events. A possible association between radiation resistance, DNA repair, and a mutator phenotype for large-scale mutational events is discussed. PMID- 9330640 TI - Expression of Rap 1 suppresses genomic instability of H-ras transformed mouse fibroblasts. AB - Among the multiple genetic changes that occur during cancer progression are the activation of proto-oncogenes and the inactivation or loss of genes encoding tumor suppressors. The potential roles for these genes in the perturbation of genome stability continues to be of major interest. We have previously shown that conditional expression of H-ras in NIH3T3 cells increases genetic instability in these cells, rendering them more permissive to gene amplification and to the generation of chromosome aberrations which can be induced within a single cell cycle. In the present study we show that genetic instability induced by H-ras expression can be suppressed by co-expression of Rap 1, a Ras-related tumor suppressor gene. An NIH3T3 cell line transformed with activated human H-ras was transfected with Rap 1. Expression of the Rap 1 gene reverted the transformed cells to a flat morphology. The reverted cells reestablished contact inhibition of growth and lost the capacity to form colonies in soft agar. These cells were subsequently studied for the role of Rap 1 on the suppression of genomic instability induced by oncogenic H-ras. Cells transformed with H-ras manifest an increase in methotrexate resistance as measured by an increase in Dhfr gene amplification. Cells which concommitantly express Rap 1 showed reduced levels of methotrexate resistance as well as reduction of gene amplification capacity. Furthermore fluorescent-in-situ hybridization (FISH) with a pancentromeric mouse probe showed that elevated levels of chromosome aberrations in cells expressing H ras were also suppressed after co-expression of Rap 1. PMID- 9330642 TI - Isolation of a hamster cDNA homologous to the mouse and human cyclin kinase inhibitory protein p27Kip1. AB - We report here the isolation, cloning and sequencing of a hamster cDNA homologous to the mouse cyclin kinase inhibitory protein p27Kip1. The full length hamster cDNA sequence (p30Kip1)5 revealed 91% similarity with the previously reported mouse and human cDNAs and coded for a protein of 198 amino acids, results which were very similar to that observed for the mouse and human protein. Western blotting analysis performed using a polyclonal antibody against a mouse cyclin kinase inhibitory protein, p27Kip1, revealed the presence of a strongly reactive protein band at 30 kDa (as opposed to 27 kDa in mouse and human cells) in cell lysate prepared from rat and hamster cells. Although the size of the cyclin kinase inhibitory protein cDNA transcript is similar in mouse and hamster, it is likely that the differential mobility of the hamster p30Kip1 protein compared to the mouse and human p27Kip1 protein could be due to post-translational modifications. PMID- 9330641 TI - Map location, genomic organization and expression patterns of the human RED1 RNA editase. AB - A cDNA fragment containing sequences homologous to the rat RED1 RNA editase gene was recently identified on human chromosome 21. Here we report the location of this cDNA in distal 21q22.3 near the CD18 gene. We also report isolation of cDNA clones containing the complete coding region of the human RED1 gene, and use of this sequence to determine the genomic structure from overlapping cosmids. Human RED1 spans approximately 25 kb and is composed of 10 exons containing coding sequences. The two RNA binding domains are located within a single large, 935 nucleotide, exon 2. An alternatively processed exon 6 potentially interrupts the catalytic domain. Exon 10 is largely composed of the 3' untranslated region, which is unusually high in GC content and contains a segment that is > 90% identical with the 3' UT of the homologous rat gene. A survey of expression patterns reveals differential processing of the 5 and 8.5 kb transcripts in all sources examined. The difference in transcript size likely results from alternative processing in the 3' UT. Potential relevance of overexpression of RED1 to the development of the Down Syndrome phenotype is discussed. PMID- 9330644 TI - Rapid characterization of human chromosomes in hybrid cell lines by primed in situ (PRINS) labeling. AB - The primed in situ (PRINS) labeling technique allows a rapid and specific labeling of human chromosomes in situ. This method is based on annealing of specific oligonucleotide primers and subsequent primer extension by a Taq DNA polymerase. We have developed a PRINS protocole for the cytogenetic analysis of somatic hybrid cell lines. Painting of human chromosomes is performed using Alu specific primers. Individual human chromosomes are identified using chromosome specific alpha-satellite primers. The method was successfully tested to 3 different human-hamster hybrid cell lines. This approach provides an interesting alternative to classical cytogenetic and in situ hybridization techniques for the characterization of the human content of hybrid cell lines. PMID- 9330643 TI - Generation of a contig comprising YACs and BACs within chromosome region 1p13.1. AB - Chromosome region 1p13 is known to show loss of heterozygosity (LOH) in a number of human tumor types, including breast. We have generated a contig comprising YACs and BACs spanning part of 1p13.1 which includes the smallest region of overlapping loss identified in our earlier studies. The contig is anchored to the genetic map by a number of microsatellite markers, and by the use of CEPH YACs. We have excluded a number of candidate genes from this region, and we have oriented the contig with respect to the centromere and a number of other genes and markers on 1p13. This resource will be valuable in mapping the target for LOH in breast and other tumors, and may also be useful for the genetic analysis of other genes or diseases known to map to this region. PMID- 9330645 TI - [S(+)-ketamine bases and clinical application. Symposium as part of the German Anesthesia Congress. Hamburg, 23 April 1997]. PMID- 9330646 TI - [Olanzapin: a new perspective in the management of schizophrenia. X. Congress of the World Psychiatric Association, Madrid, August 1996]. PMID- 9330647 TI - [Anxiety and depression. Highlights from the 149th Session of the American Psychiatric Association, New York 4-9 May 1996]. PMID- 9330648 TI - [Parkinson disease--a movement disorder]. PMID- 9330649 TI - [Alzheimer dementia. Purpose of primary importance: improving the quality of life for patient and family. American Medical Association]. PMID- 9330650 TI - [Panic and depression: early diagnosis of risk patients, early therapy with SSRI (Selective Serotonin Reabsorption Inhibition)]. PMID- 9330651 TI - [Arrhythmia as risk factor]. PMID- 9330652 TI - [Hypertension in metabolic syndrome and diabetes mellitus]. PMID- 9330654 TI - [Atherothrombosis and thrombocyte function inhibitors]. PMID- 9330653 TI - [40 years of Metformin in the treatment of diabetes mellitus]. PMID- 9330655 TI - [Prevention of coronary heart disease with ACE-inhibitors]. PMID- 9330656 TI - [Annual convention of the Society of Ophthalmologists of Bavaria. Wurzburg, 19-20 July 1996]. PMID- 9330657 TI - [The Munich Glaucoma Symposium. Glaucoma: new aspects of research and therapy]. PMID- 9330658 TI - [Xalatan--a new way to treat open angle glaucoma and intraocular hypertension. Physiologic enhancement of the flow of anterior chamber fluid]. PMID- 9330659 TI - Bioproduction of indoleacetic acid by a Rhizobium sp. from the root nodules of Desmodium gangeticum DC. AB - The Rhizobium sp. isolated from the root nodules of Desmodium gangeticum DC. produced a high amount of indole acetic acid (IAA) from tryptophan in culture. For maximum IAA production, the bacteria preferred L-isomer over DL- or D-isomer of tryptophan. The production of IAA could be increased up to 37% over yeast extract ribose medium by supplementing the medium with ZnSO4 (0.1 microgram/ml), asparagine (0.1%) and nicotinic acid (0.1 microgram/ml). The possible relationship between the rhizobial IAA production and legume-rhizobia symbiosis is discussed. PMID- 9330660 TI - Biostatic and biocidal activities of water-thinnable polyesteramide compositions containing 8-hydroxyquinoline. AB - Various water soluble polyesteramide compositions are prepared by solvent technique containing a stoichiometric amount of 8-hydroxyquinoline as preservative against microbiological attack. Mechanism of action of 8 hydroxyquinoline against Pseudomonas aeruginosa ATCC 10145 is discussed and the studies showed promising results as biostatic and biocidal coatings. Preparation of water soluble polyesteramides via solvent process by using a new technique is another aspect taken into consideration. PMID- 9330661 TI - The serological signs of the Epstein-Barr virus (EBV) activity in the elderly. AB - The authors determined the IgM and IgG anti-Epstein-Barr virus (EBV)-VCA and anti EBNA levels in single serum samples of 1882 patients with diagnose except infectious mononucleosis. The serological results were divided into four groups according to different serological patterns: the seronegative group, the IgM anti VCA positive group (supposedly having actual EBV infection), the IgG (only) anti VCA group (probably recent EBV infection or not sufficiently functioning cellular immunity) and IgG anti-VCA antibodies together with anti-EBNA (past EBV infection). The presence of heterophil antibodies was detected too. The incidence rates of the serological patterns were classified according to different age groups. It has been observed that the EBV seropositivity rapidly increases with age and over 60 years of age it is over 96%. At the same time the incidence rate of the IgM anti-VCA has gradually increased with age and from 3.66% in the age group of 0-10 years reached 11.98% in patients over 60 years of age. Significantly higher titres of IgG anti-VCA were found in the elderly group of patients as well. The distribution of heterophil antibody positive samples failed to show any significant distribution. PMID- 9330662 TI - Serotyping and virulence factors of Pseudomonas aeruginosa clinical isolates. AB - Pseudomonas aeruginosa is one of the major opportunistic human pathogens very often responsible for severe infections mainly in immunocompromised hosts. These bacteria may cause the broad spectrum of infections which are associated with urinary, respiratory and gastrointestinal tract, burn wounds, eyes as well as other sites. P. aeruginosa also significantly contributes to high morbidity and mortality rates of nosocomial infections. This microorganism produces a number of exoproducts which have been implicated in the pathogenesis of pseudomonas infections. Among them, the activity of elastase, proteinase and alginate were well established. The aim of the present study was to determine the distribution of P. aeruginosa strains (isolated from urinary and respiratory tract infections) in O-serotypes and to evaluate some of their characteristics (elastase, proteinase and alginate). PMID- 9330663 TI - Contribution to the study of surface microflora on Cantal cheese. AB - Bacteria and mould were isolated from microflora of Cantal cheese rind. They were identified and the influence of pH, temperature and water activity on growth rate were studied. An equation for growth rate mu(h-1) relative to temperature is described according to a mathematical model. PMID- 9330664 TI - Changes in water activity and some microbial groups during storage of pet feed. AB - The effect of exposure of different pet feed samples to the typical temperature and moisture conditions of many months of the year (9 degrees C and 80% RH) on water activity (aw), pH and growth of various microbial groups (mesophilic aerobic bacteria, fungi and yeasts) was studied. The storage conditions were found to scarcely influence the samples pH, with no significant changes relative to the initial values (the standard deviation was 0.04 and the coefficient of variation 0.85%). By effect of exposure to the temperature and moisture conditions tested, the initially stable feeds reached moisture contents close to 20% and aw values near those of equilibrium with the ambient relative humidity during storage. Microbial counts increased with increasing aw, with final means of 8.44, 9.16 and 8.71 log CFU/g for mesophilic aerobic bacteria, fungi and yeasts, respectively. The risk of mycotoxin production at these aw values is discussed. Careful control of moisture is recommended in order to preserve the quality of pet feed throughout its scheduled shelf life. PMID- 9330666 TI - Inhibitory effect of pentoxifylline on HLA-DR expression and glycosaminoglycan synthesis of retrobulbar fibroblasts induced by interferon gamma. AB - Glycosaminoglycan (GAG) accumulation produced by retroocular fibroblasts (REF) has been observed in patients with thyroid-associated ophthalmopathy (TAO). Various cytokines are able to express HLA-DR molecules and stimulate the REF to proliferate GAG and free oxygen radicals. Pentoxifylline (Ptx) is known to have complex immunomodulatory effects on production of cytokines including interferon gamma (IFN-gamma). Ptx has been assumed to inhibit the cytokine-induced production of GAG and HLA-DR expression. We wished to determine whether Ptx has an effect on the IFN-gamma induced HLA-DR expression and influences the spontaneous and cytokine-induced GAG synthesis of REF. REF derived from extraocular muscles of healthy subjects were cultured without and with IFN-gamma. The effect of Ptx on expression of HLA-DR molecules and the production of GAG by REF was determined. Glycosaminoglycan was measured by incorporation of (3H)glycosamine into GAG. HLA-DR expression was analysed by fluorescence activated cell sorter. IFN-gamma (50, 100 and 500 U/ml) induced an increase in expression of HLA-DR molecules of REF. Ptx was proved not to be toxic for cultured cells. This drug was able to dose-dependently inhibit HLA-DR expression of REF. Both spontaneous and IFN-gamma-induced GAG synthesis of REF was inhibited by Ptx (100, 500 and 1000 mg/l, respectively). Due to in vitro inhibitory effects, Ptx is potentially able to modify the antigen presentation and the GAG synthesis by REF and it might be a useful therapeutical drug in the treatment of TAO. PMID- 9330665 TI - In vitro antimicrobial properties of azidothymidine (AZT). AB - In addition to the activity against a number of retroviruses, azidothymidine (AZT) has antibacterial activity against many bacteria. The effect of AZT on 224 bacterial species, including 25 strains of Salmonella spp. isolated from HIV positive patients, was tested. AZT had no activity against all the strains of tested Gram-positive bacteria and Pseudomonas species (MIC > 128 micrograms/ml), whereas a different activity against Enterobacteriaceae (MIC range, 128 to 0.06 micrograms/ml) was found. In particular 76% of Salmonella spp. isolated from HIV positive patients showed MICs > 1 microgram/ml, whereas similar MICs value were found in 50% of the Salmonella strains isolated from HIV-negative subjects. In addition, strains of Salmonella isolated from stools were more resistant to AZT when compared to strains isolated from blood even if this difference was not statistically significant. No correlation was found between length of therapy and Salmonella resistance to AZT in HIV-positive patients and a low incidence of Salmonella relapses in subjects treated with AZT was observed. The possibility that AZT may have an ancillary benefit in controlling some bacterial infections in AIDS patients is discussed. PMID- 9330667 TI - Shake culture studies for the production of amylases by Thermomyces lanuginosus. AB - The paper describes shake flask culture optimization studies for the production of amylases by Thermomyces lanuginosus. The culture was found to produce maximally alpha-amylase and glucoamylase (18.4 & 11.2 units/ml), respectively when the medium contained rice flour (2% w/v) as carbon and corn steep liquor as nitrogen source with medium pH adjusted to 5.5 and incubated for 72 h under shaking conditions (150 rpm) at 50 degrees C. PMID- 9330668 TI - Studies on cellulases of Aspergillus nidulans mutants. AB - Mutants were characterized as to their morphology, auxotrophy and cellulase production. No apparent correlation was evident among these characters. The degree of repression or enhancement of the three components of the cellulase enzyme in the mutants varied considerably, indicating their separate genetic control. Considerable variations in the enzyme profiles of the mutants were observed as compared to the wild type. Enzyme production was also found to be related to development. PMID- 9330669 TI - Acupuncture normalizes dysfunction of hypothalamic-pituitary-ovarian axis. AB - This article summarizes the studies of the mechanism of electroacupuncture (EA) in the regulation of the abnormal function of hypothalamic-pituitary-ovarian axis (HPOA) in our laboratory. Clinical observation showed that EA with the effective acupoints could cure some anovulatory patients in a highly effective rate and the experimental results suggested that EA might regulate the dysfunction of HPOA in several ways, which means EA could influence some gene expression of brain, thereby, normalizing secretion of some hormones, such as GnRH, LH and E2. The effects of EA might possess a relative specificity on acupoints. PMID- 9330670 TI - Increased pain threshold following electroacupuncture: analgesia is induced mainly in meridian acupuncture points. AB - Pain thresholds were determined before and after electroacupuncture of the dorsal aspect acupuncture points (AP) of the hand and non acupuncture points (NAP) located 15 mm from the traditional acupuncture points to assess changes in pain threshold thus provoked. METHODS: In eight volunteers the pain threshold of specific points was determined before and after acupuncture in the Hegu point (L.I. 4), at the back of the hand. A pressure dolorimeter was used to evaluate pain threshold at the Yangxi (L.I. 5) and Quchi (L.I. 11) points and at sites 15 mm from them. The effects on pain threshold were also measured at Yingxiang (L.I. 20) on both sides. RESULTS: Before electrostimulation there were no significant differences among the pain thresholds in both AP and NAP. After electrostimulation of the Yangxi point, pain threshold raised from 5.20 kg/sq.cm to 9.20 kg/sq.cm (p < 0.01); acupuncture at Quchi caused the threshold to increase from 5.36 kg/sq.cm to 9.20 kg/sq.cm (p < 0.01) and Yingxiang stimulation changed threshold from 2.63 kg/sq.cm to 3.83 kg/sq.cm (p < 0.051) at the point on the same side and from 2.26 kg/sq.cm to 3.90 kg/sq.cm (p < 0.05) in the opposite side. Before electroacupuncture the pain thresholds at all the tested sites were not statistically different (p > 0.1). After electrostimulation the pain threshold increased 77% at L.I. 5 but went up just 9% and 6% 15 mm from L.I. 5 (p < 0.01); threshold increased by 70% at L.I. 11 but only by 6% and 7% (p < 0.01) 15 mm from L.I. 11. CONCLUSIONS: The pain threshold increased significantly in all tested sites after electroacupuncture but the analgesic effect was predominant in those points lying along the acupuncture meridians. PMID- 9330671 TI - Effects of electroacupuncture on extracellular contents of amino acid neurotransmitters in rat striatum following transient focal cerebral ischemia. AB - In this study, we investigated the effects of electroacupuncture (EA) on extracellular levels of amino acid neurotransmitters (glutamate, aspartate, and taurine) in striatum and cerebral infarction dimensions in rats subjected to transient focal cerebral ischemia induced by 2 hours of middle cerebral artery (MCA) occlusion. EA (15 HZ, 6 mA), delivered to points of "Fengfu" (Du. 16) and "Jinsuo" (Du. 8), remarkably reduced the cerebral infarction volume. EA significantly decreased the ischemia-induced increase of extracellular aspartate level, while substantially enhancing the elevation of taurine induced by ischemia. These results indicated that the neuroprotective effect of EA against cerebral ischemia may be related to a bidirectional regulation of extracellular excitatory and inhibitory amino acid levels. PMID- 9330672 TI - A possible physical basis for the healing touch (biotherapy) evaluated by high voltage electrophotography. AB - We performed a series of experiments to examine the possibility that a theoretically proposed and indirectly empirically confirmed form of electromagnetic field emission from living beings appears to modify physical characteristics of water. We pursued three types of experiments. In the first one, we tried to examine whether and in what way water exposed to growing and dying spruce seedlings through a quartz test tube (therefore with no chemical contact), influences the germination of seeds and the growth of seedlings of the same species. The second type focused on the issue of whether and in what way distilled water, equally exposed to growing and dying spruce seedlings as well as to different ontogenetic phases of mealworm beetle, can be modified and this modification later on reproduced through a specially developed method of electrophotography. The third type of experiments presented here attempts to find out whether an emission from human hands can non-chemically modify the physical characteristics of distilled water. Their statistical analysis revealed the existence of two different groups of people: those capable of imprinting some form of highly reproducible radiation into water and those at most capable of imprinting only some sort of highly variable radiation. In the future this line of research could provide a scientifically based testing of the actual capabilities of the so-called biotherapists to perform this kind of unconventional healing. The present experiments also represent further indirect evidence for a form of electromagnetic emission from living beings and that such emission alters water in an as yet unknown way. PMID- 9330673 TI - Membrane metalloendopeptidases in immune function and disease. PMID- 9330674 TI - Structural studies of aminopeptidase P. A novel cellular peptidase. PMID- 9330675 TI - Aminopeptidase P. A cell-surface antigen of endothelial and lymphoid cells. PMID- 9330676 TI - Human lymphocyte X-prolyl aminopeptidase (aminopeptidase P)-like protein. A new member of the proline peptidase family? PMID- 9330677 TI - Specific inhibitors of aminopeptidase P. Peptides and pseudopeptides of 2-hydroxy 3-amino acids. PMID- 9330678 TI - Gamma-glutamyl transpeptidase, a blood-brain barrier associated membrane protein. Splitting peptides to transport amino acids. PMID- 9330679 TI - Structure and expression of aminopeptidase N. PMID- 9330680 TI - Activation-dependent induction of T cell alanyl aminopeptidase and its possible involvement in T cell growth. PMID- 9330681 TI - Antisense-mediated inhibition of aminopeptidase N (CD13) markedly decreases growth rates of hematopoietic tumour cells. PMID- 9330682 TI - Co-incubation of lymphocytes with fibroblast-like synoviocytes and other cell types can induce lymphocytic surface expression of aminopeptidase N/CD13. PMID- 9330683 TI - Two transfected endothelial cell lines expressing high levels of membrane bound or soluble aminopeptidase N. PMID- 9330684 TI - Aminopeptidase N-mediated signal transduction and inhibition of proliferation of human myeloid cells. PMID- 9330685 TI - Regulation of thymic development by neprilysin inhibition. AB - Development of T lymphocyte is regulated by both thymocyte-stromal cell interactions and production of soluble factors such as cytokines, peptides and hormones. The local concentration of active biological peptides is regulated by a specialized family of enzymes expressed at the cell surface, the ectopeptidases. We found that treatment of fetal thymic organ cultures (FTOC) with the specific CD10 (endopeptidase 24.11) inhibitor, (N-(3-[(hydroaxyamino)carbonyl]-2 benzylidene-1-oxopropyl]-N-glyci ne), RB25 results in a marked delay in thymocyte differentiation. RB25 causes a significant decrease in the number of DP (CD4+CD8+) cells in favor of the TN (TcR alpha beta-CD4-CD8-) population. RB25 also blocks T lymphocyte differentiation in FTOC when preinjected into pregnant mice. Finally, RB25 was found to essentially affect the CD44+CD25- and CD44-CD25- thymocytes in "in vitro" and "in vivo" experiments after 2 days FTOC. Thus, a selective and stable endopeptidase 24.11 inhibitor impairs T cell development, an observation in agreement with the involvement of the CD10 antigen in early T cell development. PMID- 9330686 TI - Proteases of isolated pancreatic acinar cells after caerulein hyperstimulation. PMID- 9330687 TI - Structure of CD26 (dipeptidyl peptidase IV) and function in human T cell activation. PMID- 9330688 TI - Molecular associations required for signalling via dipeptidyl peptidase IV (CD26). PMID- 9330689 TI - CD26/dipeptidyl peptidase IV in lymphocyte growth regulation. AB - DP IV/CD26 is involved in regulation of DNA synthesis and proliferation as well as production of cytokines of hematopoietic cells under various conditions. Inhibition of DNA synthesis in T lymphocytes, B lymphocytes, NK cells and myelomonocytic cells as well as of the production of IL-2, IL-6 TNF alpha, IL-1, IL-10, IL-12, IL-13, IFN-gamma, GM-CSF are not due to apoptosis of these cells. DP IV/CD26 inhibitors induce TGF-beta 1 mRNA synthesis and latent protein release demonstrating a crucial role of TGF-beta 1 in mediating CD26 function. X-X-Pro peptides as HIV-Tat protein strongly inhibit DP IV enzymatic activity and suppress DNA synthesis. This group of peptides may represent a class of natural DP IV/CD26 ligands and effectors, respectively. Hyperphosphorylation of p56lck as well as protein tyrosine phosphorylation of a number of proteins in T lymphocytes can be modulated by DP IV inhibitors. These data suggest that enzymatic activity or, at least in part, the active site of DP IV are both essential for its regulatory function in lymphocytes. Further work is required to determine the natural ligands, i.e. substrates and effectors, which are play the central role in DP IV/CD26 action in T cell growth and to understand the molecular mechanism of the early steps of this fundamental process. PMID- 9330690 TI - CD26 is involved in regulation of cytokine production in natural killer cells. PMID- 9330691 TI - The effect of anti-CD26 antibodies on DNA synthesis and cytokine production (IL 2, IL-10 and IFN-gamma) depends on enzymatic activity of DP IV/CD26. PMID- 9330692 TI - New fluorogenic dipeptidyl peptidase IV/CD26 substrates and inhibitors. PMID- 9330693 TI - Molecular analyses of human and rat dipeptidyl peptidase IV. PMID- 9330694 TI - A molecular model of the active site of dipeptidyl peptidase IV. Explanation of the substrate specificity and interaction with inhibitors. PMID- 9330695 TI - The level of CD26 determines the rate of HIV entry in a CD4+ T-cell line. AB - We have reported that CD26 could serve as a cofactor of CD4 in HIV entry. Recently, more evidence has been provided for the implication of CD26 in HIV entry, replication and cytopathic effect. Along with, we have demonstrated that the level of CD26 may determine the rate of HIV-envelope induced-apoptosis. The role of CD26 in HIV entry was further investigated using CEM T-cell line. Clones were established by transfection, expressing different levels of CD26. Entry, infection and cytopathic effect were monitored in several independent clones, and were found to be delayed in clones CD26-Low and CD26-SuperHigh compared to clones CD26-High. The delay was most significant in clones CD26-AntiSense, without any apparent cytopathic effect. These results demonstrate that relatively enhanced levels of CD26 contribute to an increased virus infection. Furthermore, they illustrate that CD26-SuperHigh clones manifest a phenotype similar to CD26-Low clones. This point out the critical role of CD26 in the rate of HIV entry and its cytopathic effect, two events which are initiated by the interaction of HIV envelope glycoproteins with cell-surface CD4. PMID- 9330696 TI - HIV-1 envelope gp120 and viral particles block adenosine deaminase binding to human CD26. AB - CD26, known to be the adenosine deaminase (ADA) binding protein, has been implicated in HIV infection. In human B and T cell lines we show that, irrespective of CD4 expression, 125I-labeled ADA binding to CD26 is inhibited by recombinant soluble HIV-1 envelope glycoprotein gp120 and by HIV-1 infectious particles. Overlapping synthetic peptides covering the entire gp120 sequence were tested to map the region in gp120 responsible for ADA binding inhibition. Only peptides in the C3 region significantly inhibited the binding of ADA to CD26. These results indicate that a specific function of gp120 is the inhibition of ADA binding to CD26 in both CD4+ and CD4- cells. Since the interaction ecto-ADA/CD26 is required for the activation of T cells, it remains possible that HIV particle mediated blockade of ecto-ADA/CD26 interaction may have significant consequences in the pathogenesis of AIDS disease. PMID- 9330697 TI - Further characterization of DPP IV-beta, a novel cell surface expressed protein with dipeptidyl peptidase activity. AB - By using a CD26 negative human lymphoblastoid cell line (C8166), here we describe the characterization of a cell-surface protein which manifests CD26-like dipeptidyl peptidase IV (DPP IV) activity. This protein, referred to as DPP IV beta, shows a higher KM value for Gly-Pro-pNA than CD26 (0.31 mM compared to 0.11 mM, respectively). In addition, DPP IV-beta was found not to bind 125I-labeled adenosine deaminase (a property of human CD26). Gel filtration experiments using extracts from C8166 and MOLT4 (a CD26 positive human T cell line) cells, revealed that the apparent molecular mass of DPP IV-beta is 82 kDa, whereas that of CD26 is 110 kDa. In order to conveniently differentiate both activities, a new family of inhibitors, that selectively blocks peptidase activity associated to CD26, has been developed. PMID- 9330698 TI - Expression of dipeptidylpeptidase IV (DPP IV/CD26) activity on human myeloid and B lineage cells, and cell growth suppression by the inhibition of DPP IV activity. PMID- 9330699 TI - CD26 as a positive regulator of HIV envelope-glycoprotein induced apoptosis in CD4+ T cells. AB - The membrane-expressed HIV-1 envelope glycoprotein complex, gp120/gp41, has been shown to be responsible for the initiation of cell killing by apoptosis in CD4+ T cells. By using two experimental approaches we demonstrate that CD26, independent of its DPP IV activity, appears to be implicated in this function of the gp120/gp41 complex to initiate apoptosis. PMID- 9330700 TI - Comparative study of CD26 as a Th1-like and CD30 as a potential Th2-like operational marker in leprosy. AB - In the last years we have been able to establish CD26 as an operational marker for a human Th1-like reaction in various granulomatous diseases. Recently, CD30 was described as a marker for a Th2-type reaction, where CD30 is preferentially expressed and its soluble form released by human T cell clones producing Th2-type cytokines. To evaluate the possibility of CD30 as an eventual operational marker for a human Th2-like reaction in vivo, we performed immunohistological stainings on frozen sections of skin biopsies from patients with lepromatous and tuberculoid leprosy. A maximum of three to four CD30-positive cells was found per section, and there was no difference in the accumulation of CD30-positive cells between the tuberculoid and the lepromatous form of leprosy. With respect to CD26 positive cells, a high number was found in tuberculoid leprosy in contrast to a greatly reduced expression of CD26 in lepromatous leprosy. We conclude that, while CD26 was confirmed as an operational marker for a Th1-like reaction in leprosy, CD30 does not represent an operational Th2 marker in this disease. PMID- 9330701 TI - Regulation of neutrophil activation by proteolytic processing of platelet-derived alpha-chemokines. PMID- 9330702 TI - Selective proteolytical cleavage of the ligand-binding chains of the IL-2 receptor and IL-6-receptor by neutrophil-derived proteases. PMID- 9330703 TI - In vitro effects of gamma-glutamyltranspeptidase inhibitor acivicin on human myeloid and B lineage cells. PMID- 9330704 TI - Expression of several matrix metalloproteinase genes in human monocytic cells. AB - Matrix metalloproteinases (MMP) are a family of structurally related endopeptidases that resorb macromolecules of the extracellular matrix (ECM). They are involved in normal tissue remodeling and wound repair as well as in pathological processes such as the irreversible destruction of joints observed in rheumatoid arthritis (RA). In addition, MMP catalyze the cleavage of the transmembrane form of tumor necrosis factor (TNF). Since cells of the monocyte lineage are major producers of TNF in the rheumatoid synovium we analysed the expression of MMP genes in these cells. To examine the transcriptional activity of MMP genes in undifferentiated monocytic cell lines (MonoMac6, U937) and in nature human monocytes isolated from peripheral blood, we developed an assay that is based on reverse transcription (RT) followed by a polymerase chain reaction (PCR). This screening procedure demonstrates that several MMP genes are transcriptionally active in the cells tested after exposure to a variety of stimuli such as phorbol ester, lipopolysaccharide (LPS) and staphylococcal enterotoxin B (SEB). The data were confirmed by quantitative Northern blot analysis. In conclusion, cells of the monocyte lineage produce high mRNA levels of at least six members of the MMP gene family that could participate in joint destruction by resorption of the ECM and secretion of TNF. PMID- 9330705 TI - Lysosomal cysteine peptidases and malignant tumours. PMID- 9330706 TI - Expression of cysteine protease inhibitors stefin A, stefin B, and cystatin C in human lung tumor tissue. AB - In human lung tumor tissue specimen (n = 73) concentrations of stefins A and B were found to be increased 2.0-fold (p < 0.01) and 1.3-fold (p < 0.01), respectively, as compared to matched normal tissue. Stefin A and B concentrations were higher in primary tumors than in secondary tumors, i.e. metastases from other organs to the lung (p < 0.01; p < 0.05, respectively). Cystatin C concentrations were rather low and did not differ between tumor and normal tissue. Both concentrations of stefins did not correlate with TNM stages. Stefin A was higher in squamous cell carcinoma than in adenocarcinoma (p < 0.01), while stefin B did not show such a difference. At investigation of a relationship between survival probability of patients with primary tumors it was found that increased stefin B concentrations and total cysteine-protease-inhibitory activities but not stefin A concentrations were positively correlated with survival probability. It is concluded that stefins A and B are major contributors to the cysteine protease inhibitory activity in primary lung tumors. Stefin B proved to be a prognostic factor, especially in squamous cell carcinoma. PMID- 9330707 TI - Contribution of the proteasome to the alpha-secretase pathway in Alzheimer's disease. PMID- 9330708 TI - Dipeptidyl peptidase IV (CD26) and Alzheimer amyloid protein precursor (APP) in polymyositis. PMID- 9330709 TI - The HIV protease and therapies for AIDS. AB - New, potent therapies for HIV disease are available, based on synthetic inhibitors of the viral protease, an essential viral enzyme. The results in clinical trials have been impressive with most treated individuals benefiting in terms of reduced quantity of detectable virus, enhanced numbers of CD4 lymphocytes and improvements in quality and duration of life. However, there are some remaining negatives associated with the new drugs, including high cost, side effects and appearance of drug-resistant strains of HIV. Problems and future prospects for use of protease inhibitors and alternate approaches in AIDS are discussed. PMID- 9330710 TI - Leukodiapedesis, function, and physiological role of leucocyte matrix metalloproteinases. PMID- 9330711 TI - Matrix metalloproteinases in experimental autoimmune encephalomyelitis. PMID- 9330712 TI - Interaction of transforming growth factor beta (TGF beta) with proteinase 3. AB - TGF beta is a multifunctional cytokine modulating onset and course of autoimmune diseases as shown in experimental models. Aim of this study was to investigate possible interactions of TGF beta with lysosomal enzymes identified as ANCA autoantigens (e.g. proteinase 3, PR3). This included TGF beta effects on the translocation the lysosomal enzymes to the cell surface of polymorphonuclear cells (PMN), and the presumabe activation of non bioactive, latent TGF beta by these enzymes. Flow cytometry analysis showed TGF beta 1 to be a potent translocation factor for PR3 comparable with other neutrophil activating factors such as interleukin 8 (IL8). The PR3 membrane expression on primed PMN increased by up to 51% after incubation with TGF beta 1. PR3 itself was revealed as a potent activator of latent TGF beta, thus mediating bioeffects of this cytokine. Patients with various types of systemic vasculitis (SV) showed marked TGF beta overexpression correlating with disease. Mean TGF beta 1 plasma levels in the ANCA associated vasculitis (AAV) patients ranged from 8.9 (Wegeners granulomatosis, WG) to 13.3 ng/ml (Churg-Strauss syndrome, CSS)(control: 4.2 ng/ml, p < 0.01) while TGF beta 2 levels were not elevated. Our findings, together with other features of TGF beta's such as induction of angiogenesis and its strong chemotactic capacity, indicate that TGF beta might serve as a proinflammatory factor in SV, especially in AAV. PMID- 9330713 TI - Liver cysteine proteinases in macrophage depression induced by gadolinium chloride. AB - Liver lysosomal enzymes during macrophage depression (gadolinium chloride, 7 mg/kg, intravenously) and macrophage stimulation (zymosan, 100 mg/kg, intravenously) have been studied. It was shown that gadolinium chloride treatment of rats reduced the rate of carbon particles phagocytosis at 24 and 48 h after the single administration. Decreased endocytic capacity of Kupffer cells was confirmed also by electron microscopy. Gadolinium chloride induced labilization of liver lysosomes (increased free activity of cathepsins B and L); there was no changes of specific activity of liver cysteine proteinases (24 h). Gadolinium chloride prevented death of rats after administration of non-sonicated particular zymosan particles, resulting to 70% survival, compare with the 17% survival in group with zymosan alone. We can summarize that macrophages depression by gadolinium chloride abolish symptoms of inflammation in zymosan-model, influencing on cysteine proteinases of Kupffer cells. PMID- 9330714 TI - Ca(2+)-signaling in cardiac myocytes: evidence from evolutionary and transgenic models. AB - Cardiac contraction is regulated by a number of Ca(2+)-mediated processes. Here we consider the effects of modification imposed on the Ca(2+)-signalling mechanism by evolutionary developments and transgenic manipulations. Ca(2+) signalling appears to be mediated via influx of Ca2+ through the DHP receptor in preference to the Na(+)-Ca2+ exchange protein, and activates the ryanodine receptor and the Ca2+ release from the SR. Here we report on functional consequences of overexpression of the Na(+)-Ca2+ exchanger and calsequestrin. The data does not support a physiological role for the Na(+)-Ca2+ exchanger in signalling Ca2+ release, but can serve to modify ionic currents which determine the duration of the action potential. PMID- 9330715 TI - Diastolic viscoelastic properties of rat cardiac muscle; involvement of Ca2+. AB - Diastolic cardiac sarcomere stiffness, sarcomere length changes, and calcium concentration [Ca2+]i were investigated in 18 trabeculae, dissected from the right ventricle of rat heart. [Ca2+]i declined following a mono-exponential diastolic time course with a time constant of 210-350 ms. During diastole, ([Ca2+]o = 1 mM); sarcomere length (SL) increases (amplitude: 5-65 nm; time constant: 600 ms). Eighty percent of muscles showed discrete spontaneous motion of sarcomeres near the end of diastole; this phenomenon occurred earlier at higher [Ca2+]o. The stiffness modulus of the sarcomere (MOD) increased by 30% during diastole (n = 158; p < 0.05), while the phase difference, phi, between force and SL decreased by 13% (n = 158; p < 0.05). The increase of MOD and the decrease of phi reversed when spontaneous activation occurred. These results show that the mechanical diastolic properties of the cardiac sarcomere are time dependent. The time dependence of the diastolic properties can be faithfully reproduced by a simple linear four element viscoelastic model. The diastolic changes of MOD and of phi could be reproduced by assuming an exponential change of the elastic and viscous coefficients of the model over time with a time constant similar to the time constant of change of [Ca2+]i. We suggest that the simplest combination of structural counterparts of the model in the sarcomere consists of titin bound to both actin and myosin in the myofibril, while the sarcomere is in parallel with another purely elastic element. We propose that the Ca(2+)-dependence of diastolic stiffness might be the result of an inverse relation between [Ca2+]i and the affinity of titin for actin. PMID- 9330716 TI - The beta subunit, Kv beta 1.2, acts as a rapid open channel blocker of NH2 terminal deleted Kv1.4 alpha-subunits. AB - A recently discovered class of ancillary subunits has been shown to modify the inactivation properties of alpha-subunits belonging to the Kv1 family of potassium channels. One of these subunits, Kv beta 1.2, modifies intrinsic alpha subunit C-type inactivation. N-type inactivation and open channel block have been proposed to increase the rate of development of C-type inactivation. We demonstrate here that Kv beta 1.2 has kinetic properties which are consistent with rapid open channel block. PMID- 9330717 TI - Signal transduction in ischemic preconditioning. AB - Ischemic preconditioning is a phenomenon in which exposure of the heart to a brief period of ischemia causes it to quickly adapt itself to become resistant to infarction from a subsequent ischemic insult. The mechanism is not fully understood but, at least in the rabbit, it is known to be triggered by occupation of adenosine receptors, opioid receptors, bradykinin receptors and the generation of free radicals during the preconditioning ischemia. All of these are thought to converge on and activate protein kinase C (PKC), which in turn activates a tyrosine kinase. This kinase cascade eventually terminates on some unknown effector, possibly a potassium channel or a cytoskeletal protein, which makes the cells resistant to infarction. If this process can be understood, it should be possible to devise a method for conferring this protection to patients with acute myocardial infarction. PMID- 9330718 TI - Ca2+ sparks within 200 nm of the sarcolemma of rat ventricular cells: evidence from total internal reflection fluorescence microscopy. AB - Total internal reflection fluorescence microscopy (TIRFM) was used to measure local calcium releases in resting cardiac myocytes stained with fluo-3AM. The measured fluorescence originated from regions where cells were close to, and develop adhesions to, a totally reflecting glass surface. The excitation of the fluorescent Ca2+ indicator dye by the exponentially attenuated evanescent wave penetrated approximately 200 nm into the fluid phase. In rat ventricular cells, Ca2+ waves and Ca2+ sparks were observed within the adhesions. Ca2+ sparks recorded with TIRFM compared favorably to sparks recorded under similar conditions with confocal microscopy. Computer simulation supported this assessment. It is concluded that TIRFM can provide an economical, flexible tool for detailed measurement of Ca(2+)-transients in the subsarcolemmal space of live cells. PMID- 9330720 TI - Molecular control of myocardial mechanics and energetics: the chemo-mechanical conversion. AB - Energy consumption in the cardiac muscle is characterized by two basic phenomena: 1) The well known linear relationship between energy consumption by the sarcomere and the mechanical energy it generates, and 2) the ability to modulate the generated mechanical energy and energy consumption to the various loading conditions, as is manifested by the Frank-Starling Law and the Fenn effect. These basic phenomena are analyzed here based on coupling calcium kinetics with crossbridge (Xb) cycling. Our previous studies established the existence of two feedback mechanism: 1) a positive feedback mechanism, the cooperativity, whereby the affinity of the troponin for calcium, and hence Xb and actomyosin-ATPase recruitment, depends on the number of force generating Xbs, and 2) a mechanical feedback, whereby the filaments shortening velocity, or the Xb strain rate, determines the rate of Xb turnover from the strong to the weak conformation. The cooperativity mechanism determines the force-length relationship (FLR) and the related Frank-Starling Law. It also provides the basis for the regulation of energy consumption and the ability of the muscle to adapt its energy consumption to the loading conditions. The mechanical feedback regulates the shortening velocity and provides the analytical solution for the experimentally derived Hill's equation for the force-velocity relationship (FVR). The mechanical feedback regulates the generated power and provides the linear relationship between energy consumption and the generated mechanical energy, i.e., the external work done and the liberated heat. Thus, the two feedback mechanisms that regulate sarcomere dynamics, and determine the FLR and FVR, also regulate the energy consumption and the mechanical energy generated by the muscle. PMID- 9330719 TI - Uncoupling of G-protein coupled receptors in vivo: insights from transgenic mice. AB - Heart failure is a problem of increasing importance in medicine. An important characteristic of heart failure is reduced agonist-stimulated adenylyl cyclase activity (receptor desensitization) due to both diminished receptor number (receptor down regulation) and impaired receptor function (receptor uncoupling). These changes in the beta-adrenergic receptor (beta-AR) system, may in part account for some of the abnormalities of contractile function in this disease. Myocardial contraction is closely regulated by G-protein coupled beta-adrenergic receptors through the action of the second messenger cAMP. The beta-AR receptors themselves are regulated by a set of specific kinases, termed the G-protein coupled receptor kinases (GRKs). The study of this complex system in vivo has recently been advanced by the development of transgenic and gene targeted ("knockout") mouse models. Combining transgenic technology with sophisticated physiological measurements of cardiac hemodynamics is an extremely powerful strategy to study the regulation of myocardial contractility in the normal and failing heart. PMID- 9330721 TI - Myocardial cell energetics. AB - Energy transformation at the main energy consuming processes of the myocardium takes place with high efficiency, i.e., with relatively small differences between the free energy level provided and the free energy level required for the two coupled processes. Thus, the free energy of ATP is only moderately higher than that of various ATP dependent processes. Under energy deficiency caused by hypoxia, free energy of ATP can drop to a level that critically affects subsequent steps. Detailed evaluation of cell energetics was carried out with the following approach: Cell shortening, oxygen consumption and intracellular calcium transients of isolated rat cardiomyocytes which were investigated under the influence of inotropic interventions. Increased extracellular Ca2+ and isoproterenol reduced the economy of contraction (contraction amplitude/VO2), whereas Ca-sensitizing agent EMD 57033 did not. This seems to be a consequence of the increased costs of ion cycling under the effect of Ca2+ and isoproterenol. Our current investigation suggests that alterations of ion transport processes and crossbridge kinetics have substantial impact on myocardial energetics. PMID- 9330722 TI - Human heart failure: determinants of ventricular dysfunction. AB - Thin muscle strips were obtained from non-failing (NF) and failing (dilated cardiomyopathy (DCM)) hearts, using a new harvesting and dissection technique. The strips were used to carry out a myothermal and mechanical analysis so that contractile and excitation coupling phenomena in the NF and failing (DCM-F) preparations can be compared. Peak isometric force and rate of relaxation in DCM F were reduced 46% (p < 0.02) while time to peak tension was increased 14% (p < 0.03). Initial, tension dependent, tension independent and the rate of tension independent heat liberation were reduced 62-70% in DCM-F (p < 0.03). The crossbridge force-time integral (FTIXBr) was calculated from these measurements and was shown to increase 40% while the amount and rate of calcium cycled per beat was reduced 70%. As a result of these changes in the contractile and excitation-contraction coupling systems in DCM-F, the force-frequency relationship was significantly blunted while the power output was markedly reduced. These fundamental alterations account for the substantial ventricular dysfunction found in the dilated cardiomyopathic failing heart. PMID- 9330723 TI - How cardiac contraction affects the coronary vasculature. AB - We modeled the influence of cardiac contraction on maximally dilated coronary blood vessels, whether single or in juxtaposition, taking into account the nonlinear material properties of both the vascular wall and the myocardium. We calculated pressure-area relations of single, embedded coronary blood vessels, and used these relations to calculate diastolic and systolic coronary pressure flow relations in a model of the coronary vasculature. The model shows that the change in myocardial material properties during contraction can explain the decrease in coronary vessel area and coronary flow generally observed in experiments. The model also shows that arterioles can be protected from the compressive action of the cardiac muscle by the presence of accompanying venules, which is favorable for coronary blood flow. PMID- 9330724 TI - Dynamic interaction between myocardial contraction and coronary flow. AB - Phasic coronary flow is determined by the dynamic interaction between central hemodynamics and myocardial and ventricular mechanics. Various models, including the waterfall, intramyocardial pump and myocardial structural models, have been proposed for the coronary circulation. Concepts such as intramyocardial pressure, local elastance and others have been proposed to help explain the coronary compression by the myocardium. Yet some questions remain unresolved, and a new model has recently been proposed, linking a muscle collagen fibrous model to a physiologically based coronary model, and accounting for transport of fluids across the capillaries and lymphatic flow between the interstitial space and the venous system. One of the unique features of this model is that the intramyocardial pressure (IMP) in the interstitial space is calculated from the balance of forces and fluid transport in the system, and is therefore dependent on the coronary pressure conditions, the myocardial function and the transport properties of the system. The model predicts a wide range of experimentally observed phenomena associated with coronary compression. PMID- 9330725 TI - The relations between microvascular structure and oxygen supply. AB - The current theories of microcirculatory control require a higher command center to maintain tissue homeostasis. The theory of arteriolar vasodilation generates conflicting results by increasing the flow and the capillary hydrostatic pressures. A new theory, recently suggested to account for local tissue flow control, applies a blinking mechanism of open capillaries, whereby alternating circulatory modes increase flow to specific tissue regions, without inducing a higher control center. The mechanism is evaluated here mathematically by examining the oxygen demand, supply and local tissue concentration of oxygen. It is shown that this theory explains how periods of oxygen debts are compensated by increased flow with changes in the operation mode. The results show that there is a direct relation between the tissue microvascular complexity and the capability of the specific tissue to withstand periods of oxygen deficit. PMID- 9330726 TI - Endothelial gene regulation by laminar shear stress. AB - Endothelial cells, because of their unique localization, are constantly exposed to fluid mechanical forces derived by the flowing blood. These forces, and more specifically shear stresses; affect endothelial structure and function, both in vivo and in vitro, and are implicated as contributing factors in the development of cardiovascular diseases. We have demonstrated earlier that the shear stress selectively induces the transcription of several endothelial genes, and have defined a shear stress response element (SSRE) in the promoter of platelet derived-growth-factor B (PDGF-B), that is shared by additional endothelial shear stress responsive genes. Here we further characterize this SSRE and the nuclear factors that bind to it, and imply the possible role of the endothelium cytoskeleton in transducing shear stress, leading to the expression of PDGF B/SSRE constructs in transfected endothelial cells exposed to shear stress. We also present, yet a new shear stress response element in the Platelet Derived Growth Factor A promoter, that contains a binding site to the transcription factors egr1/sp1. These results further demonstrate the complexity of gene regulation by hemodynamic forces, and support the important part that these forces have in the physiology and pathophysiology of the vessel wall. PMID- 9330727 TI - 3D architecture of myocardial microcirculation in intact rat heart: a study with micro-CT. AB - The branching geometry of the coronary arterial tree may play a significant role in the observed spatial heterogeneity in myocardial perfusion. To provide more insight into this possibility we used a micro-CT scanner to image the intact rat heart and its opacified coronary arterial tree, for quantitative analysis of the coronary arterial architecture. Results show a consistent pattern of branching throughout the heart wall. PMID- 9330728 TI - Vascular imaging by ultrasound: 3D reconstruction of flow velocity fields for endothelial shear stress calculation. AB - A new method for quantitative reconstruction of a three dimensional (3D) velocity field from ultrasound color doppler mapping (USCDM) images is used here to calculate the shear stress distribution on the endothelial layer of an artery. Measurements of a few spatially unrestricted USCDM transverse cross sectional images of the artery, and of several echo-ultrasound B-mode images of the same area, are required for reconstructing the geometry of the vessel's endothelial surface. The calculation is based on assuming a physical model of flow, and solving the Continuity and the Navier-Stokes equations numerically for a steady flow of an incompressible Newtonian fluid at constant temperature within a non flexible tube. The correct choice of the penalty parameter in the finite element method (FEM) algorithm provides proper convergence of the reconstruction. The endothelial shear stress is calculated from the gradient of the velocity field at each point of the vessel's inner surface. PMID- 9330730 TI - Atherosclerosis studies by intracoronary ultrasound. AB - Intravascular ultrasound (IVUS) is a new technique of tomographic visualization of the coronary arteries: its lumen, wall and pathology. Three dimensional (3D) reconstruction shows the tubular structure of the arterial wall and its pathology. IVUS has many advantages over coronary angiography: it has better resolution and shows many hidden lesions. IVUS has helped uncover the underlying mechanisms of percutaneous transluminal coronary angioplasty (PTCA), restenosis, the use and value of other interventional techniques such as directional coronary atherectomy (DCA), rotational atherectomy and stent implantation, and has great value in planning complex interventional procedures. The new American Heart Association (AHA) classification of coronary atherosclerosis pathology can be demonstrated by IVUS. IVUS is sensitive for studies of atheroma regression and progression and shows the coronary artery lesions after cardiac transplantation. PMID- 9330729 TI - Compensatory enlargement, remodeling, and restenosis. AB - Coronary atherosclerosis is associated with vessel remodeling and dilatation. Quantitative analysis of arterial morphometry documents that as the cross sectional area of the plaque increases within a diseased vessel segment, the outer wall of the artery expands in an attempt to compensate for the accumulation of plaque. This focal compensatory enlargement maintains cross sectional area at stenotic sites of arteries, and the angiographic appearance of the vessel may be normal, despite marked accumulation of atherosclerotic plaque. Compensatory enlargement may develop as a response to increased shear stress caused by atherosclerotic plaque in conjunction with endothelial dependent factors, or alternatively, due to medial attenuation with loss of underlying structural support. The mechanism of arterial remodeling plays an important role in the development of arterial restenosis late after balloon dilatation or other interventional modalities. Stents prevent the remodeling process and restenosis with stents is the result of a relatively uniform neointimal tissue proliferation throughout the stent. Endovascular stents induce tissue proliferation both within the endoluminal stent surface and in the tissue layers surrounding the metallic Palmaz-Schatz stent struts. The therapeutic goals to prevent restenosis have to be oriented towards prevention of acute recoil, reduction of intimal, medial and adventitial prolific response and the phenomenon of inadequate arterial remodeling. PMID- 9330731 TI - Tissue remodeling with micro-structurally based material laws. AB - Cardiomyocytes and the extracellular collagen matrix which holds them together respond to changes in their mechanical environment by adapting their orientation, size and composition. We examine local mechanical feedback mechanisms affecting the fiber orientation, sheet orientation and passive fiber direction stiffness, using an axisymmetric finite element model of the left ventricle (LV), with material constitutive laws based on the fibrous-sheet microstructure of myocardium. PMID- 9330732 TI - Multiaxial myocardial mechanics and extracellular matrix remodeling: mechanochemical regulation of cardiac fibroblast function. AB - Substantial evidence suggests that not only does the structure of the cardiac extracellular matrix affect the mechanical properties of myocardium, but that mechanical loading affects the synthesis of the extracellular matrix. However, loading conditions in vivo are nonhomogeneous and multiaxial. An experimental approach that combines mechanics and cell biology is used to examine the mechanisms of extracellular matrix remodeling in the heart. The results indicate that differential biological responses in adult cardiac fibroblasts can be correlated with specific physical signals, such as the magnitude and two dimensional (2D) pattern of strain. Some effects of flow-function relations are discussed. PMID- 9330733 TI - In vivo assessment of regional myocardial work in normal canine hearts using 3D tagged MRI. AB - A non-invasive method for assessing regional myocardial work is presented. The method utilizes tagged magnetic resonance images (MRI) obtained from two sets of orthogonal planes to mark and reconstruct 24 small myocardial cuboids at end diastole (ED) and end-systole (ES) in the in vivo left ventricle (LV). Regional myocardial work is assessed by calculating the area enclosed by the endocardial wall tension-area (T-A) loop of each studied cuboid. The method was applied to six normal canine hearts. In addition, a global myocardial work index was obtained from the corresponding estimated pressure-volume (P-V) loops. The average work index calculated using the T-A loop was 0.242 +/- 0.088 J/100gr/beat, in agreement with the average index obtained from the P-V loop: 0.296 +/- 0.089 J/100gr/beat. The two indices correlate linearly with a correlation coefficient of 0.82. PMID- 9330734 TI - Noninvasive measurement of cardiac strain with MRI. AB - The motion sensitivity of cardiac magnetic resonance imaging (MRI) can be exploited to measure the motion patterns within the heart wall and thus to noninvasively calculate the intramyocardial strain. The resulting large data sets pose a challenge for visualization, but offer the potential of a greatly improved picture of cardiac dynamics. This may have both basic research and clinical applications. PMID- 9330735 TI - Ventricular remodeling: from bedside to molecule. AB - The multiple mechanisms that bring about the decompensation of the hypertrophic remodeled myocardium are synergistic and not fully understood. Our current hypothesis is that the increased stress on the ventricle is initially offset by compensatory myocardial hypertrophy. In many instances, however, progressive ventricular dilatation and heart failure occur as a result of maladaptive hypertrophy (abnormal myosin-actin production), programmed cell death (apoptosis) and/or changes in the interstitial vasculature and collagen composition. The molecular and genetic background to these processes includes changes in myocardial gene expression, activation of the local tissue renin-angiotensin and other neurohormonal systems, increased matrix metalloproteinase activity (including collagenase), and expression of certain components of the immune system, such as TNF-alpha. Future research will hopefully provide better methods for limiting the remodeling-ventricular dilatation process by novel pharmacotherapies, gene therapy and, possibly, surgical therapy, and determine the impact of such interventions on survival. PMID- 9330736 TI - Cardiac excitation: an interactive process of ion channels and gap junctions. AB - Theoretical simulations were performed to study the interplay between membrane ionic currents and gap-junction coupling in determining cardiac conduction. Results demonstrate that a much slower conduction velocity can be achieved with reduced gap-junction coupling than with reduced membrane excitability. Also, uniform reduction in intercellular coupling increases spatial asymmetries of excitability and, consequently, the vulnerability to unidirectional block. PMID- 9330737 TI - Modeling of internal pH, ion concentration, and bioenergetic changes during myocardial ischemia. AB - Arrhythmias are caused by the interdependent processes of change in energy metabolism and alterations in sarcolemmal ion gradients that occur during ischemia. Depletion of energy metabolites and increased proton concentrations in ischemic heart may underlie the observed phenomena of reduced contractile force and also of malignant ventricular arrhythmias that can lead to tachycardia and ventricular fibrillation. Recent advances in measuring changes in ion concentrations and metabolites during cardiac ischemia have provided a wealth of detail on the processes involved. Some of the experimental data have been used to construct a computer model that integrates cardiac energetics with electrophysiological changes. This is a novel approach to studying myocardial ischemia, and the resulting model would aid in the prediction of the effects of therapeutic interventions. PMID- 9330738 TI - Gap junctions: functional effects of molecular structure and tissue distribution. AB - Abnormal conduction is fundamental to the pathogenesis of both atrial fibrillation and ventricular tachycardia/fibrillation. Normal atrial and ventricular myocytes express different combinations of multiple gap junction channel proteins and are interconnected by gap junctions in markedly different spatial distributions. These observations suggest that the disparate anisotropic conduction properties of atrial and ventricular muscle are determined, in part, by both structural and molecular features of gap junctions. Alterations in gap junctional coupling likely contribute to conduction abnormalities underlying reentrant atrial or ventricular arrhythmias. PMID- 9330740 TI - Percutaneous multielectrode endocardial mapping and ablation of ventricular tachycardia in the swine model. AB - A basket shaped catheter carrying 64 electrodes was developed in the left ventricle (LV) of 53 pigs which had undergone induction of myocardial infarction. Pacing during sinus rhythm, or echocardiographic and hemodynamic measurements as well as pathological studies revealed no significant damage due to the basket catheter. Eighty one episodes of ventricular tachycardia (VT) were mapped and analyzed, requiring only several beats and less than 10 seconds to complete. We were able to successfully ablate ventricular tachycardias in four pigs. PMID- 9330739 TI - 3D cardiac imaging of electromechanical coupling. AB - A novel method for three dimensional (3D) electromechanical mapping of the heart is presented. The new method is based on utilizing special magnetically locatable catheters connected to a mapping and navigation system. The 3D electromechanical map of the chamber is reconstructed by sampling the location of the catheter tip throughout the cardiac cycle at a plurality of endocardial sites together with their local electrograms. The ability to spatially combine electrical and mechanical information may provide a useful tool for both research and clinical cardiology. PMID- 9330741 TI - Design and strategy for the Cardionome Project. AB - The Physiome Project has the goal of providing the quantitative description of the integrated functions of the living organism. This is too large an undertaking to be begun all at once. What needs to be developed first are large comprehensive databases containing genomic, biochemical, anatomical and physiological information that can be searched and retrieved via the Internet. A more modest and achievable goal is the Cardiome Project, whose goal is to describe the functioning heart. Since it is impractical to develop this from the genetic and molecular level, we begin it as a multicenter collaborative effort at the level of the functioning organ. The work of Hunter, Noble, and others provides a central scheme, a description of the spread of excitation and contraction through an anatomically detailed cardiac model with fiber directions. This will be augmented by the additions of regional blood flows, substrate uptake and metabolism, and energy production and utilization in serving contraction and ionic balances. Later stages will involve cellular regulation and responses to interventions. The organization of such projects is by the assembling of components whose linkages one to another are first minimized and then augmented to improve the approximation to reality. PMID- 9330742 TI - Integrative and interactive studies of the cardiac system: deciphering the cardionome. AB - The cardiac system, denoted as the Cardionome, represents one of the most exciting challenges to human ingenuity. Critical to our survival, it consists of a tantalizing array of interacting phenomena, from ionic transport, membrane channels and receptors through cellular metabolism, energy production, fiber mechanics, microcirculation, and electrical activation to the clinically observed global functions. These are measured by pressure, volume, shape, coronary flow, heart rate, and other changes. It is a complex interactive system requiring the intense efforts of capable scientists in the life sciences, including medicine, exact sciences, engineering and biomedical technology devoted to address these multivariable, multidisciplinary challenges, so as understand and control the pathologies involved. Here we present some of our past interactive studies and highlight two new models, one demonstrating micro to macro integration, and one involving tissue-organ interaction of various parameters. These models yield new insights into cardiac performance. PMID- 9330756 TI - Inhibition of virus-induced age-dependent poliomyelitis by interferon-gamma. AB - Age-dependent poliomyelitis (ADPM) is a neuroparolytic disease which results from combined infection of susceptible mice with lactate dehydrogenase-elevating virus (LDV) and murine leukemia virus (MuLV). The present study examined the effects of interferon-gamma (IFN-gamma) treatment on the incidence of ADPM, replication of LDV and MuLV and anti-LDV immunity. IFN-gamma treatment of ADPM-susceptible C58/M mice protected them from paralytic disease, but had no detectable effect on the IgG anti-LDV response or LDV viremia. IFN-gamma-mediated protection from ADPM correlated with reduced expression of LDV RNA, but not MuLV RNA, in the spinal cords of C58/M mice. These results confirm that spinal cord LDV replication is the determinant of ADPM and demonstrate that cytokine-mediated inhibition of LDV replication in the central nervous system prevents neuroparalytic disease. PMID- 9330758 TI - A rapid and quantitative assay for inhibition of 3' cleavage activity of HIV-1 integrase. AB - The human immunodeficiency virus-1 (HIV-1) integrase catalyzes the specific removal of two nucleotides at either 3' end of each long terminal repeat (LTR) sequence of the proviral DNA duplex. The most commonly used in vitro assays for integrase employ 5' end 32P-labeled double-stranded oligonucleotides and the product of integrase-associated endonuclease activity is visualized by denaturing gel electrophoresis followed by autoradiography. We report here a simple assay system based upon the liberation of [35S]GT dinucleotide from the 3' end of a double-stranded U5 LTR sequence derived from HIV-1. The uncleaved labeled substrate and the unlabeled large product are removed by adsorption to polyethyleneimine cellulose followed by centrifugation. The small labeled GT dinucleotide product released in the supernatant is quantitated in terms of counts released as a function of time. Since the method is rapid and quantitative, it should be useful in the kinetic evaluation of inhibitors of the 3' cleavage activity of HIV-1 integrase. PMID- 9330757 TI - Toxicity associated with high dosage 9-[(2R,5R-2,5-dihydro-5-phosphonomethoxy)-2 furanyl]adenine therapy off attempts to abort early FIV infection. AB - 9-[(2R,5R-2,5-dihydro-5-phosphonomethoxy)-2-furanyl]adenine, or D4API, was tested in the feline immunodeficiency virus (FIV) infection model and found to be significantly more inhibitory in vitro than its parent compound 9 phosphonylmethoxethyl adenine (PMEA). Cytotoxicity was less than for PMEA or azidothymidine (AZT) for culture periods of 7 days, but more toxic after 10 days. D4API was rapidly absorbed by cats following subcutaneous inoculation, with a plasma half-life of less than 1 h after intravenous inoculation and between 2 and 3 h after subcutaneous injection. Peripheral blood mononuclear cells collected from cats given a single dose of D4API were refractory, however, to FIV infection in vitro for up to 24 h. Given its prolonged intracellular phase and high selectivity index, high dose D4API therapy was tested for its ability to abort an acute (i.e. 2 week) FIV infection. A divided daily dose of D4API, which was one fourth the toxic dose and 125 times the concentration that would totally inhibit virus replication in vitro, completely abrogated the anticipated viremia and antibody responses. Unfortunately, a majority of treated/uninfected and treated/infected test cats died acutely of drug toxicity after 47 days of treatment. Toxicity in vivo mirrored what was observed in vitro, being precipitous and cumulative in nature. Toxic signs included widespread hepatic and lymphoid necrosis. A surviving treated/FIV infected cat remained healthy to day 175 when the study was terminated; antibodies appeared 2 months later than in untreated/infected cats and virus was only detectable at low levels on day 175. In contrast, untreated/infected cats were viremic and antibody positive from 3 to 4 weeks post-infection onwards. Therefore, it was possible to alter, but not abort, an early FIV infection with prolonged, high-dose D4API treatment. PMID- 9330759 TI - Phosphorylation of aciclovir, ganciclovir, penciclovir and S2242 by the cytomegalovirus UL97 protein: a quantitative analysis using recombinant vaccinia viruses. AB - We used recombinant vaccinia viruses (rVV) containing the UL97 open reading frame (ORF) of the human cytomegalovirus (HCMV) to investigate the UL97-dependent phosphorylation of different nucleoside analogs. The rVV T1 expressed the wild type UL97 protein whereas rVV A5 contained a 12 bp deletion in the UL97 which had been known to be responsible for resistance of HCMV to ganciclovir (GCV). The rVV T1opal was generated which contained a stop codon at position 1089 of the UL97 ORF and which expressed a truncated UL97 protein. We quantitatively analyzed the capability of these rVVs to phosphorylate GCV, penciclovir (PCV), aciclovir (ACV) and 2-amino-7-[(1,3-dihydroxy-2-propoxy)methyl] purine (S2242) as well as the natural nucleosides deoxycytidine and deoxythymidine. Moreover, we compared their phosphorylating capability with that of herpes simplex virus type 1 strains. In thymidine kinase (TK)-deficient 143B cells infected with rVV T1, the three compounds GCV, ACV and PCV were phosphorylated with different efficiency whereas in cells infected with the rVV A5 a markedly reduced but not completely abolished phosphorylation of these compounds was observed. In rVV T1opal-infected cells no specific phosphorylation of the compounds was detectable at all. Neither S2242 nor the natural substrates of TKs were phosphorylated by any of the vaccinia recombinants. The rVVs proved to be a suitable tool for analysis of UL97 dependent phosphorylation of nucleoside analogs and also allowed to quantitatively study the influence of UL97 mutations on drug phosphorylation. PMID- 9330761 TI - Antiviral activity of an extract from leaves of the tropical plant Acanthospermum hispidum. AB - Incubation of the alphaherpesviruses pseudorabiesvirus (PRV) and bovine herpesvirus 1 during infection of cell cultures with an extract prepared from the leaves of Acanthospermum hispidum impaired productive replication of these viruses in a concentration-dependent manner whereas propagation of classical swine fever virus, foot-and-mouth disease virus and vaccinia virus was not affected. The 50% inhibitory concentration for cell growth (IC50) was 107 +/- 5 microliters/ml, and the concentration reducing PRV yield by 1 log10 (90% effective concentration, EC90) was 8 +/- 3 microliters/ml. The selectivity index calculated as the IC50/EC90 ration was 13 +/- 4. Delineation of the mechanism of the antiviral activity demonstrated inhibition of alphaherpesvirus attachment to and, to a lesser extent, penetration into the cells. In contrast, viral gene expression was not inhibited by the extract when added after entry of virions into the target cells. Reduced antiviral activity of A.h. against PRV deletion mutants lacking glycoprotein C (gC) or glycoproteins gC, gE, gG and gI altogether indicated that gC alone and/or viral attachment complexes of which gC is a component constitute the target structures for A. hispidum. PMID- 9330760 TI - Alkoxy propane prodrugs of foscarnet: effect of alkyl chain length on in vitro antiviral activity in cells infected with HIV-1, HSV-1 and HCMV. AB - The identification of more effective and less toxic foscarnet (PFA) analogs for antiviral therapy would be useful. We recently synthesized 1-O-octadecyl-sn glycero-3-phosphonoformic acid (ODG-PFA) and noted a 93-fold increase in its anti HCMV activity relative to PFA. In addition, the antiviral activity of ODG-PFA in herpes simplex virus type-1 (HSV-1) and human immunodeficiency virus type-1 (HIV 1) infected cells was increased 40-fold relative to PFA (Hostetler et al., 1996. Antiviral Res. 31, 59). To evaluate structure-activity relationships further, we synthesized alkoxypropyl esters of foscarnet with varying alkyl chain lengths and degrees of saturation. These compounds were tested in vitro for activity and selectivity in comparison with PFA and ODG-PFA in cells infected with HCMV, HSV-1 or HIV-1. Antiviral activity was strongly dependent on chain length with alkyl ethers 14-18 carbon atoms long exhibiting the greatest antiviral activity against HCMV and HSV-1. In HIV-infected HT4-6C cells, optimal activity was observed at 18 22 carbon chain lengths. The antiviral activities of 1-octadecyloxypropane-3-PFA and 1-docosyloxypropane-3-PFA were 135- and 338-fold greater than that of PFA in HT4-6C cells infected with HIV-1. This also represents a 2.6-6-fold improvement in antiviral activity over ODG-PFA, the previously reported analog. PMID- 9330762 TI - The antiviral xanthate compound D609 inhibits herpes simplex virus type 1 replication and protein phosphorylation. AB - The mechanism of antiviral action of tricyclodecan-9-yl-xanthogenate (D609) was investigated in vitro. D609 inhibited herpes simplex virus type 1 (HSV-1) replication without apparent cytotoxicity. It reduced phosphorylation of virus infected cell polypeptides and inhibited the HSV-1 encoded protein kinase (US3 PK) and, to a lesser extent, cellular protein kinase C in vitro. Virus production was reduced by D609 at concentrations greater than 3.8 microM, with complete inhibition at 75.2 microM at an MOI of 1 PFU/cell or less. Addition of D609 could be delayed until 7 h post-infection and still inhibit virus replication. Phosphorylation of infected cell viral polypeptides of 34 (similar molecular weight to the substrate of the viral US3 protein kinase) and 69 kDa was inhibited at 18.4 microM. Treatment of infected or uninfected cells with 37.6 microM D609 reduced protein phosphorylation to background levels. A concentration of 1.9 microM D609 in vitro inhibited the viral US3-encoded PK, which had been purified from infected cell lysates by affinity chromatography and identified by specific antibody. Purified cellular protein kinase C was inhibited at 75.2 microM D609 whereas other cellular kinases including casein kinase 1 and cAMP dependent kinase were not inhibited at concentrations as high as 188 microM D609. Collectively these data indicate that the mechanism of antiviral action of D609 is by inhibition of protein kinases and protein phosphorylation affecting a late step in HSV replication. PMID- 9330777 TI - Pharmacokinetic analysis and antiepileptic activity of two new isomers of N valproyl glycinamide. AB - Valproyl glycinamide (TV 1901-VPGD) is a new antiepileptic drug, which is currently undergoing clinical trials. The present study explored the pharmacokinetics and pharmacodynamics (anticonvulsant activity and neurotoxicity) of two new isomers of valproyl glycinamide: valnoctyl glycinamide (VCGD) and diisopropylacetyl (DIGD). Both VCGD and DIGD showed anticonvulsant activity and a safety margin in mice similar to those of VPGD. Following i.v. administration (556 mg) to six dogs, VCGD had a clearance (Cl) value of 3.8 +/- 1.1 Lh-1 (mean +/- SD), a volume of distribution (Vss) of 15 +/- 2 L, and a half-life (t1/2) of 1.9 +/- 0.3 h. DIGD had Cl, Vss, and t1/2 values of 10 +/- 0.8 Lh-1, 19 +/- 3 L, and 1.6 +/- 0.2 h, respectively. Neither VCGD nor DIGD operated as chemical drug delivery systems (CDDSs) of glycine, valnoctic acid, or diisopropyl acetic acid and both showed antiepileptic profiles different from that of valproic acid (VPA). Both glycinamides were biotransformed to their glycine analogues with similar fractions metabolized (fm): 59 +/- 5% (VCGD) and 62 +/- 15% (DIGD). The two glycine metabolites, valnoctyl glycine (VCGA) and diisopropylacetyl glycine (DIGA), were also administered to the same dogs in order to calculate the above fm values. Both VCGA and DIGA had higher Cl and lower Vss values than VCGD and DIGD and therefore their mean t1/2 values were 0.43 +/- 0.02 and 0.30 +/- 0.07 h, respectively. VCGA and DIGA were excreted mainly intact in the urine, with fractions excreted unchanged (fe) of 60 +/- 9 and 55 +/- 7%, respectively. The improved pharmacokinetic profile of VCGD and DIGD relative to their glycine analogues may explain the similarity of their anticonvulsant activity to that of valproyl glycinamide. The current study demonstrates the benefit of the structure pharmacokinetic-pharmacodynamic relationship (SPPR) approach in developing and selecting a potent antiepileptic compound in intact animals based not only on its intrinsic pharmacodynamic activity but also on its improved pharmacokinetic profile. PMID- 9330778 TI - Integrated pharmacokinetic and metabolic modeling of selegiline and metabolites after transdermal administration. AB - Selegiline (SEL) is a selective, irreversible inhibitor of MAO-B, used in the treatment of Parkinson's disease, either alone or as an adjunct to L-DOPA. Selegiline hydrochloride (HCl) undergoes significant first-pass metabolism following oral administration. Transdermal delivery avoids the first-pass effect and provides greater and more prolonged levels of unchanged SEL and reduced levels of metabolites (N-desmethylselegiline (DES), L-amphetamine (AMP), and L methamphetamine (MET) compared to the oral regimen. An integrated pharmacokinetic metabolic model which predicts plasma concentrations of SEL and metabolites following a single 24 h application of a selegiline transdermal system (STS) is proposed. The model is based on the metabolic conversion of SEL to DES and MET and subsequently to AMP. The input function is described by a zero-order constant for the delivery of SEL from the STS system based on in vitro studies of penetration of SEL across human skin. The elimination-non-metabolic constants for each analyte account for the urinary elimination. Plasma concentration data from a pilot pharmacokinetic study in which six healthy male volunteers were administered single 24 h applications of a 1.8 mg cm2, 10 cm2 STS were used to examine this model. The coefficient of determination was 0.98 and model selection criterion was 3.4 for mean data fits, supporting the goodness of fit of the model. The pharmacokinetic parameters obtained by non-compartmental analysis were comparable to those predicted by a compartmental model. The model also predicted urinary recoveries for AMP and MET and negligible recovery for SEL and DES consistent with recent studies with the STS in which urine was collected. The metabolic conversion constant from SEL to DES was significantly lower than the conversion constant from SEL to MET, indicating that metabolism of SEL is primarily driven towards MET following transdermal administration. The metabolic conversion from MET to AMP was less than the conversion from DES to AMP. This simultaneous prediction of the SEL and metabolites is essential as the metabolic ratios have been linked to the neuroprotective effects of SEL. These findings support the proposed regional delivery advantage attributed to the transdermal route compared to the conventional therapy with the oral tablet. Future model applications may also help identify significant covariates (i.e. age, gender, and disease state) in upcoming clinical trials. PMID- 9330779 TI - Lack of effect of food on the steady state pharmacokinetics of BMS-181101, an antidepressant, in healthy subjects. AB - The effect of food on the pharmacokinetics of BMS-181101, a new anti-depressant under development, was investigated in 12 healthy male volunteers at steady state. Each subject received a 15 mg oral dose of BMS-181101 twice a day (q 12 h) for 11 days and a morning dose of BMS-181101 on day 12. Six subjects were randomly assigned to receive BMS-181101 under fasted conditions from days 1 to 6 and then crossed over to fed conditions from days 7 to 12. The other six subjects received the reverse conditions, fed for days 1-6 and fasted for days 7-12. Serial blood samples were collected up to 12 h on days 6 and 12 following the administration of the morning dose. In addition, trough blood samples were collected on days 4, 5, 10, and 11 prior to the morning dose. Plasma samples were analyzed for intact BMS-181101 using a validated high-performance liquid chromatography method with an electrochemical detector. BMS-181101 was well tolerated both with and without ingestion of food. The statistical evaluation of the Cmin values indicated that steady state of BMS-181101 was achieved by the fourth day of dosing regardless of whether the subject was fasted or fed. When BMS-181101 was administered with food, Cmax was reduced by about 25% and tmax was prolonged by 1 h. However, AUCtau, t1/2, and time to attain steady state of BMS 181101 were not altered by ingestion of food. In summary, BMS-181101 can be given with food without adversely impacting the safety or pharmacokinetic profiles of the drug. PMID- 9330780 TI - Effects of the rate and composition of fluid replacement on the pharmacokinetics and pharmacodynamics of intravenous azosemide. AB - The effects of differences in the rate and composition of intravenous fluid replacement for urine loss on the pharmacokinetics and pharmacodynamics of azosemide were evaluated using rabbit as the animal model. Each rabbit received a 4h constant intravenous infusion of 1 mg kg-1 azosemide with 0% replacement (treatment I, n = 4), 50% replacement (treatment II, n = 5), and 100% replacement (treatment III, n = 5) with lactated Ringer's solution, as well as with 100% replacement with 5% dextrose in water (D-5-W, treatment IV; n = 5). Renal clearance and urinary excretion rate of the drug in treatment III were considerably higher than those in treatments I, II, and IV. In spite of the similarities in kinetic properties, diuretic and/or natriuretic effects of azosemide were markedly different among the four treatments. For example, the mean 8 h urine output values were 98.2, 178, 733, and 237 mL for treatments I-IV, respectively, and the corresponding values for sodium excretion were 11.1, 19.4, 76.4, and 14.2 mmol, and for chloride 13.4, 23.8, 78.9, and 17.1 mmol. Except for treatment III, diuresis and/or natriuresis were found to be time dependent, generally decreasing with time until reaching a low plateau during the later hours of infusion. The present findings also show that (i) no fluid replacement and 100% replacement with D-5-W both produce the same degree (not significantly different) of severe acute tolerance in natriuresis, indicating the insignificance of water compensation in tolerance development; (ii) in treatment II, where neutral sodium balance was achieved, the development of acute tolerance in diuresis can mainly be attributed to negative water balance under this special condition; and (iii) at steady state the hourly diuresis and natriuresis can differ up to about 6.87- and 5.21-fold between treatments. Some implications for the bioequivalence evaluation of dosage forms of azosemide are discussed. PMID- 9330781 TI - Transport properties of 3'-azido-3'-deoxythymidine and 2',3'-dideoxyinosine in the rat choroid plexus. AB - Transport properties of 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (DDI) were characterized in the isolated rat choroid plexus. AZT and DDI competitively inhibited the active transport of [3H]benzylpenicillin, a prototypic organic anion, with Ki values of 85.4 +/- 13.1 and 155 +/- 22 microM, respectively. Accumulation of [3H]DDI was against an electrochemical potential via a saturable process (K(m) = 29.7 +/- 4.9 microM, Vmax = 13.5 +/- 2.4 pmol min 1/microL tissue) that was inhibited by metabolic inhibitors (carbonylcyanide p trifluoromethoxyphenylhydrazone, 10 microM, and rotenone, 30 microM) and sulphydryl reagents (p-chloromercuribenzoic acid, 100 microM, and p chloromercuribenzenesulphonic acid, 100 microM), but did not require an inwardly directed Na+ gradient. Accumulation of [3H]DDI was inhibited by benzylpenicillin and AZT in a dose-dependent manner, with IC50 values of 91.6 +/- 28.9 and 294 +/- 84 microM, respectively. In contrast, no significant accumulation of [3H]AZT was observed. These results suggest that DDI is transported, at least in part, by the transport system for organic anions located on the rat choroid plexus, whereas AZT is recognized, but not transported by this system. PMID- 9330782 TI - Pharmacokinetic and local tissue disposition studies of naproxen-following topical and systemic administration in dogs and rats. AB - The pharmacokinetic profiles of naproxen in blood and synovial fluid (SF) following topical and i.v. bolus administration in dogs, and the local tissue disposition of the drug following topical and oral administration in rats, were investigated to assess the feasibility of topical delivery of naproxen for local and systemic effects. The naproxen gel in poloxamer 407 (PF-127) was applied on the stifle joint of dogs, and serum and synovial fluid samples were collected. For local tissue disposition studies, the naproxen gel was applied on the dorsal skin in rats, and blood, skin, and muscle samples were taken at 3, 6, and 12 h postdose after removing the residual gel from the skin. Steady state serum concentrations occurred at approximately 20 h after topical doses and lasted for the next approximately 30 h in dogs. Similar SF-serum concentration ratios of naproxen were found between i.v. (0.61 +/- 0.16) and topical (0.55 +/- 0.14) routes of administration. Following the i.v. dose, the half-life of naproxen in SF (approximately 60 h) was significantly longer than that in serum (approximately 40 h). The bioavailability of naproxen in the topical gel was approximately 2% of the applied dose in dogs. A large accumulation of drug in the epidermis, dermis, and muscle tissue beneath the gel application site was found in rats. Isopropyl myristate (IPM) significantly increased the systemic absorption as well as the concentrations of naproxen in the underlying dermis and muscle tissues, but exerted little effect on the disposition of naproxen in the epidermis. PMID- 9330784 TI - Effect of food on the bioavailability of fexofenadine hydrochloride (MDL 16455A). PMID- 9330783 TI - Pharmacokinetics and dose proportionality of beclomethasone from three strengths of a CFC-free beclomethasone dipropionate metered-dose inhaler. AB - As part of a development program to offer alternatives to chlorofluorocarbon (CFC) containing metered-dose inhalers, beclomethasone dipropionate has been formulated in a CFC-free system at three strengths: 50, 100, and 200 micrograms/actuation ex valve. To measure serum levels and dose proportionality of the beclomethasone derived from beclomethasone dipropionate, 13 mild to moderate asthmatic patients received a single dose of eight inhalations from each strength according to a double-blind crossover design. Seven patients were studied over 4 h and six patients over 12 h. For the total doses of 400, 800, and 1600 micrograms studied over 12 h, Cmax and AUC increased in a ratio of 1:1.8:3.1. A good correlation was seen between the fine-particle mass delivered and the in vivo performance of the three strengths. From a clinical point of view, the predictable increases in serum levels with an increase in dose will permit the clinician to effectively titrate a patient with this product. PMID- 9330785 TI - Synergism between hypotonically induced calcium release and fatty acyl-CoA esters induced calcium release from intracellular stores. AB - The non-mitochondrial Ca2+ stores in permeabilized A7r5 cells responded to a decrease in Mg-ATP concentration with a pronounced Ca2+ release if 20 microM CoA was present. This release was rather specific for the preincubation or removal of ATP. ATP gamma S was much less effective and AMP-PNP, GTP, ITP, CTP, UTP, ADP, AMP, adenosine and adenine had no effect. CoA activated with an EC50 of 6 microM. Dephospho-CoA was a less effective cofactor and desulfo-CoA was ineffective. The release induced by Mg-ATP removal did not occur in the presence of 2% fatty acid free bovine serum albumin and did not develop at 4 degrees C. All these findings suggest that CoA had to be acylated by endogenous fatty-acyl-CoA synthetase to become effective. Myristoyl- and palmitoyl-CoA esters were identified as the most effective cofactors for the release. Ca2+ release induced by removing Mg-ATP did not occur if the osmolality of the medium was kept constant by addition of mannitol, sucrose, KCl, MgCl2 or Mg-GTP, indicating that the decrease in tonicity was the trigger for the release. Mg-ATP plus CoA also synergized with Ca2+ release induced by a hypotonic shock imposed by diluting the medium with H2O. Osmolality changes induced by decreasing the Mg-ATP concentration were more effective in releasing Ca2+ than equal decreases in concentration of all solutes. We conclude that fatty acyl-CoA esters sensitize the hypotonically induced Ca2+ release from the non-mitochondrial Ca2+ stores. PMID- 9330786 TI - Co-ordinated control of apical calcium influx and basolateral calcium efflux in rabbit cortical collecting system. AB - Transcellular Ca2+ transport in the distal nephron involves passive Ca2+ influx at the apical membrane, diffusion through the cytosol and active extrusion across the opposing basolateral membrane. The molecular identity of the apical Ca2+ entry step is still elusive, but its regulatory aspects have been analyzed in the present study. To this end, rabbit connecting and cortical collecting tubular cells were cultured on permeable and transparent supports and the apical Ca2+ influx was deduced from Mn2+ quenching of Ca2+ independent Fura-2 fluorescence, while the intracellular Ca2+ concentration ([Ca2+]i) was measured simultaneously. In parallel experiments, transcellular Ca2+ transport was determined isotopically as 45Ca2+ flux from the apical to basolateral compartment. Decreasing the apical pH from 7.4 to 5.9 inhibited transcellular Ca2+ transport by 53 +/- 1%, whereas apical Ca2+ influx was reduced by 39 +/- 7% and [Ca2+]i decreased by 18 +/- 3%. Reversal of the Na+/Ca2+ exchanger by iso-osmotic replacement of Na+ by N-methyl D-glucamine in the basolateral compartment resulted in 50 +/- 5% inhibition of Ca2+ transport, 46 +/- 3% reduction of apical Ca2+ influx and 60 +/- 3% increase in [Ca2+]i. In the absence of basolateral Ca2+, however, this manoeuvre decreased [Ca2+]i by 21 +/- 8%, while Ca2+ transport and apical Ca2+ influx were reduced by the same magnitude as in the presence of Ca2+, that is by 53 +/- 6% and 45 +/- 4%, respectively. Stimulation of adenylyl cyclase with forskolin (10(-5) M) increased transcellular Ca2+ transport by 108 +/- 40%, stimulated apical Ca2+ influx by 120 +/- 17% and increased [Ca2+]i by 110 +/- 2%. In conclusion, the apical Ca2+ influx is regulated by apical pH, intracellular cAMP and basolateral Na+/Ca2+ exchanger activity, and is coupled in an 1:1 fashion to the rate of transepithelial Ca2+ transport. PMID- 9330787 TI - Kinetics of calcium steps underlying calcium oscillations in melanotrope cells of Xenopus laevis. AB - Melanotrope cells of Xenopus laevis display intracellular calcium oscillations which are generated at the plasma membrane and travel as a wave through the cytoplasm into the nucleus. An oscillation involves discrete increases in intracellular Ca2+ ('steps'), followed by a relatively smooth return to the basal Ca2+ level. The aim of our investigation was to determine what role these steps play in shaping the Ca2+ signal in melanotrope cells, by conducting a high resolution spatio-temporal analysis of the kinetics of the Ca2+ steps. To this end Fura-red loaded cells were analysed by confocal laser scanning microscopy using the line scanning method to achieve 6 ms time resolution. Furthermore, the kinetics of the steps were analysed in 3 different intracellular areas, to see if there are spatial differences in Ca2+ signalling kinetics. The results showed that each calcium oscillation is built up by 3-4 steps that were generated very quickly and had approximately the same size. Following each Ca2+ step, there was a slow removal of calcium before the next step boosted the overall level of Ca2+. Since the Ca2+ steps were most pronounced directly beneath the plasma membrane, they appear to be generated in this region. The speed of the Ca2+ wave near the membrane exceeded 40 microns/s, indicating an active mechanism for wave propagation. In deeper regions of the cell, the wave speed was much slower (about 8 microns/s) and the size of each step was smaller, indicating that regulation occurs within a narrower range of [Ca2+]i. Inside the nucleus, however, the calcium wave accelerated again (23 microns/s). Treatment with TRH evoked a high amplitude Ca2+ transient and increased the number of Ca2+ steps to 5 or 6. Each step had approximately the same size as the steps of the pretreatment Ca2+ oscillations. Caffeine treatment, which increased the frequency of the oscillations, had no effect on the number or the size of the Ca2+ steps, but it reduced the time needed for each step to reach its maximum height. We suggest a possible 'building block' function for the Ca2+ steps, whereby a cell generates more steps to achieve a high oscillation amplitude or accelerates the speed of the steps to increase the frequency of oscillations. Both phenomena may play a crucial role in the encoding of information transduced from an extracellular input to the intracellular target. PMID- 9330788 TI - Arachidonic acid mediates calcium influx induced by basic fibroblast growth factor in Balb-c 3T3 fibroblasts. AB - Basic fibroblast growth factor (bFGF), a peptide acting as a mitogen in different cell types, is able to induce a long lasting non capacitative calcium influx from the extracellular medium in Balb-c 3T3 mouse fibroblasts. This effect is mediated by the tyrosine kinase activity of bFGF receptors and the opening of voltage independent, agonist activated calcium channels. In this paper we investigate the signal transduction steps involved in this process using single cell calcium fluorimetry and electrophysiological techniques. One of the pathways initiated by the binding of growth factors to their tyrosine kinase receptors is the activation of cytosolic phospholipase A2 (cPLA2) and the release of arachidonic acid (AA) from the plasma membrane with the subsequent production of eicosanoids. We show here that, in our preparation, this pathway is involved in the opening of the bFGF-activated calcium permeable channels, through the activation of mitogen activated protein kinase (MAPK) and cPLA2. Evidence for direct involvement of AA is given by the finding that: (i) bFGF induces AA release from Balb-c 3T3 cells; (ii) blockers of AA metabolism are not effective; and (iii) the application of either arachidonic acid or its non metabolizable analogue 5,8,11,14 eicosatetraynoic acid (ETYA) reproduces the responses described for bFGF. Finally, single channel analysis indicates that bFGF, AA and ETYA can activate the same calcium permeable channel. PMID- 9330789 TI - Shear-stress causes polarized change in cytoplasmic calcium concentration in human umbilical vein endothelial cells (HUVECs). AB - Using a newly developed, parallel-plate flow-chamber for confocal laser scanning microscopy (CLSM), we studied the distribution and temporal changes in intracellular Ca2+ concentration ([Ca2+]i) in individual HUVECs stimulated by shear-stress. In the presence of ATP, shear-stress (1-10 dyne/cm2) caused a rise in [Ca2+]i, whereas no such response was observed in the absence of ATP or in the presence of Ni2+, a nonspecific, plasma membrane Ca2+ channel blocker. These results suggest that both ATP and Ca2+ influx are essential for the increase in [Ca2+]i in response to shear stress at less than 10 dyne/cm2. Analysis of [Ca2+]i distribution revealed a repetitive intracellular 'Ca2+ wave' originating from the upstream edge of the cell in some populations of HUVECS, which was transmitted to the downstream of the cell. The polarized [Ca2+]i response induced by shear stress might be integral to polarized cellular reactions such as remodeling of endothelial lining. PMID- 9330790 TI - Synchronized calcium spiking resulting from spontaneous calcium action potentials in monolayers of NRK fibroblasts. AB - The correlation between the intracellular Ca2+ concentration ([Ca2+]i) and membrane potential in monolayers of density-arrested normal rat kidney (NRK) fibroblasts was investigated. Using the fluorescent probe Fura-2, spontaneous repetitive spike-like increases in [Ca2+]i (Ca2+ spikes) were observed that were synchronised throughout the entire monolayer. Ca2+ spikes disappeared in Ca(2+) free solutions and could be blocked by the L-type Ca2+ channel antagonist felodipine. Simultaneous measurements of [Ca2+]i and membrane potential showed that these Ca2+ spikes were paralleled by depolarisations of the plasma membrane. Using patch clamp measurements, action potential-like depolarisations consisting of a fast spike depolarisation followed by a plateau phase were seen with similar kinetics as the Ca2+ spikes. The action potentials could be blocked by L-type Ca2+ channel blockers and were dependent on extracellular Ca2+. The plateau phase was predominantly determined by a Cl- conductance and was dependent on intracellular Ca2+. The presence of voltage-dependent L-type Ca2+ channels in NRK cells was confirmed by patch clamp measurements in single cells. It is concluded that monolayers of density-arrested NRK fibroblasts exhibit spontaneous Ca2+ action potentials leading to synchronised Ca2+ spiking. This excitability of monolayers of fibroblasts may represent a novel Ca2+ signaling pathway in electrically coupled fibroblasts, cells that were hitherto considered to be inexcitable. PMID- 9330791 TI - Novel fluorescent indicator proteins for monitoring free intracellular Ca2+. AB - We have recently described a fluorescent indicator protein in which red- and blue shifted variants of green fluorescent protein are joined by the calmodulin binding sequence from smooth muscle myosin light chain kinase [Romoser V.A., Hinkle P.M., Persechini A. Detection in living cells of Ca(2+)-dependent changes in the fluorescence of an indicator composed of two green fluorescent protein variants linked by a calmodulin-binding sequence. A new class of fluorescent indicators. J Biol Chem 1997; 272: 13270-13274]. The fluorescence emission of this protein at 505 nm (380 nm excitation) is reduced by approximately 65% when (Ca2+)4-calmodulin is bound, with a proportional increase in fluorescence emission at 440 nm. We have found that fusion of an engineered calmodulin, in which the C- and N-terminal EF hand pairs have been exchanged, to the C-terminus of this protein results in a novel indicator that responds directly to changes in the Ca2+ ion concentration, with an apparent Kd value of 100 nM for Ca2+ in the presence of 0.5 mM Mg2+. The affinity of the indicator for Ca2+ can be decreased by altering the amino acid sequence of the calmodulin-binding sequence to weaken its interaction with the intrinsic calmodulin domain. The fluorescence emission of this indicator can be used to monitor physiological changes in the free Ca2+ ion concentration in living cells. PMID- 9330793 TI - Adult outcome of children reared for long-term periods in foster families. AB - OBJECTIVE: To study the long term impact of adverse childhood experiences resulting from family breakdowns combined with a stable care environment. Another aim was to determine predictive factors for maladjusted psycho-social integration in adulthood. METHOD: Sixty-three children from severely psychosocially dysfunctioning families selected from among those having been in care in an institutional setting: All had been reared for at least 5 years by foster families, had been out of care for more than 5 years and were at least 23 years old at the time of the survey. Semi-structured interviews were used in a follow up study to assess adult outcome, essentially in terms of professional status, social, and family relationships. RESULTS: Data was obtained for 94% of the study population (n = 59), 71% via direct interviews (n = 45). The majority had managed to overcome their childhood adversities: 56% were well-integrated socially, 12% had average integration results, 20% were partially integrated and 10% were in situations of failure. These difficulties were linked to multiple family disturbances and repeated traumatic experiences during childhood (p < .05). Multiple regression analyses indicated that these risk factors accounted for 28% of the variance in the social integration score (p < .0001). Severe emotional deprivation over a prolonged period was a contributing factor to clinical disorders. CONCLUSION: At the study period, intergenerational repetition of "child placement" behaviors, significant in the previous generation, had practically disappeared. The results also highlighted the substantial psychotherapeutic and child-rearing assistance provided by the staff of the foster care agency. PMID- 9330792 TI - nNOS and Ca2+ influx in rat pancreatic acinar and submandibular salivary gland cells. AB - Regulation of agonist-activated Ca2+ influx by the NOS pathway through generation of cGMP is being found in an increasing number of cell types. In the present work, we examined the role of the NOS pathway in agonist-evoked [Ca2+]i oscillations and attempted to identify the NOS isoform most likely to regulate Ca2+ influx. For this, we first show that two Ca(2+)-mobilizing agonists acting on pancreatic acinar cells, bombesin (BS) and the cholecystokinin analog CCK-JMV 180 (CCKJ), evokes different type of [Ca2+]i oscillations. The BS-evoked [Ca2+]i oscillations rapidly became acutely dependent on the presence of extracellular Ca2+, whereas the CCKJ-evoked oscillations continue for long periods of time in the absence of Ca2+ influx. This differential behavior allowed us to isolate Ca2+ influx and study its regulation while controlling for non specific effects on all other Ca2+ transporting events involved in generating [Ca2+]i oscillations. Inhibitors of selective steps in the NOS pathway inhibited agonist-induced cGMP production. The inhibitors were then used to show that scavenging NO with reduced hemoglobin, inhibition of guanylyl cyclase with 1H-[1,2,4] oxadiazolo[4,3-a] quinoxaline-1-one (ODQ) and inhibition of protein kinase G with Rp-8-pCPT-cGMPS inhibited [Ca2+]i oscillations evoked by BS but not those evoked by CCKJ. These findings were extended to duct and acinar cells of the SMG. In these cells, Ca(2+)-mobilizing agonists stimulate large Ca2+ influx, which was inhibited by all inhibitors of the NOS pathway. Western blot analysis and immunolocalization revealed that the cells did not express iNOS, eNOS was expressed only in blood vessels and capillaries whereas nNOS was expressed at high levels next to the plasma membrane of all cells. Accordingly, the nNOS inhibitor 7-nitroindazole (7 NI) inhibited BS- but not CCKJ-evoked [Ca2+]i oscillations and Ca2+ influx into SMG acinar and duct cells. Thus, together, our findings favor nNOS as the isoform activated by the Ca2+ released from internal stores to generate cGMP and regulate Ca2+ influx. PMID- 9330794 TI - Restricting the time of injury in fatal inflicted head injuries. AB - OBJECTIVE: To determine the normal clinical progression of fatal head injuries in children. Such information can then be used to estimate the time of injury in cases with obscure histories and will thus aid investigations of nonaccidental trauma. METHOD: A retrospective chart review design was used. One hundred and thirty eight accidental fatalities involving head injury were identified and 95 of these were used as the study group. Details of the cases were reviewed and cases in which a child either had a Glasgow Coma Scale (GCS) of 14-15 or was described as having a "lucid interval" or as being "conscious" were further studied. RESULTS: One "lucid interval" case was identified. This case involved an epidural hematoma. Three other cases that partially met the criteria for a lucid interval were also identified; one of these cases did not meet the criteria for inclusion in the study group. Review of head CTs revealed that brain swelling could be detected as early as 1 hour and 17 minutes post injury. CONCLUSIONS: The children studied were in obvious serious medical condition from the time of injury until death. If a history purports a lucid interval in a fatal head injury case that does not involve an epidural hematoma, that history is likely false and the injury is likely inflicted. The time of most fatal head injury events can be restricted to the time period after the last confirmed period of wellness for the child. In addition, the presence of brain swelling on a head CT scan is not helpful in restricting the time of injury. PMID- 9330795 TI - Anxiety, depression, and dissociation in women health care providers reporting a history of childhood psychological abuse. AB - OBJECTIVE: The purpose of this study was to explore the relationship between childhood psychological maltreatment and adult manifestations of depression, anxiety, and dissociation. METHOD: Women health care professionals reporting a history of childhood psychological maltreatment (n = 55) were compared to a nonabused control group (n = 55) on the three dimensions of anxiety, depression, and dissociation. The Childhood Experiences Questionnaire, a measure constructed specifically for this study to assess abuse history, was used to determine group membership. Participants were administered the State-Trait Anxiety Inventory (STAI), the Beck Depression Inventory (BDI), and the Dissociative Experiences Scale (DES). RESULTS: A significant discriminant function analysis using the STAI, BDI, and DES as predictor variables was able to correctly classify 74.5% of the psychologically abused participants and 89.1% of the nonabused group, with an overall hit rate of 81.8%. Statistically significant differences were obtained between the abused and nonabused groups on the STAI, BDI, and DES. CONCLUSIONS: Interpretation of these results suggests that participants who reported a history of childhood psychological abuse suffer significantly higher levels of depression and anxiety, and more frequent dissociative experiences, than the nonabused women. PMID- 9330796 TI - Child sexual revictimization by multiple perpetrators. AB - OBJECTIVE: The objectives of this study were to describe feelings, disclosure characteristics, family dysfunction, and health risky behaviors in those adolescents having unwanted sexual experiences (USE; any kind of sexual touching that was bad, uncomfortable, or forced) with multiple perpetrators and to compare these parameters with those adolescents having USE(s) with single perpetrators. METHOD: A cross-sectional survey of consecutive waiting room patients from four clinic sites was done in 538 adolescents and young adults; 76% of the study population were Hispanic and over half were poor. One hundred sixty-one subjects with single perpetrator USE(s) were compared with 97 subjects who had USE(s) with more than one perpetrator. RESULTS: Victims of multiple perpetrators were more likely than victims of single perpetrators to react with self-blame and delay disclosure of USE due to shame. When compared with victims of single perpetrators, those with multiple perpetrators were more likely to disclose their USE to protect self or others or because they became weary or intolerant of the abuse. Although family violence and substance abuse were common in both victims of single and multiple perpetrators of USE, these factors appeared to potentiate the likelihood of repeated victimization in childhood. Prevalence of health risky behaviors did not differ between the two groups. CONCLUSIONS: The findings indicated that sexual revictimization by multiple perpetrators is not uncommon and suggest that abused children should be questioned about this possibility. Children and teenagers who have USE(s) with more than one perpetrator may have more difficulties with psychological recovery due to increased shame and self blame. PMID- 9330797 TI - Pattern of child sexual abuse by young aggressors. AB - OBJECTIVE: The aim of the study was to determine whether sexual victimization of children by young aggressors differs from adult aggressors. METHOD: A case review was performed on medical records of children less than 12 years of age referred in 1992 to the Child Protection Clinic at a tertiary care pediatric hospital. RESULTS: Medical evaluation for sexual abuse was carried out on 316 children, 79% girls, 21% boys, mean age 6 +/- 2.7 years. Among known perpetrators, 39 were less than 16 years and 15 were between 16 and 19 years old. Young aggressors were more likely to abuse older female victims (p = .0009). They also were reported to engage in more genital/genital and genital/anal acts (p < .001). The aggressor's young age was found to be an important determinant related to a history of penetrative forms of sexual abuse (OR = 4.015, 95% C.I. 2.0581; 7.8319). Genital examination was specific for abuse (Adam's Class IV or V) in only 6.3% of victims, but significantly more often when the perpetrator was between 16-19 years old (p = .003). CONCLUSIONS: Adolescent aggressors appear to engage in more genital/genital and genital/anal sexual abuse than older aggressors. Victims of aggressors age 16 to 19 had a higher risk of having specific findings on the anal/genital examination. PMID- 9330798 TI - Prevention of child sexual abuse victimization: a meta-analysis of school programs. AB - OBJECTIVE: The aim of this article was to provide data about the effects of child sexual abuse prevention programs. A more specific aim was to estimate the contribution of potential moderator variables such as age, program duration, or sample size to effect size. METHOD: A meta-analytic approach was used to calculate post-test and follow-up effect sizes of 16 evaluation studies of school programs aimed at the prevention of child sexual abuse victimization. Tests of categorical models were used in the analysis of moderator variables. Multiple regression analysis was used to determine their association with effect sizes. RESULTS: Significant and considerable mean post-intervention (d = .71) and follow up (d = .62) effect sizes were found, indicating that victimization prevention programs are successful in teaching children sexual abuse concepts and self protection skills. Intervention characteristics such as duration and content of the program, and child characteristics such as age and SES were important moderators of effect size. CONCLUSIONS: Our findings corroborate and refine the positive conclusions of traditional narrative reviews. Programs that focus on skill training, allowing sufficient time for children to integrate self protection skills into their cognitive repertoire, are to be preferred. Future evaluation research should focus on transfer of training. PMID- 9330799 TI - Assessment issues and long-term effects of childhood abuse and neglect. PMID- 9330800 TI - The polygraph, its use in cases of alleged sexual abuse: an exploratory study. AB - OBJECTIVE: The study's objective was to examine the relationship of polygraph findings to other indices of likelihood of sexual abuse and to case decisions by prosecutors, child protection workers, and professional evaluators. METHOD: This is an exploratory study of 42 cases with sexual abuse allegations and polygraph results. Case record data were abstracted and coded, including polygraph results, child interviews, medical examinations, protective services records, police investigations, and professional evaluations. Descriptive statistics, bivariate analyses, and multivariate analyses were employed. RESULTS: Polygraph findings were unrelated to other evidence of likelihood of sexual abuse, that is to the child's statements or demonstrations of sexual abuse, medical evidence, psychological symptoms, or indicators of sexual abuse from sources other than the child. When alleged offenders passed polygraphs, criminal prosecution was not sought. However, failing polygraphs was not predictive of criminal prosecution. Decisions by child protective services to substantiate or not were weakly related to polygraph findings and consistently related to any indicators of possible sexual abuse. Decisions by professional evaluators about sexual abuse were best predicted by children's psychological symptoms. CONCLUSIONS: The findings reinforce already expressed reservations about the polygraph's utility in sexual abuse decision-making. Additional research is needed on decisions about the likelihood of sexual abuse. PMID- 9330802 TI - Childhood attachment and abuse: long-term effects on adult attachment, depression, and conflict resolution. AB - OBJECTIVE: The primary aim was to determine the relative contributions of early attachment and abuse history to adult attachment, depression, and conflict resolution behaviors. Differences between abused and nonabused respondents were also assessed. METHOD: A multi-scale questionnaire was completed by 879 college students. Hierarchical regression analyses were used to answer the primary research question, and analyses also compared the 26.4% of respondents who reported childhood abuse with those who did not. RESULTS: Respondents who indicated they had been abused as children reported less secure childhood and adult relationships than their nonabused counterparts. They were also more depressed and more likely to use destructive behaviors in conflict situations. Although both adult romantic attachment and respondents' depression scores were best accounted for by childhood attachment to mother and father rather than abuse history, the opposite pattern of results emerged for conflict resolution behaviors. In this case, abuse history was the stronger predictor, and parental attachment did not account for any significant additional variance. CONCLUSIONS: Results suggest that the long-term impact of childhood abuse may be mediated by early attachment experiences, whereas the long-term impact of abuse on conflict resolution behaviors may be considerably more direct. PMID- 9330801 TI - The Whiplash Shaken Infant Syndrome: has Caffey's syndrome changed or have we changed his syndrome? AB - OBJECTIVE: The aim of this study is to examine the data used by John Caffey in his description of the Whiplash Shaken Infant Syndrome and compare it with recent data in an attempt to determine whether the syndrome that he described has changed, or if we have changed his syndrome into what we now call The Shaken Infant Syndrome. METHOD: This study examined recent literature describing the Shaken Infant Syndrome, and compared it to Caffey's descriptions. In addition, a retrospective review of 71 children under the age of 3 years identified as having a subdural hematoma caused by other than accidental means during 54 months was done. This data was compared to data from the 27 case examples offered by Caffey in 1972 and his other descriptions in 1974 and 1946. RESULTS: A review of recent literature shows that our definition of Shaken Infant Syndrome today includes cases where impact trauma was involved. In contrast to Caffey's descriptions, we found the perpetrator to be more often male, fractures to be more often to ribs rather than long bones, and admissions of shaking and other trauma more often made. CONCLUSIONS: Our findings demonstrate that not only have we changed the diagnostic parameters from Caffey's original Whiplash Shaken Infant Syndrome, but the syndrome has also changed to reflect changes in medical diagnosis and in our society. PMID- 9330803 TI - The preparticipation sports examination for high school and college athletes. AB - The PSE can be used as a tool to allow athletes to participate safely in sports. The goal of the PSE is not to disqualify athletes but to ensure that their participation in sports does not unnecessarily increase their risk of injury. The PSE is most effectively conducted by the station method with multiple examiners, one of whom should have specialty training in musculoskeletal disorders. The examination should be conducted 6 weeks prior to the beginning of the season and at the beginning of each new level of competition, unless directed differently by local laws. The correct use of the PSE should screen for signs and symptoms of pathological states that may lead to a nontraumatic death while participating in sports. An effective musculoskeletal examination should detect any postinjury deficits that may lead to subsequent reinjury later in the season. It is our hope that a PSE, based on the literature, can be used to prevent some of the nontraumatic deaths and musculoskeletal injury associated with sports participation. PMID- 9330805 TI - Pediatric and adolescent sports injuries. AB - Scholastic-age sports are generally safe, and major musculoskeletal injuries are uncommon. Injuries are proportional to the athlete's age, size, and type of sport. Significant knee injuries do occur, especially during adolescence, and they require prompt, accurate diagnosis and specific treatment. The team physician plays a major role in injury prevention and management. PMID- 9330804 TI - The female athlete. AB - Women's participation in sporting activities is now diverse with new opportunities arising yearly. As a result, care of the the female athlete's unique medical concerns has become an important challenge and issue to the primary care physician. The major focus when caring for the female athlete should be the diagnosis and treatment of the female athlete triad. The components of the triad--disordered eating, amenorrhea, and osteoporosis--can have serious implications for the health of the female athlete. Appropriate prevention and screening methods for early diagnosis of the female athlete triad require future study and improvement. Healthy pregnant, postpartum, and breastfeeding women can continue to maintain physical activity. Musculoskeletal injuries from sports are, in general, not gender specific but are more often sport specific. One exception is the increased prevalence of anterior cruciate ligament injuries occurring in women soccer and basketball players. The exact cause of this is unknown but is continuing to be investigated. PMID- 9330806 TI - Common medical problems in sports. AB - This article reviews the most common medical problems encountered in the day-to day care of athletes at all levels of competition. Common medical conditions affecting the pulmonary, gastrointestinal, urological, and endocrine systems are reviewed, as well as common infectious diseases. Review of environmental factors affecting athletes, including sleep disorders, travel, and exposure to the environment during athletic competition, are discussed. PMID- 9330807 TI - Shoulder injuries in the athlete. AB - Musculoskeletal injuries constantly provide challenges to the team physician, including those to the shoulder. Shoulder injuries are common in athletes, whether as a result of direct contact from a collision or from repetitive overhead motion. This article reviews sports-related injuries to the shoulder, including similarities between sports, clinical evaluation, and rehabilitation of the athletes. PMID- 9330808 TI - The elbow. AB - Elbow disorders in the athletic population comprise a wide range of injuries from acute trauma to those caused by chronic overuse of the joint. Certain injuries are orthopedic emergencies that must be recognized immediately by the team physician to avoid potential complications. Other overuse injuries need to be accurately diagnosed and treated so further injury can be prevented and the athlete can return to competition as expediently as possible. Finally, the decision to refer an athlete for surgical treatment often rests with the team physician; only with an adequate understanding of the elbow disorders in the athlete can these decisions be made. PMID- 9330809 TI - Primary care of hand and wrist athletic injuries. AB - Athletic injures to the hand and wrist can range from simple sprains to severe fractures or soft-tissue disruptions that can permanently threaten the normal function of the extremity. This article deals with some of the more commonly noted sports-related injures to the hand and wrist to help the team physician make the correct diagnosis and establish the most effective treatment plan, so that the athlete may achieve maximum results and ultimately return to full participation in their sport. PMID- 9330810 TI - The athlete's heart. AB - We have provided an overview of the athlete's heart, focusing on the young athlete. Primary caretakers of athletes should know the major causes of exercise related cardiac complications and sudden cardiac death and look for these conditions during preparticipation evaluations. We strongly suggest that coaches and other athletic personnel be required to learn basic life support measures such as cardiopulmonary resuscitation (CPR) and to update their skills on an annual basis. Such efforts will help prevent additional exercise-related cardiac deaths. PMID- 9330811 TI - Thoracoabdominal injuries in the athlete. AB - Although thoracoabdominal injuries are uncommon in the athlete, they can be catastrophic if unrecognized or if diagnosis and treatment are delayed. This article reviews thoracic, intrathoracic, abdominal, and groin injuries in the athlete, and how they can be diagnosed and managed. PMID- 9330813 TI - Occupational allergic contact dermatitis from carnosol, a naturally-occurring compound present in rosemary. AB - A 56-year-old man, working in a food processing factory, developed contact dermatitis of his hands, forearms, and face after the introduction of a new herb extract (Rosmanox) made from the leaves of rosemary (Rosmarinus officinalis). He reacted to carnosol, the main constituent of Rosmanox. 226 controls were negative. To our knowledge, this is the 1st reported case of contact dermatitis from carnosol. PMID- 9330812 TI - Psychosocial factors in sports injury rehabilitation. AB - The psychology of sports injury rehabilitation is a relatively new field, even in comparison with the relatively youthful disciplines from which it has evolved. Although the psychology of sports injury has made a significant impact on the sports medicine team, the practical aspects of how and when to refer patients to psychologists need to be better understood. A recent survey of 20 sports medicine physicians indicated a high degree of psychological or behavioral concerns occurring in conjunction with sport injuries, and an increased interest in the services of clinical sports psychologists. An appreciation of mind-body interactions and how they function regarding stress, sports performance, and injury is fundamental to the acceptance of psychological techniques in the medical arena. Teaching these fundamental issues to those in sports and medicine is essential. Furthermore, the psychology of sports injury needs continuing development of a base of theory, empirical research, and clinical practice that is sensitive to the needs of the individual athlete. Research on the assessment of psychosocial factors influencing sports injury and performance, as well as the efficacy of treatment modalities, is warranted. The psychology of sports injury has emerged from several previously established areas of psychology including behavioral medicine, rehabilitation, and sport psychology. As the techniques derived from these arenas are modified to suit the special needs of injured athletes, a set of principles and practices can be-established to better assist the sports medicine team in rehabilitation and prevention of sports injury. PMID- 9330814 TI - The significance of previous allergic contact dermatitis for elicitation of delayed hypersensitivity to nickel. AB - Several factors, such as amount of allergen, vehicle, anatomic site, immunologic status and previous eczema, may influence delayed hypersensitivity reactions. In an extended model, we have studied the significance of previous allergic contact dermatitis for elicitation of delayed hypersensitivity to nickel in 25 nickel allergic females. On 3 occasions, 8, 4 and 1 months before the final challenge patch testing, an experimental allergic contact dermatitis from nickel was induced on the lower back. At the challenge patch testing, 4 identical dilution series of nickel were tested on 4 areas on the lower back 3 with previous but healed dermatitis and 1 control area. The tests were read in a blind way. A significantly higher test reactivity was found at the areas with a previous allergic contact dermatitis, the shorter the time interval between the previous provocation and the challenge, the stronger the reaction. These results may be of importance for the understanding of factors contributing to chronicity of allergic contact dermatitis. PMID- 9330815 TI - Further evaluation of the quantitative structure-activity relationship for skin sensitizing alkyl transfer agents. AB - The biological activity of skin-sensitizing chemicals can be expressed in terms of physicochemical properties which relate to the propensity of those chemicals to behave as electrophiles and which describe their ability to partition into the epidermis and between compartments within it. For defined series of chemicals, it has proved possible to express such structure-activity relationships quantitatively. Such quantitative relationships can provide valuable insights into the mechanisms of skin sensitization and/or are of use in predictive toxicology. In the present work the quantitative structure-activity relationship (QSAR) previously derived for a series of alkyl transfer agents based on alkanesulfonate leaving groups has been critically examined in the light of skin sensitization data obtained for new members of that series and also for alkyl transfer agents based on different leaving groups. The QSAR predictions were broadly accurate, but demonstrated that further refinement was both necessary and possible. In particular, the physicochemical parameters which relate to the disposition of the chemical in the epidermis, i.e., its penetration through the stratum corneum, cell surface/cytoplasmic distribution and the associated dynamics, will need to be understood more fully in order to enhance the precision of the QSAR and its predictive power. PMID- 9330817 TI - More positive patch test reactions with larger test chambers? Results from a study group of the German Contact Dermatitis Research Group (DKG). AB - Test chambers of various sizes are commercially available for patch testing. Therefore, we asked the question whether the size of patch test chambers may affect allergic patch test reactions. A total of 495 patients were double tested synchronously with small and large Finn Chambers containing standard preparations of fragrance mix, wool wax alcohols, Kathon CG and formaldehyde. Double tests in 217 patients who had reacted with at least 1 allergic, questionable, or irritant reaction to 1 of these allergens were statistically evaluated. For each of the 4 allergens, a significantly higher number of stronger reactions was seen with the large chambers as compared to the small ones. It is concluded that large test chambers may be useful for detection of weak sensitizations to particular contact allergens. PMID- 9330816 TI - Nickel release from stainless steels. AB - In 1994, a study of nickel release and allergic contact dermatitis from nickel plated metals and stainless steels was published in this journal. It was shown that low-sulfur stainless steel grades like AISI 304, 316L or 430 (S < or = 0.007%) release less than 0.03 microgram/cm2/week of nickel in acid artificial sweat and elicit no reactions in patients already sensitized to nickel. In contrast, nickel-plated samples release around 100 micrograms/cm2/week of Ni and high-sulfur stainless steel (AISI 303-S approximately 0.3%) releases about 1.5 micrograms/cm2/week in this acid artificial sweat. Applied on patients sensitized to nickel, these metals elicit positive reactions in 96% and 14%, respectively, of the patients. The main conclusion was that low-sulfur stainless steels like AISI 304, 316L or 430, even when containing Ni, should not elicit nickel contact dermatitis, while metals having a mean corrosion resistance like a high-sulfur stainless steel (AISI 303) or nickel-plated steel should be avoided. The determining characteristic was in fact the corrosion resistance in chloride media, which, for stainless steels, is connected, among other factors, to the sulfur content. Thus, a question remained concerning the grades with an intermediate sulfur content, around 0.03%, which were not studied. They are the object of the study presented in this paper. 3 tests were performed: leaching experiments, dimethylglyoxime and HNO3 spot tests, and clinical patch tests; however, only stainless steels were tested: a low-sulfur AISI 304 and AISI 303 as references and 3 grades with a sulfur content around 0.03%: AISI 304L, AISI 304L added with Ca, AISI 304L+Cu. Leaching experiments showed that the 4 non resulfurised grades released less than 0.5 microgram/cm2/week in acid sweat while the reulfurized AISI 303 released around or more than 0.5 microgram/cm2/week. This is explained by the poorer corrosion resistance of the resulfurized grade. Yet all these grades had the same reaction to the DMG test (negative result), which shows again its lack of sensitivity. In contrast, the HNO3 spot test distinguished AISI 303 from the non-resulfurized grades. Clinical patch tests again showed that some patients (4%) were intolerant to AISI 303, while none were intolerant to the other grades. Thus, this study confirms that non-resulfurized stainless steels (S < or = 0.03%) like Ni-containing 304 and 304L should not elicit Ni contact dermatitis, while the resulfurized grades (S > 0.1%) should be avoided. PMID- 9330818 TI - Is it possible to improve the prognosis in nickel contact dermatitis? AB - A questionnaire was sent to 143 patients who had shown a positive patch test reaction to nickel sulfate more than 10 years earlier. 91 patients returned the questionnaire, revealing that after the testing, 73 had suffered from dermatitis, 61 especially from hand dermatitis. 37 of these patients were clinically examined and patch tested with standard series and in addition, 12 patients were tested with nickel sulfate and nickel chloride with different occlusion times. At the clinic visit, 23 patients had dermatitis, 16 hand dermatitis, and 11 were symptom free. 26 of the patients had metal items close to their skin and 21 of them had current dermatitis, 14 hand dermatitis. Of the 11 patients who had no metal exposure, 9 were symptom-free. The association of dermatitis with exposure to metal objects was statistically significant (p < 0.001). Those patients who had current dermatitis had also developed multiple allergies and reacted to nickel with shorter application times in patch tests, as compared to those who were symptom-free. It seemed possible that the prognosis for nickel dermatitis could be improved if nickel-allergic patients would strictly avoid metal contact, especially in clothing and jewelry. PMID- 9330819 TI - Contact urticaria due to rubber chemicals? AB - Contact urticaria due to gloves is mostly related to immediate-type allergy to natural rubber latex (NRL), and rarely due to rubber chemicals. We report the results of prick tests with rubber chemicals carried out on 75 latex-allergic patients. Clinically relevant hypersensitivity was not observed in our patient collection. Thus, we conclude that contact urticaria due to rubber chemicals is a very rare finding in latex-allergic patients, but has to be considered in patients with contact urticaria due to synthetic rubber gloves, as well as in patients with contact urticaria due to NRL gloves but no latex allergy. Therefore prick tests with rubber chemicals are unnecessary in the routine diagnostic assessment of latex-allergic patients. PMID- 9330820 TI - Changing clinical patterns of Parthenium dermatitis. PMID- 9330821 TI - Concomitant sensitization to Lannate and Gerbera. PMID- 9330822 TI - Photographic allergens: an update. PMID- 9330823 TI - Contact allergy to gold with pharyngeal and laryngeal disorders. PMID- 9330824 TI - Occupational dermatitis due to formaldehyde in newspaper. PMID- 9330825 TI - Allergic contact dermatitis from thiuram in a veterinary medication. PMID- 9330827 TI - Contact allergy to thiocolchicoside. PMID- 9330826 TI - Contact allergy to budesonide and perforation of the nasal septum. PMID- 9330828 TI - Occupational contact urticaria from paprika. PMID- 9330829 TI - Sensitivity to oxytetracycline. PMID- 9330830 TI - Allergic contact dermatitis from neticonazole hydrochloride. PMID- 9330831 TI - Non-occupational contact allergy to glutaraldehyde. PMID- 9330832 TI - Low-humidity dermatosis from car heaters. PMID- 9330833 TI - Dermatitis from chlorphenesin in a facial cosmetic. PMID- 9330834 TI - Photoallergic contact dermatitis due to flufenamic acid and etofenamate. PMID- 9330835 TI - Allergic contact urticaria from poppy flowers (Papaver rhoeas). PMID- 9330836 TI - Olive oil--contact sensitizer or irritant? PMID- 9330837 TI - Gastrointestinal decontamination after poisoning. Where is the science? AB - The approach to the use of gastrointestinal decontamination procedures in the treatment of ingested toxins has changed in recent years. Many toxicologists and physicians have taken strong positions either for or against the use of emesis, gastric lavage, activated charcoal, or other procedures. What is the scientific basis for these positions? This article reviews and comments on the published studies comparing the effectiveness of these widely used procedures. PMID- 9330838 TI - Anticonvulsant hypersensitivity syndrome. AB - Anticonvulsant hypersensitivity syndrome (AHS) is an uncommon but potentially fatal adverse effect that can occur from exposure to phenytoin, carbamazepine, or phenobarbital. It has diverse clinical features and a variable presentation which results in a delay in making the diagnosis. The syndrome commonly begins within 3 weeks after initiation of an anticonvulsant. Patients typically present with a constellation of fever, usually followed by the development of a rash of variable severity and type, and lymphadenopathy. In patients presenting with these features, the clinician should have a high index of suspicion for AHS. PMID- 9330839 TI - Antibiotic-induced convulsions. AB - Convulsive episodes are associated with the use of a number of antimicrobial agents. Although seizures may be a feature of the disease being treated, antibiotics should be considered possible causes of seizures, particularly if suggested by temporal relationships between seizure activity and drug administration. The astute clinician should be aware of the clinical settings in which antibiotic-induced seizures occur, be familiar with likely agents and their mechanisms of toxicity, and be prepared to institute appropriate management directed at this adverse effect of antimicrobial therapy. PMID- 9330840 TI - Serotonin syndrome. A clinical update. AB - Serotonin syndrome is characterized by varied degrees of cognitive, autonomic, and neuromuscular dysfunction and can only be produced by drug therapy that increases central nervous system serotonin neurotransmission. Information gained from a retrospective review of 127 cases of serotonin syndrome is presented. It is not uncommon for severe cases of serotonin syndrome to be confused with neuroleptic malignant syndrome. Treatment is mainly supportive, but specific pharmacologic therapy with serotonin antagonists may be potentially beneficial. PMID- 9330841 TI - Drug-induced hyperthermia. AB - Drug-related causes of hyperthermia can often be overlooked in the setting of elevated body temperature. This article reviews the pathophysiology, presentation, and treatment of several drug-induced hyperthermia syndromes: malignant hyperthermia, neuroleptic malignant syndrome, sympathomimetic poisoning, and anticholinergic toxicity. Although the general approach is similar, specific management strategies may be required for each syndrome. PMID- 9330842 TI - Evaluation of patients with chest pain after cocaine use. AB - Cocaine remains the most common cause of illicit drug-related visits to emergency departments, 40% of which result from chest pain. It is estimated that over half of the 64,000 patients evaluated annually for cocaine-associated chest pain will be admitted to hospitals for the evaluation of myocardial ischemia or infarction, at a health care cost of over eighty million dollars. Although the link between cocaine use and myocardial ischemia is well established, only about 6% of patients with cocaine-associated chest pain will demonstrate biochemical evidence of myocardial infarction. This article focuses on the evaluation of patients with chest pain following cocaine use, and concentrates on ways to improve diagnosis, management, and utilization of health care services. PMID- 9330843 TI - Poisoning by sodium channel blocking agents. AB - Poisoning by drugs that block voltage-gated sodium channels produces intraventricular conduction defects, myocardial depression, bradycardia, and ventricular arrhythmias. Human and animal reports suggest that hypertonic sodium bicarbonate may be effective therapy for numerous agents possessing sodium channel blocking properties, including cocaine, quinidine, procainamide, flecainide, mexiletine, bupivacaine, and others. PMID- 9330844 TI - Life-threatening plant poisoning. AB - Each year over 100,000 exposures to toxic plants are reported to poison control centers around the country. This article focuses on the more toxic plant exposures which may result in critical care admissions. The various plants are identified and described. Their mechanism of toxicity, clinical presentation of exposure and a management strategy for the critical care physician are discussed. Resources for further information are also listed. PMID- 9330845 TI - Crotalid snake envenomation. AB - Over 5000 Americans suffer from snake bites annually, and of these, nearly one quarter are from poisonous species. Although these cases are undeniably reported, death appears to occur in only a few cases each year, and often reflects delay in obtaining medical care. Two families of venomous snake indigenous to the United States account for most envenomations: Crotalidae (pit vipers or new world vipers) and Elapidae. This article focuses on the snakes of the Crotalidae family. PMID- 9330846 TI - Chemical warfare. Nerve agent poisoning. AB - The threat of civilian and military casualties from nerve agent exposure has become a greater concern over the past decade. After rapidly assessing that a nerve agent attack has occurred, emphasis must be placed on decontamination and protection of both rescuers and medical personnel from exposure. The medical system can become rapidly overwhelmed and strong emotional reactions can confuse the clinical picture. Initially, care should first be focused on supportive care, with emphasis toward aggressive airway maintenance and decontamination. Atropine should be titrated, with the goal of therapy being drying of secretions and the resolution of bronchoconstriction and bradycardia. Early administration of pralidoxime chloride maximizes antidotal efficacy. Benzodiazepines, in addition to atropine, should be administered if seizures develop. Early, aggressive medical therapy is the key to prevention of the morbidity and mortality associated with nerve agent poisoning. PMID- 9330847 TI - Hypericin inhibits choroidal endothelial cell proliferation and cord formation in vitro. AB - PURPOSE: To evaluate the effect of hypericin on bovine choroidal endothelial cell proliferation and cord formation and on protein kinase C activity. METHODS: The effect of hypericin (0.1-5 microM) on bovine choroidal endothelial cell proliferation was determined by cell number counting and a 3H-thymidine uptake assay in media containing 1, 5 or 10% serum. For the cord formation assay, bovine choroidal endothelial cells were seeded on basement membrane matrix, and the lengths of the capillary-like structures (cords) formed were quantified by image analysis. The effect of hypericin on cord formation was evaluated in the presence of serum or vascular endothelial growth factor. The effect of hypericin on protein kinase C activity was also measured in the presence or absence of light. RESULTS: Hypericin inhibited bovine choroidal endothelial cell proliferation in a dose-dependent manner in the presence of light but not in the dark. Serum dose dependently masked the inhibition of DNA synthesis by hypericin. Cord formation by bovine choroidal endothelial cells was stimulated by serum or vascular endothelial growth factor and inhibited by hypericin in the presence of light. Protein kinase C activity was completely inhibited by hypericin in the presence of light but only mildly inhibited in the absence of light. CONCLUSIONS: Hypericin inhibits bovine choroidal endothelial cell proliferation and cord formation and choroidal endothelial cell protein kinase C activity. These results suggest that hypericin should be further investigated in animal models for its potential to inhibit subretinal neovascularization. PMID- 9330848 TI - Human corneal epithelial cells reorient and migrate cathodally in a small applied electric field. AB - PURPOSE: To test whether human corneal epithelial cells (HCECs) respond to small applied electric fields (EFs) in a similar manner to bovine corneal epithelial cells (BCECs), the orientation and directed migration in small EFs of both primary cultures and of a human corneal epithelial cell line were quantified. METHODS: Primary cultures of human corneal epithelial cells (PHCECs) and transformed human corneal epithelial cells (THCECs) were exposed to EFs (100 mV/mm-250 mV/mm) in different media. Cell migration was traced using an image analyser. RESULTS: PHCECs and THCECs reoriented and migrated towards the cathode (negative pole) when cultured in small direct current (dc) EFs. Both the reorientation and directional migration were voltage- and serum-dependent, as shown previously for bovine cells. PHCECs and THCECs showed significant perpendicular orientation in EFs at 150 mV/mm in medium with serum, while at the same voltage, no significant orientation was found in serum free medium. PHCECs started to show perpendicular reorientation around 30 min after onset of EF at 150 mV/mm. They showed significant directional migration at 150 mV/mm, with directedness of 0.35 +/- 0.07 and a migration rate of 9.1 +/- 0.7 microns/h (n = 90), both significantly higher than that of cells in serum free medium. Addition of EGF-induced significant reorientation and directional migration of THCECs at 100 mV/mm. Additionally, as for BCECs, which remained viable and responsive to electric fields for at least 75 h at 150 mV/mm, THCECs also remained viable and showed responsiveness during long periods of exposure to EFs (at least 20 h). CONCLUSIONS: Cultured human primary CECs and a human corneal epithelial cell line both responded to small EFs with perpendicular reorientation and cathodally directed migration. Cell responses were qualitatively similar to those reported previously for bovine CECs. The endogenous EFs generated by wounded cornea may play an important role in promoting cell shape changes and directed migration of CECs during the healing process. PMID- 9330849 TI - Expression of hematopoietic cell markers by retinal pigment epithelial cells. AB - PURPOSE: We investigated the expression of various isoforms of the hematopoietic cell marker CD45 on retinal pigment epithelial cells in relation to their expression of CD68 and the cytokine-reactive intercellular adhesion molecule-1 (ICAM-1). We also determined the effect of the pro-inflammatory cytokines IL-1 beta, TNF alpha and IFN gamma on the expression of these molecules by RPE cells in culture. METHODS: Monolayers of RPE cells between 3rd and 7th passages were cultured in the presence or absence of cytokines, followed by immunohistochemical staining for CD45 (170-220 kD), CD45RA (205 and 220 kD), CD45RO (180 kD), CD68 and ICAM-1, using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique. Total (membrane and cytoplasmic) expression of each of the three CD45 isoforms was determined by enzyme-linked immunoassays (ELISA). RESULTS: The majority of RPE cells expressed all isoforms of CD45 on their membranes and the pattern of expression of these molecules was not modified by culture. The greatest intensity of membrane staining was consistently observed with antibodies to CD45RA (205 + 220 kD), while CD45 (170-220 kD) showed to be the predominant isoform within the whole cell, as judged by ELISA assays. Unlike the membrane expression of CD45, only 20% of RPE cells stained for the macrophage surface molecule CD68 following 4 h of culture, but progressive increase in the proportion of CD68 positive cells was observed by extending the culture to 24 and 48 h. Neither the expression of CD68 nor the various isoforms of CD45 were modified by incubation with pro-inflammatory cytokines. Staining for ICAM-1 was observed in 21-25% of RPE cells throughout the 48 h culture. However, incubation with 50 pg/ml of IL-1 beta, TNF alpha and IFN gamma caused a marked increase in the RPE cell expression of ICAM-1 following 4, 24 and 48 h culture. CONCLUSIONS: The observations suggest that hematopoietic cell markers are constitutively expressed on RPE cells and that functions governed by these molecules are not influenced by pro-inflammatory signals. Expression of hematopoietic molecules by RPE cells may influence the macrophage-like properties of these cells and may also aid in the identification of RPE cells during pathological processes, particularly in the proliferative retinopathies, where these cells undergo phenotypic and functional changes. PMID- 9330850 TI - The effect of nitric oxide synthase inhibitor on form-deprivation myopia. AB - PURPOSE: Form-deprivation myopia (FDM) is believed to result principally from actions of substances that modulate information processing in the retina. We used a chick model to investigate what role nitric oxide (NO), a gaseous neuromodulator, might play in the development of FDM. METHODS: We injected different concentrations of the NO synthase inhibitor N omega-nitro-L-arginine methyl ester (L-NAME) (30 ml) into the left eyes and the same volume of saline into the right eyes of 6-day-old chicks. Both eyes of most chicks were occluded for 6 days with translucent goggles. After removal of the goggles, the refraction was measured by retinoscopy and the axial lengths with an A-mode ultrasound. In some chicks we measured the concentration of NOx (nitrite and nitrate) in the retina. A few chicks, not wearing occluders after injection of L-NAME and saline, ERG and refraction, were examined 6 days after the treatment. RESULTS: In chicks that wore occluders, refractive error and axial length were significantly less affected in eyes injected with L-NAME (180, 360, or 540 mM) compared to control (right) eyes. ERG changes were reversible, except in eyes injected with the highest concentration (540 mM) of L-NAME. The eyes of chicks, injected with L NAME and reared without occlusion, had normal refractive values. After 6 days of form deprivation, the concentration of NOx in the retina of eyes injected with L NAME (180 mM) was significantly less than the concentration in eyes injected with saline. CONCLUSIONS: The injection of L-NAME before occlusion of developing chick eyes leads to reversible modifications in retinal function and inhibits the development of form-deprivation myopia. PMID- 9330851 TI - Free amino acids reflect impact of selenite-dependent stress on primary metabolism in rat lens. AB - PURPOSE: A decrease in phase separation temperature, prior to nuclear cataract, has been correlated with elevated free amino acid content. Hence, we determined how selenite-induced stress alters free amino acid pools in the rat lens, following a single subcutaneous dose of sodium selenite (30 nmol g-1 body weight) in 10- to 14-day-old Sprague Dawley rats. RESULT: Oxidative stress was evident in lenses 24 h after rats were treated with selenite. Glutathione content was decreased by 60% in the lens cortex and nucleus; the flux of glucose through the pentose phosphate pathway was increased; and glycerol-3-phosphate content was elevated. Amino acid transport, evaluated as 14C-cycloleucine uptake, was not altered, although 14C-glutamine was oxidized at a slower rate. Lenses from treated animals displayed, among the free amino acids, increased glutamine, proline, serine, glycine and the branched chain amino acids, while aspartate, glutamate, and taurine were less. CONCLUSIONS: A systemic delivery of sodium selenite caused oxidative stress in the rat lens. Direct effects on primary metabolism altered free amino acid pools that may contribute to transient and permanent changes in lens transparency. PMID- 9330852 TI - Mobilisation of intracellular calcium by P2Y2 receptors in cultured, non transformed bovine ciliary epithelial cells. AB - PURPOSE: To examine extracellular ATP for its ability to mobilise intracellular calcium in bovine ciliary epithelial cells; to establish and characterise P2Y2 receptor-mediated signal transduction in this tissue. METHODS: Bovine ciliary epithelial cells were isolated and cultured until confluence. The cells were reseeded on sterile coverslips and grown to obtain monolayers, then loaded with fura-2. Fluorescence was measured by a computer-controlled spectrofluorimeter and values calculated for intracellular calcium concentration. ATP, its analogues and other drugs were tested for their ability to mobilise intracellular calcium by adding them to the bathing solution. RESULTS: Basal cytosolic calcium in bovine ciliary epithelium was 138.4 +/- 0.8 nM (n = 274). In the presence of extracellular Ca2+, ATP, UTP or ADP induced a transient dose-dependent increase in intracellular calcium (maximum approx. 400%), which declined rapidly. The agonist potency order was UTP = ATP > ADP > AMP. Adenosine, alpha, beta-methylene ATP and 2-methylthio-ATP were ineffective in mobilising intracellular calcium, as were adrenaline, noradrenaline, acetylcholine and carbachol. The response to ATP and UTP remained, in the absence of extracellular calcium or the presence of nickel. Desensitisation of the calcium response by repeated exposure to ATP was augmented by phorbol-myristate-acetate and abolished by staurosporine. The ATP response was abolished by preincubation with pertussis toxin. Microfluorimetric measurements on single cells established that both pigmented and non-pigmented epithelia responded to ATP or UTP similarly. CONCLUSIONS: In the bovine ciliary epithelium, ATP stimulates P2Y2 receptors coupled to a pertussis toxin-sensitive G protein. The results also suggest that this receptor activates phospholipase C, leading to mobilisation of calcium from intracellular stores. PMID- 9330853 TI - Zinc induces catalase expression in cultured fetal human retinal pigment epithelial cells. AB - PURPOSE: We have previously shown that an experimental, low-zinc environment decreased catalase activity in cultured human fetal retinal pigment epithelial (RPE) cells. The purpose of this study was to investigate the effect of zinc supplementation on catalase expression in cultured human fetal RPE cells. METHODS: Confluent fetal RPE cells incubated in Coon's modified Ham's F12 (CMF 12) were treated (18 h) with zinc chloride (ZnCl2) (15, 30, or 100 microM) to assess changes in catalase enzyme activity or for 6 h to assess the induction of catalase mRNA by Northern analysis and in situ hybridization. RPE cells were also treated with 30 microM ZnCl2 for 2, 6, 24, 48 and 72 h to assess the time course of changes in catalase enzyme activity, changes in mRNA levels and status of the Sp1 transcription factor. RESULTS: Catalase activity was increased above control by the addition of 15, 30 and 100 microM ZnCl2. Catalase gene expression was induced by 30 microM zinc in 6 h, but decreased to non-treated control levels by 24 h. The transcription factor Sp1 was also activated by zinc treatment (30 microM) which peaked at 2 h and declined to non-treated control levels by 24 h. Catalase enzyme activity peaked at 24 h and decreased to control levels by 72 h. CONCLUSIONS: Our results demonstrate that zinc treatment of RPE cells increases catalase expression and activates the transcription factor Sp1. The results suggest zinc may play a role in the transcriptional regulation of catalase in RPE cells. PMID- 9330854 TI - Effects of changes in intraocular pressure on human ocular haemodynamics. AB - PURPOSE: Myogenic autoregulation is the ability of a vascular bed to maintain blood flow despite changes in perfusion pressure. Ocular perfusion pressure is defined as the difference between ocular arterial pressure and ocular venous pressure, the latter dependent on intraocular pressure (IOP). The aim of the present study was to investigate the effect of moderate increases in IOP on ocular haemodynamics. METHODS: Changes in IOP (+ 10 mmHg, +20 mmHg) were induced by a suction cup in 10 healthy subjects. Ocular fundus pulsations in the macula and the optic disc were measured by laser interferometry; blood flow velocities in the central retinal artery (CRA) and in the ophthalmic artery (OA) were measured by Doppler sonography. RESULTS: Changes in IOP caused a significant reduction in fundus pulsations, which was more pronounced in the macula (at +10 mmHg: -9 +/- 2%, p < 0.01; at +20 mmHg: -19 +/- 3%, p < 0.001) than in the optic disc (at +10 mmHg: -5 +/- 2% (ns); at +20 mmHg: -9 +/- 3%, p < 0.01). Mean flow velocity in the CRA was reduced by -5 +/- 3% at +10 mmHg (ns) and by -14 +/- 5% at +20 mmHg (p < 0.005), resistive index was increased by +4 +/- 1% at +10 mmHg (p < 0.05) and by +6 +/- 2% at +20 mmHg (p < 0.01). In contrast, a rise in IOP did not affect blood flow parameters in the OA. CONCLUSIONS: Our results from fundus pulsation measurements indicate that choroidal blood flow decreases when IOP is increased. The Doppler sonographic findings in the CRA indicate reduced blood flow velocity in this artery during raised IOP. PMID- 9330855 TI - Antiproliferative effect of intravitreal alpha-tocopherol and alpha-tocopheryl acid-succinate in a rabbit model of PVR. AB - PURPOSE: To evaluate the antiproliferative properties of alpha-tocopherol and alpha-tocopheryl-acid-succinate in a rabbit model of proliferative vitreoretinopathy. METHODS: Fifty-seven rabbits underwent gas-compression vitrectomy and gas/fluid exchange. Group 1 (n = 8): 50 micrograms alpha tocopherol in 1 ml of 0.5% ethanol in balanced salt solution; Group 2 (n = 8): 50 micrograms alpha-tocopheryl-acid-succinate in 1 ml of 0.5% ethanol in balanced salt solution; Groups 3 (n = 4) and 4 (n = 2): 1 ml 0.5% ethanol in balanced salt solution; Groups 5 (n = 12) and 8 (n = 9): alpha-tocopherol in 1 ml silicone oil (12 mg/ml); Group 6 (n = 9): 1 ml silicone oil; Group 7 (n = 5): 1 ml balanced salt solution. Groups 1-3, and 5-7 also received fibroblasts and platelet-rich plasma injection. Fundus evaluation was performed during a four-week period. The eyes were enucleated for gross examination on day 28. Histopathology was performed on Group 4. RESULTS: Alpha-tocopherol and alpha-tocopheryl-acid succinate in saline solution delayed development of proliferative vitreoretinopathy, compared to the control group (statistically significant during the first week, Mann Whitney, p < 0.05). The alpha-tocopherol in the silicone-oil group delayed development of proliferative vitreoretinopathy, compared to the silicone-oil group during the second to fourth weeks (no statistically significant difference, p > 0.05). CONCLUSIONS: Alpha-tocopherol and alpha-tocopheryl-acid-succinate in saline solution showed retardation of proliferative vitreoretinopathy traction retinal detachments. In silicone oil, alpha-tocopherol is slowly released and decreases the severity of proliferative vitreoretinopathy, especially during the second week of follow-up. PMID- 9330857 TI - Grating detection and orientation discrimination in amblyopia. AB - PURPOSE: We examined whether the misperceptions associated with amblyopic visual perception can be revealed under natural viewing conditions by comparing the ability to detect the presence of a grating with the ability to identify the grating orientation. METHODS: Grating detection and orientation discrimination performance (horizontal versus vertical) were determined, using stimuli that consisted of sinusoidal gratings of fixed contrast (75%) but with variable spatial frequency. A total of four amblyopic subjects (two strabismic and two non strabismic) and four age-matched normals participated in the experiment. RESULTS: Psychometric functions for grating detection and orientation identification were found to be closely matched in the normal subjects and in all four amblyopic subjects, indicating that orientation could be correctly identified at detection threshold. CONCLUSIONS: The absence of orientation uncertainty in the psychophysical data for the amblyopic observers is not consistent with the several previous reports of spatial aliasing in the central field of amblyopes. Our results suggest that non veridical visual perception in central amblyopic vision can not be revealed under natural viewing conditions by comparing the ability to detect the presence of a grating with the ability to identify its orientation. Possible reasons for the failure of this technique to reveal spatial aliasing in amblyopes are discussed. PMID- 9330856 TI - Immunopathologic features of Staphylococcus epidermidis-induced endophthalmitis in the rat. AB - PURPOSE: To investigate the clinical, histopathologic and immunologic responses to Staphylococcus epidermidis endophthalmitis in a rat model. METHODS: Experimental rats received an intravitreal injection of viable S. epidermidis (7000 organisms), while control rats received sterile saline. The clinical scores, cellular infiltrate in vitreous, and levels of serum and vitreous IgM, IgG and IgA to glycerol teichoic acid (GTA), the major antigenic determinant of S. epidermidis cell wall, were all measured from day 1 to day 30 after injection. RESULTS: The ocular inflammation was largely resolved by day 14. The red reflex was abolished in 50% of rats between days 3 and 10. The bacteria were cleared from the vitreous by day 7. In vitreous, the neutrophils peaked at day 1 and decreased by day 7, and plasma cells were seen between days 1 and 3. Presence of B cells (CD45+/CD3-) was confirmed by flow cytometric analysis of pooled vitreous humor. IgM and IgG but not IgA antibodies to GTA were found in vitreous of injected eyes. The peak of anti-GTA IgM was observed in vitreous of S. epidermidis-infected rats on day 1 and declined by day 7. In contrast to vitreous antibodies, serum anti-GTA IgM antibodies were significantly elevated throughout the course of S. epidermidis endophthalmitis. A weak IgG but no IgA response were observed in serum. Anti-GTA antibodies were also found in low level in normal sera but not in normal vitreous. CONCLUSIONS: The vitreous antibodies may be involved in neutrophil-mediated opsonophagocytosis leading to 'spontaneous sterility' of the bacteria, and may play a role in the immunopathogenesis of staphylococcal endophthalmitis in the rat. PMID- 9330858 TI - Kinetic evidence for Na(+)-glucose co-transport in the pigmented rabbit conjunctiva. AB - PURPOSE: To obtain kinetic evidence for the existence of a Na(+)-coupled glucose co-transport process on the mucosal (tear) side of the pigmented rabbit conjunctiva. METHODS: The excised pigmented rabbit conjunctiva was mounted in the modified Ussing chamber for measurement of 22Na and 3H-3-O-methyl-D-glucose (3-O MG) fluxes. RESULTS: In the presence of 5 mM glucose, the conjunctival tissue showed net Na+ absorption in the mucosal-to-serosal direction at an approximate rate of 0.15 microEq/cm2/h. This net Na+ absorption was abolished by serosally added 0.5 mM ouabain, but not affected by mucosally added 0.1 mM or 1 mM amiloride. There was a 40-60% reduction in net Na+ absorption under the glucose free condition or in the mucosal presence of 0.5 mM phlorizin. Moreover, serosally added ouabain and mucosally added phlorizin (both at 0.5 mM) significantly decreased the 3-O-MG permeability coefficient in the mucosal-to serosal direction by about 70%, whereas mucosally instilled 0.1-1.0 mM amiloride was without any effect. Three-O-MG absorption in the mucosal-to-serosal direction appeared to be coupled with Na+ transport with a 1:1 stoichiometry. In addition, this process exhibited temperature dependency, saturability, and directionality. CONCLUSION: Our findings are consistent with Na(+)-glucose cotransport as being one of the mechanisms for mucosal Na+ entry into the epithelial cells of the pigmented rabbit conjunctiva. PMID- 9330859 TI - The antimicrobial susceptibility of Mycobacterium chelonae isolated from corneal ulcer. AB - PURPOSE: To determine the in vitro susceptibility of Mycobacterium chelonae isolates from corneal ulcers to various traditional and newly-developed antimicrobial agents, alone or in combination. METHODS: Fifteen strains of M. chelonae isolated from corneal ulcers were collected at the National Taiwan University Hospital from 1989 to 1993. Susceptibility to antimicrobial agents was tested by the broth microdilution method to determine the minimum inhibitory concentration (MIC). The antimicrobial effects of combinations of antimicrobial agents were assessed by the checkerboard titration method to determine the fractional inhibitory concentration (FIC) index. RESULTS: The MIC results showed that traditional antituberculous drugs had poor activity against M. chelonae. In the aminoglycoside group, tobramycin and amikacin had better activity than gentamicin. Among macrolides, clarithromycin was especially effective, with an MIC ranging from 0.125 to 1 microgram/ml. Among various beta-lactam antibiotics, imipenem was the only one to demonstrate good anti-mycobacterial activity. Of the quinolone group, ciprofloxacin was the most effective, with an MIC ranging from 0.5 to 16 micrograms/ml. Combination of an aminoglycoside with imipenem, ciprofloxacin or clarithromycin all showed antagonistic effect. CONCLUSIONS: The results suggested that amikacin, clarithromyicn, imipenem and ciprofloxacin had good in vitro antimicrobial activity against M. chelonae. However, no synergistic effect could be demonstrated for combinations of an aminoglycoside with other effective drugs. PMID- 9330860 TI - Repeated topical administration of fenoterol in rabbit reverses its initial ocular hypotensive effect and decreases sensitivity of adenylyl cyclase in ciliary processes to stimulatory agents. AB - PURPOSE: The effects of repeated topical administration of the selective beta 2 adrenergic agonist fenoterol on the intraocular pressure and on the adenylyl cyclase activity in ciliary processes in rabbit were examined in order to detect their possible causal relationship. METHODS: Intraocular pressure was measured by pneumatonometry. Adenylyl cyclase activity in homogenates of ciliary processes was assayed ex vivo by measurement of conversion of 32P-alpha-ATP to 32P-cyclic AMP. RESULTS: A single topical dose of 1% solution of fenoterol elicited a clear cut decrease of the intraocular pressure lasting for several h. Repeated administration of fenoterol for 2-5 days led to a significant increase of intraocular pressure, observable from the second to the fifth day. The stimulation of adenylyl cyclase activity ex vivo by isoproterenol, vasoactive intestinal polypeptide or forskolin was significantly decreased on the fifth day (24 h after the administration of the last dose of fenoterol). CONCLUSIONS: Our data showed that repeated topical administration of the selective beta 2 adrenergic agonist increased intraocular pressure and desensitized adenylyl cyclase in ciliary processes; if these two effects are related then they would support the idea of direct relationship of decreased cAMP production in ciliary processes to the increase of intraocular pressure, and vice versa. However, conclusive evidence of this suggestion and of its possible significance in another animal species or man would require further study. PMID- 9330861 TI - An immunohistochemical study of TNF-alpha in optic nerves from AIDS patients. AB - PURPOSE: Both in vitro and in vivo studies have implicated a role for tumor necrosis factor (TNF-alpha) in the pathology of demyelinating diseases. The purpose of this study was to address the hypothesis that TNF-alpha is a mediator of AIDS-related optic nerve injury and to determine the cell types involved in the proliferation of TNF-alpha in the AIDS optic nerve. METHODS: Ten optic nerves from seven patients with AIDS, and three from persons who were HIV negative were stained, using the indirect immunoperoxidase method. Six of the ten AIDS optic nerves were positive for cytomegalovirus (CMV), but the remainder did not have abnormal fundus findings. RESULTS: In all the optic nerves from AIDS patients with or without CMV retinitis, the vast majority of astrocytes stained strongly for TNF-alpha. Microglial cells (MPS-derived macrophages) varied from not staining to staining strongly positive for TNF-alpha. However, oligodendrocytes were not labeled positively for TNF-alpha. Some endothelial cells also stained for TNF-alpha. Examination of normal optic nerves and controls did not reveal any cell type that stained positively for TNF-alpha. CONCLUSIONS: The present study supports the contention that TNF-alpha is a major mediator of AIDS-associated optic neuropathy. HIV infection induces the production of TNF-alpha in macrophages and astrocytes, which probably causes demyelination and other neuronal damage. PMID- 9330862 TI - Aqueous hyaluronic acid concentration: comparison in pediatric and adult patients. AB - PURPOSE: To determine if there is an age-related increase in human aqueous hyaluronic acid (HA) concentration. METHODS: HA concentrations were measured in 102 specimens of human aqueous humour obtained during intraocular surgery. Patient age ranged from one month to 93 years. Measurement of the HA concentration in the specimens was performed by a modified ELISA-like assay using a biotinylated HA-binding peptide. RESULTS: An approximate five-fold increase in the mean aqueous HA concentration was observed between the pediatric (0.33 microgram/ml, n = 5) and the adult patients (1.72 micrograms/ml, n = 97, p < 0.0002). Among the adult patients, however, there was a poor correlation between age and aqueous HA concentration. CONCLUSIONS: Adult aqueous humor has a significantly higher HA concentration than aqueous obtained from pediatric patients. The source of this increase is unclear, but may be from anterior segment production of HA, or alternatively, from anterior diffusion of vitreous HA. PMID- 9330863 TI - The Eph family of receptors. AB - Eph receptor tyrosine kinases have recently been identified as instructive molecules that guide the topographic movement of cells and growth cones. The activation of Eph receptors by their ligands, which are membrane-anchored molecules, involves a cell-cell recognition event that often causes cell repulsion. Therefore, Eph receptors mediate signals that can override cell adhesion. Transmembrane ligands for Eph receptors also exhibit properties of signal transducing molecules, suggesting that bidirectional signaling occurs when receptor-expressing cells contact ligand-expressing cells. PMID- 9330864 TI - CEA adhesion molecules: multifunctional proteins with signal-regulatory properties. AB - The carcinoembryonic antigen family comprises a large number of complex molecules, several of which possess cell adhesion activities. The primordial adhesion molecules of this family are the cell-cell adhesion molecules (C-CAMs), which have been found to be multifunctional, signal-regulatory proteins. C-CAMs inhibit tumor growth, interact with calmodulin, protein tyrosine kinases and protein tyrosine phosphatases, and are subject to specific dimerization reactions. These new insights indicate that C-CAMs are important regulators of cellular functions. PMID- 9330865 TI - Neural recognition molecules and synaptic plasticity. AB - Recent studies of neural recognition molecules have revealed similarities between their functions during ontogenetic development and in neural plasticity in the adult. Observations both at the cellular level in vitro and at the behavioural level in vivo suggest that altered recognition molecule expression can lead to changes in synaptic efficacy, and alterations in synaptic function in turn evoke changes in recognition molecule expression. These changes can manifest themselves as morphological alterations and modulations of the synapse's signal transduction machinery. PMID- 9330867 TI - Leukocyte adhesion: CD11/CD18 integrins and intercellular adhesion molecules. AB - Leukocyte integrins and intercellular adhesion molecules play pivotal roles in leukocyte adhesion to target cells and extracellular matrices. Recently, novel intercellular adhesion molecules have been identified, and much information has been obtained on the structures and binding sites of leukocyte integrins and of intercellular adhesion molecules. Furthermore, much progress has been made in the study of integrin activation and the role of leukocyte adhesion molecules in disease. PMID- 9330866 TI - The importance of cellular environment to function of the CD44 matrix receptor. AB - Much has been learned recently by experimental manipulation of the structure of CD44 and assessment of the resulting functions. However, even greater structural variation is naturally introduced by CD44-bearing cells. A structural model is now available for the portion of CD44 that recognizes hyaluronan, but it is clear that all domains of the molecule influence CD44 functions. PMID- 9330868 TI - The dermal-epidermal junction. AB - Recent insights into the structure and function of the dermal-epidermal junction have resulted from two converging lines of experimental evidence, namely, the study of inherited blistering disorders of the skin, in which mutations in genes encoding proteins of this region have been discovered, and the targeted ablation of the same genes in knockout mouse models. In addition to these studies, elegant analyses of the cell biology of the hemidesmosome/anchoring filament complex have revealed not only functionally important interactions between structural protein components, but also the role of certain of these proteins in mediating cell adhesion, migration, and signal transduction of messages from the extracellular matrix into the keratinocyte. Our current understanding of the dermal-epidermal junction forms a new model encapsulating the nature both of the hemidesmosomal attachment structures and of the interhemidesmosomal attachments that are mediated by differential cell type specific expression of proteins of the cutaneous adhesion zone. PMID- 9330869 TI - The ezrin protein family: membrane-cytoskeleton interactions and disease associations. AB - Ezrin, radixin, moesin and merlin form a subfamily of conserved proteins in the band 4.1 superfamily. Ezrin protein subfamily members act as linkers between the plasma membrane and the cytoskeleton. Members of the subfamily have been shown to interact with each other, with cell adhesion molecules such as CD44 and with F actin. Recent data indicate that intercellular adhesion molecules 1 and 2 also interact with ezrin. The proteins are also involved in the redistribution of intercellular adhesion molecules and the organization of cell membrane structures. Merlin is a tumor suppressor that is involved in tumorigenesis of schwannomas and meningiomas. Merlin has the same overall protein structure as the other proteins in the subfamily but may have partially distinct functions. PMID- 9330870 TI - Natural killer cell-target cell interactions. AB - Research into the molecular mechanisms of target cell recognition by natural killer (NK) cells has recently progressed rapidly. NK cells express several MHC I recognizing receptors that inactivate the NK cells' functions. In pathological alterations of MHC I expression, the NK cell inhibitory receptors do not engage and thus permit the lysis of the target cell. The receptors that trigger the cytolytic machinery of NK cells, after the permission of lysis from the inhibitory receptors, are poorly characterized. Some candidate triggering receptors have been identified and it seems that triggering of NK cell killing is mediated by multiple receptors, as is the inhibition of cytotoxicity. PMID- 9330871 TI - Interendothelial junctions: structure, signalling and functional roles. AB - Endothelial cell-cell adhesive junctions are formed by transmembrane adhesive proteins linked to a complex cytoskeletal network. These structures are important not only for maintaining adhesion between endothelial cells and, as a consequence, for the control of vascular permeability, but also for intracellular signalling properties. The establishment of intercellular junctions might affect the endothelial functional phenotype by the downregulation or upregulation of endothelial-specific activities. PMID- 9330872 TI - Cadherins, catenins and APC protein: interplay between cytoskeletal complexes and signaling pathways. AB - Cadherins play important roles in cell-cell adhesion during tissue differentiation. Cadherins are linked to the actin cytoskeleton by catenins (beta catenin/armadillo, plakoglobin, and alpha-catenin). Recent results show that beta catenin also binds to another cytoskeletal complex containing the adenomatous polyposis coli protein and microtubules, and interacts with several signaling pathways that include tyrosine kinases and phosphatases and Wnt/Wingless. Interplay between these cytoskeletal complexes and signaling pathways may regulate morphogenesis. PMID- 9330873 TI - Integrin signaling: specificity and control of cell survival and cell cycle progression. AB - Integrin-mediated adhesion to the extracellular matrix plays an important role in regulating cell survival and proliferation. There is now increasing evidence that integrins activate shared as well as subgroup-specific signaling pathways. The signals from these adhesion receptors are integrated with those originating from growth factor and cytokine receptors in order to organize the cytoskeleton, stimulate mitogen-activated protein kinase cascades, and regulate immediate early gene expression. The repertoire of integrins and composition of the extracellular matrix appear to dictate whether a cell will survive, proliferate or exit the cell cycle and differentiate in response to soluble factors. PMID- 9330874 TI - Integrins and anoikis. AB - The loss of integrin-mediated cell-matrix contact induces apoptosis ('anoikis') in certain cell types. Recently it has been shown that protein kinase signaling pathways control anoikis both positively and negatively. Focal adhesion kinase, when activated by integrins, can suppress anoikis. Phosphatidylinositol 3-kinase and the AKT oncoprotein may mediate the anoikis-suppressing effects of focal adhesion kinase. Conversely, the stress-activated protein kinase/Jun amino terminal kinase pathway promotes anoikis. Latest results indicate that caspase mediated cleavage of the first component of this latter pathway, MEKK-1, may trigger activation of this pathway in anoikis. In addition, certain integrins may regulate bcl-2 expression levels, possibly adjusting the threshold for anoikis. PMID- 9330875 TI - Role of integrins in cellular responses to mechanical stress and adhesion. AB - Mechanical stresses are important environmental cues for both normal cellular functions and pathophysiological changes in conditions such as cardiac hypertrophy and atherosclerosis. There is increasing evidence that mechanotransduction processes in response to mechanical stresses share many common features with processes in cell adhesion, such as an increase in tyrosine phosphorylation of proteins in the focal adhesion sites. Recent findings suggest that integrins may function as mechanotransducers in cells. PMID- 9330877 TI - Integrins and inside-out signal transduction: converging signals from PKC and PIP3. AB - Recent studies have identified molecules that interact with integrins and appear to participate in the signaling pathways that regulate integrin adhesiveness. Clues provided by studies of these molecules point to the integration by integrins of signal transduction pathways implicated in cell division and activation. PMID- 9330876 TI - Plasminogen activators, integrins, and the coordinated regulation of cell adhesion and migration. AB - Cellular migration is critically dependent on an interplay between forces of attachment and detachment. Recent studies show that the serine protease urokinase and its major inhibitor and receptor regulate the adhesive properties of integrins, at least in part through initiation of cellular signals. These new functions for an old protease system imply intricate connections between proteolysis and adhesion that operate at the cell surface to regulate migration. PMID- 9330878 TI - Cell-to-cell contact and extracellular matrix. PMID- 9330879 TI - Guillain-Barre syndrome: an evolving concept. PMID- 9330881 TI - Neuromuscular disorders in systemic malignancy. AB - Paraneoplastic neuronopathies are presumed to be the result of an autoimmune attack directed at neuronal proteins, and both humoral and cell-mediated mechanisms have been postulated. The lower motor neuron syndrome after irradiation to the spinal column is caused by a proximal motor polyradiculopathy. Prevention of brachial plexopathy after radiotherapy for breast cancer may be accomplished by lower doses and surgical management of the axilla. Polymerase chain reaction casts doubt on the distinction between neoplastic and paraneoplastic mechanisms of neuromuscular manifestations of lymphoproliferative diseases. Shared antigenic components may underlie the association between inflammatory neuropathy and malignant melanoma. Advances in chemotherapy strategies against responsive tumors are hindered by the toxic effects of agents on peripheral nerves. PMID- 9330880 TI - Autoimmune ataxic neuropathies (sensory ganglionopathies). AB - Autoimmune ataxic neuropathies are a subset of the sensory ataxic neuropathies which are characterized by ataxia as the dominant presenting feature. The major known causes of autoimmune ataxic neuropathies include sensory variants of the Guillain-Barre syndrome, including Miller-Fisher syndrome, subsets of immunoglobulin M paraproteinaemic neuropathy, paraneoplastic neuropathy and the neuropathy associated with Sjogren's syndrome. Identified antigens as targets for autoantibodies include gangliosides, myelin associated glycoprotein, Hu antigen and extractable nuclear antigens. Some recent studies support the pathogenic role of anti-GD1b ganglioside antibody in autoimmune ataxic neuropathies. The major site of pathology in autoimmune ataxic neuropathies is the dorsal root ganglion, but dorsal roots and peripheral nerve myelin and axons may also be affected. PMID- 9330882 TI - Diabetes mellitus. AB - Epidemiological studies have documented a high prevalence of diabetic neuropathy. The risk of lower leg amputation is increased three to four times in patients with clinical signs of neuropathy and ankle weakness is more common than hitherto recognized. Increased nerve hydration and increased expression of low affinity p75 receptor for neurotrophins suggest new therapeutic potentials for the prevention of diabetic neuropathy. PMID- 9330883 TI - Spinal muscular atrophy. AB - Proximal childhood spinal muscular atrophy is a common autosomal recessive disorder that results in degeneration of lower motor neurons of the spinal cord. The defective gene, survival of motor neuron, encodes a novel protein with a putative role in RNA metabolism. Further work is required to define clearly the mechanism by which the survival of motor neuron gene defect would result in motor neuron degeneration. PMID- 9330884 TI - The biology and pathobiology of Schwann cells. AB - The most common forms of inherited demyelinating neuropathy in humans are caused by mutations in the genes encoding protein zero, peripheral myelin protein 22 kDa, and connexin32, all of which are expressed by myelinating Schwann cells and are components of the myelin sheath. The phenotype of myelinating Schwann cells depends on the maintenance of axon-Schwann cell interactions, because axonal degeneration also leads to the breakdown of the myelin sheath and dedifferentiation of the previously myelinating cells into 'denervated' Schwann cells, which are essential for axonal regeneration. Several transcription factors have been shown to play critical roles in regulating the phenotype of Schwann cells, including SCIP/tst-1/Oct-6 and Krox-20, both of which are required for the normal development of the myelinating phenotype. PMID- 9330885 TI - Myotonic dystrophy--forgotten aspects of an often neglected condition. PMID- 9330886 TI - Congenital myasthenic syndromes. AB - Congenital myasthenic syndromes are a rare group of heterogeneous disorders affecting neuromuscular transmission. Recent identification and in-vitro functional analysis of some of the genetic mutations that cause these disorders correlates with previous electrophysiological, biochemical, pathological and therapeutic studies, and has advanced our understanding of neuromuscular transmission. PMID- 9330887 TI - Genetic counselling in mitochondrial diseases. AB - Mitochondrial disorders may be caused by mutations either in mitochondrial or in nuclear genes involved in the synthesis or regulation of respiratory chain subunits. The unique nature of the mitochondrial genome calls for a different approach to genetic counselling and risk analysis. PMID- 9330888 TI - Inclusion body myositis and myopathies. AB - Sporadic inclusion body myositis is a frequent, acquired, adult-onset vacuolar myopathy affecting proximal and distal muscles with a distinct, easily identifiable clinical pattern. Although its primary cause is still unknown, autoimmune, viral, and degenerative processes, alone or in combination, are being considered. A uniform and sustained therapeutic response using the currently available immunomodulatory agents has not yet been achieved. Hereditary, inherited noninflammatory rimmed vacuolar myopathies with similar histologic features, collectively called hereditary inclusion body myopathies, are being redefined with the use of molecular genetics. The implications of the recent advances in clinical and basic sciences are discussed in the present review. PMID- 9330889 TI - Emery-Dreifuss syndrome. AB - Emery-Dreifuss syndrome is a heterogeneous entity characterized by the following clinical triad: early contracture of the elbows. Achilles tendons and postcervical muscles; slowly progressive muscle wasting and weakness with a humeroperoneal distribution early in the course of disease; and a cardiomyopathy usually presenting as an atrioventricular block ranging from sinus bradycardia to complete heart block. As the heart block is the major problem, insertion of a cardiac pacemaker can be life saving. Recent advances through genetic and immunochemical studies have provided valuable clues to the understanding and the early diagnosis of this disease. PMID- 9330890 TI - Desmin-related myopathies. AB - Desmin-related myopathies are marked by accumulation of desmin, which is often familial and associated with cardiomyopathy. When multifocal this excess is characterized by inclusions such as cytoplasmic or spheroid bodies, when disseminated the excess is called granulofilamentous material. Excess of desmin might represent an abnormal type of protein metabolism. PMID- 9330891 TI - Gene therapy research for Duchenne and Becker muscular dystrophies. AB - Gene therapy is a promising option for the definitive treatment of Duchenne and Becker muscular dystrophies. Presently, gene therapy for Duchenne and Becker muscular dystrophies is still in the preclinical stage with dystrophin-deficient animals (the mdx mouse and a golden retriever dog strain) serving as convenient models. The thrust of research during the past 18 months has focused on two approaches: adenovirus-mediated dystrophin gene transfer and upregulation of a natural dystrophin analogue, utrophin. In the area of adenovirus-mediated gene transfer, substantial progress has been made in characterizing and mitigating the deleterious immune responses to the vector and transgene proteins. Furthermore, new adenovirus vectors have been created with reduced immunogenicity and increased insert gene capacity, which enhance the longevity of the transgene expression. Additional efforts are underway to develop safe and efficient routes of administration of the adenovirus vector carrying the dystrophin expression cassette. The prospects of utrophin upregulation as an attractive strategy for treatment of Duchenne and Becker muscular dystrophies was greatly enhanced by the demonstration of a substantial mitigation of the dystrophic phenotype of the transgenic mdx mouse overexpressing utrophin. PMID- 9330893 TI - Neuromuscular diseases: nerve. PMID- 9330892 TI - Functions of dystrophin and dystrophin associated proteins. AB - Dystrophin is a protein product of the X-linked gene mutation that is responsible for Duchenne and Becker muscular dystrophies. The protein binds actin and associates with dystrophin-glycoprotein complex to link the cytoskeleton to the extracellular matrix. Defects in the components of the dystrophin-glycoprotein complex are responsible for several phenotypes of muscular dystrophy. PMID- 9330894 TI - Neuromuscular diseases: muscle. PMID- 9330895 TI - The reliability of the items of the Functional Assessment Measure (FAM): differences in abstractness between FAM items. AB - The reliability of the Functional Assessment Measure (FIM+FAM) is an important issue with its increased use in the measurement of neurological disability and rehabilitation outcome. Although the Motor items have good reliability ratings, the Cognitive items are more difficult to complete and their reliability is not as good. This study tests the suggestion that this might be due to the Cognitive items being more abstract. A keyword from each of four Motor items was compared with a keyword from four Cognitive items. Abstractness was measured by measuring the 'imageability' of each keyword. The Motor items were found to have a significantly higher mean imageability rating than the Cognitive items. Thus, there is support for the suggestion that abstractness contributes to the poorer reliability of the Cognitive items. These results led to the proposal that the reliability of the Cognitive items might be improved by various methods of increasing the tangibility of these measures (e.g. subdivision of broad categories of disabilities, enhancing item descriptions, training raters to increase their recognition of relevant observations, and using specific assessment tasks to elicit relevant behaviours). PMID- 9330896 TI - Rehabilitation in the Mediterranean Basin--towards the next millennium. PMID- 9330897 TI - The current status of rehabilitation in Spain. PMID- 9330898 TI - Physical medicine and rehabilitation in France. PMID- 9330899 TI - Rehabilitation in Italy. PMID- 9330900 TI - The state of rehabilitation in Slovenia. PMID- 9330901 TI - The status of rehabilitation in Turkey. PMID- 9330902 TI - Rehabilitation medicine in Israel. PMID- 9330903 TI - Rehabilitation in Belgium. PMID- 9330904 TI - Rehabilitation in the United Kingdom. PMID- 9330905 TI - Prediction of translation initiation sites on the genome of Synechocystis sp. strain PCC6803 by Hidden Markov model. AB - We developed a computer program, GeneHackerTL, which predicts the most probable translation initiation site for a given nucleotide sequence. The program requires that information be extracted from the nucleotide sequence data surrounding the translation initiation sites according to the framework of the Hidden Markov Model. Since the translation initiation sites of 72 highly abundant proteins have already been assigned on the genome of Synechocystis sp. strain PCC6803 by amino terminal analysis, we extracted necessary information for GeneHackerTL from the nucleotide sequence data. The prediction rate of the GeneHackerTL for these proteins was estimated to be 86.1%. We then used GeneHackerTL for prediction of the translation initiation sites of 24 other proteins, of which the initiation sites were not assigned experimentally, because of the lack of a potential initiation codon at the amino-terminal position. For 20 out of the 24 proteins, the initiation sites were predicted in the upstream of their amino-terminal positions. According to this assignment, the processed regions represent a typical feature of signal peptides. We could also predict multiple translation initiation sites for a particular gene for which at least two initiation sites were experimentally detected. This program would be effective for the prediction of translation initiation sites of other proteins, not only in this species but also in other prokaryotes as well. PMID- 9330906 TI - Differences in dinucleotide frequencies of human, yeast, and Escherichia coli genes. AB - Nucleotide sequences coding proteins in human, yeast and Escherichia coli genes were analyzed in terms of dinucleotide occurrences. Every gene is plotted as a point in the dinucleotide space, which is spanned by 16 axes corresponding to the 16 components of the dinucleotide. The metric unit in the space is defined using the log-odds ratio of dinucleotide occurrences in a gene. The distribution of points showed that genes from the same organism are clustered in the space. The clusters of human and E. coli are completely separated, and the yeast cluster sits between, implying that individual genes are classified into the three sources from their location. In fact, they could be identified with accuracy of 90%, using the DNA data alone. Even genes encoding homologous proteins belonging to the same protein superfamily were discriminated by the DNA data, and were correctly identified into their sources with the same accuracy as above. DNA sequences of non-coding regions, including human introns, as well as human genes of GC-rich and GC-poor types, were also analyzed in the same manner. The most significant finding is that human genomic DNA sequences, including genes and introns together, exhibit the largest deviation of dinucleotide occurrence from the random expectation. Possible origins for this phenomenon are discussed. PMID- 9330907 TI - Rolling-circle plasmid pKYM re-initiates DNA replication. AB - It is believed that rolling-circle plasmids are incapable of re-initiation since they have to maintain their copy number and this is one of the differences between plasmids and phages as phi-x174. To examine whether a rolling-circle plasmid pKYM is incapable of re-initiating DNA replication, we constructed a plasmid that carries both the pKYM origin (fragment 13, 173 bp) and its truncated origin (fragment 32, 56 bp) in the same orientation. This plasmid yielded two smaller plasmids in the presence of RepK, an initiator protein. We showed that RepK can bind to the fragment 13 but not to fragment 32 which lacks the 3'- moiety of fragment 13. These results imply that RepK initiates DNA replication from fragment 13 and terminates at fragment 32, then the same RepK is used for re initiation of replication from the fragment 32 region. pKYM is likely to be a unique plasmid that re-initiates DNA replication like a phase phi-x174. PMID- 9330908 TI - Synthesis of RepK of rolling circle plasmid pKYM is regulated by countertranscript and HU protein. AB - Replication of rolling circle plasmid pKYM was regulated by RepK, a plasmid encoded initiator protein, with HU protein and antisense RNA (copRNA) that block the expression of RepK. HU protein bound to the repK promoter in the presence of RepK protein and inhibited the transcription of repK mRNA. The copRNA would hybridize to repK mRNA and induce a stem-loop structure in which the repK Shine Dalgarno sequence is sequestered by base pairing. Sequence substitution experiments demonstrated that this stem-loop not only inhibits translation but induces premature termination. PMID- 9330909 TI - Identification of likely genes on chromosome VI of Saccharomyces cerevisiae by correlating transcripts and nucleotide sequence data. AB - Most of the 97 transcripts of the genes on chromosome VI of Saccharomyces cerevisiae that were identified by a series of Northern hybridization experiments (Yoshikawa and Isono, Nucl. Acids Res., 19, 1189-1195, 1991) have been correlated with the open reading frames (ORFs) deduced from the nucleotide sequence data of this chromosome (Murakami et al., Nature Genet., 10, 261-268, 1995). This was performed by comparing the experimentally constructed physical map and the one produced from the nucleotide sequence data, as well as the sizes and positions of observed transcripts and those of sequenced ORFs. Thus, 75 ORFs of chromosome VI were correlated uniquely with the corresponding transcripts and 3 ORFs with two transcripts of different sizes. Comparing the relative abundance levels of individual transcripts with that of the RPO41 transcript, highly expressed genes of chromosome VI were found to be located almost exclusively on the Crick strand. Based on the correlation between the abundance level of the experimentally identified transcripts and the codon adaptation indices of the corresponding gene, the genes on chromosome VI of S. cerevisiae were classified into three groups. The data thus provides information concerning their chromosomal locations as well as their likely levels of expression in vegetatively growing cells. PMID- 9330910 TI - Structural analysis of Arabidopsis thaliana chromosome 5. I. Sequence features of the 1.6 Mb regions covered by twenty physically assigned P1 clones. AB - A total of 20 P1 clones with an average insert size of 80 kb and each containing a marker(s) specifically mapped on chromosome 5 were isolated from a P1 library of the Arabidopsis thaliana genome, and their nucleotide sequences were determined according to a shotgun-based strategy and precisely located on the physical map of chromosome 5 separately constructed. The total length of the sequenced regions were summed up to 1,621,245 bp. By comparison with the sequences in protein and EST databases and analysis with computer programs for gene modeling, a total of 347 potential protein-coding genes and/or gene segments with known or predicted functions were identified. The positions of exons which do not exhibit any similarity to known genes were also predicted. An average density of the genes and/or gene segments assigned so far as 1 gene/4,672 bp. Introns were identified in approximately 78% of the potential genes, and the average number and length of the introns per gene were 3.7 and 161 bp. The transcription level of the predicted genes was roughly monitored by counting the numbers of identified Arabidopsis ESTs. The sequence data and gene information are available through the World Wide Web at http:/(/)www.kazusa.or.jp/arabi/. PMID- 9330911 TI - Evaluation of cDNA libraries from different developmental stages of Schistosoma mansoni for production of expressed sequence tags (ESTs). AB - A comparative study of the gene expression profile in different developmental stages of Schistosoma mansoni has been initiated based on the expressed sequence tag (EST) approach. A total of 1401 ESTs were generated from seven different cDNA libraries constructed from four distinct stages of the parasite life cycle. The libraries were first evaluated for their quality for a large-scale cDNA sequencing program. Most of them were shown to have less than 20% useless clones and more than 50% new genes. The redundancy of each library was also analyzed, showing that one adult worm cDNA library was composed of a small number of highly frequent genes. When comparing ESTs from distinct libraries, we could detect that most genes were present only in a single library, but others were expressed in more than one developmental stage and may represent housekeeping genes in the parasite. When considering only once the genes present in more than one library, a total of 466 unique genes were obtained, corresponding to 427 new S. mansoni genes. From the total of unique genes, 20.2% were identified based on homology with genes from other organisms, 8.3% matched S. mansoni characterized genes and 71.5% represent unknown genes. PMID- 9330912 TI - Difference in HMG1-induced DNA bending among microsatellites. AB - Sequence-dependency of high-mobility group protein (HMG) 1-induced DNA bending is examined for microsatellites using a circularization assay which can measure the extent of bending. Fragments of 133 bp containing (GGA/TCC)11 in the middle showed greater bending than those harboring (GAA/TTC)11 and (GT/AC)17 repeats, and fragments possessing (GA/TC)17 exhibited only slight bending. Differences were not detected for fragments having the repeats near the end. Filter binding assays showed no difference in their binding affinity, suggesting that GGA/TCC repeats are more flexible than the other three repeats as concerns HMG1-induced bending. These results suggest that the mammalian genomes comprise flexible and inflexible regions of microsatellites which might play roles in chromatin architectures and in dynamic packaging of genomic DNA during the cell division cycle. PMID- 9330913 TI - Physical mapping of human 7q and 14q subtelomeric DNA sequences in the great apes. AB - Phylogenetic divergence of the members of the Pongidae family has been based on genetic evidence. The terminal repeat array (T2AG3) has lately been considered as an additional basis to analyze genomes of highly related species. The recent isolation of subtelomeric DNA probes specific for human (HSA) chromosomes 7q and 14q has prompted us to cross-hybridize them to the chromosomes of the chimpanzee (PTR), gorilla (GGO) and orangutan (PPY) to search for its equivalent locations in the great ape species. Both probes hybridized to the equivalent telomeric sites of the long (q) arms of all three great ape species. Hybridization signals to the 7q subtelomeric DNA sequence probe were observed at the telomeres of HSA 7q, PTR 6q, GGO 6q and PPY 10q, while hybridization signals to the 14q subtelomeric DNA sequence probe were observed at the telomeres of HSA 14q, PTR 15q, GGO 18q and PPY 15q. No hybridization signals to the chromosome 7-specific alpha satellite DNA probe on the centromeric regions of the ape chromosomes were observed. Our observations demonstrate sequence homology of the subtelomeric repeat families D7S427 and D14S308 in the ape chromosomes. An analogous number of subtelomeric repeat units exists in these chromosomes and has been preserved through the course of differentiation of the hominoid species. Our investigation also suggests a difference in the number of alpha satellite DNA repeat units in the equivalent ape chromosomes, possibly derived from interchromosomal transfers and subsequent amplification of ancestral alpha satellite sequences. PMID- 9330914 TI - An arrayed human not I-EcoRV boundary library as a tool for RLGS spot analysis. PMID- 9330915 TI - Collecting substance use data with an anonymous mailed survey. AB - Because mailed surveys minimize personal contact, they are useful for collecting sensitive data on substance use, as long as the problems of achieving adequate response rates can be conquered. To address these issues, we report on an anonymous mailed survey of substance use with a 78% response rate, including data collection and survey methods. Analysis of sociodemographic effects on responding found certain groups required additional contacts. Substance use estimates were not affected by non-response bias, suggesting that anonymous mailed surveys can be a feasible means of collecting data on substance use. PMID- 9330916 TI - Subjective sleep-wake parameters in treatment-seeking opiate addicts. AB - We investigated subjective sleep parameters and sleep difficulties of opiate addicts undertaking methadone detoxification and identified their sleep profile. Using the St Mary's Sleep Questionnaire, we compared the subjective sleep parameters of 27 consecutively consenting patients (16 males, 11 females) with a mean age of 33 years (S.D. = 7.5) undertaking in-patient methadone detoxification with those of 26 drug-free controls (9 males, 17 females) with a mean age of 35 years (S.D. = 8.0). Our findings reveal that subjective sleep parameters of opiate addicts and controls are quantitatively and qualitatively different. The patients are more likely than controls to report difficulty initiating sleep (OR = 5.42; 95% CI = 1.43, 20.47); difficulty maintaining sleep (OR = 16.50; 95% CI = 3.81, 71.47); inadequate sleep quality (OR = 8.56; 95% CI = 2.04, 35.81); and inadequate sleep quantity (OR = 9.00; 95% CI = 2.49, 32.57). PMID- 9330917 TI - Ethanol alters hemodynamic responses to cocaine in rats. AB - Cocaine is often used while consuming ethanol despite evidence that this combination may enhance the toxicity of cocaine. In the present study, we examined the cardiovascular effects of ethanol (475 or 950 mg/kg, i.v.) alone and in combination with cocaine (5 mg/kg, i.v.) in conscious rats. Ethanol or cocaine administration produced a consistent pressor response but highly variable cardiac output and systemic vascular resistance responses. The hemodynamic response patterns in individual rats to either drug were similar and related within rats. After ethanol pretreatment, cocaine produced greater decreases in cardiac output. We have proposed that this pattern of responses may reflect a predisposition in individual rats to cocaine-induced cardiomyopathies and hypertension. Furthermore, these data suggest that ethanol administration elicits a similar pattern of hemodynamic responses as previously reported for cocaine or amphetamine administration or acute behavioral stress. PMID- 9330918 TI - Imagery of craving in opiate addicts undergoing detoxification. AB - Craving is a significant factor in opiate addiction that is associated with drug dependence and in relapse to drug use after treatment. In order to better understand the psychological and physiological mechanisms of craving for opiates, we have developed an imagery-based procedure using personal verbal descriptions of craving in abstinent opiate addicts. Thirteen opiate addicts in detoxification were required to imagine and describe their craving experiences while autonomic measures of heart rate and arterial pressure were taken. Subjects displayed a significant increase in systolic blood pressure and heart rate while describing drug craving compared with neutral descriptions. Furthermore, an increase in systolic blood pressure during imagery of craving descriptions compared with neutral descriptions was observed. These results provide preliminary evidence that imagery is powerful in eliciting craving for opiates, as indicated by subjective ratings and autonomic measures. The implications of the results of this paper for the cue-exposure paradigm and contemporary models of addiction are being discussed. PMID- 9330919 TI - Attendance incentives for outpatient treatment: effects in methadone- and nonmethadone-maintained pregnant drug dependent women. AB - The effectiveness of behavioral incentives for improving treatment participation and retention in samples of methadone-maintained (n = 66) and nonmethadone maintained (n = 76) pregnant drug dependent women was examined. Subjects were randomly assigned to receive $0 (standard care) and $1, $5, or $10/day for attending at least 4 h of interdisciplinary treatment programming during the first 7 consecutive days after transfer from residential to outpatient care, with payment dispensed in the form of gift certificates. Methadone-maintained women attended nearly twice as many full treatment days as those not receiving methadone (5.2 vs 2.8 days; P < 0.001) and were retained in treatment significantly longer (86.4 vs 28.9% active in treatment at 30 days). There was no main effect of incentives and no effect on attendance in methadone patients. However, nonmethadone patients offered higher magnitude incentives ($5/$10) attended 3.3 days out of 7 on average, compared to 2.3 days for those offered $0 or $1 per day (t = 1.73; P < 0.05). The study confirmed that methadone maintenance is a powerful therapeutic adjunct which is associated with significantly better treatment retention and participation in ancillary programming than is abstinence-based treatment. It was also found that modest financial incentives can facilitate treatment participation for abstinence-based patients. However, more potent interventions would be needed to match the effectiveness of methadone in this regard. PMID- 9330920 TI - Use of identical assay conditions for cocaine analog binding and dopamine uptake to identify potential cocaine antagonists. AB - The addictive and euphorogenic properties of cocaine are thought to result from inhibition of the dopamine transporter (DAT). Recent evidence suggests that dopamine and cocaine bind to distinct sites on the transporter protein. Therefore it should be possible to design drugs which specifically inhibit cocaine recognition by the DAT while permitting the transporter to maintain its function of accumulating dopamine. One way to monitor such activity is to compare the inhibition constants of test agents for inhibition of radiolabelled dopamine uptake (Kiuptake) and inhibition of the binding of a cocaine ligand such as [3H]2 beta-carbomethoxy-3 beta-3 beta-(fluorophenyl)tropane (CFT; Kibind) and select for compounds with Kiuptake/Kibind ratios greater than unity. Because others have shown that compounds can exhibit Kiuptake/Kibind ratios greater than unity when the assays are performed under non-identical conditions, we have established these assays under identical conditions of time, temperature and buffer using a Chinese hamster ovary (CHO) cell line which stably expresses the human DAT. Kinetic and saturation analyses were performed on both assay and over 200 structurally diverse compounds were screened. Using identical assay parameters, several series of compounds having Kiuptake/Kibind ratios significantly greater than unity were identified. Such compounds include local anesthetics (procaine, dibucaine, tolperisone, dyclonine, diperodone), antipsychotic agents (10 (diethylaminopropionyl)phenothiazine), antidepressants (desipramine, imipramine, protriptyline), a diuretic (5-N-methyl-N-isobutyl-amilioride), an anticholinergic agent (prindinol), a PKC inhibitor (H-8), a calcium channel antagonist (loperamide) and an antimalarial compound (chloroquine). To our knowledge, even though these compounds exhibit low binding affinities (3-24 microM), they represent some of the most cocaine site-selective compounds identified to date using identical assay parameters. PMID- 9330921 TI - Assessing the helping alliance and its impact in the treatment of opiate dependence. AB - This study assesses the relationship between the patient-counselor helping alliance (HA) and progress in methadone maintenance treatment. Questionnaire measures of HA were administered to 57 patients 1 and 3 months after admission. Three-month HA measures (especially counselors' ratings) predicted reductions in drug use as measured by weekly urinalysis results and 6-month self-report data. HA was unrelated to treatment retention or improvement in psychiatric symptomatology. Moreover, controlling for urinalysis results in the previous month rendered insignificant the correlations between 3-month HA and subsequent drug use. Thus, this evaluation of the HA's unique contribution to the prediction of outcome suggests that the development of a positive HA may be more a marker of treatment progress than a necessary precursor of positive outcomes in the methadone maintenance treatment setting. PMID- 9330922 TI - Methodology for clinical trials in rheumatology: logical foundations. PMID- 9330923 TI - Standardized patients: uses beyond the undergraduate years. PMID- 9330924 TI - Real standardized arthritis patients are better. Let me tell you how. PMID- 9330925 TI - Is Behcet's disease part of the spondyloarthritis complex? PMID- 9330926 TI - Alternatively spliced CS-1 fibronectin isoform and its receptor VLA-4 in rheumatoid arthritis synovium. AB - OBJECTIVE: Extracellular matrix components and cell adhesion molecules play a role in the pathogenesis of rheumatoid arthritis (RA). Interaction between the integrin very late antigen-4 (VLA-4) and the connecting segment-1 (CS-1) fibronectin (FN) isoform may contribute to lymphocyte interaction in RA synovium. We examined both mRNA and protein expression of CS-1 FN in inflamed synovium, and VLA-4 expression in synovial tissue and on cultured fibroblast-like synoviocytes from patients with RA. METHODS: Snap frozen synovial tissue specimens of 10 patients with RA and 4 patients with osteoarthritis were examined for expression of CS-1 FN mRNA and protein by in situ hybridization and immunohistochemistry. VLA-4 expression of synovial fibroblasts and in synovial tissue was evaluated by flow cytometry and immunohistochemistry. RESULTS: CS-1 FN mRNA was detected in RA lining layer synoviocytes, around terminal vessels, and in endothelial cells. Double labeling revealed that most lining synoviocytes expressing CS-1 FN mRNA were CD68 negative. VLA-4 was found in RA synovial fibroblasts, sublining mononuclear cells, and lymphoid aggregates. CONCLUSION: Our findings suggest that expression of CS-1 FN may partially correlate with cell proliferation in the RA lining layer. VLA-4 was found in RA synovial lining, as well as on cultured synovial fibroblasts. Thus, VLA-4/CS-1 FN interaction may facilitate lymphocyte interaction and synovial proliferation in RA. PMID- 9330927 TI - Long-term treatment of destructive rheumatoid arthritis with methotrexate. AB - OBJECTIVE: To evaluate the tolerability and efficacy of methotrexate (MTX) treatment in patients with longstanding, progressive, active rheumatoid arthritis (RA) who had failed one or more disease modifying antirheumatic drugs (DMARD). METHODS: Two hundred seventy-one consecutive patients with RA in whom MTX treatment was introduced were followed at regular intervals for up to 108 months. Evaluations included the number of swollen joints, grip strength, patient assessment of pain and mobility, erythrocyte sedimentation rate (ESR), and hemoglobin. Radiographs of hands and feet were taken once a year and 32 joints were evaluated according to a modified Larsen score. RESULTS: Of the 271 patients, 269 had prior treatment with one DMARD, primarily parenteral gold, and 58% with 2 or more DMARD. MTX was started parenterally in all patients in doses between 15 and 25 mg weekly and continued by oral medication in most of the cases. Eighty-three percent of patients complained of adverse events. The most common side effects were nausea, hair loss, transaminase increase, and stomatitis. In 45 patients (16.5%), MTX was withdrawn because of side effects, mostly during the first year. Sixteen patients (5.9%) died during followup, mainly due to myocardial infarction, heart failure, stroke, or cancer. After one year, 78.7% and after 5 years 60.3% of the patients were still taking MTX. Number of swollen joints, ESR, grip strength, patient assessment of pain, and mobility improved significantly at all measurement points. Improvement in the swollen joint count and the ESR of over 50% was seen in more than 50% of patients. Inactivation of the disease, defined as < 2 swollen joints, ESR < 20 mm, and no concomitant steroid use, occurred in 8-14% of patients. Steroid intake was significantly reduced. In spite of clinical improvement the modified Larsen score showed a progression in the vast majority of patients. CONCLUSION: Even in patients with longstanding, active, destructive RA who failed one or more DMARD, MTX treatment is well tolerated and improves clinical and biochemical disease activity significantly, while radiographic progression is still present. PMID- 9330928 TI - Short-term low dose methotrexate ameliorates abnormal bone metabolism and bone loss in adjuvant induced arthritis. AB - OBJECTIVE: To clarify whether short term weekly methotrexate (MTX) therapy aggravates bone abnormalities in adjuvant induced arthritis (AIA) or improves them through its antiarthritic effect. METHODS: Bone metabolism and bone mineral density (BMD) were studied in 6 groups of Lewis rats: (1) normal controls, (2) rats given MTX 0.3 mg/kg weekly, (3) rats given MTX 3 mg/kg weekly, (4) AIA rats, (5) AIA rats given MTX 0.3 mg/kg weekly, and (6) AIA rats given MTX 3 mg/kg weekly. Osteogenic activity was determined from serum osteocalcin levels and number of marrow fibroblast colony forming units (osteogenic precursor cells). Bone resorptive activity was assayed by detecting osteoclast-like cells and pit formation in bone marrow cultures. RESULTS: In control rats, MTX (3 mg/kg weekly) suppressed osteogenic activity, as shown by low serum osteocalcin levels and decreased growth of marrow fibroblast colony forming units. Osteoclast-like cells and pit formation in bone marrow cultures from control rats were increased by MTX, but BMD was unchanged. In rats with AIA, MTX (3 mg/kg) suppressed arthritis and restored the decreased osteogenic activity of bone marrow cells, and reduced their increased bone resorptive activity. These changes resulted in a significant increase of periarticular BMD in the femur. CONCLUSION: Low dose weekly MTX therapy had a favorable effect on abnormal bone metabolism and osteopenia in rats with AIA. PMID- 9330929 TI - Safety and efficacy of hydroxychloroquine as maintenance therapy for rheumatoid arthritis after combination therapy with methotrexate and hydroxychloroquine. AB - OBJECTIVE: To evaluate the ability of hydroxychloroquine sulfate (HCQ) to extend the response to combination therapy with HCQ and methotrexate (MTX) and the safety of longterm HCQ maintenance therapy in patients with active rheumatoid arthritis (RA). METHODS: Two-part study consisting of an open label segment evaluating combination HCQ/MTX therapy followed by a double blind segment evaluating maintenance therapy for a total of 60 weeks. First, all patients were treated with HCQ 400 mg/day and MTX 7.5 to 15 mg/week for 24 weeks. Then, responders were randomized into 3 groups: (1) HCQ with MTX as needed for disease flare (n = 40), (2) HCQ 400 mg/day (n = 41), or (3) placebo with MTX as needed for disease flare (n = 40), each for 36 weeks. RESULTS: Clinical disease and laboratory variables improved significantly during initial combination therapy with HCQ and MTX. After MTX withdrawal, HCQ-containing maintenance regimens delayed the onset of disease flare (p = 0.023). There were no unexpected adverse events at any time or between-group differences in the distribution of adverse events during the double blind segment. CONCLUSION: Combination of HCQ and MTX appeared to be effective and well tolerated for 24 weeks. After withdrawal of MTX, HCQ extended the response seen with combination therapy and was well tolerated for 36 weeks. Initial therapy with HCQ and MTX, followed by maintenance HCQ, may be a useful alternative for the treatment of RA. PMID- 9330930 TI - Total and free methotrexate pharmacokinetics in elderly patients with rheumatoid arthritis. A comparison with young patients. AB - OBJECTIVE: To determine the pharmacokinetics of methotrexate (MTX) in a group of patients 65 to 83 years of age and to compare the pharmacokinetic data to those in patients 21 to 45 years of age. METHODS: Thirty-eight elderly patients (8 men, 30 women) and 24 young patients (6 men, 18 women) underwent this pharmacokinetic study. They received intramuscular administration of MTX each week, at a dose varying from 7.5 to 15 mg depending on the patient. MTX concentrations in plasma and ultrafiltrate samples were assayed by a fluorescence polarization immunoassay. Pharmacokinetic variables were estimated using a Bayesian approach with population parameters as a priori information together with 2 plasma MTX concentrations (2 and 8 h after injection). RESULTS: The extent of unbound fraction and the volume of distribution were not statistically significantly different between the 2 age groups. The elimination half-life measures of the free and total MTX were greatest in the elderly group (p < 0.001). The total clearances of free and total MTX were inversely proportional to age (p < 0.001). CONCLUSION: MTX clearance decreases with decreasing creatinine clearance and smaller doses may be chosen in this group. Thus, a dosage regimen should be adjusted in elderly patients or in those with renal impairment. PMID- 9330931 TI - A cross sectional assessment of health status instruments in patients with rheumatoid arthritis participating in a clinical trial. Minocycline in Rheumatoid Arthritis Trial Group. AB - OBJECTIVE: To (1) validate the Short-Form Health Survey (SF-36) as a generic functional health status measure in patients with rheumatoid arthritis (RA); and (2) assess correlations between the SF-36 and other outcome measures used in the Minocycline in Rheumatoid Arthritis (MIRA) Trial. METHODS: We conducted a cross sectional analysis of the final visit outcome measures from the 48 week, multicenter, placebo controlled, double blind MIRA trial. Multitrait scaling analyses assessed convergent and discriminant validity and internal consistency reliability of the SF-36 in the study patients. Responses to comparable items on the SF-36 and modified Health Assessment Questionnaire (M-HAQ) regarding physical functioning were compared and questions from both instruments were also compared to other RA outcome measures. RESULTS: In patients with RA, the SF-36 had high internal consistency and reliability, high discriminant and high convergent validity. Moderate correlations were observed (r = -0.46 to -0.61, p < 0.01 in each case) for comparable items on the SF-36 and M-HAQ regarding dressing, walking, and bending. Joint tenderness score correlations with items on the M-HAQ and SF-36, and joint tenderness score correlations with the SF-36 scales were higher than for joint swelling scores. Physician and patient global assessments were most highly correlated (r = 0.58 and 0.66; p < 0.01, respectively) with the SF-36 bodily pain item. CONCLUSION: The SF-36 is a valid instrument for this RA population. The SF-36 correlates with the M-HAQ and the physician and patient global assessments. The usefulness of the SF-36 in measuring change in RA clinical trials requires testing in longitudinal studies. PMID- 9330932 TI - Bone scintigraphy of the hands in early stage lupus erythematosus and rheumatoid arthritis. AB - OBJECTIVE: To evaluate retrospectively the discriminatory value of bone scintigraphy, especially spot images of the hands, in differentiating early stage systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). METHODS: Data from 19 patients with SLE (3 men, 16 women) and 20 patients with RA (6 men, 14 women), presenting with early stage articular disease (arbitrarily defined as articular complaints for no longer than 3 mo), were reviewed. At this stage, radiographs were normal in all patients. In all 39 patients, total body bone scintigraphy with spot images of the hands was performed as part of a complete diagnostic investigation. For differentiation between SLE and RA in early disease stage, less extensive semiquantitative description in 3 categories (normal, diffuse mildly increased, and (multi)focal moderately to markedly increased tracer accumulation) proved to be sufficient. Locations of bone scintigraphic findings were correlated to clinical findings. RESULTS: In RA, bone scintigraphy revealed foci of moderate to markedly increased tracer accumulation, corresponding to the sites of clinical synovitis in all patients. In 10 patients with SLE, bone scintigraphy images of the hands were normal, and in 9 patients diffuse mildly increased tracer accumulation was observed. CONCLUSION: The data suggest bone scintigraphy may be useful to differentiate SLE from RA in early stage disease. PMID- 9330933 TI - Prevalence of reduced bone mineral density in systemic lupus erythematosus and the role of steroids. AB - OBJECTIVE: To determine the prevalence of reduced bone mineral density (BMD) in a large female cohort of systemic lupus erythematosus (SLE) and to determine the role of steroids and disease related variables. METHODS: All females with SLE managed by rheumatologists affiliated with a single center were invited to undergo BMD measurement of the lumbar spine and left femoral neck by dual energy X-ray absorptiometry (DEXA), standardized examination, and medical record review. RESULTS: Ninety-seven females with a mean (SD) age of 44.2 (14.9) years were studied. Low bone mass [defined as BMD > 1 standard deviation (SD) below young adult mean] was present in 44.3 and 42.1% at the lumbar spine and femoral neck, respectively. Osteoporosis (defined as BMD > 2.5 SD below young adult mean) was present in 13.4 and 6.3% at the lumbar spine and femoral neck, respectively. Steroid usage showed a strong inverse relationship with BMD in the lumbar spine, but a less strong relationship in the femoral neck. CONCLUSION: The findings of high prevalence of reduced BMD and association with steroid therapy have important implications for the routine management of SLE. PMID- 9330934 TI - Epidermal growth factor, transforming growth factor-alpha, and epidermal growth factor receptor in labial salivary glands in Sjogren's syndrome. AB - OBJECTIVE: Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) affect cells through binding to a shared EGF receptor (EGF-R), which is a transmembrane protein with tyrosine kinase activity. They exert trophic effects on vascular endothelial, salivary acinar, and ductal and mucosal epithelial cells. In Sjogren's syndrome (SS) focal sialadenitis leads to salivary gland tissue damage, diminished salivary flow, and changes in the oral epithelium, a complex referred to as xerostomia. We compared the localization of EGF, TGF-alpha, and EGF-R in labial salivary glands in SS and in healthy controls. METHODS: Labial salivary gland tissues of 12 patients with SS and 7 healthy controls were stained with the immunohistochemical peroxidase antiperoxidase method for EGF, TGF-alpha, and EGF-R. RESULTS: Immunoreactivity for both EGF and TGF-alpha was found in endothelial cells of blood vessels and in some ductal epithelial cells. TGF-alpha, but not EGF, was also found in some acinar cells. EGF-R was found in endothelial, acinar, and salivary duct epithelial cells. There was no difference in the expression of EGF-R between diseased and healthy specimens, but both EGF and TGF-alpha were diminished in SS. CONCLUSION: The interrelated localization of EGF-R and its ligands, EGF and TGF alpha, suggests an autocrine, juxtacrine, and paracrine mitogenic/trophic role for them and thus a role in the maintenance of the secretory and excretory cells of the normal salivary glands. The trophic effects on acinar cells seem not to be mediated by EGF, but more likely by TGF-alpha. The diminished expression of EGF and TGF-alpha indicates a failure of this trophic system in SS, which may contribute to the acinar atrophy and secondary changes thereof, including atrophy of the oral mucosa. PMID- 9330935 TI - In situ expression and serum levels of tumor necrosis factor-alpha receptors in patients with early stages of systemic sclerosis. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) is an important cytokine in the early stage of systemic sclerosis (SSc), which is characterized by mononuclear cell infiltration and microvascular alterations. Most effects of TNF alpha are mediated by its interaction with 2 types of TNF receptors and depend on their surface expression on individual cell subsets. Our purpose was to correlate the serum levels of soluble TNF receptors-TNF-RI(p55) and RII(p75)-with (1) their in situ expression and distribution in lesional skin and on peripheral blood mononuclear cells (PBMC), and (2) the clinical disease progression and inflammatory serum variables in patients with SSc. METHODS: Serum samples of 32 patients with SSc and 36 healthy probands were examined by ELISA. We performed immunohistological stainings and in situ hybridization on cryostat sections of skin lesions, cytometric analysis on PBMC, and reverse transcriptase polymerase chain reactions using RNA from cultured skin fibroblasts in 17 of these 36 patients. RESULTS: In contrast to healthy skin and chronic fibrotic SSc, TNF-RI is expressed on about 30% of mononuclear infiltrating cells in early skin lesions. Neither TNF-RI nor RII was detectable on fibroblasts by immunohistochemistry, but specific mRNA could be found on the transcriptional level. TNF-RII is found on most lymphocytes and on 30-50% of endothelial cells, especially in early SSc. Expression of both receptor types on PBMC in patients and controls was not significantly different. Serum levels of soluble TNF-RI and RII correlated well with their in situ expression and with clinical and laboratory signs of inflammation and disease progression in patients with SSc. CONCLUSION: Our data provide evidence for a central role of the TNF-alpha/TNF-R system in the early pathological events of scleroderma with prominent inflammation and endothelial cell damage. Determination of TNF-R serum levels provides a useful diagnostic tool for characterization of the disease stage and progression, and to guide experimental therapy in patients with SSc. PMID- 9330936 TI - Effect of iloprost infusion on the resistance index of renal vessels of patients with systemic sclerosis. AB - OBJECTIVE: To investigate the effect of iloprost, a stable prostacycline analog, on kidney blood flow in patients with systemic sclerosis (SSc), using color flow Doppler sonography. METHODS: The acute effect of the drug was studied in 10 patients with SSc with elevated resistance index (RI) levels (all RI values reported are multiplied by 100). Iloprost was administered intravenously (2 ng/kg/min for a period of 8 h). To study the effects of chronic drug administration, 16 patients with SSc were randomly assigned to 2 groups of 8 cases each. The first group was treated with 9 infusions of iloprost in 6 mo. The second group was treated with slow release nifedipine (40 mg/day) for 6 mo. RESULTS: Interlobar artery RI (median 67 vs 61; p = 0.02) and cortical vessel RI (median 65 vs 54; p = 0.001) were reduced after acute treatment. In chronic drug administration, RI values were not modified by nifedipine, while iloprost reduced the RI of the interlobar (median 69 vs 61; p < 0.03) and cortical arteries (median 66 vs 58: p < 0.01). CONCLUSION: Our findings suggest iloprost might be useful for treatment of scleroderma renal vasospasm. PMID- 9330937 TI - Patterns of psoriatic arthritis in Orientals. AB - OBJECTIVE: To determine the clinical features of psoriatic arthritis (PsA) in a multiethnic Oriental population and to study the effect of ethnicity on disease patterns. METHODS: A retrospective study of 80 patients with PsA seen at either a rheumatology or dermatology referral center. Patients and case records were reviewed and data abstracted according to a standard protocol. Eighty consecutive patients with psoriasis without PsA seen at the dermatology center were recruited as controls. RESULTS: Asymmetric polyarthritis developing in the 4th decade with an equal male to female ratio was the commonest pattern of arthritis among Chinese, Indians, and Malays. Clinically apparent lumbar spondylitis was significantly more common in Indians than Chinese (10/11 vs 11/20, respectively; p = 0.046), although the prevalence of lumbar spondylitis was similar in all ethnic groups. Eighty-nine percent of subjects required nonsteroidal antiinflammatory drugs and 51% required disease modifying antirheumatic drugs at some time for control of joint disease. PsA was significantly more common among Indians compared to the ethnic distribution of the Singapore population (p < 0.000001). Multiple logistic regression identified Indian ethnicity as a risk factor for the development of PsA (OR 2.39, 95% confidence interval 1.02 to 5.60). CONCLUSION: The commonest pattern of PsA in all ethnic groups was asymmetric polyarthritis. Ethnicity affected the development and presentation of PsA in our series: Indians with psoriasis had double the risk of developing PsA compared to Chinese with psoriasis, and lumbar spondylitis when present in Chinese subjects was asymptomatic in 45%, being detectable only on radiological examination. PMID- 9330939 TI - Ultrasound evaluation of the acromioclavicular joint. AB - OBJECTIVE: To evaluate the value of ultrasonography in assessing arthritic acromioclavicular (AC) joints. METHODS: One hundred twenty-six AC joints of 63 healthy subjects (2 groups) were prospectively examined by ultrasound to determine the-normal limits of capsular distention and the width of the joint space. Thirty-three AC joints of 32 patients with chronic arthritis were evaluated by ultrasound and, for comparison, by radiography, computed tomography (CT), and magnetic resonance imaging (MRI). RESULTS: The mean ultrasonographic distance of the joint capsule from the bone rim was 2.2 mm +/- standard deviation (SD) 0.5 mm in 21-32-year-old control subjects and 2.9 +/- 0.7 mm in 37-81-year old control subjects. The mean width of the joint space was 4.1 +/- 0.9 mm and 3.5 +/- 0.9 mm in the same control groups, respectively. In detecting soft tissue changes in arthritic AC joints MRI was better than ultrasound. In revealing bony surface changes, CT was the best method and radiography was least sensitive but quite specific. Our most prominent finding was that ultrasound is able to exclude joint inflammation; when the ultrasonographic distance of the joint capsule from the bone rim was < 3 mm, there was no synovial hypertrophy or effusion on MRI scans. CONCLUSION: Ultrasound can detect AC joint changes reliably. It is able to exclude joint inflammation. Effusion in the AC joint may reflect inflammation, but may also be a sign of degeneration. PMID- 9330938 TI - Evaluation of a French version of the Bath Ankylosing Spondylitis Disease Activity Index in patients with spondyloarthropathy. AB - OBJECTIVE: To develop a French version of the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and to determine its metric properties in patients with all forms of spondyloarthropathies (SpA). METHODS: A French version of BASDAI was obtained after a translation and back-translation process. Patients fulfilling the European Spondylarthropathy Study Group or Amor criteria for SpA were included. BASDAI of Day 0, Day 1, and, when treatment was changed, Day 8, and other clinical and biological disease activity variables were recorded, along with assessment of disease activity by the physician or the patient. Scalability, reproducibility, sensitivity to change, internal consistency and redundancy, and construct validity of the index were assessed. RESULTS: We studied 293 patients. Good scalability, reproducibility, construct, and internal validity were observed for BASDAI. Sensitivity to change could not be assessed. CONCLUSION: The French version of BASDAI exhibited good metric properties in patients with all forms of SpA, confirming its utility in further clinical research in SpA. However, sensitivity to changes due to drug therapy remains to be assessed. PMID- 9330940 TI - Taking baths: the efficacy of balneotherapy in patients with arthritis. A systematic review. AB - OBJECTIVE: To review English, French, German, and Dutch language studies of the effectiveness of balneotherapy. Balneotherapy (hydrotherapy or spa therapy) is one of the oldest forms of therapy for patients with arthritis. One of the aims of balneotherapy is to relieve pain. METHODS: We performed a systematic review that included randomized and nonrandomized studies. Quality scores of the studies were determined using a criteria list. RESULTS: Most studies report positive findings, but all studies showed methodological flaws. A quality of life measurement was never reported as an outcome measure. None of the randomized clinical trials included intention-to-treat analysis or comparison of effects between groups. CONCLUSION: Because of the methodological flaws a conclusion about the efficacy of balneotherapy cannot be provided from studies we reviewed. We conclude that most flaws found could be avoidable in future research. PMID- 9330941 TI - The limited value of the Health Assessment Questionnaire as an outcome measure in short term exercise trials. AB - OBJECTIVE: To examine the value of the Health Assessment Questionnaire (HAQ) as an outcome measure in short term exercise trials. We studied the association of the objectives of exercise, namely joint mobility, muscle strength, and physical condition, with the HAQ. METHODS: Data for 100 patients with rheumatoid arthritis (RA) included in a study to examine the effect of exercise therapy were used for secondary analysis. Disease activity was determined by the disease activity score (DAS3), pain was scored on a visual analog scale (VAS), and depression was measured by the Dutch Arthritis Impact Measurement Scale. Aerobic capacity (VO2max) estimated from a submaximal ergometer test, grip strength (kPa), isokinetic muscle strength of the knee (Nm/kg), and the Escola Paulista de Medicina range of motion (EPM-ROM) score, a measure of general flexibility, were used as indicators for physical impairments. All variables were entered in a forward multiple regression analysis with the HAQ as dependent variable. RESULTS: The HAQ was significantly correlated with the DAS3 score (r = 0.53), pain (r = 0.51), depression (r = 0.40), joint mobility (r = 0.27), quadriceps strength (r = -0.35), and grip strength (r = -0.50), but not with physical condition. The DAS3 score was first entered in the multiple regression analysis model, followed by pain, quadriceps strength, and grip strength (R2 = 0.45). After 12 weeks of exercise therapy changes in the HAQ were significantly correlated with changes in pain (r = 0.41), in depression (r = 0.33), and in quadriceps strength (r = 0.25), but not with changes in joint mobility or physical condition. CONCLUSION: Physical impairments are weakly associated with the HAQ. The HAQ is not an appropriate instrument to detect changes in physical impairments due to short term exercise therapy. PMID- 9330942 TI - Effects of sonication on articular cartilage in experimental osteoarthritis. AB - OBJECTIVE: To investigate the histological effect of therapeutic ultrasound on arthritic cartilage of rats with various severities of induced osteoarthritis. METHODS: Twenty-seven rats with 3 different stages (Grade I, II, III) of papain induced knee arthritis received 7 min pulse sonication treatment, 3 times/week for 4 weeks. Another 27 rats with the same severity of induced arthritis were studied as controls. The severity of arthritis and related histopathological changes of articular cartilage were evaluated by bone scan and histological findings with hematoxylin and eosin stain. RESULTS: "Severity indexes" based on bone scan decreased after sonication treatment in each study group. Histopathological findings indicated marked cartilage repair in the early stage of induced arthritis (Grade I). However, progressive cartilage damage present in untreated Grade II, III induced arthritis was significantly reduced after sonication. CONCLUSION: Therapeutic ultrasound enhances cartilage repair in the early stage, and has the effect of arresting further deteriorative damage in the later stage of induced arthritis. PMID- 9330943 TI - Osteoarthritic synovial fibroblasts possess an increased level of tumor necrosis factor-receptor 55 (TNF-R55) that mediates biological activation by TNF-alpha. AB - OBJECTIVE: To investigate the presence, number, and level of expression of tumor necrosis factor receptors (TNF-R) in normal and osteoarthritic (OA) human synovial fibroblasts; to examine which receptor isotype mediates the biological response of these cells to TNF-alpha; and to study homologous regulatory mechanisms of TNF-R by TNF-alpha. METHODS: We used radioligand binding assay with [125I]TNF-alpha and flow cytometric analysis with specific antireceptor antibodies to characterize receptor populations, densities, and ligand induced internalization of TNF-R. Inducible cyclooxygenase (COX-2) synthesis, prostaglandin E2 (PGE2) release, and TNF-R shedding (soluble receptors, TNF-sR) were measured after incubation with TNF-alpha the presence or absence of receptor specific blocking antibodies. RESULTS: Although radioligand binding assays showed no difference in the density or affinity of TNF-R in OA synovial fibroblasts compared with normal cells, flow cytometric analysis revealed that OA cells express a significantly higher level of TNF-R55 (p < 0.04) than normal cells. The TNF-R55 was found to be the major receptor species responsible for ligand binding activity, such as COX-2 induction and PGE2 synthesis, since a specific antireceptor TNF-R55 blocking antibody inhibited about 76% of TNF-alpha binding and TNF-alpha stimulated COX-2 induction and PGE2 production. Further experiments revealed that TNF-R55 was the only receptor type internalized after binding TFN alpha, whereas TNF-R75 was concomitantly shed. Moreover, reducing the shedding of TNF-sR, particularly the TNF-sR75, with a synthetic inhibitor decreased TNF-alpha induced PGE2 production. CONCLUSION: TNF-R55 is the major receptor isoform transducing PGE2 and COX-2 responses to TNF-alpha in OA synovial fibroblasts; soluble receptors could be involved in facilitating the binding of TNF-alpha to its receptor. PMID- 9330944 TI - The use of standardized patients in the performance of a needs assessment and development of a CME intervention in rheumatology for primary care physicians. AB - OBJECTIVE: To determine the feasibility of using standardized patients in the performance of a needs assessment and subsequent development of a continuing medical education (CME) intervention in rheumatology for family physicians. METHODS: Eight family physicians and 7 rheumatologists undertook 36 encounters with standardized patients simulating 3 common rheumatologic complaints. Encounters were assessed according to a predetermined score addressing issues of communication skills, clinical diagnosis, investigations, and management. Data obtained were used to develop a small group problem based learning event. RESULTS: Mean scores differed considerably between rheumatologists and family physicians (7.2 vs 3.6; possible range -3 to +9), between individuals within the 2 groups (rheumatologists 2.5 to +9; family physicians -1 to 6.5), and between cases. Specific areas of individual and group need for the development of a subsequent CME intervention were identified and included communication skills, clinical skills in the development of a differential diagnosis, appropriate use and interpretation of investigations, and appropriate management. CONCLUSION: Standardized patients can be effectively used to undertake a needs assessment in rheumatology. Specific data can be used to develop a small group problem based learning program with objectives meeting the identified needs and using the same standardized patients. PMID- 9330945 TI - A comparison of behavioral and educational interventions for fibromyalgia. AB - OBJECTIVE: To compare a comprehensive behavioral intervention with an education/control condition in the treatment of patients with fibromyalgia (FM), and to explore the role of mediators of clinical improvement in both groups. METHODS: The effects of the behavioral and education/control interventions were evaluated across a 10 week treatment period and at 6 month followup on measures of pain, depression, disability, pain behaviors, and intervening variables. The behavioral intervention focused on the development of diverse pain coping skills, while the education/control condition presented information on a range of health related topic without emphasizing skill acquisition. RESULTS: Although improvement across time was found in depression, self-reported pain behaviors, observed pain behaviors, and myalgia scores, no differences in these criteria were found between the behavioral and education/control conditions. Multiple regression analyses revealed that changes in helplessness and passive coping were associated with improvement in a number of clinical outcomes. CONCLUSION: The findings illustrate the value of psychoeducational interventions in decreasing the psychological and behavioral effect of FM, and the value of reducing dysfunctional coping and helplessness in future intervention research. PMID- 9330946 TI - Psychosocial factors associated with complementary treatment use in fibromyalgia. AB - OBJECTIVE: To examine the frequency and predictors of reported complementary treatment use in a sample of 111 subjects with fibromyalgia (FM). The perspective was adopted that complementary treatment use represents a form of medical help seeking that may be subject to a variety of biological, social, and psychological influences. METHODS: Patients with FM were recruited from community and university based clinics and support groups throughout the greater San Diego, California, area. Patients participated in a comprehensive evaluation of their pain, psychological functioning, and disability prior to their potential involvement in a clinical trial designed to help them copy with their condition. They were also administered a rheumatological evaluation to verify their FM and a 20 item questionnaire to assess their use of complementary treatment strategies specifically for coping with FM. RESULTS: Ninety-eight percent of the sample reported the use of at least one strategy over the preceding 6 months. Exercise, bed rest, vitamins, heat treatment, and spirituality/praying were the most frequently used strategies by subjects on a daily basis. Multiple regression analysis revealed that lower age, higher pain, and higher disability were uniquely associated with higher complementary treatment use. The Pain Rating Index, a measure of the subjective severity of pain from the McGill Pain Questionnaire, proved highly significant in explaining the relationship between pain and questionnaire scores. Pain coping strategies and quality of social support did not predict complementary treatment use. CONCLUSION: The findings suggest that poor clinical status is a major predictor of complementary treatment use in FM. However, longitudinal research is recommended to clarify the relationship between clinical status and help-seeking patterns in patients with FM over time. PMID- 9330947 TI - Sleep, daytime symptoms, and cognitive performance in patients with fibromyalgia. AB - OBJECTIVE: To assess sleep, daytime symptoms, and cognitive performance in patients with fibromyalgia (FM). METHODS: Ten female patients with FM (mean age 32 yrs) and a matched, noncomplaintive comparison group (n = 9; mean age 30 yrs) spent 2 nights in the sleep laboratory. After the 2nd night, subjects completed a computerized 20 min battery of self-assessment and performance tests at hourly intervals from 07:00 to 20:00 h. RESULTS: Patients with FM spent more time in stage 1 sleep; however, there were no group differences on any other sleep measures. They reported greater sleepiness, more fatigue, more pain, more negative mood, and lower accuracy on performance tasks across a 14 h day. The FM group was slower in speed, but not impaired in accuracy, on performance of complex tasks, i.e., grammatical reasoning, serial addition/subtraction, and a simulated multi-task office procedure. CONCLUSION: Patients with FM have diurnal impairment in speed of performance on complex cognitive tasks, which accompany light stage 1 electroencephalographic (EEG) sleep and their experience of diffuse pain and nonrestorative sleep symptoms of sleepiness, fatigue, and negative mood. PMID- 9330948 TI - The earnings, income, and assets of persons aged 51-61 with and without musculoskeletal conditions. AB - OBJECTIVE: To describe the personal and family earnings, income, and assets of persons with musculoskeletal conditions. METHODS: This study uses the Health and Retirement Survey, a national, community based probability sample of persons 51 61 years of age and their spouses in 1992 to estimate earnings, income, and assets (by kind) in the years immediately prior to the normal age of retirement. RESULTS: Fifty-nine percent of persons 51-61 years of age (13.76 million) report one or more musculoskeletal condition; of these 38% (8.74 million) also report at least one comorbid condition and 21% (5.02 million) report no such comorbidity. Persons with musculoskeletal conditions and comorbidity report 18% lower family earnings, 15% lower family income, and 35% fewer assets than the average among all persons these ages. Persons with musculoskeletal conditions and no comorbidity have earnings, incomes, and assets closer to the average among their peers. CONCLUSION: Persons with musculoskeletal conditions and comorbidity have lower earnings and incomes now and fewer assets with which to face the future than the remainder of their peers. PMID- 9330949 TI - Prevalence and outcome of uveitis in a regional cohort of patients with juvenile rheumatoid arthritis. AB - OBJECTIVE: To determine the prevalence and outcome of chronic uveitis in patients with juvenile rheumatoid arthritis (JRA). METHODS: A retrospective analysis of 760 patients with JRA followed in 4 pediatric rheumatology centers. Patients with chronic uveitis were identified and their medical and ophthalmologic records were reviewed. RESULTS: Seventy-four patients with uveitis were identified. The prevalence of uveitis was 9.3%. The mean interval from the onset of JRA to the onset of uveitis was 21 months, and 90% of the patients who developed uveitis did so within the first 4 years of their disease. Visual complications (synechiae, band keratopathy, cataract, or glaucoma) developed in 31% of the patients with uveitis. Complications were more common in patients who presented with uveitis early in the course of their JRA. Complications were also more common in antinuclear antibody (ANA) negative than in ANA positive patients. Visual loss to 20/50 or worse occurred in only 11% of patients with uveitis, and no patient became blind. CONCLUSION: In a very large cohort of patients with JRA, uveitis was uncommon and poor visual outcome was rare. Visual complications did not necessarily result in a poor outcome. PMID- 9330951 TI - Liver transplant in adult Still's disease. AB - Adult Still's disease (ASD) is an uncommon form of polyarthritis associated with numerous systemic manifestations, including hepatic involvement. Rarely, liver involvement can be fatal. We describe the case of a young man with ASD with terminal liver failure who required a life saving liver transplant. PMID- 9330950 TI - Hodgkin's lymphoma in systemic onset juvenile rheumatoid arthritis after treatment with low dose methotrexate. AB - We describe the occurrence of malignant lymphoma as a possible complication of immunosuppression associated with low dose methotrexate (MTX) therapy for juvenile rheumatoid arthritis (JRA). A 6-year-old girl with systemic onset JRA who had received low dose MTX therapy for 16 months developed diffuse peripheral lymphadenopathy and enlargement of the lymph nodes in the mediastinum, hilum of the lungs, and liver. Lymph node histology disclosed mixed cellularity Hodgkin's lymphoma; the neoplastic cells were positive for CD30 and CD15, but negative for Epstein-Barr virus RNA or EBV latent membrane protein. After chemotherapy, the girl had complete remission of her disease lasting for 18 months; however, the disease relapsed and autologous peripheral stem cell transplantation was performed. Although the occurrence of lymphoma may be associated with autoimmune diseases, our observations suggest that in pediatric patients, the increasing use of low dose MTX therapy for JRA may be an additional factor for the development of lymphoproliferative disease. PMID- 9330952 TI - Polyarteritis nodosa of the pericardium: antemortem diagnosis in a pericardiectomy specimen. AB - A 74-year-old white woman with diffuse myalgias, low grade fever, and pericardial effusion was found to have polyarteritis nodosa (PAN) in a pericardiectomy specimen. This diagnosis was confirmed at autopsy. Although pericardial involvement in PAN has been described, this is the first report of PAN diagnosed in a pericardiectomy specimen. PMID- 9330953 TI - Colchicine induced rhabdomyolysis. AB - A 59-year-old man with chronic obstructive pulmonary disease (COPD), atrial fibrillation, and gout developed acute dyspnea, cough, and diffuse muscle aches and pains. He had commenced colchicine (0.6 mg b.i.d. p.o.), for the first time, one month earlier for recurrent gout attacks. Clinical examination revealed atrial fibrillation, an exacerbation of his pulmonary disease, tender muscles, especially calves, and diffuse muscle weakness. Laboratory results included creatinine phosphokinase 6961 U/l (1% MB), microscopic hematuria, myoglobinuria, elevated creatinine 1.6 mg/dl, and blood urea nitrogen 17 mg/dl. COPD and atrial fibrillation were treated and colchicine was discontinued. The patient made a full recovery. This 2nd reported case of colchicine induced rhabdomyolysis is the first reported in the treatment of gout. PMID- 9330954 TI - Different course of reactive arthritis in two HLA-B27 positive brothers with fatal outcome in one. AB - During an outbreak of Yersinia pseudotuberculosis III, one of two HLA-B27 positive brothers developed reactive arthritis (ReA), mild at first, but later severely destructive and ultimately fatal. The reactivation of ReA was possibly triggered by an oral polio vaccine. The cause of death was severe secondary amyloidosis. The other brother was exposed to the same Y. pseudotuberculosis strain but did not develop any disease during or after the outbreak. However, he later developed ReA due to a Salmonella infection, with a benign course. PMID- 9330955 TI - Methotrexate osteopathy. PMID- 9330956 TI - Chloroquine and cytokines. PMID- 9330957 TI - Hypoglycemic syncope attack in a patient with rheumatoid arthritis. PMID- 9330959 TI - Lytic vertebral lesions in a patient with AIDS: a new case of Kaposi's sarcoma involving bones. PMID- 9330958 TI - Nephrotic syndrome and renal failure induced by tiopronin in patients with rheumatoid arthritis. PMID- 9330960 TI - Interobserver variation in quantitative analysis of hand radiographs in rheumatoid arthritis: comparison of 3 different reading procedures. PMID- 9330961 TI - Frequency of stages of Alzheimer-related lesions in different age categories. AB - Alzheimer's disease is a relentlessly progressing dementing disorder. Major pathological hallmarks include extracellular deposits of amyloid protein and intraneuronal neurofibrillary changes. No remissions occur in the course of the disease. Initial amyloid deposits develop in poorly myelinated areas of the basal neocortex. From there, they spread into adjoining areas and the hippocampus. Deposits eventually infiltrate all cortical areas, including densely myelinated primary fields of the neocortex (stages A-C). Intraneuronal lesions develop initially in the transentorhinal region, then spread in a predictable manner across other areas (stages I-VI). At stages I-II, neurofibrillary changes develop preferentially in the absence of amyloid deposits. A proportion of cases shows early development of amyloid deposits and/or intraneuronal changes. Advanced age is thus not a prerequisite for the evolution of the lesions. Alzheimer's disease is an age-related, not an age-dependent disease. The degree of brain destruction at stages III-IV frequently leads to the appearance of initial clinical symptoms. The stages V-VI representing fully developed Alzheimer's disease are increasingly prevalent with increasing age. The arithmetic means of the stages of both the amyloid-depositing and the neurofibrillary pathology increase with age. Age is a risk factor for Alzheimer's disease. PMID- 9330962 TI - Plaques and tangles: searching for primary events in a forest of data. PMID- 9330963 TI - Neuropathological staging of Alzheimer-related lesions: the challenge of establishing relations to age. PMID- 9330964 TI - Frequency of stages of Alzheimer-related lesions in different age categories: concurrences and cautions. PMID- 9330965 TI - Frequency of stages of Alzheimer-related lesions in different age categories. PMID- 9330966 TI - The biological significance of neuropathological lesions in Alzheimer's disease. PMID- 9330967 TI - The value of cross sectional neuroanatomical studies as a conceptual framework for prospective clinicopathological studies. PMID- 9330968 TI - The natural history of Alzheimer neurofibrillary tangles and amyloid deposits. PMID- 9330969 TI - The challenge of characterizing normal brain aging in relation to Alzheimer's disease. PMID- 9330970 TI - Unbiased estimation of neuronal numbers in the human nucleus coeruleus during aging. AB - The total number of the neuromelanin-containing neurons of the nucleus coeruleus was determined by means of a newly developed unbiased stereological counting scheme and a low-cost apparative set-up. The individuals (n = 20, age from 49-98 years) included in this study were carefully examined for absence of neurological or psychiatric disorders. However, minor Alzheimer's disease-related neurofibrillary changes occurred in some of the individuals of higher age and these changes were staged. The mean number of neurons per side of the nucleus coeruleus was 15,731 +/- 3,408 SD with a range from 11,737 to 25,319. In three individuals, we compared the left and right nuclei and did not observe significant side differences between the numbers of neurons. There was no correlation between the age of the individuals and the cell number. Also, no correlation was detected between the cell number and the staged occurrence of minor neurofibrillary changes of the Alzheimer type. Due to the novel counting method, the determination of the total cell number took less than 2 h per case. PMID- 9330971 TI - Spatial, temporal and numeric analysis of Alzheimer changes in the nucleus coeruleus. AB - The distribution of neurofibrillary tangles in the nucleus coeruleus was topographically and quantitatively analyzed. The topographical analysis showed statistically significant differences with regard to the distribution of neurofibrillary tangles in the dorsal-ventral and medial-lateral axes. More neurofibrillary tangles were found to be located in the dorsal and medial regions than in ventral and lateral areas. No significant difference in neurofibrillary tangle content was found between the rostral and the caudal areas of the nucleus coeruleus. Neurofibrillary tangle formation begins in the central parts of the nucleus coeruleus. The total number of neuromelanized neurons in the nucleus coeruleus was determined using a modern, unbiased sampling scheme and related to the cortical stage of Alzheimer's disease-related neurofibrillary changes present. A statistically significant reduction (50%) in nucleus coeruleus neurons was evident only in cases meeting the histopathological criteria for Alzheimer's disease. The extent of reduction in the total number of neurons in the nucleus coeruleus did not correlate with the number of neurofibrillary tangles observed. Our data suggest that despite the relatively early susceptibility of the nucleus coeruleus to neurofibrillary tangle formation, significant neuronal loss appears to occur much later, with an estimated average delay time of at least 25 years. Nonetheless, comparison of the topographical pattern of neurofibrillary tangle formation and cell loss indicates that neuronal loss is tangle-related. PMID- 9330972 TI - Vesicular acetylcholine transporter density and Alzheimer's disease. AB - We have evaluated the vesamicol analogue meta-[125I]iodobenzyltrozamicol {(+) [125I]MIBT} as a probe to assess cholinergic terminal integrity in the human temporal cortex. Saturation binding analysis, using 5-aminobenzovesamicol (ABV) to define nonspecific binding, revealed a high-affinity binding site with a Kd value of 4.3 +/- 1.2 nM in the temporal cortex of the young control subjects. Similar affinity values were observed for (+)-[125I]MIBT binding in aged control subjects (Kd = 3.4 +/- 0.5 nM) and AD patients (Kd = 3.0 +/- 0.8 nM). In contrast, Bmax values for young subjects, aged controls and AD patients were 31.2 +/- 6.3, 17.0 +/- 2.0 and 9.4 +/- 1.6 pmol/g, respectively, clearly reflecting significant reductions in (+)-[125I]MIBT binding site density with aging and age related neuropathology. Moreover, the decrease in (+)-[125I]MIBT binding was correlated with choline acetyltransferase activities (r = 0.72) in the AD temporal cortex. These results suggest that when selective ligands are used, the vesicular acetylcholine transporter can be a useful marker protein for assessing the loss of cholinergic projections in AD and related disorders. PMID- 9330973 TI - Molecular and cellular characterization of the membrane attack complex, C5b-9, in Alzheimer's disease. AB - The membrane attack complex, C5b-9, is of considerable importance in many inflammatory reactions. It is the terminal, cytolytic component of both classical and alternative pathway activation, and its presence presupposes other potentially destructive complement constituents, including anaphylotoxins and opsonins. We have characterized C5b-9 and its C9 constituent in the Alzheimer's disease (AD) and nondemented elderly (ND) brain using immunohistochemistry at the light and electron microscopic levels, Western blot analysis, and the reverse transcriptase polymerase chain reaction. We have also conducted in vitro ELISA assays of amyloid beta-peptide-stimulated SC5b-9 production. C5b-9 is abundantly present in Alzheimer's disease cortex, associated with neurofibrillary tangle containing neurons, dystrophic neurites within neuritic plaques, and neuropil threads, but is weakly detected, if at all, in nondemented elderly cortex under the same conditions. Staining of Alzheimer's disease sections is abolished both by deletion of primary antibody or preabsorption with purified SC5b-9. PMID- 9330974 TI - Cortisol enhances non-REM sleep and growth hormone secretion in elderly subjects. AB - Aging is accompanied by a continuous decline in slow wave sleep (SWS) and in growth hormone (GH) secretion, particularly during the sleeping period. Because short-term pulsatile administration of cortisol increases GH release and SWS in young adults, we wondered whether similar effects can be induced also in elderly men. Hourly injections of cortisol between 1700 and 600 h increased stage 2 and SWS and decreased rapid eye movement sleep. Spectral analysis revealed significant increases in delta and theta power. Cortisol infusions increased the GH secretion prior to sleep onset, but remained largely unchanged during sleep. Thus, sleep EEG and GH release are modulated by cortisol administration in a manner similar to that in young subjects, but to a lesser extent. The stimulatory effect of cortisol on both GH release and SWS points to a mechanism involving glucocorticoid-enhanced production and release of GH-releasing hormone that activates pituitary GH release and simultaneously antagonizes the effects of corticotropin-releasing hormone and somatostatin. PMID- 9330975 TI - Age-related increases in brain monoamine oxidase B in living healthy human subjects. AB - Several studies of human brain postmortem report that monoamine oxidase B (MAO B) increases with age and it has been proposed that this increase reflects age associated increases in glial cells. We measured brain MAO B in a group of normal healthy human subjects (n = 21; age range 23-86; 9 females and 12 males; nonsmokers) using [11C]L-deprenyl-D2 and positron emission tomography. Brain glucose metabolism was also measured with 18FDG in 15 of the subjects. MAO B increased (p < 0.004) in all brain regions examined except the cingulate gyrus. In contrast, subjects showed the expected regional age-related decreases in blood flow and metabolism. In the 15 subjects in whom both MAO B and LCMRglu was measured, there was a trend (p < 0.03) toward an inverse association between brain glucose metabolism and MAO B activity in the frontal and parietal cortices. Although the age-related increase in brain MAO B in living subjects is consistent with postmortem reports, the degree of increase is generally lower. PMID- 9330976 TI - Cerebral glucose metabolism and memory in aged rhesus macaques. AB - Positron emission tomography and the glucose metabolic tracer [18F]fluorodeoxyglucose were used to evaluate the relationship between regional cerebral metabolic rates for glucose (rCMRglc), age, and performance on a delayed response (DR) test of memory in the aged monkey. Eleven aged animals, 21-26-years old, were included in the analysis. Regional CMRglc, normalized to values for the entire brain, were determined for the dorsal prefrontal cortex, orbitofrontal cortex, hippocampus, and temporal cortex. The aged animals exhibited significant DR deficits relative to a cohort of normal young monkeys. Variability in DR performance among the aged subjects was significantly correlated with relative hippocampal rCMRglc, and chronological age was a reliable predictor of orbitofrontal rCMRglc ratios. This pattern of results suggests that DR impairments in the aged monkey may partly reflect age-related dysfunction distributed among multiple limbic system structures that participate in normal learning and memory. Overall, the findings support the use of positron emission tomography in efforts to define the relationship between cognitive performance, age, and brain physiology in nonhuman primates. PMID- 9330977 TI - Age-related decrease in the N-methyl-D-aspartateR-mediated excitatory postsynaptic potential in hippocampal region CA1. AB - Glutamatergic fast synaptic transmission is known to be altered with age in a region-specific manner in hippocampus of memory-impaired old rats. In the present experiment, presynaptic fiber potentials and non-N-methyl-D-aspartate (NMDAR) and NMDAR-mediated synaptic responses in CA1 were compared in three ages of behaviorally characterized male F-344 rats. In the CA1 region, old rats showed approximately equivalent reductions in non-NMDAR- and NMDAR-excitatory postsynaptic potential amplitudes for a given size of presynaptic fiber potential. There was no change in magnitude of the presynaptic response itself at any stimulus level. These results are consistent with the hypothesis that there is a reduction in the number of Schaffer collateral synapses per presynaptic axon. This pattern of results in CA1 is very different from what is known to occur at the perforant path-granule cell synapse. In fascia dentata the non-NMDAR mediated excitatory postsynaptic potential is increased in amplitude, although the NMDAR-mediated excitatory postsynaptic potential is reduced for a given presynaptic input. These data suggest that age-related functional alterations in neurotransmitter receptor subtypes occur differentially between closely-related anatomical subregions. PMID- 9330978 TI - Consensus recommendations for the postmortem diagnosis of Alzheimer's disease. The National Institute on Aging, and Reagan Institute Working Group on Diagnostic Criteria for the Neuropathological Assessment of Alzheimer's Disease. AB - This report summarizes the consensus recommendations of a panel of neuropathologists from the United States and Europe to improve the postmortem diagnostic criteria for Alzheimer's disease. The recommendations followed from a two-day workshop sponsored by the National Institute on Aging (NIA) and the Ronald and Nancy Reagan Institute of the Alzheimer's Association to reassess the original NIA criteria for the postmortem diagnosis of Alzheimer's disease published in 1985. The consensus recommendations for improving the neuropathological criteria for the postmortem diagnosis of Alzheimer's disease are reported here, and the "position papers" by members of the Working Group that accompany this report elaborate on the research findings and concepts upon which these recommendations were based. Further, commentaries by other experts in the field also are included here to provide additional perspectives on these recommendations. Finally, it is anticipated that future meetings of the Working Group will reassess these recommendations and the implementation of postmortem diagnostic criteria for Alzheimer's disease. PMID- 9330979 TI - Neuropathological criteria for the diagnosis of Alzheimer's disease: are we really ready yet? AB - The specific diagnosis of AD as a particular dementia from which a patient suffered assumes, debatably, a reasonably pure clinicopathological entity in which the same concatenation of lesions will not be encountered in others dying with a similar clinical disorder. Statistically complex computations such as multivariate analyses of morphometric data from our laboratory and similar attempts in Swedish and British series may not prove pragmatic for pathological confirmation. The Braaks' schema posits six stages in the evolution of AD. Unfortunately, application of this model to 50 British autopsies cannot reliably identify those cases clinically diagnosed as demented. Furthermore, lack of universal definition for each of the probable lesional subtypes augments the difficulty devising a quantitative consensus. Disease stage refers to a progressive increase in anatomical (geographic) extent of involvement, whereas, grade refers to a progressive increase in severity of affliction within any one site. There is only a tendency for stage and grade to progress in parallel. Nor is it obligatory that either always does progress. More energies should be concentrated upon determining which histopathological abnormality is most injurious to neuronal integrity. Dutch workers opine that in both normal aging and AD, claims of massive, neocortical nerve cell loss may have been based on inadequate morphometry and/or a loss of markers. Requiring urgent resolution is whether cellular changes seen in brains of aging normals represent merely the earliest phase of typical AD (and therefore a good model for Alzheimer pathogenesis), or rather reflect a totally different aging syndrome distinct from AD. We have proposed that abnormalities in the hippocampal formation (with or without neocortical neuronal lesions) may underlie a decline of all higher cognitive functions in senile dementia Alzheimer type. West and colleagues optical disector approach likewise shows that neurodegeneration associated within aging individuals' hippocampi is quantitatively and qualitatively distinct from the neuronal loss in AD. Clinical confreres' imprecision whether or when to term subtle cognitive loss "incipient AD" is understandably mirrored by residual neuropathological struggles to dichotomize such brains as "normative aging" distinct from "putative AD." PMID- 9330980 TI - Neuropathological criteria for the diagnosis of Alzheimer's disease. AB - The definitive diagnosis of Alzheimer's disease (AD) is made at autopsy by the presence of abundant neuritic plaques (NP) and neurofibrillary tangles (NFT) in the neocortex, entorhinal cortex, and hippocampus. The two criteria most frequently used by neuropathologists for the diagnosis of AD over the past 12 years are those described by Khachaturian and the Consortium to Establish a Registry for Alzheimer's Disease. Though both have been useful, they have weaknesses and lack validation. The majority of recent studies has shown that NFT in the entorhinal cortex, hippocampus, and neocortex and NP in the neocortex correlate best with severity of dementia in AD. The criteria recommended by the Workshop on Diagnostic Criteria for the Neuropathological Assessment of AD uses semiquantitation of NFT and NP in the neocortex, adds evaluation of the hippocampus and entorhinal cortex, places emphasis on coexisting lesions such as vascular lesions and Lewy bodies, and establishes criteria for general pathologists and more rigorous criteria for the AD research setting. These criteria will require further refinement and validation. PMID- 9330981 TI - Neuropathological diagnosis of Alzheimer's disease: a perspective from longitudinal clinicopathological studies. AB - Alzheimer's disease (AD) is the most common cause of primary progressive dementia. It is defined by its pathological features, because the clinical syndrome of dementia lacks specificity. Although the brain in AD has many structural alterations, the two cardinal pathological features and sine qua non of AD are senile plaques and neurofibrillary degeneration. The latter takes the form of neurofibrillary tangles composed of paired helical filaments, as well as degenerating neurites within the neuropil ("neuropil threads") and within a diagnostically significant subset of SP, referred to as "neuritic plaques." All SP contain amyloid, but not all have pair helical filament-type neurites. The presence of even a small number of plaques with paired helical filament-type neurites in the neocortex is associated with cognitive impairment, and this lesion may be the most specific histopathological feature of AD. Although these observations suggest that the major difference between SP in aging and AD relate to differences in neuritic degeneration, more recent studies have also indicated that amyloid deposits are biochemically heterogenous and that amyloid deposits in aging may be different from those in AD. As more specific markers become available for recognizing AD-specific structural lesions, refined neuropathological diagnostic criteria will evolve. In the meantime, a practical neuropathological approach to the diagnosis of AD requires both widespread neocortical SP and an advanced stage of neurofibrillary degeneration. Using these criteria, it is very unlikely that AD will be diagnosed in an individual who was not demented in life. The rationale for adopting this conservative approach is that our knowledge is incomplete with respect to fundamental differences between the lesions in aging and AD. PMID- 9330982 TI - The neuropathological diagnosis of Alzheimer's disease: clinical-pathological studies. AB - The neuropathological diagnosis of Alzheimer's disease currently relies on quantitative or semiquantitative criteria of senile or neuritic plaques that are adjusted for age and for the presence or absence of a clinical history of dementia. Based on clinical-pathological correlation studies, I will argue that neuropathological assessment should stand independently of clinical history and instead should describe brain lesions in the context of the topography and natural history of the disease. Only probabilistic estimates about the presence or absence of dementia can be made from a neuropathological examination, especially in the setting of Alzheimer disease lesions plus other pathological alterations such as Lewy bodies or infarcts. Moreover, I will argue that any neurofibrillary tangles or senile plaques are inherently pathological entities, even if clinically silent and so "incidental" neuropathological findings. Because the intensity and location of neurofibrillary tangles, rather than senile plaques, appears to correlate most closely with clinical symptoms, I suggest using a staging system that highlights this information rather than using absolute numerical cut-offs for diagnostic purposes. PMID- 9330983 TI - Diagnosis and staging of Alzheimer disease. AB - Rather than determining lesions "threshold" between "normal" cases and patients, we prefer to use clinicopathological correlations, assigning a given intellectual deficit to a given amount of lesions with a chosen level of probability. Because large amounts of A beta diffuse deposits may be found in the absence of dementia, we think advisable not to take them into account for the diagnosis. The diffusion of the neurofibrillary tangles in the paralimbic, limbic and isocortical areas (described by braak and Braak stages or by the number of areas containing tangles) and the density of isocortical senile plaques (A beta focal deposits) as assessed by the CERAD protocol are both correlated with the intellectual status but give complementary information. They should thus be jointly used. We analyzed the variability of the lesions counts, their coefficients of error, and their causes, as a first step toward standardization. We have shown, however, that semiquantitative estimates are presently more reproducible than quantitative measures. PMID- 9330984 TI - Diagnostic criteria for the neuropathological assessment of Alzheimer's disease: current status and major issues. AB - A Consensus Conference focusing on Alzheimer's disease (AD) took place in November 1996 to recommend uniform evaluation procedures and diagnostic criteria, co-sponsored by the National Institute on Aging and the Reagan Institute of the Alzheimer's Association. In conjunction with this conference, we reviewed diagnostic practices in current use, together with various neuropathological criteria proposed since 1985. Difficulties were identified in developing "gold standard" criteria for diagnosis and case classification of AD based upon the current state of knowledge. Working criteria for use within research contexts were proposed that acknowledged the realities of scientific limitations by inclusion of a broad and heterogeneous category of "uncertain" cases. (Eventually, methods will be developed for identifying these cases as preclinical AD, dementia due to multiple causes or non-AD, but this is not now possible.) Within applied contexts, the use of CERAD guidelines was supported. Finally, recommendations generated at the Consensus Conference were discussed, emphasizing the rapid pace of recent scientific advancement and the need for ongoing empirical reevaluation and modification of the Group's proposal. PMID- 9330985 TI - A commentary on the diagnostic criteria for the neuropathological assessment of Alzheimer's disease. AB - Apart from providing a potential framework for a universally acceptable protocol for the pathological diagnosis of Alzheimer's disease, the Working Party has dispelled one of the major myths of so-called normal aging, namely that minor degrees of Alzheimer-type change (i.e., plaques and tangles) are not the product of growing older alone but should always be regarded as pathological, as they probably represent the tissue substrate of incipient Alzheimer's disease. Establishment of the pathological criteria for such preclinical disease will be of equal importance as that associated with clinical change. PMID- 9330986 TI - Diagnostic criteria for the neuropathologic assessment of Alzheimer's disease. AB - The provisional criteria proposed in 1985 by Khachaturian et al. emphasized numbers of plaques and neglected tangles, as did CERAD (Consortium to Establish a Registry for Alzheimer's Disease). The decision to set an arbitrary number of plaques as "pathologic" assumed that some neuritic plaques are a normal phenomenon in the aging brain. Neuritic plaques and neurofibrillary tangles are age-related lesions, but they are pathologic (i.e., lesions) no matter how many there are. In a clinically demented patient without vascular or other neurodegenerative lesions, a clinico-pathologic diagnosis of AD (a clinico pathologic entity) can be made with a high level of confidence by demonstrating, and without counting, plaques and tangles. The vast majority of AD cases are straightforward, and diagnostic lesions can be appreciated with a simple silver stain. If patients' histories are unknown or uncertain, the clinical significance of the observed plaques and tangles must remain debatable. This is the essence of the consensus statement with which I wholeheartedly agree. In such cases without a dementia history, one can offer a neuropathologic diagnosis of Senile or Pre senile Cerebral Disease (not "dementia") of the Alzheimer type. Precise clinico pathologic correlations and some quantitative measures are needed for elucidating the pathogenesis of AD and for establishing a primary dementing diagnosis when AD is mixed with other dementing diseases. These correlations must be based on periodic and fairly extensive neuropsychological testing followed shortly thereafter by a detailed postmortem neuropathologic evaluation. PMID- 9330987 TI - Proposals for re-evaluation of current autopsy criteria for the diagnosis of Alzheimer's disease. AB - Defining criteria for the postmortem diagnosis of Alzheimer's disease (AD) has proven difficult due to the phenotypical heterogeneity of the disease, the absence of a specific disease marker and an overlap of AD neuropathology with that observed in a number of nondemented aged individuals. Even though the role of plaques and tangles in the pathogenesis of AD remains undetermined, a host of clinicopathological correlative studies have shown that both lesions, if present in sufficient numbers-particularly in the neocortex-are still to be considered the best morphological signposts for the disease. All currently used criteria for the neuropathologic diagnosis of AD have some weaknesses and need to be reestablished and revalidated. Multivariant analysis in a personal autopsy series of elderly subjects revealed significant correlations between psychostatus and both the CERAD criteria and Braak staging of neuritic Alzheimer-type lesions, and less concordance with the National Institutes of Aging and Tierney criteria. We propose a set of histopathologic diagnostic criteria for both definite and preclinical AD that rely on various constellations of both different types of plaques, except diffuse amyloid deposits, and neurofibrillary tangles, in allocortical and isocortical areas considering their topographic pattern. This set of criteria encompasses phenotypic variations of the pathology and takes into account the chronic, progressive course of AD. It allows the detection of preclinical disease in subjects in whom dementia is not reported and includes those cases in the morphological gray zone between "normal" aging and full fledged AD that practicing neuropathologists consider the most problematic. The set of criteria includes guidelines concerning tissue sampling and processing, and standardized staining methods that should allow neurologists to minimize interrater and interlaboratory variability in the assessment of morphologic lesions and the diagnosis of AD. PMID- 9330988 TI - Diagnostic criteria for Alzheimer's disease. AB - This paper summarizes changes that distinguish early Alzheimer's disease (AD) from nondemented aging based on 49 well characterized cases (30 nondemented, 10 very mildly demented, and 9 severely demented). Tangles were found in all nondemented cases (aged 54 to 88) concentrated in limbic structures. The probability of high tangle density increases with age, even in the absence of plaques or dementia. Based on plaques, nondemented cases can be divided into three groups: 1) cases younger than 73 years of age with one-third of older cases had no plaques; 2) about one-half of cases over 74 years of age had a few diffuse plaques in restricted patches in the neocortex; 3) about one-quarter of cases over 74 years of age had many neuritic and diffuse plaques throughout the neocortex; these may represent "preclinical" AD. Very mildly demented cases had high concentrations of neuritic and diffuse plaques in the neocortex and tangles in limbic structures. The observations indicate that the minimal diagnostic criterion for AD is plaques throughout the neocortex together with neurofibrillary changes (tangles in limbic structures and neuritic plaques in cortex). Tangles are a necessary but not sufficient criterion. PMID- 9330989 TI - Position paper on diagnostic criteria for Alzheimer disease. AB - The lesions of Alzheimer disease (AD) consist of synapse and neuron loss associated with progressive deposition of amyloid as diffuse and neuritic plaques and accumulating tau abnormalities in the form of neurofibrillary tangles and neuropil threads. Diagnostic criteria for Alzheimer disease constitute arbitrary cut-off levels above which AD is deemed to exist, and below which lesser amounts of the same abnormalities are relegated to the nebulous category of aging changes. Demanding neocortical tangles for a diagnosis of AD sacrifices sensitivity on the altar of specificity, since, while such lesions usually represent an advanced stage in the orderly evolution of AD, lighter burdens of plaque-predominant AD pathology with tangles confined to the medial temporal lobe can cause dementia when associated with concomitant synapse loss. Such muted AD pathology typifies the Lewy body variant of AD, and it serves to segregate it from pure Lewy body disease. We endorse the semiquantitative neuritic-plaque based criteria from CERAD for routine diagnosis, and Braak staging with descriptive profiling of AD lesions in a research context. PMID- 9330990 TI - Strategies for improving the postmortem neuropathological diagnosis of Alzheimer's disease. AB - Despite recognition that Alzheimer's disease (AD) is a polygenic and heterogeneous dementing neurodegenerative disorder, there is continued merit in defining the AD phenotype by the presence of progressive cognitive impairments and the pathological brain lesions (senile plaques, neurofibrillary tangles) as originally formulated by Alois Alzheimer. This position paper discusses the rationale for emphasizing the detection of both beta amyloid-rich plaques and tau rich tangles in the next iteration of the neuropathological criteria for the postmortem diagnosis of AD that has been recommended by the Working Group on Consensus Criteria for the Postmortem Diagnosis of AD. Further, it also underlines the need to exploit continuing advances in understanding the pathobiology of plaques and tangles in subsequent iterations of these criteria. It is expected that such efforts, now and in the future, will hasten the development of strategies for the early and accurate antemortem diagnosis of AD as well as the discovery of effective treatments for this common dementing illness of the elderly. PMID- 9330991 TI - Proposal to revise the morphologic criteria for the diagnosis of Alzheimer's disease. AB - Ten years have passed since the drafting of the original National Institute on Aging/American Association of Retired Persons or Khatchaturian criteria for the neuropathologic diagnosis of Alzheimer's disease. In that time, much progress has been made in the study of this disorder. It is clear that although the Khatchaturian criteria have been useful, advances in our understanding of the morphologic substrate of the disease needs to be incorporated in any newly proposed diagnostic criteria. We propose diagnostic criteria to be employed in establishing the diagnosis of Alzheimer's disease in a research setting. Here, we require that a level of density and distribution of senile plaques and neurofibrillary tangles be identified and that other superimposed conditions, such as major cerebral infarcts and/or Parkinson's disease changes, are absent. We also propose separate criteria for diagnosing individuals who die in the early stages of the disease. These latter cases are of extreme research interest and are characterized by a distribution of neurofibrillary tangles that remains primarily restricted to the entorhinal cortex and hippocampus. PMID- 9330992 TI - Diagnostic criteria for neuropathologic assessment of Alzheimer's disease. AB - Prior to any evaluation of morphologic brain changes, a decision must be made whether a given alteration is associated with aging or with disease. Patients with disease-related lesions may be in a clinically silent phase of a disease or show overt symptoms. Neurofibrillary tangles and neuropil threads are the hallmarks of Alzheimer's disease. They should not be considered to be age-related changes, even when they are present only in small numbers. In general, the initial changes consist of neurofibrillary tangles and neuropil threads. Plaques (amyloid deposits and/or neuritic plaques) are consistently present in the end stage of the disease. Initial neurofibrillary tangles and neuropil threads develop at specific cortical predilection sites. The changes then spread in a predictable, nonrandom manner across other portions of the telencephalic cortex. The sequential changes in the distribution pattern of the lesions provide the basis for a staging procedure that takes the slow and gradual progression of the destructive process into consideration. The staging procedure provides accurate diagnoses in the initial stages and even reveals brain changes developing prior to the appearance of clinical symptoms. It is thus advantageous in characterizing nondemented controls. The staging procedure can be carried out easily and does not require knowledge of clinical data, quantitative assessments, or adjustments for the age of the patients. Application of advanced silver techniques (Gallyas, Campbell-Switzer) to demonstrate Alzheimer's disease-related lesions also allows recognition of the hallmarks of other disorders, such as Lewy body disease (Parkinson's disease) and dementia with argyrophilic grains, which frequently co occur with Alzheimer's disease. PMID- 9330993 TI - Commentary on the consensus recommendations for the post mortem diagnosis of Alzheimer's disease. AB - The consensus recommendations for the post mortem diagnosis Alzheimer's disease (AD) highlight the difficulties in establishing a pathological diagnosis in brains from clinically demented individuals with both certainty and uniformity. There is, however, a need for diagnostic guidelines that are relatively simple, inexpensive, and adaptable to general pathologists and different laboratories. The current Consortium to Establish a Registry for Alzheimer's disease (CERAD) criteria and the recommendations in the consensus document giving three probabilistic categories for diagnosis go a long way towards establishing a uniform approach for the diagnosis of AD. However, more uniformity could be adopted in the topography of sectioning to enhance diagnostic and future research comparisons. We also recommend that immunohistochemistry for beta A4 (A beta) amyloid and tau-reactive neurofibrillary changes, in addition to hematoxylin and eosin stains, should become the basis for histological diagnosis. We agree with the guidelines concerning documentation of all AD changes. Until a clearer understanding of the early changes of AD is established, strict observation and recording are the pathologists' best diagnostic skills. The ill-defined diagnostic areas of AD continue to prompt the need for a new method of detection of the underlying pathologic process. PMID- 9330994 TI - The CERAD neuropathology protocol and consensus recommendations for the postmortem diagnosis of Alzheimer's disease: a commentary. AB - CERAD, a multicenter longitudinal study, has developed standardized instruments for the evaluation of individuals clinically diagnosed as having Alzheimer's disease (AD). The CERAD neuropathology protocol not only establishes levels of certainty for AD diagnosis, but also records information on other conditions occurring concomitantly with or misdiagnosed as AD. The protocol has been widely adopted because of its relative simplicity, adaptability, and reliability. Indeed, the Consensus principles proposed are, for the most part, consistent with CERAD guidelines. The recommendation that diagnosis rest upon both neuritic plaque and neurofibrillary tangle frequency/distribution in the neocortex, however, is worrisome. This change will eliminate or downgrade many cases now diagnosed as AD with concomitant Parkinson's disease changes. Reclassifying such cases at this time, without compelling pathobiological justification, is premature. Instead, I recommend retention or modest modification of the current CERAD protocol, and propose that neuropathology data available on autopsies of over 200 CERAD dementia subjects be used for testing potential modifications of the diagnostic algorithm. PMID- 9330996 TI - Research uses of neuropathological data. AB - The study of relationships between neuropathological characteristics and behavioral, structural, chemical, and molecular variables offers immense promise for understanding the basic pathophysiology of Alzheimer's disease. This position paper examines the need for standardized procedures and quantitation if neuropathological data are to be optimally useful among laboratories investigating the biology of Alzheimer's disease. These requirements include standardized fixation, embedding, sectioning, staining, brain regions and sampling methods. In addition, the definition of the structures to be quantified, such as plaque type(s), needs to be rigorously specified. Unbiased stereological methods for quantification should be used. These needs for optimal research utility exceed the needs and practicality for diagnostic purposes, suggesting a two-tiered approach to the neuropathology of Alzheimer's disease: diagnostic and research. PMID- 9330995 TI - Current diagnostic criteria for assessment of neuropathological Alzheimer's disease. AB - Following a critical review of Khachaturian's and other diagnostic criteria of Alzheimer's disease (AD), it is concluded that new, more rigorous criteria are needed. These criteria must be based on objective data, and they should distinguish subjects with mild cognitive impairment from those with normal cognition. Moreover, they should be easy to use. A multicenter study is proposed and outlined to examine the pathology of 3 groups of 20 archival or newly obtained subjects. The three groups should include subjects who are cognitively intact, slightly impaired, and affected by definite AD, as determined by appropriate and timely cognitive tests. PMID- 9330997 TI - Comparison of neuropathologic criteria for the diagnosis of Alzheimer's disease. AB - The National Institute on Aging and Reagan Institute (NIA-RI) criteria, and other neuropathologic criteria for Alzheimer's disease (AD), were compared with the clinical diagnosis of dementia in a well defined population of Catholic sisters. The 47-participant subset examined in this study were college educated and lacked complicating conditions such as brain infarcts or diffuse Lewy body disease. Sixteen participants had a clinical diagnosis of dementia. The NIA-RI criteria imply a perfect correlation between neuritic plaque (NP) density and neurofibrillary tangle distribution. However, NP density often did not coincide with tangle distribution. As a result, it was not possible to categorize many of the participants using the NIA-RI guidelines. The 'high likelihood' category of the NIA-RI criteria for AD research settings (neocortical Braak stage and frequent neocortical NP) had relatively high specificity (90% of nondemented participants did not meet this criteria). However, only half of the demented participants were in this category. Neuropathologic criteria requiring the presence of neocortical tangles (rather than neocortical Braak stage) had relatively high sensitivity, accounting for 87-94% of participants with dementia, but also included 32-35% of nondemented participants. Criteria based on neocortical NP or senile plaques had 100% sensitivity, but a majority of nondemented participants also met these criteria. The results support consideration of both tangles and NP for the neuropathologic diagnosis of AD, but indicate that refinement of the NIA-RI criteria is necessary. A possible refinement is suggested for further consideration. PMID- 9330998 TI - Do we need angiotensin II antagonists to treat hypertensive patients? PMID- 9330999 TI - Candesartan cilexetil: a review of its preclinical pharmacology. AB - Candesartan is a highly potent, long-acting and selective angiotensin II type 1 (AT1) receptor blocker. It is administered orally as the inactive prodrug candesartan cilexetil which is rapidly and completely converted to candesartan during gastrointestinal absorption. In vitro studies have shown that candesartan acts as an insurmountable angiotensin II receptor antagonist, binding tightly to and dissociating slowly from the AT1 receptor. The above characteristics are thought to contribute to the marked and long-lasting antihypertensive effects of candesartan cilexetil in several animal models of hypertension. These included rodent models of renal hypertension in which candesartan cilexetil also demonstrated efficacy equivalent to or greater than enalapril. In other animal models, candesartan cilexetil reduced the incidence of stroke, renal dysfunction and renal disease while reducing cardiac and vascular hypertrophy. Furthermore, candesartan cilexetil conferred some protection against cerebral and renal damage at a dose that had no blood pressure-lowering effect. In toxicity and general pharmacology studies, candesartan cilexetil was shown to possess a 'clean' profile with a large safety margin. Also it did not potentiate chemical- or autocoid-induced cough or anaphylactoid reactions. PMID- 9331000 TI - Pharmacokinetics of candesartan after single and repeated doses of candesartan cilexetil in young and elderly healthy volunteers. AB - Candesartan cilexetil is rapidly and completely hydrolysed to the active compound candesartan during absorption from the gastrointestinal tract. Candesartan is a potent, long-acting, selective angiotensin II AT1 receptor blocker. The pharmacokinetics of candesartan were investigated after single and repeated once daily doses of candesartan cilexetil in the dose range 2-16 mg in both younger (19-40 years) and elderly (65-78 years) healthy volunteers in five studies. Blood pressure, heart rate, and hormones associated with the renin-angiotensin system, and safety of candesartan cilexetil administration were also assessed. Placebo comparisons were made in four studies. Frequent blood samples were collected after the first single dose of candesartan cilexetil, and during the last dosing interval after 1 week repeated once-daily administration. Serum and plasma were analysed for candesartan cilexetil, candesartan and its inactive metabolite, CV 15959, as well as angiotensin I and II, aldosterone, plasma renin activity (PRA) and angiotensin-converting enzyme (ACE) activity. The AUC and Cmax of candesartan showed dose-proportional increases in the dose range of 2-16 mg candesartan cilexetil after both single and repeated once-daily tablet intake, indicating linear pharmacokinetics in both younger and elderly healthy subjects. The pharmacokinetics did not change on repeated dosing and, as expected from the half life of candesartan of approximately 9 h in younger subjects, there was almost no accumulation after repeated once-daily dosing. The time to peak candesartan concentrations after tablet intake was consistently approximately 4 h at all dose levels. Both Cmax and AUC of candesartan were increased after single and repeated once-daily dosing in the elderly compared to younger subjects by approximately 50%. However, no accumulation after repeated once-daily dosing were seen in the elderly. The half-life of candesartan in the elderly (9-12 h) was somewhat longer than in the younger healthy adult volunteers (approximately 9 h) and no gender related differences in the disposition of candesartan were observed. Serum concentrations of CV-15959 were much lower than candesartan, and reached peak serum concentrations later, about 4-9 h after dose intake. The elimination of CV 15959 was somewhat slower than that of candesartan. Candesartan cilexetil, the prodrug to candesartan, was not measurable in serum. No differences in ACE activity or serum aldosterone concentrations were observed between subjects receiving candesartan cilexetil and placebo tablets. Plasma angiotensin I and II concentrations and PRA were augmented after single doses and further increased after 1 week repeated candesartan cilexetil dosing. Single and repeated doses of candesartan cilexetil were well tolerated in the younger and elderly volunteers. Only mild adverse events were recorded, with 'headache' as the most commonly reported event, and no increase in the number of reported adverse events was observed with higher doses of candesartan cilexetil. No clinically significant changes in respect to vital signs, physical examination, ECG, and clinical laboratory tests were observed. PMID- 9331001 TI - Absorption, metabolism and excretion of 14C-candesartan and 14C-candesartan cilexetil in healthy volunteers. PMID- 9331002 TI - Bioavailability of candesartan is unaffected by food in healthy volunteers administered candesartan cilexetil. PMID- 9331003 TI - Pharmacokinetic drug interaction studies with candesartan cilexetil. AB - The aim of this series of studies was to determine the potential for pharmacokinetic interaction between candesartan (administered orally as the prodrug candesartan cilexetil) and hydrochlorothiazide (HCTZ), nifedipine, glibenclamide, warfarin, digoxin or the components of an oral contraceptive formulation. All studies were performed in healthy volunteers using randomised, crossover or add-on study designs. Candesartan cilexetil was administered orally at doses of 8, 12 or 16 mg. The pharmacokinetic parameters were determined for comparator agents and candesartan following administration of each agent alone or in combination. There were no changes in the drug plasma concentrations of nifedipine, glibenclamide, digoxin or oral contraceptives when co-administered with candesartan cilexetil. Co-administration of candesartan cilexetil caused a slight but significant decrease in the AUC of HCTZ. However, the 90% confidence intervals (CI) for AUC ratios for HCTZ when co-administered with candesartan cilexetil were within the defined limits of bioequivalence. Candesartan cilexetil produced a 7% decrease in trough plasma warfarin concentration but this had no effect on prothrombin time. Co-administration of candesartan cilexetil with HCTZ produced a statistically significant increase in the bioavailability and Cmax values for candesartan (18% and 25%, respectively). However, this increase is not considered to be clinically relevant. No other co-administered drug (nifedipine, glibenclamide, digoxin, oral contraceptive) affected the pharmacokinetic parameters of candesartan. Candesartan cilexetil was well tolerated both alone and in combination with the other agents. PMID- 9331004 TI - Pharmacokinetics of candesartan cilexetil in patients with renal or hepatic impairment. AB - Five clinical studies were conducted to investigate the pharmacokinetic profile and safety of candesartan cilexetil in patients with either normal or impaired renal or hepatic function. Participants in these open-label, single- or parallel group prospective studies were administered candesartan cilexetil 8 or 12 mg as a single oral dose and then, in all but one study, as a repeated once-daily oral dose regimen. A total of 94 patients of either gender aged between 18 and 75 years with normal or mild to moderate hepatic dysfunction (Study 1) and normal or mild to moderate/severe renal dysfunction (Studies 2-5) were included. Subjects recruited to all studies evaluating the effect of renal impairment also had some degree of hypertension. Patients with mild to moderate hepatic impairment showed no significant differences in the key plasma pharmacokinetic parameters or plasma protein binding profile of candesartan compared with healthy volunteers. In patients with mild to moderate or severe renal impairment there were significant increases in the maximum plasma concentration, area under the plasma drug concentration-time curve and elimination half-life of candesartan and its inactive metabolite (CV-15959) when compared to volunteers with normal renal function following repeated administration of candesartan cilexetil 8 or 12 mg. However, there was no evidence of accumulation following treatment with the 8 mg dose apart from those with severe disease requiring dialysis. Nevertheless, dialysis itself did not appear to affect the pharmacokinetic profile of candesartan or that of CV-15959. Candesartan cilexetil was found to have a good safety profile and to be well tolerated by patients with hepatic or renal impairment. There were no clinically relevant changes detected in vital signs, laboratory safety parameters or in ECG readings. The most common adverse events were headache and dizziness. This series of studies show that candesartan cilexetil 8 mg once daily is suitable for administration to patients with mild to moderate renal or hepatic impairment with no need for additional dose adjustment. A lower starting dose may be appropriate in patients with severe renal impairment including those requiring dialysis. PMID- 9331005 TI - Effects of an acute dose of 16 mg candesartan cilexetil on systemic and renal haemodynamics in hypertensive patients. PMID- 9331006 TI - Inhibition of angiotensin II pressor response and ex vivo angiotensin II radioligand binding by candesartan cilexetil and losartan in healthy human volunteers. PMID- 9331007 TI - Candesartan cilexetil, a new generation angiotensin II antagonist, provides dose dependent antihypertensive effect. AB - Candesartan is a new generation angiotensin II type 1 receptor blocker, characterised by tight binding to and slow dissociation from the receptor. In order to delineate the dose-response curve for candesartan cilexetil (the orally administered prodrug), results from six European placebo-controlled, dose response studies were pooled. These were of a double-blind, randomised, parallel group design, with a treatment duration of 4-12 weeks. A total of 1482 patients with mild to moderate primary hypertension were treated with candesartan cilexetil 2 mg (n = 80), 4 mg (n = 216), 8 mg (n = 455) or 16 mg (n = 294), or with placebo (n = 437). Blood pressure (BP) measurements were performed 24 h after dose. The differences in BP change (baseline vs end of the studies) between the placebo group and the groups treated with candesartan cilexetil were assessed using analysis of covariance and dose response curves were estimated by fitting the data to an Emax model. The placebo-corrected mean reductions in sitting diastolic BP were approximately 2.5 mm Hg with 2 mg, 4.5 mm Hg with 4 mg, 6 mm Hg with 8 mg, and 8 mm Hg with 16 mg of candesartan cilexetil. For sitting systolic BP, the placebo-corrected mean reductions were in the order of 5, 7, 10 and 12 mmHg, respectively, with 2, 4, 8 and 16 mg of candesartan cilexetil. The BP reductions were similar in the standing position with no indication of postural hypotension. Age or gender did not influence the BP response to candesartan cilexetil. In conclusion, candesartan cilexetil provides a clinically significant, dose-dependent antihypertensive effect in doses ranging from 4-16 mg once daily. PMID- 9331008 TI - Twenty-four hour blood pressure profile of different doses of candesartan cilexetil in patients with mild to moderate hypertension. PMID- 9331009 TI - Candesartan cilexetil and enalapril are of equivalent efficacy in patients with mild to moderate hypertension. PMID- 9331010 TI - Antihypertensive effects of candesartan cilexetil, enalapril and placebo. PMID- 9331011 TI - A comparison of the antihypertensive effects of candesartan cilexetil and losartan in patients with mild to moderate hypertension. PMID- 9331012 TI - Combination therapy with candesartan cilexetil plus hydrochlorothiazide in patients unresponsive to low-dose hydrochlorothiazide. PMID- 9331013 TI - Dose-finding study of candesartan cilexetil plus hydrochlorothiazide in patients with mild to moderate hypertension. PMID- 9331014 TI - Long-term efficacy and tolerability of candesartan cilexetil in patients with mild to moderate hypertension. AB - The long-term efficacy and tolerability of candesartan cilexetil was assessed in two open-label, prospective multicentre studies in patients with mild to moderate essential hypertension. When administered in a flexible dosage regimen of 4-16 mg once-daily, candesartan cilexetil effectively lowered blood pressure (BP) and maintained its antihypertensive effects over the long term (< or =12 months). At the end of treatment, 81.1% of patients showed a clinically significant response (reduction in sitting diastolic BP of > or =10 mm Hg or reduction to <90 mm Hg), and 73.8% experienced normalisation of sitting diastolic BP (<90 mm Hg). Only a small proportion (10.7%) of patients prematurely discontinued treatment due to lack of efficacy. Candesartan cilexetil was well tolerated and was devoid of clinically relevant biochemical, haematological or cardiac effects. Only 12% of adverse events were judged to be causally related to the drug and only about 5% of patients withdrew from therapy due to adverse events. The most common adverse events were typical of hypertensive patients in general. Most adverse events appeared during the first 3 months of treatment and their incidence decreased steadily with time. Tolerability was unrelated to gender, age (<65 vs > or =65 years) or dosage. These results demonstrate that candesartan cilexetil maintains its antihypertensive effects and tolerability during long-term administration. PMID- 9331015 TI - The efficacy and tolerability of candesartan cilexetil in an elderly hypertensive population. AB - This study was performed to evaluate the antihypertensive efficacy and tolerability of candesartan cilexetil 8-16 mg once-daily in comparison with placebo in elderly hypertensive patients. Forty-one hospital and general practice centres in the Netherlands and in the United Kingdom enrolled 350 patients over 65 years of age with essential hypertension (WHO grades I or II). Patients with supine diastolic BP in the range 95-114 mm Hg after 4- to 8-week placebo run-in period (n = 193) were randomised to double-blind therapy with candesartan cilexetil or placebo. The initial dose of candesartan cilexetil 8 mg or placebo was doubled after 6 weeks if supine diastolic blood pressure (BP) exceeded 90 mm Hg. Mean (95% confidence interval) placebo-corrected reduction in supine diastolic BP after 12 weeks' treatment with candesartan cilexetil was 7.5 mm Hg (3.6-11.4; P < 0.001); the corresponding reduction in supine systolic BP was 13.6 mm Hg (6.9-20.2; P < 0.001). Placebo-corrected mean reduction in supine diastolic BP 2 and 4 h after the first dose of candesartan cilexetil were 2.2 mm Hg (-1.3 to +5.8; P = 0.219) and 4.0 mm Hg (-0.4 to +7.6; P = 0.027), respectively. Candesartan cilexetil had almost no influence on heart rate and did not affect the normal orthostatic changes in BP. Adverse events were equally common in the two treatment groups. Candesartan cilexetil 8-16 mg once-daily is an effective antihypertensive agent in elderly patients. The onset of action is smooth with no exaggerated response after the first dose and there is no postural hypotension. Candesartan cilexetil is very well tolerated in elderly hypertensives. PMID- 9331016 TI - Long-term treatment with candesartan cilexetil does not affect glucose homeostasis or serum lipid profile in mild hypertensives with type II diabetes. PMID- 9331017 TI - Candesartan cilexetil: safety and tolerability in healthy volunteers and patients with hypertension. AB - The tolerability and safety of candesartan cilexetil has been evaluated in over 5000 subjects enrolled into double-blind or open-label clinical studies. In double-blind clinical trials in patients with primary hypertension, candesartan cilexetil 2-16 mg once-daily was associated with a low incidence of adverse events and drug-related withdrawals, similar to placebo. The drug showed no evidence of dose-dependent adverse events and it was equally well tolerated by men and women and by elderly (> or =65 years) and younger (<65 years) patients alike. Candesartan cilexetil had no effect on blood glucose control or serum lipid profile in patients with type II diabetes. It was very well tolerated also when given in combination with hydrochlorothiazide or amlodipine and during long term open-label therapy (up to 1 year). Candesartan cilexetil therefore possesses an excellent tolerability profile that extends to a wide variety of patients including the elderly and it does not aggravate co-existing risk factors such as hyperlipidaemia or glucose intolerance. It therefore appears to offer a better tolerated alternative to other commonly used antihypertensive agents. PMID- 9331018 TI - Candesartan cilexetil: a new, long-acting, effective angiotensin II type 1 receptor blocker. AB - Candesartan is a potent and selective angiotensin II type 1 receptor blocker which binds tightly to and dissociates slowly from the AT1 receptor. It is administered orally as the prodrug candesartan cilexetil, which is rapidly and completely converted to the active compound, candesartan, during gastrointestinal absorption. In hypertensive patients, candesartan cilexetil dose-dependently lowers diastolic and systolic blood pressure (BP) over the 24-h dose interval and it maintains its antihypertensive effects in the long term. Clinical trials indicate that a once-daily dose of 4-16 mg provides a clinically relevant reduction in BP. The usual maintenance doses of candesartan cilexetil are expected to be 8 mg and 16 mg once-daily and dosage adjustment does not appear to be necessary in elderly patients or those with mild to moderate renal or hepatic impairment. Adverse events during treatment with candesartan cilexetil occur at a similar low incidence as with placebo and no dose-dependent events or adverse metabolic effects have been noted. As once-daily monotherapy, candesartan cilexetil 8 mg is as effective as enalapril 10-20 mg, amlodipine 5 mg or hydrochlorothiazide 25 mg, and candesartan cilexetil 16 mg is more effective than losartan 50 mg. The trough to peak ratio for the reduction in BP with candesartan cilexetil has been shown to be in the order of 80-100%, confirming the smooth 24 h BP-lowering profile of the drug. Combined treatment with candesartan cilexetil and hydrochlorothiazide or amlodipine provides an enhanced BP-lowering effect that is useful in patients with inadequate response to initial treatment after step-up titration. Candesartan cilexetil is similarly well tolerated as placebo, both when given as monotherapy, and in combination for example with hydrochlorothiazide. Candesartan cilexetil with its flexible dosage regimen therefore appears to offer an effective and well-tolerated alternative to other established agents in the treatment of a wide range of hypertensive patients. PMID- 9331019 TI - Taxonomy and treatment--a classification of fracture classifications. PMID- 9331020 TI - Treatment of developmental dislocation of the hip in children after walking age. Indications from two-directional arthrography. AB - We treated 120 children between the ages of 12 and 31 months with 137 developmental dislocations of the hip and reviewed them at a mean follow-up of 14 years. We had used two-directional arthrography of all hips before reduction to evaluate the anterior, superior, and posterior portions of the limbus. Of the 137 hips, 64 had no interposed limbus in the AP view of the arthrogram, but 45 of these had an interposed anterior or posterior portion of the limbus. The hips with good stability and no interposed limbus in either AP or lateral arthrograms had excellent results by closed methods; in the other cases the results were less satisfactory. Our findings suggest that hips suitable for management by closed reduction can be identified by two-directional arthrography. Hips shown to have an interposed limbus are best managed by open reduction. PMID- 9331021 TI - Monitoring the treatment of developmental dysplasia of the hip with the Pavlik harness. The role of ultrasound. AB - We report the six-year results of a prospective, controlled demographic trial of developmental dysplasia of the hip (DDH) treated in the Pavlik harness using ultrasound supervision. Our aim was to assess the value of ultrasound and its role in monitoring reduction in the harness, in terms of progression or failure of reduction at an early state. From 1988 to 1994, a total of 221 patients with 370 ultrasonographically abnormal hips was treated in the Pavlik harness. This represents a treatment rate for the Southampton district of 5.1 per 1000 live births. Sixteen hips in 12 patients were not reduced in the harness and required surgical treatment; 95.7% were successfully reduced. One case of mild avascular necrosis (0.3%) was identified in those treated by harness alone. Of the 221 patients 87.8% remain under radiological review, with 3.2% of affected hips showing continued, mild acetabular dysplasia. We conclude that ultrasound monitoring has led to an acceptably low level of intervention, a high reduction rate and minimal iatrogenic complications. The trial is continuing. PMID- 9331022 TI - MRI after operative reduction for developmental dysplasia of the hip. AB - We performed MRI on 13 infants after operative reduction for developmental dysplasia of the hip (DDH). Using an axial gradient-echo sequence, MRI accurately depicted the acetabular anatomy and confirmed adequate reduction in 12 patients. The one patient with redislocation after surgery was correctly identified. MRI can be carried out quickly, inexpensively and without risk of radiation and is the investigation of choice to confirm adequate reduction in DDH. PMID- 9331023 TI - Incomplete healing of simple bone cysts after steroid injections. AB - We reviewed 32 children after the treatment of simple bone cysts by intralesional injections of methylprednisolone acetate. The age of the child and the activity and size of the cyst did not significantly affect the radiological outcome. The earliest time at which the radiological response could be reliably determined was three months. After a median period of review of five years, four (13%) cysts had healed, 20 (62%) cysts were partially visible but sclerotic, four (12.5%) were still visible but opaque and four (12.5%) were clearly visible. The healed and partially visible but sclerotic cysts were classified as having satisfactory radiological healing. This was observed in 13 of 32 cysts (41%) after the first injection, in eight of 21 (38%) after the second injection, but in relatively few of the remaining cysts after subsequent injections. A satisfactory symptomatic outcome was achieved in all of the 18 children with humeral cysts and in the one child with a fibular cyst irrespective of the radiological outcome, but only in nine (67%) of the 13 children with femoral or tibial lesions, in whom the cysts were healed or sclerotic. The remaining four children had exertional bone pain and repeated fractures of their femoral or tibial cysts which were incompletely healed with sclerosis in one and opacities in three. We conclude that the healing response to intralesional corticosteroids is unpredictable and usually incomplete even after multiple injections. The failure rate in weight-bearing bones is too high. PMID- 9331024 TI - Intraoperative arthrography and the Ilizarov technique. Role in the correction of paediatric deformity and leg lengthening. AB - We performed intraoperative arthrography of the knee in 12 children with congenital short femur, Blount's disease or Ollier's disease in whom the Ilizarov technique was used for correction of deformity, leg lengthening or both. In each case, arthrography revealed a joint surface considerably different from that assumed from plain radiographs, and resulted in a change in the placement of our reference wires before application of the frame. This gave significant improvement in the mechanical axis obtained at the time of removal of the frame. The technique is safe, cheap and easy to perform. It is a useful adjunct to the application of the Ilizarov frame when used for complex lengthening and correction of deformity in the leg. PMID- 9331025 TI - A modified Kapandji procedure for Smith's fracture in children. AB - Anteriorly displaced fractures of the wrist can be treated by the Kapandji technique of percutaneous intrafocal pinning with pins inserted through an anterior approach to give good reduction and stabilisation of the fracture. We have modified this technique by placing the pins through a posterior approach which decreases the risks of neurovascular damage. We have used this method to treat six children with distal radial fractures showing anterior displacement or instability. Good anterior stabilisation was achieved. The pins were removed at an average of eight weeks and the patients were then able to return to full activity. This simple technique can be used for unstable fractures after the failure of conservative treatment or in bilateral fractures in adolescents. PMID- 9331026 TI - Proximal thigh pain after femoral nailing. Causes and treatment. AB - We have reviewed retrospectively 80 patients who were treated for traumatic fractures of the femur with a Grosse-Kempf nail to assess the incidence and causes of persisting pain in the proximal thigh. At a mean of 21 months after operation 33 patients had residual pain severe enough to interfere with their lifestyle or mobility. This was in the region of the scar on the greater trochanter in three-quarters of the patients. Only four showed no radiological abnormality. There was nonunion of the fracture in two, Paget's disease in one, breakage of the nail in two and prominence of the proximal locking screw in five, although we found no correlation between prominence of the nail and pain. There was a strong relationship between pain and heterotopic ossification at the proximal end of the implant; this was present in 64% of the patients with pain as compared with those without pain (p < 0.001, Mann-Whitney U test). Of the 80 patients, 27 had the implant removed after 18 months, 17 of them because of pain. In six of these 17, the pain was not relieved. Prominence of the nail proximally was not associated with pain, but protuberance of laterally-based proximal locking screws caused problems. We found a strong association between heterotopic bone formation and pain, but it is uncertain whether this is the true cause or merely an indication of some other factor such as traumatic damage to the glutei during insertion of the nail. Removal of the implant does not always cure such pain. PMID- 9331027 TI - Charcot arthropathy after acetabular fracture. AB - Three middle-aged patients with diabetes sustained fractures of the acetabulum which were treated by open reduction and internal fixation. In each, rapid dissolution of the femoral head occurred with minimal discomfort, typical of a Charcot arthropathy. The patients had no other evidence of neuropathic arthropathy. Charcot changes may occur after high-energy trauma in patients with diabetes. PMID- 9331028 TI - Regeneration of bone after loss of the distal half of the humerus. Case report with a 20-year follow-up. AB - A 16-year-old boy was involved in an agricultural accident in which he sustained a large wound to the right arm and forearm. Radiological examination showed loss of the distal half of the humerus. A posterior splint was applied and after two months there was regeneration of the distal humerus including the articular portion. He was able to use his arm at five months. Twenty years later, he had a painless elbow and a 70 degrees range of movement. PMID- 9331029 TI - Supracondylar osteotomy with Ilizarov fixation for elbow deformities in adults. AB - Stable fixation after a corrective supracondylar osteotomy in adults is difficult because of the irregularity of the area of bony contact, displacement of the fragments, the predominance of cortical bone, and the need for early mobilisation. We have used the Ilizarov apparatus for fixation in 15 patients who were treated by complex osteotomies with displacement of fragments for cubitus varus or valgus. Most patients with cubitus varus required medial displacement with rotation of the distal fragment. Those with cubitus valgus required lateral shift of the distal fragment to reduce the medial prominence of the elbow that would otherwise result. All osteotomies united within the expected time without loss of correction, despite early mobilisation. Complications related to the fixation were few and had resolved at the long-term follow-up. PMID- 9331030 TI - Wrist involvement in Hansen's disease. AB - We performed a neurological and radiological study of the wrists of 58 patients with Hansen's disease and 60 age-matched healthy control subjects. Significant differences (p < 0.01) were found between the groups in the carpal glenoid sector, the radial physeal widening index, the carpal ulnar distance, the carpal index and in distal radio-ulnar discrepancy. Comparison of the results in three subgroups of leprous patients with sensory impairment (group A-1), motor deficit (A-2) and no neurological impairment (A-3), showed significant differences (p < 0.01) between group A-1 and the other two. This suggests that in these patients the changes in the carpus and radiocarpal joint may be caused by neuropathic arthropathy of the wrist. Our findings are of particular interest since there are few reports of neuropathic arthropathy in non-weight-bearing joints. PMID- 9331031 TI - Fractures of the base of the middle phalanx of the finger. Classification, management and long-term results. AB - We classified fractures of the base of the middle phalanx into five types: 1) single palmar fragment; 2) single dorsal fragment; 3) two main fragments; 4) not involving the articular surface, including epiphyseal separation in children; and 5) all others. Types 1 and 2 were subclassified into avulsion, split and split depression. Surgery is recommended for unstable type-1 avulsion fractures, type-2 avulsions which may develop buttonhole deformities, and all fractures which displace articular cartilage surfaces. Long-term follow-up showed that surgical treatment which produced good stability and congruity gave good results. These should be the primary aims of treatment. PMID- 9331032 TI - Magnetic resonance myelography in brachial plexus injury. AB - We used magnetic resonance (MR) myelography in ten patients with injuries to the brachial plexus and compared the findings with those obtained by conventional myelography and postmyelographic CT (CTM). In the presence of complete nerve-root avulsion (seven cases), a post-traumatic meningocele was detected by MR myelography. In injuries to the upper roots (three cases) MR myelography showed abnormal findings with a high signal intensity in the nerve root, obliteration of the damaged nerve root, or enlargement and obliteration of the root sleeve. No pseudomeningoceles were detected in these upper-root injuries by MR myelography and CTM. The overall accuracy of detection of damaged nerve roots or root sleeves was better with MR myelography than with conventional myelography and was similar to that of CTM. MR myelography is non-invasive, relatively quick, requires no contrast medium, provides imaging in multiple projections, and is comparable in diagnostic ability to the more invasive, time-consuming techniques of conventional myelography and CTM. PMID- 9331033 TI - Arthroscopic synovectomy of the elbow for rheumatoid arthritis. A prospective study. AB - The short-term assessment of 14 arthroscopic synovectomies of the elbow in 11 patients with rheumatoid arthritis showed that 93% achieved a short-term rating of excellent or good on the Mayo Elbow Performance Score. At the most recent assessment at an average of 42 months, however, only 57% maintained excellent or good results; four had required total elbow replacement. Although rehabilitation is facilitated by an arthroscopic procedure the results deteriorate more rapidly than after open synovectomy. This may be due to the limitations of the arthroscopic technique and is consistent with experience of the similar procedure in the knee. Recognition of the short-term gain and the potential for serious nerve injury should be considered when offering arthroscopic synovectomy. PMID- 9331034 TI - Reconstruction of the hemipelvis after the excision of malignant tumours. Complications and functional outcome of prostheses. AB - We treated 35 patients with primary malignant tumours of the periacetabular area by resection and prosthetic reconstruction of the defect. At a mean follow-up of 84 months, 15 patients (43%) were free from disease. The most common complications were deep infection (26%), local recurrence (24%) and recurrent dislocation of the hip (17%). The surviving patients achieved an average of 70% of their premorbid function. This method of reconstruction has a high morbidity and should be performed only at specialist centres, but the functional and oncological outcomes are satisfactory. PMID- 9331035 TI - Symptomatic venous thromboembolism after total knee replacement. AB - Chemical prophylaxis is known to reduce the venographic prevalence of deep-vein thrombosis (DVT) after total knee replacement (TKR), but it is uncertain whether this affects the incidence of symptoms. Further analysis depends on the basic epidemiology of thromboembolic symptoms. We therefore studied the pattern of such symptoms in a consecutive series of 1000 patients with primary TKR, with particular reference to risk factors and prophylaxis. We reviewed all the clinical records and contacted all the patients individually, noting risk factors, prophylaxis, symptomatic pulmonary embolus (PE) or DVT and its timing, death and its causes, and all complications. All the patients wore antiembolism stockings, 83% had regional anaesthesia and 33.9% had chemical prophylaxis. One patient died from PE on the day of surgery, having had no prophylaxis giving a rate of 0.1% (95% CI 0.003% to 0.56%). Symptomatic, radiologically confirmed thromboembolism (VTE) was common with a rate of 10.6% (95% CI 8.7% to 12.5%). There was a similar incidence of VTE in those with and without chemical prophylaxis (10.1% v 10.5%, RR 0.96, NS). VTE was more common in patients with risk factors (15.1% v 9.5%, RR 1.59, p = 0.02) and tended to occur earlier in this group (median day of onset 5 v 7, p = 0.01). Chemical prophylaxis did not reduce the frequency of symptomatic thromboembolism in either those with risk factors (RR 0.81, p = 0.5) or those without them (RR 0.94, p = 0.8). Haematoma or wound dehiscence was more common in those having chemical prophylaxis (11.9% v 6.9%; RR 1.73 95% CI 1.16 to 2.60). Readmission for symptomatic, radio-logically confirmed thromboembolism involved 1.1% of patients (95% CI 0.55% to 2.1%). Four patients were readmitted with proven non-fatal PE and six with proven DVT (the latest on day 40). Our results show that the main risk factor for thromboembolism was TKR itself; chemical prophylaxis did not reduce the incidence of symptomatic thromboembolism but gave an increased perception of side-effects. New prophylactic methods or combinations of methods are needed, with their efficacy compared by randomised controlled studies of both the clinical and the radiological effect. PMID- 9331036 TI - Screening for deep-venous thrombosis after hip and knee replacement without prophylaxis. AB - We performed routine venography after operation in a consecutive series of 252 patients with total joint arthroplasties in whom no form of routine chemical or mechanical prophylaxis had been used. The prevalence of deep-vein thrombosis (DVT) was 32% (16% distal, 16% proximal) after total hip replacement and 66% (50% distal, 16% proximal) after total knee replacement (p < 0.001). We did not treat distal DVT. There were only two readmissions within three months of surgery because of thromboembolic disease. There were two deaths within this period, neither of which was due to pulmonary embolism. PMID- 9331037 TI - Survivorship of the Charnley total hip arthroplasty in juvenile chronic arthritis. A follow-up of 186 cases for 22 years. AB - Between 1971 and 1991 we performed Charnley low-friction arthroplasty (LFA) on 116 patients (186 hips) with juvenile chronic arthritis (JCA). We have now carried out a survival study, taking endpoints as revision, death or the end of the year 1993. Overall survival was 91.9% at ten years and 83.0% at 15 years. That of the femoral component was 95.6% at ten years and 91.9% at 15 years and of the acetabulum 95.0% and 87.8%, respectively. Only the use of steroids significantly impaired the survival. We therefore recommend the use of Charnley LFA for young patients with JCA requiring total hip replacement. PMID- 9331038 TI - Intra-articular local anaesthesia for pain after hip arthroplasty. AB - We investigated 15 patients with painful hip arthroplasties using intra-articular injection of bupivicaine. Fourteen had pain relief and 13 of them were subsequently found to have loosening of one or both components. The relief of pain after total hip arthroplasty by intra-articular injection of bupivicaine indicates that a satisfactory result is probable after revision surgery with refixation of the components. PMID- 9331039 TI - The superficial peroneal tunnel syndrome. Results of treatment by decompression. AB - We diagnosed entrapment of the superficial peroneal nerve in 17 patients (19 legs) with a mean age of 41 years. In all cases, plain radiographs of the leg, nerve-conduction studies of the superficial peroneal nerve and measurement of the intramuscular pressure at rest after exercise were normal. Diagnostic tests for nerve compression during rest after exercise produced pain and clinical signs in all. We performed decompression of the superficial peroneal tunnel in 14 patients and local fasciectomy in three. Fourteen patients (80%) were free from symptoms or satisfied with the result. PMID- 9331040 TI - Foraminal injection for lateral lumbar disc herniation. AB - Between 1986 and 1995, we treated with foraminal injection of local anaesthetic and steroids 30 patients with severe lumbar radiculopathy secondary to foraminal and extraforaminal disc herniation which had not resolved with rest and non steroidal anti-inflammatory agents. They were assessed prospectively using standardised forms as well as the Low Back Outcome Score, and were reviewed at an average of 3.4 years (1 to 10) after injection by an independent observer (BKW). Relief of symptoms was obtained in 27 immediately after injection. Three subsequently relapsed, requiring operation, and two were lost to long-term follow up. Thus 22 of the 28 patients available for long-term follow-up had considerable and sustained relief from their symptoms. Before the onset of symptoms 17 were in employment and, after injection, 13 resumed work, all but two in the same job. The average score before injection was 25 out of a possible 75 points. At follow up, the overall average score was 54, and in those who had obtained relief of symptoms it had improved to a mean of 61. Based on these findings we recommend foraminal injection of local anaesthetic and steroids as the primary treatment for patients with severe radiculopathy secondary to foraminal or extraforaminal herniation of a lumbar disc. PMID- 9331041 TI - Preoperative endovascular embolisation of a vertebral haemangioma. AB - We describe the successful relief of compression of the spinal cord due to a vertebral haemangioma by transcatheter embolisation using cyanoacrylate compounds before operation, and provide a brief review of the literature. PMID- 9331042 TI - Solitary plasma-cell myeloma of the spine in an adolescent. Case report of an unusual presentation. AB - We report an unusual presentation of a solitary plasma-cell myeloma of the spine in an adolescent patient. Our case indicates the need to consider plasma-cell myeloma as a differential diagnosis even in younger patients. PMID- 9331043 TI - The prevention of prosthetic infection using a cross-linked albumin coating in a rabbit model. AB - We evaluated the effects of a serum protein coating on prosthetic infection in 29 adult male rabbits divided into three groups: control, albumin-coated and uncoated. We used 34 grit-blasted, commercially pure titanium implants. Eleven were coated with cross-linked albumin. All the implants were exposed to a suspension of Staphylococcus epidermidis before implantation. Our findings showed that albumin-coated implants had a much lower infection rate (27%) than the uncoated implants (62%). This may be a useful method of reducing the infection of prostheses. PMID- 9331044 TI - Variation of the groove in the axis vertebra for the vertebral artery. Implications for instrumentation. AB - Transarticular screws at the C1 to C2 level of the cervical spine provide rigid fixation, but there is a danger of injury to a vertebral artery. The risk is related to the technical skill of the surgeon and to variations in local anatomy. We studied the grooves for the vertebral artery in 50 dry specimens of the second cervical vertebra (C2). They were often asymmetrical, and in 11 specimens one of the grooves was deep enough to reduce the internal height of the lateral mass at the point of fixation to < or =2.1 mm, and the width of the pedicle on the inferior surface of C2 to < or =2 mm. In such specimens, the placement of a transarticular screw would put the vertebral artery at extreme risk, and there is not enough bone to allow adequate fixation. Before any decision is made concerning the type of fixation to be used at C2 we recommend that a thin CT section be made at the appropriate angle to show both the depth and any asymmetry of the grooves for the vertebral artery. PMID- 9331045 TI - Three modes of ossification during distraction osteogenesis in the rat. AB - We developed a rat model of limb lengthening to study the basic mechanism of distraction osteogenesis, using a small monolateral external fixator. In 11-week old male rats we performed a subperiosteal osteotomy in the midshaft of the femur with distraction at 0.25 mm every 12 hours from seven days after operation. Radiological and histological examinations showed a growth zone of constant thickness in the middle of the lengthened segment, with formation of new bone at its proximal and distal ends. Osteogenic cells were arranged longitudinally along the tension vector showing the origin and the fate of individual cells in a single section. Typical endochondral bone formation was prominent in the early stage of distraction, but intramembraneous bone formation became the predominant mechanism of ossification at later stages. We also showed a third mechanism of ossification, 'transchondroid bone formation'. Chondroid bone, a tissue intermediate between bone and cartilage, was formed directly by chondrocyte-like cells, with transition from fibrous tissue to bone occurring gradually and consecutively without capillary invasion. In situ hybridisation using digoxigenin 11-UTP-labelled complementary RNAs showed that the chondroid bone cells temporarily expressed type-II collagen mRNA. They did not show the classical morphological characteristics of chondrocytes, but were assumed to be young chondrocytes undergoing further differentiation into bone-forming cells. We found at least three different modes of ossification during bone lengthening by distraction osteogenesis. We believe that this is the first report of such a rat model, and have shown the validity of in situ hybridisation techniques for the study of the cellular and molecular mechanisms involved in distraction osteogenesis. PMID- 9331046 TI - Chondrocyte transplantation using a collagen bilayer matrix for cartilage repair. AB - We have developed a novel, two-layered, collagen matrix seeded with chondrocytes for repair of articular cartilage. It consists of a dense collagen layer which is in contact with bone and a porous matrix to support the seeded chondrocytes. The matrices were implanted in rabbit femoral trochleas for up to 24 weeks. The control groups received either a matrix without cells or no implant. The best histological repair was seen with cell-seeded implants. The permeability and glycosaminoglycan content of both implant groups were nearly normal, but were significantly less in tissue from empty defects. The type-II collagen content of the seeded implants was normal. For unseeded implants it was 74.3% of the normal and for empty defects only 20%. The current treatments for articular injury often result in a fibrous repair which deteriorates with time. This bilayer implant allowed sustained hyaline-like repair of articular defects during the entire six month period of observation. PMID- 9331047 TI - Benign cellular responses in rats to different wear particles in intra-articular and intramedullary environments. AB - We examined the cellular responses to various particles injected into the knees and the intramedullary femoral cavities of rats in the presence of polymethylmethacrylate (PMMA) plugs. The intra-articular particles were mainly ingested by synovial fibroblasts. Increased numbers of macrophages were not detected and there was only a slight increase in synovial thickness. Cellular responses in the intramedullary space were similarly mild and bone resorption around the PMMA plug did not occur. Bone formation was inhibited only by polyethylene particles. In contrast to current views, our study shows that wear particles per se do not initiate bone resorption. PMID- 9331048 TI - Number of polyethylene particles and osteolysis in total joint replacements. A quantitative study using a tissue-digestion method. AB - Our aim was to analyse the influence of the size, shape and number of particles on the pathogenesis of osteolysis. We obtained peri-implant tissues from 18 patients having revision surgery for aseptically loosened Freeman total knee replacements (10), Charnley total hip replacements (3) and Imperial College/London Hospital double-cup surface hip replacements (5). The size and shape of the polyethylene particles were characterised using SEM and their concentration was calculated. The results were analysed with reference to the presence of radiological osteolysis. The concentration of polyethylene particles in 6 areas with osteolysis was significantly higher than that in 12 areas without osteolysis. There were no significant differences between the size and shape of the particles in these two groups. We conclude that the most critical factor in the pathogenesis of osteolysis is the concentration of polyethylene particles accumulated in the tissue. PMID- 9331049 TI - Biomaterial particle phagocytosis by bone-resorbing osteoclasts. AB - Abundant implant-derived biomaterial wear particles are generated in aseptic loosening and are deposited in periprosthetic tissues in which they are phagocytosed by mononuclear and multinucleated macrophage-like cells. It has been stated that the multinucleated cells which contain wear particles are not bone resorbing osteoclasts. To investigate the validity of this claim we isolated human osteoclasts from giant-cell tumours of bone and rat osteoclasts from long bones. These were cultured on glass coverslips and on cortical bone slices in the presence of particles of latex, PMMA and titanium. Osteoclast phagocytosis of these particle types was shown by light microscopy, energy-dispersive X-ray analysis and SEM. Giant cells containing phagocytosed particles were seen to be associated with the formation of resorption lacunae. Osteoclasts containing particles were also calcitonin-receptor-positive and showed an inhibitory response to calcitonin. Our findings demonstrate that osteoclasts are capable of phagocytosing particles of a wide range of size, including particles of polymeric and metallic biomaterials found in periprosthetic tissues, and that after particle phagocytosis, they remain fully functional, hormone-responsive, bone resorbing cells. PMID- 9331050 TI - The three-dimensional geometry of the proximal humerus. Implications for surgical technique and prosthetic design. AB - We have studied the three-dimensional geometry of the proximal humerus on human cadaver specimens using a digitised measuring device linked to a computer. Our findings demonstrated the variable shape of the proximal humerus as well as its variable dimensions. The articular surface, which is part of a sphere varies individually in its orientation as regards inclination and retroversion, and it has variable medial and posterior offsets. These variations cannot be accommodated by the designs of most contemporary humeral components. Although good clinical results can be achieved with current modular and non-modular components their relatively fixed geometry prevents truly anatomical restoration in many cases. To try to restore the original three-dimensional geometry of the proximal humerus, we have developed a new type of humeral component which is modular and adaptable to the individual anatomy. Such adaptability allows correct positioning of the prosthetic head in relation to an individual anatomical neck, after removal of the marginal osteophytes. The design of this third-generation prosthesis respects the four geometrical variations which have been demonstrated in the present study. These are inclination, retroversion, medial offset and posterior offset. PMID- 9331051 TI - Alternative treatments for meniscal injuries. PMID- 9331052 TI - Femoral component revision using impacted morsellised cancellous graft. PMID- 9331053 TI - Ludloff's medial approach for open reduction of congenital dislocation of the hip. PMID- 9331054 TI - Ludloff's medial approach for open reduction of congenital dislocation of the hip. PMID- 9331055 TI - Brachial plexus injuries. PMID- 9331056 TI - Simple bone cysts treated by injection of autologous bone marrow. PMID- 9331057 TI - Simple bone cysts treated by injection of autologous bone marrow. PMID- 9331058 TI - Care of the polytraumatised patient. PMID- 9331059 TI - Survival analysis of joint replacements. PMID- 9331060 TI - Thromboprophylaxis and death after total hip replacement. PMID- 9331061 TI - Thromboprophylaxis and death after total hip replacement. PMID- 9331062 TI - Thromboprophylaxis and death after total hip replacement. PMID- 9331063 TI - Which primary total hip replacement. PMID- 9331064 TI - Characterization of a 4-Mb region at chromosome 6q21 harboring a replicative senescence gene. AB - A 4-Mb region containing a senescence gene was defined at 6q21 by fluorescence in situ hybridization and deletion mapping after transfer of a normal human chromosome 6 to a BK virus-transformed mouse cell line. By screening three different yeast artificial chromosome (YAC) libraries, a YAC contig was constructed that covers the deleted region at 6q21. The contig is composed of 18 overlapping YACs with a size of 250-1800 kb and contains 3 CpG islands and 10 expressed sequence tags. By sequencing YACs and P1 artificial chromosomes, nine new sequence tagged sites and three new expressed sequence tags were detected that enrich the genetic resources of the region. The contig may also contain a fragile site, FRA6F, located close to a CpG island, which could be a landmark to localize the senescence gene. This YAC contig will be used to detect expressed sequences to clone and characterize the senescence gene at 6q21. PMID- 9331065 TI - Loss of retinoic acid receptor beta expression in breast cancer and morphologically normal adjacent tissue but not in the normal breast tissue distant from the cancer. AB - Retinoids and their receptors [retinoic acid receptors (RARs) and retinoid X receptors] play an important role in maintaining the balance between proliferation and apoptosis. Recently, Deng et al. [Science (Washington DC), 274: 2057-2059, 1996] reported a loss of heterozygosity on chromosome 3p24 in breast cancer specimens and the morphologically normal appearing adjacent tissue. The 3p24 locus includes, among other genes, the region coding for RAR-beta. This study was designed to determine whether there are abnormalities in the expression of retinoid receptors in surgical specimens of patients with breast cancer. In 14 patients, transcripts of nuclear retinoid receptors were detected by in situ hybridization in formalin-fixed, paraffin-embedded specimens by means of digoxigenin-labeled riboprobes specific for RAR-alpha, -beta and -gamma. We found RAR-alpha expressed in all specimens, whereas RAR-gamma was expressed in 100% of normal breast tissue but in only 11 of 14 tumorous lesions. RAR-beta was found in all cases of normal breast tissue localized distant from the tumor, but in 13 of 14 cases it was completely absent in the tumor and the morphologically normal appearing tissue adjacent to the tumor. One possibility to explain the suppression of RAR-beta is a mutation in the promoter region. Sequencing the DNA extracted from paraffin-embedded tumor tissue of the corresponding breast cancer specimens, we were not able to detect any mutation in the retinoic acid responsive element. Our results clearly indicate a crucial role of RAR-beta in the carcinogenesis of breast cancer. PMID- 9331066 TI - Multigenerational effects of dietary fat carcinogenesis in mice. AB - The possibility of multigenerational transmission of a carcinogenic effect from exposure to a maternal diet high in fat was tested in mice. Diets with 2.6 or 29% fat (by weight) were fed to strain CD-1 mice during pregnancy. The female offspring were raised on a control diet (10% fat), mated, and continued on the control diet through pregnancy. Their female offspring were raised to terminal illness and autopsied. The total number of reproductive system tumors, pituitary tumors, and metastases was increased in the offspring with ancestral exposure to high dietary fat but to a lesser extent than had been reported previously for direct prenatal exposure to high maternal dietary fat. Because previous work has given evidence against germ cell transmission, a hypothesis based on a maternal effect was offered to explain the multigenerational carcinogenesis. These results have implications for epidemiological studies. PMID- 9331067 TI - Genetic mapping of lung cancer modifier loci specifically affecting tumor initiation and progression. AB - Mouse inbred strains with inherited predisposition and resistance to lung cancer provide a tool for the dissection of the complex genetics of this disease. In the present report, we have crossed the BALB/c with the SWR/J strain and performed whole-genome scanning for loci affecting lung tumor development in their F2 progeny. Both parental strains carry the pulmonary adenoma susceptibility 1 (Pas1) locus, a major locus affecting predisposition to lung cancer in mice. On distal chromosome 18 and on centromere of chromosome 6, we have mapped two pulmonary adenoma resistance loci (Par2 and Par4, respectively), which reduce lung tumor multiplicity strongly, up to 15-fold. Par2 and Par4, however, do not affect lung tumor size, which is instead controlled by an additional locus that we have mapped on the central region of chromosome 4. We designated this locus as "pulmonary adenoma progression 1" (Papg1), because it specifically modifies lung tumor size but not multiplicity. The present results, therefore, provide evidence for the existence of cancer modifier loci acting on specific stages of lung tumorigenesis. PMID- 9331068 TI - Detection of prostate cancer cells circulating in peripheral blood by reverse transcription-PCR for hKLK2. AB - Two of the human tissue kallikrein family, hK2 and hK3 (prostate-specific antigen), are primarily produced by the prostatic epithelium under the regulation of androgens. In this study, we detected prostate cancer cells that expressed hKLK2 or hKLK3 mRNA in the peripheral blood of patients with prostate cancer using reverse transcription-PCR (RT-PCR). We then demonstrated some differences in characteristics, such as differentiation of cancer cells and response to antiandrogen therapy, between hKLK2 and hKLK3 mRNA-expressing prostate cancer cells. Total RNA was isolated from 41 patients with known prostate cancer, 7 patients with benign prostatic hyperplasia, and 20 normal volunteers. By RT-PCR, hKLK2 mRNA was detected in 7 patients (33%), and hKLK3 mRNA was detected in 17 (81%) of 21 stage D prostate cancer patients. In contrast, all patients with benign prostatic hyperplasia and healthy volunteers were negative. From comparison of the background of the patients positive for hKLK2 and/or hKLK3 mRNA, it became evident that the response to antiandrogen therapy and the expression of hKLK2 mRNA were reciprocally correlated, in contrast with the expression of hKLK3 mRNA. Additionally, our study clearly demonstrated that the detection of hKLK2 mRNA in the peripheral blood was useful for screening patients with certain prostate cancers that did not express hK3. We conclude that taking advantage of the difference between hKLK2 mRNA and hKLK3 mRNA expression is clinically useful for following up prostate cancer patients. PMID- 9331069 TI - Genetic alterations accumulate during cervical tumorigenesis and indicate a common origin for multifocal lesions. AB - Carcinomas of the uterine cervix are thought to arise from preinvasive dysplastic lesions, termed cervical intraepithelial neoplasias (CIN), grades I-III. Patients may present clinically with two or more distinct lesions of differing histological severity; however, the genesis of these multifocal lesions is unknown. Despite infection with high-risk human papilloma virus subtypes, which is a major etiological factor in disease pathogenesis, only a small and unpredictable number of dysplastic lesions progress to invasive cancer. Several lines of evidence suggest that additional somatic events, such as tumor suppressor gene inactivation, are required for malignant transformation. In support of this, loss of heterozygosity (LOH) analyses of invasive cervical carcinomas have identified several chromosomal arms likely to harbor tumor suppressor genes, of which regions on 3p, 4p, 4q, and 11q have been validated extensively. To evaluate the potential role of tumor suppressor gene inactivation in dysplastic progression, loci distributed on these four chromosomal regions were assessed for LOH in 42 CIN lesions of varying histological grade obtained from 17 patients. Analysis of at least 16 microsatellite loci in each lesion revealed allelic losses involving one or more of these chromosomal regions in 0% of CIN I lesions; 25% of CIN II lesions; and 88% of CIN III lesions, with 41% of CIN III lesions exhibiting LOH for three or more chromosomal regions. In addition, where LOH was scored for the same locus at a particular chromosomal region in all of the multiple lesions from a single patient, the same allele was lost at each locus, without exception. Statistical analysis of these allele specific losses strongly suggests that topologically distinct lesions are related and likely arise from a common precursor cell. PMID- 9331070 TI - Human mitogen-activated protein kinase kinase 4 as a candidate tumor suppressor. AB - Mitogen-activated protein kinases function in signal transduction pathways that are involved in controlling key cellular processes in many organisms. A mammalian member of this kinase family, MKK4/JNKK1/SEK1, has been reported to link upstream MEKK1 to downstream stress-activated protein kinase/JNK1 and p38 mitogen activated protein kinase. This mitogen-activated protein kinase pathway has been implicated in the signal transduction of cytokine- and stress-induced apoptosis in a variety of cell types. Here, we report that two human tumor cell lines, derived from pancreatic carcinoma and lung carcinoma, harbor homozygous deletions that eliminate coding portions of the MKK4 locus at 17p, located approximately 10 cM centromeric of p53. In addition, in a set of 88 human cancer cell lines prescreened for loss of heterozygosity, we detected two nonsense and three missense sequence variants of MKK4 in cancer cell lines derived from human pancreatic, breast, colon, and testis cells. In vitro biochemical assays revealed that, when stimulated by MEKK1, four of the five altered MKK4 proteins lacked the ability to phosphorylate stress-activated protein kinase. Thus, the incidence of coding mutations of MKK4 in the set of cell lines is 6 of 213 (approximately 3%). These findings suggest that MKK4 may function as a suppressor of tumorigenesis or metastasis in certain types of cells. PMID- 9331071 TI - Somatic mutations of PTEN in glioblastoma multiforme. AB - Alterations of the PTEN gene occur in glioblastoma multiforme. To determine the frequency of PTEN alteration, 34 consecutive glioblastomas were studied in detail. Sequencing each of the nine exons amplified from tumor DNA revealed 11 mutations. Analysis of polymorphic markers within and surrounding the PTEN gene identified an additional four homozygous deletion mutations. Loss of heterozygosity (LOH) was observed in 25 of 34 (74%) cases. All mutations occurred in the presence of LOH. PTEN was mutated in 44% (15 of 34) of all glioblastomas studied and 60% (15 of 25) of tumors with LOH on 10q. Thus, PTEN appears to be the major target of inactivation on chromosome 10q in glioblastoma multiforme. PMID- 9331072 TI - PTEN gene mutations are seen in high-grade but not in low-grade gliomas. AB - The PTEN gene, located on 10q23, has recently been implicated as a candidate tumor suppressor gene in brain, breast and prostate tumors. In the present study, 123 brain tumors, including various grades and histological types of gliomas occurring in children and adults, were analyzed for PTEN mutations by SSCP assay and sequencing. Mutations in the PTEN gene were found in 13 of 42 adult glioblastomas and 3 of 13 adult anaplastic astrocytomas, whereas none of the 21 low-grade adult gliomas or the 22 childhood gliomas of all grades showed mutations. The single medulloblastoma with a mutation was a recurrent tumor that also possessed a p53 mutation. High-grade adult gliomas with PTEN mutations included cases that also contained gene amplification or p53 gene mutations, as well as cases that did not contain either of these abnormalities. There was no obvious relationship between presence of PTEN mutation and survival; however, there was a tendency for PTEN mutations to occur in older age group patients. This analysis suggest that PTEN gene mutations are restricted to high-grade adult gliomas and that this abnormality is independent of the presence or absence of gene amplification or p53 gene mutation in these tumors. PMID- 9331073 TI - Genomic structure and genetic mapping of the human neutral cysteine protease bleomycin hydrolase. AB - Bleomycin hydrolase (BH) is the only known eukaryotic enzyme that inactivates the widely used antineoplastic agent bleomycin (BLM) and is a primary candidate gene for protection against lethal BLM-induced pulmonary fibrosis and for BLM resistance in tumors. Human BH was found to exist as a single gene that was mapped to chromosome 17 using National Institute of General Medical Sciences human/rodent hybrid mapping panels and localized to 17q11.1-11.2 by linkage analysis using the Centre d'Etude du Polymorphisme Humain reference database. The human BH gene consisted of 11 exons ranging in size from 69-198 bp separated by introns of approximately 1 kb, reflecting the archetypal genomic structure of the cysteine protease family. A polymorphic site was identified in the eleventh exon at bp 1450 encoding either valine or isoleucine. These findings provide essential tools required to define the role of BH in BLM-induced pulmonary fibrosis and BLM resistance in tumors. PMID- 9331074 TI - Pretreatment prediction of the chemotherapeutic response of human glioma cell cultures using nuclear magnetic resonance spectroscopy and artificial neural networks. AB - Both tumor metabolism and its response to cytotoxic drugs are intrinsic properties of tumor cells. It is therefore likely that there is a relationship between the two properties, however subtle and complex, wherein the metabolic characteristics of tumor cells can reflect the inherent response (resistance or sensitivity) of these cells to cytotoxic drugs. We used artificial neural network analysis to show that it is possible to distinguish, prior to treatment, between drug-resistant and drug-sensitive human glioma cell cultures from their metabolic profiles, as given by high-resolution proton nuclear magnetic resonance spectra of the cell extracts, and to predict their cellular response to the chemotherapeutic drug 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea in vitro. The results suggest that neural network analysis of tumor nuclear magnetic resonance spectra has potential as a prognostic tool for determining treatment of gliomas, ultimately noninvasively, and may be used to provide information about the metabolic pathways involved in drug response that may be helpful in developing novel treatments for these tumors. PMID- 9331075 TI - Selection of human cervical epithelial cells that possess reduced apoptotic potential to low-oxygen conditions. AB - Since human papillomavirus (HPV) infection is strongly associated with cervical neoplasia and tumor hypoxia has prognostic significance in human cervical carcinomas, we examined the relationship between hypoxia and apoptosis in human cervical epithelial cells expressing high-risk HPV type 16 oncoproteins. In vitro, hypoxia stimulated both p53 induction and apoptosis in primary cervical epithelial cells infected with the HPV E6 and E7 genes but not in cervical fibroblasts infected with E6 and E7. Furthermore, cell lines derived from HPV associated human cervical squamous cell carcinomas were substantially less sensitive to apoptosis induced by hypoxia, indicating that these cell lines have acquired additional genetic alterations that reduced their apoptotic sensitivity. Although the process of long-term cell culturing resulted in selection for subpopulations of HPV oncoprotein-expressing cervical epithelial cells with diminished apoptotic potential, the exposure of cells to hypoxia greatly accelerated the selection process. These results provide evidence for the role of hypoxia-mediated selection of cells with diminished apoptotic potential in the progression of human tumors and can in part explain why cervical tumors that possess low pO2 values are more aggressive. PMID- 9331077 TI - Basal cell tetrasomy in low-grade cervical squamous intraepithelial lesions infected with high-risk human papillomaviruses. AB - We have analyzed 60 low-grade cervical squamous intraepithelial lesions for low- and high-risk human papillomaviruses (HPVs) and for numerical abnormalities of chromosomes 1, 3, 11, 17, and 18 and the X chromosome. Eleven of 33 lesions infected with high-risk HPVs (HPV 16, 18, 30, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 66) but none of 24 lesions infected with low-risk HPVs (HPV 6, 11, 42, 43, and 44) and none of 15 normal cervices showed basal cell tetrasomy of all six chromosomes in the HPV-infected areas. These changes were not HPV type specific and were not present in all lesions infected with the same HPV type. The presence of basal cell tetrasomy in lesions infected with high- but not low-risk HPVs suggests that induction of chromosome instability may be one mechanism underlying the biological differences between these viral types. PMID- 9331076 TI - Virally directed cytosine deaminase/5-fluorocytosine gene therapy enhances radiation response in human cancer xenografts. AB - Gene therapy combined with radiation therapy to enhance selectively radiation cytotoxicity in malignant cells represents a new approach for cancer treatment. We investigated the efficacy of adenoviral (Ad5)-directed cytosine deaminase/5 fluorocytosine (CD/5-FC) enzyme/prodrug gene therapy to enhance selectively the tumoricidal action of ionizing radiation in human cancer xenografts derived from a human squamous carcinoma cell line (SQ-20B). Tumor xenografts grown in hindlimbs of nude mice were transfected with an adenoviral vector (Ad.CMV.CD) containing the cytosine deaminase (CD) gene under the control of a cytomegalovirus (CMV) promoter. Mice were injected i.p. with 800 mg/kg of 5-FC for 12 days, and tumors were treated with fractionated radiation at a dose of 5 Gy/day to a total dose of 50 Gy. In larger tumors with a mean volume of 1069 mm3, marked tumor regression to 11% of the original tumor volume was observed at day 21 (P = 0.01). The volumetric regression of smaller tumors with a mean volume of 199 mm3, which received the same combined treatment protocol, was significant at day 12 (P = 0.014). However, unlike large tumors, regression of the smaller tumors continued until day 36 (P = 0.01), with 43% cured at day 26. No cures or significant volumetric reduction in size was observed in tumors treated with radiation alone; Ad.CMV.CD with or without radiation; or with Ad.CMV.CD and 5-FC. These results suggest that the CD/5-FC gene therapy approach is an effective radiosensitizing strategy and may lead to substantial improvement in local tumor control that would translate into improved cure rates and better survival. PMID- 9331078 TI - Skin autografts in epidermodysplasia verruciformis: human papillomavirus associated cutaneous changes need over 20 years for malignant conversion. AB - Epidermodysplasia verruciformis (EV) is regarded as a model for cutaneous oncogenesis associated with specific human papillomaviruses (HPVs). Because genital HPV-associated carcinogenesis is a very long-lasting process requiring 20 30 years and epidemiological studies of this type for HPV-associated skin cancers are impossible in such a rare disease as EV, we observed for up to 20 years EV patients having surgery for carcinomas with consecutive autografts from uninvolved and non-sun-exposed skin. We noticed the appearance of premalignant and malignant changes around the grafts, whereas within the grafted skin, only benign macular lesions started to develop several years after transplantation. Thus, skin HPV-associated carcinogenesis appears to be a very slow process comparable to the genital carcinogenesis associated with high risk HPVs. PMID- 9331079 TI - A region of allelic imbalance in 1q31-32 in primary breast cancer coincides with a recombination hot spot. AB - Previous studies have shown that the 1q31-32 region frequently presents allelic imbalance (AI) in various neoplastic diseases, such as breast cancer, medulloblastoma, male germ cell tumors, and renal collecting duct carcinoma, suggesting the presence of a tumor suppressor gene in this location. We used 19 informative microsatellite markers to analyze 33 primary breast tumors for AI in the 1q31-32 region. Our results demonstrate a 10-cM critical region of AI that is present in more than 60% of the tumors. This region is located proximal to the REN locus and is flanked by the CACNL1A3 and D1S2655 markers. Most important, the critical region of AI coincides with a female hot spot of recombination, suggesting a possible correlation between the two regions. PMID- 9331080 TI - Inactivation of Smad4 in gastric carcinomas. AB - Allelic loss of chromosome 18q has been noted in intestinal type gastric adenocarcinomas. Smad4 is a gene located at 18q that was recently cloned in humans and found to be significantly altered in pancreatic cancers. We sought to determine whether Smad4 genetic alterations played a significant role in gastric tumorigenesis by studying 35 gastric adenocarcinomas of all histopathological types and pathological stages. Microdissected specimens were used for mutational analysis of Smad4 at the nucleotide level, including the entire coding region and intron/exon boundaries. Allelic imbalance was also analyzed at the Smad4 locus using two nearby microsatellite markers. One case of apparent biallelic inactivation of Smad4 was found in our study of 35 gastric carcinomas. A nonsense point mutation at codon 334 was demonstrated, which, similar to other Smad4 mutations, is predicted to truncate the conserved COOH-terminal domain of this protein. This Smad4 C to T transition mutation was proven to be somatically acquired. Allelic loss was also noted on chromosome 18q at a marker near Smad4 in this mutated gastric cancer, apparently producing complete inactivation of Smad4 in this tumor. Significant 18q allelic loss (56% of 34 informative cases) was noted in our gastric carcinomas using microsatellite markers near the Smad4 locus, regardless of histological subtype or pathological stage. Additionally, three cases of microsatellite instability were observed. Thus, Smad4 inactivation was noted in our gastric carcinomas; however, this event was rare. The frequent loss of chromosomal arm 18q observed in gastric cancers suggests the presence of other tumor suppressor genes in this region that are involved in gastric tumorigenesis. Further studies are needed to identify these other targets of inactivation during gastric cancer development. PMID- 9331081 TI - Identification of cellular TSG101 protein in multiple human breast cancer cell lines. AB - tsg101 was identified as a tumor susceptibility gene by homozygous functional inactivation of allelic loci in mouse 3T3 fibroblasts. The human homologue was mapped at chromosome 11p15.1-2 and found to have intragenic deletion in 7 of 15 breast cancer specimens. To further confirm the relevance of defects in this gene to breast cancer, antibodies specific for the putative gene product were prepared and used to identify cellular TSG101 protein. The antibodies recognized a 46-kDa protein in human retinoblastoma WERI-27 cells labeled with [35S]methionine. This protein was not detected with preimmune sera. In cell fractionation studies, the 46-kDa protein cofractionating with glutathione S-transferase was found mainly in the cytoplasm. Similarly, when cells were immunostained with anti-TSG101 antibodies, fluorescence was localized in the cytoplasm of most of the cells. A full-size 46-kDa TSG101 protein was detected in a panel of 10 breast cancer cell lines and 2 normal breast epithelial cell lines with the same antibodies. Consistently, the full-length TSG101 mRNA was also detected in these breast cells using reverse transcription-PCR. These results indicate that homozygous intragenic deletion of TSG101 is rare in breast cancer cells. PMID- 9331082 TI - Oxidation of cyclophosphamide to 4-hydroxycyclophosphamide and deschloroethylcyclophosphamide in human liver microsomes. AB - We have investigated the formation of 4-hydroxycyclophosphamide (HCY) and deschloroethylcyclophosphamide (DCCY) from cyclophosphamide (CY) in human liver microsomes. For HCY, the estimated values (mean +/- SD; n = 3) of Km1 and Km2 were 0.095 +/- 0.072 and 5.09 +/- 4.30 mM, and the estimated values of Vmax1 and Vmax2 were 0.138 +/- 0.070 and 1.55 +/- 0.50 nmol/min/mg protein. For DCCY, Km1 and Km2 were 0.046 +/- 0.017 and 8.58 +/- 5.84 mM, and Vmax1 and Vmax2 were 0.006 +/- 0.003 and 0.274 +/- 0.214 nmol/min/mg protein. At CY concentrations of 0.1, 0.7, and 5 mM, HCY respectively accounted for 95.7 +/- 1.3, 95.1 +/- 2.4, and 90.7 +/- 2.7% of the total products of CY (HCY + DCCY; n = 6). In a separate experiment, 98.7 +/- 11.9% (n = 3) of CY loss could be accounted for by the formation of HCY at 0.1 mM CY. On the basis of cytochrome P450 (CYP) isoform specific chemical inhibitor and cDNA-expressed human P450 isozyme studies, CYP2C9 and CYP3A4/5 seemed to be the major P450 isoforms responsible for HCY formation at low (0.1 mM) and high (0.7 and 5 mM) concentrations of CY, respectively. Although orphenadrine inhibition was observed in human liver microsomes (which has been taken to indicate CYP2B6 catalysis), orphenadrine inhibited cDNA expressed CYP3A4 formation of HCY to the same extent observed in human liver microsomes, and the addition of orphenadrine to incubations containing sulfaphenazole (a specific inhibitor of CYP2C9) or troleandomycin (a specific CYP3A inhibitor) did not increase inhibition beyond that observed with sulfaphenazole or troleandomycin alone. Similar studies indicated that CYP3A4/5 was the major P450 isoform responsible for DCCY formation at high (0.7 and 5 mM) concentrations of CY. The P450 isoform responsible for DCCY formation at 0.1 mM CY could not be identified due to its very low formation rate. PMID- 9331083 TI - Nucleocytoplasmic functionality of metallothionein. AB - Appropriate nucleocytoplasmic partitioning of proteins can direct diverse cellular processes. Metallothioneins (MTs) are thiol-rich, stress-inducible proteins that can afford protection against oxidants, mutagens, and anticancer drugs. MTs display discrete nucleocytoplasmic sequestration patterns despite their small size (Mr 6,000). We demonstrate subcellular location-specific functionality of MT using a regulated expression system that restricts MT expression to the nucleus or the cytoplasm in MT-null fibroblasts. Specifically, we found that cytoplasmic but not nuclear expression of MT decreases the level of intracellular reactive oxygen species and is more cytoprotective against the prototypic oxidizing agent tert-butyl hydroperoxide. Cytoplasmic MT expression also protects against the cytotoxicity of the heavy metal CdCl2, whereas nuclear expression protects against the cytotoxicity of the mutagenic agent N-methyl-N' nitro-N-nitrosoguanidine. These data support the hypothesis that essential cytotoxic targets of both oxidants and heavy metals reside in the cytoplasm and establish the importance of nucleocytoplasmic partitioning for the function of small protective proteins such as MTs. PMID- 9331084 TI - Oxythiamine and dehydroepiandrosterone inhibit the nonoxidative synthesis of ribose and tumor cell proliferation. AB - This study investigates the significance of the glucose-6-phosphate dehydrogenase (G6PD) catalyzed oxidative and the transketolase (TK) catalyzed nonoxidative pentose cycle (PC) reactions in the tumor proliferation process by characterizing tumor growth patterns and synthesis of the RNA ribose moiety in the presence of respective inhibitors of G6PD and TK. Mass spectra analysis of 13C-labeled carbons revealed that these PC reactions contribute to over 85% of de novo ribose synthesis in RNA from [1,2-(13)C]glucose in cultured Mia pancreatic adenocarcinoma cells, with the fraction synthesized through the TK pathway predominating (85%). Five days of treatment with the TK inhibitor oxythiamine (OT) and the G6PD inhibitor dehydroepiandrosterone-sulfate (0.5 microM each) exerted a 39 and a 23% maximum inhibitory effect on cell proliferation in culture, which was increased to 60% when the two drugs were administered in combination. In vivo testing of 400 mg/kg OT or dehydroepiandrosterone-sulfate in C57BL/6 mice hosting Ehrlich's ascitic tumor cells revealed a 90.4 and a 46% decrease in the final tumor mass after 3 days of treatment. RNA ribose fractional synthesis through the TK reaction using metabolites directly from glycolysis declined by 9.1 and 23.9% after OT or the combined treatment, respectively. Nonoxidative PC reactions play a central regulating role in the carbon-recruiting process toward de novo nucleic acid ribose synthesis and cell proliferation in vitro and in vivo. Therefore, enzymes or substrates regulating the nonoxidative synthesis of ribose could also be the sites to preferentially target tumor cell proliferation by new anticancer drugs. PMID- 9331085 TI - Increased alpha2,6 sialylation of N-glycans in a transgenic mouse model of hepatocellular carcinoma. AB - Liver cancer is one of the most frequent and lethal malignancies worldwide. Early detection is hampered by the absence of reliable markers. Mice transgenic for the SV40 large T antigen under the control of a liver-specific promoter spontaneously develop well-differentiated hepatocellular carcinomas between 8 to 10 weeks of age. They are excellent models to investigate the alterations of protein expression in the early stages of tumor development and to follow these changes during tumor progression. In the present study, we analyzed the glycosylation changes occurring during tumor development in transgenic mice expressing the SV40 T antigen under the control of the antithrombin III promoter. The analysis of serum and liver glycoproteins by an ELISA type assay, using the lectin from Sambucus nigra (SNA) as a probe, revealed the presence of increased levels of Neu5Ac alpha2,6Gal beta1,4GlcNAc on N-glycans in the tumor-bearing transgenic mice as compared to controls. On serum glycoproteins the increase in alpha2,6 sialylation followed tumor progression, reaching up to 10 times control levels. However, significantly higher SNA binding (2-fold) could already be observed on serum glycoproteins from mice exhibiting only microscopically small neoplastic foci. On liver membrane glycoproteins, the increase in alpha2,6 sialylation was less pronounced, reaching two to three times control values in 6-month-old mice. Western blotting of serum and liver proteins with radiolabeled SNA showed that all glycoproteins that bind the lectin in controls exhibit larger amounts of Neu5Ac alpha2,6Gal beta1,4GlcNAc on N-glycans in the tumor-bearing mice. This general increase in alpha2,6 sialylation on all glycoproteins is due to the increased activity of the galactoside:alpha2,6 sialyltransferase (ST6Gal I), which specifically transfers Neu5Ac residues in alpha2,6 linkage to Gal beta1,4GlcNAc units on N-glycans. As for the structures synthesized by the enzyme, the increase of ST6Gal I activity in the serum as well as in liver microsomes of the transgenic mice followed tumor progression. Interestingly, the activity of the galactoside:alpha2,3 sialyltransferase (ST3Gal III), which uses the same acceptor substrate (Gal beta1,4GlcNAc), was unchanged in the earlier stages of tumor development but decreased in the serum and in liver microsomes from later stages. Using a rat ST6Gal I cDNA as a probe, Northern blots of total RNA extracted from the livers of control and transgenic mice revealed an increased (4-fold) expression of the ST6Gal I gene. The single transcripts detected in both normal and cancerous liver showed identical size. PMID- 9331086 TI - Protection conferred by selenium deficiency against aflatoxin B1 in the rat is associated with the hepatic expression of an aldo-keto reductase and a glutathione S-transferase subunit that metabolize the mycotoxin. AB - Fischer 344 rats fed on a diet that is deficient in selenium are more resistant to the hepatocarcinogen aflatoxin B1 (AFB1) than those fed on a selenium sufficient diet. Hepatic cytosol from either selenium-deficient Fischer 344 rats or Hooded Lister rats possesses a marked increase in both reductase activity toward AFB1-dialdehyde and glutathione S-transferase (GST) activity toward AFB(1) 8,9-epoxide than hepatic cytosol from selenium-sufficient rats. The elevation in hepatic AFB1-aldehyde reductase (AFAR) activity in selenium-deficient animals is accompanied by an increase of 11- and 15-fold in the levels of AFAR protein in liver cytosol from Fischer 344 and Hooded Lister rats, respectively. The amount of AFAR protein in selenium-sufficient and -deficient Fischer rats was modulated by treatment with N-acetylcysteine; this antioxidant reduced basal expression of AFAR but did not modulate the relative overexpression of AFAR during selenium deficiency. The enhanced capacity to conjugate glutathione with AFB(1)-8,9 epoxide in selenium-deficient livers from Fischer 344 and Hooded Lister rats is associated with a 5- and 7-fold increase, respectively, in the hepatic levels of the AFB1-metabolizing alpha-class GSTA5 subunit. The elevated levels of AFAR and GSTA5 protein in the selenium-deficient animals coincided with increases in the steady-state levels of their mRNAs. In selenium-deficient Fischer 344 rats, AFAR and GSTA5 were both found to be expressed throughout the centrilobular and midzonal areas of the liver lobule but were essentially absent from periportal hepatocytes. The effect of selenium insufficiency is pleiotropic, and it was also noted that the theta-class GSTT1 is overexpressed 3- and 10-fold in livers of selenium-deficient Hooded Lister and Fischer 344 rats. Inasmuch as GSTT1 is responsible for the metabolic activation of dihaloalkanes, selenium deficiency may increase the susceptibility of rats to mutagens such as dichloromethane. PMID- 9331087 TI - Sulindac causes rapid regression of preexisting tumors in Min/+ mice independent of prostaglandin biosynthesis. AB - Several lines of evidence strongly link prostaglandins (PGs) and leukotrienes (LTs) to cancer of the intestine. Several studies have reported a 40-50% reduction in mortality from colorectal cancer in individuals who routinely consume nonsteroidal anti-inflammatory drugs, possibly by inhibiting cyclooxygenase activity. However, the role of eicosanoids in this process is still unclear. The heterozygote Min/+ mouse model, like patients with familial adenomatous polyposis, carries a nonsense mutation in the adenomatous polyposis coli (APC) gene that results in the spontaneous development of intestinal adenomas (100% incidence). This study investigated the association between eicosanoid biosynthesis, intestinal tumor load, and the chemotherapeutic effect of the nonsteroidal anti-inflammatory drug sulindac during early and preexisting phases of tumor growth and development as well as residual effects after drug withdrawal. Administration of sulindac (320 ppm) to Min/+ mice reduced the tumor number by 95% but did not alter the levels of PGE2 and LTB4 in intestinal tissues. Increasing PGE2 and LTB4 levels by 44% with dietary arachidonic acid supplementation had no effect on tumor number or size. When sulindac was added to the arachidonic acid-supplemented diet, tumor number was reduced by 82%, whereas eicosanoid levels remained elevated. In Min/+ mice with established tumors, treatment with sulindac for 4 days reduced tumor number by 75%, and continual administration of sulindac was necessary to maintain a reduced tumor load. In summary, alterations in eicosanoid formation were not correlated with tumor number or size in the Min/+ mouse model; thus, the antitumor effect of sulindac seems to be PG independent. PMID- 9331088 TI - Earlier onset of melanotroph carcinogenesis in mice with inherited mutant paternal allele of the retinoblastoma gene. AB - The role of genomic imprinting in the development of tumors with defective retinoblastoma protein function remains debatable. Disruption of either parental allele of the murine retinoblastoma (Rb) gene is sufficient for spontaneous melanotroph carcinogenesis to occur in almost all Rb+/- mice. Nevertheless, mice with a disrupted paternal Rb allele succumb to tumors faster. In these animals, the first foci of proliferating atypical Rb-negative cells appear and progress to overtly malignant tumors earlier. In addition, more foci of early atypical proliferation are observed. In Rb+/- mice, however, parental origin influences neither Rb expression nor proliferation of melanotrophs. Accordingly, Rb-/- mice rescued by the human RB transgene transmitted either paternally or maternally have similar survival rates. Taken together, the data point to the existence of an imprinted gene in an Rb-linked locus. The function of this gene affects the onset of melanotroph carcinogenesis, likely by controlling preferential survival of the cells with secondary loss of the Rb maternal allele. Rb+/- mice may serve as useful models to identify and characterize genomic imprinting mechanisms influencing carcinogenesis associated with Rb loss of function. PMID- 9331089 TI - In vivo selective gene expression and therapy mediated by adenoviral vectors for human carcinoembryonic antigen-producing gastric carcinoma. AB - Previously, we reported that adenoviral vectors carrying the carcinoembryonic antigen (CEA) promoter sequences to direct the Echerichia coli beta-galactosidase gene (AdCEA-lacZ) or cytosine deaminase (CD) gene (AdCEA-CD) confer selective gene expression on a CEA-positive gastric cancer cell line (MKN45) in vitro. Here, adenovirus-mediated tumor-specific gene therapy for CEA-positive gastric carcinoma in vivo was investigated. Using an animal model with i.p. disseminated MKN45 tumors, adenovirus-mediated tumor-specific transgene expression and therapeutic efficacy were analyzed. After an i.p. injection of AdCEA-lacZ, beta galactosidase activity was confined to tumor xenografts. Moreover, CD mRNA was expressed exclusively in MKN45 tumor xenografts after infection with AdCEA-CD, despite the fact that an adenovirus-mediated transfer of CD DNA was detected in all tissues tested. In contrast, CD mRNA was detected not only in tumor xenografts but also in other organs of mice infected with AdCA-CD, in which CD gene expression is governed by an ubiquitous promoter. Suppression of tumor growth and prolongation of survival were noted in tumor-bearing mice treated with AdCEA-CD and 5-fluorocytosine (5FC) without observable adverse effects. In contrast, significant hepatic toxicity was noted in animals treated with AdCA-CD. These results reveal that the CEA promoter restricts CD gene expression to CEA positive tumor cells in the adenoviral context in vivo, along with the beneficial therapeutic effects of 5FC treatment, suggesting the i.p. AdCEA-CD/5FC system may provide a novel approach to treatment of i.p. disseminated gastric cancer. PMID- 9331090 TI - Characterization of the p53 tumor suppressor pathway in cell lines of the National Cancer Institute anticancer drug screen and correlations with the growth inhibitory potency of 123 anticancer agents. AB - In the present study, we report the characterization of the p53 tumor suppressor pathway in the 60 cell lines of the National Cancer Institute (NCI) anticancer drug screen, as well as correlations between the integrity of this pathway and the growth-inhibitory potency of 123 anticancer agents in this screen. Assessment of p53 status in these lines was achieved through complete p53 cDNA sequencing, measurement of basal p53 protein levels and functional assessment of (a) transcriptional activity of p53 cDNA from each line in a yeast assay, (b) gamma ray-induced G1 phase cell cycle arrest, and (c) gamma-ray-induced expression of CIP1/WAF1, GADD45, and MDM2 mRNA. Our investigations revealed that p53 gene mutations were common in the NCI cell screen lines: 39 of 58 cell lines analyzed contained a mutant p53 sequence. cDNA derived from almost all of the mutant p53 cell lines failed to transcriptionally activate a reporter gene in yeast, and the majority of mutant p53 lines studied expressed elevated basal levels of the mutant p53 protein. In contrast to most of the wild-type p53-containing lines, cells containing mutant p53 sequence were also deficient in gamma-ray induction of CIP1/WAF1, GADD45, and MDM2 mRNA and the ability to arrest in G1 following gamma-irradiation. Taken together, these assessments provided indications of the integrity of the p53 pathway in the 60 cell lines of the NCI cell screen. These individual p53 assessments were subsequently used to probe a database of growth inhibitory potency for 123 "standard agents," which included the majority of clinically approved anticancer drugs. These 123 agents have been tested against these lines on multiple occasions, and a proposed mechanism of drug action had previously been assigned to each agent. Our analysis revealed that cells with mutant p53 sequence tended to exhibit less growth inhibition in this screen than the wild-type p53 cell lines when treated with the majority of clinically used anticancer agents: including DNA cross-linking agents, antimetabolites, and topoisomerase I and II inhibitors. Similar correlations were uncovered when we probed this database using most of the other indices of p53 status we assessed in the lines. Interestingly, a class of agents that differed in this respect was the antimitotic agents. Growth-inhibitory activity of these agents tended, in this assay, to be independent of p53 status. Our characterization of the p53 pathway in the NCI cell screen lines should prove useful to researchers investigating fundamental aspects of p53 biology and pharmacology. This information also allows for the large-scale analysis of the more than 60,000 compounds tested against these lines for novel agents that might exploit defective p53 function as a means of preferential toxicity. PMID- 9331091 TI - Role of topoisomerase II beta in the resistance of 9-OH-ellipticine-resistant Chinese hamster fibroblasts to topoisomerase II inhibitors. AB - In the Chinese hamster lung cell line DC-3F/9-OH-E, made resistant to 9-OH ellipticine and cross-resistant to other topoisomerase II inhibitors, the amount of topoisomerase II alpha is 4-5-fold lower than in the parental DC-3F cells. A mutation in position 1710 of topoisomerase II beta cDNA, generating a stop codon, completely abolishes the expression of this isoform in DC-3F/9-OH-E cells. To analyze the contribution of the loss of topoisomerase II beta to the resistance phenotype, DC-3F/9-OH-E cells were cotransfected with two plasmids, one conferring the resistance to G418, the other carrying the topoisomerase II beta cDNA. Among 200 G418-resistant clones, one was found to contain a topoisomerase II beta activity similar to that in the parental cells. These cells constitute an in vivo mammalian model to study the pharmacological role of topoisomerase II beta. In the transfected cells, different levels of cleavable complex formation and resistance reversion were observed with each topoisomerase II inhibitor examined. This work demonstrates that topoisomerase II beta is a pharmacological target for 9-OH-ellipticine, etoposide, or 4'-(9-acridinylamino)methanesulfon-m anisidide and plays a role in the cytotoxicity of these agents. Furthermore, topoisomerase II beta is the preferential target for 4'-(9 acridinylamino)methanesulfon-m-anisidide. The loss of topoisomerase II beta activity in the DC-3F/9-OH-E cells is then in part responsible for their resistance to topoisomerase II inhibitors. PMID- 9331092 TI - Growth inhibitory effects of sodium phenylacetate (NSC 3039) on ovarian carcinoma cells in vitro. AB - The aim of this study was to determine the antiproliferative activity of sodium phenylacetate (NaPa) against ovarian carcinoma cell lines. NaPa induced a dose dependent inhibition (IC50 from 12 mM to >20 mM) of all ovarian carcinoma cell lines, although the sensitivity of individual lines to NaPa varied. Both cisplatin-sensitive and -resistant cell lines responded to NaPa, and growth inhibitory activity was also detected against cells freshly isolated from malignant ascites of previously treated patients. The growth inhibitory effects that were produced by NaPa were time dependent, showing a maximum effect at 72 h, and were not associated with cytotoxic action. Growth inhibitory effects of NaPa were also reversible. After 48- and 72-h exposures to NaPa, a reduction in the percentage of cells in the S-phase was detected, with a concomitant recruitment of cells in the G0-G1 phase. Treatment with NaPa after different exposure times did not significantly increase the proportion of cells undergoing apoptosis. NaPa also produced a significant reduction in the percentage of cyclin-D1- and p21/ras positive cells and in the percentage of cells positive for bcl-2, whereas the percentages of bax/p21-positive cells increased. NaPa produced minimal, if any, alterations of expression of HLA class I and transforming growth factor beta1 antigens. In contrast, the percentage of transforming growth factor beta2 positive cells decreased after exposure to NaPa. The combination of NaPa with cisplatin resulted in an additive inhibitory effect. Our results show, for the first time, that NaPa inhibits the growth of ovarian carcinoma cell lines and the cells from malignant ascites of chemotherapy-treated patients with ovarian carcinoma. The growth-inhibitory properties of NaPa suggest that this molecule could represent a prototype of a new class of compounds with possible therapeutic potential in patients with ovarian carcinoma. PMID- 9331093 TI - R-flurbiprofen chemoprevention and treatment of intestinal adenomas in the APC(Min)/+ mouse model: implications for prophylaxis and treatment of colon cancer. AB - We used the C57BL/6J-APC(Min)/+ mouse (Min mouse) to evaluate the chemopreventive effects of R-flurbiprofen (R-FB), the noncyclooxygenase-inhibiting enantiomer of FB. Weanling Min mice were administered 6 weeks of oral treatment with R-FB using 2.5-25 mg/kg of R-FB once per day (q.d.), 2.5-10 mg/kg of R-FB twice per day (b.i.d.), and 5 mg/kg of R-FB b.i.d. challenged with a high saturated fat diet. At necropsy we determined tumor and ulcer numbers, tumor size, and plasma levels of R- and S-FB. A linear dose response was observed from 2.5 to 10 mg/kg of R-FB, regardless of whether the drug was administered as a single or divided dose. Reductions in tumor number were significant (P < or = 0.02) for doses of R-FB from 2.5 to 25 mg/kg/day. A dose of 5 mg/kg R-FB b.i.d. was able to overcome the doubling in tumor number associated with the high saturated fat diet. At 20 and 25 mg/kg/day R-FB, we obtained the maximum response with up to 90% inhibition of total tumor number. At these doses, however, there was toxicity and animal deaths. This toxicity was associated with ulceration, presumably resulting from the in vivo epimerization of R- to S-FB that occurs in the mouse. Thus, we evaluated the oral pharmacokinetics of R-FB and its conversion to S-FB in wild type mice. These kinetics experiments revealed inversion rates of 7.3 and 11.0% for the 2.5 and 10 mg/kg R-FB doses, respectively. S-FB administered alone (0.5 and 2.0 mg/kg q.d.), in doses mimicking the concentrations of S-FB associated with the R to S epimerization of the doses of R-FB used in our experiments, had little or no antitumor efficacy (P > 0.05). Thus, we conclude that R-FB itself, not the S-FB resulting from epimerization in the mouse, inhibits adenoma formation in the Min mouse. In humans, where there is no R to S epimerization, it is possible that larger doses of R-FB can be used without causing cyclooxygenase inhibition and its resulting ulcerogenicity and other side effects. To assess the effect of R-FB on established adenomas, we allowed 40 Min mice to remain untreated until 70 days of age (the time of necropsy in the previous experiments) and then treated them for an additional 42 days with 10 mg/kg R-FB q.d. or 5 mg/kg R-FB b.i.d.. Both drug-treated groups demonstrated tumor numbers significantly less than that of the vehicle control (P < 0.01). Our results suggest that prophylaxis and treatment trials of R-FB should be extended to humans. PMID- 9331094 TI - Molecular chemotherapy combined with radiation therapy enhances killing of cholangiocarcinoma cells in vitro and in vivo. AB - Cholangiocarcinoma is a virtually incurable tumor, resistant to current surgical, chemotherapy, and radiotherapy interventions. We applied the gene therapy strategy of toxin gene conversion of nontoxic prodrug to chemotherapeutic drug in combination with radiation therapy to the treatment of cholangiocarcinoma. In this regard, 5-fluorouracil (5-FU) is an accepted radiosensitizing and chemotherapeutic agent presently used in cancer therapy. The Escherichia coli enzyme cytosine deaminase (CD) converts the prodrug 5-fluorocytosine (5-FC) to 5 FU. Therefore, our goal was to express the CD gene in the human cholangiocarcinoma cell line, SK-ChA-1, assess the cytotoxicity of intracellular production of 5-FU, and determine any enhanced cell killing by the addition of external beam radiation. The susceptibility of SK-ChA-1 cells to recombinant adenoviral infection was determined by fluorescence-activated cell sorting analysis. We used the recombinant adenoviral vector AdCMVLacZ, encoding the E. coli beta-galactosidase reporter gene under control of the human cytomegalovirus (CMV) promoter, to infect SK-ChA-1 and HeLa cells at 10 and 100 plaque forming units (pfu)/cell, followed by FACS analysis. To evaluate CD-mediated conversion of 5-FC to 5-FU and subsequent cytotoxicity, SK-ChA-1 cells were infected with the recombinant adenovirus AdCMVCD, which encodes CD. Cells were then plated in 96-well microtiter plates and exposed to varying concentrations of 5-FC. Cell proliferation assays (tetrazolium salt conversion to formazan colorimetric assay) were performed beginning 2-8 days after plating. We evaluated the effects of external beam radiation using a single 8 Gy 60Co dose to AdCMVCD infected cells, with prior exposure to 5-FC for 2-3 days. MTS assays were performed following radiation treatment. Radiation dose-response analysis, via clonogenic assay, was used as a more sensitive assay to confirm the interaction of the treatment conditions. s.c. SK-ChA-1 tumors in athymic nude mice were established, which then received three intratumoral injections of 1 x 10(9) pfu AdCMVCD. Mice received i.p. injections of 400 mg/kg of 5-FC twice daily for 7 days beginning the day of initial AdCMVCD injection (day -2). The radiation treatment group received 10 Gy of 60Co exposure to their tumor on day 0. SK-ChA-1 cells were efficiently transduced (48.7 and 99.2%) by 10 and 100 pfu/cell of AdCMVLacZ, respectively. From 37.9 to 84.4% of SK-ChA-1 cells were killed following infection with 10 pfu/cell AdCMVCD and 8 days of exposure to various concentrations of 5-FC (5, 10, 30, 50, and 100 microg/ml). Higher 5-FC concentrations and longer duration of exposure resulted in greater cell killing. Radiation treatment (8 Gy) enhanced cell killing by greater than 70% when combined with 10 or 20 microg/ml of 5-FC. Radiation dose-response analysis with clonogenic assay confirmed enhanced SK-ChA-1 cell cytotoxicity as a result of radiation treatment following AdCMVCD infection and 5-FC exposure, with radiobiological parameters alpha = 0.44 and D0 = 0.96. Combined treatment of SK ChA-1 tumors with AdCMVCD, 5-FC, and radiation in animals resulted in significantly greater survival, time to tumor regrowth, and doubling time compared to the nonradiation treatment group (P = 0.03, 0.015, and 0.002, respectively). Significantly greater change in tumor size, smaller ratio of final tumor size to original tumor size, and smaller final tumor size were observed in the radiation treatment group compared to the no radiation treatment group (P = 0.02, 0.03, and 0.03, respectively). Human cholangiocarcinoma cells were transduced with a recombinant adenovirus in vitro at high efficiency and were susceptible to CD-mediated intracellular 5-FU production. Radiobiological survival curve parameters confirmed an interactive cytotoxic effect when viral infection and prodrug therapy were combined with external beam radiation exposure. (ABSTRACT TRUNCATED) PMID- 9331095 TI - Intratumoral delivery of boronated epidermal growth factor for neutron capture therapy of brain tumors. AB - The gene for epidermal growth factor receptor (EGFR) is amplified or overexpressed in high-grade gliomas but is low or undetectable in normal brain. Recently, there has been increasing interest in using epidermal growth factor (EGF)-based bioconjugates as targeting agents for brain tumors. In the present study, we have investigated the potential use of boronated EGF as a delivery agent for boron neutron capture therapy, which is based on the capture reaction that occurs when 10B, a stable isotope, is irradiated with low-energy thermal neutrons. A fourth generation starburst dendrimer was boronated and linked to EGF using heterobifunctional reagents. Either wild-type or EGFR gene transduced C6 glioma cells (C6EGFR), which expressed 10(5)-10(6) receptor sites/cell, were stereotactically implanted into the right cerebral hemisphere of Fischer rats. Four weeks later, the rats received either i.v. or intratumoral (i.t.) injection of 131I-labeled boronated starburst dendrimer (BSD) or BSD-EGF. The biodistribution of 131I-BSD-EGF and 131I-BSD was studied by means of whole-body scintigraphy, autoradiography, and gamma scintillation counting. Following i.t. injection of 131I-BSD-EGF, 21.8% of the injected dose per gram tissue (% ID/g) was localized in C6EGFR tumors at 24 h and 16.3% at 48 h compared to 5 and 1.3% ID/g in C6 wild-type tumors, respectively, and 0.01 and 0.006% ID/g, respectively, for i.v. injected animals at the corresponding times. In contrast, following i.t. injection of BSD-EGF, only 0.01-0.1% ID/g was localized in the liver and spleen at 24 and 48 h compared to 5-12% ID/g following i.v. injection. Our data indicate that direct i.t. injection can selectively deliver BSD-EGF to EGFR-positive gliomas and suggest that intracerebral administration may be the most effective way for delivering EGF-based bioconjugates to EGFR-positive brain tumors. PMID- 9331096 TI - Decreasing the apoptotic threshold of tumor cells through protein kinase C inhibition and sphingomyelinase activation increases tumor killing by ionizing radiation. AB - Approximately 30% of cancer deaths result from the failure to control local and regional tumors. The goal of radiotherapy is to maximize local and regional tumor cell killing while minimizing normal tissue destruction. Attempts to enhance radiation-mediated tumor cell killing using halogenated pyrimidines, antimetabolites, and other DNA-damaging agents or sensitizers of hypoxic tumor cells have met with only modest clinical success. In an unique strategy to modify tumor radiosensitivity, we used an inhibitor of the protein kinase C group A and B isoforms, chelerythrine chloride (chelerythrine), to enhance the killing effects of ionizing radiation (IR). Protein kinase C activity plays a central role in cellular proliferation, differentiation, and apoptosis. Chelerythrine increases sphingomyelinase activity and enhances IR-mediated cell killing through induction of apoptotic tumor cell death in a radioresistant tumor model both in vitro and in vivo. Although previous reports have suggested that IR-mediated apoptosis correlates with tumor volume reduction, we demonstrate for the first time that lowering the apoptotic threshold increases tumor cell killing in vivo. PMID- 9331097 TI - Synthetic peptides derived from the melanocyte-stimulating hormone receptor MC1R can stimulate HLA-A2-restricted cytotoxic T lymphocytes that recognize naturally processed peptides on human melanoma cells. AB - Human melanoma-specific HLA-A2 restricted CTLs have recently been shown to recognize antigens expressed by melanoma lines and normal melanocytes, including Melan-A/Mart-1, gp100, gp75, and tyrosinase. Herein, we define HLA-A2-restricted CTL epitopes from a recently cloned melanocortin 1 receptor (MC1R), which belongs to a new subfamily of the G-protein-coupled receptors expressed on melanomas and melanocytes. Thirty-one MC1R-derived peptides were selected on the basis of HLA A2-specific motifs and tested for their HLA-A2 binding capacity. Of a group of 12 high or intermediate HLA-A2 binding peptides, three nonamers, MC1R244 (TILLGIFFL), MC1R283 (FLALIICNA), and MC1R291 (AIIDPLIYA), were found to induce peptide-specific CTLs from peripheral blood mononuclear cells of healthy HLA-A2+ donors after repeated in vitro stimulation with peptide-pulsed antigen-presenting cells. The CTLs raised against these three HLA-A2+-restricted peptides could recognize naturally processed peptides from HLA-A2+ melanomas and from Cos7 cells cotransfected with MC1R and HLA-A2. CTLs induced by the MC1R291 peptide (but not induced or induced only to a very low extent by the other two MCR1 peptide epitopes) showed cross-reactions with two other members of the melanocortin receptor family, which are more broadly expressed on other tissues. Taken together, our findings have implications in relation both to autoimmunity and immunotherapy of malignant melanomas. PMID- 9331098 TI - Developmental exposure to diethylstilbestrol elicits demethylation of estrogen responsive lactoferrin gene in mouse uterus. AB - Alteration of DNA demethylation in five CpG sites (-547, -533, -475, -464, and 454) immediately upstream from the estrogen response element of lactoferrin promoter was determined in the uteri of immature (17-day-old) and mature (21- and 30-day-old) mice treated neonatally with DES. Only the CpG/-464 was found to be abnormally demethylated by diethylstilbestrol (DES) treatment in the mature uteri. This abnormal demethylation occurred in specific response to DES in neonatal mice, because DES injected into the 30-day-old mature mice did not demethylate CpG/-464. This site, however, remained methylated in the neonatally DES-treated/ovariectomized mice, indicating that this DES-elicited demethylation is under hormonal control. Thus, neonatal DES treatment appeared to imprint an abnormal, site-specific demethylation of CpG/-464, which requires ovarian hormones to occur in adult mice. Moreover, the demethylation was maintained in uterine tumors of the neonatally DES-treated mice. This mode of demethylation is reminiscent of uterine tumor formation, which also depends on both neonatal DES exposure and ovarian hormone stimulation in adulthood. Thus, neonatal DES treatment may induce tumor formation as well as demethylation through a common cellular process. PMID- 9331100 TI - Comparative genomic hybridization analysis of breast tumors with predetermined profiles of DNA amplification. AB - In a separate study (F. Courjal et al., Cancer Res., 57: 4360-4367, 1997), we have analyzed by Southern blotting the relationship between DNA amplification and clinicopathological features of breast cancer. Six regions of recurrent amplifications were tested (8p12, 8q24, 11q13, 12q13, 17q12, and 20q13), and the results suggested that there was a relationship between DNA amplification profiles and breast tumor phenotype. We had delineated three subgroups of tumors showing distinct DNA amplification profiles and clinicopathological characteristics: group A, tumors showing amplification at 11q13 and/or 8p12 and/or 20q13; group B, tumors amplified at ERBB2 and/or MYC and/or MDM2/SAS; and group C, tumors with no detectable amplification. The aim of the present work was to characterize extensively the amplification profiles in the different subgroups of tumors. Sixty-one breast tumors distributed in all three subgroups were studied by comparative genomic hybridization (CGH). There was an overall good agreement between Southern blotting results and CGH data. As expected, CGH revealed gains undetected by Southern blotting. Most of these gains occurred in regions for which no adapted probes were available but also revealed nondetected amplifications at 8q24 or 20q13. Tumors showed multiple aberrations with a medium number of 5.6 copy number variations/tumor, whereas, according to Southern blotting results, 38% of the tumors analyzed were devoid of any amplification. This proportion fell to 6.5% after CGH analysis. Recurrent gains were observed in tumors from all three subgroups, albeit at varying incidences, and involved 1q, 8q, 17q23-q24, and 20q13. Gains covered large regions of DNA and could possibly include several cores of amplification. Some events, such as gains at 16p11-p12 and 14q or losses at 22q, showed more restricted distributions, suggesting the existence of additional sets of preferential coamplifications. The complexity of genetic profiles revealed by CGH indicates that breast cancer development depends on a large (yet undetermined) number of genetic events. The description of molecular phenotypes in breast cancer may therefore prove to be complex, and it should be interesting to see how many breast tumor subtypes will be defined in the end. PMID- 9331099 TI - Mapping of DNA amplifications at 15 chromosomal localizations in 1875 breast tumors: definition of phenotypic groups. AB - DNA amplification is frequent in breast cancer and has been associated with specific clinicopathological parameters and/or worsened course of the disease. In the present work, we were interested in further defining the association linking the occurrence of DNA amplification to the emergence of specific breast tumor phenotype. To this aim, we studied by Southern blotting a total of 1875 breast tumor DNAs with 26 probes mapping at 15 distinct chromosomal localizations. Of the 26 loci tested, 11 loci showed elevated levels of amplification, 9 loci showed occasional and/or low level of DNA copy number increase, and 6 loci showed very rare or no variation. This allowed us to define six amplified domains mapping at 8p12, 8q24, 11q13, 12q13, 17q12, and 20q13.2, respectively. Over 60% of the tumors analyzed presented at least one amplification at one of these localizations. Amplifications often covered large regions of DNA and bore complex patterns involving coamplification of several colocalized markers. Statistical analysis revealed correlations associating DNA amplification with breast tumor phenotype, as well as sets of preferential coamplifications. Based on these correlations, we defined three subsets of breast cancer according to their patterns of DNA amplification. The first subset (group A) was organized around the amplifications at 11q13 and/or 8p12 and was predominantly composed of estrogen receptor-positive tumors and presented a large proportion of lobular cancers. The second subset (group B) was organized around the amplifications of ERBB2 and/or MYC. These tumors were mostly estrogen receptor-negative and of the ductal invasive type. The third subset (group C) corresponded to tumors in which no amplification was detected in the present screen. Tumors in this group were largely diploid and of low histopathological grading. PMID- 9331101 TI - Polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin induce intrachromosomal recombination in vitro and in vivo. AB - Polychlorinated aromatic hydrocarbons such as polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are extremely stable and widely distributed environmental pollutants. These chemicals are animal carcinogens and probable human carcinogens, and TCDD is possibly one of the most potent toxins ever evaluated by the United States Environmental Protection Agency. Polychlorinated aromatic hydrocarbons score negatively in most genotoxicity assays, including the Ames (Salmonella) assay. Although their mechanism of toxicity is not well understood, they induce aryl hydrocarbon (AH) hydroxylases and bind to the AH receptor, which is believed to mediate toxicity. Here, we determine effects of polychlorinated aromatic hydrocarbons in genotoxicity assays that score for DNA deletions by intrachromosomal recombination in vivo and in vitro. In this study, TCDD, Aroclor 1221, and Aroclor 1260 induced deletions in vivo in the mouse embryo; Aroclor 1221 and Aroclor 1260 induced deletions in yeast. We also show that the induced deletion events did not correlate with induction of AH hydroxylase. None of the tested compounds induced CYP1A associated ethoxyresorufin-O-deethylase activity in mouse embryos or in vitro. These results clearly demonstrate a genotoxic activity of polychlorinated aromatic hydrocarbons in vitro and in vivo, which is independent of induction of cytochrome P450 activity. Because genetic instability and deletions may be mechanistically involved in carcinogenesis, these results may encourage further research to determine whether such genotoxic mechanisms may be useful for cancer risk assessment of polychlorinated aromatic hydrocarbons. PMID- 9331102 TI - A lack of radiation-induced ornithine decarboxylase activity prevents enhanced reactivation of herpes simplex virus and is linked to non-cancer proneness in xeroderma pigmentosum patients. AB - Patients with xeroderma pigmentosum (XP), a DNA repair disorder, run a large risk of developing skin cancer in sun-exposed areas. Cancer proneness in these patients correlates with a mammalian SOS-like response, "enhanced reactivation (ER) of viruses." Here, we report that radiation-induced activation of the ornithine decarboxylase (ODC) gene, a putative proto-oncogene, is required for this response. Various diploid fibroblast strains derived from a non-cancer-prone subclass of XP patients, which lack the ER response, were irradiated with 2 J/m2 and assessed for gene induction. In these fibroblasts, an absence of induction of ODC by UV-C was observed at the levels of mRNA, protein, and enzyme activity. This lack of induction is quite specific because the genes for fos and collagenase were induced as they were in normal XP cells. The apparent linkage between non-cancer proneness and a lack of ER and ODC induction was confirmed in a fibroblast strain derived from a patient with another DNA repair disorder, trichothiodystrophy, which does not lead to cancer proneness: in these cells, no induction of the ER response nor of ODC occurs after UV-C irradiation. Repair deficiency, however, is not essential because the simultaneous lack of ODC and ER induction after 10 J/m2 UV-C was found in at least one repair-proficient fibroblast. Next, a specific inhibitor of ODC, difluoromethylornithine, at a dose of 10 mM, completely blocked the ER response in cultured normal skin fibroblasts, suggesting that the ODC enzyme is in fact essential for the ER response. Difluoromethylornithine, although it did not affect other processes such as DNA repair, leads to a block in the cell division cycle at the G1-S transition. Interestingly, other blockers of this transition, wortmannin (500 nM) and mimosine (100 mM), also decreased the ER response. Finally, the ER and ODC responses also seem to be linked after treatment with X-irradiation (3 Gy), suggesting that both are part of a general response to DNA damage, at least in human skin fibroblasts. Apart from the abnormal ER and ODC responses, fibroblasts from non-tumor-prone XP patients react in the same way to radiation as do fibroblasts from tumor-prone XP patients with respect to other parameters. Thus, the lack of ODC induction after radiation may help to protect XP patients against skin carcinogenesis. PMID- 9331103 TI - Mutation frequency and spectrum in lymphocytes of small cell lung cancer patients receiving etoposide chemotherapy. AB - Etoposide, a topoisomerase II inhibitor, is a chemotherapeutic agent that is used in the treatment of a wide variety of neoplasms, including small cell lung cancer, germ cell cancer, testicular cancer, acute leukemia, and lymphoma. Although it has proven valuable, etoposide is also a known mutagen and has been implicated as a causative agent of treatment-related secondary acute nonlymphocytic leukemia. We have investigated the induction of mutation following etoposide treatment in vivo using the hypoxanthine phosphoribosyltransferase (hprt) T-cell cloning assay in small cell lung cancer patients receiving single drug etoposide chemotherapy. This report presents results on the monitoring of 12 patients (mean age, 74.8 +/- 6.0 years; range, 66-83 years) before, during, and after chemotherapy. The treatment regimen included up to six cycles of oral etoposide given in twice-daily 50-mg tablets for 10-14 days, separated by 2 weeks of rest. Peripheral blood samples were collected on the first day of each cycle prior to treatment. Patients received one to six etoposide cycles and were followed for 0.7-5.3 months after the start of chemotherapy (total etoposide dose, 1.4-8.4 g). Results from the pooled data show no significant increase in the hprt mutant frequency (pretreatment, 46 x 10(-6) +/- 38 x 10(-6), versus posttreatment, 55 x 10(-6) +/- 46 x 10(-6)), although considerable interpatient variability was observed. Of a total of 424 selected mutants, 228 were analyzed by sequencing hprt cDNA. Spectra of 56 pretreatment and 147 posttreatment mutations revealed significant enhancement of AT-->TA transversions and a concomitant decrease in the number of GC-->TA transversions in posttreatment spectra, when they were compared with pretreatment or control spectra. No evidence for the induction of gross deletions or rearrangements was found in the spectra of mutants that were recovered from patients after etoposide treatment. The lack of enhanced mutant frequency after treatment suggests that the etoposide chemotherapy was not particularly effective in inducing mutation, as measured by the hprt assay. It is proposed that mutated cells are eliminated through apoptosis due to accumulated DNA damage. PMID- 9331104 TI - The absolute number of trans-rearrangements between the TCRG and TCRB loci is predictive of lymphoma risk: a severe combined immune deficiency (SCID) murine model. AB - Pilot studies in human populations have demonstrated a correlation between the level of antigen receptor trans-rearrangements and risk (at the population level) of lymphoid malignancy. Irradiation of newborn severe combined immune deficiency mice results in an increased risk of subsequent development of thymic lymphoma (100% of mice so irradiated are dead of thymic lymphoma by 20 weeks of age). We, therefore, assayed the occurrence of trans-rearrangements in this well-controlled mouse mutant system and found a 50-100-fold increase in the absolute number of TCRGV-TCRBJ trans-rearrangements compared to unirradiated littermates (and a comparable fold increase over age-matched BALB/c mice) at 2 weeks following irradiation. We also found a marked disproportion in generating trans rearrangements versus intralocus rearrangements in the severe combined immune deficiency system compared to BALB/c, independent of irradiation. The trans rearrangements noted were polyclonal in nature. These data, again, suggest that the absolute level of antigen receptor trans-rearrangements may serve as a biomarker of lymphoma risk. PMID- 9331106 TI - Frameshift somatic mutations in gastrointestinal cancer of the microsatellite mutator phenotype. AB - An exacerbated genomic instability characterizes hereditary and sporadic gastrointestinal cancer of the microsatellite mutator phenotype (MMP), generating somatic frameshift mutations in genes containing mononucleotide repeats. We have recently shown that approximately 50, 40, and 30% of MMP+ colon tumors harbor frameshift mutations in (G)8, (A)8, and (C)8 tracks within the proapoptotic gene BAX and the hMSH3 and hMSH6 DNA mismatch repair genes, respectively. Here we report a higher incidence of frameshift mutations in these 3 genes in a panel of 25 MMP+ gastric adenocarcinomas: 64% in BAX and hMSH3, and 52% in hMSH6. These results support a multiple mutator gene model for the stepwise unfolding of the MMP and further illustrate the importance of the escape from apoptosis in gastrointestinal cancer. The tumor suppressor role played by BAX is also supported by the finding of other somatic BAX mutations, including recurrent missense mutations, not only in gastrointestinal cancer of the MMP but also in gastrointestinal cancer without the MMP. PMID- 9331105 TI - Inhibition of tumor promoter-induced activator protein 1 activation and cell transformation by tea polyphenols, (-)-epigallocatechin gallate, and theaflavins. AB - (-)-Epigallocatechin gallate (EGCG) and theaflavins are believed to be key active components in tea for the chemoprevention against cancer. However, the molecular mechanisms by which EGCG and theaflavins block carcinogenesis are not clear. We have used the JB6 mouse epidermal cell line, a system that has been used extensively as an in vitro model for tumor promotion studies, to examine the anti tumor promotion effects of EGCG and theaflavins at the molecular level. EGCG and theaflavins inhibited epidermal growth factor- or 12-O-tetradecanoylphorbol-13 acetate-induced cell transformation in a dose-dependent manner. At the dose range (5-20 microM) that inhibited cell transformation, EGCG and theaflavins also inhibited AP-1-dependent transcriptional activity and DNA binding activity. The inhibition of AP-1 activation occurs through the inhibition of a c-Jun NH2 terminal kinase-dependent, but not an extracellular signal-regulated protein kinase (Erk) 1-dependent or Erk2-dependent, pathway. Because the transcription factor AP-1 is important for tumor promoter-induced neoplastic transformation, the inhibitory effects on AP-1 activation by EGCG and theaflavins may further explain the anti-tumor promotion action of these tea constituents. PMID- 9331107 TI - Immunolocalization and messenger RNA expression of bone morphogenetic protein-6 in human benign and malignant prostatic tissue. AB - Skeletal metastases are common in advanced prostate cancer, causing considerable morbidity, and they are usually osteoblastic in nature with no clear explanation for this phenomenon. Bone morphogenetic proteins (BMPs) induce bone formation in vivo, and preliminary work showed a possible association between BMPs and prostatic skeletal metastases; differential expression favors BMP-6 as a potential new marker and mediator of osteosclerotic deposit formation. We investigated BMP-6 mRNA and protein expression by in situ hybridization and immunohistochemistry in malignant and benign prostates from 40 men. BMP-6 mRNA expression was detected exclusively in malignant epithelial cells in 20 of 21 patients (95%) with metastases and in 2 of 11 patients (18%) with localized cancer, and it was absent in 8 benign samples. Immunostaining for BMP-6 was predominantly cytoplasmic and was present in all primary tumors with established metastases and in 4 of 11 (36%) organ-confined cancers. In benign prostatic hyperplasia, basal cells and areas of basal cell hyperplasia were positive for BMP-6 by immunohistochemistry. The results suggest a close association between BMP-6 expression in primary malignant prostatic tissue and skeletal metastases. BMP-6 may be responsible, in part, for the osteoblastic changes in metastatic lesions secondary to prostate cancer. PMID- 9331108 TI - In vivo induction of murine cytokine production by carcinoembryonic antigen. AB - Carcinoembryonic antigen (CEA) may promote experimental metastasis through production of cytokines. The effect of systemic CEA on the production of proinflammatory cytokines was investigated in mice and compared to levels induced by lipopolysaccharide (LPS). Serum concentrations of interleukin (IL)-6 peaked 1 h after an i.v. CEA injection of 40 microg/mouse to 37-54% of the maximal level induced by a 1 microg/mouse injection of LPS in both normal and immunoincompetent mice. The CEA induction of IL-6 was a specific response, because the peptide PELPK (the pentapeptide on CEA that is the ligand for the CEA receptor on Kupffer cells) conjugated to albumin induced 30% of the maximal CEA response for IL-6, whereas the specificity control PELGK-conjugated albumin did not. IL-1alpha and tumor necrosis factor (TNF)-alpha levels after i.v. injection of CEA were only 3 5% of those induced by LPS. The IL-6 responses of mice pretreated with 100 microg/kg genistein were decreased by more than 40%. However, genistein inhibited the TNF-alpha response to LPS by 46% but increased the CEA-induced response by 300%. When murine Kupffer cells were stimulated with LPS or CEA in vitro, LPS increased tyrosine phosphorylation of a Mr 30,000 protein, whereas CEA decreased phosphorylation of a Mr 60,000 protein and did not increase phosphorylation of the Mr 30,000 protein. Thus, i.v. CEA stimulates production of IL-6 and TNF-alpha after binding to Kupffer cells through signal transduction pathways that appear to be different from those stimulated by LPS. PMID- 9331109 TI - Eighth Annual Pezcoller Symposium: genomic instability and immortality in cancer. PMID- 9331110 TI - Introduction to the treatment of lung cancer. AB - Lung cancer represents the leading cause of cancer mortality worldwide. Its incidence has declined in men but is increasing in women, assuring that this largely preventable disease will continue to affect millions. In small cell lung cancer, the advent of chemotherapy in the 1970s and continued refinements in the 1980s yielded improved median survivals in patients with both limited disease and extensive disease and improved long-term survival in limited-disease patients. Reinduction chemotherapy was shown in the 1980s to improve survival in patients who had achieved a complete remission to initial chemotherapy, and a consensus subsequently developed regarding standard induction chemotherapy for patients with small cell lung cancer. The prognosis for patients with small cell lung cancer depends largely on delivering the optimal combination chemotherapy to achieve early, maximal cell kill with manageable toxicity. Future challenges include comparing newer combinations and novel schedules of administration with "standard" chemotherapy, optimizing the use of complementary treatment modalities, and refining prognostic factors to better define treatment and improve outcome. In patients with non-small cell lung cancer, single-modality treatment has been compared with combined-modality therapy in numerous randomized trials, with consistent survival benefits accrued by patients in combined modality treatment arms. The recent availability of novel cytotoxic and cytostatic agents has prompted additional comparisons of new combination chemotherapy regimens, with or without other treatment modalities, in patients with lung cancer. Questions for the future include defining the most effective chemotherapy to eradicate distant metastases and understanding which modalities offer superior local control. PMID- 9331111 TI - Single-agent paclitaxel in advanced non-small cell lung cancer: single-center phase II study using a 3-hour administration schedule. AB - The efficacy and safety of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) administered as a 3-hour infusion was investigated in a phase II study involving 21 patients with stage III/IV non-small cell lung cancer. The study included two quality of life assessments (the Hospital Anxiety and Depression Scale and the Rotterdam Symptom Checklist) to test their suitability for use in a future randomized phase III trial of paclitaxel and best supportive care versus best supportive care alone. Four (19%) of the 21 patients (95% confidence interval, 8% to 38%) achieved a partial response. The median time to disease progression for all patients entered was 19 weeks. Paclitaxel was well tolerated, with dose reduction required in only one patient because of arthralgia/myalgia. No dose reductions, delays, or discontinuations were required for hematologic toxicity. Completion and compliance with quality of life questionnaires was high, and these research tools proved to be acceptable for future use in phase III studies with paclitaxel. PMID- 9331112 TI - Weekly paclitaxel in patients with advanced lung cancer: preliminary data from a phase II trial. AB - We conducted a phase II trial in chemotherapy-naive patients with advanced non small cell lung cancer to determine the efficacy of paclitaxel (Taxol; Bristol Myers Squibb Company, Princeton, NJ) delivered at a maximum tolerated dose of 175 mg/m2 on an extended weekly schedule. Patients with stage IIIB/IV non-small cell lung cancer were eligible if they had an Eastern Cooperative Oncology Group performance status of 0 to 2, had received no previous chemotherapy, demonstrated normal hematologic and hepatic function, and could provide informed consent. Paclitaxel 175 mg/m2 was administered as an intravenous infusion weekly over 3 hours with standard premedication, for 6 weeks of an 8-week cycle. Doses were modified for absolute neutrophil count less than 1.5 x 10(9)/L or neuropathy greater than grade I on the day of therapy. Patients without progressive disease received subsequent cycles at the same dose. To date, 30 patients have been enrolled; data are available for 25. The median age was 65 years (range, 38 to 78 years), 23 patients were performance status 0 or 1, and 14 had received prior radiation. Sites of disease included the lung (23 patients), central nervous system (11), bone (seven), liver (one), kidney (one), and soft tissue (eight). Eighty-three percent, 75%, 58%, and 50% of intended doses were delivered during cycles 1 though 4, respectively. Grade 2/3 neuropathy occurred in nine patients, but improved in all nine following dose reduction. Grade 3/4 neutropenia occurred in 10 patients. Partial responses occurred in 14 of 25 patients (56%; 95% confidence interval, 46% to 66%). Median duration of response was 6.5 months, and the 1-year survival rate was 53%. The extended weekly paclitaxel schedule results in enhanced dose intensity, marked activity, and acceptable toxicity. Paclitaxel given weekly at maximum dose intensity may be more effective than conventional paclitaxel administration schedules. PMID- 9331113 TI - Paclitaxel (1-hour infusion) plus carboplatin in the treatment of advanced non small cell lung cancer: results of a multicenter phase II trial. AB - This study was performed to determine the activity and toxicity of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) given by 1-hour infusion plus carboplatin in the treatment of patients with advanced non-small cell lung cancer when used in a multicenter, community-based setting. The study population included 100 chemotherapy-naive patientswith stage IIIB or IV non-small cell lung cancer, Karnofsky performance status 70 to 100, measurable disease, and adequate kidney, liver, and bone marrow function. All patients received paclitaxel 225 mg/m2 intravenously by 1-hour infusion followed immediately by carboplatin at a targeted area under the concentration-time curve of 6.0 (Calvert formula). Cycles were repeated every 21 days. Colony-stimulating factors were not used routinely. Thirty-eight (38%) of 100 patients had objective responses (38 [40%] of 94 evaluable patients) to treatment (three complete responses, 35 partial responses). Thirty-two other patients had stable disease at initial re evaluation. Weight gain during treatment occurred in 47% of those patients with objective response or stable disease. The median survival among all 100 patients was 8 months, with a 1-year survival rate of 42%. Leukopenia was common, but hospitalization for treatment of neutropenia and fever occurred in only 3% of courses. Cumulative peripheral neuropathy occurred consistently, but usually after the third or fourth course; it was severe (grade 3) in only 15% of patients. Other grade 3 and 4 toxicity was uncommon. One patient died as a result of treatment due to sepsis. This large, multicenter, community-based phase II trial demonstrates the efficacy of paclitaxel/carboplatin combination chemotherapy in advanced non-small cell lung cancer. This regimen is relatively well tolerated and when paclitaxel is given by 1-hour infusion, this treatment is easily administered in the outpatient setting. PMID- 9331114 TI - Preliminary results of a phase II study of paclitaxel and cisplatin in patients with non-small cell lung cancer. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and cisplatin are cytotoxic drugs active against non-small cell lung cancer (NSCLC) that possess additive cytotoxicity in animal tumors. Paclitaxel and cisplatin are active in patients with advanced NSCLC when given on a 3-weekly schedule. In an attempt to increase activity, we designed a phase II study with a biweekly schedule. Paclitaxel 110 mg/m2 was given by 3-hour intravenous infusion, followed by cisplatin 60 mg/m2 via intravenous infusion. Treatment was scheduled every 2 weeks. Of the 42 patients treated, 19 were men and 23 were women, with a median age of 54 years (range, 31 to 69 years). Four patients had stage IIIA NSCLC, 18 stage IIIB, and 20 stage IV. Median World Health Organization performance status was 1 (range, 0 to 2), and adenocarcinoma was the most common histology (52%). A median of nine cycles was administered (range, one to 24 cycles), with more than 360 cycles administered. Rates of frequency of World Health Organization grade 3 or 4 toxicities were as follows: neutropenia, 20%; thrombocytopenia, 2%; nausea/vomiting, 7% (despite prophylactic treatment with 5-HT3 receptor antagonists plus prednisolone); neurotoxicity, 2%; and nephrotoxicity, 2%. There were three septicemic episodes, no bleeding episodes, and no toxic deaths. Dose reduction was performed in 15 patients (36%), due to nephrotoxicity in 14 cases. Treatment delay was necessary in 23 patients (55%), most often due to neutropenia (nine cases). Forty patients are currently evaluable for response, with two complete and 15 partial responses (overall response rate, 43%; 95% confidence limits, 27% to 59%). Median response duration was 31 weeks (range, 9 to 85 weeks). The biweekly schedule of paclitaxel plus cisplatin has noteworthy activity in patients with NSCLC. A relatively large fraction of patients required either dose reduction and/or treatment delay, but World Health Organization grade 3 or 4 toxicity was rare, apart from the neutropenia that caused only a few septicemic episodes. PMID- 9331115 TI - Preliminary analysis of a phase II study of paclitaxel, carboplatin, and hyperfractionated radiation therapy for locally advanced inoperable non-small cell lung cancer. AB - We conducted a prospective phase II study to determine the response rate, toxicity profile, and survival rate among patients with locally advanced unresectable non-small cell lung cancer receiving concurrent weekly paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), carboplatin, and hyperfractionated radiation therapy followed by two cycles of adjuvant paclitaxel and carboplatin. The weekly paclitaxel/carboplatin regimen was designed to optimize the radiosensitizing properties of paclitaxel during the concurrent phase of treatment. Thirty-two patients with unresectable stage IIIA and IIIB non small cell lung cancer from Vanderbilt Cancer Center Affiliate Network institutions entered the study from June 1996 until February 1997. Weekly intravenous paclitaxel (50 mg/m2 over 1 hour) and weekly carboplatin (area under the concentration-time curve of 2) plus concurrent hyperfractionated chest radiotherapy (1.2 Gy twice daily [69.6 Gy total]) delivered for 6 weeks were followed by two cycles of paclitaxel (200 mg/m2) and carboplatin (area under the concentration-time curve of 6). Among 22 patients evaluable for response, one (4.5%) achieved a complete response and 16 (72.7%) achieved partial response, for an overall response rate of 77%. Among 23 patients evaluable for toxicity, esophagitis was the principal finding: grade 3 or 4 esophagitis occurred in eight patients (35%). Grade 3 and 4 pulmonary toxicities each occurred in 26% of patients. Thus, weekly paclitaxel/carboplatin plus concurrent hyperfractionated radiotherapy is a well-tolerated outpatient regimen with an encouraging response rate that is at least equivalent to more toxic chemoradiation regimens. These findings indicate that further clinical evaluation of weekly paclitaxel/carboplatin/hyperfractionated radiotherapy is warranted in phase III trials. PMID- 9331116 TI - Paclitaxel by either 1-hour or 24-hour infusion in combination with carboplatin in advanced non-small cell lung cancer: preliminary results comparing sequential phase II trials. AB - Our group previously described the activity of carboplatin plus paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) (given as a 24-hour infusion) in 51 patients with advanced non-small cell lung cancer. To facilitate outpatient administration, the regimen was modified to infuse paclitaxel over 1 hour. Between February 1995 and August 1996, 63 patients with advanced non-small cell lung cancer were accrued by the Vanderbilt Cancer Center and its affiliate network. The first four patients received paclitaxel 175 mg/m2; all subsequent patients received paclitaxel 200 mg/m2. The carboplatin dose was determined using the Calvert formula, with a target area under the concentration-time curve of 6. Cycles were repeated every 4 weeks, to a maximum of six cycles. The median age of the patients was 62 years. There were 43 men and 20 women. Ten patients were stage IIIB and 53 were stage IV. Patients with Eastern Cooperative Oncology Group performance status < or =2 were enrolled. There were three complete remissions and 13 partial remissions, for an overall response rate of 25%. Median survival was 32 weeks. This compares with a response rate of 27% and a median survival of 38 weeks observed in our previous study, using 24-hour paclitaxel plus the same dose of carboplatin. Grade 3/4 leukopenia occurred in 47% versus 3% of treatment cycles in the 24-hour versus 1-hour patient groups, respectively. Febrile neutropenia was similar and occurred in 7% versus 4% of treatment cycles. Grade 1 to 3 neurotoxicity occurred in 7% versus 41% of patients in the 24-hour versus 1 hour schedule groups, respectively. Likewise, the incidence of grade 1 to 3 arthralgia/myalgia was greater among patients receiving 1-hour infusion of paclitaxel (3.5% v 28%). Although not randomized, these data suggest that survival may be comparable whether paclitaxel is given by short or prolonged infusion in advanced non-small cell lung cancer. Toxicity profiles differ, however, with greater myelosuppression following 24-hour paclitaxel, but a higher incidence of neurotoxicity and arthralgia/myalgia with the 1-hour infusion. PMID- 9331117 TI - Paclitaxel (175 mg/m2) plus carboplatin versus paclitaxel (225 mg/m2) plus carboplatin in non-small cell lung cancer: a randomized study. AB - A recent phase II study by our group documented a response rate of 27% with the combination paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) 175 mg/m2 plus carboplatin dosed to a target area under the concentration-time curve of 7 in patients with advanced non-small cell lung cancer. In an effort to evaluate the dose-response relationship of paclitaxel with quality of life, we initiated a phase III prospective trial. Patients with inoperable non-small cell lung cancer were randomized into two groups. Group A received paclitaxel 175 mg/m2 plus carboplatin dosed to an area under the concentration-time curve of 6 every 3 weeks. Group B received the same regimen, with paclitaxel increased to 225 mg/m2. Since July 1996, 49 patients have entered the study, 29 in group A and 20 in group B. Patient and tumor characteristics were well distributed between both groups. In group A, 16 patients were evaluable, with one complete response, six partial responses, three stable disease, and six progressive disease. In group B, 12 patients were evaluable, with two partial responses, four stable disease, and six progressive disease. Treatment was well tolerated in both groups. More neurotoxicity and neutropenia were noticed with high-dose paclitaxel. There were no drug-related deaths. It is too early to draw definite conclusions regarding response and survival, but regarding toxicity, it seems that paclitaxel 225 mg/m2 plus carboplatin dosed to an area under the concentration-time curve of 6 is well tolerated without the use of growth factors. Although the results are premature, quality of life does not seem to be affected by the increased paclitaxel dose. PMID- 9331118 TI - Preliminary results of neoadjuvant paclitaxel and carboplatin in the treatment of early stage non-small cell lung cancer. AB - The purpose of this study is to determine the feasibility of delivering neoadjuvant paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and carboplatin to patients with clinical early stage (stage I and II) non-small cell lung cancer. Although neoadjuvant chemotherapy appears to prolong survival in patients with stage IIIA non-small cell lung cancer, several studies have demonstrated an increase in perioperative mortality associated with this approach. This study is designed to address whether three cycles of paclitaxel (200 mg/m2/3 hour, day 1) and carboplatin (area under the concentration-time curve 5, day 2) can be given preoperatively to patients with clinical stage I and II non-small cell lung cancer and to assess the associated toxicities, pathologic response rate, disease-free survival, and overall survival of this group of patients. Thus far, five patients have been enrolled. Three have successfully undergone resection, with no perioperative complications noted. One patient had a pathologic complete remission and two had pathologic partial remissions. Preliminary results indicate that this approach is well tolerated and results in major tumor response. PMID- 9331119 TI - Paclitaxel (3-hour infusion) followed by carboplatin (24 hours after paclitaxel): a phase II study in advanced non-small cell lung cancer. AB - This phase II study was performed to investigate the efficacy of a 3-hour 225 mg/m2 paclitaxel infusion (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) followed 24 hours later by a 30-minute infusion of carboplatin (dosed to an area under the concentration-time curve of 6) in patients with stage IIIA, IIIB, or IV non-small cell lung cancer. Patients received chemotherapy and were monitored for toxicity, response, quality of life, and survival. Paclitaxel and carboplatin pharmacokinetics were also determined with the first cycle of chemotherapy. Eleven men have been treated to date. Eight were white and three black, with a median age of 65 years. All patients had a performance status of 0 or 1. The regimen was well tolerated, with no deaths or grade 4 toxicities noted. The most common grade 3 toxicity was neutropenia, thrombocytopenia, and parasthesias (observed in <10% of cycles). The overall response rate was 57% (14% complete and 43% partial responses). Quality of life improved in most patients. Physical and emotional well-being improved in 57%, functional well-being in 43%, and social/family well-being in 14% of patients. Pharmacokinetic data are being analyzed by limited sampling technique to predict the paclitaxel area under the concentration-time curve. This unique schedule of paclitaxel and carboplatin is well tolerated and active, and is associated with improvements in various aspects of quality of life. PMID- 9331120 TI - Induction paclitaxel/carboplatin in early stage non-small cell lung cancer. Bimodality Lung Oncology Team. AB - In this feasibility study, a 3-hour infusion of 225 mg/m2 paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) was combined with carboplatin dosed to an area under the concentration-time curve of 6 to treat patients with stage T2N0, T1-2N1, or T3N0-1 (excluding superior sulcus tumors) non-small cell lung cancer. Nineteen of a planned 80 patients have been enrolled. To assure that patients meet the study's criteria for inclusion, rigorous physical and laboratory investigations are performed before, during, and after the preoperative chemotherapy and again before the postsurgical chemotherapy. Treatment includes two cycles of preoperative chemotherapy, followed within 3 to 6 weeks with thoracotomy. Up to three cycles of postoperative chemotherapy are planned, commencing 3 to 8 weeks after surgery, as tolerated. To date, 14 patients have completed induction chemotherapy, nine of whom have undergone surgical resection. Three patients have completed postoperative chemotherapy. The study treatment has been well tolerated with no unexpected toxicities. Very preliminary results suggest that perioperative paclitaxel/carboplatin appears to be a feasible and tolerable regimen in patients with early stage non-small cell lung cancer and warrants further investigation. More mature results may provide the basis for an intergroup randomized trial comparing this regimen with surgery alone for patients with early stage disease. PMID- 9331121 TI - High-dose therapy with carboplatin and paclitaxel in non-small cell lung cancer. AB - Non-small cell lung cancer (NSCLC) remains the leading cause of cancer deaths in the United States. The combination of more active agents like vinorelbine and paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) with cisplatin has led to improved survival for patients with advanced metastatic disease. The ability to escalate the dose of cisplatin-based regimens is limited by nonhematologic toxicities and is especially difficult in the population of patients with advanced NSCLC. However, the development of new agents with significant activity against NSCLC as well as new strategies for high-dose therapy have made it possible to examine the potential of dose-intense therapy in this population. A phase I trial performed at the University of North Carolina was designed to evaluate paclitaxel 250 mg/m2 given over 24 hours plus escalating doses of carboplatin starting at an area under the concentration-time curve (AUC) dose of 8 supported by peripheral blood stem cells and filgrastim in the treatment of advanced NSCLC. The AUC dose of carboplatin was escalated in increments of 2. The maximum tolerated dose of carboplatin combined with paclitaxel 250 mg/m2 administered over 24 hours was defined at an AUC of 18. In this study, six of seven patients with advanced NSCLC had major responses. This regimen is currently being tested in patients with locally advanced NSCLC by the Cancer and Leukemia Group B: patients receive two cycles of induction therapy with paclitaxel 250 mg/m2 over 3 hours followed by carboplatin at an AUC of 18 supported by peripheral blood stem cells and filgrastim. Depending on response to induction therapy, patients then receive surgical resection, thoracic radiation therapy, or both. This phase II trial will examine clinical and pathologic responses and the toxicity of this high-dose regimen in patients with locally advanced NSCLC. Ultimately, phase III trials will be needed to establish the role of this approach in NSCLC. PMID- 9331122 TI - Paclitaxel/carboplatin chemotherapy as primary treatment of brain metastases in non-small cell lung cancer: a preliminary report. AB - Chemotherapy has been rarely considered an important treatment modality for brain metastases. Based on the hypothesis that the lack of efficacy of chemotherapy, rather than the blood-brain barrier itself, may be the major hindrance to the successful chemotherapeutic treatment of brain metastases, we started a trial in which a selected group of non-small cell lung cancer patients with brain metastases received primary treatment with systemic chemotherapy. The treatment consisted of three courses of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) 225 mg/m2 given intravenously over 3 hours and carboplatin dosed to an area under the concentration-time curve of 6, with close monitoring of the lesion by computed tomography or magnetic resonance imaging of the brain after each chemotherapy course. Any radiographic or clinical evidence of progression in the brain during treatment or no improvement in the brain after three cycles of chemotherapy mandated whole brain irradiation (30 Gy in 10 fractions). Responding patients received three additional courses of chemotherapy; whole brain irradiation was given after completion of all six chemotherapy cycles. To date, five patients have been enrolled, and one has achieved partial remission both in the brain and at the extracranial site. Other patients did not achieve major objective responses either in the brain or at the extracranial sites. These preliminary results, which are consistent with the study hypothesis, support the feasibility of our approach. We therefore continue to accrue patients for this study. PMID- 9331123 TI - A sequence-dependent paclitaxel/etoposide phase II trial in patients with non small cell lung cancer. AB - Studies conducted by the Spanish Lung Cancer Group indicate that cisplatin- or carboplatin-based chemotherapy can yield a 25% response rate, 9-month median survival time, and 30% 1-year survival rate in patients with stage III and IV non small cell lung cancer. Phase II trials of single-agent paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) have an almost 30% response rate in non-small cell lung cancer. Based on these results, we decided to examine whether the sequence-dependent effects of paclitaxel/etoposide influence treatment outcome (antitumor response) and toxicity. In vitro data show a paradoxical antagonist rather than additive effect. In the first part of our study (part A), paclitaxel and etoposide were administered at the same time. In the second part (part B), etoposide preceded paclitaxel. In both parts, patients with previously untreated stage IIIB or IV non-small cell lung cancer with good performance status were eligible. In part A, etoposide (fixed dose, 100 mg/m2) on days 1, 2, and 3 was administered by 30-minute infusion; paclitaxel (175 mg/m2) was given by a 3-hour infusion on day 1. In part B, the etoposide dose and schedule were the same, but paclitaxel (same dose) was administered on day 4. Treatment in both parts was repeated every 21 days for a maximum of 10 cycles. In part A, 18 patients were entered and no objective responses were observed. In part B, 21 patients were accrued, 17 of whom had sufficient follow-up for response assessment. Seven objective responses were achieved (two complete and five partial responses, for an objective response rate of 41%). Seven patients had no change and three had progressive disease. Frequency and severity of side effects were not significantly different in either part of the study. However, grade 4 neutropenia was observed in 10 (59%) patients and one (5%) patient in parts A and B of the trial, respectively. Nonhematologic toxicity was slight. In conclusion, paclitaxel cytotoxicity is abrogated when it is given concurrently with etoposide. When etoposide precedes paclitaxel, a more effective paclitaxel/etoposide schedule is attained. PMID- 9331124 TI - Second-line treatment of advanced non-small cell lung cancer with paclitaxel and gemcitabine: a preliminary report on an active regimen. AB - A phase II study of combination paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ)/gemcitabine was conducted in patients with non-small cell lung cancer (NSCLC) who had failed first-line docetaxel- or cisplatin-based chemotherapy. Eligibility criteria included histologically confirmed measurable stage IIIB or IV NSCLC and previous exposure to docetaxel- or cisplatin-based regimens, World Health Organization performance status between 0 and 2, adequate hematologic parameters, and adequate hepatic, renal, and cardiac function. Gemcitabine (900 mg/m2) was given on days 1 and 8 as a 30-minute infusion; paclitaxel (175 mg/m2) was administered on day 8 as a 3-hour infusion after appropriate premedication. Granulocyte colony-stimulating factor (150 microg/m2 subcutaneously) was given on days 9 to 15. Treatment was repeated every 3 weeks until patients experienced disease progression. From October 1995 to December 1996, 26 patients with advanced NSCLC were enrolled (three stage IIIB, 23 stage IV). All 26 patients were assessable for toxicity, and 24 were evaluable for response. Two complete (8%) and five partial (21%) responses were observed, for an overall response rate of 29% (95% confidence interval, 11% to 47%). The median duration of response was 2.5 months and the median survival was 8 months. A median of three courses per patient was administered, and the median interval between courses was 21 days. The median delivered dose was 579 mg/m2/wk gemcitabine and 54.5 mg/m2/wk paclitaxel, corresponding to a relative dose intensity of 0.97 and 0.96, respectively. Grade 3/4 neutropenia occurred in two patients (8%). Grade 3 conjunctivitis occurred in one (4%) patient and grade 2/3 neurotoxicity in eight (31%) patients. Grade 3/4 and grade 2 fatigue occurred in four (15%) and eight (31%) patients, respectively. Other toxicities were mild to moderate. These preliminary results suggest that the paclitaxel/gemcitabine combination is an active and well-tolerated salvage regimen in patients with NSCLC previously treated with docetaxel- or cisplatin-based chemotherapy. The paclitaxel/gemcitabine combination merits further evaluation as first-line treatment. PMID- 9331125 TI - Paclitaxel/carboplatin plus ifosfamide in non-small cell lung cancer. AB - Chemotherapy has a positive role in managing patients with stage IV non-small cell lung cancer. Randomized studies and meta-analyses comparing chemotherapy with best supportive care have confirmed a significant prolongation of survival for chemotherapy-treated patients. In recent years, several new active agents have been identified. These include the taxanes paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and docetaxel, the antimetabolite gemcitabine, the mitotic spindle inhibitor vinorelbine, and (potentially) the topoisomerase I inhibitors. Combinations of either vinorelbine or paclitaxel with cisplatin have resulted in a significant increase in median survival times compared with both cisplatin and vindesine and cisplatin and etoposide. Paclitaxel combined with either carboplatin or ifosfamide also has been promising. Since even the best current combination regimens result in median survival times of less than 1 year, the identification of more active drugs and combinations remains a high priority. One possible strategy to identify more active regimens may be the combination of three rather than two active agents into a combination regimen. Although the three-drug regimens used in the 1980s appeared to be no more active than two-drug combinations, the advent of additional active compounds with novel mechanisms of action allows this question to be readdressed. Based on this background, we have initiated a phase I/II study of carboplatin and paclitaxel with escalating doses of ifosfamide. The study design and dosing schedule are discussed in this report. PMID- 9331126 TI - Paclitaxel, ifosfamide, and carboplatin for the treatment of stages IIIB and IV non-small cell lung cancer: preliminary results. AB - We have treated 26 consecutive chemotherapy-naive patients with stage IIIB/IV non small cell lung cancer with an innovative regimen based on a 1-hour infusion of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) 125 to 250 mg/m2, ifosfamide 3 g/m2 (with mesna), and carboplatin dosed to an area under the concentration-time curve of 5, every 21 days for a total of six cycles in responding or stabilized patients. Among 22 fully evaluable patients, 14 (64%) achieved a partial remission, six had disease stabilization, and two had disease progression. Hematologic toxicity was remarkably mild; only one patient had grade 3 neutropenia (on day 21). Arthralgias/myalgias (grade 3 in nine patients, grade 4 in one patient) and neurologic toxicity (a cumulative sensory neuropathy of grade 3 or 4 in five patients) were the most common side effects and seem to be dose related. To date, few patients are fully evaluable and survival data are clearly immature. Nevertheless, this regimen seems highly active and quite well tolerated, and deserves further evaluation. PMID- 9331127 TI - Ifosfamide/carboplatin/etoposide/paclitaxel in advanced lung cancer: update and preliminary survival analysis. AB - The primary objective of this study was to define the maximum tolerated dose and toxicity profile of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), given as a 24-hour infusion, in conjunction with ifosfamide/carboplatin/etoposide (ICE) chemotherapy in patients with advanced lung cancer. Paclitaxel was escalated from 75 to 225 mg/m2 in 25-mg/m2 increments. All patients received granulocyte colony-stimulating factor 5 microg/kg/d from day 4 until the neutrophil count was > or = 10,000/microL. The study population consisted of 41 patients with a median age of 60 years and a median follow-up of 20.7 months. Stage distribution included 5% stage IIIA, 46% stage IIIB, and 49% stage IV. Histology consisted of 61% adenocarcinoma, 12% squamous cell carcinoma, 10% large cell carcinoma, 15% small cell carcinoma, and 2% mixed. The predominant toxicity was hematologic; 63% of patients experienced grade 4 neutropenia and 49% developed grade 4 thrombocytopenia. Fever and neutropenia occurred in 34% of patients. Hematologic toxicity was, in all cases, short-term and reversible and was not dose related. With few exceptions, nonhematologic toxicity was not clinically important. Among 39 patients evaluable for response, 36% achieved a remission (8% complete, 28% partial, 41% had stable disease, and 23% experienced disease progression). Among 33 patients with non small cell lung cancer, the response rate was 27% (one complete response, eight partial responses, 15 had stable disease, and nine had progressive disease). Among six patients with small cell carcinoma, the response rate was 83% (two complete responses, three partial responses, and one had stable disease). The median survival of all 41 patients was 13.6 months. Survival was almost identical between stage IIIA and stage IV subsets. We conclude that it is possible to safely administer full-dose single-agent paclitaxel with granulocyte colony stimulating factor support in conjunction with full-dose ifosfamide/carboplatin/etoposide chemotherapy. While response rates observed were not particularly notable, median survival is considerably longer than that usually achieved with combination chemotherapy in advanced lung cancer. PMID- 9331128 TI - Paclitaxel (1-hour) and carboplatin (area under the concentration-time curve 7.5) in advanced non-small cell lung cancer: a phase II study of the Fox Chase Cancer Center and its network. AB - We previously reported a 62% response rate and 54% 1-year survival rate for paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) administered by 24-hour infusion in combination with fixed-dose carboplatin to treat patients with advanced non-small cell lung cancer (NSCLC). Myelosuppression proved dose limiting, but was substantially reduced by the routine use of granulocyte colony stimulating factor during the second and subsequent cycles. Activity for paclitaxel 135 mg/m2 and 200 mg/m2 by 1-hour infusion every 3 weeks in patients with NSCLC, with minimal myelosuppression and the suggestion of a dose-response relationship, has been reported. In November 1994, we initiated a phase II trial in patients with advanced, measurable, chemotherapy-naive NSCLC using paclitaxel 175 mg/m2 given in 1 hour, and carboplatin dosed to a fixed target area under the concentration-time curve of 7.5 every 3 weeks. In the absence of grade 4 myelosuppression, paclitaxel was escalated on an intrapatient basis by 35 mg/m2 per cycle to a maximum dose of 280 mg/m2 by cycle 4. Granulocyte colony stimulating factor was not routinely used. Of the 57 patients accrued, 44 (81%) are Eastern Cooperative Oncology Group performance status 1. The median patient age is 64 years. To date, 54 patients are fully evaluable for toxicity. In the first 20 evaluable patients accrued (cohort A), myelosuppression was tolerable, but cumulative peripheral sensory neuropathy proved dose limiting: grade > or = 1 in 15 (75%) patients and grade 3 in six (30%), generally occurring at paclitaxel doses > or = 215 mg/m2 and obligating at least three patients to be removed from study despite absence of disease progression. The protocol was consequently revised. The starting dose of paclitaxel was reduced to 135 mg/m2 with intrapatient dose escalations of 40 mg/m2 per cycle, to a maximum paclitaxel dose of 215 mg/m2, recapitulating the original dosing schema used in Fox Chase Cancer Center study 93-024. For the 35 patients enrolled in the second cohort (cohort B), treatment has been better tolerated. Of 21 evaluable patients, 13 (62%) have experienced peripheral sensory neuropathy, grade 3 in only one (5%) patient. Myelosuppression also has been less pronounced, with 44% grade 4 granulocytopenia and 38% grade > or =3 thrombocytopenia in cohort B compared with 70% and 50%, respectively, in cohort A. Of the first 22 patients accrued to cohort A, 12 (55%) had major objective responses. Median event-free survival is 24 weeks and median survival is 47 weeks. Of the 35 evaluable patients in cohort B, nine (26%) have had major objective responses. Median event-free survival is 22 weeks. It is too early to report median survival. Paclitaxel given by 1-hour infusion in combination with carboplatin at a fixed target area under the concentration-time curve of 7.5, although active in advanced NSCLC, is associated with problems that compromise its efficacy. Higher dose levels yield intolerable toxicity, evidenced by the incidence of neurotoxicity (rather than myelosuppression) that was dose and protocol limiting at paclitaxel doses exceeding 215 mg/m2. Lower doses, while more tolerable, appear to be associated with lower response rates. PMID- 9331129 TI - Induction paclitaxel and carboplatin followed by concurrent chemoradiotherapy in patients with unresectable, locally advanced non-small cell lung carcinoma: report of Fox Chase Cancer Center study 94-001. AB - The paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ)/carboplatin combination has demonstrated promising activity in patients with incurable non small cell lung cancer (NSCLC). Our exploratory study is designed to evaluate the efficacy of this combination as induction therapy in patients with locally advanced NSCLC, to determine the maximally tolerated doses of paclitaxel and carboplatin administered every 3 weeks during radical thoracic radiation after induction treatment, and to determine the efficacy of granulocyte colony stimulating factor (G-CSF) priming before induction treatment, followed by conventional G-CSF, compared with conventional G-CSF alone. Eligibility stipulated Karnofsky performance status > or =70%, < or =5% weight loss, and stages IIIB or bulky IIIA NSCLC. Induction treatment consisted of two cycles of paclitaxel 175 to 225 mg/m2 infused over 3 hours combined with carboplatin (target area under the concentration-time curve of 7.5) given on days 1 and 22. On days 2 through 15 and 23 through 36, all patients received G-CSF 5 microg/kg; half were randomized to receive priming G-CSF daily for 5 days before day 1 of treatment. On day 43, thoracic radiation (60 Gy in 30 2-Gy fractions daily, 5 days a week for 6 weeks) was initiated. At dose level 1, patients received carboplatin dosed to a target area under the concentration-time curve of 3.75 and paclitaxel 67.5 mg/m2 over 3 hours on days 43 and 64. In the absence of dose limiting toxicity, phase I escalation in three-patient cohorts proceeded to a maximum carboplatin area under the concentration-time curve of 5.0 and a paclitaxel dose of 175 mg/m2, delivered over 3 hours. To date, 35 patients (83% stage IIIB) have received induction treatment, 29 of whom are evaluable for response. Myelosuppression and neurotoxicity have been mild during induction treatment, prompting a paclitaxel dose increase to 225 mg/m2 on days 1 and 22 after the first seven patients were accrued. The phase III portion of the study evaluating G-CSF priming remains coded. Sixteen patients have received concurrent thoracic radiation and chemotherapy and are evaluable for response and toxicity. In sequential cohorts, the paclitaxel dose on days 43 and 64 has been escalated to 175 mg/m2 with only one episode each of grade 4 granulocytopenia and grade 3 anemia. In the first 13 patients evaluated, the severity of esophagitis corresponded to the length of the esophagus in the radiation treatment field: grade 1 in all six patients with esophageal exposure < or =16 cm and grade > or =2 in six of seven patients with > or =16 cm of the esophagus irradiated. Three episodes of grade > or =2 steroid-responsive pulmonary toxicity have occurred 2 to 6 months after the conclusion of concurrent thoracic radiation and chemotherapy. The major response rate is 38% to induction treatment and 59% to combined-modality treatment. Of the first 21 patients accrued, 62% survived 1 year. Induction paclitaxel/carboplatin therapy is active and well tolerated by patients with locally advanced NSCLC. The maximum tolerated doses of paclitaxel and carboplatin during concurrent thoracic radiation and the role of G-CSF priming are not yet established. Severity of esophagitis corresponds to the extent of esophagus irradiated during concurrent thoracic radiotherapy and chemotherapy. PMID- 9331130 TI - Seven-week continuous-infusion paclitaxel plus concurrent radiation therapy for locally advanced non-small cell lung cancer: a phase I study. AB - The goal of this National Cancer Institute-sponsored phase I trial is to determine the feasibility, toxicity, and pharmacokinetics of continuous-infusion (24 hr/d, 7 d/wk, 7 weeks total) intravenous paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) when combined with standard, curative-intent radiation therapy (RT) for previously untreated, locally advanced non-small cell lung cancers. Eligible patients have locally advanced (T4NXM0 or TXN2-3M0) non small cell cancer ineligible for potentially curative surgical resection, a good performance status, adequate hematologic, hepatic, and renal functions, and no distant metastases. All patients receive a total tumor dose of 64.8 Gy megavoltage RT in 7 weeks at 1.8 Gy once daily, 5 d/wk. Paclitaxel is delivered by continuous intravenous infusion starting 48 hours before RT and continuing for its duration. The dose of paclitaxel is escalated in cohorts of three patients in a standard phase I design. To date, 16 patients have entered the trial, and 15 are evaluable for toxicity in this ongoing study. Paclitaxel dose is currently at a 6.5 mg/m2/d dose level, with no dose-limiting toxicity recorded thus far. One patient at the highest dose level has had grade 2 pneumonitis. With the exception of anemia, toxicities are those that would be expected from RT alone. A slowly progressive normocytic anemia with no renal dysfunction was found to be associated with an acquired hypoerythropoietin state. These findings indicate that this therapy is feasible and well tolerated through current dose levels, with no dose-limiting toxicity. Dose escalation is ongoing. PMID- 9331131 TI - Simultaneous paclitaxel and radiotherapy: initial clinical experience in lung cancer and other malignancies. AB - This report summarizes results from a series of pilot trials using combined modality chemoradiotherapy with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) as a radiosensitizing agent in patients with cancers of the lung, cervix, and bladder. In a phase I study of paclitaxel/radiotherapy in patients with locally advanced non-small cell lung cancer, five paclitaxel dose levels were evaluated in conjunction with simultaneous radiation (total dose, 59.4 Gy). A minimum of five patients were treated at each dose level; paclitaxel doses ranged from 45 mg/m2 over 3 weeks (level 1) to 65 mg/m2 for 7 weeks. Of 34 enrolled patients, 25 are evaluable for toxicity and response. Side effects were generally moderate for this combined-modality therapy, although two patients at level 5 developed dose-limiting toxicities (grade 4 esophagitis and grade 3 pneumonitis). Among 25 evaluable patients, complete and partial response rates were 4% (one patient) and 64% (16 patients), respectively; eight patients had a minor response. Median survival was 6 months (range, 1 to 20 months). Therapy was well tolerated, suggesting that the combined modalities offer a practical, effective therapy for patients with non-small cell disease. A paclitaxel dose of 55 mg/m2 is recommended for further study of combined-modality chemoradiotherapy in this clinical setting. In another trial, 33 women with inoperable, locally advanced cervical cancer received carboplatin 50 mg/m2 via intravenous infusion simultaneously with external-beam radiation therapy and vaginal brachytherapy, to define the regimen's toxicity and safety. Among the 33 women, 78% achieved a complete response to therapy. The investigators next conducted a trial of paclitaxel 50 mg/m2 given weekly over 3 hours with the previous carboplatin/radiotherapy regimen in four women and documented two partial responses, one near-complete response, and one minor response, with moderate, manageable toxicity. In a final case report on a patient with recurrent bladder cancer, simultaneous radiotherapy and weekly paclitaxel 50 mg/m2 intravenously over 3 hours yielded a partial remission, prompting the investigators to plan a phase I study to confirm the regimen's efficacy and safety. Additional planned studies include a phase I trial of simultaneous chemoradiotherapy in patients with cancer of the head and neck. PMID- 9331132 TI - Twice-weekly paclitaxel and radiation for stage III non-small cell lung cancer. AB - A phase I study was conducted to investigate the safety and efficacy of twice weekly paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and concurrent thoracic irradiation in patients with stage III non-small cell lung cancer. Radiation therapy beginning on day 1 was delivered in 1.8- to 2.0-Gy daily fractions, to a total dose of 61 Gy. Paclitaxel at a starting dose of 25 mg/m2/d was administered intravenously over 1 hour before daily radiation on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, and 39, for a total of 12 doses over 6 weeks. The paclitaxel dose was escalated by 5 mg/m2/d in each cohort of patients to determine the maximum tolerated dose. The highest paclitaxel dose reached was 40 mg/m2/d, as defined by dose-limiting toxicities of esophagitis and desquamation within the radiation fields. For each dose group, the median total number of paclitaxel doses administered was 12 and the median total radiation dose was 61 Gy. Response rates ranging from 50% to 100% were observed (three of six patients at paclitaxel 25 mg/m2, four of six at 30 mg/m2, seven of seven at 35 mg/m2, six of six at 40 mg/m2), for an overall response rate of 80%. We conclude that the maximum tolerated dose of paclitaxel is 35 mg/m2 given twice weekly in a 1-hour infusion for 6 weeks concurrently with thoracic irradiation. This study provides the basis for an ongoing trial combining twice-weekly paclitaxel and carboplatin with concurrent thoracic irradiation for patients with stage III non-small cell lung cancer. PMID- 9331133 TI - Chemoradiotherapy for poor-risk stage III non-small cell lung cancer. AB - Cisplatin-based chemoradiotherapy is becoming a standard treatment for patients with stage III non-small cell lung cancer (NSCLC). However, a significant proportion of patients with lung cancer also present with co-morbid conditions that indicate a poor prognosis and poor tolerance of treatment. We have completed a phase I/II study to evaluate the tolerability and efficacy of carboplatin-based chemoradiotherapy for patients with poor-risk stage III NSCLC. Twenty-four patients with stage IIIA/B NSCLC and concurrent medical conditions rendering them ineligible for cisplatin-based chemoradiotherapy protocols were treated with thoracic irradiation, 1.8 to 2 Gy daily to the primary tumor and regional lymph nodes, for a total dose of 61 Gy. Concurrently, patients received carboplatin 200 mg/m2/d intravenously on days 1, 3, 29, and 31, and etoposide 50 mg/m2/d intravenously on days 1 through 4 and 29 through 32. Among 23 assessable patients, 96% completed the two planned courses of chemotherapy and 87% completed the planned chest irradiation. Grade 3/4 toxicities included neutropenia in nine patients (39%), thrombocytopenia in five (22%), esophagitis in seven (30%), and nausea in two (9%). Four patients (17%) achieved a complete response and 16 (70%) a partial response, yielding an overall response rate of 87%. The median survival was 12 months, and the 2- and 3-year survival rates were 30% and 20%, respectively. In conclusion, this treatment regimen was relatively well tolerated, with promising response and survival in patients with poor-risk stage III NSCLC. This pilot study provides a basis for further investigation of this treatment regimen. PMID- 9331134 TI - Weekly paclitaxel/cisplatin with concurrent radiotherapy in patients with locally advanced non-small cell lung cancer: a phase I study. AB - We designed a phase I study to determine the maximum tolerated doses of weekly cisplatin and paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) (doses escalated alternately) when given concurrently with standard or hyperfractionated radiotherapy (RT) and to define the nature of the dose-limiting toxicity. Chemotherapy-naive patients with locally advanced non-small cell lung cancer received weekly combination cisplatin/paclitaxel with concurrent local RT. Radiation therapy was initially given at the dose of 1.2 Gy twice daily x 5 d/wk x 5 weeks (total dose, 60 Gy). In the last two patient cohorts, the single daily dose was decreased to 2 Gy x 5 d/wk x 6 weeks. Overall, 25 patients were recruited into five different cohorts. Esophagitis was the main nonhematologic toxicity, occurring in 16 of 25 patients (64%; grade 3 or 4 in five). Neutropenia was the most prevalent hematologic toxicity, occurring in 33 of 141 weekly courses, but grade 4 neutropenia was seen in only four courses. Cisplatin/paclitaxel doses of 35 mg/m2/wk and 45 mg/m2/wk, respectively, were safe when standard RT was used, while the cisplatin dose had to be decreased to 30 mg/m2/wk in patients receiving bifractionation. Two complete and 13 partial responses were observed, for a 60% overall response rate (95% confidence interval, 39% to 79%). Median survival was 16 months, with a 66% 1-year actuarial probability. We thus conclude that the cisplatin/paclitaxel combination given weekly can be safely administered concurrent with both standard or hyperfractionated RT. Hyperfractionation is associated with a higher incidence of severe esophagitis and required a slight reduction in cisplatin dose. To verify whether the use of a daily schedule translates into a better therapeutic index, a new phase I study is under way, testing twice-daily cisplatin/paclitaxel concurrently with hyperfractionated RT. PMID- 9331135 TI - Induction therapy with carboplatin/paclitaxel followed by concurrent carboplatin/paclitaxel and dose-escalating conformal radiotherapy in the treatment of locally advanced, unresectable non-small cell lung cancer: preliminary report of a phase I trial. AB - Locally advanced non-small cell lung cancer is optimally managed with chemotherapy and thoracic irradiation, although the most appropriate strategy is not yet defined. In this phase I trial, we use two 21-day cycles of induction chemotherapy with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) (225 mg/m2 over 3 hours) and carboplatin (area under the concentration-time curve = 6) followed by concurrent weekly paclitaxel (45 mg/m2/wk x 6) and carboplatin (area under the concentration-time curve = 2/wk x 6) and thoracic irradiation. Patients undergo three-dimensional treatment planning (conformal radiotherapy) to define the cancer target volume precisely. The phase I question being addressed in this study is the maximum tolerated radiation dose given concurrently with low dose paclitaxel and carboplatin. The initial radiation dose is 60 Gy, with dose escalations to 66 Gy, 70 Gy, and 74 Gy being planned. Ten patients have been entered thus far (eight men and two women). Their median age is 67 years (range, 59 to 78 years), and none of the patients has had greater than 5% pretreatment weight loss. Seven of 10 are evaluable for response to induction carboplatin and paclitaxel, with a response rate of 57% (three partial responses and one minor response). Three patients had stable disease and none of the patients had evidence of progressive disease during induction chemotherapy. Three patients have completed all treatment at 60 Gy and one has completed all treatment at 66 Gy. Three of the four patients have had partial responses (75%), with the remaining patient having stable disease. Toxicity in the concurrent chemoradiotherapy portion of the trial thus far has consisted of grade 3 neutropenia in one patient and grade 4 lymphocytopenia in all four patients. No grade 3 or 4 nonhematologic toxicity has been seen. The trial data are not yet mature enough to report on survival. Accrual and treatment is continuing at the 66 Gy radiation dose level. PMID- 9331136 TI - Postoperative bronchopulmonary complications in stage III lung cancer patients treated with preoperative paclitaxel-containing chemotherapy and concurrent radiation. AB - We previously observed encouraging results and acceptable toxicity in phase II trials testing preoperative split-course thoracic radiation and simultaneous cisplatin, etoposide, and 5-fluorouracil in stage III non-small cell lung cancer patients. We decided to delete 5-fluorouracil and to incorporate paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) into our combined-modality treatment. The first group of patients received carboplatin dosed at an area under the concentration-time curve of 4 on day 2, etoposide 50 mg orally days 1 to 5 and 8 to 12, cisplatin 50 mg/m2 on day 21, and paclitaxel 35 mg/m2 escalated to 45 mg/m2 on days 1 and 8. Group 2 patients received carboplatin dosed at an area under the concentration-time curve of 4 on day 1, etoposide 45 mg/m2 intravenously daily on days 2 to 5, and paclitaxel 80 mg/m2 (escalating to 120 mg/m2) on day 1. Patients in group 3 received carboplatin dosed at an area under the concentration-time curve of 4 on day 1 and paclitaxel 120 mg/m2 (escalating to 140 mg/m2) on day 1. Each patient received radiation 2 Gy daily on days 1 to 5 and 8 to 12, and a total of two cycles was given at 28-day intervals. Twenty-one patients received preoperative chemoradiotherapy: group 1, five patients; group 2, 11 patients; and group 3, five patients. Thoracotomy was not done in five patients due to cerebrovascular accident in one and progressive tumor in four. The remaining 16 patients had the following procedures: pneumonectomy, eight; lobectomy, six; chest wall resection, one; and no resection, one. Postoperative complications included bronchopleural fistula in one patient each in groups 1 and 3, hypoxia in one patient in group 1, pulmonary hypertension in one patient in group 2, pneumonia in one patient in group 2, and adult respiratory distress syndrome in one patient in group 3, which proved lethal. Thus, six of 16 patients had serious postoperative complications. The relatively high incidence of postoperative bronchopulmonary complications suggests that the use of preoperative paclitaxel-containing chemotherapy and simultaneous thoracic radiation may not be feasible. PMID- 9331137 TI - A phase II study evaluating the efficacy of carboplatin, etoposide, and paclitaxel with granulocyte colony-stimulating factor in patients with stage IIIB and IV non-small cell lung cancer and extensive small cell lung cancer. AB - We initiated a phase II pilot study to determine whether adding paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) to combination carboplatin/etoposide is tolerable and active in patients with advanced non-small cell lung cancer and extensive small cell lung cancer. Patients were given carboplatin (area under the concentration-time curve of 6) followed by etoposide 80 to 100 mg/m2 intravenously on days 1 through 3 followed by paclitaxel 200 mg/m2 intravenously over 3 hours on day 3. On days 4 through 18, granulocyte colony-stimulating factor 5 microg/kg was administered subcutaneously. Each cycle was repeated every 21 days. Fourteen patients have been accrued to the study and 12 were evaluated for toxicity, the first 10 of whom were treated with 80 mg/m2 etoposide. Among the first 10 evaluable patients, significant grade 4 neutropenia occurred in one patient, grade 4 thrombocytopenia in three patients, grade 2 neuropathy in two patients, and grade 3 neurotoxicity in two patients. None of the four patients with non-small cell lung cancer responded to treatment, while six of seven small cell lung cancer patients have obtained major responses to therapy. We have increased the etoposide dose to 100 mg/m2 in subsequent patients. The combination chemotherapy regimen of carboplatin, etoposide, and paclitaxel is tolerable and active in patients with small cell lung cancer. PMID- 9331138 TI - A randomized study of etoposide and carboplatin with or without paclitaxel in the treatment of small cell lung cancer. AB - Small cell lung cancer accounts for 20% to 25% of all lung cancer cases and is initially responsive to combination chemotherapy. However, the majority of patients relapse, and at that point their disease is highly resistant to chemotherapy. The combination of etoposide with either cisplatin or carboplatin is regarded as the standard of care for these patients. Previous studies have documented the activity of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) at doses of 135 to 250 mg/m2 administered over 1, 3, or 24 hours as either a single agent or in combination with etoposide and a platinum compound. Studies adding paclitaxel to etoposide/carboplatin (EP) have demonstrated complete responses in both limited and extensive disease, but all have been in single-arm phase II studies. Preliminary data also suggest the possibility of a dose-response curve for the combination. We recently began a randomized phase II/III comparison of the standard EP to EP plus paclitaxel for newly diagnosed patients with limited or extensive small cell lung cancer. Carboplatin in this study is dosed according to area under the concentration-time curve as calculated by the Calvert formula. The study compares EP (carboplatin area under the concentration-time curve of 6 intravenously [IV] over 30 to 60 minutes on day 1, with etoposide 120 mg/m2 IV days 1 to 3) versus EP plus paclitaxel (paclitaxel 200 mg/m2 IV 1-hour infusion on day 1, carboplatin area under the concentration-time curve of 6 IV over 30 to 60 minutes on day 1, and etoposide 50/100 mg orally on alternating days 1 to 10). The design, inclusion criteria, and staging of patients in this study will be presented with initial accrual and patient characteristics. Randomized studies of this type are essential if the true role of this new combination is to be fully evaluated. PMID- 9331140 TI - A phase I study of cisplatin, etoposide, and paclitaxel in small cell lung cancer. AB - This phase I study was designed to determine the maximum tolerated dose of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) with standard doses of cisplatin and etoposide for patients with untreated, extensive-stage small cell lung cancer. Secondary objectives were to determine the toxicities, response rate, response duration, and overall survival in this cohort. Twenty four patients were enrolled into four dose levels. All patients received a fixed dose of cisplatin at 80 mg/m2 intravenously (IV) on day 1. The first group received etoposide 50 mg/m2 IV on day 1 and 100 mg orally on days 2 and 3, while all subsequent groups received etoposide 80 mg/m2 IV on day 1 and 160 mg/m2 orally on days 2 and 3. The paclitaxel starting dose was 135 mg/m2 IV over 3 hours and escalated to 175 mg/m2 and 200 mg/m2. Cycles were repeated every 21 days for a maximum of six cycles. Granulocyte colony-stimulating factor was not given prophylactically, but was allowed in subsequent cycles according to American Society of Clinical Oncology guidelines. Nineteen patients were evaluable for toxicity and 18 patients were evaluable for response. Myelosuppression was the major toxicity, with grade 4 neutropenia occurring in 18 of 19 patients (95%), but febrile neutropenia was uncommon and developed in four of 19 patients (21%). Dose-limiting peripheral neuropathy was observed at a paclitaxel dose of 200 mg/m2. Grade 4 nausea/vomiting and diarrhea were also noted at this dose level. Four patients had complete responses (22%) and 13 patients had partial responses (72%). The overall response rate was 94%, with a median survival of 11 months and a 2-year survival rate of 19%. This three-drug combination of paclitaxel with cisplatin and etoposide is highly active with acceptable toxicity. Neurotoxicity was dose limiting at 200 mg/m2 paclitaxel. Neutropenia was frequent but was not associated with significant morbidity. The recommended doses for future clinical trials are paclitaxel 175 mg/m2 IV over 3 hours on day 1 with cisplatin 80 mg/m2 IV on day 1 and etoposide 80/160 mg/m2 IV on day 1 and orally on days 2 and 3. Growth factor support should be used according to American Society of Clinical Oncology guidelines. PMID- 9331139 TI - Paclitaxel and carboplatin in early phase studies: Roswell Park Cancer Institute experience in the subset of patients with lung cancer. AB - The combination of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) given by 3-hour infusion followed by carboplatin infused over 30 minutes has been evaluated in a series of phase I studies and is currently being explored in a phase II study in patients with limited- and extensive-stage small cell lung cancer. Pharmacokinetic measurements were performed at all dose levels in the phase I studies, in which the use of granulocyte colony-stimulating factor in previously treated patients enabled more than twice the dose of paclitaxel to be given with low to moderate doses of carboplatin (dosed to a target area under the concentration-time curve of 4.0 mg x min x mL[-1]). Treatment-naive patients tolerated high paclitaxel doses (270 mg/m2) with carboplatin (dosed to a target area under the curve of 4.5 mg x min x mL[-1]) without granulocyte colony stimulating factor support. Twenty-three patients (including previously treated and untreated) with non-small cell lung cancer were entered at a variety of paclitaxel doses in the phase I studies. At 100 to 205 mg/m2 paclitaxel, none of nine treated patients responded; at 230 to 290 mg/m2, four (29%) of 14 responded. In the phase II study of paclitaxel 250 mg/m2 in previously untreated patients with small cell lung cancer, two of five evaluable patients with extensive-stage disease have shown a partial response. In a preliminary analysis of the pharmacodynamics of paclitaxel in relation to neurotoxicity (dose limiting in two of three phase I studies), neurotoxicity correlated with the total dose of paclitaxel, the area under the curve, and the peak paclitaxel concentration, but not with the length of time plasma paclitaxel levels remained above 0.05 micromol/L. These correlations were not strong, however, and analysis of these data is ongoing. PMID- 9331141 TI - Paclitaxel, carboplatin, and oral etoposide: a phase II trial in limited-stage small cell lung cancer. AB - Carboplatin/etoposide is an active regimen in the treatment of small cell lung cancer. This phase II trial evaluated whether adding paclitaxel (Taxol; Bristol Myers Squibb Company, Princeton, NJ) to this two-drug combination might increase its efficacy. Since April 1996, 55 patients were entered into the ongoing protocol. To date, 35 patients are evaluable for efficacy and toxicity. Most of the evaluable patients are male (28). The patients' median age is 60 years (range, 36 to 74 years); 32 patients have Eastern Cooperative Oncology Group performance status ratings of 1, and the balance are Eastern Cooperative Oncology Group performance status 0. All patients had limited-stage disease. Patients received paclitaxel 175 mg/m2 via 1-hour intravenous infusion on day 1, carboplatin dosed to an area under the concentration-time curve of 5, also on day 1, and oral etoposide 100 mg on days 2 through 8. Overall, 31 patients responded to paclitaxel/carboplatin/etoposide therapy, including complete response in 13 patients (37.1%) and partial response in 18 patients (51.4%). Disease was stable in three patients (8.6%) and disease progressed in one (2.0%). Hematologic toxicity included neutropenia (World Health Organization grade 3 in 24.1% of patients, grade 4 in 31.3%), anemia (4% grade 3, no grade 4), and thrombocytopenia (3.2% grade 3, 2.1% grade 4). Nonhematologic adverse events included minor nausea/vomiting (1.5% grade 3, 9.2% grade 2), polyneuropathy (2.3% grade 2, 17.5% grade 1), and myalgia/arthralgia (8.2% grade 2, 16.4% grade 1). Paclitaxel/carboplatin/etoposide is active in small cell lung cancer with moderate toxicity and good subjective tolerance. There were no life-threatening hematologic or nonhematologic complications in this phase II trial. PMID- 9331142 TI - Defining the role of paclitaxel in lung cancer: summary of recent studies and implications for future directions. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) was first reported to have activity in advanced non-small cell lung cancer (NSCLC) in 1993 and in advanced small cell lung cancer (SCLC) in 1995. Since these original reports, single-agent activity has been confirmed in both NSCLC and SCLC. In NSCLC, the 20% to 25% response rate and median survival times (approximately 40 weeks) are superior to previously reported single-agent therapy. In SCLC, the response rate (> or =50%) and median survival (10 months) are similar to the best previously reported agents. Paclitaxel can be combined safely with both cisplatin and carboplatin. In advanced NSCLC, these two drug combinations produce higher response rates (39% to 42%) than either drug alone. The median survival times reported with the combinations (39 to 45 weeks) are slightly longer than with single-agent paclitaxel. Paclitaxel and cisplatin combinations were shown to be superior to cisplatin and podophyllotoxin combinations in randomized trials. Paclitaxel and paclitaxel plus carboplatin combinations can be safely combined with chest radiotherapy in patients with stage III NSCLC. Response rates and survival times are at least as good as prior best therapies and the results of randomized trials are eagerly awaited. Similarly, paclitaxel and carboplatin combinations produce high response rates when given before surgery for operable patients, and the results of randomized trials are needed to confirm the value of this approach. Paclitaxel-based combinations in advanced SCLC can be administered safely and provide high response rates and relatively long survival times. Randomized trials comparing these combinations to older etoposide/cisplatin combinations are in progress. PMID- 9331145 TI - High seropositivity of anti-CagA antibody in Helicobacter pylori-infected patients irrelevant to peptic ulcers and normal mucosa in Japan. AB - CagA-positive H. pylori is reported to be associated with gastroduodenal disease in Western countries. To evaluate the relationship between CagA and disease, cloning of the entire cagA gene (3771 bp), insertion of a partial fragment (1272 bp) into an expression vector, purification of the recombinant protein, production of an antibody against the recombinant CagA protein through rabbits, and use of the recombinant CagA protein as an antigen, detection of the anti-CagA antibody by western blotting were all performed. Sera of 132 H. pylori-infected patients undergoing endoscopy were studied. Anti-CagA antibodies were detected in 90%, 87%, 90%, 94%, and 93% of patients with gastric ulcer (N = 34), duodenal ulcer (N = 27), chronic gastritis (N = 31), gastric cancer (N = 17), and normal mucosa (N = 15), respectively. High seropositivity of anti-CagA antibody even in individuals with normal mucosa indicated that CagA may not be a unique marker for disease by H. pylori infection in Japan. PMID- 9331144 TI - Clinical application of gastric histology to monitor treatment of dual therapy in H. pylori eradication. AB - This preliminary study attempted to test whether pretreatment gastric histology of H. pylori infection may affect the success of dual therapy and to identify which parameter of gastric histology could be improved after dual therapy. One hundred forty-five dyspeptic patients with H. pylori infection received a two week course of dual therapy (Amoxicillin 500 mg every 6 hr plus omeprazole 20 mg twice a day). In each patient, three pairs of gastric biopsies, sampled from the antrum, lower body, and upper body near the cardia, were collected before treatment and four weeks after completion of dual therapy. The density of H. pylori (score 1-5) and parameters of the modified Sydney system were applied to test the severity of H. pylori-related gastric histology in each specimen. The total bacterial load (score 1-15) was a sum of the density of H. pylori sampled from three biopsies. The overall rate of H. pylori eradication rate by dual therapy is 73.1% (106/145). Univariate analysis of parameters in pretreatment histology disclosed that the presence of mucosal atrophy (P < 0.01), lymphoid follicles (P < 0.005), and higher-density H. pylori (P < 0.001) predisposed to dual therapy failure. Multivariate analysis by stepwise logistic regression further confirmed that both the density of bacteria and the presence of lymphoid follicles are the two major factors related to the outcome of dual therapy (P < 0.001). Four weeks after dual therapy was completed, only patients with successful eradication significantly improved in these gastric histology parameters: acute activity, chronic inflammation, eosinophil infiltration, and mucosal atrophy. However, the lymphoid follicle and intestinal metaplasia were not significantly improved during the study period. The eradication rates among three subgroups with different total bacterial loads (group I: 1-5; II: 6-10; III: 11-15) disclosed a downward trend (I: 89.1%; II: 73%; III: 52.7%). It is concluded that dual therapy could improve gastric histology especially among patients with successful eradication of H. pylori. Evaluating pretreatment histologic parameters, including the density of H. pylori and the presence of lymphoid follicles, is valuable in predicting the success of dual therapy. PMID- 9331146 TI - Cisapride use during human pregnancy: a prospective, controlled multicenter study. AB - The objective of this prospective multicenter study was to determine whether cisapride is associated with increased risk of malformations, spontaneous abortions, or decreased birthweight when used during pregnancy. Cases were paired for age, smoking, and alcohol consumption with controls exposed to nonteratogens, as well as with disease-paired controls. One hundred and twenty-nine pregnant women were exposed to cisapride during pregnancy, including 88 during the period of fetal organogenesis. There were no differences in maternal history, birthweight, gestational age at delivery, and rates of livebirths, spontaneous or therapeutic abortions, fetal distress, and major or minor malformations among groups. It is concluded that exposure to cisapride during pregnancy is not associated with a major increased risk of malformations or spontaneous abortions or with decreased birthweight. PMID- 9331143 TI - Recurrence of Helicobacter pylori infection after successful eradication: nature and possible causes. AB - Recurrence of Helicobacter pylori infection after successful eradication occurs and is associated with relapse of gastroduodenal diseases. The aims of this paper were to assess the incidence and identify the nature and possible causes of recurrence of the infection. A broad-based Medline search was performed to identify all related publications addressing recurrence of the infection between 1986 and 1995. The 12-month recurrence rate varied among the different studies from 0 to 41.5%. A few studies showed 18- to 24-month recurrence rates, which ranged between 0 and 21.4%. Limited data, obtained using molecular fingerprinting techniques, have shown that in most cases recurrence is due to recrudescence of the original strain; a few cases appear to be due to reinfection with a new strain. Recrudescence is most likely during the first 12 months after apparent eradication. Despite the high sensitivity and specificity of the available individual tests for detecting H. pylori infection in untreated patients, no technique alone is sensitive enough to monitor eradication when the four-week rule definition for eradication is used. A combination of two or more techniques increases sensitivity. Sensitivity and specificity are increased when biopsies are taken from both gastric antrum and corpus. The best treatments have the lowest recurrence rates and recurrence is rare when the eradication rate is over 90%. Individual susceptibility and reexposure to H. pylori are suggested as two major causes of reinfection. PMID- 9331147 TI - Increased esophageal chemoreceptor sensitivity to acid in patients after successful reversal of Barrett's esophagus. AB - When compared to patients with erosive esophagitis, patients with Barrett's esophagus have demonstrated reduced chemo- and mechanoreceptor sensitivity to acid infusion and balloon distension, respectively. However, anecdotal clinical experience suggested an increase in symptom perception in patients after successful elimination of Barrett's epithelium, using multipolar electrocoagulation (MPEC) and high-dose proton pump inhibitor (PPI). To determine perception thresholds to acid infusion, we evaluated eight consecutive patients after a series of MPEC treatments resulted in complete elimination of Barrett's mucosa and compared them to 10 age-matched patients with nonreversed Barrett's esophagus and 10 patients with symptomatic, endoscopy-documented erosive esophagitis (Hetzel-Dent grade 2 or greater). Chemosensitivity was determined by a modified acid perfusion test, where acid perception thresholds were quantified by the lag time to initial typical symptom perception, sensory intensity rating, and an acid perfusion sensory score (APSS). While patients after successful elimination of Barrett's esophagus had similar sensory intensity ratings and APSS as patients with erosive esophagitis, the lag times differed significantly between the groups, and both groups had significantly higher sensory intensity ratings and APSS than patients with nonreversed Barrett's esophagus. In conclusion, patients after complete reversal of Barrett's mucosa are unexpectedly as sensitive to acid as symptomatic patients with erosive esophagitis. PMID- 9331148 TI - Ineffective esophageal motility (IEM): the primary finding in patients with nonspecific esophageal motility disorder. AB - Nonspecific esophageal motility disorder (NEMD) is a vague category used to include patients with poorly defined esophageal contraction abnormalities. The criteria include "ineffective" contraction waves, ie, peristaltic waves that are either of low amplitude or are not transmitted. The aim of this study was to identify the prevalence of ineffective esophageal motility (IEM) found during manometry testing and to evaluate esophageal acid exposure and esophageal acid clearance (EAC) in patients with IEM compared to those with other motility findings. We analyzed esophageal manometric tracings from 600 consecutive patients undergoing manometry in our laboratory following a specific protocol from April 1992 through October 1994 to identify the frequency of ineffective contractions and the percentages of other motility abnormalities present in patients meeting criteria for NEMD. Comparison of acid exposure and EAC was made with 150 patients who also had both esophageal manometry and pH-metry over the same time period. Sixty-one of 600 patients (10%) met the diagnostic criteria for NEMD. Sixty of 61 (98%) of these patients had IEM, defined by at least 30% ineffective contractions out of 10 wet swallows. Thirty-five of these patients also underwent ambulatory esophageal pH monitoring. Patients with IEM demonstrated significant increases in both recumbent median percentage of time of pH <4 (4.5%) and median distal EAC (4.2 min/episode) compared to those with normal motility (0.2%, 1 min/episode), diffuse esophageal spasm (0%, 0.6 min/episode), hypertensive LES (0%, 1.8 min/episode), and nutcracker esophagus (0.4% 1.6 min/episode). Recumbent acid exposure in IEM did not differ significantly from that in patients with systemic scleroderma (SSc) for either variable (5.4%, 4.2 min/episode). We propose that IEM is a more appropriate term and should replace NEMD, giving it a more specific manometric identity. IEM patients demonstrate a distinctive recumbent reflux pattern, similar to that seen in patients with SSc. This finding indicates that there is an association between IEM and recumbent GER. Whether IEM is the cause or the effect of increased esophageal acid exposure remains to be determined. PMID- 9331149 TI - The rumination syndrome: clinical and manometric profile, therapy, and long-term outcome. AB - The aims of this study were to investigate the diagnostic studies necessary to identify rumination syndrome and the long-term therapeutic outcomes of patients with rumination syndrome. Sixteen patients with rumination were evaluated between 1989 and 1995. Esophageal motility, gastric emptying, upper gastrointestinal motility, and electrogastrography of all patients were reviewed; follow-up information about their current status was available from 10 of the 16 patients. Duration of symptoms was 77.2 months and the mean age was 28.5 years at the time of diagnosis. Esophageal and upper gastrointestinal motility, gastric emptying, and electrogastrographic studies were all normal. Mean lower esophageal pressure was 12.7 mm Hg and three of the 16 patients had a decreased pressure of less than 6 mm Hg. Ten patients were followed for a mean duration of 31.2 months. Five of 10 patients used biofeedback and relaxation techniques and reported subjective improvement. Our results indicate that rumination syndrome is often confused with a gastric motility disorder and diagnosis is possible if one is aware of this condition. Although there is not a definitive management protocol for this condition, reassurance and education of the patient and the family are crucial first steps followed by behavioral and relaxation programs. PMID- 9331150 TI - Gastric mucosal blood flow regulation in response to different stimuli. AB - We compared changes in gastric mucosal blood flow (GMBF) and left gastric artery blood flow (LGABF) in response to pharmacological, physiological, and pathological stimuli. GMBF and LGABF were measured by the hydrogen gas clearance and perivascular ultrasonic transit time techniques, respectively, under baseline conditions and following intravenous infusion of vasopressin or pentagastrin, isovolemic hemodilution, or gastric perfusion with HCl-taurocholate. Blood flow changes following vasopressin or hemodilution were significantly larger in the left gastric artery than in the gastric mucosa. In contrast, the increment in blood flow associated with pentagastrin-stimulated acid secretion was significantly greater in the gastric mucosa than in the extramural artery. Barrier disruption with acid-taurocholate induced similar changes in both measurement sites. The gastric hyperemia induced by either mechanism was significantly attenuated by blockade of NO synthesis. These data demonstrate that although functional changes in GMBF are primarily supported by changes in blood flow at the extramural gastric arteries, the gastric mucosal microvasculature is also under the influence of independent local control mechanisms. PMID- 9331151 TI - Oxidant-induced activation of nuclear factor-kappa B in cultured guinea pig gastric epithelial cells. AB - The aim of this study was to reveal oxidant-sensitive components in gastric epithelial cells, which may regulate inflammatory processes in gastric mucosa. Gel mobility shift assay showed that treatment of cultured guinea pig gastric epithelial cells with hydrogen peroxide or diamide produced a KB oligonucleotide protein complex within 5 min. The binding proteins consisted of a p50/p65 heterodimer, which was identified by immunosupershift, UV cross-linking, and immunoprecipitation analyses. Immunocytochemical study demonstrated that surface epithelial cells and parietal cells expressed p500 and p65 mainly in the cytosol, and the oxidants rapidly initiated the nuclear translocation of the components. The oxidants caused the up-regulation of p105 (a p50 precursor) synthesis and the expression of inducible nitric oxide synthase mRNA. These results suggest that the oxidant-sensitive p50/p65 heterodimer in gastric epithelial cells may play an important role in transcriptional activation of genes involved in inflammatory responses of the stomach. PMID- 9331152 TI - Protection against chemically-induced oxidative gastrointestinal tissue injury in rats by bismuth salts. AB - Oxygen free radicals (OFR) are implicated in the pathogenesis of stress, chemically induced gastric lesions, and gastrointestinal injury. The concentration-dependent scavenging abilities of bismuth subsalicylate (SBS), colloidal bismuth subcitrate (CBS), and selected OFR scavengers, including superoxide dismutase (SOD), catalase, mannitol, and allopurinol were examined against biochemically or chemically generated superoxide anion, hydroxyl radical, and hypochlorite radical plus hypochlorous acid based on a chemiluminescence assay. Furthermore, both gastric (GM) and intestinal mucosa (IM) were individually exposed in vitro to these free radical generating systems, and the concentration-dependent protective abilities of SBS and CBS against lipid peroxidation (LP) were compared with selected OFR scavengers. In addition, 24-hr fasted rats were orally treated with the necrotizing agents 0.6 M HCl, 0.2 M NaOH, 80% ethanol, and aspirin (200 mg/kg). The extent of tissue injury in the GM and IM was determined by assessing LP, DNA fragmentation, and membrane microviscosity. Dose- and time-dependent in vivo protective abilities of CBS (100 mg/kg) and SBS (15 mg/kg) were also assessed. Following incubations with superoxide anion and hydroxyl radical generating systems in the presence of 125 mg SBS/liter, approximately 47% and 61% inhibitions were observed in the chemiluminescence response, respectively, while 48% and 46% inhibitions were observed with 125 mg CBS/liter. SBS and CBS exerted similar abilities towards hypochlorite radical plus hypochlorous acid. Approx. 3.1- and 3.7-fold increases in LP were observed in the GM and IM of rats following oral administration of 0.6 M HCl. Pretreatment of the rats with SBS and CBS decreased 0.6 M HCl-induced LP in the GM by approx. 39% and 27%, respectively, with similar decreases in LP in the IM. SBS exhibited better protective abilities towards 0.6 M HCl and 0.2 m NaOH-induced GM and IM injury as compared to CBS. SBS and CBS provided similar protection towards 80% ethanol-induced gastric injury, while CBS exerted a superior protective ability towards aspirin-induced gastric injury. The results demonstrate that both SBS and CBS can scavenge reactive oxygen species and prevent tissue damage produced by OFR. PMID- 9331153 TI - Role of gastrin/CCK-B receptor in the regulation of gastric acid secretion in rat. AB - The aim of this study was to investigate whether gastrin regulates morphological changes and alpha-subunit gene expression in parietal cells through the gastrin/CCK-B receptor on enterochromaffin-like cells by histamine release. Treatment with 100 mg/kg of YM022, a potent and selective gastrin/CCK-B receptor antagonist, for one week in rats did not alter mRNA levels of histidine decarboxylase or H+, K+-ATPase. However, parietal cell morphology predominantly changed to the resting form, although the serum gastrin concentration was significantly increased. Additional treatment with YM022 and oral omeprazole, 100 mg/kg, for one week markedly suppressed the increases of mRNA levels of histidine decarboxylase and H+, K+-ATPase and completely blocked the morphological transformation of the parietal cells to a stimulated form induced by treatment with omeprazole alone. This indicates that the morphological transformation of parietal cells to an activated form with a subsequent increase in H+, K+-ATPase synthesis caused by hypergastrinemia is mediated by increased histidine decarboxylase gene expression in enterochromaffin-like cells via gastrin/CCK-B receptors. PMID- 9331154 TI - ACE inhibition by enalaprilate stimulates duodenal mucosal alkaline secretion via a bradykinin pathway in the rat. AB - The effects of enalaprilate on duodenal mucosal alkaline secretion (in situ titration) and mean arterial blood pressure were investigated in chloralose anesthetized male rats. A bolus injection of enalaprilate (0.7 mg/kg intravenously) increased alkaline secretion by about 60%, and this response was resistant to guanethidine (5 mg/kg intravenously), splanchnicotomy, and vagotomy. Furthermore, angiotensin II infusion (0.25-2.5 microg/kg/hr intravenously) following the administration of enalaprilate failed to influence this response. Bradykinin (10(-6)-10(-4) M) applied topically to the serosal surface of the duodenal segment under study increased dose-dependently the duodenal mucosal alkaline secretion, an effect that could be blocked by the selective bradykinin receptor subtype-2 antagonist HOE140 (100 nmol/kg intravenously). HOE140 also antagonized the response to enalaprilate. These data suggest that enalaprilate increases duodenal mucosal alkaline secretion via a local bradykinin pathway involving receptors of the bradykinin receptor subtype-2 antagonist, rather than by blockade of endogenous angiotensin II or by central autonomic neural regulation. PMID- 9331155 TI - Esophageal white sponge nevus associated with severe dysphagia and odynophagia. PMID- 9331157 TI - Neurofilament and intermediate filament immunoreactivity in human intestinal myenteric neurons. AB - It has been reported previously that rat myenteric neurons have neurofilament (NF) immunoreactivity that differs from the brain. Now the result of a study of neurofilaments and intermediate filament immunoreactivity in human colon and ileum using a panel of antibodies and indirect immunofluorescence techniques is reported here. Results with polyclonal neurofilament antisera showed positive immunoreactivity in subsets of myenteric neurons. Results with peripherin and alpha-internexin showed immunoreactivity in some neurons that contained neurofilaments and in many that were neurofilament negative, similar to our observations in rat. Some monoclonal antibodies to epitopes on NF-M and NF-H demonstrated weak or negative immunoreactivity in human myenteric neurons yet showed positive immunoreactivity in brain. Some of these antibodies are phosphorylation dependent, suggesting NF-M and NF-H epitopes in myenteric neurons are not as phosphorylated as in brain; other antibodies are phosphorylation independent, suggesting other differences or masking of epitopes. In summary, neurofilaments are present in a subset of myenteric neurons. In those human myenteric neurons that contain them, the neurofilaments appear immunologically distinct from those in the brain. PMID- 9331156 TI - Electric field stimulation-induced guinea pig gallbladder contractions: role of calcium channels in acetylcholine release. AB - Gallbladder motility is modulated by intrinsic cholinergic neurons. The aims of this study were to determine: (1) the effect of electric field stimulation (EFS) on guinea pig gallbladder smooth muscle, and (2) the role of calcium channels in mediating neurotransmitter release. Gallbladder muscle strips were studied isometrically in vitro. EFS (1-16 Hz, 100 V, 0.5-msec pulse width, 30-sec train duration) was used to activate the intrinsic nerves. Exogenous acetylcholine was also used to directly stimulate the smooth muscle. EFS produced a frequency dependent contractile response that was completely abolished by tetrodotoxin. EFS induced contractions at 16 Hz were suppressed by 84 +/- 4% with atropine, whereas hexamethonium had no effect. The L-type calcium channel blocker, nifedipine, reduced EFS contractions by 51 +/- 4%, whereas it reduced contractions to acetylcholine by only 11 +/- 5%. The N-type calcium channel blocker, omega conotoxin GVIA, reduced EFS-induced contractions by 22 +/- 9%, but did not affect acetylcholine-induced contractions. EFS-induced contractions of the guinea pig gallbladder are primarily mediated by activation of postganglionic cholinergic neurons. The acetylcholine release from these cholinergic neurons is regulated by L- and N-type calcium channels. The inhibitory effect of calcium channel blockers on the gallbladder seen in vivo may be in part related to inhibition of acetylcholine release from the intrinsic cholinergic nerves of the gallbladder. PMID- 9331158 TI - Effects of medium-chain and long-chain triglycerides on antroduodenal motility and small bowel transit time in man. AB - Medium-chain triglycerides are known to induce diarrhea, possibly resulting from accelerated intestinal transit. We performed antroduodenal manometry and lactulose hydrogen breath testing simultaneously in eight healthy subjects in order to determine the effects of intraduodenally administered medium-chain triglycerides (MCT) and long-chain triglycerides (LCT) on gastrointestinal motility and small bowel transit time. LCT (15 mmol/hr) induced a fed motor pattern. In contrast, during MCT, in both equimolar (15 mmol/hr; MCT-1) and equicaloric (30 mmol/hr; MCT-2) amounts comparable to LCT, interdigestive motility was preserved but with a significantly (P < 0.05) shorter MMC cycle length (MCT-1, 65 +/- 7 min; MCT-2, 53 +/- 6 min) compared to control (saline infusion; 127 +/- 14 min). Duodenocecal transit time (DCTT) was significantly (P < 0.05) accelerated during administration of MCT (MCT-1, 56 +/- 6 min; MCT-2, 69 +/- 9 min) and was not affected by LCT (105 +/- 13 min) when compared to control (101 +/- 9 min). IN CONCLUSION: MCT, in contrast to LCT, preserve interdigestive motility with a shorter MMC cycle length and accelerate DCTT. PMID- 9331159 TI - Tachykinins influence interdigestive rhythm and contractile strength of human small intestine. AB - The effect of the putative enteric neurotransmitters neurokinin A and substance P were investigated on human small intestinal motility. Either neurokinin A, at doses of 6-25 pmol/kg/min, or substance P at doses of 1-6 pmol/kg/min were administered intravenously to healthy volunteers over 4 hr. Neurokinin A dose dependently increased the fraction of phase II of the migrating motor complex, contraction frequency, motility index, and amplitude of contractions. At the highest dose, neurokinin A induced a phase II-like pattern, disrupting the migrating myoelectric complex. Substance P dose-dependently increased phase II of the migrating motor complex. The contraction frequency increased slightly at the highest dose, but neither motility index nor contraction amplitude changed. It is concluded that neurokinin A and substance P stimulate small intestinal motility in man, and it can be speculated that they play a role in the control of human small intestinal motility. PMID- 9331161 TI - Barostat review. PMID- 9331160 TI - Change in colonic motility after extrinsic autonomic denervation in dogs. AB - Changes in colonic motility were compared in dogs undergoing autonomic denervation of the paraaortic and presacral (group A), paraaortic (group B), or mesocolonic region (group C), and sham operation (group D). Five bipolar recording electrodes were placed into the seromuscular layer of the colon and rectum. The numbers of continuous electrical response activity and contractile electrical complex after an intragastric olive oil injection were smaller in group A than in the other groups (P < 0.05) from three weeks through six months after denervation. This difference was significant even in the proximal colon. These data suggest that the pelvic plexus may play an important role in colonic motility including the proximal colon. The damage to the plexus did not recover for at least six months after denevation. Pelvic plexus injury may thus be one of possible explanations for the prolonged change in bowel habit after anterior resection of the rectum. PMID- 9331163 TI - Evidence for a nonneural electrogenic effect of cholera toxin on human isolated ileal mucosa. AB - Cholera toxin-induced intestinal secretion in intact rats requires a functioning myenteric plexus. The aim of this investigation was to determine whether neural elements were essential for cholera toxin to produce a secretory effect in human isolated ileum. Mucosal preparations were mounted in Ussing chambers. Cholera toxin was applied apically and short-circuit current monitored for 3 hr, at which point forskolin was given. Cholera toxin (10 microg/ml) induced a tetrodotoxin insensitive increase in short-circuit current in muscle-stripped preparations of human ileum. The increase was not additive with the action of forskolin (25 microM). Cholera toxin exerts a marked nonneural secretory effect in human ileal mucosa in vitro, probably by the same mechanism as forskolin, namely elevation of cyclic AMP. PMID- 9331162 TI - Colchicine is an effective treatment for patients with chronic constipation: an open-label trial. AB - Chronic constipation is a common clinical condition that frequently does not respond to routine therapeutic measures. We hypothesized that colchicine would be effective in this condition because we reported that it stimulates intestinal motility in rats and commonly causes diarrhea in patients taking the drug for either gouty arthritis or Familial Mediterranean fever. We prospectively studied seven patients with chronic constipation who were refractory to medical therapy and treated them with oral colchicine 0.6 mg per os three times a day for eight weeks in an open-label pilot study. During the study, the mean number of spontaneous bowel movements significantly increased (P < 0.05) from 1.7 +/- 0.5 noted during routine therapy of constipation with laxatives and enemas to 6.4 +/- 0.7 per week; mean colonic transit time significantly (P < 0.05) decreased from 58.1 +/- 2.5 to 47.1 +/- 5.0 hr; and symptoms of abdominal pain, nausea, and bloating significantly (P < 0.05) improved during therapy with colchicine. Oral colchicine (0.6 mg three times a day) therapy appears to be an a promising treatment for chronic constipation and a placebo-controlled trial is indicated to confirm these findings. PMID- 9331164 TI - Protease activity in a hapten-induced model of ulcerative colitis in rats. AB - Inflammatory bowel disease (IBD) is a painful and debilitating condition affecting the mucosal lining of the colon and other areas of the gastrointestinal tract. IBD generally falls into two major categories: ulcerative colitis (UC) and Crohn's disease. We have utilized dinitrobenzenesulfonic acid (DNBS) to induce experimental UC in rats. Histopathologic analysis indicates that DNBS induces a condition in animals similar to human UC. Biochemical results revealed 6- to 10 fold elevated levels of serine protease activity in colon tissue from animals with UC as compared with matched controls. We also observed elevated levels of protease activity in tissue samples obtained from human patients with UC. Hence, our results demonstrate that protease activity is increased in rodent and human UC. These proteases may play a significant role in destruction of colonic tissue in IBD. Protease inhibitors that target serine proteases may be useful pharmacological agents to limit tissue destruction in IBD. PMID- 9331166 TI - Induction of a 72-kDa heat shock protein and cytoprotection against thioacetamide induced liver injury in rats. AB - Heat shock proteins are ubiquitous intracellular proteins induced by various physiological stress-related events. A 72-kDa heat shock protein (HSP72) has been reported to be an endogenous cytoprotectant in variety of cells in vitro. In order to study the cytoprotective function of HSP72 in the liver, the effect of preinduction of HSP72 in rat liver by systemic hyperthermia on thioacetamide induced hepatic injury was investigated in this study. Expression of HSP72 in the liver was investigated by immunoblot and densitometric analysis. Rats were injected with thioacetamide (100 mg/kg, subcutaneously) with or without preinduction of HSP72 by hyperthermia. Serum AST and ALT concentrations were measured before and after thioacetamide injection in both group. Histologic alteration of the liver was evaluated also. Systemic hyperthermia (42.5 degrees C, 20 min) significantly induced HSP72 in the liver. Thioacetamide-induced hepatic injury was clearly prevented by preinduction of HSP72 by hyperthermia. Prevention of hepatocyte damage was more clear in the area around central veins where HSP72 induction was apparent. Our findings might suggest that HSP72 has an important function in the liver with respect to cytoprotection. These results might be important for understanding the mechanism of "adaptive cytoprotection" in the liver mediated by the function of heat shock proteins as "molecular chaperons" as reported in vitro. PMID- 9331165 TI - Effect of interferon-alpha2b administration on rat liver regeneration after partial hepatectomy. AB - The purpose of the present study was to delineate the effect of interferon alpha2b (IFN-alpha2b) administration on the liver regenerative capacity after partial hepatectomy in rats. The administration of IFN-alpha2b simultaneously with partial hepatectomy did not affect hepatic proliferation in a statistically significant manner. When IFN-alpha2b was administered either 2 or 12 hr postoperatively, an inhibition of hepatocyte proliferation was observed 24 hr postoperatively, while at further time intervals up to 48 hr, DNA synthesis remained similar to that observed in the simply partially hepatectomized rats. The enzyme thymidine kinase (TK), has been implicated in the suppression of proliferation in interferon-treated cell cultures. In all IFN-alpha2b-treated groups of rats, alterations of TK activity were observed without being correlated to the liver regenerative status. Additionally, the administration of the polyamine putrescine in partially hepatectomized rats treated at the time of surgery with IFN strongly enhanced TK activity, but did not affect DNA biosynthesis. In the above-mentioned in vivo model of controlled cellular proliferation, the administration of IFN-alpha2b affected the rate of hepatocyte proliferation depending on the time of its administration; this effect was not correlated to the enzymatic activity of TK, as inhibited TK activity is responsible for the suppressed DNA synthesis in in vitro systems. PMID- 9331168 TI - Activity-dependent regulation of calcium-binding proteins in the developing rat olfactory bulb. AB - Intracellular calcium, important in a variety of second messenger cascades, is regulated in part by calcium-binding proteins such as calretinin, parvalbumin, and calbindin. These proteins are highly concentrated in the rat main olfactory bulb and are localized in distinct neuronal populations. In the present study, postnatal expression was characterized immunohistochemically in normal rats and in rats with functional olfactory deprivation caused by unilateral naris closure, a manipulation that attenuates electrical activity in the bulb. Bulbs were examined from rats that had undergone naris closure or sham surgery on either postnatal day 1 (P1) or P30 and were allowed varying subsequent survival times. Each of the calcium-binding proteins showed both distinct patterns of early expression and differential susceptibility to olfactory restriction. For example, at P10, the densest immunoreactivity was observed for calretinin, a protein whose expression was the least affected by naris closure. After occlusion from P1-P30, there was a 30% reduction in the density of calbindin-immunoreactive profiles in the glomerular layer, and parvalbumin-immunoreactive profiles were reduced by 64% in the external plexiform layer. Unlike many other changes induced by deprivation, the effects of olfactory restriction on calbindin and parvalbumin expression were not age dependent: naris closure from P30-P60 caused similar substantial decreases in calbindin and parvalbumin immunoreactivities. These data demonstrate that the expression of calbindin and parvalbumin in rat bulb is regulated, in part, by afferent activity that is associated with full sensory experiences. The reductions of these calcium-binding proteins following olfactory deprivation are likely to be commensurate with altered control of intracellular calcium. PMID- 9331167 TI - Carrier mediated delivery of NGF: alterations in basal forebrain neurons in aged rats revealed using antibodies against low and high affinity NGF receptors. AB - The distribution of low and high affinity nerve growth factor (NGF) receptors was investigated in the basal forebrain during aging and NGF treatment. A peripheral administration model for NGF was utilized. NGF was conjugated to a transferrin receptor antibody (OX-26-NGF), and this conjugate was injected into the tail vein of aged Fischer 344 male rats (24 months) twice weekly for 5 weeks (equivalent to 50 microg of NGF/injection). Controls were injected with a non-conjugated mixture of OX-26 and NGF. The aged rats treated with conjugate showed a significant increase in cell size of p75- and trkA-immunoreactive neurons in the medial septal nucleus and vertical limb of the diagonal band as compared to controls. A significant increase in cell size of trkA-immunoreactive neurons was also observed in the horizontal limb of the diagonal band in rats treated with conjugate. Rats treated with conjugate also showed a significant increase in overall staining density for p75 and trkA antibodies in the medial septal nucleus as compared to controls. A significant increase in staining density of p75 immunoreactive structures was also observed in the vertical and horizontal limbs of the diagonal band. Therefore, treatment with OX-26-NGF conjugate has regional effects on both the low and high affinity NGF receptors in terms of cell body size and staining density in the basal forebrain of aged rats. The current findings support the idea that this delivery system might be useful in therapeutic approaches involving the delivery of neurotrophic factors and other large molecules into the brain. PMID- 9331169 TI - Colchicine affects cortical and amygdalar neurochemical changes differentially after middle cerebral artery occlusion in rats. AB - Recently, we have shown increases in the immunoreactivity for neuropeptide Y and tyrosine hydroxylase in the insular cortex surrounding the focal infarction after middle cerebral artery occlusion. In addition, the immunoreactivity for neuropeptide Y, leucine-enkephalin, dynorphin, and neurotensin increased ipsilaterally in the amygdala. Increases in immunoreactivity were observed in nerve terminals and fibers; changes in the neuropeptides were maximal 3 days after stroke. Local excitotoxic injury of the insular cortex also elicited similar neuropeptide changes unilaterally in the same regions. In this study, immunohistochemistry was used following intracerebroventricular injection of colchicine and stroke to determine whether blockade of axonal transport would prevent these neurochemical changes. These experiments would also locate the putative cellular origins of the neurochemicals involved. Control rats received either colchicine injection or middle cerebral artery occlusion alone. Injection of colchicine enhanced the periinfarct increase in neuropeptide Y but did not alter the increase in tyrosine hydroxylase. The neuropeptide Y increase was observed in local cortical neurons. Colchicine prevented the increases in immunoreactivity for the neuropeptides in the amygdala on the side of stroke, although there were small perikarya that showed immunoreactivity for these neuropeptides within the amygdala on both sides. We conclude that local cortical neurons are responsible for the increase in neuropeptide Y in the periinfarct region, that the cortical increase in tyrosine hydroxylase is not dependent on fast axonal transport, and that axonal transport of signals from the insular cortex to the amygdala is critical in mediating the amygdalar neuropeptide changes seen after stroke. PMID- 9331170 TI - Three types of serotonin-containing amacrine cells in tadpole retina have distinct clonal origins. AB - In the Xenopus tadpole there are three different serotonin-containing amacrine cells: large, brightly fluorescent (LB), and small, dimly fluorescent (SD) cells in the inner nuclear layer and displaced (DIS) cells in the ganglion cell layer. To reveal the potential roles of regional cues and lineage factors in their determination, quantitative maps were made of the spatial distribution and blastomere origin of each cell type. LB and SD cells were evenly distributed across the four retinal quadrants, arguing against a hypothesis that these cells are induced differentially by quadrant-specific cues. Blastomere progenitors of the 32-cell embryo are biased to produce only subsets of serotonin amacrine cells: 1) all nine progenitors of one retina produced some SD cells, but only eight produced LB, and only five produced DIS cells; and 2) there are overlapping but distinct subsets of blastomere progenitors for each serotonin subtype. This bias is not simply a reflection of the size of a clone in the retina; significant quantitative differences were observed between the proportion of serotonin progeny and the proportion of the entire retina produced by six of the nine retinal progenitors. This bias also is not simply a reflection of the spatial distribution of a blastomere clone in the retina; the number of LB descendants in each retinal quadrant was statistically different from its progenitor's total contribution to the quadrant. These results indicate that the development of the three different serotonin-containing amacrine cells in the retina is biased by membership in specific blastomere clones. PMID- 9331171 TI - Molecular cloning of a cDNA encoding the neuropeptides APGWamide and cerebral peptide 1: localization of APGWamide-like immunoreactivity in the central nervous system and male reproductive organs of Aplysia. AB - While much is known about the neural and endocrine mechanisms that control egg laying in the gastropod mollusk Aplysia, relatively little is known about the regulation of male reproductive activity in this simultaneous hermaphrodite. In the present study, we have cloned and sequenced a cDNA that encodes a precursor protein, the predicted posttranslational processing of which presumably generates nine copies of the neuropeptide Ala-Pro-Gly-Trp-NH2 (APGWamide), five connecting peptide sequences, and a C-terminal peptide. The sequence of one connecting peptide is identical to the previously characterized cerebral peptide 1. Northern blot analysis identified two major APGWamide mRNA transcripts (approximately 1.3 kb, approximately 2.4 kb), which were present in central nervous system ganglia, but were most abundant in the right cerebral and right pedal ganglia. Immunohistochemical studies using sexually mature Aplysia demonstrated that the vast majority of APGWamide-like immunoreactivity was localized in 30-40 neurons along the anterior and medial margins of the right cerebral ganglion and in a cluster of 15-20 neurons in the right pedal ganglion. A total of only about ten immunoreactive neurons were located in other ganglia. Immunohistochemistry also demonstrated that APGWamide was present in the reproductive organs that participate in the storage or transport of sperm, including the small hermaphroditic duct (site of sperm storage before mating), the white hemiduct (also known as the copulatory duct), and penial complex. As a group, these data suggest that APGWamide may play a role in regulating male reproductive function in Aplysia, as it does in other gastropods. PMID- 9331172 TI - Patterns of connectivity in a Drosophila nerve. AB - We investigated the spatial patterns of synaptic profiles in en passant synapses between the premotor axon of a peripherally synapsing interneuron (PAPSI) and a set of individually identifiable motoneuron axons in Drosophila melanogaster. These synaptic profiles are distributed as the axons travel parallel to each other in a bundle; the synapses begin as the axons leave the thoracic ganglion and continue peripherally for 45-65 microm. We found that the number of synaptic profiles per micron length of the motoneuron axons was greatest close to the ganglion; the cumulative distribution of profiles could be fitted to curves of the form f(x) = alpha(1 - e(-beta x)), where x = the distance from the thoracic ganglion, and alpha and beta are constants. The distribution of synaptic profiles was also examined in a mutant strain, Passover (Pas), known to affect connectivity in a pathway that includes the PAPSI. The synaptic profiles between the PAPSI and the motoneuron axons appeared ultrastructurally unremarkable in Pas. Also, the total number of synaptic profiles between the PAPSI and the motoneuron axons did not differ between Pas and wild type flies. However, the distribution of synaptic profiles among the individual motoneuron axons did differ significantly from wild type flies, as did the area of contiguity between the motoneuron axons and the PAPSI, which was much greater in Pas than in wild type flies. PMID- 9331173 TI - Effects of excitatory amino acids on neuromuscular development in the chick embryo. AB - To investigate the presumptive role of excitatory amino acids (EAAs) in the regulation of normally occurring motoneuron (MN) death, chick embryos were treated with the glutamate receptor antagonists dizocilpine maleate and 1,2,3,4 tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium. Both failed to alter the number of surviving MNs at the end of the critical period of programmed cell death. However, treatment with 3-(2-carboxypiperazin-4-yl)-propyl 1-phosphonic acid, a competitive N-methyl-D-aspartic acid (NMDA) receptor antagonist, was able to rescue a significant number of MNs from death. Treatment with several EAA agonists induced extensive excitotoxic lesions in the spinal cord. MN degeneration induced by excitotoxins exhibited changes characteristic of necrosis rather than apoptosis. However, when either 0.5 or 1 mg of NMDA was applied acutely on embryonic day (E) 7, about 50% of treated embryos failed to exhibit NMDA-induced excitoxicity but rather showed a clear reduction in the number of pyknotic MNs. This apparent neuroprotective effect of NMDA was also observed in a subset of embryos chronically treated with NMDA, in which an excessive number of MNs was detected when examined on E9. Surprisingly, in the same experiment other embryos showed either normal or highly reduced MN numbers. Embryos with motoneuronal depletion induced by NMDA also showed a delayed impairment of later neuromuscular development with the appearance of degenerative changes in surviving MNs and apoptosis of skeletal muscle cells. Because some of the alterations reported here are similar to those described in MN diseases, our experimental model may be useful for gaining insights into the mechanisms that control both developmentally regulated and pathological MN death. PMID- 9331174 TI - Grafts of EGF-responsive neural stem cells derived from GFAP-hNGF transgenic mice: trophic and tropic effects in a rodent model of Huntington's disease. AB - The present study examined whether implants of epidermal growth factor (EGF) responsive stems cells derived from transgenic mice in which the glial fibrillary acid protein (GFAP) promoter directs the expression of human nerve growth factor (hNGF) could prevent the degeneration of striatal neurons in a rodent model of Huntington's disease (HD). Rats received intrastriatal transplants of GFAP-hNGF stem cells or control stem cells followed 9 days later by an intrastriatal injection of quinolinic acid (QA). Nissl stains revealed large striatal lesions in rats receiving control grafts, which, on average, encompassed 12.78 mm3. The size of the lesion was significantly reduced (1.92 mm3) in rats receiving lesions and GFAP-hNGF transplants. Rats receiving QA lesions and GFAP-hNGF-secreting grafts stem cell grafts displayed a sparing of striatal neurons immunoreactive (ir) for glutamic acid decarboxylase, choline acetyltransferase, and neurons histochemically positive for nicotinamide adenosine diphosphate. Intrastriatal GFAP-hNGF-secreting implants also induced a robust sprouting of cholinergic fibers from subjacent basal forebrain neurons. The lesioned striatum in control grafted animals displayed numerous p75 neurotrophin-ir (p75NTR) astrocytes, which enveloped host vasculature. In rats receiving GFAP-hNGF-secreting stem cell grafts, the astroglial staining pattern was absent. By using a mouse-specific probe, stem cells were identified in all animals. These data indicate that cellular delivery of hNGF by genetic modification of stem cells can prevent the degeneration of vulnerable striatal neural populations, including those destined to die in a rodent model of HD, and supports the emerging concept that this technology may be a valuable therapeutic strategy for patients suffering from this disease. PMID- 9331175 TI - Subnuclear localization of FOS-like immunoreactivity in the parabrachial nucleus after orofacial nociceptive stimulation of the awake rat. AB - We used FOS-like immunohistochemistry to detect neuronal activity in the pontine parabrachial nucleus after injection of formalin into the lower lip of the awake rat and compared the labeling pattern with that seen after formalin injection into the hindpaw. One hour after a formalin injection into the lip, many FOS immunoreactive cells were seen in the parabrachial nucleus, preferentially on the side ipsilateral to the injection site. Detailed anatomical analysis revealed that FOS-immunoreactive neurons were localized predominantly to three regions of the parabrachial nucleus: the external lateral, the external medial, and the Kolliker-Fuse subnuclei, with sparser labeling present in the dorsal and superior lateral subnuclei and in the medial parabrachial nucleus. In contrast, a formalin injection into the hindpaw resulted in dense FOS-labeling in the superior, dorsal, and central lateral subnuclei, with sparse to moderate labeling in the Kolliker-Fuse nucleus, and sparse labeling in the external lateral and external medial subnuclei, as described previously (Hermanson and Blomqvist, J. Comp. Neurol., [1996] 368:45-56). The distribution of FOS-labeled neurons after noxious orofacial stimulation corresponds to the termination pattern in the parabrachial nucleus of fibers that originate from neurons in the marginal zone of the trigeminal dorsal horn and is different from that seen after nociceptive stimulation of other body parts. Considering the differences in efferent connections of parabrachial subnuclei, the present findings imply that noxious information from the orofacial region to the parabrachial nucleus has other functional roles than noxious information from the trunk and limbs. Such roles may include the integration of somatosensory and gustatory information, which has been suggested to be of importance for feeding behavior. PMID- 9331176 TI - Hypothalamic neurohistochemistry of the murine anorexia (anx/anx) mutation: altered processing of neuropeptide Y in the arcuate nucleus. AB - Neuropeptide Y is one of the most powerful neurochemical stimulants of food intake known. The neuronal substrate for this action is believed to be the neuropeptide Y-expressing cell population in the hypothalamic arcuate nucleus. In this study, mice homozygous for the anorexia mutation (anx) were investigated histochemically; anx is a recessive mutation that causes decreased food intake and starvation, leading to death 22 days after birth. We were interested to see whether any hypothalamic neurochemical abnormalities could be detected in this genetic model of starvation. By using immunohistochemistry and in situ hybridization, the hypothalamic distributions of neuropeptide Y, cholecystokinin, galanin, and serotonin, all messenger molecules postulated to be involved in the regulation of food intake and energy metabolism, were investigated. Immunoreactivities for somatostatin, the excitatory amino acid aspartate, and acetylcholinesterase were also studied. Neuropeptide Y-like immunoreactivity was increased markedly in arcuate cell bodies and decreased in terminals in the arcuate nucleus and other hypothalamic regions of anx/anx mice compared with normal litter mates. In situ hybridization for neuropeptide Y mRNA, however, showed no significant difference in gene expression in the arcuate nucleus. In addition, immunoreactivities for aspartate, acetylcholinesterase, and somatostatin in the arcuate nucleus were decreased in anx/anx mice. For cholecystokinin, galanin, and serotonin, no certain differences in hypothalamic immunoreactivity could be seen. These data suggest that a defect in neuropeptide Y-ergic signalling in the arcuate neurons may contribute to the failure to thrive in anx/anx mice. PMID- 9331178 TI - Evidence from normal and degenerating photoreceptors that two outer segment integral membrane proteins have separate transport pathways. AB - Detachment of the neural retina from the retinal pigment epithelium induces photoreceptor degeneration. We studied the effects of this degeneration on the localization of two photoreceptor outer segment-specific integral membrane proteins, opsin and peripherin/rds, in rod photoreceptors. Results from laser scanning confocal microscopic and electron microscopic immunolocalization demonstrate that these two proteins, normally targeted to the newly-forming discs of the outer segments, accumulate in different sub-cellular compartments during photoreceptor degeneration: opsin immunolabeling increases throughout the photoreceptor cell's plasma membrane, while peripherin/rds immunolabeling occurs within cytoplasmic vesicles. The simplest hypothesis to explain our results is that these proteins are transported in different post-Golgi transport vesicles and separately inserted into the plasma membrane. More complex mechanisms involve having the two co-transported and then opsin finds its way into the plasma membrane but peripherin/rds does not, remaining behind in vesicles. Alternatively, both insert into the plasma membrane but peripherin/rds is recycled into cytoplasmic vesicles. We believe the data most strongly supports the first possibility. Although the transport pathways for these proteins have not been fully characterized, the presence of peripherin/rds-positive vesicles adjacent to the striated rootlet suggests a transport role for this cytoskeletal element. The accumulation of these proteins in photoreceptors with degenerated outer segments may also indicate that their rate of synthesis has exceeded the combined rates of their incorporation into newly forming outer segment disc membranes and their degradation. The accumulation may also provide a mechanism for rapid recovery of the outer segment following retinal reattachment and return of the photoreceptor cell to an environment favorable to outer segment regeneration. PMID- 9331177 TI - Morphology and electrophysiology of dentate granule cells in the rhesus monkey: comparison with the rat. AB - The morphologic and electrophysiologic properties of dentate granule cells in the young adult rhesus monkey (Macaca mulatta) were examined for the first time with whole-cell patch clamp recordings and intracellular biocytin filling in in vitro hippocampal slice preparations. Data from monkeys were compared with data generated in an identical manner from adult Sprague-Dawley rats. Intracellularly filled monkey and rat granule cells were identical in numerous morphologic parameters, including area of somata, total dendritic length, dendritic spread, segment number and length, and branching pattern. The single statistically significant difference in morphology was the vertical extent of the dendritic tree (distance from soma to fissure), which was 20% greater in the monkey. The passive membrane properties (resting membrane potential, input resistance, and membrane time constant) measured under current clamp conditions were virtually identical. The thresholds and amplitudes of action potentials were the same, but significant differences were seen in the kinetics of single action potentials. Monkey granule cell action potentials were significantly longer in duration (with slower rise and fall times) than action potentials in rat cells. These differences were likely due to a much smaller fast after hyperpolarization in the monkey as compared with the rat cells. Thus, with the exception of action potential properties, the principal finding of this study is that there is significant conservation of both form and function in dentate granule cells in these two species, despite the enormous phylogenetic separation. This suggests that granule cell properties may be extremely stable across diverse mammalian species. PMID- 9331180 TI - Economic risks of refractive surgical procedures. PMID- 9331179 TI - Neurons of the Drosophila giant fiber system: I. Dorsal longitudinal motor neurons. AB - The giant fiber system (GFS) mediates the startle response of Drosophila. This response includes an activation of the dorsal longitudinal wing-depressor muscles (DLMs). However, the morphology of the motor neurons innervating these muscles has not been well studied. Even the location of the somata of these motor neurons has been a source of controversy. This paper identifies the somata and provides a morphological description of these motoneurons. The DLM is comprised of six muscle fibers, named a through f (dorsal to ventral). Each muscle fiber is singly innervated. Each of the four ventral muscle fibers is innervated by a separate motor neuron (DLMn c-f), but the two dorsal fibers share an axon (DLMn a/b). Motor neurons were back filled by introducing horseradish peroxidase (HRP) into individual muscle fibers. The cell body of DLMn a/b is extraordinarily large (32 microm) and lies dorsal and contralateral. In this hemiganglion, it does not have a fixed position; it can be found anywhere from the midline to the extreme lateral edge of the ganglion. The position is not genetically controlled: We find no strain differences, and, within a single individual, the right and left cells may take different positions. The neuritic arborization fills a shallow dorsal cap of the ganglion, with branches arrayed like a feather. The cell bodies of the four motor neurons c-f lie in an ipsilateral and ventral cluster. Each soma occupies a fixed corner of this quadrilaterally shaped cluster. The neurites ramify in the same dorsal region as DLMn a/b. PMID- 9331181 TI - Reduction of IOP after PRK. PMID- 9331182 TI - Reduction of IOP after PRK. PMID- 9331183 TI - 5-Fluorouracil in primary combined glaucoma surgery. PMID- 9331184 TI - Sports-related ocular injuries. PMID- 9331185 TI - Does warming of anesthetic solutions improve analgesia and akinesia in retrobulbar anesthesia? PMID- 9331186 TI - Reproducibility of nerve fiber layer thickness measurements. PMID- 9331187 TI - Immunosuppression and HIV in young patients with ocular surface dysplasia. PMID- 9331188 TI - Frequency of optic disc drusen and size of the optic disc. PMID- 9331189 TI - How should we evaluate a cohort study? Study design. PMID- 9331191 TI - Excimer laser photorefractive keratectomy for high myopia: four-year experience with a multiple zone technique. AB - PURPOSE: The purpose of the study is to evaluate the results of the authors' 4 year experience with excimer laser photorefractive keratectomy (PRK) and multiple optical zone corneal ablation in highly myopic eyes. METHODS: The authors retrospectively evaluated 56 eyes of 44 patients (mean refraction, -11.3 diopters [D]; range, -5.75 to -24.5 D) who underwent PRK with a Visx Model 20/20 laser (Visx, Santa Clara, CA). Preoperative visual acuity of 20/40 or better was present in 46 eyes. Corneal ablation was divided into concentric optical zones (4, 5, and 6 mm), allowing corrections of up to 18 D, with a refractive goal of within -1 D from emmetropia in 49 eyes. A hand-held fixation system was always used, and a nitrogen-blowing system (NBS) was used in the first 21 eyes only. RESULTS: Before retreatment, the range of final cycloplegic refraction from emmetropia in eyes treated with NBS versus not was within +/-1 D in 6 (28.6%) and 15 eyes (44.1%), between -1.25 and -3 D in 5 (23.8%) and 14 eyes (41.1%), and more than -3 D in 10 (47.6%) and 5 eyes (14.7%), respectively. No lines of visual acuity were lost in 37 eyes (80.4%) with 20/40 or better visual acuity before surgery. Three eyes showed vision loss due to worsening of myopic maculopathy and one due to corneal haze. Correction stabilized within 9 months, and at a mean time of 25.6 months, the correction attained was of -8.5 +/- 3.6 D, achieving 90.3% of attempted correction. Eyes with preoperative myopia less than -10 D (n = 27) showed regression less than -1 D in 8 eyes (29.6%), between -1.25 and -3.00 D in 5 eyes (18.5%), and greater than -3.00 D in 1 eye (3.7%); eyes with more than 10 D (n = 29) regressed in 3 (10.3%), 6 (20.7%), and 1 eye (3.4%), respectively. Severe haze was observed in 11 eyes (19.6%) 3 months after surgery. Two eyes showed decentration greater than 1.5 mm. At last examination, night driving problems were reported by 12 (41.4%) of 29 patients evaluated who drive. CONCLUSIONS: After the NBS was eliminated, the multiple-zone technique achieved a long-term, stable 83.1% reduction of preoperative myopia. Patients with severe myopia appreciated reduction of most of the refractive defect, although perception of halos was noted by 16 patients. PMID- 9331190 TI - Results of phase III excimer laser photorefractive keratectomy for myopia. The Summit PRK Study Group. AB - OBJECTIVE: The purpose of the study is to determine safety and efficacy outcomes of excimer laser photorefractive keratectomy (PRK) for the treatment of mild-to moderate myopia. DESIGN: A prospective, multicenter, phase III clinical trial. PARTICIPANTS: A total of 701 eyes of 701 patients were entered in the study; 612 eyes were examined at 2 years after surgery. INTERVENTION: Intervention was photorefractive keratectomy using the Summit ExciMed UV200LA excimer laser (Summit Technology, Inc., Waltham, MA). The treatment zone diameter used was 4.5 mm in 251 eyes (35.8%) and 5 mm in 450 eyes (64.2%). Attempted corrections ranged from 1.50 to 6.00 diopters (D). MAIN OUTCOME MEASURES: Predictability and stability of refraction, uncorrected and spectacle-corrected visual acuity, refractive and keratometric astigmatism, corneal haze, contrast sensitivity, subjective reported problems of glare and halo, and patient satisfaction were the parameters measured. RESULTS: At 2 years, 407 (66.5%) eyes achieved 20/20 or better uncorrected visual acuity and 564 (92.5%) eyes achieved 20/40 or better visual acuity. Three hundred thirty-six (54.9%) eyes were within 0.5 D and 476 (77.8%) eyes were within 1.0 D of attempted correction. Stability of refraction improved with time; 86.8% of eyes were stable within 1.0 D from 6 to 12 months, 94% were stable from 12 to 18 months, and 96.3% were stable from 18 to 24 months. There was no evidence of progressive or late myopic or hyperopic refractive shifts. One hundred fourteen (18.6%) eyes gained 2 or more lines of spectacle corrected visual acuity, whereas 42 (6.9%) eyes lost 2 or more lines; however, of the latter, 32 (76.2%) had spectacle-corrected visual acuity of 20/25 or better and 39 (92.9%) eyes had 20/40 or better. Four hundred forty-two (72.2%) corneas were clear, 138 (22.5%) showed trace haze, 20 (3.3%) mild haze, 9 (1.5%) moderate haze, and 3 (0.5%) marked haze. On patient questionnaires, 87 (29.7%) patients reported worsening of glare from preoperative baseline; 133 (50.1%) reported worsening of halo symptoms from baseline. CONCLUSIONS: Photorefractive keratectomy appears effective for myopic corrections of -1.50 to -6.00 D. Uncorrected visual acuity is maximized in most eyes by 3 months, although some patients require between 6 months and 1 year to attain their best postoperative uncorrected visual acuity and some may require from 1 to 2 years for stabilization of refraction. Refraction stabilizes progressively without evidence of late myopic or hyperopic refractive shifts. Optical sequelae of glare and halo occur in some patients treated with a 4.5- or 5-mm treatment zone. PMID- 9331192 TI - A prospective evaluation of alcohol-assisted versus mechanical epithelial removal before photorefractive keratectomy. AB - OBJECTIVE: The purpose of the study is to compare alcohol-assisted versus mechanical debridement of the corneal epithelium before photorefractive keratectomy (PRK) for low-to-moderate myopia. DESIGN: A prospective study was performed on a group of consecutive patients operated on at the Massachusetts Eye and Ear Infirmary from February to April 1996 and followed for 6 months. PARTICIPANTS: Eighty patients (eyes) were divided in 2 groups: 40 alcohol and 40 mechanical. INTERVENTION: The patients underwent PRK for myopia (-1.5 to -7.5 diopters) with a Summit Apex excimer laser. The corneal epithelium was removed either with 20% ethanol or with a scalpel blade. MAIN OUTCOME MEASURES: The two groups were compared for epithelial removal time, epithelial defect size at the end of surgery, and rate of re-epithelialization. Uncorrected visual acuity (UCVA), refractive outcome, best-corrected visual acuity (BCVA), and subjective haze were measured at 4 days and at 1, 3, and 6 months. In an additional short term study, 40 patients (20 alcohol, 20 mechanical) had intraoperative pachymetry performed. RESULTS: Alcohol-assisted de-epithelialization was faster than mechanical debridement (107 [+/-20.6 standard deviation] versus 141 [+/-30.5] seconds [P < 0.0001]) and led to a more circumscribed and reproducible epithelial defect at the end of surgery (87,739 [+/-11,852] versus 103,518 [+/-33,942] square pixels [t test, P = 0.04; f test, P = 0.001]). At 4 days, 95% of the alcohol-treated patients had healed compared with 78% of the mechanically scraped patients (Fisher's exact test, P = 0.04). The alcohol group had a better UCVA at 4 days (logarithm of the minimum angle of resolution UCVA 0.36 [+/-0.22] versus 0.51 [+/-0.26]) and at 1 month (0.14 [+/-0.17] versus 0.22 [+/-0.16] [Mann Whitney U test, P = 0.02 and P = 0.03]) but equalized at 3 months (0.10 [+/-0.14] versus 0.13 [+/-0.16]) and at 6 months (0.11 [+/-0.15] versus 0.14 [+/-0.13] [Mann-Whitney U test, P = 0.23 and P = 0.34]). There was a trend toward less subjective haze in the alcohol-treated patients over the course of the study (area under the curve, 71.9 [+/-35.3] versus 87.9 [+/-33.8] [Mann-Whitney U test, P = 0.07]). The difference from target was equivalent in both groups at 6 months (-0.22 [+/-0.58] diopter in the alcohol group and -0.43 [+/-0.52] diopter in the mechanical group [t test, P = 0.14; f test, P = 0.57]). There were no differences in intraoperative pachymetry, corneal uniformity index as calculated from the corneal topography, and loss of BCVA between the two groups. CONCLUSIONS: Twenty percent ethanol is a simple, safe, and effective alternative to mechanical scraping before PRK and appears to be associated with a quicker visual rehabilitation. PMID- 9331193 TI - The effects of donor age on the outcome of penetrating keratoplasty in adults. AB - OBJECTIVE: The purpose of the study is to determine whether there is a higher incidence of complications in adult patients receiving corneas from pediatric donors compared to those receiving corneas from adult donors. DESIGN: The design is a follow-up of two matched cohorts. PARTICIPANTS: The outcome of penetrating keratoplasty in 29 adult patients (age 20 years of age and older) receiving pediatric donor corneas (range, 0-5 years) was compared to that of 29 control patients matched for recipient age and diagnosis who received adult donor corneas (range, 40-70 years). INTERVENTION: Chart review was performed. MAIN OUTCOME MEASURES: Graft rejection, postoperative keratometry, postoperative refractive cylinder, postoperative intraocular pressure, and graft failure due to rejection were measured. RESULTS: One or more allograft reactions occurred in 11 (37.9%) of 29 patients who received pediatric donor corneas compared to 2 (6.9%) of 29 patients who received adult donor corneas (P = 0.005, chi-square). There were a total of 20 rejection episodes in patients receiving pediatric donor corneas compared to a total of 5 rejection episodes in patients receiving adult donor corneas. The average postoperative keratometry was 46.1 diopters for the pediatric donor group and 44.0 diopters for the adult donor group (P = 0.03). There was no statistically significant difference in average refractive cylinder (P = 1.0), intraocular pressure (P = 0.26), or the incidence of graft failure due to rejection (P = 1.0) between the two groups. The average follow-up time for clear grafts was 58.3 months in the pediatric donor group and 59.9 months in the adult donor group. CONCLUSIONS: The incidence of allograft reactions and the postoperative corneal curvature is greater in adult eyes undergoing penetrating keratoplasty with young donor corneas compared to those undergoing penetrating keratoplasty with older donor corneas. There was no difference in the incidence of graft failure due to rejection between the two groups. PMID- 9331194 TI - Corneal transplantation in children with Peters anomaly and mesenchymal dysgenesis. Multicenter Pediatric Keratoplasty Study. AB - OBJECTIVE: The purpose of the study is to describe graft and visual outcomes of penetrating keratoplasty among young children with Peters anomaly and associated mesenchymal dysgeneses. DESIGN: The design was a multicenter retrospective analysis of the indications and outcome in pediatric keratoplasty. PARTICIPANTS: The records of all children aged 12 years and younger who underwent penetrating keratoplasty for mesenchymal dysgenesis between January 1975 and May 1993 at the participating centers were reviewed. MEASURES: The data were analyzed regarding graft survival and postoperative visual acuity. RESULTS: Forty-seven corneal transplants in 36 eyes of 29 patients with mesenchymal dysgenesis were studied. The majority of eyes operated on (30) had Peters anomaly (83%). Patients' mean age at the time of keratoplasty was 7 months. After a mean follow-up period of 38 months, 61% of eyes retained full graft clarity. One and 3-year survival rates were 79% (95% confidence interval [CI] = 65%-93%) and 62% (95% CI = 45%-79%), respectively. Postoperative corneal ulcers/nonhealing epithelial defects (P = 0.03), and additional noncorneal surgical procedures at the time of transplantation (P = 0.05) were associated with graft failure. Provision of postoperative optical aids (P = 0.01) was associated with better postoperative visual acuity levels. CONCLUSIONS: Penetrating keratoplasty for Peters anomaly and related mesenchymal dysgeneses in young children has a reasonable chance of success during the critical years of visual maturation and is associated with satisfactory visual results in one third to half the cases. The data suggest that complicated cases requiring additional surgical procedures have a worse prognosis. PMID- 9331195 TI - Outcome of acanthamoeba keratitis treated with polyhexamethyl biguanide and propamidine. AB - OBJECTIVE: This study investigates the clinical outcome of Acanthamoeba keratitis treated with polyhexamethyl biguanide (PHMB) and propamidine isethionate (Brolene). DESIGN: A retrospective review of all patients treated for Acanthamoeba keratitis between September 1992 and February 1995 was carried out. All patients were treated with PHMB 0.02% and propamidine 0.1% hourly for 3 days, the frequency reduced to four to six times daily according to clinical response. MAIN OUTCOME MEASURES: Age, gender, result of laboratory investigation, duration of disease before diagnosis, visual acuity (VA) pretreatment and post-treatment, need for keratoplasty, and presence of adverse reaction were measured. RESULTS: One hundred eleven cases were identified in 105 patients (60 male, 45 female; mean age, 32). Ninety-two percent of infections were in contact lens wearers. The clinical diagnosis was confirmed by corneal culture or histopathology in 64 cases (57.7%). The diagnosis was made "early" (within 28 days) in 65 cases (58.6%). Twenty-one (18.9%) were "intermediate" (28 days-2 months) and 20 (18%) were "late" (> 2 months) diagnoses. Overall post-treatment VA was 6/12 or better in the majority (88/111, 79.3%) of cases, and 18 (16.2%) had VA of 6/36 or worse. The VA of > or = 6/12 was achieved by 90.8% of the early, 71.4% of the intermediate, and 65% of the late groups. Clinical relapses occurred in 19 patients on reducing the therapy. Treatment toxicity was never serious and consisted only of stinging or superficial punctate keratopathy. Keratoplasty was indicated in only ten patients, and disease activity was controlled adequately in all patients before grafting. CONCLUSIONS: Combined treatment with PHMB and propamidine is well tolerated, nontoxic, and effective. Typically, visual outcome is favorable and the requirement for keratoplasty reduced markedly. PMID- 9331196 TI - Implications of early systemic therapy on the incidence of endogenous fungal endophthalmitis. AB - OBJECTIVE: In the past, evidence of endogenous fungal endophthalmitis has been used as a guide to initiating potentially toxic antifungal therapy in patients with systemic fungal infections. Recently, however, a trend has developed to provide patients with antifungal therapy at the first evidence of fungal infection. The authors' study evaluates the incidence of endogenous fungal endophthalmitis in this setting. DESIGN: The design is a retrospective review of the medical records of patients examined by the inpatient ophthalmology consultation service to rule out endogenous fungal endophthalmitis between January 1994 and April 1996 at the University of Michigan Hospitals, Ann Arbor, Michigan. PARTICIPANTS: Two hundred fourteen eyes of 107 patients with a diagnosis of systemic fungal infection were studied. INTERVENTION: A review of medical records was performed. MAIN OUTCOME MEASURES: The findings of the ocular examination, the presence of risk factors for disseminated fungal infection, the type of antifungal therapy, and the source and identity of the isolated fungus were recorded. RESULTS: The majority of patients examined had either fungal growth from blood cultures or evidence of deep tissue fungal infection. All patients in the study were at risk for fungal disease with each having at least one risk factor for disseminated fungal infection. Of the patients examined, 93.4% already were receiving systemic antifungal therapy at the time of ophthalmologic consultation. Only 3 (2.8%) of the 107 patients examined had chorioretinal findings consistent with early endogenous fungal endophthalmitis. None had intravitreous involvement, and the ocular findings did not change the course of therapy. CONCLUSIONS: Early systemic treatment of deep tissue fungal infection appears to dramatically decrease the incidence of endogenous fungal endophthalmitis. PMID- 9331197 TI - Orbital cysticercosis. AB - BACKGROUND: Human cysticercosis is secondary to an infestation by cysticercus cellulosae, the larval form of Taenia solium. Cysticercosis is endemic to regions with poor sanitation. The purpose of this report is to present a large series of patients with orbital cysticercosis and to discuss the current treatment. METHODS: A retrospective chart analysis of all patients with orbital cysticercosis from an urban practice in southern India was performed. The clinical features, the results of investigations, the therapies instituted, and the outcomes realized were recorded. RESULTS: Twenty patients diagnosed with orbital cysticercosis were identified (11 female and 9 male). Their ages ranged from 5 to 25 years with a mean age of 12.5 years. Nine patients manifested subconjunctival cysts. Eight were excised and 5 of these were densely adherent to the adjacent extraocular muscle (EOM). The remaining 11 patients had a cyst in a single EOM. The EOM cysts had proptosis, restricted motility, recurrent inflammation, and blepharoptosis. Two of the EOM cysts were excised surgically and four extruded spontaneously. Six patients with EOM cysts were treated medically: they all received oral corticosteroids and, additionally, five were given oral albendazole and one was given oral praziquantel. CONCLUSIONS: Excisional biopsy is recommended for subconjunctival cysticercosis. Idiopathic cystic myositis can present like EOM cysticercosis, but is differentiated by resolution with corticosteroid treatment. Medical therapy in orbital cysticercosis with oral albendazole and corticosteroids can arrest recurrent inflammation and improve ocular motility. PMID- 9331198 TI - Syphilis exposure in patients with uveitis. AB - PURPOSE: The purpose of the study is to determine the frequency of syphilis exposure in patients with uveitis, identify patient characteristics associated with serologic fluorescent treponemal antibody assays (FTA-ABS) reactivity, and examine the clinical implications of syphilis exposure in patients with uveitis. METHODS: A retrospective review of the records of 552 consecutive patients examined in the referral uveitis clinic of an urban eye hospital between January 1989 and January 1994 was performed. RESULTS: Forty-four (8%) of 552 consecutive patients with uveitis had serologic evidence of syphilis exposure on the basis of a strongly reactive serum FTA-ABS. Syphilis was presumed to be the sole cause of uveitis in 24 patients (4.3%) over the 5-year period. The racial demographic profile of those patients with serologic evidence of syphilis was consistent with the reported distribution of syphilis cases, but there were relatively few identifiable risk factors for sexually transmitted disease (including only three patients who were positive with human immunodeficiency virus). The choice of antibiotic treatment of these patients was variable and sometimes suboptimal. CONCLUSIONS: This study implicates syphilis exposure as a more common etiology of uveitis than did previous reports, advocates routine serum FTA-ABS testing of patients with uveitis, and indicates a need for a more aggressive role of the ophthalmologist in antibiotic treatment of patients with uveitis and syphilis exposure. PMID- 9331199 TI - Orbital compression syndrome in sickle cell disease. AB - BACKGROUND: Orbital complications are an uncommonly reported finding in sickle cell disease. METHODS: The authors review the reported orbital manifestations of sickle cell disease and discuss a patient with hemoglobin sickle beta(0) thalassemia in whom rapidly progressive bilateral orbital compression developed. RESULTS: Computed tomography of the orbits in a patient with fever, headache, orbital swelling, and optic nerve dysfunction displayed bilateral superior subperiosteal cystic masses. Surgical exploration showed bilateral liquefied hematomas, which were evacuated. Recovery was complete 13 days after surgery. A mild recurrence 14 months later resolved with conservative treatment. The literature contains 11 reports of 16 young patients with sickle cell disease (15 sickle cell disease [Hb SS] and 1 hemoglobin sickle cell disease [Hb SC]) with rapidly developing findings ranging from frontal headache, fever, and eyelid edema to bilateral complete orbital compression syndrome. Including our patient, 60% had orbital hemorrhage on computed tomography. Ten of 12 patients tested were found to have orbital bone marrow infarctions. Sixteen of 17 patients had complete recovery; 13 were treated conservatively and 4 surgically. Only 2 of 17 had recurrence. CONCLUSIONS: Orbital complications in sickle cell disease are unusual manifestations in which a vaso-occlusive process in the marrow space around the orbit results in frontal headache, fever, eyelid edema, and often orbital compression syndrome. Subperiosteal hematomas are common and appear to result from bone marrow infarctions. Appropriate management requires a thorough evaluation to exclude other hemorrhagic, infectious or neoplastic processes, as well as vigilant ophthalmic monitoring. Supportive care is effective, unless optic nerve dysfunction or large hematomas are present, which would indicate that surgical evacuation is warranted to prevent loss of vision and to speed recovery. PMID- 9331200 TI - Complications of motility peg placement for the hydroxyapatite orbital implant. AB - PURPOSE: The hydroxyapatite implant is an ocular motility implant designed to provide natural movement of the artificial eye. The movement of the prosthesis is maximized when the implant is coupled to the prosthesis via a peg. The purpose of this study is to determine the complications of the hydroxyapatite motility peg and the factors related to those complications. DESIGN: Retrospective review of 47 cases over 5 years. METHODS: A retrospective review was performed on all cases of hydroxyapatite motility peg placement. The technique of hydroxyapatite and peg placement, follow-up details, and complications of the peg were recorded. RESULTS: The complications of peg placement included peg extrusion in 26% (12 of 47), nonspecific conjunctivitis in 6% (3 of 47), audible click in 6% (3 of 47), temporary excessive conjunctival edema in 4% (2 of 47), and temporary excessive postoperative pain in 4% (2 of 47). There were no cases of infection, persistent pain, persistent edema, or discharge at peg site. The median time interval from peg placement to extrusion was 16 months (range, 1-52 months). The only statistically significant factor related to peg extrusion was age over 50 years (P = 0.04). There was a trend toward peg extrusion with use of a nonsleeved peg (versus sleeved peg) (P = 0.10). The extrusion rate was 32% (12 of 38) for nonsleeved pegs and 0% (0 of 9) for sleeved pegs. Factors unrelated to peg extrusion were patient sex, prior ocular surgery or radiotherapy, presence of giant papillary conjunctivitis, time interval from enucleation to peg placement, and degree of implant vascularization on magnetic resonance imaging. Of the 12 nonsleeved pegs that extruded, a sleeved peg system was subsequently successfully placed in 5 patients, a nonsleeved peg in 1 patient, and 6 patients remained without a peg system. CONCLUSIONS: Hydroxyapatite motility pegs have relatively few complications except for extrusion. The rate of extrusion can be minimized by employing a sleeved peg rather than a nonsleeved peg system. PMID- 9331201 TI - Adenoid cystic carcinomas of the lacrimal gland in childhood and adolescence. AB - OBJECTIVE: To see if there is a correlation between histologic features of these tumors and final outcome. DESIGN: A small series of cases of adenoid cystic carcinomas of the lacrimal gland in patients 18 years of age or younger were evaluated. PARTICIPANTS: A total of 11 cases of adenoid cystic carcinoma of the lacrimal gland in patients 18 years of age or younger found in the registry of Ophthalmic Pathology at the Armed Forces Institute of Pathology were studied. INTERVENTION: Histologic material obtained by excision of lacrimal gland tumors was evaluated for different morphologic parameters. Clinical follow-up information was reviewed. MAIN OUTCOME MEASURES: All cases were evaluated for proportion of a basaloid histologic pattern: necrosis, hemorrhage, mitotic count, and perineural, vascular, intraosseous, leptomeningeal, and optic nerve invasion. These parameters were examined for an association with the clinical follow-up that was obtained for eight of the patients (mean follow-up, 10 years; range, 2 14 years). RESULTS: Most of the patients were female (M:F = 2:9). Mean age was 14 years (range, 6.5-18 years). Of the patients with follow-up, 5 (62.5%) of 8 survived. Estimated survival rate at 15 years was 58% (Kaplan-Meier analysis). Survivors had 25% or less basaloid histology. Necrosis, hemorrhage, perineural invasion, and mitotic count were less prominent in survivors than in those who died of disease. Vascular invasion was seen only in fatal cases. CONCLUSIONS: Young patients with adenoid cystic carcinomas have a better prognosis than do adult patients, which may be due to their tumors having less aggressive histologic features. PMID- 9331202 TI - Porocarcinoma of the eyelid. AB - PURPOSE: The purpose of this report is to describe the clinical and histopathologic findings in a patient with porocarcinoma of the eyelid. METHODS: The case of a 68-year-old woman with a nodular lesion of the eyelid was studied, and the pertinent literature reviewed. RESULTS: No previous description of porocarcinoma of the eyelid was found in the literature. The present case presented with a nodular lesion of the right lower eyelid of 1-year duration. The tumor was completely excised. Microscopic study revealed ductal structures within lobules of tumor cells. CONCLUSIONS: Porocarcinoma is a rare cutaneous adnexal tumor arising from the eccrine secretory apparatus. Because the clinical behavior of this cancer includes deep invasion, regional lymphatic spread, and distant metastases, complete surgical excision is recommended, and should be verified by either conventional frozen sections or Mohs' micrographic sections. PMID- 9331203 TI - Subarachnoid fluid of the optic nerve in normal adults. AB - OBJECTIVE: The purpose of the study is to determine the amount of subarachnoid fluid of the optic nerve in normal adults using magnetic resonance (MR) imaging and to investigate whether the subarachnoid fluid is displaced in abduction as assumed by the 30 degree echographic test. DESIGN: The design was a prospective observation study. PARTICIPANTS: Twenty-one healthy headache-free adults participated. Ten (48%) were men and 11 (52%) were women with a mean age of 34.3 +/- 7.9 years. INTERVENTION: Both optic nerves were examined in primary and 45 degrees right and left gazes with T2 fast-spin echo fat-suppressed coronal MR imaging in quadrature head coil. Four MR images, 4 mm apart, starting from 4 mm posterior to the globe were obtained for both nerves simultaneously with an imaging time of 2 minutes and 24 seconds per gaze. MAIN OUTCOME MEASURES: Optic nerve and sheath diameters were measured. RESULTS: Mean nerve diameters were 3.2 +/- 0.4 mm anteriorly to 2.6 +/- 0.4 mm posteriorly, and mean sheath diameters were 5.2 +/- 0.9 mm anteriorly to 3.9 +/- 0.4 mm posteriorly. Optic sheath diameters did not change significantly in abduction or adduction. CONCLUSIONS: Magnetic resonance imaging can be used effectively to determine the amount of subarachnoid fluid of the optic nerve. In normal adults, the amount of optic nerve subarachnoid fluid is variable and may be substantial. The authors' MR findings show that optic nerve subarachnoid fluid is not displaced significantly with abduction or adduction. PMID- 9331204 TI - Radiation optic neuropathy after stereotactic radiosurgery. AB - PURPOSE: The purpose of the study is to report the occurrence of optic neuropathy after stereotactic radiosurgery for perichiasmal tumors. METHODS: Records of four patients with visual deterioration after stereotactic radiosurgery were reviewed, including clinical findings, neuroimaging results, and treatment methods. RESULTS: Optic neuropathy developed 7 to 30 months after gamma knife radiosurgery. All patients experienced an abrupt change in visual function. Clinical findings indicated anterior visual pathway involvement. Patterns of field loss included nerve fiber bundle and homonymous hemianopic defects. Gadolinium-enhanced magnetic resonance imaging (MRI) showed swelling and enhancement of the affected portion of the visual apparatus in three patients. Systemic corticosteroids were administered in all patients and one partially recovered. One patient also received hyperbaric oxygen without improvement. CONCLUSIONS: Although rare, optic neuropathy may follow radiosurgery to lesions near the visual pathways. Careful dose planning guided by MRI with restriction of the maximal dose to the visual pathways to less than 8 Gy will likely reduce the incidence of this complication. PMID- 9331205 TI - Eye injuries caused by bungee cords. AB - OBJECTIVE: Bungee cords are common workplace and household items that have many uses. Despite their utility, they represent a potential source of severe ocular injury. DESIGN: A retrospective review was conducted to identify patients presenting to an ocular emergency department with bungee cord-related eye injuries over a 42-month period. PARTICIPANTS: The authors identified 17 patients with bungee cord-related ocular injuries. INTERVENTION: In identified patients, the hospital inpatient and outpatient charts were reviewed in detail. MAIN OUTCOME MEASURES: Patient demographics, mechanism of injury, use of ocular protection, presenting and final best-corrected visual acuity, associated ocular injuries, therapeutic interventions, and anatomic results were noted. RESULTS: Fourteen (82%) of the patients had closed globe injuries and 3 (18%) had open globe injuries. A wide array of periocular, anterior segment, and posterior segment injuries were identified with hyphema being the most common associated ocular injury. Ten patients (59%) had a final visual acuity of 20/25 or better. Five patients (29%) had a final visual acuity that was less than or equal to 20/60, with three of these patients having a final visual acuity less than or equal to 4/200. Two of the three patients with open globe injuries had a final visual acuity of 20/60 or better, whereas one had no light perception. Poor visual outcome was associated with posterior segment involvement. CONCLUSIONS: The authors advocate the use of printed warnings on the packaging of bungee cords and extreme caution by those who use them. A modification in design and the use of certified safety glasses may help to decrease the incidence of bungee cord related ocular trauma. PMID- 9331206 TI - Long-term visual outcomes in patients with successful macular hole surgery. AB - OBJECTIVE: The purpose of the study is to determine the long-term visual outcomes in patients undergoing successful macular hole surgery. DESIGN: A consecutive series of eyes with an anatomically successful macular hole surgical result and at least 1 year postoperative follow-up information was identified and studied. Preoperative and postoperative visual acuities were measured in accordance with the Early Treatment Diabetic Retinopathy Study protocol. MAIN OUTCOME MEASURES: Visual acuity, improvement of visual acuity, and rate of final visual greater than or equal to 20/40 were measured. RESULTS: The median visual acuity increased from 20/125 before surgery to 20/50 1 year after surgery (93 eyes) and to 20/30 at 36 months after surgery (68 eyes). The trend for improvement in visual acuity after 1 year after surgery was statistically significant. The postoperative visual acuity was greater than or equal to 20/40 in 15 (17%) eyes at 3 months and 53 (78%) at 36 months. Before surgery, 12 (13%) eyes were pseudophakic, and 77 (83%) were pseudophakic at 36 months. Median visual acuity in the fellow eye was 20/32 at baseline and 20/32 at 36 months. The visual acuity in the study eye was better than in the fellow eye in 36 (39%) patients at 36 months after surgery. CONCLUSIONS: Visual acuity in patients after anatomically successful macular hole surgery continues to improve even beyond 1 year after surgery. Although substantial improvement occurs soon after cataract extraction, further improvement in visual acuity continues for 2 years thereafter. PMID- 9331208 TI - Insulin-like growth factor-1 retinal microangiopathy in the pig eye. AB - OBJECTIVE: Human recombinant insulin-like growth factor-1 (hrIGF-1), a ubiquitous angiogenic growth factor, was injected into the vitreous cavity of pigs to investigate the clinical and histopathologic consequences of supraphysiologic levels of this angiogenic growth factor on the retinal vasculature. DESIGN: Young male pigs were injected with 600 microg hrIGF-1 into the vitreous cavity and were observed with serial examinations by ophthalmoscopy, fundus photography, and fluorescein angiography for varying periods up to 6 months. In a separate set of experiments, a dose-response relation was explored in animals injected with varying doses of IGF-1. MAIN OUTCOME MEASURES: Histopathologic analysis included light and transmission electron microscopy and modified elastase digestion. Quantitative morphometric measurements were made of capillary basement membrane thickness and endothelial cell and pericyte densities of the retinal capillaries. RESULTS: Early clinical features of IGF-1-injected eyes included marked arteriolar tortuosity, vitreitis, and retinal vessel and optic nerve head vascular fluorescein leakage. By 4 weeks, hyperfluorescent dots consistent with microaneurysms appeared and increased in number until 8 weeks postinjection. Clinical findings did not change appreciably after 8 weeks. Elastase digestion showed microaneurysms of the retinal capillaries and no ischemia or pericyte ghosts. Quantitative analysis of the digested specimens showed increased endothelial density by 1 month after injection (P < 0.05). Transmission electron microscopic cross-sections of capillaries showed significant basement membrane thickening by 3 months (P < 0.05). Lower doses of IGF-1 showed fewer clinical and histopathologic changes, and no significant changes were noted with a single 6 microg injection. Suspending hrIGF-1 in acidic buffer produced less intraocular inflammation than use of bovine serum albumin at neutral pH. CONCLUSIONS: A single intravitreous injection of a large dose of hrIGF-1 produces a retinal microangiopathy that has a prolonged time of onset and remains stable from 2 to 6 months after injection. Some aspects of this angiopathy resemble diabetic retinopathy, suggesting growth factor effects in the morphologic vascular changes of diabetes. PMID- 9331207 TI - Central serous chorioretinopathy associated with inhaled or intranasal corticosteroids. AB - OBJECTIVE: The purpose of the study is to investigate the relationship between inhaled or intranasal adrenergic agonists and corticosteroids and the development of central serous chorioretinopathy (CSC). DESIGN: The medical records of three patients with CSC who were found to use inhaled adrenergic agents or corticosteroids or both were identified prospectively. A survey of members of the Retina, Macula, and Vitreous societies and the National Registry of Drug-Induced Ocular Side Effects identified three additional cases. RESULTS: Six patients with CSC were found to be chronic users of corticosteroid (four patients) or both beta adrenergic agonist and corticosteroid (two patients) metered dose inhalers or nasal sprays. In three cases, there was a close temporal correlation between the use of a corticosteroid nasal spray and the development of CSC. CONCLUSIONS: These findings suggest that, in patients who are susceptible, the periocular or systemic absorption of inhaled corticosteroids may be sufficient to produce CSC in humans, supporting previous hypotheses regarding the pathogenesis of the disorder. Further studies are needed to confirm this association and to determine whether inhaled adrenergic agents also contribute to the development of this disorder. Patients in whom CSC develops while using corticosteroid inhalers or nasal sprays should be alerted to the possible relationship between CSC and these agents. PMID- 9331209 TI - Fluorescent dots in fluorescein angiography and fluorescein leukocyte angiography using a scanning laser ophthalmoscope in humans. AB - PURPOSE: The purpose of the study is to disclose the nature of fluorescent dots and segments traditionally observed with fluorescein angiography (FA) using a scanning laser ophthalmoscope (SLO 101; Rodenstock, Munchen, Germany). The authors developed a new method, called fluorescein leukocyte angiography (FLA), to display directly the movement of leukocytes in human retinal vessels. METHODS: Fluorescein angiography was performed on two normal volunteers using a scanning laser ophthalmoscope and fluorescent dots and segments were observed. Fluorescein leukocyte angiography, using an injection of fluorescent buffy coat layer from which the fluorescent plasma and nonfluorescent erythrocytes have been removed externally, was performed on seven normal volunteers. Injection fluid smears were examined through a fluorescent microscope. Peripheral blood smears taken during midphase of FA and FLA also were examined. In addition, 15 early-phase FAs of central serous chorioretinitis (CSC) were studied retrospectively. RESULTS: In the FAs of normal volunteers, fluorescent dots were detected only in perimacular capillaries at early phase. Eight of the 15 CSC FAs examined showed both fluorescent dots and segments. In the FLAs, fluorescent dots were detected in whole retinal vessels for more than 30 minutes. Fluorescent segments were observed in FA but not in FLA. Injected fluid smears from one FLA showed fluorescent leukocytes and small platelets. However, in peripheral blood smears of the FLA, leukocytes and platelets were more visible and exhibited higher contrast than those of an FA due to background plasma fluorescence. The mean velocity of 21 flowing leukocytes in perifoveal capillaries was 1.37 +/- 0.35 mm/second in 2 FAs and that of 89 flowing leukocytes was 1.41 +/- 0.29 mm/second in 7 FLAs. CONCLUSIONS: The authors' observations suggest that fluorescent dots in scanning laser ophthalmoscope imaging are fluorescein-stained leukocytes, whereas fluorescent segments are the hyperfluorescent plasma that is located between rouleaux formations of erythrocytes. The velocity of the fluorescent dots could be measured in the perimacular capillaries by either FA or FLA; however, only FLA can display the flow of fluorescent leukocytes in large vessels. PMID- 9331210 TI - Visual function abnormalities and prognosis in eyes with age-related geographic atrophy of the macula and good visual acuity. AB - PURPOSE: Geographic atrophy (GA) may cause significant compromise of visual function, even when there still is good visual acuity (VA), because of parafoveal scotomas and foveal function abnormalities antedating visible atrophy. This study evaluates the visual function abnormalities at baseline and the 2-year worsening of VA and reading rate for eyes with GA compared with a group of eyes with drusen only. METHODS: Seventy-four eyes with GA and VA greater than or equal to 20/50 from a prospective natural history study of GA were included, as were 13 eyes with only drusen. Baseline visual function testing and 2-year VA and maximum reading rate are reported. RESULTS: The worsening of VA in decreased luminance and foveal dark-adapted sensitivity showed severe abnormalities for the GA group. Contrast sensitivity was significantly reduced for the eyes with GA. Half the eyes with GA, but none of the drusen eyes, had maximum reading rates below 100 words per minute. A scanning laser ophthalmoscope (SLO) measure of the scotoma near fixation combined with a measure of residual foveal function accounted for 54% of the variability in maximum reading rate in the eyes with GA. Of 40 eyes with GA observed for 2 years, half lost greater than or equal to 3 lines of VA and one quarter lost greater than or equal to 6 lines. The nine eyes with drusen with follow-up had no significant change in VA. Low foveal dark-adapted sensitivity, SLO measures of the scotoma within 1 degree of fixation, and low maximum reading rate were statistically significant risk factors for doubling of the visual angle. Significant reduction in maximum reading rates at 2 years was present for the eyes with GA. CONCLUSIONS: The eyes with GA with good VA have profound decreases in visual function, particularly in dim lighting and in reading. Half the eyes with GA had doubling in visual angle at 2 years after the baseline examination, whereas the drusen eyes remained essentially unchanged. Impaired visual function at baseline was predictive of an adverse outcome for the eyes with GA. PMID- 9331211 TI - Tube-shunt surgery versus neodymium:YAG cyclophotocoagulation in the management of neovascular glaucoma. AB - OBJECTIVE: To determine the relative effectiveness of neodymium:YAG (Nd:YAG) cyclophotocoagulation (CPC) and tube-shunt surgery on intraocular pressure (IOP) control in eyes with neovascular glaucoma (NVG). DESIGN: Retrospective, case-by case matched, comparative group study. PARTICIPANTS: Twenty-four patients with NVG treated with noncontact Nd:YAG-CPC were matched with 24 patients who underwent tube-shunt surgery. Matching criteria included the underlying disorder causing angle neovascularization, the lens status, and patient's age. INTERVENTIONS: Tube-shunt surgery or Nd:YAG-CPC. MAIN OUTCOME MEASURE: Postoperative IOP (IOP > or = 6 and < or = 25 mmHg), visual acuity, and presence of any postoperative complications. RESULTS: Satisfactory IOP control (IOP < or = 25 mmHg and > or = 6 mmHg) was achieved in 9 eyes (37.5%) treated with Nd:YAG-CPC compared with 16 eyes (66.7%) receiving a tube-shunt procedure (P = 0.04) over a mean follow-up of 16.9 +/- 14.6 and 15.2 +/- 11.8 months, respectively. In the matched pairs in both groups that had nonequivalent outcomes, the proportions with persistently high IOP or hypotony were both greater in the CPC group than in the tube-shunt group. The cumulative proportion of failure in the CPC group was 20.8% at 6 months, 35.4% at 1 year, and 71.2% at 3 years postoperatively. In the tube-shunt group, the cumulative proportions of failure at 6 months and 1 year were close to those in the CPC group (12.5% and 29.2%, respectively), but lower 3 years after surgery (43.3%). Eleven eyes (45.8%) in the CPC group lost light perception versus four eyes (16.7%) in the tube-shunt group. Complication rate was higher in the tube-shunt group. CONCLUSIONS: This study suggests that, in the management of NVG, tube-shunt surgery more frequently controls IOP in a satisfactory range, with less hypotony and less visual loss, than noncontact Nd:YAG-CPC. PMID- 9331212 TI - In vitro flow testing of glaucoma drainage devices. AB - OBJECTIVE: The study was intended to determine methods that could evaluate in vitro the flow characteristics of glaucoma drainage devices. DESIGN: Two test methods were used: (1) a gravity-driven flow test and (2) a syringe-pump-driven flow test. Eighteen devices, both valved and nonvalved, from 4 manufacturers were evaluated for their hydrodynamic resistance and the pressure at which the flow becomes 0. OUTCOME: Results show a wide variation in device performance, indicating a strong need for enhanced quality control procedures in the device manufacturing process. CONCLUSION: A gravity-driven flow test provides a reasonably quick test of both resistance and closing pressure, which might be useful as a manufacturing line test. The syringe-driven flow test requires more time but provides additional insight into device performance, and, therefore, might be useful as a design validation test. PMID- 9331213 TI - Comparative study of silicone versus acrylic foldable lens implantation in primary glaucoma triple procedure. AB - OBJECTIVE: To compare silicone versus acrylic foldable intraocular lens (IOL) implantation in primary glaucoma triple procedure (PGTP). DESIGN: Prospective, randomized. PARTICIPANTS: A total of 79 eyes of 79 primary open-angle glaucoma (POAG) patients in need of combined surgery were randomized to a silicone IOL group (36 eyes) and acrylic IOL group (43 eyes). INTERVENTION: The study eyes underwent PGTP, which consisted of primary trabeculectomy, phacoemulsification, and posterior chamber IOL implantation. Adjunctive mitomycin C (MMC) (0.5 mg/ml for 1 minute) was used selectively only in patients with one or more risk factors for filtration failure of PGTP. MAIN OUTCOME MEASURES: Snellen visual acuity, intraocular pressure (IOP), slit-lamp biomicroscopy, and number of glaucoma medications were measured, performed, or determined preoperatively and at regular intervals postoperatively. RESULTS: There were no significant differences in the mean number of postoperative glaucoma medications at 1, 2, 3, 4-6, and 9-12 months and at last follow-up (P > 0.05); mean change in corrected visual acuity best attained (P = 0.315) or at last follow-up (P = 0.223) between the silicone and acrylic groups. Both groups had significant decreases in mean IOP and mean number of medications postoperatively at all times (P < 0.05). However, the postoperative IOP > 25 mmHg and IOP spike > 5 mmHg above preoperative IOP during the first month were significantly higher in the acrylic group (P = 0.026). The mean postoperative IOP at 1 month in the acrylic group was also significantly higher than the silicone group (14.1 +/- 5.0, 11.2 +/- 3.9, P = 0.005). Conversely, there were no significant differences in mean postoperative IOP at 2, 3, 4-6, and 9-12 months and at last follow-up between the silicone and acrylic groups (P > 0.05). Suture removal or release occurred significantly more frequently in the acrylic IOL group during the first month and the first 2 months (48.8% and 60.5%) than the silicone group (25.0% and 36.1%, P = 0.030 and 0.031, respectively). There were no significant differences in postoperative complications or surgical interventions between the two groups (P > 0.05). CONCLUSIONS: During the first year following the PGTP with selective use of MMC, there were no significant differences in the medical dependency or visual outcomes or complications between the silicone and acrylic groups. Both groups attained significant decreases in IOP postoperatively. However, the mean IOP was significantly higher in the acrylic than the silicone group at 1 month postoperatively, and postoperative IOP > 25 mmHg and IOP spike > 5 mmHg above preoperative IOP were significantly greater in the acrylic group. There were significantly more suture releases in the acrylic IOL group than the silicone IOL group in the first 2 months postoperatively. PMID- 9331214 TI - Is there an association between migraine headache and open-angle glaucoma? Findings from the Blue Mountains Eye Study. AB - OBJECTIVE: To determine whether an association exists between migraine headache history and open-angle glaucoma (OAG). DESIGN: Population-based, cross-sectional study. PARTICIPANTS: Subjects were 3654 people aged 49 or older; 82% of permanent residents from an area west of Sydney participated. INTERVENTION: All participants underwent an interview and a detailed eye examination, including automated perimetry and stereo optic disc photography. MAIN OUTCOME MEASURES: Open-angle glaucoma was diagnosed in subjects with matching typical glaucomatous visual field defects and pathologic optic disc cupping, independent of intraocular pressure level. The diagnosis of migraine history (typical or nontypical) was based on participant responses to specific questions, consistent with International Headache Society criteria. RESULTS: Open-angle glaucoma prevalence increased exponentially with age, with rates of 0.4%, 1.3%, 4.7%, and 11.4% among persons aged less than 60 years, between 60 and 69 years, between 70 and 79 years, and 80 years or older, respectively. The frequency of reporting a past history of typical migraine headache declined with increasing age, with rates of 23.1%, 16.2%, 12.8%, and 10.4% for corresponding age groups. For all age groups combined, there was no significant association between typical migraine headache and OAG (odds ratio [OR], 1.3; 95% confidence interval [CI], 0.8-2.2), after multivariate adjustment. However, after stratifying into 10-year age groups, increased odds for OAG were found for people giving a history of typical migraine headache and aged 70-79 years (OR, 2.5; 95% CI 1.2-5.2), after adjusting for variables found associated with glaucoma. This association was marginally stronger for high-pressure OAG cases (OR, 2.7; 95% CI, 1.1-5.6). CONCLUSIONS: These data suggest the possibility of an association between history of typical migraine headache and OAG, which could be modified by age. PMID- 9331215 TI - A comparative study of two dose regimens of latanoprost in patients with elevated intraocular pressure. AB - OBJECTIVE: The purpose of the study was to determine whether latanoprost (13,14 dihydro-17-phenyl-18,19,20-trinor PGF2a-isopropyl ester), a new prostaglandin analogue that has been found effective in reducing intraocular pressure (IOP) in humans, is equally effective at lower concentrations than those currently employed. DESIGN AND PARTICIPANTS: Fifty patients with glaucoma or ocular hypertension were treated in a randomized, crossover, double-masked fashion with 1 drop of latanoprost (50 microg/ml once daily and 15 microg/ml twice daily) in the affected eye(s) for 3 weeks on each concentration. Tonometry was obtained at 8:00, 13:00, and 17:00 hours at baseline (untreated) and after 3 weeks on each concentration. Placebo (a buffer solution of latanoprost eye drop) was administered for complete masking of the study. RESULTS: Mean baseline (untreated) diurnal IOP for the entire sample was 24.7 mmHg. Intraocular pressure was reduced by 6.1 mmHg with latanoprost 15 microg/ml twice daily, and by 7.5 mmHg with 50 microg/ml once daily. Results with both regimens were significant (P < 0.001 each, Student's t-test). However, the 50 microg/ml dose was significantly more effective than the 15 microg/ml dose, with a difference of 1.4 mmHg (P < 0.001, ANOVA). Both dose regimens were well tolerated, with little, predominantly mild, ocular discomfort. The higher dose did not cause more hyperemia at 3 weeks than the lower one, i.e., the lower dose yielded a slightly higher score (1.8 mm) on the visual analogue scale (P < 0.29, ANOVA). CONCLUSIONS: Latanoprost administered at a concentration of 50 microg/ml once daily effectively reduces IOP in patients with elevated IOP. Administration of a lower concentration (15 microg/ml) twice daily is less effective, but still significant. PMID- 9331216 TI - Are motor enzymes bidirectional? PMID- 9331217 TI - Vertebrates have conserved capping protein alpha isoforms with specific expression patterns. AB - Capping protein (CP), a ubiquitous actin binding protein composed of an alpha and a beta subunit, is important for actin assembly and cell motility. Lower organisms have one gene and one isoform of each subunit. Chickens have two very similar alpha-subunit isoforms. To determine if vertebrates in general contain multiple alpha isoforms and if those alpha isoforms have conserved sequences, we isolated and analyzed alpha subunit cDNA's in mice and humans. Both mice and humans also have two alpha isoforms. Phylogenetic analysis of the alpha isoform sequences reveals that vertebrates have two highly conserved subfamilies, alpha1 and alpha2. The alpha1 and alpha2 subfamilies are very similar to each other but can be defined and distinguished from each other by a small number of key amino acid residues. In addition, 3' untranslated cDNA sequences are conserved within the isoform subfamilies. To investigate the function of the alpha isoforms, we examined their expression in mouse cells and tissues. Endothelial cells contain only the alpha2 isoform, and erythrocytes contain almost exclusively the alpha1 isoform. Most tissues have both alpha1 and alpha2 isoforms but the ratio of alpha1:alpha2 varies widely. Together, these findings support the hypothesis that the CP alpha isoforms have conserved, unique and essential roles in vertebrates. PMID- 9331218 TI - Visualization of single neurofilaments by immunofluorescence microscopy of splayed axonal cytoskeletons. AB - Treatment of cultured neurons with non-ionic detergents under certain conditions causes the axonal microtubules to splay apart from each other, allowing individual microtubules to be visualized by immunofluorescence microscopy [Brown et al., 1993, J. Cell Sci. 104: 339-352]. I have investigated whether axonal neurofilaments separate from each other under similar conditions. Cultures of dissociated dorsal root ganglion (DRG) neurons from fetal rats were treated with non-ionic detergent and fixed with formaldehyde. Neurofilaments were visualized by immunofluorescence microscopy using a polyclonal antiserum specific for NF-L. Treatment of the neurons with Triton X-100 or saponin caused filamentous structures to splay apart from each other along the entire length of the axon. Quantitative analysis of fluorescence intensity along the filamentous structures indicated that many of them represent single neurofilaments and that single and bundled neurofilaments can be distinguished based on their fluorescence intensity. The extent of this splaying phenomenon was dependent on time and detergent concentration. Temporal analysis indicated that short portions of single neurofilaments initially loop out from the axonal bundle and then subsequently splay apart further along their length and adhere to the polylysine/laminin coated substrate. The maximum observed length for a single axonal neurofilament was 183 microm in neurons after only 1 day in culture, which indicates that neurofilaments can attain remarkable lengths in these young cultured neurons. The splayed axonal cytoskeleton preparation described here allows individual axonal neurofilaments to be visualized by immunofluorescence microscopy, which is not possible in conventional preparations due to the dense packing of these polymers in axons. PMID- 9331219 TI - Localization of microtubules containing posttranslationally modified tubulin in cochlear epithelial cells during development. AB - In the adult gerbil inner ear, hair cell microtubules contain predominantly tyrosinated tubulin while supporting cell microtubules contain almost exclusively other isoforms. This cell-type specific segregation of tubulin isoforms is unusual, and in this respect the sensory and supporting cells in this sensory organ differ from other cells observed both in vivo and in vitro. Thus, we hypothesized there must be a shift in the presence and location of tubulin isoforms during development, directly associated with the onset of specialized functions of the cells. We describe the appearance and/or disappearance of tubulin isoforms in sensory hair cells and five different supporting cells (inner and outer pillar cells, Deiters cells, cells of Kolliker's organ, and cells of the tympanic covering layer) during development of the gerbil organ of Corti from birth to 14 days after birth. Tyrosinated tubulin was initially present in all cells and remained predominant in cells that decrease in number (Kolliker's organ and tympanic covering layer) and exhibit active processes such as secretion and motility (sensory cells). Posttranslational modifications occurred in the supporting cells in a time-dependent manner as the number and length of microtubules increased and development proceeded, but the establishment of elongated cell shape and polarity occurred prior to the appearance of acetylation, detyrosination, and polyglutamylation of tubulin. In the pillar and Deiters cells, posttranslational modifications progressed from cell apex to base in the same direction as microtubule elongation. In the pillar cells, posttranslational modifications occurred first at the apical surfaces. In the pillar cells, the appearance of acetylated tubulin was rapidly followed by the appearance of detyrosinated tubulin. In Deiters cells, the appearance of acetylated tubulin preceded the appearance of detyrosinated tubulin by one or more days. At onset of cochlear function, detyrosinated tubulin and acetylated tubulin had achieved their adult-like pattern, but polyglutamylated tubulin had not. PMID- 9331220 TI - Beta-thymosins from marine invertebrates: primary structure and interaction with actin. AB - The beta-thymosins are distributed throughout the vertebrate phyla, and all known vertebrate beta-thymosins bind actin monomers. To determine whether beta-thymosin like peptides function as actin-binding proteins in invertebrates, we fractionated perchloric acid extracts of the gonads of both the sea urchin, Arbacia punctulata, and the scallop, Argopecten irradians, and screened the fractions for proteins which could be crosslinked to actin. In each case a peptide was isolated which crosslinks to actin from both rabbit skeletal muscle and scallop cross-striated adductor muscle; both peptides were sequenced and each was found to consist of 40 amino acid residues, compared with 41-43 residues for the vertebrate beta-thymosins. The sequences of the scallop and sea urchin beta thymosins are 80% identical to each other, 75% identical to residues 1-40 of thymosin beta4, and 72-80% identical to residues 1-40 of other vertebrate beta thymosins. The sea urchin peptide was found to inhibit actin polymerization and nucleotide exchange. The affinity of the sea urchin peptide for rabbit muscle actin is apparently lower than that of thymosin beta4, since about twice the concentration of sea urchin peptide is required to give inhibition of actin polymerization or nucleotide exchange equivalent to thymosin beta4. PMID- 9331221 TI - Genetic evidence for a role of centrin-associated proteins in the organization and dynamics of the infraciliary lattice in Paramecium. AB - Within the superfamily of "EF-hand Ca2+-modulated proteins," centrins constitute a family of cytoskeletal proteins that are highly conserved from lower eukaryotes to man. Their cytoskeletal specialization is manifest in their capacity to form filamentous contractile arrays of various shapes and functions and by their association with microtubule organizing centres (MTOCs). While the latter property has been conserved throughout the evolution of eukaryotes, centrin-based contractile structures are only found in protists where they form arrays of widely diverse organization and function. In the ciliate Paramecium tetraurelia, three centrin genes have been characterized, which may be part of a larger centrin gene family [Madeddu et al., 1996: Eur J. Biochem. 238:121-128]. The products of these genes were originally identified as components of the infraciliary lattice, a contractile cytoskeletal network [Garreau de Loubresse et al., 1991: Biol. Cell 71:217-225]. We show here that centrins are localized not only in this lattice but also in basal bodies and in the cord, a filamentous structure associated with the oral apparatus. We demonstrate that in the infraciliary lattice, but not in basal bodies, centrins are associated with high molecular-weight proteins (ca. 350 kD). Their role in the biogenesis of the infraciliary lattice is documented by cytological and biochemical properties of the mutant "demaille" (dem1) characterized by altered centrin-associated proteins and abnormal organization and dynamics of the infraciliary lattice. PMID- 9331222 TI - Co-ordinate regulation of the cytoskeleton in 3T3 cells overexpressing thymosin beta4. AB - In several cell types, short-term increases in the concentration of the G-actin sequestering peptide thymosin-beta4 (Tbeta4) cause the disassembly of F-actin bundles. To determine the extent of cell adaptability to these reductions in F actin, we overexpressed Tbeta4 in NIH 3T3 cells. In cell lines with Tbeta4 levels twice those of vector controls, G-actin increased approximately twofold as expected. However, F-actin did not decrease as in short-term experiments but rather also increased approximately twofold so that the G-F ratio remained constant. Surprisingly, the cytoskeletal proteins myosin IIA, alpha-actinin, and tropomyosin also increased nearly twofold. These increases were specific; DNA, total protein, lactic dehydrogenase, profilin, and actin depolymerizing factor levels were unchanged in the overexpressing cells. The Tbeta4 lines spread more fully and adhered to the dish more strongly than vector controls; this altered phenotype correlated with a twofold increase in talin and alpha5-integrin and a nearly threefold increase in vinculin. Focal adhesions, detected by indirect immunofluorescence with antivinculin, were increased in size over the controls. Northern blotting showed that mRNAs for both beta-actin and vinculin were increased twofold in the overexpressing lines. We conclude that 1) NIH 3T3 cells adapt to increased levels of G-actin sequestered by increased Tbeta4 by increasing their total actin so that the F-actin/G-actin ratio remains constant; 2) these cells coordinately increase several cytoskeletal and adhesion plaque proteins; and 3) at least for actin and vinculin, this regulation is at the transcriptional level. We therefore propose that the proteins of this multimember interacting complex making up the actin-based cytoskeleton, are coordinately regulated by factors that control the expression of several proteins. The mechanism may bear similarities to the control of synthesis of another multimember interacting complex, the myofibril of developing muscle cells. PMID- 9331223 TI - Kinase and phosphatase inhibitors cause rapid alterations in microtubule dynamic instability in living cells. AB - To examine whether microtubule dynamic instability can be rapidly regulated during interphase, we used video-enhanced differential interference contrast (DIC) microscopy to observe individual microtubules at the periphery of living newt lung epithelial cells. Microtubules were observed before and after perfusion with either the phosphatase inhibitor okadaic acid or the kinase inhibitors staurosporine or olomoucine. Addition of these inhibitors caused rapid changes in dynamic instability. Thirty to sixty seconds after perfusion with 0.2-1 microM okadaic acid, a 1.5-fold increase in elongation velocity and small increases in catastrophe and rescue frequencies were observed. In contrast, treatment with 40 200 nM staurosporine decreased microtubule elongation and shortening velocities approximately 2-fold, and catastrophes were slightly more frequent. Olomoucine, at 100 microM, had similar effects. Transition dynamics were further examined by probabilistic analysis, which showed that microtubules become more likely to undergo catastrophe as they elongated and more likely to undergo rescue as they shortened, an effect previously called microtubule "memory." This memory effect for catastrophes was observed in untreated and okadaic acid-treated cells but was abolished by staurosporine or olomoucine. In contrast, the memory effect for rescue was unaffected by these treatments, suggesting that catastrophe and rescue proceed via distinct, multistep mechanisms. Overall, these results demonstrate that microtubule assembly regulators can be altered rapidly by inhibition of either kinases or phosphatases and suggest that, in the absence of inhibitors, these regulators exist in a dynamic equilibrium between phosphorylated and dephosphorylated states. PMID- 9331224 TI - Are we throwing out the baby? PMID- 9331225 TI - Changes in facial movement after maxillary osteotomies. AB - PURPOSE: Determine changes in facial movement while smiling after maxillary Le Fort I osteotomies. MATERIALS AND METHODS: Twenty patients (ages 15 to 38) treatment-planned for maxillary Le Fort I osteotomies were divided into two groups. Group A consisted of 10 patients who underwent superior and/or posterior positioning of the maxilla. Group B consisted of 10 patients who underwent anterior and/ or inferior repositioning of the maxilla. All patients underwent preoperative and postoperative (3 to 8 months) videographic analysis of a maximal closed mouth smile by the Johnson Maximal Static Response Assay, evaluating four landmarks around the mouth and nose (alar base--A, cheilion--C, labrale superioris--Ls, and intermediate between cheilion and labrale superioris--Im). RESULTS: Group A was noted to have a statistically significant decrease in movement of the face at points C and Im. No significant change was seen for points Ls and A. Group B was noted to have a statistically significant increase in movement of the face at point A, C, and Im. Point Ls was also found to increase, however not significantly. CONCLUSION: Surgical repositioning of the maxilla anteriorly and/or inferiorly lengthens the facial musculature resulting in an increase in facial movement while smiling. Likewise surgically repositioning the maxilla superiorly and/or posteriorly reduces the length of the facial musculature, resulting in a decrease in facial movement while smiling. PMID- 9331226 TI - Freehand full-thickness grafting for facial defects: a review of methods. AB - PURPOSE: This article reviews the use of full-thickness skin grafts for closure of facial defects. PATIENTS AND METHODS: In almost 3 years, 30 patients had full thickness skin grafting after removal of premalignant or malignant facial skin lesions. The most common graft harvest sites included the preauricular and postauricular, neck, and supraclavicular areas. RESULTS: Few complications were seen except for rare surface necrosis and depression of the grafted site, and the esthetic results were generally satisfactory. CONCLUSION: Full-thickness skin grafts offer a reliable alternative to the use of flaps in selected cases. PMID- 9331227 TI - Mandibular anterior ridge extension: a modification of the Kazanjian vestibuloplasty technique. AB - PURPOSE: A modification of the secondary epithelization vestibuloplasty technique described by Kazanjian that eliminates the sharp V in the depth of the extended vestibule and counteracts shallowing of the sulcus is presented. PATIENTS AND METHODS: Ten consecutive patients indicated for anterior mandibular secondary epithelization vestibuloplasty were treated. A bipedicled mucosal flap was developed in the labioalveolar mucosa for lining the extended vestibular depth. A comparison was made of the vestibular depth measured from the crest of the ridge to the junction of the attached mucosa both preoperatively and postoperatively. RESULTS: Healing of raw surfaces was uneventful. The mean preoperative anterior mandibular vestibular depth was 3.5+/-1.1 mm. After 6 months, the mean anterior mandibular vestibular depth was 9.2+/-1.7 mm, a statistically significant difference (P < .05). The mean gain in vestibular depth was 5.7+/-2.2 mm. CONCLUSION: Overcorrection is unnecessary with this modification. Elimination of the sharp V in the extended vestibular depth enables denture fabrication with better flange extension and improved oral hygiene. PMID- 9331228 TI - Amnestic and anxiolytic effects of alprazolam in oral surgery patients. AB - PURPOSE: This study was designed to identify a dose of alprazolam that would reduce anxiety associated with oral surgery without causing accompanying memory impairment. PATIENTS AND METHODS: Thirty-six subjects in experiment 1 and 48 subjects in experiment 2 were pretested on a computerized memory battery to establish baseline performance. Subjects were then randomly assigned to receive placebo, 0.25 mg, or 0.75 mg oral alprazolam (experiment 1) or placebo, 0.25 mg, 0.50 mg, or 0.75 mg oral alprazolam (experiment 2). Forty-five minutes after the double-blind administration of alprazolam, subjects were given a second memory battery. The memory batteries tested story recall and recognition and word recall and recognition. Subjects in experiment 2 subsequently underwent oral surgery for the removal of one to four molars. The subjects completed anxiety questionnaires both before and after surgery. RESULTS: The 0.75-mg and 0.50-mg doses, but not the 0.25-mg dose, impaired word recall. The 0.75-mg dose also impaired story recall and recognition. The proportion of subjects reporting moderate to high anxiety during oral surgery decreased with increasing doses of alprazolam. Multiple regression indicated that the 0.75-mg alprazolam dose significantly decreased anxiety during oral surgery. The 0.25-mg and 0.50-mg doses also tended to reduce anxiety, but beta values for these doses did not reach significance. CONCLUSIONS: These findings indicate that alprazolam produces memory impairment at the dosages necessary to produce clinically significant anxiolysis during oral surgery. PMID- 9331229 TI - Computerized cephalometric evaluation of orthognathic surgical precision and stability in relation to maxillary superior repositioning combined with mandibular advancement or setback. AB - PURPOSE: A computerized, cephalometric, orthognathic surgical program (TIOPS) was applied in orthognathic surgical simulation, treatment planning, and postoperatively to assess precision and stability of bimaxillary orthognathic surgery. PATIENTS AND METHODS: Forty consecutive patients with dentofacial deformities requiring bimaxillary orthognathic surgery with maxillary superior repositioning combined with mandibular advancement or setback were included. All patients were managed with rigid internal fixation (RIF) of the maxilla and mandible and without maxillomandibular fixation (MMF). Preoperative cephalograms were analyzed and treatment plans produced by computerized surgical simulation. Planned, 5-week postoperative and 1-year postoperative maxillary and mandibular cephalometric-positions were compared. RESULTS: In the mandibular advancement group, the anterior maxilla was placed too far superiorly, with an inaccuracy of 0.4 mm. The posterior maxilla and the anterior mandible were placed in the planned positions. The lower posterior part of the mandibular ramus was placed too far anteriorly, with an inaccuracy of 2.0 mm. However, the mandibular condyles were accurately placed. In the setback group, the anterior maxilla was placed too far superiorly and posteriorly, with a vertical and sagittal inaccuracy of 1.0 mm and 0.7 mm, respectively. The posterior part of the maxilla was placed in a posterior position with an inaccuracy of 1.9 mm. The anterior mandible was placed too far anteriorly with an inaccuracy of 0.9 mm. The lower posterior part of the mandibular ramus was placed in a posterior position with an inaccuracy of 0.9 mm. However, the mandibular condyles were accurately placed. The statistical analysis of the 1-year stability data showed that the maxilla had moved 0.3 mm posteriorly in the advancement group and the lower incisors had moved 0.8 mm superiorly. No other significant positional maxillary or mandibular changes were found. In the setback group, the maxilla had moved 0.5 mm posteriorly, the anterior mandible 0.5 mm anteriorly, and the lower incisors 0.7 mm superiorly. No significant positional changes were seen in the mandibular ramus. CONCLUSION: The TIOPS computerized, cephalometric, orthognathic program is useful in orthognathic surgical simulation, planning, and prediction, and in postoperative evaluation of surgical precision and stability. The simulated treatment plan can be transferred to model surgery and finally to the orthognathic surgical procedures. The results show that this technique yields acceptable postoperative precision and stability. PMID- 9331230 TI - Surgical treatment of fractures of the edentulous mandible. AB - PURPOSE: This retrospective study presents treatment alternatives for fractures of the edentulous mandible. METHODS: A chart review of 34 patients with fractures of the edentulous mandible was performed. Patients were followed with clinical and radiographic examinations. RESULTS: Five different treatment groups were used, ranging from closed treatment to bone graft augmentation and immediate placement of dental implants. Twenty-five patients showed good bony union. Frequent complications were encountered, including pseudarthrosis, fractured plates, and persistent dysesthesia. Augmentation was more stable when implants were placed simultaneously. CONCLUSION: Treatment needs to be based on the type and location of the fracture and the degree of atrophy. Augmentation in combination with implants appears to be a good treatment for fractures of the edentulous mandible. PMID- 9331231 TI - Bone grafting the piriform aperture deformity in isolated cleft lip patients: indication, technique, and results. AB - PURPOSE: This study was undertaken to determine the effect of a bone graft in the piriform aperture on the nasal deformity and orthodontic treatment of the cleft side teeth in isolated cleft lip patients. PATIENTS AND METHODS: All primary cleft lip repair was done 3 months after birth. Nine patients, four female and five male, with a mean age of 12.5 years (range, 8.2 to 24.8 years) and with a repaired cleft lip, were bone grafted between 1992 and 1996. The mean postoperative period was 2 years (range, 1 to 4 years). An iliac crest bone graft was placed in the piriform aperture deformity on the side of the cleft lip. The improvement in the nasal symmetry and angulation of the cleft side teeth were evaluated. The eight growing cleft lip patients (mean age, 11 years; range, 9 to 13 years) were compared with a control group of eight healthy growing children (mean age, 11 years; range, 9 to 13 years). The improvement of nasal symmetry was measured by the formula of the lobule portion of the columella index preoperatively and postoperatively. RESULTS: The mean lobule portion of the columella index preoperatively was 41.8% (SD, 4.4%; SE of Mean, 1.5%) and postoperatively was 44.2% (SD, 4.9%; SE of Mean, 1.6%) (P > .006, t-test for paired samples). The angulation of the cleft side teeth was improved by orthodontic treatment. CONCLUSION: Bone grafting the piriform aperture deformity results in a stable result and improves nasal symmetry and the angulation of the cleft side teeth. PMID- 9331232 TI - Accuracy of fine needle aspiration biopsy in head and neck tumors. AB - PURPOSE: Fine-needle aspiration biopsy (FNAB) is frequently used in the diagnosis of lesions occurring in the head and neck region. This study evaluated the correlation between the findings on FNAB and the histological findings observed after surgery. MATERIALS AND METHODS: A review of 218 patients who underwent FNAB of a head or neck tumor was performed. Cytological reports were classified into the following diagnostic categories: negative or positive for malignant cells and unsatisfactory. False-positive, false-negative, true-positive (sensitivity), and true-negative (specificity) rates were calculated. RESULTS: Twelve specimens did not allow an adequate diagnosis (5.5%). Among benign tumors, 96.2% of the cases were correctly diagnosed, and 3.8% were nondiagnostic specimens. Among malignant tumors, 86.4% of cases were correctly identified. There were two (3.4%) false negatives and six (10.2%) nondiagnostic specimens, with a total false-negative rate of 13.6%. CONCLUSIONS: Sampling errors present a minor problem with FNAB. Most nondiagnostic or incorrect specimens were caused by nonhomogenous lesions, with poor placement of the needle and an insufficient amount of aspirated material. FNAB is a useful modality for the diagnosis of head and neck masses. PMID- 9331233 TI - Repair of traumatic orbital wall defects with nasal septal cartilage: report of five cases. AB - PURPOSE: This article reports the use of nasal septal cartilage for the repair of traumatic orbital wall defects. PATIENTS AND METHODS: Five patients with disruption of the orbital wall after facial trauma were included in this retrospective review. All of the patients underwent open reduction with internal fixation of the fractures as well as repair of the orbital wall defect with autogenous septal cartilage. RESULTS: Nasal septal cartilage was used in four cases of orbital floor defect and one case of orbital roof defect. All of the cases were successfully treated by restoration of the orbital wall continuity. CONCLUSION: Nasal septal cartilage is a readily accessible autogenous material that can be easily harvested with minimal donor site morbidity, and it should be considered when an autogenous orbital implant is needed for the repair of a traumatic orbital wall defect. PMID- 9331234 TI - The effect of hyperbaric oxygen on irradiated oral tissues: transmucosal oxygen tension measurements. AB - PURPOSE: This study measured the effect of hyperbaric oxygen (HBO) treatment on transmucosal oxygen tension in irradiated human oral mucosa. PATIENTS AND METHODS: Ten patients received 30 dives of HBO as part of their treatment for mandibular osteoradionecrosis. A noninvasive, nonheated oxygen electrode was used to measure the tissue surface transmucosal oxygen tension directly on the attached gingiva. Measurements were done before, during, and after HBO treatment. The normal level of gingival surface transmucosal oxygen tension was measured in five healthy volunteers. RESULTS: During HBO treatment, the transmucosal oxygen tension increased significantly after five dives of HBO (P < .05). After 30 dives, the increases were from a mean of 50% to a mean of 86% of the transmucosal oxygen tension of normal healthy gingiva. CONCLUSION: An increase in the transmucosal oxygen tension is based on neo-angiogenesis. Patients with subischemic tissues, such as the study population with postirradiation mucosal and osseous necrosis, therefore may benefit from treatment with HBO. PMID- 9331235 TI - Markers for macrophage and osteoclast lineages in giant cell lesions of the oral cavity. AB - PURPOSE: Giant cell lesions of the oral cavity are a well recognized entity. However, the histogenesis of these lesions is still the subject of controversy, with support for both histiocyte/macrophage and osteoclast origins being found in the literature. This study evaluated a set of peripheral giant cell lesions (PGCLs) and central giant cell lesions (CGCLs) for characteristics of both cell types to address this dilemma. MATERIALS AND METHODS: Detection of histiocyte/macrophage characteristics was accomplished immunohistochemically by evaluating for markers specific for this cell type, namely alpha-1 antichymotrypsin (1 -ACT) and factor XIIIa antibodies. Detection of osteoclast characteristics made use of the fact that osteoclasts possess a unique enzyme, tartrate-resistant acid phosphatase, which can be appreciated by histochemical procedures. RESULTS: A large percentage of the multinucleated cells stained with the 1-ACT (38.08% in PGCLs and 15.84% in CGCLs), while only isolated cells stained for factor XIIIa (1.20% PGCLs, 0.99% CGCLs). Isolated stromal cells also were stained. Virtually all multinucleated cells reacted with the tartrate resistant acid phosphatase stain (99.26% PGCLs, 98.34% CGCLs), as did a number of the mononuclear stromal cells. CONCLUSIONS: This study supports the contention that GCLs of the oral cavity may arise from precursor cells related to the granulocyte/macrophage line, and may originate from mononuclear cells that express markers for both macrophages and osteoclasts. PMID- 9331236 TI - The effect of hyaluronic acid on experimental temporomandibular joint osteoarthrosis in the sheep. AB - PURPOSE: The purpose of this study was to test the effect of repeated injections of hyaluronic acid (HA) on the sheep model of osteoarthrotic temporomandibular joint (TMJ) disease. MATERIALS AND METHODS: Bilateral osteoarthrosis (OA) was induced in the TMJs of six sheep. HA was injected into one joint on 7, 10, 14, 17, and 21 days postoperatively. Normal saline was injected into the contralateral joint. Three sheep were killed at 1 month and 3 at 3 months. The joints were removed and examined macroscopically and histologically. A special scoring system was applied following the modified Mankin's score to evaluate the histologic changes. RESULTS: The control group showed severe osteoarthrotic changes in the condyle, deviation in form from normal morphology, and marked marrow fibrosis. The HA-treated group showed less deviation from normal condylar morphology. The histologic scores at 1 month were HA 12.6, control 24.2 (P < .001), and at 3 months were HA 6.9, control 18.9 (P < .001). There was a significant difference in osteoarthrotic changes between HA-treated and control TMJs, with the HA-treated TMJs having less severe changes. CONCLUSION: Repeated intraarticular injections of HA into a sheep TMJ with experimentally induced OA minimizes the extent of osteoarthrotic change when compared with the control joint. Thus, HA may have a role in preventing the progression of TMJ OA. PMID- 9331237 TI - Treacher Collins syndrome: perspectives in evaluation and treatment. PMID- 9331239 TI - Aneurysmal bone cyst-"plus": a report of three cases. PMID- 9331238 TI - Large perimandibular swelling. PMID- 9331240 TI - Mandibular reconstruction using the anterior part of ascending ramus: report of two cases. PMID- 9331241 TI - Oral squamous cell carcinoma as a second primary malignancy in a patient with previously diagnosed osteosarcoma of the femur. PMID- 9331242 TI - Entomophthora coronata infection of the paranasal sinuses: report of a case. PMID- 9331244 TI - The fasciocutaneous cervicopectoral rotation flap for lower cheek reconstruction: report of three cases. PMID- 9331243 TI - Primary non-Hodgkin's lymphoma of the mandible: a case report. PMID- 9331245 TI - Modified procedure for cephalic vein transposition for free flap salvage: report of case. PMID- 9331246 TI - Camel bite injuries of the orofacial region: report of a case. PMID- 9331247 TI - Osteosarcoma of the maxilla: report of a case and review of the literature concerning metastasis. PMID- 9331248 TI - Sialadenoma papilliferum of the oral cavity: report of a case and literature review. PMID- 9331249 TI - Protective guide plate to aid in downfracture and positioning of the maxilla after Le Fort I osteotomy. PMID- 9331250 TI - Re-extraction HBO: empiricism or science? PMID- 9331251 TI - Ultrasound for the treatment of osteoradionecrosis. PMID- 9331252 TI - Another shot at gun control. PMID- 9331253 TI - Introduction of George B. Benedek 1997 recipient of the Proctor Medal. PMID- 9331254 TI - Cataract as a protein condensation disease: the Proctor Lecture. PMID- 9331255 TI - Transforming growth factor-beta receptor expression in human cornea. AB - PURPOSE: Limbal basal cells and corneal endothelial cells appear to be inhibited in the G1 phase of the cell cycle. As a preliminary to determining whether transforming growth factor-beta (TGF-beta) might mediate this inhibition, investigation was made to determine whether human corneal and limbal cells express TGF-beta receptor types I (RI), II (RII), and III (RIII). METHODS: Corneas from eight human donors, aged stillborn to 85 years, were fresh frozen, cryostat sectioned, and prepared for indirect immunofluorescence localization of RI, RII, and RIII, using an established protocol. Corneas from donors 50 years of age or older were used to prepare RNA from the epithelium and endothelium. Reverse transcription-polymerase chain reaction was conducted using primers specific for each TGF-beta receptor type. RESULTS: Immunolocalization patterns for RI, RII, and RIII were similar, regardless of donor age. Binding of RI and RII antibodies was barely detectable in central corneal epithelium; however, most limbal basal cells stained positively for RI and RII. All layers of central corneal epithelium and the suprabasal layers of the limbus stained positively for RIII, whereas staining for this receptor was markedly decreased in limbal basal cells. Corneal endothelium bound the antibody for all three TGF-beta receptor types. In the same tissue sections, antibody staining for the RIII protein was more intense in corneal endothelial cells than in limbal basal cells. Polymerase chain reaction product for RI, RII, and RIII was detected in the epithelium and in the endothelium. CONCLUSIONS: Limbal basal cells and corneal endothelial cells expressed mRNA and protein for TGF-beta receptor types I, II, and III, suggesting that both cell types can transmit a TGF-beta-induced signal. These two cell types may differ in their relative response to those TGF-beta isoforms that require binding to RIII for signal transduction, in that staining intensity for RIII was relatively low in limbal basal cells compared with that in the endothelium. That limbal basal and corneal endothelial cells express receptors for TGF-beta suggests that this cytokine could mediate G1 phase arrest in these two cell types. PMID- 9331256 TI - A new, simple, nonradioactive, nontoxic in vitro assay to monitor corneal endothelial cell viability. AB - PURPOSE: This study was designed to determine whether Alamar blue could be used to evaluate corneal endothelial cell viability in vitro. METHODS: Alamar blue incorporates a proprietary redox indicator that changes color in response to metabolic activity. Primary rabbit endothelial cells were subcultured on 96-well plates at densities ranging from 1,250 to 40,000 cells per well. After 12 hours' incubation, Alamar blue was added to each well and absorbance measured hourly from 1 to 9 hours. Sodium azide-killed cells were used as a control. Alamar blue conversion was also compared with [3H]-thymidine incorporation in the presence or the absence of mitomycin C. RESULTS: Alamar blue reduction demonstrated endothelial cell viability at all cell concentrations compared with that in killed-cell controls. The reduction varied proportionately with cell number and time, showing clearly significant differences. Conversely, [3H]-thymidine uptake demonstrated minimal DNA synthesis and little or no ability to distinguish cell number or viahility. CONCLUSIONS: Alamar blue reduction measures endothelial cell viability and can readily differentiate cell concentrations. It demonstrates several advantages over [3H]-thymidine: It can assay nonproliferating endothelial cell metabolism, it allows rapid assessment of large numbers of samples, it can differentiate endothelial cell concentrations, it is nontoxic, it is nonradioactive and allows for simple disposal, it is less costly, and it allows for continuous monitoring of endothelial cell metabolism and viability. PMID- 9331257 TI - Cryopreservation of rabbit corneas in dimethyl sulfoxide. AB - PURPOSE: To minimize the injury to endothelial cells during cryopreservation of rabbit corneas with dimethyl sulfoxide. METHODS: Rabbit corneas were cryopreserved using 20% wt/wt dimethyl sulfoxide (Me2SO), added and removed in stages to maintain the osmotically induced excursions in cell volume to within +/ 40% of their isotonic volume. The vehicle solution, cooling rate, and conditions of storage used were those already reported to be optimal for endothelial cell survival after exposure to low temperatures. Survival was assessed by confocal microscopy with vital staining and by the ability of the endothelium to control stromal hydration during 3 hours of normothermic perfusion. The effect of temperature of addition and removal of Me2SO (room temperature [RT] or 2 degrees C) on endothelial viability also was measured. RESULTS: After thawing, all the cryopreserved corneas appeared structurally intact when assessed by vital staining and could limit stromal swelling during subsequent normothermic perfusion. Analysis of the rate of stromal swelling during the first 1.5 hours of normothermic perfusion indicated a substantial benefit when the Me2SO was removed at RT. Adding and removing the Me2SO at RT, which allowed a briefer exposure to Me2SO before cooling, resulted in better structural integrity of the endothelial layer than when the addition of cryoprotectant took place on ice. CONCLUSIONS: These results demonstrate the importance of osmotic stresses in the generation of injury to corneal endothelium during cryopreservation and the possibility of eventual successful cryopreservation of this tissue. PMID- 9331258 TI - Reduction by antiinflammatory drugs of the response of corneal sensory nerve fibers to chemical irritation. AB - PURPOSE: Nonsteroidal antiinflammatory drugs (NSAIDs) have been applied topically to reduce ocular pain caused by corneal injury or anterior segment surgery. The authors investigated whether the analgesic effects of the NSAIDs diclofenac, indomethacin, and flurbiprofen and of the calcium channel antagonist diltiazem on corneal pain are mediated by a reduction of nerve activity in corneal polymodal nociceptive fibers. METHODS: Impulse activity of single A-delta and C corneal nerve fibers was recorded from the ciliary nerves of anesthetized cats. Polymodal units were identified by their response to both touching with the Cochet-Bonnet esthesiometer and to acidic stimulation with 30-second pulses of 80% or 98.5% CO2 or 60 microl of 10 mM acetic acid, applied to the corneal receptive area. Ongoing impulse activity, firing responses to CO2 or acetic acid, and mechanical threshold of single fibers were recorded before and at various times (5 to 90 minutes) after topical application of 0.1% sodium diclofenac, 0.03% sodium flurbiprofen, 0.1% indomethacin, and 0.045% diltiazem hydrochloride or of their vehicles. RESULTS: Indomethacin, diclofenac, and flurbiprofen, in decreasing order of potency, gradually reduced the mean frequency of the impulse response of corneal polymodal nerve fibers evoked by CO2 stimuli. The progressive increase of ongoing activity, observed in vehicle-treated eyes after repeated CO2 stimulation was also prevented by NSAIDs. Diltiazem also attenuated the response to CO2 for a shorter period of time and with a faster time course. The mechanical threshold of corneal polymodal fibers was not affected by treatment with any of these drugs. CONCLUSIONS: Indomethacin, diclofenac, and flurbiprofen, as well as the calcium antagonist diltiazem, diminish the responsiveness of corneal polymodal nociceptors to chemical stimuli. This appears to be caused, in part, by a direct effect of these drugs on the excitability of polymodal nerve endings, but also by an inhibition by NSAIDs of the formation of cyclooxygenase products such as prostaglandins, thus reducing the enhanced responsiveness of nociceptors caused by local release of arachidonic acid metabolites from injured cells. PMID- 9331259 TI - Topical formulations of novel angiostatic steroids inhibit rabbit corneal neovascularization. AB - PURPOSE: To evaluate the antiangiogenic potential of topical ophthalmic formulations of the novel angiostatic steroids AL-3789 and AL-4940, using a rabbit model of corneal neovascularization. METHODS: Neovascularization was induced in the rabbit cornea by surgical implantation of a standard ethylene vinyl acetate copolymer (Elvax-40) pellet containing 1 microg lipopolysaccharide. Coded formulations of the control vehicle or the following test agents were administered in prevention and intervention treatment protocols: 1% formulations of AL-3789, AL-4940, and cortisol acetate as a positive drug control. Three doses of AL-3789 (0.01%, 0.1%, and 1%) were also evaluated in a prevention treatment protocol. Corneal responses were monitored throughout a 2-week treatment period, and 1 week after the last treatment dose. Observations included quantitative measurement of the area of new blood vessel growth and qualitative assessment of cellular infiltrate and edema. All treatments and observations were performed in a double-masked manner. RESULTS: All tested formulations, except the vehicle and the 0.01% AL-3789 preparation, significantly inhibited corneal neovascularization and other lipopolysaccharide-induced responses in the various treatment protocols employed. AL-4940, the free alcohol form of AL-3789, was slightly less effective than cortisol acetate or AL-3789. The extent of inhibition of the angiogenic response by the 1% and 0.1% AL-3789 suspensions ranged from 76% to 100% 1 week after the last treatment. CONCLUSIONS: The antiangiogenic steroid AL-3789 may be a therapeutically useful angiostatic agent for corneal neovascularization and potentially could be effective in other ocular neovascular diseases. PMID- 9331260 TI - Alkali burn-induced synthesis of inflammatory eicosanoids in rabbit corneal epithelium. AB - PURPOSE: Alkali burning of the rabbit cornea is a well-established model for the study of anterior surface inflammation, neovascularization, and wound-healing processes. 12-hydroxyeicosanoids have been implicated as mediators of such responses. 12(S)-hydroxyeicosatetraenoic acid (12[S]-HETE) is a lipoxygenase derived arachidonate metabolite and 12(R)-hydroxyeicosatetraenoic acid (12[R] HETE) is formed by a cytochrome P450 monooxygenase; both give rise to the potent angiogenic factor 12(R)-hydroxyeicosatrienoic acid (12[R]-HETrE). In this study, the authors correlate the pattern of their synthesis in the corneal epithelium with the inflammatory response after alkali injury. METHODS: New Zealand albino rabbits were anesthetized and alkali burns created with 10-mm filter paper discs (1 N NaOH for 2 minutes). Corneas were then rinsed; 1 to 7 days later, corneal epithelium was scraped and used to assess 14C-arachidonic acid conversion to 12 HETE and 12-HETrE enantiomers in the presence of NADPH by chiral high-pressure liquid chromatography. The inflammatory response secondary to the alkali burn was quantified through area measurements of reepithelialization and neovascularization. RESULTS: Alkali burn induced a time-dependent production of corneal epithelial 12-HETE and 12-HETrE. A marked increase in 12-HETE and 12 HETrE synthesis was evident at day 2 (from 22 +/- 7 to 139 +/- 22 ng/hour) after injury, increasing to 800 +/- 68 ng/hour at day 7. Chiral analysis revealed a time-dependent synthesis of the R and S enantiomers of 12-HETE (24% R, 76% S) and 12-HETrE (72% R, 28% S). Total arachidonate metabolism, as well as the formation of 12(R)-HETrE, correlated with the area of neovascularization (P < 0.01 and P < 0.02, respectively). CONCLUSIONS: The results demonstrate that surviving and regenerating epithelium has an increased capacity of synthesizing 12(S)-HETE and 12(R)-HETE and that maximal production of 12(R)-HETrE, a known direct and indirect angiogenic factor, coincides with neovascularization in this model. Thus, the lipoxygenase and cytochrome P450-dependent activities increased in a time-dependent manner, indicating the potential involvement of both pathways in the inflammatory response to alkali burn. The formation of significant quantities of 12(R)-HETE and 12(R)-HETrE is a novel finding in this alkali injury model. PMID- 9331261 TI - Dominant and digenic mutations in the peripherin/RDS and ROM1 genes in retinitis pigmentosa. AB - PURPOSE: To measure the proportion of cases of retinitis pigmentosa (RP) caused by mutations in the peripherin/RDS (RDS) and ROM1 genes. METHODS: The single strand conformation polymorphism (SSCP) method was used to analyze 227 unrelated patients with dominant or recessive RP for mutations in the RDS gene and an overlapping set of 315 unrelated patients for mutations in the ROM1 gene (excluding patients with other known RP genes). Variant bands revealed by SSCP were studied further by polymerase chain reaction-based, direct genomic sequencing and, where possible, by cosegregation analysis in the families of the index cases. RESULTS: Four index patients were found to have RP as a result of one of four dominant mutations in the RDS gene, two of which are novel. Four other index patients were found to have digenic RP as a result of the combination of heterozygous mutations in both the RDS and the ROM1 gene, with one of the ROM1 mutations being novel. The digenic cases all had the same RDS mutation (the missense change Leu185Pro), but each had one of three different ROM1 mutations. The authors were unable to determine through cosegregation analysis whether three other changes encountered in the RDS gene and five in the ROM1 gene were pathogenic. CONCLUSIONS: The authors found mutations in the RDS gene as a cause of dominant or digenic RP and mutations in the ROM1 gene as a cause of digenic RP. No cases of RP caused by ROM1 mutations alone have been discovered thus far. Mutations in the RDS and ROM1 genes are infrequent causes of RP, together accounting for only a few percent of patients in the United States and Canada. PMID- 9331262 TI - Disease expression in X-linked retinitis pigmentosa caused by a putative null mutation in the RPGR gene. AB - PURPOSE: To determine the disease expression in X-linked retinitis pigmentosa (XLRP) caused by a putative null mutation in the RPGR (retinitis pigmentosa GTPase regulator) gene. METHODS: In a family with XLRP, haplotype analysis was performed with polymorphic microsatellite markers from the Xp chromosomal region, and genomic polymerase chain reaction sequencing was used to identify sequence variations in the RPGR gene. Hemizygotes and heterozygotes were evaluated clinically and with visual function tests. Optical coherence tomography (OCT) was performed on heterozygotes. Postmortem donor retinas from a heterozygote were examined by microscopy and immunocytochemistry. RESULTS: X-linked inheritance was confirmed by haplotype analysis using Xp markers. Sequence analysis of the RPGR gene identified a single base pair change, a G-->T transversion, that converts codon 52 GGA (Gly) to TGA (stop codon); the mutation segregates with the disease. A hemizygote in the third decade of life had barely measurable rod function and severely impaired cone function that diminished further over a 7-year interval. Heterozygotes varied in degree of disease expression from mild to severe. Perimetry showed loci with normal rod and cone sensitivity interspersed with loci having either equal rod and cone dysfunction or rod > cone dysfunction. Electroretinographic photoreceptor responses had equal reductions in rod and cone maximal amplitude. OCT cross sectional reflectance images of retinal regions with severe dysfunction showed reduced thickness of the retina and retinal pigment epithelium-choriocapillaris (RPE-CC) complex and increased reflections posteriorly. Regions with mild dysfunction showed similar OCT findings but with preserved retinal thickness. Retinal histopathology in a heterozygote revealed loss of photoreceptors throughout, with retention of only a few islands of cones with tiny or absent outer segments and rods lacking outer segments. CONCLUSIONS: This RPGR gene mutation, in its mildest expression in heterozygotes, causes a relatively equal disturbance of rod and cone photoreceptor function. Detectable structural change by OCT at the level of the RPE-CC can be present in patches of retina with minimal functional disturbance. More advanced disease stages in heterozygotes show greater rod than cone dysfunction, and the end stage in hemizygotes and heterozygotes is that of typical RP, with only barely detectable cone function from residual cones in a thinned retina with abnormal RPE and choriocapillaris. PMID- 9331263 TI - Single exposures to antiproliferatives: long-term effects on ocular fibroblast wound-healing behavior. AB - PURPOSE: To determine the long-term effects of single, 5-minute exposures to 5 fluorouracil (5FU) and mitomycin-C (MMC) on Tenon's capsule fibroblast migration, growth factor production, growth factor receptor expression, and extracellular matrix (ECM) production. METHODS: Monolayer cultures and the overlying growth medium of Tenon's capsule fibroblasts exposed to 5FU (0.25 to 25 mg/ml) or MMC (0.001 to 0.1 mg/ml) were harvested up to 48 days after treatment. The expression of growth factors and growth factor receptors, including transforming growth factor beta (TGFbeta), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF), and ECM molecules (collagen type I, collagen type III, and fibronectin) were quantitated at the mRNA and protein levels. The ability of fibroblasts exposed to 5FU and MMC to migrate to fetal calf serum was also investigated up to 48 days after treatment. RESULTS: Control cultures were found to produce the growth factors TGFbeta and bFGF but not EGF. Exposure to 5FU or MMC resulted in an initial significant increase (P < 0.05) in the production of TGFbeta and bFGF, with levels then decreasing toward those of controls. Cells exposed to 5FU or MMC exhibited an initial significant decrease (P < 0.05) in the number of TGFbeta, bFGF, and EGF growth factor receptors, with subsequent recovery toward control levels by day 48 after treatment. Both 5FU and MMC caused a significant reduction (P < 0.05) in collagen type I and fibronectin production compared to controls throughout the 48-day culture period. The production of collagen type III was initially elevated (P < 0.05) compared to controls after exposure to 5FU or MMC, production then decreasing toward control levels over the remainder of the 48-day culture period. The migration of cells exposed to 5FU or MMC was significantly reduced (P < 0.05) compared to controls up to 48 days after treatment; these cells exhibited a partial recovery of migratory ability throughout this period. CONCLUSIONS: Fibroblasts whose growth was arrested using single, short exposures to 5FU or MMC appear to be capable of performing several crucial aspects of wound healing, including the expression of growth factors and receptors and ECM molecules and the ability to migrate. These findings may help explain why in some patients treated with antiproliferatives, glaucoma filtration surgery fails because of scarring. PMID- 9331264 TI - Autopsy analysis of clinically unilateral exfoliation syndrome. AB - PURPOSE: To study the pathogenesis of clinically unilateral exfoliation syndrome by localizing exfoliation deposits in involved and fellow eyes during autopsy. METHODS: The formalin-fixed and paraffin-embedded involved and fellow eyes were obtained at autopsy from five patients (age range, 72 to 88 years) with clinically unilateral exfoliation. Exfoliation deposits were identified with monoclonal antibodies (mAb) HNK-1 and NC-1 to the HNK-1 carbohydrate epitope, and with five lectins (Bauhinia purpurea agglutinin, Concanavalin A, Lens culinaris agglutinin, Phaseolus vulgaris erythroagglutinin, and Ricinus communis agglutinin I) using the avidin-biotinylated peroxidase complex (ABC) method. RESULTS: Marked exfoliation deposits in all involved eyes, and weak exfoliation deposits in one fellow eye were consistently detected in light microscopic, immunohistochemical, and lectin histochemical examinations. Similarly labeled deposits were present around a population of blood vessels of the iris in every involved and fellow eye. Particularly in fellow eyes, these subendothelial deposits were better visualized with mAbs to the HNK-1 epitope than they were with lectins. In the only fellow eye with early exfoliation, the reactivity around blood vessels was more conspicuous than the exfoliation deposits, whereas the reverse was true in the involved eyes. CONCLUSIONS: Clinically unilateral exfoliation is asymmetric, rather than truly monocular. The findings in fellow eyes suggest that iris blood vessels become abnormal early in the process, even before exfoliation deposits are histopathologically seen in the posterior chamber. Marked asymmetry in exfoliation indicates an influence of modulating local factors that may be internal or external to the eye, and that also may be functional in bilateral exfoliation. PMID- 9331265 TI - Development and aging of cell topography in the human retinal pigment epithelium. AB - PURPOSE: To determine how regional cell density of this tissue changes with age, the authors examined the topography of the human retinal pigment epithelium (RPE) in wholemounted tissue obtained from eyes aged 12 to 89 years, donated for corneas. METHODS: The RPE, with choroid attached, was wholemounted and stained with cresyl violet. From these preparations, the authors analyzed retinal area, RPE cell number, and cell density. RESULTS: Retinal pigment epithelial cell number is highly variable between persons but does not appear to be age related. Retinal area increases until approximately 30 years of age, but beyond this age individual variation masks further enlargement. The distinctive topography of the RPE changes markedly with age. There is a modification from the relatively homogeneous cell density distribution in the youngest retinas examined toward a more heterogeneous pattern in older retinas. From mid-adolescence, a band of larger cells appears at the extreme periphery, adjacent to the ora serrata, which gradually widens so that by 90 years of age, it occupies the outermost 30% of the retinal area. Cell density is highest in the central temporal retina, adjacent to the macula in the neural retina, throughout life. Cell density values in this region increase slightly with age, and the difference between this and surrounding regions becomes more marked with age. CONCLUSIONS: With no marked change in total cell number, peripheral RPE in humans enlarges in area throughout life, but the RPE in more central regions decreases in area. PMID- 9331266 TI - Expression of CD44 and variant isoforms in cultured human retinal pigment epithelial cells. AB - PURPOSE: CD44 is a major hyaluronic acid receptor that exists as a number of isoforms, generated by alternative splicing of 9 "variant" exons in humans (v2 to v10) and 10 exons in rodents. Little is known about the expression and function of CD44 in human retinal pigment epithelium (RPE) cells. Therefore, the authors determined whether human RPE cells express CD44, and whether the expression differs depending on the proliferative status of the cells. METHODS: Human RPE cells were harvested from normal donor eyes and propagated in culture. Total RNA was extracted from cultured cells. mRNA expression of CD44 was determined by reverse transcription-polymerase chain reaction (RT-PCR), followed by cloning and sequencing of the PCR products, and by Southern hybridization. CD44 cell surface expression was measured by flow cytometry. Western hybridization and immunohistochemistry were used to determine the CD44 immunoreactivity of cultured human RPE cells and normal human RPE cells in situ. RESULTS: The standard form of CD44 mRNA and variant isoforms containing exon v6 or v10 were expressed in cultured human RPE cells. CD44 mRNA and protein levels were increased in proliferating human RPE cells compared with density-arrested counterparts. Addition of 1 microM retinoic acid enhanced the cell density-induced downregulation of CD44 mRNA, but did not significantly affect the CD44 cell surface protein expression. As previously reported, CD44 immunoreactivity was not detected in normal human RPE cells in situ. CONCLUSIONS: Cultured human RPE cells express CD44 standard form and variant isoforms containing exon v6 or v10, which are preferentially expressed by proliferating human RPE cells. PMID- 9331267 TI - The role of neuronal and endothelial nitric oxide synthase in retinal excitotoxicity. AB - PURPOSE: Nitric oxide synthase (NOS) plays an essential role in neuronal function and is critical in the brain for normal and pathologic responses to glutamate. The role of NOS in the retina is less well understood. The retina provides an experimental system in which the intrinsic circuitry is well defined; retinal excitotoxic damage has been well characterized. METHODS: To determine whether neuronal NOS (nNOS) and endothelial NOS (eNOS) are critical in excitotoxic damage in the retina, nNOS- and eNOS-deficient mice were subjected to intravitreal injections of N-methyl-D-aspartate (NMDA) or to arterial occlusions. RESULTS: Retinal ganglion cells in the nNOS-deficient mouse were relatively resistant to NMDA and to arterial occlusion. In contrast, the damage in the eNOS-deficient mouse retina was not distinguishable from that in control animals. Preinjection with an NOS inhibitor was partially protective. CONCLUSIONS: The presence of nNOS is a prerequisite for the full expression of excitotoxicity in the retina; eNOS does not appear to play a significant role. PMID- 9331268 TI - Expression of inducible nitric oxide synthase in bovine corneal endothelial cells and keratocytes in vitro after lipopolysaccharide and cytokines stimulation. AB - PURPOSE: To determine whether bovine corneal endothelial (BCE) cells and keratocytes express the inducible form of nitric oxide synthase (NOS) after exposure to cytokines and lipopolysaccharide (LPS), and to study the regulation of NOS by growth factors. METHODS: Cultures of bovine corneal endothelial cells and keratocytes were exposed to increasing concentrations of LPS, interferon gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha). At selected intervals after exposure, nitrite levels in the supernatants were evaluated by the Griess reaction. Total RNA was extracted from the cell cultures, and messenger RNA levels for inducible NOS (NOS-2) were measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Exposure of BCE cells and keratocytes to LPS and IFN-gamma resulted in an increase of nitrite levels that was potentiate by the addition of TNF-alpha. Analysis by RT-PCR demonstrated that nitrite release was correlated to the expression of NOS-2 messenger RNA in BCE cells and keratocytes. Stereoselective inhibitors of NOS and cycloheximide inhibited LPS-IFN-gamma-induced nitrite release in both cells, whereas transforming growth factor-beta (TGF-beta) slightly potentiated it. Fibroblast growth factor-2 (FGF-2) inhibited LPS-IFN-gamma-induced nitrite release and NOS-2 messenger RNA accumulation in keratocytes but not in BCE cells. CONCLUSIONS: The results demonstrate that in vitro activation of keratocytes and BCE cells by LPS and cytokines induces NOS-2 expression and release of large amounts of NO. The high amounts of NO could be involved in inflammatory corneal diseases in vivo. PMID- 9331269 TI - Tractional force generation by porcine Muller cells: stimulation by growth factors in human vitreous. AB - PURPOSE: To examine the levels of Muller cell contraction-stimulating activity in human vitreous, correlate these levels with clinical presentation, and identify, the causative growth factors. METHODS: Human vitreous was collected from patients undergoing pars plana vitrectomy (n = 84). Muller cells were isolated from porcine retina and maintained in tissue culture. Tractional forces generated by cells incubated on three-dimensional collagen gels were measured as changes in gel thickness. Contraction-stimulating activity in vitreous (VA) was calculated from the close-response profiles of gel contraction to vitreous protein. The contributions of individual growth factors to vitreous activity (n = 10) were assessed by inhibition with specific neutralizing antibodies. RESULTS: The mean VA of patients with retinal detachment (3.65) and proliferative vitreoretinopathy stages A, B, and C (2.06) were elevated above that of patients without retinal pathology (vitreous activity = 0.23) or retinal defects alone (0.57). Mean activities in patients with epimacular proliferation (1.22) and vitreous hemorrhage (1.40) were also significantly elevated. The percentage of this activity attributable to insulin-like growth factor 1 (IGF-1) varied from 9.2% to 84.5% with a mean of 61.3%. Similarly, the percent contribution of platelet derived growth factor (PDGF) ranged from 6.8% to 49.0% with a mean of 26.5%. CONCLUSIONS: The vitreous of patients with retinal detachment, proliferative retinal disease, and vitreous hemorrhage contain varying amounts of growth factors that stimulate tractional force generation by Muller cells. The majority of the activity can be attributed to IGF-1 and a smaller proportion to PDGF. PMID- 9331270 TI - Cell attachment to, and contraction of, the retina in vitro. AB - PURPOSE: To examine the behavior of fibroblasts and retinal pigment epithelial cells after attachment to the retinal surface in vitro to elucidate the pathobiology of the early stages of epiretinal membrane formation. METHODS: Human retinal pigment epithelial (HRPE) cells and bovine Tenon's capsule fibroblasts (BTFs) were seeded onto the surface of bovine retinal explants maintained in organ culture. The changes induced in the underlying retina, including contraction, were assessed during a period of up to 10 days. Immunohistochemistry was used to assess proliferation of the seeded cells and to determine deposition of extracellular matrix. RESULTS: Explants of bovine neuroretina were maintained in organ culture, with good morphologic preservation of the inner limiting lamina and inner retinal layers, for 7 to 10 days. The HRPE cells and the BTFs attached to the retinal surface and exerted tractional forces, producing partial- and full thickness retinal folding. Contraction commenced within 24 hours of attachment of the cells and continued for several days, with most of the contraction occurring within the next 48 to 72 hours. The HRPE cells and BTFs were found to be equally contractile. Deposition of cellular fibronectin (but not collagen type I) was demonstrated. CONCLUSIONS: The contractile cellular membranes generated in this organ culture system exhibit many of the morphologic and functional features of epiretinal membranes found in the early stages of proliferative vitreoretinopathy. PMID- 9331271 TI - Expression of fibroblast growth factor-5 by bovine choroidal endothelial cells in vitro. AB - PURPOSE: To demonstrate the expression of fibroblast growth factor-5 (FGF-5) by bovine choroidal microvascular endothelial (BCME) cells and to investigate its possible role as an autocrine mitogen in these cells. METHODS: Expression of FGF 5 by BCME cells was studied by a combination of Northern and Western blot analyses. Total RNA was isolated from BCME cultures at passages 5 through 8 and analyzed by Northern blot analysis for the presence of FGF-5 transcripts, using a 1-kb human complemetary DNA. Slot-blot analysis was performed to determine possible cross-reactivity between this probe and acidic and basic FGFs of human and bovine species. A previously characterized antibody directed against the aminoterminus of the human FGF-5 sequence was used in Western blot analyses to identify immunoreactive proteins released by BCME cells into the medium. Finally, the mitogenic activity of human recombinant FGF-5 on a variety of cell types was evaluated, using a cellular proliferation assay. RESULTS: Northern blot analysis provided evidence for the expression of two major FGF-5 transcripts at 4 kb and 3 kb and two minor transcripts at 2.2 kb and 1.7 kb. A single immunoreactive protein with a molecular weight of 34 kDa was identified by Western blot analysis of conditioned media. In cellular proliferation assays, human recombinant FGF-5 was not mitogenic in BCME cells but exhibited an approximate ED50 of 1.8 to 3.7 nM in BALB/c3T3 fibroblasts. This ED50 was within the range reported by the manufacturer, using a thymidine incorporation assay and a similar embryonic fibroblast cell line. Fibroblast growth factor-5 also stimulated proliferation of human retinal pigment epithelial cells. CONCLUSIONS: Bovine choroidal microvascular endothelial cells exhibit expression in vitro of FGF-5 at the messenger RNA and protein levels. Perivascular and endothelial cell staining for FGF-5 seen previously in choroidal neovascular membranes may therefore arise from expression by choroidal endothelial cells. Because nonglycosylated recombinant FGF-5 does not appear to be a mitogen in BCME cells in vitro, it is reasonable to question its role as an autocrine mitogen in vivo. Fibroblast growth factor-5 may instead be serving paracrine roles in the stimulation of fibroblasts and retinal pigment epithelial cells during the formation of choroidal neovascular membranes. Studies with fully glycosylated recombinant FGF-5 will be required, however, to assess the biologic activity of this member of the FGF gene family. PMID- 9331272 TI - A histologic study (including DNA quantification and Ki-67 labeling index) in uveal melanomas after brachytherapy with ruthenium plaques. AB - PURPOSE: To investigate the proliferative potential and DNA damage in uveal melanomas treated by brachytherapy. METHODS: Forty-two enucleated eyes that had been treated with 106Ru/106Rh radioactive plaques for uveal melanoma were subgrouped according to the extent of irradiation damage. Cell proliferation was determined by immunoreactivity for the proliferation marker Ki-67 (Mib-1) and ploidy by quantitative DNA image analysis. Thirty globes containing uveal melanomas without prior brachytherapy served as a control group. RESULTS: The values for Ki-67 reactivity and ploidy could be correlated with radiation-induced changes within the tumors. In regions of the tumor where complete exposure to the prescribed radiation dose was assumed from the histologic findings, the Ki-67 index was close to or equal to zero. Hypoploidy was exclusive to irradiated tumors and was most often detected in effectively irradiated regions. Tumor regions classified as partially irradiated or recurrent showed an increase of Ki 67 indices and DNA content. Values obtained in recurrent tumors did not significantly differ from the control group. CONCLUSIONS: Tumor cell proliferation and variations in ploidy status could be detected after brachytherapy, but the response varied markedly both within individual tumors and within the irradiated group. Evidence of persisting proliferative potential could be obtained in ostensibly sterilized tumor tissue, but a negligible Ki-67 index and the presence of hypoploidy were considered to be reliable indicators for radiation-induced loss of proliferative potential. PMID- 9331274 TI - Stability of ocular counterrolling and Listing's plane during static roll-tilts. AB - PURPOSE: To investigate whether habituation occurs in ocular counterrolling (OCR), how stable shifted Listing's plane is, and what effects visual stimulation and alertness exert on the OCR and on Listing's plane. METHODS: Two monkeys (Macaca fuscata) were engaged in this experiment. A dual scleral search coil method was used for recording three-dimensional eye movements. Spontaneous eye movements were recorded for 2 hours in dark and light, while each monkey was held in different static roll positions (up to +/-34 degrees) with its head fixed. Eye movements were also recorded during the monkeys' drowsy periods in the dark. RESULTS: In alert conditions, OCR gains showed some fluctuations but did not change consistently for 2 hours in seven of seven sessions in the dark and in seven of nine sessions in the light. The OCR gains in the light did not differ from those in the dark. The thickness of shifted Listing's plane during the static roll-tilt was also stable for 2 hours and was within the range of the thickness of Listing's plane in the upright position in the light. During drowsy periods, the thickness of Listing's plane increased, and the gain in OCR decreased. CONCLUSION: Ocular counterrolling shows no habituation for 2 hours of static roll-tilts in the alert monkey. In the same condition, Listing's plane consistently maintains its precision. Visual input does not affect OCR, but alertness is necessary to keep the stability of OCR and Listing's plane. PMID- 9331273 TI - Membrane-associated carbonic anhydrase in cultured rabbit nonpigmented ciliary epithelium. AB - PURPOSE: To measure the activity of membrane-associated carbonic anhydrase (CA) in cultured rabbit nonpigmented epithelial (NPE) cells, determine its identity and its sensitivity to extracellular trypsin, and compare the ability of acetazolamide and a cell-impermeant dextran-bound CA inhibitor to change cytoplasmic pH. METHODS: Studies were conducted using a cell line derived from rabbit NPE. The cells were lysed and separated into soluble and insoluble fractions by differential centrifugation. CA activity in these fractions was determined using a CO2 hydration assay. In studies with intact cells, a membrane impermeable high-molecular-weight dextran-bound inhibitor (DBI) was synthesized and used to selectively bind and inhibit the extracellular-facing membrane-bound CA. Measurements of CA activity in intact red blood cells were conducted to confirm DBI remains extracellular. Acetazolamide, a membrane-permeable CA inhibitor, was used to inhibit total CA activity. Intracellular pH was determined using the pH-dependent absorbance of the fluorescent dye BCECF-AM. RESULTS: A low speed pellet enriched with plasma membrane material accounted for 22.3 +/- 6.1% (n = 18) of the total CA activity in the cultured NPE. When intact cells were exposed to trypsin-EDTA, a 28% reduction of membrane-associated CA activity was observed; DBI inhibited this CA activity loss. Cytosolic CA activity was inhibited by 0.2% sodium dodecyl sulfate (SDS). In contrast, membrane-associated CA was SDS resistant, a characteristic of the CA-IV isozyme. By Western blot, CA IV immunoreactive polypeptide was detected in the cultured cells and also in native rabbit and porcine ciliary epithelium. Inhibition of total CA activity with acetazolamide and inhibition of extracellular-facing membrane-associated CA with DBI caused an identical intracellular pH decrease in intact NPE cells. CONCLUSIONS: Expression of the CA-IV isozyme could account for the significant fraction of CA activity in the cultured NPE, which is membrane associated and SDS resistant. Sensitivity to tryptic hydrolysis suggests the membrane-associated CA partially faces extracellularly. As judged by responses to an extracellular CA inhibitor, the membrane-associated CA has a functional role in maintaining cytoplasmic pH. PMID- 9331275 TI - Decreased retinal ganglion cell number and misdirected axon growth associated with fissure defects in Bst/+ mutant mice. AB - PURPOSE: The autosomal semidominant mutation Bst (belly spot and tail) is often associated with small and atrophic optic nerves in adult mice and shares several important attributes with heritable optic nerve atrophy in humans. In this article, the authors present adult and developmental studies on the retinal phenotype in Bst/+ mice. METHODS: Retinal ganglion cells in adult Bst/+ mice were labeled retrogradely with horseradish peroxidase injected into the right optic tract. Labeled ganglion cells were mapped in whole-mounted retinas ipsilateral and contralateral to the injection site. The number of axons in optic nerves of these and other cases were quantified using an electron microscopic method. Eyes of neonatal, embryonic day 15 (E15), and embryonic day 12 (E12) Bst/+ mutants were examined histologically to understand the etiology of the retinal phenotype. RESULTS: Approximately 60% of adult Bst/+ mice have deficient direct pupillary light responses. This neurologic phenotype is associated with a reduction in the number of retinal ganglion cells from the wild-type average of 67,000 to less than 20,000 in Bst/+ mutants. Ganglion cells with crossed projections are more severely affected than those with uncrossed projections. Histologic analysis of eyes from E12 mice reveals a delayed closure of the optic fissure. Despite this abnormality, other ocular structures appear relatively normal. However, some E15 mutants exhibit marked disorganization of the retinal neuroepithelium, and ganglion cell axons are found between pigmented and neural retina. At birth, optic nerves of affected mice are smaller than those of wild-type mice, ectopic axons are found within the eyes, and the ganglion cell layer contains many dying cells. CONCLUSIONS: The expression of the retinal phenotype in Bst/+ mutants is highly variable-ranging from a complete absence of ganglion cells to numbers comparable to that in wild-type mice. The reduction in ganglion cell number in affected adult Bst/+ mice is attributable to the failure of ganglion cell axons to reach the optic nerve head early in development. Delayed fusion of the fissure is consistently associated with the Bst/+ genotype and probably contributes to the failure of ganglion cell axons to grow out of the eye. PMID- 9331276 TI - Modulation of major histocompatibility complex class II expression in retinas with age-related macular degeneration. AB - PURPOSE: To investigate antigenic and morphologic features of microglial and vascular elements in the neural retina associated with age-related macular degeneration (ARMD) compared with those features in age-matched normal and young adult retinas. METHODS: Adult eyes (n = 97) were classified histopathologically into normal and ARMD-associated groups. Peroxidase imunohistochemical examination of retinal flatmounts was used to visualize major histocompatibility complex class II (MHC-II) immunoreactivity; the intensity and distribution of labeling were quantified by image analysis. In histochemical investigation, reduced nicotinamide-adenine dinucleotide phosphate diaphorase and glial fibrillary acidic protein or MHC-II double labeling were used to detect vascular changes in some preparations. RESULTS: An increase in the proportion of the retina (percentage of total area) expressing MHC-II immunoreactivity was observed in age matched retinas compared with that seen in normal retinas. A significant increase (P < 0.05) in the percentage of area immunoreactive for MHC-II was observed, primarily on vascular elements, in retinas with incipient ARMD compared with the area affected in the age-matched group. Increased MHC-II immunoreactivity on vessels in the normal-aged group observed with confocal microscopy was associated with irregularities in the organization of astrocytes. Hypertrophy of retinal microglia was also apparent, although the intensity of microglial MHC-II immunoreactivity was not significantly different between groups. CONCLUSIONS: The results indicate that an increase in MHC-II immunoreactivity on retinal vascular elements is associated with normal aging. A further increase in MHC-II immunoreactivity on vascular elements and morphologic changes in microglia was associated with incipient ARMD. Immunologic responses in neural retinal microglia and vascular elements appear to be related to early pathogenetic changes in retinal pigment epithelium pigmentation and drusen formation. PMID- 9331277 TI - Effects of refractive error on detection acuity and resolution acuity in peripheral vision. AB - PURPOSE: To evaluate the effect of refractive error on detection acuity and resolution acuity in peripheral vision. METHODS: Detection acuity, defined as the highest spatial frequency for which luminance gratings can be discriminated from a uniform field, and resolution acuity, defined as the highest spatial frequency for which spatial patterns are perceived veridically, was determined for vertical and horizontal gratings located at 20 degrees, 30 degrees, and 40 degrees of eccentricity. Resolution was also measured for tumbling-E discrimination at these locations. Refractive state of the eye for test targets was manipulated by introducing an ophthalmic trial lens into the line of sight for the stimulus while holding accommodative state fixed. RESULTS: Detection acuity in the periphery varied significantly with the amount of optical defocus, whereas acuity for grating resolution or letter discrimination was unaffected by defocus over a large range (up to 6 D). These results are consistent with the working hypothesis that detection acuity in the periphery is limited by contrast insufficiency under normal viewing conditions, but resolution is limited by ambiguity because of neural undersampling. CONCLUSIONS: The large depth of focus for resolution acuity measured for peripheral vision indicates that spatial resolution is likely to remain sampling-limited even when peripheral refractive errors are not fully corrected, thus relaxing the methodologic requirements for obtaining noninvasive estimates of neural sampling density of the living eye in a clinical setting. PMID- 9331278 TI - Endothelin expression in ocular tissues of diabetic and insulin-treated rats. AB - PURPOSE: The endothelins are potent vasoactive peptides that are widely distributed in ocular tissues. There is evidence linking the endothelins to vascular dysfunction in diabetic microangiopathy. Thus, the synthesis and distribution of endothelin-1 (ET-1) and endothelin-3 (ET-3) were studied in the retinas of diabetic and nondiabetic animals. METHODS: Levels of ET-1 and ET-3 were determined by radioimmunoassay in ocular tissues of normal rats, and in rats with streptozotocin-induced diabetes of 6 and 12 weeks' duration, insulin-treated and untreated. In a separate cohort of similarly treated animals, retinal vascular trypsin digest preparations were immunostained, using antibodies raised against ET-1 and ET-3. RESULTS: Ocular ET-1 levels were elevated twofold in diabetic animals that received insulin treatment for 7 days when compared with levels in normal rats. Insulin treatment for 10 days before death caused a fourfold elevation of ET-1 in ocular tissues. Endothelin-1 was also increased in 12-week-old diabetic animals and in those maintained on insulin throughout their period of diabetes. Immunofluorescence to anti-ET-1 within the capillary bed and veins of the retina in diabetic insulin-treated animals was elevated when compared with digests from normal litter-matched control animals. Ocular tissue ET-3 levels were unaffected by diabetes. CONCLUSIONS: Overall ocular and retinal tissue levels of ET-1 were selectively elevated by diabetes and insulin treatment, suggesting that the endothelins may be involved in the pathogenesis of diabetic retinal microangiopathy. PMID- 9331279 TI - Effect of cyclosporine on anterior chamber-associated immune deviation with retinal transplantation. AB - PURPOSE: To determine whether immunosuppression using cyclosporine interferes with anterior chamber associated immune deviation (ACAID) and can promote survival of retinal allografts in the anterior chamber. METHODS: Neonatal neural retinas of C57BL/6 mice or ovalbumin were injected into the anterior chamber of BALB/c adult mice. In the test group recipients were injected with cyclosporine (10 mg/kg per day) from day 0 to 11 or from day 11 to 34 after implantation. At 12 and 35 days after transplantation, lymphocytes from the test group were injected into naive BALB/c mice to assay for the presence of suppressor T cells (adoptive transfer). The fate of the retinal grafts was determined by histologic examination at day 12 and 35. To evaluate the potential neurotoxic effects of cyclosporine in the absence of immune rejection mechanisms, cyclosporine was given to SCID mice during days 11 to 34 after syngeneic neonatal neural retinal grafts were placed in the anterior chamber. RESULTS: At 12 days after transplantation, spleens of both cyclosporine-treated and control mice contained suppressor cells against donor alloantigens. The retinal grafts in the anterior chamber of both groups of mice were fully developed and well differentiated. The same duration of administration of cyclosporine did not interfere with the production of efferent suppressor cells after inoculation of ovalbumin into the anterior chamber. At 35 days after transplantation, only spleen cells from the cyclosporine-treated group showed the capacity to suppress donor-specific delayed hypersensitivity. However, allografts in the cyclosporine group had deteriorated by 35 days in a fashion similar to the control group. Syngeneic grafts in SCID mice showed differentiated retinal layers 35 days after transplantation. CONCLUSIONS: Cyclosporine treatment does not interfere with the ability of allogeneic neonatal retinal grafts to induce anterior chamber associated immune deviation when placed in the anterior chamber, nor does prolonged treatment with this drug interfere with the persistence of allospecific suppressor cells for 35 days after the graft. Because 35-day grafts of cyclosporine-treated mice display histologic evidence of graft failure similar to grafts placed in the anterior chamber of untreated mice, graft destruction is either the result of immune effector mechanisms not inhibited by cyclosporine, or the consequence of nonimmunologic factors. PMID- 9331280 TI - Increased plasma levels of substance P in vernal keratoconjunctivitis. AB - PURPOSE: The increase of nerve growth factor (NGF) plasma levels in vernal keratoconjunctivitis (VKC) patients has been demonstrated previously. Results of numerous studies in vitro and in vivo have shown that NGF modulates the synthesis of substance P (SP), a neuropeptide involved in the pathogenesis of human allergic diseases. In this study the involvement of SP in this allergic conjunctivitis is investigated, along with its relationship with NGF and other systemic and local markers of VKC. METHODS: Competitive radioimmunoassays were used to detect the levels of SP in plasma, the levels of eosinophil cationic protein, and the total and specific immunoglobulin E in the serum of 11 patients with VKC and in 11 healthy matched controls. Plasma levels of nerve growth factor (NGF) were measured in all VKC patients and controls using an immunoenzymatic assay. Histologic evaluation was performed in tarsal and bulbar conjunctival specimens obtained in biopsies from 8 VKC patients and 4 control subjects. RESULTS: Patients with VKC show a significant increase of SP and NGF plasma levels (P < 0.003 and P < 0.001, respectively), and an increase of eosinophil cationic protein and immunoglobulin E levels in the serum (P < 0.001 and P < 0.002, respectively). Mast cells, eosinophils, and lymphocytes were also significantly increased in the conjunctiva of VKC patients. Interestingly enough, VKC patients with the highest NGF plasma levels also showed the highest SP levels. CONCLUSIONS: The data show the involvement of SP in VKC and suggest that SP with NGF could modulate the allergic response in this disease, probably through an interaction with inflammatory cytokines. PMID- 9331281 TI - Effects of time of storage, albumin, and osmolality changes on outflow facility (C) of bovine anterior segment in vitro. AB - PURPOSE: To analyze the influence of time of storage, the presence of albumin at physiological concentrations, and the perfusion with anisosmotic media on the aqueous humor outflow facility (C) of isolated bovine anterior segments (AS). METHODS: Anterior segments dissected from cow eyes were perfused at a constant pressure of 10 mm Hg with Dulbecco's modified Eagle's medium (DMEM; osmolality 300 mOsm/kg), with hyposmotic media (150, 210, and 270 mOsm/kg), or with hyperosmotic media (360, 420, and 480 mOsm/kg). Outflow facility was calculated every 5 seconds as the ratio between average inflow from the reservoir (in microliters per minute) and the perfusion pressure (in millimeters of mercury). Three groups were studied: a 0-hour group, with AS perfused with DMEM 1 to 3 hours after enucleation; a 0-hour alb-group, with AS perfused with DMEM plus 0.1 mg/ml albumin 1 to 3 hours after enucleation; and a 24-hour group, with AS perfused after storage for 24 hours in DMEM. In the 0-hour groups, perfusion with increasingly hyposmotic or hyperosmotic media was also made in 30-minute steps, followed by a return to isosmotic medium for 90 minutes. RESULTS: Perfusion of AS with DMEM for 9 hours caused a progressive increase in C that was statistically significant at 225 minutes in the 0-hour group perfused with DMEM and at 195 minutes in the 24-hour group perfused with DMEM. The 0-hour alb-group perfused with DMEM did not show changes in C throughout the 9-hour perfusion period. Perfusion with increasingly hyposmotic media induced a progressive decrease in C that did not recover on return to isotonic medium. Hyperosmotic media caused a progressive increase in C that returned to control values when isotonic medium was again perfused. CONCLUSIONS: Preservation of tissue for C measurements is best achieved with short storage time (1 to 3 hours). Physiological concentrations of albumin (0.1 mg/ml) prevent development of washout, suggesting that albumin or an albumin-bound factor in aqueous humor may play a role in the maintenance of outflow resistance. Outflow facility also may be influenced by volume changes in the trabecular meshwork. PMID- 9331282 TI - Induction of experimental autoimmune anterior uveitis by a self-antigen: melanin complex without adjuvant. AB - PURPOSE: Experimental autoimmune anterior uveitis (EAAU) is an organ-specific autoimmune disease induced by immunization with bovine melanin-associated antigen (MAA) and two adjuvants (complete Freund's adjuvant and purified pertussis toxin). This study was undertaken to explore whether an adjuvant is required in the induction of EAAU. METHODS: Insoluble MAA was extracted from the bovine iris and ciliary body. Soluble bovine MAA was derived by treatment of insoluble MAA with the proteolytic enzyme, V8 protease. Lewis rats were immunized with the insoluble or soluble antigen, with or without adjuvant (complete Freund's adjuvant and purified pertussis toxin). Adoptive transfer of CD4+ and CD8+ T cells was performed to investigate the pathogenesis of EAAU. RESULTS: Experimental autoimmune anterior uveitis can be induced in Lewis rats by immunization with 100 g insoluble bovine MAA alone without the use of adjuvants. The disease can be adoptively transferred to naive syngenic rats by primed CD4+ T cells. In contrast, soluble bovine MAA was not uveitogenic unless adjuvants were employed. CONCLUSIONS: The data suggest that EAAU can be induced in the Lewis rat without addition of an adjuvant. Future studies concerning the pathogenesis of EAAU can now be performed without the possible confounding effect of an adjuvant. PMID- 9331284 TI - Lipoblastoma/lipoblastomatosis: a clinicopathologic study of 25 tumors. AB - Lipoblastoma/lipoblastomatosis is an uncommon benign adipose tissue tumor of children. Since 1958, 25 of these tumors from 24 patients have been reviewed in the Department of Pathology at The Children's Hospital of Philadelphia. Tumors were resected from 19 boys (79%) and five girls, and 20 patients (84%) were < or =5 years of age at diagnosis. Twenty-three tumors presented as painless superficial soft-tissue masses; one tumor was retroperitoneal and was discovered because of vomiting; one hand tumor was present at birth. Tumors occurred in an extremity (n = 11 patients), the head and neck (n = 5), groin (n = 2), axilla (n = 2), back (n = 1), chest (n = 1), flank (n = 1), labia (n = 1), and retroperitoneum (n = 1). Thirteen tumors occurred on the left side, and five occurred on the right. Lesions measured 1.0-21.0 cm in greatest dimension; 15 of 25 (60%) measured < or =5.0 cm. The largest (retroperitoneal) tumor weighed 450 g. Eleven tumors were discrete lipoblastoma, and 14 had irregular margins (lipoblastomatosis). Microscopically, the tumors displayed adipocytes in different stages of maturation; lobules bordered by septae that were cellular in 11 cases; prominent blood vessels in 19 cases; and myxoid foci in 13 cases. Chart review of 22 patients showed that one tumor recurred 4 years after resection; one tumor recurred after 7 years as fibrolipoma; and one incompletely resected tumor enlarged and at second resection was lipoma. There were no metastases. Three patients also had hemangioma. Juvenile aponeurotic fibroma occurred in one patient near the site of resection of a lipoblastoma 4 years earlier. We conclude that lipoblastoma/lipoblastomatosis behaves benignly, occurs in both superficial and deep sites, occasionally attains large size, may mature, can recur, and may be associated with other benign soft-tissue lesions. Complete surgical excision is the treatment of choice. PMID- 9331283 TI - Epithelioid angiomyolipoma of the kidney: a report of five cases with a prominent and diagnostically confusing epithelioid smooth muscle component. AB - We report five angiomyolipomas with a prominent component of epithelioid smooth muscle cells that occurred in patients from 20 to 48 (mean, 36) years of age. The tumors often posed problems in diagnosis, particularly with regard to distinction from renal cell carcinoma. Two patients had tuberous sclerosis. Two patients with more than 5 years' follow-up are alive and well. The epithelioid smooth muscle cells typically formed sheets that in two tumors were traversed by hyaline cords. The epithelioid cells ranged from medium sized and polygonal with slightly pleomorphic nuclei and eosinophilic cytoplasm to giant cells with prominent nucleoli. Hemorrhage, necrosis, and clusters of foamy macrophages were present in three tumors. Mitotic figures were easy to find in two of the tumors, but they were absent in the others. Obvious elements of typical angiomyolipoma were present in two tumors. The others contained only scattered, thick-walled blood vessels or a few fat cells suggestive of typical angiomyolipoma. None of the tumors was positive for low- or high-molecular-weight cytokeratins or epithelial membrane antigen. Actin was detected in the epithelioid areas in four tumors. Melanoma-associated antigens related to the gp100-cl gene product, HMB-45 and HMB 50, were present in all the tumors. Another melanoma-associated antigen, CD63 (NKI/C3), also was present in all the tumors, a finding suggesting that angiomyolipoma has features in common with melanoma beyond premelanosomal structures. PMID- 9331285 TI - Intravascularly disseminated angiosarcoma: true neoplastic angioendotheliomatosis? Report of two cases. AB - Although vascular invasion is common in many malignant tumors, disseminated intravascular anaplastic neoplasms with occult primary tumor are rare occurrences. Intravascular malignant lymphoma, also called angiotropic lymphoma, is a rare variant of large cell lymphoma predominantly involving vessels in multiple organs, and usually without significant nodal involvement. Although initially misinterpreted as an endothelial neoplasm-angioendotheliomatosis immunohistochemical studies subsequently proved it to represent a peculiar form of malignant lymphoma. In this report, we describe two patients with extensive intravascular dissemination of angiosarcoma initially without clinically obvious primary tumor. These may be interpreted as examples of true angioendotheliomatosis. In each case the immunohistochemical studies ruled out the most common intravascular malignant neoplasms. The diagnosis of intravascular angiosarcoma was confirmed by the immunoreactivity of the tumor cells to several markers of endothelial lineage in both cases. Thus, angiosarcoma may present with intravascular dissemination and occult primary tumor and closely resemble metastatic carcinoma, melanoma, or angiotropic lymphoma. Immunohistochemical studies are crucial in ruling out these possibilities and in confirming the endothelial origin of the neoplastic cells. PMID- 9331286 TI - The morphologic spectrum of ovarian metastases of appendiceal adenocarcinomas: a clinicopathologic and immunohistochemical analysis of tumors often misinterpreted as primary ovarian tumors or metastatic tumors from other gastrointestinal sites. AB - Twenty cases of ovarian metastases derived from appendiceal adenocarcinomas were analyzed. The most common presentation was a pelvic mass. The appendiceal and ovarian tumors were diagnosed concurrently in 15 cases; in the remaining five, the ovarian tumors were diagnosed before the appendiceal tumor. The appendiceal adenocarcinomas demonstrated four morphologic patterns: 1) signet ring cell type, with or without glandular or goblet cell differentiation (14 cases); 2) mixed signet ring cell and intestinal type (two cases); 3) intestinal type (two cases); and 4) typical colorectal type (two cases). The ovarian tumors were bilateral in 16 cases and were histologically similar to the associated appendiceal tumor in each case. Ovarian metastases that demonstrate signet ring cell, glandular, and goblet cell differentiation mimic metastases from gastric adenocarcinoma. Those that are derived from well-differentiated mucinous appendiceal adenocarcinomas mimic primary ovarian mucinous tumors and metastases from the pancreas and biliary tract. Metastases of appendiceal adenocarcinomas of colorectal type simulate both metastatic colorectal carcinoma and primary ovarian endometrioid carcinomas. The appendiceal and ovarian tumors were immunophenotypically identical in each case. Approximately 50% of the appendiceal and ovarian tumors were positive for cytokeratin 7 (CK 7), and all were positive for cytokeratin 20 (CK 20). CK 20 positivity of the ovarian tumors is consistent with gastrointestinal origin; CK 7 positivity does not confirm ovarian origin, because appendiceal carcinomas are positive in 50% of cases. Metastatic appendiceal adenocarcinoma should be considered in the differential diagnosis of mucinous ovarian tumors with signet ring cell, goblet cell, or intestinal type differentiation, especially when these tumors are associated with extraovarian disease and are bilateral. PMID- 9331287 TI - Granulocytic sarcoma of the female genital tract: a clinicopathologic study of 11 cases. AB - Eleven patients, 13 to 76 (mean, 40) years of age, had granulocytic sarcoma of the female genital tract (FGT) (ovary, seven cases; vagina, three cases; cervix, one case). In nine cases, the FGT involvement was the initial clinical presentation of the disease, and in the other two cases, the FGT involvement was discovered during a relapse of acute myeloid leukemia. The tumors ranged from 0.5 to 14 (mean, 7.5) cm in greatest dimension. Two ovarian tumors were bilateral, and three were green. Microscopic examination revealed a predominantly diffuse pattern of growth, but cords and pseudoacinar spaces were also present focally in several cases. Sclerosis was seen in five tumors and was prominent in one. Prominent myeloid differentiation was readily recognizable on routinely stained sections in three cases, whereas the neoplastic cells in the other cases were primitive with only rare eosinophilic myelocytes. All 11 tumors were positive for chloroacetate esterase, nine of nine were strongly and diffusely positive for lysozyme, eight of eight for myeloperoxidase, seven of seven for CD68, and six of six for CD43. Examination of bone marrow or peripheral blood performed after the diagnosis of FGT involvement revealed acute myeloid leukemia in three of five cases. Two of these patients died of disease, 1 and 16 months after the initial diagnosis, and the third, who received chemotherapy, is alive and free of disease 8 months after the initial diagnosis. One of the two patients with negative bone marrow had recurrent granulocytic sarcoma 30 months after diagnosis and died of sepsis 1 month later; no residual disease was noted at autopsy. The other patient is alive and free of disease 18 months after the diagnosis. One of the four remaining patients with primary FGT involvement who did not have a bone marrow biopsy died of leukemia 24 months later; no follow-up information is available for the other three patients. One of the two patients with a prior diagnosis of acute myeloid leukemia was alive with disease 26 months later; follow-up is not available for the second patient. The diagnosis was often difficult in these cases, the most common problem being distinction from malignant lymphoma, but carcinoma, granulosa cell tumor, and, rarely, other tumors were considered. Immunohistochemical and enzyme histochemical staining were useful in establishing the diagnosis, although suspicion of the diagnosis on examination of routinely stained sections was of paramount importance. PMID- 9331288 TI - Inflammatory myofibroblastic tumor of bone: report of two cases with evidence of clonal chromosomal changes. AB - Inflammatory myofibroblastic tumor (inflammatory pseudotumor) is a pseudosarcomatous lesion that is recognized with increasing frequency in various anatomic locations. However, this lesion has not been previously reported in bone. We report on two cases of inflammatory myofibroblastic tumor occurring in bone in young adults. Both tumors presented as slightly painful, osteolytic, and well-delineated lesions of the distal femur, with a hyperintense signal on T2 weighted magnetic resonance imaging. The patients had an uneventful recovery after curettage. The follow-up time was 11 months for both patients, and no recurrence was noted. On histologic examination, the lesions were characterized by collagen-rich and generally poorly cellular tissue containing spindled to plump (myo)fibroblast-like cells and a variable admixture of inflammatory cells. Focal calcifications and reactive bone formation were present. Clonal, albeit different, chromosomal changes were found in both cases (47,XY,-9, 12,add(21)(q21),+der(?)t(?;9)(?;q11), +mar,+r and 47, XY, +r/47, idem, add(12)(p13)). The present and other reported cytogenetic findings suggest that inflammatory myofibroblastic tumors could well be neoplastic. PMID- 9331289 TI - Yolk sac tumors of the mediastinum with prominent spindle cell features: a clinicopathologic study of three cases. AB - Three cases of primary mediastinal yolk sac tumors with prominent spindle cell features are presented. The patients were three men 24-34 years of age (mean 29). Clinically, two patients presented with symptoms of chest pain and cough; no clinical information was provided for the third patient. Grossly, the tumors were described as large mediastinal masses, with a hemorrhagic and necrotic cut surface. Histologically, the tumors were characterized by a predominantly atypical spindle cell proliferation admixed with areas that showed focally the characteristic reticular growth pattern of yolk sac tumors, with the presence of Schiller-Duval bodies and intra- and extracellular hyaline globules. Immunohistochemical studies performed in one case showed positive staining for keratin and alpha-fetoprotein in both the spindle cell and reticular components of the tumor. Follow-up information was obtained in two patients; they both died of tumor with metastases to the lungs 1 year after initial diagnosis. The present cases expand the spectrum of histopathologic growth patterns that may be observed in yolk sac tumors of the mediastinum and stress the issue of careful sampling and evaluation of mediastinal neoplasms for arriving at the correct diagnosis. PMID- 9331290 TI - Vulvar Paget's disease: a clinicopathologic and immunohistochemical study of 19 cases. AB - Vulvar Paget's disease (VPD) is the most common type of extramammary Paget's disease; however, the frequency of dermal invasion and its clinical significance are unclear, as are the frequency and relationship of an associated regional internal cancer. Thus, we studied the clinicopathologic and immunohistochemical features of 19 patients with VPD. Patients ranged in age from 56 to 86 years (median 65). VPD was entirely intraepithelial (IE-VPD) in 13 patients. Three patients developed IE-VPD recurrence and one developed deeply invasive and metastatic VPD at 10.8 years. Five patients had invasive Paget's disease (INV VPD) characterized by clinically occult microscopic foci of superficial dermal invasion, ranging in depth from 0.3 to 0.9 mm. All five patients were alive without disease after 12 months to 17 years (median 66 months). A regional internal cancer (CA ASSOC-VPD) occurred in one patient whose VPD was preceded by a deeply invasive grade 3 transitional cell carcinoma of the urinary bladder 9 months earlier. Immunophenotypes of 16 cases with IE-VPD or INV-VPD were CK7+/CK20-/GCDFP15+ in 14 cases and CK7+/CK20+/GCDFP15+ in two cases, with concordant immunophenotypes of the intraepithelial and invasive components in all cases studied. The patient with CA ASSOC-VPD had a CK7+/CK20+/GCDFP15- immunophenotype in the invasive TCC of the urinary bladder and the VPD. We conclude that the predominant form of VPD begins as a primary cutaneous intraepithelial neoplasm that is universally CK7+/GCDFP15+. Foci of unsuspected synchronous dermal invasion by Paget's cells can be expected in almost one third of cases. Subsequent progression into an invasive carcinoma occurs less often. Foci of "minimally invasive" carcinoma (<1 mm) probably do not adversely affect prognosis, whereas deeply invasive carcinoma behaves as a fully malignant adenocarcinoma. The rarer form of VPD appears to result from secondary intraepithelial spread from an associated regional internal carcinoma. The finding of Paget's cells that are CK20+/GCDFP15- suggests the presence of a regional internal carcinoma with a corresponding immunophenotype. PMID- 9331291 TI - Sarcomatoid renal cell carcinoma: the chromophobe connection. AB - Eleven cases of sarcomatoid renal cell carcinoma were studied to determine the relative frequency of various subtypes of renal cell carcinoma that may be associated with sarcomatoid transformation. The epithelial components in these tumors were subcategorized according to established histologic criteria into chromophobe carcinoma (n = 6 cases), clear cell carcinoma (n = 3), papillary carcinoma (n = 1), and indeterminate (n = 1). In nine cases, material was available for immunohistochemical and DNA ploidy studies. The sarcomatoid component in all cases showed positivity for epithelial membrane antigen cytokeratin, indicating an epithelial derivation of these cells. Staining for mesenchymal markers was mostly negative, except for vimentin, which reacted strongly in all cases. DNA ploidy studies using flow cytometry and cell image analysis provided very similar results. Five of five chromophobe sarcomatoid carcinomas showed hypodiploid cell lines in the epithelial areas, whereas the sarcomatoid components mostly showed aneuploid peaks. In the remaining cases, DNA ploidy pattern was more variable. These findings indicate that chromophobe carcinoma may be the most frequent epithelial component associated with sarcomatoid renal cell carcinoma. PMID- 9331292 TI - Primary melanoma of the lung: a clinicopathologic and immunohistochemical study of eight cases. AB - Primary malignant melanoma of the lung (PMML) is an uncommon neoplasm that may be confused with more conventional types of lung cancer. Although the previously proposed criteria for diagnosis, including the presence of an in situ component, are often difficult to satisfy, this lesion is characterized by a poor prognosis, ultimately leading to patient death. We report eight cases of PMML that presented as solitary, central endobronchial neoplasms, resulting in a picture that closely resembled carcinoid tumor or poorly differentiated non-small-cell carcinoma of the lung. The mean age at diagnosis was 51 years (range 45-71). The patients included one woman and seven men. The histologic growth pattern varied from organoid to fascicular and included epithelioid to spindled cells with hyperchromatic to vesicular nuclei, prominent eosinophilic nucleoli, and abundant eosinophilic to clear cytoplasm with occasional intranuclear cytoplasmic inclusions. A bronchial in situ component was present in four cases. Initial interpretations included carcinoid tumor, non-small-cell carcinoma, and malignant melanoma. Melanin was present in all neoplasms on hematoxylin and eosin staining, although very focally in one case, and was Fontana-Masson positive in all cases. Immunohistochemically, diffuse strong positivity for S-100, HMB-45, and vimentin was present in all seven tumors tested. All seven tumors were negative for cytokeratin, CAM 5.2, and chromogranin. Ultrastructural examination of the eighth case showed dysmorphic premelanosomes but no neurosecretory granules. None of the patients had disseminated disease at presentation, and all patients underwent surgical resection (seven lobectomies and one excision). In this series, primary malignant melanoma of the lung was characterized by an aggressive postoperative course, with five patients dying of metastatic disease from 4 to 32 months after resection (median 14 months). Two patients are alive with metastatic disease at 4 and 30 months after surgery, and the eighth patient is alive with no evidence of disease 108 months after surgery at last follow-up. Metastatic melanoma was identified in various sites, including the lungs, adrenal glands, liver, mesentery, brain, and bone. The cases herein presented indicate that PMML should be included in the differential diagnosis of primary bronchial tumors. PMID- 9331293 TI - Solid variants of papillary (chromophil) renal cell carcinoma: clinicopathologic and genetic features. AB - Papillary renal cell carcinomas (RCCs) traditionally have been defined histologically as tumors with at least 50% true papillae. However, these tumors also have characteristic immunohistochemical and genetic features that separate them from other RCCs. We identified six tumors composed of solid sheets of cells without true papillae but that otherwise resembled papillary RCCs, which we feel represent solid variants of papillary RCCs. All six tumors were primary lesions of the kidney, all were strongly reactive for epithelial membrane antigen, cytokeratin 7, and callus keratin, and all were negative for the high molecular weight keratin antibody 34BE12. Four of four tumors tested showed trisomies for chromosome 7, chromosome 17, or both by either cytogenetic analysis or fluorescence in situ hybridization. Four cases were composed of solid sheets of cells containing distinct micronodules that in some cases resembled abortive papillae. The cells composing the micronodules had abundant eosinophilic cytoplasm, open chromatin, and in some cases prominent nucleoli. The intervening cells had similar nuclei, but the amount of cytoplasm was variable. In three of these micronodular cases, multiple tumors diffusely replaced the kidney; in the fourth case two typical clear cell RCCs and a typical papillary RCC were also present in the same kidney, but the micronodular tumor was unifocal. The two remaining cases were solitary tumors consisting of solid sheets of cells forming ill-defined tubules. These cells had scant clear cytoplasm and small round to elongate nuclei with occasional nuclear grooves but only rare small nucleoli. Limited follow-up has shown no evidence of disease in any patient thus far. The differential diagnosis includes "renal adenoma," renal adenomatosis, metanephric adenoma, and clear/granular cell RCC. We conclude that solid variants of papillary RCCs lack true papillae but have characteristic histologic, immunohistochemical, and genetic features. PMID- 9331294 TI - Hepatoid yolk sac tumors of the mediastinum: a clinicopathologic and immunohistochemical study of four cases. AB - Four cases of primary hepatoid yolk sac tumors of the anterior mediastinum are described. The patients were all men between the ages of 26 and 40 years (median 33). Clinically, they all presented with a history of shortness of breath and chest pain of several weeks' duration. None of the patients had a history of germ cell tumor elsewhere or evidence of any hepatic abnormality. Grossly, all the tumors were described as large mediastinal masses that impinged on adjacent structures. Histologically, they were characterized by sheets of medium-sized, round to polygonal neoplastic cells with moderate amounts of eosinophilic cytoplasm and round to oval nuclei with prominent nucleoli. The cellular proliferation was homogeneous and displayed moderate cellular atypia and scattered mitotic activity. All the tumors showed focally the presence of more conventional areas of yolk sac tumor, with islands of tumor cells showing a reticular pattern of growth admixed with scattered intra- and extracellular hyaline globules and occasional Schiller-Duval bodies. Immunohistochemical studies showed strong positivity of the tumor cells for alpha-fetoprotein in both components of the lesions. Follow-up information was available in three patients, all of whom developed lung metastases within a year after initial diagnosis. Two of these patients died of tumor within the same period, whereas a third patient has been lost to follow-up. The present cases illustrate an unusual histologic pattern of yolk sac tumor in the mediastinum and highlight the importance of considering this tumor in the differential diagnosis of lesions showing a hepatoid pattern of growth in the mediastinal area. PMID- 9331295 TI - Unusual histologic types of high-grade prostatic intraepithelial neoplasia. AB - High-grade prostatic intraepithelial neoplasia (HGPIN) is the most likely precursor proliferation of peripheral zone, moderately to poorly differentiated prostatic adenocarcinomas. The usual cell type of the epithelial lining of HGPIN is a glandular epithelial cell with characteristic nuclear abnormalities. Here we report nine cases of unusual types of HGPIN, including three cases of signet-ring cell HGPIN, one case of small cell neuroendocrine HGPIN, and five cases of HGPIN with distinctive mucinous features. The three examples of signet-ring cell PIN were all associated with an invasive primary signet-ring cell carcinoma of the prostate. The HGPIN assumed a classical tufted and micropapillary architectural growth pattern, with the constituent cells exhibiting a morphologic appearance identical to that of the invasive signet-ring cells. The intraepithelial and invasive signet-ring cells were mucin negative and were immunoreactive for prostate-specific antigen (PSA). A fourth case displayed a mixed intraepithelial glandular-small cell neoplastic proliferation, where intraepithelial small cells were histologically identical to surrounding invasive small cell carcinoma cells. The small cell HGPIN and invasive small cell carcinoma cells were positive for the neuroendocrine markers chromogranin, synaptophysin, and neuron-specific enolase. In five cases, mucinous distension of HGPIN glands, producing a flat pattern of the epithelial lining layer, comprised the third unusual pattern of HGPIN. These blue mucinous secretions were readily detected by hematoxylin and eosin staining and were composed of both neutral (periodic acid-Schiff-positive) and acidic (alcian blue-positive) mucins. Herein we document the existence of an intraepithelial proliferation of neoplastic cell types-small cell neuroendocrine and signet-ring cell-that are usually considered as stromal-invasive cells in the prostate. The presence of these rare prostatic cell types in both HGPIN and invasive carcinoma provides further support for a close relationship between HGPIN and invasive carcinoma of the prostate. All three unusual types of HGPIN signet-ring cell, small cell neuroendocrine, and mucinous-are important to diagnostically recognize because of the strength of association of HGPIN with invasive carcinoma. PMID- 9331296 TI - Blastic natural killer cell leukemia/lymphoma: a clinicopathologic study. AB - The classification of natural killer (NK)-cell and NK-like T-cell malignancies has undergone significant evolution in recent years. Although examples of NK-cell tumors resembling acute leukemia have been described anecdotally as blastic, blastoid, or monomorphic NK-cell leukemia/lymphoma (NKL/L), the clinical and pathologic features of these tumors have not been systematically defined. We report four patients with blastic NKL/L and describe the clinical, pathologic, and immunophenotypic findings in these cases. All patients were elderly (58-82 years) and presented with cutaneous plaques. Two patients also had adenopathy, and three patients had marrow involvement at presentation. Biopsy of cutaneous lesions showed atypical superficial and deep dermal lymphoid infiltrates. Involved lymph nodes were architecturally effaced by an interfollicular infiltrate with blastic cytologic features. In Wright-Giemsa-stained blood or marrow smears, tumor cells had finely distributed nuclear chromatin, many with nucleoli, and variable amounts of cytoplasm. In contrast to many NK and NK-like T cell disorders, azurophilic cytoplasmic granules were absent or inconspicuous. The tumor cells were immunophenotypically distinctive. They expressed intermediate density CD45, as is characteristic of blasts; in addition, the cells were positive for HLA-DR, CD2, CD4, and the NK-associated antigen CD56. Surface CD3, cytoplasmic CD3, and CD5 were negative in all cases tested, whereas CD7 was expressed in two cases. In formalin-fixed tissue, tumor cells marked with antibodies to CD43, but not with other T- or B-lineage-related antibodies. All three cases studied for Epstein-Barr viral RNA by in situ hybridization were negative. Although treatments varied, all three patients with clinical follow-up died within months of the diagnosis. The clinical course in two patients culminated in an overtly leukemic phase. These findings suggest that blastic NKL/L represents a distinct clinicopathologic entity, characterized by cutaneous, nodal, and marrow involvement by blastic cells with immunophenotypic characteristics of true NK cells. The disease afflicts elderly patients, pursues an aggressive course, and may culminate in overt leukemia. PMID- 9331297 TI - Infantile hemangioendothelioma of the ovary: a monodermal teratoma or a neoplasm of ovarian somatic cells? AB - Vascular tumors of the female genital tract are uncommon, and only a few cases have been reported in the ovary. We describe herein, an unusual tumor of the ovary: infantile hemangioendothelioma (cellular hemangioma of infancy) in a newborn. The tumor consisted of well-formed blood vessels and proliferating endothelial cells that were arranged in solid cordlike structures. The tumor permeated the ovarian stroma and entrapped normal ovarian follicles. By immunohistochemistry the neoplastic cells expressed factor VIII, CD34, and alpha smooth-muscle actin, and ultrastructurally they had the features of endothelial cells that were focally associated with pericytes. We examined simple sequence repeat (SSR) polymorphic markers in the tumor tissue, as well as in the patient's and parents' blood. The informative SSR markers were found to be identical in the tumor and in the patient's somatic cells. We suggest that the tumor described herein is a congenital infantile hemangioendothelioma arising from ovarian parenchymal cells rather than a teratoma originating from germ cells. A similar morphologic lesion has been described recently in the ovary and interpreted as monodermal teratoma composed of vascular tissue. PMID- 9331299 TI - Clear cell "sugar" tumor of the lung: association with lymphangioleiomyomatosis and multifocal micronodular pneumocyte hyperplasia in a patient with tuberous sclerosis. AB - We report the unique association of a clear cell "sugar" tumor of the lung (CCTL) in a 32-year-old woman with tuberous sclerosis (TSC), lymphangioleiomyomatosis (LAM), and multifocal micronodular pneumocyte hyperplasia (MMPH). Chest radiographs demonstrated a peripheral solitary 1.0-cm lingular nodule, diffuse emphysematous changes, and bilateral pneumothorax. Microscopic examination of the coin lesion showed an unencapsulated tumor composed of round to oval to focally spindled cells with distinct cellular borders, abundant clear to eosinophilic granular cytoplasm, prominent thin-walled blood channels, and focal hyaline stroma. Rare multinucleated cells were identified, and neither necrosis nor mitotic figures were seen. Tumor cells contained abundant diastase-sensitive intracytoplasmic glycogen, as demonstrated with periodic acid-Schiff stains. Tumor cell immunoreactivity for HMB-45 and nonreactivity for cytokeratin supported the diagnosis. The lung tissue also contained MMPH and smooth muscle proliferations diagnostic of LAM. The histogenesis of CCTL remains controversial, and similarities between this lesion and both LAM and angiomyolipoma (AML) raise the possibility that these lesions are related not only to each other, but that CCTL should be added to the spectrum of pulmonary manifestations of TSC. PMID- 9331298 TI - Anaplastic large cell lymphoma of maternal origin involving the placenta: case report and literature survey. AB - Non-Hodgkin's lymphoma (NHL) occasionally involves the placenta, and information of such occurrence should be useful for management of the mother and fetus. We report the first case of anaplastic large cell lymphoma (ALCL) disseminated to the placenta. The diagnosis was made via excisional biopsy of cervical lymphadenopathy in a 20-year-old woman at 27 weeks' gestation. Involvement of the placenta was noted on gross examination after cesarean section delivery of a girl at 30 weeks' gestation. The ALCL was microscopically confined to intervillous spaces in a manner similar to previous reports of other NHLs. The immunophenotype was characteristic (CD30+, EMA+, BNH9+), and the now frequently associated t(2;5)(p23;q35) translocation with this lymphoma was detected by the recently produced monoclonal antibody ALK1 against the nucleophosmin/anaplastic lymphoma kinase (NPM/ALK) chimeric protein. Complete remission was induced in the mother after delivery. Both mother and child are healthy at 10 years' follow-up. The case is reported in light of the sparse literature on lymphomatous involvement of the placenta. PMID- 9331300 TI - Hepatic undifferentiated (embryonal) sarcoma arising in a mesenchymal hamartoma. AB - We report the case of a hepatic undifferentiated (embryonal) sarcoma (UES) arising within a mesenchymal hamartoma (MH) in a 15-year-old girl. Mapping of the tumor demonstrated a typical MH transforming gradually into a UES composed of anaplastic stromal cells. When evaluated by flow cytometry, the MH was diploid and the UES showed a prominent aneuploid peak. Karyotypic analysis of the UES showed structural alterations of chromosome 19, which have been implicated as a potential genetic marker of MH. The histogenesis of MH and UES is still debated, and reports of a relationship between them, although suggested on the basis of histomorphologic similarities, have never been convincing. The histologic, flow cytometric, and cytogenetic evidence reported herein suggests a link between these two hepatic tumors of the pediatric population. PMID- 9331301 TI - Papillary thyroid carcinoma with lipomatous stroma. PMID- 9331302 TI - Is cotyledonoid dissecting leiomyoma of the uterus (Sternberg tumor) identical with grapelike leiomyoma of the uterus? PMID- 9331303 TI - Is cotyledonoid dissecting leiomyoma of the uterus (Sternberg tumor) identical with grapelike leiomyoma of the uterus? PMID- 9331304 TI - The Triological Society: cradle of certification. PMID- 9331305 TI - A rationale for therapy of the N0 neck. PMID- 9331306 TI - Survival, function, and quality of life after total glossectomy. AB - Advanced tongue cancer is associated with poor survival despite aggressive therapy. In an attempt at cure, many patients undergo total glossectomy, which significantly affects function and quality of life (QOL). This study was designed to determine the survival rate and QOL of patients who had undergone total glossectomy. A total of 54 patients underwent total glossectomy, with or without total laryngectomy, for advanced tongue cancer from 1970 to 1996. Patient outcomes were assessed for the following: 1. disease-free survival, 2. function, utilizing the Performance Status Scale (PSS), and 3. QOL, using two general cancer questionnaires (FACT-G and EORTC QLQ-C30) and a series of questions specific for head and neck cancer patients. Corrected actuarial survival was 51% and 41% at 3 and 5 years, respectively. Functional assessment using the PSS demonstrated significant deficits in speech and deglutition. QOL questionnaires revealed problems with eating, speaking, socializing, and shoulder function. However, the overall responses demonstrated that these patients have adjusted to their deficits and have a good QOL. It was concluded that total glossectomy, with or without total laryngectomy, can result in meaningful survival and an adequate QOL can be achieved in selected patients. PMID- 9331307 TI - Management of nasopharyngeal and oropharyngeal stenosis in children. AB - Nasopharyngeal stenosis and oropharyngeal stenosis are rare and challenging problems in the pediatric population. The most common etiology is currently the surgical trauma associated with adenotonsillectomy. Stenosis can vary from a thin band to a complete obstructing cicatrix. Presenting symptoms range from mild hyponasal speech to severe airway obstruction. We present a series of eight children with varying degrees of stenosis and associated symptoms. Choice of treatment varied with the severity of disease. In our series, successful interventions included triamcinolone acetonide injection, lysis of adhesions, rotational and advancement mucosal flaps, and jejunal free flap. Preoperative evaluation and individualized surgical repair are essential for successful treatment. PMID- 9331308 TI - An optimal choice: home intravenous hydration after tonsillectomy. AB - Most tonsillectomies are performed as outpatient procedures in an ambulatory care facility. The postoperative period can be protracted, which can cause dehydration and readmission to the hospital. Timely intervention with home intravenous (IV) hydration can help prevent this complication. A prospective clinical trial was designed to evaluate the benefit of home IV therapy after tonsillectomy. One hundred consecutive patients underwent tonsillectomy. Fifty returned home without home IV hydration and another 50 returned home with home IV hydration. Clinical data were collected by the homecare nurse during the postoperative period. Comparison between these two groups showed that the patients with home IV hydration had less morbidity. Daily assessment of the patient by telephone is the best screening method to determine the need for IV hydration for the posttonsillectomy patient. PMID- 9331309 TI - Fibrous dysplasia of the temporal bone: reversal of sensorineural hearing loss after decompression of the internal auditory canal. AB - When fibrous dysplasia affects the temporal bone, it most often presents with conductive hearing loss attributable to stenosis of the external auditory canal. Sensorineural hearing loss has usually been attributed to involvement of the otic capsule. We present a patient with bilateral fibrous dysplasia of the temporal bones who complained of unilateral hearing loss, facial tingling, and facial twitching. The audiogram showed severe sensorineural hearing loss. The hearing markedly improved and facial twitching and tingling ceased after decompression of the internal auditory canal via a middle fossa approach. This is the only case of which we are aware showing reversal of sensorineural hearing loss caused by fibrous dysplasia. PMID- 9331310 TI - Endoscope-assisted second-stage tympanomastoidectomy. AB - Intact canal wall mastoidectomy techniques for cholesteatoma are often followed by a planned second look for residual disease and possible ossicular reconstruction. Endoscopic techniques may reduce morbidity but introduce new concerns. Twenty-five consecutive second-look procedures were performed from July 1994 to July 1996 utilizing endoscopes in 19 cases and avoiding or terminating their use in the others because of known difficult anatomy, inadequate exposure, or excessive bleeding. Thirteen cases were prospectively explored first through a planned exclusively endoscopic approach and then opened for a conventional second look in comparison. In one of the 13 cases, endoscopy was abandoned. There were no cases in which endoscopy yielded a false-negative result. Endoscopes underestimated the size of recurrence in one case. Our experience, indications, and precautions for endoscope-assisted second-stage tympanomastoidectomy are presented. PMID- 9331311 TI - Validation of objective measures for facial paralysis. AB - A shift from subjective scales to objective measures of facial paralysis requires physical models against which to validate and calibrate the new objective techniques. The purpose of this report was to demonstrate such a model and to test the capacity of an objective computer system to accurately measure physical change. The physical model consisted of an advancing edge of a spreading gelatin film. The model was measured in submillimeter increments. Concurrent measurements were made using the computed system. Ten trials were conducted. The objective system proved to be exquisitely sensitive (0.03 mm) and highly correlated with the physical criterion model (Pearson's product moment correlation coefficient [r]=0.9849). Sensitive and calibrated objective methods of analysis of facial motion are crucial to the design of clinical trials, clinical studies of nerve regeneration, and comparisons of reanimation techniques. PMID- 9331312 TI - Inner and middle ear hyperbaric oxygen-induced barotrauma. AB - Patients receiving hyperbaric oxygen (HBO) therapy can sustain inner and middle ear barotrauma. The purpose of this study is to define the incidence and significance of HBO-related barotrauma, in addition to establishing guidelines for prophylactic myringotomy or tympanostomy tube placement. Thirty patients were stratified into two groups (those able to autoinflate and those unable to autoinflate the middle ear) and barotrauma was assessed by otoscopy, tympanometry, high-frequency audiometry, and distortion product otoacoustic emission (DPOAE) testing. Ten of 11 patients (91%) from the noninflater group suffered middle ear barotrauma, and seven of 19 patients (37%) from the autoinflater group sustained middle ear barotrauma. Patients unable to autoinflate the middle ear were shown to have a higher incidence and greater severity of barotrauma than patients able to autoinflate. Pretreatment pressure equalizing tubes or myringotomies should be considered for patients undergoing HBO therapy who have an artificial airway or have eustachian tube dysfunction and have failed conservative medical intervention. A significant change in DPOAEs (loss of emissions over a 1-kHz range) was found in four of 15 autoinflaters (27%) and two of seven noninflaters (29%). There was no significant difference between the groups. The decrease in DPOAEs was not associated with a change in conventional audiometry. PMID- 9331313 TI - Interinterpreter variability in determining the SP/AP ratio in clinical electrocochleography. AB - Calculating the SP/AP ratio in electrocochleography requires the interpreting audiologist to unequivocally identify the peak of the SP wave form. If different points are selected, different SP/AP ratios will result. To investigate this effect, 50 electrocochleographic tracings were sent to 10 different audiologists (500 tracings). Twenty of the wave forms sent to each audiologist were identified as "easy," 10 as "somewhat difficult," 10 as "very difficult," and 10 as "no response." The ranges and standard deviations of the resulting SP/AP ratios were quite high. Analysis of variance showed statistically significant differences in the very difficult and no response group. There is significant interinterpreter difference among SP/AP ratios calculated from the same tracing. This variability affects clinical patient management and investigational projects. PMID- 9331314 TI - Translacrimal transnasal laser-assisted dacryocystorhinostomy. AB - Chronic dacryocystitis is due to an obstruction in the nasolacrimal duct, with subsequent infection of the lacrimal sac. The goal of surgery is to reestablish intranasal drainage of the lacrimal sac. Classic dacryocystorhinostomy (DCR) requires an external incision and drilling through the lacrimal bone into the middle meatus. In our study a 600-micron neodymium:YAG (Nd:YAG) fiber with a blunt hemispherical tip is inserted via the lacrimal puncta. An intranasal ostium is created with the laser under intranasal endoscopic control. Silicon tubes are then left in place for 6 months. We have performed 49 procedures over the past 2 1/2 years, with a success rate of 85% after one surgical procedure, which is commensurate with standard DCR. This procedure provides a simple, bloodless, incisionless alternative to standard DCR. PMID- 9331315 TI - Ansa cervicalis nerve: review of the topographic anatomy and morphology. AB - In recent years, there has been a proliferation of techniques utilizing the ansa cervicalis nerve to reinnervate the paralyzed larynx. The anatomic course and morphology of the ansa cervicalis are complicated by the variable course and location along the great vessels of the neck, as well as the significant differences observed in the arrangement of its contributing roots and regional branching patterns. Herein, we review the surgical anatomic course of ansa cervicalis and its innervation of the muscles of the neck, and develop specific recommendations with respect to the use of this nerve in laryngeal reinnervation. PMID- 9331316 TI - Gastroesophageal reflux, motility disorders, and psychological profiles in the etiology of globus pharyngis. AB - The aim of this study was to investigate the origin of globus pharyngis with particular reference to esophageal disorders such as gastroesophageal reflux disease (GERD), motility disorders, structural abnormalities, other gastrointestinal tract diseases, and psychological profile. Previous studies on this subject using 24-hour pH monitoring give conflicting results and are hampered by the high background prevalence of asymptomatic GERD in the normal Western population. The local Chinese population is known to have a very low background level of GERD and therefore is an ideal study population. Twenty-six patients with globus pharyngis underwent 24-hour ambulatory pH monitoring, esophageal manometry, and esophagogastroduodenoscopy with lower esophageal biopsy. A control group of 20 patients presenting with non-ulcer dyspepsia was similarly investigated. Personality profiles of the globus pharyngis subjects and an appropriate control group were assessed. Eight of the globus pharyngis group (30.7%) had evidence of GERD, whereas only one of the controls (5%) demonstrated GERD on 24-hour esophageal pH monitoring (P < 0.05). The manometric and personality profile studies did not show significant differences between study and control groups. We concluded that the finding of GERD in patients with globus pharyngis is not a coincidental finding but that there is a true association between GERD and globus pharyngis. PMID- 9331317 TI - Sustained release of antibiotic from a fibrin-gelatin-antibiotic mixture. AB - To improve the therapeutic effect of antibiotics in the ear cavity postoperatively, a fibrin- and gelatin-based drug delivery system was designed and in vitro efficacy was evaluated. Four kinds of fibrin clot were made from ampicillin, gentamicin, and ofloxacin, and biologic assay was performed using Bacillus subtilis. The fibrin-gelatin-gentamicin mixture demonstrated antibiotic activity for up to 120 hours against B subtilis. Gentamicin was found to be released more slowly from the fibrin-gelatin mixture than from the fibrin mixture in this experiment. These results suggest that this device can be clinically used in a limited field such as postoperative care of ear infection. PMID- 9331318 TI - Do systemic corticosteroids effectively treat obstructive sleep apnea secondary to adenotonsillar hypertrophy? AB - To determine if pediatric obstructive sleep apnea syndrome (OSAS) caused by adenotonsillar hypertrophy (ATH) could be treated by a short course of systemic corticosteroids, we conducted an open-label pilot study in which standardized assessments of symptomatology, OSAS severity, and adenotonsillar size were performed before and after a 5-day course of oral prednisone, 1.1+/-0.1 (+/-SE) mg/kg per day. Outcome measures included symptom severity, adenotonsillar size, and polysomnographic measures of OSAS. Selection criteria included age from 1 to 12 years, ATH, symptomatology suggesting OSAS, an apnea/hypopnea index (AHI) > or = 3/hour, and intent to perform adenotonsillectomy. Only one of nine children showed enough improvement to avoid adenotonsillectomy. Symptomatology did not improve after corticosteroid treatment but did after removal of tonsils and adenoids. Polysomnographic indices of OSAS severity did not improve after corticosteroid treatment. After corticosteroids, tonsillar size decreased in only two patients, adenoidal size was only marginally reduced, and the size of the nasopharyngeal airway was not significantly increased. These results suggest that a short course of prednisone is ineffective in treating pediatric OSAS caused by ATH. PMID- 9331319 TI - Rethinking the use of auditory brainstem response in acoustic neuroma screening. AB - The ability of magnetic resonance imaging (MRI) to detect very small acoustic tumors has triggered many to rethink the use of auditory brainstem response (ABR) in the screening of acoustic tumors. To assess ABR accuracy, we conducted a retrospective study of 388 surgically treated patients. Of these patients, 111 had complete databases including both preoperative MRIs and ABRs. The ABR was abnormal by wave V interaural latency difference in 106 (95%) of the cases. Although our overall sensitivity was 95%, sensitivity varied according to tumor size. ABR was abnormal or absent for all tumors (100%) larger than 2 cm in diameter, for 98% of tumors 1.1 to 2 cm in diameter, and for only 89% of tumors less than or equal to 1 cm in diameter. Ramifications of this in the decision making process are presented. Criteria for cut-off values are also discussed. PMID- 9331320 TI - Plasticity of the extracranial facial nerve. AB - Although the expansion properties of peripheral nerves have been a matter of considerable study in recent years, investigations of the plasticity of cranial nerves, including the facial nerve, have been lacking. Clinicians, however, have long recognized the tenacity of facial nerve function in patients with slow growing benign tumors that enormously distort the nerve. An experimental study was designed to assess whether tissue expansion techniques can be applied to the extracranial portion of the facial nerves of cats. In eight cats the frontozygomatic branch of the facial nerve was expanded by stages in seven sessions over a period of 40 days. The length of the nerve increased an average of 95% without significantly impairing nerve function. Pressure changes in the expander averaged 75 mm Hg during each stage of expansion. Electroneurography was performed after each injection of the expander. Statistical analysis of these data did not show consistent evidence of demyelination or denervation, and all but one cat exhibited a normal blink reflex and had normal electromyographic findings at the end of the experiment. Histologic examination of the expanded nerves, however, did show inflammatory changes, intraneural edema, and occasional demyelination. PMID- 9331321 TI - Murine model of interleukin-8-induced otitis media. AB - Interleukin-8 (IL-8), a potent inflammatory mediator that is thought to control leukocyte recruitment and activation during inflammatory reactions, has been implicated in a variety of inflammatory diseases. Recent studies in our laboratory have demonstrated the presence of IL-8 in chronically inflamed human middle ear effusions. These data have led us to hypothesize that IL-8 is responsible for the leukocyte recruitment seen in otitis media. Because the effect of IL-8 on the middle ear mucosa has not been investigated and therefore its role in middle ear inflammation is not known, we investigated the ability of IL-8 to directly induce inflammation in the murine middle ear. For these studies, ICR mice were used to test the hypothesis that IL-8 could directly induce inflammation in the middle ear. Murine middle ears received 8-mL transtympanic injections of human IL-8 (25 mg/mL) in saline, heat-killed Streptococcus pneumoniae (1 x 10(8)/mL), or normal saline. Temporal bones were removed at 1, 4, 8, 24, and 48 hours, decalcified, and processed for histologic examination. Noninjected murine temporal bones served as controls. Normal saline-injected ears demonstrated little to no change as compared with temporal bones from noninjected ears. IL-8-injected ears histologically demonstrated thickening of the epithelial layer and subepithelial space (SES), with inflammatory cell infiltration in the SES peaking at 4 to 8 hours and resolving by 48 hours. Bacteria-injected ears demonstrated findings similar to, although not as extensive as, those found in IL 8-injected ears (i.e., inflammatory reactions peaked at 8 to 24 hours and resolved by 72 hours). Our results demonstrate that IL-8 is a potent inducer of middle ear inflammation and support the concept that IL-8 may be one of the key cytokines responsible for the leukocyte accumulation and activation seen in otitis media. PMID- 9331322 TI - Effects of systemic hyperoxia on eustachian tube ventilatory function. AB - Middle ear negative pressure and effusions, decreased middle ear compliance, and abnormal tympanometry results have been described after diving on oxygen. Middle ear gas hyperoxia has been shown to down-regulate the eustachian tube ventilatory function (ETVF). The purpose of the present study was to investigate to what extent systemic hyperoxia in the face of air-equivalent middle ear gas composition might interfere with the ETVF. ETVF was investigated in four young adult female cynomolgus monkeys by the forced-response and inflation-deflation tests using air while the animals breathed either room air or 100% normobaric oxygen. Higher opening, closing, and steady-state pressures were observed under systemic hyperoxia. The percentage of the applied pressure equalized, and the maximal pressure change on a single swallow in the deflation test were both lower under hyperoxic conditions. The results show that systemic hyperoxia might impair ETVF. This observation adds to our understanding of the pathophysiology of middle ear dysfunction observed after diving on oxygen. PMID- 9331323 TI - Monitoring eustachian tube opening: preliminary results in normal subjects. AB - Sonometry studies the changes of sound intensities induced by the eustachian tube opening in the external meatus when a constant sound is applied in the nasal cavity. The function of the eustachian tube is not disturbed by the test procedure. An original portable device with a high signal-to-noise ratio has been developed in our lab. This device was able to detect a 6-dB sound pressure level (SPL) signal in a 100-dB SPL noise. Long-duration recordings of auditory tubal openings could be performed. This work presents the data found in normal subjects. One hundred twenty healthy ears were recorded. Eustachian tube openings were detected in 62.5% of the cases. Of the 1777 openings recorded, the average opening duration was 430 ms +/- 183 ms. The average number of eustachian tube openings was 21.7+/-16.4 (for 15 minutes). All swallowing did not necessary open the eustachian tube. This work shows that this new device allows long-duration recording of eustachian tube function in everyday life conditions. PMID- 9331324 TI - Sphenopalatine blocks in the treatment of pain in fibromyalgia and myofascial pain syndrome. AB - Sphenopalatine blocks have been used to treat pain for more than 80 years. Anecdotal support for sphenopalatine ganglion blocks has been very strong in those who believe in the technique, but the research results have been inconclusive. Therefore, a double blind, placebo-controlled study was performed on 61 patients, 42 with fibromyalgia and 19 with myofascial pain syndrome. Pain was measured using visual analogue scales prior to treatment, during treatment, and 28 days after the treatment. Headaches were evaluated in frequency and location prior to and after treatment. Sphenopalatine ganglion blocks were performed under direct vision using 4% lidocaine and sterile water as a placebo. Analysis of the results showed no statistical differences between the lidocaine and the placebo groups. PMID- 9331325 TI - Extended middle meatal antrostomy. PMID- 9331327 TI - xnf7 functions in dorsal-ventral patterning of the Xenopus embryo. AB - Xenopus nuclear factor 7 (xnf7) is a maternally expressed nuclear protein that is retained in the cytoplasm from oocyte maturation until the midblastula transition (MBT). Mutations of the xnf7 phosphorylation sites to glutamic acids (dnxnf7) resulted in the retention of the endogenous protein in the cytoplasm past the MBT, indicating that cytoplasmic retention is a phosphorylation dependent process. In addition, dnxnf7 acted as a dominant negative mutant by keeping the endogenous xnf7 protein in the cytoplasm past the MBT. Overexpression of dnxnf7 in future dorsal blastomeres resulted in a ventralized or posteriorized phenotype in which the embryos lacked anterior structures, while overexpression in ventral blastomeres resulted in dorsalized embryos. dnxnf7 also affected the expression of both dorsal and ventral mesodermal markers. These data suggest that xnf7 functions in dorsal/ventral patterning and that the movement of the protein from the cytoplasm to the nucleus at the MBT is critical for the execution of a genetic program conferring a dorsal or ventral identity to the mesoderm. PMID- 9331329 TI - Interspecies exchange of a Hoxd enhancer in vivo induces premature transcription and anterior shift of the sacrum. AB - The precise activation, in space and time, of vertebrate Hox genes is an essential requirement for normal morphogenesis. In order to assess for the functional potential of evolutionary conserved Hox regulatory sequences, a phylogenetically conserved bipartite regulatory element necessary for proper spatial and temporal activation of the Hoxd-11 gene was replaced by its fish counterpart in the HoxD complex of mice, using an ES cell-based targeted exchange. Fetuses carrying this replacement activated Hoxd-11 transcription prematurely, which led to a rostral shift of its expression boundary and a consequent anterior transposition of the sacrum. These results demonstrate the high phylogenetic conservation of regulatory mechanisms acting over vertebrate Hox complexes and suggest that minor time difference (heterochronies) in Hox gene activation may have contributed to important morphological variations in the course of evolution. PMID- 9331328 TI - Inhibitory effects of ventral signals on the development of Brn-3.0-expressing neurons in the dorsal spinal cord. AB - Brn-3.0, a POU-domain transcription factor, is expressed in specific postmitotic neurons in the dorsal part of the neural tube which are among the first spinal cord neurons to appear in development. In the mature spinal cord, the Brn-3.0 cells form a numerous population of scattered neurons in the intermediate spinal gray. Ablation of the notochord in chick embryos extends the domain of Brn-3.0 expression into the ventral neural tube, while ectopic grafts of notochord tissue suppress Brn-3.0 expression. The notochord effects on Brn-3.0 expression are reproduced in vivo by the implantation of a local source of recombinant Shh protein. The down-regulation of Brn-3.0 expression in the dorsal spinal cord by the notochord and Shh contrasts with the known inductive effects of these ventral signals on the approximately simultaneous development of the spinal motor neurons. In cultured explants of neural plate from the region of the presumptive spinal cord, Brn-3.0 neurons develop in the absence of surface ectoderm and ventral midline tissue, suggesting that the Brn-3.0 phenotype may represent a "default" developmental pathway for early spinal cord neurons. Together these results advance the understanding of the mechanism of the generation of neuronal diversity in the developing vertebrate CNS. PMID- 9331330 TI - Intracellular pH increase driven by an Na+/H+ exchanger upon activation of surf clam oocytes. AB - Intracellular pH (pHi) measurements were performed in surf clam (Spisula solidissima) oocytes before and after artificial activation or fertilization [evidenced by germinal vesicle breakdown (GVBD)] by the dimethyloxazolidinedione (DMO) and 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) methods. Results using both methods showed increases of pHi of 0.3 pH unit after activation by excess K+. Using BCECF, we found an increase of similar magnitude after fertilization or after the addition of serotonin. By contrast, GVBD did not occur when the pHi was increased to similar or even higher levels by exposing the oocytes to ammonia. In sodium-free seawater, excess K+ induced GVBD but the pHi of K+-activated oocytes decreased significantly below the resting level of unactivated oocytes. The pHi increases in K+-activated oocytes were otherwise proportional to the external Na+ concentration. The amiloride derivatives dimethylamiloride and hexamethylene amiloride (at 10-50 microM) efficiently inhibited the K+-induced increase of pHi but did not block GVBD. These two derivatives were able, however, to retard K+-induced GVBD, hexamethylene amiloride being the more efficient. This retardation of K+-induced GVBD could be abolished by the simultaneous addition of ammonia. Taken altogether, these results show that a pHi increase, driven by a typical Na+/H+ exchanger, follows activation of surf clam oocytes but that this pHi increase is neither sufficient nor required for GVBD, though it does allow its progression at an optimal rate. PMID- 9331331 TI - Mash2 acts cell autonomously in mouse spongiotrophoblast development. AB - The Mash2 gene, which encodes a basic helix-loop-helix transcription factor, is one of the mammalian homologues of the Drosophila achaete-scute genes. It is strongly expressed in diploid trophoblast cells of the postimplantation mouse embryo. Targeted mutagenesis of Mash2 revealed that loss of function results in embryonic lethality at midgestation, due to placental failure associated with a lack of spongiotrophoblast and reduced labyrinthine trophoblast layers. For the further study of Mash2 function in development of the trophoblast cell lineage, we have performed chimeric analysis combining Mash2 mutant and wild-type embryos. We have addressed the question of whether the phenotype of the Mash2 mutant embryo, which affects all of the three trophoblast cell layers, is caused by a cell autonomous or non-autonomous defect and whether Mash2 is required in both spongiotrophoblast and labyrinthine trophoblast development. Our results showed no contribution of Mash2 mutant cells to the spongiotrophoblast layer in chimeric placentae at 10.5 and 12.5 days postcoitum, suggesting that the product of the Mash2 gene is required cell autonomously during the development of the spongiotrophoblast. However, it seems that Mash2 is not required for development of labyrinthine trophoblast or giant cells, since high contributions of Mash2 mutant cells were observed in those trophoblast cell layers in the chimeric placentae analyzed. We can therefore conclude that the primary and cell autonomous function of Mash2 appears to be an involvement in the development of diploid trophoblast cells in the ectoplacental cone to form the spongiotrophoblast cell layer of the mature chorioallantoic placenta. PMID- 9331332 TI - Genetic and functional analysis of neuronatin in mice with maternal or paternal duplication of distal Chr 2. AB - Functional differences between parental genomes are due to differential expression of parental alleles of imprinted genes. Neuronatin (Nnat) is a recently identified paternally expressed imprinted gene that is initially expressed in the rhombomeres and pituitary gland and later more widely in the central and peripheral nervous system mainly in postmitotic and differentiating neuroepithelial cells. Nnat maps to distal chromosome (Chr) 2, which contains an imprinting region that causes morphological abnormalities and early neonatal lethality. More detailed mapping analysis of Nnat showed that it is located between the T26H and T2Wa translocation breakpoints which is, surprisingly, proximal to the reported imprinting region between the T2Wa and T28H translocation breakpoints, suggesting that there may be two distinct imprinting regions on distal chromosome 2. To investigate the potential role of Nnat, we compared normal embryos with those which were PatDp.dist2.T26H (paternal duplication/maternal deficiency of chromosome 2 distal to the translocation breakpoint T26H) and MatDp.dist2.T26H. Expression of Nnat was detected in the PatDp.dist2.T26H embryos, where both copies of Nnat are paternally inherited, and normal embryos but no expression was detected in the MatDp.dist2.T26H embryos with the two maternally inherited copies. The differential expression of Nnat was supported by DNA methylation analysis with the paternally inherited alleles being unmethylated and the maternal alleles fully methylated. Although experimental embryos appeared grossly similar phenotypically in the structures where expression of Nnat was detected, differences in folding of the cerebellum were observed in neonates, and other more subtle developmental or behavioral effects due to gain or loss of Nnat cannot be ruled out. PMID- 9331333 TI - Expression of the cell surface proteoglycan glypican-5 is developmentally regulated in kidney, limb, and brain. AB - Heparan sulfate is ubiquitous at the cell surface, where it is expressed predominantly on proteoglycans of either the transmembrane syndecan family or the glycosylphosphatidylinositol (GPI)-anchored glypican family, and has been proposed to function as a "coreceptor" for a number of "heparin-binding" growth factors. Although little is known about functional differences between individual members of the glypican gene family, mutations in both the Drosophila gene dally and the human gene for glypican-3 strongly suggest that at least some glypicans do function in cellular growth control and morphogenesis. In particular, deletion of the human glypican-3 gene is responsible for Simpson-Golabi-Behmel syndrome, and its associated pre- and postnatal tissue overgrowth, increased risk of embryonal tumors during early childhood, and numerous visceral and skeletal anomalies. We have identified and characterized, by sequencing of EST clones and products of rapid amplification of cDNA ends (RACE), an mRNA that encodes a 572 amino-acid member of the glypican gene family (glypican-5) that is most related (50% amino acid similarity, 39% identity) to glypican-3. Glypican-5 mRNA is detected as a 3.9- and 4.4-kb transcript in adult and neonatal mouse brain total RNA, and in situ hybridization results localize transcript primarily to restricted regions of the developing central nervous system, limb, and kidney in patterns consistent with a role in the control of cell growth or differentiation. Interestingly, glypican-5 localizes to 13q31-32 of the human genome, deletions of which are associated with human 13q- syndrome, a developmental disorder with a pattern of defects that shows significant overlap with the pattern of glypican-5 expression. PMID- 9331335 TI - Characterization of the sea urchin major vault protein: a possible role for vault ribonucleoprotein particles in nucleocytoplasmic transport. AB - Vaults are large ribonucleoprotein particles that have been identified in a wide range of eukaryotic organisms. Although present in thousands of copies per cell, their function remains unknown. In this report, we identify the major vault protein in sea urchins as a 107-kDa polypeptide that copurifies with microtubules and ribosomes. Although initially identified in microtubule preparations, the sea urchin major vault protein is not predominantly microtubule-associated in vivo. Rather, the sea urchin major vault protein is present throughout the cytoplasm in eggs and embryos and in the nucleus in adult somatic cells. Within the nucleus, the sea urchin major vault protein is concentrated in the region of the nucleolus and to punctate regions of the nuclear envelope. In addition, the vault protein localizes to short linear strings juxtaposed to the exterior of the nucleus and extending outward into the cytoplasm. Based on their copurification and intracellular distribution, vaults may be involved in the nucleocytoplasmic transport of ribosomes and/or mRNA. PMID- 9331334 TI - Differential dependency of cutaneous mechanoreceptors on neurotrophins, trk receptors, and P75 LNGFR. AB - The impact of null mutations of the genes for the NGF family of neurotrophins and their receptors was examined among the wide variety of medium to large caliber myelinated mechanoreceptors which have a highly specific predictable organization in the mystacial pad of mice. Immunofluorescence with anti-protein gene product 9.5, anti-200-kDa neurofilament protein (RT97), and anti-calcitonin gene-related product was used to label innervation in mystacial pads from mice with homozygous null mutations for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), the three tyrosine kinase receptors (trkA, trkB, trkC), and the low-affinity nerve growth factor receptor p75. Specimens were sacrificed at birth and at 1, 2, and 4 weeks for each type of mutation as well as at 11 weeks and 1 year for p75 and trkC mutations, respectively. Our results demonstrate several major concepts about the role of neurotrophins in the development of cutaneous mechanoreceptors that are supplied by medium to large caliber myelinated afferents. First, each of the high-affinity tyrosine kinase receptors, trkA, trkB, and trkC, as well as the low-affinity p75 receptor has an impact on at least one type of mechanoreceptor. Second, consistent with the various affinities for particular trk receptors, the elimination of NGF, BDNF, and NT-3 has an impact comparable to or more complex than the absence of their most specific high-affinity receptors: trkA, trkB, and trkC, respectively. These complexities include potential NT-3 signaling through trkA and trkB to support some neuronal survival. Third, most types of afferents are dependent on a different combination of neurotrophins and receptors for their survival: reticular and transverse lanceolate afferents are dependent upon NT-3, NGF, and trkA; Ruffini afferents upon BDNF and trkB; longitudinal lanceolate afferents upon NGF, trkA, BDNF, and trkB; and Merkel afferents on NGF, trkA, NT 3, trkC, and p75. NT-4 has no obvious detrimental impact on the mechanoreceptor development in the presence of BDNF. Fourth, NT-4 and BDNF signaling through trkB may suppress Merkel innervation and NT-3 signaling through trkC may suppress Ruffini innervation. Finally, regardless of the neurotrophin/receptor dependency for afferent survival and neurite outgrowth, NT-3 has an impact on the formation of all the sensory endings. In the context of these findings, indications of competitive and suppressive interactions that appear to regulate the balance of innervation density among the various sets of innervation were evident. PMID- 9331336 TI - Arrowhead encodes a LIM homeodomain protein that distinguishes subsets of Drosophila imaginal cells. AB - The Arrowhead gene encodes a LIM-homeodomain transcription factor required for establishment of a subset of imaginal tissues: the abdominal histoblasts and the salivary gland imaginal rings. Consistent with its role in development, during embryogenesis Arrowhead is expressed in each abdominal segment and in the labial segment. Late in embryonic development, expression is refined to the abdominal histoblasts and salivary gland imaginal ring cells themselves. When ectopically expressed in imaginal disc cells, Arrowhead causes programmed cell death and loss of corresponding adult structures. Therefore, Arrowhead expression is required for development of one set of imaginal cells and is incompatible with development of another, emphasizing the specificity of Arrowhead and the sensitivity of different target cells to its expression. Loss-of-function mutations in Arrowhead affect conserved or invariant amino acids in the LIM- and homeo-domains demonstrating the importance of these residues in LIM homeodomain protein activity. PMID- 9331337 TI - Introduction of cyclin B induces activation of the maturation-promoting factor and breakdown of germinal vesicle in growing zebrafish oocytes unresponsive to the maturation-inducing hormone. AB - When treated with 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-DP), a natural maturation-inducing hormone in fishes, fully grown zebrafish oocytes are induced to mature via the activation of the maturation-promoting factor (MPF), which consists of cdc2 (a catalytic subunit) and cyclin B (a regulatory subunit). In contrast, 17alpha,20beta-DP is unable to induce growing (previtellogenic and vitellogenic) oocytes to mature. To know the reason growing oocytes fail to mature upon 17alpha,20beta-DP treatment, we investigated changes in the components of machinery responsible for MPF activation during zebrafish oogenesis. Immunoblotting experiments using monoclonal antibodies against cdc2, cyclin B, and cdk7 (an activator of cdc2) have revealed that the concentrations of cdc2 and cdk7 are almost constant during oogenesis. Cyclin B was present in mature oocytes but absent in growing and fully grown immature oocytes. These results, which are identical to those in goldfish, strongly suggest that cyclin B is synthesized from stored (masked) mRNA after 17alpha,20beta-DP stimulation and that its binding to the preexisting cdc2 allows cdk7 to activate MPF. Microinjection of cyclin B protein induced MPF activation and germinal vesicle breakdown in growing oocytes, as well as in fully grown oocytes, indicating that cdk7 present in growing oocytes is already active. Northern blot analysis revealed the presence of cyclin B mRNA in both previtellogenic and fully grown oocytes. These results indicate that, as in fully grown oocytes, growing oocytes are already equipped with the catalytic subunit of MPF (cdc2) and its activator (cdk7) and that the appearance of the regulatory subunit of MPF (cyclin B) is sufficient for initiating maturation. Therefore, the unresponsiveness of growing oocytes to 17alpha,20beta-DP is attributable to a deficiency in the processes leading to cyclin B synthesis, which include 17alpha,20beta-DP reception on the oocyte surface, subsequent signal transduction pathways, and unmasking the stored cyclin B mRNA. PMID- 9331338 TI - Chemical two-photon uncaging: a novel approach to mapping glutamate receptors. AB - Functional mapping of neurotransmitter receptors requires rapid and localized application of transmitter. The usefulness of caged glutamate for this purpose has been limited, because photolysis by unfocused light above and below the target cell limits depth resolution. This problem is eliminated by using a double caged glutamate that requires absorption of two photons for conversion to active glutamate, resulting in a substantial improvement in spatial resolution over conventional caged glutamate. This method was used to map the distribution of glutamate receptors on hippocampal pyramidal neurons. A higher density of AMPA receptors was found on distal apical dendrites than on basal or primary apical dendrites, suggesting that synaptic efficacy is locally heterogeneous. Such "chemical two-photon uncaging" offers a simple, general, and economical strategy for spatially localized photolysis of caged compounds. PMID- 9331339 TI - Silent synapses speak up. PMID- 9331340 TI - How does the brain smell? PMID- 9331341 TI - To cortex: thanks for the memories. PMID- 9331342 TI - Nomenclature of GPI-linked receptors for the GDNF ligand family. GFR(alpha) Nomenclature Committee. PMID- 9331343 TI - Assignment of early caudal identity to neural plate cells by a signal from caudal paraxial mesoderm. AB - The early patterning of the vertebrate central nervous system involves the generation of progenitor cells with distinct fates at rostral and caudal levels of the neuraxis. We provide evidence that the assignment of early rostrocaudal differences in progenitor cell properties is established by spatial restrictions in the signaling properties of the paraxial mesoderm and epidermal ectoderm. Caudal level paraxial mesoderm secretes a factor, distinct from retinoic acid or fibroblast growth factors (FGFs), that can impose caudal fates on prospective anterior proencephalic progenitors. The caudalizing activity of the paraxial mesoderm can, however, be induced by FGF signaling. The distinct properties of cells at rostral and caudal levels of the neural plate appear to depend, in addition, on the early exclusion of bone morphogenetic proteins (BMPs) from rostral level epidermal ectoderm. Thus, differences in the signaling properties of cell groups that flank the neural plate appear to contribute to the early rostrocaudal identity of neural cells, distinguishing progenitor cells at prospective anterior proencephalic regions from those at more caudal levels of the neuraxis. PMID- 9331344 TI - Introduction of a neurotrophin-3 transgene into muscle selectively rescues proprioceptive neurons in mice lacking endogenous neurotrophin-3. AB - To clarify the role of muscle-derived neurotrophin-3 (NT-3) in the development of sensory neurons, we generated transgenic mice selectively overexpressing NT-3 in skeletal muscles under the control of a myogenin promoter (myo-NT-3 mice). The myo-NT-3 transgene was then bred into an NT-3 null mutant (-/-) line to generate myo-NT-3, NT-3(-/-) mice in which NT-3 was expressed in muscles, but not elsewhere. Transient overexpression of NT-3 in developing muscles increased the number of proprioceptive neurons as well as the density of both their central and peripheral projections, resulting in more Ia afferents in spinal cord and more spindles (end organs of Ia afferents) in muscles. NT-3 expression restricted to muscles was sufficient to secure the development of proprioceptive neurons and their central and peripheral projections in myo-NT-3, NT-3(-/-) mice. The loss of nonproprioceptive neurons observed in NT-3(-/-) mice was not reversed by the transgene, suggesting that these neurons are regulated by NT-3 from sources other than muscle. We conclude that target-derived rather than intraganglionic NT-3 is preeminent in supporting the development of proprioceptive neurons. The level of NT-3 in developing muscles may be the principal factor determining the number of proprioceptive neurons in dorsal root ganglions and spindles in skeletal muscles of adults. PMID- 9331345 TI - Disruption of semaphorin III/D gene causes severe abnormality in peripheral nerve projection. AB - The molecules of the collapsin/semaphorin gene family have been thought to play an essential role in axon guidance during development. Semaphorin III/D is a member of this family, has been shown to repel dorsal root ganglion (DRG) axons in vitro, and has been implicated in the patterning of sensory afferents in the spinal cord. Although semaphorin III/D mRNA is expressed in a wide variety of neural and nonneural tissues in vivo, the role played by semaphorin III/D in regions other than the spinal cord is not known. Here, we show that mice homozygous for a targeted mutation in semaphorin III/D show severe abnormality in peripheral nerve projection. This abnormality is seen in the trigeminal, facial, vagus, accessory, and glossopharyngeal nerves but not in the oculomotor nerve. These results suggest that semaphorin III/D functions as a selective repellent in vivo. PMID- 9331346 TI - A 70 amino acid region within the semaphorin domain activates specific cellular response of semaphorin family members. AB - The semaphorin family contains secreted and transmembrane signaling proteins that function in the nervous, immune, and cardiovascular systems. Chick collapsin-1 is a repellent for specific growth cones. Two other secreted members of the semaphorin family, collapsin-2 and -3, are structurally similar to collapsin-1 but have different biological activities. Semaphorins contain a 500 amino acid family signature semaphorin domain. We show in this study that (1) the semaphorin domain of collapsin-1 is both necessary and sufficient for biological activity, (2) the semaphorin domain contains a 70 amino acid region that specifies the biological activity of the three family members, and (3) the positively charged carboxy terminus potentiates activity without affecting specificity. We propose that semaphorins interact with their receptors through two independent binding sites: one that mediates the biological response and one that potentiates it. PMID- 9331347 TI - Secreted chick semaphorins bind recombinant neuropilin with similar affinities but bind different subsets of neurons in situ. AB - Collapsin-1, a member of the semaphorin family, activates receptors on specific growth cones, thereby inhibiting their motility. Neuropilin, a previously cloned transmembrane protein, has recently been identified as a candidate receptor for collapsin-1. We have completed the cloning of chick collapsin-3 and -5 and show that collapsin-1, -2, -3, and -5 bind to overlapping but distinct axon tracts. We infer that in situ, there are distinct receptors with different affinities for collapsin-1, -2, -3, and -5. In contrast, these four collapsins all bind recombinant neuropilin with similar affinities. Strong binding to neuropilin is mediated by the carboxy third of the collapsins, while the semaphorin domain confers their unique binding patterns in situ. We propose that neuropilin is a common component of a semaphorin receptor complex, and that additional differentially expressed receptor components interact with the semaphorin domains to confer binding specificity. PMID- 9331349 TI - Genetic analysis of the mechanisms controlling target selection: target-derived Fasciclin II regulates the pattern of synapse formation. AB - In Drosophila, motoneuron growth cones initially probe many potential muscle targets but later withdraw most of these contacts to form stereotypic synapses with only one or a few muscles. Prior to synapse formation, Fasciclin II (Fas II) is expressed at low levels on muscle. During synapse formation, Fas II concentrates at the synapse and disappears from the rest of the muscle. We previously showed that Fas II is required both pre- and postsynaptically for synaptic stabilization. Here, we show that the differential expression of target derived Fas II has a profound influence on the patterning of synapse formation. A transient increase in muscle Fas II stabilizes growth cone contacts and leads to novel synapses that are functional and stable; targets that normally receive two inputs can now receive up to six inputs. Changing the relative levels of Fas II on neighboring muscles leads to dramatic shifts in target selection. PMID- 9331348 TI - Neuropilin-2, a novel member of the neuropilin family, is a high affinity receptor for the semaphorins Sema E and Sema IV but not Sema III. AB - Semaphorins are a large family of secreted and transmembrane proteins, several of which are implicated in repulsive axon guidance. Neuropilin (neuropilin-1) was recently identified as a receptor for Collapsin-1/Semaphorin III/D (Sema III). We report the identification of a related protein, neuropilin-2, whose mRNA is expressed by developing neurons in a pattern largely, though not completely, nonoverlapping with that of neuropilin-1. Unlike neuropilin-1, which binds with high affinity to the three structurally related semaphorins Sema III, Sema E, and Sema IV, neuropilin-2 shows high affinity binding only to Sema E and Sema IV, not Sema III. These results identify neuropilins as a family of receptors (or components of receptors) for at least one semaphorin subfamily. They also suggest that the specificity of action of different members of this subfamily may be determined by the complement of neuropilins expressed by responsive cells. PMID- 9331350 TI - Netrin-1 and DCC mediate axon guidance locally at the optic disc: loss of function leads to optic nerve hypoplasia. AB - Embryonic retinal ganglion cell (RGC) axons must extend toward and grow through the optic disc to exit the eye into the optic nerve. In the embryonic mouse eye, we found that immunoreactivity for the axon guidance molecule netrin-1 was specifically on neuroepithelial cells at the disk surrounding exiting RGC axons, and RGC axons express the netrin receptor, DCC (deleted in colorectal cancer). In vitro, anti-DCC antibodies reduced RGC neurite outgrowth responses to netrin-1. In netrin-1- and DCC-deficient embryos, RGC axon pathfinding to the disc was unaffected; however, axons failed to exit into the optic nerve, resulting in optic nerve hypoplasia. Thus, netrin-1 through DCC appears to guide RGC axons locally at the optic disc rather than at long range, apparently reflecting the localization of netrin-1 protein to the vicinity of netrin-1-producing cells at the optic disc. PMID- 9331351 TI - Acute effects of cocaine on human brain activity and emotion. AB - We investigated brain circuitry mediating cocaine-induced euphoria and craving using functional MRI (fMRI). During double-blind cocaine (0.6 mg/kg) and saline infusions in cocaine-dependent subjects, the entire brain was imaged for 5 min before and 13 min after infusion while subjects rated scales for rush, high, low, and craving. Cocaine induced focal signal increases in nucleus accumbens/subcallosal cortex (NAc/SCC), caudate, putamen, basal forebrain, thalamus, insula, hippocampus, parahippocampal gyrus, cingulate, lateral prefrontal and temporal cortices, parietal cortex, striate/extrastriate cortices, ventral tegmentum, and pons and produced signal decreases in amygdala, temporal pole, and medial frontal cortex. Saline produced few positive or negative activations, which were localized to lateral prefrontal cortex and temporo occipital cortex. Subjects who underwent repeat studies showed good replication of the regional fMRI activation pattern following cocaine and saline infusions, with activations on saline retest that might reflect expectancy. Brain regions that exhibited early and short duration signal maxima showed a higher correlation with rush ratings. These included the ventral tegmentum, pons, basal forebrain, caudate, cingulate, and most regions of lateral prefrontal cortex. In contrast, regions that demonstrated early but sustained signal maxima were more correlated with craving than with rush ratings; such regions included the NAc/SCC, right parahippocampal gyrus, and some regions of lateral prefrontal cortex. Sustained negative signal change was noted in the amygdala, which correlated with craving ratings. Our data demonstrate the ability of fMRI to map dynamic patterns of brain activation following cocaine infusion in cocaine-dependent subjects and provide evidence of dynamically changing brain networks associated with cocaine induced euphoria and cocaine-induced craving. PMID- 9331352 TI - Fear conditioning enhances different temporal components of tone-evoked spike trains in auditory cortex and lateral amygdala. AB - Single neurons were recorded in freely behaving rats during fear conditioning from areas of auditory cortex that project to the lateral nucleus of the amygdala (LA). The latency and rate of conditioning and extinction were analyzed, and the results were compared to previous recordings from LA itself. Auditory cortex neurons took more trials to learn, and they responded more slowly than LA neurons within trials. Short-latency plasticity in LA, therefore, reflects inputs from the auditory thalamus rather than the auditory cortex. Unlike LA cells, some auditory cortex cells showed late conditioned responses that seemed to anticipate the unconditioned stimulus, while others showed extinction-resistant memory storage. Thus, rapid conditioning of fear responses to potentially dangerous stimuli depends on plasticity in the amygdala, while cortical areas may be particularly involved in higher cognitive (mnemonic and attentional) processing of fear experiences. PMID- 9331353 TI - Regulation of dendritic growth and remodeling by Rho, Rac, and Cdc42. AB - The acquisition of cell type-specific morphologies is a central feature of neuronal differentiation and has important consequences for nervous system function. To begin to identify the underlying molecular mechanisms, we have explored the role of Rho-related GTPases in the dendritic development of cortical neurons. Expression of dominant negative mutants of Rac or Cdc42, the Rho inhibitory molecule C3 transferase, or the GTPase-activating protein RhoGAP p190 causes a marked reduction in the number of primary dendrites in nonpyramidal (multipolar) neurons and in the number of basal dendrites in neurons with pyramidal morphologies. Conversely, the expression of constitutively active mutants of Rho, Rac, or Cdc42 leads to an increase in the number of primary and basal dendrites. In cortical cultures, as in vivo, dendritic remodeling leads to an apparent transformation from pyramidal to nonpyramidal morphologies over time. Strikingly, this shift in favor of nonpyramidal morphologies is also inhibited by the expression of dominant negative mutants of Cdc42 and Rac and by RhoGAP p190. These observations indicate that Rho, Rac, and Cdc42 play a central role in dendritic development and suggest that differential activation of Rho-related GTPases may contribute to the generation of morphological diversity in the developing cortex. PMID- 9331354 TI - Recruitment of new sites of synaptic transmission during the cAMP-dependent late phase of LTP at CA3-CA1 synapses in the hippocampus. AB - Long-term potentiation at CA3-CA1 hippocampal synapses exhibits an early phase and a late phase, which can be distinguished by their underlying molecular mechanisms. Unlike the early phase, the late phase is dependent on both cAMP and protein synthesis. Quantal analysis of unitary synaptic transmission between a single presynaptic CA3 neuron and a single postsynaptic CA1 neuron suggests that, under certain conditions, the early phase of LTP involves an increase in the probability of release of a single quantum of transmitter from a single presynaptic release site, with no change in the number of quanta that are released or in postsynaptic sensitivity to transmitter. Here, we show that the cAMP-induced late phase of LTP involves an increase in the number of quanta released in response to a single presynaptic action potential, possibly due to an increase in the number of sites of synaptic transmission between a single CA3 and a single CA1 neuron. PMID- 9331355 TI - Neurotrophins and time: different roles for TrkB signaling in hippocampal long term potentiation. AB - We examined the role of TrkB ligands in hippocampal long-term potentiation (LTP) using function-blocking TrkB antiserum (Ab) and Trk-IgG fusion proteins. Incubation of hippocampal slices with TrkB Ab had no effect on basal synaptic transmission, short-term plasticity, or LTP induced by several trains of tetanic stimulation. The TrkB Ab-treated slices, however, showed significant deficits in LTP induced by either theta-burst stimulation (TBS) or "pairing." Slices exposed to the same number of inducing stimuli, delivered either as TBS or as a single 100 Hz epoch, only exhibited TrkB-sensitive LTP when TBS was used, indicating that the temporal pattern of stimulation determines the neurotrophin dependence. The late phase of LTP (2-3 hr) was also significantly impaired in slices pretreated with TrkB Ab or a TrkB-IgG. The application of a TrkB-IgG 30 min after LTP induction caused previously potentiated synaptic transmission to return to baseline levels, indicating that TrkB ligands are required to maintain LTP for up to 1 hr after induction. Taken together, these results indicate that both the temporal patterns of synaptic activity and the different temporal phases of synaptic enhancement are important in determining the neurotrophin dependence of plasticity in the hippocampus. PMID- 9331356 TI - Tonic synaptic inhibition modulates neuronal output pattern and spatiotemporal synaptic integration. AB - Irregular firing patterns are observed in most central neurons in vivo, but their origin is controversial. Here, we show that two types of inhibitory neurons in the cerebellar cortex fire spontaneously and regularly in the absence of synaptic input but generate an irregular firing pattern in the presence of tonic synaptic inhibition. Paired recordings between synaptically connected neurons revealed that single action potentials in inhibitory interneurons cause highly variable delays in action potential firing in their postsynaptic cells. Activity in single and multiple inhibitory interneurons also significantly reduces postsynaptic membrane time constant and input resistance. These findings suggest that the time window for synaptic integration is a dynamic variable modulated by the level of tonic inhibition, and that rate coding and temporal coding strategies may be used in parallel in the same cell type. PMID- 9331357 TI - Differential regulation of neocortical synapses by neuromodulators and activity. AB - Synapses are continually regulated by chemical modulators and by their own activity. We tested the specificity of regulation in two excitatory pathways of the neocortex: thalamocortical (TC) synapses, which mediate specific inputs, and intracortical (IC) synapses, which mediate the recombination of cortical information. Frequency-sensitive depression was much stronger in TC synapses than in IC synapses. The two synapse types were differentially sensitive to presynaptic neuromodulators: only IC synapses were suppressed by activation of GABA(B) receptors, only TC synapses were enhanced by nicotinic acetylcholine receptors, and muscarinic acetylcholine receptors suppressed both synapse types. Modulators also differentially altered the frequency sensitivity of the synapses. Our results suggest a mechanism by which the relative strength and dynamics of input and associational pathways of neocortex are regulated during changes in behavioral state. PMID- 9331358 TI - G protein-coupled inwardly rectifying K+ channels (GIRKs) mediate postsynaptic but not presynaptic transmitter actions in hippocampal neurons. AB - To study the role of G protein-coupled, inwardly rectifying K+ (GIRK) channels in mediating neurotransmitter actions in hippocampal neurons, we have examined slices from transgenic mice lacking the GIRK2 gene. The outward currents evoked by agonists for GABA(B) receptors, 5HT1A receptors, and adenosine A1 receptors were essentially absent in mutant mice, while the inward current evoked by muscarinic receptor activation was unaltered. In contrast, the presynaptic inhibitory action of a number of presynaptic receptors on excitatory and inhibitory terminals was unaltered in mutant mice. These included GABA(B), adenosine, muscarinic, metabotropic glutamate, and NPY receptors on excitatory synapses and GABA(B) and opioid receptors on inhibitory synapses. These findings suggest that a number of G protein-coupled receptors activate the same class of postsynaptic K+ channel, which contains GIRK2. In addition, the GIRK2 channels play no role in the inhibition mediated by presynaptic G protein-coupled receptors, suggesting that the same receptor can couple to different effector systems according to its subcellular location in the neuron. PMID- 9331359 TI - Differences in synaptic GABA(A) receptor number underlie variation in GABA mini amplitude. AB - In many neurons, responses to individual quanta of transmitter exhibit large variations in amplitude. The origin of this variability, although central to our understanding of synaptic transmission and plasticity, remains controversial. To examine the relationship between quantal amplitude and postsynaptic receptor number, we adopted a novel approach, combining patch-clamp recording of synaptic currents with quantitative immunogold localization of synaptic receptors. Here, we report that in cerebellar stellate cells, where variability in GABA miniature synaptic currents is particularly marked, the distribution of quantal amplitudes parallels that of synaptic GABA(A) receptor number. We also show that postsynaptic GABA(A) receptor density is uniform, allowing synaptic area to be used as a measure of relative receptor content. Flurazepam, which increases GABA(A) receptor affinity, prolongs the decay of all miniature currents but selectively increases the amplitude of large events. From this differential effect, we show that a quantum of GABA saturates postsynaptic receptors when <80 receptors are present but results in incomplete occupancy at larger synapses. PMID- 9331360 TI - RNA editing generates a diverse array of transcripts encoding squid Kv2 K+ channels with altered functional properties. AB - We have cloned a Kv2 potassium channel from squid optic lobe termed sqKv2. Multiple overlapping sqKv2 cDNA clones differed from one another at specific positions by purine transitions. To test whether the purine transitions were generated by RNA editing, we compared a 360 nucleotide genomic sequence with corresponding cDNA sequences (encoding S4-S6) isolated from individual animals and lying on a single gene and exon. cDNA sequences differed from genomic sequence at 17 positions, resulting in 28 unique sequences. There was invariantly an adenosine in the genomic sequence and a guanosine in the edited cDNA sequences. Two of the edits altered the rates of channel closure and slow inactivation. These results extend selective RNA editing to invertebrate taxa and represents a novel mechanism for the posttranscriptional modulation of voltage gated ion channels. PMID- 9331361 TI - Xestospongins: potent membrane permeable blockers of the inositol 1,4,5 trisphosphate receptor. AB - Xestospongins (Xe's) A, C, D, araguspongine B, and demethylxestospongin B, a group of macrocyclic bis-1-oxaquinolizidines isolated from the Australian sponge, Xestospongia species, are shown to be potent blockers of IP3-mediated Ca2+ release from endoplasmic reticulum vesicles of rabbit cerebellum. XeC blocks IP3 induced Ca2+ release (IC50 = 358 nM) without interacting with the IP3-binding site, suggesting a mechanism that is independent of the IP3 effector site. Analysis of Pheochromocytoma cells and primary astrocytes loaded with Ca2+ sensitive dye reveals that XeC selectively blocks bradykinin- and carbamylcholine induced Ca2+ efflux from endoplasmic reticulum stores. Xe's represent a new class of potent, membrane permeable IP3 receptor blockers exhibiting a high selectivity over ryanodine receptors. Xe's are a valuable tool for investigating the structure and function of IP3 receptors and Ca2+ signaling in neuronal and nonneuronal cells. PMID- 9331362 TI - When the chips are down. PMID- 9331363 TI - Modeling human evolution--to tree or not to tree? PMID- 9331364 TI - Gaucher disease phenotypes outflanked? PMID- 9331365 TI - Database divisions and homology search files: a guide for the perplexed. PMID- 9331366 TI - Long human-mouse sequence alignments reveal novel regulatory elements: a reason to sequence the mouse genome. PMID- 9331367 TI - The role of genomics in studying genetic susceptibility to infectious disease. PMID- 9331368 TI - Interpreting a sequenced genome: toward a cosmid transgenic library of Caenorhabditis elegans. AB - We have generated a library of transgenic Caenorhabditis elegans strains that carry sequenced cosmids from the genome of the nematode. Each strain carries an extrachromosomal array containing a single cosmid, sequenced by the C. elegans Genome Sequencing Consortium, and a dominate Rol-6 marker. More than 500 transgenic strains representing 250 cosmids have been constructed. Collectively, these strains contain approximately 8 Mb of sequence data, or approximately 8% of the C. elegans genome. The transgenic strains are being used to rescue mutant phenotypes, resulting in a high-resolution map alignment of the genetic, physical, and DNA sequence maps of the nematode. We have chosen the region of chromosome III deleted by sDf127 and not covered by the duplication sDp8(III;I) as a starting point for a systematic correlation of mutant phenotypes with nucleotide sequence. In this defined region, we have identified 10 new essential genes whose mutant phenotypes range from developmental arrest at early larva, to maternal effect lethal. To date, 8 of these 10 essential genes have been rescued. In this region, these rescues represent approximately 10% of the genes predicted by GENEFINDER and considerably enhance the map alignment. Furthermore, this alignment facilitates future efforts to physically position and clone other genes in the region. [Updated information about the Transgenic Library is available via the Internet at http://darwin.mbb.sfu.ca/imbb/dbaillie/cos mid.html.] PMID- 9331369 TI - The significance of digital gene expression profiles. AB - Genes differentially expressed in different tissues, during development, or during specific pathologies are of foremost interest to both basic and pharmaceutical research. "Transcript profiles" or "digital Northerns" are generated routinely by partially sequencing thousands of randomly selected clones from relevant cDNA libraries. Differentially expressed genes can then be detected from variations in the counts of their cognate sequence tags. Here we present the first systematic study on the influence of random fluctuations and sampling size on the reliability of this kind of data. We establish a rigorous significance test and demonstrate its use on publicly available transcript profiles. The theory links the threshold of selection of putatively regulated genes (e.g., the number of pharmaceutical leads) to the fraction of false positive clones one is willing to risk. Our results delineate more precisely and extend the limits within which digital Northern data can be used. PMID- 9331370 TI - Detection of numerous Y chromosome biallelic polymorphisms by denaturing high performance liquid chromatography. AB - Y chromosome haplotypes are particularly useful in deciphering human evolutionary history because they accentuate the effects of drift, migration, and range expansion. Significant acceleration of Y biallelic marker discovery and subsequent typing involving heteroduplex detection has been achieved by implementing an innovative and cost-efficient method called denaturing high performance liquid chromatography (DHPLC). The power of the method resides in its sensitivity and ability to rapidly compare amplified sequences in an automated manner. We have determined the allelic states of 22 Y polymorphisms; 19 of which are unreported, in 718 diverse extant chromosomes; established haplotype frequencies; and deduced a phylogeny. All major geographic regions, including Eurasia, are characterized by mutations reflecting episodes of genetic drift and expansion. Most biallelic markers are localized regionally. However, some show wider dispersal and designate older, core haplotypes. One transversion defines a major haplogroup that distinguishes a previously unknown deep, apparently non African branch. It provides evidence of an ancient bottleneck event. It is now possible to anticipate the inevitable detailed reconstruction of human Y chromosome genealogy based on several tens to even hundreds of these important polymorphisms. PMID- 9331373 TI - IMAGE cDNA clones, UniGene clustering, and ACeDB: an integrated resource for expressed sequence information. AB - In this study we describe a new information resource that provides integrated access to information on IMAGE (integrated molecular analysis of genomes and their expression) cDNA library clones and derived expressed sequence tags (ESTs). We have developed an automated procedure that collates data from various public sources into a single ACeDB database. This database is a valuable tool for electronic cloning experiments and gene expression studies. It allows researchers to find information about cDNA libraries, plate addresses, insert sizes, and sequence data for IMAGE clones, the assignment of ESTs to UniGene clusters, and the chromosomal location of those genes in an efficient, graphically oriented manner. PMID- 9331372 TI - Identification of three additional genes contiguous to the glucocerebrosidase locus on chromosome 1q21: implications for Gaucher disease. AB - Gaucher disease results from the deficiency of the lysosomal enzyme glucocerebrosidase (EC 3.2.1.45). Although the functional gene for glucocerebrosidase (GBA) and its pseudogene (psGBA), located in close proximity on chromosome 1q21, have been studied extensively, the flanking sequence has not been well characterized. The recent identification of human metaxin (MTX) immediately downstream of psGBA prompted a closer analysis of the sequence of the entire region surrounding the GBA gene. We now report the genomic DNA sequence and organization of a 75-kb region around GBA, including the duplicated region containing GBA and MTX. The origin and endpoints of the duplication leading to the pseudogenes for GBA and MTX are now clearly established. We also have identified three new genes within the 32 kb of sequence upstream to GBA, all of which are transcribed in the same direction as GBA. Of these three genes, the gene most distal to GBA is a protein kinase (clk2). The second gene, propin1, has a 1.5-kb cDNA and shares homology to a rat secretory carrier membrane protein 37 (SCAMP37). Finally, cote1, a gene of unknown function lies most proximal to GBA. The possible contributions of these closely arrayed genes to the more atypical presentations of Gaucher disease is now under investigation. PMID- 9331371 TI - The largest subunit of human RNA polymerase III is closely related to the largest subunit of yeast and trypanosome RNA polymerase III. AB - In both yeast and mammalian systems, considerable progress has been made toward the characterization of the transcription factors required for transcription by RNA polymerase III. However, whereas in yeast all of the RNA polymerase III subunits have been cloned, relatively little is known about the enzyme itself in higher eukaryotes. For example, no higher eukaryotic sequence corresponding to the largest RNA polymerase III subunit is available. Here we describe the isolation of cDNAs that encode the largest subunit of human RNA polymerase III, as suggested by the observations that (1) antibodies directed against the cloned protein immunoprecipitate an active enzyme whose sensitivity to different concentrations of alpha-amanitin is that expected for human RNA polymerase III; and (2) depletion of transcription extracts with the same antibodies results in inhibition of transcription from an RNA polymerase III, but not from an RNA polymerase II, promoter. Sequence comparisons reveal that regions conserved in the RNA polymerase I, II, and III largest subunits characterized so far are also conserved in the human RNA polymerase III sequence, and thus probably perform similar functions for the human RNA polymerase III enzyme. PMID- 9331374 TI - Sarcopenia and physical performance in old age: overview. PMID- 9331375 TI - Sarcopenia and physical performance in old age: introduction. PMID- 9331376 TI - Assessment of physical performance and disability in older persons. PMID- 9331377 TI - Quantifying physical functional performance in older adults. PMID- 9331378 TI - Targeting disabled and at risk older adults for inclusion in population based studies. PMID- 9331379 TI - Inclusion of ethnic minorities in sarcopenia research: challenges and strategies. PMID- 9331380 TI - Statistical principles underlying the applications of laboratory methods to field studies. PMID- 9331381 TI - Confounding variables and comorbidity in sarcopenia research. PMID- 9331382 TI - Functional and behavioral consequences of sarcopenia. PMID- 9331383 TI - Measurement of muscle strength and power. PMID- 9331384 TI - Choosing the best strength measure in frail older persons: importance of task specificity. PMID- 9331385 TI - How do the elderly negotiate stairs? AB - Stair navigation, particularly stair descent, is an extremely challenging and dangerous locomotor task, yet studies suggest that most elderly are unlikely to move to new residences in order to avoid this challenge. The knee and ankle are the key joints where adequate strength and power are required for safe stair descent, and it is not yet clear if sarcopenia in the elderly is likely to result in residual strength below that which is required for successful stair performance. Sensory cues are also critical, and the lack of literature on the specific roles of the various intrinsic and extrinsic factors that affect stair navigation is a clear indication of the need for such research in order to define safer strategies and optimal conditions for elderly individuals to transit between living areas of differing levels. PMID- 9331386 TI - Muscle strength and rising from a chair in older adults. PMID- 9331388 TI - Neural strategies in the control of muscle force. PMID- 9331387 TI - What leads to age and gender differences in balance maintenance and recovery? AB - OA compared to YA have high rates of falls and fall-related injuries. OF have notably higher rates of falls and fall-related injuries than OM. Healthy OA compared to YA, and females compared to males of any adult age, have lower strengths and have development rates for at least some strengths that are lower. The results of the obstacle avoidance and balance recovery studies described suggest that OA are not notably more at risk than YA, nor are females notably more at risk than males, in avoidance and recovery tasks that are time-critical (TC), but do not have high strength (HS) requirements. The results suggest that for TC/HS avoidance and recovery tasks, OA compared to YA and females compared to males are substantially more at risk for injury. The source of these age and gender differences seems to lie primarily in differences in muscle strengths and speeds of muscle contraction once contraction is initiated, rather than in neural factors underlying the sensory processing or motor planning that leads to the initiation of muscle contraction. Perhaps these findings help to explain the high rates of falls and fall injuries among OA compared to YA, and among OF compared to OM. PMID- 9331389 TI - Quantitative methods for estimating the number of motor units in human muscles. PMID- 9331390 TI - Age-related changes in contractile properties and expression of myosin isoforms in single skeletal muscle cells. PMID- 9331391 TI - Analysis of apoptosis in culture models and intact tissues. PMID- 9331392 TI - Age-related changes in neuromuscular innervation. PMID- 9331393 TI - Excitation-Ca2+ release-contraction coupling in single aged human skeletal muscle fiber. PMID- 9331394 TI - Measurement of synthesis rates of specific muscle proteins using needle biopsy samples. PMID- 9331395 TI - Noninvasive measures of central and peripheral activation in human muscle fatigue. PMID- 9331396 TI - Magnetic resonance as a tool to study sarcopenia. PMID- 9331397 TI - Blood flow and substrate exchange in skeletal muscle of man: techniques relevant for use in the study of the ageing process of muscle. PMID- 9331398 TI - Muscle capillarization: morphological and morphometrical analyses of biopsy samples. PMID- 9331399 TI - Summary of work group I: population-based studies. PMID- 9331400 TI - Summary of work group II: small-scale clinical studies. PMID- 9331401 TI - Cell-to-cell contact and extracellular matrix. PMID- 9331403 TI - How corrinoids are synthesized without oxygen: nature's first pathway to vitamin B12. AB - BACKGROUND: During the biosynthesis of vitamin B12, the aerobic bacterium Pseudomonas denitrificans uses two enzymes, CobG and CobJ, to convert precorrin-3 to the ring-contracted intermediate, precorrin-4. CobG is a monooxygenase that adds a hydroxyl group, derived from molecular oxygen, to C-20, whereas CobJ is bifunctional, inserting a methyl group at C-17 of the macrocycle and catalyzing ring contraction. Molecular oxygen is not available to vitamin B12-producing anaerobic bacteria and members of the ancient Archaea, so the question arises of how these microbes accomplish the key ring-contraction process. RESULTS: Cloning and overexpression of Salmonella typhimurium genes has led to the discovery that a single enzyme, CbiH, is responsible for ring contraction during anaerobic biosynthesis of vitamin B12. The process occurs when CbiH is incubated with precorrin-3, but only in the presence of cobalt. CbiH functions as a C-17 methyltransferase and mediates ring contraction and lactonization to yield the intermediate, cobalt-precorrin-4, isolated as cobalt-factor IV. 13C labeling studies have proved that cobalt-precorrin-4 is incorporated into cobyrinic acid, thereby confirming that cobalt-precorrin-4 is an intermediate in vitamin B12 biosynthesis. CONCLUSIONS: Two distinct mechanisms exist in nature for the ring contraction of porphyrinoids to corrinoids-an ancient anaerobic pathway that requires cobalt complexation prior to nonoxidative rearrangement, and a more recent aerobic route in which molecular oxygen serves as the cofactor. The present results offer a rationale for the main differences between aerobic and anaerobic biosynthesis of vitamin B12. Thus, in anaerobes there is exchange of oxygen at the C-27 acetate site, extrusion of acetaldehyde and early insertion of cobalt, whereas the aerobes show no exchange of oxygen at C-27, extrude acetic acid and insert cobalt very late in the biosynthetic pathway, after ring contraction has occurred. These parallel routes to vitamin B12 have now been clearly distinguished by their differing mechanisms for ring contraction. PMID- 9331404 TI - Quantitative electrospray mass spectrometry for the rapid assay of enzyme inhibitors. AB - BACKGROUND: Combinatorial chemistry has become an important method for identifying effective ligand-receptor binding, new catalysts and enzyme inhibitors. In order to distinguish the most active component of a library or to obtain structure-activity relationships of compounds in a library, an efficient quantitative assay is crucial. Electrospray mass spectrometry has become an indispensable tool for qualitatively screening combinatorial libraries and its use for quantitative analysis has recently been demonstrated. RESULTS: This paper describes the use of quantitative electrospray mass spectrometry for screening libraries of inhibitors of enzymatic reactions, specifically the enzymatic glycosylation by beta-1,4-galactosyltransferase, which catalyzes the transfer of galactose from uridine-5'-diphosphogalactose to the 4-position of N acetylglucosamine beta OBn (Bn: benzene) to form N-acetyllactosamine beta OBn. Our mass spectrometric screening approach showed that both nucleoside diphosphates and triphosphates inhibited galactosyltransferase while none of the nucleoside monophosphates, including uridine-5'-monophosphate, showed any inhibition. Additional libraries were generated in which the concentrations of the inhibitors were varied and, using mass spectrometry, uridine-5'-diphosphate-2 deoxy-2-fluorogalactose was identified as the best inhibitor. CONCLUSIONS: This report introduces quantitative electrospray mass spectrometry as a rapid, sensitive and accurate quantitative assaying tool for inhibitor libraries that does not require a chromophore or radiolabeling. A viable alternative to existing analytical techniques is thus provided. The new technique will greatly facilitate the discovery of novel inhibitors against galactosyltransferase, an enzyme for which there are few potent inhibitors. PMID- 9331405 TI - Bacterial protein secretion--a target for new antibiotics? AB - The heavy use of antibiotics over recent decades has resulted in widespread resistance of bacteria to many drugs. Overcoming resistance requires new approaches to antibiotic development, including the exploitation of new targets in the bacterial cell. Protein secretion is essential for bacterial cell growth and virulence, so it could be a suitable target for new therapeutic agents. PMID- 9331406 TI - DNA recognition and bending. AB - DNA-binding proteins recognize their DNA targets not only through the formation of specific contacts with the nucleotide bases but also through inherent properties of the DNA sequence, including increased bendability and rigidity. Consideration of the properties of both the protein and the DNA is required before the sequence specificity and the observed DNA bend in DNA-protein complexes can be understood. PMID- 9331408 TI - Selection of RNA amide synthases. AB - BACKGROUND: It is generally accepted that, during evolution, replicating RNA molecules emerged from pools of random polynucleotides. This prebiotic RNA world was followed by an era of RNA-mediated catalysis of amide-bond formation. RNA would thus have provided the machinery responsible for the assembly of peptides and the beginning of the protein world of today. Naturally occurring ribozymes, which catalyze the cleavage or ligation of oligonucleotide phosphodiester bonds, support the idea that RNA could self-replicate. But was RNA constrained to this path and were RNA-acylated carriers required before RNA could catalyze the formation of amide bonds? RESULTS: We have isolated RNA catalysts that are capable of mediating amide-bond synthesis without the need for specifically designed templates to align the substrates, and we have kinetically characterized these catalysts. The rate enhancement observed for these RNA amide synthases exceeds the noncatalyzed amidation rate by a factor of approximately 10(4). In addition, Cu2+ ions caused a change in the affinity of RNA for the substrate rather than being directly involved in amide-bond formation. CONCLUSIONS: The discovery of these new amide synthases shows how functionally modified nucleic acids can facilitate covalent-bond formation without templating. Previously unforeseen RNA-evolution pathways can, therefore, be considered; for example, to guide amide-bond formation, en route to the protein world, it appears that substrate-binding pockets were formed that are analogous to those of protein enzymes. PMID- 9331407 TI - Engineered intermodular and intramodular polyketide synthase fusions. AB - BACKGROUND: Modular polyketide synthases (PKSs) are very large multifunctional enzyme complexes that synthesize a number of medicinally important natural products. The modular arrangement of active sites has made these enzyme systems amenable to combinatorial manipulation for the biosynthesis of novel polyketides. Here, we investigate the involvement of subunit interactions in hybrid and artificially linked PKSs with several series of intermodular and intramodular fusions using the erythromycin (6-deoxyerythronolide B synthase; DEBS) and rapamycin (RAPS) PKSs. RESULTS: Several two-module and three-module derivatives of DEBS were constructed by fusing module 6 to either module 2 or module 3 at varying junctions. Polyketide production by these intramodular fusions indicated that the core set of active sites remained functional in these hybrid modules, although the ketoreductase domain of module 6 was unable to recognize unnatural triketide and tetraketide substrates. Artificial trimodular PKS subunits were also engineered by covalently linking modules 2 and 3 of DEBS, thereby demonstrating the feasibility of constructing single-chain PKSs. Finally, a series of fusions containing DEBS and RAPS domains in module 2 of an engineered trimodular PKS revealed the structural and functional tolerance for hybrid modules created from distinct PKS gene clusters. CONCLUSIONS: The general success of the intermodular and intramodular fusions described here demonstrates significant structural tolerance among PKS modules and subunits and suggests that substrate specificity, rather than protein-protein interactions, is the primary determinant of molecular recognition features of PKSs. Furthermore, the ability to artificially link modules may considerably simplify the heterologous expression of modular PKSs in higher eukaryotic systems. PMID- 9331410 TI - Catalyst - for change in book reviews. PMID- 9331409 TI - Characterization of an 'orthogonal' suppressor tRNA derived from E. coli tRNA2(Gln). AB - BACKGROUND: In an effort to expand further our ability to manipulate protein structure, we have completed the first step towards a general method that allows the site-specific incorporation of unnatural amino acids into proteins in vivo. Our approach involves the construction of an 'orthogonal' suppressor tRNA that is uniquely acylated in vivo, by an engineered aminoacyl-tRNA synthetase, with the desired unnatural amino acid. The Escherichia coli tRNA2(Gln)-glutaminyl-tRNA synthetase (GlnRS) pair provides a biochemically and structurally well characterized starting point for developing this methodology. To generate the orthogonal tRNA, mutations were introduced into the acceptor stem, D-loop/stem, and anticodon loop of tRNA2(Gln). We report here the characterization of the properties of the resulting tRNAs and their suitability to severe as an orthogonal suppressor. Our efforts to generate an engineered synthetase are described elsewhere. RESULTS: Mutant tRNAs were generated by runoff transcription and assayed for their ability to be aminoacylated by purified E. coli GlnRS and to suppress an amber codon in an in vitro transcription/translation reaction. One tRNA bearing eight mutations satisfies the minimal requirements for the delivery of an unnatural amino acid: it is not acylated by any endogenous E. coli aminoacyl-tRNA synthetase, including GlnRS, yet functions efficiently during protein translation. Mutations in the acceptor stem and D-loop/stem, when introduced in combination, had very different effects on the properties of the resulting tRNAs compared with the effects of the individual mutations. CONCLUSIONS: Mutations at sites within tRNA2(Gln) separated by 23-31 A interact strongly with each other, often in a nonadditive fashion, to modulate both aminoacylation activities and translational efficiencies. The observed correlation between the effects of mutations at very distinct regions of the GlnRS-tRNA and possibly the ribosomal/tRNA complexes may contribute in part to the fidelity of protein biosynthesis. PMID- 9331411 TI - The critical active-site amine of the human 8-oxoguanine DNA glycosylase, hOgg1: direct identification, ablation and chemical reconstitution. AB - BACKGROUND: Base-excision DNA repair (BER) is the principal pathway responsible for the removal of aberrant, genotoxic bases from the genome and restoration of the original sequence. Key components of the BER pathway are DNA glycosylases, enzymes that recognize aberrant bases in the genome and catalyze their expulsion. One major class of such enzymes, glycosylase/lyases, also catalyze scission of the DNA backbone following base expulsion. Recent studies indicate that the glycosylase and lyase functions of these enzymes are mechanistically unified through a common amine-bearing residue on the enzyme, which acts as both the electrophile that displaces the aberrant base and an electron sink that facilitates DNA strand scission through imine (Schiff base)/conjugate elimination chemistry. The identity of this critical amine-bearing residue has not been rigorously established for any member of a superfamily of BER glycosylase/lyases. RESULTS: Here, we report the identification of the active-site amine of the human 8-oxoguanine DNA glycosylase (hOgg1), a human BER superfamily protein that repairs the mutagenic 8-oxoguanine lesion in DNA. We employed Edman sequencing of an active-site peptide irreversibly linked to substrate DNA to identify directly the active-site amine of hOgg1 as the epsilon-NH2 group of Lys249. In addition, we observed that the repair-inactive but recognition-competent Cys249 mutant (Lys249-->Cys) of hOgg1 can be functionally rescued by alkylation with 2 bromoethylamine, which functionally replaces the lysine residue by generating a gamma-thia-lysine. CONCLUSIONS: This study provides the first direct identification of the active-site amine for any DNA glycosylase/lyase belonging to the BER superfamily, members of which are characterized by the presence of a helix-hairpin-helix-Gly/Pro-Asp active-site motif. The critical lysine residue identified here is conserved in all members of the BER superfamily that exhibit robust glycosylase/lyase activity. The ability to trigger the catalytic activity of the Lys249-->Cys mutant of hOgg1 by treatment with the chemical inducer 2 bromoethylamine may permit snapshots to be taken of the enzyme acting on its substrate and could represent a novel strategy for conditional activation of catalysis by hOgg1 in cells. PMID- 9331412 TI - Unforeseen developments. PMID- 9331414 TI - Solution structure and base pair opening kinetics of the i-motif dimer of d(5mCCTTTACC): a noncanonical structure with possible roles in chromosome stability. AB - BACKGROUND: Repetitive cytosine-rich DNA sequences have been identified in telomeres and centromeres of eukaryotic chromosomes. These sequences play a role in maintaining chromosome stability during replication and may be involved in chromosome pairing during meiosis. The C-rich repeats can fold into an 'i-motif' structure, in which two parallel-stranded duplexes with hemiprotonated C.C+ pairs are intercalated. Previous NMR studies of naturally occurring repeats have produced poor NMR spectra. This led us to investigate oligonucleotides, based on natural sequences, to produce higher quality spectra and thus provide further information as to the structure and possible biological function of the i-motif. RESULTS: NMR spectroscopy has shown that d(5mCCTTTACC) forms an i-motif dimer of symmetry-related and intercalated folded strands. The high-definition structure is computed on the basis of the build-up rates of 29 intraresidue and 35 interresidue nuclear Overhauser effect (NOE) connectivities. The i-motif core includes intercalated interstrand C.C+ pairs stacked in the order 2*.8/1.7*/1*.7/2.8* (where one strand is distinguished by an asterisk and the numbers relate to the base positions within the repeat). The TTTA sequences form two loops which span the two wide grooves on opposite sides of the i-motif core; the i-motif core is extended at both ends by the stacking of A6 onto C2.C8+. The lifetimes of pairs C2.C8+ and 5mC1.C7+ are 1 ms and 1 s, respectively, at 15 degrees C. Anomalous exchange properties of the T3 imino proton indicate hydrogen bonding to A6 N7 via a water bridge. The d(5mCCTTTTCC) deoxyoligonucleotide, in which position 6 is occupied by a thymidine instead of an adenine, also forms a symmetric i-motif dimer. However, in this structure the two TTTT loops are located on the same side of the i-motif core and the C.C+ pairs are formed by equivalent cytidines stacked in the order 8*.8/1.1*/7*.7/2.2*. CONCLUSIONS: Oligodeoxynucleotides containing two C-rich repeats can fold and dimerize into an i-motif. The change of folding topology resulting from the substitution of a single nucleoside emphasizes the influence of the loop residues on the i-motif structure formed by two folded strands. PMID- 9331415 TI - N-terminal arm exchange is observed in the 2.15 A crystal structure of oxidized nitrite reductase from Pseudomonas aeruginosa. AB - BACKGROUND: Nitrite reductase from Pseudomonas aeruginosa (NiR-Pa) is a dimer consisting of two identical 60 kDa subunits, each of which contains one c and one d1 heme group. This enzyme, a soluble component of the electron-transfer chain that uses nitrate as a source of energy, can be induced by the addition of nitrate to the bacterial growth medium. NiR-Pa catalyzes the reduction of nitrite (NO2-) to nitric oxide (NO); in vitro, both cytochrome c551 and azurin are efficient electron donors in this reaction. NiR is a key denitrification enzyme, which controls the rate of the production of toxic nitric oxide (NO) and ultimately regulates the release of NO into the atmosphere. RESULTS: The structure of the orthorhombic form (P2(1)2(1)2) of oxidized NiR-Pa was solved at 2.15 A resolution, using molecular replacement with the coordinates of the NiR from Thiosphaera pantotropha (NiR-Tp) as the starting model. Although the d1-heme domains are almost identical in both enzyme structures, the c domain of NiR-Pa is more like the classical class I cytochrome-c fold because it has His51 and Met88 as heme ligands, instead of His17 and His69 present in NiR-Tp. In addition, the methionine-bearing loop, which was displaced by His17 of the NiR-Tp N-terminal segment, is back to normal in our structure. The N-terminal residues (5/6-30) of NiR-Pa and NiR-Tp have little sequence identity. In Nir-Pa, this N-terminal segment of one monomer crosses the dimer interface and wraps itself around the other monomer. Tyr10 of this segment is hydrogen bonded to an hydroxide ion--the sixth ligand of the d1-heme Fe, whereas the equivalent residue in NiR-Tp, Tyr25, is directly bound to the Fe. CONCLUSIONS: Two ligands of hemes c and d1 differ between the two known NiR structures, which accounts for the fact that they have quite different spectroscopic and kinetic features. The unexpected domain crossing by the N-terminal segment of NiR-Pa is comparable to that of 'domain swapping' or 'arm exchange' previously observed in other systems and may explain the observed cooperativity between monomers of dimeric NiR-Pa. In spite of having similar sequence and fold, the different kinetic behaviour and the spectral features of NiR-Pa and NiR-Tp are tuned by the N-terminal stretch of residues. A further example of this may come from another NiR, from Pseudomonas stutzeri, which has an N terminus very different from that of the two above mentioned NiRs. PMID- 9331416 TI - The structure of an essential splicing element: stem loop IIa from yeast U2 snRNA. AB - BACKGROUND: Eukaryotic genes are usually transcribed as precursor mRNAs which are then spliced, removing introns to produce functional mRNAs. Splicing is performed by the spliceosome and provides an important level of post-translational control of gene expression. Stem loop IIa from U2 small nuclear (sn)RNA is required for the efficient association of the U2 small nuclear ribonuclear protein (snRNP) with the nascent spliceosome in yeast. Genetic analysis suggests that stem loop IIa is involved in RNA-protein interactions early in splicing, and it may also interact with other RNA sequences in U2. The sequence of loop IIa is well conserved, consistent with the idea that this loop is important for function. RESULTS: We have solved the structure of U2A, a 20-base analogue of stem loop IIa from Saccharomyces cerevisiae, using NMR and restrained molecular dynamics. In the process, we have demonstrated the efficacy of a new structure calculation protocol, torsion angle molecular dynamics. The structure that has emerged, which is consistent with the in vivo chemical protection data available for stem loop IIa in the context of intact U2 snRNA, contains a sheared GA pair followed by a U turn in the loop. The U-turn conformation, which resembles the U-turns in tRNA anticodon loops, makes this stretch of U2 snRNA an obvious target for interactions with proteins and/or other RNA sequences. CONCLUSIONS: The phenotypes of many stem loop IIa mutants can be rationalized assuming that the U turn conformation in the loop must be preserved for efficient splicing. This observation, combined with the phylogenetic conservation of its sequence, suggests that the conformation of the loop of stem loop IIa is essential for its function in pre-mRNA splicing. PMID- 9331417 TI - New angles on actin dynamics. AB - Actin is now realised to play a dynamic role in muscle contraction and many cellular motility events that occur when the motor domain of myosin uses the energy of ATP hydrolysis to move along the actin filament. Optical and electron microscopic studies have led to seemingly contradictory pictures of actin filament dynamics. PMID- 9331418 TI - The structure of ribosomal protein S7 at 1.9 A resolution reveals a beta-hairpin motif that binds double-stranded nucleic acids. AB - BACKGROUND: Ribosomal protein S7, a crucial RNA-binding component of the ribosome, is one of two proteins that initiates assembly of the 30S ribosomal subunit. It is required for proper folding of a large 3' domain of 16S ribosomal RNA. S7 regulates its own synthesis by binding to its own mRNA. This ability of S7 to bind both messenger and ribosomal RNAs makes determination of its mode of RNA recognition particularly interesting. RESULTS: The crystal structure of S7 from Thermus thermophilus was determined by a two-wavelength anomalous diffraction experiment using the LIII edge of mercury. The S7 structure consists of a bundle of six helices and an extended beta hairpin between helices 3 and 4, with two or more RNA-binding sites on its surface. The hairpin, along with portions of helices 1, 4 and 6, forms a large, positively charged, concave surface that has the appropriate curvature and dimensions to bind double-stranded RNA. A second putative RNA-binding site comprises parts of loop 2 and the helix 4 loop 5 turn. CONCLUSIONS: Structural similarity between S7 and the IHF/HU family of proteins strongly suggests that the beta hairpin of S7 binds to a groove of double-stranded RNA. The beta hairpin of S7 is also similar to those from other nucleic acid binding proteins, such as ribosomal protein L14 and BIV Tat, suggesting that it belongs to an extended family of such motifs, all of which bind to a groove of double-stranded nucleic acid. The residues in S7 loop 2 that belong to the second putative RNA-binding site may have a role analogous to the N terminal residues of IHF/HU which grip an unbent portion of double helix. PMID- 9331419 TI - Crystal structure of the A3 domain of human von Willebrand factor: implications for collagen binding. AB - BACKGROUND: Bleeding from a damaged blood vessel is stopped by the formation of a platelet plug. The multimeric plasma glycoprotein, von Willebrand factor (vWF), plays an essential role in this process by anchoring blood platelets to the damaged vessel wall under conditions of high shear stress. This factor mediates platelet adhesion by binding both to collagen of the damaged blood vessel and to glycoprotein Ib on the platelet membrane. The A3 domain of vWF allows it to bind to collagen types I and III present in the perivascular connective tissue of the damaged vessel wall. To gain insight into the mechanism of collagen binding by vWF, we have determined the crystal structure of the human vWF A3 domain. RESULTS: The crystal structure of the 20 kDa A3 domain of human vWF (residues 920 1111), determined by the method of multiwavelength anomalous dispersion at 1.8 A resolution, exhibits a common dinucleotide-binding fold. The putative collagen binding site of the A3 domain is rather smooth and shows a markedly high concentration of negatively charged residues. This region encompasses a potential metal-binding site containing the motif DXSXS, which is required for ligand interaction in the homologous I-type domains of integrins CR3 and LFA-1. Although vWF A3 has considerable sequence and structural similarity with CR3 and LFA-1 in this region, one loop of A3 adopts a conformation which is incompatible with ion binding. CONCLUSIONS: The structure of the A3 domain suggests that adhesion to collagen is primarily achieved through interactions between negatively charged residues on A3 and positively charged residues on collagen. The absence of a pronounced binding groove precludes a large van der Waals surface interaction between A3 and collagen and is consistent with the low affinity for collagen of a single A3 domain and the requirement for multimeric vWF for tight association with collagen. The absence of bound metal ions upon soaking the crystal in MgCl2 and vWF A3's conformational incompatibility for metal binding is consistent with the absence of a functional role for metal ion binding in A3, which contrasts the metal ion activation required for ligand binding by the homologous integrin I type domains. PMID- 9331421 TI - Helicase structures: a new twist on DNA unwinding. AB - The crystal structures of two members of the SF1 family of helicases, Rep and PcrA, and one member of the SF2 family of helicases, the HCV RNA helicase, have recently been solved. These structures illuminate the roles of the conserved helicase motifs in catalytic function and offer clues as to how these proteins can translocate along DNA. PMID- 9331420 TI - The crystal structure of bovine bile salt activated lipase: insights into the bile salt activation mechanism. AB - BACKGROUND: The intestinally located pancreatic enzyme, bile salt activated lipase (BAL), possesses unique activities for digesting different kinds of lipids. It also differs from other lipases in a requirement of bile salts for activity. A structure-based explanation for these unique properties has not been reached so far due to the absence of a three-dimensional structure. RESULTS: The crystal structures of bovine BAL and its complex with taurocholate have been determined at 2.8 A resolution. The overall structure of BAL belongs to the alpha/beta hydrolase fold family. Two bile salt binding sites were found in each BAL molecule within the BAL-taurocholate complex structure. One of these sites is located close to a hairpin loop near the active site. Upon the binding of taurocholate, this loop becomes less mobile and assumes a different conformation. The other bile salt binding site is located remote from the active site. In both structures, BAL forms similar dimers with the active sites facing each other. CONCLUSIONS: Bile salts activate BAL by binding to a relatively short ten-residue loop near the active site, and stabilize the loop in an open conformation. Presumably, this conformational change leads to the formation of the substrate binding site, as suggested from kinetic data. The BAL dimer observed in the crystal structure may also play a functional role under physiological conditions. PMID- 9331422 TI - Crystal structure of the epsilon subunit of the proton-translocating ATP synthase from Escherichia coli. AB - BACKGROUND: Proton-translocating ATP synthases convert the energy generated from photosynthesis or respiration into ATP. These enzymes, termed F0F1-ATPases, are structurally highly conserved. In Escherichia coli, F0F1-ATPase consists of a membrane portion, F0, made up of three different polypeptides (a, b and c) and an F1 portion comprising five different polypeptides in the stoichiometry alpha 3 beta 3 gamma delta epsilon. The minor subunits gamma, delta and epsilon are required for the coupling of proton translocation with ATP synthesis; the epsilon subunit is in close contact with the alpha, beta, gamma and c subunits. The structure of the epsilon subunit provides clues to its essential role in this complex enzyme. RESULTS: The structure of the E. coli F0F1-ATPase epsilon subunit has been solved at 2.3 A resolution by multiple isomorphous replacement. The structure, comprising residues 2-136 of the polypeptide chain and 14 water molecules, refined to an R value of 0.214 (Rfree = 0.288). The molecule has a novel fold with two domains. The N-terminal domain is a beta sandwich with two five-stranded sheets. The C-terminal domain is formed from two alpha helices arranged in an antiparallel coiled-coil. A series of alanine residues from each helix form the central contacting residues in the helical domain and can be described as an 'alanine zipper'. There is an extensive hydrophobic contact region between the two domains providing a stable interface. The individual domains of the crystal structure closely resemble the structures determined in solution by NMR spectroscopy. CONCLUSIONS: Sequence alignments of a number of epsilon subunits from diverse sources suggest that the C-terminal domain, which is absent in some species, is not essential for function. In the crystal the N terminal domains of two epsilon subunits make a close hydrophobic interaction across a crystallographic twofold axis. This region has previously been proposed as the contact surface between the epsilon and gamma subunits in the complete F1 ATPase complex. In the crystal structure we observe what is apparently a stable interface between the two domains of the epsilon subunit, consistent with the fact that the crystal and solution structures are quite similar despite close crystal packing. This suggests that a gross conformational change in the epsilon subunit, to transmit the effect of proton translocation to the catalytic domain, is unlikely, but cannot be ruled out. PMID- 9331423 TI - Ribosomal protein S7: a new RNA-binding motif with structural similarities to a DNA architectural factor. AB - BACKGROUND: The ribosome is a ribonucleoprotein complex which performs the crucial function of protein biosynthesis. Its role is to decode mRNAs within the cell and to synthesize the corresponding proteins. Ribosomal protein S7 is located at the head of the small (30S) subunit of the ribosome and faces into the decoding centre. S7 is one of the primary 16S rRNA-binding proteins responsible for initiating the assembly of the head of the 30S subunit. In addition, S7 has been shown to be the major protein component to cross-link with tRNA molecules bound at both the aminoacyl-tRNA (A) and peptidyl-tRNA (P) sites of the ribosome. The ribosomal protein S7 clearly plays an important role in ribosome function. It was hoped that an atomic-resolution structure of this protein would aid our understanding of ribosomal mechanisms. RESULTS: The structure of ribosomal protein S7 from Bacillus stearothermophilus has been solved at 2.5 A resolution using multiwavelength anomalous diffraction and selenomethionyl-substituted proteins. The molecule consists of a helical hydrophobic core domain and a beta ribbon arm extending from the hydrophobic core. The helical core domain is composed of a pair of entangled helix-turn-helix motifs; the fold of the core is similar to that of a DNA architectural factor. Highly conserved basic and aromatic residues are clustered on one face of the S7 molecule and create a 16S rRNA contact surface. CONCLUSIONS: The molecular structure of S7, together with the results of previous cross-linking experiments, suggest how this ribosomal protein binds to the 3' major domain of 16S rRNA and mediates the folding of 16S rRNA to create the ribosome decoding centre. PMID- 9331424 TI - 'Flu' and structure-based drug design. AB - The threat of a catastrophic outbreak of influenza is ever present. Vaccines are only partially effective and the two compounds, amantidine and rimantidine, used clinically against influenza A cause side-effects and rapid viral resistance. Recent advances bring hope that specific and potent drugs against influenza may soon be available in the clinic. These compounds were designed to inhibit influenza neuraminidase (NA), one of the viral coat glycoproteins, using the crystal structure of NA which was first published in 1983. In this review, the application of structure-based drug design approaches to the design of anti influenza agents targeted at NA and haemagglutinin (HA), the other viral surface glycoprotein, is discussed. PMID- 9331425 TI - Obituary: Irving Geis, 1908-1997. PMID- 9331427 TI - Role of c-Src tyrosine kinase in EGF-induced mitogenesis. AB - c-Src, the prototype of the cytoplasmic, membrane-associated,non-receptor tyrosine kinases, is a co-transducer of mitogenic signals emanating from a number of tyrosine kinase polypeptide growth factor receptors. Examples of such receptors include those that bind the platelet-derived growth factor (PDGF), colony stimulating factor-1 (CSF-1), and epidermal growth factor (EGF). Investigations into the mechanisms by which c-Src contributes to receptor signaling suggest that interactions between the two proteins are bidirectional, i.e., that c-Src can bind, phosphorylate, and activate the receptor, and vice versa. The consequences of these interactions appear to be enhanced phosphorylation of specific substrates. Delineating which cellular proteins are substrates of which tyrosine kinase and determining the consequences of tyrosine phosphorylation on the function of specific substrates are the goals of current investigations. Utilizing the murine C3H10T fibroblast model, in which a panel of wild type and mutant c-Src/EGF receptor overexpressors has been studied for temporal and spatial second messenger responses to EGF, distinctions between substrates of c-Src and the EGF receptor and the effects of tyrosine phosphorylation on substrate function are beginning to emerge. In the 10T model, preferred substrates of c-Src are almost exclusively comprised of those molecules that associate with the actin cytoskeleton or with focal adhesions, such as cortactin, p190RhoGAP, and p130CAS, while preferred substrates of the EGF receptor include the receptor itself, SHC, phospholipase C-gamma and p62DOK. While the major mitogenic signaling pathway is thought to proceed directly from the receptor (through SHC/GRB2/SOS/Ras/Raf/MEK/MAPkinase/Elk1), more evidence is accumulating to suggest that proteins involved in regulating the actin cytoskeleton (such as c-Src substrates) also participate in mitogenesis, either as unique transducers of growth signals and/or as monitors of anti-apoptotic conditions (substratum attachment). How c-Src may contribute to the EGF mitogenic response through tyrosine phosphorylation of or association with its specific substrates is discussed. Cellular Src (c-Src), prototype for a family of intracellular membrane-associated tyrosine kinases, is required for mitogenesis initiated by multiple growth factor receptors, including the receptors for epidermal growth factor (EGF), platelet-derived growth factor (PDGF), colony stimulating factor-1 (CSF-1), and the basic fibroblast growth factor (bFGF). C Src is also overexpressed and/or activated in many of the same human carcinomas that overexpress members of the EGF receptor (EGFR) family, suggesting that the two types of tyrosine kinases can cooperate during the genesis of human tumors. This review focuses on the role of c-Src in EGF-dependent mitogenesis and tumorigenesis, i.e., on the interactions between c-Src and the receptor and on identification of c-Src substrates, their functions, and the effects of tyrosine phosphorylations on their functions. A synopsis of other mitogenic and signaling systems is also included for comparative purposes. PMID- 9331428 TI - Proceedings of the 3rd European Conference on Vaccinology: Building Life-Long Immunity. Berlin, Germany. PMID- 9331430 TI - The Pathology of Congenital Heart Disease: A Personal Experience with More Than 6300 Congenitally Malformed Hearts PMID- 9331429 TI - REPLY PMID- 9331431 TI - FDA announces tighter regulation of oral levothyroxine products. PMID- 9331432 TI - Bromfenac marketed for short-term pain relief. PMID- 9331433 TI - Proposed regulations would mandate pediatric testing for many drugs. PMID- 9331435 TI - Healthy people 2000. PMID- 9331434 TI - HMG-CoA reductase inhibitors reduce stroke risk, total mortality. PMID- 9331436 TI - Recognizing a nurse as team leader. PMID- 9331437 TI - Direct-to-consumer prescription drug advertising. PMID- 9331438 TI - Intranasal mupirocin for outbreaks of methicillin-resistant Staphylococcus aureus. AB - The pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosage and administration of mupirocin are reviewed. Mupirocin is a naturally occurring antibiotic produced by submerged fermentation of Pseudomonas fluorescens. It inhibits bacterial protein synthesis by binding reversibly and specifically to isoleucyl-tRNA synthetase. Organisms resistant to other antimicrobials are not simultaneously resistant to mupirocin. Mupirocin is highly active against Staphylococcus aureus and other staphylococci and streptococci. When mupirocin ointment is applied topically, local concentrations exceed the inhibitory concentrations for staphylococci and remain detectable for up to 72 hours. Placebo-controlled studies demonstrate the ability of mupirocin to eliminate nasal carriage of S. aureus in health care workers. Observational studies suggest that mupirocin is efficacious in treating methicillin-resistant S. aureus (MRSA) outbreaks. Preliminary studies show that mupirocin might have a role in preventing infections in high-risk patients. Although mupirocin seems to be well tolerated, mild to moderate adverse events have been reported, including respiratory problems and effects confined to the nose--erythema, swelling, burning or stinging, pruritus, and dryness. Mupirocin calcium ointment has FDA approved labeling for the eradication of nasal MRSA colonization in adult patients and health care workers as part of comprehensive infection-control programs to reduce the risk of infection during institutional outbreaks. The recommended dosage is 0.5 g inserted into each nostril twice daily for five days. Intranasal mupirocin ointment appears to be a useful addition to infection control programs designed to reduce the risk of infection among patients during MRSA outbreaks. PMID- 9331439 TI - Compatibility of remifentanil hydrochloride with selected drugs during simulated Y-site administration. AB - The compatibility of remifentanil hydrochloride with 90 other drugs during simulated Y-site administration was studied. Five milliliters of remifentanil 25 and 250 micrograms/mL (as hydrochloride) in 0.9% sodium chloride injection or 5% dextrose injection was combined with 5 mL of each of 90 other drugs in 5% dextrose injection of 0.9% sodium chloride injection. Each combination was prepared in duplicate. The combinations were stored at approximately 23 degrees C under fluorescent light and examined with the unaided eye and in high-intensity monodirectional light during the first 15 minutes after preparation and at one and four hours. The turbidity of each combination was measured as well. Particle sizing and counting were performed for selected combinations. Most of the combinations exhibited no haze, turbidity, or color change throughout the study period. Remifentanil 25 micrograms/mL combined with chlorpromazine hydrochloride showed a small increase in haze within four hours. One of the combinations of remifentanil 250 micrograms/mL with cefoperazone sodium was unacceptably hazy within one hour. The combination of remifentanil 250 micrograms/mL with amphotericin B formed a gross precipitate upon mixing. Remifentanil 25 and 250 microgram/mL (as hydrochloride) in 0.9% sodium chloride injection was compatible for four hours at approximately 23 degrees C with all the drugs studied except chlorpromazine hydrochloride (with remifentanil 25 micrograms/mL), cefoperazone sodium (with remifentanil 250 micrograms/mL), and amphotericin B (with remifentanil 250 micrograms/mL in 5% dextrose injection). PMID- 9331440 TI - Effect of including both physicians and pharmacists in an asthma drug-use review intervention. PMID- 9331441 TI - Effect of body weight on aminoglycoside pharmacokinetics in patients with hypoalbuminemia. PMID- 9331442 TI - Stability of cidofovir in 0.9% sodium chloride injection for five days. PMID- 9331443 TI - Financial risk management of pharmacy benefits. AB - Financial risk management of pharmacy benefits in integrated health systems is explained. A managed care organization should assume financial risk for pharmacy benefits only if it can manage the risk. Horizontally integrated organizations often do not have much control over the management of drug utilization and costs. Vertically integrated organizations have the greatest ability to manage pharmacy financial risk; virtual integration may also be compatible. Contracts can be established in which the provider is incentivized or placed at partial or full risk. The main concerns that health plans have with respect to pharmacy capitation are formulary management and the question of who should receive rebates from manufacturers. The components needed to managed pharmacy financial risk depend on the type of contract negotiated. Health-system pharmacists are uniquely positioned to take advantage of opportunities opening up through pharmacy risk contracting. Functions most organizations must provide when assuming pharmacy financial risk can be divided into internal and external categories. Internally performed functions include formulary management, clinical pharmacy services and utilization management, and utilization reports for physicians. Functions that can be outsourced include claims processing and administration, provider- and customer support services, and rebates. Organizations that integrate the pharmacy benefit across the health care continuum will be more effective in controlling costs and improving outcomes than organizations that handle this benefit as separate from others. Patient care should not focus on payment mechanisms and unit costs but on developing superior processes and systems that improve health care. PMID- 9331444 TI - Thrombolysis in acute ischemic stroke. PMID- 9331445 TI - Lessons learned from projects in disease management in ambulatory care. PMID- 9331446 TI - Home infusion of vasopressin for gastrointestinal bleeding. PMID- 9331448 TI - Our professional responsibility after hospitals change ownership. PMID- 9331447 TI - Home collection and non-blood-based methods of testing for the human immunodeficiency virus. PMID- 9331449 TI - Inconsistencies in labeling of enteral nutrition products. PMID- 9331450 TI - Hormone measures in finger-prick blood spot samples: new field methods for reproductive endocrinology. AB - Comparative endocrine studies have notably advanced understanding of ecological factors that contribute to variation in human reproductive function. Such research has relied on methodological advances that permit hormone determinations in samples that are easily and safely collected, stored, and transported, most recently on measurement of steroids in saliva. This report seeks to further expand the scope of endocrine research by demonstrating the value of blood spot samples collected by finger prick. As a sampling strategy, finger-prick blood spot collection offers the advantages of short collection time, low invasiveness, repeatability, absence of postcollection processing, low biohazard risk, and ease of sample storage and transport. We document good sample stability and present sensitive assay methods for a range of steroids and proteins (FSH, LH, PRL, T, E2, DHEAS, androstenedione, cortisol, SHGB) in blood spots that require sample volumes of 3-12 microliters and display good reliability, specificity, precision, accuracy, and convertibility of results to plasma/serum equivalent concentrations. Laboratory evaluation was augmented by a feasibility study at a remote site in Papua New Guinea that confirmed validity and stability of blood spot collections under field conditions. Research applications of blood spot sampling are illustrated with a series of studies, including cross-sectional surveys for developmental and life span endocrinology, a longitudinal, population based developmental epidemiologic study of puberty, and serial sampling in a dynamic study of neuroendocrine response to suckling. We conclude that the sampling features and wide range of measurable biomolecules of blood spots do constitute a methodological advance for endocrine research. PMID- 9331451 TI - The Brazilian Xavante Indians revisited: new protein genetic studies. AB - A total of 94 individuals from the Xavante village of Rio das Mortes were variously studied in relation to 28 protein genetic systems. No variation was observed for 15 of them, in accordance with previous studies. Of the remaining 13, four (Rh, Duffy, acid phosphatase, and GC) showed significant departures from the averages obtained in 32 other South American Indian populations. If studies performed in the 1960s are considered, there is indication that no significant changes in this village's gene pool has occurred in the last 30 years. Comparison with two other Xavante populations included nine systems with variation, and for three of them (MNSs, Rh, and Duffy) significant differences were found. Genetically the Rio das Mortes are closer to the Sao Marcos than to the Simoes Lopes Xavantes. A dendrogram considering 25 genetic systems and 33 South American Indian populations was constructed. There the Xavante were grouped together, in two neighboring clusters, with three other tribes who speak Ge languages, But these clusters also present populations who speak other languages, and the reproducibility of the tree is low. South American Indians, at least with this set of markers, do not seem to be clearly classified into defined subgroups. PMID- 9331452 TI - Phylogeographic analysis of pigtail macaque populations (Macaca nemestrina) inferred from mitochondrial DNA. AB - Mitochondrial DNA variation was surveyed in nine populations of the pigtail macaque (Macaca nemestrina), covering all three recognized subspecies in Southeast Asia. To do this, a 2,300 base pair fragment spanning the mitochondrial NAD 3 and NAD 4 genes and flanking tRNA subunits leucine and glycine was targeted for amplification and digested with a battery of 16 restriction endonucleases. Out of a total of 107 individuals, 32 unique haplotypes could be distinguished. Parsimony and neighbor-joining analyses grouped the haplotypes into five strongly supported assemblages representing China/Thailand, Malaysia, Sumatra, Borneo, and Siberut. These results indicate that the mainland and island mtDNA haplotypes are strictly and uniquely limited to the geographic ranges of the recognized morphological subspecies. Cladistic and neighbor-joining analyses indicate that inferred phylogenies of mtDNA haplotypes are congruent with subspecies designations. Furthermore, in support of morphological studies, results indicate that the Mentawai macaque is most likely not a distinct species but a subspecies of M. nemestrina. PMID- 9331453 TI - Primate natal coats: a preliminary analysis of distribution and function. AB - Pelage coloration of infants was compiled for 138 species of primates. Three functional hypotheses--alloparental, infant defense, and paternity cloak--for primate natal coats are tested. Neonatal pelage contrasted with adult pelage in over half of the species examined. Subtle or inconspicuous contrast was more common than flamboyant contrast. Natal coats began to change at 5.7 weeks and disappeared by 18.0 weeks postpartum on average. The first body part to lose natal coloration was the head and/or dorsum in the majority of species. Functional analyses provided no support for the only published hypothesis- alloparental--while providing partial support for two new hypotheses--infant defense and paternity cloak. A significant association between testes weight and natal coat contrast supports a link between mating system and infant contrast. This is discussed in terms of infanticide avoidance. Natal coats are proposed to be categorically differentiated into inconspicuous and flamboyant types, not differentiated by a continuous gradation, such as color. Subspecific differentiation and patterns of shared ancestry are assessed. PMID- 9331455 TI - Relationship of enamel hypoplasia to the pattern of tooth crown growth: a discussion. AB - The defects of enamel hypoplasia can be related to the layered structure of enamel which represents the sequence of development in tooth crowns. From such studies, it is possible to see that furrow-type enamel defects (the most common form of hypoplasia seen with the naked eye) are just the most prominent expression of a continuum which extends ever smaller, down to a microscopic disturbance to a single layer in the crown formation sequence. Furthermore, the progressive decrease in spacing between development layers which occurs down the crown sides, from occlusal to cervical, affects both the prominence and apparent width of the defects. This makes it difficult to use measurements as a means of estimating the duration of the disturbance causing a particular defect. The difficulty is even greater for the less common pitted or exposed-plane-type defects, for which the apparent width bears very little relationship with the duration of the growth disturbance. The defects of enamel hypoplasia can therefore be understood clearly only when examined under the microscope in relation to the structures which mark the development sequence of the tooth crown. PMID- 9331454 TI - Dental caries in nineteenth century upper Canada. AB - This study examines the presence of dental caries in a large sample of adult skeletons from the 19th century cemetery of St. Thomas' Anglican Church in Belleville, Ontario. The cemetery was used from 1821 to 1874. Caries prevalence and frequencies of diseased and missing teeth were calculated both by observing summary statistics of individual rates and by the total sample of teeth. Postmortem tooth loss is low in this sample and antemortem tooth loss is highest in first mandibular molars, all other molars and then premolars. Age at death, but not sex, was found to be significantly related to the overall Caries Rate while both age and sex were significantly associated with the Diseased-Missing Index. The increase in diseased and missing teeth in older individuals is expected while the sex difference is not explained by simple dietary factors. When compared to reports on British and American samples, caries and antemortem tooth loss in the St. Thomas' sample is most similar to a pre-1850 British group and higher than American samples. Although there is undoubtedly a complex of factors contributing to caries prevalence in this sample, more data are required from large historic samples, particularly from the American northeast and late 19th century Britain, to have a clearer understanding of the influence of diet, cultural, and environmental factors affecting caries rates in historic populations. PMID- 9331456 TI - Functional osteology of the primate carpus with special reference to strepsirhini. AB - Preuschoft et al. ([1993] in H. Preuschoft and D. Chivers (eds): Hands of Primates. New York: Springer-Verlag, pp. 245-256) used a theoretical biomechanical analysis to generate several predictions relating subordinal differences in primate hand proportions to differences in carpal morphology. This study tests these predictions using quantitative analyses of carpal morphology between extant haplorhine and strepsirhine primates. Results show that living strepsirhines have a significantly larger hamate hamulus than do haplorhines, supporting a Preuschoft et al.'s (1993) predictions. Extant strepsirhines also have a significantly shorter pisiform body than do haplorhines and arboreal nonprimate eutherians and a larger scaphoid tubercle than new and Old World monkeys. These results contrast markedly with those expected under Preuschoft et al.'s (1993) model. Furthermore, strepsirhines and haplorhines do not differ significantly in the relative size of their radiocarpal articulations. These morphometric observations do not match the predicted morphological patterns because the kinematic assumptions upon which the biomechanical models are based are incorrect. Living strepsirhines appear to be derived in having very deep radial and ulnar margins of the carpal tunnel for well-developed extrinsic digital flexors. Moreover, tooth-combed prosimians differ from most haplorhines, early Tertiary adapiforms, and arboreal nonprimate eutherians in having a relatively short pisiform body, which gives the flexor carpi ulnaris less power to flex the wrist from extended (= dorsiflexed) positions. These structural observations suggest that powerful manual grasping and an emphasis on leaping and climbing, rather than palmigrade quadrupedal walking and running, are morphotypic for extant Strepsirhini. PMID- 9331457 TI - Cortical bone distribution in the femoral neck of hominoids: implications for the locomotion of Australopithecus afarensis. AB - Contiguous high resolution computed tomography images were obtained at a 1.5 mm slice thickness perpendicular to the neck axis from the base of the femoral head to the trochanteric line in a sample of 10 specimens each of Homo sapiens, Pan troglodytes, and Gorilla gorilla, plus five specimens of Pan paniscus. Superior, inferior, anterior, and posterior cortical thicknesses were automatically measured directly from these digital images. Throughout the femoral neck H. sapiens displays thin superior cortical bone and inferior cortical bone that thickens distally. In marked contrast, cortical bone in the femoral neck of African apes is more uniformly thick in all directions, with even greater thickening of the superior cortical bone distally. Because the femoral neck acts as a cantilevered beam, its anchorage at the neck-shaft junction is subjected to the highest bending stresses and is the most biomechanically relevant region to inspect for response to strain. As evinced by A.L. 128-1, A.L. 211-1 and MAK-VP 1/1, Australopithecus afarensis is indistinguishable from H. sapiens, but markedly different from African apes in cortical bone distribution at the femoral neck-shaft junction. Cortical distribution in the African ape indicates much greater variation in loading conditions consistent with their more varied locomotor repertoire. Cortical distribution in hominids is a response to the more stereotypic loading pattern imposed by habitual bipedality, and thin superior cortex in A. afarensis confirms the absence of a significant arboreal component in its locomotor repertoire. PMID- 9331458 TI - Patterns of femoral bone remodeling dynamics in a Medieval Nubian population. AB - The relationship between age, sex and histomorphometry in femoral cortical bone was examined in a skeletal population of late Medieval antiquity (AD 1250-1450) from Kulubnarti, in Sudanese Nubia. These skeletal remains are naturally mummified and in an excellent state of preservation. The study sample consisted of femoral cross sections from 24 females and 19 males ranging in age from 20 to 50+ years. Femoral cross sections were examined using an image analysis system. Numbers of secondary osteons and osteon fragments were counted, osteon area and Haversian canal area were measured, and several variables were calculated to assess differences between sexes and among age groups in bone remodeling variables. The results indicate significant differences between the sexes in osteon number and size. Males had significantly more intact osteons than females, whereas females had significantly larger osteons than males. Haversian canal dimensions were not statistically significant between the sexes. Sex differences in activity patterns in which males were involved in more physically strenuous tasks may have contributed to differences in remodeling variables. Interpopulational comparisons suggest that mechanical strain affects the microstructural features examined in this study. In particular, small Haversian canals in some archaeological skeletal populations are associated with higher bone volume, which may result from high levels of mechanical strain. PMID- 9331469 TI - A computational model of the response of honey bee antennal lobe circuitry to odor mixtures: overshadowing, blocking and unblocking can arise from lateral inhibition. AB - Recent studies of learning about elements of odorant mixtures in honey bees identified several types of interactions between mixture components, such as overshadowing and blocking. The latter phenomenon in particular indicates at least a limited ability of subjects to identify the most salient element of a binary mixture. Here we show that the circuitry in the antennal lobes, the first neuropil in which synaptic interaction affects sensory processing, could give rise to both effects given the incorporation of modifiable synapses onto inhibitory circuitry. The neural model of the antennal lobe that we present incorporates identified cell types and includes a biologically realistic modulatory neuron with which modifiable Hebb-like synaptic interactions take place. A learning rule that incorporates modifiable connections from output (projection) neurons onto the modulatory neuron is sufficient to account for behavioral results on generalization and overshadowing. A second type of excitatory connection from the modulatory neuron onto local inhibitory interneurons is necessary to reproduce behavioral results from blocking and unblocking. We suggest that the neural representations of odor mixtures in the antennal lobe can be modified by previous exposure to one of the mixture components. These results provide testable hypotheses that will guide future behavioral and physiological analyses. PMID- 9331470 TI - Forced-choice discrimination of equimolar NaCl and LiCl solutions in rats: effects of ablating the chemosensitive area postrema on acquisition and retention. AB - The area postrema (AP), a chemosensitive organ located in the fourth ventricle, has been shown to mediate the formation of a lithium-induced conditioned taste avoidance (CTA) in rats. The present experiments examined the role of the AP in the discrimination between two equimolar solutions of sodium chloride (NaCl) and lithium chloride (LiCl). In the first experiment adult male rats were trained to discriminate between equimolar (0.12 M) solutions of NaCl and LiCl in a forced choice procedure over a 10-day acquisition phase. Subsequently half of the rats (n = 7) received AP lesions (APX) and the other half (n = 7) were given sham lesions (SHAM). In the retention phase all animals were again exposed to the same salt solutions over a 10-day period. Good discrimination (P < 0.001) between the two salt solutions was demonstrated by the end of the acquisition phase and both the APX and SHAM groups exhibited robust retention (P < 0.01) of this discrimination in the second phase. However, when only a LiCl solution was available the APX group ingested significantly more (P < 0.01) than the SHAM rats. No significant group difference emerged when only NaCl was available. In the second experiment rats received ablations of AP or sham lesions and were then trained to discriminate between 0.12 M NaCl and LiCl solutions in a forced-choice procedure over a 10-day period. Both groups exhibited a clear discrimination (P < 0.01) between the two solutions by the end of the acquisition phase. APX rats ingested significantly more LiCl (P < 0.01) than did the SHAM group when this was the only type of fluid available. Again, no such difference was evident when only NaCl was available. These experiments demonstrate that the AP is not necessary for either the acquisition or retention of a discrimination between equimolar solutions of NaCl and LiCl in a forced-choice procedure and that this discrimination is not mediated by a conditioned taste aversion to the LiCl solution. PMID- 9331471 TI - Patterns of cognitive decline in aged rhesus monkeys. AB - Although cognitive decline has been well established as a consequence of aging in non-human primate models, the prevalence or frequency of impairment for specific age ranges has not been described. The first aim of this study was to estimate prevalence of cognitive impairment on each of the six tests of cognitive performance by comparing the performance of early-aged (19-23 years old), advanced-aged (24-28 years old), and oldest-aged (29+ years old) monkeys to that of young adults (< 15 years old). The second aim was to derive a single overall measure of cognitive performance to help classify behavioral function in our aged monkeys. Accordingly, we obtained performance measures for these age groups on six behavioral measures: (1) acquisition of the delayed non-matching-to-sample task (DNMS); (2) performance of the DNMS with a delay of 120 sec; (3) the spatial condition of the delayed recognition span test (DRST); (4) the color condition of the DRST; (5) spatial reversal learning; and (6) object reversal learning. Early aged monkeys displayed prevalence rates of impairment significantly greater than zero on all tasks except the DRST-color. The highest prevalence of impairment was observed in this age group in a task measuring spatial memory (DRST). Significant trends toward progressively higher impairment rates in advanced-aged and oldest aged monkeys were observed for DNMS-acquisition, DRST-color and spatial reversal learning tasks. A linear transformation of standardized scores on the six cognitive tests was derived by means of principal components analysis (PCA). The first PCA (PCA1) included data from 30 monkeys with available data on all six measures, and yielded a composite measure which declined linearly with increasing age (r = -0.74). A second PCA (PCA2) was performed on data from 53 monkeys for which three test scores (DNMS-acquisition, DNMS-120s delay, and DRST-spatial condition) were available. The composite score derived from this analysis was highly correlated (r = 0.93) with the composite score from PCA1, suggesting that a score based on only three tests may provide an adequate classification of global cognitive ability. PMID- 9331473 TI - A neural mechanism of the generation of meaning in cognitive processes. AB - A neural mechanism for control of computational dynamics underlying the generation of meaning in cognitive processes is demonstrated. Meaning derives from recognition of connections between items in related conceptual classes of the neural representation in the brain. It is generated from a stimulated item in one conceptual class by an associative process in which linked items in related conceptual classes are activated in parallel. The complexity of the dynamics of this process varies between exploratory and direct retrieval modes. It is shown that the dynamics mode of the generation of meaning can be controlled by the neuronal adaptivity, i.e., the coupling strength between neuronal activity and excitability. Neuronal adaptivity in turn is controlled by neuromodulators in the brain. An autonomous regulation of the dynamics can be accomplished by an activity-dependent release of neuromodulators. The generation of a sequence of bifurcations from an initial exploratory phase to a final retrieval of appropriate item in each conceptual class is demonstrated. The time required for retrieval is shown also to depend on synaptic coupling strengths. The involvement of neuromodulatory systems in cognitive processes has long been observed but the underlying mechanisms not known. The present model describes a mechanism based on the primary effect observed of neuromodulators, viz. that on neuronal adaptivity, and is shown to be consistent with many neuroanatomical, neurophysiological and behavioural observations. PMID- 9331472 TI - Eye-position effects in directional hearing. AB - The influence of gaze direction on azimuthal sound localization was investigated by presenting free-field acoustical stimuli in combination with a visual fixation task. In Experiment 1, a two-alternative forced-choice method was employed. While fixating visual targets, subjects judged whether noise bursts, presented from various directions, were perceived as being on the left or right of either a visual reference indicating straight ahead or the subjective straight-ahead direction. The psychometric functions measured with the first task shifted consistently opposite to the direction of eccentric gaze, i.e., the location of the auditory stimulus was perceived as shifted toward the direction of gaze. The mean magnitude of the shift was 4.7 degrees over a range of fixation angles up to 45 degrees on either side. Without an external reference indicating straight ahead, shifts of sound localization were inconsistent, either opposite or toward the direction of fixation in individual subjects. In Experiment 2, subjects orientated their head toward sound stimuli while fixating visual targets in various directions. As in Experiment 1, head position as a measure of sound localization shifted significantly toward the direction of eccentric gaze when a visual reference of the head median plane was present, and the results were inconsistent across subjects when it was absent. The results indicate a significant effect of gaze direction on the spatial agreement of auditory and visual perception which may be based on the superposition of distinct auditory and visual eye-position effects. The effect is in agreement with previous neurophysiological results that have suggested an incomplete neural transformation of auditory spatial coordinates from a craniocentric into an oculocentric frame of reference. PMID- 9331474 TI - Effect of ischemic cerebral volume changes on behavior. AB - Ischemia causes long-term effects on brain volume and neurologic function but the relationship between the two is poorly characterized. We studied the relationships between brain volume and three measures of rodent behavior after cerebral ischemia was induced by injecting several thousand microspheres into the internal carotid arteries of rats. Forty eight hours later, each subject was rated using a global neurologic rating scale. Several weeks later, the subjects were tested for open field activity and visual spatial learning. Post-mortem we measured the volume of the cerebral hemispheres and estimated the volume densities of cortex, white matter, hippocampus, basal ganglia, thalamus, ventricle, and visible infarction. Ischemia caused significant impairment, as measured by the global rating scale; the probability of an abnormal rating was correlated with the number of microspheres trapped in the brains. Visual spatial learning was significantly impaired by ischemia, but this deficit was independent of the count of microspheres, whether the subject was abnormal at 48 h, and whether the left or right hemisphere was embolized. Cerebral hemisphere volume was reduced from 430 mm3 to 376 mm3 (P < 0.05). The cortex was reduced from 22 to 19% of cerebrum (P < 0.05) and the white matter compartment was reduced to similar degree. The lesion volume was 6% of cerebrum, comparable to that seen with other ischemia methods. The global outcome rating was significantly related to total cerebral volume, but not to volume changes in any single compartment. On the other hand, visual spatial learning was significantly influenced by volume changes in the cortex and white matter, but not by the topography of the visible infarctions. Open field activity was not altered by infarction. Our data suggests that the total volume of brain tissue lost to infarction may partially determine global neurological rating independently of the topography of the volume loss. Integrative functions such as learning may depend more on the integrity of specific compartments and less on the total volume of intact brain. The volume of visible cystic infarction was not related to long term behavioral outcome. These results should be confirmed using another method of inducing ischemia. PMID- 9331475 TI - Electrophysiological correlates of the limbic-motor interactions in various behavioral states in rats. AB - Depth electroencephalographic (EEG) activity was recorded from basolateral amygdala (BLA), ventral subiculum (VSB), n. accumbens (ACC) and subpallidal area (SPL) in freely moving rats, during locomotor tasks with various types of reinforcement in order to compare the strength of limbic-motor interactions in selected behavioral situations. For all EEG signals multichannel coherences (ordinary, multiple and partial) were calculated using autoregression model. Partial coherences indicate the level of synchronization between two signals, thus they were assumed to indicate the strength of direct connection between the structures from which these signals have been recorded. The partial coherences were calculated for six selected frequency bands and the strength of connections within the BLA-VSB-ACC-SPL circuit was estimated for two different behavioral situations and compared. It was found that the strength of connections is sensitive to changes in both motor and emotional aspects of behavioral situation: the strength of BLA-VSB, VSB-ACC, and ACC-SPL depended on motor demands of behavioral task; these of BLA-VSB increased in the highest frequency bands in all emotionally engaging situations when compared with well trained locomotive; the strength of ACC-SPL increased in situations when automatic stereotyped motor behavior was induced by biologically important stimuli, while it decreased or did not change in the motor tasks demanding more precise and quickly adjustable movements. The results are discussed according to the motor-limbic integration model of proposed by Mogenson and show the dynamics of its connections in relation to the motivational-emotional context of the task. PMID- 9331476 TI - Monkeys can associate visual stimuli with reward delayed by 1 s even after perirhinal cortex ablation, uncinate fascicle section or amygdalectomy. AB - In the present experiment monkeys learned concurrent associations of two dimensional objects (presented on a computer screen) with delayed reward. Hypothetical mechanisms of associative memory, such as long-term potentiation (LTP), required coincidental activation of two population of neurons: one representing the object and the other signalling the reward. In monkeys neurons in area TE of temporal cortex show object-specific activity during object presentation but only fraction of those neurons remain active after stimulus offset. In a delayed reward condition the majority of object-specific neurons in TE cease firing before reward is given and can be detected. In the present study the rate of learning with 1000 ms delay of reward was no slower than learning with immediate reward. This indicates that information about the object is somehow retained across the delay, possibly somewhere outside TE. In the present study we tested that assumption. Area TE projects to the perirhinal cortex and, via uncinate fascicle, to the prefrontal cortex. In our hands, ablations of perirhinal cortex or disconnection of prefrontal cortex from TE (by transection of uncinate fascicle) did not impair learning with delayed reward. Ablation of amygdala, a structure involved in reward-learning, slowed down learning equally with and without delay. We conclude that retaining information about the visually perceived objects across a delay does not exclusively depend upon integrity of perirhinal cortex, or uncinate fascicle, or amygdala. Parallel involvement of those structures remains a possibility and establishment of the role of residual activity of TE neurons requires further neurophysiological investigation. PMID- 9331477 TI - Effects of 5-HT2 receptors blockade on fear-induced analgesia elicited by electrical stimulation of the deep layers of the superior colliculus and dorsal periaqueductal gray. AB - The deep layers of the superior colliculus (DLSC) and the dorsal periaqueductal gray matter (DPAG) have been implicated in the control of defensive-like behaviors. Electrical and chemical stimulation of these structures elicits fear and escape behaviour, expressed by immobility (freezing) and wild running, followed by jumps and rapid rotations. There is evidence that the neural substrates responsible for defensive behavior in this level of the midbrain tectum (MT) may also be responsible for fear-induced analgesia. This study was aimed at examining the characteristics of the analgesia that follows the defense oriented reactions induced by electrical midbrain tectum stimulation at freezing and escape thresholds. The animals were submitted to the tail-flick test, following the induction of the defense behavioral responses. The obtained results show that the antinociception that follows the freezing and escape behaviors were not antagonized by MT microinjections of the opioid antagonist naltrexone. These results emphasize previous data showing the non-opioid nature of this analgesia. On the other hand, the fear-induced analgesia was inhibited by microinjections of the serotonergic blockers, methysergide and ketanserin in the MT. Since methysergide is a non-specific antagonist of 5-HT receptors and ketanserin acts with a high degree of specificity at 5-HT2 receptors the present results suggest that activation of 5-HT2 receptors may be implicated in the antinociception induced by midbrain tectum stimulation. PMID- 9331478 TI - Role of 5-HT1B, 5-HT2A and 5-HT2C receptors in learning. AB - The effects of post-training (i.p.) injection of TFMPP, mCPP, DOI or 1-NP in the autoshaping learning task was explored. Furthermore, the post-training effects of these agonists after treatment with the antagonists (+/-)-pindolol, (+/-) propranolol, NAN-190, ketanserin, ritanserin, mesulergine, MDL-72222 or p chloroamphetamine (5-HT depleter) were studied. Rats were individually trained with a lever-press response (conditioned response; CR) on the autoshaping task and tested 24 h later. The results showed that the injection of TFMPP (1-10 mg/kg), mCPP (1-10 mg/kg), 1-NP (0.1-1.0 mg/kg) or mesulergine (0.4 mg/kg) decreased the rate of CR, while DOI (0.01-0.1 mg/kg) and ritanserin (0.5 mg/kg) and ketanserin (0.001-0.1 mg/kg) increased it. However, the effect induced by TFMPP was reversed by (+/-)-pindolol, ketanserin, ritanserin and PCA; the mCPP induced effect was antagonized by (+/-)-propranolol, ketanserin, ritanserin and MDL-72222; and the effect produced by 1-NP was reversed by ketanserin, ritanserin and PCA. In addition, the increment in CR provoked by DOI was enhanced by ketanserin, and reversed by ritanserin, mesulergine and PCA. These findings suggest that TFMPP, 1-NP and DOI exerted their effects via stimulation of presynaptic 5-HT receptors. The effects of mCPP most probably reflect activation of postsynaptic receptors. The present data suggest that both 5-HT1B and 5-HT2A 2C receptors play a significant role in the consolidation of learning. PMID- 9331479 TI - The cost of delaying rewarding brain stimulation. AB - Six rats trained to press a lever to obtain rewarding electrical stimulation of the brain through chronically implanted, lateral hypothalamic electrodes were used to estimate the rate at which short delays between the response and the reward degraded the rewarding effect of the stimulation. Frequency thresholds rose steadily with delays through to 2.2 s at a rate of 10% per second. PMID- 9331480 TI - The effects of frontal cortical lesions on remembering depend on the procedural demands of tasks performed in the radial arm maze. AB - We trained 24 rats to perform an eight-arm radial maze task and then assigned them with a matching procedure to one of three treatments: sham surgery or lesions of the projection areas of the mediodorsal thalamic nucleus (MDn) in the medial wall (MW) or in both the MW and rhinal sulcal (RS) areas of frontal cortex. After recovery we trained the rats to perform six tasks, beginning with the standard eight-arm task, followed by two versions of a four forced choice procedure, and then three versions of a two-choice delayed-nonmatching-to-sample (DNMTS) task. The two lesion groups performed comparably on all tasks, showing that impairments were not exacerbated by extension of the MW lesion to include all cortical areas innervated by MDn. As in previous studies, frontal animals performed the radial maze task poorly immediately after surgery but improved with subsequent training. Controlling the order of the arm entries by opening the first four gates in a random sequence had little effect on performance, although frontal animals were impaired when lengthy delays (5 or 15 min) were imposed after the last of the four forced entries. Frontal animals were not impaired on two-choice DNMTS when the arms used for training were selected at random from the eight alternatives on a trial by trial basis, even when visual cues were eliminated by darkening the room and covering the maze. Frontal animals were significantly impaired when the selection of sample and choice arms was limited to the same two alternatives on every trial. This finding may explain the reported sensitivity of DNMTS to the effects of frontal lesions when training is carried out in operant chambers. PMID- 9331481 TI - Scheduled activity reorganizes circadian phase of Syrian hamsters under full and skeleton photoperiods. AB - Circadian rhythms can be shifted or entrained by light and by arousing nonphotic stimuli. Interactions between photic and nonphotic stimuli were examined by subjecting hamsters to a daily 3 h bout of induced running under full (FPP) or skeleton (two daily light pulses; SPP) photoperiods. Activity scheduled in mid day of a FPP induced large phase delays (260 +/- 63 min) in hamsters that ran more than 4000 rev/3 h. Split rhythms were not evident in constant dark (DD) tests. Activity scheduled in mid-subjective day of a SPP induced 180 degrees inversions of circadian phase, apparently achieved by oscillator splitting in some cases. Activity scheduled late-day and early-night induced a mix of phase delays, advances and no responses. Activity scheduled at two phase ranges late in the night had no effect, but scheduled 1 h later (beginning the last hour of darkness) induced large phase delays (238 +/- 30 min). There was no evidence of oscillator splitting during DD tests, but free-running period was significantly longer in groups that showed large phase delays. Induced running schedules have powerful effects on the phase of photic entrainment and can alter intrinsic pacemaker properties, including internal oscillator coupling and period. PMID- 9331482 TI - Lesions of the basolateral amygdala abolish the ability of drug associated cues to reinstate responding during withdrawal from self-administered cocaine. AB - This study investigated the ability of bilateral excitotoxic lesions of the basolateral amygdala (BLA) to disrupt cocaine self-administration, responding during extinction sessions, and stimulus cued recovery of extinguished responding in rats. BLA and sham lesions following 7 days of 3 h limited access cocaine self administration sessions (0.33 mg/infusion) on a fixed ratio 1 (FR1) schedule of reinforcement failed to alter cocaine intake and responding on 7 subsequent days of self-administration. This lack of effect suggests that the BLA is not paramount for the maintenance of cocaine's reinforcing effects. In contrast, BLA lesions, but not sham lesions, following 7 to 14 days of cocaine self administration attenuated responding on a lever associated with cocaine infusions on the first day of extinction sessions and blocked the ability of drug associated stimuli to reinstate extinguished responding following 20 daily extinction sessions. However, lesions of the BLA did not attenuate stimulus cued recovery of responding following 43 days of withdrawal. These results are consistent with the hypothesis that the BLA is important for the conditioned incentive properties of reinforcers, but not primary reinforcement itself. PMID- 9331483 TI - Effects of arginine8-vasopressin administered at different times in the learning of an appetitive visual discriminative task in mice. AB - A visual discrimination task was used to investigate the effect of the intra hippocampal injection of arginine8-vasopressin (AVP) in male Balb/c mice at different stages of the learning processes. The peptide was bilaterally microinjected at a dose of 25 pg per animal, i.e. 833 pg/kg, into the ventral hippocampus, in a volume of 0.3 microliter 10 min before either the first or the second learning session, or immediately after the first or second learning session. Following pre-session administration of AVP, no effect of the peptide was observed on the session prior to which it was administered. On the other hand, 48 h after the pre-first session treatment, it seems that AVP animals had trouble learning the task. Following post-session injection of AVP, no effect was observed when the treatment was given after the first learning session and a tendency to improve performance was noted when the treatment was given after the second learning session. Thus, whatever time AVP was injected during learning, little or no effect was observed. These results and previous work on the same behavioral task showing a clear enhancing effect of the peptide on retrieval processes, suggest that prior experience or mnemonic context before AVP treatment is as important a factor in understanding the effects of AVP on memory processes as the administration route or the doses used. PMID- 9331484 TI - Unilateral striatal lesions impair response execution on a lateralised choice reaction time task. AB - A novel lateralised reaction time task is described and used to evaluate the effects of D-amphetamine injections and unilateral dorso-striatal lesions in rats. The task involves a two-lever Skinner box adaptation of the nine-hole box visual choice reaction time task first developed by Carli et al. D-Amphetamine had a dose dependent effect on nearly all aspects of task performance. Low and the intermediate doses of D-amphetamine speeded reaction time and movement time, and abolished the delay-dependent pattern or responding in the task. The highest dose of amphetamine disrupted the animals' ability to perform reliably, the task contingencies. Unilateral lesions in the dorsal neostriatum resulted in an increase of error trials, produced a bias to respond towards the ipsilateral side, and decreased the accuracy of responding to contralateral stimuli. The overall mean reaction time to contralateral stimuli was not influenced by the lesions, but the movement time was increased selectively when responding to contralateral stimuli. The data suggest that striatal activation by amphetamine increases motor readiness, which can enhance reaction time performance at the cost of increased errors due to anticipation of cue presentation, in particular at long holding delays. Conversely, striatal lesions induce lateralised defects in executive, rather than sensory, processes, and impair the animals' ability to execute movement towards the contralateral side. PMID- 9331486 TI - Dissociation of spatial reference memory, spatial working memory, and hippocampal mossy fiber distribution in two rat strains differing in emotionality. AB - Rats of the inbred strains DA/Han and BDE/Han were compared on two complex spatial learning tasks, a spatial reference memory task in a 16-unit multiple T maze and a spatial working memory task in an eight-arm radial-maze. In addition, sizes of hippocampal mossy fiber terminal fields were measured. BDE rats showed marked superiority in multiple T-maze learning whereas DA rats outperformed BDE rats on the radial-maze task. DA rats had significantly larger intra- and infrapyramidal mossy fiber terminal fields (IIP-MF). This is consistent with findings from other studies suggesting that large IIP-MF are related to excellent spatial radial-maze learning, but it also indicates that size of IIP-MF is correlated with processing of a specific type of spatial information rather than with overall spatial abilities. BDE rats had more extended suprapyramidal mossy fiber projections (SP-MF) and a larger hilus. Rats of both strains differed in exploratory behaviour and emotionality: DA rats revealed little freezing and had a high rearing activity, whereas BDE rats showed frequent freezing and reared rarely. Results suggest that IIP-MF are involved with flexible expression of memory, updating environmental information and parallel processing whereas SP-MF might be linked to processing of familiar information. Presumably, emotional factors contribute to performance differences. PMID- 9331485 TI - Differential contributions of the two visual pathways to functional lateralization in chicks. AB - The contribution of the two visual pathways to lateralization of visual behaviour in chicks was assessed using unilateral injections of 0.5 microliters of 100 mM monosodium glutamate into localized regions of the forebrain. Chicks treated with glutamate in the left visual hyperstriatum made more errors in a visual discrimination task (pebble-floor test) than did chicks treated in the right visual hyperstriatum. Glutamate injection into the left visual hyperstriatum also elevated attack and copulation scores, but this did not occur following injection of the right visual hyperstriatum. The performance of chicks treated in the right visual hyperstriatum did not differ from that of sham-operated controls. Thus, only the left visual hyperstriatum is involved in the control of these three visually guided behaviours. By contrast, glutamate injections of the left ectostriatum affected only the attack behavior and not performance in the pebble floor test or copulation responses. Glutamate treatment of the right ectostriatum had no affect on any of the behaviours tested and this was also the case for glutamate treatment of both the left and right neostriata. Although injecting glutamate in a larger volume that allows glutamate to spread over a wide area of the left hemisphere is known to retard auditory habituation, localized injection of glutamate in the areas chosen for this study had no effect on auditory habituation. The results suggest that the tectofugal and thalamofugal pathways have different roles in the lateralization of visual functions. The forebrain region which receives input from the thalamofugal visual system has a lateralized role in categorising pebbles as different from food grains, and also a role in controlling attack and copulation responses. The forebrain region which receives input from the tectofugal visual system is involved in the control of attack responses only. PMID- 9331487 TI - Vasopressin and memory. I. The vasopressin analogue AVP4-9 enhances working memory as well as reference memory in the radial arm maze. AB - The present study examined the effects of vasopressin on memory. Healthy rats were injected with the arginine vasopressin fragment AVP4-9 and the AVP antagonist [beta-mercapto-beta,beta-cyclopentamethylenepropionyl1,O-Et-Pyr 2,Val4,Arg8] and were tested in an eight-arm radial maze for 60 sessions. All injections were given s.c. 30 min prior to testing. AVP4-9 enhanced radial arm maze performance. AVP4-9 treated animals showed enhancement in performance as well as increases in the rate of learning, indicating that they learned the task faster. Furthermore, the overall memory enhancement was due to improved working memory as well as to improved reference memory. These results cannot be explained in terms of changes in locomotor activity because an open field test revealed no differences between groups for both of these compounds. The AVP antagonist did not impair performance in the radial maze. It is concluded that AVP4-9 has a more general effect on memory, one that is not limited to a specific type of memory. PMID- 9331488 TI - Vasopressin and memory. II. Lesions to the hippocampus block the memory enhancing effects of AVP4-9 in the radial maze. AB - Previous work has shown that the arginine vasopressin fragment AVP4-9 enhanced overall performance in the radial arm maze. This enhancement effect was due to improved working memory as well as to improved reference memory. The present study uses a behavioral approach to investigate whether these effects of vasopressin on memory are mediated by the hippocampus. Animals received either NMDA lesions to the hippocampus or sham operations and were then treated with AVP4-9 or saline. The testing was performed in an eight-arm radial maze for 60 sessions, and all injections were given s.c. 30 min prior to testing. The hippocampal lesioned animals showed a marked deficit in working memory that was not ameliorated by AVP4-9; however, the improvement in reference memory produced by the compound was as large as in healthy animals. Since the AVP treatment did not differentiate between the two hippocampal groups, the memory enhancing effects of AVP4-9 were blocked by hippocampal lesions. It is concluded that vasopressin has a more general effect on memory and that its site of action includes but is not limited to the hippocampus. PMID- 9331489 TI - New and potent inhibitors of nitric oxide synthase reduce motor activity in mice. AB - Potent inhibitors of nitric oxide synthase (NOS), 3-bromo-7-nitro indazole, 1-(2 trifluoromethylphenyl)imidazole, S-methyl-L-thiocitrulline and 7-nitro indazole, reduced locomotion in mice. These results suggest that activity of NOS and corresponding NO release are of importance for normal locomotion. PMID- 9331490 TI - Shift in the performance of 24-month-old Wistar rats in the Morris water escape task: a comparison across 36 experiments. AB - Spatial discrimination learning in aged rats serves as an animal model of cognitive aging. We assessed the replicability of spatial discrimination performance in the standard Morris water escape task. To this end the learning curves and the performance in a probe trial of 24-month-old outbred Wistar (HsdWin:Wu) control rats from 36 experiments were compared. These experiments had been performed at our laboratory under strictly controlled conditions over a period of 71 weeks. There was a very high variability in the learning curves between experiments. The initial performance level, i.e. the performance during the first session, did not change systematically across the 36 experiments. In contrast, the final performance level, i.e. the level reached in the fifth training session, decreased over the 71 week period, when the platform escape latency and the distance swam to reach the platform, measured as number of line crossings, were considered. In the last experiments of the series, learning curves were no longer seen: the rats did not improve their performance across the acquisition sessions. By contrast, the swimming speed and, in the probe trial, the bias for the quadrant where the platform had been positioned during training, did not change. This indicates that a decrease across experiments occurred predominantly with respect to spatial orientation performance, whereas the motor performance appeared to be unchanged. Explanations for this observation, such as differences in viability between shipments and the possible occurrence of genetic drift, are discussed. PMID- 9331491 TI - Assessing the magnitude of the allocentric spatial deficit associated with complete loss of the anterior thalamic nuclei in rats. AB - The behavioural effects of complete lesions of the anterior thalamic nuclei (ANT), the anterior thalamic nuclei plus the lateral dorsal nucleus (ANT + LD), and fornix (FX) were compared using a series of tests of spatial memory. ALl three lesion groups were found to have an equally severe and long-lasting impairment in the acquisition of a T-maze alternation task when compared with the control animals (COMB SHAM). In Experiment 2, the control animals were able to perform the alternation task when the test trial was started from a different location to the sample trial, so demonstrating that they were able to use allocentric cues in order to differentiate the most recently visited arm. In contrast, all the lesion groups performed close to chance level. In fact, for this condition the ANT / LD group was significantly worse than the FX group. In contrast, none of the lesion groups was impaired on an egocentric discrimination and subsequent reversal task (Experiment 3). The control animals came from two different control procedures, a surgical control sub-group (SHAM) and a group of animals that received injections of N-methyl-D-aspartic (NMDA) into the fornix (NMDA SHAM). There were no differences in the performance levels of the NMDA SHAM group compared with the surgical control group in any of the experiments conducted, so showing that the anterior thalamic lesion effects were not due to non-specific damage to the fornix by NMDA. This series of experiments demonstrated that complete lesions of the anterior thalamic region impair the ability to process allocentric information, and provide evidence for a contribution from the lateral dorsal thalamic nucleus. PMID- 9331492 TI - Effects of weekly or daily exposure to the elevated plus-maze in male mice. AB - The elevated plus-maze is an animal model where the behavioural repertoire of rodents is used to detect effects on anxiety. Repeated testing is a procedural variable where contradictory results have been reported. Some laboratories have found stable test-retest profiles, although other studies have reported reduced open arm exploration. The objective was to further discern behavioural changes in the behaviour of the mouse after either weekly or daily tests. Behaviour was videotaped and later analysed. Behavioural patterns were encoded from an ethological point of view, a nine-pattern ethogram being employed. Other parameters such as percent time on the different sections of the maze were evaluated as well. Descriptive analysis revealed a progressive decrease in percent time spent on open arms (in weekly-tested mice), percent time on central platform, open arm entries, percent open entries, unprotected stretched attention posture (uSAP) and unprotected head-dipping (uDip), together with an augmented number of closed arm returns and percent time spent on closed arms. Taken together, these findings are consistent with an enhanced anxiety level across the tests. It is worth noting that percent time on open arms, a traditional anxiety related parameter, was not progressively decreased in daily-tested mice. Other than expected, exploratory and locomotor elements such as sniffing, rearing, closed arm entries, and total arm entries remained quite similarly elicited throughout the tests, suggesting that locomotor habituation was not developed. However, grooming, considered a displacement response, habituated across the tests. In conclusion, the findings of the present study support the hypothesis that anxiety is enhanced after test repetition, and indicate that test-retest profiles are far from stable, except for exploratory locomotor activity. PMID- 9331493 TI - Fos-like immunoreactivity in forebrain regions following self-stimulation of the lateral hypothalamus and the ventral tegmental area. AB - According to the descending-path hypothesis, the direct excitation of descending fibers linking the lateral hypothalamus (LH) and ventral tegmental area (VTA) contributes to the rewarding effect produced by electrical stimulation of the medial forebrain bundle (MFB). To visualize forebrain neurons activated by stimulation of both the LH and VTA, Fos-like immunoreactivity (FLIR) in forebrain regions was assessed following self-stimulation of these two sites in male rats. Among the regions where FLIR was greater in the stimulated hemisphere following either LH or VTA stimulation were the anterior LH, the substantia innominata, and the bed nucleus of the stria terminalis, and olfactory tubercle. These findings are analyzed with reference to the effects of forebrain lesions on self stimulation of the MFB. Advantages and limitations of using FLIR to identify neurons activated by rewarding stimulation are discussed. PMID- 9331494 TI - Force and the motor cortex. AB - The relation between the activity of cells in the motor cortex and static force has been studied extensively. Most studies have concentrated on the relation to the magnitude of force; this relation is more or less monotonic. The slope of the relation, however, shows considerable variation among different studies and seems to be inversely associated with the range of forces over which the cell activity has been studied. Cells in the motor cortex also show variation in activity with the direction of static force. When both the direction and the magnitude of static force are allowed to vary, a majority of cells show significant changes in activity with direction of force alone, an intermediate number relate to both direction and magnitude, while a small number relate purely to the magnitude. This suggests that the direction of static force can be controlled independently of its magnitude and that this directional signal is especially prominent in the motor cortex. In general, it has been more difficult to study the relations to dynamic force. There is a correlation between motor cortex cell activity and the rate of change of force. The direction of dynamic force is also an important determinant of cell activity. When both static and dynamic force output are required (for example, with arm movement in the presence of gravity) it is the dynamic signal that is most clearly reflected in motor cortex activity. The relations between motor cortex activity and static or dynamic force are not invariant, but may be modified by the behavioral context of the motor output. PMID- 9331495 TI - Functional recovery of porcine hepatocytes after hypothermic or cryogenic preservation for liver support systems. AB - The provision of an immediate supply of isolated porcine hepatocytes for artificial liver support requires preservation techniques that will allow maintenance of cell viability and detoxification functions. By means of a simple and cost-effective cryopreservation system, porcine hepatocytes can be available for both local and distant medical treatment facilities. Additionally, cryopreservation provides an adequate period for quality control testing to be completed prior to use of any specific cell lot. We are reporting a dual approach, namely the preservation of porcine hepatocytes, at 4 degrees C and at 196 degrees C in liquid nitrogen (LN2). Using a combination of cryoprotectant agents with Chee's modified Eagle's culture media (CEM), collagenase isolated hepatocytes stored at 4 degrees C for 24 h maintained 80% of the initial diazepam metabolism measured in freshly isolated cells and nearly 100% of initial function was preserved in hepatocytes stored up to 6 mo at -196 degrees C. University of Wisconsin solution (UW) was also tested and while adequate for 4 degrees C storage, it certainly did not match the performance of the CEM formulations for preservation of metabolic function of cells stored in liquid nitrogen. Based on our results of viability and detoxification function the combination of CEM with DMSO, polyethylene glycol and serum provided optimal protection for LN2 frozen cells. Other findings in these studies underlined the importance of the gradual introduction of DMSO in the prefreezing process, the period of osmotic equilibration, and the rapid postthaw withdrawal of this agent to minimize cytotoxic effects at these critical stages. Our freezing methodology provides the foundation for further technological developments in the cryopreservation of the large numbers of cells (billions) that are necessary for extracorporeal liver assist devices. PMID- 9331496 TI - Selective intraportal transplantation of DiI-marked isolated rat hepatocytes. AB - Transplantation of isolated hepatocytes is a promising alternative to orthotopic liver transplantation in experimental animal models with acute hepatic failure and hereditary enzyme defects. Conventional light microscopy identification of hepatocytes within recipient livers has been limited due to the inability to distinguish between donor and recipient liver cells. In this study, we labeled hepatocytes intracellularly with the fluorescent dye DiI-18 prior to selective intraportal or intrasplenic transplantation. Syngeneic LEW rat hepatocytes were isolated and 2 x 10(7) fluorescence-labeled cells were transplanted by intraportal infusion selectively into 2/3 of the recipient liver lobules to avoid lethal portal hypertension. Rats were sacrificed on postop days 1, 3, 5, 10, 20, and 40. Histological examination was performed using light and fluorescence microscopy counterstained by light green dye. The quantity of transplanted hepatocytes residing within the recipient liver was determined by FACS analysis after enzymatic digestion of the recipient liver lobules. Engrafted hepatocytes were identified in the periportal regions of transplanted liver lobules. The stained hepatocytes were retrieved up to 20 days postop using fluorescent microscopy. Using FACS analysis the number of labeled hepatocytes was found to diminish over time following transplantation from 2.1% on postop day 1 to 0.5% on day 10. Labeled hepatocytes transplanted into the spleen were retrieved in clusters up to 20 days postop (the last day of observation). Furthermore, the migration of labeled hepatocytes from spleen to liver parenchyma was observed following intrasplenic transplantation. However, after selective intraportal transplantation, only fluorescent debris was found in splenic and pulmonary tissue upon examination of various organs. This article describes the method of fluorescent labeling of rat hepatocytes and reports the feasibility and limitations of using DiI-18 as a marker. PMID- 9331497 TI - Highly porous polymer matrices as a three-dimensional culture system for hepatocytes. AB - Hepatocyte-based therapies (e.g., hepatocyte transplantation and extracorporeal support devices) may provide alternative therapies to treat patients with liver disease, but suitable approaches to localize these cells to a given location while maintaining liver-specific gene expression must be developed. The suitability of highly porous three-dimensional sponges fabricated from poly (L lactic acid) [PLLA] as an hepatocyte culture system was evaluated in this study. Sponges were fabricated utilizing a particulate leaching technique, and were approximately 95% porous, with an average pore diameter of 180 microns. Hepatocytes seeded into these sponges adhered and remained viable for 14 days. However, the secretion rate of albumin from these cells, an indication of liver specific gene expression, was low (approximately 6 pg/cell/day at day 1), and decreased steadily over the 14 days of the experiment. Coating sponges with collagen, and more preferably, immobilizing cells within the PLLA sponges with a collagen gel, led to enhanced cell survival and albumin secretion at all time points. These data suggest that porous PLLA sponges may provide a novel system for long-term culture of hepatocytes, and proper design of the system may allow the liver-specific gene expression of hepatocytes transplanted in these matrices to be enhanced. PMID- 9331498 TI - Polymer-encapsulated PC-12 cells demonstrate high-affinity uptake of dopamine in vitro and 18F-Dopa uptake and metabolism after intracerebral implantation in nonhuman primates. AB - Intracranial implantation of polymer-encapsulated PC-12 cells has been shown to improve motor behavioral performance in animal models of Parkinson's disease. The purpose of this blinded study was to examine whether such improvement is associated with the active uptake and metabolism of dopamine precursors by intracerebrally implanted polymer-encapsulated PC-12 cells. In an in vitro experiment we demonstrate that 3H-dopamine uptake by PC-12 cells was 10(8) fmol/min x 10(6) cells, and that this uptake can be specifically blocked 88% by the addition of 10nM of nomifensine. In the in vivo experiments, polymer encapsulated PC-12 cells were implanted in four MPTP-treated monkeys into the left deep parietal white matter (R1) or left striatum (R2-4). A fifth MPTP treated monkey (R5) served as a control and received left striatal implants of empty capsules. 18-F-Dopa Positron Emission Tomography (PET) imaging was performed on each monkey before and after implantation surgery by blinded investigators. PET images obtained 5-13 wk after implantation demonstrated well delineated focal areas of high 18F-dopa uptake in R1, R2, and R4. The focal area of high 18F-dopa uptake in R1 precisely coregistered on a brain magnetic resonance image to the site of implantation. R3 (in whom the polymer-encapsulated PC-12 cells demonstrated poor cell survival upon explantation) and R5 (empty capsules) failed to demonstrate any area of increased 18F-dopa uptake in their PET images. Histological examination of the host brain revealed no sprouting of dopaminergic nerve terminals around the implantation sites of the polymer encapsulated PC-12 cells. These results indicate that the previously noted behavioral improvement after intrastriatal implantation of polymer encapsulated PC-12 cells is at least in part due to their highly specific uptake and metabolism of dopamine precursors. Furthermore, these data suggest that polymer encapsulated PC-12 cells can store, reuptake, and functionally replenish dopamine and therefore, may be an effective treatment for Parkinson's disease. PMID- 9331500 TI - Bone marrow mononuclear cell count does not predict neutrophil and platelet recovery following autologous bone marrow transplant: value of the colony-forming unit granulocyte-macrophage (CFU-GM) assay. AB - The common use of the marrow autograft mononuclear cell (MNC) count derives from positive correlative studies following allogeneic transplantation and from earlier conflicting data regarding the value of the bone marrow autograft colony forming unit granulocyte-macrophage (CFU-GM) assay for prediction hematologic recovery after ABMT. We conducted a retrospective analysis at our institution to determine whether autograft CFU-GM levels predict engraftment of neutrophils and platelets after ABMT in heavily pretreated patients with hematologic malignancies. Between 1 January 1993 and 1 March 1995, 58 heavily pretreated patients received only marrow cells as the autograft product. Patients with Hodgkin's disease (n = 25), acute myeloid leukemia (n = 19), and non-Hodgkin's lymphoma (n = 14) underwent intensive therapy with etoposide and melphalan. Unpurged marrow containing a minimum of 1.5 x 10(8)/kg (range: 1.5-4.8) was infused. Median time to an absolute neutrophil count > or = 0.5 x 10(9)/L was 21 days (range 10-270) and median time to a platelet count > or = 20 x 10(9)/L independent of transfusions was 44 days (range 13-317). There was no correlation between autograft MNC count and neutrophil or platelet engraftment. However, a correlation between autograft CFU-GM and both platelet and neutrophil recovery was demonstrated with a threshold CFU-GM of 3 x 10(4)/kg; delayed neutrophil recovery was observed in 79% of patients below this threshold compared to only 9% in those with an autograft CFU-GM level of more than 3 x 10(4)/kg (p = 0.0001). Similarly, platelet recovery was delayed in 76% of patients below, and 20% of those above this threshold (p = 0.003). We conclude that marrow autograft CFU-GM is predictive of engraftment of both platelets and neutrophils in heavily pretreated patients after ABMT for hematological malignancies. PMID- 9331499 TI - Organization and histochemical phenotype of human fetal cerebellar cells following transplantation into the cerebellum of nude mice. AB - Previous rodent studies have demonstrated the capacity of cerebellar transplants to organize into trilaminar cell layers typically observed in the normal cerebellum. In Purkinje Cell (PC)-deficient animals, PCs will migrate into the host and form synaptic connections. Recently, fetal cerebellar grafts transplanted into the Purkinje cell degeneration (pcd) mutant mouse were shown to result in an improvement of motor behaviors. These studies indicate the potential therapeutic use of neural transplantation in patients with cerebellar degeneration. In the present study, human fetal cerebellar tissue (8.5 wk postconception) was dissociated and transplanted into the normal cerebellum of nude mice. Six months following transplantation, histological analysis revealed donor cells in recipient mice. Immunostaining for the 28 kDa calcium-binding protein (calbindin) revealed the presence of donor PCs that were organized in discrete cellular layers within the transplant neuropil. In most cases the dendritic processes were oriented in a planar fashion perpendicular to the transplant cell layer. Human neurofilament immunostaining revealed bundles of donor fibers within the core of the transplant and/or at the periphery. These bundles were found to be calbindin positive (PC fibers). Three animals provided evidence of donor PC axon growth ventrally into host white matter, and in one case, this ventral migration reached the deep cerebellar nuclei. Most notable was the development of a pronounced folia-like organization by the implanted cell suspensions. Glial processes within the grafts were aligned perpendicular to the long axis of the transplant folia. These results demonstrate the capacity of human fetal cerebellar cell suspension to reorganize into cell layers typical of the normal cerebellum following transplantation into the rodent cerebellum, and develop an organotypic folia-like organization. PMID- 9331501 TI - Glucoregulation after canine islet transplantation: contribution of insulin secretory capacity, insulin action, and the entero-insular axis. AB - The physiological glucoregulatory mechanisms after islet transplantation have been incompletely investigated. We studied the insulin secretory capacity (ISC) by intravenous arginine stimulation during 35-mM glucose clamps, insulin action during hyperinsulinemic euglycemic clamps, and mixed-meal stimulation at 6-9 mo after intrasplenic islet autotransplantation in 8 dogs, as compared with 30 controls. The enteroinsular axis in the recipients was examined by infusion of porcine glucose-dependent insulinotropic polypeptide (GIP) and human glucagon like peptide-1 (GLP-1) (7-36 amide) under 8.5-mM glycemic clamp conditions in order to mimic the postprandial glycemia after transplantation. The grafts comprised 25% of the native islet mass, and the ISC likewise averaged 25% of the control value. The postprandial insulin response, in contrast, had increased to 140% after transplantation--albeit with a concomitant glucose excursion to approximately 8.5 mM. Insulin action declined on average by 45% posttransplant. The ISC correlated both with the postprandial glucose excursion and insulin action in the grafted dogs. Insulin action did not correlate with the postprandial glucose excursion. Infusion of GIP had no effect, but GLP-1 nearly doubled glucose-stimulated insulin. Thus, a hyperglycemia-enhanced insulinotropic effect of GLP-1, and perhaps other gut hormones, may account for the difference in the insulin response to the intravenous and oral challenges. Because the ISC reflects the engrafted islet mass and appears to be the primary determinant of glucose tolerance, transplantation of higher islet doses should allow prolonged near-normal glucoregulation--at least in the autotransplant setting. PMID- 9331503 TI - Evaluation of intracerebral grafting of dopamine-secreting PC12 cells into allogeneic and xenogeneic brain. AB - The PC12 phenochromocytoma tumor cell line is derived from a rat adrenal medullary tumor and secretes dopamine. We have previously reported that grafted microencapsulated PC12 cells using agarose and poly (styrene sulfonic acid) survived in the xenogeneic brain without immunosuppression. To investigate whether unencapsulated PC12 cells form a tumor and how they provoke immunological reaction, PC12 cell suspension was implanted into the striatum of Sprague-Dawley rat (allogeneic graft) or guinea pig (xenogeneic graft) and histological analysis using Nissl stain and immunocytochemical analysis using antityrosine hydroxylase (TH) antibody were performed 1, 2, and 4 wk after transplantation. Host animals were not immunosuppressed. PC12 cells formed a mass 1 and 2 wk after transplantation both in allogeneic and xenogeneic brain. These grafted PC12 cells were immunoreactive to anti-TH antibody. Four weeks after transplantation, however, grafted PC12 cells in the allogeneic brain were only found within the restricted area near the site of implantation. In the xenogeneic brain, only the trace of grafted PC12 cells were found around the site of implantation 4 wk after transplantation. In both allogeneic and xenogeneic animals, a number of lymphocytes were found in and around the grafts at all period investigated. These findings indicate that PC12 cells could survive in the allogeneic or xenogeneic brain for 2 wk and were ultimately rejected by immunological reaction by 4 wk after transplantation. Implantation of encapsulated PC12 cells in the allogeneic or xenogeneic brain is considered a safe and effective method for delivering dopamine into the brain because PC12 cells will not form a tumor in the long-term even if capsules are damaged in some reason. PMID- 9331502 TI - CTLA4-Ig prolongs survival of microencapsulated neonatal porcine islet xenografts in diabetic NOD mice. PMID- 9331504 TI - Implantation of xenogeneic transgenic neural plate tissues into parkinsonian rat brain. AB - Xenografting must be considered as a means of establishing neural transplantation therapy and of securing fetal neural tissues as donor material. The early stage (embryonic day 8.5, E8.5) embryonic mesencephalic neural plate (NP) from transgenic mice was examined for possible application in effective xenografting therapy. As recipients, Parkinsonian rats treated with 6-hydroxydopamine were used, and as donors, GT4-2 mice into which a beta-galactosidase gene was introduced to allow brain tissue differentiation from the recipients by X-gal staining. Three microscopic pieces of E8.5 GT4-2 mice NP were injected into the striatum of the Parkinsonian rats. Some hosts were given immunosuppressants (cyclophosphamide and FK506) (IS group), others were not (non-IS group). Amphetamine-induced rotation was examined at days 11 and 21 after grafting (D11 and D21, respectively), and morphological investigations were performed using hematoxylin-eosin (H-E), X-gal, and thyrosine hydroxylase (TH) staining. The rotations were counted in 30 of the 38 transplanted rats before and after grafting. Histological data were obtained from 19 of these 30 rats. In 11 of them the grafts survived (survival group) and in the remaining 8, the grafts were unsuccessful (rejection group). In the survival group at D11, the mean number of rotations made by transplanted rats expressed as a percentage of the number before grafting (rotation percentage) decreased to 43.8% (n = 9), which, in comparison with the average of 125.9% (n = 6) in the rejection group, reveals significant behavioral recovery (p < 0.01). The rotation percentage at D21 was 23.8% in the survival group (n = 4) and 84.5% in the rejection group (n = 3). Behavioral recovery was thus seen to improve with time in the survival group. In the IS group (n = 19), the rotation percentages averaged 74.9% (D11, n = 15) and 51.1% (D21, n = 7), while the non-IS group averages were 136.7% (D11, n = 9) and 140.7% (D21, n = 9), indicating a tendency for better behavioral recovery in the IS group than in the non-IS group (p < 0.05). Fifteen IS group rats were studied histologically, 10 (sacrificed on D11, D21) from the survival group and 5 (sacrificed on D11, D21) from the rejection group, In the non-IS group (n = 4), there was a graft in only one rat sacrificed on D11. There were many X-gal positive and TH positive cells in the grafts, suggesting that mouse NP survived, and differentiated into TH positive neurons in the rat brain. Xenografted NP has the potential to cure central nervous system diseases. PMID- 9331505 TI - Efficiency of neural differentiation of mouse P19 embryonal carcinoma cells is dependent on the seeding density. AB - Serum-free culture conditions for retinoic acid-induced neural differentiation of mouse P19 embryonal carcinoma cells were determined for future ex vivo retroviral gene transfer and brain transplantation studies. Neural differentiation of P19 cells was dependent on the seeding densities, and both neurons and astroglia differentiated efficiently at high seeding densities (2 x 10(4) and 5 x 10(4) cells/cm2) but not at low seeding density (1 x 10(4) cells/cm2). In addition, P19 cells cultured at 5 x 10(4) cells/cm2 showed neural differentiated whether or not they were infected with Friend leukemia virus FrC6-V, which inhibited neural differentiation at 2 x 10(4) cells/cm2. These results indicate that FrC6-V infected P19 embryonal carcinoma cells should be seeded at high density to achieve efficient neural differentiation in vitro for ex vivo gene transfer with a FrC6-V-derived retroviral vector system. PMID- 9331506 TI - Transplantation of xenogeneic cells secreting beta-endorphin for pain treatment: analysis of the ability of components of complement to penetrate through polymer capsules. AB - The permeation of component of complement and secreted peptides through polymer capsules (PM30, K6305, and K5708) were examined. To analyze permeability by complement, the degree of hemolysis of sensitized sheep erythrocytes (EA) (1 x 10(9)/ml) enclosed in each type of capsule was examined after 24-h incubation in culture medium containing 10% human serum. PM30 and K6305 prevented the permeation of complement well, while K5708 did not. EA suspended in alginate prevented hemolysis even in K5708. Peptide permeation through the capsules was assessed by measuring the concentration of ACTH secreted by proopiomelanocortin (POMC)-gene-transfected-Neuro2A in the culture medium on days 4, 7, 14, 21, and 28 after encapsulation. The ACTH levels in the culture medium remained high until day 28. Alginate appeared to prevent the secretion, because ACTH levels decreased in alginate-suspended cells after day 14. The PM30-K6306 double capsules containing cell lines, Neuro2A, BHK21 (hamster fibroblasts), L929 (mouse fibroblasts), and HF-SKFII (human fibroblasts) were transplanted into the cerebrospinal fluid (CSF) space of the monkeys in the lumber region. The morphological examination showed the partial survival of Neuro2A, and BHK21 and HF-SKFII, which were cells concordant with the monkeys. On the other hand, L929 cells, which were discordant with the monkeys, could not survive at all. Because these results suggest that the complement components penetrate the polymer capsules, concordant cells are preferable for xenografting with polymer capsules into the CSF space. PMID- 9331507 TI - Enzymatic activity and expression of cytochrome P450 LA omega within intrasplenically transplanted fetal hepatocytes in spontaneously hypertensive rats. AB - We examined the expression and enzymatic activity of cytochrome P450 LA omega within transplanted hepatocytes. Fetal hepatocytes were harvested at day 20 of gestation from spontaneously hypertensive rats (SHRs) and transplanted into recipient adult SHR spleens. Microscopic examination of the recipient spleens at 4 and 10 wk after transplantation revealed masses of hepatocytes with cord-like structures in the red pulp. Immunochemical studies detected cytochrome (cyto) P450 LA omega in the fetal hepatocytes before transplantation without prior induction. Although the cyto P450 LA omega was not detected by the second week after transplantation, by the 6th and 10th wk after transplantation, it was. Cyto P450-arachidonic acid omega/omega-1 hydroxylase activity (formation of 20- and 19 hydroxyeicosatetraenoic acid) was detected at 10 wk after transplantation, but not 2 or 6 wk after transplantation. These results demonstrated that fetal hepatocytes can be transplanted successfully into recipient spleens and then grow in the spleens, as in the case of the adult hepatocyte response. PMID- 9331508 TI - Immunoregulation via adhesion molecules in allogenic and xenogenic hepatocyte transplantation to Nagase's analbuminemic rats. AB - We investigated the effects of monoclonal antibodies (mAbs) against lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM 1) on intrasplenic allogenic and xenogenic hepatocyte transplantation (HCTx) to analbuminemic rats. Ten to 12-wk-old male Nagase's analbuminemic rats (RT1l) were used as recipients, Wistar/Shi rats (RT1k) were used as donors for allografts and BALB/C mice were used as donors for xenografts. The experimental groups were as follows: group 1, allo-HCTx (n = 7); group 2, allo-HCTx + antirat ICAM-1/antirat LFA-1 mAbs (1.0 mg/kg/day, for 7 days, respectively) (n = 6); group 3, xeno-HCTx (n = 5); group 4, xeno-HCTx + mAbs (antimouse LFA-1/antirat ICAM-1) (n = 5). group 5, xeno-HCTx + mAbs (antirat LFA-1/antimouse ICAM-1) (n = 5). Serum rat albumin levels were measured in groups 1 and 2, and serum mouse albumin levels were measured in groups 3, 4, and 5, as indicators of the function of grafted hepatocytes. In allotransplantation groups, the serum rat albumin levels in the mAbs-treated group were significantly higher than those in the control group for 6 wk after transplantation. In xenotransplantation groups, no increase in the serum mouse albumin levels was detected in any group PMID- 9331509 TI - An attempt to add biological functions by genetic engineering in order to produce high-performance bioreactor cells for hybrid artificial liver: transfection of glutamine synthetase into Chinese hamster ovary (CHO) cell. AB - In the course of immortalization, hepatocyte cell lines lose their original differentiated functions, such as ammonia removal and urea formation, drug metabolism, serum protein synthesis, etc. (Enosawa et al., Cell Transplant. 5:S39 S40; 1996). With the aim of adding lost or deficient functions and producing cell lines for the bioreactor of a hybrid artificial liver, rat glutamine synthetase (GS) gene was transfected into Chinese hamster ovary (CHO) cells, because it is able to lower the ammonia level. The GS gene-inserted pSV2 plasmid was transfected into the CHO-K1 line by electroporation. Transfected CHO (GS-CHO) cells were cultured in a glutamine-free medium containing ammonia, glutamic acid, and the GS inhibitor methionine sulfoximine (MSX). The MSX concentration was increased stepwise from 25 mumol/L to 1600 mumol/L to amplify the GS gene. In several GS-CHO sublines resistant to 300-1600 mumol/L of MSX, the specific activities of GS were increased from 0.2 x 10(4) to 1.7-2.9 x 10(4) unit/10(6) cells. When the amplified GS-CHO cells were cultured in the ammonia-containing medium, a slow but steady decrease of the ammonia level was observed when the level was high. Finally, the prospect of genetically modulated cells for bioreactors in the hybrid artificial liver is discussed. PMID- 9331510 TI - Subcutaneous xenotransplantation of hybrid artificial pancreas encapsulating pancreatic B cell line (MIN6): functional and histological study. AB - The biohybrid artificial pancreas is designed to enclose pancreatic tissues with a selectively permeable membrane that immunoisolates the graft from the host immune system, allowing those endocrine tissues to survive and control glucose metabolism for an extended period of time. The pancreatic B cell line MIN6 is established from a pancreas B cell tumor occurring in transgenic mice harbouring the human insulin promoter gene connected to the SV40 T-antigen hybrid gene. It has been proven that glucose-stimulated insulin secretion in MIN6 cells retains a concentration-dependent response similar to that of normal islets. In this study, we performed the histological and functional examination of three-layer microbeads employing MIN6 cells after subcutaneous xenotransplantation to evaluate this device as bioartificial pancreas. MIN6 cells were microencapsulated in three-layer microbeads formulated with agarose, polystyrene sulfonic acid, polybrene, and carboxymethyl cellulose. Microbeads were xenogenically implanted in the subcutaneous tissue of the back of Lewis rats with streptozotocin-induced diabetes. One week after implantation, microbeads were retrieved and cultured for 24 h before the static incubation. There was no evidence of adhesion to the graft and the fibrosis in the transplantation site as determined by gross visual inspection. Microscopic examination demonstrated that retrieved microbeads maintained normal shape, containing intact MIN6 cells. Histological study showed that these MIN6 cells in the microbeads appeared to be viable without cellular infiltration within or around the microbeads. Immunohistochemical analysis of the microbeads clearly revealed the intense staining of insulin in the cytoplasm of encapsulated MIN6 cells. Insulin productivity of MIN6 cells in the microbeads is strongly suggested to be preserved. In response to 16.7 mM glucose stimulation, static incubation of microbeads 1 wk after implantation caused the 2.3 times increase in insulin secretion seen after 3.3 mM glucose stimulation (84.3 +/- 10.0 vs. 37.4 +/- 10.7 microU/3 x 10(6) cells/hr, n = 5 each, p < 0.01). This study demonstrates that three-layer microbeads encapsulating MIN6 cells retain excellent biocompatibility and maintain good insulin secretion even after subcutaneous xenotransplantation, suggesting the possible future clinical application of this unique bioartificial pancreas to subcutaneous xenotransplantation. PMID- 9331511 TI - Prolonged effect of troglitazone (CS-045) on xenograft survival of hybrid artificial pancreas. AB - Troglitazone (CS-045), a thiazolidinedione derivative, is a new oral antidiabetic agent that enhances insulin sensitivity and improves insulin responsiveness. In this study we examined the effects of CS-045 on the survival of xenografted bioartificial pancreas. Isolated rat islets were microencapsulated with three layer agarose microcapsules (polybrene, carboxymethyl cellulose, and an agarose polystyrene sulfonic acid mixture). Diabetes was induced by intraperitoneal injection of streptozotocin 220 mg/kg. Recipient diabetic mice were separated into two groups. In the CS-045 treated group, the recipient mice were given feed mixed with CS-045 (0.2% w/w) starting from 1 wk before transplantation up to graft failure. The mice in the control group had feed without CS-045. Three hundred microencapsulated rat islets were xenotransplanted into the intraperitoneal cavity of each recipient mouse in both groups. One month after xenotransplantation, IVGTT was performed for all recipients. Xenotransplantation of 300 rat islets in microcapsules decreased the nonfasting blood glucose levels of both groups within 2 days. In the CS-045-treated group (n = 3), the normoglycemic period lasted for more than 1 mo without administration of immunosuppressive drugs (45 +/- 4.3 days). However, in the control group (n = 4), the blood glucose levels of all recipients were already elevated on day 4. In the IVGTT study, the glucose assimilation was markedly and significantly better in the CS-045-treated group than in the control group (K = 1.7 +/- 0.1 vs. 0.7 +/- 0.28 respectively, p < 0.01). This study demonstrates that a newly developed oral antidiabetic agent, CS-045 could favorably ameliorate the diabetic state of the recipients xenotransplanted with the bioartificial pancreas, leading to an improved glucose tolerance and longer xenograft survival. PMID- 9331513 TI - Behavioral approaches to the functional assessment of NMDA-mediated neural transmission in intact mice. AB - Altered neurotransmission mediated by L-glutamate at the level of the N-methyl-D aspartic acid (NMDA) receptor complex has been implicated in the pathophysiologic mechanisms of several major neuropsychiatric disorders. Moreover, strategies for the pharmacologic manipulation of NMDA-mediated neural transmission have been discussed for the treatment of disorders as diverse as schizophrenia, seizures, stroke, and traumatic brain injury, MK-801, an uncompetitive allosteric antagonist of the NMDA receptor complex, was shown to antagonize electrically precipitated seizures in a dose-dependent manner and elicit popping behavior in mice. Changes in the ability of MK-801 to antagonize electrically precipitated seizures or elicit popping behavior caused by stress or pharmacologic manipulations may reflect alterations in the populations of NMDA-associated channels responsible for these behavioral actions (e.g., the number of them in the open configuration or their size, shape, and charge characteristics). We used these paradigms to study the pharmacologic actions of an allosteric glycinergic intervention (i.e., milacemide), inhibitors of the "nitric oxide cascade" (i.e., 7-nitroindazole and methylene blue), and conventional (i.e., haloperidol) and atypical (i.e., clozapine) antipsychotic medications on NMDA-mediated neurotransmission in the intact mouse. Also, marked differences in the ability of MK-801 to elicit popping behavior in inbred mouse strains suggest that they differ in their populations of NMDA receptor complexes responsible for mediating this behavior. This latter observation could lend itself to the identification of specific genetic loci contributing to this behavior. In view of the ability of phencyclidine (PCP) to precipitate a schizophreniform psychosis and the action it shares with MK-801 on NMDA-mediated neurotransmission, the characterization of these genetic loci in mice may inform the search for human loci responsible for the susceptibility to "PCP-psychosis" and schizophrenia. PMID- 9331512 TI - Microchimerism and hyporesponsiveness induced by intraportal injection of donor spleen cells in rats. AB - It is controversial whether or not microchimerism (MC) is responsible for the induction and maintenance of donor-specific tolerance. We have shown that intraportal injection (i.p.) of donor splenocytes induces a long-term graft survival of liver and heart in rats. In this study, we examined by polymerase chain reaction (PCR) the status of MC in the liver, spleen, and blood of rat cardiac recipients following i.p. or intravenous injection (i.v.) of donor splenocytes. Male DA (RT1a) and Wistar (RT1k) rats were used as donors and recipients, respectively. Heterotopic heart transplantation was performed 10 days after i.p. or i.v. injection of 5 x 10(7) DA spleen cells. DA cardiac allografts were rejected with a mean survival time (MST) of 11.9 +/- 1.6 (n = 10) days in nontreated recipients. Injections (i.v.) led to no significant prolongation of graft survival (MST: 11.2 +/- 1.9 days, n = 6), while i.p. or i.v. injection alone resulted in significant MC in these organs throughout the observation time over 60 days. MC was detected in the spleen, liver, and blood of cardiac recipients 7 days after transplantation and also even after cessation of cardiac heartbeat 21 days after transplantation. This was the case with either i.p. or i.v. group, which showed MC on day 7 after transplantation and persistent MC after cessation of the heartbeat. These data suggests that the presence of MC in the liver, spleen and blood of transplant recipients may not be responsible for immunological unresponsiveness to donor antigens. PMID- 9331514 TI - Side effects of high-dose intravenous immunoglobulins. AB - Intravenous immunoglobulins (IVIgs) are used increasingly as therapy for neuroimmunologic and other autoimmune diseases. With broader use, the number of reported side effects also is growing. Here we review the literature on adverse reactions reported after administration of i.v.Igs. Despite a few recent reports about a high frequency of complications of IVIgs, by and large they can be considered as safe. Mild and self-limited side effects may occur, but severe complications are rare and often associated with other risk factors for these complications. A careful screening for preexisting illnesses and monitoring of some laboratory parameters can minimize the risks of IVIg therapy. PMID- 9331515 TI - The long-duration action of levodopa may be due to a postsynaptic effect. AB - A single dose of levodopa (L-DOPA) reduces motor disability in Parkinson's disease (PD) for a few hours, a short-duration effect. However, there are suggestions that L-DOPA may also produce a long-duration benefit of some days. In the present study, we examined the long-duration action of L-DOPA by observing the time taken to achieve maximum stable benefit after starting a constant dose of sinemet-CR (sinemet-CR) (200 g L-DOPA/50 mg carbidopa) twice daily in nine newly diagnosed patients, and the time taken to deteriorate back to baseline after stopping treatment. A single dose of sinemet-CR (200 mg L-DOPA/50 mg carbidopa) had little obvious short-duration action on the Unified PD Rating Scale (UPDRS) motor scores in the majority of patients, either before starting chronic sinemet-CR therapy (200 mg L-DOPA/50 mg carbidopa, b.i.d.) or after chronic treatment. However, all patients gradually improved on chronic sinemet-CR therapy, taking 9.3 +/- 1.8 days to achieve maximum response. On stopping chronic sinemet-CR treatment, it took 6.8 +/- 3.0 days for the same patients to deteriorate back to baseline motor disability. In similar experiments, the time taken to deteriorate back to baseline after stopping treatment with the directly acting dopamine agonist ropinirole (9-21 mg daily) in eight other de novo patients with PD was found to be 6.2 +/- 1.7 days. The long-duration effect of L DOPA and ropinirole may, therefore, be due to some slowly evolving postsynaptic pharmacodynamic change in the central nervous system (CNS). Loss of this long duration action may be responsible for the emergence of motor fluctuations on chronic L-DOPA therapy. PMID- 9331516 TI - Movement characteristics in Parkinson's disease: determination of dopaminergic responsiveness and threshold. AB - We evaluated the responsiveness of tap rate (TR), movement time (MT), and reaction time (RT) to intravenous (i.v.) (n = 10) and subcutaneous (s.c.) (n = 16) administration of apomorphine in patients with Parkinson's disease (PD). In the second part of this study, we evaluated the feasibility of the commonly used 15% TR threshold, above which a patient is considered to be a responder. Compared to MT, TR emerged as the most responsive measure of bradykinesia during both i.v. and s.c. administration of apomorphine. RT showed no response to dopaminergic stimulation. To evaluate the influence of threshold on the number of responsive sessions, we determined the baseline variability of TR by means of the coefficient of variation (CV) in 39 patients with PD. Our results show approximately similar numbers of responsive sessions using the 15% and 2CV threshold. Our findings suggest that a simple repetitive motor task--TR--is more responsive than is the MT task. Finally, the 15% threshold may be considered an adequate threshold for TR in assessment of dopaminergic responsiveness of bradykinesia. PMID- 9331517 TI - Relationship between levodopa concentration, dyskinesias, and motor effect in parkinsonian patients: a 3-year follow-up study. AB - We conducted a 3-year prospective assessment of the relationship between levodopa (L-DOPA) plasma concentration and both dyskinesias and tapping motor response after a standard oral L-DOPA dose in 11 parkinsonian patients, Hoehn & Yahr (H & Y) stages 2-3, with L-DOPA therapeutic response complicated by involuntary movements. Over the 3-year period, duration of the tapping effect significantly decreased, while that of dyskinesias showed minor changes. Initially, the L-DOPA therapeutic response significantly outlasted the dyskinesia effect and progressively shortened to parallel the dyskinesia profile at the more advanced clinical stage. According to kinetic-dynamic modeling, L-DOPA concentrations producing 50% of maximum therapeutic and toxic effects (EC50) also changed independently. EC50 for dyskinesias did not vary over time. EC50 for the tapping effect was, at the first observation, significantly lower than that of the matched value for dyskinesias and progressively rose to values similar to the EC50 for dyskinesias by the third year of follow-up. These data suggest a dissociation of the kinetic-dynamic relationship of L-DOPA motor and dyskinesia effects, possibly reflecting different cerebral handling of exogenous levodopa derived dopamine with disease progression. PMID- 9331519 TI - Vitamin E treatment in tardive dystonia. AB - Tardive dystonia is a disorder characterized by abnormally sustained posturing associated with the use of dopamine-receptor blocking agents such as antipsychotic drugs. However, the structural pathologic and pathophysiologic features of this disorder are unknown, and no consistently effective pharmacologic treatment is available. Patients with tardive dystonia mostly are young men. We present the case of one substantially improved with treatment by 1200 mg/d (IU) of vitamin E. PMID- 9331518 TI - Lack of adverse interactions between concomitantly administered selegiline and citalopram. AB - We have evaluated the risk for pharmacokinetic and/or pharmacodynamic interactions of concomitantly administered selegiline, a selective monoamine oxidase type B inhibitor, and citalopram, a widely used selective serotonin uptake inhibitor antidepressant. Two parallel groups of healthy volunteers received 20 mg of citalopram (n = 12) or placebo (n = 6) once daily for 10 days in a randomized, double-blind fashion, followed by concomitant selegiline 10 mg once daily for 4 days. The safety of this drug combination was assessed by measurements of blood pressure, heart rate, body temperature, and inquiries for adverse events. Blood samples were taken for the analysis of serum concentrations of both study drugs and their metabolites and plasma prolactin, adrenaline, noradrenaline, and 3,4-dihydroxyphenylglycol (DHPG); urinary excretion of serotonin and 5-hydroxyindoleacetic acid (5-HIAA) were assessed. After a 5-week washout, the 12 subjects who took citalopram in the first part of the study received 10 mg of selegiline once daily for 4 days to compare the pharmacokinetics of selegiline with and without concomitant citalopram. The safety analysis showed no significant differences in vital signs or the frequency of adverse events between the study groups. Plasma prolactin concentrations were increased by 40% after 10 days' treatment with citalopram (p = 0.03); this was not potentiated by concomitantly administered selegiline. The comparison of plasma concentrations of noradrenaline, adrenaline, and DHPG and the amount of serotonin and 5-HIAA excreted into urine between the study groups indicated no signs of subclinical pharmacodynamic interaction between selegiline and citalopram. The relative bioavailability of selegiline was slightly reduced (by 29%; p = 0.008) when citalopram was coadministered compared with selegiline alone. However, no indication of a pharmacokinetic interaction was found in the analysis of serum concentrations of the three main metabolites of selegiline. The pharmacokinetics of citalopram remained unaffected by concomitant selegiline. The present results indicate lack of clinically relevant pharmacodynamic or pharmacokinetic interactions between selegiline and citalopram. PMID- 9331520 TI - Use of antiepileptic drugs in nontraumatic neurosurgical procedures. Is there any best route and time of administration? AB - We assessed in 15 consecutive patients the best route and time of administration for phenytoin (PHT) prophylaxis in neurosurgical procedures. We also correlated PHT levels in serum and cerebrospinal fluid after oral and parenteral loading doses. The mean PHT level was 13.9 micrograms/ml in serum and 2.03 micrograms/ml in cerebrospinal fluid (CSF), with a significant correlation between levels in both compartments (r = 0.73, p < 0.01). Mean PHT levels among the different groups were not statistically significant. We conclude that therapeutic levels of PHT in CSF can be achieved independently of the route of administration, as long as accepted loading doses are used. PMID- 9331521 TI - Reduction of aggressiveness and impulsiveness during clozapine treatment in chronic neuroleptic-resistant schizophrenic patients. AB - Aggressive and impulsive behavior is frequently observed in schizophrenic patients. Previous studies suggest that impulsive aggression may be the most common behavioral correlate of central serotonergic system dysfunction. This study was aimed to determine if clozapine, an atypical antipsychotic agent with potent serotonergic antagonistic properties, can reduce impulsiveness and aggression in neuroleptic-resistant chronic schizophrenic patients. Fourteen neuroleptic-resistant chronic schizophrenic patients were treated with clozapine and prospectively evaluated for aggressiveness and impulsiveness for 18 weeks. Clozapine treatment induced a marked decrease in impulsiveness (32% on the Impulsivity Scale; p < 0.0001) and aggressiveness (98% on the Overt Aggression Scale; p < 0.0001). We conclude that clozapine treatment may be effective in reducing psychotic symptoms as well as in controlling aggressive and impulsive behavior in neuroleptic-resistant chronic schizophrenic patients. PMID- 9331522 TI - Sensitivity of cyclic alternating pattern to prolonged pharmacotherapy: a 5-week study evaluating zolpidem in insomniac patients. AB - To test the effects of a hypnotic drug administered on a regular basis, six adults (four women and two men; mean age 37 years) who complained of transient or short-term insomnia, took zolpidem 10 mg at bedtime for 4 consecutive weeks. The period of active treatment, preceded by a baseline placebo night, ended with a 4 night gradual tapering phase followed by 3 nights of placebo administration. After adaptation to the sleep laboratory, all subjects underwent five polysomnographic recordings; baseline placebo (night 1); 1st, 7th and 28th nights of hypnotic medication (nights 2, 3, and 4 respectively); 3rd placebo night after complete tapering (night 5) and completed a morning visual analogue scale (VAS) for evaluating sleep quality. The sleep recordings were scored according to the conventional procedures (macrostructure) and to the cyclic alternating pattern (CAP) rules (microstructure). Data analysis was based on a repeated-measures analysis of variance integrated by post-hoc comparisons. At the macrostructural level, significant overall modifications (p < 0.035) emerged only from slow-wave sleep (SWS). The amounts of SWS were enhanced along the entire drug period, but a significant difference was found only between the baseline night (10%) and the first night of drug administration (20%). At the microstructural level, CAP rate (the ratio of CAP time to non-rapid eye movement sleep time) showed overall significant variations throughout the entire protocol period (p < 0.0001). Compared with baseline night 1 (59%), the CAP rate was significantly lower on drug nights 2 (32%), 3 (37%), and 4 (38%). The increase in the CAP rate found on night 5 (43%) was still significantly below the baseline value. The VAS scores showed significant overall changes (p < 0.0001), with improved values during the active treatment period. PMID- 9331524 TI - The treatment of comorbid premature ejaculation and panic disorder with fluoxetine. AB - Premature ejaculation is a common sexual disturbance among men. Both open-label and double-blind studies have demonstrated the effectiveness of serotonergic medications for this disorder. These studies support the hypothesis that the serotonergic system has an important role in the modulation of sexual response, especially attainment of orgasm. Serotonergic dysfunction also has been linked to the pathogenesis of panic disorder. Several studies have demonstrated the efficacy of serotonergic drugs in this disorder. The purpose of the present study was to examine the efficacy of fluoxetine, a serotonin selective reuptake inhibitor for the treatment of comorbid premature ejaculation and panic disorder, in 10 men in an open-label design. The patients were given 20 mg of fluoxetine for 8 weeks of the study. Parameters pertaining to sexual function and measures of anxiety were examined. Improvement of premature ejaculation was noted as of week 2 of the study, whereas measures of panic and sexual satisfaction became statistically significant only as of week 4. Further studies with larger samples and longer periods of follow-up are needed in order to determine the usefulness of fluoxetine for the treatment of comorbid premature ejaculation and panic disorder. PMID- 9331523 TI - Clinical and pharmacokinetic evaluation of L-dopa and cabergoline cotreatment in Parkinson's disease. AB - Previous investigations on the mutual pharmacokinetic influence of L-dopa and dopamine agonists in Parkinson's disease (PD) have shown controversial results. Two studies of the possible clinical and pharmacokinetic interaction between L dopa and cabergoline were performed in 10 patients with de novo PD and 12 patients with fluctuating PD. In the first study (de novo patients), cabergoline was administered at increasing dosages until the maximum dosage of 2 mg/day once a day for 8 weeks; subsequently L-dopa (250 mg/day) was added. Blood levels of cabergoline were assayed in two different days, before starting L-dopa, and 1 week thereafter. In the second 8-week study (fluctuating patients), cabergoline was added to the current L-dopa therapy (maximum dosage 4 mg/day once a day). Blood levels of L-dopa were measured in two different days, before cabergoline was added, and at the end of the study. In both studies motor performance was evaluated by means of the Unified Parkinson's Disease Rating Scale (motor examination) and the Clinical Global Impression Scale; on-off diaries of daily motor condition also were filled by fluctuating patients. In patients with de novo PD, cabergoline pharmacokinetic parameters were unmodified by the adjunct of L-dopa, except that the time to reach the peak concentration (Tmax) significantly increased after L-dopa. In patients with fluctuating PD, no modification of L dopa pharmacokinetics was observed before and after cabergoline coadministration. Clinical evaluations confirmed that cabergoline is effective in the treatment of advanced PD as well as in the management of de novo patients. PMID- 9331532 TI - Newer drugs in the treatment of idiopathic pulmonary fibrosis. PMID- 9331534 TI - Genetic predisposition of idiopathic pulmonary fibrosis. AB - The pulmonary parenchyma in interstitial fibrosis responds stereotypically to a variety of insults. The observation that only a small subset of persons exposed to known fibrogenic agents develop fibrosis and the induction of pulmonary fibrosis in genetically susceptible mice support a genetic predisposition. The description of familial idiopathic pulmonary fibrosis is the strongest evidence available supporting this hypothesis. Unfortunately, despite all the recent advances in molecular genetic techniques there have been few human studies to date. Given the poor prognosis and the lack of a cure for pulmonary fibrosis, future genetic studies will, it is hoped, clarify the pathogenesis of pulmonary fibrosis and lead to preventive measures and new therapeutic interventions. Idiopathic pulmonary fibrosis typically occurs in patients who are older than 50. The etiology is unknown. PMID- 9331533 TI - Airway obstruction in interstitial lung disease. AB - There is no question that most interstitial lung diseases result in structural alteration of the small airways as well as the alveoli. These structural changes of the airways produce airflow abnormalities that, depending on their extent and severity, are reflected in a variety of tests of pulmonary function. However, in most situations, obstructive lung disease rarely dominates the clinical picture. Airflow limitation may be the predominant defect in patients with Wegener's granulomatosis, lymphangioleiomyomatosis, and chronic eosinophilic pneumonia. Concomitant risk factors such as cigarette smoking or dust inhalation may contribute to airway obstruction. Sporadic cases of interstitial lung disease progressing to overt airflow obstruction have been reported. The clinical significance of airway narrowing in interstitial lung disease is a maldistribution of ventilation that causes abnormalities on gas exchange, thereby increasing the work of breathing and possibly the sensation of dyspnea. PMID- 9331535 TI - Role of electron microscopy in interstitial lung disease. AB - Although electron microscopy no longer enjoys the important role in the diagnosis of interstitial lung diseases that it had in the 1960s and 1970s, it remains an important adjunct in the differential diagnosis of certain pulmonary diseases. Examples include various manifestations of systemic lupus erythematous pneumonitis, in which the presence of tubuloreticular structures and electron dense deposits are useful for diagnosis; immotile cilia disorders, in which qualitative and now quantitative studies of the cilia of respiratory epithelial cells can help to establish the diagnosis; infections by viruses and other subcellular microorganisms as shown by the role played by electron microscopy in the initial diagnosis of the Hantavirus pulmonary syndrome; pneumoconioses, in which, in conjunction with elemental analysis probes, scanning electron microscopy is of critical importance in establishing the presence of offending foreign compounds in lung tissue or fluids; pulmonary fibrilloses, such as amyloidosis, light chain disease, and fibrillary glomerulonephritis, affecting the lung; and cases of alveolar proteinosis or Langerhans cell granulomatosis diagnosed from fluids such as bronchoalveolar lavages or small tissue samples. As important, electron microscopy remains of enormous usefulness in the study of early structural events leading to the pathogenesis of diseases. For example, recent uses of the technique have focused on the alveolar-capillary wall damage induced by alveolitis in hypersensitivity pneumonitis and sarcoidosis. In summary, electron microscopy remains a useful method in the study and diagnosis of some interstitial lung diseases, but because of its expense it is incumbent on the clinician to use good judgment in the selection of cases and diseases for study by this method. PMID- 9331536 TI - Immune effector cells in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a poorly understood immunomediated disorder that is characterized by three major features; influx of inflammatory cells into the lower respiratory tract, alveolar epithelial or capillary cell injury, and release of cytokines that stimulate proliferation of fibroblasts and type II pneumocytes along with deposition of extracellular matrix proteins, notably collagen. In the past few years interest in the pathogenetic mechanisms taking place in IPF has focused on immune effector cells that are involved in pulmonary inflammatory pathways, cell-specific injury, and fibroblast activation. In particular, accumulating evidence indicates that a complex relationship exists between the macrophage/lymphocyte/neutrophil cellular axis and the local network of cytokines that, through paracrine and autocrine interactions, coordinate inflammation and fibrogenesis in the respiratory tract. PMID- 9331537 TI - Immunopathology of collagen vascular disease. AB - In this review, we focus on the heterogeneity of interstitial lung diseases detected in patients with collagen vascular diseases. By recognizing the heterogeneity of histopathology and comparing them with bronchoalveolar lavage fluid cell findings, we can understand profiles of lung inflammation and injuries and fibrosis in collagen vascular diseases. We focus on the significance of lung lymphocytosis in the lesions of patients with collagen vascular diseases, looking most closely at lesions in unusual interstitial pneumonia. The current understanding of immunopathogenesis and immunopathological findings is reviewed in the context of subsets of collagen vascular diseases. PMID- 9331538 TI - Pulmonary schistosomiasis. AB - Schistosomiasis is a very important infectious disease, and pulmonary involvement is not very rare. There may be two forms of pulmonary involvement, acute and chronic. The acute form usually occurs about 6 weeks after the infection (Katayama syndrome) and seems to be due to an allergic manifestation to the presence of Schistosoma spp. worm or eggs. The chronic form is more commonly seen in endemic areas and may cause pulmonary hypertension and cor pulmonale, pulmonary granulomatous schistosomiasis, and pulmonary arteriovenous fistulas. Recurrence of pulmonary infiltrations may appear after treatment. The globalization of the world with international travel makes it necessary for clinicians around the world to be aware of some "old" diseases from endemic areas of the globe. PMID- 9331539 TI - Newer therapeutic interventions for pulmonary transplant rejection. AB - Rejection of the lung allograft remains a significant problem and the primary cause of morbidity and mortality for the transplant recipient. Various strategies have been developed to prevent and minimize episodes of rejection. These methods involve either specific inhibitors or using a combination of barriers at precise sites in the immune response to induce graft tolerance. Another technique would be to create a tranquil environment between opposing populations of immunocompetent cells from both the donor and recipient by enhancing chimerism. Finding potent immunosuppressive agents is not the only impediment to prolonged graft survival. The challenge is also to develop techniques and agents that do not have global immunosuppressive properties that would cause the host to become more susceptible to infectious agents, as well as to prevent toxicity to other vital organs. A review of available new pharmaceutical therapies and an overview of potential future methods are presented here. PMID- 9331540 TI - Is nuclear imaging of any value in managing interstitial fibrosis? AB - The author describes a current status of nuclear imaging in interstitial lung disease. A number of tests have been proposed for the evaluation of disease activity. The only one that is used in many centers, however, is 67Ga scan. If the goal is the evaluation of functional impairment, 99mTc-penta-acetic acid aerosol may be a good tool to measure alveolar-capillary membrane permeability, but the test is not very popular because it is time consuming and may scatter radioactivity in the air. Perfusion imaging, which is more simple, reflects the vascular component of pulmonary parenchymal involvement more accurately than the chest radiograph and in one study has shown changes that could not be seen on the chest radiograph in 26% of cases. PMID- 9331541 TI - Incidence and recognition of interstitial pulmonary fibrosis in developing countries. AB - Interstitial pulmonary fibrosis in developing countries is now diagnosed with an increased frequency. Increased awareness and more frequent availability of computed tomography and fiberoptic bronchoendoscopy have helped in making the diagnosis more often. The spectrum of diseases causing pulmonary fibrosis is broadly similar to that seen in the West. Connective tissue disorders such as systemic sclerosis and rheumatoid arthritis and sarcoidosis are more common causes. Idiopathic fibrosis is seen in approximately half the patients. Pneumoconiosis such as silicosis are also important. Diagnosis is often established on the basis of clinical features and radiologic findings alone. Transbronchial lung biopsy is used as a frequent method to make histologic diagnosis. Some of the causes described from India are rather rare. One of the interesting examples included a patient in whom pulmonary fibrosis was related to his ascent to very high altitude. Extreme cold, solar radiation, and other factors complicating low atmospheric oxygen pressure were implicated as causative factors. Lung fibrosis, secondary to exposure to toxic gas (methyl isocyanate), is reported in survivors of the Bhopal gas leakage tragedy of 1984. Serial bronchoalveolar studies have show elevated fibronectin levels and the presence of macrophage-neutrophilic exudate in the lavage fluid. PMID- 9331542 TI - Pulmonary involvement in unusual multisystem disorders. AB - The clinical syndrome of interstitial lung disease encompasses a number of clinical disorders that affect the alveolar walls, interstitium, and vicinal structures including small airway and pulmonary vasculature. The syndrome has more than 200 causes including bacterial, viral, fungal, protozoal, and parasitic infections; collagen vascular disorders including systemic lupus erythematosus, rheumatoid arthritis, progressive systemic sclerosis, ankylosing spondylitis, mixed connective tissue disease, Sjogren's syndrome, and various vasculitides; hypersensitivity lung disease or extrinsic allergic alveolitis; inorganic neumoconioses including silicosis, asbestosis, and berylliosis; drug-induced and iatrogenic entities; and disorders of unknown origin including sarcoidosis, idiopathic pulmonary fibrosis, eosinophilic granulomatosis, lymphangioleiomyomatosis, and bronchiolitis obliterans-organizing pneumonitis. Apart from these, there are many uncommon metabolic and immune disorders that can involve pulmonary interstitium. Recently developed epidemiologic, immunologic, and molecular biology techniques undoubtedly will add to the list other entities that may produce interstitial lung disease. PMID- 9331543 TI - Hypersensitivity pneumonitis. AB - There are 30 or more groups of hypersensitivity pneumonitis (HP), such as farmer's lung, bird fancier's disease, humidifier lung, air-conditioner disease, and summer-type HP. Regardless of the causative agent or its environmental setting, the pathogenesis and clinical manifestations of the groups are similar. Immune-complex formation and complement activation might play a role during the early inflammatory phase of the disease. Much evidence, however, supports a more important role of T-cell-mediated delayed-type hypersensitivity reaction than humoral hyperresponsiveness in the development of the disease. High-resolution CT findings, a striking increase in the number of T cells in bronchoalveolar lavage fluids, and the presence of specific IgG and IgA antibodies to the causative antigens in the patient's serum samples are helpful in differentiating HP from other interstitial lung diseases. Management and treatment involve avoidance of antigen exposure and occasional use of corticosteroid therapy. PMID- 9331544 TI - Interstitial lung disease. PMID- 9331545 TI - Peptidomimetic regulation of growth hormone secretion. PMID- 9331546 TI - Catch-up growth. PMID- 9331547 TI - Molecular genetics of human hypertension: role of angiotensinogen. PMID- 9331548 TI - Excitatory amino acids: evidence for a role in the control of reproduction and anterior pituitary hormone secretion. PMID- 9331549 TI - Breast cancer and the role of cytokines in regulating estrogen synthesis: an emerging hypothesis. PMID- 9331550 TI - Roles of circadian rhythmicity and sleep in human glucose regulation. PMID- 9331551 TI - The Wellcome Prize Lecture. Calcium entry mechanisms in human platelets. PMID- 9331552 TI - Nitric oxide in the control of submandibular gland function in the anaesthetized ferret. AB - Stimulation of parasympathetic innervation of the submandibular gland (2 or 20 Hz continuously or 20 Hz for 1 s at 10 s intervals), in the ferret, produced secretion of fluid and protein and a fall in vascular resistance. The responses following the administration of N omega-nitro-L-arginine methyl ester (L-NAME; 2 mmol kg-1 i.a.) to block the synthesis of nitric oxide (NO) were reduced, and the persisting responses were abolished (at 2 Hz continuously and 20 Hz intermittently) or further reduced (at 20 Hz continuously) by the additional administration of atropine. The output of vasoactive intestinal peptide (VIP) from the gland was not affected. Neither the secretory nor the vascular response to intra-arterial infusions of acetylcholine (1.25 nmol kg-1) was affected by L NAME, whereas the vascular responses to both VIP (10 pmol kg-1) and pituitary adenylate cyclase-activating peptide (1-38) (PACAP) (0.5 pmol kg-1) were reduced thereby. Neither peptide evoked a fluid secretion per se. However, when infused during parasympathetic stimulation of saliva secretion, VIP increased both flow rate and the output of protein. These effects of VIP were abolished by L-NAME. The experiments were performed in the presence of sodium nitroprusside at doses (4-8 nmol min-1 kg-1 i.v.) aimed to counterbalance the systemic effects of L NAME. The results show that, in the ferret, parasympathetic nerve activity increases submandibular blood flow, and elicits the flow of saliva and output of protein by mechanisms that involve in situ generation of NO, upon which the effects of VIP and PACAP but not acetylcholine are largely dependent. PMID- 9331553 TI - Reversal of fusimotor reflex responses during locomotion in the decerebrate cat. AB - The effect of brief trains of electrical stimulation, at 2, 3 and 20 x threshold (T), of cutaneous afferents in the medial plantar nerve on the discharges of single medial gastrocnemius static and dynamic gamma-efferents has been investigated at rest and during locomotion in a decerebrate cat preparation. The units were classified as dynamic (10 units) or static (10 units) indirectly on the basis of their resting and locomotor discharge characteristics. Responses were assessed by calculating the change in mean gamma-rate during the 100 ms after stimulus onset compared with a control period. At rest, most dynamic neurones were inhibited by stimulation at 2T (9 of 10 units) and above. In contrast, the resting responses of most static neurones were excitatory at 2T (9 of 10 units) and 3T, while 20T produced static gamma-effects that varied in sign. During locomotion the responses of both types of gamma-efferent were phase related. Two patterns were observed with dynamic units. For seven dynamic neurones, at stimulus levels of 2T (7 units) and above, responses during electromyogram (EMG) bursts were inhibitory while those between bursts were not significantly different from zero. However, for three other dynamic units, a phase-related reversal of reflex responses was observed at some stimulus intensities (always 2T, 3 units) comprising inhibition during, and excitation between, EMG bursts. For static neurones, inhibitory (never excitatory) responses occurred during walking at stimulus intensities of 2T (10 units) and above. The locomotor responses of static units were maximum during (3 units) or between (7 units) EMG bursts and were minimum in the opposite phase of EMG activity. A task related reversal of reflex responses was thus generally apparent (9 of 10 units) to low intensity stimulation (2T) for static gamma-efferents during locomotion (inhibition) compared with rest (excitation). During locomotion there was a significant linear relation between the magnitude of response and the background gamma-rate for static units and those dynamic units that did not exhibit phase related reflex reversal (total, 17 units). For dynamic gamma-efferents, inhibition at rest and during locomotion occurred at short (spinal) latencies which were not significantly different and are consistent with the involvement of the same interneuronal pathway. We conclude that pathways of opposite sign may dominate the responses of fusimotor neurones to low threshold cutaneous afferents from the plantar surface of the foot depending on behavioural context. Furthermore, the cutaneous reflex responses of both types of gamma-motoneurones during locomotion appear to vary with the source of the afferent input and do not constitute a general excitatory drive. The results are discussed in relation to the role and reflex control of the fusimotor system. PMID- 9331554 TI - Electrophysiological characteristics of submucosal neurones in the proximal colon of guinea-pigs: comparisons with caecum and descending colon. AB - A systematic examination has been made of the active and passive electrophysiological properties and synaptic inputs of forty-four randomly impaled submucosal neurones in the proximal colon of the guinea-pig to compare these characteristics directly with those of submucosal neurones in the caecum (n = 70) and descending colon (n = 45). Within each of the three electrophysiological classes of submucosal neurones identified (S, S/AH and AH), no statistically significant regional differences were found with respect to the resting membrane potential, membrane time constant or input resistance between neurones of the proximal colon, descending colon and caecum. Of submucosal neurones from the proximal colon, forty-three of forty-four (98%) received fast excitatory synaptic potentials (fast EPSPs); thirty-nine (91%) were S neurones and the others were S/AH neurones; only one of the forty-four cells (2%) was an AH neurone. An idazoxan-sensitive slow inhibitory postsynaptic potential (slow IPSP) was induced in thirty of forty-three S and S/AH neurones (70%) of the proximal colon, compared with sixty-one of sixty-six caecal neurones (92%) and twelve of forty-one neurones (29%) in the descending colon. The mean (+/- S.E.M.) amplitude of the slow IPSP in proximal colonic neurones was 17 +/- 1 mV (range, 6 30 mV; n = 30), compared with the significantly larger synaptic response (25 +/- 1 mV; range, 7-38 mV; n = 66; P < 0.05) recorded in the caecum; the mean slow IPSP amplitude in the descending colon was significantly smaller (12 +/- 2 mV; range, 5-27 mV; n = 12; P < 0.05) than that in the caecum. In the proximal colon and caecum, only those neurones with a slow IPSP had a hyperpolarizing response to noradrenaline, whereas about 50% of those neurons of the descending colon that lacked a slow IPSP were hyperpolarized by noradrenaline, acting via alpha 2 adrenoceptors. Thus, the electrophysiological characteristics of the submucosal neurones of the proximal colon more closely resemble those of the caecum than those of the descending colon, of which many do not have a functional noradrenergic synaptic input. Furthermore, the results confirm that there are fundamental regional differences in the guinea-pig large intestine with respect to the synaptic organization of submucosal neurones of particular electrophysiological classes. PMID- 9331555 TI - Pituitary adenylyl cyclase-activating polypeptides and vasoactive intestinal peptide inhibit bone resorption by isolated rabbit osteoclasts. AB - Nerve fibres present in the periosteum, cortical bone and bone marrow are proposed to be involved in the regulation of bone metabolism by the release of neuropeptides acting locally on bone cells. The present study describes the effects of vasoactive intestinal polypeptide (VIP), shown to be present in the vicinity of bone, and the two related neuropeptides pituitary adenylyl cyclase activating polypeptide(1-38) (PACAP-38) and PACAP-27 on bone resorption in vitro induced by osteoclasts isolated from 10-day-old rabbits. Bone resorption was measured as the number and total area of pits formed by tartrate-resistant acid phosphatase-positive multinucleated cells (TRAP + MNCs) cultured for 3 days on devitalized slices of bovine cortical bone. All three neuropeptides had significant inhibitory effects on both the number and area of pits formed. At a high concentration (10(-7) M) the mean +/- S.E.M. reductions in the total area resorbed compared with controls were 70.3 +/- 8.2, 45.2 +/- 7.3 and 63.4 +/- 7.2% for PACAP-38, PACAP-27 and VIP, respectively. The corresponding values for the apparent dissociation constants were 0.93 +/- 0.30, 3.2 +/- 1.6 and 0.35 +/- 0.14 nM, respectively. The number of TRAP + MNCs was unaffected by application of neuropeptides. Autoradiography showed the presence of 125I-VIP binding sites on some stromal cells, whereas osteoclasts had no binding sites for 125I-VIP. A high number of 125I-calcitonin binding sites was demonstrated on osteoclasts in the same preparation. The signal transduction pathway remains to be fully elucidated but seems to be partly dependent on changes in intracellular calcium concentrations, since a number of stromal cells responded to application of 10( 8) M PACAP-38 or VIP, and at least partly independent of cAMP accumulation. Trifluoperazine and mellitin, two selective calmodulin inhibitors, failed to block the VIP-induced inhibition of bone resorption. Our results demonstrate a non-toxic and probably stromal cell-derived effect of PACAP-38, PACAP-27 and VIP on isolated rabbit osteoclasts, resulting in a potent inhibition of bone resorption in vitro. The signal transduction pathway for inhibition induced by PACAP-38, PACAP-27 and VIP may be mediated partly by changes in intracellular Ca2+ in stromal cells. PMID- 9331556 TI - A quantitative description of dynamic left ventricular geometry in anaesthetized rats using magnetic resonance imaging. AB - We report a functional application of magnetic resonance imaging (MRI) for the quantitative description of left ventricular geometry through systole and diastole in normal anaesthetized Wistar rats that might be applicable for the analysis of chronic changes resulting from pathological conditions. Images of cardiac anatomy were acquired through planes both parallel and perpendicular to the principal cardiac axis at times that were synchronized to the R wave of the electrocardiogram. The images of the transverse sections were assembled into three-dimensional representations of left ventricular geometry at consecutive time points through the cardiac cycle. This confirmed the geometrical coherence of the data sets, that each slice showed circular symmetry, and that the images were correctly aligned with the appropriate anatomical axes. Different models for the three-dimensional geometry of the left ventricle were then tested against the epi- and endocardial surfaces reconstructed from images of the transverse sections of the left ventricle in both systole and diastole using least-squares minimizations in three dimensions. In agreement with previous reports in the human heart, an elliptical figure of revolution offered an optimal fit to the epicardial and endocardial geometry for the rat heart in diastole. This was in preference to models that used spherical, quartic or parabolic geometries. However, in contrast to contraction in the human heart, all these geometrical representations broke down during systolic ejection in the rat heart. We therefore introduced a more general hybrid model which described left ventricular geometry in terms of the variation of the radii r(z), independently determined for each slice, with its position z along the principal cardiac axis. The resulting function r(z) could then be described by a simple ellipsoid of revolution not only during diastole, but also throughout ventricular ejection. The findings also ruled out alternative geometrical representations. It was then possible additionally to reconstruct the luminal and total left ventricular volumes, wall thicknesses and ejection fractions through the cardiac cycle and to confirm that the predicted total ventricular wall volume was conserved throughout the cardiac cycle. Our hybrid model of cardiac geometry may thus be useful for non-invasive serial studies of chronic pathological changes that use the rat as a model experimental system. PMID- 9331557 TI - Parotid secretion daily patterns and measurement with ultrasonic flow probes in conscious sheep. AB - Five sheep under halothane anesthesia were prepared with bilateral transit time ultrasonic flow probes around the parotid ducts. The ducts were fitted with non obstructive sampling catheters through their oral ends. After probe encapsulation (8 days), salivary flows were continuously recorded (4-5 days, dual-channel ultrasonic flowmeter). For rumination, eating, resting and drinking periods, respectively, the parotid daily outputs recorded were 1.96 +/- 0.57, 0.97 +/- 0.34, 2.84 +/- 0.41 and < 0.041 and bilateral flow rates were 6.76 +/- 0.70, 5.63 +/- 1.42, 2.50 +/- 0.58 and 1.69 +/- 0.88 ml min-1. An ipsilateral secretory reflex was evident when the sheep changed chewing side during rumination (4.44 +/ 0.96 ml min-1 ipsilateral vs 2.63 +/- 0.90 ml min-1 contralateral flow, P < 0.01). Secretory patterns are described in detail during rest, eating, drinking and rumination periods. The pH of parotid saliva (8.36 +/- 0.14) and the osmolality (273.8 +/- 9.9 mosmol kg-1) were independent of secretory rates. In situ probe calibration showed high accuracy (0-9%). The main advantages of the technique are its accuracy and good tolerance, duct integrity and maintenance of nervous supply, minimal surgery, uninterrupted salivary flow, simultaneous bilateral measurements and precise flow monitoring, permitting detailed observations. PMID- 9331558 TI - Mechanisms responsible for changes in abdominal vascular volume during sympathetic nerve stimulation in anaesthetized dogs. AB - This study was designed to determine the extent to which the decrease in volume of blood in the abdominal circulation in response to sympathetic stimulation was due to a passive effect of decreasing flow rather than active constriction of the capacitance vessels. In dogs anaesthetized with alpha-chloralose (100 mg kg-1 i.v.) the abdominal circulation was vascularly isolated and perfused either at constant flow or at constant pressure, and drained at constant pressure from the inferior vena cava. Changes in volume were determined by integration of the differences between inflow and outflow. Supramaximal stimulation of both splanchnic (sympathetic) nerves at 1 Hz decreased abdominal volume during constant pressure perfusion (active and passive components) by 3.04 +/- 0.58 ml kg-1 and at constant flow (active responses only) by 2.30 +/- 0.49 ml kg-1 (means +/- S.E.M.). The responses at 8 Hz were respectively 9.52 +/- 0.91 and 5.09 +/- 0.49 ml kg-1. The proportion of the responses calculated to be passive at 1 and 8 Hz was 23 +/- 6.3 and 45 +/- 5.1%, respectively. These responses were almost identical to those induced by changing inflow by increasing the pump speed. Following ligation of the splenic pedicle, the responses during both constant pressure and constant flow were reduced by similar amounts, indicating that only the active response was affected. After ligation of the splenic pedicle, the proportion of the response calculated to be passive at 1 and 8 Hz increased to 44 +/- 8.0 and 62 +/- 3.7% respectively. These results indicate the importance of passive volume change in affecting abdominal volume, particularly following ligation of the splenic circulation. PMID- 9331559 TI - Glomerular haemodynamics during renal artery clamping and haemorrhage in the dog. AB - The influence of gradual decline in renal perfusion pressure (RPP) due either to renal artery clamping (C) or to haemorrhagic hypotension (HH) was studied using micropuncture techniques in anaesthetized dogs. The decrease in renal blood flow (RBF) was more profound and set in earlier during HH than during C, where perfect autoregulation was observed down to a mean arterial blood pressure of 85 mmHg. Glomerular filtration rate (GFR) was also only slightly decreased during C, with no change in filtration fraction (FF); again, a much greater decrease in GFR with an increase in FF was seen in HH. The excretion of water, electrolytes and urea were also more decreased during HH than during C. Similar changes were seen at the single nephron (SN) level. Opposite changes were observed in arteriolar resistances: during C a decrease in total arteriolar resistance (RT) amounting to -22% at a RPP of 84 mmHg and -13% at 60 mmHg was seen, due exclusively to a drop in afferent resistance (RA), but during HH there was a significant increase in RT by +36% at RPP of 110 mmHg, +39% at 85 mmHg and +68% at 60 mmHg. This increase was mainly due to an increase in efferent resistance (RE) rather than in RA: +42 vs. +31%, respectively, at 110 mmHg and +67 vs +19% respectively, at 85 mmHg. It was not until a RPP of 60 mmHg was reached that this difference between RE and RA disappeared, being +67% for RE and +69% for RA. The ultrafiltration coefficient, Kf, did not change during C and only decreased slightly with the biggest drop in RPP during HH (2.84 microliters mmHg-1 min-1 during HH vs. 4.19 microliters mmHg 1 min-1 before HH). The SNGFR/GFR ratio remained unchanged during C but declined with decreasing RPP during HH, which probably indicates a 'redistribution' of RBF to the deeper regions of the renal cortex. In conclusion, major differences in renal function were observed between C and HH whose cause is unknown. PMID- 9331560 TI - Programmed cell death in bovine mammary tissue during lactation and involution. AB - Cessation of milk removal causes mammary tissue involution, which in rodents is characterized by extensive tissue degeneration and loss of the majority of luminal epithelial cells by apoptosis. In contrast, bovine mammary tissue shows little histological evidence of tissue remodelling between lactations. In this study, we combined histology with molecular biology to examine the cellular and molecular changes in bovine mammary tissue on cessation of milking. Oligonucleosomal laddering of genomic DNA extracted from lactating tissue indicated that a proportion of cells were dying by apoptosis. This was confirmed by terminal deoxynucleotide transferase-mediated deoxyuridine nick end-labelling of apoptotic cells in lactating tissue sections (TUNEL). One week after cessation of milking, alpha-lactalbumin and alpha S1-casein messenger RNA (mRNA) abundance had decreased by 99 and 85%, respectively, whereas lactoferrin mRNA had increased 20-fold. Drying off was also accompanied by an increase in oligonucleosomal laddering of genomic DNA, and by an increase in the proportion of TUNEL-positive cells, which were localized preferentially in regions where alveolar structure had deteriorated. Therefore, termination of lactation was associated with partial loss of the mammary cell population and dedifferentiation of the remainder. PMID- 9331561 TI - Identification of additional complementation groups that regulate genomic instability. AB - By somatic cell hybridization, amplification has been found to be a recessive genetic trait in three tumor cell lines examined. Studies with transgenic mice have shown that amplification frequency can be altered by a lack of wild-type TP53 (p53) activity. Other factors may regulate this phenotype in tumor cell lines possessing both wild-type p53 activity and amplification ability. Complementation analysis of somatic cell hybrids was performed to delineate groups of tumor cell lines that share a common defect that modulates the ability to amplify. The amplification frequencies of three normal fibroblast x tumor hybrids were suppressed 10-100-fold from parental tumor values, extending the observation that amplification is a recessive genetic characteristic in these cell lines. Analysis of tumor x tumor hybrids revealed at least two complementation groups. Defects in these groups differed from TP53 and implicate multiple variables in the regulation of gene amplification. PMID- 9331562 TI - Philadelphia-like translocation t(9;22)(q34;q11) found in a follicular lymphoma involving not BCR and ABL but IGL-mediated rearrangement of an unknown gene on 9q34. AB - In a case of follicular center cell lymphoma (FCCL) without evidence of histologic progression towards a high-grade lymphoma, t(9;22)(q34;q11) was found simultaneously with a t(14;18)(q32;q21) and a t(8;14)(q24;q32). Molecular studies of this case showed BCL2 and MYC rearrangements in addition to the rearrangements of immunoglobulin heavy (IGH) and lambda (IGL) loci. Investigation of the t(9;22) using Southern blot and RT-PCR analysis failed to detect M-bcr or m-bcr rearrangements of BCR. Two-color fluorescence in situ hybridization (FISH) with ABL and BCR probes revealed presence of a "fusion" signal, but its atypical localization [der(9)] and gene order [cen-ABL-BCR-tel] indicated that this translocation differed from the t(9;22) in chronic myeloid leukemia and did not involve either ABL or BCR. In addition, further FISH analysis using 9q34- and 22q11-specific probes localized the breakpoint on chromosome 9 distal to the NOTCH1 gene and the breakpoint on 22q11 in the IGL gene cluster. These results indicate an IGL-mediated rearrangement of an unknown gene at 9q34 that together with BCL2 and MYC might be involved in the lymphomagenesis of the present case of FCCL and perhaps in other cases of non-Hodgkin lymphoma in which t(9;22) is sporadically occurring. PMID- 9331563 TI - Cytogenetic analysis of three breast carcinoma cell lines using reverse chromosome painting. AB - Chromosome painting was used to determine the copy number and identity of virtually all the chromosomes in three breast cancer cell lines, T-47D, MDA-MB 361, and ZR-75-1. The karyotypes of all three cell lines were very complex, and were consistent with the monosomic pattern of evolution suggested by Dutrillaux, in which nonreciprocal translocations cause an initial reduction in chromosome number, followed by duplication of the entire genome and further chromosome loss. Twenty distinct abnormal chromosomes were identified in T-47D, seven of which were present as two copies. MDA-MB-361 had 27 abnormal chromosomes each as a single copy. Thirteen abnormal chromosomes in ZR-75-1 occurred singly, two were paired, and one was present as three copies. Most of the aberrant chromosomes were nonreciprocal translocations, although deletions, duplications, isochromosomes, and amplifications (HSR of 1q) were also found. Chromosome arms present in abnormal chromosomes in all three lines were 1q, 6p, 7p, 8p, 8q, 10q, 11p, 11q, 12p, 13q, 14q, 15q, 16p, 16q, 17q, and 20q. The only chromosome arms present in four or more copies in all three lines were 8q and proximal 12p, while 1p, 17p, and bands 11q12--13 were the only chromosome regions consistently reduced to two copies. The most striking feature common to all three lines was a translocation breakpoint on the short arm of chromosome 8 at 8p12. PMID- 9331564 TI - Allelic losses on 18q21 are associated with progression and metastasis in human prostate cancer. AB - We analyzed normal/tumor DNA pairs obtained from 46 patients with prostate cancers (stage B, 16 cases; C, 10 cases; D1, 4 cases; and endocrine therapy resistant cancer-death, 16 cases) for loss of heterozygosity using 32 microsatellite markers on chromosome 18. Seventeen of the 46 cases (37%) showed loss of heterozygosity (LOH) for at least one locus on the long arm. Detailed deletion mapping in these tumors identified a distinct commonly deleted region within a 5-cM interval in 18q21.1. There was a statistical correlation between the frequency of LOH on 18q and clinical stage (chi 2 = 12.3; P = 0.0064). LOH on 18q was observed more frequently in Stage D1 cases (4/4; 100%) than in stage B+C cases (5/26; 19%; P = 0.0046, Fisher's exact test). In 8 of 9 (89%) cancer-death patients from whom DNAs were available from both primary and metastatic tumors, the primary tumors had either no detectable abnormality of chromosome 18 or the region involving loss of heterozygosity was limited while the metastatic foci showed more frequent and extended allelic losses on this chromosome. No abnormalities were detected in the DCC and DPC4 genes when their exons were analyzed separately by single strand conformation polymorphism assay. These results suggest that inactivation of one or more putative tumor suppressor genes on 18q21 other than DCC and DPC4 plays an important role in the progression of human prostate cancer. PMID- 9331565 TI - Location of the BCR-ABL fusion gene on the 9q34 band in two cases of Ph-positive chronic myeloid leukemia. AB - Two new variant Philadelphia (Ph) chromosomes with an aberrant location of the BCR-ABL fusion gene on 9q34 of the derivative 9 are reported. One presented cytogenetically as a standard t(9;22)(q34;q11), whereas the other was classified as an ins(9;22)(q34;q11.1q11.2) using the combined interpretation of cytogenic, FISH, and molecular data. The mechanisms of the two rearrangements are presented. It is suggested that the insertion has occurred in a single event in the patient with ins(9;22). In the patient with t(9;22), both a translocation and an insertion, occurring either sequentially or simultaneously, can account for the location of the BCR-ABL fusion gene on the derivative 9. A possible poor prognostic impact of this aberrant location of the BCR-ABL is also suggested by the clinical data reported in such patients. PMID- 9331566 TI - FISH identifies different types of duplications with 12q13-15 as the commonly involved segment in B-cell lymphoproliferative malignancies characterized by partial trisomy 12. AB - Clinical, cytogenetic, fluorescence in situ hybridization (FISH), and Southern blot data of 18 patients with different subtypes of B-cell non-Hodgkin's lymphoma, cytogenetically characterized by partial trisomy 12, are presented. These chromosomal changes occurred predominantly in clinically progressive chronic lymphocytic leukemia, mixed cell type, and advanced-stage follicle center cell lymphoma at the time of relapse or transformation into diffuse large cell lymphoma. Partial trisomy 12 consistently included the long arm of chromosome 12, either completely or partially, and resulted from dup(12q) or other rearrangements involving chromosome 12. The duplications were cytogenetically identified as dup(12)(q13q23), dup(12)(q13q22), or dup(12)(q13q15) in follicle center cell lymphoma or t(14;18)-positive diffuse large cell lymphoma; dup(12)(q13q22) or dup(12)(q13q24) in chronic lymphocytic leukemia; and dup(12)(q13q21) in a case of t(14;18)-negative diffuse large cell lymphoma. FISH, using library probes and a panel of YAC probes, mapped along the long arm of chromosome 12, confirmed the cytogenetic results in all cases analyzed except for three cases of t(14;18)-positive follicle center lymphoma or diffuse large cell lymphoma with dup(12q). In these cases, FISH showed similar, possibly identical, duplications, which involved a region more centromeric (12q11-21) than assumed by karyotypic analysis (12q13-22 or 12q13-23) and included alphoid DNA sequences, a combination hitherto unknown. In addition, commonly duplicated regions of chromosome 12 could be defined: 12q11-21, including alphoid DNA sequences for follicle center cell lymphoma or t(14;18)-positive diffuse large cell lymphoma, 12q13-22 for chronic lymphocytic leukemia, and 12p13-q15 for marginal zone cell lymphoma, all of which overlapped in 12q13-15. Whether these regions, especially 12q13-15, may contain genes which are important in malignant transformation or disease progression of B-cell lymphoproliferative malignancies characterized by complete or partial trisomy 12 remains to be determined. PMID- 9331567 TI - Fine mapping of a region of common deletion on chromosome arm 10p in human glioma. AB - Allelic loss on chromosome 10 is a frequent event in high grade gliomas. Earlier studies have shown that in most cases a complete copy of chromosome 10 is lost in the tumor. To define more accurately and specifically the region of common deletion on chromosome arm 10p, we have screened a large series of gliomas for allelic losses that exclusively affect this part of the chromosome. Allelic loss profiles were determined for 127 gliomas, including 118 astrocytomas of various malignancy grades. Seventeen tumors displayed loss of part of chromosome 10. In three of these, only chromosome arm 10p sequences were lost. The interval between loci D10S559 and D10S435 in 10p15, with a length of approximately 800 kilobase pairs, was commonly deleted in the latter tumors, suggesting that this region may harbor a tumor suppressor gene important in glioma tumorigenesis. Comparison of the allelic loss profiles in the low and high grade astrocytomas revealed that astrocytoma progression is associated with increased loss of chromosome 10 sequences. PMID- 9331568 TI - Low level of DNA repair in human chromosome 1 heterochromatin. AB - Band 1q12 is a breakage prone region commonly involved in chromosome 1 rearrangements found in human cancer. We have investigated whether a lack of DNA repair can account for the observed 1q12 fragility. Ethyl methanesulfonate (EMS) induced repair sites originating in G0/G1 phase of the cell cycle were converted to chromosome breaks by blocking the re-synthesis step of excision repair with cytosine arabinoside. Thus, unfilled repair gaps reached S-phase and were detected as chromosome breaks after cell division. Chromosome breakage in the overall genome was monitored by scoring of micronuclei. Breaks in 1q12 were detected in interphase lymphocytes by in situ hybridization with tandem probes targeting the chromosome 1 centromere (D1Z5) and the adjacent heterochromatic band 1q12(pUC1.77). Mitomycin-C (0.12 and 0.24 micrograms/ml) and X-rays (1 and 2 Gy) were used as positive controls to check the suitability of the tandem labelling approach in detecting 1q12 clastogenicity. Our data indicated that, although the methodology was suitable to detect 1q12 breakage and a high level of overall genome DNA repair occurred, there was a low level of EMS-induced DNA repair sites in 1q12 converted to chromosome breaks. This finding was consistent throughout G1 phase, suggesting that there is a relatively low level of DNA excision repair in human chromosome 1 heterochromatin. PMID- 9331569 TI - Identification of complex genomic breakpoint junctions in the t(9;11) MLL-AF9 fusion gene in acute leukemia. AB - The MLL gene at chromosome 11, band q23, is involved in translocations with as many as 40 different chromosomal bands. Virtually all breakpoints occur within an 8.3 kb BamHI fragment and result in 5' MLL fused to partner genes in a 5'-3' orientation. The translocation t(9;11)(p22;q23), which results in the fusion of MLL to AF9, is the most common of the 11q23 chromosomal abnormalities observed in de novo acute myeloid leukemia (AML), in therapy related leukemia (t-AML), and rarely in acute lymphoblastic leukemia (ALL). We have studied 24 patients with a t(9;11) and an MLL rearrangement, including 19 patients with AML, four with t AML, and one with ALL. To understand the mechanisms of this illegitimate recombination, we cloned and sequenced the t(9;11) translocation breakpoint junctions on both derivative chromosomes from one AML patient and from the Mono Mac 6 (MM6) cell line, which was derived from a patient with AML. Two different complex junctions were noted. In the AML patient, both chromosome 11 and 9 breaks were staggered, occurred in Alu DNA sequences, and resulted in a 331 bp duplication. In the MM6 cell line, breaks in chromosomes 11 and 9 were also staggered, but, in contrast to the finding in the AML patient, the breaks did not involve Alu DNA sequences and resulted in a 664 bp deletion at the breakpoints. Using reverse transcriptase (RT-) PCR, we analyzed 11 patient samples, including the two just described, for MML-AF9 fusions. The fusion occurred in six of seven AML patients, two of two t-AML patients, one patient with ALL, and in the MM6 cell line. Interestingly, all of the breaks within the AF9 gene in AML patients occurred in the central AF9 exon, called Site A by others, whereas in the single ALL patient the breakpoint mapped to a more 3' region of the AF9 gene. Our data, when combined with those of others, suggest that the fusion point within the AF9 gene, and thus the amount of AF9 material included in the MLL-AF9 fusion gene product, may influence the phenotype of the resulting leukemia. This further supports the proposal that the MML translocation partner genes play a critical role in the leukemogenic process. PMID- 9331570 TI - Isolation of osteosarcoma-associated amplified DNA sequences using representational difference analysis. AB - Comparative genomic hybridization analysis of a primary osteosarcoma and its metastasis revealed two regions of DNA amplification, one at 17p11.2-12 and one at 19q12-13. Subsequent representational difference analysis of the primary tumor resulted in the isolation of two distinct tumor-amplified DNA fragments originating from chromosome 19. A YAC clone corresponding to one of the two isolated DNA fragments was used for fluorescence in situ hybridization on normal human lymphocyte metaphases and tumor-derived nuclei. This resulted in the localization of this YAC to 19q12-13.1 and confirmed the amplification status of the isolated fragment in the tumors. The availability of such RDA-isolated sequences may be instrumental in the search for genes relevant for tumor development. PMID- 9331571 TI - Breakpoints of 19q13 translocations of benign thyroid tumors map within a 400 kilobase region. AB - Structural rearrangements involving the long arm of chromosome 19 characterize a cytogenetic subgroup of benign thyroid tumors. To localize the breakpoint of the 19q13 aberrations, we have established three cell lines derived from benign thyroid tumors showing translocations in this region. We have used these cell lines and four additional primary tumors with 19q13 abnormalities for fluorescence in situ hybridization (FISH) mapping studies with ten cosmid clones located between the molecular markers POLD1 and TNNT1. The breakpoints of all chromosome 19 abnormalities mapped within a 400 kb region. PMID- 9331572 TI - Isolation, mapping and mutation analysis of a human cDNA homologous to the doc-1 gene of the Chinese hamster, a candidate tumor suppressor for oral cancer. AB - We have isolated a human cDNA encoding a 115-amino-acid polypeptide that revealed 97% identity to a candidate tumor suppressor gene for oral cancer in Mesocricetus auratus (deleted in oral cancer-1; doc-1). It also showed a high degree of homology to a gene induced by TNF-alpha in Mus musculus. To investigate its possible role in esophageal carcinogenesis, we examined genetic alterations and expression levels of the gene in 13 esophageal carcinoma cell lines and 10 primary esophageal carcinomas. No mutation nor reduction of expression was observed in any of the 23 cancer materials examined. These results imply that the human doc-1 homologue is unlikely to play a significant role in esophageal carcinogenesis, although its role in the TNF-alpha signaling pathway remains unclear. We mapped DOC1 to chromosome band 12q24.31 by fluorescence in situ hybridization. PMID- 9331573 TI - Combined chromosome microdissection and comparative genomic hybridization detect multiple sites of amplification DNA in a human lung carcinoma cell line. AB - Chromosome microdissection-fluorescence in situ hybridization and comparative genomic hybridization (CGH) were performed in parallel to identify the native location of amplified DNA in a human non-small cell lung cancer (NSCLC) cell line exhibiting a homogeneously staining region (hsr) and double minutes (dmin). The native locations of microdissected DNA from the hsr and dmin were 7p12-13 and 8q24, respectively. Southern analysis revealed coamplification of EGFR (7p12) and MYC (8q24). CGH detected amplification of DNA not only from 7p12-13 and 8q24, but also from 9p24 and 10q22. PMID- 9331574 TI - Biodisabilities in relation to other disease consequences in the functional assessment of patients with chronic low back pain. AB - The theoretical assumptions concerning the assessment of some physical disease consequences according to the Biopsychosocial Dimensional Classification (BPSDC) model were tested in patients with chronic low back pain. Factor analyses showed that the mutual relations of selected measures for biodisabilities, as well as their relationship to some measures of bioimpairments, and bio- and psycho handicaps supported many of the original assumptions of the BPSDC model. For instance, several dynamometer and non-dynamometer measures of physical capacity and performance associated themselves with the factor of 'physical fitness' interpreted to represent disabilities. A very interesting observation was that the subjective experiences of pain intensity, pain interference and management in various physical activities assessed by self-reports, represented the psycho-axis instead of the bio-axis. Another interesting notion was that the measures for organic pathology, i.e. 'neuromuscular findings' and 'spinal findings', separated themselves from other factors, and were interpreted to represent bio-impairments. Additionally, the range of motion measures in active movements and body positions were related to organic pathology, and were thus interpreted to bio-impairments, not disabilities as previously thought. It is emphasized that in order to know about the clinical meaningfulness of the dimensions found, further studies about the predictive value of the factors should be performed. PMID- 9331575 TI - The unemployed sick-listed and their vocational rehabilitation. AB - The unemployed on long-term sick-leave are many and difficult to rehabilitate. The likelihood that a period of sick leave will result in a temporary disability pension is about three times greater for the unemployed than for those with jobs. The aim of this current study was to examine the vocational rehabilitation process of the unemployed on long-term sick-leave. The study was based on 118 matched cases on long-term sick-leave, 59 employed and 59 unemployed. Data were collected at seven social insurance offices in rural areas of Jamtland, Sweden. Our hypothesis was that the unemployed were disregarded in vocational rehabilitation. The results partly confirm this. Unemployed people's potential need for rehabilitation is not investigated to the same extent as employed people's. Also the unemployed have to wait longer for an investigation. Against our hypothesis, however, no difference exists between the employed and the unemployed in rehabilitation impulse, rehabilitation plan or rehabilitation allowance. Nor do the unemployed, except for those awaiting investigation, wait longer than the employed. The major result of the study is, instead, that vocational rehabilitation in general, regardless of employment status, seems beset with problems. Rehabilitation activities seem far too few and initiated unnecessarily late. Neither the employer nor the social insurance office seem to be fulfilling their statutory duties. PMID- 9331576 TI - Vocational perspectives and neuromuscular disorders. AB - The present study analyses the actual occupational situation, vocational handicaps and past labour career of a group of about 1000 Dutch patients suffering from a neuromuscular disorder (NMD). On the basis of the likelihood of a substantial employment history and sufficient numbers of patients, four types of NMD were selected: dystrophia myotonica (DM), hereditary motor and sensory neuropathy, (HMSN), spinal muscular atrophy (SMA) and myasthenia gravis (MG). Results show that a labour career is in reach of most NMD patients, even for those with severe limitations. It is concluded that physical limitations seem not to be decisive in that respect. The loss of the quality of communication, the loss of mental abilities and the effect of the diseases on the facial expression, as with some DM patients, are also important for chances on the labour market. Though the labour participation of NMD patients tends to decrease after the age of 34, the availability of work adaptations makes it possible to prolong the labour career. Analysis of the actual work situation of NMD patients shows that both disorder-related limitations and work characteristics play an important role in the amount of physical work problems encountered. It is argued that physical labour has to be regarded as generally unsuitable for NMD patients. This has implications for the sort and level of education to be attained by NMD patients. Career counselling as a focus point for the choice of an educational programme may improve labour market opportunities as well as quality of employment of NMD patients. Allowing for and accepting the possible effects of the disorder in the work situation are considered to be important in respect to labour participation and work satisfaction of workers with NMD. Reducing time pressure demands and increasing the freedom to organize one's work, are measures to be given especial consideration. PMID- 9331577 TI - Descriptive epidemiology of physical activity in university graduates with locomotor disabilities. AB - The descriptive epidemiology of physical activity in a sample of 577 University of Illinois graduates (1952-1991) with locomotor disabilities was assessed by mail survey. The survey requested basic demographic information, age, gender, marital status, household income. Respondents were asked to rate their current activity levels and activity levels during their college years compared to others their age on a 5 point scale: (1) much less active to (5) much more active. Completed surveys were received from 229 alumni (40%); 59 semi-ambulatory, 115 paraplegic, 55 quadriplegic. Results indicated current physical activity was associated with mobility limitation. With more severe mobility limitations the percentage reporting being less/much less active increased (42.4% semi ambulatory, 56.5% paraplegic, 66.7% quadriplegic, P < 0.001) and the percentage reporting being more active decreased (20.3% semi-ambulatory, 16.5% paraplegic, 13.0% quadriplegic, P < 0.001). Current physical activity was significantly lower (P < 0.05) with increasing age, lower self-rated health, higher disability severity and among those who were sedentary during college. Physical activity did not differ by gender, marital status or household income. Multiple regression analysis indicated that health status was a significant predictor of current physical activity in all mobility categories (P < 0.001) after controlling for age, gender, income, disability severity and college activity. Among both paraplegics and quadriplegics physical activity during college was significantly associated (P < 0.001 paraplegic; P < 0.01 quadriplegic) with current physical activity. These results document a low level of physical activity in a well educated sample of individuals with locomotor disabilities and suggest that exposure to physical activity in an educational setting may be an effective technique for increasing physical activity in individuals with locomotor disabilities. PMID- 9331578 TI - The quality of life of people with mental retardation: in search of an adequate approach. AB - There is a lack of instruments that measure the quality of life of people with mental retardation. These types of instruments could be used in order to give an indication of the quality of care they receive. At the moment we are developing an instrument that measures quality of life. Our first task is to find an adequate definition of 'quality of life'. In this article an attempt is made to define this term as it relates to people with mental retardation. Starting from literature in the field of disabilities, reflections in the social sciences and philosophical analysis, a combined approach is adopted, according to which quality of life consists of specific objective and subjective factors. PMID- 9331579 TI - Needs of people with disabilities used to determine clinical education. AB - Western rehabilitation techniques were first introduced into China in the early 1980s. The predominantly 'medical model' of health care in China has resulted in a shortage of rehabilitation personnel and limited experience determining the needs of clients. Occupational therapy practitioners in China are learning their new roles and clients, having never had any contact with occupational therapy, are unaware what services can be offered. The purpose of this research was to determine the most frequently self-identified functional problems (in the ares of self-care productivity and leisure) identified by 113 in-patients in four different hospitals in China, in order to assist therapists to develop programmmes that are responsive to clients' needs. Using the Canadian Occupational Performance Measure, both males and females identified significantly more self-care than leisure or productivity problems (P = 0.01). Females identified more productivity problems than males (P = 0.01) while males appeared to consider leisure difficulties as a priority more frequently than women. The study was jointly undertaken by a Canadian and a Chinese researcher. To our knowledge, this is the first time that occupational therapy personnel in China have formally surveyed clients about their rehabilitation needs, in order to develop priorities for programming. PMID- 9331580 TI - Prediction of achieved mobility in prosthetic rehabilitation of the elderly using cognitive and psychomotor assessment. PMID- 9331581 TI - Assessing impact of disability awareness training using the Attitudes Towards Disabled Persons Scale (ATDP--form 0). PMID- 9331582 TI - Music therapy: facilitating behavioural and psychological change in people with stroke--a pilot study. PMID- 9331583 TI - Surveying the counselling needs of families with disablement in the United Arab Emirates. PMID- 9331584 TI - Socio-economic factors and childhood disability in Trivandrum of south India. PMID- 9331586 TI - Historical streptococci. PMID- 9331585 TI - Effect of a community-based computer skills training programme on self-concept changes of tutors with physical disabilities. PMID- 9331587 TI - A century of streptococcal research. PMID- 9331588 TI - Streptococcal research at Pasteur Institute from Louis Pasteur's time to date. PMID- 9331589 TI - The Streptococcus and the host. Present and future challenges. PMID- 9331591 TI - What is a throat culture? PMID- 9331590 TI - Rheumatogenic and nephritogenic group A streptococci. Myth or reality? An opening lecture. PMID- 9331593 TI - Antigen detection test for group A beta-hemolytic streptococcal pharyngitis that is sufficiently sensitive for use without confirmatory cultures. PMID- 9331592 TI - Group A streptococcal infections in hospitalized patients. PMID- 9331594 TI - Advantages of blood agar-NaCl selective medium in the isolation of beta-hemolytic streptococci from throat swabs. PMID- 9331595 TI - Opacity factor inhibition test for typing group A beta-haemolytic streptococcus strains and antibiotic susceptibility. PMID- 9331596 TI - Streptococcal pharyngitis in Argentina. A four-year study. PMID- 9331597 TI - Study of the streptococcal acute pharyngitis and carrier state. PMID- 9331598 TI - Streptococcus pyogenes bacteraemia. 107 episodes between 1970 and 1995. PMID- 9331599 TI - Invasive infections due to Streptococcus pyogenes in the Czech Republic. Results of the active surveillance. PMID- 9331600 TI - Detection of new DNA fragments integrated on the genome of M1 and M3 group A streptococci from streptococcal toxic shock-like syndrome. PMID- 9331601 TI - Two clusters of invasive Streptococcus pyogenes infection in England. PMID- 9331602 TI - Invasive group A streptococcal infections. Serotype newly associated with toxic shock-like syndrome in Trinidad. PMID- 9331603 TI - Invasive streptococcal disease (group A, B, and Streptococcus pneumoniae) in France 1987-1994. PMID- 9331604 TI - Clinical and microbiological characteristics of severe group A streptococcal infections in Italy. PMID- 9331605 TI - Comparison of invasive (septicemic) and non invasive strains of group A streptococci isolated during a one-year national survey in France. The Groupe d'Enquete 1995 sur les Infections Streptococciques. PMID- 9331606 TI - Necrotising fasciitis associated with invasive group A streptococcal infections in England and Wales. PMID- 9331607 TI - Characterisation of group A streptococci from necrotising fasciitis cases in Gloucestershire, United Kingdom. PMID- 9331608 TI - Role of Streptococcus pyogenes in the etiology of Erysipelas. PMID- 9331609 TI - Erysipelas--its occurrence and clinical aspects in Prague, 1993. A retrospective study. PMID- 9331610 TI - Searching for acute poststreptococcal glomerulonephritis-associated Streptococcus pyogenes in Australian aboriginal communities. PMID- 9331611 TI - Mitogenic factors from group G streptococci associated with scarlet fever and streptococcal toxic shock syndrome. PMID- 9331612 TI - Sydenham's chorea. Clinical and therapeutic update. PMID- 9331613 TI - Acute rheumatic fever in Tunisia. Serotypes of group A streptococci associated with rheumatic fever. PMID- 9331614 TI - Acute poststreptococcal glomerulonephritis in children. PMID- 9331615 TI - Poststreptococcal uveitis. PMID- 9331616 TI - Streptococcal toxic shock-like syndrome. Clinical and microbiological aspects. PMID- 9331617 TI - Role of streptococcal proteinase in acute post-streptococcal glomerulonephritis. PMID- 9331618 TI - Arcanobacteriosis. An infection with streptococcus-like features. PMID- 9331619 TI - Non-steroidal anti-inflammatory drugs (NSAIDs). A predisposing factor for streptococcal bacteraemia? PMID- 9331620 TI - The occurrence of nephritis plasmin binding protein (SPEB) in the extracellular products of group C nephritogenic Streptococcus zooepidemicus. PMID- 9331621 TI - Anti-nuclear antibodies in Sydenham's chorea. PMID- 9331622 TI - Microbial associations and response to antimicrobials seen in a psoriasis clinic. PMID- 9331623 TI - Infective endocarditis-inducing abilities of "Streptococcus milleri" group. PMID- 9331624 TI - Isolation and characterization of "Streptococcus milleri" group strains from oral and maxillofacial infections. PMID- 9331625 TI - Serotypes of "Streptococcus milleri" group. PMID- 9331626 TI - "Streptococcus mutans" group in human saliva. Distribution of the different species and susceptibility to antimicrobial agents. PMID- 9331627 TI - Recognition of mutans streptococci by monoclonal antibodies. PMID- 9331628 TI - Transmission of "Streptococcus mutans" in nursing bottle caries and cleft palate patients. PMID- 9331629 TI - Changes in the oral streptococcal flora following irradiation/chemotherapy for bone marrow transplantation. PMID- 9331630 TI - Canine toxic-shock syndrome. An emerging disease? PMID- 9331631 TI - Streptococcal toxic shock syndrome (STSS). An update: a roundtable presentation. PMID- 9331632 TI - Invasive infections caused by Streptococcus pyogenes in Denmark 1990-1994. PMID- 9331633 TI - Group A streptococcal invasive disease in England and Wales. PMID- 9331634 TI - Group A streptococcal outbreak of perianal infection in a day-care centre. AB - Within 9 days in a day-care centre of 20 children 3 cases and 2 suspected cases of perianal infection (PAI) were revealed. The causative agent for PAI was confirmed to be beta-haemolytic group A streptococcus (GAS). The isolates were further characterized by T- and M-serotyping, conventional restriction endonuclease analysis (REA) and pulsed field gel electrophoresis (PFGE). The typing methods revealed that all outbreak isolates were of same T28R28-serotype and genetically identical. Unrelated, control GAS isolates of the same serotype differed genetically from the outbreak isolates indicating that a single clone had caused the perianal infections. PMID- 9331635 TI - Analysis of active surveillance and passive notification of streptococcal diseases in the Czech Republic. PMID- 9331636 TI - Restriction enzyme analysis (REA) of group A streptococcal (GAS) M-serotypes 1, 3, and 28. A comparison of isolates from severe systemic infections (SSI) and from uncomplicated pharyngitis (UP): epidemiologic and pathogenetic implications. PMID- 9331637 TI - Epidemiological monitoring of group A streptococcal (GAS) infections. PMID- 9331638 TI - Distribution of the serotypes of group A streptococci in Bulgaria from 1968 till 1996. PMID- 9331639 TI - Group A streptococcal infections in France. Clinical features and epidemiological markers. The Groupe d'Enquete 1995 sur les Infections Streptococciques. PMID- 9331640 TI - Clinical epidemiology of rheumatic fever and rheumatic heart disease in tropical Australia. PMID- 9331641 TI - Distribution of group A serotypes and streptococcal pyrogenic exotoxin gene types isolated from blood in Canada. PMID- 9331642 TI - Serotype, biotype, pyrogenic exotoxin, streptolysin O and exoenzyme patterns of invasive Streptococcus pyogenes isolates from patients with toxin shock syndrome, bacteremia and other severe infections. PMID- 9331643 TI - Cases of streptococcal toxic shock syndrome in Germany since 1989, toxin (mitogen) expression, content of toxin genes and relation to M-serotypes. PMID- 9331644 TI - Vaginal colonization by Streptococcus agalactiae in two neighbouring populations in Tuscany. PMID- 9331645 TI - Group B streptococcal infections in neonates and its carriage in women. PMID- 9331646 TI - The epidemiology of beta-haemolytic streptococcal infections in the Intensive Therapy Unit of the Royal Infirmary, Edinburgh 1991-1994. PMID- 9331647 TI - Epidemiology of group B streptococcal infection in a special care neonatal unit. PMID- 9331649 TI - Combined, selective chemoprophylaxis of early onset neonatal group B streptococcal disease (GBS EOD). PMID- 9331648 TI - Obstetric management and group B streptococcal sepsis. Have we made progress? Has it made a difference? PMID- 9331650 TI - Serotyping distribution and antimicrobial resistance of Streptococcus pneumoniae isolated in Brazil (1992-1996). PMID- 9331651 TI - Serotype distribution of clinical isolates of Streptococcus pneumoniae from a Delhi hospital. PMID- 9331653 TI - Characterization of enterococci isolated from nosocomial and community infections in Brazil. PMID- 9331652 TI - Dynamics of Enterococcus faecalis colonization of bone marrow transplant patients. PMID- 9331654 TI - Biotype distribution of enterococci and group D streptococci recovered from clinical material. PMID- 9331656 TI - Epidemiology of vancomycin-resistant Enterococcus faecium infections in a liver transplant unit. PMID- 9331655 TI - Isolation of vancomycin-resistant enterococci from supermarket poultry. PMID- 9331657 TI - Taxonomy and epidemiology. Molecular approaches: an opening lecture. PMID- 9331658 TI - Application of emm gene sequencing and T antigen serology for typing group A streptococcal systemic isolates. Survey of random and outbreak-related isolates. PMID- 9331659 TI - High-resolution genotyping of Streptococcus pyogenes. Application to outbreak studies and population genetics. PMID- 9331660 TI - Molecular epidemiology of group A streptococcal infection in the Northern Territory of Australia. PMID- 9331661 TI - A novel method for the primary diagnosis of group A streptococci from clinical specimens. PMID- 9331662 TI - A novel method for the serodiagnosis of group A streptococcal antibodies. PMID- 9331664 TI - Anomalies in emm typing of group A streptococci. PMID- 9331663 TI - Clonal analysis of five M types causing most disease in New Zealand. PMID- 9331665 TI - Improvement of biological support for the community control of rheumatic fever in Algeria. PMID- 9331666 TI - DNA restriction profiles of nontypable group B streptococcal clinical isolates. PMID- 9331667 TI - Rapid and specific diagnosis of group B streptococcal infection by the polymerase chain reaction (PCR). PMID- 9331668 TI - Analysis of pathogenic group B streptococci by pulsed field gel electrophoresis. PMID- 9331669 TI - An apparently new strain-variable Streptococcus agalactiae protein. PMID- 9331670 TI - PCR for detection of Streptococcus equi. PMID- 9331671 TI - Human isolates of large colony-forming beta hemolytic group G streptococci form a distinct clade upon 16S rRNA gene analysis. PMID- 9331673 TI - Chromosomal markers for the molecular typing of antibiotic-resistant Streptococcus pneumoniae strains. PMID- 9331672 TI - Intra-specific diversity within Streptococcus anginosus. PMID- 9331674 TI - Molecular typing of Streptococcus pneumoniae strains isolated in Romania. PMID- 9331675 TI - Reported single clone multiple antimicrobial-resistant 23F Streptococcus pneumoniae can be differentiated by pulsed-field gel electrophoresis (PFGE). PMID- 9331676 TI - Use of a DNA probe test for identification of Streptococcus pneumoniae nontypable strains. PMID- 9331677 TI - PCR typing of Enterococcus faecium. An evaluation. PMID- 9331678 TI - Streptococcus infantarius sp. nov. related to Streptococcus bovis and Streptococcus equinus. PMID- 9331679 TI - Recent approaches on the taxonomy of the enterococci and some related microorganisms. PMID- 9331680 TI - Molecular analysis of Lactococcus garvieae and Enterococcus gallinarum isolated from water buffalos with subclinical mastitis. PMID- 9331681 TI - Comparison of PCR with phenotypic methods for the speciation of enterococci. PMID- 9331682 TI - Investigation by long PCR of the genetic elements mediating VaNa glycopeptide resistance in enterococci from uncooked meat in south Manchester. PMID- 9331683 TI - Typing methods for a study of gene transfer in enterococci. PMID- 9331684 TI - Identification of enterococcal strains using an inductive learning algorithm. PMID- 9331685 TI - Antibiotic resistance in streptococci and enterococci. Where are we, where are we going? An opening lecture. PMID- 9331686 TI - Genetic studies of cefotaxime resistance in Streptococcus pneumoniae. PMID- 9331687 TI - Clindamycin in recurrent group A streptococcal pharyngotonsillitis. An alternative to tonsillectomy? PMID- 9331688 TI - Lack of penicillin tolerance in group A streptococci. PMID- 9331689 TI - Erythromycin resistance phenotypes in Swedish clinical isolates of group A streptococci. PMID- 9331690 TI - Group A streptococci. Erythromycin resistance and penicillin tolerance. PMID- 9331692 TI - The study of penicillin tolerance in Streptococcus pyogenes. PMID- 9331691 TI - Guidelines for detection of penicillin tolerance in Streptococcus pyogenes by MIC MBC method. AB - Disagreement on a uniform and accepted method of testing penicillin tolerance in Streptococcus pyogenes prevents a meaningful comparison of results of various studies on tolerance and an assessment of the contribution of this phenomenon to treatment failure. Therefore guidelines are needed. PMID- 9331693 TI - Penicillin tolerance in group A streptococci. PMID- 9331694 TI - Differential response to benzylpenicillin-G in tolerant and nontolerant strains of Streptococcus pyogenes in human PMNL and epithelial cells in vitro. PMID- 9331695 TI - Susceptibility of beta-haemolytic streptococci to penicillin. PMID- 9331696 TI - Study of resistant pneumococci in Romania between 1973-1995. PMID- 9331697 TI - Association of transposon Tn1545 with multidrug resistant strains of Streptococcus pneumoniae isolated in Canada. PMID- 9331698 TI - Genetic and molecular characterization of high-level gentamicin resistance in Enterococcus faecalis strains isolated in Romania. PMID- 9331699 TI - The in-vitro activity of LY333328, a new glycopeptide, against clinical isolates of vancomycin-resistant enterococci. PMID- 9331700 TI - Study of Enterococcus faecium BM6226. A strain resistant to most antibiotics used in enterococcal antibiotherapy. PMID- 9331701 TI - Resistance to lincosamides by nucleotidylation associated with conjugative transfer of a large chromosomal element in Enterococcus faecium. PMID- 9331702 TI - Analysis of regulatory region of ermAM genes in streptococci and enterococci highly resistant to macrolides and lincosamides. PMID- 9331703 TI - Cell-to-cell signalling between group A streptococci and pharyngeal cells. Role of streptococcal surface dehydrogenase (SDH). PMID- 9331704 TI - Human C4BP binds to the hypervariable N-terminal region of many members in the streptococcal M protein family. PMID- 9331705 TI - Proteins M6 and F1 are required for efficient invasion of group A streptococci into cultured epithelial cells. AB - Group A streptococci were recently shown to be capable of invading human epithelial cell monolayers. Cell invasion might be an important virulence trait of streptococci that enable the pathogen to gain entry into deeper tissues after initial binding to host cells. Nothing is known concerning the nature of streptococcal components that mediate invasion. Using isogenic mutants of strain JRS4 that are defective in the expression of either M6 protein or protein F1, or both proteins, it was demonstrated that both adhesins are required for efficient invasion. Further more, expression of protein F1 on the surface of a non-invasive strain rendered the latter invasive, suggesting that protein F1 is directly involved in the invasion process. PMID- 9331706 TI - Hyaluronic acid capsule modulates interactions of group A streptococci with human epidermal keratinocytes. PMID- 9331707 TI - Novel superantigens from streptococcal toxic shock syndrome Streptococcus pyogenes isolates. PMID- 9331708 TI - Circulating anti-IgG and glomerular damage in rabbits immunized with IgG Fc receptor positive group A streptococci. PMID- 9331710 TI - The M6 protein of Streptococcus pyogenes and its potential as a tool to anchor biologically active molecules at the surface of lactic acid bacteria. PMID- 9331709 TI - Molecular markers for throat and skin isolates of group A streptococci. AB - In summary, the emm chromosomal patterns distinguish between the two principal tissue site reservoirs of group A streptococci--the nasopharyngeal mucosa and impetigo lesion. Strains derived from normally sterile tissue sites are probably transmitted to new hosts by respiratory droplets, at least in the Connecticut population. The speA gene provides an example of how genetic exchange between different strains of group A streptococci may be limited to a single tissue site or to a subset of emm chromosomal patterns. PMID- 9331711 TI - Effects of environmental factors on streptococcal erythrogenic toxin A (SPE A) production by Streptococcus pyogenes. PMID- 9331712 TI - Superantigens of group A streptococci (Streptococcus pyogenes). Distribution, induction of antibodies, and binding to human cell types. PMID- 9331713 TI - Group A streptococcal M protein binds to several human cell types but not via MHC class II molecules. PMID- 9331714 TI - Superantigenic activity produced by group A streptococcal isolates is neutralized by plasma from IVIG-treated streptococcal toxic shock syndrome patients. PMID- 9331715 TI - Physicochemical separation of A beta subtype from A alpha type erythrogenic toxin produced by T1 strain Streptococcus pyogenes. PMID- 9331716 TI - A new exocellular mitogen from NY5 strain Streptococcus pyogenes antigenically different from erythrogenic toxins A, B, and C. PMID- 9331717 TI - Mapping of binding sites for human serum albumin and fibrinogen on the M3 protein. Molecular model and function in the pathogenic mechanism. PMID- 9331718 TI - Binding of C1q to group A streptococcal M-family proteins. PMID- 9331719 TI - Antigenic diversity of IgG Fc-receptors in Streptococcus pyogenes. PMID- 9331720 TI - Structure-function and pathogenesis studies of Streptococcus pyogenes extracellular cysteine protease. AB - Replacement of the single cysteine residue (C192) with serine in the Streptococcus pyogenes extracellular cysteine protease (SCP) prevented auto catalytic processing of the 40-kDa zymogen to the 28-kDa mature form and eliminated proteolytic activity. SCP incubated with human endothelial cells induced a time- and concentration-dependent increase in a 66-kDa gelatinase/type IV collagenase in culture supernatants. Activation of this gelatinase/collagenase may contribute to endothelial cell damage, tissue destruction, and hemodynamic derangement observed in some patients with severe, invasive S. pyogenes infection. PMID- 9331721 TI - Streptococcal recombinant receptors as a diagnostic tool. PMID- 9331722 TI - A novel plasminogen/plasmin binding protein on the surface of group A streptococci. PMID- 9331723 TI - Studies on streptococcal NAD-glycohydrolase. Copurification of streptodornase A. PMID- 9331724 TI - Host cell specific adhesins of group A streptococci. PMID- 9331725 TI - Studies on variation in Lancefield group A type 50 streptococcal cell-wall carbohydrate. PMID- 9331726 TI - Clonal invasion of the equine respiratory tract by Streptococcus zooepidemicus. PMID- 9331727 TI - Cloning and expression in Escherichia coli of a protective surface protein from type V group B streptococci. AB - This report describes a trypsin-resistant laddering protein purified from a type V strain, a serotype of important emerging clinical significance. This protein is present in a majority of type V clinical strains, elicits protective antibody in an animal model, and is cross-reactive with the alpha C protein and R1. The gene encoding this protein has been cloned; preliminary nucleotide sequence analysis reveals significant homology, though not identity, with the alpha C protein gene. These data support the hypothesis that there exists a family of related but distinct GBS surface proteins which may play a role in immunity to GBS infection. PMID- 9331728 TI - The Rib and alpha proteins define a family of group B streptococcal surface proteins that confer protective immunity. PMID- 9331729 TI - Glycolipid intermediates in biosynthesis of group B streptococcal capsular polysaccharide. PMID- 9331730 TI - The role of group B streptococci beta-hemolysin expression in newborn lung injury. AB - There is a direct correlation between the level of GBS beta-hemolysin expression and the ability of GBS to injury lung epithelial cells. Electron microscopy suggest the hemolysin acts as a pore-forming cytolysin. beta-hemolysin-associated lung epithelial cell injury is inhibited by surfactant phospholipid, a substance in which high-risk premature infants are deficient. We have now shown that loss of GBS hemolysin activity is associated with decreased animal virulence following intrathoracic inoculation of the organism. Further, a knockout of a putative GBS beta-hemolysin gene from the literature suggests it is not the major GBS hemolysin determinant. Cloning and sequencing analysis of the Tn916 (or Tn916DE) insertions in three of our nonhemolytic GBS mutants show identical integration sites in a distinct chromosomal locus. Finally, a putative 11-kd hemolysin species is identified by comparative analysis of protein extracts from isogenic hemolysin mutants. PMID- 9331731 TI - Epithelial cell invasion by group B streptococci is important to virulence. PMID- 9331732 TI - Surface protein expression in group B streptococcal invasive isolates. AB - Results from characterization of 211 GBS isolates from early-onset disease indicated that serotypes Ia, III and V accounted for almost 80% of the isolates, and that alpha was the protein most often expressed. Each of the common polysaccharide types had a characteristic predominant protein expression pattern: alpha for Ia, R4 for type III and R1+R4 for type V isolates. Expression of alpha protein was always mutually exclusive of R proteins. The presence of more than one species of R by a given isolate was confirmed by IEP. In addition, PAGE/WB studies verified the multiple MW forms of R1, and the variation from strain to strain in the highest form of R4 that we had previously reported. Our data not only showed the great complexity of the GBS cell surface but also demonstrated the advantage of using both type polysaccharides and surface-localized proteins as markers for characterization of GBS strains. PMID- 9331733 TI - Streptococcus pneumoniae choline binding proteins. Role in cell wall turnover. PMID- 9331734 TI - Characterization of a novel plasminogen activator from Streptococcus uberis. PMID- 9331735 TI - The auxotrophic nature of Streptococcus uberis. The acquisition of essential acids from plasmin derived casein peptides. PMID- 9331736 TI - Virulence markers of Streptococcus suis type 1 and 2. PMID- 9331737 TI - Determination of the transcript size and start site of the putative sca operon of Streptococcus gordonii ATCC 51656 (formerly strain PK488). PMID- 9331739 TI - Modification of sucrose dependent cell adherence by deletion and reintroduction of the gtf genes in Streptococcus mutans. PMID- 9331738 TI - Regulation of Streptococcus gordonii glucosyltransferase. PMID- 9331740 TI - Induction of NO production by polyosides from the wall of Streptococcus mutans OMZ175 in the rat aorta. PMID- 9331741 TI - Genetic analysis of the surface protein antigen gene expression in Streptococcus mutans. PMID- 9331742 TI - Chromosomal deletions in Streptococcus mutans. PMID- 9331743 TI - The role of hyaluronidase in growth of Streptococcus intermedius on hyaluronate. PMID- 9331744 TI - Possible pathogenic effect of Streptococcus mitis superantigen on oral epithelial cells. PMID- 9331745 TI - Purification and partial characterization of a novel human platelet aggregation factor in the extracellular products of Streptococcus mitis, strain Nm-65. PMID- 9331746 TI - Identification and further characterization of a locus coding for a hypothetical 33.6-kDa protein involved in intrageneric coaggregation of oral streptococci. PMID- 9331747 TI - The N-terminal half part of the oral streptococcal antigen I/IIf contains two distinct functional domains. PMID- 9331748 TI - Function and immunogenicity of cell-wall-anchored polypeptide CshA in oral streptococci. PMID- 9331749 TI - The glucan binding domain of the Streptococcus mutans glucan binding protein. PMID- 9331750 TI - Cell-adherent glucan does not protect endocarditis-causing viridans streptococci against bactericidal proteins from human blood platelets. PMID- 9331751 TI - Sialic acid utilisation by viridans streptococci. PMID- 9331752 TI - Protein I/II from oral viridans streptococci modulates expression of E-selectin, ICAM-1 and VCAM-1, and promotes transendothelial migration of neutrophils in vitro. PMID- 9331753 TI - Binding of enterococci to extracellular matrix proteins. PMID- 9331754 TI - M protein expression is not required for resistance to phagocytosis of type 18 group A streptococci. PMID- 9331755 TI - Detection of the erythrogenic toxin A, B, and C genes in group A streptococci isolated from clinical specimens. PMID- 9331756 TI - Cloning of a chromosomal region responsible for streptolysin S production in Streptococcus pyogenes. PMID- 9331757 TI - Identification and characterization of a new protein from Streptococcus pyogenes having homology with fibronectin and fibrinogen binding proteins. AB - Fibronectin and fibrinogen-binding proteins have been described as possible adhesin in streptococci and staphylococci. Recent published data has demonstrated that Protein F, a fibronectin-binding protein from group A streptococci, is important in adherence to respiratory cells (8). Other similar proteins already described (i.e. SOF, Sfb and SfbII) are able to competitively inhibit the binding of fibronectin to S. pyogenes (9,10,5). Similarly, clumping factor from S. aureus, is known to promote adherence to fibrinogen-coated surfaces (7). When the sequence from SFFBP-12 was compared against all the others fibronectin and fibrinogen-binding proteins described in streptococci and staphylococci (1-10), an identity at the amino acid level, ranging from 38 to 69% was found for the C region. For the repeated region (R), the identity ranged between 47 and 75%. Unlike all the other proteins already described in group A streptococci, the protein we describe here, SFFBP-12, shares a high degree homology (67-75%) with the fibronectin-binding protein B from S. dysgalactiae (6), as well as homology with the S. aureus clumping factor (7) and fibronectin-binding protein B (3), making it a new potential fibronectin-fibrinogen binding protein for group A streptococci. These characteristics would also imply that SFFBP-12 contains two different fibronectin-binding domains (regions B and C), thus enhancing its role as a possible major adhesin molecule. RNA transcription assays showed a transcript with the expected molecular size for the intact SFFBP-12 protein, confirming that the protein is actively expressed during bacterial growth. SFFBP 12 is the largest protein of its kind identified from group A streptococci and is comparable in size to the fibronectin binding protein B from S. dysgalactiae (6). If it is shown that SFFBP-12 does in fact bind both fibronectin and fibrinogen, as the sequence data suggest, it would make this molecule a major virulence determinant for the group A streptococcus. PMID- 9331758 TI - Dual affinity of streptococcal protein F for fibronectin and fibrinogen. PMID- 9331759 TI - M proteins of Streptococcus equisimilis strains isolated from pharyngitis patients. PMID- 9331760 TI - Genetic mosaic upstream of scpG in human group G streptococci contains sequences from group A streptococcal virulence regulon. PMID- 9331761 TI - Albumin receptor protein of group G Streptococcus. Cloning of the gene, characterization, and usage of the protein expressed in Escherichia coli. PMID- 9331762 TI - Structural and functional similarity of C5a-ase enzymes from group A and B streptococci. PMID- 9331763 TI - Sequence heterogeneity in the capsule production locus of Streptococcus pneumoniae. PMID- 9331764 TI - Selection system for the isolation of in vivo activated promoters from endocarditis-causing viridans streptococci. PMID- 9331765 TI - A conserved region of a hyaluronidase gene from Streptococcus intermedius. PMID- 9331766 TI - Intermedilysin. A cytolytic toxin specific for human cells of a Streptococcus intermedius isolated from human liver abscess. PMID- 9331767 TI - Identification of virulence genes in Enterococcus faecalis by differential display polymerase chain reaction. PMID- 9331768 TI - In vivo survival of Enterococcus faecalis is enhanced by extracellular superoxide production. PMID- 9331769 TI - Regulation of aggregation substance expression by bacterial and host factors. PMID- 9331770 TI - Aggregation and binding substances enhance pathogenicity in a rabbit model of Enterococcus faecalis endocarditis. PMID- 9331771 TI - Virulence of Streptococcus pyogenes for chicken embryos after isogenic inactivation of different streptococcal pathogenicity factors. PMID- 9331772 TI - A critical role of tumor necrosis factor (TNF) alpha in experimental group A streptococcal (GAS) bacteremia. PMID- 9331773 TI - Expression of activational markers on circulating leukocytes from baboons with group A streptococcal (GAS) bacteremia. PMID- 9331774 TI - Erythrogenic toxin-induced arteritis in a rabbit ear model. Comparison with Arthus reaction angiitis. AB - We have developed a local type experimental model of angiitisin rabbits. Repeated intracutaneous injections of erythrogenic toxins types A and C (ETA, ETC) along the intermediate auricular artery of the rabbit ear produced subacute type arteritis, characteristic of lymphocyticinfiltration, simulating Kawasaki disease angiitis. Staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin 1 (TSST) also induced similar lesions, indicating superantigens as inflammogens in vivo. Conversely, the Arthus reaction included acute type angiitis when tested similarly in rabbits immunized to human serum albumin. The ear artery was infiltrated by heterophil leukocytes, often together with venules and capillaries affected, resembling periarteritis nodosa and leukoclastic vasculitis in human disease. PMID- 9331775 TI - An experimental model for acute post-streptococcal glomerulonephritis in mice. PMID- 9331776 TI - An experimental model of group G streptococcal soft-tissue infections. Studies of acquired immunity. PMID- 9331777 TI - Group B streptococci. Role of capsular polysaccharide on virulence and induction of septic arthritis. PMID- 9331779 TI - Virulence mechanisms of Streptococcus suis. PMID- 9331778 TI - Virulence of strains of new types and type candidates of group B streptococci (Streptococcus agalactiae). Comparative evaluation using mice and a chicken embryo model. PMID- 9331780 TI - Murine and pig models of Streptococcus suis type 2 infections are incompatible. PMID- 9331781 TI - Assessment of bacterial physiology in the digestive tract by use of luciferase gene as promoter probe. PMID- 9331782 TI - Protection against infections with Streptococcus pneumoniae and Haemophilus influenzae type B in a population of splenectomized individuals with varying vaccination status. PMID- 9331783 TI - Immune response to the 23-valent pneumococcal polysaccharide vaccine in lymphoma patients and patients with chronic renal diseases. PMID- 9331784 TI - Bactericidal activity elicited by the beta C protein of group B streptococci contrasted with capsular polysaccharides. PMID- 9331785 TI - Combination conjugate vaccines against multiple serotypes of group B streptococci. PMID- 9331786 TI - Vaccination with highly purified cell surface proteins confers protection against experimental group B streptococcal infection. PMID- 9331787 TI - Immunization with a single-repeat alpha C protein may prevent escape of lower repeat mutants of group B Streptococcus. PMID- 9331788 TI - Intranasal immunisation of mice with a streptococcal peptide-based vaccine. PMID- 9331789 TI - Group A and group B streptococcal vaccine development. A round table presentation. AB - The data presented above provide a broad overview of ongoing work to develop vaccines against group A and group B streptococcal infections. The encouraging results of human trials with conjugate group B polysaccharide vaccines suggest that this approach will lead to a safe and effective method for preventing these devastating infections in newborn infants. The results of preclinical studies of the various strategies to develop group A streptococcal vaccines are also encouraging. Whether one approach will be more advantageous or efficacious than another will need to await clinical trials. Nevertheless, we predict that in the next decade we will make significant strides in preventing streptococcal infections and their complications. PMID- 9331790 TI - Cytokines in streptococcal infections. An opening lecture. PMID- 9331791 TI - Salivary natural antibodies as a basic immune barrier against group A streptococci. PMID- 9331792 TI - Immunological crossreactivity between the class I epitope of streptococcal M protein and myosin. PMID- 9331793 TI - Cytokine production in an ex vivo whole blood model following induction by group B streptococcal polysaccharides and lipoteichoic acid. PMID- 9331794 TI - Activation of granulocytes by phorbol-12-myristate-14-acetate (PMA) enhances phagocytosis of Streptococcus pyogenes. PMID- 9331795 TI - Neutralization of streptococcal pyrogenic exotoxins and staphylococcal enterotoxins by antiserum to synthetic peptides representing conserved amino acid motifs. PMID- 9331796 TI - Immunoglobulins inhibit adherence and internalization of Streptococcus pyogenes to human pharyngeal cells. AB - Purified human sIgA against group A streptococci inhibited streptococcal adherence to pharyngeal cells whereas rabbit serum raised against the whole M+ strain did not. Of note, inhibition of adherence by sIgA occurred despite a much lower immunreactivity against the M6 protein as compared to the hyperimmune serum against the whole M+ strain. The M protein does probably not mediate the adherence of group A streptococci to respiratory cells, as shown by previous studies. However, since the isogenic M- derivative was less invasive to human pharyngeal cells than the M+ strain, our invasion experiments suggest that the M protein may play a role in internalization of group A streptococci into eukaryotic cells. More importantly, internalization and not adherence could be blocked by rabbit serum immunized with recombinant M6 protein. PMID- 9331797 TI - Human secretory immune response to streptococcal M protein. Identification of a secretory IgA epitope in the exposed conserved region. PMID- 9331798 TI - Phagocytic, serological, and protective properties of streptococcal group A carbohydrate antibodies. AB - Group A streptococcal antibodies promote phagocytosis of several different strains of GR A streptococci. These antibodies passively protect against an in vivo mouse challenge model. PMID- 9331799 TI - Invasive group A streptococcal disease. Association with a lack of anti-exotoxin antibodies. PMID- 9331800 TI - Nonneutralizing antibody to erythrogenic toxin A alpha(NY5ETA) in human sera. AB - A solid phase adsorption experiment was performed to detect anti-ETA alpha(NY5 strain ETA) nonneutralizing antibody. Toxin was applied to the Protein A Sepharose column retaining IgG bound after pretreatment with test serum. Mitogenic activity recovered in the effluent and in the eluate containing the IgG was measured separately in rabbit lymphocyte culture. A significantly increased recovery in the eluate was found in combination with decreased recovery in the effluent of three sera from Kawasaki disease and one from Streptococcus pyogenes infection among 13 ELISA-positive, antimitogen assay (AMA)-negative sera tested. The nonneutralizing antibody may play an important role in the immune protection against ETA by binding to nonmitogenic epitope(s) on ETA and enabling to handle the toxin not as a superantigen but as a conventional peptide antigen. PMID- 9331801 TI - Streptolysin O modulates cytokine synthesis in human peripheral blood mononuclear cells. PMID- 9331802 TI - Cytokine profile of human peripheral blood mononucleated cells stimulated with a novel streptococcal superantigen, SPEA, SPEC and group A streptococcal cells. PMID- 9331803 TI - Cytokine networks in Sydenham's chorea and PANDAS. PMID- 9331804 TI - ELISA for detecting proteinases produced by group A streptococci. PMID- 9331805 TI - Role of complement and complement receptor C1qR in the antibody-independent killing of group B streptococcus. PMID- 9331806 TI - Age-related sensitivity of neonatal mice to toxicity induced by heat-killed group B streptococci. PMID- 9331807 TI - Influenza A-group B streptococcal mixed infection. Study of systemic humoral immunity. PMID- 9331808 TI - Effect of C5a and tumor necrosis factor-alpha on phagocytosis of Streptococcus agalactiae NT/X and IV/X by bovine neutrophils. PMID- 9331810 TI - The Streptococcus pyogenes genome sequencing project. A progress report. PMID- 9331809 TI - Changes in bovine neutrophils induced by the capsule of Streptococcus uberis. PMID- 9331811 TI - Analysis of hyaluronic acid capsule expression in group A streptococci. PMID- 9331812 TI - Genetic studies of erythrogenic toxin carrying temperate bacteriophages of Streptococcus pyogenes. PMID- 9331813 TI - Regulation of hyaluronic acid capsule production by the has operon promoter in group A streptococci. PMID- 9331814 TI - Chromosomal analysis of group A streptococci by pulsed field gel electrophoresis. PMID- 9331815 TI - Evidence for a site specific genomic rearrangement in the slo region of Streptococcus pyogenes. PMID- 9331817 TI - Elicitation of high (p)ppGpp levels in Streptococcus equisimilis without imposing nutritional deprivation. PMID- 9331816 TI - Biological significance of the genetic linkage between streptolysin S expression and riboflavin biosynthesis in Streptococcus pyogenes. PMID- 9331818 TI - Analysis of the capsule synthesis locus, a virulence factor in group B streptococci. PMID- 9331819 TI - Aberrations in expression of the beta antigen of Streptococcus agalactiae. PMID- 9331820 TI - Development of a group B Streptococcus (GBS) cloning system. PMID- 9331821 TI - Characterization of the stress response in Streptococcus pneumoniae. PMID- 9331822 TI - Molecular characterization of a gene locus encoding biosynthesis pathway of Streptococcus mutans serotype C-specific antigen. PMID- 9331823 TI - Regulation of the galactose operon of Streptococcus mutans. PMID- 9331824 TI - Competence-pheromone in Streptococcus sanguis. Identification of the competence gene comC and the competence pheromone. PMID- 9331825 TI - Genetic analysis of Enterococcus faecalis chromosome carrying mobile elements. PMID- 9331826 TI - TN5252: a model for complex streptococcal conjugative transposons. PMID- 9331827 TI - Analysis of pheromone binding and pheromone reception by Enterococcus faecalis. PMID- 9331828 TI - The pAD1 sex pheromone response in Enterococcus faecalis. PMID- 9331829 TI - Enterococcal pheromone-like activity derived from a lipoprotein signal peptide encoded by a Staphylococcus aureus plasmid. PMID- 9331830 TI - Characterization of the Enterococcus faecalis alpha C protein homolog. Evidence for the expression of alternate forms in commensal and infection derived isolates. PMID- 9331831 TI - Identification of a highly conserved lipopolysaccharide (LPS) modification operon in Enterococcus faecalis. PMID- 9331832 TI - Summary of the round table discussion on genome structure and regulation of gene expression in streptococci and enterococci. PMID- 9331833 TI - Endoscopic microsurgical dissection of the esophagus (EMDE). AB - This paper presents endoscopic microsurgical dissection of the esophagus (EMDE), a surgical technique for the therapy of esophageal cancer which improves blunt esophageal dissection with the aim of reducing postoperative morbidity and mortality. A mediastinoscope with integrated operative instrument channel, fibre bundles, optic and rinsing channel has been developed whereby precise and atraumatic esophageal dissection is possible via a cervical access incision. Between 1989 and 1993, 37 patients were operated on using the EMDE technique and are compared with 48 patients operated on during the same period by the thoraco abdominal route. The operative duration was reduced by the new technique, and although the number of severe complications was not significantly different between both groups, the rate of pulmonary and cardiac complications was reduced. The mortality rate was 10% for EMDE patients and 14% for the thoraco-abdominal procedure, and there was no difference in the long-term survival rate. As distinct from procedures requiring a thoracotomy for esophageal dissection, EMDE permits ventilation of both lungs throughout the entire operation and reduces the total operative trauma. PMID- 9331835 TI - Thoracoscopic esophageal surgery. AB - Thoracoscopic surgery has been utilized for both esophageal benign lesions and malignant lesions. Resection of submucosal and mural benign lesions, as well as Heller Myotomy, are prime indications for the procedure. Resection of or staging procedures have been performed by various centers around the world for esophageal carcinoma. Resection of epiphrenic diverticula and performance of antireflux endoscopic surgical procedures are indications for the VATS procedure. Transthoracic thoracoscopic vagectomy has been performed for a number of years. Avoidance of perforation of the esophageal mucosa and subsequent leakage are prime concerns of thoracoscopic video assisted surgery on the esophagus. PMID- 9331834 TI - Surgical treatment of spastic conditions of the esophagus. AB - Primary esophageal motility disorders include achalasia, diffuse and segmental esophageal spasm, nutcracker esophagus and hypertensive lower esophageal sphincter. Failed medical therapy frequently precedes the presentation of these patients for surgical intervention. Both laparoscopic and thoracoscopic techniques have been developed to successfully treat these spastic disorders of the esophagus. Laparoscopic and thoracoscopic operative techniques are described. PMID- 9331837 TI - Does VATS favor seeding of carcinoma of the lung more than a conventional operation? AB - Seeding of carcinoma has always been of concern to surgeons. Recent reports have focused on possible implantation of tumor in the small wounds of minimally invasive procedures, i.e. VATS. A patient is presented who had a conventional open lobectomy for carcinoma and later developed tumor in the traditional thoracotomy wound. Although seeding or tumor implantation is accepted by thoracic surgeons, the exact mechanism of tumor implantation has never been scientifically explained or documented. Possibly, a rare and necessary pattern of genetic mutations could be responsible for this infrequent but serious problem. PMID- 9331836 TI - Present day concepts of thoracoscopy as a modality in pediatric cancer management. AB - Recent improvements in video imaging and instrumentation have encouraged a wider use of thoracoscopy as a modality for diagnostic procedures. Its utility for resection is still being reviewed. To assess the utility, diagnostic accuracy, and morbidity of thoracoscopy in children with cancer, we reviewed the experience at our institution. Between January 1991 and July 1995 sixty-four (64) procedures were performed either to diagnose pulmonary nodules of indeterminate origin (n = 42) or mediastinal masses (n = 11) or to evaluate pulmonary infiltrates in leukemia (n = 11). Thoracoscopy yielded a successful diagnosis in 90% of the cases. Conversion to open thoracotomy was necessary in 11 patients. Thoracoscopy in the management of children with cancer is useful for staging, obtaining diagnostic tissue, and is associated with a low morbidity. PMID- 9331838 TI - Current applications of nonvideo thoracoscopy. AB - Video thoracoscopy has markedly improved the surgical team's ability to visualize the pleural cavity. As a result, traditional nonvideo thoracoscopy techniques have fallen into disuse. Although video technology has caused a resurgence of interest in thoracoscopy, and has made previously unimaginable endoscopic procedures feasible, it is not obviously superior to nonvideo thoracoscopy in all situations. The defining features of nonvideo thoracoscopy include an open tube type of thoracoscope (mediastinoscope), direct visualization of the pleural space through the thoracoscope, and passage of instruments through the thoracoscope. Potential clinical advantages of nonvideo thoracoscopy are related to the following features: simplicity of equipment, low cost, feasibility without one lung ventilation, feasibility under local or regional anesthesia, instrumentation through the thoracoscope (single incision), and ease of irrigation and suction. Nonvideo thoracoscopy is a simple and effective endoscopic technique for diagnostic pleural biopsy, pleurodesis of malignant effusions, debridement and drainage of empyemas, and evacuation of clotted hemothoraces. More complex thoracoscopic procedures require video thoracoscopy. Video capabilities have recently been added to several traditional open tube mediastinoscopes. In this way, it may be possible to combine some features of traditional nonvideo, open tube, thoracoscopy with video thoracoscopy. PMID- 9331839 TI - Comparison of bacterial translocation during traumatic shock and hemorrhagic shock in rats. AB - Traumatic shock has been classified as a kind of hypovolemic shock similar to hemorrhagic shock. Since bacterial translocation has been observed in shock, this study investigated the difference in bacterial translocation during traumatic shock and hemorrhagic shock, and considered this effect on lung injury during sepsis. METHODS: Forty-eight male or female Sprague-Dawley rats were divided into 2 groups, hemorrhagic shock and traumatic shock. Bacterial translocation, endotoxin, and blood gas were evaluated. Alterations of the lungs morphologically and functionally were observed. RESULTS: Traumatic shock induced more bacterial translocation and endotoxemia from the gut. Blood gas analysis shows a more severe disorder in traumatic shock than in hemorrhagic shock. Pathological morphologic changes of lungs were more severe in traumatic shock than in hemorrhagic shock. CONCLUSIONS: Traumatic shock cause more bacterial translocation and endotoxemia which subsequently caused serial pathological alterations in lung morphologically and functionally than pure hemorrhagic shock does. These results suggest that this trauma activates more severe mechanism to damage lungs. PMID- 9331840 TI - Left thoracic approach for cancer of cardia: early and late results. AB - BACKGROUND: The surgical treatment of cancer of the cardia is controversial and results are often disappointing. Concern exists not only with regards to the surgical approach but also to the extent of the resection. The authors analyze their experience over a 20-year period adopting almost exclusively a "limited" esophagogastrectomy with a wide regional lymphadenectomy through a left thoracotomy. The aim of the study is to determine if this approach actually plays a role in the treatment of this tumor. METHODS: 148 patients were evaluated for cardial cancer. Of these 22 (14.8%) were not resectable and 6 (4%) received other types of resections for technical reasons. 120 patients are the basis of the present analysis. More than 75% of patients were in stage III or IV. Follow-up was completed in 92.5% of cases; all surviving patients had at least 5 years of follow-up. RESULTS: Four (3.3%) patients died in the postoperative period. In 6 cases (5%) an anastomotic leakage occurred and this caused the death of 2 patients. Nine (7.5%) patients had severe pulmonary complications. Dysphagia was relieved in all non complicated patients. 13 (10.8%) patients had anastomotic recurrence. Overall survival rate after 5 years was 25.62 +/- 6.1%. A significant difference in survival was noted in patients at stages II and III after 5 years (61.3% vs 18.6, p < 0.02). CONCLUSIONS: This operation has proved to be a good option providing satisfying long-term results and a lower incidence of complications if compared with more extended procedures. It can be performed in the majority of patients with carcinoma of the cardia with a low mortality and morbidity and with excellent palliation of dysphagia. In our opinion it remains an optimum treatment for cardial cancer. PMID- 9331841 TI - Vascular reconstruction for intraoperative major vascular injuries. AB - BACKGROUND: The purpose of this study was to find an effective and precise surgical procedure to cope with the major vascular injuries that might be encountered during hepato-biliary and pancreatic surgery. MATERIALS AND METHODS: Among the 220 cases of hepato-biliary and pancreatic surgery from 1991 to 1995 in our department, 8 patients who sustained 13 different instances of vascular injury were reviewed retrospectively. RESULTS: Five injuries occurred in the portal vein (PV), 4 in the hepatic artery (HA), and 2 in the inferior vena cava (IVC). The repair procedures consisted of 2 direct closures, 2 resections of the injured sites followed by the end to end anastomosis, 2 autologous venous grafts and 1 artificial vascular graft implantation, and 2 diversions of the arterial bloodflow of the middle colic artery and the gastroepiploic artery to the injured HA. The reconstructed vessels showed postoperative obstructions in 2 cases, in which artificial vascular graft implantation and direct closure were used for the reconstruction. CONCLUSIONS: Based on the above findings, the use of an artificial graft and a direct closure all contributed to the early postoperative obstruction of the reconstructed vessels. However, a diversion of the arterial bloodflow to the injured hepatic arteries was found to maintain a good postoperative patency in these cases. PMID- 9331842 TI - Abdominal wall lifting with spinal anesthesia vs pneumoperitoneum with general anesthesia for laparoscopic herniorrhaphy. AB - BACKGROUND: Laparoscopic herniorrhaphy has generally been very successful since any postoperative inguinal pain or tension is considerably less troublesome than after other open methods. The conventional laparoscopic approach in the treatment of inguinal hernia involves the use of pneumoperitoneum and general anesthesia. Nevertheless, some complications can be encountered and the procedure is costly. We, therefore, examined the possibility of using a more practical and cost efficient method. MATERIALS AND METHODS: Based on our findings, we propose the use of a Kirschner lifting wire as a means of separating the abdominal wall during laparoscopic herniorrhaphy. Two Kirschner wires are introduced through the subcutaneous tissue, between the umbilicus and inguinal ligament, and parallel to the inguinal ligament. Furthermore, we recommend the use of spinal anesthesia as a means by which the problems associated with general anesthesia and the potential cardiopulmonary complications of carbon dioxide insufflation, are circumvented. Fifteen cases of inguinal hernia have been treated with this new method and compared to the more conventional procedure of pneumoperitoneum under general anesthesia. RESULTS: Visibility of the operative field when used in the inguinal region was not limited at all, and Kirschner wire is considerably less expensive. The postoperative course for the patients who were operated by the new method was uneventful. CONCLUSIONS: Our results indicate that this new method can be useful for the treatment of inguinal hernia. PMID- 9331843 TI - Splenectomy in cancer gastrectomy: recommendation of spleen-preserving for early stages. AB - BACKGROUND: The prognostic significance of splenectomy in cancer gastrectomy was reevaluated, in terms of postoperative survival rate, retrospectively. MATERIALS AND METHODS: The cumulative survival rates between a splenectomized group (n = 272) and a nonsplenectomized group (n = 192) were compared. RESULTS: The five-, ten- and fifteen-year survival rates were significantly higher in the nonsplenectomized group in total. When the cancer was classified, according to intraoperative gross findings of the degree of serosal invasion, penetration into adjacent organs and metastases, the five-year cumulative survival rate for Stage I and II patients was significantly higher in the nonsplenectomized group. No significant difference was found between the two groups of Stage II in any of the one- to fifteen-year survival rates. The one-year survival rate for Stage IV patients in the splenectomized group was higher than that in the nonsplenectomized group. CONCLUSIONS: The splenectomy should be limited to the Stage III or IV patients with some specific findings. PMID- 9331844 TI - A splenic-inferior mesenteric venous anastomosis prevents gastric congestion following pylorus preserving pancreatoduodenectomy with extensive portal vein resection for cancer of the head of the pancreas. AB - BACKGROUND: In order to prevent gastric congestion after both of the splenic and coronary veins were taken as part of extensive portal vein resection in pylorus preserving (PP) pancreatoduodenectomy (PD), we made a splenic-inferior mesenteric venous (SpIMV) anastomosis. MATERIALS AND METHODS: Four patients underwent PP subtotal PD with such extensive portal vein resection under the diagnosis of pancreas head cancer. The portal vein was reconstructed by end-to-end anastomosis, and the coronary vein was ligated. Since the stump of the splenic vein could not be approximated to the portal or superior mesenteric vein, shunting the splenic venous flow to the inferior mesenteric vein was attempted by making a SpIMV anastomosis in 3 patients and by preserving the SpIMV confluence in a patient. Postoperative celiac angiography showed that venous outflow from the stomach, spleen and remnant pancreas collected into the splenic vein and passed through the SpIMV anastomosis or confluence, and finally drained into the portal vein by inferior mesenteric to superior mesenteric collateral. RESULTS: No remarkable congestion of the stomach was observed. CONCLUSIONS: In conclusion making a SpIMV anastomosis or preserving the SKpIMV confluence is beneficial for preventing gastric congestion following PP PD with extensive portal vein resection for cancer of the head of the pancreas. PMID- 9331845 TI - Impact of minimally invasive surgery on adrenalectomy for incidental tumors: comparison with laparotomic technique. AB - BACKGROUND: To compare laparoscopic and laparotomic techniques for the excision of incidental adrenal tumors. MATERIALS AND METHODS: Twenty patients with silent adrenal tumor underwent adrenalectomy or tumor excision through the laparotomic (LPT Group: 12 cases) or laparoscopic (LPS Group: 8 cases) approach. LPT Group and LPS Group were comparatively analyzed in terms of age, gender, Body Mass Index (BMI), concomitant diseases, previous upper abdominal surgery, tumor side and size, type of operation (excision vs adrenalectomy), associated procedures, and pathology. The two groups were then compared for intra/postoperative complications, length of operation, and postoperative ileus, pain, hospitalization. Comparisons were performed also adjusting for variables (BMI, depression, tumor size) unhomogeneously distributed between the two groups. RESULTS: LPT Group and LPS Group were comparable when evaluated for age, gender, BMI, concomitant diseases (except for depression), previous upper abdominal surgery, tumor side, type of operation, associated procedures, and pathology. Tumor size (LPKT > LPS, p = 0.03) and depression (LPT < LPS, p = 0.04) turned out to be differently distribution between the two groups, and were then considered, as well as the BMI (p = 0.08), in the covariance analysis. Postoperative ileus (LPT vs LPS: 3.5 +/- 0.2 days vs 2.3 +/- 0.2 days, p = 0.006), pain (LPT vs LPS: 3.0 +/- 0.3 days vs 1.7 +/- 0.2 days, p = 0.012), and hospitalization (LPT vs LPS: 7.9 +/- 1.0 days vs 3.5 +/- 0.4 days, p = 0.0002) were statistically shorter in the LPS Group; the length of operation, even if shorter in the LPS Group (LPT vs LPS: 2.9 +/- 0.3 hours vs 2.5 +/- 0.3 hours), did not reach statistical significance (p = 0.12). Results were confirmed by covariance analysis. CONCLUSIONS: Laparoscopy seems a safe and effective approach which permits adrenal surgery with lower surgical stress than laparotomic technique. PMID- 9331846 TI - Tolerance and feasibility of perioperative treatment with interferon-alpha 2a in advanced cancers. AB - BACKGROUND: Major surgery impairs the cellular immune response. In order to stimulate the immunological system during the perioperative period, we have studied the clinical and biological tolerance, and the immunological and histological effects of a perioperative treatment using progressive doses of Interferon-alpha 2a, from the third preoperative day (D-3) until the tenth postoperative day (D10). MATERIALS AND METHODS: Twenty-three patients undergoing a major surgical procedure for advanced cancer were included. The clinical and biological parameters evaluated were the body temperature and the blood cell counts. Immunological effects were evaluated by counting the total number of lymphocytes, lymphocyte subsets, natural killer cells (NK), and by analysis of the NK activity, and lymphokine-activated killer cell (LAK) assay. RESULTS: Hyperthermia was the most toxic effect of Interferon-alpha but the overall toxicity was minor, even at the highest dose level. In the early postoperative period there was a significant decrease in total lymphocytes, and in most lymphocyte subset counts when compared with D-3. Overall NK and LAK activities significantly increased from D-3 to D-1 (p < 0.02). A postoperative decrease in NK activity was noted that was not significant when compared to pretherapeutic values, whereas a significant decrease in LAK activity did occur on D4 despite the interferon treatment (p < 0.03). Since we found a dose-dependent effect on some lymphocyte subsets, there was not a clear dose-dependent effect on NK and LAK activities. CONCLUSIONS: Perioperative alpha 2a administration is a safe treatment in advanced cancer patients that may allow a postoperative preservation of NK activity and a destruction of potential circulating metastatic cells. Further studies are ongoing on perioperative immunotherapy in advanced cancer patients. PMID- 9331847 TI - Relationship between the long-term effects of intraperitoneal chemotherapy and the expression of p53 and p21 in patients with gastric carcinoma at stage IIIa and stage IIIb. AB - BACKGROUND: The relationship between the expression of p53 and p21 in cases of advanced gastric cancer (t3) with lymph node metastasis [n1(+) or n2 (+)] and the long-term effects of continuous hyperthermic peritoneal perfusion (CHPP) immediately after resection was evaluated. MATERIALS AND METHODS: Between 1983 and 1989, 74 patients with gastric cancer [t3, n1(+) or n2(+)] underwent curative gastrectomy and survived without postoperative complications. These patients were followed up for at least six years. RESULTS: Thirty-seven patients died from recurrence of cancer. In 34 patients who had tumors designated p53-/p21+ or p53+/p21+, CHPP reduced the percentage of patients who died from recurrence of cancer from 10/20 (50%) to 3/14 (21.4%, P = 0.184). However, in 40 patients who had tumors designated p53-/p21- or p53+/p21-, the percentage of patients who were treated with or without CHPP and died from recurrence of cancer was the same, 6/10 (60%) and 18/30 (60%). In the case of patients who had p21-positive tumors, the 5-year survival rate of 11 patients who were treated with CHPP (72.7%) was higher than that of 17 patients who were not treated with CHPP (41.2%, p = 0.027). However, in the case of patients with p21-negative tumors, the 5-year survival rate of 10 patients who were treated with CHPP (50%) was not very different from that of 28 patients who were not treated with CHPP (42.9%, p = 0.308). CONCLUSIONS: These results indicate that the CHPP might be effective in preventing the recurrence of cancer in patients with p21-positive regardless of the expression of p53. PMID- 9331848 TI - Local intravenous therapy in chronic inflammatory and vascular disorders of the foot. AB - BACKGROUND: Chronic inflammatory and vascular disorders of the foot frequently present 2 problems: slow improvement, resulting in long-term therapy, and episodes of critical complications that may seriously threaten the foot. Particularly in a foot with compromised perfusion, the systemic administration of therapeutic agents may be insufficient. Retrograde venous perfusion, RVP, is a new approach that was recently employed for treatment of diabetic neuropathic pedal ulcers. MATERIALS AND METHODS: In this paper, RVP was utilized in 24 patients with diabetic or non-diabetic foot disorders (infection, ulcers and critical pedal ischemia). Employed agents were infused according to predetermined regimens and included: antibiotics, deproteinized hemodialysate and PGE1. RESULTS: The target lesions were eliminated in 20 patients and improved in the others. CONCLUSIONS: The presented therapy proved to be a useful adjunct to conservative or surgical treatment in the studied disorders. Its main impact was in shortening the duration of therapy and overcoming critical complications threatening the foot. PMID- 9331849 TI - Application of a new water jet apparatus in open hepatobiliary surgery: hepatic resection, cholecystectomy, common bile duct lavage. AB - The presently accepted technologies for hepatobiliary surgery have some limitations and disadvantages. Use of water jet dissection is rapidly spreading and its indications are continuously being widened. In 1988 we developed a new bimodular jet-cutter "Parenchimotom-01". After the experimental application on 110 dogs, the apparatus was introduced into clinical practice. Between 1992-1995 we have successfully performed 11 liver resections, 14 cholecystectomies, and 2 lavages of the common bile duct on 25 patients. Marked decrease in blood loss, operative time, rate of complications, and length of hospitalization were noted. We suggest the water jet dissection to be a sophisticated and maximally effective instrumental technology that offers numerous advantages over the other techniques so far commonly accepted in hepatobiliary surgery and which may improve the treatment and prognosis in patients with different hepatobiliary disease entities. PMID- 9331850 TI - Aortic blisters: diagnosis and evolution. AB - In order to evaluate the incidence, the diagnostic modalities and significance of blisters of the abdominal aortic aneurysm wall, in a retrospective review, 14 patients (2.6%) having these lesions were identified between 1983 and 1995. At preoperative examination, aortography had less accuracy (1 case = 20%) than CT scan (3 cases = 27.2%) or MRI angiography (6 cases = 85.7%) to detect blisters; others were discovered intraoperatively in the remaining four patients. Most blisters were located on the anterior or antero-lateral wall of aneurysms; its area ranged from 0.8 to 2.6 cm2. One patient with a suspected blister diagnosed at aortography, during chest physiotherapy for his COPD, presented sudden abdominal pain: at urgent laparotomy, an acute contained rupture of a large blister, without extraluminal blood loss, was found. All patients underwent aneurysm repair, with no postoperative deaths. Occurrence of rupture in one patient clearly indicates the natural course of aortic blisters. MRI angiography may accurately detect these lesions; surgical treatment is necessary for preventing imminent rupture. PMID- 9331852 TI - Is the rectum a conduit or storage organ? AB - The rectum is claimed to be a conduit; as it receives the stools, the rectoanal inhibitory reflex is evoked and defecation occurs. However, in many healthy subjects, stools could be palpated in the rectum by digital rectal examination (DRE) without the subject feeling the desire to defecate. The purpose of this communication is to study whether the rectum is a conduit or a storage organ. The study comprised 48 healthy volunteers (mean age 38.4 +/- 15.8 SD years; 30 men, 18 women). Number of stools per week was recorded and DRE was carried out followed by air enema radiography. The subjects had a normal stool frequency of 7.8 +/- 1.4 per week. Most of them last defecated a mean of 6.2 +/- 3.4 hours prior to the test. Stools were palpated in the lower rectum by DRE in 31/48 subjects and by radiography in 12/16. DRE correlated with radiologic examination in 9/12 subjects; in 3 of them, DRE revealed an empty rectum while radiography showed stools in the upper rectum. The 17 subjects with an empty rectum had their last defecation 5.2 +/- 3.6 hours before DRE, and the 31 subjects with palpable stools 15.6 +/- 12.9 hours. In conclusion, the rectum might be considered not simply as a conduit for stools but also as a storage organ. This occurs when it receives from the sigmoid colon an amount of stools too small to evoke the defecation reflex, or when this reflex is neglected due to unfavorable circumstances of defecation. PMID- 9331851 TI - Preoperative natural killer cell activity: correlation with distant metastases in curatively research colorectal carcinomas. AB - The authors investigated whether host immunity contributes to the development of asynchronous distant metastases in colorectal carcinomas. The host immunity was examined 8 times, pre- and postoperatively during a one year period in 77 curatively operated cases. A prospective study was performed using obtained personal data. During the mean follow-up period of 920 days, 13 patients developed distant metastases. Among the immunological parameters, the preoperative natural killer (NK) cell activity differed significantly between the metastases positive and negative groups. On univariate analysis, dichotomous NK activity, presence of nodal metastases, and venous invasion correlated with metastases. The hazard ratios on multivariate analysis were 4.53, 3.82, and 4.81, respectively. No correlation was noted between NK activity and the progression stages of colorectal carcinomas. These data suggested that attenuated preoperative NK activity is an important background factor for the development of asynchronous distant metastases following curative resection of colorectal carcinomas. PMID- 9331853 TI - Jejunal interposition for peptic stenosis of the esophagus following esophagomyotomy for achalasia. AB - Retrospective analysis of the results of esophagojejunogastrostomy in 21 patients with peptic stenosis after esophagomyotomy for achalasia is reported. All patients complained of severe dysphagia. The esophagogram showed the presence of a 2 to 3 cm long stenosis in the lower esophagus with a diameter < 10 mm. Endoscopic dilatation was possible in 18 cases and it was pursued until the passage of the endoscope was possible. Manometry confirmed the presence of an aperistaltic esophagus with incompetent LES in all cases examined. GERD was detected by 24 hour pH-metry in 15/21 patients (71.4%). One patient died because of postoperative cardiopulmonary failure. Other minor complications occurred in 6 patients. During an 11 year mean follow-up good results were achieved in 17 patients (85%), fair in 2 (10%) and poor in 1 (5%), in whom redundant jejunal loop was resected after 8 years. Resective surgery in peptic strictures after esophagomyotomy is the treatment that guarantees the best long-term results. Esophagojejunogastroplasty represents a valid technique. Careful selection of patients and an accurate surgical technique are fundamental to reduce mortality and morbidity. PMID- 9331854 TI - Surgery and outcome of alveolar echinococcosis of the liver: historical comparison of mass screening systems in Japan. AB - The prognosis of patients with alveolar echinococcosis of the liver (AEL) is excellent when the lesions were completely resected, suggesting the significance of mass screening systems. We investigated resectability and prognosis of the disease with historical comparison of screening systems. The patients were classified into three groups: Group A (n = 63) detected by the current screening system, enzyme-linked immunosorbent assay and ultrasonography, since December 1984; Group B (n = 39), detected by a serological screening system performed between 1974 and 1984; and Group C (n = 64), non-screened patients accidentally discovered. The lesions at an early stage were most frequently found in group A, and the complete resection was performed at 74.6% in A, 41.0% in B, and 18.8% in C (p < 0.0008). Five-year survival rate was 100% in A, 74.0% in B, and 69.5% in C, with a significant intergroup difference (p < 0.05). The current system facilitates early diagnosis of the disease, contributing to its resectability and to a better outcome. PMID- 9331855 TI - Transabdominal preperitoneal laparoscopic inguinal herniorrhaphy (TPLIH) under regional anaesthesia. AB - BACKGROUND: In an attempt to investigate whether laparoscopy really is a major advance in the treatment of inguinal hernia, the authors performed laparoscopic transabdominal preperitoneal inguinal herniorrhaphy under regional anaesthesia in 15 consecutive patients, 7 of whom with severe medical conditions contraindicating general anaesthesia. METHODS: In the first 5 patients (Group 1) an epidural anaesthesia was performed, whereas in the following 10 patients (Group 2), fentanyl was added to the epidural anaesthesia, and bupivacaine was administered into the subarachnoid space. RESULTS: Results from Group 1 were poorer than those obtained in Group 2. All patients complained of shoulder pain and discomfort which required the intraoperative administration of analgesics in 7 patients and conversion to open repair in one patient. CONCLUSIONS: Although laparoscopy is a feasible and effective procedure in repairing inguinal hernias, it is not indicated in high-risk patients who can be safely, effectively, and less expensively treated with open tension-free repair techniques under local anaesthesia. PMID- 9331856 TI - Laparoscopic cholecystectomy in Brazil: analysis of 33,563 cases. AB - The authors present the results obtained in 33,563 laparoscopic cholecystectomies performed in Brazil from 1990 to 1995. Data were obtained by mailing questionnaires to 220 Services, 118 of which responded. The features included the year when the service started its activities, patient distribution by sex and age, surgical indications, prophylaxis with antibiotics, use of nasogastric and vesical catheters, technique used to produce pneumoperitoneum, intraoperative cholangiography, management of choledocholithiasis, necessity and causes of conversion to open surgery, surgical time, intra and postoperative complications, time of hospitalization, mortality, patient return to normal activities, and laparoscopic cholecystectomy in pregnancy. Analysis of the results demonstrated that the data are equivalent to those obtained in leading world countries and at times even better in terms of lesion of the main bile duct, time of hospitalization, etc. PMID- 9331857 TI - Influence of monensin on Holstein steers fed high-concentrate diets containing soybean meal or urea. AB - We conducted two growth trials to evaluate the effects of monensin on amino acid sparing. When Holstein steers were fed a 90% concentrate diet supplemented with soybean meal (13.5% CP), the DMI, ADG, and efficiencies of feed and nitrogen utilization were greater than with urea (P < .10). Monensin improved ADG with both nitrogen supplements (P < .01), but the positive effects of monensin on efficiencies of feed (P = .12) and nitrogen (P = .26) utilization were greater for soybean meal than for urea. Increasing amounts of monensin (0, 11, or 22 mg/kg of DM) caused a linear increase in DMI with urea. Diets with soybean had greater intakes than diets with urea (P < .01); the greatest intake was of a soybean diet with monensin at 11 mg/kg of DM. Holstein steers fed soybean meal at 13.5% CP had lower DMI and greater efficiencies of feed and nitrogen utilization than steers fed 16.7% CP (P < .10). Crude protein level had no effect on ADG (P > .10). Monensin always increased the efficiencies of feed and nitrogen utilization (P < .05), but these trends were greater for diets with 16.7 than for those with 13.5% CP. Overall, monensin decreased DMI (P < .01), but this effect was greater for 16.7% than for 13.5% CP. Because the positive effects of monensin on diet NEg (P = .16) and efficiency of nitrogen utilization (P = .26) were greater for soybean meal than for urea, it seemed that monensin was sparing amino acids. PMID- 9331858 TI - Influence of various factors on farrowing rate on farms using early weaning. AB - A database containing 12,110 farrowing records from 16 commercial farms was abstracted from the PigCHAMP production information system. The farms were selected based on the willingness of producers to keep records and average weaning age. The ADFI records of 10,846 of these lactating sows were also obtained. Sows weaned at < or = 7 and > 28 d after farrowing were excluded. Farrowing rate excluding sows culled for nonreproductive reasons after mating from the denominator was 85.5%. Parity, farrowing season, and the lactation length x ADFI interaction were associated with farrowing rate (P < .05) using a categorical additive linear model in the CATMOD procedure in SAS (1988). Parity 1 sows had a lower farrowing rate than parities 2 and > or = 6 (P < .05). The summer and spring groups had the lowest and second-lowest farrowing rates among the four seasons (P < .05). In the high ADFI (> or = 5.7 kg) group, no differences in farrowing rate were observed between lactation length groups. In the low (< or = 4.2 kg) and medium (4.2 to 5.6 kg) ADFI groups, farrowing rate varied depending on lactation length. In conclusion, our results suggest that optimizing ADFI (> or = 5.7 kg) alleviates the negative effects of lactation length on farrowing rate. PMID- 9331859 TI - Effect of hemoglobin status on humoral immune response of weanling pigs differing in coping styles. AB - The effects of hemoglobin (Hb) status and coping style of pigs on performance and humoral immune response were studied. Twenty-four, 4-wk-old crossbred barrows were assigned to groups of three pigs based on weight and litter origin. Groups were allotted according to a 2 x 2 factorial treatment arrangement: two blood Hb concentration classes (low vs high) and two immunization procedures (control vs immunized). Immunized pigs received an antigen cocktail containing keyhole limpet hemocyanin (KLH), ovalbumin (OA), and tetanus toxoid (TT) at weaning. Additionally, pigs were stratified according to behavioral coping style in response to exposure to a stressor. During 41 d after weaning (approximate time of immunization), blood Hb concentration, ADG, and ADFI were measured weekly and serum antibody titers to KLH, OA, and TT twice weekly. Average Hb concentration differed between low and high Hb pigs (P < .001; 10.0 vs 12.0 g/dL), but this difference declined with time after weaning. Neither immunization procedure nor coping style affected Hb concentrations. In addition, ADG and ADFI were unaffected by any of the treatments. However, ADG was slightly greater in high Hb status pigs (586 vs 633 g/d) and was paralleled by a slightly greater ADFI in high Hb status pigs (812 vs 899 g/d). Antibody responses were negatively or not related to Hb status at weaning. Antibody responses (depending on isotype and antigen) were or tended to be lower in pigs with high blood Hb concentrations. Behavioral coping style strongly affected humoral immune responsiveness; enhanced or accelerated antibody responses were found in pigs that had a passive coping style. PMID- 9331860 TI - Reproductive performance of cows mated to and preweaning performance of calves sired by Brahman vs alternative subtropically adapted breeds. AB - Comparisons involving Brahman and Brahman-derivative (Brangus, Santa Gertrudis, Beef-master, Simbrah, Braford) sires indicate the following: 1) cows mated to Brangus and Santa Gertrudis bulls had a shorter gestation length than cows mated to Brahman bulls, 2) calves sired by Brangus and Beefmaster bulls were lighter at birth and weaning than calves sired by Brahman bulls, and 3) birth and weaning weights were similar for calves sired by Santa Gertrudis and Brahman bulls and for calves sired by Simbrah and Brahman bulls. Comparisons involving Brahman and other Zebu (Sahiwal, Nellore, Gir, Indu-Brazil, Boran, Romana Red) sires indicate that gestation length was slightly longer for cows mated to Sahiwal and Nellore bulls and that, relative to the Brahman, birth and weaning weights were similar to or lighter for calves sired by bulls of the other Zebu breeds. The only exception to this pattern was birth weight of Indu-Brazil-sired calves, which were heavier than calves sired by Brahman bulls. Comparisons involving Brahman and non-Zebu subtropically adapted (Tuli, Senepol) sires indicate that cows mated to Tuli bulls had a slightly shorter gestation length than cows mated to Brahman bulls and that birth and weaning weights of calves sired by Tuli and Senepol bulls were lighter than those of calves sired by Brahman bulls. PMID- 9331861 TI - Postweaning performance and carcass merit of F1 steers sired by Brahman and alternative subtropically adapted breeds. AB - Comparisons were made among F1 steers sired by Brahman and alternative subtropically adapted breeds of bulls for feedlot and carcass traits when steers were produced from Angus- and Hereford-type dams. Brahman-derivative breeds included Brangus, Beefmaster, and Santa Gertrudis. Brangus- and Beefmaster-sired steers weighed less at slaughter, whereas carcasses of Brangus- and Santa Gertrudis-sired steers had more marbling than those of Brahman-sired steers. Brahman-sired steer carcasses had greater longissimus muscle area than carcasses of Santa Gertrudis-sired steers. Other Zebu breeds compared to Brahman were Boran, Gir, Indu-Brazil, Nellore, Red Brahman, and Sahiwal. Steers by Brahman sires had higher slaughter weights than steers by Boran, Gir, Nellore, or Sahiwal sires. Hot carcass weights of Brahman-sired steers were also higher than those of Boran- and Sahiwal-sired steers. Steer carcasses by Brahman sires had greater longissimus muscle area than those of steers by Sahiwal sires. Non-Zebu breeds included Tuli and Senepol. Steers by Tuli sires grew slower, had lower slaughter weights, and their carcasses weighed less than those of Brahman-sired steers. Brahman-sired steer carcasses had greater longissimus muscle area but less marbling than carcasses of Tuli-sired steers. These data suggest that steers by Brahman sires have an advantage for slaughter weight over steers by Brangus, Beefmaster, Boran, Gir, Nellore, Sahiwal, and Tuli sires, but their carcasses are at a disadvantage for marbling score compared with those by Brangus, Boran, Nellore, and Tuli sires. PMID- 9331862 TI - The effect of prepubertal immunization against gonadotropin-releasing hormone on the development of sexual and social behavior of bulls. AB - To determine the effect of prepubertal immunization against GnRH on the development of sexual and social behavior of Friesian bulls, 90 calves were randomly assigned to five treatments: 1) I2, immunized against GnRH at 2 and boosted at 2.5, 4, and 7.5 mo of age, n = 2 x 10; 2) I4, immunized against GnRH at 4 and boosted at 4.5 and 7.5 mo of age, n = 2 x 10; 3) I7.5, immunized against GnRH at 7.5 and boosted at 8 mo of age n = 2 x 10; 4) S, steers castrated at 2 mo of age, n = 10; and 5) B, intact bulls, n = 2 x 10. Blood samples were collected initially every 2, then every 3 wk. Plasma was analyzed for anti-GnRH titers and plasma testosterone concentration. Sexual and agonistic behavior, male-male mounting, and damage to paddocks was assessed throughout the experiment. All immunized calves developed antibodies against GnRH (32.3 +/- 2.0% bound at a 1:10 plasma:PBS-BSA dilution, 14 d after first boost). Plasma testosterone concentrations were < 1 ng/mL for all immunized animals until 11 mo of age, when they increased to levels found in intact bulls at 14 mo of age. At slaughter, testes and seminal vesicle weights were 38.3 and 31.6% lighter, respectively, for all immunized treatments compared to B. There were no significant differences between I2, I4, and I7.5 in any of the sexual or agonistic behavior tests. Bulls scored higher than steers in all sexual behavior tests. Immunized bulls scored lower than bulls in sexual behavior tests from 10 to 17 mo of age. The proportion of immunized animals that serviced an estrous cow was lower than the proportion of intact bulls at 10, 12.5, 14, and 17 mo of age. Immunized animals scored lower than bulls in bull challenge tests at 8.5, 11.5, 13, 14.5, and 17 mo of age. Paddock damage by animals on the three immunization treatments was lower than that by bulls from 7 to 14.5 mo of age, as were leg were scores (an indicator of male-male mounting behavior) from 9 to 14 mo of age. There was no difference in sexual behavior between immunized bulls (I2, I4, and I7.5) and bulls while held in lairage pens for 16 h before slaughter, but all treatment groups scored higher than steers. There was a similar trend for agonistic behavior, although I4 bulls were no different from steers. Prepubertal immunization against GnRH at 2, 4, and 7.5 mo of age impaired testes function and affected the development of social and sexual behavior of young bulls. PMID- 9331863 TI - Anabolic implant effects on visceral organ mass, chemical body composition, and estimated energetic efficiency in cloned (genetically identical) beef steers. AB - Six sets of four genetically identical Brangus steers (n = 24; X BW 409 kg) were used to determine the effect of different anabolic implants on visceral organ mass, chemical body composition, estimated tissue deposition, and energetic efficiency. Steers within a clone set were randomly assigned to one of the following implant treatments: C, no implant; E, estrogenic; A, androgenic, or AE, androgenic + estrogenic. Steers were slaughtered 112 d after implanting; visceral organs were weighed and final body composition determined by mechanical grinding and chemical analysis of the empty body. Mass of the empty gastrointestinal tract (GIT) was reduced approximately 9% (P < .10) in steers implanted with estrogen alone or in combination with an androgen. Liver mass was increased (P < .10) from 6 to 14% by implants. Steers implanted with the AE combination had greater (P < .10) daily protein accretion (163.4 g/d) than either E (128.8 g/d) or A (137.1 g/d), and, because the combination improved gain above C (101.1 g/d), this demonstrates the additive effects of a combination implant on protein deposition. Anabolic implants did not alter (P > .10) the efficiency of ME utilization. In general, estrogenic implants decreased GIT, androgenic implants increased liver, and all implants increased hide mass. Steers implanted with an AE combination had additive effects on protein deposition compared with either implant alone. The NEg requirements for body gain are estimated to be reduced 19% by estrogenic or combination implants. PMID- 9331865 TI - Vitamin E supplementation of cattle and shelf-life of beef for the Japanese market. AB - Feeder steers (n = 84) were stratified into four weight groups to provide slaughter groups so that product that had been in vacuum packages at 0 to 2 degrees C for 40, 60, 80, or 100 d postmortem could be simultaneously evaluated. Each of the four groups was randomly divided into three subgroups so that vitamin E could be supplemented in the diet at rates of 0, 1,000, or 2,000 (E0, E1000, and E2000, respectively) IU.steer-1.d-1 for 100 d. After slaughtering, chilling, and fabricating, one ribeye-roll and one strip loin from each carcass was transported to the university laboratory for analyses, whereas the paired subprimals were transported to Japan. Based on metmyoglobin formation and lipid oxidation, strip loin steaks deteriorated at a faster rate during retail-display than did ribeye steaks. Steaks from subprimals that were stored for 100 d had inferior (P < .05) retail-display characteristics and a shorter (P < .05) caselife than steaks from the other storage periods. alpha-Tocopherol levels in longissimus muscle were lower (P < .05) for E0 than for E1000 and E2000 (3.51, 5.54, and 6.10 micrograms/g of tissue, respectively). Supplementing cattle with vitamin E resulted in steaks that exhibited superior lean color, less surface discoloration, more desirable overall appearance, and less lipid oxidation during retail display than control steaks; minimal differences were observed between E1000 and E2000 steaks. Steaks from cattle supplemented with vitamin E were preferred over control steaks by 91% of Japanese survey participants (n = 10,941), and 58% of all participants identified muscle color as the most important factor in selecting beef products. PMID- 9331864 TI - Serum concentrations of trenbolone-17 beta and estradiol-17 beta and performance of heifers treated with trenbolone acetate, melengestrol acetate, or estradiol-17 beta. AB - The effects of the growth-promoting steroids estradiol-17 beta (E2), trenbolone acetate (TBA), and melengestrol acetate (MGA) in heifers on serum concentrations of E2 and trenbolone-17 beta (TBOH) were examined. Feed intake and growth performance were also measured. Serum concentrations of E2 and TBOH were measured on d 0, 1, 3, 5, 7, 13, 21, 28, 42, 56, 84, 112, and 140 in finishing heifers administered the following treatments: 1) control; 2) MGA, .5 mg per heifer daily; 3) Revalor-H (140 mg TBA + 14 mg E2); 4) Revalor-H + MGA; 5) Finaplix-H (200 mg TBA); and 6) Finaplix-H + MGA. Revalor-H implantation (Treatments 3 and 4) increased (P < .05) serum E2 concentrations; peak concentrations (67.5 pg/mL) occurred between d 21 and 56. Feeding MGA (Treatment 4) had no effect (P > .05) on this increase in serum E2 concentrations (63.3 pg/mL). From d 84 until d 140, serum E2 was greater (P < .05) for the Revalor-H treatment (average of 19 pg/mL) than for the control (7 pg/mL) or Finaplix-H treatments (6.5 pg/mL). Serum E2 concentrations increased numerically two- to threefold from d 56 to 140 in controls fed MGA, compared with controls not fed MGA. There was the expected increase in serum TBOH concentrations after TBA implantation in the Revalor-H and Finaplix-H treatments; concentrations were similar (P > .05) for Revalor-H (221 pg/mL) and Finaplix-H (280 pg/mL). After d 56, serum TBOH concentrations decreased in both treatments to 10 and 20% of these concentrations, respectively. Feeding MGA increased serum TBOH (P < .05). Dry matter intake by heifers did not differ among treatments. Feeding of MGA improved gains (P = .12) and efficiencies (P < .01) in nonimplanted heifers and had no effect (P > .4) on gains or efficiencies in Finaplix-implanted heifers. PMID- 9331866 TI - Sensory characteristics and carcass traits of boars, barrows, and gilts fed high- or adequate-protein diets and slaughtered at 100 or 110 kilograms. AB - The objective of this study was to determine consumer reaction to boar (BO), barrow (BA), and gilt (G) meat from pigs grown and finished on high- (HI) and low (LO) protein diets and slaughtered at 100 and 110 kg BW. Within each of two trials, 54 BO, BA, and G were allotted within sexes to HI or LO protein sequences for growing and finishing: 19 and 17% (BOHI), 18 and 16% (BOLO), 17 and 15% (GHI), 16 and 14% (GLO), 15 and 13% (BAHI), and 14 and 12% (BALO). Backfat skatole and salivary gland 16-androstene concentrations were measured from samples taken at slaughter. Longissimus (LM) and semitendinosus (ST) chops from 24 pigs (with equal representation across diet and sex groups) were evaluated by trained panelists for tenderness, juiciness, and off-flavor. Consumer panelists evaluated acceptability of LM chops. In the 100-kg trial, HI diets improved (P < .05) carcass leanness in BO and BA but not in G. In both trials, BO were leaner (P < .05) than G, and both were leaner (P < .05) than BA. Skatole and 16 androstene concentrations were similar (P > .05) among sexes in both trials. In the 100-kg trial, trained panelists found BOLO chops had more (P < .05) off flavor. In the 110-kg trial, all BO had more off-flavor (P < .05) than BAHI, BALO, and GHI but were similar (P > .05) to GLO. In both trials, BA chops were more tender (P < .05) than G and BO chops and LM chops had less off-flavor (P < .05) than ST chops. In the 110-kg trial, skatole was correlated (r = .28, P < .001) to off-flavor. A relationship may exist among diet, skatole deposition, and off-flavor. Untrained consumers reported all chops were equally acceptable (P > .05). PMID- 9331867 TI - Effect of postmortem injection time and postinjection aging time on the calcium activated tenderization process in beef. AB - The objectives of this study were to determine 1) the effectiveness of calcium chloride when injected later than 2 d postmortem, 2) the effect of extended postinjection aging time, and 3) the tenderness response curve in calcium chloride-treated beef. In Exp. 1, the longissimus thoracis et lumborum was injected on either d 2 or 14 postmortem with 5% by weight of a 200 mM calcium chloride solution. Samples were aged (1 degree C) either 7 or 35 d after injection. The uninjected control longissimus from the contralateral side was aged for 9, 21, 37, or 49 d. In Exp. 2, the longissimus thoracis et lumborum was injected on d 2 postmortem with 5% by weight of a 200 m x M calcium chloride solution then sampled for shear force on d 1, 2, 6, 8, 12, and 14 after injection. Calcium chloride injection, regardless of injection time or postinjection aging time, had higher (P < .05) sensory tenderness rating than control with the same total aging time (5.2, 5.5, 5.8, and 6.1 vs 4.3, 4.8, 5.1, and 5.3, respectively). Calcium chloride injection at d 14 reduced shear force (.7 kg) and increased tenderness rating (.7 units) as effectively (P > .05) as injection at d 2 (1.2 kg and .8 units, respectively). Calcium chloride-injected steaks had higher (P < .05) juiciness ratings than control steaks. Postrigor calcium chloride injection reduced (P < .05) shear force within 1 d after injection and resulted in more tender meat through 14 d after injection. Extended postinjection aging (35 d) had little effect on color display stability. Calcium activated tenderization can be applied as late as 14 d postmortem and will reduce the occurrence of tough meat if aging is limited. PMID- 9331868 TI - Efficacy of chromium picolinate and chromium chloride as potential carcass modifiers in swine. AB - We conducted two experiments to evaluate the effects of chromium picolinate and chromium chloride (CrCl3) on growth performance, carcass composition, percentages and accretion rates of carcass tissues and chemical components, and blood metabolites in pigs. In Exp. 1, 35 individually penned pigs were fed a fortified, corn-soybean meal basal diet (.95% lysine) supplemented with 0, 200, or 400 micrograms/kg of Cr from chromium picolinate or 5,000 or 25,000 micrograms/ kg of Cr from CrCl3. Each diet was fed to seven pigs for 35 d (19.6 to 43.2 kg BW). Addition of 200 micrograms/kg of Cr from chromium picolinate increased ADG (P < .07) and ADFI (P < .03) but did not affect feed:gain ratio. Backfat measurements and longissimus muscle area were not affected by either source of Cr. The percentages of muscle, fat, bone, and skin from the right ham and the percentages of water, protein, lipid, and ash from the left carcass were not significantly altered by Cr. The addition of 200 micrograms/kg Cr from chromium picolinate increased (P < .07) the accretion rate of lipid in the carcass. In Exp. 2, 42 individually penned pigs (three from each of 14 litters) were fed a fortified, corn-soybean meal basal diet (.95% lysine from 19 to 55 kg; .80% lysine from 55 to 109 kg) without or with 200 micrograms/kg of Cr from chromium picolinate or 5,000 micrograms/kg of Cr from CrCl3. Dietary Cr addition had no effect on the performance or backfat measurements of the pigs; however, both sources of Cr increased (P < .07) longissimus muscle area. The percentages and accretion rates of muscle tissue were increased (P < .001) and the percentages of fat tissue were decreased (P < .001) in pigs fed Cr, with chromium picolinate being more effective than CrCl3 (P < .05). The percentages (P < .01) and accretion rates (P < .07) of carcass protein were increased and the percentages and accretion rates of carcass lipid were decreased (P < .04) in pigs fed Cr. No changes in blood metabolites occurred as a result of supplemental Cr in either experiment. These results suggest that chromium picolinate is more effective than CrCl3 and that Cr must be supplemented throughout the growing-finishing period to improve the carcass composition. PMID- 9331869 TI - Estimation of relative bioavailability of nutrients using SAS procedures. AB - The General Linear Models procedure (PROC GLM) in SAS/STAT software can be programmed to perform the standard statistical analyses used for relative bioavailability studies. The first steps are validity checks to test for statistical validity (linearity), fundamental validity (intersection of regression lines at 0 supplemental level), and equality of the basal diet mean to the point of intersection. The CLASS variable capabilities of PROC GLM can be exploited to expedite these tests. After the validity checks, the GLM procedure can be used to obtain parameter estimates for calculation of relative bioavailability. Optional output provides an inverse matrix to calculate standard errors of slopes and slope ratios. Logarithmic and other transformations of the dependent variable to reduce variance heterogeneity or achieve linearity for subsequent calculation of appropriate bioavailability values also can be accomplished within the SAS System. When nonlinear regression models are more appropriate than linear models, the NLIN procedure can be used. PMID- 9331870 TI - Vitamin A repletion in thoroughbred mares with retinyl palmitate or beta carotene. AB - Forty-five Thoroughbred mares used in an 8-mo depletion study were kept for an additional 20 mo on the same three forage diets (15 mares each): 2-yr-old orchardgrass hay and vitamin A-free concentrate on a drylot (HC); pasture, orchardgrass/alfalfa hay, and vitamin A-free concentrate (PHC); or pasture and orchardgrass/alfalfa hay only (PH). Each diet group was divided into three subgroups, and mares (n = 5) in each group were given either retinyl palmitate (A) at twice the NRC (1989) recommended daily intake, the equivalent amount of vitamin A in the form of water-dispersible beta-carotene (B), or the vehicle (C). Vitamin A status was monitored with serum retinol and a relative dose response (RDR) test every 60 d. In the C subgroups, retinol concentration was 18.65 +/- .84 micrograms/dL (mean +/- SE) and the RDR was 16.26 +/- 1.72% over the 20 mo. Retinol and RDR fluctuated seasonally regardless of supplementation. Vitamin A status, based on serum retinol (P = .001) and RDR (P < .001) values, was lower in the HC than in the PH and PHC. Vitamin A status, based on retinol (P = .05) and RDR (P = .013) values, was improved by retinyl palmitate supplementation in all diet groups, but not by water-dispersible beta-carotene supplementation. Supplementation of the HC mares with vitamin A matched the serum retinol, but not the RDR, of the two pasture, control subgroups. Thus, replete vitamin A status in previously depleted mares was barely obtained by supplementation with twice the currently recommended daily intake of vitamin A. PMID- 9331871 TI - Effectiveness of a pair-gain feeding strategy for individually fed, group-housed finishing pigs. AB - Crossbred barrows (n = 48) and gilts (n = 12) were used to examine the effectiveness of a pair-gain feeding strategy for individually fed, group-housed barrows. In a pair-gain feeding strategy, barrows were individually restricted to a feeding level at which their growth was similar to the mean growth of gilts with ad libitum access to feed. The purposes of this feeding strategy were to have barrows and gilts reach slaughter weight at the same time and to improve carcass traits of the barrows. At 29.8 +/- .4 kg BW, barrows were assigned to either the pair-gain or the ad libitum treatment. All pigs had free access to feed until they reached 60 kg BW. The experimental period was from 60 to 110 kg BW. The 12 group-fed gilts and 24 individually fed barrows (12 per pen) were also given free access to feed throughout the experimental period. The remaining 24 barrows (12 per pen) were put on a pair-gain feeding strategy. In the pair-gain feeding strategy, the weekly feed allowance of each barrow was based on its measured BW and computed energy conversion ratio and on the mean growth of the gilts. The barrows in the pair-gain treatment grew at the same rate as gilts. The ad libitum intake barrows grew faster (P < .05) and had a poorer energy conversion ratio for production than the barrows in the pair-gain treatment. The total energy conversion ratio, backfat thickness, and lean meat percentage were similar (P > .10) for the two treatments. In conclusion, the pair-gain feeding strategy was effective in achieving similar growth between barrows and gilts. The total energy conversion ratio and carcass traits of the barrows, however, were not improved. PMID- 9331873 TI - Swine nutrition: nutrient usage during pregnancy and early postnatal growth, an introduction. AB - This symposium was organized to review current knowledge of the nutrient requirements of sows during pregnancy and of pigs soon after birth. It consisted of five presentations that focused on energy metabolism, fiber as an energy source, protein requirements, vitamin and mineral requirements, and the special needs of newborn pigs. Additionally, this symposium was organized to honor the immense contributions of Dr. Wilson Pond to this field. The purpose of the introduction to this symposium was to highlight the major areas of research that were discussed within each presentation and the work of Dr. Pond that the present research uses as a foundation. PMID- 9331872 TI - Effect of resistant starch on net portal-drained viscera flux of glucose, volatile fatty acids, urea, and ammonia in growing pigs. AB - Net portal-drained viscera (PDV) flux of glucose, VFA, ammonia, and urea was determined in pigs fed diets with or without resistant starch. Diets consisted of 65% cornstarch (diet CS), 32.5% cornstarch and 32.5% raw potato starch (diet CPS), or 65% raw potato starch (diet PS); the remaining 35% supplied all amino acids, fat, fiber, minerals, and vitamins. The diets contained twice the maintenance requirement for energy and were fed twice daily to four barrows (initial BW 56 kg) in three periods in a crossover design. The pigs were fitted with catheters in a mesenteric vein, a mesenteric-artery, and the portal vein, and net PDV flux was calculated by multiplying portal-arterial concentration differences and corresponding portal vein flow. Net PDV flux of glucose was significantly less after feeding diets CPS and PS, and portal absorption of ileally digested glucose was 89, 66, and 41% for diets CS, CPS, and PS, respectively. Net PDV flux of VFA was lowest after feeding diet CS and three to four times higher after feeding diets CPS and PS. Net PDV flux of ammonia was highest for diet CS and almost halved after feeding diets CPS and PS. There was a small negative net PDV flux of urea for diets CS and CPS, which significantly increased after feeding diet PS. These results suggest that excretion of nitrogen is shifted from urine to feces primarily by reduction of the net PDV flux of ammonia when resistant starch is fed. PMID- 9331874 TI - Energy metabolism in pregnant sows and newborn pigs. AB - Measurements of heat production(HP; indirect calorimetry) and its partition between maintenance, physical activity, thermoregulation, and thermic effect of feed or energy gain were carried out in sows maintained in different situations: primiparous or multiparous; pregnant or nonpregnant; thermoneutral or cold conditions; varied feeding levels; and varied body weights (BW). Metabolizable energy requirements for maintenance average 420 kJ/kg BW.75 at thermoneutrality and moderate physical activity. This value is not significantly affected by parity, pregnancy, and stage of pregnancy. Physical activity is a major factor causing differences in energy balance between sows because activity is variable and its energy cost (27 kJ.kg BW-.75.100 min-1 standing) is four to five times higher than in other species. Lower critical temperature (LCT) is approximately 20 degrees C in pregnant and individually housed sows, and daily HP is increased by approximately 15 kJ/kg BW.75 for each degree Celsius decrease of ambient temperature below LCT. Efficiencies of utilization of ME for meeting energy requirements for maintenance, maternal gain, and uterine gain are 77, 75, and 50%, respectively. Equations for predicting energy deposition in the uterus and mammary gland are proposed. In addition to activity and thermoregulation, energy requirements of pregnant sows depend on body reserves of energy. Studies with newborn pigs indicate that they are quite sensitive to ambient temperature (i.e., +25 J.kg BW-.75.min-1 for each degree Celsius decrease of temperature), and their LCT is 32 to 34 degrees C. The energy demand of pigs for thermoregulation just after birth relies mainly on carbohydrates from glycogen reserves or colostrum. Survival of newborn pigs is highly dependent on the supply of colostrum. PMID- 9331875 TI - Microbial perspective on fiber utilization by swine. AB - Dietary fiber may contribute up to 30% of the maintenance energy needs of growing pigs. Higher energy contributions may be obtained from dietary fiber fed to sows, along with some improvements in reproduction, health, and well-being. As long as cereal grain supplies and high-quality protein supplements are abundant, the use of fibrous feeds for swine most likely will be limited. However, as the human demand for cereal grains increases, swine producers, especially those with reproductive animals, may be economically forced to incorporate alternative feedstuffs. These feedstuffs might include lignified plant cell wall material such as grasses and legumes, and feed-milling and distillery by-products that contain a high level of fiber residues. The microflora in swine large intestine will be able to adapt to these lignified forages and by-product feeds much better than the microflora in humans. Swine microflora contain highly active ruminal cellulolytic and hemicellulolytic bacterial species, which include Fibrobacter succinogenes (intestinalis), Ruminococcus albus, Ruminococcus flavefaciens, Butyrivibrio spp., and Prevotella ruminicola. Additionally, a new highly active cellulolytic bacterium, Clostridium herbivorans, has been recently isolated from pig large intestine. The populations of these microorganisms are known to increase in response to the ingestion of diets high in plant cell wall material. The numbers of cellulolytic bacteria from adult animals are approximately 6.7 times greater than those found in growing pigs. None of these highly active cellulolytic bacterial species are found in the human large intestine. Thus, the pig large intestinal fermentation of fiber seems to more closely resemble that of ruminants than that of humans. PMID- 9331876 TI - Protein nutrition of gestating sows. AB - Dietary amino acid requirements of gestating sows vary depending on genetic strain, energy intake, body weight, and other factors. Estimation of amino acid requirements for gestating sows in a defined situation should be directed toward achieving a high rate of protein accretion and a modest rate of fat accretion. More information is needed to set target accretion rates precisely. A simple mathematical model was developed with the limited objectives of 1) aiding in understanding the interactions among energy intake, amino acid intake, protein accretion potential, and body weight in affecting the amount of body protein and fat accreted by pregnant sows and 2) providing general guidelines for feeding programs. Predictions from the model suggest that the daily lysine requirement varies markedly with variation in ME intake but that the requirement as a percentage of the diet is more stable as ME intake varies. At moderate ME intake, the lysine requirement seems greater during late gestation than earlier. The lysine requirement of heavy sows is less than that of light gilts at a similar ME intake, and this offers opportunity for modest cost savings in pork production. A compromise is necessary between the goals of a high rate of protein accretion and a modest rate of fat accretion. It seems that a sow's potential protein accretion rate is rarely limiting. PMID- 9331877 TI - Vitamin and mineral transfer during fetal development and the early postnatal period in pigs. AB - There are periods during pregnancy when sows may have a temporally high requirement for certain vitamins and minerals. Proteins transferring retinol and Fe to the developing pig fetus have been discovered, whereas transport mechanisms for other vitamins and minerals are probably present but have not yet been identified. Sow body tissues can serve as a reservoir for many micronutrients, but it is not known whether these reserves can supply an adequate quantity during critical fetal developmental periods. There is a low placental transfer of vitamin E to the fetus even if the dietary concentration fed to a gestating animal is high, but colostrum and milk concentrations can be increased when the nutrient is fed to sows. If the dam's diet contains inadequate Ca or P, the concentration of these elements in the developing fetus and milk will not be affected. Consequently, sow bone demineralization will occur under conditions of dietary inadequacy of Ca and P. Other nutrients can be depleted from sow tissue reservoirs over several parities (e.g., Se), resulting in low quantities being provided in the milk for nursing pigs. Scientific information involving adequate vitamin and mineral nutrition for female pigs to improve conception rate and embryonal survival that will result in optimum fetal and postnatal pig development can be considered to be in its infancy. PMID- 9331878 TI - Role of milk-borne vs endogenous insulin-like growth factor I in neonatal growth. AB - Neonatal pigs are characterized by a high efficiency of nutrient utilization and rapid growth rate. The utilization of dietary protein for lean tissue growth is particularly efficient in neonatal pigs and is associated with a high rate of skeletal muscle protein synthesis and deposition. In support of these high growth rates, neonatal pigs consume a milk diet that has a high biological value and is abundant in growth factors, including insulin and IGF-I. During the neonatal period, there are developmental changes in the circulating concentrations of, and tissue responsiveness to, hormones, particularly insulin, IGF-I, and growth hormone that play a central role in growth regulation. Our goal has been to characterize the dietary factors and specific aspects of endocrine function that are responsible for the anabolic stimulus that helps to sustain the high rates of protein deposition in neonatal pigs. Our results suggest that, despite the abundance of growth factors in milk and colostrum, the intake of nutrients is the primary anabolic stimulus for protein synthesis and this response declines with age. There is, however, a nonnutritive and as-yet-unidentified component in colostrum that provides a specific anabolic stimulus for skeletal muscle in newborns, but this is probably neither insulin nor IGF-I. Our studies also indicate that circulating concentration of IGF-I are not a primary stimulus of skeletal muscle protein synthesis and that the primary endocrine signal that mediates the response to nutrient intake may be insulin. Future research should address how the local expression of IGF and the function of insulin and IGF receptors affect the responsiveness of anabolic processes to nutrient intake and hence the efficiency of neonatal growth. PMID- 9331879 TI - Mechanisms of action of growth hormone-releasing peptide-2 in bovine pituitary cells. AB - We conducted this study to investigate the mechanisms of action of growth hormone releasing peptide-2 (D-Ala-D-beta Nal-Ala-Trp-D-Phe-Lys-NH2; GHRP-2) in bovine anterior pituitary primary cell culture. Doses of GHRP-2 from 10(-13) to 10(-7) M) increased (P < .05) GH secretion. The GHRP-2 (10(-7) M) and GH-releasing factor (GRF; 10(-7) M) administered together had an additive effect on the release of GH (P < .05). Somatostatin (1 microM) decreased GH secretion in response to GHRP-2 and(or) GRF (P < .05). Secretion of GH in response to GHRP-2 was blocked (P < .01) by a GRF receptor antagonist (.1 microM). Nifedipine (10 microM), a voltage-dependent Ca2+ channel blocker, inhibited (P < .01) GHRP-2 stimulated GH release. The GH release in response to GHRP-2 and 4 beta-phorbol-12 myristate-13-acetate (10(-7) M), a protein kinase C activator, was additive (P < .01). Forskolin (30 microM), a cAMP elevating agent, further stimulated (P < .01) the GH release in response to GHRP-2. Bovine GH concentrations in culture media were assayed by indirect competitive enzyme immunoassay. These results showed that GHRP-2 1) stimulates GH secretion from bovine pituitary cells, 2) may partially act via GRF receptor, 3) has GH secretion activity caused by Ca2+ influx via Ca2+ channels, and 4) may increase GH secretion via protein kinase C and cAMP pathways. PMID- 9331880 TI - No desensitization of the growth hormone (GH) response between GH-releasing peptide-2 and GH-releasing factor in calves. AB - We investigated the desensitization caused by growth hormone (GH)-releasing peptide-2 (D-Ala-D-beta Nal-Ala-Trp-D-Phe-Lys-NH2; GHRP-2) in vivo using calves. The GHRP-2 (12.5 micrograms.kgBW-1.h-1) or GH-releasing factor (GRF; .125 microgram.kgBW-1.h-1) were infused for 180 min into four calves, each followed by either GHRP-2 (12.5 micrograms/kg BW) or GRF (.125 microgram/kg BW) injection after 60 min. All treatments were given to all of the calves. The continuous infusion of GHRP-2 did not attenuate the GH secretion caused by a GRF bolus, and vice versa. Continuous exposure to GHRP-2 attenuated the GH secretion caused by a GHRP-2 bolus. In an additional experiment, two repetitive injections of GHRP-2 (12.5 micrograms/kg BW) at 1-, 2-, 3-, or 4-h intervals were administered. Attenuation of the GH response caused by GHRP-2 was maintained until 4 h. These results suggest that GHRP-2 is not cross-desensitized with GRF and that the mechanisms by which they cause GH release may be different in calves. PMID- 9331881 TI - Luteinizing hormone release and reproductive traits in anestrous, estrus-cycling, and ovariectomized cattle after tyrosine supplementation. AB - The role of rumen-protected L-tyrosine as a nutritional signal altering LH release and other reproductive traits in cattle was studied. In Exp. 1, 28 suckled crossbred cows were assigned randomly to five treatments (0 or 40 g of tyrosine daily in feed for 3 d followed by a single i.v. injection of 200 micrograms of GnRH or 1 mg/kg of naloxone on d 26 +/- 1 postpartum; no tyrosine plus an injection of saline was the control). Peak LH after GnRH was greater (P < .001) in GnRH-treated cows regardless of tyrosine supplementation. Compared to cows receiving saline, days to first postpartum ovulation were reduced (P < .05) by naloxone, tyrosine, and GnRH. In Exp. 2, 47 suckled crossbred cows were assigned randomly to six treatments (0, 20, or 40 g of tyrosine daily for 3 d before GnRH or saline was injected i.m. on d 23 +/- 1 postpartum). Injection of GnRH increased (P < .001) LH. An interaction (P = .08) of tyrosine and GnRH tended to reduce days to first postpartum ovulation. In Exp. 3, tyrosine (40 g) administered once daily for 3 d to ovariectomized cows (six cows per treatment) had no effect on any characteristic of LH before or after estradiol-17 beta. In Exp. 4, suckled cows (n = 136) were allotted randomly to two treatments (0 or 30 g of tyrosine daily for 3 d before a PGF2 alpha-synchronized estrus). Tyrosine increased (P = .05) the percentage of cows in estrus after PGF2 alpha but reduced (P = .05) AI conception rate. These results fail to support the thesis that tyrosine alters LH release in cattle. Supplemental tyrosine increased expression of estrus in suckled cows after PGF2 alpha and tended to reduce intervals to first postpartum ovulation. PMID- 9331882 TI - Ad libitum suckling by an unrelated calf in the presence or absence of a cow's own calf prolongs postpartum anovulation. AB - Our objective was to determine whether onset of first postpartum ovulation would be altered in suckled cows nursing an unrelated calf in the presence or absence of their own nonsuckling calf. In a 2-yr study, Angus x Hereford cows were assigned randomly to five treatments between d 13 and 18 postpartum for 4 wk: 1) own calf was weaned (OCW, n = 9); 2) own calf was present continuously (OCP, n = 12); 3) own calf was present continuously but restricted from udder contact (OCR, n = 9); 4) unrelated calf was present continuously after own calf was removed (UCP, n = 10); and 5) unrelated calf was present continuously and own calf was present but restricted as in the OCR treatment (OCR + UCP, n = 10). Interval to the first increase in progesterone (ovulation) was less (P < .05) in OCW (14.7 +/ 3.4 d) and OCR (19.9 +/- 3.4 d) than in the OCP (35.0 +/- 2.9 d), UCP (38.0 +/- 3.4 d), and OCR + UCP (37.6 +/- 3.4 d) treatments. The OCW cows showed no maternal bond with their own calves after 4 wk of treatment, whereas OCR, OCP, and OCR + UCP cows were bonded to their own natural calves. Cows in the UCP treatment, suckled by unrelated calves, formed new maternal bonds with those calves, resulting in prolonged anovulation. Further, OCR + UCP cows had prolonged anovulation because maternal bonds were maintained with their own restricted calves while milk was removed by unrelated calves. We conclude that milk removal preceded by a continuously reinforced cow-calf bond (original or reestablished) is essential to prolong anovulation in beef cows. PMID- 9331883 TI - Pulsatile release of somatotropin related to meal feeding and somatotropin response to secretagogues in horses. AB - Our goal was to establish a time of day and(or) interval from feeding that would avoid the refractory period after a somatotropin (ST) surge and optimize the responsiveness of horses to ST secretagogues. Two experiments were conducted with eight geldings conditioned to consume grain at 0800 and 1600 daily. In Exp. 1, during a 24-h period, these geldings averaged 3.2 +/- .3 pulses of ST with peak amplitude of 4.2 +/- .4 ng/mL, pulse duration of 55 +/- 6 min, and interpeak interval of 400 +/- 57 min. No ST peaks occurred within 2 h after either grain feeding. In Exp. 2, eight geldings were given 50 micrograms of ST-releasing factor (STRF) at 0800. Two geldings that had a pulse of ST between 0700 and 0800 failed to respond to STRF, but the other six responded with a pulse of ST at 37 +/- 3 min; peak amplitude was 4.6 +/- 2.2 ng/mL and duration was 123 +/- 25 min. Experiments 3 and 4 were with mares aged 20 to 26 yr and conditioned to be fed grain at 0800 daily. In Exp. 3, blood was sampled for 8 h beginning at 0500. Seven of the eight mares had a ST pulse in progress at 0500. Five additional pulses were detected, all from 0740 to 0940, but none from 0600 to 0700 or from 1000 to 1300. In Exp. 4, four of the same eight mares were given 50 micrograms of STRF at 0700 and the other four at 1300. Three of the four treated at 0700 and all four treated at 1300 responded to STRF with ST peaks at 20 +/- 5 min; peak amplitude was 12.7 +/- 9.5 ng/mL and duration was 69 +/- 6 min. In Exp. 5, nine mares aged 20 to 26 yr were fed grain at 0800 and 1600 as in Exp. 1 and 2 and given a nonpeptidal ST secretagogue (STS, Merck L-163,255) i.v. at 0, 1, or 5 mg/kg (n = 3 mares/dose) at 1300. No mare had a pulse of ST during the 1 h before treatment. All six mares given STS responded with ST pulses. The ST responses to STS at 1 and 5 mg/kg did not differ (P > .05); time to ST peak was 35 +/- 4 min, pulse amplitude was 24.0 +/- 6.3 ng/mL, and pulse duration was 100 +/- 9 min. We conclude that mares and geldings fed grain once or twice daily usually have a period of 2 to 5 h after feeding with no ST pulses. When horses are fed grain at 0800, one may give a ST secretagogue at 1300 to avoid a refractory period and improve the probability of an ST response. PMID- 9331884 TI - Estradiol-17 beta and progesterone increase ovine uterine suppressor cell activity. AB - We evaluated the regulation of ovine uterine (UT) suppressor cell activity by progesterone (P4), estradiol-17 beta (E2), and P4 + E2 in ovariectomized (OVX) ewes. Following 14 d of steroid injections, endometrial cells (designated as UT cells) were recovered postmortem, and unfractionated and fractionated cells were assessed for suppression of autologous phytohemagglutinin (PHA)-treated peripheral blood lymphocytes (PBL). Supernatants from cultured UT cells were also assessed for suppressor activity. In other experiments, UT cells recovered from nontreated OVX ewes were cocultured with PHA-treated PBL and varying concentrations (1 x 10(-11) to 1 x 10(-5) M) of each steroid preparation. Supernatants from separate cultures that contained UT cells and steroids were evaluated for suppressor activity. Uterine cells from control and steroid-treated ewes suppressed proliferative responses of PHA-treated PBL; however, suppressor activity of UT cells was greater (P < .05) for E2-treated than for control and P4 treated ewes. Uterine suppressor cells from steroid-treated ewes sedimented in Percoll within a density range of 1.002 to 1.056 g/mL. Uterine cells from all ewes released suppressor factor(s) into the culture medium; however, the activity of the supernatant from the cultured cells was not increased for the steroid treated ewes. For cocultures that contained steroids and cultures that contained supernatant, suppressor activity of the UT cells was increased by specific concentrations of each steroid preparation. These findings demonstrate that reproductive steroids augment ovine UT suppressor cell activity. PMID- 9331885 TI - Ruminal undegradability of blood meal and effects of blood meal on ruminal and postruminal digestion in steers consuming vegetative orchardgrass hay. AB - Four crossbred steers (360 +/- 3 kg) cannulated at the rumen and duodenum were used in a 4 x 4 Latin square design to examine the effects of supplemental blood meal (BM) on voluntary intake, digesta kinetics, ruminal fermentation, site and extent of digestion, and bacterial protein synthesis in steers fed vegetative orchardgrass hay (Dactylis glomerata L.). The levels of BM supplementation were 0, .07, .13, and .20 kg/d. Voluntary intake of OM (8.35 kg/d) was not significantly affected by BM supplementation. No significant effects of BM supplementation were detected for OM flow to the duodenum or digestion in the rumen or lower tract. However, total tract OM digestibility decreased 2.2 percentage units at .20 kg/d of BM intake (lower with vs without BM; P < .10). Total N intake and flow to the duodenum linearly increased (P < .10) with increasing BM level from 251 to 277 g/d and from 158 to 199 g/d, respectively. Ammonia N and bacterial N flows to the duodenum were not affected (P > .10) by BM supplementation. As a result, nonammonia N flow to the duodenum increased linearly (P < .10) with increasing BM supplementation. Ruminal escape N from BM was 83.5, 85.3, and 87.2% for .07, .13, and .20 kg/d of BM, respectively. Apparent bacterial efficiency and true bacterial efficiency were not affected (P > .10) by BM supplementation. Total amino acid and total essential amino acid flows to the duodenum were increased (P < .10) by dietary inclusion of BM. Duodenal flows of all essential amino acids except lysine and valine and of all nonessential amino acids except alanine and proline were increased (P < .10) by BM inclusion in the diet. In summary, supplementation with BM increased ruminal escape N and duodenal flows of total and most essential amino acids. PMID- 9331886 TI - Escape protein supplementation of steers fed grass silage-based diets. AB - Beef steers (Trials 1 and 2, 280 +/- 2 kg; Trial 3, 330 +/- 2 kg) were fed diets of 67.5% wheat (Trial 1) or oat silage (Trials 2 and 3), 20.5% barley, and 12% supplement in randomized complete block design growth trials. Dietary treatments were graded levels of supplemental escape protein (EP) from corn gluten meal (Trial 1, 0 to 213 g/d escape protein) or animal by-products (1:1:1 DM mixture of blood, feather, and meat and bone meals, 0 to 223 or 0 to 317 g/d of escape protein in Trials 2 and 3, respectively) to titrate amounts needed to maximize steer live weight gain. As supplemental EP from corn gluten meal increased, steer live weight gain increased linearly (P < .001) and feed-to-gain decreased linearly (P < .001). Supplementation with 135 g/d of corn gluten meal EP (335 g/d of corn gluten meal) increased average daily gain from .76 to .91 kg/d. As supplemental EP from animal by-products increased, steer live weight gain increased quadratically (P < .05) and feed-to-gain decreased linearly (P < .01). Supplementation with 223 or 317 g/d of animal by-product EP increased live weight gain by .27 kg/d. Supplemental escape protein was needed to increase live weight gain of steers consuming ensiled forage diets due to low EP contents of silages (7, 3, and 23% of CP in Trials 1, 2, and 3, respectively) and barley (15, 27, and 22% of CP in Trials 1, 2, and 3, respectively) and limited microbial protein synthesis. PMID- 9331887 TI - Effects of saturation and esterification of fat sources on site and extent of digestion in steers: ruminal fermentation and digestion of organic matter, fiber, and nitrogen. AB - Five steers (average 526 kg) fitted with ruminal, duodenal, and ileal cannulas were used in a 5 x 6 Youden square design with 14-d periods. Diets contained chopped alfalfa hay, corn silage, and concentrate (25:35:40, DM basis). Treatments were 1) control (no added fat), 2) tallow (T), 3) partially hydrogenated tallow (PHT), 4) hydrogenated tallow (HT), 5) blend (1:1) of HT and hydrogenated free fatty acids (HTHFA), and 6) hydrogenated free fatty acids (HFA). Fats replaced cornstarch in the control diet to supply 5% added fatty acids. Intake was restricted to 90% of ad libitum; DMI was similar among diets (average 9 kg/d). Ruminal pH and molar proportion of propionate (P) were greater (P < .05) but total VFA concentration, proportion of acetate (A), A:P, and percentages of OM digested in the rumen and total tract were less (P < .05) when fat-supplemented diets were fed than when the control diet was fed. Total VFA concentration increased linearly (P < .05) as esterification of fat sources increased (HFA < HTHFA < HT). Acetate and A:P increased linearly (P < .10) but propionate and apparent total tract digestibility of OM decreased linearly (P < .05) as either saturation (T < PHT < HT) or esterification of fat sources increased. Ruminal NH3 N concentration increased linearly (P < .001) as saturation increased. Apparent ruminal digestibilities of ADF (P < .05) and NDF (P < .10) increased linearly as esterification increased. Flow of nonammonia nonmicrobial N to the duodenum was less (P < .10) but flow of microbial N was greater (P < .05) for the control diet than for fat-supplemented diets. Flows and small intestinal digestibilities of N and efficiencies of microbial protein synthesis were not altered by degree of saturation or esterification. Results confirm previous in vitro observations that T or HFA can alter ruminal digestion; however, because these effects usually are not observed in dairy cows, feed intake likely is very important in responses to supplemental fats. PMID- 9331888 TI - Effects of dietary nitrogen source and concentration in high-grain diets on finishing steer performance and nutrient digestion. AB - Two experiments were designed to evaluate dietary N source and concentration on finishing steer performance and nutrient digestion. In Exp. 1, 100 steers were used in a randomized complete block design experiment with 2 x 2 + 1 factorially arranged treatments. Diets contained 1.93 or 2.24% N supplemented by urea or soybean meal (SBM), or 2.24% N supplemented by cottonseed meal (CSM). Steers fed SBM-supplemented diets gained 13% faster (P < .01) and were 9% (P < .01) more efficient converting feed to gain than steers receiving urea. Steers fed diets containing 2.24% N were 4% (P < .05) more efficient than those fed diets containing 1.93% N. Steers fed CSM-supplemented diets gained 6% (P < .10) less efficiently than steers receiving SBM. Increasing dietary N with urea from 1.93 to 2.24% decreased carcass weights 3%, whereas increasing dietary N with SBM increased carcass weights 3%. Carcass-adjusted gains were reduced 8% by increasing urea from .9 to 1.5% but increased 7% by increasing SBM from 6.1 to 10.5% of DM. In Exp. 2, four ruminally and duodenally cannulated steers (390 kg) were used in a 4 x 4 Latin square design experiment to evaluate urea and SBM supplementation on digestion. Diets contained no supplemental N, 1.84% N with urea or SBM as the supplement, or 2.16% N with SBM as the supplement. Total tract starch digestion, duodenal microbial N flow, and efficiency of microbial protein synthesis in the rumen were higher (P < .10) in steers fed SBM- than in those fed urea-supplemented diets. Supplementation with SBM increased metabolizable protein supply and dietary energy utilization. PMID- 9331889 TI - Rapid communication: molecular cloning of the porcine beta 2-adrenergic receptor gene. PMID- 9331890 TI - Infection by Borrelia burgdorferi and cutaneous B-cell lymphoma. AB - In past years, association of primary cutaneous B-cell lymphoma (CBCL) with infection by Borrelia burgdorferi has been reported in a few patients. The evidence for a pathogenetic role was based on clinical grounds or raised titre of antibodies in serum. Both methods, however, do not prove the association between the micro-organism and the CBCL, especially in countries where infection by Borrelia burgdorferi is endemic. Moreover, the exact percentage of Borrelia burgdorferi-positive CBCL is not known. We retrieved from our files 50 cases of CBCL to perform PCR analysis of Borrelia burgdorferi DNA on paraffin-embedded tissue sections. Only patients with primary CBCL were selected. In all cases, monoclonality of the infiltrate was confirmed by immunohistological pattern of immunoglobulin light chains or molecular analysis of JH gene rearrangement, or both. Specific DNA sequences of Borrelia burgdorferi were identified in cutaneous lesions from 9 patients (follicle center lymphoma: 3/20; immunocytoma: 3/4; marginal zone B-cell lymphoma: 2/20; diffuse large B-cell lymphoma: 1/6). Specificity was confirmed by Southern blot hybridisation in all positive cases. We could show that Borrelia burgdorferi DNA is present in skin lesions from a small proportion of patients (18%) with various types of CBCL. Our results may have therapeutic implications. In analogy to Helicobacter pylori-associated MALT lymphomas, which in some cases can be cured by eradication of Helicobacter pylori infection, a proportion of CBCL may be cured with antibiotic therapy against Borrelia burgdorferi. Although yet speculative, adequate antibiotic treatment for patients with primary CBCL should be considered before more aggressive therapeutic options are applied, particularly in countries where infection by Borrelia burgdorferi is endemic. PCR analysis of Borrelia burgdorferi DNA is a fast test that should be performed in all patients with CBCL to identify those who more likely could benefit from an early antibiotic treatment. PMID- 9331891 TI - p53 and bcl-2 expression do not correlate with prognosis in primary cutaneous large T-cell lymphomas. AB - Overexpression of p53 protein in cutaneous T-cell lymphoma (CTCL) has been reported in primary cutaneous large T-cell lymphomas (PCLTCL) and has been associated with tumor progression and transformation in mycosis fungoides. However, the prognostic significance of p53 expression has not been studied thus far. In the present study we investigated the expression of p53 as well as bcl-2 protein in 27 PCLTCL, including 19 CD30-positive and 8 CD30-negative lymphomas, retrieved from the registry of the Dutch Cutaneous Lymphoma Working Group. The results were correlated with follow-up data and proliferative activity, as assessed by the percentage of MIB-1 positive tumor cells. Overexpression of p53 protein, defined as nuclear staining of more than 5% of the tumor cells, was found in 10 of 27 cases (37%), including 6 of 19 (32%) CD30+lymphomas and 4 of 8 (50%) CD30-PCLTCL. bcl-2 protein was expressed in 6 of 19 (32%) CD30+lymphomas and in only 1 of 8 (12%) CD30-PCLTCL. However, no significant correlation between p53 or bcl-2 expression and prognosis was found, neither in the whole group, nor within the CD30+ or CD30- group. In addition, no relationship between p53 expression and proliferative activity was found. The results confirm that p53 expression is more common in PCLTCL than in mycosis fungoides and Sezary syndrome. However, neither p53 nor bcl-2 expression correlated with survival or proliferative activity. PMID- 9331893 TI - Merkel cell hyperplasia in chronic radiation-damaged skin: its possible relationship to fibroepithelioma of Pinkus. AB - Moderate hyperplasia of Merkel cells (MC) in chronic sun-damaged skin and hypertrophic actinic keratoses is well known. In the present study we investigated the number of MC in 24 samples of chronic radiation dermatitis and 19 cases of fibroepithelioma of Pinkus (FP), which is known to arise preferably in radiation-damaged skin. Using antibodies against the low molecular weight cytokeratins 8, 18, and 20 and chromogranin A to visualize MC, we found hyperplasia of MC in chronic radiation dermatitis. Additionally, in all FPs we could detect many MC, especially in areas with a pronounced fenestrated pattern. Recently, regulative functions of MC on the growth of follicular epithelium under various conditions were discussed. Thus, MC hyperplasia suggests a causal role also in the development of FP. In this context, hyperplasia of MC in chronic radiation dermatitis could explain the frequent occurrence of FP due to radiation exposure. As we recently found MC also in trichoblastomas but not in basal-cell carcinomas, the MC in FP may indicate its relationship to the benign trichoblastoma rather than to the basal-cell carcinoma. It is possible that regulative influences of the MC are important for the clinically rather benign course of FP. PMID- 9331892 TI - CD56-positive (nasal-type T/NK cell) lymphoma arising on the skin. Report of two cases and review of the literature. AB - Several authors have reported cases of patients with malignant lymphoma with unique characteristics, designated nasal-type T/NK cell lymphoma, which expresses the natural killer (NK) cell marker and shows frequent extra-nodal involvement and poor prognosis. We report 2 cases of this type of lymphoma which were CD56 positive and showed a histopathologically angiocentric pattern with cutaneous and subcutaneous tumorous lesions. Patient 1 had extensive invasion of skin, underlying skeletal muscle, spleen and bone marrow, and died of sepsis 34 months after onset. Patient 2 had multiple subcutaneous nodules and invasion to mammary gland, lung, lymph node and spleen at the time of her first visit. She died of a rapid invasion of lymphoma cells to the liver 5 months after onset. Both patients showed similar immunophenotypes of tumor cells (CD2+, CD3-, CD4-, CD8-, CD20-, CD56+) and germ line configuration of the heavy chain of immunoglobulin (JH), T cell receptor C beta-1 subunit DNA and T-cell receptor J gamma subunit DNA. Epstein-Barr virus early regions RNA was demonstrated in the nuclei of tumor cells of both patients with in situ hybridization. The histopathological examination of the skin lesions of both patients revealed the features of angiocentric lymphoma. The detection of CD56 in the tumor cells of cutaneous lymphomas should be routinely performed for the early diagnosis of this type of lymphoma with extremely poor prognosis. PMID- 9331894 TI - Fibrohistiocytic differentiation in subcutaneous fatty tumors. Study of spindle cell, pleomorphic, myxoid, and atypical lipoma and dedifferentiated liposarcoma cases composed in part of CD34+ fibroblasts and FXIIIa+ histiocytes. AB - Subsets of dendritic cells, fibroblasts which express the human progenitor cell antigen CD34 or histiocytes which express coagulation factor XIIIa (FXIIIa), are present in fat and in collagenous connective tissue. As components of the microvascular unit, these fibrohistiocytic cell subsets may interact during stromal remodeling, repair, and neoplasia. We studied white fat and subcutaneous fatty tumors to determine if CD34 and/or FXIIIa+ "fibrohistiocytic" dendritic cell subsets are involved in their morphogenesis. Three lipomas (L), 1 intramuscular lipoma (IL), 1 myxoid lipoma (ML), 2 pleomorphic lipomas (PL), 2 spindle cell lipomas (SCL), 8 angiolipomas (AN) in 4 patients, 1 atypical lipoma/well-differentiated liposarcoma (AL), 1 de novo dedifferentiated liposarcoma (DL), and 1 recurring atypical myxoid signet ring lipomatous tumor were examined for CD34, FXIIIa and in some cases for CD31, desmin, Ki 67, or S 100. Normal fat has scattered CD34+ dendritic cells and small FXIIIa+ dendritic histiocytes among variably S-100+ adipocytes. The CD34 and FXIIIa+ dendritic cells are more numerous near vessels and within fibrovascular septae. In L and IL, CD34 and FXIIIa+ dendritic cells are activated and some adipocytes express CD34. Mesenchymal areas of SCL, PL, ML, and AL and DL are composed of CD34+ dendritic cells with CD34+ but FXIIIa-negative floret cells in PL or atypical cells in AL and DL. FXIIIa+ dendritic cells are numerous in these lesions, comprising 30-40% of cells in SCL and PL, and 50% in ML, AL, and DL. AN have focal CD34+ interstitial cells and plump FXIIIa+ cells that in one case resembled multivacuolated lipoblasts. The myxoid signet ring lipomatous tumor was CD34 negative with few FXIIIa+ cells. We conclude that subsets of CD34+ and FXIIIa+ dendritic microvascular cells are present in normal fat and proliferate together in various types of lipomas and in at least some dedifferentiated liposarcomas. PMID- 9331895 TI - Hypopigmented common blue nevus. AB - Blue nevus is a benign pigmented lesion of dermal melanocytes with a number of histologic and clinical variants, of which the major types are the common blue nevus, cellular blue nevus and combined nevus. This study describes 9 cases of hypopigmented blue nevus (HBN), a variant of common blue nevus in which there is minimal identifiable melanin pigment. We also discuss the usefulness of the immunoperoxidase stain HMB-45 in relation to the diagnosis of HBN and the lesions with which it may be histologically confused, namely common intradermal nevus, dermatofibroma, neurofibroma, dermal scar and desmoplastic malignant melanoma. The HMB-45 stain was found to be uniformly positive in all 9 cases of HBN, in contrast to the other dermal lesions which have been reported as either negative or showing only focal positivity. The physical distribution and age range of the patients in this study was similar to the age and sites for common BN, supporting the relationship between the 2 lesions. The occurrence of HBN in predominantly young adults indicates that this lesion is not a phenomenon due to ageing or degenerative change, and should be regarded as a variant of common blue nevus. PMID- 9331896 TI - Expression of integrin subunits and CD44 isoforms in psoriatic skin and effects of topical calcitriol application. AB - Increasing evidence suggests involvement of integrins and CD44 isoforms in the pathogenesis of psoriasis, contributing to uncontrolled keratinocyte proliferation, neovascularization, and invasion of inflammatory cells. We have analyzed immunohistochemically in situ expression of integrins (CD29, CDw49b, CDw49c, CDw49e, CDw49f) and CD44 isoforms (CD44 standard, CD44 var/v6, CD44 v10) on frozen sections of normal and psoriatic skin (nonlesional skin, lesional skin before and along with topical calcitriol treatment). We did not observe visual changes of immunoreactivity in normal as compared to nonlesional psoriatic skin, while the staining pattern of CDw49c, CDw49f, and CD29 was severely altered in untreated lesional psoriatic skin. Most markedly, CDw49c, CDw49f, and CD29 were focally upregulated in suprapapillar epidermal compartments of lesional psoriatic skin, a staining pattern that is in accordance with the phenomenon that was described by Pinkus as "squirting papilla". Additionally, an increased proportion of inflammatory and endothelial cells revealed immunoreactivity for CD44(std.) in untreated lesional psoriatic as compared to nonlesional psoriatic or normal skin. After 8 weeks of topical calcitriol treatment (15 micrograms/g ointment), the staining pattern for CDw49c, CDw49f and CD29 was markedly changed in epidermis of lesional psoriatic skin, reverting to the staining pattern characteristic for the nonlesional psoriatic or normal human skin, although epidermal expression of CDw49f was still upregulated and CDw49e-, CDw49f-, CD29-, and CD44(std.) immunoreactive inflammatory and endothelial cells were still to be found in the dermal compartment. PMID- 9331897 TI - Histopathological changes of primary HIV infection. Description of three cases and review of the literature. AB - The histopathological changes observed in the cutaneous rash of three patients who suffered the acute phase of HIV infection are described. In all three patients a perivascular and interstitial inflammatory infiltrate was present in the upper and mid-reticular dermis. In one biopsy isolated areas of epidermal necrosis were observed and in the two other biopsies a perifollicular inflammatory infiltrate was detected with perforation in one case. Furthermore, a periductal infiltrate was observed in one of these biopsies. PMID- 9331898 TI - The Muir-Torre syndrome in a black patient with AIDS: histopathology and molecular genetic studies. AB - In 1981, a black man had adenocarcinoma of the colon. In 1986, he had a sebaceous adenoma and the diagnosis of the Muir-Torre syndrome was established. The patient was found to be HIV sero-positive in 1986, and 8 years later fulfilled the CDC criteria for AIDS. During 1989 to 1993 the CD4 count was > 200 cells/ml and the patient had 2 sebaceous tumors, 1 basal cell carcinoma and 1 keratoacanthoma. In 1994 to 1996, the CD4 count was < 200 cells/ml and the patient developed 18 sebaceous tumors and a poorly differentiated adenocarcinoma of the finger which metastasized to axillary lymph nodes. Microsatellite analysis of tumor DNA from a sebaceous adenoma and adenocarcinoma of the finger revealed widespread microsatellite instability. The interaction of AIDS with the behavior of the tumors in the Muir-Torre syndrome has not previously been reported. Although our patient had an increase in the number of new sebaceous tumors at the same time he experienced deterioration of the immune system, he is doing well 15 years after resection of adenocarcinoma of the colon and 16 months after metastatic poorly differentiated adenocarcinoma of the skin. This follows the previously observed tendency for cancers of the Muir-Torre syndrome, especially those displaying widespread microsatellite instability, to be less lethal than their histologically similar counterparts in people without Muir-Torre syndrome. PMID- 9331899 TI - The use of nonneuronal cells for gene delivery. AB - The implantation of genetically engineered nonneuronal cells can provide an effective method for achieving localized delivery of discrete molecules to the CNS or for providing substrates for regrowth of neural structures. Most primary nonneuronal cells have the advantage of being easily obtainable from the prospective host for ex vivo retrovirus-mediated genetic manipulation (most will be mitotic in culture) and reimplantation as an autologous graft (circumventing the problem of immune rejection). As primary cells, they are unlikely to be tumorigenic. The most vexing problem for such systems remains the apparent loss of transgene expression from viral promoters after prolonged periods of engraftment. Much effort is currently being directed at optimizing sustained transgene expression by varying the promoters, by varying the cell types to be engineered, or by regulating expression by enhancing promoter function or substrate availability. While nonneuronal cells are excellent vehicles for achieving passive delivery of substances to the CNS, they lack the ability to incorporate into the host cytoarchitecture in a functional manner (e.g., make synaptic contacts). For this reason, not only may certain essential circuits not be re-formed, but the regulated release of certain substances through feedback loops may be missing. While apparently unimportant for some substances (e.g., ACh), for others (e.g., NGF), their unregulated, inappropriate, excessive, or ectopic release may actually be inimical to the host. Furthermore, the loss of foreign gene expression (the bane of gene therapy) may leave engineered nonneural cells incapacitated, whereas donor tissue originating from brain may intrinsically produce various CNS factors allowing correction to proceed despite inactivation of the introduced gene. In fact, CNS-derived tissue may provide as yet-unrecognized endogenous neuralspecific substances which are equally as beneficial to the host as the gene in question. Thus, future developments in gene delivery to the brain for some conditions may emphasize using neurons or neural progenitors for ex vivo genetic manipulation (Fisher, 1997) and refining techniques for the direct injection of therapeutic genes into neurons in vivo (see Snyder and Fisher, 1996). For a wide variety of conditions, however, using nonneuronal cellular vehicles or even nonbiologic synthetic vehicles may be efficient, effective, and safe strategies for the passive delivery of therapeutic molecules to discrete regions of the CNS. In fact, this approach may come closer than any other to immediate human applications. PMID- 9331900 TI - Frataxin shows developmentally regulated tissue-specific expression in the mouse embryo. AB - Friedreich ataxia (FRDA) is an autosomal recessive degenerative disease caused either by an intronic GAA triplet repeat expansion that suppresses the expression of the frataxin gene on chromosome 9q13, or, rarely, by point mutations in the frataxin gene. We investigated the expression of the mouse frataxin homologue during embryonic development by Northern blot analysis and RNA in situ hybridization. Very faint expression could be detected from E10.5 in the neuroepithelium and more clearly from E12.5 in the developing central nervous system. At E14.5, frataxin was expressed at a much higher level that remained constant into the postnatal period. Maximum expression was observed in the spinal cord, particularly at the thoracolumbar level, and in the dorsal root ganglia. Significant levels of transcript could also be detected in the proliferating cells in the periventricular zone, in the cortical plates, in the heart, in the axial skeleton, and in some epithelial and mesenchymal tissues. Overall, the distribution of frataxin mRNA was in good accordance with previous data from Northern analysis of adult human tissues, the major discrepancy being the expression in mouse embryonic cerebral cortex which was not observed in adult human brain. The tissues expressing frataxin during development appear to be those that become dysfunctional or atrophied in FRDA. Overall, our data suggest that frataxin is a protein whose expression is cell-specific and developmentally regulated. PMID- 9331901 TI - MPP+ induced substantia nigra degeneration is attenuated in nNOS knockout mice. AB - Recent studies showed that neuronal nitric oxide synthase (nNOS) plays a role in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine neurotoxicity. In the present study we examined the effects of striatal injection of 1-methyl-4-phenylpyridinium (MPP+) on substantia nigra degeneration in mutant mice lacking the nNOS gene or the endothelial nitric oxide synthase (eNOS) gene. Both striatal lesion volume and substantia nigra degeneration were significantly attenuated in the nNOS mutant mice but not in the eNOS mutant mice. The mice lacking nNOS showed a significant attenuation of MPP+(-) induced increases of 3-nitrotyrosine concentrations in the striatum. In a separate experiment administration of 7 nitroindazole for 48 h after MPP+ injections significantly attenuated substantia nigra degeneration in rats. Immunohistochemical studies showed apposition of nNOS positive neuronal processes on tyrosine hydroxylase-positive neurons. These results provide further evidence that neuronally derived NO and peroxynitrite play a role in MPP+ neurotoxicity. PMID- 9331902 TI - Survival-promoting activity of inhibitors of cyclin-dependent kinases on primary neurons correlates with inhibition of c-Jun kinase-1. AB - Cyclin-dependent kinases and mitogen-activated protein kinases have been implicated in the regulation of cellular survival and apoptosis. We tested the effect of two mitogen-activated/cyclin-dependent kinase inhibitors, olomoucine and butyrolactone I, on the in vitro survival of chick embryonic neurons. Sensory, sympathetic, and ciliary neurons, when prepared at their respective time point of programmed cell death, could be rescued from apoptosis by both inhibitors in a dose-dependent fashion. In contrast, dividing sympathetic precursors underwent apoptosis when treated with olomoucine, but not butyrolactone I, at the same range of concentration. With similar potency, olomoucine and butyrolactone I inhibited immunocomplex c-Jun kinase activity. Both substances inhibited neurite outgrowth in a dose-dependent manner; developmentally younger neurons were more sensitive to this effect than older ones. These results suggest that certain mitogen-activated/cyclin-dependent kinases associated with cell division in neuronal precursors (i) may become essential components of the apoptotic machinery by the time neurons reach their phase of naturally occurring cell death and (ii) may be necessary for neurite outgrowth during development. PMID- 9331903 TI - Neural transplantation: a hope for patients with Parkinson's disease. AB - More than 200 patients with Parkinson's disease (PD) have received intrastriatal grafts of human embryonic mesencephalic tissue. The clinical trials demonstrate that grafted dopamine (DA) neurons can survive in the human parkinsonian brain and reinnervate part of the host striatum. Long-term graft survival and function, at least up to 6 years after transplantation, is possible in PD despite a progressive degeneration of the patient's own DA neurons. A majority of patients with surviving grafts show long-term improvement of therapeutic value, but symptomatic relief is incomplete. Current research strategies to develop neural transplantation as a treatment for PD include (i) to increase DA neuron survival and density and extent of the dopaminergic reinnervation in the striatum; (ii) to implant DA neurons in denervated regions outside the caudate-putamen and to reconstruct the nigrostriatal pathway; and (iii) to find other sources of cells suitable for grafting. PMID- 9331904 TI - Cholecystokinin and enkephalin in brain stem pain modulating circuits. AB - Neurons in rostral ventromedial medulla and the periaqueductal gray modulate dorsal horn nociceptive transmission. Endogenous peptides implicated in this modulation include enkephalin (ENK), which is antinociceptive, and cholecystokinin (CCK), which has anti-opioid effects. In this study double-label fluorescence immunocytochemistry demonstrated somata and terminals with ENK- or CCK-like immunoreactivity in these regions. Although the distribution of CCK- and ENK-immunoreactive terminal fields overlapped significantly, co-localization was rare. Furthermore, CCK- and ENK-immunoreactive somata had different morphologies and distinct distributions. The overlap of CCK- and ENK- immunoreactive terminals arbors provides a morphological substrate for an antagonistic interaction of CCK and ENK within brainstem pain modulating circuits, as has been demonstrated in the spinal cord. PMID- 9331905 TI - Ultrastructural localization and regulation of protein gene product 9.5. AB - Protein gene product 9.5 (PGP), a ubiquitin hydrolase, is abundant in the nervous system. To investigate the ultrastructural localization of PGP and the regulation of its expression, we performed electron microscopic immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR) on normal and transected rat sciatic nerves. In normal nerves, strong PGP-immunoreactivity was localized in the myelinated and unmyelinated axons with virtually no staining in the Schwann cells. After nerve degeneration, denervated Schwann cells exhibited intense staining for PGP, corroborated with up-regulation of PGP transcripts by RT-PCR. The present data suggest that the pattern of expression of PGP is more complicated than was expected previously, and reflects the integrity of nerves and status of axon-Schwann cell interactions. PMID- 9331907 TI - Intra-hippocampal estradiol infusion enhances memory in ovariectomized rats. AB - Ovariectomized adult Long-Evans rats received an eight-trial training session in a hippocampal-dependent hidden platform water maze task. Following trial 8, rats received an intra-hippocampal injection of estradiol in a water soluble cyclodextrin inclusion complex (1.0, 2.0 or 5.0 micrograms/0.5 microliter), or saline. Twenty-four hours later, the retention test escape latencies of rats administered post-training intra-hippocampal injections of estradiol (5.0 micrograms) were significantly lower than those of saline treated rats, indicating a memory-enhancing effect of estradiol. Injections of estradiol (5.0 micrograms) given 2 h post-training had no effect on retention, indicating a time dependent effect of estradiol on memory storage processes. PMID- 9331906 TI - Expression of opioid-binding cell adhesion molecule (OBCAM) and neurotrimin (NTM) in E. coli and their reactivity with monoclonal anti-OBCAM antibody. AB - Opioid-binding cell adhesion molecule (OBCAM), neurotrimin (NTM) and limbic system-associated membrane protein (LAMP) are homologous and are the members of the IgLON family which is a subfamily within the immunoglobulin superfamily. We cloned the cDNAs for OBCAM and NTM, prepared recombinant proteins, and examined the reactivity of the previously prepared monocolonal anti-OBCAM antibody, OBC53, with the recombinant proteins by immunoblotting. These experiments revealed that OBC53 recognizes OBCAM about 1000 times as efficiently as NTM. Moreover, the NTM and LAMP peptides which have sequences homologous to the OBCAM peptide used for the preparation of OBC53 were 150 times less reactive to OBC53. Thus, the OBC53 antibody is a useful tool for specifically detecting OBCAM in immunochemical experiments. PMID- 9331908 TI - c-fos identifies GABA-synthesizing barosensitive neurons in caudal ventrolateral medulla. AB - Hypertension in the conscious rat, elicited by i.v. infusion of phenylephrine, evoked expression of the immediate early gene c-fos in discrete groups of brain stem neurons. Fos-immunoreactive neurons were located in the caudal ventrolateral medulla (CVLM); others were located in the nucleus of the tractus solitarius (NTS). Because of their sensitivity to alterations in arterial pressure, these neurons are likely to subserve the arterial baroreceptor reflex. The aim of this study was to identify the brain stem projections and the neurotransmitter content of the barosensitive CVLM neurons using neuronal tracing and immunohistochemistry. Some of the barosensitive CVLM neurons projected directly to the rostral ventrolateral medulla (RVLM), and many contained the GABA synthesizing enzyme, glutamic acid decarboxylase (GAD). Other CVLM neurons, containing markers of glutamate or catecholamine synthesis, were insensitive to baroreceptor stimulation. This study delineates neuronal pathways acting in the arterial baroreceptor reflex and identifies precisely GABA-synthesizing CVLM neurons as the source of inhibitory input to the RVLM. PMID- 9331909 TI - Five inhibitory transmitters coexist in pelvic autonomic vasodilator neurons. AB - Here we describe the localization of a potent vasodilator, calcitonin gene related peptide (CGRP), in pelvic autonomic neurons containing four other inhibitory transmitters: vasoactive intestinal peptide (VIP), neuropeptide Y, nitric oxide and acetylcholine. These neurons mediate endothelium-independent vasodilation by releasing nitric oxide and one or more neuropeptides. Sixty percent of nerve cell bodies in guinea-pig paracervical ganglia with immunoreactivity (IR) for VIP, choline acetyltransferase (ChAT) and nitric oxide synthase (NOS), also contained IR for CGRP. Furthermore, many VIP-IR varicose nerve terminals at the adventitia-medial junction of the guinea-pig uterine artery contained IR for CGRP, ChAT and NOS. Both alpha-hCGRP and beta-hCGRP were potent dilators of the uterine artery (pD2 values 8.1, 8.3, respectively), but 1 microM hCGRP(8-37) did not antagonize dilations produced by either agonist. Dilations produced by alpha-hCGRP were unaffected by removal of the endothelium. Taken together with results of our previous studies, we propose that CGRP can contribute directly to autonomic vasodilation, possibly via CGRP2 receptors on smooth muscle cells, and that CGRP is the fifth inhibitory transmitter co existing in pelvic vasodilator neurons. PMID- 9331910 TI - Combined event-related fMRI and EEG evidence for temporal-parietal cortex activation during target detection. AB - Target detection is the process of bringing a salient stimulus into conscious awareness. Target detection evokes a prominent event-related potential (ERP) component (P3) in the electroencephalogram (EEG). We combined the high spatial resolution of functional magnetic resonance imaging (fMRI) with the high temporal resolution of EEG to investigate the neural generators of the P3. Event-related brain activation (ERBA) and ERPs were computed by time-locked averaging of fMRI and EEG, respectively, recorded using the same paradigm in the same subjects. Target detection elicited significantly greater ERBAs bilaterally in the temporal parietal cortex, thalamus and anterior cingulate. Spatio-temporal modelling of ERPs based on dipole locations derived from the ERBAs indicated that bilateral sources in the temporal-parietal cortex are the main generators of the P3. The findings provide convergent fMRI and EEG evidence for significant activation of the temporal-parietal cortex 285-610 ms after stimulus onset during target detection. The methods developed here provide a novel multimodal neuroimaging technique to investigate the spatio-temporal aspects of processes underlying brain function. PMID- 9331912 TI - Simultaneous temporal processing is sensitive to prenatal choline availability in mature and aged rats. AB - Rats were trained at 2-4 months and at 24-26 months of age on a peak-interval timing procedure in which auditory and visual stimuli signaled two different fixed-interval schedules of reinforcement (15 and 30 s) that were presented simultaneously in a hierarchical fashion. Compared with control rats, increases in the probability of attention to the 15 s signal were observed for both the choline-supplemented and the choline-deficient rats. In contrast, an increase in attention to the 30 s signal was only observed for the choline-supplemented rats, whereas choline-deficient rats exhibited a decrease in attention that increased with age. Proportional rightward shifts in the remembered times of reinforcement emerged for the 24-26-month-old rats in the choline-deficient and control groups, but not in the choline-supplemented group. These results indicate that prenatal choline supplementation facilitates cognitive function across the lifespan, whereas prenatal choline deficiency impairs divided attention and accelerates age related declines in temporal processing. PMID- 9331911 TI - Botulinum toxin in upper limb spasticity: study of reciprocal inhibition between forearm muscles. AB - To establish whether botulinum A toxin (BTX-A) acts on modifying reciprocal inhibition between forearm muscles in spasticity, 20 patients with post-stroke upper limb spasticity lasting for more than 1 year were studied. Clinical examination, physiotherapeutic evaluation, standardized video-tape assessment and electrophysiological testing (flexor carpi radialis muscle M and H responses with study of reciprocal inhibition) were performed at baseline and 2 weeks, 1, 2, 3, 4 months after BTX-A treatment. BTX-A induced a significant decrease of tone and an improvement of motility and functional status, with a significant decrease of the M wave and the H reflex. The reduction in both inhibitory phases of reciprocal inhibition did not change after BTX-A treatment differently from that reported in upper limb dystonia. These findings indicate that the efficacy of BTX A in upper limb spasticity is mainly due to peripheral effects. PMID- 9331913 TI - Perinatal choline supplementation increases the threshold for chunking in spatial memory. AB - Chunking and perinatal choline supplementation each provide rats with alternative memory processing advantages. Evidence from radial-arm maze performance of adult (2- to 5-month-old) rats indicates that chunking of multiple food types (sunflower seeds, Noyes pellets and rice puffs) emerges for stable, differentiable baiting patterns as a function of the memory load (6, 12, 18 or 24 maze arms). The number of maze arms appeared to determine both the level of task difficulty at which rats began to implement a chunking strategy as well as when they were unable to successfully implement such a strategy due to the excess memorial demands of the task. In comparison to control rats, rats treated perinatally with choline supplementation displayed a horizontal rightward shift of the response function that related level of clustering of like-food types to the number of maze arms. These results indicate a higher threshold for implementing a chunking strategy in rats treated perinatally with choline supplementation, possibly due to a choline-induced increase in memory capacity. PMID- 9331914 TI - Reduction of NMDA-induced Ca2+ transients by a mu-opioid receptor agonist in dorsal horn neurons. AB - The effect of DAMGO, a mu-opioid receptor agonist, on intracellular Ca2+ transients evoked by the application of NMDA, was studied in freshly dissociated neurons from the superficial dorsal horn in the spinal cord of young rats. DAMGO (5-10 microns) reduced the amplitude of the Ca2+ transients measured with Fura-2 to 58 +/- 17% of the controls in 41% of the neurons tested. The effect of DAMGO was dose dependent and reversible. The reduction of NMDA-induced Ca2+ transients by DAMGO was prevented by application of the opioid antagonists naloxone (0.1-5 microM) and CTAP (0.2-2 microM). DAMGO also reduced Ca2+ transients induced by high K+ in 29% of the neurons. These data suggest that mu-receptor activation regulates NMDA-induced Ca2+ transients in a complex manner, by reducing both a depolarization-induced component and the NMDA-channel component of this Ca2+ signal. PMID- 9331915 TI - Modification of glutamine synthetase expression by mammalian Muller (glial) cells in retinal organ cultures. AB - One of the key enzymes in glial-neuronal transmitter recycling is glutamine synthetase (GS). In the retina, GS is exclusively expressed by glial (Muller) cells where it serves to convert neuron-released active transmitter substances (glutamate and GABA) into glutamine. Experiments on avian retinae have shown that GS expression is developmentally regulated by glucocorticoid hormones and, to a lesser extent, by a non-hormonal control mechanism(s). Much less is known about GS regulation in mammalian retinae, although either increases or decreases of GS immunoreactivity have been observed in Muller cells in different forms of retinal pathologies. We studied GS expression in postnatal rabbit retinae both in vivo and explanted as wholemounts in vitro, using immunocytochemistry and Western immunoblotting. GS expression was detectable in vivo from the fourth postnatal day, and increased rapidly within the first weeks of life. Levels were lower in vitro than in vivo by an order of magnitude, and could be significantly stimulated (> 60-110%) in vitro by application of hydrocortisone, conditioned medium from cultured retinal pigment epithelium and glutamate or ammonia, but not GABA. It is concluded that GS expression in mammalian Muller cells is dependent on systemic control by glucocorticoid hormones, as observed in birds, but environmental (activity-dependent) factors may play a more important role in mammals. PMID- 9331916 TI - Olfactory cues from an oxytocin-injected male rat can induce anti-nociception in its cagemates. AB - We recently demonstrated an olfactorily induced tail skin temperature drop in saline-injected rats exposed to an oxytocin-injected cagemate, an effect abolished by olfactory impairment. Treatment with oxytocin may induce both nociceptive and anti-nociceptive effects. The contrasting effects likely depend on the model and dosage used. Here we report an increased hindpaw withdrawal latency in response to nociceptive heat following the subcutaneous administration of oxytocin (1 mg/kg). An increased withdrawal response latency was also found in the untreated cagemates of an oxytocin-treated rat. The anti-nociceptive effect was abolished in oxytocin-antagonist-injected cagemates. Our results suggests that an olfactorily induced oxytocinergic mechanism is activated in the cagemates of an oxytocin-injected rat promoting anti-nociception. PMID- 9331917 TI - Induction of the dual specificity phosphatase PAC1 in rat brain following seizure activity. AB - Recurrent seizure activity leads to delayed neuronal death as well as to inflammatory responses involving microglia in hippocampal subfields CA1, CA3 and CA4. Since mitogen activated protein (MAP) kinases control neuronal apoptosis and trigger generation of inflammatory cytokines, their activation state could determine seizure-related brain damage. PAC1 is a dual specificity protein phosphatase inactivating MAP kinases which we have found to be undetectable in normal brain. Despite this, kainic acid-induced seizure activity lead to rapid (approximately 3 h) but transient appearance of PAC1 mRNA in granule cells of the dentate gyrus as well as in pyramidal CA1 neurons. This pattern changed with time and after 2-3 days PAC1 was induced in dying CA1 and CA3 neurons. At this time PAC1 mRNA was also expressed in white matter microglia as well as in microglia invading the damaged hippocampus. PAC1 may play an important role controlling MAP kinase involvement in both neuronal death and neuro-inflammation following excitotoxic damage. PMID- 9331918 TI - Modulation of septal cell activity by extracellular zinc. AB - Zinc released from axon terminals in the brain can interact with multiple membrane channels and receptors. However, the specific effects of these Zn(2+) dependent interactions on physiological processes remains unclear. Because Zn(2+) containing axon terminals are abundant in the septal region, we selected a septal cell line (SN56) to study the effects of Zn2+ on cell activity. Voltage-clamp recordings showed well-developed voltage-dependent Na+, Ca2+ and K+ currents. Micromolar concentrations of Zn2+ partially blocked Na+ and Ca2+ currents without affecting K+ currents. Current-clamp recordings showed that SN56 cells fire spontaneous and evoked action potentials. While most (> or = 83%) Na+ and Ca2+ currents were blocked with 1 microM tetrodotoxin (TTX) and 2 mM Co2+, action potentials persisted after either 1 microM TTX or 2 mM Co2+ application. In contrast, concentrations of Zn2+ (50-300 microM) that induced incomplete blockade (< or = 50%) of either Ca2+ and Na+ currents abolished action potential generation. These data show that simultaneous and partial blockade of Ca2+ and Na+ channels by Zn2+ inhibit SN56 cell activity. Because septal outputs extensively modulate the excitability of cortical and subcortical brain regions, Zn2+ inhibition of action potential generation in septal neurons could play an important physiological role in regulating brain activity. PMID- 9331919 TI - Propofol produces anticonflict action by inhibiting 5-HT release in rat dorsal hippocampus. AB - We examined the effect of propofol, an injectable anesthetic agent on conflict behavior in a Vogel type conflict test and on release of serotonin (5 hydroxytryptamine, 5-HT) in the dorsal hippocampus using an in vivo microdialysis method in rats. Propofol (20 and 40 mg/kg, i.p.) dose-dependently suppressed elevated 5-HT release normally seen in a conflict situation and concomitantly attenuated conflict behavior. These findings suggest that propofol exerts an antianxiety action by inhibiting 5-HT neuronal activity in the dorsal hippocampus. PMID- 9331920 TI - Brain activity assessed with PET during recall of word lists and narratives. AB - This study investigated the functional neuroanatomy involved in retrieval of structured versus unstructured verbal information. We compared cerebral blood flow using PET with the [15O]water method while subjects engaged in recall of novel and practised narratives and lists of unrelated words. Left orbital frontal cortex was activated during recall of both novel and practised unrelated words. Right parietal cortex was relatively more active during recall of the novel word list. Right orbital frontal cortex and anterior cingulate were relatively more active during recall of the practised but not the novel word list. These results are consistent with the role of left orbital frontal cortex in retrieval of unstructured verbal information. Right orbital frontal activity suggests that cognitive strategies may be involved in retrieval of well-practised words. PMID- 9331921 TI - Expression of glial cell line-derived neurotrophic factor (GDNF) mRNA following mechanical injury to mouse striatum. AB - Although glial cell line-derived neurotrophic factor (GDNF) expression is low in the adult brain, its administration protects dopaminergic neurons against a range of insults, leading to the suggestion of a role in dopaminergic regeneration. If locally produced GDNF is to fulfil a role in dopaminergic regeneration after injury, it seems reasonable to hypothesize that its expression will increase after mechanical trauma. We have demonstrated that GDNF mRNA expression increases within 6 h of using a wire knife to injure adult mouse striatum. Expression doubles after 1 week and remains elevated for at least 1 month. Most GDNF expression is associated with haemosiderin-containing cells, indicating production by brain macrophages. GDNF production by macrophages may be essential for neural regeneration following CNS trauma. PMID- 9331923 TI - Glycine receptor regulation of neurokinin1 receptor function in rat dorsal horn neurones. AB - Activation of spinal neurokinin1 (NK1) receptors leads to increases in the extracellular concentration of glycine in the dorsal horn. We have investigated the role of the inhibitory glycine receptor as a regulator of NK1 receptor mediated effects on dorsal horn neurones. Ionophoretic application of GR82334, a selective NK1 antagonist, did not alter dorsal horn neuronal activity evoked by cutaneous applications of mustard oil. However, in the presence of the glycine antagonists, strychnine or phenylbenzene-omega-phosphono-alpha-amino acid (PMBA), GR82334 displayed inhibitory properties. Therefore inhibitory glycine receptors may mask the contribution made by NK1 receptors to nociceptive processing. This is discussed with reference to the role of NK1 receptors during brief and long duration nociceptive transmission. PMID- 9331922 TI - Plasticity of motor organization in children and adults. AB - We explored the effects of maturational plasticity on motor activations for the affected hand in patients with unilateral lesion involving the rolandic cortex. Ten patients with early lesion (onset < 4 years), seven patients with late lesion (onset > or = 10 years) and eight normal adults underwent [15O]-water positron emission tomography (PET). Rolandic activations in the contralesional hemisphere were enhanced in both patient groups when compared to normal adults. Secondary motor and frontoparietal nonmotor cortices were more activated in the early than in the late lesion group, suggesting a greater potential for reorganization during early development than later in life. Cerebellar activations were similar in late lesion patients and normal adults, but significantly weaker in early lesion patients. PMID- 9331924 TI - The effects of subthreshold visual stimulation on P300 response. AB - The effects of the subthreshold visual stimuli on the electrical activity of the brain were investigated by an oddball-like paradigm and a single-stimulus paradigm. In the oddball-like paradigm, suprathreshold and subthreshold stimuli were presented randomly where the suprathreshold stimulus probability was 0.24. The single-stimulus paradigm contained only the suprathreshold stimuli and was identical to the oddball-like paradigm apart from the absence of subthreshold stimuli. Suprathreshold P3b amplitude was larger for the oddball-like than for the single-stimulus condition particularly in the centro-parietal region. This result suggests that at least one component of the P300 wave could be assigned to an automatic comparison and memory updating process which runs unconsciously. PMID- 9331925 TI - Changes in the alpha 2-adrenergic receptor subtypes gene expression in rat dorsal root ganglion in an experimental model of neuropathic pain. AB - We examined changes in expression of genes coding for alpha 2-AR subtypes in the dorsal root ganglion (DRG) in a rat model (spinal nerve ligation) or neuropathic pain. The present study demonstrates that the majority of DRG neurons express alpha 2C-AR mRNA and a small proportion of neurons express alpha 2A-AR mRNA, while few neurons express alpha 2B-AR mRNA in non-operated animals. In addition, alpha 2C- and alpha 2A mRNA levels in the DRG showed a significant decrease and increase, respectively in ligated animals. These findings suggest that alpha 2A- and alpha 2C-ARs in the DRG may play an important role in generating sympathetically maintained neuropathic pain. PMID- 9331926 TI - Expression of leptin receptor mRNA (long form splice variant) in the human cerebellum. AB - Primers specific to the long isoform of leptin receptor (OB-Rb) mRNA were used in reverse transcriptase-linked polymerase chain reactions (RT-PCR) to investigate the expression of this receptor in the hypothalamus and cerebellum of human and rat. For both regions, we observed RT-PCR cDNAs as well as restriction enzyme cleavage fragments of expected sizes. Additionally, in situ hybridization of human cerebellum using two independent [35S]oligonucleotide probes complementary to the OB-Rb mRNA sequence revealed a prominent hybridization signal within the granular layer. Overall, our findings demonstrate the expression of OB-Rb mRNA in the cerebellum and suggest that in such a location, leptin receptors may mediate a function presumably not linked to body weight homeostasis. PMID- 9331927 TI - Expression of corticotropin-releasing factor in the peripheral nervous system of the rat. AB - The occurrence and distribution of corticotropin-releasing factor (CRF) in the rat peripheral nervous system was studied by immunohistochemistry. CRF-positive nerve fibers were identified in the spleen, thymus, synovial membrane of the knee joint and adrenal gland. In general, CRF-positive fibers were seen predominantly in and around the blood vessels; however, many non-vascular thin varicose fibers were also observed. The neuronal character of the immunoreactive fibers was confirmed by staining consecutive tissue sections with a general neuronal marker, protein gene product 9.5. The finding of CRF-positive nerve fibers in the periphery demonstrates a strong anatomical link between the nervous, endocrine and immune systems, and may have pathophysiological implications in the inflammatory and stress-related disorders. PMID- 9331928 TI - Spinal analgesic actions of the new endogenous opioid peptides endomorphin-1 and 2. AB - Two highly-selective mu-opioid receptor agonists, endomorphin-1 and -2, were recently purified from bovine brain and are postulated to be endogenous mu-opioid receptor ligands. We sought to determine the effects of these ligands at the spinal level in mice. Endomorphin-1 and -2 produced short acting, naloxone sensitive antinociception in the tail flick test and inhibited the behavior elicited by intrathecally injected substance P. Both endomorphin-1 and -2 were anti-allodynic in the dynorphin-induced allodynia model. Although acute tolerance against both endomorphins developed rapidly, endomorphin-1 required a longer pretreatment time before tolerance was observed. We conclude that the endomorphins are potent spinal antinociceptive and anti-allodynic agents and that they or related compounds may prove therapeutically useful as spinal analgesics. PMID- 9331929 TI - Hemispheric asymmetry in the visual contribution to postural control in healthy adults. AB - This study was carried out in order to test the hypothesis of a right hemisphere dominance in the visual control of body balance. Eight healthy adults were subjected to a self-regulated lateral balance task, performed while sitting on a rocking platform. Four visual conditions were tested: open eyes with normal vision, closed eyes in the dark, left visual field-right hemisphere and right visual field-left hemisphere. Head and support displacements in the roll plane were recorded by means of an optoelectronic system. Two main results emerged from this study: (1) head stabilization in space was much more efficient in the left visual field-right hemisphere condition than in the three other visual conditions, and (2) although vision played an important role in the body stability whatever the anatomical level, there was no right hemisphere dominance at the pelvic level. A clear right hemisphere dominance was thus demonstrated as regards the visual contribution to head stabilization in space. PMID- 9331930 TI - Amygdala beta-noradrenergic influence on hippocampal long-term potentiation in vivo. AB - We have previously shown that hippocampal long-term potentiation (LTP), one form of synaptic plasticity that may underlie learning and memory, is attenuated by blocking neuron activity of the basolateral amygdala (BLA). In the present study we investigated the amygdala noradrenergic or cholinergic contribution to hippocampal LTP formation. When propranolol, a beta-adrenoceptor antagonist, was injected into the BLA 10 min before tetanus, the formation of LTP in the perforant path-dentate granule cell synapses was significantly impaired. Scopolamine, a muscarinic cholinergic receptor antagonist, did not affect the formation of LTP. These results suggest that amygdala beta-noradrenergic activity plays a critical role in modulation of hippocampal LTP. PMID- 9331931 TI - A1 noradrenergic modulation of AV3V inputs to PVN neurosecretory cells. AB - Phasically active neurosecretory neurons in the paraventricular nucleus (PVN) of urethane-anesthetized rats displayed orthodromic excitation, inhibition or no response following electrical stimulation of the anteroventral third ventricle (AV3V) region, and exhibited orthodromic excitation or no response following electrical stimulation of the A1 noradrenergic region of the ventrolateral medulla. Of the 14 neurons that responded to both the stimuli, A1 region stimulation at the subthreshold current significantly enhanced the excitation induced by AV3V region stimulation, and the enhancement was abolished by iontophoretically applied phentolamine, an alpha-adrenoceptor antagonist, but not by timolol, a beta-adrenoceptor antagonist. These results suggest that A1 noradrenergic projections may act to potentiate the excitatory inputs from the AV3V region to vasopressinergic PVN neurons through alpha-adrenoceptor mechanisms. PMID- 9331932 TI - Use of paper for treatment of a peripheral nerve trauma in the rat. AB - Reinnervation of the muscles and skin in the rat hindpaw was studied after transection and attempted repair of the sciatic nerve. Reconnecting the transected nerve with lens cleaning paper was at least as effective in rejoining the transected nerves as traditional microsurgical neurorraphy. Paper induced a slightly bigger fibrous scar around the site of transection than neurorraphy, but this scar did not cause impairment of functional recovery or excessive signs of neuropathic pain. We conclude that a paper graft can be used in restorative surgery of severed peripheral nerves. PMID- 9331933 TI - Enhancement of neurite outgrowth by the soluble form of human L1 (neural cell adhesion molecule). AB - L1, a neural cell adhesion molecule, is involved in neurite outgrowth, migration and fasciculation. Although L1 is a membrane glycoprotein expressed on neural cells, the soluble form of L1 is generated in vivo by proteolysis. In the present study, a stable transfectant of Chinese hamster ovary (CHO) cells secreting human L1 without cytoplasmic and membrane spanning domains was generated, and the function of the secreted L1 was examined. Explants from embryonic chick brain stem were cultured on a substrate coated with polyethylenimine (PEI) alone, on substrate-bound L1 or in medium containing soluble L1. The neurites induced by L1, both the substrate-bound form and the soluble form, were 2-3 times longer than those cultured on PEI. The ability of the soluble L1 to induce neurite formation was slightly greater than that of the substrate L1. The present results demonstrated that neurite outgrowth was induced not only by substrate-bound L1 but also by soluble L1. Soluble L1 could be a pharmaceutical candidate for the promotion of nerve regeneration. PMID- 9331934 TI - IL-1-induced iNOS expression in human astrocytes via NF-kappa B. AB - Nitric oxide (NO) plays an important role in host defense as well as cell injury within the CNS. In contrast to rodent species, human astrocytes are the major glial source of NO. Although interleukin (IL)-1 stimulates astrocyte inducible NO synthase (iNOS) expression, the mechanism is poorly defined. In the present study using primary human fetal astrocyte cultures, we found that IL-1 beta stimulated activation of nuclear factor kappa B (NF-kappa B) within 2 h, iNOS mRNA expression at 8 h, and maximal NO production by 5 days post-treatment. This IL-1 induced activation of astrocyte iNOS was suppressed by pyrrolidine dithiocarbamate, an inhibitor of NF-kappa B activation, suggesting involvement of a NF-kappa B mechanism. PMID- 9331936 TI - Orientation-dependent binocular interactions in area 21a of the cat. AB - We investigated binocular interactions in area 21a cells of the anaesthetized cat. Visual stimuli were drifting sinusoidal gratings presented at the same optimal orientation in each eye (iso-oriented condition) or at the optimal orientation in the dominant eye and the orthogonal orientation in the other eye (orthogonal condition). In 68% of cells the response in the binocular iso oriented condition was greater than the dominant eye monocular response, while in 88% of cells the response in the binocular orthogonal condition was less than the dominant eye monocular response. Our results suggest a possible role for this extrastriate region of cortex in binocular contour rivalry. PMID- 9331935 TI - Presence of mu and kappa opioid receptor mRNAs in galanin but not in GnRH neurons in the female rat. AB - Using a combination of radioactive and non-radioactive in situ hybridizations, the expression of mu and kappa opioid receptor mRNA was investigated in neurons in the female rat preoptic nucleus expressing galanin and gonadotropin-releasing hormone (GnRH) mRNA. Numerous cells expressing both mu or kappa and galanin were found in the intermediate and rostral regions of the preoptic area whereas little co-localization was observed at the rostral level. The number of kappa/galanin expressing cells was greater than that of mu/galanin cells. mu/galanin co localization was observed essentially in the anteroventral preoptic nucleus while neurons expressing kappa/galanin were present in both the anteroventral preoptic nucleus and in the periventricular hypothalamic nucleus. Co-localization of GnRH with mu or kappa could not be detected in the preoptic area. These observations showed that galaninergic neurons but not GnRH neurons of the preoptic area might be directly regulated by endogenous opioid peptides. PMID- 9331937 TI - Auditory response areas altered by intermodulation distortion products from background tones. AB - Response areas (RAs) of sensory neurones are dynamically modified by attention, denervation of specific afferent input, blocking inhibition, and by prolonged conditioning with extra-RA stimuli. Here we demonstrate in auditory neurones that the RA is also critically influenced by the background to stimuli. When RAs are measured in the presence of non-excitatory extra-RA tones, new RAs arise at frequencies otherwise not excitatory, as a consequence of non-linear receptor organ transduction. The new RAs can become more sensitive than the RA in quiet conditions such that neurones are then effectively tuned to a new frequency. Thus, even in a modestly complex environment, auditory neurones do not signal a fixed range of sounds but effectively code sounds to which they are otherwise unresponsive. PMID- 9331944 TI - Heirs of the jaguar and the anaconda: HLA, conquest and disease in the indigenous populations of the Americas. PMID- 9331945 TI - Episodic evolution and turnover of HLA-B in the indigenous human populations of the Americas. AB - Nucleotide sequences were determined for the HLA-A, B and C alleles of three populations of Amerindians: the Havasupai tribe from North America, and the Guarani and Kaingang tribes from South America. All 15 Havasupai alleles are found in Eastern Hemisphere populations, whereas the Guarani and Kaingang each have six alleles that appear to be present only in the Western Hemisphere. Nine of the "new" alleles come from HLA-B, one comes from HLA-A and one from HLA-C: ten appear to be the result of recombination and one the result of point substitution. Of the 14 Guarani alleles and 16 Kaingang alleles, only four are held in common. Despite their differences, the three tribes possess comparable numbers of HLA class I alleles, revealing a trend for "allele turnover", in which new alleles tends to supplant older alleles rather than supplement them. Although many new HLA-B alleles have been produced in Latin America, their net effect has been to differentiate populations, not to increase allele diversity within a population. From sequence comparisons, the Amerindian subset of HLA class I allotypes appears to cover the overall ranges of peptide binding specificity, natural killer-cell interactions, and CD8 interactions, that are found in all HLA class I. The recombinations that produced the new alleles of the Kaingang and Guarani class I are predicted to have modulated these functional properties rather than radically change them. Exchange of Bw4 and Bw6 motifs by recombination are noticeably absent in the events forming new alleles in America, whereas they have been the most common of recombinations elsewhere. PMID- 9331946 TI - Dissimilar evolution of B-locus versus A-locus and class II loci of the HLA region in South American Indian tribes. AB - Native American populations have a limited HLA polymorphism compared with other ethnic groups. In spite of this, many novel HLA-B locus alleles, not observed in other populations, have been identified in South American tribes, and rapid evolution of this locus has been suggested. We have studied unrelated subjects of the Toba (TOB n = 116), Wichi (WIC n = 46) and Pilaga (PIL n = 14) tribes from northeastern Argentina to investigate the extent of the HLA polymorphism and obtain clues of selective forces that may have acted in these populations. In these tribes the number of HLA alleles is small at all loci except HLA-B, which presents 22 alleles. Seven novel alleles were characterized including 5 of HLA-B (B*35092, B*3518, B*3519, B*4009, B*4803) 1 at HLA-A (A*0219) and 1 at DRB1 (DRB1*0417). All these variants may have arisen by gene conversion events. Some of the novel variants represent the most frequent alleles of these populations (B*4803 in TOB and PIL; B*3519 in WIC) or are the most frequent subtypes in their lineages. HLA-A, B, DRB1,DQA1 and DQB1, but not DPB1, display relatively similar gene frequencies. This results in high heterozygosity in all the tribes for all the loci studied except HLA-DPB1. The larger polymorphism and the generation and maintenance of novel alleles at the HLA-B locus suggests a more specialized response of this locus to evolutionary forces. These effects may be related to the nature of the polymorphism, to the number of founder alleles and to the functional characteristics of the individual alleles. PMID- 9331947 TI - Further diversification of the HLA-B locus in Central American Amerindians: new B*39 and B*51 alleles in the Kuna of Panama. AB - Several new HLA-B locus alleles have been discovered in South American Amerindians. By contrast, analysis of the MHC class I alleles of North American native populations has revealed few new HLA-B alleles. This suggests that the HLA B locus is evolving rapidly in South American populations. Here we describe the HLA-B locus alleles present in individuals from a Central American tribe, the Kuna of Panama. Using a sequence-based typing technique that separates alleles by denaturing gradient gel electrophoresis (DGGE) followed by direct sequencing, we determined the HLA-B alleles from eight Kunas. Two of the HLA-B alleles present in the Kuna have been previously described in other South American Amerindian populations; one allele has been characterized in a Mexican-American. We characterized two new HLA-B alleles in the Kuna, HLA-B*3911 and HLA-B*5110. HLA B*3911 differed from HLA-B*3905 by only a single nucleotide substitution in exon 3. This substitution resulted in an amino acid replacement of leucine by arginine at residue 156 in the alpha 2 domain. Such a change may affect the repertoire of peptides that are bound by this molecule. HLA-B*5110 differed significantly from other HLA-B*51 alleles in that it is the result of an unusually large intra-locus recombination event of minimally 216 nucleotides. This recombination results in an allele that is part HLA-B*51 and part HLA-B*40. Thus, more dramatic recombination events may also play a role in the rapid evolution of the HLA-B locus in Amerindians. PMID- 9331948 TI - Interactions of HLA-B*4801 with peptide and CD8. AB - Functional properties of the B*4801 allotype were investigated using HLA class I deficient 221 cells transfected with B*4801 cDNA. From pool sequence analysis of endogenously bound peptides, B*4801 was shown to select for nonamer peptides having glutamine or lysine at position 2 and leucine at the carboxyl-terminus. In an in vitro cell-cell binding assay, B*4801 binds CD8 alpha homodimers weakly due to the presence of a threonine residue at position 245 in the alpha 3 domain. A mutant B*4801 molecule in which alanine replaces threonine 245, binds CD8 alpha homodimers at levels comparable to those of other HLA class I allotypes. Despite the low affinity of B*4801 for CD8 alpha, alloreactive T-cells that recognize B*4801 molecules expressed by the 221 transfectant are inhibited by anti-CD8 monoclonal antibodies. Analysis of 25 B*48-expressing individuals from various populations showed threonine 245 was encoded by every B*48 allele. PMID- 9331950 TI - DNA typing of HLA class I (HLA-A) and class II genes (HLA-DR, -DQ and -DP) in Japanese patients with gastric cancer. AB - Gastric cancer is multifactorial disease and several reports have described genetic factors involved in pathogenesis of gastric cancer. Recently, it was reported that HLA class II gene DQB1*0301 was strongly associated with gastric cancer in Caucasian population (20). We performed DNA typing of HLA class I (HLA A) and HLA class II genes (HLA-DR, DQ and DP) to elucidate the HLA alleles or HLA haplotypes associated with gastric cancer in Japanese population using polymerase chain reaction-sequence-specific oligonucleotide probe analysis in 88 unrelated patients with gastric cancer and 525 unrelated healthy controls. We observed slight difference in frequencies of some HLA alleles and haplotypes between gastric cancer patients and controls; however, after Bonferroni correction, statistical significance was not confirmed. It is possible that environmental factors such as diet cover the contribution of genetic factors to the disease in Japanese population, which has a higher frequency of gastric cancer than do Caucasian populations, most likely due to more exposure to environmental risk factors. PMID- 9331949 TI - Mapping epitopes to distinct regions of the extracellular domain of endoglin using bacterially expressed recombinant fragments. AB - Endoglin (CD105) is a homodimeric cell surface component of the TGF-beta 1 receptor complex, which is expressed at high levels on vascular endothelium and at lower levels on activated monocytes. It is also the target gene for the dominantly inherited vascular disorder hereditary hemorrhagic telangiectasia type 1. To date, each family has a distinct endoglin mutation, most of which generate premature stop codons. The purpose of the current study was to identify monoclonal antibodies capable of binding to normal and mutated forms of the protein. We generated stable transfectants of full-length human endoglin in murine fibroblasts and engineered and expressed in bacteria several fragments of the extracellular domain. Relatively pure polypeptides were recovered with good yield from inclusion bodies and were tested by ELISA and Western blot; 11 monoclonal antibodies were shown to react specifically with the endoglin transfectants. Ten of these monoclonal antibodies reacted with the bacterial fragments, and their epitopes were assigned to 3 distinct regions of endoglin. Monoclonal antibodies P3D1, TEC4 and GRE reacted with the N-terminal region of 204 amino acids encoded by exons 1 to 5. Monoclonal antibodies P4A4, 44G4, E-9, MAEND3 and PN-E2 all bound to a region of 54 amino acids encoded mostly by exon 7. Monoclonal antibodies CLE4 and RMAC8 reacted with the C-terminal region of the extracellular domain, coded for by exons 8 to 12. Knowing the localization of these epitopes will facilitate the structural and functional analysis of normal and mutated forms of endoglin. PMID- 9331951 TI - High-resolution HLA typing for the DRB3/4/5 genes by sequence-based typing. AB - The high degree of polymorphism of the HLA genes at the nucleotide sequence level has proven sequence-based typing a major typing strategy. For DRB1 the allelic variability is predominantly present in the second exon and by DNA sequencing of exon 2 all hitherto known DRB1 alleles can be detected. For the associated genes DRB3, DRB4 and DRB5 the situation is slightly different. Allelic differences are not limited to exon 2 and the sequence of exon 3 and sometimes exon 4 is needed for complete subtyping. Oligonucleotides to amplify the exons needed for subtyping of DRB3, DRB4 and DRB5 were designed. Gene-specific products were generated to make simultaneous detection of alleles in heterozygous combinations possible. In this way 238 individuals were fully typed for their DRB3, 4 and 5 subtypes. Additional samples were typed for only one of the genes. All samples had been previously typed by PCR-SSP. Concordant typing results were obtained for all individuals tested. The DRB3 alleles typed for included *0101, *0201, *0202 and *0301, for DRB4 they were *01011, *0102 and *0103 and for DRB5 *0101, *0102, *0103, *0105, *0201, *0202 and *0203. All alleles were easily detected by the protocol described except for DRB5*0201. Sequencing of exon 3 and 4 of the DRB5*0201 allele showed this allele to be a sequencing error and the sequences obtained were identical to the exon 2, 3 and 4 sequences of DRB5*0202. Two new alleles were identified in the samples studied, DRB4*0105 and DRB3*0207. Sequence based typing has been recognized as a valuable tool for HLA typing of DRB1, DQB1 and DPB1 since several years. It is shown to be a superior typing method as well in the detection of the different DRB3, 4 and 5 subtypes. PMID- 9331952 TI - A three-step allele-level DRB1-DRB3-DRB4-DRB5 genotyping assay using polymerase chain reaction with immobilized sequence-specific oligoprobes. AB - A three-step reverse hybridization assay for allele level-resolution DRB1-DRB3 DRB4-DRB5 genotyping is described. Samples are initially amplified using a generic primer pair for all DRB1-DRB3-DRB4-DRB5 alleles and PCR products are hybridized to generic typing membranes. An intermediate-resolution level genotyping is obtained at this point. Depending on the phenotype, samples are then subjected to a DR1, DR2, DR4, DR52A, DRB3 and/or DRB5 type-specific amplification and hybridization. A third step, involving sequence-specific PCR followed by type-specific hybridization, is only performed to solve certain DR4 and DR52A heterozygous combinations. The assay allows 100% allele level resolution DRB genotyping. Hybridization membranes contain panels of SSO probes that were optimized to all react specifically under identical stringency conditions. A computer program was written to assist in analysis of the hybridization patterns. The assay was throughly evaluated and has been used to type over 10,000 donors from the Canadian Unrelated Bone Marrow Donor Registry (UBMDR) at allele level-resolution. This method proved to be flexible, easy to update for newly described alleles, easy to perform, fast, and safe. It is also reliable and specific, as 9 novel DRB alleles so far have been detected as aberrant hybridization patterns. PMID- 9331953 TI - Tumor necrosis factor locus polymorphisms in rheumatoid arthritis. AB - We examined six polymorphic elements in the tumor necrosis factor (TNF) locus and determined their allelic distribution in 98 Caucasian rheumatoid arthritis patients in comparison with 91 ethnically-matched controls. Polymorphic elements at four biallelic sites were distributed similarly between patients and controls, irrespective of the presence or absence of DR4. Differences were observed between the two groups at the TNFa and TNFe loci, but these were consistent with extended MHC haplotypes known to be present in rheumatoid arthritis patients. Therefore, this study suggests that there is little, if any, independent contribution of the TNF locus to the genetic background for rheumatoid arthritis susceptibility. PMID- 9331954 TI - Characterization of a novel HLA-B allele, B*0804, in a Norwegian family. PMID- 9331956 TI - Different trends in cardiovascular mortality and food consumption in Austria and in former Czechoslovakia. AB - The life expectancy of the male and female population immediately after World War II was similar in the former Czechoslovakia and in Austria. However, the situation has changed radically over the past 30 years. In 1991 the life expectancy in Czechoslovakia was 4.9 years less for males and 3.6 years less for females than in Austria due to the two-fold higher premature mortality from cardiovascular diseases and some further causes. The reasons for the 200% difference in cardiovascular mortality between Austria and Czechoslovakia are unknown. The prevalence of traditional risk factors (i.e. smoking, hypertension and hypercholesterolemia) was only moderately different in both countries. On the other hand, very different trends in food consumption were observed: in Austria the consumption of milk and milk products, vegetable oils, fruits, some kinds of vegetables and nuts was significantly higher, whereas the consumption of butter, sugar, eggs, cereals and concentrated spirits was lower. The consumption of pork, beef, poultry and fish was very similar in the two countries. The intake of antioxidant vitamins and probably also of folic acid, which affects the level of homocysteine, was lower in Czechoslovakia. The psychosocial load in totalitarian Czechoslovakia was undoubtedly higher. Epidemiological research of Central European populations living on both sides of the former Iron Curtain might bring new information on the role of less known non-traditional risk factors (i.e., psychosocial stress, nutritional factors such as chronic deficiency of antioxidant vitamins and a low intake of folic acid) in the etiology of cardiovascular diseases. PMID- 9331957 TI - 25-hydroxyvitamin D absorption in patients with Crohn's disease and with pancreatic insufficiency. AB - Vitamin D malabsorption could be one possible reason for the high prevalence of vitamin D deficiency and osteopenia in patients with Crohn's disease (CD) and pancreatic insufficiency (PI). Hence, we performed a modified 25-hydroxyvitamin D (25-OHD) absorption test Stamp in 15 healthy controls, 31 patients with CD and 10 patients with PI. Serum 25-OHD levels were measured before, and 2, 4, 8, and 24 hours after oral administration of 5 micrograms 25-OHD/kg body weight. Basal 25 OHD levels were below the normal range of 12-36 ng/ml in 68% of patients with CD (median: 10; interquartile range: 4-12 ng/ml) and 70% of patients with PI (median: 3; interquartile range: 2-14 ng/ml). Peak levels were reached at 4 or 8 hours after ingestion of 25-OHD. Three patients with CD (10%) and 5 patients with PI (50%) showed decreased 25-OHD absorption. 25-OHD levels normalized in all but two patients with PI after 24 hours. Pattern of involvement or previous resections did not show a significant influence on 25-OHD absorption. Vitamin D malabsorption may be one reason for vitamin D deficiency in many patients with PI, but there is little evidence of vitamin D malabsorption in patients with CD. Oral 25-OHD administration seems to be a useful therapeutic alternative to native vitamin D in patients with possible malabsorption and vitamin D deficiency. PMID- 9331955 TI - Complete sequence of HLA-B*1522: a class I allele that types by serology as HLA B35. PMID- 9331958 TI - Phase II trial of gemcitabine in advanced non-small-cell lung cancer. AB - Gemcitabine has shown activity in different solid tumors. In the present study we have evaluated its efficacy in 32 patients with advanced non-small-cell lung cancer in a phase II trial. Gemcitabine (1250 mg/m2) was given intravenously as a 30-minute infusion on days 1, 8 and 15. Cycles were repeated every 4 weeks. Twenty-nine patients were evaluable for response and all patients for toxicity. Partial remissions and stable disease were seen in 4 (14%) and 13 (45%) patients, respectively. Improvement of symptoms occurred in 54% of the patients. Side effects were mild and included predominantly leukopenia and thrombocytopenia. In conclusion, gemcitabine is active and well tolerated in patients with advanced non-small-cell lung cancer. PMID- 9331959 TI - Factor IX of the blood coagulation system: a review. AB - Factor IX is a factor of the blood coagulation system. Its activation occurs on the surface of phospholipid membranes. It can be activated by the factor VIIa-TF (tissue factor)-Ca2+ complex via an extrinsic pathway and by factor XIa in the presence of Ca2+ via the intrinsic pathway of blood coagulation system activation. The activated factor IXa is a serine proteinase. The main function of the activated factor IXa in complex with factor VIIIa and phospholipids in presence of Ca2+ consists of the activation of factor X. Factor IX is synthesized in the liver and is subject to a number of posttranslational modifications including gamma-carboxylation, beta-hydroxylation, and glycosylation. It forms a subgroup of vitamin K-dependent plasma proteins including factors VII and X and protein C characterized by identical domain structures having high levels of homology. Factor IX consists of an NH2-terminal Gla domain, two epidermal growth factor (EGF)-like domains, and a C-terminal domain containing Ser in its active site. Factor IX deficiency in human plasma results in the disease known as hemophilia B. PMID- 9331960 TI - Tocopherol modulates the effects of A23187, verapamil, and phorbol myristate acetate on RNA-polymerase activity of isolated rat liver nuclei. AB - Preincubation of rat liver nuclei with tocopherol decreased inhibition of RNA polymerase activity of isolated rat liver nuclei by A23187, verapamil, and phorbol myristate acetate (PMA). In nuclei of vitamin E-deficient rats, A23187, verapamil, and PMA did not inhibit label incorporation into RNA with and without tocopherol. A23187, verapamil, and PMA did not inhibit the activity of nuclei treated with 1% Triton X-100; tocopherol activated RNA synthesis but co administration of A23187 or verapamil and tocopherol had no effect and the combination of tocopherol and PMA inhibited RNA-polymerase activity by 25%. The effect of the combination of verapamil and PMA in the presence of free tocopherol was different from the effect of these compounds assayed in the presence of the complex of tocopherol with tocopherol-binding protein. PMID- 9331962 TI - Polypeptide composition of submitochondrial fractions of normal liver tissue and Zajdela hepatoma. AB - The polypeptide composition of liver and Zajdela hepatoma mitochondria is compared. Polypeptides of mitochondria and submitochondrial fractions (the outer and the inner mitochondrial membranes, the intermembrane and the matrix spaces of mitochondria) were separated by electrophoresis in polyacrylamide gel. The percentage content of each polypeptide was evaluated after scanning gels using a differential spectrophotometer-densitometer. Significant changes of several proteins of the hepatoma mitochondria and submitochondrial fractions as compared with normal liver preparations have been found. PMID- 9331961 TI - Reconstitution of the intercellular transfer pathway of the peptide moiety of ceruloplasmin in mammals. AB - According to rocket immunoelectrophoresis, the antigenic properties of the ceruloplasmin (Cp) receptor present on the surface of human fibroblasts are similar to those of the Cp receptor of the erythrocyte plasma membrane. Using antibodies to Cp receptor, it was demonstrated that Cp binds to the surface of fibroblasts only via the Cp receptor. Ceruloplasmin was labelled with radioactive iodine and its binding to cultured human HT-1080 fibroblasts was studied; at saturating concentrations [125I]Cp interacts with cell surface of fibroblasts (but not hepatocytes) with high affinity (Kd = 80 nM). Subsequent to specific binding fibroblasts absorb [125I]Cp and in about 120-150 min start to release it into the incubation medium. Two fractions of released Cp are clearly detected by non-denaturing polyacrylamide gel electrophoresis. The relative mobility of one of these fractions corresponds to apo-Cp and the other has lower mobility than native Cp. The molecular weight of released [125I]Cp is changed insignificantly. Released Cp does not bind again to the fibroblast surface but is bound, absorbed, degraded, and secreted by hepatocytes. The molecular mechanism of cell-specific transfer of Cp in the human body is discussed and possible functions of this mechanism in copper absorption, metabolism, and excretion in mammals are considered. PMID- 9331963 TI - Identification of Bacillus sp. FTU strain and the study of the caa3-type oxidase homology. AB - The culture, morphology, and genome of alkalo- and halotolerant bacterial strain Bacillus sp. FTU were characterized; the strain is compared to other representatives of genus Bacillus. The DNA-DNA hybridization data indicate that the strains of Bacillus halodurans DSM 497 and DSM 2513 and Bacillus sp. FTU belong to the same species. Bacillus sp. FTU can be renamed to Bacillus halodurans FTU. The N-terminal amino acid fragments of the subunits I and II of the terminal caa3-type cytochrome c oxidase of B. halodurans FTU were sequenced. The N-terminal fragments of this enzyme and of the caa3-type oxidase of alkalophilic Bacillus firmus OF4 are highly homologous (homology of subunits I and II is over 90 and over 96%, respectively). Such high homology of the terminal oxidases of these bacteria might be due to their alkaline medium. PMID- 9331964 TI - Effect of cobalt chloride on content of lipids and lipoproteins in serum and liver of rats. AB - Lipids and the composition of lipoproteins in blood serum and liver cytosol, total lipid, and phospholipid contents in liver subcellular fractions and the spectrum of microsomal liver lipids were studied in male Wistar rats after a single injection of cobalt chloride. Virtually all lipid and lipoprotein fractions in blood and liver were increased and lipoprotein composition was changes. The lipid composition of liver microsomes did not change under these conditions. Thus, microsomal membranes are stable under developing oxidative stress. PMID- 9331965 TI - Isolation of a strain overproducing endonuclease Eco29kI: purification and characterization of the homogeneous enzyme. AB - The physical map of the plasmid pSACII1 carrying the genes of restriction modification system Eco29kI (isoschizomer of SacII) was determined. The cloning of the Eco29kI endonuclease and methylase genes into the plasmid vector pUC129 produced recombinant strain Escherichia coli K802[pECO29A15] with Eco29kI synthesis level about 100 times higher than in the parent strain. The restriction endonuclease was purified from Escherichia coli K802 [pECO29A15] cells to near homogeneity using column chromatography sequentially on phosphocellulose, hydroxyapatite, and heparin-Sepharose and rechromatography on phosphocellulose. Biochemical characterization of the homogeneous R Eco29kI is given. The enzyme has molecular mass 24.5 kD and is present in the solution as a monomer. PMID- 9331966 TI - ADE6 gene of Saccharomyces cerevisiae yeast encoding formylglycinamidine ribonucleotide synthetase. Cloning, sequencing, and analysis. AB - The ADE6 gene of Saccharomyces cerevisiae yeast encoding the enzyme formylglycinamidine-ribonucleotide (FGAM)-synthetase of de novo synthesis of purine nucleotides was cloned and sequenced. The gene encodes a protein consisting of 1358 amino acids. The flanking regions of 1208 (5') and 728 bp (3') were also sequenced. The nucleotide motif TGACTC inherent to the promotor regions of other purine genes of yeast was located (-276 bp) in the 5'-region of the gene. The amino acid sequence of the yeast FGAM-synthetase was found to contain repeats (Leu430-Ala620 and Pro810-Ile1000). Repeats of similar patterns of conserved amino acids were also detected in the structure of all other FGAM synthetases. A homology of FGAM-synthetases with certain proteins of viruses from the Herpesviridae family was found. PMID- 9331967 TI - Inhibition of solubilized superoxide dismutase co-immobilized with catalase by the polydisulfide of gallic acid. AB - The catalytic activity of superoxide dismutase (SOD) and its conjugates with catalase and polymer peroxidase (p-peroxidase) obtained during covalent binding of enzymes with aldehyde dextrans was indirectly characterized by inhibition of adrenaline autoxidation in 0.1 M bicarbonate buffer, pH 10.2, and in microemulsion of 0.1 M aerosol OT (AOT) and Triton X-45 in octane containing 15% aqueous phase. The polydisulfide of gallic acid (PDGA) effectively inhibited SOD and its conjugates by a mixed mechanism. The inhibition constants Ki for SOD and its conjugate (SOD-catalase)mic in 0.1 M bicarbonate buffer, pH 10.2, were 0.1 and 0.25 microM, respectively. Autoxidation of PDGA by molecular oxygen in alkaline media (pH 10.2) influenced its inhibitory properties in buffer solution and microemulsion of AOT and Triton X-45 in octane. The radical chain mechanism of co-oxidation of adrenaline and PDGA apparently includes the anion radical O2-. as a coupling agent which propagated the chain. PMID- 9331968 TI - Comparison of Cu,Zn-superoxide dismutases of clawed frogs of the genus Xenopus. AB - Isoenzyme profiles and thermal stability of Cu,Zn-superoxide dismutase (Cu,Zn SOD) of clawed frogs of genus Xenopus (X. laevis and X. borealis) were compared. Earlier the presence of an unusually thermolabile mutant Cu,Zn-SOD as well as normal enzyme was shown in X. laevis. Having confirmed the data, we further show that X. borealis contains only the thermolabile Cu,Zn-SOD. This suggested that a mutation in the Cu,Zn-SOD gene and genome duplication have occurred in the immediate predecessor of X. laevis. Later on the parent Cu,Zn-SOD gene was lost, leaving only the mutated form of Cu,Zn-SOD in X. borealis. PMID- 9331969 TI - Synthesis and some aspects of topogenesis of bovine cytochrome P450scc in yeast. AB - A plasmid for effective expression of recombinant DNA encoding a hybrid protein composed of the N-terminal targeting presequence of subunit IV of yeast cytochrome c oxidase preceding the mature polypeptide chain of bovine cytochrome P450scc (pCoxIV-CYP11A1) in yeast has been constructed. It has been shown that this protein, when synthesized in yeast cells, in imported into mitochondria and undergoes proteolytic processing, thus yielding a product of molecular mass corresponding to that of mature cytochrome P450scc. However, only insignificant portion of the imported protein proves to be inserted into the inner membrane of heterologous mitochondria. The membrane-bound cytochrome P450scc exhibits cholesterol hydroxylase activity towards 22R-hydroxycholesterol in the presence of exogenous adrenodoxin and adrenodoxin reductase. This fact indicates that the foreign protein is correctly folded and oriented in the membrane. Thus, insertion into the inner membrane is a limiting step of the pCoxIV-CYP11A1 topogenesis in yeast cells, whereas its import into mitochondria and proteolytic processing proceed without significant impediments. PMID- 9331970 TI - Enzymatic oxidation of beta-apo-8'-carotenol to beta-apo-14'-carotenal by an enzyme different from beta-carotene-15,15'-dioxygenase. AB - Extracts of rat and rabbit intestinal mucosa were fractionated with ammonium sulfate. The precipitate contained beta-apocarotenoid-14',13'-dioxygenase (ADO) activity. beta-Apo-14'-carotenal was found to be the product of enzymatic cleavage of beta-apo-8'-carotenol. Activities of ADO and beta-carotene-15,15' dioxygenase (CDO) were detected in the presence of thiols and were inactivated by 1,10-phenanthroline. Optimal pH values were 7.0 for ADO and 8.0 for CDO. Heating at 52 degrees C inhibited ADO by 70% and produced no effect on CDO. ADO activity was maximal in the presence of sodium cholate or 3-[(3-cholamidopropyl)-dimethyl ammonio]-1-propanesulfonate (CHAPS). Sodium dodecylsulfate was required for maximal CDO activity. Proteins with ADO activity were not retained by phenyl Sepharose CL-4B. CDO activity was eluted from columns only in the presence of detergents. The data suggest that the enzymes that catalyze the oxidative cleavage of beta-carotene to yield retinal are different from those that cleave beta-apo-8'-carotenol to yield beta-apo-14'-carotenal. PMID- 9331971 TI - Mechanism of induced resistance to protein inhibitors of proliferation. AB - A 12 kD protein (IHP-12) was isolated from the cytosol of liver cells; IHP-12 inhibited proliferation of the liver cells by 50% when injected at 2.5 mg/100 g animal weight. Repeated injections of IHP-12 induced resistance of the liver cells to its anti-proliferative effect. After intraperitoneal injection, IHP-12 is localized in the cytosol and to a lesser extent, in cell nuclei. IHP-12 is degraded in the cytosol but in nuclei it retained its native structure for at least 8 h. The mechanism of induced resistance to IHP-12 was not related to a change in cell binding of IHP-12, its distribution in cellular compartments, and intracellular metabolism. The induced resistance may be associated with a change in the direction of IHP-12 activity, i.e., change in its function in the animal. PMID- 9331973 TI - Bioassay of human granulocyte colony-stimulating factor using human promyelocytic HL-60 cells. AB - A new method for an assay of human granulocyte colony-stimulating factor (hG-CSF) has been developed using human promyelocytic HL-60 cells. The proliferation of HL 60 cells had been suppressed by the addition of dimethyl sulfoxide (DMSO) or retinoic acid (RA). These differentiating agent-treated HL-60 cells exhibited an increase in their number in response to recombinant hG-CSF (rhG-CSF). Neither dibutyl-cAMP nor interferon-gamma (IFN-gamma)-treated HL-60 cells, however, showed an increase in their number in response to rhG-CSF. The proliferation rate of DMSO-pretreated HL-60 cells was linearly increased from 0.3 to 10 ng/ml of rhG CSF. L-Value of HL-60 cells assay was 0.027 +/- 0.012. The activities of non glycosylated rhG-CSF produced by Escherichia coli and glycosylated rhG-CSF produced by chinese hamster ovary (CHO) cells were compared using DMSO-treated HL 60 cells; no significant difference between them. DMSO-treated HL-60 cells also responded to interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF), but did not respond to erythropoietin or macrophage colony stimulating factor, suggesting that the responsiveness of these cells to growth factor is restricted to myelogenic cytokines. In conclusion, DMSO- or RA-treated HL-60 cells are useful for the measurement of bioactivity of hG-CSF. PMID- 9331974 TI - Production and characterization of monoclonal antibodies against two haptenic derivatives of 1 alpha,25-dihydroxyvitamin D3 conjugated with bovine serum albumin through the C-3 or C-24 position. AB - To develop the immunochemical methods for determining 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] in clinical samples, a variety of monoclonal anti-1,25(OH)2D3 antibodies have been generated. Two kinds of hapten-carrier conjugates, 25 hydroxyvitamin D3 3-hemisuccinate (hapten 3-HS) and 1 alpha-hydroxy-25,26,27 trinorvitamin D3 24-oic acid (hapten 24-OA) conjugated with bovine serum albumin, were used for immunization. Spleen cells from SD rats or BALB/c mice, each immunized with the conjugate of hapten 3-HS or 24-OA, were fused with P3/NS1/1 Ag4-1 myeloma cells. After screening by ELISA employing beta-galactosidase labeled haptens, seven kinds of hybridomas secreting anti-1,25(OH)2D3 antibodies were established. Binding characteristics of these antibodies (Ka 0.73-20 x 10(9) M-1) were investigated by an RIA using tritium-labeled 1,25(OH)2D3. The data suggested that the rat monoclonal antibody 3R-1 derived from the hapten 3-HS and the mouse monoclonal antibody 24M-3 from the hapten 24-OA would be available for developing practical analytical systems. PMID- 9331975 TI - The effects of Hange-shashin-to on the content of prostaglandin E2 and water absorption in the large intestine of rats. AB - The effects of Hange-shashin-to (TJ-14) were examined regarding the amount of prostaglandin E2 (PGE2) and the water absorbing capacity in the large intestine of rats. Repeated oral administration of TJ-14 at doses of 125 to 1000 mg/kg revealed a significant decrease in PGE2 content in the colonic mucosa and also the promotion of colonic water absorption in a dose dependent manner. However, there was no remarkable influence on the concentrations of aldosterone and electrolytes in the serum, even at 1000 mg/kg. From these results, it was considered that some of the anti-diarrheal effects of TJ-14 might be based on a repression of the increase in the amount of PGE2 as well as promotion of the water absorbing capacity of the large intestine. Moreover, it was also suggested that it is possible, by the application of TJ-14, to prevent the loss of water content caused by diarrhea. PMID- 9331976 TI - Selective cytotoxicity of piperine on cultured rat hippocampal neurons in comparison with cultured astrocytes: the possible involvement of lipid peroxidation. AB - The present study investigated the selective cytotoxicity of piperine on cultured brain cells by using lactate dehydrogenase release, 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium reduction, and protein content assay. Exposure to piperine induced marked injuries on cultured hippocampal neurons whereas it induced only a marginal toxicity on cultured astrocytes. The mechanism of cytotoxicity in hippocampal neurons was studied by pharmacological methods. The membrane-permeable free radical scavengers, D-alpha-tocopherol and trolox, protected neurons from piperine toxicity whereas membrane-impermeable agents, superoxide dismutase and catalase, were ineffective. A lipoxygenase inhibitor, nordihydroguaiaretic acid, but not a cyclo-oxygenase inhibitor, indomethacin, attenuated piperine toxicity. We provide the first evidence that piperine induces selective neurotoxicity in vitro. Although the exact mechanism is still unclear, pharmacological evidence indicates the possible involvement of lipid peroxidation and the generation of free radicals, at least partly, through an arachidonate lipoxygenase pathway. PMID- 9331977 TI - Regulation of aldosterone synthase cytochrome P450 (CYP11B2) and 11 beta hydroxylase cytochrome P450 (CYP11B1) expression in rat adrenal zona glomerulosa cells by low sodium diet and angiotensin II receptor antagonists. AB - Changes in the mRNA levels for aldosterone synthase cytochrome P450 (CYP11B2 or P450aldo) and 11 beta hydroxylase cytochrome P450 (CYP11B1 or P45011 beta) in rat adrenal glands were studied in response to angiotensin II type 1 (AT1) and type 2 (AT2) receptor antagonists. CYP11B1 and CYP11B2 genes were highly homologous (88.5%) in their nucleotide sequences of the amino acid coding regions. Reverse transcription-polymerase chain reactions (RT-PCR) which are capable of discriminating between rat CYP11B1 and CYP11B2, were performed with specific primers for each P450. Upon sodium restriction (5 mmol Na(+)/kg of diet) of rats for 14d, the amount of the CYP11B2 mRNA in the adrenal glands was increased 8.5 fold compared to that from the rats fed a normal diet (225 mmol Na(+)/kg of diet), whereas no significant change in the CYP11B1 mRNA was observed after the dietary sodium restriction. As shown by an immunoblot analysis, the adrenal capsule portions (mainly zona glomerulosa) of the rats kept on the low Na diet for 14d expressed significantly higher levels of both CYP11B2 and CYP11B1, and contained a significantly higher amount of CYP11B2 than those from the rats fed by normal diet. The activities of the CYP11B2 enzyme were also found to be increased by about 8-fold on day 14. In concert with these alterations, the plasma aldosterone concentration (PAC) increased. However, when the specific AT1 antagonist E4177 was given to rats maintained on the low Na diet, the amount and activity of CYP11B2, as well as the PAC, were suppressed. In contrast, the increase in CYP11B2 induced by the low Na diet was not affected by the AT2 specific antagonist PD123177. These results indicate that the aldosterone synthase cytochrome P450 (CYP11B2) is an ultimate target of the regulation of aldosterone biosynthesis by an AT1 receptor antagonist. PMID- 9331978 TI - Hair analysis for drugs of abuse. XVIII. 3,4-Methylenedioxymethamphetamine (MDMA) disposition in hair roots and use in identification of acute MDMA poisoning. AB - Disposition of 3,4-methylenedioxymethamphetamine (MDMA) in hair roots was studied using rats and the hair root samples were evaluated to prove acute MDMA poisoning. The back hair of male pigmented hairy rats (n = 6) was shaved and 5 d later the animals were intraperitoneally administered with acute poisonous doses (20, 40, 60, 80 and 100 mg/kg) of MDMA. Roots of the hairs were then plucked out with a hair nipper 5 min and, 0.5, 1, 2, 6 and 24 h after injection. The hair root samples were, directly or after being washed with detergent, extracted with methanol-5 N HCl (20:1) under ultrasonication in ice-cold water for 4 h. After filtration and evaporation, the residue was derivatized with pentafluoropropionic anhydride and analyzed by GC/MS. From all samples including the 5 min sample, MDMA was detected at high concentrations (up to 156 ng/mg) accompanied by 3,4 methylenedioxyamphetamine (MDA). Some of the animals died within 2 h after administration, but in the surviving rats the MDMA concentrations in the hair roots increased up to 6 h and then slowly decreased until 24 h. The remaining MDMA after washing apparently increased from 13-31% at 0.5 h to 51-83% at 24 h in the surviving rats. These facts show that most of drugs in the hair roots are not yet immobilized in the early stage and are thereafter gradually incorporated into the hair shaft. Increase of the MDMA concentration stopped soon after death of the animal, probably due to the cessation of hair growth. Although the ratios of MDA/MDMA steadily increased over time, those after death plateaued, probably due to the cessation of metabolism after death. It was clearly shown that MDMA is more quickly incorporated into and more firmly retained in hair than methamphetamine (MA) by comparing their disposition in hair roots. PMID- 9331979 TI - Biochemical characterization of glycyrrhizin as an effective inhibitor for hyaluronidases from bovine testis. AB - The inhibitory effects of several anti-inflammatory agents, including glycyrrhizin (GL), on the activities of hyaluronidases (HAses) purified from bovine testes and Streptomyces were investigated in vitro. It was found that (i) GL inhibits the activity of HAse (p55) from bovine testes in a dose-dependent manner, but does not affect HAse from Streptomyces; (ii) GL was the most effective of the compounds tested on bovine testis HAse activity (50% inhibition with approx. 3 microM GL); and (iii) glycyrrhetinic acid (GA), a derivative (oGA) of GA and diglucuronic acid had no detectable effects on HAse activity at 9.0 microM. The GL-induced inhibition of HAse activity is uncompetitive for its substrates. Data are provided to support the contentions that (i) bovine testis HAse (p55) is a GL-binding protein; and (ii) GL acts as a potent inhibitor of HAse in vitro. PMID- 9331980 TI - Effects of alkaloids from Aconitum yesoense var. macroyesoense on cutaneous blood flow in mice. AB - Nine alkaloid constituents in the root of Aconitum yesoense var. macroyesoense, as well as three acetylated derivatives, were examined for their peripheral vaso activities by measuring laser-flowmetrically the cutaneous blood flow in the hind foot of mice after intravenous administration. The major constitutive delcosine (1), 14-acetyldelcosine (2) and lucidusculine (3), respectively, had little or very mild vaso-activity. Kobusine (4) and pseudokobusine (5) and three minor constituents, luciculine (6), 1-acetylluciculine (7) and dehydroluciculine (8), together exhibited a rapid increase in blood flow reaching a peak with a magnitude almost equal to that produced by hydralazine, when administered intravenously at the same dosage level of 20 mg/kg. Among them, 4 was characterized by successive reversal of the increase to a decrease in blood flow, while 7 produced a flow with a more delayed peak time. Dehydrolucidusculine (9) exhibited a transient decrease in blood flow prior to occurrence of the increase, as did papaverine. Consequently, it is assumed that the alkaloids, especially those of the C20-diterpenoid type, in the root of this Aconitum plant have peripherally vaso-dilating activities to varying degrees in mice, probably due to their direct action on the cutaneous microvasculature in a similar fashion to that shown by hydralazine. The laser blood flowmetric method would be useful as an in vivo means of qualitative as well as quantitative screening of chemically modified derivatives of peripherally vasoactive agents in mice. PMID- 9331981 TI - Study on baths with crude drug. III. The effect of ligustici chuanxiong rhizoma extract on the percutaneous absorption of some natural compounds. AB - To investigate the permeability of natural compounds through hairless mouse skin, compounds having a range of lipophilicity, i.e., ginsenoside-Re (1), baicalein (2), glycyrrhizin (3), baicalein (4), wogonin (5), honokiol (6), magnolol (7), bergapten (8), shikonin (9) and sinomenine (10) were used. These compounds permeated through the skin a little, however, they were generally accumulated into the skin. The uptake amount into the skin of each compound related to their lipophilicities in the in vitro experiment. Furthermore, Ligustici Chuanxiong Rhizoma (Senkyu) ether extract (SEE) enhanced their permeability into the skin; especially, it exhibited an effect on the skin permeability of moderately lipophilic compounds such as 4, 8. The effect of SEE in vivo was similar to that obtained in the in vitro experiment. From these results, it was clarified that natural compounds having high lipophilicity sufficiently permeated into the hairless mouse skin owing to their accumulative property, and SEE enhanced the permeability of the moderately lipophilic compounds into the skin. PMID- 9331982 TI - Preventive effects of saponins from puerariae radix (the root of Pueraria lobata Ohwi) on in vitro immunological injury of rat primary hepatocyte cultures. AB - The preventive effects of saponins from Puerariae Radix toward in vitro immunological liver injury using an antiserum against the rat liver plasma membranes on primary cultured rat hepatocytes were studied. Crude saponin from Puerariae Radix inhibited the elevation of alanine aminotransferase (ALT) activity at the dose of 90 micrograms/ml. The inhibition was stronger than that of glycyrrhizin, which was a positive control drug. The representative saponins in this drug, soyasaponin I and kudzusaponin SA3, were also more effective than glycyrrhizin, although their effects were weaker than that of crude saponin at the lower doses (90, 200 micrograms/ml). At 500 micrograms/ml, kudzusaponin SA3 showed antihepatotoxic activity equal to that of crude saponin. PMID- 9331983 TI - Plasma pharmacokinetics of 7-ethyl-10-hydroxycamptothecin (SN-38) after intravenous administration of SN-38 and irinotecan (CPT-11) to rats. AB - We studied the plasma pharmacokinetics of the lactone form and the carboxylate form of 7-ethyl-10-hydroxycamptothecin (SN-38), the active metabolite of irinotecan (CPT-11), after intravenous bolus administrations of each form of SN 38 and of CPT-11 to rats. After the SN-38 lactone injection, SN-38 lactone was predominant at first, and then the lactone to the carboxylate concentration ratio (LC ratio) was maintained from 30 to 90 min after the injection. The carboxylate was predominant throughout the period after the carboxylate dosing. Model dependent analyses showed that the SN-38 lactone had greater plasma clearance and a greater distribution volume than the carboxylate. The CPT-11 administration resulted in a predominant plasma SN-38 lactone concentration. The contribution of the SN-38 lactone AUC to the total SN-38 AUC (57%) was independent of the dose of CPT-11. These results suggest that it is possible to estimate the SN-38 lactone concentration and AUC from the total SN-38 concentration without separate determination of the lactone and carboxylate. Our results showed that both SN-38 lactone and CPT-11 administration gave the predominant SN-38 lactone in plasma; however, only CPT-11 could sustain the lactone concentration at a high level, which is necessary for antitumor activity. PMID- 9331984 TI - Differential effects of buprenorphine and pentazocine on the regional cerebral metabolic rate for glucose in the conscious rat. AB - The effects of buprenorphine (BNP, 10-200 micrograms/kg, i.v.) and pentazocine (PTZ, 2.5-10 mg/kg, i.v.) on the regional cerebral metabolic rate for glucose (rCMRglc) were analyzed in nine anatomically discrete areas of the conscious rat brain by the simultaneous use of [14C]2-deoxyglucose, the glucose analogue that can be phosphorylated in the brain, and [3H]3-O-methylglucose, a nonmetabolizable glucose analogue. Originally, this method was developed by Gjedde and Diemer in the rat and in humans. The rCMRglc was significantly decreased by BNP (100 or 200 micrograms/kg) in most of the brain regions investigated, except the cerebellum. In contrast, PTZ (10 mg/kg) significantly increased rCMRglc in the cerebral cortex and medulla. In the cerebral cortex and medulla, the direction of the effect on rCMRglc was opposite for BNP (22% decrease at the dose of 200 micrograms/kg) and PTZ (22% increase at the dose of 10 mg/kg). These findings strongly suggest that the discrepancies between the marked effects of BNP (a partial mu agonist and kappa antagonist) and PTZ (a mu antagonist and kappa agonist) on rCMRglc reflect the selectivity of agonist action at the different types of opioid receptors, mu and kappa receptors, in the rat brain. PMID- 9331985 TI - Breaks in double-strand DNA by Cu(II) complexes of etoposide (VP-16) and its derivatives, as evaluated by S1 nuclease treatment. AB - Single-strand breaks (ssb) in double-strand (ds) DNA produced by hydroxyl radicals (.OH) generated by Cu(II) complexes of podophyllotoxin (PD)-related compounds were evaluated using S1 nuclease digestion. Cu(II) complexes of VP-16 (etoposide, 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta-D-glucopyra noside)), 4'-demethylepi-PD (DEPD), and syringic acid (SA) exhibited both ssb and ds breaks (dsb) in ColE1-HaeII and pBR322-BglI DNA fragments, in which the number of ssb was found to be more than three times and four times greater than that of dsb, respectively. Cytosine (C)-methylation of cytosine-guanine doublet (CpG) in pBR322-BglI DNA inhibited both ssb and dsb within DNA segments by .OH generated by the Cu(II) complexes. PMID- 9331986 TI - Nitric oxide synthesis in murine peritoneal macrophages by fungal beta-glucans. AB - Fungal beta-glucans have abilities to induce NO (nitric oxide) synthesis by macrophages in vivo, and the intensity of NO synthesis significantly varied dependent on the structure of beta-glucans. The molecular mechanism of NO synthesis by beta-glucans, however, has not been clarified in detail. To determine beta-glucan-mediated NO production, we used various beta-glucans (SPG OH, GRN; Grifolan, SSG, OL-2, ZYM; zymosan A and ZYC; zymocel), which could enhance NO synthesis in vivo, and stimulated peritoneal macrophages (MPs) in vitro in the presence of interferon-gamma (IFN-gamma). Using recombinant cytokines, a minimum concentration of the cytokines for NO induction was about 20 mg/ml in the presence of IFN-gamma under the experimental conditions. Of beta glucans tested, only SPG-OH and GRN produced high concentrations of IL-6 in the culture supernatants. SSG also induced NO synthesis in vitro, but concentrations of inflammatory cytokines were low even in the presence of IFN-gamma. In addition, there are some beta-glucans which could induce NO synthesis in vivo but not in vitro (OL-2, ZYM, ZYC). These findings suggested that NO productivity of beta-glucans in vivo is regulated by several mechanisms. PMID- 9331987 TI - Evaluation of the standardized disk diffusion and agar dilution antibiotic susceptibility test methods by using strains of Neisseria gonorrhoeae from Tucuman, Argentina. AB - At present, most Neisseria gonorrhoeae testing is done with beta-lactamase and agar dilution tests using common therapeutic agents. Generally, in bacteriological diagnosis laboratories in Argentina, study of antibiotic susceptibility of N. gonorrhoeae is based on beta-lactamase determination and agar dilution method using common therapeutic agents. The National Committee for Clinical Laboratory Standards (NCCLS) recently described a disk diffusion test that produces results similar to the reference agar dilution method for antibiotic susceptibility of N. gonorrhoeae. We obtained 57 gonococcal isolates from patients attending a clinic for sexually transmitted diseases in Tucuman, Argentina. Antibiotic susceptibility tests using agar dilution and disk diffusion techniques were compared. The established NCCLS interpretive criteria for both susceptibility methods appeared to be applicable to domestic gonococcal strains. The correlation between the minimum inhibitory concentration (MIC's) and the zones of inhibition was studied for penicillin, ampicillin, cefoxitin, spectinomycin, cefotaxime, cephaloridine, cephalexin, tetracycline, norfloxacin and kanamycin. Dispersion diagrams showed a high correlation between both methods, with a sensitivity of 89% and specificity of 91%. PMID- 9331988 TI - A simple quantitative measurement of mRNA of human beta-actin by reverse transcription competitive PCR with a compact digital camera. AB - Reverse transcription (RT) competitive polymerase chain reaction (PCR) is a sensitive and useful technique for the absolute quantitation of mRNA level. We developed a competitive PCR method with a compact digital camera DC 40 (Eastman Kodak Company, NY, U.S.A.) for the measurement of human beta-actin cDNA (mRNA), a reference gene in an analysis for target gene expression. The devised method, compared with currently used methods, was easy, safe, sensitive, and highly reproducible to assay an infinitesimal level of cDNA (mRNA). PMID- 9331989 TI - Immunosuppressive effects of gallic acid and chebulagic acid on CTL-mediated cytotoxicity. AB - Gallic acid (GA) and chebulagic acid (CA) were isolated from the extract of a herbal medicine, kashi (myrobalans: the fruit of Terminalia chebula) as active principles that blocked the cytotoxic T lymphocyte (CTL)-mediated cytotoxicity. GA and CA inhibited the killing activity of CD8+ CTL clone at IC50 values of 30 microM and 50 microM, respectively. Granule exocytosis in response to anti-CD3 stimulation was also blocked by GA and CA at the equivalent concentrations. PMID- 9331990 TI - Effects of astemizole on ventricular activation delay and RT intervals in a canine myocardial infarction model. AB - In order to clarify the arrhythmogenic effects of nonsedating antihistamines, we examined the effects of astemizole, a nonsedating antihistamine, on ventricular activation and RT intervals in a canine myocardial infarction model. Myocardial infarction was produced by two-stage ligation of the left anterior descending coronary artery in dogs. Seven days after ligation, bipolar electrodes were sutured on the ventricular surface of the infarcted and normal zones to apply an electrical stimulation or record the ventricular activation. An electrical stimulation with coupling intervals between 300 and 140 ms was applied on the ventricular surface of the normal zone during atrial pacing, and the ventricular activation delay was measured. The effect of astemizole on the RT interval was also determined during atrial pacing, sinus rhythm or after premature stimulation. The ventricular activation delay increased after astemizole at doses of 0.3 to 3 mg/kg in the infarcted and at 3 mg/kg in the normal zones, and the effect of astemizole was greater in the infarcted zone. Astemizole increased the RT interval in the normal zone to a greater extent at a long coupling interval. The increase in the RT interval was greater in the infarcted zone compared with that in the normal zone. In conclusion, astemizole increased the activation delay in the infarcted zone, probably through prolongation of the repolarization time, and its effect on the activation delay may be arrhythmogenic. PMID- 9331991 TI - Preference of Peyer's patches to jejunal epithelium for intestinal absorption of oligopeptides, tyrosylglycylglycine and D-kyotorphin. AB - The intestinal absorption of oligopeptides, peptidase-degradable tyrosylglycylglycine (TGG) and peptidase-resistant L-tyrosyl-D-arginine (D kyotorphin, D-KTP) across Peyer's patches (PP) in rabbit intestine were studied using an Ussing-type chamber and in situ closed perfusion methods. The clearance for the serosal appearance of intact TGG across PP by the Ussing-type chamber method was a little higher than that across the jejunal epithelium (JE). Meanwhile, the in situ closed perfusion experiment showed that the clearance for the plasma appearance of intact TGG across PP was about 10 times that of JE. Furthermore, it was shown that the clearance for the plasma appearance of D-KTP in PP was about twice that in JE by the in situ closed perfusion method, indicating that the membrane permeability of PP was higher than that of JE. Therefore, these results indicate that PP had less metabolic peptidase activity than JE, and that the PP was a suitable site for the intestinal absorption of oligopeptides, especially peptidase-degradable peptides. PMID- 9331992 TI - Effect of non-insulin dependent diabetes on cyclosporin A disposition in Goto Kakizaki (GK) rats. AB - We previously reported that the pharmacokinetics of cyclosporin A (CyA), particularly absorption, were altered in diabetic rats treated with streptozotocin. In the present study, the effect of diabetes on pharmacokinetics of CyA after intravenous and oral administration of CyA using the blood and lymph and the gastrointestinal transit were examined in Goto-Kakizaki (GK) rats, a genetic model of non-obese non-insulin-dependent diabetes mellitus (NIDDM), and compared to non-diabetic Wistar rats. Although the systemic and lymphatic availability after intravenous administration of CyA to the GK rats was not significantly different from those of the control Wistar rats, those availability after oral administration of CyA to GK rats was markedly reduced in comparison. These results suggest that the pharmacokinetics of CyA, particularly absorption, was altered in GK rats. Studies on the gastrointestinal transit in GK rats showed that the gastric emptying rate was lower than that of Wistar rats, suggesting that a change in gastrointestinal transit in GK rats may influence the absorption of CyA. The gastric emptying rate in GK rats altered not only the systemic availability but also the lymphatic availability, suggesting that the altered systemic availability may cause adverse effects and that altered lymphatic availability may influence the immunosuppressive effects. PMID- 9331993 TI - Neurotoxic study of H2 antagonists using Xenopus oocytes injected with mouse brain mRNA. AB - To clarify the dominant mechanism for the convulsant activity of H2 antagonists, the effects of an H2 antagonist, cimetidine, on membrane currents induced by various agonists were investigated. In Xenopus oocytes injected with mouse-brain mRNA, acetylcholine (ACh), serotonin (5-HT), gamma-aminobutyric acid (GABA), glycine (Gly), glutamic acid (Glu), kainic acid (KA), quisqualic acid (QA) and N methyl-D-aspartic acid (NMDA)-induced current responses were recorded under a voltage-clamp condition. Cimetidine inhibited GABA-induced currents in a concentration-dependent manner; however, the current responses induced by the other agonists were not modified. The IC50 of various H2 antagonists, famotidine, nizatidine, cimetidine and ranitidine, for GABA (10 microM)-induced current response were 66, 260, 450 and 980 microM, respectively. However, these values of cimetidine and ranitidine were 40-400 times higher than the reported brain and cerebrospinal fluid (CSF) concentration of H2 antagonists at the occurrence of a clonic convulsion in vivo. In conclusion, we observed an inhibitory effect of H2 antagonists on the GABA response; however, this inhibition of GABA-mediated neurotransmission may not be the dominant mechanism for H2 antagonist-induced clonic convulsion in vivo. PMID- 9331994 TI - Bactericidal activity against Vibrio cholerae of chemical products used in lemon production in Tucuman, Argentina. AB - The present research was set up to verify whether the chemical products used in lemon production (from cultivation until packaging) have a bactericidal or a bacteriostatic ability against Vibrio cholerae O1. The studied products were: copper oxychloride, benomil (a carbamate), active chlorine, sodium-o phenylphenoate, guazatine (a polyamine mixture), imazalil (an imidazole) and lemon peel. The latter was studied with and without treatment using the above mentioned chemicals. Different dilutions of these products were tried out with varying exposure times against the bacterium V. cholerae Serogroup O1, Biotype E1 Tor, Serotype Inaba. The concentrations of the microorganism ranged from 10(2) to 10(8) CFU ml-1, the latter one being considered an infectious dose. The following results were obtained: 1) active chlorine (chlorinated water) showed bactericidal activity at concentrations of 50, 100 and 200 ppm after 10 min of exposure time, 2) copper oxychloride, sodium-o-phenylphenoate, guazatine and imazalil showed bactericidal activity against V. cholerae at concentrations of 10(2) and 10(4) CFU ml-1, 3) due to the fact that during its cultivation the fruit is successively sprayed with several chemical products, it could be that the result of the successive treatments is superior to the result of a repeated treatment with each of the individual products. This consideration should be taken into account when evaluating the eventual protection of the lemon. PMID- 9331995 TI - Comparison of characterization among Bordetella calmodulin-like protein, bovine brain calmodulin and Escherichia coli acyl-carrier protein. AB - We previously reported that Bordetella calmodulin-like protein (CLP), like bovine brain calmodulin (CaM), enhances adenylate cyclase and phosphodiesterase activities. In this communication, antigenic and biochemical characters were compared between CLP and CaM. It was found that anti-CLP and anti-CaM sera obtained reacted with CaM and CLP, respectively. The amino acid composition of CLP was similar to CaM. The similarity was also found in an Escherichia coli acyl carrier protein (ACP), a Ca(2+)-binding protein. Though the amino acid sequence of N-terminus region of CLP has no significant homology with CaM, ACP has highly homology in its N-terminus similar to CaM. These results suggest that CLP might act as a Ca(2+)-binding protein, and/or an ACP during the activation of adenylate cyclase and phosphodiesterase. PMID- 9331996 TI - 3 beta,5 alpha-Dihydroxycholestan-6-one exists in human blood. AB - 3 beta,5 alpha-Dihydroxycholestan-6-one was detected in fresh human plasma at the concentration of 14-44 ng/mL. The oxysterol binds specifically to phorbol ester specific binding protein in vitro, and may be an endogenous ligand of the protein. PMID- 9331997 TI - Synthetic studies on condensed-azole derivatives. V. Synthesis and anti-asthmatic activities of omega-sulfamoylalkyloxy[1,2,4]triazolo[1,5-b]-pyridazines. AB - A series of novel ([1,2,4]triazolo[1,5-b]pyridazin-6-yl) oxyalkylsulfonamides was synthesized and evaluated for the ability to inhibit platelet activating factor (PAF)-induced bronchoconstriction in guinea pigs. The compounds bearing a gem dialkyl or a cycloalkylidene group at the 2 position of the sulfamoylpropyloxy group in the side chain and a methyl group at the 7 position were found to have potent activity. Among them, 2,2-diethyl-3-(7-methyl[1,2,4]- triazolo[1,5 b]pyridazin-6-yl)oxypropanesulfonamide (13) showed excellent anti-asthmatic activity. Compounds 13 may be of significant value in the treatment of asthma and other respiratory diseases. The structure-activity relationships in this series of compounds are discussed. PMID- 9331998 TI - Synthesis of novel succinamide derivatives having a 5,11-dihydro-6H-pyrido[2,3 b][1,4]benzodiazepin-6-one skeleton as potent and selective M2 muscarinic receptor antagonists. II. AB - A series of succinamide derivatives containing the 5,11-dihydro-6H-pyrido[2,3 b][1,4]benzodiazepin-6-one skeleton (6a-z) was prepared and evaluated for binding affinity to muscarinic receptors in vitro and for antagonism of bradycardia and salivation in vivo in comparison with AF-DX 116 (1a). Structure-activity relationships (SAR) studies in vitro indicated that the 4-(4-alkyl-1 piperazinyl)benzylamino moiety plays a crucial role in enhancing the affinity for M2 muscarinic receptors. Compound 6y, containing a 4-(4-isopropyl-1 piperazinyl)benzylmethylamino moiety, exhibited the highest affinity for M2 muscarinic receptors (pKi = 9.2), being 200 times as potent as 1a, and compound 6u, containing a 4-(4-ethyl-1-piperazinyl)benzylethylamino moiety, showed the highest selectivity for M2 over M3 muscarinic receptors (M3/M2 ratio = 320). Both 6y and 6u antagonized the oxotremorine-induced bradycardia in rats after intravenous or oral administration. Oral evaluation in conscious dogs showed that the efficacy for increasing the heart rate was at least 3-fold greater than that of 1a. PMID- 9331999 TI - Synthesis and antitumor activity of novel benzophenone derivatives. AB - Novel benzophenone derivatives were synthesized and screened for cytotoxic and antitumor activity. Friedel-Crafts condensation was employed to construct the benzophenone skeleton. Among the compounds synthesized, morpholino and thiomorpholino benzophenones 3a-d exhibited potent cytotoxic activity against P388 murine leukemia and PC-6 human lung carcinoma cells in vitro, and compounds 3a, 3c, and 3j, when administered intraperitoneally, showed significant antitumor activity against the malignant ascites caused by intraperitoneal inoculation of P388 cells in mice. PMID- 9332000 TI - Studies on anti-inflammatory agents. V. Synthesis and pharmacological properties of 3-(difluoromethyl)-1-(4-methoxyphenyl)-5- [4-(methylsulfinyl)phenyl]pyrazole and related compounds. AB - A series of novel 1,5-diphenylpyrazole derivatives bearing hydrophilic substituents was prepared. The anti-inflammatory and analgesic activities of these compounds were evaluated by using the adjuvant arthritis and Randall Selitto assays in rats, and the structure-activity relationships were studied. The optimal compound was 3-(difluoromethyl)-1-(4-methoxyphenyl)-5-[4 (methylsulfinyl)phenyl]pyraz ole (10) with oral ED50 values of 0.31 and 2.6 mg/kg on adjuvant-induced arthritis and carrageenin-induced foot edema, respectively. Compound 10 showed analgesic activities not only toward inflamed paw but also toward normal paw (ED30 = 0.55 and 1.8 mg/kg, respectively) in the Randall Selitto assay, and moreover, 10 was effective in the tail-pinch assay (ED50 = 21 mg/kg) similarly to morphine. The asymmetric synthesis and pharmacological properties of the enantiomers of 10 are also reported. PMID- 9332001 TI - Saikosaponin homologues from Clinopodium spp. The structures of clinoposaponins XII-XX. AB - Nine new saikosaponin homologues, called clinoposaponins XII-XX, were isolated from the aerial parts of Clinopodium vulgare, C. chinense and C. chinense var. parviflorum together with nine known saikosaponin homologues. On the basis of spectral and chemical evidence, the structures of clinoposaponins XII-XX were determined to be 3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl] saikogenin F, 3-O-beta-D-fucopyranosyl-21 beta- hydroxysaikogenin F, 3-O-[beta-D glucopyranosyl-(1--> 3)-beta-D-fucopyranosyl]-21 beta-hydroxysaikogenin F, 3-O [beta-D- glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]saikogenin F, 3-O-[beta-D glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl- (1-->3)]-beta-D-fucopyranosyl]-21 beta-hydroxysaikogenin F, 3-O-[beta-D-glucopyranosyl-(1-->2)-[beta-D glucopyranosyl- (1-->3)]-beta-D-fucopyranosyl]-23-oxosaikogenin E, 3-O- [beta-D glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->3)]-beta- D-fucopyranosyl]-16 ketosaikogenin F, 3-O-[beta-D-glucopyranosyl-(1--> 2)-[beta-D-glucopyranosyl-(1- >3)]-beta-D-fucopyranosyl]-30-hydroxysa ikogenin F, 3-O-[beta-D-glucopyranosyl-(1 ->2)-[beta-D-glucopyranosyl-(1-->3)] -beta-D-fucopyranosyl]-30-oxo-saikogenin F, respectively. The known saponins were assigned to be clinoposaponins III, V, IX, X, XI, buddlejasaponins I, IV, 3-O-beta-D-fucopyranosylsaikogenin F and saikosaponin a. PMID- 9332002 TI - Bioactive constituents of Chinese natural medicines. IV. Rhodiolae radix. (2).: On the histamine release inhibitors from the underground part of Rhodiola sacra (Prain ex Hamet) S. H. Fu (Crassulaceae): chemical structures of rhodiocyanoside D and sacranosides A and B. AB - The methanolic extract of the underground part of Rhodiola sacra (PRAIN ex HAMET) S. H. Fu was found to show inhibitory activity on the histamine release from rat peritoneal exudate cells induced by an antigen-antibody reaction. From the methanolic extract with the inhibitory activity on histamine release, a new cyanoglycoside called rhodiocyanoside D and two new monoterpene glycosides called sacranosides A and B were isolated, together with eight known compounds, rhodiocyanoside A, heterodendrin, lotaustralin, rhodioloside, 2-phenylethyl alpha L-arabinopyranosyl(1-->6)-beta-D-glucopyranoside, 2-methyl-3-buten-2-yl beta-D glucopyranoside, kenposide A, and rhodiooctanoside. The structures of new compounds were determined on the basis of chemical and physicochemical evidence, which included the synthesis of sacranoside A from (-)-myrtenol. All major chemical constituents from R. sacra inhibited the histamine release and, among them, lotaustralin and rhodiooctanoside were found to show potent inhibitory activity. PMID- 9332003 TI - Characterization and stability of various liposome-encapsulated enoxacin formulations. AB - The necessity for antibacterial agents with greater intracellular efficacy has led to the development of endocytosable drug carriers such as liposomes. Enoxacin was selected as a model drug incorporated in various liposome formulations as a therapeutic dosage form using the ethanol injection method and freeze-drying. Liposomal behavior after preparation and stability test was characterized by determining the physicochemical properties of enoxacin encapsulation percent, vesicle size and turbidity. The non-phospholipid formulation of stratum corneum liposomes showed the highest encapsulation efficiency after preparation among nine liposomal formulations. The addition of dissacharides in liposomes also enhanced the encapsulation of enoxacin due to the protection of phospholipid bilayers during the freeze-drying process. The liposomes with negatively charged component and dissacharides showed lower enoxacin leakage after five weeks of storage at 45 degrees C, suggesting these formulations have high stability in long-term storage. The negative liposomes showed a different behavior than others in their decrease of size and turbidity during storage, possibly due to high surface charges of the negative formulation. Cholesterol stabilized bilayers interacted with plasma and high density lipoprotein (HDL) retained enoxacin in the vesicles. Nevertheless, liposomes with cholesterol caused a hydrolysis problem after incubation with normal saline. The formulation with trehalose not only showed high stability in storage but also in plasma and HDL. This suggested trehalose was useful to incorporate with phospholipids to produce a highly encapsulated and stabilized liposomes of enoxacin. This study also demonstrated that thought is required in utilizing turbidity as a direct index of liposomal vesicle size. PMID- 9332004 TI - Tableting of coated particles. I. Small particle size chitosan as an agent protecting coating membrane from mechanical damage of compression force. AB - Formulation of sustained release tablets containing coated particles whose coating membrane is not damaged during compression was studied and several kinds of chitosan of different particle size were evaluated as protective agents for the membrane. Comparison was made with the dissolution rate of the coated particles. Ethylcellulose or ethylcellulose/hydroxypropylcellulose was chosen as a coating agent. When the coated particles were compressed with the small particle size chitosan (Marine Chito), the coating membrane was not ruptured, and the protective effect was not influenced by the compression pressure. Both the Eudragit RS-coated particles and the tablets manufactured by compressing the coated particles with Marine Chito were orally administered to dogs, and the plasma theophylline levels of the two dosage forms were compared to determine the drug release characteristics in the gastrointestinal tract. It was found that the plasma concentration-time curve of the tablets coincided with that of the coated particles, and the compressed tablet would disintegrate instantly and redisperse into many particles in the body after oral administration. PMID- 9332005 TI - Anti-tuberculosis activity of quassinoids. AB - In vitro evaluation of anti-tuberculosis activity was conducted for fifty-six quassinoids isolated in our laboratory from Simaroubaceous plants, Ailanthus altissima (= Aa, 10 compounds), Brucea antidysenterica (= Ba, 16 compounds), Picrasma ailanthoides (= Pa, 14 compounds), and Brucea javanica (= Bj, 16 compounds). Of the compounds tested, shinjulactone K (1), ailanthone (2), shinjudilactone (3), and dehydrobruceantin (4) were the most potent. Although the activities were very low (0-19%), the resulting data provided a picture of structure-activity relationships. PMID- 9332007 TI - Synthesis of Tn and sialyl Tn antigen-lipid A analog conjugates for synthetic vaccines. AB - Conjugates of Tn and sialyl Tn antigen with N-teradecanoyl L-seryl-beta-alanine containing D-glucosamine derivatives structurally related to lipid A as an immunoadjuvant were synthesized for the development of totally synthetic vaccines against cancers or HIV. PMID- 9332006 TI - Preparation and characterization of polylactic acid microspheres containing bovine insulin by a w/o/w emulsion solvent evaporation method. AB - The objective of this study was to produce polylactic acid (PLA) microspheres containing bovine insulin as a sparingly water soluble model drug using a water in-oil-in-water (w/o/w) emulsion solvent evaporation method. The preparative conditions were optimized. Employment of smaller internal aqueous phase volume (50 microliters or 100 microliters) in the manufacturing process, resulted in the high loading efficiency (over 95% of theoretical insulin loading efficiency). The addition of 10% (w/v) NaCl to the external aqueous phase (0.5% polyvinyl alcohol solution) reduced loading efficiency compared to the case where no NaCl was added to the external phase. The mean volume diameter for prepared PLA microspheres was in the region of 15-25 microns in all cases. PLA microspheres containing 5% and 10% insulin theoretically exhibited burst release in the initial stage. After a three week dissolution test, the surface of the microspheres became more porous due to the degradable characteristics of PLA polymer itself. Nevertheless, about 80% of the insulin still remained undegraded in PLA microspheres. Finally, insulin-loaded PLA microspheres (corresponding to 4 I.U. insulin) were administered to normal rats subcutaneously, and the pharmacological effect (a decrease in serum glucose level) was demonstrated. PMID- 9332008 TI - Bovine serum albumin (BSA) as a reagent against non-specific immunogold labeling on LR-White and epoxy resin. AB - The purpose of this study was to examine how different incubation times with different concentrations of bovine serum albumin (BSA) affect the amount of non specific immunogold labeling on epoxy sections and LR-White sections. Immunogold labeling was performed on epoxy sections and LR-White sections of renal tissue with IgG-deposits and fibrin clots, and the antibodies used were anti-IgG and anti-fibrinogen, respectively. The sections were incubated with different concentrations of BSA prior to application of primary antibodies, and the length of this pre-incubation step varied between 0 and 4 h. During the incubation with primary antibodies, BSA was added in the same concentration as in the pre incubation step. The results showed that the non-specific labeling on the resin decreased significantly when the concentration of BSA or the length of the preincubation step was increased. The non-specific labeling was usually higher on the epoxy resin than on the LR-White resin when using the same conditions with respect to BSA. But, when the preincubation step with BSA lasted 4 h, the non specific labeling was somewhat lower on epoxy resin than on the acrylic LR-White resin, without respect to the concentration of BSA. The specific labeling for both fibrinogen and IgG decreased slightly when the concentration of BSA and incubation time increased, probably due to the steric hindrance performed by BSA molecules on the section. Blocking procedures with at least 1 h incubation time for the blocking step with at least 5% BSA are recommended for both epoxy and LR White sections. PMID- 9332009 TI - Systematic investigations of the contrast results of histochemical stainings of neurons and glial cells in the human brain by means of image analysis. AB - The investigation of neurohistological specimens by image analysis has become an important tool in morphological neuroscience. The problems which arise during the processing of these images are non-trivial, especially if a pattern recognition of cells in the imaged tissue is intended. One of the major problems faced concerns the segmentation of structures of interest, whether cells or other histologic structures. The segmentation problem is often the result of an inappropriate staining procedure. For serious image analysis to be performed, the material under investigation must be optimally prepared. Spatially complex patterns, e.g. fuzzy-like neighbouring neurons, are easy to recognize for humans. But the integrative and associative performance of current artificial neuronal network schemes is too low to achieve the same recognition quality as humans do. Therefore, a general analysis of staining characteristics was performed, especially with respect to those stains which are relevant to object segmentation. Although most image analytical investigations of tissues are based on stained samples, a study of this type has not been previously conducted. Of the stains and procedures evaluated, the gallocyanin chrome alum combination staining provided the best stain contrast. Furthermore, this staining method shows sufficient constancy within different parts of the human brain. Even the fine nuclear textures are differentiable and can be used for further pattern recognition procedures. PMID- 9332010 TI - Cerium-based ultracytochemical localization of aspartate transcarbamylase activity in the cell membrane complex of Saccharomyces cerevisiae. AB - Aspartate transcarbamylase (ATCase) activity was localized ultracytochemically in the yeast Saccharomyces cerevisiae by precipitation of its reaction product orthophosphate as cerium phosphate. We prefixed yeast cells with ice-cold 1% glutaraldehyde for 30 min which preserved 80% of ATCase activity. Cells were washed and incubated with ATCase substrates (aspartate, carbamyl phosphate) plus cerium chloride, and postfixed by osmium tetroxide. In cells from exponential batch cultures, deposits of cerium phosphate delineated simultaneously or alternatively membranes of the secretory pathway: nuclear envelope, endoplasmic reticulum, Golgi complex and the plasmalemma; mitochondrial membranes and intramitochondrial fibrous component were labelled as well. Deposits of cerium phosphate were never observed in the nucleoplasm. Cells incubated in the absence of cerium or ATCase substrates and mutant S. cerevisiae cells lacking ATCase activity served as controls. Small round electron-dense condensates were found to be randomly distributed within some cells, both in control and experimental runs, in the nucleoplasm, cytoplasm and mitochondrial matrix and represented undefined osmicated endogenous compounds. Our results suggest that the synthesis of pyrimidine precursors occurs in membranes, where compounds such as UDP-glucose and CDP-diglycerides are needed for membrane and/or yeast cell wall synthesis. The possible contribution of ATCase activity found in the nuclear envelope to nucleic acid synthesis remains to be clarified. PMID- 9332011 TI - Female space use is the best predictor of monogamy in mammals. AB - Monogamy is typically considered to have evolved either because biparental care is important for offspring survival, or because males are unable to monopolize more than one female due to females being too dispersed. Here, in the first phylogenetic analysis of the evolution of monogamy in mammals, we show that neither of these explanations is consistent with the distribution of monogamy across mammal species. Monogamy evolved significantly more often in the absence of paternal care than in its presence. Furthermore, monogamy does not normally occur in species where female ranges are large. Rather, the most common feature of mammalian monogamy is that it evolved where females were solitary and occupied small, exclusive ranges, enabling males to monopolize them. PMID- 9332012 TI - Heritability of pre-adult viability differences can explain apparent heritability of sperm displacement ability in Drosophila melanogaster. AB - Sperm displacement has been the subject of a large number of evolutionary studies because of its effects on relative male reproductive success. To understand better the evolutionary role of variation in sperm displacement ability (SDA), an obvious aim is to measure its heritability. In this paper, we show that a standard method used to measure the heritability of SDA can be misleading. First, we show that using conventional methods (based on counts of adult offspring of multiply mated females), SDA appears to be heritable. However, an examination of potentially confounding variables strongly suggests that this result is misleading, and that the heritable component is more likely to be pre-adult viability. Consequently, it is likely that there is little measurable heritable genetic variation for SDA in D. melanogaster. We conclude that, although conventional methods of measuring sperm displacement will usually be adequate for phenotypic measurements, greater care must be taken when measuring genetic variances. PMID- 9332013 TI - Adaptation to the fitness costs of antibiotic resistance in Escherichia coli. AB - Policies aimed at alleviating the growing problem of drug-resistant pathogens by restricting antimicrobial usage implicitly assume that resistance reduces the Darwinian fitness of pathogens in the absence of drugs. While fitness costs have been demonstrated for bacteria and viruses resistant to some chemotherapeutic agents, these costs are anticipated to decline during subsequent evolution. This has recently been observed in pathogens as diverse as HIV and Escherichia coli. Here we present evidence that these gentic adaptations to the costs of resistance can virtually preclude resistant lineages from reverting to sensitivity. We show that second site mutations which compensate for the substantial (14 and 18% per generation) fitness costs of streptomycin resistant (rpsL) mutations in E. coli create a genetic background in which streptomycin sensitive, rpsL+ alleles have a 4-30% per generation selective disadvantage relative to adapted, resistant strains. We also present evidence that similar compensatory mutations have been fixed in long-term streptomycin-resistant laboratory strains of E. coli and may account for the persistence of rpsL streptomycin resistance in populations maintained for more than 10,000 generations in the absence of the antibiotic. We discuss the public health implications of these and other experimental results that question whether the more prudent use of antimicrobial chemotherapy will lead to declines in the incidence of drug-resistant pathogenic microbes. PMID- 9332014 TI - Pigmented epithelium induces complete retinal reconstitution from dispersed embryonic chick retinae in reaggregation culture. AB - Reaggregation of dispersed retinal cells of the chick embryo leads to histotypic retinospheroids in which the laminar organization remains incomplete: photoreceptors form rosettes which are surrounded by constituents of the other retinal layers. Here, for the first time, a complete arrangement of layers is achieved in cellular spheres (stratoids), provided that fully dispersed retinal cells are younger than embryonic day E6, and are reaggregated in the presence of a monolayer of retinal pigmented epithelium (RPE). A remarkable mechanism of stratoid formation from 1 to 15 days in vitro is revealed by the establishment of a radial Muller glia scaffold and of photoreceptors. During the first two days of reaggregation on RPE, rosettes are still observed. At this stage immunostaining with vimentin and F11 antibodies for radial Muller glia reveal a disorganized pattern. Subsequently, radial glia processes organize into long parallel fibre bundles which are arranged like spokes to stabilize the surface and centre of the stratoid. The opsin-specific antibody CERN 901 detects photoreceptors as they gradually build up an outer nuclear layer at the surface. These findings assign to the RPE a decisive role for the genesis and regeneration of a vertebrate retina. PMID- 9332015 TI - Visual and socio-cognitive information processing in primate brain evolution. AB - Social group size has been shown to correlate with neocortex size in primates. Here we use comparative analyses to show that social group size is independently correlated with the size of non-V1 neocortical areas, but not with other more proximate components of the visual system or with brain systems associated with emotional cueing (e.g. the amygdala). We argue that visual brain components serve as a social information 'input device' for socio-visual stimuli such as facial expressions, bodily gestures and visual status markers, while the non-visual neocortex serves as a 'processing device' whereby these social cues are encoded, interpreted and associated with stored information. However, the second appears to have greater overall importance because the size of the V1 visual area appears to reach an asymptotic size beyond which visual acuity and pattern recognition may not improve significantly. This is especially true of the great ape clade (including humans), that is known to use more sophisticated social cognitive strategies. PMID- 9332016 TI - Global climate change and phenotypic variation among red deer cohorts. AB - The variability of two fitness-related phenotypic traits (body weight and a mandibular skeletal ratio) was analysed among cohorts and age-classes of red deer in Norway. Phenotypic variation among cohorts was pronounced for calves, yearlings and reproductively mature adults. Fluctuations in cohort-specific mean body weights and skeletal ratios of adults correlated with global climatic variation in winter conditions influenced by the North Atlantic Oscillation while cohorts were in utero. Red deer born following warm winters were smaller than those born after cold winters, and this inter-cohort variability persisted into adulthood. Phenotypic variation among cohorts of red deer influenced by climate change may pose consequences for fitness of cohorts since body size and condition contribute to reproductive success and survival in male and female red deer. In particular, the recent trend of increasingly warm winters in northern Europe and Scandinavia may lead to reduced body size and fecundity of red deer, and perhaps other ungulates, in those areas. PMID- 9332017 TI - Jaw muscle development as evidence for embryonic repatterning in direct developing frogs. AB - The Puerto Rican direct-developing frog Eleutherodactylus coqui (Leptodactylidae) displays a novel mode of jaw muscle development for anuran amphibians. Unlike metamorphosing species, several larval-specific features never form in E. coqui; embryonic muscle primordia initially assume an abbreviated, mid-metamorphic configuration that is soon remodelled to form the adult morphology before hatching. Also lacking are both the distinct population of larval myofibres and the conspicuous, larval-to-adult myofibre turnover that are characteristic of muscle development in metamorphosing species. These modifications are part of a comprehensive alteration in embryonic cranial patterning that has accompanied life history evolution in this highly speciose lineage. Embryonic 'repatterning' in Eleutherodactylus may reflect underlying developmental mechanisms that mediate the integrated evolution of complex structures. Such mechanisms may also facilitate, in organisms with a primitively complex life cycle, the evolutionary dissociation of embryonic, larval, and adult features. PMID- 9332018 TI - Detecting natural changes of cone-excitation ratios in simple and complex coloured images. AB - Ratios of excitations in each cone-photoreceptor class produced by light reflected from pairs of surfaces in a scene are almost invariant under natural illuminant changes. The stability of these spatially defined ratios may explain the remarkable ability of human observers to efficiently discriminate illuminant changes from changes in surface reflectances. Spatial cone-excitation ratios are not, however, exactly invariant. This study is concerned with observers' sensitivity to these invariance violations. Simulations of Mondrian paintings with either 49 or two natural surfaces under Planckian illuminants were presented as images on a computer-controlled display in a two-interval experimental design: in one interval, the surfaces underwent an illuminant change; in the other interval, the surfaces underwent the same change but the images were then corrected so that, for each cone class, ratios of excitations were preserved exactly. Although the intervals with corrected images corresponded individually to highly improbable natural events, observers systematically misidentified them as containing the illuminant changes, the probability of error increasing as the violation of invariance in the other interval increased. For the range of illuminants and surfaces tested, sensitivity to violations of invariance was found to depend on cone class: it was greatest for long-wavelength-sensitive cones and least for short-wavelength-sensitive cones. Spatial cone-excitation ratios, or some closely related quantities, seem to be the cues preferred by observers for making inferences about surface illuminant changes. PMID- 9332019 TI - Characterization of oligosaccharide composition and structure by quadrupole ion trap mass spectrometry. AB - The use of electrospray ionization-quadrupole ion trap mass spectrometry for the characterization of linear oligosaccharides and N-linked protein oligosaccharide mixtures is described. Tandem mass spectrometry (MS/MS) experiments with orders higher than two offer a number of ways to enhance MS/MS spectra and to derive information not present in MS and MS2 spectra. Three such methods are presented in this paper. (a) Collisional activation of permethylated oligosaccharide molecular ions (MS2) as illustrated by maltoheptaose, produces abundant fragments from glycosidic bond cleavages which indicate composition and sequence, and weak cross-ring cleavage products which denote specific linkages within the oligosaccharide. Through the trapping and further dissociation of these fragments (MSn), cross-ring cleavage products can be confirmed and their relative abundances increased to facilitate interpretation. (b) The mechanisms of formation of two isobaric ions or ions isobaric with another ion's isotope peaks, such as those present in the MS2 spectrum of the ribonuclease B oligosaccharide GlcNAc2-Man5 can be independently established by separate MS3 experiments. (c) Ions in the MS2 spectrum, specific for individual components of an isobaric mixture, can be isolated and characterized by further stages of fragmentation. This is illustrated by two isobaric oligosaccharides from chicken ovalbumin of the composition HexNAc5Hex5. These findings indicate the utility of ion trap mass spectrometry towards the facile determination of oligosaccharide composition, sequence, branching and linkage, providing a wealth of structural information not obtainable by other individual methods of carbohydrate mass spectrometric analysis. PMID- 9332021 TI - Discrimination of isomeric oligosaccharides and sequencing of unknowns by post source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - The PSD spectra of some isomeric oligosaccharides are investigated in order to study the fragmentation mechanisms. Results show that intense peaks are formed from one or more glycosidic linkage cleavages, while ring fragmentation produces very weak signals. Two analytical applications of post source decay (PSD) to the structural analysis of oligosaccharides are also illustrated: the discrimination of isomeric structures and the identification of a real unknown by combining PSD data with the linkage analysis results. PMID- 9332022 TI - The biosynthesis and metabolism of the aspartate derived amino acids in higher plants. AB - The essential amino acids lysine, threonine, methionine and isoleucine are synthesised in higher plants via a common pathway starting with aspartate. The regulation of the pathway is discussed in detail, and the properties of the key enzymes described. Recent data obtained from studies of regulation at the gene level and information derived from mutant and transgenic plants are also discussed. The herbicide target enzyme acetohydroxyacid synthase involved in the synthesis of the branched chain amino acids is reviewed. PMID- 9332023 TI - Endogenous protein phosphorylation and protein kinase activity in winged bean. AB - In winged bean (Psophocarpus tetragonolobus) protein kinases (E.C. 2.7.1.37) were found in all tissues studied. There was a significant increase in kinase activity during seed development, with a concomitant enhancement in the phosphorylation of a number of polypeptides; this was reversed in germinating seed cotyledons. Protein phosphorylation was apparently correlated with the increase in the protein content of the developing seed and the growing axis. At least three distinct autophosphorylating proteins could be distinguished in the developing seeds after SDS-PAGE, indicating the presence of different types of protein kinases in winged bean. PMID- 9332025 TI - Binding nature and denaturation of protein during interaction with galloylglucose. AB - Analysis of insoluble complexes between tetragalloylglucose and proteins following a series of successive washes with buffer indicated (1) heterogeneity of binding between galloylglucose and protein and (2) irreversible denaturation of protein during interaction with galloylglucose. Relatively large amounts of tetragalloylglucose were removed by initial washes, indicating weak, low affinity binding, whereas smaller amounts removed by subsequent washes suggest bonds with a higher affinity. Although the maximum number of bindings sites, calculated per 10,000 M(r) of protein, was similar for BSA, myoglobin and lysozyme, the proportion of these sites that appeared to have high affinity, varied from 8 to 29%. The low proportion of strongly binding sites in lysozyme explains its relatively low tannin-complexing ability. Solubility decrease in protein during successive washing and decrease in the beta-glucosidase activity indicate that irreversible denaturation of protein occurs, which progresses with an increase in the incubation time with galloylglucose and galloylglucose/protein molar ratio in the mixture. Relative affinity of galloylglucose is directly related to the ability to cause irreversible denaturation. PMID- 9332024 TI - Purification and characterisation of a protein kinase from winged bean. AB - A soluble, cytoplasmic protein kinase was purified from the developing seeds of winged bean (Psophocarpus tetragonolobus) following conventional methods of protein purification including anion-exchange chromatography, gel-filtration and Blue Sepharose chromatography. The purified enzyme consists of a single polypeptide of M(r) 45,000 as determined by SDS-PAGE and gel-filtration chromatography on Sephacryl S-200. The pH optimum of the protein kinase activity was 7.0, while the optimum concentration of Mg2+ was 5 mM. The enzyme utilised casein as an exogenous phosphate acceptor. The conventional modulators of protein kinases, including the cyclic nucleotides, Ca2+ and calmodulin, did not stimulate the purified enzyme. Heparin and spermine, too, had no effect on its activity. Phosphoamino acid analysis revealed that the enzyme transferred the gamma phosphate of ATP only to serine residues of casein. All these characteristics, taken together, classifies the purified protein kinase as a member of the casein kinase I group of enzymes. PMID- 9332026 TI - Cytotoxic flavonoids from Vitex agnus-castus. AB - Four new flavonoids, luteolin 6-C-(4"-methyl-6"-O-trans-caffeoylglucoside), luteolin 6-C-(6"-O-trans-caffeoylglucoside), luteolin 6-C-(2"-O-trans caffeoylglucoside), and luteolin 7-O-(6"-p-benzoylglucoside), together with four known ones 5, 4'-dihydroxy-3,6,7,3'-tetramethoxyflavone, luteolin, artemetin and isorhamnetin, were isolated from the root bark of Vitex agnus-castus. The structures were elucidated by spectroscopic means. PMID- 9332027 TI - [47th general meeting of the Japanese Society of Allergology. Tokyo, Japan. October 6-8, 1997. Abstracts]. PMID- 9332028 TI - [New options in the management of HIV-infection]. PMID- 9332029 TI - [Antiresorptive therapy for bone]. PMID- 9332030 TI - [Immunizations--a survey for the medical practice]. PMID- 9332031 TI - [50th annual meeting of the Japanese Association for Thoracic Surgery. October 1 3, 1997. Tokyo, Japan. Abstracts]. PMID- 9332032 TI - [The 70th congress of the Japanese Biochemical Society. Kanazawa City, Japan. September 22-25, 1997]. PMID- 9332033 TI - Amalgam arouses hot feelings. PMID- 9332034 TI - A model to meet the strategic information needs of dentistry in England. PMID- 9332035 TI - Agreement between practitioners concerning removal of asymptomatic third molars. AB - OBJECTIVE: To investigate and compare agreement within two groups of dental practitioners, family dentists and oral surgeons, in their decisions regarding removal of asymptomatic mandibular third molars. SUBJECTS: Ten oral surgeons and 18 family dentists from South Wales with experience ranging from 5 to 28 years. METHODOLOGY: Participants were presented with periapical radiographs of 36 asymptomatic, mandibular third molars and were informed of the age and gender of the patients and the degree of eruption of the third molars. Participants were asked to indicate whether they thought that the third molar should be removed or not. The degree of agreement between participants was measured by kappa indices for multiple raters. RESULTS: The kappa indices were 0.14 for the oral surgeons and 0.09 for the family dentists, indicating poor agreement beyond chance. Although in most cases the participants decided not to remove the third molar, they did so inconsistently, that is, they did not make this decision on the same cases. There were also differences in the inclination of the participants to suggest removal of the 36 third molars. CONCLUSION: Poor inter-observer agreement suggested that treatment decisions regarding asymptomatic third molars are based more on subjective beliefs and habitual practices than on rational decision making. PMID- 9332036 TI - Do item weights matter? An assessment using the oral health impact profile. AB - OBJECTIVE: To determine whether or not item weights contribute to the performance of the Oral Health Impact Profile (OHIP), a comprehensive measure of the functional, social and psychological outcomes of oral disorders. DESIGN: Data were obtained as part of an oral health survey of older adults living in Ontario, Canada. Subjects completed a personal interview, clinical examination and a self complete version of the OHIP. OHIP scores were calculated in three ways: a simple count method, an additive method and a method incorporating item weights derived from the Thurstone paired comparison technique. These scores were calculated for the full 49-item version of the measure and for a short form consisting of 14 selected items. The discriminant, concurrent and predictive validity of these scores for the two versions of the measure were ascertained. PARTICIPANTS: Complete data were obtained for 522 subjects. Just over half were female (56 per cent) and their mean age was 66 years. RESULTS: The OHIP discriminated between groups based on dental status (dentate/edentulous), presence of dry mouth (yes/no) and, for the dentate, according to the number of remaining teeth (less than 20/20 or more) irrespective of scoring method or the version of the questionnaire used. All scores showed significant associations with self-rated oral health, self-perceived need for dental care and dissatisfaction with oral health status. There was evidence to suggest that weighted scores were better at discriminating between groups than the simple count method but no better than the additive method. Similar findings emerged with respect to the ability of the scores to predict prosthodontic, surgical and restorative treatment needs. CONCLUSIONS: Although the data suggested that item weights did improve the performance of the OHIP, the fact that simple scoring methods were as good as more sophisticated ones might mean that the OHIP could be used in contexts, such as patient assessment for clinical care, where the calculation of weighted scores was not feasible. PMID- 9332037 TI - An interim determination of health gain from oral cancer and precancer screening: 1. Obtaining health state utilities. AB - OBJECTIVE: To obtain preliminary health state utility values for oral precancer, and stage 1 and stage 2 (or greater) oral cancer, for use in a study involving quality of life measurement. DESIGN: Members of the general public were personally interviewed and asked to complete a standard gamble questionnaire in which each individual had to choose between having oral cancer or precancer (a description of each stage and its treatment was given), and being perfectly healthy but with a probability (p) from 0 to 1, of immediate death. There were three questions, the first related to a state of oral precancer, the second to a small cancer (< 2 cm) and the third to a large oral cancer (> 2 cm). The utility of each state was equal to the probability that the subject would opt for having the disease (1-p). The pooled responses of all subjects were analysed using a Wilcoxon matched-pairs signed-ranks test. SETTING: The premises of a commercial company. PARTICIPANTS: A convenience, quota sample of 100 employees aged 40 years or over. RESULTS: The mean utility values obtained were: oral precancer = 0.92; small oral cancer = 0.88; large oral cancer = 0.68. There was a significant difference between all three states (P < 0.05). CONCLUSIONS: The study relied on a convenience sample to elicit the public's perceptions of different oral cancer states. Nevertheless, despite this limitation, it was considered that the utility values derived were suitable for incorporation in measurements of quality adjusted life years. PMID- 9332038 TI - Risk factors associated with tooth wear in teenagers: a case control study. AB - OBJECTIVE: To measure the association of various aetiological risk factors with tooth wear in 15-year-old school children. BASIC RESEARCH DESIGN: A case control study conducted one year after a prevalence study (Milosevic et al., 1994). The 80 children, identified from the prevalence study with palatally and/or occlusally exposed dentine, acted as cases. The control group were systematically selected from every tenth name on year registers. The only matching criteria were age, gender and school. All children answered a questionnaire detailing the risk factors and the tooth wear was further scored by one examiner (AM). SETTING: Ten randomly selected schools in Liverpool. PARTICIPANTS: 102 children participated out of a possible 160, an overall response rate of 64 per cent. Of the 54 controls, six had developed tooth wear into dentine during the 12-month interval following the prevalence study and were therefore excluded from the analysis. RESULTS: The odds ratio on 37 matched pairs was 1.7 (95% CI 0.72, 3.87) for grinding and clenching teeth and 2.6 (95% CI 0.91, 7.45) for carbonated drink consumption. Eating pickled food other than pickled onions was significantly more frequent in cases than in controls. Other potential aetiological factors which were not significantly associated with tooth wear in this group of children included stomach upsets, frequency of tooth brushing, weight/body shape and drinking fruit juice or using ketchup/sauces. Conditional logistic regression analysis of the paired data showed that carbonated beverage consumption was on the borderline of significance (P = 0.055) as a predictor for tooth wear in teenagers either on its own or allowing for the effect of tooth grinding. CONCLUSIONS: The frequent consumption of carbonated beverages is probably related to tooth wear. Future case control studies should seek to incorporate around 100 to 160 cases. PMID- 9332039 TI - The prevalence, causes and cosmetic importance of dental fluorosis in the United Kingdom: a review. AB - OBJECTIVES: To determine the prevalence of fluorosis and other developmental defects of enamel (DDE) in the United Kingdom over the last 40 years. To examine the risk factors that may have influenced the prevalence of fluorosis. DESIGN: All relevant publications between 1956-1995 on the prevalences of DDE and fluorosis in the UK were critically reviewed in addition to publications from other countries where these added substantial information about the risk factors involved in the latter. RESULTS: The likely overall prevalence of DDE in the UK is approximately 40 per cent of which diffuse opacities account for about half. In areas benefiting from water fluoridation the overall prevalence of DDE is around 70 per cent with diffuse opacities accounting for two thirds. No substantial evidence of increases in either DDE or fluorosis could be identified. Risk factors for fluorosis include inappropriate use of fluoride supplements and fluoride toothpaste during early childhood and life-time residence in an optimally fluoridated area. CONCLUSIONS: Fluorosis does not appear to be of public health concern in the UK at present. However, the inadequacies of the data and paucity of in-depth studies highlight the need for further co-ordinated research using agreed methods of monitoring prevalence, multivariate analysis to identify risk factors and a societal norm to discover its cosmetic impact. PMID- 9332040 TI - Jarman underprivileged area scores, tooth decay and the effect of water fluoridation. AB - OBJECTIVE: To examine the association between dental caries and Jarman underprivileged area scores at regional, district and electoral ward level and explore any possible relationship with water fluoridation. DESIGN: An ecological study using the English results from the NHS dental surveys on 5-year-old children in 1991/2 and 1993/4, and the survey of 12-year-old children in 1992/3. Jarman underprivileged area scores were used from the 1991 census. SETTING: The study used former English health authority regions and districts. The electoral wards were in non-fluoridated Salford and Trafford and Liverpool, and fluoridated Newcastle and North Tyneside. SUBJECTS: The random sample of 5-year-old children examined in 1991/2 and 1993/4, and 12-year-old children in 1992/3, in studies coordinated by the British Association for the Study of Community Dentistry. OUTCOME MEASURES: Correlations between regional, district and electoral ward mean dmft/DMFT and Jarman underprivileged area scores. RESULTS: Significant correlations were demonstrated at a regional level in 5-year-old children in 1991/2, district level in 5-year-old children in 1991/2 and 1993/4 and at electoral ward level in both age groups in 1992/3 and 1993/4. Correlation coefficients varied between r = 0.88 to r = 0.46. Multiple linear regression at electoral ward level showed significant interactions between Jarman scores and water fluoridation. There was an average 44 per cent and 43 per cent reduction in caries in fluoridated electoral wards in 5- and 12-year-old children respectively. In deprived electoral wards, with a Jarman score of 40, the reduction in caries increased to 54 per cent and 56 per cent for 5- and 12-year old children respectively. CONCLUSIONS: Dental caries was associated with Jarman underprivileged area scores. The differential efficacy of fluoridation to deprived areas was demonstrated at electoral ward level. Water fluoridation was confirmed as an evidence based intervention which has halved the amount of tooth decay in 5- and 12-year-old children. PMID- 9332041 TI - On the assessment of dental health care attitudes in 1986 and 1995, using the dental attitude questionnaire. AB - OBJECTIVE: To re-establish and update the empirical data obtained in 1986 with the Dental Attitude Questionnaire. DESIGN: In 1995 this questionnaire, presented earlier by Hoogstraten and Broers (1986), was completed in a similar setting using similar subjects as in 1985, to make a comparison between 1986 and 1995 possible. SUBJECTS: 375 persons, all first grade psychology students who participated for additional course credit. Mean age was 21.7 years, 65 per cent were female. RESULTS: Data show a change in oral health care attitudes and a change in the internal consistency of the DAQ subscales, making the present version of the questionnaire inadequate for measuring present oral health care attitudes. CONCLUSION: This study has shown once more the importance of conducting replication studies after relatively long periods of time in order to update the psychometric characteristics of questionnaires. PMID- 9332042 TI - Dentists' decisions on caries risk and preventive treatment by dental state among 15-year-old adolescents. AB - OBJECTIVE: To evaluate dentists' decisions on caries risk and preventive treatment in relation to their clinical findings among 15-year-old children. DESIGN: Two groups of children were selected on the basis of subjects' number of decayed teeth. Data were also used from personal oral health records. PARTICIPANTS: 15-year-old children (n = 132) clinically checked and treated in public dental clinics in Helsinki. The high-risk group (n = 100) had the greatest numbers of decayed teeth (DT + dt), the low-risk group (n = 32) was cavity-free. OUTCOME MEASURES: Dental state, treatment decision, preventive and operative treatment recorded by visit, were used for assessment of caries risk and individual needs for caries prevention, and for evaluation of diversity and intensity of preventive treatment given to each patient. RESULTS: Dentists had judged only 17 per cent of the high-risk patients (mean DT + dt 6.5; range 4-16) as being at high risk of caries. Preventive measures were given only on one out of three visits. One out of two preventive measures was application of fluoride varnish and only one out of ten was dietary counselling. Patients with more decayed surfaces had been given more diversified (P < 0.01) and more intensive (P < 0.005) preventive treatment than had those with none or only a few caries lesions. Despite this, only 4 per cent of the patients with DS = 8-27 had been given four different preventive measures, and one patient out of seven had been left without any caries-preventive intervention. CONCLUSION: To meet individual needs of high caries-risk patients, the variety and intensity of preventive measures directed towards them need to be further improved. PMID- 9332043 TI - Prevalence of dental caries in Omani 6-year-old children. AB - OBJECTIVE: To determine the prevalence of dental caries in Omani 6-year-old children. DESIGN: Clinical caries cross-sectional survey, conducted by 16 trained and calibrated dentists. SETTING: Omani primary schools in December 1994. SUBJECTS: 3,114 subjects, randomly selected to achieve an overall 6.6 per cent sample of Omani 6-year-old children. OUTCOME MEASURES: Caries diagnosis based solely on clinical examination in accordance with criteria of British Association for the Study of Community Dentistry. RESULTS: Only 484 subjects (15.5 per cent) were caries-free; regional variations ranging from 4.4 per cent to 31 per cent. Overall, the national dmft averaged 4.61; the majority of caries experienced being in the form of untreated decay, with occlusal surfaces of first primary molars being the most commonly involved site. CONCLUSIONS: Compared with results from a survey of Omani 12-year-old children in 1993, a much smaller proportion of this 6-year-old sample were caries free, emphasising the need for continuance of existing, and the development of further, preventive programmes. PMID- 9332044 TI - An evaluation of a measure of subjective oral health status in the UK. AB - OBJECTIVE: To evaluate the performance in the United Kingdom of an instrument to measure the subjective impact of oral conditions; that is, the functional restrictions and symptoms, as well as the disability and disadvantage experienced as a result of oral problems. BASIC RESEARCH DESIGN: The instrument tested was the Subjective Oral Health Status Indicators (SOHSI) that consists of a battery of eight subjective indicators and which has been developed and tested in Canada. A questionnaire containing these indicators was administered by post to two random samples of two hundred and fifty residents aged 60-65 years, one from an affluent and one from a deprived electoral ward in Liverpool. MAIN OUTCOME MEASURES: Reliability was assessed as test-retest, and internal consistency using Cronbach's alpha. Construct and concurrent validity were assessed by determining the association between the subjective indicators and dentate status, reported satisfaction and overall subjective assessment of oral health. The constituent concepts of the theoretical model on which the instrument is based were tested by correlating the subjective indicators and the self-reported number of teeth. CONCLUSIONS: The instrument was found to be reliable both in terms of test-retest reliability and internal consistency. It also demonstrated satisfactory construct and concurrent validity. Correlations between self-reported number of teeth and the subjective indicators confirm the strength of the theoretical model on which the instrument was based and provide further evidence of the content validity of this composite measure. PMID- 9332045 TI - [Bacterial culture of chip tissue of enucleated prostates]. AB - To assess the prevalence of infection and colonization of the prostate by bacteria, chip tissue samples from 166 patients undergoing retropubic prostatectomy were submitted for bacterial tissue culture. In 28 patients with an indwelling catheter before surgery, E. coli, Klebsiella, Pseudomonas and Enterobacter were the commonest species encountered, the first the most common. In only 7 patients (20%) who didn't have an indwelling catheter before operation was the culture positive. We confirmed that the longer the time the catheter was indwelling before surgery, the greater the likelihood of positive cultures. However, postoperative outcome and morbidity were not related to culture results. We conclude that even though it is worth trying to sterilize the urine and prostate before prostatectomy, the effect on the postoperative outcome is minimal when proper antimicrobial therapy is given perioperatively. PMID- 9332046 TI - [Therapeutic and toxic theophylline levels in asthma attacks--is there a need for additional theophylline?]. AB - Although first-line therapy for bronchial asthma has changed over the past decade to anti-inflammatory medication such as inhaled corticosteroids and cromolyn with possible addition of beta-agonists, theophylline is still useful and therefore widely used. However, several studies have raised serious questions regarding its efficacy in acute asthmatic exacerbations. These studies, the narrow therapeutic range of the drug, the frequency of side effects and interactions with common drugs, and individual variation in clearance and metabolism, have prompted its reevaluation in the management of asthma. Therapeutic serum levels of theophylline are between 10 to 20 mcg/ml. Most adults achieve these concentrations with daily slow-release oral theophylline preparations, 200-400 mg (approximately 10 mg/Kg) twice a day. However, when such a patient presents to the emergency room (ER) in an asthmatic attack, immediate intravenous theophylline is often given, regardless of maintenance treatment. Since the rationale for this common therapeutic approach has been challenged, the current study was undertaken. Serum theophylline levels were measured in 23 consecutive asthmatics presenting to the ER in an acute attack. 15 (68%) had therapeutic levels (above 10 mcg/ml) and 2 had toxic levels (above 20 mcg/ml), prior to receiving the standard intravenous theophylline dose given for an attack. These data indicate that most patients with bronchial asthma on oral maintenance theophylline do not require additional intravenous theophylline when in an attack. It probably will not benefit them and may even induce serious theophylline toxicity. PMID- 9332047 TI - [Surgical approach to benign extradural lesions of the thoracic spine]. AB - A benign epidural lesion in the thoracic spine is rare, and usually the result of intervertebral disc herniation or infection. Not long ago patients were diagnosed late in the course of their disease and the surgical results of the standard laminectomy usually performed were grave. The development of newer imaging techniques (CT and MRI) has made diagnosis much easier, so diagnosis is often earlier, when neurological deficit is minimal. Newer neurosurgical techniques and approaches to the thoracic spoine have been developed to treat these lesions, which we describe. Clinical data on 16 patients operated from January 1996 to January 1997 are presented. PMID- 9332048 TI - [Terbinafine-induced cholestatic liver injury]. AB - A 43-year-old man presented with weakness, pruritus, skin rash and jaundice 2 weeks after treatment for onychomycosis with terbinafine (Lamisil) was started. Liver function tests showed combined hepatocellular and cholestatic injury. Ultrasound examination, computerized tomography and ERCP excluded extrahepatic obstruction. Serology was negative for HBV, HCV, HAV, CMV, and EBV. Liver biopsy was consistent with drug-induced cholestatic injury. Since the clinical picture did not improve when terbinafine was stopped, corticosteroids were started and resulted in complete clinical and laboratory recovery; liver function tests were normal 8 months after corticosteroids were discontinued. PMID- 9332049 TI - [Gastrointestinal angiodysplasia as a cause of hemorrhage in the digestive tract and treatment]. AB - Gastrointestinal angiodysplasia is a cause of gastrointestinal bleeding in the elderly, for which surgery has been the only treatment. Estrogen has been reported beneficial in some cases in the past decade. Recurrent bleeding due to angiodysplasia occurred from the small intestine in a 75-year-old woman, and from the right colon in a 91-year-old man. The diagnoses were made by angiography in the first case and colonoscopy and erythrocyte- scanning in the second. There was aortic stenosis in both, a combination which has been reported in other cases. Both patients improved with estrogen therapy. However, after temporary stabilization, gastrointestinal bleeding recurred in the second patient and he was successfully operated on. PMID- 9332050 TI - [Surgical repair of fractures of the clavicle in the adult]. AB - Clavicular fractures make up 45% of shoulder girdle fractures. The clavicle's susceptibility to injury is due to its subcutaneous position and its role as the bony connection between the thorax and the shoulder. In 95% of cases the mechanism of injury is a direct blow to the shoulder. These fractures are usually treated conservatively without surgery. But there are a few such fractures that require surgical repair in order to unite well. 9 patients were operated on for clavicular fractures during 1991-1995. The indications for surgical repair were lateral-third fracture, floating shoulder, neurovascular deficit or nonunion. The methods used were open reduction and fixation with either plate and screws, Kirchner wires, cerclage or a combination. All fractures united well, with no infections or new neurovascular deficits. Good range of shoulder motion and acceptable cosmetic results were achieved in all. 1 patient had functional limitation due to brachial neuritis caused by brachial damage at the time of injury. Indications for surgical repair and the methods used in these cases are similar to those described in the literature. The high rate of union and absence of complications support surgical repair for the few calvicular fractures that are not likely to unite properly. PMID- 9332051 TI - [Left atrial ball thrombus]. AB - An 80-year-old hypertensive woman with chronic atrial fibrillation was hospitalized because of recurrent syncope. Echocardiography revealed a large left atrial ball thrombus. Operative findings confirmed the echocardiographic diagnosis. PMID- 9332052 TI - [The enigma of Kaposi's sarcoma--the end of the matter?]. PMID- 9332053 TI - [Which scientific article is best qualified for publication?]. PMID- 9332054 TI - [Open heart surgery in dialysis patients]. PMID- 9332055 TI - [Taxanes: a breakthrough in cancer chemotherapy]. PMID- 9332056 TI - [Dissociative identity disorder in children and adolescents]. PMID- 9332057 TI - [Candida albicans vulvovaginitis--trends in care and implications]. PMID- 9332058 TI - [Narcolepsy]. PMID- 9332059 TI - [Laparoscopic surgery: the postoperative advantage versus the intraoperative risk, or are the sick too sick]. PMID- 9332060 TI - [Refusal of invasive obstetrical intervention and the patient's right law]. PMID- 9332061 TI - [Mangled extremity syndrome]. PMID- 9332063 TI - [Hospital restructuring and reorganization]. PMID- 9332062 TI - [New antipsychotic drugs]. PMID- 9332064 TI - [Penetrating gluteal injuries management--the need for management protocols]. AB - Gunshot wounds are the most common cause of penetrating injuries in the gluteal region (PIGR), while only 17.5% result from stabbing. Complications and even death have been described as resulting from stab injury in this area, which is thus a hazard that demands an algorithm for proper management. Recently, DiGiacomo et al. recommended that transpelvic bullet trajectory warrants surgery. Mercer et al., in a study of 81 patients with PIGR, recommended an algorithm based on anatomical gluteal zones and concluded that angiography is not mandatory in such injuries. We describe 9 cases of PIGR, 4 of which were from stab wound and 1 a gunshot wound, which bled from rami of the gluteal arteries. In 2 there was no apparent bleeding, and they were hemodynamically stable. Thus in this type of injury significant damage may be obscured. Our results suggest that angiography is important in the evaluation of such injuries an should be part of the protocol for management of PIGR. PMID- 9332065 TI - [Permanent pacemakers in the chest X-ray]. PMID- 9332066 TI - [A visit to Cairo Children's Hospital]. PMID- 9332067 TI - [Impaired physicians: the Israel Intervention Program]. AB - Diagnosis and treatment of mental disorders among physicians is a complicated problem. In western countries alcohol and drug abuse are the main mental problems of physicians, and programs of early treatment help them keep their practices without their harming their patients. We describe the activities of the Israel Committee for Psychiatric Examination of Physicians in 1989-93, when 115 physicians were examined. Addiction to alcohol or drugs was rare, but mental illness was detected in 50% of those examined. The results indicated that the committee acted therapeutically, dictating mental treatment and following up on it. Due to this policy, in most cases physicians were enabled and allowed to continue their practices. The results emphasize the need for occupational mental health facilities for physicians and other medical professionals. PMID- 9332068 TI - [Gynecologic problems of the lower genital tract in children and young adolescents]. AB - Hospital records of 46 girls under the age of 17 years, hospitalized for lower genital tract problems in 1986-95 were reviewed. The most common conditions were results of unintentional injuries (43.5%), imperforate hymen (28.2%) and infections (19.6%). The median age for unintentional injuries was significantly lower than for other conditions (7.0 vs 11.4; p < 0.001). Most injuries were external and occurred during outdoor activities. Mean volume of estimated bloody fluid drained in those with imperforate hymen was greater when the diagnosis was made after the age of 12 (783 vs 433; not significant). It has been suggested that hematocolpos and hematometra should be prevented, but the possible unfavorable sequelae have not been documented. The relative order of frequency of the various diagnostic groupings and the diagnoses of labial adhesions and imperforate hymen are specific for the age of the study group. PMID- 9332069 TI - [Radical retropubic prostatectomy]. AB - Radical prostatectomy may cure most patients in whom the malignant tumor has not invaded through the prostatic capsule. Advances in surgical technique and accumulation of experience have decreased the complication rate significantly. Long-term results of surgical treatment are now better than those of other forms of treatment; hence radical prostatectomy is now recommended for men with life expectancies longer than 10 years. Between 1988 and 1995, 164 men with clinical stages T1 or T2 adenocarcinoma were admitted for radical prostatectomy. Most were not offered a nerve-sparing procedure, so as to allow wider, more complete resection. Those who wanted preservation of sexual function underwent the nerve- preserving procedure. In 6 patients operation was discontinued when metastases to the mac lymph nodes were detected and in 1 when invasion of the pelvic wall was found, 157 underwent radical prostatectomy. Preoperative biopsy revealed a low grade lesion (Gleason 2-4) in 19.1%, intermediate grade (Gleason 5-6) in 61.8% and high-grade (Gleason 7-9) in 19.1%; however, pathologic grading revealed that only 7.0% had grade 2-4 tumor, 60.5% grade 5-6 and 32.5% grade 7-9. Pathologic staging revealed T2 tumor in 58%, T3 in 38.8% (including microscopic invasion of the capsule or seminal vesicles); microscopic lymph node metastases were found in 3.2%. Tumor invasion through the capsule was found in only 2 of 13 treated with neoadjuvant androgen blockade, compared with 40% in those who did not receive this treatment. There was no operative mortality and only 14.7% has complications. All had urinary incontinence immediately after operation, but regained continence after an average of 4-5 months, 24 were incontinent for more than 12 months, but most of them had only mild stress incontinence. Most patients were impotent after the procedure. There was tumor recurrence, diagnosed by rise in serum PSA, in 26 during an average followup of 26.4 months (range 3-93). Cure rate of prostatic cancer by radical prostatectomy may be increased by improved preoperative staging methods and better patient selection; long term follow up is required for determining cure rate. PMID- 9332071 TI - [Buschke-Ollendorf syndrome]. AB - Buschke-Ollendorf syndrome is a rare condition characterized by uneven sclerotic, osseous formations seen on X-ray (osteopoikilosis) and fibrous skin papules (dermatofibrosis lenticularis disseminata). We report an 82-year-old man with this syndrome. Awareness of the condition is important to avoid misdiagnosis and hazardous management designed for other disorders, such as prostatic metastases. PMID- 9332070 TI - [Hemato-oncology patients in acute respiratory failure in the ICU]. AB - Hemato-oncology patients needing mechanical ventilation for acute respiratory failure (ARF) have an extremely poor prognosis, with a mortality of more than 90%. Over an 18 month-period 17 such patients were admitted to our ICU. Diagnoses included leukemia (11 cases), lymphoma (1), and status post bone marrow transplantation for leukemia, lymphoma or breast cancer (5). Of 8 whose ARF was associated with septic complications due to neutropenia following chemotherapy, 6 survived. Of 9 who developed ARF due to toxic damage to vital organs following high-dose chemotherapy, 2 survived. Those who develop ARF during chemotherapy are expected to have an increase in granulocyte count within days, and have a surprisingly good prognosis. They should be admitted to the ICU and treated aggressively. Those who develop sepsis due their primary disease and whose general condition contraindicates chemotherapy, have an extremely grave prognosis and admission to the ICU may not be warranted. PMID- 9332072 TI - [Budd-Chiari syndrome]. AB - Budd Chiari syndrome is a rare disorder resulting from occlusion of hepatic venous drainage by hepatic vein thrombosis or by a membranous web in the inferior vena cava. In western countries the commonest causes are myeloproliferative disorders and hypercoagulable states. Presentation may be acute with rapid accumulation of ascites and hepatic failure, or subacute with symptoms developing over a few months. A chronic progressive form has also been described. On presentation there is usually abdominal pain, ascites, and hepatosplenomegaly; hepatic encephalopathy is found in about a third. Noninvasive, ultrasound-Doppler is recommended in diagnosis, and has a high correlation with hepatic venography. Liver biopsy is required for therapeutic decisions. Those with advanced hepatic failure or severe fibrosis on liver biopsy are referred for hepatic transplantation. When biopsy shows only hepatic congestion and inflammatory infiltrates, portosystemic shunting is recommended. We present a 61-year-old woman with ascites and hepatosplenomegaly that had developed over the courses of a few months. Budd-Chiari syndrome with chronic myelofibrosis and congenital protein C deficiency were diagnosed. Portosystemic shunt was performed but death from sepsis followed shortly. PMID- 9332073 TI - [Salt--hypertension and other possible adverse effects]. PMID- 9332075 TI - [Pain in infants]. PMID- 9332074 TI - [Colchicine as therapy for recurrent pericarditis]. PMID- 9332076 TI - [Large scale screening for Gaucher's disease in Israel--a position paper by the National Gaucher Committee of the Ministry of Health]. PMID- 9332077 TI - [The new law of patient privilege--issue of medical confidentiality]. PMID- 9332078 TI - [New generation of oral contraceptives and the cardiovascular effects]. PMID- 9332079 TI - [Specialty medicine--where? Hospital versus community]. PMID- 9332081 TI - [Ocular involvement in cancer therapy]. PMID- 9332080 TI - [Active specific immunotherapy of malignant melanoma]. PMID- 9332082 TI - [A diagnostic approach to soft tissue sarcomas]. PMID- 9332083 TI - [Multidrug resistance: its significance in hematological malignancies and its role in normal tissues and blood cells]. PMID- 9332084 TI - [Mechanical ventilation for acute respiratory failure in chronic obstructive pulmonary disease]. PMID- 9332085 TI - [Medical error--incidence and prevention]. PMID- 9332086 TI - [Circumcision and urinary tract infections]. PMID- 9332087 TI - [Thermal treatment of the painful joint--to heat or to cool]. PMID- 9332088 TI - [Trauma registry--the concept and its application in Israel]. PMID- 9332089 TI - [Abdominal pain, vomiting and systolic murmur in a 13-year-old girl]. PMID- 9332090 TI - [Hemorrhagic diathesis]. PMID- 9332091 TI - Consumer protection law and the pediatrician. PMID- 9332092 TI - Hemoglobin E-beta thalassemia in Uttar Pradesh. AB - OBJECTIVE: To evaluate the molecular make up of hemoglobin E-Beta thalassemia to facilitate diagnosis, genetic counseling and prenatal diagnosis in Uttar Pradesh. DESIGN: DNA analysis. SETTING: Referred hemolytic anemia cases to Genetics OPD of a tertiary care center. SUBJECTS: 21 families of HbE-thalassemia of which 19 were of UP origin. METHODS: The patient and obligate carriers in their families were evaluated at hematological, biochemical and molecular level. A total of 62 cases were evaluated which included the index cases and their family members. Red blood cell indices, osmotic fragility, hemoglobin electrophoresis, quantitation of fetal hemoglobin, HbA2/E, serum iron and total iron binding capacity estimation were carried out in all the blood samples. DNA analysis was done for HbE and beta thalassemia mutations. RESULTS: The commonest, IVSI-5 (G-->C) mutation (57%) was found along with HbE mutation. Only 23/26 cases belonged to the group of common beta-thal mutations as described in literature. CONCLUSION: Establishment of antenatal diagnostic services is necessary in those parts of India where both these mutations are commonly seen. PMID- 9332093 TI - Effect of storage of colostrum in various containers. AB - OBJECTIVE: To compare the effect of storage on expressed colostrum kept at room and refrigeration temperature in 2 different types of containers (steel and plastic) at different time intervals (0 hours to 7 hours). DESIGN: Prospective immunological study. SETTING: Maternity ward. METHODS: Colostrum was collected from 60 healthy lactating mothers and tested for total and differential cell counts and cell viability in plastic (polypropylene) and steel containers at 0 hours and 7 hours after storage at 28 degrees C and 4 degrees C. RESULTS: Colostrum stored for 7 hours in plastic containers had a significantly (p < 0.001) higher cell count and viability compared to that stored in steel containers both at 4 degrees C and 28 degrees C. The differential cell count did not vary with time, temperature or storage vessel. CONCLUSION: If required, colostrum should be stored at 4 degrees C in plastic (polypropylene) containers to maintain its protective quality. PMID- 9332094 TI - Neonatal nosocomial infection: profile and risk factors. AB - OBJECTIVE: To determine the incidence and risk factors for neonatal nosocomial infections. DESIGN: Cohort study. SETTING: Tertiary care Teaching Hospital. METHODS: Hospital born neonates transferred to the neonatal unit after birth and available in the unit 48 hours later comprised the cohort for the surveillance. Detailed maternal, intrapartum and neonatal variables were recorded. Risk factors for nosocomial infection were analyzed by both univariate and multiple logistic regression methods. RESULTS: One hundred and thirty-four neonates were enrolled in the cohort. The overall nosocomial infection rate was 16.8/1000 patient days. Device associated infection rate was 11.9/1000 device days. Multidrug resistant Klebsiella species was the commonest organism causing nosocomial septicemia and pneumonia followed by Pseudomonas aeruginosa. The risk factors detected to be significantly associated with infection on multiple logistic regression analyses were a birth weight < 1500 g (OR 3.3) and assisted ventilation > 72 h (OR 14.2). CONCLUSIONS: Very low birth weight (VLBW) neonates, especially those undergoing interventions such as mechanical ventilation are at the greatest risk for infection and death. Therefore, strict protocol for asepsis must be adhered to when handling these high risk infants. PMID- 9332096 TI - Use of carotene-rich foods to combat vitamin A deficiency in India--a multicentric study by the Nutrition Foundation of India. PMID- 9332095 TI - Anti-reflux therapy. PMID- 9332097 TI - Consumer Protection Act and medical profession. PMID- 9332098 TI - Apnea in neonates. PMID- 9332099 TI - Parental presence during anesthesia induction. PMID- 9332100 TI - Mycobacterial lymphadenitis. PMID- 9332101 TI - Status of iodine deficiency in selected blocks of Kangra District, Himachal Pradesh. PMID- 9332102 TI - Xeroderma pigmentosum. PMID- 9332103 TI - A 12-year-old boy with fever and blue ears. PMID- 9332104 TI - Adenocarcinoma colon. PMID- 9332105 TI - Monosomy 22 mosaicism. PMID- 9332106 TI - Early hemorrhagic disease of newborn. PMID- 9332107 TI - Transportation of sick neonates. PMID- 9332108 TI - Repeat process evaluation of pulse polio immunization. PMID- 9332109 TI - Neurodevelopmental outcome with early indomethacin. PMID- 9332110 TI - Multi-drug resistant Salmonella typhimurium. PMID- 9332111 TI - Serodiagnosis of tubercular infection. PMID- 9332112 TI - Intravenous magnesium sulfate in acute severe asthma not responding to conventional therapy. AB - OBJECTIVE: To evaluate the effectiveness of early administration of intravenous Magnesium sulfate (i.v. MgSO4) in children with acute severe asthma not responding to conventional therapy. DESIGN: Randomized double-blind, placebo controlled trial. SETTING: Pediatric emergency service of a large teaching hospital. SUBJECTS: 47 children aged between 1-12 years with acute severe asthma showing inadequate or poor response to 3 doses of nebulized salbutamol given at an interval of 20 min each. INTERVENTION: The MgSO4 group received 0.2 ml/kg of 50% MgSO4 as intravenous (i.v.) infusion over 35 minutes and the placebo group received normal saline infusion in the same dose and at the same rate. MgSO4 solution and normal saline were coded and dispensed in identical containers. Decoding was done at the completion of the study. All the patients received oxygen, nebulized salbutamol, i.v. aminophylline and corticosteroids. RESULTS: MgSO4 group showed early and significant improvement as compared to placebo group in PEFR and SaO2 at 30 min and 1, 2, 3 and 7 hours after stopping the infusion (p ranging from < 0.05 to < 0.01). The clinical asthma score also showed significant improvement in the MgSO4 group 1, 2, 3 and 11 hours after stopping the infusion (p < 0.01). CONCLUSION: Addition of MgSO4 to conventional therapy helps in achieving earlier improvement in clinical signs and symptoms of asthma and PEFR in patients not responding to conventional therapy alone. PMID- 9332113 TI - Sensitivity of neonatal tetanus surveillance system in India. AB - OBJECTIVE: To estimate the sensitivity of neonatal tetanus (NNT) surveillance in India. DESIGN: A comparison of two sets of data obtained from NNT mortality surveys and routine surveillance system. METHODS: NNT mortality surveys were undertaken in 1981, 1989 and 1992 using 30 cluster sampling technique. The data on reported incidence of NNT through routine surveillance system was taken from the published documents of Health Ministry and WHO. RESULTS: In 1981, the incidence of disease in a national survey was estimated to be 4 and 16.4 per 1000 live births in urban and rural areas, respectively. Follow up surveys in 1989 and 1992 estimated the overall incidence as 4 and 1.74 per 1000 live births, respectively. Comparing the reported and estimated by surveys, around 10% of NNT cases were reported. CONCLUSIONS: There is an urgent need to strengthen the routine surveillance system which at present grossly under-reports the NNT incidence in India. PMID- 9332114 TI - Prediction of birth weights from body weights of newborns. AB - OBJECTIVE: To evaluate the accuracy of prediction of birth weights from body weights of newborns till six days after birth. DESIGN: Prospective follow-up. SETTING: Four villages near Hyderabad. METHODS: Weights of 47 newborns were recorded daily from the day of birth for seven days. The birth weights were regressed on the weights of the babies taken on the 2nd day to the 7th day. Specificity and sensitivity of the predicted birth weights to arrive at the prevalence of low birth weight (LBW) were computed. RESULTS: The co-efficient of determination (R-square) for between the days measurements decreased from 95% on the second day to 86% on seventh day with an increase in the standard error of the estimate from 84 g to 154 g. Based on the "predicted birth weights", the prevalence of LBW in the community was arrived at and compared with the actual observation. The sensitivity and specificity of these regression equations was high and ranged from 0.95 to 0.85 and 0.96 to 0.93, respectively. CONCLUSIONS: In situations where the birth weight cannot be recorded, weight of the baby taken within the first week after birth may be reliably utilized to assess the "birth weight", particularly in relation to categorization as LBW. This methodology can serve as a tool to monitor various developmental programs aimed at improving birth weights. PMID- 9332115 TI - Antipyretic therapy. PMID- 9332116 TI - NICU environment--a need for change. PMID- 9332117 TI - MCH indicators in south Asia. PMID- 9332118 TI - Cystometric evaluation of voiding dysfunctions. PMID- 9332119 TI - Neonatal jaundice: an analysis of 551 cases. PMID- 9332121 TI - Helium-neon laser as an adjunctive modality for wound healing. PMID- 9332120 TI - Filarial lymphadenitis. PMID- 9332122 TI - Retrobulbar pseudotumor as a manifestation of Staphylococcal pyemia. PMID- 9332123 TI - Multifocal cystic bone tuberculosis with lupus vulgaris and lymphadenitis. PMID- 9332124 TI - Measles vaccination: is the EZ strain safe? PMID- 9332125 TI - Vaccination programme for Japanese encephalitis. PMID- 9332126 TI - HIV and breastfeeding--an alternative. PMID- 9332127 TI - Acute toxicity of vitamin A administered with measles immunization. PMID- 9332128 TI - Intravenous magnesium sulfate: a new weapon against acute asthma? PMID- 9332129 TI - Bone marrow transplantation for sickle cell anemia: is it the right choice? PMID- 9332130 TI - Peptic ulcer in Greenland Inuit: evidence for a low prevalence of duodenal ulcer. AB - In an eight years period new peptic ulcer was diagnosed in 127 Inuit patients at the Central Hospital, Dronning Ingrids Hospital, Nuuk/Godthab. The ratios: duodenal ulcers (DU): prepyloric ulcers (PPU): gastric ulcers (GU) were 17:22:88. The male:female ratio was 2:1. 46 of the patients were living permanently in Nuuk, 81 in The Districts. There were no significant differences in the type of ulcers among the two groups. The incidence of GU among the Nuuk population was comparable to the incidence in the Danish population (0.63/1000 inhabitants per year), whereas the mean age at the time of diagnosis was only 45 years, thus the patients were approximately 15 years younger than the Danish counterparts. The incidence of DU among the Inuits was 0.15/1000 inhabitants per year, significantly less than in the Danish population. The frequency of Helicobacter (H.) pylori infection among 56 Inuits with dyspeptic symptoms was: 0.61. Only 6/12 patients suffering from DU had a positive test for H. pylori infection. CONCLUSIONS: The incidence of duodenal ulcers in the Inuit population was only 10% of the incidence in a Danish population, whereas the incidence of gastric ulcers among the Inuits was comparable to the incidence among Danes. Only 50% of Inuit patients with proven DU had a positive test for H. pylori infection, whereas the frequency of H.pylori infection in a population with dyspeptic symptoms corresponded very well to the frequency reported from other populations. PMID- 9332131 TI - Chlamydia pneumoniae and Helicobacter pylori infections in Sami and Finnish reindeer herders. AB - The Sami people in northernmost Finland have lower mortality from cardiovascular diseases than the main population of Finland. Chronic infections caused by Chlamydia pneumoniae and Helicobacter pylori, both quite recently discovered gram negative bacteria, have been associated with atherosclerosis. We studied the prevalence of these infections in Sami and Finnish men by analysing the C. pneumoniae and H. pylori specific serum IgG and IgA antibodies using microimmune fluorescence and enzyme linked immunosorbent assay methods, respectively. The frequency of C. pneumoniae IgG antibodies and the age adjusted geometric mean titres differed significantly between these groups. The Finns were more often sero-positive than the Sami (76% vs. 67%, respectively), the age adjusted geometric mean titre being 71.6 in the Finns and 38.3 in the Sami; p = 0.001. No significant difference was found in the H. pylori IgG and IgA antibody prevalences, nor in the geometric mean titres between these groups. The difference in cardiovascular mortality between the Sami and Finns may be partly explained by the lower incidence of chlamydial infections in the Sami. PMID- 9332132 TI - The state of young adults with juvenile onset diabetes. AB - Childhood diabetes is most common in Nordic countries and its incidence is rising. In order to evaluate the efficacy of health care follow-up units we investigated physical and psychosocial health status, mode of coping with adult health care and medical treatment in 82 young adults (46 males, 36 females, average age 20.9 yr. and average disease duration 12.7 yr.) who had had diabetes since childhood. All but three of them made regular visits to a health care facility but only 27% monitored blood glucose reasonably well. Only eight percent had a HbA1 concentration within the optimal range, and half had a inappropriate level. Half of the subjects with high HbA1 in adolescence had managed to improve it since leaving the paediatric unit. The most common clinical findings were lipohypertrophy and depressed patellar and achillar reflexes. Up to 70% had background retinopathy and 10% proliferative retinopathy, while two thirds (62%) had depressed conduction velocity of the peroneal nerve. Clinically significant psychiatric problems were found in 17% of the patients, depression being the most prominent feature. Among the social characteristics, delayed social maturation and lack of vocational education were found to be more common than in age-matched controls. One in three exhibited a major overt physical problem and one in five a major psychosocial problem. In conclusion, whatever the health care follow-up unit attended by young adults with diabetes since childhood, the teams face health problems that differ totally from one individual to another. It is important at this transitional age to focus attention in a broad-minded manner on the many factors complicating diabetes or affecting good compliance with treatment. PMID- 9332133 TI - Parental rearing: a comparison between juvenile delinquents and controls in Russia. AB - Since parental maltreatment is commonly reported in studies of juvenile delinquency, our study aimed at a systematic investigation of perceived parental rearing by means of EMBU questionnaire in young male criminals (N = 190) compared to a matched control group (N = 120) in Russia. The results revealed significant differences between delinquents and non-delinquents in terms of the experience of more rejection, abuse, punishment and lack of emotional warmth in the former group. It became obvious that many of the differences were gender-specific, suggesting the crucial role of the fathers in the rearing practices towards their offsprings. The necessity of further study on the interrelationship between family variables and environmental correlates in an interactional model of delinquency is pointed out. PMID- 9332134 TI - Asthma-induced hospitalizations among conscripts in Finland in 1982-1992. AB - The purpose was to examine changes in the numbers of asthma-related hospitalizations among conscripts and possible seasonal fluctuations. Data on treatment periods for asthma among men aged 18-22 years at military hospitals in 1982-1992 were collected from the national hospital discharge register. Monthly numbers of hospitalizations were calculated for each year separately, together with the frequency of such periods per 1000 conscripts. Results. A total of 4894 asthma-related hospitalizations were recorded in 1982-1992, the frequency per 1000 conscripts increasing from 8.5 in 1982 to 27.7 in 1992. Evident seasonal fluctuations were observed in 1982-1989, the peaks being recorded for February (14% above the annual average), July (26%) and November (51%). A change in these seasonal fluctuations was observed in 1990-1992, however. The frequency of asthma induced hospitalizations among conscripts tripled between 1982 and 1992, evidently indicating a real increase in the number of occurrences. The hospitalization peaks are located at the beginning of military service, a point at which factors tending to aggravate asthma exercise a major impact. PMID- 9332135 TI - Much ado about nothing. PMID- 9332136 TI - Treatment of the HIV-infected patient. PMID- 9332137 TI - Nitrous oxide. PMID- 9332138 TI - HIV Postexposure Prophylaxis Registry. PMID- 9332139 TI - Issues related to single-tooth implants. AB - Many practitioners place single-tooth implants when the teeth adjacent to the edentulous space are intact, free of caries and have had no restorations, and the patient is reluctant to wear a removable appliance for physical or psychological reasons, or both. The main exclusionary factors for a single-tooth dental implant are insufficient bone quantity, poor bone quality and insufficient vertical interarch space to accommodate the prosthodontic components of the implant. PMID- 9332140 TI - Clinical features of early-onset periodontitis. AB - The authors estimate the prevalence of early-onset periodontitis, or EOP, in U.S. adolescents and describe the clinical features that occur at an early stage in those who have EOP. In 1986 and 1987, about 10.0 percent of African-American, 5.0 percent of Hispanic and 1.3 percent of white U.S. adolescents had EOP. Clinical features that may be useful in the early detection of EOP include overt gingival inflammation, dental calculus and a high rate of caries, restorations and tooth loss. PMID- 9332141 TI - The efficacy of caries detection using three intraoral films under different processing conditions. AB - This study compares the diagnostic accuracy of caries detection using Ultra-speed (Eastman Kodak), Ektaspeed (Eastman Kodak) and Ektaspeed Plus (Eastman Kodak) films after they were developed in both new and used processing solutions. Ektaspeed Plus film provided significantly better diagnostic accuracy for small proximal-surface caries limited to the outer third of the dentin than did Ektaspeed film. Ektaspeed Plus film did not differ significantly from Ultra-speed film in aiding in the diagnosis of small carious lesions, and it maintained diagnostic accuracy in used processing solutions. Dentists can offer the X-ray dose-reducing technology of Ektaspeed Plus film to their patients and maintain consistently high diagnostic quality. PMID- 9332142 TI - Alcoholic parotid sialadenosis. AB - Alcoholism is a primary cause of sialadenosis, which is an asymptomatic, bilateral enlargement of the parotid glands. The authors outline the pathogenesis, symptoms and testing involved in diagnosing sialadenosis. Recognizing sialadenosis is important because it may point to the unsuspected presence of underlying systemic disease. Therefore, dental practitioners need to be able to differentiate sialadenosis from an inflammatory or neoplastic process to prevent unnecessary treatment. PMID- 9332143 TI - Oral health problems and use of dental services among HIV-infected adults. Supplement to HIV/AIDS Surveillance Project Group. AB - Between January and May 1994, a 14-question survey regarding oral symptoms and use of dental care services was added to a multistate interview project of adults infected with the human immunodeficiency virus. Results indicate that there are disparities and perceived barriers among HIV-infected adults seeking and receiving dental care. Improved dental care services for all HIV-infected people should include better patient and provider awareness of HIV-related oral conditions, more-affordable treatment and expansion of dental insurance coverage. PMID- 9332144 TI - Tinnitus improvement through TMD therapy. AB - Many patients with temporomandibular disorder and coexisting tinnitus find that therapy improves or resolves their tinnitus in conjunction with their TMD symptoms. Ninety-three patients with TMD who reported having coexisting tinnitus were questioned and given clinical tests. These assessment instruments were then evaluated for their ability to suggest tinnitus improvement as a result of TMD therapy. This study suggests that asking targeted questions and performing a clinical test could be of significant value in helping practitioners identify which patients with TMD and coexisting tinnitus will experience improvement in, or resolution of, their tinnitus when TMD symptoms have improved significantly. PMID- 9332145 TI - An alternative for making master impressions for complete dentures. PMID- 9332146 TI - The lingual barbell: a new etiology for the cracked-tooth syndrome. PMID- 9332148 TI - Benefit-to-risk ratio: the challenge of antibiotic drug resistance. PMID- 9332147 TI - Making Medicaid child dental services work: a partnership in Washington state. AB - Eighty-one percent of general dentists and 86 percent of pediatric dentists who are members of the local dental society in Spokane County, Washington, participated in a pilot program to provide dental care in private offices to children up to 5 years of age from low-income families served by the Medicaid program. Outreach staff from the local public health agency recruited and enrolled families in the program. University faculty provided special training in the care of young children to the dentists participating in the program. In the program's first year, 37 percent of the enrolled children had made at least one visit to the dentist, in contrast to 12 percent of children who were not enrolled in the program. PMID- 9332149 TI - Why all-ceramic crowns? PMID- 9332150 TI - The rise of the son (Takao Fusayama). PMID- 9332151 TI - Who's minding the store? PMID- 9332152 TI - Emergency medical services for rape victims: detecting the cracks in service delivery. AB - Rape victims have many emergency medical needs, yet there has been very little research examining whether victims are receiving desired assistance. In this study, 147 rape victim advocates were interviewed about their most recent case in which a victim has sought treatment in an emergency room (ER). The results of this study indicated that there is some inconsistency in which services victims receive and that many women did not obtain the resources they wanted. Some forms of assistance were not provided to victims due to lack of resources in their communities (e.g., follow-up medical care). Other services were not offered due to problems in the implementation of available services (e.g., the morning-after pill [ethinyl estradiol-norgestrel] to prevent pregnancy). This research also examined how characteristics of the hospitals, the assaults themselves, and the victims impacted whether women would receive desired help. These findings indicated that women who were treated in hospitals affiliated with the Catholic church; those who were raped by their friends, dating partners, or husbands; those who experienced multiple forms of forced penetration (vaginal rape and anal rape, oral rape, or rape by an object); women of Color; and victims who did not present a sympathetic demeanor in the ER were less likely to receive several forms of assistance, such as treatment of physical injuries, arranging follow-up medical care, information and treatment for sexually transmitted diseases, information on the risk of pregnancy, the morning-after pill, and information on the physical and psychological health effects of sexual assault. Victims who were taken to hospitals that had coordinated response teams (e.g., Sexual Assault Response Teams) to work with survivors were more likely to receive some forms of treatment (e.g., information on the physical and psychological health effects of sexual assault). Implications for future research and policy initiatives in women's health are discussed. PMID- 9332153 TI - Psychosocial, behavioral, and health factors related to menopause symptomatology. AB - Despite wide variation in the reporting of hot flashes and night sweats among menopausal women, what differentiates symptomatic from asymptomatic women is not well understood. In this article, we use longitudinal data from a large cohort of initially premenopausal women to address premenopausal factors predictive of length of the perimenopause, frequency of hot flash/night sweat (HF/NS) reporting, bothersomeness of HF/NS, and treatment seeking during menopause. The sample for analysis consists of 454 women from the Massachusetts Women's Health Study who were premenopausal at baseline and postmenopausal by the sixth and last study follow-up. Each of the four study outcomes was modeled as a function of premenopausal characteristics using logistic regression. Results confirm a wide range of symptom reporting, with 23% of women not reporting HF/NS at any of the six interviews. Variables related to greater frequency of HF/NS reporting included a longer perimenopause, more psychological and physical symptoms prior to menopause, lower education, and more negative attitudes toward menopause prior to menopause. Symptom bothersomeness was related to greater frequency of HF/NS reporting, smoking, and being divorced. Variables that predicted medical doctor consultation were greater frequency and bothersomeness of symptoms, higher education, and greater health care utilization. We conclude that general symptom reporting, attitudes toward menopause, and lifestyle factors can explain some of the individual variation in symptom reporting. PMID- 9332154 TI - Patterns and correlates of hormone replacement therapy use among middle-aged Australian women. AB - In this study, we examine the patterns of use of hormone replacement therapy (HRT) among women age 51 to 60 years and describe the characteristics of women who currently use HRT, previously used HRT, and have never used HRT. A brief postal survey of 800 women in this age range was used to determine HRT status. Telephone interviews were then conducted with 258 women (111 currently using HRT, 47 who previously used HRT, and 100 who had never used HRT) to determine characteristics of women who currently or previously used HRT or never used HRT, type of HRT used, duration of use, and reasons for use and nonuse. Nearly 40% of women were currently using HRT, 14% had previously used HRT, and 47% had never used HRT. Women currently using HRT were more likely than those not using HRT to have had a hysterectomy, attribute a greater number of symptoms to the climacteric, be in paid employment, and report a greater number of visits to the doctor over the past 12 months. HRT use among Australian women in their 50s is high and rising. Hysterectomy status, the attribution of symptoms to menopause, paid employment, and health care use were the most important correlates of HRT use. Few women specified long-term prevention of osteoporosis or heart disease as a reason for taking HRT. PMID- 9332155 TI - Gender and ethnic differences in readiness to change smoking behavior. AB - The Transtheoretical Model has been used extensively to investigate smoking behavior. However, gender and ethnic differences in key constructs of the Transtheoretical Model have not been fully evaluated. This gap in the literature is addressed in this brief report. We examined gender and ethnic differences in stages of change (readiness to quit smoking), perceived pros (benefits) and cons (costs) of smoking, and self-efficacy (confidence) in ability to quit among smokers seeking cessation treatment. Participants were 330 smokers ages 18 to 75, who responded to advertisements for a free minimal-contact smoking cessation program. Thirty percent of women were confident they could quit smoking compared to 53% of men. Women reported more pros of smoking and more cons of smoking than men. White smokers reported more pros of smoking than African smokers. These findings highlight the need to bolster quitting confidence among women and to identify alternatives to the pros of smoking relevant to women smokers. PMID- 9332156 TI - Cardiovascular reactivity among hostile men and women: the effects of sex and anger suppression. AB - This study examined cardiovascular reactivity differences among hostile men and women. Sixty-four individuals (33 women 31 men; M = 19.9 years of age) were selected from a sample of 105 volunteers based on their Cook-Medley Hostility Scale scores (Cook & Medley, 1954; less than or equal to 24). Analyses revealed no significant sex differences in Cook-Medley Hostility scores. At baseline, men had higher mean systolic blood pressure (SBP) level. However, during the Stroop Color-Word Conflict Task (Stroop, 1935), high-hostile men and women exhibited similar cardiovascular responses. Further analyses revealed that cardiovascular responses to the Stroop task were differentially associated with among men and women as a function of anger suppression. For women, anger suppression was positively associated with diastolic blood pressure (DBP) responses and negatively associated with SBP responses. In contrast, anger suppression was negatively associated with DBP changes and not associated with SBP responses for men. The results suggest that personality factors, such as high hostility and anger suppression, may influence the degree to which men and women differ in their cardiovascular responses to interpersonal stressors. PMID- 9332157 TI - A study on lung toxicity of respirable hard metal dusts in rats. AB - The aetiology of hard metal lung disease has not been clarified so far. The pulmonary toxicity of respirable dusts collected in a hard metal factory was studied in vivo in rats. The effect of the samples was examined 1, 4, 7 and 30 days after single intratracheal injection. Lactate dehydrogenase (LDH), acid phosphatase (AP), protein and phospholipid were determined in cellfree bronchoalveolar lavage (BAL) and lung tissue. The lungs and regional lymph nodes were processed histologically. Lung toxicity of the samples collected during hard metal production varied. Samples containing considerable amount of cobalt dissolved upon acid treatment were found to induce inflammation. It has been established that the biological effect of samples of identical composition is changed by heat treatment and pre-sintering. Our examinations seem to prove that cobalt plays a prominent role in the development of pathological alterations. PMID- 9332158 TI - European sampling intercomparisons for aromatic and chlorinated hydrocarbons in workplace air. AB - Thirty-eight laboratories of the EU Member States, representing government, manufacturers of personal samplers, industrial and university laboratories have participated in a quality assurance scheme which allows to evaluate errors associated with both the sampling and analytical step of personal sampling methods. State-of-the-art bias, within and between laboratory coefficients of variation for pumped and diffusive methods currently applied are discussed. The data enable verification of compliance of the method-laboratory combinations with EN 482 and quantification of errors, in specific related to the sampling step. The merits of the project regarding improved procedures and results are discussed in detail. PMID- 9332159 TI - A comparison of exposures to refractory ceramic fibres over multiple work shifts. AB - As part of an ongoing, industry-wide study in the manufacture of refractory ceramic fibres (RCF), time weighted average (TWA) exposures have been collected at five facilities according to a standardised protocol. Work activities were grouped into dust zones (DZs). Persons to be sampled were randomly selected according to a protocol designed to assure that at least one sample was collected annually from each DZ; each work shift is also sampled at least annually. TWA exposures calculated over a sampling period of at least 360 min were included in the data set. DZs were combined into one of three groups (DZGs): fibre production; vacuum processes; other. The data were analysed to identify any differences by DZG between airborne fibre exposures, by the shift worked at each facility, and across all facilities. There were no statistically significant shift-related differences detected between airborne fibre exposures across the five RCF facilities when analysed as a group. Within four of the facilities, no shift-related differences were detected between airborne fibre exposures; however, at one facility, first and third shift exposures were statistically different. No documentation related to job activities was found to account for the observation. The data generally support the use of a single exposure estimate for each DZG in each of these facilities, regardless of shift worked. Researchers reconstructing exposure and not able to determine the shift worked by study subjects may find these results useful, but are cautioned that substantial differences in exposure across shifts may exist in other types of manufacturing. PMID- 9332160 TI - Quantification of historical dust exposures in the diatomaceous earth industry. AB - Quantitative estimates of dust exposure in a diatomaceous earth (DE) mining and milling operation have been derived based on air sampling records for the period 1948-1988. A total of 6395 records was included in the analysis. Conversion of results obtained by particle counting, expressed as millions of particles per cubic feet (mppcf) of gravimetrically from a filter cassette and expressed as mg m-3 total, were converted to mg m-3 respirable dust using a conversion factor derived from data obtained during the same periods at the plant. Conversion factors were calculated as the average difference of means on the log scale in order to provide stable and consistent conversions and as a ratio of arithmetic means so that the results could be compared with similar studies. After converting the available data to mg m-3 respirable dust, geometric mean (geometric standard deviation) concentrations were 0.37 (2.43) during the 1950s and 0.17 (2.35) during later periods. Exposures were estimated using two linear models, one estimating the changes in concentration over time, and the other providing job-specific mean exposures during the more recent period. Extrapolation of the estimates to periods prior to the availability of any data was done using a subjectively-determined scaling factor. The average estimated respirable dust concentrations for 135 jobs were 3.55 (+/-1.25), 1.37 (+/-0.48), 0.47 (+/-0.16) and 0.29 (+/-0.10) mg m-3 prior to 1949, 1949-1953, 1954-1973 and 1974-1988, respectively. Despite the limitations of the available data, the estimation procedures used are expected to provide reasonable quantitative estimates of silica-containing dust exposure for subsequent exposure-response analyses. PMID- 9332162 TI - Sex determination and dosage compensation in Drosophila: essential components of the hierarchy. PMID- 9332161 TI - Testing fibre release from insulation products: report on a workshop. PMID- 9332163 TI - Lipoproteins: their potential as endogenous target oriented novel drug delivery system. AB - Targeting of drugs to specific cellular sites is a major area of interest now a day. Various carrier systems, viz, liposomes, resealed erythrocytes, niosomes, immunomodulated nanospheres, etc. have been reported. A newer class of carrier system based on endogenous origin has been identified as lipoproteins. The present review discusses the classification, biological fate and targeting potentials of such system when the drug is encapsulated. Furthermore their is a brief discussion about the approaches utilised for their more specific targeting (acyloglycoprotein receptor etc.). Some of the observed drawbacks of such systems could be combated with the use of synthetically prepared lipoproteins named as SMBV (supramolecular biovectors) have also been discussed. Various biomedical potential of these lipoproteins in the delivery of drug to neoplastic cell lines, viral and parasitic infections in liver have also been highlighted. PMID- 9332164 TI - Cytosine methylation inhibits expression of hsp-CAT gene in Drosophila cells. AB - Chloramphenicol acetyl transferase gene under control of hsp promoter (hsp-CAT gene) was introduced and expressed upon heat shock in Drosophila cells at 48 and 72 hr following transfection. Expression of CAT gene was remarkably reduced when DNA methylated at CpG sites was used although presence of methylated plasmid DNA could be demonstrated in cells at 48 and 72 hr. Thus, in Drosophila cells exogenously introduced methylated DNA is expressed differently from unmethylated DNA. PMID- 9332165 TI - Effects of stress on gamma glutamyl transpeptidase (GGT) activity in lymphoid system of rats: modulation by drugs. AB - Effects of stress and its modulation by adaptogens were evaluated on gamma glutamyl transpeptidase (GGT) activity in different tissues of the lymphoid system in rats. Restrain stress (RSx5) suppressed the GGT activity in different tissues of lymphoid system viz. the lymphocyte, the spleen, the thymus and the macrophage, and the maximum effect was seen in the spleen. Chlordiazepoxide, a prototype anti-stress agent, which did not alter GGT activity per se, reversed the effect of RS on this enzyme activity in tissues of lymphoid system studied. Azardirachta indica (Al, Neem), an indigenous adaptogen stimulated the GGT activity per se and nearly normalised RS induced suppression of GGT in lymphoid system. The observed suppression of GGT activity in lymphoid system by stress and its modulation by natural and synthetic adaptogens indicates that GGT could be a consistent cellular/biochemical marker of stress responsiveness and stress induced immunomodulation. PMID- 9332166 TI - ELISA for detection of dengue virus-induced cytokine and its antibody. AB - ELISA technique has been standardized for detection of a dengue type 2 virus (DV) induced cytokine, the cytotoxic factor (CF) and CF-specific antibodies. The performed ELISA was found to be sensitive (90.9%), specific (92.5%), accurate (91%) and reproducible and was able to detect a minimum of 7 ng/ml CF in the test samples. The technique was used to detect CF in DV inoculated mouse sera and DV infected mouse spleen cell culture fluids. Significant utility of the test was detection of a CF-like cytokine in the sera of human cases of dengue haemorrhagic fever. PMID- 9332167 TI - Prevention of radiation induced taste aversion in rats. AB - Diltiazem, a calcium channel blocker, and a cardiovascular therapeutic agent offers significant protection to mice against lethal dose of ionizing radiation. Considering the potential efficacy of diltiazem as a radioprotector for human use, it was deemed necessary to investigate its influence on radiation-induced behavioural changes like nausea, vomiting, learning, memory and performance. In the present studies, conditioned taste aversion (CTA) test based on consumption of saccharin solution, was used as a marker of behavioural changes. Significant CTA (97 +/- 2%) was observed in rats irradiated with Co-60 gamma rays (absorbed dose 1 Gy). Administration of diltiazem at doses greater than 10 mg/kg, body wt, evoked CTA in a dose-dependent manner and that was found to be further aggravated on irradiation. At a lower dose of 5 mg/kg, body wt, diltiazem did not evoke CTA and protected against radiation induced aversion significantly (62 +/- 3%). The results suggest that diltiazem at concentrations lower than 10 mg/kg, body wt, in rats may be useful in preventing radiation induced behavioural changes. This observation could be of particular significance in clinical radiotherapy where radiation induced nausea and vomiting are of great concern. PMID- 9332168 TI - Antioxidant activity of glycowithanolides from Withania somnifera. AB - Antioxidant activity of active principles of Withania somnifera, consisting of equimolar concentrations of sitoindosides VII-X and withaferin A, was investigated for their effects on rat brain frontal cortical and striatal concentrations of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX). Results were compared with effects induced by deprenyl, an agent with well documented antioxidant activity. Active glycowithanolides of W. somnifera (WSG) (10 and 20 mg/kg, i.p.), administered once daily for 21 days, induced a dose-related increase in SOD, CAT and GPX activity in frontal cortex and striatum, which was statistically significant on days 14 and 21, except with the lower dose of WSG on GPX activity, where the effect was evident only on day 21. The data were comparable to those induced by deprenyl (2 mg/kg/day, i.p.) with respect to SOD, CAT and GPX activities, which were evident by day 14. These findings are consistent with the therapeutic use of W. somnifera as an Ayurvedic rasayana and medhyarasayana. Antioxidant effect of active principles of W. somnifera may explain, at least in part, the reported antistress, immunomodulatory, cognition-facilitating, anti-inflammatory and anti-aging effects produced by them in experimental animals, and in clinical situations. PMID- 9332169 TI - Involvement of adrenal hormones in tissue respiration of sub-tropical hibernating and non-hibernating species of frogs. AB - Effects of norepinephrine (NE), epinephrine (EP), corticosterone and cortisol were studied both in vivo and in vitro on the rate of oxygen consumption of tissues (liver, skeletal muscle and kidney) of sub-tropical Indian frogs Rana limnocharis (a hibernating species) and Rana cyanophlyctis (a non-hibernating species) exposed to natural climatic conditions during winter and summer/rainy seasons. Further, the effects of NE and EP were also studied in vitro in the presence of specific beta- and alpha-adrenergic antagonists (propranolol and prazosin). NE, EP and corticosterone, when administered in vivo or in vitro, significantly stimulated the respiratory rate of the tissues of both the species irrespective of the seasons/temperature. Results suggest that NE, EP and corticosterone are directly involved in regulation of the energy metabolism of both hibernating and non-hibernating species of sub-tropical frogs. The calorigenic action of NE and EP seems to be mediated by both beta- and alpha adrenergic receptors. However, the temporal involvement of beta- and alpha adrenergic receptors seems to be tissue-dependent. PMID- 9332170 TI - Effects of silymarin on UDP-glucuronic acid and glucuronidation activity in the rat isolated hepatocytes and liver in relation to D-galactosamine toxicity. AB - Influence of silymarin on UDP-glucuronic acid (UDPGA) and glucuronidation activity of freshly isolated rat hepatocytes in suspension and in rat liver in vivo was examined. Viability of the hepatocytes (> 85%) was not altered in Hank's balanced salt solution at least for 4 hr at 37 degrees C under oxygen. Silymarin at 0.4 mM depleted UDPGA by more than 60% at the end of 4 hr of incubation, the fall in nucleotide pool was rapid and concentration (0.1-0.4 mM)-dependent. The rate of glucuronidation of 3-OH- benzo(a)pyrene (3-OH-BP) determined simultaneously was also reduced significantly; silybin being 3-times more effective than silymarin. Combination of flavonoids with D-galactosamine (GalN) further attenuated the glucuronidation functions of the cells. The flavonoids also offered strong inhibition of UDP-glucose dehydrogenase (UDP-GDH) activity in the liver cytosolic fraction while the activity in hepatocytes was not affected even after 4 hr of incubation. Interestingly, the GalN- induced strong inhibition of UDP-GDH in isolated hepatocytes was completely abolished by flavonoids. Decrease in UDPGA appeared neither due to the activation of UDPGA-pyrophosphatase activity nor to the inhibition of UDP-GDH activity in hepatocytes. Further, the flavonoids also inhibited hepatic UDP-glucuronyltransferase activity towards 3-OH BP (UGT) both in vitro and in intact cells. On the contrary, silymarin administered (70 mg/kg body wt; i.p.) to rats for 3 hr increased the hepatic UDPGA by 2-fold while GalN (400 mg/kg body wt) reduced the nucleotide content to 50% of control. Coadministration of silymarin and GalN restored the UDPGA content significantly while the activities of UDP-GDH and UGT were comparable to the untreated control. The results indicated that silymarin elicits differential effects on the rate of glucuronidation and contents of UDPGA in the isolated rat hepatocytes and in liver. The flavonoid counteracted D-GalN-induced lowering of UDPGA presumably by relieving UDP-GDH of in vivo inhibition affected by GalN metabolite. PMID- 9332171 TI - Effect of vitamin C supplementation on oxidative stress in experimental diabetes. AB - Alloxan diabetic rats supplemented with vitamin C (ascorbic acid) orally in drinking water had increased plasma and liver ascorbic acid as compared to unsupplemented diabetic rats. The levels of liver reduced glutathione also increased in vitamin C supplemented diabetic rats as compared to non-supplemented diabetic rats. Vitamin C supplementation did not have any effect in reducing increased liver lipid peroxidation in diabetic rats. The results of the present study suggest that diabetes results in decreased levels of protective antioxidant species and vitamin C is effective to some extent in maintaining levels of plasma and liver ascorbic acid and liver reduced glutathione. PMID- 9332172 TI - Factors influencing induction and reversal of virus induced diabetes. AB - Injection of encephalomyocarditis virus -D strain in SJL/J mice leads to development of diabetes. In order to ascertain various factors involved in this process, effect of age of the host, dose of virus and glucose pretreatment on incidence of diabetes and its possible reversal were studied. Blood and urine glucose levels of control and experimental mice were followed for 6-8 weeks to reveal diabetic and reversal from diabetic state. It is observed that incidence of diabetes is directly proportional to the age of the host and dose of the virus, leading to maximum destruction of beta cells and minimum chances of recovery from the diabetic state. Glucose injection prior to low dose virus inoculation reduced the incidence of diabetes and enhanced the process of reversal of diabetes. The data reveal the importance of age of host, dose of virus, metabolic state of beta cells and residual beta cell mass in recovery and reversal of virus induced. PMID- 9332174 TI - Responses of aging brain to physical exercise: changes in the substrates of energy metabolism. AB - Responses of aging brain to physical training was evaluated by quantifying the substrates, glucose, lactic acid, and nucleic acids in cerebral cortex (CC) and medulla oblongata (MO) of the brain in rats. Rats of 1 month (young), 6 months (adult), 12 months (middle-aged) and 18 months (old) of age were swim-trained for 30 days. Glucose content of CC and MO increased with training whereas blood glucose decreased in trained young and adult animals with middle-aged and old animals maintaining constant blood glucose. Brain lactate in these two regions decreased with training in all age groups. However, the old animals showed an elevation in blood lactic acid in trained state, while the other age groups showed a decrease. Nucleic acid content, decreased with age, especially the RNA content in MO showing a larger depletion. However, there was no discernible influence of physical exercise on these parameters. Physical training has influenced the aging brain's adaptability, as seen by increase in its glucose content in young animals and also possible utilization of lactate as an additional substrate in old animals as evidenced by an increase in blood lactic acid. PMID- 9332173 TI - Plasma protein binding of theophylline in late pregnancy in rat. AB - A single bolus dose of theophylline (Th; 5 mg/kg) was administered through the abdominal catheter in nonpregnant and pregnant rats to measure unbound fraction (fu) of Th in pregnancy. After 90 min blood samples were collected. Plasma was extracted with a solvent system comprising chloroform and isopropyl alcohol (IPA) followed by iso-cratic HPLC analysis. Results showed an increase in unbound fraction of Th in late pregnancy. PMID- 9332175 TI - Location and mapping of spontaneous mutations in Drosophila bipectinata. PMID- 9332176 TI - Hypoglycemic effect of Prunus amygdalus seeds in albino rabbits. AB - On administration of 2.5 g almond seed and its proportionate fractions, viz. 1.22 g defatted seed and 1.28 g oil to three groups of albino rabbits, they showed a definite hypoglycemic action during a two months study. The active factor seems to be a non oil fraction which is only partly soluble in ethyl ether. PMID- 9332177 TI - Effect of Trasina, an Ayurvedic herbal formulation, on pancreatic islet superoxide dismutase activity in hyperglycaemic rats. AB - Diabetes mellitus was induced in male CF strain rats by streptozotocin (STZ) and hyperglycaemia and superoxide dismutase (SOD) activity of pancreatic islet cells was assessed on days 7, 14, 21 and 28. STZ induced significant hyperglycaemia and a concomitant decrease in islet cell SOD activity. Transina (TR), an Ayurvedic herbal formulation comprising of Withania somnifera, Tinospora cordifolia, Eclipta alba, Ocimum sanctum, Picrorrhiza kurroa and shilajit, had little per se effect on blood sugar concentrations and islet SOD activity in euglycaemic rats, in the doses of 100 and 200 mg/kg, p.o. administered once daily for 28 days. However, these doses of TR induced a dose- related decrease in STZ hyperglycaemia and attenuation of STZ induced decrease in islet SOD activity. The results indicate that the earlier reported anti-hyperglycaemic effect of TR may be due to pancreatic islet free radical scavenging activity, the hyperglycaemic activity of STZ being the consequence of decrease in islet SOD activity leading to the accumulation of degenerative oxidative free radicals in islet beta-cells. PMID- 9332178 TI - Antimicrobial potentiality of a new beta-lactum antibiotic fosfomycin. AB - Antimicrobial action of penicillin and some of its derivatives including fosfomycin was studied with respect to 225 strains of Gram-positive and Gram negative bacteria. Fosfomycin was found to possess somewhat less activity against Staphylococcus aureus compared with other penicillins; however, it showed powerful activity towards Escherichia coli, Klebsiella spp. and Proteus mirabilis. PMID- 9332179 TI - Interaction of muscle relaxant effect of gallamine with ethanol: in vivo mammalian model. AB - Ethanol produces alterations in muscular contraction and neuromuscular function both in vitro and in vivo in mammalian and amphibian tissues in dose dependent manner. It is not very well known whether gallamine, a commonly used muscle relaxant produces any interaction with ethanol. Hence in the present study, interactions of ethanol with gallamine are undertaken using rabbit head drop method. The latency time to produce head drop by gallamine in the absence and presence of ethanol (dose which produce ataxia) was insignificantly different (P > 0.05). It may be due to fact that ataxic dose may not be affecting neuromuscular transmission a significant manner. It, therefore, does not warrant to adjust the dose of gallamine in alcoholic persons. PMID- 9332180 TI - Lipid production by Candida Y-1. AB - Ten oleaginous yeasts were isolated from sources like fruits, flowers, curd whey and soil samples. Lipid production varied from 24 to 71.2% of dry weight of cell biomass. Strain Y-1, which was isolated from curd whey, produced the highest amount of lipid. It was identified belonging to the genus Candida. The optimal cultural conditions for lipid production by Candida Y-1 were found to be glucose (5%) as carbon source, ammonium sulfate (1%) as nitrogen source, pH 4.5, and temperature 28 degrees C under shake-flask conditions (125 rpm). PMID- 9332232 TI - Cerebral CT angiography in the diagnosis of acute subarachnoid hemorrhage. AB - PURPOSE: To evaluate the usefulness of CT angiography (CTA) in the detection of intracranial aneurysms in patients with acute subarachnoid hemorrhage (SAH). MATERIAL AND METHODS: In 53 patients with nontraumatic SAH a helical contrast enhanced CTA was performed. CTA data were reconstructed with maximum intensity projection (MIP). Each patient underwent selective arteriography of the cerebral vessels (as the gold standard). CTA (axial images and MIP reconstructions) and arteriography were evaluated separately and their diagnostic information was compared. RESULTS: In 14 of the 53 patients neither CTA nor angiography showed a vascular malformation. In the remaining 39 patients, angiography demonstrated a total of 51 aneurysms ranging in size from 3 mm to 16 mm. CTA missed one of these aneurysms, which was located at the internal carotid artery. 3-D CT reconstruction was slightly superior to arteriography in the demonstration of the neck, shape and direction of the aneurysms. Partial thrombosis of 3 aneurysms was demonstrated only by CTA. CONCLUSION: Although CTA cannot replace cerebral arteriography in the diagnostic work-up of acute SAH, it proved to be helpful in demonstrating the topographic anatomy of cerebral aneurysms and surrounding structures. PMID- 9332233 TI - Angiography in non-traumatic brain haematoma. An analysis of 100 cases. AB - PURPOSE: The primary purpose of this project was to study the anatomical characteristics of intracerebral haematoma (ICH) in order to determine features that may negate the need for angiography in some patients. MATERIAL AND METHODS: The study was prospective and designed to investigate the underlying cause of non traumatic ICH in 100 cases assessed by conventional angiography. Patients were excluded if there was a history of trauma or known pre-existing brain abnormality. All patients were examined with CT and angiography within 4 days of the ictus. RESULTS: Ruptured aneurysms or arteriovenous malformations (AVMs) were diagnosed on the initial angiogram in 49% of cases: 27 AVMs and 22 aneurysms. One case of superior sagittal sinus thrombosis was also detected. Vascular abnormalities were found most frequently in the under-40 age group and in cases in which subarachnoid haemorrhage, intraventricular haemorrhage or extracerebral haematoma accompanied the ICH. The temporal lobe was the most frequent anatomical location (37%). When a temporal lobe haematoma extended into the Sylvian fissure from the inferior pole of the temporal lobe or when it was associated with subarachnoid haemorrhage, structural abnormalities were found in over 90% of cases. CONCLUSIONS: There are groups of patients with ICH in whom the CT features are highly suggestive of AVM or aneurysm rupture. If the initial angiography is negative in these cases, careful follow up by repeat angiography and/or MR imaging is essential. However, potentially treatable abnormalities cannot be excluded with certainty by the distribution of the haematoma on CT alone, even if there is a history of pre-existing hypertension. PMID- 9332234 TI - Craniocaudal motion velocity in the cervical spinal cord in degenerative disease as shown by MR imaging. AB - PURPOSE: To investigate, by means of MR phase imaging, the effects of compression on the velocity of craniocaudal motion in the spinal cord. MATERIAL AND METHODS: Spin-echo pulse sequences with velocity encoding gradients were used to examine 12 patients with cervical spondylosis and 6 normal volunteers. Oblique-axial phase images at 3 levels (cranial, middle and caudal), were obtained with prospective electrocardiogram gating. The middle level was set at the site where the spinal cord was most severely compressed, and the cranial and caudal sections were set where it was not compressed. Time-velocity curves were generated at these 3 levels and focal velocity change was correlated with motor function in the lower extremities. RESULTS AND CONCLUSION: The cord showed a higher motion velocity at the compression level than at noncompression levels. This paradoxical increase in velocity was observed in 7 out of 8 patients whose lower extremity motor function was impaired. Four patients with normal lower extremity motor function did not demonstrate this increase in velocity. An increase in motion velocity was therefore found to correlate with impaired lower extremity motor function. PMID- 9332235 TI - MR imaging in squamous cell carcinoma of the head and neck with no palpable lymph nodes. AB - PURPOSE: To assess the efficacy of MR imaging in the detection of lymph node metastasis in patients with no palpable lymph nodes ("N0 neck") who have squamous cell carcinoma of the head and neck region. MATERIAL AND METHODS: MR neck imagings in 18 patients who underwent neck dissection (bilaterally in 2) for squamous cell carcinoma of the head and neck region were examined preoperatively for the purpose of detecting lymph node metastases. The imaging features taken into consideration were: size (cutoff point 10 mm), grouping, presence of central necrosis, and appearance of extracapsular spread. The MR examinations comprised spin-echo T1- and T2-weighted sequences. The MR findings were compared with those of surgery and histopathological examination. RESULTS: MR suggested metastatic lymph node involvement in 5 necks. In 2 of these, central necrosis was seen in the enlarged lymph nodes. In a third, a grouping of the lymph nodes was noted. Extracapsular spread was not present. Histopathological examination revealed metastatic lymph nodes in 7 of the 20 necks, the rate of clinically occult disease being 35%, and 4 of them had been accurately graded by MR. There was one false-positive MR examination. The MR sensitivity was 57.1% and specificity 92.3%. CONCLUSION: MR may reveal metastatic lymph nodes in patients with no clinical evidence of metastasis. However, conventional MR techniques are not always sufficient for decision-making on surgery in cases of "N0 neck". PMID- 9332237 TI - Transcutaneous needle biopsy of the lung. AB - PURPOSE: To evaluate the usefulness of transcutaneous needle biopsy (TCNB). MATERIAL AND METHODS: From May 1988 to December 1994, we performed TCNB under fluoroscopic control in 408 patients with mass lesions of the peripheral lung. The Surecut needle (1.5 mm) was selected mainly because of its ability to obtain specimens large enough for histological examination. Of the 408 patients, 286 had had previous bronchofiberscopic examinations but no definite diagnosis had been reached. RESULTS: A definite diagnosis was obtained by TCNB in 305 (74.7%) of 408 cases (251 malignant neoplasms, 54 benign lesions). In malignant neoplasms, the pathological diagnosis based on cytology and histology together was more reliable than that based on cytology alone. Although the complications of this procedure (such as pneumothorax) were within the range of acceptability, care should be taken to avoid air embolism and the seeding of cancer cells along the needle tract. CONCLUSION: TCNB with the Surecut needle is a useful procedure with relatively low risk. PMID- 9332236 TI - Solid nonpalpable breast lesions. Success and failure of guided fine-needle aspiration cytology in a consecutive series of 2444 cases. AB - PURPOSE: To evaluate the contribution of guided fine-needle aspiration cytology in reducing unnecessary biopsies of benign solid nonpalpable breast lesions with low suspicion of malignancy at mammography. MATERIAL AND METHODS: An evaluation was made of a consecutive series of 2444 solid nonpalpable breast lesions detected by mammography and undergoing guided (sonography or stereotaxy) fine needle aspiration cytology. Surgical biopsy was made in the presence of strong suspicion of malignancy at mammography and/or of abnormal cytology. RESULTS: The sensitivity was 96.7% and the specificity 77.7% (average follow-up 2.77 years). False-negative/inadequate cytology associated with low suspicion of malignancy at mammography resulted in a diagnostic delay in 27 cancer cases (invasive 20, intraductal 7). On the other hand, cytology led to surgical biopsy in 53 cancer cases which might not otherwise have been biopsied because of low radiological suspicion of cancer. Surgical biopsy of all cases, to avoid diagnostic delays, would have increased the benign biopsy rate by a factor of 4.5, with a rise in the benign: malignant biopsy ratio from 0.44:1 to 1.93:1. CONCLUSION: Stereotaxy- or ultrasound-guided fine-needle aspiration cytology of nonpalpable mammographic abnormalities can achieve a sharp reduction in unnecessary benign biopsies in cases of low suspicion of malignancy at mammography. PMID- 9332238 TI - Progress of emphysema in severe alpha 1-antitrypsin deficiency as assessed by annual CT. AB - PURPOSE: To assess serial CT as a measure of the progress of emphysema in patients with severe alpha 1-antitrypsin deficiency (phenotype PiZ). MATERIAL AND METHODS: In a randomized placebo-controlled study of alpha 1-antitrypsin augmentation therapy, 22 patients with moderate emphysema were followed for 2-4 years with an annual lung CT. The images were analysed by means of semiautomatic lung detection, and the degree of emphysema was quantitated by the density-mask and the percentile methods. The influence of lung volume was standardised by a regression model. RESULTS: A highly significant decline in Hounsfield units (HU) was found in low-density areas, corresponding to a mean (SE) annual loss of lung tissue of 2.1 (0.4) g/l lung volume. Analysis of a single slice at 5 cm below the level of the carina gave comparable results: 2.4 (0.4) g/l. CONCLUSION: Serial CT is a sensitive measure of the progress of emphysema in patients with severe alpha 1-antitrypsin deficiency. PMID- 9332239 TI - Malignant lymphoma arising from chronic tuberculous empyema. A case report. AB - We report on a case of malignant lymphoma in the chest wall, associated with chronic tuberculous empyema. CT and MR imaging showed a soft-tissue mass contiguous with the empyema and invading the chest wall. MR imaging demonstrated a difference in signal intensity between the mass and the empyema. The extent of the chest-wall lymphoma was optimally delineated on fat-suppressed contrast enhanced MR images. PMID- 9332240 TI - Compulsory superselective arterial embolization in hypovascular local hepatic tumor ablation. Microballoon coaxial catheterization. AB - We performed compulsory superselective transcatheter arterial embolization on local hypovascular liver metastases under balloon occlusion using a 1-mm (3 F) coaxial microballoon catheter in 2 cases. One case was a metastasis from breast cancer (maximum diameter 5.5 cm) at segment 7. The other case comprised metastases from rectal cancer (maximum diameter 8 cm) at segments 7 and 8. Absolute ethanol (50%) mixed with Lipiodol (50%) was used for embolization. No major treatment-related complications occurred. No local recurrence was observed in either case in follow-up CT and MR studies of up to 16 and 9 months respectively. This technique may thus be applied as an alternative to surgical resection in the treatment of local hypovascular liver tumors. PMID- 9332242 TI - Radiological findings in serous surface papillary carcinoma of the ovary. Case reports. AB - Serous surface papillary carcinoma (SSPC) is a malignant tumor originating from the surface epithelium of the ovary, and is characterized by macroscopic normal ovaries accompanied by diffuse peritoneal dissemination. Therefore diagnosis of SSPC cannot usually be made preoperatively. We report on two cases of SSPC in which US, CT, and MR demonstrated nodularities along the surface of the normal sized ovaries, adjacent organs and pelvic peritoneum in addition to findings of peritoneal seeding. These imaging findings may suggest the correct preoperative diagnosis of SSPC. PMID- 9332241 TI - Renal vascular resistance in autosomal dominant polycystic kidney disease. Evaluation with color Doppler ultrasound. AB - PURPOSE: The purpose of this study was to evaluate the performance of color duplex Doppler ultrasonography in the assessment of renal vascular resistance (RVR) by measuring resistive indices (RIs) and pulsatility indices (PIs) in patients with autosomal dominant polycystic kidney disease (ADPKD), and to correlate the measured values with renal function and the presence of arterial hypertension. MATERIAL AND METHODS: In 42 patients with ADPKD and 65 control subjects, RIs and PIs were measured by means of color duplex Doppler sonography and correlated with clinical and laboratory findings and with morphological abnormalities at B-mode ultrasonography. RESULTS: Mean RI in the control subjects was 0.59 +/- 0.03 (+/-SD) and in the patients 0.71 +/- 0.11, (p < 0.01). Mean PI in the controls was 1.00 +/- 0.11 and in the patients 1.69 +/- 0.21, (p < 0.01). Elevated RIs and PIs heralded a progression of ADPKD. Doppler indices correlated significantly with renal function tests and morphological changes in the affected kidneys at ultrasound. Significantly higher RIs (p < 0.01) and PIs (p < 0.04) were measured in hypertensive ADPKD patients as compared to normotensive patients. Correlation of patient age and Doppler indices did not reach statistical significance. CONCLUSION: Doppler indices do reflect the increased RVR in patients with ADPKD and they correlate with renal function disturbance, with the development of systemic arterial hypertension, and with ultrasonographic abnormality of the kidney in these subjects. PMID- 9332243 TI - Air enema used in the evaluation of acute colitis. A comparison between instant radiography and endoscopy. AB - PURPOSE: The purpose of this study was to evaluate air enema as a method of assessing acute colitis. MATERIAL AND METHODS: Twenty-seven patients with symptoms of acute colitis underwent plain abdominal radiography, air enema, and colonoscopy within 48 h. The films were evaluated by 3 observers with different levels of experience, both independently and together, and the results were then compared to the findings at endoscopy. RESULTS: Air enema visualized a greater part of the colon than plain abdominal radiography. When air enema was compared to endoscopy as the reference, it showed good correlation, with a positive predictive value of 92% (sensitivity 62%, specificity 85%). Evaluation of the rectum was less accurate, a finding that emphasized the importance of rigid sigmoidoscopy. CONCLUSION: Air enema is a useful diagnostic method in acute colitis, it is easily performed and tolerated well with no observed complications. It is also easy to interpret, as shown by a high level of agreement (kappa = 0.67) among the 3 independent observers with very different levels of experience. PMID- 9332244 TI - Epiphyseal bone-marrow abnormalities and restitution in Legg-Calve-Perthes disease. Evaluation by MR imaging in 86 cases. AB - PURPOSE: By means of MR imaging, to determine signal abnormalities in the femoral epiphysis; to determine their location, extent and restitution over time; and to correlate these findings to the Catterall radiological classification. MATERIAL AND METHODS: A total of 247 MR images in 86 patients (101 hips) with Legg-Calve Perthes disease were examined. The MR images were taken in the coronal plane, and the images through the center of the femoral head were used for this study. RESULTS: T1-weighted images proved as good as T2-weighted images for the MR evaluation of the extent of the necrosis. In almost every case, the central cranial part of the epiphysis showed a low initial signal. In Catterall group I, the medial part was never involved. In Catterall III and IV, almost the entire epiphysis showed signal changes. In the period 3-6 years after diagnosis, we still found signal changes in the epiphysis in some hips but there was no correlation with the Catterall classification. After 6 years, the epiphysis showed normal signal intensity in MR imaging. In T1-weighted images, Gd enhancement occurred in the peripheral regions in the early stages of the disease. The central part of the epiphysis became more enhanced over time and peaked in the period 1-3 years after diagnosis. CONCLUSION: MR is a valuable modality for monitoring changes in the femoral epiphysis. We propose a new classification of the extent and pattern of epiphyseal bone-marrow abnormalities based on the 4 zones most commonly observed in MR imaging. PMID- 9332245 TI - MR findings in cases of suspected impacted fracture of the femoral neck. AB - PURPOSE: To evaluate MR imaging of the hip in patients with a clinically suspected impacted fracture of the femoral neck in cases where conventional plain films show negative or equivocal findings. MATERIAL AND METHODS: Twenty-seven such patients were prospectively examined by MR imaging with a 1.0 T unit, within 24 hours of admittance to hospital. A coronal T1-weighted turbo spin-echo sequence (n = 27), and a coronal STIR sequence (n = 25) or a coronal T2-weighted turbo spin-echo fat saturation sequence (n = 2) were used. The evaluations were made by 2 radiologists with experience in musculoskeletal radiology. RESULTS: There were 6 patients with a pertrochanteric fracture, 2 without and 4 with slight displacement. Five patients had an impacted fracture of the femoral neck, and 3 had a fracture of the superior pubic bone. Of 2 patients with advanced arthrosis, 1 had an impacted femoral neck fracture and the other a nondisplaced intertrochanteric fracture. There was 1 patient who had sustained a nondisplaced acetabular fracture with increased joint fluid and muscle contusions. Three patients had muscle contusions only. Two patients had bone marrow contusions only, while 2 others with advanced coxarthrosis had increased joint fluid only. Three patients showed normal findings. Our findings led to emergency surgery in 13 cases, and conservative measures directed to the specific MR findings in 14 patients. CONCLUSION: MR imaging should be the first modality of choice in examining patients with a clinically suspected impacted fracture of the femoral neck where conventional films show negative or equivocal findings. PMID- 9332246 TI - Sonographic evaluation of hip effusion in children. Improved visualization with the hip in extension and abduction. AB - PURPOSE: To assess the difference in the sonographic appearance of hip effusions when the hip was placed in the extended and abducted position as compared to a neutral position. MATERIAL AND METHODS: Twenty-one consecutive children (aged 2 14 years) with hip pain or limping were examined for hip effusions by means of ultrasound. The capsule-femoral-neck distance, the presence of joint fluid, and the shape of the anterior capsule were compared in hips in slight extension and abduction with those in hips in a neutral position. RESULTS: Of the 11 positive hip effusions, the maximal distance between the capsule and femoral neck was significantly larger (p < 0.005) in the extended and abducted hip position than in the neutral hip position, with a mean difference of 1.6 mm. Convex bulging of the anterior capsule was more confidently diagnosed in 3 hip effusions in the extended and abducted hip, and better visualization of fluid between the capsule and femoral neck was achieved in 4 joint in this posture. CONCLUSION: The study demonstrated an improvement in the sonographic detection of hip effusion in the extended and abducted position compared to the neutral position. PMID- 9332247 TI - CT of the sacroiliac joints. Dosimetry and optimal settings for a high-resolution technique. AB - PURPOSE: The aim was to select an optimal technique for low-dose high-resolution CT of the sacroiliac joints (SJ). MATERIAL AND METHODS: Dose measurements were performed on a Rando anthropomorphic phantom using thermoluminescence dosimeters for 4 CT protocols and 2 conventional radiography protocols used for SJ evaluation. Six available reconstruction algorithms were tested on CT protocols using 285-665 mAs and 120 or 130 kVp settings and noncontiguous 1.5-mm-thin sections with 3.5-mm intervals. Settings with optimum performance on phantom tests were also applied in a series of 10 patients with SJ arthropathies. RESULTS: A CT protocol using 120 kVp/175 mA/2.9 s/1.5-mm slice thickness/5-mm table increment implied the lower radiation dose among all examination protocols tested and provided high image quality of the SJ. A reconstruction algorithm yielding images of improved spatial resolution with acceptable noise was selected. CONCLUSION: A high spatial frequency reconstruction algorithm, and 120 kVp and 508 mAs were considered optimal for a low-dose CT examination of the SJ that employed narrow (1.5 mm) slice images with interspacing. PMID- 9332248 TI - MR imaging of meniscal subluxation in the knee. AB - PURPOSE: The aim of this study was to establish diagnostic criteria for meniscal subluxation, and to determine whether there was any connection between meniscal subluxation and other common meniscal and knee-joint abnormalities. MATERIAL AND METHODS: The normal position of the meniscal body was assessed in 10 asymptomatic volunteers. MR signs of meniscal subluxation were evaluated retrospectively in 60 symptomatic patients with pain in the knee, impaired mobility, and/or joint swelling who had no clear diagnosis after the evaluation of case history, clinical examination, and radiography. The criterion for subluxation of the meniscus was defined as a distance of > or = 3 mm between the peripheral border of the meniscus and the edge of the tibial plateau. RESULTS: In the volunteers, the mean distance from the medial meniscus to the edge of the tibial plateau was 0.07 mm, and that from the lateral meniscus was 0 mm. In 55 symptomatic patients without meniscal subluxation, the mean distance from the meniscus to the edge of the tibial plateau was 0.27 mm. Five patients (8%) had evidence of meniscal subluxation, 4 in the medial meniscus and one in the lateral meniscus. The most commonly associated knee abnormality was joint effusion in 5 knees and osteoarthritis in 2 knees. CONCLUSION: Meniscal subluxation was not a rare finding with MR imaging in patients with painful knees. Meniscal subluxation was associated with other knee abnormalities such as joint effusion or osteoarthritis. PMID- 9332249 TI - GRASE: ultra-fast turbo gradient spin-echo sequence. A new approach to fast MR imaging of the musculoskeletal system. AB - PURPOSE: Ultra-fast gradient and spin-echo (GRASE) imaging is a hybrid of turbo spin-echo (TSE) and echo-planar imaging (EPI). One scan consists of several spin echoes (SEs) (turbo factor, TF), each of which consists of a number of gradient echoes (EPI factor, EF). The aim of our study was to evaluate different combinations of TF and EF in GRASE imaging and to test its usefulness in musculoskeletal imaging. MATERIAL AND METHODS: On a 1.0 T MR unit, 11 GRASE sequences with different combinations of TF and EF (TR/TE 2150/120 ms) were evaluated in phantom studies with respect to signal-to-noise (S/N) ratio, nonuniformity of images, and geometrical distortion. From this study, the optimal GRASE-sequence was applied to 25 patients with different joint pathologies and compared to a T2-weighted TSE sequence (TR/TE 2855/130 ms). Lesion visualization, conspicuity, overall image quality, and artifacts were qualitatively analyzed by two observers independently of each other. RESULTS: With respect to S/N ratio, signal nonuniformity, and geometrical distortion, the GRASE sequence with TF/EF 7/3 (S/N 47; signal nonuniformity 11.7%; distortion 1 pixel) proved to be superior to the other GRASE sequences within a scanning time of less than 120 s. In a clinical study, the GRASE sequence proved superior to T2-weighted TSE (without fat suppression) in the visualization of bone-marrow and soft-tissue lesions (p < 0.001) and ligamentous injuries, although the image quality was inferior. PMID- 9332250 TI - Kinematic MR imaging of the ankle--initial results with ultra-fast sequence imaging. AB - PURPOSE: In order to evaluate the advantages of ultra-fast MR sequences, kinematic MR imaging studies were performed in 4 patients with osteochondritis dissecans of the talus and in 12 healthy volunteers. MATERIAL AND METHODS: The patients and volunteers were placed inside a custom-made positioning device. Sagittal ultra-fast T2-weighted turbo gradient-echo sequences and HASTE sequences were obtained during active joint motion from dorsiflexion to plantar flexion. Eight sagittal slices were scanned separately to cover this ankle motion. In each slice, 8 to 10 images were obtained in 12-s or 18-s periods. RESULTS: Adequate image quality for analyzing the normal kinematics of the ankle was obtained in all subjects. At surgery, the osteochondral fragment was found to be mobile in 3 of the 4 of the patients. In none of these cases was fragment mobility observed on kinematic MR imaging. No motion of the fragments was observed in the fourth patient, neither at surgery nor on kinematic MR imaging. CONCLUSION: Ultra-fast MR imaging sequences made it possible to produce kinematic MR imaging studies of active joint motion. The positioning device was useful for guiding joint motion in patients and for obtaining adequate image quality. PMID- 9332251 TI - Assessment of suspected bone metastases. CT with and without clinical information compared to CT-guided bone biopsy. AB - PURPOSE: To evaluate the role of CT with and without clinical information as compared to CT-guided bone biopsy in the assessment of suspected bone metastases. MATERIAL AND METHODS: The study comprised 51 consecutive patients with suspected bone metastases who had undergone CT-guided bone biopsies with an eccentric drill system. CT of the targets, clinical information, and histopathology were scored separately as malignant, uncertain or benign. The results of CT alone and CT in combination with clinical information were compared to the results of histopathology. RESULTS: Histopathology diagnosed 45/51 lesions (88%), 23 as malignant and 22 as benign. CT correctly depicted 17 of these 23 malignant lesions. The remaining 6 malignant lesions were CT-scored as uncertain (n = 5) or benign (n = 1). CT correctly depicted only 3 of the 22 benign lesions. The remaining 19 benign lesions were CT-scored as malignant (n = 2) or uncertain (n = 17). When uncertain CT scores were combined with clinical scores, the true positive and true-negative results for malignancy increased from 44% to 82%. CONCLUSION: In most cases, CT in combination with clinical information gives enough information about the nature-malignant or benign-of a skeletal lesion. In uncertain cases, diagnostic accuracy can be improved by means of CT-guided bone biopsy. PMID- 9332252 TI - Bone-marrow MR imaging before and after autologous marrow transplantation in lymphoma patients without known bone-marrow involvement. AB - PURPOSE: To study lumbar bone marrow by means of MR imaging before and after bone marrow transplantation in lymphoma patients. Particular emphasis was paid to heterogeneity and to focal manifestations, i.e. appearances that could simulate tumor. MATERIAL AND METHODS: Twenty-two patients who were disease-free for a minimum of 30 months after transplantation were studied in 107 MR examinations. Two radiologists visually evaluated coronal T1-weighted and short inversion time inversion-recovery (STIR) images. RESULTS: T1-weighted images demonstrated a more heterogeneous marrow after transplantation than before it. Sharply defined focal low signal intensity areas appeared on this sequence in 5 (23%) of the 22 patients at between 21 and 60 weeks after transplantation. The mean age of these 5 patients was 48.4 years (range 42-54 years). The difference in age between these 5 patients and the remaining 17 patients, who had a mean age of 33.4 years (range 14-51 years), was statistically significant (p < 0.01, Student's t-test, 2 sided test). CONCLUSION: Sharply defined focal low signal intensity areas may be seen on T1-weighted images of bone marrow in patients who are in complete remission after transplantation, particularly in those aged over 40-45 years. PMID- 9332253 TI - CT, MR imaging and muscle biopsy in severe crush injury. AB - PURPOSE: We reviewed CT and MR findings in crush injuries in muscles and compared them with findings at muscle biopsy. MATERIAL AND METHODS: CT and MR examinations were made of the lower extremities in 6 adult patients whose muscles were crushed in the big earthquake in Kobe, Japan, on 17 January 1995. All CT examinations were performed twice and within 60 days of the earthquake. In 5 of the 6 patients, biopsies were taken of the injured muscle after the CT and MR examinations. RESULTS: At CT investigation, the images in 5 patients showed calcification in the muscles, and the crushed muscle appeared atrophic. In 3 of these 5, the degree of calcification decreased with time. At MR investigation, T1 weighted images of the crushed muscle showed isointensity in 4 of the 6 patients and high intensity in the remaining 2. T2-weighted images of the crushed muscle showed inhomogeneous high intensity in all patients. After Gd injection, all the crushed muscles were enhanced, either slightly or markedly. At muscle biopsy, in 5 of the 6 cases, the findings showed calcification in the muscle cells and the total destruction of the normal muscle structure. CONCLUSION: CT and MR findings in crush injury correlated well with the findings at histopathology. PMID- 9332255 TI - Optimising imaging parameters in experimental spiral CT. AB - PURPOSE: This in vitro study was conducted to analyse lesion detection and relative radiation exposure in different CT techniques. MATERIAL AND METHODS: We used a plastic phantom (12 x 8 x 2 cm) containing holes filled with air or fluid of varying densities to simulate lesions. This was imaged with Siemens Somatom Plus S and GE High Speed Advantage units. We varied table feeds (3 and 6 mm/s in Siemens and 3 and 4.5 mm/s in GE) and increments (2 mm and 4 mm) while keeping collimation at 3 mm. The SmartScan program of GE and the reformatting algorithm of Siemens were also analysed. To evaluate the different methods, the phantom lesions were counted by 3 observers. Radiation exposures associated with each technique were also measured. RESULTS: The images reformatted to a coronal direction were significantly inferior (p < 0.01) to those in other techniques. The use of SmartScan did not influence lesion detection, nor did changes in pitch or increment. Spiral and non-spiral techniques proved to be equal. Radiation exposure was lowest when a greater pitch or the SmartScan program was used. CONCLUSION: Radiation exposure in CT can be limited without significantly impairing the image quality by using low-dose techniques. Reformatting to a coronal direction should be used with care as it debases the image quality. PMID- 9332254 TI - Role of MR venography in the evaluation of deep venous thrombosis. AB - PURPOSE: MR venography has been recommended for the evaluation of deep venous thrombosis. The purpose of our study was to determine the role of MR venography, in particular at the level of the pelvis where other diagnostic modalities show major limitations. MATERIALS AND METHODS: Forty-three patients with clinical suspicion of deep venous thrombosis were examined by means of pelvic MR venography. In all cases, a 2D-TOF sequence was used with cranial arterial presaturation. In selected cases, i.e. when a small intraluminal filling defect was present, a cine-PC sequence was used in addition in order to exclude the presence of a pulsatility artifact as causing the filling defect. In all cases, contrast venography was also performed and considered to be the standard of reference. RESULTS: MR venography showed 26 patients to be positive for deep venous thrombosis at the pelvic level. These positive results were correct in 25 cases. The analysis of the results provided values of sensitivity and specificity of respectively 100% and 94%, with an overall accuracy of 97.6%. CONCLUSIONS: Our results indicate that MR can provide highly accurate images, similar to those of contrast venography, in a noninvasive fashion. It is particularly useful in the pelvic region where the limitations of other imaging modalities are more evident. PMID- 9332256 TI - Permeability of contrast media through hemodialysis membrane. AB - PURPOSE: To compare the permeability of 10 contrast media (CM) through the hemodialysis membrane. MATERIAL AND METHODS: An experimental model was constructed from clinical hemodialysis equipment. The 10 CM that were used were: diatrizoate, iothalamate, iopamidol, iohexol, iopromide, ioversol, iomeprol, ioxaglate, iodixanol and iotrolan. They were dissolved into the dialysate. Each solution ran into the blood circuit of the dialyzer at a rate of 200 ml/min and into the dialysate circuit of the dialysis machine at a rate of 500 ml/min. Permeability was expressed as clearance, calculated from the iodine concentration at the afferent and efferent lines of the dialyzer, and from the blood-flow rate. RESULTS: The clearance of all the monomeric CM was superior to that of all the dimeric CM. Values of clearance and molecular weight correlated well. CONCLUSION: The molecular weight of a contrast medium is the main factor that influences the efficiency of CM removal by hemodialysis. PMID- 9332257 TI - Hidden compartments in intracranial arteriovenous malformation. PMID- 9332258 TI - Aging and the lower urinary tract. AB - Lower urinary tract symptoms and disorders are prevalent and bothersome in the rapidly growing aging population. Common symptoms include urinary frequency, nocturia, urgency, and incontinence. Among older men, symptoms of voiding difficulty, such as hesitancy, slow stream, and straining, are also common. A variety of aging changes, as well as age-associated disease, predispose older people to the development of lower urinary tract symptoms. Medications used to treat conditions outside the lower urinary tract also can contribute to these symptoms. Physicians and other health professionals should routinely ask specific questions about lower urinary tract symptoms in their older patients and evaluate them thoroughly when they are present. PMID- 9332259 TI - Quality of life in geriatric patients with lower urinary tract dysfunction. AB - Lower urinary tract dysfunctions, such as urinary incontinence, detrusor instability, and benign prostatic hyperplasia, are prevalent in older adults. These conditions, which can occur alone or in combination, result in irritative or obstructive symptoms that can interfere with everyday functioning, leading to negative consequences on health-related quality of life. The nature and severity of these symptoms and the perception of their impact on daily activities can be quite variable. Until recently, relatively little was known about the effect of lower urinary tract dysfunctions on general health status and quality of life. An increasing research base is now available that shows the impact of different urologic dysfunctions in clinical and general populations. This article will provide a brief background on the definition and measurement of health-related quality of life and will summarize the literature about the quality of life of community-dwelling elderly patients with urinary incontinence or prostate conditions. Implications to guide clinical practice and future research will be derived. PMID- 9332260 TI - Postmenopausal vaginal atrophy and atrophic vaginitis. AB - Menopause is associated with a marked reduction in endogenous estrogen production. Lower levels of circulating blood estrogen have various deleterious effects, including those on the lower urinary tract. The vaginal epithelium becomes atrophied and dry, which can cause vaginal discomfort, itching, and dyspareunia. The epithelium may become inflamed and contribute to urinary symptoms, including frequency, urgency, dysuria, and incontinence. Diminished estrogen effects on periurethral tissues can contribute to pelvic laxity and stress incontinence. Changes in vaginal pH and normal flora may predispose older women to urinary tract infection. Although estrogen replacement therapy can result in maturation of the vaginal epithelium, the optimal form of administration and the dosage regimen for improving symptoms and quality of life of the geriatric female population have not been well studied. PMID- 9332261 TI - Nocturnal polyuria in the elderly person. AB - Aging often disturbs the normal circadian rhythm of urine production. The nocturia commonly seen with aging may result from the loss of nighttime vasopressin production or release that develops by childhood. Restoring the nocturnal increase in vasopressin can have a dramatic clinical response: improved quality of life and less risk of nighttime falls in carefully selected and accurately diagnosed patients. PMID- 9332262 TI - Benign prostatic hyperplasia. AB - Benign prostatic hyperplasia is a common and costly disease that affects older men. This article reviews the underlying histopathologic changes that cause benign prostatic hyperplasia. The symptoms and systematic evaluation of the patient are described, along with the current therapeutic options for treatment. PMID- 9332264 TI - Diagnostic assessment of geriatric urinary incontinence. AB - Urinary incontinence affects 15% to 30% of the population and 50% of those living in nursing homes. Care for incontinence is difficult because this condition is underreported by patients and underdiagnosed by physicians. This article describes the medical and nursing diagnostic assessment of urinary incontinence of geriatric populations, and the criteria for referral. Comprehensive review of urinary incontinence, including treatment, is available elsewhere. Although this article focuses on the assessment of urinary incontinence, the type of treatment being considered guides the scope of the evaluation, and therefore, treatment of incontinence is discussed. PMID- 9332263 TI - Urinary tract infection: an overview. AB - Urinary tract infection (UTI) remains very common. As many as 50% of women report having had at least one UTI in their lifetimes. Urinary tract infection is the most common cause of infection in nursing home residents and the most common source of bacteremia in the elderly population. Urinary tract infection occurs in patients with structurally or functionally abnormal urinary tracts (complicated UTI) and in patients with anatomically normal urinary tracts (uncomplicated UTI). Escherichia coli (E coli) is the most common cause of uncomplicated UTI, whereas antibiotic-resistant Enterobacteriaceae, enterococci, and Candida species often are the causes of complicated UTI. In this article we review current concepts of the epidemiology, microbiology, pathophysiology, clinical manifestations, diagnosis, and treatment of urinary tract infection. PMID- 9332265 TI - Behavioral interventions for incontinence in ambulatory geriatric patients. AB - Behavioral intervention is a group of therapies used to modify stress, urge, or mixed urinary incontinence by changing patient bladder habits or by teaching new skills. Three broad categories of behavioral treatment are reviewed: pelvic muscle exercise, biofeedback, and bladder training. The literature concerning each of these methods indicates that the treatments are effective for most community-dwelling older adults. More research is needed to identify the best methods for implementing these treatments, to explore the role of various components of treatment packages, to examine issues of durability, and to improve adherence to behavioral protocols necessary for long-term effectiveness. PMID- 9332266 TI - Pharmacologic treatment of lower urinary tract dysfunction in geriatric patients. AB - Medications to treat lower urinary tract dysfunction in older adults are selected to alter specific physiologic parameters. Pharmacotherapy alone results in modest clinical improvement. Because of the high prevalence of adverse drug reactions and polypharmacy in the geriatric population, medication should be used for those conditions that do not respond sufficiently to behavioral therapy. For stress incontinence, medications with alpha-adrenergic agonist properties are the mainstay of pharmacotherapy because they increase outlet resistance. Pharmacotherapy of urinary frequency and urge incontinence aims to decrease detrusor irritability and increase bladder capacity by inhibiting cholinergic stimulation of the bladder. In addition to these medications, in postmenopausal women, estrogen seems to have an additive effect for both urge and stress incontinence. More randomized, placebo-controlled, double-blinded clinical trials are needed that compare various pharmacologic agents and combinations, as well as pharmacotherapy with other forms of treatment for lower urinary tract dysfunction. PMID- 9332267 TI - Surgical treatment of urinary incontinence in geriatric women. AB - Surgical treatments have a limited role in the treatment of geriatric urinary incontinence. Patterns of problems exist with incontinence, including pelvic support defects and bowel and bladder dysfunction. Each of the major elements must be treated to achieve the best outcomes. Urodynamic testing should be used to confirm the cause of incontinence before selecting a surgical procedure. Minimally invasive procedures include periurethral collagen injections. PMID- 9332268 TI - Aortic root pseudoaneurysm after aortic dissection repair. PMID- 9332269 TI - Occult extraadrenal pheochromocytoma treated as diabetes mellitus. AB - Pheochromocytoma usually is associated with a combination of various manifestations caused by overproduction of catecholamines. We encountered a case of an occult, catecholamine-secreting pheochromocytoma. A 70-year-old man was admitted to the hospital because of anorexia. He had been treated for diabetes mellitus for 4 years; during this period he did not have any other symptoms related to pheochromocytoma. At admission, serum epinephrine, norepinephrine, and glucose levels and urinary excretion of total metanephrine were elevated. A tumor was detected in the left adrenal region and diagnosed as pheochromocytoma. After tumor resection, the increased levels of catecholamines and glucose and the decreased urinary C-peptide were normalized. This suggests that the pheochromocytoma caused hyperglycemia without other manifestations for a long time. PMID- 9332301 TI - Renewed interest in granulocyte transfusion therapy. PMID- 9332302 TI - The clinical significance of mutations of the P53 tumour suppressor gene in haematological malignancies. PMID- 9332303 TI - Acquired skewing of X-chromosome inactivation patterns in myeloid cells of the elderly suggests stochastic clonal loss with age. AB - More frequent skewing of X-chromosome inactivation patterns (XCIPs) occurs in the white blood cells of elderly females; this study was performed to determine whether this occurs in myeloid or lymphoid lineages. XCIPs were analysed in purified neutrophils and T cells from 80 females > 75 years and the results were compared with 23 cord blood and 94 younger adult blood samples. The degree of XCIP skewing in cord blood and younger adult blood cells was similar, with 3-4% having > 90% expression of one allele. Skewing was markedly increased in the neutrophils of elderly females, with 33% having > 90% expression of one allele (P < 0.0001). Extreme skewing was present in only 9% of the elderly T-cell samples and no evidence of T-cell clonality was found by PCR analysis of the TCR gamma gene. The high level of acquired skewing of the XCIPs in myeloid cells of the elderly suggests that with time there is a change in stem cell usage with stochastic loss of some of the original stem cells. This has major implications for the use of XCIP analysis in the diagnosis of myeloid malignancies in the elderly and for gene therapy into haemopoietic stem cells. PMID- 9332304 TI - Granulocyte-macrophage colony-stimulating factor abrogates transforming growth factor-beta 1-mediated cell cycle arrest by up-regulating cyclin D2/Cdk6. AB - The role of positive and negative cytokine interactions in G1 cell cycle regulation of haemopoietic cells was analysed by determination of the expression patterns of D-type cyclins and cyclin-dependent kinases (cdks) in SKM-1 myelodysplastic syndrome (MDS) cells incubated with granulocyte-macrophage colony stimulating factor (GM-CSF) and/or transforming growth factor-beta 1 (TGF-beta 1). TGF-beta 1 inhibited SKM-1 cell proliferation due to the cell cycle arrest in G1 phase. GM-CSF abrogated the TGF-beta 1-mediated G1 arrest in these cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that TGF-beta 1-mediated G1 arrest correlated with the down-regulation of cdk4, cdk6 and cyclin D2, and that abrogation of TGF-beta 1-mediated G1 arrest by GM-CSF correlated with the constitutive over-expression of cyclin D2 and cdk6 but not cdk4. These results suggest the importance of cyclin D2/cdk6 levels in abrogating G1 arrest in cells exposed to TGF-beta 1, and raise the possibility that the GM CSF-mediated up-regulatory pathway of signal transduction through cyclin D2/cdk6 differs from the TGF-beta 1-cdk4-mediated pathway in SKM-1 cells. This signal transduction pathway through cyclin D2/cdk6 might play an important role in haemopoietic regulation by the cytokine network. PMID- 9332305 TI - High-resolution cell division tracking demonstrates the FLt3-ligand-dependence of human marrow CD34+CD38- cell production in vitro. AB - Investigation of primitive human haemopoietic cell behaviour requires methodologies for monitoring asynchronously activated cells over several generations. We describe a high-resolution procedure for tracking 5- (and 6-) carboxyfluorescein diacetate succinimidyl ester (CFSE)- labelled human haemopoietic cells through six cell cycles based on the precise halving of their CFSE-fluorescence at each mitosis. Using this approach in combination with DNA or surface antigen staining, we show that the addition of Flt3-ligand (FL) to a cytokine cocktail consisting of Steel factor, IL-3, IL-6 and G-CSF increased the proportion of CD34+ (CD45RA/CD71)-, but not CD34+(CD45RA/CD71)+, human marrow cells initially recruited into division in vitro, shortened the overall cycle time of their progeny, and enhanced the production of a derivative CD34+CD38- population through several (up to four) cell generations. These studies also showed that during the first 4d there was no detectable apoptosis among the progeny of the CD34+(CD45RA/CD71)- cells generated in the presence of this four cytokine cocktail, regardless of the presence of FL. The availability of a technique for monitoring changes in the properties of individual cells as a function of their mitotic history and under conditions where they are asynchronously recruited to divide provides a new and powerful approach for studies of the regulation of primitive human haemopoietic cell proliferation and differentiation. PMID- 9332306 TI - Expression and functional analysis of two isoforms of the human GM-CSF receptor alpha chain in myeloid development and leukaemia. AB - The human granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) alpha chain RNA is alternatively spliced to yield receptor isoforms. Two of these, alpha 1 and alpha 2, differ in their cytoplasmic domains. Because the GM CSFR beta chain (beta c) is shared with the receptors for interleukins 3 and 5 it is possible that the alpha chain confers specificity on the GM-CSF response and that the different isoforms might refine this response further. Studies have been directed at determination of the respective biological roles of the alpha 1 and alpha 2 isoforms. Expression of the isoforms was examined by RNase protection analysis in normal granulocytes and a variety of cell lines of haemopoietic origin, at different stages of differentiation and activation. Expression was also analysed in cells from patients with a variety of leukaemic subtypes. Results demonstrated that the relative abundance of the isoforms was similar in all cell populations examined. The human GM-CSFR alpha 1 or alpha 2 receptors were independently expressed in the murine factor-dependent cell line FDC-P1, so that the properties of the receptors could be compared. Cell lines that expressed either receptor could be converted to growth in response to human GM-CSF and assumed a more differentiated phenotype when compared with the parental cell line. However, the morphology, expression of cell surface antigens and dose growth response characteristics did not differ significantly between cells that expressed either the alpha 1 or alpha 2 receptor. These studies demonstrate that the alpha 1 and alpha 2 subunits of the GM-CSF receptor are co-ordinately regulated in both normal and malignant haemopoiesis. Furthermore, each receptor is able to deliver both proliferative and differentiative signals to myeloid cells. PMID- 9332307 TI - An erythroid and megakaryocytic common precursor cell line (B1647) expressing both c-mpl and erythropoietin receptor (Epo-R) proliferates and modifies globin chain synthesis in response to megakaryocyte growth and development factor (MGDF) but not to erythropoietin (Epo). AB - A human megakaryocyte cell line (B1647) has been established from bone marrow cells obtained from a patient with acute myelogenous leukaemia (FAB M2). The cells were CD34-, CD33+, HLA-DR+, CD38+, and expressed the immunophenotypic markers of the megakaryocyte lineage (CD41 and von Willebrand factor). Moreover the cells expressed the c-mpl (thrombopoietin receptor) mRNA and protein. On the other hand, the B1647 cells also possessed erythroid lineage characteristics: the vast majority of cells were glycophorin positive, and about 10% of unstimulated cells stained with an anti-globin gamma chain MoAb. In addition, S1 protection analysis demonstrated expression of beta-globin mRNA, and Epo receptor (Epo-R) protein was detected by cytofluorimetric assay. Several growth factors, when tested alone or in combination, failed to influence the B1647 cell growth. A significant increase of cell proliferation was observed only after the addition, in serum-free culture, of recombinant human megakaryocyte growth development factor (MGDF), a recombinant c-mpl ligand encompassing the receptor-binding domain and identical to thrombopoietin (TPO), at concentrations ranging from 0.01 to 1 ng/ml. Interestingly, MGDF failed to induce megakaryocytic differentiation of the B1647 cells, but significantly increased the synthesis of the globin gamma chain. B1647 cells could be a useful model for studying the biological effect of TPO on common megakaryocyte and erythroid progenitors. PMID- 9332308 TI - Rat liver biliary epithelial cells support long-term production of haemopoietic progenitors from human CD34 cells. AB - In this study we report the supportive activity of rat liver epithelial cells (RLEC) on human haemopoiesis in the absence of exogeneously supplied growth factors. RLEC is a rat cell line derived from primitive biliary cells with epithelial characteristics which induce the long-term differentiation of hepatocytes through cell-cell contacts. We have established the ability of these cells to sustain long-term survival and multilineage differentiation of human haemopoietic progenitors from unfractionated bone marrow and growth-factor mobilized peripheral blood cells, and from human CD34+ and CD34+ CD38- haemopoietic cells, with a higher efficiency than the murine MS-5 stromal cell line: the numbers of committed progenitors recovered from RLEC cocultures after 8 weeks were 3-fold higher than from MS-5 cocultures, with an unusually high BFU-E production. Furthermore, using diffusible insert cultures, we demonstrated that, despite the lack of strong adhesive interaction between haemopoietic cells and RLEC, physical proximity was absolutely required for optimum stimulation of LTC IC by RLEC. Taken together, these results show that biliary epithelial cells support human haemopoiesis and cause speculation that common mechanisms might be used by RLEC to regulate both the hepatocyte and the haemopoietic progenitors differentiation. PMID- 9332309 TI - Bone marrow innervation regulates cellular retention in the murine haemopoietic system. AB - An anatomical analysis of the innervation of murine femora revealed intimate association of haemopoietic and stromal cells with nerve fibres. The mechanical denervation of these femora resulted in significant mobilization of cells into the peripheral blood within 24h. There was a decrease in femoral cellularity and analysis of the type of cells mobilized also revealed that there was an increase in progenitor cells in the peripheral blood. In non-splenectomized mice these progenitor cells were quickly cleared from the circulation. Chemical sympathectomy of mice with 6-hydroxydopamine resulted in decreased bone marrow cellularity without a change in bone marrow or peripheral blood progenitor cell numbers, nor the sustained rise in peripheral leucocytes observed with whole nerve denervation. These observations argue for selective control of mobilization by the nervous system and also indicate possible control of proliferation within the bone marrow. We conclude that the innervation has an important role in the maintenance of the blood-marrow interface, control of peripheral blood cell numbers, and mobilization of colony forming cells into the periphery. PMID- 9332310 TI - Two new mutations of the glucose-6-phosphate dehydrogenase (G6PD) gene associated with haemolytic anaemia: clinical, biochemical and molecular relationships. AB - In two unrelated Spanish males with glucose-6-phosphate dehydrogenase (G6PD) deficiency and haemolytic anaemia, and two different novel point mutations in the G6PD gene, have been identified. A C to T transition at nucleotide 406 resulting in a (136) Arg to Cys substitution and a C to G transition at nucleotide 1155 resulting in a (385) Cys to Trp substitution. These two molecular defects have not been described before and are designated G6PD Valladolid 406 C-->T and G6PD Madrid 1155 C-->G. In vitro biochemical characterization of both mutant enzymes showed important differences in their molecular properties according to their different clinical behaviour. In G6PD Valladolid, the mutation of which is located in exon 5, the normal in vitro heat stability may explain its mild clinical expression (low-grade haemolysis interrupted by an acute haemolytic crisis at age 70). In G6PD Madrid, the mutation, located in exon 10, results in a deficient variant associated with neonatal jaundice and life-long chronic nonspherocytic haemolytic anaemia (CNSHA). This finding further emphasizes the importance of this specific region of the G6PD gene in the stabilization of the G6PD molecule. Putative relationships between these single point mutations and the molecular properties of the mutant enzymes are also discussed. PMID- 9332312 TI - A murine model of human cold agglutinin disease. AB - Progress has been limited in the treatment of cold agglutinin (CA) disease by the absence of an animal model. We have recently studied at the molecular level one CA displaying the rare anti-Sia-1b specificity (CAGAS), CAGAS displays strong CA activity and is able to haemolyse mouse RBC in the presence of complement, thus constituting a suitable Ab for creating a murine model of CA disease. In the present work we introduced CAGAS VH and VL domains into eukaryotic expression vectors and transfected them into the non-secreting mouse myeloma X63 cell line. Clones expressing complete engineered pentameric IgM kappa CAGAS (eCAGAS) recapitulating the characteristics of serum CA (sCAGAS) could be obtained. The i.p. injection of eCAGAS to normal BALB/c mice induced a typical haemolytic anaemia, as demonstrated by the presence of spontaneous cold agglutination of RBC, induction of anaemia and significant reticulocytosis. Of interest, conspicuous bilateral ear loss was observed in one of these animals. In addition, i.p. injection of X63 transfected line into BALB/c nude mice induced ascites, typical haemolytic anaemia, and shortening of the mean RBC survival. These findings validate the practical interest of constructing a transgenic mouse model expressing eCAGAS. PMID- 9332311 TI - A potential determinant of enhanced crystallization of Hbc: spectroscopic and functional evidence of an alteration in the central cavity of oxyHbC. AB - The structural basis of the crystallizing tendencies of oxyHbC (beta 6Glu-->Lys), that produces haemolytic anaemia in homozygotes, is unknown. Using a fluorescent organic phosphate analogue (8-hydroxy-1,3,6-pyrenetrisulphonate), and conventional oxygen equilibrium studies, data suggest that the binding of inositolhexaphosphate (IHP) to oxyHbC differs from HbA, indicating perturbations of the oxyHbC central cavity, which was predicted from our earlier spectroscopic findings. To define the relationship between this conformational change in oxyHbC and its tendency to crystallize, the effect of four central cavity ligands on the crystallization rate was studied: a peptide containing 11 residues from the N terminal portion of band 3, the full cytoplasmic domain of band 3, 2,3 diphosphoglycerate and IHP. OxyHbC crystallization was accelerated by all these central cavity ligands and not by the appropriate controls. These central cavity changes become an excellent candidate for the dramatic increase in the crystallization rate of oxyHbC. PMID- 9332313 TI - Immune function in patients with beta thalassaemia receiving the orally active iron-chelating agent deferiprone. AB - Short-term deferiprone may reduce body iron in some patients with thalassaemia major. Concerns regarding potential immunosuppressive effects of deferiprone have been raised from results of animal studies and case reports in humans. We studied immune function in 57 thalassaemia patients: 36 treated with deferiprone (L1; CP020) and 21 treated with desferrioxamine (DFO). Circulating B lymphocytes were increased in all patient groups. No differences were detected between treatment groups in percentages of circulating lymphocytes, concentrations of IgG, IgM or IgA, specific antibody titres, complement levels, or in vitro lymphocyte proliferation. No clinically important infections were observed in any patient. These data suggest that no clinical or laboratory changes consistent with immuno suppression or immunodeficiency are observed during deferiprone therapy. PMID- 9332314 TI - An alternative to continuous subcutaneous infusion of desferrioxamine in thalassaemic patients. AB - Sixteen patients with thalassaemia major were treated with subcutaneous desferrioxamine (DF) 50 mg/kg/d, 5 consecutive days a week, for 8 weeks. Every other week the total dose was administered by 12 h infusion pump or by rapid injection of the same dose (25 x 2 mg/kg) twice a day. The two methods of DF administration produced no significant differences in urinary iron excretion. No significant changes in serum ferritin levels were observed at the end of the study. Compared with continuous infusion, rapid injection is equally efficacious, does not induce serious side-effects, is better accepted by the patients, and can improve their compliance to the iron-chelating therapy. PMID- 9332315 TI - Effects of in vivo administration of G-CSF on neutrophil and eosinophil adhesion. AB - The effect in vivo of G-CSF on neutrophil and eosinophil adhesion was studied after subcutaneous administration to six healthy individuals of human recombinant glycosylated G-CSF (lenograstim) (3 micrograms/kg) for 6 consecutive days. Basal adhesion and adhesion to E-selectin. VCAM-1 and ICAM-1 of neutrophil and eosinophil granulocytes were measured selectively. During G-CSF administration neutrophil basal adhesion increased from 7.4 +/- 3.9% (mean +/- SD) to 55.8 +/- 12.9% and 23.2 +/- 4.4%, 4 and 7 d, respectively, after start of the administration. At the same time points eosinophil basal adhesion increased from 7.1 +/- 2.4% to 37.7 +/- 6.1% and 13.1 +/- 5.3%, respectively. When adhesion was measured in the presence of Mn2+, which increases the functional activity of integrins, an even higher increase of neutrophil and eosinophil basal adhesion was noted 4 and 7 d, respectively, after start of G-CSF administration. In parallel with the enhanced basal adhesion neutrophil adhesion to E-selectin and ICAM-1 and eosinophil adhesion to E-selectin. VCAM-1 and ICAM-1 were significantly (P < 0.05) increased after 4 d of G-CSF administration as was neutrophil cell surface expression of CD11b and CD18. In vitro G-CSF induced minimal changes of granulocyte basal adhesion and inhibition of the adhesion to E selectin. 10 ng/ml TNF alpha significantly increased neutrophil and eosinophil basal adhesion and adhesion to VCAM-1 and ICAM-1. In summary, administration of G CSF to healthy subjects induced enhanced adhesion of neutrophil and eosinophil granulocytes, probably mediated by an increase of the functional capacity of beta 1- and beta 2-integrins. The induction of increased levels of TNF alpha might be one mechanism behind the in vivo effect of G-CSF administration. PMID- 9332316 TI - Activation of neutrophil function via CD66: differential effects upon beta 2 integrin mediated adhesion. AB - To further define the role of CD66 glycoproteins in the regulation of neutrophil function, we analysed the effects of a CD66 monoclonal antibody, Kat4c, which recognizes an epitope present on AB domains of CD66a, CD66b and CD66c. Intact Kat4c and F(ab')2 fragments were found to augment fMLP-induced oxidation of 1,2,3 dihydrorhodamine (oxidant species production) and beta 2 integrin-mediated adhesion to fibrinogen but did not promote beta 2 integrin-mediated binding of albumin coated latex beads. Since the latter assay is a sensitive indicator of neutrophil CD11b/CD18 functional activation, these results imply CD66 may exert differential effects upon beta 2 integrin activity. Neutrophil oxidant species production and spreading on fibrinogen substrates were further potentiated by cross-linking of Kat4c F(ab')2, in keeping with the suggestion that ligation of CD66 regulates neutrophil function. However, although intact Kat4c promoted beta 2 integrin-dependent homotypic neutrophil adhesion, F(ab')2 fragments were without effect, implying a role for Fc receptors in this effect which has previously been attributed to CD66. Together these data define more clearly the role of CD66 in regulation of neutrophil function and further suggest that augmented beta 2 integrin-mediated adhesion following CD66 ligation occurs independently of affinity regulation. PMID- 9332317 TI - Expression of adhesion molecules and functional stimulation in human neutrophils: modulation by GM-CSF and role of the Bcr gene. AB - Although devoid of proliferative capacity, polymorphonuclear neutrophils (PMN) express receptors for haemopoietic growth factors and need growth factors for survival and functional stimulation. This study showed that in vitro treatment of human PMN with GM-CSF for up to 48 h increases cell surface expression of the beta 2-integrin molecules CD11b/CD18 and CD11c/CD18 and of the receptor for the chemotactic peptide fMLP. Such modifications are usually expression of PMN activation. PMN treated with GM-CSF also displayed increased phagocytosis of latex particles and enhanced oxidative burst and superoxide anion release. Since integrins mediate PMN adhesion to endothelium, homotypic adhesion, chemotaxis/phagocytosis and the triggering of respiratory burst, our results suggested that functional stimulation of PMN persisted following prolonged exposure of PMN to growth factors and that it was not a temporary phenomenon which lasted only for the first 12-24 h of treatment. We also used oligonucleotides antisense to the Bcr gene mRNA to inhibit expression of the gene and evaluate its function in PMN, following the recent observation that PMN from Bcr-null mutant mice produced increased amounts of reactive oxygen metabolites upon activation. The antisense oligonucleotides had no effect on the parameters investigated. This may indicate that increased production of O2 by neutrophils in which the Bcr gene is not expressed requires either that gene expression is absent in the earlier stages of myeloid differentiation/maturation, so that when inhibition occurs in the terminally differentiated neutrophils their functional status is no longer influenced, or that the residual low-level expression of the gene which may be present in the antisense-treated cells is sufficient to provide a normal response to stimulation. PMID- 9332318 TI - Protein C and protein S in homozygous sickle cell disease: does hepatic dysfunction contribute to low levels? AB - The aim of this study was to confirm reports of low protein C (PC) and S (PS) concentrations in steady-state patients with homozygous sickle cell (SS) disease when compared to a racially matched normal haemoglobin (AA) control group and to examine the mechanisms of this reduction with respect to hepatic function, coagulation activation and haematological indices. In 36 SS patients and 35 AA race-matched controls PC (functional and immunoreactive), PS (free and total) were measured. C4B binding protein (C4B) was assessed by immunoelectrophoresis and D-dimer by ELISA. Hepatic function was assessed by prothrombin (PT) time (49 SS, 64 AA), factor V (34 SS, 36 AA) and factor VII concentrations (28 SS, 29 AA). Proteins induced in vitamin K absence or antagonism (PIVKA) were sought in 12 SS's. The relationship between PC, PS and total bilirubin, haemoglobin (Hb) F and reticulocyte count was also assessed. PC, PS and C4B were lower in SS disease. SS patients had longer PT times, and lower factor V and VII concentrations in comparison to AA controls. PC (functional and immuno-reactive) and free PS correlated with PT. Within SS genotype PT correlated negatively with factor V and factor VII. Factor V and VII were positively correlated. PIVKAs were not detected. There was no correlation between PC, PS and D-dimer, haemolytic rate or Hb F concentration. Prolongation of PT time, low factor V and VII suggest that hepatic dysfunction, rather than coagulation activation or haemolytic rate, accounts for the reduced concentrations of PC and PS in steady-state SS disease. The absence of PIVKAs suggests a hepatocellular problem. PMID- 9332319 TI - Fibrinogen Otago: a major alpha chain truncation associated with severe hypofibrinogenaemia and recurrent miscarriage. AB - A woman with a preliminary diagnosis of afibrinogenaemia was later found to have a functional fibrinogen of 0.06 mg/ml and markedly prolonged thrombin and reptilase times. The stoichiometry of fibrinopeptide release was normal but there was a gross delay in the polymerization of purified fibrin. Plasma protein electrophoresis showed an absence of normal fibrinogen and a novel anodal component which was confirmed as fibrinogen by immunofixation. Western blots of non-reducing SDS-PAGE gels indicated a molecular weight of 270 kD, compared to 340 kD for normal fibrinogen and similar analysis of reducing gels showed that the expected 67 kD A alpha chain was missing and replaced by a 30 kD band. This aberrant chain was not detected by the monoclonal antibody F-103, which recognizes the epitope formed by residues 259-276 of the A alpha chain. Cycle sequencing of the DNA encoding the F-103 epitope revealed the homozygous insertion of cytosine at position 4133 of the gene sequence. Predictably this translates as three new amino acids (268Gln-Glu-Pro) before termination at a new (TAG) stop codon. No abnormal A alpha chains could be detected in plasma from the woman's heterozygous son. The hypofibrinogenaemia observed is likely to be the result of diminished assembly and/or secretion of the truncated A alpha chains rather than enhanced extracellular degradation. PMID- 9332320 TI - A simplified statistical method for local INR using linear regression. European Concerted Action on Anticoagulation. AB - A simplified method of International Normalized Ratio (INR) derivation using linear regression of certified INR plotted against local prothrombin time (PT) results has been compared with INR from conventional orthogonal regression. Linear regression assumes error only with the local PT results whereas orthogonal regression assumes error with both reference and local results. The reliability of local INR derivation using lyophilized plasmas has been assessed in a collaborative study. INR from conventional fresh plasma International Sensitivity Index (ISI) calibrations have been compared with INR from calibrations with two types of lyophilized plasma, artificially depleted and coumarin. Although calibration slopes differed with the two types of analysis and the different lyophilized plasmas, both gave reasonable approximations to fresh plasma ISI calibrations. With orthogonal regression the overall percentage INR deviation was 5.25% with the artificially depleted plasmas and 6.85% for the results with lyophilized coumarins. With the linear regression, deviation was 8.40% for the artificially depleted plasmas and 5.05% for coumarin-treated patients' lyophilised-plasma. The simpler regression method appears to be worthy of further study as the present report has demonstrated that if the calibrant plasmas are accurately certified with the thromboplastin International Reference Plasma (IRP) results approximate to the conventionally determined INR using the manual PT technique. Coagulometers require further assessment. PMID- 9332321 TI - A sensitive and specific functional flow cytometric assay for the diagnosis of heparin-induced thrombocytopenia. AB - A functional flow cytometric assay (FCA) for the immediate diagnosis of heparin induced thrombocytopenia (HIT), with simultaneous compatibility testing for alternative anticoagulant therapies, has been developed to provide rapid and reliable results which effectively support patient management. The assay provides results within 1-2 h, uses readily available non-radioactive reagents, and employs standard equipment. Using the highly sensitive annexin V protein probe, the method detects activated platelets induced by heparin immune-complexes, with 300-fold increased binding to activated platelets. Twenty-five samples from patients clinically-suspected of having HIT (131 tests) and 10 normal control (NG) samples (36 tests) were simultaneously tested with unfractionated heparin (UH) and low-molecular-weight heparin (LMWH), and by the radioactive serotonin release assay (SRA) (62 and 16 tests respectively). The FCA highly correlated with the SRA, showing 100% specificity and 95% sensitivity. Moreover, the FCA exhibited higher resolution between positive and negative samples (an average value of 8.6-fold the NC versus 4.0-fold the NC by SRA). The LMWH showed concordant results with UH (r = 0.95). We conclude that the functional FCA for HIT is practical, specific and sensitive, thereby permitting the rapid diagnosis of HIT and the suitability of alternative therapies. PMID- 9332323 TI - The successful use of protein C concentrate during pregnancy in a patient with type 1 protein C deficiency, previous thrombosis and recurrent fetal loss. AB - The risks of venous thrombosis and fetal loss are increased in patients with protein C deficiency. We describe a patient with a history of thrombosis and recurrent fetal loss who was found to have type 1 protein C deficiency. In view of her history and intolerance of heparin preparations, she was treated with protein C concentrate during the first and third trimesters of her seventh pregnancy. The patient suffered no thromboembolic event during this pregnancy and gave birth to a healthy infant. The implications of the success of this therapeutic strategy are discussed. PMID- 9332322 TI - Danaparoid for cardiopulmonary bypass in patients with previous heparin-induced thrombocytopenia. AB - Anticoagulation for cardiopulmonary bypass in patients with heparin-induced thrombocytopenia requires the use of other anticoagulants. We report a case in whom this was achieved using the heparinoid danaparoid (Orgaran). Based on our experience and a review of the literature, we provide guidelines for managing these rare patients. A danaparoid dose substantially lower than that recommended by the manufacturer may minimize bleeding complications. PMID- 9332324 TI - Instability in the ATCT variable number tandem repeat locus VWF.VNTR I in intron 40 of von Willebrand factor gene. AB - The von Willebrand factor gene intron 40 variable number tandem repeat VWF.VNTR I exhibits 10 alleles making it highly polymorphic and useful for parentage and forensic testing, 45 unrelated families (210 meiotic events) were tested for VWF.VNTR I alleles. One spontaneous mutation was observed in a family member. Haplotype analysis demonstrated that this mutation was due to a gain of one motif repeat by a paternal allele. Sequence analysis confirmed the difference in the number of motif repeats between the proband and the alleles expressed by the parents. This instability emphasizes the importance of demonstrating exclusion in at least two separate loci in parentage testing. PMID- 9332325 TI - Bisphosphonates induce apoptosis in human myeloma cell lines: a novel anti-tumour activity. AB - Bisphosphonates are in widespread use to prevent bone resorption in a number of metabolic and tumour-induced bone diseases including multiple myeloma. Recent reports suggest that bisphosphonate treatment may be associated with an increase in patient survival, raising the possibility that these compounds may have a direct effect on the tumour cells. We have investigated whether the bisphosphonates clodronate, pamidronate and YM175 can directly affect the human myeloma cell lines U266-B1, JJN-3 and HS-Sultan in vitro. The effect of bisphosphonate treatment on cell number and cell cycle progression was examined using flow cytometry. The ability of bisphosphonates to induce apoptosis in human myeloma cell lines was determined on the basis of changes in nuclear morphology and of DNA fragmentation. Pamidronate and the more potent bisphosphonate. YM175, significantly decreased cell number (P < 0.001) in JJN-3 and HS-Sultan cells. YM175 also caused cells to arrest in the S-phase of the cell cycle in the JJN-3 cell line. Both pamidronate and YM175 also caused an increase in the proportion of cells with altered nuclear morphology (P < 0.05) and fragmented DNA, characteristic of apoptosis, in both JJN-3 and HS-Sultan cells. In contrast, clodronate had little effect on cell number and did not cause apoptosis at the concentrations examined. These data raise the possibility that some bisphosphonates could have direct anti-tumour effects on human myeloma cells in vivo. PMID- 9332326 TI - Inhibition of autophagy abrogates tumour necrosis factor alpha induced apoptosis in human T-lymphoblastic leukaemic cells. AB - The pattern and the sequence of tumour necrosis factor-alpha (TNF alpha) induced cell death in the acute T-lymphoblastic leukaemic cell line CCRF-CEM and its vinblastine-resistant subline CEM/VLB100 have been studied. Previously, we found that the CEM/VLB100 cell line was more sensitive to TNF alpha-induced killing than its parental CCRF-CEM cell line. TNF alpha-induced cell death showed an apoptotic pattern, as detected by agarose electrophoresis, flow cytometry and transmission electron microscopy (TEM). TEM images revealed that autophagy and codensed mitochondria occurred earlier than nuclear fragmentation. The specific inhibitor of autophagy, 3-methyladenine (3MA), inhibited the formation of autophagosomes. TNF alpha-induced DNA fragmentation and cytolysis were completely inhibited by 10 mM 3MA. Inhibition of the fusion of lysosomes with autophagosomes by asparagine did not block TNF alpha-induced apoptosis. In addition, amino acid and protein deprivation enhanced TNF alpha-induced autophagy but not apoptosis. We propose that the early stages of autophagy are required for, but do not necessarily result in, TNF alpha-induced apoptosis. PMID- 9332327 TI - Mitochondrial electron transport chain activity, but not ATP synthesis, is required for drug-induced apoptosis in human leukaemic cells: a possible novel mechanism of regulating drug resistance. AB - There is increasing evidence for an association between mitochondrial function and susceptibility to apoptosis. It has been shown that the vinblastine-resistant leukaemic cell line CEM/VLB100 has a more active mitochondrial electron transport chain (ETC) than the parental CCRF-CEM cell line. Inhibition of mitochondrial DNA replication by ethidium bromide (EB) depleted the activity of the ETC and reduced cellular respiratory rate. Depletion of mitochondrial DNA was associated with increased resistance to vinblastine-induced apoptosis in both cell lines. In contrast, the highly specific inhibitor of the energy producing mitochondrial enzyme F1Fzero-ATPase, oligomycin, rendered CEM/VLB100 cells more sensitive to vinblastine by inhibiting the energy-dependent P-glycoprotein (Pgp) pump, suggesting that the effect of EB is independent of energy generation and ATPase activity. Both mitochondrial ETC depletion and ATPase inhibition decreased vinblastine-induced cell cycle changes in the CCRF-CEM cell line, suggesting that cell cycle changes are dependent on ATP generation. However, EB-induced ETC depletion in CEM/VLB100 cells inhibited apoptosis in response to high concentration of vinblastine, but not G2M arrest. We suggest that: (1) over expression of Pgp by drug-resistant cells may up-regulate mitochondrial energy production; (2) mitochondrial ETC activity is required for DNA fragmentation in response to vinblastine, but the mechanism is independent of Pgp activity and ATP generation; (3) down-regulation of mitochondrial ETC activity may confer resistance to vinblastine-induced apoptosis; (4) the mitochondrial ETC is involved in vinblastine-induced apoptosis downstream of microtubule disruption and cell cycle changes. PMID- 9332328 TI - Interferon-gamma induced proliferation of human myeloid leukaemia cell lines. AB - Interferon-gamma (IFN-gamma) is a pleiotropic cytokine involved in the regulation of various phases of immune and inflammatory responses; it also has anti-viral and anti-proliferative activity. Using continuous human leukaemia cell lines as model systems, we found that IFN-gamma stimulated the proliferation of leukaemic myeloid cells; this effect was specifically neutralized by an anti-IFN-gamma monoclonal antibody (McAb). No proliferative response was seen in autonomously growing cell lines; however, 11/19 constitutively growth factor-dependent cell lines showed a significant response in short-term proliferation assays upon incubation with IFN-gamma. The stimulation indices ranged from 2 to 37 compared with the untreated control cells; the EC50 values for these cell lines were in the range of 0.1-0.6 ng/ml IFN-gamma. Flow cytometric analysis demonstrated heterogeneity in the expression of the IFN-gamma receptor, as it was found on 37 97% of the cells per cell line. The effects of IFN-gamma on proliferation triggered by a spectrum of 10 other cytokines were variable, and both stimulation and attenuation of the proliferative responses were seen in different cell lines. Under serum-free culture conditions, IFN-gamma acted as a survival factor suppressing apoptosis. As has been described for other functional processes triggered by IFN-gamma, the proliferation-inducing activity of IFN-gamma also led to activation of the signal transducing element STAT 1. Thus, IFN-gamma can induce myeloid leukaemia cells to proliferate and can modulate their proliferative response to other cytokines. Therefore IFN-gamma may be a pathologically relevant ligand for leukaemic cell proliferation in vivo. In physiological settings, IFN-gamma might be a bifunctional regulator of haemopoietic cell proliferation, depending on other differential co-signals from the micro-environment. PMID- 9332329 TI - A randomized comparison of liposomal versus conventional amphotericin B for the treatment of pyrexia of unknown origin in neutropenic patients. AB - One hundred and thirty-four adults and 204 children were randomized in two prospective, parallel comparative multicentre trials to receive either conventional amphotericin B 1 mg/kg/d (c-AMB), liposomal amphotericin B 1 mg/kg/d(L-AMB1) or liposomal amphotericin B 3 mg/ kg/d (L-AMB3). Patients were entered if they had a pyrexia of unknown origin (PUO) defined as temperature of 38 degrees C or more, not responding to 96 h of systemic broad-spectrum antibiotic treatment, and neutropenia (< 0.5 x 10(9)/l). The safety and toxicity of liposomal amphotericin B was compared with that of conventional amphotericin B. Efficacy of treatment was assessed, with success defined as resolution of fever for 3 consecutive days (< 38 degrees C) without the development of any new fungal infection. Clinical and laboratory parameters were collected for safety analysis. In both the paediatric and adult populations, L-AMB treated patients had a 2-6-fold decrease in the incidence (P < or = 0.01) of test-drug-related side-effects, compared to c-AMB. Severe trial-drug-related side-effects were seen in 1% of L-AMB treated patients, in contrast to 12% of patients on c-AMB (P < 0.01). Nephrotoxicity, in the patient subset not receiving concomitant nephrotoxic agents, defined as a doubling from the patients baseline serum creatinine level, was not observed in the L-AMB1 arm whereas the incidence was 3% in patients on L-AMB3 and 23% in those on c-AMB (P < 0.01). Moreover, time to develop nephrotoxicity was longer in both L-AMB arms than c-AMB (P < 0.01). Severe hypokalaemia was observed less frequently in both L-AMB arms (P < 0.01). Analysis was by intention-to-treat and included all patients randomized. Success was defined by a minimum of 3 consecutive days with fever (< 38 degrees C) continuing to study end indicated by recovery of neutrophils to 0.5 x 10(9)/l. Addition of systemic antifungal therapy or development of systemic fungal infection were failures as was persistent fever to study end. Efficacy assessments indicated success in 49% of the total group treated with c-AMB, 58% of patients responded to L-AMB1 and 64% to L-AMB3. A statistically significant difference was found between c-AMB and L-AMB3 (P = 0.03) but a Kaplan-Meier analysis of time to differvescence of fever showed there was no significant difference between the arms. It was concluded that liposomal amphotericin at either 1 or 3 mg/kg/d was significantly safer than conventional amphotericin B in children and adults. The main aim of this open-label study was to compare safety between the three trial arms. However, we provide evidence for an equivalent or possibly superior efficacy of liposomal amphotericin with regard to resolution of fever of unknown origin. Subsequent trials should compare amphotericin preparations in defined fungal infections. PMID- 9332330 TI - BCL6 gene rearrangements also occur in marginal zone B-cell lymphoma. AB - Marginal zone B-cell lymphoma (MZBCL) represents a distinct subtype of B-cell non Hodgkin's lymphoma (NHL) which has been recently recognized and defined as a disease entity. Cytogenetically, these lymphomas reveal a high prevalence of trisomy 3, and recent data obtained by comparative genomic hybridization indicate that the chromosomal regions 3q21-23 and 3q25-29 might be of particular pathogenetic significance. We identified structural chromosomal abnormalities involving the region 3q27 and rearrangements of the BCL6 proto-oncogene in three out of 34 (9%) well-defined cases of extranodal, nodal and splenic MZBCL using cytogenetic analysis. Southern blot, and fluorescence in situ hybridization (FISH). All three cases were characterized by a t(3;14)(q27;q32). Two of them showed additional chromosomal abnormalities including trisomy 3, which was found in one case. The patients displayed extranodal disease and did not demonstrate any striking clinical and histological differences when compared with MZBCL lacking BCL6 rearrangement. The present study for the first time demonstrates the occurrence of t(3;14)/BCL6 gene rearrangement in MZBCL, thus suggesting a role of the BCL6 proto-oncogene in the pathogenesis of MZBCL. PMID- 9332331 TI - Genetic heterogeneity of AIDS-related small non-cleaved cell lymphoma. AB - AIDS-related small noncleaved cell lymphoma (AIDS-SNCCL) includes Burkitt's lymphoma (BL) and high-grade B-cell Burkitt-like lymphoma (BLL). Due to the marked polymorphism of AIDS-related non-Hodgkin's lymphomas (AIDS-NHL), the morphologic distinction between these two types of lymphomas is frequently controversial, although it may bear clinical relevance. Although the molecular features of AIDS-BL have been clarified to a certain extent, the genetic peculiarities of AIDS-BLL have not been investigated in detail. In this study we have compared morphologic and genetic features of AIDS-BL and AIDS-BLL in a blind coded fashion. Molecular studies were focused on the genetic lesions known to be implicated in AIDS-NHL, including alterations of c-MYC, BCL-6, p53, deletions of 6q, as well as infection by EBV and HHV-8. Alterations of c-MYC occurred in 10/10 AIDS-BL, whereas they were restricted to 2/10 AIDS-BLL (P < 0.01). Mutations of p53 were present in 5/10 AIDS-BL, whereas they were consistently absent among AIDS-BLL (n = 10; P < 0.05). Infection by EBV occurred in 30% of both AIDS-BL and AIDS-BLL. Rearrangements of BCL-6, deletions of 6q and infection by HHV-8 scored consistently negative in both AIDS-BL and AIDS-BLL. Based on the genetic lesions tested, the molecular profile of AIDS-BLL appears to be closer to that of AIDS related diffuse large cell lymphoma (AIDS-DLCL) than to that of AIDS-BL. In contrast to AIDS-BLL however, AIDS-DLCL carried rearrangements of BCL-6 in a fraction of cases (2/9). This study, the largest of its kind reported so far, suggests that AIDS-BL and AIDS-BLL have a different molecular pathogenesis and that characterization of genetic lesions may help to distinguish between these two lymphomas. PMID- 9332332 TI - t(8;20)(q22;p13): a novel variant translocation of t(8;21) in acute myeloblastic leukaemia. AB - A novel variant translocation t(8;20)(q22;p13) detected by karyotype analysis of bone marrow cells using R- and G-banding techniques, is reported in a case of M2 acute myeloid leukaemia (AML). The leukaemic cells were indistinguishable morphologically from that of M2-AML with t(8;21)translocation. RT-PCR revealed no AML1/ETO fusion transcript, but the wild-type ETO-3' was expressed in the bone marrow cells suggesting that t(8;20) is a true simple variant translocation of t(8;21), and that a fusion gene consisting of ETO and an unidentified gene located in band 20p13 may exist in our case. Further study is required to clarify the entity of the assumed fusion gene. PMID- 9332333 TI - Factors influencing collection of peripheral blood progenitor cells following high-dose cyclophosphamide and granulocyte colony-stimulating factor in patients with multiple myeloma. AB - We treated 103 multiple myeloma (MM) patients with 7 g/m2 cyclophosphamide (Cy) followed by 300 micrograms G-CSF/d to harvest peripheral blood progenitor cells (PBPC). PBPC autografts containing > 2.0 x 10(6) CD34+ cells per kg body weight were obtained at the first attempt from 90/100 evaluable patients. The most significant factor predicting impairment of PBPC collection was the duration of previous melphalan treatment (P < 0.0001). In multivariate discriminate analysis, treatment with melphalan during the most recent chemotherapy cycles prior to mobilization (P = 0.0727) and previous radiotherapy (P = 0.0628) had a marginally significant negative influence on the efficacy of PBPC collection. We found no reduced functional capacity of CD34+ cells to restore haemopoiesis after myeloablative treatment related to the duration of melphalan exposure. At the time of best response to conventional treatment, a median paraprotein reduction of 21% was achieved following high-dose cyclophosphamide (HD-Cy). Two heavily pretreated patients died and one patient developed pulmonary toxicity W.H.O. grade IV following HD-Cy. Potential transplant candidates should undergo mobilization and harvesting of PBPC before melphalan-containing treatment. Combinations of haemopoietic growth factors and their dose-modifications should be investigated to improve PBPC collection, to allow a dosage reduction of the mobilization chemotherapy. PMID- 9332334 TI - Analysis of the effect of prior therapy on progenitor cell yield: use of a chemotherapy scoring system. AB - A quantitative analysis of peripheral blood stem cell (PBSC) yield, measuring absolute numbers of CD34+ cells x 10(6)/kg and CFU-C x 10(4)/kg was performed in 74 consecutive patients. The interval or 'gap' from the end of previous chemotherapy to the date of priming was recorded in weeks. Geometric mean CD34 and CFU-C values were significantly higher in patients with a score of < or = 60 compared to those with score > 60 (P = 0.003 and 0.02, respectively) and a significant difference in CD34 values was also found when scores of < or = 38 were compared with scores > 38 (P = 0.003), with the difference in CFU-C values approaching significance (P = 0.08). Patients exposed to toxicity factor 4 drugs had significant lowering of both CD34 and CFU-C values (P < 0.001 and P = 0.038) and this emerged as the only independent factor when analysed using linear regression. No significant difference in the geometric mean CD34 or CFU-C values of patients was found in any of the gap categories analysed. Prior exposure to toxicity factor 4 drugs had a significant adverse effect on stem cell yield and should be avoided or minimized prior to stem cell harvesting. PMID- 9332335 TI - Outcome of transplantation for standard-risk leukaemia with grafts depleted of lymphocytes after conditioning with an intensified regimen. AB - One hundred and eighty-one consecutive patients with standard-risk leukaemia were transplanted with HLA-identical sibling grafts depleted of lymphocytes using counter-flow centrifugation. In 116 patients, standard conditioning was intensified by the addition of anthracyclines. Multivariate analysis revealed significantly more acute GVHD > or = grade 2 and a trend towards more chronic GVHD in patients conditioned with the addition of anthracyclines. For all patients the risk for chronic GVHD, but not for acute GVHD increased with a higher number of T cells in the graft. The projected 5-year probability of relapse was significantly lower in the group of patients conditioned with anthracyclines; 26% versus 52% (P = 0.015). In multivariate analysis the addition of anthracyclines to the conditioning regimen was the only significant factor contributing to a lower probability of relapse. The projected 5-year probability of leukaemia-free survival [LFS] in the patients conditioned with and without the addition of anthracyclines was 56% and 36%, respectively (P = 0.004). In multivariate analysis the addition of anthracyclines to the conditioning regimen correlated significantly with a lower number of mixed chimaeras in patients at 6 and 12 months after BMT. Mixed chimaerism at 6 months after transplantation did not significantly correlate with a higher incidence of relapse in further follow up. In contrast, mixed chimaerism at 12 months after BMT was significantly associated with higher relapse rate. We conclude that the addition of anthracyclines to the conditioning regimen improves outcome of BMT using T-cell depleted grafts. PMID- 9332336 TI - Transplantation of HLA-mismatched CD34+ selected cells in patients with advanced malignancies: severe immunodeficiency and related complications. AB - This trial was designed to test the use of CD34+ selected haemopoietic stem cells (HSC) in HLA-mismatched donor-recipient pairs, following intensive conditioning with thiotepa, antilymphocyte globulin (ALG), cyclophosphamide and single-dose total-body irradiation (sTBI). 10 patients aged 16-50 with advanced malignancies and a two- or three-antigen mismatched family donor entered this study. Donor marrow and G-CSF primed peripheral blood cells were processed separately on CD34 columns (Ceprate). The median number of infused CD34+ cells were 5.66 x 10(6)/ kg, with 0.55 x 10(6)/kg CD3+ cells. Nine patients received cyclosporin for graft versus-host disease (GvHD) prophylaxis. Median neutrophil counts on day 21 were 2 x 10(9)/l with a median platelet count of 60 x 10(9)/l, but CD4 counts remained extremely depressed throughout the study. Acute GvHD was scored as grade 0-I in two patients, as grade II in seven, and grade III in one. Eight patients died at a median interval of 72 d from HSCT (range 20-144) due to cytomegalovirus (CMV) associated interstitial pneumonitis (IP) (n = 5), renal failure (n = 1). GvHD (n = 1) and Aspergillus meningitis (n = 1). Two patients are alive 365-495 d post transplant, one in remission and one in relapse. This study suggests that large numbers of positively selected mismatched HSC can rapidly engraft after intensive conditioning regimen: however, profound post-transplant immunodeficiency leads to a high risk of lethal infectious complications. PMID- 9332337 TI - Detection of maternal cells in human fetal blood during the third trimester of pregnancy using allele-specific PCR amplification. AB - Using a highly sensitive allele-specific PCR amplification method, we have previously shown that maternal cells could be detected in all 10 cord bloods tested. This raised the question of whether maternal cells are released into cord blood during the process of delivery or whether they are already present during pregnancy. We have now used the same PCR method to detect the presence of maternal cells in nine fetal blood samples collected at different gestational ages. Maternal cells were detected in eight samples obtained between 24 and 35 weeks of gestation. They were estimated to amount between 10(-4) and 10(-5) of nucleated fetal blood cells. In two cases mononuclear and polymorphonuclear cell fractions were separated by Ficoll gradient centrifugation and maternal cells were detected as comparable levels in both fractions. Maternal cells could not be detected in the one fetal blood sample obtained at 20 weeks of gestation, suggesting that maternal cells could appear at detectable levels in fetal blood during the third trimester of pregnancy. These results are discussed in terms of materno-fetal immune tolerance and of transmission of viruses (and more specifically of the human immunodeficiency virus) from mother to child. PMID- 9332338 TI - Successful second bone marrow transplant for Fanconi's anaemia following escalation of conditioning. AB - Allogeneic bone marrow transplantation represents the treatment of choice for severe bone marrow failure in patients with Fanconi's anaemia (FA). In view of the increased sensitivity to alkylating agents documented in this condition, much attention has focused on reducing the conditioning chemotherapy. We present a 13 year-old girl in whom sibling allogeneic BMT after conditioning with low-dose cyclophosphamide only resulted in graft rejection. However, a second transplant using the same donor proved successful following a more intensive conditioning regimen. This case demonstrates the phenotypic variability of FA, and highlights the need for tailoring the conditioning regimen for a given patient. PMID- 9332339 TI - Endothelial cell precursors are normal components of human umbilical cord blood. AB - Endothelial cells are part of the normal bone marrow stroma. We have previously shown human umbilical cord blood (UCB) does not produce stroma in standard long term cultures. Highly enriched (93-98%) UCB CD34+ cells were cultured for 6 weeks with interleukin-2 and conditioned medium from the 5637 carcinoma cell line (n = 4). The resulting 'fibroblast like' cells were shown to be endothelial by expression of von Willebrand factor (VWF), ICAM-1 (CD54), E-selectin (CD62E) and PECAM (CD31). Endothelial monolayers seeded with CD34+ UCB cells supported expansion of colony forming cells and CD34+ cells. We conclude that endothelial cell precursors circulate in UCB, and may be derived from the CD34+ cell fraction. PMID- 9332340 TI - Could HGV infection be implicated in lymphomagenesis? PMID- 9332341 TI - CD34 epitope expression on haemopoietic tissues. PMID- 9332342 TI - Rare combination of homozygous sickle cell disease (Hb SS) and haemophilia B in a paediatric patient. PMID- 9332343 TI - Refrigerated stored specimens are not reliable for delayed INR determination. PMID- 9332344 TI - Isolation of a cDNA encoding a novel subtilisin-like protease (Pr1B) from the entomopathogenic fungus, Metarhizium anisopliae using differential display-RT PCR. AB - Reverse transcription differential display PCR (RT-DD-PCR) was used to identify genes that are specifically expressed by Metarhizium anisopliae when it contacts the host insect cuticle. Using a homology-based subtilisin-like protease primer we identified a hitherto unsuspected differentially expressed subtilisin-like protease (Pr1B) encoding gene. The deduced amino acid sequence shows 54% similarity to the well characterized Pr1A subtilisin of M. anisopliae and karyotype analysis revealed that Pr1A and Pr1B are located on separate chromosomes. Like Pr1A, Pr1B is synthesized as a large precursor (1158 nucleotides; deduced molecular mass = 40031 Da) containing a signal peptide, a propeptide and the mature protease (283 aa; deduced molecular mass = 28714 Da). However, Pr1B possesses several substitutions in the highly conserved sequences comprising the active sites of subtilisins. In particular, the substitution of Thr220 by serine is unique to Pr1B. Substitution of Asn155 by glycine is also very unusual, and we discuss the likely effects these changes will have on the catalytic efficiency of Pr1B. PMID- 9332345 TI - Paralogous histidine biosynthetic genes: evolutionary analysis of the Saccharomyces cerevisiae HIS6 and HIS7 genes. AB - The HIS6 gene from Saccharomyces cerevisiae strain YNN282 is able to complement both the S. cerevisiae his6 and the Escherichia coli hisA mutations. The cloning and the nucleotide sequence indicated that this gene encodes a putative phosphoribosyl-5-amino-1-phosphoribosyl-4-imidazolecarboxiamide isomerase (5' Pro FAR isomerase, EC 5.3.1.16) of 261 amino acids, with a molecular weight of 29,554. The HIS6 gene product shares a significant degree of sequence similarity with the prokaryotic HisA proteins and HisF proteins, and with the C-terminal domain of the S. cerevisiae HIS7 protein (homologous to HisF), indicating that the yeast HIS6 and HIS7 genes are paralogous. Moreover, the HIS6 gene is organized into two homologous modules half the size of the entire gene, typical of all the known prokaryotic hisA and hisF genes. The structure of the yeast HIS6 gene supports the two-step evolutionary model suggested by Fani et al. (J. Mol. Evol. 1994; 38: 489-495) to explain the present-day hisA and hisF genes. According to this idea, the hisF gene originated from the duplication of an ancestral hisA gene which, in turn, was the result of an earlier gene elongation event involving an ancestral module half the size of the extant gene. Results reported in this paper also suggest that these two successive paralogous gene duplications took probably place in the early steps of molecular evolution of the histidine pathway, well before the diversification of the three domains, and that this pathway was one of the metabolic activities of the last common ancestor. The molecular evolution of the yeast HIS6 and HIS7 genes is also discussed. PMID- 9332346 TI - The 5'-upstream region of the rat phospholipase C-beta 3 gene contains two critical Sp1 sites and an HIV Inr-like element. AB - The 5'-upstream region of the rat phospholipase C-beta 3 gene (PLC-beta 3) has been cloned and characterized. Sequence analysis of the 5'-upstream region showed that it contains a GC-rich region (-166 to +1: 79%) and multiple binding sites for the transcription factors Sp1, AP-1 and AP-2, but does not contain a canonical TATA box. Primer extension analysis of total RNA isolated from rat glial cell C6Bul revealed that single transcription start point (tsp) is located at an initiator (Inr) element similar to that found in the HIV promoter. Gel mobility shift and competitive mobility shift assays indicated that this Inr element forms a DNA-protein complex with the HIV Inr-binding protein, LBP-1/CP2 or a homologue. In order to localize functional elements of the 5'-upstream region of the rat PLC-beta 3 gene, 5'-deletion fragments were cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. Transient transfection analyses of the 5'-deletion mutants identified a crucial promoter element located at -128 to -14. Supershift mobility assays, site-directed mutagenesis and DNase I footprints indicated that Sp1 binds to three GC boxes within the sequence between -128 and -14 of the PLC-beta 3 promoter. Transient transfection analyses of promoter constructs containing site-specific mutation(s) of these three GC boxes demonstrated that two GC boxes, located proximal to the tsp, are important elements for normal promoter activity. PMID- 9332347 TI - Cloning and characterization of a copy of Tirant transposable element in Drosophila melanogaster. AB - A Tirant element, inserted at the 5' end of the mitochondrial glutamine synthetase (mt-gs) gene in a mutant allele giving rise to a recessive female sterility phenotype, was cloned and utilized to characterize this novel retrotransposable element of the Drosophila melanogaster genome. The 5.3 kb element present in the fs(2) PM11-19 mt-gs allele possesses a 417 bp long terminal repeat (LTR) at both ends. There is a serine tRNA binding site downstream of the 5' LTR sequence and a polypurine tract upstream of the 3' LTR end. The insertion leads to the duplication of a host-site CGCG sequence. In situ hybridization to salivary glands chromosomes showed evidence of the mobile nature of the element. The DNA sequencing of the cloned 5.3 kb element revealed that Tirant possesses an open reading frame (ORF) that shows similarity with the envelope protein encoded by the gypsy and 297 retrotransposons. In addition, the cloned element appears to be a subgenomic fragment of a not yet identified complete element, because only the integrase domain of the reverse transcriptase gene is found. PMID- 9332348 TI - Cloning and characterization of a new alpha-amylase gene from Streptomyces lividans TK24. AB - Streptomyces lividans TK24 possesses a very weak amylolytic activity, nevertheless Southern blot analysis carried out at high stringency revealed that this strain does contain a gene strongly related to the well expressed alpha amylase gene (amlSL) of Streptomyces limosus. To clone this related gene, three genomic banks of S. lividans TK24 were constructed into the multicopy plasmid vector pIJ699 and transformed into the same strain. Two different genes were isolated. One (amlA) has been previously described, whereas the other (amlB) has never been described. Sub-cloning experiments localized amlB to a 3 kb BamHI-NotI fragment that was sequenced. Frame analysis on sequence data revealed the presence of a 1719 bp long open reading frame encoding a 573 amino acid protein of 61214 kDa. Northern blot analysis identified a unique 1.8 kb monocistronic transcript. Primer extension allowed the localization of the transcription start point 108 bp upstream of the translational start codon and demonstrated that the gene was transcribed from a unique typical eubacterial-like promoter. AmlB shares 74.7% amino acid identity with the alpha-amylase of S. limosus and only 27.2% with the amylolytic enzyme encoded by amlA. PMID- 9332349 TI - Identification and transcriptional analysis of a Treponema pallidum operon encoding a putative ABC transport system, an iron-activated repressor protein homolog, and a glycolytic pathway enzyme homolog. AB - We have characterized a 5.2-kilobase (kb) putative transport related operon (tro) locus of Treponema pallidum subsp. pallidum (Nichols strain) (Tp) encoding six proteins: TroA, TroB, TroC, TroD, TroR and Phosphoglycerate mutase (Pgm). Four of these gene products (TroA-TroD) are homologous to members of the ATP-Binding Cassette (ABC) superfamily of bacterial transport proteins. TroA (previously identified as Tromp1) has significant sequence similarity to a family of Gram negative periplasmic substrate-binding proteins and to a family of streptococcal proteins that may have dual roles as substrate binding proteins and adhesins. TroB is homologous to the ATP-binding protein component, whereas TroC and TroD are related to the hydrophobic membrane protein components of ABC transport systems. TroR is similar to Gram-positive iron-activated repressor proteins (DesR, DtxR, IdeR, and SirR). The last open reading frame (ORF) of the tro operon encodes a protein that is highly homologous to the glycolytic pathway enzyme, Pgm. Primer extension results demonstrated that the tro operon is transcribed from a sigma 70-type promoter element. Northern analysis and reverse transcriptase-polymerase chain reactions provided evidence for the presence of a primary 1-kb troA transcript and a secondary, less abundant, troA-pgm transcript. The tro operon is flanked by a Holliday structure DNA helicase homolog (upstream) and two ORFs representing a purine nucleoside phosphorylase homolog and tpp15, a previously characterized gene encoding a membrane lipoprotein (downstream). The presence of a complex operon containing a putative ABC transport system and a DtxR homolog indicates a possible linkage between transport and gene regulation in Tp. PMID- 9332350 TI - XLS13A and XLS13B: SRY-related genes of Xenopus laevis. AB - SRY-related cDNAs, XLS13A and XLS13B, have been isolated from Xenopus laevis ovary. The cDNAs encode polypeptides of 382 and 375 amino acids, respectively. Nucleotide sequences of the two cDNAs are highly homologous to each other. The type-A and type-B XLS13 proteins, and xSox13 reported previously share an identical high mobility group (HMG) box at the amino acid level, although they contain silent nucleotide alterations. The HMG box exhibits strong similarity (> 93% amino acid identity) to those of mouse Sox4/human SOX4 and chicken Sox11/human SOX11. The size of XLS13A/XLS13B mRNA was estimated to be 2.8 knt in Xenopus ovary by Northern analysis. Reverse transcription/polymerase chain reaction (RT/PCR) assay indicated that XLS13A and XLS13B mRNAs are present in various tissues of adult frog. The mRNAs of XLS13A and XLS13B of maternal origin found in unfertilized eggs disappear in the early stages of the Xenopus embryo. DNA-binding properties of the XLS13 HMG domain were examined by electrophoretic mobility shift assay (EMSA). The HMG domain preferentially binds to the canonical target sequence of SOX proteins, AACAAT, in vitro. PMID- 9332351 TI - CASP, a novel, highly conserved alternative-splicing product of the CDP/cut/cux gene, lacks cut-repeat and homeo DNA-binding domains, and interacts with full length CDP in vitro. AB - Human CDP/cut and its murine counterpart, cux1/CDP are homeodomain repressor proteins in the family of Drosophila Cut. Northern blot analysis reveals complex alternative splicing, including forms too small to encode the full 1505 amino acid protein. We have characterized a CDP/cut alternatively spliced cDNA (CASP) of 3.4 kb. Human CASP, a predicted 678 amino acid polypeptide, shares 400 amino acids with CDP, but has an alternate N terminal exon of 20 aa, and the C-terminal 258 amino acids diverge from CDP/cut entirely. As the unique C-terminus of CASP lacks the three 'cut-repeats' and homeodomain of CDP/cut, we predict it does not bind DNA. Murine CASP, 96% similar to human, shares these features. Database searches identify homologs in chicken (86% identical to human CASP) and yeast (29% identical to human). Murine CASP mRNA is ubiquitous in mouse tissues and in tissue-culture cell lines. We generated a specific antiserum against the unique C terminus of CASP, and used this reagent to demonstrate that CASP protein is expressed as an approx. 80 kDa protein in human and murine cells. Co-translation of in vitro-translated CDP and CASP mRNA, followed by immunoprecipitation with specific anti-CASP IgG, shows that CASP polypeptide can from a complex with CDP. Studies of the intron/exon structure of the murine cux/CDP/mCASP locus (>> 100 kb) reveal that the unique 3' exons of CASP are interposed between cut-repeats 2 and 3 of the cux gene. We speculate that a primordial CASP-like gene captured a cut-repeat-homeobox gene to give rise to the eukaryotic Cut/CDP family of proteins. PMID- 9332352 TI - Improved EBV-based shuttle vector system: dicistronic mRNA couples the synthesis of the Epstein-Barr nuclear antigen-1 protein to neomycin resistance. AB - Use of EBV-based vector systems has been limited by the requirement to generate EBNA+ cells which are 'permissive' for replication of an oriP-vector. In current constructs, selectable marker and EBNA-1 are not always co-expressed. This is a significant problem since the EBNA-1 gene product can be toxic in some cell types and may be selected against. In this paper, we describe a gene construct that overcomes this limitation. We have exploited the piconaviral internal ribosome entry site to allow the genes for Epstein-Barr nuclear antigen-1 and G-418 resistance to be transcribed as a dicistronic fusion mRNA under the control of the phosphoglucokinase promoter. This construct can be routinely integrated into human cell lines. The presence of EBNA-1 protein was reflected by a large increase in transfection frequencies (1000-fold) using an oriP-based vector which was shown to replicate stably in these cells with no apparent gross rearrangements detected after 8 weeks in culture. Using this system, G-418 resistance should directly reflect integration, as well as expression of the EBNA 1 gene, which, in turn, increases transfection frequencies and stability of EBV based vector systems and should result in its increased use. PMID- 9332353 TI - Genomic characterization of members of the Bet v 1 family: genes coding for allergens and pathogenesis-related proteins share intron positions. AB - Bet v 1, the major birch pollen allergen, is a member of a multigene family; a number of isoforms and homologous proteins from closely related species (alder, hazel and hornbeam) has been isolated and their cDNAs cloned and characterized. Genomic clones coding for Bet v 1 and homologues from apple and hazel were isolated and sequenced. Some of these clones contained intervening sequences. The exon-intron formation is highly conserved throughout this family of pathogenesis related proteins in dicot plants and is also found in Aopr1 (Asparagus officinalis), a monocol species. Phylogenetic analysis suggested a possible common origin of the intron position in these homologous proteins at codon 62 in various families of flowering plants, including Fagaceae, Rosaceae and Apiaceae. This conserved 'proto-splice site' may point to a structure/function relationship. A conserved sequence motif (P-loop) was also found in all members of this protein family. Moreover, there is a certain degree of sequence similarity among the proteins derived from various species throughout the dicots and the only monocot examined. This fact is reflected by cross-reactivity from monoclonal and polyclonal antibodies raised against Bet v 1. PMID- 9332355 TI - Sequence and transcript analysis of macronuclear histone H2B genes from Euplotes crassus. AB - Two 1.5-kb macronuclear chromosomes bearing histone H2B genes from the ciliated protozoan Euplotes crassus were cloned and sequenced. Although the noncoding sequences on these macronuclear chromosomes are very different, the genes encode an identical 113-aa histone H2B protein that has a shortened N-terminus and a highly conserved C-terminus relative to histone H2B proteins in other organisms. Primer extension was used to determine the transcription start points. Northern analysis shows that the abundance of H2B mRNA changes relative to DNA replication periods during the sexual phase of the life cycle. Analysis of 3' RACE products indicates that the H2B genes are coexpressed. PMID- 9332354 TI - Sequencing of the cholesteryl ester transfer protein 5' regulatory region using artificial transposons. AB - We have isolated and sequenced genomic clones encompassing more than 5 kb of the 5' flanking region of the cholesteryl ester transfer protein gene. This region contains multiple Alu repeats, a Mermaid repeat, and an extensive GA repeat, which made sequencing exceedingly difficult. To circumvent the problems that these repeats posed to traditional sequencing methodologies, we employed a novel transposon-facilitated technique, which greatly simplified sequencing of regions that had been difficult to accomplish otherwise. We utilized the artificial transposon, AT-2, a Bluescript derivative containing the dhfr gene and unique primer sites at both ends of the insertion DNA. Integration of the transposon occurred efficiently and covered the entire region of interest. Analysis of the sequence indicates a number of potential regulatory factor binding sites upstream of the previously characterized minimal promoter. The 5.7-kb regulatory region confers significant transcriptional activation in a conditionally transformed mouse hepatocyte line as compared to a minimal 137-bp promoter fragment. In addition, a tetranucleotide repeat of variable length that may provide a useful genetic marker has been identified 2 kb upstream of the CETP transcriptional start site. PMID- 9332356 TI - Cloning and analysis of cDNA encoding murine pinin. AB - Pinin is a cell junction-associated protein involved in the stabilization of the desmosome-intermediate filament complex in various epithelial tissues. Utilizing a cDNA probe derived from canine pinin, we isolated overlapping cDNA clones encoding murine full-length pinin. The total cDNA contained an open reading frame of 2175 nucleotides coding for 725 amino acids as well as a 3'- and a 5' untranslated regions of 620 and 18 nucleotides, respectively. The overall predicted amino acid sequence of mouse pinin displayed strong identities to those of canine and human pinin, with the exception of a stretch of 38 amino acids which were found to be deleted in mouse pinin. There were several discernible domains found within mouse pinin. These included three coiled-coil domains, a small stretch of glycine loops, a short glutamine-proline-rich domain and a polyserine domain. PMID- 9332357 TI - The cDNA cloning and transient expression of a chicken gene encoding a follicle stimulating hormone receptor. AB - A clone, pcFSHR, containing a 3.1-kb insert was isolated from a cDNA library of chicken ovarian follicles by screening with an RT-PCR-generated cDNA probe for a putative N-terminal half-region of chicken FSH-R. The deduced amino acid sequence of pcFSHR exhibits about 84% identity to those of mammalian FSH-R and about 70% to those of mammalian LH-CG-R. Northern blot analysis for total RNA preparations from several chicken tissues revealed that transcripts detected with pcFSHR were present exclusively in ovary and testis. In a chicken ovary, the transcripts were detected in granulosa but not in theca cells. A radioligand receptor assay for the human 293 cells transfected with an expression vector containing the insert of pcFSHR demonstrated the production of a receptor which showed about 10-fold higher affinity for chicken FSH than for chicken LH. The affinity for chicken FSH was significantly higher than for human FSH. In consistency with the ligand specificity of the receptor, a significantly higher level of intracellular accumulation of cAMP was detected when the transfected cells were treated with chicken FSH than with chicken LH. From these results, we conclude that pcFSHR is a cDNA clone for the chicken FSH-R. PMID- 9332358 TI - Chromosome assignment of aberrant NotI restriction DNA fragments in primary hepatocellular carcinoma. AB - DNA aberrations in human hepatocellular carcinoma (HCC) were studied by two dimensional DNA electrophoresis analysis. Five intensified and 60 dwindling spots were detected recurrently in the two-dimensional profile which showed about 3000 restriction DNA fragments as distinctive spots. We assigned these aberrant spots to chromosomes, using the chromosome-assigned two-dimensional profile. Four of the five intensified, and 53 of the 60 dwindling spots were given chromosome assignments. Intensified spots were assigned to chromosomes 5, 6, 9 through 12, 16 and 18. Among the dwindling spots, the highest incidence of aberrations was found on chromosome 16, followed by 9 through 12 and chromosome 2. No aberrations were detected in chromosomes 7, 21, 22 or Y. PMID- 9332359 TI - Functional and structural features of the holin HOL protein of the Lactobacillus plantarum phage phi gle: analysis in Escherichia coli system. AB - Lactobacillus plantarum phage phi gle has two consecutive cell lysis genes hol lys (Oki et al., 1996b). In the present study, functional and structural properties of the hol protein (Hol) were characterized in Escherichia coli. Electron microscopic examinations showed that hol under plac in E. coli XL1-Blue injured the inner membrane to yield empty ghost cells with the bulk of the cell wall undisturbed. Northern blot analysis indicated that hol-lys genes under plac were co-transcribed, although the amount of hol transcript was larger than that of lys, ceasing via an apparently rho-independent terminator just downstream of hol. However, deletion and/or fusion experiments suggested that: (1) the N terminal half of phi gle Hol composed of three putative transmembrane domains may be responsible for interaction with membrane; (2) the N-terminal end (five amino acids) seems nonessential; and (3) the C-terminal half containing charged amino acids appears to be involved in proper hol function. These results suggest that phi gle Hol is a member of the lambdoid holin family, but divergent in several properties from lambda holin. PMID- 9332360 TI - Structure of a fish (Oncorhynchus mykiss) vitellogenin gene and its evolutionary implication. AB - In this paper we describe the first complete structure of a fish vitellogenin gene. A 22 kb genomic region from rainbow trout (Oncorhynchus mykiss) was cloned and analysed. This region was shown to contain two tandemly arranged vitellogenin genes. Both genes are 98.7% similar, indicating that they result from a recent local duplication. The complete sequence encoding one of the two genes was determined and the gene organization was established. The gene is 10.3 kb long and has 34 exons, it lacks one exon compared to amphibian and avian vitellogenin genes. Exons 22 and 23 of the Xenopus and chicken genes were shown to be merged into a single exon in the trout genome. Other splicing sites appeared highly conserved between the three vertebrate genes. In contrast, little similarity between invertebrate and vertebrate vitellogenin genes was observed with respect to the number and organization of introns. The comparison of 17 independent invertebrate splicing sites with the 34 vertebrate sites indicated that a few sites are probably ancient. However, most of the splicing junctions compared appeared unrelated. Results suggest that vitellogenin genes have been reshaped through multiple insertions and deletions of intervening sequences during evolution. PMID- 9332361 TI - Cloning and characterization of a gene encoding an actin-related protein in Chlamydomonas. AB - The genomic sequence of an actin-related gene in Chlamydomonas reinhardtii has been determined. The deduced amino acid sequence of this gene shares a 63.9% identity with that of a recently reported conventional actin-encoding gene in C. reinhardtii. Phylogenetic analysis shows that the product of this actin-related gene does not fit into conventional actin or any major actin-related protein categories. The actin-related gene in C. reinhardtii contains seven introns in the coding region and, as described for the conventional actin gene, it contains several sequences similar to the 'tub box' sequence motif in its 5'-upstream region. Southern blot analysis of the gene shows a hybridization pattern different from that of the conventional actin gene, indicating that these genes are distinct from one another. Northern blot analysis of poly(A)+RNA shows the messages of the two genes to be very similar in size, yet the message level of the actin-related gene is significantly lower than that of the conventional actin gene. PMID- 9332362 TI - Isolation and sequence analysis of the cDNA encoding subunit C of human CCAAT binding transcription factor. AB - Sets of cDNA clones corresponding to genes with different expression specificity, 'brain-specific', 'common', 'liver-specific', were identified by differential hybridization of a human fetal brain cDNA library with total cDNA probes of human fetal brain and human fetal liver. Nucleotide sequence analysis revealed that one of the 'common' clones contained the cDNA encoding subunit C of human CCAAT binding transcription factor. The isolated human CBF-C cDNA is 1977 nt long and consists of the full-length 3'-untranslated region (781 nt) with a poly (A) tail at the 3' end 185 nucleotides of 5'-untranslated region and the open reading frame (1011 nt), encoding a 337-aa protein with 91.7% homology with the translated region of rat CBF-C cDNA. PMID- 9332363 TI - Cloning and sequencing of the dnaK and grpE genes of Legionella pneumophila. AB - A 4.4-kb DNA fragment from Legionella pneumophila (Lp) was isolated, which could complement an Escherichia coli (Ec) dnaK ts mutant, HC4102. Nucleotide sequence analysis of the region revealed two complete open reading frames (ORFs) encoding both a predicted DnaK protein of 644 aa and a predicted GrpE protein of 199 aa, and also the 5'-end of the predicted dnaJ gene organized in the order of grpE dnaK-dnaJ. Consensus heat shock (HS) promoter sequences were identified upstream of the start of both grpE and dnaK transcripts. However, no obvious promoter sequences were detected upstream of dnaJ. The transcription start points of grpE and dnaK were determined by primer extension analysis and the amount of each of the transcripts increased four- to eightfold after HS. PMID- 9332364 TI - Disrupted cholecystokinin type-A receptor (CCKAR) gene in OLETF rats. AB - OLETF rats develop hyperglycemia, hyperinsulinemia and mild obesity, which is characteristic of human non-insulin-dependent diabetes mellitus (NIDDM). We cloned and sequenced the cholecystokinin type-A receptor (CCKAR) gene in the rats. Comparing the DNA sequences of the OLETF CCKAR gene and LETO CCKAR gene, normal gene, we found a deletion in the OLETF gene, 6847 bases in length, which was flanked by two 3-base-pair direct repeats (5'-TGT-3') at positions -2407/ 2405 and 4441/4443, numbered according to the LETO gene sequence, one of which was lost. The promoter region, the first and second exons were missing in the mutant. The region upstream and downstream of the deletion, including exons 3, 4 and 5, was conserved between the two strains, and did not contain any base changes. We found that the gene mapped to chromosome 14 in rats. OLETF rats are the naturally occurring knockout animals with the homozygously disrupted CCKAR gene. PMID- 9332365 TI - The human PD-1 gene: complete cDNA, genomic organization, and developmentally regulated expression in B cell progenitors. AB - We report the complete cDNA sequence and the genomic structure of the human PD-1 homologue. An analysis of the expression pattern of the human PD-1 gene (hPD-1) and the murine PD-1 gene (mPD-1) in developing bone marrow B-lineage cells was also undertaken. The full length hPD-1 cDNA is 2106 nucleotides long and encodes a predicted protein of 288 amino acid residues. The hPD-1 and mPD-1 genes share 70% homology at the nucleotide level and 60% homology at the amino acid level. Four potential sites for N-linked glycosylation are conserved, as are a stretch of amino acids between two cysteine residues resembling a V-set immunoglobulin domain, and another region containing a motif similar to an immunoreceptor tyrosine-based inhibitory motif. Isolation of the genomic locus of the hPD-1 gene reveals that the gene is composed of five exons located on human chromosome 2 at band q37. The 5' flanking region lacks TATA and CAAT cis-acting elements, but includes a number of potential transcription factor binding sites and a dominant transcription start site. The mPD-1 gene was preferentially expressed in pro-B cells from murine adult bone marrow. Although hPD-1 was not preferentially expressed in pro-B cells from human fetal bone marrow, treatment of isolated pro B cells with interleukin-7 resulted in a dramatic increase in expression. These data suggest that PD-1 may play a role in B-cell differentiation during the pro-B cell stage. PMID- 9332366 TI - The human glutaredoxin gene: determination of its organization, transcription start point, and promoter analysis. AB - A genomic clone for the human glutaredoxin gene was isolated and sequenced. An intron was located within the coding region and began 211 nt downstream of the initiator codon. Except for this intron, the genomic sequence shares 100% identity to the published glutaredoxin cDNA sequence. A second intron was located in the 3' UTR 6 bp downstream of the terminator codon. The tsp of the glutaredoxin gene was determined by primer extension and confirmed by S1 mapping analysis. Analysis of the 5'-flanking region of the gene revealed that the promoter sequences TATA and CCAAT were 30 and 160 bp upstream, respectively, from the tsp. Other potential transcription factor binding sites included NF-E1, HNF 5, P2II and AP-1. Glutaredoxin promoter constructs inserted into a reporter plasmid for firefly luciferase were transfected into fibroblasts, and luciferase activity was 8-10-fold higher compared with controls lacking glutaredoxin promoter. These data indicate that the promoter region of the isolated glutaredoxin gene is functional. PMID- 9332367 TI - Cloning of a human multispanning membrane protein cDNA: evidence for a new protein family. AB - We report the cloning of a human cDNA encoding a protein of calculated 68.8 kDa molecular mass, named hMP70. The deduced protein sequence shows a large N terminal hydrophilic part and a C-terminal part with nine putative hydrophobic regions characteristic of integral transmembrane domains. Computer searches with sequence databases revealed homologies with three complete yeast proteins and with at least 19 human, 10 plant and one nematode short unidentified protein sequences translated from Expressed Sequence Tags (ESTs). Remarkably, this hMP70 protein retains between 27 and 31% overall sequence identity with the yeast proteins. We propose that hMP70 and related genes have evolved from a common ancestral gene and form a new multispanning membrane protein family which we call the MP70 protein family. Gene expression of hMP70 appears to be ubiquitous, as the mRNA is detectable in all human tissues analysed so far, as shown by Northern blot analysis. Furthermore, a protein of about 70 kDa is detectable in different mammalian cell lines, as shown by immunoblot analysis. From its widespread expression and conservation from yeast, plants to mammals, it is likely that hMP70 has a fundamental biological function in the cell. PMID- 9332368 TI - Nucleotide sequences and gene organization of TaqI endonuclease isoschizomers from Thermus sp. SM32 and Thermus filiformis Tok6A1. AB - Eight TaqI isoschizomer genes, two from Yellowstone National Park, one from Japan, two from New Zealand, two from Portugal, and one from the Azores (1000 miles west of Portugal), were PCR-amplified and sequenced. Sequence alignment of isoschizomers isolated from close geographical locations shows identical or almost identical protein sequences, while isoschizomers from distant sites demonstrate considerable diversity, ranging from 54 to 75% in amino acid identity. Accordingly, these isoschizomers were arranged into four geographical groups, i.e., USA as represented by Thermus aquaticus YT1, Japan by Thermus thermophilus HB8, New Zealand by Thermus filiformis Tok6A1, Portugal by Thermus sp. SM32. The complete ORFs of two new representative genes, tfiTok6A1I and tsp32IR, were obtained by bubble PCR. Unlike M . TaqI-R.TaqI and M . TthHB8I-R . TthHB8I which exhibit an unusual 13-codon overlap, the methylase and endonuclease genes are each separated by 15 nucleotides in the TfiTok6A1I and Tsp32IR restriction-modification systems. Phylogenetic analysis suggests that initially TfiTok6A1I diverged from a common ancestor, then Tsp32IR branched out, and finally TaqI and TthHB8I diverged from each other during evolution. PMID- 9332369 TI - Expression, genomic structure and high resolution mapping to 19p13.2 of the human smooth muscle cell calponin gene. AB - Smooth muscle cells (SMC) express a battery of cell-restricted differentiation genes, many of which are down-regulated during the course of vascular disease. Here, we present the mRNA expression, genomic structure and chromosomal mapping of the gene encoding human smooth muscle cell calponin (SMCC). Human SMCC transcripts are restricted to tissues and cells of SMC origin and, in the latter case, appear to be uniquely controlled in two distinct human SMC lines of uterine and aortic origin. Restriction mapping. Southern blot and PCR analysis of a 70-kb human bacterial artificial chromosome (BAC) revealed a genomic structure (seven exons spanning > 11 kb) very similar to that reported for the mouse SMCC gene. Using a variety of human-rodent somatic cell hybrid and radiation hybrid mapping panels, the human SMCC gene was mapped to a genomic interval of less than 1.32 Mb in 19p13.2. These results provide new information concerning the regulation of SMCC gene expression and demonstrate the utility of two human SMC lines for the further characterization of this gene's expression control. The identification of a BAC harboring the entire human SMCC locus represents an important reagent for future analysis of SMCC regulatory sequences. Finally, the localization of SMCC to a defined genomic interval will facilitate an analysis of its potential as a candidate gene for disease phenotypes mapping to 19p13.2. PMID- 9332370 TI - A CHD1 gene is Z chromosome linked in the chicken Gallus domesticus. AB - Chromo-helicase-DNA binding 1 (CHD1) is a conserved protein with a putative role in chromatin architecture. Single homologues have been found in mouse, Drosophila and yeast. In birds the situation is different as they possess two homologues. One is known to be W-linked, we show the second, closely related gene is linked to the Z sex chromosome. The basic structure of the Z-linked gene is similar to the homologous genes, however, it does possess an additional, internal 88 amino acid hydrophilic domain, rich in glutamic acid and lysine. Studies on pairs of genes sex-linked in mammals suggests rapid divergence of DNA sequence and function. We suggest the DNA sequences of CHD-W and CHD-Z do not follow this pattern. PMID- 9332371 TI - Molecular cloning and characterization of a cDNA encoding a bovine butanediol dehydrogenase. AB - Using a polyclonal antibody against a bovine brain 30-kDa protein (p30), we isolated from a lambda gt11 bovine brain expression library a cDNA that codifies a protein with an apparent molecular mass of 30 kDa. The cDNA nucleotide sequence contained a unique open reading frame encoding a 26.7 kDa polypeptide. The 257 amino acids deduced sequence showed a significant homology with several dehydrogenases, mainly with a bacterial acetoin reductase (62%). The cloned cDNA identity was confirmed by the determination of acetoin reductase activity in lysogens of lambda phage constructions containing the full length cDNA. The results described in this report are to our knowledge the first molecular characterization of a 2,3-butanediol dehydrogenase in mammals. PMID- 9332372 TI - Characterization of two members of the Arabidopsis thaliana gene family, At beta fruct3 and At beta fruct4, coding for vacuolar invertases. AB - We have isolated and characterized two Arabidopsis thaliana cDNAs and their cognate genes, At beta fruct3 and At beta fruct4, encoding vacuolar forms of invertase. Our sequencing results showed that the gene At beta fruct3 is located downstream of the 3-ketoacyl-acyl carrier protein synthase III gene (AtKasIII). At beta fruct3 and 4 are functional and organized into seven exons and six introns with an identical organization. The At beta fruct3 and At beta fruct4 genes encode, respectively, polypeptides of 648 and 664 residues that contain all the characteristic hallmarks of vacuolar invertases. A. thaliana is the first plant of which both cell-wall (At beta fruct1 and At beta fruct2) and vacuolar (At beta fruct3 and At beta fruct4) genes are characterized. The same number of exons and introns is seen in the genes At beta fruct1, At beta fruct3 and At beta fruct4 as well as in all other invertase genes described to date. However, the position of the third intron is different in At beta fruct3 and At beta fruct4. At beta fruct2 shows a different organization. A neighbour-joining distance tree shows that the A. thaliana vacuolar invertases described here are, as expected, more closely related to vacuolar invertases from other plant species (e.g., carrot) than to the A. thaliana cell-wall invertases. The evolution of plant invertase genes from a common ancestral gene is discussed. Our results demonstrate that in A. thaliana, at least two genes encoding vacuolar invertases are expressed during the development of the plant. Southern blot hybridization experiments suggest the presence of one copy of, respectively, At beta fruct3 and At beta fruct4 per haploid genome, and Northern blot analysis demonstrates that vacuolar invertase genes are highly expressed in stems, roots, flowers and at very low levels in mature leaves. PMID- 9332373 TI - Oligodeoxyribonucleotide length and sequence effects on intramolecular and intermolecular G-quartet formation. AB - The potential of guanine-rich oligodeoxyribonucleotides (oligos) as nucleic acid drugs is increasingly being investigated, for example, as aptamers against heparin-binding proteins and as purine-motif triplex-forming oligos. However, G rich oligos can be very polymorphic under physiological conditions, often with the resulting structures possessing vastly different functional capabilities. To better understand the intrinsic oligo parameters that affect their structure, we used nondenaturing gel electrophoresis to investigate a series of G-rich oligos derived from the sequence 5'-TGGGTGGGGTGGGGTGGGT for their abilities to self associate through G-quartet formation. From these studies the following observations could be made: (1) oligos containing four clusters of three or more contiguous Gs readily associated intramolecularly but did not associate intermolecularly; (2) intermolecular dimerization was the preferred mode of interaction when one of the oligos contained only two G clusters; and (3) T-rich extensions promoted multimerization of oligos into still higher-order species. PMID- 9332374 TI - Molecular cloning of chicken FTZ-F1-related orphan receptors. AB - FTZ-F1 is a member of the orphan nuclear receptors, which belongs to the steroid hormone receptor superfamily, and plays a role in the blastoderm and nervous system development in Drosophila. Recently, several FTZ-F1 family genes have been cloned in several species. SF-1/Ad4BPs have been identified as master regulators controlling steroidogenic P-450 genes in mammals and are considered to be the mammalian homologues of FTZ-F1. Moreover, SF-1/Ad4BP plays a critical role in the sexual differentiation of gonads in mammals. In vertebrates, except for mammals, the functional homologue of SF-1/Ad4BP has not been identified before. Herein, we cloned two chicken cDNAs (OR2.0 and OR2.1), which encode putative FTZ-F1 family receptors, by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). OR2.1 consists of 3255 bp, is expressed in the adrenal glands and gonads, and is considered to be the chicken counterpart of mammalian SF-1/Ad4BP. However, OR2.0 consists of 2945 bp, is expressed in the livers and the adrenal glands, and is considered to be the chicken counterpart of mouse LRH-1, which is a member of the FTZ-F1 family in mammals. PMID- 9332375 TI - Specific isolation of human rDNA genes by TAR cloning. AB - Selective cloning of human DNA in YACs from monochromosomal human/rodent hybrid cells lines and radiation hybrids can be accomplished by transformation associated recombination (TAR) between Alu-containing vector(s) and human DNA in yeast. We have expanded this approach to the specific isolation of repetitive genes from the human genome. Highly selective isolation of human rDNA was accomplished using total human DNA and a pair of differentially marked linear TAR cloning vectors where one contained a small fragment of a human rDNA repeat and the other had an Alu repeat as targeting sequences. About half the transformants that acquired both vectors markers had YACs with human rDNA inserts. These results suggest that TAR can be applied to the general isolation of gene families and amplified region from genomic DNAs. PMID- 9332376 TI - Molecular cloning and expression of a porcine chondrocyte nucleotide pyrophosphohydrolase. AB - The porcine 127-kDa nucleotide pyrophosphohydrolase (NTPPHase) had been previously purified from the conditioned culture media of porcine articular cartilage. Protein sequencing of an internal 61-kDa proteolytic fragment of NTPPHase (61-kDa NTPPHase) determined the 26 N-terminal amino acids. This sequence was used to amplify a DNA fragment, which was used as a probe to clone the gene encoding the 61-kDa NTPPHase from a porcine chondrocyte cDNA library. DNA sequence analysis showed the cDNA insert to be 2509 bp, corresponding to a predicted open reading frame (ORF) encoding 599 amino acids. The 26 N-terminal amino acids of the 61-kDa NTPPHase were located within the ORF immediately downstream of a putative protease recognition region, RRKRR. This is consistent with this cDNA insert representing an internal proteolytic fragment of the full length 127-kDa NTPPHase. BLAST and FASTA analysis confirmed that the deduced amino acid sequence of 61-kDa NTPPHase was unique and did not possess a high degree of homology to sequence in the non-redundant protein and nucleotide databases. Proteins that possess limited homology (< 17%) with the 61-kDa NTTPPHase include several prokaryotic and eukaryotic ATP pyrophosphate-lyases (adenylate cyclase). Northern blot analysis of porcine chondrocyte RNA showed that the DNA encoding the 61-kDa NTPPHase hybridized to a single 4.0-kb RNA transcript. This DNA probe also hybridized to a single species of human chondrocyte RNA. Expression of a 61-kDa protein was detected by coupled in-vitro transcription/translation. Western blot analysis of this in-vitro transcription/translation reaction detected a 61-kDa protein, using an antibody raised against the peptide sequence that was originally used to clone the 61-kDa NTPPHase. These data indicate the successful in-vitro cloning and expression of the porcine chondrocyte 61-kDa NTPPHase. Future studies that utilize the gene encoding the 61-kDa NTPPHase may allow the characterization of the role of NTPPHase in calcium pyrophosphate dihydrate (CPPD) crystal deposition disease. PMID- 9332377 TI - Molecular cloning and expression of a rat cDNA encoding 5-aminoimidazole-4 carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase. AB - The cDNA of a 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (AICARFT/IMPCHase) was isolated from rat liver RNA by reverse transcription and the polymerase chain reaction (PCR). The rat AICARFT/IMPCHase cDNA included 1928 bp containing a coding region of 1779 bp for a 592-amino acid polypeptide (Mr = 64 200). Rat and human AICARFT/IMPCHase cDNAs show 84 and 91% homology at the nucleotide and amino acid sequence level, respectively. The protein produced by the rat cDNA using pET-expression system catalysed the penultimate and final steps of de novo purine biosynthesis. Northern analysis identified a 2.8-kb AICARFT/IMPCHase mRNA and the level of the AICARFT/IMPCHase transcripts increased markedly at 24 h after partial (70%) hepatectomy. PMID- 9332378 TI - Isolation of the cDNA and characterization of mRNA expression of ribosomal protein S19 from the soft-shell clam, Mya arenaria. AB - Ribosomal proteins contribute to the regulation and activity of ribosomes, and hence, the translational activity of the cell. Aberrant expression of ribosomal proteins has been linked to certain pathological conditions such as neoplasms. We have isolated and characterized a cDNA for the ribosomal protein (rp) S19 from a marine bivalve, the soft-shell clam (Mya arenaria), and we have examined its pattern of mRNA expression in the ovary and testis. The S19 cDNA contains a 450 nucleotide (nt) open reading frame (ORF), flanked by 89 nt and 26 nt of 5' and 3' untranslated regions, respectively. Probes synthesized from the S19 cDNA recognize a single transcript of approximately 550 nt in four different tissues. The predicted amino acid sequence from the ORF exhibits 58% identity with human and rat S19. Southern analysis of genomic DNA suggests that M. arenaria may have multiple copies of S19, a feature that is more similar to vertebrate than invertebrate rp genes. Expression of S19 mRNA in both ovary and testis was elevated throughout gametogenesis until after spawning, when a decrease in S19 message was observed. A comparison of S19 mRNA levels in post-spawn animals revealed a trend of elevated expression in ovaries and testes affected by a gonadal neoplasm, indicating that S19 may be a useful molecular marker for the pathological condition. PMID- 9332379 TI - Chicken repeat 1 (CR1) elements, which define an ancient family of vertebrate non LTR retrotransposons, contain two closely spaced open reading frames. AB - Chicken repeat 1 (CR1) elements comprise a family of non-long terminal repeat (LTR) retrotransposons that have several noteworthy features. For example, whereas most other non-LTR elements have poly(A) tracts or other simple A-rich repeats at their 3' ends, the 3' ends of CR1 elements conform to the consensus [(CATTCTRT)(GATTCTRT)1-3]. CR1 elements also display an unusual bias for severe 5' truncations: only approx. 30 (out of a total of approx. 30 000) CR1 elements in the chicken genome include significant portions of the pol-like open reading frame (ORF) that we previously identified and partially sequenced [Burch et al. (1993) Proc. Natl. Acad. Sci. USA 90, 8199-8203]. In the present study we derived a consensus sequence for this entire ORF (ORF2) as well as an upstream ORF (ORF1) and part of a 5' untranslated region (UTR). The conceptual translation product of ORF2 is predicted to contain an endonuclease domain in addition to a reverse transcriptase domain. These results suggest that CR1 elements retrotranspose using a "nick and prime" mechanism similar (but not identical) to other families of non-LTR elements. PMID- 9332380 TI - The myeloperoxidase gene proximal enhancer directs hematopoietic-specific expression in transgenic mice. AB - The myeloperoxidase (MPO) gene is expressed specifically in immature myeloid cells. The MPO gene includes a promoter proximal enhancer which is coincident with DNaseI hypersensitive chromatin sites and is specifically active in myeloid cell lines. We developed transgenic murine lines in which 1.3 kb of murine MPO proximal 5' flanking region DNA was linked to a TATAA homology and RNA initiation site derived from the HSV-TK promoter and to a luciferase reporter (MPOTKLUC). In each of six founder lines, high-level luciferase activity was evident in marrow, thymus and spleen. Modest- to high-level luciferase expression was also evident in brain and in the heart in several of the lines, and luciferase activity was at or near background levels in lung, liver, kidney, stomach, colon, bladder, skeletal muscle, skin and small intestine in all of the MPOTKLUC transgenic mice. Within marrow cells, luciferase activity was evident in myeloid (GR-1+), B lymphoid (B220+) and T-lymphoid (CD4+) cells. Additional regulatory regions, thus, may be required to further restrict MPO gene expression to immature myeloid cells. PMID- 9332381 TI - A Drosophila muscle-specific gene related to the mouse quaking locus. AB - We have characterized a novel muscle-specific gene of Drosophila melanogaster, defined by enhancer trap strain 24B of Brand and Perrimon (1993). We show that transcripts of the gene accumulate within presumptive mesoderm and persist within developing muscles, strongly suggesting that the encoded protein is involved in muscle cell determination and differentiation. cDNA sequences reveal that the Drosophila protein is similar to quaking (64% identity over 210 amino acids), a protein essential for mouse embryogenesis, and gld-1 (53% identity over 162 amino acids) a germ-line-specific tumor suppressing protein of the nematode, Caenorhabditis elegans. We demonstrate that the Drosophila gene resides within the 93F chromosome subdivision, and describe its physical map. Finally, we have used the gene, which we have named quaking-related 93F (qkr93F), to identify a family of closely related KH domains. PMID- 9332382 TI - The human mitochondrial elongation factor tu (EF-Tu) gene: cDNA sequence, genomic localization, genomic structure, and identification of a pseudogene. AB - The human mitochondrial elongation factor Tu (EF-Tu) is nuclear-encoded and functions in the translational apparatus of mitochondria. The complete human EF Tu cDNA sequence of 1677 base pairs (bp) with a 101 bp 5'-untranslated region, a 1368 bp coding region, and a 207 bp 3'-untranslated region, has been determined and updated. The predicted protein from this cDNA sequence is approximately 49.8 kDa in size and is composed of 455 amino acids (aa) with a putative N-terminal mitochondrial leader sequence of approximately 50 aa residues. The predicted amino acid sequence shows high similarity to other EF-Tu protein sequences from ox, yeast, and bacteria, and also shows limited similarity to human cystolic elongation factor 1 alpha. The complete size of this cDNA (1677 bp) obtained by cloning and sequencing was confirmed by Northern blot analysis, which showed a single transcript (mRNA) of approximately 1.7 kb in human liver. The genomic structure of this EF-Tu gene has been determined for the first time. This gene contains nine introns with a predicted size of approximately 3.6 kilobases (kb) and has been mapped to chromosome 16p11.2. In addition, an intronless pseudogene of approximately 1.7 kb with 92.6% nucleotide sequence similarity to the EF-Tu gene has also been identified and mapped to chromosome 17q11.2. PMID- 9332383 TI - An epitope tagged mammalian/prokaryotic expression vector with positive selection of cloned inserts. AB - A dual eukaryotic/prokaryotic expression vector has been developed which combines the features of positive selection for cloned inserts along with the production of an epitope-tagged cDNA insert by transient transfection in mammalian cells as well as high level induced expression in E. coli cells harbouring T7 RNA polymerase. This vector, pZilch, has two MCSs flanking a mutant E. coli phenylalanyl-tRNA synthetase gene, pheS, which when expressed in combination with the phenylalanine analog p-CI-Phe, results in termination of host cell protein synthesis. Cloning of inserts using unique sites in the flanking MCS regions results in loss of the pZilch pheS allele and hence permits growth of colonies harbouring recombinants on p-Cl-Phe plates. Additional features of the vector include an optimal Kozak consensus sequence for high level eukaryotic cell expression and an efficient prokaryotic translation initiation site in frame and downstream from the eukaryotic initiation site. Recombinant proteins can be produced with an N-terminal FLAG epitope which can be removed via a specific protease cleavage site. Flanking T7 and SP6 RNA polymerase promoter sites permit in vitro transcription and translation of cloned inserts. A derivative of the vector has also been constructed enabling nuclear accumulation of the tagged proteins via an SV40 nuclear localisation signal upstream of the 5' MCS. PMID- 9332384 TI - An alfalfa rubisco small subunit homologue shares cis-acting elements with the regulatory sequences of the RbcS-3A gene from pea. AB - A genomic clone of RbcS was isolated from an alfalfa (Medicago sativa L. cv. Apica) genomic library and characterized. Although this clone has structural features similar to a functional gene, the second exon is interrupted by a stop codon and thus is not fully translatable in the plant. Sequence analysis of the 5' and 3' noncoding regions of RbcSK-1A showed a high sequence homology to the flanking sequences of the RbcS-3A gene from pea. The regions of homology contain many important cis-regulatory elements shown to be essential for regulation of the RbcS-3A gene in pea. The promoter of this alfalfa rubisco clone was used in a translational fusion to test its ability to control the expression of the GUS reporter gene in an homologous nuclear background. High levels of GUS enzyme activity were recorded. These strong levels are comparable to some exceptionally high levels produced in other studies following the use of photosynthesis gene promoters in fusions with the GUS reporter gene. PMID- 9332385 TI - Cloning and characterization of the gene encoding PspPI methyltransferase from the Antarctic psychrotroph Psychrobacter sp. strain TA137. Predicted interactions with DNA and organization of the variable region. AB - The gene (pspPIM) encoding the PspPI DNA methyltransferase (MTase) associated with the PspPI restriction-modification (R-M) system (5'-GGNCC-3') of Psychrobacter species TA137 has been cloned and expressed in E. coli, and its nucleotide (nt) sequence has been determined. The coding region was 1248 nt in length and capable of specifying a 46826-Da protein of 416 amino acids (aa). The predicted sequence of the MTase protein displays ten sequence motifs characteristic of all prokaryotic m5C-MTases and shows the highest similarity to other MTases that methylate the GGNCC sequence, namely M . Eco47II and M . Sau96I. All three MTases methylate the internal cytosine within their recognition sequence. Sequence similarities between M . PspPI and its isospecific M . Eco47II and M . Sau96I as well as with four other m5C-MTases that methylate the related GGWCC sequence, namely M . SinI, M . HgiCII, M . HgiBI, M . HgiEI have been also found within the variable region of these proteins. On the basis of structural information from M . HhaI and M . HaeIII, several M . PspPI residues that are expected to interact with DNA can be predicted. Furthermore, an organization of the variable region of m5C-MTases into two segments exhibiting a pattern of conserved residues and a considerable degree of structural homologies is described. PMID- 9332386 TI - Cloning and sequence analysis of the immediate promoter region and cDNA of porcine granulocyte colony-stimulating factor. AB - Granulocyte colony-stimulating factor (G-CSF) is a cytokine that stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophil/granulocyte lineage. Recombinant G-CSF (rG-CSF) is routinely used in the prevention of chemotherapy-induced neutropenia and in the setting of bone marrow transplantation. Chronic idiopathic and congenital neutropenic disorders also show improvement after rG-CSF injections. Applications of either rG-CSF or G CSF gene transfected cells into mice give rise to leukocytosis, which can be measured easily. This makes G-CSF a versatile tool for studying systemic effects of therapeutic proteins delivered by genetically modified cells in vivo. Although the biological activity of G-CSF is not species-specific, studies on long-term expression would require the use of species-identical proteins in order to avoid host immune reactions against the foreign gene product. Because of the physiological and immunological similarity of pigs and human, the pig has become an important large-animal model for biomedical research. We have therefore cloned porcine G-CSF cDNA from RNA isolated from pig PBLs. Pig G-CSF is a 195-amino-acid polypeptide that shares a high degree of homology to human (78%), murine (71%) as well as rat (68%) G-CSF. In contrast to human and murine, but not to rat G-CSF, a different ATG translation start codon is used, resulting in a shorter, but still functional signal sequence. PMID- 9332387 TI - Use of ordered deletions in genome sequencing. AB - Previous attempts to use the non-random approach for sequencing long DNA fragments have met with little success. As a result, nearly all genomic sequencing is done by the random (shotgun) approach, and the economy promised by the non-random approach has so far not materialized. Here we describe a simple system based on the use of ordered deletions that can be incorporated in the common strategies for genome sequencing. Long genomic fragments are cloned in the pAL-F cosmid and fragmented by digestion with specific restriction endonucleases. The digests are religated to subclone individual restriction fragments. The subclones are then subdivided by overlapping deletions and used for sequencing. We present the nucleotide sequences of two cosmid inserts from chromosome IV of Drosophila (containing the ci gene and the 5' end of the zfh-2 gene) that were determined by this method. This is the first report of successful sequencing of long genomic fragments by the use of overlapping deletions. Our calculations show that, with the present approach, sequence data can be acquired at a rate comparable to the shotgun approach but with significantly reduced numbers (approximately 30%) of sequencing runs. Hence, the use of ordered deletions should allow significant savings in both the amount and cost of sequencing work. PMID- 9332388 TI - Distinct cis-acting sequences are required for the germination and sugar responses of the cucumber isocitrate lyase gene. AB - Recent data have shown that distinct DNA sequence elements direct the germination and sugar responses of the cucumber (Cucumis sativus, L.) malate synthase (Ms) gene (Sarah et al. (1996) Mol. Gen. Genet., 250, 153-161). Such information is, however, lacking for the isocitrate lyase (Icl) gene which is coordinately regulated with Ms. Deletions from the 5' end of the Icl promoter were therefore created specifically to address this question. Analysis of expression in seeds of transgenic Nicotiana plumbaginifolia plants showed that whereas a promoter sequence of 2.9 kilobases (kb) produced a normal germination response, deletion to -1568 base pairs (bp) dramatically reduced this response. Examination of the sugar response employed a transgenic cucumber root system. In this case, the 2900 bp and 1568 bp promoters both gave a strong sugar response, but further deletion to -1367 bp eliminated the response. Therefore, the germination and sugar responses of the Icl gene require distinct cis-acting elements, located respectively upstream and downstream of -1568 bp. This observation is consistent with distinct signal transduction systems regulating gene expression in each case. PMID- 9332389 TI - Expression and sequence analysis of the Drosophila blastoderm-specific gene bsg25A. AB - By differential screening of a genomic library, we have cloned a gene expressed specifically during the blastoderm stage of Drosophila embryogenesis. Northern blot analysis and in situ hybridization to embryos reveal that the transcript is maximally expressed during the late syncytial blastoderm stage, disappears rapidly during the cellular blastoderm stage and is not detected at any other point in the Drosophila life cycle. On the basis of its temporally restricted expression and its polytene chromosomal map position at 25A1,2, we have designated this gene blastoderm-specific gene 25A (bsg25A). bsg25A encodes a novel protein of 23 kDa. PMID- 9332390 TI - Cloning of cDNA and expression of the gene encoding rat presenilin-2. AB - We have cloned the rat homologue of the presenilin-2 (PS-2) cDNA. PS-2 is responsible for chromosome 1-linked familial Alzheimer's disease. Sequence analysis predicted that the rat PS-2 encodes a 448 amino acid (aa) protein, and there was a very high degree of amino acid identity between rat and human PS-2 (95%). All the mutated codons in PS-2 and PS-1 in chromosome 1- or 14-linked familial Alzheimer's disease patients were conserved in rat PS-2. The expression of PS-2 was weaker than that of PS-1. The alternatively spliced short form of PS 2 mRNA, which was detected in human tissues was not detected in various rat tissues. During brain development, the expression level of both PS-2 and PS-1 increased but decreased in the adult. No remarkable change was observed in neural differentiation of PC12 cells. PMID- 9332391 TI - Isolation and nucleotide sequence of human liver glycogen debranching enzyme mRNA: identification of multiple tissue-specific isoforms. AB - Glycogen storage disease type III (GSD-III) is caused by a deficiency of glycogen debranching enzyme (AGL) activity. Patients are found to have deficient AGL activity in both muscle and liver, and also enzyme deficiency in the liver, but not in muscle. To determine the molecular basis of enzymatic variability in GSD III and to elucidate the mechanism for control of tissue-specific expression of AGL, we previously cloned and sequenced the human muscle AGL cDNA. Here we report the isolation and nucleotide sequence of liver AGL cDNA and the tissue distribution of the isoform mRNAs. The predominant form of human liver AGL cDNA (isoform 1) contained 400 bp of 5' untranslated region, 4596 bp of coding region, and 2371 bp of 3' untranslated region. The liver AGL mRNA sequence was identical to the previously published muscle sequence (isoform 5) for most of the length, except for the 5' end, in which the liver sequence diverged completely from the muscle sequence. The divergence began with the transcription start point and extended 82 nucleotides downstream from the translation initiation codon. Six isoforms of AGL mRNA were identified and sequenced from liver and muscle. These isoforms differed only at the 5' end. Tissue distribution studies showed that liver, kidney and lymphoblastoid cells expressed predominantly isoform 1; whereas muscle and heart expressed not only isoform 1, but also muscle-specific isoform mRNAs (isoforms 2, 3 and 4). Defining tissue-specific AGL isoform mRNAs is an important step toward understanding the molecular basis of enzymatic variability in GSD-III. PMID- 9332392 TI - Cloning of mouse FGF10 and up-regulation of its gene expression during wound healing. AB - We cloned the mouse homolog of FGF10, which was recently reported as a new member of the FGF family. The predicted molecular mass of this molecule is 23.6 kDa, and both nucleotide and amino acid sequences show high degrees of similarity with those of the rat. Examination of mouse FGF10 mRNA expression in various tissues and developmental stages by Northern hybridization revealed tissue- and developmental stage-specific expression of the gene. Similarly to the rat counterpart, mouse FGF10 mRNA (4.5 kb) was expressed relatively abundantly in embryos and the lung, and at much lower levels in brain and heart. In addition, a shorter transcript (1.3 kb) is expressed only in testis. Considering the high similarity in primary structure between FGF10 and FGF7 (known as keratinocyte growth factor; KGF), we also examined the gene expression of FGF10 during wound healing using a mouse model. FGF10 mRNA was highly induced 1 day after injury and decreased rapidly by 3 days. This suggests that FGF10 is a primary factor in the process of wound healing similarly to other growth factors such as TGF alpha and FGF7. PMID- 9332393 TI - First characterization of the phosphonoacetaldehyde hydrolase gene of Pseudomonas aeruginosa. AB - The phnX gene encoding the phosphonoacetaldehyde hydrolase (phosphonatase) from the Gram-negative bacterium Pseudomonas aeruginosa A237 has been cloned and its sequence determined. The open reading frame consists of 825 nucleotides specifying a protein of 275 amino acid residues corresponding to a predicted molecular weight of 29929. The deduced amino acid sequence of PhnX did not share significant amino acid sequence similarity with any other polypeptide. Expression of the phosphonoacetaldehyde hydrolase coding sequence in Escherichia coli under control of the E. coli tac promoter resulted in the production of enzymatically active protein with an affinity constant similar to that of the phosphonoacetaldehyde hydrolase purified from P. aeruginosa A237. This is the first nucleic sequence report of the phosphonoacetaldehyde hydrolase, an enzyme involved in the carbon-phosphorus bond cleavage. PMID- 9332394 TI - A reconstruction of the metabolism of Methanococcus jannaschii from sequence data. AB - The interpretation of the Methanococcus jannaschii genome will inevitably require many years of effort. This initial attempt to connect the sequence data to aspects of known biochemistry and to provide an overview of what is already apparent from the sequence data will be refined. Numerous issues remain that can be resolved only by direct biochemical analysis. Let us draw the reader's attention to just a few that might be considered central: (1) We are still missing key enzymes from the glycolytic pathway, and the conjecture is that this is due to ADP-dependency. The existence of glycolytic activity in the cell-free extract should be tested. (2) The issue of whether the Calvin cycle is present needs to be examined. (3) We need to determine whether the 2-oxoglutarate synthase (ferredoxin-dependent) (EC 1.2.7.3) activity is present. (4) The issue of whether cyclic 2,3-bisphosphate is detectable in the cell-free extracts needs to be checked. If it is, this result would confirm our assertion of the two pathways controlling synthesis and degradation of cyclic 2,3-bisphosphate. PMID- 9332448 TI - Protein kinase C and T cell function. PMID- 9332449 TI - IGF-1 stimulates synthesis of undersulfated proteoglycans and of hyaluronic acid by peritubular cells from immature rat testis. AB - The exposure of confluent peritubular (PT) cells from immature rat testis to insulin-like growth factor-1 (IGF-1) induced a time and dose-dependent increase of [35S]-sulfate and [3H]-D-glucosamine incorporations in newly synthesized proteoglycans (PG). This increased content of PG was the result of an enhancement of PG synthesis rather than a decreased rate of degradation. IGF-1 had no effect on the molecular weight of synthesized PG nor on the nature and distribution of the constitutive glycosaminoglycan chains, both in medium and in cell layer. The stimulation of PG synthesis by IGF-1 appeared to be due, at least partially, to an increase of glycosylation processes. IGF-1 effect was mediated by the classical tyrosine kinase signalling process, since IGF-1 action on PG synthesis was abolished by genistein and tyrphostin A9, two well known tyrosine kinase inhibitors. The increase of PG synthesis was accompanied with an undersulfation of constitutive glycosaminoglycan (GAG) chains (chondroitin sulfate and heparan sulfate chains) since the [35S]/[3H] ratio was reduced by about 20-25% in presence of IGF-1. Although the mechanism of hyaluronic acid synthesis was completely different from those of other GAG, IGF-1 also dramatically enhanced its production by PT cells. PMID- 9332450 TI - Prostaglandin E2 enhances type 2-bradykinin receptor expression in human glomerular podocytes. AB - We examined the effect of prostaglandin E2 (PGE2) on bradykinin (BK) binding, BK dependent intracellular calcium and inositol phosphate (i.p.) concentrations and BK mRNA in human glomerular visceral epithelial cells (hGVEC). PGE2 (10 nM) produced a concentration-dependent increase in [3H]-BK specific binding after a lag time of 24 h with a threshold at 0.1 nM. This increase appeared to be mediated exclusively by an increase in BK receptor (BKR)-2 expression. Scatchard analysis of [3H]-BK saturation binding showed that PGE2 produced an increase in the receptor site density without a change in the apparent dissociation constant. PGE2 also markedly stimulated cAMP production. This effect was thought to mediate the increase in expression of BKR-2 as 8-bromo cAMP and various cAMP-stimulating agents acted similarly. PGE2 did not change the BK-dependent intracellular IP3 and cytosolic calcium increases. The overexpression of BKR-2 in the presence of PGE2 was associated with an increase in mRNA as shown by the nuclease protection assay without any change in mRNA half-life. Cycloheximide, an inhibitor of protein synthesis, enhanced BKR-2 mRNA expression. In conclusion, treatment with PGE2 stimulates the synthesis of BKR-2 in hGVEC, possibly by interfering with an inhibitory protein involved in BKR-2 transcription. PMID- 9332451 TI - The 3' untranslated region plays a role in the targeting of metallothionein-I mRNA to the perinuclear cytoplasm and cytoskeletal-bound polysomes. AB - The mechanism of localisation of metallothionein-I (MT-I) mRNA was studied in transfected cells by in situ hybridisation and cell fractionation. Hepatoma cells were transfected with the 5'-untranslated region and coding region of the beta globin gene alone or linked to either the beta-globin 3'-untranslated region (3' UTR) or the MT-I 3'-UTR. The wild-type beta-globin mRNA and the beta-globin mRNA lacking its native 3'-UTR were present in free and cytoskeletal-bound polysomes to a similar extent and showed no localisation. Chimaeric globin-metallothionein transcripts were significantly enriched in cytoskeletal-bound polysomes and were localised in the perinuclear cytoplasm. Chimaeric globin-metallothionein and wild type globin transcripts were of similar stability. Chinese Hamster Ovary cells were transfected with constructs in which the MT-I 5'-untranslated region and coding sequences were linked to either the endogenous 3'-UTR or the glutathione peroxidase 3'-UTR. Wild-type MT-I transcripts were localised in the perinuclear cytoplasm but the chimaeric MT-I-glutathione peroxidase transcripts showed no distinct localisation. The results indicate that the 3'-UTR of MT-I mRNA contains a localisation signal which promotes both the association of the mRNA with the cytoskeleton and its perinuclear localisation. PMID- 9332452 TI - Two pathways for insulin metabolism in adipocytes. AB - Using selected conditions, the appropriate collagenase, albumin and cell treatment, a preparation of isolated adipocytes was developed with no extracellular insulin degrading activity. Cell mediated insulin degradation rates were 0.68% +/- 0.05%/100,000 cell/h using trichloracetic acid precipitability as a measure. Chloroquine (CQ) increased cell-associated radioactivity and decreased degradation while dansylcadaverine (DC), PCMBS and bacitracin (BAC) decreased degradation with no effect on binding. Extraction and chromatography of the cell associated radioactivity showed 3 peaks, a large molecular weight peak, a small molecular weight peak and an insulin-sized peak. CQ, DC and BAC all decreased the small molecular weight peak while CQ and DC also increased the peak of large molecular weight radioactivity. Cell mediated insulin degradation in the presence of combinations of inhibitors suggested two pathways in adipocytes, one affected by inhibitors of the insulin degrading enzyme (IDE) (bacitracin and PCMBS) and the other altered by cell processing inhibitors (DC, CQ and phenylarsenoxide). Chloroquine altered the pattern of the insulin-sized cell-associated HPLC assayed degradation products, further supporting two pathways of degradation; one a chloroquine-sensitive and one a chloroquine-insensitive pathway. PMID- 9332453 TI - Developmental changes in regulation of the Na+, K(+)-ATPase alpha 3 isoform by thyroid hormone in ferret heart. AB - Ferret heart expresses the alpha 1- as well as the alpha 3-isoform of the Na+, K(+)-ATPase. We have shown previously that the alpha 3 isoform is differentially upregulated during postnatal cardiac development and that in adult ferrets expression of alpha 3 is not responsive to regulation by thyroid hormone (TH). Since developmental-stage dependent effects of TH have been reported previously, the present study examined whether effects of TH on expression of the Na+, K(+) ATPase isoforms in ferret heart is modulated during development and possible mechanisms were examined. Ferrets of different age groups were treated with TH and the relative abundance of Na+, K(+)-ATPase isoforms in ferret myocardium was determined by immunoblotting. Thyroid hormone (T3; 50 micrograms/100 g body weight on 3 alternating days, s.c.) increased protein levels of the alpha 3 isoform, but not that of alpha 1 or beta 1, in myocardium of 5-day-old and 3-week old ferrets. By contrast, in myocardium of 6- and 8-week-old ferrets T3 failed to increase protein levels of alpha 1 and alpha 3. To determine whether elevated plasma levels of TH during development plays a role in the transition, mature ferrets were first made hypothyroid before TH treatment. In these hypothyroid ferrets expression of the alpha 3 isoform remained unresponsive to TH (T4, 0.5 mg/kg for 7 days, s.c.). The transition from TH-responsive to TH-unresponsive appears to be isoform-specific because in skeletal muscle of 8-week-old ferrets and in hypothyroid ferrets the alpha 2 isoform is upregulated by TH. Finally, there appears to be functional thyroid hormone receptors throughout development because in each age group TH effectively induced expression of alpha-MHC in the myocardium. In conclusion, these findings demonstrate that expression of alpha 3 isoform in the myocardium of newborn ferret is responsive to TH; however, the responsiveness terminates between 3- and 6-weeks of age. Neither elevated endogenous TH level nor a lack of functional thyroid hormone receptor appears to be responsible for the transition from TH-responsive to TH-unresponsive. PMID- 9332454 TI - Calpain and calpastatin in myoblast differentiation and fusion: effects of inhibitors. AB - Myoblast differentiation and fusion to multinucleated muscle cells can be studied in myoblasts grown in culture. Calpain (Ca(2+)-activated thiol protease) induced proteolysis has been suggested to play a role in myoblast fusion. We previously showed that calpastatin (the endogenous inhibitor of calpain) plays a role in cell membrane fusion. Using the red cell as a model, we found that red cell fusion required calpain activation and that fusibility depended on the ratio of cell calpain to calpastatin. We found recently that calpastatin diminishes markedly in myoblasts during myoblast differentiation just prior to the start of fusion, allowing calpain activation at that stage; calpastatin reappears at a later stage (myotube formation). In the present study, the myoblast fusion inhibitors TGF-beta, EGTA and calpeptin (an inhibitor of cysteine proteases) were used to probe the relation of calpastatin to myoblast fusion. Rat L8 myoblasts were induced to differentiate and fuse in serum-poor medium containing insulin. TGF-beta and EGTA prevented the diminution of calpastatin. Calpeptin inhibited fusion without preventing diminution of calpastatin, by inhibiting calpain activity directly. Protein levels of mu-calpain and m-calpain did not change significantly in fusing myoblasts, nor in the inhibited, non-fusing myoblasts. The results indicate that calpastatin level is modulated by certain growth and differentiation factors and that its continuous presence results in the inhibition of myoblast fusion. PMID- 9332455 TI - CCK causes rapid tyrosine phosphorylation of p125FAK focal adhesion kinase and paxillin in rat pancreatic acini. AB - Recent studies show CCK stimulates tyrosine phosphorylation (TYR PHOSP) of a number of proteins and evidence from the pancreas and other cellular systems suggest this could be important in mediating some of CCK's growth and secretory effects. In other tissues various neuropeptides such as bombesin can cause tyrosine phosphorylation of p125 focal adhesion kinase (p125FAK) and paxillin which are important in mediating their growth effects. The purpose of the present study was to determine the effects of CCK in rat pancreatic acini on the TYR PHOSP of these latter proteins. In dispersed rat pancreatic acini, cell lysates were incubated with an anti-phosphotyrosine mAb (PY20) which was immunoprecipitated and then analyzed by Western blotting with anti phosphotyrosine mAb (4G10), anti-p125FAK mAb or anti-paxillin mAb. CCK-8 at 5 min increased TYR PHOSP of five proteins of molecular weight > 60,000 including a broad M(r) 110-130,000 and M(r) 70-80,000. An increase in TYR PHOSP of both p125FAK and paxillin was detected within 1 min of adding CCK and reached a maximum at 2.5 min with a 9.1 +/- 1.9-fold increase for p125FAK and 3.6 +/- 0.6 fold for paxillin. CCK-8 caused a half-maximal increase in TYR PHOSP of p125FAK at 0.1 nM and paxillin at 0.03 nM. CCK-JMV also stimulated an increase in TYR PHOSP of both proteins, but was only 50% as efficacious as CCK-8. CCK-JMV caused a half-maximal increase at 10 nM and maximal at 1 microM for both proteins. To investigate whether the low affinity CCK receptor state also caused TYR PHOSP of both proteins, increasing concentrations of CCK-JMV were added to a maximally effective CCK-8 concentration (1 nM). Detectable inhibition of CCK-8-stimulated TYR PHOSP occurred with 1 microM CCK-JMV and with 3 microM CCK-JMV the CCK-8 stimulated response was inhibited 50% and was the same as that seen with CCK-JMV alone. These studies demonstrate that in rat pancreatic acini, CCK causes rapid TYR PHOSP of both p125FAK and paxillin. This stimulation is mediated by both the high affinity and low affinity CCK receptor states. This phosphorylation of these proteins could be important in mediating CCK's effect on the cytoskeleton or growth effects as shown for a number of other agents (oncogenes, neuropeptides, integrins). PMID- 9332456 TI - Photo-immobilization of epidermal growth factor enhances its mitogenic effect by artificial juxtacrine signaling. AB - Photo-reactive epidermal growth factor (EGF) was synthesized by coupling EGF with azidobenzoic acid and was immobilized onto the wells of a polystyrene culture plate by photo-irradiation. The photo-immobilized EGF enhanced the growth of anchorage-dependent cells more than native or azidobenzoyl derivatized EGF. A small amount of photo-immobilized EGF was sufficient to enhance the growth of cells and the maximal mitogenic effect was greater than that of native or derivatized EGF. On the other hand, the photo-immobilized EGF did not enhance growth of anchorage-independent cells. In addition, signal transduction in the cells adhered only on the EGF-immobilized surface was observed by staining of phosphotyrosine residues by anti-phosphotyrosine antibodies. These results showed that the enhanced cell growth was due to direct interaction between the cells and the immobilized EGF. Photo-immobilization could be a universal means of fixing growth factors onto an artificial matrix that is devoid of chemically functional groups scaffolding growth factors and could provide a new tool to elucidate signal transduction mechanism and could lead to the development of a new protein free cell culture system or tissue engineering materials. PMID- 9332457 TI - Effect of rapamycin on prolactin-stimulated S6 kinase activity and milk product formation in mouse mammary explants. AB - The in vitro effects of rapamycin on prolactin (PRL)-stimulated S6 kinase activity and milk product synthesis were investigated in cultured mouse mammary tissues. Mouse mammary gland explants were initially incubated for 24 h in M199 media containing 1 microgram/ml insulin and 10(-7) M cortisol. A subsequent treatment of the tissues with 1 microgram/ml PRL for 12 h caused a 98% increase in S6 kinase activity in the cytosolic fraction; effects were not observed at earlier times. PRL at or above 500 ng/ml was needed to elicit a maximum stimulation of S6 kinase activity, and this response was specific for lactogenic hormones (PRL, hPL, hGH). Rapamycin, an inhibitor of 70 K S6 kinase, was employed to assess the possible physiological role of S6 kinase in the PRL stimulation of milk product formation. Rapamycin (25-100 ng/ml) impeded, in a dose-dependent fashion, the PRL stimulation of casein, lipid and lactose synthesis in concert with its inhibition of cytoplasmic S6 kinase activity. These results suggest a possible role for the activation of 70 K kinase in the signalling pathway for the PRL regulation of milk product synthesis in the mammary gland. PMID- 9332459 TI - Bromodeoxyuridine improves the cytotoxic effect of cisplatin: a comparison with 5 fluorouracil. AB - We compared the effects of the radiosensitizers, 5-bromo-2'-deoxyuridine (BUdR) and 5-fluorouracil (5-FU) alone and in combination and cis diamminedichloroplatinum (cisplatin, DDP) on the growth of B16 amelanotic melanoma (B16a) tumors in mice. In a preliminary study, tumor growth was significantly inhibited in the presence of BUdR and was further reduced with the combination of BudR and DDP. In a second experiment, BUdR was found to be more effective than 5-FU when used in combination with DDP. At the completion of the study, tumor volumes as a percentage of control values in mice treated with a single drug were as follows: 5-FU (50 mg/kg per day for 7 days) 76.5% (P < 0.05), BUdR (100 mg/kg per day for 7 days) 68% (P < 0.05) and DDP (5 mg/kg x 3) 54% (P < 0.01). Combining 5-FU and DDP at these dosages reduced volumes to 38% (P < 0.01), while BUdR + DDP-treated mice had tumor volumes only 28% (P < 0.001) the size of untreated controls. Furthermore, the toxicity, as demonstrated by a decrease in body weight and an increase in mortality, was more severe in mice receiving 5-FU than in those receiving in BUdR. DDP interacts synergistically with either BUdR or 5-FU in its cytotoxic action in vivo. No such relationship could be demonstrated in vitro, suggesting that the pharmacologic activity of these drugs may be responsible for the antitumor activity than direct cytotoxic effects. We propose that BUdR is more effective than 5-FU as a potentiator of DDP in this murine melanoma model. PMID- 9332458 TI - Pharmacokinetics of adriamycin and cisplatin for anhepatic chemotherapy during liver transplantation. AB - We investigated the pharmacokinetics of cytotoxic anticancer agents administered under anhepatic conditions. Beagle dogs underwent either a sham operation consisting of laparotomy only (control group, n = 11) or a laparotomy and total hepatectomy under venovenous bypass (anhepatic group, n = 12). Each dog received a bolus intravenous injection of either Adriamycin (1 mg/kg) or cisplatin (1 mg/kg). The plasma and urine concentrations of each drug were measured at intervals for up to 2 h after drug injection. The dogs given Adriamycin were then sacrificed to determine tissue drug concentrations in the liver (controls only), spleen, kidney, heart, lung, skeletal muscle and small intestine. The control and anhepatic groups showed similar Adriamycin profiles during the initial 5 min after drug injection. However, subsequently, the plasma Adriamycin concentrations remained persistently higher in the anhepatic dogs than in the controls, yielding a two-fold elevation of the mean area under the concentration-time curve in the anhepatic group (P < 0.01 vs controls). The renal clearance values did not significantly differ between the two groups. The tissue Adriamycin concentrations in all measured organs, excluding the liver, were higher in the anhepatic group than in the controls. In a second set of experiments with cisplatin, the plasma platinum concentrations did not significantly differ between the two groups throughout the time course. However, the renal clearance of platinum in the anhepatic dogs showed a fourfold increase compared with that in the controls (P < 0.01). These pharmacokinetic data suggest that Adriamycin carries the risk of increased systemic toxicities, while cisplatin may be associated with increased renal toxicity when administered during the anhepatic period of liver transplantation. PMID- 9332460 TI - Cisplatin-based combination chemotherapy for elderly patients with non-small-cell lung cancer. AB - PURPOSE: To compare the response rates, toxicities and survival durations of elderly patients (70 years of age or more) with those of younger patients (less than 70 years of age) with non-small-cell lung cancer (NSCLC) treated with cisplatin-based chemotherapy. PATIENTS AND METHODS: We analyzed retrospectively the data of 203 assessable patients entered on a prospective randomized trial of cisplatin-based combination chemotherapy. Chemotherapy consisted of three dosage regimens: (1) vindesine and cisplatin (VP); (2) mitomycin, vindesine and cisplatin (MVP); or (3) etoposide and cisplatin alternating with vindesine and mitomycin (EP/VM). RESULTS: A greater proportion of elderly patients had localized disease and more squamous cell carcinoma than non-elderly patients. The overall response rates were 44% in the elderly group and 28% in the non-elderly group. In the EP/VM arm, the response rate was significantly better in the elderly group than in the non-elderly group. The frequency of grade 4 leukocytopenia in the MVP and EP/VM arms in the elderly group was significantly greater than in the non-elderly group (P < 0.05). No differences were found in nonhematological toxicities between the two groups. There was no difference in overall survival between the groups. CONCLUSION: Elderly patients treated with mitomycin-containing regimens have higher hematologic toxicities than younger patients. The results of this study are consistent with the previously reported pharmacologic data on mitomycin suggesting altered pharmacokinetics in elderly patients. The improved response rate in the elderly patients was probably because more elderly patients had earlier disease, squamous cell carcinoma and better performance status. Cisplatin-based chemotherapy was tolerable for most elderly NSCLC patients with good performance status. PMID- 9332462 TI - Multicentre CRC phase II trial of temozolomide in recurrent or progressive high grade glioma. AB - PURPOSE: Patients with progressive or recurrent supratentorial high-grade gliomas were entered into a multicentre phase II trial to evaluate the efficacy and toxicity of temozolomide. METHODS: The treatment schedule was 150-200 mg/m2 per day orally for 5 days repeated every 28 days. Response evaluation was by a combination of neurological status evaluation (MRC scale) and imaging. RESULTS: Of 103 eligible patients enrolled, 11 (11%) achieved an objective response and a further 48 (47%) had stable disease. The median response duration was 4.6 months. Response rates were similar for anaplastic astrocytomas (grade III) and glioblastoma multiforme (grade IV) tumours. Predictable myelosuppression was the major toxicity. CONCLUSIONS: The observation of objective responses and tolerable side effects in this heterogeneous population of patients supports the further investigation of this agent in high-grade gliomas. PMID- 9332463 TI - Methyl-beta-cyclodextrin in HL-60 parental and multidrug-resistant cancer cell lines: effect on the cytotoxic activity and intracellular accumulation of doxorubicin. AB - The purpose of this work was to determine the role of methyl-beta-cyclodextrin (MEBCD) in combination with doxorubicin (DOX) on the cellular proliferation of a sensitive parental and a multidrug-resistant human cancer cell line (HL-60 S and HL-60 R) and to study the effect of MEBCD on DOX intracellular accumulation. The cytotoxicity of DOX at five concentrations (50-50,000 nM) was evaluated with or without the coadministration of four fixed noncytotoxic concentrations of MEBCD (100, 200, 500, and 1,000 microM). Intracellular DOX concentrations were determined by a high-performance liquid chromatography (HPLC) method with fluorescence detection. MEBCD applied at 500 and 1000 microM in combination with doxorubicin (DOX) significantly potentiated the activity of DOX used alone on both sensitive and multidrug-resistant cell lines; 50% growth-inhibitory (IC50) ratios (IC50 MEBCD-DOX/IC50 DOX) were about 3:4 and 1.6:4 for HL-60 S and HL-60 R, respectively. Moreover, intracellular DOX accumulation, determined by HPLC during 6 h of drug exposure, was about 2-4 times higher for cells treated with MEBCD in combination with DOX than in those treated with DOX alone. Similar results were obtained using other paired MCF 7 sensitive and resistant cell lines. Correlation between these results and an MEBCD-cell membrane interaction was discussed. These initial data provide a basis for the potential therapeutic application of MEMBCD in cancer therapy. PMID- 9332461 TI - Sensitivity to camptothecin of human breast carcinoma and normal endothelial cells. AB - PURPOSE: To assess parameters that might determine resistance to the topoisomerase I inhibitor, camptothecin (CPT), the sensitivities of three established human breast cancer cell lines (ER-) and of normal bovine endothelial cells to CPT in the free form and incorporated into liposomes (LCPT), were contrasted with topoisomerase I (topo I) content and activity, and with cell cycle response to CPT treatment. METHODS: Drug sensitivities were determined using the tetrazolium dye assay and by 3H-thymidine incorporation. Topo I levels were determined by Western blot analysis, and catalytic activity was determined with a plasmid relaxation assay, using nuclear protein from each cell line. CPT stabilization of cleavable complexes in nuclear extracts was determined using a labeled oligonucleotide with a specific topo I cleavage site. Cell cycle response to CPT was determined by flow cytometric analysis of propidium iodide-stained nuclei. RESULTS: CPT was extremely potent against MDA-MB-157 cells with an IC50 value of 7 nM compared with IC50 values of 150 nM for GI 101A and 250 nM for MDA MB-231 cells. In contrast, CPT inhibited the incorporation of 3H-thymidine at very low doses in GI 101A and MDA-MB-231 cells with IC50 values of 9 nM and 5 nM, respectively; while MDA-MB-157 cells did not stop incorporating 3H-thymidine until very high doses (500 nM) of CPT were used. When incorporated into multilamellar liposomes (LCPT), CPT retained its potency, with IC50 values similar to that of the free drug. No correlation was found between CPT-induced cytotoxicity and any of the topo I parameters determined. Cell cycle analysis, however, showed an accumulation of cells in G2/M phase after 24 h treatment with low doses (5 nM) of CPT in only GI 101A and MDA-MB-231 cells with no arrest in normal endothelial or MDA-MB-157 cells. At higher doses (50 nM), however, a dramatic accumulation of cells in the S phase was observed in MDA-MB-157, MDA-MB 231 and GI 101A cells. In contrast, a G2/M phase block was seen with the normal bovine endothelial cells using the higher doses of CPT. CONCLUSIONS: The results suggest that cell cycle regulation plays an important role in determining the effect of CPT on malignant and normal cells. The possible mechanisms explaining the sensitivities of the two cellular compartments to the action of CPT are discussed. PMID- 9332464 TI - Combined action of paclitaxel and cisplatin against wildtype and resistant human ovarian carcinoma cells. AB - PURPOSE: The combination of paclitaxel (PTX) and cisplatin (DDP) shows good clinical efficacy against ovarian cancer. In order to examine the potential cellular basis for this, and provide leads as to how to optimize the combination, we examined the role of sequence of exposure to PTX and DDP on cell growth in vitro. METHODS: Four human ovarian carcinoma cell lines, A121, A2780/WT, A2780/DX5B and A2780/CP3, two human head and neck carcinoma cell lines, A253 and FaDu, and the human ileocecal carcinoma cell line, HCT-8, were treated with PTX + DDP with seven schedules: (A) 96 h exposure to PTX + DDP; (B) 24 h PTX alone, then 72 h PTX + DDP; (C) 4 h DDP alone, then 92 h PTX + DDP; (D) 24 h PTX alone, 4 h DDP alone, then 68 h drug-free; (E) 4 h DDP alone, 24 h PTX alone, then 68 h drug-free; (F) 3 h PTX alone, 1 h DDP alone, then 92 h drug-free; and (G) 1 h DDP alone, 3 h PTX alone, then 92 h drug-free. Each of 66 two-drug experiments included five plates (440 randomly treated wells per experiment). Cell growth was measured by the sulforhodamine B assay. The nature and the intensity of the drug interactions were assessed by fitting a seven-parameter model to data with weighted nonlinear regression, enabling the estimation of an interaction parameter, alpha, with its standard error. RESULTS: Overall there was very little departure from Loewe additivity: 43 experiments showed Loewe additivity, 10 showed Loewe antagonism, and 13 showed slight Loewe synergy. In vitro Loewe synergy was rare, was small when present, and reproducible only for the A121 and HCT-8 cells exposed to schedule D (24 h PTX prior to 4 h DDP). Isobolographic analysis showed complex combined-action surfaces with regions of local Loewe synergy and antagonism. CONCLUSION: It appears unlikely that the good clinical efficacy of the combination is primarily caused by a synergistic interaction at the cellular level. PMID- 9332466 TI - GS-164, a small synthetic compound, stimulates tubulin polymerization by a similar mechanism to that of Taxol. AB - PURPOSE: During our search for new microtubule effectors as anticancer agents, we have found that a small synthetic molecule designated GS-164 interferes with the assembly of porcine microtubule proteins and has cytotoxic activity against a wide range of human tumor cell lines. In this study, we investigated mode of action of the compound in comparison with Taxol and colcemid. METHODS: To gain an insight into the mode of action of GS-164, we used an in vitro microtubule polymerization assay and a flow-cytometric measurement technique. Microtubule organization and the level of tubulin polymerization in HeLa cells were also examined by immunofluorescence microscopy and cytoskeletal protein analyses, respectively. RESULTS: GS-164 stimulated assembly of microtubule proteins in vitro in a concentration-dependent and a GTP-independent manner. Furthermore, as with Taxol, the microtubule polymerization induced by GS-164 was antagonized by podophyllotoxin, a tubulin polymerization inhibitor, and microtubules formed by GS-164 were resistant to disassembly by calcium or low temperatures. GS-164 in the micromolar range arrested the cell cycle of HeLa cells in the mitotic phase leading to cell death. GS-164 also increased the amounts of cellular microtubules in HeLa cells, resulting in the formation of microtubule bundles. CONCLUSION: These results indicate that GS-164 stimulates microtubule assembly by a similar mechanism to that of Taxol. A comparative conformational analysis of GS-164 and Taxol suggested that the structure of the former mimics the minimum essential sites of Taxol required to exert the Taxol-like activities described above. Although the cytotoxicity of GS-164 against human tumor cells was 1000-fold lower than that of Taxol and GS-164 was one-tenth as active as Taxol in vitro, these findings pave the way for synthesizing clinically useful anticancer agents using GS-164 as a lead compound. PMID- 9332465 TI - The inactivation of doxorubicin by long ultraviolet light. AB - PURPOSE: Irradiation of doxorubicin (DOX) dissolved in RPMI medium 1640 by long ultraviolet (UVA) light results in a rapid decline in the cytotoxic activity of the drug. The present study was designed to sort out which component(s) of this medium are associated with the UVA inactivation of DOX. METHODS: The effects of UVA irradiation of DOX in solutions of various compositions were evaluated by measuring the changes in the drug growth inhibitory activity in P388 cells and in the DOX absorbance spectrum. RESULTS: Riboflavin seemed to be the major photosensitizing component in the medium and the effect was enhanced by the presence of histidine, methionine, tryptophan and tyrosine but not by other amino acids. The changes in DOX resulting from UVA irradiation in the presence of riboflavin, were not blocked by 1,4-diazabicyclo [2.2.2]octane (5 mM), superoxide dismutase (300 units/ml), catalase (150 units/ml) or sodium benzoate (50 mM). The effects of UVA light on doxorubicin could be prevented by excess ascorbic acid. CONCLUSIONS: The effects of UVA on DOX are mediated by riboflavin. The photoexcited riboflavin apparently interacts directly with DOX rather than by first forming reactive oxygen species. The results suggest that the photoinactivation of DOX may involve an oxidation step. The mechanism by which certain amino acids facilitate the photoinactivation of DOX is not known. It is suggested that patient intake of riboflavin and exposure to the sun and fluorescent light could affect the outcome of anthracycline treatment. PMID- 9332467 TI - The effects of anesthesia on the biodistribution of drugs in rats: a carboplatin study. AB - PURPOSE: Anesthetics can alter the biodistribution profile of drugs and, consequently, the regional pharmacokinetics of antineoplastic drugs at the tumor site. The effect of coadministered anesthetics on the biodistribution profile of carboplatin was studied in rats. METHODS: Female Wistar rats were used to compare the effects of ketamine/xylazine, thiopental and pentobarbital on the biodistribution of 30 mg/kg radiolabelled 195mPt-carboplatin administered intravenously, with conscious rats as the control group. Blood and urine samples were collected between 5 and 120 min. RESULTS: The percentage values of the injected dose of platinum per ml (%ID/ml) in plasma at the final time-point were respectively, 0.557%, 0.156%, 0.115% and 0.086%, in pentobarbital-, ketamine/xylazine- and thiopental-injected rats, and in conscious animals. Following the same sequence of groups, the %ID/ml values of platinum in the cumulative urine were 0.001%, 0.619%, 0.184% and 0.118%, respectively. Urine output varied from very little in the pentobarbital group, to several milliliters in the other groups. CONCLUSIONS: There was an increase of almost 100-fold in total platinum uptake in the kidneys, cerebrum and cerebellum of rats receiving pentobarbital over the uptake in the control rats, whereas the biodistribution profile of the thiopental group had the least variance. These results demonstrate the importance of anesthetic selection in animal pharmacokinetic studies, as it influences the biodistribution and pharmacokinetic profile of the drug being studied. PMID- 9332468 TI - Intermittent hepatic arterial infusion of high-dose 5FU on a weekly schedule for liver metastases from colorectal cancer. AB - PURPOSE: We performed phase I and II studies to examine the usefulness of intermittent hepatic arterial infusion of high-dose 5-fluorouracid (5-FU) for patients with liver metastasis from colorectal cancer. METHODS: As the phase I study, 1000, 1250 and 1500 mg/m2 of 5-FU were administered over 5 h by hepatic arterial infusion on a weekly schedule to establish the recommended dose. Based on the results of the phase I study, the phase II study was performed to confirm the efficacy of the recommended dose thus obtained. RESULTS: In the phase I study, the dose-limiting factors of this therapy were gastrointestinal and central nervous system toxicities, and the recommended dose was judged to be 1000 mg/m2. In the phase II study, 1000 mg/m2 of 5-FU was administered over 5 h once a week on an outpatient basis, and this therapy was repeated as long as possible. The response rate was 78% (25/32), with an overall median survival time of 25.8 months (without extrahepatic lesions 25.9 months; with extrahepatic lesions 17.3 months). CONCLUSIONS: (1) Compared with conventional continuous infusion, the advantages of this therapy were that it caused no decrease in the patient's quality of life as a result of being permanently equipped with a pump and it thus enabled more cost-effective use of the pump, (2) The phase II study on 32 patients showed that this therapy caused no serious toxicities, with a response rate of 78% and a survival time of 25.8 months, which exceeded the results with conventional continuous infusion. If the reproducibility of these results is established in further studies involving multicenter collaboration, this therapy will be able to become the standard local chemotherapy for liver metastases from colorectal cancer. (3) Important problems remaining to be solved are improvement of the technical aspects and studies of combined use with systemic chemotherapy. Furthermore, to finally determine the position of this therapy in the treatment system for liver metastasis from colorectal cancer, it is necessary to conduct comparative trials versus systemic chemotherapy, using the survival time as the end-point. PMID- 9332469 TI - Phenytoin-induced alteration in the N-dechloroethylation of ifosfamide stereoisomers. AB - CASE: A suspected alteration in ifosfamide (IFF) metabolism and pharmacokinetics was observed in a pediatric patient receiving phenytoin. METHODS: Sequential plasma samples were obtained and analyzed for the concentrations of the enantiomers of IFF and their N-dechloroethylated metabolites (DCE-IFF) using a validated enantioselective gas chromatographic-mass spectrometric method. RESULTS: In the phenytoin-treated patient, the metabolic formation of IFF enantiomers was increased and the metabolic pattern of the N-dechloroethylation altered from non-phenytoin-treated patients: (R)-3-DCE IFF > > (S)-3-DCE-IFF = (S)-2-DCE-IFF > (R)-2-DCE-IFF (control) vs (S)-3-DCE-IFF = (S)-2-DCE-IFF > (R)-3 DCE-IFF > > (R)-2-DCE-IFF (patient). CONCLUSIONS: Previous studies have attributed the production of the (S)-2-DCE-IFF and (S)-3-DCE-IFF metabolites to the activity of CYP2B6 and (R)-2-DCE-IFF and (R)-3-DCE-IFF to the activity of CYP3A4. The results suggest that phenytoin induced the activity of CYP2B6 to a greater extent than CYP3A4. In addition, the patient, who was at least partially refractory to several other treatments, went into remission after IFF treatment suggesting that phenytoin pretreatment might increase IFF therapeutic efficacy. PMID- 9332470 TI - Radioenhancement by cisplatin with accelerated fractionated radiotherapy in a human tumour xenograft. AB - The aim of the present study was to investigate whether cisplatin would enhance the radioresponse of a human tumour xenograft when given in different schedules combined with accelerated fractionated radiation therapy. A human squamous carcinoma of the hypopharynx, FaDu, was grown in the thigh of athymic nude mice. Tumours were exposed to twice-daily 2-Gy fractions, applied 6 h apart over 2 weeks, 5 days a week, alone or combined with cisplatin given at maximally tolerated doses in three different schedules: (1) i.p. as a single bolus (SB) or (2) i.p. as a daily bolus at 30 min before the first daily radiation fraction or (3) s.c. as a continuous infusion through a mini-osmotic pump over 13 days, commencing 24 h prior to the first daily radiation fraction. The end point for the study was tumour growth delay (TGD), calculated as the difference between the delay in regrowth to 200% of the initial tumour size in treated versus control mice. SB cisplatin plus radiation showed only an additive effect on TGD, whereas daily-bolus and continuous-infusion cisplatin demonstrated a greater than additive effect when combined with accelerated fractionated radiation in this human tumour model. Cisplatin appears to be especially beneficial as a radiation enhancer when given throughout the course of radiation. PMID- 9332471 TI - Leptomeningeal metastasis from a paraspinal rhabdomyosarcoma with a der(13)t(1;13)(q23;q32) in a 14-month-old boy. AB - Cytogenetic analysis performed on a 14-month-old boy with a primary retroperitoneal/paraspinal alveolar rhabdomyosarcoma showed the presence of a der(13)t(1;13)(q23;q32) resulting in partial trisomy of the 1q23-->qter region and loss of the 13q32-->qter region. The present case is discussed with reference to a similar case reported in the literature. PMID- 9332472 TI - Hairy cell leukemia variant with t(2;8)(p12;q24) abnormality. AB - Hairy cell leukemia variant is an uncommon chronic B-cell lymphoproliferative disorder characterized clinically by splenomegaly and marked leukocytosis. Cytologically, the leukemic cells are distinguishable from those of classical hairy cell leukemia by the presence of single, central, and vesicular nucleoli. Cytogenetic information for this uncommon leukemia is scanty, although structural abnormalities involving 7q34 have been reported in few cases. We report a patient with hairy cell leukemia variant who has t(2;8)(p12;q34) but without [corrected] c-MYC oncogene rearrangement. PMID- 9332473 TI - Cytogenetic and fluorescence in situ hybridization analyses of a microcystic adnexal carcinoma with del(6)(q23q25). AB - Cytogenetic analysis of a case of microcystic adnexal carcinoma (MAC) revealed three unrelated clones with the karyotypes: 46,XX,del(6)(q23q25)[17]/46,XX,1 approximately 2dmin[4]/46,XX,t(1;3) (p10;q10)[2]. Analysis of the 6q- by fluorescence in situ hybridization (FISH) showed that it had resulted from a long arm deletion. The finding of a clonal 6q deletion as the sole karyotypic change in a MAC is of special interest because we previously have identified 6q deletions in different types of malignant salivary gland tumors. This observation, together with the histopathologic similarities between salivary gland tumors and sweat gland tumors, further emphasize the histogenetic relationships between these tumor types. PMID- 9332474 TI - Two different Philadelphia chromosomes in a cell line from an AML-M0 patient. AB - A second Philadelphia (Ph) chromosome is one of the most common nonrandom secondary chromosome changes in leukemias with 9;22 translocations. It has been suggested, and observed in two studies of masked t(9;22), that the second Ph chromosome is an exact duplication of the entire derivative chromosome 22. In a cytogenetic study of bone marrow cells from an acute myelogenous leukemia patient, a cell line carrying two different Ph chromosomes evidenced by a chromosome 22 centromeric heteromorphism was found. From this observation arose the question whether the second der(22) was a true Ph chromosome or whether it was a deleted chromosome derived from the normal chromosome 22 that did not contain the bcr-abl rearrangement. A fluorescent in situ hybridization (FISH) study with the t(9;22) probe revealed two bcr-abl positive signals on 60 of 100 interphase nuclei. The second Ph could have resulted from a mitotic crossing over; or, analogously to late-appearing Philadelphia chromosomes, it may be derived from a new chromatid translocation between the chromosomes 9 and 22 not involved in the initial t(9;22). PMID- 9332475 TI - Telomeric association in women with breast and uterine cervix cancer. AB - This study compares the frequency of telomeric associations in the peripheral blood of women suffering breast and cervix uterine cancer with a healthy control group. Two kinds of cultures were developed for each individual: with and without aphidicolin. In the normal cultures, the number of telomeric associations observed was 95.5 times higher in individuals affected by breast cancer and 41.3 times higher in those affected by cervix uterine cancer when compared to the control group (p < 0.001). In the cultures with aphidicolin, higher numbers of altered metaphases were observed in both groups as compared to the control groups (p < 0.001). Statistically significant differences (p < 0.001) could also be observed when comparing telomeric associations between the two types of cancer in both cultures. When we compared individuals affected by breast cancer in both types of cultures statistical differences were found (p < 0.05), and similar results were found in individuals affected by uterine cervix cancer (p < 0.001). The findings suggest that telomeric associations may be reflecting chromosome instability observed in cancer and that this instability behaves differently for various types of cancer. PMID- 9332477 TI - Chromosomal abnormalities in low grade chondrosarcoma and a review of the literature. AB - In this study, cytogenetic analysis of two low-grade chondrosarcomas revealed relatively simple chromosomal complements with structural rearrangements involving chromosomes 1, 6, and 12 [46,XY,add(16)(q24)[3]/46,XY,t(1;20)(q21;q11),t(6;17)(q23;q23)[3]/46,XY, t(4;14)(q12;q24),t(5;6)(q12;p21) [2] and 45,XY,t(12;16)(q13;q24), 14[17]/44,idem,add(4)(p16),-17,[2] respectively]. Previously published reports of chondrosarcoma have revealed structural abnormalities of chromosomes 1, 6, 9, 12, and 15 as common. Also, a correlation between the simplicity or complexity of the abnormalities seen and histologic grade has been suggested. The findings of the current study support these earlier observations. PMID- 9332476 TI - Increasing metastatic potential is associated with induced chromosome 14 translocations in a previously nonmetastatic murine melanoma cell line. AB - The purpose of these studies was to demonstrate causal effects of abnormalities induced in mouse chromosome 14 on tumorigenicity and metastasis using the K-1735 murine melanoma cell line. Because anomalies in chromosome 14 have previously been associated with increases in metastatic potential, we induced chromosome 14 anomalies in a nonmetastatic K-1735 clone 10 cells initially containing two normal copies of chromosome 14 by treatment with mitomycin C. Clone 10-M1, in which a small population of cells (approximately 4%) contained translocations involving chromosome 14, was isolated and injected into athymic nude mice. Unlike the parental C-10 cells, C-10 M1 cells produced experimental lung metastases. Chromosomal analysis of cell cultures established from both subcutaneous tumors and lung metastases indicated that approximately 35% of the cell population contained chromosome 14 anomalies suggesting that this chromosome may play a role in tumor growth and metastasis. PMID- 9332478 TI - Cytogenetic study of pituitary adenomas. AB - We report the results of cytogenetic studies on 23 pituitary adenoma specimens, using both the direct and short-term tissue culture methods. The direct method was applied to all of the specimens and allowed a karyotype to be identified in 15 of the processed samples (65%). Four tumors were shown to have a hypotriploid chromosomal constitution, two of which also presented structural clonal rearrangements: an isochromosome 1q,i(1)(q10) and a der(1)t(1;3)(p22;q21) were observed in two PRL-secreting adenomas, one of which also had a telomeric association involving the short arms of chromosomes 14 and 19. Telomeric associations of the long arms of chromosomes 11, 19, and 22 were observed in a near-diploid, non-secreting tumor showing monosomy 13. One other adenoma showed trisomies 8 and 12, a finding that was confirmed by means of the FISH analysis of chromosome 8 and 12 centromeric probes in the more than 300 scored nuclei. An apparently normal chromosome constitution was observed in the remaining nine cases. Short-term cultures were set up in 21 of the 23 samples, allowing us to obtain a karyotype in 18 specimens (85%). The six tumors that could not be analyzed using the direct method showed a normal karyotype. A diploid chromosome constitution was observed in the four tumors shown to be hypotriploid by the direct method as well as in the tumor with monosomy 13. The trisomies 8 and 12 identified by the direct method in one tumor were still observed, but a clone with a normal karyotype was also found. To the best of our knowledge, this is the only report of the results of cytogenetic studies on pituitary adenomas performed using both direct preparation and short-term culture. PMID- 9332479 TI - Low frequency of TEL/AML1 in adult acute lymphoblastic leukemia. AB - Translocation (12;21)(p13;q22) is a recently characterized aberration in acute lymphoblastic leukemia, and results in the fusion of the TEL and the AML1 genes. It is the most common translocation in pediatric acute lymphoblastic leukemia (ALL), occurring in about one third of the cases. To determine the frequency of TEL/AML1 in adult ALL, we studied 4 cases of T lineage ALL and 26 cases of B lineage ALL. Only one positive case was identified, giving a very low frequency of 3.3%. In this patient, TEL/AML1 was still detectable in complete hematologic remission. The apparent age predilection of t(12;21) warrants further investigations. PMID- 9332481 TI - Chromosomal anomalies in a case of proliferative myositis. AB - Proliferative myositis, a reactive lesion similar to proliferative fasciitis and nodular fasciitis, has only been cytogenetically described in one other report to date. This previously described case showed trisomy 2. Cytogenetic analysis and fluorescence in situ hybridization (FISH) of a proliferative myositis lesion in the present study did not reveal trisomy 2 but the following clonal translocation was observed: 46,XX,t(6;14)(q23;q32). PMID- 9332480 TI - A(870)E mutation of the androgen receptor gene in a patient with complete androgen insensitivity syndrome and Sertoli cell tumor. AB - In a 60-year-old woman with complete androgen insensitivity syndrome (CAIS) and Sertoli cell tumor, a germline mutation (A870E) in exon 8 of the androgen receptor (AR) gene could be detected. A sister of the patient was also affected by CAIS and developed a Sertoli cell tumor at age 56. The mutation has not been described so far and could be seen in a causal relationship with the development of this tumor. PMID- 9332491 TI - Dehydroepiandrosterone inhibits DNA synthesis of rat hepatocytes induced by partial hepatectomy or mitogen (ciprofibrate). AB - In a previous study we have shown that dehydroepiandrosterone (DHEA) inhibits hepatocyte DNA synthesis after short-term administration and induces hepatocellular carcinomas after long-term administration in the rat. It is not known whether DHEA is also capable of inhibiting replicative and mitogen-induced DNA synthesis. In the present study, we have evaluated the effect of DHEA on DNA synthesis in the rat liver after partial hepatectomy and mitogen administration. After partial hepatectomy, DHEA significantly inhibited DNA synthesis at 20, 26, 32 and 38 h. Similarly, combined administration of ciprofibrate, a peroxisome proliferator and mitogen, and DHEA also resulted in significant hepatocyte DNA synthesis. However, DHEA did not affect liver enlargement caused by ciprofibrate. This experimental system will serve as useful tool to evaluate the role of cell proliferation in carcinogenesis. PMID- 9332492 TI - Changes in the phosphorylation status of the 27 kDa heat shock protein (HSP27) associated with the modulation of growth and/or differentiation in MCF-7 cells. AB - We have used human mammary cells of the MCF-7 strain, which constitutively express high levels of the small heat shock protein HSP27 and we have compared the changes in the phosphorylation status of this protein together with changes in cell growth and/or morphology induced by the action of one of the following agents: (1) TPA (12-O-tetradecanoylphorbol-13-acetate), known as a differentiation inducer in MCF-7 cells; (2) OH-TAM (hydroxytamoxifen), which exerts a cytostatic and cytotoxic action; or (3) TNF alpha (tumour necrosis factor), which induces apoptotic cell death in this cell line. Our data show that TPA and TNF stimulate an immediate and massive phosphorylation of HSP27, whereas OH-TAM affect the phosphorylation status of the protein only after a 3 day delay. In the case of TPA, high levels of HSP27 phosphorylation were maintained for at least 4 days, along with growth inhibition and acquisition by the cells of a secretory phenotype. TPA and OH-TAM exerted similar immediated effects on cell growth, despite the different time course of their action on HSP27 phosphorylation. This excludes the possibility that the latter is a necessary consequence of, or an absolute requisite to, growth inhibition. With OH-TAM and TNF the increase in HSP27 phosphorylation was concomitant with the appearance of apoptosis, not observed with TPA. This indicates that increased phosphorylation of HSP27 is not specifically associated with the triggering or the execution of apoptosis in these cells. Altogether, our data support the concept that phosphorylated HSP27 is involved (and might then be rate limiting in some instances) in the execution of vital cell programmes (including resistance to stress, proliferation and differentiation), as well as in that of cell death. This is consistent with its role in actin polymerization and its position downstream of the p38/RK-type MAPkinase, itself a point of convergence for diverse signal transduction pathways. PMID- 9332493 TI - MIB-1 and S-phase cell fraction predict survival in non-Hodgkin's lymphomas. AB - The monoclonal antibody anti-Ki67 is used to detect proliferating cells, but its main limitation is the requirement of fresh-frozen material. On a series of patients with non-Hodgkin's lymphoma, we used a Ki67 equivalent monoclonal antibody, the recently proposed MIB-1, on formalin-fixed histopathological material using microwave antigen retrieval. MIB-1 expression was analysed in relation to other proliferation indices, such as autoradiographic H-thymidine labelling index (HTL1) and flow cytometric S-phase cell fraction (FCM-S) and to pathological status. Moreover, the prognostic relevance of the cell kinetic indices was defined in uni- and multivariate analyses including histology and tumour stage. The relationship between MIB-1 index and the other proliferation indices was statistically significant even though the correlation coefficient was around 0.6. The MIB-1 index was also related to the REAL (Revised European American Lymphoma) classification, but not to the Ann Arbor stage classification. Univariate analysis showed that the MIB-1 index was a significant predictor of 6 year survival in the overall series and in distinctly analysed low-grade and high grade lymphoma subgroups. With regard to S-phase indices, HTLI was a powerful prognosticator in patients with high-grade histologies and FCM-S in patients with low-grade histologies. Multivariate analyses revealed that MIB-1 index, HTLI and FCM-S retained their prognostic significance independent of histology. In conclusion, the MIB-1 antibody provides prognostic information in non-Hodgkin's lymphomas and has the main advantage that it can be used in formalin-fixed, paraffin-embedded specimens. PMID- 9332494 TI - Brief exposures of resting fibroblasts to okadaic acid stimulate DNA synthesis. AB - To study further the factors providing for cellular quiescence, we used okadaic acid (OA) at concentrations (0.1, 1, 10 or 100 nM) inhibiting type 1 and/or type 2A protein phosphatases in mammalian cell cultures. Brief (2 h) exposure of resting (0.2% serum for 72 h) NIH 3T3 mouse fibroblasts to OA with subsequent incubation of cells in a medium with 0.2% serum, stimulated DNA synthesis at all concentrations studied. Maximal stimulation was observed following pre-incubation of resting cells with 10 nM OA. Treatment of cycling cells (10% serum) with OA (2 h pulses at 12 h intervals for 72 h) prevented their exit to the resting state on transfer to a medium with 0.2% serum. Brief exposures of resting cells to OA did not affect the rate of protein synthesis. OA pulses in the late pre-replicative period had no effect on the entry of serum-stimulated cells into the S phase. Cell fusion experiments with resting (serum-deprived) and proliferating (serum stimulated) NIH 3T3 cells, using radioautography with a double-labelling technique, revealed that pre-incubation of resting cells with OA for 2 h before and after fusion abrogates their ability to suppress the onset of DNA synthesis in the nuclei of proliferating cells in heterodikaryons. The results indicate that protein phosphatases of type 1 and/or 2A may be involved in the growth arrest machinery that provides for cellular quiescence. PMID- 9332495 TI - Serum deprivation induces SV40 early promoter activity. AB - Proliferation and the expression of proliferation-associated genes are modulated by changing the serum concentration in the media of cultured cells. To determine if activity of the SV40 early promoter is modulated by serum, we examined the expression of SV40 early promoter driven marker genes in murine BALB/c 3T3T cells following serum deprivation or serum stimulation. SV40-promoter-regulated beta galactosidase and chloramphenicol acetyl transferase genes were studied following either transient or stable transfection. The results show that serum deprivation of growing cells induces SV40 promoter activity while serum stimulation of quiescent G0 cells suppresses it. Kinetic analyses show a significant induction of the SV40 promoter activity during the first 2 days of serum deprivation which is maintained at a high level for 15 days. The induction of reporter gene expression by serum deprivation was selective for the SV40 early promoter because such an effect was not observed using the Rous sarcoma viral promoter. Nuclear run-off assays further show that the transcription controlled by the SV40 early promoter is approximately twofold greater in cells rendered quiescent by serum deprivation for 72 h than in growing cells cultured in medium containing serum. These results suggest that one reason SV40 T transformed cells commonly fail to undergo quiescence following serum deprivation is that the SV40 promoter is induced. PMID- 9332496 TI - Default cycle phases determined after modifying discrete DNA sequences in plant cells. AB - After bromosubstituting DNA sequences replicated in the first, second, or third part of the S phase, in Allium cepa L. meristematic cells, radiation at 313 nm wavelength under anoxia allowed ascription of different sequences to both the positive and negative regulation of some cycle phase transitions. The present report shows that the radiation forced cells in late G1 phase to advance into S, while those in G2 remained in G2 and cells in prophase returned to G2 when both sets of sequences involved in the positive and negative controls were bromosubstituted and later irradiated. In this way, not only G2 but also the S phase behaved as cycle phases where cells accumulated by default when signals of different sign functionally cancelled out. The treatment did not halt the rates of replication or transcription of plant bromosubstituted DNA. The irradiation under hypoxia apparently prevents the binding of regulatory proteins to Br-DNA. PMID- 9332497 TI - Exogenous retinoic acid decreases in vivo and in vitro proliferative activity during the early migratory stage of neural crest cells. AB - We have previously demonstrated that directional migration of neural crest cells (NCC) is associated with a high cell density, resulting from an active cell proliferation. It is also known that treatment with retinoic acid (RA) causes a dose-dependent inhibition of proliferation of some cell types, and that administration of RA during the early stages of embryonic development, induces cranio-facial abnormal patterns corresponding to NCC derivatives. In view of these findings, it was of interest to determine if exogenous RA is a potential modulator of the mitotic rate of NCC, and to explore the hypothesis of an inhibitory effect exerted by RA on the proliferative behaviour of NCC in vivo and in vitro. Homogenates of RA-treated chick embryos showed a low [3H]dT incorporation, indicating a generalized diminution of DNA synthesis. The labelling index (LI = number of labelled cells/total number of cells) revealed that NCC from RA-treated and control embryos had higher values of [3H]dT incorporation than neural tube cells (P < 0.0001). Autoradiographs of RA-treated chick embryos showed a significantly lower [3H]dT incorporation in NCC at the prosencephalic and mesencephalic levels, as well as in the neural tube cells at the prosencephalic, mesencephalic and rhombencephalic levels, than in control chick embryos (P < 0.0001). NCC cultures treated with 1 or 10 microM RA had a significantly lower LI than in cultures treated with 0.1 microM RA or control cultures (P < 0.04). In chick embryos, the mitotic index of NCC was 0.026 for RA treated and 0.033 for controls, while the duration of the cell cycle was significantly longer in the NCC of RA-treated embryos (approximately 40 h) than in controls (approximately 25 h). The length of the cell cycle phases of NCC was similar in both experimental conditions, except for G1 phase, which was significantly longer in the RA-treated group than in controls. These results show that RA blocks DNA synthesis and lengthens the proliferative behaviour of NCC both in early chick embryos and in vitro, effects that could modify the morphogenetic patterns of NCC distribution through a decreased cell population. PMID- 9332498 TI - EGF effects on p53 in MDA-468 human breast cancer cells: implications for G1 arrest. AB - EGF, in pharmacological concentrations, inhibits cell proliferation of the MDA 468 human breast cancer cell line. Previously, we have demonstrated that this was characterized by a reversible cell cycle arrest at the G1-S boundary, concomitant with downregulation of mRNA levels for p53 (a point mutant, p53(273.His)). Since p53(273.His) is regarded as a gain-of-function mutant and acts to enhance cell proliferation, we hypothesized that the G1 arrest induced by EGF might be mediated by p53(273.His). In this study, we report an EGF-dependent altered conformation as indicated by immunofluorescence, while no significant immediate effects of EGF-treatment on p53(273.His) protein levels and synthesis were observed. These experiments demonstrated a decreased PAb 240 (mutant-specific) reactivity of nuclear p53(273.His) in EGF-treated cells, while that of PAb 1620 (wild-type specific) was enhanced. Staining with PAb 1801 (pan specific), on the other hand, showed little change upon EGF treatment. Further studies indicated a decreased phosphorylation of nuclear p53(273.His) in EGF-treated cells. These EGF dependent events were detected early enough to be attributed as causative of cell cycle arrest. We suggest that EGF-mediated, phosphorylation-dependent conformational change in nuclear p53(273.His), and in turn altered p53 function, may be responsible for EGF-dependent growth inhibition MDA-468 cells. PMID- 9332499 TI - Preferential oxidation of cardiac mitochondrial DNA following acute intoxication with doxorubicin. AB - The purpose of this investigation was to determine whether acute doxorubicin intoxication causes a preferential accumulation of 8-hydroxydeoxyguanosine (8OHdG) adducts to mitochondrial DNA (mtDNA) as opposed to nuclear DNA (nDNA), particularly in cardiac tissue. Adult male rats received a single i.p. bolus of doxorubicin (15 mg/kg) and were killed 1-14 days later. Acute intoxication with doxorubicin caused a 2-fold greater increase in 8OHdG adducts to mtDNA compared to nDNA, the concentration of adducts to both nDNA and mtDNA being 20%-40% greater for heart as opposed to liver. For both tissues, the relative abundance of adducts was highest at the earliest time-point examined (24 h) and decreased to control values by 2 weeks. The temporal dilution of 8OHdG adducts was not the result of cell hyperplasia and was only partially due to amplification of the mitochondrial genome, most probably via an increase in DNA copy number rather than a stimulation of mitochondrial biogenesis. PMID- 9332500 TI - Cytochrome bd terminal oxidase. PMID- 9332501 TI - The nucleotide regulatory sites on the mitochondrial KATP channel face the cytosol. AB - The mitochondrial KATP channel (mitoKATP) is richly endowed with regulatory sites for metabolites and drugs, but the topological location of these sites is unknown. Thus, it is not known whether ATP, GTP and acyl CoA esters regulate mitoKATP from the matrix or cytosolic side of the inner membrane, nor whether they all act from the same side. The experiments reported in this paper provide an unambiguous answer to these questions. Electrophysiological experiments in bilayer membranes containing purified mitoKATP showed that current is blocked asymmetrically by ATP. K+ flux experiments using proteoliposomes containing purified mitoKATP showed that mitoKATP is unipolar with respect to regulation by Mg2+, ATP, GTP, and palmitoyl CoA and that all of these ligands react on the same pole of the protein. This demonstration was made possible by the new finding that mitoKATP is 85-90% oriented inward or outward in liposomes, depending on the presence or absence of Mg2+ in the reconstitution buffer. K+ flux experiments in respiring rat liver mitochondria showed that mitoKATP was inhibited by palmitoyl CoA and activated by GTP when these agents were added to the external medium. Given that the inner membrane is impermeant to these ligands and that mitoKATP is unipolar with respect to nucleotide regulation, it follows that the regulatory sites on mitoKATP face the cytosol. PMID- 9332503 TI - Zinc, copper and iron and their interrelations in the growth of sickle cell patients. AB - In this study we evaluated the nutritional status of 34 children with sickle cell disease (SS). Results were compared to 9 siblings with sickle cell trait (AS) and 35 eutrophic children who presented normal hemoglobin and normal hemoglobin electrophoresis (AA). All of then came from low socioeconomic level. Analysis of the growth velocity curves revealed in SS group, tendency to increase deficit in weight and height with age. There was no relation between weight/height (W/H) and height/age (H/A) percentile and hemoglobin levels. There was no significant relation between nutritional status and severity of the disease. SS group showed significant skeletal maturation delay, the same did not occur with the siblings (AS group). Plasma zinc levels were significantly lower in SS group than in AS and AA groups. In SS group there was some association between lower plasma zinc levels and H/A percentile lower or equal to 10. Plasma copper levels were significantly greater in SS group than in AS and AA ones, and there was no relation between plasma copper levels and serum ferritin levels. In conclusion, our patients with sickle cell disease showed indexes of malnutrition, iron deficiency, hypercupremia and low plasma zinc levels related to low stature. PMID- 9332504 TI - Effects of dietary fat and nitrite on plasma and corporal density. AB - Two groups of nine weanling male rats each were fed different diets for 60 days. Group A (control) was fed a full casein diet containing 17% protein. The group B received the same diet plus nitrite, fried bacon and proline. Diet B induced increased body weight gain and increased plasma l-lactic acid and cholesterol levels, as well as a decrease in plasma selenium. We suggest that the adverse effects of diet B are related to peroxidation, with an increased nutritional need for selenium. PMID- 9332502 TI - Light-induced electrogenic events associated with proton uptake upon forming QB- in bacterial wild-type and mutant reaction centers. AB - Light-induced voltage changes (electrogenic events) were measured in wild-type and site-directed mutants of reaction centers (RCs) from Rhodobacter sphaeroides oriented in a lipid monolayer adsorbed to a Teflon film. A rapid increase in voltage associated with charge separation was followed by a slower increase attributed to proton transfer from solution to protonatable amino-acid residues in the vicinity of the QB site. In native reaction centers the proton-transfer voltage had a pH-dependent amplitude with two peaks at pH 4.5 and pH 9.7, respectively. In the Glu-L212-->Gln RCs the high-pH peak was absent, whereas in the Asp-L213-->Asn RCs the low-pH peak was absent and the high-pH peak was shifted to lower pH by about 1.3 pH units. The amplitudes of the electrogenic phases as a function of pH follow approximately the measured proton uptake from solution (P.H. McPherson, M.Y. Okamura, G. Feher, Biochim. Biophys. Acta, vol. 934, 1988, pp. 348-368) and are ascribed to proton transfer to amino acid residues upon QB- formation. The peak around pH 9.7 is ascribed to proton uptake predominantly by Glu-L212 and the peak around pH 4.5 to proton uptake predominantly by Asp-L213 or a residue strongly interacting with Asp-L213. PMID- 9332505 TI - Effect of germination on the protein content and on the level of specific activity of lipoxygenase-1 in seedlings of three soybean cultivars. AB - The effect of germination on the protein content and in the level of specific activity of lipoxygenase-1 on seedlings of Santa Rosa, FT-2 and Davis soybean cultivars were studied. Soybean seeds of the three cultivars were germinated from 0 to 72 hours in a seed germinator at 30 degrees C temperature, 100% relative humidity, and absence of light. The seeds were then frozen in liquid nitrogen, freeze-dried and milled. The lipoxygenase-1 was extracted with 0.02M thris-HCI buffer at pH = 8.2 and 13% saccharose for 30 minutes at 4 degrees C without shaking. The highest levels of specific activity of lipoxygenase-1 per milligram of flour (dry basis) in the Santa Rosa, FT-2 and Davis cultivars were observed between 18 and 36, 18 and 24, and 0 and 12 hours, respectively. The polinomial quadratic model best fits the regression analysis of the variable levels of specific lipoxygenase activity for cultivar Davis (R2 = 0.9197). Although for cultivars Santa Rosa (R2 = 0.9041)., and FT-2 (R2 = 0.7486) the 4th degree polynomial model has shown the best fit. The protein content in the three cultivars studied reached the maximum values at 48h germination. The germination induced a relative increase of the protein content but caused a reduction of the level of specific lipoxygenase-1 activity. PMID- 9332506 TI - Protection of travelers. PMID- 9332507 TI - Photo quiz. Cutaneous larva migrans. PMID- 9332508 TI - Resistance patterns of streptococcus pneumoniae from carriers attending day-care centers in southwestern Greece. AB - The resistance to beta-lactam and non-beta-lactam antibiotics of 133 nasopharyngeal isolates of Streptococcus pneumoniae recovered from December 1995 to February 1996 from children attending seven day-care centers in southwestern Greece was studied. Reduced susceptibility to one or more anti-microbial agents was found in 70 isolates (53%), as follows: penicillin, 17% intermediate, 12% resistant; cefotaxime, 10.5% intermediate, 1.5% resistant; trimethoprim sulfamethoxazole, 8% intermediate, 35% resistant; chloramphenicol, 27% resistant; tetracycline, 29% resistant; and erythromycin/clindamycin, 19% resistant. Eighty seven percent of penicillin-intermediate or -resistant strains belonged to serogroups/serotypes 19, 21, and 23. Fifty-six percent of the antibiotic resistant pneumococci were multiply resistant, including serogroup 6 strains that were penicillin-susceptible but resistant to all non-beta-lactam drugs tested, as well as serogroup 23 strains resistant to penicillin, chloramphenicol, tetracycline, and trimethoprim-sulfamethoxazole. The high incidence of antibiotic resistant pneumococci and the divergent and unique resistance patterns found in this study underline the need for global surveillance of S. pneumoniae to document the evolution and spread of resistant strains and to guide therapy. PMID- 9332509 TI - Prevalence of human immunodeficiency virus infection, mortality rate, and serogroup distribution among patients with pneumococcal bacteremia at Denver General Hospital, 1984-1994. AB - Pandemics of human immunodeficiency virus (HIV) type 1 infection and penicillin resistance highlight the urgency of preventing invasive pneumococcal disease with vaccination. We characterized pneumococcal serogroup distribution and the mortality rate among 460 patients with pneumococcal bacteremia from 1984 through 1994 at Denver General Hospital and the prevalence of HIV infection in patients for whom pneumococcal bacteremia was diagnosed from 1989 to 1994. Vaccine-related serogroups accounted for 426 isolates (92.6%), including 48 (92.3%) of 52 isolates from HIV-infected patients. Mortality among patients 15 years of age or older was higher during 1984-1988 (18[12.9%] of 140) than during 1989-1994 (10 [5.2%] of 191: rate ratio, 2.5; 95% confidence interval, 1.2-5.2). Of patients 15 59 years of age from 1989 to 1994, 44 (39.6%) of 111 men and three (7.3%) of 41 women were HIV-infected. Four (8.5%) of 47 HIV-infected patients and four (3.8%) of 105 other patients in this group died (age-weighted rate ratio, 1.8; 95% confidence interval, 0.5-6.2). We recommend routine screening of young adults with pneumococcal bacteremia for HIV infection and immunization of HIV-infected patients with pneumococcal vaccine (which includes most serogroups of infecting strains). PMID- 9332510 TI - Treatment of blastomycosis with higher doses of fluconazole. The National Institute of Allergy and Infectious Diseases Mycoses Study Group. AB - Recent clinical data suggest that fluconazole at daily doses of 200 to 400 mg for at least 6 months is moderately effective therapy for non-life-threatening blastomycosis. To examine the usefulness of higher doses of fluconazole therapy for this disorder, we conducted a multicenter, randomized, open-label study to determine the efficacy and safety of two different daily doses of fluconazole (400 and 800 mg) in the treatment of non-life-threatening blastomycosis. Of 39 patients evaluable for efficacy analysis, 34 (87%) were successfully treated, including 89% and 85% of patients who received 400 and 800 mg, respectively. Five (83%) of six patients for whom prior antifungal therapy had failed were successfully treated. The mean duration of therapy was 8.9 months for successfully treated patients. Nineteen patients (48%) reported adverse events, although most were minor. We conclude that fluconazole at daily doses of 400 to 800 mg for at least 6 months is effective therapy for non-life-threatening blastomycosis. PMID- 9332511 TI - Mycobacterium xenopi infection in patients with human immunodeficiency virus infection. AB - Mycobacterium xenopi is one of the most frequently isolated nontuberculous mycobacteria in Ontario, Canada. We reviewed the records of 28 human immunodeficiency virus (HIV)-infected patients from whom M. xenopi was isolated between 1982 and 1995. M. xenopi was recovered from respiratory specimens from 24 patients, most of whom had clinical and radiographic evidence of pulmonary disease. However, coexistent pulmonary infection due to other pathogens was found in 17 patients: Pneumocystis carinii (9 patients), cytomegalovirus (5), Haemophilus influenzae (2), Mycobacterium avium complex (2), Streptococcus pneumoniae (1), Staphylococcus aureus (1), Aspergillus species (1), and Histoplasma capsulatum (1). Three patients had bacteremia with M. xenopi, including two patients with pulmonary infection. Two of the bacteremic patients had chronic fever and a wasting syndrome. Twenty-one (75%) of the 28 patients were thought to be colonized, and seven patients (25%; of whom four had CD4 cell counts of < or = 50/mm3) were thought to have significant infection due to M. xenopi. Sixteen patients died, but in no case was death attributable to M. xenopi infection. In a region where M. xenopi is a relatively common mycobacterial isolate, the organism frequently colonizes HIV-infected patients. Significant disease occurs in those patients with more advanced HIV infection. PMID- 9332512 TI - Quadriplegia complicating Escherichia coli meningitis in a newborn infant: case report and review of 22 cases of spinal cord dysfunction in patients with acute bacterial meningitis. AB - We report a case of Escherichia coli meningitis complicated by spinal cord dysfunction in a neonate. This very rare complication of bacterial meningitis was probably caused by ischemia of the cord resulting from vasculitis. We review the 22 other reports of patients with this complication and discuss its pathogenesis. PMID- 9332513 TI - Pneumocystis carinii pneumonia in patients receiving chemotherapy for breast cancer. AB - Pneumocystis carinii pneumonia (PCP) is uncommon in patients undergoing chemotherapy for breast cancer. Most previously described patients with breast cancer and PCP received treatment with corticosteroids, a known risk factor for PCP. We describe two patients with metastatic breast cancer who developed PCP after receiving therapy with high doses of cyclophosphamide with peripheral blood stem cell support. Both patients developed fevers of unclear etiology in the setting of recovery of the neutrophil count. Only one patient had pulmonary symptoms. P. carinii infection was documented in both cases by bronchoscopy. One patient died after prolonged ventilatory support for PCP. Steroid exposure did not appear to be a risk factor for the development of PCP in either patient. Patients receiving sequential high doses of chemotherapy with stem cell support may be at increased risk for PCP. The role of prophylaxis for PCP in this setting should be continually redefined as the type and intensity of chemotherapy, as well as methods of procurement of autologous stem cells, continue to change. PMID- 9332514 TI - Pneumocystis carinii pneumonia in patients without human immunodeficiency virus infection. PMID- 9332515 TI - Pyogenic vertebral osteomyelitis of the posterior elements. AB - Lone pyogenic involvement of the posterior elements of the vertebrae is a rare event. We present two cases of isolated lumbar vertebral infection of the posterior elements and review 13 cases previously reported in the literature. Clinical presentation, laboratory and radiographic findings, microbiological etiology, and treatment were evaluated. Back pain, an elevated erythrocyte sedimentation rate, and computed tomography or magnetic resonance imaging were most useful in identifying the presence and extent of infection. Antibiotic therapy with or without surgical intervention was successful in all cases. PMID- 9332516 TI - Skin and soft-tissue manifestations of Shewanella putrefaciens infection. AB - Shewanella putrefaciens, a saprophytic gram-negative rod, is infrequently recovered from clinical specimens. Although a number of clinical syndromes have been attributed to S. putrefaciens, the pathogenic role of this agent remains largely undefined. We report 16 cases of S. putrefaciens infection that occurred at the Veterans General Hospital-Kaohsiung in Taiwan between 1990 and 1995. S. putrefaciens infection was associated with a wide clinical spectrum including bacteremia/septicemia, skin and soft-tissue infection, biliary tract infection, peritonitis, and empyema. Five of our patients had skin and soft-tissue manifestations, including fulminant periorbitofacial cellulitis, dacryocystitis, perineal abscess, finger abscess, and postcholecystectomy wound infection. These clinical features deviated from the chronic ulcers or infected burns of the lower extremities that have been described in previous reports. Seven (44%) of our 16 patients had bacteremia/septicemia, and all seven had underlying hepatobiliary diseases. S. putrefaciens was isolated in mixed cultures of specimens from 14 patients; Escherichia coli was the most common coisolate. Hepatobiliary diseases and malignancy were the major predisposing factors for S. putrefaciens infection of the biliary tract and S. putrefaciens bacteremia/septicemia. PMID- 9332517 TI - Effects of requiring prior authorization for selected antimicrobials: expenditures, susceptibilities, and clinical outcomes. AB - Antimicrobial control programs are widely used to decrease drug expenditures, but effects on antimicrobial resistance and outcomes for patients are unknown. When a requirement for prior authorization for selected parenteral antimicrobial agents was initiated at our urban, county teaching hospital, total parenteral antimicrobial expenditures decreased by 32%. Susceptibilities to all beta-lactam and quinolone antibiotics increased, with dramatic increased susceptibilities in isolates recovered in intensive care units, increased susceptibilities in isolates recovered in other inpatient sites, and little change in susceptibilities in isolates recovered in outpatient sites despite no change in infection control practices. For patients with bacteremia due to gram-negative organisms, overall survival did not change with restrictions. No differences occurred in the median time from initial positive blood culture to receipt of an appropriate antibiotic or in the median time from positive blood culture to discharge from the hospital. Thus, requiring preapproval for selected parenteral agents can decrease antimicrobial expenditures and improve susceptibilities to antibiotics without compromising patient outcomes or length of hospital stay. PMID- 9332518 TI - Antibiotic armageddon. PMID- 9332519 TI - Variation of chest radiographic patterns in pulmonary tuberculosis by degree of human immunodeficiency virus-related immunosuppression. The Terry Beirn Community Programs for Clinical Research on AIDS (CPCRA). The AIDS Clinical Trials Group (ACTG). AB - Our aim was to evaluate the effect of human immunodeficiency virus (HIV) disease stage on chest radiographic (CXR) findings among patients with HIV-related pulmonary tuberculosis (TB). Data are from a prospective multicenter treatment trial for HIV-related TB. Baseline CXR findings and CD4+ lymphocyte counts were compared among patients with HIV-related TB. Data from published studies describing CXR findings in HIV-infected patients were reviewed and a pooled-data analysis was conducted. Of 135 patients with culture-confirmed HIV-related TB, 128 had both CXR and CD4+ lymphocyte data. CD4+ lymphocyte counts of < 200/mm3 (n = 98) were significantly associated with hilar/mediastinal adenopathy on CXR (30%, vs. 7% with counts > or = 200/mm3; P = .01); counts of > or = 200/mm3 (n = 30) more frequently were associated with cavitation (20% vs. 7%; P = .08). Analyses of these results, pooled with other published data, confirmed these findings. This study demonstrates associations of certain CXR findings with HIV disease stage. Knowledge of the degree of immunosuppression is important when evaluating CXR findings in HIV-infected patients. PMID- 9332520 TI - Outcomes of bacteremia in patients with cancer and neutropenia: observations from two decades of epidemiological and clinical trials. AB - The prognostic significance of major organ and tissue infection was examined in 909 episodes of bacteremia that were selected from 10 consecutive, randomized clinical trials of antibiotic therapy for infection in patients with cancer and neutropenia. Extensive tissue infection significantly compromised response to initial therapy (38% vs. 74%; P < .0001), ultimate outcome of infection (73% vs. 94%; P < .0001), median time to normalization of temperature (5.3 days vs. 2.5 days; P < .0001), and survival (P < .0001). Other poor prognostic factors revealed by logistic regression included shock (P < .0001) and bacteremia caused by Pseudomonas species (P = .03), Clostridium species (P = .006), or a pathogen resistant to antibiotics used for initial therapy (P < .0001). Recovery of the granulocyte count predicted a superior response (P < .0001). Although the overall mortality rate was not significantly increased when patients with bacteremia due to gram-negative organisms initially received monotherapy or when patients with bacteremia due to gram-positive organisms received delayed vancomycin therapy, these strategies increased the duration of therapy by 25%. Patients with bacteremia due to alpha-hemolytic streptococcus died more often when vancomycin was not included in the initial empirical regimen (P = .004). Because of the prognostic significance of extensive tissue or major organ infection, this factor should be considered in decisions concerning modification of therapy and use of colony-stimulating factors. The cost-effectiveness of initial monotherapy and delayed vancomycin therapy remains to be demonstrated. PMID- 9332521 TI - A clinical and bacteriologic investigation of invasive streptococcal infections in Japan on the basis of serotypes, toxin production, and genomic DNA fingerprints. AB - In a survey of invasive streptococcal infections in Japan, we analyzed isolates of Streptococcus pyogenes collected between 1992 and 1994. Genomic DNA fingerprints produced by pulsed-field gel electrophoresis (PFGE) were compared by computer-assisted analysis. Conventional serologic M types were subdivided into PFGE types showing close genetic similarity. Among the 42 isolates from patients with invasive diseases, 16 PFGE types were identified and genetic diversity was clearly demonstrated. Identical fingerprints were observed in both invasive and noninvasive isolates. Only 43% of invasive isolates produced streptococcal pyrogenic exotoxin A (SPE A), and 31% did not contain the speA gene. These findings suggest that the dissemination of a specific clone is not sufficient to explain all cases of these diseases in Japan and pose a question as to the role of SPE A as a major virulent factor. Bacterial factors other than SPE A and host factors should be considered in evaluation of the pathogenesis of the diseases. PMID- 9332523 TI - Clinical findings for patients with Lyme borreliosis caused by Borrelia burgdorferi sensu lato with genotypic and phenotypic similarities to strain 25015. AB - In the course of performing culture isolation of Borrelia burgdorferi sensu lato for the diagnosis of Lyme borreliosis in Slovenia, we encountered nine patients who were infected with atypical strains. Molecular analyses of these strains suggested that they were more closely related to the North American tick isolate, strain 25015 (which belongs to the DN127 genomic group of B. burgdorferi sensu lato), than they were to the three species (B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii) hitherto found to be associated with European Lyme borreliosis. Review of the case histories of these patients revealed some atypical clinical features and variability in clinical presentation. In this study, we present the clinical findings for these patients and discuss their significance for the diagnosis of Lyme borreliosis. The DN127 genomic group shares with B. burgdorferi sensu stricto the distinction of being present in both the Old and New Worlds. PMID- 9332522 TI - Parasitic sinusitis and otitis in patients infected with human immunodeficiency virus: report of five cases and review. AB - We describe five cases of parasitic sinusitis and otitis in patients infected with human immunodeficiency virus (HIV) and review 14 reported cases. The pathogens identified in our group of patients included agents such as Microsporidium, Cryptosporidium, and Acanthamoeba species. The clinical features common to these patients included a long history of HIV seropositivity associated with advanced immunosuppression and multiple opportunistic infections as well as long-standing local symptoms refractory to multiple courses of antibacterial agents. Symptoms often included fever and chills in addition to local tenderness and discharge. Invasive diagnostic procedures were necessary to obtain the final diagnosis and to initiate appropriate therapy. Although most patients responded at least partially to specific therapy, relapses and recurrences were frequent in patients who did not receive long-term suppressive therapy. The general outcome for HIV-infected patients with parasitic sinusitis and otitis was poor; however, deaths were generally associated with other complications of the underlying HIV infection. PMID- 9332524 TI - Histoplasmosis and kidney disease in patients with AIDS. AB - Renal disease in patients infected with human immunodeficiency virus (HIV) often presents with significant proteinuria and progressive renal failure; focal glomerulosclerosis is the most common renal pathology identified. To our knowledge, we report the first case of nephrotic-range proteinuria and preserved renal function in an HIV-infected patient in association with disseminated histoplasmosis. The initial level of proteinuria was 12.5 g/24 h. The patient developed a concomitant lesion on his neck, which was biopsied and identified as Histoplasma capsulatum by fungal stains and culture. The serum CF titer of antibody against yeast antigens of H. capsulatum was 1:8. The level of serum albumin decreased to 2.0 g/dL, and the level of serum cholesterol increased to 284 mg/dL. Immunohistochemical staining of renal biopsy tissue demonstrated immune complexes within the mesangium; H. capsulatum antigen was also demonstrated in the mesangium. Therapy with oral itraconazole resulted in marked clinical improvement. The findings in this case emphasize the need to rule out treatable causes of the nephrotic syndrome in AIDS, especially in cases of immune complex glomerulonephritis. PMID- 9332525 TI - Retrospective analysis: are fever and altered mental status indications for lumbar puncture in a hospitalized patient who has not undergone neurosurgery? AB - Although nosocomial meningitis is rare in nonsurgical patients, lumbar punctures are frequently performed on hospitalized medical patients who develop delirium and/or fever. A retrospective review was undertaken to determine the yield of lumbar puncture in this setting and to compare it with the yield for suspected community-acquired meningitis. Of 232 lumbar punctures studied, 51 (22%) were performed to rule out nosocomial meningitis, while 181 (78%) were done to rule out community-acquired meningitis. No lumbar puncture performed for suspected nosocomial meningitis was positive, while results of 26 (14%) of those done for suspected community-acquired meningitis were abnormal (P < .01). Patients whose lumbar punctures were positive more often had headache or meningeal signs than those whose lumbar punctures were negative, and only 11 patients (22%) who had lumbar punctures performed for suspected nosocomial meningitis had headache or meningeal signs. We conclude that lumbar punctures performed for suspected nosocomial meningitis in nonsurgical patients have a low yield and that in some low-risk patients without headache or meningeal signs, lumbar puncture may be unnecessary. PMID- 9332526 TI - To tap or not to tap? PMID- 9332527 TI - The clinical diagnosis of genital ulcer disease in men. AB - We report the sensitivity and specificity of physical examination findings for diagnosing primary syphilis, chancroid, and genital herpes. The physical features of genital ulcers in 446 men were measured in accordance with a quantitative scale. Two hundred-twenty of these men had an established, single microbiological diagnosis. Forty-five (20%) had primary syphilis, 118 (54%) had chancroid, and 57 (26%) had genital herpes. There was considerable overlap in the clinical presentation of these three diseases. The classic clinical sign complex attributed to primary syphilis (painless, indurated, clean-based ulcers) was only 31% sensitive but 98% specific. The classic presentation of a chancroid ulcer (a deep, undermined, purulent ulcer) was only 34% sensitive but 94% specific. The classic description of genital herpes ulcers (multiple, shallow, tender ulcers) was only 35% sensitive but 94% specific. Inguinal lymph node findings did not contribute significantly to clinical diagnostic accuracy. These data indicate that the clinical diagnosis of genital ulcer disease can be made with reasonable certainty only for a minority of patients. Rapid, sensitive, and specific diagnostic tests for syphilis, chancroid, and genital herpes are needed. PMID- 9332528 TI - Genital ulceration and clinical acumen. PMID- 9332529 TI - Cutaneous leishmaniasis in the Peruvian Andes: factors associated with variability in clinical symptoms, response to treatment, and parasite isolation rate. AB - The severity of cutaneous leishmaniasis may be determined by host immunity, parasite virulence, and host or vector behavior. We performed a multivariate analysis to identify the main causes of the variability in clinical symptoms, response to treatment, and parasite isolation rate among Peruvian patients. The effect of host immunity was demonstrated first by the finding that secondary infections induced smaller lesions associated with a lower parasite isolation rate than did primary infections and, second, by the finding of fewer lesions in older patients. Phenotypic differences between parasite populations were suggested by the observation that the mean scar size and number varied between villages: patients had more scars in villages where the transmission rates were higher. Human behavior probably determined the site of lesions on the body, since most lesions in the cooler South were on the head, whereas in the North, lesions were equally frequent on the extremities. In addition, older patients, who were more likely infected through occupational exposure, had fewer head lesions. Geographic variation in the pattern of exposure to sandflies indicates that uta control strategies should be region specific. PMID- 9332531 TI - Clinical and epidemiological predictors of recurrent cytomegalovirus disease in orthotopic liver transplant recipients. Boston Center for Liver Transplantation CMVIG Study Group. AB - Predictors of recurrent cytomegalovirus (CMV) disease after the first episode of successfully treated CMV disease in orthotopic liver transplant recipients were studied. Recurrent CMV disease was defined as disease diagnosed > 14 days after the end of a minimum 8-day course of ganciclovir therapy for the first episode and was classified as early or late if it occurred within or after 90 days, respectively, after completion of ganciclovir treatment. Eleven (27%) of 41 patients had recurrent CMV disease (nine early recurrences and two late recurrences). Death was more likely to occur in patients with recurrent CMV disease than in those without it (55% vs. 13%, respectively; P = .006). Initial episodes of multiorgan CMV disease (P = .001) and CMV pneumonia (P = .012) were associated with early recurrence. Multivariate analysis showed that multiorgan CMV disease was independently associated with early recurrence (P = .003; odds ratio, 13.5; 95% confidence interval, 2.4-76.8). Recognition of risk factors for recurrent CMV disease may help identify patients for whom a more intensive therapeutic or diagnostic approach is needed. PMID- 9332530 TI - Disseminated infection due to Bipolaris australiensis in a young immunocompetent man: case report and review. AB - We report a case of disseminated infection due to Bipolaris australiensis in a 21 year-old immunocompetent Pakistani man. He presented with fever and jaundice. Examination revealed a mass in the right lung, mediastinal lymphadenopathy, a pericardial effusion, and abdominal masses obstructing and invading the common bile duct and right ureter. Histological examination and culture of a biopsy specimen of the hilar mass yielded the fungal pathogen B. australiensis. The patient was treated successfully with amphotericin B and itraconazole. PMID- 9332532 TI - Incidence and epidemiology of nosocomial infections in patients infected with human immunodeficiency virus. AB - In a prospective study, we investigated the incidence, characteristics, and risk factors of nosocomial infections (NIs) in patients with human immunodeficiency virus disease. There was a total of 528 admissions of 405 eligible patients; 46 NIs (8.7% per discharge) were identified in 39 patients. The proportional frequencies of NIs were as follows: 16 skin and/or soft-tissue infections (including localized catheter-associated infections), 3.0%; 14 respiratory tract infections, 2.7%; 11 bloodstream infections, 2.1%; and 5 urinary tract infections, 0.9%. The most common etiologic agents were Staphylococcus aureus (27.6%), Pseudomonas aeruginosa (13.8%), and Enterobacter cloacae (13.8%). The duration of hospitalization was not significantly prolonged by NI in the cohort. PMID- 9332533 TI - Changing epidemiology of nosocomial infections in human immunodeficiency virus infected patients. PMID- 9332534 TI - Disseminated Flavimonas oryzihabitans infection in a diabetic patient who presented with suspected multiple splenic abscesses. PMID- 9332535 TI - Pseudomonas stutzeri community-acquired pneumonia associated with empyema: case report and review. PMID- 9332536 TI - Necrotizing fasciitis caused by unencapsulated Haemophilus influenzae. PMID- 9332537 TI - Levels of transforming growth factor beta 1, tumor necrosis factor alpha, and interleukin 6 in cerebrospinal fluid: association with clinical outcome for children with bacterial meningitis. PMID- 9332538 TI - Increased endothelin levels in cerebrospinal fluid samples from adults with bacterial meningitis. PMID- 9332539 TI - Apophysomyces elegans limb infection with a favorable outcome: case report and review. PMID- 9332540 TI - Measurement of human immunodeficiency virus (HIV) type 1 RNA load distinguishes progressive infection from nonprogressive HIV-1 infection in men and women. PMID- 9332541 TI - Chromobacterium violaceum infection of the deep neck tissues in a traveler to Thailand. PMID- 9332542 TI - Clinical and pathophysiological aspects of immune complex glomerulonephritis associated with Entamoeba histolytica abscess of the liver. PMID- 9332543 TI - Spontaneous rupture of the spleen revealing primary human immunodeficiency virus infection. PMID- 9332544 TI - Recurrent iritis after intravenous administration of cidofovir. PMID- 9332545 TI - Rapid emergence of resistance to cefepime during treatment. PMID- 9332546 TI - Relapsing infection due to Enterobacter species: lessons of heterogeneity. PMID- 9332547 TI - Disseminated histoplasmosis and human immunodeficiency virus type 1 infection: risk factors in Guatemala. PMID- 9332548 TI - Enterocytozoon bieneusi infection in patients who are not infected with human immunodeficiency virus. PMID- 9332549 TI - Effect of food intake on the bioavailability of itraconazole. PMID- 9332550 TI - Inadequate use of immunization to prevent severe pneumococcal infection. PMID- 9332552 TI - Changing the focus of infectious diseases to pathogen-specific therapy. PMID- 9332551 TI - Fluoroquinolone prophylaxis for the prevention of bacterial infections in patients with cancer--is it justified? PMID- 9332553 TI - Apolipoprotein E4 and Alzheimer's disease in Sao Paulo-Brazil. AB - Several recently published studies showed the existence of an association between the allele epsilon 4 of the apolipoprotein E and Alzheimer's disease (AD) in developed countries. We examined this association in 55 patients with possible or probable AD and 56 elderly controls referred to outpatient clinics at the "Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo" and "Centro de Saude Escola da Faculdade de Saude Publica da Universidade de Sao Paulo". The allele epsilon 4 was significantly more frequent among patients than controls (20.9% vs 8.9%, p = 0.038). Thirty-six percent of the cases presented with at least one allele epsilon 4 compared with only 17.9% of the controls (p = 0.027). The presence of at least one epsilon 4 allele increased by 2.63 times the risk of subjects being diagnosed as suffering from AD. All three epsilon 4 epsilon 4 patients were male and had a pre-senile onset of the disease. There was no significant difference between senile and pre-senile cases (41.9% vs 29.2%, p = 0.326) nor between men and women (36.0% vs 36.7%, p = 0.959) regarding their risk of being epsilon 4. The age at onset of symptoms did not differ among the different genotype groups, although epsilon 4 epsilon 4 cases showed a consistent trend for earlier onset. When only patients with the diagnosis of "probable AD" were included in the analysis (n = 43), we observed that 22.1% of the alleles were epsilon 4, a rate that was significantly higher than the 8.9% of controls (p = 0.024). This study supports the association between the presence of the epsilon 4 allele and AD and extend this finding to Brazilian patients. Nonetheless, the presence of this allele is not necessary nor sufficient for the development of the disease and it is possible that its contribution to the pathogenesis of the disorder depends on the subject's ethnic group. PMID- 9332554 TI - Human neurocysticercosis. IgE in cerebrospinal fluid. AB - The detection of IgE is technically difficult because of its reduced concentrations in serum, and even lower concentrations in cerebrospinal fluid (CSF). In the present investigation we studied 86 CSF samples using an immunoenzymatic method with an anti-IgE-alkaline phosphatase conjugate and a fluorigenic substrate. The samples were from three groups: A) 29 patients with neurocysticercosis (NC), B) 36 patients with different neurologic disorders (neurosyphilis, neurotuberculosis, meningitis, tumors, hemorrhage) and C) 21 discharged individuals who had been hospitalized for bacterial meningitis. The results obtained were: A) 0.05 to 3.00 IU/ml (0.76 +/- 0.79), B) 0.00 to 1.50 IU/ml (0.23 +/- 0.34) and C) 0.05 to 1.25 IU/ml (0.34 +/- 0.34). The present results suggest that IgE appears to play a role in the pathogeny of NC and that efforts should be made to standardize a test for the detection of specific IgE antibodies. PMID- 9332555 TI - Mumps meningoencephalitis. An epidemiological approach. AB - The aim of this study was to analyse distribution of meningoencephalitis caused by mumps virus in children related to sex, age and seasonal influences. Thirty seven children were evaluated, ages ranging from 2 to 14 years. They were seen at Emergency Unit of Faculdade de Medicina do Triangulo Mineiro and at Hospital da Crianca, in Uberaba-MG, Brazil, from March 1st 1991 to February 1st 1993 and they were hospitalized for about 5 days. Through a protocol findings were studied during hospitalization and clinical course stressing epidemiology, symptomatology, cerebrospinal fluid studies, electroencephalogram and cortical function analysis. Only epidemiological data were considered in the present study. Data analysis revealed male predominance, at a range from 5 to 9 years and great number of occurrences at the last quarter of the year. PMID- 9332556 TI - Middle cerebral artery revascularization. Anatomical studies and considerations on the anastomosis site. AB - In the surgical management of skull base lesions and vascular diseases such as giant aneurysms, involvement of the internal carotid artery may require the resection or the occlusion of the vessel. The anastomosis of the external carotid artery and the middle cerebral artery with venous graft may be indicated to re establish the blood flow. To determine the best suture site in the middle cerebral artery, an anatomical study was carried out. Fourteen cerebral hemispheres were analysed after the injection of red latex into the internal carotid artery. The superior and inferior trunk of the main division of the middle cerebral artery have more than 2 mm of diameter. They are superficial allowing an anastomosis using a venous graft. The superior trunk has a disadvantage, it gives rise to branches for the precentral and post-central giri. The anastomosis with the inferior trunk presents lower risk of neurological deficit even though the angular artery originates from it. PMID- 9332557 TI - [Cerebral extraction of oxygen. A practical model and its clinical applications]. AB - Cerebral extraction of oxygen (CEO2) represents a practical physiologic measure, with multiple clinical applications. This variable is calculated as the arterio jugular difference in oxyhemoglobin saturation (from arterial blood and from the jugular bulb). Because it involves global measurements, the CEO2 does not allow detection of regional abnormalities in cerebral hemometabolism. However, when dealing with predominantly (not exclusively) global changes, the CEO2 provides accurate information on the balance (or coupling) between cerebral consumption of oxygen and cerebral blood flow. PMID- 9332558 TI - [Measurement of arteriovenous oxygen difference in the monitoring of patients with subarachnoid hemorrhage due to cerebral aneurysm]. AB - The arterious venous oxygen difference (AVDO2) due to the close relationship with cerebral metabolic rate of oxygen and cerebral blood flow shows metabolic alterations that occur in some pathological situations in the brain including subarachnoid haemorrhage. The AVDO2 was calculated by the Fick equation and the results evaluated by the Glasgow outcome scale. Measurements of arteriojugular oxygen difference were carried out in 30 patients with subarachnoid haemorrhage due to rupture of intracranial aneurysms, as an attempt to monitor the relationship between changes in AVDO2, clinical picture, and evolution of the patients. The subarachnoid haemorrhage was diagnosed by CT scan in 17 patients and by lumbar punction in 13 and the diagnosis of arterial vasospasm was carried out by clinical evaluation and confirmed by four vessels angiogram in only eight patients. Eighteen patients were admitted with Hunt & Hess (H&H) I/II, seven with H&H III and five with H&H IV/V. Nineteen patients had AVDO2 normal and this group had three deaths; five patients had AVDO2 continuously low with three deaths; and six patients had AVDO2 continuously high with two deaths. The patients with normal AVDO2 had better prognosis and clinical evolution than the patients with abnormal values of AVDO2. In conclusion, AVDO2 measurements could not be correlated with the diagnosis of vasospasm, but was useful in the early identification of metabolic changes that occur after subarachnoid haemorrhage and could be used as an supplementary monitoring in the clinical evaluation of patients with this pathology. PMID- 9332559 TI - [Subsidiary predictive tests in epileptic crisis after ischemic stroke]. AB - We studied subsidiary laboratorial tests such as serum glucose, red blood cell count, total cholesterol, HDL and LDL cholesterol and triglycerides, electrocardiogram, electroencephalogram (EEG), cerebrospinal fluid, and CT scan of 35 patients with cerebral infarction who developed epileptic seizures (group 1 or G1), and compared them to a group of 35 patients who had cerebral infarction but have not developed epileptic seizures (group 2 or G2). The EEG analysis showed significance in the comparison between the groups; focal identification of the electrical cerebral activity was the most frequent abnormality found in G1. Extensive infarcts were also more frequent in G1. The association of abnormal EEG and extensive lesion on CT may be considered a preditive factor for occurrence of epileptic seizures after cerebral infarction. The analysis of the other tests showed no significance on the comparison between the groups. PMID- 9332561 TI - [Normative data on the verbal fluency test in the animal category in our milieu]. AB - OBJECTIVE: Evaluate the performance on verbal fluency (VF) in our population in a Brazilian sample checking the influence of age and literacy. METHODS: 336 people without neurological or psychiatric complaints evaluated through Mini-Mental State Examination and VF (animals). For comparison, and to determine cut-off points, 65 people with cognitive loss followed at our clinic were also evaluated. RESULTS: We found a mean of 13.8 animals in 1 minute, with the following distribution: illiterates, 11.9; up 4 years of education, 12.8; 4 to 7 years, 13.4; 8 years or more, 15.8 (p = 0.0001). In relation to age the means were: up to 64 years, 13.7; 65 years or more, 13.9. There was no difference between the two groups. The cut-off points were 9 for people under 8 years of education with a sensitivity of 75% for illiterates, 100% for low educational level (up 4 years), and 87% for middle level (4 to 7 years). The specificity was respectively 79%, 84%, and 88%. For the high educational level the mean was 13 with a sensitivity of 86% and specificity of 67%. CONCLUSIONS: In the VF (animals) there is a significant influence of schooling and different cut-off points should be used. PMID- 9332560 TI - [Clinical and laboratory changes before the development of delirium tremens]. AB - Thirteen alcoholic male patients that developed delirium tremens (DT) after admission in a psychiatric hospital for treatment of alcoholism (group I) had their clinical and laboratorial records examined. The laboratory samples were taken during the phase previous at the DT. Data on this group were compared to those of two other groups of alcoholics--26 patients each--that did not develop DT in the present admission, with (group II) or without (group III) previous history of DT. The patients of group I had significantly lower average age and worse general conditions than the patients of group III. The frequency of elevated aminotransferases and hypomagnesemia was significantly higher in the group I and II than in the group III. The aminotransferases, especially the aspartate-aminotransferase, were significantly more elevated in the groups I and II. PMID- 9332562 TI - [Normalization of a computerized visual attention test (TAVIS)]. AB - The authors present the normative data of a computerized test (TAVIS) that address visual attention in children and adolescents being the first neuropsychological instrument as such devised and developed in Brazil. Selective, alternate and sustained attention aspects are evaluated through three different tasks. Omission and action errors as well as time reaction are evaluated. The advantages and limitations of the test are commented. PMID- 9332563 TI - [Magnus-De Kleijn tonic neck reflex]. AB - The Magnus-De Kleijn's tonic neck reflex is analyzed concerning to the developmental psychologies of Gesell, Spitz and Piaget. It is considered its phylogenetic nature, it is taken into account its favorable disappearing about three months old and, it is made a great account of its participation in baby development. PMID- 9332564 TI - [Infections of cerebrospinal fluid shunts in children. Review of 100 infections in 87 children]. AB - An analysis of 100 infections in 87 children treated with shunts in the period of 1982 to 1995 is reported. The clinical presentation has been more frequently secondary to inflammatory signals. Staphylococcus were the most frequently microorganisms found. Infection by Gram negative agents was more aggressive and directly related with failure of therapy. Treatment included since only systemic antibiotics until withdrawal of shunt with use of systemic and intrathecal antibiotics. The best therapeutic results were obtained with withdrawal of shunt system and replacement by external shunt system associated to systemic antibiotics. In our experience this management must be accepted for treatment of this severe complication. PMID- 9332566 TI - [Acromegaly. Diagnostic and therapeutic aspects. Analysis of 18 cases]. AB - The authors present a retrospective study of a series of eighteen patients with acromegaly diagnosed, treated and followed by the Endocrinology and Neurosurgery Services of the Hospital Universitario Clementino Fraga Filho of the Federal University of Rio de Janeiro. The average age of the patients was 43.2% years (varying between 15 and 63). Initial complaints were mainly due to somatic alterations in 83.33%; half the cases had manifestations secondary to tumor compression and 53.33% had neuro-ophtalmological alterations. Hypersecretion of growth hormone was demonstrated by basal hormone determinations and dynamic tests. Neuroradiological assessment showed supraselar expansion in 61.11% of cases. Surgical approach was transsphenoidal in all cases. The main objective of this study was to establish diagnostic criteria, discuss the therapeutic conduct and evaluate the results obtained, comparing them with other series of literature. PMID- 9332565 TI - [Cerebellar astrocytomas in childhood. Experience with 25 cases]. AB - The experience with the surgical treatment of cerebellar astrocytomas in 25 children is reported. The clinical presentation, incidence, CT-scan diagnostic studies, pathology, recurrence and treatment aspects are discussed. The series included children until 10 years old with peak (7 cases) in the 7th year of age. The more frequent opening symptoms were: headache, vomit and gait disturbances. No surgical mortality occurred in the series. The authors conclude that surgical radical resection is the best therapeutics for this type of tumor and that radiotherapy is indicated only for tumors with malignant histology. PMID- 9332567 TI - [Characteristics of the patients with head injury assisted at the Hospital das Clinicas of the Ribeirao Preto Medical School]. AB - The chart of 3468 patients with head injury assisted in the Hospital das Clinicas Ribeirao Preto Medical School, from 1990 through 1992 were analyzed aiming to determine their main characteristics. Regarding sex, there was predominance of male. Accidental fall among children and traffic accidents among adults were the main causes of trauma. Daily distribution of assistance revealed an increase between 8 and 12 PM and during the week there was a constant flow from Tuesday to Friday and progressively increased on Saturday to Sunday. Approximately 75% of the patients presented mild head injury (score equal or superior to 13 in the Glasgow Coma Scale). Headache among children and vomiting, headache and alcoholic abuse among adults were the most frequent signs and symptoms at admission. At discharge 87.2% of patients had no symptoms and mortality was 5.7%. Peculiarities of head injury in Ribeirao Preto are discussed. PMID- 9332569 TI - Effect of maternal protein deprivation on morphological and quantitative aspects of the myenteric plexus neurons of proximal colon in rats. AB - We have studied the morphological and quantitative aspects of the myenteric plexus neurons of the proximal colon in rats (Rattus norvegicus of Wistar strain) submitted to a protein deprivation during prenatal and lactation periods. Twenty pregnant dams were divided in four groups labeled according to the kind of nourishment they were given: Group NN, normal diet; Group DN, low protein diet during prenatal period, and normal diet during lactation period; Group ND, normal diet during prenatal period, and low protein diet during lactation period; Group DD, low protein diet during prenatal and lactation periods. Histological analyses were developed with proximal colon segments using the haematoxylin and eosin staining method. Membrane preparations were stained by Giemsa's method. The statistical analysis has demonstrated no significant difference among the means of neurons found in the four studied groups. It was noticed that the animals under protein deprivation during prenatal and lactation periods presented greater quantity of large and strongly basophilic myenteric neurons. This suggests that neurons have accumulated protein in the cytoplasm. PMID- 9332568 TI - [Tolosa-Hunt syndrome. Troubles in diagnosis and pattern of response to prednisone]. AB - The Tolosa-Hunt syndrome (THS) consists of a painful ophthalmoplegia related to granulomatous inflammatory process in the cavernous sinus. According to recent concepts, the diagnosis is established only when other causes of painful ophthalmoplegia are ruled out. A typical pattern of response to corticosteroids associated with a benign evolution may reinforce this possibility. Tumors such as lymphoma and meningioma and orbital pseudotumors can make difficult the differential diagnosis because they also may respond to steroids. Thus it is always necessary to make an extensive ancillary investigation. We performed a clinical, laboratory and radiologic study of five patients with THS. Prednisone was used in all, with dosages ranging from 40 to 80 mg/day. In four patients there was a dramatic analgesic effect in less than 48 hours. Improvement of the ophthalmoplegia was not so fast but occurred in all with a complete remission in 4 to 45 days. PMID- 9332570 TI - [Anatomo-pathological and ultrastructural features of mucopolysaccharidosis. Case report]. AB - The mucopolysaccharidoses (MPS) are lysosomal storage diseases in which a specific enzyme defect causes glycosaminoglycans storage in tissues. The authors present a necropsy case of a 10 years old boy with clinical and laboratorial diagnosis of MPS. The necropsy revealed thickening of meninges, cardiac valves and hepatomegaly. The microscopical examination of the brain showed finely vacuolated histiocytes around blood vessels and meninges. Systemic deposits of vacuolated histiocytes in cardiac valves and liver were also detected. The ultrastructural examination of the brain, liver and spleen showed filamentous material accumulated in vacuolated histiocytes and hepatocytes and features neuronal storage disease. PMID- 9332571 TI - Progressive multifocal leukoencephalopathy in a child with acquired immunodeficiency syndrome (AIDS). AB - Progressive multifocal leukoencephalopathy is a rare viral-induced demyelinating disease associated to immunodeficiency. A 10-year-old boy with AIDS is reported, who developed subacute cerebellar signs and symptoms with multiple cranial nerve involvement and dementia. A computed tomography scan revealed a focal nonenhancing area of low attenuation in the cerebellum. On magnetic resonance imaging high signal lesions in T2 weighted sequences were shown. The biopsy of one of those lesions showed the typical histological findings of progressive multifocal leukoencephalopathy. It seems important to consider this diagnosis in children with AIDS who present with progressive neurological features. PMID- 9332572 TI - [Neurotoxoplasmosis presenting as multifocal suppuration and hemorrhage. Case report]. AB - Neurotoxoplasmosis is one of the commonest infections in immunosuppressed patients, and rarely presented in the hemorrhagic form. We describe a case with multiple nodular hemorrhagic lesions with perilesional edema in the computed tomography. The neuropathological study showed focal lesions, necrosis and microorganisms compatible with toxoplasmosis. PMID- 9332573 TI - [Possible role of the immune system in lead peripheral polyneuropathy. Case report]. AB - This is a case report of a twenty-five years old man who developed, due to lead intoxication, a severe axonal peripheral predominantly motor neuropathy, after a shotgun injury. The projectile was retained in the right hip. Before this diagnosis had been done he was treated with corticosteroids in immunosuppressive doses and showed an improvement, but he had worsened at each attempt to interrupt the drug. Because he had also other signs of lead intoxication, such as abdominal cramps, severe anemia and seizures it was search for the blood levels of lead that was 101.2 micrograms/dl. The patient was treated with calcium disodium edetate and surgical removal of lead fragments. After that he had a good outcome with no need of corticosteroids. It is emphasized the possible relevance of the immune system on the mechanism of plumbic intoxication and the importance of the withdrawal of the lead material retained in joints. PMID- 9332574 TI - Acute orbital myositis. Case report. AB - The case of 22-year old, white woman with bilateral orbital myositis following an acute upper respiratory tract infection is reported. The most important clinical findings were ocular pain, proptosis, restricted eye motility and swelling of the eyelids. The enlarged eye muscles were seen on orbital computerized tomography scan. The clinical findings of inflammatory orbital myositis and clinical response to corticotherapy are emphasized. PMID- 9332575 TI - [Clinical and neuroradiologic aspects of pseudohypoparathyroidism. Case report]. AB - The authors describe the case of a 18-year-old man with short stature, epilepsy, mental deficiency and basal ganglia and central nervous system calcifications. The clinical and laboratorial findings have suggested pseudohypoparathyroidism which is a rare pathology with a peripheral resistance to parathormone, neuromuscular hyperexcitability, short stature and various clinical findings. This paper reviews the clinical form and treatment of pseudohypoparathyroidism and the neuroradiologic aspects of calcifications. PMID- 9332576 TI - [Cerebrospinal fluid analysis and the pathogenesis of central nervous system infection by HTLV-I]. AB - The immunopathogenesis of the HTLV-I associated myelopathy (HAM) may be studied by the CSF evaluation. The mechanism of this myelopathy remains unknown. The disturbs of the cellular and humoral immune response observed in HAM patients suggest that the immunological derangement may contribute to the disease mechanisms. For hypothesis, the migration of infected lymphocytes through the blood-brain barrier could have a main role at the pathogenesis of HAM. An increase of the production of cytokines as tumor necrosis factor alpha (TNF alpha) contributes to the migration of lymphocytes through the expression of the intercellular adhesion molecule on the surface of the endothelial cells. On the other side, new knowledges suggest that the imbalance between the production of TNF alpha and its soluble receptor (sTNF-R) could result in the lesive effects of this cytokine in the central nervous system. PMID- 9332577 TI - [Peripheral neuropathies in the State of Sao Paulo from 1939 to 1985]. AB - Studies on peripheral neuropathies by investigators residing in the State of Sao Paulo, Brazil, and published since the 1930 and 1940 decades until 1985 were revised in the present survey. Investigations in the area were greatly encouraged by the appearance of the journal Arquivos de Neuro-Psiquiatria(Sao Paulo). Oswaldo Freitas Juliao may be considered the author who began these studies in the State and his most important contributions were related to leprosy and to Andrade disease, although he also published papers on other types of peripheral neuropathies. Horacio Martins Canelas also made a very important contribution to the study of different neuropathies, especially those due to vitamin B12 deficiency. A series of papers on neuropathies published by neurologists residing in the State is summarized. We also present a catalogue of the major university centers where groups of neurologists preferentially devote their time to the study of neuromuscular disease in Sao Paulo and a selected bibliography about neuropathies by investigators from this State. PMID- 9332578 TI - Re-evaluation of schistosomiasis mansoni in Minas Gerais, Brazil--II. Alto Paranaiba mesoregion. PMID- 9332579 TI - Age structure of adult mosquito (Diptera: Culicidae) populations from Buenos Aires Province, Argentina. AB - In order to detect seasonal trends in the age structure of adult mosquitoes from Buenos Aires province, Argentina, female populations were sampled with CDC traps during 1989-1991 in Punta Lara and La Plata. The mosquitoes were dissected and age-grouped according to ovarian tracheation and ovariolar stages. All Runchomyia paranensis females were parous, suggesting that this species could be autogenous. Aedes albifasciatus showed parous peaks following population peaks, with shorter delays in spring-summer and longer in fall-winter. Ae. crinifer and Culex dolosus showed wide fluctuations in age structure due to adult emergences during all months. Psorophora ferox showed high population replacement rates. Mansonia indubitans and Ma. titillans have few generations per year during their activity period. This is the first report on age-grading of adults of field mosquito populations from Argentina. PMID- 9332580 TI - Infection of Triatoma guasayana, Triatoma sordida and Triatoma infestans by Trypanosoma cruzi from a naturally infected opossum. PMID- 9332581 TI - The potential for dispersal of onchocerciasis in Ecuador in relation to the distribution of the vector Simulium exiguum (Diptera:Simuliidae). AB - The future dispersal of onchocerciasis in Ecuador is dependent on the distribution of cytotypes of the vector species complex Simulium exiguum. Over the last 14 years, collections of larvae have been made from over 25 rivers, between 80-1600 m altitude, from various sites on both sides of the Andes. Analysis of larval polytene chromosomes was used to determine the distributions of each cytotype. On the western side of the Andes, the Cayapa cytotype (the only cytotype directly incriminated as a vector) has a distribution from Santo Domingo de los Colorados northwards. The Quevedo and Bucay cytotypes occur from Santo Domingo de los Colorados southwards. On the eastern side of the Andes, the Aguarico cytotype occurs in the Rio Aguarico and a new cytotype is present in the tributaries of the Rio Napo. Whether the disease will spread south of Santo Domingo and on the eastern side of the Andes depends on vector capacity of the cytotypes and the dispersal patterns of individuals infected with onchocerciasis. At present the Aguarico, Bucay and Quevedo cytotypes are known to be efficient hosts, but their biting preferences and biting densities have not yet been evaluated. PMID- 9332583 TI - Wild reservoirs infected by Trypanosoma cruzi in the ecological park "El Zapotal", Tuxtla Gutierrez, Chiapas, Mexico. PMID- 9332582 TI - Onchocerciasis in Ecuador: prevalence of infection on the Ecuador-Colombia border in the Province of Esmeraldas. AB - The prevalence of onchocerciasis infection was determined in communities on 7 rivers located in the northern area of the canton San Lorenzo, province of Esmeraldas. Diagnosis of the infection was obtained by skin biopsies and recombinant-antigen based-serology. No evidence of infection was detected in 9 communities studied along the Rio Mataje, which forms the frontier between Ecuador and Colombia, nor in 10 adjacent communities located on 5 interior rivers. Evidence for Onchocerca volvulus infection was found in 4 communities on the Rio Tululvi with the following prevalence: La Boca (3.5% by biopsy and 3.9% by serology), Guayabal (9.1% by both biopsy and serology), La Ceiva (51.5% by biopsy and 53% by serology), and Salidero (4% by biopsy and 7.7% by serology). A few individuals in these communities were seropositive for O. volvulus in the absence of detectable dermal microfilariae: these might harbor very light or prepatent infections. No clinical disease attributable to onchocerciasis was found. The infected communities will be included in the ivermectin-based National Control Program for the disease, with no evidence of the infection having extended north of the Ecuadorian-colombian border. PMID- 9332584 TI - Antibodies anti bloodstream and circumsporozoite antigens (Plasmodium vivax and Plasmodium malariae/P. brasilianum) in areas of very low malaria endemicity in Brazil. AB - During 1992-1994, 33 malaria cases were reported in two regions in Brazil where few sporadic atypical cases occur, most of them in home owners, who are weekenders, while home caretakers live there permanently. Indirect Fluorescent Antibody Test (IFAT), with Plasmodium vivax, and Enzime Linked Immunosorbent Assay (ELISA) with repeat peptides of the circumsporozoite (CS) proteins of the 3 known P. vivax variants and P. malarie/P. brasilianum, were performed on 277 sera, obtained within a 5 to 10 km range of malaria cases. Very rarely did any of these donors recall typical malaria episodes. Blood smears of all but 5 were negative. One of the 5 malaria cases included in our serology was of a home owner, I of a permanent resident, 3 from Superintendencia de Controle de Endemias employees who went there to capture mosquitoes. In Region 1 the prevalence of IFAT positive sera was 73% and 28% among caretakers, 18% and 9.6% among home owners. In Region 2 (3 localities) no distinction was possible between caretakers and home owners, IFAT positivity being 38%, 28% and 7%. The relative percentage of positive anti-CS repeats ELISA, differed for each of the peptides among localities. Dwellings are in the vicinity of woods, where monkeys are frequently seen. The origin of these malaria cases, geographical differences and high seropositivity is discussed. PMID- 9332585 TI - Tuberculin skin test and CD4+/CD8+ T cell counts in adults infected with the human immunodeficiency virus in Medellin, Colombia. AB - To evaluate the effect of BCG vaccination and T lymphocyte subpopulations on the reactivity to the tuberculin skin test, 113 asymptomatic HIV+ individuals were tuberculin tested by intradermal injection of 5TU of purified protein derivative and the levels of circulating lymphocyte (CD3, CD4 and CD8) subpopulations determined by indirect immunofluorescence. Ninety-two percent of the subjects included in the study were males. The mean age of the group was 32.1 +/- 7.4 years. Sixty-two percent presented a BCG scar. However, only 22% exhibited positive tuberculin reactions (> or = 5 mm) irrespective of the presence of the BCG scar. Tuberculin positive individuals exhibited higher CD4+ cell counts (p = 0.004) and CD4+/CD8+ ratios (p = 0.006) than tuberculin negative (< 5 mm) HIV+ individuals. The number of individuals with positive tuberculin reactions was significantly higher in subjects with more than 500 CD4+ lymphocytes/microliter (p = 0.02) or CD4+/CD8+ ratios > or = 1.12 (p = 0.002). These results suggest that a prior BCG vaccination does not influence the reactivity to the tuberculin skin test in HIV+ asymptomatic individuals and that the number of CD4+ lymphocytes and the CD4+/ CD8+ ratio positively correlate with the tuberculin reactivity. PMID- 9332586 TI - In vitro responses of Plasmodium falciparum isolates to five antimalaria drugs in French Guiana during 1994 and 1995. PMID- 9332587 TI - Simplified method for preservation and polymerase chain reaction-amplification of Trypanosoma cruzi DNA in human blood. PMID- 9332588 TI - Cloning, expression and toxicity of a mosquitocidal toxin gene of Bacillus thuringiensis subsp. medellin. AB - Bacillus thuringiensis (Bt) subsp. medellin (Btmed) produces parasporal crystalline inclusions which are toxic to mosquito larvae. It has been shown that the inclusions of this bacterium contain mainly proteins of 94, 68 and 28-30 kDa. EcoRI partially digested total DNA of Btmed was cloned by using the Lambda Zap II cloning kit. Recombinant plaques were screened with a mouse polyclonal antibody raised against the 94 kDa crystal protein of Btmed. One of the positive plaques was selected, and by in vivo excision, a recombinant pBluescript SK(-) was obtained. The gene encoding the 94 kDa toxin of Btmed DNA was cloned in a 4.4 kb DNA fragment. Btmed DNA was then subcloned as a EcoRI/EcoRI fragment into the shuttle vector pBU4 producing the recombinant plasmid pBTM3 and used to transform by electroporation Bt subsp. israelensis (Bti) crystal negative strain 4Q2-81. Toxicity to mosquito larvae was estimated by using first instar laboratory reared Aedes aegypti, and Culex quinquefasciatus larvae challenged with whole crystals. Toxicity results indicate that the purified inclusions from the recombinant Bti strain were toxic to all mosquito species tested, although the toxicity was not as high as the one produced by the crystal of the Btmed wild type strain. Polyacrylamide gel electrophoresis indicate that the inclusions produced by the recombinant strain Bti (pBTM3) were mainly composed of the 94 kDa protein of Btmed, as it was determined by Western blot. PMID- 9332589 TI - Biomorphological alterations induced by an anti-juvenile hormonal compound, 2-(2 ethoxyethoxy)ethyl furfuryl ether, on three species of triatominae larvae (Hemiptera, Reduviidae). AB - Applied topically to larvae of Rhodnius prolixus Stal, Triatoma infestans (Klug) and Panstrongylus herreri Wygodzinsky, vectors of Trypanosoma cruzi, the causative agent of Chagas' disease, a synthetic, furan-containing anti-juvenile hormonal compound, 2-(2-ethoxyethoxy)ethyl furfuryl ether induced a variety of biomorphological alterations, including precocious metamorphosis into small adultoids with adult abdominal cuticle, ocelli, as well as rudimentary adultoid wings. Some adultoids died during ecdysis and were confined within the old cuticle. The extension of these biomorphological responses is discussed in terms of the complexity of the action of anti-juvenile hormonal compounds during the development of triatomines. PMID- 9332590 TI - A preliminary analysis of insects of medico-legal importance in Curitiba, State of Parana. AB - A survey of the carrion fauna was made at two sites in Curitiba, State of Parana, with the objective of describing the insects associated with carrion and setting up a preliminary data-base for medico-legal purposes in south Brazil. Vertebrate exclusion experiments were carried out in each season between 1994 and 1995 with a 250 g laboratory-bred rat (Rattus norvegicus). Five stages of decomposition were identified: fresh, bloated, decaying, dry and adipocere-like. Some species showed seasonal and site preference and so could be used to identify the probable place and season where death took place. Sarconesia chlorogaster (Diptera, Calliphoridae) was restricted to an open field site and to cooler months. Hemilucilia semidiaphana (Diptera, Calliphoridae) and Pattonella resona (Diptera, Sarcophagidae) were restricted to the forest site and warmer months. Phaenicia eximia (Diptera, Calliphoridae) and Oxyletrum discicolle (Coleoptera, Silphidae) were present at both sites throughout the year and could be useful for population level analysis. Dissochaetus murray (Coleoptera, Cholevidae) was present throughout the year at the forest site and was associated with the adipocere-like stage. Ants played an important role producing post-mortem injuries to the carcasses. Insects of 32 species are reported as being useful in community level approaches. PMID- 9332591 TI - [Various aspects of the biology of Triatoma pseudomaculata Correa & Espinola, 1964, in laboratory conditions (Hemiptera:Reduviidae:Triatominae)]. AB - Observations were made on the evolutive cycle of Triatoma pseudomaculata, held under laboratory conditions, fed weekly on bird (pigeon). Of 60 eggs obtained, only 34 nymphs reached the adult stage in a period of X(S) = 398 +/- 76 days. The following parameters were observed: the time immature stages took to develop from egg to adult emergence; the occurrence of the first meal; the time-lapse between the presenting of the blood-meal and the beginning of feeding; time of feeding; amount of blood ingested; variation of weight 24 hr after the blood-meal and until the next blood-meal; and the defecation pattern. The experiment was carried out for 20 months, held in BOD incubator with the average of temperature and humidity of 28 +/- 1 degrees C and 80 +/- 5% RU, respectively. PMID- 9332592 TI - The changing face of the epidemiology of tuberculosis due to molecular strain typing--a review. AB - About one third of the world population is infected with tubercle bacilli, causing eight million new cases of tuberculosis (TB) and three million deaths each year. After years of lack of interest in the disease, World Health Organization recently declared TB a global emergency and it is clear that there is need for more efficient national TB programs and newly defined research priorities. A more complete epidemiology of tuberculosis will lead to a better identification of index cases and to a more efficient treatment of the disease. Recently, new molecular tools became available for the identification of strains of Mycobacterium tuberculosis (M. tuberculosis), allowing a better recognition of transmission routes of defined strains. Both a standardized restriction-fragment length-polymorphism-based methodology for epidemiological studies on a large scale and deoxyribonucleic acids (DNA) amplification-based methods that allow rapid detection of outbreaks with multidrug-resistant (MDR) strains, often characterized by high mortality rates, have been developed. This review comments on the existing methods of DNA-based recognition of M. tuberculosis strains and their peculiarities. It also summarizes literature data on the application of molecular fingerprinting for detection of outbreaks of M. tuberculosis, for identification of index cases, for study of interaction between TB and infection with the human immuno-deficiency virus, for analysis of the behavior of MDR strains, for a better understanding of risk factors for transmission of TB within communities and for population-based studies of TB transmission within and between countries. PMID- 9332593 TI - Chagas disease in Ecuador: evidence for disease transmission in an Indigenous population in the Amazon region. AB - Two well-defined synthetic peptides TcD and PEP2 were used in a sero epidemiological study for the detection of Trypanosoma cruzi infections in an indigenous group in the Amazon region of Ecuador. Of the 18 communities studied along the Rio Napo, province of Napo, 15 (83.3%) were found to be positive for T. cruzi infection. Of the 1,011 individuals examined 61 (6.03%) resulted positive. A prevalence of infection of 4.8% was found in children aged 1-5 years. The prevalence of infection increased with age, with adults 50 years or older showing a maximum prevalence of 18.8%. Autochthonous transmission of T. cruzi is present among this isolated indigenous population. PMID- 9332594 TI - Blastocystis hominis infection in Cuban AIDS patients. PMID- 9332595 TI - Evaluation of chemical and physical-morphological factors as potential determinants of Biomphalaria pfeifferi (Krauss, 1848) distribution. AB - This study was carried out in five sites along a small perennial river system in south-central Tanzania, which had been identified as the focus for transmission of intestinal schistosomiasis in the area. Malacological surveys preceding the study showed a focal distribution of Biomphalaria pfeifferi, intermediate host snail of Schistosoma mansoni, the snails being present in three sites but absent from the other two sites. The objective of this study was to evaluate to what extent chemical and/or physical-morphological factors determine the distribution of B. pfeifferi between these five sites. It was found that none of the chemical constituents in the waters examined were outside the tolerance range of B. pfeifferi snails. Moreover, the composition of water from B. pfeifferi-free sites was not different from that in those sites where snails occurred. Furthermore, none of the physical-morphological constituents seemed likely to be a determinant for the absence of B. pfeifferi. In view of these findings, and those of previous studies, it is concluded that the focal distribution of B. pfeifferi cannot be associated with a single environmental factor and is rather the result of more complex interactions of habitat factors. PMID- 9332596 TI - Notes on the sand fly fauna (Diptera:Psychodidae) in the State of Rio Grande do Sul, Brazil. PMID- 9332598 TI - Limnomermis subtropicalis n. sp. (Nematoda:Mermithidae) a parasite of larvae of Simulium orbitale Lutz (Diptera:Simuliidae) in Argentina. AB - Limnomermis subtropicalis n. sp. (Nematoda:Mermithidae) a parasite of Simulium orbitale Lutz (Diptera:Simuliidae) found in Argentina is described and illustrated. This species is characterized by having medium sized amphids, pocket shaped, medium sized vagina, sculptured spicule, and by having 9 preanal, 7 postanal papillae in the ventral row, and 12 papillae in the lateral rows. PMID- 9332597 TI - Persistent infections by Leishmania (Viannia) braziliensis. AB - Here we review the phenomenon of persistency in Leishmania (Viannia) braziliensis infections. In other Leishmania species where appropriate animal models exist, considerable advances in the understanding of basic immunologic mechanisms of persistency have been made; for a review see Aebisher (1994). On the contrary, the evidences of persistence in infections with L. braziliensis rest on studies of human clinical cases many of which we summarized and discussed in this work. PMID- 9332599 TI - Characterization of Trypanosoma cruzi strains isolated from chronic chagasic patients, triatomines and opossums naturally infected from the State of Rio Grande do Sul, Brazil. AB - Thirty-five Trypanosoma cruzi strains were isolated from chronic chagasic patients, triatomines and opossums from different municipalities of the State of Rio Grande do Sul. Parasites were characterized by means of mice infectivity, enzyme electrophoresis and randomly amplified polymorphic DNA (RAPD) analysis. Twenty-nine strains were isolated from chagasic patients, 4 from triatomines (2 from Triatoma infestans and 2 from Panstrongylus megistus) and 2 from opossums Didelphis albiventris. Thirty-three T. cruzi strains were of low and 2 strains of high virulence in mice. Both virulent strains were isolated from P. megistus. Isoenzyme analysis of the strains showed 3 different zymodemes. Eleven strains isolated from chagasic patients and 2 from D. albiventris were Z2. Eighteen strains from patients and 2 from T. infestans were ZB and 2 T. cruzi strains isolated from P. megistus were Z1. RAPD profiles obtained with 4 random primers showed a high genetic heterogeneity of the T. cruzi strains. Zymodeme 2 and ZB strains were the more polymorphic. A band sharing analysis of the RAPD profiles of Z2 and ZB strains using 3 primers, showed a very low percentage of shared bands, 20% among 13 ZB strains and 14% among 13 Z2 strains. According to the isoenzyme results, 3 T. cruzi populations were present in State of Rio Grande do Sul. Zymodeme 2 and ZB strains were found infecting man (domiciliar transmission cycle) whereas Z1 strains were found infecting the sylvatic vector P. megistus. PMID- 9332600 TI - The type specimens of Apoidea (Hymenoptera) deposited in the Entomological Collection of Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. AB - A list of 41 Apoidea (Hymenoptera) type specimens deposited in the Entomological Collection of the Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil is presented. The types previously belonged to the Zikan and Schrottky private collections. A total of 11 holotypes and 30 paratypes are listed with their respective data and literature. PMID- 9332601 TI - Morphological and biometrical differences among Trypanosoma vivax isolates from Brazil and Bolivia. PMID- 9332602 TI - Paraorientatractis semiannulata n. g., n. sp. (Cosmocercoidea: Atractidae) from the large intestine of the side-necked turtle, Podocnemis unifilis Troschel, 1848 (Testudines: Pelomedusidae) in Brazil. AB - Specimens collected from the large intestine of the side-necked turtle Podocnemis unifilis Troschel, 1848 in the region of Cumina and Trombetas rivers near Para, Brazil are assigned to a new genus and new species of the nematode superfamily Cosmocercoidea and family Atractidae and named Paraorientatractis semiannulata. The new genus is separated from the nearest genus Orientatractis by the funnel shaped mouth opening, the presence of 4 distinct lips, 4 papillae in the internal cycle, one on each lip margin, 2 lateral amphids with large amphidial pores and absence of submedian papillae. It is also separated from Orientatractis and Proatractis by the presence of striated lateral alae which curve dorsally extending from mid oesophagus to mid tail, the difference in size of the vulvar opening and the presence of large transverse ridges or semiannules on the dorsal surface. The new species can be separated from the species of the genera Orientatractis and Proatractis by the characters that distinguish the genera and the arrangement of the caudal papillae on the male. A host/parasite list for Podocnemis spp. is included. PMID- 9332603 TI - Description of the immature stages of Anopheles (Nyssorhynchus) rondoni (Neiva & Pinto) (Diptera: Culicidae). off. AB - The larval and pupal stages of Anopheles (Nyssorhynchus) rondoni (Neiva and Pinto) (Albimanus Section) are fully described and illustrated for the first time. The larval stage is similar to An. (Nys.) strodei Root. It can be recognized by the following combination of characters: clypeal index, 1.6-2.9; single, aciculate setae 2,3-C; seta 3-C 0.5-0.7 the length of 2-C; setae 1,2-P rarely sharing a common tubercle; seta 1-P with narrow leaflets. The pupal stage is distinguished from other Nyssorhynchus by having setae 1-IV-VII and 5-V-VII branched. Only minor variation was found in setal counts between specimens from Peixoto de Azevedo, State of Mato Grosso and Bocaina, State of Sao Paulo, Brazil. PMID- 9332604 TI - Reproductive aspects of Haemaphysalis leporis-palustris. AB - The adult stage of Haemaphysalis leporis-palustris was studied. Two infestations consisted of male and female ticks on two rabbits (Oryctolagus cuniculus). Other two infestations consisted of only female ticks on two O. cuniculus. Females fed without males showed differences in some biological parameters when compared to females fed with males. Parthenogenesis is reported for the first time by one H. leporis-palustris female. PMID- 9332605 TI - On Leishmania enriettii and other enigmatic Leishmania species of the Neotropics. AB - There are 20 named species of the genus Leishmania at present recognized in the New World, of which 14 are known to infect man. The present paper discusses the biological, biochemical and ecological features, where known, of six species which have not till now been found to cause human leishmaniasis; namely, Leishmania (Leishmania) enriettii, L. (L.) hertigi, L. (L.) deanei, L. (L.) aristidesi, L. (L.) forattinii and L. (Viannia) equatorensis. A protocol is suggested for attempts to discover the natural mammalian host(s) and sandfly vector of L. (L.) enriettii. Doubt is cast on the validity of the species L. herreri, described in Costa Rican sloths. Following the concensus of opinion that modern trypanosomatids derive from monogenetic intestinal flagellates of arthropods, phlebotomine sandflies are best regarded as the primary hosts of Leishmania species, with mammals acting as secondary hosts providing a source of parasites for these insects. There are probably natural barriers limiting the life-cycle of most leishmanial parasites to specific sandfly vectors. PMID- 9332606 TI - Analysis of respiratory syncytial virus in clinical samples by reverse transcriptase-polymerase chain reaction restriction mapping. AB - The aim of this study was to develop a polymerase chain reaction (PCR) for the detection of respiratory syncytial virus (RSV) genomes. The primers were designed from published sequences and selected from conserved regions of the genome encoding for the N protein of subgroups A and B of RSV. PCR was applied to 20 specimens from children admitted to the respiratory ward of "William Soler" Pediatric Hospital in Havana City with a clinical diagnosis of bronchiolitis. The PCR was compared with viral isolation and with an indirect immunofluorescence technique that employs monoclonal antibodies of subgroups A and B. Of 20 nasopharyngeal exudates, 10 were found positive by the three assayed methods. In only two cases, samples that yielded positive RNA-PCR were found negative by indirect immunofluorescence and cell culture. Considering viral isolation as the "gold standard" technique, RNA-PCR had 100% sensitivity and 80% specificity. RNA PCR is a specific and sensitive technique for the detection of the RSV genome. Technical advantages are discussed. PMID- 9332608 TI - Inexpensive alternative material for the isolation of larvae with the Baermann method. PMID- 9332609 TI - Genetic variability and microdistribution of Triatoma infestans genotypes and Trypanosoma cruzi clones in Arequipa region (Peru). AB - The genetic variability of Triatoma infestans and Trypanosoma cruzi populations was studied by isoenzyme analysis in two distinct areas of Arequipa province (Peru); one, Santa Rita de Siguas, being an endemic area for Chagas' disease, the second, Arequipa, recently infected. Analysis of T. infestans genetic variability indicates, (i) temporal stability of genotypes found in Santa Rita de Siguas, (ii) high genetic differences between Arequipa and Santa Rita de Siguas populations suggesting minor contact between them, (iii) multiple origin of the T. infestans population in Arequipa, and (iv) poor dispersal capacity of T. infestans: the panmictic unit could be reduce to a house. Parasite isoenzyme analysis was performed in 29 Peruvian stocks of T. cruzi, mainly isolated from bugs taken in a single locality, Santa Rita de Siguas. The results show, (i) a high genetic polymorphism, (ii) nine different multilocus genotypes were detected and clustered in two different clades, (iii) most of the parasite isolates pertained to one of the clade and were genetically similar to those analyzed 12 years before. This sample allowed the study of the mating system of T. cruzi in strict sympathic conditions and gave more strength to the hypothesis of the clonal structure of T. cruzi populations. PMID- 9332607 TI - A simple reverse transcription-polymerase chain reaction for dengue type 2 virus identification. AB - We show here a simplified reverse transcription-polymerase chain reaction (RT PCR) for identification of dengue type 2 virus. Three dengue type 2 virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD, as a negative control, were used in this study. C6/36 cells were infected with the virus, and tissue culture fluids were collected after 7 days of infection period. The RT-PCR, a combination of RT and PCR done after a single addition of reagents in a single reaction vessel was carried out following a digestion of virus with 1% Nonidet P-40. The 50 microliters assay reaction mixture included 50 pmol of a dengue type 2 specific primer pair amplifying a 210 base pair sequence of the envelope protein gene, 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, and IU of thermostable Taq DNA polymerase. The reagent mixture was incubated for 15 min at 37 degrees C for RT followed by a variable amount of cycles of two-step PCR amplification (92 degrees C for 60 sec, 53 degrees C for 60 sec) with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized with UV light after gel incubation in ethidium bromide solution. DNA bands were observed after 25 and 30 cycles of PCR. Virus amount as low as 10(2.8) TCID 50/ml was detected by RT PCR. Specific DNA amplification was observed with the three dengue type 2 strains. This assay has advantages compared to other RT-PCRs: it avoids laborious extraction of virus RNA; the combination of RT and PCR reduces assay time, facilitates the performance and reduces risk of contamination; the two-step PCR cycle produces a clear DNA amplification, saves assay time and simplifies the technique. PMID- 9332610 TI - Contrasting genomic variability between clones from field isolates and laboratory populations of Schistosoma mansoni. AB - The extent of genomic variability of clones of Schistosoma mansoni obtained from field isolates was compared with that of strains that have been laboratory maintained. Analysis was undertaken using randomly amplified polymorphic DNAs (RAPDs) generated with three primers. Phenograms showing the similarity among the clones were constructed. The data showed that while the laboratory strain is highly homogeneous the clones derived from the field populations were highly variable with 43% of RAPDs exhibiting polymorphisms among 23 clones. Clones isolated from the same infected individual were always more closely grouped than clones from different individuals. The data clearly demonstrated that earlier analyses of the genomic variability in S. mansoni have underestimated this phenomenon due to the failure to examine field isolates. PMID- 9332611 TI - Conjugation by mosquito pathogenic strains of Bacillus sphaericus. AB - A mosquito pathogenic strain of Bacillus sphaericus carried out the conjugal transfer of plasmid pAM beta 1 to other strains of its own and two other serotypes. However, it was unable to conjugate with mosquito pathogens from three other serotypes, with B. sphaericus of other DNA homology groups or with three other species of Bacillus. Conjugation frequency was highest with a strain having an altered surface layer (S layer). Conjugal transfer of pAM beta 1 was not detected in mosquito larval cadavers. B. sphaericus 2362 was unable to mobilize pUB110 for transfer to strains that had served as recipients of pAM beta 1. These observations suggest that it is unlikely that genetically engineered B. sphaericus carrying a recombinant plasmid could pass that plasmid to other bacteria. PMID- 9332612 TI - A comparative study of the organic acid content of the hemolymph of Schistosoma mansoni-resistant and susceptible strains of Biomphalaria glabrata. AB - The freshwater snail Biomphalaria glabrata is an intermediate host of the trematode Schistosoma mansoni. However, some strains of B. glabrata are resistant to successful infection by S. mansoni larvae. The present work examines the profile of organic acids present in S. mansoni-resistant and -susceptible strains of B. glabrata, in order to determine whether the type of organic acid present is related to susceptibility. The organic acids were extracted from the hemolymph of two susceptible B. glabrata strains (PR, Puerto Rico and Ba, Jacobina-Bahia from Brazil), and from the resistant strains 13-16-R1 and 10R2, using solid phase extraction procedures followed by high performance liquid chromatography. The organic acids obtained were analyzed and identified by comparison with known standards. Pyruvate, lactate, succinate, malate, fumarate, acetate, propionate, beta-hydroxybutyrate and acetoacetate were detected in all hemolymph samples. Under standard conditions, the concentration of each of these substances varied among the strains tested and appeared to be specific for each strain. An interesting variation was the low concentration of pyruvate in the hemolymph of PR-snails. Only the concentration of fumarate was consistently different (p < or = 0.05) between resistant and susceptible strains. PMID- 9332613 TI - Effects of goyazensolide during in vitro cultivation of Schistosoma mansoni. AB - Goyazensolide, a component extracted of Eremanthus goyazensis showed a significant inhibitory effect on egg-laying of Schistosoma mansoni during in vitro cultivation of this parasite. Motility of the worms was also reduced under treatment with goyazensolide and 90% of mortality was reached with concentrations up to 4 micrograms/ml. It has found that separated worms were more susceptible than worms pairing during drug exposition and female alone was significantly more susceptible than male worm in the same conditions of in vitro cultivation. Natural products isolated from plants represent potential sources for the identification of structures useful for the design of alternative molecules to be used as new drug substances against several infectious diseases. PMID- 9332614 TI - Incoordination, paralysis and recovery after pyrethroid treatment on nymphs III of Triatoma infestans (Hemiptera: Reduviidae). AB - Symptoms of poisoning for deltamethrin and cis-permethrin on nymphs III of Triatoma infestans were described. The time required for incoordination and paralysis were determined. Deltamethrin was equal or more rapid in the onset of the first effect (accordingly to dose), and cis-permethrin in the onset of the second one. There were no significant differences between incoordination doses 50% (IncD50s) at different times for the two pyrethroids. They showed equivalent incoordination power, but the nymphs treated with deltamethrin recovered slightly more rapid and in greater amount than the nymphs treated with cis-permethrin. The recovery was inhibited by the simultaneous application of piperonyl butoxide. This result suggests that biotransformation by mixed-function microsomal oxidases are involved in the process of recovery. PMID- 9332616 TI - [Growth, food conversion and mortality in Eretmochelys imbricata (Reptilia: Chelonidae) in artificial ponds in Costa Rica]. AB - Growth rates, feed conversion and mortality of cultured Eretmochelys imbricata (hawksbill turtle) were studied in concrete raceways by feeding with fresh fish meal (tilapia) and in duplicate (tanks of 21 m2). The turtles were 11 months old at the beginning of the experiment, with and average caparace straight length of 23.64 +/- 1.94 cm, an average caparace curved length of 24.15 +/- 1.94 cm and an average weight of 1527 +/- 2.54 g. The experiment was carried out during six months and a density of 3 ind/m2 was used. The equation of Von Bertalanffy for the growth of E. imbricata was LRC = 3.5 + [(82.0 -3.5) (1-e-0.67432 (t))]. The relationship between caparace straight length-weight was W = 5.207 x 10(-3) LRC 3.8807 (r = 0.99). The feed conversion was 1.54 +/- 0.74 and the mortality was zero during the study. PMID- 9332615 TI - [Embryonic mortality and hatching rate of Trachemys scripta (Testudines: Emydidae) eggs artificially grown in a protected natural area]. AB - In the northeast of Costa Rica, illegal collecting, habitat destruction and heavy predation on adult females, young and eggs of Trachemys scripta produced a population decline in recent years. The slider turtle has a high reproductive potential and nests were relocated. Between January and March 1991, 179 nests (3220 eggs) were collected from natural areas of Cano Negro National Wildlife Refuge, Costa Rica and relocated for incubation under natural conditions in a protected site. The mean of number of eggs per clutch was 19.5 +/- 4.5 (range 8 to 31) and the mean incubation period was 77.4 +/- 11.4 days (range 50 to 110 days). The mean hatching success was 89.6 +/- 3.4% (range 60.0 to 100%). The mean early and mid embryonic mortality was 5.5 +/- 4.3%, the average late embryonic mortality was 8.5 +/- 3.5% and infertility 1.8 +/- 1.0%. PMID- 9332617 TI - Reproduction and conservation of the leatherback turtle Dermochelys coriacea (Testudines: Dermochelyidae) in Gandoca, Costa Rica. AB - The leatherback turtle was studied in Gandoca, an important nesting beach on the southeastern Caribbean coast of Costa Rica (82 degrees 37' W; 09 degrees 37' N). In 1994, a total of 530 nests was recorded during the nesting season (February/July) and 160 leatherbacks were tagged; five were remigrants from the 1992 season and 15 carried tags from elsewhere. Eighty eight females only nested once. Mean curve carapace measurements were length 153.8 cm and width 112.0 cm. A hatchery received 82 clutches, with 6277 normal eggs. Their mean incubation period was 62.24 days (range: 56-68 days). Average hatching rate was 55.10% (S.D.: 25.04, range 15-96%). Extensive erosion, beach debris and poaching activity represent the main hazards for nesting in Gandoca. PMID- 9332618 TI - [Aggressive behavior of the male volcano mouse Neotomodon alstoni (Rodentia: Cricetidae)]. AB - A study was carried out on the agonistic behavior of the male volcano mouse, Neotomodon alstoni, with 50 pairs of males which were classified as possible dominants (D) and subordinates (S), utilizing Melzack-Thompson's Method. The aggressiveness levels exhibited by this mouse were recording in the combinations: D vs. D and S vs. S. Two groups were formed: Group I with 12 pairs of D males and 13 of S males, and Group II with 11 pairs of D males and 14 of S males. In Group I the aggressiveness level was quantified after one week of mating and after another week of isolation, and in Group II the sequence of observation was inverted. The aggressiveness level was measured by the number of attacks per hour, an attack being defined as the aggressive physical contac of an animal (aggressor) with another (attacked). The kinds of behavior registered, including offense, defense, and submission patterns, revealed hierarchic relationships. Dominance was correlated significatively (p < 0.05) with a higher level of aggressiveness. PMID- 9332619 TI - Natural presence of the bacterium Salmonella sp. in hen eggs consumed in Costa Rica. PMID- 9332620 TI - Sailing life's uncharted seas: advance directives and emergency medicine patients. PMID- 9332621 TI - Flumazenil: a pharmacologic antidote with limited medical toxicology utility, or ... an antidote in search of an overdose. PMID- 9332622 TI - A significant difference is one that makes a difference. PMID- 9332623 TI - Pilot instrument proficiency: flight of fancy or emergency medical services concern? PMID- 9332624 TI - Computerized simulation technology for clinical teaching and testing. PMID- 9332625 TI - Safety and efficacy of flumazenil in reversing conscious sedation in the emergency department. Emergency Medicine Conscious Sedation Study Group. AB - OBJECTIVES: To evaluate the safety and efficacy of flumazenil vs placebo in reversing fentanyl and midazolam-induced conscious sedation in ED patients undergoing a short, painful procedure. METHODS: This was a multicenter, randomized, parallel, double-blind, placebo-controlled study conducted at 9 university-affiliated teaching hospitals. Patients > 18 years of age requiring conscious sedation for a painful procedure expected to last < 20 minutes were eligible for inclusion in the study. Patients received 2 micrograms/kg of fentanyl, followed by midazolam titrated to the desired level of sedation. Patients were then randomized to receive either flumazenil or placebo in a 3:1 ratio (flumazenil:placebo). Vital signs, O2 saturation, and alertness were recorded at regular intervals. Prior to ED release, patients were asked to rate the amount of discomfort they experienced and the level of relaxation achieved on a 10-cm visual analog scale (VAS). They also were questioned about their recall for the procedure and satisfaction with the drug regimen. Physicians also rated their satisfaction with the drug regimen on a 10-cm VAS. RESULTS: Overall, 133 patients received flumazenil and 46 patients received placebo. Patients in the 2 groups received similar doses of midazolam. The patients who received flumazenil returned to baseline alertness earlier (11.1 min vs 24.8 min, p < 0.001) and at a faster rate than did the patients given placebo. Actual intervals from procedure completion until release from the ED did not differ between the 2 groups (98.2 +/ 3.6 min flumazenil vs 96.9 +/- 5.8 min placebo; p = 0.89). The amounts of discomfort experienced, levels of relaxation achieved, recalls for the procedure, and both patient and physician satisfactions were also similar for the 2 groups. There were no serious adverse effects related to the study drug, and minor adverse effects were similar for the 2 groups. CONCLUSION: Flumazenil is safe and efficacious in reversing midazolam-induced sedation in ED patients given a combination of fentanyl and midazolam to facilitate the performance of a short, painful procedure. The patients given flumazenil returned to baseline alertness earlier and at a faster rate than did the patients given placebo. However, flumazenil did not alter the actual interval from procedure completion until ED release. PMID- 9332626 TI - Rotating shiftwork schedules: can we enhance physician adaptation to night shifts? AB - OBJECTIVES: To evaluate the effectiveness of a broad, literature-based night shiftwork intervention for enhancement of emergency physicians' (EPs') adaptation to night rotations. METHODS: A prospective, double-blind, active placebo controlled study was conducted on 6 attending physicians in a university hospital ED. Three data sets were collected under the following conditions: baseline, after active placebo intervention, and after experimental intervention. In each condition, data were collected when the physicians worked both night and day shifts. Measurements included ambulatory polysomnographic recordings of the main sleep periods, objective performance tests administered several times during the subjects' shifts, and daily subjective ratings of the subjects' sleep, moods, and intervention use. RESULTS: The subjects slept an average of 5 hr 42 min across all conditions. After night shifts, the subjects slept significantly less than they did after day shifts (5 hr 13 min vs 6 hr 20 min; p < 0.05). The physicians' vigilance reaction times and times for intubation of a mannequin were significantly slower during night shifts than they were during day shifts (p = 0.007 and p < 0.04, respectively), but performances on ECG analysis did not significantly differ between night and day shifts. Mood ratings were significantly more negative during night shifts than they were during day shifts (more sluggish p < 0.04, less motivated p < 0.03, and less clear thinking p < 0.04). The strategies in the experimental intervention were used 85% of the time according to logbook entries. The experimental and active placebo interventions did not significantly improve the physician's performance, or mood on the night shift, although the subjects slept more after both interventions. CONCLUSIONS: Although the experimental intervention was successfully implemented, it failed to significantly improve attending physicians' sleep, performance, or mood on night shifts. A decrease in speed of intubation, vigilance reaction times, and subjective alertness was evident each time the physicians rotated through the night shift. These findings plus the limited sleep across all conditions and shifts suggest that circadian-mediated disruptions of waking neurobehavioral functions and sleep deprivation are problems in EPs. PMID- 9332628 TI - Normal intrauterine pregnancy is unlikely in emergency department patients with either menstrual days > 38 days or beta-hCG > 3,000 mIU/mL, but without a gestational sac on ultrasonography. AB - OBJECTIVE: To determine whether the absence of a gestational sac on transvaginal ultrasonography in patients with a quantitative beta-human chorionic gonadotropin (beta-hCG) > 3,000 mIU/mL and/or menstrual days > 38 excludes the diagnosis of a normal intrauterine pregnancy (IUP). METHODS: A retrospective analysis was performed of ED patients evaluated from August 1991 to December 1994 at an urban teaching hospital. Patients presented with abdominal pain and/or vaginal bleeding and had a positive serum beta-hCG test. Patients who had transvaginal ultrasonographies performed during the ED visit that were read as indeterminate were reviewed. Menstrual days were determined by subtracting the date of the last normal menstrual period (LMP) from the ED visit date. ED beta-hCGs were quantified. Patients were excluded if the LMP or quantitative beta-hCG result was not available or if the final diagnosis could not be definitively determined. RESULTS: 248 patients met eligibility criteria; of these, 54 were excluded. Therefore, 194 patients were enrolled. Menstrual days ranged from 5 to 151, with a median of 54 days. Of 143 patients with menstrual days > 38 and no gestational sac by ultrasonography, only 4 (2.8%) had a final diagnosis of normal IUP. The menstrual days for the 4 normal IUPs were 39, 41, 42, and 59 days. beta-hCGs ranged from 19 to 151,926 mIU/mL, with a median of 2,410 mIU/mL. None of the 74 patients with a beta-hCG > 3,000 mIU/mL and no gestational sac by ultrasonography had a final diagnosis of normal IUP. CONCLUSION: In patients with either a beta hCG > 3,000 mIU/mL or menstrual days > 38 and no gestational sac by transvaginal ultrasonography, the likelihood of a normal IUP is low. PMID- 9332627 TI - Underrecognition of cervical Neisseria gonorrhoeae and Chlamydia trachomatis infections in the emergency department. AB - OBJECTIVES: 1) To quantify the frequency of underrecognized Neisseria gonorrhoeae and Chlamydia trachomatis cervical infections in women tested in the ED, 2) to describe and compare the characteristics of those treated and not treated during the initial visit, and 3) to quantify the delay interval until treatment was provided. METHODS: A 2-year, retrospective consecutive case series was performed from June 1, 1992, to May 31, 1994. There were 148 women with > or = 1 discrete occurrence of culture-proven cervical N. gonorrhoeae or C. trachomatis infection studied. All the patients were evaluated in a university-affiliated, tertiary care hospital-based ED with a large rural referral area. The main outcome measures were the proportions of patients with positive cultures both treated and not treated in the ED, the clinical characteristics of each group, and the proportion remaining untreated or experiencing treatment delays of > 2 weeks after attempted phone, mail, and public health follow-up. RESULTS: Of 157 occurrences of positive cultures for N. gonorrhoeae or C. trachomatis, 86 (53%) were treated with a regimen suggested by the CDC prior to ED release. The proportion of women with isolated C. trachomatis infections that were underrecognized and untreated initially was larger than the proportions with isolated N. gonorrhoeae or combined infections (79% vs 27% and 53%, respectively, p < 0.0001). Women with findings suggestive of advanced disease (history of fever or chills, or examination evidence of temperature > 38 degrees C, purulent vaginal discharge, or any uterine/salpinx/ovarian tenderness) were more often recognized and treated with appropriate antibiotics initially (p = 0.02 to < 0.00001 for all). After phone, mail, and public health follow-up, treatment could not be documented for 25% of the occurrences, in all cases due to an inability to locate the patient. An additional 20% of the women did not receive treatment for 14-60 days. CONCLUSIONS: In this population, both N. gonorrhoeae and C. trachomatis cervical infections are frequently underrecognized in the ED, with isolated C. trachomatis infections associated with significantly higher proportions of underrecognition. Many affected women remain untreated for extended intervals, creating public and individual health risks. Improved point of contact detection, follow-up, and treatment policies are needed to limit these risks. PMID- 9332629 TI - Role of pilot instrument proficiency in the safety of helicopter emergency medical services. AB - OBJECTIVES: To determine whether instrument-proficient pilots would more safely manage a flight into unplanned instrument meteorologic conditions (IMC) than would nonproficient pilots. METHODS: A controlled experimental study was performed using a full-motion helicopter simulator. Participants were emergency medical services (EMS) pilots with commercial licenses and previous simulator experience who were blinded to the study design and hypothesis. During a simulated EMS mission, cloud ceiling and visibility were decreased until IMC prevailed, and pilot actions were recorded. Data included the altitude at which the aircraft entered IMC, and whether the pilots maintained control of the aircraft, flew within aviation standards (i.e., bank angle, airspeed), and safely landed. RESULTS: Twenty-eight pilots (13 instrument-proficient, 15 nonproficient) participated; they had a median of 6,300 hours of helicopter experience. Two pilots crashed, both from the nonproficient group. The instrument-proficient pilots lost control less often (15% vs 67%, p < 0.05), maintained instrument standards more often (77% vs 40%, p < 0.05), and entered IMC at a higher altitude (689 feet vs 517 feet, p < 0.05) compared with the nonproficient pilots. Instructor comments indicated that the nonproficient pilots made more errors than did the instrument-proficient pilots. CONCLUSIONS: Instrument-proficient pilots more safely manage an unexpected encounter with IMC. Helicopter EMS programs should strongly consider maintaining instrument proficiency to enhance safety. PMID- 9332630 TI - Are emergency department patients thinking about advance directives? AB - OBJECTIVES: To assess the percentage of adult patients presenting to an urban ED who have a written advance directive (AD) and to determine whether age, sex, a patient's perception of his or her health status, and having a regular physician are associated with the patient's having an AD. METHODS: This was a cross sectional patient survey performed at a community teaching hospital ED. Surveys were completed by 511 adult ED patients during representative shifts over a 3 month period. The questions included age, sex, "self-reported" health status, whether the patient had a "regular" physician, a patient-generated list of medical problems, and whether the patient had a written AD. For this study, ADs included health care proxies, living wills, and do-not-attempt-resuscitation (DNAR) orders. RESULTS: Of the patients surveyed, 27% reported having an AD. Males and females were equally likely to have an AD. Factors associated with an increased likelihood of having an AD were older age, having a "regular" physician, and the patient's perception of his or her health status as ill. Most patients who had an AD (82%) discussed it with their families, but only 48% discussed it with their physicians. CONCLUSION: Only 27% of the adult patients presenting to the ED had an AD. Older age, the patient's perception of his or her health status as ill, and having a "regular" physician increased the likelihood of having an AD. PMID- 9332631 TI - Assessing bedside cardiologic examination skills using "Harvey," a cardiology patient simulator. AB - OBJECTIVE: To assess the cardiovascular physical examination skills of emergency medicine (EM) housestaff and attending physicians. METHODS: Prospective, cohort assessment of EM housestaff and faculty performance on 3 valvular abnormality simulations (mitral regurgitation, mitral stenosis, and aortic regurgitation) conducted on the cardiology patient simulator, "Harvey." Participants examined each of the 3 study disease simulations and proposed a diagnosis (session I). They were then given a cardiac examination form and repeated the programmed simulations (session II). The examination form was used to prompt physicians to interpret 23 separate cardiac findings for each simulation in a multiple-choice format. RESULTS: Forty-six EM housestaff (PGY1-3) and attending physicians were tested over a 2-month study period. Physician responses did not differ significantly among the different levels of postgraduate training. The overall correct response rates for participants were 59% for aortic regurgitation, 48% for mitral regurgitation, and 17% for mitral stenosis. For aortic regurgitation, recognition of a widened pulse pressure and recognition of diastolic decrescendo murmur were associated with a correct diagnosis (p < 0.01). For mitral regurgitation, correct assessment of the contour of the holosystolic murmur predicted a correct diagnosis (p < 0.001). For mitral stenosis, proper characterization of the mitral area diastolic murmur predicted a correct diagnosis (p < 0.001). CONCLUSION: Housestaff and faculty had difficulty establishing a correct diagnosis for simulations of 3 common valvular heart diseases. However, accurate recognition of a few critical signs was associated with a correct diagnosis in each simulation. Training programs may need to focus attention on selected key components of the cardiovascular examination to facilitate teaching of physical diagnosis. PMID- 9332632 TI - Early stroke recognition: developing an out-of-hospital NIH Stroke Scale. AB - OBJECTIVE: To develop an abbreviated and practical neurologic scale that could assist emergency medical services or triage personnel in identifying patients with stroke. METHODS: A prospective, observational, cohort study was performed at university-based EDs. Participants were 74 patients treated in a thrombolytic stroke trial and 225 consecutive non-stroke patients evaluated during 4 random 12 hour shifts in the ED. Scores on the NIH Stroke Scale were obtained for all patients by physicians. Items of this scale were modified and recoded to a binomial (normal or abnormal) scale. Serial univariate analyses using chi 2 were performed to rank items. Recursive partitioning was then performed to develop the decision rule for predicting the presence of stroke. RESULTS: Three items identified 100% of patients with stroke: facial palsy, motor arm, and dysarthria. An Abbreviated NIH Stroke Scale based on these items had a sensitivity of 100% and a specificity of 92%. A proposed Out-of-hospital NIH Stroke Scale consisting of facial palsy, motor arm, and a combination of dysarthria and best language items (abnormal speech) had a sensitivity of 100% and a specificity of 88%. CONCLUSION: Using the derivation data set, a proposed Out-of-hospital NIH Stroke Scale had a high sensitivity and specificity for identifying patients with stroke when performed by physicians in this group of 299 ED patients. Prospective studies of other health care professionals using the scale in the out-of-hospital arena are needed. PMID- 9332633 TI - Vertebral artery dissection. AB - Vertebral artery dissections (VADs) following a variety of minor traumatic mechanisms have been previously reported. This article reports 2 cases of VAD with delayed recognition following motor vehicle collisions (MVCs). The first VAD patient developed major neurologic abnormalities 28 hours after an MVC. The second VAD patient presented with 3 weeks of neck and head pain beginning 8 weeks after an MVC and subsequent chiropractic manipulation. The anatomy and pathophysiology of VAD are reviewed. Early ED recognition prior to the onset of major neurologic deficits (e.g., paresis, dysarthria, ataxia, or altered mental status) is emphasized. An algorithm for the ED management of the entity is suggested. PMID- 9332635 TI - Cricothyrotomy concern. PMID- 9332634 TI - The development of international emergency medicine: a role for U.S. emergency physicians and organizations. SAEM International Interest Group. AB - There is a rapidly growing interest in emergency medicine (EM) and emergency out of-hospital care throughout the world. In most countries, the specialty of EM is either nonexistent or in an early stage of development. Many countries have recognized the need for, and value of, establishing a quality emergency health care system and are striving to create the specialty. These systems do not have to be high tech and expense but can focus on providing appropriate emergency training to physicians and other health care workers. Rather than repeatedly "reinventing the wheel" with the start of each new emergency care system, the preexisting knowledge base of EM can be shared with these countries. Since the United States has an advanced emergency health care system and the longest history of recognizing EM as a distinct medical specialty, lessons learned in the United States may benefit other countries. In order to provide appropriate advice to countries in the early phase of emergency health care development, careful assessment of national resources, governmental structure, population demographics, culture, and health care needs is necessary. This paper lists specific recommendations for EM organizations and physicians seeking to assist the development of the specialty of EM internationally. PMID- 9332636 TI - Unusual lubricants do not interfere with the Hemoccult test for occult blood. PMID- 9332637 TI - Potential role of rural scene air medical transport of patients with chest pain. PMID- 9332638 TI - Concerns regarding the welfare check distribution study. PMID- 9332639 TI - Concerns regarding the welfare check distribution study. PMID- 9332640 TI - Methemoglobinemia from a topical oral anesthetic. PMID- 9332650 TI - Multiple malformation syndrome following fluconazole use in pregnancy: report of an additional patient. PMID- 9332651 TI - Xq28-linked noncompaction of the left ventricular myocardium: prenatal diagnosis and pathologic analysis of affected individuals. AB - Isolated noncompaction of the left ventricular myocardium (INVM) is characterized by the presence of numerous prominent trabeculations and deep intertrabecular recesses within the left ventricle, sometimes also affecting the right ventricle and interventricular septum. Familial occurrence of this disorder was described previously. We present a family in which 6 affected individuals demonstrated X linked recessive inheritance of this trait. Affected relatives presented postnatally with left ventricular failure and arrhythmias, associated with the pathognomonic echocardiographic findings of INVM. The usual findings of Barth syndrome (neutropenia, growth retardation, elevated urinary organic acids, low carnitine levels, and mitochondrial abnormalities) were either absent or found inconsistently. Fetal echocardiograms obtained between 24-30 weeks of gestation in 3 of the affected males showed a dilated left ventricle in one heart, but were not otherwise diagnostic of INVM in any of the cases. Four of the affected individuals died during infancy, one is in cardiac failure at age 8 months, and one is alive following cardiac transplant at age 9 months. The hearts from infants who died or underwent transplantation appeared, on gross examination, to be enlarged, with coarse, deep ventricular trabeculations and prominent endocardial fibroelastosis. Histologically, there were loosely organized fascicles of myocytes in subepicardial and midmyocardial zones of both ventricles, and the myocytes showed thin, often angulated fibers with prominent central clearing and reduced numbers of filaments. Markedly elongated mitochondria were present in some ventricular myocytes from one specimen, but this finding was not reproducible. Genetic linkage analysis has localized INVM to the Xq28 region, where other myopathies with cardiac involvement have been located. PMID- 9332652 TI - Craniofacial structure in diastrophic dysplasia--a cephalometric study. AB - Diastrophic dysplasia (DTD) is a well characterized, recessively inherited osteochondrodysplasia. Forty-eight patients with DTD were studied for craniofacial characteristics. Of these patients, 58% had cleft palate. A cephalometric analysis based on lateral cephalograms was performed. We observed a short anterior cranial base, vertical nasal bones, short and posteriorly positioned upper and lower jaws, increased anterior facial height, increase in the sagittal length of the body of the cervical vertebrae, and an abnormal dens of the second cervical vertebra. DTDST, in which mutations responsible for the disease occur, is a gene that codes for a sulphate transporter membrane protein. The craniofacial anomalies in DTD most likely result from deficient development and growth of cartilaginous structures and are probably due to defective sulfation of the proteoglycans of the cartilage. PMID- 9332653 TI - Discriminant analysis of the Ullrich-Turner syndrome neurocognitive profile. AB - Ullrich-Turner syndrome (UTS), or monosomy X, is a genetic disorder characterized by short stature, gonadal dysgenesis, and a particular neurocognitive profile of normally developed language abilities (particularly verbal IQ) and impaired visual-spatial and/or visual-perceptual abilities. The most frequently described profile in UTS includes difficulty with tasks involving memory and attention, decreased arithmetic skills, and impaired visual spatial processing. We used discriminant function analysis (DFA) to distinguish between the neurocognitive profiles of girls with UTS vs. controls matched for age, height, IQ, and socioeconomic status. DFA is a statistical method for deriving a linear function that optimally weights parameters to permit sensitive and specific differentiation among groups. We developed a modified discriminant function, based on seven cognitive test scores, that successfully discriminated between the UTS and control subjects with a sensitivity of 0.45 and a specificity of 0.97. To validate its performance, we applied the discriminant function to a small group of 45,X UTS subjects (n = 13) and control female subjects (n = 25), ages 7-16 years, who were not part of the previous analyses. The discriminant function (DF) identified 54% of these 13 UTS subjects as having the "UTS neurocognitive profile" and 92% of the 25 control subjects as not having the profile. We also compared the DF scores of UTS girls with various mosaic karyotypes and found that the group with 46,XX mosaicism had significantly higher scores (i.e., closer to normal controls) than the other two mosaic groups (t = 2.86, P < 0.005). The results of this study should be useful for genetic counseling and planning educational programs for girls with UTS. PMID- 9332654 TI - Ring chromosome 4 mosaicism coincidence of oligomeganephronia and signs of Seckel syndrome. AB - We present a patient with features suggestive of Seckel syndrome who was found to be mosaic for ring 4 chromosome. Seckel syndrome is a rare entity characterized by marked growth retardation, microcephaly, facies characterized by receding forehead and chin, large beaked nose, and severe retardation, usually thought to be inherited as an autosomal recessive condition. In addition, our patient had oligomeganephronia, a rare and usually sporadic renal malformation, previously reported in two other patients with abnormalities of chromosome 4. Besides pointing out the overlap between the Seckel phenotype and Wolf-Hirschhorn syndrome, our patient illustrates the need to consider cytogenetic studies in patients with the Seckel phenotype, so that accurate diagnoses can be given to families. Also, the case suggests that there may be a locus for oligomeganephronia distal to the Wolf-Hirschhorn critical region on 4p. PMID- 9332655 TI - Glycogen storage disease type 1a in Israel: biochemical, clinical, and mutational studies. AB - Glycogen storage disease type 1a (von Gierke disease, GSD 1a) is caused by the deficiency of microsomal glucose-6-phosphatase (G6Pase) activity which catalyzes the final common step of glycogenolysis and gluconeogenesis. The recent cloning of the G6Pase cDNA and characterization of the human G6Pase gene enabled the characterization of the mutations causing GSD 1a. This, in turn, allows the introduction of a noninvasive DNA-based diagnosis that provides reliable carrier testing and prenatal diagnosis. In this study, we report the biochemical and clinical characteristics as well as mutational analyses of 12 Israeli GSD 1a patients of different families, who represent most GSD 1a patients in Israel. The mutations, G6Pase activity, and glycogen content of 7 of these patients were reported previously. The biochemical data and clinical findings of all patients were similar and compatible with those described in other reports. All 9 Jewish patients, as well as one Muslim Arab patient, presented the R83C mutation. Two Muslim Arab patients had the V166G mutation which was not found in other patients' populations. The V166G mutation, which was introduced into the G6Pase cDNA by site-directed mutagenesis following transient expression in COS-1 cells, was shown to cause complete inactivation of the G6Pase. The characterization of all GSD 1a mutations in the Israeli population lends itself to carrier testing in these families as well as to prenatal diagnosis, which was carried out in 2 families. Since all Ashkenzai Jewish patients harbor the same mutation, our study suggests that DNA-based diagnosis may be used as an initial diagnostic step in Ashkenazi Jews suspected of having GSD 1a, thereby avoiding liver biopsy. PMID- 9332656 TI - Neurosonographic diagnosis of thalamic/basal ganglia vasculopathy in trisomy 13- an important diagnostic aid. AB - We performed a retrospective review of all the infants diagnosed with trisomy 13 in our institution from 1982 to 1995 and evaluated the neurosonographic findings along with their clinical information and cytogenetic analysis. Nine babies were admitted with trisomy 13. Sonography of the head was performed on 4 patients, and demonstrated in all of them a linear, branching, echogenic pattern in the thalamus/basal ganglia. Doppler evaluation of the thalamus/basal ganglia was performed in 3 of the 4 cases and confirmed these linear echogenicities to be of vascular origin. This is the first study to evaluated the occurrence of this finding in a specific syndrome, namely trisomy 13. PMID- 9332657 TI - Familial occurrence of pulmonary atresia with intact ventricular septum. AB - Pulmonary atresia with intact ventricular septum (PA/IVS) is a rare disease, accounting for less than 3% of all congenital heart lesions. The cause of PA/IVS is unknown. We report the occurrence of two first cousins with PA/IVS, suggestive of autosomal dominant inheritance with incomplete penetrance. The study of such families should ultimately lead to the identification of the gene(s) that cause congenital heart disease. PMID- 9332658 TI - Cytogenetic analysis of spontaneous abortions: comparison of techniques and assessment of the incidence of confined placental mosaicism. AB - Cytogenetic studies on spontaneous abortions traditionally have used one of two methodologies, direct preparations or long-term culture, to determine the chromosome constitution of either the cytotrophoblast or villous stroma, respectively. Few studies have utilized both techniques simultaneously to compare the relative efficiencies of each method and to assess the contribution of confined placental mosaicism (CPM). The present report summarizes cytogenetic studies on 691 consecutive spontaneous abortions using long-term culture, direct preparations, or both. All 691 cases were analyzed by long-term culture and 177 cases were analyzed using both long-term culture and direct preparations. The results indicate that the two methods have similar success rates, 82% for long term culture and 76% for direct preparation; however, the proportion of normal females was significantly increased in the culture method, presumably attributable to maternal contamination. In 107 cases, results were obtained from both methods with 22 discrepancies identified. However, most of these involved a 46,XX result in culture, consistent with maternal contamination in the cultured preparation. Therefore, to estimate the proportion of CPM we excluded cases with a 46,XX result in culture and found four (6.1%) of the remaining 65 cases to be consistent with CPM. These cases consisted of normal or mosaic aneuploid cytotrophoblast and non-mosaic aneuploid villous stroma. These studies suggest that each method has specific advantages in the analysis of spontaneous abortions. Direct preparations are less prone to maternal contamination, but certain chromosome abnormalities are more likely to be identified using long-term culture. PMID- 9332659 TI - Prenatal diagnosis of congenital adrenal hyperplasia (21-hydroxylase deficiency) in Croatia. AB - We report on the prenatal diagnosis of congenital adrenal hyperplasia due to 21 hydroxylase in 20 at-risk pregnancies (16 salt-wasting and 4 simple virilizing families). We have diagnosed 3 affected fetuses (2 males and 1 female), 3 healthy homozygotes (2 males and 1 female), and 14 healthy heterozygotes (7 females and 7 males). These data were collected over 4 years. In 16 fetuses, the diagnosis was made with measurements of 17-hydroxyprogesterone (17-OHP) and delta-4 androstenedione (delta) in amniotic fluid (AF), human leukocyte antigen (HLA) typing of amniotic cells, as well as karyotypes between the 16th and 18th weeks of gestation. In 4 fetuses, DNA analysis of amniotic cells was also performed. In 3 pregnancies in which affected fetuses were suspected (on the basis of HLA typing and measurements of 17-OHP and delta concentrations in AF), the fetuses were electively aborted between the 17th to 19th weeks of gestation by parental decision. In all aborted fetuses, diagnosis was confirmed with HLA typing, autopsy findings of hyperplastic adrenal glands, and ambiguous genitalia in female fetuses. Postnatal diagnosis was confirmed in healthy fetuses with HLA typing and serum measurements of 17-OHP concentrations, and in 4 of them with DNA analysis. In 3 of the 4 families, DNA analyses revealed the following mutations: in Family 1, the index case mutation was Intron 2, Exon 3/Exon 6, and the fetus was Normal/Exon 6; in Family 2, the index case mutation was Ex1 Int2 Ex3/ Int2, and the fetus was Ex1 Int2 Ex3/Normal; and in Family 3, the index case mutation was Ex8(356)/Ex8(356), and the fetus was Ex8(356)/ Normal. We also report one case of prenatal diagnosis and treatment. Dexamethasone 0.5 mg BID (20 micrograms/kg/d) was given starting at 6th week of gestation. Prenatal diagnosis suggested, but did not prove, that the female fetus was a heterozygote as the fetus lacked the paternal mutation Ex8(318). No mutation was found in the mother. The fetus, the mother, and the affected sib shared a haplotype, further suggesting heterozygosity. The unaffected status was confirmed postnatally. PMID- 9332660 TI - Tetraphocomelia and bilateral cleft lip in a historical case of Roberts syndrome [Virchow, 1898]. AB - We discuss an unlabelled specimen of tetraphocomelia and bilaterally cleft lip from the former Virchow Museum of our Medical School. Identity of the subject with a case of what was later termed "Roberts syndrome" published by Rudolf Virchow in 1898 is demonstrated. Rediscovery of this important historical case is gratifying, since almost 95% of the specimens of Virchow's collection were lost during World War II. We have restudied Virchow's case. Recent CT scan images of the fetus are presented. We review data from the literature and present new clinical details. The fate of the original clinical data after passing through three reviews is documented briefly. We also reconstruct Virchow's view on phocomelia and its consequences for later research. PMID- 9332661 TI - Bilateral femoral agenesis in femoral facial syndrome in a 19-week-old fetus. AB - Although the cause in most cases is unknown, there is a strong association of the femoral facial syndrome (FFS) with maternal diabetes mellitus. We describe an unusual presentation of FFS in the first pregnancy of a diabetic mother terminated at 19 weeks gestation because of bilateral femoral agenesis diagnosed on ultrasound scan. Autopsy confirmed the absence of the femora and acetabula and the presence of the facial traits of FFS in a female fetus. PMID- 9332662 TI - Ring chromosome 13 with loss of the region D13S317-D13S285: phenotypic overlap with XK syndrome. AB - We report on a patient with a multiple congenital abnormalities/mental retardation (MCA/MR) syndrome including facial abnormalities, agenesis of the corpus callosum, heart defect, 1st ray anomalies of the upper limb, and ambiguous genitalia, whose phenotype overlaps a previous description of XK syndrome. The patient has a ring chromosome (13) with deletion 13q32-qter. Molecular analysis demonstrated loss of the region from D13S317 to D13S285 and a paternal origin of the anomaly. PMID- 9332663 TI - Gene for nonspecific X-linked mental retardation (MRX 47) is located in Xq22.3 q24. AB - We describe a large family with nonspecific X-linked mental retardation (MRX 47). An X-linked recessive transmission is suggested by the inheritance from the mothers in two generations of a moderate to severe form of mental retardation in six males, without any specific clinical findings. Two point linkage analysis demonstrated significant linkage between the disorder and two markers in Xq23 (Zmax = 3.75, theta = 0). Multipoint linkage analyses confirmed the significant linkage with a maximum lod score (Z = 3.96, theta = 0) at DXS1059. Recombination events observed with the flanking markers DXS1105 and DXS8067 delineate a 17 cM interval. This interval overlaps with several loci of XLMR disorders previously localized in Xq23-q24, which are reviewed herein. PMID- 9332664 TI - Multiple congenital anomalies, brain hypomyelination, and ocular albinism in a female with dup(X) (pter-->q24::q21.32-->qter) and random X inactivation. AB - We report on an 18-month-old girl with multiple congenital anomalies (prominence of the metopic suture, fine hair, club foot, absence of the 12th rib, brachydactyly) and severe mental retardation. The funduscopic examination showed diffuse retinal hypopigmentation. Brain magnetic resonance image (MRI) showed signs of diffuse hypomyelination. On cytogenetic and molecular evidence, the karyotype was 46,X,dirdup(X) (pter-->q24::q21.32-->qter). The duplication of the PLP gene, involved in Pelizaeus-Merzbacher disease, was confirmed by fluorescent in situ hybridization (FISH). Both cytogenetic and molecular studies on the X chromosome inactivation status indicated a random pattern in lymphocytes and fibroblasts. This patient appears to be the first case of a female bearing a large duplication of Xq with a random X inactivation. The phenotype of this patient is compared to that of previously reported cases with Xq duplication. PMID- 9332665 TI - Cerebral defects and nephrogenic diabetes insipidus with the ARC syndrome: additional findings or a new syndrome (ARCC-NDI)? AB - We report on 4 children from 2 unrelated families who appear to have the lethal ARC syndrome (arthrogryposis, renal tubular dysfunction, and cholestasis) together with the additional findings of nephrogenic diabetes insipidus and cerebral anomalies, including deafness. With increased survival time in our patients, paucity of the intrahepatic bile ductules and cholestasis progressed to cirrhosis, growth was severely impaired, and severe mental retardation became apparent. No evidence was found for peroxisomal, chromosomal, or mitochondrial disorders. We propose to amend the ARC mnemonic to ARCC-NDI (A-Arthrogryposis, R renal Fanconi, C-cerebral, C-cholestasis, NDI-nephrogenic diabetes insipidus) to name the major manifestations of this syndrome, several of which have not been appreciated. PMID- 9332666 TI - Familial transmission of a small supernumerary marker chromosome 8 identified by FISH: an update. AB - A father and his 2 daughters were previously determined to carry a small, supernumerary marker chromosome [Chudley et al., 1983]. The origin of this marker could not be determined by standard cytogenetic techniques. In this study, fluorescence in situ hybridization (FISH) studies identified the marker chromosome as a pericentric derivative of chromosome 8. The father has low grade mosaicism for this marker and is phenotypically normal. Both daughters are non mosaic and show developmental delays and somewhat differing clinical findings. The phenotypes of the 2 sisters are compared with those previously reported for supernumerary der(8) patients. This is the first report of familial transmission of a supernumerary der(8) marker chromosome. PMID- 9332667 TI - True trisomy 2 mosaicism in amniocytes and newborn liver associated with multiple system abnormalities. AB - Among 58,000 amniocenteses completed, our laboratories found one case of true cytogenetic trisomy 2 mosaicism in a fetus with multiple abnormalities. In contrast, 11 fetuses phenotypically normal at birth were found to have true trisomy 2 mosaicism in their chorionic villus cells among the 10,500 fetuses tested by chorionic villus sampling (CVS). In our single abnormal case, amniocentesis performed at 19 weeks after finding an elevated maternal serum AFP found two independent cultures with trisomy 2 karyotypes in 8 of 25 and 7 of 31 amniocytes, respectively. Although oligohydramnios was noted by ultrasound, the mother elected to continue the pregnancy. At 26 weeks the fetus had intrauterine growth retardation (IUGR), hydronephrosis, and cardiac abnormalities. When delivered by Cesarean section at 30 weeks, the infant had multiple anomalies and developed necrotizing enterocolitis and severe cholestasis. At 5 months coronal magnetic resonance imaging (MRI) displayed delayed myelination and abnormal brain morphology. The patient also exhibited significant growth failure and developmental delay. Although chromosomes were normal in blood, skin fibroblasts, and ascites fluid cells, 4 of 100 hepatic biopsy fibroblasts were 47,XY,+2. Molecular analysis excluded uniparental disomy (UPD) of chromosome 2 in the 46,XY cell line. This and other reports of rare phenotypically abnormal trisomy 2 mosaic fetuses identified by karyotyping amniocytes emphasizes the substantially higher fetal risk of abnormal development than when trisomy 2 is found only in chorionic villus cells. PMID- 9332668 TI - Validation of radiographic criteria for the diagnosis of Down syndrome in stillborn infants. AB - We assessed the utility of radiographic findings as aids to the diagnosis of Down syndrome (DS) in stillborn infants. The iliac index may help to confirm the diagnosis of DS in stillborn infants in whom it is suspected clinically, but in whom it cannot be confirmed cytogenetically. It also can serve as a screening procedure to select stillborns in whom fluorescent in situ hybridization of fixed tissues should be completed. An iliac index of 59 degrees differentiates between control and affected stillborns with the highest accuracy, but false positives persist above 55 degrees, and false negatives are common below 64 degrees. We recommend that a conservative cutoff value of 55 degrees be used if the radiographic data serve as the principal means of diagnosing DS in stillborn infants. A cutoff value of 64 degrees may be appropriate if the radiographic data are used to screen stillborn infants for fluorescent in situ hybridization studies. PMID- 9332669 TI - Direct karyotyping of unstimulated newborn blood: a rapid diagnostic method for the clinical management of critically ill newborns. AB - Using spontaneously dividing nucleated erythrocytes present in newborn cord and peripheral blood, we performed direct karyotype analysis on a cohort of 162 infants suspected of chromosome abnormalities. A cytogenetic diagnosis was obtained in 149 cases (91.9%). In all cases conventional phytohaemagglutinin- (PHA)-stimulated cultures were used for comparison. Concordance between direct and stimulated karyotypes was seen in all but 5 cases. In these 5 cases, abnormalities were seen in the direct harvest while PHA-stimulated cultures showed normal results. Skin fibroblasts from 2 of these cases, available for follow-up, showed the abnormalities in a mosaic state. Our experience confirms that direct karyotyping of fetal and newborn blood is feasible, fast, and efficient and can provide accurate diagnosis of major chromosome abnormalities within 18-24 hours after obtaining the blood. PMID- 9332670 TI - Genomic structure, sequence, and mapping of human FGF8 with no evidence for its role in craniosynostosis/limb defect syndromes. AB - Fibroblast growth factor-8 (Fgf8) is a recently identified growth factor that stimulates the androgen-dependent growth of mouse mammary carcinoma cells. Evidence from mouse development also shows that Fgf8 may play an important role in growth and patterning of limbs, face, and the central nervous system. We describe here the human FGF8 genomic sequence and demonstrate conservation between the human and mouse sequences, including alternatively spliced exons in the mouse. Mapping of FGF8 by FISH using an FGF8-containing bacterial artificial chromosome and by genetic linkage using a SSCP variant identified in this study is also reported and refines the FGF8 map location to 10q24. Since FGF8 maps to the same chromosomal region as FGFR2, has indeed been shown to be a ligand for FGFR2, and has an expression pattern consistent with limb and craniofacial anomalies, we have screened two kindreds with Pfeiffer syndrome that were previously linked to markers from 10q24-25 and a large number of individuals with craniosynostosis and limb anomalies for mutations in the coding sequence of FGF8. While no such mutations were identified, a rare polymorphic variant, consisting of an 18-base-pair (six-amino-acid) duplication in exon 1c, is reported that apparently has no clinical effect. Our exclusionary data suggest that mutations in FGF8 would be, at best, an infrequent cause of such disorders. PMID- 9332671 TI - Limb girdle muscular dystrophy in Manitoba Hutterites does not map to any of the known LGMD loci. AB - Limb girdle muscular dystrophy (LGMD) is a heterogeneous group of disorders affecting primarily the shoulder and pelvic girdles. Autosomal dominant and recessive forms have been identified; 8 have been mapped and 1 more has been postulated on the basis of exclusion of linkage. An autosomal recessive muscular dystrophy was first described in 1976 in the Hutterite Brethren, a North American genetic and religious isolate [Shokeir and Kobrinsky, 1976; Clin Genet 9:197 202]. In this report, we discuss the results of linkage analysis in 4 related Manitoba Hutterite sibships with 21 patients affected with a mild autosomal recessive form of LGMD. Because of the difficulties in assigning a phenotype in some asymptomatic individuals, stringent criteria for the affected phenotype were employed. As a result, 7 asymptomatic relatives with only mildly elevated CK levels were assigned an unknown phenotype to prevent their possible misclassification. Two-point linkage analysis of the disease locus against markers linked to 7 of the known LGMD loci and 3 other candidate genes yielded lod scores of < or = -2 at theta = 0.01 in all cases and in most cases at theta = 0.05. This suggests that there is at least 1 additional locus for LGMD. PMID- 9332672 TI - Another case of the autosomal recessive Weaver-like syndrome. PMID- 9332673 TI - Heterogeneity of X-linked ichthyosis. PMID- 9332674 TI - Conductive hearing loss in the tricho-rhino-phalangeal syndrome (TRP II) or in the Langer-Giedion syndrome. PMID- 9332675 TI - Supra- and infraumbilical raphe with pectus excavatum. PMID- 9332676 TI - SOX3 gene maps near DXS984 in Xq27.1, within candidate regions for several X linked disorders. PMID- 9332677 TI - Heritability of dermatoglyphic a-b ridge count asymmetry. PMID- 9332678 TI - UCNT: chemocurable sometimes, but not very often. PMID- 9332679 TI - Primary extranodal non-Hodgkin's lymphomas. Part 1: Gastrointestinal, cutaneous and genitourinary lymphomas. PMID- 9332680 TI - Randomized study of zorubicin versus zorubicin-cisplatin in undifferentiated carcinoma of the nasopharynx (UCNT) AB - BACKGROUND: The most active chemotherapy regimens in UCNT were those combining anthracyclines (doxorubicin or epirubicin) and cisplatin. Our previous pilot study on 37 patients treated with the zorubicin-cisplatin combination with a RR of 67% and literature data about other anthracyclines such as epirubicin achieving a response rate of over 50% were the basis of this randomized study comparing efficacy and toxicity of the combination vs. zorubicin as monotherapy. PATIENTS AND METHODS: A total of 80 patients entered the study. The diagnosis of UCNT was confirmed by two independent pathologists. All patients had their primary tumors in the nasopharynx. The patients were randomized in two groups: group A (zorubicin 325 mg/m2, day 1), and group B (zorubicin 250 mg/m2, day 1 and cisplatin 30 mg/m2, days 2-5). The inter-cycle interval was four weeks. The two groups were well balanced according to sex, age, stage Ho and TNM stage. RESULTS: Group A: 40 patients included, 34/40 evaluable for activity. Activity on evaluable patient basis: CR 4/34 (11.75%), PR 4/34, SD 14/34, PD 12/34, response rate 8/34 (23.5%); response rate on intent to treat basis 8/40 (20%). TOXICITY: granulocytopenia grade 3-4 6/40, thrombocytopenia grade 3-4 2/40, no febrile neutropenias, nausea/vomiting any grade 3/40, cardiac toxicity any grade (rhythm) 3/40 other toxicities minor or absent. Group B: 40 patients included, 36/40 evaluable for activity. Activity on evaluable patient basis: CR 10/36 (27.78%), PR 17/36, SD 3/36, PD 6/36, response rate 27/36 (75%); response rate on intent to treat basis 27/40 (67.5%). TOXICITY: granulocytopenia grade 3-4 10/40, thrombocytopenia grade 3-4 8/40, two febrile neutropenias, nausea/vomiting any grade 13/40, other toxicities mild or absent. Of the group of patients achieving a CR, four relapsed following 7, 11, 22 and 23 months, one was lost to follow-up, one died after six months from fulminant hepatitis B and eight are in complete remission lasting for 30+ to 66+ months. Following CR achievement none received any consolidation radiotherapy, and the projected five years of freedom from relapse for complete responders is about 60%. CONCLUSION: Zorubicin is an effective drug in UCNT and its combination with cisplatin has a significant activity and an acceptable toxicity. PMID- 9332681 TI - Clinical efficacy and endocrine activity of vorozole in postmenopausal breast cancer patients. Results of a multicentric phase II study. AB - BACKGROUND: Aminoglutethimide was the first aromatase inhibitor to be used successfully in breast cancer patients. However, this drug also inhibits mineralcorticoid and glucocorticoid synthesis, making co-medication with corticosteroids necessary, and it is often poorly tolerated. The primary objective of this trial was to evaluate the clinical efficacy and tolerability of vorozole, a new non-steroidal oral aromatase inhibitor, in postmenopausal breast cancer patients. The secondary objective was to evaluate the pharmacodynamic activity of the drug. SUBJECTS AND METHODS: Thirty-four postmenopausal patients previously treated with tamoxifen in the adjuvant setting and/ or for advanced disease were treated with vorozole, 2.5 mg once daily. Patients were monitored with respect to treatment efficacy and safety. Hormonal evaluations were performed at baseline and during the course of treatment in order to evaluate the pharmacodynamic efficacy and safety of vorozole. RESULTS: According to UICC criteria, there were seven responders, one complete and six partial, for an overall response rate of 21% (95% confidence interval (CI) 9%-38%). The median duration of response was 9.6 months (95% CI 4.6-0), the median time to progression for the entire group was 4.7 months (95% CI 2.9-6.6) and the median survival time was 29.7 months (95% CI 19.1-0). Tolerability was excellent to good in 97% of the patients. Oestradiol and oestrone levels were suppressed to the limit of detection of the assays used. No effect was observed on the other endocrine parameters. CONCLUSIONS: Our results suggest that vorozole is an effective and safe drug for the treatment of advanced postmenopausal breast cancer following tamoxifen failure. PMID- 9332683 TI - The cost-effectiveness of routine postoperative radiotherapy after sector resection and axillary dissection for breast cancer stage I. Results from a randomized trial. AB - BACKGROUND: Cost-effectiveness of routine postoperative radiotherapy after breast conserving surgery has not been prospectively evaluated earlier. In times of rationing of medical resources, valid assessments of cost-effectiveness are important for rational allocation of resources. PURPOSE: Cost and cost effectiveness of routine postoperative radiotherapy was calculated in a prospective randomized trial comparing sector resection plus axillary dissection with (XRT group) or without (non-XRT group) postoperative radiotherapy in breast cancer stage I. Three hundred eighty-one patients were included. After a median follow-up of five years 43 local recurrences, six of them in the XRT-group occurred (P < 0.0001). No difference in regional and distant recurrence (P = 0.23) or survival (P = 0.44) was observed. PATIENTS AND METHODS: Direct medical costs as well as indirect costs in terms of production lost during the treatment period and travel expenses were estimated from data in the medical records and the national insurance registry of each patient. Average costs of different treatment activities and measures were estimated for the XRT-group and the non XRT group respectively. From these estimates differences in costs and effectiveness between the groups were calculated and marginal cost-effectiveness ratios were estimated. For the construction of QALYs each life-year was quality adjusted by a utility value depending on which health state the patient was considered to perceive. RESULTS: Taking into account the cost of primary treatment, the cost of follow-up, the cost of treatment of a local recurrence, travel expenses and indirect costs (production lost) excluding costs for treatment of regional and distant recurrence the cost per avoided local recurrence at five years was SEK 337,727 ($44,438, Pounds 27,018). Adjustment for quality of life showed a cost for every gained QALY to be SEK approximately 1.6 million, ($210,526, Pounds 128,000), range SEK 0.2-3.9 million ($26,315-513,158, Pounds 16,000-312,000). CONCLUSION: The cost of routine postoperative radiotherapy after sector resection and axillary dissection in breast cancer stage I per avoided local recurrence and gained QALY is high. The cost per gained QALY show great variation depending on utility value, which in this study was derived from external observers and not from the patients themselves. These results stress the importance of identifying risk factors for local recurrence, better understanding of impact on quality of life of a local recurrence and adding cost evaluations to clinical trials in early breast cancer. PMID- 9332682 TI - Menstrual cycle and timing of breast surgery in premenopausal node-positive breast cancer: results of the International Breast Cancer Study Group (IBCSG) Trial VI. AB - PURPOSE: It has been postulated that breast cancer surgery performed during the follicular phase of the menstrual cycle is associated with poorer outcome. PATIENTS AND METHODS: We tested this hypothesis by evaluating disease-free survival (DFS) for 1033 premenopausal patients who received definitive surgery either during the follicular phase (n = 358) or the luteal phase (n = 675). All patients were enrolled in a randomized trial conducted between July 1986 and April 1993. All had node positive breast cancer and randomization was stratified by estrogen receptor (ER) status. All patients received at least three cycles of adjuvant cyclophosphamide, methotrexate, and 5-fluorouracil (CMF). The median follow-up was 60 months. RESULTS: Patients who underwent definitive surgery for breast cancer in the follicular phase had a slightly worse disease-free survival than those operated on during the luteal phase (five-year DFS percentage: 53% versus 58%; hazard ratio, 1.13; 95% confidence interval (CI), 0.94-1.38; P = 0.20). The effect was significantly greater for the subpopulation of 300 patients with ER-negative primaries (P = 0.02 interaction effect; five-year DFS percentages 42% vs. 59%; hazard ratio 1.60; 95% CI, 1.12-2.25; P = 0.008). The effect of timing of surgery diminished for analyses based on lesser surgical procedures, e.g., excisional biopsies. In particular, no effect of timing was observed for fine needle aspiration procedures. CONCLUSIONS: Surgical procedures which are more extensive than a fine needle aspiration biopsy might be associated with worse prognosis if conducted during the follicular phase of the menstrual cycle. This phenomenon was seen predominantly for high risk breast cancer with low levels or no estrogen receptors in the primary tumor. PMID- 9332684 TI - Long-term follow-up and post-relapse survival in patients with non-metastatic osteosarcoma of the extremity treated with neoadjuvant chemotherapy. AB - BACKGROUND: Most of the studies of the treatment of non-metastatic osteosarcoma of the extremity have reported results in terms of probability of survival up to five years with a minimum follow-up of less than two to three years. Definition of reliable indicators of prognosis and predictive factors for survival require mature data derived from a long-term survival analysis. PATIENTS AND METHODS: A review of 127 patients with non-metastatic osteosarcoma of the extremity, treated between March 1983 and June 1986, was performed. The treatment protocol consisted of primary chemotherapy with MTX (randomization to high vs. moderate dosages) and CDP followed by surgery. Postoperatively, patients with < 60% tumor necrosis received ADM and BCD; those with tumor necrosis > or = 60% < 90% (Fair Responders FR) were given MTX, CDP and ADM. Up to January 1984, patients with tumor necrosis > 90% received MTX and CDP only, and after then they were given the same treatment as for FR. A multivariate analysis to test predictive factors for survival was performed. RESULTS: With a median follow-up of 134 months (range 114 153), the 12-year DFS was 46%. A good histologic response, an LDH baseline value within the normal range, and the use of high-dose MTX were positive predictive factors for DFS. With a median time of observation for survivors of 130 months, the 12-year overall survival was 53%. None of the patients who relapsed with local or distant recurrences other than lung metastasis are now alive. Patients with a relapse-free interval longer than 24 months had a significantly better post-relapse survival than those with a shorter relapse-free interval (40% vs. 7%; P = 0.0159). All of the patients who were not surgically treated had disease progression and died within 40 months after the first recurrence. The surgically treated patients had a 30% post-relapse survival probability. CONCLUSIONS: In non metastatic osteosarcoma of the extremity, chemotherapy-induced tumor necrosis, the baseline LDH serum value and the use of HDMTX are significant predictive factors for DFS. The relapse-free interval and the possibility of metastasectomy are significant factors conditioning the post-relapse survival. PMID- 9332685 TI - Treatment with cisplatin and fluorouracil alternating with radiation favourably affects prognosis of inoperable squamous cell carcinoma of the head and neck: results of a multivariate analysis on 273 patients. AB - PURPOSE: The goal of the present analyses is to assess the association between different therapeutic approaches and both the probability of achieving a complete response and the risk of death in patients with stage III-IV, inoperable, squamous cell carcinoma of the head and neck (SCC-HN). PATIENTS AND METHODS: Between August 1983 and December 1990, 273 patients with stage III-IV, previously untreated, unresectable SCC of the oral cavity, pharynx and larynx, were included into two consecutive randomized multi-institutional trials (HN-7 and HN-8 protocols) coordinated by the National Institute for Cancer Research (NICR) of Genoa. The HN-7 protocol compared neo-adjuvant chemotherapy (four cycles of vinblastine, 6 mg/m2 i.v. followed by bleomycin, 30 IU i.m. six hours later, day 1; methotrexate, 200 mg i.v., day 2; leucovorin, 45 mg orally, day 3) (VBM) followed by standard radiotherapy (70-75 Gy in 7-8 weeks) (55 patients) to alternating chemoradiotherapy based on four cycles of the same chemotherapy alternated with three splits of radiation, 20 Gy each (61 patients). In the HN-8 protocol standard radiotherapy (77 patients) was compared to the same alternating program as the one used in the previous protocol but employing cisplatin, 20 mg/m2/day and fluorouracil, 200 mg/m2/day, bolus, both given for five consecutive days (CF) instead of VBM (80 patients). A single database was created with the patients on the two protocols. Age at diagnosis, gender, site of the primary tumor, size of the primary, nodal involvement, performance status and treatment approach were analyzed by the multiple logistic regression model and the Cox regression method. The analyses were repeated including the treating institutions as a covariate (coordinating center versus others). RESULTS: The multiple logistic regression analysis indicates that treatment (alternating more so than others, regardless of the chemotherapy regimen used) (P = 0.0001) is more likely to be associated with complete response. In addition, size of the primary tumor (P = 0.004), nodal involvement (P = 0.02) and performance status (P = 0.009) are prognostic variables affecting the probability of achieving a complete response. The Cox regression analysis indicates that treatment, performance status, size of the primary tumor, nodal involvement and, marginally, site of the primary tumor, are independent prognostic variables affecting the risk of death. When the radiation-alone therapy is adopted as the reference treatment, the relative risk of death is 0.58 (95% confidence interval (CI) 0.40-0.84) for alternating CF and radiation, 0.79 (95% CI 0.53-1.16) for alternating VBM and radiation and 1.30 (95% CI 0.89-1.92) for sequential VBM and radiation. When the treating institution is included in the model, a 34% increased risk of death (P = 0.04) is observed for patients treated outside the coordinating center. CONCLUSION: In our series of patients with advanced, unresectable SCC-HN, treatment with cisplatin and fluorouracil alternating with radiation was associated with a more favourable prognosis. The role of the treating institution in the modulation of the treatment outcomes was also relevant. PMID- 9332686 TI - Prognostic impact of an activation of coagulation in lung cancer. AB - BACKGROUND: There is evidence that activation of coagulation by influencing tumour biology may have impact on clinical course of lung cancer. PATIENTS AND METHODS: We measured the activation markers thrombin-antithrombin complex (TAT) and prothrombin fragment F1 + 2 in 99 lung cancer patients immediately after diagnosis, before antineoplastic treatment. Outcome was assessed at the end of appropriate standard primary therapy (four to six courses of chemotherapy, surgery or radiation). RESULTS AND CONCLUSIONS: The activation markers (means +/- SEM) were lower in the 33 responders (RSP; complete or partial remission) than in the 66 non-responders (NRSP): TAT 3.96 +/- 0.48 vs. 9.69 +/- 1.57 micrograms/l (P < 0.001), and F1 + 2 1.09 +/- 0.09 vs. 1.64 +/- 0.25 nmol/l (P < 0.05). TAT levels were > 6 micrograms/l in 30 of 66 (45%) NRSP, but only 4 of 33 (12%) RSP. 88% of patients with TAT < or = 6 micrograms/l achieved remission, and 45% with TAT > 6 micrograms/l (P = 0.0014). In the subgroup of 46 patients with advanced disease, the six RSP showed lower TAT than the 40 NRSP: 4.65 +/- 0.94 vs. 11.92 +/- 2.49 micrograms/l (P < 0.01); one of six (17%) RSP, but 21 of 40 (53%) NRSP showed TAT > 6 micrograms/l. These data suggest that in lung cancer the activation of coagulation is an independent prognostic factor, since TAT levels were different between RSP and NRSP, also within the homogeneously unfavourable metastatic subgroup. It should be further studied, whether TAT can identify patients, whose prognosis could be improved by anticoagulation as an adjunct to standard antineoplastic therapy. PMID- 9332688 TI - Solitary meningeal plasmacytomas. AB - BACKGROUND: Solitary extramedullary plasmacytoma (SEP) represents a rare separate clinical pathological entity; it is radiosensitive, curable, and unrelated to myeloma. Only 15 cases have been reported. PATIENTS AND METHODS: Authors report two well documented cases of dural plasmocytomas mimicking meningiomas. This paper points out differences between plasmacytomas, with very different initial presentation, and other meningeal tumors including extensive radiological investigations. Diagnostic options and outcomes are discussed. CONCLUSION: With solitary plasmacytomas, radiological diagnosis is difficult to assess. Serum is generally free of monoclonal protein. The importance of the most recent techniques in ruling out other pathological entities is stressed. Histopathological examination is required to document a monoclonal type of plasma cell tumor. Treatment includes surgery and radiotherapy. Differential radiological diagnosis will be discussed. PMID- 9332689 TI - Treatment of advanced pancreatic cancer with epirubicin, 5-fluorouracil and 1 leucovorin: a phase II study. AB - BACKGROUND: Preclinical data suggest that the levorotatory isomer of leucovorin (1-LV) can augment the cytotoxic effects of 5-fluorouracil (5-FU) more effectively than racemic LV. A phase II study was initiated to evaluate the effect of a combination of the pure L-isomer of LV and 5-FU along with epirubicin in patients with advanced pancreatic cancer. PATIENTS AND METHODS: Twenty-eight consecutive, previously untreated patients with measurable metastatic adenocarcinoma of the pancreas were enrolled in this study. Treatment consisted of epirubicin 50 mg/m2 on day 1 and 5-FU 400 mg/m2 plus 1-LV 100 mg/m2 both administered on days 1 to 5. Treatment cycles were repeated every four weeks for a total of six months unless progressive disease was documented earlier. The study endpoints were survival, toxicity, objective response as well as palliative beneficial effects of the regimen in terms of performance status, body weight and pain. RESULTS: Six patients had a partial response (21%; 95% CI, 8% to 44%) and 10 (36%) stable disease (35%), while the remaining 12 patients (43%) progressed during treatment. The median time to treatment failure was 2.9 months (range 1.2 13.7 months), and the median survival time was six months (range 1.8-19 months), with three patients (11%) being alive after one year. Beneficial palliative effects in terms of performance status, body weight and/or pain were seen in 12 of 28 patients (43%): an improvement in performance status was recorded in four patients, pain was attenuated and/or analgetic consumption reduced by > or = 50% in eight patients, and five patients had weight gain of > 5% of their pretreatment body weight. Treatment-associated side effects were generally mild to moderate. Severe adverse reactions included (asymptomatic) granulocytopenia in five, mucositis and/or diarrhea WHO grade 3 in four patients. CONCLUSION: Our data suggest that epirubicin plus 5-FU and 1-LV is a tolerable regimen that has some positive effects in the treatment of pancreatic cancer. Since this regimen, however, is unlikely to offer any major therapeutic advantage compared to monochemotherapy or other combination regimens, the search for novel and more effective cytotoxic agents must continue. PMID- 9332687 TI - Index of pretreatment intensity predicts outcome of high-dose chemotherapy and autologous progenitor cell transplantation in chemosensitive relapse of Hodgkin's disease. AB - PURPOSE: To identify prognostic factors in patients with chemosensitive relapsed Hodgkin's disease treated by high-dose chemotherapy with autologous progenitor cell transplantation (HDC) and to compare the duration of treatment-free remission prior to HDC with the progression-free survival after HDC in individual patients. PATIENTS AND METHODS: Forty-five consecutive patients were analyzed retrospectively. We devised an index of pretreatment intensity (IPTI) based number of different chemo- and radiotherapy regimens given between diagnosis and HDC and on the duration of disease. RESULTS: With a median follow-up of 47 months the post-transplant event-free survival (EFS) was 44% and the overall survival (OAS) was 62% at four years. The IPTI allowed to discriminate between a low and a high-risk group with a four-year post-transplant EFS of 66% and 11% and a OAS of 87% and 28%, respectively (P = 0.0001). Of the 39 patients with sufficient follow up after HDC, post-transplant EFS lasted on average > or = 18.5 months longer than the pretransplant treatment-free remission. CONCLUSIONS: HDC with the CBV regimen confers significant benefit to patients with chemosensitive relapsed Hodgkin's disease. The IPTI may help to select patients with a good response to HDC and to identify poor prognosis patients suitable for experimental protocols or palliative care only. PMID- 9332690 TI - Successful intraperitoneal suramin treatment of peritoneal mesothelioma. PMID- 9332691 TI - Paclitaxel in untreated FIGO stage III suboptimally resected ovarian cancer. AB - BACKGROUND: We wanted to assess the efficacy and toxicity of paclitaxel (Taxol) in previously untreated patients with advanced ovarian cancer. No such study had been performed at the time of initiation of this study. PATIENTS AND METHODS: The study population in this analysis consisted of 35 previously untreated stage III ovarian cancer patients, suboptimally resected at primary surgery. The initial paclitaxel dose was 175 mg/m2 given as a three-hour i.v. infusion every three weeks. RESULTS: A total of 225 paclitaxel courses were given to 35 patients of whom 34 were evaluable for clinical response. Nine (26%) patients obtained a complete response to paclitaxel, ten (29%) a partial response, seven (21%) stable disease and eight (24%) had progressive disease. Thus, the total response rate to paclitaxel treatment was 55% (19 of 34). The median duration of response for the 19 responding patients was eight months (range 1-44.5+). The median duration for nine patients with clinical complete response was 16 months (range 2-44.5+). A second-look laparotomy was performed in 15 patients after six courses of paclitaxel. Of these, five obtained a histopathologically complete remission, five a partial remission and five stable disease. The five patients with pathologically complete remissions are alive with a median progressive-free survival of 24.5 months (range 15(+)-44.5+). The median progression-free survival for all patients was 6.1 months (range 1-44.5+). Toxicity consisted mainly of neutropenia, easily controlled. Other toxicities were myalgia/arthralgia and peripheral neuropathy. Three patients experienced a severe anaphylactic reaction during the first course. CONCLUSION: This study showed that paclitaxel is an effective and safe drug for first-line treatment of ovarian cancer. PMID- 9332693 TI - Clinical manifestation of severe hyperammonaemia in patients with multiple myeloma. PMID- 9332692 TI - Phase I study of 3'-deamino-3'-(2-methoxy-4-morpholinyl)doxorubicin (FCE 23762, PNU 152243) administered on a daily x3 schedule. AB - BACKGROUND: 3'-Deamino-3'-(2-methoxy-4-morpholinyl)doxorubicin (FCE 23762, PNU 152243) is a highly lipophilic doxorubicin derivative which possesses potent in vitro and in vivo antitumor activity. Previous phase I studies had been conducted using a single bolus every 28 days. PATIENTS AND METHODS: We conducted a phase I study of FCE 23762 on a daily x3 every 28 days schedule. Thirty patients received 68 cycles of therapy at 5 dose levels (200-600 micrograms/m2/d). RESULTS: Prolonged neutropenia and thrombocytopenia were the dose-limiting toxicities. Other nonhematological toxicities included nausea and vomiting, anorexia, fatigue and transient elevations of serum creatinine and hepatic transaminases. No cardiac toxicity was demonstrated. There were no partial or complete antitumor responses. CONCLUSION: The recommended phase II dose using the schedule defined in this study is 500 micrograms/m2/dx3. PMID- 9332694 TI - Megestrol activity in recurrent adult type granulosa cell tumour of the ovary. PMID- 9332695 TI - Ataxia following docetaxel infusion. PMID- 9332696 TI - Control of the mutagenicity of aromatic amines by protein kinases and phosphatases. I. The protein phosphatase inhibitors okadaic acid and ortho vanadate drastically reduce the mutagenicity of aromatic amines. AB - The role of protein kinase C and protein phosphatases was examined in the control of mutagenic metabolites of aromatic amines. Various metabolic activating systems derived from rat liver were treated with: 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C modulator; okadaic acid (OA), a potent inhibitor of serine/threonine protein phosphatases (PP1 and PP2A); and ortho-vanadate (OV), an inhibitor of tyrosine phosphatases. TPA used over a wide concentration range (10( 9)-10(-6) M) did not affect the bacterial mutagenicity of the aromatic amines and of the aromatic amide investigated, 2-aminoanthracene, 2-aminofluorene and 2 acetylaminofluorene (2AAF). At the molecular level, TPA did not affect the function of cytochrome P450s 1A1 or 1A2, which are known key factors for the activation and inactivation of aromatic amines/amides. By contrast the OA and OV treatment of rat hepatocytes, rat liver homogenate, fraction S9 and the nuclear fraction drastically reduced (by > 80%) the mutagenicity of the aromatic amines/amide investigated. This is by far the most pronounced change in genotoxicity observed to date via modulation of phosphorylation. Whilst the mutagenicity of the primary toxication product 2-N-OH-acetylaminofluorene (2-N-OH AAF) in the presence of exogenous activating systems (hepatocytes, S9-fraction, nuclear fraction) was also reduced by OV, OA had no influence. Thus the tyrosine protein phosphatase inhibitor and the serine/threonine protein phosphatase inhibitor influence the genotoxicity of aromatic amines/amides on different levels. Moreover, this shows that the drastic reduction in mutagenicity by OA was due to its influence on a step prior to the presence of the primary toxication product 2-N-OH-AAF. This reduction could be due to changes in the activity of cytochrome P4501A1 and/or 1A2. However, no incorporation of 32P-labelled phosphate from intracellularly prelabelled [32P]-ATP into cytochromes P450 1A1 or 1A2 nor any change in their catalytic activities was observed in the presence of OA. Furthermore, a phosphorylation dependent change in the function of P glycoprotein (known for its role in the transport of diverse xenobiotic substances and their metabolites) was shown not to contribute to the observed decrease in mutagenicity. Our results reveal an important role for protein phosphatase 1 and/or 2A and tyrosine phosphatase(s) in the control of the genotoxicity of aromatic amines and amides. However, the present study does not distinguish between effects mediated by individual proteins affected by these protein phosphatases. PMID- 9332697 TI - Cisplatin-induced apoptosis of immortalized mouse proximal tubule cells is mediated by interleukin-1 beta converting enzyme (ICE) family of proteases but inhibited by overexpression of Bcl-2. AB - Cisplatin is known to induce serious renal damage including acute renal failure, the major site of renal injury appears to be localized to the third segment of the proximal tubule (S3). Apoptosis occurs during a variety of acute injuries to tubule cell. The purpose of this study was to determine whether cisplatin induces apoptosis of immortalized mouse S3 cells, and to define the intracellular pathways leading to cell death. S3 cells exposed to cisplatin exhibited biochemical, morphological, and flow cytometric changes characteristic of apoptosis associated with slight necrosis. Cisplatin-induced apoptosis could be inhibited by overexpression of crmA, a cowpox virus gene, of which the product is known to suppress activities of the interleukin-1 beta converting enzyme (ICE) family proteases. On the other hand, overexpression of bcl-2, an antiapoptotic oncogene, rendered S3 cells partially resistant to cisplatin. These results indicate that cisplatin-induced proximal tubule damage is associated with apoptosis, which is positively modulated by the ICE family of proteases and negatively by the product of bcl-2. PMID- 9332698 TI - Relationship of anesthetic activity of alkyl acetates to hydrophobicity and in vivo effect on membrane fluidity in mice. AB - In vivo anesthetic activity of alkyl acetates in mice was studied in relation to hydrophobicity and the in vivo effect on membrane fluidity. The anesthetic potency (AD50) of alkyl acetates was determined; AD50 shows the i.p. dose required to anesthetize 50% of mice from the treated group. We used log P (n octanol/water partition coefficient) as an operational definition of hydrophobicity. Membrane fluidity was determined using 1,6-diphenyl-1,3,5 hexatriene (DPH) as fluorescence probe. Log (1/AD50) was a parabolic function of log P, and the value of log P that corresponds to the minimum AD50 was estimated to be 2.08. Brain synaptosomal membranes were prepared from mice 30 min after dosing with each of the three alkyl acetates applied at 1.5-fold AD50: n-butyl, n amyl, and n-hexyl acetate. In each alkyl acetate group, most of the animals were anesthetized (> 68%). Decreased membrane fluidity was observed for the animals that were anesthetized while no change in the fluidity was seen for the animals that were not anesthetized. The results suggest an involvement of decreased DPH fluidity in alkyl acetate-induced anesthesia. PMID- 9332699 TI - Effect of linoleic acid, linoleic acid anilide, and arachidonic acid on the expression of adhesion molecules on human neutrophils. AB - The effects of linoleic acid, linoleic acid anilide, and arachidonic acid on the expression of CD11b/ CD18, CD11c/CD18 integrins and L-selectin on human neutrophils were studied by flow cytometry in a whole blood assay. None of these compounds had any effect on the basal expression of CD11b, CD11c, or L-selectin in the concentration range of 20-100 microM. However, linoleic acid at a concentration of 1000 microM slightly up-regulated CD11b and CD11c by a factor of 2.1 and 1.7, respectively. Linoleic acid, linoleic acid anilide, and arachidonic acid did not affect the formyl-methionyl-leucyl-phenylalanine induced up regulation of CD11b or CD11c. However, linoleic acid and linoleic acid anilide slightly inhibited the phorbol myristate acetate (PMA)-induced expression of CD11b, which was decreased by 27 and 21% at concentrations of 100 and 1000 microM, respectively. Likewise, arachidonic acid at 40 microM inhibited the PMA induced expression of CD11b by 19%. Our results suggest that linoleic acid, linoleic acid anilide, and arachidonic acid do not dramatically affect the expression of leukocyte adhesion molecules in a whole blood assay. PMID- 9332700 TI - Inhibitory effect of nicotinamide to paraquat toxicity and the reaction site on complex I. AB - The inhibitory effect and mechanism of action of nicotinamide to paraquat toxicity were studied in male Sprague-Dawley rats. Proteins of submitochondrial particles (SMP), especially of mol. wt. 25-30 kDa, in rat lungs were destroyed by paraquat radicals, and aggregated protein bands approximately 100 kDa were observed by polyacrylamide electrophoresis. The competitive inhibition effects were observed of nicotinamide on NADH oxidation by paraquat via SMP in rat lungs and the Ki was 9.3 mM. The inhibitory effects of nicotinamide on lipid peroxidation by paraquat with rat lung and liver SMP were verified. The times of occurrence of dyspnea and death in rats after paraquat exposure were delayed by nicotinamide administration. The activity of NADH: ubiquinone reaction of NADH:ubiquinone oxidoreductase (complex I) in rat lung was reduced 24 h after paraquat exposure, and was protected by nicotinamide. The activity of NADH:ferricyanide reaction of complex I was, however, reduced by administration not only of paraquat but also nicotinamide. These results imply that nicotinamide is inhibitory to paraquat toxicity. Nicotinamide, paraquat, and ferricyanide may react at overlapping sites on complex I. PMID- 9332701 TI - Time courses of hepatic injuries induced by chloroform and by carbon tetrachloride: comparison of biochemical and histopathological changes. AB - The relationship was investigated between biochemical and morphological changes in chloroform (CHCl3)- and carbon tetrachloride (CCl4)-induced liver damage. The time courses of hepatic microsomal cytochrome P450 (CYP) content, hepatic microsomal CYP2E1 activity, hepatic reduced glutathione (GSH) content, plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were examined in relation to the liver morphology in rats orally treated with CHCl3 or CCl4 (3.35 mmol/kg). The CYP content and the activity of CYP2E1 markedly decreased in the CCl4-treated rats 3 h after treatment compared to much lower decreases in the CHCl3-treated rats. The hepatic GSH content was decreased to a similar extent in both groups of rats at 3 h after treatment; in the CCl4-treated rats, the GSH content continued to decrease, reaching a minimum at 24 h and without attaining the normal level at 72 h after treatment. By contrast, hepatic GSH content in the CHCl3-treated rats began to increase from 6 h, attaining complete recovery 48 h after treatment. Plasma ALT and AST activities were significantly elevated by CCl4 as early as 3 h after treatment, while the activities in the CHCl3-treated rats did not increase until 6 h after treatment. In both groups of rats, ALT and AST activities reached a maximum at 24 h, and gradually decreased, remaining at abnormal levels at 72 h. Hepatic cells in the CCl4-treated rats were found to be necrotic as early as 3 h post-treatment, whereas few or no morphological changes appeared in the liver of CHCl3-treated rats. The extent of necrosis was at a maximum 24 h after treatment in both CHCl3- and CCl4-treated rats. In addition, some necrotic cells remained in the liver of CCl4-treated rats 72 h after treatment, while the necrosis in the CHCl3-treated rats was almost negligible. The present results indicate that almost the same time-courses of biochemical and morphological changes were followed in rats of both the CHCl3- and CCl4-treated groups. PMID- 9332702 TI - Effect of phenobarbital on hepatic eicosanoid concentrations in rats. AB - Phenobarbital is an efficacious tumor-promoting agent in the liver. Studies using inhibitors of eicosanoid synthesis have suggested that eicosanoids are important in the promotion of hepatocarcinogenesis by phenobarbital. We therefore hypothesized that hepatic eicosanoid concentrations are altered following phenobarbital administration. Male Sprague-Dawley rats were fed one of four levels of phenobarbital (0, 0.02, 0.05, and 0.1%). Eight rats from each of the four groups were killed after 10, 24, and 44 days for determination of liver weight and for preparation of microsomes. No significant difference was found among rat weights; however, liver weights were significantly higher in rats fed phenobarbital. Assay of benzyloxyresorufin-O-dealkylase activity showed that cytochromes P-450 2B1 and 2B2 were induced in response to phenobarbital administration. Prostaglandin E2 concentrations were found to be significantly decreased by phenobarbital treatment after 10 and 24 days, but not after 44 days. Prostaglandin F2 alpha levels were decreased only by the lowest dietary phenobarbital concentration. Hepatic concentrations of leukotriene C4 were decreased significantly at 10 days and at 44 days (only for the group administered the highest percentage concentration of phenobarbital), but not at 24 days. These results show that the investigated eicosanoids are generally slightly decreased by phenobarbital administration. Elevated eicosanoid levels therefore do not appear to be necessary for the promoting activity of phenobarbital. PMID- 9332703 TI - Pyruvate alleviates toxic effects of ethanol on cells in culture. AB - Cytotoxic effects of ethanol on cultured human hepatocytes and fibroblasts differed with the type of culture medium used. When cytotoxic effects of ethanol were compared between culture systems using either RPMI-1640 or Dulbecco's modified Eagle's medium (DMEM), the cytotoxicity was more prominent with the former medium than with the latter. A reduction of the cytotoxic effects appeared to be due to the pyruvate contained in DMEM. The protective effect of pyruvate against ethanol toxicity may be related to tricarboxylic acid (TCA) cycle activity because addition of malate to the medium also reduced the cytotoxic effects. Our results suggest that drug cytotoxicity testing in vitro must be done using various types of culture medium. PMID- 9332705 TI - Decision support in the operating theatre and intensive care: a personal view. PMID- 9332704 TI - Control of the mutagenicity of arylamines by protein kinases and phosphatases: II. Lack of response of rat liver N-acetyl transferases to phosphorylation modulators. AB - Treatment of rat hepatocytes with the phosphatase inhibitors okadaic acid or ortho-vanadate had led to an 80% decrease in the bacterial mutagenicity of several aromatic amines metabolically activated by these hepatocytes. This is the most dramatic change yet demonstrated in mutagenicity by phosphorylation modulation. However, incorporation of phosphate into and catalytic activity of cytochromes P450 (CYP) 1A1 and 1A2, the major catalysts for the first step in the toxication of aromatic amines, were unchanged. We therefore investigated whether changes in the phosphorylation status would influence the activities of the N acetyltransferases NAT1 and/or NAT2, being responsible for one of the two major pathways leading to the ultimate mutagens, the reactive esters which are derived from the N-hydroxylated metabolites of aromatic amines. Hepatocytes were derived from the livers of rats pretreated with CYP1A1/1A2 inducers and from untreated rats using conditions under which the phosphorylation-dependent drastic decrease of the arylamine mutagenicity was observed. Treatments were exposure to 1 mM dibutyryl-cAMP (protein kinase A stimulator), 100 nM okadaic acid or 20 nM calyculin A (preferential inhibitors of serine/threonine phosphatases PP2A and PP1, respectively), 2 mM ortho-vanadate (inhibitor of tyrosine phosphatases), and 50 mM NaF (stimulator of adenylate cyclase and non-specific inhibitor of protein phosphatases). None of the phosphorylation modulators led to a significant change in NAT1 or NAT2 activities. This was true for hepatocytes from rats which had been pretreated with inducers for CYP1A1 and CYP1A2 as well as from untreated rats. The inducers led to the expected increases in CYP1A1 and CYP1A2 but the NAT1 and NAT2 activities remained unchanged. Our study shows that the N-acetyl transferases NAT1 or NAT2, the catalysts responsible for the formation of the highly reactive N-acetoxy derivatives of N-hydroxylated aromatic amines, are not responsible for the drastic decrease in arylamine genotoxicity after treatment of the metabolizing system with protein phosphatase inhibitors. The data also show that NAT1 and NAT2 are not regulated by the classical xenobiotic metabolizing enzyme inducers nor by any of the phosphorylation modulators used. PMID- 9332706 TI - NeoGanesh: a working system for the automated control of assisted ventilation in ICUs. AB - Automating the control of therapy administered to a patient requires systems which integrate the knowledge of experienced physicians. This paper describes NeoGanesh, a knowledge-based system which controls, in closed-loop, the mechanical assistance provided to patients hospitalized in intensive care units. We report on how new advances in knowledge representation techniques have been used to model medical expertise. The clinical evaluation shows that such a system relieves the medical staff of routine tasks, improves patient care, and efficiently supports medical decisions regarding weaning. To be able to work in closed-loop and to be tested in real medical situations, NeoGanesh deals with a voluntarily limited problem. However, embedded in a powerful distributed environment, it is intended to support future extensions and refinements and to support reuse of knowledge bases. PMID- 9332707 TI - Evaluation of a medical diagnosis system using simulator test scenarios. AB - This paper describes an informal but systematic method for how to test and verify a knowledge-based system in a large open-ended medical target domain. The system used is Guardian, an intelligent system for monitoring and diagnosis of post cardiac surgery patients in an intensive-care unit. The knowledge base is tested and verified by running the system on a series of realistic test scenarios, both with an embedded simulator and with an external simulation system. The same scenarios are presented to human test subjects, making it possible to compare and analyze the performance of the knowledge-based system with that of human physicians. The use of simulators instead of clinical data also means that it is possible to test crucial scenarios which occur seldom in medical practice. Our results show that a system like Guardian might indeed be useful in medical care. PMID- 9332708 TI - A decision-driven design of a decision support system in anesthesia. AB - We present a new approach to the design of a decision support system (DSS) in anesthesia which converts the available data to relevant information. Instead of a patient-driven design (patient modelling), we use a decision-driven design (anesthetist modelling). This approach results in a system consisting of three stages. First, the incoming data is validated to ensure reliable further processing for both DSS and anesthetist. Second, the validated data is analyzed to detect patterns that could trigger the anesthetist's response. Finally, from these patterns a strategic selection is made and offered to the anesthetist as information relevant for the decision that has to be made in the current context. Results show that this approach is feasible, but further evaluation is necessary before practical application is possible. PMID- 9332709 TI - Recognition of patient anaesthetic levels: neural network systems, principal components analysis, and canonical discriminant variates. AB - The goal of this study was to examine the ability of Neural Networks to recognise the levels of anaesthetic state of a patient. Data obtained under different levels of anaesthesia have been modelled for the purpose. It is shown that inferential parameters can be used to recognise the levels of anaesthesia. In addition to demonstrating the ability of neural networks for classification we were interested in understanding the classification strategy discovered by the neural networks. Multivariate data analysis techniques, namely Principal Components Analysis and Canonical Discriminant Variates, were applied to analyse the resultant networks. PMID- 9332710 TI - Malate dehydrogenase isozymes (MDH; EC 1.1.1.37) in long-term callus culture of Cereus peruvianus (Cactaceae) exposed to sugar and temperature stress. AB - Malate dehydrogenase (MDH; EC 1.1.1.37) isozymes in long-term callus tissue culture of Cereus peruvianus were studied in starch gel electrophoresis to investigate the control of differential Mdh gene expression under sugar and temperature stress. While two cytosol MDH isozymes showed an unchanged phenotype when the callus tissues were transferred to medium maintained at 22 or 37 degrees C and containing different concentrations of sucrose, glucose, and fructose, the different combinations of five mitochondrial MDH (mtMDH) and two micro-body MDH (mbMDH) showed different MDH isozyme patterns in the callus populations. Differential expression of mtMDH isozymes seems to be modulated at the posttranslational level in callus tissues exposed to different concentrations and types of sugar and to high-temperature and low-temperature stress. An inductor effect on the expression of mbMDH isozymes was observed under stress conditions and in long-term callus tissue, and they may also present different responses. PMID- 9332711 TI - Proboscidean DNA from museum and fossil specimens: an assessment of ancient DNA extraction and amplification techniques. AB - Applications of reliable DNA extraction and amplification techniques to postmortem samples are critical to ancient DNA research. Commonly used methods for isolating DNA from ancient material were tested and compared using both soft tissue and bones from fossil and contemporary museum proboscideans. DNAs isolated using three principal methods served as templates in subsequent PCR amplifications, and the PCR products were directly sequenced. Authentication of the ancient origin of obtained nucleotide sequences was established by demonstrating reproducibility under a blind testing system and by phylogenetic analysis. Our results indicate that ancient samples may respond differently to extraction buffers or purification procedures, and no single method was universally successful. A CTAB buffer method, modified from plant DNA extraction protocols, was found to have the highest success rate. Nested PCR was shown to be a reliable approach to amplify ancient DNA templates that failed in primary amplification. PMID- 9332712 TI - A unique cDNA coding for subunits 8 and 6 of mitochondrial adenosine triphosphatase of the lancelet Branchiostoma lanceolatum, an ancestor of vertebrates. AB - A cDNA encoding subunits 8 and 6 of mitochondrial ATPase of the cephalochordate lancelet (Branchiostoma lanceolatum), a direct ancestor of vertebrates, has been cloned and sequenced. A unique transcript encodes the 8 and 6 subunits. In the course of isolation of the 5' end of the ATPase 8 subunit gene, we also determined the sequence of the 5' adjacent tRNA-Lys gene. The anticodon used by mitochondrial tRNA-Lys is TTT. PMID- 9332713 TI - Isozyme variability in plants regenerated from calli of Cereus peruvianus (Cactaceae). AB - Morphological and isozyme variation was observed among plants regenerated from callus cultures of Cereus peruvianus. Different morphological types of shoots (68%) were observed in 4-year-old regenerated plants, while no distinct morphological variants were observed in plants grown from germinated seeds. Isozyme patterns of 633 plants regenerated from calli and of 261 plants grown from germinated seeds showed no variation in isocitrate dehydrogenase isozyme, and the differential sorbitol dehydrogenase, alcohol dehydrogenase, malate dehydrogenase, acid phosphatase, and peroxidase isozyme patterns observed in regenerated plants were attributed to nonallelic variation. Allelic variation was detected at three isoesterase loci. The proportion of polymorphic loci for both populations was 13.6% and the deviation from Hardy-Weinberg equilibrium for the Est-1 and Est-7 loci observed in somaclones was attributed to the manner in which the regenerant population was established. The high values for genetic identity among regenerant and seed-grown plant populations are in accordance with the low levels of interpopulation genetic divergence. In somaclones of C. peruvianus, morphological divergence was achieved within a short time but was not associated with any isozyme changes and also was not accompanied by biochemical genetic divergence. PMID- 9332714 TI - Isozyme extraction from shoot tissue of Cereus peruvianus (Cactaceae) for electrophoretic analysis. PMID- 9332715 TI - Global cerebral ischemia activates nuclear factor-kappa B prior to evidence of DNA fragmentation. AB - The oxidative stress responsive transcription factor nuclear factor-kappa B (NF kappa B) consists of a p50 (50 kDa) and p65/RelA (65 kDa) component and can be activated in vitro by TNF alpha, IL1 beta, hydrogen peroxide and oxygen radicals. All of the above factors are also known to be elevated at certain times after transient global ischemia. The present study was performed to determine if NF kappa B was activated in vivo by transient global forebrain ischemia. Adult male rats were subjected to 30 min of 4-vessel occlusion (4-VO) and sacrificed at selected post-ischemic time points. Levels of NF-kappa B p50 and p65 subunits were determined by immunocytochemistry, Western blot and electrophoretic mobility shift analysis. The enhancer complex was also confirmed by immuno-gel-shift analysis. Specific labeling of DNA strand breaks and DNA fragmentation was examined in situ by means of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. Western blot analysis of hippocampus showed induction of p50 and p65. A time course of NF-kappa B induction in hippocampus showed a p50-specific band at 6 h that increased in intensity over 12, 48 h and then decreased by 96 h post-ischemia. Immunocytochemistry revealed at 24 h post-ischemia that p65 and p50 immunoreactivity was present in neuronal nuclei of hippocampal CA1 neurons as well as all other hippocampal regions and several other forebrain regions which were not vulnerable to transient forebrain ischemia. At 72 h post-ischemia, nuclear NF-kappa B immunoreactivity had disappeared in all brain areas except in hippocampal CA1 neurons which were degenerating. No evidence for DNA fragmentation as revealed by TUNEL staining could be observed at 24 h. However, at 72 h, hippocampal CA1 neurons were heavily labeled. The results of this study demonstrate that global forebrain ischemia causes a transient activation of NF-kappa B in many forebrain regions. NF-kappa B remains persistently activated in the vulnerable hippocampal CA1 sector. Because of the persistent activation of NF-kappa B in these neurons, the possibility exists that NF-kappa B has a role in programmed cell death in hippocampal CA1 neurons. PMID- 9332716 TI - Angiotensin II type-2 (AT2) receptor-mediated inhibition of NMDA receptor signalling in neuronal cells. AB - The N-methyl-D-aspartate (NMDA) receptor has been reported to be important in synaptic plasticity, neuronal development, normal brain function and neurologic disease. We have recently shown that PC12W cells, a subclone of rat pheochromocytoma PC12 cell line, release nitric oxide (NO), as measured by in vitro spin-trapping combined with electron paramagnetic resonance (EPR) spectroscopy, when challenged with NMDA [Norby, S.W., Weyhenmeyer, J.A. and Clarkson, R.B., Stimulation and inhibition of NO production in macrophages and neuronal cells as observed by spin trapping, Free Rad. Biol. Med., 22 (1997) 1 9]. In the present study, we provide immunochemical evidence for the expression of both the NMDAR1 and NMDAR2A/B receptor subunits in PC12W cells, that express only the angiotensin type-2 (AT2) receptor subtype, and in NG108-15 (NG108) cells, a murine neuroblastoma x glioma hybrid that expresses both the angiotensin type-1 (AT1) and AT2 receptor subtypes. We also show that treatment of PC12W cells with angiotensin (Ang II) decreases NMDA-induced NO release by 28.0 +/- 4.2%, and that this response can be attenuated by pre-treating the cells with the isoform-specific AT2 antagonist, PD 123319. Interestingly, there was no effect on cGMP accumulation in PC12W cells treated with NMDA. Similar experiments were carried out using NG108 cells since the binding properties and functional characteristics of their NMDA receptors have been previously described [Ohkuma, S., Katsura, M., Chen, D., Chen, S. and Kuriyama, K., Presence of N-methyl-D aspartate (NMDA) receptors in neuroblastoma x glioma hybrid NG 108-15 cells analysis using 45Ca2+ influx and [3H]MK-801 binding as functional measures, Mol. Brain Res. 22 (1994) 166-172]. Our results show that NG108 cells significantly increase cGMP levels when challenged with NMDA (21.2 +/- 5.0% over control levels), and that this response can be attenuated by the addition of angiotensin (57.1 +/- 6.2% of stimulated levels). The effect of angiotensin on NMDA-mediated changes in cGMP levels was blocked by the AT2 antagonist, PD 123319, but was not significantly changed by the addition of the AT1 antagonist, losartan. Further, Ang II action on NMDA signalling in NG108 cells was completely inhibited by the addition of both the AT1 and AT2 antagonists. Taken together, these results suggest that AngII inhibits NMDA-mediated NO and cGMP production through a mechanism involving the AT2 receptor subtype. PMID- 9332717 TI - Expression and characterization of human beta-secretase candidates metalloendopeptidase MP78 and cathepsin D in beta APP-overexpressing cells. AB - Human beta-secretase candidates, MP78 (h-MP78, EC 3.4.24.15) and cathepsin D (Cat D, EC 3.4.23.5), were evaluated for their ability to enhance amyloid-beta-protein (A beta) secretion when overexpressed in beta APP-containing cells. HEK-293 cells stably co-expressing h-MP78 or Cat D and h-beta APP695 were metabolically labeled with [35S]methionine and A beta secretion was quantified in the conditioned media by immunoprecipitation and ELISA without showing any significant increase in A beta production. Because Cat D is known to have a higher affinity for APP substrate containing the Swedish familial Alzheimer's disease double mutation (SFAD, K595N and M596L substitutions in beta APP695) than for the wild type substrate [Dreyer et al., Eur. J. Biochem., 224 (1994) 265-271], the effect of Cat D overexpression was tested in a HEK293/beta APPSFAD stable cell line. ELISA analysis of the conditioned media from these cells did also not reveal any increase in A beta generation. In addition, recombinant h-MP78 purified from E. coli cleaved an APP-derived substrate spanning the beta-secretase site (ISEVKMD1AEFRHDS) at multiple sites, but the beta-site cleavage was only a minor one; cleavage occurred predominantly at K-M and E-F bonds. Human liver Cat D also cleaved the same substrate at multiple sites, yet the major cleavage at pH 4.0 occurred at the amyloidogenic D1 site. These findings indicate that h-MP78 does not have the cleavage specificity required for a beta-secretase protease and although Cat D fulfilled the amyloidogenic cleavage specificity, the results of the co-expression experiments make both enzymes less likely candidates as relevant beta-secretases. PMID- 9332718 TI - Characterization of rat C5a anaphylatoxin receptor (C5aR): cloning of rat C5aR cDNA and study of C5aR expression by rat astrocytes. AB - Complement system activation within the central nervous system (CNS) is involved in demyelinating and neurodegenerative disorders, but the role of complement in the pathogenic process or in the repair remains unclear. Besides the direct lytic effects of complement on target cells (oligodendrocytes or neurons), complement can exert other functions through interaction of complement fragments with specific receptors. The C5a anaphylatoxin, an inflammatory peptide which is formed during complement activation, might play a role in the CNS pathogenesis, and activation and recruitment of glial cells by binding to its receptor (C5aR) on CNS cells. Using degenerate primers corresponding to homologous regions between human and mouse C5aR cDNAs, we have cloned a rat C5aR cDNA probe from rat monocytes RNAs after RT-PCR experiment. The rat C5aR probe isolated by this procedure allowed us to clone the rat C5aR cDNA-coding sequence using a library screening cloning strategy. This probe was also used to study the expression of the C5aR mRNA in the rat CNS. Northern blotting and RT-PCR experiments demonstrated the constitutive expression of C5aR mRNA in brain, spleen, kidney and lung. This transcript was also observed in primary culture of rat astrocytes. Microfluorimetry experiments demonstrated that C5aR expressed by astrocytes in culture is functional since the addition of C5a induced a dose-dependent increase of intracellular calcium concentration. The expression of the C5aR by astrocytes suggests new roles for the C5a anaphylatoxin in reactive astrogliosis to CNS injuries. PMID- 9332719 TI - Induction of c-fos and junB mRNA following in vivo brain irradiation. AB - Although radiotherapy is a front line treatment for brain tumors, little is known about the in vivo molecular responses of brain to irradiation. In this study, expression of c-fos, c-jun and junB immediate-early genes were followed in mouse brain after irradiation. C-fos and junB, but not c-jun, mRNA was induced within 15 min in unanesthetized irradiated mice. Induction was transient and lasted < 4 h. The response was dose-dependent with increases in c-fos and junB mRNA levels after dose of > or = 2 and 7 Gy, respectively. Anesthesia of mice with pentobarbitol delayed the increases in mRNA expression and the response was attenuated. Pre-treatment of mice with dexamethasone, in a schedule which suppressed acute-phase gene expression after brain irradiation, did not significantly change c-fos and junB induction. Our results show that c-fos and junB responses occur in the brain in response to irradiation and that they can be modified by pentobarbital treatment but suggest that there is no direct correlation between the level of mRNA expression and later expression of cytokines or other acute-phase response genes. PMID- 9332720 TI - Sequence and expression patterns of two forms of the middle molecular weight neurofilament protein (NF-M) of Xenopus laevis. AB - The middle molecular weight neurofilament protein (NF-M) is relevant to our understanding of vertebrate neurofilaments in growing axons, both because it exists in all vertebrates and because it undergoes characteristic changes in its phosphorylation state during axonal development. Indeed, all vertebrate neurofilament proteins are believed to have originated by gene duplication from an ancestral, NF-M-like protein. The role of NF-M in axonal development has been studied extensively in the frog, Xenopus laevis, through the use of monoclonal antibodies. To acquire a better understanding of the relationship of X. laevis NF M to that of other vertebrates and to obtain additional reagents to study and perturb neurofilaments in developing axons, we isolated cDNA clones from the nervous system. These clones encoded two forms of NF-M, which exhibited 93% amino acid identity overall and 94%, 96% and 90% identity over their head, rod, and tail domains, respectively. Synonymous nucleotide substitution rates between the two forms tied their origin to an ancestral duplication of the Xenopus genome, which occurred approximately 30 million years ago. Non-synonymous substitution rates indicated that the tail domain is evolving more rapidly than the rod domain. Both forms shared structural features in common with other vertebrate NF Ms but had only a single example of the KSP phosphorylation motif that is repeated multiple times in the NF-Ms of bird, goldfish and mammal. In post metamorphic frogs, the NF-M(1) transcript was consistently expressed at higher levels than that of NF-M(2), although their anatomical patterns of expression were qualitatively similar. During development, however, only NF-M(2) was detectable in retinal ganglion cells through stage 42. We speculate that the differences observed between these two forms may represent early stages of protein diversification akin to what occurred after the gene duplications that gave rise to other vertebrate neurofilament proteins. PMID- 9332721 TI - Time course of striatal, pallidal and thalamic alpha 1, alpha 2 and beta 2/3 GABAA receptor subunit changes induced by unilateral 6-OHDA lesion of the nigrostriatal pathway. AB - Immunocytochemical techniques were used to investigate the distribution and abundance of GABAA receptor subunits (alpha 1, alpha 2 and beta 2/3) in the brains of unilaterally 6-OHDA-lesioned rats. Three and 7 days after lesion, the alpha 2-subunit was significantly more abundant in the lesion-ipsilateral striatum than in the lesion-contralateral striatum; by 4 weeks after lesion, however, no significant between-side differences were observed. Three and 7 days after lesion, the alpha 1-subunit was significantly less abundant in the lesion ipsilateral globus pallidus than in the lesion-contralateral side; again, this difference disappeared within 4 weeks of lesion. Similarly, alpha 1 was initially less abundant in several relay thalamic nuclei on the lesioned side while alpha 2 was initially more abundant in intralaminar thalamic nuclei on the lesioned side. There were no significant between-side changes for the beta 2/3-subunits. Comparison of non-lesioned and 6-OHDA-lesioned rats revealed significant differences in brain areas which also showed differences on comparison of the lesioned and non-lesioned sides of 6-OHDA-lesioned rats. These results suggest that there is an early adaptation to the lesion, achieved through changes in GABAA receptor abundance. That some of these changes are no longer apparent after 4 weeks is due not only to partial reversion of the changes in the lesioned side but also to compensatory changes in the non-lesioned side. PMID- 9332722 TI - Glucocorticoids elevate GTP cyclohydrolase I mRNA levels in vivo and in PC12 cells. AB - GTP cyclohydrolase I (GTPCH) is the rate-limiting enzyme in the formation of tetrahydrobiopterin, the cofactor for catecholamine, indolamine and nitric oxide biosynthesis. The effect of glucocorticoids on GTPCH gene expression was examined by direct infusion of cortisol to rats and by incubation of PC12 cells with glucocorticoids. Northern blot analysis revealed that infusion of cortisol for 1 or 7 days elevated levels of the 3.6 kb GTPCH mRNA species in rat adrenal medulla, while the 1.2 kb mRNA species were only increased by 1 day cortisol. Cortisol administration to hypophysectomized animals elicited a 4-5-fold elevation in both forms of GTPCH mRNA. These results indicate that glucocorticoids may be directly involved in the regulation of adrenomedullary GTPCH mRNA levels by physiological stress. Incubation of PC12 cells with plasma from immobilized, but not control, animals increased the level of the 3.6 kb mRNA. Treatment of PC12 cells with dexamethasone for 12-48 h elicited a 4-6-fold elevation in both GTPCH mRNAs. Using the nuclear run-on assay, increased transcription of the GTPCH gene was observed in the rat adrenal medulla with immobilization stress, or in PC12 cells treated with dexamethasone. This is the first report that glucocorticoids can alter GTPCH expression. PMID- 9332723 TI - Expression of group one metabotropic glutamate receptor subunit mRNAs in neurochemically identified neurons in the rat neostriatum, neocortex, and hippocampus. AB - Metabotropic glutamate receptors (mGluRs) can be divided into three groups based on sequence homology and pharmacology. We studied expression of group I mGluRs (mGluR1 and mGluR5) in identified neurons of the rat neostriatum, neocortex, and hippocampus using in situ hybridization. Tissue sections were hybridized with radiolabeled RNA probes for mGluR1 or mGluR5 and digoxygenin labeled RNA probes detecting somatostatin (SOM), preproenkephalin (ENK), preprotachykinin (SP), glutamic acid decarboxylase 67 (GAD67), parvalbumin (PARV), or choline acetyltransferase (ChAT) mRNA. In the striatum, mGluR1 hybridization signal was observed in all six neuronal populations. The strongest signal was found in SP positive neurons, with a lower signal in ENK-positive neurons. All striatal interneurons were labeled less intensely than ENK- and SP-positive projection neurons. For striatal mGluR5 mRNA, both SP- and ENK-positive projection neurons were intensely labeled, but only GAD67-positive interneurons exhibited a significant signal. In the neocortex and hippocampus, mGluR1 and mGluR5 hybridization signals were studied in SOM-, GAD67-, and PARV-positive neurons. Hybridization signal for mGluR1 mRNA was intense in SOM-positive neurons of the cortex, CA1, CA3, and dentate gyrus, and weaker in GAD67-positive neurons of CA3 and dentate gyrus. MGluR5 signals were intensely labeled in SOM-, GAD67- and PARV positive neuronal populations of the cortex and hippocampus. SOM-positive neurons were more intensely labeled in the hippocampus than cortex. PMID- 9332724 TI - Functional expression of olfactory-adrenergic receptor chimeras and intracellular retention of heterologously expressed olfactory receptors. AB - Replacing the G-protein-coupling domains of the beta 2-adrenergic receptor with homologous domains of putative olfactory receptors produced chimeric receptors which were able to stimulate pigment dispersion in Xenopus melanophores, a G protein-mediated pathway. A multiple replacement chimera containing the second, third and C-terminal cytoplasmic domains of receptor OR5 elevated cyclic adenosine 3':5'-monophosphate (cAMP) and suppressed production of inositol phosphates. Co-expression of G alpha olf did not alter the strength of response of this chimera. A novel rat olfactory receptor cDNA (U131) was isolated and sequenced. Expression of U131 and OR5 constructs containing an N-terminal epitope tag or C-terminal fusion to green fluorescent protein occurred in an intracellular network but not in the plasma membrane of heterologous cells. Similarly treated beta 2-adrenergic receptors were functional and were observed in the plasma membrane and the intracellular network. These results demonstrate that the putative cytoplasmic domains of olfactory receptors are capable of functional interaction with heterologous G-proteins of the G alpha s subtype. Instead, the absence of these receptors from the plasma membrane of heterologous cells appears to explain our inability to determine if odorants can activate the olfactory receptor clones. We hypothesize that the olfactory receptors have requirements for maturation and targeting to the plasma membrane that are different from most other G-protein-coupled receptors. PMID- 9332726 TI - Increased gene expression of neuronal nitric oxide synthase in brain of adult spontaneously hypertensive rats. AB - Neuronal nitric oxide is hypothesized to participate in regulation of autonomic function by decreasing sympathetic output to the periphery. This hypothesis predicts that gene expression of neuronal nitric oxide synthase is increased during states of heightened sympathetic activity. To test the hypothesis, we measured gene expression in the spontaneously hypertensive rat (SHR), a genetic model of hypertension in which sympathetic activity is correlated with increasing pressure. SHRs and two strains of control rats (Wistar-Kyoto [WKY] and Sprague Dawley [SD]) at 4 weeks (pre-hypertensive) and 14 weeks (established hypertension) of age were used to measure gene expression in hypothalamus, dorsal pons, dorsal medulla, rostral ventrolateral medulla, and caudal ventrolateral medulla. Semi-quantitative reverse transcription-polymerase chain reactions and in situ hybridization were used to measure changes in neuronal nitric oxide synthase mRNA. No significant differences were found in any of the areas studied among the three strains of rats in the 4-week rats. At 14 weeks significant increases in gene expression were found in the hypothalamus (73% compared to WKYs, 104% compared to SDs), dorsal medulla (31% and 45%), and caudal ventrolateral medulla (24% and 27%) of SHRs. In situ hybridization revealed that neurons expressing the synthase gene in the hypothalamus were found primarily in the paraventricular (both parvo- and magnocellular divisions) and supraoptic nuclei. These data show that gene expression of neuronal nitric oxide synthase is increased in central autonomic centers in animals with increased sympathetic activity and they support the hypothesis that nitric oxide plays an important role in maintenance of homeostatic balance through modulation of sympathetic activity. PMID- 9332725 TI - Functional organization of the promoter region of the mouse F3 axonal glycoprotein gene. AB - F3 is a developmentally regulated adhesive glycoprotein expressed by subpopulations of central and peripheral neurons which mediates neurite growth and fasciculation via cis- and trans-interactions with cell-surface or matrix components. We previously reported on the characterization of the F3 gene 5' flanking region in which we identified promoter and enhancer elements. Here, we report on the functional organization of the F3 gene regulatory regions. We show that the F3 promoter is built of linearly arranged positive and negative elements scattered through the 5' flanking region of the F3 gene and the 1st exon (exon 0). Neural- and cell type-specific expression of F3 appears to be governed by elements located in the most proximal promoter region which includes a neural specific enhancer. In retardation assays, all these cis-acting elements bind nuclear proteins, three of which interact with the identified enhancer element while a single species interacts with sequences located within exon 0. Some of these proteins are also specifically expressed within the brain, indicating that they could correspond to neural-specific trans-acting factors. Elements located immediately upstream of the cell type-specific enhancer and within exon 0 are responsible for regulation of F3 expression by cAMP and retinoic acid. PMID- 9332727 TI - Detection of opioid receptor mRNA by RT-PCR reveals alternative splicing for the delta- and kappa-opioid receptors. AB - The three mu-, delta- and kappa-opioid receptors have recently been cloned and characterized at the molecular level. Our analysis of opioid receptor transcripts by RT-PCR revealed two PCR products derived from delta and kappa mRNAs with size higher than expected from the known cDNA sequences. DNA sequencing showed additional nucleotides inserted between the known splice sites, indicating the possible existence of alternative splicing pathways for delta and kappa receptors. The novel delta-opioid receptor transcript is expressed in mouse brain and contains a 243 bp insertion. This additional sequence is located at the splice junction between the first and second coding exons and is encoded by a single exon located 9 kb upstream exon 2 in the mDOR gene. The other alternative transcript occurs in human monocytic and T lymphocytic cell lines and encodes a novel form of the kappa-opioid receptor. The PCR product presents a 23 bp deletion at the 3' end of exon 2 followed by a 246 bp insertion found between exons 2 and 3. In the hKOR gene, this insertion is encoded by two DNA segments. One of them is located 0.4 kb downstream exon 2 while the second is flanking exon 3 on the 5' side. Both novel putative delta and kappa exons present in-frame stop codons that would lead to truncated receptor proteins. A possible functional or regulatory role of these shorter proteins in opioid function remains to be determined. PMID- 9332728 TI - In vitro activation of the mouse mid-sized neurofilament gene by an NF-1-like transcription factor. AB - In vitro transcription using nuclear extracts from rat brain and liver were used to assess the tissue-specific and functional elements of the mouse neurofilament mid-sized gene promoter (pNF-M). Deletion from -2.7 to -103 (relative to the start site of transcription) resulted in a small increase (2-fold) in the activity of the NF-M promoter in both extracts. Promoter strength was slightly higher in brain vs. liver extracts. Deletion to -49 resulted in a 10-fold loss of promoter activity in brain extracts and 6-fold drop in liver. Transcription in both extracts was TATA box-dependent. The region between -65 and -40 was shown to contain sequences responsible for high-level NF-M promoter activity in brain and liver extracts. Within this region are Sp1 and NF-1-like binding sites. Mutation of the NF-1-like site (-53/-39) caused a large drop in the activity of the NF-M promoter while mutation of the Sp1 site (-64/-57) possibly slightly diminished promoter activity in brain and liver extracts. Both the Sp1 and NF-1-like sites were shown by gel shift competition and supershift assays to be able to bind their respective factors. We conclude that the basic mouse NF-M promoter is a promiscuous promoter whose activity is modulated by a NF-1-like transcription factor. The lack of tissue specificity in an in vitro system strongly suggests an important role for chromatin structure in the regulation of the mouse NF-M promoter. PMID- 9332729 TI - Transforming growth factor-beta protects human hNT cells from degeneration induced by beta-amyloid peptide: involvement of the TGF-beta type II receptor. AB - Post-mitotic, human neurons (hNT cells) which have a phenotype similar to that of terminally differentiated neurons of the central nervous system were generated by treating the NT2/D1 human teratocarcinoma cell line with retinoic acid. Treatment of both hNT and NT2/D1 cells with 10(-5) M beta-amyloid peptide fragment 25-35 (A beta P) for 24 h resulted in a decrease in cell viability as determined by MTT incorporation and Trypan blue exclusion, and also induced an apoptotic morphology in hNT cells. Pre-treatment of cells for 24 h with 10 ng/ml TGF-beta 1 or 2 before addition of A beta P reduced the apoptotic morphology of hNT cells and increased cell viability in hNT cells, but not in NT2/D1 cells. Results of RT PCR, immunohistochemistry and analysis of receptor cross-linking of [125I]TGF beta 1 to the cell membrane, all showed that the TGF-beta type II receptor is expressed by hNT cells, but not NT2/D1 cells. These results suggest that TGF-beta can protect human, terminally differentiated, TGF-beta type II receptor-positive neurons from A beta P toxicity. We propose that the increased expression of TGF beta in brains of patients with Alzheimer's disease may offer some degree of neuroprotection if neurons also express a functional TGF-beta type II receptor. PMID- 9332730 TI - Comparative developmental profile of the neuropeptide Y Y1 receptor gene and protein in the rat brain. AB - Neuropeptide Y (NPY) is one of the most abundant peptides found in the mammalian central nervous system (CNS) and plays several important roles in regulating brain function. Physiological roles of NPY in the brain are mediated by at least six receptor subtypes (Y1 to Y6). In the present study, a rat Y1 receptor cRNA probe was used for in in situ hybridization experiments in order to determine the developmental profile of this receptor mRNA and to compare it with its translated protein using receptor autoradiography with the radiolabelled ligand [125I][Leu31,Pro34]PYY. The NPY Y1 receptor mRNA is expressed as early as by the 12th day of gestation while specific [125I][Leu31,Pro34]PYY binding is observed by day 14 of gestation. Thereafter, both signals steadily increased, with Y1 receptor mRNA increasing faster than its translated protein during fetal life. The in situ hybridization signals reached a plateau around birth and remained high during the first 2 post-natal weeks to display the adult distribution by the end of the 3rd post-natal week. Similarly, specific [125I][Leu31,Pro34]PYY binding constantly increased during brain maturation and reached a plateau by the end of the 3rd post-natal week. In some brain areas, such as the cerebral cortex, specific binding declined slightly before attaining its adulthood pattern. Throughout ontogenesis, the profile of both the Y1 receptor mRNA and protein was well-matched except in hypothalamic areas where relatively higher mRNA signals were observed. Taken together, these results along with previous reports describing NPY-like immunoreactivity in the early developmental rat brain, suggest that the NPY Y1 receptor may play an important role in early brain development and maturation on the basis of its very early pattern of embryonic expression. PMID- 9332731 TI - Glutamate decarboxylase (GAD65) gene expression is increased by dopamine receptor agonists in a subpopulation of rat striatal neurons. AB - The mRNA levels encoding for the two isoforms of glutamate decarboxylase (GAD65 and GAD67) were measured in the adult rat striatum following systemic administration of dopamine receptor agonists. Double-labeling in situ hybridization histochemistry was used to measure GAD65 or GAD67 mRNA levels in neurons labeled or not with a preproenkephalin (PPE) cRNA probe. Chronic treatment with the D1/D2 dopamine receptor agonist apomorphine or with the D1 dopamine receptor agonist SKF-38393 induced an increase in GAD65 but not GAD67 mRNA levels in different sectors of the striatum. These effects were abolished by pre-administration of the D1 dopamine receptor antagonist SCH-23390. On double labeled sections, GAD65 mRNA labeling was distributed in neurons labeled and unlabeled with the PPE cRNA probe. About half of all neuronal profiles labeled with the GAD65 cRNA probe were also labeled with the PPE cRNA probe. Quantification of labeling at cellular level demonstrated a significant increase of GAD65 mRNA levels in PPE-unlabeled neurons. On the other hand, no significant changes of GAD65 mRNA levels were detected in PPE-labeled neurons. Our results demonstrate a differential effect of dopamine receptor agonists on striatal GAD65 and GAD67 gene expression. In particular, we show that GAD65 mRNA levels are selectively increased in presumed striato-nigral neurons following treatments with dopamine receptor agonists. These data provide evidence that the GAD65 isoform is preferentially involved in the regulation of GABAergic neurotransmission in striato-nigral neurons. PMID- 9332732 TI - Tissue-specific glucocorticoid regulation of tryptophan hydroxylase mRNA levels. AB - A potential long-term target of glucocorticoid modulation of serotonin (5-HT) production is tryptophan hydroxylase (TPH) gene expression. However, studies on TPH gene expression have been hampered by the extremely low levels of TPH mRNA in the brain, and there have been contradictory reports on the effects of glucocorticoids on 5-HT levels. To overcome these obstacles, we have developed a sensitive competitive RT-PCR assay to directly measure TPH mRNA levels from the rat brain. We observed a tissue-specific modulation of TPH mRNA levels in the melatonin producing pineal gland and the serotonin producing raphe nuclei of the brain. Following chronic treatment of adrenalectomized rats with the synthetic glucocorticoid dexamethasone for 1 week, there was a 16-fold increase in TPH mRNA in the pineal gland that was contrasted by a decrease in TPH mRNA to 16% of the control levels in the brain. To address the mechanism of dexamethasone repression of TPH mRNA levels, we then tested a serotonergic neuronal-like cell line derived from rat thyroid C cells. Dexamethasone caused a rapid decrease in TPH mRNA levels to approximately 20% of control values in CA77 C cells. This was measured by both competitive RT-PCR and a standard hybridization assay, which confirmed the validity of the RT-PCR assay. Furthermore, the reduction of TPH mRNA levels was associated with a decrease in 5-HT levels in the CA77 C cells. Hence, glucocorticoids may alter serotonin and melatonin biosynthetic capacity by cell specific modulation of the TPH gene. PMID- 9332733 TI - Effects of neuronal proteoglycans on activity-dependent growth responses of fetal hippocampal neurons. AB - Excitatory amino-acid (EAA) neurotransmitters act as molecular signals influencing the structure of neurons during development. However, the signal transduction and effector mechanisms responsible for these effects have yet to be fully elucidated. We have previously provided evidence that EAA agonists induce the synthesis and release of proteoglycans (PGs) with neurite-promoting activity from fetal hippocampal neurons. In the present studies exposure of fetal hippocampal neurons to glutamate (100 microM) for 5 min resulted in increases in the neuron-specific growth-associated genes T alpha 1 alpha-tubulin (T alpha 1), microtubule-associated protein-2 (MAP-2) and growth-associated protein-43 (GAP 43). mRNA levels peaked at between 8 and 12 h following exposure as determined by competitive reverse transcription polymerase chain reaction (RT-PCR). Increases in neurite growth as measured by axonal length, the total length of dendrites, the number of branches per axon, the total length of branches per axon and the total neurite length were also observed 48 h after glutamate exposure. The increase in T alpha 1, MAP-2 and GAP-43 mRNA levels following glutamate exposure was mediated via both N-methyl-D-aspartate and metabotropic receptor activation. Heparin, which inhibits the neurite growth-promoting effects of PGs in vitro, and heparitinase, which catalyzes the cleavage of heparan sulphate, also inhibited the glutamate-dependent induction of T alpha 1, MAP-2 and GAP-43 mRNA expression and neurite growth when added to culture medium following glutamate exposure. Chondroitin sulphate and chondroitinase AC had no effects on the mRNA levels tested or on neurite growth. Therefore, these studies suggest that neuronal PGs regulated by activation of EAA receptors mediate neuronal growth responses. PMID- 9332734 TI - Topographic comparison of the expression of norepinephrine transporter, tyrosine hydroxylase and neuropeptide Y mRNA in association with dopamine beta-hydroxylase neurons in the rabbit brainstem. AB - In mammalian species, ovulation occurs following a massive release of hypothalamic gonadotropin-releasing hormone (GnRH). Several chemicals, including norepinephrine (NE) and neuropeptide Y (NPY), are responsible for the initiation and/or magnitude and duration of this pre-ovulatory GnRH surge. In the central nervous system, NE neural cell bodies are located in the brainstem; some are co localized with NPY neurons and/or co-express the NE transporter (NET) gene which dictates NET protein production. The activity of NET at NE terminals is critical for synaptic NE function. In the rabbit, coitus induces a hypothalamic NE release which precedes the GnRH surge. We hypothesize that the coital stimulus is transmitted to the brainstem and transformed and integrated into GnRH-stimulating signals via NE, NET and/or NPY. However, very little is known about the distribution of cells expressing NET, NPY and tyrosine hydroxylase (TH, the rate limiting enzyme of NE synthesis) in this species. Therefore, we utilized the sensitive in situ hybridization technique to identify the presence of these messages in conjunction with the location of NE cells, the latter being marked by dopamine beta-hydroxylase (DBH), the specific enzyme for NE synthesis. Three non mated New Zealand White does were perfused with 4% paraformaldehyde and their brainstems were sectioned at 20-micron thick between 2 mm caudal to the obex and the rostral pons. Serial sections were immunohistochemically stained for DBH and hybridized with rabbit-specific TH and NET cRNAs and a human NPY probe. The data suggest that several DBH-positive areas in the medulla expressed one or more messages, i.e. the lateral tegmentum (A1) and the nucleus of the solitary tract (A2) expressed all three mRNAs, the area postrema (AP) contained NET and TH mRNAs but not NPY cells. In the pons, the locus coeruleus (LC), subnucleus of coeruleus (LCs) and lateral tegmental nuclei (A5) expressed NET and TH mRNAs but contained little or no NPY message. The distribution patterns of TH and NET appeared to be similar in the LC, LCs, A2 and AP. PMID- 9332735 TI - Etoposide-induced PC12 cell death: apoptotic morphology without oligonucleosomal DNA fragmentation or dependency upon de novo protein synthesis. AB - Etoposide, a topoisomerase II inhibitor used in cancer therapy, has been shown to induce apoptosis in vitro in a variety of cell types. In the present study, we have characterized the effects of etoposide on undifferentiated rat pheochromocytoma PC12 cells. Etoposide killed PC12 cells in a time- and concentration-dependent manner. 20-24 h incubation with 10 micrograms/ml etoposide induced 25-50% cell death. Hoechst 33258 staining revealed apoptotic morphology in dying cells. No evidence was found of either oligonucleosomal DNA fragmentation, as shown by agarose gel electrophoresis, or endonuclease involvement, as shown by the inability of aurintricarboxylic acid to prevent cell death. Cycloheximide and actinomycin-D were unable to prevent etoposide cytotoxicity indicating that the process is not dependent upon de novo protein or mRNA synthesis. NGF (5 ng/ml) prevented etoposide-induced PC12 cell death. These results offer an example of how the morphological features of apoptosis are not necessarily associated with oligonucleosomal DNA fragmentation or with de novo macromolecule synthesis. PMID- 9332736 TI - Ontogeny of the striatal neurons expressing the D2 dopamine receptor in humans: an in situ hybridization and receptor-binding study. AB - D2 dopamine receptor (D2R) gene expression was analyzed by in situ hybridization and D2R ligand autoradiography in the human striatum during ontogeny. D2R mRNA and ([3H]YM-09151-2)-binding sites were detected in the striatum from week 12 of fetal life. At this time, D2R mRNA and binding sites were predominant in the putamen and occurred in a pattern of clusters. D2R-binding sites displayed a similar pattern. The signal in the caudate nucleus was weak from weeks 12 to 16. From week 20 of fetal life, D2R mRNA and D2R-binding sites signals became intense in the ventral striatum. At birth, D2R mRNA became homogeneously distributed while D2R-binding sites kept an heterogeneously distribution. Comparative topological and temporal analysis of the D2R, enkephalin and D1 dopamine receptor (D1R) mRNAs showed a distinct developmental pattern for each mRNA. Before birth, the neurons expressing enkephalin and D1R mRNAs were preferentially distributed in the matrix and in the striosomes, respectively, while the neurons expressing D2R mRNA did not display a preferential localization. At birth, high levels of enkephalin mRNA were restricted to the matrix; D1R mRNA level was homogeneous throughout the striatum. D2R mRNA was heterogeneously distributed in the whole striatum with high signals located both in the striosomes and the matrix. These results demonstrate that functional D2R are expressed as early as week 12 in the striatum with a heterogeneous distribution. Our findings also demonstrate that, in contrast to what was expected from similar studies in rodents, D2R mRNA and enkephalin mRNA do not display identical, overlapping expression patterns in striatal neurons during human ontogeny. PMID- 9332737 TI - Alterations in BDNF and NT-3 mRNAs in rat hippocampus after experimental brain trauma. AB - Previous studies have suggested that the neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) are neuroprotective or neurotrophic for certain subpopulations of hippocampal neurons following various brain insults. In the present study, the expression of BDNF and NT-3 mRNAs in rat hippocampus was examined after traumatic brain injury. Following lateral fluid percussion (FP) brain injury of moderate severity (2.0-2.1 atm) or sham injury, the hippocampi from adult rats were processed for the in situ hybridization localization of BDNF and NT-3 mRNAs using 35S-labeled cRNA probes at post-injury survival times of 1, 3, 6, 24 and 72 h. Unilateral FP injury markedly increased hybridization for BDNF mRNA in the dentate gyrus bilaterally which peaked at 3 h and remained above control levels for up to 72 h post-injury. A moderate increase in BDNF mRNA expression was also observed bilaterally in the CA3 region of the hippocampus at 1, 3, and 6 h after FP injury, but expression declined to control levels by 24 h. Conversely, NT-3 mRNA was significantly decreased in the dentate gyrus following FP injury at the 6 and 24 h survival times. These results demonstrate that FP brain injury differentially modulates expression of BDNF and NT-3 mRNAs in the hippocampus, and suggest that neurotrophin plasticity is a functional response of hippocampal neurons to brain trauma. PMID- 9332738 TI - The embryonic and post-natal expression of the nicotinic receptor alpha 3-subunit in rat lower brainstem. AB - The mRNA expression of the alpha 3 nicotinic acetylcholine receptor subunit in the rat lower brainstem is more widespread in the embryo and at birth than in the mature brain, but the major cell groups expressing alpha 3 are generally the same, with two exceptions. The transcript for the alpha 3-subunit is transiently expressed in the ventral cochlear nucleus (VCN). Expression was very high in the embryonic and early post-natal VCN, but was significantly decreased in the VCN late in the post-natal period (about post-natal day 15). Conversely, alpha 3 mRNA was not expressed in the motor trigeminal nucleus until about PN3. PMID- 9332739 TI - GABA release and GAD67 mRNA expression in rat hippocampus following entorhinal cortex activation. AB - This study investigate the effect of stimulation of glutamatergic afferents originating in the entorhinal cortex on possible changes of GABAergic transmission in the CA1 subregion of the hippocampus. Microdialysis was used to monitor extracellular GABA and in situ hybridization to measure levels of glutamic acid decarboxylase67 (GAD67) mRNA. A dose-dependent increase in extracellular levels of GABA in the dorsal CA1 subregion was detected following injection of 2.4 and 9.6 micrograms quisqualate into the lateral entorhinal cortex whereas 0.24 microgram had no effect. The GABA increase was attenuated by local administration of tetrodotoxin (TTX), indicating neuronal origin. A 60% decrease and a 160% increase were seen in levels of GAD67 mRNA in the CA1 following injection of 0.24 and 9.6 micrograms quisqualate, respectively. This study provides evidence of an entorhinal cortex influenced stimulatory effect on GABAergic activity in the CA1. However, no direct relationship was found between stimulated GABA release and subsequently measured GAD67 mRNA levels. The increased GABA release and the apparent adaptive increase in GAD67 mRNA levels by the strongest stimulation may be due to an endogenous inhibitory neuroprotective response to an excitotoxic influence. PMID- 9332740 TI - Disease surveillance. PMID- 9332742 TI - Licensure. PMID- 9332741 TI - Animal blood bank. PMID- 9332743 TI - Streaming. PMID- 9332744 TI - An ethicist's commentary on the case of the financially stressed client. PMID- 9332745 TI - The Canadian Animal Health Network. PMID- 9332746 TI - The effect of carotid artery occlusion on lingual arterial blood pressure in dogs. AB - Although temporary occlusion of the carotid arteries is commonly done to reduce blood loss during nasal surgery in the dog, data supporting its use are mostly anecdotal and subjective. Twelve dogs were placed under general inhalation anesthesia and mechanically ventilated to maintain normocapnea and an end-tidal halothane concentration equivalent to 1.3 times the minimum alveolar concentration. Tourniquets were placed around both carotid arteries of each dog. Both lingual arteries were cannulated in each dog and their heart rate and blood pressure were measured bilaterally. During unilateral carotid artery occlusion, the blood pressures in the ipsilateral lingual artery were significantly (P < 0.05) lower than the preocclusion control pressures and pressures recorded in the contralateral vessel. Bilateral carotid artery occlusion resulted in a further significant (P < 0.05) fall in all lingual arterial pressures. The recorded heart rates only varied significantly from preocclusion control values when they increased during bilateral carotid occlusion (P < 0.05). The results of this study confirm that carotid artery occlusion has the potential to reduce intraoperative blood loss during oronasal surgery in the dog. PMID- 9332748 TI - Desmotomy for treatment of chronic desmitis of the accessory ligament of the deep digital flexor tendon in a horse. AB - Chronic lameness was determined to be caused by desmitis of the accessory ligament of the deep digital flexor tendon and adhesions associated with these 2 structures. Desmotomy of the accessory ligament, resection of adhesions, and controlled exercise during convalescence resulted in return to normal use without apparent lameness. PMID- 9332747 TI - A benefit to cost analysis of the effect of premilking teat hygiene on somatic cell count and intramammary infections in a commercial dairy herd. AB - A field trial was conducted to determine the effect of premilking teat disinfection (predipping) on several measures of mastitis in a commercial dairy farm where the predominant organisms isolated from intramammary infections were coagulase negative Staphylococcus spp. Cows were randomly assigned to a treated (predipped with 0.5% iodine germicide plus "good udder preparation") or a control group ("good udder preparation" alone). Sterile composite milk samples were collected at the initiation of the trial and on an approximately bimonthly basis throughout the duration of the trial. There was no difference in the prevalence of isolation of coagulase-negative Staphylococcus spp. from composite milk samples obtained during the 6 herd cultures. The incidence rate for clinical mastitis in the control group was 1.38 cases per 1000 cow days. The incidence rate for clinical mastitis in the treatment group was 1.06 cases per 1000 cow days. The ratio of these 2 was 1.3, suggesting a higher rate in the control group, but the ratio was not statistically significant (P = 0.34). Logistic regression analysis indicated that the effect of treatment group was not significant, although the coefficient suggested that predipping reduced the risk of clinical mastitis. The benefit to cost ratio of 0.37 indicated that the benefit of reduced incidence of clinical cases of mastitis would not have justified the added expense required to predip the herd. PMID- 9332749 TI - Combined hiatal and pleuroperitoneal hernia in a shar-pei. AB - This article presents an unusual combination of a type IV hiatal hernia and a pleuroperitoneal hernia in a young shar-pei. Pathogenesis, diagnosis, and treatment of both conditions are discussed. At surgery, close examination and palpation of the whole diaphragm are recommended to allow perioperative diagnosis of unexpected defects. PMID- 9332750 TI - Squamous cell carcinoma originating from a cutaneous scar in a llama. AB - A nonhealing wound associated with a laceration in a 12-year-old llama was evaluated. Initial attempts at closure were unsuccessful and biopsy revealed scar tissue. Subsequent biopsies, 18 mo later, revealed squamous cell carcinoma with regional metastasis. This report describes squamous cell carcinoma, secondary to a traumatic wound in a llama. PMID- 9332751 TI - An oxytetracycline residue depletion study to assess the physiologically based pharmokinetic (PBPK) model in farmed Atlantic salmon. PMID- 9332752 TI - Presumptive ethylene glycol poisoning in chickens. PMID- 9332753 TI - Epidemiology of dog and cat euthanasia across Canadian prairie provinces. PMID- 9332755 TI - Understanding the epidemiology of genital infection with oncogenic and nononcogenic human papillomaviruses: a promising lead for primary prevention of cervical cancer. PMID- 9332754 TI - The legal implications of the veterinarian's role as a private practitioner and health professional, with particular reference to the human-animal bond: Part 2, The veterinarian's role in society. PMID- 9332756 TI - The lifetime risk of developing prostate cancer in white and black men. AB - Two factors help explain increases in the lifetime risk of developing cancer: (a) decreasing overall mortality rates such that people are now living to older ages when cancer rates rise rapidly; and (b) increasing numbers of cancer cases discovered by new medical procedures, screening tests, and changes in the population risk factors. Prostate cancer lifetime risk estimates are particularly influenced by improved mortality rates and increased detection of asymptomatic disease. In this study, we report trends in lifetime risk estimates of developing prostate cancer in white and black men in the United States, from 1975 to 1993, and focus on the effects of changing mortality and screening. For the study period 1975-1977 to 1991-1993, the lifetime risk of developing invasive prostate cancer increased from 7.3 to 19.6% for whites and from 8.5 to 18.6% for blacks. When we recalculated these estimates using age-specific incidence trends from 1975 through 1989 (thereby controlling for the effect of prostate-specific antigen serum testing on prostate cancer incidence rates), the lifetime risk estimates in 1991-1993 fell to 13.8% for whites and 12.5% for blacks. When we made an additional assumption, basing lifetime risk estimates on higher 1975-1977 mortality rates, the lifetime risk estimates in 1991-1993 became 11.3% for whites and 11.8% for blacks. It is also shown that although mortality rates have improved for white and black men over the study period, they are much larger for blacks than whites in younger age groups, when the prevalence of prostate cancer is relatively low. As a result, fewer blacks survive to older ages when age specific prostate cancer rates are large. It is of note that blacks have higher incidence rates for prostate cancer than do whites at every age-specific interval. Hence, increasing trends in lifetime risk of prostate cancer suggest, in large part, longer life expectancy and better detection methods. PMID- 9332757 TI - Relationship between vitamin and calcium supplement use and colon cancer. AB - The relationship between vitamin supplement use and colon cancer was assessed in a population-based case-control study among men and women aged 30-62 years. Cases were 251 men and 193 women diagnosed with colon cancer in 1985-1989 in three counties in the Seattle metropolitan area who were identified from the Surveillance, Epidemiology, and End Results cancer registry. Controls were 233 men and 194 women identified by random digit dialing. Supplement use was assessed by questions on frequency, duration, and dose per day (for individual supplements) or type (for multivitamins) during the 10-year period ending 2 years before diagnosis. All results were adjusted for age and sex and were not confounded by other measured behaviors. The average daily intake of supplemental vitamins A, C, E, folic acid, calcium, and multivitamins during the reference period were each associated with reduced risk of colon cancer (all P for trend < 0.03). The strongest associations were for use of vitamin E (odds ratio, 0.43; 95% confidence interval, 0.26-0.71 for > or = 200 IU/day versus none) and multivitamins (odds ratio, 0.49; 95% confidence interval, 0.35-0.69 for daily use versus no use; both P for trend < 0.001). These two associations were also significant using a stricter test of trend limited to supplement users, which reduces the effect of colinearity among these exposures. Because almost all vitamin D supplementation comes from multivitamin pills, the association of vitamin D use with colon cancer could not be distinguished from that of multivitamin use. Clinical trials or cohort studies with long-term assessment would be needed before public health recommendations could be made about supplement use. PMID- 9332758 TI - Alcohol consumption in relation to endometrial cancer risk. AB - We analyzed data from a population-based case-control study of Wisconsin women to evaluate the relationship of alcohol consumption to endometrial cancer risk. Cases (n = 739) were identified from a statewide tumor registry; controls (n = 2313) were selected randomly from driver's license lists and Medicare beneficiary files. Alcohol consumption and other factors were ascertained by telephone interview. Compared with abstainers, the multivariable relative risk for recent consumption of two or more drinks per day was 1.27 [95% confidence interval (CI) 0.78-2.07]; increasing consumption was not associated with risk of disease (P for trend, 0.82). The relative risk for early adulthood consumption of two or more drinks per day was 1.00 (95% CI, 0.58-1.73), with no suggestion of a trend (P = 0.26). Although the sample size was limited, a significant inverse association was suggested in premenopausal women consuming one drink per day or more (0.20, 95% CI 0.06-0.71). Beverage-specific consumption was not associated with risk. This study suggests that, unlike breast cancer, endometrial cancer is not positively associated with alcohol intake. PMID- 9332759 TI - Association of adenocarcinoma and squamous cell carcinoma of the esophagus with tobacco-related and other malignancies. AB - Little is known about the etiology of esophageal and gastric cardia adenocarcinoma (EGA), a cancer with one of the fastest-rising incidences in the developed world. To explore the etiology of this cancer, we conducted a retrospective cohort analysis using data from the Surveillance, Epidemiology and End Results Program of the United States National Cancer Institute to study EGA and esophageal squamous cell carcinoma (ESC), in association with cancers of other sites. Standardized incidence ratios, adjusted for age, sex, and time period, were calculated as a measure of the relative risk (RR) of developing a second primary cancer (EGA or ESC) following a given first primary site. We found a moderately elevated risk of EGA following cancers of the lung (RR = 1.9 in men and RR = 2.0 in women) and of the head and neck (RR = 2.1 in men and RR = 6.3 in women) and a strongly elevated risk of ESC following cancers of the lung (RR = 2.8 in men and RR = 5.1 in women) and of the head and neck (RR = 9.6 in men and RR = 38.8 in women). A significantly elevated risk following breast cancer in women was observed for both EGA (RR = 2.6; 95% confidence interval, 1.8-3.7) and ESC (RR = 1.4; 95% confidence interval, 1.1-1.9). We also found a significantly elevated risk of EGA following bladder (RR = 2.0), colorectal (RR = 1.7), and prostate (RR = 1.4) cancer in men and of ESC following colorectal cancer (RR = 1.7) in women in this study. The strong association with tobacco-related malignancies in this study reinforces the role of tobacco in the etiology of esophageal cancers, which appears stronger for squamous cell carcinoma than for adenocarcinoma and stronger in women than in men. The study also suggests a possible shared etiology between esophageal adenocarcinoma and colorectal cancer in men and provides new evidence about the association of both adenocarcinoma and squamous cell carcinoma of the esophagus with breast cancer in women. Findings of this study provide clues to the etiology of EGA and ESC. PMID- 9332760 TI - Differences in the urinary metabolites of the tobacco-specific lung carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone in black and white smokers. AB - Incidence and mortality rates for lung cancer in the United States are significantly greater in blacks than in whites. This disparity cannot be explained by differences in smoking behavior. We hypothesize that the observed racial differences in risk may be due to differences in the metabolic activation or detoxification of the tobacco-specific lung carcinogen 4-(methylnitrosamino)-1 (3-pyridyl)-1-butanone (NNK). To test this, different biomarkers of NNK exposure and metabolism, including the urinary metabolite 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanol (NNAL) and the presumed detoxification product [4 (methylnitrosamino)-1-(3-pyridyl)but-1-yl]-beta-O-D-glucosiduronic acid (NNAL Gluc), were examined along with questionnaire data on lifestyle habits and diet in a metabolic epidemiological study of 34 black and 27 white healthy smokers. Results demonstrated that urinary NNAL-Gluc:NNAL ratios, a likely indicator of NNAL glucuronidation and detoxification, were significantly greater in whites than in blacks (P < 0.02). In addition, two phenotypes were apparent by probit analysis representing poor (ratio < 6) and extensive (ratio > or = 6) glucuronidation groups. The proportion of blacks falling into the former, potentially high-risk group was significantly greater than that of whites (P < 0.05). The absolute levels of urinary NNAL, NNAL-Gluc, and cotinine were also greater in blacks than in whites when adjusted for the number of cigarettes smoked. None of the observed racial differences could be explained by dissimilarities in exposure or other sociodemographic or dietary factors. Also, it is unlikely that the dissimilarities are due to racial differences in preference for mentholated cigarettes, because chronic administration of menthol to NNK-treated rats did not result in either increases in urinary total NNAL or decreases in NNAL-Gluc:NNAL ratios. Altogether, these results suggest that racial differences in NNAL glucuronidation, a putative detoxification pathway for NNK, may explain in part the observed differences in cancer risk. PMID- 9332761 TI - CYP1A1 and GSTM1 polymorphisms and oral cancer risk. AB - The importance of both the CYP1A1 exon 7 (ile:val) and GSTM1 (0/0) polymorphisms in oral cancer susceptibility was assessed by examining polymorphic prevalences in 135 patients with oral cancer and 135 noncancer controls frequency-matched by age at diagnosis (+/- 5 years), race, sex, and institute of patient recruitment. The prevalence of the GSTM1 (0/0) genotype was approximately 51% in both cases and controls. The prevalence of the CYP1A1 (ile:val) polymorphism [including both the (ile/val) and (val/val) genotypes] was significantly higher in cases as compared to controls (17.6% versus 7.6%, respectively; crude odds ratio, 2.6; confidence interval, 1.2-5.7). No association was observed between polymorphic prevalence and levels of smoking or alcohol consumption in cases. These results suggest that the GSTM1 null genotype is not associated with oral cancer risk. These results also suggest that individuals with the CYP1A1 exon 7 ile:val polymorphism are at increased risk for oral cancer, and that this risk may not be influenced by differences in exposure to tobacco smoke. PMID- 9332762 TI - Determinants for genital human papillomavirus (HPV) infection in 1000 randomly chosen young Danish women with normal Pap smear: are there different risk profiles for oncogenic and nononcogenic HPV types? AB - Most studies of risk factors for human papillomavirus (HPV) DNA detection have focused on overall HPV positivity and have not examined determinants for high risk and low-risk HPV types separately. We studied risk determinants for genital HPV infection in 1000 randomly chosen women (20-29 years) with normal cervical cytology from Copenhagen, Denmark. All women had a personal interview, a Pap smear, and cervical swabs for HPV DNA detection using a PCR technique. On the basis of their association with cervical cancer, the HPV types were categorized as belonging to a high-risk group ("oncogenic types") or a low-risk group ("nononcogenic types"). The overall HPV detection rate was 15.4%. Of HPV-positive women, 74% had oncogenic HPV types, and 30% had nononcogenic HPV types. Younger age and lifetime measures of sexual activity (notably, number of partners) were the main risk factors for the oncogenic HPV types. Furthermore, a previous Chlamydia infection was associated with the high-risk HPV types. In contrast, the most important determinants for nononcogenic HPV infection were contraceptive variables related to the physical protection of the cervix (condom or diaphragm) and number of partners in the last 4 or 12 months. Our study confirms the venereal nature of HPV infection. We hypothesize that the low-risk HPV infection, which correlates with recent sexual behavior, may be more transient than infection with the oncogenic HPV types, which correlates with lifetime exposure measurements of sexual habits. PMID- 9332763 TI - Immune activation in cervical neoplasia: cross-sectional association between plasma soluble interleukin 2 receptor levels and disease. AB - In a previous study (Tsukui et al., Cancer Res., 56: 3967-3974, 1996), we observed an inverse association between degree of cervical neoplasia and interleukin (IL) 2 production by peripheral blood mononuclear cells in response to human papillomavirus (HPV) 16 E6 and E7 peptides in vitro. This suggested that a Th1-mediated cellular immune response might be important in host immunological control of HPV infection and that a lack of such a response might predispose to progression of cervical disease. To follow up on these findings, we have conducted a cross-sectional study of women with various degrees of cervical neoplasia to investigate the association between overall immune activation and cervical disease. A total of 235 women were recruited into our study; 120 of these women were participants in our previous study in which IL-2 production in response to HPV-16-specific peptides was measured. The study population included 34 women with invasive cancer, 62 women with high-grade squamous intraepithelial lesions (HSILs), and 105 women with low-grade squamous intraepithelial lesions (LSILs). In addition, 34 cytologically normal women with no past history of squamous intraepithelial lesions despite confirmed HPV-16 infection in the 5 years preceding the study were selected as controls. As our measure of overall immune activation, serum samples obtained from study participants were tested for soluble IL-2 receptor (sIL-2R) level using an ELISA method. The mean sIL-2R levels were found to increase with increasing disease severity (Ptrend = 0.0002). Among cytologically normal, HPV-exposed women, the mean receptor level in serum was 465.8 units/ml compared to 467.6 units/ml among LSIL subjects, 514.9 units/ml among HSIL subjects, and 695.5 units/ml among women with invasive cervical cancer. Similarly, the proportion of women with elevated sIL-2R levels (defined as > or = 450 units/ml) increased with increasing disease severity from 35.2% among normal study subjects to 70.6% among cancer patients (Ptrend = 0.003). Among the subgroup of subjects for whom in vitro IL-2 production in response to HPV-16-specific peptides was measured, we examined the association between in vitro IL-2 production and serum levels of sIL-2R. sIL-2R levels were higher, on average, among those women who were positive in our IL-2 production assay compared to those who were negative, but the differences did not reach statistical significance (P > 0.05). We also observed a trend of increasing sIL 2R level with increasing disease severity both in women who were positive and in women who were negative for our IL-2 production assay, but the trend was only significant among those who were negative for IL-2 production (Ptrend = 0.01). Results from our studies suggest that although the immune system of women with cervical neoplasia is nonspecifically activated as disease severity increases, the ability of those women with HSILs or cancer to mount a Th1-mediated immune response to HPV peptides appears to decrease compared to women with LSILs or normal women infected with HPV. Increased overall activation along with decreased Th1 immune response among women with increasing cervical disease severity might be explained by an increased Th2-mediated immune response, a response that we hypothesize is ineffective in controlling the viral infection and its early cytological manifestations. Future studies should directly assess Th2-mediated responses to confirm this hypothesis. Also, future efforts should be aimed at determining whether the associations observed are causally related to disease progression or an effect of the disease. PMID- 9332764 TI - A case-case analysis of factors related to overexpression of p53 in endometrial cancer following breast cancer. AB - We studied 54 patients diagnosed with endometrial cancer between 1981 and 1994 following a diagnosis of breast cancer. We used a case-case analysis, comparing tumors with and without overexpression of the p53 gene product to evaluate the association of putative p53 mutations with tamoxifen use and other risk factors for endometrial cancer. Twenty-four % of the tumors showed strong positive staining for the p53 gene product. Tumors in a more advanced stage (stage 2, 3, or 4, compared to stage 1) were more likely to overexpress p53 [odds ratio (OR) = 4.2; 95% confidence interval (CI), 1.1-16.2], as were tumors with serous or clear cell, compared to endometrioid, histology (OR = 5.8; 95% CI, 1.3-26.5). There was a small association between p53 overexpression and treatment with tamoxifen for breast cancer (OR = 2.6; 95% CI, 0.69-9.8). There was a strong relationship between overexpression of p53 and having a first-degree relative with breast cancer (OR = 12.3; 95% CI, 2.6-57.4) and between overexpression of p53 and having an additional cancer, i.e., at sites other than breast or endometrium (OR = 7.9; 95% CI, 1.6-40.1). In this group of women, genetic predisposition to cancer, as reflected in family history of breast cancer and personal history of an additional primary cancer, was strongly associated with overexpression of p53 in endometrial tumors. The results suggest that use of tamoxifen may be associated with an increase in tumors that overexpress p53, although the results could be due to chance. PMID- 9332765 TI - The effect of incubation of rectal biopsies on measures of proliferation using proliferating cell nuclear antigen in comparison with 5-bromo-2-deoxyuridine. AB - Rectal epithelial cell kinetics are used as intermediate markers for colorectal cancer and relate to risk. In this study, measures of proliferation using direct immunohistochemistry for proliferating cell nuclear antigen (PCNA) were compared to in vitro labeling by bromodeoxyuridine (BrdUrd) and incubated biopsies that were later stained for PCNA (PCNA-I) in human rectal biopsies. The study group consisted of 20 sets of biopsies from 12 subjects participating in an intervention trial. Fresh nonincubated biopsies were fixed in methacarn and stained immunohistochemically for PCNA (clone 19A2). In parallel biopsies, BrdUrd was incorporated into the DNA of S-phase cells during a 2-h incubation at 37 degrees C under hyperbaric conditions and localized by immunohistochemistry. Additionally, biopsies were incubated under hyperbaric conditions for 2 h at 37 degrees C, fixed in methacarn, and stained for PCNA (PCNA-I). There was a highly significant difference in the labeling index between the three methods (P < 0.01), but there was no significant difference between subjects (P = 0.439). The mean labeling index was 2.3 +/- 0.1% for PCNA, 2.9 +/- 0.1% for PCNA-I, and 4.1 +/- 0.1% for BrdUrd. The proportion of labeled cells in the top two-fifths was significantly higher (P = 0.01) for BrdUrd (5.5 +/- 0.8%) and PCNA-I (6.4 +/- 1.1%) compared to PCNA (3.1 +/- 0.6%), and a significant difference was seen between subjects (P = 0.038). PCNA-I and BrdUrd methods had similar crypt heights with 73.5 +/- 1.8 and 71.2 +/- 1.3 cells/crypt column, respectively, but were significantly shorter (P < 0.001) than PCNA with 83.4 +/- 1.5 cells/crypt column, indicating a loss of cells during organ culture. The simplicity of the PCNA technique, which avoids potential perturbations occurring during organ culture, has considerable appeal as a marker for colorectal cancer risk, but additional studies are needed to correlate PCNA with neoplastic risk. PMID- 9332766 TI - Development and validation of an instrument to measure factors related to colorectal cancer screening adherence. AB - This report describes the development and refinement of a set of scales for use in research on predictors of colorectal cancer screening adherence. The study population included 2693 of 4490 eligible white male automotive employees who answered a mailed questionnaire (60% response rate) on beliefs and attitudes related to colorectal cancer and screening. Exploratory and confirmatory factor analyses and multitrait scaling analysis were used to evaluate the construct validity of a priori scales developed to measure salience and coherence, perceived susceptibility, worries about screening, screening efficacy, social influence, and intention. Analyses supported the construct validity of scales for salience and coherence, perceived susceptibility, and worries about screening. Four items originally assigned to the salience and coherence construct loaded on a separate factor that appeared to measure self-efficacy. There was no empirical support for scales measuring screening efficacy and social influence, and there was limited empirical support for a scale measuring intention. Confirmatory factor analysis of the scales measuring salience and coherence, self-efficacy, perceived susceptibility, and worries about screening showed a similar factor structure in white men with and without a personal history of polyps, indicating that the scales may be useful for studies of both colorectal cancer screening and surveillance. Multitrait scaling analysis showed some support for internal consistency reliability of those scales in women (n = 42) and in African-American men (n = 56), and there was some support for the factor structure in those two subgroups. Future studies should evaluate the psychometric properties of these and similar scales in diverse population subgroups. PMID- 9332767 TI - Gas chromatographic-mass spectrometric determination of benzo[a]pyrene and chrysene diol epoxide globin adducts in humans. AB - The efficacy of a newly developed gas chromatography-negative ion chemical ionization-mass spectrometry-selected ion monitoring (GC-NICI-MS-SIM) assay for measuring globin adducts of benzo[a]pyrene (B[a]P) and chrysene diol epoxides in human was evaluated. In this pilot study, smokers and nonsmokers were selected as exposed and nonexposed groups. Using [2H12]r-7,t-8,9,c-10-tetrahydroxy-7,8,9,10 tetrahydrobenzo[a]pyren e ([2H12]trans,anti-B[a]P-tetraol) as an internal standard, B[a]P-tetraols released from globin after hydrolysis and derivatization were quantified by GC-NICI-MS-SIM. Levels of trans-1,2-dihydroxy-3,4-epoxy 1,2,3,4-tetrahydrochrysene (chrysene-DE)-globin adducts were estimated by assuming that the recovery and the MS response of the perdeuterated B[a]P-tetraol internal standard reflected the recovery and MS response of chrysene tetraols. The assay was found to be reproducible and sensitive enough to detect both analytes in all samples. The mean levels of B[a]P-tetraols released from the corresponding benzo[a]pyrene diol epoxide (BPDE) globin adducts in smokers were significantly higher than those in nonsmokers, i.e., 2.6 +/- 0.6 SE fmol/mg globin (ranging from 1.2 to 7.8 fmol/mg globin) in smokers and 0.97 +/- 0.05 SE fmol/mg globin (ranging from 0.7 to 1.3 fmol/mg globin) in nonsmokers (P < 0.01). Interestingly, estimated levels of chrysene-DE-globin adducts in the same subjects were about two orders of magnitude higher than those of the globin adducts of BPDE. The mean of the chrysene-DE adducts in smokers was estimated to be 310 +/- 30 SE fmol/mg globin (ranging from 190 to 460 fmol/mg globin) and that in nonsmokers was 250 +/- 25 SE fmol/mg globin (ranging from 110 to 380 fmol/mg). Although the estimated mean of chrysene-DE adducts with globin in smokers appeared to be about 25% higher than in nonsmokers, the difference was not significant (P = 0.06). The results of this study demonstrate the feasibility of the GC-NICI-MS-SIM method for measurement of BPDE globin adducts in humans. PMID- 9332768 TI - Prognostic value of specific KRAS mutations in lung adenocarcinomas. AB - Adenocarcinomas of the lung remain a significant public health problem. Locally defined (stage I) tumors are considered amenable to resection with curative intent. However, only about 45% of these patients survive for 5 years. The median survival for more advanced tumors is drastically lower. Much research has been focused on identifying a valid genetic biomarker of prognosis. Mutations of the proto-oncogene KRAS have been identified by some groups as being a valid prognostic indicator for adenocarcinoma of the lung. To evaluate the effect of KRAS gene mutation on the survival of patients with lung adenocarcinoma, 181 archival tumors were examined by PCR and denaturing gradient gel electrophoresis. Mutations in either codon 12 or 13 were found in 31.5% of the samples. The most common mutation was a G-->T transversion in codon 12, representing 66.7% of the mutations. No difference was observed in the survival of patients with a KRAS mutation versus those whose tumors contained wild-type KRAS. This lack of difference was also observed when the analysis was restricted to those with stage I tumors or when patients with stage I or II disease were grouped together. However, certain amino acid substitutions, including cysteine, arginine, and aspartate, indicated a significantly poorer prognosis, whereas hydrophobic amino acid substitutions showed a significantly better prognosis than wild-type (P = 0.04). Sample sizes were small for this analysis due to the number of possible mutations. As expected, the stage of tumor at resection was the most significant predictor of outcome. Based on this study of 181 patients from two major medical centers located in different cities, we conclude that KRAS mutation status is not a satisfactory predictor of prognosis in lung adenocarcinoma, but the substitution of a polar or charged amino acid for the wild-type glycine residue may be a negative prognostic indicator. PMID- 9332769 TI - DNA image cytometric measurement as a surrogate end point biomarker in a phase I trial of alpha-difluoromethylornithine for cervical intraepithelial neoplasia. AB - Cervical intraepithelial neoplasia grade 3 (CIN 3) is considered a high-risk precursor of invasive cervical cancer. alpha-Difluoromethylornithine (DFMO) is a promising antiproliferative chemopreventive agent. The purpose of this study was to evaluate image cytometric measurement of nuclear DNA (ICM-DNA) as a surrogate end point biomarker (SEB) in a Phase I trial of DFMO for CIN. Thirty patients with CIN 3 were treated with DFMO at five doses, ranging from 0.0625 to 1.0 g/m2/day, for 1 month. Half of the patients had histological responses. Twenty five pre- and posttreatment cervical biopsy specimens (from 11 responders and 14 nonresponders) were available for this analysis. ICM-DNA was performed on 4 micron sections cut from formalin-fixed tissue blocks and stained with a thionin SO2 Feulgen reaction. ICM-DNAs for each case were expressed as normalized measurements (against the nuclear modal absorbance of lymphocytes) of the absorbance of each cell of interest and were presented in bar histograms. The mean normalized summed absorbance (sigma ODn) was obtained as a mean histogram of the cell population of interest. Nineteen (76%) of 25 patients had a significant decrease in sigma ODn after DFMO treatment. Posttreatment values were significantly lower than pretreatment values in a paired analysis, and responders had significantly lower values than nonresponders. Analyses of different ICM-DNA references, including percentile values of sigma ODn distribution, DNA malignancy grade, and 5c exceeding rate, showed a decrease of mean sigma ODn during DFMO treatment. In addition, the summed posttreatment sigma ODn histograms also showed progressively shorter right shoulders compared with pretreatment histograms in both responders and nonresponders. We concluded that the modulation of sigma ODn reflected the chemoprevention effect of DFMO even before morphological changes appeared, and thus, ICM-DNA may be useful as a SEB in chemoprevention trials of DFMO. Additional reasons for using ICM-DNA as a SEB are the relative simplicity of its use, the high accuracy of the results, the low cost of the reagents, the ability to use small tissue samples, and the objectivity and reproducibility of the procedure. PMID- 9332802 TI - Predictors of destructive periodontal disease incidence and progression in adult and elderly Chinese. AB - This study describes some predictors of new and progressing destructive periodontal disease over a 10-year period in rural Chinese. A total of 398 persons aged 20-80 years, who had participated in a baseline survey of tooth mortality, dental caries and periodontal conditions and were still dentate 10 years later, were reexamined for the same parameters as assessed at baseline. Three different threshold values were used to define new and progressing destructive periodontal disease. Irrespective of the threshold used, most persons experienced new disease. Progressing disease was very prevalent when a 2+ mm disease definition was used, but occurred less frequently at the higher threshold levels. The logistic regression models for 2+ and 3+ mm disease were essentially similar, and showed that women, persons with 104 sites or more, and persons with 0-5% sites with 4+ mm attachment levels had a lower risk of disease progression as well as of new disease than did men, persons with few sites and persons with 6% sites or more with attachment levels 4+ mm. The variables sex, number of sites present, percentage of sites with 4+ mm attachment levels, and presence of mobile teeth were predictive for new disease using a 4+ mm definition. Age, percentage of sites with 4+ mm attachment levels and percentage of sites with 4+ mm pockets were predictive for progressing disease using the 4+ mm disease definition. PMID- 9332770 TI - Correspondence re: G. G. Schwartz, Multiple Myeloma: Clusters, Clues, and Dioxins. Cancer Epidemiol. Biomark. Prev., 6: 49-56, 1997. PMID- 9332803 TI - Predicting which adult patients will need treatment over the next year. AB - This prospective study was conducted to determine factors important in predicting which regularly attending adult patients would receive first, restorations or extractions for any reason (receiving treatment) and, second, restorations or extractions undertaken specifically for caries (receiving treatment related to caries). Baseline and incremental clinical data were obtained from 24 general dental practitioners on a group of their regularly attending, dentate adult patients over a 12-month period. The patients completed a postal questionnaire with questions relating to dental health behaviour, attitudes, knowledge, and social factors. Complete data were obtained from 2553 patients. Thirty-one variables were identified as potential predictors for the two dependent variables receiving treatment and receiving treatment related to caries, and logistic regression models were fitted. Receiving treatment was associated with having fewer sound teeth and more anterior fillings, posterior fillings and crowns (P < 0.001). The dentist's prediction of the need for treatment related to caries and the patient's own prediction of the need for a filling were also important in the model (P < 0.001). Some of these variables, together with having received recent medical treatment and taking sugar in tea or coffee were also found to predict treatment related to caries. The model for receiving treatment related to caries was more successful at predicting the patient's individual risk but the model for receiving treatment was slightly better at classifying patients into whether or not they received treatment. It is reassuring that the common assumptions made by the dental practitioners of their patient's risk have received statistical validation. PMID- 9332804 TI - Control of occlusal caries in the first permanent molars by oral hygiene. AB - Caries of the pits and fissures of permanent teeth continues to be a problem for children, newly erupted permanent molars being particularly at risk. Oral hygiene measures have been shown to be able to reduce the incidence of caries. The aim of this study was to compare the caries-preventive effects on newly erupted first permanent molars of a professional tooth cleaning and oral health education program (test) with a standard preventive program (control), comprising selective fissure sealing and application of topical fluorides. School Dental Service clinics of the Health Department of Western Australia, in Perth, were assigned to four test or four control clinics. Schoolchildren, mean age 6.3 +/- 0.3 (s) years with sound, newly erupted first permanent molars were included in the study (207 test, 197 control). After 12 months, 186 test and 163 control children were examined by an examiner who was 'blind' to the test or control status of the children. Caries of the first permanent molars developed in 34 test and 35 control children; the estimated risk ratio was 0.86 (95% CI 0.56, 1.30). Children in the test group had an average DFT score of 0.26 +/- 0.62 compared with 0.29 +/ 0.64 DFT in the control group (t-test, P = 0.67). The 12-month results suggest that there was no statistically significant difference between the caries preventive effects of a professional tooth cleaning and oral health education program and a program based on selective fissure sealing and application of topical fluorides. PMID- 9332805 TI - Derivation and validation of a short-form oral health impact profile. AB - Growing recognition that quality of life is an important outcome of dental care has created a need for a range of instruments to measure oral health-related quality of life. This study aimed to derive a subset of items from the Oral Health Impact Profile (OHIP-49)-a 49-item questionnaire that measures people's perceptions of the impact of oral conditions on their well-being. Secondary analysis was conducted using data from an epidemiologic study of 1217 people aged 60+ years in South Australia. Internal reliability analysis, factor analysis and regression analysis were undertaken to derive a subset (OHIP-14) questionnaire and its validity was evaluated by assessing associations with sociodemographic and clinical oral status variables. Internal reliability of the OHIP-14 was evaluated using Cronbach's coefficient alpha. Regression analysis yielded an optimal set of 14 questions. The OHIP-14 accounted for 94% of variance in the OHIP-49; had high reliability (alpha = 0.88); contained questions from each of the seven conceptual dimensions of the OHIP-49; and had a good distribution of prevalence for individual questions. OHIP-14 scores and OHIP-49 scores displayed the same pattern of variation among sociodemographic groups of older adults. In a multivariate analysis of dentate people, eight oral status and sociodemographic variables were associated (P < 0.05) with both the OHIP-49 and the OHIP-14. While it will be important to replicate these findings in other populations, the findings suggest that the OHIP-14 has good reliability, validity and precision. PMID- 9332807 TI - Effect evaluation of an oral health education programme in primary schools in Tanzania. AB - This study aimed to assess the clinical oral health outcome effects among schoolchildren participating in a school-based oral health education (OHE) programme. Local social, cultural and environmental conditions were determinants of the school-based OHE programme, which was compiled on the basis of prevailing beliefs and on what teachers and educational authorities considered to be important for the oral health of schoolchildren. Consequently, the practical aspects of oral hygiene and information on the cause and prevention of caries and gingivitis were the components of oral health education. The teachers were prepared to carry out weekly supervised toothbrushing sessions and monthly lessons on aspects of oral health for the school year in grade 4. Eight participating schools were selected for the clinical effect evaluation and four non-participating schools served as the control. In total, 309 children from the participating schools and 122 children from the non-participating schools were available for the evaluation. Their ages varied between 9 and 14 years. The mean plaque score, calculus score and gingival bleeding score at baseline and at follow-up examinations 3, 8, 15 and 36 months later were not significantly different for participating schools and controls. The mean DMFT value at baseline was 0.4 and 3 years later 0.9 in both the participating and control schools. In conclusion, the present study shows that the implemented school-based OHE programme did not result in significant reductions of the clinical parameters measured. PMID- 9332806 TI - Symptoms experienced during periods of actual and supposed water fluoridation. AB - Fluoridation of water is a controversial measure because of the suspicion that it has harmful effects on health. Opinions differ as to the reality of these fears. In Kuopio, after distressing disputes over the fluoridation issue, the City Council decided to stop fluoridation at the end of 1992. In fact, however, it was discontinued at the end of November, one month early, without the public being told. The aim of this study was to find out whether the occurrence of 25 selected symptoms was connected with exposure to fluoridated water. In order to do this we compared the prevalence of symptoms during the months before and after the undisclosed cessation of fluoridation and after the cessation had been officially announced. Postal inquiries concerning symptoms were sent to 1000 randomly selected adults in November, to a further 1000 in December 1992 and again to the same 2000 people in March 1993. The response rates were 40-26%. The percentage of those with two or more symptoms was the same (45%) in November and in December but decreased to 32% in March. The mean number of symptoms per respondent decreased from 1.9 in November to 1.4 in March (P < 0.001) and in December-March from 1.8 to 1.2. The decrease was most significant for symptoms related to the skin. Since the occurrence and mean number of symptoms were fairly similar during actual and supposed fluoridation, the results do not support the theory that the symptoms considered in this study are caused by the physical effect of fluoridated water. On the other hand, the significant reduction in the number of symptoms only after the respondents had become aware of the discontinuation of fluoridation reveals that fluoridation may have psychological effects which present as perceived symptoms. PMID- 9332809 TI - Gender and age differences in attitudes to dental pain and dental control. AB - In the literature, it is usual to find women and younger subjects reporting higher levels of dental anxiety than men and older subjects. Fear of pain was found to be the most important predictor of dental anxiety and issues of control were also related to such anxiety. Therefore, it was predicted that gender and age differences would be reflected in attitudes to pain and control. Subjects were randomly selected from the voters' list in metropolitan Toronto and mailed a questionnaire with a request for cooperation in a study of their thoughts, feelings, and behaviour regarding dental treatment. The questionnaire included demographic data, measures of dental anxiety and painful experiences as well as the Pain Anxiety Symptoms Scale and the Iowa Dental Control Index. The results supported the main predictions. In addition, attitudes to pain and control were found to be complex phenomena with characteristic gender differences. PMID- 9332808 TI - Oral disadvantage among dentate adults. AB - Oral disadvantage can be defined as the avoidance of certain daily activities because of decrements in oral health. These decrements include oral disease and tissue damage, pain, and functional limitation. The Florida Dental Care Study (FDCS) is a longitudinal study of changes in oral health, which included at baseline 873 subjects who had at least 1 tooth, were 45 years old or older, and who participated for an interview and clinical examination. Three objectives of the FDCS are: (1) to describe selected psychometric properties of the measurement of oral disadvantage; (2) to describe oral disadvantage in a diverse sample of dentate adults; and (3) to describe the relationship between disadvantage and other aspects of oral health, such as disease/tissue damage, pain, and functional limitation. The prevalence of oral disadvantage within the previous 6 months, using eight self-reported measures, ranged from 5% to 25%, depending upon the measure. Factor analysis suggested that oral disadvantage is best described as three factors: disadvantage due to (1) oral disease/tissue damage, (2) oral pain, and (3) oral functional limitation. Irregular dental attenders, poor persons, and blacks had the highest prevalence of oral disadvantage. Clinical measures of oral disease/tissue damage, self-reported measures of oral disease/tissue damage, oral pain, and oral functional limitation were strongly associated with the presence of oral disadvantage. In multivariate analyses that accounted for differences in clinical measures of disease/tissue damage, self-reported disease/tissue damage, oral pain, and oral functional limitation, females were more likely to report disadvantage due to disease/tissue damage, and middle-aged persons and irregular dental attenders were more likely to report oral disadvantage due to pain. In these same regressions, differences in disadvantage due to race, poverty status, socioeconomic status, and rural/urban area of residence were not evident. These results have implications regarding the use of oral disadvantage to assess the long-term effectiveness of dental care. PMID- 9332810 TI - Nonresponse bias in a national study of dentists' infection control practices and attitudes related to HIV. AB - The aim was to investigate late response and nonresponse bias in a survey related to HIV and infection control. Questionnaires with ID numbers were mailed to a stratified random sample of dentists in Canada with additional mailings 4 and 7 weeks later (n = 6444). We compared responses received after < 4 weeks, 4-7 weeks, > 7 weeks. Extrapolation was used to estimate nonresponse bias. Univariate analyses showed significant differences between responses received < 4 weeks after initial mailing and those received later for items on sociodemographics, knowledge, infection control practices and attitudes: late responders were more likely to report that they would refuse to treat any patients with HIV (P < 0.01). Multiple logistic regression indicated that the best predictors of responses received > or = 4 weeks were disagreement that HBV is more infectious than HIV (OR = 1.7); unwillingness to attend a dentist who treats HIV/AIDS patients (OR = 1.3); incorrect perception of the risk of HIV infection after an HIV-contaminated needlestick injury (OR = 1.2); and sometimes or never heat sterilizing handpieces after each patient (OR = 1.2). Extrapolation indicated that the percentage of all respondents who reported refusal to treat (15.2%) would have been 17.1% if a 100% response rate had been obtained. We found significant evidence of late response and nonresponse bias primarily in knowledge and fears related to HIV infectivity; however, the impact on the final results was small and we concluded that additional follow-up to improve response rates would not be worthwhile. PMID- 9332811 TI - Dental fluorosis in 12-15-year-old rural children exposed to fluorides from well drinking water in the Hail region of Saudi Arabia. AB - To investigate the relationship between fluoride levels in well drinking water, severity of dental fluorosis and dental caries in the Hail region of Saudi Arabia, 2355 rural children aged 12-15 years were examined. Over 90% of the children had fluorosed teeth and chi-square tests showed a strong association (P < 0.001) between fluoride level (0.5-2.8 ppm) in well drinking water and severity of dental fluorosis. Although regression analysis showed a statistically significant relationship (P < 0.001) between fluoride concentration and caries experience, the amount of variation explained was very low (R2 = 0.9%). Since fluoride in well water had little influence on caries experience and is causing dental fluorosis, it should be removed by defluoridation or the rural population should be provided with an alternative source of drinking water with lower fluoride concentration. PMID- 9332812 TI - An investigation of the disposal of hazardous wastes from New Zealand dental practices. AB - A national survey was conducted to investigate current procedures in New Zealand dental practices for disposal of clinical waste. A questionnaire was sent out to all dental practices in New Zealand, and non-returns were followed up by two further mailings. From three mailings 767 useable questionnaires were returned (71.3% of those sent out, 79.0% of those potentially valid). Responses indicated that 56.4% of dental practices disposed of bloody swabs into the waste paper bin, and 24.4% disposed of contaminated sharp items into the general household refuse collection. Qualitative interviews with dental practitioners revealed a lack of concern about disposal of contaminated waste into the general waste. The existence of legislation governing waste disposal was not sufficient to motivate many practitioners to comply with guidelines. In some areas there was no specialised waste disposal service available, but some dentists had rejected a specialised service on the grounds of cost or inconvenience. Substantial efforts were made to salvage amalgam waste to be sold for scrap. PMID- 9332813 TI - Effectiveness of a dental preventive program on plaque index results in 8-year old children of Bergamo, Italy. AB - The study presents the results of a preventive program based on a children's book and carried out in the elementary schools of Bergamo, Italy. The book was published to improve 8-year-old schoolchildren's knowledge of primary dentition, dental plaque, nutrition, oral hygiene, fluoride and regular dental visits. The study was experimental. A total of 440 schoolchildren were selected to test the effectiveness of the book. They were divided into four groups: two groups underwent the training program and the other two were control groups. Measurements were made using the Quigley-Hein index before and after the program. A significant difference reduction of the plaque index of -0.6 was observed (P < 0.005). PMID- 9332814 TI - Oral hygiene, sucrose consumption and dental caries prevalence in adolescent systemic fluoride non-users. PMID- 9332815 TI - Use of flow cytometry techniques in studying mechanisms of apoptosis in leukemic cells. AB - The use of flow cytometric techniques has significantly aided the rapid advancement of our understanding of the process of apoptosis. Our laboratory is currently involved in investigating the ways in which cells can be induced to die and some of the signals that may be involved. To this end, we are using flow cytometry to detect apoptosis in cell cultures and to measure intracellular changes that occur during apoptosis, in particular, alterations in parameters such as mitochondrial function, oxidative stress, and gene expression. In this review of recent and ongoing research in our laboratory, we outline our studies on mechanisms of apoptosis by cytotoxic drugs, particularly in relation to the involvement of oxidative stress. In addition, we are studying the increased sensitivity to apoptosis seen in K652 cells that have been transfected with the p53 gene. PMID- 9332816 TI - The sensitive detection and quantitation of antibody to HCV by using a microsphere-based immunoassay and flow cytometry. AB - Antibody to HCV core and NS3 was quantified by using a microsphere immunoassay and flow cytometry. Antibody to core and NS3 was elevated in the 85 seropositive blood donors tested. The amount of either antibody varied over two logs although greater variation was seen with the antibody to NS3 than was seen with antibody to core. In three documented acute HCV cases, the microsphere assay detected antibody prior to antibody detection using the reference methods. Twenty donor samples were indeterminate by the reference methods: 45% of these were indeterminate with the microsphere assay while 25% were negative and 30% were positive. As compared to enzyme immunoassay the microsphere assay showed a 5-fold increase in sensitivity. The microsphere assay demonstrated increased sensitivity for the quantification of specific antibody to HCV core and NS3 and was useful in resolving a significant proportion of indeterminate samples. PMID- 9332817 TI - Comparative genomic hybridization in hypotriploid/hyperdiploid tumors. AB - Hypotriploidy/hyperdiploidy ("intermediate ploidy") often occurs in testicular germ cell tumors of adolescents and adults. Disomic and trisomic chromosomes represent significant parts of the tumor genome and a few chromosomes fall outside the two- to three-copy number range. We performed comparative genomic hybridization (CGH) with DNA isolated from a cell line from a case of testicular germ cell tumor of adolescents and adults and found most of the ratio values to be dislocated from the baseline 1.0 and placed adjacent of the diagnostic thresholds of 0.8 and 1.2. We attributed that to the fact that, in current software packages for analysis of CGH, the fluorescence ratio baseline is assumed to correspond to the copy number of most loci of the genome. We then evaluated, instead of the commonly used fluorescent ratio value from the whole metaphase, the use of the fluorescence ratios of single chromosomes. The results permitted a clear distinction between the chromosomes with two and three copies and, in particular, of the regions deleted or amplified outside the two- to three-copy range. We concluded that the evaluation of unbalances of DNA copy number in intermediate ploidy cases is best carried out using multiple normalization. PMID- 9332818 TI - Three-dimensional visualization and quantitation of fibrin in solid tumors by confocal laser scanning microscopy. AB - Fibrin forms part of the stroma essential for growth of solid tumors. Anticoagulants reduce primary tumor growth and tumor metastasis in murine and some human tumors. These effects may be partly mediated by reduction of intra tumor fibrin, although there are no quantitative data to support this hypothesis. We therefore evaluated the effect of warfarin on fibrin deposition in a subcutaneously (s.c.) implanted murine tumor using confocal laser scanning microscopy (CLSM). AJ mice received no treatment (n = 6) or sodium warfarin (3.5 mg/L in drinking water, n = 5). All animals received 2 x 10(6) syngeneic Neuro2a neuroblastoma cells s.c. After 14 days, primary tumors were excised and placed in liquid nitrogen. Warfarin treatment resulted in a small, but significant (P < 0.05), decrease in wet tumor weight. Frozen sections (20 microns) were incubated with goat anti-mouse fibrin(ogen) or normal goat serum (isotypic control) and stained with FITC-conjugated rabbit anti-goat antibody. Using a Multiprobe 2001 CLSM (Molecular Dynamics, Sunnyvale, CA), 20 serial optical sections were taken from five, randomly chosen, high power fields (60x objective) for each slide. A threshold excluded all fluorescence except that from structural components within the tumor stroma (fibrin). The volume of fibrin in each section series was determined, and the percentage of tumor volume occupied by fibrin calculated. Intra- and inter-assay variation were assessed on serial frozen tumor sections from an untreated animal. The percentage fibrin volume was not significantly different among or within experiments, indicating that the procedure was reproducible. In controls, the median (range) volume occupied by fibrin was 8.1% (2.4-22.3%), whereas in anticoagulated animals, this was reduced to 3.7% (0.4 14.0%; P < 0.001). This is the first quantitative demonstration that warfarin reduces fibrin deposition in solid tumors. We conclude that three-dimensional CLSM is useful for the quantitation of tissue antigens and that the technique may have clinical value. PMID- 9332819 TI - Application of automatic thresholding in image analysis scoring of cells in human solid tumors labeled for proliferation markers. AB - We previously reported an image analysis program that uses four investigator defined parameters including two thresholds, i.e., gray-level threshold (GLT) and hue threshold (HT), to determine the number of cells (TC) and the proliferating cell nuclear antigen (PCNA) or bromodeoxyuridine (BrdUrd) labeling indices (LI) in human solid tumors. The present study investigated if the accuracy and reproducibility of image analysis results can be improved by using computer defined GLT and HT. Three investigators evaluated 142 images on 3 days, using visual analysis and four image analysis routines, which used different combinations of computer- and investigator-selected GLT and HT. The data show that image analysis using computer-selected GLT and HT yielded (i) LI of PCNA and BrdUrd that were indistinguishable from visual analysis, (ii) equal (BrdUrd LI) or better (TC and PCNA LI) inter-day reproducibility relative to visual analysis results, and (iii) results that were equally (TC) or more accurate (LI of PCNA and BrdUrd) with higher inter-day reproducibility (TC and LI of PCNA and BrdUrd) than image analysis obtained using investigator-defined thresholds. We conclude that the use of computer-defined GLT and HT improved the accuracy and reproducibility of image analysis results. PMID- 9332820 TI - Classification of organelle trajectories using region-based curve analysis. AB - A method based on analysis of the region of movement and the functioning of the acto-myosin cytoskeleton has been elaborated to quantify and classify patterns of organelle movement in tobacco pollen tubes. The trajectory was dilated to the region of movement, which was then reduced to give a one-pixel-wide skeleton, represented by a graph structure. The longest line in this skeleton was hypothesized to represent the basic track of the organelle along a single actin filament. Quantitative features were derived from the graph structure, direction of movement on the longest skeletal line, and distance between skeletal line and particle. These features corresponded to biological events like the amount of linear movement or the probability of attachment of an organelle to the actin filament. From 81 analyzed organelle trajectories, 17 had completely linear, 17 had completely non-linear, and 47 had alternating linear and non-linear movement. Selected features were employed for classification and ranking of the movement patterns of a representative sample of the population of organelles moving in the cell tip. The presented methods can be applied to any field where analysis and classification of particle motion are intended. PMID- 9332821 TI - Evaluation of fluorochromes and excitation sources for immunofluorescence in water samples. AB - Fluorescent labelling methods for detecting microorganisms in water have limited sensitivity partly due to the natural autofluorescence from environmental particles. The aim of this study was to examine the autofluorescence of water samples to determine the optimal excitation source and fluorescent labels for minimising background autofluorescence and therefore enhancing sensitive detection of Cryptosporidium oocysts. Particles concentrated from water were examined using fluorimetry at a wide range of excitation wavelengths to determine their autofluorescent properties. Two major peaks were identified emitting at 390 to 510 nm and at 640 to 700 nm. Flow cytometry was used to define the optical properties of oocysts immunofluorescently labelled with a range of fluorochromes. Concentrated water samples were analysed using flow cytometry and the number of particles with fluorescence and light scatter properties similar to the fluorescently labelled oocysts recorded. Fluorescein isothiocyanate exited at 488 nm was the most suitable label for oocysts in untreated water with less than 70 particles having optical properties similar to labelled oocysts, detected in 10 litre concentrates. The fluorochromes CY3, phycoerythrin (PE), and tetramethylrhodamine B thioisocyanate (TRITC) excited at 542 nm were the most suitable labels for oocysts in drinking water with less than 40 particles having optical properties similar to labelled oocysts, detected in 100 litre concentrates. PMID- 9332823 TI - Combined flow cytometry and immunohistochemistry analyses for the assessment of nasopharyngeal carcinoma cell kinetics by in vivo bromodeoxyuridine infusion. AB - Thirty-eight previously untreated patients with a histologically proved diagnosis of nasopharyngeal carcinoma were evaluated for in vivo cell kinetics before treatment by conventional radiation therapy. Thirty-seven tumors were analyzed by flow cytometry. Values of median potential doubling time (Tpot), labelling index (LI), and duration of S phase (Ts) were, respectively, 10.9 days, 3.8%, and 10.8 hours. In 35 cases, the results obtained from two biopsies of the same tumor were compared. A good reproducibility was obtained for LI and Tpot (P < 0.01). Thirty one tumors were analysed by immunohistochemistry and labelling index (HLI) was determined in 24 tumors with a percentage of labelled cells varying from 6.1% to 39.2% (median value = 18.5%). No correlation was found between LI and HLI, but when observations were focused on the restricted group of DNA aneuploid samples, mean values of LI and HLI were closer (respectively, 12.8 +/- 4.5% and 18.3 +/- 7.7%) and a good correlation was obtained (P = 0.01). Moreover, no difference in proliferation was found between diploid and aneuploid tumors. Considering these results, a combined Tpot was calculated that allowed classification of tumors as highly or slowly proliferative. PMID- 9332824 TI - Laser scanning cytometry allows detection of cell death with morphological features of apoptosis in cells stained with PI. AB - Laser scanning cytometry (LSC), a newly developed technology, allowed simple detection of dead cells with morphological features of apoptosis for cells stained with only propidium iodide (PI). HeLa cells were treated with Adriamycin (ADM, 0.5 or 1.0 microgram/ml). A PI fluorescence value (representing DNA content) versus PI fluorescence peak (representing chromatin condensation) cytogram of LSC made it possible to segregate cells with high PI fluorescence peak from others in a cell population and concomitantly to analyze the relationship between the cells and the cell cycle. A fraction of the cells manifesting hypercondensation of chromatin was exclusively present in a cell population treated with ADM. Visual inspection of the cells defined in the cytogram revealed morphological features of apoptosis. LSC analysis facilitates monitoring effects of anticancer drugs on a cell population, because nuclear DNA staining with PI is simple and rapid. PMID- 9332822 TI - Identification and analysis of macrophage-derived foam cells from human atherosclerotic lesions by using a "mock" FL3 channel in flow cytometry. AB - Macrophages are one of the major cell types in atherosclerotic lesions. They are believed to play an important role in the pathogenesis and development of the lesion, but their functional state and phenotypic characteristics are not well understood. Using flow cytometry, we analyzed surface markers of macrophages extracted from tissue digests. However, conventional techniques were hampered by the abundance of cell debris and extracellular lipids, which co-localized with double-positive cells in all fluorescent plots. We therefore developed a method to overcome this problem by using a novel gating technique in multiparameter flow cytometry. This method utilized the third fluorescence channel (FL3) as a "mock" channel, since no antibody conjugated to an FL3-specific fluorochrome was added to the samples. Cells single-positive for macrophage-specific monoclonal antibodies (mAb) conjugated to phycoerythrin (PE) (FL2) were separated from non specific fluorescent particles in the FL2 versus FL3 fluorescent plot and a region excluding debris could be set. This was then used as a gate to exclude debris also in the first fluorescence channel (FL1) vs. FL2 plot in which expression of a panel of activation markers identified by fluorescein isothiocyanate (FITC)-conjugated mAb was analyzed. Using this strategy, we were able to identify and analyze the phenotype of macrophages from human atherosclerotic lesions. We were also able to sort these cells and in this way obtained a preparation of macrophage-derived foam cells from the tissue with little contamination of debris. PMID- 9332825 TI - Four color immunofluorescence detection using two 488-nm lasers on a Becton Dickinson FACS Vantage flow cytometer. AB - Multiparameter flow cytometric analysis is sometimes limited by the availability of directly conjugated monoclonal antibodies or streptavidin-conjugated secondary reagents. While many manufacturers offer a wide variety of monoclonal antibodies or streptavidin reagents directly conjugated to 488-nm excitable fluorochromes, there are not many available that are directly conjugated to 360-nm (UV) or 630 nm (HeNe) excitable fluorochromes. For this reason we attempted to develop a four color immunofluorescence staining protocol on a FACS Vantage using four 488-nm excitable fluorochromes. The fixed configuration of the FACS Vantage limits the feasibility of using four 488-nm excitable fluorochrome simultaneously because of the five fluorescence detectors, two are always electronically delayed. This means that only three signals-FL1, FL2, and FL3-can be detected off the primary 488-nm laser beam. FL4 and FL5 are always delayed, only detecting signals off the second laser beam. Our instrument is configured with an ILT air cooled 488-nm laser in the second position that is used in order to conserve the Coherent Enterprise laser when using only 488-nm excitable fluorochromes. Because of this, we were able to develop a four-color immunofluorescence staining protocol using only 488-nm excitable fluorochromes. PMID- 9332826 TI - Novel flow cytometric method for quantifying nuclear binding of the transcription factor nuclear factor kappa B in unseparated human monocytes and polymorphonuclear cells. AB - The transcription factor nuclear factor kappa B (NF kappa B) regulates the production of a number of pro-inflammatory mediators involved in polymorphonuclear and mononuclear cell activation. Measurement of NF kappa B DNA binding is used as an estimate of cellular activation and is usually measured by the DNA mobility shift assay. Results from the mobility shift assay can be difficult to quantify and do not allow discrimination of activity in different populations of cells in whole blood without prior separation. This paper describes a new flow cytometric method which allows rapid detection and quantification of DNA-bound NF kappa B in white cell populations of whole blood. The technique is sensitive and allows discriminate analysis and quantification of bound NF kappa B in nuclei from polymorphonuclear and mononuclear cell populations in whole blood. PMID- 9332827 TI - The herpesvirus protease: mechanistic studies and discovery of inhibitors of the human cytomegalovirus protease. AB - The herpesvirus protease is a recently identified enzyme which is essential for viral replication. It is found in all herpesviruses and offers a new molecular target for therapeutic intervention. Its genomic structure has recently been described and consists of a large open reading frame which encodes a fusion protein containing an amino-terminal protease domain in-frame with a carboxyl terminal "assembly protein-like" domain. Auto-processing releases the amino terminal protease as a maturational enzyme. The herpesvirus protease has been characterized as a novel serine protease. Four surface accessible sulfhydryl groups have been identified in the human cytomegalovirus (HCMV) protease. Utilizing a fluorogenic DABCYL-EDANS substrate assay, directed screening has identified a class of sulfhydryl-modifying benzimidazolylmethyl sulfoxides which inhibits recombinant HCMV protease. Site-directed mutagenesis studies suggest oxidative modification of surface-accessible HCMV protease Cys138 (and possibly Cys161) by this class of inhibitors. The benzimidazolylmethyl sulfoxide 1 inhibits HCMV protease (IC50 = 1.9 microM), exhibits selectivity vs. mammalian serine proteases, and exhibits antiviral activity in an HCMV infected cell culture assay. PMID- 9332828 TI - Discovery and analysis of inhibitors of the human immunodeficiency integrase. AB - An essential step in the replication of retroviruses is the integration of a DNA copy of the viral genome into the genome of the host cell. Integration encompasses a series of ordered endonucleolytic and DNA strand transfer reactions catalyzed by the viral enzyme, integrase. The requirement for integrase activity in the propagation of HIV-1 in cell culture defines the enzyme as a potential target for chemotherapeutic intervention. We have therefore developed a non radioisotopic microtiter plate assay which can be used to identify novel inhibitors of integrase from random chemical screens and for the bioassay driven isolation of inhibitors from natural products. This assay uncouples various steps in the reaction pathway and therefore can be exploited to characterize inhibitors. In this monograph we describe a series of modifications to the method which facilitate such mechanistic studies using as an example a series of previously described integrase inhibitors. PMID- 9332830 TI - Assemblin, an essential herpesvirus proteinase. AB - The herpesvirus maturational proteinase, called assemblin, is essential for the production of infectious virus and represents a new molecular target for the development of antivirals. A brief summary of the synthesis, structure, and function of this fascinating enzyme is presented here. PMID- 9332829 TI - Antiviral properties of 3-quinolinecarboxamides: a series of novel non-nucleoside antiherpetic agents. AB - Novel antiherpetic 3-quinolinecarboxamides were discovered as part of a drug discovery program at Sterling Winthrop Inc. A major goal of this research was to identify novel non-nucleoside agents possessing activity against acyclovir resistant herpes simplex virus. From screening compound libraries in an HSV-2 plaque reduction assay, 1-ethyl-1,4-dihydro-4-oxo-7-(4-pyridinyl)-3 quinolinecarboxamide (1) emerged as an attractive lead structure. By modifying the quinoline ring at the 1-, 2-, 3-, 4-, and 7-positions, analogues were identified that have up to 5-fold increased in vitro potency relative to acyclovir. In a single dose mouse model of infection the 1-(4-FC6H4) analogue 17, one of the most potent derivatives in vitro, displayed comparable oral antiherpetic efficacy to acyclovir at 1/16 the dose; in a multiple dose regimen, however, it was 2-fold less potent. Mechanism of action studies indicate that these new compounds interact with a different, as yet undefined, molecular target than acyclovir. PMID- 9332831 TI - 2,2'-Dithiobisbenzamides and 2-benzisothiazolones, two new classes of antiretroviral agents: SAR and mechanistic considerations. AB - Substituted 2,2'-dithiobisbenzamides and 2-benzisothiazolones were prepared and shown to possess low microM activity with high therapeutic indices against HIV-1, HIV-2 and SIV in cell culture. The mechanism of antiviral action was determined to be directed toward the nucleocapsid protein (NCp7), which contains two zinc fingers and plays vital roles in the viral life cycle. The "active sulfides" of this study cause the extrusion of zinc from these zinc fingers. Structure activity relationships of the 2,2'-dithiobisbenzamides reveal that the disulfide bond and the ortho benzamide functional groups are essential for activity, with the best compounds having a carboxylic acid, carboxamide, or sulfonamide substituent. The 2-benzisothiazolones are formed from the disulfides both chemically and in vivo and their SAR mimics that of the 2,2'-dithiobisbenzamides. The antiviral activity of the disulfides may require cyclization to the isothiazolones. Two agents, PD 159206 and PD 161374, which showed good antiviral activity, physical properties, and excellent pharmacokinetics in mice, were selected for advanced studies. PMID- 9332883 TI - Intramuscular fosphenytoin (Cerebyx) in patients requiring a loading dose of phenytoin. AB - Fosphenytoin (Cerebyx), is a water soluble prodrug that is rapidly and completely converted to phenytoin. This study reports the injection-site tolerance and safety of intramuscular fosphenytoin (> 10 mg/kg doses) in 60 patients requiring a phenytoin loading dose. Patients received injections at single or multiple sites with volumes ranging from 4 to 30 ml per injection site. The majority of patients had no irritation (erythema, swelling, tenderness, bruising) or complaints of discomfort related to fosphenytoin injection either after injection (95%) or at follow-up (88%). Irritation, when reported, was mild in all cases. Forty of 60 patients (67%) reported transient side effects, primarily involving the central nervous system, such as nystagmus, dizziness or ataxia, which are commonly associated with phenytoin therapy. All patients received prescribed doses; no patient had an injection(s) stopped due to intolerance or side effects. No serious adverse events occurred with intramuscular fosphenytoin. In this study, intramuscular fosphenytoin was demonstrated to be a safe and well tolerated, and in many instances, a preferable alternative to other means of phenytoin loading. PMID- 9332882 TI - Topiramate enhances GABA-mediated chloride flux and GABA-evoked chloride currents in murine brain neurons and increases seizure threshold. AB - The anticonvulsant topiramate is effective in laboratory animals against maximal electroshock seizures, amygdala kindling, and spike-wave discharges and has demonstrated efficacy in humans for the treatment of complex partial seizures. However, its mechanism of action has yet to be clearly elucidated. When the chloride-sensitive fluorescent probe N-(ethoxycarbonylmethyl)-6 methoxyquinolinium bromide (MQAE) was used as a tool for estimating the effect of anticonvulsant drugs on GABA receptor function, topiramate was observed to enhance GABA-stimulated chloride (Cl-) flux. At a therapeutic concentration, topiramate (10 microM) enhanced GABA-stimulated (10 microM) Cl- influx into cerebellar granule neurons but did not significantly increase Cl- influx alone. Phenytoin (10 microM) and acetazolamide (300 microM) did not enhance GABA stimulated Cl- influx. In patch-clamp electrophysiological studies, topiramate also enhanced GABA-evoked whole cell Cl- currents in mouse cerebral cortical neurons in culture. In vivo anticonvulsant studies confirmed that topiramate, like phenytoin, is primarily effective against tonic extension seizures induced by maximal electroshock and is ineffective against clonic seizures induced by the subcutaneously administered chemoconvulsants pentylenetetrazol (PTZ), bicuculline (Bic), and picrotoxin (Pic). In contrast to phenytoin, topiramate, at a dose equivalent to the MES median effective dose (ED50), was found to elevate seizure threshold as estimated by the intravenous PTZ seizure threshold test. Taken together these results support the conclusion that enhancement of GABA-mediated Cl- flux may represent one mechanism that contributes to the anticonvulsant activity of topiramate. PMID- 9332884 TI - Dynorphin and the kappa 1 ligand [3H]U69,593 binding in the human epileptogenic hippocampus. AB - The distribution of dynorphin (DYN), one of its binding sites (kappa 1 receptor) and their relationship to neuronal loss and granule cell hyperexcitability was examined in hippocampi from patients with temporal lobe epilepsy (TLE). In hippocampi that were not the seizure focus (mass associated temporal lobe epilepsy, MaTLE; and paradoxical temporal lobe epilepsy, PTLE) DYN-like immunoreactivity was localized in the dentate granule cells and their mossy fiber terminals within the hilus and area CA3. In hippocampi that were the seizure focus (MTLE), 89% showed an additional band of immunoreactivity confined to the inner molecular layer (IML) of the dentate gyrus, representing recurrent mossy fiber collaterals. In 11% of MTLE patients no staining was found in the IML (MTLE/DYN-). The MTLE/DYN- hippocampi were also characterized by a significantly lower degree of cell loss than in MTLE hippocampi in the dentate granule cell layer, the hilus and CA3. Both MTLE and MTLE/DYN- hippocampi showed evoked epileptiform bursting in granule cells while MTLE showed greater polysynaptic EPSPs and spontaneous excitatory activity. Thus granule cell recurrent collateral sprouting may account for only some aspects of hyperexcitability. In 30% of the MTLE group, hilar neurons of a variety of morphological types expressed DYN immunoreactivity in their somata and dendrites. The density of [3H]U69,593 binding sites in MaTLE and PTLE patients was highest in areas CA1 and the subiculum-regions having little or no DYN-staining. In the dentate molecular layer, hilus and CA3--regions with the most DYN immunoreactivity--there was a low density of ligand binding. The significance of this transmitter/receptor mismatch is yet unknown. PMID- 9332885 TI - Effect of lamotrigine on carbamazepine epoxide/carbamazepine serum concentration ratios in adult patients with epilepsy. AB - Although lamotrigine (LTG) appears to have a low propensity to cause pharmacokinetic interactions with other medications, it has been suggested that LTG may interfere with the elimination of carbamazepine 10,11-epoxide (CBZE), the active metabolite of carbamazepine (CBZ). Evidence for this pharmacokinetic interaction is inconclusive and conflicting, however. We evaluated CBZ apparent oral clearance and the steady-state CBZE/CBZ serum concentration ratios in nine patients (30.8 +/- 7.7 years) with epilepsy prior to and following the initiation of adjunctive treatment with LTG. Overall, CBZ oral clearance was unchanged following the introduction of LTG (5.58 +/- 1.60 vs. 5.81 +/- 1.74 1/h, P = 0.630). Likewise, CBZE to CBZ serum concentration ratios were not significantly different (0.241 +/- 0.082 vs. 0.232 +/- 0.082, P = 0.782). These observations suggest that the addition of LTG did not result in a significant pharmacokinetic interaction involving either CBZ or CBZE. PMID- 9332886 TI - Aberrant expression of GABAA receptor subunits in the tottering mouse: an animal model for absence seizures. AB - The single-locus mutant mouse tottering (tg) is an established model for absence seizures. We have previously reported an impairment in GABA-induced chloride uptake in tg brain [Tehrani and Barnes, Epilepsy Res. 1995;22:13-21]. In order to determine if this alteration in GABAA receptor function can be related to specific receptor isoforms, we examined the radioligand binding properties of GABAA receptors and the expression of GABAA receptor subunit mRNAs in the cerebral cortex. Saturation binding of [3H]flunitrazepam revealed a significantly lower Kd value in tg cortical tissues (1.77 +/- 0.05 nM) in comparison to that for the background C57BL/6J strain (3.23 +/- 0.23 nM), while the Bmax values were indistinguishable. Biphasic displacement of [3H]flunitrazepam binding by 2 oxoquazepam showed that low affinity binding sites account for 36 +/- 7.6 and 51 +/- 7.5% of the total in control and tg, respectively. The level of [35S]-t butylbicyclophosphorothionate (TBPS) binding to tg cortical membranes was 73.6 +/ 5.8% of that in controls. Paired measurements by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed no significant differences in the levels of GABAA receptor alpha 1, alpha 3, alpha 5, beta 2, beta 3, gamma 2 or gamma 3 subunit mRNAs between tg and control cortex. However, tg tissues showed elevated levels of alpha 2- and beta 1-subunit mRNAs, representing 256 and 177%, respectively, those of controls. For the tg cortex, the enhanced expression of GABAA receptor alpha 2 and beta 1 subunits correlates with recombinant subtypes known to have low affinity for 2-oxoquazepam and impaired binding of TBPS. These aberrant properties of GABAA receptors could influence the development or propagation of phenotypic seizures in the tottering mouse. PMID- 9332887 TI - Synaptic vesicle size and shape profile in the kindling model of epileptogenesis. AB - Excitatory synapses were studied in the dentate gyrus ipsilateral to stimulated entorhinal cortex in fully kindled rats 2 weeks after the last (3rd) stage 5 seizure. In previous studies of the same model marked redistribution of synaptic vesicles to a 'strategic position' in the vicinity of the synaptic cleft and a significant enlargement of postsynaptic element were described. In this study the size and shape of synaptic vesicles were evaluated in three zones parallel to the presynaptic membrane of the synaptic cleft. The first zone (0.1 micron) directly adjoined the active zone of presynaptic membrane, the second was from 0.1 to 0.2 micron and the third from 0.2 to 0.3 micron from the presynaptic membrane. The vesicles of both the experimental and control animals were significantly smaller in zone I than in zone II and III and, in the control animals there was a tendency of vesicles to diminish towards the active zone. Such a tendency was not observed in the experimental animals, the largest vesicles were in the middle of zone II and were significantly more rounded in comparison to the vesicles in the controls. On the contrary, the vesicles in zone I and III were more elongated in the experimental animals than in controls. Our results may be taken as a sign of active reconstruction of synaptic apparatus in relation to increased functional readiness of the synapses. Possible mechanisms as well as significance for the kindling and epileptogenesis are discussed. PMID- 9332888 TI - Thalamic glucose metabolism in temporal lobe epilepsy measured with 18F-FDG positron emission tomography (PET). AB - Thalamic glucose metabolism has been studied in 24 patients suffering from temporal lobe epilepsy (TLE) using interictal 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET). A total of 17 patients had a unilateral TL seizure onset, 11 of these patients had a mesial temporal lobe epilepsy syndrome (MTLE), with mesial gliosis and a mesial TL seizure origin. Three patients had a lateral TL seizure origin, and 3 patients had mesial TL tumors. Bilateral TLE was assumed in 7 patients. Only in the patient group with MTLE (n = 11), the ipsilateral thalamic glucose uptake showed a statistically significant lower value when compared to the thalamus of the contralateral side (Wilcoxon paired sign test, P = 0.012). There was a more pronounced hypometabolism in right TLE compared to left TLE. A 'hypersynchronous seizure onset pattern' in ictal EEG was only seen in 6 (26%) patients (1 patient with bilateral, 5 with unilateral TLE). No correlation existed between the thalamic, temporal glucose metabolism and the 'hypersynchronous seizure onset pattern'. PMID- 9332889 TI - Increased excitatory amino acid levels in brain cysts of epileptic patients. AB - We studied two epileptic patients with arachnoid brain cysts by proton magnetic resonance spectroscopy (1H MRS). In addition, histochemical analyses of surgical specimens, cerebrospinal fluid, and cystic fluid were performed in one of the patients. In both patients, greatly increased levels of excitatory amino acids (EAAs) glutamate and aspartate were present in the cystic fluid, while there was only a moderate increase of glutamate in the epileptogenic brain tissue adjacent to the cyst in one of the patients. In non epileptic brain regions, no elevations of the EAAs were present. Since EAAs are involved in induction and maintenance of epileptogenesis, their extremely high concentrations in the cystic fluid may explain seizures in some patients with such brain cysts. Our findings may have therapeutical consequences for patients with drug resistant epilepsy, in whom elevated concentrations of EAAs in the cysts can be verified. Surgery with the aim to create a communication between the cyst and the subarachnoidal space may prevent an accumulation of the EAAs and thus result in a relief of seizures. PMID- 9332890 TI - Diagnostically challenging lesions in head and neck pathology. AB - There are a variety of diagnostically challenging lesions in the head and neck region. Contact ulcer usually occurs within specific clinical parameters (vocal abuse, post-intubation and gastro-esophageal reflux), which should be documented in correlation with the granulation tissue-like response affecting the posterior vocal cords. Spindle squamous cell carcinoma (carcinosarcoma) presents a variably cellular spindle cell proliferation, often with surface epithelial ulceration. The clinical presentation of a firm, polypoid mass in the larynx, combined with the histomorphologic features of a spindle cell tumor, can be confirmed to be of epithelial origin when a portion of the overlying epithelium is seen to blend with the spindle cell component, or when ancillary studies authenticate the epithelial origin of the tumor. The diagnosis of a verrucous squamous cell carcinoma can only be made accurately with an accurate clinical history. The very well differentiated histologic appearance, a broad pushing border of infiltration, a bland epithelial proliferation with scant mitotic activity and "church-spire"-type keratosis coupled with the clinical presentation of a large, locally destructive lesion, can confirm the diagnosis of verrucous carcinoma. A wide variety of disorders can result in midline destructive disease clinically, but a specific etiology must be sought to provide appropriate clinical management. Angiocentric T/NK-cell lymphoma of the sinonasal tract is one such disease. The atypical lymphoid cells are usually angiocentric and angiodestructive in their growth pattern. Identification of the atypical cells in the early stages of disease may be difficult, often requiring multiple biopsies over time with the application of immunohistochemical stains or molecular studies to accurately identify the nature of the infiltrate. Cystic squamous cell carcinoma in the neck is almost always a manifestation of metastatic tumor and not a brachiogenic carcinoma. When specific histomorphologic features are noted (a large, unfilled cyst lined by a ribbon-like or endophytic growth of a "transitional"-appearing squamous epithelium with a limited degree of anaplasia), most of these tumors demonstrate primaries in Waldeyer's ring, often of a very small size. Adequate clinical work-up (pan-endoscopy, extensive radiographic imaging and random biopsies or prophylactic tonsillectomy) is mandatory in order to limit the radiation-therapy ports and to document the location of the primary, yielding an excellent long-term prognosis. PMID- 9332891 TI - Local effects of nitric oxide on vestibular blood flow in the Mongolian gerbil. AB - There is a paucity of studies regarding the regulation of vestibular blood flow (VBF), despite the possibility that vascular alterations may contribute to specific vestibulopathies. The current experiments used the Mongolian gerbil as an animal model since it provides easy surgical access to the vestibular end organs and has been previously used for physiologic studies involving inner ear function. VBF changes were measured in the posterior semicircular canal using laser Doppler flowmetry following round window membrane (RWM) application of the nitric oxide donor 1, 3-propanediamine-N-[4-1-(3-aminopropyl)-2-hydroxy-2 nitrosohydrazi no] butyl (spermine NONOate; SPNO) as a vasodilator. The specificity of the responses induced was tested via pretreatment with an NO scavenger, 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazonline-1-oxyl-3-oxide (carboxy-PTIO; cPTIO). cPTIO, SPNO, vehicle (control) or cPTIO/SPNO were applied to the RWM, during which blood pressure and VBF were monitored for baseline, treatment, and recovery conditions. Results showed concentration-dependent increases in flow, probably resulting from NO's vasodilatory action on local vasculature. cPTIO pretreatment was found to attenuate SPNO-induced VBF increases. These findings support a role of NO in maintaining the vestibular microcirculation. PMID- 9332892 TI - Effectiveness of ribosomal fractions of Klebsiella pneumoniae, Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and the membrane fraction of Kp (Ribomunyl) in the prevention of clinical recurrences of infectious rhinitis. Results of a multicenter double-blind placebo-controlled study. AB - A multicenter, double-blind, placebo-controlled study was conducted to investigate the efficacy of an immunostimulant, Ribomunyl, in the prevention of recurrences of infectious rhinitis in adults. This trial involved 327 patients (168 Ribomunyl treated and 159 placebo cases) with an average of 4.3 +/- 1.8 rhinitis episodes per patient recorded during the year preceding the study. The main criterion of efficacy was the cumulative number of recurrences of infectious rhinitis during a 6-month follow-up period, as analyzed by standard tests. An additional analysis of relative risk of recurrences used multivariate failure for time data. Ribomunyl was effective throughout the study period, starting from the first month of treatment: a mean of 1.0 +/- 1.1 recurrences was recorded in the Ribomunyl group as compared to 1.5 +/- 1.4 recurrences in the placebo group; this indicated one-third fewer infections (P = 0.001). The protective effect of Ribomunyl on the relative risk for recurrences was estimated to be 0.58 by multivariate analysis (95% CI: 0.43-0.78, P = 0.0001). Analysis of secondary criteria also favored Ribomunyl: 38.5% less antibiotic courses per patient (0.8 +/- 1.3 vs 1.3 +/- 1.6; P = 0.002) and the number of days with antibiotics (5.6 +/- 9.3 vs 9.1 +/- 12.1; P = 0.002). PMID- 9332893 TI - Cytokeratin and vimentin expression in normal epithelium and squamous cell carcinomas of the larynx. AB - The immunohistochemical expression patterns of cytokeratin polypeptides and vimentin were investigated in normal epithelia and squamous cell carcinomas of the larynx with special emphasis on tumor grading. During malignant transformation of epithelial cells, the cytokeratin expression patterns changed, depending on the differentiation grade of the carcinomas. In low-grade carcinomas, the expression patterns were close to those of the normal epithelium. With increasing tumor grade, there was decreased expression of stratification cytokeratins and increased expression of basal cell, simple cell and hyperproliferation-related cytokeratins. Increasing tumor grade was also associated with the expression of vimentin, a cytoskeletal protein of mesenchymal cells. No relationship was found between vimentin expression and the presence of lymph-node metastases. PMID- 9332894 TI - Results of bone conduction following surgery for chronic ear disease. AB - Preoperative and postoperative bone conduction thresholds were compared in 181 chronic ears operated on over a 5-year period between 1990 to 1994. In the majority (92%) of cases the bone conduction thresholds remained unchanged (+/-10 dB). Nine ears (5%) showed better thresholds after surgery, with improvements ranging from 11 dB to 25 dB. This improvement was especially noted in ears with severe tympanic pathology. One ear with a large labyrinthine fistula became totally deaf after surgery. In 5 ears (3%) bone-conduction thresholds deteriorated, but remained measurable at all frequencies tested. In these latter cases this impairment ranged from 11 dB to 27 dB. Cholesteatomatous ears having intact ossicular chains were found to be at the highest risk of inner ear damage when "canal wall-down" mastoidectomies were performed. Methods for prevention of sensorineural hearing loss following chronic ear surgery are discussed. PMID- 9332895 TI - A prospective study using rhinomanometry and patient clinical satisfaction to determine if objective measurements of nasal airway resistance can improve the quality of septoplasty. AB - In the ENT Department of University Central Hospital, Turku, the waiting list for elective septoplasty grew to 4 to 5 years in the late 1980s. Therefore, a prospective clinical project was initiated during which all patients waiting for septal surgery were re-examined and nasal airway function was measured with rhinomanometry. Patients with high nasal resistance or other specific indications for nasal obstruction were selected for surgery (n = 432). The remaining patients were excluded from surgery and followed up (n = 284). Results showed that if patients were referred for septal surgery without rhinomanometric study, about 10% became symptom-free within 3 to 5 years. Patients operated on after defining a high preoperative intranasal resistance had a higher postoperative satisfaction level (85%) than those operated on with normal nasal resistances but other indications for correcting the nasal septum (69%). After 3 years, the majority of patients not treated surgically were satisfied with their conservative treatments, although certain patients still required some form of nasal surgery to relieve recurring nasal and/or sinus complaints. PMID- 9332896 TI - Effects of botulinum toxin on vocal tract steadiness in patients with spasmodic dysphonia. AB - Since laryngeal botulinum toxin (BTX) injections have become the treatment of choice for spasmodic dysphonia, the purpose of this study was to examine its effects on the stability of the upper vocal tract as compared to the effects on glottic stability. Two different acoustic methods were used to analyze voice samples from 16 patients with adductor-type spasmodic dysphonias before and after BTX therapy and from a normal control group. Independent acoustic analyses were used to determine laryngeal and upper vocal tract stability. The results showed significantly higher values for the standard deviation of fundamental frequency (SDF0), reflecting laryngeal instability, for the patient group than for the control group and an impressive improvement for the patients after BTX therapy. Further, the equally high values of SDF0 for the initial second and a second from the midsegment of phonation were differentially reduced by BTX therapy, resulting in a normal pattern of laryngeal stability during sustained phonation. The variability of the first and second formants, reflecting upper vocal tract instability, showed higher values for the patients compared with the control group, but this difference was not statistically significant. The present findings showed that BTX injections to the thyroarytenoid muscle had no discernible effect on stability of the upper vocal tract. PMID- 9332897 TI - Electron microscopic localization of nitric oxide I synthase in the organ of Corti of the guinea pig. AB - Nitric oxide synthase (NOS) activity has been detected previously in the mammalian cochlea at a light microscopic level. Here we present results of electron microscopic analysis for post-embedding immunoreactivity of neural-type NOS I in the cochlea of the guinea pig. Strong enzyme immunoreactivity was identified in the cytoplasm of inner and outer hair cells. Gold-labeled NOS I antibodies were mainly located in electron-dense areas of the cytoplasm, whereas electron-lucent regions of the receptor cells were nearly free from any immunoreactivity. In both types of hair cells anti-NOS I antibodies were also visible in the cuticular plates, hair bundles and nuclei. Further ultrastructural analysis revealed that the submembranous cisternae of the outer hair cells were nearly free from any reaction product, demonstrating that the whole cytoplasm of this hair cell was not immunoreactive. Other NOS I immunoreactivity was identified in the cuticular plates of the inner and outer pillar cells and in the cytoskeletal elements located in the apical parts of Deiter cells, forming the lamina reticularis or in cytoskeletal-containing regions in basal Deiter cells. Anti-NOS antibodies were visible in the nuclei of various cell types. Our findings suggest that nitric oxide produced by NO I synthase in the organ of Corti may act as a modulator of hair cell physiology during the processes of signal transduction with frequency selectivity. PMID- 9332898 TI - Aspiration and speech shunt stenosis in near-total laryngectomy patients. AB - A near-total laryngectomy or near-total laryngopharyngectomy with the creation of a speech shunt was carried out on 66 laryngopharyngeal cancer patients. Intelligible shunt speech was obtained in 50 patients (76%), while stenosis of the shunt occurred in 13 patients (20%), and asymptomatic aspiration in 4 (6%). Poor voice production was usually due to shunt stenosis, a fibrotic band surrounding the shunt, or local recurrence of tumor. Perioperative wound infection and postoperative irradiation did not interfere with the development of shunt speech. In 12 patients with shunt stenosis, revision surgeries were performed to augment the shunt openings. During these procedures, the stenotic shunt was opened and a silastic tube was introduced into the shunt lumen to serve as a stent. For the moderately to severely stenotic shunt, the mucosal defect present was covered with a free skin graft or a sternocleidomastoid myoperiosteal flap. Following treatment, results demonstrated that 10 of the 12 patients acquired satisfactory shunt speech. In two unsuccessful cases, the causes of failure were wound infection and graft necrosis and were presumed to be complications of previous irradiation. PMID- 9332899 TI - Late-onset schizophrenia and the delusional disorders in old age. PMID- 9332900 TI - The epidemiology of functional psychoses of late onset. AB - For the functional psychoses of late life, epidemiological information comes from two sources: studies of persons who have reached psychiatric services; and surveys of elderly persons sampled from the general population. A conspectus of published data from both sources leads to the following conclusions: States phenomenologically similar to those found in clinics do occur in the community in non-trivial numbers. There is no notable divergence in the information obtained from clinical series and from population-based surveys. These states are more common in women, they become more common with increasing age and are sometimes associated with decline in cognitive performance or with degenerative changes in the brain revealed by neuroimaging. Genetic factors appear to be less important than in early-onset psychoses but remain ill-defined, and the roles of social isolation and disorders of personality have not yet been sufficiently elucidated. Both clinical and community-based studies have found an association with sensory impairment. The community-based data suggest that paranoid symptoms may be detectable at subclinical level, and an association between them and cognitive impairment is demonstrable in individuals who are not diagnosable cases either of psychosis or of dementia. Differences exist between late-onset paranoid psychoses and affective psychoses in symptomatology and response to treatment. These observations confirm the importance of the late-onset psychoses for research directed towards uncovering the origins of psychotic symptoms in any age group. PMID- 9332901 TI - Neuropathological and neuroradiological correlates of paranoid symptoms in organic mental disease. AB - This paper reviews paranoid symptoms in older patients with organic mental disease. We have taken a dual approach to this topic, examining patients with dementia in whom paranoid symptoms are present and also assessing the presence of organic brain changes in patients diagnosed as having late-onset schizophrenia, paraphrenia or delusional disorder. (For the sake of continuity and not wishing to pre-empt any discussion of the nosological categorisation of late-onset psychoses, we refer to late-onset persecutory state as paraphrenia.) Firstly, there is a description of the various paranoid symptoms which have been described in patients with dementia. Secondly, brain imaging studies are discussed which have highlighted changes in patients with paraphrenia and particular associations between psychotic phenomenology and brain changes in patients with dementia. Thirdly, neuropathological and neurochemical changes in the brains of patients with dementia in whom paranoid symptoms have been present are presented. We intersperse all three sections with data from work carried out by the authors at the Institute of Psychiatry in London from 1986 and 1992. For other reviews, see Allen and Burns (1995), Burns and Forstl (1996), Eisiri (1996) and Howard (1996). PMID- 9332902 TI - What do we really know about late-onset schizophrenia? AB - Actual knowledge on classical late-onset schizophrenia, i.e. the schizophrenic disorders with onset after age 40 years, is reviewed regarding incidence, symptomatology and course. As is shown, sound empirical knowledge is scarce. Reasons for this are, on the one hand, the conceptual and terminological confusion which has occurred internationally regarding this illness group, and, on the other hand, the methodological limitations of the empirical studies conducted on this clinical picture thus far. If we only draw on classical late onset schizophrenia, as originally defined by Bleuler, and primarily on methodologically sound studies, as well as on own studies, we can nevertheless conclude that the term "late-onset schizophrenia" could be omitted. Late-onset schizophrenia does not seem to be a distinct entity, but instead seems to belong to the same illness group as classical schizophrenia with earlier onset. Slight differences in symptomatology and course are probably due to unspecific influences of age. The markedly higher proportion of women among late-onset cases, as well as our finding that symptomatology and course of late-onset women are comparably poor, could possibly be explained by an effect of the female sex hormone oestradiol. PMID- 9332903 TI - Treatment of schizophrenia and delusional disorder in the elderly. AB - With increasing longevity, greater numbers of patients with schizophrenia and delusional disorder will be surviving into advanced age. Antipsychotics form the core of the treatment for both of these psychotic disorders. Treatment of elderly patients with antipsychotics is, however, complicated by a much higher risk of adverse effects such as tardive dyskinesia. More is known about treating patients with schizophrenia than those with delusional disorder. The introduction of newer atypical antipsychotics may herald a new era in the pharmacotherapy of elderly psychotic patients. Nonetheless, judicious dosing is essential in the geriatric population. We discuss the benefits and limitations of the main forms of treatment. PMID- 9332904 TI - Saccadic eye movements and regional cerebral blood flow in schizophrenic patients. AB - This study examined saccadic eye movements, using simple stationary targets, in schizophrenic patients. The targets were eight black points or eight arabic numbered points placed in randomized order on the circumference of a circle. Self paced eye movements during clockwise tracking of these points, by 23 patients and 23 controls, were recorded using an infrared eye-mark recorder. Then the relationship between the saccades and clinical symptoms was investigated. Finally, the relationship between the performance of the saccades and resting regional cerebral blood flow (rCBF) was examined using single photon emission computed tomography with 99mTc-hexamethyl propyleneamine oxime (HMPAO). The results indicate that patients track with significantly fewer correct scores and more deviant scores than controls, in agreement with our previous study. There were two groups of patients: an ordinary group who obtained a full-target-hitting score at a 200-ms setting and a fast group who obtained the full score at 100 ms but not at 200 ms. Some patients displayed significantly more hypermetria than controls. Significant correlations were found between hallucination and delusion symptoms and correct score. With respect to relative rCBF, fast-group patients showed significantly decreased rCBF in the left limbic and inferior parietal areas as compared with ordinary group patients. These findings suggest that some schizophrenic patients view the stationary targets too fast and this may be related to dysfunction in the limbic-parietal association area in the left hemisphere. PMID- 9332905 TI - Interleukin-6-(IL-6) plasma levels in depression and schizophrenia: comparison between the acute state and after remission. AB - The concentration of cytokines such as Interleukin-6 (IL-6) has been reported to be elevated in depressed and schizophrenic patients and, in healthy persons, upon stress. Interleukin-6 plasma levels were determined in depressed (n = 12) and schizophrenic (n = 32) patients during the acute state of illness and after remission at approximately 8 weeks after admission and were compared with healthy controls (n = 12). Patients were diagnosed according to DSM-III-R by the Structured Clinical Interview (SCID). Severity of illness was assessed for depression by the Montgomery Asberg Depression Rating Scale (MADRS) and for schizophrenia by the Brief Psychiatric Rating Scale (BPRS). Interleukin-6 plasma concentrations were elevated during the acute state either of depression or of schizophrenia if compared to controls. After remission, IL-6 concentrations in depressed and in schizophrenic patients had decreased and did not differ significantly from controls. We hypothesize that the elevated IL-6 levels during the acute state of depression or schizophrenia may reflect an unspecific stress response. PMID- 9332906 TI - Use of the selective serotonin reuptake inhibitor citalopram in treatment of trichotillomania. AB - Previous trials of selective serotonin reuptake inhibitors (SSRIs) in the treatment of trichotillomania have provided conflicting data. Furthermore, the efficacy of citalopram, the most selective of the SSRIs, in trichotillomania has not previously been documented. Citalopram was used on an open-label naturalistic basis in 14 (1 male and 13 females) patients who presented with chronic hair pulling and met DSM-IV criteria for trichotillomania. Ratings were completed every 2 weeks for 12 weeks, during which time dosage was increased to a maximum of 60 mg daily (mean dose 36.2 +/- 13.9 mg). One patient was unable to tolerate citalopram. In completers, ratings on each of the scales employed were significantly improved after treatment. Of completers 38.5% were responders (Clinical Global Impressions score of 2 or less) at week 12. Citalopram appears to be safe in trichotillomania, and it may be effective in a subset of patients. Given the relatively low response rate, however, a controlled trial is needed before this agent can be said to be more effective than placebo. The pharmacotherapy of trichotillomania deserves further study. PMID- 9332907 TI - Long-term follow-up of open commissurotomy versus bileaflet valve replacement for rheumatic mitral stenosis. AB - OBJECTIVE: Despite the achievements of third generation mechanical cardiac valve prostheses, conservative procedures are still considered the best surgical option for rheumatic mitral valve stenosis. To compare long-term results of open mitral commissurotomy (Group A) and mitral valve replacement with bileaflet prostheses (Group B) a 15-year follow-up study was carried out. METHODS: From January 1981 to May 1996, 540 consecutive patients with pure isolated rheumatic mitral stenosis underwent mitral valve surgery: 300 had mitral commissurotomy and 240 valve replacement. The follow-up was 99.05% complete and ranged between 1 and 185 months in Group A and from 1 to 171 months in Group B. RESULTS: Hospital mortality was 2% in Group A and 2.08% in Group B. Late mortality was 1% in Group A and 3% in Group B. The 10-year survival rates were 98.7% +/- 1% in Group A and 93.7% +/- 3% in Group B. There was a statistically significant difference of freedom from reoperation in Group B (97.7% +/- 1%) versus Group A (88.1% +/- 2%) (P = 0.04). In group A 14 embolic events occurred (93.7% +/- 2%), and 15 (6.52%) in Group B (83.9% +/- 7%). Haemorrhagic events were observed in 2 patients (0.68%) of Group A (99.3% +/- 0.5%) and in 3 patients (1.3%) of Group B (98.4% +/ 1%). CONCLUSIONS: Long term results of mitral commissurotomy were more satisfactory than those obtained with bileaflet valves. Reoperation rate was higher in mitral commissurotomy. PMID- 9332908 TI - Sinus node function after mitral valve surgery via the transseptal superior approach. AB - OBJECTIVE: The transseptal superior approach can offer an excellent view of the mitral valve but the incision almost always transects the sinus node artery. The purpose of this study was to evaluate the sinus node function after mitral operation by this approach. PATIENTS AND METHODS: We reviewed the electrocardiograms of 76 patients who underwent mitral valve operations either via transseptal superior approach or via right lateral atriotomy. Nine patients who maintained the sinus rhythm for more than one year after surgery via the transseptal superior approach were selected for electrophysiological study to evaluate the sinus node function. RESULTS AND CONCLUSIONS: Postoperative electrocardiographic and electrophysiological studies revealed that the sinus node function after the transseptal superior approach was relatively well maintained for more than one year after the operation. The influence of the transseptal superior approach on the sinus node function in the mid-term postoperative period was apparently mild and did not cause a serious problem. However, some of the patients did show abnormal data in terms of sino-atrial conduction time and intrinsic heart rate. Therefore, further follow-up of the sinus node function is necessary in patients who underwent mitral surgery through the transseptal superior approach. PMID- 9332909 TI - Surgery of chest wall deformities. AB - OBJECTIVE: To evaluate the medium-term results of 77 surgical corrections in patients with chest wall deformities, 53 (68.8%) with pectus excavatum and 24 with pectus carinatum, operated upon from 1985 to 1994. METHODS: The mean age of the patients was 14.7 years (4-39 years) and 77% were younger than 15 years of age. There were 59 male (76.7%) and 18 female patients. Only four had a family history of the malformation. Seven patients (9.1%) presented with asthma-like symptoms, and 13 (16.9%) referred dyspnea and tiredness for small efforts. The remainder (74.2%) were asymptomatic, but most were psychologically disturbed by the deformity and postural abnormality. Two patients had other skeletal abnormalities. The modified surgical technique used in all cases consisted of subperichondrial resection of the abnormal costal cartilages, transverse and longitudinal osteotomies of the sternum and internal stabilization with a steel rod which was generally removed between 6 and 12 months postoperatively. RESULTS: There was neither early nor late mortality. One patient had a pneumothorax which required chest tube drainage. The mean admission time was 10.5 days (8-14 days). Follow-up was complete, and 90% of the patients had increased effort tolerance. Five of the seven patients (72%) with 'asthmatic' symptoms showed a decrease in the frequency of the crises. Two patients had recurrence of the depression by 3 and 8 months, respectively. The remaining 75 patients (97.3%) were satisfied with the cosmetic result of the surgery. CONCLUSIONS: Surgical treatment of chest wall deformities using this technique leads to good cosmetic, orthopedic and psychological results. We believe that the operations should be performed at any age in patients who have at least a moderate deformity. PMID- 9332910 TI - Life-saving muscle flaps in tracheobronchial dehiscence following resection or trauma. AB - OBJECTIVE: In the presence of acute inflammation and necrosis of the wall, tracheo-bronchial defects are difficult to manage. The absence of adequate vascularization and the contaminated area prevent successful direct re-suturing. METHODS: In order to restore a sufficient blood supply we used a pedicled latissimus dorsi or a pectoralis major flap that was entered into the thorax after a 10-cm resection of the second rib. A portion of the muscle was fitted into the tracheo/bronchial defect by reinforced sutures. The remaining muscle was sutured to the tissue surrounding the defect. This method was applied in various septic conditions: Bronchial defects; complete dehiscence of the right (n = 6) or left (n = 1) main bronchus at the carinal level following resection for lung cancer (n = 4) or for tuberculous (n = 2) on nontuberculous pleuropneumonia (n = 1). Tracheal defects; (1) destruction of one third of the tracheal circumference involving the cricoid down to the fourth ring following tracheotomy in presence of a septic sternum after intrathoracic goiter and Bechterew's disease; (2) 30% dehiscence of the anastomosis and septic sternum following tracheal resection; (3) Mediastinitis involving tracheal and esophageal wall following a 7 cm long iatrogenous laceration of the intrathoracic trachea. RESULTS: In one case the latissimus dorsi developed venous stasis on day 2 and was replaced by the pectoralis major muscle which showed uneventful healing. In all other patients the muscle flap resulted in an uneventful closure of the defect and recovery. CONCLUSIONS: Large, well vascularized, pedicled muscle flaps ensure a safe closure of tracheo-bronchial defects or dehiscences even in presence of gross necrosis and sepsis. PMID- 9332911 TI - Diagnostic and therapeutic video assisted thoracic surgery (VATS) following chest trauma. AB - OBJECTIVE: Thoracic injury remains a major source of morbidity and mortality in urban trauma centers. With the advent and increasing expertise in video assisted thoracic surgery, this modality has become an attractive alternative in the management of patients with thoracic injury. This report will review our experience with video assisted thoracic surgery at a level I trauma center and attempt to further delineate the indications for and timing of thoracoscopy in thoracic trauma. METHODS: We identified 16 patients who had undergone video assisted thoracic surgery following chest trauma between July 1991 and June 1994. There were 15 penetrating and one blunt trauma. All 16 patients were initially treated with tube thoracostomy. From 0-20 days post-injury, video assisted thoracic surgery was attempted with either diagnostic or therapeutic intentions. RESULTS: Twelve of the 16 patients (75%) had successful thoracoscopy. Three patients had diaphragmatic injury excluded and nine patients had successful evacuation of clotted hemothoraces. Evacuation of clotted hemothorax up to 7 days post-injury was safe and easily accomplished. Four patients (25%) had unsuccessful thoracoscopy and were converted to standard thoracotomy; failure was attributed to either suboptimal single lung ventilation or severe pleural inflammatory reaction. The only death in the entire group occurred 10 days after a thoracotomy for retained hemothorax. The median post-operative hospital stay following successful video assisted thoracic surgery was 3.5 days. CONCLUSIONS: Video assisted thoracic surgery can be utilized as an effective and safe method for the initial diagnostic evaluation and surgical management of stable patients with penetrating thoracic trauma. PMID- 9332912 TI - Proposal for improved staging criteria for carcinoma of the esophagus and cardia. AB - OBJECTIVE: Current staging for carcinoma of the esophagus and cardia remains imprecise. In an effort to improve on presently accepted staging criteria, new and improved criteria were sought. METHODS: A total of 408 specimens resected for carcinoma of the esophagus or cardia between January 1, 1970, and January 1, 1994, were available for analysis. Pathology reports were reviewed, and available histologic slides were examined microscopically. When necessary, paraffin blocks of excised specimens were recut for further pathologic evaluation. On the basis of these findings, tumors were staged according to the criteria of American Joint Committee on Cancer (AJCC). New criteria were established based on the WNM concept and staged accordingly. Survival rates based on these sets of criteria were calculated for each stage, and results were compared. RESULTS: Because our previous studies had shown no advantage provided by the revised AJCC criteria compared with those originally proposed, we modified the WNM system by eliminating the subdivisions of Stage II, reducing the T categories by 1, T3 and T4 having shown no survival differences, and increasing the N categories by 1, depending on the number of nodes involved, e.g. NO = no positive nodes; N1 = 1-4 positive nodes, and N2-5 or more positive nodes. The resulting staging system and 5-year survival rates obtained thereby are as follows: Stage O (TO, is, 1 NO), 88.2%; Stage I (T1N1, T2NO), 50.3%; Stage II (T2N1, T3N0) 22.5%; Stage III (T3N1, any T N2), 10.7%; and Stage IV (M1) 0%. CONCLUSIONS: A new staging scheme for carcinoma of the esophagus and cardia is proposed that provides better prognostic stratification of patients than existing ones. PMID- 9332913 TI - Concurrent radio-chemotherapy in N2 non small cell lung cancer: interim analysis. AB - OBJECTIVE: In recent years many authors have been focused on N2 non-small cell lung cancer patients to determine whether the rate of resectability and long term survival can be improved by a combined preoperative treatment, with significant results. Following these experiences, we planned an induction therapy trial to assess the impact on downstaging, resectability and survival of concurrent radio chemotherapy on N2 non-small cell lung cancer patients. METHODS: Between January 1990 and August 1995, 82 N2 non-small cell lung cancer patients (44 IIIA and 38 IIIB) received preoperative chemo-radiotherapy with a single cycle of Carboplatin (90 mg/m2 per day for days 1-4), concurrent with radiotherapy (daily radiation dose of 180 cGray for a total of 5040). After surgery, all patients received multi-drug chemotherapy with Carboplatin 300 mg/m2 per day on day 1 and VP-16 100 mg/m2 per day on days 1, 2 and 3, for a total of 6 monthly cycles. Patients with unresectable tumors underwent to this multi-drug chemotherapy, directly. RESULTS: Two patients were excluded from the study. When the remaining 80 patients had a 'clinical' re-staging, 41 (51.3%) showed a major response, 36 (45%) had minimal or none response, and 3 (3.7%) had progression of disease. Forty-one patients were judged to be resectable, 11 staged IIIB, and 30 IIIA; 2 patients of the IIIB group refused surgery. Of the 39 operated cases, 37 were completely resected (resectability rate: 94.8%). We report one perioperative death due to respiratory failure and two major complications. The overall actuarial 5 year survival is 24.5%. Downstaging was observed in 22 patients (56.4%), with three patients (7.7%) having no evidence of tumor in the specimen, 16 (41%) having sterilization of all lymph nodes, and three (7.7%) having sterilization of mediastinal nodes but positive N1 nodes. The 5-year actuarial survival is 53% for patients who had complete resection and 0% for patients with no resection (P = 0.0000). CONCLUSIONS: The following conclusions are possible: preoperative radiotherapy and chemotherapy with Carboplatin is well tolerated by patients, does not increase postoperative complications and produces an high rate of response. There is an high resection rate for patients who respond to the therapy. Patients with major response who undergo complete surgical resection had statistically significant improved survival compared with patients whose disease was not resected. PMID- 9332914 TI - Time trends and survival after surgery for p-stage IIIa, pN2 non-small cell lung cancer (NSCLC). AB - OBJECTIVE: To evaluate the role of surgery for p-stage IIIa, pN2 non-small cell lung cancer (NSCLC), time trends and survival after surgery and the prognostic factors were investigated retrospectively. METHODS: Consecutive patients, 155, with p-stage IIIa, pN2 NSCLC who underwent thoracotomy at the Department of Thoracic Surgery, Chest Disease Research Institute, Kyoto University between January 1976 and December 1990 were divided into three groups by the period of operation (the earlier period: 1976-1980, n = 49; the middle period: 1981-1985, n = 55; and the later period: 1986-1990, n = 51), and were reviewed. Of the 155 patients, 84 (54.2%) were preoperatively evaluated to have mediastinal lymph nodes metastases (cN2 disease). RESULTS: The 5 year survival rates in the earlier, middle and later periods were 12.1, 18.6, and 43.8%, respectively, showing significant improvement in the later period (P < 0.001, for the later period versus the earlier period or the middle period). The improvement was caused by decrease in the rate of operation-related death (4.1, 1.8, and 0.0%, in the earlier, the middle, and the later period, respectively), increase in the rate of complete tumor resection (59.1, 76.4, and 96.1%, respectively), and decrease in the ratio of pT3N2M0 patients (44.9, 34.5, and 17.6%, respectively) having poor prognosis compared with pT1-2N2M0 patients. Decrease in the ratio of cT3N2M0 patients and for increase in the rate of complete resection could be realized by accurate preoperative diagnosis with introduction of chest computed tomography (CT). Based on the preoperative evaluation, the 5 year survival rates of cT1N2M0, cT2N2M0, and cT3N2M0 patients were 39.4, 30.5, and 10.2%, respectively, showing significant poor prognosis in cT3N2M0 patients. CONCLUSION: In cT1-2N2M0 or pT1-2N2M0 patients, a good prognosis can be realized by complete tumor resection with mediastinal lymph nodes dissection. In contrast, surgical treatment should not be justified in cT3N2M0 or pT3N2M0 patients. PMID- 9332915 TI - Disease recurrence after resection for stage I lung cancer. AB - OBJECTIVE: To asses the incidence of local recurrence and distant metastases after complete resection for stage I lung cancer in order to predict the predominant prognostic factors. METHODS: We retrospectively reviewed 123 patients with stage I lung cancer who underwent curative resection over a 2-year period between January 1987 and December 1988. There were 83 male and 40 female patients with a mean age of 64.8 +/- 12 years (range between 39 and 82 years). Multivariate analysis of prognostic factors for long term survival was undertaken. RESULTS: T1N0 lesions were found in 34 patients and T2N0 in 89. The histological diagnosis was Squamous carcinoma in 75, Adenocarcinoma in 38, large cell carcinoma in 6 and small cell carcinoma in 4 patients. Pneumonectomy was performed in 27 patients (5 T1 and 22 T2) while 96 required lobectomy (29 T1 and 67 T2). At 5 years 50 patients died. This was due to local recurrence in 12, distant metastasis in 24, second primary in 1, unrelated disease in 3, while the cause was unknown in 10 patients. At 5 years, 10 patients were alive with evidence of recurrence. The mean interval for local recurrence was 19.8 months and for distant metastasis was 18 months. The overall 5 year survival was 67% +/- 4 for T1 and 56% +/- 5 for T2 lesions (NS). The rate of recurrence was significantly less for T1 lesions (P = 0.02). Survival was significantly less for patients requiring pneumonectomy rather than lobectomy (P = 0.01) whether for T1 or T2. CONCLUSION: In stage I lung cancer T2 lesions requiring pneumonectomy for complete resection had a worse prognosis and higher incidence of local recurrence. PMID- 9332916 TI - Should the number of pulmonary metastases influence the surgical decision? AB - OBJECTIVE: To assess, using a large homogeneous retrospective series, the prognostic value of the number of resected pulmonary metastases, and thus, to determine to what extent the number of resectable metastases should influence the surgical decision. METHODS: The survival analysis of all patients operated on for pulmonary metastases at a single center, the comparisons of 2 'histologic' groups (sarcoma and carcinoma) and, within each histologic group, of three subgroups with different numbers of resected metastases (1, 2-4, and > or = 5) were performed. The log-rank test was used to compare survival curves. RESULTS: Among 575 adult patients operated on with curative intent before December 1991, the first operation allowed the complete resection of a known number of histologically proven viable pulmonary metastases in 230 and 151 patients with metastases from carcinoma and sarcoma, respectively. The 5- and 10-year probabilities of survival (Kaplan-Meier) were 37 and 23%, respectively in carcinoma patients, and 31 and 28%, respectively in sarcoma patients (log-rank test: ns). Only the difference between patients with 1 versus 2-4 metastases from carcinoma proved statistically significant (P = 0.02), with 5-year survival estimates of 41 and 25%, respectively. Beside survival, the only significant difference between the subgroups of patients with different numbers of resected metastases was the mean interval between the diagnosis of pulmonary metastases and the resection of pulmonary metastases, which was significantly longer in patients with several metastases in both histologic groups. CONCLUSIONS: In patients with resectable pulmonary metastases from sarcoma or carcinoma, the number of metastases should have little influence on the surgical decision, except for delaying this decision in patients with several metastases until a significant interval, with or without treatment, has shown that metastatic disease remains resectable and confined to the lungs. PMID- 9332917 TI - Left internal mammary elongation with inferior epigastric artery in minimally invasive coronary surgery. AB - OBJECTIVE: Sometimes the left internal mammary artery (LIMA) is not long enough to reach a too lateral LAD when a left anterior small thoracotomy (LAST operation) is the surgical approach to graft the LAD. LIMA elongation with an inferior epigastric artery (IEA) can be an useful surgical option. METHODS: From November 1994 to June 30, 1996, out of 289 patients who underwent LAST operation; 28 patients had a LIMA elongation with an IEA, 20 patients had single vessel disease, 4 had two vessel disease, and 4 three vessel disease. Mean age was 62 +/ 22 (48-84) and mean EF was 57 +/- 86. The IEA was used only when the LAD was totally or nearly occluded with no transmural myocardial infarction (high expected run off). RESULTS: All patients had an uneventful recovery. After 315 +/ 104 days from surgery all were asymptomatic. A late doppler flow assessment, performed in 28 patients, showed a high velocity diastolic flow in 27. One patient was reoperated on because of graft occlusion 84 days after surgery. An angiography was performed after 87.5 +/- 23.3 days in 22 patients. All conduit and anastomoses were patent but one, (patency rate 21/22, 95.4%); another showed mild anastomotical stenosis at the LIMA-IEA junction without clinical signs (perfect patency rate 20/22, 90.9%). CONCLUSIONS: IEA elongation of LIMA is an alternative strategy to reach a lateral LAD in selected cases; a satisfying patency rate can be expected, when correct surgical indications are used. PMID- 9332918 TI - Lateral MIDCAB grafting via limited posterior thoracotomy. AB - OBJECTIVE: Minimally invasive direct coronary artery bypass (MIDCAB) is a technique for coronary artery bypass grafting performed under direct vision without sternotomy or cardiopulmonary bypass. The approach has been used principally for primary single vessel grafting of the anterior or inferior coronary circulation. This initial experience presents a new lateral technique for patients with isolated circumflex coronary disease which can be used for both primary and reoperative revascularization with either saphenous vein or a free radial artery conduit. METHODS: Lateral MIDCAB grafting of the circumflex coronary circulation was accomplished over a 33 month period at a single center using saphenous vein or free radial artery as the bypass conduit. Through a limited posterior thoracotomy, the lung is deflated and reflected superiorly. The pericardium is opened below the phrenic nerve to expose an obtuse marginal branch of the circumflex coronary artery. After heparinization, the coronary artery is temporarily occluded proximally and distally with local immobilization and an arteriotomy is performed. The distal anastomosis with running suture is followed by the proximal anastomosis on the descending aorta below the hilum of the lung using a side-biting clamp and radiopaque marker. Intraoperative transit time ultrasound flow measurements confirm adequate graft flow before wound closure. RESULTS: To date, 19 patients have undergone this procedure with a mean follow-up of 12 months. A total of 12 patients received saphenous vein grafts and 7 patients received radial artery grafts. There was one death from arrhythmia on postoperative day 9. There was one elective conversion to conventional sternotomy due to inadequate exposure. Graft flows averaged 33.3 cc/min (range 5-87) and the mean postoperative length of stay was 4.5 days; 4 patients underwent recatheterization; 1 had graft occlusion and 2 received late postoperative catheter-based interventions. All patients are currently free of symptoms. CONCLUSIONS: Lateral MIDCAB grafting provides focused revascularization to the circumflex distribution in both primary and reoperative settings. This approach avoids the hazards of resternotomy, eliminates cardiopulmonary bypass, and hastens postoperative recovery. These early results suggest the technique is effective at relieving symptoms and minimizing perioperative morbidity. Further experience at multiple centers will serve to define the ultimate capabilities of this new approach. PMID- 9332919 TI - Less invasive off-pump CABG using a suction device for immobilization: the 'Octopus' method. AB - OBJECTIVE: Target site immobilization is essential to enable meticulous anastomosis suturing during coronary artery bypass grafting on the beating heart. A novel device ('Octopus') was developed for local heart muscle immobilization by suction. The purpose of this study was to investigate the efficacy of the method through a limited access. METHODS: The suction device, placed on either side of the recipient coronary artery and fixed to the operating table-rail through an arm construction, restrains anastomosis site motion to 1 x 1 mm. A total of 27 patients underwent off-pump arterial bypass grafting using this method. Preoperatively, all patients had angina class III (NYHA) and were failed or unsuitable candidates for balloon angioplasty. Surgical access was via a 10-cm anterior thoracotomy (n = 26) or 10-cm subxiphoid incision (n = 1). RESULTS: Harvesting of the graft required 48 +/- 12 min (mean +/- S.D.). Immobilization with the 'Octopus' was effective and facilitated precise anastomosis suturing of 20 single and 7 sequential grafts. Immobilization did not change cardiac index and mean arterial blood pressure. During coronary surgery, however, inotropic drug support was used in 5 of 27 (18%) of patients. There was no myocardial infarction. Only minor transient complications were met. There were electro cardiographical signs of pericarditis in 6 patients. The postoperative hospital stay ranged from 2 to 6 days, mean 4.0 +/- 1.2 days. The mean follow-up is 6.5 +/ 4 months (range, 1-12 months). All patients except one were in functional class I without angina. Social activities were resumed within 4 weeks. At 6 months angiography was performed in 15 out of 27 patients. The patency rate of 19 out of 20 anastomoses was 95%. All distal grafts were patent. One side to side anastomosis was occluded. CONCLUSIONS: The 'Octopus' immobilization method is safe and effective. It facilitates less invasive CABG in selected patients and gives way to fast recovery by reducing invasiveness. PMID- 9332920 TI - Single aortic cross-clamp technique in coronary surgery: a prospective randomized study. AB - OBJECTIVE: To test the hypothesis of an improved myocardial and cerebral protection by combining blood cardioplegia and the single aortic cross-clamp technique, 100 patients were enrolled in a prospectively randomized study and stratified for preoperative conditions. METHODS: In Group I, 55 patients underwent myocardial revascularization using crystalloid cardioplegia and the conventional partial occluding clamp technique to perform proximal anastomoses, whereas in Group II, 45 patients were operated on combining blood cardioplegia and the single aortic cross-clamp technique. Unstable angina, emergency procedures, reoperations and preoperative counterpulsation accounted for an higher risk score in group II patients (P < 0.03). Operations were performed by the same surgical team. Aortic cross-clamp time was significantly longer in group II patients (59 +/- 22 vs. 47 +/- 18 min.) (P < 0.001). Other intraoperative variables were not significant. RESULTS: A 70-year-old male in group I died on post-operative day 5 as a consequence of a major neurological event. Length of ventilatory dependency, post-operative bleeding, need for blood transfusions, ICU stay, and hospital stay were similar between the two groups (P = NS). Patients in group I showed a strict correlation between the duration of surgical ischemia and post-operative myocardial necrosis. Analysis of combined mortality and morbidity events (adverse events) between the two groups, led to a significant prevalence in group I patients (P < 0.03) in spite of an higher pre-operative risk score and longer ischemic times in group II patients. Neurological lesions remained confined to group I patients. CONCLUSIONS: The combined use of blood cardioplegia, delivered via the antegrade and retrograde routes, and the single clamp technique to perform myocardial revascularization, might enhance myocardial and cerebral protection when compared to conventional methods. Larger groups of patients are needed to support this trend. PMID- 9332921 TI - Cardioprotective effects of diltiazem infusion in the perioperative period. AB - OBJECTIVE: To evaluate the perioperative effects of intravenous diltiazem infusion on left ventricular functions, hemodynamics and as an anti-ischemic and antiarrhythmic agent in patients undergoing coronary artery bypass grafting (CABG). METHODS: A double blind, randomised study was performed on 71 patients undergoing elective CABG. Infusion of diltiazem (0.1 mg/kg per h, n = 34) or nitroglycerin (1 microgram/kg per min, n = 37) was given for 24 h starting from onset of cardiopulmonary bypass. Holter monitoring, electrocardiogram and serum cardiac enzymes levels were used to diagnose myocardial ischemia. Myocardial function was assessed by perioperative transesophageal echocardiography. RESULTS: The two groups did not differ with respect to preoperative and operative data. Diltiazem had no influence on hemodynamic parameters except for significant reduction in post operative heart rate and pulse pressure rate. Transient ischemic events (dilitiazem 10.2% versus nitroglycerin 33.3%, P = 0.15) and transient coronary spasm (diltiazem-6.8% versus nitroglycerin 25.9%, P = 0.15) were reduced in the diltiazem group as compared with the nitroglycerin group. The postoperative incidence of atrial fibrillation (diltiazem 3% versus nitroglycerin 22%, P = 0.03), supra ventricular tachycardia (diltiazem-3% versus nitroglycerin 22%, P = 0.03) and average ventricular premature contraction per h (diltiazem 40.2 +/- 10.2 versus nitroglycerin 53.8 +/- 12.3, P < 0.01) were significantly lower in the diltiazem group. Transesophageal echocardiography showed no significant difference in left ventricular functions and better preservation of left ventricular diastolic functions in post cardiopulmonary bypass period in diltiazem group. In addition mean deceleration time for the E wave on a 12 h post cardiopulmonary bypass period was significantly lower in the diltiazem group as compared with nitroglycerin (diltiazem 131 +/- 6 versus nitroglycerin 171 +/- 6, P < 0.01). CONCLUSION: The present study demonstrates that diltiazem infusion provides superior anti-ischemic protection and control of supraventricular arrhythmias as compared to nitroglycerin and does not produce any negative inotropic effect, as demonstrated by transesophageal echocardiography. PMID- 9332922 TI - Assessment of human long saphenous vein function with minimally invasive harvesting with the Mayo stripper. AB - BACKGROUND: The use of the Mayo Stripper to harvest the long saphenous vein has been shown to improve morbidity from leg wound incisions. It has not been universally accepted because of a perceived increase in injury to the venous conduit. OBJECTIVE: To compare the function of undistended autologous long saphenous vein harvested by a Mayo stripper with the traditional 'open' technique in the same patient (n = 12) appearance. METHODS: Vascular reactivity was assessed in isolated organ baths. Contractile function was measured in response to increasing concentrations (10(-9)-10(-5) mol) of 5-hydroxytryptamine and noradrenaline. This was calculated as a percentage of the maximum contractile response to 90 mM KCl measured in millinewtons (mN) (control 41.4 +/- 12.1, (n = 11), open technique 35.8 +/- 11.1, (n = 11), Mayo stripper 33.7 +/- 15.9, (n = 11)). The endothelial dependent and independent function was assessed with acetylcholine and sodium nitroprusside, respectively. RESULTS: There was no significant difference in response to both constrictors and dilators between vein taken with the Mayo stripper compared with the traditional open technique (n = 6 for each observation; P > 0.05 by ANOVA). Histological examination by light microscopy of the vessel segments removed with the Mayo stripper was unable to show any significant damage to the vessel wall. Both functional and morphological studies were conducted by 'blinded' observers. One-year follow-up with magnetic resonance angiography (MRA) and stress thallium tomography demonstrated a patency rate with lower and upper estimates of 80 and 94%. CONCLUSIONS: We have shown that harvesting the long saphenous vein with a Mayo stripper does not compromise vascular reactivity of the long saphenous vein or long-term patency. PMID- 9332923 TI - What in-vitro method should surgeons use to evaluate the clinical behavior of arterial bypass conduits. AB - Surgeons have traditionally relied on ring preparations to predict how arterial bypass conduits will behave in the postoperative circulation. OBJECTIVE: This study compared pharmacologic [norepinephrine (NE) challenge] and physiologic [arterial preload] responses of gastroepiploic (GEA) and internal thoracic (ITA) arteries in a standard static ring preparation and a dynamic perfusion system. METHODS: Six GEAs (1.0-1.5 mm dia.) and six ITAs (1.5-2.0 mm dia.) 11 cm long were harvested from adult pigs and mounted on a computer controlled perfusion system. Inflow pressure was set at 80 mmHg and outflow resistance was adjusted to simulate high (80-90 ml/min) and low (15-20 ml/min) flow demands. NE response (10(-9)-10(-5) M) was measured under low flow conditions and at high flow conditions when distal arterial pressure (load) was reduced. NE response (10(-9) 10(-5) M) was also evaluated in arterial rings (ITA N = 6, GEA N = 6) with tensions adjusted to simulate the loads occurring at low flow (80 mmHg) and high flow (60 mmHg) situations. RESULTS: In the static ring preparation, NE response [ED50] was similar for both GEA and ITA and was not affected by load. The dynamic preparation demonstrated that the GEAs were significantly more responsive to NE than the ITAs [ED50 high flow ITA 6.1 +/- 0.3**, GEA 7.2 +/- 0.3***; *P < 0.05 versus baseline, **P < 0.05 versus low flow values, ***P < 0.05 versus ITA]. Furthermore, in the dynamic preparation, NE response was profoundly affected by reduced load which occurs under high flow conditions [7.18 +/- 0.3 versus 6.1 +/- 0.3 under high flow and 5.8 +/- 0.1 versus no response under low flow conditions]. CONCLUSION: Static ring preparations do not discern differences between ITA and GEA susceptibility to spasm and fail to detect the effect of load. The dynamic preparation demonstrated significant differences between the GEA and ITA potential to spasm which is consistent with widespread clinical experience. Furthermore a dynamic preparation is highly sensitive to reduced load which occurs under high flow conditions. Although it is more demanding, the dynamic preparation provides superior information to the surgeon in predicting the behavior of arterial bypass grafts. PMID- 9332925 TI - Surgical closure of muscular ventricular septal defects using double umbrella devices (intraoperative VSD device closure). AB - OBJECTIVES: Surgical closure of some muscular ventricular septal defects has been proven to be difficult. In order to simplify the surgical technique we have used intraoperatively Rashkind double umbrella devices to occlude muscular ventricular septal defects. METHODS: On the basis of haemodynamic and echocardiographic study five children aged 4, 6, 7, 14 and 41 months were considered suitable candidates for intraoperative closure of muscular ventricular septal defects (midmuscular in three cases, apical in two) by Rashkind devices. Three of them had previously undergone pulmonary artery banding at 10, 11 and 41 days, respectively. During hypothermic cardiopulmonary by pass a delivery system was introduced across the tricuspid valve into the right ventricle and then passed through the ventricular septal defect; the distal umbrella of a 17 mm device was opened in the left ventricular cavity; a traction was applied to the introducer and the proximal umbrella was opened on the right side straddling the interventricular septum; the device was then secured on the right side by few stitches. In one case because of the wide diameter of the ventricular septal defect two umbrellas were used. The surgical procedure was completed with debanding and/or closure of other defects close to the aortic or tricuspid valve. RESULTS: Immediate results, tested by epicardial or transesofageal echo, showed a minimal residual shunt in 4 patients and a moderate shunt in one. No early deaths occurred. A complete atrioventricular block developed in 1 patient who had an additional perimembranous defect closed with a prosthetic patch: a permanent pace maker was inserted 3 months after the operation. There was a late death for untractable right ventricular failure in 1 patient who had a large residual shunt erroneously considered moderate. In this patient, the size of the defect was underestimated both preoperatively then intraoperatively. The four survivors are doing well with no signs of hemodynamically significant residual shunts. CONCLUSIONS: The use of Rashkind umbrella devices for closing intraoperatively muscular defects can be helpful to standard surgical techniques when technical problems make patch closure difficult. Its use avoid the need of left ventriculotomy. Careful definition of the size of the defect is mandatory to select suitable candidates. PMID- 9332924 TI - Mediastinitis after aorto-coronary bypass surgery. AB - OBJECTIVES: To identify risk factors in 60 cases of mediastinitis amongst 2512 patients (2.3%) subjected to isolated coronary bypass surgery from March 1988 through December 1995, treated by a closed irrigation/drainage system. PATIENTS AND METHODS: The mean age of the 60 patients was 56.9 +/- 6.8 years (45-81 years) and 55 (91.6%) were male. Early mediastinal reexploration was performed in all cases immediately after the diagnosis of mediastinitis, with debridement of necrosed tissues, followed by implantation of a closed-circuit irrigation system of the mediastinum constituted by irrigation catheter and drain, closure of the sternum and skin, and specific systemic antibiotic therapy. The mean interval between the original surgery and reexploration was 9.4 days (range 6-14 days). No patient required more extensive procedures, namely omental or muscular flaps. Twenty potential risk factors in patients with mediastinitis, including diabetes mellitus, obesity, coexistence of peripheral vascular disease, decreased LV function, use of inotropes, mediastinal blood drainage and utilization of double IMA, were compared with the group without mediastinitis. RESULTS: Mean cardiopulmonary bypass time was 74.1 +/- 8.1 min, anesthetic time 3.5 +/- 0.8 h and postoperative mechanical ventilation 18 +/- 3 h. A total of 23 patients (38.3%) received one IMA and 35 (58.3%) two IMAs. In the postoperative period, 7 of the 60 patients (11.6%) had required inotropes because of low output. Mediastinal blood loss was 1112cc +/- 452cc and 9 patients (15%) were transfused. Cultures were positive in 40 cases (66.6%) and the most frequent infecting agent was the Staph. epidermidis in 25 cases (62.5%), followed by Candida albicans and Enterobacter and Serratia species (7.5% each); 1 patient (1.7%) died and 9 (15%) had renal failure. The irrigation/drainage was maintained for a mean of 9.1 days (5-83 days). Patients with mediastinitis had a significantly higher prevalence of diabetes (41.6% vs. 18.8%; P < 0.01), obesity (48.3% vs. 15.2%; P < 0.001), peripheral vascular disease (11.6% vs. 4.0%; P < 0.05), but a lower incidence of poor LV function (18.3% vs. 32.7%; P < 0.05). A double IMA was used more frequently in patients who had mediastinitis (58.3% vs. 23.5%; P < 0.001) CONCLUSIONS: Diabetes mellitus, obesity, co-existence of peripheral vascular disease and use of double IMA are risk factors for mediastinitis after coronary artery surgery. The efficacy of the closed method of treatment with a mediastinal irrigation/drainage system was increased with early diagnosis and reintervention. PMID- 9332926 TI - Pulmonary artery augmentation with autologous aortic tissue. AB - OBJECTIVE: To assess durability and viability of autologous aortic tissue used to augment severe branch pulmonary artery stenosis with a novice surgical technique. PATIENTS AND METHODS: Seven patients underwent corrective surgery for complex cyanotic congenital heart disease. Their age ranged from 3-6 years, and their weight 11-17.4 kg. All had concomitant branch pulmonary artery stenosis repaired utilizing an autologous patch, harvested from the patient's own aorta by excising a ring and opening it to form the patch. The aorta is reconstructed directly by end to end anastomosis. RESULTS: One patient died in hospital. Another patient died at 18 months at home. The surviving five patients have remained well in the follow up period of mean 31 months (range 10-52). All patients were restudied by follow up echocardiography and remain with no evidence of the aortic autograft tissue calcification or stenosis. The reconstructed aorta showed no stenosis at the site of anastomosis. CONCLUSION: The intermediate term results of this novice surgical technique appear encouraging and justify the technique. However, longer follow up will be required to confirm the continued growth of this patch material. PMID- 9332927 TI - Results of left atrioventricular valve reconstruction after previous correction of atrioventricular septal defects. AB - OBJECTIVE: The objective of this study was to determine causes of severe left atrioventricular (AV) incompetence and the factors leading to the success of valve repair later after correction of atrioventricular septal defects (AVSD). METHODS: A total of 28 patients aged 5 months to 38 years (mean age 6.7 years) were operated for significant (grade II-III) left AV valve incompetence (LAVVI), two months to twenty-five years (median 1.5 years) after correction of complete (11 patients) or partial atrioventricular septal defects. Fourteen patients had initially undergone surgery during infancy. RESULTS: At reoperation a completely open or partially sutured cleft was found in 16 patients combined with dysplastic valve tissue in four cases, with a fibrotic valve in three cases, with posterior leaflet prolapse in two cases, with a double orifice valve in three cases, and a parachute valve in two cases. Partial or complete reopening of a previously sutured cleft caused by suture dehiscence was found in 12 cases combined with a fibrotic valve in five cases, with a dysplastic valve in one case and with severe deformity of valve in one case. A combination of these anomalies was observed in seven patients in both groups. Left atrioventricular valve repair including cleft closure combined with annuloplasty and other surgical procedures resulted in the disappearance or significant diminishing of LAVI in 18 patients (64%). Severe SAVI persisted in six patients, five of them exhibiting a combination of several additional left AV valve anomalies (fibrotic or dysplastic valve, parachute valve). Five of these six patients underwent successful left AV valve replacement with a mechanical prosthesis 7 days to 2 years after reoperation. The presence of additional left AV valve anomalies was the single statistically significant factor for recurrent major LAVVI after reoperation (P = 0.0106). There were two postoperative deaths in patients with mild LAVVI after surgery, and no late deaths. CONCLUSION: An open cleft is the major factor of late severe SAVVI after correction of AVSD. Although suturing the cleft in conjunction with performing annuloplasty improved valvular function in most of the cases, the presence of severe left AV valve anomalies increased the risk of recurrent LAVVI and the need for valve replacement, thus playing a major role in determining the outcome of valve reconstruction in patients after reoperation. PMID- 9332928 TI - The interrupted aortic arch: an overview after 20 years of surgical treatment. AB - OBJECTIVE AND METHODS: The records of 95 patients with interrupted aortic arch, admitted to our center from 1975 to 1995, were reviewed. We were particularly interested in the long term results and evaluated the impact of the preoperative state on the outcome after surgery. RESULTS: Using the 'Celoria and Patton' classification, 13% were type A, 84% type B and 3% type C. Among various associated anomalies were ventricular septal defects and left ventricular outflow tract obstructions, either subvalvular or due to a hypoplastic annulus or a bicuspid valve. We have also seen complex malformations such as truncus arteriosus communis, double outlet right ventricle and transposition of the great arteries. Preoperative neurological disorders, among them the Di George's syndrome, were found in 29% of the cases. Our long term results show 52 patients to be alive, of which 89% are in good clinical condition. Due to improved operative techniques and changes in the management of neonates respectively, early mortality was 17% between 1985 and 1995 compared to 42% between 1975 and 1985. Reoperations were necessary due to arch stenosis, compression of the bronchus or left ventricular outflow tract obstruction. CONCLUSIONS: Nevertheless, mortality after surgical repair of an interrupted aortic arch has dropped significantly and the preoperative condition plays an important role in the outcome. Sepsis, low output, low weight (under 2400 g), severe left ventricular outflow tract obstruction and complex malformations impeded surgery in 13% of cases. Immediate surgical intervention is the only therapy. Arch continuity and repair of associated anomalies could be achieved in the remaining collective. Most of the children have a good quality of life. The preoperative condition seems to influence late neurological disorders. PMID- 9332929 TI - The time course of pulmonary transfer factor changes following heart transplantation. AB - OBJECTIVE: The pulmonary transfer factor for carbon monoxide (TLCO) has been reported to decline following heart transplantation, but the time course of this decline is not well documented. The aim of this study was to define the longitudinal changes in TLCO after heart transplantation. METHODS: Single breath TLCO, lung volumes and expiratory flow rates were prospectively measured in 57 patients (mean age 49 years, range 19-61) before and at least once after heart transplantation. Thirty seven of the 57 patients had four post-transplant assessment which were performed at 6 weeks, 3, 6 and 12 months in 26 patients and at 12, 18, 24 and 36 months in 11 patients. Results were compared with data from 28 normal subjects (mean age 40 years, range 19-61). RESULTS: Before transplantation there was a mild impairment of lung volumes and expiratory flow rates. At 6 weeks after transplantation, there was a further reduction in the forced expiratory volume in one second, forced vital capacity, residual volume and total lung capacity, but all of these increased in the subsequent measurements to exceed their pre-transplant values at about 1 year after transplantation. Haemoglobin-corrected TLCO was also reduced before transplantation compared to normal controls (74.3% and 98.6% of predicted respectively, P < 0.001). Although TLCO per unit alveolar volume (KCO) was relatively preserved in heart transplant candidates, it was still significantly lower than that of normal controls (92.6% and 105.3% of predicted respectively, P < 0.05). After transplantation, mean haemoglobin-corrected TLCO and KCO declined by 12% and 20% of predicted respectively) with the majority of patients having reductions greater than 10% of predicted. The decline in TLCO and KCO was evident at 6 weeks after transplantation with no further changes in the subsequent measurements. CONCLUSIONS: TLCO is reduced in heart transplant candidates and declines further after heart transplantation despite improvement in lung volumes and airway function. The early and non-progressive nature of TLCO decline suggests an aetiology exerting its effect on TLCO within the first 6 weeks after transplantation. PMID- 9332930 TI - Successful resection of obstructing airway granulation tissue following lung transplantation using endobronchial laser (Nd:YAG) therapy. AB - OBJECTIVE: Airway obstruction due to an excessive growth of granulation tissue at the level of the anastomosis is an important complication following lung transplantation which requires early diagnosis and treatment. We report encouraging experience in the management of this condition using endobronchial Nd:YAG laser therapy. METHODS: Four adult lung transplant recipients developed airway anastomotic obstruction secondary to granulation tissue formation at 9, 10, 32 and 32 days following bilateral sequential lung transplantation (2 patients), en bloc double lung transplantation (1 patient) and single lung transplantation (1 patient). The diameter of the airways at the level of the anastomoses was reduced by 75, 30, 60, 60, 50 and 90%, respectively. Endobronchial Nd:YAG laser was applied via a fiberoptic bronchoscope introduced through a rigid bronchoscope. The granulation tissue was visualised and resected with photocoagulation with the laser using between 1000-2000 J depending on the amount of tissue present. Necrotic tissue was removed with large forceps. If the obstruction extended to the orifice of a lobar bronchus resection was undertaken in a staged fashion. RESULTS: Airway patency was fully restored at two anastomotic sites, and restored to 90% patency at two and 80 and 75% at one each, respectively. This was associated with a significant improvement in pulmonary function in 3 patients. One patient had a subsequent bougie dilatation of a stenotic area and 2 patients received an endobronchial stent for tracheo or broncho-malacia. One patient died from a gastrointestinal haemorrhage. Three patients are well at 10, 17 and 18 months following transplantation and have no further granulation tissue recurrence. There were no complications directly attributable to laser therapy. CONCLUSION: Our encouraging early experience leads us to suggest that Endobronchial Nd-YAG laser therapy should be considered in the management of airway anastomotic obstruction due to excessive granulation tissue formation after lung transplantation. PMID- 9332931 TI - An experimental study of intra aortic balloon pumping within the intact human aorta. AB - OBJECTIVE: Intra-aortic balloon pumping is a therapeutic technique which carries a significant morbidity related to the interaction between the balloon catheter and the aorta. The aim of this study was to visualise directly the dynamic action of the balloon catheter within the cadaveric human aorta in an artificial circulation. METHODS: An artificial circulation was constructed using of PVC tubing, a filter and a roller pump. A series of five intact cadaveric human aortas were then individually studied by placing each in series within the circuit. A balloon catheter was advanced via the left common iliac artery into the descending aorta under direct angioscopic vision. Balloon pumping was then commenced. The circuit was perfused with normal saline at a flow rate of 3 l/min. Pump actions of 1:1 and 1:2 were simulated. Each aorta at the end of the experiment was subjected to histological examination. RESULTS: The balloon only appeared to make direct contact with the wall of the aorta during deflation when it was swept to one side by the circulating fluid. During maximal inflation the only points of contact were the tip of the catheter and the entry site. Side branches of the aorta were not occluded by the balloon. There was considerable atheromatous debris visualised within the lumen of the aorta. Atheromatous plaques were seen to fissure and disrupt by a pressure wave action and not by direct contact with the balloon. CONCLUSION: The balloon catheter moves relative to the wall of the aorta during inflation and deflation. Contact between the balloon and the aorta only occurs during deflation. Side branches of the aorta are not occluded by the catheter. Plaque disruption and embolus formation appear to result from pressure wave action rather than direct contact with the balloon. This may have implications for future balloon design. Further investigation of this poorly understood interaction between the balloon and the aortic wall is required. PMID- 9332932 TI - Descending necrotising mediastinitis--successful treatment using a radical approach. AB - Descending necrotising mediastinitis has been linked with significant morbidity and mortality. We report two recent cases and discuss the surgical management of this serious disease. Complete thoracic drainage was achieved through a median stemotomy, allowing a fast recovery. PMID- 9332933 TI - Remarkable growth of the internal mammary artery used for systemic-to-pulmonary artery shunt in a patient with cyanotic heart disease. AB - The left internal mammary artery-to-the left pulmonary artery shunt was created in a 16-year-old boy with single ventricle, severe pulmonary stenosis palliated by Glenn shunt at the age of two. Four years follow-up angiogram demonstrated a significant increase of the diameter of the left internal mammary artery from 4 to 7 mm. The internal mammary artery is a good alternative conduit for a systemic to-pulmonary artery shunt for a cyanotic heart disease because of its growth potential. PMID- 9332934 TI - Budd-Chiari syndrome as late complication of secundum atrial septal defect closure. AB - A young adult patient, in whom 20 years previously a secundum atrial septal defect had been closed surgically, presented with symptoms of a Budd-Chiari syndrome, cirrhosis of the liver, ascites, and edema of the lower legs. The inferior vena cava-right atrial junction was obstructed by a calcified Teflon patch and shrinkage of the surrounding tissue. Augmentation of the inferior vena cava-right atrial junction with a Gore-Tex patch resulted in unobstructed inflow into the right atrium. PMID- 9332935 TI - Mitral valve anomalies obstructing left ventricular outflow. AB - This paper reports on two cases of more uncommon types of subaortic stenosis. A 2 year-old boy was found with accessory mitral valve leaflet (AMVL) attaching to the anterior leaflet, ballooning into the subaortic ventricular septum associated with a discrete subaortic membrane. The obstruction was successfully relieved by removal of the AMVL and resection of the membrane. A 19-day-old newborn with accessory tissue on the mitral valve (AMVT) causing subaortic stenosis, subaortic ventricular septal defect (VSD) and patent ductus arteriosus was operated on successfully. Accessory tissue excision through the VSD, VSD patch closure and ductus ligation were performed. PMID- 9332936 TI - Migration of pectus excavatum correction bar into the left ventricle. AB - We present the case of a 19-year-old student who underwent correction of a pectus excavatum deformity using a pectus bar. At least 6 months following surgery, one end of the bar had migrated into his right ventricle, across the interventricular septum, to lie with its free end in the left ventricular cavity. This acted as a source of thrombus formation and lead to several systemic embolic events. The patient made a full recovery after removal of the bar. A review of the literature demonstrates that this has not been reported before. PMID- 9332937 TI - Intramural hematoma of the thoracic aorta: precursor sign to thoracic aortic dissection. AB - A 60-year-old man was admitted at his local hospital for persistent chest pain and suspicion of aortic dissection. No evidence of aortic dissection or intimal disruption was noted by means of computed tomography and transesophageal echocardiography. A localized intramural hematoma of the ascending aorta was found. He was first treated medically and remained asymptomatic for 8 days when he developed a new episode of chest pain. He was found to have an acute type A dissection by computed tomography. He underwent graft replacement of the ascending aorta and had an uneventful post-operative course. This case report describes the development of true aortic dissection in a patient who previously had a localized intramural hematoma. PMID- 9332938 TI - Spontaneous resolution late after aortic dissection. AB - A 50-year-old man was operated on for acute type I (DeBakey classification) aortic dissection. The supracoronary ascending aorta was replaced with an interposition graft. Postoperative computed tomography and angiography clearly revealed a double-barrelled aortic arch, left common carotid artery and descending thoracoabdominal aorta with contrast filling of both true and false lumen starting from the distal anastomosis. The same finding was noted at 1 year follow-up with severe compression of the true lumen by the false lumen. At this time, anticoagulation therapy was stopped. One year later, computed tomography showed spontaneous resolution of the dissection in the aortic arch, left common carotid artery and descending aorta over its full length. This was confirmed by angiography. This case reports illustrates that spontaneous resolution of a dissected descending aorta can occur late after surgery from type 1 dissection, but it remains very rare. PMID- 9332939 TI - Complete myocardial revascularization with left heart bypass without oxygenator on the beating heart. AB - BACKGROUND: Whereas the complete myocardial revascularization is necessary in high risk group patients, the CABG procedure on the beating heart on circumflex artery still presents a dilemma. METHODS: Between January 1994 and September 1996, we performed complete myocardial revascularization with left heart bypass in 62 patients (54 male, 8 female, mean age: 57) who had absolute or relative contraindications for CPB. RESULTS: The hospital mortality was 3.2%, late mortality was 1.6%. Peroperative MI was seen in 2 patients (3.2%). The mean number of distal anastomosis was 3.6 (ranged 2-6). CONCLUSION: Complete myocardial revascularization on the beating heart can be performed by using left heart bypass (LHBP) without using an oxygenerator safely in high risk patients. PMID- 9332940 TI - Minimal-invasive, video-assisted vein harvesting for cardiac and vascular surgical procedures. AB - Harvesting of the saphenous vein is a routine procedure in coronary and peripheral vascular surgery. It is usually performed using a continuous long skin incision. Minor complications are reported in up to 24% (hematoma, wound dehiscence, infection, pain) and major problems necessitating surgical interventions (bleeding, abscess) in less than 1%. These complications lead to a prolonged hospital stay. To reduce these complications we have used a new endoscopic, video-assisted technique in 17 patients. Harvesting of the total length of the saphenous vein is possible with only one 2-3 cm long incision proximally the knee joint. We conclude that this technique is safe, may reduce the morbidity of saphenous vein harvesting and is associated with a perfect cosmetic result. PMID- 9332964 TI - The structure of problem and positive behavior among American Indian adolescents: gender and community differences. AB - Using Problem-Behavior Theory as framework, the latent structure of problem and positive behaviors was examined within a sample of 1,894 American Indian adolescents. Support was found for a two-factor second-order structure in which problem behaviors (antisocial behavior, alcohol use, drug use, and risky sexual behavior) and positive behaviors (school success, cultural activities, competencies, and community-mindedness) represented two relatively uncorrelated aspects of behavior. Hierarchical multiple regressions demonstrated that the positive behaviors construct contributed significant incremental construct validity in the statistical prediction of psychosocial outcomes, over and above the problem behaviors. In addition, the fit of the structure was examined across gender and the four participating communities. The importance of the inclusion of positive behaviors is discussed from the standpoint of both prevention/promotion activities and the communities' perceptions. Further recommendations are made for deeper understandings of community concerns and strengths in conducting preventive/promotive research efforts. PMID- 9332965 TI - Adolescent AIDS prevention in context: the impact of peer educator qualities and classroom environments on intervention efficacy. AB - Peer-led, school-based interventions show promise for preventing AIDS among adolescents, but little is known about the processes underlying effective peer education or the conditions that promote its efficacy. This study examined the implementation in one school of an effective, peer-led AIDS prevention program for inner-city 7th-grade participants (n = 123) and identified the qualities of peer educators (n = 15) and classroom environments (n = 5) that contributed to improvement in participants' postintervention AIDS-related attitudes. The Peer Educator Rating Scale was developed to assess two dimensions of participants' perceptions of peer educators: "positive regard" and "perceived similarity." Participants reported greater positive regard for more highly individuated and less shy peer educators, and participants' positive regard for peer educators in turn was associated with lowered AIDS risk as measured by perceptions of peer norms regarding sexual activity and self-efficacy for peer communication regarding sexual topics and condoms. Participants' perceived similarity was not associated with any postintervention improvements in AIDS-related knowledge and attitudes. Participants' classroom membership was associated with improvements in all 5 knowledge and attitude scales, and exploratory classroom-level findings indicated that classroom intervention environments perceived as more organized by participants showed slightly greater overall improvements across AIDS-related knowledge and attitudes scales. Consistent with individual-level findings regarding participants' positive regard for peer educators, the two classrooms with the greatest positive regard for their peer educator teams showed the most student improvement. Implications for further research and the design of future prevention and promotion programs for adolescents are discussed. PMID- 9332966 TI - Mood management mail intervention increases abstinence rates for Spanish-speaking Latino smokers. AB - A self-administered mood management intervention program for smoking cessation provided through the mail to Spanish-speaking Latinos resulted in a 23% abstinence rate at 3 months compared to an 11% abstinence rate for a smoking cessation guide alone. Participants (N = 136) were randomly assigned to receive either the cessation guide (the Guia), or the Guia plus a mood management intervention (Tomando Control de su Vida) presented in writing and in audiotape format. At 3 months after random assignment, 16 out of 71 of those assigned to the Guia-plus-mood management condition reported being abstinent (not smoking for at least 7 days) compared to 7 out of 65 in the Guia-only condition (z = 1.8; p = .04, one-tailed). Moreover, those with a history of major depressive episodes, but not currently depressed, reported an even higher abstinence rate in the Guia plus-mood management condition, compared to the Guia-only condition (31 vs. 11%, z = 1.8, p = .04, one-tailed). We conclude that the mood management mail intervention substantially increases abstinence rates, especially for those with a history of major depressive episodes. PMID- 9332967 TI - Homeless youths and young adults in Los Angeles: prevalence of mental health problems and the relationship between mental health and substance abuse disorders. AB - Although understanding of the subsistence patterns, service utilization, and HIV risk behaviors of homeless youths and young adults in increasing, relatively little is known about the epidemiology of mental health problems in this group or the relationships between mental health problems and substance use. This study measured symptoms of depression, low self-esteem, ADHD, suicidality, self injurious behavior (SIB), and drug and alcohol use disorder in a sample of homeless youth and young adults living in Hollywood, CA. Results indicated extremely high prevalences of mental health problems as compared with corresponding rates of mental health problems found among housed youths in previous studies. Prevalence of mental health problems differed by age and ethnicity. African Americans were at lower risk of suicidal thoughts and SIB than were those of other ethnicities. Older respondents and females were at increased risk of depressive symptoms, and younger respondents were at increased risk of SIB. Previous history of sexual abuse and/or assault was associated with increased risk of suicidality and SIB. Risk factors for drug abuse disorders included ethnicity other than African American, homelessness for 1 year or more, suicidality, SIB, depressive symptoms, and low self-esteem. Risk factors for alcohol abuse disorder included male gender, white ethnicity, homelessness for 1 year or more, suicidality, and SIB. Extremely high rates of mental health problems and substance abuse disorders in this sample suggest the need for street based and nontraditional mental health services targeted toward these youths and young adults. PMID- 9332968 TI - Illness-related support and negative network interactions: effects on HIV infected men's depressive symptomatology. AB - Data collected as part of a psychosocial study of gay and bisexual men's experiences of living with HIV infection as a chronic illness were examined to investigate the psychological impact of the perceived availability of illness related support and negative illness-related network interactions in a sample of men from this population. The sample was comprised of 144 HIV-infected non Hispanic white, African American, and Puerto Rican Men living in the New York City metropolitan area. Analyses found evidence of a conjoint (interactive) effect between perceived support and negative network interactions. There was no evidence of either perceived availability of illness-related network support buffering or negative illness-related network interactions amplifying the effect of HIV/AIDS-related physical symptomatology on depressive symptomatology. PMID- 9332969 TI - Phage presentation and affinity selection of a deletion mutant of human interleukin-3. AB - A deletion derivative of the cytokine human interleukin-3 (hIL-3(15-125), comprising amino acids 15-125 of the native protein) was produced as a fusion to the filamentous phage surface protein pIII. The cytokine was detected in association with phage particles by protein immunoblotting. Compared to an equivalent quantity of soluble-cytokine, phage-presented hIL-3(15-125) exhibited reduced biological activity in a hIL-3-dependent cell proliferation assay. The reduction in activity was attributable to presence of phage particles in the assay, rather than directly owing to physical incorporation of the cytokine into the phage particle. Owing to the position of the amber codon in the phagemid vector, the phagemid-produced free hIL-3(15-125) species (designated hIL-3(15 125) epsilon) had 20 amino acids appended to its C-terminus; hIL-3(15-125) epsilon did not exhibit reduced bioactivity. hIL-3(15-125)-presenting phage were affinity-selected with either a hIL-3-reactive polyclonal antibody or with cells expressing the heterodimeric hIL-3 receptor. These data are consistent with the use of phage-display technology for the affinity selection of hIL-3 variants with modified biological properties. PMID- 9332970 TI - Daughterless is required for the expression of cell cycle genes in peripheral nervous system precursors of Drosophila embryos. AB - The function of the neuronal differentiation gene daughterless (da) is required for the proper initiation of neuronal lineage development in all PNS lineages following the selection of neuronal precursor cells. Previous studies have shown that the ubiquitously expressed da protein is required for the proper expression of neuronal precursor genes and lineage identity genes in the PNS of Drosophila melanogaster embryos. These genes are required for differentiation and cell fate determination in the developing PNS. These findings, however, did not explain the failure of the nascent PNS precursors to undergo a normal cell cycle and divide. Here we show that four genes whose products are required for various stages of the cell cycle are misexpressed in the PNS of da mutant embryos. This suggests that all aspects of PNS precursor differentiation examined so far are under the transcriptional control of da. PMID- 9332971 TI - hsp47 and hsp70 gene expression is differentially regulated in a stress- and tissue-specific manner in zebrafish embryos. AB - We have examined differences in the spatial and temporal regulation of stress induced hsp47 and hsp70 gene expression following exposure of zebrafish embryos to heat shock or ethanol. Using Northern blot analysis, we found that levels of hsp47 and hsp70 mRNA were dramatically elevated during heat shock in 2-day-old embryos. In contrast, ethanol exposure resulted in strong upregulation of the hsp47 gene whereas hsp70 mRNA levels increased only slightly following the same treatment. Whole-mount in situ hybridization analysis revealed that hsp47 mRNA was expressed predominantly in precartilagenous cells, as well as several other connective tissue cell populations within the embryo following exposure to either stress. hsp70 mRNA displayed a very different cell-specific distribution. For example, neither stress induced hsp70 mRNA accumulation in precartilagenous cells. However, high levels of hsp70 mRNA were detectable in epithelial cells of the developing epidermis following exposure to heat shock, but not to ethanol. These cells did not express the hsp47 gene following exposure to either of these stresses. The results suggest the presence of different inducible regulatory mechanisms for these genes which operate in a cell- and stress-specific manner in zebrafish embryos. PMID- 9332972 TI - Effects of cyclin A2 noncoding regions on reporter gene translation during early development of Xenopus laevis. AB - The repression of translation of Xenopus cyclin A2 transcripts during early development was examined by analyzing the effects of cyclin A2 noncoding regions using a CAT reporter system. On their own, the 5' and 3' UTRs (untranslated regions) were unable to inhibit reporter translation until approximately the time of the midblastula transition. Transcripts containing the 3' UTR were polyadenylated after fertilization and the midblastula transition. When both noncoding regions flanked a CAT reporter gene, translation was repressed at all stages of development examined in spite of their polyadenylation after fertilization. From these data, we conclude that the 5' and 3' UTRs interact synergically to prevent translation during early development and that the poly(A) tail is insufficient to promote their translation. PMID- 9332973 TI - Bovine parthenogenesis is characterized by abnormal chromosomal complements: implications for maternal and paternal co-dependence during early bovine development. AB - The present study was conducted to examine the karyotypes of parthenogenetic bovine embryos arising from the application of standard oocyte activation and diploidization methods. Bovine cumulusoocyte complexes were collected and matured in vitro for 24 hr prior to oocyte activation with either 5 microM ionomycin or 7% ethanol for 5 min. Groups of activated oocytes were further treated with 5 micrograms/ml cytochalasin D or 1.9 mM 6-dimethylaminopurine (DMAP) for 6 hr. Cleavage varied significantly (P < .05) among the treatment groups with 68.0% of the ethanol- and DMAP-treated oocytes dividing. Blastocyst development did not vary with 18.4 +/- 2.5% of all treated oocytes progressing to this stage. Blastocyst development did not occur in groups subjected to oocyte activation alone. Blastocysts displayed haploid (2.3%), diploid (11.4%), tetraploid (40.9%), octaploid (4.5%), and mixoploid chromosomal complements (40.9%). Two-cell stage parthenogenotes resulting from ethanol or ionomycin treatment alone displayed haploid (66.7%), diploid (16.7%), tetraploid (4.2%), and mixoploid (12.5%) complements. Our results demonstrate that diploid bovine parthenogenotes arising from these procedures are a minority, with the majority of parthenogenotes displaying polyploid and mixoploid chromosomal complements. The events contributing to these abnormal chromosomal complements occur as early as completion of the first cell cycle, possibly linking these events with the absence of a paternally supplied centrosome. PMID- 9332974 TI - Mutagenesis screens for interacting genes reveal three roles for fat facets during Drosophila eye development. AB - The Drosophila fat facets gene encodes a deubiquitinating enzyme that regulates a cell communication pathway essential early during eye development to inhibit the determination of excess photoreceptors. Ubiquitin is a small polypeptide that tags proteins for degradation by a multisubunit proteolytic complex called the proteasome. The FAT FACETS protein is thought to be required to remove ubiquitin from a particular protein, thereby rescuing if from proteolysis. In order to identify the genes encoding the substrate of FAT FACETS and other components of the neural inhibition pathway, a mutagenesis screen for dominant enhancers of the fat facets mutant eye phenotype was performed. Several genes were identified, one of which is an excellent candidate for encoding a component of the pathway regulated by FAT FACETS. Three different eye phenotypes were observed when the fat facets mutants were dominantly enhanced by different mutations, suggesting that fat facets has other functions in addition to its critical role early in eye development. PMID- 9332977 TI - Helminth ectoparasites of sillaginid fishes (Perciformes: Percoidei) have low species richness. AB - Just nineteen species of ectoparasitic helminths were found in a survey of over 1,500 individuals of 26 species of sillaginid fishes in the Indo-west Pacific. A twentieth worm is known only from the literature; a twenty-first, also known only from the literature, is considered a doubtful record. Fifteen of the twenty worms are branchial monogeneans, one is a monogenean of the pharyngeal plates, one is an ectoparasitic digenean living under the scales, and three are leeches of the mouth cavity and fins. The most common monogeneans were diplectanids (Diplectanum spp. and Monoplectanum spp.) and microcotylids (Polylabris spp.), each with five recently described or redescribed species. Of the remaining monogeneans, three were extremely rare, and two were uncommon. Pseudobivagina sp. and Polynemicola sp. (Microcotylidae) and Pseudempleurosoma sp. (Ancyrocephalidae) were represented by only a single worm each from three different hosts (Sillago robusta, S. sihama, and S. ingenuua, respectively). The gyrodactylid Gyrodactylus sp. is widespread and was recorded from four species of sillaginids (S. ciliata, S. japonica, S. schomburgkii and S. sihama). Encotyllabe chironemi Robinson (Capsalidae) is recorded for the first time from sillaginids, but only on S. aeolus. Two additional monogeneans are known from sillaginids only in the literature: Dactylogyrus sp. (Dactylogyridae) is known only from cultured S. sihama; the single specimen of Microcotyle sp. (Microcotylidae) recorded from Sillaginodes punctata is probably a contaminant, since the haptor was missing. The generalist trematode Transversotrema licinum Manter (Transversotrematidae) was found for the first time in samples of four species of sillaginids (Sillago analis, S. ingenuua, S. lutea and S. sihama). Three species of piscicolid leeches were encountered: Austrobdella translucens Badham was common on the fins of three large inshore sillaginids (S. ciliata, S. schomburgkii and S. analis); a single specimen of the generalist species Zeylanicobdella arugamensis De Silva was recovered from S. soringa; and specimens of Z. stellata (Moore) infected S. schomburgkii and S. analis. The diversity of host-specific worms in Sillaginidae is low compared with those of some other Indo-west Pacific fishes. PMID- 9332979 TI - New observations on seuratoid nematodes parasitic in fishes of the Parana River, Brasil. AB - The present paper comprises a systematic survey of nematodes of the superfamily Seuratoidea collected from fishes of the Parana River, southern Brazil, in 1992 1995. The following species were recorded: Neoparaseuratum travassosi Moravec, Kohn et Fernandes, 1992, Spectatus sp., Seuratoidea gen. sp. larvae, Cucullanus brevispiculus Moravec, Kohn et Fernandes, 1993, C. pinnai pinnai Travassos, Artigas et Pereira, 1928, C. pinnai pterodorasi subsp. n., C. rhamphichthydis sp. n., C. zungaro Vaz et Pereira, 1934, and Dichelyne pimelodi sp. n. C. pinnai pterodorasi (type host Pterodoras granulosus) differs from C. p. pinnai in the shape of the oesophagus and in the position of the nerve ring. C. rhamphichthydis (only females) (type host Rhamphichthys rostratus) is characterized mainly by the postoesophageal position of the excretory pore and deirids, complex structure of the cuticular lining of oesophastome and by markedly narrow body, whereas D. pimelodi (one male) (type host Pimelodus sp.) by the length of spicules (1.46 mm), postoesophageal position of the excretory pore and deirids and by the arrangement of genital papillae. Most species are briefly described (females of C. brevispiculus and Seuratoidea gen. sp. larvae for the first time) and illustrated and some problems concerning their taxonomy, hosts and geographical distribution are discussed. PMID- 9332978 TI - Complement fixing antibody production in thymectomized Oncorhynchus mykiss, vaccinated against or infected with the pathogenic haemoflagellate Cryptobia salmositica. AB - Short-term thymectomized (two months after thymectomy) adult rainbow trout, Oncorhynchus mykiss (Walbaum) infected with the flagellate Cryptobia salmositica Katz, 1951 responded well during primary infection with C. salmositica and recovered fish also showed secondary response (rapid production of complement fixing antibody after homologous challenge) five months after recovery. Complement fixing antibody was detected during primary and secondary responses and the titres of complement fixing antibody in thymectomized fish were significantly lower than those in infected intact fish. The parasitaemia in thymectomized infected fish was significantly lower than in intact infected fish. Both recovered thymectomized fish and intact fish were protected from cryptobiosis when they were challenged. Similarly, long-term thymectomized fish (nine months after thymectomy) vaccinated with an attenuated strain of C. salmositica were protected from cryptobiosis. There were no significant difference (P > 0.05) in parasitaemia, packed cell volume and complement fixing antibody titres between vaccinated/challenged thymectomized and vaccinated/challenged intact fish. Hence, thymectomy in adult rainbow trout did not decrease the detectable complement fixing antibody against C. salmositica in long-term thymectomized fish but reduced the detectable protective antibody in short-term thymectomized fish. PMID- 9332980 TI - Occurrence of cystacanths of Polyacanthorhynchus kenyensis larvae (Acanthocephala) in four teleostean fishes from a tropical lake, Lake Naivasha, Kenya. AB - From January 1992 to December 1993, a total of 2158 fish, namely Oreochromis leucostictus (Trewavas, 1983), Micropterus salmoides (Lacepede, 1802), Tilapia zillii (Gervais, 1848) and Barbus amphigrama (Boulenger, 1902) were sampled from thirteen stations on Lake Naivasha, Kenya, using a fleet of gill nets and examined for helminth parasites. The prevalence of infection due to cystacanths of an acanthocephalan, Polyacanthorhynchus kenyensis Schmidt et Canaris, 1967 among parasitized O. leucostictus ranged from 30.4 to 86.9%; among T. zillii from 4.1 to 77.7%; in M. salmoides from 20 to 50%; and in B. amphigrama from 5.8 to 100%. In 735 hosts belonging to the above four species, a total of 4198 immature specimens of P. kenyensis were recovered. All cystacanths were found in extraintestinal sites, either free within the fish body cavity or encysted within the host visceral organs. There was no significant variation in the prevalence of the parasite within months (P > 0.001). Host sex ratio was significant (P < 0.001) in favour of male T. zillii, and also highly significant (P < 0.001) in favour of male O. leucostictus. Moreover, in this fish, prevalence of infection was observed to increase with the increase in the size of the fish. Among infected M. salmoides, there was no significant departure from a 1:1 sex ratio. PMID- 9332981 TI - Two gynandromorphs of Ixodes (Ixodes) Pacificus (Acari: Ixodidae) from California, USA. PMID- 9332982 TI - Anergy skin testing and tuberculosis [corrected] preventive therapy for HIV infected persons: revised recommendations. Centers for Disease Control and Prevention. AB - This report updates and supersedes previous recommendations (MMWR 1991;40[No. RR 5]:27-33) for the use of anergy skin testing in conjunction with purified protein derivative (PPD)-tuberculin skin testing of persons infected with human immunodeficiency virus (HIV). In February 1997, CDC convened a meeting of consultants to discuss current information regarding anergy skin testing, PPD skin testing, and tuberculosis (TB) preventive therapy for HIV-infected persons. In formulating these recommendations, CDC considered the results of this meeting, as well as a review of published studies pertaining to PPD and anergy skin testing of persons who are infected with HIV. Isoniazid preventive therapy is effective in reducing the incidence of active TB among persons who have HIV infection and latent TB. Because of the complications associated with TB disease in HIV-infected persons, these persons must be screened for tuberculin infection. HIV-infected persons who have positive reactions to skin testing with PPD tuberculin should be evaluated to exclude active TB and offered preventive therapy with isoniazid if indicated. However, HIV-infected persons may have compromised ability to react to PPD-tuberculin skin testing, because HIV infection is associated with an elevated risk for cutaneous anergy. Anergy testing is a diagnostic procedure used to obtain information regarding the competence of the cellular immune system. When a clinician elects to use anergy testing as part of a multifactorial assessment of a person's risk for TB, the two Food and Drug Administration-approved Mantoux-method tests (mumps and Candida), used together, with cut-off diameters of 5 mm of induration, are recommended. Efforts to apply the results of anergy testing to preventive therapy decisions must be supplemented with information concerning the person's risk for infection with Mycobacterium tuberculosis. Factors limiting the usefulness of anergy skin testing include problems with standardization and reproducibility, the low risk for TB associated with a diagnosis of anergy, and the lack of apparent benefit of preventive therapy for groups of anergic HIV-infected persons. Therefore, the use of anergy testing in conjunction with PPD testing is no longer recommended routinely for screening programs for M. tuberculosis infection conducted among HIV-infected persons in the United States. PMID- 9332983 TI - Selective oestrogen receptor modulation: HRT replacement therapy? PMID- 9332984 TI - Pregnancy post-transplant: the establishment of a UK registry. PMID- 9332985 TI - Guidelines on the investigation and management of thrombocytopenia in pregnancy and neonatal alloimmune thrombocytopenia. PMID- 9332986 TI - Newer HRT regimens. PMID- 9332987 TI - The identification and long term effects of fetal growth restriction. PMID- 9332989 TI - Computerised estimation of the baseline fetal heart rate in labour: the low frequency line. AB - OBJECTIVE: To develop and evaluate a computerised algorithm for the estimation of the fetal heart rate baseline (low frequency line) during labour. DESIGN: Retrospective observational study. METHODS: Fetal heart rate signals were obtained from women in labour using the Nottingham fetal ECG monitor. The computerised algorithm for the baseline estimation was developed for intrapartum applications and is based on averaging modal fetal heart rate values. Evaluation was carried out on sixty cardiotocographic recordings by 12 experts and by the computer. These estimates were compared with those obtained from the computerised system using paired differences and intraclass correlation. RESULTS: The study showed that it is possible to produce a low frequency line from data obtained from intrapartum records. The system could not estimate the low frequency line in four records, whereas experts were also unable to estimate between one and seven tracings. The 95% CI for the paired differences between computer and experts was 12 to 15 bpm, whereas between the experts this was -10 to 10. With the exception of one expert, there was a high concordance between experts and between computer and experts (intraclass correlation > 0.9). CONCLUSIONS: The performance of this computerised algorithm cannot be distinguished from that of experienced clinicians. There were no significant differences between baseline values obtained by the computerised algorithm and those by the clinicians. PMID- 9332988 TI - Long term outcome after umbilical artery acidaemia at term birth: influence of gender and duration of fetal heart rate abnormalities. AB - OBJECTIVE: To study the outcome after acidaemia at term birth, and the relation to gender and duration of pathological fetal heart rate changes. DESIGN: Population based study of 154 infants with umbilical artery pH < 7.05 at term birth. Neonatal outcome and the result of developmental screening at age four years were compared with a control group with pH > 7.10. Fetal heart rate traces in infants with acidaemia were reviewed, and the relation between duration of fetal heart rate changes and outcome was analysed. RESULTS: Of the 154 newborns with acidaemia at birth, 10 had encephalopathy, of which two died and two developed cerebral palsy. Nine of these 10 infants were boys, and eight had pH < 7.00. Male newborns (n = 39) more often had pronounced acidaemia (pH < 7.00) than females (n = 22). Although few infants had severe impairment, infants born with acidaemia significantly more often had speech problems at follow up than controls (19/102 versus 8/98; P = 0.03). In infants with acidaemia, duration of abnormal fetal heart rate changes was significantly associated with neonatal encephalopathy and speech problems at age four years. CONCLUSIONS: Acidaemia at term birth was associated with neonatal encephalopathy and with speech problems at four years of age. Boys had more often pronounced acidaemia and a complicated course. A protracted abnormal fetal heart rate trace was associated with poor outcome. PMID- 9332990 TI - Amniotic fluid nitric oxide and uteroplacental blood flow in pregnancy complicated by intrauterine growth retardation. AB - OBJECTIVE: To examine the correlation between placental nitric oxide production and uteroplacental blood flow. PARTICIPANTS: Thirty-one pregnant women with fetuses with intrauterine growth retardation and 27 normal pregnancies as controls. DESIGN: Correlation between amniotic fluid measurements of nitrite metabolite in the third trimester and flow velocimetry waveforms recorded from uterine, umbilical and fetal middle cerebral arteries. Intrauterine growth retarded pregnancies were compared with controls. MAIN OUTCOME MEASURES: Concentrations of nitric oxide metabolites (NO2- and NO3-) in amniotic fluid were correlated with flow velocimetry waveforms findings by the determination of correlation coefficient. RESULTS: Overall median nitrite values in amniotic fluid were higher (P < 0.01) in intrauterine growth retarded patients (median 8.6 micromol/mg creatinine) than in controls (5.6 micromol/mg creatinine). Pathologic uterine flow velocimetry waveforms in uterine artery (-2SD) were observed in 12 women of the intrauterine growth retarded group, and the concentration of amniotic fluid nitrite was significantly lower (P < 0.01) in these patients (median 4.45 micromol/mg creatinine) than in those with normal flow velocity waveforms (median 11.43 micromol/mg creatinine). A significant negative correlation was observed between nitrite concentrations and uterine artery resistance index, umbilical artery pulsatility index and umbilical artery pulsatility index:middle cerebral artery pulsatility index ratio. CONCLUSIONS: We conclude that placental nitric oxide is significantly associated with uteroplacental blood flow and may be important in maintaining adequate uteroplacental perfusion in intrauterine growth retarded pregnancies. PMID- 9332991 TI - Lipid peroxidation in cord blood: the effect of amniotic fluid volume. AB - OBJECTIVE: To assess the effect of amniotic fluid volume on umbilical cord arterial lipid peroxide levels in relation to intrapartum events. DESIGN: Prospective observational study. SETTING: Delivery suite of a teaching hospital, the Chinese University of Hong Kong. POPULATION: Women with singleton, term, cephalic presentation, and an initially normal fetal heart rate tracing. METHODS: All pregnancies had amniotic fluid index assessments before and after amniotomy and cord arterial lipid peroxide determination at delivery. Multiple regression analysis. MAIN OUTCOME MEASURES: Cord arterial malondialdehyde and organic hydroperoxide levels. RESULTS: In 247 cases following amniotomy levels were inversely correlated with intrapartum amniotic fluid index. Amniotic fluid index during labour was an independent determinant of cord arterial lipid peroxide concentration, along with duration of second stage, absence of epidural, presence of tight nuchal cord entanglement and evidence of fetal distress. CONCLUSIONS: Oligohydramnios during labour is associated with high levels of lipid peroxidation in the fetus, reflecting an increase in hypoxic cellular damage by free radicals. PMID- 9332992 TI - Lipid peroxidation in cord blood: a randomised sequential pairs study of prophylactic saline amnioinfusion for intrapartum oligohydramnios. AB - OBJECTIVE: To determine the efficacy of prophylactic intrapartum amnioinfusion in reducing cord arterial lipid peroxide levels in cases of intrapartum oligohydramnios. DESIGN: Sequential randomised pairs trial. SETTING: Delivery suite of a teaching hospital, the Chinese University of Hong Kong. POPULATION: Women with singleton, term pregnancy, cephalic presentation, clear amniotic fluid and an amniotic fluid index < or = 5 cm, with a normal intrapartum fetal heart rate tracing within 30 minutes of amniotomy. METHODS: Selected patients were randomised either for prophylactic saline amnioinfusion or as control cases. Cord arterial lipid peroxide concentrations and acid base balance were determined at delivery. MAIN OUTCOME MEASURES: Operative intervention for fetal distress, cord arterial malondialdehyde and organic hydroperoxide levels, pH and base excess. RESULTS: Amnioinfusion was associated with significant reductions in the incidence of operative delivery for fetal distress and in lipid peroxide levels, an increase in base excess, but no significant alteration in pH. CONCLUSIONS: Oligohydramnios in labour is associated with high levels of lipid peroxidation, reflecting cellular damage by release of free radicals following hypoxia reperfusion. Prophylactic intrapartum saline amnioinfusion is an effective technique for the reduction of lipid peroxidation and of the incidence of operative intervention for fetal distress but has no significant effect on overall operative delivery rates. PMID- 9332993 TI - Loss of endothelium-dependent relaxation in myometrial resistance arteries in pre eclampsia. AB - OBJECTIVE: To measure endothelium-dependent relaxation in myometrial resistance arteries and to compare this parameter in nonpregnant and normotensive pregnant women and those with pregnancies complicated by pre-eclampsia. DESIGN: A prospective study. SETTING: Nottingham City Hospital. SAMPLE: Thirty-seven nonpregnant women undergoing hysterectomy and 51 pregnant women undergoing caesarean section, of whom there were 39 normotensive and 12 with pre-eclampsia. METHODS: Resistance arteries, dissected from myometrial biopsies, were mounted on a wire myograph and preconstricted with vasopressin then subjected to incremental doses of bradykinin. RESULTS: Endothelium-dependent relaxation to bradykinin was seen in the vessels of nonpregnant and normotensive pregnant women. Markedly reduced endothelium-dependent relaxation was found in the myometrial arteries from women with pre-eclampsia when compared with both nonpregnant (P < 0.0001) and normotensive pregnant (P < 0.0001) women. CONCLUSION: A significant loss of endothelium-dependent relaxation in myometrial resistance arteries in pre eclampsia may contribute to the altered vasoreactivity seen in pre-eclampsia, and particularly to the decreased placental perfusion and fetal growth retardation commonly associated with this condition. PMID- 9332994 TI - Improved methods of assessing proteinuria in hypertensive pregnancy. AB - OBJECTIVE: To determine whether use of an automated urinalysis device will improve the accuracy of detecting proteinuria, and whether spot urine protein to creatinine ratio will provide accurate quantitation of proteinuria in hypertensive pregnant women. DESIGN: Prospective studies assessing the accuracy of both detection and quantitation of proteinuria. SETTING: Antenatal ward and pregnancy day assessment unit of St George Hospital, a teaching hospital in Sydney, Australia. POPULATION: Hypertensive pregnant women admitted to hospital or day assessment unit for management of their hypertensive disorders. METHODS: 1. Routine dipstick urinalysis and 2. urinalysis by an automated device (Clinitek 100 Ames) on a midstream urine sample were compared with measurement of protein concentration on that sample (n = 103). In a third study, the protein:creatinine ratio on a midstream (spot) urine sample was compared with protein excretion over the subsequent 24 hours (n = 100). MAIN OUTCOME MEASURES: Relations between urine protein concentrations and 1. dipstick urinalysis and 2. automated urinalysis; 3. Positive and negative predictive values of spot protein:creatinine ratio for true proteinuria (> or = 300 mg/day). RESULTS: Automated urinalysis improved the percentage of true positive urinalyses from 48% with visual urinalysis to 74% (P = 0.02). True negatives were 98% to 100% for both methods. Spot urine protein:creatinine ratio correlated well with subsequent 24-hour urine proteinuria (r = 0.93, P < 0.001). A protein:creatinine ratio > 30 mg protein/mmol creatinine was the optimum discriminant value for true proteinuria, with sensitivity 93%, specificity 92%, positive predictive value 95% and negative predictive value 90%. CONCLUSIONS: Use of an automated urinalysis device improved accurate detection of proteinuria, particularly reducing false positive tests. A random urine protein:creatinine ratio provides an accurate and rapid quantitation of proteinuria in hypertensive pregnant women. This should improve clinical care, especially when managing hypertensive pregnant women as outpatients. PMID- 9332995 TI - Single photon emission and cerebral computerised tomographic scan and transcranial Doppler sonographic findings in eclampsia. AB - OBJECTIVE: To define more clearly the neuropathophysiology of eclampsia utilising single-photon emission computed tomography (SPECT), cerebral computerised tomography (CT) and transcranial Doppler (TCD) ultrasonography. DESIGN: A prospective study SETTING: The obstetric unit in King Edward VIII Hospital, a large tertiary referral centre in Kwa-Zulu Natal, South Africa. PARTICIPANTS: Sixty-five women with eclampsia. INTERVENTIONS: Imaging and ultrasonographic investigations were performed within 48 hours postpartum. Unenhanced cerebral CT scans were performed in all the women and SPECT scans were performed on 63 women using Technetium-99m-hexamethylpropyleneamineoxime (99mTc-HMPAO) as a tracer of regional cerebral blood flow. Middle cerebral artery blood flow velocity waveforms were measured using 2 MHz pulsed Doppler ultrasound via the transtemporal approach. MAIN OUTCOME MEASURES: Abnormalities in SPECT scan, CT scan and TCD ultrasound findings. RESULTS: SPECT scanning revealed perfusion deficits in the watershed areas in all women, 75% of whom had concomitant deficits in the parieto-occipital areas of the brain. Hypodensities (cerebral oedema) were reported in 38 CT scans (58.5%), with parieto-occipital involvement in 97.4% of cases. Increased flow velocity measurements in the middle and posterior cerebral arteries were recorded in 36 (85.7%) women in whom TCD ultrasound was performed. CONCLUSION: The pathophysiological mechanism of eclamptic seizures is primary cerebral vasospasm with resultant ischaemia and cerebral oedema involving mainly the watershed areas and parieto-occipital lobes of the brain. SPECT scanning has been shown to be superior to CT scanning and TCD ultrasonography in detecting neuropathophysiologic alterations in eclampsia. However, each of the three investigative tools provide its own unique information and all three are necessary research techniques to improve our understanding of the neuropathophysiological mechanism of eclamptic seizures. PMID- 9332996 TI - Seizure prophylaxis in hypertensive pregnancies: a framework for making clinical decisions. AB - OBJECTIVE: To describe a framework for generating therapeutic recommendations using seizure prophylaxis in hypertensive pregnancies as an example. DESIGN: A decision-making framework was built using: 1. evidence of therapeutic benefit, with number needed to treat as the effect measure; 2. baseline rates of the target disorder that the treatment was designed to prevent; and 3. a treatment threshold, determined by weighting the potential risks against the potential benefits of the treatment. METHODS: Evidence of therapeutic benefit (i.e. reduction in eclamptic seizures associated with magnesium sulphate therapy in hypertensive pregnancies) was determined by a systematic quantitative overview of controlled clinical trials. Baseline rates of seizures without magnesium sulphate therapy were derived from a recent cohort study. A treatment threshold was generated using estimates of treatment associated morbidities which were weighted against the potential reduction in seizures from magnesium sulphate therapy considering the relative values assigned to these outcomes by obstetricians practising in our hospital. RESULTS: The number of hypertensive women needed to be treated with magnesium sulphate to prevent a single case of eclamptic seizures varied in a curvilinear fashion dropping from 1000 to 14 as the baseline rate of seizures increased from 0.1% to 10%. The treatment threshold as measured by number needed to treat was 64 (range 57-77). The number needed to treat for nonproteinuric hypertension was 1000 (95% CI 180-40,000), whereas it was 32 (95% CI 20-57) for proteinuric hypertension. Considering the uncertainty in estimation of the numbers needed to treat and treatment threshold, magnesium sulphate therapy may be recommended for women at high risk of eclampsia (e.g. severe pre eclampsia) while it should be withheld in cases at low risk (e.g. nonproteinuric hypertension and mild pre-eclampsia). CONCLUSION: While awaiting further research obstetricians intuitively make decisions about seizure prophylaxis in hypertensive pregnancies. Our decision-making framework generated therapeutic recommendations by explicit consideration of the available evidence. PMID- 9332997 TI - Low dose dopamine in postpartum pre-eclamptic women with oliguria: a double blind, placebo controlled, randomised trial. AB - OBJECTIVE: To assess the effect of low dose dopamine on the urine output in postpartum pre-eclamptic or eclamptic women with oliguria. DESIGN: A double blind, randomised controlled study. SETTING: The high care area of the labour ward in a teaching hospital. SAMPLE: Forty postpartum pre-eclamptic women with oliguria, defined as < 30 mL/hour, who have not responded to a 300 mL crystalloid fluid challenge. INTERVENTION: Dopamine was infused at a rate of 1 to 5 microg/kg per minute, or sterile water was given as placebo in the same dilution. MAIN OUTCOME MEASURE: Urine output, blood pressure and pulse was measured for six hours before and for six hours after the intervention. RESULTS: Women who received dopamine (344 mL over 6 hours) showed a clinically and statistically significant (P = 0.0014, Mann-Whitney U test) higher median urine output compared with those receiving placebo (135 mL over 6 hours) for the duration of therapy. The respective 95% confidence intervals were 212.3 to 712.7 mL compared with 73.8 to 244.7 mL. No differences in blood pressure or pulse were found between the two groups. CONCLUSIONS: The use of low dose dopamine in a labour setting improved urine output in postpartum pre-eclamptic women with oliguria who had not responded to a single fluid challenge without a detrimental effect on the blood pressure or pulse. PMID- 9332998 TI - Continuous transdermal oestrogen and interrupted progestogen as a novel bleed free regimen of hormone replacement therapy for postmenopausal women. AB - OBJECTIVE: To investigate the effects of a hormone replacement therapy regime of continuous oestrogen and interrupted progestogen, administered transdermally, on the endometrium of postmenopausal women, the pattern of bleeding and relief of menopausal symptoms. DESIGN: Volunteer pilot study of up to six months duration involving weekly application of an oestrogen-only skin patch releasing 50 microg oestradiol per day interspersed with a combined oestrogen and progestogen patch releasing 50 microg oestradiol and 250 microg norethisterone acetate per day for three days. Transvaginal ultrasound measurements of endometrial thickness and endometrial biopsies were performed in the third month of treatment at the end of both an oestrogen-only phase of treatment and a combined oestrogen-progestogen phase. SETTING: Specialist community menopause clinic, Dean Terrace Centre, Edinburgh. PARTICIPANTS: Fifteen healthy postmenopausal women. MAIN OUTCOME MEASURES: Effect of treatment on endometrial histology, the immunolocalisation of oestrogen and progesterone receptors and the cell proliferation marker Ki 67 after three months of treatment and the proportion of women without bleeding at six months. RESULTS: Treatment provided relief of hot flushes and by the sixth month of study 10 of the 14 women who completed treatment had no vaginal bleeding (71%). No endometrial hyperplasia or atypical changes were observed in biopsies and ultrasound measurements of endometrial thickness demonstrated a thin endometrium. Reduced immunostaining for Ki 67 was observed in endometrium from the combined phase of treatment compared with the oestrogen-only phase, consistent with a progestogenic-antagonism of proliferation. Exposure to progestogen did not suppress steroid receptors as similar immunostaining was observed in both treatment phases. CONCLUSIONS: Continuous oestrogen and interrupted progestogen administered transdermally offers promise as a novel bleed-free hormone replacement therapy for postmenopausal women. PMID- 9333000 TI - Abdominal radical trachelectomy: a new surgical technique for the conservative management of cervical carcinoma. AB - Traditionally radical hysterectomy has formed the mainstay of treatment for early stage cervical carcinoma. More recently radical trachelectomy and laparoscopic lymphadenectomy have been introduced to allow preservation of fertility. We present a new approach to fertility-sparing surgery, namely abdominal radical trachelectomy. The technique is similar to a standard radical hysterectomy and lymphadenectomy. In our technique the ovarian vessels are not ligated and, following lymphadenectomy and skeletonisation of the uterine arteries, the cervix, parametrium and vaginal cuff are excised. The residuum of the cervix is then sutured to the vagina and the uterine ateries re-anastomosed. PMID- 9332999 TI - The impact of hormone replacement therapy on quality of life and willingness to pay. AB - OBJECTIVE: To measure the gain in quality of life due to hormone replacement therapy for women with mild and severe menopausal symptoms. DESIGN: Prospective study where data on quality of life and willingness to pay were collected by interview. SETTING: Department of Gynaecology at Sodertalje Hospital near Stockholm. PARTICIPANTS: One hundred and four women aged 45 to 65 years treated for menopausal symptoms for at least one month. METHODS: Quality of life was measured by the time tradeoff and rating scale methods. The willingness to pay for hormone replacement therapy was investigated using the contingent valuation method. MAIN OUTCOME MEASURES: The quality adjusted life year weight measured with the rating scale and time tradeoff methods, and willingness to pay. RESULTS: The increase in the quality adjusted life year weight due to hormone replacement therapy for women with mild symptoms was 0.26 according to the rating scale method and 0.18 according to the time tradeoff method. For women with severe symptoms the quality adjusted life year weight increased by 0.50 according to the rating scale method and by 0.42 according to the time tradeoff method. The mean willingness to pay for hormone replacement therapy per month was 2300 Swedish krone for women with mild symptoms and 4800 Swedish krone for women with severe symptoms (Pounds 1 = 10.3 Swedish krone). CONCLUSIONS: Hormone replacement therapy leads to a major improvement in quality of life for women with menopausal symptoms. Both for women with mild and severe menopausal symptoms the willingness to pay for the treatment also greatly exceeds the costs, indicating that hormone replacement therapy is economically beneficial for women with menopausal symptoms. PMID- 9333001 TI - Effects of embryo reduction from trichorionic triplets to twins. AB - Sixty-six trichorionic triplet pregnancies reduced to twins were compared with 47 triplet pregnancies that were not reduced. The miscarriage rate was higher (7.6% compared with 2.6%) but the number delivering between 24 and 32 weeks was lower (8.2% compared with 24.0%). Since severe preterm delivery is associated with risks of neonatal death and severe handicap, embryo reduction of triplets to twins may not improve the chance of survival but may reduce the rate of handicap. PMID- 9333002 TI - The hidden mortality of monochorionic twin pregnancies. AB - In an ultrasound screening study at 10 to 14 weeks of gestation for measurement of fetal nuchal translucency thickness there were 102 monochorionic and 365 dichorionic twin pregnancies. In the monochorionic compared with the dichorionic pregnancies there was a higher rate of fetal loss before 24 weeks of gestation (12.2% versus 1.8%), perinatal mortality (2.8% versus 1.6%), prevalence of delivery before 32 weeks (9.2% versus 5.5%), and prevalence of birthweight below the 5th centile in both twins (7.5% versus 1.7%). However, the proportion of pregnancies with a birthweight discordancy of more than 25% was similar in the two groups (11.3% versus 12.1%). PMID- 9333003 TI - Fetal heart rate at 10 to 14 weeks and birthweight. AB - In 6644 singleton pregnancies resulting in live births, the fetal heart rate was measured at 10 to 14 weeks of gestation (median 12). There was no significant association between fetal heart rate and birthweight. These findings demonstrate that if there is an association between fetal heart rate, birthweight and subsequent development of cardiovascular disease the responsible intrauterine insult and/or the adaptive fetal response are not present at 10 to 14 weeks of gestation. PMID- 9333004 TI - Detection of glove puncture and skin contamination during caesarean section. AB - Measures that can be taken to reduce exposure to potentially infected body fluids are of particular relevance in obstetric and gynaecological surgery due to high rates of glove puncture and relatively higher prevalence of human immunodeficiency virus seropositivity in the obstetric age group. We describe the use of a simple electronic device that alarms following puncture of surgical gloves or the creation of a fluid bridge between surgeon and patient. Further exposure to potentially infected body fluids is thus prevented. This present study was performed in the context of caesarean section, but the application of the technique to gynaecological procedures is appropriate. PMID- 9333005 TI - HLA-G polymorphism in Finnish couples with recurrent spontaneous miscarriage. AB - HLA-G is a class I major histocompatibility complex gene that is expressed in cytotrophoblasts at the materno-fetal interface. It has been suggested that HLA-G could play a key role in materno-fetal immunological interactions during pregnancy. To investigate whether there is an association between HLA-G locus and recurrent spontaneous miscarriage, HLA-G alleles were determined by a PCR-RFLP method in 38 couples with recurrent spontaneous miscarriage and in 26 random control couples. In this series parental HLA-G sharing, extended HLA-G/A haplotypes and the frequencies of the HLA-G alleles were similar in the two groups. Thus, our data suggest that there is no detectable relation between susceptibility to recurrent spontaneous miscarriage and HLA-G locus. PMID- 9333006 TI - Liver function tests in pre-eclampsia: importance of comparison with a reference range derived for normal pregnancy. PMID- 9333007 TI - Randomised trial comparing a policy of early with selective amniotomy and uncomplicated labour at term. PMID- 9333008 TI - Randomised trial comparing a policy of early with selective amniotomy and uncomplicated labour at term. PMID- 9333009 TI - Successful methotrexate treatment of a viable pregnancy within a thin uterine scar. PMID- 9333010 TI - Ureteric injury following laparoscopic colposuspension. PMID- 9333011 TI - Antioxidants in the treatment of severe pre-eclampsia: an explanatory randomised controlled trial. PMID- 9333012 TI - Vaginal endoscopic oophorectomy with vaginal hysterectomy: a simple minimal access surgical technique. PMID- 9333013 TI - Constitutive membrane association potentiates activation of Bruton tyrosine kinase. AB - Mutations in the nonreceptor tyrosine kinase Btk result in the B cell immunodeficiencies X-linked agammaglobulinemia (XLA) in humans and X-linked immunodeficiency (xid) in mice. Genetic and biochemical evidence implicates Btk as a key component of several B cell signaling pathways. Activation of Btk by a point mutation (E41K) within the PH domain (Btk*) results in fibroblast transformation and is correlated with increased membrane localization of Btk. When wild type Btk is activated by coexpression with Lyn, the tyrosine phosphorylated pool of Btk is highly enriched in the membrane fraction. To determine whether membrane association is sufficient to activate Btk, we targeted Btk to the plasma membrane using a series of fusion proteins including GagBtk, CD16Btk and CD4Btk. Constitutive membrane association greatly enhanced the ability of Btk to transform Rat2 fibroblasts in the presence of high levels of Src activity. All membrane targeted forms of Btk were highly tyrosine phosphorylated. Transformation required membrane localization, Btk kinase activity, transphosphorylation by Src family kinases, and an intact SH2 domain but not the PH or SH3 domains. These data suggest that membrane localization is a critical early step in Btk activation. PMID- 9333015 TI - Butyrate modulates DNA-damage-induced p53 response by induction of p53 independent differentiation and apoptosis. AB - Butyrate, a physiologically occurring agent, has been reported to decrease constitutively high expressed p53 levels in transformed cells. To elucidate whether butyrate also inhibits DNA-damage-induced p53 response we investigated the effects of butyrate and the anticancer drug mitomycin C in normal C3H10T1/2 cells harbouring wild-type p53. In comparison with p53-deficient fibroblasts we examined p53 protein level, cell cycle arrest, differentiation, and apoptosis. Butyrate induced G1 phase arrest, differentiation, and p53-independent increase in p21(waf1/cip1) protein. Moreover, butyrate induced p53-independent apoptosis, which was, as well as p53-mediated apoptosis, associated with a dose-dependent increase in Bax and c-Myc protein. Pretreatment with butyrate repressed dose dependently mitomycin-C-induced p53 accumulation and interfered with p53 dependent cell cycle arrest. Butyrate further partially inhibited p53-mediated apoptosis, but low doses of butyrate were more effective than higher concentrations. This was reflected in an enhanced decrease in c-Myc and Bax protein in response to mitomycin C with low concentrations of butyrate. Our data indicate that the differentiation stimulus of butyrate, in association with p21(waf1/cip1) induction, and apoptosis, may explain antineoplastic effects of butyrate. Co-carcinogenic features of butyrate may result from inhibition of p53 mediated DNA damage response. PMID- 9333014 TI - Gamma-heregulin: a novel heregulin isoform that is an autocrine growth factor for the human breast cancer cell line, MDA-MB-175. AB - A novel neuregulin isoform, termed gamma-HRG, was cloned and characterized from the human breast cancer cell line, MDA-MB-175. As observed with other neuregulins, gamma-HRG, is a product of alternative mRNA splicing of the neuregulin gene. Gamma-HRG contains the EGF-like and immunoglobulin-like domains that are commonly found in other family members, but lacks a transmembrane and cytoplasmic region. The new isoform possesses a unique N-terminal region that includes a hydrophobic domain that may function as a secretion signal. A purified recombinant version of gamma-HRG competes for binding to soluble ErbB3- and ErbB4 IgG fusion proteins with affinities similar to those observed for rHRGbeta1(177 244). Gamma-HRG has a wide distribution in mesenchymal or neuronal tissues but in contrast to other neuregulins, it is not present in breast, lung, liver and small intestine. Expression of gamma-HRG with its cognate receptors, ErbB3 and ErbB2 suggested that the growth of the MDA-MB-175 cell line might be a result of the autocrine stimulation of a growth factor signaling pathway. Treatment of MDA-MB 175 cells with an anti-ErbB2 monoclonal antibody that interferes with the ligand dependent formation of ErbB2-ErbB3 heterodimer complexes shows a strong growth inhibitory effect on this cell line. Moreover, incubation with a receptor-IgG fusion protein that neutralizes secreted gamma-HRG, also inhibits cell growth. These data suggest that the secretion of gamma-HRG by MDA-MB-175 cells leads to the formation of a constitutively active receptor complex and stimulates the growth of these cells in an autocrine manner. PMID- 9333016 TI - Counter-selection for over-expressed human CD44s in primary tumors versus lung metastases in a mouse fibrosarcoma model. AB - Human CD44 standard isoform cDNA (hCD44s) was transfected into sis-transformed Balb/c 3T3 cells and into ras-revertant IIIA4 cells (both tumorigenic but nonmetastatic). Transfectants were injected subcutaneously into athymic nude mice to elucidate the functional role of hCD44s over-expression in progression and metastasis. The transfectants (but not parental cells) were capable of lung micrometastasis and of binding exogenously-added hyaluronan. hCD44s protein expression was conserved in lung micrometastases suggesting that it may have been necessary for their formation. In contrast, no hCD44s protein was detected in large subcutaneous (s.c.) tumors but normal levels of murine CD44 were detected. A second round of tumor development, using these two tumor cell classes, demonstrated that hCD44s-nonexpressing s.c. tumor cells re-expressed it in lung micrometastases. Conversely, hCD44s-expressing lung micrometastatic cells, when injected into a second group of mice, downregulated hCD44s expression in order to grow sizable s.c. tumors. S.c. tumor cells still contained the hCD44s gene but its expression was inhibited by epigenetic mechanisms, one of which was shown to be methylation of the hCD44s gene. These studies demonstrate (a) opposing selective pressures on CD44s over-expression for s.c. tumor growth and for metastatic spread to the lung and (b) further credence for the significance of CD44 for metastatic spread of fibrosarcomas. Therefore, CD44s may be a critical component of the metastatic phenotype induced by specific oncogenes. PMID- 9333017 TI - Fibroblast growth factor-5 stimulates mitogenic signaling and is overexpressed in human pancreatic cancer: evidence for autocrine and paracrine actions. AB - Fibroblast growth factor (FGF)-1 and -2 are overexpressed in human pancreatic cancer. In this study the role of FGF-5 in human pancreatic cancer was investigated, as FGF-5 has a classical signal sequence for secretion not found in FGF-1 or -2. Northern blot analysis with a 306 bp FGF-5 cDNA revealed the presence of 4.0 kb and 1.6 kb FGF-5 mRNA transcripts in both normal and cancerous pancreatic tissues. Densitometric analysis indicated that 4.0 kb and 1.6 kb FGF-5 mRNA transcripts levels were increased 2.4- and 2.7-fold in the cancers by comparison with normal tissues, respectively (P < 0.002, P < 0.0001). Immunohistochemistry and in situ hybridization demonstrated that FGF-5 localized in the cancer cells, stromal fibroblast and inflitrating macrophages. FGF-5 mRNA was also detected in COLO-357 human pancreatic cancer cells. Furthermore, secreted FGF-5 protein was present in conditioned medium of COLO-357 cells. Exogeneous FGF-5 (0.37 nM) increased the growth of COLO-357 cells by 48% (P < 0.0001) and increased mitogen-activated protein kinase activity. COLO-357 cells expressed the IIIc isoform of the type I FGF receptor, the preferred FGF receptor for FGF-5. These observations suggest that FGF-5 may participate in autocrine and paracrine pathways promoting pancreatic cancer cell growth in vivo. PMID- 9333018 TI - Activation of multiple growth regulatory genes following inducible expression of IRF-1 or IRF/RelA fusion proteins. AB - Interferon Regulatory Factor (IRF)-1 has been characterized as an important growth regulatory and immunomodulatory transcription factor. To further characterize the potential targets of IRF-1 antiproliferative activity, IRF-1 was expressed under the control of the tetracycline-inducible system in murine NIH3T3 cells. Due to their ability to mimic IRF-1 transactivator function, the regulatory potential of IRF-1/RelA and IRF-2/RelA fusion proteins consisting of the DNA binding domains of IRF-1 or IRF-2 fused to the transactivation domain of NF-kappaB RelA(p65) was also examined. Cells inducibly expressing IRF-1 or IRF/RelA in response to doxycycline treatment displayed significantly reduced growth rates compared to control cells, and inhibition of cell growth correlated directly with the level of transgene expression. Interestingly, IRF-1 and IRF/RelA expression also induced a low level of apoptosis, as detected by microscopic analyses. Furthermore, expression of the interferon inducible, double stranded RNA dependent kinase PKR was increased following IRF-1 or IRF/RelA induction. Most strikingly, induction of IRF-1 and IRF/RelA expression resulted in a significant increase in STAT1 (p91) protein and increased ISGF3 DNA binding activity, suggesting that IRF-1 tumor suppressor activity may involve a novel mechanism which activates the JAK-STAT pathway through STAT1. WAF1 levels were also constitutively elevated in IRF-1 and IRF-1/RelA cells. These studies demonstrate that inducible expression of the transactivation function of IRF-1 or IRF/RelA mediates tumor suppressor activity by inducing cell growth arrest, apoptosis and the differential activation of growth regulatory genes such as PKR, STAT1 and WAF1. PMID- 9333019 TI - Enhancement of EGF- and PMA-mediated MAP kinase activation in cells expressing the human papillomavirus type 16 E5 protein. AB - In this report we demonstrate that cells expressing the human papillomavirus type 16 E5 open reading frame (HPV16-E5) show a greatly enhanced transcription of the immediate early genes after EGF or PMA treatment compared to control cells. This enhancement is due to amplification of the signal transduction pathways in response to growth factors or phorbol esters. Upon short-time EGF treatment of the E5-expressing cells we observed an increase in the activation of EGF receptors, resulting in a stronger activation of MAP kinases ERK1/2 compared to control-transfected cells. We also observed that in E5-expressing cells, treatment with PMA results in an increase in membrane-associated PKC activity, and a superactivation of the ERK1/2 MAP kinases. This superactivation is PKC dependent, since pretreatment of the cells with the PKC inhibitor Ro 31-8220 inhibits MAP kinase activation and early gene transcription almost completely. Furthermore, treatment with genistein strongly reduces the PMA-mediated superactivation of ERK1/2 kinases, demonstrating a PKC-mediated, tyrosine kinase dependent pathway in the superinduction of MAP kinase activation. Thus, HPV16-E5 effects superactivation of MAP kinases over at least two different pathways, a PKC-mediated, and another, receptor tyrosine-kinase mediated, PKC-independent one. PMID- 9333021 TI - Continued expression of a tissue specific activated oncogene in the early steps of radiation-induced human thyroid carcinogenesis. AB - Ionizing radiation is a well-known risk factor of cancer development, but the mechanism of radiation induced carcinogenesis is not clear. Chromosomal rearrangements induced by radiation most likely are one of the principal genetic alterations resulting in malignant transformation. The chimeric BCR-ABL associated with chronic myelogenous leukemia (CML) and H4-RET oncogenes associated with thyroid papillary carcinoma are the result of a translocation and inversion, respectively. In vitro studies showed these genes were induced by high doses of X-irradiation in cell lines. Studies also show that therapeutic external X-ray doses as high as 60 Gy for treatment of various childhood cancers including Hodgkin's disease significantly increase the risk of thyroid cancer. Therefore, we examined the induction and persistence of these chimeric genes in human thyroid tissues transplanted in scid mice after 50 Gy exposure as a function of time for 2 months to elucidate the early events of thyroid carcinogenesis. The H4 RET genes were detected on day 2 and throughout the 2 month period. On the other hand, BCR-ABL genes were detected on day 2 and were undetectable subsequently. These results suggest that ionizing radiation causes various oncogene activations, but cells with only specific gene alteration uniquely associated with thyroid carcinogenesis are selectively retained demonstrating one of the early events in the beginnings of radiation carcinogenesis in human thyroid tissues. PMID- 9333020 TI - p16MTS1/CDK4I mutations and concomitant loss of heterozygosity at 9p21-23 are frequent events in squamous cell carcinoma of the larynx. AB - We have examined the presence of p16MTS1/CDK4I gene deletions, mutations and methylation status, and 9p21-23 deletions in a series of 46 squamous cell carcinomas of the larynx and paired normal mucosa previously characterized for cyclin D1 gene amplification and overexpression. pRb expression was also examined by immunohistochemistry. p16MTS1/CDK4I mutations were found in 10/46 (22%) carcinomas and hypermethylation in 2/31 (7%). Loss of heterozygosity at 9p21-23 was found in 24 out of 42 (57%) carcinomas examined. All p16MTS1/CDK4I mutated cases and the two hypermethylated carcinomas showed 9p21-23 loss of heterozygosity. The loss of heterozygosity correlated with advanced local invasion (P=0.0045), lymph node metastases (P=0.0326), stage IV of the tumors (P=0.0058), and existence of cyclin D1 amplification/overexpression (P < 0.03). Only one out of 37 carcinomas was negative for pRb expression. No alterations in p16 gene or 9p21-23 loss of heterozygosity were detected in this case. These findings indicate that p16MTS1/CDK4I is frequently inactivated by gene mutation, hypermethylation, and allelic deletions in a significant subset of squamous cell carcinomas of larynx. Since 9p21-23 loss of heterozygosity was more frequently detected than p16MTS1/CDK4I mutations, and mutated carcinomas invariably had loss of heterozygosity, allelic losses probably precede the p16MTS1/CDK4I mutations. Their association with cyclin D1 deregulation in advanced carcinomas could indicate a possible cooperative effect in the progression of these neoplasms. PMID- 9333022 TI - Resistance to radiation-induced apoptosis in Bcl-2-expressing cells is reversed by depleting cellular thiols. AB - The mechanism by which Bcl-2 oncogene expression inhibits radiation-induced apoptosis has been investigated in two mouse lymphoma cell lines: line LY-as is radiation sensitive, displays substantial radiaton-induced apoptosis, and expresses low levels of Bcl-2; line LY-ar is radiation-resistant, displays a low apoptosis propensity, and expresses 30-fold higher amount of Bcl-2 protein than does the sensitive line. We observed that upon incubation in cystine/methionine free (C/M-) medium, radiation-induced apoptosis in the LY-ar cells was restored to levels comparable to that seen in the LY-as cells. lntracellular glutathione (GSH) concentrations in LY-ar cells incubated in C/M- medium plummeted to 50% of control values within 2 h. LY-ar cells treated with diethyl maleate (DEM) or diamide, agents that deplete cellular thiols, had increased susceptibility to radiation-induced apoptosis in a manner similar to C/M- medium. These results are consistent with the general idea that Bcl-2 expression blocks apoptosis through an antioxidant pathway that involves cellular thiols. That Bcl-2-expressing tumor cells can be sensitized by exogeneous agents that modify cellular thiols offers strategies for overcoming such resistance. PMID- 9333023 TI - Pim-1 kinase stimulates c-Myc-mediated death signaling upstream of caspase-3 (CPP32)-like protease activation. AB - Pim-1 oncoprotein is a serine/threonine kinase that can closely cooperate with c Myc in lymphomagenesis, as does Bcl-2. Although the molecular mechanism of this cooperative transformation remains unknown, it is speculated that, similar to Bcl 2, Pim-1 contributes to transformation by inhibiting apoptosis. In this study, therefore, we examined the effect of Pim-1 expression on c-Myc-mediated apoptosis of Rat-1 fibroblasts triggered by serum deprivation. Our results showed that, rather than inhibiting apoptosis, Pim-1 expression stimulated c-Myc-mediated apoptosis in Rat-1 fibroblasts. Pim-1 stimulated c-Myc-mediated apoptosis through an enhancement of the c-Myc-mediated activation of caspase-3 (CPP32)-like proteases, since the suppression of this activity by a specific caspase inhibitor abolished the apoptosis stimulation by Pim-1. A kinase-defective Pim-1 mutant failed to stimulate c-Myc-mediated apoptosis, and Pim-1 expression alone in the absence of c-Myc overexpression did not induce apoptosis of serum-deprived Rat-1 cells, indicating that the kinase activity of Pim-1 and the activated c-Myc signaling pathway were required for apoptosis stimulation by Pim-1. Together, these results suggest that Pim-1 oncoprotein stimulates as a serine/threonine kinase the death signaling elicited by c-Myc at a step upstream of caspase-3-like protease activation in Rat-1 fibroblasts. Our results also suggest that Pim-1 kinase might function cooperatively with c-Myc through the phosphorylation of a factor(s) which regulates the common signaling pathway involved in c-Myc-mediated apoptosis and transformation. PMID- 9333024 TI - High frequency of simultaneous loss of p16 and p16beta gene expression in squamous cell carcinoma of the esophagus but not in adenocarcinoma of the esophagus or stomach. AB - Quantitative reverse transcription PCR (RT-PCR) was used to measure gene expressions (relative mRNA levels) of p16 and the alternate transcript pl6beta in esophageal and gastric tumors. p16 gene expression was undetectable in 13 of 25 esophageal squamous cell carcinomas. In 11 of these tumors, pl6beta was simultaneously missing whereas two of the pl6-deficient tumors still expressed p16beta. Among 34 esophageal adenocarcinomas and 11 gastric adenocarcinomas, only one tumor lacked p16 expression and all tumors expressed p16beta. p16 sequences were not detectable by PCR in genomic DNA from tumors lacking both p16 and p16beta mRNA, suggesting that the simultaneous loss of both gene expressions resulted from homozygous genomic deletion of the p16 gene. However, DNA from tumors that lacked p16 mRNA but expressed pl6beta did contain the p16 gene, consistent with loss of p16 expression in these tumors by transcriptional suppression. No point mutations in p16 cDNA were detected among 12 that were sequenced, but one p16 cDNA from a squamous cell carcinoma had a 19-base deletion, possibly indicating a splice-site mutation. Among those tumors that expressed p16 mRNA, the gene expression values of both p16 and pl6beta varied over a wide range. In some cases, p16 expression was detectable but low, suggesting that down-regulation of p16 expression may be used in some cases to achieve the funtional equivalent of gene deletion or transcriptional silencing. These results demonstrate that p16 expression patterns differ based on tumor histology and origin. Homozygous deletion of p16 appears to be common in esophageal squamous cell carcinomas but in adenocarcinomas, both gene deletion and transcriptional silencing of p16 were infrequent. PMID- 9333025 TI - Modulation of ETS-1 transcriptional activity by huUBC9, a ubiquitin-conjugating enzyme. AB - Ets transcription factors have Ets DNA binding domains, that have a winged helix turn-helix structure. ETS-1, the founding member of the family, is regulated by the Ras and Ca2+ signaling pathways and is implicated in various physiological processes leading to cell growth, differentiation and apoptosis. We have identified ETS-1 interacting factors with a yeast two-hybrid screen. The majority of the positive clones turned out to encode the human homologue of the yeast ubiquitin-conjugating enzymes UBC9 and Hus5. In two different yeast assays, ETS-1 interacted with huUBC9. In an in vitro GST 'pull-down' assay, ETS-1 and several other Ets family members complexed with huUBC9. Interestingly, in mammalian cells, coexpression of huUBC9 resulted in a substantial increase in the transcriptional activity of ETS-1. Coexpressed huUBC9 did not affect the ETS-1 protein level, and moreover, a point mutation at Cys93, an amino acid known to be essential for ubiquitination, did not abolish the stimulation of the ETS-1 transcriptional activity. Our results indicate that the modulation of ETS-1 activity by huUBC9 results from processes other than ubiquitination and ETS-1 stabilization. PMID- 9333026 TI - Characterisation and chromosome mapping of the human non receptor tyrosine kinase gene, brk. AB - The brk gene encodes a non-receptor protein tyrosine kinase that consists of single SH3, SH2 and catalytic domains. Although BRK shows strongest sequence similarity to members of the SRC family of PTKs, there are several key structural and regulatory differences that place it on its own amongst non-receptor PTKs. In this study we have isolated genomic DNA clones corresponding to the human brk locus and used these to determine the intron-exon structure of the brk gene. The genomic structure of brk consists of 8 exons, whose boundaries are distinct from other non-receptor PTK family members, again indicating a structural and functional divergence. Alternate splicing of the primary brk transcript generates a distinct mRNA which encodes a truncated protein consisting of an SH3 domain and a novel C-terminal proline rich sequence. Using an antiserum raised to the SH3 domain, we have demonstrated that the product of this alternate brk transcript is expressed in the human breast tumour cell line T-47D. We have previously reported that expression of a tumour derived brk cDNA in mouse embryonic fibroblasts and human mammary epithelial cells supports anchorage independent growth, and in the latter potentiates the mitogenic response to epidermal growth factor. The protein encoded by the genomic sequence derived from normal human tissue is identical to that encoded by the tumour derived cDNA, and therefore the altered growth regulation is not associated with mutations within brk. In addition, we have identified a 5' genomic region that has promoter activity. The brk gene has been assigned to chromosome 20q 13.3 [corrected] using fluorescence in situ hybridisation (FISH). PMID- 9333029 TI - Pharmacokinetic and pharmacodynamic models of the antistaphylococcal effects of meropenem and cloxacillin in vitro and in experimental infection. AB - The efficacies of meropenem (MPM) and cloxacillin (CLC) against two Staphylococcus aureus strains were established in vitro. A pharmacodynamic model equation, based on the concept that the killing rate depends on concentration and time, was fitted to the numbers of CFU. The parameters of the equation are maximum killing rate, time point of maximum killing, and 50% effective concentration (EC50). The EC50s for the two strains were 0.047 and 0.040 mg/liter, respectively, for MPM and 0.105 and 0.121 mg/liter, respectively, for CLC. Calculated values of the parameters were used to predict the numbers of CFU at exponentially decreasing concentrations in vitro as well as in an experimental infection model. The prediction for in vitro conditions gave a satisfactory fit (R2, between 0.862 and 0.894). In vivo the numbers were predicted with the assumption that killing rate in vivo is proportional to that in vitro (R2, between 0.731 and 0.973). The proportionality factor ranged between 0.23 and 0.42; this variation was due mainly to covariation with growth rates in control animals, without other significant differences between antibiotics or strains. PMID- 9333028 TI - In vivo antiviral activity and pharmacokinetics of (-)-cis-5-fluoro-1-[2 (hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine in woodchuck hepatitis virus infected woodchucks. AB - The (-) enantiomer of cis-5-fluoro-1l-[2-(hydroxymethyl)-1,3-oxathiolan-5 yl]cytosine [(-)-FTC)], a substituted oxathiolane compound with anti-hepatitis B virus activity in vitro, was assessed for its efficacy in woodchucks with naturally acquired woodchuck hepatitis virus (WHV) infection. Pharmacokinetics and in vitro anabolism were also determined. (-)-FTC was anabolized to the 5' triphosphate in a dose-related fashion, reaching a maximum concentration at about 24 h in cultured woodchuck hepatocytes. Following administration of a dose of 10 mg/kg of body weight intraperitoneally (i.p.), the clearance of (-)-FTC from plasma was monoexponential, the terminal half-life was 3.76 +/- 1.4 h, and the systemic clearance was 0.12 +/- 0.06 liters/h/kg. The antiviral efficacy of (-) FTC in the woodchuck model was assessed by quantitation of serum WHV DNA levels and by WHV particle-associated DNA polymerase activity at two dosages, 30 and 20 mg/kg given i.p. twice daily (b.i.d.), respectively. The level of WHV DNA in serum was reduced 20- to 150-fold (average, 56-fold) in the 30-mg/kg-b.i.d. treatment group and 6- to 49-fold (average, 27-fold) in the 20-mg/kg-b.i.d. treatment group. Viral DNA polymerase levels diminished accordingly. One week after treatment was discontinued, WHV levels returned to pretreatment levels in both studies. These animals were biopsied before and following treatment with 30 mg of (-)-FTC per kg. Their livers were characterized by a mild increase in cytoplasmic lipid levels, but this change was not associated with altered liver enzyme levels. Serum chemistry and hematology results were within the normal ranges for all treated animals. We conclude that (-)-FTC is a potent antihepadnaviral agent and that it has no detectable toxic effects in woodchucks when given for up to 25 days. Further development of (-)-FTC as an anti-hepatitis B virus therapy for patients is warranted. PMID- 9333027 TI - Regulation of chromosomally mediated multiple antibiotic resistance: the mar regulon. PMID- 9333030 TI - Comparison of the efficacies of various formulations of amphotericin B against murine visceral leishmaniasis. AB - The antileishmanial efficacies of four proprietary amphotericin B (AmB) formulations (Fungizone, AmBisome, Abelcet, and Amphocil) and an experimental nonionic surfactant vesicle (NIV) formulation were compared in a murine model of acute visceral leishmaniasis. By a multiple-dosing regimen, groups of Leishmania donovani-infected BALB/c mice were treated (2.5 mg of AmB per kg of body weight) on days 7 to 11 postinfection with one of the AmB formulations, and parasite burdens were determined on day 18 postinfection. All of the formulations caused significant suppression parasite burdens in spleens (P < 0.01 to 0.0005) and livers (P < 0.0005) compared with those in the spleens and livers of the controls. In addition, a significant suppression of parasite burdens in bone marrow (P < 0.0005) compared to the burdens in the bone marrow of the controls was obtained for all the formulations except Abelcet, which was inactive at this site. On the basis of their overall efficacies (activity against liver, spleen, and bone marrow parasites), the formulations could be ranked as follows: Amphocil = AmBisome > AmB-NIV > Abelcet >> Fungizone. On the basis of spectrophotometric measurements, AmB was shown to exist in a predominantly aggregated state in all of the formulations. Although incubation in 50% serum altered the degree of aggregation, the AmB remained predominantly aggregated, indicating that the AMB lipid complex in all of the formulations was physically stable. The results of the study showed that antiparasitic efficacy is associated positively with the degree of AmB aggregation in the presence of serum. PMID- 9333031 TI - Evaluation of the INNO-LiPA Rif. TB assay, a reverse hybridization assay for the simultaneous detection of Mycobacterium tuberculosis complex and its resistance to rifampin. AB - Mycobacterium tuberculosis resistance to rifampin results from nucleotide changes in the gene encoding the beta-subunit of the RNA polymerase (rpoB). We developed a reverse hybridization-based line probe assay (LiPA; the INNO-LiPA Rif. TB) carrying one oligonucleotide probe for the detection of M. tuberculosis complex strains and nine probes designed to detect nucleotide changes in the relevant part of rpoB. This assay was evaluated with 107 M. tuberculosis isolates with known rpoB sequences, 52 non-M. tuberculosis complex strains, and 61 and 203 clinical isolates found to be sensitive and resistant, respectively, by in vitro testing. The results indicated that (i) the M. tuberculosis complex probe was 100% specific, (ii) when compared to the results of nucleotide sequencing, no discrepancies with the results of INNO-LiPA Rif. TB were observed, (iii) all strains sensitive by in vitro susceptibility testing were correctly identified, and (iv) among the strains resistant by in vitro susceptibility testing, only 4 (2%) yielded conflicting results. The INNO-LiPA Rif. TB is therefore a reliable and widely applicable assay and a valuable tool for routine diagnostic use, given its simplicity and rapid performance. PMID- 9333032 TI - Cellular accumulation of the new ketolide RU 64004 by human neutrophils: comparison with that of azithromycin and roxithromycin. AB - We analyzed the uptake of RU 64004 by human neutrophils (polymorphonuclear leukocytes [PMNs]) relative to those of azithromycin and roxithromycin. RU 64004 was strongly and rapidly accumulated by PMNs, with a cellular concentration/extracellular concentration ratio (C/E) of greater than 200 in the first 5 min, and this was followed by a plateau at 120 to 180 min, with a C/E of 461 +/- 14.8 (10 experiments) at 180 min. RU 64004 uptake was moderately sensitive to external pH, and activation energy was also moderate (63 +/- 3.8 kJ/mol). RU 64004 was mainly located in PMN granules (about 70%) and egressed slowly from loaded cells, owing to avid reuptake. The possibility that PMN uptake of RU 64004 and other macrolides occurs through a carrier-mediated system was suggested by three key results. First, there existed a strong interindividual variability in uptake kinetics, suggesting variability in the numbers or activity of a transport protein. Second, macrolide uptake displayed saturation kinetics characteristic of that of a carrier-mediated transport system: RU 64004 had the highest Vmax value (3,846 ng/2.5 x 10(6) PMNs/5 min) and the lowest Km value (about 28 microM), indicating a high affinity for the transporter. Third, as observed previously with other erythromycin A derivatives, Ni2+ (a blocker of the Na+/Ca2+ exchanger which mediates Ca2+ influx in resting neutrophils) impaired RU 64004 uptake by PMNs, with a 50% inhibitory concentration of about 3.5 mM. In addition, we found that an active process is also involved in macrolide efflux, because verapamil significantly potentiated the release of all three macrolides tested. This effect of verapamil does not seem to be related to an inhibition of Ca2+ influx, because neither EGTA [ethylene glycol-bis (beta-aminoethyl ether) N,N',N'-tetraacetic acid] nor Ni2+ modified macrolide efflux. The nature and characteristics of the entry- and efflux-mediating carrier systems are under investigation. PMID- 9333033 TI - In vivo efficacies of 5'-methylthioadenosine analogs as trypanocides. AB - 5'-Deoxy-5'-(methylthio)adenosine (MTA), a key by-product of polyamine biosynthesis, is cleaved by MTA phosphorylase and is salvaged as adenine and, through conversion of the ribose moiety, methionine. An analog of MTA, 5'-deoxy 5'-(hydroxyethylthio)adenosine (HETA), is a substrate for trypanosome MTA phosphorylase and is active in vitro and in vivo against Trypanosoma brucei brucei, an agent of bovine trypanosomiasis. In this study, HETA and three O acylated HETA derivatives were examined for their activities against model infections of T. b. brucei and Trypanosoma brucei rhodesiense, the agent of East African sleeping sickness. HETA was curative (>60%) for infections caused by 5 of 11 clinical isolates of T. b. rhodesiense when it was given to mice at 200 mg/kg of body weight for 7 days as a continuous infusion in osmotic pumps. HETA at 150 to 200 mg/kg also extended the life spans of the mice infected with four additional isolates two- to fivefold. Di- and tri-O-acetylated derivatives of HETA also proved curative for the infections, while a tri-O-propionyl derivative, although also curative, was not as effective. This study indicates that substrate analogs of MTA should be given important consideration for development as novel chemotherapies against African trypanosomiasis. PMID- 9333034 TI - The signal molecule for beta-lactamase induction in Enterobacter cloacae is the anhydromuramyl-pentapeptide. AB - Beta-lactamase induction in Enterobacter cloacae, which is linked to peptidoglycan recycling, was investigated by high-performance liquid chromatographic analysis of cell wall fragments in genetically defined cells of Escherichia coli. After treatment of cells with beta-lactams, we detected an increase in a D-tripeptide (disaccharide-tripeptide, N-acetylglucosaminyl-1,6 anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid), aD tetrapeptide (disaccharide-tetrapeptide, N-acetylglucosaminyl-1,6-anhydro-N acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid-D-alanine), and aD pentapeptide (disaccharide-pentapeptide, N-acetylglucosaminyl-1,6-anhydro-N acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid-D-alanyl-D alanine)levels in the periplasms of bacterial cells. Furthermore, only the accumulation of aD-pentapeptide correlates with the beta-lactamase-inducing capacity of the beta-lactam antibiotic. The transmembrane protein AmpG transports all three aD-peptides into the cytoplasm, where they are degraded into the corresponding monosaccharide peptides. In the absence of AmpD the constitutive overproduction of beta-lactamase is accompanied by an accumulation of aM tripeptide (monosaccharide-tripeptide, anhydro-N-acetylmuramyl-L-alanyl-D glutamyl-meso-diaminopimelic acid) and aM-pentapeptide (L1,6-anhydro-N acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanyl-D-alanine), but not aM-tetrapeptide (anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-meso diaminopimelic acid-D-alanine), in the cytoplasm. Only the amount of aM pentapeptide is increased upon treatment with imipenem. These findings indicate that aD-pentapeptide is the main periplasmic muropeptide, which is converted into the cytoplasmic signal molecule for beta-lactamase induction, the aM pentapeptide. PMID- 9333035 TI - Mechanism of sulfonamide resistance in clinical isolates of Streptococcus pneumoniae. AB - The genetic basis of sulfonamide resistance in six clinical isolates of Streptococcus pneumoniae was demonstrated to be 3- or 6-bp duplications within sulA, the chromosomal gene encoding dihydropteroate synthase. The duplications all result in repetition of one or two amino acids in the region from Arg58 to Tyr63, close to but distinct from the sul-d mutation, a duplication previously reported in a resistant laboratory strain (P. Lopez, M. Espinosa, B. Greenberg, and S. A. Lacks, J. Bacteriol. 169:4320-4326, 1987). Six sulfonamide-susceptible clinical isolates lacked such duplications. The role of the duplications in conferring sulfonamide resistance was confirmed by transforming 319- or 322-bp PCR fragments into the chromosome of a susceptible recipient. Two members of a clone of serotype 9V, one susceptible and one resistant to sulfonamide, which are highly related by other criteria, were shown to have sulA sequences that differ in 7.2% of nucleotides in addition to the duplication responsible for resistance. It is postulated that horizontal gene exchange has been involved in the acquisition (or loss) of resistance within this clone. However, five of the six resistant isolates have distinct duplications and other sequence polymorphisms, suggesting that resistance has arisen independently on many occasions. PMID- 9333036 TI - The oxazolidinone eperezolid binds to the 50S ribosomal subunit and competes with binding of chloramphenicol and lincomycin. AB - The oxazolidinones are a novel class of antibiotics that act by inhibiting protein synthesis. It as been reported that the drug exerts its primary activity on the initiation phase of translation. In order to study the possibility of direct interaction between the drug and the ribosome, we have developed a binding assay using 14C-labelled eperezolid (PNU-100592; formerly U-100592). Eperezolid binds specifically to the 50S ribosomal subunit of Escherichia coli. The specific binding of eperezolid is dose dependent and is proportional to the ribosome concentrations. Scatchard analysis of the binding data reveals that the dissociation constant (Kd) is about 20 microM. The binding of eperezolid to the ribosome is competitively inhibited by chloramphenicol and lincomycin. However, unlike chloramphenicol and lincomycin, eperezolid does not inhibit the puromycin reaction, indicating that the oxazolidinones have no effect on peptidyl transferase. In addition, whereas lincomycin and, to some extent, chloramphenicol inhibit translation termination, eperezolid has no effect. Therefore, we conclude that the oxazolidinones inhibit protein synthesis by binding to the 50S ribosomal subunit at a site close to the site(s) to which chloramphenicol and lincomycin bind but that the oxazolidinones are mechanistically distinct from these two antibiotics. PMID- 9333037 TI - Mechanism of action of oxazolidinones: effects of linezolid and eperezolid on translation reactions. AB - The oxazolidinones are a new class of synthetic antibiotics with good activity against gram-positive pathogenic bacteria. Experiments with a susceptible Escherichia coli strain, UC6782, demonstrated that in vivo protein synthesis was inhibited by both eperezolid (formerly U-100592) and linezolid (formerly U 100766). Both linezolid and eperezolid were potent inhibitors of cell-free transcription-translation in E. coli, exhibiting 50% inhibitory concentrations (IC50s) of 1.8 and 2.5 microM, respectively. The ability to demonstrate inhibition of in vitro translation directed by phage MS2 RNA was greatly dependent upon the amount of RNA added to the assay. For eperezolid, 128 microg of RNA per ml produced an IC50 of 50 microM whereas a concentration of 32 microg/ml yielded an IC50 of 20 microM. Investigating lower RNA template concentrations in linezolid inhibition experiments revealed that 32 and 8 microg of MS2 phage RNA per ml produced IC50s of 24 and 15 microM, respectively. This phenomenon was shared by the translation initiation inhibitor kasugamycin but not by streptomycin. Neither oxazolidinone inhibited the formation of N formylmethionyl-tRNA, elongation, or termination reactions of bacterial translation. The oxazolidinones appear to inhibit bacterial translation at the initiation phase of protein synthesis. PMID- 9333038 TI - The ketolide antibiotics HMR 3647 and HMR 3004 are active against Toxoplasma gondii in vitro and in murine models of infection. AB - Ketolides are a new class of macrolide antibiotics that have been shown to be active against a variety of bacteria including macrolide-resistant bacteria and mycobacteria. We examined two ketolides, HMR 3647 and HMR 3004, for their in vitro and in vivo activities against the protozoan parasite Toxoplasma gondii. In vitro, both ketolides at concentrations as low as 0.05 microg/ml markedly inhibited replication of tachyzoites of the RH strain within human foreskin fibroblasts. HMR 3004 demonstrated some toxicity for host cells after they were exposed to 5 microg of the drug per ml for 72 h. In contrast, HMR 3647 did not show any significant toxicity even at concentrations as high as 25 microg/ml. In vivo, both ketolides provided remarkable protection against death in mice lethally infected intraperitoneally with tachyzoites of the RH strain or orally with tissue cysts of the C56 strain of T. gondii. A dosage of 100 mg of HMR 3647 per kg of body weight per day administered for 10 days protected 50% of mice infected with tachyzoites. The same dosage of HMR 3004 protected 100% of the mice. In mice infected with cysts, a dosage of 30 mg of HMR 3647 per kg per day protected 100% of the mice, whereas a dosage of 40 mg of HMR 3004 per kg per day protected 75% of the mice. These results demonstrate that HMR 3647 and HMR 3004 possess excellent activities against two different strains of T. gondii and may be useful for the treatment of toxoplasmosis in humans. PMID- 9333039 TI - Phosphorothioate oligonucleotides derived from human immunodeficiency virus type 1 (HIV-1) primer tRNALys3 are strong inhibitors of HIV-1 reverse transcriptase and arrest viral replication in infected cells. AB - Retroviral reverse transcriptase (RT) is involved in the selection of a specific tRNA primer which initiates proviral DNA minus-strand synthesis. Studies of the interactions between human immunodeficiency virus type 1 (HIV-1) RT and primer tRNALys3 have shown that the dihydrouridine (diHU), anticodon, and pseudouridine regions of tRNA are highly protected in the RT-tRNA complex. The CCA 3' end of tRNA is also in close contact with the enzyme during the cDNA initiation step. Using synthetic oligoribonucleotides corresponding to the anticodon and diHU regions, we have previously shown a low but significant inhibition of HIV-1 RT activity. We extend this observation and show that primer tRNA-derived oligodeoxynucleotides (ODNs) carrying a phosphorothioate (PS) modification are strong inhibitors of HIV-1 RT. The affinity of PS-ODNs for the enzyme was monitored by gel mobility shift electrophoresis. Experiments with HIV-1-infected human cells (MT-2 cells) were performed with the latter ODNs. A PS-ODN corresponding to the 3' end of tRNALys3 (acceptor stem [AS]) was able to inhibit HIV-1 replication. No effect of the other modified ODNs was observed in infected cells. The analysis of HIV-1 RNase H activity in a cell-free system strongly suggests that the inhibitory effect of the PS-AS may be mediated via both a sense and an antisense mechanism. PMID- 9333040 TI - Antibacterial activity of RU 64004 (HMR 3004), a novel ketolide derivative active against respiratory pathogens. AB - The antibacterial activity of RU 64004, a new ketolide, was evaluated against more than 600 bacterial strains and was compared with those of various macrolides and pristinamycin. RU 64004 had good activity against multiresistant pneumococci, whether they were erythromycin A resistant or not, including penicillin-resistant strains. RU 64004 inhibited 90% of pneumococci resistant to erythromycin A and penicillin G at 0.6 and 0.15 microg/ml, respectively. Unlike macrolides, RU 64004 did not induce the phenotype of resistance to macrolides-lincosamides streptogramin B. Its good antibacterial activity against multiresistant pneumococci ran in parallel with its well-balanced activity against all bacteria involved in respiratory infections (e.g., Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pyogenes). In contrast to all comparators (14- and 16 membered-ring macrolides and pristinamycin), RU 64004 displayed high therapeutic activity in animals infected with all major strains, irrespective of the phenotypes of the strains. The results suggest that RU 64004 has potential for use in the treatment of infections caused by respiratory pathogens including multiresistant pneumococci. PMID- 9333041 TI - Activities of the human immunodeficiency virus type 1 (HIV-1) protease inhibitor nelfinavir mesylate in combination with reverse transcriptase and protease inhibitors against acute HIV-1 infection in vitro. AB - Nelfinavir mesylate (formerly AG1343) is a potent and selective, nonpeptidic inhibitor of human immunodeficiency virus type 1 (HIV-1) protease that was discovered by protein structure-based design methodologies. We evaluated the antiviral and cytotoxic effects of two-drug combinations of nelfinavir with the clinically approved antiretroviral therapeutics zidovudine (ZDV), lamivudine (3TC), dideoxycytidine (ddC; zalcitabine), stavudine (d4T), didanosine (ddI), indinavir, saquinavir, and ritonavir and a three-drug combination of nelfinavir with ZDV and 3TC against an acute HIV-1 strain RF infection of CEM-SS cells in vitro. Quantitative assessment of drug interaction was evaluated by a universal response surface approach (W. R. Greco, G. Bravo, and J. C. Parsons, Pharm. Rev. 47:331-385, 1995) and by the method of M. N. Prichard and C. Shipman (Antiviral Res. 14:181-206, 1990). Both analytical methods yielded similar results and showed that the two-drug combinations of nelfinavir with the reverse transcriptase inhibitors ZDV, 3TC, ddI, d4T, and ddC and the three-drug combination with ZDV and 3TC resulted in additive to statistically significant synergistic interactions. In a similar manner, the combination of nelfinavir with the three protease inhibitors resulted in additive (ritonavir and saquinavir) to slightly antagonistic (indinavir) interactions. In all combinations, minimal cellular cytotoxicity was observed with any drug alone and in combination. These results suggest that administration of combinations of the appropriate doses of nelfinavir with other currently approved antiretroviral therapeutic agents in vivo may result in enhanced antiviral activity with no associated increase in cellular cytotoxicity. PMID- 9333042 TI - In vitro antibacterial activity of LY333328, a new semisynthetic glycopeptide. AB - LY333328 is a semisynthetic N-alkyl derivative of LY264826, a naturally occurring structural analog of vancomycin. LY333328 was evaluated for its in vitro inhibitory and bactericidal activities in comparison with those of the two currently available glycopeptides (vancomycin and teicoplanin). Glycopeptide susceptible test strains included a total of 311 isolates (most of clinical origin) from the genera Staphylococcus, Enterococcus, Streptococcus, Aerococcus, Gemella, Lactococcus, Listeria, Corynebacterium, and Clostridium. Test strains resistant or intermediate to vancomycin and/or teicoplanin included 56 clinical isolates of Enterococcus (of the VanA, VanB, and VanC phenotypes) and 32 clinical isolates of Staphylococcus (S. haemolyticus, S. epidermidis, and S. aureus), 31 strains of gram-positive genera outside the spectrum of activity of vancomycin (Leuconostoc, Pediococcus, Lactobacillus, and Erysipelothrix), and laboratory derived organisms obtained after exposure of susceptible Staphylococcus isolates to teicoplanin (6 strains) or laboratory-derived organisms with resistance determinants received from VanA enterococci (2 Enterococcus and 25 Listeria transconjugants). LY333328 was highly active against staphylococci, enterococci, and listeriae (whether they were clinical or laboratory-derived strains) resistant to the currently available glycopeptides. In particular, the MICs of LY333328 did not vary substantially between teicoplanin-susceptible and teicoplanin-resistant staphylococci and between vancomycin-susceptible and vancomycin-resistant enterococci. LY333328 demonstrated fairly good inhibitory activity even against most strains of Leuconostoc, Pediococcus, and Erysipelothrix (MIC range, 1 to 8 microg/ml), whereas it proved less active (although much more active than vancomycin or teicoplanin) against Lactobacillus strains. In minimal bactericidal concentration (MBC) and time-kill studies, LY333328 demonstrated excellent bactericidal activity; enterococci, in particular, which were largely tolerant of vancomycin and teicoplanin, were uniformly killed by LY333328, with MBC-to-MIC ratios of 4 to 8 for most vancomycin-susceptible and vancomycin-resistant strains. In attempts to select for resistant clones, no survivors stably growing in the presence of 10 microg of LY333328 per ml were obtained from the Staphylococcus and Enterococcus test strains exposed to the drug. PMID- 9333043 TI - Postantibiotic effect of sanfetrinem compared with those of six other agents against 12 penicillin-susceptible and -resistant pneumococci. AB - The postantibiotic effect (PAE) and postantibiotic sub-MIC effect (PAE-SME) of sanfetrinem were compared to those of penicillin G, amoxicillin, cefpodoxime, ceftriaxone, imipenem, and clarithromycin against four penicillin-susceptible, four intermediately susceptible, and four resistant pneumococci. The MICs of imipenem were the lowest against all of the strains (0.03 to 0.5 microg/ml), followed by those of sanfetrinem (0.016 to 1.0 microg/ml), amoxicillin and ceftriaxone (0.016 to 2.0 microg/ml), and cefpodoxime (0.03 to 8.0 microg/ml). High-level resistance to clarithromycin (MIC, >64.0 microg/ml) was seen in three selected strains. The PAEs of all of the oral beta-lactams tested were similar for all of the strains, ranging from 1 to 6.5 h. The PAEs of ceftriaxone and imipenem ranged from 1 to 8 h, and those of clarithromycin ranged from 1 to 7 h. The mean PAEs of all of the beta-lactams and clarithromycin were 2.8 to 4.3 and 2.5 h, respectively. PAE-SMEs could not be determined for all of the strains due to complete killing, especially at high subinhibitory concentrations. However, the overall pattern with all of the compounds tested was that PAE-SMEs were longer than PAEs. Measurable PAE-SMEs of sanfetrinem at the three subinhibitory concentrations (0.125, 0.25, and 0.5 times the MIC) were 2 to 7, 2 to 7, and 3 to 6 h, while those of amoxicillin and cefpodoxime were 1 to 7.5, 2 to 4, and 4 to 9 and 2 to 7, 4 to 7, and 4 to 6 h, respectively. Measurable PAE-SMEs of ceftriaxone and imipenem were 1 to 6.5, 2 to 9, and 2 to 9 and 1.5 to 6, 2 to 5.8, and 4 to 7.7 h, respectively. Measurable clarithromycin PAE-SMEs were 1 to 5, 1 to 5, and 1 to 6 h at the three concentrations. PMID- 9333045 TI - Pharmacokinetics of 1-(2-fluoro-5-methyl-beta-L-arabinofuranosyl)uracil in woodchucks. AB - 1-(2-Fluoro-5-methyl-beta-L-arabinofuranosyl)uracil (L-FMAU) is a nucleoside analog with potent in vitro activity against hepatitis B virus (HBV) and Epstein Barr virus. The purpose of this study was to characterize the disposition of L FMAU following oral and intravenous administration in the woodchuck animal model. The numerous similarities between woodchuck hepatitis virus and HBV infection justify the use of the woodchuck as an animal model for preclinical studies of anti-HBV agents in vivo. Woodchucks were given 25 mg of L-FMAU per kg of body weight intravenously and orally. Concentrations of L-FMAU in urine and plasma were determined by high-performance liquid chromatography. Following intravenous administration of 25 mg of L-FMAU per kg to woodchucks, total clearance was moderate, averaging 0.23 +/- 0.07 liter/h/kg. Renal clearance and nonrenal clearance averaged 0.13 +/- 0.08 and 0.10 +/- 0.06 liter/h/kg, respectively. The steady-state volume of distribution averaged 0.99 +/- 0.17 liter/kg, indicative of intracellular distribution of the nucleoside. The terminal-phase half-life of L-FMAU following intravenous administration averaged 6.2 +/- 2.0 h, and mean residence time averaged 4.5 +/- 0.8 h. Absorption of L-FMAU after oral administration was incomplete, and bioavailability was approximately 20%. Concentrations of L-FMAU in plasma remained above the in vitro 50% effective concentration of 0.026 microg/ml for HBV (C. K. Chu, T. Ma, K. Shanmuganathan, C. Wang, Y. Xiang, S. B. Pai, G.-Q. Yao, J.-P. Sommadossi, and Y.-C. Cheng, Antimicrob. Agents Chemother. 39:979-981, 1995) for 24 h after both intravenous and oral administration of 25 mg of L-FMAU per kg. PMID- 9333044 TI - Mechanism of suppression of piperacillin resistance in enterobacteria by tazobactam. AB - Resistance to piperacillin in several isolates of Citrobacter freundii and Enterobacter cloacae was investigated and confirmed to occur at a frequency of 10(-7) to 10(-6). Development of resistance to piperacillin was significantly suppressed by tazobactam but not by clavulanic acid. To elucidate the mechanism by which resistance suppression occurs, the effect of piperacillin plus tazobactam on the induction of AmpC beta-lactamase was analyzed by monitoring the beta-galactosidase activity of an inducible ampC-lacZ gene fusion in Escherichia coli. The combination exerted no inhibitory effect on AmpC beta-lactamase induction. Tazobactam also had no effect on the accumulation of a key intermediate in the AmpC beta-lactamase induction pathway, 1,6 anhydromurotripeptide, in an ampD mutant strain of E. coli. However, the addition of tazobactam to liquid cultures of E. cloacae 40001 in the presence of piperacillin at four times the MIC caused a delay in the recovery of the culture to piperacillin-induced stress. At 16 times the MIC, a complete suppression of regrowth occurred. Analysis of culture viability on piperacillin plates showed that the culture recovery was due to growth by moderately resistant mutants preexisting in the cell population, which at 16 times the MIC became susceptible to the combination. Evidence from the kinetics of inhibition of the E. cloacae 40001 AmpC beta-lactamase by clavulanic acid, sulbactam, and tazobactam and from the effects of these drugs on the frequency of resistance to piperacillin suggests that the suppressive effect of tazobactam on the appearance of resistance is primarily mediated by the beta-lactamase inhibitory activity. PMID- 9333046 TI - OXA-18, a class D clavulanic acid-inhibited extended-spectrum beta-lactamase from Pseudomonas aeruginosa. AB - Clinical isolate Pseudomonas aeruginosa Mus showed resistance both to extended spectrum cephalosporins and to aztreonam. We detected a typical double-disk synergy image when ceftazidime or aztreonam was placed next to a clavulanic acid disk on an agar plate. This resistance phenotype suggested the presence of an extended-spectrum beta-lactamase. Isoelectric focusing revealed that this strain produced three beta-lactamases, of pI 5.5, 7.4, and 8.2. A 2.6-kb Sau3A fragment encoding the extended-spectrum beta-lactamase of pI 5.5 was cloned from P. aeruginosa Mus genomic DNA. This enzyme, named OXA-18, had a relative molecular mass of 30.6 kDa. OXA-18 has a broad substrate profile, hydrolyzing amoxicillin, ticarcillin, cephalothin, ceftazidime, cefotaxime, and aztreonam, but not imipenem or cephamycins. Its activity was totally inhibited by clavulanic acid at 2 microg/ml. Hydrolysis constants of OXA-18 (Vmax, Km) confirmed the MIC results. Cloxacillin and oxacillin hydrolysis was noticeable with the partially purified OXA-18. The blaOXA-18 gene encodes a 275-amino-acid protein which has weak identity with all class D beta-lactamases except OXA-9 and OXA-12 (45 and 42% amino acid identity, respectively). OXA-18 is likely to be chromosomally encoded since no plasmid was found in the strain and because attempts to transfer the resistance marker failed. OXA-18 is peculiar since it is a class D beta-lactamase which confers high resistance to extended-spectrum cephalosporins and seems to have unique hydrolytic properties among non-class A enzymes. PMID- 9333047 TI - Effects of food and sucralfate on a single oral dose of 500 milligrams of levofloxacin in healthy subjects. AB - The effects of food and sucralfate on the pharmacokinetics of levofloxacin following the administration of a single 500-mg oral dose were investigated in a randomized, three-way crossover study with young healthy subjects (12 males and 12 females). Levofloxacin was administered under three conditions: fasting, fed (immediately after a standardized high-fat breakfast), and fasting with sucralfate given 2 h following the administration of levofloxacin. The concentrations of levofloxacin in plasma and urine were determined by high pressure liquid chromatography. By noncompartmental methods, the maximum concentration of drug in serum (Cmax), the time to Cmax (Tmax), the area under the concentration-time curve (AUC), half-life (t1/2), clearance (CL/F), renal clearance (CLR), and cumulative amount of levofloxacin in urine (Ae) were estimated. The individual profiles of the drug concentration in plasma showed little difference among the three treatments. The only consistent effect of the coadministration of levofloxacin with a high-fat meal for most subjects was that levofloxacin absorption was delayed and Cmax was slightly reduced (Tmax, 1.0 and 2.0 h for fasting and fed conditions, respectively [P = 0.002]; Cmax, 5.9 +/- 1.3 and 5.1 +/- 0.9 microg/ml [90% confidence interval = 0.79 to 0.94] for fasting and fed conditions, respectively). Sucralfate, which was administered 2 h after the administration of levofloxacin, appeared to have no effect on levofloxacin's disposition compared with that under the fasting condition. Mean values of Cmax and AUC from time zero to infinity were 6.7 +/- 3.2 microg/ml and 47.9 +/- 8.4 microg x h/ml, respectively, following the administration of sucralfate compared to values of 5.9 +/- 1.3 microg/ml and 50.5 +/- 8.1 microg x h/ml, respectively, under fasting conditions. The mean t1/2, CL/F, CLR, and Ae values were similar among all three treatment groups. In conclusion, the absorption of levofloxacin was slightly delayed by food, although the overall bioavailability of levofloxacin following a high-fat meal was not altered. Finally, sucralfate did not alter the disposition of levofloxacin when sucralfate was given 2 h after the administration of the antibacterial agent, thus preventing a potential drug-drug interaction. PMID- 9333049 TI - In vitro and in vivo antibacterial efficacies of CFC-222, a new fluoroquinolone. AB - CFC-222 is a novel fluoroquinolone containing a C-7 bicyclic amine moiety with potent antibacterial activities against gram-positive, gram-negative, and anaerobic organisms. We compared the in vitro and in vivo activities of CFC-222 with those of ciprofloxacin, ofloxacin, and lomefloxacin. CFC-222 was more active than the other fluoroquinolones tested against gram-positive bacteria. CFC-222 was particularly active against Streptococcus pneumoniae (MIC at which 90% of isolates are inhibited [MIC90], 0.2 microg/ml), Staphylococcus aureus (MIC90, 0.2 microg/ml for ciprofloxacin-susceptible strains), and Enterococcus faecalis (MIC90, 0.39 microg/ml). Against Escherichia coli and other members of the family Enterobacteriaceae, CFC-222 was slightly less active than ciprofloxacin (MIC90s for E. coli, 0.1 and 0.025 microg/ml, respectively). The in vitro activity of CFC 222 was not influenced by inoculum size, medium composition, or the presence of horse serum. However, its activity was decreased significantly by a change in the pH of the medium from 7.0 to 6.0, as was the case for the other quinolones tested. The in vivo protective efficacy of CFC-222 by oral administration was greater than those of the other quinolones tested in a mouse model of intraperitoneally inoculated systemic infection caused by S. aureus. CFC-222 exhibited efficacy comparable to that of ciprofloxacin in the same model of infection caused by gram-negative organisms, such as E. coli and Klebsiella pneumoniae. In this infection model, CFC-222 was slightly less active than ciprofloxacin against Pseudomonas aeruginosa. These results suggest that CFC-222 may be a promising therapeutic agent in various bacterial infections. PMID- 9333048 TI - Pharmacokinetic profile of ABELCET (amphotericin B lipid complex injection): combined experience from phase I and phase II studies. AB - Amphotericin B (AmB) has been the most effective systemic antifungal agent, but its use is limited by the dose-limiting toxicity of the conventional micellar dispersion formulation (Fungizone). New formulations with better and improved safety profiles are being developed and include ABELCET (formerly ABLC), but their dispositions have not been well characterized; hence, the reason for their improved profiles remains unclear. This report details the pharmacokinetics of ABELCET examined in various pharmacokinetic and efficacy studies by using whole blood measurements of AmB concentration performed by high-pressure liquid chromatography. The data indicated that the disposition of AmB after administration of ABELCET is different from that after administration of Fungizone, with a faster clearance and a larger volume of distribution. It exhibits complex and nonlinear pharmacokinetics with wide interindividual variability, extensive distribution, and low clearance. The pharmacokinetics were unusual. Clearance and volume of distribution were increased with dose, peak and trough concentrations after multiple dosings increased less than proportionately with dose, steady state appeared to have been attained in 2 to 3 days, despite an estimated half-life of up to 5 days, and there was no evidence of significant accumulation in the blood. The data are internally consistent, even though they were gathered under different conditions and circumstances. The pharmacokinetics of ABELCET suggest that lower concentrations in blood due to higher clearance and greater distribution may be responsible for its improved toxicity profile compared to those of conventional formulations. PMID- 9333050 TI - In vitro synergism between cefotaxime and minocycline against Vibrio vulnificus. AB - We conducted time-kill studies to evaluate the inhibitory activities of either cefotaxime or minocycline alone and the two drugs in combination against a clinical strain of Vibrio vulnificus. The MICs of cefotaxime and minocycline were 0.03 and 0.06 microg/ml, respectively. When approximately 5 x 10(5) CFU of V. vulnificus per ml was incubated with cefotaxime at 0.03 or 0.05 microg/ml, the bacterial growth was inhibited during the initial 2 and 8 h, respectively. Thereafter, V. vulnificus regrew and the level of growth reached that of the control. Within the dose range of less than five times the MIC, the duration of the inhibitory effect of cefotaxime was proportional to its concentration. When minocycline at 0.015, 0.03, 0.045, and 0.06 microg/ml was used to evaluate the inhibitory effect, a similar trend was observed. Either antibiotic at a concentration of five times the MIC or greater prevented the regrowth of V. vulnificus for at least 48 h. When cefotaxime at 0.05 microg/ml and minocycline at 0.045 microg/ml were combined in the same culture, the inhibitory effect against V. vulnificus persisted for more than 48 h, with no regrowth noted. The use of a combination of these two antibiotics resulted in the reduction of growth by 6 orders of magnitude compared to the use of either of the two antibiotics alone, and the number of surviving organisms in the presence of the antibiotics combined was approximately 3 orders of magnitude less than that in the starting inoculum. We conclude that cefotaxime and minocycline acted synergistically in inhibiting V. vulnificus in vitro. PMID- 9333051 TI - Pharmacodynamic effects of antibiotics and acid pump inhibitors on Helicobacter pylori. AB - Pharmacodynamic studies of Helicobacter pylori exposed to amoxicillin, clarithromycin, metronidazole, omeprazole, and lansoprazole were performed with microscopy, viable count determination, and bioluminescence assay of intracellular ATP. The pharmacodynamic parameters determined were change in morphology, change in cell density, postantibiotic effect (PAE), and control related effective regrowth time (CERT). The PAE is delayed regrowth after brief exposure to antibiotics or acid pump inhibitors. CERT was defined as the time required for the bacteria to resume logarithmic growth and return to the pre exposure inoculum in the test culture minus the corresponding time for the control culture. CERT measures the combined effect of initial killing and PAE. There was a good concordance between the bioluminescence assay and viable counts for determining CERT, which makes this parameter useful for pharmacodynamic studies of the effects of antibiotics and acid pump inhibitors on H. pylori. Amoxicillin and metronidazole produced a strong, concentration-dependent initial decrease in CFU per milliliter, but there was a less prominent initial change in intracellular ATP in these cultures. Amoxicillin caused a long PAE when assayed by the bioluminescence assay but no PAE or a negative PAE when assayed by viable count determination. However, amoxicillin showed similar long CERTs with both methods. The pharmacodynamic effects of amoxicillin were concentration dependent up to a maximum response, indicating that concentrations above this level do not increase the antibiotic effect. The PAEs and CERTs of clarithromycin and metronidazole were concentration dependent with no maximum response. With omeprazole and lanzoprazole, there was no PAE or CERT. PMID- 9333052 TI - Studies of the mechanism of human salivary histatin-5 candidacidal activity with histatin-5 variants and azole-sensitive and -resistant Candida species. AB - Histatins are a group of small, cationic, antifungal peptides present in human saliva. A previous molecular modeling analysis suggested structural similarity between the Phe14-His15 and His18-His19 dipeptide sequences in histatin-5 (Hsn-5; a 24-amino-acid polypeptide) and the sequence of miconazole (one of the azole based antifungal therapeutic agents), implying that the mechanisms of killing of Candida albicans by these two molecules may be similar. To further elaborate on this observation, we have produced two variants of Hsn-5 in which Phe14-His15 or His18-His19 dipeptide sequences were replaced by Ala-Ala (F14A/H15A and H18A/H19A) to eliminate the phenyl and imidazole rings of the side chains and assessed their candidacidal activities against C. albicans. In addition, we tested azole-resistant C. albicans and Candida glabrata strains for their susceptibilities to Hsn-5. Analysis of the purified recombinant proteins for their candidacidal activities indicated that both variants were significantly less effective (the molar concentrations required to kill half of the maximum number of cells [ED50s], approximately 67 and approximately 149 microM for F14A/H15A and H18A/H19A, respectively) than the unaltered Hsn-5 (ED50, approximately 8 microM) at killing C. albicans, suggesting that the two dipeptide sequences are important for the candidacidal activity of Hsn-5. Assessment of the candidacidal activity of Hsn-5 with the well-characterized azole-resistant strains of C. albicans and C. glabrata, however, suggested that the mode of action of histatins against Candida is distinct from that of azole-based antifungal agents because Hsn-5 kills both azole-sensitive and azole-resistant strains equally well. PMID- 9333054 TI - Activity of liposomal nystatin against disseminated Aspergillus fumigatus infection in neutropenic mice. AB - The purpose of this study was to examine the activity of liposomal nystatin against a disseminated Aspergillus fumigatus infection in neutropenic mice. Mice were made neutropenic with 5-fluorouracil and were administered the antifungal drug intravenously for 5 consecutive days beginning 24 h following infection. Liposomal nystatin, at doses as low as 2 mg/kg of body weight/day, protected neutropenic mice against Aspergillus-induced death in a statistically significant manner at the 50-day time point compared to either the no-treatment, the saline, or the empty-liposome group. This protection was approximately the same as that for free nystatin, a positive control. Histopathological results showed that liposomal nystatin cleared the lungs, spleen, pancreas, kidney, and liver of Aspergillus and that there was no organ damage at the day 5 time point, which was after only three doses of liposomal nystatin. Based on these results in mice, it is probable that liposomal nystatin will be effective against Aspergillus infection in humans. PMID- 9333053 TI - Molecular biological characterization of an azole-resistant Candida glabrata isolate. AB - Two isolates of Candida glabrata, one susceptible and one resistant to azole antifungals, were previously shown to differ in quantity and activity of the cytochrome P-450 14alpha-lanosterol demethylase which is the target for azole antifungals. The resistant isolate also had a lower intracellular level of fluconazole, but not of ketoconazole or itraconazole, than the susceptible isolate. In the present study a 3.7-fold increase in the copy number of the CYP51 gene, encoding the 14alpha-lanosterol demethylase, was found. The amount of CYP51 mRNA transcript in the resistant isolate was eight times greater than it was in the susceptible isolate. Hybridization experiments on chromosomal blots indicated that this increase in copy number was due to duplication of the entire chromosome containing the CYP51 gene. The phenotypic instability of the resistant isolate was demonstrated genotypically: a gradual loss of the duplicated chromosome was seen in successive subcultures of the isolate in fluconazole-free medium and correlated with reversion to susceptibility. The greater abundance of the amplified chromosome induced pronounced differences in the protein patterns of the susceptible and revertant isolates versus that of the resistant isolate, as demonstrated by two-dimensional gel electrophoresis (2D-GE). Densitometry of the 2D-GE product indicated upregulation of at least 25 proteins and downregulation of at least 76 proteins in the resistant isolate. PMID- 9333055 TI - Antimicrobial susceptibility patterns of thermophilic Campylobacter spp. from humans, pigs, cattle, and broilers in Denmark. AB - The MICs of 16 antimicrobial agents were determined for 202 Campylobacter jejuni isolates, 123 Campylobacter coli isolates, and 6 Campylobacter lari isolates from humans and food animals in Denmark. The C. jejuni isolates originated from humans (75), broilers (95), cattle (29), and pigs (3); the C. coli isolates originated from humans (7), broilers (17), and pigs (99); and the C. lari isolates originated from broilers (5) and cattle (1). All isolates were susceptible to apramycin, neomycin, and gentamicin. Only a few C. jejuni isolates were resistant to one or more antimicrobial agents. Resistance to tetracycline was more common among C. jejuni isolates from humans (11%) than among C. jejuni isolates from animals (0 to 2%). More resistance to streptomycin was found among C. jejuni isolates from cattle (10%) than among those from humans (4%) or broilers (1%). A greater proportion of C. coli than of C. jejuni isolates were resistant to the other antimicrobial agents tested. Isolates were in most cases either coresistant to tylosin, spiramycin, and erythromycin or susceptible to all three antibiotics. More macrolide-resistant isolates were observed among C. coli isolates from swine (79%) than among C. coli isolates from broilers (18%) and humans (14%). Twenty four percent of C. coli isolates from pigs were resistant to enrofloxacin, whereas 29% of C. coli isolates from humans and none from broilers were resistant. More resistance to streptomycin was observed among C. coli isolates from swine (48%) than among C. coli isolates from broilers (6%) or humans (0%). The six C. lari isolates were susceptible to all antimicrobial agents except ampicillin and nalidixic acid. This study showed that antimicrobial resistance was found only at relatively low frequencies among C. jejuni and C. lari isolates. Among C. coli isolates, especially from swine, there was a high level of resistance to macrolides and streptomycin. Furthermore, this study showed differences in the resistance to antimicrobial agents among Campylobacter isolates of different origins. PMID- 9333056 TI - mefE is necessary for the erythromycin-resistant M phenotype in Streptococcus pneumoniae. AB - Recently, it was shown that a significant number of erythromycin-resistant Streptococcus pneumoniae and Streptococcus pyogenes strains contain a determinant that mediates resistance via a putative efflux pump. The gene encoding the erythromycin-resistant determinant was cloned and sequenced from three strains of S. pneumoniae bearing the M phenotype (macrolide resistant but clindamycin and streptogramin B susceptible). The DNA sequences of mefE were nearly identical, with only 2-nucleotide differences between genes from any two strains. When the mefE sequences were compared to the mefA sequence from S. pyogenes, the two genes were found to be closely related (90% identity). Strains of S. pneumoniae were constructed to confirm that mefE is necessary to confer erythromycin resistance and to explore the substrate specificity of the pump; no substrates other than 14 and 15-membered macrolides were identified. PMID- 9333057 TI - Pharmacokinetic profile of levofloxacin following once-daily 500-milligram oral or intravenous doses. AB - The pharmacokinetics of once-daily oral levofloxacin (study A) or intravenous levofloxacin (study B) in 40 healthy male volunteers were investigated in two separate randomized, double-blind, parallel-design, placebo-controlled studies. Levofloxacin at 500 mg or placebo was administered orally or intravenously as a single dose on day 1; daily oral or intravenous dosing resumed on days 4 to 10. In a third study (study C), the comparability of the bioavailabilities of two oral and one intravenous levofloxacin formulations were investigated with 24 healthy male subjects in an open-label, randomized, three-way crossover study. Levofloxacin at 500 mg as a single tablet or an intravenous infusion was administered on day 1; following a 1-week washout period, subjects received the second regimen (i.e., the other oral formulation or the intravenous infusion); the third and final regimen was administered following a 1-week washout period. The concentrations of drug in plasma and urine were measured by validated high pressure liquid chromatography methods. Pharmacokinetic parameters were estimated by noncompartmental methods. In both study A (oral levofloxacin) and study B (intravenous levofloxacin), steady state was attained within 48 h after the start of the multiple dosing on day 4. Levofloxacin pharmacokinetics were linear and predictable for the single and multiple 500-mg, once-daily oral and intravenous dosing regimens, and the values of the pharmacokinetic parameters for the oral and intravenous administrations were similar. Study C indicated that levofloxacin was rapidly and completely absorbed from the oral tablets, with mean times to the maximum concentration of drug in serum of approximately 1.5 h and mean absolute bioavailability of > or =99%. These results support the interchangeability of the oral and intravenous routes of levofloxacin administration. PMID- 9333058 TI - Chlorproguanil-dapsone: effective treatment for uncomplicated falciparum malaria. AB - Pyrimethamine-sulfadoxine, the first choice for uncomplicated falciparum malaria in Africa, exerts strong selection pressure for resistance because of its slow elimination. It is likely that resistance will emerge rapidly, and there is no widely affordable replacement. Chlorproguanil-dapsone is cheap, rapidly eliminated, more potent than pyrimethamine-sulfadoxine, and could be introduced in the near future to delay the onset of antifolate resistance and as "salvage therapy" for pyrimethamine-sulfadoxine failure. A total of 448 children were randomly allocated (double blind) to either a single dose of pyrimethamine sulfadoxine or to one of two chlorproguanil-dapsone regimens: a single dose or three doses at 24-h intervals. Reinfections are clinically indistinguishable from recrudescence and are more likely after treatment with rapidly eliminated drugs; we measured the incidence of parasitemia in 205 initially aparasitemic children to allow comparison with the three treatment groups. The patients and a community surveillance group were followed up for 28 days. At the study end point, 31.2% (95% confidence interval, 24.9-38.0) of the community surveillance group subjects were parasitemic, compared with subjects in the treatment groups, whose rates of parasitemia were 40.8% (32.9-49.0; relative risk [RR], 1.31 [0.99-1.73]) after triple-dose chlorproguanil-dapsone, 19.7% (13.5-27.2; RR, 0.63 [0.43-0.93]) after pyrimethamine-sulfadoxine, and 65.6% (57.5-73.0; RR, 2.10 [1.66-2.65]) after single-dose chlorproguanil-dapsone. Pyrimethamine-sulfadoxine and triple-dose chlorproguanil-dapsone were effective treatments. Pyrimethamine-sulfadoxine provided chemoprophylaxis during follow-up because of its slow elimination. Triple-dose chlorproguanil-dapsone should now be developed in an attempt to reduce the rate of emergence of antifolate resistance in Africa and for affordable salvage therapy in cases of pyrimethamine-sulfadoxine failure. PMID- 9333059 TI - Widespread detection of PER-1-type extended-spectrum beta-lactamases among nosocomial Acinetobacter and Pseudomonas aeruginosa isolates in Turkey: a nationwide multicenter study. AB - We studied the prevalence and molecular epidemiology of PER-1-type beta lactamases among Acinetobacter, Klebsiella, and Pseudomonas aeruginosa strains isolated over a 3-month period in eight university hospitals from distinct regions of Turkey. A total of 72, 92, and 367 Acinetobacter, Klebsiella, and P. aeruginosa isolates were studied, respectively. The presence of blaPER was determined by the colony hybridization method and later confirmed by isoelectric focusing. We detected PER-1-type beta-lactamases in 46% (33/72) of Acinetobacter strains and in 11% (40/367) of P. aeruginosa strains but not in Klebsiella strains. PER-1-type enzyme producers were highly resistant to ceftazidime and gentamicin, intermediately resistant to amikacin, and susceptible or moderately susceptible to imipenem and meropenem. Among PER-1-type-beta-lactamase-positive isolates, five Acinetobacter isolates and six P. aeruginosa isolates from different hospitals were selected for ribosomal DNA fingerprinting with EcoRI and SalI. The EcoRI-digested DNAs were later hybridized with a digoxigenin-labelled PER-1 probe. The ribotypes and the lengths of blaPER-carrying fragments were identical in four Acinetobacter strains. A single isolate (Ac3) harbored a PER gene on a different fragment (approximately 4.2 kbp) than the others (approximately 3.4 kbp) and showed a clearly distinguishable ribotype. Ribotypes of P. aeruginosa strains obtained with EcoRI showed three patterns. Similarly, in Pseudomonas strains two different EcoRI fragments harbored blaPER (approximately 4.2 kbp in five isolates and 3.4 kbp in one isolate). PER-1-type beta-lactamases appear to be restricted to Turkey. However, their clonal diversity and high prevalence indicate a high spreading potential. PMID- 9333060 TI - Role of embB in natural and acquired resistance to ethambutol in mycobacteria. AB - The mycobacterial embCAB operon encodes arabinosyl transferases, putative targets of the antimycobacterial agent ethambutol (EMB). Mutations in embB lead to resistance to EMB in Mycobacterium tuberculosis. The basis for natural, intrinsic resistance to EMB in nontuberculous mycobacteria (NTM) is not known; neither is the practical implication of resistance to EMB in the absence of embB mutations in M. tuberculosis well understood. The conserved embB resistance-determining region (ERDR) of a collection of 13 strains of NTM and 12 EMB-resistant strains of M. tuberculosis was investigated. Genotypes were correlated with drug susceptibility phenotypes. High-level natural resistance to EMB (MIC, . or =64 microg/ml) was associated with a variant amino acid motif in the ERDR of M. abscessus, M. chelonae, and M. leprae. Transfer of the M. abscessus emb allele to M. smegmatis resulted in a 500-fold increase in the MICs. In M. tuberculosis, embB mutations were associated with MICs of > or =20 microg/ml while resistance not associated with an ERDR mutation generally resulted in MICs of < or =10 microg/ml. These data further support the notion that the emb region determines intrinsic and acquired resistance to EMB and might help in the reassessment of the current recommendations for the screening and treatment of infections with EMB-resistant M. tuberculosis and NTM. PMID- 9333061 TI - Absorption of ofloxacin isomers in the rat small intestine. AB - Ofloxacin, a chiral fluoroquinolone, possesses two optical isomers. The antibacterial activity of S-(-)-ofloxacin is 8 to 128 times higher than that of R (+)-ofloxacin. In the rat, a saturable absorption process has been described for racemic ofloxacin. In the present study we investigated the mechanism underlying the in vivo intestinal absorption of ofloxacin enantiomers in the rat. Blood samples were collected from the portal vein. Our results show that the intestinal absorption of ofloxacin isomers is pH dependent, both enantiomers being best absorbed at neutral pH. S-(-)-Ofloxacin seems to have a greater affinity for the intestinal transporter (initial concentrations at 5 min [C(init)] are 0.17 +/- 0.04 and 0.12 +/- 0.03 microg/ml for S-(-)- and R-(+)-ofloxacin, respectively). Dipeptides fail to modify ofloxacin absorption, but amino acids reduce both isomers' absorption (C(init) is reduced by 53 and 33% with glycine for S-(-)- and R-(+)-ofloxacin, respectively, and by 59 and 42% with L-leucine). Gamma amino butyric acid interferes with the absorption of ofloxacin isomers, but less seriously than do amino acids. Furthermore, ofloxacin competes with other fluoroquinolones or P-glycoprotein substrates for a common secretory pathway, resulting in an increased rate of absorption for both ofloxacin isomers; this is probably an indirect result of their reduced efflux from the apical side of intestinal cells. PMID- 9333062 TI - Therapeutic efficacy of BO-3482, a novel dithiocarbamate carbapenem, in mice infected with methicillin-resistant Staphylococcus aureus. AB - The in vivo activity of BO-3482, which has a dithiocarbamate chain at the C-2 position of 1beta-methyl-carbapenem, was compared with those of vancomycin and imipenem in murine models of septicemia and thigh infection with methicillin resistant Staphylococcus aureus (MRSA). Because BO-3482 was more susceptible than imipenem to renal dehydropeptidase I in a kinetic study of hydrolysis by this renal enzyme, the therapeutic efficacy of BO-3482 was determined during coadministration with cilastatin. In the septicemia models, which involved two homogeneous MRSA strains and one heterogeneous MRSA strain, the 50% effective doses were, respectively, 4.80, 6.06, and 0.46 mg/kg of body weight for BO-3482; 5.56, 2.15, and 1.79 mg/kg for vancomycin; and >200, >200, and 15.9 mg/kg for imipenem. BO-3482 was also as effective as vancomycin in an MRSA septicemia model with mice with cyclophosphamide-induced immunosuppression. In the thigh infection model with a homogeneous MRSA strain, the bacterial counts in tissues treated with BO-3482-cilastatin were significantly reduced in a dose-dependent manner compared with the counts in those treated with vancomycin and imipenem-cilastatin (P < 0.001). These results indicate that BO-3482-cilastatin is as effective as vancomycin in murine systemic infections and is more bactericidal than vancomycin in local-tissue infections. The potent in vivo activity of BO-3482-cilastatin against such MRSA infections can be ascribed to the good in vitro anti-MRSA activity and improved pharmacokinetics in mice when BO-3482 is combined with cilastatin and to the bactericidal nature of the carbapenem. PMID- 9333063 TI - In vitro evaluation of BO-3482, a novel dithiocarbamate carbapenem with activity against methicillin-resistant staphylococci. AB - BO-3482, a dithiocarbamate carbapenem, inhibited clinical isolates of methicillin resistant staphylococci (MRS) at 6.25 microg/ml (MIC at which 90% of isolates tested are inhibited [MIC90]), while the MIC90 of imipenem was > 100 microg/ml. BO-3482 was generally less active than imipenem against methicillin-susceptible Staphylococcus aureus, streptococci, enterococci, and gram-negative bacteria, although BO-3482 showed better activity (MIC90) than imipenem against Enterococcus faecium, Haemophilus influenzae, Proteus mirabilis, and Clostridium difficile. The affinities (50% inhibitory concentrations) of BO-3482 for penicillin-binding protein (PBP) PBP 2' of MRS and PBP 5 of E. faecium (both PBPs have low affinities for ordinary beta-lactam antibiotics) were 3.8 and 20 microg/ml, respectively, reflecting the greater activity of BO-3482 against MRS than against E. faecium. PMID- 9333064 TI - Allicin from garlic strongly inhibits cysteine proteinases and cytopathic effects of Entamoeba histolytica. AB - The ability of Entamoeba histolytica trophozoites to destroy monolayers of baby hamster kidney cells is inhibited by allicin, one of the active principles of garlic. Cysteine proteinases, an important contributor to amebic virulence, as well as alcohol dehydrogenase, are strongly inhibited by allicin. PMID- 9333065 TI - Mutations in the gyrA and parC genes in fluoroquinolone-resistant clinical isolates of Pseudomonas aeruginosa. AB - We determined partial sequences of the gyrA and parC genes of the fluoroquinolone susceptible strain ATCC 27853 and 22 clinical isolates of Pseudomonas aeruginosa. While a single amino acid change in GyrA with or without a change in ParC was found in 14 isolates with decreased susceptibility to fluoroquinolones, 3 higher level fluoroquinolone-resistant isolates had a double amino acid change in GyrA and a single amino acid change in ParC. PMID- 9333067 TI - Low-level release of Shiga-like toxin (verocytotoxin) and endotoxin from enterohemorrhagic Escherichia coli treated with imipenem. AB - Shiga-like toxin (SLT) and endotoxin may participate in the pathogenesis of enterohemorrhagic Escherichia coli (EHEC) infection. Levels of release of SLT and endotoxin from EHEC treated in vitro with antibiotics were estimated. There were differential levels of release of SLT and endotoxin from EHEC treated with different antibiotics. Treatment of EHEC strains, namely, E. coli O157, O111, and O26, with imipenem induced much lower levels of release of SLT and endotoxin than treatment with ceftazidime. PMID- 9333066 TI - Pharmacokinetic study of cefodizime and ceftriaxone in sera and bones of patients undergoing hip arthroplasty. AB - The study was carried out to determine the concentrations of cefodizime (single 2 g intravenous [i.v.] dose) and ceftriaxone (single 2-g i.v. dose) in the sera and bones of 42 patients (18 women and 24 men) undergoing hip arthroplasty. The concentrations of cefodizime and ceftriaxone in cancellous and cortical bone appear to be related to the free levels in serum but not to the total levels in serum, so the concentrations of cephalosporins in bone must be compared with the free concentrations in serum. Both drugs diffuse well into bone, with concentrations exceeding the MIC at which 90% of the major pathogenic infecting organisms are inhibited. PMID- 9333068 TI - In vitro activities of 10 antimicrobial agents against bacterial vaginosis associated anaerobic isolates from pregnant Japanese and Thai women. AB - The in vitro activities of 10 antimicrobial agents against 159 bacterial vaginosis-associated anaerobic isolates from pregnant Japanese and Thai women were determined. Clindamycin, imipenem, cefmetazole, amoxicillin, amoxicillin clavulanate, and metronidazole were highly active against all anaerobic isolates except Prevotella bivia and Mobiluncus species, which were resistant to amoxicillin and metronidazole, respectively. Cefotiam, ceftazidime, and ofloxacin were variably effective, while cefaclor was the least effective agent. PMID- 9333069 TI - In vitro activities of the biguanide PS-15 and its metabolite, WR99210, against cycloguanil-resistant Plasmodium falciparum isolates from Thailand. AB - The in vitro activities of the new biguanide PS-15 and its putative active metabolite, WR99210, were determined against seven different isolates or clones of Plasmodium falciparum. The mean 50% inhibitory concentrations of PS-15 and WR99210 were 1,015 and 0.06 ng/ml, respectively. WR99210 was up to 363 times more potent than cycloguanil, the active metabolite of proguanil, against cycloguanil resistant parasites. The pronounced activity of WR99210 against multidrug resistant P. falciparum indicates that further studies are required to determine the value of the prodrug, PS-15, as an antimalarial agent. PMID- 9333070 TI - Involvement of a 43-kilodalton outer membrane protein in beta-lactam resistance of Shigella dysenteriae. AB - A beta-lactam-sensitive strain (C152) of Shigella dysenteriae showed two major outer membrane proteins (OMPs) with M(r)s of 43,000 and 38,000, while the clinical isolate M2 lacked the 43,000-Mr OMP, which acted as a channel for beta lactam antibiotics. Permeability of beta-lactams across the outer membrane (OM) of M2 was lower than that across the OM of C152. Mutants deficient in the 43-kDa OMP could be selected in vitro from strain C152 in the presence of cefoxitin. All beta-lactam-resistant strains were sensitive to imipenem. PMID- 9333071 TI - In vitro antimalarial activity of penduline, a bisbenzylisoquinoline from Isopyrum thalictroides. AB - Two bisbenzylisoquinolines, tetrandrine and penduline, were purified from Isopyrum thalictroides. When tested for antimalarial activity in vitro, penduline was efficient at concentrations fivefold lower than those of tetrandrine. In highly synchronized parasite cultures, penduline mostly interfered between the 8th and the 32nd hours of the parasite cycle. PMID- 9333072 TI - Quinolone susceptibility of norA-disrupted Staphylococcus aureus. AB - The MIC of norfloxacin for the norA-disrupted mutant termed RDN1, obtained from quinolone-susceptible Staphylococcus aureus RN4220, was eightfold lower than that for RN4220. The increase in susceptibility was related to an increase of drug accumulation by RDN1. These results indicate that NorA plays an important role in the susceptibility of quinolone-susceptible S. aureus to selected quinolones. PMID- 9333073 TI - Activity of SCH 56592 compared with those of fluconazole and itraconazole against Candida spp. AB - The in vitro activity of Schering 56592, a new azole drug, was compared with those of fluconazole and itraconazole against 103 isolates of Candida comprising 10 different species. Schering 56592 was more active than itraconazole and fluconazole, and it was active against many fluconazole-resistant isolates. PMID- 9333074 TI - Comparative in vitro activities of trovafloxacin (CP-99,219) against 221 aerobic and 217 anaerobic bacteria isolated from patients with intra-abdominal infections. AB - Four hundred thirty-eight bacteria cultured from specimens of patients with serious intra-abdominal infections were tested by agar dilution against trovafloxacin and other quinolones and antimicrobial agents. Trovafloxacin inhibited 435 strains (99.3%) at < or =2 microg/ml. All the quinolones had similar activities against Enterobacteriaceae and Pseudomonas sp., but trovafloxacin showed superior activities against streptococci, enterococci, and anaerobic organisms. Because of its excellent in vitro activities against diverse bacteria, trovafloxacin has potential use as a single agent for polymicrobial infections. PMID- 9333075 TI - Pyronaridine for treatment of Plasmodium ovale and Plasmodium malariae infections. AB - The clinical efficacy of oral pyronaridine was assessed in 22 symptomatic Cameroonian patients infected with Plasmodium ovale or Plasmodium malariae. All patients were cured on or before day 4. In vitro drug assays confirmed the sensitivity of P. ovale and P. malariae isolates to chloroquine and pyronaridine. PMID- 9333076 TI - Survival of immunoglobulins from different species through the gastrointestinal tract in healthy adult volunteers: implications for human therapy. PMID- 9333077 TI - Quarantine changes take effect in Hawaii. PMID- 9333079 TI - Human drug product not equivalent to veterinary ceftiofur. PMID- 9333078 TI - Public health veterinarians protect food supply. PMID- 9333080 TI - Illinois, FDA issue lead products notice. PMID- 9333081 TI - Update on BSE contingency plan. PMID- 9333082 TI - Employer-based outcomes assessment of recent graduates and comparison with performance during veterinary school. PMID- 9333083 TI - What is your diagnosis? Well-circumscribed osseous or enamel opacities associated with the caudal root of the mandibular third premolar and rostral root of the fourth premolar. Odontoma. PMID- 9333084 TI - Contracts for continuing service--herd care. PMID- 9333085 TI - Cost-effectiveness analysis of treatment alternatives for beef bulls with preputial prolapse. AB - OBJECTIVE: To develop an economic model for comparing cost-effectiveness of medical and surgical treatment versus replacement of beef bulls with preputial prolapse. DESIGN: Economic analysis. SAMPLE POPULATION: Estimates determined from medical records of bulls treated for preputial prolapse at our hospital and from information about treatment of bulls published elsewhere. PROCEDURE: Annual depreciation cost for treatment (ADC(T)) and replacement (ADC(R)) were calculated. Total investment for an injured bull equaled the sum of salvage value, maintenance cost, and expected cost of the treatment option under consideration. Total investment for a replacement bull was purchase price. Net present value of cost was calculated for each year of bull use. Sensitivity analyses were constructed to determine the value that would warrant treatment of an injured bull. RESULTS: The decision to treat was indicated when ADC(T) was less than ADC(R). In our example, it was more cost-effective for owners to cull an injured bull. The ADC(R) was $97 less than ADC(T) for medical treatment ($365 vs $462) and $280 less than ADC(T) for surgical treatment ($365 vs $645). Likewise, net present value of cost values indicated that it was more cost effective for owners to cull an injured bull. Sensitivity analysis indicated treatment decisions were justified on the basis of replacement value or planned number of breeding seasons remaining for the bull. CLINICAL IMPLICATIONS: The model described here can be used by practitioners to provide an objective basis to guide decision making of owners who seek advice on whether to treat or replace bulls with preputial prolapse. PMID- 9333086 TI - Extralabel use of nonsteroidal anti-inflammatory drugs. PMID- 9333088 TI - Use of partial prostatectomy for treatment of prostatic abscesses and cysts in dogs. AB - OBJECTIVE: To determine whether dogs had prostatic disease, urinary incontinence, or urinary tract infection 1 year after partial prostatectomy to treat prostatic abscesses and cysts. DESIGN: Prospective study. ANIMALS: 20 male dogs with prostatic abscesses or cysts. Fifteen dogs had evidence of urinary tract infection. Only 8 dogs urinated normally; the remainder dribbled, had obstructions, or required medical treatment. PROCEDURE: Partial prostatectomy was performed on each dog. Sexually intact dogs (n = 12) also were castrated. RESULTS: None of the dogs had return of prostatic cystic enlargement or clinical signs of prostatic disease during the first year after surgery. Two dogs were euthanatized within 1 year after surgery, with 1 dog having prostatic enlargement and adenocarcinoma and 1 dog having unrelated lymphosarcoma. Fifteen dogs were continent. The remaining 5 dogs urinated normally but had intermittent and minor incontinence. Eleven dogs had no signs of infection 1 year after surgery, 5 had pyuria or positive urine bacteriologic culture results, 2 did not have urinalysis performed, and 2 were euthanatized. CLINICAL IMPLICATIONS: Dogs with severe prostatic abscesses or cysts and infections can be successfully treated by partial prostatectomy with an ultrasonic surgical aspirator and castration, resulting in long-term disease resolution. Although most dogs with severe prostatic disease do not urinate normally before surgery, nearly all dogs resume normal micturition after partial prostatectomy. Postoperative results of partial prostatectomy appear to be better than those of previous drainage techniques for treatment of prostatic cavitary disease. PMID- 9333087 TI - Effects of iatrogenic blood contamination on results of cerebrospinal fluid analysis in clinically normal dogs and dogs with neurologic disease. AB - OBJECTIVE: To examine the effects that iatrogenic blood contamination would have on total protein concentration and nucleated cell count in CSF from clinically normal dogs and dogs with neurologic disease. DESIGN: Case-control study. STUDY POPULATION: 53 dogs confirmed to have neurologic disease and 21 clinically normal dogs. PROCEDURE: CSF samples were obtained from the cerebellomedullary cistern or the lumbar portion of the subarachnoid space. Red blood and nucleated cell counts were determined, and protein concentration was measured. RESULTS: RBC count was not significantly correlated with nucleated cell count or protein concentration in clinically normal dogs or dogs with neurologic disease. CLINICAL IMPLICATIONS: High CSF nucleated cell counts and protein concentrations are indicative of neurologic disease, even if samples contain moderate amounts of blood contamination. PMID- 9333089 TI - Femoral artery occlusion in Cavalier King Charles Spaniels. AB - OBJECTIVE: To investigate development of femoral artery occlusion in Cavalier King Charles Spaniels. DESIGN: Prospective study. ANIMALS: 954 Cavalier King Charles Spaniels. PROCEDURE: 1,750 cardiovascular examinations consisting of visual inspection of mucous membranes, thoracic auscultation in areas associated with the heart valves, thoracic palpation, and palpation of the femoral arteries were made at 10 dog shows on 954 dogs. Findings of clinically normal, weak, or undetectable femoral pulses were recorded. Pathologic changes in occluded femoral arteries of 2 dogs were examined histologically. RESULTS: Of the 954 dogs, 22 (2.3%) had an undetectable right or left femoral pulse on 1 or more examinations. Forty (4.2%) additional dogs had weak unilateral or bilateral femoral pulses. Only 1 dog had exercise intolerance, and it had coexistent congestive heart failure. Histologic examination of serial sections of an occluded femoral artery from 1 dog revealed intimal thickening with breaks in the internal elastic lamina proximal to the occluded segment. The occluded segment of the femoral artery was contracted and filled with an organizing, recanalizing thrombus. Similar histopathologic changes were found in sections of a femoral artery from another dog. CLINICAL IMPLICATIONS: Femoral artery occlusion is rare in other breeds and is not clinically important in dogs because of adequate collateral circulation; however, its rather common development in Cavalier King Charles Spaniels indicates a genetic predisposition and probable weakness in the femoral artery wall. PMID- 9333090 TI - Management of a congenitally shortened soft palate in a dog. AB - An 8-week-old puppy that was examined because of a nasal discharge was found to have a congenitally shortened soft palate. The palate was partially rebuilt with pharyngeal flaps constructed from the tonsillar crypts. The reconstructed soft palate extended approximately 40% of the distance between the caudal aspect of the hard palate and rostral tip of the epiglottis. During the first 18 months after surgery, the dog had 3 episodes of halitosis, sneezing, and mucopurulent nasal discharge but responded to antimicrobial treatment. The dog was fed dry dog food and drank water from an elevated bowl. Water would flow from the dog's nose if it drank water with its head lowered. Compensatory mechanisms likely play an important role in the outcome of animals with this condition. PMID- 9333091 TI - Malignant colonic neoplasia in cats: 46 cases (1990-1996). AB - OBJECTIVE: To evaluate a group of cats with malignant colonic neoplasia and to identify factors related to survival time. DESIGN: Retrospective study. ANIMALS: 46 cats with malignant colonic neoplasia. PROCEDURE: Information on signalment, diagnostic findings, histopathologic diagnosis, surgical procedure performed, identification of nodal metastasis at surgery, type of chemotherapy administered, and survival time was obtained from the medical record of each cat. A diagnosis of malignant colonic neoplasia had been established by histologic examination of endoscopic biopsy specimens (3 cats), biopsy specimens obtained during laparotomy (38), or necropsy specimens (5). RESULTS: Mean age of cats was 12.5 years (range, 6 to 18 years). Ultrasonography was useful 84% of the time in localizing the mass to the intestine. Three cats had endoscopic biopsy, 9 had incisional biopsy, 21 had mass resection, and 8 had subtotal colectomy performed. Histopathologic diagnoses included adenocarcinoma (21 cats), lymphoma (19), mast cell tumor (4), and neuroendocrine carcinoma (2). CLINICAL IMPLICATIONS: Obtaining clean margins at surgery seems to increase survival time in cats with malignant colonic neoplasia. Metastasis at the time of surgery decreases survival time. Data from this study indicate that the survival time of certain cats with colonic lymphoma may not be affected by chemotherapy. Cats with an unidentified colonic mass should receive a subtotal colectomy to increase survival time. Cats with colonic adenocarcinoma should receive a subtotal colectomy with consideration of doxorubicin administration to increase survival time. PMID- 9333093 TI - Influence of preoperative complete blood cell counts on surgical outcomes in healthy horses: 102 cases (1986-1996). AB - OBJECTIVE: To assess the value of CBC as a preoperative test in healthy horses undergoing cryptorchidectomy. DESIGN: Retrospective study. ANIMALS: 117 horses. PROCEDURE: Medical records were reviewed to identify horses that had had cryptorchidectomy. Of the 117 horses identified, 102 were found that did not have a known medical condition believed to adversely affect surgery. Preoperative CBC was assessed in terms of abnormalities detected and effects of these abnormalities on development of complications during and after surgery and patient management. RESULTS: Of 102 CBC performed, 55 contained abnormalities. Mild neutrophilia was detected in 40 horses. Of the remaining 15 horses with abnormal CBC, 8 had abnormalities that were considered potentially important. Changes in patient management and development of surgical complications were not associated with these 8 horses. Surgical complications were defined as intraoperative changes in blood pressure, excessive blood loss, development of postanesthetic myopathies, and postoperative wound or respiratory tract infections. Surgical complications developed in 17 horses. Of these, 6 horses had an abnormal CBC that was indicative of mild neutrophilia. Positive and negative predictive values of preoperative CBC on development of perioperative complications were 0.11 and 0.77, respectively. CLINICAL IMPLICATIONS: On the basis of findings in our study, determination of preoperative CBC does not predict development of complications during or after surgery and does not alter patient management. PMID- 9333092 TI - Brodifacoum toxicosis in two horses. AB - Increased popularity during the past decade of brodifacoum, an anticoagulant rodenticide, has led to an increase in cases of accidental poisoning in nontarget species, including pets and farm animals. Pharmacokinetics of second-generation anticoagulant rodenticides such as brodifacoum are substantially different from those of first-generation anticoagulant rodenticides such as warfarin. This difference dramatically influences management of exposure in terms of duration and cost of treatment and may affect outcome. The National Poison Control Center reports that approximately 50 cases of brodifacoum exposure have occurred in horses between 1993 and 1997. To our knowledge, this report is the first complete clinical description of accidental ingestion of a potentially lethal dose of brodifacoum in horses. Early recognition of exposure to brodifacoum, subsequent treatment with adequate doses of vitamin K1, and sequential monitoring of clotting times and serum brodifacoum concentration permitted poisoning in these horses to be managed successfully. PMID- 9333094 TI - Use of antibiotic-impregnated polymethyl methacrylate in horses with open or infected fractures or joints: 19 cases (1987-1995). AB - OBJECTIVE: To evaluate the clinical efficacy of antibiotic-impregnated polymethyl methacrylate (PMMA) in horses with open or infected fractures or joints in which internal fixation or external coaptation devices were used. DESIGN: Retrospective case series. ANIMALS: 19 horses in which antibiotic-impregnated PMMA was used as part of the treatment regimen. PROCEDURES: Medical records of each horse were reviewed, and owners and trainers were contacted to provide additional information. RESULTS: Musculoskeletal problems in these horses included 10 fractures of long bones, 2 comminuted phalangeal fractures, 5 joint injuries, and 2 chronically septic joints in which ankylosis was stimulated. Nine horses had open fractures, 8 had closed wounds and developed infection after internal fixation of fractures, and 2 had chronically septic joints. Bony union was achieved in 15 of 19 horses. Twelve horses were discharged from the hospital and survived long term. Gentamicin sulfate, tobramycin sulfate, amikacin sulfate, and cefazolin sodium were used in PMMA. CLINICAL IMPLICATIONS: Use of antibiotic impregnated PMMA provided high local concentrations of antibiotics and should be considered in the treatment of horses with open fractures and acute and chronic bone and joint infections. PMID- 9333095 TI - Factors associated with prolonged weaning-to-mating interval among sows on farms that wean pigs early. AB - OBJECTIVE: To determine factors associated with weaning-to-mating interval among sows on commercial farms that wean pigs early. DESIGN: Cohort study. ANIMALS: 11,861 farrowing sows. PROCEDURE: Production, farrowing, and feed intake records were reviewed for sows on 16 farms for which mean duration of lactation was between 14.9 and 18.9 days. RESULTS: Among sows with high feed intake during lactation (> or = 5.6 kg/d [12.3 lb/d]), lactation duration was not associated with weaning-to-mating interval, but among sows with low feed intake during lactation (< 5.6 kg/d), weaning-to-mating interval increased as lactation duration decreased. Furthermore, among sows with the lowest feed intake during lactation (< 4.2 kg/d [9.2 lb/d]), those that had heavier litter weights at weaning (> 54 kg [119 lb]) had a longer weaning-to-mating interval than did those that had lighter litter weights at weaning. Sows with low feed intake and high litter weight at weaning accounted for 5 to 20% of sows on each farm. In general, weaning-to-mating interval increased as parity decreased, but the change in weaning-to-mating interval associated with a particular change in lactation duration varied with parity. CLINICAL IMPLICATIONS: When feed intake during lactation is maximized (> or = 5.6 kg/d), lactation duration is not significantly associated with weaning-to-mating interval. Producers should consider fostering or partially weaning litters when sows with high litter weights are not consuming sufficient feed. PMID- 9333096 TI - Cholelithiasis and cholecystitis in a dairy cow. AB - A 9-year-old Holstein cow was evaluated for colic and decreased milk production of 2 days' duration. Preoperative serum biochemical results suggested hepatic damage and cholestasis. On the basis of persistent signs of abdominal pain that were nonresponsive to analgesics, exploratory laparotomy was performed. The cow was found to have choleliths. Cholecystocentesis was performed, and samples were submitted for cytologic examination and bacterial culture. Bacterial culture yielded Escherichia coli and Clostridium perfringens. Using digital pressure, choleliths were reduced until they could be passed through the bile duct into the duodenum. The cow recovered from surgery without complications, and all serum biochemical test results returned to reference ranges. Cholelithiasis is rare in cattle but can result in signs of abdominal pain. PMID- 9333097 TI - Bronchoalveolar lavage in a dolphin. AB - Bronchoalveolar lavage (BAL) was performed twice to evaluate a stranded Atlantic bottle-nosed dolphin for pulmonary disease. A pediatric gastroscope with a working length of 1,090 mm and an outer diameter of 9.8 mm was of appropriate size for BAL in this dolphin. Fifty milliliters of sterile saline (0.9% NaCl) solution was used in each of 2 sites for the first lavage. Fluid recovery was 58 and 66% from the 2 sites; however, results of cytologic analyses were typical of a bronchial wash rather than BAL. Larger volumes of saline solution (85 to 100 ml/site) were used in the second lavage. Although fluid recovery was only 25 and 30% from the 2 sites, results of cytologic analyses were consistent with BAL. Mononuclear cells accounted for 72 and 90% of total WBC. Although the dolphin of this report did not appear to have pulmonary disease, experience obtained by performing BAL provided valuable information for the practical application of this technique in dolphins. PMID- 9333098 TI - Clinical pharmacology education in primary care residency programs: Education Committee, American Society for Clinical Pharmacology and Therapeutics. PMID- 9333099 TI - Issues in pharmaceutical lotteries: the case of interferon beta-1b. PMID- 9333100 TI - Role of intestinal P-glycoprotein (mdr1) in interpatient variation in the oral bioavailability of cyclosporine. AB - Interpatient differences in the oral clearance of cyclosporine (INN, ciclosporin) have been partially attributed to variation in the activity of a single liver enzyme termed CYP3A4. Recently it has been shown that small bowel also contains CYP3A4, as well as P-glycoprotein, a protein able to transport cyclosporine. To assess the importance of these intestinal proteins, the oral pharmacokinetics of cyclosporine were measured in 25 kidney transplant recipients who each had their liver CYP3A4 activity quantitated by the intravenous [14C-N-methyl]-erythromycin breath test and who underwent small bowel biopsy for measurement of CYP3A4 and P glycoprotein. Forward multiple regression revealed that 56% (i.e., r2 = 0.56) and 17% of the variability in apparent oral clearance [log (dose/area under the curve)] were accounted for by variation in liver CYP3A4 activity (p < 0.0001) and intestinal P-glycoprotein concentration (p = 0.0059), respectively. For peak blood concentration, liver CYP3A4 activity accounted for 32% (p = 0.0002) and P glycoprotein accounted for an additional 30% (p = 0.0024) of the variability. Intestinal levels of CYP3A4, which varied tenfold, did not appear to influence any cyclosporine pharmacokinetic parameter examined. We conclude that intestinal P-glycoprotein plays a significant role in the first-pass elimination of cyclosporine, presumably by being a rate-limiting step in absorption. Drug interactions with cyclosporine previously ascribed to intestinal CYP3A4 may instead be mediated by interactions with intestinal P-glycoprotein. PMID- 9333102 TI - Pharmacokinetics of valsartan in patients with liver disease. AB - OBJECTIVES: Valsartan (CGP 48933), an orally active angiotensin II antagonist, is eliminated mainly by hepatic clearance. To characterize the compound(s) excreted in the bile, biliary excretion of valsartan was investigated by collection of bile after an intravenous dose of valsartan. In addition, to determine the exposure to valsartan when liver function is impaired, a pharmacokinetic study (open, single dose) was performed in patients with mild and moderate impairment of liver function. PATIENTS: Biliary excretion of valsartan (after intravenous administration of 20 mg valsartan) was assessed in a patient who underwent a hepaticojejunostomy with subsequent bile drainage. Exposure to valsartan in patients with mild (n = 6) or moderate (n = 6) impaired liver function (Child's Pugh classification) and matching (sex, age, and weight) healthy volunteers (n = 12) was studied after oral administration of a single dose of 160 mg valsartan. RESULTS: After intravenous administration, valsartan was eliminated mainly as unchanged drug in the bile. Mean exposure (measured as area under the plasma valsartan concentration-time curve) to valsartan was increased about twofold in both the mild and the moderate groups compared with matched (age, sex, and weight) healthy volunteers. CONCLUSION: These data are consistent with the pharmacokinetics of valsartan in that biliary excretion is the main route of elimination. PMID- 9333101 TI - N-acetylation among HIV-positive patients and patients with AIDS: when is fast, fast and slow, slow? AB - BACKGROUND: The discrepancy between genotype and expressed phenotype of the polymorphic N-acetyltransferase (NAT2) has been suggested by separate genotypic and phenotypic studies in populations with human immunodeficiency virus (HIV). Only one study has examined both genotype and phenotype in the same population, and no discrepancies were observed. METHODS: In a cross-sectional study, 105 HIV positive patients and patients with acquired immunodeficiency syndrome (AIDS) were phenotyped for NAT2 activity with use of caffeine as an in vivo probe; 50 of these patients were also genotyped by restriction mapping and allele-specific amplification. In a longitudinal study, 23 patients were phenotyped at least twice during the 2-year study. RESULTS: The distribution of the NAT2 phenotype among the 105 patients was unimodal and skewed toward slow acetylators as opposed to the bimodal distribution observed in healthy white populations. The genotype distribution was 26:24 slow:fast. There were 18 discrepancies between genotype and phenotype: 12 slow acetylators with fast genotypes and six fast acetylators with slow genotypes. No drug-related effects on NAT2 activity were apparent, but the role of disease progression was evident. Among the slow acetylators whose genotype was fast, the incidence of AIDS was higher (six of 12) than that among the fast acetylators whose genotype was fast (two of 14). Among patients phenotyped more than once (mean time between samples, 10.4 months) changes in phenotype from fast to slow were associated with progression of HIV infection. CONCLUSIONS: Disease progression in HIV infection and AIDS may alter expression of the NAT2 gene. The genotype and the phenotype are not interchangeable measurements. In the HIV population, to know the genotype is useful only if the phenotype is also known and vice versa. PMID- 9333103 TI - Fluvoxamine inhibits the CYP2C19-catalyzed bioactivation of chloroguanide. AB - OBJECTIVE: To investigate the interaction between fluvoxamine and chloroguanide (INN, proguanil) to confirm that fluvoxamine inhibits CYP2C19. METHODS: The study was carried out with a randomized, in vivo, crossover design. Six volunteers were extensive metabolizers of the S-mephenytoin oxidation polymorphism, and six volunteers were poor metabolizers. In period A of the study, each subject took 200 mg chloroguanide orally. In period B, each subject took 100 mg/day fluvoxamine for 8 days and on day 6 ingested 200 mg chloroguanide. In both periods, blood and urine were sampled at regular intervals. Chloroguanide and its two metabolites cycloguanil and 4-chlorphenylbiguanide in plasma and in urine were assayed by means of HPLC. RESULTS: During fluvoxamine use, the median of the total clearance of chloroguanide decreased in a statistically significant way from 1282 ml/min to 782 ml/min among the extensive metabolizers, whereas there was no change among the poor metabolizers. The partial clearance of chloroguanide by means of cydoguanil and 4-chlorphenylbiguanide formation among the extensive metabolizers decreased from 222 ml/min and 97 ml/min before to 33 ml/min and 11 ml/min during fluvoxamine intake, respectively. Among poor metabolizers the corresponding values were 35 ml/min and 7.6 ml/min before and 38 ml/min and 6.9 ml/min during fluvoxamine intake. For each metabolite clearance the change was statistically significant among the extensive metabolizers but not among the poor metabolizers. Both cycloguanil and 4-chlorphenylbiguanide formation clearances were statistically significantly higher among the extensive metabolizers than the poor metabolizers in period A but not in period B (phenocopy). CONCLUSION: Fluvoxamine is an effective inhibitor of CYP2C19. PMID- 9333105 TI - Diclofenac concentrations in defined tissue layers after topical administration. AB - INTRODUCTION: To date it is unclear whether therapeutic concentrations are attained in target tissues after topical administration of nonsteroidal anti inflammatory drugs. Therefore this study in healthy volunteers was undertaken to measure diclofenac concentrations attained in defined tissue layers directly underlying the site of topical diclofenac application by in vivo microdialysis. METHODS: In each experiment two microdialysis probes were inserted, one into a superficial (3.9 +/- 0.3 mm) and one into a deep (9.3 +/- 0.5 mm) tissue layer, in 20 healthy volunteers and calibrated in vivo. The distance between the surface of the skin and the tips of the microdialysis probes was measured by 7.5 MHz ultrasound. Diclofenac was administered topically as a single dose of approximately 300 mg/100 cm2. Concentration versus time profiles in tissue layers were monitored for 5 hours. RESULTS: Concentration versus time profiles were obtained in 11 of 20 experiments. However, there was no correlation between area under the concentration-time curve (AUC) in a defined layer and the depth of probe insertion. In those experiments where concentration versus time profiles were obtained for both probes mean AUC was 532 +/- 197 microg x min x ml(-1) for superficial layers, and 438 +/- 249 microg x min x ml(-1) for deep layers. CONCLUSION: We conclude that transdermal penetration of diclofenac, at least after single doses, is not predictable and may strongly be influenced by individual skin properties. PMID- 9333104 TI - Genetic polymorphism of the CYP2C subfamily and its effect on the pharmacokinetics of phenytoin in Japanese patients with epilepsy. AB - OBJECTIVE: To examine the genetic polymorphism of CYP2C9 and CYP2C19 and its effect on the pharmacokinetics of phenytoin among 44 Japanese patients with epilepsy. METHODS: Polymerase chain reaction tests with leukocyte deoxyribonucleic acid were used to detect the mutations for the amino acid substitution (Arg144-->Cys and Ile359-->Leu) in CgammaP2C9 and for the defective allele (m1 and m2) in CgammaP2C19. The pharmacokinetic parameters of phenytoin in individual patients were estimated by means of empirical bayesian analysis, in which the prior information was the population parameters for Japanese patients with epilepsy. RESULTS: Of the 44 patients, none had the CgammaP2C9 mutation for the Cys144 allele, whereas six patients were heterozygous for the wild-type (wt) and Leu359 allele (wt/Leu359) in cgammaP2C9. The maximal elimination rate (Vmax) of phenytoin among patients with heterozygous wt/Leu359 in CgammaP2C9 was 33% lower than that among patients with normal CgammaP2C9. A total of 21 patients were heterozygous for the CgammaP2C19 mutation (wt/m1 or wt/m2), and five patients had the homozygous or heterozygous mutations in CgammaP2C19 (m1/m1 or m1/m2). The Vmax values of phenytoin were slightly decreased (up to 14%) among patients with CgammaP2C19 mutations compared with patients with normal CgammaP2C19. CONCLUSION: The findings indicated that the genetic polymorphisms of CYP2C isozymes play an important role in the pharmacokinetic variability of phenytoin and that the mutation in CYP2C9 proteins (Ile359-->Leu) is a determinant of impaired metabolism of the drug among Japanese persons. PMID- 9333106 TI - Optimizing levodopa pharmacokinetics with multiple tolcapone doses in the elderly. AB - OBJECTIVES: The multiple-dose tolerability, pharmacokinetics, and pharmacodynamics of tolcapone, a novel catechol-O-methyltransferase (COMT) inhibitor, were assessed in healthy elderly volunteers receiving concomitant carbidopa and levodopa. METHODS: Thirty-six volunteers from 55 to 75 years old participated in this double-blind, placebo-controlled, ascending multiple-dose study. Tolcapone was studied at dosages of 100, 200, 400, or 800 mg three times daily (t.i.d.) in four sequential groups. Each group consisted of nine participants who had been randomized to receive either placebo (n = 3) or tolcapone (n = 6). Tolcapone or placebo was coadministered with carbidopa and levodopa (25 and 100 mg, respectively) for 7 days. Assessments included tolerability, pharmacokinetics of tolcapone, levodopa, and 3-O-methyldopa, and inhibition of COMT activity in erythrocytes. RESULTS: By inhibiting COMT, tolcapone reduced levodopa metabolism to 3-O-methyldopa, resulting in a twofold increase in levodopa exposure (area under the curve) and elimination half-life, without changing levodopa peak plasma concentration. These effects were similar on days 1 and 7 of treatment. Development of tolerance to COMT inhibition was not observed. Onset of effect was rapid (day 1 of treatment), and the maximum effect on levodopa pharmacokinetics was already observed with 100 or 200 mg tolcapone t.i.d. At these dosages, tolcapone pharmacokinetics were linear and stable; accumulation occurred with 800 mg t.i.d. The combination of tolcapone and carbidopa-levodopa was generally well tolerated, although more nausea and vomiting were observed at higher dosages (400 to 800 mg t.i.d.), particularly in women. CONCLUSION: Tolcapone shows promise as an effective adjunct to levodopa in the treatment of Parkinson's disease. Clinical pharmacology data indicate that the therapeutic regimen should be 100 or 200 mg t.i.d. PMID- 9333107 TI - Accumulation of lovastatin, but not pravastatin, in the blood of cyclosporine treated kidney graft patients after multiple doses. AB - OBJECTIVES: To study pravastatin and lovastatin pharmacokinetic and pharmacodynamic effects and their interactions with cydosporine (INN, ciclosporin) in kidney transplant patients after single and multiple doses. SUBJECTS AND METHODS: The pharmacokinetic and pharmacodynamic effects of administration of 20 mg/day oral pravastatin and lovastatin for 28 days and their interactions with cyclosporine (2 to 6 mg/kg/day) were studied in a double-blind, double-dummy, randomized, parallel-group multicenter trial in 44 stable kidney graft recipients. RESULTS: The median area under the curve [AUC(0-24)] of pravastatin was 249 microg x hr/L (range, 104 to 1026 microg x hr/L) after a single dose (day 1) and 241 microg x hr/L (114 to 969 microg x hr/L) after multiple doses (day 28) and was fivefold higher than values reported in the absence of cyclosporine. The median AUC(0-24) of lovastatin was 243 microg x hr/L (105 to 858 microg x hr/L) on day 1 and 459 microg x hr/L (140 to 1508 microg x hr/L) on day 28. Besides a significant accumulation during the study period (p < 0.001), the lovastatin AUC(0-24) values were twentyfold higher than values reported without cyclosporine. Coadministration of pravastatin or lovastatin did not alter cyclosporine pharmacokinetics. In this study, 20 mg/day doses of both drugs resulted in a significant improvement of the lipid profile and were well tolerated. CONCLUSIONS: In contrast to lovastatin, pravastatin did not accumulate over the study period, which is probably one of the reasons rhabdomyolysis has been reported in lovastatin-treated but not pravastatin-treated transplant patients receiving cyclosporine immunosuppression. PMID- 9333108 TI - Familial studies of heritability of alpha1-adrenergic receptor responsiveness in superficial veins. AB - BACKGROUND: Studies of monozygotic and dizygotic twins indicate an important genetic influence on the variability of responsiveness to norepinephrine in superficial human vein. OBJECTIVES: Genetic aspects of variability of alpha1 adrenergic receptor responsiveness to norepinephrine in superficial veins were further investigated by studying the response to norepinephrine in the dorsal hand veins of parents and their children. METHODS: Subjects were healthy nonsmoking adults (n = 24; age range, 40 to 52 years) and their biological children (n = 20; age range, 15 to 26 years) who were free from medications likely to modify vascular tone. Superficial vein responsiveness to norepinephrine was assessed by the linear variable differential transformer technique. The dose of norepinephrine required to constrict superficial vein diameter by 50% from baseline (ED50) was calculated for each subject. Heritability was estimated by standard techniques of regression of mid-parent/child (natural logarithm) ED50 values. RESULTS: ED50 ranged from 5.6 to 254.6 ng/min in the parents and from 7.8 to 242.3 ng/min in the children. Heritability was calculated at 0.88. CONCLUSIONS: These data confirm wide variability in superficial vein responsiveness to norepinephrine. The results confirm a major genetic influence in biological responsiveness of superficial vein to norepinephrine in healthy humans. Heritability of vascular alpha-adrenergic receptor responsiveness may influence vascular regulation during sympathetic stimulation and blockade. PMID- 9333109 TI - Endogenous angiotensin II does not contribute to sympathetic venoconstriction in dorsal hand veins of healthy humans. AB - BACKGROUND: Sympathetically mediated venoconstriction is augmented by exogenously administered angiotensin II. This study was designed to assess whether endogenous angiotensin II influences sympathetically mediated venous tone. METHODS: Responses of dorsal hand veins to local intravenous administration of subsystemic doses of losartan, an angiotensin II type-1 receptor antagonist, were assessed with use of a well-validated displacement technique in eight healthy male volunteers. In a four-phase study, responses to local infusions of angiotensin II (4 to 64 ng/min) and norepinephrine (1 to 128 ng/min) or to sympathetic venoconstriction produced by a single deep breath were compared in the presence of either saline placebo or 30 microg/min losartan. Each phase of the study was conducted on a separate day, in random order, and each phase was separated by at least 1 week. RESULTS: Angiotensin II (p = 0.03) and norepinephrine (p < 0.001) caused dose-dependent venoconstriction. Losartan attenuated the venoconstriction induced by angiotensin II (p = 0.048) but had no effect on the responses to norepinephrine or the venoconstriction induced by a single deep breath. CONCLUSIONS: In contrast to exogenously administered angiotensin II, basal endogenous angiotensin II does not influence sympathetically mediated venoconstriction in healthy humans. However, endogenous angiotensin II may have a role in circumstances of renin-angiotensin system activation, such as salt depletion. PMID- 9333110 TI - Paroxetine potentiates the central nervous system side effects of perphenazine: contribution of cytochrome P4502D6 inhibition in vivo. AB - BACKGROUND: Paroxetine is a frequently used antidepressant and a potent inhibitor of the CYP2D6 isozyme in vitro (inhibition constant [Ki] = 0.15 micromol/L). Most classic antipsychotic agents such as perphenazine are metabolized by the CYP2D6 isozyme and are often coadministered with antidepressant agents. This study assessed the extent of changes in CYP2D6 isozyme activity in vivo after pretreatment with paroxetine and its consequences on perphenazine kinetics and central nervous system effects. METHODS: Eight extensive metabolizers for CYP2D6 were administered a single dose of perphenazine (0.11 mg/kg orally) or placebo following a randomized double-blind design. Perphenazine plasma concentrations and effects were assessed for a period of 8 hours. Subsequently, subjects were treated with a standard therapeutic dose of paroxetine (20 mg/day orally) for 10 days and test sessions with perphenazine and placebo were repeated. RESULTS: Paroxetine treatment resulted in a twofold to 21-fold decrease in CYP2D6 activity (p < 0.001). After pretreatment with paroxetine, perphenazine peak plasma concentrations increased twofold to 13-fold (p < 0.01). This was associated with a significant increase in central nervous system side effects of perphenazine, including oversedation, extrapyramidal symptoms, and impairment of psychomotor performance and memory (p < 0.05). CONCLUSION: Coadministration of perphenazine after pretreatment with a standard therapeutic dose of paroxetine increased the plasma concentration and central nervous system side effects of perphenazine, primarily as a result of inhibition of the CYP2D6 isozyme. In patients who are at steady state with paroxetine, a reduction of perphenazine dose may be required to prevent central nervous system side effects. PMID- 9333111 TI - Plasma buspirone concentrations are greatly increased by erythromycin and itraconazole. AB - BACKGROUND: The oral bioavailability of buspirone is very low as a result of extensive first-pass metabolism. Erythromycin and itraconazole are potent inhibitors of CYP3A4, and they increase plasma concentrations and effects of certain drugs, for example, oral midazolam and triazolam. The possible interactions of buspirone with erythromycin and itraconazole have not been studied before. METHODS: The pharmacokinetics and pharmacodynamics of buspirone were investigated in a randomized, double-blind, double-dummy crossover study with three phases. Eight young healthy volunteers took either 1.5 gm/day erythromycin, 200 mg/day itraconazole, or placebo orally for 4 days. On day 4, 10 mg buspirone was administered orally. Timed blood samples were collected up to 18 hours, and the effects of buspirone were measured with four psychomotor tests up to 8 hours. RESULTS: Erythromycin and itraconazole increased the mean area under the plasma concentration-time curve from time zero to infinity [AUC(0-infinity] of buspirone about sixfold (p < 0.05) and 19-fold (p < 0.01), respectively, compared with placebo. The mean peak plasma concentration (Cmax) of buspirone was increased about fivefold (p < 0.01) and 13-fold (p < 0.01) by erythromycin and itraconazole, respectively. These interactions were evident in each subject, although a striking interindividual variability in the extent of both interactions was observed. The elimination half-life of buspirone did not seem to be prolonged by either erythromycin or itraconazole. The effect of itraconazole on the Cmax and AUC(0-infinity) of buspirone was significantly (p < 0.01) greater than that of erythromycin. The greatly elevated plasma buspirone concentrations resulted in increased (p < 0.05) pharmacodynamic effects (as measured by the Digit Symbol Substitution test and the Critical Flicker Fusion test) and in side effects of buspirone. CONCLUSIONS: Both erythromycin and itraconazole greatly increased plasma buspirone concentrations, obviously by inhibiting its CYP3A4 mediated first-pass metabolism. These pharmacokinetic interactions were accompanied by impairment of psychomotor performance and side effects of buspirone. The dose of buspirone should be greatly reduced during concomitant treatment with erythromycin, itraconazole, or other potent inhibitors of CYP3A4. PMID- 9333112 TI - Self-medication. PMID- 9333113 TI - On our age-related bone loss: insights from a new paradigm. AB - Bone strength and "mass" normally adapt to the largest voluntary loads on bones. The loads come from muscles, not body weight. Bone modeling can increase bone strength and "mass," bone remodeling can conserve or reduce them, and each can turn ON and OFF in response to its own threshold range of bone strains. During growth, the loads on bones from body weight and muscle forces increase, and modeling correspondingly increases bone strength and "mass." In young adults those loads usually plateau, so bone strength can "catch up" and modeling can turn OFF. Meanwhile remodeling keeps existing bone. After about 30 years of age, muscle strength usually decreases. In aging adults this would put bones that had adapted to stronger young-adult muscles into partial disuse and make remodeling begin to reduce their strength and "mass," as disuse regularly does in experimental situations in other mammals, both growing and adult. Those changes associate strongly with the size of the bone strains caused by the loads on bone. While nonmechanical effects associated with aging should contribute to that age related bone loss too, a new skeletal paradigm suggests the above mechanical influences would dominate control of the process in time and anatomical space. PMID- 9333114 TI - Muscle strength, bone mass, and age-related bone loss. PMID- 9333115 TI - The 25-hydroxyvitamin D 1-alpha-hydroxylase gene maps to the pseudovitamin D deficiency rickets (PDDR) disease locus. AB - Pseudovitamin D-deficiency rickets (PDDR) is an autosomal recessive disorder that may be due to impaired activity of 25-hydroxyvitamin D-1alpha-hydroxylase, a renal cytochrome P450 enzyme (P450[1alpha]) of the vitamin D pathway. The disease locus for PDDR has been mapped by linkage analysis to 12q13-q14, but the molecular defect underlying the enzyme dysfunction has remained elusive due to the lack of sequence information for the P450(1alpha) gene (hereafter referred to as 1alpha-OHase). We have used a probe derived from the rat 25-hydroxyvitamin D 24-hydroxylase (CYP24; 24-OHase) sequence to identify and clone the 1alpha-OHase cDNA. The full-length 1alpha-OHase clone of 2.4 kb codes for a protein of predicted Mr 55 kDa. Functional activity of the cloned sequence was assessed using transient transfection, and the production of authentic 1alpha,25 dihydroxyvitamin D3 [1alpha,25(OH)2D3] was confirmed using high performance liquid chromatography fractionation and time-of-flight mass spectrometry. The expression of the gene was analyzed in vitamin D-replete animals; treatment with 1alpha,25(OH)2D3 reduced 1alpha-OHase transcript levels by 70%, while administration of parathyroid hormone led to a 2-fold increase in the expression of the gene, thus confirming the hormonal regulation previously described using biochemical methods. The rat cDNA was used to obtain a human genomic clone. Interestingly, the human 1alpha-OHase gene mapped to 12q13.1-q13.3, providing strong evidence that a mutation in the 1alpha-OHase gene is responsible for the PDDR phenotype. The availability of a cloned sequence for 1alpha-OHase generates novel tools for the study of the molecular etiology of PDDR, and will allow the investigation of other disturbances of vitamin D metabolism. PMID- 9333116 TI - Interleukin-1alpha and beta in growth plate cartilage are regulated by vitamin D metabolites in vivo. AB - Matrix remodeling plays a prominent role in growth plate calcification. Since interleukin-1 (IL-1) has been implicated in stimulating proteinase production and inhibiting matrix synthesis in articular cartilage, we examined whether IL-1 was present in growth plate and whether the vitamin D metabolites, 1,25 dihydroxyvitamin D3 (1,25(OH)2D3; 1,25) and 24,25(OH)2D3 (24,25), regulate the level of IL-1 found in this tissue. Sprague-Dawley rats were placed on normal (Normal rats) or rachitogenic diet (-VDP rats). The -VDP rats were either left untreated, injected 24 h prior to euthanasia with 24,25 (-VDP+24,25 rats) or 1,25 (-VDP+1,25 rats), or were given ergocalciferol (Ergo rats) orally, 48 h prior to euthanasia. Growth plates were harvested and extracted in buffer containing 1 M guanidine. IL-1 activity was measured by adding authentic cytokine or growth plate extracts to cultures of lapine articular cartilage and assaying release of glycosaminoglycans (GAGs) and changes in collagenase and neutral metalloproteinase activity. Neutralization of activity in the extracts was performed using polyclonal antisera to IL-1alpha or IL-1beta. An ELISA was used to determine levels of IL-1alpha and beta in the extracts. All extracts contained IL-1alpha and beta, as determined by ELISA. Levels of IL-1beta, but not IL 1alpha, were affected by the vitamin D status of the animal. Extracts from VDP+24,25 animals contained significantly more IL-1beta than any of the other treatment groups, with the level found in these animals being 3-fold higher than normal and 2-fold higher than -VDP. Extracts were also tested in the bioassay to determine the level of active cytokine present. All growth plate extracts contained activity which altered GAG and proteinase release by lapine articular cartilage. Extracts from -VDP-, -VDP+1,25-, and -VDP+Ergo-treated rats stimulated a 40% increase in glycosaminoglycan release compared with extracts from normal rats. In contrast, extracts from -VDP+24,25-treated rats stimulated a 300% increase in glycosaminoglycan release. Both collagenase and neutral metalloproteinase activity of lapine cartilage were increased after incubation with the growth plate extracts. Collagenase activity was significantly increased 8- to 13-fold by the addition of extracts from -VDP-, -VDP+24,25-, or -VDP+1,25 treated animals. Neutral metalloproteinase activity was similarly increased by 4- to 10-fold. To characterize this activity further, growth plate extracts were incubated with neutralizing antibody to IL-1alpha or beta prior to addition to the lapine articular cartilage cultures. When antibodies were used separately, only partial inhibition was observed; incubation with both antibodies blocked 25% of the glycosaminoglycan release observed without antibody and greater than 80% of the enzyme activity released by the articular cartilage cultures. The results of this study show that growth plate cartilage contains both IL-1alpha and beta and indicate that vitamin D regulates the level of IL-1 in this tissue. PMID- 9333118 TI - Enhanced nonsaturable calcium transport in the jejunum of rats during lactation, but not during pregnancy. AB - The lactating (L) rat loses in excess of 100 mg of calcium (Ca) per day to milk at peak lactation. Most of the Ca must be provided by increased intestinal absorption. In an effort to examine adaptation of intestinal calcium absorption during lactation, nonsaturable absorption from the small intestine of rats was calculated from the disappearance of Ca from in situ ligated loops of jejunum during the last week of pregnancy and throughout lactation and weaning. Efficiency of absorption is reflected by the slope of the regression line of Ca absorbed on Ca introduced into the loop. Absorption of Ca in the jejunum was markedly enhanced starting at 5 days of lactation and for the remainder of lactation. Two days after weaning, the efficiency ofjejunal Ca absorption decreased to the nonmated (NM) control level, while the lactation-associated intestinal hypertrophy persisted beyond 2 days postweaning. The percentages of water and Ca absorbed were positively and significantly correlated in both L and NM rats. In contrast to Ca, magnesium (Mg) and strontium (Sr) transport from ligated loops were not enhanced during lactation. Fifty millimolar glucose in the test solution increased the absorption of both water and Ca, but not Mg, from jejunal loops of NM rats. Glucose increased Ca absorption in NM rats up to the level seen in L rats. Glucose did not increase Ca absorption further in L rats, perhaps because of the greater availability of glucose to the intestine during lactation. We conclude that in rats the efficiency of nonsaturable Ca absorption from the jejunum is significantly increased during well established lactation, but not during pregnancy. The underlying mechanism appears to be specific for Ca, may be dependent on glucose, and is unrelated to intestinal hypertrophy. PMID- 9333117 TI - Analysis of osteocalcin expression in transgenic mice reveals a species difference in vitamin D regulation of mouse and human osteocalcin genes. AB - A line of transgenic mice expressing a human osteocalcin genomic fragment (hOClocus) and a murine MC3T3-E1 cell line containing a stably integrated human osteocalcin promoter construct have been developed to characterize the osteogenic and hormonal regulation of human osteocalcin in vivo and in vitro. In this study, we used these models to demonstrate a species difference in the regulation of the mouse and human osteocalcin genes by vitamin D. Repeated administration of 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) to mice carrying the hOClocus transgene resulted in striking increases in serum human osteocalcin, whereas serum mouse osteocalcin levels were unchanged after 24 h and only modestly increased 48 h after the second dose of hormone. 1,25(OH)2D3 increased human calvarial mRNA expression by 1.8-fold and slightly decreased mouse osteocalcin mRNA levels by approximately 1.2-fold. Furthermore, treatment of primary calvarial osteoblasts from these mice with 1,25(OH)2D3 increased human osteocalcin production but inhibited mouse osteocalcin protein accumulation. To investigate further the mechanism for the apparent species difference in vitamin D3 induction of mouse and human osteocalcin, we examined the effect of 1,25(OH)2D3 in an MC3T3-E1 cell line (MC4) containing a stably integrated 3900 bp osteocalcin promoter-luciferase construct. Treatment of MC4 cells with ascorbic acid resulted in parallel increases of the endogenous mouse osteocalcin protein and luciferase reporter activity over a 12-day period. Continuous exposure of MC4 cells to 1,25(OH)2D3 resulted in time-and dose-dependent increases in the activity of the phOC3900 luciferase construct. By contrast, the hormone had no effect on mouse osteocalcin protein concentrations and inhibited its induction by ascorbic acid. However, when cells were treated acutely with 1,25(OH)2D3 at later times during growth in ascorbic acid, the induction of mouse osteocalcin protein was only partially inhibited. In conclusion, our results indicate that common osteogenic signals regulate both mouse and human osteocalcin gene expression, but the mouse gene is resistant to induction by vitamin D. This species difference in vitamin D regulation of osteocalcin appears to result from the failure of 1,25(OH)2D3 to transcriptionally activate the mouse osteocalcin gene. PMID- 9333119 TI - Recombinant transforming growth factor-beta1 induces endochondral bone in the baboon and synergizes with recombinant osteogenic protein-1 (bone morphogenetic protein-7) to initiate rapid bone formation. AB - Several members of the bone morphogenetic protein (BMP) and transforming growth factor-beta (TGF-beta) families are molecular regulators of cartilage and bone regeneration, although their actual roles and combined interactions in skeletal repair are poorly understood. The presence of several molecular forms suggests multiple functions in vivo as well as synergistic interactions during both embryonic bone development and regeneration of cartilage and bone in postfetal life. Here we show for the first time that recombinant human transforming growth factor-beta1 (TGF-beta1) induces endochondral bone formation in extraskeletal sites of adult baboons. We also show that TGF-beta1 and recombinant human osteogenic protein-1 (OP-1, bone morphogenetic protein-7) synergize in inducing large ossicles in extraskeletal sites of the primate as early as 15 days after implantation. A single application of OP-1, in conjunction with an insoluble collagenous matrix as carrier (5, 25, and 125 microg/100 mg of carrier matrix) induced bone differentiation in the rectus abdominis of the baboon. This level of tissue induction was raised several-fold by the simultaneous addition of comparatively low doses of TGF-beta1 (0.5, 1.5, and 5 microg), which by itself induces bone formation in the rectus abdominis at doses of 5 microg/100 mg of carrier matrix. Combinations of OP-1 and TGF-beta1 yielded a 2- to 3-fold increase in cross-sectional area of the newly generated ossicles, with markedly elevated key parameters of bone formation, and corticalization of the newly formed bone by day 15, culminating in bone marrow generation by day 30. The tissue generated by the combined application of OP-1 and TGF-beta1 showed distinct morphological differences when compared with OP-1-treated specimens, with large zones of endochondral development and extensive bone marrow formation. At the doses tested, synergy was optimal at a ratio of 1:20 by weight of TGF beta1 and OP-1, respectively. These results provide evidence for a novel function of TGF-beta1 in the primate and the scientific basis for synergistic molecular therapeutics for the rapid regeneration of cartilage and bone. PMID- 9333120 TI - The mitogenic effect of parathyroid hormone is associated with E2F-dependent activation of cyclin-dependent kinase 1 (cdc2) in osteoblast precursors. AB - Injections of parathyroid hormone (PTH) have been reported to stimulate skeletal accretion through increased bone formation in several species, and osteoblast proliferation is a critical component of bone formation. However, the biological mechanisms of PTH-stimulated bone cell proliferation are largely unknown. In this study, we demonstrated that PTH stimulates proliferation of the osteoblast precursor cell line, TE-85, in association with increasing cdc2 protein levels and its kinase activity. cdc2 antisense oligonucleotides blocked PTH-induced DNA synthesis and cell cycle progression. Analysis of the time course of PTH stimulated cdc2 message levels demonstrated that cdc2 mRNA levels were increased 1.5- to 4-fold between 3-18 h following release from cell synchronization. Transfections of TE-85 cells with a series of cdc2 promoter-luciferase deletion constructs revealed PTH stimulation of the cdc2 promoter. Promoter constructs containing a mutation in the E2F binding site were not stimulated by PTH. Gel mobility shift assays demonstrated increased free E2F levels in TE-85 nuclear extracts in response to PTH. Furthermore, the ratios of hyperphosphorylated to hypophosphorylated forms of Rb protein were increased by PTH treatment. These results demonstrate that PTH-stimulated cdc2 expression was associated with TE-85 cell proliferation and that the mechanism of stimulating cdc2 gene expression involved increasing the levels of free E2F. PMID- 9333121 TI - Stimulatory effects of basic fibroblast growth factor and bone morphogenetic protein-2 on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. AB - Bone marrow stroma contains multipotential mesenchymal progenitor cells which can differentiate into osteoblastic cells; we refer to these cells as mesenchymal stem cells (MSCs). Basic fibroblast growth factor (bFGF) and bone morphogenetic protein-2 (BMP-2) have been implicated in the osteogenic regulatory process by virtue of their mitogenic and differentiation activities, respectively. This study examines and compares the effects of bFGF and BMP-2 on dexamethasone (Dex) dependent in vitro osteogenic differentiation of rat marrow-derived MSCs. A 6-day exposure to bFGF markedly stimulated cell growth and induced osteoblastic differentiation as shown by osteocalcin mRNA expression (day 14), bone nodule formation (day 18), and calcium deposition (day 18). These results indicate that bFGF enhances both mitogenic activity and osteogenic development of Dex-treated marrow MSCs. In contrast, BMP-2 did not induce osteogenesis as strongly as bFGF. Thus, exposure to BMP-2 slightly increased bone nodule number and calcium content compared with the control. Exposure of MSCs to both BMP-2 and bFGF induced expression of osteocalcin mRNA and mineralizing bone-like nodules as early as day 11 and resulted in enhancement of bone formation more markedly than either factor alone. Consistent with these results, porous calcium phosphate ceramic cubes implanted in vivo, which were loaded with MSCs pre-exposed to both bFGF and BMP 2, showed higher histologic score for bone formation than those with MSCs pre exposed to either bFGF or BMP-2 alone. These data indicate that combined treatment with bFGF and BMP-2 synergistically enhances the osteogenic potency of bFGF in rat marrow MSC culture. PMID- 9333122 TI - [3H]bradykinin receptor-binding, receptor-recycling, and receptor-internalization of the B2 bradykinin receptor in the murine osteoblast-like cell line MC3T3-E1. AB - Bradykinin (BK) has been demonstrated to induce inositol phosphate production, release of intracellular Ca2+, and prostaglandin E2 (PGE2) synthesis in the murine osteoblast-like cell line MC3T3-E1. Because cellular response to BK is a function of receptor affinity, receptor coupling, and receptor recycling, we investigated kinetic properties, specificity, and regulation at the BK-receptor level on intact, BK-sensitive MC3T3-E1 cells. Our results clearly demonstrate the existence of a single category of binding sites for [3H]BK (kD =366+/-98 pM; Bmax =45.3+/-6.6 fmol/mg of protein). Displacement studies with various BK analogs gave a rank order compatible with a B2 BK-receptor type (BK > Lys-BK > [Hyp3]-BK > Met-Lys-BK > HOE140 > Tyr-BK > Tyr8-BK > D-Arg, [Hyp3, Thi5,8, D-Phe7]-BK > [D Phe7]-BK > des-Arg9-BK > des-Arg9, [Leu8]-BK = angiotensin II). No atypic high affinity binding sites for the B1 receptor agonist des-Arg9-BK could be observed. Prestimulation of MC3T3-E1 cells with BK resulted in the disappearance of accessible B2 receptors at the cell surface by internalization. Postexposure of BK-pretreated cells to ligand-free medium resulted in almost complete receptor restoration within 30 minutes, exhibiting an intermediate state of two categories of binding sites (kD1 =444+/-37 pM, Bmax1 =9.2+/-0.3 fmol/mg of protein and kD2 =2.7+/-0.28 pM, Bmax2 =24.2+/-0.2 fmol/mg of protein), probably representing coupled and uncoupled B2 receptors. Prolonged stimulation with BK (2.5-5 h) also revealed the temporal occurrence of two categories of binding sites after 2.5 h (kD1 =228+/-3.5 pM; Bmax1 =15.6+/-0.6 fmol/mg of protein; kD2 =2.7+/-0.25 nM; Bmax2 =40.7+/-1.5 fmol/mg of protein), whereas low-affinity binding sites disappeared after 5 h. PMID- 9333123 TI - Signal transduction of mechanical stimuli is dependent on microfilament integrity: identification of osteopontin as a mechanically induced gene in osteoblasts. AB - Mechanical perturbation has been shown to modulate a wide variety of changes in second message signals and patterns of gene expression in osteoblasts. Embryonic chick osteoblasts were subjected to a dynamic spatially uniform biaxial strain (1.3% applied strain) at 0.25 Hz for a single 2-h period, and osteopontin (OPN), an Arg-Gly-Asp (RGD)-containing protein, was shown to be a mechanoresponsive gene. Expression of opn mRNA reached a maximal 4-fold increase 9 h after the end of the mechanical perturbation that was not inhibited by cycloheximide, thus demonstrating that mechanoinduction of opn expression is a primary response through the activation of pre-existing transcriptional factors. The signal transduction pathways, which mediated the increased expression of opn in response to mechanical stimuli, were shown to be dependent on the activation of a tyrosine kinase(s) and protein kinase A (PKA) or a PKA-like kinase. Selective inhibition of protein kinase C (PKC) had no effect on the mechanoinduction of osteopontin even though opn has been demonstrated to be an early response gene to phorbol 12 myristate 13-acetate (PMA) stimulation. Mechanotransduction was dependent on microfilament integrity since cytochalasin-D blocked the up-regulation of the opn expression; however, microfilament disruption had no effect on the PMA induction of the gene. The microtubule component of the cytoskeleton was not related to the mechanism of signal transduction involved in controlling opn expression in response to mechanical stimulation since colchicine did not block opn expression. Mechanical stimulus was shown to activate focal adhesion kinase (FAK), which specifically became associated with the cytoskeleton after mechanical perturbation, and its association with the cytoskeleton was dependent on tyrosine kinase activity. In conclusion, these results demonstrate that the signal transduction pathway for mechanical activation of opn is uniquely dependent on the structural integrity of the microfilament component of the cytoskeleton. In contrast, the PKC pathway, which also activates this gene in osteoblasts, acts independently of the cytoskeleton in the transduction of its activity. PMID- 9333124 TI - Fas and Fas ligand interaction is necessary for human osteoblast apoptosis. AB - We investigated the cellular and humoral interactions between peripheral blood mononuclear cells (PBMCs) and human osteoblasts, leading to apoptosis of osteoblasts. Human osteoblastic cell line MG63 and human primary osteoblast-like cells obtained from biopsy specimens were used in this study. PBMCs were isolated from healthy donors and cultured with or without stimulation by recombinant interleukin-2 followed by 12-o-tetradecanoylphorbol 13-acetate with ionomycin. Fas was functionally expressed on MG63 and primary osteoblast-like cells. Activated PBMCs expressed Fas ligand (FasL) strongly on their surface and killed MG63 and primary osteoblast-like cells. Cultured supernatants of activated PBMCs also induced apoptotic cell death of MG63 and primary osteoblast-like cells. In contrast, both unstimulated PBMCs and cultured supernatants of unstimulated PBMCs did not induce apoptosis of these cells. Furthermore, the cytotoxic effect and induction of apoptosis against MG63 and primary osteoblast-like cells by activated PBMCs and cultured supernatants were inhibited significantly by human Fas chimeric protein. Our data showed that human osteoblasts expressed Fas fuctionally and both membrane-type and soluble form FasL from activated PBMCs induced apoptosis of these cells, providing the one possible mechanism of bone loss in inflammatory diseases such as rheumatoid arthritis. PMID- 9333125 TI - Further observations on programmed cell death in the epiphyseal growth plate: comparison of normal and dyschondroplastic epiphyses. AB - The objective of the investigation was to provide information on apoptosis in the normal epiphysis and to assess apoptosis in the plate of the dyschondroplastic chick. Apoptosis was evaluated using two terminal deoxynucleotide transferase end labeling procedures, DNA fragmentation and nuclear morphology. We found that there was a minimal level of apoptosis in the dyschondroplastic cartilage. In the tibial dyschondroplastic (TD) lesion itself, only about 3% of cells are positive in the articular and proliferative regions; 11% of prehypertrophic chondrocytes are stained by the end-labeling procedure, and most of the cells are localized around vascular channels at the calcifying front. This finding suggests that dyschondroplasia is linked to impairment of apoptosis, and as a result the tissue contains immature cells that have outlived their normal life span. In contrast, in the normal plate, we noted that when the proliferative period was complete, the cells became terminal transferase positive; in addition, chondrocytes in the normal plate exhibited DNA fragmentation. Semiquantitative analysis of stained chondrocytes in the growth plate indicate that in the proliferative zone 15.5% of cells are terminal deoxynucleotidyl transferase (TUNEL) positive; in contrast, 44% of postmitotic chondrocytes are stained by the TUNEL procedure. The presence of a sharp border between the pre- and postmitotic zones suggests that the stimulus for apoptosis is maturation dependent and reflects local metabolic control. We also examined apoptosis in metaphyseal osteoblasts. We found that adjacent to the epiphysis, many osteoblasts were undergoing apoptosis. In more mature sites in the metaphysis, there was less cell death, indicating that osteoblast apoptosis was delayed and cells were completing their normal life cycle. Although terminal transferase end-labeled cells were not seen in articular cartilage, we noted that fibroblasts, in the perichondrial ligament surrounding the articular as well as the epiphyseal regions of the plate, were undergoing apoptosis. Apoptosis at this site may be related to lateral expansion of the cartilages, reflect a high cell turnover rate at the junction between the tissues, and result from paracrine signals received from the underlying cartilage. PMID- 9333126 TI - Chondrocytes are regulated by cellular adhesion through CD44 and hyaluronic acid pathway. AB - The articular cartilage consists of resident chondrocytes embedded within the extracellular matrix which contains several components such as collagen and hyaluronic acids (HA). CD44 is a major cell surface receptor for HA and is homologous to cartilage-link proteins. Although CD44 is present in cartilage, it is not clear if chondrocytes adhere to HA through CD44 or whether such adhesion changes the function of chondrocytes. We studied the molecular mechanisms of CD44 related chondrocyte adhesion to HA and the effects of such adhesion on chondrocyte function. Experiments were performed using the human chondrosarcoma derived chondrocyte-like cell line HCS-2/8. Our results showed that (a) HCS-2/8 cells highly expressed CD44; (b) HCS-2/8 cells efficiently adhered to HA without any stimuli; (c) monoclonal antibody (mAb)-blocking studies indicated that adhesion of HCS-2/8 cells to HA was mainly mediated by the CD44/HA pathway; (d) cellular adhesion to HA increased the proliferation of HCS-2/8 cells, independent of transforming growth factor-beta (TGF-beta), but this was inhibited by CD44 mAb; (e) the adhesion of chondrocytes to HA also induced c-myc mRNA expression and this was also inhibited by CD44 mAb; and (f) the adhesion of cells to HA augmented TGF-beta mRNA expression, a process also reduced by CD44 mAb. Thus, HCS 2/8 cells effectively adhered to HA through cell surface CD44. The adhesion was also involved in cellular signaling which induced cellular proliferation and expression of c-myc mRNA as well as TGF-beta mRNA expression within the cells. Our results indicate that CD44 on chondrocytes plays an important role in normal and abnormal functions of cartilage through its adhesion to HA, which induces a variety of stimulatory signals to regulate chondrocyte proliferation as well as matrix synthesis in cartilage microenvironment. PMID- 9333127 TI - Effect of metabolic acidosis on the potassium content of bone. AB - Metabolic acidosis induces resorption of cultured bone, resulting in a net efflux of calcium (Ca) from the bone and an apparent loss of mineral potassium (K). However, in these organ cultures, there is diffusion of K between the medium and the crystal lattice, causing difficulty in interpretation of the acid-induced changes in mineral ion composition. To determine the effects of acidosis on bone mineral K, we injected 4-day-old neonatal mice with pure stable isotope 41K, equal to approximately 5% of their total body K. Calvariae were dissected 24 h later and then cultured for 24 h in medium without added 41K, either at pH approximately 7.4 (Ctl) or at pH approximately 7.1 (Ac), with or without the osteoclastic inhibitor calcitonin (3 x 10(-9) M, CT). The bone isotopic ion content was determined with a high-resolution scanning ion microprobe utilizing secondary ion mass spectrometry. 41K is present in nature at 6.7% of total K. The injected 41K raised the ratio of bone 41K/(39K+41K) to 9.8+/-0.5% on the surface (ratios of counts per second of detected secondary ions, mean+/-95% confidence interval) but did not alter the ratio in the interior (6.9+/-0.4%), indicating biological incorporation of the 41K into the mineral surface. The ratios of 41K/40Ca on the surface of Ctl calvariae was 14.4+/-1.2, indicating that bone mineral surface is rich in K compared with Ca. Compared with Ctl, Ac caused a marked increase in the net Ca efflux from bone that was blocked by CT. Ac also induced a marked fall in the ratio of 41K/40Ca on the surface of the calvariae (43+/-0.5, p < 0.01 vs. Ctl), which was partially blocked by CT (8.2+/-0.9, p < 0.01 vs. Ctl and vs. Ac), indicating that Ac causes a greater release of bone mineral K than Ca which is partially blocked by CT. Thus, bone mineral surface is rich in K relative to Ca, acidosis induces a greater release of surface mineral K than Ca, and osteoclastic function is necessary to support the enriched levels of surface mineral K in the presence of acidosis. PMID- 9333128 TI - Growth hormone normalizes renal 1,25-dihydroxyvitamin D3-24-hydroxylase gene expression but not Na+-phosphate cotransporter (Npt2) mRNA in phosphate-deprived Hyp mice. AB - The murine X-linked Hyp mutation is characterized by decreased renal expression of type II Na+-phosphate (Pi) cotransporter (Npt2) mRNA and an abnormal vitamin D response to Pi deprivation. The latter is manifest by an aberrant fall in serum 1,25-dihydroxyvitamin D3 (1,25(OH)2D) levels that is associated with an increase in renal 1,25(OH)2D-24-hydroxylase (24-hydroxylase), the first enzyme in the C-24 oxidation pathway. Because growth hormone (GH) enhances renal Na+-Pi cotransport and permits the adaptive 1,25(OH)2D response in Pi-deprived hypophysectomized rats, we examined the effects of GH on vitamin D metabolism and renal Npt2 mRNA abundance in Hyp mice fed control and low Pi diets. GH significantly decreased renal 24-hydroxylase activity (0.202+/-0.020 to 0.098+/-0.008 pmol/mg of protein/minute, p < 0.05) and mRNA abundance, relative to beta-actin mRNA (299+/ 13 to 78+/-14, p < 0.05), in Hyp mice fed the low Pi diet but had no effect on either parameter in mutants fed the control diet. Moreover, after GH treatment, renal 24-hydroxylase gene expression was no longer elevated in Pi-deprived Hyp mice relative to mutants fed control diet. In contrast, GH did not correct the serum concentration of 1,25(OH)2D in Pi-deprived Hyp mice. We also demonstrate that GH did not normalize renal Npt2 mRNA expression, relative to beta-actin mRNA, in Hyp mice fed either control or low Pi diets. The present data demonstrate that normalization of renal 24-hydroxylase gene expression in Pi deprived Hyp mice by GH is not sufficient to correct the serum concentration of 1,25(OH)2D and is not associated with an alteration in renal Npt2 mRNA expression. PMID- 9333129 TI - Phosphorylation of the human calcitonin receptor by multiple kinases is localized to the C-terminus. AB - The calcitonin receptor is a seven-transmembrane G-protein coupled receptor which is located on osteoclasts, in kidney, and in brain. The receptor signals through multiple pathways, including activation of adenylate cyclase, leading to inhibition of bone resorption. In the present study, we used antibodies raised against the C-terminus of the human calcitonin (CT) receptor to study receptor phosphorylation. In baby hamster kidney cells transfected with the human CT receptor, phosphorylation of the receptor increased approximately 2.5-fold after cells were treated with calcitonin, phorbol ester, forskolin, or calcitonin plus phorbol ester. Phosphorylation reached a maximum 20 minutes after treatment with sCT and half-maximal phosphorylation was observed at 0.1 nM sCT, a hormone concentration related to receptor occupancy. Digestion of the immunoprecipitated receptor with cyanogen bromide (CNBr) yielded a single 32P-labeled fragment which migrates at Mr 14 kD on gel electrophoresis. This corresponds to the predicted size of the CNBr fragment containing the C-terminal domain of the receptor. No 32P-labeled bands were observed for CNBr fragments predicted to contain the first, second, or third intracellular loops. An identical labeling pattern was seen with cells expressing an alternatively spliced isoform of the human receptor (insert-positive isoform). Phosphorylation of the receptor by phorbol ester and forskolin was further localized to a Mr 6 kD proteolytic fragment within the C terminus. The protein kinase A and C inhibitors staurosporine, chelerythrine, and H-89 had little effect on CT-induced phosphorylation, suggesting that nonsecond messenger-activated kinases are involved in hormone-dependent CT receptor phosphorylation. PMID- 9333130 TI - A collagenous cementum-derived attachment protein is a marker for progenitors of the mineralized tissue-forming cell lineage of the periodontal ligament. AB - The periodontal ligament (PDL) is a fibrous and cellular connective tissue that mediates tooth attachment to bone, and it comprises fibroblastic and mineralized tissue-forming (MTF) progenitors. The MTF progenitors are believed to give rise to the cementoblastic and osteoblastic lineages. Cementum attachment protein (CAP) is a collagenous cementum-derived protein which binds strongly to osteoblasts, moderately to PDL cells, and weakly to gingival fibroblasts. The aim of the present study was to determine the relationship between the capacity of PDL progenitors to bind CAP and their potential to express alkaline phosphatase (ALP) and form mineralized-like tissue in culture. Cloned human PDL progenitor populations obtained from nine human donors were assayed for their constitutive capacity to bind CAP and express ALP, and for the dexamethasone-induced potential to form mineralized-like tissue in culture in the presence of ascorbic acid and beta-glycerophosphate. Forty percent of the progenitor clones produced mineralized-like tissue. Two patterns of mineralization were observed: a spread and flat pattern similar to that produced by human bone cells in culture and a nodular ridge-like type resembling that formed by human cementoma-derived cells. A direct correlation was found between the percentage of ALP positive cells in each progenitor clone and the amount of mineralized-like tissue formed (r = 0.565). Similar correlations were found between the number of ALP positive cells and the binding capacity of each clone (r = 0.392) and between the CAP binding capacity and mineralized-like tissue formation (r = 0.584). Multiple regression analysis indicated that the constitutive capacity of a clone to bind CAP and express ALP is directly correlated to its dexamethasone-induced potential to form mineralized tissue (r = 0.675). These results indicate that CAP binding and ALP expression can serve as markers for the identification of MTF progenitors in the heterogeneous cultured population of the human periodontal ligament. These data show for the first time that binding capacity to extracellular components of mineralized tissues can be a marker for mineralized tissue-forming progenitors. PMID- 9333131 TI - Elimination and biochemical responses to intravenous alendronate in postmenopausal osteoporosis. AB - Postmenopausal women with established vertebral osteoporosis were studied for 2 years to determine the terminal elimination half-life and the duration of response to treatment with intravenous alendronate (30 mg) given over 4 days. The urinary excretion of alendronate followed a multiexponential decline. Approximately 50% of the total dose was excreted over the first 5 days, and a further 17% was excreted in the succeeding 6 months. Thereafter, there was a much slower elimination phase with an estimated mean terminal half-life of greater than 10 years (n = 11). Urinary excretion of hydroxyproline and calcium decreased significantly from pretreatment values by day 3, reaching a nadir by 1 week (40% and 67% decrease, respectively). Thereafter, hydroxyproline remained suppressed for the following 2 years. In contrast, urinary calcium excretion returned gradually toward pretreatment values over the first year and during the second year was comparable to pretreatment values. Serum activity of alkaline phosphatase activity decreased over 3 months (23% reduction), increased gradually thereafter, and returned to pretreatment values at month 24. Bone mineral density measured at the spine increased by approximately 5% during the first year and remained significantly higher than pretreatment values at 2 years. We conclude that a short course of high doses of intravenous alendronate is associated with a prolonged skeletal retention of the agent. This open study also suggests that this regimen has a sustained effect on bone turnover persisting for at least 1 year. PMID- 9333132 TI - Evidence for increased bone formation following a brief endurance-type training intervention in adolescent males. AB - The effect of exercise training, particularly relatively brief periods, on bone turnover markers in adolescents has been poorly studied. Thirty-eight healthy males (16+/-0.7 years) participated in a 5-week summer school program in which 20 subjects were randomly assigned to a training group consisting of 2 h/day, 5 days/week of endurance exercise, and 18 subjects were assigned to a control group. Bone formation was assessed by measurements of circulating osteocalcin, bone-specific alkaline phosphatase (BSAP), and the C-terminal procollagen peptide (PICP). Bone resorption was assessed by urinary levels of free deoxypyridinoline cross-links (dPYR) and the C-(CTX) and N-terminal (NTX) telopeptide cross-links. Prior to training, there was a weak positive correlation between fitness and PICP (r = 0.27, p < 0.05), but no correlations were observed between fitness and either the other markers of bone formation or bone resorption. Training led to a significant increase in (1) osteocalcin (15+/-4%, p < 0.03), (2) BSAP (21+/-6%, p < 0.02), and (3) PICP (30+/-11%, p < 0.03) and to a significant decrease in NTX ( 21 +/- 3%, p < 0.05). These bone turnover markers did not change in the control subjects (osteocalcin, 0+/-4%; BSAP, 2+/-4%; PICP, -4 +/- 6%; NTX, -6 +/- 4%). There was no change in urinary dPYR and CTX in either control or trained subjects. Fitness is only weakly, if at all, correlated with bone formation, but a relatively brief period of endurance training leads to a substantial increase in bone formation markers in adolescent males. School-based, short-term exercise training programs could play a role in enhancing bone formation in adolescents. PMID- 9333133 TI - Quantification of biochemical markers of bone turnover by kinetic measures of bone formation and resorption in young healthy females. AB - The quantification of biochemical markers of bone formation and resorption with kinetic measures of bone turnover is an essential step in their validation. Some biochemical markers have been validated by quantification against formation and resorption rates measured by calcium kinetics in adults with bone disease. However, none has been validated in healthy individuals who are undergoing skeletal growth and bone consolidation. Therefore, we have measured biochemical markers of bone formation (serum osteocalcin [OC], bone-specific alkaline phosphatase [BAP], and total alkaline phosphatase [ALP]) and resorption (serum tartrate resistant acid phosphatase [TRAP], urinary cross-linked N teleopeptides of type I collagen/creatinine [NTx/Cr], and hydroxyproline/creatinine [OHP/Cr]) in healthy females aged 11-32 years (n = 31) after an overnight fast to determine their relationship with bone formation (Vo+) and bone resorption (Vo-) as measured by calcium kinetics and balance. All biochemical markers were highly intercorrelated (r > 0.6, p < 0.001) as were Vo+ and Vo- (r = 0.91, p < 0.001). Highly significant correlations were present between bone formation measured by calcium kinetics (Vo+) and serum levels of bone biochemical markers (OC, r = 0.82, p = 0.001; ALP, r = 0.92, p = 0.001; and BAP, r = 0.90, p = 0.001) and between bone resorption measured by calcium kinetics (Vo-) and fasting serum levels and urine creatinine ratios of biochemical markers (TRAP, r = 0.77, p < 0.001; OHP/Cr, r = 0.79, p < 0.001; and NTx/Cr, r = 0.70, p < 0.001). Thus, biochemical markers of bone formation and resorption can be used to predict calcium kinetic rates during skeletal growth and the early years of formation of peak bone mass, ages at which strategies to build peak bone mass are important. Biochemical markers of formation and resorption are equally useful in predicting either the bone formation rate or the resorption rate. PMID- 9333134 TI - Prediction of vertebral strength in vitro by spinal bone densitometry and calcaneal ultrasound. AB - Spinal bone mineral density (BMD) measurements and calcaneal ultrasound were compared in terms of their ability to predict the strength of the third lumbar vertebral body using specimens from 62 adult cadavers (28 females, 34 males). BMD was measured using dual X-ray absorptiometry (DXA) in both vertebra and calcaneus. Quantitative computed tomography (QCT) was used to determine trabecular BMD, cortical BMD, cortical area, and total cross-sectional area (CSA) of the vertebral body. Bone velocity (BV) and broadband ultrasonic attenuation (BUA) were measured in the right calcaneus. Vertebral strength was determined by uniaxial compressive testing. Vertebral ultimate load was best correlated with DXA-determined vertebral BMD (r2 = 0.64). Of the QCT parameters, the best correlation with strength was obtained using the product of trabecular BMD and CSA (r2 = 0.61). For vertebral ultimate stress, however, the best correlation was observed with QCT-measured trabecular BMD (r2 = 0.51); the correlation with DXA determined BMD was slightly poorer (r2 = 0.44). Calcaneal ultrasound correlated only weakly with both ultimate load and stress with correlation coefficients (r2) of 0.10-0.17, as did calcaneal BMD (r2 = 0.18). Both spinal DXA and spinal QCT were significantly (p < 0.001) better predictors of L3 ultimate load and stress than were either calcaneal ultrasound or calcaneal DXA. Multiple regression analysis revealed that calcaneal ultrasound did not significantly improve the predictive ability of either DXA or QCT for L3 ultimate load or stress. Calcaneal DXA BMD, bone velocity, and BUA correlated well with each other (r2 = 0.67-0.76), but were only modestly correlated with the DXA and QCT measurements of the vertebra. These data indicate that spinal DXA and spinal QCT provide comparable prediction of vertebral strength, but that a substantial proportion (typically 40%) of the variability in vertebral strength is unaccounted for by BMD measurements. Ultrasonic measurements at the calcaneus are poor predictors of vertebral strength in vitro, and ultrasound does not add predictive information independently of BMD. These findings contrast with emerging clinical data, suggesting that calcaneal ultrasound may be a valuable predictor of vertebral fracture risk in vivo. A possible explanation for this apparent discrepancy between in vivo and in vitro findings could be that current clinical ultrasound measurements at the calcaneus reflect factors that are related to fracture risk but not associated with bone fragility. PMID- 9333135 TI - Skeletal involvement in female acromegalic subjects: the effects of growth hormone excess in amenorrheal and menstruating patients. AB - Bone involvement is a common clinical feature in acromegalic patients, though previous studies gave divergent results possibly because of the different gonadal status of the patients studied. To study the influence of estrogen milieu in these patients, we evaluated 23 acromegalic patients with active disease, subdivided into two groups: menstruating and amenorrheal patients, comparable for duration and activity of disease. Forty-two matched women served as controls. Skeletal involvement was studied by measuring: (a) the main biomarkers of bone turnover: serum alkaline phosphatase total activity (AP), bone GLA protein (BGP), serum carboxy-terminal propeptide of type I collagen (PICP), serum type I cross linked N-telopeptide (ICTP), and urinary pyridinoline and deoxypyridinoline corrected for creatinine (Pyr/Cr, D-Pyr/Cr) and urinary calcium/creatinine ratio (Ca/Cr); (b) bone mineral density (BMD), as measured by quantitative computed tomography both at lumbar spine and distal radius, and by dual X-ray absorptiometry both at lumbar spine and at three femoral sites (Ward's triangle, femoral neck, and great trochanter). AP, BGP, ICTP, Pyr/Cr, D-Pyr/Cr were significantly higher in patients than in controls, independent of the menstrual pattern. Higher PICP levels were found in the whole group and in menstruating acromegalics when compared with control women; no difference was found in amenorrheal patients, who in turn showed higher urinary Ca/Cr values. When patients were considered all together, BMD at spine, femoral neck, and trochanter was higher than in controls. In contrast, when the gonadal status was taking into account and, menstruating and amenorrheal subjects were considered separately, BMD at spine, but not in other sites, was significantly higher in menstruating patients than in controls. In contrast, no difference of BMD values at any site was observed between amenorrheal patients and controls. The mean BMD Z scores allowed us to detect an unequal involvement of different skeletal sites. Our results show that bone turnover is increased in acromegalic women and suggest that GH anabolic effect on bone is more evident in the presence of estrogens and that different skeletal sites may be affected differently by hormone excess. PMID- 9333136 TI - Strain gradients correlate with sites of exercise-induced bone-forming surfaces in the adult skeleton. AB - Physical activity is capable of increasing adult bone mass. The specific osteogenic component of the mechanical stimulus is, however, unknown. Using an exogenous loading model, it was recently reported that circumferential gradients of longitudinal normal strain are strongly associated with the specific sites of periosteal bone formation. Here, we used high-speed running to test this proposed relation in an exercise model of bone adaptation. The strain environment generated during running in a mid-diaphyseal tarsometatarsal section was determined from triple-rosette strain gages in six adult roosters (>1 year). A second group of roosters was run at a high speed (1500 loading cycles/day) on a treadmill for 3 weeks. Periosteal surfaces were activated in five out of eight animals. Mechanical parameters as well as periosteal activation (as measured by incorporated fluorescent labels) were quantified site-specifically in 12 30 degrees sectors subdividing a mid-diaphyseal section. The amount of periosteal mineralizing surface per sector correlated strongly (R2 = 0.63) with the induced peak circumferential strain gradients. Conversely, peak strain magnitude and peak strain rate were only weakly associated with the sites of periosteal activation. The unique feature of this study is that a specific mechanical stimulus (peak circumferential strain gradients) was successfully correlated with specific sites of periosteal bone activation induced in a noninvasive bone adaptation model. The knowledge of this mechanical parameter may help to design exercise regimens that are able to deposit bone at sites where increased structural strength is most needed. PMID- 9333137 TI - Long-term effects of intravenous pamidronate in fibrous dysplasia of bone. AB - Fibrous dysplasia of bone (FD) is a rare disorder characterized by proliferation of fibrous tissue in bone marrow leading to osteolytic lesions. It causes bone pain and fractures. To date the only treatment is orthopedic. Histological and biochemical similarities between FD and Paget's bone disease related to increased osteoclastic resorption led us to propose treatment with the bisphosphonate pamidronate. The aim of the study was to assess the long-term effects of intravenous pamidronate in FD. In this open label phase III study, 20 patients with FD (11 males and 9 females; mean age 31 years) received courses of 180 mg of intravenous pamidronate every 6 months (60 mg/day during 3 days by infusion). The mean duration of follow-up was 39 months (range 18-64). Severity of bone pain, number of painful skeletal sites per patient, X-rays of all involved areas, serum alkaline phosphatase, fasting urinary hydroxyproline, and urinary type I collagen C-telopeptide were assessed every 6 months. The severity of bone pain and the number of painful sites appeared to be significantly reduced. All biochemical markers of bone remodeling were substantially lowered. We observed a radiographic response in nine patients with refilling of osteolytic lesions. A mineralization defect proven by bone biopsy was observed in one case. Four patients sustained bone stress lines, but no fracture occurred. We suggest that intravenous pamidronate alleviates bone pain, reduces the rate of bone turnover assessed by biochemical markers, and improves radiological lesions of FD. Few side effects were observed. PMID- 9333138 TI - The hyperprostaglandin E syndrome: a hypercalciuric variant of Bartter's syndrome. PMID- 9333139 TI - Induction of glutathione-dependent formaldehyde dehydrogenase activity in Escherichia coli and Hemophilus influenza. AB - We have examined the induction of glutathione-dependent formaldehyde dehydrogenase (GS-FDH) activity in Escherichia coli and Hemophilus influenza. Formaldehyde was found to induce enzyme activity in both E. coli and H. influenza at concentrations between 0.6 and 20 ppm. Higher formaldehyde concentrations were toxic. Methanol concentrations up to 20% (200,000 ppm) and sodium formate concentrations up to 2% (20,000 ppm) gave negligible amounts of induction. The basic mechanism of induction was probed by inducing GS-FDH activity in the presence of rifampicin to inhibit RNA synthesis or chloramphenicol to inhibit protein synthesis. Both reagents inhibited GS-FDH induction, demonstrating that regulation occurs at the level of transcription. These results indicate that at least one function of GS-FDH in Gram-negative bacteria is to detoxify exogenous formaldehyde encountered in their environment and that GS-FDH inducibility may be a common feature of Gram-negative bacteria. PMID- 9333140 TI - Spontaneous seroconversion in hepatitis B e antigen-positive chronic hepatitis B: implications for interferon therapy. AB - This study compared rates of spontaneous hepatitis B e antigen (HBeAg)-positive to -negative seroconversion in chronic carriers of hepatitis B virus (HBV) with rates reported during interferon-alpha therapy. Four hundred fifty-four Asian American HBeAg-positive HBV carriers, followed for 1-10 years, were tested approximately every 6 months for HBeAg. Patients with alanine aminotransferase levels > or = 50 IU/mL at entry had 1067.3 seroconversions/10(5) person-months in the 5- to 19-year age group, 1753.3 in the 20- to 34-year group, and 1257.2 in the 35- to 50-year group. Published data indicate that 30% of children and 33% of adults seroconvert during interferon-alpha treatment and follow-up. In our study population, spontaneous seroconversion occurred in 15% of children (95% confidence interval [CI], 8%-27%), 23% of adults 20-34 years (95% CI, 15%-34%), and 17% of adults 35-50 years (95% CI, 10%-28%) during the same interval. The high rate of spontaneous seroconversion should be weighed in decisions to treat HBV carriers with interferon-alpha. PMID- 9333142 TI - Hepatitis C virus-specific cytolytic T lymphocyte and T helper cell responses in seronegative persons. AB - Hepatitis C virus (HCV) is a common infection worldwide, and in most persons, it leads to persistent viremia and liver damage. Efforts to identify the correlates of protective immunity are hampered by this high rate of persistent infection in both infected humans and the only animal model, the chimpanzee. Peripheral blood mononuclear cells from seronegative persons were stimulated with synthetic peptides that represent epitopes recognized by HCV-specific cytotoxic T lymphocytes (CTL) after natural infection. In addition, CD4+ proliferative responses to recombinant HCV proteins were examined in these same persons. CTL responses directed against a peptide epitope of HCV and proliferative responses in 2 HCV-seronegative persons with possible occupational exposure to HCV were found. These otherwise healthy persons were not viremic, suggesting that they may have recovered from acute HCV infection. Characterization of virus-specific immune responses in exposed but seronegative persons may provide important clues as to the nature of protective immunity in HCV. PMID- 9333141 TI - Molecular epidemiology of hepatitis B virus in Central America reflected in the genetic variability of the small S gene. AB - The S genes of 31 Central American hepatitis B virus (HBV) strains belonging to genotypes A, C, D, and F (4, 1, 4, and 22 strains, respectively) were compared with 104 published S genes. According to the deduced S gene product, 21 genotype F strains encoded adw4, while 1 encoded ayw4. Three clusters were revealed within genotype F, which correlated with substitutions at residue 45. In a cluster of 18 Central American and 1 Alaskan strain, all had Thr45. One cluster included 2 Central American strains and 6 strains from South America and Europe, which had Leu45. Two Nicaraguan strains differed by five substitutions, including a Pro45 in the S gene product from other F strains. In conclusion, the dominating HBV genotype was F, which might be the reason for a low prevalence of HBV in the area, despite high prevalence of hepatitis A. These infections otherwise vary in parallel and are considered to reflect socioeconomic conditions. PMID- 9333143 TI - Antigenic structure, function, and evolution of the hemagglutinin-neuraminidase protein of human parainfluenza virus type 1. AB - Twenty-two monoclonal antibodies directed to the hemagglutinin-neuraminidase protein of human parainfluenza virus type 1 (HPIV-1) were used in competition assays to create an antigenic map of neutralization sites. Eighty-seven clinical strains isolated over 35 years from multiple geographic regions were reacted in ELISA, hemagglutinin-inhibition, and microneutralization assays with these monoclonal antibodies. Together these assays revealed 21 epitopes on five nonoverlapping antigenic sites (I, III-VI) with a sixth (II) bridging site connecting sites I, III, and IV. Only 7 (33%) of these epitopes were conserved among all isolates. Previously described HPIV-1 genotypes were associated with the presence or absence of specific antigenic sites and evidence of probable immune selection within genotypes. Two sites were present on all isolates tested (III, V), and one (VI, genotype A) has not been found for 15 years. Forty hemagglutinin-neuraminidase nucleotide sequences were analyzed in terms of homology, structure, and evolution. These data may be useful in future epidemiologic, therapeutic, or vaccine-related work. PMID- 9333144 TI - Seroreactivity to human papillomavirus type 16 virus-like particles is lower in high-risk men than in high-risk women. AB - Seroreactivity to human papillomavirus type 16 (HPV-16) virus-like particles (VLPs) in men attending clinics for sexually transmitted diseases (STDs) in Denmark (n = 219) and Greenland (n = 88) was compared with seroreactivity in women attending the same clinics and was furthermore related to epidemiologic variables and concurrent HPV DNA detection. Risk factors for male seropositivity in Denmark were lifetime number of sex partners, a history of STDs, and sexual preference and in Greenland were ever having had syphilis and years at school. Although men reported significantly more sex partners, the mean seroreactivity was significantly lower in men than in women: 0.50 and 0.75, respectively, in Denmark and 0.53 and 0.86 in Greenland (P = .0001). Male seropositivity was not correlated with concurrent HPV DNA detection, but only 15 Danish and 6 Greenlandic men had HPV-16 DNA. Presence of HPV-16 VLP antibodies appears to be a biomarker for exposure to genital HPVs in men but is less sensitive than in women. PMID- 9333145 TI - Protective immunity against respiratory syncytial virus in early life after murine maternal or neonatal vaccination with the recombinant G fusion protein BBG2Na. AB - Maternal and neonatal immunization were evaluated for their capacity to induce protective immunity against respiratory syncytial virus (RSV) lower respiratory tract infections in early life. Murine models were studied by use of a novel recombinant vaccine candidate, designated BBG2Na, which was derived in part from the RSV (Long) G protein. Maternal immunization resulted in the passive transfer of high levels of RSV-A antibodies to the offspring, which protected them from RSV challenge for up to 14 weeks. Indeed, protection correlated with the detection of RSV antibodies in the serum. Neonatal immunization with BBG2Na induced significant antibody responses even in the first week of life. Most importantly, these neonatal responses were not inhibited by the presence of RSV maternal antibodies. Consequently, the combination of maternal and neonatal immunization with BBG2Na resulted in the continual presence of protective levels of antibodies in the offspring. PMID- 9333146 TI - A randomized, double-blind, placebo-controlled trial of cidofovir gel for the treatment of acyclovir-unresponsive mucocutaneous herpes simplex virus infection in patients with AIDS. AB - The safety and efficacy of cidofovir gel for treatment of acyclovir-unresponsive herpes simplex virus infections in AIDS patients was evaluated in a randomized, double-blind, multicenter trial. Cidofovir (0.3% or 1%) or placebo gel was applied once daily for 5 days. Ten of 20 cidofovir-treated and none of 10 placebo treated patients had complete healing or >50% decreased area (P = .008); 30% of cidofovir-treated patients versus 0 placebo recipients had complete healing (P = .031). Viral shedding ceased in 13 (87%) of 15 cidofovir-treated and 0 of 9 placebo-treated patients (P = .00004). For cidofovir-treated patients, median time to complete or good response was 21 days, and median time to negative viral culture was 2 days (P = .025, P = .0001, respectively). Median lesion area decreases were 58% for cidofovir-treated versus 0 for placebo-treated patients (P = .005), and mean pain score changes were -1.84 versus -0.34 (P = .042). Application site reactions occurred in 25% of cidofovir-treated and 20% of placebo-treated patients; none was dose-limiting. Cidofovir therapy provided significant benefits in lesion healing, virologic effect, and pain reduction. PMID- 9333147 TI - Mutations in the pol gene of human immunodeficiency virus type 1 in infected patients receiving didanosine and hydroxyurea combination therapy. AB - The pattern of mutations in the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) strains that confer resistance to didanosine (ddI) was analyzed in 2 groups of patients receiving either ddI monotherapy or ddI plus hydroxyurea (HU) combination therapy. Twelve patients receiving combination therapy and 8 receiving monotherapy were tested. Combinations of ddI plus HU did not prevent the onset of mutations, which emerged in 50% of the patients in this group compared with 25% of the ddI monotherapy group. In addition, in 1 patient from the combination therapy arm, who had a limited response to the therapy, an unusual pattern of mutations was found: the insertion of 2 amino acids between residues 69 and 70, a region critical for resistance to nucleoside analogs. The higher efficacy of the combination of HU and ddI compared with that of ddI monotherapy cannot be attributed to a delayed or decreased onset of resistance to ddI. PMID- 9333148 TI - Long-term evaluation of cellular immunity during antiretroviral therapy and immunization with human immunodeficiency virus type 1 (HIV-1) Env glycoprotein in HIV-1-infected persons. AB - Cellular immune responses to human immunodeficiency virus type 1 (HIV-1) antigens, microbial recall antigens, and CD3 monoclonal antibody were studied in HIV-1-infected asymptomatic patients in a phase II, double-blind trial of immunization with recombinant HIV-1 gp160 in or not in association with zidovudine. A vigorous and persistent lymphoproliferative response (LPR) to HIV-1 Env antigens was observed in vaccinated patients. Neither Env-specific lymphocyte cytotoxicity nor LPR to recall antigens was significantly influenced by gp160 administration. The induction of LPRs to HIV-1 envelope proteins did not show positive effects on the course of HIV-1 infection. Patients treated with zidovudine alone or in combination with the immunogen showed improvement of T lymphocyte responses and transient reduction of viremia. These results suggest that antiretroviral therapy is more beneficial than immunization with gp160 and should always be considered in association with future vaccination and immunotherapeutic interventions in HIV-1-infected subjects. PMID- 9333149 TI - Effects of intravenous immunoglobulin in vivo on abnormally increased tumor necrosis factor-alpha activity in human immunodeficiency virus type 1 infection. AB - The effect of a single bolus injection (0.4 g/kg) of intravenous immunoglobulin (IVIG) on the tumor necrosis factor (TNF) system in human immunodeficiency virus type 1 (HIV-1)-infected patients was investigated. At 140 h after infusion, there was a significant decrease in levels of TNF-alpha and a significant increase in levels of soluble TNF receptors (sTNFR) in both plasma and lipopolysaccharide stimulated peripheral blood mononuclear cells (PBMC). A rapid (within 1 h) decline in expression of membrane-bound TNF-alpha and p55-TNFR on PBMC persisted throughout the study. In contrast, there was an increased expression of membrane bound p75-TNFR after 140 h. IVIG administration also resulted in significantly increased numbers of circulating CD4 lymphocytes, correlated with down-regulation of TNF-alpha activity in PBMC supernatants. Thus, down-regulation of the abnormally increased TNF-alpha activity may be achieved by IVIG administration. Studies evaluating the possible therapeutic role of long-term TNF-alpha suppression by IVIG may be warranted in HIV-1-infected patients. PMID- 9333150 TI - Neutralizing antibody responses to human immunodeficiency virus type 1 in primary infection and long-term-nonprogressive infection. AB - The role of neutralizing antibodies in human immunodeficiency virus type 1 (HIV 1) infection is poorly understood and was assessed by evaluating responses at different stages of infection. Undiluted sera from long-term nonprogressors (LTNP) had broad neutralizing antibodies against heterologous primary isolates and were more likely to neutralize the contemporaneous autologous isolate than were sera from short-term nonprogressors and progressors. In primary infection, envelope-specific IgG was detected before the initial decline in plasma viremia, but neutralizing antibodies developed more slowly. Here, neutralizing antibodies against strains SF-2 and MN were sometimes the first to be detected, but titers were low for at least 17 weeks from onset of symptoms. Neutralizing antibodies against the early autologous isolate were detected for 4 patients by 5-40 weeks but were undetectable in 2 additional patients for 27-45 weeks. The results indicate that neutralizing antibody responses are slow to develop during primary infection and are uniquely broad in LTNP. PMID- 9333151 TI - In vitro characterization of adult primary human immunodeficiency virus type 1: demonstration of distinctive single-cell killing phenotypes in spite of similar levels of viral replication. AB - Two primary human immunodeficiency virus (HIV)-1 biologic clones have been studied extensively in a system using CD4 T cell-enriched peripheral blood lymphocytes and anti-CD4 antibody to measure viral replication kinetics and single-cell cytopathicity. Biologic clones from a person with AIDS replicated to high levels and were cytopathic in the absence of syncytium formation. Unexpectedly, biologic clones from an adult long-term nonprogressor were noncytopathic in spite of similar levels of viral replication. A correlation has recently been demonstrated between reduced mitochondrial viability and cell death in HIV-1-infected cultures. Peripheral blood-derived CD4 T cells infected with the cytopathic clone showed a progressive reduction in mitochondrial viability, while those infected with the noncytopathic clone demonstrated functionally viable mitochondria. These studies demonstrate that primary HIV-1-induced cytopathicity is separable from syncytium formation and replication rate. PMID- 9333152 TI - Factors associated with increased levels of human immunodeficiency virus type 1 DNA in semen. AB - Human immunodeficiency virus type 1 (HIV-1)-infected cells have been isolated from semen and may be a major source of transmissible virus. Quantitative polymerase chain reaction (PCR) assay was used to determine HIV proviral DNA load in cellular fractions of semen from 74 antiviral therapy-naive HIV-1-seropositive men and 53 paired blood samples. HIV-1 DNA was detected in 65% of semen (range: <10-5000 copies/mL) and 100% of blood samples (range: 20-2500 copies/mL). HIV-1 DNA copy numbers in semen correlated significantly with those in blood, but for most cases, the concentration of blood HIV-1 DNA was higher (mean blood-to-semen ratio = 2.9). Factors associated with elevated HIV-1 provirus levels in semen included reduced peripheral CD4 cell count and asymptomatic genital tract inflammation (>10(6) white blood cells/mL of semen). These data provide evidence that genital tract inflammation and reduced peripheral CD4 cell count may be associated with enhanced sexual transmission of HIV-1 because of increased numbers of HIV-1-infected cells in semen. PMID- 9333153 TI - Superinfection with human immunodeficiency virus type 2 can reactivate virus production in baboons but is contained by a CD8 T cell antiviral response. AB - An animal model was used to assess whether resistance to superinfection by human immunodeficiency virus (HIV) can exist in vivo. Asymptomatic baboons (Papio cynocephalus), previously infected with HIV-2, were first challenged with homologous virus (HIV-2UC2 or HIV-2UC14) and later with heterologous virus (HIV 2UC12). After both virus inoculations, either resistance to viral infection or a transient viremia was observed. The original virus was recovered in 3 baboons, suggesting that reactivation of a latent infection occurred on heterologous challenge and that HIV-2 superinfection is blocked by processes established during prior infection. Antibody titers measured by ELISA and virus neutralization remained at low levels. However, suppression of HIV-1 replication was observed with CD8 T cells and filtered cell culture supernatants. The soluble factor involved was not a beta-chemokine. This resistance to HIV superinfection appears to be mediated at least in part by CD8 T cells that suppress virus production. PMID- 9333154 TI - T lymphocytes and macrophages, but not motile spermatozoa, are a significant source of human immunodeficiency virus in semen. AB - The cellular fraction of semen contains spermatozoa, immature germ cells, leukocytes, and epithelial cells. Recent evidence implicates seminal cells as a major source of sexually transmitted human immunodeficiency virus (HIV) in semen, but the identity and infectious potential of infected cells remains poorly understood. HIV provirus was found in 75% of viable semen cell samples by polymerase chain reaction and in 88% of paired blood cell samples from HIV seropositive men. When semen cell subpopulations were isolated by an immunomagnetic bead technique, T cells were found to be most commonly HIV infected (75% of samples), followed by macrophages (38% of samples). Viral DNA was never detected in motile spermatozoa or immature germ cell populations. Semen leukocytes proliferated in response to mitogenic and antigenic challenge and produced p24 following stimulation with irradiated allogeneic cells. These data provide evidence that both T cells and macrophages, but not germ cells, are cellular vectors of HIV transmission in semen. PMID- 9333155 TI - Clinical cryptosporidiosis and human immunodeficiency virus (HIV)-induced immunosuppression: findings from a longitudinal study of HIV-positive and HIV negative former injection drug users. AB - The natural history of cryptosporidiosis was investigated during a waterborne outbreak among 1731 members of a drug rehabilitation community in Italy; 19.6% of the members were positive for human immunodeficiency virus (HIV). Demographic and clinical information and pre-outbreak serum samples were available. Clinical data were analyzed, stratifying the study population by HIV serostatus and CD4 cell count. The attack rate of clinical cryptosporidiosis was 13.6% among HIV-negative individuals and 30.7% among HIV-positive individuals, although in the latter, it varied according to CD4 cell count. Clinical symptoms and their duration were also related to CD4 cell count. Chronic symptoms were observed in only 16 individuals (15.4%), who all had <150 CD4 cells at the onset of the illness. Among a systematic sample of 198 individuals, 14.1% already had anti Cryptosporidium antibodies before the outbreak, and 51.2% developed specific antibodies during the outbreak. The development and clinical manifestations of cryptosporidiosis were strongly influenced by the level of HIV-induced immunosuppression. PMID- 9333156 TI - Genetic similarity among Mycobacterium avium isolates from blood, stool, and sputum of persons with AIDS. AB - Large-restriction-fragment pattern comparison of Mycobacterium avium from 85 blood, stool, and respiratory specimens from 25 human immunodeficiency virus infected San Francisco patients revealed 4 strains that infected multiple people (3 groups of 2 patients and 1 group of 3 patients). Most patients harbored a single M. avium strain, but 2 strains were recovered from 8 patients. The significance of recovering 2 strains is not clear, since the second strain was seldom recovered more than once. The strain recovered from blood was recovered from stool of 4 patients and respiratory secretions of 6 patients >4 weeks before detection of bacteremia, indicating that the intestinal and respiratory tracts are entry portals from which M. avium can disseminate. M. avium from 21 cities outside of California served as controls. Thus, a single M. avium strain can cause disseminated infection in multiple patients. This may represent infection from a common environmental source or person-to-person spread. PMID- 9333157 TI - Interaction of von Willebrand factor with Staphylococcus aureus. AB - Intravascular infection due to Staphylococcus aureus requires colonization of subendothelium in the presence of shear forces. von Willebrand factor (VWF) is a large multimeric glycoprotein playing a key role in platelet adhesion to subendothelium. To determine whether VWF may also play a role in adhesion of S. aureus to endovascular sites, binding of VWF to S. aureus and adhesion of S. aureus to VWF-adsorbed substrates was examined. Binding isotherms revealed a dose dependent reaction of purified VWF with S. aureus Cowan 1 as well as VWF binding to other S. aureus strains. On solid phase, VWF showed saturable adsorption kinetics to polymethylmethacrylate and promoted S. aureus adhesion up to 67-fold in a trypsin-sensitive reaction. Similar adhesion promotion was observed when recombinant VWF was used. These results show that VWF interacts with S. aureus in suspension and promotes S. aureus adhesion to surfaces, suggesting a role of VWF in the pathogenesis of intravascular S. aureus infections. PMID- 9333158 TI - Invasive group A streptococcal infections in North Carolina: epidemiology, clinical features, and genetic and serotype analysis of causative organisms. AB - During 1994 and 1995, an increase in the number and severity of group A streptococcal (GAS) infections was noted in North Carolina. Ninety-six patients had GAS recovered from blood and other sterile body fluids, abscesses, and soft tissue. The overall case fatality rate was 11% but was much higher in patients with toxic shock syndrome (55%) and necrotizing fasciitis (58%). Recent invasive GAS isolates were compared with pre-1994 invasive isolates and temporally related pharyngeal isolates by M protein serotyping, pulsed field gel electrophoresis (PFGE), and polymerase chain reaction amplification of the streptococcal pyrogenic exotoxin A gene. Serotypes M1 and M3 accounted for 50% of recent invasive isolates (1994-1995) and 58% of pharyngeal isolates (1994). The latter isolates demonstrated PFGE patterns that were identical to invasive M1 and M3 strains, suggesting that pharyngeal infections may have served as a reservoir for virulent GAS clones. PMID- 9333159 TI - A global gene pool for high-level cephalosporin resistance in commensal Streptococcus species and Streptococcus pneumoniae. AB - Highly penicillin- and cephalosporin-resistant Streptococcus mitis and Streptococcus oralis were isolated in Spain, Hungary, and Berlin. With chromosomal DNA of these strains, resistant transformants of Streptococcus pneumoniae were obtained that expressed low-affinity variants of penicillin binding proteins (PBPs) 2x, 1a, 2a, and 2b in different combinations, depending on the selective conditions. The transformants had cefotaxime MICs of up to 6 microg/mL, and those with a low-affinity PBP 2b were highly deficient in penicillin-induced lysis. Sequence analysis of the pbp2x genes confirmed the presence of a global gene pool of penicillin resistance determinants shared by commensal and pathogenic streptococci. PMID- 9333160 TI - Utility of a polymerase chain reaction diagnostic system in a study of the epidemiology of shigellosis among dysentery patients, family contacts, and well controls living in a shigellosis-endemic area. AB - Polymerase chain reaction (PCR) diagnostic methods have rarely been used in epidemiologic studies of Shigella and enteroinvasive Escherichia coli (EIEC) infections. In this study, amplification of the invasion plasmid antigen H (ipaH) gene by PCR and standard culture methods was used to identify Shigella species or EIEC among 154 patients with dysentery, 154 age-matched controls, and family contacts in Thailand. The ipaH PCR system increased the detection of Shigella species and EIEC from 58% to 79% among patients with dysentery and from 6% to 22% among 527 family contacts; 75% of infections in family members were asymptomatic. Detection of the ipaH gene was statistically associated with dysentery. Household contacts of patients with shigellosis diagnosed only by PCR had significantly higher rates of shigellosis than household contacts of patients who did not have Shigella or EIEC infections. Detection of the ipaH gene by PCR is far more sensitive than detection by standard culture and is highly correlated with evidence of Shigella transmission among family contacts. PMID- 9333161 TI - Antibody-mediated depletion of tumor necrosis factor-alpha impairs pulmonary host defenses to Legionella pneumophila. AB - Tumor necrosis factor-alpha (TNF-alpha) has been shown to stimulate the resistance of alveolar macrophages and neutrophils to Legionella pneumophila in vitro. To determine whether endogenous TNF-alpha is necessary for host defense against legionellosis in vivo, anti-TNF-alpha IgG or control IgG was administered to rats exposed to aerosolized L. pneumophila. Treatment with anti-TNF-alpha neutralized >90% of the intrapulmonary TNF-alpha response to infection, resulting in persistent pneumonitis and failure to clear L. pneumophila from the lungs. Depletion of TNF-alpha limited the recruitment of mononuclear cells to the lungs and resulted in a progressive increase in the proportion of alveolar macrophages that were infected; neutrophil recruitment and phagocytosis were not impaired. Both systemic and intrapulmonary IFN-gamma levels were significantly higher in rats depleted of TNF-alpha. These observations indicate that TNF-alpha is required for the prompt resolution of pneumonic legionellosis and point to a direct role for TNF-alpha in the activation of phagocytes. PMID- 9333163 TI - DNA vaccination with the major outer-membrane protein gene induces acquired immunity to Chlamydia trachomatis (mouse pneumonitis) infection. AB - The efficacy of DNA vaccination for prevention of Chlamydia trachomatis infection was studied using the murine model of pneumonia induced by the mouse pneumonitis (MoPn) isolate of C. trachomatis. Intramuscular DNA immunization with two chlamydial genes, one that encodes the major outer-membrane protein (MOMP) and one that encodes a cytoplasmic enzyme (cytosine triphosphate [CTP] synthetase) were tested. The MOMP DNA vaccine but not the CTP synthetase DNA vaccine generated significant delayed-type hypersensitivity and serum antibodies to MoPn elementary bodies and reduced the peak growth of MoPn by >100-fold following lung challenge infection. MOMP DNA immunization suggests a new approach to vaccine development for prevention of human chlamydial infection. PMID- 9333162 TI - Antigenic diversity of granulocytic Ehrlichia isolates from humans in Wisconsin and New York and a horse in California. AB - The agent of human granulocytic ehrlichiosis (HGE), Ehrlichia phagocytophila, and Ehrlichia equi are very similar. HGE is of variable severity. Genetic and antigenic differences among 3 human isolates (Webster, Spooner, and NY-8) and 1 horse isolate (MRK) were evaluated. The 16S rRNA gene sequences were identical in all human isolates. By use of 5 homologous antisera from these 3 humans and 1 horse and an additional 5 antisera in heterologous reactions, the immunodominant antigens of each isolate were noted to differ in molecular size: 43 kDa in the Webster (Wisconsin) isolate, 46 kDa in the Spooner (Wisconsin) isolate, 42 and 45 kDa in the NY-8 (New York State) isolate, and a 42 kDa doublet in the E. equi MRK isolate from California. Two sera from a Wisconsin patient reacted weakly or not at all with the NY-8 isolate. Antigenic structural diversity exists among otherwise indistinguishable granulocytic ehrlichial isolates. PMID- 9333164 TI - Monoclonal antibody-mediated, complement-independent binding of human tumor necrosis factor-alpha to primate erythrocytes via complement receptor 1. AB - Monoclonal antibody (MAb)-based heteropolymers (HP) were used to simulate immune adherence. The HP is constructed by cross-linking MAbs that recognize complement receptor 1 (CR1) and tumor necrosis factor-alpha (TNF-alpha). 125I-labeled TNF alpha was cocultured with either sheep, monkey, or human erythrocytes in the presence or absence of HP. Human erythrocytes demonstrated 63% +/- 0 (mean +/- SD) binding of 125I-labeled TNF-alpha, while binding of 125I-labeled TNF-alpha in the absence of HP was 4% +/- 1% (P < .001). Monkey erythrocytes showed similar results, while sheep erythrocytes (which lack CR1) demonstrated low binding. The effect of HP binding on biologic activity of TNF-alpha was examined in an assay of stimulated human neutrophils. The HP completely inhibited the ability of TNF alpha to prime neutrophils, occurring regardless of the presence or absence of erythrocytes but solely dependent on the addition of HP. Thus, the HP facilitated specific, saturable, and significant binding of 125I-labeled TNF-alpha to primate erythrocytes in vitro. PMID- 9333165 TI - Seroepidemiology of Trypanosoma cruzi, etiologic agent of Chagas' disease, in US blood donors. AB - A comprehensive seroepidemiologic study was conducted in two Red Cross regions (Los Angeles and Miami) to determine the prevalence of Trypanosoma cruzi antibodies in at-risk blood donors, to identify additional risk factors, and to assess the likelihood of transmitting T. cruzi by transfusion. At-risk and control donors were stratified by a broad risk question, tested for T. cruzi antibodies, and if confirmed as seropositive, enrolled in case-control and lookback investigations. A total of 299,398 donors were queried; 23,978 at-risk and 25,587 control donations were tested, and T. cruzi antibodies were confirmed in 34 donors (33 and 1, respectively). Seropositive donors shared one risk factor; birth/extensive time in a T. cruzi-endemic area. Lookback studies identified 11 recipients, all negative for T. cruzi antibodies. Screening strategies that use a question are unlikely to identify all seropositive donors. The lack of definitive data on the risk of transmission by transfusion indicates additional studies of donors and recipients are needed. PMID- 9333166 TI - Perspective: aging and infectious diseases: past, present, and future. AB - As we enter into the 21st century, infectious disease specialists will be managing a greater number and proportion of patients with infections who are > or = 65 years old. Much has been learned about aging, host resistance, and infections over the past 15 years. However, if we are to meet the challenge of the complex issues of geriatric infectious diseases, infectious disease clinicians, teachers, and researchers must assume a more proactive role in clinical care, training, education, and research on problems and issues confronting the aging population. PMID- 9333167 TI - International Colloquium on Ebola Virus Research: summary report. PMID- 9333168 TI - Response to hepatitis A vaccination in human immunodeficiency virus-infected and uninfected homosexual men. AB - The influence of human immunodeficiency virus (HIV) infection and vaccination schedule on the immunogenicity of a hepatitis A vaccine was examined. Ninety HIV infected homosexual men received two vaccinations with hepatitis A vaccine (each 2 mL of 720 ELISA units/mL) either 1 or 6 months apart; 44 HIV-uninfected men received vaccine at study entry and at 6 months. Anti-hepatitis A virus (HAV) titer after vaccination was measured in 83 HIV-positive and 39 HIV-negative men. Seroconversion (anti-HAV antibody > or = 20 IU/L) after two vaccinations occurred more frequently in HIV-negative men (100% vs. 88.2%; P = .03). Anti-HAV titer after two vaccinations was also significantly greater in HIV-negative men (1086 vs. 101 IU/L; P = .0001). HIV-positive men who responded to vaccination had significantly more CD4 lymphocytes (mean, 540/microL) at baseline than those who did not (280/microL; P = .033). Vaccine schedule did not affect response. Vaccination of susceptible patients against HAV should be recommended early in HIV infection using the shorter course to encourage compliance. PMID- 9333169 TI - Dominant cytotoxic T lymphocyte response to the immediate-early trans-activator protein, BZLF1, in persistent type A or B Epstein-Barr virus infection. AB - Five healthy human leukocyte antigen-B8 (HLA-B8)-positive virus carriers were studied to investigate the CD8+ cytotoxic T lymphocyte (CTL) response to an HLA B8-restricted peptide, RAKFKQLLQ, located in the Epstein-Barr virus (EBV) immediate-early trans-activator protein, BZLF1. Of the 5 virus carriers, 4 were infected with type A and 1 with type B EBV. Using limiting-dilution analysis of peripheral blood mononuclear cells, a high RAKFKQLLQ-specific CTL precursor frequency was demonstrated after specific peptide or autologous lymphoblastoid cell line stimulation in both type A and type B EBV carriers. The RAKFKQLLQ specific CTL precursor frequencies in all 5 persons were at least as dominant as those observed with two other EBV-associated, HLA-B8-restricted latent epitopes, FLRGRAYGL and QAKWRLQTL. These findings show that healthy virus carriers maintain a high frequency of BZLF1-specific memory T cells, potentially to control virus spread from lytically infected cells. PMID- 9333170 TI - Transmission of vaccine strain varicella-zoster virus from a healthy adult with vaccine-associated rash to susceptible household contacts. AB - Twelve days after receiving an investigational Oka strain live attenuated varicella vaccine, a 38-year-old healthy white woman developed a rash consisting of 30 scattered lesions. Sixteen days later, her 2 children also developed rash. Swabs obtained from the skin lesions of the vaccinee and her children demonstrated the presence of varicella-zoster virus (VZV) DNA by a polymerase chain reaction (PCR) assay. Restriction endonuclease polymorphisms present in wild and vaccine type VZV were examined, and the amplified VZV DNA was determined to be vaccine type. This case documents transmission of varicella vaccine type virus from a healthy vaccinee to susceptible household contacts. Since vaccine associated rashes are uncommon and mild, it is likely that transmission of vaccine virus will also be uncommon. With widespread immunization beginning in the United States, ongoing studies will define the frequency of this transmission. PMID- 9333171 TI - Papillomavirus is resistant to desiccation. AB - There is strong epidemiologic evidence for sexual transmission of high-risk genital human papillomavirus (HPV) types. However, it is unclear if infection may also be transmitted indirectly via fomites. To assess this possibility, the in vitro infectivity after desiccation was compared for pseudotype HPV-16 virions, a model for high-risk type genital HPV, and bovine papillomavirus type 1 (BPV-1), a papillomavirus known to be transmitted via fomites. The 2 viruses had similar resistance to desiccation in cell extracts, retaining approximately 100%, 50%, and 30% of infectivity when dehydrated for 1, 3, and 7 days, respectively, at room temperature. Pseudotype HPV-16 and BPV in cell extracts were completely inactivated by autoclave treatment and susceptible to 70% ethanol but were resistant to EDTA or incubation at 56 degrees C for 1 h. The data suggest that further study of nonsexual spread of high-risk genital HPV via fomites is warranted. PMID- 9333172 TI - Polymerase chain reaction detection and clinical significance of varicella-zoster virus in cerebrospinal fluid from human immunodeficiency virus-infected patients. AB - Varicella-zoster virus (VZV) causes ocular and other central nervous system (CNS) disease in human immunodeficiency virus (HIV)-infected persons. To study the prevalence of CNS disease due to VZV, cerebrospinal fluid (CSF) specimens from 84 consecutive HIV-infected patients with new neurologic symptoms were tested for VZV DNA by a polymerase chain reaction (PCR) assay. Six patients were PCR positive for VZV in CSF; 3 additional patients were subsequently identified who were not part of the serial population sample. Among these 9 patients, all had clinical presentations consistent with ocular and other CNS disease due to VZV; 4 were without zoster on presentation. Sustained improvement in association with antiviral therapy was observed in 3. Therefore, VZV DNA was detected in the CSF of 7% of HIV-infected patients presenting with neurologic symptoms; the diagnosis of VZV-related CNS disease was facilitated by this assay; improvement in association with antiviral therapy was observed in some patients. PMID- 9333173 TI - Delayed-type hypersensitivity skin testing using third variable loop peptides identifies T lymphocyte epitopes in human immunodeficiency virus-infected persons. AB - Cellular immune responses to human immunodeficiency virus type 1 (HIV-1) infection, particularly in vivo responses, have been difficult to study in large patient cohorts because of technical impediments. By use of small peptide fragments of the HIV-1 gp120 third variable loop, the CD4 T lymphocyte epitopes of 2 HIV-infected persons were mapped using a cutaneous delayed-type hypersensitivity (DTH) assay. The in vivo DTH responses correlated with epitopes previously identified in vitro using CD4 T lymphocyte lines. The ability to determine CD4 T lymphocyte epitopes in large cohorts of patients using this simple in vivo technique would provide important diagnostic and prognostic data regarding effective immunoregulation of HIV-1. This technique should have broad applicability in HIV vaccine development and in the investigation of other immune mediated human diseases. PMID- 9333174 TI - Is crusted (Norwegian) scabies a marker of adult T cell leukemia/lymphoma in human T lymphotropic virus type I-seropositive patients? AB - Human T cell lymphotropic virus type I (HTLV-I)-induced immunosuppression has been suggested to explain the occurrence of crusted scabies in HTLV-I-infected patients. HTLV-I is the etiologic agent of adult T cell leukemia/lymphoma (ATL). Crusted scabies diagnosed in 6 HTLV-I-seropositive patients was studied to look for an association with ATL. Four of the 6 either had concomitant ATL when crusted scabies was diagnosed or developed ATL a few months later. These findings suggest that the occurrence of crusted scabies in patients seropositive for HTLV I could represent a sign of marked immunosuppression related to ATL. PMID- 9333175 TI - CCR5 genotypes in sexually active couples discordant for human immunodeficiency virus type 1 infection status. AB - Persons who are homozygous for the delta32 polymorphism of the CCR5 chemokine receptor gene are highly protected against human immunodeficiency virus type 1 (HIV-1) infection. Previous studies described 54 HIV-1-discordant couples in whom no virus transmission occurred despite extensive sexual contact. The possible role of the delta32 polymorphism in the lack of HIV-1 transmission between these partners was studied. No participants were homozygous for the delta32 allele, but the proportion that was heterozygous was higher among HIV-1-seronegative than HIV 1-seropositive partners (28% vs. 11%, P = .05). This association was seen in heterosexual couples (P = .03) but not in homosexual couples (P = .74). Among white persons, who are most likely to carry the delta32 allele, 38.9% of HIV-1 uninfected and 5.6% of HIV-1-infected heterosexual partners were heterozygous (P = .04). These data are consistent with a possible association between the heterozygous delta32 genotype in heterosexual sex partners and partial protection against HIV-1 infection, and they emphasize the importance of analyzing different risk groups in studies of host factors that influence infection. PMID- 9333176 TI - Expression of collagen-binding protein and types 5 and 8 capsular polysaccharide in clinical isolates of Staphylococcus aureus. AB - In vitro collagen binding of 216 Staphylococcus aureus isolates from patients with various diagnoses was studied. Polymerase chain reaction was used to examine these isolates regarding the existence of the corresponding cna gene. Distribution of capsular polysaccharide (CP) types was examined. Fifty-six (57%) of 99 S. aureus isolates from patients with endocarditis or bacteremic bone or joint infection were cna-positive compared with 65 (56%) of 117 isolates from bacteremic patients without signs of bone or joint infection (P = .99). There was a good correlation between in vitro collagen binding and presence of the cna gene. These data suggest that collagen binding is not a prerequisite for the development of endocarditis, osteomyelitis, or septic arthritis. There was no significant difference in the distribution of CP types among various patient groups, although there was a strong association between CP type 8 and the existence of the cna gene. PMID- 9333177 TI - Persistence of antibodies to pneumococcal capsular polysaccharide vaccine in the elderly. AB - Persistence of antibodies to 23-valent pneumococcal vaccine was assessed among 62 subjects aged 65-88 years. IgG antibodies were measured by standardized EIA to serotypes 4, 6B, 9V, 14, 19F, and 23F before and 1 month, 1 year, and 3 years after vaccination. After satisfactory antibody responses (fold increases from 2.6 to 5.3), 3-year geometric mean concentrations (GMCs) had waned to close (for types 4, 9V, and 23F) or similar (for types 6B and 19F) to their prevaccination values. Type 14 was exceptional: 1-month GMC was 7.7-fold and 3-year GMC was 3.0 fold in comparison to the prevaccination GMC. Antibody concentrations decreased at an equal rate irrespective of serotype and age or sex of the vaccinee. The major factor predicting the persistence of antibodies above the prevaccination level was the magnitude of the original antibody response. Present results suggest that pneumococcal revaccination of the elderly may be needed as early as 3-4 years after the initial vaccination. PMID- 9333178 TI - Restriction fragment length polymorphism analysis and random amplified polymorphic DNA analysis of Campylobacter jejuni strains isolated from patients with Guillain-Barre syndrome. AB - Campylobacter jejuni serotype O19 strains associated with the Guillain-Barre syndrome (GBS) and other strains were examined by restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction products of the flaA genes and by random amplified polymorphic DNA (RAPD) analysis. RFLP analysis showed that regardless of LIO serotype, geographic origins, or association with GBS, the O19 isolates shared an identical digestion pattern by each of four restriction endonucleases, DdeI, MboI, MseI, and AluI. In contrast, among C. jejuni O1 or O2 strains, RFLP patterns were different even among strains of the same LIO serotype. The results of the RAPD analysis were consistent with the flaA RFLP data. These data indicate that all of the O19 strains that were tested were closely related to one another whether they were or were not associated with GBS. PMID- 9333179 TI - In vitro efficacy of antimicrobial-coated bladder catheters in inhibiting bacterial migration along catheter surface. AB - Most cases of catheter-related urinary tract infection are probably caused by organisms that migrate from the urethral meatus-catheter interface along the external surface of the catheter into the bladder. To examine the ability of bladder catheters coated with minocycline and rifampin to inhibit bacterial migration along the external surface of the catheter, a novel in vitro bladder model was used. Compared with uncoated catheters, antimicrobial-coated bladder catheters significantly impeded the migration of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterococcus faecalis, and Candida albicans (bacteriuria developed at a mean of 2-5 days vs. 9-34 days, respectively, after bacterial contamination of the catheter). Although production of zones of inhibition by coated catheters may provide some protection against infection, there was no correlation between the size of zones of inhibition and level of efficacy in inhibiting bacterial migration in vitro. Examination of the clinical efficacy of these antimicrobial-coated bladder catheters is prudent. PMID- 9333180 TI - Treatment of African children with uncomplicated falciparum malaria with a new antimalarial drug, CGP 56697. AB - New antimalarial drugs are urgently needed. The use of short courses of the new antimalarial drug artemether as monotherapy has been limited by secondary malaria episodes following parasite clearance. Therefore, a new antimalarial drug, CGP 56697, has been developed, which combines artemether with a longer-acting antimalarial agent, benflumetol. A safety trial was undertaken in 60 Gambian children 1-6 years old with uncomplicated Plasmodium falciparum malaria. All children treated with CGP 56697 cleared their parasites 72 h after the start of treatment. No neurologic, cardiac, or other adverse reactions were observed. Second episodes of falciparum malaria were recorded in 16 (27%) of the children. Second infections were more frequent during the rainy season than during the dry season. Molecular epidemiologic studies suggested that 12 of the 14 second episodes of malaria in children treated with CGP 56697 were due to new infections. CGP 56697 proved to be a safe and effective antimalarial drug in African children. PMID- 9333181 TI - Response to interferon-gamma plus pentavalent antimony in Indian visceral leishmaniasis. AB - One hundred fifty-six previously untreated Indian patients with visceral leishmaniasis were treated with pentavalent antimony (Sb) alone for 30 days (group A), Sb plus interferon-gamma (IFN-gamma) for 30 days (group B), or Sb plus IFN-gamma for 15 days (group C). The purpose was to show that IFN-gamma would increase the response to 30 days of Sb treatment and that short-course (15 days) combination therapy was as effective as 30 days of Sb alone. Six months after treatment, 36% of group A, 49% of group B, and 42% of group C patients were designated as definitively cured. The success rates for long-term responses to Sb alone (36%) and Sb plus IFN-gamma (49%) were unexpectedly low, and responses in groups A, B, and C were not significantly different. These results suggest that the beneficial effects of adjunctive IFN-gamma in visceral leishmaniasis may be limited in regions where this disseminated intracellular infection shows high level resistance to Sb. PMID- 9333182 TI - Cryptosporidiosis in adults in Lusaka, Zambia, and its relationship to oocyst contamination of drinking water. AB - In Lusaka, where human immunodeficiency virus seroprevalence in young adults is approximately 25%, four townships were studied to establish the prevalence of persistent diarrhea in adults and the etiologic importance of cryptosporidiosis in adults with persistent diarrhea. Cryptosporidium parvum oocyst contamination of urban water supplies was measured and the results used to categorize these populations into high or low exposure. In total, 506 adults were reported as having had diarrhea in the 2 weeks prior to the survey; 101 of these episodes were persistent. Adults with persistent diarrhea in the high-exposure areas were more likely to have cryptosporidiosis (odds ratio, 5.14; 95% confidence interval, 1.57-17.2; risk ratio, 1.83; 95% confidence interval, 1.04-3.21; P = .003) although overall prevalence of persistent diarrhea was not greater in these areas. This association was not confounded by animal exposure, travel, or boiling water. Within these urban populations, water contamination with C. parvum was a major influence on the prevalence of infection. PMID- 9333183 TI - Polymerase chain reaction evaluation for Mycoplasma pneumoniae. PMID- 9333184 TI - Lack of clinical significance for the common arginine-to-leucine substitution at codon 463 of the katG gene in isoniazid-resistant Mycobacterium tuberculosis in Singapore. PMID- 9333185 TI - The effects of photoperiod and food intake on reproductive development in male deer mice (Peromyscus maniculatus). AB - Seasonal breeding is a tactic that has evolved in rodents that limits reproduction to specific times of the year to increase reproductive success. In order to time breeding accurately, many animals respond to changes in daily photoperiod. Short day lengths inhibit breeding in many nontropical rodent species. Restricted food availability can also inhibit reproductive function among some individuals in these so-called "photoperiodic" populations. Rodents born at the end of the breeding season typically delay sexual maturation until the following spring. Prepubertal rodents exposed to day lengths that are < 12 h light/day will not undergo puberty for 4-7 months in the laboratory. Food restriction can also affect the timing of puberty onset. Reproductive function of food-restricted juvenile mice may remain inhibited until food availability improves. Alternatively, reproductive function of food-restricted juvenile mice might eventually develop despite restricted food intake. This study examined the effects of chronic food restriction and photoperiod on reproductive development in male deer mice (Peromyscus maniculatus bairdii). Short-day mice fed ad lib delayed gonadal development for 5-7 months, but eventually achieved reproductive maturity. The reproductive function of short-day mice fed ad lib was indistinguishable from long-day control animals when assessed at week 32. Long day food-restricted mice exhibited an intermediate level of gonadal development and function. Short-day food-restricted deer mice also inhibited reproductive growth, but failed to demonstrate reproductive maturity by week 32 of the study. Taken together, these results suggest that retardation of reproductive development by food restriction is only superficially similar to the delay in reproductive maturation imposed by short day exposure. It does not appear that male deer mice escape from the inhibitory effects of food restriction to attain sexual development. PMID- 9333186 TI - Gait analysis in a rat model of osteoarthrosis. AB - Gait analysis has been undertaken in a rat model of osteoarthrosis, induced by intra-articular injection of sodium iodoacetate into the left knee. Two weeks after injection, no disturbances were recorded to the velocity of locomotion, the stride length nor the stride, stance, or swing times. However, clear and consistent reductions in the peak vertical load bearing (Pz) by the affected limb were observed of 22-29% relative to the other limbs, with the right forelimb taking the major share of extra load. This redistribution fitted well with the gait pattern of the rat, allowing Pz redistribution with minimum gait disturbance, and was still present 6 weeks later. These results are discussed in the context of the possible load sensitivity of the damage process to the gait pattern of the rat. PMID- 9333187 TI - Altered NMDA sensitivity and learning following chronic developmental NMDA antagonism. AB - We have previously shown that chronic developmental administration of N-methyl-D aspartate (NMDA) antagonists reduces synaptic development; however, on withdrawal from NMDA antagonism, there is a rebound period during which synaptogenesis exceeds control levels. The current research was undertaken to explore this period of withdrawal, using the noncompetitive antagonist phencyclidine (PCP), examining 2 behavioral measures in which the NMDA receptor is implicated: 1. NMDA induced seizures, and 2. learning and memory in the Morris water maze. Using a protocol identical to that previously used to examine synaptic development, male Long-Evans rats were given 1 daily SC injection of either 10 mg/kg PCP or its physiological saline vehicle for a period of 15 days, beginning on postnatal Day 5 (P5) and ending on P20. Animals were then assessed for either sensitivity to NMDA-induced seizures on P21, P26, P36, or P56, or they were assessed for their acquisition performance and initial heading in the Morris water maze on P23, P26, P30, P38, and P75. Chronic treatment with PCP resulted in greater behavioral ratings of seizure activity after NMDA administration, observed 1 (P21), 5 (P26), and 15 (P36) days after the last injection of PCP, indicating increased sensitivity of the NMDA receptor/channel complex during this period after withdrawal from developmental NMDA antagonism. PCP-treated animals also required significantly more trials to reach criterion in the Morris water maze on P23, P26, and P30, and displayed significantly less accurate initial swim headings on all test days. The results are discussed in terms of the role of the NMDA receptor-channel complex in development and learning/memory processes. PMID- 9333188 TI - Open field activity and human interaction as a function of age and breed in dogs. AB - Open field (OF) activity was studied in kennel reared purebred beagles from two separate colonies (2-13 years in age) and pound source mixed breed dogs (9 months to 10 years in age). Dogs were observed for 10 min sessions and records were taken of: locomotion, urination, sniffing, grooming, rearing, vocalizing, jumping frequencies and inactivity (16). Since dogs are uniquely social towards people, we also measured human interaction (HI), which recorded the same behaviors as during OF when a person was present in the room. Measures of exploratory behavior decreased as a function of age in pound source dogs in the OF test, but not in beagles from either colony. No breed differences were found between the young dogs. In the HI test, age effects were found in beagles but not pound source dogs. OF activity correlated with tests of cognitive function, but differences were found between the three groups. These findings indicate that OF activity is age-sensitive in dogs, but that breed and test conditions are also essential factors. PMID- 9333189 TI - Adjustment of house sparrow circadian rhythms to a simultaneously applied light and food zeitgeber. AB - Periodic food availability has been shown to be an effective circadian zeitgeber in many vertebrates. It is still unclear, however, i) whether light-active species like most birds can synchronize with food cycles in the presence of a strong light-dark (LD) cycle and ii) whether it is common among non-mammalian vertebrates to use a separate circadian oscillator to synchronize with food cycles as most mammals do. We investigated these questions experimentally by exposing house sparrows simultaneously to two zeitgebers: light and food. The LD cycle was set at 1410 h; food was always available for 12 hour per day, but at different phases of the LD cycle. The effects of the two zeitgebers were analyzed by observing two behavioral outputs of the birds' circadian system, the rhythms of locomotion and feeding. The data revealed that light acted as the dominant zeitgeber in most conditions. Food cycles affected the phase of the behavioral rhythms of the birds only when the food was presented no later than 3 h after the onset of light. Apart from their synchronizing actions both light and food cycles also exerted direct (masking) effects on the behavioral rhythms of the birds. The results suggest that the circadian system of house sparrows can indeed adjust to two simultaneous environmental periodicities, i.e. light and food. We propose that light is the stronger zeitgeber and plays a 'permissive' role in determining the phases at which synchronization with food cycles comes into effect. We did not find evidence that the house sparrows' behavioral rhythms are controlled by a food-entrainable circadian oscillator that is distinct from the light-entrainable system as is the case in most mammals. The differences in the patterns of food synchronization and organization of the circadian system that appear to exist between different species can be interpreted in two ways: i) species of different phylogenetic origin (e.g., mammals versus birds) evolved different circadian system or ii) regardless of phylogeny, species with different ecological requirements show a specialization in their circadian organization which is adjusted to the importance of zeitgebers in nature. PMID- 9333190 TI - Manipulation of behavioral disorders in autoimmune mice via prolactin. AB - Autoimmune mice perform poorly in two-way active avoidance tasks, and the extent of this performance deficit appears to be related to the extent of autoimmunity following developmental manipulations. In the current study, the pituitary hormone prolactin, which has immune-enhancing effects, was used to manipulate this behavioral disorder in adulthood. Prolatinergic manipulation may be achieved by the use of dopaminergic drugs. In two experiments, autoimmune NZB X NZW F1 (BW) mice received either pimozide (PIM; a D2 antagonist) or bromocriptine (CB154; a dopamine agonist) in their drinking water. Control subjects received plain water. Following treatment, subjects were tested in an activity monitor, and active avoidance learning. Circulating PRL levels, as measured by RIA, were significantly increased by PIM and significantly decreased by CB154. Neither drug affected circulating levels of autoantibodies to DNA or cardiolipin, a phospholipid. In Experiment 1, in which mice were tested at 12 weeks of age, after 6 weeks of drug treatment, PIM treated animals of both sexes showed significantly more failures to escape the shock in avoidance conditioning, while CB154 did not have significant effects. In Experiment 2, in which mice were tested at 16 weeks of age, after 12 weeks of drug treatment, CB154 treated females (males were not tested) showed significantly fewer failures to escape, while PIM did not have significant effects. The effects of PRL on behavior, and its relation to immune system function, are discussed. PMID- 9333191 TI - Visuospatial discrimination deficit in rats after ibotenate lesions in anteromedial visual cortex. AB - To assess the role of the rat anteromedial extrastriate cortex (AM) in a visuospatial discrimination task, restricted bilateral ibotenic acid lesions were placed stereotaxically in this region. Gray rats with lesions in AM were trained in a task requiring them to discriminate the location of a light stimulus placed vertically at different elevations. Correct responses required pressing right or left levers to obtain rewards. In contrast to unoperated controls, lesioned rats failed in learning the visuospatial discrimination task. A correlation was found between the bilateral extent of the lesion in area AM and the behavioral deficit. Another group of lesioned rats was trained to discriminate brightness differences of the light stimulus but requiring the same egocentric right/left motor response. The performance of these rats was similar to that of controls. From these results we conclude that extrastriate area AM in the rat is necessary for visuospatial discrimination, but not for correct egocentric motor responses. The visuospatial functions of area AM in the rat are reminiscent of visuospatial functions ascribed to the parietal streams of extrastriate visual areas in the monkey. PMID- 9333192 TI - Calories affect zeitgeber properties of the feeding entrained circadian oscillator. AB - Rats with suprachiasmatic (SCN) lesions readily entrain to daily meals by increasing their activity prior to food access. Although previous experiments suggest that entrainment requires a nutritive meal, it is not yet clear what the parameters of the entraining stimulus are. The first experiment investigated the role of caloric content in resetting the feeding entrainable oscillator (FEO). Rats were entrained to 20 g chow/day until anticipatory activity was stable. The food access time was then delayed by 8 h and rats received either 0, 2, 6, or 16 g of chow for two days. Sixteen g of chow produced large delays on the next two cycles, while 0 and 2 g produced no delays. Two of 8 rats receiving 6 g showed delays, indicating that 22 kcal is near the threshold. In a second experiment, the effects of bulk were investigated. After an 8 h phase delay, rats received 0, 6, 10 or 16 g of chow mixed with cellulose for a total of 21 g for all groups. The 10 and 16 g chow groups delayed while the 0 g chow group did not. In the 6 g groups some animals phase shifted while others did not. Thus, the addition of non nutritive bulk to the phase shifted meal had little or no effect on resetting the FEO and it appears that caloric content rather than gastric distention provides an effective zeitgeber for the FEO. PMID- 9333193 TI - Effect of vomeronasal organ removal on male socio-sexual responses to female in a prosimian primate (Microcebus murinus). AB - In most mammals, olfactory cues play an important role in regulating socio-sexual behaviors, but the relative contributions of the main olfactory system and the vomeronasal system remain unclear. The lesser mouse lemur, a nocturnal prosimian, possesses well-developed chemosensory structures, including a functional vomeronasal organ (VNO). In this primitive primate, social communication and competition between males for priority access to receptive females includes reliance on urinary chemical cues. To assess the role of the VNO in mediating males' behavioural responses to females, sexually-experienced intact males (C, n = 8) or males deprived of their VNO (VNX, n = 8) were put in pairs and their socio-sexual behaviors in response to a preoestrous female were monitored. Independent of social context (with or without female), VNX males exhibited less sniffing behaviors than C males, but their marking behaviors, although reduced, were not significantly different. Removal of the VNO dramatically reduced the frequency of both sexual behaviors (anogenital investigations, mounts) and intermale aggressive behaviors. However, VNO removal did not impair successful mating and had no effect on plasma testosterone levels. Lastly, VNX males always exhibited a significantly lower general activity level than C males. The decrease in sexual behaviors and the lack of aggressive intermale competition in responses to a preoestrous female possibly proceed from functional disturbances of central nervous areas connected to the vomeronasal system rather than from a chemosensory deficit per se. PMID- 9333194 TI - Olfaction in the domestic fowl: a critical review. AB - It has been known for some time that many species of birds, including domestic fowl Gallus domesticus, have an olfactory sense. However, the functional significance of avian olfaction is less clear. We review neurobiological, embryological and behavioral evidence relevant to the question of how domestic fowl use the sense of smell. Evidence suggests a potential role for olfaction in the formation of attachments to familiar objects or environments; in the elicitation of fear responses by alarm and predator-related odors; in the control of feeding and drinking; and in avoidance of noxious substances. The fact that domestic fowl can detect and respond to a wide range of odors, in a variety of behavioral contexts, has important practical implications, especially in relation to welfare and husbandry. PMID- 9333195 TI - Nesting material as environmental enrichment has no adverse effects on behavior and physiology of laboratory mice. AB - Environmental enrichment may improve the quality of life of captive animals by altering the environment of animals so that they are able to perform more of the behavior that is within the range of the animal's species-specific repertoire. When enrichment is introduced into an animal's environment, it is important to evaluate the effect of the enrichment program and to assess whether the animal continues to use the enrichment in the long-term. Groups of mice were housed under either standard or enriched conditions for several weeks. Nesting material which was highly preferred in previous studies was used as enrichment. During the period of differential housing several behavioral parameters (behavioral tests and handling) and physiological parameters (urine and plasma corticosterone, food and water intake, body and adrenal weight) were monitored to determine the impact of environmental enrichment. Observations were made to determine whether or not the mice continued to use the enrichment. The results indicated that throughout the study all mice used the nesting material to build nests and that mice from enriched conditions weighed more than mice housed under standard conditions, although the latter consumed more food. No major differences for behavioral and physiological parameters were found between the groups of mice housed under different conditions. Therefore it is not likely that supply of nesting material will jeopardize the outcome of experiments. PMID- 9333196 TI - Temporal changes in fat pad mass and cellularity after lipectomy in Siberian hamsters. AB - Long-day-housed Siberian hamsters show increases in the mass of nonexcised white adipose tissue (WAT) 12 weeks after bilateral removal (x) of epididymal WAT (EWAT). Although EWAT shows no significant regeneration, surviving EWAT appears to increase in mass to a small degree. The purpose of the present study was to determine the time course for fat pad mass and cellularity changes after EWATx and to test whether surviving EWAT adipocytes undergo hypertrophy or hyperplasia. Male Siberian hamsters underwent EWATx or sham EWATx (SHAM). At Week 0 and at 2 week intervals up to 12 weeks postsurgery, a representative sample of animals from each group were killed. EWAT, retroperitoneal WAT (RWAT), and inguinal WAT (IWAT) were removed, weighed, and processed for cellularity measurement. IWAT and, to a nonsignificant degree, RWAT, increased in mass after EWATx due to fat cell hypertrophy. These changes began as early as Week 4 postlipectomy, but no mass or cellularity change was significant until Week 12. The surviving EWAT adipocytes of EWATx hamsters also were larger than those of SHAM hamsters and, unlike EWAT adipocyte number, increased with time within the EWATx group. SHAM hamsters showed slight increases in the mass of EWAT, but not IWAT or RWAT, due to a nonsignificant doubling of EWAT adipocyte number during the 3 months of the experiment. PMID- 9333197 TI - Vegetative background of sleep: spectral analysis of the heart rate variability. AB - Phasic events during sleep, either arousal or REM-burst related, and the associated transient cardiovascular responses have been the subject of intensive research in previous studies. However, nontransient (stationary) fluctuations in heart rate have been studied less extensively in the past. They allow a differentiation of the sympathetic and parasympathetic activation, which are related to a low-frequency (LF) and a high-frequency (HF) component of the heart rate variability (HRV) signal, respectively. The resulting LF/HF ratio is a quantitative index of the sympatho-vagal balance. Sleep polygrams from 20 healthy volunteers were recorded in a sleep laboratory. Standard vegetative tests (orthostatic and Valsalva tests) were evaluated. A segmentation procedure, performed on the HRV signal, separated 70-130 transients during the night from the records of stationary heart rate fluctuations. From these periods the sympatho-vagal balance, quantified by the LF/HF, was computed by means of spectral analysis. The more synchronized the sleep was, the more the LF/HF decreased, whereas the LF/HF was significantly increased during REM sleep, indicating a sympathetic predominance during this period. Such an increase was also evident during the last 15 min before REM sleep onset. Results suggest that spectral analysis of the HRV provides additional information of the ultradian rhythmic behavior of the autonomic nervous system function beyond the traditional cardiovascular measurements (mean heart rate, blood pressure, etc.). In contrast to these measurements, which generally show a continuously decreasing cardiovascular activity, as the night proceeds, the results of this study reveal a high sympathetic peak activity during the later REM sleep periods, which is comparable in magnitude to that found in the upright position in wakefulness. This activity may be associated to the well-known incidence peak of ischemic events in the early morning hours. PMID- 9333198 TI - Effects of an activity-correlated feeding regime on circadian locomotor activity rhythms in LEW/Ztm rats. AB - The aim of the present study was to examine the possible feedback effect of activity on the circadian system in rats. The animals were housed in running wheel cages equipped with food dispensers and were forced to run for their food during parts of the study. Overall level and free-running period tau of wheel running activity were recorded continuously to quantify the relationship between tau and the level of activity. Surprisingly, the activity-dependent feeding regime failed to increase the level of wheel-running activity and did not affect the activity pattern. Nevertheless, the period of wheel-running activity was shortened in 50% of the animals subjected to food dispensers and in 38% of the animals fed only once a day at irregular times, indicating that subtle environmental differences can affect the circadian pacemaker system without changing the level or the pattern of activity. In addition, 26% of the animals showed a shortening of tau after 3 weeks, i.e., before any change in the experimental setup. This shortening was probably caused by the access to the running wheel at the beginning of the experiment. The present results suggest that the effect on the circadian pacemaker common to both of these experimental manipulations is a change in the physiological or emotional state of the animal rather than an increase of the overall level of activity. PMID- 9333199 TI - Dehydroepiandrosterone decreases behavioral despair in high- but not low-anxiety rats. AB - Outbred Sprague-Dawley rats exhibit considerable heterogeneity within a population when evaluated for a variety of biologic functions, such as dietary fat intake, alcohol preference, and expression of anxiety. To understand the neuroendocrine basis for depression and anxiety, we routinely assess outbred rats for behavioral despair (Porsolt's test), anxiety (elevated plus-maze), and urinary excretion of a variety of hormones. In one such study, we observed a significant correlation (r2 = 0.337; n = 30; p < 0.01) between the level of anxiety and the degree of behavioral despair. Within the above population, two distinct subgroups emerged: one with high anxiety and the other with low anxiety. We next evaluated the effect of dehydroepiandrosterone (DHEA), an anxiolytic neurosteroid, on the despair response in the two groups of rats. Treatment of high-anxiety rats with DHEA significantly diminished behavioral despair. In contrast, DHEA did not affect behavioral despair in low-anxiety rats. In conclusion, the results presented here show DHEA to be effective as an antidespair agent in rats with both high anxiety and despair. PMID- 9333200 TI - The pineal affects life span in hamsters with heart disease. AB - Cardiomyopathic hamsters (CMH) develop heart disease early in life which leads to congestive heart failure and death as these hamsters age. We have previously shown that living in constant light or other non-24-h light-dark (LD) cycles can increase longevity in these hamsters, and the current experiment examined potential mechanisms for this effect. Thus, CMH were orchidectomized, pinealectomized, or given melatonin treatment and then placed on either 1:23 or 1:23.6 LD cycles. Orchidectomy had no effect on longevity in either LD cycle, but in 1:23.6 it did lead to death with a greater degree of heart failure. On the other hand, pinealectomy of 1:23 CMH led to changes in life span similar to those produced by placing the hamsters in 1:23.6. Moreover, melatonin implant treatment of CMH in 1:23.6 led to changes in life span that were similar to those caused by life in 1:23, at least over the first half of the survival curves. Thus, it appears that the pineal gland and melatonin may be involved in mediating the effects of non-24-h LD cycles, whether these effects are beneficial or detrimental. In addition, the testes and testosterone appear to have no role in mediating these effects. These data suggest that inhibition, rather than stimulation, of pineal function might be beneficial for those with congestive heart failure, but further experiments are necessary to clarify when during the disease process potential treatments might be helpful. PMID- 9333201 TI - Perceptual learning in olfaction: professional wine tasters versus controls. AB - By having professional wine tasters and controls perform olfactory tasks of absolute detection (1-butanol), discrimination (lemon and cloves), and identification (common household odors), the present two experiments studied (a) if perceptual odor learning takes place from odor experience acquired under nonlaboratory conditions, (b) if this learning generalizes to odors for which experience is limited, and (c) if generalized learning can be referred to increased general interest for odors that increases attention to odorous features. The results showed that whereas wine tasters were not better than controls on detection, they were superior to controls on discrimination and identification, the latter due to only a few odors. Ratings of experience with certain odors during professional evaluation suggest that generalized perceptual learning may take place in discrimination but not in identification. Wine tasters did not show more general interest for odorous features than did controls. The nonsuperiority in detection may be explained by the fact wine tasters have no professional experience of a detection task per se, implying that perceptual odor learning does not generalize from the olfactory tasks of discrimination and identification to detection. PMID- 9333202 TI - Prostaglandin F2alpha-induced nest-building in pseudopregnant pigs. I. Effects of environment on behaviour and cortisol secretion. AB - Pigs may be susceptible to stress when they are strongly motivated to nest-build in a space-restricted environment. This study aimed to explore whether nest building behaviour could be induced by prostaglandin F2alpha (PG) administration to pseudopregnant gilts and to determine whether induced behaviour and cortisol output differed between animals chronically placed in either farrowing crates or pens. Jugular vein catheters were placed on Day 39 of pseudopregnancy and blood samples collected daily from Day 40 to Day 48. On Day 42, gilts were either restricted to farrowing crates 1.6 x 0.6 m with no straw (C: n = 11) or left in pens 2.8 x 1.74 m with straw (P: n = 11). On Day 47, blood and behaviour sampling was from 90 min pre-PG (Dinoprost; Lutalyse, Upjohn, Kalamazoo, MI) to 6 h post PG. PG injection successfully induced nest-building behaviour in P gilts within 15 min of injection. Penned gilts engaged in more straw/floor-directed behaviour than C gilts (p < 0.01), whereas attempts in C gilts seemed partial or incomplete. Conversely, C gilts showed increased (p < 0.05) amounts of fixture directed behaviour, whereas P gilts did not post-PG. For both groups, cortisol increased significantly (p < 0.05) post-PG compared to pre-PG values. Cortisol concentrations in C gilts were significantly greater than in P gilts prior to and after PG (p < 0.05) on Day 47, whereas there were no significant differences in concentrations of cortisol between C and P on other days. These results demonstrate that PG can induce nest-building behaviour in the absence of foetal signals. Whereas the pseudopregnant gilt seemed to chronically adapt to the imposition of a farrowing crate, gilts subsequently attempting to nest-build showed increases in cortisol output when compared to their penned counterparts, suggesting that a stress was imposed by the space-restricted environment. PMID- 9333203 TI - Prostaglandin F2alpha-induced nest-building in pseudopregnant pigs. II. Space restriction stress does not influence secretion of oxytocin, prolactin, oestradiol or progesterone. AB - We have previously shown that prostaglandin F2alpha (PG) is capable of inducing nest-building behaviour in pseudopregnant gilts and established a protocol. This experiment examined which reproductive endocrine systems might mediate these behavioural responses, in the presence or absence of a space restriction stress. Pseudopregnancy was induced with 5 mg/day i.m. (intramuscular) injections of oestradiol valerate (OV) on Days 11-15 of the oestrous cycle, jugular vein catheters were placed on Day 39 of pseudopregnancy, and blood samples were collected daily from Day 40 to Day 48. On Day 42, gilts were either space restricted to farrowing crates 1.6 x 0.6 m (C: n = 11) or left in pens 2.8 x 1.74 m (P: n = 11). On Day 47, blood samples were collected from all animals every 15 min from 90 min prior to a single i.m. injection of 15 mg of prostaglandin F2alpha (PG: Lutalyse, Upjohn, Crowley, West Sussex) to 120 min post-PG and then hourly for 4 h and assayed for oxytocin, prolactin, progesterone, and oestradiol. Results showed that mean daily concentrations of prolactin and progesterone were significantly lower (p < 0.05 respectively) in C than P gilts from Day 42 to Day 46 of pseudopregnancy. There were no significant differences in mean daily concentrations of oxytocin and oestradiol between C and P gilts during this time. For both groups, oxytocin, prolactin, and progesterone concentrations increased significantly (p < 0.05) post-PG when compared to their respective pre-PG values. However, for both groups, oestradiol concentrations were unaffected by PG injection. The prostaglandin-induced increases in oxytocin, prolactin, and progesterone concentrations did not differ between groups. We conclude that coincident changes in oestradiol secretion does not influence nesting behaviour and that space restriction stress associated with nest-building does not influence secretion of oxytocin, prolactin, oestradiol, or progesterone. PMID- 9333204 TI - Acute stress in pregnant rats: effects on growth rate, learning, and memory capabilities of the offspring. AB - Growth rate of the offspring of female rats stressed by the presence of a cat at the 10th or the 19th gestational day was lower than that of controls whereas footshocks administered at the same periods did not significantly influence growth rate of the young. Whatever the nature of the stress and the time when it was administered to the mother, the death rate of the young rats was much greater than that in controls. When adult, the offspring of stressed mothers exhibited learning and memory impairments in a delayed alternation task as well as in passive avoidance conditioning. Alteration of these cognitive functions is interpreted in terms of subtle dysfunctions in the development of the nervous system through modifications of the hormonal components of the mothers, particularly eventual alterations of the nervous system biochemistry of the offspring. PMID- 9333205 TI - Meal-induced changes in hepatic glycogen of fasted rats. AB - Hepatic metabolism of glucose and other nutrients influences feeding behavior. The present study was conducted to confirm prandial decreases in hepatic glycogen concentrations following a short-term fast. Male Sprague-Dawley rats were fasted 6-12 h during the light phase before having access to chow for one or two 20-min meals at the beginning of the dark phase. Plasma glucose and insulin concentrations in hepatic and portal venous blood and hepatic glycogen concentrations prior to and at the end of each meal were compared. Glucose concentration in the hepatic vein was greater than that in the portal vein prior to the meals but not at the end of the meal. Insulin levels were higher in the portal vein than the hepatic vein pre- and postprandially. Hepatic glycogen concentrations increased after each meal in younger (2-month-old) rats but not older (6-month-old) rats. Fasting levels of hepatic glycogen were lower in the younger rats than the older rats; however, the increase in hepatic glycogen was not due to differences in baseline glycogen concentrations at the start of the meal. The reported prandial decreases in hepatic glycogen of fasted rats were not apparent in this study. Because of the difference between 2- and 6-month-old rats in periprandial hepatic glycogen metabolism after a short-term fast, the age of the animal needs to be considered if the dynamics of liver glycogen metabolism are to be incorporated into a model of food intake regulation. PMID- 9333206 TI - Circadian performance of suprachiasmatic nuclei (SCN)-lesioned antelope ground squirrels in a desert enclosure. AB - Circadian activity parameters of 53 white-tailed antelope ground squirrels, Ammospermophilus leucurus, were measured to determine the role of the suprachiasmatic nuclei (SCN) pacemaker in their health and survival. Wheel running activity was monitored in the laboratory with 44 individuals to document the presence of free-running circadian rhythms and ability to entrain to light dark cycles. Twenty-two individuals were returned to the desert site of origin, including 12 intact control animals and 10 animals whose circadian timing had been disrupted by SCN-lesioning. Time of activity was recorded continuously for 15 days in a large outdoor enclosure by a motion detector, a microchip transponder detector, and video surveillance. An unplanned nighttime attack by a feral cat resulted in the death of 60% of the SCN-lesioned animals and 29% of the control animals in the enclosure. Surface activity of SCN-lesioned animals at the food cache occurred both in daytime and at night, ranging from 16.0% nighttime activity for one partially lesioned individual to 52.1% for one completely lesioned animal. Controls were strongly day-active, with nighttime surface trips constituting only 0-1.3% of all excursions to the cache. Nine wild free-ranging individuals, including one with a radiotransmitter collar, were exclusively day active. One of the functions of the SCN in mammals may be to reduce activity of animals during times that are unfavorable for activity. PMID- 9333207 TI - A novel stereotaxic-like method for injecting into the lateral ventricles of small passerine birds. AB - Recently, there has been an increase in the interest in various aspects of the biology of small passerine birds. In order to apply many modern neurobiological techniques, it would be useful to be able to surgically inject hormones and drugs directly into passerine brains. We describe a novel and cost-effective stereotaxic-like technique for injecting substances into the lateral ventricles of small passerine birds. The technique is rapid, reducing the period of anesthesia, and reliable. In addition, using this technique, we injected [3H]cysteine (to label newly synthesized proteins) with small doses of colchicine. Colchicine decreased the content of newly synthesized protein in the median eminence, thus suggesting that colchicine inhibited protein transport. The technique should prove useful in a number of neurobiologial applications. PMID- 9333208 TI - Behavioral and physiological sex differences observed in an animal model of fulminant hepatic encephalopathy in the rat. AB - Hepatic encephalopathy is characterized by a number of neuropsychiatric and motor disturbances observed in patients with liver dysfunction. The purpose of this study is to fully characterize behavioral and physiological sex differences in an animal model of fulminant hepatic encephalopathy (FHE). Male and female rats were administered thioacetamide (600 mg/kg) via i.p. (intraperitoneal) injection at Hours 0 and 24 and allowed to progress into the four stages of FHE. Male rats reached all four stages of FHE significantly earlier than female rats (p < 0.05). The performance of the male rats deteriorated more quickly (p < 0.05) than that of the females in all of the sensory and motor behavioral tests. Sex differences were observed in the liver enzymes of the FHE rats. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase were significantly greater (p < 0.05) in male rats in all four stages of FHE. Significant increases were also observed in the levels of direct and total bilirubin (p < 0.05). Neuronal damage was observed in the CA1 and CA2 regions of the hippocampus. In the CA1 region, male rats displayed greater pathological changes in Stages III and IV (p < 0.05) than female rats. The damage in the CA2 region was only observed in Stage IV male rats. Our data indicate that observable behavioral and physiological sex differences occur in thioacetamide-induced FHE in the rat. PMID- 9333210 TI - Detection of very complex taste mixtures: generous integration across constituent compounds. AB - Mixtures of compounds can often be tasted even when all of their components are too weak to be tasted separately. Such mixtures are said to be integrative. Integration was demonstrated by mixing compounds in concentrations proportional to their separate detection thresholds and then measuring the detection threshold of the mixture as a whole by forced choice with plain water. Mixtures of 3, 6, 12, and 24 compounds were thus evaluated. With earlier data on two-, three-, and four-component mixtures (Stevens, J. C. Detection of taste in mixture with other tastes: Issues of masking and aging. Chem. Senses 21:211-221; 1996.), the results show that the concentration of any constituent compound goes down in approximate proportion to the number of compounds with which it is in mixture. This nearly complete integration seems to describe mixtures of like-quality compounds, of unlike-quality compounds, and of both like- and unlike-quality compounds. Integrative mixtures of the sort studied here provide a model for the detection of the ultracomplex stimuli of everyday life, such as foods and drinking waters. Although the degree of integration may trail off slightly with mixtures of high complexity, the present result proffers no limit on the number of compounds that can be at least partially integrated. In principle, integration permits the detection of natural substances whose myriad components could all be far below threshold. The mechanism of taste integration is speculative, but the facts are congenial to the hypothesis of multiple parallel channels for the processing of intensity and quality. PMID- 9333209 TI - Contribution of gastric and postgastric feedback to satiation and satiety in women. AB - Two parallel preload studies were conducted to determine the relative contributions of inhibitory feedback from the stomach and intestine to satiation (meal termination) and postprandial satiety. In the Gastric Emptying Study, five normal-weight women each ingested an egg sandwich (307 kcal) (1) immediately after a tomato soup preload (120 kcal), (2) 20 min after a tomato soup preload, and (3) with no preload. There was 125 g more of soup in the stomach when subjects began ingesting the sandwich immediately compared to 20 min after the soup, and the emptying of the sandwich was delayed when it was ingested immediately but not 20 min after the soup. The lag times for emptying of the sandwich were 76.5 (69.1-82.4), 47.2 (20.1-67.7), and 42.4 (17.8-65.1) min for the three conditions, respectively, p < 0.05. In the Food Intake Study, 16 normal weight women ate significantly less (p < 0.01) in test meals offered immediately (978+/-246 kcal) and 20 min (1027+/-298 kcal) after the soup preload than in a test meal with no preload (1151+/-279 kcal). Despite the different amounts of soup in the stomach, subjects' test-meal intake in the two preload conditions was not significantly different. Subjects' fullness ratings following the preloads and the test meals were not different among the treatment conditions. The results suggest that feedback from neither the gastric nor the postgastric compartment is primary in determining meal size and postprandial satiety. Instead, signals from gastric and postgastric sources are combined to determine meal size and postprandial satiety. PMID- 9333211 TI - Drinking in sodium-depleted rats with bile duct, vena cava or portal vein obstruction. AB - The authors investigated whether a depletion of sodium with furosemide enhanced the water and 0.3 M NaCl intakes of rats with experimental cholestasis, portal hypertension or congestive heart failure. These were induced, respectively, by bile duct ligation (BDL), portal vein constriction or vena cava constriction. BDL alone increased daily saline intake. In BDL rats, but not in sham-ligated controls, experience with a prior depletion of sodium enhanced the 2-h saline intake and the retention of water and sodium after a subsequent depletion. Chronic cava constriction, but not portal constriction, enhanced sodium intake and retention after sodium depletion during a 2-h test and enhanced water intake overnight after the test. The results suggest that the ingestion of sodium by BDL and cava-constricted rats may share a common mechanism. PMID- 9333212 TI - The newborn rat ingests fluids through a surrogate nipple: a new technique for the study of early suckling behavior. AB - In this report we describe an apparatus and procedure that permits a newborn rat pup to ingest test fluids including milk through a surrogate nipple. The surrogate nipple represents a new testing situation for the experimental study of sensory and neurochemical controls of suckling behavior immediately after birth. PMID- 9333213 TI - Handling and isolation in three strains of rats affect open field, exploration, hoarding and predation. AB - Male albino (Al), brown hooded (Br) and black hooded (Bl) rats were raised in social isolation or in pairs, with or without systematic handling. At 90 and 180 days of age, the animals were individually tested for activity in an open field (Of), exploratory behavior in a complex environment, food hoarding (Hd) and insect predation (Pd). Multivariate analysis of the results showed significant influences of all three factors (strain, handling and social isolation) and interactions among them. Strain affected Of, Hd and Pd, with contrastingly high performances of Br in Of, Al in Hd and Bl in Pd. Handling increased Of and exploration scores in both test series. Isolation induced higher performances in all the four behaviors in the second test series. Accentuated and stable individual differences occurred in the performances off all the behaviors. The results emphasize the subtleness and complexity of the interplay of genetic and environmental influences and stress the independence of the regulatory processes of different behaviors. PMID- 9333214 TI - Daily rhythms in a complex operant: targeted percentile shaping of run lengths in rats. AB - Daily rhythms in response output and accuracy were examined when reinforcement for a complex operant was uncoupled from accuracy of performance. Rats housed in operant conditioning chambers earned their daily ration of food under a targeted percentile procedure for responding on two levers. The targeted pattern was a series of consecutive responses on the left lever (a "run"), followed by a single response on the right lever. The targeted run length was either "O" (i.e., undefined, under the nondifferential baseline), 6, 12 or 24. Under baseline, a random third of all trials ended in pellet delivery; under the percentile conditions, trials with runs closer to the target than two-thirds of the runs from the most recent 24 trials ended in pellet delivery. This contingency shaped run lengths while ensuring that approximately one-third of all trials produced pellets. Responding tracked the target value well, with mean obtained run lengths equal to 90% of the target or better. Daily rhythms were clearly evident in measures of overall output, with subjects responding primarily 3-7 h into the dark period. The only substantial light-period responding observed in all subjects occurred during the 2 h after noon, when the chambers were serviced. No systematic variation in this pattern was observed as a function of target. Run length was much less variable across the daily cycle than was response output, with only a suggestion under the longest target that response accuracy was lower during periods removed from the period of peak activity. PMID- 9333215 TI - Participation of the medial and anterior hypothalamus in the modulation of tonic immobility in guinea pigs. AB - Tonic immobility (TI) is an inhibitory behavioral response during which the animal presents profound physical inactivity and a relative lack of response to the environment. This response is induced in the laboratory by postural inversion of the animal and brief postural contention of its movements. In nature, the response occurs when there is physical contact between prey and predator. In this case, the physical inactivity of the prey may prevent the continuation of the attack. The neural substrate of this response is not well known and the objective of the present study was to investigate the effect of cholinergic stimulation of hypothalamic regions on TI modulation in guinea pigs (Cavia porcellus). Microinjection of carbachol (1.0 microg/0.2 microL) into the anterior hypothalamus promoted an increase in the duration of TI episodes. Microinjection of carbachol into the ventro- and dorsomedial hypothalamus, however, promoted a reduced duration of TI episodes. Pretreatment with atropine (0.5 microg/0.2 microL) showed that the action of carbachol is mediated by muscarinic receptors in the anterior and ventromedial hypothalamus but not in the dorsomedial hypothalamus. The results suggest that the hypothalamic regions may play different roles in the organization of defensive behavioral responses such as TI. PMID- 9333216 TI - Interleukin-1 (IL-1) receptor type I mediates anorexia but not adipsia induced by centrally administered IL-1beta. AB - IL-1beta induces anorexia and adipsia. Here, we report that intracerebroventricular (ICV) pretreatment with an antisense (but not sense) phosphothio-oligodeoxynucleotide to the IL-1 receptor type I (IL-1RI, 1.28 microg or 239 pmol twice daily for 3.5 days before IL-1beta plus antisense) inhibits the anorexia, but not the adipsia induced by the ICV administration of 2.0 ng IL 1beta/rat (a dose that yields estimated pathophysiological concentrations in the cerebrospinal fluid). The mean 2 h food intake decrease in response to IL-1beta was 5.6% (n = 10) in the antisense- and 43% in the sense (n = 9)-treated groups; the mean 2 h water intake decrease was 40% in the antisense- and 39% in the sense treated groups. The intraperitoneal administration of IL-1RI antisense, in doses equivalent to those administered centrally, had no effect on the anorexic effect induced by ICV administered IL-1beta; this indicates a direct action in the central nervous system. The results suggest that: i) IL-1RI is involved in the short-term anorexigenic, but not the adipsogenic effect induced by centrally administered IL-1beta; and ii) the approach presented using antisense strategies is applicable to study the molecular basis of IL-1 mediated behaviors. PMID- 9333217 TI - High lick rate is maintained throughout spontaneous liquid meals in freely feeding rats. AB - To investigate the microstructure of spontaneous meals in freely feeding rats, 16 adult male Sprague Dawley rats were housed individually in custom-designed lickometer cages and maintained on a milk diet. Licks were recorded over 23 h at millisecond accuracy via a computer-controlled lickometer. Analysis of lick data revealed an average of about 12 discrete meals/day occurring mainly during the dark phase. The most striking feature of both dark and light meals was the maintenance of a high initial rate of licking until an abrupt decline at the end of the meal. This pattern of licking is very different from the exponential decay of lick rate reported in scheduled test meals of palatable solutions. Thus, the microstructure of licking for meals is affected in an apparently fundamental way by whether a meal is scheduled or spontaneous, suggesting a basic difference in the underlying physiologic controls. PMID- 9333218 TI - Antiemetic inhibits conditioned taste aversion, but not the hypophagia induced by epinephrine. AB - It has been shown that relatively high doses of epinephrine (E) injected intraperitoneally (IP) produce hypophagia and conditioned taste aversion (CTA) in rats. We examined the possibility that E effects involve malaise. For this purpose, changes in saccharin preference induced by E injected IP (100 microg/kg) were determined after a previous administration of trimethobenzamide (TMB, 5 mg/kg), an antiemetic agent. E alone decreased saccharin preference by 54% (p < 0.01), but only by 16% (not significant) in the presence of TMB. In contrast, the injection of 75 or 100 microg/kg E reduced food intake by 50 and 85%, respectively (p < 0.01), regardless of previous injection of TMB. In conclusion, the results suggest that E-induced malaise is not the direct cause of the hypophagia it elicits. PMID- 9333219 TI - Flunarizine analgesia is mediated by mu-opioid receptors. AB - We evaluated the opioid antinociceptive mechanism of the calcium channel blocker flunarizine. Flunarizine produced a dose-dependent analgesia in the hot plate test which was antagonized by general (naloxone) and mu [beta-fualtrexamine (beta FNA)]-opioid antagonists, indicating a role for mu receptors in flunarizine analgesia. Naltrindole (delta1 antagonist) did not affect flunarizine analgesia. A fixed subthreshold dose of flunarizine augmented mu (morphine) analgesia and blocked delta1 [enkephalin, [D-Pen2,5] (DPDPE)] analgesia. Apparently, flunarizine has mu agonistic activity and delta1 antagonist or mixed agonistic antagonistic activity. PMID- 9333220 TI - Not the last word on the BSE crisis. PMID- 9333221 TI - US dispute over live AIDS vaccine trials. PMID- 9333222 TI - British guidelines set out standards for genetic tests. PMID- 9333223 TI - Nobel laureates face libel suits from 'water memory' researcher. PMID- 9333224 TI - NIH inquiry fails to find culprit of contamination. PMID- 9333225 TI - Cloning and bioethical thinking. PMID- 9333226 TI - European centres for disease control. PMID- 9333227 TI - The lost ape. PMID- 9333228 TI - Human BSE. PMID- 9333229 TI - Evolutionary biology. Pelvic problems for mammals. PMID- 9333230 TI - Molecular clocks. PERpetuating the PASt. PMID- 9333231 TI - Paleoanthropology. One skull does not a species make. PMID- 9333232 TI - The same prion strain causes vCJD and BSE. PMID- 9333233 TI - Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. AB - In blood coagulation, units of the protein fibrinogen pack together to form a fibrin clot, but a crystal structure for fibrinogen is needed to understand how this is achieved. The structure of a core fragment (fragment D) from human fibrinogen has now been determined to 2.9 A resolution. The 86K three-chained structure consists of a coiled-coil region and two homologous globular entitles oriented at approximately 130 degrees to each other. Additionally, the covalently bound dimer of fragment D, known as 'double-D', was isolated from human fibrin, crystallized in the presence of a Gly-Pro-Arg-Pro-amide peptide ligand, which simulates the donor polymerization site, and its structure solved by molecular replacement with the model of fragment D. PMID- 9333234 TI - Epipubic bones in eutherian mammals from the late Cretaceous of Mongolia. AB - An important transformation in the evolution of mammals was the loss of the epipubic bones. These are elements projecting anteriorly from the pelvic girdle into the abdominal region in a variety of Mesozoic mammals, related tritylodonts, marsupials and monotremes but not in living eutherian (placental) mammals. Here we describe a new eutherian from the Late Cretaceous period of Mongolia, and report the first record of epipubic bones in two distinct eutherian lineages. The presence of epipubic bones and other primitive features suggests that these groups occupy a basal position in the Eutheria. It has been argued that the epipubic bones support the pouch in living mammals, but epipubic bones have since been related to locomotion and suspension of the litter mass of several attached, lactating offspring. The loss of the epipubic bones in eutherians can be related to the evolution of prolonged gestation, which would not require prolonged external attachment of altricial young. Thus the occurrence of epipubic bones in two Cretaceous eutherians suggests that the dramatic modifications connected with typical placental reproduction may have been later events in the evolution of the Eutheria. PMID- 9333235 TI - The origin of the dog-like borhyaenoid marsupials of South America. AB - Dog-like marsupials (superfamily Borhyaenoidea) were the largest predacious mammals during the Tertiary period in South America. They are critical to our understanding of marsupial origin, phylogeny and palaeobiogeography because they have been related to various marsupial lineages of several continents: didelphoids (mainly New World, but also Europe, Asia and Africa), pediomyid, stagodontids (North America), dasyuroids (Australia) and deltatheroidans (predominantly Asian). These relationships, based mainly on dental morphology, have been discussed and rejected several times. Here we report the discovery of exceptionally well preserved skulls and skeletons, referrable to the didelphoid Andinodelphys, which shed new light on the phylogenetic and palaeobiogeographic origin of dog-like marsupials. The skulls of Mayulestes (boryhyaenoid), Andinodelphys and Pucadelphys (didelphoids) from the early Palaeocene epoch of Bolivia are the oldest known for American marsupials. Comparison of their basicranial anatomy suggests that dog-like marsupials are closely related to an early didelphimorphian radiation in South America, rather than to Asiatic (deltatheroidan), North American (stagodontid), or Australian (dasyuroid) lineages. PMID- 9333236 TI - The first skull of Australopithecus boisei. AB - Australopithecus boisei was first described from a cranium recovered in 1959 from Olduvai Gorge, Tanzania. This and subsequent finds, mostly from Kenya's Turkana basin, resulted in its characterization as a specialized Australopithecus species with a hyper-robust masticatory apparatus. A distinct A. boisei facial morphology has been emphasized to differentiate robust Australopithecus lineages from East and South Africa. A preference for closed and/or wet habitats has been hypothesized. Here we report some new A. boisei specimens, including the taxon's first cranium and associated mandible, from Konso, Ethiopia. These fossils extend the known geographical range of A. boisei. They provide clear evidence for the coexistence of A. boisei and Homo erectus within a predominantly dry grassland environment. The A. boisei specimens from Konso demonstrate considerable morphological variation within the species. The unexpected combination of cranial and facial features of this skull cautions against the excessive taxonomic splitting of early hominids based on morphological detail documented in small and/or geographically restricted samples. PMID- 9333237 TI - Decrease of cardiac chaos in congestive heart failure. AB - The electrical properties of the mammalian heart undergo many complex transitions in normal and diseased states. It has been proposed that the normal heartbeat may display complex nonlinear dynamics, including deterministic chaos, and that such cardiac chaos may be a useful physiological marker for the diagnosis and management of certain heart trouble. However, it is not clear whether the heartbeat series of healthy and diseased hearts are chaotic or stochastic, or whether cardiac chaos represents normal or abnormal behaviour. Here we have used a highly sensitive technique, which is robust to random noise, to detect chaos. We analysed the electrocardiograms from a group of healthy subjects and those with severe congestive heart failure (CHF), a clinical condition associated with a high risk of sudden death. The short-term variations of beat-to-beat interval exhibited strongly and consistently chaotic behaviour in all healthy subjects, but were frequently interrupted by periods of seemingly non-chaotic fluctuations in patients with CHF. Chaotic dynamics in the CHF data, even when discernible, exhibited a high degree of random variability over time, suggesting a weaker form of chaos. These findings suggest that cardiac chaos is prevalent in healthy heart, and a decrease in such chaos may be indicative of CHF. PMID- 9333238 TI - A specific neural substrate for perceiving facial expressions of disgust. AB - Recognition of facial expressions is critical to our appreciation of the social and physical environment, with separate emotions having distinct facial expressions. Perception of fearful facial expressions has been extensively studied, appearing to depend upon the amygdala. Disgust-literally 'bad taste'-is another important emotion, with a distinct evolutionary history, and is conveyed by a characteristic facial expression. We have used functional magnetic resonance imaging (fMRI) to examine the neural substrate for perceiving disgust expressions. Normal volunteers were presented with faces showing mild or strong disgust or fear. Cerebral activation in response to these stimuli was contrasted with that for neutral faces. Results for fear generally confirmed previous positron emission tomography findings of amygdala involvement. Both strong and mild expressions of disgust activated anterior insular cortex but not the amygdala; strong disgust also activated structures linked to a limbic cortico striatal-thalamic circuit. The anterior insula is known to be involved in responses to offensive tastes. The neural response to facial expressions of disgust in others is thus closely related to appraisal of distasteful stimuli. PMID- 9333239 TI - Transmissions to mice indicate that 'new variant' CJD is caused by the BSE agent. AB - There are many strains of the agents that cause transmissible spongiform encephalopathies (TSEs) or 'prion' diseases. These strains are distinguishable by their disease characteristics in experimentally infected animals, in particular the incubation periods and neuropathology they produce in panels of inbred mouse strains. We have shown that the strain of agent from cattle affected by bovine spongiform encephalopathy (BSE) produces a characteristic pattern of disease in mice that is retained after experimental passage through a variety of intermediate species. This BSE 'signature' has also been identified in transmissions to mice of TSEs of domestic cats and two exotic species of ruminant, providing the first direct evidence for the accidental spread of a TSE between species. Twenty cases of a clinically and pathologically atypical form of Creutzfeldt-Jakob disease (CJD), referred to as 'new variant' CJD (vCJD), have been recognized in unusually young people in the United Kingdom, and a further case has been reported in France. This has raised serious concerns that BSE may have spread to humans, putatively by dietary exposure. Here we report the interim results of transmissions of sporadic CJD and vCJD to mice. Our data provide strong evidence that the same agent strain is involved in both BSE and vCJD. PMID- 9333240 TI - AMPA receptor-mediated regulation of a Gi-protein in cortical neurons. AB - Excitatory synaptic transmission in the central nervous system is mediated primarily by the release of glutamate from presynaptic terminals onto postsynaptic channels gated by N-methyl-D-aspartate (NMDA) and alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors. The myriad intracellular responses arising from the activation of the NMDA and AMPA receptors have previously been attributed to the flow of Ca2+ and/or Na+ through these ion channels. Here we report that the binding of the agonist AMPA to its receptor can generate intracellular signals that are independent of Ca2+ and Na+ in rat cortical neurons. In the absence of intracellular Ca2+ and Na+, AMPA, but not NMDA, brought about changes in a guanine-nucleotide-binding protein (Galpha[il]) that inhibited pertussis toxin-mediated ADP-ribosylation of the protein in an in vitro assay. This effect was observed in intact neurons treated with AMPA as well as in isolated membranes exposed to AMPA, and was also found in MIN6 cells, which express functional AMPA receptors but have no metabotropic glutamate receptors. AMPA also inhibited forskolin-stimulated activity of adenylate cyclase in neurons, demonstrating that Gi proteins were activated. Moreover, both Gbetagamma blockage and co-precipitation experiments demonstrated that the modulation of the Gi protein arose from the association of Galpha(il) with the glutamate receptor-1 (GluR1) subunit. These results suggest that, as well as acting as an ion channel, the AMPA receptor can exhibit metabotropic activity. PMID- 9333241 TI - Prolonged photoresponses in transgenic mouse rods lacking arrestin. AB - Arrestins are soluble cytoplasmic proteins that bind to G-protein-coupled receptors, thus switching off activation of the G protein and terminating the signalling pathway that triggers the cellular response. Although visual arrestin has been shown to quench the catalytic activity of photoexcited, phosphorylated rhodopsin in a reconstituted system, its role in the intact rod cell remains unclear because phosphorylation alone reduces the catalytic activity of rhodopsin. Here we have recorded photocurrents of rods from transgenic mice in which one or both copies of the arrestin gene were disrupted. Photoresponses were unaffected when arrestin expression was halved, indicating that arrestin binding is not rate limiting for recovery of the rod photoresponse, as it is in Drosophila. With arrestin absent, the flash response displayed a rapid partial recovery followed by a prolonged final phase. This behaviour indicates that an arrestin-independent mechanism initiates the quench of rhodopsin's catalytic activity and that arrestin completes the quench. The intensity dependence of the photoresponse in rods lacking arrestin further suggests that, although arrestin is required for normal signal termination, it does not participate directly in light adaptation. PMID- 9333242 TI - The exocytotic event in chromaffin cells revealed by patch amperometry. AB - In mast cells and granulocytes, exocytosis starts with the formation of a fusion pore. It has been suggested that neurotransmitters may be released through such a narrow pore without full fusion. However, owing to the small size of the secretory vesicles containing neurotransmitter, the properties of the fusion pore formed during Ca2+-dependent exocytosis and its role in transmitter release are still unknown. Here we investigate exocytosis of individual chromaffin granules by using cell-attached capacitance measurements combined with electrochemical detection of catecholamines, achieved by inserting a carbon-fibre electrode into the patch pipette. This allows the simultaneous determination of the opening of individual fusion pores and of the kinetics of catecholamine release from the same vesicle. We found that the fusion-pore diameter stays at <3 nm for a variable period, which can last for several seconds, before it expands. Transmitter is released much faster through this pore than in mast cells, generating a 'foot' signals which precedes the amperometric spike. Occasionally, the narrow pore forms only transiently and does not expand, allowing complete transmitter release without full fusion of the vesicle with the plasma membrane. PMID- 9333243 TI - Circadian oscillation of a mammalian homologue of the Drosophila period gene. AB - Many biochemical, physiological and behavioural processes in organisms ranging from microorganisms to vertebrates exhibit circadian rhythms. In Drosophila, the gene period (per) is required for the circadian rhythms of locomotor activity and eclosion behaviour. Oscillation in the levels of per mRNA and Period (dPer) protein in the fly brain is thought to be responsible for the rhythmicity. However, no per homologues in animals other than insects have been identified. Here we identify the human and mouse genes (hPER and mPer, respectively) encoding PAS-domain (PAS, a dimerization domain present in Per, Amt and Sim)-containing polypeptides that are highly homologous to dPer. Besides this structural resemblance, mPer shows autonomous circadian oscillation in its expression in the suprachiasmaticnucleus, which is the primary circadian pacemaker in the mammalian brain. Clock, a mammalian clock gene encoding a PAS-containing polypeptide, has now been cloned: it is likely that the Per homologues dimerize with other molecule(s) such as Clock through PAS-PAS interaction in the circadian clock system. PMID- 9333244 TI - Head induction by simultaneous repression of Bmp and Wnt signalling in Xenopus. AB - The Spemann organizer of the amphibian embryo can be subdivided into two discrete activities, namely trunk organizer and head organizer. Several factors secreted from the organizer that are involved in trunk organization are thought to act by repressing Bmp signalling. With the exception of the secreted factor cerberus, little is known about head-organizer inducers. Here we show that co-expression of a dominant-negative Bmp receptor with inhibitors of the Wnt-signalling pathway in Xenopus leads to the induction of complete secondary axes, including a head. This induction does not require expression of the siamois marker of Nieuwkoop centre signalling, suggesting that cells are directly shifting to head-organizer fate. Furthermore, we find that cerberus is a potent inhibitor of Wnt signalling. Our results indicate that head-organizer activity results from the simultaneous repression of Bmp and Wnt signalling and they suggest a mechanism for region specific induction by the organizer. PMID- 9333245 TI - Protein memory through altered folding mediated by intramolecular chaperones. AB - The 77-residue propeptide of subtilisin acts as an intramolecular chaperone that organizes the correct folding of its own protease domain. Similar folding mechanisms are used by several prokaryotic and eukaryotic proteins, including prohormone-convertases. Here we show that the intramolecular chaperone of subtilisin facilitates folding by acting as a template for its protease domain, although it does not form part of that domain. Subtilisin E folded by an intramolecular chaperone with an Ile(-48)-to-Val mutation acquires an 'altered' enzymatically active conformation that differs from wild-type subtilisin E. Although both the altered and wild-type subtilisins have identical amino-acid sequences, as determined by amino-terminal sequencing and mass spectrometry, they bind their cognate intramolecular chaperones with 4.5-fold greater affinity than non-cognate intramolecular chaperones, when added in trans. The two subtilisins also have different secondary structures, thermostability and substrate specificities. Our results indicate that an identical polypeptide can fold into an altered conformation through a mutated intramolecular chaperone and maintains memory of the folding process. Such a phenomenon, which we term 'protein memory', may be important in investigations of protein folding. PMID- 9333246 TI - A piece of my mind. A 'normal' practice. PMID- 9333248 TI - Scientists revel in new research tool: an online index of cancer genes. PMID- 9333247 TI - Predisposition genetic testing for late-onset disorders in adults. A position paper of the National Society of Genetic Counselors. PMID- 9333249 TI - Center for genetic research on scleroderma. PMID- 9333250 TI - As discoveries unfold, a new urgency to bring genetic literacy to physicians. PMID- 9333251 TI - From the Centers for Disease Control and Prevention. Gonorrhea among men who have sex with men--selected sexually transmitted diseases clinics, 1993-1996. PMID- 9333252 TI - From the Centers for Disease Control and Prevention. Chlamydia screening practices of primary-care providers--Wake County, North Carolina, 1996. PMID- 9333253 TI - Social ties and susceptibility to the common cold. PMID- 9333254 TI - Social ties and susceptibility to the common cold. PMID- 9333255 TI - Prevention of bacterial endocarditis: American Heart Association recommendations. PMID- 9333256 TI - Prevention of bacterial endocarditis: American Heart Association recommendations. PMID- 9333257 TI - Prevention of bacterial endocarditis: American Heart Association recommendations. PMID- 9333258 TI - Issues regarding antiretroviral treatment for patients with HIV-1 infection. PMID- 9333259 TI - Issues regarding antiretroviral treatment for patients with HIV-1 infection. PMID- 9333260 TI - Issues regarding antiretroviral treatment for patients with HIV-1 infection. PMID- 9333261 TI - Issues regarding antiretroviral treatment for patients with HIV-1 infection. PMID- 9333262 TI - Prescribing protease inhibitors for the homeless. PMID- 9333263 TI - Treatment of verruca vulgaris with topical cidofovir. PMID- 9333264 TI - Complete genomic screen in late-onset familial Alzheimer disease. Evidence for a new locus on chromosome 12. AB - CONTEXT: Four genetic loci have been identified as contributing to Alzheimer disease (AD), including the amyloid precursor protein gene, the presenilin 1 gene, the presenilin 2 gene, and the apolipoprotein E gene, but do not account for all the genetic risk for AD. OBJECTIVE: To identify additional genetic risk factors for late-onset AD. DESIGN: A complete genomic screen was performed (N=280 markers). Critical values for chromosomal regional follow-up were a P value of .05 or less for affected relative pair analysis or sibpair analysis, a parametric lod score of 1.0 or greater, or both. Regional follow-up included analysis of additional markers and a second data set. SETTING: Clinic populations in the continental United States. PATIENTS: From a series of multiplex families affected with late-onset (> or =60 years) AD ascertained during the last 14 years (National Insititute of Neurological Disorders and Stroke-Alzheimer's Disease and Related Disorders Association diagnostic criteria) and for which DNA has been obtained, a subset of 16 families (135 total family members, 52 of whom were patients with AD) was used for the genomic screen. A second subset of 38 families (216 total family members, 89 of whom were patients with AD) was used for the follow-up analysis. MAIN OUTCOME MEASURES: Linkage analysis results generated using both genetic model-dependent (lod score) and model-independent methods. RESULTS: Fifteen chromosomal regions warranted initial follow-up. Follow-up analyses revealed 4 regions of continued interest on chromosomes 4, 6, 12, and 20, with the strongest results observed forchromosome 12. Peak 2-point affecteds only lod scores (n=54) were 1.3, 1.6, 2.7, and 2.2 and affected relative pairs P values (n=54) were .04, .03, .14, and .04 for D12S373, D12S1057, D12S1042, and D12S390, respectively. Sibpair analysis (n=54) resulted in maximum lod scores (MLSs) of 1.5, 2.6, 3.2, and 2.3 for these markers, with a peak multipoint MLS of 3.5. A priori stratification by APOE genotype identified 27 families that had at least 1 member with AD whose genotype did not contain an APOE*4 allele. Analysis of these 27 families resulted in MLSs of 1.0, 2.4, 3.7, and 3.3 and a peak multipoint MLS of 3.9. CONCLUSIONS: A complete genomic screen in families affected with late-onset AD identified 4 regions of interest after follow-up. Chromosome 12 gave the strongest and most consistent results with a peak multipoint MLS of 3.5, suggesting that this region contains a new susceptibility gene for AD. Additional analyses are necessary to identify the chromosome 12 susceptibility gene for AD and to follow up the regions of interest on chromosomes 4, 6, and 20. PMID- 9333265 TI - BRCA1 sequence analysis in women at high risk for susceptibility mutations. Risk factor analysis and implications for genetic testing. AB - CONTEXT: A mutation in the BRCA1 gene may confer substantial risk for breast and/or ovarian cancer. However, knowledge regarding all possible mutations and the relationship between risk factors and mutations is incomplete. OBJECTIVES: To identify BRCA1 mutations and to determine factors that best predict presence of a deleterious BRCA1 mutation in patients with breast and/or ovarian cancer. DESIGN: A complete sequence analysis of the BRCA1 coding sequence and flanking intronic regions was performed in 798 women in a collaborative effort involving institutions from the United States, Italy, Germany, Finland, and Switzerland. PARTICIPANTS: Institutions selected 798 persons representing families (1 person for each family) thought to be at elevated a priori risk of BRCA1 mutation due to potential risk factors, such as multiple cases of breast cancer, early age of breast cancer diagnosis, and cases of ovarian cancer. No participant was from a family in which genetic markers showed linkage to the BRCA1 locus. MAJOR OUTCOME MEASURES: Sequence variants detected in this sample are presented along with analyses designed to determine predictive characteristics of those testing positive for BRCA1 mutations. RESULTS: In 102 women (12.8%), clearly deleterious mutations were detected. Fifty new genetic alterations were found including 24 deleterious mutations, 24 variants of unknown significance, and 2 rare polymorphisms. In a subset of 71 Ashkenazi Jewish women, only 2 distinct deleterious mutations were found: 185delAG in 17 cases and 5382insC in 7 cases. A bias in prior reports for mutations in exon 11 was revealed. Characteristics of a patient's specific diagnosis (unilateral or bilateral breast cancer, with or without ovarian cancer), early age at diagnosis, Ashkenazi Jewish ethnicity, and family history of cancer were positively associated with the probability of her carrying a deleterious BRCA1 mutation. CONCLUSIONS: Using logistic regression analysis, we provide a method for evaluating the probability of a woman's carrying a deleterious BRCA1 mutation for a wide range of cases, which can be an important tool for clinicians as they incorporate genetic susceptibility testing into their medical practice. PMID- 9333266 TI - Characteristics of prostate cancer in families potentially linked to the hereditary prostate cancer 1 (HPC1) locus. AB - CONTEXT: Approximately 9% of prostate cancer cases have been estimated to result from inheritance of mutated prostate cancer susceptibility genes. Few data exist as to whether there are clinical differences between prostate cancers that are inherited and those that occur in the general population. OBJECTIVE: To investigate phenotypic characteristics of families potentially linked to the hereditary prostate cancer 1 (HPC1) locus on chromosome 1q24-25. DESIGN: Retrospective case study in which clinical data were extracted from medical and pathological records. FAMILIES: A total of 74 North American families with hereditary prostate cancer. Prostate cancer cases from the National Cancer Data Base were used as a reference population for comparison. MAIN OUTCOME MEASURES: The families were divided into 2 groups: either potentially linked (33 families with 133 men with prostate cancer), and thus likely to be carrying an altered HPC1 gene, or potentially unlinked (41 families with 172 men with prostate cancer), on the basis of haplotype analysis in the region of HPC1. The age at diagnosis of prostate cancer, serum prostate-specific antigen levels, digital rectal examination status, stage, grade, primary treatment of prostate cancers, and occurrence of other cancers were compared between the groups. RESULTS: The mean age at diagnosis of prostate cancer for men in potentially linked families was significantly lower than for men in potentially unlinked families (63.7 vs 65.9 years, respectively, P=.01; mean age at diagnosis in the reference population was 71.6 years). Higher-grade cancers (grade 3) were more common in potentially linked families, and advanced-stage disease was found in 41% of the case patients in potentially linked families compared with 31% in both the potentially unlinked families and the reference groups (P=.03 for the latter comparison). In the other clinical parameters, we found no significant differences between the groups. A modest excess of breast cancer and colon cancer was found in potentially linked families in comparison with potentially unlinked families, but this difference was not statistically significant. CONCLUSIONS: Families that provide evidence for segregation of an altered HPC1 gene are characterized by multiple cases of prostate cancer that, in most respects, are indistinguishable from nonhereditary cases. However, 3 characteristics were observed: younger age at diagnosis, higher-grade tumors, and more advanced-stage disease. Our study shows that a significant fraction of hereditary prostate cancers are diagnosed in advanced stages, emphasizing the clinical importance of early detection in men potentially carrying prostate cancer susceptibility genes. These findings support the current recommendations to screen men with a positive family history of prostate cancer beginning at age 40 years. PMID- 9333267 TI - Chromosome 19 single-locus and multilocus haplotype associations with multiple sclerosis. Evidence of a new susceptibility locus in Caucasian and Chinese patients. AB - CONTEXT: Susceptibility to multiple sclerosis (MS) involves a genetically complex autoimmune component. However, except for genes in the HLA system, specific susceptibility loci are unknown or unconfirmed. OBJECTIVE: To investigate several loci spanning 3 candidate regions for a role in multiple sclerosis (MS) susceptibility in 2 ethnic groups using both single-locus and haplotype analyses. The 3 regions include HLA on chromosome 6p21.3, APOE on chromosome 19ql 3.2, and MBP(myelin basic protein) on chromosome 18q23. DESIGN: Case-control association testing. SUBJECTS: A total of 120 Caucasian patients with MS and 107 unrelated control individuals from California, and 32 patients and 32 unrelated control individuals from Beijing, China. All patients with MS were diagnosed as having clinically definite disease according to published criteria. MAIN OUTCOME MEASURES: Chi2 Testing of loci and individual alleles and haplotypes. Haplotype frequencies were estimated with standard maximum likelihood methods. RESULTS: The HLA effect is due to the class II DR2 haplotype, DRB1*1501-DQA1*0102-DRB1*0602; contributions to MS susceptibility from additional DRB1-DQB1 alleles or other HLA region loci were not observed. Variation within the MBP locus on chromosome 18q23 showed no effect in MS. The distribution of haplotypes from 5 loci within the chromosome 19q13.2 region, including D19S178, D19S574, APOE, APOC2, and D19S219, differed between patient and control samples. D19S574 showed a significant effect (P=.015) in Caucasian patients with MS due to the increased frequency of a single allele (P=.002). The APOE variation, prominent in other neurological diseases, showed no influence on MS susceptibility, despite its location within the chromosome 19q13.2 region. Interaction effects between DR2 and chromosome 19q13.2 or MBP in MS susceptibility were not apparent. CONCLUSIONS: The significant chromosome 19q13.2 single-locus and multilocus haplotype associations with MS in Caucasian and Chinese patient samples indicate an effect from a nearby disease susceptibility locus. These initial observations are an encouraging step toward the description of non-HLA genetic susceptibility to MS. PMID- 9333268 TI - Cancer incidence after retinoblastoma. Radiation dose and sarcoma risk. AB - CONTEXT: There is a substantial risk of a second cancer for persons with hereditary retinoblastoma, which is enhanced by radiotherapy. OBJECTIVE: To examine long-term risk of new primary cancers in survivors of childhood retinoblastoma and quantify the role of radiotherapy in sarcoma development. DESIGN: Cohort incidence study of patients with retinoblastoma followed for a median of 20 years, and nested case-control study of a radiation dose-response relationship for bone and soft tissue sarcomas. SETTING/PARTICIPANTS: A total of 1604 patients with retinoblastoma who survived at least 1 year after diagnosis, identified from hospital records in Massachusetts and New York during 1914 to 1984. RESULTS: Incidence of subsequent cancers was statistically significantly elevated only in the 961 patients with hereditary retinoblastoma, in whom 190 cancers were diagnosed, vs 6.3 expected in the general population (relative risk [RR], 30 [95% confidence interval, 26-47]). Cumulative incidence (+/-SE) of a second cancer at 50 years after diagnosis was 51.0% (+/-6.2%) for hereditary retinoblastoma, and 5.0% (+/-3.0%) for nonhereditary retinoblastoma. All 114 sarcomas of diverse histologic types occurred in patients with hereditary retinoblastoma. For soft tissue sarcomas, the RRs showed a stepwise increase at all dose categories, and were statistically significant at 10 to 29.9 Gy and 30 to 59.9 Gy. A radiation risk for all sarcomas combined was evident at doses above 5 Gy, rising to 10.7-fold at doses of 60 Gy or greater (P<.05). CONCLUSIONS: Genetic predisposition has a substantial impact on risk of subsequent cancers in retinoblastoma patients, which is further increased by radiation treatment. A radiation dose-response relationship is demonstrated for all sarcomas and, for the first time in humans, for soft tissue sarcomas. Retinoblastoma patients should be examined for new cancers and followed into later life to determine whether their extraordinary cancer risk extends to common cancers of adulthood. PMID- 9333270 TI - Molecular diagnosis and carrier screening for beta thalassemia. AB - Thalassemias are common autosomal recessive disorders especially in populations of Mediterranean, Middle Eastern, and Far Eastern descent. Relatively high incidence is also observed in people of Asian Indian origin but the incidence is more limited in those of African descent. Beta Thalassemias are heterogeneous at the molecular level, with more than 150 different molecular defects identified to date. Despite this heterogeneity, each at-risk population has its own spectrum of common mutations, usually from 5 to 10, a finding that simplifies mutation analysis. Homozygosity for beta thalassemias usually results in transfusion dependent thalassemia major and, rarely, in mild non-transfusion-dependent conditions. Molecular diagnosis may be used to define genotypes associated with mild forms. Advances in carrier diagnosis using hematologic analysis followed by mutation analysis have made possible the population screening of women at childbearing age and prenatal diagnosis. This approach in combination with nondirective genetic counseling has resulted in a consistent decline of the birth of affected homozygotes in several Mediterranean at-risk populations, as well as knowledge of the risks of being a carrier. Molecular diagnosis of homozygotes and identification of carriers of beta thalassemia may lead to improved clinical management of patients with the disorder and prevention of the birth of affected homozygotes. PMID- 9333269 TI - Prenatal genetic carrier testing using triple disease screening. AB - CONTEXT: Rapid progress in gene discovery has dramatically increased diagnostic capabilities for carrier screening and prenatal testing for genetic diseases. However, simultaneous prenatal carrier screening for prevalent genetic disease has not been evaluated, and patient acceptance and attitudes toward this testing strategy remain undefined. OBJECTIVE: To evaluate an educational, counseling, and carrier testing program for 3 genetic disorders: Tay-Sachs disease (TSD), type 1 Gaucher disease (GD), and cystic fibrosis (CF) that differ in detectability, severity, and availability of therapy. DESIGN: Potential participants received education and genetic counseling, gave informed consent, chose screening tests, and completed pre-education and posteducation questionnaires that assessed knowledge, attitudes toward genetic testing, and disease testing preferences. SETTING: Medical genetics referral center. PATIENTS: Volunteer sample of 2824 Ashkenazi Jewish individuals enrolled as couples who were referred for TSD testing. INTERVENTION: Genetic counseling, education, and if chosen, genetic testing for any or all 3 disorders. MAIN OUTCOME MEASURE: Acceptance of screening for each of the 3 disorders. Secondary outcomes include attitudes toward genetic testing and reproductive considerations. RESULTS: Of the 2824 individuals tested for TSD, 97% and 95% also chose testing for CF and GD, respectively. The frequency of detected carriers was 1:21 for TSD, 1 :25 for CF, and 1:18 for GD. Twenty-one carriercoupleswere identified, counseled, and all postconception couples opted for prenatal diagnosis. Pre-education and posteducation questionnaires revealed that patients initially knew little about the diseases, but acquired disease information and increased knowledge of genetic concepts. Education and genetic counseling increased understanding and retention of genetic concepts and disease-related information, and minimized test-related anxiety. Although individuals sought screening for all 3 diseases, reproductive attitudes and decisions varied directly with disease severity and treatability. CONCLUSIONS: These findings emphasize the importance of genetic counseling for prenatal carrier testing and may improve understanding, acceptance, and informed decision making for prenatal carrier screening for multiple genetic diseases. PMID- 9333271 TI - Genetic testing in hereditary colorectal cancer. PMID- 9333272 TI - Molecular neurogenetics: the genome is settling the issue. PMID- 9333273 TI - Family history and genetic risk factors: forward to the future. PMID- 9333274 TI - Preparing health professionals for the genetic revolution. PMID- 9333275 TI - [Reconstruction of the nasal tip]. PMID- 9333276 TI - [Endocranial complications of acute and chronic otitis media in children and adolescents]. AB - BACKGROUND: After introduction of antibiotics endocranial complications in otitis media are less common, but there are still diagnostic and therapeutic problems. PATIENTS: 124 patients (5 months-16 years) underwent mastoidectomy. The number, type and therapy of otitic complications is presented. RESULTS: Nine percent of the patients showed up with endocranial complications. Besides lateral sinus thrombosis (five patients), meningitis (five patients) and epidural abscess (five patients), one cerebral and one cerebellar abscess was found. Four patients developed more than one complication. THERAPY: A complete mastoidectomy was performed with exploration of the dura and punction of the lateral sinus. In several cases these procedures gave the first diagnostic hint for endocranial complications. CONCLUSION: The occurrence of endocranial complications in the era of antibiotics should not be underestimated. Cholesteatoma is still a main risk factor for severe complications. If there is any doubt, further imaging studies (CT, MRI) are necessary. Diagnosis and therapy makes a close cooperation between neurosurgeon and ear surgeon and other specialties mandatory. PMID- 9333277 TI - [Methodological aspects of observations of rapid tympanic membrane movements due to changes in static pressure (Valsalva maneuver) using digital high speech video recordings]. AB - BACKGROUND: CCD-high speed camera systems are used to record high frequency videoclips. Introducing this technology in the analysis of eardrum motions, fast motions of the eardrum can be taped using an endoscope. METHODS: Records of the eardrum were made under specific static pressure conditions during the Valsalva maneuver. Initial experience with the help of a Kodak image (I 3) Ektapro 1000 Motion Analyzer allowed only conventional video signal processing and storage. A new data interface makes it possible to save and process the complete videoclip digitally. RESULTS: Using an image processing workstation and special mathematical algorithms, three-dimensional computer animations of rapid eardrum motions on static pressure can be performed. The present study describes different animations of fast motion eardrum movements. CONCLUSIONS: Digital high speed video imaging is a suitable method to describe rapid eardrum motions on static pressure (Valsalva maneuver). It is possible to visualize the motion of the eardrum over time on the fast Valsalva movement at high resolution. PMID- 9333278 TI - [Cochlear implant in patients with residual hearing]. AB - INTRODUCTIONS: Cochlear implants are used for the rehabilitation of bilaterally deaf patients. Due to the improvements in speech processing they might be also useful for patients with residual hearing and some speech understanding. METHODS: Pre- and postoperative speech understanding scores in 26 patients receiving implants were evaluated in a retrospective study. RESULTS: The preoperative pure tone threshold in the implanted ear was between 80 and 115 dB in the frequency range of 500 to 2000 Hz. On the contralateral side the mean threshold was 10 dB better. The mean score for the number test was 11.3%, for the monosyllables below 5%. Postoperatively speech understanding improved significantly up to 97% for numbers and 48% for monosyllables (tested with the Freiburger Speech Test). Cochlear implantation also benefits patients with residual hearing and some speech understanding with hearing aids under optimum conditions. The speech understanding scores must be below certain limits. General selection criteria cannot yet be specified. The individual decision must be based upon several criteria, especially the speech understanding scores in quiet and noise under optimum conditions. A prospective study is needed to develop generally applicable criteria. PMID- 9333279 TI - [Facial wrinkles--ultrapulsed CO2 laser: alternative or supplement to surgical face lift?]. AB - BACKGROUND: For many years, the treatment of facial wrinkles has been performed exclusively by dermabrasion, chemical peeling, or surgical face lifting. However, the recently introduced carbon dioxide lasers which emit ultrashort coherent light beams enable the cosmetic surgeon to ablate superficial epidemic layers with absent or very limited side effects. The purpose of this paper is to compare laser skin resurfacing with classical face lifting and discuss the potentials and limitations of each method. METHODS AND PATIENTS: Three patients suffering from facial wrinkles on photoaged skin were treated with the ultrapulsed CO2 laser (UltraPulse 5000 C; wavelength 10,600 nm, pulse duration 0.6 to 0.9 ms, maximum pulse energy 500 mJ). This laser guarantees vaporization of very thin superficial skin layers without scarring and with minimizing lateral thermal injury due to extreme short pulse duration. A special handpiece (CPG) permits an exact approach and a bloodless ablation of relatively large areas of facial skin. The fourth patient underwent a surgical face lift due to the depth of wrinkles. RESULTS: Excellent cosmetic results were achieved in all three patients with superficial wrinkles who were treated by laser skin resurfacing. When treating deeper wrinkles, e.g., glabella or nasolabial fold, the surgical face lift is the preferred method. CONCLUSION: Ultrapulsed CO2 laser treatment expands the therapeutic options for superficial facial wrinkles, especially for perioral, periorbital, forehead, and cheeks wrinkles. It proves to be a safe and effective method with very limited if any side effects. Nevertheless, deeper wrinkles are still a domain of the classical face lift. The combination of both methods may improve the overall outcome in the future. PMID- 9333280 TI - [Fine needle capillary cytology versus fine needle aspiration cytology--a comparison of quality between puncture techniques in the ENT area]. AB - BACKGROUND: Needle aspiration is currently a widely used technique in the diagnosis of unclear lesions in the head and neck region. We present a modified technique of fine needle biopsy in ENT, "fine needle capillary technique". The basics of this technique were developed by Zajdela and coworkers (1987) as a cytological method of fine needle biopsy in benign and malignant mammary tumors. Fine needle capillary technique does not require aspiration of cell samples via negative pressure created by a syringe. A thin 25 G needle (outer diameter 0.50 mm, length 25 mm) is introduced into the lesion with one hand. The cells are detached by the cutting edge of the needle and are conducted into the lumen by capillary force. The needle is removed and the cellular material is expelled onto a glass slide, spread, and immediately fixed. METHOD: In a series of 166 patients with unclear lesions in the head and neck region, we compared the fine needle capillary technique with the classic fine needle aspiration technique in each patient. Regarding quality and assessment of the cytological smear the fine needle capillary technique proved clearly superior in most of the cases. Lymph nodes, tumors of the salivary glands, thyroid glands, branchiogenic cysts, one atheroma, one lipoma, and one skin metastasis of a squamous cell carcinoma were punctured. RESULTS: In our study fine needle capillary technique showed a very good quality of the cytological smear in 24.7% of all cases, while fine needle aspiration technique reached 12.1% only. A good quality was obtained in 51.2% with fine needle capillary technique and in 51.8% with fine needle aspiration technique, poor quality in 24.1% with fine needle capillary technique and in 36.1% with fine needle aspiration technique. Nondiagnostic cytology was obtained in 21.7% with fine needle capillary technique and in 32.5% with fine needle aspiration technique. Both techniques together showed insufficient material in 10.8%. The quality of the cytological smear in each region was always better with fine needle capillary techniques than with fine needle aspiration technique except five punctures of the submandibular gland. Of 166 patients 113 (68.1%) underwent surgery, and a correlation of the cytologic report to the surgical specimen showed agreement in 95.7% with fine needle capillary technique and in 90.5% with fine needle aspiration technique. In 17.7% with fine needle capillary technique and in 25.7% with fine needle aspiration technique it was not possible to compare the cytological smear with the histological results because of poor quality of the cytological smear. In four cases (4.3%) with fine needle capillary technique the cytological diagnosis was wrong. With fine needle aspiration technique, a wrong diagnosis occurred eight cases (9.5%). CONCLUSIONS: Fine needle capillary technique offers several advantages. Without aspiration trauma to cells and tissues is reduced. Less blood in the samples results in higher quality of the cytological smear. These circumstances make it easier for the pathologist to comment the cytological findings. The handling of the needle is practiced with a wrist movement and not from the shoulder joint as in aspiration method using the Cameco syringe holder. This allows for a more sensitive puncture technique touching the lesion during sampling with the finger tips. The puncture causes less pain than the aspiration technique. Our results demonstrate that fine needle capillary technique is the better method of fine needle biopsy in the head and neck region. PMID- 9333281 TI - [Single suture closure and surgical staplers in neck interventions after previous radiotherapy]. AB - BACKGROUND: Impaired wound healing has to be expected more often in a tissue with diminished circulation. The technique of wound closure could have some influence on the healing result in such situations. METHODS: This prospective randomized study was carried out to evaluate whether staples or nylon sutures give better results regarding primary wound healing. Fifty patients who had previously been irradiated for head and neck tumors and underwent secondary surgery of the neck region were included in the study. RESULTS: Four of 26 wounds closed by conventional sutures evidenced delayed healing, while one of 24 wounds closed by stapler showed signs of impaired wound healing. This difference was not statistically significant. Further parameters (duration of operation, high age, high dosage of irradiation, long interval between irradiation and operation, previous chemotherapy, anemia, low body weight, relaxing incision, pharyngotomy) were also not associated with a higher incidence of delayed wound healing. CONCLUSIONS: Even in the irradiated cervical operation field no advantage for staples could be demonstrated in comparison with nylon sutures. PMID- 9333282 TI - [Toxic inner ear damage in topical treatment of psoriasis with salicylates]. AB - BACKGROUND: Whereas cochlear impairment after intravenous ingestions of salicylates is well known, reports of cochlear symptoms after topical application are quite rare. The extent of the ototoxic salicylate impact is increased by diabetes, renal insufficiency, and alcoholism. PATIENT: This study presents a case report of a female patient who suffered a repeated, symmetric, pancochlear, reversible inner ear impairment after two treatments with salicylate containing ointment for psoriasis. The correlation of the salicylate therapy with the observed inner ear lesions is obvious due to the close interval between these incidences and to the audiologic criteria typical for salicylate intoxication. CONCLUSION: Audiologic controls should be carried out during extended local application of salicylate containing ointment. PMID- 9333283 TI - [Chondroma of the petrous bone. A contribution to differential skull base tumor diagnosis]. AB - INTRODUCTION: Cartilaginous tumors of the mid-face and the skull base are rare. CASE REPORT: For the first time, a case report of a chondroma of the base of the ear in a 56-year-old woman is presented. In 1974 the patient developed a facial nerve paralysis while she was pregnant. Twenty-two years later the patient developed persistent headache and CT studies of the head were obtained, which showed an extensive tumorous lesion located at the base of the ear. A tumor was resected through an otoneurosurgical approach. The histological examination showed a chondroma. CONCLUSIONS: Even the rare diagnosis of a chondroma should be considered for a differential diagnosis of skull base tumors. PMID- 9333284 TI - [Unusual metastasis of hypernephroma in the maxillary sinus]. AB - BACKGROUND: Metastases of infraclavicular malignancies to the nose and paranasal sinuses are characterized by their untypical symptoms and late clinical manifestation. Metastases of renal carcinoma on paranasal cavities may be highly vasculared and can cause profuse bleeding. PATIENT: We report a rare case of a 60 year-old man who suffered from a large metastatic hypernephroma to the right maxillary sinus. Intermittent epistaxis and pain were the first symptoms. The patient had undergone a tumor nephrectomy six months previously. The endonasal biopsy of the vascular tumor caused abundant bleeding which required surgical treatment. RESULTS: The solitary metastasis was resected by a maxillectomy with closure of the large cavity with a microvascular latissimus dorsi flap. Additionally, a conservative treatment of interleukin-2 and alpha-interferon was administered. CONCLUSIONS: In case of a solitary metastasis of a renal carcinoma, a radical resection should be considered to improve the prognosis. The possibility of extensive perfusion should be taken into account when taking a biopsy of the lesion in order to be prepared for managing bleeding. PMID- 9333285 TI - [Postoperative pain therapy using on-demand analgesia after external ear reconstruction with rib cartilage harvesting]. AB - BACKGROUND: The individual experience of postoperative pain values considerably. It is the patient who can best evaluate the intensity of pain and the effect of a given analgesic substance. Patient-controlled analgesia (PCA) enables the patient to self-administer an i.v. analgesic bolus by pressing a button of an electronic pump device within a previously set range. If needed. The resection of rib cartilage to be used for ear reconstructions is associated with intense postoperative pain. This could result in unfavorable stress responses such as impaired cough and clearance of lung secretions carrying a greater risk of bronchitis or pneumonia, prolonged mobilization, and complications of wound healing. Therefore, individually adapted pain therapy is of great importance and technically feasible with modern PCA devices. METHOD: Informed and cooperative children aged five years and older and adults without severe health risks are suitable for PCA. Management, handling, monitoring and documentation of the PCA device are discussed. A 24-hour counselling service is maintained for occurring problems. RESULTS: Almost all patients expressed their great satisfaction with the means of pain therapy. Few patients were apprehensive. The initial scepticism of the involved personnel concerning opioid abuse, over-dosage, or technical problems gave way for a support of the PCA once the great satisfaction of the patients in absence of side effects was witnessed. No patient had to be antagonized for respiratory depression. Nausea and vomiting were rare events. CONCLUSION: Patient-controlled analgesia proved to be an excellent answer to postoperative pain after resections of rib cartilage. PMID- 9333286 TI - [The interesting case no. 3. Cholesteatoma with brain abscess and irritant meningitis]. PMID- 9333287 TI - [Salivary glands and the organism--interrelations and correlating reactions]. AB - BACKGROUND: Functional disturbances and diseases of the salivary glands are not only conditioned by local factors, but are also based on the manifold connections of the salivary glands with the remaining organism. The salivary glands represent consequently a "mirror of diseases" and a "diagnostic fluid". METHODS: A comparing analysis of the data of the Salivary Gland Register Hamburg (more than 18,000 cases) with the details of the literature was performed. RESULTS: Interrelationships and correlations of the salivary glands exist especially in diseases of the immune system, in viral infections, in metabolic diseases and in diseases of the hormone or nerve system. The occurrence and concentration of a great number of substances identified in the saliva represent a monitoring system with conclusions to functional or morphological alterations of other organs. CONCLUSION: The salivary glands as an integral component of the oral cavity are connected consequently in a manifold manner with the remaining organism by interrelationships and correlations and represent thus a "mirror of diseases". PMID- 9333288 TI - [Reconstruction of the side of the nose]. PMID- 9333289 TI - [Physiologic approach to promoting circulation in acute labyrinth ischemia. A new treatment concept]. AB - BACKGROUND: One of the main factors of sudden hearing impairment, acute vestibular disturbance, and acute tinnitus is generally thought to be an acute labyrinthine ischemia of varying degrees. However, the scientific basis for this assumption has not yet been proven, and there is a great variety of treatment modalities. Recent circulation research and fundamental physiological considerations led to the development of a new concept of treatment of these diseases. METHODS: In order to enhance the labyrinthine blood circulation, physiological and physical methods were tested in 42 patients, especially with regard to patient compliance and in addition to conventional therapy. The methods tested included active circulation training. Finnish bath observing certain precautions, sequentially increased temperature bathing, massage, and fango therapy of the neck. RESULTS: The compliance of the patients to the new concept was good, in some cases even enthusiastic, either to the "total body" methods or to the local neck treatment. CONCLUSION: The principle of active and/ or passive stimulation of blood circulation in acute labyrinthine ischemia has been well accepted by all patients. A study including functional results is in preparation. PMID- 9333290 TI - [Significance of vascular endothelium for specific functional behavior of nasal mucosa]. AB - BACKGROUND: During the last decade the morphological structure of nasal blood vessels was understood as a functional basis for different physiological and immunological reactions. RESULTS: Under normal conditions the vascular endothelium is involved in many activities including hemostasis, defense reactions, and angiogenesis during repair and pathological events. Most of the required components are synthesized by the endothelium itself, including extracellular matrix components, coagulation reactants, and growth factors. The complex vasculature of the human nasal mucosa plays an important role in the protection of the respiratory tract by generation of nasal fluid under normal and pathologic conditions. Increased plasma exudation in inflammatory processes including increased plasma protein concentrations binds cytokines, mediators, and cell release products and transports them to the respiratory lumen. During inflammation, endothelial cells stimulated by cytokines like TNF (tumor necrosis factor) regulate the recruitment of leukocytes by expression of certain surface proteins, called adhesion molecules. They also secrete chemokines like IL-8 (interleukin-8) and MCP (monocyte chemotactic protein) to increase the affinity of adhesion molecules, spread the leukocyte form to a flat, migrating one, and stimulate cell locomotion. Last but not least, endothelial cells may also act as antigen presenting cells (APC), although antigen presentation by endothelium is not responsible for the generalized homing of T-cells once the response is working. CONCLUSION: The new scientific results on the functional behaviour of vascular endothelium have opened new perspectives for our understanding of nasal blood vessels; it might also stimulate and enable us to search for new therapeutical concepts based on these findings. PMID- 9333291 TI - [Electron microscopy studies of vascular innervation of nasal mucosa in the human]. AB - BACKGROUND: The distribution of neural structures in human nasal vasculature is still not completely understood. To date, immunocytochemical and histochemical studies of the innervation pattern and neurotransmitter distribution have only been performed using light microscopy. The aim of this study was to verify these results by electron microscopy and to obtain new knowledge of the innervation of the individual vessel types. The target was to determine the role of the different vessels in the swelling mechanism in human nasal mucosa. MATERIAL AND METHODS: Specimens were prefixed in glutaraldehyde and postfixed in osmium tetroxide. After dehydration they were embedded in araldite. Ultrathin sections were cut, contrasted with uranylacetate and lead citrate, and studied under EM. RESULTS: Nasal vasculature is controlled by a dense innervation. Amyelinated longitudinal nerve bundles and smaller axon conglomerates are detected in the arterial adventitia. Capacitance vessels (veins) in general show few nerve structures, which, in contrast to arteries, are also located between the smooth muscle cells. Cushion veins, however, reveal a stronger innervation pattern especially in their muscular swelling. In contrast to smaller arterioles, no axons can de demonstrated in capillaries. CONCLUSION: The density of innervation in arteries and cushion veins seems to indicate that these vessel types are the central neural control point for the swelling mechanism in human nasal mucosa. Capillary function, however, does not seem to be directly influenced by the autonomic nervous system. PMID- 9333292 TI - [Receptor mediated gene transfer in squamous epithelial carcinoma of the head neck area]. AB - BACKGROUND: The "Adeno-Virus-Enhanced-Receptor-Mediated" (AVET)-System uses the receptor-mediated endocytosis route to introduce DNA into mammalian cells. By the help of a replication defective adenovirus with endosomolytic activity the coentry of receptor-bound DNA particles is faciliated. The transport to the nucleus followed by gene expression is multifold enhanced. METHODS: Two different permanent cell lines from head and neck squamous cell cancer were transfected with the reportergene for luciferase with either AVET or different liposoms in vitro. RESULTS: In comparison it becomes obvious, that AVET shows a multifold higher transfection rate in vitro than the liposomal formulations applied. CONCLUSIONS: AVET could be used for efficient transfection of epithelial cells, that are fairly reluctant to DNA transfer. Further trials in the mouse model must prove a possible application in vivo. PMID- 9333293 TI - [Detection of an "autocrine loop" between EGF receptor and TGF-alpha in head-neck carcinomas]. AB - BACKGROUND: In order to find a prognostic marker for the course of disease in head and neck cancer, we hypothesized that patients with rapid disease progress would produce increased levels of transforming growth factor alpha (TGF-alpha) and its cell surface receptor, the epidermal growth factor receptor (EGFR). METHODS: Using molecular biological techniques, we examined the incidence of TGF alpha and EGFR expression in 43 patients with tumors of the head and neck. The expression data were correlated with the course of disease in four-year follow up. The tumors were classified into four groups according to the EGFR status: Group 1, no expression for EGFR (15 samples); group 2, expression level 10 for EGFR (18 samples); group 3, expression level 50 for EGFR (7 samples); and group 4, expression level 100 for EGFR (3 samples). RESULTS: An expression for the TGF alpha protein was only detected in group 4. There was a significant correlation with EGFR overexpression in group 4 and survival compared with group 3 (p < 0.01) and group 1 (p < 0.05). The mean survival for group 1 to 4 was 27, 33, 34, and 10 months, respectively. CONCLUSION: The analysis of all patients revealed that the patients with synchronous expression of EGFR and TGF-alpha had the poorest prognosis. Increased production of TGF-alpha and EGFR in tumors of the head and neck may serve both as a marker for tumor progression and as a target for preventive therapies. PMID- 9333294 TI - [Fibroblast growth factor (b-FGF) in serum and urine of patients with head-neck malignancies]. AB - BACKGROUND: The angiogenic peptide basic fibroblast growth factor (b-FGF) has been suggested to significantly promote angiogenesis; a central event for the growth and metastasis of solid tumors. Elevated levels of b-FGF in the serum and urine of patients with various types of cancer have been reported. No information exists about b-FGF levels in patients with primary head and neck cancer. The present study was performed to determine serum and urine levels of b-FGF in these patients. METHODS: b-FGF was quantified in the urine of 50 (62.5%) patients with head and neck cancer as well as 30 (37.5%) patients with diseases unrelated to cancer using an immunoassay with a detection limit of 1.0 pg/ml (FGF basic "Quantikine", DFB00. Biermann GmbH, Bad Nauheim, Germany). Urine was collected from patients before breakfast and centrifuged for 10 min at 1000 g. The supernatants were stored at -80 degrees C until the assay was performed. RESULTS: In the serum and urine of all patients, b-FGF was detectable by immunoassay. Cancer patients revealed significant increased serum b-FGF concentrations, in addition, advanced tumor size (T3/4) showed significant increased b-FGF levels in both serum and urine. There seems to be a correlation between serum b-FGF concentrations with degree of histologic differentiation. CONCLUSION: The results of this study demonstrate that b-FGF levels are elevated in serum and urine of patients with primary head and neck cancer. These findings suggest an involvement of b-FGF in the formation of solid tumors. The value of b-FGF as a non-invasive monitoring of treatment response in cancer therapy, the correlation with angiogenesis in histologic sections as well immunohistochemical detection are the aim of further investigations. PMID- 9333296 TI - [History of the tuning fork. III: On the way to quantitative pure-tone measurement. Pictures from the history of otorhinolaryngology, represented by instruments from the collection of the Ingolstadt German Medical History Museum]. AB - BACKGROUND: Weber's and Rinne's tuning-fork tests were for a long time considered unreliable, as they often seemed to yield inconsistent results. The sources of error were manifold and lay in the fields of physics, physiology, pathophysiology, and psychology. When the problems came to be understood, more sophisticated instruments and techniques were developed. TECHNICAL IMPROVEMENTS IN TUNING FORKS: The prongs of the tuning fork were fitted with clamps to deaden overtones when it was put into vibration (Politzer 1870). By shifting the clamps along the prongs the tone of the tuning fork could be varied in a range up to one octave (Konlg 1878). A knob of hom or metal was fixed to the end of the shaft to ensure a good coupling to the skull when testing bone conduction (Lucae 1886). A small hammer fixed to the shaft and driven by a spring would activate the tuning fork with reproducible strength (Lucae 1899). A wedge-shaped figure drawn on the lateral surface of the clamps would allow one to optically control the amplitude of vibration (Gradenigo 1899). METHODS FOR QUANTIFICATION OF MEASURING HEARING ACUITY: The time during which a patient hears the tuning fork after it has been struck as compared to that of a normal hearing subject was measured as parameter of hearing acutiy (v. Conta 1864). A number of tuning forks at intervals of one octave each were assembled in sets to cover the whole frequency range of hearing. The most sophisticated example of these sets was the Bezold-Edelmann continuous tone series (1894). It comprised ten tuning forks with sliding clamps, two pipes of the organ type, and a Galton whistle. With this instrumentation it was possible to test the whole range of hearing. GRAPHIC PRESENTATION OF QUANTITATIVE RESULTS OF HEARING TESTING: The results of testing the hearing via air conduction and bone conduction measured in duration and calculated as percentage of normal hearing were presented in charts (Hartmann 1885, Gradenigo 1893) which can be considered precursors of modern audiograms. The evolution of these instruments and methods is described in detail and illustrated by exhibits from the museum. PMID- 9333295 TI - [Oropharyngeal fat hernia in Madelung's neck as a rare cause of acute dysphagia]. AB - BACKGROUND: Multiple symmetric lipomatosis (MSL) is a rare disorder clinically characterized by the presence of nonencapsulated lipomatous tissue around the neck, shoulders, and upper part of the thorax. The entity is frequently associated with peripheral neuropathy, chronic hepatopathy, and alcoholism. Despite of the cosmetically aspect of the patient with a reduced range of motion of the head and neck, MSL is asymptomatic in most cases. CASE REPORT: A 61-year old man with Madelung's disease complaining of acute dysphagia is presented. Magnetic resonance imaging revealed an oropharyngeal fat prolapse originating from the lipomatous tissue at the neck site. A symmetric resection of the neck fat with removal of the fat prolapse resulted in a satisfying functional and cosmetically outcome. CONCLUSION: To prevent complications and cosmetically disfigurement, early surgical intervention is recommended. PMID- 9333297 TI - [Anatomic terminology and nomenclature for paranasal sinus surgery]. AB - A consensus on the preferred modern usage of potentially confusing or ambiguous terms in sinus anatomy and nomenclature is described. These terms are intended to provide clear communication among otorhinolaryngologists worldwide and serve as a basis for discussion among anatomists. Terminology is based on Latin nomenclature. An attempt has been made to reconcile or eliminate duplication, redundancy, and overlap in terminology that have arisen over the past century. A key concept is that the ethmoid complex is divided into anterior and posterior sections by the basal lamella of the middle turbinate. PMID- 9333298 TI - [The interesting case no. 4. Right-sided elongated styloid process]. PMID- 9333299 TI - [Latin American science form the clinical perspective]. PMID- 9333300 TI - [Fungi and biodiversity: international incentives]. AB - Interest in biodiversity has spawned international actions both political and scientific, many of which are relevant to mycologists. The political initiatives include the Convention on Biological Diversity and Biodiversity Action Plans, and the scientific: the Global Biodiversity Assessment, Species 2000, Systematics Agenda 2000 International, BioNet International, Diversitas, All Taxa Biodiversity Inventory, and Biodiversity Methods Manuals. Mycologists need to contribute to such programmes, maintain an enhance profile, and remain flexible to respond to new challenges. PMID- 9333301 TI - [Internet, a new source of information]. PMID- 9333302 TI - [A candle in the dark]. PMID- 9333303 TI - [Studies on mineral waters bottled in glass, plastic and brick containers. 2. Development of total microbial flora]. PMID- 9333304 TI - [Food poisoning: a survey on intervention methodology in 6 public health units of the Lombardy Region. 2. Sporadic forms of salmonellosis]. PMID- 9333306 TI - [Cesium-137 and Americium-141 levels in foodstuffs]. PMID- 9333305 TI - [Foods responsible for transmission of cholera: factors favoring survival of Vibrio and preventive measures]. PMID- 9333307 TI - [Dietary lead exposure]. PMID- 9333308 TI - [Vaccinations during childhood; knowledge, opinions and attitudes of a sample of Italian mothers]. PMID- 9333310 TI - [Consumption, attitudes and knowledge on compared alcoholic beverages among high school and university students]. PMID- 9333309 TI - [Survey on knowledge about AIDS among high school and university students]. PMID- 9333311 TI - [Studies on mineral waters bottled in glass, plastic and brick containers. 1. Evaluation of microbiological aspects]. PMID- 9333312 TI - Some questions on specialisation. PMID- 9333313 TI - A question of balance. PMID- 9333314 TI - [Low molecular weight heparin: the first 5 years]. PMID- 9333316 TI - [Clinical significance and predictive value of laboratory tests in thrombosis associated with antiphospolipid antibodies]. AB - Antiphospholipid antibodies are a wide ranging, heterogeneous family of autoantibodies, formerly believed to be directed to anionic phospholipids. Recent research, however, has confirmed that they are directed to plasma proteins bound to suitable (phospholipid) anionic surfaces. The most well-known and best characterized antigens are beta 2-glycoprotein I, recognized by anticardiolipin antibodies, and prothrombin, recognized by most lupus anticoagulants. Lupus anticoagulants are generally identified on the basis of their capacity to prolong the phospholipid-dependent coagulation tests. Two types of lupus anticoagulants, anticardiolipin-type A, and antiprothrombin antibodies, whose presence is associated with different coagulation profiles, have been identified. Anticardiolipin-type A and antiprothrombin antibodies may be detected also by specific immunoassays. The capacity of several methodologies to detect antiphospholipid antibodies reflects chiefly their immunological and functional heterogeneity. Since most of the laboratory methods have not yet been standardized, the results of studies on the clinical relevance of antiphospholipid antibodies must be analyzed with caution. The association between antiphospholipid antibodies with peculiar clinical manifestations such as venous and arterial thrombosis, recurrent miscarriage, and thrombocytopenia, characterizes the so-called "antiphospholipid syndrome". Retrospective and cross sectional studies have confirmed the role of anticardiolipin antibodies and lupus anticoagulants as risk factors for both venous and arterial thrombosis, the most common clinical manifestations of the antiphospholipid syndrome. Prospective studies performed in different patient populations have confirmed the association between anticardiolipin antibodies and lupus anticoagulants with venous, and possibly, arterial thrombosis, although information on the predictive value of the various laboratory tests with respect to thrombosis is still limited. It is hoped that the development and standardization of assays that selectively identify antiphospholipid antibodies associated with increased risk of thrombosis will lead to therapeutic strategies able to prevent thromboembolic complications of the antiphospholipid syndrome. PMID- 9333317 TI - [Role of the liver in the regulation of glucose metabolism in diabetes and chronic liver disease]. AB - The liver plays a major role in the regulation of glucose metabolism: plasma glucose concentration is the result of peripheral glucose utilization and liver production. Several hormones, including insulin, glucagon, growth hormone, cortisol, and catecholamines contribute to the regulation of glucose metabolism by the liver. In this review, we examine hepatic glucose metabolism, in particular the actions of insulin and contrainsular hormones on glucose hepatic uptake and production in patients with diabetes or chronic liver disease. The most frequent patterns of hepatic involvement that take place during diabetes, i.e. nuclear glycogenesis, steatosis, portal fibrosis, and diabetic steatonecrosis, are discussed. Also considered are anomalies of glucose homeostasis observed in chronic liver disease, including glucose intolerance, diabetes, and hypoglycemias. There is a strong correlation between diabetes mellitus and the liver: diabetic patients have typical histological lesions, while several glucose metabolism alterations are commonly found in subjects with chronic liver disease. The pathogenesis of impaired glucose metabolism during chronic liver disease has not yet been fully understood: further clinical and experimental studies should clarify this issue. PMID- 9333318 TI - [Bilateral aneurysm of the common iliac vein: a case report]. AB - Vein aneurysm is a rare encountered lesion. We describe a case of extensive bilateral iliac vein aneurysm as an accidental finding during abdominal ultrasonography of a 39-year-old male marathon runner. Common iliac vein dimensions were respectively: right 39 x 42 x 60 mm and left 43 x 55 x 73 mm, with no signs of thrombosis or spontaneous echocontrast; mean flow velocity was 0.10 m/s with normal dynamics of venous flow. Furthermore we observed abnormal dilatation of the cava vein as well as the common femoral and popliteal veins. Other venous districts, detectable by color-Doppler ultrasound were normal. Known congenital or acquired conditions predisposing to aneurysm and/or ectasia were not detected. PMID- 9333315 TI - [Pancreatitis caused by Cryptosporidium parvum in patients with severe immunodeficiency related to HIV infection]. AB - Up to the present, pancreatic complications due to cryptosporidium parvum infection have been described only in a few patients with acquired immunodeficiency syndrome (AIDS). We report our experience with 3 subjects with AIDS who presented with acute or chronic pancreatitis related to cryptosporidiosis. All 3 patients had abdominal pain resistant to analgesics, increased serum amylase and abnormalities at both sonography and computed tomography. Endoscopic retrograde cholangiopancreotography revealed papillary stenosis in all patients; one patient also had stenosis of the Wirsung duct. Cryptosporidium was found in both bile and stool samples in all patients, while viral cultures were negative, even in the 2 patients who had cytomegalovirus retinitis. Endoscopic sphincterotomy temporally relieved abdominal pain in all patients, but did not prevent either acute or recurrent pancreatic inflammation. Several antibiotic therapeutic protocols were ineffective against the parasite. PMID- 9333319 TI - [Hypothyroidism with pseudo-ischemic and hypertensive clinical presentation: physiopathological and diagnostic considerations]. AB - Serious primary hypothyroidism, disclosed fortuitously through routine thyroid function test derangements, was found in a 40-year-old woman admitted to the hospital with a tentative diagnosis of ischemic heart disease. The clinical picture and electrocardiographic alterations of pseudo-ischemic heart disease associated with hypertension, particularly diastolic, may be the only significant manifestations of hypothyroidism. Substitutive hormone replacement therapy enables a good prognosis for children and young adults. A diagnosis of hypothyroidism should be considered during the initial evaluation of pseudo ischemic, hypertensive and hypercholesterolemic patients, even when no other signs or clinical symptoms of hormonal deficiency are evident. Particular attention should be paid to female patients, as they are much more frequently affected by thyroid pathologies. PMID- 9333320 TI - [Ethical aspects of randomized clinical trials]. AB - Randomized clinical trials represent the final, essential link between basic medical research and human health. However, their conduction presents very complex ethical problems, since the patient is the actual target of the experiment. Proper randomization, informed consent, and preliminary disclosure of results create deep ethical conflicts between the role of caretaker and that of impartial observer, both played by the same doctor. The dilemma reproduces the conflict between two different ethics. One is based on the inalienable individual rights stemming from the concept of man as an end in himself and not a means to an end. The other, derived from utilitarian philosophies, is based on the benefit for society as a whole. If we agree that randomized clinical trials represent the best method to test the validity of a new treatment, there is no easy solution. The dilemma could be solved by separating the role of the family doctor, committed to the best treatment possible for his patient, from the role of the scientist, committed to the progress of science and humanity. The former is involved in the treatment of individual patients, the latter in clinical and scientific experiments of a therapeutic nature. The patient may trade his rights to the best possible cure for the safety and the efficiency guaranteed by the scientific institution conducting the trial. Trials on relevant issues--expected to produce important results and impeccably designed scientifically--could be endowed with the ethics of science per se and this could be considered equivalent to the individual rights waived by the patient. PMID- 9333321 TI - Accumulating evidence suggests that several AB-toxins subvert the endoplasmic reticulum-associated protein degradation pathway to enter target cells. AB - Several AB-toxins appear to have independently evolved mechanisms by which they undergo retrograde transport from the cell membrane to the endoplasmic reticulum (ER). Recent insights into ER-associated protein degradation (ERAD) now provide clues as to why these toxins have selected the ER as the site of cell entry. We propose that they disguise themselves as misfolded proteins to enter the ERAD pathway. We further link the observation that these toxins have few, if any, lysine residues to the need to escape ubiquitin-mediated protein degradation, the ultimate destination of the ERAD pathway. The actual membrane translocation step remains unclear, but studies on viral immune evasion mechanisms indicate that retrotranslocation across the ER lipid bilayer may involve SEC61. Understanding the internalization process of these toxins opens new avenues for preventing their entry into cells. In addition, this knowledge can be exploited to create protein-based pharmaceuticals that act on cytosolic targets. PMID- 9333322 TI - Detection of an EPR multiline signal for the S0* state in photosystem II. AB - The S0* state was generated by incubation of dark-adapted (S1 state) photosystem II membranes either with the exogenous two electron reductant hydrazine and subsequent 273 K illumination in the presence of DCMU or by dark incubation with low amounts of the one electron reductant hydroxylamine. In agreement with earlier reports, the S1 and S-1 states were found to be electron paramagnetic resonance (EPR) silent. However, in the presence of 0.5-1.5% methanol, a weak EPR multiline signal centered around g = 2.0 was observed at 7 K for the S0* states generated by both procedures. This signal has a similar average line splitting to the well-characterized S2 state multiline EPR signal, but can be clearly distinguished from that and other modified S2 multiline signals by differences in line position and intensities. In addition, at 4 K it can be seen that the S0* multiline has a greater spectral breadth than the S2 multilines and is composed of up to 26 peaks. The S0* signal is not seen in the absence of methanol and is not affected by 1 mM EDTA in the buffer medium. We assign the S0* multiline signal to the manganese cluster of the oxygen evolving complex in a mixed valence state of the form MnIIMnIIIMnIIIMnIII,MnIIMnIIIMnIVMnIV, or MnIIIMnIIIMnIIIMnIV. Addition of methanol may be helpful in future to find an EPR signal originating form the natural S0 state. PMID- 9333323 TI - Coupled formation of an amidotransferase interdomain ammonia channel and a phosphoribosyltransferase active site. AB - Activation of gluatmine phosphoribosylpyrophosphate (RPPP) amidotransferase (GPATase) by binding of a PRPP substrate analog results in the formation of a 20 A channel connecting the active site for glutamine hydrolysis in one domain with the PRPP site in a second domain. This solvent-inaccessible channel permits transfer of the NH3 intermediate between the two active sites. Tunneling of NH3 may be a common mechanism for glutamine amidotransferase-catalyzed nitrogen transfer and for coordination of catalysis at two distinct active sites in complex enzymes. The 2.4 A crystal structure of the active conformer of GPATase also provides the first description of an intact active site for the phosphoribosyltransferase (PRTase) family of nucleotide synthesis and salvage enzymes. Chemical assistance to catalysis is provided primarily by the substrate and secondarily by the enzyme in the proposed structure-based mechanism. Different catalytic and inhibitory modes of divalent cation binding to the PRTase active site are revealed in the active conformer of the enzyme and in a feedback inhibited GMP complex. PMID- 9333326 TI - [The problem with assessing quality]. PMID- 9333325 TI - Endothelial nitric oxide synthase-dependent superoxide generation from adriamycin. AB - Adriamycin (or doxorubicin) is an active and broad spectrum chemotherapeutic agent. Unfortunately, its clinical use is severely restricted by a dose-limiting cardiotoxicity which has been linked to the formation of superoxide. Enzymatic one-electron reduction of adriamycin forms adriamycin semiquinone radical, which rapidly reacts with oxygen to form superoxide and adriamycin. In this way, adriamycin provides a kinetic mechanism for the one-electron reduction of oxygen by flavoenzymes such as NADPH-cytochrome P450 reductase and mitochondrial NADH dehydrogenase. We demonstrate here that the endothelial isoform of nitric oxide synthase (eNOS) reduces adriamycin to the semiquinone radical. As a consequence, superoxide formation is enhanced and nitric oxide production is decreased. Adriamycin binds to eNOS with a Km of approximately 5 microM, as calculated from both eNOS-dependent NADPH consumption and superoxide generation. Adriamycin stimulated superoxide formation is not affected by calcium/calmodulin and is abolished by the flavoenzyme inhibitor, diphenyleneiodonium. This strongly suggests that adriamycin undergoes reduction at the reductase domain of eNOS. A consequence of eNOS-mediated reductive activation of adriamycin is the disruption of the balance between nitric oxide and superoxide. This may lead eNOS to generate peroxynitrite and hydrogen peroxide, potent oxidants implicated in several vascular pathologies. PMID- 9333324 TI - Solution structure of the minor conformer of a DNA duplex containing a dG mismatch opposite a benzo[a]pyrene diol epoxide/dA adduct: glycosidic rotation from syn to anti at the modified deoxyadenosine. AB - Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants whose metabolism in mammals results in deleterious cell transformation. Covalent modification of DNA by diol epoxides metabolically formed from PAHs such a benzo[a]pyrene (BaP) provides a mechanism for the genotoxicity, mutagenicity, and carcinogenicity of PAHs. We had previously reported NMR evidence for a minor conformer of the duplex d(G1G2T3C4A5*C6G7A8G9).d(C10T11C12G13G14G15A16C17C18) containing a dG14 mismatch opposite a dA5* residue modified at the exocyclic amino group by trans addition to (+)-(7R,8S,9S,10R)-7,8-dihydroxy-9,10-epoxy 7,8,9,10-tetrahydrobenzo[a] pyrene [Yeh, H.J.C., Sayer, J.M., Liu, X., Altieri, A.S., Byrd, R.A., Lashman, M.K., Yagi, H., Schurer, E.J., Gorenstein, D.G., & Jerina, D.M. (1995) Biochemistry 34, 13570-13581]. In the present work, we describe the structure of this minor conformer (ca. 17% of the total conformer population). This represents the first structural determination of a minor conformer of a carcinogen-lesion DNA adduct. Two-dimensional NOESY, ROESY, TOCSY, and exchange-only spectra at 750 MHz allowed nearly complete sequential assignment of both conformers. In the minor conformer, the adducted base assumes an anti-glycosidic torsion angle whereas in the major conformer it assumes an unusual syn-glycosidic torsion angle. The aromatic hydrocarbon in the minor conformer is intercalated between dG13 and dG14, preserving the energetically favorable stacking interactions found in the major conformer. The major structural differences between the two conformers appear to be near the lesion site as evidenced by the large chemical shift differences between major and minor conformer protons near the lesion site; away from this site, the chemical shifts of the major and minor conformer protons are nearly identical. Because any of the conformations of benzo[a]pyrene diol epoxide-modified DNA may contribute to tumorigenic activity, structural determination of all conformations is essential for the elucidation of the mechanism of cell transformation initiated by covalent modification of DNA by PAHs. PMID- 9333327 TI - [Low dosage dopamine improves kidney function: current status of knowledge and evaluation of a controversial topic]. AB - The ability of low dose dopamine (1-3 micrograms x kg-1 x min-1) to cause selective renal vasodilation, to increase glomerular filtration rate, urine output, and natriuresis, is intuitively considered favourable. Dopamine, therefore, continues to be used in critically ill patients to preserve or improve renal function. Despite its application in a wide variety of disease states and in patients at risk of acute renal failure or with already decreased renal function, there is no conclusive evidence that "renal doses" of dopamine prevented acute renal failure or had any positive effect on patient outcome although it increased urine output and natriuresis consistently. Data from many clinical studies, however, are difficult to interpret due to small numbers of patients, the absence of control groups, the inability to exclude changes in cardiac output or to separate a diuretic effect of dopamine in the tubulus system from specific increases of glomerular filtration rate, and because of the variability of methods to determine renal performance (i.e. creatinine clearance, urine output, natriuresis, fractional excretion of sodium). Moreover, the routine use of dopamine is not innovous, since it may worsen gut ischaemia and suppress certain hormonal systems. Those who believe in the clinical benefits of "renal dose" dopamine argue that, even in the absence of an improvement in renal function, the maintenance of urine output could be useful in patients unresponsive to diuretics. Again, the clinical benefit of this diuretic action still needs to be shown. In conclusion, there is little justification for the routine administration of low-dose dopamine in patients at risk of renal failure. Large controlled clinical studies are urgently needed to determine whether dopamine improves renal function or prevents acute renal failure in patients at risk. PMID- 9333328 TI - [Effect of endoscopic surgery techniques in children on ventilation]. AB - The purpose of our study was to show the effects of laparoscopic procedures on the ventilation of children. We measured an increase of ventilation pressures (Pmax, Ppfat and Pmean) of 28, 35 and 48% respectively. petCO2 rose from 35 to 41 +/- 2 mmHg. There was no loss of body temperature in any child. Laparoscopic procedures as seen here for appendectomy or diagnostic exploration in children of 13.5 kg body weight or more caused no problems that were clinically evident. The increase of ventilation pressure could be attenuated by choosing the parameters of ventilation (e.g. flow, I:E ratio) in such a way that Pmax does not exceed 20 mbar. Alternatively, pressure-controlled ventilation may be used, adjusting petCO2 by ventilation frequency. In any case there must be a strict control of all possible side effects of laparoscopy, such as cutaneous emphysema or pneumothorax. PMID- 9333329 TI - [Effect of hydroxyethyl starch solution on kidney function in surgical intensive care patients]. AB - Hydroxyethyl starch is commonly used in resuscitation, anaesthesia and intensive care medicine. Increasing creatinine values in patients treated with hydroxyethyl starch have been described in some case reports. These have also been some clinical signs such as pain in the renal region, and swelling of kidney parenchyma, but no differential and sensitive parameters of renal function have supported these possible side effects of hydroxyethyl starch. Twenty-five patients were randomly allocated to a control and a treatment group. In the treatment group, patients received 12 ml per kg body-weight hydroxyethyl starch 10% (200/0.5) daily. A non-invasive diagnostic spectrum of renal function was performed. Specific tubular proteins (alpha 1-microglobulin, Tamm-Horsfall protein), glomerular parameters (immunoglobulin G, albumin), and the brush border enzyme acetyl-beta-glucosaminidase, were examined. Additionally, renal blood flow and glomerular filtration rate, Apache-II-score, breathing therapy, infusions, transfusion, and all medical interventions were documented. Both groups were comparable with regard to drug therapy. Apache-II-score, and fluid management. There were differences in tubular function between the two groups. Patients undergoing HES therapy showed increased excretion of alpha 1-microglobulin. Tamm Horsfall-protein and of brush border enzyme acetyl-beta-glucosaminidase. No significant differences were detectable in glomerular functions (glomerular filtration rate, renal blood flow), albumin, and IgG. Correlates of tubular damage after hydroxyethyl starch therapy were seen in intensive care patients. Many influences of intensive therapy act on renal function, and further studies with larger cohorts are necessary. With regard to the documented localisation of the tubular damage in the HES group, colloid therapy with hydroxyethyl starch in renal dysfunction should be monitored carefully by means of sensitive markers. PMID- 9333330 TI - [A new probe holding device for continuous bilateral measurements of blood flow velocity in basal brain vessels]. AB - The clinical relevance of transcranial Doppler sonography for the evaluation of cerebral perfusion and cerebrovascular regulatory mechanisms, as well as for the registration of embolic events, has increased considerably as a technique of non invasive monitoring. The continuous measurement of blood flow velocities in two different vessel segments, either ipsilateral or contralateral, is limited by intricate probe fixation and positioning for optimal insonation of the vessel on the one hand. On the other hand probes are displaced frequently during anaesthesiological measures, so that continuous registration during induction of anaesthesia is not always guaranteed. In view of these limitations, a new probe positioning and holding device has been developed and tested in a clinical study of patients undergoing cardiac surgery (n = 22). The newly designed probe positioning and holding device allowed the unilateral adjustment and continuous measurement of the blood flow velocity in the middle cerebral artery (Vmca) from anaesthesia induction to endotracheal intubation in all patients (n = 9). This was possible in only 61.5% (n = 8) of the patients who were monitored via conventional mode of probe fixation (n = 13). The new method rendered possible the positioning, insonation and measurement in two vessel segments in 77.8% of the patients, in contrast to 53.8% of the patients where the conventional technique was used. The newly designed probe holding device meets all standard requirements from the anaesthesiological viewpoint, and facilitates the perioperative application of transcranial Doppler sonography for non-invasive monitoring. PMID- 9333331 TI - [Quality assurance in intensive care medicine]. PMID- 9333332 TI - [External quality assurance in anesthesia]. PMID- 9333333 TI - [Patient data management systems--tools for quality management in intensive care medicine?]. PMID- 9333334 TI - [Modular development of a patient data management system for a surgical intensive care unit]. PMID- 9333335 TI - [Quality assurance in intensive care medicine. Results of a multicenter study in Germany]. PMID- 9333336 TI - [Interdisciplinary documentation, performance review and quality assurance in Austrian intensive care medicine]. PMID- 9333337 TI - [Status epilepticus late in pregnancy--eclampsia or subarachnoid hemorrhage?]. AB - After a largely inconspicuous pregnancy, a 31-year old primipara suffered from a status epilepticus in the third trimenon. The convulsions could not be terminated by emergency medical services, resulting in aspiration of gastric contents. Assuming eclampsia, an emergency caesarean section was performed immediately in a central hospital. Postoperatively, a pathological pattern of tendon reflexes was noticed. A CT scan revealed subarachnoid haemorrhage. The causal aneurysm of the right A. pericallosa was clipped subsequently. Eclampsia is the leading cause of epileptic seizures during pregnancy. However, a different aetiology should always be considered, especially if medical history does not reveal symptoms of pre eclampsia. PMID- 9333338 TI - [Noninvasive monitoring of cerebral oxygen saturation after severe craniocerebral trauma in a Jehovah's witness]. AB - Patients with severe head injury run a high risk of developing secondary cerebral defects. Various methods have so far been described which vary in their continuity, invasiveness and technical aspects for the early detection and treatment of complications. Within this group of patients Jehovah's witnesses pose a particular problem due to their restriction concerning therapeutical possibilities. That may mean an additional danger to the cerebral O2-delivery. This case report highlights on the value of continuous, non-invasive cerebral O2 saturation measurement and its combination with the fiberoptical monitoring of mixed venous O2-saturation. It has been possible to recognize episodes of imbalance between O2-delivery and its demand owing to a variety of causes. Other measured parameters were unable to reliably detect the development of critical complications. Further latrogenic loss of blood by laboratory tests was significantly reduced. Despite partially insufficient circulation and anemia with a hemoglobin value of 49 g/l we were able to discharge the patient from ICU without recognizable neurological sequelae. PMID- 9333340 TI - Regression estimator in ranked set sampling. AB - Ranked set sampling (RSS) utilizes inexpensive auxiliary information about the ranking of the units in a sample to provide a more precise estimator of the population mean of the variable of interest Y, which is either difficult or expensive to measure. However, the ranking may not be perfect in most situations. In this paper, we assume that the ranking is done on the basis of a concomitant variable X. Regression-type RSS estimators of the population mean of Y will be proposed by utilizing this concomitant variable X in both the ranking process of the units and the estimation process when the population mean of X is known. When X has unknown mean, double sampling will be used to obtain an estimate for the population mean of X. It is found that when X and Y jointly follow a bivariate normal distribution, our proposed RSS regression estimator is more efficient than RSS and simple random sampling (SRS) naive estimators unless the correlation between X and Y is low (/rho/ < 0.4). Moreover, it is always superior to the regression estimator under SRS for all rho. When normality does not hold, this approach could still perform reasonably well as long as the shape of the distribution of the concomitant variable X is only slightly departed from symmetry. For heavily skewed distributions, a remedial measure will be suggested. An example of estimating the mean plutonium concentration in surface soil on the Nevada Test Site, Nevada, U.S.A., will be considered. PMID- 9333339 TI - [Intensive care]. PMID- 9333341 TI - Use of two-stage test statistic in the two-period crossover trials. AB - For two-period crossover trials where the residual carryover can only exist in the presence of treatment effect, Willan (1988, Biometrics 44, 211-218) recommended use of the maximum test statistic that chooses the analysis with the larger test statistic corresponding to the parallel analysis and the crossover analysis. We construct two alternative two-stage test procedures, based on Grizzle's approach, that maintain the actual type I error rate at the desirable level. The power, accuracy, and precision of the analysis based on the modified two-stage procedures are compared to those based on the parallel analysis, crossover analysis, and the maximum test statistic. PMID- 9333342 TI - Finite mixture models for proportions. AB - Six data sets recording fetal control mortality in mouse litters are presented. The data are clearly overdispersed, and a standard approach would be to describe the data by means of a beta-binomial model or to use quasi-likelihood methods. For five of the examples, we show that beta-binomial model provides a reasonable description but that the fit can be significantly improved by using a mixture of a beta-binomial model with a binomial distribution. This mixture provides two alternative solutions, in one of which the binomial component indicates a high probability of death but is selected infrequently; this accounts for outlying litters with high mortality. The influence of the outliers on the beta-binomial fits is also demonstrated. The location and nature of the two main maxima to the likelihood are investigated through profile log-likelihoods. Comparisons are made with the performance of finite mixtures of binomial distributions. PMID- 9333343 TI - A note on multiple testing procedures in dose finding. AB - It is shown that, in some recently proposed multiple test procedures for dose finding (Tamhane, Hochberg, and Dunnett, 1996, Biometrics 52, 21-37), strong control of the familywise error is only guaranteed if order relations are assumed to hold among the means. Procedures that can be applied safely without assuming any restrictions on the means can be based on the classical many--one comparisons of doses with the zero dose. PMID- 9333344 TI - Optimum allocation of treatments for Welch's test in equivalence assessment. AB - As an extension of Welch's test to equivalence trials in a matched pair design, we determine the sample sizes n1 and n2 that maximize the power at given alternatives for a given total sample size n = n1 + n2. Although the optimal allocation is obtained asymptotically when the ratio of the standard deviations in both treatment groups equals the ratio of the sample sizes, numerical investigations show that this result does not hold for sample sizes where n < or = 25. For convenience, we provide tables containing the optimal combinations for the bioequivalence problem as well as for the problem of testing a clinically relevant difference. Results indicate that for small sample sizes, the optimum allocations of the treatments differ significantly in both testing problems, although they are asymptotically identical. In addition, we provide simple approximate equations that can be used for the determination of the required sample sizes to control a preassigned probability of type II error. PMID- 9333346 TI - A dynamic frailty model for multivariate survival data. AB - We consider the statistical modeling of data consisting of many study subjects with serially correlated multivariate survival responses. The (ordinary) frailty model handles the serial correlation in such data by introducing an unobserved multiplicative random effect term, called the frailty, in the hazard function. The frailties are often assumed to be identical for the survival times from the same unit. We have generalized the frailty model by allowing the frailties to vary stochastically with the indices. We have proposed a simple scheme to update the dynamic frailties. This approach assumes that the random effects are gamma distributed. At each occurrence, the two gamma parameters are updated according to the past information. In terms of their marginal distributions, the dynamic frailties form a multiplicative random walk. This approach results in a tractable likelihood. The small sample behavior of the MLE is studied via a simulation experiment. The model is then illustrated with a data set from an animal carcinogenesis experiment. PMID- 9333345 TI - Validity and efficiency of approximation methods for tied survival times in Cox regression. AB - Survival-time studies sometimes do not yield distinct failure times. Several methods have been proposed to handle the resulting ties. The goal of this paper is to compare these methods. Simulations were conducted, in which failure times were generated for a two-sample problem with an exponential hazard, a constant hazard ratio, and no censoring. Failure times were grouped to produce heavy, moderate, and light ties, corresponding to a mean of 10.0, 5.0, and 2.5 failures per interval. Cox proportional hazards models were fit using each of three approximations for handling ties with each interval size for sample sizes of n = 25, 50, 250, and 500 in each group. The Breslow (1974, Biometrics 30, 89-99) approximation tends to underestimate the true beta, while the Kalbfleisch Prentice (1973, Biometrika 60, 267-279) approximation tends to overestimate beta. As the ties become heavier, the bias of these approximations increases. The Efron (1977, Journal of the American Statistical Association 72, 557-565) approximation performs far better than the other two, particularly with moderate or heavy ties; even with n = 25 in each group, the bias is under 2%, and for sample sizes larger than 50 per group, it is less than 1%. Except for the heaviest ties in the smallest sample, confidence interval coverage for all three estimators fell in the range of 94-96%. However, the tail probabilities were asymmetric with the Breslow and Kalbfleisch-Prentice formulas; using the Efron approximation, they were closer to the nominal 2.5%. Although the Breslow approximation is the default in many standard software packages, the Efron method for handling ties is to be preferred, particularly when the sample size is small either from the outset or due to heavy censoring. PMID- 9333347 TI - Sequential analysis of censored survival data from three treatment groups. AB - In this paper, we propose a simple means of designing and analyzing a sequential procedure for comparing survival data from three treatments with the goal of eventually identifying the best treatment. Our procedure consists of the concatenation of two sequential tests, as is suggested by Siegmund (1993, Annals of Statistics 21, 464-483) for instantaneous normal responses. The first sequential test is a global test that attempts to detect an overall treatment effect. If one is found, the least promising treatment is eliminated and a second sequential test attempts to identify the better of the two remaining treatments. Although there are three different information time scales to consider corresponding to each pairwise comparison, we show that under certain conditions they may be approximated by a single time scale. This enables us to gain insight into the problem of censored survival data from the more easily understood case of instantaneous normal data. Also, it eliminates the need for intensive computations and simulations for the design and analysis of the procedure. PMID- 9333348 TI - Receiver operating characteristic studies and measurement errors. AB - A receiver operating characteristic (ROC) curve expresses the probability of a true positive (PTP) as a function of the probability of a false positive (PFP) for all possible values of the cutoff between cases and controls. Theta, the area under ROC curve, is a measure of the diagnostic ability of the separator variable. The usual nonparametric estimate of theta is shown to be based when the separator is measured with error. An expression for the largest-order term of the bias is found. The observed values and the measurement error variance are used to form a kernel estimate of the underlying distribution. These kernel estimates are used to estimate the bias. Monte Carlo simulation indicates that, for several families of distributions, the bias-corrected estimators have smaller bias and comparable MSE to the usual estimator. An application to the data of Clayton, Moncrieff, and Roberts (1967, British Medical Journal 3, 133-136) illustrates the technique. PMID- 9333349 TI - Analysis of multichannel patch clamp recordings by hidden Markov models. AB - Conventional methods of analysis do not allow the kinetics of patch clamped ion channels to be completely determined if more than one channel is present in the patch. This hinders investigations on small ion channels as well as on channel cooperativity and the homogeneity of channel populations. We present a method to extract the rate constants and current amplitudes for each individual channel from multichannel patches by a one-step procedure. For this purpose, the current record is modeled by the superposed Markov processes of the opening and closing of each channel that is contaminated by noise (Hidden Markov Model). Channel parameters are obtained by maximum likelihood methods. Because the parameters can be calculated directly from the unfiltered record, the dwell time and missed event problems are widely diminished. Confidence bounds for the estimated parameters are given. Statistical tests to decide whether channels switch identically and/or independently are introduced. The application of the method is demonstrated with simulated data. PMID- 9333350 TI - Small sample inference for fixed effects from restricted maximum likelihood. AB - Restricted maximum likelihood (REML) is now well established as a method for estimating the parameters of the general Gaussian linear model with a structured covariance matrix, in particular for mixed linear models. Conventionally, estimates of precision and inference for fixed effects are based on their asymptotic distribution, which is known to be inadequate for some small-sample problems. In this paper, we present a scaled Wald statistic, together with an F approximation to its sampling distribution, that is shown to perform well in a range of small sample settings. The statistic uses an adjusted estimator of the covariance matrix that has reduced small sample bias. This approach has the advantage that it reproduces both the statistics and F distributions in those settings where the latter is exact, namely for Hotelling T2 type statistics and for analysis of variance F-ratios. The performance of the modified statistics is assessed through simulation studies of four different REML analyses and the methods are illustrated using three examples. PMID- 9333351 TI - [Benign non-organ-related diseases of the retroperitoneal space]. AB - Because of the anatomic localisation of the retroperitoneal space, the detection and elucidation of pathology in the retroperitoneum calls for clinical acumen and the utilisation of imaging techniques. During the past two decades, efforts spearheaded by the work of M. A. Meyers led to an enhanced understanding of retroperitoneal anatomy and pathology. Conventional radiographic techniques are often incapable of detecting and/or characterising retroperitoneal abnormalities. Sonography may be limited by patient-dependent-factors. CT is unaffected by bowel gas and provides discrete cross-sectional images of the organs, fascial planes and retroperitoneal compartments, making it an ideal tool for assessment of retroperitoneal disease. In clinically stable patients MRT may be a useful modality for providing helpful and additional information in characterising retroperitoneal abnormalities. In this review article the diagnostic possibilities of benign not organ-related diseases of the retroperitoneum are described. This is intended to give the reader an insight into the etiology and distribution patterns of retroperitoneal fluid and gas collections as well as into diagnosis and differential diagnosis of benign retroperitoneal diseases. The diagnostic impact of the different imaging modalities is discussed. PMID- 9333352 TI - [Bladder and rectal infiltration by uterine carcinomas: the accuracy of CT and MRI compared to endoscopic procedures]. AB - PURPOSE: To establish the value of computed tomography (CT) and magnetic resonance imaging (MRI) in predicting bladder and rectum involvement in uterine carcinoma. MATERIAL AND METHODS: 6 different imaging signs (focal obliteration of perivesical or perirectal fat planes, area and angle of contact between uterus and bladder or rectum, asymmetric bladder or rectum wall thickening, evidence of intraluminal masses, and signal intensity of bladder or rectum wall on T2 weighted or contrast-enhanced MR images) were analysed retrospectively in 129 patients who underwent 92 CT and/or 64 MRI examinations. The data were correlated with intraoperative findings and the results of cystoscopy and rectoscopy. RESULTS: Asymmetric wall thickening, evidence of intraluminal masses and increased signal intensities of the bladder wall or rectum wall were valuable signs of infiltration (sensitivity 71-100%, specificity 91-96% and accuracy 89 97%). In 27 patients submitted to both imaging examinations MRI was somewhat superior compared to CT (p > 0.1) and yielded similar results as endoscopic procedures (accuracy of cystoscopy and rectoscopy of 90% and 94%, respectively). CONCLUSION: CT and MRI allow to predict involvement of bladder or rectum wall in carcinoma of the uterus with a similar accuracy as endoscopic procedures. PMID- 9333353 TI - [The hydro-MRT of chronic inflammatory bowel diseases]. AB - PURPOSE: Evaluation of hydro-MRI in the diagnosis of chronic inflammatory bowel disease (IBD). MATERIAL AND METHODS: 33 patients with suspected Crohn's disease or ulcerative colitis were studied prospectively. After distension of the colon by a rectal enema, breathhold MR imaging was performed during bowel relaxation. Results were compared to the clinical diagnosis, endoscopy, barium studies and histopathology. RESULTS: 24/24 patients with active Crohn's disease were correctly diagnosed by MRI. Conversely, MRI was positive in only 2/5 patients with ulcerative colitis. In 5 patients the presence of IBD was excluded. There were no false positives. CONCLUSION: Hydro-MRI is a very reliable modality in the diagnosis of Crohn's disease. In the differentiation of Crohn's disease from ulcerative colitis, hydro-MRI seems to be a promising imaging procedure. PMID- 9333354 TI - [Dynamic 31-phosphorus magnetic resonance spectroscopy of the m. quadriceps: therapy-induced changes in arterial occlusive disease]. AB - PURPOSE: The present investigation aimed at examining changes in muscle metabolism caused by treatment of arterial occlusive disease, using dynamic 31 phosphorus methods. METHOD: 32 patients with arterial occlusive disease were examined in a 1.5 T apparatus with a 6 cm surface coil before and after treatment. The metabolic changes in the quadriceps muscles were visualised during a 36 s phosphorus spectrum during rest, exercise (isometric and isotonic) and during a period of recovery. RESULTS: Vascular therapy resulted in a significant increase in the duration of both types of exercise during dynamic phosphorus spectroscopy (isometric exercise: 282 s against 199 s: p = 0.002, isotonic exercise: 575 s against 222 s; p = 5 x 10(-6). After treatment, exercise-induced changes in pH (7.00 against 6.94; p = 0.004 and 7.00 against 6.93; p = 0.02) and the ratio Pi/PCr (0.34 against 0.44; p = 0.002 and 0.36 against 0.50; p = 0.009) were significantly smaller than before therapy, using a similar amount of exercise. Recovery time of Pi/PCr (45 s against 82 s; P = 10(-5) and 42 s against 57 s; p = 0.01) and pH value (154 s against 181 s; p = 0.14 and 173 s against 214 s; p = 0.22) showed significant reduction after treatment. CONCLUSIONS: Dynamic 31-phosphorus magnetic resonance spectroscopy indicates increased mitochondrial oxidative capacity in the quadriceps muscles as evidence for increased oxygen supply to muscle tissue following vascular therapy. PMID- 9333355 TI - [A report on experience with a second-generation PACS installation in routine computed tomographic operations]. AB - PURPOSE: Our institute has been using a pilot SIENET-PACS system for the last five years; its advantages and disadvantages in daily use are described. MATERIAL AND METHODS: The SIENET system connects 2 CT, 3 MRT and 2 angiographic installations with 7 viewing consoles and with an optical archive. From January 1994 to 1997 approximately 19,000 patients were examined by CT and the results stored digitally in the PACS system. RESULTS: A representative analysis of selected patient data revealed problems in 12.8% of patients with regard to the radiological information system and also the hospital information system. While the digital data proved satisfactory, the production of digital images was excellent but time-consuming. The image archive was reliable but required greater capacity for our purposes. CONCLUSIONS: Manual retrieval of patient data presented problems and integration of the radiology information system and PADS is essential. Digital data gathering is an advantage. Digital imaging is too slow and should be improved by more suitable software. Integration of new archival storage methods and compatibility with the DICOM standard is, judging form our experience, essential. PMID- 9333356 TI - [The differentiation of adrenal gland tumors: an improvement in accuracy by a combination of fat-sensitive, T2-weighted and contrast-enhanced MR sequences]. AB - PURPOSE: To determine the diagnostic value of 3 MR sequences for the diagnosis of adrenal tumors. MATERIAL AND METHODS: Retrospective analysis of 32 adrenal tumors (10 malignant, 15 adenomas, 5 inflammatory, 2 phaeochromocytomas) examined by MRT (1.5 T) using T2 turbo spin echo, chemical shift imaging and dynamic contrast enhanced gradient echo sequences. RESULTS: Differentiation between malignant and benign adrenal tumours by means of T2-weighted sequences, chemical shift imaging and dynamic contrast-enhanced sequences achieved a sensitivity/specificity of 90%/73%, 100%/68% and 90%86%. Accuracy was 78%, 78% and 87%. By combining the three procedures, sensitivity/specificity/accuracy was improved to 100%/91%/94%. CONCLUSION: To differentiate adrenal lesions, the following procedures are recommended: demonstration of fat by chemical shift imaging is indicative of an adenoma. In the absence of any lipid content, T2-weighted and dynamic contrast enhanced sequences can demonstrate inflammatory lesions. PMID- 9333357 TI - [Mechanically detachable minicoils attached to a wire for superselective embolization]. AB - PURPOSE: Evaluation of clinical usability and effectivity of newly developed, mechanically applicable minicoils attached to a wire for super-selective embolisation. MATERIAL AND METHODS: The new embolisation coils have been used in 16 patients aged between 28 and 90 years for the following indications: 4 haemorrhages in cases of advanced carcinoma of the cervix, one false aneurysm, 7 traumatic lesions and 4 arteriovenous fistulas. The minicoils are made of platinum and are attached to a guide wire with a connecting hook. Application is via a microcatheter in coaxial technique. RESULTS: Percutaneous embolisation has been successful in patients. Additional surgery has not been required in any of them. Manipulation of the system is relatively easy and embolisation is made possible under controlled circumstances. In one case a minicoil disappeared into a peripheral vessel, but could be recovered percutaneously. CONCLUSION: The minicoils allow super-selective embolisation but, for reasons of cost, should be reserved to such vascular regions where the risk of misplacing must be kept at a minimum. Basic experience with embolisation techniques is indispensable for the application of this method. PMID- 9333358 TI - [Special technics for angioplasty of the brachiocephalic vessels]. AB - PURPOSE: Description of special techniques for retrograde angioplasty of cerebral arteries with protection of the brain. MATERIAL AND METHOD: During the last 3 years, 51 dilatations of cerebral vessels were carried out in 45 patients; in 8, special techniques were required. In cases with particular operative risks, angioplasty of cerebral vessels can be performed using various techniques (double catheter [8 cases], retrograde PTA [8 cases], kissing balloon technique [3 cases]). RESULTS: All stenoses were successfully dilated, using a local anaesthetic. One patient with a double stenosis of the common carotid artery developed severe headache and visual disturbances in the ipsilateral eye following dilatation of a tight stenosis with total remission of all symptoms within 24 hours. CONCLUSION: Stenoses of cerebral arteries where surgery would be difficult and associated with a high risk can be dilated by retrograde PTA using a double catheter technique with a low complication rate. PMID- 9333359 TI - [Digital storage phosphor mammography in a magnification technic: experimental studies for spatial resolution and for detection of microcalcifications]. AB - PURPOSE: Since the routine use of storage phosphor systems for mammography has been limited by its inadequate spatial resolution of 5 linepairs/mm, a combination of a magnification mammography technique with storage phosphor plates was investigated to detect microcalcifications. MATERIAL AND METHODS: A new mammography system with a microfocus tube using an anode of 0.05-0.12 mm allowed to obtain survey views of the breast with 1.7x magnification (m), and spot views with 4x magnification. The digital image receptor comprised a high resolution storage phosphor plate. To determine spatial resolution, contrast transfer curves were obtained, and the detection of microcalcifications was investigated by ROC (receiver operating characteristic) analysis. RESULTS: Spatial resolution for digital survey views (m = 1.7) was 8 linepairs/mm and for spot views (m = 4) was 18 linepairs/mm. ROC analysis demonstrated a significantly higher performance of the digital magnification technique compared to the conventional screen-film mammography technique. CONCLUSIONS: The limitations of digital mammography with respect to spatial resolution can be overcome by using a high magnification technique. PMID- 9333360 TI - [Intravascular MR imaging: the first in-vivo experiences in an animal arteriosclerosis model]. AB - PURPOSE: To evaluate intravascular MR imaging in normal New Zealand rabbits and hereditary hyperlipidaemic Watanabe rabbits (WHHL) with histological correlation. MATERIAL AND METHODS: The suprarenal abdominal aortas of two normal and two WHHL rabbits were examined by conventional angiography, high resolution MRT with a surface coil and intravascular MRT in a 1.5 T system. The intravascular reception coil consisted of a copper wire loop built into the balloon of an angioplasty catheter. The findings were correlated with histological examinations. RESULTS: Excellent spin echo images with a resolution of 78 x 156 microns were obtained in less than 4 minutes. The arteriosclerotic changes in the vessels of the WHHL rabbits could not be recognised angiographically. High resolution MRT with surface coils showed mural thickening but a detailed demonstration of arteriosclerotic lesions was possible only by means of high resolution intravascular imaging. There was good histological correlation. CONCLUSION: Arteriosclerotic lesions can be demonstrated in vivo by high resolution intravascular imaging. PMID- 9333361 TI - [MR tomographic temperature quantification at 1.5 T in vitro: a comparison of fast T1 maps and a phase-sensitive sequence]. AB - PURPOSE: To determine the value of fast T1-maps and a phase-sensitive sequence for temperature quantification with MR imaging. MATERIAL AND METHODS: The experimental setup allowed both homogeneous heating as well as laser-induced interstitial thermotherapy (Nd:YAG, 1064 nm) of tissue specimens (pig brain, liver and muscle) in vitro. A total of 60 experiments were performed. T1-maps were calculated by means of a multi-point-turbo-FLASH sequence. Additionally, the temperature dependent phase shift was determined with a 2D-FLASH sequence. RESULTS: The T1-relaxation times of four different aqueous solutions of gadolinium-DTPA (0.125-1.0 mmol/l) varied by less than 5%. During homogeneous heating, the T1-maps revealed a linear correlation (r > 0.98) between temperature and T1-relaxation times. The temperature coefficients were 11.0 +/- 0.42 ms/degree C. Variations of the linear correlation were observed during laser irradiation. There was only slight variation of the temperature coefficients of the chemical shift during homogeneous heating of different tissues (brain: 0.0098 +/- 0.0002 ppm/degree C; muscle: 0.0109 +/- 0.0003 ppm/degree C; liver: 0.0093 +/ 0.0002 ppm/degree C). The temperatures calculated during laser therapy based on the phase shift correlated strongly (r = 0.99) to the measured temperatures. CONCLUSION: Due to the considerable tissue independence and high accuracy, the phase mapping method is superior to T1-maps for monitoring thermal therapy modalities at 1.5 T in vitro. PMID- 9333362 TI - [CT-guided intervention by means of a laser marking and targeting aid]. AB - PURPOSE: The present study evaluates the use of a laser guidance system for CT guided intervention. MATERIAL AND METHODS: 94 cases of diagnostic biopsies and lumbar sympathectomies (54 cases with laser guidance system and 40 without) were compared. RESULTS: Using the laser guidance system, the number of control scans decreased by 30 to 50%, and necessary corrections of needle location were reduced by a maximum of 30%. The average target deviation of the needle decreased to less than 5 mm in 50% of cases. CONCLUSION: The laser guidance system is strongly recommended in CT-guided interventions for quality assurance and higher efficiency. The advantage is especially marked if the target area is small. PMID- 9333363 TI - [The computed tomographic demonstration of duodenal perforation caused by a fish bone]. PMID- 9333365 TI - [Shaken baby trauma: a case report on a problematic differential diagnosis in an infant]. PMID- 9333364 TI - [A solitary toxoplasmosis focus simulating a brain tumor as the first manifestation of AIDS]. PMID- 9333366 TI - [Odontogenic keratocysts--their imaging via MRT]. PMID- 9333367 TI - [MRT in tuberculous gonarthritis in childhood]. PMID- 9333368 TI - Genetic versus Environmental Determination of Human Behaviour and Health. Nobel symposium on the 150th anniversary of the Department of Paediatrics at the Karolinska Institute. 22-24 January 1996. PMID- 9333369 TI - [A model of integrated partial inpatient rehabilitation]. AB - This article deals with a new form of rehabilitation in Germany: the partly inpatient rehabilitation. This kind of rehabilitation comprises an offer for regular inpatient rehabilitation excluding overnight stay, breakfast and evening meal. In total the quasi inpatient rehabilitation is qualitatively equal to regular inpatient rehabilitation in respect of rehabilitative outcome. This new kind of rehabilitation contributes to a flexible and requirements-oriented health service in the German rehabilitation system. In this respect, "quasi inpatient rehabilitation" seems to be a valuable complement to traditional inpatient and out-patient rehabilitation, though quasi inpatient rehabilitation cannot generally replace traditional rehabilitation. PMID- 9333370 TI - ["Index for rehabilitation eligibility" for screening social insurance workers]. AB - The principle "priority of rehabilitation over early retirement" might be realised by a screening by which employees in need of rehabilitation are detected in time and rehabilitation measures are purposively started. With the "Index of Rehabilitation Need" we continued our efforts to develop an applicable screening tool on an epidemiological basis. To this end (1) longitudinal data were established by repeating an epidemiological survey of a population sample in the Nordenham/Brake region (T0 = 1975/76, T1 = 1992/93); (2) T0-variables were identified which correlated significantly with the events of early retirement and/or rehabilitation in the period of T0 to T1 (98 cases of early retirement/357 controls; 127 cases of rehabilitation/200 controls; 185 cases of early retirement of rehabilitation/270 controls) using bivariate and multivariate regression analysis; (3) significant T0-variables were used to construct a questionnaire index (based on self assessment of symptoms/complaints, consumption of medicaments, smoking, and work load--16 items), a medical examination index (based on clinical/laboratory findings and medical diagnoses--10 items), and an overall index (sum of both indices--26 items); (4) the index values were calculated for cases of early retirement of rehabilitation and controls of the cohort (185/270), for each index significant differences between cases and controls tested, and the screening characteristics of the overall index analysed; (5) possible reasons for incorrect classifications were examined using a subsample of cases and controls (n = 96/78), for which additional data on medical and work history, stressful life events, and attitudes towards rehabilitation had been collected. All indices showed significant differences between cases of early retirement or rehabilitation and controls. These differences proved to be stronger with the questionnaire and overall indices (p < .0000 each) than with the medical examination index (p < .0006). The overall index did not detect 18% of the cases in need of rehabilitation (false negatives). The proportion of the false positives was 14%; sensitivity and specificity amounted to 57% and 76%. The analysis of the subsample revealed only two possible and plausible reasons for incorrect classifications: the time span between the first survey and the year of early retirement as well as injuries. The index detected cases of early retirement or rehabilitation more easily where the time span between T0 and the year of early retirement was shorter. The index cannot detect cases of early retirement and rehabilitation caused by injuries between T0 and T1, since it is based on chronic disorders and stresses to be the reason for both events. With respect to the sensitivity and specificity of the index the relatively long prediction period needs to be taken into consideration--between T0 and the time of the events there could have been a period of up to 17 years. However, the objective of a screening is not to predict the long-term outcome but to preselect persons who are likely to need rehabilitation and should be invited to a socio medical examination in order to clarify their rehabilitation need and to start appropriate rehabilitation measures. The chance to detect true positive candidates and to exclude false negative candidates is essentially higher when the measurement of the predictors and the examination are carried out at the same time as has been shown in a former study. With regard to further proceedings we suggest to apply the index in a screening and to investigate the cost effectiveness and other aspects of the screening in a demonstration project. PMID- 9333371 TI - [Patient questionnaire for an epidemiologic cancer registry--results of an initial trial]. AB - For the first time a patient questionnaire was employed for an epidemiologic cancer registry to complete special history data routinely. After a successful pretest in 1995 it is being employed in the region Weser-Ems (Lower Saxony) since March 1996. The quality criterion completeness increased compared to the previously used sheet for history data, which had be filled in by the doctor. The quota of missing questionnaires is 10% and may be lowered by efficient measures. The occupational data should be used critically for comparison with other data sources (e.g. Bundesanstalt fur Arbeit [BfA]) or studies, because the code used by the BfA has been designed for other purposes and cannot be applied to investigations of occupational cancer risks. The geographical relation to cancer will be difficult to estimate because of the obvious mobility of the people. PMID- 9333372 TI - [Health status and medical care accessibility of single, homeless persons]. AB - The homeless population in Germany is continually increasing. Featuring prominently among those on the increase are women, young persons and homeless people from East Germany. Studies of the health of homeless individuals in recent years show that indices of illness are far higher for many disorders than for comparable groups who are housed. One result from a recent study by the University of Mainz (1994) was that more than 90% of homeless people urgently need medical treatment. According this research, the main health problems of the homeless are: cardiac disease (hypertension, CAD) (52.5%), skin disease (scabies, lice, leg ulcers, abscesses, pyodermias) and acute infections (50%), lower respiratory tract (COAD) (47.5%) and trauma victims (50%), followed by liver (30%), kidney (25%) and gastrointestinal diseases (GU) (20%). The problems of alcoholism and mental disorders of various sorts are added to this picture. Violence to homeless people is increasing. A lot of homeless people are multi morbid. The relationship between the time of homelessness and the state of illness was not linear. It was found that in the beginning of homelessness most of the homeless people were in a poor physical condition. The poor physical condition of homeless people does not stem from only one cause, but results from a combination of different factors: individual social conditions (social class; social relations; sedentary lifestyle), personal or family life crisis (life events and coping behaviour), the individual risk behaviour (for instance the bizarre sleeping accommodations, alcohol and cigarette consumption unemployment in a depressed economy, structure of the society (cutbacks in government welfare and social service programmes). As a result of bad experiences with existing medical institutions, homeless persons do not consult the doctor or too late. Many are afraid of large institutions; most are not members of a health insurance scheme (uninsured); and many are perceived in some sense to be "undesirable" as patients. Medical care offers for homeless people must be re-examined and changed appropriately in accordance with the requirements of the patients and the acceptability of the measures. Health care for the homeless is sorely needed. It is an urgent necessity to create special low-level acceptance medical care institutions. This health care service should be made available to homeless persons at the places where they gather (to set up a mobile medical service, medical streetwork, medical care ambulances). The interdisciplinary theme approach, which integrates the skills of physicians, nurses and social workers, is an invaluable strategy for establishing though and continuous care. Without good health, homeless people cannot resolve their other basic problems; and people simply cannot be healthy if they do not have a stable place to live. PMID- 9333373 TI - [Health promotion in Germany 10 years after the Ottawa Charter--exemplified by prevention of smoking]. AB - In Germany, there is no national preventive policy defining health problems, priorities and development of adequate intervention strategies including the factors of change. Thus, preventive potential remains unused. This is most evident for societal strategies, whereas individual strategies have been promoted when the responsibility for health promotion (h.p.) was resting with the sickness funds. In 1996, financing of h.p. through sickness funds was stopped. Redefining structures for preventive measures is overdue. Health authorities should play an active role in focussing on the coordination of health promotion. These arguments are discussed using the example of non-smoking campaign policies. PMID- 9333374 TI - [Polychlorinated biphenyls indoors: attempt at an assessment]. AB - The use of products containing PCB in buildings resulted in indoor air contaminations. The search for appropriate measures is hampered by several different uncertainties in the regulatory toxicological process. These refer to problems in exposure assessment including measurement approaches, the toxicological evaluation of health risks including the derivation of a tolerable intake and the use of TCDD toxicity equivalency factors and finally the legal consequences of health risk evaluation. Since no consensus on these aspects has been achieved, the regulatory consequences vary considerably from one German state government to the other. FAO/WHO have not been able to arrive at a satisfactory supranational recommendation of a tolerable intake for PCB. PMID- 9333375 TI - [Communication in environmental health protection and in environmental medicine counseling]. AB - Practically all municipal health authorities, including the Bremen health authorities, now offer advice with regard to healthy environmental protection, i. e. environmental-medical consulting activities. The majority of the consultantcy services deal with individual inquiries made by citizens. This is supplemented by counselling and discussions with groups in various contexts. It is frequent for extremely difficult counselling contents, situations and deficits in the conducting of the discussions by the consultants who are specialised in natural science subjects to coincide. Within the framework of a further training course designed for these specific needs, the staff of the Bremen health authorities, with specialist instruction, were successful in acquitting skill in the communication of risks, which is a qualification that should become self-evident also in this sphere. PMID- 9333376 TI - [Poisoning of a family by a mercury-containing ointment]. AB - Within the framework of a project entitled "Observation by Public Health Offices" we discovered a strikingly high mercury concentration of 30 micrograms in the urine of a 10-year old girl from an Albanian family formerly residing in the Kosovo area in Serbia. We investigated into the cause of this mercury load and found that it was also present in other members of the same family. Symptoms of mercury poisoning were evident to varying degrees in individual family members. Ultimately, we traced the source to a bleaching ointment containing mercury. On analysing the same we found a mercury content of 389 mg per g ointment. The use of this ointment with its extremely high mercury content had also contaminated the living-rooms. PMID- 9333378 TI - [Multiple chemical sensitivity (MCS)-syndrome]. PMID- 9333377 TI - [High percentage of isolated anti-HBc-positive persons among prisoners]. AB - PURPOSE: Is the result "isolated Anti-HBc" higher among prisoners than in the normal population and can testing for HBV-DNA clarify the results? PATIENTS AND METHODS: In Berlin, 519 prisoners were serologically tested in 1994 for hepatitis B and -C because of intravenous drug abuse and alcohol disease or signs of hepatitis. Beside virus antigen and antibodies, HBV-DNA was also measured by hybridisation technique or PCR. RESULTS: 50.3% of all individuals showed markers of hepatitis-B and 36.8% of hepatitis-C. 19.2% of persons with hepatitis B markers were positive for anti HBc only, i.e. more than twice as many than in the normal population. 90% of the isolated anti-HBc-positive Persons were also anti HCV positive, which is nearly double the number of individuals with other patterns of HBV markers. Half of them were tested for HBV-DNA. Whereas the hybridisation technique failed to detect HBV-DNA, 36% of sera were found positive by HBV-PCR. CONCLUSION: This study shows again that the result "anti-HBc alone" is relatively frequent especially among prisoners. This pattern often seems associated with concurrent HCV-infection and in one third of the cases correlated with a chronic hepatitis-B. The result of an isolated anti-HBc should therefore always lead to further testing of anti-HCV and HBV-DNA by PCR. PMID- 9333379 TI - ["Full shift light masonry work"]. PMID- 9333380 TI - [The public health office reports. Expert assessment of dyslexia--results of a survey]. PMID- 9333381 TI - [References on entero-hemorrhagic Escherichia coli]. PMID- 9333382 TI - [Analysis of psychiatric hospital cases in Hamburg 1988-1994--developmental trends, health care deficiencies and prospects]. AB - For the first time it was possible to study psychiatric inpatient treatment over a period of 7 years in a major German city (Hamburg) using data of 77% of all psychiatric inpatient cases collected by health insurance agencies. Among the most prominent results is the fact that 4 out of 8 illnesses with the highest sum of inpatient days of all treatment cases are psychiatric cases. These are schizophrenia, neurotic disorders, affective psychoses and alcohol abuse. Schizophrenia is the diagnosis which adds up to the highest amount of inpatient days in Hamburg hospitals. Of all psychiatric diagnoses, 35% show up in somatic departments, mainly internal medicine. This is especially true for alcohol and drug abuse, neurotic and personality disorders and organic psychoses. The greatest part of these cases were hospitalised for 0 to 3 days only, which points to the importance of crisis intervention provided by somatic departments. By introducing new offers of low threshold detoxification for drug abuse in psychiatry it was possible to increase the percentage of cases treated in psychiatry departments in the years 1993/94 as compared to 1988/89. During the same period the share of cases suffering from all kinds of psychoses decreased in psychiatry whereas the percentage of cases with drug abuse, neurotic and personality disorders rose. In nonpsychiatric departments, diseases seen in the context of alcoholism as well as neuroses and functional disorders prevail among the group of mental disorders. In internal medicine 6% of all cases are related with all kinds of addiction including its respective somatic consequences and 2 3% with neurotic and psychosomatic disorders. Looking at the amount of inpatient days 11.2% are spent for treating alcohol abuse, alcohol psychoses and diseases of liver and pancreas by patients of 15 to 65 years of age. In the light of these results it is suggested to set up psychiatric liaison-services in somatic departments, especially in internal medicine, to deal with psychosomatic and neurotic disorders and of course, alcoholism. This would help to lower hospitalisation costs. The introduction of motivational approach to the treatment of alcoholism in internal medicine departments appears warranted. Such changes of approach would result in new points of emphasis also in psychiatry. PMID- 9333383 TI - [Social health insurance for all: can we learn from Japan?]. AB - In the current German health care reform debate, the only foreign experiences discussed are from the USA or UK even though other countries have systems much more similar to that of Germany. Based on the German model of statutory health insurance. Japan has developed a similarly financed health system which has been quite successful in cost containment since the early 1980's. Unlike Germany, Japan has included the whole population in its statutory insurance scheme and refrains from private health insurance and upper limits on contributory income. Other features include the inclusion of pharmaceuticals in the Uniform Value Scale for all services, bi-annual revisions of that scale based upon utilization and technological innovation, government subsidies to health care which serve as a quasi global budget, and lastly a steadily growing GDP. Differences in the systems include the Japanese mixture of the ambulatory, in-patient and pharmaceutical sectors with physicians both in private practice and hospitals offering all three kinds of services. In-patient utilization is characterized by high numbers of hospital beds, very long lengths of stay, and few cases. Problems of the Japanese systems include the relatively low satisfaction of both population and patients, and the lack of quality assurance measures, clinical guidelines, and continuing education. PMID- 9333384 TI - [Need-based resource allocation--experiences with the RAWP formula in Great Britain]. AB - The RAWP formula used for resource allocation in Great Britain between 1976 and 1991 is a morbidity-oriented instrument of controlling, which has so far received only little attention in Germany. The development of this model was supported by the intention to intervene in the regional pattern of hospital supply by means of resource allocation and to refine it according to the guiding principles of equity and efficiency. The basic elements-regional population, average bed use, ICD chapter-specific SMRs-are discussed and the various modifications outlined. The RAWP formula's potentials of controlling resulted in a progressive reduction of the apparent disparities between regions in hospital supply, and knee was considered to be a "qualified success". The future development in the sense of an internal market addressed. PMID- 9333385 TI - [Stent implantation in coronary arteries--current status]. AB - Intracoronary stents have improved the safety and efficacy of transcatheter cardiovascular therapy. They can reverse acute vessel or graft closure during coronary intervention. Certain devices reduce the rate of restenosis and need for repeat interventional procedures compared with PTCA alone in selected patients. However caution is strongly recommended in generalising these findings to other situations than for which coronary stenting is considered. The rapid proliferation and application of this technique to very large groups of patients without supporting data should be reconsidered. Intermediate and longterm follow up date are not yet available. PMID- 9333386 TI - [Occupational medicine significance of hepatitis C in health care employees]. AB - Transmission of Hepatitis C virus (HCV) is similar to the one observed with hepatitis B virus. The most important route of infection in health care personnel is by needlestick injury. In the course of a literature review 44 publications on HCV prevalence or incidence among hospital employees were studied and an overall transmission incidence of 2.2% was found. As none of the publications dealt with the problem of chronic Hepatitis C, 245 persons with elevated transaminases occupied Freiburg University Hospital were tested for HCV antibodies. 3 out of 82 non-medical professionals and 19 out of 163 health care workers were anti HCV positive (RR = 3.22; p < 0.05). Charwomen in medical departments, nurses and dentists had a higher relative risk than physicians and technical assistants. As no HCV vaccine is available the only way of HCV-prevention is compliance with universal precautions. PMID- 9333387 TI - [Passive smoking in children up to 5 years of age]. AB - Passive smoking is a major health risk in young children. We investigated the percentage of children with mothers and/or fathers who reported regular smoking. Data are the national and regional health surveys of the German Cardiovascular Prevention Study (GCP) conducted from 1984 to 1992 in West Germany. 2538 mothers aged 25-40 years were included. The prevalence of passive smoking in children due to smoking mothers was 33.6% 55.4% of the children up to 5 years lived in households with at least one smoking parent member. In 23.4% of these households both parents were smokers. If only one member of the parents smoked this was in two out of three cases the father. 28.2% of mothers with a child younger than one year were current smokers. This prevalence rate increased with the age of the youngest child up to 35.6% for mothers, whose youngest child was 5 years old. Multiple logistic regression analysis was performed to investigate the association between smoking behaviour and the following variables: mother's age, social class, family status, community size and year of the survey. It was found that lower social class members, unmarried or divorced mothers and inhabitants of large cities reported significantly more often regular current smoking. These results underscore the importance of special intervention programs to reduce smoking in parents with young children. PMID- 9333388 TI - [Future developments in medicine: expert assessment]. AB - In recent years studies forecasting the future such as Delphi expert surveys were carried out in various Western countries to make predictions on future developments in medicine and in the health care system. The answers draw a homogeneous picture: Cancer and dementia research as well as the development of materials for implantation and artificial organs are-in context of the demographic changes-of enormous relevance, whereas basic research receives less attention. Due to various technological developments in telecommunication and in diagnostics (sensors), sociomedical care at home is supported and will become more frequent. PMID- 9333389 TI - [Catastrophy management in the large clinic. Bridges between fundamental research and rescue practice]. AB - In consequence of a fire that had broken out in the large general hospital centre of the University of Aachen, a working team was organised to systematise the experiences gained and to work out an internal scheme for handling catastrophic events. It was the declared aim of the working group to study and come to grips with innovative concepts of self-organisation in social systems and with cognitive trouble-shooting research to assess the relevance of actions based on these factors, the purpose being to complement and widen the range of conventional concepts of managing catastrophic events and the relevant security measures. This study discovered general agreement between the reflected experiences as seen from the viewpoint of the working team on the one hand, and the theoretical considerations of cognitive trouble-shooting research and the concepts of self-organisation in complex systems on the other. The innovative potential of knowledge for the internal management of catastrophic events derived in this manner from actions can hardly be over-estimated. PMID- 9333390 TI - [Studies of the development of refractive effects in intrastromal refractive corneal surgery with the picosecond laser]. AB - BACKGROUND: Picosecond laser intrastromal photorefractive keratectomy (ISPRK) aims at achieving a flattening of the central cornea by plasma-mediated tissue evaporation without affecting the anterior or posterior corneal layers. We investigated the laser-induced tissue effects to establish a functional relationship between laser parameters and tissue removal and to assess their influence on the healing process and long-term refractive changes. MATERIALS AND METHODS: A modified ISL 2001 System with a cone angle of 30 degrees was used for in vitro investigations of the laser effects in water and porcine cornea. Photographic methods were used to determine the plasma volume and the thickness of the laser-generated intrastromal bubble layer as a function of the pulse energy and the number and separation in which the pulses were applied (216 eyes). Histological evaluation was done by polarization microscopy (9 eyes). RESULTS: Polarization microscopy revealed only minor signs of thermal tissue damage. The maximum amount of tissue that can be evaporated without damaging the outer corneal layers corresponds to a layer about 10 microns thick. With a 6-mm optical zone, this tissue removal yields an immediate refractive effect of only 0.85 dpt. Stronger long-term refractive changes observed in animal experiments and clinical studies must thus be due to the healing response of the cornea. The healing response may be induced by mechanical distortion due to intrastromal bubble formation affecting about one third of the corneal thickness. CONCLUSION: Since the refractive effects are apparently strongly influenced by corneal healing, they are poorly predictable and can probably not be used for clinical purposes. PMID- 9333391 TI - [Effect of various ablation levels on endothelial density in the enucleated pig eye. Experimental study with the 193 nm Excimer laser]. AB - The effects of deep excimer laser ablations on the corneal endothelial cell count are still unknown. MATERIALS AND METHODS: In an in-vitro study the endothelial cell counts of 125 pig corneas were examined after excimer laser ablation. Five groups of 25 eyes reach, one control group and four groups with 3, 10, 20, and 35 D of ablation were examined. After the laser treatment, a corneoscleral disc was prepared and stored for up 5 days in an organ culture medium. The endothelial cell density of 5 corneas in each group was measured in a period of 5 days with a phase contrast microscope. RESULTS: Significant cell loss, 13-25%, was measured in each group after 5 days. There were no significant differences between the ablated corneas and the control group. CONCLUSION: Deep excimer laser ablations do not significantly reduce the endothelial cell count. PMID- 9333392 TI - [Optical coherence tomography of the cornea and the anterior eye segment]. AB - TARGET: The method of optical coherence tomography (OCT) was investigated regarding its suitability and limits for measuring the cornea and the anterior segment of the eye. Furthermore, the stromal expansion of thermally induced lesions in the cornea directly after irradiation was determined within the scope of the laser thermokeratoplasty (LTK). MATERIAL AND METHODS: With the experimental scanning OCT system, x-z sections of the anterior eye segment were made with an optical resolution of about 20 microns axially and 25 microns laterally. Freshly enucleated, tonicized porcine eyes were used as model eyes. Thermal lesions were applied with a continuously emitting laser diode (lambda = 1.86 microns) and various radiation parameters. Before and after coagulation, the cornea was viewed from limbus to limbus in a central OCT scan and the individual coagulation source was measured. RESULTS: Global and local changes of the thickness of the cornea as well as the distance between cornea and lens were measured with high precision. Thermal lesions in their expansion can be dearly presented and matching well with the histologically stained sections, but are not as exactly defined at the edges due to the limited optical resolution, as known from histological preparations. CONCLUSION: With the OCT method quantitative measuring of the anterior eye segment can be performed in vitro and with reduced resolutions also in vivo. Due to the qualitatively good correspondence regarding the dimensions of thermal damage of the cornea with histologically obtained morphometric results, this method can be used for supervision of coagulation directly after LTK as well as for examination of the individual healing process. PMID- 9333393 TI - [Imaging of laser thermokeratoplasty lesions by optical low coherence tomography and polarization microscopy after Sirius Red staining]. AB - BACKGROUND: Information on the extent and degree of the thermal effect produced is of great importance for control of the laser dosage in laser thermokeratoplasty (LTK) and for postoperative follow-up. We investigated on acute LTK effects which information images obtained by optical low coherence tomography (OCT) offer compared to those obtained by polarization microscopy. METHODS: Porcine eyes were irradiated through a 400 microns quartz fiber using light from a laser diode emitting up to 300 mW at a wavelength of 1.86 microns. Thermal lesions of varying strength were scanned using an experimental OCT device with about 25 microns lateral and 20 microns axial resolution. Histologic evaluation of the scanned areas was done by polarization microscopy after Sirlus Red staining, and similar lesions were also analyzed by TEM. RESULTS: Both methods differentiated three damage zones a transition zone, a zone of moderate coagulation, and a central zone of strong coagulation. In the transition zone, increased birefringence was seen in polarization microscopy, which correlated with increased light scattering seen in the OCT images. In the moderately coagulated zone, a decrease in birefringence was associated with an even stronger increase of the OCT signal. In the central zone, a loss of the fibrillar tissue structure was observed, which led to a complete loss of birefringence and a strong reduction of the OCT signal. CONCLUSIONS: Although OCT does not provide the detailed information on thermal changes of tissue seen by the histologic method, it offers information on the extent and degree of tissue changes without preparation artifacts and provides a non-invasive method of immediate and follow up control of LTK lesions. A quantitative analysis of changes in corneal thickness and curvature is much simpler than by a slit lamp. Time-resolved measurements of corneal light scattering may be used for on-line control of the laser-light dosage during LTK. PMID- 9333394 TI - [Temperature changes of the cornea by applying an eye bandage]. AB - BACKGROUND: The corneal temperature is not often measured, but it may be useful to evaluate the temperature-changing effect of the application of eye bandages. The reason for applying an eye bandage is to calm an inflamed eye and to provide mechanical protection. Everyone knows that some patients have more complaints after an eye bandage has been applied. This phenomenon might be caused by the inflamed eye being warmed up by bandage application. MATERIALS AND METHODS: In 40 apparently healthy subjects 24 +/- 1.93 years of age we examined the temperature changes caused by two different types of eye bandage. The measurements were done with a Jeol infrared camera. Twenty probands received a monocular bandage consisting of a perforated plastic cap and another 20 subjects a monocular mull bandage with a perforated plastic cap. RESULTS: The mean corneal apex temperature was 32.05 +/- 0.74 degrees C. Both bandages caused the corneal temperature to go up considerably. The perforated plastic cap increased the mean apex temperature by 0.58 +/- 0.48 degree K and the combined mull bandage by 1.15 +/- 0.57 degrees K (P < 0.05). CONCLUSIONS: Application of an eye bandage increases the corneal temperature significantly. This is known to change enzyme activities and to cause prostaglandin liberation and pain. A change in the bacterial spectrum may result. Cooling as a universal principle in antiphlogistic therapy might be a supplementary therapy in treating sterile, but inflamed eyes. PMID- 9333395 TI - [Preventive systemic cyclosporin A after keratoplasty at increased risk for immune reactions as the only elevated risk factor]. AB - BACKGROUND: In this retrospective study our aim was to evaluate the effectiveness of systemic cyclosporin A (CsA) after keratoplasties with an elevated risk for immune reactions as the only elevated risk factor. PATIENTS AND METHODS: Between November 1986 and June 1994, 1121 penetrating keratoplasties, 646 normal-risk and 475 high-risk keratoplasties were performed. In 130 out of the 475 high-risk keratoplasties an elevated risk for immune reactions was the only elevated risk factor. Twenty-six of these 130 high-risk keratoplasties were treated with systemic CsA. RESULTS: In the high-risk group keratoplasties with an elevated risk for immune reactions as the only elevated risk factor no permanent graft failure occurred with CsA (100% clear grafts). Without CsA the percentage of clear grafts in this high risk group was only 71.7% according to Kaplan Meier 3 years postoperatively in contrast to 86.0% in normal-risk keratoplasties. The differences between these three groups were statistically significant. In the high-risk group keratoplasties with on elevated risk for immune reactions as the only elevated risk factor more immune reactions occurred than without CsA or than in normal-risk keratoplasties. However, these immune reactions were mostly of the benign chronic types. CONCLUSIONS: Systemic CsA considerably improves graft prognosis after high-risk keratoplasties with an elevated risk for immune reactions as the only elevated risk factor. With CsA application we observed a significant shift from acute to chronic immune reactions, which respond much better to topical steroids. PMID- 9333396 TI - [Keratoprosthesis for experimental surgical use of donor eyes after corneal removal]. AB - BACKGROUND: Keratoprosthetic devices have been introduced for short-term use during surgical interventions and for long-term therapeutical use. The keratoprosthesis presented here was developed for the experimental use of human donor eyes after removal of the cornea. MATERIALS AND METHODS: In a step-by-step modification process, the main constructive elements of the keratoprosthesis were modified and tested for handling, operative stability and optical quality. PMMA was used as basic material. RESULTS: The final model was a two-piece device consisting of a screw (8.0 mm long, 14.5 mm in diameter) with an ample base (17.5 mm in diameter) for the introduction into the anterior segment following a corneal removal 15.0 mm in diameter. A threaded outer ring fixes the scleral rim against the screw. Lateral boreholes enable the devices to be handled by ordinary surgical forceps. CONCLUSIONS: The keratoprosthesis proved to be a useful and efficient aid in experimental and nearly realistic vitreoretinal surgery with human donor eyes after removal of the cornea. PMID- 9333397 TI - [Long-term prospective study of the development of corneal astigmatism in no stitch cataract surgery]. AB - BACKGROUND: Several studies have confirmed, that the no-stitch technique results in early stabilization of astigmatism. In these papers, however, the follow-up was quite short. Former studies about sutured corneoscleral incisions have shown that even after some years astigmatism increased. Therefore, in this prospective study we investigated the change of induced astigmatism 5 years postoperatively. METHODS: The prospective study included 66 patients with scleral tunnel incisions who were monitored up to 5 years. Mean age was 74.3 +/- 10.7 years. The incision length of the scleral tunnel was 7 mm. The postoperative astigmatism was measured with keratometry after 1 day, 3 weeks, 1 year and 5 years postoperatively. RESULTS: The average postoperative induced astigmatism (PIA) as measured with a keratometer was after 1 year 0.95 +/- 0.83 D. After 5 years PIA was 0.96 +/- 0.48 D, and therefore no significant statistical difference could be shown. At the first postoperative day (0.94 +/- 0.73 D) the induced astigmatism was virtually stable. There was also no statistically significant difference in absolute astigmatism. After 1 day the absolute astigmatism was 0.91 +/- 0.73 D and after 5 years 0.96 +/- 0.61 D. CONCLUSIONS: The contrast to former sutured corneaoscleral incisions, the no-stitch technique in cataract surgery yields stable postoperative corneal astigmatism. PMID- 9333398 TI - [Development of a standardized evaluation system for cataracta complicata in retinitis pigmentosa]. AB - INTRODUCTION: Retinitis pigmentosa (RP) is associated with the formation of a posterior subcapsular cataract (PSC). As only a small part of the crystalline lens is usually affected, it is sometimes difficult to determine to what extent the visual loss is caused by the PSC alone. PATIENTS AND METHODS: The methodology was developed in analogy to a scoring system for posterior capsule opacification by Tetz et al. Following dilation of the pupil, standardized photographs of the anterior segments were obtained utilizing a Zeiss photoslitlamp model 40 SL/P. The PSC was scored by evaluating retroillumination photographs. The individual PSC index was calculated by multiplying the density of the opacification (graded from 0 to 4) by the area involved in the central 4 mm zone of the pupil (calculated between 0 and 1). For testing the reliability of the evaluation system in part 1 of this study, 11 RP patients with different grades of PSC were examined by three independent observers. In part 2 of this study 37 eyes of 24 RP patients, aged 47.2 +/- 11.8 years, were evaluated and the PSC index was correlated with different parameters (visual acuity, age, visual fields, eletroretinography). RESULTS PART 1: The cataract-density grades were between 1 and 4 in the 11 patients. In relation to the central 4-mm pupillary zone between 13 and 100% of the area were opacified. Cataract indices (density x area) were between 0.13 and 4.0 (Mean values: Examiner 1:1.41 +/- 1.49; Examiner 2:1.28 +/- 1.46; Examiner 3:1.22 +/- 1.44; differences not significant: P = 0.77). PART 2: After an average duration of RP of 23 years, the average cataract index of the 24 patients was 1.72 +/- 1.35. There was no correlation between cataract index and ERG or visual fields (r < 0.2; P > 0.4); however, there was a good correlation to visual acuity (r = -0.72; P = 0.0001). Patients with early onset of RP (before 20th year of life) presented on average with an higher cataract index (2.06 +/- 1.67) compared to patients with late manifestation (0.61 +/- 0.44), but equivalent duration of RP. CONCLUSIONS: The evaluation system offers a reliable and reproducible method for measuring PSC density and extension in RP patients. The method can serve as a useful tool for documenting PSC development and help to define the indications for cataract surgery in RP. PMID- 9333399 TI - [Changes in corneal thickness and endothelial cell density after cataract extraction using phacoemulsification]. AB - Corneal metabolism is reduced after cataract extraction. PATIENTS AND METHODS: In a prospective study the corneal thickness and endothelial cell count of 48 patients were examined after phacoemulsification. Corneal thickness was measured with an ultrasound pachymeter and the endothelial cell count with a contact endothelial camera at the 12 o'clock position and in the corneal center before and 4 weeks, 4 months and 1 year after operation. RESULTS: One year after the operation, corneal thickness increased about 9% in the 12 o'clock position and about 12% in the corneal center. The endothelial cell count decreased about 27% in the 12 o'clock position and about 18% in the corneal center. We measured a significant correlation between cell loss and age at both points. Concerning the corneal thickness, no significant correlation was found. CONCLUSION: After cataract extraction corneal metabolism is reduced. The endothelial cell count or corneal thickness can be used as an indicator of the corneal trauma resulting from the operation. PMID- 9333400 TI - [Contrast vision and contrast sensitivity after binocular implantation of multifocal or monofocal intraocular lenses]. AB - BACKGROUND: Multifocal intraocular lenses show a reduction of contrast due to the simultaneous projection of different images to the retina. In this study we examined if there is also a loss of contrast in bilateral implantation of multifocal lenses. MATERIAL AND METHODS: We examined 22 patients with bilateral AMO ARRAY multifocal intraocular lens and compared these to 20 patients with bilateral monofocal intraocular lens. We performed a monocular and binocular examination of contrast acuify by means of Regan's contrast charts and contrast sensitivity by means of B-VAT-II-SG-Video-acuity-tester. RESULTS: Monocular examination of contrast acuity showed significant superiority of the monofocal intraocular lens at the lowest contrast. Bilateral examination of contrast acuity did not show any significant difference. Monocular contrast sensitivity of the monofocal intraocular lens was significantly superior to the multifocal intraocular lens at two spatial frequencies, but under the bilateral condition there was only a significant difference between the two lenses at the highest spatial frequency. CONCLUSIONS: Bilateral implantation of multifocal intraocular lenses enables contrast acuity and contrast sensitivity that comes very close to the performance of the monofocal intraocular lenses. PMID- 9333401 TI - [Scientific role of German ophthalmology in the European telecommunication project OPHTEL]. AB - BACKGROUND: In Denmark, France, Germany, Great Britain and Italy, the OPHTEL project combines clinical centers of ophthalmology and internal medicine, an institute for medical informatics and health services research, a publishing company and different industrial partners in the EDP market. AIMS: With the aid of visual telecommunication and rapid data transfer, methods and conditions will be developed and proved so that any physician can very easily obtain sufficient information for treating his patient. Thus, the regional differences in the quality of structured health service (e.g., urban/ rural) will be overcome throughout Europe. SCIENTIFIC TASKS: A multilingual diagnostic and therapeutic thesaurus has to be worked out in order to create standards for communication and quality control. Based on literature, images and image analysis in a knowledge based data bank, a monitoring system (containing watch-dog functions) and the basic aspects of an ophthalmological patient/disease register will be investigated. (In parallel, a technical development of synchronous and asynchronous telecommunication between eye physicians is taking place in close cooperation with the regional Bavarian project Teleopathalmology in Bavaria on line). RESULTS: State of the art 6 months after starting the project:the knowledge-based image data bank has been founded and also an ophthalmological 8 language thesaurus and definition standard. All data transfer lines are installed. DISCUSSION: The project is taking place amid diverging sections of medicine: ophthalmology and internal medicine, health politics and data protection, individual treatment and common interest (health care), product management and office organization. Thus, the scientific quality of the transferred ophthalmological content must undergo sophisticated controls. FUTURE STEPS: Intense cooperation with the big German associations for ophthalmology (DOG, BVA) and the European ophthalmological societies concerning EDP, classification and quality control. PMID- 9333402 TI - [Mitomycin as local therapeutic drug in ocular neoplasms]. PMID- 9333403 TI - [Progressive, unilateral vision loss with changing nonspecific visual field findings. Chromophobic anterior pituitary adenoma]. PMID- 9333405 TI - Bibliography. Current world literature. Sensory systems. PMID- 9333404 TI - [Albinism. Current clinical and molecular genetic aspects of an important differential congenital nystagmus diagnosis]. PMID- 9333406 TI - [A new family intervention method]. AB - PROBLEM BEING ADDRESSED: Family medicine residents have difficulty developing the complex skills needed to work with families. OBJECTIVE OF PROGRAM: To develop an educational program to teach family medicine residents at Laval University a new type of family intervention technique, based on the systems approach, using a practical and interactive method. MAIN COMPONENTS OF PROGRAM: Using Brien's systematic planning model, the authors developed and tested a series of six interactive workshops on three themes. The first theme aims to motivate residents by showing them why it is important to use a systems approach with their patients. The second theme explores a family situation in an individual interview. The third enables residents to conduct a family interview. The whole program has been tested and evaluated. All of the materials needed to teach these skills are contained in a trainer's guide and a video cassette. CONCLUSION: This educational program's originality lies in its combination of the systems approach and interactive training for residents. The program could easily be presented as a continuing medical education activity. PMID- 9333407 TI - [Water as a stabilizer and plasticizer of the globular structure of lysozyme]. PMID- 9333408 TI - [Benzimidazolium derivatives with delocalized charge in the cation group as reversible inhibitors of cholinesterases of different origin]. PMID- 9333409 TI - [Biochemical characteristics of food and uterine myoma]. PMID- 9333410 TI - [Expression of calcium-binding proteins parvalbumin and calbindin in neurons of the neocortex grafts]. PMID- 9333412 TI - [A role of early olfactory experience in the individual recognition in gray rats (Rattus norvegicus)]. PMID- 9333411 TI - [A study of penetrance of Drosophila melanogaster forked gene]. PMID- 9333413 TI - [Study of transcription of the transposable element burdock at the different stages of Drosophila development]. PMID- 9333414 TI - [Study of the biological activity of the water-soluble fullerene complexes]. PMID- 9333415 TI - [Hypothermia induced by high pressure of nitrogen]. PMID- 9333416 TI - [Insight phenomenon in the regulation of time of performance of behavior acts in monkey (Macaca mulatta)]. PMID- 9333417 TI - [Possible involvement of histamine in humoral regulation of the spontaneous contractile activity of the chick embryo amnion]. PMID- 9333418 TI - [Comparative validation of manual and automated methods for mixing and volume control of total blood samples]. AB - During blood collection, agitation and volume limitations are critical to ensure thorough mixing of the blood with the anticoagulant and obtention of the predetermined volume. These 2 factors are essential to prevent blood activation and to obtain well standardized blood products. The objective of this study was to compare the quality of the blood collected using 2 types of collection method: tripping of a scale at a predetermined volume limit of 450 mL in the presence of manual agitation, and the 3 blood collection monitors currently available in France. A minimum of 100 collection procedures was performed for each of the 4 methods tested. Results were found to be equivalent using either the manual or the automated procedures with regard to both the accuracy and reproducibility of the blood volumes obtained and the collection times and flow rates. The characteristics of the red blood cell concentrates, platelet concentrates and plasma units prepared from the first 30 collections of each group were assessed and compared to regulatory requirements. The quality of all these products was found to be comparable to that currently observed at quality control and no product was rejected at the release control for reasons of poor collection. An assessment of the practicability of the different methods showed that the automated devices are subject to practical difficulties involving transport and battery loading. In addition, the cost of this equipment is approximately 5 times higher than that of the scales. In conclusion, the results of this study show that in our hands, no significant advantage could be expected from the use of automated blood collection monitors as compared to simple scales with manual mixing. These results further raise the question of the applicability to labile blood products of the comparative validations currently accepted in the pharmaceutical industry, in order to allow the use of correctly validated alternative methods. PMID- 9333420 TI - [Screening and early detection in blood transfusion: when are they indicated?]. AB - Screening is proposed to allow an early intervention concerning a diseased individual, but its public health consequences are seldom considered. We propose criteria to judge whether a screening program can be associated to a benefit to patients or society. These criteria refer to the magnitude of disease, the characteristics of the pre-clinical stage, the availability of reliable and valid tests, the effectiveness and risk for all individuals, whether they are diseased or not, and acceptability to the health-care system and to individuals. We illustrate the application of these criteria to screening of human immunodeficiency virus among blood donors, hepatitis among recipients of labile blood products, and bacterial contaminations among febrile recipients. These criteria should be considered in decision analyses including alternatives to screening and all aspects of safety regarding patients and population. PMID- 9333419 TI - [Screening for markers of blood-borne diseases in donated units collected in France from 1993 to 1995]. AB - A decrease of positive donation rates for antibodies to HIV, to HCV and for HBs Ag has been observed between 1993 and 1995 both in first-time and regular donations. In first-time donors, the most important decrease has been observed for HIV and in regular donors for HCV and HBs Ag. The interval between the negative and the positive donations was inferior to 1 year for 50% of regular donors and was superior to 2 years for 20 to 30%. About 30% of HIV positive donations were positive for other markers: 23% for anti-HBc and 10% for anti-HCV. About 40% of HCV positive donations had an elevated ALT level. Risk factors related to HIV heterosexual transmission appear to be the most difficult to identify during the donor selection and the least often associated with other markers. In recently HCV-infected donors, the main risk factors were IV drug addiction (25%) and nosocomial infection (30%). The major HTLV risk factor was directly or indirectly linked to the Caribbean area. Important differences between continental France and overseas territories were observed for HIV and HBs Ag rates. PMID- 9333421 TI - [Neuroimmunology of Alzheimer's disease]. AB - Alzheimer's disease (AD) is the most frequent cause of premature, irreversible cognitive decline in man, the consequence of which is complete psycho-social incompetence. 60% of all dementias are of the AD type, with its prevalence increasing logarithmically with age. Statistics from countries where average life expectancy is around 70 years show that 1% of the total population suffers from AD. If current demographic trends do not change, 40% of the population will be over 60 in 20 to 25 years and the number of AD sufferers will increase 5 times. AD is the fourth commonest cause of death in humans, after heart attack, cancer and stroke. We do not have any cure or efficient preventive measures against AD. The duration of the disease varies from 4-12 years and it is always fatal. Pathological characteristics of AD are neurofibrillatory tangles and senile plaques. Despite the identification and molecular characterization of the pathological forms of tau and beta-amyloid found in these pathological features, little is known of the etiology and the pathogenesis for AD. The main goal of current research is the identification of interactions between the nervous and immune systems which may be involved in the neuropathology and pathogenesis of AD. Namely, the identification of those sequences of molecular, cellular and systemic mechanisms which lead to the fatal neuronal degeneration which ultimately results in dementia. Understanding AD pathogenesis may lead to early AD diagnostic assay and new potentials for therapy. PMID- 9333422 TI - [Control of autoimmune processes by natural and other non-harmful methods]. AB - At present an increase of some autoaggressive diseases can be observed. The commonly used treatment consists of the administration of some immunosuppressive drug of some hormonal preparations. This type of therapy is accompanied by some undesired side effects, as these drugs influence also some other cell systems besides the immunologically active cells. These drugs are also known to lower the resistance to some intercurrent infections. Due to these undesired side effects some naturally occurring factors are introduced into the therapy. These are e.g., TGF-beta, or some interleukins (IL-10 etc.). In our department and immunosuppressively acting substance was isolated from DHL which had the ability to inhibit the AA (adjuvant arthritis) in rats. In humans this SF (suppressive factor) stimulates the CD 8+ cells which are known to have suppressoric activity. This SF was successfully applied in some autoaggressive diseases, e.g., atopic eczema, multiple sclerosis, some polyradiculoenuritis, amyotropic lateral sclerosis etc. In this paper the results in the ALS patients are given. Amongst other possibilities of the therapy the application of antilymphocyte sera, monoclonal antibodies to some CD markers of lymphocytes and some methods of hyposensitizations of tolerance induction are mentioned. Further, an original method using antigen bound to isosoluble carrier is described. This administration of encephalitogen bound onto Sforon (polyacrylate spheres) sis not only inhibit the EAE manifestations but also enable the survival of guinea-pigs which had already manifested the clinical signs of EAE. PMID- 9333423 TI - [Cerebral infarct and the immune response]. AB - There are only few data available regarding the immunological mechanisms for cerebral infarction. The aim of this study was to find out the humoral and cell mediated immunity under the conditions of focal brain ischemia (CI). As a method for humoral immunity, the complement consumption test against a panel of 8 antigens, quantitative analysis of immunoglobins and fractionized sedimentation of erythrocytes were used in the group of pts with CI, and the group of atherosclerotics (AS) and hypertonics (VH), potential victims of focal brain ischemia. It was found that the occurrence of antibodies against the whole panel of antigens in the group of CI is significantly higher as compared with the healthy controls, but it is lower than that in the group of AS and VH. The occurrence of antibodies exclusively against only brain antigens and that in CSF is similar. No correlation to the location of ischemic lesion and the degree of neurological deficit score was found. These findings didn't change in 2 and 4 weeks as well as in 1 year after the onset of CI. The quantitative analysis of immunoglobins revealed statistically higher levels of IgA and lower levels of IgM in comparison with the controls. IgG were higher, but without statistical significance. Statistically significant higher levels of all immunoglobins in CSF were found. As similar trend of changes found also in the group of AS and VH. These results of humoral immunity confirmed by the results of fractionized sedimentation of erythrocytes with EP. The results can be interpreted as a possible change or disorder of central regulation of immunizing processes due to the latent (in AS and VH) of manifest (in CI) lesions of the brain. But the quality and quantity of this response might have been affected by the entire case history of the patients who survived cerebral infarction. The changes in immunity response of the organism in CI was shown also in cell-mediated immunity. The results a statistically significant increase in stimulatory (SI) as well as in immunoregulatory (IRI) indices in stroke patients under the age of 40. These findings didn't change 2 and 4 weeks after the onset of CL. An increase in IRI was due to the increase in Th lymphocytes. In the immune response of the organism in CI, the antiphospholipid antibodies (aPLs = anticardiolipin antibodies (aCL) and lupus anticoagulant--LA) play an important role. aCLs were present in 9.8% of the first stroke pts when compared to 4.3% in controls. The most common isotype of the antibodies we IgG. Of all first-stroke pts who were aCL positive only 8% had no other stroke risk factors (atrial fibrillation, diabetes, hypertension and other). aCLs are an important risk factor for the first stroke, mainly in the young, but also in the elderly. The presence of aCLs increases the risk for recurrent strokes. aPLs are not necessarily associated with the specific location of clinical stroke syndrome but they are in significant correlation to the occurrence of multiple strokes on CT (30:18%). None of the initially aCL-negative patients became aCL-positive during the time course of CI. These data support the idea that aCLs play a causal role in stroke (PROPTER HOC changes) rather than vice versa (POST HOC changes). From the therapeutic point of view, currently there do not exist any good treatment guidelines for preventing the second stroke. The analysis of HLA. antigen showed an increase in some HLA (A2, A28 etc.) and a decrease in others (A3, A9 etc.) in comparison with the controls. This might refer to the participation of genetic factors in the onset of CI. PMID- 9333424 TI - [Encephalopathy in AIDS--increased formation of beta-chemokines in monocytes after HIV-1 virus infection: mechanisms of CNS involvement]. AB - The characteristic trait of the family of lentiviruses (Retroviridae) which includes the human immune deficiency virus (HIV), is the tendency to cause a subacute neurologic disease in their animal host. The neuraxis can be inflicted at all its levels. In the advanced stage of HIV disease, more than 60 percent of patients suffer from a clinically evident neurological dysfunction. Neuropathologic changes are demonstrated in 75 - 90 percent of them at autopsy. HIV enters the CNS during the early phase of infection. HIV replicates predominantly in the nervous tissue macrophages which serve also as intrathecal reservoirs of infection. HIV isolated from the CNS is usually macrophagotropic. Neural cells are not susceptible to a productive HIV infection, contrasting with the permissivity of activated astroglial cells. The neuropathological picture of the brain involvement is typical by the giant multinuclear cells, i.e. fused monocytes/macrophages, then neuronal loss and changes in the white matter. The clinical manifestations of CNS involvement (AIDS encephalopathy) in HIV disease are variable protean, frequently associated with dementia. The pathogenesis of the neurological disease remains elusive. The cells supporting the HIV replication in the CNS (microglia, monocytes, astroglia) do not play a major role in dementia development. The neurotoxicity of viral glycoproteins, virus-induced cytokines and neurotoxin produced by CNS macrophages infected with particularly efficiently replicating HIV strains are being intensively studied. Dementia is associated with an increased virus load in the brain in the advanced stage of HIV disease. Neurotoxicity associated with HIV-infected microglial cells and macrophages activity remain to be considered, for the time being, as the most likely pathogenetic mechanism of neural dysfunction and injury. Our investigations have demonstrated that HIV infection of macrophages stimulate considerably the synthesis of MIP-1-alpha, MIP-1-beta RANTES chemokines (subgroup CC). These substances by their chemoattractant and activating properties may participate in the pathogenesis of HIV/AIDS encephalopathy, contributing to leukocytosis and inflammation, increasing thus the population of HIV-susceptible cells, facilitating their infection and enhancing finally the intrathecal spread of virus. (Tab. 2, Ref. 22.) PMID- 9333425 TI - [Autoimmune mechanisms in the pathogenesis of neurologic paraneoplastic diseases]. AB - Neurologic paraneoplastic syndromes (NPS) are disorders of the nervous system that are associated with the presence of malignant tumors. They are also known as the remote effects of cancer. There is mounting circumstantial evidence supporting the autoimmune origin of NPS. In NPS patients, paraneoplastic autoantibodies have been identified which react with the same or similar antigenic epitopes in tumor cells and neurons. However, except for the Lambert Eaton myasthenic syndrome (LEMS) it remains still unproven as to whether the autoantibodies are pathogenic or merely represent an epiphenomenon to the actual disease process, since the antigens identified by the paraneoplastic autoantibodies are primarily intracellular as to their location. Few data are available regarding the role of cell-mediated cytotoxicity and antigen dependent T-cell cytotoxicity. Further studies will be necessary to elucidate the role of autoimmunity in the production of paraneoplastic neurologic disorders. (Tab. 2, Ref. 6.) PMID- 9333427 TI - [Current issues in the development of the system for health protection of the population of Russia. (The reform: imitation or reality?)]. PMID- 9333426 TI - [Tirilazad mesylate (Freedox)--an effective inhibitor of lipid membrane peroxidation]. AB - The 21-aminosteroids (lazaroids) are inhibitors of lipid membrane peroxidation and appear to function as reactive free radical scavengers (RFRS). Freedox--a multimechanistic cytoprotective inhibitor of lipid peroxidation has been developed specifically to minimize secondary tissue damage. It is the first lazaroid compound used in clinical practice for critical care indications. Structurally described as a 21-aminosteroid, it has no glucocorticoid, mineralocorticoid, or other hormonal effects. Cytoprotective pathways of Freedox after its insertion into the lipid bi-layer of cell membrane include: scavenging reactive oxygen intermediates (ROI), stabilizing cell membrane by decreasing fluidity, preserving vitamin E content in membrane, increasing surface viscosity, preserving of post injury Ca+2 homeostasis. There was shown its efficacy in improving neurologic outcome following CNS trauma, subarachnoid hemorrhage, and ischemia. The therapeutic potential of the lazaroid Freedox has been extensively studied in several CNS disorders. There is an increasing experimental and clinical evidence about the oxygen free radical formation and cell membrane lipid peroxidation which play an important role in the pathogenesis of subarachnoid hemorrhage, spinal cord trauma, head injury and inflammatory processes of the NS. Freedox has also been tested in a variety of stroke models. (Fig. 1, Ref. 19.) PMID- 9333428 TI - [Opinions of chief physicians on the health care reform in the Novosibirsk oblast]. PMID- 9333429 TI - [The reforms of the health care system in the Samara oblast. Problems and prospects]. PMID- 9333430 TI - [Health care reform]. PMID- 9333431 TI - [Cardiovascular surgery in the Russian Federation: state of the art (1993-1994)]. AB - Analyzes the status of cardiovascular surgery in the Russian Federation in 1993 1994. Emphasizes that exhaustive utilization of its potentialities will have an appreciable impact of the health of the society and offers relevant recommendations. PMID- 9333432 TI - [From the new economic mechanism in public health to obligatory insurance]. PMID- 9333433 TI - [Considerations on the article by V.I. Shevskiy "Reforms of health care system in the Samara oblast: problems and perspectives"]. PMID- 9333434 TI - [Difficult paths of the reforms]. PMID- 9333435 TI - [Some aspects in financing public health institutions under the conditions of the obligatory medical insurance]. AB - Analyzes the data of sociologic study carried out among head physicians of therapeutic and prophylactic institutions of the Novosibirsk region, who were asked to express their opinions on the principal problems of public health and its reformation. Although the majority of respondents consider the reforms necessary, almost half of them are sure that the direction of reforms is wrong. PMID- 9333436 TI - [Problems in prophylaxis under the conditions of public health reform]. AB - The indicators of medicosocial significance of prophylaxis in present-day Russia have been determined on the basis of published data and authors' findings. The weak and strong aspects of the problem are assessed, as are the potentialities of solving it and factors threatening its realization. Problems in the prophylactic activities of general practitioners (family physicians) and problems in the development of prophylaxis within the framework of medical insurance are discussed. PMID- 9333437 TI - [Methodological approaches to the evaluation of medical-economic standards and possibilities of their utilization in health care]. AB - Medicoeconomic standards (MES) became very popular in many regions of Russia as a unit for assessing the monetary relationships between hospitals and insurance companies within the framework of obligatory medical insurance. However, rather often the standards are introduced by voluntary solutions without due consideration for the specific features of a territory, this involving the risk of a formalistic approach to the use of MES. Regulations have been designed for the experiment on comparison of the medical standards which have been approved and the actual treatment technology in the hospitals of Moscow (actual volumes, bed-days, and cost of treatment). Such an experiment will help assess the current MES as far as it concerns their correspondence to practical medical care and will be conducive to their improvement and to the creation of new models of MES. PMID- 9333438 TI - [Social medical insurance and public health management in Austria]. PMID- 9333439 TI - [First professors of the medical department at the Moscow University]. PMID- 9333440 TI - [The history of international relations of the People's Committee on Public Health in the 30th (For the 100th anniversary of the birth of G.N. Kaminskogo++]. PMID- 9333441 TI - [Health and life style of families among various social groups of the population with young children]. AB - Presents the results of a sociohygienic study of the health status, life style and conditions of 900 families belonging to different social groups (workers, employees, and mixed) with young children. The principal risk factors for children's health are chronic diseases and adverse occupational factors of their parents, poor microclimate in the family, and poor education of their mothers. Medicosocial requirements of workers' families are a priority problem. PMID- 9333443 TI - [The medal "Iron and Blood". (On the 140th anniversary of the end of the Crimean War)]. PMID- 9333442 TI - [The medical professionals of the underground in the Sumy region in the years of the Great Patriotic War]. PMID- 9333444 TI - [Dynamics of abortion in various regions of Russian Federation]. AB - Analyzes statistics of induced abortions as reflected at the Ministry of Health and Medical Industry of Russia. Puts forward the most significant medicosocial factors influencing the causes of pregnancy discontinuation. Analysis of the prevalence of induced abortions in different regions of Russia persuasively demonstrates the necessity of improving the state statistics of abortions for the realization of family planning programs. PMID- 9333445 TI - Optimal needle lengths determined for deltoid immunization in adults. PMID- 9333446 TI - Diabetes Hypertension Angiopathies. Proceedings of the 15th International Karlsburg Symposium on Problems of Diabetes. Karlsburg, Germany, September 8-11, 1995. PMID- 9333447 TI - [Age adjusted diagnosis/normal values in geriatrics--sense and nonsense]. PMID- 9333448 TI - [Homeostasis and adaptability in the elderly]. PMID- 9333449 TI - [Reference ranges in geriatrics: a review of age-dependence of selected blood components]. AB - An age dependency of blood components is well documented especially in childhood. Less investigated is the situation in the elderly population. Study of the literature and our own investigations show that concentrations of several analytes remain constant throughout adulthood, but many of them increase or decrease with age. The interindividual variability of most parameters is higher in elderly people than in younger adults. Therefore, it is not recommended at the present time to use special adapted reference intervals, because the larger variation of values in higher ages could be affected by the inclusion of diseased individuals in the reference population. Also in high ages, where biochemical individuality is largely maintained, intraindividual reference ranges should be preferred. Thus, latent diseases could be detected much earlier than by transversal evaluation of data. PMID- 9333450 TI - [Cardiopulmonary normal values in geriatrics]. AB - Because of age-related modifications of the heart, lung, and circulation, several cardiopulmonary parameters present altered normal and reference values with increasing age. Other parameters, for example, arterial blood pressure, change only insignificantly with age. In older age groups it is more difficult to establish reference ranges and values because of the lack of "healthy" subjects. To establish cardiopulmonary parameters, appropriate investigations for the age group, for example, treadmill-ergometer, should be preferred. In judging if a value resulting out of the reference range is to be considered really pathologic, other factors are to be taken into account, such as circadian variations, motivation, associated pathologies as well as the actual physical and psychological conditions, when we are dealing with older subjects or patients. PMID- 9333451 TI - [The kidneys in aging]. AB - A progressive deterioration of renal anatomy and renal function with aging is inevitable. Glomeruli become sclerotic, renal mass declines, kidneys are shrinking and creatinine clearances decreases about 50% after the third decade of life. Renal aging in accelerated by risk factors of arteriosclerosis. The relationship of serum creatinine and creatinine clearance changes with age, since creatinine production is diminished. Old kidneys show a reduced ability to compensate stress, e.g., infection, medication, changes in blood pressure, and the risk of acute renal failure is increased. A glomerulonephritis in advanced age may be caused by a vasculitis; despite rapidly progressive course, early treatment can often improve renal function. The dosage of drugs eliminated by the kidneys has to be adapted in old patients. Despite only moderately elevated serum creatinine, renal function may be severely reduced in old persons and dialysis may be indicated. Dialysis and renal transplantation are possible therapeutic options despite advanced age. PMID- 9333452 TI - [Gastrointestinal problems in elderly patients]. AB - The following article contains a short review on gastrointestinal problems of the elderly. The diseases of the esophagus occurring in the elderly are not much different from those in younger patients. Clinically relevant in the stomach are above all bleeding ulcerations and the gastric carcinoma occurring more frequently in advanced age. The pyogenic liver abscess is diagnosed primarily in the elderly and is at a rule the consequence of an infection of the gall bladder and other abdominal sites. The hepatocellular carcinoma does not grow rapidly in the elderly, but its accompanying unfavourable survival rate at five years is also approximately 5 per cent. In the case of symptomatic cholelithiasis, older high risk patients do especially profit from minimally invasive laparoscopic surgical procedures. Today, bile duct calculi are preferably treated by endoscopic papillotomy and following extraction of the calculi. The pancreas is subjected to atrophy, lipomatosis and fibrosis at the advanced age. However, these changes are rarely of clinical relevance. A frequent problem in clinical practice is that of constipation, from which 35% of patients suffer above the age of 65 years. Another typical symptom of the elderly is the incontinence, the different causes are being discussed. In advanced age, gastrointestinal hemorrhages are mostly occurring above the Treitz's ligament. Hemorrhages of the lower gastrointestinal tract occur mostly in the form of diverticle bleedings and those of angiodysplasias in the elderly. The diverticulosis is also a disease observed in over 50 per cent of patients above 70 years, but it is symptomatic in only part of the patients. When suspecting an inflammatory bowel disease in the elderly, the possibility of a mesenterial ischemia must always be considered as differential diagnosis. The classical chronic inflammatory bowel diseases can, however, also occur at advanced age. The colon carcinoma is one of the most frequent lethal causes in the Western countries 90 per cent of the cases of colon carcinoma are found in patients older than 50 years of age. Intensive attention is therefore required in this age group. PMID- 9333453 TI - [Characteristics of immunologic test values in the elderly]. AB - The immune system changes during the lifespan of man. Many described changes in the immune system of the elderly were dependent on illness or chronic diseases. To exclude these pathological changes in the immune system and to exclusively describe age-dependent changes, Ligthart et al. defined immunogerontological criteria to study the immune system in the elderly, the SENIEUR-Protocol. Most changes in the immune system of elderly are within the normal ranges of the appropriate parameter. However, there are many significant differences between the status of the immune system in healthy young and elderly individuals, within these normal ranges. The comparison between SENIEUR-elderly and healthy young and the additional comparison of these two groups with centenarians allows the discussion of potential pathological effects of these changes. In this article we summarize the described changes of the immune system in SENIEUR-elderly and centenarians. The serum levels of the immunoglobulins G, M and A increased with age, as well as the number of benign monoclonal gammopathies and the number of autoantibodies. The titers of zinc are significantly decreased in the serum of the elderly. The production of the acute phase protein C-reactive protein is not age-dependent, whereas the serum levels of alpha 2-macroglobulin are significantly increased in the elderly. The number of lymphocytes decreased and the number of neutrophils increased with aging. Monocytes, basophils, and eosinophils are without changes during life. There are many descriptions about changes of the leukocyte sub-population in aging, which are not always comparable. However, the number of T cells (CD3) decreases. Within the T cells the CD8 cells decreased more than the CD4 cells, resulting in an increased CD4/CD8 ratio. Memory T cells (CD45RO) increase during life, whereas naive T cells (CD45RA) decrease. Interestingly, centenarians have more naive T cells SENIEUR-elderly. The number of B cells (CD19) decreased also, whereas the number of natural killer (NK) cells (CD16, CD56, CD57) increases with aging. The capacity of leukocytes from the elderly to produce cytokines is also significantly different from those of the young. The release of the TH1-cytokines interleukin (IL)-2 and interferon (IFN)-gamma is decreased, whereas the production of the TH2-cytokines IL-4 and IL-10 is increased in the elderly. The production of proinflammatory cytokines such as IL-1, IL-6, IL-8, and tumor necrosis factor-alpha is increased in the elderly. In contrast, the capacity to produce the antiviral cytokine IFN-alpha is reduced in elderly individuals. In conclusion, the immune system shows many age-dependent changes, but we know little about the reason and the potential pathological effects of these changes. PMID- 9333454 TI - [Radiology and geriatrics: limits between normal values and pathology exemplified by conventional thoracic roentgen image, cerebral CT and MRI and pelvic-leg angiography]. AB - Radiological examinations and intervention in the elderly are a matter of routine but require special consideration by technical and medical staff because of specific problems caused by the morbidity of the patient. Aging and degenerative processes of the organism have to be included in the interpretation of the findings. Discriminating pathological from age-related alterations is occasionally difficult. This paper demonstrates senile pulmonary emphysema, physiological brain atrophy and arterial angiosclerosis as typical examples for age involution. Age-related changes are compared with similar pathological findings. Clinical symptoms of the patient are decisive for the interpretation and classification of images as well as the subsequent therapeutical procedure. PMID- 9333455 TI - [Geriatric assessment--the Vienna model]. AB - Geriatric Assessment is a structured interdisciplinary process for the diagnosis of the functional status and potential for rehabilitation of geriatric inpatients. This paper describes the possibilities and scope of the Vienna Model of Geriatric Assessment oriented in terms of practicability and focused implementation of resources. By using this multidimensional proceeding the rehabilitation-quotient of patients has increased more than 39%. From this point of view Geriatric Assessment in a geriatric hospital is a practical instrument to prevent hospitalization, to increase survival at home, and to improve functional status in elderly patients. PMID- 9333456 TI - [A bio-psychosocial treatment concept improves coping with illness by geriatric patients]. AB - 37 geriatric patients, who had been treated in hospital on the base of the biopsychosocial model of illness, were compared with a control group of 36 patients, who underwent traditional therapy of internal medicine, in regard to coping with illness. Both samples did not differ in diagnosis and other characteristics of the illnesses, e. g., social situation, institutional factors and ways of hospital organisation and coping styles. But as suspected the patients treated with the biopsychosocial concept felt better supported by the medical staff and were more content with treatment, and the mental adaptation to illness was better before discharge from the hospital. We conclude that the "therapeutic environment" using a biopsychosocial concept of treatment helps geriatric patients to cope successfully with illness. PMID- 9333458 TI - [The use of plasmosorption on SKN IK sorbent in a severe course of leptospirosis]. AB - In severe course of icterohemorrhagic form leptospirosis presenting with an acute renal-hepatic insufficiency and manifest hypocoagulation (BC II-III), plasmasorption on the sorbent SKN [correction of CKH] IK with carboperfusion of 0.8-1.2 1 plasma appears to be warranted. Being endowed with an apparent detoxicating effect at the expense of absorbtion of creatinin, urea bilirubin, this manipulation, with small doses of heparin, promotes also stabilization of hemostasis and arresting of hypocoagulation, which fact enhances appreciably the compensatory power of the organism improving the prognosis of the condition. PMID- 9333457 TI - [A trial of the use of the multivitamin Multitabs in the combined treatment of patients with a perinatal brain lesion]. AB - Pediatric patients in early childhood presenting with perinatal affection of the nervous system benefit much from incorporation into their combined treatment of multivitamins "Multitabs", as evidenced by improvement in their general health, as well as in the red blood parameters and immunity status. Thus, use of the above multivitamins for children presenting with perinatal cerebral pathology is considered liable to be of benefit making for optimization of the process of treatment. PMID- 9333459 TI - [Sweet's syndrome]. AB - Kept under observation were eight patients with Sweet's syndrome--six men and 2 women aged 25-60. The syndrome was accompanied by fever in 8 patients; in 6 it was preceded by common cold, respiratory diseases; neutrophilia was present in 7 patients. Rash presented as inflammatory spots, 10 am in diameter, was located predominantly in the upper part of the body. Helpful in the treatment of the condition were corticosteroid therapy and potassium iodide. PMID- 9333461 TI - [Cardiac contractile function in the participants of the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station who are ill with hypertension]. AB - In a major proportion (67%) of those examined (persons with stage-II hypertensive disease who took part in the elimination of the aftermath of the Chernobyl breakdown) with the aid of echocardiography, hypertrophy of the left ventricular (LV myocardium was revealed as was augmentation of the mass of the LV myocardium together with decline in the contractile function, while in other examinees (33%) all above parameters showed no deviation from the norm. This permits predicting probability of cardial complications in such patient populations. PMID- 9333460 TI - [New approaches to the therapy of hypercalcemia in cancer]. AB - An overview of the published literature provides current information about pathogenesis and treatment of hypercalcemia, and focuses on its treatment with a new class medications, such as biphosphonates (ethidronate, clodronate and pamidronate). Use of these drugs permits reducing complications caused by metastases of malignant tumors to bones. PMID- 9333462 TI - [Bottled Zbruch naftusia mineral water--a new immunocorrective for regions ecologically at crisis]. AB - Effects were studied of bottled mineral water "Zbruch Naftusia" on the blood and immunity condition of hypothetically healthy population of the Donetsk region living under environmental hazard conditions and having disorders in the area of the immune system. The studies made showed that prescription of the above drug preparation for use by conditionally healthy population of the Donetsk region, just those very contingents maintaining long-time occupational contact with liquidators of aftermath of the Chernobyl breakdown, is associated with immunocorrection at the level of content of the immunocompetent cell populations and subpopulations, with their function restored against the background of alleviating of the syndrome of endogenous intoxication that had developed because of them being resident under health hazard conditions in an ecocrisis-ridden region. PMID- 9333463 TI - [The embryotoxic and teratogenic action of some brands of epoxy resins]. AB - The studies made permit stating that such epoxy resins as DE-500 with dose levels of 1/10 and 1/50 DL50, DE-1000 with 1/10, 1/50, 1/100, 1/250 DL50, UP-650, UP 650T with 1/10 DL50 administered into the stomach of pregnant females of nondescript albino rats from the 1st to the 19th day of pregnancy (DL50 being 5338 +/- 1134, 6644 +/- 1114, 9180 +/- 1154, 7717 +/- 586, 6980 +/- 621 mg/kg for the resins DE-500, DE-1000, DE-2000, UP-650, UP-650T respectively) have embryotoxic as well as teratogenic effects. The embryotoxic effect of the above epoxy resins depends on the dose administered and their chemical structure. These same embryotoxic and teratogenic effects a need to be taken into consideration in scientific substantiation/revision of hygienic regulations for polyoxipropilene diepoxides and alicyclic epoxy resins. PMID- 9333464 TI - [English walnuts in the combined treatment of rheumatoid arthritis patients]. PMID- 9333465 TI - [Endotoxicosis in the pathogenesis of severe forms of psoriasis and its correction by combined therapy using sillard P and plant species]. PMID- 9333466 TI - [The evaluation of the continuity status in the delivery of medical care to therapeutic patients in home hospitals]. PMID- 9333467 TI - [An approach to the structuring of printed materials in education technology for the independent study of the pharmacotherapy of emergency states]. AB - In view of the fact that the sources of information (text-books, manuals, monographs) used in higher medical institutions lack a clear-cut system of directing the learning activities of those being thought, we have devised a variant of a manual that stipulates that a goal be defined, the content of pharmacotherapy of urgent states chosen, teaching material organized, a system of individual tasks set, results of learning controlled, auto-education and self control exercised. The above type manuals are supposed to be a means of directing the activity aimed at learning to apprehend and must be a mainstay in the realization of the programme-deliberate management of quality of training a specialist. PMID- 9333468 TI - [Cancer from an ulcer or ulcerative stomach cancer? (a review of the literature and the authors' own observations)]. AB - On the basis of surveys of literature and the author's own experience of work, ulcerous lesions of the stomach should be regarded at present as a facultative (precancerous) condition rather than obligate precancer. A great many of biopsy specimens need to be taken from the patient's stomach across the lifespan and subjected to morphologic study on a repeated basis before an adequate answer can be given to a question of a possibility of developing cancer from chronic ulcer of the stomach. PMID- 9333469 TI - [The classification of the initial forms of cerebral vascular diseases]. PMID- 9333470 TI - [The history of the first joint scientific and practical measures of the Ukrainian Institute of Physician Advanced Training and of the public health institutions of Sumy Province]. PMID- 9333471 TI - [The Kiev Research Institute of Epidemiology and Infectious Diseases (on the centenary of its founding)]. PMID- 9333472 TI - [Monuments to physicians for their heroic deeds in wars]. PMID- 9333473 TI - [The current concept of early postinfarct stenocardia and the procedure for patient management]. PMID- 9333474 TI - [The new nonsteroidal anti-inflammatory preparation meloxicam in the treatment of rheumatoid arthritis]. PMID- 9333476 TI - [The basic tenets of the new concept of balanced nutrition]. PMID- 9333475 TI - [Disordered visual functions in patients with a traumatic carotid-cavernous fistula]. PMID- 9333477 TI - [The immunological characteristics of systemic vasculitis]. AB - A total of 142 patients with systemic vasculitis (SV) during the phase of activation of the process were evaluated. Diagnosis was based on the clinical as well as laboratory, functional and morphologic findings. Immunological investigation revealed increase in T-lymphocyte effector activity, marked sensibilization of lymphocytes to the intimal antigen, hyperimmunocomplexemia, decline in phagocytosis, hypocomplementemia. Musculocutaneous biopsies showed fixation of IgG and the complement C3-component. For different SV variants agreed specific markers were determined. The results obtained permit objectivizing different immunotherapeutic alternatives in SV treatment. PMID- 9333478 TI - [A comparative analysis of the rheographic indices of the pancreatic blood flow in patients who have undergone a subtotal gastric resection and gastrectomy]. AB - Overall ninety-two patients with carcinoma of the stomach were studied, 45 of whom had undergone subtotal resection, and 47--gastrectomy. During the first postoperative 10 days a study was made of the pancreatic hemocirculation by contact bipolar rheography. After surgery on the stomach, the pancreas shows phasic changes in bloodflow that include the time of arterial hyperemia, that of insufficiency of arterial blood-filling, and time period of restoration of bloodflow. Fluctuations in pancreatic arterial blood filling in those patients who had survived gastrectomy within the first 48 hours following the operation are more definite than in those having undergone subtotal resection of the stomach. PMID- 9333479 TI - [The blood concentration of flurenizid in patients with destructive pulmonary tuberculosis]. AB - Some pharmacokinetic features of new capsulated and tableted antituberculous drug flurenizidum were studied microbiologically in 50 adult patients with advanced pulmonary tuberculosis. Flurenizidum in vitro shows high mycobacteriostatic activity: it suppresses growth of Mycobacteria tuberculosis H37Rv strain in concentration of 0.05 mkg/ml. Single doze of flurenizidum--0.3 and 0.6 g- provided mycobacteriostatic blood concentration for 6 and 12 hours respectively. Maximum concentrations of the drug (0.46 +/- 0.07) mkg/ml and (1.02 +/- 0.14) mkg/ml respectively were observed in 3 hours after administration. PMID- 9333481 TI - [Duodenogastric reflux in patients with alimentary obesity]. AB - The authors analyze findings from a comprehensive clinical and instrumental evaluation of 59 patients presenting with duodenogastric reflux. The above findings suggest that underlying the duodenogastric reflux in those patients having surplus body mass might be hypotonia of the stomach, particularly its pyloric portion. In patients with body mass that do not deviate from physiologic norms regurgitation of bile into the stomach results from the presence of duodenal dyskinesia by hypertonic type. The results obtained suggest that patients with gastroduodenal reflux need treatment options for them to be carefully thought out. The above reflux appeared to be particularly pronounced in those patients presenting with alimentary obesity as per analysis of the gastric content. Patients with grade II obesity show more noticeable tonic disorders of the pyloric portion of the stomach. PMID- 9333480 TI - [Sympathoadrenal activity during the treatment of patients with cancer of the stomach and large intestine]. AB - Excretion was studied of catecholamines and diphenilamine (DOPA) in 310 patients with carcinoma of the stomach and large intestine and 43 patients with non malignant diseases. The oncological patients showed decrease in activity of the mediator link of the sympathoadrenal system (SAS) as well as its reserves but there was no association with sex, age, location or histological structure. Three types of SAS functioning were identified, such as compensation, overstrain and emaciation. Surgical intervention led to activation of the hormonal link and exhaustion of the system's reserves. Two kinds of sympathoadrenal response to stress were described--adequate and inadequate. In the former type, phases of stress remain as they are, unchanged, as are time periods of formation thereof, while under the latter one time periods of the phases formation or formation thereof get disordered. PMID- 9333482 TI - [The hemodynamic and functional effects of using embolization of the splenic artery and endoscopic sclerotherapy in patients with liver cirrhosis at the decompensation stage]. AB - Complex studies were made of hepatoportal hemodynamics and functional reserves of the liver in 30 patients with decompensated cirrhosis following combined use of embolization of the spleen artery and sclerotherapy of varicose veins of the esophagus. Clinical assessment of the results obtained revealed stabilizing effect the sparing interventions have on the course of the underlying pathological process. PMID- 9333484 TI - [The immunological characteristics of patients with chronic bronchitis and a concomitant drug allergy]. PMID- 9333483 TI - [Transvesical partial electric resection of prostatic adenoma in patients with a high degree of surgical risk]. PMID- 9333485 TI - [The correction of the status of the pulmonary surfactant system as a method for preventing experimental pneumoconiosis]. AB - Using a model of experimental pneumoconiosis in albino rats, induced by breathing in a rock dust, we attempted correction of the pulmonary surfactant system, for which purpose we tried choline chloride as a drug preparation. The action of the above drug was found out to be depended upon the moment of its administration and time during which it is being administered, and is that of influence on both the concentration and mechanism of absorbtion of surface-active fractions of the surfactant in the liquid-gas interface. Early administration of choline chloride (during the dust factor exposure) increases concentration of those surfactants notable for their surface-active properties being maintained, and improves the processes of diffusion of surface-active substances, which observation is accompanied by decrease in intensity of the processes of fibroformation in the lung tissue. Choline chlorid employed on the discontinuation of the dust exposure has adverse effects leading to changes in the lipid composition, impairing the surface-active properties and processes of diffusion of the pulmonary surfactant, which fact may contribute to aggravation of fibrosis in the lungs. PMID- 9333486 TI - [The comparative characteristics of the clinico-hematological indices in patients with different forms of myelodysplasias]. AB - Clinical findings were analyzed as were parameters of the peripheral blood and bone marrow in 102 patients presenting with hematodysplasias with regard to a form of the disorder and in comparison with healthy subjects. Used for the analysis was the computer system "GEMA" (Saint-Petersburg). Correlations were analyzed between the studied parameters. Differential-diagnostic differences were revealed between refractory anemia, blast surplus refractory anemia, and refractory anemia with blast surplus in the phase of transformation into acute leukosis. The results obtained can be used for their differential diagnosis, prediction of the disease course, choice of therapeutic alternatives. PMID- 9333487 TI - [The chronorhythms of kidney functions in diabetes mellitus]. PMID- 9333488 TI - [The cerebral hemodynamics in subjects with the initial manifestations of insufficiency of the cerebral blood supply]. PMID- 9333489 TI - [The formation of atherosclerotic prodromes in risk-group adolescents at stages in their sexual maturation]. AB - 240 adolescents with primary arterial hypertension and 118 youngsters from families with aggravated heredity in respect of ischemic heart disease were studied for lipidic spectrum of blood, system of prostanoides and some hemodynamic indexes during the puberty stages. It has been found out that adolescents of risk groups for atherosclerosis develop metabolic disturbances presented as atherogenic dyslipoproteinemias and thrombogenic balance of prostanoides. A particular feature of the above disturbances is their apparent stability, they are responsible for structural alterations in the vascular wall presented as its thickening and loss of elasticity. This suggests that not only metabolic basis of atherosclerosis might be formed during the pubescence stages but structural one as well. PMID- 9333490 TI - [A comparative study of the hemodynamic and antioxidant effects of Capoten and prazosin in patients with refractory heart failure]. AB - Overall fifty one patients with chronic cardiac insufficiency (ChCI) were studied for changes in parameters characterizing hemodynamics and lipid peroxidation (LPO) under treatment with kapoten and prazosin. Kapoten was found to be capable of exerting an antioxidant effect and working in ways beneficial for the pulmonary circulation, while prazosin is generally indicated to patients with ChCI presenting with increased end diastolic pressure in the left ventricle. Prazosin activates LPO, for which reason its pro-oxidant action needs to be drug corrected. Because of marked pharmacologic effects of both drugs in dealing with refractory circulatory insufficiency it is advisable that further studies be made in order that we might be able to determine indications for kapoten and prazosin therapy in coronary patients as well as those with arterial hypertension, cardiomyopathies, valvular defects, ChCI with complications more accurately. PMID- 9333492 TI - [The personality characteristics of patients with duodenal peptic ulcer]. AB - Using MMPI test, 196 patients with duodenal ulcer were studied for their personal traits. The patients' personality was characterized by a high level of anxiety, mental tension, emotional immaturity, affective rigidity, suppressed aggression. The most apparent in the young adults were deviations from the norm of psychopathic nature; as patients grew older, their illness of longer duration, their personal profile tended to be dominated by a problem of suppressed aggression and emotional immaturity, with hypochondriac and depressive tendencies to be on the increase. The personal traits did not appear to be dependent upon severity of the disease course or much affected by the treatments administered that incorporated tranquilizers, antidepressants and a novel synthetic opioid hexapeptide dalargin. PMID- 9333491 TI - [The immunomodulating effect of vilozen in the combined treatment of children with rheumatism associated with an allergy]. AB - Effects were traced out of vilosene alone and combination with ketotifene on clinical-and-immunologic parameters in 86 children with rheumatism associated with concurrent allergy. There has been disclosed immunomodulating effect of vilosene in hyposuppressor variant of a secondary immune deficient state in rheumatic children. Antiallergic action of vilosene is more pronounced when employed in combination with ketotifene. The clinical effect was evident 1-1.5 weeks earlier than in controls. The data obtained suggest vilosene and ketotifen to be useful in the treatment of rheumatic children presenting with concurrent allergy. PMID- 9333493 TI - [A new theoretical and experimental validation of the treatment and prevention of cholelithiasis]. AB - Approaches are proposed, new in principle, toward breaking biliary calculi into pieces, as well as toward prophylaxis of cholelithiasis. Exposure of the concrements to ultrasound with varying resonance frequency in a contact medium containing litholytic substances permits bringing about their reduction to fragments being safe for their evacuation from the biliary system. Prevention of cholelithiasis involves identification of precalculous period and exposure of the gallbladder region to vibration along with adopting conventional therapy. PMID- 9333494 TI - [The intra-arterial chemotherapy of tumorous lesions of the stomach, rectum and liver]. AB - Roentgenosurgical treatment of malignant tumor of the stomach, straight intestine, liver is based on a directed bringing of cytostatics to the tumor by way of catheterization of afferent vessels together with the use of the modifier of drug resistance verapamil. A higher therapeutic effect can be achieved by securing an easier access of the medicinal substances to the membrane of the tumor cells. There can be noted regression and destruction of the tumor, enhancement of ablasticity of surgical intervention, with metastatic spreading and local relapses developing at a lower rate, which facts taken together contribute to a better outlook for the patients' survival and quality of their life. PMID- 9333495 TI - [The irritable bowel syndrome. The prospects for improving the quality of life]. AB - The lecture focuses on current concepts of etiology and pathogenesis of irritable colon syndrome and deals extensively with clinical manifestations of symptoms of the disease. Recommendation are given as to diagnosis and choice of optimal approaches to therapy with special reference to prospects for improvement of quality of the patients' life. PMID- 9333496 TI - [The characteristics of patients hospitalized in a general therapy department in connection with emergency pulmonary states]. AB - An analysis was done of urgent pulmonologic states, because of which states those hospitalized had been sent to a general therapy department. Over the year, 237 persons had been admitted into hospital through emergency first aid service, with urgent pulmonologic states making up 51.5% (n = 122) of the total, bronchial asthma (BA) and acute lung inflammation (ALI) predominating (55 and 55 subjects respectively). BA patients were found to be unable to recognize the worsening of their state in good time, they fail to keep in touch with pulmonologists and district hospitals. The main reason why ALI patients had been admitted into hospital on an urgent basis was intoxication syndrome and broncho-obstructive syndrome. In 20% of ALI hospitalizations, there was divergence in diagnoses (referral versus clinical ones). PMID- 9333497 TI - [The use of the Multitabs vitamin preparation in men with infertility]. PMID- 9333498 TI - [Kidney functional status in patients with staghorn nephrolithiasis complicated by chronic kidney failure after an organ-preserving surgical intervention]. AB - On the basis of examination of 125 patients with coral nephrolithiasis complicated by chronic renal insufficiency, who underwent various organ-sparing surgical interventions, the author has come to a conclusion that functional capacity of both the operated and the contralateral kidney gets impaired after the operation, its degree being directly related to the particular stage of chronic renal insufficiency. Adoption of an adequate antibacterial and disintoxication therapy in combination with hemodialysis (if indicated) in the postoperative period permit improving not only the functional state of the two kidneys but also the outcome of the organ-sparing surgical intervention. PMID- 9333499 TI - [The clinical efficacy of tanakan in patients with stage-I atherosclerotic circulatory encephalopathy]. PMID- 9333500 TI - Proceedings of the 7th L.H. Gray Workshop. Research Strategies for the Prevention of Early Events in Human Radiation Carcinogenesis. Dublin, Ireland, December 5-8, 1996. PMID- 9333501 TI - [The safety and usefulness of transgenic plants]. AB - Transgenic crop plants, used in food and feed production, carry different beneficial transgenes, mostly for resistance to pests, herbicides and diseases. All new transgenic plant varieties, the genes they carry and their products have been thoroughly tested before released for agriculture and even more for marketing. Genetically modified organisms carry the same risk as any other organism. Food derived from genetically modified organisms due to legal regulation is most controlled and therefore most safe food ever placed on the market. In future, transgenic plants offer many new possibilities also for medical use, like plant vaccines, antibiotics and rare proteins of clinical importance produced by plants. PMID- 9333502 TI - [Reflux disease]. AB - Reflux disease is a very frequent condition; its most frequent symptom, pyrosis, is found daily in 7% of the population. The disease is dut to the aggressive action of the gastroesophageal reflux "faced" by defensive oesophageal mechanisms. Reflux disease is classified as endoscopically positive with an obvious finding of oesophagitis and endoscopically negative with a normal endoscopic finding but detectable histological changes. Complications of the disease include strictures, ulcers, haemorrhage and Barrett's oesophagus. In the therapeutic regime provisions are used, medicamentous treatment (antacids, sucralphate, prokinetics, antagonists of H2 receptors and proton pump inhibitors) and surgical treatment. In the therapeutic strategy two procedures are possible: 1. the method of choice (treatment with a single drug) and 2. stepwise therapy a) step-up-the basis are regime provisions and gradually drugs are added, b) step down-treatment is started with the most effective drug, the proton pump inhibitor, and is reduced gradually). Treatment has a short-term character (treatment of the acute attack) and long-term (maintenance treatment). Contemporary therapeutic possibilities improve the prognosis of the disease and the quality of life of patients with reflux disease to the quality level of the normal population. PMID- 9333503 TI - [The postmenopausal estrogen deficiency syndrome and hormone replacement therapy]. AB - The oestrogen deficiency syndrome comprises all subjective and objective complaints and all organic and metabolic organ damage encountered after natural and induced menopause. In the development of these pathological conditions participates in a marked way endogenous oestrogen deficiency - oestrogen deficiency. The oestrogen deficiency syndrome affects some topographically remote organs and is manifested by an apparently clinically heterogeneous symptomatology. The clinical picture of the syndrome can therefore be very varied. It affects the mucosal membranes of the vagina, urethra, trigonum of the urinary bladder, oral cavity, nose, pharynx and larynx, eyes and large intestine, as well as the skin and its adnexa, the breasts, skeleton and cardiovascular system. The syndrome involves therefore all medical disciplines. The common origin of all pathological conditions ensuing from oestrogen deficiency calls for interdisciplinary collaboration in a new discipline - climacteric medicine. Exogenous oestrogens in different forms hold an important place in the prevention and treatment of oestrogen deficiency syndrome. In the first place it is prevention and treatment of arteriosclerosis, ischaemic heart disease, cerebrovascular attacks and osteoporosis. If we add undoubtedly favourable effects of hormonal substitution therapy on vasomotor complaints and symptoms of the organ syndrome, then the advantages of HRT markedly overrule its risks. PMID- 9333504 TI - [Compliance in treatment of the sleep apnea syndrome using continuous positive pressure respiration]. AB - BACKGROUND: Continuous positive airway pressure (CPAP) is the therapeutic method of choice in sleep apnoea syndrome (SAS) but involves at first discomfort for the patients. The correct indication, correct setting of the overpressure and good adaptation influence the application and therapeutic asset of CPAP. METHODS AND RESULTS: From a total number of 41 patients with SAS treated by CPAP 24 were examined (incl. three women), who had CPAP in domiciliary treatment for more than two months. The mean period of use was 288.2 days (range 52-824). 84% of the patients used CPAP daily, the mean number of applications per week was 6.26 (range 2-7). For the whole period of sleep CPAP was used by 75% patients and the mean sleep period with CPAP was 6 hours (range 2.5-8). None of the patients discontinued treatment completely. Four patients used CPAP inadequately (less than 25 hours per week) - one because of intolerance associated with severe CHOCHB, the second one for intolerance of overpressure of 15 mbar, the third one because of poor motivation and the fourth one because of dehydration of mucous membranes. All patients recorded the therapeutic effect of CPAP. Rhonchopathy disappeared in 87.5% patients, excessive somnolence improved in 91.7%, fatigue declined in 88.5%. The functional capacity and work performance improved in 95.8% patients. Undesirable effects were not serious nor frequent: escape of air from the mask (29.2%), dry mucosae (20.8%), pressure sores caused by the mask (20.8%), serous rhinitis (12.5%), burning sensation of the mucosae of the upper airways (8.3%) and conjunctivitis (4.2%) CONCLUSIONS: Adequate compliance with CPAP was recorded in 83.3% patients and a favourable effect of treatment was proved. PMID- 9333505 TI - [Present views on boxing--Henner's legacy]. AB - Fifty years ago academician Henner formulated clearly the negative view of neurologists on boxing as practised in his time. It was his merit that a protective helmet was introduced, many of his demands concerning the regime after a KO and the importance of medical examinations were no yet adequately appreciated. At the opportunity of the 100th anniversary of Henner's birth and the fiftieth anniversary of the Clinic he founded in the premises of saint Catherine the author discusses contemporary views on boxing, the risk of repeated injuries for the development of encephalopathia pugilistica and their relationship to Alzheimer's disease. Contemporary boxing is a spectacle which is a continuation of historical fights of gladiators. If the main objective is to hurt the adversary, this activity does not deserve the name of "sport" Sport should promote and maintain physical and mental health which is not the case in professional boxing. PMID- 9333506 TI - [Trace elements, human nutrition and health]. AB - The importance of trace elements as essential dietary constituents is not sufficiently appreciated by the medical profession, although in recent years a significant increase of new finding on their essential character in nutrition occurred as well as of data on changes in the composition of the diet which can cause their deficiency and in some instances a toxic excess. The diagnosis of deficiency or excess of these substances in the organism is very complicated. because their blood level frequently does not provide sufficient information on their reserves in the organism and new laboratory methods suited for this purpose must be developed and tested. The interpretation of the assembled results is very pretentious and it will be therefore necessary to expand teaching of nutrition, where the problem of trace elements no doubt belongs, into undergraduate and postgraduate medical curricula to prevent health damage due to inadequate intake of trace elements as well as uncontrolled intake of freely available preparations sold as supplements. PMID- 9333507 TI - [The physiology of erection]. AB - The majority of contemporary knowledge on the physiology of erection was assembled during the past thirty years. Today we consider erection as a multifactorial process. Mechanically it can be compared to an electromechanically controlled hydraulic system. Its function is conditioned by a number of mutually coordinated processes. As to nervous processes they include autonomous (parasympathetic and sympathetic) innervation, as well as somatic innervation (sensory and motor pathways). The control function is exerted by spinal as well as cerebral centres. As to mediators, in particular acetylcholine, nitrous oxide (NO) released from the endothelium are involved, noradrenaline, VIP (vasoactive intestinal polypeptide), CGRP (calcitonin gene-related peptide) and prostaglandins. The most important roles in the phase of erection are played by nitrous oxide and VIP. Erection can be either reflex erection, psychogenic or nocturnal or morning. It usually takes place in six stages (at rest, latent, the tumescence stage, complete erection, rigid erection and subsequently the stage of detumescence). Except for neurohumoral mechanisms an essential prerequisite for the development of erection are the arterial supply of the genital and the so called venoocclusive mechanism. Erection takes the following course (simplified): erotogenic stimuli lead to the stimulation of the parasympathetic nerve- >vasodilating substances are released-->the s inusoids are filled with blood (tumescence stage)-->the venoocclusive mechanism starts to work: thus complete erection occurs. Then the contractions of the musculature of the perineum compress the proximal portions of the corpora cavernosa: this leads to rigid erection. Detumescence which occurs as a rule after ejaculation) is due to released noradrenaline (active stage) and the reduced tonus of the smooth muscles of the blood vessels (released endothelin and neuropeptide Y). Knowledge of the physiological mechanisms of erection made clinical diagnosis of their disorders and successful treatment of some forms of impotence possible. PMID- 9333508 TI - [Health risks and economic costs associated with obesity requiring a comprehensive weight reduction program]. AB - An increasing prevalence of obesity all over the world reflects a lack of effective measures in both prevention and treatment of obesity. Obesity as a disease has been underestimated by the lay-public as well as health care providers. However, obesity represents a substantial health problem associated with a decreased quality of life. Obesity is linked to numerous chronic diseases (cardiovascular diseases, diabetes, hyperlipidemia, gout, osteoarthritis, gall stones, and bowel, breast and genitourinary cancers) that lead to premature disability and mortality. Health risks increase with a body mass index (BMI) over 25 in individuals 19-35 years of age and with a BMI over 27 in those 35 years of age and older. Health risks also increase with an excess accumulation of visceral fat manifested as an increase in waist circumference (> 100 cm) or in waist to hip ratio (> 0.85 for females and > 1.00 for males). According to studies carried out in different countries current economic costs of obesity represent 5-8% of all direct health costs. In contrast, effective treatment of obesity results in a substantial decrease in expenditures associated with pharmacotherapy of hypertension, diabetes, hyperlipidemia and osteoarthritis. Both scientists and clinicians involved in obesity research and treatment recommend to introduce the long-term weight management programs focussing more on the overall health of the participants than the weight loss per se. Therefore, it will be necessary to establish new realistic goals in the obesity management that reflect reasonable weights and recently experienced beneficial health effects of modest (5-10%) weight loss. Comprehensive obesity treatment consisting of low fat diet, exercise, behavioral modification, drug therapy and surgical procedures requires differentiated weight management programs modified according to the degree and type of obesity as well as to current health complications present. The Czech Society for the Study of Obesity defined a comprehensive weight reduction program carried out in weight reduction clubs, out-patient obesity clinics and in specialized departments attached to the university hospitals. In order to provide an integrated knowledge from many different disciplines connected with obesity three week postgraduate course has been organized for physicians involved in obesity management. Even the most spread weight reduction clubs in our country (STOB) are supervised by the trained counselors. The main goal of different weight management programs is to find out optimal approaches leading to long-term beneficial outcome and ameliorating the variety of disorders associated with obesity. PMID- 9333509 TI - [Decrease in cardiovascular disease mortality in the Czech Republic from 1984 to 1993 and its possible causes]. AB - BACKGROUND: The objective of the investigation was to evaluate the ten-year development of the cardiovascular mortality rate in two population groups in the age bracket from 25 to 64 years, i.e. in subjects living in six districts which participated in the international WHO project MONICA and in the population of the whole Czech Republic. METHODS AND RESULTS: Data on the mortality rate in 1984 1993 for the age group from 25-64 years were provided by the Institute of Health Information and Statistics, information on the prevalence of risk factors was obtained in three cross-sectional studies implemented in six districts as part of the MONICA project in 1985, 1988 and 1992. In the mortality rate per 100,000 population in the six districts the following changes were revealed (in parentheses the values for 1984 and 1993 are given): men - a statistically significant declining trend in the from all caused mortality (849.3-742.5; p < 0.001) and cardiovascular mortality (367.2-280.4; p < 0.001) and cerebrovascular mortality (69.7-44.8; p < 0.001). In the mortality from ischaemic heart disease (215.7-170.6; ns) a declining trend was not recorded. In women aged 25-64 years in the six districts there was a statistically significant decline of the mortality from all caused (359.5-322.1; p < 0.001), the cardiovascular mortality (115.6-100.6; p < 0.001) and cerebrovascular mortality (31.1-23.6; p < 0.001). The mortality from ischaemic heart disease did not change (49.2-48.8; ns). In the population of the Czech Republic in men the following were detected: a drop of the from all caused mortality (907.1-784.8; P < 0.001), the cardiovascular mortality (383.5-308.4; p < 0.001) and cerebrovascular mortality (76.5-55.3; p < 0.001). Also in women of the Czech Republic a decline of the mortality from all caused was recorded (390.1-328.5; p < 0.001), the cardiovascular mortality (135.3 103.8; p < 0.001), ischaemic heart disease (58.0-48.6; p < 0.001) and cerebrovascular mortality (43.5-27.4; p < 0.001). In 1990 an increased cardiovascular mortality was recorded in men different from the trend during 1984 1993, statistically significant in the Czech Republic (p < 0.05) and in the six districts (p < 0.05). The reasons of this trend are not clear. The role of health services in the mortality drop is not clear, although available data indicate their improvement. Favourable changes were found in risk factors: during the period from 1985-1992 the prevalence of hypercholesterolaemia declined significantly in men and women, the prevalence of hypertension in women and the prevalence of smoking in men declined in the six districts. From nationwide data ensues that after 1989 significant changes occurred in the diet of the Czech population. The meat consumption declined by 1993 by 13%, the milk and dairy product consumption by 26.8% the butter consumption by 43.6% the consumption of vegetable fats increased by 16%, of vegetables by 8%, tropical fruit by 43.2%. These changes probably had an impact on the cholesterol level and BMI of the Czech population. CONCLUSIONS: In the declining cardiovascular mortality trend during 1984-1993 the following may have participated: improved medical care, dietary changes, improvement of the risk profile and other, in particular socioeconomic factors. With regard to the close temporal association of the investigated changes it may be assumed that this development is at least partly associated with changes of the political and economic position in the Czech Republic after 1989. PMID- 9333510 TI - [Cholesterol and triglyceride levels and their development from 2 to 17 years of age]. AB - BACKGROUND: High blood cholesterol levels is important risk factor of an early atherosclerosis and ischemic heart disease. Information on cholesterol level in children in essential for grading the risk and following its trends in the Czech population and for starting an effective prevention already in childhood. METHODS AND RESULTS: Fasting venous cholesterol and triglyceride levels were measured in 1378 children and adolescents, 707 boys and 671 girls aged 1-17 years. Children with familiar hypercholesterolemia or with another illness influencing the measured data were excluded. The mean total cholesterol (TC) level was 4.46 +/- 0.92 mmol/l. It increased from the youngest age, culminated at the age of 13 and decreased afterwards. Mean HDL cholesterol value was 1.23 mmol/l. The lowest value was measured in the youngest age group. The highest values were reached at the age of 11 years and later. TC/HDL ratio was higher than 4.0 only between the age of 3 and 5. LDL cholesterol mean value was 2.70 +/- 0.92 mmol/l. The lowest values were found between 1 and 3 and 13 to 17 years. Triglyceride mean value was 1.22 +/- 0.79 mmol/l. It ranged from 1.53 mmol/l in the youngest age group to 1.03 mmol/l at the age of 7 to 9 years. The values in boys did not differ from those in girls. TC was higher in children with an increased body mass than in children with normal body weight. We did not prove any regional difference in the measured values. CONCLUSIONS: The desired TC value (< 4.41 mmol/l) was found in only 52% of children. In total 18% of all probands, but 26% of school age children belong into high risk category with TC above 5.20 mmol/l. PMID- 9333511 TI - [The importance of iron in humans]. AB - The authors submitted a very brief review of some biological properties of iron in particular protein-bound iron and on the immense amount of iron which surrounds us (it is the fourth most frequent element). In the human organism there is no physiological system for iron elimination. Therefore its absorption is regulated and restricted. Some substances promote its absorption, other inhibit it. An important factor in the regulation of iron absorption are the iron reserves of the organism, the amount of dietary iron and the enhanced erythropoiesis (incl. the ineffective one), while reduced erythropoiesis does not affect iron absorption. Ascorbic acid forms chelates with iron and thus remains soluble and can be absorbed despite the alkaline pH in the duodenum. Unbound trivalent iron remains insoluble. An important asset to rational iron therapy are Lauberger's discovery of ferritin (1936), assessment of the iron plasma level (Heilmeyer and Plotner, 1937), Laurell's discovery of transferrin (1947), and Addison's RIA method for assessment of the serum ferritin level (1974). Sideropenia remains the most widespread deficiency in human pathology, sideropenic anaemia the most readily diagnosed anaemia which is, however, most wrongly treated. PMID- 9333512 TI - [Dialysis therapy in the Czech Republic in 1995]. AB - 1. The number of haemodialyzation centres has more than doubled in the course of five years which increased the dialyzation capacity and at present no patient in the Czech Republic should be denied dialyzation treatment on technical or financial grounds. 2. The number of newly enlisted patients "from the street" remains unfavourable, one third, who are not prepared for this financially pretentious treatment and who have many complications. General practitioners and diabetologists should refer in time patients with chronic renal failure (with a creatinine level above 300 mumol/l) to nephrological--predialyzation clinics. 3. The dialyzed population is aging and in the dialyzation programme polymorbid patients, specially diabetics are increasing in numbers. Therefore the mortality has a rising trend and this will persist. The most frequent cause of death are, similarly as in other countries, cardiovascular complications. 4. The development of another method for the treatment of renal failure is promising--peritoneal dialysis which is used in treatment of almost 5% patients. 5. The number of patients treated by HD per 1 million population is relatively high but only because other therapeutic methods applied in renal failure are not used in a sufficient number of patients. This results in a lower rate of patients treated by RRT methods (HD+PD+TPL)/1 million population, as compared with advanced European countries. PMID- 9333513 TI - [Tics and Tourette's syndrome]. AB - This article is reviewing current knowledge of Tourette's syndrome (TS) including its clinical picture, pathophysiology, genetics and treatment. The semiology of TS is characterized with various types of motor and vocal tics and associated pathological behaviors as attention deficit hyperactivity disorder and obsessive compulsive disorder. Recent pathophysiological and genetic data confirm organic character of the condition. Treatment strategies are proposed with regard to prevailing type of symptoms. PMID- 9333514 TI - [Controlled clinical study of Consupren versus cyclophosphamide in chronic glomerulonephritis. II. Adverse effects]. AB - BACKGROUND: The second part of the study was designed to assess Consupren side effects. METHODS AND RESULTS: The groups of patients studied were described in Part I. Side affects typical of Cy-A were evaluated only in the CS group. Gastrointestinal intolerance, only mild and temporary, was observed in 31%, neurotoxicity in 44%, hypertrichosis in 37%, nephrotoxicity in 25%, and gingival hypertrophy in 19%. Mean values of systolic and diastolic blood pressure did not change significantly in the course of treatment. When changes in blood pressure were individually investigated in particular patients, they were found in 31% in the CS group and in none in the K group. Mean values of uric acid non significantly increased in the CS group and, on individual investigation, hyperuricaemia was observed in 31%. Mean values of serum potassium did not alter significantly. Signs of possible hepatotoxicity were found in 37% patients of the CS group. In this group, there was a significant decrease in haemoglobin mean values and a decrease in haemoglobin of more than 25 g/l was observed in 44% of CS group patients. In the K group significant decrease in mean leukocyte count was noted, but no patient developed real leukopenia. CONCLUSIONS: The occurrence of side effects was comparable to data known from the literature. PMID- 9333515 TI - [The effect of dialysis solutions containing amino acids on the nutritional status of patients treated with continuous ambulatory peritoneal dialysis]. AB - BACKGROUND: In the patient population which is subjected to haemodialysis or peritoneal dialysis is a high prevalence of protein-energy malnutrition which independently on the causal factor causes a deterioration of their prognosis. To influence protein nutrition in patients on peritoneal dialysis Nutrineal was developed, i.e., a dialysis solution using as an osmotic agent a 1.1% amino acid mixture instead of glucose used as a rule. The objective of the present study was to evaluate the effect of its administration on some nutritional indicators in patients treated by continuous ambulatory peritoneal dialysis (CAPD) and to assess its tolerance. METHODS AND RESULTS: The authors investigated eight patients aged 45.5 years (29-78 years, median, minimal and maximal value), treated for 19.9 (5-42) months by CAPD. For a period of four weeks Nutrineal was administered once a day instead of glucose based solution to patients without diseases complicating nutrition, with a serum albumin concentration (ALB) below 35 (20-34) g/l. Before treatment and after its termination an anthropometric examination was made, the rate of protein catabolism was examined (PCR), plasma concentrations of total proteins were assessed, as well as ALB, transferrin (TRF) and 14 free amino acids substituted by the dialysis solution. For statistical comparison the paired Wilcoxon test was used. As compared with the baseline value of 0.83 g/kg/24 h after treatment a significant increase of PCR was recorded--to 0.96 g/kg/24 h (p < 0.05) as well as a significant increase of the urea concentration from 18.4 (10.9-34.8) mmol/l to 26.7 (19.6-33) mmol/l (p < 0.05). The value of phosphorus declined significantly from 2.25 (1.6-2.6) to 1.9 (0.9 2.5) mmol/l (p < 0.05). No significant difference was recorded in the anthropometric findings and in concentrations of total proteins, ALB, TRF and amino acids. CONCLUSIONS: Four weeks administration of a solution with amino acids raised significantly the PCR which may be indirect evidence of an anabolic effect and it reduced plasma phosphates which participate in uraemic toxicity. The solution was well tolerated by the patients. Final evaluation of the values of the solution containing amino acids calls for long-term and controlled studies. PMID- 9333516 TI - [Maturation and fertility in women with low birthweights]. AB - BACKGROUND: Some 6% infants are born with low birthweights (less than 2500 g). Due to better obstetric and neonatalogical care the percentage of surviving infants is increasing. Sexual maturation of girls, menarche, the course of their gestation and fertility are not frequently investigated. That is the objective of the present work. METHODS AND RESULTS: The author evaluates two groups of women with birthweights lower than 2500 g: 1. 180 women born at the Gynaecological and Obstetric Clinic in 1995-1957. 2. 435 women dispensarized in the welfare centre for child and adolescent gynaecology at the same Clinic. Women of the control group born in 1969-1972 could not be evaluated because the addresses were not available. CONCLUSIONS: The examined women with a lower birthweight reach an average height in adult age and the mean body weight is in the lower half of normal values for our population. Sexual maturation is harmonious, only slightly retarded, menarche is by 3-6 months delayed. The course of pregnancy and delivery is as a rule within the normal range, however the number of infants born with a birthweight of less than 2500 g is double. Therefore greater care during pregnancy is indicated. PMID- 9333518 TI - Insurers keep genetic test options open. PMID- 9333517 TI - [Reference values, metabolism and findings of abnormal values of magnesium in childhood]. AB - In the introduction we summarize specifics of reference values and metabolism of magnesium in childhood. We studied the frequency of abnormal values of magnesium in both children and adults hospitalized in Faculty Hospital Motol, Prague. We analyzed 6761 tests of S-Mg and 356 dU-Mg results located in archives of Department of clinical biochemistry (1. 1. 1992-1. 11. 1995). The frequency of low level of S-Mg (< 0.65 mmol/l) was 14%. Hypermagnesemia was present in 1.6% of the total number of cases. We found low values of dU-Mg in 20% of the total number of cases, hypermagnesiuria was present in 8.4% cases. We also proved that hypomagnesemia is more frequent in both boys and men than in girls and women. Critical values of S-Mg below 0.5 mmol/l are less frequent (84 cases). Estimated frequency of clinical significant hypomagnesemia in hospitalized patients was about 1.25%. PMID- 9333519 TI - Europe and FDA agree to limited inspections. PMID- 9333520 TI - Summer recess might squeeze FDA reform bill. PMID- 9333521 TI - "Dr. Peter" lives on through AIDS daycare centre. PMID- 9333522 TI - [Ethical education: experience and ethical reflexion]. PMID- 9333523 TI - Lawsuits over laser patents raise serious issues for physicians, patients. PMID- 9333524 TI - Confronting despair: the Holocaust survivor's struggle with ordinary life and ordinary death. PMID- 9333525 TI - Unvarnished viewpoints and scientific scrutiny. Letter to the editor provide a forum for readers and help make a journal accountable to the medical community. PMID- 9333526 TI - Issue dedicated to Dr. G. T. Young on the occasion of his 80th birthday. PMID- 9333527 TI - Proceedings of the 45th annual meeting of the Congress of Neurological Surgeons, honoring John A. Jane, M.D., Ph.D. San Francisco, California, October 14-19, 1995. PMID- 9333528 TI - Biology of the Synovial Joint. Cardiff, United Kingdom, 30 June-2 July 1996. Abstracts. PMID- 9333529 TI - [Normal values for 24-hour manometry of the esophagus]. AB - OBJECTIVE: To obtain normal values of 24-hour manometry of the oesophagus. SUBJECTS AND METHODS: Oesophageal pressures were measured in 41 healthy volunteers who had given informed consent. Recordings were made for 24 hours via a two-channel catheter in 27 and via a 4-channel one in 14 subjects. The catheter orifices were 5 and 15 cm respectively 5, 10, 15 and 20 cm above the lower oesophageal sphincter. RESULTS: Median of contractions was 1523 at 5 cm and 1500 at 15 cm (1635 at 10 cm and 2135 at 20 cm) contraction amplitudes were 31 mm Hg at 5 cm, 26 mm Hg at 15 cm; 26 mm Hg at 10 cm and 37 mm Hg at 20 cm. On average 44% of the contractions were propulsive, 17% simultaneous and 30% nonpropulsive, the remainder not clearly defined. Neither age nor sex had a significant influence on the results. Motor activity was reduced during sleep. During eating the number of contractions, their amplitude and propulsive force increased. CONCLUSION: The listed measurements, by defining normal values, make it possible to diagnose hypo- and hypermotility of the oesophagus during long-time manometry. Two-point measurement is sufficient for assessing the smooth-muscle component. PMID- 9333531 TI - [Diagnosis and therapy of thyroid carcinoma]. PMID- 9333530 TI - [Space occupying lesion in the anterior mediastinum in a patient with Basedow disease and previously diagnosed osteosarcoma]. AB - HISTORY AND CLINICAL FINDINGS: A 24-year-old woman with osteosarcoma of the right thigh, diagnosed 12 years ago, complained at a follow-up examination of decreased exercise tolerance, increased nervous tension, heat intolerance, weight loss, hair loss and irregular stools. Examination revealed tachycardia (100/min), mild exophthalmus and a small goitre. INVESTIGATIONS: A decreased basal TSH level (0.002 mU/ml), raised peripheral thyroid hormone (fT4 6.7 ng/dl, total T3 7.8 micrograms/l) and a TSH receptor antibody titre of 33.4 U/l) were compatible with immune type Graves' disease. Radiology revealed an upper mediastinal space occupying lesion which scintigraphically was separate from thyroid tissue. A metastasis of the osteosarcoma or thymus hyperplasia were considered the most likely cause. TREATMENT AND COURSE: The mediastinal lesion regressed under thyrostatic treatment with carbimazole (20 mg daily by mouth). But the clinical picture, localization and negative scintigraphy provided the diagnosis of transitory thymus hyperplasia in the course of Graves' disease. CONCLUSION: In immune type Graves' disease with a mediastinal space-occupying lesion, not only intrathoracic goitre but also thymus hyperplasia should be considered in the differential diagnosis. PMID- 9333532 TI - [Reperfusion therapy in acute myocardial infarct, Thrombolysis or balloon dilatation?]. PMID- 9333533 TI - [Arteriosclerosis--a Chlamydia pneumoniae infection]. PMID- 9333534 TI - [Chlamydia pneumoniae and atherosclerosis]. PMID- 9333535 TI - [Impingement syndrome]. PMID- 9333536 TI - [Diphtheria re-vaccination of adults]. PMID- 9333537 TI - [Course of blood levels of calcium, inorganic phosphate, alkaline phosphatase, parathyroid hormone and calcidiol (25-OH-D3) in one and two year old thoroughbred horses]. AB - The present study was aimed to determine the contents of calcium, inorganic phosphate, parathormon, 25-OH-D3 and the activity of alkaline phosphatase in the plasma of one- and two-years-old thoroughbred horses. Data were obtained monthly from 44 one-year-old thoroughbred of 4 different studs from May during grazing season and from October during stable-, resp. training-season up to april of the following year. Calcium, inorganic phosphate and the activity of alkaline phosphatase were measured with a photometric method and the concentration of PTH and 25-OH-D3 were determined with a radioimmunoassay. The following results were obtained: Calcium: The concentration of calcium in the plasma of one-year-old thoroughbred horses was 3.03 +/- 0.23 mmol/l during grazing-season and 3.14 +/- 0.14 mmol/l during the following stable-, resp. training-season. Inorganic phosphate: The concentration of inorganic phosphate was significantly affected by the age. The average was 1.7 +/- 0.19 mmol/l during grazing-season and 1.3 +/- 0.19 mmol/l during the following stable- and training-season. Activity of alkaline phosphatase: The activity of alkaline phosphatase was also significantly affected by the age. The average of the activity was 403 +/- 86 U/l during grazing-season and 308 +/- 65 U/l during the following winter period. Parathormon: There were big differences between the averages of parathormon during grazing-season (1.27 +/- 0.45 ng/ml) and the following winter-season (0.9 +/- 39 ng/ml). Besides from that there were big individual differences. 25-OH-D3: The concentration of 25-OH-D3 during grazing-season (10.38 +/- 3.08 ng/ml) was lower than during the winter period (13.03 +/- 2.86 ng/ml). The significance of the obtained results is discussed in relation to the corresponding literature. PMID- 9333538 TI - [Natural and synthetic glucocorticoids in the racing horse: a review of the literature]. AB - This review compromises data about endogenous cortisol and its physiological variations in horses. The influence of synthetic glucocorticoids on the endogenous cortisol concentrations is discussed as well. The second part of the review summarizes detection times of therapeutically used glucocorticoids (dexamethasone, betamethasone, triamcinolone, prednisone, prednisolone, methylprednisolone and hydrocortisone) in the horse and their implication for doping control. PMID- 9333539 TI - [Histological and histochemical investigations on the structure of udder skin of cattle with special reference to changes during in vivo udder perfusion models]. AB - Generally, the skin of the bovine mammary gland is found to be similar to the skin of the integument. In the epidermis, the stratum lucidum is lacking. A prominent stratum corneum can be observed as well as in the haired areas and in areas without hairs as well. Distinct granules of melanin are located mainly in the stratum basale in pigmented skin areas. By means of the alkaline phosphatase, dendritc cells can be seen in the basal parts of the epidermis. In the corium, collagenous fibres are less thick below the epithelium, whereas coarse bundles are seen deeper in the stratum reticulare. The appearance of multilobal sebaceous glands and apocrine sweat glands is associated with a hair follicle, which occur loosely distributed as single hairs. A large erector pili muscle is situated on the side of the hair slope. The numerous sweat glands with wide diameters appear below the hair papilla, and vary greatly in shape and size. Myoepithelials cells are seen at the periphery of its secretory portions. The small number of differences in all areas of the skin examined, are reduced to differences between haired areas and the base of the teats. The histologic structure of the epidermis remains unchanged by the perfusion. A partly collagenolysis occur in the dermis and subcutis. PMID- 9333540 TI - [Crystalluria in sows]. AB - In 12 breeding sows the influence of high feed levels of calcium (Ca 16.0 mg/kg, P 5.9 mg/kg, Ca/P ratio 2.71:1), of a mineral feed mixture (Ca 13.8 mg/kg, P 8.3 mg/kg, Ca/P ratio 1,66:1) and of phosphorus (Ca 7.0 mg/kg, P 11.0 mg/kg, Ca/P ratio 0.64:1) on blood concentrations and renal excretion of minerals (Ca, P, Mg), electrolytes (Na, K) as well as development of urine concrements (crystalluria) was investigated in comparison to a control feed (Ca 7.3 mg/kg, P 6.0 mg/kg, Ca/P ratio 1.23:1). Besides the effect of water supply on formation of crystalluria was tested. Studies showed that especially high levels of phosphorus in the feed are responsible for excretion of urinary crystals. Sediment consisted of Ca phosphates mainly, which could be detected as amorphous crystals microscopically. Alkaline pH values in urine and an insufficient water supply supported development of crystalluria, but formation of crystals differed greatly between individuals. Cystoscopic investigations demonstrated inflammatory alterations of the bladder mucosa in sows with crystalluria. Therefore crystalluria in sows has to be considered as a risk factor for urinary tract infections. PMID- 9333541 TI - [Investigations of raw milk for Shiga-like toxin producing Escherichia coli by the polymerase chain reaction]. AB - This is a report about the occurrence of Shiga-like toxin (SLT) producing Escherichia coli in milk in southern Sachsen-Anhalt. Samples were taken from tanks of milk at the farms. The samples were investigated by polymerase chain reaction as recommended by the "Bundesinstitut fur gesundheitlichen Verbraucherschutz und Veterinarmedizin". Using PCR, in 11% of the tank samples sequences coding for SLT I and/or SLT II were detected. 75% of the as positive detected samples had eae gen. Some cows in these farms spread potentially enterohemorrhagic E. coli. These farms should not sell milk for raw consumption. PMID- 9333542 TI - [Importance of blood serological examinations in the context of rhinitis atrophicans diagnosis]. AB - In a period of three years in 95 breeding-herds, which were free from Rhinitis atrophicans (R. a.), 5001 blood-samples were examined. All samples were examined by the SNT/EBL-cell-culture-test-mostly however by the SNT/ELISA-system- and were free of antibodies against R. a. On the contrary in 6 herds, that had bought swines from latent infected populations, antibodies against the toxin could be found before clinical signs were to be seen and before toxin producing pasteurellas could be discovered, too. In other 4 herds antibodies against R. a. could be found. In the last mentioned herds R. a. was suspected by clinical, bacteriological and pathomorphological examinations. The serological determination of blood may replace the provement of the diameter of the nose by pathomorphological diagnosis. The serological investigation is a good test. After positive results of antibodies comprehensive bacteriological tests should follow. This can be integrated in the diagnostical system of R. a. very easily, because enough blood samples will be taken ordered by the official examinations for European Swinefever- and Aujeszky-disease. PMID- 9333543 TI - [Examination of antigen preparations from the virus of enzootic bovine leukosis with regard to suitability for immunoblotting]. AB - 8 different leukosis antigen preparations for immunodiffusion from 6 European countries were examined regarding their suitability for immunoblotting. Emphasis was made on demonstration of immunoreactions at 51 kDa beside reactions at 24 kDa. The whole protein content of the different preparations varied very much. gp 51 could be detected only by monoclonal antibodies at a molecular weight of 53 kDa, in one preparation at 26 kDa. With sera from field or with the European reference serum E4 none or only weak reactions could be observed at these molecular weights. All antigen preparations contained bovine IgG that is divided in L- and H-chains by SDS-electrophoresis under reducing conditions. These single chains reacted also with anti-bovine-IgG-biotin conjugate und disturbed the detection of the leukosis specific reactions. The bovine IgG could be removed by a Protein-G-column. One of all tested leukosis antigens, which are prepared for immunodiffusion, was suitable for immunoblotting. This antigen preparation contained only few bovine IgG and had strong reactions at 24 kDa. A special antigen preparation for detection of other reliable immunoreactions in the immunoblot has to be developed. PMID- 9333544 TI - [Animal nutrition in veterinary medicine--actual case reports. Nutritionally related disturbences in bone development in emus and rheas]. AB - Two cases of disturbed health and growth of the skeleton in emus and rheas caused by faults in feeding and mineral supply are reported. 13 of 37 emus developed (beginning in the 3rd week) more and more signs of perosis. The diet fed in this case was based mainly on pelleted diets for piglets and rabbits and unpelleted supplements. The ingesting behaviour (selective intake and refuse of fines and supplements) resulted in an imbalanced mineral intake. In the second case 3 of 15 young rheas showed unphysiological postures of the necks vertebral column. Disposed by an absolutely insufficient calcium supply (the real ingested diet was based on corn and white bread mainly) fractures of vertebrae were provoked by unprofessional handling. During the healing process (uncalcified) connective tissue near to fractures localization led to the abnormal posture of the neck. PMID- 9333545 TI - [Chronic hallucinatory psychosis and late onset schizophrenia: the same entity?]. AB - The distinction between schizophrenia and chronic delusion syndromes (such as paraphrenia, late-paraphrenia and "Psychose Hallucinatoire Chronique") is currently used in France, although there is no international criteria (ICD 10 or DSM IV) for chronic delusion syndromes. It is thus worth analysing the literature in order to compare the differences between late-onset schizophrenia and "Psychose Hallucinatoire Chronique", and the similarities between young-onset schizophrenia and chronic delusion syndromes. Clinical investigations clearly differentiate "Psychose Hallucinatoire Chronique" and late-onset-schizophrenia from young-onset schizophrenia because they have more delusion and hallucinatory symptoms, less negative symptoms, better evolution, and better sensitivity to antipsychotic drugs. Epidemiological data show that "Psychose Hallucinatoire Chronique" and late-onset schizophrenia have both a different sex-ratio (around 7 women for 1 man) than young-onset schizophrenia (nearly 1 woman for 1 man), and that in "Psychose Hallucinatoire Chronique" and in late-onset schizophrenia, social withdrawal is frequently observed before onset of the disorder. Lastly, putative risk factors may be shared by "Psychose Hallucinatoire Chronique" and late-onset schizophrenia, and may isolate them from young-onset schizophrenia, for example regarding the oestradiol hypothesis (oestradiol enhance dopamine efficacy and delay the onset of delusion disorders), the impact of sensory handicaps (which may be clinically and experimentally associated with hallucinations), or the role of genetic and familial factors (with a familial concentration intermediate between the familial concentration of schizophrenia of schizophrenic proband, and the familial concentration of schizophrenia of control probands). In accordance with this review of the literature, the authors conclude that the absence of specific criteria for late-onset schizophrenia and/or "Psychose Hallucinatoire Chronique" In international diagnostic manual risk to counter the facility to detect specific risk factors involved in the pathogenesis of schizophrenia. PMID- 9333547 TI - [Comparative study of normal subjects and obsessive compulsive subjects on intrusive thoughts and memory]. AB - Four surveys have shown that more than 80% of normal subjects have obsessive thoughts with content similar to those found in obsessive compulsive patients. "Abnormal" and "normal" intrusive thoughts do not differ in content but in duration, frequency and rejectability. Thirteen DSM III-R non-depressed obsessive compulsive patients with checking rituals were compared with 13 sex-, age-, and education matched control subjects. Our study compared the content of abnormal and normal obsessions. No between-group difference was found suggesting that a sub-group of obsessive compulsive patients with checking rituals did not change the general result. The memory processes were also compared. Several investigators have shown an impairment in visual spatial memory tasks but none in verbal tasks with obsessive compulsive patients. In three studies non clinical checkers were found to show some deficits in memory compared to non checkers. In our study we used the Wechsler memory scale. Clinical checkers appear to have difficulties recalling details of meaningfully linked verbal sequence and immediate visual reproduction. PMID- 9333546 TI - [Capacity to experience pleasure and displeasure by Cloninger and von Knorring in type I and II alcoholism]. AB - The aim of the present study is to test the hypothesis that Cloninger and von Knorring's type II alcoholics are more hedonic and experience less displeasure than type I alcoholics. 55 inpatients filled out the DSM III-R criteria for abuse or alcoholic dependence. The alcoholics were dichotomized into type I or type II using the onset of their alcoholism (type II < 25 years, type I > or = 25 years). In the few days of their hospitalization, the subjects filled out the Fawcett Clark Pleasure Capacity Scale-Physical Pleasure (FCPCS-PP), the Hardy Displeasure Capacity Scale-Physical Displeasure (HDCS-PD) and the abridged form of the Beck Depression inventory (BDI). The sociodemographic characteristics, the proportions of "abusers" and "dependents" and the scales scores were compared between the two groups. The sex-ratio and the educative level were not different between the two groups; the values of the chi 2 were respectively 1.36 (df = 1), p = 0.24 and 1.03 (df = 2), p = 0.59. The ratio "abusers" /"dependents" was not different (chi 2 = 1.115, p = 0.291). The mean age of the type I alcoholics was higher (m = 46.7, sd = 11.2) than that of the type II (m = 38.9, sd = 7.9) (U = 218.5, p = 0.013). Using covariance analyses (ANCOVA) to control the age, the results have shown that the FCPCS-PP and HDCS-PD scores of the type II alcoholics were not different than these of the type I alcoholics [respectively F (1.52) = 1.5, p = 0.23 and F (1.52) = 0.14, p = 0.7]. In conclusion, we failed to confirm the hypothesis that type II alcoholics would experience more pleasure and less displeasure than type I alcoholics. These negative results were discussed and the potential factors that could explain them are detailed. PMID- 9333548 TI - [Sleep disorders in prison]. AB - Prisoners often ask the psychiatrist soporific. After a clinical review, the author exposes the particular difficulty concerning psychotropic distribution patterns in this unusual institution. He puts in an ethical consideration about the concept of "hypnotic deliberately retarded effect". PMID- 9333549 TI - [Development and validation of a perceived stress questionnaire recommended as a follow-up indicator in occupational medicine]. AB - This work was originated from the concern for the availability of a short, acceptable and reliable instrument for self-assessment of perceived stress. The questionnaire was designed to be routinely applied within a visit at an Occupational Medical Center. A new questionnaire was developed with this aim, consisting of 9 subscales presented in a Lickert form, with 4 modes of answer, scored from 0 to 3. The content of the 9 items covers the multiple facets of perceived stress and its consequences: "feeling of being under pressure", "impatience", "irritability", "intrusive thoughts about work", "inability to entertain", "discouragement", "morning fatigue", "food compensation", "compensation by smoking". Stress global score is defined as the sum of the 9 elementary scores. The first 7 items are similar in their construct to the 7 factor solutions of the principal component factor analysis performed on Levenstein Perceived Stress Questionnaire (1993). On the other hand, in comparison with Cohen Perceived Stress Scale (1983), our instrument keeps an important place for affective, physiologic and behavioral impact of stressing situations. A study on the homogeneity of the scale, its factorial structure, and its time reproducibility after 6 weeks of interval, was carried out on a first population of 91 subjects seen during an occupational medical visit in several companies of Paris district (PCV-Metra group). The coefficient of internal consistency is very high (Cronbach's alpha = 0.82). Principal component analysis extracted two factors, which were unchanged after a Varimax rotation and respectively represented 42% and 13% of the total variance: they can be interpretated as a general perceived stress component (being overwhelmed, loss of control) and a behavioral bipolar component opposing food compensation to smoking, whilst facing stressing situations. Test-retest correlation coefficient is 0.88 (Pearson r as well as intraclass correlation coefficient), without any significant gap between the first and the second assessment. A second study was carried out on 761 working individuals seen in the same conditions during an occupational medical visit in the same companies (596 males and 65 females, aged 40.1 +/- 8.8 years). Socio-demographic data analysis showed higher stress scores in females (10.2 +/- 4.0), than in males (8.5 +/- 3.7) (p < 0.0001). Detailed analysis showed differences related to gender in the same direction for the items "irritability", "discouragement", "morning fatigue" and "food compensation". Highest stress scores, for both males and females, were found for the items "intrusive thoughts about work" and "morning fatigue". Stress global score was correlated with socioprofessional status (SPS): unskilled and skilled workers (n = 39) as well as technicians (n = 346) exhibited lower scores than clerical workers (n = 108) or engineers (n = 162) (ANOVA, p < 0.0001). The comparison between mean scores performed separately by gender, given the different sex-ratio according to SPS (employees were mostly females) confirmed the association between stress score and SPS only in males, with blue collars and technicians looking less under stress than engineers. Metrological properties of this Perceived Stress Questionnaire incite to perform studies focused on the associations between such a stress index measured in an occupational setting, and several other clinical or biological variables, especially those supposed to constitute classical cardiovascular risk factors. PMID- 9333550 TI - [Scoring and validation of the Clock Face Test in psychometric assessment of elderly subjects]. AB - AIMS: To propose an original evaluation and validation of the Clock Face Test (CFT). METHOD: Outpatients; 163 elderly people, aged over 65 years. Factory analysis, Student's test. RESULTS: The test shows a positive correlation with Folstein's MMSE (r = 0.769) and Signoret's BEC 96 (r = 0.644), a good specificity (75% with MMSE; 79% with BEC 96) and a good sensitivity (87% with MMSE; 74% with BEC 96). Four factors are identified by factory analysis; 74% of the variancy is explained by the first three factors. CONCLUSION: The CFT is a valid tool, easy to conduct. Several applications are possible for diagnosis, psychometric control and therapeutic trials. PMID- 9333551 TI - [The Young Cognitive Schema Questionnaire: translation and preliminary validation]. AB - Young (1990) proposed a clinical scale to assess personality disorders. We translated and used this scale for validation purpose in pathological (n = 113) and control (n = 54) samples. Our results reflected that Young Schema Questionnaire (SQ) has a good discrimination value between patients (Axes I and II of the DSM III-R classification) and controls. Statistical comparisons indicated significant differences between three subgroups-axis I only (n = 53), axes I and II together (n = 60), and controls (n = 54). The SQ score was the highest in the patients with personality disorders, reflecting the sensitivity of the scale to Axis II pathology. Principal component analysis (unrotated factors) showed a first factor, a failure axis (eigenvalue = 7.93), and a second factor, a narcissistic one (eigenvalue = 1.30). These two principal components explained 62% of the variance. PMID- 9333552 TI - [Administration of high dose levothyroxine in treatment of rapid cycling bipolar disorders. Review of the literature and initial therapeutic application apropos of 6 cases]. AB - INTRODUCTION: Rapid cycling among bipolar disorders was characterized in 1974 by Dunner and Fieve by at least 4 episodes per year, lithium resistance, female predominance. The idea of using thyroid hormones is based on frequent coexisting thyroid dysfunction. Thyroid hormones where first used in the forties, and more recently (table I) in open label and blind trials (Stancer et Persad, 1982; Bauer et Whybrow, 1990). OBJECTIVE: Open label study of levothyroxine in rapid cycling bipolar disorder. MATERIAL AND METHODS: Six subjects (4 females and 2 males, mean age 45.5 years at onset of bipolar disorder) meeting DMS III-R criteria for rapid cycling bipolar disorder were consecutively included in an open label study of levothyroxine. Subject characteristics are presented in table II. RESULTS: After almost two years follow-up results appear positive in 67% of cases (2 complete remissions, 2 partial remissions and 2 failures). CONCLUSION: Our cases, with data reported in the literature, support the potential efficacy of levothyroxine for treatment of rapid cycling bipolar disorder patients. We suggest a protocol for the good application of this potential new treatment (table III). PMID- 9333554 TI - [DSM IV criteria for post-partum depression: debatable?]. PMID- 9333555 TI - [Proceedings of the 6th Lilly Symposium of the Central Nervous System. Paris, France, 22-23 November 1996]. PMID- 9333553 TI - [Fluoxetine, akathisia and suicide]. AB - Several cases of suicidal thinking or suicidal behavior during fluoxetine treatment are described in the literature. Akathisia or dysphoric extrapyramidal reactions may explain the emergence of suicidal ideation during fluoxetine treatment. Retrospective and controlled studies found no relationship between fluoxetine and the emergence of suicidal ideation. Some cases reports of akathisia or suicidal ideation with other selective serotonin reuptake inhibitors (SSRIs) suggest that same mechanisms (pharmacokinetic or pharmacodynamic interactions) are involved in extrapyramidal effects during treatment with SSRIs. The clinical use of SRIs is discussed. PMID- 9333556 TI - [Morphologic plasticity as a component of cognition]. AB - Two recent developmental genetics studies suggest an important role for transcription factors of the homeodomain protein class in the morphogenesis of cerebral structures and the form and extent of the various domains that constitute the nervous system (Krumlauf, 1994; Rubenstein and Puelles, 1994). Our team recently took a look at the hypothesis that the same transcription parameters have a function in axonal growth and recognition of paths and targets. It has been shown that cortical or spinal medullary neurons are capable of responding, by a change in axonal growth rate, to regulation of the activity of certain transcription factors (Prochiantz and Theodore, 1995). This demonstration was facilitated by a particular property of the DNA binding region of the transcription factors. This property consists in crossing plasmic membranes and addressing cell nuclei directly (Prochiantz, 1996). The homeodomain proteins are expressed not only during development but also in adulthood. It is possible that their continuous expression or reactivation may be associated with the morphological changes in neurons observed in the mature nervous system. The latter hypothesis will be discussed on the basis of our current knowledge of the expression and role of the transcription factors. The authors will draw some conclusions regarding the possibility of considering these nuclear factors as agents of cerebral plasticity and as targets of potential pharmacological value. PMID- 9333557 TI - [Indications for electroconvulsive therapy]. AB - ECT, in which first experiments were made by the italian Cerletti more than half a century ago, underwent, in the seventies, a definite decline, as it was less and less applied to patients, a result of the influence of anti psychiatry. During the last fifteen years, there has been a legitimate renewal of the interest for this therapy; its indications seem now well codified and its techniques and practises have evolved considerably. Actually, in order to carry out ECT under general anaesthesia, it is necessary to have a pluridisciplinary team, assembling nurses, anaesthesists and psychiatrists that will use more and more effective appliances and adequate anaesthetics. Many of the parameters able to influence ECT's effectiveness are now well known and can be used and adapted according the individual characteristics of each patient. These parameters are: the lateralisation of the electrodes, the intensity of the electric current, the duration of the epileptic fit, the modification that appear in electroencephalography and the frequence of the sessions. According to different investigations, it seems that we must systematically question the medical treatments we associate to ECT. For instance, it is highly recommended not to prescribe with ECT benzodiazepines or antiepileptic mood stabilizers, while antidepressants or neuroleptics do not seem to exert any influence on the effectiveness of the treatment. Some authors think caffeine and triiodothyronin (T3) could have an interesting effect when combined with ECT. As to the indications of shock therapy, they can be now more and more precisely defined making of this treatment an indispensable instrument in the cure of depressive disorders. But ECT is also appropriate in maniac disorders once neuroleptic treatment has failed or else in the very beginning in highly acute cases, and mainly in mixed episodes for which medical treatment is often difficult to adapt. In schizophrenia, ECT can also be prescribed in definite circumstances as catatonia, paranoid states or schizoaffective episodes. Therefore, ECT constitutes a safe and comfortable therapy for the patient since its side effects are essentially characterized by cognitive disorders, and its main contraindications consist of severe cardiovascular diseases. ECT is also an essential tool in some definite cases. PMID- 9333558 TI - [Are convulsions necessary for the antidepressive effect of electroconvulsive therapy: outcome of repeated transcranial magnetic stimulation]. AB - Sismotherapy (ST) brings about numerous neurobiological changes, particularly changes in neuromediators and their receptors, second messengers, neuropeptides and neurotropic factors, a number of which are hypothesized to play a role in the pathophysiology or therapeutics of affective disorders (M. Fink). What is not yet known is which of these mechanisms is crucial for the psychotropic and anticonvulsant effects of ST. However, it is clear that the effects of ST tend to be relatively acute, and do not attack the deep-seated abnormalities that are the underlying causes of recurrences of affective disorders. This is corroborated by the fact that in animals, most of the effects of ECS on catecholamines and their receptors (and on receptors for benzodiazepines or neuropeptides such as TRH) tend to be relatively transient, and in most cases have been found to represent compensatory adaptations to the induced motor convulsions. However, recent preclinical data using attenuation, and clinical findings using reiterated transcranial magnetic stimulation (rTMS), suggest that it may not be necessary to provoke a clonic convulsion in order to achieve the beneficial psychotropic and anticonvulsant effects of ST. In rodents receiving stimulation to the cerebellar tonsil, seven daily subacute low-frequency sessions (stimulation at 1 Hz for 15 minutes) produced clear improvement in clonic convulsions and in post-discharge thresholds, together with durable inhibition of convulsions when stimulation was resumed (Weiss et al., 1995). Stimulation at 1 Hz for 15 minutes was more effective than stimulation at 10 or 20 Hz in attenuating convulsions. Although reiterated ECS also induced an anti-triggering effect, this dissipated rapidly over five days (Post et al., 1984). It is of great interest that recent publications have shown that rTMS at 10 or 20 Hz to the left frontal cortex, administered to patients suffering from refractory depression (George et al., 1995) or to patients (hospitalised or not) with milder degrees of depression (Pasquale-Leon et al., 1996), had a moderate or marked antidepressant effect. In these studies, rTMS showed few unwanted effects (other than mild pain in some patients, due to contraction of the temporal muscles); it did not induce motor convulsions, and did not, as such, appear to be associated with the memory loss described in subjective accounts or in preliminary neuropsychological tests (Little and Kimbrell et al., 1996). The optimal frequencies, durations and positions for rTMS to maximise its antidepressant effect still remain to be determined. However, the first controlled and open studies have tended to show that (because of the capacity of rapid magnetic fluxes to produce sub-convulsant electrical discharges that are relatively localised in the brain), rTMS may be found to be a clinically useful antidepressant model. This would suggest the possibility that some of the neurochemical changes induced by the clonic convulsions of ECS could be directly induced by stimulation at the very edge of the threshold (but still below it); this would open up the hope that one day these endogenous neurochemical processes could be identified and exploited in an optimal way for therapeutic purposes. PMID- 9333559 TI - [Anger outbursts in unipolar depressive disorders]. AB - Approximately one third of depressed outpatients present with "anger attacks", sudden spells of anger accompanied by symptoms of autonomic activation such as tachycardia, sweating, flushing, and tightness of the chest. These anger attacks are experienced by the patients as uncharacteristic of them and inappropriate to the situations in which they occur. Depressed patients with anger attacks are significantly more anxious and hostile, and they are more likely to meet criteria for borderline, histrionic, narcissistic, and antisocial personality disorders than depressed patients without anger attacks. Treatment studies suggest that antidepressant treatment of anger attacks in depression is helpful and sage. Anger attacks disappear in 53-71% of depressed outpatients treated with antidepressants such as fluoxetine (Prozac), sertraline and imipramine. In addition, the rate of emergence of anger attacks after treatment with fluoxetine (Prozac) (6-7%) is no different from the rates observed after treatment with sertraline (8%) and imipramine (10%), and lower than the rate with placebo (20%). Finally, one can hypothesize that antidepressants that affect serotonergic neurotransmission, known to be involved in the modulation of aggressive behavior in animals and humans, should be particularly effective in this population. Larger placebo-controlled studies, comparing selective serotonin reuptake inhibitors such as fluoxetine with relatively noradrenergic tricyclic antidepressants such as desipramine, may help us understand whether depressed patients with anger attacks show a distinctive responsiveness to drug treatment. PMID- 9333560 TI - [Anxiety, aggression, agitation and depression: psychopathologic aspects]. AB - While clinical experience has long since shown that there are different types of depression, in particular anxious and hostile depressions, the psychopathological analysis of the various forms remains of current interest. At least four psychopathological models are currently available. The first raises the question of the continuity between reactions to separation, particularly studied in children, and the clinical expression of certain forms of depression in adults. In many aspects, the latter suggest the sequence: protest-despair-detachment. The second model raises the question of the relationship between the depressive disorder and the organization of certain personalities. The considerable comorbidity between the borderline personality and affective disorders suggest that these two different disorders share a common dimension. The third-cultural model hypothesizes a relationship between the sociocultural prohibition of aggressive responses and the incidence of depression. The fourth model is based on the existence of a specific biological constraint related to abnormalities of serotonin metabolism, to which dysregulation of anxious and aggressive-impulsive behavior patterns during depression are considered related. In conclusion, it may be considered that several models, in particular that of reaction to separation, may, at least in part, account for "positive-expression" depression, but that at least two questions have still to be answered: that regarding the relationships between depression and personality, and that concerning the relationships between the psychopathological constraints related to serotonin metabolism dysfunction and the "positive" expressions. PMID- 9333561 TI - [MRI and cognition]. AB - Many groups are trying out functional MRI (fMRI) to study normal and diseased brain function. FMRI is a technique that images intrinsic blood signal changes accompanying variations of cerebral blood flow (CBF), cerebral blood volume (CBV) and oxygenation during mental activity. The main advantages which account for fMRI's increasing popularity are its time resolution (less than 1 second), a fine spatial resolution (on the scale of 1 mm), and the ability to acquire, within the same individual, repeated, multiple scans in a non-invasive manner. While the basic design of most studies centers on comparison of baseline to an active (test) condition, as the Positron Emission Tomography (PET), fMRI allows an important departure from the single scan per condition study-design, into more flexible paradigms of multiple scans over varying time intervals and multiple conditions. Although the technique is only a few years old, a large volume of experimental data have already been published. In this presentation we would like to provide potential users with a short overview of the basic principles and the main applications in cognitive neuroscience, as well as potential clinical applications. PMID- 9333562 TI - [Post-cerebrovascular stroke depression]. AB - Post-cerebrovascular accident depression (PCVAD) affects 30 to 50% of hemiplegic patients in the first two years post-CVA, and has major physical and social repercussions. Particularly closely studied since the beginning of the eighties, PCVAD is considered a therapeutic entity in its own right by many authors. The clinical picture is one of melancholia in 5 to 25% of cases, and of minor or masked depression (with psychomotor retardation and somatic disorders predominating) in 75 to 95% of cases. Etiopathogenesis varies depending on post CVA period: during the first few months, the depletion of intra-cerebral neurotransmitters is considered to play a dominant role; subsequently, difficulty in coping with the handicap would appear to be the main factor. The diagnostic scales which may be used are CIM 10 or DSM IV. For quantification, Hamilton's, the MADRS, Zung's or the CESD scales may be used. There is as yet no scale specific to PCVAD. The therapeutic approach still remains empirical, due to the rarity of published studies. Tricyclic antidepressants and inadvisable as first line treatment due to their anticholinergic effects. Serotoninergic agents are well tolerated, but their efficacy is currently insufficiently documented, despite a recent study. Electroconvulsive therapy (ECT) has been tried, with a certain degree of success, by some authors, but no controlled study is currently available. Personal and familial psychological management would appear necessary but this has not yet been validated. In a preliminary, open-label study in 15 patients presenting with PCVAD, the authors obtained normalization of the MADRS in 10 cases following 6 weeks of treatment with fluoxetine (Prozac). No adverse effects were observed. A multicenter, controlled study is currently ongoing in Bordeaux, France. PMID- 9333563 TI - [The central nervous system, depression and cardiac death]. AB - There is now convincing evidence of a link between depression and cardiovascular death. This evidence comes from prospective studies of healthy populations, and from studies of patients with ischaemic heart disease. It has recently become clear that there is a strong association between smoking and depression. This gave rise to the hypothesis that the association between cardiovascular disease and depression was an artefact due to smoking. The National Health Examination Follow-up Study (NHEFS) settled the question. Three thousand adults without clinical disease at the start of the study were followed up for 12.5 years. After controlling for smoking and other known risk factors, depression was found to be a predictive factor for an increased risk of fatal and non-fatal ischaemic heart disease. Camey et al. in 1988 studied patients rather than healthy subjects, and showed that among patients undergoing coronary angiography, those suffering from major depression had a higher risk of myocardial infarction and of death during the next 12 months than non-depressed patients. Two years later, Ahern et al, found that patients presenting with ventricular arrhythmias following myocardial infarction had a higher risk of death during the following year if they suffered from depression. However, the best study by far is that of Frasure-Smith et al. (1993). They followed up 222 consecutive patients who had suffered a myocardial infarction, and demonstrated that the development of major depression was a significant predictive factor for death during the following 6 months (adjusted risk ratio: 4.29). By the end of 18 months, those patients who had only a slight elevation of the Beck score in the intensive care unit had the same risk of death as those showing symptoms of major depression. As regards the question of whether treatment for depression can alter the risk of death in patients suffering from ischaemic heart disease, and what are the mechanisms underlying these risks, the jury is still out. PMID- 9333564 TI - [Psychopathology and cognitive neurosciences: theoretical conflicts and experimental paradigms]. AB - Clinical research may be seen in the context of the more general problem of an exchange of knowledge between psychopathology and neurosciences. While this interaction is not contested by the specialists in either field, we cannot, nonetheless, ignore the question of their various levels of congruence. More generally, we are increasingly aware of a need for enhanced formalization of the models derived from the two disciplines with a view to setting up transpositions or even cross fertilizations. The two communities, that of psychiatrists and that of neurobiologists, while they fascinate each other, have rarely been able to produce joint findings. The psychopathologist expects a positive validation of his clinical concepts from the neurobiologist, while the neurobiologist, who has frequently not resisted the etiological temptation, wants the clinician to give him pointers to enable him to refine his own experimental models. What is required, however, is that both the psychopathologist and the neurobiologist reduce their expectations and agree to move outside of their respective disciplinary autarkies. This would avoid two traps: that of the homological temptation which leads both to graft an experimental model onto clinical entities without checking the levels of congruence or non-congruence, making do with a simple behavioral similarity; the second error consists in using the complexity of the clinical picture as an unavoidable obstacle to any coming-together of the two disciplines. These two traps have not always been avoided, in particular in the context of the use of cognitive paradigms in nosographic clinical models. PMID- 9333565 TI - Proceedings of the 5th International Symposium on Neurobehavioral Methods and Effects in Occupational and Environmental Health. Cairo, Egypt, December 3-7, 1994. PMID- 9333566 TI - Pertussis Vaccine Trials. Symposium proceedings. Rome, Italy, October 30-November 1, 1995. PMID- 9333567 TI - Pertussis vaccine trials. Trial synopses. PMID- 9333568 TI - The Gothenburg pertussis vaccine study. PMID- 9333569 TI - Efficacy trial of acellular pertussis vaccines; trial I. PMID- 9333570 TI - Italian trial on acellular pertussis vaccines. PMID- 9333571 TI - A comparative efficacy trial in Germany in which infants received either the Lederle/Takeda acellular pertussis component DTP (DTaP) vaccine, the Lederle whole-cell component DTP (DTP) vaccine or DT vaccine. PMID- 9333572 TI - Senegal pertussis trial. PMID- 9333573 TI - Efficacy of a three-component acellular pertussis vaccine (DTaP) in early childhood after household exposure to "typical" (WHO-defined) pertussis. PMID- 9333575 TI - Proceedings and abstracts of the 29th European Pancreatic Club Meeting, Young Researcher's Corner. London, United Kingdom, July 9-12, 1997. PMID- 9333574 TI - Case-control study to evaluate the efficacy of the BIKEN acellular pertussis vaccine combined with diphtheria and tetanus toxoids (DTaP) in infants. PMID- 9333576 TI - [Changes in prenatal diagnosis. Hope for many couples--ethical problems unavoidable]. PMID- 9333577 TI - [Prenatal diagnosis: methods--indications--risks]. AB - The demands made on prenatal care in terms of fetal anomaly recognition and full and early counseling have become more and more sophisticated over the last few years. Today, a range of non-invasive and invasive procedures are now available for the detection of fetal anomalies, genetic disorders and e.g. infection. The most important of the former are ultrasound and triple test (determination of alpha feto-protein, HCG and unconjugated estriol), while amniocentesis remains the major invasive measure. The indications, the information they provide, and possible risks of these procedures are discussed. PMID- 9333578 TI - [Premature labor--central problem of modern obstetrics]. PMID- 9333579 TI - [Optimal prenatal care with the latest techniques]. PMID- 9333580 TI - [Chronic hepatitis. Differential diagnosis and therapy--1]. PMID- 9333581 TI - [AT1 receptor antagonists in treatment of hypertension]. PMID- 9333582 TI - [Opiate dependent patients in inpatient detoxification. Prevalence of somatic diseases in hospital admissions]. AB - AIM: To investigate whether the impression that prevalence rates of diseases in drug addicts undergoing detoxification could be verified empirically. In addition, to make an attempt to identify possible relationships responsible for this development. METHOD: Between 1989 and 1993, within the framework of a detoxification program, 1041 opiate addicts were investigated for the presence of somatic diseases. For this purpose, the overall sample was divided up into 3 groups corresponding to the following time periods: 1989/1990, 1991/1992, 1993. The baseline data comprised the standardised history at the time of admission, the results of the initial clinical examination, and the serological and clinico chemical findings during the further course. RESULTS AND INTERPRETATION: A deterioration in the general state of health associated with more risky drug consumption patterns and social isolation was established. Apparently, low threshold treatment programs and prophylactic strategies made available to almost two-thirds of the addicts in Hamburg have proved unable to prevent the deterioration of the situation of a subpopulation of addicts. PMID- 9333583 TI - [Homocysteine and coronary heart disease. Is slight or moderate homocysteinemia related to increased risk of coronary heart disease?]. AB - Homocystinuria, a rare inherited disturbance of amino acid metabolism is associated with severe atherosclerosis and thromboembolism already in childhood. The incidence of the homozygous disease of cystathionine beta synthase is estimated to be 1:200,000, that of the heterozygous form 1:300. There are, however, numerous other causes of a mild to moderate homocysteinemia, for example, a deficiency of the cofactors vitamin B6, B12 and folic acid. The question as to whether a mild to moderate elevation of homocysteine in the plasma is also associated with an increased risk of CAD has been investigated in a number of studies in recent years. In summary, however, the presently available data do not provide a basis of proof that a mild to moderate homocysteinemia is an independent risk factor for CAD. Our present knowledge suggests that only when the family history is clearly positive does it appear reasonable to undertake a diagnostic search for possible homocysteinemia. PMID- 9333584 TI - [Chronic hepatitis. Differential diagnosis and therapy. 2: Differential hepatitis C diagnosis]. PMID- 9333585 TI - [A rare cause of gynecomastia in a more than 70-year-old man. Case report and differential diagnostic considerations]. AB - Gynecomastia in elderly men is relatively common and may be caused by age-related endocrinal and metabolic disorders, the use of certain drugs, or specific medical conditions. We report here on the rare case of a 77-year-old man in whom gynecomastia was one of the major symptoms of a bronchial carcinoma which had metastasized to the mediastinum. The differential diagnosis of the condition is described in detail. PMID- 9333586 TI - [Mg(2+) transport in the heart. An overview]. PMID- 9333587 TI - [Effect of changed extracellular K(+) and Mg(2+)-concentration on intracellular Ca(2+) homeostasis, contraction coupling and force-frequency relations in the human myocardium]. AB - The intracellular Ca(2+)-homeostasis may be affected by changes of the extracellular K(+)- and/or Mg(2+)-concentrations. Mg2+ reduces the Ca(2+)-influx via L-type Ca(2+)-channels, facilitates Ca(2+)-uptake into the sarcoplasmic reticulum, modulates the Ca(2+)-induced Ca(2+)-release and the Ca(2+)-binding to troponin C. The extracellular K+ activates the Na+/K(+)-ATPase and changes the membrane potential thereby affecting the mode of action of the Na+/Ca(2+) exchanger. Especially when intracellular Ca2+ regulation is altered, for example in heart failure, Mg2+ and K+ exert beneficial effects on the frequency-dependent force-generation in human myocardium. Thus, extracellular Mg2+ and K+ influence contraction coupling in the human myocardium. PMID- 9333588 TI - [Electrophysiologic effects of K+ and Mg2+ in the hypoxia model]. AB - The influence of Mg2+ and K+ on reoxygenation arrhythmias following 18 min of hypoxia (pO2 about 1 mm Hg, no glucose, 10 mmol/l2-desoxyglucose) has been investigated in isolated guinea-pig left atria (stimulation rate 1 Hz). Duration of reoxygenation arrhythmias was slightly reduced by increase in Mg2+ (0.6 to 4.8 mmol/l) and enhanced by decrease in Mg2+ (0.1 mmol/l), however, this effect was not significantly different from control (0.6 mmol/l). In contrast, low K+ (< 4 mmol/l) led to a significant (p < or = 0.05) prolongation and high K+ (> 5 mmol/l) to a significant abbreviation of reoxygenation arrhythmias. Increase in Mg2+ significantly attenuated the proarrhythmic effects of low K+ and enhanced the antiarrhythmic effects of high K+. Furthermore, the influence of Mg2+ and K+ on action potential parameters has been investigated in hypoxic guinea-pig papillary muscles (pO2 65 to 75 mm Hg). Action potential duration at 30% (APD30) and 90% repolarization (APD90) were increased by both electrolytes whereas resting potential, amplitude of the action potential and maximum upstroke velocity were not changed with the exception of a depolarization induced by elevated K+. 1 mmol/l Mg2+ increased APD30 to 145 +/- 16% (n = 6, p < 0.05) and APD90 to 117 +/- 9% (n = 6, p > 0.05) of control (0.6 mmol/l Mg2+). Increase of K+ from 2 mmol/l (control) to 4.7 mmol/l increased APD30 to 188 +/- 13% (n = 6, p < 0.05) and APD90 to 136 +/- 13% (n = 6, p < 0.05). The delay in repolarization observed already in therapeutic concentrations showed no inverse use dependence as it was not attenuated if stimulation rate was increased from 0.17 to 1 Hz which is in contrast to the effects of class 3 antiarrhythmic drugs (for example sotalol). Increase in concentration of both electrolytes led to an additive increase in action potential duration. Mg2+ (0.6 to 4.8 mmol/l) suppressed late afterdepolarizations and -contractions in K(+)-depolarized guinea-pig papillary muscles (27 mmol/l K+, 0.5 mmol/l Ba2+) induced by 2 x 10(-8) mol/l isoprenaline. The change in the triphasic contraction cycle by elevation of Mg2+ indicates that Mg2+ additionally increases stimulus-induced release of Ca2+ from the sarcoplasmic reticulum and reduces slow Ca2+ inward current. The described electrophysiological actions of the electrolytes represent mechanisms, which may explain their antiarrhythmic actions observed in clinical studies. PMID- 9333589 TI - [Comments on the kinetics and extracellular effects of potassium and magnesium]. AB - Potassium and magnesium are important electrolytes which have to be ingested in sufficient amounts. They differ in the necessary daily intake (about 100 mmol potassium, about 12 mmol magnesium), the degree of intestinal absorption (potassium almost 100%, magnesium about 30%) and the distribution between the extracellular and intracellular space. If there is a deficiency in potassium or magnesium, it is necessary to substitute these materials. Deficiency of potassium is not rarely combined with deficiency of magnesium. The concentration gradient between intra- and extracellular potassium mainly determines the resting membrane potential of the cell. Lower extracellular potassium may lead to an instable membrane potential because of a decrease in potassium conductance. An increase in extracellular potassium concentration leads to the depolarization of the cell. Extracellular potassium activates the sodium potassium pump and thereby prevents an increased intracellular accumulation of sodium and calcium. Important effects of extracellular magnesium are: calcium antagonism, increase of excitation threshold and inhibition of transmitter release. By increasing the plasma concentration of magnesium, it is possible to exert pharmacodynamic effects. An increase above the normal range usually is only possible by parenteral application. However, a slight elevation can already be achieved by oral application. This increase may lead to limited pharmacodynamic effects. By elevation of the extracellular magnesium concentration, adverse, depolarisation dependent effects of an increase in extracellular potassium concentration (for example slowing of conduction of excitation) can be compensated. This effect can be explained by a magnesium-dependent decrease of the membrane surface potential. PMID- 9333590 TI - [Effect of magnesium on infarct size after coronary occlusion. Animal experiment studies]. AB - In addition to its antiarrhythmic and antithrombotic effects magnesium is said to have a beneficial effect in patients with acute myocardial infarction. Magnesium can protect myocardial tissue after coronary occlusion and reduces infarct size in experimental models of ischaemia and reperfusion, though the given doses of magnesium are relatively high and differ from clinically reachable serum concentrations. We tested 2 hypotheses in a dog model of ischaemia-reperfusion: 1. The protective effect may be due to a direct, local influence of magnesium on myocardial reperfusion injury. 2. Systemic magnesium treatment with low doses comparable to clinical study regiments may reduce myocardial infarct size. Anaesthetized open chest dogs underwent 1 h of left anterior descending artery occlusion followed by 6 h of reperfusion. 1. Ten animals received intracoronary magnesium aspartate (Mg i.c.) or vehicle infusion (control i.c.) for the first hour of reperfusion to increase regional Mg-concentration by 2 mmol/l. 2. Fourteen animals received intravenous infusion with magnesium potassium aspartate (Mg-K i.v.) or vehicle infusion (control i.v.), beginning 1 h before occlusion until the end of the 6 h reperfusion period. Regional magnesium concentration in the Mg i.c.-group increased to 2.7 +/- 1.00 mmol/l at 45 min of reperfusion. Intravenous infusion raised serum magnesium from 0.71 +/- 0.03 mmol/l to 1.29 +/- 0.14 mmol/l in the Mg-K i.v. group (5 min of reperfusion, p < 0.01 vs. baseline). Infarct size after 6 h reperfusion (TTC staining) was similar in both groups of intracoronary treatment (Mg i.c., 20.6 +/- 5.0; control, 24.4 +/- 8.7% of area at risk; p = n.s.) and intravenous treatment (Mg-K i.v. 18.1 +/- 14.8; control 14.1 +/- 12.2% of area at risk; p = n.s.). Neither regional nor systemic magnesium leads to a clinically relevant reduction of infarct size in the regional ischaemic-reperfused dog heart when it is given in clinically usable doses. The beneficial action of systemic Mg is probably not due to an early direct protective effect on ischaemic-reperfused myocardium but to its antiarrhythmic and antithrombotic effects. Possibly only to high doses of Mg applied under experimental conditions can reduce infarct size. PMID- 9333591 TI - [Clinico-electrophysiologic effects of magnesium, especially in supraventricular tachycardia]. AB - Clinical electrophysiological effects of magnesium (Mg2+) are known for more than 60 years. Mg2+ is a cation to be found ubiquitously in the human body and is involved in more than 300 different enzymatic reactions. However, so far this ion has not been established as a standard therapeutic tool for the treatment of supraventricular tachyarrhythmia. This may be explained by the inconsistent efficacy of Mg2+, partly in relationship to a given plasma Mg(2+)-concentration, partly caused by the uncertainty regarding the dosage and injection rate or the unawareness of the clinical effects of the cation. Mg2+ influences myocardial metabolism by its effects on contractility and electrical activity. Both effects are closely linked. About 12% of cardiac Mg2+ is found in the mitochondria and 2 to 3% in the myofibrils. A large portion is incorporated in adenosin mono-, di- and triphosphate. Mg2+ affects intracellular calcium by inhibiting the influx of calcium into the myocyte through sarcolemmal channels, by modulation of cyclic AMP and by competing with calcium for binding to a single high affinity site on actin. Mg2+ has been linked to a naturally occurring calcium channel blocker. Furthermore Mg2+ blocks the outward current through some potassium channels resulting in an inward rectification of these channels. This suggests that internal magnesium functions as a potassium channel-blocking agent. Early afterdepolarizations are oscillations in the membrane potential and lead to triggered activity and therefore are the electrophysiological substrate of "torsade de pointes" type of ventricular flutter. Mg2+ is able to inhibit both early afterdepolarizations and tachyarrhythmias. Additionally Mg2+ interferes with the sodium-potassium-ATPase system by stabilizing the transmembrane gradient of both cations. Mg2+ deficiency alters this balance and leads to increased neuromuscular excitability. Digitalis is able to block the sodium-potassium ATPase system, which can be cancelled by Mg2+. Thus the first clinical reports of the therapeutic use of Mg2+ refer to digitalis-induced atrial arrhythmia and ventricular ectopy which could be converted to sinus-rhythm or suppressed by the intravenous application of Mg2+ in 1935. Some years later, the first successful termination of paroxysmal supraventricular and ventricular tachycardia following application of 1.5 to 3 g of Mg2+ was published. But only in the late eighties, systematic studies of the electrophysiological effects of Mg2+ were performed and clinical use was first tested in random fashion in the nineties. Summarizing studies in older patients with different heart diseases and young healthy volunteers the most pronounced and clinically important effect seems to be related to the modulation of the AV node function. The prolongation of the PR interval by 7 to 12% without changing significantly heart rate, QRS duration and QT duration, can be considered a consistent and reproducible effect of Mg2+. In electrophysiological studies a prolongation of the AH interval by 8 to 18%, of the Wenckebach cycle length by up to 20% and of the refractory period of the AV node by 6 to 20% is usually observed, but no change of the retrograde conduction, or the HV interval can be found. Furthermore sinus node recovery time increases by 10% and sinuatrial conduction time by up to 25%. There is no significant effect on intraventricular conduction and atrial and ventricular refractory period. Additionally no significant effect on the anterograde and retrograde refractory period of accessory pathways could be measured; however in some cases (up to 40%) an anterograde block in the accessory pathway may be observed after intravenous Mg(2+)-injection. For the treatment of paroxysmal atrioventricular tachycardia like AV-nodal reentrant tachycardia or orthodromic atrioventricular reentrant tachycardia in WPW syndrome, Mg2+ has been applied in a limited number of recent prospective but uncontrolled studies. Recently, an PMID- 9333592 TI - [Effect of magnesium on sustained ventricular tachycardia]. AB - Intravenous application of magnesium was suggested for the management of persistent ventricular tachycardia. In patients with torsade de pointes tachycardia the injection of magnesium controlled the life threatening arrhythmias reliably, thus magnesium became the treatment of choice in this setting. The results in patients with persistent monomorphic ventricular tachycardia are controversial. In one study, ventricular tachycardias could be controlled in 8 of 10 patients by 2,000 mg magnesium sulfate. In a randomized trial with 43 patients, termination of the ventricular tachycardia could be accomplished in 6 of 20 patients under the influence of 4,000 mg magnesium sulfate, whereas 3 of 24 patients exhibited termination of the tachycardia following placebo; the difference did not reach statistical significance. In our study 4 patients with torsade de pointes tachycardia could be controlled by the application of magnesium glutamate (2 x 1,000 mg intravenously). Of 25 patients with persistent monomorphic ventricular tachycardia, the arrhythmia ended in 8 patients under the influence of magnesium. No significant change of the RR intervals and the QRS duration during ventricular tachycardia could be demonstrated following magnesium injection. Cardiac index during ventricular tachycardia increased from 2.0 +/- 0.6 l/min x m2 to 2.5 +/- 0.1 l/min x m2 (p < 0.05). In a further investigation the dose of magnesium was increased to 2 x 9 mmol. To detect possible interactions with antiarrhythmic agent, 10 patients under chronic antiarrhythmic therapy with class-III-agents were compared with 10 patients without such treatment. Plasma levels of magnesium increased from 0.79 +/- 0.1 mmol/l to 1.87 +/- 0.5 mmol/l. In 5 of the 20 patients ventricular tachycardia ended under the influence of magnesium: 2 patients were on chronic antiarrhythmic agents, whereas 3 had no chronic therapy. There were no significant differences in the RR intervals and the duration of the monophasic action potentials during the ventricular tachycardia under the influence of magnesium. These data indicate that the bolus therapy of magnesium controls persistent monomorphic ventricular tachycardia in a minority of patients only. Therefore, magnesium injection cannot be recommended for treatment of monomorphic ventricular tachycardia in the emergency setting. On the other hand, magnesium application can be considered the therapy of choice for patients with torsade de pointes tachycardia. PMID- 9333596 TI - Invited contributions from the 13th European Immunology Meeting. Amsterdam, The Netherlands, 22-25 June 1997. PMID- 9333593 TI - [Magnesium deficiency and magnesium substitution. Effect on ventricular cardiac arrhythmias of various etiology]. AB - During recent years there has been an increasing but still controversial discussion on the antiarrhythmic effects and overall benefit of magnesium when directed to patients with various types of ventricular tachyarrhythmias. While magnesium is considered to be a simple, safe and cost-effective approach and many casuistic and empiric reports have indicated antiarrhythmic properties of magnesium in patients with suspected or manifest ventricular arrhythmias, controlled studies proving the antiarrhythmic and overall benefit and justifying a broader use of magnesium in treating various types of ventricular arrhythmias are missing or rare. At present, antiarrhythmic properties and clinical benefit of magnesium application has only been established in patients with torsade de pointes and digitalis-induced ventricular tachyarrhythmias. In perioperative patients at risk for ventricular tachyarrhythmias and in patients suffering from manifest heart failure, data may also indicate some antiarrhythmic properties of magnesium, however, in this case with a wide consensus that the prevention of magnesium deficit is more effective and preferred in most patients over the therapeutic application of magnesium. Another group of patients who may profit from such a therapeutic approach are patients with frequent ventricular arrhythmias and stable underlying heart disease, in whom a recently published double-blind, randomized study documented an antiarrhythmic effect of a 3 week treatment with potassium and magnesium. For all other types of ventricular tachyarrhythmias, the therapeutic use of magnesium can be considered as not harmful, but also as not proven to be effective. PMID- 9333595 TI - [Medicamentous anti-arrhythmia therapy. Is oral adjuvant therapy with electrolytes of value?]. AB - Low serum concentrations of potassium and magnesium are proarrhythmic factors that are well established. Atrial and ventricular fibrillation are facilitated at low serum levels of these electrolytes. Loss of potassium and magnesium might be caused by diuretic therapy, gastrointestinal loss, drugs, and alcohol abuse. However, serum levels are not representative of total body content of potassium and magnesium, hence, adjuvant therapy might be indicated also in the presence of normal serum levels. This is especially true during the initial phase of antiarrhythmic therapy, which is accompanied by proarrhythmia in a significant number of cases. Patients with heart failure should routinely receive adjuvant electrolyte substitution, if renal function is not impaired. In the experimental model magnesium successfully prevented early afterdepolarizations caused by hypokalemia and antiarrhythmic drugs. In the clinical setting high dose magnesium abolished torsade-de-pointes tachycardias caused by antiarrhythmic drugs. Unfortunately, controlled studies are not available for low dose electrolyte therapy adjuvant to antiarrhythmic drug medication. PMID- 9333594 TI - [Value of K+ and Mg2+ in treatment of acute myocardial infarct]. AB - A critical role analysis of literature concerning the effects of intravenous magnesium on arrhythmias and mortality in acute myocardial infarction shows discrepant results and often inappropriate methods. So far neither an antiarrhythmic efficacy nor prophylactic effects with respect to mortality could be demonstrated. In contrast, potassium substitution should be performed in the setting of acute myocardial infarction with documented hypokalemia (K+ < 3.5 mmol/l) because of increased risk of ventricular arrhythmias. According to the documented results of the trials reviewed in this article no recommendations for the routine use of magnesium in myocardial infarction can be given. PMID- 9333597 TI - [Inhibition of metastases by antithrombotic agents]. PMID- 9333598 TI - [Thrombophilia and antithrombotic prevention in gynecology and obstetrics]. PMID- 9333599 TI - [Antithrombotic prevention and therapy in coronary heart disease]. PMID- 9333600 TI - [Prevention with low dosage heparin. Selective or general?]. PMID- 9333601 TI - [Indications and problems of long-term medicamentous inhibition of blood coagulation]. PMID- 9333602 TI - [New antithrombotic drugs. What developments can be expected?]. PMID- 9333603 TI - [Temporary hemiparesis and Raynaud symptoms in eosinophilic fasciitis]. PMID- 9333604 TI - [Liver transplantation in familial amyloid polyneuropathy. Case report and review of the literature]. AB - A 59-year old male of German origin noticed exercise-independent cardiac arrhythmia two years before admission. An alanine 47 transthyretin variant of Familial Amyloid Polyneuropathy with hypertrophic cardiomyopathy, peripheral sensory-motor polyneuropathy, I, degree AV heart block was diagnosed. To diminish production and deposition of mutant transthyretin and to prevent disease progression orthotopic liver transplantation was performed. Prior to transplant the patient complained of inappetence. Postoperatively, he received a chemically defined enteral nutrition regime that was discontinued after 30 months until return of appetite and weight gain indicated marked improvement. However, a duodenal biopsy still demonstrated amyloid deposits 24 months after transplantation. Echocardiographic findings remained unchanged. Neurologic examination showed an improvement of sensory-motor polyneuropathy with regression of electromyographic changes. Only traces of variant transthyretin were detectable in plasma samples taken 12 months after the operation. During the 3 year follow-up, no additional symptoms have occurred and progression of amyloidosis was prevented. Currently, orthotopic liver transplantation is the only specific treatment to prevent progression of familial amyloid polyneuropathy. PMID- 9333605 TI - [Pathophysiology of Av nodal reentrant tachycardia]. PMID- 9333606 TI - [Treatment of sleep disorders]. PMID- 9333608 TI - [Vaccinations in multiple sclerosis]. PMID- 9333607 TI - [Vitamin supplementation]. PMID- 9333609 TI - [Paracetamol--the safe analgesic]. PMID- 9333610 TI - [Anorexia and bulimia nervosa]. PMID- 9333611 TI - [Dermatology and the Internet--uses for the clinic and research]. AB - The Internet is about to become the most important source of information in virtually any academic profession. Dermatology has a growing presence in many areas of the Internet: discussion groups and continuing education programs invite regular participation, dermatologic data in the form of text and images provide support in daily clinical and scientific work. In this paper important sources of dermatologically relevant information on the Internet are given, and the Dermatologic Online Image Atlas (DOIA) which is available on the World Wide Web is introduced. PMID- 9333612 TI - [Treatment of atopic eczema in childhood]. AB - Adjuvant basic therapy plays a central role in the therapeutic approach to atopic eczema in childhood. It involves the regular use of emollients and oil baths according to the clinical picture. Emollients containing urea may cause problems when used in younger children due to their stinging effect. Glucocorticosteroids still form the mainstay of anti-inflammatory therapy and are superior to other topical drugs with anti-inflammatory effects. Antihistamines preferably of the sedative type can be used successfully during periods of exacerbation. Systemic use of antibiotics is essential in cases of impetiginized atopic eczema. In cases of bacterial-triggered eczema, the topical use of antibiotics in cases of limited or antiseptics in generalized disease may be helpful. PMID- 9333614 TI - [Prevention of melanoma by sun protective measures in childhood. Temporal changes in awareness of parents]. AB - Numerous epidemiological studies on risk factors of malignant melanoma confirm the etiologic role of excessive UV-exposure especially in childhood. Preventive educational campaigns directed to parents of pre-school children have been inaugurated in several countries. In Germany the information was distributed by the "Working group for Preventive Measures in Dermatology" in cooperation with different public health institutions and the media starting in 1993. To evaluate the influence of these efforts on the knowledge and behaviour of the parents, two successive cross-sectional studies at all 56 nursery schools using the same standardised questionnaire were performed. The first interview took place in spring 1993 (before the campaign) with 1341 evaluable questionnaires', the second in fall 1994 (after the campaign) with 1150 evaluable questionnaire. The knowledge of the parents on melanoma risk factors was significantly improved in the second interview. Also the parental behavior regarding sun-protective measures when their children were outdoor at the beach or in the garden definitely changed. In 1993 the best textile sun protection was used by 21% of the parents at the beach and 36% in the garden. These numbers rose to 34% (beach) and 57% (garden) by the second interview. The percentage of children with no sunburn recorded during the preceding summer rose from 39% to 51%. According to the child's gender the parental behavior was different between the sexes; boys were always better protected than girls. The design of this study with two cross sectional surveys in the same populations does not provide a methodologically sound basis for attributing the observed positive changes to the campaign. Without any doubt it can be stated that the parental knowledge and their attention to sun protection in their children showed substantial improvement in the second survey after the campaign. Thus, these results provide some evidence for the success of the preventive activities and confirm the necessity to continue with such activities. PMID- 9333613 TI - [Vascular sports in ambulatory therapy of venous circulatory disorders of the legs. Diagnostic, therapeutic and prognostic aspects]. AB - In 33 patients with chronic venous incompetence (CVI) caused by primary varicoses or postthrombotic syndrome stage I-III (according to Widmer) the therapeutic benefit of 6 months of medically supervised physical exercise training was documented. During the training penud there was an improvement in subjective complains such as pain and tendency for edema in the legs. Mobility in the upper ankle joint was improved asuss as venous drainage function. Clinical benefit was achieved in the reduction of ulcer size; 7 of the 10 ulcers completely healed. Medically supervised physical exercise training and optimized compression therapy are basic therapeutic approaches in conservative treatment in chronic venous insufficiency. Costs are covered by the patient's health insurance company in Germany, as long as the exercise training is medically supervised. PMID- 9333615 TI - [Effect of domestic laundry processes on mycotic contamination of textiles]. AB - Inadequately decontaminated clothing may be a source of reinfection following therapy of dermato- and onychomycoses. The objective of this study was to determine whether domestic laundering is suitable for cleansing mycotically contaminated garments. Textile-samples contaminated with Trichophyton rubrum, Trichophyton mentagrophytes, Candida albicans and Scopulariopsis brevicaulis were washed in an ordinary washing machine at different temperatures. Regardless of the textiles and detergents used, reliable decontamination was achieved by laundering at 60 degrees C. Trichophyton rubrum was eliminated with a washing temperature of 30 degrees C. PMID- 9333616 TI - [Granuloma pyogenicum--removal with the CO2 laser]. AB - 13 patients with pyogenic granuloma were treated with a CO2 laser using the continuous wave laser in all cases and using additionally the ultrapulsed laser in 7 patients. The pyogenic granuloma was removed in all patients in one session. Side effects included transient erythema in 8 patients and scars in 2 cases. There was no hypo- or hyperpigmentation. The use of the CO2 laser is a fast and bloodless way to treat pyogenic granuloma with slight side effects and is an elegant alternative to standard approaches. PMID- 9333617 TI - [Prurigo pigmentosa]. AB - We report the first case of prurigo pigmentosa in the German literature. For over a year, a 16-year old white girl developed an immensely pruritic symmetrical reticular, papulovesicular eruption on the trunk, axillae and pubic region. Histologically a superficial perivascular spongiotic and lichenoid inflammatory pattern with prominent pigmentary incontinence in older lesions was observed. Topical corticosteroids and systemic antihistamines had no effect. Systemic corticosteroids and diaminophenylsulphone (DADPS) produced improvement but could not prevent recurrences. Healing occurred only after 4 weeks of systemic minocycline. PMID- 9333618 TI - [Lichenoid pseudolymphomatous tattooing reaction]. AB - A 35 year old patient developed swellings in the red colored areas of his tattoo. Histological examination revealed a lichenoid, pseudolymphomatous infiltrative pattern, that could be distinguished from frank lymphoma by means of electron microscopy, immunohistochemistry and molecular biology. The presence of dermal dendritic cells suggests a dermal-allergic pathogenesis of non-granulomatous tattoo reactions. Therapy of choice is an excision of the inflamed areas. PMID- 9333619 TI - [Torus mandibularis]. AB - Solitary or bilateral, symptomless exostoses on the lingual surface of the mandibule are called mandibular torus. It is mainly seen in young males and has a benign clinical course. The etiopathology is not known. Both genetic and environmental factors such as the anatomy of the lower jaw are considered. Syndromes associated with facial exostoses such as Proteus syndrome or Gardner's syndrome should be clinically excluded. A 40-year-old man with exostoses of the jaw is reported. With this case report we would like to draw attention to a disease which has rarely been described in the German dermatological literature. PMID- 9333620 TI - [Adenoid cystic cribriform trichoblastoma]. AB - In hair germ tumors, hair follicle development is partially or completely recapitulated. The trichoblastoma represents a purely epithelial tumour within this group of tumours. We identified a trichoblastoma showing exclusively adenocystic features in a 32 years old woman. This variant has been very rarely described in the literature. We review the history of hair germ tumors and discuss the problems of their classification. PMID- 9333621 TI - [Atrophic polychondritis with multiple arterial occlusions]. AB - A 68-years old patient with relapsing polychondritis of the ear developed stenosis and occlusion of hercarotid and femoropopliteal arteries. Relapsing polychondritis is a rare systemic disorder which may take a fetal course. We also review this rare, systemic disorder. PMID- 9333622 TI - [50 years Zwickau Dermatologic Clinic in the Heinrich-Braum Hospital]. AB - The dermatological clinic in Zwickau celebrated 50 years of existence in 1996. We review the clinics history, starting with its founding in 1946 because of the upsurge in venerological diseases during the post-world war II years. The most important clinic directory and their scientific activities are described. PMID- 9333623 TI - [Imaging methods in dermatology]. PMID- 9333624 TI - [Brittle nails. Objective assessment and therapy follow-up]. AB - The different methods that are available for the assessment and quantification of "brittle nails" and for the evaluation of various therapeutical approaches are compared. At this time, measurement of the swelling properties is the best documented and most reliable method for studying the treatment of brittle nails. Reliable qualitative data can also be obtained by scanning electron microscopy. Measurement of the transonychial water loss and assessment of thickness and density of nails by ultrasound have also been used successfully, but the methodology still has to be improved and the reproducibility, confirmed. PMID- 9333625 TI - [Q-switched ruby laser in dermatologic therapy. Use and indications]. AB - The Q-switched ruby laser (QSRL) with its wavelength of 694 nm and a pulse duration of around 40 nsec is an effective modality for the removal of tattoos and cutaneous pigmented lesions. Based on the principle of selective photothermolysis, selective damage to cutaneous pigment or pigmented cells is possible, allowing the scar-free elimination of endogenous or exogenous pigment in the skin. Main indications for the treatment with the QSRL are tattoos (amateur, professional, accidental, or cosmetic) and lentigines but the QSRL can also be used for lightening or even removing other pigmented lesions such as nevus spilus or cafe au lait macules. Furthermore, pigmented lesions of mucous membranes can be removed easily. Since treatment results in postinflammatory hyperpigmentation, myoplasma, and Becker' nevus have proven to be inconsistent, the QSRL cannot be routinely recommended for these lesions. Melanocytic lesions are generally not treated, with the exception of nevus of Ota and nevus of ito where there exacts a lack of therapeutic alternatives. Non-pigmented cells, which exist in nearly all melanocytic lesions, do not absorb the light of the QSRL and, therefore, do not react to this particular treatment. No information is available on the risk of partially damaged cells to become malignant after QSRL treatment. The QSRL is an excellent therapy for the removal of endogenous and exogenous pigment because of both the excellent treatment results and the lack of side effects, which are limited to transient hypo- and hyperpigmentation. The QSRL has occurred a wide range of applications within the field of dermatology. PMID- 9333626 TI - [Dynamic in vivo skin pressure measurement in quality control of compression stockings]. AB - The well-documented positive effect of compression stocking therapy on the venous macro- and microhemodynamics of the legs can only be attained if the stockings fit well. In order to determine the effective pressure exerted by compression stockings, we usually deleted in US journals. One can get this out of journal and author's address have developed a new measuring method based on piezoresistant microprobes and a microprocessor unit. With our 2-mm-thick, 5-mm diameter probe, the pressure between the compression stocking and skin can be measured at any location desired. A temporal resolution of 50 Hz makes it possible to carry out dynamic measurements while the patient is walking or performing exercises on tiptoes. Here we present 4 typical cases out of a total of over 80 which we have evaluated. We have decided empirically that the pressure exerted by a class-2 compression stocking on the skin at the height of the ankles (b-position) should not exceed 70 mm Hg while resting and a peak of 110 mm Hg while exercising on tiptoes. At the middle of the calf (c-position) these values should not exceed 60 mm Hg at rest and 80 mm Hg on tiptoes. The pressure should decrease from the distal to proximal direction in order to produce a drainage gradient. We have found empirically that a pressure gradient of 30-40% from the b to the c measurement is favorable. Too high a proximal pressure or too high a pressure on a part of the lower leg causes pain and swelling. Too low a pressure, on the other hand, does not produce the desired vascular effect and alleviation of symptoms. Although dynamic pressure measurements take about 20-30 minutes per leg, they markedly improve patient compliance with compression therapy. PMID- 9333627 TI - [Oral DMSO therapy in 3 patients with lipoidproteinosis. Results of long-term therapy]. AB - Lipoid proteinosis is a rare autosomal recessive disorder with a chronic, benign course. There is no generally accepted systemic therapy apart from the experimental oral use of dimethyl sulphoxide (DMSO) and etretinate in two single cases. We treated two sisters and an unrelated man with lipoid proteinosis with longterm oral DMSO (60 mg/kg/d). At the end of an average treatment time of 3 years, DMSO was withdrawn because it produced no beneficial effects with regard to their skin, mucosal lesions or hoarseness. Additionally, one patient showed progression of her disease with worsening hoarseness and onset of dyspnea, requiring surgical removal of vocal cord infiltrates. Three patients with lipoid proteinosis failed to show any beneficial response to long term treatment with DMSO. PMID- 9333629 TI - [Balneophotochemotherapy with 8-methoxypsoralen in lichen sclerosis et atrophicus]. AB - A 66-year-old woman with longstanding lichen sclerosus et atrophicus improved strikingly with PUVA bath photochemotherapy over a period of 6 weeks. The cumulative UVA dose was 31.7 J/cm2; the single UVA dose ranged from 0.3 to 2.3 J/cm2. After 16 treatment sessions, the sclerotic lesions had softened greatly, while after 24 treatments, the skin lesions were almost completely cleared and pruritus was diminished. Histopathological analysis of biopsy specimens from previously affected sites as well as 20 MHz ultrasound examinations showed almost no residual sclerosis. Although long-term results are not yet available, PUVA bath photochemotherapy seems to be a promising and effective new treatment modality without systemic side effects for patients with disseminated lichen sclerosus et atrophicus. PMID- 9333628 TI - [Sarcoidosis in interferon-alpha therapy]. AB - Sarcoidosis is characterised by the formation of sarcoidal granulomas in all affected organs. Despite intensive research, the cause of the disease in unknown. There are only a few reports suggesting an induction of sarcoidosis by interferons. Three patients are presented in whom sarcoidosis developed during or after interferon alpha therapy. The probability of interferon therapy as cause of sarcoidosis is discussed. PMID- 9333630 TI - [Atypical zosteriform segmental embolia cutis medicamentosa]. AB - A 35-year-old male suffered from an extremely painful embolia cutis medicamentosa. The atypical segmental localization first led to the diagnosis of herpes zoster and an antiviral therapy. Regarding the unusual course of the complication following an intramuscular injection in this case, the question for the real pathomechanisms still remains. PMID- 9333631 TI - [Teleangiectasia haemorrhagica hereditaria. Surgical therapy of malignant skin tumors (Osler-Weber-Rendu disease]. AB - A 65 year old male patient was diagnosed with hereditary hemorrhagic telangiectasia at 30. During recent years he has suffered frequent, almost daily, nose bleeds causing anemia and making several blood transfusions necessary. In the past 2-3 years, the patient has developed multiple squamous cell carcinomas on the face. These unusually large tumours were treated by micrographic surgery using paraffin sections and the defects dosed with a variety of flaps and grafts. Several solar keratoses were also removed. If hemostasis parameters are normal, skin surgery can be performed without hesitation in hereditary hemorrhagic telangiectasia. PMID- 9333632 TI - [Psoriatic onycho-pachydermo-periostitis (POPP)]. AB - We describe a patient with painful enlargement and severe nail deformity of both great toes. The patient furthermore presented with some minor diagnostic criteria for psoriasis such as nail pitting and had a positive family history of the disease. In the recent literature, such cases have been recognized as a new entity termed psoriatic onycho-pachydermo-periostitis (POPP). In this case report, POPP is discussed in relationship to other forms of psoriatic arthropathy and with special emphasis regarding its diagnostic criteria and therapeutic implications. PMID- 9333633 TI - [Capillary microscopy in angiokeratoma corporis diffusum]. PMID- 9333634 TI - ["Inflammations of the male adnexa--disorders in fertility?" Report of the IV. Colloquium Andrologicum 15 February 1997 in Jena]. PMID- 9333635 TI - [Stratum corneum]. PMID- 9333636 TI - Acupuncture on animals raises questions. PMID- 9333637 TI - Animal rights, or simply, compassion? PMID- 9333638 TI - Animal rights, or simply, compassion? PMID- 9333639 TI - In defense of pet friendly stores. PMID- 9333640 TI - Thoughts on curriculum reform. PMID- 9333641 TI - The dark side of laughter: males and U.S. health care. PMID- 9333642 TI - Advertising dentistry. PMID- 9333643 TI - Shortage of dentists. PMID- 9333644 TI - Professional ethics. PMID- 9333645 TI - At-home oral irrigation unscientific. PMID- 9333646 TI - Dental anxiety. PMID- 9333648 TI - Consultation section. Post-cataract hyperopia therapy. PMID- 9333649 TI - An esthetic denture clasp for maxillary canine teeth. AB - The clasp was relatively unobtrusive and resembled a small gold restoration (Fig. 1). The use of a noble alloy provided a clasp that was retentive and also resilient with a good yield strength, therefore the likelihood of failure in service was minimal. The process of soldering to a cobalt-chrome framework was simple. Alternatively, the connecting component of the clasp could be incorporated within resin if an acrylic major connector was used. PMID- 9333647 TI - Interactions of viruses with dendritic cells: a double-edged sword. PMID- 9333650 TI - Handles on custom trays for complete denture master impressions. PMID- 9333651 TI - Religious ritual and dissociation in India and Australia. PMID- 9333652 TI - Effects of achievement motivation and study habits on Nigerian secondary school students' academic performance. PMID- 9333653 TI - Assessing family differentiation: four areas of competence in mother-adolescent relationships. PMID- 9333654 TI - Do tests predict malingering in defendants with mental retardation? PMID- 9333655 TI - [Bacterial translocation induces remnant liver injury after subtotal (90%) hepatectomy in rats]. PMID- 9333656 TI - [A case of acute pancreatitis induced by interferon therapy for chronic hepatitis C]. PMID- 9333657 TI - [Experiences with and observations on cataract surgery in highly myopic eyes]. PMID- 9333658 TI - [Angiogenesis and inhibition of angiogenesis in the eye]. AB - BACKGROUND: Angiogenesis is the formation of new blood vessels from preexisting vessels. Angiogenesis plays an important physiologic and pathologic role in the eye. Pathologic angiogenesis can be found in major causes of human blindness as in diabetic retinopathy and age-related-maculopathy. In recent years great progress has been made in recognizing the mechanisms and regulation of angiogenesis. RESULTS: The general mechanism and the regulation of angiogenesis with proliferative diabetic retinopathy as an example of ocular angiogenesis are reviewed according to recent literature. Angiogenic and antiangiogenic factors regulate the neovascularization. Recent research has identified the vascular endothelial growth factor as the most important mediator of ocular angiogenesis. As well in diabetic retinopathy as in age-related-maculopathy the vascular endothelial growth factor plays a role. New knowledge about ocular angiogenesis is linked to the chance of new antiangiogenic therapies in neovascularizing eye diseases. There exist two ways of ocular antiangiogenic therapy: the first way is to block ocular angiogenic factors such as vascular endothelial growth factor, the other way is to influence the interaction between endothelial cells and extracellular matrix of the newly forming vessels. CONCLUSION: Recent progress in angiogenesis research could result in new causal antiangiogenic drug therapy in ophthalmology. There are some promising animal experiments of local and systemic antiangiogenic therapy. Because of its anatomic localisation the eye is especially suitable for topic antiangiogenic therapy. PMID- 9333659 TI - [Use of of the high frequency capsulotomy instrument in cataract surgery--effect of coagulation on corneal endothelium]. AB - Intumescent or hypermature cataracts make a safe capsulorhexis impossible. High frequency capsulotomy represents a satisfying solution for this problem. Primary goal of the present study was to investigate a possible damage to the corneal endothelium by this method. MATERIALS AND METHODS: 55 patients with an uncomplicated senile cataract were enclosed into a prospective randomized study undergoing cataract surgery with capsulorhexis or with high frequency capsulotomy. Corneal endothelium was examined preoperatively as well as postoperatively at several intervals. RESULTS: Concerning loss of endothelial cells and parameters of polymegatism and pleomorphism there were no statistically significant differences between both groups. CONCLUSION: The diathermy during high frequency capsulotomy does not show any clinically relevant negative effects on the corneal endothelium within cataract surgery. PMID- 9333660 TI - [Effect of nifedipine on ocular pulse amplitude in normal pressure glaucoma]. AB - BACKGROUND: Ocular pulse amplitude (OPA) is reduced in normal tension glaucoma (NTG) patients when compared to non-glaucomatous, healthy control subjects. This might be related to a vasospastic reaction. The objective of this study was to determine if low OPA in NTG is associated with a vasospastic reaction and its response to vasodilation. METHODS: Nifedipine, a calcium channel blocker, vasodilator and systemic antihypertensive agent improves visual fields in NTG patients following acute and chronic dosing. The effect of 60 mg of daily orally administered nifedipine on OPA, intraocular pressure (IOP, German abbreviation: IOD), blood pressure (BP, German abbreviation: RR) and pulse rate (PR, German abbreviation: HF) were measured prior to and for 3 months after initiating nifedipine therapy in 32 NTG patients with and without a vasospastic reaction as manifested by a local cold exposure test. Before treatment, all patients had reduced OPA evaluated with the Langham Ocular Blood Flow System. RESULTS: During nifedipine treatment NTG patients with a vasospastic reaction showed a significant (p < 0.001) increase in OPA, whereas NTG patients without a vasospastic reaction showed no sig. (p > 0.05) change in OPA. CONCLUSION: There may be two different subgroups of NTG patients, those who have a vasospastic reaction and react to nifedipine, while others lack the ability to react to nifedipine or might have a different, non-vasospastic pathology. Calcium channel blockers and other vasodilators may be useful in the treatment of vasospastic NTG patients. PMID- 9333661 TI - [Effectiveness of dorzolamide as added therapy in glaucoma patients with maximum tolerated drug therapy. A pilot study]. AB - BACKGROUND: The purpose of this study was to evaluate prospectively the value of dorzolamid as an adjunct to maximum tolerated medical therapy in glaucoma patients in whom surgery would otherwise be required. METHODS: 32 eyes of 21 patients with primary open angle glaucoma (14 patients), glaucoma in aphakia (3 patients), pigmentary glaucoma (2 patients), juvenile glaucoma (1 patient) and exfoliative glaucoma (1 patient) were included. The effect of additional dorzolamid application on intraocular pressure (IOP) was determined after 2 hours, 4 days, 4 weeks, 3 months and 6 months. After 6 months, visual fields were checked. RESULTS: Average reduction of IOP in eyes in which dorzolamid was continued was determined to be 31% after 2 hours, 18.3% after 4 days, 17.9% after 4 weeks, 12.1% after 3 months and 9.7% after 6 months. 19 (59%) eyes continued to receive dorzolamid after 6 months without being operated. No progression of glaucomatous damage could be detected in these eyes. In 11 (34%) eyes, treatment with dorzolamid was discontinued. 2 patients (4 (12%) eyes) did not tolerate local side effects of dorzolamid. In 7 (22%) eyes reduction of IOP was insufficient and filtration surgery had to be performed immediately. CONCLUSION: The results of the present study demonstrate that dorzolamid represents an alternative to an immediate surgical management in patients on maximum tolerated therapy for at least six months. PMID- 9333662 TI - [Relation between relative afferent pupillary defect and suprathreshold automated perimetry]. AB - The purpose of this study was to test for correlation between relative afferent pupillary defect (RAPD) and visual acuity, central differential light threshold and visual field defect measured by suprasthreshold automated grid perimetry. METHODS: In 98 patients with optic neuropathies of different origin (inflammatory, ischemic, or compressive) in addition to a standard ophthalmological examination, static perimetry of the 30 degrees visual field with a dense grid of test spots, including measurement of the central differential light sensitivity, was done by means of the Tubingen Automated Perimeter. RAPD was tested using the swinging flashlight test quantified with neutral density filters. RESULTS: Visual acuity and central differential light sensitivity correlated only weakly with the RAPD. The visual field showed a better correlation (coeff. of corr. for all patients 0.58, for patients with optic neuritis 0.64. for patients with AION 0.53 and for patients with compressive neuropathies 0.84). One test spot not seen in perimetry corresponded to an RAPD of about 0.01 log-units. CONCLUSIONS: The results confirm those from previous studies obtained by threshold perimetry. The RAPD is a good parameter for the severity of an optic nerve lesion, especially of compressive origin. PMID- 9333663 TI - [Eye symptoms in Schimmelpenning-Feuerstein-Mims syndrome, a rare phacomatosis]. AB - BACKGROUND: Children with connatal malformations of the face and of the eyes may run a long diagnostic and narrow therapeutic way. By an early diagnosis, this procedure could become significantly shorter und much easier. PATIENTS HISTOPATHOLOGY: Three clinical observations give typical examples: In correlation to the history of a patient described in 1975 by Meythaler (18) we examined two babies with naevus sebaceus Jadassohn, colobomas of the lids, epibulbar tumors, microphthalmus, and white, vascularized corneal opacifications. Already at the age of four weeks we were able to confirm the diagnosis and documented our findings in anesthesia by tables, photographs, and histopathologic observations. FINDINGS: Localisations and expression of the symptoms are very different in patient 2 and 3. In patient 2 the lid hamartoma contained lacrimal gland, cartilage and bone structures. Reconstructive lid operations corrected the lid function and resulted in esthetic amelioration. CONCLUSIONS: The important points in our casuistics were: 1. After comparison of our findings with those in literature, there was not found any other patient with scalp defect as shown in patient 2. A cataract as described by Piper (21) was not seen in any of our patients; as supposed by the author, it should be due to cortisone application. 3. As Jadassohnnaevus may develop basalioma or adenocarcinoma, further dermatologic survey is indicated. 4. So long, no neurologic symptoms have occurred. PMID- 9333664 TI - [Reference values in vision development of infants with clinical use of the Teller Acuity Cards]. AB - BACKGROUND: Using Teller Acuity Cards (TAC) for clinical visual testing, the question arose how our measurements fitted in the different standard tables of the producer's hand-book. In addition, we wanted to investigate how reliable the measurements of newborn and infants were and what the examination success rate under clinical conditions was. METHODS: At the paediatric clinic of the University of Erlangen, we tested the binocular grating acuity of 98 infants up to the age of one year, using the complete set of Teller acuity cards. In addition, 41 of the children underwent a monocular vision test. RESULTS: 1. Theoretical: At first we calculated conversion data for our card set. Using this conversion scale from cy/cm in cy/deg and the corresponding vision equivalent we produced our own standards for the development of grating acuity up to the age of one year. 2. Clinical: In 3-5 min per clinical examination we could determine for 90.8% of the patients a vision equivalent. The reliability of the results was age dependent and was at its best at the age of 5-11 months. The reliability was also very dependent on the duration of the test and the number of test runs. This resulted in a limited card choice for each age group. PMID- 9333666 TI - [Post-traumatic endophthalmitis with multiple water pathogens]. AB - HISTORY AND SIGNS: A 37-y-old worker for a recycling company suffered from a corneal perforation by a wire which had been in contact with dirt and moisture. THERAPY AND OUTCOME: Initially, the typical signs of an endophthalmitis were missing so that we only irrigated and instilled antibiotics into the anterior chamber. However, later on two pars-plana vitrectomies became necessary. At each operation a bacteriological examination was performed and we found four different species-Pseudomonas aeruginosa. Klebsiella oxytoca, Aeromonas caviae, Flavobacterium odoratum. CONCLUSION: The latter two species are very rare in ophthalmology. This fact and some peculiarities in the clinical course of the patient's disease make him an unusual case. After unsuccessful irrigation of the anterior chamber vitrectomy ought to be performed immediately. PMID- 9333665 TI - [Acute glaucoma caused by massive subretinal hemorrhage in age-related macular degeneration and disordered thrombocyte function]. AB - BACKGROUND: Hemorrhages from subretinal neovascularizations in age-related macular degeneration are not uncommon, but usually limited to the posterior pole. A therapy-resistant angle closure glaucoma following displacement of the lens iris diaphragm due to massive choroidal and subretinal hemorrhages has rarely been reported. CASE REPORT: A 72-year-old female patient was first seen in September 1993 because of decreased vision in the right eye. An age related macular degeneration with an extended untreatable neovascularisation membrane was diagnosed. One year later sudden pain attacks were initiated by secondary angle closure glaucoma, caused by a massive choroidal and subretinal bleeding. At first the IOP could be regulated by drug therapy, however, recurring hemorrhages led to continuously elevated IOP. The reason for the recurrent massive hemorrhages was found to be a disorder of the platelet function. Since the IOP (50-80 mm Hg) could not be controlled any further, we decided to perform a sclerotomy in an attempt to drain the blood. Within one day another bleeding occurred and again led to an increase in IOP. Finally, the patient agreed to have the eye enucleated. CONCLUSION: In case of massive hemorrhages in age related macular degeneration a haematological systemic disorder must be included in the diagnostic considerations. If the intraocular pressure can not be lowered neither medically nor surgically, enucleation can not be avoided. PMID- 9333668 TI - [Idiopathic conjunctival lymphangiectasia]. AB - PATIENT: A 41-year-old male was evaluated for multiple yellowish cysts on the temporal bulbar conjunctiva of his right eye. These cysts have spontaneously developed over a period of one week, have been punctuated, and recurred one week later in an even greater number. The patient's ophthalmologic and medical history were normal. Complete excision of the largest cysts and marsupialization of the smaller cysts were performed. Histologic features of the excised cysts were consistent with lymphangiectasia and displayed dilated lymphatic channels. CONCLUSION: Conjunctival lymphangiectasia is a benign condition of unknown etiology. It has been observed with many underlying diseases as well as spontaneously and the dilated lymphatic channels can secondarily become hemorrhagic (Hemorrhagic lymphangiectasia of the conjunctiva). Simple excision and/or marsupialization are therapeutic options. PMID- 9333667 TI - [Dural carotid-cavernous sinus fistulas: clinical aspects, diagnosis and therapeutic intervention]. AB - BACKGROUND: Carotid cavernous fistulas are cerebral artenovenous shunts which may present with ocular or orbital signs. Direct fistulas are distinguished from dural shunts with respect to the anatomical situation. PATIENTS: We report on two patients with spontaneous dural carotid cavernous fistulas with multiple feeding vessels. Both patients required endovascular embolization. RESULTS: Therapy was successful in both patients. We present an overview of the clinical picture, the diagnostic procedure, the differential diagnosis and the therapeutic possibilities in this clinical entity. CONCLUSION: Both patients prove the importance of an immediate differential diagnostical classification so that a specific neuroradiological diagnostic can be ensured. Today's advanced endovascular technology offers therapeutic options with less risk for the patient than in recent years. PMID- 9333669 TI - [Chloroquine-induced bull's eye maculopathy without electrophysiologic changes]. AB - BACKGROUND: Electrophysiologic findings are usually pathologic in patients with chloroquine-induced bull's-eye maculopathy. To avoid maculopathy the daily dosage of chloroquine is estimated not from the actual but from the ideal body weight and should not exceed 3.5 mg/kg/day. PATIENT AND METHODS: A 59-year-old housewife took a daily dosage of 250 mg chloroquine for her rheumatoid arthritis over a period of 5 years up to a total dose of 450 g. With the height of 160 cm she weighed 68 kg. In 1990, two years after cessation of treatment she complained about blurred vision. Her visual acuity then was 0.8 and fell to 0.3 (right eye) and 0.4 (left eye) in 1996. No vortex keratopathy was observed. A central scotoma was present and fundus-examination showed a typical bull's-eye maculopathy. The mid hypopigmented ring correlated with an increased background fluorescence in the fluorescence-angiogram. Color vision and the retinal nerve fiber photo were normal. In spite of the prominent fundoscopic changes the electrophysiologic examination of this patient (ERG, EOG and pattern-ERG) was normal. The relative smallness of affected retina might explain the normal electrophysiology. CONCLUSION: This case of a patient with typical chloroquine-induced bull's-eye maculopathy with normal electrophysiology points to the importance of ophthalmoscopic and visual fields examination in patients under long-term chloroquine treatment. The correct daily dosage of chloroquine below 3.5 mg/kg/day should be given to avoid maculopathy. PMID- 9333670 TI - [Endophthalmitis after intra-oral block of the infraorbital nerve]. AB - BACKGROUND: Most penetrating needle puncture injuries occur in retro- or peribulbar anesthesia. Hereby only a small percentage of patients develop endophthalmitis. Ocular penetration after enoral infraorbital nerve block has not yet been reported in literature. HISTORY AND FINDINGS: In June 1995 a 74-year-old man presented with a fulminant fibrinous and purulent endophthalmitis. Because he suffers from trigeminal neuralgia his anesthesiologist performed an infraorbital nerve block from enoral two days ago. During this procedure the patient felt a sharp ocular pain. THERAPY AND OUTCOME: We suspected an ocular penetration and performed a vitrectomy with intravitreal antibiotic instillation on the admission day. A needle penetration site near the inferior rectus muscle was detected and after exocryocoagulation a 5 mm wide radial buckle was sutured over penetration site. Three months postoperatively vision recovered from hand moving to 20/50 and all infiltrations had been disappeared. Only preexisting cataract prevented a better vision. 10 months later after successful cataract extraction with intraocular lens implantation patient left hospital with a vision of 20/30. CONCLUSION: Careful anamnesis would have prevented this accidental globe penetration. Right upper palate is absent presumably due to congenital cleft malformation or surgery. This allowed needle penetration through smooth tissue into the right globe. Fortunately, endophthalmitis develops only in a small percentage after needle puncture. We recommend immediate pars-plana-vitrectomy and intravitreal antibiotics in case of endophthalmitis after ocular penetration. PMID- 9333671 TI - [Idiopathic polypoid choroid vasculopathy]. AB - BACKGROUND: The idiopathic polypoidal choroidal vasculopathy, also known as "posterior uveal bleeding syndrome" or "multiple recurrent serosanguineous retinal pigment epithelial detachments in black women" is a rare disease entity. A clincopathologic correlation of a patient with this disease is presented. CASE REPORT: A 47-year-old black woman was evaluated for a decrease of visual acuity in her right eye which had occurred over the last 3 months. Ophthalmic examination of her right eye revealed sub-RPE hemorrhage associated with a reddish-orange subretinal vascular-like lesion. In addition, both eyes displayed a few choroidal vascular-like bulbous structures in the superior temporal peripapillary region. The patient developed an extensive choroidal hemorrhage that led eventually to the enucleation of the eye. CONCLUSION: Choroidal neovascular membranes (CNV) secondary to idiopathic polypoidal choroidal vasculopathy differs in many aspects from other entities associated with CNV including clinical and fluorescein angiographic features, clinical course, and prognosis. PMID- 9333672 TI - [Experience with a new method of assisted human reproduction based on statistical data from Germany for the year 1995]. AB - Androgynous subfertility is more and more often of unfertile marriages (40-50%). Through application of new method during assisted human reproduction, the method of in vitro fertilization (IVF) is upgraded by intracitoplasmatic spermatozoids initiation (ICSI). Very high percentage of fertilisation has been achieved with the extremely bad ejaculate with oligo-astheno-teratozoospenny (oat), as well as after microsurgical interventions on testies (mesa-tesa). Very high percentage of nidation (24%), with an average of 2.1 embryos per transfer surprises and encourages. PMID- 9333674 TI - [Free skin-adipose and mucous tissue autotransplantation in the reconstruction of atrophic changes in the orbit and scarring of the conjunctival sulcus in war and peace conditions]. AB - We presented two cases with volume deficit of orbit and conjunctival contracture of socket. Both of them were with superior sulcus deformity and conjunctival contracture and symblepharon. The patients underwent reconstructive plastic surgery with an implantation of autogenous dermis-fat graft in the superior sulcus and implantation of oral mucous in the cul-de sac in the same manner. Five months after the second operation we noted good cosmetic results. PMID- 9333673 TI - [Isolated war injuries of the thorax]. AB - From own clinical material the authors examined and analysed isolated injuries of thorax made by war injuries. The results are compared with the same from other authors in same conditions. Conclusion is that the fast evacuation to the first hospital urgent and if it is possible compensate the volume of blood, reanimation and drainage of pleural cavum. Considering the results the authors are making clear the indications for operative treatment for early and last thoracotomy and specially noticed contraindications for thoracotomy. The intensive care: is also very important and it has to be interdisciplinary, permanent and aggressive. The separate part is analysis of logistics in treatment of cases injury of thorax where the authors are showing their own experiences and making suggestions for solving the problem. PMID- 9333675 TI - [Our approach to the treatment of burns]. AB - The treatment of the burns is successful in situation when there is a separate department for burns with well-educated and highly skilled surgeons. The adequate reanimation during the first 48 hours is essential for the patients with burns to survive. Burns patients with circumferential burns of the body or the bower extremities should have at least three longitudinal incisions (escharotomy) prior to the development of the arterial ischaemia. The early tangential necrectomy should be performed between third and fifth day. The early grafting of the necrotic surfaces reduces the possibility of septicemic complications with deep burns and thus decreases the mortality rate. PMID- 9333676 TI - [On the 50th anniversary of the journal "Medicinski arhiv"]. PMID- 9333677 TI - [Percutaneous drainage of abdominal fluid collections guided by computer tomography]. AB - During the war, June 1992,-August 1994, at the Institute 20 percutaneous fluid collections and abscesses drainages in the belles were done, controlled by the CT. The percutaneous drainage we started in 1984, until now we had 141 cases, 20 cases in the war time, 14 of them were wounded, while the rest was suffering from a malignant process in stomach, pancreas, kidneys. Both groups had post-operative complications after liver injuries. 7 underwent the percutaneous drainage. The length was 1-64 days, drainage contents quantity was 60-5.000 m. The drainage was successful with 14 patients, while in 5 cases we had to repeat, change the catheter place. Only with 1 patients the drainage was not done, but an aspiration. The contents were send to microbiologic analysis. It was a retro peritoneal abscess collection. Based on our ten-years experience, we are of an opinion that the CT controlled percutaneous drainage is a very efficient, simple and acurata urgent radiology procedure. According to our experiences, nearly all cases were successful. PMID- 9333678 TI - [Surgical treatment of an anomaly of the conduction system in a case of supraventricular paroxysmal tachycardia with WPW syndrome resistant to four conventional antiarrhythmic agents]. AB - The authors have presented girl with supraventricular paroxismal tachyarrthymia and atipic WPW syndrome. The most frequently disrhythmias in childhood and at infants. Laboratories and others specific research are normal. SPT was proved with ECG. The Attacks SPT in girl are precipited usually nervous, fear, premenstruation phase or acute infection as in last hospitalization. In despite of many years medicament treatment attacks are repeated. The authors are presented diagnostics difficulties and advantages surgical intervention-ablation of A-V accesorius node (success 95%), after electrofiziologic investigations. PMID- 9333679 TI - [Pulmonary abscess treated with postural drainage]. AB - Case record of patients with lung abscess treated by postural drainage is presented in this paper. In young man with multiple explosive injuries lung abscess was formed two months after injury. A postural drainage with parenteral application of antibiotics has been performed. The expectoration was painful. At the seventh day there was no temperature, ESR was described at the tenth day. The general status was becoming better. At the seventeenth day patient was discharged from Hospital. Rig imaging was shown nearly completely resolution of lung abscess. Postural drainage was effective because of favorable localisation of abscess near the large bronchus and basely part of the lung. PMID- 9333680 TI - [Characteristics of knowledge and attitude to drugs and drug addiction in a population of addicted and non-addicted adolescents]. AB - Governed by the idea of importance of the extent and character of being informed on drugs and narcomany the author of the study were reviewing a pattern sample of 50 drug-addicts while patients of the Centre for Treatment of Drug-Addiction with the Psychiatric Clinic of the Clinic Centre in Sarajevo. The control group was composed of fifty pupils of secondary schools (also-from district of Sarajevo) who were positively determined of never having had any experience of drugs. The results of the study evidently distinguish between the experimental and the control groups, both in respect of sources and way of informing as well in certain viewpoints and opinions in its prevention etc. The author think that the very existence of obtained discrepancies in the said aspects speaks enough for itself, binding upon us to take them into account in all subsequent preventional activities and plannings. PMID- 9333681 TI - [Genetics of manic depressive disorder]. AB - Genetic transmission in manic depressive disorder (MDD) has been explored in linkage and association studies. The X-linked transmission hypothesis has been tested by using several markers on chromosome X. The hypothesis of autosomal transmission has been tested by association studies with the 0 blood group located on chromosome 9, as well as linkage studies on chromosome 6 and chromosome 11. Although linkage studies support the hypothesis of a major locus, several factors are limiting the results which are discussed in the present review. PMID- 9333684 TI - [Computerized presentation of densitometry values of the size of bone defects]. AB - On the basis of experimental knowledge, this investigation intended to give its contribution to knowledge of the process of bone reparation and regeneration of alveolar extension cyst's. The results have shown that the denzinometrics, as a possible way of reparational and regenerational processes verification, has its full X-ray and clinical justification as it enables not only the observation of the reparational process but also mutual comparison. PMID- 9333682 TI - [The effect of Kaviner and the smear layer on dentin permeability of permanent teeth in children]. AB - The dentin permeability is defined as a moving of fluid, of chemical substances and microbial products as well through the dentin. The clinical protection of children permanent teeth in other words the protection of pulp-dentin complex after their preparation makes a big problem into the restorative stomatology. Caviner is one of the newer means which is used in the protection of pulp-dentin complex which represents the components of powder dispersing within the ethyl acetate mixture of polystirol. The important variable in this study is the presence or absence of smearing layer which has the important influence onto the dentin permeability. In order to confirm the Caviner and smearing layer working onto the dentin permeability of the children permanent teeth in vitro experiment was designed. We used for it the dentin disks made of the intact first premolar, extracted because of orthodontic reasons at the ten years old children. They were put into the split chamber which represents a part of apparatus made at our Faculty, which is, in fact, a modified apparatus which was formed by prof. D.H. Pashley (Georgia, USA), and it is used for measuring of dentin permeability with the help of hydraulic conductance (Lp) of dentin. The dentin permeability is expressed by the hydraulic conductance term, but the measures are still expressed as the Lp percentage maximum because of better survey of results. Comparing the obtained results with the other authors results we have come to the similar conclusions, and that is in fact, that the smearing layer significantly reduces the dentin permeability, and that the Caviner, as the other layners, reduces the dentin permeability. If we compare the reduction of dentin permeability at the dentin covered with the smearing layer and with the Caviner, it has been noticed that the smearing layer reduces more significantly. It should be accented that the Caviner reduces the dentin permeability less than the most earlier researched means for the protection of pulp-dentin complex. PMID- 9333683 TI - [Unexplained deaths in war injuries in Bosnia and Herzegovina]. AB - The paper presents the research in cases of the unexpected and sudden of the patients injured by fire-arms, during the aggression on BH. On the basis of the results of the origin of injury, the period of providing medical aid, the level of tissue damage and the function disturbancy, it could be concluded that numerous factors which are typical for the state of war are included comarde to the influencing factor of injuries observed in the incidents in the times of peace. The injuries in the course of the aggression are characterized by a severe tissue damage which is frequently followed by the poison gases poisoning, the quality of the first aid given, delayed and aggravated transportation, multi organic disbalance of the morphological structures and functions as well as an increased development of these mediators which bring about this type of disbalance. The lack of chemical diagnostic means prevents the determination of existence of the indicators of threatening deaths in the cases where there are no clinical signs for a fatal outcome. The frequency of casualties has prevented the prompt control of the biochemics and thus resulted in the increase of the number of unexpected and unclarified deaths. PMID- 9333685 TI - [Use of health technology in the reconstruction of the health care system in Bosnia and Herzegovina]. AB - In this paper author gives a brief overview of current state of development process of health care system in B&h which was significantly changed during aggression to our country. Main part of medical facilities of that system was either destroyed, especially medical equipment, which is now unused old. An important segment in health care reforms in Federal program of health in B&H have to be gathering and using of modern medical chronologies based on microprocessors. In that case, we need start with education of medical staff in that field. PMID- 9333686 TI - [Development of gastroenterohepatology in Bosnia and Herzegovina]. AB - In this paper author described establishing and development of gastroenterohepathology, as independent disciplines, in Bosnia and Herzegovina. Also, he described development of Association for gastroenterology and Clinic for gastroenterohepathology of Clinical centre University of Sarajevo. PMID- 9333687 TI - [Analysis of intensity of post-stroke depression and its relationship with the cerebral lesion location]. AB - BACKGROUND: Several studies have suggested a close relationship between intensity of poststroke depression and lesion location, but the relationships are not clear for some authors. In this study we try to test the hypothesis that the intensity of depression in an early stage (4th week) of cerebral vascular accidents (CVA) in related to lesion location and type of lesion. PATIENTS AND METHODS: In a sample of one hundred and thirty patients in the fourth week of the evolution of an unilateral CVA, 48 patients fulfilled research diagnostic criteria (RDC) of post-stroke depression (34 major depression; 14 minor depression). Schedule for Affective Disorders and Schizophrenia (SADS) and standard measures of intensity of depression (Hamilton's HRDS, Montgomery-Asberg's MADRS and Beck's BDI) were used. RESULTS: In the fourth week of the evolution of CVA's, depression intensity measures (HRDS and BDI) were significantly higher in anterior lesion when compared to posterior lesions. No significant differences were found between right and left lesions, but both were higher than in non-hemispheric lesions. No significant differences of prevalence or intensity of depression were found in relation to lesion type (hemorrhagic/ischaemic). CONCLUSION: The results confirm a relationship between intensity mood disorders after a cerebral vascular accident and the topography of the lesion. PMID- 9333688 TI - [Assessment of the transcultural equivalence of the Spanish version of the profile of quality of life for chronic patients (PECVEC)]. AB - BACKGROUND: In this work the transcultural adaptation and validation of a quality of life questionnaire has been approached, taking into account its increasing interest and its practice considerations. The Profil der Lebensqualitat Chronischkranker (PLC) is a German instrument and titled in Spanish. Perfil de Calidad de Vida para Enfermos Cronicos (PECVEC). The main goal of the present study is the assessment of the transcultural equivalence between the Spanish version and the original questionnaire version. SUBJECTS AND METHODS: Translations and back-translations from the original instrument, using bilingual personnel, were made. The most important goal of the study was to achieve the more accurate equivalence between the original version and the Spanish version of the questionnaire. The translations were discussed between the groups of investigators in Germany and Spain and finally adapted a pretest that showed good statistical results. The instrument was applied in two groups of subjects paired by sex and age and its validation was made. RESULTS: The results of two studies (one German and other Spanish) are compared. In the Spanish study the Cronbach's Alpha coefficient (range: 0.39-0.97) and the factorial analysis of the items showed good internal consistency. Cronbach's Alpha was higher than 0.6 for all dimensions except for the scale of Social Well-Being (0.39) with an average value 0.72. The comparison between two factorial structures showed that both are similar statistically. The adapted version showed discriminant capacity between groups ("known group validity") and the final scores of quality of life were similar to the original version. CONCLUSIONS: The adaptation process from the original version to the Spanish version has finished with a transcultural equivalence between both. This preliminary validation can be seen as an important starting point to increase, in this field, the number of instruments of assessment health-related quality of life in chronic patients in Spanish language. PMID- 9333689 TI - [Fluctuating molecular remission in patients with chronic myeloid leukemia with long term complete cytogenetic response after alpha interferon treatment]. AB - BACKGROUND: Alpha interferon can induce complete cytogenetic responses (CGR) in nearly 25% of chronic myeloid leukemia (CML) patients, but few of them obtain molecular responses (i.e. disappearance of the chimeric RNA bcr-abl). The incidence and evolution of this response are not well known. PATIENTS AND METHODS: The molecular response has been explored in six CML patients, four in first chronic phase and two in relapse after bone marrow transplantation, who have obtained durable and sustained CGR. Bone marrow samples have been analyzed by double step RT-PCR. Results have been correlated with clinical and karyotype findings. RESULTS: Twenty nine samples have been studied. Every patient in first chronic phase CML have obtained molecular response, but this response has not been persistent. One of the patients with relapsing CML after showed transient PCR negativity, whereas persistent PCR positivity was detected in the other one. CONCLUSIONS: Interferon alpha can induce complete molecular responses in CML patients, but in this study this responses have been fluctuant in all the patients. PMID- 9333691 TI - [Streptococcus agalactiae (group B streptococci)]. PMID- 9333690 TI - [Stroke and depression]. PMID- 9333692 TI - [Nosocomial infection with methicillin resistant Staphylococcus aureus in 14 human immunodeficiency virus infected patients]. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) frequently occurs with nosocomial infections. Although human immunodeficiency virus (HIV) infected subjects spend a long time in hospital, the transmission of MRSA nosocomial infections in this group of patients has not been previously reported previously. PATIENTS AND METHODS: A clinical sample of 14 HIV infected patients from an Infectious Diseases Unit, in whom MRSA had been isolated, were evaluated as part of a 6 months prospective study. The measures employed in assessing infectivity were the prospective surveillance of all those isolated and the search for carriers in associated health-workers. RESULTS: The potential index case was a patient with an isolated MRSA pneumonia. From him, it was transmitted to his cohabitors and to the rest of the Unit. All the patients had AIDS and 13 presented with MRSA-associated symptoms. Five were admitted 30 days previously and 12 had intravenous catheters. The mean time for the appearance of the infection was 16 days. In the antibiotic investigations multiresistance was confirmed and in the 13 symptomatic cases systemic treatment with vancomycin was indicated requiring replacement by teicoplanin in 50% due to adverse reactions. Two years later, the 14 patients had died but only one in relation to MRSA. Of the health-workers, one carrier was detected. The line of decolonization with mupirocin was efficatious. CONCLUSIONS: The MRSA nosocomial infection in HIV infected patients took place in subjects who were immunodepressed and had a prolonged mean time of hospitalization. The treatment with vancomycin and/or teicoplanin was effective in the majority of the cases. PMID- 9333694 TI - [Nutrition and osteoporosis: a consolidated relationship]. PMID- 9333693 TI - [Antibiotic use and bacterial resistance to antibiotics: "something that concerns you"]. PMID- 9333695 TI - [Anaplastic variant of thyroid medullar carcinoma]. PMID- 9333696 TI - [Unexpected cholesterol measurement in patients with hypercholesterolemia]. PMID- 9333697 TI - [myotoxicity by chloroquine: report of a case]. PMID- 9333699 TI - [The arrival of the staff members]. PMID- 9333698 TI - [Phlegmonous gastritis in a patient with chronic gastric ulcer]. PMID- 9333700 TI - [1996 pathogen incidence and resistance status in peritonitis]. AB - Severe intra-abdominal infection is associated with a high mortality rate. In addition to risk factors in the patients, the causal pathogens and the selection of appropriate therapeutic procedures play an essential part in the course of these conditions. In the majority of intra-abdominal infections mixed aerobic/anaerobic infections, mostly with some involvement of enterobacteria and also of enterococci and staphylococci can be demonstrated. In addition to surgical intervention a calculated antimicrobial initial treatment of intra abdominal infections with an antibiotic with an adequate effect to combat the pathogen concerned can contribute to improving the patient's prognosis. A calculated antibiotic treatment can only be effectively and reliably carried through if the frequency of the pathogen and the resistance situation are known. Retrospective evaluations of data on the sensitivity and frequency of pathogens from a defined group of subjects allow conclusions on the epidemiological situation in a particular catchment area or in a medical sector and thus make it possible to calculate the appropriate therapy for infections. In 1996 a total of 2,779 bacterial isolates from the intra-abdominal infection sector were examined: 935 Enterobacteriaceae, 83 nonfermenters, 177 Staphylococcus spp., 211 Enterococcus spp., 39 Streptococcus spp., and 1334 different anaerobic bacteria. Fresh clinical isolates were available for all pathogens tested. The most frequent gram-negative pathogen was E. coli (60%) and the most frequent gram positive pathogen, E. faecalis (44%); the most frequent anaerobic pathogen was B. fragilis (39%). Taurolodine had the lowest resistance rate against gram-negative and anaerobic pathogens. Teicoplanin had the highest activity against gram positive pathogens. PMID- 9333701 TI - [Therapy of peritonitis today. Surgical management and adjuvant therapy strategies]. AB - Acute necrotizing pancreatitis and fylecal or diffuse purulent peritonitis are the diseases primarily responsible for mortality due to surgical infections of the abdomen. The most recent figures indicate that a mortality rate of 50%-80% in this specialized treatment group is still a reality. Without doubt, surgical sanitation of the focus is the most important therapeutic measure. A generalized inflammation reaction has been regularly observed in nearly all patients within this disease category. Local surgical therapy has the greatest effect on prognosis. If the therapeutic goal is not reached with the first intervention, adjuvant surgical therapy is necessary. The different forms are continuous peritoneal lavage (CPL), open dorsoventral lavage, and relaparotomy or scheduled reoperation ("Etappenlavage"). Adjuvant medical treatments include TNF alpha and interleukin-1 synthesis inhibitors or antibodies. Unfortunately, clinical studies with these mediators have only been partly successful in the subgroups, so that a general clinical adjuvant treatment is not considered viable. The bacterial properties of taurolidine destroy the bacterial membrane and, at the same time, lead to cross-linking of the membrane components and functional proteins (LPS), so that a bactericidal effect and endotoxin reduction take place simultaneously. Both local and intravenous routes of administration can be used. PMID- 9333702 TI - [Programmed lavage as a basic principle in therapy of diffuse peritonitis]. AB - The therapeutic concept of programmed lavage has been established in the treatment of severe diffuse peritonitis. This treatment was given to 30 patients from January 1995 to February 1997. The total lethality rate was 30% (11/30 patients). Ten patients had organ failure of three or four organ systems at the time of first laparotomy. In this group the lethality rate was 50%. On average 9.2 programmed relaparotomys were performed per patient. As early complications, small bowel fistulas were observed in three patients, bleeding in three patients and problems due to the laparostomy in four patients. PMID- 9333704 TI - [Adjuvant therapy of peritonitis with taurolidine. Modulation of mediator liberation]. AB - Despite aggressive surgical treatment, prompt antibiotic therapy, and modern intensive care, up to one half of patients still die of diffuse peritonitis. There must be a distinction between infection as a microbiological phenomenon and sepsis as a complex, deleterious, inflammatory host response. Physiologic and metabolic changes during the latter process by taxonomically different organisms or different sources of infection are often clinically indistinguishable. Taurolidine, an amino acid derivate, seems to cover a variety of effects in peritonitis. As secondary peritonitis is associated with a significant cytokine release that is compartementalized in the peritoneal cavity, taurolidine is bactericidal, antiendotoxic, and antiadherent locally and, on the other hand, may modulate the systemic cytokine-mediated inflammatory response after being adsorbed systemically by the peritoneum. Current management of peritonitis can clear the peritoneal cavity of microorganisms and their products but patients continue to die of uncontrolled cytokine-induced systemic inflammation. In patients that undergo daily staged, planned relaparotomies they should not only be treated locally by taurolidine but also systemically by intravenous administration. The latter should, as a sort of sequential therapy, be continued, especially when the peritoneal cavity has been closed after a series of relaparotomies. PMID- 9333703 TI - [Stage-oriented antibiotic therapy of peritonitis. Prospective study]. AB - PATIENTS AND METHODS: In a prospective protocol 25 consecutive patients with diffuse peritonitis were treated in the Surgical Clinic of the RWT-University in Aachen, Germany, from January to December 1995. According to the "Mannheim Peritonitis Score" three different stages were treated with different surgical procedures and a selective antibiotic regimen. Group-A patients with prognostically favorable peritonitis (MPS 0-20) were treated with the so-called standard procedure, group-B patients (MPS: 21-29) with closed postoperative lavage. The antibiotic regimen was cefotaxime (2 x 2 g) and metronidazole (2 x 500 mg) for both group-A and group-B patients. Severe group-C cases (MPS > 29) were treated with the so-called Etappenlavage (multiple reexplorations and intra operative lavage) and received a combination of three antibiotics (2 x 2 g cefotaxime; 2 x 500 mg metronidazole and 2 x 200 mg ofloxacin). RESULTS: Eight patients belonged to group A, 10 to group B, and 7 to group C. The mortality was 0% (group A), 20% (group B), and 29% (group C), respectively. The actual overall mortality of the whole group was 16% (4/25). The statistically expected mortality was 36%, according to the APACHE-II-Score (P = 0.0982). PMID- 9333705 TI - [Peritoneal instillation of taurolidine and heparin for preventing intraperitoneal tumor growth and trocar metastases in laparoscopic operations in the rat model]. AB - BACKGROUND: Although port-site metastases occur after laparoscopic surgery, there is no generally accepted approach to prevent tumor implantation so far. METHODS: In order to prevent tumor metastases, the effect of taurolidine and heparin on the growth of colon adenocarcinoma DHD/K12/TRb was measured in vitro and in a rat model. After incubation of the cells with heparin, taurolidine or both substances, the cell kinetics were determined. In a second experiment, tumor cells were administered intraperitoneally in rats (n = 60) and pneumoperitoneum was established over 30 min. Rats were randomized into four groups (I: tumor cells; II: cells + heparin; III: cells + taurolidine; IV: cells + taurolidine + heparin). RESULTS: While tumor cell growth was not influenced by heparin in vitro, growth decreased significantly after incubation with taurolidine and taurolidine/heparin. In vivo, intraperitoneal tumor weight was lower in rats receiving heparin (298 +/- 155 mg) and taurolidine (149 +/- 247 mg) than in the control group (596 +/- 278 mg). When the two substance were combined, tumor growth was even less (21.5 +/- 36 mg). Trocar metastases were only lower in rats receiving taurolidine or the combination of taurolidine and heparin. CONCLUSION: In vivo, heparin inhibits intraperitoneal tumor growth only slightly, while taurolidine causes a significant decrease in tumor cell growth in vitro as well as intraperitoneal tumor growth and trocar metastases in vivo. PMID- 9333706 TI - [Local antiseptic and anti-endotoxin measures in intra-abdominal infections]. AB - With standardized operating strategies, a lethality rate of 10.2% was achieved following intra-abdominal administration of taurolidine in 352 cases of severe intra-abdominal infection. The extent and type of antibacterial therapy were determined on the basis of the clinical severity, the patient's age, and the original site of the infection. Local antisepsis includes tactical surgery and the use of locally and systemically acting taurolidine. Antibiotics were used for systemic antibacterial therapy. After laparoscopical clearance of the focus of infection (appendix, gall bladder) the operating time was significantly extended compared with that required for open surgery, while the postoperative complication rate and the length of stay in hospital were significantly reduced. PMID- 9333707 TI - [Surgical therapy of pleural empyema with tauroline]. AB - Empyema continues to be a significant problem in spite of improved surgical techniques and the use of new, more potent antimicrobial agents. This report describes our experience in the treatment of empyema at the Clemens Hospital in Munster, Germany, from 1990 to 1996. Basic to conservative treatment are closed drainage with intensive irrigation and instillation of Taurolin, a chemotherapeutic agent against bacterias, yeasts and mycetes. This treatment has been employed since 1990 and given 86 patients with just empyema or in combination with decortication. The superiority of this method to other methods of treatment is discussed on the basis of our results. PMID- 9333710 TI - [Arteriosclerosis--an infectious disease?]. PMID- 9333709 TI - [Comparison of local and systemic inflammation after laparotomy or laparoscopy in the rat sepsis model]. AB - BACKGROUND: Laparoscopic techniques are frequently used in patients with peritonitis or intra-abdominal inflammatory diseases although increased intraperitoneal pressure may cause sepsis by promoting bacteraemia and systemic inflammatory response. METHODS: This experimental study investigates the influence of laparotomy and laparoscopy on bacteraemia, tumour necrosis factor (TNF)-alpha and endotoxin plasma levels. Standardized foecal inoculum was injected intraperitoneally and rats underwent either laparotomy (n = 20), laparoscopy (n = 20), or no further manipulation in the control group (n = 20). RESULTS: One hour after intervention, bacteraemia was significantly higher in both the laparotomy or laparoscopy groups than in the control group (P = 0.01). Foecal inoculum caused a significant increase in TNF-alpha and endotoxin plasma levels 1 h after intervention with the significantly highest levels after laparotomy (P < 0.05). In addition, the mean number of intraperitoneal abscesses were also significantly higher (P < 0.05) after laparatomy (n = 10) than after laparoscopy (n = 8) or in the control group (n = 5). CONCLUSIONS: Laparotomy and laparoscopy increased the incidence of bacteraemia and systemic inflammation compared to control group. However, inflammatory response and intraperitoneal abscess formation were significantly higher in the laparotomy group than in the laparoscopy group. PMID- 9333711 TI - [Endothelial receptor antagonists]. PMID- 9333708 TI - [Importance of mycoses in intra-abdominal infections]. AB - Although there is a 20% yeast colonization in the gastrointestinal tract of the population, fungal infections appear only rarely in secondary peritonitis. The risk of severe mycosis increases after a major operation and when a patient is taking broad-spectrum antibiotics, is on total parenteral nutrition, is catheterized, and/or is immune-suppressed. In the past years the incidence of nosocomial fungal infections (usually Candida spp.) has risen significantly. Five percent of CAPD-related peritonitis is caused by fungi. In enteral anastomosis breakdown, invasive mycosis occurs more often, with an accompanying lethality of up to 80%. In severe pancreatitis, up to 5% of peripancreatic necrosis is infected with fungi. The clinical course of severe mycosis, like the septic syndrome, is associated with fungemia in up to 50% of cases. As most of the facultative pathogenic fungi are part of the physiological flora, it is difficult to interpret mycological cultures. In order to diagnose invasive fungal infections, histopathological techniques and serologic tests for antigens and antibodies are available. Three antifungal agents (amphotericin B, flucytosine, fluconazole) are available for intravenous administration. Amphotericin B is given at doses of up to 1 mg/kg per day, in liposomal galenism up to 3 mg/kg per day. Combining amphotericin B with flucytosine (150-200 mg/kg per day) a synergistic effect is reached. Fluconazole at a dosage of 200-800 mg per day represents an alternative with similar antifungal activity and lower side effects. PMID- 9333712 TI - [Aerosols. Administration systems for pulmonary administration of drugs]. PMID- 9333713 TI - Overexpression of core-binding factor alpha (CBF alpha) reverses cellular transformation by the CBF beta-smooth muscle myosin heavy chain chimeric oncoprotein. PMID- 9333714 TI - [What is cyclothymia?]. AB - The term "cyclothymia" is being used with different meanings. DSM-IV and ICD-10 define "cyclothymia" or "cyclothymic disorder" as a long lasting, subeffective disorder with frequent shifts between hypomanic and (sub)depressive states. In the tradition of Kurt Schneider cyclothymia was understood as a synonym for manic depressive illness exclusively, while different personality typologies speak of a "cyclothymic" typus. Historically, the term was first used by the German psychiatrist Ewald Hecker in 1877. The definitions of DSM-IV and ICD-10 seem to be satisfactory in respect to reliability, but the nosological position of "cyclothymic disorder" is unclear. We review results concerning clinical symptomatology, comorbidity, biological parameters, personality (including the question of creativity), psycho- and pharmacotherapy as well as clinical course, which leave many questions open. Nevertheless, results in family studies support the idea that at least a fraction of "cyclothymia" is a mild or subclinical form of bipolar disorders. Until further research, which is urgently needed, we suggest that the term "cyclothymia" should be only used according to the guidelines of DSM-IV and ICD-10. PMID- 9333715 TI - [The central dopaminergic system and depression]. AB - This review presents data supporting the involvement of the dopaminergic system in depressive illness. Neuroanatomical research, animal research and clinical studies have shown that a dysfunction of the mesolimbic dopaminergic system could be associated with depressive symptoms. Furthermore, the chronic administration of antidepressants in animals provokes a sensitisation and enhanced expression of dopaminergic receptors. Although the dopaminergic system has received little attention in biological research on depression in the last decades, current research on the dopaminergic system is about to change this situation PMID- 9333717 TI - [Are negative cognitions symptoms of depression or also an expression of personality traits?]. AB - This study examined the changes from hospital admission to discharge of different categories of cognitive distortions - automatic thoughts, self-concepts and dysfunctional attitudes - in 67 patients with an ICD-10 diagnosis of a depressive episode. Furthermore, the relationships were analyzed between cognitive distortions, degree of depression, and personality dimensions (isolation tendency, extraversion, rigidity, schizoidia, frustration tolerance and neuroticism). There was a significant correlation between automatic thoughts and negative self-concepts on the one hand, and severity of depression on the other upon admission. There was a significant reduction of automatic thoughts and the negativity of self-concepts from admission to discharge. No significant association was observed, however, between the severity of depression and dysfunctional attitudes. In addition, dysfunctional attitudes showed a significant decrease until discharge only in patients with a full remission of depressive symptoms. These results underline the fact that automatic thoughts and self-concepts are more state-dependent than dysfunctional attitudes. Moreover - even taking into account the degree of depression - there was a significant relationship between the personality dimension neuroticism and all cognitive scales upon admission. This indicates that a high degree of neuroticism may facilitate the emergence of negative cognitions during a depressive episode. PMID- 9333716 TI - [The tryptophan depletion test. Basic principles and clinical relevance]. AB - The application of a tryptophan-free amino acid mixture (tryptophan depletion test) induces a rapid and substantial lowering of both total and free plasma tryptophan. Consequently, the brain serotonin content and also cerebral serotonin function are decreased. This method provides a paradigm to study the role of serotonin in the pathobiology of depressive disorders and their treatment modalities. Untreated depressed patients show few behavioral effects during tryptophan depletion. In depressed patients during an antidepressant or light therapy-induced stable remission, a transient depressive relapse was induced by tryptophan depletion. Healthy subjects with a genetic risk for affective disorder show worsening of their condition induced by tryptophan depletion. These findings indicate the relevance of altered brain serotonin function in the pathophysiology of affective disorders and strengthen the importance of serotonin in the mechanism of action of antidepressants. Since recently published studies revealed some evidence that the serotonergic system is directly involved in the pathophysiology of various psychiatric syndromes besides depression, it seems to be reasonable to evaluate the validity of the tryptophan depletion test also in non-depressed patients. PMID- 9333719 TI - [Dependency outcome of alcohol-dependent patients after inpatient detoxification and motivation treatment]. AB - In addition to psychiatric outpatient treatment, the detoxification of alcoholics in psychiatric hospitals is an important link between medical detoxification and psychosocial inpatient treatment aiming at abstinence. Integrated models of inpatient detoxification and motivation therapy focus on improvement of the motivation of patients towards further treatment of their alcoholism and towards abstinence. In our study 529 alcoholics underwent such treatment lasting between 2 to 3 weeks. A total of 469 patients (89%) were followed up after 8 months; 242 alcoholics (52% of the follow-up sample; 46% of the whole sample) achieved the treatment goal and started further treatment, mainly as inpatients. Fifty-four of these patients relapsed before they started further alcoholism treatment, but achieved abstinence afterwards; 60 (25%) of these 242 patients relapsed after the specific alcoholism therapy; 227 alcoholics did not start further treatment after the initial detoxification and motivation therapy. In this subpopulation, 113 (50%) relapsed during the follow-up period. Although this is not a controlled study, it can be concluded that integrated inpatient detoxification and motivation therapy has positive effects on the treatment behavior of alcoholic patients and enhances the probability of further abstinence. PMID- 9333718 TI - [Suicide rates in a community psychiatric service system]. AB - Reduction of the elevated suicide rates in psychiatric patients remains an important aim of psychiatric treatment. There has been an ongoing discussion on the question of whether the increased tendency to transfer long-term psychiatric patients from inpatient care to community psychiatric care might go along with increased suicide rates. In our community care system we found an age- and sex adjusted suicide rate 40 times as high as in the general population in the period between 1973 and 1993. Within the first 4 years suicide rates were significantly elevated. All suicides had the diagnosis of chronic schizophrenia or schizoaffective psychosis. In the non-schizophrenic group no suicide occurred during treatment. Our study shows that in community psychiatric care suicide rates in schizophrenic patients are significantly increased whereas suicide rates is non-schizophrenic patients may be decreased. PMID- 9333720 TI - [20 years unsuccessful prevention of bipolar affective psychosis recurrence]. AB - A now 73-year-old female patient suffered from bipolar affective disorder for the first time at the age of 28. Despite treatment with all known phase-prophylactic agents (i.e., lithium, carbamazepine, valproate, and various antidepressants) she relapsed frequently. To render matters even more difficult, she easily developed side effects with most medications, particularly anticholinergic ones. In order to reach effective longterm prophylactic treatment, we tried several not yet established therapeutic regimens like clozapine, high-dose L-thyroxine, and maintenance ECT. In this case report we discuss the available literature with special respect to clozapine and maintenance ECT. PMID- 9333721 TI - [Reversible neuropsychiatric side effects of lithium with normal serum levels. A case report]. AB - Lithium therapy in patients with manic depressive disorder occasionally causes neuropsychiatric side effects even at therapeutic serum levels. Those lithium induced symptoms can be polymorphous and may be hard to differentiate from other disorders and from other drugs' side effects. We report the case of a 56-year-old woman with manic-depressive illness, who developed neuropsychiatric side effects from lithium prophylaxis at therapeutic serum levels. The symptoms included disorientation, aphasia, ideatoric apraxia, parkinsonism, restlessness and severe sleep disorder. In contrast to previous reports, the casual role lithium was underscored by symptom increase after lithium re-exposure at therapeutic serum levels. The patient recovered completely after cessation of the lithium prophylaxis. PMID- 9333722 TI - [Mianserin-induced hypertension 2 weeks after discontinuation of tranylcypromine]. AB - As compared to other tri- and tetracyclics, antidepressant therapy in patients suffering also from cardiac disease with mianserin is known to be relatively safe. Hypertensive effects of mianserin have not been described so far. We report a case of 64 years old patient with a dilatative cardiomyopathy and left anterior hemiblock, who developed hypertension (maximum: 180/125 mmHg) during mianserin treatment. Previous treatment with tranylcypromine has been stopped two weeks before. After discontinuance of mianserin blood pressure rapidly came back to normal values. The possible noradrenergic and serotonergic mechanisms of this phenomenon are discussed in relation to pretreatment with tranylcypromine. PMID- 9333723 TI - [Pro-psychotic change of binocular depth inversion by sleep deprivation]. AB - Binocular depth inversion represents an illusion of visual perception. Such inversion does not occur in all cases, especially when objects with a higher degree of familiarity (e.g. photographs of faces) are displayed. Cognitive factors are assumed to override the binocular disparity cues of stereopsis. We tested the hypothesis that during sleep deprivation the human CNS is unable to correct the implausible perceptual information. Measurements of binocular depth inversion in perception of 3D objects were taken in sleep-deprived medical staff and healthy volunteers. The binocular depth inversion scores were highly elevated in the sleep-deprived group in comparison to the healthy volunteers. The data demonstrate a strong impairment of binocular depth inversion after sleep deprivation and support the view that sleep deprivation may be accompanied by a disorganisation of the interaction between sensory input and generation of perceptual hypotheses. PMID- 9333724 TI - [Georg Buchner, ICD-10 and the physician's basic attitude]. AB - The novel Lenz (1835) by the German author, social revolutionist and physician Georg Buchner (1813-1837) can be regarded as a prominent document in the history of medicine. On the one hand, Buchner's description of a schizophrenic disorder meets recent criteria; on the other, he hints at the physician's modern general attitude toward the patient. The latter is derived from his philosophy of art and political motives. PMID- 9333725 TI - [Neuroleptic malignant syndrome--sequela of an individually relatively high neuroleptic dosage?]. PMID- 9333726 TI - [Psychotherapy of depressive disorders: on the theoretical background and its clinical relevance]. PMID- 9333727 TI - Proteinuria and Progressive Renal Disease. Proceedings of the 3rd International Symposium. Amsterdam, The Netherlands, June 1996. PMID- 9333728 TI - Peer review and the New York State Dental Journal. PMID- 9333729 TI - [Surgical management of pulmonary aspergilloma]. AB - Between 1983 and 1996., 79 patients'--operated on for pulmonary aspergilloma- clinical data has been analysed. The patients were comprised of 67 males and 12 females, with a mean age of 49 years (range, 24 to 69). Previous lung disorders were observed in about half of the cases (most frequently tuberculosis), while in the other half aspergilloma was developed on the basis of (sub)-acute infections. The most common symptom was haemoptysis (in 45% of cases). Aspergilloma was diagnosed preoperatively (especially by typical chest x-ray) in 62 patients. In the other cases tb, lung cancer, pyosclerosis were suspected. 67 patients underwent pulmonary resection (50 lobectomies, 12 wedge resections, 5 pneumonectomies), 12 cavities were opened by cavernostomy. The postoperative mortality rate was 10.1%. The most frequent complications were bleeding, prolonged air leak, pleural rest space, empyema, bronchial fistula and wound infection, which were occurred in cases with bigger cavities near chest wall. In most cases with pulmonary aspergilloma surgery remains the only effective treatment. Operation has a lower risk in asymptomatic patients, without chest wall involvement. In several cases cavernostomy might be applied successfully. PMID- 9333730 TI - [Endoscopic ligation of bleeding esophageal varices]. AB - Endoscopic variceal ligation (EVL) is a new method for treating oesophageal varices, which was developed to reduce the high complication rate seen with endoscopic sclerotherapy. This technique consists of mechanical occlusion and thrombosis applying small elastic bands around the variceal channels in the distal oesophagus. We report 39 patients treated with EVL during the period from April 1995 to December 1996. The average age of patients was 50.1 years, 33 men, 6 women. Portal hypertension was caused by alcoholic liver disease in 36 patients. Seven patients died within 1 month. Varices were eradicated in 28 patients. Variceal eradication required an average of 3, 4 sessions. Active bleeding was controlled by EVL in 11 patients out of 12. Recurrent bleeding occurred in 14 out of 39 patients (35%). One oesophageal perforation occurred in connection with overtube placement. There were no other major complications. In conclusion EVL is an effective treatment for bleeding oesophageal varices. PMID- 9333731 TI - [Determination of diminished bone mineral density in ankylosing spondylitis]. AB - Bone mineral density was determined by DEXA method on the lumbal spine and on the femoral neck, on 44 patients suffering from ankylosing spondylitis. It was established that the osteopenie (low bone mineral density) was covered by the syndesmophytes. The comparative measurement showed a lower bone mineral density for patients, who don't have syndesmophytes on the L II-IV. vertebraes. PMID- 9333732 TI - [Rabid hedgehog in inner-city area of Budapest]. AB - A family found a hedgehog in the inner-city area of Budapest (1st District, Castle Quarter). While playing with it, the four members of the family got beslavered by the hedgehog who then died. The dead animal was subjected to obligatory veterinarian examination abduction, and it proved to be rabid. All members of the family had to be inoculated against rabies as the probability of infection could not be excluded by the epidemiological examination. Surveying the available literature, if could not be found data relating to hedgehogs' rabies. PMID- 9333733 TI - [Cerebral embolism caused by a right ventricular pacemaker electrode perforated into the left atrium]. AB - The authors present a new, unusual side effect due to retained pacemaker lead. The free tip of the lead caused cerebral embolism by perforating the interatrial septum. Transoesophageal echocardiographic study showed the lead crossing the interatrial septum. The total abnormal course of the lead was demonstrated by lateral X-ray picture. Diagnostic and therapeutic approaches regarding retained leads in malposition are also discussed. PMID- 9333734 TI - [Second "actor" in the Semmelweis drama: J. G. Riedel (1893-1870)]. PMID- 9333735 TI - [Franz Schubert (1797-1828), his illness and death]. PMID- 9333737 TI - [Autopsy]. PMID- 9333736 TI - [Melancholia is not only a medical term]. PMID- 9333738 TI - [Chronic, non-radicular lumbalgia]. PMID- 9333739 TI - [Biomechanics of lumbar instability]. AB - Several authors have tried to define segmental lumbar instability. Their definitions: increased antero-posterior translation, pathologic coupled motion, increased neutral zone, pathologic instantaneous center of rotation describe some mechanic findings occurring in the aging spine. However, there is no evidence that they help to differentiate the pathologic entity of segmental lumbar instability from the normal aging process. Dynamic explanation models are promising but at the moment they cannot be used clinically for diagnosis of instability as well. The most important structure to maintain lumbar stability is the intervertebral disc. In the third and fourth decade, more than 50 percent of specimen show peripheral tears of the anulus. It was shown in animal experiments that these tears develop to radial tears, which are accompanied by nuclear volume loss and decreased height. The facets degenerate one or two decades later. Corresponding with the loss of discal function, they increasingly contribute to spinal stability. In conclusion, the concept of lumbar segmental instability is not very helpful in clinical practise. It is recommended to base the decision of lumbar fusion on a painful degenerated disc, and additional findings promising a good result. PMID- 9333740 TI - [Psychosomatic aspects of chronic pain]. AB - This paper gives a report on the current views of psychosomatic theories on chronic pain. Based on Melzack's and Wall's "gate-control theory", Freud's ideas on hysteric neurosis, and Engel's "pain-prone patient" the unidirectional nociceptive model of pain is replaced by a cybernetic system of biopsycho-social etiology. The psychodynamic theories of pain-proneness, the narcissistic mechanism in the etiology of pain, the mechanism of conversion, states of psychovegetative tension and processes of learning are presented. In addition the impact of gain from illness, life-events, coping, the doctors behavior and the comorbidity of affective disorders on pain syndromes becoming chronic is pointed out. As an example the multifactorial etiology of chronic low back pain is demonstrated. PMID- 9333741 TI - [Is a preoperative evaluation program in treatment of patients with chronic lumbar syndrome useful?]. AB - The Pre-Operative Evaluation Program (PEP) in the USA is a 1-week outpatient program for patients who have physical pathology which could be improved with surgical intervention. Patients are prepared for surgery and their hospital stay through an education process that includes educational groups, understanding of anatomy, pain and stress management, psychological testing and exercises to improve strength and spine techniques to improve functional abilities. The orthopedic clinic in Kassel developed a 14-days inpatient preoperative evaluation and education program (PEP) for potential surgical candidates with chronic back or neck pain. PMID- 9333742 TI - [A psychological therapy methods in chronic non-radicular backache]. AB - The presence of psychological co-morbidity and difficulties in coping with chronic low-back pain are indications for the application of psychological therapy procedures. In the treatment of patients with chronic pain, the aim of therapy as well as the technique of the traditionally employed psychotherapeutic procedures need to be modified. What is of immediate importance is to promote an active pattern of coping with pain and discourage pain-related inactivity and reduce feelings of helplessness and hopelessness. To achieve this, it is necessary to combine various psychological treatment modalities (relaxation techniques and behavioural or psychodynamic therapy) with exercise programs as in physiotherapy or sports medicine. In the choice of the appropriate psychotherapeutic modalities, it is important to take into consideration the extent of chronicity of pain, the patient's subjective understanding of the illness, a possible wish of the patient for early retirement, previous experience with chronic pain and the extent to which the patient has been psychologically traumatised (especially in his childhood and youth). Relatives should be integrated in the planning and possibly also in carrying out of the treatment. Symptom-specific group therapy might motivate patients to undergo treatment and to change their habitual ways of life. A multimodal treatment approach incorporating orthopaedic, sports medicine, physiotherapeutic, psychological and socio-therapeutic procedures have proved to be more effective in the treatment of chronic pain than one employing monocausal treatment modalities. PMID- 9333743 TI - [Injection treatment of non-radicular lumbalgia]. AB - Low back pain is the most expensive condition in industrialized countries. Approximately 65-80% of the population will be afflicted with low back pain at some point during their life. Low back pain has many causes and can originate from any of several pain-sensitive foci, among which are facet joints, sacroiliac joint, muscle and ligaments. Primary care in the acute phase consists of nonsteroidal anti-inflammatory drugs to address the biochemical and inflammatory mediators of pain or skeletal muscle spasmolytics to reduce low back pain symptoms. Injection procedures should be reserved for the patients with low back pain who fail to respond to a directed, conservative treatment trial and have had pain for at least 2 weeks duration. Eliminating sensation from a certain pain source has been proposed as a way to allow an examiner to determine if that joint is responsible for the patient's pain. Injections of local anesthetic into the facet joint or around its nerve supply are clinical methods of eliminating pain from focal areas such as facet joints or myofascial trigger points. When a particular joint is determined to be the source of pain, long-term relief can be sought by directing therapeutic interventions at that joint. The anatomic accessibility of the most common pain sources of low back pain make diagnostic blocks and therapeutic instillation of corticosteroids particularly appealing. If used, their potential benefit for the individual case needs to be carefully weighed. They should be used to facilitate more aggressive conservative care and not as an isolated treatment. Certainly, if response to corticosteroids does not occur after the first injection, no further administration of corticosteroids is indicated. PMID- 9333744 TI - [Minimally invasive approach and surgical procedures in the lumbar spine]. AB - The history of minimally invasive lumbar spine surgery began in 1963 with the introduction of chemonucleolysis. Like this technique, the later development of mechanical nucleotomy and lasernucleotomy aimed primarily at reduction of the disc pressure. Miniature optical systems offered the opportunity for more specific decompression by discoscopy or, more recently, by transforaminal epiduroscopy. Initially, nucleotomy was the only feasible minimally invasive procedure. In recent years, however, minimally invasive spinal fusion became possible due to the development of new devices (Cages) and advanced transperitoneal (laparoscopic) and retroperitoneal approaches. PMID- 9333745 TI - [Advantages and disadvantages of retro- and transperitoneal approach for fusion of the presacral intervertebral disk]. AB - In a retrospective study, we evaluated 180 patients treated for painful spondylolisthesis with combined anterior and posterior fusion. Group I included 90 patients treated by anterior fusion with the transperitoneal approach. Group II included 90 patients operated on with a retroperitoneal approach. Group II showed a higher incidence of pseudarthrosis L5/S1 (4%), tear of the common iliac vein (1%), postsympathectomy syndrome (4%) and reversible L4 nerve-root lesion (3%). On the basis of our findings, we recommend the transperitoneal approach for anterior interbody fusion L5 to S1 or L4 to S1. PMID- 9333746 TI - [Lumbar fusion in adults--dorsal or combined ventral/dorsal approach?]. AB - A retrospective study was conducted to compare the results of posterior with combined anterior/posterior lumbar fusion in adults. Seventy-six consecutive posterior cases fused with pedicle screws and 46 combined cases were included and followed for at least 2 years. Subjective assessment was based on the Visual Analogue Scales, Waddel Disability and Impairment Score and the GBB for objective quantification of complaints. Furthermore, a thorough clinical examination was done and X-rays including flexion/ extension radiographs, were taken. Questions were asked about the occupational status as well. Pain decreased significantly more in the combined cases than in the posterior fusion cases. In one case a lesion of the common iliac vein occurred during a retroperitoneal approach. Only about half the patients working preoperatively returned to work again. In conclusion, the benefit of better pain relief after combined fusion must be regarded in relation to a higher complication rate due to a second approach. PMID- 9333747 TI - [Osteochondrosis dissecans]. PMID- 9333748 TI - [A new culicid in Italy: Aedes (Ochlerotatus) annulipes (Diptera, Culicidae)]. AB - The first italian record of Aedes annulipes is described. The species was collected in various sites of the eastern Po-Venetian valley (North-eastern Italy), from sea level up to 190 m a.s.l. The larval breeding sites were seasonal fresh water marshes within woods. Preimaginal development took place from February to May. Ae. annulipes was univoltine with a possible second minor generation. The females were strongly anthropophilic. Main morphological data are provided and compared with those of the close species Ae. cantans. The adaptation to environments south of Alps and even at sea level by a northern palearctic element such as Ae. annulipes is presumably achieved by the exploitation of sufficiently cold biotopes available for larval breeding during the winter-spring period. PMID- 9333749 TI - [Eco-epidemiological study of Phlebotomus perniciosus in foci of visceral leishmaniasis in Campania]. AB - An entomological survey was carried out during the sandfly season (June-October) of 1993 in three of the main visceral leishmaniasis (VL) foci of the Campania region of Italy. During the period 1989-1993, 39 VL cases were recorded in the three foci: 19 in Maddaloni, 14 in Ercolano and 6 in Ischia. For each focus, collecting stations representative of sandfly habitats (domestic, peri-domestic and sylvatic) located in urban, peri-urban and rural areas were selected for the sandfly investigation. Three monthly captures by means of sticky traps (20 x 20 cm) were carried out in each station. A total of 2,773 sandfly specimens were caught and identified as belonging to 4 species: Phlebotomus perniciosus (66.2%). P. mascittii (0.4%). P. papatasi (0.2%) and Sergentomyia minuta (33.2%). P. perniciosus, the proven vector of VL in Italy, showed the highest prevalence in all foci studied, Maddaloni (61.0%). Ercolano (62.5%) and Ischia (88.2%); the absolute density of the species (number of specimens/m2 of sticky trap) was not so high in all areas and habitats sampled, the values observed being between 0.14 and 22.05/m2. The relationship between the eco-epidemiological aspects and the distribution of P. perniciosus in the foci investigated is discussed. PMID- 9333750 TI - [Parasitic metazoans of Stenella coeruleoalba (Cetacea: Delphinidae) stranded along the coast of Latium, 1985-1991]. AB - The striped dolphin represents the most common species of cetacean stranded along the Italian coasts. A parasitological survey on 17 specimens of Stenella coerulecaiba stranded along coasts of Latium from 1985 to 1991, has been carried out. The morphological study enabled the identification of the following parasites. The sites are reported in brackets. DIGENEA: Campula rochebruni (liver), Campula palliata (liver), Pholeter gastrophilus (pyloric stomach). CESTODA: Tetrabothrium forsteri (intestine), Strobilocephalus triangularis (intestine), Monorygma grimaldii, larvae (abdominal cavity, mesentery, testes), Phyliobothrium delphini, larvae (subcutaneous fat). NEMATODA: Skrjabinalius sp. (lungs). COPEPODA: Pennella sp. (skin). ISOPODA: Ceratothoa parallela (mouth, stomach). AMPHIPODA: Syncyamus aequus (blowhole). PMID- 9333751 TI - [Observations of Setaria equina (Nematoda: Setariidae) with the optical microscope and scanning electron microscope]. AB - Adults of S. equina (Spirurida, Setariidae), 1 male and 2 females, collected from vaginal sac of stallion, were studied by soanning electron microscopy (SEM). The amphids, cephalic and cervical papillae, peribuccal ring, fine transverse bands and bosses of the cuticle, as well as caudal papillae were visualized clearly at this examination. The results of the present survey contribute towards the identification of S. equina, improve the definition of the characters which are demonstrated by common light microscopy and give the exact number of male caudal papillae including the lateral cloacal right papilla considered as a probable anomaly in literature. PMID- 9333752 TI - [Scanning electron microscopy study for an analytical key to the Hyalomminae (Ixodidae) species to be found in Italy]. AB - Identification of ticks of Hyalomminae subfamily is easy for the species of subgenus Hyalommasta Schulze (coxa l with two equal, stump and short, well separated spurs) and for those of the subgenera Hyalommina Schulze and Delpyella Santos Dias (basis capituli with lateral margins anteriorly divergent). All the species of subgenus Hyalomma Koch have the coxa I deeply divided in adjacent narrow external spur and wider internal spur, and the basis capituli rectangular, with parallel or subparallel lateral margins. The considerable intraspecific variation in the aforesaid subgenus practically reduces the diagnostic characters to the aspect of dorsal scuta and genital areas in the females and to those of same dorsal scuta in addition to adanals and subanals scuta in the males. In order to avoid further mistakes in the recognition of five species which are present in Italy too, viz. Hyalomma (hyalommasta) aegyptium, Hyalomma (Hyalomma) lusitanicum, H. (H.) detritum, H. (H.) excavatum and H. (H.) marginatum, some specific characters of adult forms were pointed out by scanning electronic microscopy (SEM) photos (for the males) and of semischematic figures (for the females), and utilized for a key to the above species. PMID- 9333753 TI - [Clinical and etiological aspects of goat ear mite infestations in Tuscany]. AB - Parasitic otitis associated with Psoroptes sp. mites was diagnosed for the first time in two flocks of goats located in Tuscany, Italy. Some animals presented clinical signs of ear mite infection, but the parasites were also isolated from the ears of two clinically silent goats. A morphometric study was conducted to establish whether there are significant morphological differences between mites collected from the ears of goats and Psoroptes cuniculi collected from the ears of rabbits. Three rabbits were experimentally infected with mites isolated from the auditory canal of goats, while nine goats were infected with mites isolated from rabbits. After three to five months all rabbits and five of the nine goats contained reproducing mite populations in their ears; also the morphometric analysis revealed no difference between rabbit and goat ear mites. It follows that Psoroptes cuniculi represents the etiologic agent of parasitic otitis in both these two animal species. PMID- 9333754 TI - [1st isolation of Aedes (Ochlerotatus) sticticus (Diptera, Culicidae) in the Po Venetian valley]. AB - Aedes sticticus is recorded for the first time in northern Italy. The species was collected in some relict woods mainly at sea level in the Po-Venetian valley (Northern-Italy). Larval growth took place in residual puddles originated by rain or river floods. The most abundant adult emergence was observed in April or May. Minor earlier or later adult emergencies were observed depending from egg submersion and temperature. Data on larval chetotaxy are presented. PMID- 9333755 TI - [Protozoan infections and intestinal helminthiasis among the population of a village in the northern Sudan savannah area of Mali (West Africa)]. AB - A cross sectional survey was carried out on intestinal parasites in a rural village located in a Sudan savannah area of Mali in September 1994. This survey was aimed to describe the prevalence of intestinal protozoa and helminths, and to evaluate the possible epidemiological impact of some sociobehavioural factors. A total of 209 stool specimens were examined with 3 methods: fresh stool examination, Kato thick smear technique and a formalin-ether concentration technique in a closed system. Cryptosporidium was also searched following the Ziehl-Neelsen staining and the immunofluorescence method using monoclonal antibody. Microsporidia were investigated by Trichrome staining technique. The concentration technique, as expected, was the more sensitive method: the protozoan cyst rate and the helminth egg rate were 70.3% and 11%, respectively. The low prevalence of intestinal nematodes, unexpected for this area, could be attributed to the improvement in sanitation (traditional WC present in 73.9% of the families), but also to the repeated treatments of the population against onchocercosis with ivermectin since 1992. We found no cases of isosporosis, cryptosporidiosis and microsporidiosis in our study population, neither significant association between socio-behavioural factors and parasitic infections. PMID- 9333756 TI - Fruit juice consumption. PMID- 9333757 TI - Fruit juice consumption. PMID- 9333758 TI - Fruit juice consumption. PMID- 9333759 TI - The case of the missing methylphenidate. PMID- 9333760 TI - The effect of tobacco smoke on children undergoing general anesthesia and surgery. PMID- 9333761 TI - Asthma compliance and psychological factors. PMID- 9333762 TI - Sedation for therapeutic and diagnostic procedures in children. PMID- 9333763 TI - Sedation for therapeutic and diagnostic procedures in children. PMID- 9333764 TI - Arrhythmias from Neonate to Adult. Symposium proceedings. The Netherlands, August 21-22, 1997. PMID- 9333765 TI - Pediatric emergency medicine: legal briefs. PMID- 9333766 TI - [Level of vasopressin in renal venous blood of patients with renovascular hypertension due to unilateral stenosis of renal arteries]. AB - Assessment of plasma renin activity (PRA) in renal vein blood is used in the diagnosis of unilateral renovascular hypertension (URVH). Recently also other markers of renal ischaemia (atrial natriuretic peptide, adrenalin, nonadrenaline and dopamine) have been described. The present study aimed to assess renal handling of vasopressin (AVP) by the ischaemic kidney in patients with URVH. In 16 patients with URVH, PRA and AVP were estimated in renal vein blood of the ischemic (IK) and non-ischemic kidney (NK), in arterial blood (A) and in blood samples withdrawn from the inferior vena cava (VCI) below the orifices of the renal veins. In contrast to PRA no significant difference between plasma levels of AVP in renal vein blood of the ischaemic and non-ischaemic kidney was noticed (4.8 +/- 0.9 pg/ml vs 5.1 +/- 0.8 pg/ml respectively). CONCLUSION: Chronic hypoperfusion of the kidney does not influence renal handling of AVP in patients with URVH. Thus assessment of AVP in renal vein blood in these patients is deprived of diagnostic value. PMID- 9333767 TI - [Nonspecific bronchial hyperreactivity in patients with seasonal bronchial asthma observed through two consecutive years]. AB - Seasonal bronchial asthma causally connected with the exposure to pollen allergens is a chronic, eosinophilic mucosal inflammation of airway. This inflammation is the basis for the development of nonspecific bronchial hyperreactivity which is the most typical but mutable feature of asthma. Bronchial hyperreactivity often determines asthma intensity and the need of asthma treatment. The nonspecific bronchial hyperreactivity over two consecutive years was evaluated in 11 patients (2 women and 9 men) with seasonal bronchial asthma, sensitized to grass, remaining under the conditions of natural allergen exposure and out of this period. Bronchial reactivity to histamine was measured by Cockcroft's at all method. So called histamine threshold (PC20H) in mg/ml was assessed. The values of ventilatory parameters (FVC, FEV1) and asthma symptom scores were also measured. It was showed that nonspecific bronchial hyperreactivity significantly increased in subjects with seasonal bronchial asthma during natural pollen exposure. PC20H in two studies performed during this period decreased 3 and 6 times when compared to preseasonal values. The majority part of patients (80%) has the increased bronchial reactivity to histamine also beyond the of grass season when clinical symptoms of asthma and rhinitis are not observed. This postseasonal hyperreactivity could be the effect of the chronic inflammation process persisted from the period of natural allergen exposure. Continuous subthreshold, which means asymptomatic exposure to perennial allergens to which most of patients are sensitized, could be another reason of this hyperreastivity. The possibility of exposure to the activity of seasonal allergens the whole year in persons with asthma can not omitted, as the presence of pollens in the sample of the house dust in patient's flat is observed during the yield of pollen season. Nonspecific bronchial hyperreactivity in individual patients is fluctuated, which probably is not dependent on the intensity of natural allergen exposure. PMID- 9333768 TI - [The effect of repeated use of cuprophane and polysulfone dialyzers during hemodialysis on the count of natural killer cells in blood]. AB - The aim of the study was to compare the effect of hemodialysis with the reused cuprophane and polysulfone dialyzers and bicarbonate dialysis on the count of natural killer cells in the peripheral blood in patients with chronic renal failure during the hour of haemodialysis. The study was performed in 16 patients with chronic renal failure just before haemodialysis (0') as well as 15 and 60 minutes after the beginning of haemodialysis with the first and the fourth use of membranes. The count of natural killer cells (CD3-, CD16+) in the peripheral blood was assessed using the flow cytometry. CONCLUSIONS: 1) The count of natural killer cells (CD3-, CD16+) decreased transiently in the peripheral blood in observed patients during haemodialysis. 2) The use of new cuprophane membrane decreased the count of natural killer cells in peripheral blood during haemodialysis significantly more than the haemodialysis with the use of polysulfone membranes and reused cuprophane membrane. 3) The count of the natural killer cells (CD3-, CD16+) in the peripheral blood in patients with chronic renal failure assessed 15 minutes after the start of haemodialysis could be a marker of dialysis membrane hemocompatibility. PMID- 9333769 TI - [Significance of atrial signal-averaged electrocardiogram analysis in diagnosis of paroxysmal atrial fibrillation in patients with mitral valve prolapse syndrome]. AB - The aim of this study was to assess the clinical usage of recording time-domain parameters of atrial signal-averaged electrocardiogram (ASAECG) in diagnosis of paroxysmal atrial fibrillation (PAF) of patients with mitral valve prolapse (MVP). 85 patients with MVP recognized by echocardiography were divided into two groups: group I (MVP-PAG/+/) 41 pts (15 male and 26 female) mean age 37.1 +/- 8.9 with previously electrocardiographically documented episode of PAF, group II (MVP PAF/-/) 44 pts (20 male, 24 female) mean age 39.1 +/- 14.3 without PAF. The control group III consisted of 35 persons: 24 male and 11 female in mean age 37.7 +/- 6.2 without any cardiovascular diseases. All patients underwent additional investigations included: T3, T4, electrocardiography, exercise-test with moving "running track", 24-hours monitoring ECG with Holter's method and ASAECG recording. The following time-domain parameters of ASAECG were calculated: the root mean square voltage of the terminal 10, 20, 30 ms of the filtered P-wave (RMS 10, 20, 30) and total time duration of filtered P-wave (PWD). The adaptation of time-domain parameters of atrial signal-average in differential diagnostics of PAF during MVP has appeared as useless from clinical point of view. PMID- 9333770 TI - [TSH-receptor antibodies in thyroid diseases]. AB - The clinical usefulness of detection of TSH receptor antibodies (TRAK-assay, Henning) in differential diagnosis of thyroid disorders and in follow up of treatment of Graves' disease was evaluated and compared to thyroperoxidase antibodies estimation (DYNOtest antiTPO Henning). 313 patients with various thyroid diseases and 50 persons from control group were examined. Thyroperoxidase antibody (TPO-Ab) titers were frequently elevated in Graves' disease before treatment (88.6%, median 3361 U/ml) and in Hashimoto thyroiditis (100%, median 6055 U/ml). Median TPO-Ab was in normal range in other thyroid disorders and in control group (toxic nodular goiter: 58.5 U/ml, neutral nodular goiter: 46.0 U/ml, thyroid cancer after thyroidectomy: 58.8 U/ml, control group: 0 U/ml). TSH receptor antibodies (TRAb) were detected in 94.1% in patients with Graves' disease (median 52 U/l) and only in 12.5% with Hashimoto disease (median 4.1 U/l), in 25% with toxic nodular goiter (median 4.1 U/l), in 10.9% with neutral nodular goiter (median 4.6 U/l) in 17.4% with thyroid cancer (median 1.6 U/l) and in 4.8% in control group (median 1.7 U/l). The TPO-Ab titers did not decrease significantly during thyrostatic treatment of Graves' disease. Patients in clinical remission after treatment exhibited TPO-Ab in 76.9% (median 615.5 U/ml). In contrast, the TRAb titers failed in patients treated with thyrostatics longer then six months (64%, median 16 U/l). After treatment 86.7% of them had normal values (median 1.0 U/l). The increase of TPO-Ab persisted in patients investigated two years after subtotal thyroidectomy (92.3%, median 750 U/ml). Only in patients operated before 10 years the titers declinedsignificantly (32%, median 43.4 U/ml). The TRAb titers fell significantly after thyroidectomy (two years: 62.5%, median 10.7% U/l, 10 years: 36.4%, median 4.2 U/l). Estimation of TRAb is a powerful tool for differential diagnosis of Graves' disease and enables better monitoring of the applied treatment than estimation of TPO-Ab. PMID- 9333772 TI - [Role of restricting dietary sodium in treatment and prevention of essential hypertension--state of knowledge in 1996]. PMID- 9333771 TI - [Rheumatoid arthritis as a risk factor for development of multiple myeloma]. AB - 7 out of 154 patients with multiple myeloma (MM) with concomitant rheumatoid arthritis (RA) (5 persons) and Bechterev disease (BD) (2 persons) have been presented. There were 5 women and 2 men at age from 52 to 67 years. Four of them had joint's disease for 4, 5, 24 and 25 years prior to MM, and in the next there MM was diagnosed simultaneously with RA. Two patients are still living (50 and 55 months from the diagnosis of MM), the mean survival time of the five already dead was 34.5 months, and did not differ from the survival of patients with MM alone. The contribution of interleukin-6 (Il-6) and adhesion molecules ICAM-1, VCAM-1, CD44 in pathogenesis of both diseases are discussed. PMID- 9333773 TI - [Minimal residual disease in acute leukemias--clinical significance]. PMID- 9333774 TI - [A patient with a prosthetic heart valve]. PMID- 9333775 TI - [Homocysteine as a risk factor for vascular changes]. PMID- 9333776 TI - [Systemic lupus erythematosus with renal involvement: progress in research on pathogenesis, diagnosis and treatment]. PMID- 9333777 TI - [Professor Nanna Svartz--life and achievements of the honorary member of the Polish Society of Internal Medicine]. PMID- 9333778 TI - [Report from the IX Scientific-Educational Conference of the Polish Society of Nephrology 10-12 October 1996 Wloclawek]. PMID- 9333779 TI - [Unending history of haptoglobin--the last five years]. PMID- 9333780 TI - [Antioncogenes--tumor suppression genes]. AB - Recent knowledge about biological role of tumor suppressor genes and their products: RB1, p53, WT1, DCC, APC/FAP, NF1, NF2, VHL, MCC and MTS1 is presented. The main approaches of these agents as physiological regulators of cell growth and proliferation are discussed. Views on the tumor suppressor genes involvement in the development of inherited and sporadic forms of cancer have been reviewed. PMID- 9333781 TI - [The role of free radicals in the etiopathogenesis of systemic sclerosis]. AB - This article reviews the current concept in the ethiology and pathogenesis of systemic sclerosis. It is suggested that free radicals play a crucial role in pathomechanisms of scleroderma. In addition, the influence of some environmental agents (silica, bleomycin, alcohol, toxic oil) on free radical production and subsequent induction of scleroderma or scleroderma-like syndrome is also described. PMID- 9333782 TI - [Erythrocyte pyruvate kinase--an enzyme that may have an influence on oxygen transport to tissues]. AB - Hemoglobin, the critical protein in the delivery of oxygen to mammalian tissues, is poorly adapted to that function. This awkward situation is remedied by the presence in the red cell of 5 to 7 mM 2.3 DPG, which binds to Hb competitively with oxygen and reduces oxygen affinity. How the levels of 2.3 DPG in the red cell are regulated is an important question that has not yet been fully answered. The best established correlation with 2.3 DPG concentration in red cells is the activity of the enzyme pyruvate kinase. Inverse relationship between 2.3 DPG content and pyruvate kinase activity is the result of two conditions within the erythrocyte. The metabolites between FBP and PEP are in a state of quasi equilibrium because the activity of pyruvate kinase is so much lower than the activities of other enzymes in the pathway. Furthermore, pyruvate kinase operates, in vivo at a PEP concentration well below the Km concentration. In consequence, an increase in the glycolytic rate or inhibition of pyruvate kinase causes an increase in PEP concentration. Increases in PEP levels lead to increases in the levels of 2.3 DPG, and hence to increase in the level of 2.3 DPG via the 2.3 DPG synthase reaction. This relationship is demonstrated by the frequent occurrence of elevated levels of 2.3 DPG in pyruvate kinase deficient erythrocytes and by the decreased levels of 2.3 DPG and PEP which are observed in erythrocytes containing a pyruvate kinase with abnormally high activity at low PEP levels. It is thus clear that control of pyruvate kinase activity is a means to the control of oxygen delivery by the erythrocyte. It remains to be discovered whether any of the observed variations in human PK activity are due to reversible posttranslational modification and whether the potential for control of oxygen delivery via changes in pyruvate kinase activity is made use of the normal human adult. The availability of human full length of cDNA for PK should accelerate our understanding of he structure-function relationships of PK deficiency and enhance the possibility of gene therapy for seriously affected PK patients. PMID- 9333783 TI - [Lipids in the cell nucleus]. AB - The paper contains available data on the content, composition and metabolism of different lipid fractions in the nuclei. The results on physiological function of the nuclear lipids are also included. Nuclear phosphatidylinositols have been shown to play a role of messengers signalling from the cytoplasm to the nuclei. Most sphingomyelin is located in the nucleoplasm and it effects stability of DNA. A role of nuclear triacylglycerols remains unknown. Polyunsaturated long chain fatty acids control transcription of genes encoding lipogenic and glycolytic enzymes. Cholesterol present in the nuclear envelope plays only structural function, but that present in chromatin may be involved in regulation of cholesterol biosynthesis. PMID- 9333784 TI - [Neurotensin--structure, origin and biological function]. AB - Neurotensin is a 13-amino acid hormonal peptide which was first isolated from bovine hypothalamus. It is present in the digestive tract as well as in the central nervous system. It has a variety of biological activities as a central neurotransmitter or neuromodulator, and a peripheral hormone. NT receptors have been characterized in a variety of tissues and cell lines of peripheral and central organs. The physiological functions of NT include stimulation of pancreatic and biliary secretion, stimulation of colonic motility, inhibition of small bowel and gastric motility, trophic effect on numerous tissues of the gastrointestinal tract. NT exerts hypothermic and analgesic effect when injected into the central nervous system. From a clinical standpoint, studies with NT have led to implications of its involvement in schizophrenia, Parkinson's disease and Alzheimer's disease. PMID- 9333785 TI - [Sidero-fibrosis of the lungs after decades of arc welding]. AB - The case of a patient is described who suffered from pulmonary siderofibrosis, histologically confirmed as a long-term cause of arc welding for several decades. In spite of this, there was no severe alteration of lung function. Pulmonary siderosis in welders was considered to be a benign pneumoconiosis. However, in recent years it has been noticed that siderosis is accompanied by disorders of pulmonary function, depending in particular on the quality of the working place, technology of welding, and duration of the exposition. Especially in smaller workshops without medical service and regular control of the craftsmen, unfavourable working conditions are frequent. PMID- 9333786 TI - [Placentoid malformation of the lung as differential diagnosis of localized emphysema]. AB - In a 50-year old patient with a long history of chronic obstructive airway disease and pulmonary emphysema, unusual solid spongious areas adjacent to bullous tissue were detected by bullectomia because of mediastinal displacement and dyspnoea on exertion. Pathological anatomical diagnosis showed villous framework in the marginal regions of bullous transformed parenchyma. According to pathognomonic histological finding the lesion is known as placentoid malformation or placentoid bullous transmogrification, respectively. This disease must be differentiated against rare cystic tumours such as alveolar adenoma or sclerosing haemangioma as well as congenital lesions e.g. adenomatoid cystic malformation. The lesion presented here includes hamartomatous features, such as the presence of leiomyomatoid proliferations of smooth muscle cells and fatty tissue embedded in the villous stroma. The clinically predominant emphysematous transformation of the adjacent lung tissue is pathogenetically the result of a valve formation in combination with unphysiological traction forces. The ectatic lymphatic vessels in peripheral tissue may perhaps be of etiological importance. According to former studies this may be an congenital malformation with progressive development. Resection of affected lung parenchyma seems to be curative: so far, no recurrences have been noticed. PMID- 9333787 TI - [A new portable monitor for long-term cough recording]. AB - Cough is a common symptom of pulmonary diseases. For a number of reasons it would be of interest to have information about the frequency of coughs over a given period of time. So far, the cough recorders which are available are either too expensive or unwieldy. Hence, we developed a cough recorder linked to a portable, commercially available actigraph (about the size of a pack of cigarettes) that records coughing as an acoustic signal and ventral thorax movement. The signals are filtered via a band pass and sampled by a peak detektor with different time constants to separate the impulse character of the cough signal from the background noise level. The cough recorder registers coughing cumulatively over a period of one minute and has a storage time of one week. Since the acoustic signals are essential for the interpretation of the recordings, the analogue circuit was subjected to a separate validation programme. For this purpose, the distinction between active coughs of 10 volunteers (total number of coughs 550) and background noises (male and female voices and other defined noises, total number of noises 336) was tested. The complete assembly was then tested over night on 7 hospitalised patients with chronic cough. An infrared video camera system was used to make a reference recording of the overnight coughing. The results show that nearly every cough of the 10 volunteers was recorded (r = 0.99). 97.1% of the background noise was correctly interpreted. The complete recorder assembly correctly recorded 98.9% of the coughs (total 870) in the 7 patients. 4.8% of the background noise was erroneously registered as coughing. Summing up, it can be said that the portable cough recorder affords accurate recording of coughing over a period of one week, correctly distinguishing coughing from background noise. PMID- 9333788 TI - [Bioavailability of a new theophylline chewable pill]. AB - Oral administration of theophylline as a chewable tablet is an alternative to the conventional parenteral route used in the treatment of acute dyspnoea in asthma bronchiale and other obstructive pulmonary diseases. To investigate the bioavailability of this preparation, a randomised 2-period cross-over study on the pharmacokinetics of a 100 mg theophylline single dose was conducted comparing oral administration as a chewable tablet (test medication) or as solution (reference) in 14 healthy male volunteers (age 21-31 years, body weight 60-90 kg). Bioequivalence of the tested formulations could be confirmed basing on the primary pharmacokinetic parameters (AUC, Cmax) for the extent and rate of theophylline absorption. The criterion of bioequivalence was also met for the secondary parameters, including terminal elimination half-life, mean residence time, time point of maximal plasma concentration and the ratio Cmax/AUC. Hence, the chewable theophylline tablet is bioequivalent to theophylline given as a solution. With regard to therapeutic efficacy, equivalence to medication by droplets can be expected. PMID- 9333789 TI - [German Society of Pneumology: Recommendations for quality assurance in ambulatory and inpatient pneumologic management. "Quality Assurance in Pneumology" Working Group]. AB - Steps to safeguard quality assurance have always been an inseparable part of any action taken by physicians who are aware of their responsibility towards their patients. Quality assurance can be improved further by introducing additional procedures, some of which were developed by the industry, to ultimately achieve a comprehensive quality management. Of course, additional quality assurance measures involve additional cost and effort, so that the increasingly straitened financial circumstances of the hospitals restricting outpatient and inpatient care services imply a critical assessment and discussion of the pros (efficiency) and cons (cost factors) of such services. The article discusses in particular such procedures that are particularly well suited for application in pneumology. The establishment of "standards" via guidelines and recommendations and their continual updating are basic prerequisites for quality assurance. Many guidelines and recommendations to this effect have been worked out by the pneumological associations. As far as internal quality management in hospitals is concerned, it appears meaningful to delegate quality assurance work to selected supervisory staff members in different spheres of activity (medical care, nursing, hotel services, organisation of workflow) who also keep quality recordes. To establish external quality control of inpatient pneumological care it would be feasible to establish "peer review" techniques assessing random samples of patient records and of the attending rounds, as well as of the results of general inspections of the hospital. Discipline-specific quality indicators and participation in oncological aftercare programmes are also important. In the outpatient sphere, discipline-specific quality circles offer extensive possibilities for effecting quality assurance measures. PMID- 9333790 TI - [Appetite depressant drugs and the risk of primary pulmonary hypertension]. PMID- 9333791 TI - [Prognostically relevant parameters in patients with coronary heart disease, arterial hypertension and sleep apnea disorders]. AB - Patients with untreated sleep apnea syndrome have a higher cardiovascular mortality. It is not known which mechanisms lead to this increase in mortality and whether it is independent from the often associated coronary heart disease and systemic hypertension. In 48 consecutive patients with coronary heart disease confirmed by angiography, exercise-ECG, Holter-ECG, echocardiography, spirometric tests, analysis of ventricular late potentials, heart rate variability and a test for sleep-disordered breathing with a screening device were performed. Seventeen patients showed disordered breathing during sleep (obstructive sleep apnea) with a desaturation index of > or = 10 (mean desaturation index 17.3 +/- 9.3 vs. 2.6 +/- 3.1 in the patients without sleep-disordered breathing). There are no significant differences in age (58.9 +/- 6.1 vs. 59.7 +/- 7.6 years), body-mass index (28.6 +/- 3.7 vs. 27.7 +/- 3.3 kg/m2), left ventricular ejection fraction (57.2 +/- 13.6 vs. 64.0 +/- 14.6%), forced expiratory volume in 1 second/vital capacity 95.4 +/- 13.9 vs. 92.9 +/- 11.2% predicted, heart rate variability (standard deviation of the RR-intervals 39.4 +/- 29.4 vs. 37.2 +/- 17.0 ms), the frequency of premature ventricular beats over 24 h and at night, the frequency of multivessel disease (71 vs. 68%), additional hypertension 53 vs. 48%), status postmyocardial infarction (47 vs. 48%) and positive late potential analysis (24 vs. 13%). There were no ST segment depressions during the night. Patients with coronary heart disease and mild sleep-disordered breathing show no significant differences in the investigated parameters compared with patients without obstructive sleep apnea or sleep-disordered breathing. PMID- 9333792 TI - [Nasal CPAP therapy of obstructive sleep apnea syndrome with expiratory pressure reduction: a prospective randomized study of acceptance of treatment during therapy initiation]. AB - It is often difficult to achieve adequate acceptance of nasal continuous positive airway pressure (CPAP) therapy by patients with OSA. Many patients find it particularly inconvenient to expire against the treatment pressure. With this in mind, we have attempted to improve acceptance of CPAP therapy by using a bilevel system that reduces the treatment pressure during expiration. 52 patients were randomized either to initial treatment with CPAP therapy followed by bilevel treatment, or to treatment in reversed order. During bilevel therapy the ratio of inspiratory to expiratory pressure was fixed at 1:0.6. After each treatment the patients were interviewed on the basis of visual analogue scales to establish their subjective evaluation of such parameters as general well-being, quality of sleep, comparison of the respective treatment pressures, and possible preference for one of the two systems for long-term treatment. The minimal effective inspiratory treatment pressure during bilevel therapy (IPAP) and the minimal effective CPAP pressure were closely correlated (r = 0.89). There was no difference between the apnoea hypopnoea indices during CPAP therapy as compared with bilevel therapy. Most patients (57%) felt the treatment pressure with the bilevel system to be lower (p = 0.048). There were no differences in patients' well-being early in the morning, or in their assessment of sleep quality. The majority of patients (52%) preferred bilevel therapy for long term treatment, while 38% preferred CPAP therapy (n.s.). In a subgroup of 13 patients with a treatment pressure of > or = 10 mbar during CPAP therapy 10 patients (77%) gave preference to the bilevel system (n.s.). In a considerable number of patients the acceptance of treatment can be improved by using a bilevel system for initiation of nasal positive pressure therapy. PMID- 9333793 TI - [Automated CPAP--use for obstructive sleep apnea syndrome]. PMID- 9333794 TI - [Unattended continuous positive airway pressure titration--clinical relevance and cardiorespiratory risks]. PMID- 9333795 TI - [Effect on the duration of mechanical ventilation of identifying patients capable of breathing spontaneously]. PMID- 9333796 TI - [Gustav Killian Memorial Lecture. 100 years bronchoscopy--early history and subsequent development]. PMID- 9333797 TI - [Endobronchial ultrasound]. PMID- 9333798 TI - [Etiology and reproducibility of blood gas findings during stress testing in patients with obstructive respiratory tract diseases]. AB - After the stabilization of the obstructive airways disease 14 patients performed 4 times exercise-tests of 6 minutes at a bicycle during one week. The basic values of the lung function showed with FEV1 of 66%, Rt of 5,74 hPa/l/s and IGV of 155% a strong disturbance of the mechanical features of the lung. At the mean during the exercise tests the PO2a remained with 69 Torr constant. PC2a increases at the same time for 3 Torr. After the exercise test a significant increase of PO2a of 11.4 Torr takes place. At the individual patients there was a great variability of the results between the different tests. Only three patients showed always a decrease of PO2a-values. The reason for this variability is the ventilation/perfusion inhomogeneity, which changes between the investigations. There are significant multilinear correlations between the FEV1%- as the IGV% values and PO2a for the values at rest, during exercise as 5 min after the end of exercise. The exercise tests show at the PO(2a)-values the variability of the ventilation/perfusion inhomogeneity which dominates the bloodgas values. The important increase of the PO2a after the exercise test even at this patients can be of importance. The duration of this effect after multiple loads and the dependence of the workload should be investigated. PMID- 9333800 TI - [Tumor-associated prognostic parameters in non-small-cell bronchial carcinoma]. PMID- 9333799 TI - [Stress test blood gas analysis in chronic obstructive lung diseases]. AB - This study was conducted to investigate the clinical value of blood gas analysis during exercise in patients with COPD and healthy controls using the new criteria of the Deutsche Gesellschaft fur Pneumologie (DGP) for performance and interpretation of blood gas analysis during exercise. A total of 64 patients with COPD (age: 63.7 +/- 10.1 years) and 35 healthy controls (age: 35.0 +/- 14.3 years) exercised on a bicycle ergometer to their submaximal capacity under steady state conditions (patients: 44.4 +/- 24.6 watt vs. 102.4 +/- 32.1 watt; p < 0.01). A pathological response for PaO2 occurred in 3 of 35 controls (8.6%) and 14 of 64 patients (21.9%; p < 0.01). A decrease in PaO2 yielded a sensitivity of 28% and a specificity of 92% in respect of patients with emphysema. Significant associations of lung function parameters at rest and blood gases under exercise were found by linear regression analysis (airway resistance (Rt): beta = -0.48; p < 0.001) and PaCO2 (FEV1: beta = -0.27; p < 0.05). This study demonstrates that in patients with COPD parameters of lung function and blood gas analysis at rest are already good predictors of gas exchange under exercise conditions. In individual cases, however, prediction may not be possible. This underlines the importance of the exercise test to investigate gas exchange under diagnostic (i.e. dyspnoea, medical opinion) and therapeutic aspects (i.e. therapy control). For COPD patients, the rule of the DGP yielded a poor sensitivity, but an excellent specificity in the diagnosis of emphysema, which yields confirmation of the diagnosis only in conjunction with, and complementary to, other methods. PMID- 9333801 TI - [Pulmonary tumor cell embolisms as primary manifestation of malignant neoplasms. Review of incidence, morphologic and clinical findings of a disease picture rarely diagnosed before death]. PMID- 9333802 TI - [Anti-leukotrienes--a new perspective in treatment of bronchial asthma. 1. Scientific Pneumology Symposium, Cannes, 14-16 February 1997]. PMID- 9333803 TI - [Complicated course of rheumatoid arthritis with pulmonary involvement, myocardial fibrosis and sleep apnea syndrome]. PMID- 9333804 TI - [Complicated course of rheumatoid arthritis with pulmonary involvement, myocardial fibrosis and sleep apnea syndrome]. PMID- 9333805 TI - [Analysis of tuberculosis mortality dynamics in Sverdlovsk region in 1995-96]. AB - Among the deceased in the Sverdlovsk Region in 1995 - 1996 there was a larger proportion of males and individuals aged over 40 years, unemployed, homeless persons, patients with infiltrative pulmonary tuberculosis, concomitant diseases and a smaller proportion of those with prolonged pulmonary tuberculosis, its fibrocavernous type, persons who died from secondary nonspecific changes and complications of the tuberculosis process. Today the most common causes of premature death due to tuberculosis are in patients' refusal of treatment, systemic incompliance, a severe concomitant disease, contraindications for surgical treatment, drug resistance to tuberculostatic agents, drug shortage, late referral for medical aid and long-term evasion of prophylactic surveys for tuberculosis. PMID- 9333806 TI - [Emergency diagnosis of tuberculosis in patients admitted to the Institute of Emergency Care]. AB - Tuberculosis was first recognized by a phthisiothera-pist in 52 patients and in 3 cases at autopsy at the admission and resuscitation units. Out of 55 patients, 39 (71 +/- 6.1%) were found to have acute, general processes (subacute dissemination and acute military tuberculosis, caseous pneumonia, multiple preliminary extrapulmonary diseases). Six patients died at the Institute, after referral to tuberculosis dispensaries, there were 11 and 9 deaths at months 1 and 6, respectively. Mortality was 47.3 +/- 6.7%. According to the data of emergency x ray study, the diagnosis to be correct in 87.3 +/- 4.2%. As high as 25% of new cases of tuberculosis were detected at somatic hospitals in 1992 - 1994. Therefore, all patients admitted for pulmonary complaints of general toxic syndrome should immediately undergo X-ray, sputum test for Mycobacterium tuberculosis, and a phthisiotherapist's examination. PMID- 9333807 TI - [Problems in the treatment of patients with pulmonary tuberculosis]. AB - The paper presents the current approaches to chemotherapy in patients with pulmonary tuberculosis and shows the main reasons that do not allow one to achieve high outcomes of treatment. These involve the drug resistance of Mycobacterium tuberculosis, the morphological features of a specific process in the lung, the higher incidence of acutely progressive types of pulmonary tuberculosis in particular; tuberculosis-contaminant diseases (diabetes mellitus, gastrointestinal, hepatic, renal diseases, and non-specific respiratory diseases, etc.). Recommendations how to eliminate the reasons for ineffective treatment of pulmonary tuberculosis are given. PMID- 9333808 TI - [Use of maxaquin in the treatment of progressive pulmonary tuberculosis occurring with standard chemotherapy regimens]. AB - To study the efficacy of the fluoroquinolone maxaquin (lomefloxacin), the latter was used in 27 patients with destructive bacillary pulmonary tuberculosis which progressed with routine drug regimens. After monthly therapy, positive clinical changes were achieved in 85.6% of patients, abacillation was obtained in 33.2%, a X-ray decrease in infiltrative and focal changes was recorded in 81.5% and decay cavitary closure occurred in 22.2% of patients at month 3 of maxaquin therapy. The positive clinical, laboratory, and X-ray changes occurring after the use of maxaquin necessitates the supplementation of this agent to the drug regimen for patients with progressive pulmonary tuberculosis. Mycobacterial resistance to the major drugs is an indication for the use of maxaquin. PMID- 9333809 TI - [Treatment effectiveness of patients with ocular tuberculosis at specialized sanatoria]. AB - The outcomes of treatment of 2908 patients with ocular tuberculosis who were treated at the Vyborg-3, Krasnyi Val, and Plyos sanatoria in 1993 - 1995 are analyzed. These sanatoria used the common guide in evaluating the efficacy of treatment, namely a separate assessment of the outcomes of treatment of the underlying and concurrent diseases. There were no statistical significant differences in therapeutical efficiency at these sanatoria. Therapeutical benefits were obtained in 99.8% of patients in the active phase of the disease. PMID- 9333810 TI - [Use of immunochemical studies to predict the course of fibrous cavernous tuberculosis of lung and postoperative complications in patients on chemo and laser therapy]. AB - A total of 103 patients with fibrocavernous tuberculosis of the lung were examined. They all received chemotherapy, including 3 - 4 antituberculous agents. Laser therapy was performed with a UZOR-2K low-energy semiconductor laser. In patients with profound changes in the serum level of protein, with high antigenemia and antibody production, the course of the disease was found to be poor; X-ray positive changes were achieved to a lesser extent, bacterial expellation stopped less frequently and more slowly. The decreases in the serum content of the proteins tested, in the level of antigenemia and antibody production which occur with drug and laser therapies are also an important factor of preoperative preparation, which is highly effective in preventing postoperative complications. PMID- 9333811 TI - [Mycobacterial antigens and antitubercular antibodies in patients with tuberculosis salpingo-oophoritis]. AB - Sixty two and 59 patients with tuberculous and nontuberculous salpingo oophoritis, respectively, and 30 healthy females were examined. It was found that indirect solid-phase enzyme immunoassay (EIA) using ultrasonography of Mycobacterium tuberculosis H37Rv could detect antituberculosis antibodies in 66.7% of patients with tuberculous salpingo-oophoritis, and in 10% of healthy females. That using the antigen isolated from the cell wall extract of M. tuberculosis (whose molecular weight was 38 - 42 kD) could reveal them in 70.2, 4.3%, and 6.7%, respectively. After dissociation of immune complexes, EIA inhibition using affinity-purified antimycobacterial antisera displayed mycobacterial antigens in 75.0 and 4.3% of patients with tuberculous and nontuberculous salpingo-oophoritis, respectively, and in none healthy female. Thus, the detection of mycobacterial antigens and antituberculosis antibody may be successfully used in the complex diagnosis of tuberculosis of the female genitals. PMID- 9333812 TI - [Detection of antitubercular antibodies in children and adolescents at the health resort stage of management]. AB - Antituberculosis antibodies were detected in 164 children and adolescents who had tuberculosis and who were infected with mycobacteria. The diagnostic value of indirect hamagglutination tests with phosphatide antigen and tuberculin, complement uptake, passive hemolysis, and enzyme immunoassay for evaluating the progression of tuberculosis was defined in patients at a specialized sanatorium. The significance of the tests increases when they are concurrently used. PMID- 9333813 TI - [Tuberculosis of large bronchi, specific features of course and its clinical significance]. AB - The author analyzed the outcomes of treatment of 88 patients with various types of pulmonary tuberculosis, of them 44 (Group 1) were diagnosed as having tuberculosis of the large bronchi (TLB) by bronchoscopy and verified by cytological of morphological studies of biopsy specimens and the other 44 (Group 2) had intact bronchi. TLB occurred in all age groups and in various types of the pulmonary tuberculosis. This complication occurred at the patients" late referral and in the presence of other complications and concomitant diseases. The patients with TLB were found to have slow rates of abacillation and decay cavity closure in the lung, which required more prolonged treatment. PMID- 9333814 TI - [Prevalence and specific features of pulmonary emphysema in patients with pulmonary tuberculosis]. AB - The prevalence and specific features of emphysema of the lung were studied in patients with pulmonary tuberculosis. The temporal pattern of the process and secondary obstructive genesis were established. It was stated that there was a redistribution of the existing emphysematous changes into the unaffected parts of the lung along with the higher scope of the chronic tuberculosis process. PMID- 9333815 TI - [Effect of phytotherapy on the prevention and elimination of hepatotoxic responses in patients with pulmonary tuberculosis, carriers of hepatitis B virus markers]. AB - Whether hepatotoxic responses (HTR) can be prevented and corrected in patients with pulmonary tuberculosis, carriers of hepatitis B virus markers during specific therapy by using preventive hepatoprotective phytotherapy was studied. Group 1 (n = 54) was given individually chosen plant species infusions having a pronounced heptoprotective effect (liquorice (Glycyrrhiza L.), nettle (Urita L.), tansy (Tanacotum L.), mint (Mentha L.), etc.)) in the whole course of antituberculous treatment. A control group (n = 58) received essentiale, legilon, Liv-52, hemodez from the onset of HTR to their elimination. Phytotherapy was found to reduce to occurrence of HTR by 4.1 times, to eliminate them 2.1 times more rapidly, to shorten the periods of discontinuation of antituberculous agents by 2.1 times, which positively affects the time and efficiency of treatment for tuberculosis. PMID- 9333816 TI - [Laser therapy in patients with inflammatory pleural exudates]. AB - The outcomes of treatment of 140 patients with tuberculous parapneumatic serous pleurisy are presented, Intrapleural low-energy laser radiation, epicutaneous low energy pulse radiation, and the successive application of both techniques were used during etiotropic therapy and regular exudate aspirations. There was reduction in the time of exudation into the pleural cavity, as well as recovery with less pronounced pleural impositions. PMID- 9333818 TI - [Biorhythm of hormonal system in chronic bronchitis]. AB - In the first 3 days, circadian biological hormonal variations were studied 5 times a day (at 8, 12, 16, 20, and 24 o'clock in 103 patients with chronic bronchitis, including 41 and 62 with nonobstructive and obstructive bronchitis, respectively. Hormonal changes in chronic bronchitis were evaluated as asynchronism manifested by the time displacement of acrophases, the imbalance of mesor and the amplitude of diurnal hormone level variations, the intrinsic discrepancy of linkages in the horizontal lines and endocrine functional changes. PMID- 9333817 TI - [Organization of in- and outpatient treatment of new cases of destructive pulmonary tuberculosis]. AB - A system of in- and outpatient treatment of the first detected patients with destructive pulmonary tuberculosis is proposed. The use of various types of intravenous intermittent chemotherapy in patients with focal, infiltrative, and disseminated tuberculosis provides a high efficiency in the inpatient period (the mean time of sputum negativation is 1.6 months, that of destruction closure is 3.1 months) and early (in 2 - 3 months) transfer of these patients to outpatient additional treatment by continuing etiotropic therapy that has turned out to be effective in hospital. PMID- 9333819 TI - [Tuberculous lymphadenitis as a topical problem of phthisiology]. AB - The paper presents the results of follow-up of 894 patients in 1983 - 1993. The increasing topicality for physiology and the potentialities of making the diagnosis of the disease better are defined, based on the data on the pathogenesis and pathomorphism of tuberculous lymphadenitis and on the results of testing new diagnostic techniques, including cytological and histological studies of sections and imprints, enzyme immunoassay, and polymerase chain reaction. PMID- 9333820 TI - [Tissue destruction and initiation of blood coagulation in tuberculous inflammation]. PMID- 9333821 TI - [Immunological aspects of tuberculosis uveitis pathogenesis]. AB - Based on clinical and immunological examinations of 251 patients with tuberculous uveitis (TU), 3 types of its course (productive, exudative, and mixed) were identified. The immunopathologic pattern of each type was found to be different: the productive type was characterized by delayed hypersensitivity, the exudative one had immediate hypersensitivity, and the mixed one had both. The immunogenetic markers of mixed TU were defined. These included HLA antigens, such as A2, B7, B21, and B27. PMID- 9333822 TI - [Feasibility of using gas-liquid chromatography to identify microbes of Nocardia genus]. AB - The use of gas-liquid chromatography based on the determination of the fatty acid composition of bacterial cell makes it possible to rapidly identify Nocardia isolated from the patients' smears. Examining the fatty acid composition of museum and fresh Nocardia cultures suggests that they are largely homogeneous, The major fatty acids in Nocarida are hexadecenoic, octadecenoic, and tuberculostearic acids. Criteria for differentiating bacteria of the genus Nocarida via gas chromatographic analysis of their fatty acid spectrum from Mycobacteria and other nonmycobacterial acid-resistant microbes. PMID- 9333823 TI - [Pulmonary tuberculosis and acquired immunodeficiency syndrome in Ukraine (first communication)]. AB - The epidemiological HIV infection situation in the Ukraine in the past 4 years is analyzed. In patients with AIDS, the clinical manifestations of tuberculosis are atypical; prolonged intoxication, negative tuberculin tests, no bacterial isolation in half the cases. These patients should be treated with 4 - 5 antituberculous drugs, by taking into account the fact that only long-term continuous therapy may yield a positive effect. PMID- 9333824 TI - [Impact of low-energy laser irradiation on the immunological nonspecific responsiveness in patients with tuberculosis]. PMID- 9333825 TI - [Recurrence of pulmonary tuberculosis: in- and outpatient data]. AB - Diagnosis recrudescent pulmonary tuberculosis shows different flaws: on the one hand, the progression of early untreated tuberculosis can be mistaken for a recrudescence, on the other, changes are interpreted as a recrudescence if there is no clinical and X-ray evidence for the reactivation of the process. All cases of recrudescent tuberculosis should be reexamined by a medical commission, which allows the above errors to be avoided. If a patient with chronic alcoholism and mental disease has recrudescent tuberculosis, a narcologist and psychiatrist must participate in working out a comprehensive treatment policy. PMID- 9333826 TI - [Current methods for treating pulmonary tuberculosis (review of 1994-1995 literature)]. PMID- 9333827 TI - [Integral factorial assessment of preventive measure system in tuberculosis]. AB - The antituberculosis measures made have achieved their peak efficiency and, if specific measures to increase the detection rates of patients in the general population and the level of vaccination cannot be found now, tuberculosis morbidity rates cannot be expected to become lower in the near future. This is evidenced by the 1996 prognosis that the morbidity will rise from 84.6 to 96.1 per 10,000 persons. It is expedient to implement preventive antituberculosis measures, by taking into account the regional features of a specific area. PMID- 9333828 TI - [Impact of family contact on development of tuberculosis in infants and preschool children]. AB - The paper outlines the results of following up 50 infants and preschool children who had contacts on their families. The major sources of infection were equally both their mother and their father. The children were found to have pronounced active processes. The most severe types of tuberculosis were common in infants in whom general, military tuberculosis and tuberculous meningitis along with complicated types in intrathoracic lymph node tuberculosis were detected. There was more commonly a phase of dissemination and decay in infants. Severe types of tuberculosis were encountered in the families having a combination of many unfavourable factors, both medical and social ones. Among the children who had fallen ill, 32% were detected on their visits to polyclinics of upon admissions to general hospitals. PMID- 9333829 TI - [Personal values and goal orientations of chronically ill adolescents and young adults]. AB - For all those involved in the care of the chronically ill, to provide support in a professional manner implies to strive for a comprehensive understanding of the specific coping strategies in the light of the individual person's priorities. Coping research has yet to present a systematic analysis of personal values, goals and expectations of the chronically ill. The reader is invited to approach the topic by a review of the relevant literature and presentation of our own study on the classification of individual values and goals in 164 chronically ill adolescents and young adults (cystic fibrosis, Crohn's disease, haemophilia). Regarding personal values, the responses by the young patients were allocated to three main categories: pursuit of social contacts, striving for acceptance of the disease, the striving for joyful and intense living. Finally the relevance of personal values for appropriately coping with a serious illness is discussed. PMID- 9333830 TI - [Confounding effects in social science studies and their control. The examples of personality, social support, psychiatric disease]. AB - Social scientific investigations on coping with illness, on the relevance of life events or social support for psychic disorders are concerned with a variety of constructs with interdependent structures. Hypotheses in interrelationships are set up and tested on the construct level. A problem quite often overlooked here is that almost all constructs overlap to a considerable extent both with regard to their content as well as operationally, i.e. they are confounded. Interrelations of constructs which are strongly content-related or operationally confounded are thus in danger of demonstrating artificial results. Data from an extensive epidemiological research project on the incidence and prevalence of psychic disorders in the general population are used applying structural equation models for latent variables to demonstrate confoundation effects for the constructs personality, social support and state of health. Appropriate control methods are described. The results show that partner support maintains a strong influence on the psychic and physical state of health even confoundation of content has been excluded. Application of this method of content confoundation control is discussed with regard to research on coping life event research etc. PMID- 9333831 TI - [Do psychological factors modify survival of cancer patients? II: Results of an empirical study with bronchial carcinoma patients]. AB - The present prospective test study of hypotheses addressed the question whether psychological factors are predictive of survival time in lung cancer patients. The hypotheses were: Emotional distress, depression and depressive coping are associated with shorter survival; hope and active coping with longer survival. The study was based on a sample of n = 103 patients who were investigated post diagnosis and before the beginning of primary treatment. Emotional distress and hope were assessed by clinical scales (self-reports and interviewer ratings), depression by the Depression Scale of von Zerssen, depressive coping and active coping by the Freiburg Questionnaire on Coping with illness by Muthny. At follow up, which took place three to five years later, n = 74 patients had died, for n = 29 patients the survival data are censored. The prediction of the survival time was performed applying multivariate analyses (Kaplan-Meier-method, Cox Regression), adjusting for biological risk factors (histological classification, stage of the disease, type and amount of treatment, Karnofsky performance status, age). Results were as follows: Active coping and hope were associated with longer survival, emotional distress, depression and depressive coping with shorter survival, respectively. These associations were found consistently across assessment methods. The predictive effects of coping and distress were statistically independent of the influence of the somatic risk factors. The best psychological predictor was the interviewer rating of active coping. Its predictive power equalled that of the Karnofsky performance status. However, there was evidence that the effects of the psychological factors varied somewhat in interaction with treatment modalities. The findings are discussed from a methodological perspective. Possible causal models and mechanisms are presented which could account for interactions of psychological measures and the course of the disease: Thus, it can be conceived that psychological effects were mediated by patients' compliance with medical treatment. In addition, it cannot be ruled out that psychological factors themselves were influenced by the physical status of the patients at the time of entry to the study. PMID- 9333833 TI - [Anxiety as an existential phenomenon. An existential analytic approach to understanding and therapy of anxieties]. AB - From the existential analytical point of view, anxiety is considered to be a basic theme of existence. The experience of being threatened is most commonly related to the physical and material aspects of life. But on a deeper level anxiety deals with the search for foundational and supporting structures for existence. When one loses the sense of safety of being held and of having shelter in a world that does not offer ultimate securities, one is prone to anxiety. Anxiety can therefore be perceived as a subjective parameter of feeling threatened in the existential structures. This paper gives an outline of an existential analytical approach to the understanding of fear and anxiety, a basic classification of anxieties, and an overview of the following specific methods of an existential treatment search for foundational structures of existence, personal position-taking, dereflexion (Frankl) and paradoxical intention (Frankl). PMID- 9333834 TI - [Utilization of ambulatory psychotherapy by patients with endogenous depression after inpatient psychiatric treatment]. AB - The long-term course of major depressive disorder is often accompanied by relapses or chronicity. Since psychosocial factors have been shown to be important predictors for the long-term outcome, psychotherapy along with drug therapy belongs to the standard methods of treatment. In spite of the effectiveness of psychotherapy, only some of the inpatients are treated with outpatient psychotherapy after hospital discharge. Within the framework of the Heidelberg depression study the authors examined what kind of, how many and for how long endogenously depressed patients sought out-patient psychotherapy after an inpatient treatment in a two-year follow-up. During the follow-up one half of patients were treated with outpatient psychotherapy. They were found to be younger, had suffered from more previous episodes, and their personality was more disturbed than those who had not undergone psychotherapy. The Expressed Emotion index did not make any further difference, whereas certain aspects of partnership quality did. The distinction between the two groups of patients is discussed regarding possible selection processes by treatment indication. It is pointed out that research on synergistic cooperation between members of the mental health services is highly desirable. PMID- 9333832 TI - [From hysteria to nervousness. Comments on Willy Hellpach's sociopathologic prognoses for the 20th century]. AB - The very rich, extensive, and complex work of the psychologist, doctor of medicine, and politician Willy Hellpach (1877-1955) is largely forgotten today. Our article sheds light upon his contribution to the nosology of neuroses, which emphasizes the impact of history, society, and culture on the neurotic syndromes. Hellpach states that during the process of civilisation hysteria is more and more replaced by nervousness. In contemporary nosology this hypothesis leads to the discussion how the relationship of neuroses with depressive symptomatology on the one hand and neuroses with panic attacks and phobia on the other should be conceptualised. PMID- 9333836 TI - [Quality of life with intensive insulin therapy: a prospective comparison of insulin pen and pump]. AB - The purpose of the following-study was to identify aspects of quality of life that are particularly affected by the mode of insulin therapy. 55 patients with insulin-dependent diabetes mellitus, who volunteered for a change of their intensive insulin therapy with pen injections to continuous subcutaneous insulin infusion (CSII) were studied 1 month before, and 6 months after, changing to CSII. The DCCT questionnaire was applied, measuring quality of life in the 4 subscales: satisfaction, impact, social/vocational worries, and diabetes related worries, respectively. The results demonstrate that the "satisfaction" subscale was scored significantly higher (p < 0.02), and the "impact" subscale was scored lower (p < 0.02) with CSII therapy. Single items showed that this was due to greater flexibility with leisure-time activities and with diet, and to significantly less problems with hypoglycaemia. The subscales "social/vocational worries" and "diabetes-related worries" were scored unchanged, HbA1c changed only slightly from 7.5% (SD 1.2) to 6.9% (SD 0.9): (p < 0.05). It is concluded that disease-related deficiencies in quality of life (satisfaction, impact) improve considerably in insulin-dependent diabetic patients after changing voluntarily from intensive insulin therapy with pen injections to continuous subcutaneous insulin infusion. PMID- 9333835 TI - [Age dependence of coping strategies in children and adolescents with diabetes mellitus]. AB - In this cross-sectional study the age-dependency of coping behaviours of 43 children and adolescents with IDDM aged 8 to 18 years has been assessed by means of the German version of the Kidcope (Rathner u. Zangerle 1996). Everyday stressors as well as disease-related stressors were used. Significant age dependent differences in the frequency of coping behaviours could be found in all strategies except self-criticism. Especially distraction and wishful thinking were significantly more often used by children than by adolescents, whereas adolescent used significantly more often resignation. The more frequent use of resignation by adolescents is not due to a longer duration of illness, but to their developmental stage. Thus, in chronically ill children and adolescents developmental aspects showed a stronger influence on coping than duration of illness. Self-criticism, blaming of others and social withdrawal are as rarely used by children as by adolescents. Self-rated efficacy of coping strategies showed almost no age differences; social support was rated as the most efficient coping strategy by both age groups. Resignation, in the sense of a cognitive acceptance of the illness, seems to be more efficient for adolescents than for children. It is concluded that the developmental process of children and adolescents with chronic illnesses such as IDDM may lead to a better cognitive acceptance of the disease. PMID- 9333837 TI - [HyperPsych--resources for medicine and psychology on the World Wide Web]. AB - Progress in the research of interactive communication technology and the acceleration of processing and transmitting information have promoted the development of computer networks allowing global access to scientific information and services. The recently most well-known net is the internet. Based on its integrative structure as a communication-directed as well as an information directed medium, the internet helps researchers design scientific research. Especially medicine and psychology as information-dependent scientific disciplines may profit by using this technological offer. As a method to coordinate to the vast amount of medical and psychological data around the globe and to communicate with researchers world-wide, it enhances innovative possibilities for research, diagnosis and therapy. Currently, the World Wide Web is regarded as the most user-friendly and practical of all the internet resources. Based on a systematic introduction to the applications of the WWW, this article discusses relevant resources, points out possibilities and limits of network-supported scientific research and proposes many uses of this new medium. PMID- 9333838 TI - [The prevalence of Ixodes ricinus ticks (Acari, Ixodidae) in the forested areas of Gdansk, Sopot, and Gdynia and their infection rate with Borrelia burgdorferi spirochetes]. AB - Data are presented on the variable patterns on the seasonal activity of Ixodes ricinus questing on vegetation in 6 study sites in the forested areas of Gdansk, Sopot and Gdynia in 1993-1995. A total of 8992 specimens collected there show that ticks frequently occupy habitats closely associated with man. Out of them 5775 (4328 nymphs, 713 females and 680 males) collected in 1994 and 1995 were examined individually for Borrelia burgdorferi sensu lato-the etiologic agent of Lyme borreliosis-using in-direct immunofluorescence assay (IFA). Spirochetes were detected in 577 (10.3%) of the ticks tested. The overall infection rate was 8.2% for nymphs (n = 353), 14.9% for females (n = 102) and 18.9% for males (n = 122). The infection rates in particular study sites varied between 1.4% and 16.4% in 1994 and between 3.6% and 26.9% in 1995. The highest prevalence of B. burgdorferi was observed in June (10%) and October (11.9%) in 1994 and in August (16.3%) and October (25%) in 1995. Detection of B. burgdorferi in ticks derived from the same area in the two following years shows that the infection of the I. ricinus population with this pathogen in the forested areas of Gdansk, Sopot and Gdynia is permanent. PMID- 9333839 TI - [Botulism in 1995]. PMID- 9333840 TI - [Human poisonings caused by agents used for plant protection in 1995]. PMID- 9333841 TI - [Viral hepatitis B in 1995]. PMID- 9333842 TI - [Viral hepatitis non-B in 1995]. PMID- 9333844 TI - [Rabies in 1995]. PMID- 9333843 TI - [Tetanus in 1995]. PMID- 9333845 TI - [Brucellosis in 1995]. PMID- 9333846 TI - [Trichinosis in 1995]. PMID- 9333847 TI - [Taeniasis in 1995]. PMID- 9333848 TI - [AIDS and HIV infection in 1995]. PMID- 9333849 TI - [Malaria in Poland]. AB - Malaria epidemiological situation in Poland since nineteenth century to 1995 has been described. The changes observed during this period are enormous. Poland has been transformed from endemic country with huge epidemics into the country with sporadic imported malaria cases. PMID- 9333850 TI - [Territorial distribution of neural tube defects in Poland]. AB - Neural tube defects (ntd) remain unresolved therapeutic problems, even for contemporary health services. Prophylactic measures are the best methods for limitation of the problem. In many countries, the number of children born with ntd has been reduced significantly, even in 4-5 folds. Authors found that the situation in Poland remains stable since last 20 years. Epidemiological evaluation of the ntd morbidity in Poland is very difficult. There is no National Register of Inherited Defects. Authors observed and analysed mortality and hospitalisation due to ntd in Poland, in early ninetees, and compared these data with information originated from Regional (Wielkopolska) Inherited Defects Register. It was found that ntd mortality and hospitalisation in Wielkopolska was similar to average in Poland. It has been estimated that the rate of children born with ntd in Poland, in 1995 was 2.68 per 1000 live and stillbriths (in 1972 1974 was 2.04). Especially high rates of ntd's have been found in north-eastern part of Country. Incidence of ntd's was higher in rural, than in urban population of the Country. PMID- 9333852 TI - [Program contents of undergraduate teaching of epidemiology to medical students and its evaluation]. AB - The importance of epidemiology in modern medicine increased rapidly during the last years. Now each physician should know basic epidemiological definitions and epidemiological strategy for studies on health and disease. The course of epidemiology, as a separate subject for medical students, was introduced to curriculum of Medical Faculty in 1970. At present, the course covers 4 lecture units and 16 exercise units. It is conducted on 4th year of the Medical Faculty. The evaluation of the course carried out in 1995 showed that a great majority of medical students considers it as interesting and useful subject to be taught. PMID- 9333851 TI - [Classification exposure error in studies on tobacco smoke in pregnancy and birth weight of newborns]. AB - The purpose of the paper was to assess the validity of the self-reported tobacco smoke exposure in pregnancy against plasma cotinine measurements. A total of 158 patients from obstetrical wards was included in the cotinine study. Biochemical smokers were defined as persons with serum cotinine levels greater than 25 ng/ml. The data showed that the exposure classification based on self-reported smoking habit status confronted with cotinine values was of low sensitivity (52%) but of high specificity (98%). To assess the effect of this exposure classification error on the association between the low birth weight (LBW) and smoking in pregnancy, the data from the recent survey in Cracow children have been used. It was shown that RR estimates for smoking and LBW after adjustment for misclassification error were substantially higher than that not adjusted (crude RR = 2.9, corrected RR = 5.1). Due to the exposure misclassification error the attributable fraction (AF) of LBW due to mother's smoking was heavily biased as well. Estimated attributable fraction AF(pop) based on crude RR amounted to 22%, however, after adjustment reached 50%. The corresponding values for attributable fraction in exposed group AF(exp) were 66% and 80%. PMID- 9333853 TI - [The evaluation of the professional training of physicians specializing in the epidemiology of infectious diseases. A set of examination questions]. PMID- 9333854 TI - [Plasma levels of interferon gamma and interleukin 10 in patients with lymphonodular toxoplasmosis]. AB - The concentrations of IFN gamma and IL 10 in plasma of sixteen patients with toxoplasmic lymphadenopathy were measured. These examinations were carried out two times in the interval of a month. We found increased level of IFN gamma and normal concentrations of IL 10 in both of these terms. PMID- 9333855 TI - [Report from "Zalzburg-Cornell Seminars", Salzburg, July 1996]. PMID- 9333856 TI - [Diphtheria and tetanus immunity of blood donors]. AB - Immunity to diphtheria and tetanus was determined in serum samples from 108 blood donors. Antitoxin concentration was evaluated by ELISA assay. 65% of donors were protected to diptheria and 100% to tetanus. Significant trend of decreasing immunity was observed with increasing age. PMID- 9333857 TI - [Infectious diseases in Poland in 1995]. PMID- 9333858 TI - [The epidemiology and laboratory diagnosis of Pneumocystis carinii infections]. PMID- 9333859 TI - [Measles in 1995]. PMID- 9333860 TI - [Whooping cough in 1995]. PMID- 9333861 TI - [Scarlet fever in 1995]. PMID- 9333862 TI - [Mumps in 1995]. PMID- 9333863 TI - [Influenza in 1995]. PMID- 9333864 TI - [Rubella in 1995]. PMID- 9333865 TI - [Cerebrospinal meningitis and encephalitis in 1995]. PMID- 9333866 TI - [Salmonellosis in 1995]. PMID- 9333867 TI - [Bacterial dysentery in 1995]. PMID- 9333868 TI - [Food poisoning and infection in 1995]. PMID- 9333869 TI - [Clinical diagnosis of osteoporosis]. AB - Osteoporosis (OP) is a systemic heterogenic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue with a consequent increase in bone fragility and susceptibility to fractures. The fractures are the clinical consequences of the disease. Before fractures occur there is a long period without clinical significance characterised only by osteopenia. Both sexes lose bone mass during lifetime but after menopause women lose bone much faster and are more prone to fragility fractures. Wrist fractures (Colles fractures) occur in the earliest postmenopausal period in the fifth decade of a woman's life, and these fractures are 7 times more frequent in women than in men. Vertebral fractures occur 10-15 years after the menopause. Only 10-20% of these cases are hospitalized because of pain, and up to 80% of the cases are clinically symptomless. Kyphosis and loss of height come in later years. The most serious fractures are those of the hip. They usually occur over the age of 70 and bring 8 10% hospital mortality, about 20% mortality in the following year, and 50% disability over the age of 80. The diagnosis of primary OP requires the exclusion of all possible secondary causes of OP. In the diagnosis of OP a medical history with risk factor of OP is very important, as well as physical examination, and some laboratory tests. X-ray bone evaluation and the bone densitometric measurements are very important in confirming of osteopenia and osteoporosis. PMID- 9333870 TI - [Radiologic diagnosis and densitometry of osteoporosis]. AB - The principles of densitometric and radiological examination in diagnosing of osteoporosis are presented. Peripheral and whole body densitometers are described and their quality is assessed on basis of precision and accuracy of bone mineral density measurements (BMD). WHO criteria of diagnosis, which introduce clinical category of the basis of BMD (expressed in T-scores) are described as follows: normal-(T-score up to -1.0); osteopenia-(T-score below -1.0 but more than -2.5); osteoporosis-(T-score -2.5 or below); severe osteoporosis-(T-score below -2.5 and one or more fractures). Evaluation of osteoporotic changes on the spine, pelvis and hand radiographs are discussed. Quantitative and possible quantitative analysis of radiographs are described. PMID- 9333871 TI - New perspectives on bone densitometry. AB - Methods of bone mineral density measurement are presented. Radiological photogrammetry, single photon absorptiometry, dual photon absorptiometry and dual energy X-ray absorptiometry are described. Mode of work of densitometers and their characteristics ares presented. Modern development of densitometric equipment and possible application in diagnosing of osteoporosis is discussed. PMID- 9333872 TI - [Physical aspects of selected densitometry methods used in the diagnosis of osteoporosis]. AB - Physical background of diagnostic procedures currently applied to assess bone mineral density (single and dual X-ray absorptiometry, quantitative computed tomography and ultrasonography) are described. Special emphasis is given to physical interpretation of diagnostic parameters and to the comparison of the procedures. Systematic errors of the procedures is discussed in detail. PMID- 9333873 TI - [Examination of the mechanical properties of bone]. AB - The skeletal system from mechanical point of view operates as supporting structure carrying forces and moments which occur during regular functioning of human body. The disturbances of these functions results in changing of mechanical characteristics of bones tissue so their evaluations could be a useful indicator of sickness and pathological conditions. The research of methods of mechanical characteristics of biological tissue should pay an attention to its strong anisotropy and sensitivity with respect to the conditions of the examination. A model of paradigm in biomechanics bones was described with its specifics and conditions. The theoretical description of the mechanics of bones deals with constitutive equations corresponding to the functional Wolff's adaptation theory. For proper estimation of strength the laws of crack propagation were applied. Some methods on mechanical and age-related characteristic, were described. As an example research on bone protection in the shoulder area against the results of side hitting (e.g. car accidents) was presented. This paper shows that bone mechanics study enables a better understanding of the relations between the mechanical properties of bones and the tendencies to illness and their changes with biological age. PMID- 9333874 TI - [Usefulness of bone densitometry, quantitative computed tomography and quantitative ultrasound in diagnosis and monitoring of osteoporosis treatment]. AB - Bone densitometry is characterized by high sensitivity and specificity in osteoporosis, and new generations of densitometers enable measurements with improved intra- and inter-assay precision. The clinical potential of bone densitometry is well documented and the technique is widely used in clinical practice. It does not, however, allow for measurement of "true" bone density; instead it measures so called serial density (expressed in g/cm2) which is the distribution of bone mass over the flat projection of the skeleton. Limitations of densitometric techniques can be overcome by applying other methods, i.e. quantitative computed tomography (QCT) and ultrasound (US). QCT enables separate measurements of compact and trabecular bone density (expressed in g/cm3), as well as calculation of Strength-Strain Index (SSI), reflecting the mechanical resistance of bone to fracture. US is a non-invasive technique, providing information of fracture risk and bone tissue quality. Both techniques seem very promising and have been extensively studied recently; they are expected to move from clinical research to clinical practice soon. PMID- 9333875 TI - [Difficulties in the interpretation of bone densitometric measurements]. AB - In evaluation of the densitometric measurements the most popular presentation of results is a form of t-score or z-score values. T-score is a result of bone mineral density (BMD) in SD below a peak bone mass and z-score is a figure in SD below the values of BMD of healthy individuals of the same age and sex. Osteoporosis is diagnosed if t-score is more than -2.5 ranges from -1.0 to -2.5. Z-score has less value in diagnosis of osteoporosis. The value of different densitometric techniques depends on age of the subject and a past history of fractures. In subjects below 65 yrs the best technique is DEXA (dual energy X ray absorptiometry) and the spine is the site of measurement. In subjects above 65 yrs the best site to measure BMD is the hip. Previous fractures at the site of measurement (vertebral crush fractures), osteoarthritis, kyphosis, could change the values of BMD (overestimate them), and there is a need for a very careful and critical interpretation of BMD results. PMID- 9333876 TI - [Computer analysis of the radiographic image of bones and joints]. AB - Various possibility of computerized analysis of radiographic image of bone and joint are presented. The principle of digital image recording and processing are described. Practical application of computer programs of bone structure analysis and measurement of joint width using image analysis procedure are presented. PMID- 9333877 TI - [Differential diagnosis of osteoporosis]. AB - Differentiation of different types of osteoporosis e.g. postmenopausal and senile is presented as well as differentiation of osteoporosis and osteomalacia. Differential diagnosis between osteoporosis and Paget's disease is performed. PMID- 9333878 TI - [Evaluation of the prevalence of osteoporosis in a population of women living in Krakow based on densitometric measurements of the forearm]. AB - Osteoporosis is considered a civilization disease, affecting populations living in big cities. It is detected in 10 percent of population and in 30 percent women over fifty. We tried to estimate prevalence of osteoporosis in people living in Krakow. For estimation purposes, 1000 people living in Krakow more than 40 years were chosen. 325 persons came to our Department for examination, 232 of which were women. All patient filled a special questionnaire on pathological changes in skeletal system and risk factor of osteoporosis. Densitometric measurements of nondominant forearm using Osteometer DTX 100 were performed in all patients. For osteoporosis criteria level of T-score less than -2.5 was taken. Because of lack of Polish reference data for densitometric measurements our data were compared to WHO data, concerning prevalence of osteoporosis in white women. Compared to WHO data, a higher frequency of osteoporosis was found in women living in Krakow more than 40 years. PMID- 9333879 TI - [Densitometric evaluation of the distal radial epiphysis in patients after Colles fracture]. AB - The results of bone mineral density measurements in both forearms of 50 patients aged from 51 to 83 yrs (av. 62.4) who sustained unilateral Colles fracture, were analysed. Time elapsed from fracture was from 0.5 to 6 yrs. The aim of this study was to assess the effect of previous Colles fracture on bone mineral density. It was found that after the preliminary increase of bone mineral density, there was a gradual decrease towards normal values as compared with the unbroken arm. The bone mineral density in the evaluated group with fracture was significantly lower than the control groups without fracture. PMID- 9333880 TI - [Evaluation of bone mineral density in the distal radius of former workers employed at the Aluminum Works]. AB - Fluoride causes an increase in bone mass by stimulation of osteogenetic process. This effect is used in treatment of osteoporosis. Chronic exposure to fluoride in aluminium works can cause an industrial fluorosis, which is characterised by increased mineral content in bone tissue. It is well known that after cessation of fluoride is gradually eliminated from the bone. Probably this same process can exist in patients with osteoporosis after stopping of osteoporosis treatment, so we decided to estimate bone mineral density in former workers of aluminium works. For investigation a group of 169 of men in mean age of 50.0 years, all of whom had worked for at least five years (average 12.9 yrs) in Skawina aluminium works before their closure in 1981, was selected. The control group was 29 men in the same age not exposed for fluoride. In all patient bone densitometry in distal and ultradistal region were evaluated. Decreased bone mineral density was found in workers of aluminium workers, compared to the control group, particularly in age groups of 40-44 and 50-54 years. Differences were bigger in measurements of trabecular bone. PMID- 9333881 TI - [Evaluation of mineral density in the distal radius during the course of rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) of progressive systemic disease predisposing for osteoporosis. Inflammatory process, applied treatment as well as considerably impared efficiency of motor organ create conditions for osteoporosis. The changes of bone mineral density in RA were assessed in 50 patients treated for various form of RA, at the Rheumatological and Rehabilitation Hospital in Cracow. The age of patients ranged from 21-79 yrs: the mean age was 50 years. The group consisted of 46 (92%) females and 4 (8%) males. Apart from standard clinical examinations there was assessed in all cases mineral density in distal radius using Osteometer DTX 100. Mineral density BMD was estimated in distal and ultradistal region of radius. All patients were qualified into 4 groups depending on the stage of radiological changes according to Steinbrocker. Group I included 16%, group II 30%, group III-30%, and IV-24% of patients. Steroid therapy was applied in 20 (40%) cases. The results showed progressive decrease of mineral density BMD in distal radius in patients with advanced RA. It was also observed that in RA patients mineral density defect occurs earlier in trabecular than in cortical bone. PMID- 9333882 TI - [Osteoporosis during development--selected problems]. AB - In children and the youth it is secondary osteoporosis (OP) rather than idiopathic one which occurs more often; its multidirectional pathogenesis is usually ascertainable. Secondary OP, mostly generalized, is diagnosed in the course of such hormonal disturbances as: primary hyperparathyroidism, hyperthyroidism, hyperadrenalocorticalism, hyperpituitarism (with excess of growth hormone) and in hypogonadism. Another group of diseases implicating OP are connective tissue pathologies: congenital (osteogenesis imperfecta, collagenopathies) and acquired (Juvenile chronic arthritis). A serious problem for a pediatrician is the iatrogenic OP resulting from a long-term use of some medicines (glucocorticosteroids, hydantoin derivatives, barbiturates), or long lasting immobilization for surgical and orthopaedic reasons, or from chronic general diseases. Osteoporosis accompanying pathological states of the skeletal and nervous systems (with paralyses and pareses) is particularly intensive and difficult for treatment. Osteoporosis in developmental age may cause disturbances in natural development of the skeleton, which leads to deformities in the skeletal system and to the formation of faulty postures. Lower body height is a frequent complication resulting from OP in children and the youth. PMID- 9333883 TI - [Prevention and treatment during the postmenopausal period]. AB - Osteoporosis will always be a subject of human interest. Thus, it has to be governed by general laws, which-in medicine-still too often differ from the latest truths of nature, to pamper to some fashionable medical views. Its diagnosis makes the doctor feel good, because it explains the cause of the symptoms or disorders. The process of osteoporosis clearly intensifies in the period of climax and therefore, not only the menopause but also the symptoms of climacteric syndrome, anovulatory cycles after 35 years of life, past hemorrhages connected with pregnancy, infections of mammary glands in puerperium involving lack or shortening down to 3 months of natural breast-feeding, states after treatment of sterility, and particularly the infertility, as well as after excessively prolonged use of oral contraception, are indications for diagnostic examination. The aforesaid limit of 35 years of age allows the commencement of prophylactic hormonal prevention of postmenopausal disorders and diseases a lot earlier (more than ten years), instead of their belated-literally substitutive treatment. The most appropriate are physiological doses, which-by supporting the endogenic natural source-treat without post-pharmacological side-effects, of which the most frequent are unwanted postmenopausal bleeding and/or hemorrhages from the genital tract. The principal rule of postmenopausal hormonal treatment is to adjust not only to the doctors orders, but also to one's own feeling and the estimated effectiveness of currently administered doses. PMID- 9333884 TI - [Rehabilitation in osteoporosis]. AB - The paper discusses some of the factors that affect the onset of osteoporosis, and provides an original model for rehabilitation procedure in this disorder. The type of motor exercises used are indicated, and the sequence in which they are done. There is also a discussion of the significance of the application of physiotherapy equipment. Attention is also drawn in the paper to the possibility of the patient applying rehabilitation procedures independently, under only periodic supervision by the physician. PMID- 9333885 TI - [Drug abuse in 1997 in Poland]. PMID- 9333886 TI - [The future of clinical toxicology in Poland]. PMID- 9333887 TI - [Evaluation of the treat of acute poisoning with chemical compounds among adult inhabitants of Krakow in 1995]. AB - The goal of this study is to evaluate the number and the reasons (poison and structure) of acute poisonings which occurred among Krakow adult inhabitants in the year 1995. Under analysis there were 3003 people treated at the Department of Clinical Toxicology and 210 poisoned who died at the place of accident prior to any treatment was conducted. The group of hospitalized persons consisted of 63.7% men and 36.3% women, and the group of people who died at the place of accident consisted of 89.5% men and of 10.5% women. The overall coefficient of poisonings during the year 1995 was 48.3; for men was 66.7 and 32.5 for women. A drugs (45.1%) followed by ethanol (42.8%) were the most common cause of acute poisonings. The mortality rate of the treated patients was low (0.7%), but while including those people who died at the place of accident prior to any treatment, the mortality rate rose up to 7.2%. That increase in the mortality rate was caused mainly by fatal cases due to ethanol and carbon monoxide poisonings. PMID- 9333888 TI - [Acute poisoning with psychoactive substances among adult abusers of the Krakow population]. AB - The aim of the study was to evaluate a frequency and the course of acute poisonings with psychoactive substances in abusers, considering the kind of toxic substance. 25% of all acute poisonings treated in the Department of Clinical Toxicology in the year 1996 that were acute poisonings with psychoactive substances among the abusers of Krakow adult population. Alcohol followed by narcotics and drugs and by organic solvents was predominant cause of acute poisonings in that group. Opiates derivatives as the so called kompot produced at home from poppy straw or juice of poppy head were the most common narcotics. Pure amphetamine was detected only in 5.3%, while as combination with other narcotics (opioids, THC, LSD) was detected in 31.2% of narcomans and drug abusers. The combination of opioids, barbiturates and benzodiazepines was predominant cause of acute intoxications with mixed substances abuse. That was followed by combination of opiates and barbiturates in the group of patient identified positively as two or more substance abuse in the body fluids. Tramal and benzodiazepines were the most detected drugs responsible for acute intoxications between drug abusers. Acute intoxicated ethanol abusers aged from 30-49 years and were the oldest group compared to narcotics and drug abusers (20-29 years) and to organic solvent dependent patient (15-19 years). PMID- 9333889 TI - [Poisoning with narcotics and hallucinogens in material from the Clinic of Acute Poisoning in Lodz (1993-1996)]. AB - Our material consists of the patients treated for poisonings with street-drugs in Clinic of Acute Poisonings, Lodz, Poland during the period 1993-1996. During the time of 4 years, the number of hospitalised patients have grown up 4.5 times. The structure of used drugs has also changed. Up to 1993 the most popular were opiates derivatives and narcotic analgetics-morphine derivatives. In 1996 almost all narcotics are common, opiates (both natural and synthetic), cocaine, LSD, amphetamine and commonly used (by youngsters especially) Cannabis sativa (marihuana etc). The most common sources of poisoning in our material were opiates and amphetamine, followed by tetrahydrocannabinol derivatives-marijuana. PMID- 9333890 TI - [Selected medico-legal problems in cases of poisoning with substances of abuse]. AB - In toxicological examinations of substances of abuse with respect to medico-legal purposes many problems have been discussed. A selection of biological material and proper analytical methods in confrontation with interpretation of results in reference to the subject in question seems to be a central toxicological issue. Blood and urine samples are usually used in cases of human. The problem is more complicated if it concerns dead subjects on the account of postmortem processes in a body. From this point of view, in fatal cases of poisonings experience proves, besides blood and urine also other body fluids such as: cerebrospinal fluid, vitreous humor, perilymph and bile turned out to be usefull for examinations of various xenobiotics. The analytical methods (FPIA, HPLC, GC-MS) have been used for qualitative and quantitative evaluation in wide range of xenobiotic concentration in biological matrix. Interpretation of the results including such problems as toxic interaction, metabolism, data base has been discussed in the study on the basis of intoxication cases with substances of abuse. PMID- 9333891 TI - [Correlation of carbamazepine levels in blood with clinical poisoning states, evaluated with the help of the APACHE II system and the Matthew coma scale]. AB - Acute carbamazepine (CBZ) poisoning occurs recently quite often. Symptomatology of poisoning is variable but usually various degrees of consciousness impairment prevail. 77 patients (36 women and 41 man, mean age 31.5) were hospitalized last two years in this Centre. Clinical condition was evaluated in a regular descriptive way, classifying the degree of coma according to the Matthew scale but also by calculating the scores according to APACHE II and TOXSCORE. Serum CBZ was measured. A slight falling trend was found of the relation of the serum CBZ in the range 6-37.8 micrograms/ml (38 micrograms/ml = median) to APACHE II score and the TOXSCORE and slight rising trend of the relation of the serum CBZ to the degree of coma. The significance of this trend rose essentially at the serum CBZ levels of more than the median 38 micrograms/ml. The causes of the non significant correlation of serum CBZ (in particularly range) to the clinical condition is discussed. The individuals factors of the patient and possible effect of other, unknown drugs taken together with CBZ seem to play a minor role at the concentrations above 38 micrograms/ml. A very precise correlation has been found at the serum concentrations exceeding 40 micrograms/ml. PMID- 9333892 TI - [Suicidal poisoning with benzodiazepines]. AB - In the period from 1987 to 1996, 103 patients with suicidal benzodiazepines poisoning were treated, including 62 women and 41 men from 16 to 79 (mean 34) years old. 23 persons were poisoned only by benzodiazepines, in 80 remaining cases intoxications were mixed eg. including benzodiazepines and alcohol, tricyclic antidepressants, barbiturates, opioids, phenothiazines. The main causes of suicides were mainly depression, drug addiction and alcoholism. Nobody died in the benzodiazepines group, while mortality rate in the group of mixed poisoning was 4%. Prescribing benzodiazepines by physicians was quite often not justified and facilitated, among others, accumulation of the dose sufficient for suicide attempt. Flumazenil was efficient for leading out from coma in 86% of cases with poisoning only by benzodiazepines and 13% of cases with mixed intoxications mainly containing benzodiazepines and alcohol or carbamazepine. PMID- 9333893 TI - [Analysis of observations in the admissions room of the Krakow Clinical Toxicology Department of substance dependent persons in relation to changes in central nervous system metabolism caused by substances of abuse]. AB - Authors presented the course of clinical observation of 393 acutely intoxicated patients dependent on psychoactive substances (28%) or ethanol (72%) who were discharged from the hospital at their own request before diagnosis and treatment were completed. In the first stage of detoxification the different intensity symptoms of toxic coma (25% of patients), behavioral changes-agitation and aggression (16%) and also the disturbances in consciousness accompanied by tremor and vegetative storm (15%) were observed. Authors pointed out that among others the neurobiological changes in brain reward systems of patients who discontinued diagnosis and treatment could be endangering for both the patient and the doctor. They postulate that criteria of life threatening state should by more accurately defined and the methods of full security for patient and doctors should be worked out. PMID- 9333894 TI - [Use of naloxone in acute poisoning with opiates in substance dependent persons]. AB - Acute poisonings with drugs of abuse in Poland in 1995 made up 3-4% of all cases of intoxications. In Krakow the most common cause of drug addiction resulting in life threatening states is the use and abuse of opiates derivatives produced by drug addicted people themselves. This domestic product, so called "kompot" or Polish heroin is produced from poppy straw or juice of poppy head (Papaver somniferum). "Kompot" shows the variable contents of heroin, 6-MAM, 3-MAM, morphine, acetylocodeine and codeine. Papaverine, thebaine and narcotine were also detected. The aim of the study was to investigate the frequency of application and the efficiency of naloxone in the treatment of acute poisonings with "kompot" in the subculture of Krakow drug abusers. 91 patients were treated in the Department of Toxicology in 1996 from the acute poisoning with "kompot". There were 24% of women and 76% of men. The age average was 25.2 years. In 61.5% of cases patients intake only "kompot", in 27.5% "kompot", benzodiazepines and barbiturates, in 6.6% "kompot" and amphetamine and only in 4.4% "kompot" with ethanol. In 10% of poisoned naloxone was applied only at the place of accident. In 58% only in the Department of Toxicology. 32% were treated with naloxone both at the place of accident and in the Department of Toxicology. At the place of accident in most cases the single dose was usually applied (mean dose 0.48 mg) and 76% of patients required the additional treatment with naloxone after the admission (mean dose 1.92 mg). Only 5.5% of acute intoxicated with kompot" drug abusers needed intensive care treatment. Complications in the form of vomiting and excitement were a rare case-5.5%. PMID- 9333895 TI - [Dependence producing substances in the practice of the Insitute of Forensic Experts in Krakow]. AB - Different pieces of evidence connected with drug addiction have been used as material for studies carried out at the institute of Forensic Research in Krakow since the end of the sixties. The number of expert's opinions concerning these questions is constantly on the increase. Work done by the institute in this field has led to the creation of a list of dependence producing substances and their substitutes used by addicts, the methods of illicit production, and the ways and usable forms in which they are introduced into the system. The author describes the most popular drugs and their substitutes used in Poland. The number of these substitutes is big. Among them are: different medicines (e.g. Parkopan, Aviomarin) and solvents (toluene), various vegetable raw materials (e.g. Folium Stramonli, Pallocybe semilanceata). At the same time the use of genuine narcotics like morphine derivatives originating from Papaver somniferum and Cannabis sativa products, obtained by unusual methods of domestic production, was observed. The number of cases involving amphetamine and its analogues increased last year. The presence of opium alkaloids (morphine, codeine, papaverine, thebaine and narcotine as well as acetyl derivatives of morphine (heroine, 3 monoacetylmorphine, 6-monoacetylmorphine) and codeine (acetylcodeine) was detected in "kompot" by means of TLC, HPLC and GC/MS methods. The author also presents, from the analyst's point of view, analytical and interpretational problems of drugs of abuse, and regulations connected with drug addiction and with road traffic offences. PMID- 9333896 TI - [Alcohol and suicide attempts]. AB - A large number of patients (30%) admitted to the Clinic of Acute Poisonings, Lodz, Poland, with suicidal drug poisoning have been found to act under the influence of alcohol. Sociological interview revealed that 6% of them were alcoholics and 3.8% reported to have repeated the suicidal attempt because of alcohol addiction. From the sociological point of view the female and adolescent (15-18 years old) family members are most vulnerable as they are frequently exposed to highly stressful conditions which may eventually result in their becoming addicted to alcohol as well. PMID- 9333897 TI - [Co-dependence--diagnosis and therapy]. AB - The aim of the paper was to present some kind of framework for understanding co dependence as the dysfunction in the close relationship with person, who is addicted to alcohol. The concept of co-dependence outlined here assumes that addiction to alcohol is at least fourfold disease. We are accustomed to thinking of it as having three aspects-physical, emotional (psychological) and spiritual all of which apply directly to the addicted person. But that addiction is also family disease. Everyone around addicted person also suffers. Co-dependence is one of the consequences of that family syndrome. In the paper we present the diagnostic criteria for co-dependence and the current treatment approach. PMID- 9333898 TI - [Specificity of alcoholism and resulting methods of therapeutic practice]. AB - Addiction to alcohol is a bodily disease (DSM IV) kept up by certain psychological mechanisms, which are created in order to preserve and justify the need to drink. In the course of alcoholism those mechanisms are modified to form psychological addiction, thus creating, alongside with physical addiction, twofold psychopshysiological addiction. The specific character of the disease requires both medical treatment and psychotherapeutic actions carried out according to certain definite principles aimed at disarming the defence mechanisms which justify a persons right to drink. The actions of therapists working in order to break off a persons addiction should be of approving and instructive character, and at the same time manifest the therapists awareness of the interdependence of physical as well as psychological aspects of the disease. Due to the chronic character of alcoholism and the resulting threat of relapse, what is of particular importance in maintaining ones sobriety are AA support groups. PMID- 9333899 TI - [The responsibility of the emergency service physician and patient consent to treatment and hospitalization]. AB - Amenability of emergency service physician for the treatment given without patient consent has been presented in the study. Depending on circumstances it can be penal, civil, disciplinary and professional responsibility. The study has been annotated with current legal and ethical rules, which should be not only commonly known to physicians but also respected to avoid legal consequences. PMID- 9333900 TI - [Prevention of narcotic dependence in Polish legal regulations]. AB - This paper presents some aspects of Polish legislation against chemical dependency. The legal regulations and rules related to enforcement and prevention of drug addiction are listed. Also included is the special insight of the Rule for Prevention of Drug Addiction of January 31, 1985. PMID- 9333901 TI - [State and local administration, alcoholism and narcotic addiction]. AB - An important social and medical problem of alcoholism and drug addiction in Poland is presented. Alcoholism is a serious danger for individuals, families and society because it is so common and due to its consequences. Described is a three stage system of preventing measures being undertaken by state and local administration together with local community institutions. By the example of the activities performed by the plenipotentiary of the voivode of Krakow a duties of such institutions are presented. The special role is played by the high medical personnel according to the prevention and early detection of potential threats. PMID- 9333902 TI - [Acute oral poisoning with isopropyl alcohol in alcoholics]. AB - The symptoms and clinical course of repeated poisonings with isopropyl alcohol in ethanol addicted 43-years old man is presented in the study. The blood and urine concentrations of isopropanol and acetone and also the blood ethanol concentration are given. The biological half-life of isopropanol calculated for the first 12 hours from admission to the Clinic was 6.9 hours. The biological half-life of acetone in the first 6 hours of hospitalisation was 5 hours, and then a considerable slowing down up to 29 hours was noted. The clinical symptoms, similar to ethanol poisoning, including strongly manifested catatonia and ketonuria without metabolic acidosis indicate the isopropanol intoxication. Hypokalemia should to be taken into consideration in treatment of isopropanol poisoning. An intravenous bicarbonates should be administered carefully e.g. when rhabdomyolysis occur. PMID- 9333903 TI - [Atropa belladonna poisoning suggesting severe post-traumatic brain damage]. AB - In the paper I have described a case of a student of the University of Technology who has suffered a severe Atropa belladonna intoxication. Originally the student has been sent to the Surgery Department by the ambulance service doctor who had recognized the symptoms of the post-traumatic brain damage. The diagnosis of the post-traumatic brain damage has been excluded during the following diagnostic investigation, the symptoms represented by the patient and his bad general condition, however, has still reminded the same. As the next step the patient has been transferred to the Toxicology Ward according to our suggestion of Atropa belladonna intoxication, where the diagnosis has been confirmed. The patient has regained consciousness after 10 hours of the therapy (with the use of the respirator) and has been discharged from the hospital in a good general condition after 11 days. This case of intoxication has been an example of an experiment done by a very inquiring student on himself. PMID- 9333904 TI - [Narcotic dependence problems studied at the Poison Center in Wroclaw]. AB - Poison Centre in Wroclaw is appointed to the treatment of the most serious cases of poisoning in the Lower Silesia country. Drug addicts merely make 3% of all patients undergoing a treatment. Only few of them show symptoms of severe poisonings with drugs of abuse. Majority of these patients does not require specialised toxicological treatment and nor they agree for detoxification. New chemicals of abuse, like marijuana or LSD, appearing nowadays on Polish illegal market, does not make a toxicological problem in our region yet. PMID- 9333905 TI - [Acute poisoning with narcotics in material from the Ward of Internal Diseases and Acute Poisoning in Tarnow]. PMID- 9333906 TI - [Suicidal poisoning of health service personnel with graduate level education in material from the Acute Poison center in Lublin from 1992-1996]. AB - Suicidal poisoning of postgraduate health service personnel are not described in the literature. We present the experiences on that field of the Acute Poisoning Centre of Jan Bozy Hospital in Lublin. From 1992 till 1996 we have recorded 20 intoxications. Among them the poisonings of the medical doctors are the most frequent. Psychotropic drugs and ethanol were mainly used. One insulin poisoning was mortal. PMID- 9333907 TI - ["Cold turkey" detoxification of drug, narcotic and chemical addicts conducted at the Occupational Diseases and Toxicology Ward of the Polish Center of Occupational Medicine in Poznan in 1986-1996]. AB - In this paper we have discussed the method of the "pure" detoxification (without the use of any psychotropics) of patients addicted to drugs, narcotics and chemicals treated at the Occupational Diseases & Toxicology Ward of the WCMP in Poznan. The material includes 152 cases of addicted patients who were treated in the Ward in the period from 1986 till 1996. Our research proved that the applied method is safe for the patients. We also found that addicts sharing hospital room with general medicine, not addicted patients, experience a positive effects in the process of treatment and resocialization. PMID- 9333908 TI - [Essential, economic and social significance of the Poison Information Center for public health]. AB - The role of toxicological information consisting of chemical substance toxicity, clinical signs and symptoms, treatment and diagnosis of poisonings is presented in the study. These data are collected in computer programmes TOKSY 5, TOKSY7 (MacroSoft Warazawa) which together with American CCIS data base (MICROMEDEX, Denver, Colorado) and IPCS INTOX Database are helpful instrument while giving the toxicological information or/and medical consulting. All cases are registered in IPCS (International Programme on Chemical Safety) INTOX SYSTEM data base that enables epidemiological analysis and also an evaluation of economical and social effects caused by poisonings. PMID- 9333909 TI - [Basic dictionary of terms used as street slang by narcotic addicts]. PMID- 9333911 TI - [Sprue syndrome]. PMID- 9333910 TI - [Whipple's disease: molecular biology--clinical aspects--therapy]. AB - Whipple's disease is a rare systemic bacterial disease, affecting mostly middle aged white men. Its clinical presentation is very variable. Arthralgias, fever of unknown origin or central nervous system symptoms may precede the more typical gastrointestinal manifestations with weight loss and chronic diarrhea. The diagnosis is based on the histopathologic demonstration of PAS-positive macrophages in duodenal or jejunal biopsies. In addition, it has become possible to detect the causative actinobacterium Tropheryma whippelii by amplification of a Whipple-specific PCR product. Therapeutic options are a sequential therapy with penicillin-streptomycin, followed by trimethoprim-sulfamethoxazole (TMP-SMX) or a monotherapy with TMP-SMX. These therapeutic modalities usually eradicate the infection and result in a rapid and sustained improvement of the Whipple-specific signs and symptoms. In view of the frequently uncharacteristic initial clinical presentation (arthralgias, fever, CNS symptoms), it is therefore of utmost importance to consider Whipple's disease in the differential diagnosis of the clinical signs and symptoms. PMID- 9333912 TI - [Intestinal diseases in HIV infection]. AB - The gastrointestinal tract is very frequently affected by the manifestations of the acquired immunodeficiency syndrome (AIDS). A variety of opportunistic viral, fungal, bacterial, protozoal and helmintic infections and different unusual malignancies such as Kaposi's sarcoma, non-Hodgkin's lymphoma and papilloma-virus associated anal cancer are responsible for much of the morbidity and mortality in AIDS. Because specific therapy is not always available, in particular diagnosis of potentially infections should be attempted. PMID- 9333913 TI - [Treatment of stenoses of the gastrointestinal tract]. AB - Flexible endoscopy seems to become the standard method for diagnosis and therapy of stenoses in the gastrointestinal tract. For dilatation of the lumen different methods for bougienage and different types of balloons can be used. For ablation of tissues the Nd:YAG-Laser and conventional high-frequency surgical methods are complemented by the newly developed Argon Plasma Coagulation (APC). The characteristics of APC ideally meet the requirements of an endoscopic tool. The plastic tubes used for bridging stenoses become gradually replaced by the new self-expanding metal prostheses, which have a lower rate of complications and give the patients more quality of life. The problem of tumor ingrowth however ist not yet solved conclusively. Endoscopic methods for decompression can avoid the emergency operation of colonic obstruction. It is important to be aware of the possibility to combine all these methods primarily or secondary to get best results and to minimize risks. To offer an optimal treatment for gastrointestinal stenoses it is necessary to provide the whole armamentum of interventional endoscopy including synchronous fluoroscopy. PMID- 9333914 TI - [New hepatitis viruses]. AB - Until 1987 only the hepatitis viridae A, B and D were discovered. All other viral agencies of hepatitis were summarized under the term non-A-non-B virus. By modern methods of molecular biology and recombinant gene technology, several new hepatitis viridac could be identified and their structure partly characterized. The hepatitis virus E (HEV) is predominantly transferred through the fecal-oral path, the hepatitis viridae C and G predominantly parenteral. The structure and molecular biology of these new hepatitis viridae as well as the pathogenesis, epidemiology, diagnosis, clinical manifestation, prophylaxis and therapy of the various types of hepatitis are described and compared with the wellknown hepatitis viridae A and B. PMID- 9333915 TI - [Extrahepatic manifestations of HBV and HCV infection]. AB - Acute as well as chronic infections by HBV and HCV can be associated with extrahepatic disease. As in liver disease of non-viral etiology, several extrahepatic manifestations can be observed that are non-specific for HBV or HCV, e.g. those clinical signs and symptoms frequently encountered when cirrhosis of the liver is present. In addition, distinct clinical conditions have been described in which the unterlying liver disease is specifically due to HBV or HCV infection. In chronic HBV infection, glomerulonephritis (of the membranous and of the membranoproliferative type) and polyarteritis nodosa have been observed. The pathogenesis of both conditions involves the deposition of circulating immune complexes. Although controlled trials are lacking, interferon therapy appears to be beneficial. The use of immunosuppressive drugs and of plasmapheresis should probably be limited to the initial phase of treatment. In chronic HCV infection, mixed cryoglobulinemia (and ensuing systemic vasculitis) is a frequent extrahepatic manifestation. The clinical presentation might include the presence of purpura, arthralgias, weakness and renal involvement. Nowadays, HCV can be regarded as the etiological agent of a form of mixed cryoglobulinemia formerly considered to be 'essential'. In addition, an association to lymphoma has been postulated recently. Interferon alpha has been used for treatment with similar response rates as those observed in HCV-infected patients without cryoglobulinemia. Interestingly, the antiviral activity of interferon, e.g. normalization of transaminases and loss of serum HCV RNA, was closely related to the beneficial effect on cryoglobulinemia. An association of Sjogren's syndrome and of porphyria cutanea tarda to HCV infection (though claimed before) remains questionable. Serological markers of autoimmunity, e.g. antinuclear antibodies or anti-LKM-1, are present in many patients with chronic HCV infection. The clinical relevance of this latter observation appears low however, particularly since the decision for interferon treatment is hardly influenced by their presence. PMID- 9333916 TI - [Hepatic diseases caused by drugs]. AB - Drug-induced hepatotoxicity covers the clinical and pathological expressions of almost any acute or chronic liver disease. Liver damage is due to intrinsic toxicity of the drug (paracetamol) and/or immunoallergic mechanisms (halothane). Acute injury may be cytotoxic or cholestatic. Cytotoxic injury is characterized by necrosis (paracetamol, halothane) or steatosis (valproate). Cholestatic injury can be associated with immune-mediated portal inflammation (chlorpromazine) or solely attributed to inhibition of transport systems (cyclosporin A). Chronic drug-induced disorders include chronic active hepatitis (methyldopa), autoimmune hepatitis (tienilic acid), alcoholic steatohepatitis-like reactions (amiodarone), indolent fibrosis (methotrexate), chronic cholestatic diseases, vascular lesions, and hepatic neoplasms. The clinical and morphological picture of many drug induced liver diseases is nonspecific. Diagnosis is based on thorough drug history, temporal relationship, time-course of liver dysfunction, and the exclusion of other causes. Treatment consists of instant withdrawal of the suspected drug and administration of acetylcysteine as early as possible in case of paracetamol intoxication. Drug-induced liver diseases are usually reversible, but prolonged treatment leads to progression and liver cirrhosis. PMID- 9333917 TI - [Molecular biology 1996: gastroenterology and hepatology]. AB - Advances in molecular genetics have clearly improved our understanding of the pathogenesis as well as the diagnosis, therapy and prevention of gastrointestinal and liver diseases. The molecular diagnosis of a number of maglignant or potentially malignant gastrointestinal diseases as well as of several infectious liver diseases is on its way into clinical practice. Gene therapy and molecular preventive strategies, by comparison, have been less developed and need further refining before they will become integral part of the patient management. PMID- 9333918 TI - [Burnout and reaction to stress]. AB - The recent flurry of attention to burnout syndrome still leaves numerous questions unanswered. One of them is the relationship between individual factors and the development of burnout. An understanding of the individual factors underlying burnout must include an assessment of the individual reactivity to stress. The occurrence, distribution and relationship with stress reactivity of the three dimensions of the burnout syndrome (emotional exhaustion, depersonalization and lowered feelings of personal accomplishment) were studied among a representative sample of the different professionals involved in the Primary Care Health System. Our results indicate that stress reactivity could be a variable that modulates the experienced psychopathology, suggesting a predisposition that increases the susceptibility to the development of burnout. PMID- 9333919 TI - [Synovial cyst in the lumbar facet joint. A rare cause of lumbar-sciatic pain]. AB - Synovial cysts of the facet joints are uncommon lesions which may be asymptomatic or present as low back pain, with or without radicular symptoms. They are considered to be secondary to trauma or degenerative joint disease, and they occur more frequently in patients with spondylolisthesis. Diagnosis is normally achieved with computed tomography or magnetic resonance, which show a cystic lesion located laterally adjacent to the facet joint. We review the literature and report a patient who presented with first sacral nerve root symptoms attributable to a ganglion cyst of the left L5 S1 facet joint. Treatment was curative and consisted in excision of the cyst. PMID- 9333920 TI - [Bronchiolitis: conceptual review]. AB - Bronchiolitis is a term which brings together several processes which may confuse anyone who is not familiar with the subject. We will review these processes by focusing on the most common types. PMID- 9333921 TI - [What is health?]. PMID- 9333922 TI - [Gynecomastia]. AB - The development of excess breast tissue in the male, known as gynecomastia, is a common finding seen by physicians. It is a source of embarrassment to the adolescent and of surprise and concern to the older male. The main challenge to the physician lies in separating the patients with gynecomastia as a symptom of a serious underlying disease, requiring specific treatment, from the vast majority of individuals with gynecomastia, in whom there is no underlying pathology. Numerous pathogenic mechanisms have been involved in the genesis of gynecomastia, leading all to an increased net estrogen/androgen ratio that effectively acts at the target tissue (breast). Special emphasis must be done in puberal gynecomastia since 50 to 75% of boys in these developments phase have a breast enlargement. In the present paper we provide an overall review of hormonal pathogenesis and diagnostic clues, as well as current therapeutic guidelines regarding gynecomastia. PMID- 9333923 TI - [Meloxicam]. PMID- 9333925 TI - [Surgical treatment of esophageal cancer]. PMID- 9333924 TI - [The obligation of solidarity among physicians. A model from the past]. PMID- 9333926 TI - [Progress of heart surgery for infants]. PMID- 9333927 TI - [Ultrasonic diagnosis and heart surgery]. PMID- 9333928 TI - [Surgical treatment of tricuspid valve insufficiency]. PMID- 9333929 TI - [Learning experience from the "death conference"]. PMID- 9333930 TI - [On limitation on the indication of functional pneumonectomy]. PMID- 9333931 TI - [HIV-associated tuberculosis in Africa exemplified by Zimbabwe]. AB - In Africa, a rapid increase of human immunodeficiency virus (HIV)-associated tuberculosis cases has been observed; 80% of a worldwide 6 million dually infected persons live in this part of the world. The annual risk of progression to clinically overt tuberculosis in dually infected persons approaches the lifetime risk in persons with tuberculosis but no HIV infection. Zimbabwe is an example which illustrates the rapid increase in notified tuberculosis cases since 1985, accounted for primarily by HIV-associated tuberculosis cases. In sputum smear positive HIV-associated tuberculosis, classical symptoms are reported with the same frequency as in HIV negative cases. Thus, case-finding activities need not be altered. In sputum-smear negative patients, reliable diagnostic tests are not available. Therapeutic trials are widely used and this causes overdiagnosis of tuberculosis. Extrapulmonary manifestations are common in HIV-associated tuberculosis. A majority of lymph node enlargements, pleurisy and pericarditis in Africa are now due to tuberculosis. If compliance is ensured, response to chemotherapy is excellent, but overall case fatality and relapse rates are increased. The cost-effectiveness of tuberculosis control programmes using directly observed therapy for at least the first 2 months of treatment is well established. With the prominent global significance of tuberculosis and the possibility of cost-effective interventions, a commitment to the fight against the worldwide epidemic is more important than ever before. PMID- 9333932 TI - [How is hemolytic-uremic syndrome in childhood acquired in Switzerland?]. AB - Intestinal infections with shigatoxin-producing Escherichia coli or Shigella dysenteriae type I play a major role in the pathogenesis of the hemolytic-uremic syndrome in childhood. Escherichia coli has been repeatedly detected in the intestines of healthy cattle. Twenty-seven children with hemolytic-uremic syndrome were treated at our hospital between June 1990 and March 1997. Factors indicating a possible previous contact with bovine intestinal content were found in 18 out of the 27 patients: parents stockbreeders (n = 7), recent visit to a cowshed or contact with cowdung or manure (n = 5), residence in a rural cattle breeding area (n = 5), or consumption of raw milk (n = 1). The factors mentioned were found in 5 out of 27 control patients (p < 0.01). Two children experienced hemolytic-uremic syndrome after a stay respectively in Egypt and Tunisia. Our results indicate an important source for acquisition of hemolytic-uremic syndrome in childhood. Observing simple hygienic rules such as washing of hands and pasteurization of milk is likely to have a positive influence on the incidence of this illness. There are also grounds to consider adding the hemolytic-uremic syndrome to the list of travel-related diseases. PMID- 9333933 TI - [Cost effectiveness of ACE inhibition in therapy of chronic heart failure in Switzerland: evaluation based on the SOLVD study]. AB - BACKGROUND AND OBJECTIVES: Morbidity and mortality data in Switzerland underline the socioeconomic importance of heart failure. In the SOLVD study (Study on Left Ventricular Dysfunction), cardiovascular morbidity and mortality were reduced with the ACE inhibitor enalapril in patients with heart failure. The economic implications of this treatment were analyzed in a retrospective economic analysis from the perspective of Swiss third party payers. PATIENTS AND METHODS: Source of the economic analysis was the SOLVD study data. This prospective study was placebo-controlled, double-blind and had a mean follow-up of 3.45 years (41.4 months), involving 2569 patients with heart failure, mainly in NYHA classes II and III. Costing data for treatment with enalapril, the per diem charges for hospitalization and the average length of hospital stay were retrieved from published national sources. The costs of in- and output were calculated and compared for the two treatment groups in a cost-efficacy analysis. RESULTS: Additional treatment with enalapril resulted in an additional cost of 2.5 million Swiss francs. These incremental costs were, however, offset by reduced hospital costs (CHF 6.45 million savings) in the enalapril group. For the complete treatment cohort of the SOLVD study, the net savings were approximately 4.26 million Swiss francs. CONCLUSIONS: From the clinical point of view, treatment with ACE inhibitors leads to a reduction in the progression of heart failure and reduced cardiovascular morbidity and mortality. With respect to health economics, it can be demonstrated that treatment with enalapril does not only offer clinical benefits, but that these also translate into impressive economic savings of CHF 3315 per patient. PMID- 9333934 TI - [Reversible posterior leukoencephalopathy: significance in everyday clinical practice]. AB - Reversible posterior leukoencephalopathy (PLE) is a newly recognized syndrome with characteristic radiologic findings. The clinical picture resembles that of hypertensive encephalopathy. PLE is caused by uncontrolled hypertension as well as other neurotoxic conditions. We report on 3 patients with this syndrome. One patient also had transient hydrocephalus which may have been caused by PLE. Clinicians must be aware of this syndrome as its recognition obviates unnecessary diagnostic procedures. PLE is reversible by lowering elevated blood pressure and treating neurotoxic conditions such as uremia. PMID- 9333935 TI - [The "pinch off" syndrome: a complication of implantable catheter systems in the subclavian vein]. AB - We report the case of a 63-year-old male patient who received a subcutaneous Port a-cath system in the right subclavian vein for administration of chemotherapy. Five weeks after successful use, flushing with NaCl 0.9% caused a painless subcutaneous swelling. Fluoroscopy confirmed a leakage below the right clavicle. The catheter was explanted and replaced by a new catheter via Seldinger technique. Two weeks later another leakage occurred and the catheter was removed definitively. Defective equipment was ruled out by the catheter producing company. The "pinch off" syndrome, a rare phenomenon of catheter compression and consecutive catheter fracture, is described, together with the diagnostic signs, the incidence and preventive measures suggested in the literature. PMID- 9333936 TI - [Thoracoscopic splanchniectomy for pain management in inoperable pancreatic carcinoma]. PMID- 9333937 TI - [Diagnostic process in internal medicine: decision analysis or intuition?]. AB - The diagnostic process is a hypothesis-testing approach under conditions of uncertainty. Experience-based intuition plays an important role in complex clinical situations. Serious cognitive errors may be avoidable if possible heuristic problems are considered. Probabilistic reasoning according to Bayes' theorem, especially for determination of pretest probability, is crucial for coping with uncertainty and the myriad available diagnostic tests and procedures. Their proper interpretation depends on fundamental test variables such as sensitivity, specificity and likelihood ratio. Computers and neural networks cannot replace cognitive reasoning and probabilistic strategies in daily clinical practice. Decision analysis is pivotal, particularly when the aim is to maintain high-quality medical care at a time of limited resources. PMID- 9333938 TI - [PFAPA syndrome: periodic fever, adenitis, pharyngitis and aphthous stomatitis]. AB - A syndrome involving periodic fever, pharyngitis, adenitis and aphthous stomatitis is described in 8 children. Attacks are characterized by abrupt onset of fever and, in addition to the above symptoms, by malaise, headache and abdominal pain. Mild leukocytosis and elevation of the erythrocyte sedimentation rate are found in the laboratory. Patients exhibit normal growth and development and are otherwise healthy. PFAPA is clinically benign, with no long-term sequelae. Recognition and diagnosis of the syndrome eliminate the need for intensive work-up. The cause remains unknown. No evidence linking bacterial, viral, or fungal pathogens to this syndrome has been found. No patient has exhibited atypical lymphocytosis or neutropenia, and all patients had normal levels of immunoglobulin. All had received antibiotics early in the course of their illness but without effect. Cimetidine has been discussed in the literature as a possible treatment, but the results are controversial. PMID- 9333939 TI - [Thrombolysis in acute myocardial infarct in everyday clinical practice]. AB - We prospectively included in a database all thrombolyzed acute transmural myocardial infarction patients admitted to our hospital from November 1986 to September 1995. Six hundred and twenty-seven patients (497 males) with a mean age of 61 +/- 12 years (range 26-88 years) were included. 87% were having their first acute myocardial infarction. Different thrombolytic regimens were applied in the emergency room but the vast majority (92%) received t-PA. The median delay between the onset of pain and admission was 2 h 0 min (10 min-22 h). The median admission to treatment time was 40 min (5 min-6 h 20 min). The latter has been shortened (median 55 min from 1986 to 1989 versus 35 min from 1990 to 1995, p < 0.05) during the study period. The rate of intracerebral hemorrhage was 2.4% (confidence interval 1.1-3.5%) and no significant predictor could be found, although patients with cerebral bleeding tended to be slightly older (66 +/- 9 years vs 61 +/- 13 years, p = ns). The rate of false diagnosis was only 4.6%, even when patients with a final diagnosis of unstable angina and/or aborted acute myocardial infarction were included. The in-hospital mortality was 8.8%, a rate similar to those reported in the literature. Using multivariate analysis, negative prognostic factors were higher age (p < 0.001), advanced Killip class at admission (p < 0.001) and elevated peak CPK levels (p < 0.001). These results confirm that thrombolysis for acute myocardial infarction in the emergency room can be done with a short admission-to-treatment time and with an acceptably low rate of false diagnosis. However, our intracerebral hemorrhage rate was clearly higher than generally reported in the literature and may be explained by a different patient selection from that in large randomized studies. PMID- 9333940 TI - [Disseminated mycobacteriosis with M. kansasii in a case of AIDS without HIV infection]. AB - M. kansasii is an environmental Mycobacterium that causes disease mainly in patients with local or systemic immunodeficiency. We present a case with disseminated M. kansasii infection which led to a severe febrile illness lasting for 9 months. The patient had a combined acquired immunodeficiency syndrome (AIDS) which included CD4+ and CD8+ T-lymphocytopenia, monocytopenia and a deficiency in the IgG subclasses 2 and 4. HIV testing was repeatedly negative. In the published literature only 4 of 10 similar patients (this patient included) survived a disseminated M. kansasii infection. Eleven years after the disseminated infection with M. kansasii a second opportunistic infection, disseminated mollusca contagiosa and hyperparathyroidism, developed, and 14 years later the immunologic abnormalities persist. PMID- 9333941 TI - Fear of hope. PMID- 9333942 TI - South wants place at table in new collaborative effort. PMID- 9333943 TI - NIH case ends with mysteries unsolved. PMID- 9333944 TI - Gene mutation provides more meat on the hoof. PMID- 9333945 TI - HIV suppressed long after treatment. PMID- 9333946 TI - Why can't a computer be more like a brain? PMID- 9333947 TI - A subtler silicon cell for neural networks. PMID- 9333948 TI - Tying it all together: epigenetics, genetics, cell cycle, and cancer. PMID- 9333950 TI - [Pathological fractures--rare, sensitive, exciting]. PMID- 9333949 TI - Lymphocyte survival: a red queen hypothesis. PMID- 9333951 TI - [Pathological fractures]. AB - Pathological fractures are defined as fractures that occur in patients with weakened bone due to systemic and local diseases. If osteoporosis is excluded, the frequency amounts to 5% of all fractures treated. The main diagnostic aim is to observe the pathological nature of the fracture and to perform interdisciplinary, causal therapy. For this, diagnostic and therapeutic algorithms are demonstrated, arranged according to osteopathies, constitution anomalies, myelogenous and inflammatory bone diseases, primary and secondary bone tumors and posttraumatic/postoperative disorders. These algorithms should help the trauma and orthopedic surgeon to select the proper conservative and operative therapy and to use all resection and reconstruction possibilities, such as osteosynthesis, alloarthroplasty and bone transplantation. PMID- 9333952 TI - [Scapholunar ligament injuries in acute wrist trauma. Arthroscopic diagnosis and minimally invasive surgery]. AB - The final result of the treatment of distal intraarticular radius fractures depends both on the accuracy of the fracture reduction and on the presence or absence of additional carpal injuries. In particular, lesions of the intrinsic ligaments usually cause severe degenerative damage of the wrist joint if they are missed primarily. With the introduction of wrist arthroscopy these tears can be evaluated and treated earlier. Since 1993 arthroscopically assisted treatment has been performed in 23 patients with distal intraarticular fractures of the radius (mainly C fractures according to the AO classification system or group VII and VIII fractures according to Frykman). SL tears have been found in 11 patients (47.8%), 7 of whom showed marked instability and were stabilised at the time of surgery. PMID- 9333953 TI - [Intramedullary pressure reduction in the femur shaft in total hip endoprosthesis. Effect on the rate of thrombosis]. AB - The incidence of deep-vein-thrombosis is claimed to be reduced by avoiding intramedullary compression in the femur during implantation of the femoral part of a hip prosthesis. A double-blind randomized trial investigated 101 patients with and 102 without avoidance of intramedullary compression. There was no evidence of the benefit of this technique, for the reduction of postoperative deep-vein thrombosis. PMID- 9333954 TI - [Proprioceptive deficits of the knee joint after rupture of the medial meniscus]. AB - We assessed knee joint proprioception in 23 patients with an isolated lesion of the medial meniscus. Thirteen patients were tested prior to their arthroscopic operation, and 10 patients were examined after partial arthroscopic resection of the injured meniscus. As a control group we evaluated 30 healthy volunteers with clinically inconspicious knee joints. For documentation of the proprioceptive capabilities we performed an angle reproduction test. Additionally the subjects were tested with and without an elastic knee bandage, to test the influence of such a bandage on knee joint proprioception. Our results showed that preoperatively proprioception was significantly less good than in the control group. We found no influence of the knee bandage on the proprioception of the injured knee. The postoperative group of patients showed significantly better proprioceptive capability than the preoperative patients. The postoperative results did not differ significantly from those of the control group. PMID- 9333955 TI - [Radiologic and histologic observations in central talus fractures]. AB - In a prospective study of central fractures of the talus, radiological data (plain radiography, CT, MRI) were scrutinized in terms of correlation with histological findings (biopsy taken at time of screw removal). The study looked at nine fractures in eight patients aged 14-48 years. There was one fracture of Hawkins type I, one fracture of type II, four fractures of type III, and three corpus fractures. All were operated on within 24 h after injury; seven had screw fixation, two had adaptation with K wires. The patients were kept non-weight bearing for 14-53 weeks. Duration of follow-up was 2 years or more in all but one patient. In the final radiograph the talus was intact in four patients, while four showed a partial avascular collapse; in one patient, a partial collapse was doubtful. Histology taken 16-52 weeks after injury showed osteocyte-free original bone in all cases, with deposition of new bone varying in extent. Hawkins sign was partially positive in six patients, positive in one, and negative in one. Decalcification as well as fracture healing can be better followed by CT than by plain radiography. MRI appeared normal in five patients, partially pathological in two, and abnormal in one. A positive Hawkins sign and normal signal behavior can be seen as prognostically favorable signs. However, in the case of a central talus fracture, even with a favorable course osteonecrosis of at least a part of the talus with gradual replacement by new bone occurred. Vascularization of the central talus seems to be disturbed more frequently than is reflected radiologically. PMID- 9333956 TI - [Rate of infection in trauma surgery. An overview based on recent German language literature]. AB - Infection rates are important markers for clinical quality assurance. For internal control, they may only be used under the condition of homogeneous data collection and evaluation according to identical standard operating procedures during the entire investigation period. For inter-hospital comparison, they may only be used if additionally the observed patient groups are well defined and comparable. A survey of the infection rates published during the last 6 years in the German traumatological literature (n = 71) indeed shows (concerning series later than 1985) similar infection rates for procedures in less and in more problematic anatomical regions and in clean and contaminated situations of about 2-3%, after open injuries sporadically max. 10%. Finally, it is demonstrated that conclusions concerning a general "risk of infection" based on infection rates for specific surgical procedures are not possible and vice versa. We strongly recommend the future application of a standardized definition of wound infection. The differentiation between deep and superficial infection should be abandoned. For all mentioned "infection rates" it should be indicated whether it is with reference to the risk of infection of a specific procedure or only a general statement. PMID- 9333957 TI - [Experiences with the pulse oximeter during preclinical cardiopulmonary resuscitation]. AB - The use of pulse oximetry during pre-hospital cardiopulmonary resuscitation was prospectively investigated in 23 cases. The question was whether the measurement of oxygen saturation during pre-clinical resuscitation is useful and what conclusions are possible. We were able to obtain confidential data for oxygen saturation in 82.6% (n = 19) of cases. Our results show that pulse oximetry could be used as a means of quality control during resuscitation. It is a useful part of pre-hospital monitoring and should be used during resuscitation as early as possible. PMID- 9333958 TI - [Shock room diagnosis in polytrauma. Value of thoracic CT]. AB - OBJECTIVE: The aim of this prospective study was to evaluate whether early thoracic computed tomography (TCT) is superior to routine chest X-ray (CXR) in the diagnostic work-up of blunt thoracic trauma and whether the additional information obtained influences subsequent decisions on therapy in the early management of severely injured patients. PATIENTS AND METHODS: In a prospective study of 103 consecutive patients with clinical or radiological signs of chest trauma (94 multiply injured patients with chest trauma, 9 patients with isolated chest trauma) who had an average ISS of 30 and an average AIS thorax of 3, initial CXR and TCT were compared after the first assessment in our emergency department (a level I trauma center). Mortality in this group was 10% (n = 10). RESULTS: In 67 patients (65%) TCT revealed major complications of chest trauma that had been missed on CXR: lung contusion (n = 33), pneumothorax (n = 27), residual pneumothorax after chest tube placement (n = 7), hemothorax ((n = 21), displaced chest tube (n = 5), diaphragmatic rupture (n = 2), myocardial rupture (n = 1); in 11 patients only minor additional pathologic findings (dystelectasis, small pleural effusion) were visualized on TCT; and in 14 patients CXR and TCT showed identical pathologic results. In 11 patients neither CXR nor TCT revealed pathologic findings. The TCT scan was significantly more effective than routine CXR in detecting lung contusions (P < 0.001), pneumothorax (P < 0.005) and hemothorax (P < 0.05). In 42 patients (41%) the additional TCT findings did affect, the therapy selected: chest tube placement or chest tube correction in mostly anteriorly located pneumothoraces or large hemothoraces (n = 31), influence on ventilation mode and respiratory care (n = 14), influence on the management of fracture stabilization (n = 12), laparotomy in cases of diaphragmatic lacerations (n = 2), bronchoscopy for atelectasis (n = 2), exclusion of aortic rupture (n = 2), endotracheal intubation (n = 1), pericardiocentesis (n = 1). CONCLUSIONS: TCT is highly sensitive in detecting thoracic injuries after blunt chest trauma and is superior to routine CXR in visualizing lung contusions and pneumo- and hemothorax. Early TCT influences therapeutic management in a considerable subset of patients. We therefore recommend TCT in the primary diagnostic work-up of multiple injured patients with suspected chest trauma, because early and accurate diagnosis of all thoracic injuries along with acceptance of the implications for therapy may reduce complications and improve the outcome in polytraumatized patients with blunt chest trauma. PMID- 9333959 TI - [Follow-up and prognosis of severe accidental trauma in the aged]. AB - Multiple injuries in elderly patients are still a common problem. The present study was performed to investigate mortality and complications in multiple trauma patients aged 65 years or more. A total of 1154 multiple trauma patients with an injury severity score (ISS) of at least 18 points were divided in two age groups: Y: 16-64 years, n = 1022; O: 65-94 years, n = 132. Older patients were injured as pedestrians in most cases (69%), while younger patients were more frequently injured as car and drivers passengers (41%). ISS was comparable in both groups (Y 28 +/- 1, O 27 +/- 1). During ICU-therapy incidence of ARDS (Y 10%, O 11%), multiple organ dysfunction syndrome (MOF; Y 6%, O 9%) and pneumonia (Y 17%, O 21%) were comparable. In contrast, septic complications were more frequent in older patients (Y 19%, O 27%). Length of ICU stay (Y 19 +/- 2, O 18 +/- 1) and ventilation time (Y 14 +/- 2, O 17 +/- 1) were comparable. Mortality was significantly higher in older patients (Y 15%, O 53%). The major cause of death was sepsis in older patients (Y 15%, O 31%) and MOF in younger patients (Y 54%, O 29%). In conclusion, older trauma patients had a higher mortality due to the development of septical complications. PMID- 9333960 TI - [Injuries of the pelvic ring]. PMID- 9333961 TI - [Homologous talus replacement after talectomy in infection and septic talus necrosis. Experiences with 3 cases]. AB - Severe infections of the talus are often associated with complete septic collapse of the talus. In this connection, open fractures with defects or significant comminution have a bad prognosis is as far as reconstruction of the talus is concerned. In the Department of Traumatology, Braunschweig, in 1995 three patients (all male, average age 35.3 +/- 10.2 years) were treated with cancellous bone grafting after talectomy performed because of infection and complete septic collapse of the talus. In two of these cases third-degree open total dislocation of the talus had been sustained. The third patient came to us after undergoing arthroscopy of the ankle region in another hospital. In each case a fulminating infection was the outcome. Following a step-by-step algorithm, in a first step urgent radical debridement with talectomy was done. To maintain approximation between the tibia and calcaneus on one side and the os naviculare on the other, the bony defect was filled with PMMA chains and the external fixateur technique was used for immobilization during treatment of the infection. After second- and third-look procedures a free flap was grown for soft tissue coverage within the first 10 days. After 17.6 +/- 3.3 days the talus was replaced with a cancellous bone graft, combined with double arthrodesis in two cases, and external fixation for the next 4-5 weeks. In the third patient a triple arthrodesis was done. At follow-up after an average of 12 months (range 8-17 months), the bone graft with arthrodesis had been completely integrated in all cases. All patients are free of symptoms in normal life. In the case of severe open fractures of the talus with significant comminution combined with infection and septic bone collapse conservation of the talus is often impossible. The combination of homologous cancellous bone grafting and arthrodesis after talectomy is therefore a good method of keeping any decrease in the function of the foot to a minimum. PMID- 9333963 TI - [Back problems--prevention and therapy. Interview by Christiane Weseloh]. PMID- 9333964 TI - [Muscle strains: recommendations for the athlete]. PMID- 9333962 TI - [Traumatic axial dislocation injury of the carpus and metacarpus with triquetrum fracture. Rare entity or typical injury]. AB - Although there are only 39 reports of compression injuries with axial dislocation of the carpus and metacarpus, there are similar patterns of injury. In all cases the dislocation is located either between the first and second or the third and fourth metacarpals and the corresponding carpal bones so that weak points of the carpal arch can be assumed in these regions. Clinical and radiological findings in a 22-year-old patient with dislocation of the third and fourth metacarpals and a fracture of the os triquetrum of the right hand after compression injury of both forearms with severe soft tissue injuries are described. Wound debridement and subsequent skin grafting were performed. The osseous lesions were reduced and held by two Kirschner wires and an external fixator. Results at follow-up were satisfactory. PMID- 9333965 TI - [Femoropatellar pain syndrome in extensive flexor-extensor malfunction. Case report of a 23-year-old patient]. PMID- 9333967 TI - [The debate concerning hypertrophy or hyperplasia of muscle fibers]. PMID- 9333966 TI - [Mobilizing joint drainage. A new therapeutic concept in treatment of pathologic edema in the postoperative and post-traumatic field]. PMID- 9333968 TI - [Modification of reactivity by rhythmic neuromuscular stimulation]. AB - The rhythmical neuromuscular stimulation (RNS) by Nasarov is a new method for optimizing performance. By transferring mechanical vibration to the tendomuscular system better coordination of peripheral and central nervous system could be achieved. A study with twelve healthy students of physical education dealt with the question of the effects of RNS on drop jumps. After 12 minutes application of RNS the performance drastically decreased. Jumping height lowered and ground contact time increased. Recordings of EMG revealed corresponding alterations in muscle activity, such occurring in overload situations. Plastic deformation of tendon collagen and neuromuscular adaptation as stiffness-reduction of gamma modulation is discussed. PMID- 9333969 TI - [Inline skating--typical injuries and prevention]. AB - In a field study we evaluated 500 inline skaters and documented the risk for injury as well as active and passive prophylactic methods. Inline-skating carries high risk for injuries. A total of 41% of the 500 examined subjects got injured. 73% of the injuries attacked soft tissue, 22% of the subjects suffered in distorsions and 5% fractures. 24% of the 222 injured people required specific therapy. Most of the injuries occur at the upper extremities. The fingers, wrist or elbow were involved in 78% of the fractures and 58% of the distorsions. Only every second skater has skating skills that allow him to break appropriately. 19% wore all necessary protectors. 21% did not use any protecting device. Wrist protectors were used most followed by knee and elbow protectors. PMID- 9333971 TI - [Pronation angle of the rear foot during running in relation to load]. AB - In 20 volunteers the relationship between rear-foot pronation and increasing physical exertion during treadmill ergometry was examined. In order to assess the influence of regularly performed running training a group of 10 endurance trained middle- and long-distance runners (age: 27.4 +/- 4.9 years; weight: 71.0 +/- 8.8 kg; height: 184.2 +/- 8.3 cm) was compared to another group of 10 untrained subjects (age: 24.7 +/- 2.1 years; weight: 73.3 +/- 9.8 kg; height: 179.1 +/- 8.3 cm). The examinations were carried out on a treadmill using a high-frequency motion analyzing system. Heart rate, blood lactate as well as rear-foot pronation were measured. Regarding heart rate and lactate concentration there were significant differences between trained and untrained volunteers. The pronation angle increased with higher speed up to a maximum of 6.54 +/- 4.22 degree for the trained group and 6.84 +/- 4.59 degree for the untrained group. With reference to maximal as well as submaximal stages the pronation angles showed no significant differences between both groups. Following the maximal exercise level the runners performed an additional 3 min run with a velocity reduced by 8 km/h compared to the maximal speed. At this level the total group as well as the untrained group showed significantly greater pronation angles compared to those of the corresponding velocity at the beginning of the test. The extent of the differences, however, was not significantly correlated with the lactate levels. Our results demonstrate that the increase of the pronation angle is a function of the running speed. But there is also an influence of fatigue, which depends neither on the running velocity nor on the lactate levels during exercise. Therefore, further investigations should emphasize the question which factors are responsible for this effect. PMID- 9333970 TI - [Injuries due to inline skating]. AB - In order to elucidate the patterns of injuries associated with in-line skating all patients with a inlineskate injury have been sampled prospectively during summer 1996. 58 patients were included in the study, aged 8 to 54 years (mean 22.2 years). The total number of injuries was 63, that is 1.1 injury in every injured skater. The upper extremity was the region most commonly injured (63.5%), with the distal radius fracture being the most common single fracture (25.4%). The most severe injuries, however, could be found in the lower extremity, including two femoral neck fractures and one pertrochanteric fracture in three patients aged more than 35 years. 22.4% of patients required hospitalization up to 54 days, and 36.2% of injuries had to be treated operatively. The most common single procedure was closed reduction and percutaneous wire fixation of displaced radius fractures. It is concluded that in-line skating imposes a risk of severe injuries especially on first-time skaters aged 35 years and up. The typical skating injury is the fracture of the wrist, a fact pointing out the necessity of the use of appropriate safety gear. PMID- 9333972 TI - [Injuries during handball. A comparative, retrospective study between regional and upper league teams]. AB - To evaluate the injuries in Team Handball 186 male players out of 16 teams were questioned retrospectively at the end of a season. An important difference between practice and competition injuries could be found with an injury rate of 0.8 injuries per 1000 training hours and 13.5 injuries per 1000 playing hours. Injuries were predominant at the lower extremities with ankle injuries being the most frequent injury type. 2/3 of the playing injuries occurred in offense. 1/3 of these offense injuries could be attributed to a counter-attack. These injuries were generally more severe which lead us to a proposition of changing the rules of the game in foul plays in this situation. Elbow injuries in goalkeepers and shoulder injuries in throwers were specific for these types of players. Prophylactic braces were used by nearly 2/3 of the players. PMID- 9333973 TI - [Orthopedic hippotherapy in postoperative rehabilitation of lumbar intervertebral disk patients. A prospective, randomized therapy study]. AB - In a prospectively randomized therapy study the influence of a new therapeutical approach, called "Orthopedic Horseback-Riding-Therapy (OHRT)", was evaluated on the postoperative rehabilitation after lumbal discectomie in 16 patients against an identical numbered control group. In comparison with the reference group the utilization of OHRT not only produced an improvement in the patients' self evaluation of their postoperative condition (McNab Score). Also a significant reduction of postoperative work disablement could be achieved. Compared with the reference group influences of previously detected negative psychic predictors (Hs and Hy scales of MMPI) could be reduced. Thus the OHRT is a serious therapy concept in postoperative treatment of patients with lumbal disc herniation. PMID- 9333974 TI - [Intraneural ganglion of the peroneal nerve]. AB - The intraneural ganglion of the peroneal nerve is a rare lesion. The differential diagnosis includes lesions of the knee especially the tendinopathia of the knee flexors. Clinical examination, electrophysiological investigations, ultrasound imaging and magnetic resonance tomography will establish the diagnosis. We report a case in which an artificial puncture of the ganglion caused a temporary peroneal palsy. Surgical procedure led to complete restoration of the nerve function. PMID- 9333975 TI - [Sudden loss of consciousness: clinical presentation and pathophysiologic mechanisms]. AB - Most cases of sudden and temporary loss of consciousness [syncope] are caused by hypoperfusion of the formatio reticularis. more rarely by primary neurologic or metabolic disorders. The most common etiology is vasodepressor (vasovagal) syncope, which is caused by peripheral vasodilation due to acute withdrawal of the efferent sympathetic tone, while the parasympathetic outpour is increased at the same time. Although the efferent limb of the baroreflex manifests in a rather uniform way, the afferent parasympathetic limb is very variable, leading to a variety of clinical presentations and triggers (orthostatic hypotension, pain, fear, cough, micturition, emotions). While vasodepressor syncope mainly occurs in young people with healthy hearts, cardiac syncope caused by arrhythmias or obstructive lesions are more frequently found in elderly patients with organic heart disease. Neurogenic syncope comprises either primary neurologic disorders, such as epilepsia, or hypoperfusion of the vertebrobasilar system (TIA). Rarely, an acute increase of intracerebral pressure may cause syncope. Similarly, metabolic disorders or side effects of drugs are rare causes of syncope; however, drugs may act as important cofactors in the pathogenesis of syncope. PMID- 9333976 TI - [Vasovagal syncope]. AB - Vasovagal syncope, also called neurocardiogenic syncope, is common with younger people. It results from an inappropriate, excessive autonomic reflex activity. In the elderly patient the syncope may be provoked by massage of the carotid bodies and is then known as carotid sinus syndrome. The pathogenesis of neurocardiogenic syncope is debated. Sudden vasodilation and/or bradycardia have been attributed to the activation of ventricular mechanoreceptors. The use of betablockers is based on this hypothesis. Head-up tilting at 60 degrees is helpful in the evaluation of syncope. In the therapy of recurrent vasovagal syncope, a thorough information of the patient and an adaptation of behaviour are often successful. Some authors have reported goods results with betablockade, etilefrin or mineralocorticoids. The patient with repeated severe syncopal attacks and asystole may benefit from an implantable DDD pacemaker. PMID- 9333977 TI - [Hemodynamically-induced syncope]. AB - Hemodynamic syncope is caused by an impediment to a necessary increase of the cardiac output; therefore, hemodynamic syncopes most often occur during or shortly after exercise. However, a syncope at rest does not exclude a hemodynamic cause. Moreover, arrhythmias which may directly lead to syncope or accentuate the hemodynamic impediment are often present in cardiac diseases causing hemodynamic syncope. Hemodynamic syncopes are responsible for 2 to 3% of all syncopes leading to medical evaluation. Of these, more than half are caused by aortic stenosis and about one quarter by pulmonary embolism. Other reasons are rare. Hypertrophic cardiomyopathy is more often associated with arrhythmic than with hemodynamic syncope. Syncope in primary pulmonary hypertension is often preceded by dizziness, epigastric distress and faintness. Since the medical therapy may lead to hemodynamic deterioration, it must be started under invasive observation. Primary tumors of the heart are rare; secondary cardiac neoplasms are 6 to 40 times more common. Myxoma is the most common primary tumor of the heart. It is important to promptly undertake surgery in order to improve prognosis. Various other diseases may provoke hemodynamic syncope; however, other symptoms are by far more common. PMID- 9333978 TI - [Syncopes from the neurologic viewpoint]. AB - A syncope is defined as a sudden, temporary loss of consciousness, associated with loss of postural tone with spontaneous recovery. The incidence is high and the differential diagnosis broad; therefore, the first observations are essential for the management of the patient. In this review diseases will be described which manifest themselves with syncopes that fall under the auspices of either internal medicine or neurology. First, the etiologies of syncopes are discussed in the strict sense of the word, i.e. due to a global cerebral ischemia such as in orthostatic hypotension and vasopressor syncopes. Thereafter, a discussion concerning the differential diagnosis of syncopes will be introduced, including mainly psychogenic and epileptic seizures as well as vertebrobasilar hypoperfusion. Depending on the reason of the loss of consciousness, it can be a common benign disorder or a severe life-threatening disease. The personal history or witness account of the incidence provides the most useful information concerning diagnosis. The cardiological diagnostic procedures are discussed elsewhere. In some instances an EEG can further help with the diagnosis. In many cases an etiology can't be found, even if extensive investigations have been performed. PMID- 9333979 TI - [Bradycardia-induced syncope]. AB - Bradyarrhythmias are the cause of syncope in 3 to 10% of cases. Marked bradycardia or asystole can be due to impaired function of the sinus node (sick sinus syndrome), high degree AV block or neurocardiogenic disorders (carotid sinus syndrome, vasovagal syncope). A precise history, ECG and 24-h. Holter recordings are the most helpful tools in diagnosis of bradyarrhythmia-induced syncope. An association between symptoms and documented ECG is essential for proper diagnosis. Sometimes an event-recorder may be helpful for this purpose. If a patient has normal physical examination, ECG, Holter, stress test, tilt-table test and echocardiography, no further electrophysiological investigation is needed. If the noninvasive tests show pathologic results that do not clearly explain the cause of syncope (sinus bradycardia, first-degree AV block, fascicular block, structural heart disease), then electrophysiological studies are recommended, which will also rule out ventricular tachyarrythmias in the differential diagnosis. If all diagnostic tests in a patient with syncope are normal, the prognosis is fairly good despite 30% recurrence rate. Treatment for symptomatic bradyarrhythmias is implantation of a pacemaker. The selection of an appropriate pacemaker system is very important. Dual-chamber systems (DDD) provide physiological stimulation by maintaining AV synchrony; thus, they should be preferred whenever possible. PMID- 9333980 TI - [Tachycardia-induced syncopes]. AB - In 50% of the patients presenting with a syncope, the cause is cardiac. The incidence of sudden death with 24% is high in this group. Since most of the tachycardia-induced syncopes are due to ventricular tachycardia (VT), a careful diagnostic approach must be used. The possibility of a VT to end in a fibrillation is great, especially in the presence of a organic heart disease, which leads to a bad prognosis in such patients. The aim of a careful anamnesis and clinical history is to establish the presence of a cardiac disease. A Wolff Parkinson-White syndrome, a long QT, an old myocardial infarction or a coronary artery disease (CAD) can be assessed by echocardiography (ECG). Stress testing is useful in evaluating a CAD and can possibly lead to a diagnosis when a VT or a supraventricular tachycardia (SVT) is induced. ECG is used to assess the cardiac ejection fraction and in the evaluation of a suspected right or left cardiomyopathy. The ambulatory ECG allow a diagnosis only in 2 to 3% of the cases. Nevertheless, the presence of more than 10 PVC/h and/or asymptomatic nonsustained VT is a predictor for sudden death in syncopy patients. Detection of late potentials has a sensitivity of 50 to 83% and a sensibility of 89 to 91% for the prediction of inducible sustained VT during electrophysiological studies (EPS) in patients with syncope. However, the usefulness of this technique is not fully established, since there is no significant difference in survival or recurrence of syncope between patients with and without late potentials. The EPS is an invasive technique and therefore used at the end of the investigations. The cardiovascular mortality is low (4%) in patients with a negative EPS. A treatment is mandatory in tachycardia-induced syncopes even when the cause is a SVT. Antiarrhythmic drugs are useful for the treatment of SVT. However, radiofrequency ablation of the accessory pathway is preferable, since the success rate is over 90%, and the side effects of chronic ingestion of antiarrhythmic drugs can therefore be avoided. Some VT can be treated successfully with drugs under the control of an EPS, but most of the patients must have the implantation of an internal cardiac defibrillator (ICD). PMID- 9333981 TI - [Sudden loss of consciousness in childhood]. AB - Sudden loss of consciousness in childhood presents itself usually as a syncope and occurs in approximately 15% of all children. Although syncope in the pediatric age group appears to be an isolated phenomenon with good prognosis, it could also be a manifestation of a life-threatening disease. Thus, even the first syncope should be evaluated in children. A thorough history and a careful physical examination with special attention to cardiovascular and neurologic abnormalities usually point to the etiology of the syncope and must result in the appropriate laboratory tests. About 50% of pediatric syncopes are non cardiovascular, 20 to 30% are cardiovascular, and 20 to 30% are of unknown etiology. The most frequent syncope in childhood is neurocardiogenic. It is important to distinguish neurocardiogenic syncope from cardiac syncope and epileptic seizures. Neurocardiogenic and cardiac syncopes can be discriminated from epileptic seizures by their usually shorter duration of unconsciousness and rare postsyncopal disorientation. Cardiac syncope is rare in children with a structurally normal heart and may be associated with Wolff-Parkinson-White syndrome, long QT syndrome or congenital AV block. Children with congenital heart defects and cardiomyopathies who present with syncope must raise a high degree of suspicion for a cardiac syncope. Cardiac syncopes often yield a poor prognosis with substantial percentages of sudden death; therefore, a vigorous attempt has to be made to diagnose and adequately treat cardiac syncope in children. PMID- 9333982 TI - [Psychiatry in comprehensive transition]. PMID- 9333983 TI - [Psychiatry of the family physician--historical highlights]. AB - For centuries the general practitioner assumed, among other functions, that of the alienist insofar as mental disorders were regarded as accessible to treatment at all. After 1800 "psychiatry" developed as a medical specialty essentially based on the growing number of psychiatric hospitals. Thus, the general practitioner was left with the treatment of less severe mental disturbances like neuroses and with the selection of patients to be treated by the specialists. The introduction of psychotropic drugs in the 1950s opened new possibilities in the field of out-patient psychiatry. PMID- 9333985 TI - [Psychiatric diagnosis and examination in family practice]. AB - Psychiatric diagnoses are made according to ICD-10 in order to standardize the diagnostic process. In most cases the psychiatric diagnosis will be based on a thorough history and the psychiatric examination which stresses the importance of a systematic approach to the later. The use of psychiatric scales and physical investigations can be occasionally usefull. PMID- 9333984 TI - [Differential diagnosis and therapy of delirium]. AB - Delirium (i.e., acute confusional state) is a frequent syndrome due to different exogenous (physical, chemical or biological) factors. The prevalence of delirium increases with higher age (e.g., signs indicative of dementia) and particularly in those patients with severe somatic illness. Symptoms of delirium are considerably uniform irrespective of different etiological causes. However, delirium is frequently underrecognised which may lead to several complications. The article summarizes data on symptomatology, aetiology and pathogenesis of delirium. In addition syndromes sharing phenomenology similar with delirium are discussed and treatment recommended. PMID- 9333986 TI - [Diagnosis and therapy of anxiety disorders]. AB - Anxiety disorders may be encountered by the medical practitioner in the form of phobias, panic disorder or generalized anxiety disorder. A phobia is characterized by a strong, irrational fear of a given object or situation, often resulting in avoidance behavior. Phobic patients usually respond well to cognitive behavioral therapy. Panic disorder, which is distinguished by recurring, unexpected attacks of fear not bound to particular situations, may also be treated with cognitive behavioral therapy and/or with clomipramin, benzodiazepines or selective serotonin reuptake inhibitors. Patients with generalized anxiety disorder, the main symptom of which is a persistent, free floating fear over a period of at least several months, may be helped through relaxation techniques, counseling and/or medication with low doses of sedating tricyclic compounds or short-term treatment with benzodiazepines. This article will describe anamnestic findings and the results of clinical examinations of patients with anxiety disorders. Factors to be considered in differential diagnosis will be discussed. PMID- 9333988 TI - [Diagnosis and treatment of alcoholism]. AB - Alcoholism is the single most important psychiatric and psychosocial disease in Switzerland. The average Swiss citizen consumes about 10 l alcohol/year, the equivalent of half a bottle of hard liquor/week. The expenses for alcoholic beverages exceed the expenses for mandatory school education. Alcoholism is a chronic disorder which in its course and approach to treatment can be compared with diabetes, hypertension, and asthma. Similar to other chronic diseases, therapy consists in counselling and medication. The relevant goal is improvement rather than cure. PMID- 9333987 TI - [Diagnosis and treatment of affective disorders]. AB - The rates of major depression (5-12%) are considerably higher than for bipolar disorder (ca. 1%). Depressive disorder is most frequent in general practice. Although general practitioners recognise and manage efficiently a large number of depressed patients, at any consultation about half the patients are not diagnosed. Recognising depression is made difficult by the frequency in general practice of presentations with somatic symptoms (masked depression) and of depression related to physical disorder. The best method for the general practitioner to overcome these problems is by using a relatively direct interview for the main specific symptoms of depression. The general practitioner has a key role in the management of depression and as a gatekeeper with a prime responsibility to make appropriate referrals to specialists. Counselling members of the family or friends and recommending self-help groups are important to improve the therapeutic compliance of the patients. PMID- 9333989 TI - [Drug dependence--diagnostic and therapeutic aspects]. AB - In Switzerland, dependence on these drugs reflects primarily the relationship between patient and doctor, due to the strict regulations. Drugs involved are in most cases benzodiazepines and other sedatives, less frequently stimulants including weight-control products and drugs containing opioids. Patients' characteristics and motivations include 1.) attempt for specific psychotropic effects, 2.) habituation developed during long-term treatment, 3.) overconsumption of drugs without seeking specific psychotropic effects. Clinical syndromes and therapeutic and preventive strategies are described. PMID- 9333990 TI - [Treatment of drug dependent patients in general practice]. AB - After a brief characterization of the actual situation of drug abuse in Switzerland, the general principles of methadone treatment and drug-assisted ambulant withdrawal in the family practice are introduced. They are suited for patients who mainly consume heroin and have a fairly stable social background. Both forms of treatment can very well be offered and carried through by general practitioners, if there is a clear agreement between the patient and the doctor about the therapeutic setting and guidelines. In every case close psychosocial support over a longer time span must be part of the treatment. For this longterm treatment, the continuous relationship with the general practitioner is ideal. PMID- 9333991 TI - [Psychoses (schizophrenias)]. AB - We are speaking with E. Bleuler about the "group of schizophrenic disorders". We describe the different symptoms, varieties of life-long courses and possible predictors of outcome. Currently, patients with schizophrenic disorders are frequently treated in Switzerland as outpatients by primary care physicians. The vulnerability models allows the recognition of different pathogenetic factors and also the integration of various treatment modalities, like pharmacotherapy treatment, family treatment and psychotherapy. PMID- 9333992 TI - [Somatoform disorders and their therapy]. AB - Somatoform disorders are frequent manifestations of psychosocial stress and present themselves in form of unexplained somatic symptoms. Diagnostics and treatment of somatoform disorders is mainly performed in order to uncover organic pathology. However, organic pathology could not be found, which leads to repeated investigations finally contributing to increase of health costs. Authors highlight pathogenesis of somatoform disorders, summarise the syndromes of this group of disorders and give recommendations as to management and treatment. PMID- 9333993 TI - [Eating disorders]. AB - Practitioners and psychotherapists see themselves more and more confronted to patients, specially women, with eating disorders. According to recent studies it can be estimated that the incidence of bulimia lies between 2% and 5%, of anorexia between 0.2% an 1%. The percentage of separate elements of the disorder is considered to be much higher, e.g. for binge eating at least 25%. At present around 3 to 6 years go by until patients contact a therapeutic setting for the first time. But the shorter the time between first appearance of the disorder and beginning of its treatment is, the better the prognosis. In order to improve treatment an interdisciplinary approach of practitioners, psychotherapists, dieticians and if needed other specialists should be strived for. Although obesity must be seen in a psychopathologic context in many cases, here we concentrate on the discussion of anorexia and bulimia. PMID- 9333994 TI - [The suicidal patient]. AB - Suicide prevention in the general practitioner's practice should not only be a topic for patients that are close to putting an end to their lives. It should start with the recognition of an emotional crisis or depressive symptoms. Such patients have to be asked about suicidal thoughts or plans. Depressive states have to be assessed very carefully and treated adequately. The Gotland study has clearly shown that competence in the recognition and treatment of depression can significantly reduce the frequency of suicide. PMID- 9333995 TI - [Diagnosis and therapy of personality disorders]. AB - Approximately 10% of the unselected population are affected with personality disorders, among the patients of psychiatrists and family doctors the quota goes up to 40%. Personality disorders comprise deeply ingrained and enduring behaviour patterns, manifesting themselves as inflexible responses to a broad range of personal and social situations. They are stable and lead frequently to subjective distress and/or to impaired social functioning. The division in subgroups is made on the reason of typical patterns of experience and behaviour, but overlapping between different subtypes is frequent. People with personality disorders often come into conflicts with their environment because of their maladaptive behaviour which lead to crises and need of intervention. Psychopharmaca can be given in such situations, but substances with an addictive potential like benzodiazepines should not be prescribed for a longer period. The long-term psychotherapy of personality disordered persons requires an individual planing after a careful analysis of the behaviour pattern and should focus on concretely defined and reachable aims. Personality disordered persons belong to the most difficult patients, their long-term treatment demands appropriate therapeutic skills. In the primary care family doctors therapy and support is important but several basic rules should be followed. PMID- 9333996 TI - [The family physician and his "patient with family". Significance of the family system in treatment of patients in general practice]. AB - From a systemic perspective the general practitioner and the patient form a "medical-therapeutic system", which has the same features as other social systems. As the patient is also a part of his family system there are (more or less detectable) interactions between these two relevant systems as well. Compliance-from a systemic point of view-is not only an indication of the functioning of the "medical-therapeutic system" but also a result of those interactions between two relevant systems. Clinical trials show that with his interventions the general practitioner helps in solving problems of difficult, blocked family relationships with the goal of an adequate coping strategy for all involved family members. PMID- 9333998 TI - [Origin and development of autopsy in Wurttemberg reflected by contemporary sources]. AB - The present study deals with the introduction and the development of coroner's inquest in Wurttemberg. It could be shown that the contemporary discussion about the incidence of apparent death became an important reason for the establishment of necropsy in 1824. In spite of legal backing the institution of necropsy remained disputed until the end of the Kingdom of Wurttemberg in 1918. Above all, the differing qualification of the coroners gave rise to criticism. Although the legislator provided a general qualification in surgery, most the coroners did not boast a special expert knowledge. During the time in view, the participation of barber surgeons in necroscopy was decreasing; the percentage of physicians also remained very low. In 1911, only 119 out of 1814 coroners were physicians-on the other side 641 representatives of post-mortem examination worked as small-holders or day-labourers, and another 33 persons even earned their daily living by grave digging. With regard to this, it is evident that the coroners in Wurttemberg formed a most heterogeneous group-both in social and in professional respect. PMID- 9333997 TI - ["Different from the others". Kraepelin's assessment of Hirschfeld's educational film. A contribution to the history of psychiatry of the Weimar Republic]. AB - In 1921 the Medical Court of Honour of the County Brandenburg and Berlin procured an expert opinion by the well-known psychiatrist Emil Kraepelin on the first sex education film "Anders als die Andern". The Berlin sexologist Magnus Hirschfeld had launched the film in 1919 to bring his scientific targets of abolition of section 175 StGB and the liberalization of sexual morality to a greater public. Kraepelin succeeded in banning the film because of its subject being a threat to population policy in post World War I Germany. PMID- 9333999 TI - ["Lingue di seripi", "serpents' tongues" and "glossopetrae". Highlights from the history of popular "cult" medicine in early modern times]. AB - In the 16th, 17th and 18th century "Glossopetrae", popularly known as "Lingue di Serpi", found on the Mediterranean island of Malta, were extensively used for medical purposes as antidotes. These fossil teeth, including specimens of the "Carcharodon Megalodon" (an extinct variant of the great white shark), were ground to powder or used as amulet pendants and "credence" and exported to pharmacies and shops in various cities of Europe. In antiquity, authors like Plinius or Solinus, excluding any religious connotations, had regarded "Glossopetrae" as objects "fallen from heaven on dark moonless nights". However, from the beginning of the 16th century the miraculous antidotic power of the specimens found at Malta was very strongly connected with the Pauline cult there. This cult owed ist origin to the excerpt of the shipwreck of the Apostle of the Gentiles on this island, as recorded in the New Testament. As in so many cases found in medieval and early modern medicine and pharmacy, the renown, collection, distribution and use of the antidote "Glossopetrae" or "Lingue di Serpi" was never limited to its real chemical and pharmaceutical properties. In the period of enlightenment and secular thinking mythic medicine as "Glossopetrae" had lost ist "magical" power. Consequently, with beginning of the late 18th century also the Maltese "Glossopetrae" featured in literature merely as exotic objects of curiosity or symbols of an age bound to medical superstition. PMID- 9334000 TI - [Traditional camel medicine in China]. PMID- 9334001 TI - [Veterinary medicine comment on camel medicine in Fan-mu tsuan yen-fang]. AB - This short paragraph tries to identify the camel diseases compiled in the old chinese text according to modern veterinary terms. Due to the specific terminology of the camel treatise and its overall scarce symptomatology the diseases are difficult to evaluate. The majority of them obviously deal with acute infectious diseases which manifest themselves under such symptoms as high fever, depression, anorexia, cachexia, diarrhoea, general weakness, etc. But there are some diseases and ailments which can be interpreted in modern terms my means of the symptoms, descriptions and cures, e.g. mange, paradontosis and wry neck syndrome. PMID- 9334002 TI - [Treatment and rehabilitation of post-traumatic tetraplegia in advanced age. A report of experiences]. AB - We report about the results of the treatment of patients older than 70 years with posttraumatic tetraplegia. In 7 out of 11 patients treatment in a specialised department lasting for many months makes it possible to live a subjective satisfactory life at home in spite of being severely handicapped. The goals and limits of rehabilitation of patients with posttraumatic tetraplegia cannot only be derived from the age but from the individual capability of the injured patient. PMID- 9334003 TI - [Self-induced injuries--surgical aspects]. AB - So far, psychiatric-psychoanalytic theories have been able to explain the phenomenon "self-injury" only unsatisfactorily. Moreover, the patients do not turn to a psychiatrist in the first place, but to surgeons, dermatologists, gynecologists or general practitioners. This is therefore an interdisciplinary problem. Since general medical knowledge is relatively unhelpful in diagnosing self-inflicted disease and its treatment, these patients often do not receive adequate psychiatric co-management or further care or indeed often get the chance to delegate the act of self-injury to the physician. In view of the sustained tendency for the disorder to chronify, this frequently results in severe, partly irreversible and sometimes iatrogenically co-induced physical impairments. In the final analysis, it also leads to enormous financial burdens for the agencies which bear the costs. PMID- 9334004 TI - [Joint surface replacement of the lateral tibial plateau using patella transplantation]. AB - A case of surgical reconstruction of the lateral tibial plateau after impression fracture (Type 41-B3.1) of an 47-year-old female is presented. Joint reconstruction was performed by using a free patellar graft. The 16-year follow up shows a very good functional outcome without subjective complaints. This case underlines that performing patellar autografting for surgical reconstruction of severe injury of the tibial plateau seems to be a promising alternative especially considering young patients in spite of avoiding total knee replacement. PMID- 9334005 TI - [Monitoring intracranial pressure in patients with severe craniocerebral trauma]. PMID- 9334006 TI - [Traumatically-induced compartment syndrome of the tibia. Ultrasound diagnosis for qualitative assessment of late sequelae for musculature after dermatofasciotomy]. AB - To determine the significance of sonography in evaluating long-term damage of muscle surgically treated for compartment syndrome 27 patients of the Department for Trauma Surgery, University Clinic Essen, Germany, were examined on their anterior lower limb after an average of 98 (43 to 154) months after trauma. They had had a fasciotomy for imminent (n = 15) or manifest (n = 12) compartment syndrome. Comparing the healthy side a qualitative grading (0 to 3) of the changes could be introduced reflecting the extent of the increase in echogenicity and the loss of the typical muscle texture. Gray scale histograms confirmed the qualitative grading. Patients with manifest compartment syndrome showed severe changes (grade 2 and 3). In imminent compartment syndrome 2 patients with grade 2 and 13 patients with grade 0 or 1 were found. The sonographical changes can be explained by the known pathomorphological changes after compartment syndrome (denervation, scarification). Sonography is useful in the evaluation of soft tissue after compartment syndrome. The results underline the demands of early fasciotomy in imminent compartment syndrome for prevention of damage of muscle and nerve. PMID- 9334007 TI - [Fractures after total knee joint endoprosthesis. Results of treatment with various stabilizing methods]. AB - In 20 patients with a fracture of femur or tibia 5.2 years after arthroplasty of knee the results of the operative treatment are presented. The results show that especially bone damaging diseases as rheumatoid arthritis, osteoporosis and the loosening of the endoprosthesis are favorable for the fracture during the follow up. The conclusion of the investigation shows that in younger patients the external fixation by plates and screws is the preferential treatment, in elderly people or comminuted fracture an internal fixation, also in combination with an additional osteosynthesis, allows a fast mobilization. The number of observed complications is higher than in primary knee arthroplasty, the full weight bearing 1st delayed. The rate of further operations and unsatisfactory results is also higher being affected by the high mean age of the operated patients (73.4 years). The possible use of a total femur implant must be discussed critically because only an individual production can avoid further damage of the parts of the joint that were not concerned by the fracture. PMID- 9334009 TI - [Long-term outcome of endoscopic CO2 laser surgery of cicatricial laryngostenosis in children]. AB - Laryngeal scars were removed in 183 children 3 months to 15 years of age. A total of 538 endoscopic CO2-laser operations were made. Stents were inserted in 22 cases. Good and satisfactory follow up (1-8 years) results were obtained in 78 patients, unsatisfactory outcomes were recorded in 34 patients. Long-term results demonstrate that CO2-laser surgery of laryngostenosis is safe for children of different ages in that it does not hinder adequate development of the larynx. PMID- 9334008 TI - [Antibodies against alpha-2 and gamma-interferons affect the course of juvenile respiratory papillomatosis in children treated with alpha-2 interferon preparations]. AB - 1-year or longer treatment of juvenile respiratory papillomatosis (JRP) with alpha-2 interferons (A21) leads to emergence of antibodies to A21 in 54.3% of the patients. The greatest percentage of the antibody positives (94.4%) was found among JRP patients treated with intramuscular recombinant human A21 (reaferon). The proportion of the antibodies carriers fell to 15.4% if reaferon was used as a component of the new preparation viferon. JRP patients have also autoantibodies to A21 and gamma-interferon (8.74 and 33.3%, respectively). Patients with autoantibodies to gamma-interferon responded well to treatment with A21. Antibodies to A21 seem to have no negative effect on interferon therapy of JRP. Reaferon and viferon can be effectively used in JRP treatment. PMID- 9334010 TI - [Treatment of neurosensory hypoacusis of antibiotic and non-antibiotic etiology with mydocalm and nootropil: comparison of efficacy]. AB - Mydocalm and nootropil combination was used for treatment of antibiotic neurosensory hypoacusia (24 patients) and non-antibiotic hypoacusis (11 patients). 100 mg of mydocalm was diluted in 10.0 ml of 0.9% sodium chloride and injected intravenously. 20% solution of nootropil (5.0 ml) was injected intramuscularly. The 14-day course produced subjective response. Noise in the ears and heavy head feeling disappeared. As shown by measurements of auditory thresholds, improvement of hearing was only subjective. The responses were similar in both groups of patients. PMID- 9334011 TI - [Ultrastructural study of the organ of Corti in animals given kanamycin and ceruloplasmin]. AB - 10 guinea pigs of group 1 were injected subcutaneously kanamycin (400 mg/kg). The other 10 guinea pigs of group 2 received human ceruloplasmin as an intraperitoneal injection of 10% solution (0.25 ml of the protein preparation per 100 g body weight) 30 min before kanamycin administration. 7 days after the treatment the cochlease were isolated and examined electron microscopically. The Corti's organ was stained with alcyan blue and lanthanum hydroxide. In animals of group 1 there was mitochondrial vacuolization, enlargement of the cisterns of the endoplasmic network, Golgi apparatus, sub-superficial network; glycocalix layer on the surface of the receptor epithelium was unbroken but uneven, in the sensor cells of the Corti's organ of the group 2 animals only few mitochondria were vacuolized. A great number of ribosomes and mitochondrial contacts with membraneous cell structures were indicative of active protein synthesis and energetic processes. Glycocalix was less damaged. This was indicative of an otoprotective effect of ceruloplasmin which diminished kanamycin ototoxicity. PMID- 9334012 TI - [Origin and physiology of the vestibular nystagmus]. AB - Oculomotor reactions in tracing moving visual objects by moving eyes and head were studied in 30 healthy subjects. With changing speed of the moving objects, electrooculogram registrating eye movements acquired the form of the vestibular nystagmus. The amplitude and frequency of the nystagmic movements were defined through the object's acceleration. The control tests for static fixation in active turns of the head and dynamic eye fixation nystagmic movements of the eye were absent. The findings support the hypothesis that nystagmus is a visually determinated reaction of the glance control system and that vestibular afferentation has no independent connection with oculomotor muscles. PMID- 9334013 TI - [Registration of the vocal field for the clinical assessment of voice]. AB - Phonatory function was assessed in 92 patients with various laryngeal diseases; polyps of the vocal cords, Reinke's edema, laryngeal papillomatosis (56, 17 and 19 patients, respectively). The control group consisted of 58 healthy subjects with normal voices. Principal acoustic voice parameters were measured using phonetograms: pitch range, maximum intensity range, area of the vocal field. The tests were made before and 10 days after endolaryngeal microsurgery. Marked postsurgical subjective voice improvement was achieved in most cases and was confirmed by acoustic measurements. Mean values of the above acoustic parameters demonstrated significant (p < 0.05) improvement after phonosurgery. The best functional results were recorded in the group with vocal cord polyps. The conclusion is made that quantitative assessment of phonetograms provides reliable information for clinical voice evaluation. PMID- 9334014 TI - [Computer multimedia technology in otorhinolaryngology]. AB - The author draws attention of ENT specialists to advantages of the novel computer system multimedia which is a promising tool for otorhinolaryngologists as it allows composition of the image, sound, text, drawing, animation, etc. The author describes the program and devices providing visualization of the ENT organs on the computer display. PMID- 9334015 TI - [Surgical treatment of congenital atresia of the external acoustic meatus in a child and audiological prognosis of its outcome]. AB - After detailed audiological examination a 11-year-old boy was surgically treated for a congenital complete left atresia of the acoustic meatus. The operation consisted of creation of the acoustic meatus, plastic reconstruction of the external opening of the acoustic meatus with one-stage tympenoplasty. The treatment provided good cosmetic and functional results confirmed by consequitive audiometric examinations. The operation was characterized by the absence of the antrum. PMID- 9334016 TI - [Computer tomography in the diagnosis of ear neoplasms]. AB - 51 suspects for tumors of the ear were examined using computed tomography (CT). Indications and parameters are specified for CT of the temporal bone as well as direct and indirect signs of benign and malignant tumors originating primarily from the middle ear. There are differences in the CT picture of ear inflammation with proliferation, destruction of tissue and tumors of the ear. CT findings were verified visually during surgical interventions. High diagnostic efficacy of CT was confirmed. PMID- 9334017 TI - [Efficacy of polymer suture material "kapromed" in otorhinolaryngological practice]. AB - When used in suturing, polymer material capromed containing antimicrobial substances proved its efficacy in laryngectomy, plastic reconstruction of pharyngo- and tracheostoma, management of operative wounds on the neck, especially in patients with laryngeal cancer preexposed to radiation treatment and other surgical interventions on ENT. Compared to conventional suture materials (silk, lavsan, catgut, capron), capromed improved the outcomes of surgical treatment, reduced the number of pyoinflammatory complications. PMID- 9334018 TI - [Extended trepanopuncture of the frontal sinuses (therapeutic and diagnostic opportunities)]. AB - The authors describe the technique of extended trepanopuncture of the frontal sinuses and relevant equipment (updated trephine, 4.2 mm wide-lumen drainage cannula, double-lumen attachment for it). The procedure enabled performing endoscopical review of the frontal sinus cavity, biopsies and minor operations through the lumen of the drainage cannula. The double-lumen attachment served for lavage of the frontal sinuses in the frontonasal obstruction. PMID- 9334019 TI - [Lipid peroxidation in the blood of children with pyoinflammation of the nose and paranasal sinuses]. AB - Lipid peroxidation (LPO) was measured in 84 children with pyoinflammatory diseases of the nose and paranasal sinuses (the study group) against 50 healthy subjects (the control group). LPO in the two groups was found different (the difference was statistically significant). Defects in LPO entail homeostatic disorders and indirectly cause impairment of oxidation-reduction processes. The efficacy of the conventional therapeutic measures can be improved by addition to the standard combined treatment of metabolic correctors which noticeably reduce possibility of the acute process transformation into chronic. Recurrences become less probable. PMID- 9334021 TI - [Tracheotomy in carbon monoxide poisoning]. PMID- 9334020 TI - [Tragus perichondrium and cartilage, temporal fascia in tympanoplasty]. AB - Plastic reconstruction of the tympanic membrane was performed with the use of tragus perichondrium and cartilage, temporal fascia in 245 and 141 patients, respectively, Tympanoplastic use of the tragus perichondrium and cartilage provides good morphological and functional results. Such surgery may be a method of choice in filling defects of the tympanic membrane. PMID- 9334022 TI - [Intubation granuloma of the cavum infraglotticum combined with limited chondroperichondritis of the cricoid cartilage]. PMID- 9334024 TI - [An otogenic cerebellar abscess in a child with favorable outcome]. PMID- 9334023 TI - [Asymptomatic long-term course of chronic otitis with marked destructive alterations]. PMID- 9334025 TI - [A large foreign body filling the nose, nasopharynx, maxillary sinus and pterygopalatine fossa]. PMID- 9334026 TI - [Ectopic salivary gland located on the palatine tonsil and simulating papilloma]. PMID- 9334027 TI - [Presentation of the patients in the course of clinical lectures]. PMID- 9334028 TI - [New method and instrument for maxillary surgery as an alternative to Caldwell Luc operation]. AB - A trocar of a novel design for maxillary surgery reduces to minimum operative injury of the cheek soft tissue and bone wall of the sinus providing an effective approach to sinus lesions. It enables widening of the sinus anastomosis by means of the diamond drill and forceps under microscope and endoscope control. The design of the instrument allows the surgeon to operate without the assistant's help. The duration of the operation is reduced, postoperative period runs uneventfully, with minimal negative responses of the cheek soft tissue and sublabial wound. Postoperative complications (subcutaneous emphysema and blood accumulation in the sinus) are corrected by irrigation or disappear spontaneously. The experience gained in operations on 180 patients and 248 sinuses provided the efficacy of the trocar in the removal of the polyps, foreign bodies, fungal masses and cysts both in children and adults. PMID- 9334029 TI - [Improving the command skills of student physicians during the review of a training assembly among the troops]. PMID- 9334031 TI - [The use of the water-pressure method in treating gunshot wounds]. PMID- 9334030 TI - [The results of delivering surgical care to the wounded and sick in military medical establishments and impending tasks]. AB - In article results of activity of the military surgeons on rendering of the surgical care to wounded and sick in 1996 are analyzed. During combat actions in Chechnya despite of severe forms of wounds and significant increase of combined battle traumas lethality among heavy wounded was reduced in 2 times. At common lethality rate in 1.3%, in hospitals from wounds 1.5% of wounded died, from traumas--0.7%, burns--2.9%, frostbitten--0.5%. As to peace time surgery, the analysis of main parameters of surgical work in military medical establishments, structure of diseases of servicemen, surgical activity, average terms of treatment, lethality after operations, defects in rendering of the surgical care is given. In conclusions the authors say about problems, that the military surgeons have today. PMID- 9334032 TI - [Dynamic spinal traction in the combined treatment of the pain syndrome in posttraumatic osteochondrosis]. PMID- 9334033 TI - [Means to improve therapeutic care in the Armed Forces]. AB - The article describes dynamics of diseases of therapeutic field in the Armed Forces in 1991-1996. The analysis of therapeutic sick rate has confirmed, that a major problem of military therapy remains acute pneumonia, which sick rate in 1993-1996 has increased twice. The sick rate of ulcer has increased equally. The main directions of perfection of organization of the therapeutic care in the Armed Forces are etiologic diagnostics and prophylaxis of acute pneumonia in the soldiers, creation of system of prophylaxis, active revealing and centralized high-specialized treatment of the patient with ischemic heart disease among officers, perfection of system of rendering of the specialized care to the patient of endocrinological field, introduction of modern methods of anti-relapse treatment of ulcer, active revealing of the persons, infected by virus of hepatitis, perfection of work of an out-patient department link. PMID- 9334034 TI - [The programmed therapy of rhythm and conduction disorders in ischemic heart disease]. PMID- 9334035 TI - [The importance of monitoring arterial pressure in the diagnosis and treatment of hypertension]. PMID- 9334036 TI - [The first and foremost tasks of the medical service]. AB - Now in connection with common situation in Russian Federation the problem of reinforcements of army and fleet by healthy personnel, scare of a call-up quota and its poor quality are the main problems of the Armed Forces at the state level. The uniform complex program of medico-social maintenance of the citizens during preparation for military service is necessary. The modern situation is difficult due to many infectious diseases, so the role and the place of military medical service grows. In last years structure of quota, served by the military doctors, and number of other parameters have greatly changed, that require revision of some priorities. A problem of reinforcements of the Armed Forces by medical service officers remains actual, for decision of which a full-bodied admission on military medical faculty is required, as well as admission of the officers under contract and calling-up of reserve officers. In article the main lessons, received by the medical service during combat actions in Republic of Chechnya are also formulated. PMID- 9334037 TI - [The diagnosis of pemphigus vulgaris in an isolated lesion of the oral cavity]. PMID- 9334038 TI - [The organizational bases and functional directions in the activities of the hospital epidemiologist]. PMID- 9334039 TI - [Current problems in health protection for servicemen]. AB - The important directions in the field of preservation and strengthening of health of the servicemen are creation of an effective prophylaxis system in the troops on the basis of command and all services efforts integration, and interaction with other departments and the local authorities, and also perfection of medical maintenance organization of the Armed Forces and its preventive component. The analysis of health risk factors permits to distinguish the factors of water supply, municipal and household services, food. As priority such directions, as organizational perfection of management of water supply, municipal and household services of garrisons, practical work in interests of the particular person, perfection of system of preventive medical measures should be considered. PMID- 9334040 TI - [Experience in the realization of diagnostic and treatment algorithms in ischemic heart disease and arterial hypertension]. PMID- 9334041 TI - [At the sources of Voenno-meditsinskii zhurnal]. PMID- 9334043 TI - [The 40th anniversary of the Department of Military and Emergency Medicine of the Russian Medical Academy of Postgraduate Education]. PMID- 9334042 TI - [An outstanding Soviet neuropathologist (on the centenary of the birth of A. V. Triumfov)]. PMID- 9334044 TI - [The use of multichannel electromyostimulation in the combined sanatorium-health resort treatment of children with the sequelae of birth-related brachial plexitis]. PMID- 9334045 TI - [The use of stimulation by sinusoidal modulated currents in the rehabilitation of children who have had poliomyelitis]. PMID- 9334047 TI - [The effect of deep massage and physical exercises on the cerebral circulation in osteochondrosis of the cervicothoracic spine]. PMID- 9334046 TI - [The duration of the sanatorium-health resort treatment of patients with neurological manifestations of spinal osteochondrosis]. AB - Patients with neurological symptoms of spinal osteochondrosis in exacerbation took reduced intensive courses of physiotherapy which consisted of ultraphonophoresis of 1% oil solution of polar fraction of lipids from sulfide mud (eplir). The course included 12 daily procedures. Positive functional changes and those in vertebroneurological status support high efficacy and good tolerance of such short courses. PMID- 9334048 TI - [The optimization of physiotherapy for patients with peripheral neuropathies of traumatic origin]. PMID- 9334049 TI - [The use of sapropels in osteoarthritis in participants in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. PMID- 9334051 TI - [Microwave therapy in the combined treatment of chronic secondary pyelonephritis in children]. PMID- 9334050 TI - [A quantitative evaluation of the action of EHF therapy in patients with leg fractures]. PMID- 9334052 TI - [The use of the polymerase chain reaction (DNA diagnosis) in the diagnosis of urogenital infections at a health resort and its medical economic efficacy]. PMID- 9334053 TI - [The possibilities for regulating thymus cell activity with the transcerebral use of physical factors]. AB - The authors discuss experimental findings on physical factors (impulse currents of different frequencies, SHF and IR laser radiation, coherent and non-coherent red light) effect on thymic cell genome in transcerebral exposure in models of some diseases running with immune disorders. PMID- 9334054 TI - [An inventory of natural therapeutic resources]. PMID- 9334055 TI - [The climatic and recreational potential of the Far East seaside region]. PMID- 9334056 TI - [Current problems in pelotherapy]. PMID- 9334057 TI - [The effect of heavy metals on the health-index microflora in mineral waters]. PMID- 9334058 TI - [The combined therapy of osteochondrosis of the cervical spine]. PMID- 9334059 TI - [Reactive arthritis and its treatment with physical factors]. PMID- 9334061 TI - [The certification of specialists for the physiotherapy service]. PMID- 9334060 TI - [The current aspects of the pelotherapy of patients with rheumatoid arthritis]. PMID- 9334062 TI - [New developments in standards and statutes legislation in the area of health resort medicine]. PMID- 9334063 TI - [The effect of transcerebral electrical impulse exposures on regional hemodynamics after the surgical treatment of uterine myoma]. AB - After surgical treatment of uterine myoma 28 patients with cardiovascular diseases experienced disturbances of cerebral, upper limb and small pelvis circulation. The course of transcerebral electrotherapy (short bipolar nonsymmetrical impulses with large amplitude of the negative part) relieved the symptoms as a results of better cerebral and upper limb hemodynamics. PMID- 9334064 TI - [The hypertensive type II diabetic patient treated with captopril in free general practice (Austrian Safety Study). An indications study]. AB - In 826 hypertensive patients including 396 with non-insulin dependent diabetes mellitus safety and efficacy of captopril 50 mg per day was evaluated throughout three months. In all patients blood pressure was significantly reduced. Moreover, in part of the patients with microalbuminuria, these tests turned negative with treatment. In addition, in patients with diabetes fasting and postprandial plasma glucose levels as well as HBA1C levels decreased. Only in 6.8% side effects occurred. In all patients quality of life as evaluated by a 10 item rating scale questionnaire improved. Taken together the results of this observational study confirm improvement of blood pressure levels, kidney function and metabolic derangements in diabetic patients treated with the ACE-inhibitor captopril. Effectiveness of these actions of captopril in respect to longterm prognosis in diabetics, however, remains to be established. PMID- 9334065 TI - [Technical aspects of studying motor asymmetry]. AB - The present review shows the variability assessing motoric asymmetries. There are preference and performance tests. Motoric asymmetries are strongest and most manifest for handedness (hand preference and performance), descending through footedness and other less known asymmetries. A vast range of testing techniques have been used to assess handedness. Writing and self-report are two of the most popular techniques. Other preference measures include observation of how people use tools and questionnaires. Performance test assess speed and accuracy in tasks stressing manipulative dexterity. Although questionnaires are generally thought to be reliable and valid instruments, there is a disagreement as to the nature, the number and weighting of the items to be included. PMID- 9334066 TI - [Recording scientific literature--problems with citation]. AB - The present review describes some problems of citation of medical articles basing on own experiences. The citation and bibliography should be carefully basing on the several given instructions. PMID- 9334067 TI - [Adjusting to illness and secondary prevention in unemployed patients from the medical psychology viewpoint]. AB - Due to the demands on medical and psychosocial facilities, an above average accumulation of unemployed patients was investigated. General practitioners and internists have reported high stress experiences of patients and the difficulties encountered with their illness reflection. It was clear in the examinations, that there was no fidderence in the way unemployed and employed reflected their illness, although a difference was shown in their coping of critical life experiences. In scope of the medical conversation, unemployed patients could convey specific coping abilities as a secondary prevention. PMID- 9334068 TI - [Painful hip in children--a differentiated view of coxitis fugax]. PMID- 9334069 TI - [Arthrosis]. PMID- 9334070 TI - [Incidence and prevalence of cox- and gonarthrosis in the general population]. AB - AIM OF STUDY: Analysis of data on the incidence and prevalence of osteoarthritis (OA) of the hip and knee in the general population. METHOD: Review of relevant investigations concerning epidemiology of hip and knee OA. RESULTS: Estimates of incidence and prevalence of OA of the hip and knee vary considerably among 29 studies from 14 countries and 4 ethnic groups (incidence: 10 to 2230 per 10(5) person-years, prevalence: 0.5% to 36%. DISCUSSION: Because of the differences in study design, study populations, and the definition of OA, a comparison between different studies is very difficult. Nevertheless, the following patterns can be identified: In general, both the incidence and prevalence of OA of the hip and knee increase with age. The sharp increase in incidence of symptomatic OA of the knee after age 50 in women suggests an influence of menopausal changes of hormonal status. The prevalence of radiographically defined OA of the hip is higher in men than in women, but a reverse pattern was found for OA of the knee, especially after age 45. The prevalence of radiographically OA was found to be higher in caucasian than non-caucasian populations at the hip but not the knee joint. Among patients with radiographically defined OA at the hip, joint pain seems to be more common in women than in men. In contrast, the frequency of joint pain in persons with radiographically defined OA of the knee is about the same in women and men. PMID- 9334071 TI - [Standardization of roentgen diagnosis in coxarthrosis and gonarthrosis in clinical studies. Recommendations of the 1st Working Circle of the DGOT (Connective Tissue Research and Arthrosis Deformans)]. AB - AIM OF STUDY: Recommendations of standardized radiographic assessment in clinical and epidemiological studies on patients with hip and knee osteoarthritis. METHODS: Survey of published studies and results of consensus conferences regarding reproducibility of radiographic assessment and standardization procedures. CLINICAL RELEVANCE: With the proposed guidelines a standardized and reproducible radiographic assessment of patients with hip and ankle osteoarthritis in clinical and epidemiological studies is possible. The recommended procedures can be supplemented by additional techniques. PMID- 9334072 TI - [Reproducibility of radiologic diagnosis in gonarthrosis]. AB - AIM OF STUDY: Ongoing efforts of the "German Society of Orthopaedic Surgery and Traumatology" (DGOT) to standardize diagnosis and therapy of osteoarthritis, necessitated this study, where the reproducibility of different radiographic features of knee-OA was assessed. METHODS: Three readers graded 100 antero posterior and lateral knee radiographs for selected features (femorotibial osteophytes, joint space narrowing, sclerosis and chondrocalcinosis; patellofemoral osteophytes) and an overall-score (Kellgren and Lawrence, 1963) at two time points 3 months apart. Intra- and inter-observer-reliability were calculated by intra-class correlation-coefficient (ICC). RESULTS: Osteophytes in the femorotibial as well as in the patellofemoral joint could be assessed with a high intra- and inter-observer-reliability. While for joint space narrowing intra observer-reliability is excellent, the inter-observer-reliability is less satisfactory, and subchondral sclerosis as well as chondrocalcinosis showed even less reproducibility. The Kellgren-Lawrence global score proofed to be highly reproducible. CONCLUSION: Based on the results of this examination, we can recommend a reliable radiographic classification of knee osteoarthritis by grading of relevant individual features (osteophytes and joint space narrowing) and overall assessment. PMID- 9334074 TI - [Shape transformations of the lumbar spine in relation to passive extension of the lower extremities in the sagittal level]. AB - PROBLEM: The interdependencies between movements of the thighs and the lumbar vertebral shape are of high practical interest. Which are the normals of this phenomenon? METHOD: In an experiment on 107 volunteers without before known spinal disorders and complaints of back pain (47 f, 60 m, 17 a-30 a), the interdependencies between movements of the thighs in the sagittal and the lumbar back profile were analysed. Hip joint movements were provoked by a lift jack, elevating the feet to the volunteers, which sat on a bicycle chair. The hip joint flexion was measured by a Zebris CMS 50. The sagittal profile of the lower back was sensed by a comb of steel needles with low friction support. RESULTS: At 30 degrees of hip flexion, 68% of the volunteers demonstrated a kyphotic, 17% a straight and 15% a lordotic lumbar shape. Starting at 90 degrees of hip flexion, "definitively kyphosating movements" of the lumbar motion segments occur. At the end of the motion, 89% of the volunteers had a kyphotic, 3% a straight and 8% a lordotic lumbar configuration. Each 2 degrees of additional hip joint flexion caudo-cranially one more lumbar motion segment is recruited for the definitive kyphosation of the lumbar spine. CONCLUSIONS: Instead of a "physiological shape of the lumbar spine" its "physiological function" or its "physiological interaction between shape und function" should be in the focus of future discussions. In the sitting, hip joint flexion leads to a coupled motion of the thighs, the pelvic girdle and the lumbar vertebral column with the consequence of a kyphosation of the lumbar back shape. PMID- 9334073 TI - [Spatial orientation and postural regulation in patients with idiopathic scoliosis]. AB - QUESTION: Etiology of idiopathic scoliosis is unknown. In literature a primarily neurogenic disturbance of postural regulation is discussed with subsequent changes of the spine. In the present paper, individual functions of postural regulation in patients with idiopathic scoliosis were examined by neurophysiologic investigations and compared to a normal population. METHOD: The body sway of 28 patients was investigated in upright position under various testing conditions by means of a force measuring platform, allowing examination of the efferent part of postural regulation as well as the sensory systems involved. Registration of the eye movements (ENG) made it possible to investigate the function of the vestibulo-cerebellar system. Furthermore the optokinetic capability of scoliotic patients was tested by determination of the subjective visual vertical (SVV). RESULTS: It turned out that none of the neurophysiologic test procedures showed clearly pathological findings in scoliotic patients as compared to a normal population. CONCLUSION: It has not to be considered as probable that idiopathic scoliosis is caused by a disturbance of postural regulation. PMID- 9334075 TI - [Determination of bone quality before spinal instrumentation--value of different in vivo methods]. AB - OBJECTIVE: The present investigation should elucidate which assessment technique for bone quality is most appropriate to estimate preoperatively fixation strength of instrumental spine fusions. METHODS: VDS-screw fixation strength in 50 human cadaveric vertebral bodies was approximated by means of pullout force assessment. Bone quality was assessed by Dual Energy X-ray Absorptiometry (DEXA), Quantitative Computed Tomography (QCT), MRT and histomorphometry. For each of these techniques, correlation with axial pull out force strength was investigated. RESULTS: Highest correlation was found for cancellous bone density (QCT) (r = 0.72; p < 0.001) and DEXA (r = 0.70; p < 0.001). MRT, cortical bone density (QCT) and histomorphometry just slightly correlated with pullout force strength. CONCLUSIONS: Absorptiometrical techniques like QCT and DEXA are most appropriate to estimate VDS-screw fixation strength preoperatively. PMID- 9334076 TI - ["Primary" shoulder stiffness: illness duration and therapeutic comparison]. AB - AIMS: Etiology, natural history and therapy of the frozen shoulder still remains obscure. Therefore observation of natural history is of interest. METHOD: In a retrospective study 140 patients with different therapies were followed-up. RESULTS: 28 (20%) patients were not considered as healed because of persisting complaints during the whole follow-up period with an average duration of 49 months. Mobilisation under anaesthesia (27 patients) showed an less improval in range of motion with a shortening of complaint period. CONCLUSION: The existence of a subgroup of patients with no response on regular therapy is assumed. PMID- 9334077 TI - [Arthroscopic trans-glenoid stabilization of post-traumatic anterior shoulder instability]. AB - PURPOSE: Aim of this study was to evaluate the results after athroscopic transglenoidal stabilization in patients with anterior posttraumatic shoulder instability. METHODS: 30 patients with posttraumatic anterior shoulder instability were prospectively observed for a mean of 36 months (24-56) after an athroscopic stabilization has been performed. The operative technique was carried out as described by Morgan with use of transglenoidal sutures to repair the labrum. RESULTS: All patients had a Bankart lesion and a Hill-Sachs defect. According to the criteria of Rowe, 24 patients (80%) had good or excellent results and 1 patient (3%) was graded as fair. 5 patients (17%) developed recurrent instability 6-24 months postoperatively so they had failed results. 83.4% had no or little limitation in sports activity. Sex, age or grade of activity had no influence on the result concerning stability. The mean preoperative dislocation rate was 8.6 for the failures and 5.9 for the stable results (p < 0.05). CLINICAL RELEVANCE: The results of arthroscopic stabilization of the shoulder are inferior to the classical open repair. It should only be performed in patients with unidirectional, posttraumatic anterior shoulder instability without capsulaligamentous hyperlaxity or multiple resdislocations. PMID- 9334078 TI - [Prosthesis edge nodule and verrucous hyperplasia in leg amputees]. AB - Dermatologic problems in leg amputees mainly result from their specific prosthetic supply. As this relationship is not well documented in the German language literature, we try to illustrate important pathophysiologic aspects of lower and upper limb prostheses in two highly characteristic dermatoses, epidermoid cysts and verrucous hyperplasia (papillomatosis cutis lymphostatica). Their management includes improvements in prostheses' workmanship and conceptional changes (e.g. CAT-CAM-technique). Interdisciplinar diagnostic and therapeutic efforts particularly between orthopedic surgeons, prosthetists and dermatologists should prevent immobility of the amputee at an early stage. PMID- 9334079 TI - [Diagnosis and therapy of psoas abscess]. AB - PROBLEM: Psoas abscesses are really rare so that the diagnostic onset is commonly very late. The differential diagnosis to other retroperitoneal processes is therefore important. METHOD: In a period of observation of 6 years 21 patient were treated with psoas abscesses. The evaluation of 16 records was done retrospectively under consideration of etiology, history, clinical examination, lab results and x-ray/CT/MRI etc. RESULTS: With the knowledge of the anatomy of the ilio-psoas muscle the clinical examination gives us important information about the diagnosis of psoas abscess. The history and the clinical examination precede the further diagnostics and are condition for high rates of sensitivity and specificity. Lab results indicate an absedation without being specific. The exclusive position of radiological diagnostics is undisputed. Ultrasound, x-ray and leucocyte marked bonescan are proven to be helpful in cases of unknown location of the abscesses. Method of choice seems to be the contrast enhanced CT scan. The differential diagnosis includes gastrointestinal or renal disorders as well as pathology of bone or joints. In our cases differential diagnosis was complicated since the diagnostic onset was delayed and the initial therapy was not adequate. The diagnosis "abscess of the psoas" does not imply a general regime for therapy therefore an individual treatment in consideration of percutaneous and operative drainage has to be recommended. In selected cases a combination therapy is advised. CONCLUSION: In every case of retroperitoneal symptoms the differential diagnosis of an abscess of the psoas has to be regarded. The diagnosis is subtil and requires clinical and laboratory examinations as well as contrast enhanced computerscan. The therapy follows operative measures. The technique has to be individually decided. PMID- 9334080 TI - [Perthes disease--results of a containment-oriented therapy concept]. AB - INTRODUCTION: In a retrospective study a treatment concept for Perthes' disease dependent on the containment was applied. PATIENTS/METHODS: 49 hips of 41 children (9 female, 32 male) were treated between 01. 01. 1990 and 31. 12. 1995. In our concept of treatment a varus femoral osteotomy was performed in 28 cases with not contained hips or less than 4/5 coverage of the femoral head (X ray/MRI). The other 21 well contained hips with 4/5 coverage or more were treated conservatively with physiotherapy and in case of joint effusion and pain additionally with the use of crutches (partial weight bearing) and anti inflammatory medication. The average age in the non-operative group at the time of first investigation was 4 years and 9 months (3 y./1 m. to 7 y./1 m.) and 6 years and 3 months (4 y/2 m. to 10 y/0 m.) at our last examination (mean follow up 17.7 months, range of 6 to 72 months). At the time of indication for a varus femoral osteotomy the patients had an average age 6 years and 1 month (3 y./6 m. to 10 y./2 m.), the mean age at the last postoperative examination was 7 years and 11 months (4 y./8 m. to 12 y./5 m.) with an average follow up of 21.5 months (6 to 77 months). RESULTS: For the conservatively treated children we achieved good results (still well contained hips with 4/5 coverage, no decrease of function, no increase of pain) in 85.7% (18 of 21 cases). In 85.7% (24 of 28 cases) we found good results (well contained hips, increase of coverage, no decrease of function, no increase of pain) in the operation group. CONCLUSIONS: The presented concept of therapy in Perthes' disease was practicable for all patients and included the possibility of decision for operative or non-operative treatment. In both groups we achieved good results in 85.7% of the cases. PMID- 9334081 TI - [Autologous transfusion in total hip endoprosthesis--a clinical concept]. AB - The risks associated with blood transfusion can be minimized using autologous blood. The efficiency of preoperative blood deposit, preoperative acute hemodilution spinal anesthesia, controlled hypotension and intra and postoperative autotransfusion in reducing homologous transfusion has been demonstrated. In a prospective study the effectivity of this concept is demonstrated. 141 patients scheduled for total hip arthroplasty were divided in three groups: group I--total hip arthroplasty without cement (n = 55); group II- total hip arthroplasty with cement (n = 52); and group III total hip revision arthroplasty (n = 34). The mean quantity of the donated blood was 900 mL. The donation was not associated with serious complications. In group I 52 patients (94.5%), in group II 40 patients (77%) and in group III 14 patients (41.2%) did not require homologous transfusion. The difference between group I and II was significant (p < 0.05). Under the conditions studied, preoperative autologous blood deposit, acute hemodilution, spinal anesthesia and controlled hypotension are effective for decreasing the application of homologous transfusions in hip arthroplasty. The efficiency of preoperative hemodilution alone is limited. Preoperative deposit of autologous blood is a simple, effective, economical, and low-risk method of reducing homologous transfusion. Autotransfusion with a cell separator can save approximately 50% of the erythrocytes lost during revision total hip arthroplasty. PMID- 9334082 TI - [Paget's disease of the proximal ulna--a rare monostotic manifestation]. AB - Bone changes of the upper extremity are rare manifestations of the M. Paget disease. Most frequently the disease affects the lower extremities, the pelvis followed by the calvarium and others. The rare asymptomatic, monostotic manifestation of Paget's disease of the left proximal ulnae of a 59 year old woman and the way of diagnosis is described. Symptomatic skeletal lesions are detected by radiological examination, bone scintigraphy reveals asymptomatic lesions. Laboratory tests (Alkaline serum phosphatase, Hydroxyproline in urine) monitor the activity of the disease. PMID- 9334083 TI - [Ultrasound imaging of the patellar tendon--an experimental study]. AB - QUESTION: The accuracy of ultrasound in the detection of histological changes is a point of discussion. METHOD: In an experimental study we examined on 58 explanted human patellar tendons the correlation between histological and sonographic changes. RESULTS: Ultrasonographic measurements of length, width, thickness and circumference of the ligaments correlated highly to the explants' macroscopic evaluated dimensions. The sonographic lesions were distinguished in changes in form and echogenicity. Macroscopic visible changes in the explanted ligaments could be detected ultrasonographically. In histological aspect 2 tendons and in ultrasonographical aspect 16 tendons were found to be normal in the whole course of the tendons. 14 ligaments showed histological but not ultrasonographical abnormalities. 42 ligaments were conspicuous in both methods of examination. In the course of the tendons a very high correlation (up to 92%) between sonographical and histological findings was detected. At the insertion are of the patellar ligament the correlation was low. CONCLUSION: The performed study shows that ultrasonographically detected lesions of the patellar ligament point to histological changes but no conclusions to the kind of histological changes can be taken. However ultrasonographically found abnormalities correlate in a high proportion to histologically detectable changes in the ligaments. Ultrasonography is a good method to evaluate degenerative symptoms in the patellar ligament. PMID- 9334084 TI - [The SAPALDIA Study--no results?]. PMID- 9334086 TI - [Occupationally-induced degenerative discopathies in the area of the lumbar spine]. AB - A case-control study comprising 188 male and 160 female cases from an orthopedic's cabin explained occupational risk factors for osteochondrosis, spondylosis, and spondylarthrosis of the lumbar vertebra. The controls (109 men, 130 women) steaming mainly from other physician's offices were free from musculoskeletal symptoms. Age-adjusted odds ratio (OR) were calculated for both sexes. In women elevated ORs were found for working in standing position (OR 2.2 CI 1.31-3.85), for repetitive work (OR 1.9 CI 1.17-3.36), for lifting heavy things up to 20 kg (OR 2.1 CI 1.24-3.67), for working as a secretary (OR 1.8 CI 1.16-3.0) or in a supermarket (OR 2.1 CI 1.05-4.56). Men showed elevated ORs for bending (OR 2.2 CI 1.27-4.08), vibrations of the upper limbs (OR 2.3 CI 1.12 5.02), whole-body-vibrations (OR 2.1 CI 1.14-4.11), working in the metal industry (OR 3.6 CI 1.88-6.89) or as motorists for more than ten years (OR 6.5 CI 1.54 27.99) and working in a humid or cold environment (OR 2.2 CI 1.30-3.72). PMID- 9334085 TI - [Measles, mumps and rubella vaccination status of school beginners in Munich]. AB - A cross-sectional study was performed on all of 10029 school-beginners in Munich in 1994 to investigate the vaccination status of measles, mumps and rubella immunisation. The objective of the study was to determine socio-demographic and psychological factors affecting the MMR vaccination rate. Data were received from 81.8% of all 10029 school-beginners. The vaccination rate was 86.1% for measles, 84.5% for mumps and 72.9% for rubella (missing values not included). Low overall vaccination rate was found in not first-born children, children of parents with non-german nationality, in children of parents with a low socio-economic status, in children accompanied by a working parent, and in children accompanied by a smoking parent. Children without any denomination also showed a lower vaccination rate. Girls had a higher vaccination rate for rubella than boys. A higher overall vaccination rate for MMR was associated with parents considering these infections to have a high impact for people's health. Vice versa children of parents considering adverse effects of vaccination against MMR as an important impact on health had significant lower vaccination rate. The results of a multiple logistic regression model showed two factors significantly affecting the MMR vaccination rate: Physician's recommendation and individual attitude towards medicine seem to have the most important influence on decision making for or against vaccination. In conclusion MMR vaccination strategies have to be improved. New ways such as, no vaccination--no school's should be considered for Germany. PMID- 9334087 TI - [Mortality and birthdays]. AB - The authors study the date of death in relationship, in the annual cycle, to date of birth of every person who died of natural causes in Switzerland between 1st January, 1969 and 31st December, 1992 (N = 12,275,033). They highlight a maximal mortality (17% over expected) on the actual anniversary of birth with the entire population, a slightly lower figure being found under 80 years of age, higher over this age. They tie this phenomenon to the classical group of "anniversary reactions", in which problems of identity probably play a role. An uncertainty hangs nevertheless over the value of these findings, which are not confirmed by any published material, and even denied by some. PMID- 9334088 TI - [Determinants of the quality of life in chronic renal failure]. AB - Dialysis patients, waiting for kidney transplantation, were asked about their quality of life. Data from 1027 persons have been collected. Compared to a population sample by the "Nottingham Health Profile" (NHP), dialysis patients showed double the frequency of symptoms--only for the subscale "pain" no significant difference could be recognised. Duration of dialysis treatment reduces the quality of life considerably: increasing troubles have been observed through different quality of life scales. Age shows less important influence concerning "pain" and "physical mobility", even a decrease of symptoms in elder patients has been demonstrated by NHP-subscales for "emotional reaction" and "social isolation". Gender, education, kind of disease and dialysis treatment, and the fact of former transplantations had only marginal influence on some different dimensions of life quality. The study demonstrates in which way the patients perception of life quality could be operational and integrated in analysis and evaluation of therapeutic procedures. PMID- 9334089 TI - [German version of the Nottingham Health Profile (NHP): translation and psychometric validation]. AB - The Nottingham Health Profile was developed in the late 70s in Great Britain and has since then been widely used in anglo-american countries as well as in other countries. We report about the translation process and the examination of psychometric properties of the German version of the Nottingham Health Profile in 10 samples of over 1000 healthy and ill persons. Results of psychometric assessment (feasibility, reliability, validity, sensitivity) were satisfactory. This suggests that the German translation of the Nottingham Health Profile is a reliable and valid instrument for patient-based reports of subjective health which can be used in clinical research as well as in epidemiologic studies. PMID- 9334090 TI - [Response rate and analysis of non-responses in a cohort study]. AB - In 1987, a national survey on psychotropic substances and life styles was carried out by the Swiss Institute for Alcohol and Drug Problems (SIPAD). Out of 1910 participants, 1568 did agree to take part in a further study. In 1995, in order to execute a cohort type survey with minimal cost, the co-ordinates of 1472 of them were retrieved and a new questionnaire was sent to them. After 2 written reminders and another one by phone, the response rate reached 72.6%. It is therefore possible, at least in Switzerland, to lead a cohort type survey using a cross-sectional study as the starting point. Significant differences arise between the respondents and non-respondents at the level of linguistic region, age, income, civil status, educational level and alcohol or drugs consumption. On the other hand, neither sex, nor tobacco or cannabis consumption had an influence on the response rate. PMID- 9334092 TI - [Quality of life after pneumonectomy for bronchial carcinoma]. AB - Quality of life measurements gained increasing importance in the last years. After lung tissue reducing interventions for bronchogenic carcinoma quality of life measurements play an important role, since quality of life can be decisively influenced by post-operative reduced lung function. On the basis of postoperative physical symptoms and lung function the restriction of quality of life after pneumonectomy should be analyzed with respect to intervention and adjuvant therapy, 36 patients with an average age of 61 years were followed up by ambulatory oncological care for 40 months (median) after operation and lung function as well as quality of life were measured by self assessment index QLQ- C 30 of EORTC. RESULTS: Beside physical symptoms (increasing of dyspnea by 61.1% and pain by 30.6% after pneumonectomy as compared to preoperative values) the significant reduced lung function (IVC by 33.5%, FEV 1 by 27.1%) and the QL measurement showed the greatest restrictions, latter in "Physical functioning" by 27.2%, in "Role functioning" by 45.8% and in "Global health status/ Quality of life" by 45.6%. The symptoms "Fatigue" (Score 39.8), "Dyspnea" (50,9), "Pain" (29.6) and "Sleep disturbance" (35.2; max. score 100 = high frequency) were most often found. Further adjuvant therapy (at least 6 months after completion) in 11 patients and the type of pneumonectomy (right-side: n = 11, left-side: n = 25) did not result in additional significant reduction of lung function and of single quality of life dimensions, except that "Dyspnea" in patients with right-sided pneumonectomy (66.7) was significantly more frequent than in patients with left sided pneumonectomy (44.0). CONCLUSIONS: In this study the restricted quality of life after pneumonectomy was mainly caused by reduction of cardiopulmonary function (reduced lung function by loss of parenchyma). Adjuvant therapy reduced neither lung function nor quality of life. Compared to quality of life measurements after lung resection [17] quality of life after pneumonectomy didn't worse. PMID- 9334091 TI - [Treatment of recurrent traumatic shoulder dislocations with coracoid transfer- Latarjet-Bristow operation]. AB - We reviewed 31 patients with recurrent traumatic anterior shoulder dislocations, being operated with the Latarjet-Bristow procedure consecutively between March 1991 and March 1994. The average follow-up time was 31.2 months. According to the Rowe-score results were estimated as excellent in 45.1% and as good in 38.7%. New shoulder dislocations occurred in one case, resulting in a new shoulder dislocation rate of 3.2 per cent. Deficits in the external rotation of more than 5 degrees compared with the not operated side were seen in 15 patients. In two patients 7 respectively 9 months after the operation loosenings of the screws happened, but were without any effect on the functional outcome of the operation. The Latarjet-Bristow procedure proved to be a very safe method with a low redislocation rate, few complications and good functional results. Because of limitations in external rotation, however, this method should only be performed in cases in which an anatomic labrum reconstruction is hardly possible. PMID- 9334093 TI - [Prognostic factors in mesenteric infarct]. AB - Between 1979 and 1995 we operated 141 (80 female/61 male) patients with acute mesenteric ischemia (AMI) in our department with a median age of 71.5 years. We found 107 arterial occlusions of the intestinal arteries, 16 patients with splanchnic vein thrombosis and 18 non-occlusive-AMI. We performed 46 bowel resections, 24 vascular interventions, 11 combinations of both and in 60 cases laparotomy alone. The mortality rate was 70.9% (75% in over 70 years old patients and 65% in under 70 years old patients). Acute mesenteric ischemia remains a disease with a high mortality between 60 and 80%. Prognostic factors include the time interval until surgical intervention, elevated WBC and serum-lactate level. The mortality rate is higher in elderly patients than in younger patients which is mainly due to delayed surgery in the elderly group. If surgery is performed early the survival rate increases independent of age. The most important prognostic factor and the only factor that can be influenced by the surgeon is the time interval between onset of symptoms and surgery. Therefore angiography or laparotomy should be performed as early as possible in cases of suspected AMI. PMID- 9334094 TI - [Experiences with surgical therapy of hepatic echinococcosis]. AB - Between January 1984 and December 1990, 56 patients with hydatid liver disease were treated surgically at our Department. Diagnosis was made by using clinical criteria, serology and imaging techniques. Most frequent clinical symptom was abdominal pain or local discomfort (38 patients, 68%). Plain X-ray of the abdomen was helpful in 20 patients (36%), liver ultrasound in 53 (93%) and computerised tumorgraphy in 56 patients, (100%). The immunoelectrophoresis test of "arc 5" was sensitive in 51 patients (91%). Thirty patients (53%) underwent partial resection and omentoplasty, 17 patients (30%) underwent external drainage, two cystic resection (3%), one left lateral lobectomy (2%) and six (11%) underwent omentoplasty and T-tube insertion. Fatal complications did not occur. Four patients developed hepatic abscess (7%), three wound infection (5%), one bowel obstruction (2%) and in five instances (8%) drainage was maintained for more than three months. Of the 49 patients available for follow-up (87%), three (6%) developed recurrent disease. PMID- 9334095 TI - [Experiences with a vent button system in endoscopic and surgical gastrostomy]. AB - INTRODUCTION: In all variants of surgical or endoscopic gastrostomies, the abdominal wall is penetrated by a catheter. This reduces life-quality and often induces complications in gastrostomies. METHOD: In a prospective trial, an anti refluxive replacement button was applied as a catheter-substitute in 45 patients after conventional, laparoscopic and percutaneous-endoscopic gastrostomies. RESULTS: In 1 of 45 patients the button was falsely inserted, only two patients experienced minor local infections of the stoma. No further major complications were noted. The average usage-time is 6.3 +/- 4.9 [1-17] months, the rate of complications in 100 days is 0.06. CONCLUSIONS: The gastrostomy replacement button is safely applicable and improves clearly the comfort of gastrostomies with a low complication rate. PMID- 9334096 TI - [Stomach volume reduction balloon for weight loss: what is the justification for this controversial method?]. AB - Overweight and extreme morbid obesity are a common problem in our society. There are many therapeutic options currently available, but none of them can be considered as being ideal. One of these therapeutic interventions is the implantation of an intragastric balloon under endoscopic guidance. PURPOSE: Investigation of the effect of intragastric balloon Implantation on weight reduction. PATIENTS AND METHODS: In a prospective (and in part blinded) study 4 groups of patients (n = 10 each) were evaluated. In group I and II an intragastric balloon was implanted under endoscopic guidance. Group II had an intensive additional therapy by a dietition. Group I was advised to restrict themselves to a 1500 kcal diet. Group III did receive a sham implantation and the same additional therapy as group II. Group IV served as a control group and did not receive any type of treatment except the advice to restrict themselves to a diet. All balloons were explanted after 6 months. The overall follow-up rate was 18 months with a regular control of patients body weight. RESULTS: Patient inclusion criteria were comparable in all 4 groups. After 6 and 12 months a marked weight reduction was observed in group I and II when compared with group III and IV. This weight reduction was stable over time and reached statistical significance after 18 months. Additionally the results of group II (intensive additional therapy by a dietition) were better than in group I. Patients of group IV gained weight. No major complications due to the im- and explantation of the balloon were observed. CONCLUSIONS: The application of an intragastric balloon seems to be effective with regard of weight loss. The patient gets the chance to learn a more effective diet regimen which in our study population had a long lasting effect. Implantation of an intragastric balloon seems to be effective for a selective and highly motivated subpopulation of obese patients. PMID- 9334097 TI - [A new model for economic studies of therapies exemplified by postoperative parenteral nutrition]. AB - In this economic evaluation we compared the costs of a new therapeutic system (two-chamber bag) in total parenteral nutrition (TPN) with the comparative standard therapeutic systems (multiple-bottle system in intensive care patients followed by a combination solution (glass bottles) on the ward). In the model, standard treatment algorithms of a 10-day course TPN for patients after major gastric surgery were specified for both application systems, the two-chamber bag and the comparative system. Based on the standard treatment pattern, the resource utilisation (manpower services, medical needs, material) was assessed. In a base case analysis the types and amounts of resources were valued using salaries, prices and tariffs to assess the costs. The costs per day and per case of the therapeutic systems were compared. Sensitivity analyses were carried out to validate the cost-estimates. The total costs per patient of the two-chamber bag amounted to DM 2324.41, which was substantially less than the DM 2728.99 cost of the comparative system. The average daily costs for the two-chamber bag system were 12% to 23% lower than for the comparative system. The results were shown to be valid for the whole range of tested parameters. This model makes it possible to obtain an economic evaluation of various therapeutic modalities without undertaking a prospective randomized study with the attendant high time and cost requirements. PMID- 9334098 TI - [Thromboembolic complications in neurosurgical patients]. AB - This study was aimed at the analysis of thromboembolic risk factors in a large and representative group of adult neurosurgical patients. Using an open retrospectively designed mode of evaluation, 3162 inpatients treated in 5 years were investigated. Thromboembolism was diagnosed clinically and confirmed be ultrasound and phlebographic tests. Thromboembolism prophylaxis included subcutaneous injection of low-molecular-weight heparin and supporting physical therapy. In the whole patient population, 2.6% deep venous thromboses and 0.7% fulminant pulmonary embolisms were diagnosed. Risk factors such as malignant disease, ischemic disease of the heart, history of thromboembolism, age over 60 years, varicosis, and immobilisation rendered patients especially prone to thromboembolic events, increasing 2- to 3-fold their respective risk and reaching for some factors the level of statistical significance. Low-molecular-weight heparin application reduced the rate of thromboembolic complications, as compared to the group without heparin prophylaxis. Wound revision because of hematoma formation was necessary in 1.1% of all cases, and heparin did not induce an increase in the rate of wound hematoma formation. We conclude that low-molecular weight heparin prophylaxis is able to safely reduce the rate of thromboembolic events in neurosurgical patients without producing additional drug-related surgical complications. PMID- 9334100 TI - [Continuous "high flux" dialysis: an effective renal replacement therapy for intensive care patients]. AB - Currently available replacement therapies for the treatment of acute renal failure are reviewed. Particular interest is focused on their application in intensive care, especially in septic patients. Underlying principles and mechanisms of action are explained. The continuous "High flux-dialysis" is shown to be a particularly effective form of renal replacement therapy. PMID- 9334099 TI - [Preoperative aspirin therapy--a contraindication for kidney transplantation?]. AB - INTRODUCTION: The influence of ASA treatment on haemostasis is still debated. There are doubts in case of emergency operations, especially in transplant patients, concerning dosage and change of medication. METHODS: Our results are based on an analysis of the therapeutical behavior in German transplantation centers. The question of transplant suitability, dosage and haemostatic effects will be discussed in comparison to the literature. RESULTS: 85.7% of the transplantation centers perform the operation despite of and under ASA treatment. In these cases an increased bleeding tendency is tolerated and observed in 33.3% of the centers. An increased mortality has not been reported. Most of the transplantation centers accept a dosage of 100 mg ASA daily. Only 14.3% of the transplantation centers refuse the operation in ASA treated patients. CONCLUSIONS: Correctly indicated ASA treatment (cardiac arrhythmia, patients with embolism or thrombosis) isn't a contraindication for renal transplantation (daily dosage up to 100 mg). In elective cases however, a preoperative change of the medication is recommended, e.g. the use of heparin instead of ASA. PMID- 9334101 TI - [Medical applications of special fibers]. PMID- 9334102 TI - [Pathogenesis of puncture-site metastases after laparoscopy]. AB - The major factor underlying the seeding of tumor cells during laparoscopy are mechanical, with CO2 playing only a secondary role. The peritoneal wound is of great importance, especially in advanced tumor stages, when cells are present within the abdominal cavity. Most reported port-site metastases were found within the extraction port when no protective measures were taken. Gasless laparoscopy is no solution to the problem, since numerous port-site metastases have been described after thoracoscopy, during which no CO2 is used. The surgeon's role in the seeding of tumor cells is based on tumor perforation, excessive manipulation and replacement of trocars. This presumably explains the large differences (0 and 21%) in the reported incidence of port-site metastases. Prospective studies now show that it is possible to keep the incidence of abdominal wall metastases to about 1%-which is comparable with that seen in open surgery-by the use of a meticulous operating technique and preventive measures. PMID- 9334103 TI - [Laparoscopic cholecystectomy--effect of position changes and CO2 pneumoperitoneum on hemodynamic, respiratory and endocrinologic parameters]. AB - The effect of laparoscopic cholecystectomy on cardiopulmonary and endocrinological parameters results from various factors such as increased intraabdominal pressure (IAP), CO2, and the positioning. However, positioning has not yet been regarded. Reliable examination of the individual influencing factors requires standardized anesthesiological procedure and constant IAP. Presently, the effect of positioning is observed separately from those effects caused by the pneumoperitoneum with CO2 (PP) under standardized conditions. METHODS: 40 patients with no history of cardiopulmonary disease were analyzed. Preoperative medication, induction and management of general anesthesia, positioning of the patient and IAP (12 mmHg) were standardized. Hemodynamic, respiratory and endocrinological parameters were determined with the patient in a supine position and in the position typical for the procedure (15 degrees head-down and 10 degrees slant to the left), each with and without PP. Heart rate (ECG), endexpiratory pCO2 (peECO2), invasive blood pressure (radial art.), central venous pressure, partial arterial O2 saturation (psaO2), and ventilation pressures (peak, plateau) were monitored throughout anesthesia. The parameters pH, pCO2, BE, HCO3-, COHb, vasopressin, lactate, and ammonia were analysed in arterial and venous blood samples at predetermined set points: base line, 10 min after CO2 insufflation, 10 min after desufflation, and 1 h after extubation (cf. table 1). Statistical analysis was performed using the Wilcoxon-test with p < or = 0.05 considered statistically significant. RESULTS: Insufflation of CO2 lead to a 12% increase of heart rate in supine position and to even 18% in the position required for surgery. Same significant changes were observed for arterial blood pressure (21 or respectively 28%). Central venous pressure increased by more than 200% after CO2 insufflation. Endexpiratory pCO2 increased by 2.4 mmHg after CO2 insufflation in the supine position and by 5 mmHg in the surgical position. Ventilation pressures increased significantly by 16%. Analysis of the effect of PP on blood gases showed that pH decreased from 7.47 to 7.43, and arterial pCO2 increased by 5.1 mmHg to 38.7 mmHg and increased further after desufflation to values of up to 43.9 mmHg. Arterial pO2 decreased steadily (18% after insufflation). Vasopressin plasma levels increased exponentially from 3.03 to maximal values of 104.45 pg/ml. Ammonia and lactate showed the expected, nearly identical course. Lactate increased within the clinically and methodically irrelevative range, from 1.12 to 1.159 mmol/l. Ammonia decreased by 29%. CONCLUSIONS: The observed changes, i.e. heart rate, central venous pressure, and arterial blood pressure are caused and altered by CO2 insufflation and the various positioning of patients. The increased vasopressin concentration more than likely contributes to these changes. The query whether the position of the patient also causes a change in respiratory parameters and blood gas analysis cannot be differentiated except for the end-tidal pCO2. Inspite of the observed changes no cardiopulmonary complications occurred in this patient group. Therefore, it seems possible to omit invasive monitoring in cardiopulmonary healthy patients. In patients with concomitant history of cardiopulmonary disease, however, deteriorations due to laparoscopy should be thoroughly taken into consideration and studied further. PMID- 9334104 TI - [Controlling complications in laparoscopic cholecystectomy: diffuse parenchyma hemorrhage in the liver parenchyma]. AB - The inflammatory pericholecystic reaction to acute or subacute cholecystitis results in the involvement in the inflammatory process of connective tissue within the liver bed, with subsequent neovascularization. The inflamed wall of the gallbladder and the surrounding connective tissue also involved in the inflammatory process become fused together thus preventing dissection in this plane. As a result, the gallbladder affected by acute cholecystitis frequently has to be dissected directly out of the liver parenchyma. The resulting diffuse parenchymal bleeding proves difficult to control by cauterization. In addition, there is a danger of postoperative bile leakage occurring. Today, the use of fibrin sealing is accepted practice in the treatment of oozing haemorrhage from the resection surface of the liver following resective surgery, and for the prevention of postoperative biliary fistulae. Using special application systems, the two-component fibrin sealing can now also be employed under video-endoscopic control. Through direct application of the adhesive to the parenchyma in the liver bed using a flexible catheter, diffuse oozing bleeds can be effectively arrested. In addition, coagulation-related parenchymal necroses associated with the development of biliary fistulae can be avoided. The technique of video endoscopic controlled fibrin sealing is an important method of preventing and controlling complications arising during video-endoscopic surgery. PMID- 9334105 TI - [Aneurysms of the iliac artery in fibromuscular dysplasia as differential diagnostic consideration in acute lower abdominal pain]. AB - A 48 year old woman was admitted to our department with sudden onset of pain in the left lower abdominal quadrant and the initial diagnosis of diverticulitis or left sided adnexitis was made. On physical examination she was found to have pain and localized peritonism in the left lower abdominal quadrant. Ischemic signs in the same sided lower limb could not be observed. Further diagnostics (abdominal ultrasonography, transvaginal ultrasonography, angiography and computed tomography) showed a covered perforated aneurysm in the left common or internal iliac artery as the reason for the abdominal pain. Coincidental focal aneurysms were seen in the middle segment of both renal arteries and also in the right common iliac artery. The patient was treated with an aorto-iliac-bypass (Impra prosthetic graft). Histological examination showed features of fibromuscular dysplasia. We are reporting this case due to the unusual localisation, the atypical symptoms and the difficult diagnostics. PMID- 9334106 TI - [Nonadrenergic noncholinergic regulation of gallstone containing and gallstone free human gallbladders]. AB - This study is to determine the role of nitric oxide (NO), as primary neurotransmitter of the non-adrenergic noncholinergic (NANC) innervation, of stone-diseased and stone-free human gallbladders. Human gallbladder muscle strips were mounted in modified Krebs-Henseleit-solution with atropine (1 microM), guanethidine sulf. (5 microM) and aerated with Carbogen. Electrical field stimulation (EFS, 70V, 0.5 ms, 100 pulses) was used at frequencies of 1, 3, 10 Hz to activate NANC nerves, L-omega-nitro-L-arginine (L-NNA, 100 microM), L-arginine (L-ARG, 120 microM) was used to manipulate the NO-synthase. Gallbladder slices of 3 microns were stained by means of APAAP-method (alkaline phosphatase anti alkaline phosphatase) for histological examination. In the control group (basal tone = 8.94 +/- 1.17 mN) EFS caused a frequency dependent reduction of basal tone (1 Hz = 5.73 +/- 0.81 mN; 3 Hz = 5.18 +/- 0.65 mN; 10 Hz = 4.63 +/- 0.49 mN). Incubation with L-NNA increased the tone (7.63 +/- 0.76 mN). Contractor group (basal tone = 7.79 +/- 0.93 mN) reacted like the control group but frequency independent and additionally with spontaneous phasic contractions. In the non contractor group (basal tone 4.13 +/- 0.65 mN) EFS only decreased the frequency of spontaneous phasic contractions. L-NNA caused an increase in tone (5.97 +/- 0.84 mN) and frequency, L-Arginine significantly reversed this effect. HISTOLOGY: Contractor group showed wrinkled mucosal membrane and mild grade of inflammation. Shallow mucosa, necrosis and high grade of inflammation were found in the non contractor group. CONCLUSIONS: 1. In vitro, NANC-relaxation of human gallbladder is NO dependent. 2. Motility of stone-diseased gallbladders is modulated by NO and seems to depend on the degree of scarrification. PMID- 9334107 TI - [Diagnosis and therapy of adrenal gland tumors]. PMID- 9334108 TI - [Molecular biology of incidentally diagnosed adrenal gland space-occupying lesion]. AB - The incidentally detected adrenal mass is with a prevalence of 1% in the general population the most common pathological process of the adrenal gland. In more than 85% of the cases it is caused by benign adenomas of the adrenal cortex. These tumors have a monoclonal composition and are, therefore, caused by oncogenic mutations with consecutive clonal expansion of this cell clone. In contrast to adrenocortical carcinoma, in which mutations of the IGF II gene locus and the p53 tumor suppressor gene has been found, the oncogenes involved in the tumorigenesis of adrenal adenomas have not been identified yet. However, opposite to other endocrine tumors the receptor-cAMP-proteinkinase A signaling pathway is not involved in the pathogenesis of these tumors. Insulin may be an important growth factor of incidentally detected adrenal tumors. Heterozygote 21 hydroxylase deficiency, however, does not seem to play a major role in the tumorigenesis of adrenal incidentalomas. PMID- 9334109 TI - [Imaging methods in diagnosis of pheochromocytoma]. AB - In the radiological evaluation of adrenal masses today computed tomography (CT) has the highest sensitivity of all imaging modalities for tumor detection. In addition, CT is able to assess potential concomitant processes in the surrounding tissues. Magnetic resonance imaging (MRI) is the method of choice, when adrenal masses cannot be differentiated by CT. When chemical-shift imaging, fat suppression, and dynamic contrast-enhanced sequences are used for tissue characterization, pheochromocytomas can be distinguished from adenomas. Due to the multiplanar imaging ability MRI is optimal for the preoperative work-up of the anatomical situation in patients with adrenal masses. Extraadrenal paragangliomas can be assessed with MRI, too. Ultrasound is of minor relevance in the assessment of pheochromocytomas and other adrenal masses, as is angiography, which is only carried out in special cases to evaluate the vascular supply. PMID- 9334110 TI - [Nuclear medicine diagnosis of pheochromocytoma]. AB - The synthesis of radioiodinated meta-iodobenzylguanidine (MIBG), a physiologic norepinephrine analog, was first accomplished by a group from the University of Michigan. Since the first report of the detection of pheochromocytoma with MIBG in humans, there have been several studies about the role of this agent in the diagnosis of pheochromocytoma and neuroblastoma. With a sensitivity of about 90% and a specificity of nearly 100% MIBG has been shown to be a reliable diagnostic tool for the detection and follow-up of pheochromocytoma. PMID- 9334112 TI - [Premedication in pheochromocytoma]. AB - Pheochromocytomas are active, catecholamine-producing tumours derived from chromaffine tissue, causing arterial hypertension. The therapeutical aim is the surgical removal of the tumour. Blockade of the alpha-adrenergic receptors during the preoperative phase is necessary to prevent cardiovascular complications. Phenoxybenzamine is the commonly used alpha-adrenergic blocking agent. Therapy begins with low doses and is increased stepwise up to 250 mg daily. The optimal duration of preoperative therapy is 10 to 14 days. Efficiency of therapy should be judged by reduction of symptoms, a stabilisation of arterial blood pressure, and the presence of light orthostatic hypotension. Swelling of nasal mucosa indicates a successful blockade of the alpha-adrenergic receptors. Persistent tachyarrhythmia is an indication to apply beta-adrenal receptor blockers, but only after alpha-blockade. Reexpansion of blood volume has to be adjusted according to hemodynamic monitoring data. With an adequate preoperative management, mortality of patients with pheochromocytomas has been reduced to less than 1 percent. Postoperatively, most patients become normotensive and other symptoms of excessive catecholamine-production disappear. In cases of long persisting and fixed hypertension paroxysmal crises of high blood pressure can be avoided. PMID- 9334111 TI - [Clinical and endocrine diagnosis of pheochromocytoma]. AB - Pheochromocytomas are catecholamine-producing tumors, representing one of the most important causes of secondary hypertension. The classification of these tumors considers both sporadic and familial forms, intra- and extraadrenal localization as well as the dignity. Familial pheochromocytomas are primarily seen under the conditions of multiple endocrine neoplasia, von Hippel-Lindau disease or neurofibromatosis type 1. The list of clinical symptoms includes hypertension, which can be both continuous or intermittent, headache, tachycardia and sweating. It is most important to standardize the pre-analytical procedures, i.e. control for sampling conditions and adequate choice of parameters in plasma or urine. For screening sensitive methods will be employed (free catecholamines in 24h-urine) and for confirmation of the diagnosis, specific procedures are performed (Clonidine test, MIBG-scintigraphy). The endocrinological and biochemical procedures are completed by molecular genetic techniques in familial pheochromocytoma. PMID- 9334113 TI - [Anesthetic management in pheochromocytoma]. AB - Consequent preoperative alpha-receptor blockade, as well as improved surgical technique, anaesthetic management and monitoring have reduced perioperative mortality below 3%. Nevertheless, serious problems are still to be expected during the resection of "complicated" pheochromocytoma. "Complicated" in this context means one or more of the following conditions: missing preparation, severe technical surgical difficulties (e.g. location in the liver), concomitant disorders (e.g. coronary arterial disease), pregnancy and/ or great consequential damages (e.g. cardiomyopathia). Therapy of hemodynamic crisis during "complicated" resection of pheochromocytoma requires differentiated pharmacological combinations considering concomitant disorders. For instance multiple various combinations of phenoxibenzamine, natriumuitropussid, adenosine, magnesiumsulphate, esmolol, diltiazem have been used successfully in concomitant coronary art disease. Absolute preferences are not established. Similarly in the anaesthetic management, a determination of a preferred procedure is still missing. All anaesthetic possibilities should be used carefully directed with two aims: first minimise indirect release of catecholamines through effective vegetative damping and prevention of stress; second to support therapy of hemodynamic crises through ingenious use of effect and side effects of the anaesthetic drugs. Experiences and excellent cooperation between surgery, endocrinology and anaesthesiology are essential for successful prevention of perioperative complications. PMID- 9334114 TI - [Surgical concept in sporadic and familial pheochromocytoma]. AB - A retrospective analysis of 61 patients operated upon sporadic and familial pheochromocytomas demonstrates the changes in diagnosis and therapy. Familial pheochromocytomas make up for 28% of all patients and in 13% tumor malignancy was diagnosed. Patients had suffered from endocrine crises in 21% preoperatively but only in 3% intra- and postoperatively. CT was confirmed to visualize more tumors than US or MIBG (> 90%) preoperatively. Operative strategy was based on tumor size and tumor-localization but increasing importance of endoscopic procedures is conceded. Postoperative complications are rare (infection 8%; Addison's crisis 5%) and life threatening complications (bleeding, post op death) as well as tumor persistence or recurrence almost exclusively occur in patients with malignant disease. Altogether the main changes are the early diagnosis of especially familial disease and the use of endoscopic procedures in the treatment of patients with small benign pheochromocytomas. PMID- 9334115 TI - [Conventional operation for therapy of incidentaloma]. AB - AIM: The increasing detection rate of incidentalomas imposes not only the question about the necessity of treatment but also with introduction of laparoscopic adrenalectomy the question about operative strategy. Own patients and literature are analysed and discussed. PATIENTS: Between 1985 and 1996 203 patients have been operated on for adrenal disease including 21 patients with incidentaloma. RESULTS: Biochemical analysis revealed latent hormonal activity in 9 patients. All patients were operated conventionally. Pathohistology demonstrated 1 adrenal carcinoma, 9 adrenocortical adenomas, 2 myelolipomas, 2 adrenal cysts, 2 nodular hyperplasias and 5 pheochromocytomas. Postoperative morbidity was 5% and mortality 0%. CONCLUSION: Small and hormonally latent active tumors can be operated laparoscopically or conventionally via dorsal approach. In case of suspected malignancy conventional operation is the more reliable procedure. PMID- 9334116 TI - [Endoscopic therapy of incidentaloma]. AB - Since 1992 endoscopic surgical techniques have gained importance in adrenal gland surgery. In this review the actual laparoscopic and retroperitoneoscopic techniques and results-taking the own case material into consideration-are represented. The laparoscopic, transperitoneal adrenalectomy shows a low rate of complications (12%). Disadvantages of the transperitoncal approach are the risk of an intraabdominal injury and the problems caused by adhesions after abdominal operations. The retroperitoneoscopic access to the adrenal gland avoids disadvantages of the transperitoneal approach. Concerning the rising frequency of accidentally diagnosed tumors of the adrenal gland the retroperitoneoscopic approach represents a technique which allows a removal of these tumors up to a size 6 centimeters with low morbidity. In case of large tumors of the adrenal gland (size > 6 centimeters) suspected for malignancy we see the indication for the transperitoneal approach. PMID- 9334117 TI - [Surgery of primary unilateral adrenal gland tumors--results of 154 patients]. AB - Surgical resection of adrenal neoplasias with endocrine activity is principally indicated. In adrenal neoplasias without endocrine activity, surgical removal is indicated in relation to tumor size. Surgical access and extent of resection are the major problems related to adrenal surgery. From 1980 to 1996, in 154 patients (62 m, 92 f) primary and unilateral adrenal tumors (139 benign, 15 malign) were resected. 93 resections were performed transperitoneally, 13 extraperitoneally, and 48 retroperitoneoscopically. Subtotal adrenal resections were performed in 23 benign tumors smaller than 4 cm. Perioperative lethality was 0%, morbidity was 31.8%. Malignancy was correlated to tumor size: In 114 tumors smaller than 5 cm, no malign neoplasia was found, whereas in 40 tumors larger than 5 cm, 15 specimen were malign. Operating time of the retroperitoneoscopic method was significantly longer than of open procedures (p < 0.05). Postoperative analgotic medication was significantly reduced after endoscopic surgery compared to transperitoneal or extraperitoneal surgery (p < 0.0001). No tumor recurrences occurred after subtotal adrenal resections (mean follow up: 5.7 [1.3 years]). In patients with adrenal carcinomas, 5-year-survival was approximately 15%. In adrenal neoplasias smaller than 5 cm, malignancy is extremely rare. Therefore, less aggressive surgery with a lower morbidity (extraperitoneal approach) and reduced postoperative pain (retroperitoneoscopic approach) including function preserving resection is indicated in these lesions. Due to the high incidence of malignancy, adrenal tumors larger than 5 cm should principally be treated by conventional transperitoneal surgery. PMID- 9334118 TI - [Adrenal gland incidentaloma is not a "time bomb"--arguments for follow-up control]. AB - Adrenal masses are nowadays more and more diagnosed incidentally. In the vast majority of cases these "incidentalomas" are benign adrenocortical adenomas which do not compel operative therapy. In about 12% of patients an endocrine activity is detected. With Increasing tumor size the risk of malignancy increases. This article summarizes recommendations, only to allow patients with endocrine activity or an increased risk of malignancy to be selected for surgery. A small uncertainty will always remain, but may be minimized by an individually tailored follow-up. PMID- 9334119 TI - [Therapeutic possibilities in metastatic pheochromocytoma]. AB - Malignant pheochromocytomas are rare. Although 5-year survival is less than 50%, the prognosis varies. Some patients, even those with extensive metastases, have been followed up for many years. If the tumor tissue's uptake is adequate (> 5 Gy/100 mCi) the therapeutic use of 131I-meta-iodobenzylguanidin (131I-MIBG) is at present the therapy of first choice. The use of cytostatic chemotherapy should be limited to patients with rapidly progressive disease. PMID- 9334121 TI - [Prof. Dr. Lothar Heidenhain (1860-1940)--on the first successful operation of congenital diaphragmatic hernia 6 March 1902]. AB - A biographic sketch of Heidenhain is presented referring to Heidenhain's work in all parts of surgery, especially in breast, brain and lung surgery and cancer research. He was the first operating successfully an inborn diaphragmatic hernia. PMID- 9334120 TI - [Laparoscopic cholecystectomy in antegrade (prograde) technique]. AB - In the case of acute cholecystitis and chronic cholecystitis of lang standing, the inflammatory changes in Calot's triangle make the risk of damaging the bile duct during laparoscopic cholecystectomy particularly high. In view of the difficult anatomical situation in Calot's triangle, such patho-anatomical conditions when encountered during open surgery best dealt with by anterograde (prograde) dissection of the gallbladder beginning from the fundus and proceeding towards the neck of the gallbladder. Since this approach is considered to be safer and less risky, it should also be adopted during laparoscopic surgery. The advantage is to be seen in the initial dissection far removed from the bile ducts. Despite pronounced inflammatory changes and initial preservation of the cystic artery, the use of modern technologies permit a virtually bloodless procedure. Nevertheless, the difficult anatomical situation makes a high level of readiness to conver mandatory. Should there be any doubt, the open surgical approach is the safer modality. PMID- 9334122 TI - [Comment on the contribution by T. Hohaus, G. Hellmich, K. Ludwig, "Unexpected gallbladder carcinoma--a report of experiences after 1,200 laparoscopic cholecystectomies]. PMID- 9334124 TI - [Intensive care in increased intracranial pressure. II: General intensive care treatment of patients with increased intracranial pressure. Ventilation. Intensive Care Committee of the German Society of Anesthesia and Intensive care]. PMID- 9334125 TI - [Fluid management of patients with craniocerebral trauma]. PMID- 9334123 TI - [A simple technique for reconstruction of anterior and middle cranial base after palliative tumor resection]. AB - Large defects of the anterior and middle skull base resulting from extensive tumour resection should be reconstructed. Reconstruction should not only include watertight closure of the dura but also osseous reconstruction. Numerous alloplastic and autoplastic grafts have been used for this purpose. Following curative tumour resection autoplastic material is used at our institution. Autoplastic bone/however requires additional procedures, adds additional operating time, and often requires surgical recontouring because of its rigidity. Therefore we have used a simplified technique using autologous grafts for dural repair in combination with a Vitallium mesh for osseous reconstruction after palliative tumour surgery. So far this technique has been used in 10 patients with different malignancies of the anterior and middle skull base. No complications with the implant was observed. There was no operative mortality and morbidity. One postoperative meningitis was cured by antibiotics. Compared to autogenous bone the technique saved operating time, had more intraoperative flexibility, and a low complication rate with good cosmetic results. PMID- 9334126 TI - [Nutrition for patients with craniocerebral trauma]. PMID- 9334127 TI - [Cerebrovascular effects of analgosedation]. PMID- 9334128 TI - [Critical care of patients with increased intracranial pressure]. PMID- 9334129 TI - [Reports by the German Interdisciplinary Group of Intensive Care and Emergency Medicine. New recommendations for medical qualifications in transport of intensive care patients]. PMID- 9334130 TI - [A new Bordetella species in sheep?]. AB - Occasionally the occurrence of isolates of the genus Bordetella has been reported, with unclear assignment to one of the known species in sheep from New Zealand and Great Britain. In this study we describe the isolation of strains belonging to the genus Bordetella in a flock of sheep from Germany. These isolates were characterized biochemically by serological tests and whole cell fatty acid analysis. Our isolates could be divided into three subgroups by their differential growth on MacConcey- and Tyrosine agar. A clear assignment to one of the known Bordetella species was not possible. PMID- 9334131 TI - Fine needle aspiration biopsy of metastatic malignant melanoma with "rhabdoid" features. Frequency, cytologic features, pitfalls and ancillary studies. AB - OBJECTIVE: To characterize the cytopathology of metastatic malignant melanoma (MM) with "rhabdoid" features, a recently described, rare morphologic variant of MM that can be incorrectly diagnosed in fine needle aspiration (FNA) biopsy. STUDY DESIGN: A retrospective review of all FNA biopsy material with the diagnosis of metastatic MM was performed at two institutions. Only cases with a predominant composition of cells that met criteria defined as "rhabdoid" morphology were selected for study. The cytomorphologic features, immunocytochemistry and clinical features of these cases were reviewed. RESULTS: Of 88 FNA cases previously diagnosed as metastatic MM, 4 (4.6%) had a predominance of cells with rhabdoid features. These cases consisted of scattered atypical cells having enlarged, eccentrically placed nuclei; prominent nucleoli; and a moderate amount of cytoplasm possessing round, globular inclusions in Papanicolaou- and Diff-Quik-stained smears. Immunochemistry showed strong S-100, HMB-45 and vimentin staining in two of four cases. CONCLUSIONS: Metastatic MM may present in FNA biopsy as a poorly differentiated malignancy with rhabdoid features, potentially leading to an incorrect cytologic diagnosis. MM must be considered when evaluating neoplasms with a rhabdoid phenotype. Correlation of the cytologic finding with the clinical history and immunohistochemical studies can help in diagnosing this morphologic variant. PMID- 9334132 TI - Natural family planning and reproductive health awareness: expanding options and improving health. Symposium proceedings. Washington, D.C., April 2-5, 1997. PMID- 9334133 TI - Tibor Kobulej (1921-1997). PMID- 9334134 TI - It's not the principle of the thing. PMID- 9334135 TI - Appropriate use of hysterectomy. PMID- 9334136 TI - [Comparative ontogenic analysis of the epithelium of the non-glandular stomach compartments of merino sheep]. AB - A total of 74 embryos and fetuses were used in a comparative analysis of the epithelium of the non-glandular stomach compartments of merino sheep during development. The mechanical protection showed by the tegumentary epithelium in the superficial layers of the rumen, reticulum and omasum is supported by a buffer system of neutral mucopolysaccharides secreted by the deeper strata. Neutral mucopolysaccharides first appeared in epithelial cells at 46 days of fetal life. Acid mucopolysaccharides, mucins, and mucoid compounds were not detected. Growth curves and formulas were constructed for the epithelial layers. PMID- 9334137 TI - [Changes in the shape and growth of motor endplates of mm. fibularis longus, semitendinosus and longissimus in pigs]. AB - Age-related changes in motor endplates were studied in the fibularis longus, semitendinosus and longissimus muscles in pigs. They develop from small, compact AChE-active figures in fetuses to typical myoneuronal synapses with a branched subsynaptical structure of AChE-active units. High correlations were found between size of endplates and both myofibre diameter and body mass. Growth analyses with respect to age, as well as allometrical approaches, showed that endplate growth slightly precedes that of myofibres, with both having similar growth patterns. The morphological changes, however, imply that the increment of synaptical area could lag behind that of myofibres. PMID- 9334139 TI - Neuropsychiatric Manifestations of Systematic Lupus Erythematosus. Proceedings of a conference. New York City, New York, September 27-30, 1996. PMID- 9334138 TI - Nurses can choose less stressful lives. PMID- 9334140 TI - Outcomes '97. Proceedings and abstracts of conference on cardiac and vascular surgery: neurobehavioral assessment, physiological monitoring and cerebral protective strategies. Key West, Florida, April 24-27, 1997. PMID- 9334141 TI - [Immunomodulating, antioxidant and hepatoprotective action of the phytopreparation lochein]. AB - Lochein is a water soluble extract from Siberian plants of the family Chenopodiaceae. It was shown to stimulate the immune response to the T-dependent antigen in healthy animals and in animals exposed to hepatotropic poison. Lochein induced secretion of immunostimulating factors by the glass adherent and nonadherent spleen cells. Supernatants of the glass adherent spleen cells contained compounds with antioxidant and hepatoprotective properties. PMID- 9334143 TI - [Pefloxacin mesylate--clinical effectiveness in various forms of infectious inflammatory diseases]. AB - Pefloxacin mesylate is a broad spectrum fluoroquinolone active against pathogens with multiple resistance. Due to its high therapeutic efficacy (at last 90 per cent) and good tolerance pefloxacin mesylate is considered as the most promising drug in the therapy of severe infections and infections difficult for the treatment such as wound infection, meningitis and particularly perilous infections. Prolonged pharmacokinetics and high bioavailability of the drug provided its administration in the treatment of in- and outpatients with severe infection twice a day in a daily dose of 0.8 to 1.2 g. Pefloxacin proved to be a drug of choice in the treatment of infection due to intracellular pathogens. PMID- 9334142 TI - [Effectiveness of cefotetan in clinical practice]. AB - Forty five patients at the age of 15 to 84 years with signs of infection requiring active antibacterial therapy were treated with cefotetan. In the majority of the patients pulmonary affections such as double pneumonia, pleurisy or bronchopneumonia were stated. In some patients bronchopulmonary pathological processes were associated with pancreatitis, cholecystitis or other diseases of the gastrointestinal tract. A separate group included patients with diseases of the small pelvis organs (pelvioperitonitis, metroendometritis or prostatitis) and diseases of the urogenital system (pyelonephritis) arachnoiditis. In all the patients except for one with bronchopneumonia at the background of chronic myeloleukemia and agranulocytosis the results of the treatment were good and satisfactory. Cefotetan proved to be efficient in the treatment of purulent affections of the skin and subcutaneous fat (abscesses and phlegmona), trophic disturbances at the background of pathological processes in the vessels and pyoseptic condition. Cefotetan practically had no side effects. Only in 2 patients insignificant nausea during the first 2 days of the treatment was recorded. In some patients the antibiotic intramuscular injections were painful with formation of cold infiltrates. After intravenous administration of cefotetan no adverse reactions were observed. PMID- 9334144 TI - [New cephalosporins, effective in methicillin-resistance]. PMID- 9334145 TI - [Cyclosporin in ophthalmology]. PMID- 9334146 TI - [Effect of the amide of 1-adamantanecarboxylic acid (AACA) and its combination with ribavirin on the course of experimental infection, caused by the sindbis virus in tissue culture and in mouse brain. Possible isolation of the virus mutant resistant to ribavirin and to a combination of the inhibitors]. AB - It was shown advisable to use ribavirin (a weak inhibitor of Sindbis virus reproduction) in combination with ACAA. The effect observed was of marked synergistic nature. Subcultures of the susceptible strain in the presence of ribavirin/ ACAA did not lead to the development of resistance to ACAA, ribavirin or their combination. The ACAA resistant strain preserved its susceptibility to ribavirin after serial subcultures in the presence of both the inhibitors. However, no synergistic effect of the combination under such conditions was observed. It was of principal importance that in the subcultures of the ACAA resistant strain in the presence of ribavirin its susceptibility to ACAA recovered. ACAA was efficient in inhibition of Sindbis virus reproduction in the brain of the intracerebrally infected unimbred albino mice. The maximum inhibitory effect was observed after repeated administrations of ACAA under the treatment and prophylactic regimens. Ribavirin markedly increased the ACAA protective effect. PMID- 9334147 TI - [Probiotics and functional food]. PMID- 9334148 TI - [Effect of promethazine hydrochloride (pipolphen) on the stability of R plasmid resistance in Escherichia coli]. AB - The influence of promethazin hydrochloride (pipolphen) on stability of R plasmid inheritance in Escherichia coli strains of various serogroups was studied. The strains were isolated from patients with acute intestinal infection and from healthy persons. It was shown that in subbacteriostatic concentrations (100 to 450 micrograms/ml) pipolphen promoted elimination of the R plasmids. Decreased stability of the R plasmid inheritance was not associated with the pipolphen concentration. No influence of the drug on the biochemical characteristics, antigenic properties and nutritional requirements of the plasmid-free derivatives was detected. The eliminating action of pipolphen and ethidium bromide in some strans of Escherichia coli was shown to be different. PMID- 9334149 TI - [Main steps of immunohistochemical diagnosis of human tumors]. AB - Most rational schemes of immunohistochemical diagnosis based on the minimal spectrum of antibodies as the 1st and the 2nd steps of investigation are given. The importance of joint use of immunohistochemistry and electron microscopy in histogenetic diagnosis of tumors is emphasized; many diagnostic problems can be solved by the use of both methods. PMID- 9334150 TI - [Immunohistochemical study of oncogene c-erbb-2 expression and extracellular matrix proteins in breast cancer (various mechanisms of invasion)]. AB - 55 breast carcinomas and 9 its benign conditions were studied immunohistochemically. Oncoprotein c-erbB-2 was found in 22% carcinomas and in 33 37% comedo-carcinomas. Coincidence of predominant expression of oncoprotein c erbB-2 and tenascin was observed. Maximal damage of the basal membrane in the intraductal carcinoma detected by the disturbance of type IV collagen staining was observed in cases of the highest expression of oncoprotein c-erbB-2 and tenascin. PMID- 9334151 TI - [Changes in the endocrine apparatus of the gastric mucosa in forms of cancer of varying origin]. AB - The authors studied endocrine apparatus of the mucous membrane of 53 stomachs in various forms of carcinoma. Silver impregnation and electron microscopy were used as well as routine histology and histochemistry. All the tumors were divided into endocrine-cell and non-endocrine-cell tumors (ET and NET). Cells of the diffuse endocrine system take an important part in the development of the background and pretumorous processes in the stomach mucous membrane. Endocrinocyte hyperplasia, degree I and II, of the mucous membrane of the antrum and enterolysation foci was the background for all NET. Endocrinocyte hyperplasia was more pronounced (degree II and III) in ET and spread to the fundal glands being combined with endocrinocyte dysplasia and metaplasia. These changes are assessed as precancerous for tumors with high content of endocrinocytes. PMID- 9334152 TI - [Therapeutic pathomorphism of Ewing's tumor]. AB - Pathomorphosis of Ewing's tumors after the combined treatment is studied with the use of light and electron microscopy, DNA-flow cytometry. Necrosis and apoptosis of tumor cells produced by the therapy are characterized. Therapeutic forms of tumor cells representing hyperaneuploid population delayed at the G2-level of the cell cycle are described. PMID- 9334153 TI - [Teaching on pathomorphism: past and present]. AB - The history of the pathomorphosis teaching is outlined. The study of this problem should rest on the knowledge of general and partial causes underlying pathomorphosis, peculiar features and contribution of each of these causes and their interaction. PMID- 9334154 TI - [Aminopeptidases in cells of non-Hodgkin's lymphoma of varying degrees of malignancy and in lymphogranulomatosis]. AB - 26 cases of lymphoproliferative diseases were studied: 8 cases of reactive follicular hyperplasia (RFH), 11 cases of non-Hodgkin's malignant lymphomas (NML), 7 cases of lymphogranulomatosis (LGM). Only gamma-glutamyl transpeptidase (GGT) was found in lymphoid cells of B- and T-dependent areas of lymph nodes with reactive changes as well as in tumor cells of NML and LGM. GGT activity was more pronounced in NML of high-grade malignancy (centroblast and immunoblast) as compared to lymphomas of lower grade of malignancy (lymphocytic, centroblast centrocytic and in Lennert lymphoma). GGT activity in cells of Hodgkin and Berezovsky-Sterberg in some cases of LGM was high, in others low. Significant differences in GGT activity between RFH and follicular centroblast-centrocytic lymphoma were not found. Activity of aminopeptidase M was observed in histiocytes, fibroblasts, vessels and areas of connective tissue growth. Aminopeptidase A activity was observed in vessels only. Activity of dipeptidyl(amino)peptidase IV was observed in some lymphoid cells in RFH, NML and LGM. Thus, GGT activity may be considered as a differential-diagnostic marker in separating NML of high and low degree of malignancy and this may presume a different sensitivity to the therapy. PMID- 9334155 TI - [One decade after Chernobyl: sequelae of the accident and urgent problems of radiation pathology]. AB - Medico-biological sequelae of irradiation of humans as a result of the Chernobyl accident are considered. Acute radiation sickness, thyroid carcinoma in children, chromosomal aberrations in the peripheral blood lymphocytes have an etiological link with radiation. Three problems are important for future studies: 1) collection of reliable information about the radiation, 2) assessment of dose effect relations, 3) organization of supervision. PMID- 9334156 TI - [Mucous-secreting breast cancer]. AB - 3 groups are distinguished among 219 cases of the breast carcinoma with mucus formation: one-component colloid carcinoma, combined colloid carcinoma and signet ring cell carcinoma. Morphological features and prognosis for each group are listed. One-component colloid carcinoma has the most favourable prognosis: incidence of distant metastases 15% versus 27% and 33% in combined colloid and signet ring cell carcinoma, respectively. Signet ring cell carcinoma should obviously be regarded as a variant of invasive lobular carcinoma. PMID- 9334157 TI - [Phenotypic characterization of cells and structure of the extracellular matrix in pleomorphic adenoma of the salivary glands]. AB - Phenotypic characteristics of cells and extracellular matrix (ECM) of 14 salivary gland pleomorphic adenomas were studied by indirect immunofluorescence. All cells in the epithelial structures expressed cytokeratin 8, the majority of cells produced vimentin, laminin and IV type collagen. Cells of myxoid areas expressed vimentin: type IV collagen was observed in the form of fibrillar structures. Fibrous areas were characterized by vimentin and cytokeratin 8 expression; types I, III, IV collagens and fibronectin with ED-A sequence were present in ECM. Terms "epithelial structures", "epithelial" and "myoepithelial" cells in relation to pleomorphic adenoma may be used only relatively as it is mainly represented by cells with features of both epithelial and mesenchymal phenotype. Characteristics of ECM synthesized in various structures of pleomorphic adenoma may serve an additional criterion of cell transformation. PMID- 9334158 TI - [Hyperplastic follicular response of the regional lymph nodes in cancer (immunohistochemistry, ultrastructure, prognostic importance)]. AB - 194 regional lymph nodes surgically removed because of cancer at various sites from 60 cancer patients are studied immunohistochemically and ultrastructurally. Monoclonal antibodies (MCAB) against antigens CD 1, CD 2, CD 3, CD 4, CD 8, CD 10, CD 19, CD 20, CD 30, CD 45, CD 56, BLA-36 were used for differentiating lymphoid cells. MCAB against alpha 1-antichymotrypsin, lysozyme, S100 protein, desmin, pan-cytokeratin, vimentin, laminin, collagen type IV, factor VIII; CD 35 were used as markers of non-lymphoid cells. Immunohistochemical classification of the hyperplastic follicular reaction is proposed: 1) typical B-cell follicle; 2) B-cell follicle with high content of T- and NK-cells; a) with domination of helper-inducer subpopulation, b) with domination of killer-suppressor sub population; III) T-cell follicle; IV) non-lymphoid cell follicle. It was established that type I and IV follicular reaction correlates with a low 5-year survival, while type IIb and III. hyperplasia correlates with a favourable prognosis. PMID- 9334159 TI - [Necessity for unification of the clinico-anatomic classification of tuberculosis]. AB - Critical assessment of tuberculosis classification adopted at II(XII) Congress of Phthisiatrists Association on September 7-9 1994 in Saratov as well as the section "Tuberculosis" in the International statistical classification of diseases and problems associated with health 10th reevaluation is presented. Main critical remarks are the following: organo-pathological principles of the classification, uncertainty of the definitions, ignorance of the iatrogenic conditions. PMID- 9334160 TI - [Comprehensive evaluation of the work of medical pathologists]. AB - The quantitative criteria system of complex assessment of pathologist's work is presented. It is based on the results of comparative quantitative analysis of the work made, standard and mean rates. Labor activity of pathologists was assessed by its intensity, complexity and quality with further introduction of the integral index. Such an approach improves objective assessment of pathologists' qualification and facilitates work of medical managers. PMID- 9334161 TI - [A method of improving the attachment of paraffin sections to a glass slide]. PMID- 9334162 TI - [Main principles and diagnostic criteria of "Reevaluation of the European American classification of lymphoid tumors"]. PMID- 9334163 TI - [Non-inflammatory form of neutrophil antibacterial response]. AB - According to the Mechnikov theory inflammation is the only form of the protective adaptive response of phagocytes and phagocytosis is its essence. High doses of Staphylococcus aureus (25 x 10(6) and 25 x 10(2) bacteria) in the absence of additional peritoneal damage do not produce the inflammation. The organism of the highest animals possess non-phlogogenic mechanisms of protection against bacteria when the phagocyte function is not the destruction of bacteria but their catching and transportation to the intestinal lumen. PMID- 9334165 TI - A classification of nucleotide-diphospho-sugar glycosyltransferases based on amino acid sequence similarities. PMID- 9334164 TI - Interaction of platelet-derived growth factor with thrombospondin 1. AB - Key factors that mediate vascular smooth muscle cell proliferation and migration are platelet-derived growth factor (PDGF) and thrombospondin 1 (TSP1). We now report that PDGFBB bound tightly and specifically to TSP1, that this interaction was markedly dependent on the disulphide bond arrangement in TSP1, and that binding of PDGFBB to TSP1 did not preclude PDGFBB from binding to its receptor on rat aortic vascular smooth-muscle cells. At physiologic ionic strength and pH, PDGFBB bound to Ca2+-depleted TSP1 with a dissociation constant of 11 +/- 2 nM and to Ca2+-replete TSP1 with a dissociation constant of 32 +/- 5 nM. Binding was specific, as both soluble TSP1 and unlabelled PDGFBB competed for binding of iodinated PDGFBB to immobilized TSP1, whereas other platelet alpha-granule proteins did not compete. The tertiary structure of TSP1 is regulated by intramolecular disulphide interchange; we found that catalysis of disulphide interchange in TSP1 by protein disulphide isomerase ablated the binding of PDGFBB. The interaction of PDGFBB with TSP1 was weakened by increasing salt concentration and essentially ablated at 0.65 ionic strength; it was inhibited by heparin with a half-maximal effect at 20 i.u./ml, implying that the binding was mediated largely by ionic interactions. An anti TSP1 monoclonal antibody decreased the binding of iodinated PDGFBB to PDGF receptor on rat aortic vascular smooth-muscle cells by 37 +/- 2%, whereas platelet TSP1 non-competitively inhibited binding of iodinated PDGFBB. Uncomplexed PDGFBB bound to PDGF receptor with an affinity 5 +/- 2 times that of PDGFBB-TSP1 complexes. These results suggest that TSP1 might assist in the targeting of PDGF to its receptor on vascular smooth-muscle cells. PMID- 9334166 TI - The Factor I and follistatin domain families: the return of a prodigal son. PMID- 9334168 TI - The chicken lysozyme locus as a paradigm for the complex developmental regulation of eukaryotic gene loci. PMID- 9334169 TI - Tumor necrosis factor (TNF) receptor 1 signaling downstream of TNF receptor associated factor 2. Nuclear factor kappaB (NFkappaB)-inducing kinase requirement for activation of activating protein 1 and NFkappaB but not of c-Jun N-terminal kinase/stress-activated protein kinase. AB - Like other members of the tumor necrosis factor (TNF) receptor family, p55 TNF receptor 1 (TNF-R1) lacks intrinsic signaling capacity and transduces signals by recruiting associating molecules. The TNF-R1 associated death domain protein interacts with the p55 TNF-R1 cytoplasmic domain and recruits the Fas-associated death domain protein (which directly activates the apoptotic proteases), the protein kinase receptor interacting protein, and TNF receptor-associated factor 2 (TRAF2). TRAF2 has previously been demonstrated to activate both transcription factor nuclear factor kappaB (NFkappaB) and the c-Jun N-terminal kinase/stress activated protein kinase (JNK/SAPK) pathway, which in turn stimulates transcription factor activating protein 1 (AP1) mainly via phosphorylation of the c-Jun component. We have investigated the signaling properties of NFkappaB inducing kinase (NIK), a TRAF2-associated protein kinase that mediates NFkappaB induction. NIK was found to be unable to activate JNK/SAPK, mitogen-activated protein kinase, or p38 kinase. Moreover, NIK was not required for JNK/SAPK activation by TNF-R1, thus representing the first TNF-R1 complex component to dissect the NFkappaB and the JNK/SAPK pathways. Despite being unable to activate JNK/SAPK and mitogen-activated protein kinase, NIK strongly activated AP1 and was required for TNF-R1-induced AP1 activation. Therefore, NIK links TNF-R1 to a novel, JNK/SAPK-independent, AP1 activation pathway. PMID- 9334170 TI - Identification of a novel neural cell adhesion molecule-related gene with a potential role in selective axonal projection. AB - We describe here the cloning of mouse complementary DNAs encoding a novel protein, Rb-8 neural cell adhesion molecule (RNCAM), with a predicted extracellular region of five immunoglobulin C2-type domains followed by two fibronectin type III domains. Alternative splicing is likely to generate two RNCAM isoforms, which are differently attached to the cell membrane. These structural features and overall sequence identity identify this protein as a novel member of a cell adhesion molecule subgroup together with vertebrate neural cell adhesion molecule, Aplysia cell adhesion molecule, and Drosophila fasciclin II. In insects, fasciclin II is present on a restricted subset of embryonic central nervous system axons where it controls selective axon fasciculation. Intriguingly, RNCAM likewise is expressed in subsets of olfactory and vomeronasal neurons with topographically defined axonal projections. The spatial expression RNCAM corresponds precisely to that of certain odorant receptor expression zones of the olfactory epithelium. These expression patterns thus render RNCAM the first described cell adhesion molecule with a potential regulatory role in formation of selective axonal projections important for olfactory sensory information coding. PMID- 9334171 TI - The BST1 gene of Saccharomyces cerevisiae is the sphingosine-1-phosphate lyase. AB - Sphingolipids elicit a wide variety of eukaryotic cellular responses, most involving regulation of cell growth, differentiation, and apoptosis. Sphingosine 1-phosphate, a sphingolipid catabolite, is mitogenic in fibroblasts and inhibits the chemotactic mobility and invasiveness of human tumor cells. Sphingosine 1 phosphate degradation requires cleavage at the C2-3 carbon bond by sphingosine phosphate lyase. A yeast genetic approach was used to clone the first sphingosine phosphate lyase gene, BST1. BST1 overexpression conferred resistance to sphingosine in yeast. BST1 deletion produced sensitivity to exogenous D-erythro sphingosine and phytosphingosine and intracellular accumulation of sphingosine 1 phosphate upon exposure to exogenous sphingosine. This study confirms that sphingoid base metabolism is similar in all eukaryotes and suggests that yeast genetics may be useful in the isolation and identification of other genes involved in sphingolipid signaling and metabolism. PMID- 9334172 TI - Amyloid-specific heparan sulfate from human liver and spleen. AB - Heparan sulfate proteoglycans are consistently accumulated in tissues afflicted by amyloidosis and have been implicated in the mechanism of amyloid deposition. To study this relationship, heparan sulfate was isolated from liver and spleen of patients with AA amyloidosis and from normal organs and subjected to structural analysis. The polysaccharides were deaminated with nitrous acid, and the products were reduced with NaB3H4 to yield labeled oligosaccharides. Disaccharides obtained by selective deamination of intact or N-deacetylated polysaccharides were separated and quantified by anion-exchange high performance liquid chromatography and thus defined the composition of N-sulfated block regions or the entire heparan sulfate chains, respectively. The heparan sulfate samples derived from liver or spleen with AA-type amyloidosis were all similar in composition, regardless of tissue source, but differed from either control material. These findings suggest that secondary amyloidosis is associated with the deposition in the affected tissues of a heparan sulfate with a specifically modified structure. PMID- 9334173 TI - Thyroglobulin transport along the secretory pathway. Investigation of the role of molecular chaperone, GRP94, in protein export from the endoplasmic reticulum. AB - GRP94 serves as a molecular chaperone in the endoplasmic reticulum (ER). In normal thyrocytes, GRP94 interacts transiently with thyroglobulin (Tg), and in thyrocytes of animals suffering from congenital hypothyroid goiter with defective thyroglobulin, GRP94 and thyroglobulin associate in a protracted fashion. In order explore possible consequences of GRP94 binding, we have studied recombinant nonmutant thyroglobulin expressed in control Chinese hamster ovary (CHO) cells in comparison to that produced in CHO cells genetically manipulated for selectively increased GRP94 expression. Levels of ER chaperones other than GRP94 did not detectably differ, and thyroglobulin achieved transport competence in both kinds of CHO cells. However, increased availability of GRP94 caused the residence time of Tg in the ER to be remarkably prolonged. This was accompanied by a major increase in Tg directly associated with GRP94 and an increase in the ER pool size of Tg. Importantly, co-immunoprecipitation analysis revealed disulfide-linked Tg complexes (previously reported as an early Tg-folding intermediate) especially associated with GRP94. Indeed, non-native Tg, GRP94, and a 78-kDa protein likely to be BiP, appeared in ternary complexes. Under these conditions, GRP94 association appears directly involved in prolongation of Tg folding and export, consistent with a role in quality control in the ER. PMID- 9334174 TI - Active site-directed inhibitors of Rhodococcus 20 S proteasome. Kinetics and mechanism. AB - We have studied the mechanism of inhibition of the recombinant Rhodococcus proteasome by four different chemical classes of active site-directed small molecule inhibitors. Clasto-lactacystin beta-lactone is a time-dependent inhibitor of the Rhodococcus proteasome's ability to hydrolyze Suc-Leu-Leu-Val Tyr-AMC, a substrate for this proteasome's single type of active site, and proceeds with a kinact/[I] of 1,700 M-1 s-1. Using peptide mapping of tryptic digests, LC/MS, and amino acid sequence analysis, we have established that the Ogamma of the hydroxyl group on the N-terminal threonine of the beta-subunit is the sole modification made by the beta-lactone. Active site titrations of the Rhodococcus proteasome with reversible peptide aldehydes show the expected stoichiometry of one inhibitor molecule per beta-subunit. Prior modification with beta-lactone completely abrogates the binding of peptidyl boronic acid inhibitors, suggesting that these inhibitors also inactivate the enzyme by reacting with the Ogamma moiety on Thr1. High performance liquid chromatography analysis of peptidyl vinyl sulfone-modified intact Rhodococcus proteasome beta subunit and its tryptic peptides suggests that the peptidyl vinyl sulfone modifies a residue in the N-terminal 20 amino acids. This modification is also blocked by prior treatment with beta-lactone. PMID- 9334175 TI - An analysis of suppressor mutations suggests that the two halves of the lactose permease function in a symmetrical manner. AB - A conserved motif, GXXX(D/E)(R/K)XG[X](R/K)(R/K), is located in loop 2/3 and loop 8/9 in the lactose permease, and also in hundreds of evolutionarily related transporters. The importance of conserved residues in loop 8/9 was previously investigated (Pazdernik, N. J., Jessen-Marshall, A. E., and Brooker, R. J. (1997) J. Bacteriol. 179, 735-741). Although this loop was tolerant of many substitutions, a few mutations in the first position of the motif were shown to dramatically decrease lactose transport. In the current study, a mutant at the first position in the motif having very low lactose transport, Leu280, was used as a parental strain to isolate second-site revertants that restore function. A total of 23 independent mutants were sequenced and found to have a second amino acid substitution at several locations (G46C, G46S, F49L, A50T, L212Q, L216Q, S233P, C333G, F354C, G370C, G370S, and G370V). A kinetic analysis revealed that the first-site mutation, Leu280, had a slightly better affinity for lactose compared with the wild-type strain, but its Vmax for lactose transport was over 30-fold lower. The primary effect of the second-site mutations was to increase the Vmax for lactose transport, in some cases, to levels that were near the wild type value. When comparing this study to second-site mutations obtained from loop 2/3 defective strains, a striking observation was made. Mutations in three regions of the protein, codons 45-50, 234-241, and 366-370, were able to restore functionality to both loop 2/3 and loop 8/9 defects. These results are discussed within the context of a C1/C2 alternating conformation model in which lactose translocation occurs by a conformational change at the interface between the two halves of the protein. PMID- 9334176 TI - Probing the origin of acetyl-CoA and oxaloacetate entering the citric acid cycle from the 13C labeling of citrate released by perfused rat hearts. AB - We present a strategy for simultaneous assessment of the relative contributions of anaplerotic pyruvate carboxylation, pyruvate decarboxylation, and fatty acid oxidation to citrate formation in the perfused rat heart. This requires perfusing with a mix of 13C-substrates and determining the 13C labeling pattern of a single metabolite, citrate, by gas chromatography-mass spectrometry. The mass isotopomer distributions of the oxaloacetate and acetyl moieties of citrate allow calculation of the flux ratios: (pyruvate carboxylation)/(pyruvate decarboxylation), (pyruvate carboxylation)/(citrate synthesis), (pyruvate decarboxylation)/(citrate synthesis) (pyruvate carboxylation)/(fatty acid oxidation), and (pyruvate decarboxylation)/(fatty acid oxidation). Calculations, based on precursor-product relationship, are independent of pool size. The utility of our method was demonstrated for hearts perfused under normoxia with [U 13C3](lactate + pyruvate) and [1-13C]octanoate under steady-state conditions. Under these conditions, effluent and tissue citrate were similarly enriched in all 13C mass isotopomers. The use of effluent citrate instead of tissue citrate allows probing substrate fluxes through the various reactions non-invasively in the intact heart. The methodology should also be applicable to hearts perfused with other 13C-substrates, such as 1-13C-labeled long chain fatty acid, and under various conditions, provided that assumptions on which equations are developed are valid. PMID- 9334177 TI - A 13C mass isotopomer study of anaplerotic pyruvate carboxylation in perfused rat hearts. AB - Anaplerotic pyruvate carboxylation was examined in hearts perfused with physiological concentrations of glucose, [U-13C3]lactate, and [U-13C3]pyruvate. Also, a fatty acid, [1-13C]octanoate, or ketone bodies were added at concentrations providing acetyl-CoA at a rate resulting in either low or substantial pyruvate decarboxylation. Relative contributions of pyruvate and fatty acids to citrate synthesis were determined from the 13C labeling pattern of effluent citrate by gas chromatography-mass spectrometry (see companion article, Comte, B., Vincent, G., Bouchard, B., and Des Rosiers, C. (1997) J. Biol. Chem. 272, 26117-26124). Precision on flux measurements of anaplerotic pyruvate carboxylation depended on the mix of substrates supplied to the heart. Anaplerotic fluxes were precisely determined under conditions where acetyl-CoA was predominantly supplied by beta-oxidation, as it occurred with 0.2 or 1 mM octanoate. Then, anaplerotic pyruvate carboxylation provided 3-8% of the OAA moiety of citrate and was modulated by concentrations of lactate and pyruvate in the physiological range. Also, the contribution of pyruvate to citrate formation through carboxylation was equal to or greater than through decarboxylation. Furthermore, 13C labeling data on tissue citric acid cycle intermediates and pyruvate suggest that (i) anaplerosis occurs also at succinate and (ii) cataplerotic malate decarboxylation is low. Rather, the presence of citrate in the effluent perfusate of hearts perfused with physiological concentrations of glucose, lactate, and pyruvate and concentrations of octanoate leading to maximal oxidative rates suggests a cataplerotic citrate efflux from mitochondria to cytosol. Taken altogether, our data raise the possibility of a link between pyruvate carboxylation and mitochondrial citrate efflux. In view of the proposed feedback regulation of glycolysis by cytosolic citrate, such a link would support a role of anaplerosis and cataplerosis in metabolic signal transmission between mitochondria and cytosol in the normoxic heart. PMID- 9334178 TI - Identification, cloning, and mutational analysis of the casein kinase 1 cDNA of the malaria parasite, Plasmodium falciparum. Stage-specific expression of the gene. AB - The cDNA for casein kinase 1 (CK1) of Plasmodium falciparum was cloned, sequenced, and expressed in bacteria. The single major open reading frame of the 1.2-kilobase pair cDNA coded for a 324-amino acid polypeptide of approximately 37 kDa, the predicted sequence of which showed strong identity with known CK1 isoforms. The purified recombinant enzyme exhibited properties characteristic of CK1, such as inhibition by CK1-7, the ability to phosphorylate a highly specific peptide substrate, and a strong preference for ATP over GTP. A casein kinase activity, partially purified from soluble extracts of P. falciparum by affinity chromatography through CK1-7 columns displayed identical properties. The activity showed a stage-specific expression in the parasite, in the order trophozoite > ring >> schizont. Northern analysis indicated the existence of two major CK1 mRNAs, 2.4 and 3.2 kilobase pairs long, the levels of which were in the order ring > schizont > trophozoite. Mutagenesis of recombinant CK1 defined important amino acid residues and their potential role in the conformation of the enzyme. The malarial CK1 appeared to be the one of the smallest and perhaps the most primitive CK1 enzymes known, containing little sequence information beyond the minimal catalytic domain. PMID- 9334179 TI - The mechanism by which heparin promotes the inhibition of coagulation factor XIa by protease nexin-2. AB - Previous kinetic studies have shown that protease nexin-2 is a potent, reversible, and competitive inhibitor of factor XIa. Here we show that high molecular weight heparin potentiates the ability of protease nexin-2 to inhibit factor XIa with a parabolic concentration dependence, predominantly because of an increase of the association rate constant with little perturbation of the dissociation rate constant. No effect on factor XIa inhibition by protease nexin 2 was observed with heparin preparations of 6-22 saccharide units (0.1 nM-10 microM), whereas heparin preparations with 32-64 saccharide units potentiated factor XIa inhibition by protease nexin-2 in a size- and concentration-dependent manner. We propose a model wherein heparin exerts this effect by providing a template for the assembly of factor XIa-protease nexin-2 complexes, and only heparin polymers consisting of greater than 32 saccharide units (Mr approximately 10,000) are sufficiently long to provide a template to which factor XIa and protease nexin-2 molecules can bind simultaneously. Heparin-mediated enhancement of factor XIa inhibition by protease nexin-2 was partially abrogated by high molecular weight kininogen, suggesting that high molecular weight kininogen may play a role in regulating factor XIa activity. PMID- 9334180 TI - Intracellular sequestration of sodium by a novel Na+/H+ exchanger in yeast is enhanced by mutations in the plasma membrane H+-ATPase. Insights into mechanisms of sodium tolerance. AB - Sodium tolerance in yeast is disrupted by mutations in calcineurin, a Ca2+/calmodulin-dependent protein phosphatase, which is required for modulation of Na+ uptake and efflux mechanisms. Five Na+-tolerant mutants were isolated by selecting for suppressors of calcineurin mutations, and mapped to the PMA1 gene, encoding the plasma membrane H+-ATPase. One mutant, pma1-alpha4, which has the single amino acid change Glu367 --> Lys at a highly conserved site within the catalytic domain of the ATPase, was analyzed in detail to determine the mechanism of Na+ tolerance. After exposure to Na+ in the culture medium, 22Na influx in the pma1 mutant was reduced 2-fold relative to control, consistent with a similar decrease in ATPase activity. Efflux of 22Na from intact cells was relatively unchanged in the pma1 mutant. However, selective permeabilization of the plasma membrane revealed that mutant cells retained up to 80% of intracellular Na+ within a slowly exchanging pool. We show that NHX1, a novel gene homologous to the mammalian NHE family of Na+/H+ exchangers, is required for Na+ sequestration in yeast and contributes to the Na+-tolerant phenotype of pma1-alpha4. PMID- 9334181 TI - Interaction between Cdc42Hs and RhoGDI is mediated through the Rho insert region. AB - Members of the Rho subfamily of GTP-binding proteins contain a region of amino acid sequence (residues 122-134) that is absent from other Ras-like proteins and is termed the Rho insert region. To address the functional role of this domain, we have constructed a Cdc42Hs/Ras chimera in which loop 8 from Ha-Ras was substituted for the region in Cdc42Hs that contains the 13-amino acid insert region. Our data indicate that the insert region of Cdc42Hs is not essential for its interactions with various target/effector molecules or for interactions with the guanine nucleotide exchange factor, Dbl, or the Cdc42 GTPase-activating protein (GAP). However, the regulation of GDP dissociation and GTP hydrolysis on Cdc42Hs by the Rho GDP-dissociation inhibitor (GDI) is extremely sensitive to changes in the insert region, such that a Cdc42Hs/Ha-Ras chimera that lacks this insert is no longer susceptible to a GDI-induced inhibition of GDP dissociation and GTP hydrolysis. The insensitivity to GDI activity is not due to the inability of the GDI molecule to bind to the Cdc42Hs/Ha-Ras chimera, and in fact, the GDI is fully capable of stimulating the release of this chimera from membranes. PMID- 9334183 TI - Characterization of the Dictyostelium discoideum cellulose-binding protein CelB and regulation of gene expression. AB - Similar to other stages of Dictyostelium development, spore germination is a particularly suitable model for studying regulation of gene expression. The transition from spore to amoeba is accompanied by developmentally regulated changes in both protein and mRNA synthesis. A number of spore germination specific cDNAs have been isolated previously. Among these are two members of the 270 gene family, a group of four genes defined by the presence of a common tetrapeptide repeat of Thr-Glu-Thr-Pro. celA (formerly called 270-6) and celB (formerly 270-11) are expressed solely and coordinately during spore germination, the levels of the respective mRNAs being low in dormant spores, rising during germination to a maximum level at about 2 h, and then rapidly declining as amoebae are released from spores. The mRNAs are not found in growing cells or during multicellular development. The rapidity with which these transcripts accumulate and then disappear during germination implies that the respective products may be important for the process. We reported previously that the CelA protein is a cellulase (endo-1, 4-beta-glucanase (EC 3.2.1.4)). In the present investigation, properties of the CelB protein, a glycosylated protein of 532 amino acids, 36% of which are serine or threonine, were examined, and the upstream sequences involved in the developmental regulation of the expression of the gene have been determined. The CelB protein does not demonstrate cellulase activity, but it has a cellulose-binding domain. Its role, if any, in degradation of the cellulose-containing spore wall is unknown. To identify cis-acting elements in the celB promoter, unidirectional 5' deletions of the celB upstream noncoding region were constructed and used to transform amoebae. Analysis of promoter activity during different stages of development shows that a short, very A/T-rich sequence of approximately 81 base pairs is sufficient for spore-specific celB transcription. Contained in this sequence is the Myb oncogene protein binding site, TAACTG, which was shown previously to be a negative regulator of celA transcription. Dictyostelium and mouse Myb proteins bind to this region of the promoter, suggesting that Myb might regulate celB gene expression negatively as it does in celA. PMID- 9334182 TI - Appearance of phosphatidylserine on apoptotic cells requires calcium-mediated nonspecific flip-flop and is enhanced by loss of the aminophospholipid translocase. AB - Phosphatidylserine (PS), ordinarily sequestered in the plasma membrane inner leaflet, appears in the outer leaflet during apoptosis, where it triggers non inflammatory phagocytic recognition of the apoptotic cell. The mechanism of PS appearance during apoptosis is not well understood but has been associated with loss of aminophospholipid translocase activity and nonspecific flip-flop of phospholipids of various classes. The human leukemic cell line HL-60, the T cell line Jurkat, and peripheral blood neutrophils, undergoing apoptosis induced either with UV irradiation or anti-Fas antibody, were probed in the cytofluorograph for (i) surface PS using fluorescein isothiocyanate-labeled annexin V, (ii) PS uptake by the aminophospholipid translocase using [6-[(7 nitrobenz-2-oxa-1,3-diazol-4-yl)amino] caproyl] (NBD)-labeled PS, (iii) nonspecific uptake of phospholipids (as a measure of transbilayer flip-flop) using NBD-labeled phosphatidylcholine, and (iv) the appearance of hypodiploid DNA. In all three types of cells undergoing apoptosis, the appearance of PS followed loss of aminophospholipid translocase and was accompanied by nonspecific phospholipid flip-flop. Importantly, however, in the absence of extracellular calcium, the appearance of PS was completely inhibited despite DNA fragmentation and loss of aminophospholipid translocase activity, the latter demonstrating that loss of the translocase is insufficient for PS appearance during apoptosis. Furthermore, while both the appearance of PS and nonspecific phospholipid uptake demonstrated identical extracellular calcium requirements with an ED50 of nearly 100 microM, the magnitude of PS appearance depended on the level of aminophospholipid translocase activity. Taken together, the data strongly suggest that while nonspecific flip-flop is the driving event for PS appearance in the plasma membrane outer leaflet, aminophospholipid translocase activity ultimately modulates its appearance. PMID- 9334184 TI - Erythropoietin induces the tyrosine phosphorylation of insulin receptor substrate 2. An alternate pathway for erythropoietin-induced phosphatidylinositol 3-kinase activation. AB - In this report, we demonstrate that insulin receptor substrate-2 (IRS-2) is phosphorylated on tyrosine following treatment of UT-7 cells with erythropoietin. We have investigated the expression of IRS-1 and IRS-2 in several cell lines with erythroid and/or megakaryocytic features, and we observed that IRS-2 was expressed in all cell lines tested. In contrast, we did not detect the expression of IRS-1 in these cells. In response to erythropoietin, IRS-2 was immediately phosphorylated on tyrosine, with maximal phosphorylation between 1 and 5 min. Tyrosine-phosphorylated IRS-2 was associated with phosphatidylinositol 3-kinase and with a 140-kDa protein that comigrated with the phosphatidylinositol-3,4,5 trisphosphate 5-phosphatase, SHIP. Moreover, IRS-2 was constitutively associated with the erythropoietin receptor. We did not observe the association of IRS-2 with JAK2, Grb2, or PTP1D. Using BaF3 cells transfected with mutated erythropoietin receptors, we demonstrate that neither the tyrosine residues of the intracellular domain nor the last 109 amino acids of the erythropoietin receptor are required for erythropoietin-induced IRS-2 tyrosine phosphorylation. Altogether, our results indicate that erythropoietin-induced IRS-2 tyrosine phosphorylation could account for the previously reported activation of phosphatidylinositol 3-kinase mediated by erythropoietin receptors mutated in the phosphatidylinositol 3-kinase-binding site (Damen, J., Cutler, R. L., Jiao, H., Yi, T., and Krystal, G. (1995) J. Biol. Chem. 270, 23402-23406; Gobert, S., Porteu, F., Pallu, S., Muller, O., Sabbah, M., Dusanter-Fourt, I., Courtois, G., Lacombe, C., Gisselbrecht, S., and Mayeux, P. (1995) Blood 86, 598-606). PMID- 9334185 TI - Domain structures and immunogenic regions of the 90-kDa heat-shock protein (HSP90). Probing with a library of anti-HSP90 monoclonal antibodies and limited proteolysis. AB - Domain structures of the 90-kDa heat-shock protein (HSP90) have been investigated with a library of anti-HSP90 monoclonal antibodies (mAbs) and by limited proteolysis with trypsin and chymotrypsin. Thirty-three mAbs were obtained by immunization with bacterially expressed human HSP90alpha and HSP90beta isoforms. Among them, ten and three mAbs reacted specifically with HSP90alpha and HSP90beta, respectively. Immunoblotting and enzyme-linked immunosorbent analyses revealed that major immunogenic domains were located at two restricted regions of HSP90alpha, i.e. amino acids 227-310 (designated Region I) and 702-716 (Region II), corresponding to a highly charged region and a region near the C terminus, respectively. Taken together with the characteristics of the amino acid sequences, these two immunogenic regions appeared to be exposed at the outer surface of HSP90. We further investigated the domain structures of HSP90 by limited proteolysis in combination with N-terminal sequencing and immunoblotting analyses. Tryptic cleavages of HSP90alpha at low concentrations revealed the existence of major susceptible sites at Arg400-Glu401, Lys615-Ala616, and Arg620 Asp621. Proteolysis at higher trypsin concentrations caused successive cleavages only toward the N-terminal direction from these sites, and Region I was included in the region selectively deleted during this process, thereby further suggesting its surface location. From these results, we propose three domain structures of HSP90 consisting of amino acids 1-400, 401-615, and 621-732. Differences in the protease sensitivity and immunogenicity further suggest that every domain is composed of two subdomains. This is the first study describing the domain structures and the immunogenic regions of HSP90. PMID- 9334186 TI - The Ets transcription factors interact with each other and with the c-Fos/c-Jun complex via distinct protein domains in a DNA-dependent and -independent manner. AB - The transcription factors Fos, Jun, and Ets regulate the expression of human stromelysin-1 and collagenase-1 genes. Recently, we found that ERG, an Ets family member, activates collagenase-1 gene but not stromelysin-1 by physically interacting with c-Fos/c-Jun. Interestingly, ERG binds to stromelysin-1 promoter and represses its activation by ETS2. Here, to investigate the molecular mechanism of this regulation, we have used an in vitro protein-protein interaction assay and studied the transcription factor interactions of ETS2. We found that ETS2 could weakly associate with in vitro synthesized ETS1, c-Fos, and c-Jun and strongly with c-Fos/c-Jun complex and ERG via several distinct ETS2 domains including the C-terminal region that contains the DNA-binding domain. Strikingly, these interactions were stabilized in vitro by DNA as they were inhibited by ethidium bromide. Both the N-terminal region, comprising the transactivation domain, and the C-terminal region of ETS2 associated with ERG and, interestingly, the interaction of ERG through the transactivation domain of ETS2 was DNA-independent. The DNA-dependent interaction of ETS2 with c-Fos/c-Jun was enhanced by specific DNA fragments requiring two Ets-binding sites of the stromelysin-1 promoter. Using the two hybrid system, we also demonstrated that ETS2 interacts with c-Jun or ERG in vivo. PMID- 9334187 TI - Differential affinity cross-linking of phosphorylase kinase conformers by the geometric isomers of phenylenedimaleimide. AB - Phosphorylase b kinase (PbK) from skeletal muscle is a highly regulated oligomer consisting of four copies of four distinct subunits (alphabetagamma)delta4. The gamma subunit is catalytic, and the remaining subunits are regulatory. To characterize effector-induced changes in the quaternary structure of the enzyme, we utilized the ortho-, meta, and para-isomers of phenylenedimaleimide (PDM), which in addition to having different geometries, also vary 2.5-fold in their cross-linking spans. Even at concentrations equivalent to the alphabetagammadelta protomers of PbK, all three isomers caused specific, rapid, and extensive cross linking of the holoenzyme to form primarily alphabeta dimers, plus smaller amounts of betagammagamma and alphagammagamma trimers. The formation of these three conjugates was nearly totally inhibited by a 10-fold molar excess over PDM of N-(o- and p-tolyl)succinimide, which are chemically inert structural analogs of PDM. This inhibition suggests that PbK has binding sites for PDM and that PDM acts as an affinity cross-linker in binding to these sites prior to forming cross linked conjugates. The largest effect on cross-linking in progressing from o- to p-PDM was on the alphagammagamma trimer, which is preferentially formed by the p isomer. Activation of the enzyme by either phosphorylation or the allosteric activators ADP and GDP resulted in large increases in the amount of alphagammagamma formed, small increases in betagammagamma, and little change in alphabeta. When cross-linked in the presence of the reversibly activating nucleoside diphosphates, PbK remained activated after their removal, indicating that cross-linking had locked it in the active conformation. Our results provide direct evidence for perturbations in the interactions of the catalytic gamma subunit with the regulatory alpha and beta subunits upon activation of PbK. PMID- 9334189 TI - Alpha1-antitrypsin Portland inhibits processing of precursors mediated by proprotein convertases primarily within the constitutive secretory pathway. AB - We studied the extent of cellular inhibitory activity of alpha1-antitrypsin Portland (alpha1-PDX), a potent inhibitor of proprotein convertases of the subtilisin/kexin type. We compared the inhibitory effects of alpha1-PDX on the intracellular processing of two model precursors (pro-7B2 and POMC) mediated by six of the seven known mammalian convertases, namely furin, PC1, PC2, PACE4, PC5 A, PC5-B, and PC7. The substrates selected were pro7B2, a precursor cleaved within the trans-Golgi network (TGN), and pro-opiomelanocortin, which is processed in the TGN and secretory granules. Biosynthetic analyses were performed using either vaccinia virus expression in BSC40, GH4C1, and AtT20 cells, or stable transfectants of alpha1-PDX in AtT20 cells. Results revealed that alpha1 PDX inhibits processing of these precursors primarily within the constitutive secretory pathway and that alpha1-PDX is cleaved into a shorter form by some convertases. Evidence is presented demonstrating that in contrast to the full length alpha1-PDX (64 kDa), the cleaved (56 kDa) secreted product does not significantly inhibit furin activity in vitro. Cellular expression of alpha1-PDX results in modified contents of mature secretory granules with increased levels of partially processed products. Biosynthetic and immunocytochemical analyses of AtT20/alpha1-PDX cells demonstrated that alpha1-PDX is primarily localized within the TGN, and that a small proportion enters secretory granules where it is mostly stored as the cleaved product. PMID- 9334188 TI - The structural effects of endogenous and exogenous Ca2+/calmodulin on phosphorylase kinase. AB - The activity of phosphorylase b kinase (PbK) is stimulated by Ca2+ ions, which act through its endogenous calmodulin subunit (delta), and further stimulated by the Ca2+-dependent binding of exogenous calmodulin (delta'). In contrast to their highly characterized effects on activity, little is known regarding the structural effects on the (alphabetagammadelta)4 PbK holoenzyme induced by Ca2+ and delta'/Ca2+. We have used mono- and bifunctional chemical modifiers as conformational probes to compare how the two effectors influence the structure of the catalytic gamma subunit and the interactions among all of the subunits. As determined by reductive methylation and carboxymethylation, Ca2+ increased the accessibility of the gamma subunit; it also increased the formation by phenylenedimaleimide of an alphagammagamma conjugate that is characteristic of activated conformations of PbK (Nadeau, O. W., Sacks, D. M., and Carlson, G. M. (1997) J. Biol. Chem. 272, 26196-26201); however, Ca2+ also had structural effects that were clearly distinct from other activators. Moreover, similar structural effects of Ca2+ were observed with PbK that had been activated by phosphorylation, consistent with the fact that such activation does not eliminate the catalytic dependence of the enzyme on Ca2+. Our results suggest tiers of conformational transitions in the activation of PbK, with the most fundamental being induced by Ca2+. Analysis of the various cross-linked conjugates formed in the presence of Ca2+ by o-phenylenedimaleimide or m-maleimidobenzoyl-N hydroxysuccinimide ester showed that the binding of Ca2+ to the delta subunit triggers changes in the interactions among all subunits, including between protomers, indicating an extensive communication network throughout the PbK complex. Most of the structural effects of delta'/Ca2+ were qualitatively similar to, but quantitatively greater than, the effects of Ca2+ alone; but delta'/Ca2+ also had distinct effects, especially involving cross-linking of the delta subunit. PMID- 9334190 TI - Interaction of phosphorylated tryptophan hydroxylase with 14-3-3 proteins. AB - Rabbit brain tryptophan hydroxylase (TPH) has been expressed in insect cells (Spodoptera frugiperda) as a histidine-tagged enzyme. The specific activity of the purified fusion enzyme is 80 nmol of 5-hydroxytryptophan/min/mg. Multifunctional regulatory 14-3-3 proteins were purified from fresh bovine brain. Phosphorylation and 14-3-3 proteins play important roles in the regulation of TPH activity. We have found that phosphorylation of TPH by cAMP-dependent protein kinase increased the activity of the hydroxylase by 25-30% and that 14-3-3 proteins increased the hydroxylase activity of phosphorylated TPH by approximately 45%. Under these conditions, the 14-3-3 proteins were not phosphorylated, and unphosphorylated TPH was not activated by 14-3-3 proteins. Surface plasmon resonance analysis demonstrated that 14-3-3 proteins bind to phosphorylated TPH with an affinity constant (Ka) of 4.5 x 10(7) M-1. Binding studies using affinity chromatography also showed that 14-3-3 proteins interact with phosphorylated TPH. The dephosphorylation of TPH by protein phosphatase-1 was inhibited by 14-3-3 proteins. Our results demonstrate that 14-3-3 proteins form a complex with phosphorylated brain TPH, thereby increasing its enzymatic activity and inhibiting its dephosphorylation. PMID- 9334191 TI - Evidence of the indirect formation of the catecholic intermediate substrate responsible for the autoactivation kinetics of tyrosinase. AB - Tyrosinase (EC 1.14.18.1) exhibits unusual kinetic properties in the oxidation of monohydric phenol substrates consisting of a lag period that increases with increasing substrate concentration. The cause of this is an autocatalytic process dependent on the generation of a dihydric phenol substrate, which acts as an activator of the enzyme. Experiments with N-substituted dihydric phenol substrates (N-methyldopamine, N-acetyldopamine) demonstrate that oxygen consumption is retarded in the N-acetyl substituted material due to a diminished rate of cyclization. The oxygen uptake exhibited a similar pattern when N acetyltyramine was oxidized, and this was reflected by a prolongation of the lag period. N,N-Dipropyldopamine was oxidized with normal kinetics but with an oxygen stoichiometry of 0.5 mol of oxygen/mol of substrate. We show that this is the result of the formation of a stable indoliumolate product with oxidation reduction properties that prevent the formation of dopaminochrome, thus blocking further stages in the tyrosinase-catalyzed oxidation. Evidence that the indoliumolate product is formed by cyclization of the ortho-quinone is presented by pulse radiolysis studies, which demonstrate the formation of the ortho-quinone (by disproportionation of the corresponding semiquinones), which cyclizes to give the indoliumolate. The rate constant for cyclization was shown to be 48 s-1 (at pH 6.0). Tyrosinase-catalyzed oxidation of the monohydric phenol analogue, N, N dimethyltyramine, was shown to require the addition of a dihydric phenol. Oxygen utilization then exhibited a stoichiometry of 1.0, indicating that the reactions proceed only as far as the cyclization. The analogous stable cyclic indoliumolate product was shown to be formed, with UV absorption and NMR spectra closely similar to the indoliumolate derived from N,N-dipropyldopamine. This material was methylated by catechol O-methyltransferase but was unreactive to redox reagents. The formation of the cyclic product accounts for the indefinite lag when N,N dimethyltyramine is used as the substrate for tyrosinase in the absence of a dihydric phenol cofactor. PMID- 9334192 TI - Activation of the megakaryocyte-specific gene platelet basic protein (PBP) by the Ets family factor PU.1. AB - Platelet basic protein (PBP) is a chemokine family member that is only found in platelets and their precursors megakaryocytes. The PBP gene is physically linked to the gene for another platelet-specific chemokine, platelet factor 4. While the biological basis of platelet factor 4 expression has been pursued by others, the regulatory features controlling the platelet-specific expression of PBP have not been investigated. In this article, we examined the molecular basis by which this megakaryocyte-specific gene is regulated. Transient expression studies of truncated reporter constructs containing from 4.5 to 0.1 kilobases of the functional PBP gene 5'-flanking region, demonstrated that the proximal 0.1 kilobases of the promoter was sufficient for high levels of expression in human erythroleukemia and CHRF-288 cells, two megakaryocytic cell lines. However, none of these constructs was expressed above background levels in HeLa and 293 cells, two non-megakaryocytic cell lines. Further truncation of this promoter suggested that there was an important regulatory element(s) within a pyrimidine-rich tract. Mobility shift analysis of the pyrimidine-rich tract defined a region between -85 and -64 which bound to a nuclear factor(s). This region contains sequences matching the consensus Ets-binding site from -78 to -75 base pairs. In particular, we noted that this site matched a PU.1 consensus sequence known as a PU box. Mobility shift and supershift studies with nuclear extracts as well as recombinant PU.1 protein and anti-PU.1 antibody further confirmed that PU.1 was the specific Ets family factor that bound to this site. Transient expression assays using reporter constructs which contained point mutations that abrogated PU.1 binding also significantly reduced PBP promoter activity in human erythroleukemia and CHRF cells. In addition, while all reporter gene constructs containing PBP promoters were completely inactive in HeLa cells, transactivation experiments using a PU.1 expression construct demonstrated that exogenous expression of PU.1 could increase reporter gene expression up to 8-fold in these cells. Finally, the role of PU.1 in PBP gene expression was compared between wild type and PU.1-null embryonic stem (ES) cells that were differentiated in vitro into cells that resembled megakaryocytes both morphologically and immunologically. We found that PBP gene expression in the differentiated PU.1(-/ ) null ES cells (as determined by semi-quantitative reverse transcriptase polymerase chain reaction) was more than four times lower than that in the wild type ES cells, while other platelet-specific genes were expressed equally or similarly in the two ES cell lines. Previous reports have shown that PU.1 is expressed in several hematopoietic lineages, including megakaryocytes. However, the functional role of PU.1 has only been previously demonstrated in the myeloid and lymphoid lineages. Therefore, our studies are the first to show the biological importance of this nuclear factor in the regulated expression of a megakaryocyte-specific gene. PMID- 9334193 TI - Monochloramine inhibits phorbol ester-inducible neutrophil respiratory burst activation and T cell interleukin-2 receptor expression by inhibiting inducible protein kinase C activity. AB - Monochloramine derivatives are long lived physiological oxidants produced by neutrophils during the respiratory burst. The effects of chemically prepared monochloramine (NH2Cl) on protein kinase C (PKC) and PKC-mediated cellular responses were studied in elicited rat peritoneal neutrophils and human Jurkat T cells. Neutrophils pretreated with NH2Cl (30-50 microM) showed a marked decrease in the respiratory burst activity induced by phorbol 12-myristate 13-acetate (PMA), which is a potent PKC activator. These cells, however, were viable and showed a complete respiratory burst upon arachidonic acid stimulation, which induces the respiratory burst by a PKC-independent mechanism. The NH2Cl-treated neutrophils showed a decrease in both PKC activity and PMA-induced phosphorylation of a 47-kDa protein, which corresponds to the cytosolic factor of NADPH oxidase, p47(phox). Jurkat T cells pretreated with NH2Cl (20-70 microM) showed a decrease in the expression of the interleukin-2 receptor alpha chain following PMA stimulation. This was also accompanied by a decrease in both PKC activity and nuclear transcription factor-kappaB activation, also without loss of cell viability. These results show that NH2Cl inhibits PKC-mediated cellular responses through inhibition of the inducible PKC activity. PMID- 9334194 TI - Elimination of cholesterol in macrophages and endothelial cells by the sterol 27 hydroxylase mechanism. Comparison with high density lipoprotein-mediated reverse cholesterol transport. AB - Cultured macrophages and endothelial cells have been reported to secrete 27 oxygenated metabolites of cholesterol. This mechanism was compared with the classical high density lipoprotein (HDL)-dependent reverse cholesterol transport. Under standard conditions, macrophage preparations had considerably higher capacity to secrete 27-hydroxycholesterol and 3beta-hydroxy-5-cholestenoic acid than had endothelial cells and fibroblasts. Western blotting showed that lung macrophages contained the most sterol 27-hydroxylase protein of the cells tested. The relative amounts of 3beta-hydroxy-5-cholestenoic acid produced by the macrophages were also highest. Macrophages derived from monocytes of patients with sterol 27-hydroxylase deficiency did not secrete 27-oxygenated products, demonstrating that sterol 27-hydroxylase is the critical enzyme for the conversion of cholesterol into the 27-oxygenated steroids. That sterol 27 hydroxylase is responsible not only for 27-hydroxylation of cholesterol but also for the further oxidation of this steroid into 3beta-hydroxy-5-cholestenoic acid was shown with use of tritium-labeled 27-hydroxycholesterol and an inhibitor of sterol 27-hydroxylase. Secretion of 27-oxygenated products by the cultured macrophages as well as the ratio between the alcohol and the acid appeared to be dependent upon total 27-hydroxylase activity, the availability of substrate cholesterol, and the presence of an acceptor for 27-hydroxycholesterol in the medium. With albumin as extracellular acceptor, the major secreted product was 3beta-hydroxy-5-cholestenoic acid. Under such conditions, secretion of labeled 27 oxygenated products was higher than that of labeled cholesterol from lung alveolar macrophages preloaded with [4-14C]cholesterol. With HDL as acceptor, 27 hydroxycholesterol was the major secreted product, and the total secretion of labeled 27-oxygenated products was only about 10% of that of labeled cholesterol. Thus, 27-hydroxycholesterol and cholesterol may compete for HDL-mediated efflux from the cells. The results support the contention that the sterol 27-hydroxylase mediated elimination of cholesterol is more important in macrophages than in endothelial cells. This mechanism may be an alternative and/or a complement to the classical HDL-mediated reverse cholesterol transport in macrophages, in particular when the concentration of HDL is low. PMID- 9334195 TI - Primary structure of a new phosphocholine-containing glycoglycerolipid of Mycoplasma fermentans. AB - The chemical structure of a novel phosphocholine-containing glycoglycerolipid, the major polar lipid in the cell membrane of Mycoplasma fermentans PG18, was investigated by chemical analyses, gas-liquid chromatography-mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, as well as one- and two-dimensional homo- and heteronuclear NMR spectroscopy and identified as 6'-O-(3"-phosphocholine-2"-amino-1"-phospho-1", 3" propanediol)-alpha-D-glucopyranosyl-(1'-->3)-1,2-diacyl-glycerol (MfGL-II). Palmitate (16:0) and stearate (18:0), in a 3.6:1 molar ratio, constitute the major fatty acids present. MALDI-TOF mass spectrometry revealed two major pseudomolecular ions at m/z 1049.5 [MI + H]+ and 1077.3 [MII + H]+ representing a dipalmitoyl as the major component and a palmitoyl-stearoyl structure as a minor component. This is the first report of 2-amino-1,3-propanediol-1,3-bisphosphate present in a natural product. This glycoglycerolipid is the second phosphocholine containing glycoglycerolipid found in M. fermentans. PMID- 9334196 TI - Modification of alpha-chain or beta-chain heme pocket polarity by Val(E11) --> thr substitution has different effects on the steric, dynamic, and functional properties of human recombinant hemoglobin. Deoxy derivatives. AB - The dynamic and functional properties of mutant deoxyhemoglobins in which either the beta-globin Val67(E11) or the alpha-globin Val62(E11) is replaced by threonine have been investigated through the thermal evolution of the Soret absorption band in the temperature range 300 to 20 K and through the kinetics of CO rebinding after flash photolysis at room temperature. The conformational properties of the modified alpha chain and beta chain distal heme pockets were also studied through x-ray crystallography and molecular modeling. The data obtained with the various techniques consistently indicate that the polar isosteric mutation in the distal side of the alpha chain heme pocket has a larger effect on the investigated properties than the analogous mutation on the beta chain. We attribute the observed differences to the presence of a water molecule in the distal heme pocket of the modified alpha chains, interacting with the hydroxyl of the threonine side chain. This is indicated by molecular modeling which showed that the water molecule present in the alpha chain distal heme pocket can bridge by H bonding between Thr62(E11) and His58(E7) without introducing any unfavorable steric interactions. Consistent with the dynamic and functional data, the presence of a water molecule in the distal heme pocket of the modified beta chains is not observed by x-ray crystallography. PMID- 9334197 TI - The interaction between the endothelial cell protein C receptor and protein C is dictated by the gamma-carboxyglutamic acid domain of protein C. AB - The endothelial cell protein C receptor (EPCR) binds to both protein C and activated protein C (APC) with similar affinity. Removal of the Gla domain of protein C results in the loss of most of the binding affinity. This observation is compatible with at least two models: 1) the Gla domain of protein C interacts with phospholipid on cell surfaces to stabilize interaction with EPCR or 2) the Gla domain of protein C makes specific protein-protein interactions with EPCR. The latter model predicts that chimeric proteins containing the protein C Gla domain should interact with EPCR. To test this, we constructed a prothrombin chimera in which the Gla domain and aromatic stack of prothrombin were replaced with the corresponding region of protein C. The 125I-labeled chimera (Kd = 176 nM) and 125I-APC (Kd = 65 nM) both bound specifically to 293 cells stably transfected with EPCR, but both bound poorly to sham-transfected cells. The chimera also blocked APC binding to EPCR-transfected cells in a dose-dependent fashion (Ki approximately 139 nM) similarly to protein C (Ki approximately 75 nM). Chimera binding to EPCR-transfected cells was blocked by soluble EPCR, demonstrating direct protein-protein interaction between the chimera and EPCR. Consistent with this conclusion, the isolated Gla domain of protein C blocked APC binding to EPCR-transfected cells (IC50 = 2 microM). No inhibition was observed with the isolated prothrombin Gla domain. A protein C chimera with the prothrombin Gla domain and aromatic stack failed to bind to EPCR detectably. These data suggest that the Gla domain of protein C is responsible for much of the binding energy and specificity of the protein C-EPCR interaction. PMID- 9334199 TI - Major changes in the kinetic mechanism of AMP inhibition and AMP cooperativity attend the mutation of Arg49 in fructose-1,6-bisphosphatase. AB - The significance of subunit interface residues Arg49 and Lys50 in the function of porcine liver fructose-1,6-bisphosphatase was explored by site-directed mutagenesis, initial rate kinetics, and circular dichroism spectroscopy. The Lys50 --> Met mutant had kinetic properties similar to the wild-type enzyme but was more thermostable. Mutants Arg49 --> Leu, Arg49 --> Asp, Arg49 --> Cys were less thermostable than the wild-type enzyme yet exhibited wild-type values for kcat and Km. The Ki for the competitive inhibitor fructose 2,6-bisphosphate increased 3- and 5-fold in Arg49 --> Leu and Arg49 --> Asp, respectively. The Ka for Mg2+ increased 4-8-fold for the Arg49 mutants, with no alteration in the cooperativity of Mg2+ binding. Position 49 mutants had 4-10-fold lower AMP affinity. Most significantly, the mechanism of AMP inhibition with respect to fructose 1,6-bisphosphate changed from noncompetitive (wild-type enzyme) to competitive (Arg49 --> Leu and Arg49 --> Asp mutants) and to uncompetitive (Arg49 --> Cys mutant). In addition, AMP cooperativity was absent in the Arg49 mutants. The R and T-state circular dichroism spectra of the position 49 mutants were identical and superimposable on only the R-state spectrum of the wild-type enzyme. Changes from noncompetitive to competitive inhibition by AMP can be accommodated within the framework of a steady-state Random Bi Bi mechanism. The appearance of uncompetitive inhibition, however, suggests that a more complex mechanism may be necessary to account for the kinetic properties of the enzyme. PMID- 9334200 TI - Tyrosine-dependent basolateral sorting signals are distinct from tyrosine dependent internalization signals. AB - Converting cysteine 543 to tyrosine in the influenza virus hemagglutinin (HA) introduces both a basolateral sorting signal and an internalization signal into the HA cytoplasmic domain. Another HA mutant, HA+8, contains eight additional amino acids at the end of the cytoplasmic domain that include a powerful internalization signal. HA+8 was also sorted efficiently to the basolateral surface of Madin-Darby canine kidney cells. The simplest explanation for the observation that multiple sorting phenotypes depend upon the same small amino acid sequence is that certain tyrosine-based internalization signals might also function as basolateral sorting signals. To test this hypothesis, second-site mutations were introduced into HA C543Y or HA+8 to determine if the internalization and basolateral sorting functions can be separated. For HA C543Y, the same sequence positions were important for both basolateral sorting and internalization, but the two functions responded differently to individual amino acid replacements, indicating that they were distinct. For HA+8, the basolateral sorting signal required the same tyrosine as the internalization signal, but did not share any other characteristics. Thus, even when basolateral sorting signals that depend on tyrosine overlap or are co-linear with internalizations signals, the two sorting processes are sensitive to different characteristics of the sequence. PMID- 9334198 TI - TGT3, thyroid transcription factor I, and Sp1 elements regulate transcriptional activity of the 1.3-kilobase pair promoter of T1alpha, a lung alveolar type I cell gene. AB - Alveolar type I epithelial cells form the major surface for gas exchange in the lung. To explore how type I cells differ in gene expression from their progenitor alveolar type II cells, we analyzed transcriptional regulation of T1alpha, a gene expressed by adult type I but not type II cells. In vivo developmental patterns of T1alpha expression in lung and brain suggest active gene regulation. We cloned and sequenced 1.25 kilobase pairs of the T1alpha promoter that can drive reporter expression in lung epithelial cell lines. Deletion analyses identified regions important for lung cell expression. The base pair (bp) -100 to -170 fragment conferred differential regulation in lung epithelial cells compared with fibroblasts. Sequence alignment of this fragment with type II-specific surfactant protein B and C promoters shows similar consensus elements arranged in a different order. Gel retardation studies with alveolar epithelial cell line nuclear extracts, thyroid transcription factor I (TTF-1) homeodomain, hepatic nuclear factor (HNF)-3beta, or Sp1 proteins, and supershift assays were used to characterize TTF-1, HNF-3 (TGT3), and Sp1/Sp3 binding sites. The TGT3 site binds factors with binding properties similar to HNF-3/Fkh (hepatic nuclear factor 3/forkhead) proteins but different from HNF-3alpha or HNF-3beta. Co-transfection with a TTF-1 expression vector moderately transactivated the -170 bp-reporter construct. Mutational analysis of these three binding sites showed reduced transcriptional activity of the -170 bp promoter. Therefore, several regulatory sequences involved in type II cell gene regulation are also present in the T1alpha promoter, suggesting that genes of the peripheral lung epithelium may be regulated by similar factors. PMID- 9334201 TI - Role of the isoforms of CCAAT/enhancer-binding protein in the initiation of phosphoenolpyruvate carboxykinase (GTP) gene transcription at birth. AB - The gene for phosphoenolpyruvate carboxykinase (PEPCK), a target of CCAAT/enhancer-binding protein-alpha (C/EBPalpha) and -beta (C/EBPbeta), begins to be expressed in the liver at birth. Mice homozygous for a deletion in the gene for CEBPalpha (C/EBPalpha-/- mice) die shortly after birth of hypoglycemia, with no detectable hepatic PEPCK mRNA and negligible hepatic glycogen stores. Half of the mice homozygous for a deletion in the gene for CEBPbeta (C/EBPbeta-/- mice) have normal glucose homeostasis (phenotype A), and the other half die at birth of hypoglycemia due to a failure to express the gene for PEPCK and to mobilize hepatic glycogen (phenotype B). Insulin deficiency induces C/EBPalpha and PEPCK gene transcription in the livers of 19-day fetal rats, whereas dibutyryl cyclic AMP (Bt2cAMP) increases the expression of the gene for C/EBPbeta and causes a transient burst of PEPCK mRNA. Bt2cAMP induces PEPCK mRNA in the livers of fetal C/EBPalpha-/- mice, but at only 20% of the level of control animals; however, there is no induction of PEPCK mRNA if the cyclic nucleotide is injected into C/EBPalpha-/- mice immediately after delivery. The expression of the gene for C/EBPbeta is markedly induced in the livers of C/EBPalpha-/- mice within 2 h after the administration of Bt2cAMP. C/EBPbeta-/- mice injected at 20 days of fetal life with Bt2cAMP have a normal pattern of induction of hepatic PEPCK mRNA. In C/EBPbeta-/- mice with phenotype B, the administration of Bt2cAMP immediately after delivery induces PEPCK mRNA, causes the mobilization of hepatic glycogen, and maintains normal glucose homeostasis for up to 4 h (duration of the experiment). We conclude that C/EBPalpha is required for the cAMP induction of PEPCK gene expression in the liver and that C/EBPbeta can compensate for the loss of C/EBPalpha if its concentration is induced to appropriate levels. PMID- 9334202 TI - A role for nuclear phosphatidylinositol-specific phospholipase C in the G2/M phase transition. AB - Protein kinase C (PKC) is activated at the nucleus during the G2 phase of cell cycle, where it is required for mitosis. However, the mechanisms controlling cell cycle-dependent activation of nuclear PKC are not known. We now report that nuclear levels of the major physiologic PKC activator diacylglycerol (DAG) fluctuate during cell cycle. Specifically, nuclear DAG levels in G2/M phase cells are 2. 5-3-fold higher than in G1 phase cells. In synchronized cells, nuclear DAG levels rise to a peak coincident with the G2/M phase transition and return to basal levels in G1 phase cells. This increase in DAG level is sufficient to stimulate betaII PKC-mediated phosphorylation of its mitotic nuclear envelope substrate lamin B in vitro. Isolated nuclei from G2 phase cells contain an active phospholipase activity capable of generating DAG in vitro. Nuclear phospholipase activity is inhibited by the selective phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor 1-O-octadeyl-2-O-methyl-sn-glycero-3 phosphocholine and neomycin sulfate, but not by the phosphatidylcholine-PLC selective inhibitor D609 or inhibitors of phospholipase D-mediated DAG generation. Treatment of synchronized cells with 1-O-octadeyl-2-O-methyl-sn glycero-3-phosphocholine leads to decreased nuclear PI-PLC activity and cell cycle blockade in the G2 phase, suggesting a role for nuclear PI-PLC in the G2/M phase transition. Our data are consistent with the hypothesis that nuclear PI-PLC generates DAG to activate nuclear betaII PKC, whose activity is required for mitosis. PMID- 9334203 TI - The adjacent yeast genes ARO4 and HIS7 carry no intergenic region. AB - The region between the open reading frames of the adjacent yeast genes ARO4 and HIS7 consists of 417 base pairs (bp). Termination of ARO4 transcription and initiation of HIS7 transcription has to take place within this interval, because both genes are transcribed into the same direction. We show that the ARO4 terminator and the HIS7 promoter are spatially separated, nonoverlapping units. The ARO4 terminator includes 84 bp of the ARO4 3'-untranslated region with several redundant ARO4 3' end processing signals. Deletion of the ARO4 terminator does reduce but not completely shut down its expression. The adjacent region of 40 bp is neither required for correct ARO4 3' end formation nor for HIS7 initiation but contains the nucleotides corresponding to the wild type mRNA 3' ends. The following 280 bp are required for the HIS7 promoter. Replacement of the housekeeping ARO4 promoter by the stronger ACT1 promoter leads to reduced HIS7 expression due to transcriptional interference. This underlines the compactness of the yeast genome carrying virtually no intergenic regions between adjacent genes. PMID- 9334204 TI - Inhibition of ultraviolet B-induced activator protein-1 (AP-1) activity by aspirin in AP-1-luciferase transgenic mice. AB - Aspirin is under consideration as a promising chemopreventative agent for human cancers. To study the usefulness of aspirin as a chemopreventative agent for UV induced human skin cancer, we investigated the effect of aspirin on UVB-induced activator protein-1 (AP-1) activity. In the JB6 cell culture system, aspirin or sodium salicylate (SA) inhibited UVB-induced AP-1 activity in a dose-dependent manner; this inhibitory effect occurred only in cells pretreated with aspirin or SA before UVB irradiation but not cells treated with aspirin or SA after UVB irradiation. Furthermore, these inhibitory effects on UVB-induced AP-1 activity appeared to be mediated through blocking of activation of MAP kinase family members, including extracellular signal-regulated protein kinases, c-Jun N terminal kinases, and p38. It was not due to absorption of UVB light by aspirin. In the skin of AP-1-luciferase transgenic mice, UVB irradiation induced a rapid increase in AP-1 activity, which reached the peak at 48 h post-UVB irradiation. The topical pretreatment of mouse skin with aspirin markedly blocked the UVB induced AP-1 transactivation in vivo. These data provide the first evidence that aspirin and SA are inhibitors of UV-induced signal transduction and thus could be used as a chemopreventative agent for skin cancer. PMID- 9334205 TI - Caffeine and halothane sensitivity of intracellular Ca2+ release is altered by 15 calcium release channel (ryanodine receptor) mutations associated with malignant hyperthermia and/or central core disease. AB - Malignant hyperthermia (MH) and central core disease (CCD) are autosomal dominant disorders of skeletal muscle in which a potentially fatal hypermetabolic crisis can be triggered by commonly used anesthetic agents. To date, 17 mutations in the human RYR1 gene encoding the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (the ryanodine receptor) have been associated with MH and/or CCD. Although many of these mutations have been linked to MH and/or CCD, with high lod (log of the odds favoring linkage versus nonlinkage) scores, others have been found in single, small families. Independent biochemical evidence for a causal role for these mutations in MH is available for only two mutants. Mutations corresponding to the human MH mutations were made in a full-length rabbit RYR1 cDNA, and wild type and mutant cDNAs were transfected into HEK-293 cells. After about 48 h, intact cells were loaded with the fluorescent Ca2+ indicator, fura-2, and intracellular Ca2+ release, induced by caffeine or halothane, was measured by photometry. Ca2+ release in cells expressing MH or CCD mutant ryanodine receptors was invariably significantly more sensitive to low concentrations of caffeine and halothane than Ca2+ release in cells expressing wild type receptors or receptors mutated in other regions of the molecule. Linear regression analysis showed that there is a strong correlation (r = 0.95, p < 0.001) between caffeine sensitivity of different RYR1 mutants measured by the cellular Ca2+ photometry assay and by the clinical in vitro caffeine halothane contracture test (IVCT). The correlation was weaker, however, for halothane (r = 0.49, p > 0.05). Abnormal sensitivity in the Ca2+ photometry assay provides supporting evidence for a causal role in MH for each of 15 single amino acid mutations in the ryanodine receptor. The study demonstrates the usefulness of the cellular Ca2+ photometry assay in the assessment of the sensitivity to caffeine and halothane of specific ryanodine receptor mutants. PMID- 9334206 TI - Organic anion transporting polypeptide mediates organic anion/HCO3- exchange. AB - Organic anion transporting polypeptide (oatp) is an integral membrane protein cloned from rat liver that mediates Na+-independent transport of organic anions such as sulfobromophthalein and taurocholic acid. Previous studies in rat hepatocytes suggested that organic anion uptake is associated with base exchange. To better characterize the mechanism of oatp-mediated organic anion uptake, we examined transport of taurocholate in a HeLa cell line stably transfected with oatp under the regulation of a zinc-inducible promoter (Shi, X., Bai, S., Ford, A. C., Burk, R. D., Jacquemin, E., Hagenbuch, B., Meier, P. J., and Wolkoff, A. W. (1995) J. Biol. Chem. 270, 25591-25595). Whereas noninduced transfected cells showed virtually no uptake of [3H]taurocholate, taurocholate uptake by induced cells was Na+-independent and saturable (Km = 19.4 +/- 3.3 microM; Vmax = 62.2 +/ 1.4 pmol/min/mg protein; n = 3). To test whether organic anion transport is coupled to HCO3- extrusion, we compared the rates of taurocholate-dependent HCO3- efflux from alkali-loaded noninduced and induced cells. Monolayers grown on glass coverslips were loaded with the pH-sensitive dye 2', 7'-bis(carboxyethyl)-5(6) carboxyfluorescein; intracellular pH (pHi) was measured by excitation ratio fluorometry. Noninduced and induced cells were alkalinized to an equivalent pHi ( approximately 7.7) by transient exposure to a 50 mM HCO3-, Cl--free solution. In the absence of extracellular Cl- and taurocholate, isohydric reduction of superfusate HCO3- concentration from 50 to 25 mM resulted in an insignificant change in pHi over time (dpHi/dt) in both groups. Addition of 25 microM taurocholate to the superfusate led to a rapid fall in pHi in induced (-0.037 +/- 0.011 pH units/min to pHi of 7.41 +/- 0.14) but not in noninduced (0.003 +/- 0.006 pH units/min to pHi of 7.61 +/- 0.08) cells (p < 0.03). These data indicate that oatp-mediated taurocholate transport is Na+-independent, saturable, and accompanied by HCO3- exchange. We conclude that organic anion/base exchange is an important, potentially regulatable component of oatp function. PMID- 9334207 TI - A new action of parathyroid hormone. receptor-mediated stimulation of extracellular acidification in human osteoblast-like SaOS-2 cells. AB - The major physiological function of parathyroid hormone (PTH) is the maintenance of Ca2+/Pi homeostasis via the parathyroid hormone/parathyroid hormone-related protein receptor (PTHR) in kidney and bone. An important consequence of PTHR activation in bone is enhanced local acidification of the extracellular space. Agonist activation of some seven transmembrane-domain receptors increases the extracellular acidification rate (ECAR). We utilized microphysiometry to investigate PTH-stimulated, receptor-mediated increases in ECAR in human osteoblast-like SaOS-2 cells. PTH-(1-34) elicited a large, acute, dose-dependent increase in ECAR with an EC50 of about 2 nM. The PTH-induced increase in ECAR was specific to cells expressing the PTHR and was inhibited by PTHR antagonists. Rapid, partial, homologous desensitization of the PTH-induced increase in ECAR was observed. Incubation of SaOS-2 cells with 8-bromo-cyclic AMP neither mimicked nor abrogated the PTH effect, and PTH stimulated an acute increase in ECAR in cAMP-resistant SaOS-2 Ca#4A cells. Stimulation of ECAR by PTH was independent of transient increases in cytosolic free calcium. Both inhibition and down regulation of PKC reduced the PTH-induced increase in ECAR. Inhibition of Na+/H+ exchange did not affect the PTH-induced ECAR response. We conclude that PTH caused a receptor-mediated, concentration-dependent, increase in ECAR, which was not dependent on the cAMP/PKA signaling pathway or the Na+/H+ exchanger but involved the action of PKC. Thus, acid production in bone, a physiologically important action of PTH, is not confined to osteoclasts as previously considered but is also mediated by osteoblasts. PMID- 9334208 TI - Phosphorylation-dependent reversible association of Ca2+/calmodulin-dependent protein kinase II with the postsynaptic densities. AB - The association of soluble Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) with postsynaptic densities (PSDs) was determined by activity assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting of the enzyme. Soluble CaM kinase II was autophosphorylated with ATP in the presence of Ca2+ and calmodulin, and then it was incubated with PSDs. Autophosphorylated CaM kinase II was precipitated with PSDs by centrifugation. The kinase that was not autophosphorylated did not precipitate with PSDs. These results indicate that the soluble previously autophosphorylated CaM kinase II associates with PSDs and forms PSD-CaM kinase II complex. A maximum of about 60 microg of soluble CaM kinase II bound to 1 mg of PSD protein under the experimental conditions. Ca2+ independent activity generated by autophosphorylation of the kinase was retained in the PSD-CaM kinase II complex. The CaM kinase II thus associated with PSDs phosphorylated a number of PSD proteins in both the absence and presence of Ca2+. When the CaM kinase II-PSD complex was incubated at 30 degrees C, its Ca2+ independent activity was gradually decreased. This decrease was correlated with dephosphorylation of the kinase and its release from PSD-CaM kinase II complex. These results indicate that CaM kinase II reversibly translocates to PSDs in a phosphorylation-dependent manner. PMID- 9334209 TI - Transcriptional repression mediated by the PR domain zinc finger gene RIZ. AB - The RIZ (G3B or MTB-Zf) zinc finger gene is structurally related to the myeloid leukemia gene, MDS1-EVI1, and the transcription repressor/differentiation factor, PRDI-BF1/BLIMP1, through a conserved amino-terminal motif, the PR domain. Similar to MDS1-EVI1, RIZ gene normally produces two protein products that differ by the PR domain. The smaller protein RIZ2 lacks the PR domain of RIZ1 but is otherwise identical to RIZ1. Here we show that RIZ proteins bind to GC-rich or Sp-1-binding elements and repress transcription. Both RIZ1 and RIZ2 repressed the herpes simplex virus thymidine kinase (HSV-TK) promoter, one of the best characterized eukaryotic promoters. Recombinant RIZ1 proteins were able to bind to HSV-TK promoter. This binding was mediated by the GC-rich Sp-1 elements of the promoter and the first three zinc finger motifs of RIZ1. RIZ also encodes a repressor domain that was mapped to the central region of the protein. Fusion of this region to the GAL4 DNA-binding domain generated GAL4 site-dependent transcriptional repressors. We also show that RIZ1 protein can efficiently repress the simian virus 40 (SV40) early promoter, which primarily consists of Sp 1 sites; RIZ2, however, only weakly repressed this promoter, suggesting a role for PR in modulating RIZ protein function. The data have implications for a role of RIZ proteins in the regulation of cellular gene promoters, many of which are characterized by GC-rich elements. PMID- 9334210 TI - Upstream stimulatory factor binding to the E-box at -65 is required for insulin regulation of the fatty acid synthase promoter. AB - Fatty acid synthase (FAS) plays a central role in de novo lipogenesis in mammals. We have shown that FAS transcription rate is induced dramatically when fasted animals are refed with a high carbohydrate diet or when streptozotocin-diabetic mice are given insulin. We also reported that FAS gene transcription was up regulated by insulin through the proximal promoter region from -71 to -50 and that upstream stimulatory factors (USFs), including USF1 and USF2, interact with this region in vitro. In the present study, by using site-directed mutagenesis of the -71/-50 region and correlating functional assays of the mutated promoter with USF binding activities, we demonstrate that the -65/-60 E-box motif (5'-CATGTG 3') is functionally required for insulin regulation and that USFs are in vivo components of the insulin response complex. Mutation of the -65/-60 E-box sequence abolished insulin response in both transiently and stably transfected 3T3-L1 adipocytes in the -2. 1 kb promoter context, which contains all the necessary regulatory elements of the promoter based on our previous transgenic mice studies, and in the minimal -67 promoter context. Gel mobility shift assays demonstrated that USFs can no longer bind to the -71/-50 promoter region when the E-box is mutated. Cotransfection of USF1 and USF2 expression vectors with the FAS promoter-luciferase reporter constructs increased insulin-stimulated FAS promoter activity. Moreover, cotransfection of dominant negative USF1 and USF2 mutants lacking the DNA binding domain inhibited the insulin stimulation of the FAS promoter activity. On the other hand, site-directed mutagenesis of the -65/-60 E box surrounding sequences within the overlapped tandem copies of sterol regulatory element-binding protein (SREBP) binding sites prevented SREBP from binding to -71/-50 promoter region in vitro but had no effect on insulin regulation of the FAS promoter in vivo. When rat liver nuclear extracts were used in gel mobility shift assays, only USF-containing protein-DNA complexes that can be supershifted by specific USF antibodies were observed. These results demonstrate that upstream stimulatory factor binding to the E-box at -65 is required for insulin regulation of the fatty acid synthase promoter. PMID- 9334211 TI - Differential modes of nuclear localization signal (NLS) recognition by three distinct classes of NLS receptors. AB - The targeting of karyophilic proteins to nuclear pores is mediated via the formation of a nuclear pore-targeting complex, through the interaction of nuclear localization signal (NLS) with its NLS receptor. Recently, a novel human protein, Qip1, was identified from a yeast two-hybrid system with DNA helicase Q1. This study demonstrates that Qip1 is a novel third class of NLS receptor that efficiently recognizes the NLS of the helicase Q1. Moreover, the data obtained in this study show that the specific interaction between Qip1 and the NLS of the helicase Q1 requires its upstream sequence of the minimal essential NLS. By using purified recombinant proteins alone in the digitonin-permeabilized cell-free transport system, it was demonstrated that the two known human NLS receptors, Rch1 and NPI-1, are able to transport all the tested NLS substrates into the nucleus, while Qip1 most efficiently transports the helicase Q1-NLS substrates, which contain its upstream sequence in so far as we have examined the system. Furthermore, in HeLa cell crude cytosol, it was found that endogenous Rch1 binds to all the tested NLS substrates, while the binding of endogenous NPI-1 is restricted to only some NLSs, despite the fact that NPI-1 itself shows binding activity to a variety of NLSs. These results indicate that at least three structurally and functionally distinct NLS receptors exist in the human single cell population, and suggest that the nuclear import of karyophilic proteins may be controlled in a complex manner at the NLS recognition step by the existence of a variety of NLS receptors with various specificities to each NLS. PMID- 9334212 TI - The role of mGrb10alpha in insulin-like growth factor I-mediated growth. AB - Several isoforms of Grb10 are known to interact with either the insulin receptor or the insulin-like growth factor I (IGF-I) receptor, or both. Inasmuch as the data in the literature on the function of Grb10 are not always concordant, we have investigated the role of one of these isoforms, mGrb10alpha, in cell proliferation. For this purpose, a plasmid expressing mGrb10alpha was stably transfected into p6 cells and other mouse embryo fibroblast cell lines. An overexpressed mGrb10alpha inhibits IGF-I-mediated growth, causes a delay in the S and G2 phases of the cell cycle, and partially reverses the transformed phenotype. In contrast, it has no effect on insulin stimulation of cell proliferation. These studies indicate that this isoform of Grb10 has an inhibitory effect on IGF-I signaling of cell proliferation. PMID- 9334213 TI - A region of the beta subunit of the interferon alpha receptor different from box 1 interacts with Jak1 and is sufficient to activate the Jak-Stat pathway and induce an antiviral state. AB - Coexpression of the alpha and betaL subunits of the human interferon alpha (IFNalpha) receptor is required for the induction of an antiviral state by human IFNalpha. To explore the role of the different domains of the betaL subunit in IFNalpha signaling, we coexpressed wild-type alpha subunit and truncated forms of the betaL chain in L-929 cells. Our results demonstrated that the first 82 amino acids (AAs) (AAs 265-346) of the cytoplasmic domain of the betaL chain are sufficient to activate the Jak-Stat pathway and trigger an antiviral state after IFNalpha2 binding to the receptor. This region of the betaL chain, required for Jak1 binding and activation, contains the Box 1 motif that is important for the interaction of some cytokine receptors with Jak kinases. However, using glutathione S-transferase fusion proteins containing amino- and carboxyl-terminal deletions of the betaL cytoplasmic domain, we demonstrate that the main Jak1 binding region (corresponding to AAs 300-346 on the beta subunit) is distinct from the Box 1 domain (AAs 287-295). PMID- 9334214 TI - Prothymosin alpha in vivo contains phosphorylated glutamic acid residues. AB - Human and monkey prothymosin alpha contain activated carbonyl groups on glutamic acid residues. Three lines of evidence indicate the existence of unusual phosphates. 1) Prothymosin alpha continued to be metabolically labeled with [32P]orthophosphoric acid despite a mutation at Ser1, the sole site of phosphate in purified bovine prothymosin alpha (Sburlati, A. R., De La Rosa, A., Batey, D. W., Kurys, G. L., Manrow, R. E., Pannell, L. K., Martin, B. M., Sheeley, D. M., and Berger, S. L. (1993) Biochemistry 32, 4587-4596). 2) Immediately upon cell lysis, the pH stability curves of metabolically labeled native [32P]prothymosin alpha or a [32P]histidine-tagged variant resembled the pH stability curve of acetyl phosphate. 3) After a brief incubation at pH 7, these curves changed from a pattern diagnostic for an acyl phosphate to that characteristic of a serine or threonine phosphate, an observation consistent with transfer of phosphate in vitro. Our data indicate that most of prothymosin alpha's phosphates are subject instantaneously to hydrolysis, based on the observation that greater than 90% of the phosphate initially found at pH 7 disappeared at the extremes of pH. Rapid loss of phosphate was not affected by the presence of phosphatase inhibitors including 50 mM sodium fluoride, 1 mM okadaic acid, and 0.5 mM calyculin A. The amount of phosphate missing could not be ascertained, but the trifling amount recovered on Ser or Thr depended heavily on conditions favoring the transient survival of labile phosphate. Further analysis using COS cells lysed in the presence of sodium borohydride showed that: 1) phosphate recovered on prothymosin alpha decreased 8-fold when lysates were treated with borohydride; 2) the reagent caused 4-8 glutamic acid residues/molecule to vanish; 3) using [3H]NaBH4, label was introduced into proline, a product derived from reductive cleavage of phosphoglutamate; and 4) [3H]proline was localized almost exclusively to a peptide with pronounced homology to the histone binding site of nucleoplasmin, a chromatin remodeling protein found in Xenopus laevis. Our data demonstrate that prothymosin alpha is energy-rich by virtue of stoichiometric amounts of glutamyl phosphate. PMID- 9334215 TI - Turnover of the acyl phosphates of human and murine prothymosin alpha in vivo. AB - Prothymosin alpha is a small, highly acidic, abundant, nuclear, mammalian protein which is essential for cell growth. Our laboratory has recently shown that primate prothymosin alpha contains stoichiometric amounts of phosphate on the glutamyl groups of the protein and that in vitro the phosphate undergoes rapid hydrolysis or transfer to a nearby serine residue. Here an assay for the presence of acyl phosphates in vivo has been developed by measuring stable phosphoserine and phosphothreonine in vitro. The assay was used to determine the half-life of the acyl phosphates on prothymosin alpha in vivo by pulse-labeling HeLa cells with [32P]orthophosphate and chasing using three different techniques: permeabilization with digitonin to allow extracellular ATP to equilibrate with the intracellular pool; electroporation in the presence of ATP to reduce the specific activity of [32P]ATP by expansion of the pool; and incubation with inorganic phosphate. Regardless of the method, the phosphate turned over with a half-life of 75-90 min. The ability of cells to phosphorylate old prothymosin alpha molecules was established by demonstrating equivalent labeling of the protein with [32P]orthophosphate in the presence and absence of cycloheximide. The half-life of the acyl phosphates was also studied in resting and growing NIH3T3 cells, with measured values of 30-35 and 70 min, respectively. Our data suggest that the "activity" of prothymosin alpha involves the turnover of its acyl phosphates and that it participates in a function common to all nucleated mammalian cells regardless of whether they are quiescent or undergoing rapid proliferation. This is the first measurement of the stability of protein-bound acyl phosphates in vivo. PMID- 9334216 TI - N-Ethylmaleimide-sensitive fusion protein contains high and low affinity ATP binding sites that are functionally distinct. AB - N-Ethylmaleimide-sensitive factor (NSF) has been shown to be involved in numerous intracellular membrane fusion events of both the regulated and constitutive secretory pathways. Sequence analysis indicates that the NSF subunit contains two nucleotide-binding sites, both with the classical Walker A and B motifs. In this report, we examine the nucleotide binding properties of NSF. The homotrimer contains three high affinity ATP-binding sites with Kd = 30-40 nM for ATP and Kd = 2 microM for ADP. This class of binding sites did not bind AMP, adenine, or GTP. A second class of lower affinity nucleotide binding sites with a Kd = 15-20 microM was also detected. Using various mutant forms of NSF, the high affinity nucleotide-binding sites were localized to the D2 domains and the low affinity sites were localized to the D1 domains. Functionally it is these lower affinity sites in D1 that are crucial for NSF activity. Nucleotide concentration greatly affected the ability of NSF to interact with alpha-SNAP.SNARE (soluble NSF attachment protein-SNAP receptor) complex, suggesting that only when the D1 domain ATP-binding sites are occupied does NSF bind to the alpha-SNAP.SNARE complex. PMID- 9334217 TI - alpha-KDOase activity in oyster and synthesis of alpha- and beta-4 methylumbelliferyl ketosides of 3-deoxy-D-manno-octulosonic acid. AB - Although alpha- and beta-linked 3-deoxy-D-manno-octulosonic acid (KDO) is found in lipopolysaccharides (LPSs) of Gram-negative bacteria, capsular polysaccharides of microorganisms, and plants, very little is known about its degradation. Using both thin-layer chromatography and the periodate-thiobarbituric acid reaction, we found that the hepatopancreas of oyster (Crassostrea virginica) contained an enzyme (alpha-KDOase) capable of releasing alpha-linked KDO from LPSs. To facilitate the studies of alpha-KDOase, we have carried out the synthesis of 4 methylumbelliferyl-alpha-KDO (alpha-KDO-MU) by conjugating the glycosyl chloride of the per-O-acetylated methylester of KDO with methylumbelliferone by the SN2 type reaction and the catalyzed phase-transfer. In both cases, the beta-anomer was obtained as the major product with a yield of about 80%, whereas the yield of alpha-anomer was only about 7%. Attempts to increase the yield of alpha-anomer were not successful. alpha-KDO-MU was used as substrate to follow the purification of alpha-KDOase from oyster hepatopancreas. The pH optimum for oyster alpha-KDOase was determined to be 4.5 using Re-LPS as substrate and 3.0 using alpha-KDO-MU as substrate. The enzyme was found to be stable in the pH range of 3-8. This enzyme released KDO from different LPSs, including Re-LPS from Escherichia coli and Salmonella minnesota, Rd-LPS from S. minnesota, and de-O acyl-Re-LPS (Kiang, J., Szu, S. C., Wang, L.X., Tang, M., and Lee, Y. C. (1997) Anal. Biochem. 245, 97-101). PMID- 9334218 TI - Expression and characterization of two pathogenic mutations in human electron transfer flavoprotein. AB - Defects in electron transfer flavoprotein (ETF) or its electron acceptor, electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO), cause the human inherited metabolic disease glutaric acidemia type II. In this disease, electron transfer from nine primary flavoprotein dehydrogenases to the main respiratory chain is impaired. Among these dehydrogenases are the four chain length-specific flavoprotein dehydrogenases of fatty acid beta-oxidation. In this investigation, two mutations in the alpha subunit that have been identified in patients were expressed in Escherichia coli. Of the two mutant alleles, alphaT266M and alphaG116R, the former is the most frequent mutation found in patients with ETF deficiency. The crystal structure of human ETF shows that alphaG116 lies in a hydrophobic pocket, under a contact residue of the alpha/beta subunit interface, and that the hydroxyl hydrogen of alphaT266 is hydrogen-bonded to N(5) of the FAD; the amide backbone hydrogen of alphaT266 is hydrogen-bonded to C(4)-O of the flavin prosthetic group (Roberts, D. L., Frerman, F. E. and Kim, J-J. P. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 14355-14360). Stable expression of the alphaG116R ETF required coexpression of the chaperonins, GroEL and GroES. alphaG116R ETF folds into a conformation different from the wild type, and is catalytically inactive in crude extracts. It is unstable and could not be extensively purified. The alphaT266M ETF was purified and characterized after stabilization to proteolysis in crude extracts. Although the global structure of this mutant protein is unchanged, its flavin environment is altered as indicated by absorption and circular dichroism spectroscopy and the kinetics of flavin release from the oxidized and reduced protein. The loss of the hydrogen bond at N(5) of the flavin and the altered flavin binding increase the thermodynamic stability of the flavin semiquinone by 10-fold relative to the semiquinone of wild type ETF. The mutation has relatively little effect on the reductive half reaction of ETF catalyzed by sarcosine and medium chain acyl-CoA dehydrogenases which reduce the flavin to the semiquinone. However, kcat/Km of ETF-QO in a coupled acyl-CoA:ubiquinone reductase assay with oxidized alphaT266M ETF as substrate is reduced 33-fold; this decrease is due in largest part to a decrease in the rate of disproportionation of the alphaT266M ETF semiquinone catalyzed by ETF-QO. PMID- 9334219 TI - Requirements for Epstein-Barr nuclear antigen 1 (EBNA1)-induced permanganate sensitivity of the epstein-barr virus latent origin of DNA replication. AB - Epstein-Barr nuclear antigen 1 (EBNA1) activates DNA replication from the Epstein Barr virus latent origin of DNA replication, oriP. EBNA1 binds cooperatively to four recognition sites in the dyad symmetry (DS) element of oriP, causing alterations in the origin DNA structure, which can be detected by the increased sensitivity of one Thy residue in two of the binding sites to permanganate oxidation. To better understand the significance of this EBNA1-induced origin distortion, we have investigated the DNA sequence and EBNA1 amino acid requirements for permanganate sensitivity. We have shown that the EBNA1 DNA binding and dimerization domains are sufficient to induce permanganate sensitivity and that amino acids 463-467, which form an extended chain that travels along the minor groove of the EBNA1 recognition site, play an important role in generating the DNA distortion. The EBNA1-induced permanganate sensitivity is independent of cooperative interactions between EBNA1 molecules on the origin and requires a specific sequence within the EBNA1 binding site. Using synthetic EBNA1 binding sites, we found that the inversion of a single AT base pair in the EBNA1 recognition sequence is sufficient to confer EBNA1-induced permanganate sensitivity. These studies indicate that permanganate oxidation can detect very minor alterations in DNA structure. PMID- 9334220 TI - Trapping of mammalian topoisomerase I and recombinations induced by damaged DNA containing nicks or gaps. Importance of DNA end phosphorylation and camptothecin effects. AB - We used purified mammalian topoisomerases I (top1) and oligonucleotides containing a unique top1 cleavage site to study top1-mediated cleavage and recombination in the presence of nicks and short gaps mimicking DNA damage. In general, top1 cleavage was not induced opposite to the nicks, and nicks upstream from the top1 cleavage site suppressed top1 activity. Irreversible top1 cleavage complexes ("suicide products" or "aborted complexes") were produced in DNA containing nicks or short gaps just opposite to the normal top1 cleavage site. Camptothecin enhanced the formation of the aborted top1 complexes only for nicks downstream from the cleavage site. These aborted (suicide) complexes can mediate DNA recombination and promote illegitimate recombination by catalyzing the ligation of nonhomologous DNA fragments (acceptors). We report for the first time that top1-mediated recombination is greatly enhanced by the presence of a phosphate at the 5' terminus of the top1 aborted complex (donor DNA). By contrast, phosphorylation of the 3' terminus of the gap did not affect recombination. At concentrations that strongly enhanced inhibition of intramolecular religation, resulting in an increase of top1 cleavable complexes, camptothecin did not reduce recombination (intermolecular religation). Nicks or gaps with 5'-phosphate termini would be expected to be produced directly by ionizing radiations or by processing of abasic sites and DNA lesions induced by carcinogens or drugs used in cancer chemotherapy. Thus, these results further demonstrate that DNA damage can efficiently trap top1-cleavable complexes and enhance top1-mediated DNA recombination. PMID- 9334221 TI - Sequence-specific interactions in the Tus-Ter complex and the effect of base pair substitutions on arrest of DNA replication in Escherichia coli. AB - Arrest of DNA replication in Escherichia coli is mediated by specific interactions between the Tus protein and terminator (Ter) sequences. Binding of Tus to a Ter site forms a asymmetric protein-DNA complex that arrests DNA replication in an orientation-dependent fashion. In this study, mutant Ter sites carrying single base pair substitutions at 16 different positions were examined for their ability to bind purified Tus protein and arrest DNA replication. In vitro competition assays demonstrated that base pair substitutions at positions 8 19 had significant effects on the free energy of Tus binding (DeltaDeltaG0 of 1.5 to >4. 0 kcal/mol). Concomitant with loss of binding affinity, mutations at these positions also showed significantly lower or undetectable replication arrest activities in vivo. Substitutions at positions 6, 20, and 21 had moderate effects on Tus-Ter interactions, suggesting that these base pairs contribute to, but are not absolutely critical for, Tus binding. Even though the effects on binding were minimal, these Ter mutants were not as efficient as wild type Tus-TerB complexes at arresting replication forks. Three new potential Ter sites, referred to as TerH, TerI, and TerJ, were identified by searching the E. coli genome for sequence similarity to a consensus Ter site sequence. PMID- 9334222 TI - Regulation of eIF-4E BP1 phosphorylation by mTOR. AB - The proteins eIF-4E BP1 and p70 S6 kinase each undergo an insulin/mitogen stimulated phosphorylation in situ that is partially inhibited by rapamycin. Previous work has established that the protein known as mTOR/RAFT-1/FRAP is the target through which the rapamycin.FKBP12 complex acts to dephosphorylate/deactivate the p70 S6 kinase; thus, some mTOR mutants that have lost the ability to bind to the rapamycin.FKBP12 complex in vitro can protect the p70 S6 kinase against rapamycin-induced dephosphorylation/deactivation in situ. We show herein that such mTOR mutants also protect eIF-4E BP1 against rapamycin induced dephosphorylation, and for both p70 S6 kinase and eIF-4E BP1, such protection requires that the rapamycin-resistant mTOR variant retains an active catalytic domain. In contrast, mutants of p70 S6 kinase rendered intrinsically resistant to inhibition by rapamycin in situ are not able to protect coexpressed eIF-4E BP1 from rapamycin-induced dephosphorylation. We conclude that mTOR is an upstream regulator of eIF-4E BP1 as well as the p70 S6 kinase; moreover, these two mTOR targets are regulated in a parallel rather than sequential manner. PMID- 9334223 TI - Zinc-induced Alzheimer's Abeta1-40 aggregation is mediated by conformational factors. AB - The heterogeneous precipitates of Abeta that accumulate in the brain cortex in Alzheimer's disease possess varying degrees of resistance to resolubilization. We previously found that Abeta1-40 is rapidly precipitated in vitro by physiological concentrations of zinc, a neurochemical that is highly abundant in brain compartments where Abeta is most likely to precipitate. We now present evidence that the zinc-induced precipitation of Abeta is mediated by a peptide dimer and favored by conditions that promote alpha-helical and diminish beta-sheet conformations. The manner in which the synthetic peptide is solubilized was critical to its behavior in vitro. Zinc-induced Abeta aggregation was dependent upon the presence of NaCl, was enhanced by alpha-helical-promoting solvents, but was abolished when the peptide stock solution was stored frozen. The Abeta aggregates induced by zinc were reversible by chelation, but could then be reprecipitated by zinc for several cycles, indicating that the peptide's conformation is probably preserved in the zinc-mediated assembly. In contrast, Abeta aggregates induced by low pH (5.5) were not resolubilized by returning the pH milieu to 7.4. The zinc-Abeta interaction exhibits features resembling the gelation process of zinc-mediated fibrin assembly, suggesting that, in events such as clot formation or injury, reversible Abeta assembly could be physiologically purposive. Such a mechanism is contemplated in the early evolution of diffuse plaques in Alzheimer's disease and suggests a possible therapeutic strategy for the resolubilization of some forms of Abeta deposit in the disease. PMID- 9334224 TI - Regulation of capacitative Ca2+ influx in human neutrophil granulocytes. Alterations in chronic granulomatous disease. AB - Ca2+ entry through the capacitative (store-regulated) pathway was shown to be inhibited in neutrophil granulocytes by the protein kinase C activator phorbol 12 myristate 13-acetate and the chemoattractant N-formyl-methionyl-leucyl phenylalanine (fMLP) by a hitherto unknown mechanism. Measuring both Ca2+ and Mn2+ entry into store-depleted cells we show in the present study that inhibition of the capacitative pathway is absent in various forms of chronic granulomatous disease. To establish the possible relationship between inhibition of the capacitative pathway and ability of O-2 production and consequent membrane depolarization, gradual changes of the membrane potential were evoked in neutrophils of healthy individuals. This was accomplished by pharmacological manipulation of the membrane potential and by variations of the concentration and type of the stimulant. Close relationship was observed between membrane depolarization and inhibition of Mn2+ entry through the capacitative transport route. Our results provide an explanation for the inhibitory action of fMLP and phorbol 12-myristate 13-acetate on capacitative cation influx and reveal that upon physiological stimulation, Ca2+ entry into neutrophils is restricted by the depolarization accompanying O-2 production. PMID- 9334225 TI - Topology mapping of the amino-terminal half of multidrug resistance-associated protein by epitope insertion and immunofluorescence. AB - The multidrug resistance-associated protein (MRP) is an integral membrane protein that causes multidrug resistance when overexpressed in mammalian cells. Within the ATP-binding cassette superfamily, MRP belongs to a subgroup of structurally and functionally related proteins that includes the yeast cadmium factor 1 and yeast oligomycin resistance I proteins, and the mammalian sulfonylurea receptors SUR1 and SUR2. Hydropathy analysis of these proteins predicts a unique membrane associated region at the amino terminus followed by a structural unit composed of 12 transmembrane (TM) domains and two nucleotide-binding domains that is characteristic of eukaryotic ATP-binding cassette transporters. The topology of the membrane-associated regions of MRP remains largely unknown and was investigated. Small hemagglutinin epitopes (YPYDVPDYAS) were inserted in predicted hydrophilic segments of the membrane-associated regions from the amino terminal half of MRP and these proteins were expressed in HeLa cells, and tested for their capacity to confer etoposide resistance. The polarity of the inserted tags with respect to plasma membrane was then deduced by immunofluorescence in intact and permeabilized cells. Insertion of epitopes at positions 4, 163, 271, 574, and 653 produced functional proteins while insertions at positions 127, 417, 461, and 529 abrogated the capacity of MRP to confer drug resistance. Epitopes inserted at positions 4, 163, and 574 were localized extracellularly, whereas those inserted at positions 271 and 653 revealed an intracellular location. Although a single epitope inserted at position 461 was compatible with MRP function, it was inaccessible to the anti-epitope antibody and two copies of the tag at that site abrogated MRP function. These results indicate that the amino terminus of MRP is extracellular, while the linker segment joining the first and second membrane-associated regions is intracellular as is the first nucleotide binding domain. Our findings are therefore consistent with a topological model of MRP that contains 5 TM segments in the first membrane-associated region and 6 TM segments in the second membrane region. PMID- 9334226 TI - Beta2-chimaerin is a high affinity receptor for the phorbol ester tumor promoters. AB - Beta2-chimaerin, a member of the GTPase-activating proteins for the small GTP binding protein p21Rac, possesses a single cysteine-rich domain with high homology to those implicated in phorbol ester and diacylglycerol binding in protein kinase C (PKC) isozymes. We have expressed beta2-chimaerin in Sf9 insect cells using the baculovirus expression system and determined that, like PKCs, beta2-chimaerin binds phorbol esters with high affinity in the presence of phosphatidylserine as a cofactor. Scatchard plot analysis using the radioligand [3H]phorbol 12,13-dibutyrate revealed a dissociation constant of 1.9 +/- 0.2 nM for beta2-chimaerin. Likewise, beta2-chimaerin is a high affinity receptor for the bryostatins, a class of atypical PKC activators. A detailed comparison of structure-activity relations using several phorbol ester analogs revealed striking differences in binding recognition between beta2-chimaerin and PKCalpha. Although the diacylglycerol 1-oleoyl-2-acetylglycerol binds with similar potency to both beta2-chimaerin and PKCalpha, the mezerein analog thymeleatoxin has 56 fold less affinity for binding to beta2-chimaerin. To establish whether beta2 chimaerin responds to phorbol esters in cellular systems, we overexpressed beta2 chimaerin in COS-7 cells and monitored its subcellular distribution after phorbol ester treatment. Interestingly, as described previously for PKC isozymes, beta2 chimaerin translocates from cytosolic to particulate fractions as a consequence of phorbol ester treatment. Our results demonstrate that beta2-chimaerin is a novel target for the phorbol ester tumor promoters. The expansion of the family of phorbol ester receptors strongly suggests a potential for the "non-kinase" receptors as cellular mediators of the phorbol ester responses. PMID- 9334227 TI - Characterization of an epithelial approximately 460-kDa protein that facilitates endocytosis of intrinsic factor-vitamin B12 and binds receptor-associated protein. AB - By using receptor-associated protein (RAP) as an affinity target, an intrinsic factor-vitamin B12 (IF-B12)-binding renal epithelial protein of approximately 460 kDa was copurified together with the transcobalamin-B12-binding 600-kDa receptor, megalin. IF-B12 affinity chromatography of renal cortex membrane from rabbit and man yielded the same approximately 460-kDa protein. Binding studies including surface plasmon resonance analyses of the protein demonstrated a calcium dependent and high affinity binding of IF-B12 to a site distinct from the RAP binding site. The high affinity binding of IF-B12 was dependent on complex formation with vitamin B12. Light and electron microscope autoradiography of rat renal cortex cryosections incubated directly with IF-57Co-B12 and rat proximal tubules microinjected in vivo with the radioligand demonstrated binding of the ligand to endocytic invaginations of proximal tubule membranes followed by endocytosis and targeting of vitamin B12 to lysosomes. Polyclonal antibodies recognizing the approximately 460-kDa receptor inhibited the uptake. Immunohistochemistry of kidney and intestine showed colocalization of the IF-B12 receptor and megalin in both tissues. In conclusion, we have identified the epithelial IF-B12-binding receptor as a approximately 460-kDa RAP-binding protein facilitating endocytosis. PMID- 9334228 TI - A cellular model for long QT syndrome. Trapping of heteromultimeric complexes consisting of truncated Kv1.1 potassium channel polypeptides and native Kv1.4 and Kv1.5 channels in the endoplasmic reticulum. AB - We demonstrated that overexpression of a cRNA encoding a truncated potassium channel polypeptide that contains the NH2 terminus and the first transmembrane segment (Kv1.1N206Tag) abolished the expression of Kv1.1 and Kv1.5 outward currents in Xenopus oocytes (Babila, T., Moscucci, A., Wang, H., Weaver, F. E. & Koren, G. (1994) Neuron 12, 615-626). Recently, we showed that expression of Kv1.1N206Tag in the heart of transgenic mice resulted in the creation of mice with prolongation of the surface electrocardiogram's QT interval (London, B., Han, X., Folco, E. & Koren, G. (1996) Biophys. J. 70, A2601). To study the dominant negative mechanism of Kv1.1N206Tag, we overexpressed it in GH3 cells, a pituitary cell line expressing Kv1. 5 and Kv1.4. RNase protection analysis comparing the steady-state levels of native Kv1.5 and Kv1.1N206Tag transcripts revealed an excess of Kv1.1N206Tag transcript. Immunoprecipitation analysis using 12CA5 monoclonal antibody detected a 25-kDa polypeptide in the transfected cells. The half-life of Kv1.1N206Tag was 2.6 h. Subcellular fractionation of cell lysates labeled with [35S]methionine revealed that Kv1.1N206Tag polypeptide is detectable in the particulate (membranous) fraction, but not in the soluble (cytosol) fraction. A series of double immunoprecipitations with 12CA5 and polyclonal antibodies against Kv1.5 and Kv1.4 revealed that Kv1.1N206Tag forms heteromultimeric complexes with the native Kv1.4 and Kv1.5 polypeptides. The steady-state levels of Kv1.5 were not affected by the overexpression of Kv1.1N206Tag. Immunofluorescence colocalization and confocal microscopy analyses revealed that Kv1.1N206TagFlag did not reach the plasma membrane, and its distribution pattern was characteristic to that of a resident endoplasmic reticulum polypeptide. Our observations establish that the negative effect of Kv1.1N206Tag is mediated by the formation of heteromultimeric complexes with the native channels and by the retention of these complexes in the endoplasmic reticulum. PMID- 9334229 TI - High affinity binding and allosteric regulation of Escherichia coli glycogen phosphorylase by the histidine phosphocarrier protein, HPr. AB - The histidine phosphocarrier protein (HPr) is an essential element in sugar transport by the bacterial phosphoenolpyruvate:sugar phosphotransferase system. Ligand fishing, using surface plasmon resonance, was used to show the binding of HPr to a nonphosphotransferase protein in extracts of Escherichia coli; the protein was subsequently identified as glycogen phosphorylase (GP). The high affinity (association constant approximately 10(8) M-1), species-specific interaction was also demonstrated in electrophoretic mobility shift experiments by polyacrylamide gel electrophoresis. Equilibrium ultracentrifugation analysis indicates that HPr allosterically regulates the oligomeric state of glycogen phosphorylase. HPr binding increases GP activity to 250% of the level in control assays. Kinetic analysis of coupled enzyme assays shows that the binding of HPr to GP causes a decrease in the Km for glycogen and an increase in the Vmax for phosphate, indicating a mixed type activation. The stimulatory effect of E. coli HPr on E. coli GP activity is species-specific, and the unphosphorylated form of HPr activates GP more than does the phosphorylated form. Replacement of specific amino acids in HPr results in reduced GP activation; HPr residues Arg-17, Lys-24, Lys-27, Lys-40, Ser-46, Gln-51, and Lys-72 were established to be important. This novel mechanism for the regulation of GP provides the first evidence directly linking E. coli HPr to the regulation of carbohydrate metabolism. PMID- 9334230 TI - Type VI collagen anchors endothelial basement membranes by interacting with type IV collagen. AB - Type VI collagen filaments are found associated with interstitial collagen fibers, around cells, and in contact with endothelial basement membranes. To identify type VI collagen binding proteins, the amino-terminal domains of the alpha1(VI) and alpha2(VI) chains and a part of the carboxyl-terminal domain of the alpha3(VI) chain were used as bait in a yeast two-hybrid system to screen a human placenta library. Eight persistently positive clones were identified, two coding the known matrix proteins fibronectin and basement membrane type IV collagen and the rest coding new proteins. The amino-terminal domain of alpha1(VI) was shown to interact with the carboxyl-terminal globular domain of type IV collagen. The specificity of this interaction was further studied using the yeast two-hybrid system in a one-on-one format and confirmed by using isolated protein domains in immunoprecipitation, affinity blots, and enzyme linked immunosorbent assay-based binding studies. Co-distribution of type VI and type IV collagens in human muscle was demonstrated using double labeling immunofluorescent microscopy and immunoelectron microscopy. The strong interaction of type VI collagen filaments with basement membrane collagen provided a possible molecular pathogenesis for the heritable disorder Bethlem myopathy. PMID- 9334231 TI - Binding of the catabolite repressor protein CcpA to its DNA target is regulated by phosphorylation of its corepressor HPr. AB - Catabolite repression of a number of catabolic operons in bacilli is mediated by the catabolite control protein CcpA, the phosphocarrier protein HPr from the phosphoenolpyruvate-dependent sugar transport system (PTS), and a cis-acting DNA sequence termed the catabolite-responsive element (cre). We present evidence that CcpA interacts with HPr that is phosphorylated at Ser46 (Ser(P) HPr) and that these proteins form a specific ternary complex with cre DNA. Titration experiments following the circular dichroism signal of the cre DNA indicate that this complex consists of two molecules of Ser(P) HPr, a CcpA dimer, and the cre sequence. Limited proteolysis experiments indicate that the domain structure of CcpA is similar to other members of the LacI/GalR family of helix-turn-helix proteins, comprised of a helix-turn-helix DNA domain and a C-terminal effector domain. NMR titration of Ser(P) HPr demonstrates that the isolated C-terminal domain of CcpA forms a specific complex with Ser(P) HPr but not with unphosphorylated HPr. Based upon perturbations to the NMR spectrum, we propose that the binding site of the C-terminal domain of CcpA on Ser(P) HPr forms a contiguous surface that encompasses both Ser(P)46 and His15, the site of phosphorylation by enzyme I of the PTS. This allows CcpA to recognize the phosphorylation state of HPr, effectively linking the process of sugar import via the PTS to catabolite repression in bacilli. PMID- 9334232 TI - Mapping peptide-binding domains of the human V1a vasopressin receptor with a photoactivatable linear peptide antagonist. AB - The study of antagonist-binding domains of the human V1a vasopressin receptor was performed using a radioiodinated photoreactive peptide antagonist. This ligand displayed a high affinity for the receptor expressed in Chinese hamster ovary cell membranes, and specifically labeled two protein bands with apparent molecular mass at 85-90 and 46 kDa. Our results clearly show that the V1a receptor is degraded during incubation with the ligand and that the 46-kDa species is probably the result of the 85-90-kDa species proteolytic cleavage. Truncation of the receptor was then confirmed by deglycosylation with N glycosidase F. A monoclonal antibody directed against a c-Myc epitope added at the receptor NH2 terminus allowed immunoprecipitation of the 85-90-kDa photolabeled species. The 46-kDa photolabeled protein never immunoprecipitated, indicating that the truncated form of the receptor lacks the NH2 terminus region. To localize photolabeled domains of the receptor, the 46-kDa protein was cleaved with V8 and/or Lys-C endoproteinases. The identity of the smallest photolabeled fragment, observed at approximately 6 kDa, was then confirmed by mutation of the potential V8 cleavage sites. Our results indicate that covalent labeling of the vasopressin V1a receptor with the photoreactive antagonist occurs in a region including transmembrane domain VII (residues Asn327-Lys370). PMID- 9334233 TI - The Na+/H+ antiporter potentiates growth and retinoic acid-induced differentiation of P19 embryonal carcinoma cells. AB - The Na+/H+ exchanger is a ubiquitous plasma membrane protein that is responsible for pH regulation and is activated by growth factors. We examined the role of the Na+/H+ exchanger in cell growth and differentiation. Treatment of P19 cells with the Na+/H+ exchanger inhibitor Hoe 694 eliminated retinoic acid-induced differentiation in this cell line. We developed a P19 embryonal carcinoma cell line that was deficient in the Na+/H+ antiporter. Na+/H+ exchanger-deficient cells were reduced in the rate of cell growth and this effect was enhanced by the removal of added HCO3- and by reducing extracellular pH. The antiporter-deficient cells were also markedly deficient in their ability to differentiate to neuronal like cells and recovered this ability when the Na+/H+ antiporter was reintroduced. The results show that the absence of Na+/H+ antiport as a pH regulatory mechanism can result in deficiencies in both cell growth and differentiation in embryonal carcinoma cells. PMID- 9334234 TI - Identification of the region in yeast S-II that defines species specificity in its interaction with RNA polymerase II. AB - Yeast S-II was found to stimulate yeast RNA polymerase II only and not mouse RNA polymerase II. To identify the molecular region of S-II that defines species specificity, we constructed six hybrid S-II molecules consisting of three regions from yeast and/or Ehrlich cell S-II and examined their activity in terms of RNA polymerase II specificity and suppression of 6-azauracil sensitivity in the yeast S-II null mutant. We found that the region 132-270 (amino acid positions) of yeast S-II is indispensable for specific interaction with yeast RNA polymerase II in vitro and for suppression of 6-azauracil sensitivity in vivo. The corresponding region of Ehrlich cell S-II, the region 132-262, was also shown to be essential for its interaction with mouse RNA polymerase II. This region is known to be less conserved than the N- and C-terminal regions in the S-II family suggesting that it is important in the interaction with transcription machinery proteins in a tissue and/or species-specific manner. PMID- 9334235 TI - Role of the cytoskeleton in calcium signaling in NIH 3T3 cells. An intact cytoskeleton is required for agonist-induced [Ca2+]i signaling, but not for capacitative calcium entry. AB - Treatment of NIH 3T3 cells with cytochalasin D (10 microM, 1 h at 37 degrees C) disrupted the actin cytoskeleton and changed the cells from a planar, extended morphology, to a rounded shape. Calcium mobilization by ATP or by platelet derived growth factor was abolished, while the ability of thapsigargin (2 microM) to empty calcium stores and activate calcium influx was unaffected. Similar experiments with nocodazole to depolymerize the tubulin network yielded identical results. Platelet-derived growth factor induced an increase in inositol phosphates, and this increase was undiminished in the presence of cytochalasin D. Therefore, the blockade of agonist responses by this drug does not result from decreased phospholipase C. Injection of inositol 1,4,5-trisphosphate (IP3) released calcium to the same extent in control and cytochalasin D-treated cells. Confocal microscopic studies revealed a significant rearrangement of the endoplasmic reticulum after cytochalasin D treatment. Thus, disruption of the cytoskeleton blocks agonist-elicited [Ca2+]i mobilization, but this effect does not result from a lower calcium storage capacity, impaired function of the IP3 receptor, or diminished phospholipase C activity. We suggest that cytoskeletal disruption alters the spatial relationship between phospholipase C and IP3 receptors, impairing phospholipase C-dependent calcium signaling. Capacitative calcium entry was not altered under these conditions, indicating that the coupling between depletion of intracellular calcium stores and calcium entry does not depend on a precise structural relationship between intracellular stores and plasma membrane calcium channels. PMID- 9334236 TI - DNA binding specificity of the CCAAT-binding factor CBF/NF-Y. AB - CBF is a heterotrimeric protein that binds to DNA containing CCAAT motifs. Here we have analyzed interactions of recombinant CBF with DNA using hydroxyl radical footprinting and methylation interference assays. In the CBF-DNA complex, three separate DNA regions are protected from hydroxyl radical cleavage, one located over and immediately adjacent to the CCAAT motif itself and the other two located on both sides of the CCAAT motif. The methylation interference assay showed, however, that only in the CCAAT motif region methylation of bases was able to interfere with the formation of a CBF-DNA complex, suggesting that CBF makes sequence-specific contacts only in the CCAAT motif region. To further determine the specific DNA sequences necessary for CBF binding, we employed a polymerase chain reaction-mediated random binding site selection method. This analysis showed that CBF binding to DNA requires the CCAAT sequence and other specific sequences immediately flanking both ends of the CCAAT motif. We also showed that the nature of the flanking nucleotide sequences affects the affinity of CBF for DNA. Interestingly, most of the CCAAT motifs present in various higher eukaryotic promoters correspond to the CBF binding sites that were selected, consistent with the hypothesis that these motifs are binding sites for CBF and, hence, that CBF could regulate transcription of numerous eukaryotic genes. PMID- 9334238 TI - Pairing of DNA fragments containing (GGA:TCC)n repeats and promotion by high mobility group protein 1 and histone H1. AB - Tandemly repeated DNA sequences of (GGA:TCC)n are found in tracts up to 50 base pairs long, dispersed at thousands of sites throughout the genomes of eukaryotes. Here we demonstrate the formation of complexes paired between two DNAs containing such repeats in vitro and show enhancement of the pairing by glutathione S transferase fusion proteins of high mobility group protein 1 and histone H1. This assembly depends on incubation time at 37 degrees C and concentrations of the proteins and DNA, and the enhancement is inhibited by distamycin and actinomycin D interacting DNA through the minor groove. Structure of the DNA-DNA complex is deduced by comparison of its mobility in gel electrophoresis with those of synthetic markers of heterotetramers. Three synthetic and genomic DNA fragments containing repeats that have different arrangements exhibit different efficiencies of DNA pairing, implying that the pairing is affected by the number of repeat units and the arrangement of repeats in a sequence. Intriguingly, pairing occurs between homologous fragments but not between heterologous DNAs among the three. These results suggest that the repeat-mediated DNA pairing plays a role in organization of higher order architecture of chromatin and possibly chromosome segregation requiring sequence-specific association events of DNA molecules. PMID- 9334237 TI - Individual beta cells within the intact islet differentially respond to glucose. AB - Insulin production by the pancreatic islet is tightly coupled to the concentration of blood glucose. The mechanism by which glucose controls proinsulin biosynthesis in beta cells is poorly understood. Analysis of insulin gene expression in individual cells within whole, living islets using adenovirus gene transfer and direct observation of insulin promoter-directed green fluorescent protein activity indicates that beta cells are functionally heterogeneous. An increase in glucose concentration not only stimulates expression within individual beta cells, but unexpectedly acts to increase the total number of positive cells. The net islet response to a given glucose stimulus reflects an integrated action of beta cells with individually differing behaviors. This additional level of functional complexity may provide new insights into the pathophysiology and treatment of diabetes mellitus. PMID- 9334239 TI - Transcriptional regulation of the human erythroid 5-aminolevulinate synthase gene. Identification of promoter elements and role of regulatory proteins. AB - We have characterized the 5'-flanking region of the human erythroid-specific 5 amino levulinate synthase (ALAS) gene (the ALAS2 gene) and shown that the first 300 base pairs of promoter sequence gives maximal expression in erythroid cells. Transcription factor binding sites clustered within this promoter sequence include GATA motifs and CACCC boxes, critical regulatory sequences of many erythroid cell-expressed genes. GATA sites at -126/-121 (on the noncoding strand) and -102/-97 were each recognized by GATA-1 protein in vitro using erythroid cell nuclear extracts. Promoter mutagenesis and transient expression assays in erythroid cells established that both GATA-1 binding sites were functional and exogenously expressed GATA-1 increased promoter activity through these sites in transactivation experiments. A noncanonical TATA sequence at the expected TATA box location (-30/-23) bound GATA-1- or TATA-binding protein (TBP) in vitro. Conversion of this sequence to a canonical TATA box reduced expression in erythroid cells, suggesting a specific role for GATA-1 at this site. However, expression was also markedly reduced when the -30/-23 sequence was converted to a consensus GATA-1 sequence (that did not bind TBP in vitro), suggesting that a functional interaction of both factors with this sequence is important. A sequence comprising two overlapping CACCC boxes at -59/-48 (on the noncoding strand) was demonstrated by mutagenesis to be functionally important. This CACCC sequence bound Sp1, erythroid Kruppel-like factor, and basic Kruppel-like factor in vitro, while in transactivation experiments erythroid Kruppel-like factor activated ALAS2 promoter expression through this sequence. A sequence at -49/-39 with a 9/11 match to the consensus for the erythroid specific factor NF-E2 was not functional. Promoter constructs with 5'-flanking sequence from 293 base pairs to 10.3 kilobase pairs expressed efficiently in COS-1 cells as well as in erythroid cells, indicating that an enhancer sequence located elsewhere or native chromatin structure may be required for the tissue-restricted expression of the gene in vivo. PMID- 9334240 TI - Involvement of caspase-1 and caspase-3 in the production and processing of mature human interleukin 18 in monocytic THP.1 cells. AB - Recently, human interleukin 18 (hIL-18) cDNA was cloned, and the recombinant protein with a tentatively assigned NH2-terminal amino acid sequence was generated. However, natural hIL-18 has not yet been isolated, and its cellular processing is therefore still unclear. To clarify this, we purified natural hIL 18 from the cytosolic extract of monocytic THP.1 cells. Natural hIL-18 exhibited a molecular mass of 18.2 kDa, and the NH2-terminal amino acid was Tyr37. Biological activities of the purified protein were identical to those of recombinant hIL-18 with respect to the enhancement of natural killer cell cytotoxicity and interferon-gamma production by human peripheral blood mononuclear cells. We also found two precursor hIL-18 (prohIL-18)-processing activities in the cytosol of THP.1 cells. These activities were blocked separately by the caspase inhibitors Ac-YVAD-CHO and Ac-DEVD-CHO. Further analyses of the partially purified enzymes revealed that one is caspase-1, which cleaves prohIL-18 at the Asp36-Tyr37 site to generate the mature hIL-18, and the other is caspase-3, which cleaves both precursor and mature hIL-18 at Asp71-Ser72 and Asp76-Asn77 to generate biologically inactive products. These results suggest that the production and processing of natural hIL-18 are regulated by two processing enzymes, caspase-1 and caspase-3, in THP.1 cells. PMID- 9334241 TI - The Groucho/transducin-like enhancer of split transcriptional repressors interact with the genetically defined amino-terminal silencing domain of histone H3. AB - Groucho is a transcriptional repressor implicated in Notch signaling and involved in neural development and segmentation in Drosophila. We are investigating the molecular mechanisms underlying the functions of Groucho and its mammalian homologs, the transducin-like Enhancer of split (TLE) proteins. We report that Groucho/TLEs are associated with chromatin in live cells and that they co-purify with isolated histones. Affinity chromatography and far Western blotting studies show further that native Groucho/TLE proteins interact specifically with histone H3 and not with other core histones. This interaction is mediated by the H3 amino terminal domain previously shown by genetic analysis in yeast to be essential for the role of H3 in transcriptional silencing. We also demonstrate that Groucho/TLEs form oligomeric structures in vivo. These combined findings suggest that transcription complexes containing Groucho/TLEs may associate with chromatin through interactions with the amino terminus of histone H3 and that these interactions may be propagated along the chromosome due to the ability of Groucho/TLEs to participate in higher order structures. PMID- 9334242 TI - Molecular characterization of a novel A kinase anchor protein from Drosophila melanogaster. AB - Activation of protein kinase A (PKA) at discrete intracellular sites facilitates oogenesis and development in Drosophila. Thus, PKA-anchor protein complexes may be involved in controlling these crucial biological processes. Evaluation of this proposition requires knowledge of PKA binding/targeting proteins in the fly. We now report the discovery and characterization of cDNAs encoding a novel, Drosophila A kinase anchor protein, DAKAP550. DAKAP550 is a large (>2300 amino acids) acidic protein that is maximally expressed in anterior tissues. It binds regulatory subunits (RII) of both mammalian and Drosophila PKAII isoforms. The tethering region of DAKAP550 includes two proximal, but non-contiguous RII binding sites (B1 and B2). The B1 domain (residues 1406-1425) binds RII approximately 20-fold more avidly than B2 (amino acids 1350-1369). Affinity purified anti-DAKAP550 IgGs were exploited to demonstrate that the anchor protein is expressed in many cells in nearly all tissues throughout the lifespan of the fly. However, DAKAP550 is highly enriched and asymmetrically positioned in subpopulations of neurons and in apical portions of cells in gut and trachea. The combination of RII (PKAII) binding activity with differential expression and polarized localization is consistent with a role for DAKAP550 in creating target loci for the reception of signals carried by cAMP. The DAKAP550 gene was mapped to the 4F1.2 region of the X chromosome; flies that carry a deletion for this portion of the X chromosome lack DAKAP550 protein. PMID- 9334243 TI - Regulation of clusterin gene expression by transforming growth factor beta. AB - Transforming growth factor beta (TGFbeta) induces the expression of a wide variety of genes in many cell types. Our previous studies have shown that TGFbeta stimulates both clusterin mRNA and protein levels, and induces its accumulation in the nucleus of CCL64 cells. To further investigate the molecular mechanism of clusterin mRNA induction by TGFbeta, we created a 1.3-kilobase rat clusterin promoter/luciferase reporter construct. We demonstrate that TGFbeta enhances luciferase activity 2.5-6-fold in transient transfection assays of epithelial, endothelial, and fibroblast cell lines. Deletional analysis reveals that an AP-1 binding site (5'-TGAGTCA) in the minimal promoter region is necessary for initiating transactivation by TGFbeta. A single T to G base mutation in the AP-1 site (5'-TGAGGCA) abolishes TGFbeta-induced clusterin promoter transactivation. In transcription factor decoy experiments, 23-mer oligonucleotides of wild type AP-1 reduce TGFbeta induction of clusterin mRNA levels and promoter transactivation, while an oligonucleotide containing the mutated AP-1 site has no effect. Two specific protein kinase C inhibitors, GF109203X and calphostin C, block TGFbeta-induced clusterin mRNA levels and promoter transactivation. Together these results indicate that TGFbeta regulates clusterin gene expression through an AP-1 site and its cognate transcription factor AP-1, and requires the involvement of protein kinase C. PMID- 9334244 TI - N2 fixation by Streptomyces thermoautotrophicus involves a molybdenum dinitrogenase and a manganese-superoxide oxidoreductase that couple N2 reduction to the oxidation of superoxide produced from O2 by a molybdenum-CO dehydrogenase. AB - N2 fixation by Streptomyces thermoautotrophicus follows the equation N2 + 4 12MgATP + 8H+ + 8e- --> 2NH3 + H2 + 4-12MgADP + 4-12Pi and exhibits features which are not obvious in the diazotrophic bacteria studied so far. The reaction is coupled to the oxidation of carbon monoxide (CO) by a molybdenum-containing CO dehydrogenase that transfers the electrons derived from CO oxidation to O2, thereby producing superoxide anion radicals (O-2). A manganese-containing superoxide oxidoreductase reoxidizes the O-2 anions to O2 and transfers the electrons to a MoFeS-dinitrogenase for the reduction of N2 to ammonium. Among the most striking properties of the S. thermoautotrophicus nitrogenase system are the dependence on O2 and O-2, the complete insensitivity of all components involved toward O2 and H2O2, the inability to reduce ethine or ethene, and a low MgATP requirement. In addition, the subunit structure of the S. thermoautotrophicus nitrogenase components and the polypeptides involved seem to be dissimilar from the known nitrogenases. PMID- 9334245 TI - Regulation of duodenal specific expression of the human adenosine deaminase gene. AB - Formation of the mammalian gastrointestinal tract is an ordered process of development and differentiation. Yet, the adult small intestine also retains the plasticity to respond to cues both internal and environmental to modulate intestinal function. The components that regulate this development, differentiation, and modulation at the molecular level are only now being elucidated. We have used the human adenosine deaminase (ADA) gene as a model to identify potential cis-regulatory components involved in these processes within the small intestine. In mammals, high levels of ADA in the small intestine are limited specifically to the differentiated enterocytes within the duodenal region. These studies describe the identification of a region of the human ADA gene, completely distinct from the previously identified T-cell enhancer, which is capable of directing the human intestinal expression pattern in the intestine of transgenic mice. The reporter gene expression pattern observed in these transgenic mice is identical to the endogenous gene along both the cephalocaudal and crypt/villus axis of development. Timing of this transgene activation, however, varies from that of the endogenous mouse gene in that the transgene is activated approximately 2 weeks earlier in development. Even so, this precocious activation is also limited to the epithelium of the developing villi strictly within the duodenal region of the small intestine. PMID- 9334246 TI - Smooth muscle cell phenotype-dependent transcriptional regulation of the alpha1 integrin gene. AB - The expressional regulation of chicken alpha1 integrin in smooth muscle cells was studied. The alpha1 integrin mRNA was expressed developmentally and was distributed dominantly in vascular and visceral smooth muscles in chick embryos. In a primary culture of smooth muscle cells, alpha1 integrin expression was dramatically down-regulated during serum-induced dedifferentiation. Promoter analyses revealed that the 5'-upstream region (-516 to +281) was sufficient for transcriptional activation in differentiated smooth muscle cells but not in dedifferentiated smooth muscle cells or chick embryo fibroblasts. Like other alpha integrin promoters, the promoter region of the alpha1 integrin gene lacks TATA and CCAAT boxes and contains binding sites for AP1 and AP2. The essential difference from other alpha integrin promoters is the presence of a CArG box-like motif. Deletion and site-directed mutation analyses revealed that the CArG box like motif was an essential cis-element for transcriptional activation in differentiated smooth muscle cells, whereas the binding sites for AP1 and AP2 were not. Using specific antibodies, a nuclear protein factor specifically bound to the CArG box-like motif was identified as serum response factor. These results indicate that alpha1 integrin expression in smooth muscle cells is regulated transcriptionally in a phenotype-dependent manner and that serum response factor binding plays a crucial role in this regulation. PMID- 9334248 TI - Evidence that the beta-isoform of the human glucocorticoid receptor does not act as a physiologically significant repressor. AB - Alternative splicing of the human glucocorticoid receptor (hGR) primary transcript generates two receptor isoforms, hGRalpha and hGRbeta, with different carboxyl termini diverging at amino acid 727. By reverse transcriptase-polymerase chain reactions it was previously demonstrated that the hGRbeta message had a widespread tissue distribution. To demonstrate the presence of hGRbeta as protein we produced specific rabbit antisera to hGRbeta, as well as a hGRbeta-specific mouse monoclonal IgM antibody, by peptide immunizations. By SDS-polyacrylamide gel electrophoresis and Western immunoblotting we showed that hGRbeta is endogenously expressed at the protein level in HeLa cells and human lymphatic leukemia cells. Using an antibody directed against an epitope shared by both isoforms we showed a relatively lower expression of the hGRbeta form. We also showed that hGRbeta bound to hsp90 by immunoprecipitation of in vitro translated hGRbeta in reticulocyte lysate with hsp90-specific antibodies, a coprecipitation occurring also in the presence of dexamethasone. We could not demonstrate that hGRbeta inhibited the effects of dexamethasone-activated hGRalpha on a glucocorticoid-responsive reporter gene. In conclusion, low hGRbeta expression levels and hGRbeta-hsp90 interaction maintained in the presence of ligand and lack of inhibition of hormone-activated hGRalpha effects challenge the concept of the hGRbeta isoform as a proposed dominant negative inhibitor of hGRalpha activity. PMID- 9334247 TI - Clostridium perfringens enterotoxin utilizes two structurally related membrane proteins as functional receptors in vivo. AB - Human and mouse cDNAs showing homology to the Clostridium perfringens enterotoxin (CPE) receptor gene (CPE-R) from Vero cells (DDBJ/EMBL/GenBankTM accession no. D88492) (Katahira, J., Inoue, N., Horiguchi, Y., Matsuda, M., and Sugimoto, N. (1997) J. Cell Biol. 136, 1239-1247) were cloned. They were classified into two groups, the Vero cell CPE receptor homologues and rat androgen withdrawal apoptosis protein (RVP1; accession no. M74067) homologues, based on the similarities of primary amino acid sequences. L929 cells that were originally insensitive to CPE became sensitive to CPE on their transfection with cDNAs encoding either the CPE receptor or RVP1 homologues, indicating that these gene products are not only structurally similar but also functionally active as receptors for CPE. By binding assay, the human RVP1 homologue showed differences in affinity and capacity of binding from those of the human CPE receptor. Northern blot analysis showed that mouse homologues of the CPE receptor and RVP1 are expressed abundantly in mouse small intestine. The expression of CPE-R mRNA in the small intestine was restricted to cryptic enterocytes, indicating that the CPE receptor is expressed in intestinal epithelial cells. These results are consistent with reports that CPE binds to the small intestinal cells via two different kinds of receptors. High levels of expression of CPE-R and/or RVP1 mRNA were also detected in other organs, including the lungs, liver, and kidneys, but only low levels were expressed in heart and skeletal muscles. These results indicate that CPE uses structurally related cellular proteins as functional receptors in vivo and that organs that have not so far been recognized as CPE sensitive have the potential to be targets of CPE. PMID- 9334249 TI - Absence of glucocorticoid receptor-beta in mice. AB - Two human glucocorticoid receptor (GR) isoforms, GRalpha and GRbeta, are derived from the same gene by alternative splicing involving exon 9 of the GR locus. The non-ligand binding isoform GRbeta was proposed to act as a transdominant negative inhibitor of GRalpha, thus modulating glucocorticoid responsiveness of target tissues. To study GRbeta in mice we characterized the genomic region around exon 9 of the murine GR gene. Sequence analysis revealed that the presumed exon 9beta contained an open reading frame of 59 amino acids. In contrast, human exon 9beta encoded only 15 amino acids. Using reverse transcriptase polymerase chain reaction the absence of GRbeta mRNA was demonstrated in all adult mouse tissues examined. To exclude the possibility that the polymerase chain reaction conditions employed were not suitable for the amplification of GRbeta mRNA, we synthesized an artificial template corresponding to the presumed GRbeta mRNA spanning exons 7, 8, and 9beta. Various amounts of this template were added to brain cDNA preparations and as little as 25 molecules were detectable under the polymerase chain reaction conditions chosen. Since GRbeta is not conserved across species its physiological significance in humans appears questionable. PMID- 9334250 TI - Structure of the 32-kDa galectin gene of the nematode Caenorhabditis elegans. AB - Galectins are a family of soluble beta-galactoside-binding lectins distributed in both vertebrates and invertebrates and, more recently, found also in fungus. The 32-kDa galectin isolated from the nematode Caenorhabditis elegans (Hirabayashi, J., Satoh, M., and Kasai, K. (1992) J. Biol. Chem. 267, 15485-15490) was the first "tandem repeat-type" galectin, containing two homologous carbohydrate binding sites. Here, we report the structure of the nematode 32-kDa galectin gene. Physical mapping by yeast artificial chromosome polytene filter hybridization revealed that the 32-kDa galectin gene is located on chromosome II. Analysis of the transcript (1.4 kilobases) showed the presence at its 5'-end of a 22-nucleotide trans-spliced leader sequence (SL1). The entire genomic structure spanning >5 kilobase pairs (kbp), including the 5'-noncoding region, two intervening sequences (introns 1 and 2), and the 3'-noncoding region, was completely determined by the combination of genomic polymerase chain reaction and conventional colony hybridization. Intron 1 was relatively long (2.4 kbp) and was found to be inserted after the ninth codon (TAC) from the initiation codon. This position proved to be almost homologous to the conserved first intron insertion position in the vertebrate galectin genes (i. e. genes of mammalian galectin-1, 2, and -3 and chick 14-kDa galectin). On the other hand, intron 2 was much shorter (0.6 kbp), and it was inserted into the central region of the second carbohydrate-binding site. Although such an insertion pattern has never been observed in the vertebrate galectin genes, it seems to be common in C. elegans tandem repeat-type galectin genes, as predicted by the C. elegans genome project (Coulson, A., and the C. elegans Genome Consortium (1996) Biochem. Soc. Trans. 24, 289-291). Based on extensive sequence comparison, the origin and molecular evolution of the tandem repeat-type galectins are discussed. PMID- 9334251 TI - MUC3 human intestinal mucin. Analysis of gene structure, the carboxyl terminus, and a novel upstream repetitive region. AB - MUC3 is a large mucin glycoprotein expressed by the human intestine and gall bladder. In this manuscript, we present details of the deduced protein structure of MUC3. The MUC3 carboxyl-terminal domain is 617 residues in length, including 511 residues of a non-repetitive mucin-like domain (27% Thr, 22% Ser, and 11% Pro) and a 106-residue Cys-rich domain with homology to the epidermal growth factor (EGF) -like structural motifs found in many proteins. The region of MUC3 located upstream of the previously described 51-base pair (bp) tandem repeats, which encode a major Ser and Thr-rich domain, consists of a second type of repetitive structure with an imperfect periodicity of approximately 1125 bp. This domain is also mucin-like and appears to be considerably larger than 2000 residues (6000 bp). The MUC3 gene itself is large and complex. Using pulse field gel electrophoresis and blot analysis, the smallest fragment found that contained all human genomic DNA hybridizing to the 51-bp tandem repeat probe was 200 kilobases with restriction enzyme SwaI. Both PvuII and PstI produced two sets of hybridizing fragments that were hypervariable within the human population with a pattern suggestive of both a variation in the number of tandem repeats (VNTR) and sequence polymorphism. These fragments varied independently of each other, but no genetic recombination was detected in a study of 40 human families. Thus, the MUC3 gene encodes a very large glycoprotein with a structure very different from that of any mucin currently described. PMID- 9334252 TI - C-Mannosylation of human RNase 2 is an intracellular process performed by a variety of cultured cells. AB - C2-alpha-Mannosyltryptophan was discovered in RNase 2 from human urine, representing a novel way of attaching carbohydrate to a protein. Here, we have addressed two questions related to the biosynthesis of this modification: (i) is C-mannosylation part of the normal intracellular biosynthetic route, and (ii) how general is it, i.e. which organisms perform this kind of glycosylation? To answer the first question, RNase 2, which is identical to the eosinophil-derived neurotoxin, was isolated from intracellular stores of cultured human HL-60 cells. The enzyme was C-mannosylated at Trp-7, showing that the modification occurs intracellularly, before secretion of the protein. The second question was investigated by immunological and chemical analysis of RNase 2 purified from the supernatant of transiently transformed cells from different organisms. This revealed that C-mannosylation occurs in cells from man, green monkey, pig, mouse, and hamster. The observation that pig kidney cells contain the machinery for C mannosylation of Trp-7 of human RNase 2 but that the homologous RNase from porcine kidney is not a substrate, since it does not contain a tryptophan at position 7, strongly suggests that C-mannosylated proteins other than RNase 2 exist. Recombinant RNase 2 isolated from insect cells, plant protoplasts, and Escherichia coli was not C-mannosylated. These results not only form the basis for further studies on the biochemical aspects of C-mannosylation but also have implications for the choice of cells for production of recombinant glycoproteins. PMID- 9334253 TI - Activation of three distinct RXR/RAR heterodimers induces growth arrest and differentiation of neuroblastoma cells. AB - Naturally occurring retinoids, like all-trans retinoic acid and 9-cis retinoic acid, are known to affect proliferation and differentiation of sensitive neuroblastoma cell lines. Cellular responsiveness to retinoic acid depends on its interaction with two distinct classes of receptors, the retinoic acid receptors (RARs) and the retinoic X receptors (RXRs). Both receptor classes have three different subtypes (RARalpha, RARbeta, and RARgamma and RXRalpha, RARbeta, and RARgamma) that act as ligand-dependent transcription factors. To examine the involvement of the different receptor classes and subtypes in the biological responses of neuroblastoma cells to retinoids, we analyzed the effects of a panel of receptor-selective retinoids on cell growth, differentiation, and gene expression on in vitro cultured KCNR cells. Any association of per se inactive RXR-selective with RAR-selective ligands efficiently regulates growth inhibition, differentiation (neurite extension), and expression of RARbeta, TrkB, and N-myc. SR11383 alone, a very potent retinoid, entirely reproduces the pattern of biological responses induced by naturally occurring retinoids. In contrast to other tumor cell lines, the growth of neuroblastoma cell lines is not altered using AP1-antagonistic retinoids. These studies raise the possibility that three distinct RXR/RAR heterodimers mediate the effects of retinoids on neuroblastoma cells through an AP-1 antagonism-independent mechanism. PMID- 9334254 TI - Functional analysis of the promoter for the human CYP1B1 gene. AB - Our laboratory has cloned the cDNA (Sutter, T. R., Tang, Y. M., Hayes, C. L., Wo, Y.-Y. P., Jabs, E. W., Li, X., Yin, H., Cody, C. W. , and Greenlee, W. F. (1994) J. Biol. Chem. 269, 13092-13099) and gene (Tang, Y. M., Wo, Y.-Y. P., Jabs, E. W., Stewart, J. C., Sutter, T. R., and Greenlee, W. F. (1996) J. Biol. Chem. 271, 28324-28330) for human CYP1B1, a new member of the cytochrome P450 superfamily. Here, we report on the mapping and function of the CYP1B1 promoter. The CYP1B1 promoter is fully functional, when it is uncoupled from upstream enhancer elements. Deletion analysis and site-directed mutagenesis identified four regulatory elements required for maximum promoter activity: two antisense Sp1 sites (-84 to -89 and -68 to -73), a TATA-like box (-34 to -29), and an initiator motif (-5 to +3). The initiator and the TATA-like elements are both required for basal promoter activity, with enhanced activity mediated by the two antisense Sp1 elements. The CYP1B1 initiator was demonstrated by in vitro transcription analysis to be a positioning element that maintained fidelity of transcription from a single site. Specific binding to a CYP1B1 initiator probe by human nuclear extract proteins was competed either by the highly homologous murine terminal deoxynucleotidyl transferase initiator or, to a lesser extent, by the adenovirus major late initiator. Taken together, these results indicate that the structure and function of the CYP1B1 promoter confers constitutive expression of the gene and assures fidelity of transcription initiation from a single site. The CYP1B1 promoter is distinct from the promoters of the closely related cytochrome P450s CYP1A1 and CYP1A2 and is structurally and functionally similar to the promoters of constitutively expressed genes and at least two viruses. PMID- 9334255 TI - Characterization of a 30-kDa peripheral nerve glycoprotein that binds laminin and heparin. AB - We have shown previously that a bovine peripheral nerve protein with a molecular mass of about 30 kDa binds laminin in blot overlay assay. In this paper, we have characterized this 30-kDa laminin-binding protein (LBP30). LBP30 was extracted from the crude bovine peripheral nerve membranes at pH 12 or by 0.5 M NaCl but not by 2% Triton X-100. LBP30 bound to heparin-Sepharose in the presence of 0.5 M NaCl. The results of lectin staining indicated that LBP30 contained both terminally sialylated and nonsialylated Ser/Thr-linked oligosaccharides. LBP30 bound laminin-2 as well as laminin-1 but not fibronectin or collagen type IV. When immobilized LBP30 was incubated with the crude peripheral nerve membrane extracts, all of the endogenous peripheral nerve laminin chain isoforms, the alpha1, alpha2, beta1, beta2, and gamma1 chains, were detected bound to LBP30. The binding of LBP30 to laminin was inhibited by heparin, heparan sulfate, dextran sulfate, or NaCl but was not affected significantly by chondroitin sulfate, dextran, or EDTA. Although LBP30 bound to laminin-1 denatured with SDS in a nonreducing condition, the binding was reduced drastically when laminin-1 was denatured with SDS in a reducing condition, suggesting that the binding of LBP30 is somewhat dependent on the high order structure of laminin-1. Immunohistochemical analysis demonstrated the broad distribution of LBP30 in the perineurium and endoneurium of bovine peripheral nerve. These results indicate that LBP30 is a laminin- and heparin-binding glycoprotein localized in the perineurium and endoneurium of bovine peripheral nerve. PMID- 9334256 TI - Widespread expression of chondroitin sulfate-type serglycins with CD44 binding ability in hematopoietic cells. AB - Serglycin is a family of small proteoglycans with Ser-Gly dipeptide repeats and is modified with various types of glycosaminoglycan side chains. We previously demonstrated that chondroitin sulfate-modified serglycin is a novel ligand for CD44 involved in the adherence and activation of lymphoid cells. In this study, we investigated the production and distribution of CD44 binding serglycins in various hematopoietic cells and characterized their carbohydrate side chains. Immunoprecipitation analysis using CD44-IgG and polyclonal antibody against the serglycin core peptide demonstrated that various serglycin species capable of binding CD44 are produced by a variety of hematopoietic cells including lymphoid cells, myeloid cells, and a few tumor cell lines. Glycosaminoglycans on these serglycins, which are essential for CD44 binding, are composed of chondroitin 4 sulfate or a mixture of chondroitin 4-sulfate and chondroitin 6-sulfate, but no heparin or heparan sulfate side chain was detected. The serglycins are also secreted by normal splenocytes, lymph node lymphocytes, and bone marrow cells, whereas they are secreted in very small amounts by normal thymocytes. Secretion of serglycins is greatly enhanced by mitogenic stimulation with concanavalin A or lipopolysaccharide. Our results showed that serglycin, unlike hyaluronate, is produced and secreted in a functional (CD44 binding) form by many members of the hematopoietic system including various lymphocyte subsets. Our data suggest that serglycin may serve as a major ligand for CD44 in various events in the lymphohematopoietic system. PMID- 9334257 TI - Identification of several human homologs of hamster DNA damage-inducible transcripts. Cloning and characterization of a novel UV-inducible cDNA that codes for a putative RNA-binding protein. AB - Low ratio hybridization subtraction technique was previously used in this laboratory to enrich and isolate a number of low abundance UV-inducible hamster transcripts (Fornace, A. J., Jr., Alamo, I. J., and Hollander, M. C. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 8800-8804) that led to the identification and cloning of five important hamster and human GADD genes (Fornace, A. J., Jr., Nebert, D. W., Hollander, M. C., Luethy, J. D., Papathanasiou, M., Fargnoli, J., and Holbrook, N. J. (1989) Mol. Cell. Biol. 9, 4196-4203). In this study we have characterized the remaining DNA damage-inducible (DDI) transcripts. Of the 24 DDI clones, 3 clones (A13, A20, and A113) representing different regions of the same hamster cDNA exhibited near perfect homology to human p21(WAF1/CIP1) cDNA. The DDI clones A26, A88, and A99 displayed very high sequence homologies with the human proliferating nuclear antigen, rat translation initiation factor-5 (eIF-5), and human thrombomodulin, respectively, whereas clones A29 and A121 matched with express sequence tagged sequences of unknown identity. The DDI clones A18, 106, and A107 were different isolates of the same hamster cDNA (hereafter referred to as A18) and displayed high sequence homology with the members in the heterogeneous ribonucleoprotein (hnRNP) family. Using the hamster A18 partial length cDNA as a probe, we screened human fibroblast cDNA library and isolated the corresponding full-length human cDNA. The deduced amino acid sequence revealed that the putative protein contains all the canonical features of a novel glycine-rich hnRNP. The A18 mRNA levels were specifically increased in response to DNA damage induced by UV irradiation or UV mimetic agents. Thus the putative A18 hnRNP is the first hnRNP whose mRNA is specifically regulated in response to UV-induced DNA damage; accordingly, it may play some role in repair of UV-type DNA damage. PMID- 9334258 TI - Sequence of cDNAs encoding components of vascular actin single-stranded DNA binding factor 2 establish identity to Puralpha and Purbeta. AB - Transcriptional repression of the mouse vascular smooth muscle alpha-actin gene in fibroblasts and myoblasts is mediated, in part, by the interaction of two single-stranded DNA binding activities with opposite strands of an essential transcription enhancer factor-1 recognition element (Sun, S., Stoflet, E. S., Cogan, J. G., Strauch, A. R., and Getz, M. J. (1995) Mol. Cell. Biol. 15, 2429 2436). One of these activities, previously designated vascular actin single stranded DNA-binding factor 2 includes two distinct polypeptides (p44 and p46) which specifically interact with the purine-rich strand of both the enhancer and a related element in a protein coding exon of the gene (Kelm, R. J., Jr., Sun, S., Strauch, A. R., and Getz, M. J. (1996) J. Biol. Chem. 271, 24278-24285). Expression screening of a mouse lung cDNA library with a vascular actin single stranded DNA-binding factor 2 recognition element has now resulted in the isolation of two distinct cDNA clones that encode p46 and p44. One of these proteins is identical to Puralpha, a retinoblastoma-binding protein previously implicated in both transcriptional activation and DNA replication. The other is a related family member, presumably Purbeta. Comparative band shift and Southwestern blot analyses conducted with cellular p46, p44, and cloned Pur proteins synthesized in vitro and in vivo, establish identity of p46 with Puralpha and p44 with Purbeta. This study implicates Puralpha and/or Purbeta in the control of vascular smooth muscle alpha-actin gene transcription. PMID- 9334259 TI - Hydroxyl radicals depolymerize glomerular heparan sulfate in vitro and in experimental nephrotic syndrome. AB - Heparan sulfate, the polysaccharide side chain of heparan sulfate proteoglycan, is important for the permselective properties of the glomerular basement membrane. In this report, we show a role for hydroxyl radicals in heparan sulfate degradation and an enhanced glomerular basement membrane permeability. First, in enzyme-linked immunosorbent assay, exposure of coated heparan sulfate (proteoglycan) to reactive oxygen species resulted in a +/-50% decrease of binding of a monoclonal antibody against heparan sulfate, whereas binding of an antibody against the core protein remained unaltered. Second, on polyacrylamide gel electrophoresis, the molecular weight of heparan sulfate exposed to radicals was reduced which indicates depolymerization. Both in enzyme-linked immunosorbent assay and gel electrophoresis, hydroxyl radicals are instrumental for heparan sulfate degradation as shown by the addition of various radical scavengers. Third, in an experimental model for human nephrotic syndrome (Adriamycin nephropathy in rats), glomerular basement membrane staining of two recently described anti-heparan sulfate antibodies (JM403 and KJ865) was reduced by 24 and 43%. Treatment of Adriamycin-exposed rats with the hydroxyl radical scavenger dimethylthiourea both reduced albuminuria by 37% (p < 0.01) and partly prevented loss of heparan sulfate staining by 53% (JM403) and 39% (KJ865) (p < 0.03). In contrast to the heparan sulfate side chains, the core protein expression and the extent of glycanation did not change in Adriamycin nephropathy. We conclude that glomerular basement membrane heparan sulfate is susceptible to depolymerization by hydroxyl radicals leading to loss of glomerular basement membrane integrity and albuminuria. PMID- 9334260 TI - African trypanosomes have differentially expressed genes encoding homologues of the Leishmania GP63 surface protease. AB - The genomes of various Leishmania parasites contain tandemly arrayed genes encoding an abundant 63-kDa surface glycoprotein called GP63. Leishmania GP63s are metalloproteases that play an important role in the invasion and survival of the parasites within the macrophage, and their presence on the Leishmania surface has been correlated with resistance to complement-mediated lysis. Here we report the identification of GP63-like genes in African trypanosomes. The predicted trypanosome and Leishmania GP63s share a metalloprotease catalytic site motif of HEXXH as well as 19 cysteines and 10 prolines, implying a conservation of enzymatic activity and secondary/tertiary structure. The trypanosome GP63 genes are transcribed equally in procyclic and bloodstream trypanosomes, but their mRNAs accumulate to a 50-fold higher steady state level in bloodstream trypanosomes, where the ratio of mRNAs for GP63 and variant surface glycoprotein is about 1:150. Transcription of the GP63 genes is sensitive to alpha-amanitin, indicating that they are transcribed by a different polymerase than the variant surface glycoprotein genes. These results lead to a reconsideration of the potential functions of GP63, inasmuch as African trypanosomes are not known to interact with macrophages and do not have an intracellular stage during their life cycle. PMID- 9334261 TI - The mouse homologue of the human transcription factor C1 (host cell factor). Conservation of forms and function. AB - The assembly of the herpes simplex virus (HSV) alpha/IE gene enhancer complex is determined by the interactions of the Oct-1 POU domain protein, the viral alphaTIF (alpha-trans-induction factor, VP16, ICP25, VMW65), and the C1 factor (host cell factor, HCF). A unique transcription factor, C1 consists of a family of polypeptides derived from a common precursor by site-specific proteolytic processing. To analyze the role of this factor in the determination of HSV lytic latent infection, cDNAs and genomic DNAs encoding the mouse homologue have been isolated. This factor is nearly identical to the human protein, contains multiple consensus proteolytic processing sites, and functions efficiently in the assembly of a specific HSV enhancer complex. Interestingly, the differential expression of the C1 factors in both human and mouse tissues may be important for the determination of HSV tissue tropism in these two organisms. PMID- 9334262 TI - Identification of an active site alanine in mevalonate kinase through characterization of a novel mutation in mevalonate kinase deficiency. AB - Sequencing of polymerase chain reaction-amplified cDNAs from cultured cells of three patients with mevalonate kinase deficiency revealed a G --> A transversion at nucleotide 1000 of the coding region, converting alanine to threonine at position 334 (A334T). To characterize this defect, we expressed wild-type and mutant cDNAs in Escherichia coli as the glutathione S-transferase fusion proteins, with purification by affinity chromatography. SDS-polyacrylamide gel electrophoresis analysis for wild-type and mutant fusion proteins indicated an expected molecular mass of 42-43 kDa. Kinetic characterization of the wild-type fusion protein yielded Km values of 150 +/- 23 and 440 +/- 190 microM (mean +/- S.E.) for substrates (RS)-mevalonate and ATP, respectively. Expressed wild-type mevalonate kinase (MKase) had a maximum velocity of 13.6 +/- 1.4 units/mg of protein (n = 22, +/-S.E.), whereas the A334T mutation yielded an enzyme with average Vmax of 0.26 +/- 0.02 unit/mg of protein (n = 6, +/-S.E.), representing a decrease to 1.4% of control Vmax. Restriction digestion with HhaI, in conjunction with direct sequencing of cDNAs, revealed that two patients were homozygous and one heterozygous for the A334T allele, establishing autosomal recessive inheritance within families. Although the A334T enzyme had a normal Km for ATP of 680 +/- 226 microM (n = 3, +/-S.E.), the Michaelis constant for (RS)-mevalonate was increased >30-fold to 4623 +/- 1167 microM (n = 4, +/-S.E.) under standard assay conditions. Comparable kinetic results were obtained using extracts of lymphoblasts, which were homozygous for the A334T allele. Alanine 334 is invariant in MKase from bacteria to man and located in a glycine-rich region postulated to have homology with ATP-binding sequences. Our results indicate that the bacterial expression system for human MKase will provide a useful model system in which to analyze inherited mutations and identify the first active site residue in MKase associated with stabilization of mevalonate binding. PMID- 9334263 TI - A novel juxtamembrane domain isoform of HER4/ErbB4. Isoform-specific tissue distribution and differential processing in response to phorbol ester. AB - Human epidermal growth factor receptor 4 (HER4) is a member of the epidermal growth factor (EGF) receptor subfamily of receptor tyrosine kinases that is activated by neuregulins (NRG), betacellulin (BTC), and heparin-binding EGF-like growth factor. Sequencing of full-length human HER4 cDNAs revealed the existence of two HER4 isoforms that differed by insertion of either 23 or 13 alternative amino acids in the extracellular juxtamembrane (JM) region. The 23-amino acid form (HER4 JM-a) and the 13-amino acid form (HER4 JM-b) were expressed in a tissue-specific manner, as demonstrated by reverse transcriptase-polymerase chain reaction analysis of mouse and human tissues. Both isoforms were expressed in neural tissues such as cerebellum, whereas kidney expressed HER4 JM-a only and heart HER4 JM-b only. In situ hybridization using specific oligonucleotides demonstrated transcription of both JM-a and JM-b isoforms in the mouse cerebellum. Tyrosine phosphorylation analysis indicated that both receptor isoforms were activated to the same extent by NRG-beta1 and BTC, and to a lesser extent by NRG-alpha1 and heparin-binding EGF-like growth factor. A functional difference was found, however, in response to phorbol ester treatment. Stimulation of cells with phorbol ester resulted in a loss of 125I-NRG-beta1 binding and in a reduction of total cell-associated HER4 protein in HER4 JM-a transfectants but not in HER4 JM-b transfectants. It was concluded that novel alternatively spliced isoforms of HER4 exist, that they are distributed differentially in vivo in mouse and human tissues, that they are both activated by HER4 ligands, and that they may represent cleavable and noncleavable forms of HER4. PMID- 9334264 TI - RNA ligands selected by cleavage stimulation factor contain distinct sequence motifs that function as downstream elements in 3'-end processing of pre-mRNA. AB - Critical events in 3'-end processing of pre-mRNA are the recognition of the AAUAAA polyadenylation signal by cleavage and polyadenylation specificity factor (CPSF) and the binding of cleavage stimulation factor (CstF) via its 64-kDa subunit to the downstream element. The stability of this CPSF.CstF.RNA complex is thought to determine the efficiency of 3'-end processing. Since downstream elements reveal high sequence variability, in vitro selection experiments with highly purified CstF were performed to investigate the sequence requirements for CstF-RNA interaction. CstF was purified from calf thymus and from HeLa cells. Surprisingly, calf thymus CstF contained an additional, novel form of the 64-kDa subunit with a molecular mass of 70 kDa. RNA ligands selected by HeLa and calf thymus CstF contained three highly conserved sequence elements as follows: element 1 (AUGCGUUCCUCGUCC) and two closely related elements, element 2a (YGUGUYN0-4UUYAYUGYGU) and element 2b (UUGYUN0-4AUUUACU(U/G)N0-2YCU). All selected sequences tested functioned as downstream elements in 3'-end processing in vitro. A computer survey of the EMBL data library revealed significant homologies to all selected elements in naturally occurring 3'-untranslated regions. The majority of element 2a homologies was found downstream of coding sequences. Therefore, we postulate that this element represents a novel consensus sequence for downstream elements in 3'-end processing of pre-mRNA. PMID- 9334266 TI - V1-situated stalk subunits of the yeast vacuolar proton-translocating ATPase. AB - The proton-translocating ATPase of the yeast vacuole is an enzyme complex consisting of a large peripheral membrane sector (V1) and an integral membrane sector (V0), each composed of multiple subunits. The V1 sector contains subunits that hydrolyze ATP, whereas the V0 sector contains subunits that translocate protons across the membrane. Additional subunits in both sectors couple these activities. Here we have continued our examination of intermediate subunits primarily associated with the V1 but also implicated in interactions with the V0. Interactions between Vma7p (F) and Vma8p (D) and between Vma4p (E) and Vma10p (G) are described. Although Vma7p and Vma10p have been observed to interact with the V0 sector, our results indicate that these subunits behave primarily as canonical V1 sector subunits. We categorize these four subunits as "stalk" subunits to distinguish them from the known catalytic (A and B) and proton-translocating (c, c', and Vma16p) subunits and to highlight their intermediate nature. Furthermore, we show that the in vivo stability of Vma4p is dependent upon interaction with Vma10p. This may be important in the regulation of assembly, since these two subunits add to the V1 during later stages of V1 assembly. This is the first demonstration of interdependence between ATPase subunits for structural stability. PMID- 9334265 TI - A new member of the cationic amino acid transporter family is preferentially expressed in adult mouse brain. AB - We have isolated and characterized a novel member (CAT3) of the cationic amino acid transporter (CAT) family. In oocyte injection assays, CAT3 cRNA exhibited a saturable, sodium ion-independent transport activity with high affinity for L arginine and L-lysine (Km = 40-60 and 115-165 microM, respectively). Transport of L-arginine was effectively competed only by cationic amino acids in L-form: arginine, lysine, ornithine, and 2,4-diamino-n-butyric acid but not by 2,3 diaminopropionic acid. The presence of L-arginine in the incubation medium stimulated the efflux rate of L-arginine, indicating that CAT3 is subject to trans-stimulation. All these results are consistent with the idea that CAT3, along with CAT1 and CAT2, constitutes the transport activity originally assigned to system y+. Like CAT2, but unlike CAT1, the expression of CAT3 is regulated in a highly tissue-specific manner; when various adult tissues were examined, significant levels of CAT3 transcript were detectable only in brain. In situ hybridization on brain sections revealed that CAT3 transcripts were localized predominantly along the midbrain-thalamus-hypothalamus axis, whereas neither CAT1 nor CAT2 transcripts demonstrated a similar localization. In contrast to its highly localized expression during the primitive streak stage and in the adult stage, CAT3 expression was detected more widely in 13.5 day post-coitum mouse embryos. PMID- 9334267 TI - Patterning the Xenopus blastula. AB - This review starts from the classical standpoint that there are at least two separable processes acting with respect to axis formation and tissue specification in the early Xenopus embryo: a UV-insensitive event establishing a postgastrula embryo consisting of three concentric germ layers, ectoderm, mesoderm and endoderm, all of a ventral character; and a UV-sensitive event producing tissue of a dorsal type, including somites, notochord and neural tissue, and concomitantly establishing the dorsoventral and anteroposterior axes. The experimental evidence suggesting the molecular basis of the dorsal and ventral pathways is reviewed. PMID- 9334268 TI - The expression of the C. elegans labial-like Hox gene ceh-13 during early embryogenesis relies on cell fate and on anteroposterior cell polarity. AB - Clusters of homeobox-containing HOM-C/hox genes determine the morphology of animal body plans and body parts and are thought to mediate positional information. Here, we describe the onset of embryonic expression of ceh-13, the Caenorhabditis elegans orthologue of the Drosophila labial gene, which is the earliest gene of the C. elegans Hox gene cluster to be activated in C. elegans development. At the beginning of gastrulation, ceh-13 is asymmetrically expressed in posterior daughters of anteroposterior divisions, first in the posterior daughter of the intestinal precursor cell E and then in all posterior daughters of the AB descendants ABxxx. In this paper, we present evidence that supports position-independent activation of ceh-13 during early C. elegans embryogenesis, which integrates cell fate determinants and cell polarity cues. Our findings imply that mechanisms other than cell-extrinsic anteroposterior positional signals play an important role in the activation and regulation of the C. elegans Hox gene ceh-13. PMID- 9334269 TI - meander tail acts intrinsic to granule cell precursors to disrupt cerebellar development: analysis of meander tail chimeric mice. AB - The murine mutation meander tail (gene symbol: mea) causes a near-total depletion of granule cells in the anterior lobe of the cerebellum, as well as aberrantly located Purkinje cells with misoriented dendrites and radial glia with stunted processes. Whether one, two or all three of these cell types is the primary cellular target(s) of the mutant gene is unknown. This issue is addressed by examining cerebella from adult chimeras in which both the genotype and phenotype of individual cells are marked and examined. From this analysis, three novel observations are made. First, genotypically mea/mea Purkinje cells and glial cells exhibit normal morphologies in the cerebella of chimeric mice indicating that the mea gene acts extrinsically to these two cell populations. Second, few genotypically mea/mea granule cells are present in the anterior lobe or, unexpectedly, in the posterior lobe. These findings indicate that the mea gene acts intrinsically to the granule cell or its precursors to perturb their development. Third, there are near-normal numbers of cerebellar granule cells in the chimeric cerebellum. This result suggests that mea/mea cells are out-competed and subsequently replaced by an increased cohort of wild-type granule cells resulting from an upregulation of wild-type granule cells in the chimeric environment. We propose that the wild-type allele of the mea gene is critical for the developmental progression of the early granule cell neuroblast. PMID- 9334271 TI - Hoxb-8 has a role in establishing early anterior-posterior polarity in chick forelimb but not hindlimb. AB - The distribution of Hoxb-8 transcripts through the chick flank and early forelimb mirrors the distribution of polarizing activity in the flank at these early stages. Polarizing activity displayed by Hoxb-8-expressing tissue is only realised when placed adjacent to the AER and appears to be mediated through Shh induction, suggesting that Hoxb-8 may lie genetically upstream of Shh. Accordingly, Hoxb-8 expression is rapidly induced by retinoic acid (RA) treatment in the anterior of the forelimb in a spatial and temporal manner that is consistent with the induction of Shh and formation of the ZPA. Furthermore, inhibition of RA synthesis in the flank downregulates the expression of endogenous Hoxb-8 and results in the loss of Shh expression. However, once the ZPA has become established the posterior limb mesoderm displays resistance to the induction of Hoxb-8 expression. Grafting of ZPA cells to the anterior of a host limb renders the host anterior tissue resistant to RA-induced Hoxb-8 expression. These results indicate that Hoxb-8 expression may be regulated by the established ZPA through a negative feedback loop. The anterior AER also secretes an inhibitory factor, preventing RA-induced or already established Hoxb-8 expression in the cells immediately underneath the AER. Consistent with a role for Hoxb-8 in positioning of the forelimb ZPA, Hoxb-8 expression is not seen in RA-induced duplications at the anterior of the hindlimb. However, grafting of Hoxb-8 expressing tissue to the hindlimb can lead to Shh expression and similar duplications, suggesting that factors mediating ZPA formation are very similar in both wing and leg. PMID- 9334270 TI - Differentiation of engrafted multipotent neural progenitors towards replacement of missing granule neurons in meander tail cerebellum may help determine the locus of mutant gene action. AB - Previously we observed that stable clones of multipotent neural progenitor cells, initially isolated and propagated from the external granular layer of newborn wild-type mouse cerebellum, could participate appropriately in cerebellar development when reimplanted into the external granular layer of normal mice. Donor cells could reintegrate and differentiate into neurons (including granule cells) and/or glia consistent with their site of engraftment. These findings suggested that progenitors might be useful for cellular replacement in models of aberrant neural development or neurodegeneration. We tested this hypothesis by implanting clonally related multipotent progenitors into the external granular layer of newborn meander tail mice (gene symbol=mea). mea is an autosomal recessive mutation characterized principally by the failure of granule cells to develop in the cerebellar anterior lobe; the mechanism is unknown. We report that approximately 75% of progenitors transplanted into the granuloprival anterior lobe of neonatal mea mutants differentiated into granule cells, partially replacing or augmenting that largely absent neuronal population in the internal granular layer of the mature meander tail anterior lobe. (The ostensibly 'normal' meander tail posterior lobe also benefited from repletion of a more subtle granule cell deficiency.) Donor-derived neurons were well-integrated within the neuropil, suggesting that these progenitors' developmental programs for granule cell differentiation were unperturbed. These observations permitted several conclusions. (1) That exogenous progenitors could survive transplantation into affected regions of neonatal meander tail cerebellum and differentiate into the deficient cell type suggested that the microenvironment was not inimical to granule cell development. Rather it suggested that mea's deleterious action is intrinsic to the external granular layer cell. (Any cell-extrinsic actions- albeit unlikely--had to be restricted to readily circumventable prenatal events.) This study, therefore, offers a paradigm for using progenitors to help determine the site of action of other mutant genes or to test hypotheses regarding the pathophysiology underlying other anomalies. (2) In the regions most deficient in neurons, a neuronal phenotype was pursued in preference to other potential cell types, suggesting a 'push' of undifferentiated, multipotent progenitors towards compensation for granule cell dearth. These data suggested that progenitors with the potential for multiple fates might differentiate towards repletion of deficient cell types, a possible developmental mechanism with therapeutic implications. Neural progenitors (donor or endogenous) might enable cell replacement in some developmental or degenerative diseases--most obviously in cases where a defect is intrinsic to the diseased cell, but also, under certain circumstances, when extrinsic pathologic forces may exist. PMID- 9334272 TI - Epidermal induction and inhibition of neural fate by translation initiation factor 4AIII. AB - Bone Morphogenetic Protein-4 (BMP-4) is a potent epidermal inducer and inhibitor of neural fate. We have used differential screening to identify genes involved in epidermal induction downstream of BMP-4 and report here evidence of a novel translational mechanism that regulates the division of the vertebrate ectoderm into regions of neural and epidermal fate. In dissociated Xenopus ectoderm, addition of ectopic BMP-4 leads to an increase in the expression of translation initiation factor 4AIII (eIF-4AIII), a divergent member of the eIF-4A gene family until now characterized only in plants. In the gastrula embryo, Xenopus eIF-4AIII (XeIF-4AIII) expression is elevated in the ventral ectoderm, a site of active BMP signal transduction. Moreover, overexpression of XeIF-4AIII induces epidermis in dissociated cells that would otherwise adopt a neural fate, mimicking the effects of BMP-4. Epidermal induction by XeIF-4AIII requires both an active BMP signaling pathway and an extracellular intermediate. Our results suggest that XeIF-4AIII can regulate changes in cell fate through selective mRNA translation. We propose that BMPs and XeIF-4AIII interact through a positive feedback loop in the ventral ectoderm of the vertebrate gastrula. PMID- 9334273 TI - Notochord to endoderm signaling is required for pancreas development. AB - The role of the notochord in inducing and patterning adjacent neural and mesodermal tissues is well established. We provide evidence that the notochord is also required for one of the earliest known steps in the development of the pancreas, an endodermally derived organ. At a developmental stage in chick embryos when the notochord touches the endoderm, removal of notochord eliminates subsequent expression of several markers of dorsal pancreas bud development, including insulin, glucagon and carboxypeptidase A. Pancreatic gene expression can be initiated and maintained in prepancreatic chick endoderm grown in vitro with notochord. Non-pancreatic endoderm, however, does not express pancreatic genes when recombined with the same notochord. The results suggest that the notochord provides a permissive signal to endoderm to specify pancreatic fate in a stepwise manner. PMID- 9334275 TI - Suppressor of Hairless-independent events in Notch signaling imply novel pathway elements. AB - The Notch (N) pathway defines an evolutionarily conserved cell signaling mechanism that governs cell fate choices through local cell interactions. The ankyrin repeat region of the Notch receptor is essential for signaling and has been implicated in the interactions between Notch and two intracellular elements of the pathway: Deltex (Dx) and Suppressor of Hairless (Su(H)). Here we examine directly the function of the Notch cdc10/ankyrin repeats (ANK repeats) by transgenic and biochemical analysis. We present evidence implicating the ANK repeats in the regulation of Notch signaling through homotypic interactions. In vivo expression of the Notch ANK repeats reveals a cell non-autonomous effect and elicits mutant phenotypes that indicate the existence of novel downstream events in Notch signaling. These signaling activities are independent of the known effector Su(H) and suggest the existence of yet unidentified Notch pathway components. PMID- 9334274 TI - A role for FGF-8 in the dorsoventral patterning of the zebrafish gastrula. AB - Signals released from Spemann's organizer, together with ventralizing factors such as BMPs, are necessary to pattern the dorsoventral axis of the vertebrate embryo. We report that a member of the FGF family, fgf-8, not secreted by the axial mesoderm but expressed in a dorsoventral gradient at the margin of the zebrafish gastrula, also contributes to the establishment of the dorsoventral axis of the embryo. Ectopic expression of FGF-8 leads to the expansion of dorsolateral derivatives at the expense of ventral and posterior domains. Moreover, FGF-8 displays some organizer properties as it induces the formation of a partial secondary axis in the absence of factors released from Spemann's organizer territory. Analysis of its interaction with the ventralizing factors, BMPs, reveals that overexpression of FGF-8 inhibits the expression of these factors in the ventral part of the embryo as early as blastula stage, suggesting that FGF-8 acts upstream of BMP2 and BMP4. We conclude that FGF-8 is involved in defining dorsoventral identity and is an important organizing factor responsible for specification of mesodermal and ectodermal dorsolateral territories of the zebrafish gastrula. PMID- 9334276 TI - Animal and vegetal pole cells of early Xenopus embryos respond differently to maternal dorsal determinants: implications for the patterning of the organiser. AB - The maternal dorsal determinants required for the specification of the dorsal territories of Xenopus early gastrulae are located at the vegetal pole of unfertilised eggs and are moved towards the prospective dorsal region of the fertilised egg during cortical rotation. While the molecular identity of the determinants is unknown, there are dorsal factors in the vegetal cortical cytoplasm (VCC). Here, we show that the VCC factors, when injected into animal cells activate the zygotic genes Siamois and Xnr3, suggesting that they act along the Wnt/beta-catenin pathway. In addition, Siamois and Xnr3 are activated at the vegetal pole of UV-irradiated embryos, indicating that these two genes are targets of the VCC factors in all embryonic cells. However, the consequences of their activation in cells that occupy different positions along the animal vegetal axis differ. Dorsal vegetal cells of normal embryos or VCC-treated injected animal cells are able to dorsalise ventral mesoderm in conjugate experiments but UV-treated vegetal caps do not have this property. This difference is unlikely to reflect different levels of activation of FGF or activin-like signal transduction pathways but may reflect the activation of different targets of Siamois. Chordin, a marker of the head and axial mesoderm, is activated by the VCC/Siamois pathway in animal cells but not in vegetal cells whereas cerberus, a marker of the anterior mesendoderm which lacks dorsalising activity, can only be activated by the VCC/Siamois pathway in vegetal cells. We propose that the regionalisation of the organiser during gastrulation proceeds from the differential interpretation along the animal-vegetal axis of the activation of the VCC/beta-catenin/Siamois pathway. PMID- 9334277 TI - Neural tube-ectoderm interactions are required for trigeminal placode formation. AB - Cranial sensory ganglia in vertebrates develop from the ectodermal placodes, the neural crest, or both. Although much is known about the neural crest contribution to cranial ganglia, relatively little is known about how placode cells form, invaginate and migrate to their targets. Here, we identify Pax-3 as a molecular marker for placode cells that contribute to the ophthalmic branch of the trigeminal ganglion and use it, in conjunction with DiI labeling of the surface ectoderm, to analyze some of the mechanisms underlying placode development. Pax-3 expression in the ophthalmic placode is observed as early as the 4-somite stage in a narrow band of ectoderm contiguous to the midbrain neural folds. Its expression broadens to a patch of ectoderm adjacent to the midbrain and the rostral hindbrain at the 8- to 10-somite stage. Invagination of the first Pax-3 positive cells begins at the 13-somite stage. Placodal invagination continues through the 35-somite stage, by which time condensation of the trigeminal ganglion has begun. To challenge the normal tissue interactions leading to placode formation, we ablated the cranial neural crest cells or implanted barriers between the neural tube and the ectoderm. Our results demonstrate that, although the presence of neural crest cells is not mandatory for Pax-3 expression in the forming placode, a diffusible signal from the neuroectoderm is required for induction and/or maintenance of the ophthalmic placode. PMID- 9334278 TI - Complex regulatory region mediating tailless expression in early embryonic patterning and brain development. AB - tailless encodes a transcription factor expressed in multiple domains in the developing embryo. Early and transient expression at the posterior pole is required to establish a domain from which the eighth abdominal segment, telson and posterior gut arise. Just a few nuclear cycles later, a brain-specific domain is initiated at the anterior; expression in this domain is maintained with complex modulations throughout embryogenesis. Expression of tailless in this domain is required to establish the most anterior region of the brain. To understand the function and regulation of these different domains of expression, we provide a detailed description of tailless expression in brain neuroblasts and show that this expression is not detectably regulated by the head gap genes buttonhead or orthodenticle, by the proneural gene lethal of scute or by tailless itself. We show that approximately 6 kb of sequenced upstream regulatory DNA can drive lacZ expression in a pattern that mimics the full tailless embryonic expression pattern. Within this sequence we identify multiple modules responsible for different aspects of the tailless pattern. In addition to identifying additional torso response elements that mediate early blastoderm polar expression, we show that the complex brain expression pattern is driven by a combination of modules; thus expression at a low level throughout the brain and at a high level in the dorsal medial portion of the brain and in the optic lobe, as well as neuroblast-specific repression are mediated by different DNA regions. PMID- 9334279 TI - Mesodermal defects and cranial neural crest apoptosis in alpha5 integrin-null embryos. AB - Alpha5beta1 integrin is a cell surface receptor that mediates cell-extracellular matrix adhesions by interacting with fibronectin. Alpha5 subunit-deficient mice die early in gestation and display mesodermal defects; most notably, embryos have a truncated posterior and fail to produce posterior somites. In this study, we report on the in vivo effects of the alpha5-null mutation on cell proliferation and survival, and on mesodermal development. We found no significant differences in the numbers of apoptotic cells or in cell proliferation in the mesoderm of alpha5-null embryos compared to wild-type controls. These results suggest that changes in overall cell death or cell proliferation rates are unlikely to be responsible for the mesodermal deficits seen in the alpha5-null embryos. No increases in cell death were seen in alpha5-null embryonic yolk sac, amnion and allantois compared with wild-type, indicating that the mutant phenotype is not due to changes in apoptosis rates in these extraembryonic tissues. Increased numbers of dying cells were, however, seen in migrating cranial neural crest cells of the hyoid arch and in endodermal cells surrounding the omphalomesenteric artery in alpha5-null embryos, indicating that these subpopulations of cells are dependent on alpha5 integrin function for their survival. Mesodermal markers mox 1, Notch-1, Brachyury (T) and Sonic hedgehog (Shh) were expressed in the mutant embryos in a regionally appropriate fashion. Both T and Shh, however, showed discontinuous expression in the notochords of alpha5-null embryos due to (1) degeneration of the notochordal tissue structure, and (2) non-maintenance of gene expression. Consistent with the disorganization of notochordal signals in the alpha5-null embryos, reduced Pax-1 expression and misexpression of Pax-3 were observed. Anteriorly expressed HoxB genes were expressed normally in the alpha5 null embryos. However, expression of the posteriormost HoxB gene, Hoxb-9, was reduced in alpha5-null embryos. These results suggest that alpha5beta1 fibronectin interactions are not essential for the initial commitment of mesodermal cells, but are crucial for maintenance of mesodermal derivatives during postgastrulation stages and also for the survival of some neural crest cells. PMID- 9334281 TI - The terminal differentiation factor LIN-29 is required for proper vulval morphogenesis and egg laying in Caenorhabditis elegans. AB - Caenorhabditis elegans vulval development culminates during exit from the L4-to adult molt with the formation of an opening through the adult hypodermis and cuticle that is used for egg laying and mating. Vulva formation requires the heterochronic gene lin-29, which triggers hypodermal cell terminal differentiation during the final molt. lin-29 mutants are unable to lay eggs or mate because no vulval opening forms; instead, a protrusion forms at the site of the vulva. We demonstrate through analysis of genetic mosaics that lin-29 is absolutely required in a small subset of lateral hypodermal seam cells, adjacent to the vulva, for wild-type vulva formation and egg laying. However, lin-29 function is not strictly limited to the lateral hypodermis. First, LIN-29 accumulates in many non-hypodermal cells with known roles in vulva formation or egg laying. Second, animals homozygous for one lin-29 allele, ga94, have the vulval defect and cannot lay eggs, despite having a terminally differentiated adult lateral hypodermis. Finally, vulval morphogenesis and egg laying requires lin-29 activity within the EMS lineage, a lineage that does not generate hypodermal cells. PMID- 9334280 TI - muscleblind, a gene required for photoreceptor differentiation in Drosophila, encodes novel nuclear Cys3His-type zinc-finger-containing proteins. AB - We have isolated the embryonic lethal gene muscleblind (mbl) as a suppressor of the sev-svp2 eye phenotype. Analysis of clones mutant for mbl during eye development shows that it is autonomously required for photoreceptor differentiation. Mutant cells are recruited into developing ommatidia and initiate neural differentiation, but they fail to properly differentiate as photoreceptors. Molecular analysis reveals that the mbl locus is large and complex, giving rise to multiple different proteins with common 5' sequences but different carboxy termini. Mbl proteins are nuclear and share a Cys3His zinc finger motif which is also found in the TIS11/NUP475/TTP family of proteins and is highly conserved in vertebrates and invertebrates. Functional analysis of mbl, the observation that it also dominantly suppresses the sE-Jun(Asp) gain-of function phenotype and the phenotypic similarity to mutants in the photoreceptor specific glass gene suggest that mbl is a general factor required for photoreceptor differentiation. PMID- 9334282 TI - A silencer is required for maintenance of transcriptional repression throughout Drosophila development. AB - Transcriptional silencing by the Polycomb Group of genes maintains the position specific repression of homeotic genes throughout Drosophila development. The Polycomb Group of genes characterized to date encode chromatin-associated proteins that have been suggested to form heterochromatin-like structures. By studying the expression of reporter genes, we have identified a 725 bp fragment, called MCP725, in the homeotic gene Abdominal-B, that accurately maintains position-specific silencing during proliferation of imaginal cells. Silencing by MCP725 requires the Polycomb and the Polycomblike genes, indicating that it contains a Polycomb response element To investigate the mechanisms of transcriptional silencing by MCP725, we have studied its temporal requirements by removing MCP725 from the transgene at various times during development. We have discovered that excision of MCP725 during larval stages leads to loss of silencing. Our findings indicate that the silencer is required for the maintenance of the repressed state throughout cell proliferation. They also suggest that propagation of the silenced state does not occur merely by templating of a heterochromatin structure by virtue of protein-protein interactions. Rather, they suggest that silencers play an active role in the maintenance of the position-specific repression throughout development. PMID- 9334283 TI - Timing and pattern of cell fate restrictions in the neural crest lineage. AB - The trunk neural crest of vertebrate embryos is a transient collection of precursor cells present along the dorsal aspect of the neural tube. These cells migrate on two distinct pathways and give rise to specific derivatives in precise embryonic locations. One group of crest cells migrates early on a ventral pathway and generates neurons and glial cells. A later-dispersing group migrates laterally and gives rise to melanocytes in the skin. These observations raise the possibility that the appearance of distinct derivatives in different embryonic locations is a consequence of lineage restrictions specified before or soon after the onset of neural crest cell migration. To test this notion, we have assessed when and in what order distinct cell fates are specified during neural crest development. We determined the proportions of different types of precursor cells in cultured neural crest populations immediately after emergence from the neural tube and at intervals as development proceeds. We found that the initial neural crest population was a heterogeneous mixture of precursors almost half of which generated single-phenotype clones. Distinct neurogenic and melanogenic sublineages were also present in the outgrowth population almost immediately, but melanogenic precursors dispersed from the neural tube only after many neurogenic precursors had already done so. A discrete fate-restricted neuronal precursor population was distinguished before entirely separate fate-restricted melanocyte and glial precursor populations were present, and well before initial neuronal differentiation. Taken together, our results demonstrate that lineage-restricted subpopulations constitute a major portion of the initial neural crest population and that neural crest diversification occurs well before overt differentiation by the asynchronous restriction of distinct cell fates. Thus, the different morphogenetic and differentiative behavior of neural crest subsets in vivo may result from earlier cell fate-specification events that generate developmentally distinct subpopulations that respond differentially to environmental cues. PMID- 9334284 TI - bag-of-marbles and benign gonial cell neoplasm act in the germline to restrict proliferation during Drosophila spermatogenesis. AB - Stem cells divide asymmetrically, regenerating a parental stem cell and giving rise to a daughter cell with a distinct fate. In many stem cell lineages, this daughter cell undergoes several amplificatory mitoses, thus generating more cells that embark on the differentiation program specific for the given lineage. Spermatogenesis in Drosophila is a model system to identify molecules regulating stem cell lineages. Mutations at two previously identified loci, bag-of-marbles (bam) and benign gonial cell neoplasm (bgcn), prevent progression through spermatogenesis and oogenesis, resulting in the overproliferation of undifferentiated germ cells. Here we investigate how bam and bgcn regulate the male germline stem cell lineage. By generating FLP-mediated clones, we demonstrate that both bam and bgcn act autonomously in the germline to restrict proliferation during spermatogenesis. By using enhancer trap lines, we find that the overproliferating germ cells express markers specific to amplifying germ cells, while at the same time retaining the expression of some markers of stem cell and primary spermatogonial cell fate. However, we find that germ cells accumulating in bam or bgcn mutant testes most resemble amplifying germ cells, because they undergo incomplete cytokinesis and progress through the cell cycle in synchrony within a cyst, which are two characteristics of amplifying germ cells, but not of stem cells. Taken together, our results suggest that bam and bgcn regulate progression through the male germline stem cell lineage by cell intrinsically restricting the proliferation of amplifying germ cells. PMID- 9334285 TI - Left-right pattern of cardiac BMP4 may drive asymmetry of the heart in zebrafish. AB - The first evident break in left-right symmetry of the primitive zebrafish heart tube is the shift in pattern of BMP4 expression from radially symmetric to left predominant. The midline heart tube then 'jogs' to the left and subsequently loops to the right. We examined 279 mutations, affecting more than 200 genes, and found 21 mutations that perturb this process. Some cause BMP4 to remain radially symmetric. Others randomize the asymmetric BMP4 pattern. Retention of BMP4 symmetry is associated with failure to jog: right-predominance of the BMP4 pattern is associated with reversal of the direction of jogging and looping. Raising BMP4 diffusely throughout the heart, via sonic hedgehog injection, or the blocking of its action by injection of a dominant negative BMP4 receptor, prevent directional jogging or looping. The genes crucial to directing cardiac asymmetry include a subset of those needed for patterning the dorsoventral axis and for notochord and ventral spinal cord development. Thus, the pattern of cardiac BMP4 appears to be in the pathway by which the heart interprets lateralizing signals from the midline. PMID- 9334286 TI - punt and schnurri regulate a somatically derived signal that restricts proliferation of committed progenitors in the germline. AB - To identify regulators of stem cell lineages, we are focusing on spermatogenesis in Drosophila. In spermatogenesis, each germline stem cell divides asymmetrically, renewing itself and producing a transiently amplifying daughter, which divides four times. By screening for mutants in which daughter cells fail to stop dividing, we find that the TGF-beta signal transducers schnurri and punt are required to limit transient amplification of germ cells. Mosaic analysis demonstrates that punt and schnurri act within somatic cyst cells that surround germ cells, rather than in germ cells. Thus, a cyst-cell-derived signal restricts germ cell proliferation and this signal is initiated by input from a member of the TGF-beta superfamily. Thus, a signal relay regulates progression through the germline stem cell lineage. PMID- 9334287 TI - Relationship between dose, distance and time in Sonic Hedgehog-mediated regulation of anteroposterior polarity in the chick limb. AB - Anteroposterior polarity in the vertebrate limb is thought to be regulated in response to signals derived from a specialized region of distal posterior mesenchyme, the zone of polarizing activity. Sonic Hedgehog (Shh) is expressed in the zone of polarizing activity and appears to mediate the action of the zone of polarizing activity. Here we have manipulated Shh signal in the limb to assess whether it acts as a long-range signal to directly pattern all the digits. Firstly, we demonstrate that alterations in digit development are dependent upon the dose of Shh applied. DiI-labeling experiments indicate that cells giving rise to the extra digits lie within a 300 microm radius of a Shh bead and that the most posterior digits come from cells that lie very close to the bead. A response to Shh involves a 12-16 hour period in which no irreversible changes in digit pattern occur. Increasing the time of exposure to Shh leads to specification of additional digits, firstly digit 2, then 3, then 4. Cell marking experiments demonstrate that cells giving rise to posterior digits are first specified as anterior digits and later adopt a more posterior character. To monitor the direct range of Shh signalling, we developed sensitive assays for localizing Shh by attaching alkaline phosphatase to Shh and introducing cells expressing these forms into the limb bud. These experiments demonstrate that long-range diffusion across the anteroposterior axis of the limb is possible. However, despite a dramatic difference in their diffusibility in the limb mesenchyme, the two forms of alkaline phosphatase-tagged Shh proteins share similar polarizing activity. Moreover, Shh-N (aminoterminal peptide of Shh)-coated beads and Shh-expressing cells also exhibit similar patterning activity despite a significant difference in the diffusibility of Shh from these two sources. Finally, we demonstrate that when Shh-N is attached to an integral membrane protein, cells transfected with this anchored signal also induce mirror-image pattern duplications in a dose dependent fashion similar to the zone of polarizing activity itself. These data suggest that it is unlikely that Shh itself signals digit formation at a distance. Beads soaked in Shh-N do not induce Shh in anterior limb mesenchyme ruling out direct propagation of a Shh signal. However, Shh induces dose dependent expression of Bmp genes in anterior mesenchyme at the start of the promotion phase. Taken together, these results argue that the dose-dependent effects of Shh in the regulation of anteroposterior pattern in the limb may be mediated by some other signal(s). BMPs are plausible candidates. PMID- 9334289 TI - Mutations in the HYDRA1 gene of Arabidopsis perturb cell shape and disrupt embryonic and seedling morphogenesis. AB - Mutations in the HYDRA1 (HYD1) gene of Arabidopsis thaliana can prevent normal morphological development of embryos and seedlings. Three allelic mutants (hydra 1-1, hydra1-2 and hydra1-3) have been identified, and in each the seedling is characterized by having a variable number of cotyledons, a short and wide hypocotyl and a much reduced root system. hydra1 embryos appear to develop normally to the octant stage, but fail to establish a distinct protoderm and lack bilateral symmetry, developing multiple cotyledonary primordia of irregular size and shape. Cells of the embryo proper, but not the suspensor, exhibit abnormalities in size and shape. The hydra1 embryo fails to develop an embryonic root, but embryos and seedlings express molecular markers of apical-basal polarity. Mutant seedlings produce leaves to form a small cabbage-like habit and may occasionally produce sterile flowers, though the mutation is commonly seedling-lethal. hydra1 seedlings exhibit abnormal radial patterning, but nevertheless express at least one molecular marker of vascular cell differentiation. A model is proposed in which the HYDRA1 protein functions as an essential component of the cell expansion system. PMID- 9334288 TI - Forced expression of the homeodomain protein Gax inhibits cardiomyocyte proliferation and perturbs heart morphogenesis. AB - The development of the tubular heart into a complex four-chambered organ requires precise temporal and region-specific regulation of cell proliferation, migration, death and differentiation. While the regulatory mechanisms in heart morphogenesis are not well understood, increasing attention has focused on the homeodomain proteins, which are generally linked to morphogenetic processes. The homeodomain containing gene Gax has been shown to be expressed in heart and smooth muscle tissues. In this study, the Gax protein was detected in the nuclei of myocardial cells relatively late in chicken heart development, at a time when myocyte proliferation is declining. To test the hypothesis that the Gax protein functions as a negative regulator of cardiomyocyte proliferation, a replication-defective adenovirus was used to force its precocious nuclear expression during chicken heart morphogenesis. In experiments in which Gax- and beta-galactosidase expressing adenoviruses were co-injected, clonal expansion of myocytes was reduced, consistent with inhibition of myocyte proliferation. This effect on proliferation was corroborated by the finding that the percentage of exogenous Gax-expressing myocytes that were positive for the cell cycle marker PCNA decreased over time and was lower than in control myocytes. The precocious nuclear expression of Gax in tubular hearts resulted in abnormal heart morphology, including small ventricles with rounded apices, a thinned compact zone and coarse trabeculae. These results suggest a role for the Gax protein in heart morphogenesis causing proliferating cardiomyocytes to withdraw from the cell cycle, thus influencing the size and shape that the heart ultimately attains. PMID- 9334290 TI - Role of MMTV integration locus cellular genes in breast cancer. AB - Mouse mammary tumorigenesis as a result of mouse mammary tumor virus (MMTV) integrations has helped to identify a wide variety of interesting genes that play a role in mammary development and tumorigenesis. Several such genes int1/wnt1, wnt3, wnt 10B, int2/fgf3, fgf4, int3/notch and int6 have been shown to be genetically altered in naturally formed mammary tumors as a consequence of MMTV integration. Some of these genes have been well characterised and examined in in vivo breast cancer transgenic models for their potential for tumorigenesis. Overexpression of one or more of these genes have resulted in a striking proliferation of mammary gland epithelium of both female and male transgenic mice. Our own studies have demonstrated overexpression of int5/aromatase in mammary glands of virgin and postlactational females leads to the induction of various preneoplastic and neoplastic changes that are similar to early breast cancer, that may, in turn, increase the risks for developing breast cancer. Therefore, further understanding of these genes should provide new insights to their involvement and mechanism of action in breast cancer. PMID- 9334291 TI - Genetics of aging. AB - The role of genetics in determining life-span is complex and paradoxical. Although the heritability of life-span is relatively minor, some genetic variants significantly modify senescence of mammals and invertebrates, with both positive and negative impacts on age-related disorders and life-spans. In certain examples, the gene variants alter metabolic pathways, which could thereby mediate interactions with nutritional and other environmental factors that influence life span. Given the relatively minor effect and variable penetrance of genetic risk factors that appear to affect survival and health at advanced ages, life-style and other environmental influences may profoundly modify outcomes of aging. PMID- 9334292 TI - Life and death of neurons in the aging brain. AB - Neurodegenerative disorders are characterized by extensive neuron death that leads to functional decline, but the neurobiological correlates of functional decline in normal aging are less well defined. For decades, it has been a commonly held notion that widespread neuron death in the neocortex and hippocampus is an inevitable concomitant of brain aging, but recent quantitative studies suggest that neuron death is restricted in normal aging and unlikely to account for age-related impairment of neocortical and hippocampal functions. In this article, the qualitative and quantitative differences between aging and Alzheimer's disease with respect to neuron loss are discussed, and age-related changes in functional and biochemical attributes of hippocampal circuits that might mediate functional decline in the absence of neuron death are explored. When these data are viewed comprehensively, it appears that the primary neurobiological substrates for functional impairment in aging differ in important ways from those in neurodegenerative disorders such as Alzheimer's disease. PMID- 9334293 TI - The endocrinology of aging. AB - Most aging individuals die from atherosclerosis, cancer, or dementia; but in the oldest old, loss of muscle strength resulting in frailty is the limiting factor for an individual's chances of living an independent life until death. Three hormonal systems show decreasing circulating hormone concentrations during normal aging: (i) estrogen (in menopause) and testosterone (in andropause), (ii) dehydroepiandrosterone and its sulphate (in adrenopause), and (iii) the growth hormone/insulin-like growth factor I axis (in somatopause). Physical changes during aging have been considered physiologic, but there is evidence that some of these changes are related to this decline in hormonal activity. Hormone replacement strategies have been developed, but many of their aspects remain controversial, and increasing blood hormone levels in aging individuals to those found during mid-adult life has not been uniformly proven to be safe and of benefit. PMID- 9334294 TI - The structure of nitric oxide synthase oxygenase domain and inhibitor complexes. AB - The nitric oxide synthase oxygenase domain (NOSox) oxidizes arginine to synthesize the cellular signal and defensive cytotoxin nitric oxide (NO). Crystal structures determined for cytokine-inducible NOSox reveal an unusual fold and heme environment for stabilization of activated oxygen intermediates key for catalysis. A winged beta sheet engenders a curved alpha-beta domain resembling a baseball catcher's mitt with heme clasped in the palm. The location of exposed hydrophobic residues and the results of mutational analysis place the dimer interface adjacent to the heme-binding pocket. Juxtaposed hydrophobic O2- and polar L-arginine-binding sites occupied by imidazole and aminoguanidine, respectively, provide a template for designing dual-function inhibitors and imply substrate-assisted catalysis. PMID- 9334297 TI - Triton's distorted atmosphere. AB - A stellar-occultation light curve for Triton shows asymmetry that can be understood if Triton's middle atmosphere is distorted from spherical symmetry. Although a globally oblate model can explain the data, the inferred atmospheric flattening is so large that it could be caused only by an unrealistic internal mass distribution or highly supersonic zonal winds. Cyclostrophic winds confined to a jet near Triton's northern or southern limbs (or both) could also be responsible for the details of the light curve, but such winds are required to be slightly supersonic. Hazes and clouds in the atmosphere are unlikely to have caused the asymmetry in the light curve. PMID- 9334299 TI - Direct observation of cooling of heme upon photodissociation of carbonmonoxy myoglobin. AB - The formation of vibrationally excited heme upon photodissociation of carbonmonoxy myoglobin and its subsequent vibrational energy relaxation was monitored by picosecond anti-Stokes resonance Raman spectroscopy. The anti-Stokes intensity of the nu4 band showed immediate generation of vibrationally excited hemes and biphasic decay of the excited populations. The best fit to double exponentials gave time constants of 1.9 +/- 0.6 and 16 +/- 9 picoseconds for vibrational population decay and 3.0 +/- 1.0 and 25 +/- 14 picoseconds for temperature relaxation of the photolyzed heme when a Boltzmann distribution was assumed. The decay of the nu4 anti-Stokes intensity was accompanied by narrowing and frequency upshift of the Stokes counterpart. This direct monitoring of the cooling dynamics of the heme cofactor within the globin matrix allows the characterization of the vibrational energy flow through the protein moiety and to the water bath. PMID- 9334300 TI - DNA solution of the maximal clique problem. AB - The maximal clique problem has been solved by means of molecular biology techniques. A pool of DNA molecules corresponding to the total ensemble of six vertex cliques was built, followed by a series of selection processes. The algorithm is highly parallel and has satisfactory fidelity. This work represents further evidence for the ability of DNA computing to solve NP-complete search problems. PMID- 9334302 TI - The homeotic gene lin-39 and the evolution of nematode epidermal cell fates. AB - The fate of ventral epidermal cells differs among nematode species. Nonvulval cells fuse with the epidermis in Caenorhabditis elegans, whereas the homologous cells undergo apoptosis in Pristionchus pacificus. The homeotic gene lin-39 is involved in the regulation of these epidermal cell fates. In Caenorhabditis, lin 39 prevents cell fusion of potential vulval cells and specifies the vulva equivalence group. Pristionchus vulvaless mutants that displayed apoptosis of the vulval precursor cells were isolated, and point mutations in lin-39 were identified. Thus, the evolution of these epidermal cell fates is driven by different intrinsic properties of homologous cells. PMID- 9334303 TI - Phosphorylation of Sic1p by G1 Cdk required for its degradation and entry into S phase. AB - G1 cyclin-dependent kinase (Cdk)-triggered degradation of the S-phase Cdk inhibitor Sic1p has been implicated in the transition from G1 to S phase in the cell cycle of budding yeast. A multidimensional electrospray mass spectrometry technique was used to map G1 Cdk phosphorylation sites in Sic1p both in vitro and in vivo. A Sic1p mutant lacking three Cdk phosphorylation sites did not serve as a substrate for Cdc34p-dependent ubiquitination in vitro, was stable in vivo, and blocked DNA replication. Moreover, purified phosphoSic1p was ubiquitinated in cyclin-depleted G1 extract, indicating that a primary function of G1 cyclins is to tag Sic1p for destruction. These data suggest a molecular model of how phosphorylation and proteolysis cooperate to bring about the G1/S transition in budding yeast. PMID- 9334304 TI - CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. AB - Proteolysis mediated by the anaphase-promoting complex (APC) triggers chromosome segregation and exit from mitosis, yet its regulation is poorly understood. The conserved Cdc20 and Cdh1 proteins were identified as limiting, substrate-specific activators of APC-dependent proteolysis. CDC20 was required for the degradation of the APC substrate Pds1 but not for that of other APC substrates, such as Clb2 and Ase1. Conversely, cdh1Delta mutants were impaired in the degradation of Ase1 and Clb2 but not in that of Pds1. Overexpression of either CDC20 or CDH1 was sufficient to induce APC-dependent proteolysis of the appropriate target in stages of the cell cycle in which substrates are normally stable. PMID- 9334305 TI - Forward and backward propagation of dendritic impulses and their synaptic control in mitral cells. AB - The site of impulse initiation is crucial for the integrative actions of mammalian central neurons, but this question is currently controversial. Some recent studies support classical evidence that the impulse always arises in the soma-axon hillock region, with back-propagation through excitable dendrites, whereas others indicate that the dendrites are sufficiently excitable to initiate impulses that propagate forward along the dendrite to the soma-axon hillock. This issue has been addressed in the olfactory mitral cell, in which excitatory synaptic input is restricted to the distal tuft of a single primary dendrite. In rat olfactory bulb slices, dual whole cell recordings were made at or near the soma and from distal sites on the primary dendrite. The results show that the impulse can be initiated in either the soma-axon hillock or in the distal primary dendrite, and that the initiation site is controlled physiologically by the excitatory synaptic inputs to the distal tuft and inhibitory synaptic inputs near the soma. PMID- 9334306 TI - Mediation of classical conditioning in Aplysia californica by long-term potentiation of sensorimotor synapses. AB - Long-term potentiation (LTP) is considered an important neuronal mechanism of learning and memory. Currently, however, there is no direct experimental link between LTP of an identified synapse and learning. A cellular analog of classical conditioning in Aplysia was used to determine whether this form of invertebrate learning involves N-methyl-D-aspartate (NMDA)-type LTP. The NMDA receptor antagonist dl-2-amino-5-phosphonovalerate significantly disrupted synaptic enhancement after associative training but did not disrupt synaptic enhancement after nonassociative training. Thus, classical conditioning in Aplysia appears to be mediated, in part, by LTP due to activation of NMDA-related receptors. PMID- 9334307 TI - Inhibition of phosphatases and increased Ca2+ channel activity by inositol hexakisphosphate. AB - Inositol hexakisphosphate (InsP6), the dominant inositol phosphate in insulin secreting pancreatic beta cells, inhibited the serine-threonine protein phosphatases type 1, type 2A, and type 3 in a concentration-dependent manner. The activity of voltage-gated L-type calcium channels is increased in cells treated with inhibitors of serine-threonine protein phosphatases. Thus, the increased calcium channel activity obtained in the presence of InsP6 might result from the inhibition of phosphatase activity. Glucose elicited a transient increase in InsP6 concentration, which indicates that this inositol polyphosphate may modulate calcium influx over the plasma membrane and serve as a signal in the pancreatic beta cell stimulus-secretion coupling. PMID- 9334308 TI - Interneuron migration from basal forebrain to neocortex: dependence on Dlx genes. AB - Although previous analyses indicate that neocortical neurons originate from the cortical proliferative zone, evidence suggests that a subpopulation of neocortical interneurons originates within the subcortical telencephalon. For example, gamma-aminobutyric acid (GABA)-expressing cells migrate in vitro from the subcortical telencephalon into the neocortex. The number of GABA-expressing cells in neocortical slices is reduced by separating the neocortex from the subcortical telencephalon. Finally, mice lacking the homeodomain proteins DLX-1 and DLX-2 show no detectable cell migration from the subcortical telencephalon to the neocortex and also have few GABA-expressing cells in the neocortex. PMID- 9334309 TI - Regulation of gliogenesis in the central nervous system by the JAK-STAT signaling pathway. AB - A mechanism by which members of the ciliary neurotrophic factor (CNTF)-leukemia inhibitory factor cytokine family regulate gliogenesis in the developing mammalian central nervous system was characterized. Activation of the CNTF receptor promoted differentiation of cerebral cortical precursor cells into astrocytes and inhibited differentiation of cortical precursors along a neuronal lineage. Although CNTF stimulated both the Janus kinase-signal transducer and activator of transcription (JAK-STAT) and Ras-mitogen-activated protein kinase signaling pathways in cortical precursor cells, the JAK-STAT signaling pathway selectively enhanced differentiation of these precursors along a glial lineage. These findings suggest that cytokine activation of the JAK-STAT signaling pathway may be a mechanism by which cell fate is controlled during mammalian development. PMID- 9334310 TI - Peripheral and cerebral asymmetries in the rat. AB - Rats learn a novel foraging pattern better with their right-side whiskers than with their left-side whiskers. They also learn better with the left cerebral hemisphere than with the right hemisphere. Rotating an already learned maze relative to the external environment most strongly reduces right-whisker performance; starting an already learned maze at a different location most strongly reduces left-whisker performance. These results suggest that the right periphery-left-hemisphere system accesses a map-like representation of the foraging problem, whereas the left-periphery-right-hemisphere system accesses a rote path. Thus, as in humans, functional asymmetries in rats can be elicited by both peripheral and cortical manipulation, and each hemisphere makes qualitatively distinct contributions to a complex natural behavior. PMID- 9334311 TI - Link between aging and the nucleolus. PMID- 9334312 TI - The PAG gene product, a stress-induced protein with antioxidant properties, is an Abl SH3-binding protein and a physiological inhibitor of c-Abl tyrosine kinase activity. AB - Biochemical and genetic evidence suggests that the tyrosine kinase activity of c Abl is tightly regulated in vivo by a cellular factor binding to the Src homology 3 (SH3) domain of Abl. We used the yeast two-hybrid system to identify a gene, PAG, whose protein product (Pag) interacts specifically with the Abl SH3 domain. Pag, also known as macrophage 23-kD stress protein (MSP23), is a member of a novel family of proteins with antioxidant activity implicated in the cellular response to oxidative stress and in control of cell proliferation and differentiation. In a co-expression assay, Pag associates with c-Abl in vivo and inhibits tyrosine phosphorylation induced by overexpression of c-Abl. Inhibition requires the Abl SH3 and kinase domains and is not observed with other Abl SH3 binding proteins. Expression of Pag also inhibits the in vitro kinase activity of c-Abl, but not SH3-mutated Abl or v-Abl. When transfected in NIH-3T3 cells, Pag is localized to nucleus and cytoplasm and rescues the cytostatic effect induced by c-Abl. These observations suggest Pag is a physiological inhibitor of c-Abl in vivo. PMID- 9334313 TI - K-ras is an essential gene in the mouse with partial functional overlap with N ras. AB - Mammalian ras genes are thought to be critical in the regulation of cellular proliferation and differentiation and are mutated in approximately 30% of all human tumors. However, N-ras and H-ras are nonessential for mouse development. To characterize the normal role of K-ras in growth and development, we have mutated it by gene targeting in the mouse. On an inbred genetic background, embryos homozygous for this mutation die between 12 and 14 days of gestation, with fetal liver defects and evidence of anemia. Thus, K-ras is the only member of the ras gene family essential for mouse embryogenesis. We have also investigated the effect of multiple mutations within the ras gene family. Most animals lacking N ras function and heterozygous for the K-ras mutation exhibit abnormal hematopoietic development and die between days 10 and 12 of embryogenesis. Thus, partial functional overlap appears to occur within the ras gene family, but K-ras provides a unique and essential function. PMID- 9334314 TI - A multifunctional DNA-binding protein that promotes the formation of serum response factor/homeodomain complexes: identity to TFII-I. AB - The human homeodomain protein Phox1 interacts functionally with serum response factor (SRF) to impart serum responsive transcriptional activity to SRF-binding sites in a HeLa cell cotransfection assay. However, stable ternary complexes composed of SRF, Phox1, and DNA, which presumably mediate the transcriptional effects of Phox1 in vivo, have not been observed in vitro. Here, we report the identification, purification, and molecular cloning of a human protein that promotes the formation of stable higher-order complexes of SRF and Phox1. We show that this protein, termed SPIN, interacts with SRF and Phox1 in vitro and in vivo. Furthermore, SPIN binds specifically to multiple sequences in the c-fos promoter and interacts cooperatively with Phox1 to promote serum-inducible transcription of a reporter gene driven by the c-fos serum response element (SRE). SPIN is identical to the initiator-binding protein TFII-I. Consistent with this hypothesis, SPIN exhibits modest affinity for a characterized initiator sequence in vitro. We propose that this multifunctional protein coordinates the formation of an active promoter complex at the c-fos gene, including the linkage of specific signal responsive activator complexes to the general transcription machinery. PMID- 9334315 TI - Intergenic transcription and transinduction of the human beta-globin locus. AB - We have identified novel nuclear transcripts in the human beta-globin locus using nuclear run-on analysis in erythroid cell lines and in situ hybridization analysis of erythroid tissue. These transcripts extend across the LCR and intergenic regions but are undetectable in nonerythroid cells. Surprisingly, transient transfection of a beta-globin gene (epsilon, gamma, or beta) induces transcription of the LCR and intergenic regions from the chromosomal beta-globin locus in nonerythroid cell lines. The beta-globin genes themselves, however, remain transcriptionally silent. Induction is dependent on transcription of the globin gene in the transfected plasmid but does not require protein expression. Using in situ hybridization analysis, we show that the plasmid colocalizes with the endogenous beta-globin locus providing insight into the mechanism of transinduction. PMID- 9334316 TI - Translational repressor bruno plays multiple roles in development and is widely conserved. AB - oskar (osk) mRNA is tightly localized to the posterior pole of the Drosophila oocyte, where the subsequent expression of Osk protein directs abdomen and germ line formation in the developing embryo. Misplaced expression of Osk protein leads to lethal body patterning defects. The Osk message is translationally repressed before and during the localization process, ensuring that Osk protein is only expressed after the mRNA has reached the posterior. An ovarian protein, Bruno (Bru), has been implicated as a translational repressor of osk mRNA. Here we report the isolation of a cDNA encoding Bru using a novel approach to the expression cloning of an RNA-binding protein, and the identification of previously described mutants in the arrest (aret)-locus as mutants in Bru. The mutant phenotype, along with the binding properties of the protein and its pattern of accumulation within the oocyte, indicate that Bru regulates multiple mRNAs involved in female and male gametogenesis as well as early in embryogenesis. Genetic experiments provide further evidence that Bru functions in the translational repression of osk. Intriguingly, we find that Bru interacts physically with Vasa (Vas), an RNA helicase that is a positive regulator of osk translation. Bru belongs to an evolutionarily conserved family of genes, suggesting that Bru-mediated translational regulation may be widespread. Models for the molecular mechanism of Bru function are discussed. PMID- 9334317 TI - Coupling of cell division to cell growth by translational control of the G1 cyclin CLN3 in yeast. AB - The eukaryotic cell cycle is driven by a cascade of cyclins and kinase partners including the G1 cyclin Cln3p in yeast. As the first step in this cascade, Cln3p is uniquely positioned to determine the critical growth-rate threshold for division. To analyze factors regulating CLN3 expression, we identified a short upstream open reading frame (uORF) in the 5' leader of CLN3 mRNA as a translational control element. This control element is critical for the growth dependent regulation of Cln3p synthesis because it specifically represses CLN3 expression during conditions of diminished protein synthesis or slow growth. Inactivation of the uORF accelerates the completion of Start and entry into the cell cycle suggesting that translational regulation of CLN3 provides a mechanism coupling cell growth and division. PMID- 9334318 TI - Inhibition of patterned cell shape change and cell invasion by Discs large during Drosophila oogenesis. AB - Drosophila Discs large (Dlg) is a tumor suppressor gene whose loss in epithelial tissues causes disrupted cell polarity and increased cell proliferation. A human Dlg homolog, hDlg, has been implicated in tumorigenic processes via its association with the product of the Adenomatous Polyposis Coli (APC) gene. We show for the first time that Drosophila Dlg is required to block cell invasion. Loss of dlg activity during oogenesis causes follicle cells to change shape and invade in a pattern similar to border cells, a small population of cells that break from the post-mitotic follicular epithelium during wild-type oogenesis, yet dlg mutant cells have not adopted a border cell fate. Both functional and morphological evidence indicates that cooperation between germ cell and follicle cell Dlg, probably mediated by Dlg PDZ domains, is crucial for regulating cell mixing, suggesting a novel developmental mechanism and mode of action for the Dlg family of molecules. These findings suggest that Dlg does not simply inhibit individual cell behaviors during oogenesis, but rather acts in a developmental pathway essential for blocking cell proliferation and migration in a spatio temporally defined manner. A model for Dlg action in blocking cell invasion is presented. PMID- 9334319 TI - Hrp1, a sequence-specific RNA-binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3'-end formation in yeast. AB - In yeast, four factors (CF I, CF II, PF I, and PAP) are required for accurate pre mRNA cleavage and polyadenylation in vitro. CF I can be separated further into CF IA and CF IB. Here we show that CF IB is the 73-kD Hrp1 protein. Recombinant Hrp1p made in Escherichia coli provides full CF IB function in both cleavage and poly(A) addition assays. Consistent with the presence of two RRM-type motifs, Hrp1p can be UV cross-linked to RNA, and this specific interaction requires the (UA)6 polyadenylation efficiency element. Furthermore, the CF II factor enhances the binding of Hrp1p to the RNA precursor. A temperature-sensitive mutant in HRP1 yields mRNAs with shorter poly(A) tails when grown at the nonpermissive temperature. Genetic analyses indicate that Hrp1p interacts with Rna15p and Rna14p, two components of CF 1A. The HRP1 gene was originally isolated as a suppressor of a temperature-sensitive npl3 allele, a gene encoding a protein involved in mRNA export. Like Npl3p, Hrp1p shuttles between the nucleus and cytoplasm, providing a potential link between 3'-end processing and mRNA export from the nucleus. PMID- 9334320 TI - AU-rich elements target small nuclear RNAs as well as mRNAs for rapid degradation. AB - AU-rich elements (AREs, usually containing repeated copies of AUUUA), when present in the 3'-untranslated regions (UTRs) of many mammalian mRNAs, confer instability on their host RNA molecules. The viral small nuclear RNA (snRNA) Herpesvirus saimiri U RNA 1 (HSUR 1) also contains an AUUUA-rich sequence. Here, we report that this ARE induces rapid degradation of HSUR 1 itself and of other snRNAs including HSUR 2 and cellular U1. Mutational analyses of the viral ARE establish that sequence requirements for mRNA and snRNA decay are the same, suggesting a similar mechanism. Moreover, the in vivo degradation activity of mutant AREs correlates with their in vitro binding to the HuR protein, implicated previously as a component of the mRNA degradation machinery. Our results suggest that ARE-mediated instability can be uncoupled from both ongoing translation and deadenylation of the target RNA. PMID- 9334321 TI - A novel histidine kinase inhibitor regulating development in Bacillus subtilis. AB - Kinase A is the sensor histidine kinase responsible for processing postexponential phase information and providing phosphate input to the phosphorelay that activates developmental transcription via phosphorylated Spo0A. A protein inhibitor, KipI, of kinase A was discovered encoded in an operon of genes of unknown function but regulated by the availability of fixed nitrogen. KipI is a potent inhibitor of the autophosphorylation reaction of kinase A but does not inhibit phosphate transfer to the Spo0F response regulator once kinase A is phosphorylated. KipI is an inhibitor of the catalytic domain of kinase A affecting the ATP/ADP reactions and not the phosphotransferase functions of this domain. The inhibitory activity of KipI is counteracted by the product of another gene in the operon, KipA. This protein may bind to KipI, preventing its function as an inhibitor of kinase A. KipI may be the first representative of a new class of signal transduction inhibitors that function by direct interaction with the catalytic domain of histidine kinases to counteract signals influencing the "sensor" domain of such kinases. This inhibitor represents yet another way by which the phosphorelay signal transduction system is affected by negative regulators under the control of metabolic, environmental, or cell cycle influences antithetical to the initiation of developmental transcription. PMID- 9334323 TI - Taking a new TAK on tat transactivation. PMID- 9334322 TI - Topoisomerase IV, not gyrase, decatenates products of site-specific recombination in Escherichia coli. AB - DNA replication and recombination generate intertwined DNA intermediates that must be decatenated for chromosome segregation to occur. We showed recently that topoisomerase IV (topo IV) is the only important decatenase of DNA replication intermediates in bacteria. Earlier results, however, indicated that DNA gyrase has the primary role in unlinking the catenated products of site-specific recombination. To address this discordance, we constructed a set of isogenic strains that enabled us to inhibit selectively with the quinolone norfloxacin topo IV, gyrase, both enzymes, or neither enzyme in vivo. We obtained identical results for the decatenation of the products of two different site-specific recombination enzymes, phage lambda integrase and transposon Tn3 resolvase. Norfloxacin blocked decatenation in wild-type strains, but had no effect in strains with drug-resistance mutations in both gyrase and topo IV. When topo IV alone was inhibited, decatenation was almost completely blocked. If gyrase alone were inhibited, most of the catenanes were unlinked. We showed that topo IV is the primary decatenase in vivo and that this function is dependent on the level of DNA supercoiling. We conclude that the role of gyrase in decatenation is to introduce negative supercoils into DNA, which makes better substrates for topo IV. We also discovered that topo IV has an unexpectedly strong DNA relaxation activity that, together with gyrase and topo I, is able to set the supercoiling levels in Escherichia coli. PMID- 9334324 TI - Meiotic chromosomes: it takes two to tango. PMID- 9334325 TI - Transcription elongation factor P-TEFb is required for HIV-1 tat transactivation in vitro. AB - P-TEFb is a key regulator of the process controlling the processivity of RNA polymerase II and possesses a kinase activity that can phosphorylate the carboxy terminal domain of the largest subunit of RNA polymerase II. Here we report the cloning of the small subunit of Drosophila P-TEFb and the finding that it encodes a Cdc2-related protein kinase. Sequence comparison suggests that a protein with 72% identity, PITALRE, could be the human homolog of the Drosophila protein. Functional homology was suggested by transcriptional analysis of an RNA polymerase II promoter with HeLa nuclear extract depleted of PITALRE. Because the depleted extract lost the ability to produce long DRB-sensitive transcripts and this loss was reversed by the addition of purified Drosophila P-TEFb, we propose that PITALRE is a component of human P-TEFb. In addition, we found that PITALRE associated with the activation domain of HIV-1 Tat, indicating that P-TEFb is a Tat-associated kinase (TAK). An in vitro transcription assay demonstrates that the effect of Tat on transcription elongation requires P-TEFb and suggests that the enhancement of transcriptional processivity by Tat is attributable to enhanced function of P-TEFb on the HIV-1 LTR. PMID- 9334326 TI - P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro. AB - To identify novel inhibitors of transcriptional activation by the HIV Tat protein, we used a combination of in vitro and in vivo Tat-dependent transcription assays to screen >100,000 compounds. All compounds identified blocked Tat-dependent stimulation of transcriptional elongation. Analysis of a panel of structurally diverse inhibitors indicated that their target is the human homolog of Drosophila positive transcription elongation factor b (P-TEFb). Loss of Tat transactivation in extracts depleted of the kinase subunit of human P TEFb, PITALRE, was reversed by addition of partially purified human P-TEFb. Transfection experiments with wild-type or kinase knockout PITALRE demonstrated that P-TEFb is required for Tat function. Our results suggest that P-TEFb represents an attractive target for the development of novel HIV therapeutics. PMID- 9334329 TI - The Nun protein of bacteriophage HK022 inhibits translocation of Escherichia coli RNA polymerase without abolishing its catalytic activities. AB - Bacteriophage HK022 Nun protein blocks transcription elongation by Escherichia coli RNA polymerase in vitro without dissociating the transcription complex. Nun is active on complexes located at any template site tested. Ultimately, only the 3'-OH terminal nucleotide of the nascent transcript in an arrested complex can turn over; it is removed by pyrophosphate and restored with NTPs. This suggests that Nun inhibits the translocation of RNA polymerase without abolishing its catalytic activities. Unlike spontaneously arrested complexes, Nun-arrested complexes cannot be reactivated by transcription factor GreB. The various complexes show distinct patterns of nucleotide incorporation and pyrophosphorolysis before or after treatment with Nun, suggesting that the configuration of RNAP, transcript, and template DNA is different in each complex. PMID- 9334327 TI - The HIV transactivator TAT binds to the CDK-activating kinase and activates the phosphorylation of the carboxy-terminal domain of RNA polymerase II. AB - The human immunodeficiency virus encodes the transcriptional transactivator Tat, which binds to the transactivation response (TAR) RNA stem-loop in the viral long terminal repeat (LTR) and increases rates of elongation rather than initiation of transcription by RNA polymerase II (Pol II). In this study, we demonstrate that Tat binds directly to the cyclin-dependent kinase 7 (CDK7), which leads to productive interactions between Tat and the CDK-activating kinase (CAK) complex and between Tat and TFIIH. Tat activates the phosphorylation of the carboxy terminal domain (CTD) of Pol II by CAK in vitro. The ability of CAK to phosphorylate the CTD can be inhibited specifically by a CDK7 pseudosubstrate peptide that also inhibits transcriptional activation by Tat in vitro and in vivo. We conclude that the phosphorylation of the CTD by CAK is essential for Tat transactivation. Our data identify a cellular protein that interacts with the activation domain of Tat, demonstrate that this interaction is critical for the function of Tat, and provide a mechanism by which Tat increases the processivity of Pol II. PMID- 9334328 TI - TAFII250-dependent transcription of cyclin A is directed by ATF activator proteins. AB - A specific mutation in TAFII250, the largest subunit of the transcription factor TFIID, disrupts cell growth control in the temperature-sensitive mutant hamster cell line ts13. Transcription from the cyclin A and D1 but not the c-fos and myc promoters is also dramatically reduced in ts13 cells at the nonpermissive temperature. These findings provide an intriguing link between TAF-mediated transcriptional regulation and cell cycle progression. Here we report the mapping of an enhancer element in the cyclin A promoter (TSRE) that responds to mutations in TAFII250. An analysis of chimeric promoter constructs reveals that the cyclin A TSRE can confer TAFII250 dependence to the core promoter of c-fos. In addition, reciprocal hybrid promoter constructs suggest that TAFII250 also contributes to the transcriptional properties of the cyclin A core promoter. We have purified and identified cellular activators that specifically bind to the TSRE and mediate transcription in a TAFII250-dependent manner. By micropeptide sequencing, we determined that TSRE-binding proteins include members of the activating transcription factor (ATF) family. These results suggest that the ts13 mutation of TAFII250 has compromised the ability of TFIID to mediate activation of transcription by specific enhancer factors such as ATF, as well as to perform certain core promoter functions. These defects in TAFII250 apparently result in the down-regulation of key molecules, such as cyclin A, which may be responsible for the ts13 cell cycle arrest phenotype. PMID- 9334330 TI - The DAF-3 Smad protein antagonizes TGF-beta-related receptor signaling in the Caenorhabditis elegans dauer pathway. AB - Signals from TGF-beta superfamily receptors are transduced to the nucleus by Smad proteins, which transcriptionally activate target genes. In Caenorhabditis elegans, defects in a TGF-beta-related pathway cause a reversible developmental arrest and metabolic shift at the dauer larval stage. Null mutations in daf-3 suppress mutations in genes encoding this TGF-beta signal, its receptors, and associated Smad signal transduction proteins. daf-3 encodes a Smad protein that is most closely related to mammalian DPC4, and is expressed throughout development in many of the tissues that are remodeled during dauer development. DAF-4, the type II TGF-beta receptor in this pathway, is also expressed in remodeled tissues. These data suggest that the DAF-7 signal from sensory neurons acts as a neuroendocrine signal throughout the body to directly regulate developmental and metabolic shifts in tissues that are remodeled during dauer formation. A full-length functional DAF-3/GFP fusion protein is predominantly cytoplasmic, and this localization is independent of activity of the upstream TGF beta-related pathway. However, this fusion protein is associated with chromosomes in mitotic cells, suggesting that DAF-3 binds DNA directly or indirectly. DAF-3 transgenes also interfere with dauer formation, perhaps attributable to a dosage effect. A truncated DAF-3/GFP fusion protein that is predominantly nuclear interferes with dauer formation, implying a role for DAF-3 in the nucleus. These data suggest that DAF-7 signal transduction antagonizes or modifies DAF-3 Smad activity in the nucleus to induce reproductive development; when DAF-7 signals are disabled, unmodified DAF-3 Smad activity mediates dauer arrest and its associated metabolic shift. Therefore, daf-3 is unique in that it is antagonized, rather than activated, by a TGF-beta pathway. PMID- 9334331 TI - The Drosophila neuregulin homolog Vein mediates inductive interactions between myotubes and their epidermal attachment cells. AB - Inductive interactions between cells of distinct fates underlie the basis for morphogenesis and organogenesis across species. In the Drosophila embryo, somatic myotubes form specific interactions with their epidermal muscle attachment (EMA) cells. The establishment of these interactions is a first step toward further differentiation of the EMA cells into elongated tendon cells containing an organized array of microtubules and microfilaments. Here we show that the molecular signal for terminal differentiation of tendon cells is the secreted Drosophila neuregulin-like growth factor Vein produced by the myotubes. Although vein mRNA is produced by all of the myotubes, Vein protein is secreted and accumulates specifically at the muscle-tendon cell junctional site. In loss-of function vein mutant embryos, muscle-dependent differentiation of tendon cells, measured by the level of expression of specific markers (Delilah and beta1 tubulin) is blocked. When Vein is expressed in ectopic ectodermal cells, it induces the ectopic expression of these genes. Our results favor the possibility that the Drosophila EGF receptor DER/Egfr expressed by the EMA cells functions as a receptor for Vein. We show that Vein/Egfr binding activates the Ras pathway in the EMA cells leading to the transcription of the tendon-specific genes, stripe, delilah, and beta1 tubulin. In Egfr1F26 mutant embryos that lack functional Egfr expression, the levels of Delilah and beta1 tubulin are very low. In addition, the ability of ectopic Vein to induce the expression of Delilah and beta1 tubulin depends on the presence of functional Egfrs. Finally, activation of the Egfr signaling pathway by either ectopically secreted Spitz, or activated Ras, leads to the ectopic expression of Delilah. These results suggest that inductive interactions between myotubes and their epidermal muscle attachment cells are initiated by the binding of Vein, to the Egfr on the surface of EMA cells. PMID- 9334332 TI - Mammalian homologs of seven in absentia regulate DCC via the ubiquitin-proteasome pathway. AB - DCC (deleted in colorectal cancer) is postulated to function as transmembrane receptor for the axon and cell guidance factor netrin-1. We report here that the DCC cytoplasmic domain binds to proteins encoded by mammalian homologs of the Drosophila seven in absentia (sina) gene, as well as Drosophila Sina. Sina has a critical role in R7 photoreceptor development and shows upward of 85% amino acid identity with its mammalian homologs (termed Siahs), but the function of the Sina/Siah proteins has not been defined. We sought, therefore, to characterize further their interaction with DCC. Immunofluorescence studies suggested the Sina/Siah proteins localized predominantly in the cytoplasm and in association with DCC. DCC was found to be ubiquitinated and the Sina/Siah proteins regulated its expression. Proteasome inhibitors blocked the effects of Sina/Siah on DCC, and the Sina/Siah proteins interacted with ubiquitin-conjugating enzymes (Ubcs). A mutant Siah protein lacking the amino-terminal Ubc-binding sequences complexed with DCC, but did not degrade it. The in vivo interaction between Sina/Siah and DCC was confirmed through studies of transgenic Drosophila lines in which DCC and Sina were ectopically expressed in the eye. Taken together, the data imply that the Sina/Siah proteins regulate DCC and perhaps other proteins via the ubiquitin proteasome pathway. PMID- 9334333 TI - Inhibition of Caenorhabditis elegans vulval induction by gap-1 and by let-23 receptor tyrosine kinase. AB - During induction of the Caenorhabditis elegans hermaphrodite vulva, a signal from the anchor cell activates the LET-23 epidermal growth factor receptor (EGFR)/LET 60 Ras/MPK-1 MAP kinase signaling pathway in the vulval precursor cells. We have characterized two mechanisms that limit the extent of vulval induction. First, we found that gap-1 may directly inhibit the LET-60 Ras signaling pathway. We identified the gap-1 gene in a genetic screen for inhibitors of vulval induction. gap-1 is predicted to encode a protein similar to GTPase-activating proteins that likely functions to inhibit the signaling activity of LET-60 Ras. A loss-of function mutation in gap-1 suppresses the vulvaless phenotype of mutations in the let-60 ras signaling pathway, but a gap-1 single mutant does not exhibit excess vulval induction. Second, we found that let-23 EGFR prevents vulval induction in a cell-nonautonomous manner, in addition to its cell-autonomous role in activating the let-60 ras/mpk-1 signaling pathway. Using genetic mosaic analysis, we show that let-23 activity in the vulval precursor cell closest to the anchor cell (P6.p) prevents induction of vulval precursor cells further away from the anchor cell (P3.p, P4.p, and P8.p). This result suggests that LET-23 in proximal vulval precursor cells might bind and sequester the inductive signal LIN-3 EGF, thereby preventing diffusion of the inductive signal to distal vulval precursor cells. PMID- 9334335 TI - Alternative 3'-end processing of U5 snRNA by RNase III. AB - The cellular components required to form the 3' ends of small nuclear RNAs are unknown. U5 snRNA from Saccharomyces cerevisiae is found in two forms that differ in length at their 3' ends (U5L and U5S). When added to a yeast cell free extract, synthetic pre-U5 RNA bearing downstream genomic sequences is processed efficiently and accurately to generate both mature forms of U5. The two forms of U5 are produced in vitro by alternative 3'-end processing. A temperature sensitive mutation in the RNT1 gene encoding RNase III blocks accumulation of U5L in vivo. In vitro, alternative cleavage of the U5 precursor by RNase III determines the choice between the two multistep pathways that lead to U5L and U5S, one of which (U5L) is strictly dependent on RNase III. These results identify RNase III as a trans-acting factor involved in 3'-end formation of snRNA and show how RNase III might regulate alternative RNA processing pathways. PMID- 9334334 TI - Chip, a widely expressed chromosomal protein required for segmentation and activity of a remote wing margin enhancer in Drosophila. AB - The mechanisms allowing remote enhancers to regulate promoters several kilobase pairs away are unknown but are blocked by the Drosophila suppressor of Hairy-wing protein (Suhw) that binds to gypsy retrovirus insertions between enhancers and promoters. Suhw bound to a gypsy insertion in the cut gene also appears to act interchromosomally to antagonize enhancer-promoter interactions on the homologous chromosome when activity of the Chip gene is reduced. This implicates Chip in enhancer-promoter communication. We cloned Chip and find that it encodes a homolog of the recently discovered mouse Nli/Ldb1/Clim-2 and Xenopus Xldb1 proteins that bind nuclear LIM domain proteins. Chip protein interacts with the LIM domains in the Apterous homeodomain protein, and Chip interacts genetically with apterous, showing that these interactions are important for Apterous function in vivo. Importantly, Chip also appears to have broad functions beyond interactions with LIM domain proteins. Chip is present in all nuclei examined and at numerous sites along the salivary gland polytene chromosomes. Embryos without Chip activity lack segments and show abnormal gap and pair-rule gene expression, although no LIM domain proteins are known to regulate segmentation. We conclude that Chip is a ubiquitous chromosomal factor required for normal expression of diverse genes at many stages of development. We suggest that Chip cooperates with different LIM domain proteins and other factors to structurally support remote enhancer-promoter interactions. PMID- 9334336 TI - Lessons from mice without telomerase. PMID- 9334337 TI - Nucleocytoplasmic recycling of the nuclear localization signal receptor alpha subunit in vivo is dependent on a nuclear export signal, energy, and RCC1. AB - Nuclear protein import requires a nuclear localization signal (NLS) receptor and at least three other cytoplasmic factors. The alpha subunit of the NLS receptor, Rag cohort 1 (Rch1), enters the nucleus, probably in a complex with the beta subunit of the receptor, as well as other import factors and the import substrate. To learn more about which factors and/or events end the import reaction and how the import factors return to the cytoplasm, we have studied nucleocytoplasmic shuttling of Rch1 in vivo. Recombinant Rch1 microinjected into Vero or tsBN2 cells was found primarily in the cytoplasm. Rch1 injected into the nucleus was rapidly exported in a temperature-dependent manner. In contrast, a mutant of Rch1 lacking the first 243 residues accumulated in the nuclei of Vero cells after cytoplasmic injection. After nuclear injection, the truncated Rch1 was retained in the nucleus, but either Rch1 residues 207-217 or a heterologous nuclear export signal, but not a mutant form of residues 207-217, restored nuclear export. Loss of the nuclear transport factor RCC1 (regulator of chromosome condensation) at the nonpermissive temperature in the thermosensitive mutant cell line tsBN2 caused nuclear accumulation of wild-type Rch1 injected into the cytoplasm. However, free Rch1 injected into nuclei of tsBN2 cells at the nonpermissive temperature was exported. These results suggested that RCC1 acts at an earlier step in Rch1 recycling, possibly the disassembly of an import complex that contains Rch1 and the import substrate. Consistent with this possibility, incubation of purified RanGTP and RCC1 with NLS receptor and import substrate prevented assembly of receptor/substrate complexes or stimulated their disassembly. PMID- 9334338 TI - p28 Bap31, a Bcl-2/Bcl-XL- and procaspase-8-associated protein in the endoplasmic reticulum. AB - We have identified a human Bcl-2-interacting protein, p28 Bap31. It is a 28-kD (p28) polytopic integral protein of the endoplasmic reticulum whose COOH-terminal cytosolic region contains overlapping predicted leucine zipper and weak death effector homology domains, flanked on either side by identical caspase recognition sites. In cotransfected 293T cells, p28 is part of a complex that includes Bcl-2/Bcl-XL and procaspase-8 (pro-FLICE). Bax, a pro-apoptotic member of the Bcl-2 family, does not associate with the complex; however, it prevents Bcl-2 from doing so. In the absence (but not presence) of elevated Bcl-2 levels, apoptotic signaling by adenovirus E1A oncoproteins promote cleavage of p28 at the two caspase recognition sites. Purified caspase-8 (FLICE/MACH/Mch5) and caspase 1(ICE), but not caspase-3 (CPP32/apopain/ Yama), efficiently catalyze this reaction in vitro. The resulting NH2-terminal p20 fragment induces apoptosis when expressed ectopically in otherwise normal cells. Taken together, the results suggest that p28 Bap31 is part of a complex in the endoplasmic reticulum that mechanically bridges an apoptosis-initiating caspase, like procaspase-8, with the anti-apoptotic regulator Bcl-2 or Bcl-XL. This raises the possibility that the p28 complex contributes to the regulation of procaspase-8 or a related caspase in response to E1A, dependent on the status of the Bcl-2 setpoint within the complex. PMID- 9334339 TI - Phosphatidylinositol 3-kinase is required for the formation of constitutive transport vesicles from the TGN. AB - An 85-kD cytosolic complex (p62(cplx)), consisting of a 62-kD phosphoprotein (p62) and a 25-kD GTPase, has been shown to be essential for the cell-free reconstitution of polymeric IgA receptor (pIgA-R)-containing exocytic transport vesicle formation from the TGN (Jones, S.M., J.R. Crosby, J. Salamero, and K.E. Howell. 1993. J. Cell Biol. 122:775-788). Here the p62(cplx) is identified as a regulatory subunit of a novel phosphatidylinositol 3-kinase (PI3-kinase). This p62(cplx)-associated PI3-kinase activity is stimulated by activation of the p62(cplx)-associated GTPase, and is specific for phosphatidylinositol (PI) as substrate, and is sensitive to wortmannin at micromolar concentrations. The direct role of this p62(cplx)-associated PI3-kinase activity in TGN-derived vesicle formation is indicated by the finding that both lipid kinase activity and the formation of pIgA-R-containing exocytic vesicles from the TGN are inhibited by wortmannin with similar dose-response curves and 50% inhibitory concentrations (3.5 microM). These findings indicate that phosphatidylinositol-3-phosphate (PI[3]P) is required for the formation of TGN-derived exocytic transport vesicles, and that the p62(cplx)-associated PI3-kinase and an activated GTPase are the essential molecules that drive production of this PI(3)P. PMID- 9334340 TI - The phosphatidylinositol transfer protein domain of Drosophila retinal degeneration B protein is essential for photoreceptor cell survival and recovery from light stimulation. AB - The Drosophila retinal degeneration B (rdgB) gene encodes an integral membrane protein involved in phototransduction and prevention of retinal degeneration. RdgB represents a nonclassical phosphatidylinositol transfer protein (PITP) as all other known PITPs are soluble polypeptides. Our data demonstrate roles for RdgB in proper termination of the phototransduction light response and dark recovery of the photoreceptor cells. Expression of RdgB's PITP domain as a soluble protein (RdgB-PITP) in rdgB2 mutant flies is sufficient to completely restore the wild-type electrophysiological light response and prevent the degeneration. However, introduction of the T59E mutation, which does not affect RdgB-PITP's phosphatidylinositol (PI) and phosphatidycholine (PC) transfer in vitro, into the soluble (RdgB-PITP-T59E) or full-length (RdgB-T59E) proteins eliminated rescue of retinal degeneration in rdgB2 flies, while the light response was partially maintained. Substitution of the rat brain PITPalpha, a classical PI transfer protein, for RdgB's PITP domain (PITPalpha or PITPalpha RdgB chimeric protein) neither restored the light response nor maintained retinal integrity when expressed in rdgB2 flies. Therefore, the complete repertoire of essential RdgB functions resides in RdgB's PITP domain, but other PITPs possessing PI and/or PC transfer activity in vitro cannot supplant RdgB function in vivo. Expression of either RdgB-T59E or PITPalpha-RdgB in rdgB+ flies produced a dominant retinal degeneration phenotype. Whereas RdgB-T59E functioned in a dominant manner to significantly reduce steady-state levels of rhodopsin, PITPalpha-RdgB was defective in the ability to recover from prolonged light stimulation and caused photoreceptor degeneration through an unknown mechanism. This in vivo analysis of PITP function in a metazoan system provides further insights into the links between PITP dysfunction and an inherited disease in a higher eukaryote. PMID- 9334341 TI - Dynamic distribution of chemoattractant receptors in living cells during chemotaxis and persistent stimulation. AB - While the localization of chemoattractant receptors on randomly oriented cells has been previously studied by immunohistochemistry, the instantaneous distribution of receptors on living cells undergoing directed migration has not been determined. To do this, we replaced cAR1, the primary cAMP receptor of Dictyostelium, with a cAR1-green fluorescence protein fusion construct. We found that this chimeric protein is functionally indistinguishable from wild-type cAR1. By time-lapse imaging of single cells, we observed that the receptors remained evenly distributed on the cell surface and all of its projections during chemotaxis involving turns and reversals of polarity directed by repositioning of a chemoattractant-filled micropipet. Thus, cell polarization cannot result from a gradient-induced asymmetric distribution of chemoattractant receptors. Some newly extended pseudopods at migration fronts showed a transient drop in fluorescence signals, suggesting that the flow of receptors into these zones may slightly lag behind the protrusion process. Challenge with a uniform increase in chemoattractant, sufficient to cause a dramatic decrease in the affinity of surface binding sites and cell desensitization, also did not significantly alter the distribution profile. Hence, the induced reduction in binding activity and cellular sensitivity cannot be due to receptor relocalization. The chimeric receptors were able to "cap" rapidly during treatment with Con A, suggesting that they are mobile in the plane of the cell membrane. This capping was not influenced by pretreatment with chemoattractant. PMID- 9334342 TI - Animal models for muscular dystrophy show different patterns of sarcolemmal disruption. AB - Genetic defects in a number of components of the dystrophin-glycoprotein complex (DGC) lead to distinct forms of muscular dystrophy. However, little is known about how alterations in the DGC are manifested in the pathophysiology present in dystrophic muscle tissue. One hypothesis is that the DGC protects the sarcolemma from contraction-induced damage. Using tracer molecules, we compared sarcolemmal integrity in animal models for muscular dystrophy and in muscular dystrophy patient samples. Evans blue, a low molecular weight diazo dye, does not cross into skeletal muscle fibers in normal mice. In contrast, mdx mice, a dystrophin deficient animal model for Duchenne muscular dystrophy, showed significant Evans blue accumulation in skeletal muscle fibers. We also studied Evans blue dispersion in transgenic mice bearing different dystrophin mutations, and we demonstrated that cytoskeletal and sarcolemmal attachment of dystrophin might be a necessary requirement to prevent serious fiber damage. The extent of dye incorporation in transgenic mice correlated with the phenotypic severity of similar dystrophin mutations in humans. We furthermore assessed Evans blue incorporation in skeletal muscle of the dystrophia muscularis (dy/dy) mouse and its milder allelic variant, the dy2J/dy2J mouse, animal models for congenital muscular dystrophy. Surprisingly, these mice, which have defects in the laminin alpha2-chain, an extracellular ligand of the DGC, showed little Evans blue accumulation in their skeletal muscles. Taken together, these results suggest that the pathogenic mechanisms in congenital muscular dystrophy are different from those in Duchenne muscular dystrophy, although the primary defects originate in two components associated with the same protein complex. PMID- 9334344 TI - Analysis of the actin-myosin II system in fish epidermal keratocytes: mechanism of cell body translocation. AB - While the protrusive event of cell locomotion is thought to be driven by actin polymerization, the mechanism of forward translocation of the cell body is unclear. To elucidate the mechanism of cell body translocation, we analyzed the supramolecular organization of the actin-myosin II system and the dynamics of myosin II in fish epidermal keratocytes. In lamellipodia, long actin filaments formed dense networks with numerous free ends in a brushlike manner near the leading edge. Shorter actin filaments often formed T junctions with longer filaments in the brushlike area, suggesting that new filaments could be nucleated at sides of preexisting filaments or linked to them immediately after nucleation. The polarity of actin filaments was almost uniform, with barbed ends forward throughout most of the lamellipodia but mixed in arc-shaped filament bundles at the lamellipodial/cell body boundary. Myosin II formed discrete clusters of bipolar minifilaments in lamellipodia that increased in size and density towards the cell body boundary and colocalized with actin in boundary bundles. Time-lapse observation demonstrated that myosin clusters appeared in the lamellipodia and remained stationary with respect to the substratum in locomoting cells, but they exhibited retrograde flow in cells tethered in epithelioid colonies. Consequently, both in locomoting and stationary cells, myosin clusters approached the cell body boundary, where they became compressed and aligned, resulting in the formation of boundary bundles. In locomoting cells, the compression was associated with forward displacement of myosin features. These data are not consistent with either sarcomeric or polarized transport mechanisms of cell body translocation. We propose that the forward translocation of the cell body and retrograde flow in the lamellipodia are both driven by contraction of an actin myosin network in the lamellipodial/cell body transition zone. PMID- 9334343 TI - Evidence for a conformational change in actin induced by fimbrin (N375) binding. AB - Fimbrin belongs to a superfamily of actin cross-linking proteins that share a conserved 27-kD actin-binding domain. This domain contains a tandem duplication of a sequence that is homologous to calponin. Calponin homology (CH) domains not only cross-link actin filaments into bundles and networks, but they also bind intermediate filaments and some signal transduction proteins to the actin cytoskeleton. This fundamental role of CH domains as a widely used actin-binding domain underlines the necessity to understand their structural interaction with actin. Using electron cryomicroscopy, we have determined the three-dimensional structure of F-actin and F-actin decorated with the NH2-terminal CH domains of fimbrin (N375). In a difference map between actin filaments and N375-decorated actin, one end of N375 is bound to a concave surface formed between actin subdomains 1 and 2 on two neighboring actin monomers. In addition, a fit of the atomic model for the actin filament to the maps reveals the actin residues that line, the binding surface. The binding of N375 changes actin, which we interpret as a movement of subdomain 1 away from the bound N375. This change in actin structure may affect its affinity for other actin-binding proteins and may be part of the regulation of the cytoskeleton itself. Difference maps between actin and actin decorated with other proteins provides a way to look for novel structural changes in actin. PMID- 9334345 TI - Actomyosin-based retrograde flow of microtubules in the lamella of migrating epithelial cells influences microtubule dynamic instability and turnover and is associated with microtubule breakage and treadmilling. AB - We have discovered several novel features exhibited by microtubules (MTs) in migrating newt lung epithelial cells by time-lapse imaging of fluorescently labeled, microinjected tubulin. These cells exhibit leading edge ruffling and retrograde flow in the lamella and lamellipodia. The plus ends of lamella MTs persist in growth perpendicular to the leading edge until they reach the base of the lamellipodium, where they oscillate between short phases of growth and shortening. Occasionally "pioneering" MTs grow into the lamellipodium, where microtubule bending and reorientation parallel to the leading edge is associated with retrograde flow. MTs parallel to the leading edge exhibit significantly different dynamics from MTs perpendicular to the cell edge. Both parallel MTs and photoactivated fluorescent marks on perpendicular MTs move rearward at the 0.4 mircon/min rate of retrograde flow in the lamella. MT rearward transport persists when MT dynamic instability is inhibited by 100-nM nocodazole but is blocked by inhibition of actomyosin by cytochalasin D or 2,3-butanedione-2-monoxime. Rearward flow appears to cause MT buckling and breaking in the lamella. 80% of free minus ends produced by breakage are stable; the others shorten and pause, leading to MT treadmilling. Free minus ends of unknown origin also depolymerize into the field of view at the lamella. Analysis of MT dynamics at the centrosome shows that these minus ends do not arise by centrosomal ejection and that approximately 80% of the MTs in the lamella are not centrosome bound. We propose that actomyosin-based retrograde flow of MTs causes MT breakage, forming quasi stable noncentrosomal MTs whose turnover is regulated primarily at their minus ends. PMID- 9334346 TI - The microtubule-dependent motor centromere-associated protein E (CENP-E) is an integral component of kinetochore corona fibers that link centromeres to spindle microtubules. AB - Centromere-associated protein E (CENP-E) is a kinesin-related microtubule motor protein that is essential for chromosome congression during mitosis. Using immunoelectron microscopy, CENP-E is shown to be an integral component of the kinetochore corona fibers that tether centromeres to the spindle. Immediately upon nuclear envelope fragmentation, an associated plus end motor trafficks cytoplasmic CENP-E toward chromosomes along astral microtubules that enter the nuclear volume. Before or concurrently with initial lateral attachment of spindle microtubules, CENP-E targets to the outermost region of the developing kinetochores. After stable attachment, throughout chromosome congression, at metaphase, and throughout anaphase A, CENP-E is a constituent of the corona fibers, extending at least 50 nm away from the kinetochore outer plate and intertwining with spindle microtubules. In congressing chromosomes, CENP-E is preferentially associated with (or accessible at) the stretched, leading kinetochore known to provide the primary power for chromosome movement. Taken together, this evidence strongly supports a model in which CENP-E functions in congression to tether kinetochores to the disassembling microtubule plus ends. PMID- 9334347 TI - Thrombopoietin-induced polyploidization of bone marrow megakaryocytes is due to a unique regulatory mechanism in late mitosis. AB - Megakaryocytes undergo a unique differentiation program, becoming polyploid through repeated cycles of DNA synthesis without concomitant cell division. However, the mechanism underlying this polyploidization remains totally unknown. It has been postulated that polyploidization is due to a skipping of mitosis after each round of DNA replication. We carried out immunohistochemical studies on mouse bone marrow megakaryocytes during thrombopoietin- induced polyploidization and found that during this process megakaryocytes indeed enter mitosis and progress through normal prophase, prometaphase, metaphase, and up to anaphase A, but not to anaphase B, telophase, or cytokinesis. It was clearly observed that multiple spindle poles were formed as the polyploid megakaryocytes entered mitosis; the nuclear membrane broke down during prophase; the sister chromatids were aligned on a multifaced plate, and the centrosomes were symmetrically located on either side of each face of the plate at metaphase; and a set of sister chromatids moved into the multiple centrosomes during anaphase A. We further noted that the pair of spindle poles in anaphase were located in close proximity to each other, probably because of the lack of outward movement of spindle poles during anaphase B. Thus, the reassembling nuclear envelope may enclose all the sister chromatids in a single nucleus at anaphase and then skip telophase and cytokinesis. These observations clearly indicate that polyploidization of megakaryocytes is not simply due to a skipping of mitosis, and that the megakaryocytes must have a unique regulatory mechanism in anaphase, e.g., factors regulating anaphase such as microtubule motor proteins might be involved in this polyploidization process. PMID- 9334348 TI - The yeast motor protein, Kar3p, is essential for meiosis I. AB - The recognition and alignment of homologous chromosomes early in meiosis is essential for their subsequent segregation at anaphase I; however, the mechanism by which this occurs is unknown. We demonstrate here that, in the absence of the molecular motor, Kar3p, meiotic cells are blocked with prophase monopolar microtubule arrays and incomplete synaptonemal complex (SC) formation. kar3 mutants exhibit very low levels of heteroallelic recombination. kar3 mutants do produce double-strand breaks that act as initiation sites for meiotic recombination in yeast, but at levels severalfold reduced from wild-type. These data are consistent with a meiotic role for Kar3p in the events that culminate in synapsis of homologues. PMID- 9334350 TI - Increased apoptosis arising from increased expression of the Alzheimer's disease associated presenilin-2 mutation (N141I). AB - Mutations in the genes for presenilin 1 and 2 (PS-1 and PS-2) have been linked to development of early-onset Alzheimer's disease (AD). As neither the normal function of either presenilin is known nor why mutations cause disease, we examined the properties of wild-type, truncated, and mutant PS-2 upon expression in HeLa cells. Although HeLa cells are strongly predisposed to continued mitosis, expression of PS-2 induced programmed cell death (apoptosis). Direct evidence for apoptosis was obtained by double staining for terminal deoxynucleotide transferase nick end labeling (TUNEL) and PS-2 expression and by following green fluorescent protein-tagged PS-2 over time. Deletion analysis indicates that as little as 166 NH2-terminal residues of PS-2 are sufficient for endoplasmic reticulum (ER) localization and apoptosis. Moreover, the AD- associated PS-2 missense mutation (N141I) more efficiently induced cell death compared to wild type PS-2 despite lower mutant protein accumulation. Expression of the presenilins in several other cell lines and transgenic mice has been accompanied by rapid protein cleavage without the induction of cell death. In contrast, PS-2 expressed in HeLa cells was not cleaved, and cell death occurred. We hypothesize that full-length but not cleaved PS-2 may be important in the regulation or induction of apoptosis. PMID- 9334349 TI - Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution. AB - Dynactin is a multisubunit complex that plays an accessory role in cytoplasmic dynein function. Overexpression in mammalian cells of one dynactin subunit, dynamitin, disrupts the complex, resulting in dissociation of cytoplasmic dynein from prometaphase kinetochores, with consequent perturbation of mitosis (Echeverri, C.J., B.M. Paschal, K.T. Vaughan, and R.B. Vallee. 1996. J. Cell Biol. 132:617-634). Based on these results, dynactin was proposed to play a role in linking cytoplasmic dynein to kinetochores and, potentially, to membrane organelles. The current study reports on the dynamitin interphase phenotype. In dynamitin-overexpressing cells, early endosomes (labeled with antitransferrin receptor), as well as late endosomes and lysosomes (labeled with anti-lysosome associated membrane protein-1 [LAMP-1]), were redistributed to the cell periphery. This redistribution was disrupted by nocodazole, implicating an underlying plus end-directed microtubule motor activity. The Golgi stack, monitored using sialyltransferase, galactosyltransferase, and N acetylglucosaminyltransferase I, was dramatically disrupted into scattered structures that colocalized with components of the intermediate compartment (ERGIC-53 and ERD-2). The disrupted Golgi elements were revealed by EM to represent short stacks similar to those formed by microtubule-depolymerizing agents. Golgi-to-ER traffic of stack markers induced by brefeldin A was not inhibited by dynamitin overexpression. Time-lapse observations of dynamitin overexpressing cells recovering from brefeldin A treatment revealed that the scattered Golgi elements do not undergo microtubule-based transport as seen in control cells, but rather, remain stationary at or near their ER exit sites. These results indicate that dynactin is specifically required for ongoing centripetal movement of endocytic organelles and components of the intermediate compartment. Results similar to those of dynamitin overexpression were obtained by microinjection with antidynein intermediate chain antibody, consistent with a role for dynactin in mediating interactions of cytoplasmic dynein with specific membrane organelles. These results suggest that dynamitin plays a pivotal role in regulating organelle movement at the level of motor-cargo binding. PMID- 9334351 TI - ATP- and gap junction-dependent intercellular calcium signaling in osteoblastic cells. AB - Many cells coordinate their activities by transmitting rises in intracellular calcium from cell to cell. In nonexcitable cells, there are currently two models for intercellular calcium wave propagation, both of which involve release of inositol trisphosphate (IP3)- sensitive intracellular calcium stores. In one model, IP3 traverses gap junctions and initiates the release of intracellular calcium stores in neighboring cells. Alternatively, calcium waves may be mediated not by gap junctional communication, but rather by autocrine activity of secreted ATP on P2 purinergic receptors. We studied mechanically induced calcium waves in two rat osteosarcoma cell lines that differ in the gap junction proteins they express, in their ability to pass microinjected dye from cell to cell, and in their expression of P2Y2 (P2U) purinergic receptors. ROS 17/2.8 cells, which express the gap junction protein connexin43 (Cx43), are well dye coupled, and lack P2U receptors, transmitted slow gap junction-dependent calcium waves that did not require release of intracellular calcium stores. UMR 106-01 cells predominantly express the gap junction protein connexin 45 (Cx45), are poorly dye coupled, and express P2U receptors; they propagated fast calcium waves that required release of intracellular calcium stores and activation of P2U purinergic receptors, but not gap junctional communication. ROS/P2U transfectants and UMR/Cx43 transfectants expressed both types of calcium waves. Gap junction independent, ATP-dependent intercellular calcium waves were also seen in hamster tracheal epithelia cells. These studies demonstrate that activation of P2U purinergic receptors can propagate intercellular calcium, and describe a novel Cx43-dependent mechanism for calcium wave propagation that does not require release of intracellular calcium stores by IP3. These studies suggest that gap junction communication mediated by either Cx43 or Cx45 does not allow passage of IP3 well enough to elicit release of intracellular calcium stores in neighboring cells. PMID- 9334352 TI - Actinin-associated LIM protein: identification of a domain interaction between PDZ and spectrin-like repeat motifs. AB - PDZ motifs are protein-protein interaction domains that often bind to COOH terminal peptide sequences. The two PDZ proteins characterized in skeletal muscle, syntrophin and neuronal nitric oxide synthase, occur in the dystrophin complex, suggesting a role for PDZ proteins in muscular dystrophy. Here, we identify actinin-associated LIM protein (ALP), a novel protein in skeletal muscle that contains an NH2-terminal PDZ domain and a COOH-terminal LIM motif. ALP is expressed at high levels only in differentiated skeletal muscle, while an alternatively spliced form occurs at low levels in the heart. ALP is not a component of the dystrophin complex, but occurs in association with alpha-actinin 2 at the Z lines of myofibers. Biochemical and yeast two-hybrid analyses demonstrate that the PDZ domain of ALP binds to the spectrin-like motifs of alpha actinin-2, defining a new mode for PDZ domain interactions. Fine genetic mapping studies demonstrate that ALP occurs on chromosome 4q35, near the heterochromatic locus that is mutated in fascioscapulohumeral muscular dystrophy. PMID- 9334354 TI - Induction of apoptosis after expression of PYK2, a tyrosine kinase structurally related to focal adhesion kinase. AB - Many cells (e.g., epithelial cells) require attachment to the extracellular matrix (ECM) to survive, a phenomenon known as anchorage-dependent cell survival. Disruption of the cell-ECM interactions mediated by the integrin receptors results in apoptosis. Focal adhesion kinase (FAK), a 125-kD protein tyrosine kinase activated by integrin engagement, appears to be involved in mediating cell attachment and survival. Proline-rich tyrosine kinase 2 (PYK2), also known as cellular adhesion kinase beta (CAKbeta) and related adhesion focal tyrosine kinase, is a second member of the FAK subfamily and is activated by an increase in intracellular calcium levels, or treatment with TNFalpha and UV light. However, the function of PYK2 remains largely unknown. In this study, we show that over-expression of PYK2, but not FAK, in rat and mouse fibroblasts leads to apoptotic cell death. Using a series of deletion mutants and chimeric fusion proteins of PYK2/FAK, we determined that the NH2-terminal domain and tyrosine kinase activity of PYK2 were required for the efficient induction of apoptosis. Furthermore, the apoptosis mediated by PYK2 could be suppressed by over expressing catalytically active v-Src, c-Src, phosphatidylinositol-3-kinase, or Akt/protein kinase B. In addition, it could also be suppressed by overexpressing an ICE or ICE-like proteinase inhibitor, crmA, but not Bcl2. Collectively, our results suggest that PYK2 and FAK, albeit highly homologous in primary structure, appear to have different functions; FAK is required for cell survival, whereas PYK2 induces apoptosis in fibroblasts. PMID- 9334353 TI - Afadin: A novel actin filament-binding protein with one PDZ domain localized at cadherin-based cell-to-cell adherens junction. AB - A novel actin filament (F-actin)-binding protein with a molecular mass of approximately 205 kD (p205), which was concentrated at cadherin-based cell-to cell adherens junction (AJ), was isolated and characterized. p205 was purified from rat brain and its cDNA was cloned from a rat brain cDNA library. p205 was a protein of 1,829 amino acids (aa) with a calculated molecular mass of 207,667 kD. p205 had one F-actin-binding domain at 1,631-1,829 aa residues and one PDZ domain at 1,016- 1,100 aa residues, a domain known to interact with transmembrane proteins. p205 was copurified from rat brain with another protein with a molecular mass of 190 kD (p190). p190 was a protein of 1,663 aa with a calculated molecular mass of 188,971 kD. p190 was a splicing variant of p205 having one PDZ domain at 1,009-1,093 aa residues but lacking the F-actin-binding domain. Homology search analysis revealed that the aa sequence of p190 showed 90% identity over the entire sequence with the product of the AF-6 gene, which was found to be fused to the ALL-1 gene, known to be involved in acute leukemia. p190 is likely to be a rat counterpart of human AF-6 protein. p205 bound along the sides of F-actin but hardly showed the F-actin-cross-linking activity. Northern and Western blot analyses showed that p205 was ubiquitously expressed in all the rat tissues examined, whereas p190 was specifically expressed in brain. Immunofluorescence and immunoelectron microscopic studies revealed that p205 was concentrated at cadherin-based cell-to-cell AJ of various tissues. We named p205 l-afadin (a large splicing variant of AF-6 protein localized at adherens junction) and p190 s-afadin (a small splicing variant of l-afadin). These results suggest that l-afadin serves as a linker of the actin cytoskeleton to the plasma membrane at cell-to-cell AJ. PMID- 9334355 TI - Expression of a truncated, kinase-defective TGF-beta type II receptor in mouse skeletal tissue promotes terminal chondrocyte differentiation and osteoarthritis. AB - Members of the TGF-beta superfamily are important regulators of skeletal development. TGF-betas signal through heteromeric type I and type II receptor serine/threonine kinases. When over-expressed, a cytoplasmically truncated type II receptor can compete with the endogenous receptors for complex formation, thereby acting as a dominant-negative mutant (DNIIR). To determine the role of TGF-betas in the development and maintenance of the skeleton, we have generated transgenic mice (MT-DNIIR-4 and -27) that express the DNIIR in skeletal tissue. DNIIR mRNA expression was localized to the periosteum/perichondrium, syno-vium, and articular cartilage. Lower levels of DNIIR mRNA were detected in growth plate cartilage. Transgenic mice frequently showed bifurcation of the xiphoid process and sternum. They also developed progressive skeletal degeneration, resulting by 4 to 8 mo of age in kyphoscoliosis and stiff and torqued joints. The histology of affected joints strongly resembled human osteo-arthritis. The articular surface was replaced by bone or hypertrophic cartilage as judged by the expression of type X collagen, a marker of hypertrophic cartilage normally absent from articular cartilage. The synovium was hyperplastic, and cartilaginous metaplasia was observed in the joint space. We then tested the hypothesis that TGF-beta is required for normal differentiation of cartilage in vivo. By 4 and 8 wk of age, the level of type X collagen was increased in growth plate cartilage of transgenic mice relative to wild-type controls. Less proteoglycan staining was detected in the growth plate and articular cartilage matrix of transgenic mice. Mice that express DNIIR in skeletal tissue also demonstrated increased Indian hedgehog (IHH) expression. IHH is a secreted protein that is expressed in chondrocytes that are committed to becoming hypertrophic. It is thought to be involved in a feedback loop that signals through the periosteum/ perichondrium to inhibit cartilage differentiation. The data suggest that TGF-beta may be critical for multifaceted maintenance of synovial joints. Loss of responsiveness to TGF beta promotes chondrocyte terminal differentiation and results in development of degenerative joint disease resembling osteoarthritis in humans. PMID- 9334356 TI - Laminin alpha1 chain synthesis in the mouse developing lung: requirement for epithelial-mesenchymal contact and possible role in bronchial smooth muscle development. AB - Laminins, the main components of basement membranes, are heterotrimers consisting of alpha, beta, and gamma polypeptide chains linked together by disulfide bonds. Laminins-1 and -2 are both composed of beta1 and gamma1 chains and differ from each other on their alpha chain, which is alpha1 and alpha2 for laminin-1 and -2, respectively. The present study shows that whereas laminins-1 and -2 are synthesized in the mouse developing lung and in epithelial-mesenchymal cocultures derived from it, epithelial and mesenchymal monocultures lose their ability to synthesize the laminin alpha1 chain. Synthesis of laminin alpha1 chain however returns upon re-establishment of epithelial-mesenchymal contact. Cell-cell contact is critical, since laminin alpha1 chain is not detected in monocultures exposed to coculture-conditioned medium or in epithelial-mesenchymal cocultures in which heterotypic cell-cell contact is prevented by an interposing filter. Immunohistochemical studies on cocultures treated with brefeldin A, an inhibitor of protein secretion, indicated both epithelial and mesenchymal cells synthesize laminin alpha1 chain upon heterotypic cell- cell contact. In a set of functional studies, embryonic lung explants were cultured in the presence of monoclonal antibodies to laminin alpha1, alpha2, and beta/gamma chains. Lung explants exposed to monoclonal antibodies to laminin alpha1 chain exhibited alterations in peribronchial cell shape and decreased smooth muscle development, as indicated by low levels of smooth muscle alpha actin and desmin. Taken together, our studies suggest that laminin alpha1 chain synthesis is regulated by epithelial mesenchymal interaction and may play a role in airway smooth muscle development. PMID- 9334357 TI - Presentation of integrins on leukocyte microvilli: a role for the extracellular domain in determining membrane localization. AB - Adhesion of blood leukocytes to the endothelium involves multiple steps including initial attachment (tethering), rolling, and firm arrest. Presentation of adhesion molecules on leukocyte microvilli can substantially enhance tethering. Localization of L-selectin to microvilli and of CD44 to the planar cell body have been shown to depend upon their transmembrane and cytoplasmic domains. We investigated the role of leukocyte integrin transmembrane and cytoplasmic domains in initiating adhesion under flow and in microvillous localization. Integrins alpha4beta7, alphaLbeta2, and alphaMbeta2 were heterologously expressed in K562 cells. alpha4beta7 initiated adhesion under flow and localized to microvilli, whereas beta2 integrins did not initiate adhesion and localized to the cell body. Chimeric integrins were produced by replacing the alpha4beta7 cytoplasmic and/or transmembrane domains with the homologous domains of alphaLbeta2 or alphaMbeta2. Unexpectedly, these chimeras efficiently mediated adhesion to the alpha4beta7 ligand mucosal addressin cell adhesion molecule-1 under flow and localized to microvilli. Therefore, differences between the transmembrane and cytoplasmic domains of alpha4 and beta2 integrins do not account for differences in ability to support attachment under flow or in membrane localization. Integrins alpha4beta1, alpha5beta1, alpha6Abeta1, alphavbeta3, and alphaEbeta7 also localized to microvilli. Transmembrane proteins known or suspected to associate with extracellular domains of microvillous integrins, including tetraspans and CD47, were concentrated on microvilli as well. These findings suggest that interactions between the extracellular domains of integrins and associated proteins could direct the assembly of multimolecular complexes on leukocyte microvilli. PMID- 9334358 TI - Nitric oxide primes pancreatic beta cells for Fas-mediated destruction in insulin dependent diabetes mellitus. AB - Fas is an apoptosis-inducing surface receptor involved in controlling tissue homeostasis and function at multiple sites. Here we show that beta cells from the pancreata of newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients express Fas and show extensive apoptosis among those cells located in proximity to Fas ligand-expressing T lymphocytes infiltrating the IDDM islets. Normal human pancreatic beta cells that do not constitutively express Fas, become strongly Fas positive after interleuken (IL)-1beta exposure, and are then susceptible to Fas mediated apoptosis. NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthase, prevents IL-1beta-induced Fas expression, whereas the NO donors sodium nitroprusside and nitric oxide releasing compound (NOC)-18, induce functional Fas expression in normal pancreatic beta cells. These findings suggest that NO mediated upregulation of Fas contributes to pancreatic beta cell damage in IDDM. PMID- 9334359 TI - Upregulation of interleukin 8 by oxygen-deprived cells in glioblastoma suggests a role in leukocyte activation, chemotaxis, and angiogenesis. AB - Leukocyte infiltration and necrosis are two biological phenomena associated with the development of neovascularization during the malignant progression of human astrocytoma. Here, we demonstrate expression of interleukin (IL)-8, a cytokine with chemotactic and angiogenic properties, and of IL-8-binding receptors in astrocytoma. IL-8 expression is first observed in low grade astrocytoma in perivascular tumor areas expressing inflammatory cytokines. In glioblastoma, it further localizes to oxygen-deprived cells surrounding necrosis. Hypoxic/anoxic insults on glioblastoma cells in vitro using anaerobic chamber systems or within spheroids developing central necrosis induced an increase in IL-8 messenger RNA (mRNA) and protein expression. mRNA for IL-8-binding chemokine receptors CXCR1, CXCR2, and the Duffy antigen receptor for chemokines (DARC) were found in all astrocytoma grades by reverse transcription/PCR analysis. In situ hybridization and immunohistochemistry localized DARC expression on normal brain and tumor microvascular cells and CXCR1 and CXCR2 expression to infiltrating leukocytes. These results support a model where IL-8 expression is initiated early in astrocytoma development through induction by inflammatory stimuli and later in tumor progression increases due to reduced microenvironmental oxygen pressure. Augmented IL-8 would directly and/or indirectly promote angiogenesis by binding to DARC and by inducing leukocyte infiltration and activation by binding to CXCR1 and CXCR2. PMID- 9334360 TI - Dendritic cells retrovirally transduced with a model antigen gene are therapeutically effective against established pulmonary metastases. AB - Dendritic cells (DCs) are bone marrow-derived leukocytes that function as potent antigen presenting cells capable of initiating T cell-dependent responses from quiescent lymphocytes. DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models. Transduction of DCs with TAA genes may allow stable, prolonged antigen expression as well as the potential for presentation of multiple, or unidentified, epitopes in association with major histocompatibility complex class I and/or class II molecules. To evaluate the potential efficacy of retrovirally transduced DCs, bone marrow cells harvested from BALB/c mice were transduced with either a model antigen gene encoding beta galactosidase (beta-gal) or a control gene encoding rat HER-2/neu (Neu) by coculture with irradiated ecotropic retroviral producer lines. Bone marrow cells were differentiated into DC in vitro using granulocyte/macrophage colony stimulating factor and interleukin-4. After 7 d in culture, cells were 45-78% double positive for DC phenotypic cell surface markers by FACS(R) analysis, and DC transduced with beta-gal were 41-72% positive for beta-gal expression by X-gal staining. In addition, coculture of beta-gal transduced DC with a beta-gal specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal. DC transduced with beta-gal and control rat HER 2/neu were then used to treat 3-d lung metastases in mice bearing an experimental murine tumor CT26.CL25, expressing the model antigen, beta-gal. Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu. In addition, immunization with beta-gal-transduced DCs resulted in the generation of antigen-specific cytotoxic T lymphocytes (CTLs), which were significantly more reactive against relevant tumor targets than CTLs generated from mice immunized with DCs pulsed with the Ld-restricted beta-gal peptide. The results observed in this rapidly lethal tumor model suggest that DCs transduced with TAA may be a useful treatment modality in tumor immunotherapy. PMID- 9334361 TI - Survival of mature CD4 T lymphocytes is dependent on major histocompatibility complex class II-expressing dendritic cells. AB - Thymic T cell development is controlled by T cell receptor (TCR)-major histocompatibility complex (MHC) interactions, whereas a further dependence of peripheral mature T cells on TCR-MHC contact has not been described so far. To study this question, CD4 T cell survival was surveyed in mice lacking MHC class II expression and in mice expressing MHC class II exclusively on dendritic cells. Since neither of these mice positively select CD4 T cells in the thymus, they were grafted with MHC class II-positive embryonic thymic tissue, which had been depleted of bone marrow derived cells. Although the thymus grafts in both hosts were repopulated with host origin thymocytes of identical phenotype and numbers, an accumulation of CD4+ T cells in peripheral lymphoid organs could only be observed in mice expressing MHC class II on dendritic cells, but not in mice that were completely MHC class II deficient. As assessed by histology, the accumulating peripheral CD4 T cells were found to be in close contact with MHC class II+ dendritic cells, suggesting that CD4 T cells need peripheral MHC class II expression for survival and that class II+ dendritic cells might play an important role for the longevity of CD4 T cells. PMID- 9334362 TI - A critical role for lymphotoxin in experimental allergic encephalomyelitis. AB - The lymphotoxin (LT)/tumor necrosis factor (TNF) family has been implicated in the neurologic inflammatory diseases multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE). To determine the role of individual family members in EAE, C57BL/6 mice, LT-alpha-deficient (LT-alpha-/- mice), or LT-beta deficient (LT-beta-/- mice), and their wild-type (WT) littermates were immunized with rat myelin oligodendrocyte glycoprotein (MOG) peptide 35-55. C57BL/6 and WT mice developed chronic, sustained paralytic disease with average maximum clinical scores of 3.5 and disease indices (a measure of day of onset and sustained disease scores) ranging from 367 to 663 with central nervous system (CNS) inflammation and demyelination. LT-alpha-/- mice were primed so that their splenic lymphocytes proliferated in response to MOG 35-55 and the mice produced anti-MOG antibody. However, LT-alpha-/- mice were quite resistant to EAE with low average clinical scores (<1), an average disease index of 61, and the negligible CNS inflammation and demyelination. WT T cells transferred EAE to LT-alpha-/- recipients. LT-beta-/- mice were susceptible to EAE, though less than WT, with an average maximum clinical score of 1.9 and disease index of 312. These data implicate T cell production of LT-alpha in MOG EAE and support a major role for LT-alpha3, a minor role for the LT-alpha/beta complex, and by inference, no role for TNF-alpha. PMID- 9334363 TI - Mycobacterium tuberculosis chaperonin 10 stimulates bone resorption: a potential contributory factor in Pott's disease. AB - Pott's disease (spinal tuberculosis), a condition characterized by massive resorption of the spinal vertebrae, is one of the most striking pathologies resulting from local infection with Mycobacterium tuberculosis (Mt; Boachie Adjei, O., and R.G. Squillante. 1996. Orthop. Clin. North Am. 27:95-103). The pathogenesis of Pott's disease is not established. Here we report for the first time that a protein, identified by a monoclonal antibody to be the Mt heat shock protein (Baird, P.N., L.M. Hall, and A.R.M. Coates. 1989. J. Gen. Microbiol. 135:931-939) chaperonin (cpn) 10, is responsible for the osteolytic activity of this bacterium. Recombinant Mt cpn10 is a potent stimulator of bone resorption in bone explant cultures and induces osteoclast recruitment, while inhibiting the proliferation of an osteoblast bone-forming cell line. Furthermore, we have found that synthetic peptides corresponding to sequences within the flexible loop and sequence 65-70 of Mt cpn10 may comprise a single conformational unit which encompasses its potent bone-resorbing activity. Our findings suggest that Mt cpn10 may be a valuable pharmacological target for the clinical therapy of vertebral tuberculosis and possibly other bone diseases. PMID- 9334364 TI - Dendritic cells genetically modified with an adenovirus vector encoding the cDNA for a model antigen induce protective and therapeutic antitumor immunity. AB - Dendritic cells (DCs) are potent antigen-presenting cells that play a critical role in the initiation of antitumor immune responses. In this study, we show that genetic modifications of a murine epidermis-derived DC line and primary bone marrow-derived DCs to express a model antigen beta-galactosidase (betagal) can be achieved through the use of a replication-deficient, recombinant adenovirus vector, and that the modified DCs are capable of eliciting antigen-specific, MHC restricted CTL responses. Importantly, using a murine metastatic lung tumor model with syngeneic colon carcinoma cells expressing betagal, we show that immunization of mice with the genetically modified DC line or bone marrow DCs confers potent protection against a lethal tumor challenge, as well as suppression of preestablished tumors, resulting in a significant survival advantage. We conclude that genetic modification of DCs to express antigens that are also expressed in tumors can lead to antigen-specific, antitumor killer cells, with a concomitant resistance to tumor challenge and a decrease in the size of existing tumors. PMID- 9334365 TI - Regulation of anti-double-stranded DNA B cells in nonautoimmune mice: localization to the T-B interface of the splenic follicle. AB - Systemic lupus erythematosus (SLE) and the MRL-lpr/lpr murine model for SLE are characterized by the presence of serum anti-double-stranded (ds)DNA antibodies (Abs), whereas nonautoimmune individuals have negligible levels of these Abs. To increase the frequency of anti-DNA B cells and identify the mechanisms involved in their regulation in nonautoimmune mice, we have used Ig transgenes (tgs). In the present study, we used the VH3H9 heavy (H) chain tg which expresses an H chain that was repeatedly isolated from anti-dsDNA Abs from MRL-lpr/lpr mice. Because the VH3H9 H chain can pair with endogenous L chains to generate anti single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire. We have identified anti-dsDNA B cells that are located at the T-B interface in the splenic follicle where they have an increased in vivo turnover rate. These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure. PMID- 9334366 TI - Peripheral T cell survival requires continual ligation of the T cell receptor to major histocompatibility complex-encoded molecules. AB - In the thymus, T cells are selected according to their T cell receptor (TCR) specificity. After positive selection, mature cells are exported from primary lymphoid organs to seed the secondary lymphoid tissue. An important question is whether survival of mature T cells is an intrinsic property or requires continuous survival signals, i.e., engagement of the TCR by major histocompatibility complex (MHC) molecules in the periphery, perhaps in a similar way as occurring during thymic positive selection. To address this issue we used recombination-activating gene (Rag)-deficient H-2b mice expressing a transgenic TCR restricted by I-Ed class II MHC molecules. After engraftment with Rag-/- H-2d fetal thymi, CD4+8- peripheral T cells emerged. These cells were isolated and transferred into immunodeficient hosts of H-2b or H-2d haplotype, some of the latter being common cytokine receptor gamma chain deficient to exclude rejection of H-2b donor cells by host natural killer cells. Our results show that in the absence, but not in the presence, of selecting MHC molecules, peripheral mature T cells are short lived and disappear within 7 wk, indicating that continuous contact of the TCR with selecting MHC molecules is required for survival of T cells. PMID- 9334367 TI - The common cytokine receptor gamma chain controls survival of gamma/delta T cells. AB - We have investigated the role of common gamma chain (gamma c)-signaling pathways for the development of T cell receptor for antigen (TCR)-gamma/delta T cells. TCR gamma/delta-bearing cells were absent from the adult thymus, spleen, and skin of gamma c-deficient (gamma c-) mice, whereas small numbers of thymocytes expressing low levels of TCR-gamma/delta were detected during fetal life. Recent reports have suggested that signaling via interleukin (IL)-7 plays a major role in facilitating TCR-gamma/delta development through induction of V-J (variable joining) rearrangements at the TCR-gamma locus. In contrast, we detected clearly TCR-gamma rearrangements in fetal thymi from gamma c- mice (which fail to signal in response to IL-7) and reduced TCR-gamma rearrangements in adult gamma c thymi. No gross defects in TCR-delta or TCR-beta rearrangements were observed in gamma c mice of any age. Introduction of productively rearranged TCR V gamma 1 or TCR V gamma 1/V delta 6 transgenes onto mice bearing the gamma c mutation did not restore TCR-gamma/delta development to normal levels suggesting that gamma c dependent pathways provide additional signals to developing gamma/delta T cells other than for the recombination process. Bcl-2 levels in transgenic thymocytes from gamma c- mice were dramatically reduced compared to gamma c+ transgenic littermates. We favor the concept that gamma c-dependent receptors are required for the maintenance of TCR-gamma/delta cells and contribute to the completion of TCR-gamma rearrangements primarily by promoting survival of cells committed to the TCR-gamma/delta lineage. PMID- 9334368 TI - Inhibition of virus attachment to CD4+ target cells is a major mechanism of T cell line-adapted HIV-1 neutralization. AB - Antibody-mediated neutralization of human immunodeficiency virus type-1 (HIV-1) is thought to function by at least two distinct mechanisms: inhibition of virus receptor binding, and interference with events after binding, such as virus-cell membrane fusion. Here we show, by the use of a novel virus-cell binding assay, that soluble CD4 and monoclonal antibodies to all confirmed glycoprotein (gp)120 neutralizing epitopes, including the CD4 binding site and the V2 and V3 loops, inhibit the adsorption of two T cell line-adapted HIV-1 viruses to CD4+ cells. A correlation between the inhibition of virus binding and virus neutralization was observed for soluble CD4 and all anti-gp120 antibodies, indicating that this is a major mechanism of HIV neutralization. By contrast, antibodies specific for regions of gp120 other than the CD4 binding site showed little or no inhibition of either soluble gp120 binding to CD4+ cells or soluble CD4 binding to HIV infected cells, implying that this effect is specific to the virion-cell interaction. However, inhibition of HIV-1 attachment to cells is not a universal mechanism of neutralization, since an anti-gp41 antibody did not inhibit virus cell binding at neutralizing concentrations, implying activity after virus-cell binding. PMID- 9334369 TI - Delivery of B cell receptor-internalized antigen to endosomes and class II vesicles. AB - B cell receptor (BCR)-mediated antigen processing is a mechanism that allows class II-restricted presentation of specific antigen by B cells at relatively low antigen concentrations. Although BCR-mediated antigen processing and class II peptide loading may occur within one or more endocytic compartments, the functions of these compartments and their relationships to endosomes and lysosomes remain uncertain. In murine B cells, at least one population of class II- containing endocytic vesicles (i.e., CIIV) has been identified and demonstrated to be distinct both physically and functionally from endosomes and lysosomes. We now demonstrate the delivery of BCR-internalized antigen to CIIV within the time frame during which BCR-mediated antigen processing and formation of peptide-class II complexes occurs. Only a fraction of the BCR-internalized antigen was delivered to CIIV, with the majority of internalized antigen being delivered to lysosomes that are largely class II negative. The extensive colocalization of BCR-internalized antigen and newly synthesized class II molecules in CIIV suggests that CIIV may represent a specialized subcellular compartment for BCR-mediated antigen processing. Additionally, we have identified a putative CIIV-marker protein, immunologically related to the Igalpha subunit of the BCR, which further illustrates the unique nature of these endocytic vesicles. PMID- 9334370 TI - Negative regulation of Fc epsilon RI-mediated degranulation by CD81. AB - Signaling through the high affinity receptor for immunoglobulin E (Fc epsilon RI) results in the coordinate activation of tyrosine kinases before calcium mobilization. Receptors capable of interfering with the signaling of antigen receptors, such as Fc epsilon RI, recruit tyrosine and inositol phosphatases that results in diminished calcium mobilization. Here, we show that antibodies recognizing CD81 inhibit Fc epsilon RI-mediated mast cell degranulation but, surprisingly, without affecting aggregation-dependent tyrosine phosphorylation, calcium mobilization, or leukotriene synthesis. Furthermore, CD81 antibodies also inhibit mast cell degranulation in vivo as measured by reduced passive cutaneous anaphylaxis responses. These results reveal an unsuspected calcium-independent pathway of antigen receptor regulation, which is accessible to engagement by membrane proteins and on which novel therapeutic approaches to allergic diseases could be based. PMID- 9334371 TI - Heat shock protein-peptide complexes, reconstituted in vitro, elicit peptide specific cytotoxic T lymphocyte response and tumor immunity. AB - Heat shock protein (HSP) preparations derived from cancer cells and virus infected cells have been shown previously to elicit cancer-specific or virus specific immunity. The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs. The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro. The studies show that (a) complexes of hsp70 or gp96 HSP molecules with a variety of synthetic peptides can be generated in vitro; (b) the binding of HSPs with peptides is specific in that a number of other proteins tested do not bind synthetic peptides under the conditions in which gp96 molecules do; (c) HSP peptide complexes reconstituted in vitro are immunologically active, as tested by their ability to elicit antitumor immunity and specific CD8+ cytolytic T lymphocyte response; and (d) synthetic peptides reconstituted in vitro with gp96 are capable of being taken up and re-presented by macrophage in the same manner as gp96- peptides complexes generated in vivo. These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants. PMID- 9334372 TI - Granule-mediated killing: pathways for granzyme B-initiated apoptosis. AB - We report that the serine protease granzyme B (GrB), which is crucial for granule mediated cell killing, initiates apoptosis in target cells by first maturing caspase-10. In addition, GrB has a limited capacity to mature other caspases and to cause cell death independently of the caspases. Compared with other members, GrB in vitro most efficiently processes caspase-7 and -10. In a human cell model, full maturation of caspase-7 does not occur unless caspase-10 is present. Furthermore, GrB matured caspase-3 with less efficiency than caspase-7 or caspase 10. With the caspases fully inactivated by peptidic inhibitors, GrB induced in Jurkat cells growth arrest and, over a delayed time period, cell death. Thus, the primary mechanism by which GrB initiates cell death is activation of the caspases through caspase-10. However, under circumstances where caspase-10 is absent or dysfunctional, GrB can act through secondary mechanisms including activation of other caspases and direct cell killing by cleavage of noncaspase substrates. The redundant functions of GrB ensure the effectiveness of granule-mediated cell killing, even in target cells that lack the expression or function (e.g., by mutation or a viral serpin) of one or more of the caspases, providing the host with overlapping safeguards against aberrantly replicating, nonself or virally infected cells. PMID- 9334373 TI - Analysis of the expression of peptide-major histocompatibility complexes using high affinity soluble divalent T cell receptors. AB - Understanding the regulation of cell surface expression of specific peptide-major histocompatibility complex (MHC) complexes is hindered by the lack of direct quantitative analyses of specific peptide-MHC complexes. We have developed a direct quantitative biochemical approach by engineering soluble divalent T cell receptor analogues (TCR-Ig) that have high affinity for their cognate peptide-MHC ligands. The generality of this approach was demonstrated by specific staining of peptide-pulsed cells with two different TCR-Ig complexes: one specific for the murine alloantigen 2C, and one specific for a viral peptide from human T lymphocyte virus-1 presented by human histocompatibility leukocyte antigens-A2. Further, using 2C TCR- Ig, a more detailed analysis of the interaction with cognate peptide-MHC complexes revealed several interesting findings. Soluble divalent 2C TCR-Ig detected significant changes in the level of specific antigenic-peptide MHC cell surface expression in cells treated with gamma interferon (gamma-IFN). Interestingly, the effects of gamma-IFN on expression of specific peptide-MHC complexes recognized by 2C TCR-Ig were distinct from its effects on total H-2 Ld expression; thus, lower doses of gamma-IFN were required to increase expression of cell surface class I MHC complexes than were required for upregulation of expression of specific peptide-MHC complexes. Analysis of the binding of 2C TCR-Ig for specific peptide-MHC ligands unexpectedly revealed that the affinity of the 2C TCR-Ig for the naturally occurring alloreactive, putatively, negatively selecting, complex, dEV-8-H-2 Kbm3, is very low, weaker than 71 microM. The affinity of the 2C TCR for the other naturally occurring, negatively selecting, alloreactive complex, p2Ca-H-2 Ld, is approximately 1000 fold higher. Thus, negatively selecting peptide-MHC complexes do not necessarily have intrinsically high affinity for cognate TCR. These results, uniquely revealed by this analysis, indicate the importance of using high affinity biologically relevant cognates, such as soluble divalent TCR, in furthering our understanding of immune responses. PMID- 9334374 TI - Mutational evidence for control of cell adhesion through integrin diffusion/clustering, independent of ligand binding. AB - Previous studies have shown that integrin alpha chain tails make strong positive contributions to integrin-mediated cell adhesion. We now show here that integrin alpha4 tail deletion markedly impairs static cell adhesion by a mechanism that does not involve altered binding of soluble vascular cell adhesion molecule 1 ligand. Instead, truncation of the alpha4 cytoplasmic domain caused a severe deficiency in integrin accumulation into cell surface clusters, as induced by ligand and/ or antibodies. Furthermore, alpha4 tail deletion also significantly decreased the membrane diffusivity of alpha4beta1, as determined by a single particle tracking technique. Notably, low doses of cytochalasin D partially restored the deficiency in cell adhesion seen upon alpha4 tail deletion. Together, these results suggest that alpha4 tail deletion exposes the beta1 cytoplasmic domain, leading to cytoskeletal associations that apparently restrict integrin lateral diffusion and accumulation into clusters, thus causing reduced static cell adhesion. Our demonstration of integrin adhesive activity regulated through receptor diffusion/clustering (rather than through altered ligand binding affinity) may be highly relevant towards the understanding of inside-out signaling mechanisms for beta1 integrins. PMID- 9334375 TI - Neutrophil emigration in the skin, lungs, and peritoneum: different requirements for CD11/CD18 revealed by CD18-deficient mice. AB - To determine the role of CD11/CD18 complexes in neutrophil emigration, inflammation was induced in the skin, lungs, or peritoneum of mutant mice deficient in CD18 (CD18-/- mutants). Peripheral blood of CD18-/- mutants contained 11-fold more neutrophils than did blood of wild-type (WT) mice. During irritant dermatitis induced by topical application of croton oil, the number of emigrated neutrophils in histological sections of dermis was 98% less in CD18-/- mutants than in WT mice. During Streptococcus pneumoniae pneumonia, neutrophil emigration in CD18-/- mutants was not reduced. These data are consistent with expectations based on studies using blocking antibodies to inhibit CD11/CD18 complexes, and on observations of humans lacking CD11/CD18 complexes. The number of emigrated neutrophils in lung sections during Escherichia coli pneumonia, or in peritoneal lavage fluid after 4 h of S. pneumoniae peritonitis, was not reduced in CD18-/- mutants, but rather was greater than the WT values (240 +/- 30 and 220 +/- 30% WT, respectively). Also, there was no inhibition of neutrophil emigration during sterile peritonitis induced by intraperitoneal injection of thioglycollate (90 +/- 20% WT). These data contrast with expectations. Whereas CD11/CD18 complexes are essential to the dermal emigration of neutrophils during acute dermatitis, CD18-/- mutant mice demonstrate surprising alternative pathways for neutrophil emigration during pneumonia or peritonitis. PMID- 9334376 TI - Interleukin-1 beta converting enzyme-like protease involvement in Fas-induced and activation-induced peripheral blood T cell apoptosis in HIV infection. TNF related apoptosis-inducing ligand can mediate activation-induced T cell death in HIV infection. AB - Apoptosis of peripheral blood T cells has been suggested to play an important role in the pathogenesis of human immunodeficiency virus (HIV) infection. Spontaneous, Fas (CD95)-induced and activation-induced T cell apoptosis have all been described in peripheral blood mononuclear cell cultures of HIV-infected individuals. We have previously shown that activation-induced T cell apoptosis is Fas independent in peripheral blood T cells from HIV+ individuals. In this study, we extend and confirm these observations by using an inhibitor of interleukin-1 beta converting enzyme (ICE) homologues. We show that z-VAD-fmk, a tripeptide inhibitor of ICE homologues, can inhibit Fas-induced apoptosis of peripheral blood CD4(+) and CD8+ T cells from asymptomatic HIV+ individuals. z-VAD-fmk also inhibited activation (anti-CD3)- induced CD4+ and CD8+ T cell apoptosis (AICD) in some but not all asymptomatic HIV+ individuals. Apoptosis was measured by multiparameter flow cytometry. The z-VAD-fmk inhibitor also enhanced survival of T cells in anti-Fas or anti-CD3 antibody-treated cultures and inhibited DNA fragmentation. AICD that could be inhibited by z-VAD-fmk was Fas independent and could be inhibited with a blocking monoclonal antibody to tumor necrosis factor related apoptosis-inducing ligand (TRAIL), a recently described member of the TNF/nerve growth factor ligand family. The above findings show that Fas-induced T cell apoptosis is ICE dependent in HIV infection. AICD can be blocked by ICE inhibitors in some patients, and this AICD is mediated by TRAIL. These results show that TRAIL can be a mediator of AICD in T cells. These different mechanisms of peripheral blood T cell apoptosis may play different roles in the pathogenesis of HIV infection. PMID- 9334377 TI - Interaction of chemokine receptor CCR5 with its ligands: multiple domains for HIV 1 gp120 binding and a single domain for chemokine binding. AB - CCR5 is a chemokine receptor expressed by T cells and macrophages, which also functions as the principal coreceptor for macrophage (M)-tropic strains of HIV-1. To understand the molecular basis of the binding of chemokines and HIV-1 to CCR5, we developed a number of mAbs that inhibit the various interactions of CCR5, and mapped the binding sites of these mAbs using a panel of CCR5/CCR2b chimeras. One mAb termed 2D7 completely blocked the binding and chemotaxis of the three natural chemokine ligands of CCR5, RANTES (regulated on activation normal T cell expressed and secreted), macrophage inflammatory protein (MIP)-1alpha, and MIP 1beta, to CCR5 transfectants. This mAb was a genuine antagonist of CCR5, since it failed to stimulate an increase in intracellular calcium concentration in the CCR5 transfectants, but blocked calcium responses elicited by RANTES, MIP-1alpha, or MIP-1beta. This mAb inhibited most of the RANTES and MIP-1alpha chemotactic responses of activated T cells, but not of monocytes, suggesting differential usage of chemokine receptors by these two cell types. The 2D7 binding site mapped to the second extracellular loop of CCR5, whereas a group of mAbs that failed to block chemokine binding all mapped to the NH2-terminal region of CCR5. Efficient inhibition of an M-tropic HIV-1-derived envelope glycoprotein gp120 binding to CCR5 could be achieved with mAbs recognizing either the second extracellular loop or the NH2-terminal region, although the former showed superior inhibition. Additionally, 2D7 efficiently blocked the infectivity of several M-tropic and dual-tropic HIV-1 strains in vitro. These results suggest a complicated pattern of HIV-1 gp120 binding to different regions of CCR5, but a relatively simple pattern for chemokine binding. We conclude that the second extracellular loop of CCR5 is an ideal target site for the development of inhibitors of either chemokine or HIV-1 binding to CCR5. PMID- 9334378 TI - Inhibition of T-tropic HIV strains by selective antagonization of the chemokine receptor CXCR4. AB - Bicyclams are a novel class of antiviral compounds that are highly potent and selective inhibitors of the replication of HIV-1 and HIV-2. Surprisingly, however, when the prototype compound AMD3100 was tested against M-tropic virus strains such as BaL, ADA, JR-CSF, and SF-162 in human peripheral blood mononuclear cells, the compound was completely inactive. Because of the specific and potent inhibitory effect of AMD3100 on T-tropic viruses, but not M-tropic viruses, it was verified that AMD3100 interacts with the CXC-chemokine receptor CXCR4, the main coreceptor used by T-tropic viruses. AMD3100 dose dependently inhibited the binding of a specific CXCR4 monoclonal antibody to SUP-T1 cells as measured by flow cytometry. It did not inhibit the binding of the biotinylated CC chemokine macrophage inflammatory protein (MIP) 1alpha or MIP-1beta, ligands for the chemokine receptor CCR5 (the main coreceptor for M-tropic viruses). In addition, AMD3100 completely blocked (a) the Ca2+ flux at 100 ng/ml in lymphocytic SUP-T1 and monocytic THP-1 cells, and (b) the chemotactic responses of THP-1 cells induced by stromal cell-derived factor 1alpha, the natural ligand for CXCR4. Finally, AMD3100 had no effect on the Ca2+ flux induced by the CC chemokines MIP-1alpha, regulated on activation normal T cell expressed and secreted (RANTES; also a ligand for CCR5), or monocyte chemoattractant protein 3 (a ligand for CCR1 and CCR2b), nor was it able to induce Ca2+ fluxes by itself. The bicyclams are, to our knowledge, the first low molecular weight anti-HIV agents shown to act as potent and selective CXCR4 antagonists. PMID- 9334379 TI - A small molecule CXCR4 inhibitor that blocks T cell line-tropic HIV-1 infection. AB - Several members of the chemokine receptor family have been shown to function in association with CD4 to permit human immunodeficiency virus type 1 (HIV-1) entry and infection. The CXC chemokine receptor CXCR4/fusin is a receptor for pre-B cell growth stimulating factor (PBSF)/stromal cell-derived factor 1 (SDF-1) and serves as a coreceptor for the entry of T cell line-tropic HIV-1 strains. Thus, the development of CXCR4 antagonists or agonists may be useful in the treatment of HIV-1 infection. T22 ([Tyr5,12,Lys7]-polyphemusin II) is a synthesized peptide that consists of 18 amino acid residues and an analogue of polyphemusin II isolated from the hemocyte debris of American horseshoe crabs (Limulus polyphemus). T22 was found to specifically inhibit the ability of T cell line tropic HIV-1 to induce cell fusion and infect the cell lines transfected with CXCR4 and CD4 or peripheral blood mononuclear cells. In addition, T22 inhibited Ca2+ mobilization induced by pre-B cell growth stimulating factor (PBSF)/SDF-1 stimulation through CXCR4. Thus, T22 is a small molecule CXCR4 inhibitor that blocks T cell line-tropic HIV-1 entry into target cells. PMID- 9334381 TI - Suppression of leukotriene B4 biosynthesis by endogenous adenosine in ligand activated human neutrophils. AB - Adenosine (Ado) has been shown to suppress several functional responses of human polymorphonuclear leukocytes (PMNs). The current study investigated whether endogenous Ado regulates the biosynthesis of leukotriene (LT)B4 in ligand stimulated PMNs. Measurements of Ado in PMN resuspended in Hanks' buffered salt solution (HBSS) or plasma showed a cell concentration- and time-dependent accumulation of the nucleoside. The removal of endogenous Ado with either Ado deaminase or the blockade of its action by the Ado A2a receptor antagonist, 8-(3 chlorostyryl) caffeine, markedly increased LTB4 biosynthesis upon ligand stimulation in HBSS. Similarly, LTB4 synthesis by ligand-stimulated PMNs in plasma (containing recombinant LTA4 hydrolase to allow the conversion of protein bound LTA4) was strongly enhanced by addition of Ado deaminase. Addition of red blood cells to suspensions of PMNs in plasma mimicked the effect of adding Ado deaminase and LTA4 hydrolase in enhancing LTB4 biosynthesis upon ligand stimulation. This effect of red blood cells on LTB4 biosynthesis was blocked by dipyridamole, an inhibitor of Ado transport, or captopril, an inhibitor of LTA4 hydrolase. These results demonstrate that endogenous Ado efficiently downregulates ligand-stimulated LTB4 biosynthesis in PMN suspensions, pointing out a potentially important regulatory function of Ado in inflammatory exudates. These results also unveil a dual role for red blood cells in upregulating LTB4 biosynthesis, namely, the removal of endogenous Ado and the conversion of LTA4 released by activated PMNs. PMID- 9334380 TI - A small-molecule inhibitor directed against the chemokine receptor CXCR4 prevents its use as an HIV-1 coreceptor. AB - The chemokine receptor CXCR4 is the major coreceptor used for cellular entry by T cell- tropic human immunodeficiency virus (HIV)-1 strains, whereas CCR5 is used by macrophage (M)-tropic strains. Here we show that a small-molecule inhibitor, ALX40-4C, inhibits HIV-1 envelope (Env)-mediated membrane fusion and viral entry directly at the level of coreceptor use. ALX40-4C inhibited HIV-1 use of the coreceptor CXCR4 by T- and dual-tropic HIV-1 strains, whereas use of CCR5 by M- and dual-tropic strains was not inhibited. Dual-tropic viruses capable of using both CXCR4 and CCR5 were inhibited by ALX40-4C only when cells expressed CXCR4 alone. ALX40-4C blocked stromal-derived factor (SDF)-1alpha-mediated activation of CXCR4 and binding of the monoclonal antibody 12G5 to cells expressing CXCR4. Overlap of the ALX40-4C binding site with that of 12G5 and SDF implicates direct blocking of Env interactions, rather than downregulation of receptor, as the mechanism of inhibition. Thus, ALX40-4C represents a small-molecule inhibitor of HIV-1 infection that acts directly against a chemokine receptor at the level of Env-mediated membrane fusion. PMID- 9334382 TI - Caenorhabditis elegans rab-3 mutant synapses exhibit impaired function and are partially depleted of vesicles. AB - Rab molecules regulate vesicular trafficking in many different exocytic and endocytic transport pathways in eukaryotic cells. In neurons, rab3 has been proposed to play a crucial role in regulating synaptic vesicle release. To elucidate the role of rab3 in synaptic transmission, we isolated and characterized Caenorhabditis elegans rab-3 mutants. Similar to the mouse rab3A mutants, these mutants survived and exhibited only mild behavioral abnormalities. In contrast to the mouse mutants, synaptic transmission was perturbed in these animals. Extracellular electrophysiological recordings revealed that synaptic transmission in the pharyngeal nervous system was impaired. Furthermore, rab-3 animals were resistant to the acetylcholinesterase inhibitor aldicarb, suggesting that cholinergic transmission was generally depressed. Last, synaptic vesicle populations were redistributed in rab-3 mutants. In motor neurons, vesicle populations at synapses were depleted to 40% of normal levels, whereas in intersynaptic regions of the axon, vesicle populations were elevated. On the basis of the morphological defects at neuromuscular junctions, we postulate that RAB-3 may regulate recruitment of vesicles to the active zone or sequestration of vesicles near release sites. PMID- 9334383 TI - Developmental expression pattern of phototransduction components in mammalian pineal implies a light-sensing function. AB - Whereas the pineal organs of lower vertebrates have been shown to be photosensitive, photic regulation of pineal function in adult mammals is thought be mediated entirely by retinal photoreceptors. Extraretinal regulation of pineal function has been reported in neonatal rodents, although both the site and molecular basis of extraretinal photoreception have remained obscure. In this study we examine the developmental expression pattern of all of the principal components of retinal phototransduction in rat pineal via cRNA in situ hybridization. All of the components needed to reconstitute a functional phototransduction pathway are expressed in the majority of neonatal pinealocytes, although the expression levels of many of these genes decline dramatically during development. These findings strongly support the theory that the neonatal rat pineal itself is photosensitive. In addition, we observe in neonatal pinealocytes the expression of both rod-specific and cone-specific phototransduction components, implying the existence of functionally different subtypes of pinealocytes that express varying combinations of phototransduction enzymes. PMID- 9334384 TI - Parapinopsin, a novel catfish opsin localized to the parapineal organ, defines a new gene family. AB - Multiple sites of extraretinal photoreception are present in vertebrates, but the molecular basis of extraretinal phototransduction is poorly understood. This study reports the cloning of the first opsin specifically expressed in the directly photosensitive pineal and parapineal of cold-blooded vertebrates. This opsin, identified in channel catfish and termed parapinopsin, defines a new gene family of vertebrate photopigments and is expressed in a majority of parapinealocytes and a subset of pineal photoreceptor cells. Parapinopsin shows a caudal-rostral gradient of expression within the pineal organ. This study also reports the cloning of partial cDNAs encoding the channel catfish orthologues of rhodopsin and the red cone pigment-the full complement of retinal opsins in the species. In situ hybridization studies using probes derived from these retinal opsins, together with parapinopsin, reveal no expression of retinal opsins in pineal and parapineal organ and no expression of any opsin tested in the "deep brain," iris, or dermal melanophores. These data imply that phototransduction in these sites of extraretinal photoreception must be mediated by novel opsins. PMID- 9334385 TI - Contingent-dependent enhancement of rhythmic motor patterns: an in vitro analog of operant conditioning. AB - Operant conditioning is characterized by the contingent reinforcement of a designated behavior. Previously, feeding behavior in Aplysia has been demonstrated to be modified by operant conditioning, and a neural pathway (esophageal nerve; E n.) that mediates some aspects of reinforcement has been identified. As a first step toward a cellular analysis of operant conditioning, we developed an in vitro buccal ganglia preparation that expressed the essential features of operant conditioning. Motor patterns that represented at least two different aspects of fictive feeding (i.e., ingestion-like and rejection-like motor patterns) were elicited by tonic stimulation of a peripheral buccal nerve (n.2,3). Three groups of preparations were examined. In a contingent reinforcement group, stimulation of E n. was contingent on the expression of a specific type of motor pattern (i.e., either ingestion-like or rejection-like). In a yoke-control group, stimulation of E n. was not contingent on any specific pattern. In a control group, E n. was not stimulated. The frequency of the reinforced pattern increased significantly only in the contingent-reinforcement group. No changes were observed in nonreinforced patterns or in the motor patterns of the control and yoke-control groups. Contingent reinforcement of the ingestion-like pattern was associated with an enhancement of activity in motor neuron B8, and this enhancement was specific to the reinforced pattern. These results suggest that the isolated buccal ganglia expressed an essential feature of operant conditioning (i.e., contingent reinforcement modified a designated operant) and that this analog of operant conditioning is accessible to cellular analysis. PMID- 9334386 TI - Instantaneous perturbation of dentate interneuronal networks by a pressure wave transient delivered to the neocortex. AB - Whole-cell patch-clamp recordings and immunocytochemical experiments were performed to determine the short- and long-term effects of lateral fluid percussion head injury on the perisomatic inhibitory control of dentate granule cells in the adult rat, with special reference to the development of trauma induced hyperexcitability. One week after the delivery of a single, moderate (2.0 2.2 atm) mechanical pressure wave to the neocortex, the feed-forward inhibitory control of dentate granule cell discharges was compromised, and the frequency of miniature IPSCs was decreased. Consistent with the electrophysiological data, the number of hilar parvalbumin (PV)- and cholecystokinin (CCK)-positive dentate interneurons supplying the inhibitory innervation of the perisomatic region of granule cells was decreased weeks and months after head injury. The initial injury to the hilar neurons took place instantaneously after the impact and did not require the recruitment of active physiological processes. Furthermore, the decrease in the number of PV- and CCK-positive hilar interneurons was similar to the decrease in the number of the AMPA-type glutamate receptor subunit 2/3 immunoreactive mossy cells, indicating that the pressure wave-transient causes injurious physical stretching and bending of most cells that are large and not tightly packed in a cell layer. These results reveal for the first time that moderate pressure wave-transients, triggered by traumatic head injury episodes, impact the dentate neuronal network in a unique temporal and spatial pattern, resulting in a net decrease in the perisomatic control of granule cell discharges. PMID- 9334387 TI - The effect of peripherin/rds haploinsufficiency on rod and cone photoreceptors. AB - Haploinsufficiency because of a null mutation in the gene encoding peripherin/rds has been thought to be the primary defect associated with the photoreceptor degeneration seen in the retinal degeneration slow (rds) mouse. We have compared the effects of this haploinsufficiency on rod and cone photoreceptors by measuring the levels of rod- and cone-specific gene expression, by determining the relative rates of rod and cone degeneration, and by electroretinography. These analyses were performed at ages before and after the onset of degeneration of the photoreceptor cells. The data were consistent in demonstrating that measures for cone photoreceptors are relatively spared in comparison to comparable measures for rod photoreceptors. Blue cones were retained in higher number than red/green cones for the first 3 months of the degeneration. Our results indicate that the haploinsufficiency present in rds/+ mice has a greater impact on the rod than on the cone photoreceptor, a finding that likely reflects the tight regulation of peripherin/rds and the need for two functional alleles to assemble the structure of the rod outer segment and/or differences between the ultrastructure of the rod and cone outer segments. PMID- 9334388 TI - Neurokinin 1 receptor internalization in spinal cord slices induced by dorsal root stimulation is mediated by NMDA receptors. AB - The excitability of spinal neurons that transmit pain is modulated by glutamate and substance P (SP). Glutamate is an excitatory neurotransmitter in the dorsal horn, and its effects are enhanced by SP acting on neurokinin 1 receptors (NK1Rs). We assessed activation of NK1Rs by studying their internalization in spinal cord slices. NK1Rs were localized in sections from the slices by using immunohistochemistry combined with fluorescence and confocal microscopy. Incubating the slices with SP induced internalization in most NK1R-positive neurons in laminae I, IIo, and X and in half of NK1R-positive neurons in laminae III-V. SP-induced internalization was abolished by the specific NK1R antagonist L 703,606 (1 microM). Stimulating the dorsal root with long-duration (0.4 msec) pulses evoked EPSPs in dorsal horn neurons with latencies consistent with the conduction speed of A partial differential- and C-fibers. High-frequency (100 Hz) stimulation of the dorsal root with these pulses induced NK1R internalization in neurons in laminae I-IIo of the stimulated side of the slice but not in the contralateral side or in other laminae. Stimulation at lower frequencies (1 and 10 Hz) failed to elicit significant internalization, suggesting that the release of SP is frequency-dependent. Internalization produced by the 100 Hz tetanus was mimicked by NMDA and blocked by an NMDA antagonist, 2-amino-5-phosphonopentanoic acid, but not by the AMPA and kainate antagonist CNQX. The NK1R antagonist L 703,606 abolished the internalization produced by 100 Hz stimulation or NMDA. Therefore, the release of SP in the dorsal horn appears to be controlled by NMDA receptors. PMID- 9334389 TI - Presynaptic depression at a calyx synapse: the small contribution of metabotropic glutamate receptors. AB - Synaptic depression of evoked EPSCs was quantified with stimulation frequencies ranging from 0.2 to 100 Hz at the single CNS synapse formed by the calyx of Held in the rat brainstem. Half-maximal depression occurred at approximately 1 Hz, with 10 and 100 Hz stimulation frequencies reducing EPSC amplitudes to approximately 30% and approximately 10% of their initial magnitude, respectively. The time constant of recovery from depression elicited by 10 Hz afferent fiber stimulation was 4.2 sec. AMPA and NMDA receptor-mediated EPSCs depressed in parallel at 1-5 Hz stimulation frequencies, suggesting that depression was induced by presynaptic mechanism(s) that reduced glutamate release. To determine the contribution of autoreceptors to depression, we studied the inhibitory effects of the metabotropic glutamate receptor (mGluR) agonists (1S, 3S)-ACPD and L-AP4 and found them to be reversed in a dose-dependent manner by (RS)-alpha cyclopropyl-4-phosphonophenylglycine (CPPG), a novel and potent competitive antagonist of mGluRs. At 300 microM, CPPG completely reversed the effects of L AP4 and (1S, 3S)-ACPD, but reduced 5-10 Hz elicited depression by only approximately 6%. CPPG-sensitive mGluRs, presumably activated by glutamate spillover during physiological synaptic transmission, thus contribute on the order of only 10% to short-term synaptic depression. We therefore suggest that the main mechanism contributing to the robust depression elicited by 5-10 Hz afferent fiber stimulation of the calyx of Held synapse is synaptic vesicle pool depletion. PMID- 9334390 TI - Bidirectional regulation of DARPP-32 phosphorylation by dopamine. AB - Dopamine has been shown to stimulate phosphorylation of DARPP-32, a phosphoprotein highly enriched in medium-sized spiny neurons of the neostriatum. Here, we investigated the contribution of D1-like and D2-like dopamine receptors in the regulation of DARPP-32 phosphorylation in mouse striatal slices. D1-like and D2-like receptors had opposing effects on the state of DARPP-32 phosphorylation. The D1 receptor agonist SKF82526 increased DARPP-32 phosphorylation. In contrast, the D2 receptor agonist quinpirole decreased basal as well as D1 agonist-, forskolin-, and 8-bromo-cAMP-stimulated phosphorylation of DARPP-32. The ability of quinpirole to decrease D1-stimulated DARPP-32 phosphorylation was calcium-dependent and was blocked by the calcineurin inhibitor cyclosporin A, suggesting that the D2 effect involved an increase in intracellular calcium and activation of calcineurin. In support of this interpretation, Ca2+-free/EGTA medium induced a greater than 60-fold increase in DARPP-32 phosphorylation and abolished the ability of quinpirole to dephosphorylate DARPP-32. The antipsychotic drug raclopride, a selective D2 receptor antagonist, increased phosphorylation of DARPP-32 under basal conditions and in D2 agonist-treated slices. The results of this study demonstrate that dopamine exerts a bidirectional control on the state of phosphorylation of DARPP 32. PMID- 9334391 TI - Nervous system-specific expression of a novel serine protease: regulation in the adult rat spinal cord by excitotoxic injury. AB - A full-length cDNA clone of a previously unidentified serine protease, myelencephalon-specific protease (MSP), has been isolated by using a PCR cloning strategy and has been shown to be expressed in a nervous system and spinal cord specific pattern. Sequence analysis demonstrated that MSP is most similar in sequence to neuropsin, trypsin, and tissue kallikrein and is predicted to have trypsin-like substrate specificity. MSP mRNA was found to be approximately 10 fold greater in the CNS of the rat and human, as compared with most peripheral tissues, and within the CNS was found to be highest by a factor of four in the medulla oblongata and spinal cord. Levels of mRNA encoding tissue plasminogen activator (tPA) also were elevated in the spinal cord but were more widespread in peripheral tissues as compared with MSP. In the adult rat lumbosacral spinal cord, in situ localization of MSP mRNA demonstrated 2-fold higher levels in the white, as compared with the gray, matter. MSP mRNA expression was shown to increase 3-fold in the white matter and 1.5-fold in the gray laminae at 72 hr after intraperitoneal injection of the AMPA/kainate glutamate receptor-specific agonist, kainic acid (KA). MSP mRNA remained elevated in the ventral gray matter, including expression associated with the motor neurons of lamina IX, at 7 d after the initial excitotoxic insult. Together, these observations indicate that MSP is in a position to play a fundamental role in normal homeostasis and in the response of the spinal cord to injury. PMID- 9334392 TI - Inhibition of synaptic transmission by neuropeptide Y in rat hippocampal area CA1: modulation of presynaptic Ca2+ entry. AB - Neuropeptide Y (NPY) agonists inhibit glutamate release by a presynaptic action at the CA3-CA1 synapse of rat hippocampus. We have examined the relationship between [Capre]t via presynaptic, voltage-dependent calcium channels (VDCCs), measured optically by using the fluorescent calcium indicator fura-2, and transmitter release, measured electrophysiologically. Activation of presynaptic NPY Y2 receptors reduced [Capre]t and thereby inhibited synaptic transmission. Multiple calcium channels are involved in synaptic transmission at this synapse. Activation of Y2 receptors inhibits N-type, P/Q-type, and unidentified presynaptic VDCCs. The inhibition of each of these calcium channel types contributes to the reduction of [Capre]t by Y2 receptors. Activation of adenosine receptors fully occluded the inhibition of presynaptic calcium influx by Y2 receptors but not the inhibition by GABAB receptors, suggesting a convergent action for Y2 and adenosine receptors, probably by coupling to the same G protein. PMID- 9334393 TI - The actin-severing protein gelsolin modulates calcium channel and NMDA receptor activities and vulnerability to excitotoxicity in hippocampal neurons. AB - Calcium influx through NMDA receptors and voltage-dependent calcium channels (VDCC) mediates an array of physiological processes in neurons and may also contribute to neuronal degeneration and death in neurodegenerative conditions such as stroke and severe epileptic seizures. Gelsolin is a Ca2+-activated actin severing protein that is expressed in neurons, wherein it may mediate motility responses to Ca2+ influx. Primary hippocampal neurons cultured from mice lacking gelsolin exhibited decreased actin filament depolymerization and enhanced Ca2+ influx after exposure to glutamate. Whole-cell patch-clamp analyses showed that currents through NMDA receptors and VDCC were enhanced in hippocampal neurons lacking gelsolin, as a result of decreased current rundown; kainate-induced currents were similar in neurons containing and lacking gelsolin. Vulnerability of cultured hippocampal neurons to glutamate toxicity was greater in cells lacking gelsolin. Seizure-induced damage to hippocampal pyramidal neurons was exacerbated in adult gelsolin-deficient mice. These findings identify novel roles for gelsolin in controlling actin-mediated feedback regulation of Ca2+ influx and in neuronal injury responses. The data further suggest roles for gelsolin and the actin cytoskeleton in both physiological and pathophysiological events that involve activation of NMDA receptors and VDCC. PMID- 9334394 TI - Amyloid beta-protein (Abeta) 1-40 but not Abeta1-42 contributes to the experimental formation of Alzheimer disease amyloid fibrils in rat brain. AB - Two major C-terminal variants ending at Val40 and Ala42 constitute the majority of amyloid beta-protein (Abeta), which undergoes postsecretory aggregation and deposition in the Alzheimer disease (AD) brain. To probe the differential pathobiology of the two Abeta variants, we used an in vivo paradigm in which freshly solubilized Abeta1-40 or Abeta1-42 was injected into rat brains, followed by examination using Congo red birefringence, Abeta immunohistochemistry, and electron microscopy. In the rat brain, soluble Abeta 1-40 and Abeta1-42 formed aggregates, and the Abeta1-40 but not the Abeta1-42 aggregates showed Congo red birefringence. Electron microscopy revealed that the Abeta1-40 aggregates contained fibrillar structures similar to the amyloid fibrils of AD, whereas the Abeta1-42 aggregates contained nonfibrillar amorphous material. Preincubation of Abeta1-42 solution in vitro led to the formation of birefringent aggregates, and after injection of the preincubated Abeta1-42, the aggregates remained birefringent in the rat brain. Thus, a factor or factors might exist in the rat brain that inhibit the fibrillar assembly of soluble Abeta1-42. To analyze the postsecretory processing of Abeta, we used the same in vivo paradigm and showed that Abeta1-40 and Abeta1-42 were processed at their N termini to yield variants starting at pyroglutamate, and at their C termini to yield variants ending at Val40 and at Val39. Thus the normal rat brain could produce enzymes that mediate the conversion of Abeta 1-40/1-42 into processed variants similar to those in AD. This experimental paradigm may facilitate efforts to elucidate mechanisms of Abeta deposition evolving into amyloid plaques in AD. PMID- 9334395 TI - Acetylcholine receptors in innervated muscles of dystrophic mdx mice degrade as after denervation. AB - Acetylcholine receptors (AChRs) are present at the top of the postsynaptic membrane of the neuromuscular junction (NMJ) at very high density, possibly anchored to cytoskeletal elements. The present study investigated whether AChR degradation is affected in animals lacking dystrophin, a protein that is an integral part of the cytoskeletal complex and is missing in Duchenne muscular dystrophy. The animal model for Duchenne muscular dystrophy, the mutant mdx mouse, was used to determine whether disruption of the cytoskeleton, caused by the absence of dystrophin, affects AChR degradation. Of the two populations of junctional AChRs, Rs (expressed in innervated adult muscles) and Rr (expressed in embryonic or denervated muscles), only Rs are affected in mdx animals. In innervated mdx soleus, diaphragm, and sternomastoid muscles, the AChRs have an accelerated degradation rate (t1/2 of approximately 3-5 d), similar to that acquired by Rs in control muscles after denervation. The Rs in mdx NMJs do not accelerate further when the muscles are denervated. The absence of dystrophin does not affect the degradation rate of the Rr AChRs (t1/2 of 1 d), which are expressed after denervation in mdx as in control muscles. These results suggest that dystrophin or an intact cytoskeletal complex may be required for neuronal stabilization of Rs receptors at the adult neuromuscular junctions. PMID- 9334396 TI - Neuronal alpha-bungarotoxin receptors differ structurally from other nicotinic acetylcholine receptors. AB - We have characterized the alpha-bungarotoxin receptors (BgtRs) found on the cell surface of undifferentiated pheochromocytoma (PC12) cells. The PC12 cells express a homogeneous population of alpha7-containing receptors that bind alpha-Bgt with high affinity (Kd = 94 pM). The BgtRs mediate most of the response elicited by nicotine, because the BgtR-specific antagonists methyllycaconitine and alpha-Bgt block approximately 90% of the whole-cell current. The binding of nicotinic agonists to cell-surface BgtRs was highly cooperative with four different agonists showing Hill coefficients in the range of 2.3-2.4. A similar agonist binding cooperativity was observed for BgtR homomers formed from chimeric alpha7/5HT3 subunits expressed in tsA 201 cells. Two classes of agonist binding sites, in the ratio of 4:1 for PC12 cell BgtRs and 3:1 for alpha7/5HT3 BgtRs, were revealed by bromoacetylcholine alkylation of the reduced sites on both PC12 BgtRs and alpha7/5HT3 BgtRs. We conclude from this data that PC12 BgtRs and alpha7/5HT3 homomers contain at least three distinguishable agonist binding sites and thus are different from other nicotinic receptors. PMID- 9334397 TI - Alternative splicing in the pore-forming region of shaker potassium channels. AB - We have cloned cDNAs for the shaker potassium channel gene from the spiny lobster Panulirus interruptus. As previously found in Drosophila, there is alternative splicing at the 5' and 3' ends of the coding region. However, in Panulirus shaker, alternative splicing also occurs within the pore-forming region of the protein. Three different splice variants were found within the P region, two of which bestow unique electrophysiological characteristics to channel function. Pore I and pore II variants differ in voltage dependence for activation, kinetics of inactivation, current rectification, and drug resistance. The pore 0 variant lacks a P region exon and does not produce a functional channel. This is the first example of alternative splicing within the pore-forming region of a voltage dependent ion channel. We used a recently identified potassium channel blocker, kappa-conotoxin PVIIA, to study the physiological role of the two pore forms. The toxin selectively blocked one pore form, whereas the other form, heteromers between the two pore forms, and Panulirus shal were not blocked. When it was tested in the Panulirus stomatogastric ganglion, the toxin produced no effects on transient K+ currents or synaptic transmission between neurons. PMID- 9334398 TI - Kappa2 opioid receptors in limbic areas of the human brain are upregulated by cocaine in fatal overdose victims. AB - Cocaine is thought to be addictive because chronic use leads to molecular adaptations within the mesolimbic dopamine (DA) circuitry that affect motivated behavior and emotion. Although the reinforcing effects of cocaine are mediated primarily by blocking DA reuptake into the presynaptic nerve terminal, reciprocal signaling between DA and endogenous opioids has important implications for cocaine dependence. The present study used the opioid antagonist 6 beta-[125iodo] 3,14-dihydroxy-17-cyclopropylmethyl-4,5 alpha-epoxymorphinan ([125I]IOXY) after pretreatment with the site-directed acylating agents 2-(p-ethoxybenzyl)-1 diethylaminoethyl-5-isothiocyanatobenzimid iazole -HCl (mu-selective) and N phenyl-N-[1-(2-(4-isothiocyanato)-phenethyl)-4-piperidinyl]-p ropana mide-HCl (delta-selective) to examine the effect of cocaine exposure on the distribution and density of kappa2 receptors in autopsy studies of human cocaine fatalities. The selective labeling of the kappa2 receptor subtype was demonstrated by competition binding studies, which gave a pharmacological signature (IOXY >/= (+) bremazocine >> U50,488 >/= U69,593) distinct from either the kappa1 or kappa3 receptor subtypes. Visualization of [125I]IOXY labeling revealed that kappa2 receptors localize to mesocortical and subcortical limbic areas, including the cingulate, entorhinal, insular, and orbitofrontal cortices and the nucleus accumbens and amygdala. The number of kappa2 receptors in the nucleus accumbens and other limbic brain regions from cocaine fatalities was increased twofold as compared with age-matched and drug-free control subjects. Cocaine overdose victims, who experienced paranoia and marked agitation before death, also had elevated densities of kappa2 receptors in the amygdala. These findings demonstrate for the first time that kappa2 receptor numbers are upregulated by cocaine exposure. The molecular adaptation of kappa2 receptor numbers may play a role in the motivational incentive associated with episodes of binge cocaine use and in the dysphoria that follows abrupt cocaine withdrawal. PMID- 9334399 TI - Characterization of delayed rectifier Kv channels in oligodendrocytes and progenitor cells. AB - We examined the molecular identity of K+ channel genes underlying the delayed rectifier (IK) in differentiated cultured oligodendrocytes (OLGs) and oligodendrocyte progenitor (OP) cells. Using reverse transcription-PCR cloning, we found that OP cells and OLGs expressed multiple Kv transcripts, namely Kv1.2, Kv1.4, Kv.1.5, and Kv1.6. Immunocytochemical and Western blot analyses revealed that Kv1.5 and Kv1.6 as well as Kv1.2 and Kv1.4 channel proteins could be detected in these cells, but definitive evidence for functional K+ channel expression was obtained only for the Kv1.5 channel. In addition, mRNA and immunoreactive protein levels of both Kv1.5 and Kv1.6 channels were significantly lower in differentiated OLGs when compared with levels in OP cells. Proliferation of OP cells was inhibited by K+ channel blockers, but not by incubation with either Kv1.5 or Kv1.6 antisense oligonucleotides. We conclude that (1) IK in OP cells and OLGs is encoded partly by Kv1.5 subunits, possibly forming heteromultimeric channels with Kv1.6 or other Kv subunits; and (2) inhibition of Kv1.5 or Kv1.6 channel expression alone does not prevent mitogenesis. Concomitant inhibition of other Kv channels underlying IK may be necessary for OP cells to exit from cell cycle. PMID- 9334400 TI - Association and colocalization of the Kvbeta1 and Kvbeta2 beta-subunits with Kv1 alpha-subunits in mammalian brain K+ channel complexes. AB - The differential expression and association of cytoplasmic beta-subunits with pore-forming alpha-subunits may contribute significantly to the complexity and heterogeneity of voltage-gated K+ channels in excitable cells. Here we examined the association and colocalization of two mammalian beta-subunits, Kvbeta1 and Kvbeta2, with the K+ channel alpha-subunits Kv1.1, Kv1.2, Kv1.4, Kv1.6, and Kv2.1 in adult rat brain. Reciprocal coimmunoprecipitation experiments using subunit specific antibodies indicated that Kvbeta1 and Kvbeta2 associate with all the Kv1 alpha-subunits examined, and with each other, but not with Kv2.1. A much larger portion of the total brain pool of Kv1-containing channel complexes was found associated with Kvbeta2 than with Kvbeta1. Single- and multiple-label immunohistochemical staining indicated that Kvbeta1 codistributes extensively with Kv1.1 and Kv1.4 in cortical interneurons, in the hippocampal perforant path and mossy fiber pathways, and in the globus pallidus and substantia nigra. Kvbeta2 codistributes extensively with Kv1.1 and Kv1.2 in all brain regions examined and was strikingly colocalized with these alpha-subunits in the juxtaparanodal region of nodes of Ranvier as well as in the axons and terminals of cerebellar basket cells. Taken together, these data provide a direct demonstration that Kvbeta1 and Kvbeta2 associate and colocalize with Kv1 alpha subunits in native tissues and provide a biochemical and neuroanatomical basis for the differential contribution of Kv1 alpha- and beta-subunits to electrophysiologically diverse neuronal K+ currents. PMID- 9334401 TI - OSM-9, a novel protein with structural similarity to channels, is required for olfaction, mechanosensation, and olfactory adaptation in Caenorhabditis elegans. AB - Although cyclic nucleotide-gated channels mediate sensory transduction in olfaction and vision, other forms of sensory transduction are independent of these channels. Caenorhabditis elegans cyclic nucleotide-gated channel mutants respond normally to some olfactory stimuli and to osmotic stimuli, suggesting that these chemosensory responses use an alternative sensory transduction pathway. One gene that may act in this pathway is osm-9, which is required for each of these responses as well as a mechanosensory response to nose touch. osm-9 encodes a protein with ankyrin repeats and multiple predicted transmembrane domains that has limited similarity to the Drosophila phototransduction channels transient receptor potential (TRP) and TRP-like (TRPL). The sequence of OSM-9 and other TRP-like genes reveals a previously unsuspected diversity of mammalian and invertebrate genes in this family. osm-9 is required for the activity of the predicted G-protein-coupled odorant receptor ODR-10, which acts in the AWA olfactory neurons; its similarity to other G-protein-regulated transduction channels suggests that OSM-9 is involved in AWA signaling. osm-9:: GFP fusion genes are expressed in a subset of chemosensory, mechanosensory, and osmosensory neurons. osm-9 also affects olfactory adaptation within neurons that require the cyclic nucleotide-gated channel for olfaction; in these neurons, the gene has a regulatory function and not a primary role in sensory transduction. PMID- 9334402 TI - Analysis of rat vestibular hair cell development and regeneration using calretinin as an early marker. AB - Despite increased interest in inner ear hair cell regeneration, it is still unclear what exact mechanisms underlie hair cell regeneration in mammals because of our limited understanding of hair cell development and the lack of specific hair cell markers. In this report, we studied hair cell development using immunohistochemistry on sections prepared from embryonic day (E) 13 to postnatal day 7 rat inner ear tissues. Of many epithelial, neuronal, and glial markers, we found that calcium-binding protein antibodies recognizing calretinin, calmodulin, or parvalbumin labeled immature hair cells in rat vestibular end organs. In particular, calretinin antiserum labeled the initial differentiating hair cells at E15, a stage immediately after the terminal mitosis of hair cell progenitors. The selective immunoreactivity of postmitotic presumptive hair cells, but not supporting cells or peripheral epithelial cells, was confirmed in utricular epithelial sheet cultures. Double labeling with calretinin and bromodeoxyuridine antibodies in long-term cultures showed that only a few mitotic utricular supporting cells became calretinin positive. Thus, although proliferation mediated regeneration of new hair cells might directly contribute to hair cell regeneration in rat utricles after injury, it is very limited. In addition, double labeling with calretinin and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) revealed that differentiated hair cells underwent apoptosis during normal development at late embryonic and early postnatal stages in vivo and in vitro. Therefore, these experiments lay the groundwork for the time course of differentiation, regeneration, and apoptosis of mammalian vestibular hair cells. This work also suggests that calcium-binding proteins are useful markers for studies on inner ear hair cell differentiation and regeneration. PMID- 9334403 TI - Angiotensin II AT1A receptor mRNA expression is induced by estrogen-progesterone in dopaminergic neurons of the female rat arcuate nucleus. AB - Brain angiotensin II (Ang II) inhibits pituitary prolactin release by an indirect mechanism requiring stimulation of dopamine formation and release. We report that [125I]Sar1-Ang II binding to AT1 receptors and AT1A receptor mRNA expression increase selectively in the dorsomedial arcuate nucleus of 17beta-estradiol primed ovariectomized rats after treatment with progesterone. In hormone-treated rats, arcuate nucleus AT1A receptor mRNA expression is associated with tyrosine hydroxylase-positive neurons. No AT1A receptor mRNA was detected in tyrosine hydroxylase-positive cells of the arcuate nucleus of intact male rats. Conversely, in the anterior pituitary, where local or circulating Ang II stimulates prolactin release, [125I]Sar1-Ang II binding to AT1 receptors and AT1B receptor mRNA expression are decreased in 17beta-estradiol/progesterone-treated ovariectomized rats. Thus, AT1A receptors in the dorsal arcuate nucleus and AT1B receptors in the anterior pituitary are regulated inversely by estrogen/progesterone treatment, supporting the hypothesis of a dual role for brain and pituitary Ang II on prolactin release. The colocalization of AT1A receptor mRNA and tyrosine hydroxylase in neurons of the arcuate nucleus furthermore indicates that within this area central Ang II acts directly on dopaminergic neurons. These results support the hypothesis that central Ang II inhibits pituitary prolactin release indirectly via modulation of dopaminergic activity in the arcuate nucleus. PMID- 9334404 TI - Activation of ErbB2 during wallerian degeneration of sciatic nerve. AB - We used anti-phosphopeptide-immunodetecting antibodies as immunohistochemical reagents to define the location and activity state of p185(erbB2) during Wallerian degeneration. Nerve damage induces a phosphorylation event at Y1248, a site that couples p185(erbB2) to the Ras-Raf-MAP kinase signal transduction pathway. Phosphorylation of p185(erbB2) occurs within Schwann cells and coincides in time and space with Schwann cell mitotic activity, as measured by bromodeoxyuridine uptake. These visual images of receptor autophosphorylation link activation of p185(erbB2) to the Schwann cell proliferation that accompanies nerve regeneration. PMID- 9334405 TI - Expression of neural RNA-binding proteins in the postnatal CNS: implications of their roles in neuronal and glial cell development. AB - There is an increasing interest in the role of RNA-binding proteins during neural development. Mouse-Musashi-1 (m-Msi-1) is a mouse neural RNA-binding protein with sequence similarity to Drosophila musashi (d-msi), which is essential for neural development. m-Msi-1 is highly enriched in neural precursor cells that are capable of generating both neurons and glia during embryonic CNS development. The present study characterized m-Msi-1-expressing cells in the postnatal and adult CNS. Postnatally, m-Msi-1 was expressed in proliferative neuronal precursors in the external granule cell layer of the cerebellum and in the anterior corner of the subventricular zone of the lateral ventricles. In gliogenesis, the persistent expression of m-Msi-1 was observed in cells of the astrocyte lineage ranging from proliferative glial precursors in the subventricular zone (SVZ) to differentiated astrocytes in the parenchyma. In addition, we showed that m-Msi-1 was still expressed in proliferating cells in the adult SVZ, which may contain neural precursor or stem cells. Another neural RNA-binding protein Hu (the mammalian homolog of a Drosophila neuronal RNA-binding protein Elav) was present in postmitotic neurons throughout the development of the CNS, and its pattern of expression was compared with that of m-Msi-1. These observations imply that these two RNA-binding proteins may be involved in the development of neurons and glia by regulating gene expression at the post-transcriptional level. PMID- 9334406 TI - Origin and route of tangentially migrating neurons in the developing neocortical intermediate zone. AB - Neuroblasts produced in the ventricular zone of the neocortex migrate radially and form the cortical plate, settling in an inside-out order. It is also well known that the tangential cell migration is not negligible in the embryonic neocortex. To have a better understanding of the tangential cell migration in the cortex, we disturbed the migration by making a cut in the neocortex, and we labeled the migrating cells with 1,1'-dioctodecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI) in vivo and in vitro. We also determined the birth dates of the cells. Disturbance of tangential cell migration caused an accumulation and disappearance of microtubule-associated protein 2 immunoreactive (MAP2-IR) cells on the ventral and dorsal side of the cut, respectively, which indicated that most of the MAP2-IR cells in the intermediate zone (IZ) were migrating toward the dorsal cortex. The DiI injection study in vivo confirmed the tendency of the direction of cell migration and suggested the origin of the cells to be in the lateral ganglionic eminence (LGE). DiI injection into the LGE in vitro confirmed that the LGE cells cross the corticostriatal boundary and enter the IZ of the neocortex. The migrating cells acquired multipolar shape in the IZ of the dorsal cortex and seemed to reside there. A 5 bromo-deoxyuridine incorporation study revealed that the migrating MAP2-IR cells in the IZ were early-generated neurons. We concluded that the majority of tangentially migrating cells were generated in the LGE and identified as a distinct population that was assumed not to have joined the cortical plate. PMID- 9334408 TI - A role for collapsin-1 in olfactory and cranial sensory axon guidance. AB - Collapsin-1 is a member of the semaphorin family of signaling molecules that acts as a repellent for growing spinal sensory axons. We have constructed a chimeric collapsin-1/alkaline phosphatase probe to visualize putative collapsin-1 receptors in vitro and in situ. As predicted by the activity profile of collapsin 1, the probe binds spinal sensory tracts, ventral spinal roots, and the sympathetic chain but does not bind retinal axons. In addition, we find that the probe binds sensory axons arising from the olfactory epithelium and some, but not all, cranial sensory nerves. As predicted by these binding studies, in vitro assays demonstrate that primary olfactory sensory, trigeminal, and jugular ganglion growth cones collapse in the presence of soluble collapsin-1. Comparing the expression pattern of collapsin-1 with the trajectories of collapsin-1 responsive axons suggests that in both the spinal cord and the olfactory bulb, collapsin-1 prevents premature entry of sensory axons into their target and helps determine the final location of sensory terminations. PMID- 9334407 TI - Modulation of intrinsic circuits by serotonin 5-HT3 receptors in developing ferret visual cortex. AB - Serotonergic projections are widespread in the developing neocortex, but their functions are obscure. The effects of 5-HT3 receptor agonists on cortical circuit response properties were studied in slices of ferret primary visual cortex using high-speed optical imaging of voltage-sensitive dye signals and whole-cell patch clamp recording. Activation of the 5-HT3 receptor decreased the amplitude and lateral extent of excitation throughout postnatal development. This effect peaks after eye opening, which indicates a function for serotonergic modulation of circuit responses during the period of refinement of cortical connections. Whole cell patch-clamp recordings from single neurons revealed that synaptic responses evoked by white matter stimulation were reduced by 5-HT3 receptor agonists, whereas the frequency of spontaneous GABAergic synaptic currents was enhanced dramatically. This indicates that the modulation of spontaneous synaptic activity by fast-acting serotonin receptors is reflected in an inhibition of the circuit response, in line with the notion of background synaptic activity altering the spatiotemporal integration properties of cortical cells by changing their membrane potential and their electrotonic structure. These mechanisms may regulate the response properties of intrinsic circuits in both the adult and developing neocortex. PMID- 9334409 TI - Fast synaptic signaling by nicotinic acetylcholine and serotonin 5-HT3 receptors in developing visual cortex. AB - Cholinergic and serotonergic fiber systems invade the developing visual cortex several weeks before eye opening; both transmitters have been implicated in plasticity of neocortical circuits. These transmitters have been presumed to act predominantly through second messenger-coupled receptors, because fast cholinergic or serotonergic neurotransmission has never been observed in neocortex. However, acetylcholine and serotonin also act on ligand-gated ion channels; the nicotinic acetylcholine receptor and the serotonin 5-HT3 receptor, respectively. Here, using whole-cell patch-clamp techniques in developing ferret visual cortex, we pharmacologically isolated fast, spontaneous, and evoked cholinergic and serotonergic synaptic events in pyramidal cells and interneurons of all cortical layers. The number of cells receiving such inputs increased with the ingrowth of thalamic afferents, and the frequencies of the spontaneous events increased at eye opening. Thus, both acetylcholine and serotonin can mediate fast synaptic transmission in the visual cortex; the early onset of these mechanisms suggests a role during initial stages of circuit formation and during subsequent experience-dependent remodeling of cortical connections. PMID- 9334410 TI - Glutamate transporter protein subtypes are expressed differentially during rat CNS development. AB - Extracellular glutamate concentrations are regulated by glial and neuronal transporter proteins. Four glutamate transporter subtypes have been identified in rat brain; GLAST and GLT-1 are primarily astrocytic, whereas EAAC1 and EAAT4 are neuronal. Using immunoblotting and immunohistochemistry with subtype-specific antipeptide antibodies, we examined the protein expression and regional and cellular localization of each glutamate transporter subtype in embryonic and postnatal rat CNS. Each transporter had a specific pattern of expression. GLAST immunoreactivity was low prenatally but became enriched in cerebellar Bergmann glia early postnatally and then was also present in forebrain later postnatally. The post-translational modification of GLAST was unique among the subtypes; glycosylated GLAST increased with maturation, whereas nonglycosylated protein decreased in abundance postnatally. GLT-1 was present in fetal brain and spinal cord, with expression progressively increasing to adult levels throughout the neuraxis by postnatal day 26. Transient expression of GLT-1 immunoreactivity along axonal pathways was observed prenatally, in contrast to the exclusive localization of GLT-1 to astrocytes in the adult CNS. EAAC1, localized to neurons, was enriched in forebrain, diencephalon, and hindbrain during prenatal and postnatal development. EAAC1 expression was greater in newborn brain compared with adult brain. EAAT4 had a region-specific distribution; EAAT4 was mainly in cerebellum, localized to Purkinje cells, with much lower levels in forebrain. EAAT4 levels increased in cerebellum with age. We conclude that during CNS development the expression of glutamate transporter subtypes is differentially regulated, regionally segregated, and coordinated. PMID- 9334411 TI - Activity-dependent regulation of NMDAR1 immunoreactivity in the developing visual cortex. AB - NMDA receptors have been implicated in activity-dependent synaptic plasticity in the developing visual cortex. We examined the distribution of immunocytochemically detectable NMDAR1 in visual cortex of cats and ferrets from late embryonic ages to adulthood. Cortical neurons are initially highly immunostained. This level declines gradually over development, with the notable exception of cortical layers 2/3, where levels of NMDAR1 immunostaining remain high into adulthood. Within layer 4, the decline in NMDAR1 immunostaining to adult levels coincides with the completion of ocular dominance column formation and the end of the critical period for layer 4. To determine whether NMDAR1 immunoreactivity is regulated by retinal activity, animals were dark-reared or retinal activity was completely blocked in one eye with tetrodotoxin (TTX). Dark rearing does not cause detectable changes in NMDAR1 immunoreactivity. However, 2 weeks of monocular TTX administration decreases NMDAR1 immunoreactivity in layer 4 of the columns of the blocked eye. Thus, high levels of NMDAR1 immunostaining within the visual cortex are temporally correlated with ocular dominance column formation and developmental plasticity; the persistence of staining in layers 2/3 also correlates with the physiological plasticity present in these layers in the adult. In addition, visual experience is not required for the developmental changes in the laminar pattern of NMDAR1 levels, but the presence of high levels of NMDAR1 in layer 4 during the critical period does require retinal activity. These observations are consistent with a central role for NMDA receptors in promoting and ultimately limiting synaptic rearrangements in the developing neocortex. PMID- 9334413 TI - Developmental synaptic depression underlying reorganization of visceral reflex pathways in the spinal cord. AB - During development, neuronal connectivity has a remarkable plasticity. Synaptic refinement in the spinal autonomic nucleus might be involved in the elimination of primitive segmental reflexes and the emergence of mature spinobulbospinal reflexes, which occurs a few weeks after birth. To address this possibility, we examined the postnatal changes of segmental excitatory synaptic transmission by applying the whole-cell recording technique to parasympathetic preganglionic neurons in slice preparations of the rat lumbosacral spinal cord. The mean magnitude of unitary excitatory synaptic currents evoked in preganglionic neurons by stimulation of single interneurons remained unchanged during the first two postnatal weeks but was reduced by 50% during the third postnatal week. This reduction in synaptic efficacy was associated with a decrease in the amount of transmitter release from interneurons. Moreover, this developmental depression of segmental synaptic transmission was prevented by spinal cord transection at the thoracic level on postnatal day 14. Thus, developmental modification of excitatory synapses on preganglionic neurons appears to be attributable to competition between segmental interneuronal and descending bulbospinal inputs, which results in the developmental reorganization of parasympathetic excretory reflex pathways. PMID- 9334412 TI - Bradykinin-induced collapse of rat pheochromocytoma (PC12) cell growth cones: a role for tyrosine kinase activity. AB - Pathfinding of growing nerve processes is guided by extracellular guidance cues. Here we report growth cone collapse of NGF-differentiated PC12 cells in culture evoked by the neuropeptide bradykinin. The growth cone response is mediated by B2 bradykinin receptors. Two different effects were distinguished. (1) Disappearance of filopodia occurred together with a loss of fibrillar actin (F-actin) in the growth cones at picomolar concentrations of bradykinin. The relative F-actin content was measured by means of rhodamine-phalloidin fluorescence using confocal microscopy. (2) Bradykinin-induced Ca2+ release and retraction of the neurite occurred at nanomolar concentrations. Ca2+ responses at single growth cones were measured using a 1:1 mixture of fura-red and fluo-3 Ca2+-sensitive dyes. The [Ca2+]i rise is not a prerequisite for the observed effects, because F-actin loss and retraction occurred during inhibition of Ca2+ responses. In contrast, inhibition by genistein pointed to a tyrosine kinase activity in the bradykinin evoked cellular events. Subsequent analysis of phosphotyrosine proteins revealed that bradykinin stimulated tyrosine phosphorylation of the cytoskeleton associated protein paxillin and the nonreceptor protein tyrosine kinase pp60(c src). Paxillin and pp60(c-src) co-precipitated after bradykinin treatment. Immunostaining experiments showed punctate distribution of paxillin along PC12 neurites and in growth cones. Taken together, our data suggest that pp60(c-src) and paxillin are putative components of the intracellular signaling pathway of bradykinin-mediated neurite retraction and provide evidence for a crosstalk between G-protein- and tyrosine kinase-dependent pathways in these cellular events. PMID- 9334414 TI - Transition from growth cone to functional motor nerve terminal in Drosophila embryos. AB - As a motor axon grows from the CNS to its target muscle, the terminal has the form of a flattened growth cone with a planar central region, lamellipodia, and filopodia. A mature terminal usually has a stereotyped shape that may be elongated with varicosities, as in several invertebrate species, or have short branches with boutons, as in mammals. We examined in Drosophila the developmental changes between growth cone and mature terminal using ultrastructural and immunocytochemical methods. The transition period, which occurs 2-3 hr after the first growth cone reaches its target muscle, is marked by the formation of "prevaricosities," smoothly contoured enlargements of the axons at the point where the nerve trunk first contacts the muscle fiber (MF). There is a 15-30 min ventral-to-dorsal gradient in the formation of prevaricosities on the individual abdominal MFs. Multineuronal innervation of each MF has occurred by this time, and two or more different axons undergo prevaricosity formation while they are intimately intertwined at the nerve entry point (NEP). Presynaptic active zones, both nerve-nerve and nerve-muscle, occur within the prevaricosities along broad contact regions. Synaptotagmin immunoreactive clusters form concurrently. The first varicosities then develop as a result of constrictions of the larger prevaricosities rather than as enlargement of discrete portions of the filopodia or neurites. The prevaricosity stage therefore may include the key steps that lead to the differentiation of functional differences in terminal subtypes as well as those leading to the formation of a stable neuromuscular junction. PMID- 9334415 TI - Synaptic communication among hippocampal interneurons: properties of spontaneous IPSCs in morphologically identified cells. AB - The properties of spontaneous IPSCs (sIPSCs) recorded with whole-cell patch-clamp techniques were investigated in various anatomically identified hippocampal CA1 interneurons and were compared with those recorded in pyramidal cells. Neurons labeled with biocytin or neurobiotin were classified on the basis of their dendritic and axonal arborizations, leading to the identification of previously unknown interneuron types projecting to the dendritic region of pyramidal cells. In most interneurons, the average sIPSCs decayed slower than did those observed in pyramidal cells. The properties of sIPSCs were homogeneous within a given morphologically identified neuron type. Many interneurons had comparable somatic size, location, and dendritic arbor but displayed extremely different axonal projections paralleled by distinct sIPSC properties. Thus, physiological comparisons are only meaningful after the complete morphological identification of the recorded cells. The decay of sIPSCs matched for amplitudes and rise times could vary over 10-fold in a given interneuron, consistent with electrotonic filtering and possibly with different GABAA receptor subunit assemblies present at distinct synapses. Our findings demonstrate an extensive connectivity among hippocampal interneurons through GABAA synapses of various properties that may underlie complex network oscillations at different frequencies. PMID- 9334416 TI - cAMP response element-binding protein in the amygdala is required for long- but not short-term conditioned taste aversion memory. AB - In conditioned taste aversion (CTA) organisms learn to avoid a taste if the first encounter with that taste is followed by transient poisoning. The neural mechanisms that subserve this robust and long-lasting association of taste and malaise have not yet been elucidated, but several brain areas have been implicated in the process, including the amygdala. In this study we investigated the role of amygdala in general, and the cAMP response element-binding protein (CREB) in the amygdala in particular, in CTA learning and memory. Toward that end, we combined antisense technology in vivo with behavioral, molecular, and histochemical analysis. Local microinjection of phosphorothioate-modified oligodeoxynucleotides (ODNs) antisense to CREB into the rat amygdala several hours before CTA training transiently reduced the level of CREB protein during training and impaired CTA memory when tested 3-5 d later. In comparison, sense ODNs had no effect on memory. The effect of antisense was not attributable to differential tissue damage and was site-specific. CREB antisense in the amygdala had no effect on retrieval of CTA memory once it had been formed, and did not affect short-term CTA memory. We propose that the amygdala, specifically the central nucleus, is required for the establishment of long-term CTA memory in the behaving rat; that the process involves long-term changes, subserved by CRE regulated gene expression, in amygdala neurons; and that the amygdala may retain some CTA-relevant information over time rather than merely modulating the gustatory trace during acquisition of CTA. PMID- 9334417 TI - Reduced levels of norepinephrine transporters in the locus coeruleus in major depression. AB - The norepinephrine transporter (NET) is a membrane protein responsible for termination of the action of synaptic norepinephrine and is a site of action of many drugs used to treat major depression. The present study determined whether the binding of [3H]nisoxetine to the NET is altered in the locus coeruleus (LC) in major depression, using brain tissue collected postmortem from subjects diagnosed with major depression and from age-matched normal control subjects. Thirteen of the 15 major depressive subjects studied died by suicide. The distribution of [3H]nisoxetine binding along the rostro-caudal axis of the nucleus was uneven and was paralleled by a similar uneven distribution of neuromelanin-containing cells in both major depressives and psychiatrically normal control subjects. The binding of [3H]nisoxetine to NETs in the midcaudal portion of the LC from major depressive subjects was significantly lower than that from age-matched, normal control subjects. The binding of [3H]nisoxetine to NETs in other regions of the LC was similar in major depressives and control subjects. In contrast to reductions in binding to NETs, there were no significant differences in the number of noradrenergic cells at any particular level of the LC between major depressives and normal control subjects. The decreased binding of [3H]nisoxetine to NETs in the LC in major depression may reflect a compensatory downregulation of this transporter protein in response to an insufficient availability of its substrate (norepinephrine) at the synapse. PMID- 9334418 TI - Nerve growth factor- and neurotrophin-3-induced changes in nociceptive threshold and the release of substance P from the rat isolated spinal cord. AB - Acute superfusion of nerve growth factor (NGF; 1-100 ng/ml) through a naive rat spinal cord preparation did not alter basal or electrically evoked release of substance P-like immunoreactivity (SP-LI). In contrast, neurotrophin-3 (NT-3; 1 100 ng/ml), although not modifying SP-LI basal outflow, dose-dependently inhibited the electrically evoked, but not capsaicin (10 nM)-induced, release of the peptide. This NT-3 (10 ng/ml)-induced inhibition persisted even in the presence of 100 ng/ml NGF in the perfusion fluid and was still significant when the evoked release of SP-LI was enhanced by a prolonged in vivo treatment with NGF. Co-superfusion with naloxone (0.1 microM), but not CGP 36742 (100 microM), a GABAB antagonist, prevented NT-3 (10 ng/ml) inhibition of SP-LI release. Basal and electrically evoked release of SP-LI from the rat spinal cord in vitro was not modified 24 hr after single systemic injection of either NGF (1 mg/kg) or NT 3 (10 mg/kg). At these time intervals from administration, NGF had induced thermal and mechanical hyperalgesia in the rat hindpaw, and NT-3 had induced mechanical, but not thermal, hypoalgesia. NT-3 administered six times over a 2 week period (at 1 mg/kg) did not alter thermal threshold but significantly reduced electrically evoked release of SP-LI from the spinal cord. An identical treatment regimen with 1 mg/kg NGF induced a significant increase in evoked release of SP-LI. However, this was not associated with a significant hyperalgesia. Although finding that NGF-induced hyperalgesia does not clearly correlate with changes in the release of SP-LI in the spinal cord, this study shows that NT-3 is an inhibitor of SP-LI release and suggests that this mechanism may be responsible for NT-3-induced antinociception. PMID- 9334419 TI - Circadian phase shifts to neuropeptide Y In vitro: cellular communication and signal transduction. AB - Mammalian circadian rhythms originate in the hypothalamic suprachiasmatic nuclei (SCN), from which rhythmic neural activity can be recorded in vitro. Application of neurochemicals can reset this rhythm. Here we determine cellular correlates of the phase-shifting properties of neuropeptide Y (NPY) on the hamster circadian clock in vitro. Drug or control treatments were applied to hypothalamic slices containing the SCN on the first day in vitro. The firing rates of individual cells were sampled on the second day in vitro. Control slices exhibited a peak in firing rate in the middle of the day. Microdrop application of NPY to the SCN phase advanced the time of peak firing rate. This phase-shifting effect of NPY was not altered by block of sodium channels with tetrodotoxin or block of calcium channels with cadmium and nickel, consistent with a direct postsynaptic site of action. Pretreatment with the glutamate receptor antagonists (DL-2-amino-5 phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione disodium) also did not alter phase shifts to NPY. Blocking GABAA receptors with bicuculline (Bic) had effects only at very high (millimolar) doses of Bic, whereas blocking GABAB receptors did not alter effects of NPY. Phase shifts to NPY were blocked by pretreatment with inhibitors of protein kinase C (PKC), suggesting that PKC activation may be necessary for these effects. Bathing the slice in low Ca2+/high Mg2+ can block phase shifts to NPY, possibly via a depolarizing action. A depolarizing high K+ bath can also block NPY phase shifts. The results are consistent with direct action of NPY on pacemaker neurons, mediated through a signal transduction pathway that depends on activation of PKC. PMID- 9334420 TI - Nerve growth factor treatment increases brain-derived neurotrophic factor selectively in TrkA-expressing dorsal root ganglion cells and in their central terminations within the spinal cord. AB - Using immunocytochemistry and in situ hybridization, we have examined the expression of brain-derived neurotrophic factor (BDNF) and of neurotrophin receptors in dorsal root ganglion cells. In the adult rat, BDNF mRNA and protein were found mainly in the subpopulation of cells that express the nerve growth factor (NGF) receptor trkA and the neuropeptide calcitonin gene-related peptide (CGRP). NGF increased BDNF within the trkA/CGRP cells to the extent that almost 90% of trkA cells contained BDNF mRNA after intrathecal NGF treatment, and 80-90% of BDNF-expressing cells contained trkA. Non-trkA cells that expressed BDNF included some trkC cells and some small cells that labeled with the lectin Griffonia simplicifolia IB4, a marker for cells that do not express trks. However, very few trkB cells expressed either BDNF mRNA or protein, and NGF did not increase BDNF expression in non-trkA cells. BDNF protein was anterogradely transported both peripherally and centrally. The central transport resulted in BDNF immunoreactivity in CGRP containing terminal arbors in the dorsal horn of the spinal cord, and this immunoreactivity was increased by NGF treatment. Electron microscopic analysis revealed that the BDNF immunoreactivity was present in finely myelinated and unmyelinated axons and in axon terminals, where it was most concentrated over dense-cored vesicles. Our data do not support an autocrine or paracrine role for BDNF within normal dorsal root ganglia, but indicate that BDNF may act as an anterograde trophic messenger. NGF levels in the periphery could influence dorsal horn neurons via release of BDNF from primary afferents. PMID- 9334421 TI - Persistent structural modifications in nucleus accumbens and prefrontal cortex neurons produced by previous experience with amphetamine. AB - Experience-dependent changes in behavior are thought to involve structural modifications in the nervous system, especially alterations in patterns of synaptic connectivity. Repeated experience with drugs of abuse can result in very long-lasting changes in behavior, including a persistent hypersensitivity (sensitization) to their psychomotor activating and rewarding effects. It was hypothesized, therefore, that repeated treatment with the psychomotor stimulant drug amphetamine, which produces robust sensitization, would produce structural adaptations in brain regions that mediate its psychomotor activating and rewarding effects. Consistent with this hypothesis, it was found that amphetamine treatment altered the morphology of neurons in the nucleus accumbens and prefrontal cortex. Exposure to amphetamine produced a long-lasting (>1 month) increase in the length of dendrites, in the density of dendritic spines, and in the number of branched spines on the major output cells of the nucleus accumbens, the medium spiny neurons, as indicated by analysis of Golgi-stained material. Amphetamine treatment produced similar effects on the apical (but not basilar) dendrites of layer III pyramidal neurons in the prefrontal cortex. The ability of amphetamine to alter patterns of synaptic connectivity in these structures may contribute to some of the long-term behavioral consequences of repeated amphetamine use, including amphetamine psychosis and addiction. PMID- 9334422 TI - Substantia nigra D1 receptors and stimulation of striatal cholinergic interneurons by dopamine: a proposed circuit mechanism. AB - Dopamine release can regulate striatal acetylcholine efflux in vivo through at least two receptor mechanisms: (1) direct inhibition by dopamine D2 receptors on the cholinergic neurons, and (2) excitation initiated by dopamine D1 receptors. The neuroanatomical locus of the latter population of D1 receptors and the pathway(s) involved in the expression of their influence are controversial issues. We have tested the hypothesis that D1 receptors in substantia nigra pars reticulata are involved in the excitatory component of dopaminergic actions on striatal acetylcholine output. In vivo microdialysis was used in awake rats. Infusion of the selective D1 receptor agonist R(+)-1-Phenyl-2,3,4,5-tetrahydro-1H 3-benzazepine-7,8-diol (SKF 38393) hydrochloride into pars reticulata of substantia nigra elicited a significant increase in striatal acetylcholine efflux. Likewise, D-amphetamine applied into pars reticulata of substantia nigra by reverse dialysis produced an elevation in acetylcholine output measured at a second microdialysis probe in the striatum. Application of D-amphetamine in the striatum by reverse dialysis elicited a decrease in striatal acetylcholine efflux that could be reversed subsequently by local application of D-amphetamine in substantia nigra pars reticulata. A 2 mg/kg intraperitoneal dose of D amphetamine, which has no net effect on striatal acetylcholine output under control conditions, elicited a significant decrease in acetylcholine efflux when the D1 receptor antagonist R(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4, 5 tetrahydro-1H-3-benzazepine (SCH 23390) hydrochloride was applied simultaneously via a second microdialysis probe in substantia nigra pars reticulata. Thus, an excitatory D1-mediated influence on striatal acetylcholine output is initiated in substantia nigra pars reticulata, and this influence contributes to the effects of indirect dopaminergic agonists such as D-amphetamine on striatal acetylcholine efflux. These results indicate an important role of somatodendritic dopamine release, in addition to nerve terminal dopamine release, in the regulation of activity in basal ganglia circuits. PMID- 9334423 TI - Neurturin and glial cell line-derived neurotrophic factor receptor-beta (GDNFR beta), novel proteins related to GDNF and GDNFR-alpha with specific cellular patterns of expression suggesting roles in the developing and adult nervous system and in peripheral organs. AB - Cloning strategies were used to identify a gene termed glial cell line-derived neurotrophic factor receptor-beta (GDNFR-beta) related to GDNFR-alpha. In situ hybridization was then used to map cellular expression of the GDNF-related trophic factor neurturin (NTN) and GDNFR-beta mRNA in developing and adult mice, and comparisons with GDNFR-alpha and RET were made. Neurturin is expressed in postnatal cerebral cortex, striatum, several brainstem areas, and the pineal gland. GDNFR-beta mRNA was more widely expressed in the developing and adult CNS, including cerebral cortex, cerebellum, thalamus, zona incerta, hypothalamus, brainstem, and spinal cord, and in subpopulations of sensory neurons and developing peripheral nerves. NTN colocalized with RET and GDNFR-alpha in ureteric buds of the developing kidney. The circular muscle layer of the developing intestines, smooth muscle of the urether, and developing bronchiolae also expressed NTN. GDNFR-beta was found in myenteric but not submucosal intestinal plexuses. In developing salivary glands NTN had an epithelial expression, whereas GDNFR-beta was expressed in surrounding tissue. Neurturin and GDNFR-beta were present in developing sensory organs. In the gonads, NTN appeared to be expressed in Sertoli cells and in the epithelium of the oviduct, whereas GDNFR-beta was expressed by the germ cell line. Our findings suggest multiple roles for NTN and GDNFR-beta in the developing and adult organism. Although NTN and GDNFR-beta expression patterns are sometimes complementary, this is not always the case, suggesting multiple modi operandi of GDNF and NTN in relation to RET and the two binding proteins, GDNFR-alpha and GDNFR-beta. PMID- 9334424 TI - Opposite modulation of opiate withdrawal behaviors on microinfusion of a protein kinase A inhibitor versus activator into the locus coeruleus or periaqueductal gray. AB - Chronic opiate administration upregulates the cAMP pathway in the locus coeruleus (LC). This adaptation is thought to increase the electrical excitability of LC neurons and contribute to the dramatic increase in LC firing induced by opioid receptor antagonists in opiate-dependent animals. The goal of the present study was to evaluate directly a role of the cAMP pathway in opiate withdrawal behaviors by studying, in vivo, whether withdrawal is influenced by intra-LC infusion of compounds known to activate or inhibit protein kinase A (PKA). Infusions into amygdala or periaqueductal gray (PAG) were studied for comparison. In one series of experiments the effect of intra-LC, intra-amygdala, or intra-PAG infusions of the PKA inhibitor Rp-cAMPS on naloxone-precipitated withdrawal from morphine was examined. Intra-LC infusions of Rp-cAMPS significantly attenuated several prominent behavioral signs of morphine withdrawal. Intra-PAG infusions of Rp-cAMPS also significantly attenuated opiate withdrawal behaviors, although different behaviors were affected. In contrast, intra-amygdala infusions of Rp cAMPS were without significant effect. In a second series of experiments the effect of intra-LC or intra-PAG infusions of the PKA activator Sp-cAMPS on behavior in nondependent drug-naive animals was determined. Sp-cAMPS infusions into either brain region induced a quasi-withdrawal syndrome, but the observed behaviors differed between the two groups. Analysis of the phosphorylation state of tyrosine hydroxylase, a well characterized substrate for PKA, confirmed the ability of Rp-cAMPS and Sp-cAMPS to inhibit and activate, respectively, PKA activity in vivo. Together, these data provide direct evidence for involvement of the cAMP-PKA system in the LC, as well as in the PAG, in opiate withdrawal and withdrawal-related behaviors. PMID- 9334425 TI - Supranormal stimulation of D1 dopamine receptors in the rodent prefrontal cortex impairs spatial working memory performance. AB - Although previous research has emphasized the beneficial effects of dopamine (DA) on functions of the prefrontal cortex (PFC), recent studies of animals exposed to mild stress indicate that excessive DA receptor stimulation may be detrimental to the spatial working memory functions of the PFC (Arnsten and Goldman-Rakic, 1990; Murphy et al., 1994, 1996a,b, 1997). In particular, these studies have suggested that supranormal stimulation of D1 receptors may contribute to the detrimental actions of DA in the PFC (Murphy et al., 1994, 1996a). The current study directly tested this hypothesis by examining the effects of infusing a full D1 receptor agonist, SKF 81297, into the PFC of rats performing a spatial working memory task, delayed alternation. SKF 81297 produced a dose-related impairment in delayed-alternation performance. The impairment was reversed by pretreatment with a D1 receptor antagonist, SCH 23390, consistent with drug actions at D1 receptors. SCH 23390 by itself had no effect on performance, although slightly higher doses impaired performance (Murphy et al., 1994, 1996a). There was a significant relationship between infusion location and drug efficacy; animals with cannulae anterior to the PFC were not impaired by SKF 81297 infusions. Taken together, these results demonstrate that supranormal D1 receptor stimulation in the PFC is sufficient to impair PFC working memory function. These cognitive data are consistent with recent electrophysiological studies of D1 receptor mechanisms affecting the PFC (Williams and Goldman-Rakic, 1995; Yang and Seamans, 1996). Increased D1 receptor stimulation during stress may serve to take the PFC "off line" to allow posterior cortical and subcortical structures to regulate behavior, but may contribute to the vulnerability of the PFC in many neuropsychiatric disorders. PMID- 9334426 TI - Rhinal cortex removal produces amnesia for preoperatively learned discrimination problems but fails to disrupt postoperative acquisition and retention in rhesus monkeys. AB - To test whether the rhinal cortex (i.e., entorhinal and perirhinal cortex) plays a time-limited role in information storage, eight rhesus monkeys were trained to criterion on two sets of 60 object discrimination problems, one set at each of two different time periods separated by 15 weeks. After the monkeys had learned both sets, two groups balanced for preoperative acquisition rates were formed. One group received bilateral ablation of the rhinal cortex (n = 4), and the other was retained as an unoperated control group (n = 4). After a 2 week rest period, monkeys were assessed for retention of the object discrimination problems. Retention was significantly poorer in monkeys with removals of the rhinal cortex relative to the controls (68 vs 91%). Although both groups showed slightly better retention of problems from the more recently learned set, there was no evidence of a differential effect of the cortical removal across sets (i.e., no temporal gradient). In addition, the monkeys with rhinal cortex lesions subsequently learned three new sets of 10 object discrimination problems as quickly as the controls did, thus ruling out the possibility of a gross impairment in visual perception or discrimination abilities. Furthermore, they retained these postoperatively learned object discriminations as well as the controls did. The findings indicate that the rhinal cortex is critical for the storage and/or retrieval of object discrimination problems that were learned up to 16 weeks before rhinal cortex ablation; however, in the absence of the rhinal cortex, efficient learning and retention of new discrimination problems can still occur. PMID- 9334427 TI - Response properties of corticotectal and corticostriatal neurons in the posterior lateral suprasylvian cortex of the cat. AB - Lateral suprasylvian cortex (LS) is an important source of visual projections to both the striatum and superior colliculus. Although these two LS efferent systems are likely to be involved in different aspects of visual processing, little is known about their functional properties. In the present experiments, 86 neurons in halothane-anesthetized, paralyzed cats were recorded along the posterior aspects of the medial and lateral banks of LS (PMLS and PLLS). Neurons were selected for analysis on the basis of antidromic activation from electrodes chronically implanted in the superior colliculus and caudate nucleus. The segregated nature of corticostriatal and corticotectal neurons was apparent; in no instance could a neuron be antidromically activated from both the superior colliculus and the caudate nucleus. Many common features were revealed between corticotectal and corticostriatal neurons; the majority of neurons in both populations were binocular and contralaterally dominant, showed similar responses to stationary flashed light, and expressed within-field spatial summation and surround inhibition. However, a number of information-processing features distinguished between corticotectal and corticostriatal neurons; the former were generally tuned to lower velocities than were the latter, and, for a given eccentricity in visual space, corticotectal neurons had smaller receptive fields than did corticostriatal neurons. Moreover, most corticotectal neurons displayed a marked preference for movements toward temporal visual space, whereas corticostriatal neurons revealed no specialization for a particular direction of movement. In addition, whereas corticotectal neurons were selective for receding stimuli, corticostriatal neurons were selective for approaching stimuli. The presence of these two corticofugal pathways is discussed in relation to their presumptive functional roles in the facilitation of attentive and orientation behaviors. PMID- 9334428 TI - Neuronal activity in monkey superior colliculus related to the initiation of saccadic eye movements. AB - The introduction of a temporal gap between the disappearance of an initially fixated target and the appearance of an eccentric saccadic target results in a general reduction of saccadic reaction times (SRTs)-the gap effect-and often in the production of express saccades, the latencies of which approach the conduction time of the shortest neural pathways from the retina to the eye muscles. We investigated saccade initiation by recording neuronal activity in the superior colliculus in monkeys performing the gap paradigm. Fixation-related neurons reduced their discharge rate during the gap period, regardless of the SRT. This reduction in activity is consistent with the hypothesized release of ocular fixation that facilitates premotor processes and may contribute to the gap effect. In addition to saccade-related discharges, many saccade-related neurons displayed phasic target-related responses and/or low-frequency preparatory activity during the gap period. The level of this preparatory activity correlated with both SRT and express saccade occurrence when the saccade was made into the response field of the neuron. Evidence indicates that advanced motor preparation is required for express saccade generation, which may be subserved by specific increases in the preparatory activity of saccade-related neurons. Increased preparatory activity may allow the target-related responses to trigger short latency express saccades directly. This study provides insights into the functional mechanism of saccade initiation and may be relevant to the generation of all voluntary motor responses. PMID- 9334429 TI - Role of dopamine D1 and D2 receptors in the nucleus accumbens in mediating reward. AB - The objectives of this study were to examine the involvement of D1 and D2 receptors within the nucleus accumbens (ACB) in mediating reinforcement. The intracranial self-administration (ICSA) of D1 and D2 agonists was used to determine whether activating D1 and/or D2 receptors within the ACB of Wistar rats is reinforcing. At concentrations of 0.25, 0.50, and 1.0 mM (25, 50, and 100 pmol/100 nl of infusion), neither the D1 agonist R(+)-1-phenyl-2,3,4,5-tetrahydro 1H-3-benzazepine-7,8-diol [SKF 38393 (SKF)] hydrochloride nor the D2 agonist (-) quinpirole (Quin) hydrochloride was self-administered into the shell region of the ACB. On the other hand, equimolar mixtures of SKF and Quin (SKF+Quin), at concentrations of 0.25, 0.50, and 1.0 mM each, were significantly self-infused into the ACB shell. The core region of the ACB did not support the ICSA of SKF+Quin at any of these concentrations. Rats increased lever pressing when the response requirement was increased from a fixed ratio 1 (FR1) to FR3, and they responded significantly more on the infusion lever than they did on the control lever. Coadministration of either 0.50 mM R(+)-7-chloro-8-hydroxy-3-methyl-1 phenyl-2,3,4, 5-tetrahydro-1H-3-benzazepine (SCH 23390) hydrochloride, a D1 antagonist, or 0.50 mM S(-)-sulpiride, a D2 antagonist, completely abolished the ICSA of the mixture of SKF+Quin (each at 0.50 mM) into the ACB shell. The present results suggest that concurrent activation of D1- and D2-type receptors in the shell of the ACB had a cooperative effect on DA-mediated reward processes. PMID- 9334430 TI - Status epilepticus-induced alterations in metabotropic glutamate receptor expression in young and adult rats. AB - In adult rats, kainic acid induces status epilepticus and delayed, selective cell loss of pyramidal neurons in the hippocampal CA3. In pup rats, kainate induces status epilepticus but not the accompanying neuronal cell death. The precise mechanisms underlying this age-dependent vulnerability to seizure-induced cell death are not understood. Metabotropic glutamate receptors (mGluRs) are developmentally and spatially regulated throughout the hippocampus and are implicated in seizure-induced damage. In the present study we used in situ hybridization to examine possible changes in mGluR expression at the level of the hippocampus after status epilepticus in postnatal day 10 (P10) pup and adult (P40) rats. Status epilepticus did not alter expression of mGluR1, mGluR3, or mGluR5 mRNAs. In pup and adult rats, status epilepticus induced a reduction in expression of mGluR2 mRNA in granule cells of the dentate gyrus. This change could lead to augmented glutamate release at mossy fiber synapses on CA3 pyramidal cells and thereby promote hyperexcitation. In pup but not adult rats, mGluR4 mRNA expression was enhanced in CA3 pyramidal neurons. Upregulation of presynaptic mGluR4 in pup CA3 neurons could lead to reduced transmitter release from CA3 axons, including recurrent collaterals, thereby reducing vulnerability of neonatal CA3 neurons to seizure-induced damage. These findings indicate that status epilepticus affects mGluR expression in a gene- and cell-specific manner, and that these changes vary with the developmental stage. PMID- 9334431 TI - Systemic morphine-induced Fos protein in the rat striatum and nucleus accumbens is regulated by mu opioid receptors in the substantia nigra and ventral tegmental area. AB - To characterize how systemic morphine induces Fos protein in dorsomedial striatum and nucleus accumbens (NAc), we examined the role of receptors in striatum, substantia nigra (SN), and ventral tegmental area (VTA). Morphine injected into medial SN or into VTA of awake rats induced Fos in neurons in ipsilateral dorsomedial striatum and NAc. Morphine injected into lateral SN induced Fos in dorsolateral striatum and globus pallidus. The morphine infusions produced contralateral turning that was most prominent after lateral SN injections. Intranigral injections of [D-Ala2, N-Me-Phe4, Gly-ol5]-enkephalin (DAMGO), a mu opioid receptor agonist, and of bicuculline, a GABAA receptor antagonist, induced Fos in ipsilateral striatum. Fos induction in dorsomedial striatum produced by systemic administration of morphine was blocked by (1) SN and VTA injections of the mu1 opioid antagonist naloxonazine and (2) striatal injections of either MK 801, an NMDA glutamate receptor antagonist, or SCH 23390, a D1 dopamine receptor antagonist. Fos induction in dorsomedial striatum and NAc after systemic administration of morphine seems to be mediated by dopamine neurons in medial SN and VTA that project to medial striatum and NAc, respectively. Systemic morphine is proposed to act on mu opioid receptors located on GABAergic interneurons in medial SN and VTA. Inhibition of these GABA interneurons disinhibits medial SN and VTA dopamine neurons, producing dopamine release in medial striatum and NAc. This activates D1 dopamine receptors and coupled with the coactivation of NMDA receptors possibly from cortical glutamate input induces Fos in striatal and NAc neurons. The modulation of target gene expression by Fos could influence addictive behavioral responses to opiates. PMID- 9334432 TI - Decreased frequency but not amplitude of quantal synaptic responses associated with expression of corticostriatal long-term depression. AB - We have investigated the site of expression of striatal long-term synaptic depression (LTD) using analysis of Sr2+-induced asynchronous release of quanta from stimulated synapses. The cumulative amplitude distribution of Sr2+-induced asynchronous synaptic responses overlaps with that of miniature EPSCs (mEPSCs), suggesting that Sr2+-induced asynchronous responses are quantal. Quantal amplitude at stimulated synapses is not significantly altered after LTD induction, whereas quantal frequency decreases after LTD induction. The decrease in quantal frequency is prevented when LTD expression is blocked by dialyzing 10 mM EGTA into the postsynaptic neuron. Our findings are most consistent with the idea that expression of striatal LTD involves decreased neurotransmitter release with no change in quantal amplitude, despite the fact that induction of striatal LTD involves postsynaptic mechanisms. PMID- 9334433 TI - Linearity and normalization in simple cells of the macaque primary visual cortex. AB - Simple cells in the primary visual cortex often appear to compute a weighted sum of the light intensity distribution of the visual stimuli that fall on their receptive fields. A linear model of these cells has the advantage of simplicity and captures a number of basic aspects of cell function. It, however, fails to account for important response nonlinearities, such as the decrease in response gain and latency observed at high contrasts and the effects of masking by stimuli that fail to elicit responses when presented alone. To account for these nonlinearities we have proposed a normalization model, which extends the linear model to include mutual shunting inhibition among a large number of cortical cells. Shunting inhibition is divisive, and its effect in the model is to normalize the linear responses by a measure of stimulus energy. To test this model we performed extracellular recordings of simple cells in the primary visual cortex of anesthetized macaques. We presented large stimulus sets consisting of (1) drifting gratings of various orientations and spatiotemporal frequencies; (2) plaids composed of two drifting gratings; and (3) gratings masked by full-screen spatiotemporal white noise. We derived expressions for the model predictions and fitted them to the physiological data. Our results support the normalization model, which accounts for both the linear and the nonlinear properties of the cells. An alternative model, in which the linear responses are subject to a compressive nonlinearity, did not perform nearly as well. PMID- 9334435 TI - Correction to "Carboplatin and Short-Infusion Paclitaxel in High Risk and Advanced-Stage Ovarian Carcinoma" PMID- 9334434 TI - Medial geniculate lesions block amygdalar and cingulothalamic learning-related neuronal activity. AB - This study assessed the role of the thalamic medial geniculate (MG) nucleus in discriminative avoidance learning, wherein rabbits acquire a locomotory response to a tone [conditioned stimulus (CS)+] to avoid a foot shock, and they learn to ignore a different tone (CS-) not predictive of foot shock. Limbic (anterior and medial dorsal) thalamic, cingulate cortical, or amygdalar lesions severely impair acquisition, and neurons in these areas develop training-induced activity (TIA): more firing to the CS+ than to the CS-. MG neurons exhibit TIA during learning and project to the amygdala. The MG neurons may supply afferents essential for amygdalar and cingulothalamic TIA and for avoidance learning. To test this hypothesis, bilateral electrolytic or excitotoxic ibotenic acid MG nuclear lesions were induced, and multiunit recording electrodes were chronically implanted into the anterior and posterior cingulate cortex, the anterior-ventral and medial-dorsal thalamic nuclei, and the basolateral nucleus of the amygdala before training. Learning was severely impaired and TIA was abolished in all areas in rabbits with lesions. Thus learning and TIA require the integrity of the MG nucleus. Only damage in the medial MG division was significantly correlated with the learning deficit. The lesions abolished the sensory response of amygdalar neurons, and they attenuated (but did not eliminate) the sensory response of cingulothalamic neurons, suggesting the existence of extra geniculate sources of auditory transmission to the cingulothalamic areas. PMID- 9334436 TI - [Dimorphic fungi: biochemical approach to their dimorphism]. AB - Dimorphism in pathogenic fungi is reviewed. The phenomenon is divided into four interwoven events: (a) perception of external stimuli by cellular sensors; (b) translation into a biochemical message; (c) alteration of the genomic expression, and (d) structural reorganization towards the morphological change. Experimental evidence is provided. Finally, the possibility that fungal dimorphism may have arisen multiple times throughout evolution, is discussed. PMID- 9334437 TI - [Purification and assay of chicken pepsin]. AB - The proteolitic enzyme pepsin (EC 3.4.23.1) was purified from chicken stomach by a modification of the method of Bohak (1970): after homogenazing the raw material, the zymogen was extracted with NaCl and NaHCO3, activated with 3N HCl and precipitated with NaCl (28% final concentration). The precipitate was lyophilised; fractions of it were suspended in 0.02N HCl. The solution was filtered through a column (2.6 x 80 cm) of Sephadex G-100 at an elution rate of 8 10 ml x cm-2 x h-1. Two protein peaks were obtained, the first one corresponding to the pepsin (2.0 mg/ml in pooled fractions). The milk clotting activity of the enzyme was determined on skimmed milk as a substrate (Berridge, 1955). Its proteolitic activity on the artificial substrate N-acetyl-L-phenylalanyl-L-3, 5 diiodo tyrosin (APD) also was determined (Rick-Fritsch, 1974). Mean clotting activity value was 5.52 UC, higher (P < 0.01) than that of the reference chymosin (0.64 UC). The activity with APD was unsatisfactory, due to very high absorbance values of the blanks. It is concluded, that the purification steps followed in this trial are simple and rapid, conferring a strong stimulus to using chicken pepsin as a clotting agent for the industrial production of pasteurized white cheese. PMID- 9334438 TI - [Pig lens cell membranes: ATPase activities stimulated by Na+]. PMID- 9334439 TI - [International Conference on Antibiotic Resistance]. PMID- 9334440 TI - [Ecology and postmodernity. An ethical point of view]. AB - This essay deals with critique of the "Modern Reason". Also, it aims to promote the debate about ethical and political issues of our technological/eco depredating and environmentally unsound model of development. We argue about ecology like a new knowledge point of view, from which to define criteria for the reformulation of the environment/society relationship, in a postmodern context. PMID- 9334441 TI - [The application of interactive multimedia software in taxonomy and biological diversity studies]. AB - To study biodiversity and to monitor changes in our biological environment high quality taxonomic and distributional data are imperative. At present access to species information and identification keys is limited by the fact that literature is scattered over a vast amount of sources. Exchange of biodiversity data between various researchers is hampered by the lack of universal documentation tools. The introduction of PC's in biological sciences rendered thousands of small and medium sized databases in hundreds of different incompatible formats. Modern multimedia computer techniques, allowing nearly unlimited storage of graphic information in addition to text, will facilitate distribution of easy accessible data. The Linnaeus II software package for biodiversity documentation is an answer to these needs as it presents a universal tool for biologists to document biodiversity and exchange standardized data. Regularly updated electronic monographs on CD-ROM will form the basis for modern, fast accessible libraries. PMID- 9334443 TI - Subcellular distribution and pharmacological characterization of muscarinic receptors in tracheal smooth muscle. AB - Subcellular fractions isolated from tracheal smooth muscle have been identified using biochemical markers and measuring the [3H]QNB muscarinic receptor binding activity in these fractions. This muscarinic receptor (mAchR) activity was slightly enriched 1.6 times in the crude mitochondrial fraction (M), 2.6 times in the crude microsomal fraction (P), and greatly enriched in the highly purified plasma membranes fractions, being 5.3 times in a heavy plasma membrane fraction designed as P2 and 9.1 times in a light plasma membrane fraction named P1 fraction. The muscarinic receptor subtypes present in the subcellular fractions were identified using competition experiments. The binding of five selective antagonists, pirenzepine, AF-DX 116, hexahydrodifenidol, methoctramine and 4-DAMP were examined. In this sense, the M1 antagonist pirenzepine showed pKi's values between 6.44-7.45 and the M2 antagonist AF-DX 116 showed pKi's values ranging from 6.75 to 7.45 being the lowest pKi's values here described. The antagonist hexahydrodifenidol showed higher affinities than pirenzepine-derivated compounds with pKi's values from 7.25 to 7.65. The antagonist 4-DAMP exhibited pKi's values from 8.18-8.41. Finally, methoctramine showed similar affinities as 4-DAMP, with pKi's ranging from 8.09 to 8.22 suggesting the existence of M2 receptors in these fractions. These data suggest that M2 mAchR are present in all particulate fractions here studied. It is important to emphasize that the M2 muscarinic receptor presents in the light plasma membrane fraction (P1) shows poor selectivity towards the muscarinic antagonists being different from the M2 mAchRs associated with other subcellular fractions isolated from bovine tracheal smooth muscle. PMID- 9334442 TI - Increase of the serotonin metabolite, 5-hydroxyindoleacetic acid, in cerebro spinal fluid and spinal cord of bovines affected with the paraplegic syndrome. AB - The paraplegic syndrome of bovines is a condition characterized by impairment of locomotion, hypoalgesia and finally death within 72 h. The pathogenesis of the syndrome has not been established. In the present work we determined the levels of monoamines and their metabolites in cerebro-spinal fluid and spinal cord of affected animals in order to investigate the functional state of these neurotransmitters. The content of the main metabolite of serotonin, 5 hydroxyindoleacetic acid, was elevated in the cerebro-spinal fluid and in the gray matter of the spinal cord of paraplegic bovines. Serotonin content in the spinal cord did not differ with respect to control animals, but was decreased in the cerebro-spinal fluid of affected animals. Modifications in the noradrenergic system were also observed, but were less consistent, for which reason further studies are needed. These observations indicate an increase in the turnover rate of serotonin in the paraplegic syndrome. The meaning of the described alterations is unknown at the moment. PMID- 9334444 TI - Incorporation of ion channels from the plasma membrane of Leishmania mexicana into planar bilayers. PMID- 9334445 TI - [Role of calcium in the regulation of oxoglutarate oxidation in the rabbit gastric mucosa]. AB - The stimulation of the oxyntic cell by gastric secretagogues is associated to the activation of oxidative metabolism. The mechanisms of this activation are not exactly known. In the present work, we investigated the possible direct effect of Ca2+ on both oxoglutarate oxidation in rabbit gastric glands and oxoglutarate dehydrogenase activity (OGDH) is isolated gastric mitochondria. Both carbachol and Ca2+ ionophores (A23187 and ionomycin) significantly stimulated oxoglutarate oxidation in a dose-dependent manner. This effect was only observed in the presence of low substrate concentrations. BAPTA-AM, an intracellular calcium chelator, significantly inhibited the rate of oxoglutarate oxidation. OGDH activity was stimulated by physiological concentrations of Ca2+ in a dose dependent manner. This effect was reduced by ruthenium red, an inhibitor of mitochondrial Ca2+ uptake, and it was additionally increased by spermine, a stimulant of Ca2+ influx to the mitochondria. The results support the hypothesis that Ca2+ plays a direct role in the activation of oxidative metabolism in the oxyntic cell. PMID- 9334446 TI - Decreased cardiovascular responses to cisterna magna NaCl injection in hypertensive rats. AB - Albino rats were made hypertensive by 1% NaCl in the drinking water for 4-5 months, systolic (SBP) and distolic blood pressure (DBP) were 164.0 +/- 10.1 mm Hg and 118.7 +/- 4.6 mm Hg respectively, vs. control rats whose SBP and DBP were 119.0 +/- 4.4 mm Hg and 86.8 +/- 4.3 mm Hg respectively. Psychosocial stress (1 hour daily for 4-5 months) only raised SBP to 140.0 +/- 5.2 mm Hg; DBP remained unaltered. One percent NaCl intake combined with psychosocial stress, increased SBP and DBP but not significantly beyond the level observed with single 1% NaCl administration. Formerly described control and hypertensive rats were anesthetized with sodium pentobarbital (40 mg/kg) and stereotaxically injected into de cisterna magna (i.c.) with 20 microliters of 1.5 M NaCl solution. During i.c. injection, intraarterial SBP, DBP and heart rate (HR) were continuously recorded. After i.c., 1.5 M NaCl injection, mean arterial pressure (MAP) increased 21.0 +/- 4.0 mm Hg and HR 51.0 +/- 5.0 beats/min in control rats. Rats made hypertensive by 1% NaCl intake showed a significantly lower increase of MAP, 11.0 +/- 1.8 mm Hg; HR increased 37.0 +/- 4.3 beats/min. Rats submitted only to psychosocial stress displayed a response similar to the one described in control rats. Hypertensive rats submitted to both 1% NaCl intake and psychosocial stress had a more intense reduction of the hypertensive and tachycardic response, 8.0 +/ 2.2 mm Hg and 20.0 +/- 3.2 beats/min respectively. Control i.c. injection with the same volume of saline (0.15 M NaCl) did not change significantly SBP, DBP or HR in a separate group of rats. Left ventricle weight (0.754 +/- 0.0333 g) was augmented in the 1% NaCl treated group (0.795 +/- 0.038 g), and increased its protein content by 13.1% (changes not statistically significant). The highest increase of the left ventricle weight (23.7% above control) with no change in its protein content was observed in rats submitted to 1% NaCl intake plus psychosocial stress. In conclusion, chronic high NaCl intake increased blood pressure; psychosocial stress acted as a weak stimulus for SBP and DBP increase, and central nervous system sodium chloride sensitivity for delivering a peripheral sympathetic discharge was found decreased in rats made hypertensive by a high salt intake. PMID- 9334447 TI - [Direct negative inotropic effect of cocaine in rat ventricle strip]. AB - Cocaine, when used as a recreative drug, can induce cardiovascular toxic effects such as acute reduction of left ventricle ejection fraction, which indicates a negative inotropic effect of the drug. The purpose of this study was to clarify the direct negative inotropic effect of cocaine in in vitro conditions. Rat right ventricle strips were incubated in Krebs solution gassed with 95% O2 and 5% CO2 at 37 degrees, and electrically driven with 2 ms square pulses, 17 mA, at 110 systoles/min. Separate experiments were conducted to study cocaine effect at 210 and 310 systoles/min. The contractile force was recorded through a strain-gauge isometric transducer. Cocaine increased contractile force at doses of 0.3-10.0 micrograms/ml, up to 53% over basal contraction. In the presence of 4 x 10(-8) M atenolol, low doses of cocaine did not increase contractile force and at doses between 3.0-10.0 micrograms/ml revealed a depressant activity on heart muscle contractions. Doxazosin (1.0 microM) and yohimbine (0.1 microM) did not modify the positive inotropic effect of cocaine, showing that alpha 1 and alpha 2 adrenergic receptors were not involved in this cocaine ventricle action. Increasing ventricle strip stimulation rate to 210 and 310 systoles/min for 30 seconds, the contractile force was risen by 55% and 95%, respectively. Cocaine at doses 1.0-3.0 micrograms/ml did not modify the physiological increase of contractile force seen upon ventricle rate increase. The mechanism involved in the contractile force increment after ventricle rate increase is a transient rise of cytosolic Ca2+, mainly derived from the sarcoplasmic reticulum and from extracellular fluid. Atenolol (4 x 10(-8) M) exposure of the right ventricle strip intensified the negative inotropic effect of cocaine (3.0-10 micrograms/ml) seen by ventricle stimulation at 210 and 310 systoles/min. The myocardial direct depressant effect of cocaine, in the presence of atenolol, was gradually reversed by extracelular Ca2+ increase at 3.2 and 5.0 mM, respectively. In conclusion, the mechanism of myocardial direct depressant effect of cocaine is related to the beating frequency of the ventricle, which may be associated to interference with the Ca2+ release process from the myocite sarcoplasmic reticulum, and not to calcium entry blockade from extracellular fluid. However, a dpressant effect of cocaine on phase "0" of depolarization, related to its local anesthetic properties can not be ruled out. PMID- 9334448 TI - [Ultrastructural analysis of anastomosis group 9 of Rhizoctonia solani]. AB - The ultrastructure of R. solani AG-9 (S-21, ATCC 62804) was investigated with transmission electron microscopy (TEM). The most important characteristics were those related with cell wall thickness, cytoplasmic matrix composition, number of nuclei and nucleoli and secretory material production. The majority of examined hyphae showed lateral cell walls thinner than those recorded before. The cytoplasmic matrix consistently appeared differentiated into two classes, one formed by a highly electron dense granular fine material and the other one showing a coloidal substance of very low density which give these cells a 'tiger like' aspect. The grannular dense matrix always had abundant free ribosomes and usually surrounded the cytoplasmic organelles and the septal pore apparatus. The somatic cells showed up to 5 nuclei, some of which with three nucleoli. Masses of secretory material surrounded by membrane were regularly seen in the cytoplasm, with sizes similar to those of nuclei. PMID- 9334449 TI - [Analysis of consanguinity in some populations of the V Region, Valparaiso, Chile, from 1880 to 1969]. AB - The population structure of 10 populations ("comunas") in Valparaiso, V Region Chile, was studied through the frequency of consanguineous marriages (%CM) and the coefficient of consanguinity (alpha), in order to know their dynamics, and gather information for clinical and genetic epidemiological studies as well as for isonymy studies. The comunas were grouped according to density: Group I, high density, more than 100 inhabitants/km2; Group II, intermediate, between 25 and 99 inhabitants/km2; and Group III, low, less than 25 inhabitants/km2. Data were obtained from parochial archives and national census, from 1880 to 1969. CM's were divided in: uncle-aunt/nephew-niece (12), first cousins (22), first cousins one removed (23), second cousins (33) and multiple consanguinity (M), and the four subtypes of 12 and 22. Percentage of CM and alpha diminish in time. Groups I and II show similar values, but lower in I, and show a constant decrease. Group III has higher values and considerable fluactuations. Types 12 and 22 contribute mostly to %CM and alpha in the 3 groups. Subtypes of 12 and 22 do not occur at random. This temporary and spatial behavior can be explained because of sociocultural and socioeconomical factors in each group, being density an indicator of endogamy. This behavior is consistent with current coefficients of endogamy obtained by isonymy. PMID- 9334450 TI - [Clinical and hematological changes in calves infected with Anaplasma marginale]. AB - Clinical and hematological changes of six Anaplasma marginale (isolated Zulia) inoculated calves (experimental group) and four healthy calves (control group) were studied during twenty and eighty days before and after infection, respectively. The behavior of the four calves used as control group was stable and no significant changes in the parameters analyzed was observed. The experimental group developed the three typical phases of illness. During the prepatent phase, which lasted a mean of 21.2 +/- 2.56 days, the animals were asymptomatic and no significant changes in the hematological values occurred, but a remarkable transitory decrease in number of lymphocytes from 6.5 x 10(6) to 3.3 x 10(6) cells/ml. The infection during the acute phase produced a highly severe effect in two animals, a severe effect in three animals and a mild effect in one. The effects observed were the following: 1) a fast decrease in haematocrite, ranging from 6 to 10%; 2) values of parasitaemia varied from 15 to 48%; 3) a greater body temperature than the control animals (40.5 vs. 38.5 degrees C); 4) a elevated heart frequency, from 60 to 110 beats/min; 5) an increase in the concentration of neotrophiles from 10 x 10(6) to 13 x 10(6) cells/ml; 6) The number of monocytes also augmented from 3 x 10(6) to 6 x 10(6) cells/ml; and 7) an important decrease of weight gain. The natural course of infection was interrupted with oxytetracycline when the haematocrite of the animal lowered to values less or equal to 10%. Then, the animals showed a rapid recovery with an undetectable parasitaemia and concomitant return to basal line of the rest of the parameters. PMID- 9334451 TI - [Isolation and some properties of the proteinase atroxin from the venom of the snake Bothrops atrox]. AB - A proteolytic enzyme from the venom of Bothrops atrox snake was isolated. It was designed as Atroxin, and three chromatography steps were used to purification: ion exchange chromatography on DEAE-Sephadex A-50 equilibrated with 0.05 M Tris HCl buffer, 1 mM CaCl2 pH 7.4, followed by gel filtration on Sephadex G-50 and Sephadex G-100, respectively, using the same buffer. The enzyme was recovered with a 7.4 folds and 11% of yield. It had a high activity on casein being 7.4 optimus pH. A molecular weight was 19.9 Kd calculated by polyacrilamide gel electrophoresis, and head treatment showed that the enzyme preserves its activity in the range of 37-45 degrees C, while it was decrease when the temperature values were higher. On the other hand, 0.133 mumoles of Ca2+ and Mg2+, and Zn2+ ions (0.266 mumoles) were activators, while EDTA (0.20 mumoles) and sodium azide (0.053 mumoles) were inhibitors. The enzymatic activity was not affected by glicerol (1.33 mumoles) and phenyl methyl sulphonyl fluoride (PSMF) (0.16 mumoles). In addition, iodoacetic acid (0.08 mumoles) was slight inhibitor, but 0.16 mumoles of p-tosyl-1-lysine chloromethyl ketone (TLCK) was activator. Biological assays on mice showed that atroxin produced hemorrhagic and necrosis after 24 h of injection, which was increased by 5 mM calcium chloride. PMID- 9334452 TI - [Systematic quantification of the arteries in lung biopsies of patients with congenital heart defects and its contribution to the therapeutic management]. AB - PURPOSE: To determine the value of the quantitative analysis of lung biopsies from patients with congenital cardiac defects. METHODS: Fourty nine biopsies were examined, from patients: 43 patients increased pulmonary blood flow, 3 with pulmonary atresia and large systemic-to-pulmonary collateral vessels, and 3 with decreased pulmonary flow. The degree of lesion was determined as in Heath-Edwards and of Rabinovitch and col. RESULTS: The Heath and Edwards grade was determined in 41 cases; I in nine; II in 23; III in eight; IV in one; 3 biopsies showed evidence of reduced pulmonary flow and 5 had no signs of vascular disease. The Down patients (7) presented a greater proportion of severe lesions. Quantitative evaluation was obtained in 35 biopsies: 11 had grade B and 24 had grade C. Wall atrophy and dilatation of intraacinar arteries were detected in 7 cases, what suggested the existence of obstructive lesions in proximal vessels, even if not sampled. Medical thickness greater than 2 times the normal were observed in pre acinar arteries from 14 biopsies. CONCLUSION: The morphometric approach allowed us to detect severe lesions which the qualitative analysis alone would not indicate adequately. In the patients presenting decreased pulmonary flow, morphometry made possible to assess if the degree of arterial wall hypertrophy was compatible with a surgery of atrio-pulmonary anastomosis. PMID- 9334453 TI - [Stress test in the elderly. Clinical, hemodynamic, metabolic and electrocardiographic variables]. AB - PURPOSE: To identify in the elderly adaptations imposed by exercise in both sexes. METHODS: 1528 stress tests were performed on subjects divided in: group I (GI) (90%) between 65 to 75 years old, and group II (GII) more than 75 years old. Protocols applied were Bruce (72%), and modified Naughton (28%). Clinical, hemodynamic and electrocardiographic variables were estimated as recommended by the World Health Organization, and the metabolic variables in the adapted Naughton protocols by the American College of Sports Medicine standards. RESULTS: Analysis of GI and GII, respectively disclosed: 1) stress electrocardiogram (ECG): normal, 36 and 35%; ST depression, 20 and 22%; ST elevation, 6 and 1%; ventricular ectopic beats, 11 and 14%; supra ventricular ectopic beats, 5 and 6%; 2) metabolic and hemodynamic variables: the double-product: 26636 (+/-1539) and 23133 (+/-3218) mmHg X bpm (p < 0.0001). Maximum oxygen uptake measured in METS: GI, men, 7.7 (+/-1.9), women 5.4 (+/-0.8) (p < 0.0001); GII, NS, curve of systolic blood pressure: GI, men, 8.4 +/- (0.5), women, 10.6 (+/-1.8) mmHg/Met (p = 0.03); GII- NS. Difference of diastolic blood pressure and heart rate during exercise were similar between the two groups; 3) chest pain was the main clinical variable. CONCLUSION: The more frequent indication for stress testing to evaluate chest pain in GI, did not correspond to a predominance of this symptom in this group, during exercise; in GI, in contrast to what is seen in the young, the curve of systolic blood pressure was greater in women; despite the greater prevalence of coronary artery disease in aged subjects, it was not observed significative differences between the two groups, to ischaemic ST depression. PMID- 9334454 TI - [Evaluation of a cardiac rehabilitation program. Analysis after ten years of follow-up]. AB - PURPOSE: To study the benefits of the cardiac rehabilitation program (CRP) in patients with coronary artery disease (CAD). METHODS: Between 1986 and 1995 we studied 49 patients with CAD, participants of the CRP, 45 (91.83%) of them men. They were compared with a control group of 37 sedentary patients, 33 (89.18%) men. The main parameters analyzed were the duration of exercise, the maximal oxygen consumption (VO2 max), the metabolic equivalent (MET), the functional aerobic impairment (FAI) and the change in the classification of the cardiorespiratory capacity between two graded exercise tests (GTX). RESULTS: There were improvements in all parameters of the GTX analyzed in the two groups. The patients of the CRP presented a better functional capacity than the sedentary patients and, in relation to the duration of exercise, to the VO2 max and to the MET, the differences in the two groups achieved statistical significance (p < 0.05). We did not observe benefits, in relation to the physical conditioning, with a more prolonged permanence of the patients in the program (more than 24 months). There were no cardiovascular complications with the practice of the exercise in the period analyzed. CONCLUSION: The improvement in the duration of exercise, in the VO2 max and in the MET, the more negative variation in the FAI and the improvement in the classification of the cardiorespiratory capacity between the two GTX of the patients of the CRP demonstrate improvement in functional capacity significantly better than sedentary patients. The CRP analyzed was considered a therapeutic method safe and efficient after a coronary event. PMID- 9334456 TI - [Heart surgery in Brazilian Indians]. AB - PURPOSE: Our experience with surgical treatment of heart diseases in Indians living in the Amazon rain forest in primitive stages was reviewed. METHODS: From 1988 to 1995, 18 patients underwent cardiovascular surgical procedures at the Sao Paulo Hospital of the Escola Paulista de Medicina. Seven patients had valvar disease, nine congenital heart defects, one submitral aneurysm and one arrhythmia. Thirteen Indians came from tribes of the Amazon rain forest area: three from the Xavante, two from Waiapi, two from Tucano, two from Macuxi, two from Mayoruna, and one of each tribe of Xikrin, Guajajara, Terena, Surui, Galibi, Cinta-Larga and Pataxo. RESULTS: We performed 22 operations, with two hospital deaths. Follow-up was possible in 87.5% of cases, with one late death. The majority of cases were due to congenital heart defects and in this series it was noted the absence of operations to treat coronary artery disease. The incidence of valve disease was higher in accultured or semi-accultured Indians. CONCLUSION: The surgical treatment of cardiovascular disease has made possible to the surviving indians to return to and be accepted by their fellow tribesmen. PMID- 9334455 TI - [Dietary intervention and plasma cholesterol level in electricity workers]. AB - PURPOSE: To evaluate the efficacy of the diet on plasma cholesterol level above 200 mg/dL and to relate these values to age and occupation at the CPFL (Electricity Supply Company). METHODS: The participants were divided into 3 different groups (A, B and C), according to total cholesterol (TC): < 200 mg/dL (A), > or = 200 mg/dL and < 239 mg/dL (B) and > or = 240 mg/dL (C). Between 1983 and 1991, 688 workers of the company between 21 and 60 years, underwent dosages of TC with diet orientation: those having TC > 200 mg/dL, were identified in groups B and C. In addition to that, the participants were grouped according to their functions at the company. The TC level was dosed by Huang's modified colorimetric method. RESULTS: The mean TC level was 206 mg/dL. The nondiet group presented mean TC of 168 +/- 22.9 mg/dL in 1983 and 192 +/- 27.4 mg/dL in 1991 significantly higher compared to the initial value. Groups B and C, under standard diet, presented mean TC levels of 218 +/- 11.2 mg/dL and 266 +/- 22.5 mg/dL in 1983, respectively, 211 +/- 25.2 mg/dL and 229 +/- 34.4 mg/dL in 1991, with a significant decrease over time (p < 0.0001). In the working status, the managers presented mean TC values significantly higher than other occupations. CONCLUSION: The mean TC level of 206 mg/dL is considered high for Brazilians, and the distribution curve is similar to the American. The response to the standard diet was efficient, decreasing the TC in workers with values > 200 g/dL. Managers presented higher TC levels in relation to other functions, in addition to older age and different job conditions. PMID- 9334457 TI - [Excimer-laser angioplasty for the treatment of restenosis after Wiktor and Gianturco-Roubin stent placement. A new therapeutic alternative]. AB - We report the use of excimer-laser angioplasty for the treatment of Wiktor and Gianturco-Roubin in-stent restenosis of in two patients. Case 1-a 48-year-old man presented unstable angina five months after Wiktor stent was deployed in right coronary artery. Cardiac catheterization revealed stenosis (95%) within the stent. Case 2-a 65-year-old man presented stable angina four months after Gianturco-Roubin stent was deployed in left anterior descending artery. Cardiac catheterization revealed stenosis (80%) within the stent. Excimer-laser angioplasty within the stent reduced the stenosis to 19% and 30%, respectively. The patients recovered and currently, six months post-procedure, are free of chest pain, and cardiac catheterization revealed stenosis to 30% and 35%, respectively, within the stent. Therefore, the procedure was an effective means of treating restenosis after coronary stent placement, and a prospective comparison of excimer-laser angioplasty and other management alternatives to in stent restenosis is needed. PMID- 9334458 TI - [Superior vena cava and right atrium thrombosis successfully treated with streptokinase]. AB - The case of a 56 year-old male with acute lymphoid leukemia and no signs of activity for the last four months is reported. He presented with superior vena cava thrombosis caused by a Hickman catheter, and had positive blood cultures for Candida albicans and Staphylococcus epidermidis. Despite adequate antimicrobial therapy, the fever persisted, and the patient was submitted to surgical thrombectomy. One week following the procedure, the fever returned, and thrombosis of the superior vena cava extending to the right atrium was identified by transesophageal echocardiography (TEE). The patient underwent thrombolytic therapy with streptokinase, and no thrombus could be identified in the control TEE. No hemorrhagic or thromboembolic complication occurred. The patient was discharged with oral anticoagulation. PMID- 9334459 TI - [Ebstein's anomaly in the elderly]. AB - A case of a 62-year-old patient with Ebstein's anomaly is presented. Despite the severe anatomical abnormalities, he was asymptomatic until 61 years of age. Anatomic aspects, clinical features and the diagnostic techniques used are analyzed. PMID- 9334460 TI - [Anatomo-clinical correlation. Case 1/97--Instituto do Coracao do Hospital das Clinicas--FMUSP]. PMID- 9334461 TI - [A step-by-step approach in the diagnosis of neurogenic orthostatic hypotension in the elderly]. PMID- 9334462 TI - [The oxidative hypothesis of atherosclerosis and the use of antioxidants in coronary disease]. PMID- 9334463 TI - [Pravastatin multicenter clinical trial in patients with hypercholesterolemia and multiple risk factors]. AB - PURPOSE: To study whether pravastatin maintain its therapeutic effect when the treatment is indicated to patients with hypercholesterolemy associated to others risk factors. METHODS: In a multicenter investigation 1147 patients, mostly male (609) with a mean age of 52 years were enrolled. The inclusion criteria were cholesterol level above 260 mg/dL or 220 mg/dL if LDL-C greater than 145 mg/dL and one or more risk factors. The patients were treated for 16 weeks, in the first four only with diet orientation and with diet plus 10 mg of pravastatin in the 12 subsequent weeks. RESULTS: Hypertension and smoke were the more frequent risk factors observed in the study population. Only 22% of the patients were on hypolipemiant treatment before enrolling in the study. HDL-C was unchanged and total and LDL-C presented a reduction of approximately 10% with diet orientation. After 10 mg of pravastatin there was a reduction of around 30% of total and LDL-C and a 13% HDL-C elevation. The presence of risk factors did not influence significantly the drug effect, but it is possible to observe that those who smoke and males had lower levels of HDL-C. Obesity and diabetes patients presented greater levels of triglycerides. Therapeutic response to pravastatin is slightly smaller in males and after menopause. CONCLUSION: The number of patients following regular hypolipemiant treatment is still small(22%). Risk factors associated to hypercholesterolemia do not seem to interfere in the lipid lowering response to pravastatin. Pravastatin 10 mg once a day controlled hypercholesterolemia in 85% of the study population. PMID- 9334464 TI - [Prevalence of anal incontinence in the elderly population: an epidemiological study of the elderly population served at the geriatric ambulatory service of the Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo]. AB - The aim of the present work was to assess the prevalence of anal incontinence in the elderly population attended in the "Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo". The method employed was of individual interviews with patient attended in the Geriatric Ambulatory. As part of the assessment patients were classified as to sex, age, color, social-economical class and education, and compared to available data on the elderly population of the State and the City of Sao Paulo. Besides the question on signs and symptoms related to anal incontinence, associated factors such as bowel habit, sensation of incomplete evacuation, urgency to evacuate and soiling were also analysed, as well as, factors possibly related to anal incontinence such as urinary incontinence, use of medication as laxatives, associated diseases, such as neurological affections and diabetes mellitus, number of child births, and assistance received, previous anorectal or perineals operations, and locomotions limiting factor. The statistical method of test of proportion was used to analyse the proportions of the populations interviewed, as to sex and literacy compared to the elderly population of the State and City of Sao Paulo. To analyse the social-economical class of the population interviewed compared to the elderly population economically and non economically active, of the State of Sao Paulo, the test of qui-square adjusted was used. The test of qui-square was used to verify if the population of incontinent and continent, by sex; incontinent and continent in relation to bowel habit, by sex; incontinence sufferer of continuous and descontinuous anal incontinence, by sex; incontinent sufferer of anal incontinence to gas, liquid or solid, and the association of these manifestations, by sex; patients who complained of soiling, by sex; incontinent and continent, previously submitted to anorectal or perineal operations; and continent and incontinent with diabetes mellitus, are equal. To verify the proportions of women who had vaginal delivery in the continent and incontinent group, the test of Mann-Whitney was used. One hundred fourty six patients were interviewed, 43 men of an average 75 years old (62 to 91) and 103 women of an average 72 years old (60 to 88); as to colours, 69.9% were white, 18.5% were mullato, 9.6% were black and 2% were yellow. The comparative analysis with populational data indicated that the population interviewed was representative of the elderly population of the State and the City of Sao Paulo, as to age, color, social-economical class and education. The prevalence of anal incontinence was of 10.9% with no difference between the sexes, intestinal obstipation referred by 15.4% of the men and 28.1% of the women, sensation of incomplete evacuation referred by 21.2% of the patients, and the association of both referred by 13.7% were the most frequently observed functional gastrointestinal alterations. Soiling was referred by 10.3% of the patients with no difference related to sex. The use of medication or laxatives, the presence of diabetes mellitus, neurological affections, previews anorectal or perineal operations, or locomotion limiting factors, have no relation to anal incontinence. The prevalence of urinary incontinence was of 30.1%, and double continence of 6.3% with no statistical difference between sexes. There was statistical evidence of association between childbearing and anal incontinence. PMID- 9334465 TI - [Hematologic aspects of the national hockey team athletes]. AB - Athletes tend to have lower hemoglobin (Hb) concentrations than sedentary counterparts. Sports anemia is used to describe both pseudodilutional anemia and the true anemia of athletes. Pseudodilutional anemia is a beneficial adaptation to endurance training; the two most common causes of true anemia are iron deficiency and intravascular hemolysis. We used questionnaires, physical examination and laboratory investigation to study 19 highly trained hockey athletes and laboratory investigation to study 32 outpatients without hematological diseases. One athlete had anemia (Hb 10.5 g/ dL; 5.44 million red cells/mL; serum iron-13 micrograms/dL; without parasites in the stools); the athletes had Hb = 14.88 +/- 1.33 g/dL and the outpatients had 15.24 +/- 0.74 g/dL; the maximal oxygen uptake of the athletes was 54.0 +/- 6.03 ml/kg/min (112.15 +/- 14.35% of the predicted values). The maximal oxygen uptake of the anemic athlete was 86% of the predicted value. The three athletes with the best maximal uptake had a mean Hb = 15.8 g/dL. In Sao Paulo-Brazil anemia is uncommon among males but elite athletes are in a borderline anemic state, that may impair the physical fitness. PMID- 9334466 TI - [Senile cataract: patient's characteristics and perceptions on a sight restoration communitary project]. AB - A survey was conducted to identify some patient's characteristics and perceptions related to senile cataract and delivery of cataract care. The patients were diagnosed by means of ophthalmic examination carried out on a Rehabilitation of the Aged Campaign at the Hospital das Clinicas of the University of Sao Paulo Medical School. A questionnaire was administered by interviewing 70 subjects, who presented the following characteristics: 32.86% were males and 67.14% were females; 42.86% with ages varying between 50 and 70 years; 67.14% took notice of campaign through TV; 60.00% underwent SUS previous ophthalmic evaluation; 72.86% presented low vision acuity on both eyes for a year or longer, (27.14% for 5 years or longer); 40.00% had previous surgery indication for a year or longer and 80.00% of the patients claimed declared financial reasons to explain their non previous cataract surgery. We recommend eye health educational programs for prevention of blindness and sigh restoration. PMID- 9334467 TI - [Intestinal obstruction caused by inflammatory fibroid polyp. Report of a case]. AB - The inflammatory fibroid polyps are rare lesions of the digestive tract, they were firstly described occurring in the stomach, however their distribution is universal. Several other denominations have been used to same lesion such as fibroma and eosinophilic granuloma. The clinical presentation varies according to the its location, frequently they are mistaken as gastrointestinal neoplasias. Intestinal obstruction is one of the manifestations, mainly when the lesion occurs nearby the ileum cecal region. The authors present a case of intestinal obstruction caused by a inflammatory fibroid polyp located in the ileum cecal valve, treated successfully by right colectomy. PMID- 9334468 TI - [Drug hypersensitivity in AIDS patients: report of a case]. AB - Drug allergy is a common symptom in HIV infected patients, with higher prevalence that in general population. We describe a child with AIDS and adverse reactions to several drugs indicated for Pneumocystis carinii prophylaxis and antibiotics. Reaction to intravenous pentamidine was present in this case and it is not an usual findings in HIV infected children. Alternative therapy and desensitization to TMP-SMX are also discussed. PMID- 9334469 TI - [Pheochromocytoma of the urinary bladder, report of a case and review of the literature]. AB - Pheochromocytoma is a rare neoplasm, found in 0.1% of all hypertensive patients. Extraadrenal pheochromocytomas occur in 18% of all cases and 1% accurate in the bladder. In this study, we report a case of a vesical pheochromocytoma in a 40 year-old male patient with typical clinical symptoms for 6 years. He related episodes of severe headaches and palpitation with increase of the blood pressure after micturition, which decreased within a few seconds and fatigue afterwards. The patient was evaluated through urinary catecholamine (NE: 263 ug/24 h; E: 14 ug/ 24 h; Dopa: 303 ug/24 h; normal range respectively < 80; < 20 and < 400) and plasma catecholamine level determinations before and after micturition (NE: 1.660 ->34.790 pg/ml; E: 55-->231 pg/ml-normal range respectively < 268 and < 75). Magnetic resonance imaging, sonography and 131Iodine-methiliodobenzylguanidine scintigraphy were performed for diagnostic localization. In this case plasma catecholamine level determinations before and after voiding were important to confirm the diagnosis. All imaging techniques were able to disclose the tumor. Typical symptoms, diagnoses and therapy for vesical pheochromocytoma are described and compared to the reports found in the literature. PMID- 9334470 TI - Primary community-acquired pneumonia by Escherichia coli. Case report and review of the literature. AB - A two year old girl with chronic neurologic convulsive disease was admitted with a six day history of pneumonia and, despite treatment, died on hospital day 3. The X-ray revealed right upper lobar pneumonia. The results of pleural effusion and blood cultures drawn on admission yielded a non-typable Escherichia coli. No other source of infection was identified. The authors discuss the clinical and pathophysiological aspects of Escherichia coli pneumonia. PMID- 9334471 TI - [Laparoscopic treatment of duodenal obstruction in a patient with pancreatic cancer]. AB - We present a case report of laparoscopic gastrojejunostomy in a patient with duodenal obstruction from unresectable cancer. We performed an side-to-side intracorporeal gastrojejunostomy using endoscopic stapling devices. The patient had no morbidity and he was discharge on fourth postoperative day. Laparoscopic gastric bypass is an alternative to open procedure in well selected cases. PMID- 9334472 TI - [Educational objectives in medical school: curriculum]. AB - The author discuss in this paper the different options for a curricular reform in medical education presenting the difficulties to implement a curriculum with a broad abrangence. He consider the different possibilities for a curricular structure and evaluated the advantages and disadvantages of each (discipline centered, body-system-based, problem-oriented and clinical presentation curriculum). He argue that most of the critical determinants of physician identity operate not within the formal curriculum but in a more subtle, less officially recognized hidden curriculum. He finish discussing that a significant component of medical training involves the development of a medical morality. PMID- 9334473 TI - [Emergency service in internal medicine-HCFMUSP/Clinical Emergencies Discipline FMUSP: a brief history]. PMID- 9334474 TI - [The Editorial Advisory Committee]. AB - Since 1970, Revista Medica de Chile applies the peer review system as a main step in the selection and improvement of the manuscripts to be published. Over 150 experts participate in this process annually, reviewing up to 5 manuscripts per year. The final decision with regards to to the acceptability of a manuscript remains a responsibility of the Editor. The reviewers are selected by the Editor and his Associates among clinical investigators, prominent subspecialits and basic scientists, according to the nature of the manuscript. Most of them work in Chile. Their names are published and their confidential work is acknowledged in a special chronicle published in the Revista once a year. A small number of these reviewers appears in every issue of the journal identified as Members of its Editorial Advisory Committee. They have been selected by the Editors among those reviewers who deal with a greater number of manuscripts and also those experienced specialists whose opinion is requested when an exceptional conflict of opinions is raised by the authors and their reviewers. After 5 to 10 years of a highly praised collaboration, the previous Committee has been changed and new names were included, starting in this issue of Revista Medica de Chile. PMID- 9334475 TI - [Prognostic value of noninvasive makers of coronary reperfusion in patients with acute myocardial infarction treated with thrombolysis]. AB - BACKGROUND: The immediate prognosis of patients with acute myocardial infarction treated with thrombolysis primarily depends on obtaining a satisfactory coronary reperfusion. AIM: To assess the prognostic power of four markers of coronary artery patency in patients with acute myocardial infarction treated with Streptokinase 1.5 million U within the first six hours of symptoms. PATIENTS AND METHODS: In 807 consecutive patients from the Chilean National Registry of Acute Myocardial Infarction we analyzed the resolution of chest pain and ST segment elevation over 50% within the first 90 min, abrupt CK rise within 8 h and T wave inversion in infarct related EKG leads within the first 24 h after thrombolysis. RESULTS: Global in-hospital mortality was 12.1%. Mortality of patients with the presence of 3 or 4 markers of coronary artery patency was 5.1%, in those with resolution of ST elevation and abrupt CK rise was 6.25% and in those with T wave inversion it was 3.9% (p < 0.001). Multivariate analysis, adjusted by age, gender, risk factors, Killip class and infarct location showed that early T wave inversion was the better predictor of a low in-hospital mortality and that its combination with other markers of coronary artery patency did not increase its prognostic power. Early CK rise and the presence of 3 out of 4 reperfusion criteria were also independent predictors of a low mortality. CONCLUSIONS: Non invasive markers of coronary artery patency are associated with a lower in hospital mortality and may serve as surrogate end points in clinical trials. PMID- 9334477 TI - [Effects of hormone replacement therapy in bone resorption, in post-menopausal women]. AB - BACKGROUND: The effects of different therapies on bone loss rate can be measured using biochemical markers of bone resorption such as urinary hydroxyproline. AIM: To study the effects of hormone replacement therapy on urinary hydroxyproline in postmenopausal women. PATIENTS AND METHODS: Eighty three postmenopausal women without hormone replacement therapy, 54 postmenopausal women receiving hormone replacement therapy and 16 premenopausal women (considered as the control group) were studied. Hydroxyproline was measured in an early morning urine sample, after one day of diet without meat or gelatin. RESULTS: Urinary hydroxyproline in premenopausal women was 33.7 +/- 7.9 mg/g creatinine. The figure for postmenopausal women with hormonal replacement therapy was 33.7 +/- 5.9 mg/g creatinine. Postmenopausal women without replacement therapy had an urinary hydroxyproline of 47.4 +/- 8.5 mg/g creatinine, significantly higher than that of premenopausal and supplemented women. In 21 postmenopausal women, hydroxyproline was measured before and after three months of replacement therapy, values decreased 35.5 +/- 11% in this period and there was a direct correlation between initial values and the degree of reduction (r = 0.69, p < 0.001). CONCLUSIONS: Postmenopausal women receiving hormone replacement therapy have a urinary hydroxyproline excretion similar to that of premenopausal women. PMID- 9334476 TI - [Phenotypic and molecular features of Vibrio cholerae isolated in Chile, Peru and Bolivia. Comparison with environmental reservoirs]. AB - BACKGROUND: Since 1991, a massive cholera epidemic started in Peru and involved most Central and South American Countries. In Chile, 147 cases were registered, the last one in 1995. AIM: To study the phenotypic and molecular features of Vibrio cholerae strains isolated from patients in Peru, Bolivia and Chile and from environmental reservoirs in Santiago, Chile. MATERIALS AND METHODS: The phenotype, biotype and susceptibility to nine antimicrobials was determined for each isolated strain. Also, the genes of cholera and termolabile toxins were determined using DNA probes and a chromosomal restriction profile was done using HindIII, EcoRI and NotI enzymes. RESULTS: Features studied were similar in the 53 strains isolated from patients. Those isolated from environmental reservoirs had different antimicrobial susceptibility, showing ampicillin resistance, and the GT gene was detected in one of 20 strains, compared to clinical samples were it was present in all. Strains isolated from patients and environment had similar chromosomal restriction profiles. CONCLUSIONS: The chromosomal restriction profile gives an image of bacterial genome and it is a useful and reliable tool for the epidemiological surveillance of cholera. PMID- 9334478 TI - [Genetic factors study in family aggregation of schizophrenia in Santiago, Chile]. AB - BACKGROUND: Genetic epidemiological studies indicate that genetic factors contribute to a familial aggregation of schizophrenia. The form of inheritance has not been elucidated but most studies have been done in Caucasian populations. AIM: To study the form of inheritance of schizophrenia in an urban population of Santiago, Chile, containing an admixture of Spanish origin individuals with Southamerican aborigines. SUBJECTS AND METHODS: Forty four randomly selected schizophrenic probands, 22 female, aged 28 to 48 years old, were studied. From them, an extensive genealogical reconstitution was performed. Probands and relatives were interviewed using the structured interview CIDI and DSM-III-R check-list. Schizophrenia was diagnosed using DSM-III-R criteria. Complex segregation analysis was done using Pointer program. RESULTS: The hypothesis of a multifactorial inheritance, without the participation of major genes, could not be rejected. Likewise, the major dominant and co-dominant gene forms of transmission could not be rejected. CONCLUSIONS: Our results show the participation of a major dominant locus and a multifactorial component in the inheritance of schizophrenia, as has been reported elsewhere. PMID- 9334479 TI - [Serum lipid levels in children and teenagers from Concepcion, Chile]. AB - BACKGROUND: There is a relationship between serum lipid levels in children with those of adults. Preventive measures to reduce serum lipid levels should start in childhood. AIM: To study serum lipid levels in a representative sample of children and teenagers from Concepcion, Chile. SUBJECTS AND METHODS: Serum total, HDL cholesterol and triglycerides were measured in 1,286 males and 816 females from 5 to 18 years old in the city of Concepcion. RESULTS: Mean total cholesterol levels were 159 +/- 30 and 162 +/- 31 mg/dl in males and females respectively. The figures for HDL cholesterol were 46 +/- 11 and 47 +/- 11 mg/dl, for LDL cholesterol were 94 +/- 27 and 96 +/- 29 mg/dl and for triglycerides were 80 +/- 35 and 87 +/- 38 mg/dl. Nine percent of males and 12% of females had a total cholesterol over 200 mg/dl. Likewise 10% of males and 11% of females had a LDL cholesterol over 130 mg/dl. CONCLUSIONS: These numbers will help to plan and perform interventions in children, in order to prevent cardiovascular diseases. PMID- 9334480 TI - [Carotid endarterectomy combined with myocardial revascularization: report of 27 patients]. AB - Atherosclerosis is a systemic disease that may involve more than one territory. Myocardial infarction can occur after carotid endarterectomy and stroke is a well documented morbidity of coronary artery bypass grafting. To optimize results, we have performed concomitant carotid endarterectomy and myocardial revascularization in selected cases, with severe disease in both territories. During a 13-year period, 27 patients were submitted for this procedure, 21 (77.8%) were male and the average age was 67.6 years (range 59-81). All patients had high-grade internal carotid artery stenosis, five (18.5%) were symptomatic. Coronary artery disease symptoms were: unstable angina in 12 patients (44.4%) and effort angina in 15 (55.6%). Two patients (7.4%) required reintervention for postoperative bleeding. Two cases (7.4%) had transient renal dysfunction. One patient, with multiple organ failure, died on the 16th postoperative day (3.7%). Follow up was obtained in 26 patients (96.3%). Survival at 5 years was 80.6%, 95.7% of those patients were free of any neurologic symptom. Combined carotid and coronary surgery is a safe treatment option for atherosclerosis of multiple territories in selected patients; long term benefits are also obtained. PMID- 9334481 TI - [Distal renal tubular acidosis and nephrolithiasis in 3 cases of primary Sjogren syndrome]. AB - Tubulo interstitial nephritis, the main manifestation of renal involvement in Sjogren syndrome, may lead to a tubular dysfunction that is usually subclinical. We report three women, aged 32, 35 and 35 years old, with a primary Sjogren syndrome and symptomatic type I or distal tubular acidosis. Two patients had nephrolithiasis and one a nephrocalcinosis. Two had a basal hyperchloremic metabolic acidosis. The ammonium chloride acidification test was abnormal in all, demonstrating a distal tubular defect. None had proximal tubular dysfunction. All had an urinary pH over 6.5 and hypocitraturia and none had hypercalciuria. Renal calculi were composed of calcium oxalate and calcium phosphate in two patients and calcium phosphate and ammonium phosphate in the other. All women had positive antinuclear antibodies with mottled pattern, two had anti Ro antibodies and positive rheumatoid factor and one had hypergammaglobulinemia. None had anti La antibodies, crioglobulinemia or monoclonal proteins. PMID- 9334482 TI - [Lead poisoning: clinical picture, diagnosis and related environmental factors]. AB - BACKGROUND: The predisposing environmental factors and clinical picture of lead poisoning, are not well known. AIM: To describe the clinical picture, diagnosis and treatment of lead poisoning in individuals under industry exposure. SUBJECTS AND METHODS: Twenty of 38 workers of a printing press were studied. Their medical and labor histories were recorded. Lead in air in the working area and the dose received by the workers was measured. RESULTS: The environmental lead in the linotype room was 25% over the accepted values. Twelve of the 20 workers were poisoned and three were highly exposed. Poisoned workers were working in areas with high environmental lead concentrations, were in direct contact with the metal, had plasma lead concentrations over 70 micrograms/dl and an average exposure time of over 17 years. Their clinical picture was not specific and related to plasma and environmental lead concentrations, length of exposure and type of contact. CONCLUSIONS: Lead poisoning must be suspected among workers exposed to high environmental concentrations and its treatment consists in withdrawing poisoned subjects from polluted areas. PMID- 9334483 TI - [Anatomo-radiological study of the cranio-cervical region in 60 students from Universidad de la Frontera, Chile]. AB - BACKGROUND: Twenty to thirty percent of the population has craniomandibular anomalies, that are closely related to craniocervical disorders. AIM: To evaluate the craniocervical region from a radiological point of view in healthy young adults. SUBJECTS AND METHODS: A lateral head and neck radiological study, using the technique described by Rocabado, was done to 60 Chilean young adults, aged 19 to 24 years old. RESULTS: The cervical curvature was altered in 70% of subjects (kyphosis in 35%, straightening in 33.3% and lordosis in 1.7%). Sixty eight percent had alteration of the hyoid triangle (in the plane in 31.7% and inverted in 37%). The distance between CO and C1 was altered in 48% (less than 4 mm in 15% and more than 9 mm in 33%). An altered posteroinferior angle was observed in 40% (less than 96 degrees in 30% and over 106 degrees in 10%). CONCLUSION: A high percentage of alterations of the craniocervical region was detected in healthy adults. PMID- 9334484 TI - [Hepatic veno-occlusive disease associated to the use of azathioprine in a renal transplant recipient]. AB - We report a 30 years old male, recipient of a kidney allograft and treated with azathioprine, who eighteen days after transplantation had a clinically asymptomatic elevation of total bilirubin and alkaline phosphatases. Nineteen months later, he presented with mild ascites, with a total bilirubin of 3.5 mg/dl, alkaline phosphatases of 308 U/L (normal < 170 U/L) and a prothrombin time at 55% of control. A liver biopsy showed sinusoidal and perivenular fibrosis without inflammation, compatible with chronic venous obstruction. Hepatic veno occlusive disease is an infrequent complication of azathioprine use. PMID- 9334485 TI - [Pulmonary edema associated with the ingestion of hydrochlorothiazide]. AB - Hydrochlorotiazide may cause pulmonary dema. We report two female patients aged 58 and 69 years old, who after the ingestion of this diuretic, were admitted to an intensive care unit with fever, digestive symptoms, severe dyspnea, hypotension and a hemodynamic pattern resembling hypovolemia. Both had a good response to volume repletion and vasoactive drugs. Despite the severity of the clinical picture, they had a full and fast recovery. This type of reaction to hydrochlorotiazide is probably idiosyncratic and must be considered in the differential diagnosis of pulmonary edema. PMID- 9334486 TI - [Endoscopic therapy in advanced pancreatic cancer. Minimally invasive surgery]. AB - We report a 70 years old male with an advanced pancreatic carcinoma who was admitted to the hospital due to vomiting, progressive jaundice and severe itching. An upper gastrointestinal endoscopy showed an extrinsic duodenal compression and the abdominal ultrasound, a dilated biliary tract. During an endoscopic retrograde pancreatography a papilotomy was performed and a 7 F biliary stent was installed. Also, an endoscopic percutaneous gastrostomy was done, installing a jejunostomy. These procedures allowed a better quality of life in this patient. PMID- 9334488 TI - [Cutaneous metastases. Clinical pathological review]. AB - Cutaneous metastases may be the first clinical manifestation of a visceral carcinoma, its incidence fluctuates between 0.7 and 9% of the carcinomas and normally indicate a dismal prognosis. The most frequent morphologies of cutaneous metastases are metastatic nodules, sclerotic forms that appear as infiltrated and stiff plates, the inflammatory variety that resembles an erysipelatous plate, mammary and extra-mammary Paget's disease that appear as erythematous erosive and crusty plates resembling dermatitis. PMID- 9334487 TI - [Cerebral hemorrhage associated with the use of phenylpropanolamine. Clinical cases]. AB - We report three women aged 33, 63 and 43 years old receiving an anorexigenic medication that contained phenylpropanolamine who, 30 min to 3 h after the ingestion of this medication, presented a frontal hematoma, a left putaminal hematoma and a subaracnoidal hemorrhage, respectively. Cerebral angiography, performed in all of them, did not show any vascular abnormality. This complication of phenylpropanolamine use, has been reported previously abroad and the mechanism of production, could be related to an elevation of blood pressure and the production of a vasculitis by the drug. The risk of this complication must be taken into account when prescribing phenylpropanolamine. PMID- 9334489 TI - [The crisis of internal medicine]. AB - The rapid technological progress in medicine has lead to a better physiopathologic and etiological knowledge of adult diseases, producing a fragmentation of internal medicine and a great development of its subspecialties. Furthermore, specialization grants a professional, scientific and academic status. Contrarywise technological enrichment has impoverished the human relationship of clinical practice. The training at the Medical School has a great responsibility towards General Internal Medicine, and the technical capacity of teachers should be revised. The practical tutorial teaching at the hospital should be accomplished by Internists with a general clinical orientation. Outpatient clinics should be incorporated as places where a more clinical and less technological medicine could be taught. Internship should be a period in which General Internal Medicine should be valorized, continuing in post-graduate training and continuous teaching. General Internal Medicine is going through a crisis; Medical Schools and the Medical society have the responsibility to overcome it. PMID- 9334490 TI - [The need for nuclear medicine specialists and gamma cameras]. AB - BACKGROUND: The ideal number of nuclear medicine specialists in relation to the needs of a given population should be defined by the institutions training such specialists. AIM: To establish the relationship between the number of gammacameras, nuclear medicine specialists and the number of inhabitants in different countries, including Chile. MATERIAL AND METHODS: Information was obtained from the International Atomic Energy Agency, Britannic and Canadian Nuclear Medicine Societies, Pan-American Health Organization and American College of Nuclear Physicians. RESULTS: Industrialized countries have approximately 1 specialist and 0.5-3.3 gammacameras per 100,000 inhabitants. The respective figures in Chile are 0.25 and 0.25. Due to the unequal distribution of resources in Chile, the specialist and gammacameras rate in the Metropolitan area is 0.6, some regions or communities do not have these resources and in other communities the rate can raise up to nine. CONCLUSIONS: Compared to industrialized countries, Chile lacks nuclear medicine specialists and gammacameras, and these resources are unevenly distributed. PMID- 9334492 TI - [Hypothyroidism and autoimmune chronic thyroiditis in Copiapo, Chile]. PMID- 9334491 TI - [Diagnostic value of cerebrospinal fluid adenosine deaminase determination]. PMID- 9334493 TI - [The origin of the University of Chile's Clinical Hospital "Jose Joaquin Aguirre"]. PMID- 9334494 TI - Anatomy of the cranioencephalic structures of the goat (Capra hircus L.) by imaging techniques: a computerized tomographic study. AB - A topographic study of the cranioencephalic structure was carried out by computerized tomography on Canarian breed adult goats of medium size and weight, with similar cephalic parameters. In this way, transversal, sagittal and horizontal tomographic images were obtained. Identification of the observed anatomic structures represents the basis of this work from which applicable specie data are derived. PMID- 9334495 TI - Vimentin- and desmin-positive cells in the moulting budgerigar (Melopsittacus undulatus) skin. AB - The distribution of vimentin- and desmin-positive cells in the budgerigar (Melopsittacus undulatus) dermis was investigated by means of immunohistochemical reactivity with the commercially available (Euro-Diagnostics) polyclonal antibodies. The staining pattern for vimentin in the paraffin sections was generally comparable to that in other animal species with regard to endothelial cells, vascular wall cells, muscle cells and fibroblasts. The modified Schwann cells in the inner core of the Herbst corpuscles reacted distinctly with anti vimentin and anti-desmin. Some connective tissue cells in the superficial dermal layer, in the feather papilla and along the pulp core inside of the regenerating feathers were particularly well stained with anti-vimentin. Fibroblast-like cells of the regenerating feathers, particularly at the base of the pulp, also reacted strongly with anti-desmin. The findings were discussed with regard to references. PMID- 9334496 TI - The soft-tissue components of the vomeronasal organ in pigs, cows and horses. AB - The soft-tissue components of the vomeronasal organ of the pig, the cow and the horse were studied with the aid of dissection, microdissection, and light microscopy and immunohistochemistry of series of transverse sections. In horses, the rostral end of the incisive duct was blind: thus, unlike in pigs and cows, there was no communication between the vomeronasal organ and the oral cavity. In all three species, the central part of the vomeronasal duct bore the 'typical' respiratory/ receptor epithelium lining on its lateral and medical walls. The rostral part of the duct was characterized by stratified columnar epithelium, while more caudal parts bore simple columnar type. The patterns of distribution of glands, blood vessels and nerves were closely associated with the patterns of distribution of duct linings. The distribution of soft-tissue components in pigs was less clearly defined than in cows and horses. Of the three species, nerves were detected in the rostral half of the vomeronasal parenchyma only in the horse. PMID- 9334497 TI - Development of myelination in the bovine fetal brain: an immunohistochemical study. AB - In this study, the development of myelination in the fetal bovine brain (age range: between 1-2 and 8-9 months) was examined applying: 1. Immunohistochemical staining methods and antibodies against bovine proteolipid protein (PLP), synthetic tridecapeptide of bovine PLP, human myelin basic protein (MBP), human myelin-associated glycoprotein (MAG); and 2. Using the Luxol fast-blue (LFB) technique. PMID- 9334498 TI - The developmental relation among mesonephros, gonad and external genitalia in the fetus of goat (Capra prisca). AB - The mesonephros, the gonad, the external genitalia and the developmental relationships between them were examined on 21 goat fetuses, 29 to 92 days old. Three fetuses were not sexually differentiated, however 11 were males and 7 females. The development of the gonad could be divided in a not differentiated stage and a gonadal and external genitalia differentiation stage. A close relationship between the fetal gonad and the mesonephros was found during the stage of differentiation. The epicenter of this relation is the giant mesonephric nephron, a peculiar structure of mesonephros, from which epithelial cells presumably migrate into the gonad. PMID- 9334499 TI - A quantitative study of the aorta of the chicken (Gallus domesticus). AB - The different segments of the aorta were studied quantitatively in five age groups of chicken. It was observed that the widest diameter and the thickest media occur on the 56th day in each segment, except for the thoracic aorta, which had the widest diameter on the 42nd day and the thickest media on the 70th day. The percentage of the intima, media and the adventitia, as compared with the total wall thickness, was determined and it was found that this value was quite similar in age group, but the differences were significant from one segment to another. The ratio between the total wall thickness and the lumen diameter was also calculated in each segment. PMID- 9334500 TI - Study on the developmental changes in angulation of spinal nerves, the length of the dorsal roots and spinal nerves in sheep. AB - This study reports the angulation of spinal nerves, the length of dorsal roots, the length of spinal nerves and the transverse and vertical diameters of the spinal cord during pre- and postnatal life in sheep of the Mehraban breed in Iran. The spinal cord of these animals was divided into 4 regions with respect to the angulation of spinal nerves. The first region was from the first to fifth cervical and the second was from the sixth cervical to the eighth thoracic in fetuses and from the six cervical to fourth thoracic in adults. The third region was from the ninth thoracic to second lumbar in fetuses and from the fifth thoracic to second lumbar in adults, and the fourth region was from third lumbar to fourth sacral in the animals of all age groups. The length of dorsal roots from their point of emergence from the spinal cord to the dorsal root ganglia showed a direct correlation with the length of the spinal nerves. While angulation of the spinal nerves showed a converse correlation with the length of either spinal nerves or dorsal roots. The transverse diameters of the spinal cord were always longer than the vertical diameters. The greatest diameters (vertical 7.00 mm; transverse-10.00 mm) are recorded at C1, T1 and L6 in adult sheep. PMID- 9334501 TI - Ultrastructure of the zymogenic-cell lineage in abomasal mucosa of adult cattle. AB - The ultrastructural differentiation and maturation of the neck cells and the zymogenic cells during physiological cell renewal were investigated in the abomasal oxyntic-gland region of cattle. Immature neck cells of the distal isthmus and proximal neck exhibit transitional morphology to the predominantly mucous isthmus cells. Neck cells confined to the glandular neck are characterized by bipartite peptic-cored mucous secretory granules. In a proximal-distal gradient along the neck, a progressive increase in the peptic granular component and concomitant reduction in mucous components paralleled by proliferation of the rough endoplasmic reticulum creates pre-zymogenic cells in the proximal glandular base. These, in turn, give rise to mature zymogenic cells with pure peptic secretory granules and typical zymogenic cell morphology. In the depth of the gland, older degenerative zymogenic cells are found. Variations in size and number of the zymogenic granules point to different secretory activities of the mature zymogenic-cell population of the glandular base. These results favour the conception of a zymogenic-cell lineage arising within the isthmus and passing through different developmental stages, including neck cells, during their migration down the gland. PMID- 9334502 TI - Basic and lectin histochemical characterization of bovine gustatory (von Ebner's) glands. AB - The Bovine tongue possesses numerous circumvallate papillae (8-16 each side). The troughs around the papillae are the openings of the ducts of the gustatory (von Ebner's) glands. In this study, we have characterized in situ the glycosidic composition of the secretion of bovine gustatory glands using traditional histochemical methods and lectin histochemistry with and without prior neuraminidase (sialidase) digestion. The lectin-horseradish peroxidase conjugates employed were: PNA, DBA, SBA, WGA, LTA, UEA I and ConA. Acinar cells show a diffuse positivity towards PAS and Alcian blue at pH 2.5 and the most intense and homogeneous lectin staining was obtained with PNA. This indicates that bovine gustatory glands secrete glycoproteins with 1,2-glycol containing hexoses and carboxyl-rich glycoconjugates and that galactosyl (beta 1-->3) Nacetylgalactosamine is the most frequent sugar residue present in these glycoproteins. Results were compared with data reported in the literature on the same glands of other species. PMID- 9334503 TI - Gross anatomy of the portal vein and its tributaries in the opossum (Didelphis albiventris). AB - The gross anatomy of the portal vein (V. portae) and its tributaries was studied through anatomical methods, i.e. dissection, corrosion and diaphanization, in 45 opossums (Didelphis albiventris). In all animals the portal vein was formed by the junction of the cranial mesenteric, caudal mesenteric and lienal veins (V. mesenterica cranialis, V. mesenterica caudalis and V. lienalis, respectively). Many collateral tributaries were observed running into the portal venous trunk. PMID- 9334504 TI - Morphological study of bovine pregnant corpus luteum cells. AB - The bovine pregnant corpus luteum was examined morphologically, to clarify the appearance and properties of the intramitochondrial bodies (IMB) in mitochondria of the large luteum cell (LLC). The incidence and diameter of the IMB (200-900 nm) showed a tendency to increase with the advance of pregnancy. Histochemically, the IMB reacted positively with protein, and immunohistochemically, they reacted IMB with 11 alpha-hydroxyprogesterone-1-11 succinyl-bovine serum albumin (P-BSA). The IMB seemed to combine with protein, and these granules did not undergo exocytosis. The cytoplasm of LLC contained another type of small electron-dense granules that were 150-200 nm in diameter, had an orifice for secretion, and reacted with relaxin. Small luteum cells (SLC) revealed crystalline-like inclusions. It is suggested that these inclusion cells participate in the synthesis of steroids. PMID- 9334505 TI - The Institute of Living: 175 years of innovation and excellence. PMID- 9334506 TI - The Hartford Retreat for the Insane: an early example of the use of "moral treatment" in America. PMID- 9334507 TI - Primum non nocere: somatic treatments for mental illness in the early 20th century. PMID- 9334508 TI - Dynamic therapy in the decade of the brain. AB - Data from diverse sources suggest that psychotherapy works by changing the functioning of the brain. This observation is part of a larger phenomenon involving neural plasticity. An extraordinary amount of research is demonstrating that neither gene expression nor the anatomy or physiology of the brain is static. Environment and genetics are in an ongoing interaction with one another, and this mutual influence informs our latest understanding of how psychotherapy appears to affect brain functioning. Research on disorders such as schizophrenia, obsessive-compulsive disorder, cancer, and panic disorder provide convincing data in this regard. For a full understanding of the etiology and pathogenesis of mental illness, as well as its treatment, the psychodynamic dimension of meaning must be incorporated. PMID- 9334509 TI - Alzheimer's disease: cognitive and behavioral pharmacotherapy. AB - Alzheimer's disease is a progressive degenerative disease with cognitive and behavioral manifestations. The pathophysiology of Alzheimer's disease is increasingly well understood, leading to approved and experimental therapies. Mutations on chromosomes 1, 14, and 21 can cause the disease and are sometimes present in patients with early onset Alzheimer's disease. Older patients- comprising the majority of Alzheimer's disease victims--have a variety of risk factors including age, gender, history of head trauma, low education level, and apolipoprotein genotype. The pathogenetic process leads to regional cell loss and biochemical deficits. The mutations and risk factors lead to increased amyloid production or accumulation and nerve cell death. Neurons atrophy in the cerebral cortex and in source nuclei of important neurotransmitters. The deficiency of acetylcholine can be partially remedied by cholinesterase inhibitors that produce modest cognitive and behavioral improvement. Anti-amyloid agents, antioxidants, anti-inflammatory drugs, estrogens, and calcium channel blockers may all slow the progression of Alzheimer's disease by interfering with specific steps within the disease cascade. Neuropsychiatric symptoms are also common in Alzheimer's disease and may be ameliorated by conventional psychotropic agents. Clinicians can currently offer substantial help to Alzheimer's disease patients, and the future promises more effective pharmacotherapy. PMID- 9334511 TI - Assessing outcomes: identifying psychiatric patients with severe and persistent illness. AB - BACKGROUND: Identification of psychiatric patients with severe and persisting impairments can facilitate treatment, aid in program planning, and provide data for cost-of-care projections. METHODS: In this prospective study of patient outcomes, 1,679 inpatients were classified on admission using a functional status measure developed by the authors. Consenting subjects were reassessed at discharge and at 3, 6, and 12 months postdischarge to determine what proportion of patients classified as low functioning on admission remained so at follow-up. RESULTS: Patients classified as low functioning on admission represented 23.4% of the sample; the proportion that remained low functioning at the follow-ups ranged from 56.1% to 65.2%. Compared to the high functioning group, three times more low functioning patients were rehospitalized within 12 months of discharge (9.4% vs 32%). CONCLUSIONS: Patients with increased risk of persisting disability can be identified on admission using commonly available clinical measures. Of patients with low functioning on admission, more than half will have long-term impairment. PMID- 9334510 TI - Physician compliance with practice guidelines. AB - BACKGROUND: Physician adherence to established practice standards has become an important national issue. Despite the proliferation of formal standards of practice, there is little evidence that the mere availability of guidelines results in changes in physician practices. This paper presents the results of a study of the effectiveness of a computerized monitoring and notification system in increasing physician compliance with treatment guidelines. METHODS: This study prospectively compared medical staff practices in two one-year periods utilizing a computer system which tabulated noncompliance and provided reminders. RESULTS: Overall, there was a statistically-significant decrease in the number of alerts issued in year two compared to year one; alerts were issued on 15% vs 29% of all patients (P < .001). The average number of alerts per patient decreased to .20 from .41. CONCLUSIONS: The study results indicate that a clinical decision support system such as that described can improve adherence to treatment guidelines. PMID- 9334512 TI - Alcohol screening and brief intervention: where research meets practice. AB - Numerous studies have consistently shown that quick screening instruments can identify people whose drinking is likely to present health risks and that low cost, brief interventions are effective in reducing drinking among many such at risk drinkers. This article describes the results of a one-year policy analysis that explored how alcohol screening and brief intervention (SBI) can be moved to widespread clinical applications in the United States. It introduces the concept of risky drinking and considers the potential of this new technology to reduce it. The research evidence behind this approach is reviewed, and a description of current programs in this and other countries beginning to apply SBI is provided. Economic issues attendant to applications are identified and discussed. The potential for applications in health care is analyzed and summary conclusions from market research are set forth. Recommendations are offered for immediate action. PMID- 9334513 TI - Alcohol and drug abuse among Connecticut youth: implications for adolescent medicine and public health. AB - In 1995, a statewide survey of alcohol and other drug use was conducted in a random sample of approximately 4,000 7th to 12th graders in public schools in Connecticut. The survey, part of a statewide substance abuse treatment needs assessment, showed that use of tobacco, alcohol, and marijuana was widespread and increasing, particularly among younger students. Connecticut's students reported higher rates of substance use compared to their peers nationwide. Substance use differed according to age, gender, ethnic background, and community type. It was estimated that almost 1-in-10 senior high school students should receive a diagnostic evaluation for substance abuse, with half likely to need a treatment referral. Most of these adolescents had not received treatment for their substance abuse. Primary-care physicians can play a key role in reducing adolescent substance abuse through prevention messages, screening for drug use, brief interventions, and timely referrals to appropriate intervention services. PMID- 9334514 TI - New directions in clinical psychopharmacology. AB - This paper reviews the current clinical use of psychotropic drugs and suggests new directions for future drug development. The author reviews the contemporary pharmacotherapy of depression, anxiety disorders, schizophrenia, and bipolar disorder and summarizes current trends in the drug treatment of psychiatric aspects of HIV disease. The increasing use of combination pharmacotherapy is discussed, as well as the impact of pharmacokinetics in clinical psychopharmacology. The use of botanical products for psychiatric disorders and tobacco use in the psychiatric patient are also addressed. Current research is producing drugs that have greater specificity in their mechanism of action effecting faster clinical response with minimum adverse effects. PMID- 9334515 TI - Neuropsychological rehabilitation in the treatment of schizophrenia. AB - The Schizophrenia Rehabilitation Center at the Institute of Living has begun to develop rehabilitation techniques based on a neuropsychological approach to ameliorate the cognitive deficits associated with schizophrenia. A study was designed to test the hypothesis that patients receiving attention skills training would demonstrate improved performance on neuropsychological tests measuring concentration. Five out of the seven subjects significantly improved on more than half of the neuropsychological measures. In the two case studies reviewed, significant increases in test scores were observed more frequently during cognitive rehabilitation then when psychoeducation classes were substituted for the attention skills training. In addition, patients showed improvement on the Positive and Negative Syndrome Scale and Quality of Life Scale. The data support the efficacy of attention skills training using a process specific approach. However, it is uncertain if attention skills rehabilitation is a necessary or sufficient factor in producing the therapeutic change. PMID- 9334516 TI - Risk management and treatment of sexual disinhibition in geriatric patients. AB - BACKGROUND: Upwards of 7% of cognitively impaired elderly are reported to exhibit sexually disinhibited behaviors. These behaviors may be the result of either a chronic history of sexual disinhibition, regression, or sequel to a stroke, surgical intervention, vascular insult, blow to the head, or cardiac event in which observed cognitive deterioration is the acute symptom. Elderly patients who are sexually disinhibited may exhibit a behavior that makes it difficult to manage them at home or in a nursing home. METHODS: A review of the treatment of sexual disinhibition with neurohormones is presented; guidelines for assessing risk and risk management are proposed; and a five-year study with 39 geriatric out patients with cognitive impairment and sexual disinhibitions reviewed. Case examples of sexual aggressives are followed by treatment recommendations in which an algorithm is presented. RESULTS: The treatment algorithm recommends beginning selective serotonin reuptake inhibitors medication before considering estrogen (preferably a patch) or antiandrogen therapy. The estrogen patch led to excellent treatment results in elderly demented men with sexual disinhibition. CONCLUSIONS: Elderly demented patients who are sexually disinhibited may be managed successfully with neurohormones if SSRI medication proves unsuccessful. PMID- 9334517 TI - Informed consumer participation in managed Medicare. PMID- 9334518 TI - Psychotherapy and psychiatry: a partnership for the future. PMID- 9334519 TI - Psychiatry in Israel: towards the year 2000. PMID- 9334520 TI - Methylphenidate: between rhetoric and data. PMID- 9334521 TI - Hey! It was just a joke! Understanding propositions and propositional attitudes by normally developing children and children with autism. AB - Two- and three-year-old children were asked why a speaker named objects falsely. Most produced explanations in terms of the speaker's intention to joke. This implies a sensitivity to two distinct levels in language: the proposition itself and the propositional attitude. Children with learning difficulties showed a similar competence. In contrast, most children with autism failed to explain such false statements in these terms, instead merely describing them as "wrong." This was not simply due to a metalinguistic deficit, as they correctly answered questions about what a speaker had said the object was. These results suggest the normally developing toddler has a remarkable facility in processing propositional attitudes, while children with autism do not; and that such an ability is broadly independent of general intelligence. PMID- 9334522 TI - Psychometric properties of the K-SADS-PL in an Israeli adolescent clinical population. AB - The main objective of this study was to evaluate the reliability and validity of the Schedule for Affective-Disorders and Schizophrenia for School-Aged Children Lifetime Version (K-SADS-PL) in an Israeli sample. Fifty-seven adolescent admissions to two psychiatric units were evaluated with the Hebrew translation of the K-SADS-PL. Inter-rater and mother-child agreement were evaluated for diagnoses, and 31 affective symptoms. K-SADS diagnoses were also compared with two-month unit evaluation diagnoses as a measure of consensual validity. The reliability of assessing psychosocial functioning was also examined. Reliability and validity of diagnoses were good to excellent. Mother-child agreement fared less well on all measures. The use of semi-structured interviews and DSM-IV criteria in an Israeli adolescent psychiatric setting are discussed. PMID- 9334523 TI - Depressive disorder in pre-adolescence: comorbidity or different clinical subtypes? (A pharmacological contribution). AB - The study consisted of evaluating 80 out-patients for DSM-IV diagnoses and giving 75 mg trazodone for four months to children aged 9-13 who fulfilled DSM-IV criteria for Major Depressive Disorder (n = 22); Major Depressive Disorder and Generalized Anxiety Disorder (n = 24), Major Depressive Disorder and Learning Disorder NOS (n = 24); Major Depressive Disorder and Oppositional Defiant Disorder (n = 10). They were followed up weekly by a psychiatrist blind to the original evaluations and to the precise purpose of the study. Trazodone emerges as safe and effective for over 50% of the sample. Those with depression alone and with LD NOS did particularly well, and those with ODD did particularly badly. The presence of psychopathology in the parents was associated with poorer response to trazodone, and life events were also associated with a poor response. PMID- 9334524 TI - Psychiatric in-patient treatment for children and adolescents in the UK: criteria for admission. AB - There are few published data concerning the clinical criteria that determine admission of children or adolescents to psychiatric in-patient units. This paper reports a case-notes study of the criteria employed by UK clinicians, prior to the introduction of a purchaser-provider health care market into the UK National Health Service. Two child and adolescent psychiatrists rated the case notes of 24 in-patients, and 72 out-patients, matched for age and sex, according to symptomatology, social adversity and severity of behavioral and emotional impairment. The findings indicate that social adversity does not predict admission. Certain symptoms such as sexualized or severe antisocial behavior may even act to diminish the likelihood of admission. However, the principal finding is that, within a non-market, free health care system, the degree of clinical impairment suffered by the child or young person appears to be a prime determinant of the decision to admit. PMID- 9334525 TI - The effect of methylphenidate on the cognitive and personality functioning of ADHD children. AB - Twenty Israeli children diagnosed as having Attention Deficit Hyperactivity Disorder (ADHD) participated in the study. The study used a double-blind, cross over, placebo-control design, in which each child received a three-week treatment of methylphenidate (MPH), and a three-week treatment with a placebo. Their teachers completed the Conners' Teacher Rating Scale (CTRS) along with ratings of other variables prior to the beginning of the study and during the two treatment periods. The subjects were tested on a specially designed battery of both cognitive and personality tests at the end of each treatment period. The results showed significant differences between ratings of children's behavior in pretest and subsequent placebo and MPH treatment periods. Comparison of the MPH and placebo treatment revealed no differences in any of the measures assessing cognitive ability and personality characteristics. PMID- 9334526 TI - Formal thought disorder in offspring of schizophrenic parents. AB - OBJECTIVE: We examined potential early markers of schizophrenia using measures of formal thought disorder in offspring of parents with schizophrenia, other mental illness and no mental illness. METHODS: Two blind raters coded formal thought disorder in adolescent/early adult offspring of 42 schizophrenic, 39 other mental illness, and 36 no mental illness parents. In addition to parental diagnosis, we compared the individual offspring diagnosis with severity of formal thought disorder. Within the schizophrenia, other mental illness and no mental illness offspring groups, we examined the relationship between severity of formal thought disorder and performance on cognitive and motor tasks. RESULTS: There were no statistically significant differences in formal thought disorder by parent or offspring diagnoses. Within the offspring group of parents with schizophrenia, the subjects with higher formal thought disorder scores performed significantly worse on the cognitive battery than those with lower formal thought disorder scores. Offspring of the other mental illness group with higher formal thought disorder scores, however, showed more deficits on motor tasks than those with lower formal thought disorder scores. CONCLUSION: Formal thought disorder may reflect underlying cognitive dysfunction in the offspring of parents with schizophrenia. Motor dysfunction in the offspring of parents with other psychiatric illness might be associated with formal thought disorder. PMID- 9334527 TI - A comparative study of defense styles of bulimic, anorexic and normal females. AB - We have compared the defense styles of anorexic (N = 41), bulimic (N = 37) and normal females (N = 72) living in Paris, using the Bond Defense Style Questionnaire relating to 17 defenses: sublimation, anticipation, suppression, undoing, idealization, reaction formation, projection, passive aggression, acting out, isolation, devaluation, autistic fantasy, denial, displacement, dissociation, splitting and somatization. The objective of this study was to better understand the personality structure in terms of psychological defenses of adolescent girls and young women suffering from anorexia or bulimia nervosa (DSM III-R). Data showed significant differences of psychological functioning between control subjects and eating disorder subjects, particularly for the projection, undoing and sublimation defenses. Anorexics differed from the bulimic females only on the passive aggression, isolation and devaluation defenses. These data are discussed in relation to the hypothesis that anorexics and bulimics can be situated on the same continuum ranging from normal to eating disorders with certain common psychological features as risk factors shared by the eating disorder females. PMID- 9334528 TI - A general hospital study of attempted suicide in adolescence: age and method of attempt. AB - Attempted suicide in adolescence is a major health problem and recent reports indicate a dramatic increase in the frequency of attempted and completed suicide among adolescents. The aim of this study was to evaluate the characteristics of adolescent admissions with attempted suicide in a large general hospital. The files of all children and adolescents admitted due to a suicide attempt between the years 1984 and 1994 were examined retrospectively, with regard to age, sex, method of attempt, season, year and length of hospitalization. Four hundred and four admissions with attempted suicide were recorded. The majority (83.7%) were females and drug overdose was the most common (92.8%) method used. Thirty patients (7.5%) repeated the attempt during the study period. No specific time of the year was associated with an increase in adolescent suicide attempt admissions. The most remarkable finding was that younger adolescents had a higher probability of performing a violent suicide attempt. Hypotheses for these trends are examined together with possible ramifications for treatment provision. PMID- 9334529 TI - Psychogenic cough tic: a case report. AB - A case of psychogenic cough tic is presented to demonstrate the problems and complications that can arise from misdiagnosis of this dramatic and disabling disorder. It can be confused with TS and may be treated more aggressively than is beneficial. A single case with a recurrent course is described with the hypothesis of a possible autoimmune contribution. The combination of organic precipitants and emotional maintaining factors suggests that psychogenic cough tic is a condition that lies between somatization and transient tic disorder. PMID- 9334530 TI - Developmental disorder, Tourette disorder and schizophrenia: a case study. AB - We present a unique case of an 18-year-old male who had a classic picture of schizophrenia preceded by a well documented history of Tourette Disorder and a developmental disorder. The subject, a member of an ongoing study on first admission psychosis, has been systematically evaluated and followed up for two years, and the interesting neuropsychological findings are presented and compared to those of the rest of the sample with a diagnosis of schizophrenia. The triad of schizophrenia, Tourette Disorder and developmental disorder is described for the first time in a subject with an adult type schizophrenia. Possible neurodevelopmental impairments explaining the clinical picture are discussed in view of the recent literature. PMID- 9334531 TI - Risperidone-induced nocturnal enuresis. PMID- 9334532 TI - Seniors Outreach Program: an alternative service for older people with substance use problems. PMID- 9334533 TI - Community-based hypertension control programs that work. AB - Hypertension is the number one public health problem in the United States, particularly among African Americans. Although the National High Blood Pressure Education Program, started in 1972, led the way to a substantial decrease in morbidity and mortality from this disease, the percentage of African American hypertensives whose conditions are detected, treated, and controlled continues to lag behind that of white hypertensives. Community-based programs at locations where people congregate-for example, churches, barbershops, beauty salons, firehouses, housing projects, and worksites-can play a valuable role in increasing the number of African American hypertensives who receive treatment. Physicians can be a potent force for the development of these programs by acting as consultants to define the scope and function of lay volunteers and by promoting these programs in a variety of other ways. PMID- 9334534 TI - Investigating the relationship between economic status and HIV risk. AB - Although anecdotal information suggests that there is a relationship between socioeconomic status and HIV risk, there have been few investigations of that possible relationship. Understanding that relationship can have important implications for designing and implementing prevention programs. This study investigated the relationship between indicators of socioeconomic status and HIV prevalence in Massachusetts using seroprevalence data from publicly funded test sites. HIV seroprevalence was found to differ depending on demographic groups and the health care insurance/provider. Those who had no insurance or were Medicaid recipients had higher rates of HIV infection. Homeless individuals were also at higher risk. Further, low-income ZIP codes in Massachusetts were four times more likely to have high seroprevalence rates among residents voluntarily testing for HIV. Thus, HIV seroprevalence appears to be associated with socioeconomic status in this group of voluntarily tested individuals. PMID- 9334535 TI - Strategies for suppression, containment, and eradication of resurgent tuberculosis. AB - This review provides strategies for the suppression, containment, and eventual eradication of resurgent tuberculosis. Some ethnic minority communities are at greatest risk because of the prevalence of human immunodeficiency virus/acquired immunodeficiency syndrome, poverty and malnutrition, congregate living situations, aberrant lifestyles, illegal immigration, and underemployment among these populations. Proposed strategies include the education of the population at risk as well as health care providers to permit the optimization of preventive, diagnostic, and therapeutic technologies. Also necessary is the development of effective, safe, newer medications to enhance patient compliance and decrease drug resistance. Strategic planning embraces national socioeconomic policy to permit adequate resources to combat poverty and malnutrition, to rebuild the infrastructure of the public health system, and to improve access to health care among rural and urban dwellers. It is concluded that these efforts must continue to ensure the eradication of tuberculosis. PMID- 9334536 TI - Factors associated with return visits to a homeless clinic. AB - Associations between characteristics of homeless clients and their return visits to a nurse-managed primary health care clinic were examined using a retrospective chart review of 1,467 records from clients seen between 1991 and 1994. Client characteristics examined included age, education, race, gender, sheltered status, report of chronic disease, and report of family living in the area. Only 47 percent of clients made return visits to the clinic. Logistic regression indicated that those with reported chronic disease, males, whites, and those living on the street were more likely to have returned to the clinic for care than those without chronic illness, females, nonwhites, and those living in some type of shelter. Results suggest the need for program planning and evaluation for this population, which particularly considers women, nonwhites, and those without chronic disease as target groups for services. PMID- 9334537 TI - An analysis of unmet need for HIV services: The Houston Study. AB - HIV/AIDS is indicative of general institutional neglect that disproportionately affects minorities, poor, and underserved populations. Among women and minorities, HIV infection is associated with preexisting economic distress. Moreover, socioeconomic resources, gender, and race/ethnicity may determine access to medical and nonmedical services that affect disease progression. An analysis of data collected for the Ryan White Care Act needs assessment in Houston, Texas, was performed to assess the effects of socioeconomic and demographic factors on unmet needs for existing medical, social, and counseling services, adjusting for the effects of illness and substance abuse. Results indicated that lower income and Hispanic ethnicity were associated with the unmet need for medical services. Higher income was positively associated and African American ethnicity was negatively associated with the unmet need for social services. Also, higher income and private insurance were negatively associated with counseling services. The authors suggest that these latter findings may result from program eligibility requirements and respondents' hierarchy of needs, respectively. PMID- 9334538 TI - Atypical bipolar symptoms. PMID- 9334539 TI - Infection-triggered OCD. PMID- 9334540 TI - The play therapy controversy. PMID- 9334541 TI - Does methylphenidate influence cognitive performance? PMID- 9334542 TI - ADHD and dangerous driving. PMID- 9334543 TI - Prevalence of depression in Europe. PMID- 9334544 TI - SSRI treatment of enuresis. PMID- 9334545 TI - Somatoform disorders in children and adolescents: a review of the past 10 years. AB - OBJECTIVE: To review the literature on somatoform disorders in children and adolescents relevant to recertification by the American Board of Psychiatry and Neurology. METHOD: The psychiatric, pediatric, and psychological literatures were searched for clinical or research articles in the past 10 years dealing with somatization and somatoform disorders. RESULTS: Somatizing presentations are organized conceptually; somatization disorder, body dysmorphic disorder, hypochondriasis, conversion disorder, vocal cord dysfunction, pain disorder, and recurrent abdominal pain are described in children and adolescents; empirical evidence for treatment efficacy is scant, but clinically reasonable approaches are applied. CONCLUSION: More developmentally appropriate diagnostic schemas and better outcome studies are needed in all the somatoform disorders for children and adolescents. PMID- 9334546 TI - Child and adolescent psychiatry residency training: current issues and controversies. AB - OBJECTIVE: To examine major current influences on child and adolescent psychiatry (CAP) residency training, highlighting the most common problems. Potential solutions and unresolved dilemmas are presented. METHOD: Data were gathered from empirical studies, review articles, national census data, and discussions over the past decade at national meetings on recruitment of residents and faculty; clinical, didactic, and research training; the impact of managed care and changes in graduate medical education funding; and the professional development of CAP residents. RESULTS: Overall there are significant problems recruiting U.S. medical students and attracting faculty into CAP training programs. Economic forces, including decreased reimbursements from managed care and the federal government, are threatening the survival and vitality of training programs. Managed care is harmful for sound residency training and identity formation of the child and adolescent psychiatrist. CONCLUSIONS: The integrity of CAP residency training in the future will depend on increased efforts of teaching hospitals and programs to develop fiscally viable systems of care integrated with residency training; seek new sources of training subsidies; modify traditional models of clinical and didactic curricula; and foster greater collaboration between training programs locally and nationally. PMID- 9334547 TI - Parent management training: evidence, outcomes, and issues. AB - OBJECTIVE: To describe and evaluate parent management training (PMT) as a treatment technique for oppositional, aggressive, and antisocial behavior. METHOD: Recent research is reviewed on the efficacy of PMT; factors that contribute to treatment outcome; the range of outcomes related to child, parents, and family; and variations of treatment currently in use. Limitations are also discussed related to the impact of treatment, clinical application, and dissemination of treatment. RESULTS AND CONCLUSIONS: PMT is one of the more well investigated treatment techniques for children and adolescents. Notwithstanding the large number of controlled studies attesting to its efficacy, fundamental questions remain about the magnitude, scope, and durability of impact. PMID- 9334548 TI - Impact of family type and family quality on child behavior problems: a longitudinal study. AB - OBJECTIVE: In the context of substantial changes in family types and even family quality in recent times, this study is concerned with the extent to which family type and quality impacts on child behavior problems. METHOD: A sample of 8,556 pregnant women were enrolled in a prospective, longitudinal study. Details of changes in family type and family quality (assessed using Spanier Dyadic Adjustment Scale) were used to predict three second-order syndromes developed from the Child Behavior Checklist and administered to the mothers when the child was 5 years of age. RESULTS: Mothers who experienced no partner changes (married and single) reported the lowest rates of child behavior problems for the three syndromes used in this study. In addition, mothers who more often described their relationship with their partner as poor also reported the highest rate of child behavior problems across all three syndromes. Adjustment for possible confounders did not alter these findings. CONCLUSION: Both changes of partner and dyadic conflict appear to lead to child behavior problems, with the latter factor appearing to have a greater impact than the former. Mothers who experienced no partner changes and no conflict appeared to have children with the fewest behavior problems. PMID- 9334549 TI - Mothers' and fathers' reports of children's reactions to naturalistic marital conflict. AB - OBJECTIVE: To examine how children, aged 8 to 11 years, react to observing their parents' naturally occurring marital conflict in their own homes and to compare reactions among children raised in homes characterized by different types of marital conflict. METHOD: For 5 1/2 weeks, mothers and fathers of 110 children used daily home diaries to record independently their children's reactions to marital conflict. Logistic regression compared the likelihood of exhibiting specific reactions to conflict among children from marriages characterized by low conflict (LOWCON; n = 37), nonphysical conflict (NOPHYCON; n = 35), and physical conflict (PHYCON; n = 38). RESULTS: Mothers' and fathers' reports indicate that children from PHYCON homes were more likely than children from LOWCON and NOPHYCON homes to leave the room, misbehave or appear angry, and appear sad or frightened. CONCLUSIONS: These results support the notion that different histories of exposure to marital conflict may contribute to the way in which children react to their parents' marital conflict. PMID- 9334550 TI - Case study: ego-dystonic anger attacks in mothers of young children. AB - Parental anger can have detrimental effects on children and can contribute to physical abuse. Ego-dystonic anger attacks are an underrecognized psychiatric symptom that occurs in associated with depression, with other psychiatric disorders, and in the absence of comorbid disorders. They are characterized by overwhelming anger and autonomic arousal occurring upon provocation viewed as trivial by the individual, and they respond well to treatment with serotonergic antidepressants. Consequently, they represent a readily treatable problem. Four cases of anger attacks in mothers of young children are described to illustrate the importance of recognizing and treating anger attacks. PMID- 9334551 TI - Attention-deficit hyperactivity disorder with bipolar disorder: a familial subtype? AB - OBJECTIVE: To clarify the nosological status of children with attention-deficit hyperactivity disorder (ADHD) who also satisfy diagnostic criteria for bipolar disorder (BPD). METHOD: Blind raters and structured psychiatric interviews were used to examine 140 children with ADHD, a sample of 120 non-ADHD comparisons, and their 822 first-degree relatives. Data analyses tested specific hypotheses about the familial relationship between ADHD and BPD. RESULTS: After stratifying the ADHD sample into those with and without BPD, the authors found that (1) relatives of both ADHD subgroups were at significantly greater risk for ADHD than relatives of non-ADHD controls; (2) the two subgroups did not differ significantly from one another in their relatives' risk for ADHD; (3) a fivefold elevated risk for BPD was observed among relatives when the proband child had BPD but not when the proband had ADHD alone; (4) an elevated risk for major depression with severe impairment was found for relatives of ADHD + BPD probands; (5) both ADHD and BPD occurred in the same relatives more often than expected by chance alone; and (6) there was a trend for random mating between ADHD parents and those with mania. CONCLUSIONS: The data suggest that comorbid ADHD with BPD is familially distinct from other forms of ADHD and may be related to what others have termed childhood onset BPD. PMID- 9334553 TI - Cerebral glucose metabolism in adolescent girls with attention deficit/hyperactivity disorder. AB - OBJECTIVE: Low cerebral metabolic rates for glucose (CMRglc) have been reported in a small sample of girls with attention-deficit/hyperactivity disorder (ADHD). This study was an effort to replicate this finding in a larger independent sample. METHOD: Using positron emission tomography and [18F]fluorodeoxyglucose, CMRglc were compared between 10 girls with ADHD (14.10 +/- 1.91 years) and 11 normal girls (14.3 +/- 1.70 years). RESULTS: Global CMRglc was similar between ADHD and control girls. Lateralization of normalized CMRglc differed significantly between ADHD and control girls in parietal and subcortical regions, with rCMRglc lower on the left than on the right side in girls with ADHD, and conversely in control girls. The sylvian area of the parietal region and the anterior putamen of the subcortical region were the main contributors to this effect. Normalized rCMRglc of the hippocampus was higher in ADHD than in control girls. Sexual maturation was the only clinical characteristic that differed between present and previous samples, and it correlated with global CMRglc. CONCLUSIONS: Although failing to confirm abnormally low CMRglc in girls with ADHD, this study suggested that (1) functional interactions between sex and brain development may contribute to ADHD pathophysiology, and (2) sexual maturation should be controlled in future CMRglc studies of adolescent girls. PMID- 9334552 TI - Serotonin, aggression, and parental psychopathology in children with attention deficit hyperactivity disorder. AB - OBJECTIVE: To explore the relationship between central serotonergic (5-HT) function and history of parental aggression in aggressive and nonaggressive boys with attention-deficit hyperactivity disorder (ADHD). METHOD: History of psychiatric symptoms was assessed in the biological parents of 41 boys with ADHD. The relationship between 5-HT function in aggressive and nonaggressive probands, as assessed via the prolactin response to fenfluramine (FEN) challenge, and parental history of aggression was examined. RESULTS: Aggressive boys with a parental history of aggressive behavior had a significantly lower prolactin response to FEN challenge than aggressive boys without a parental history of aggression. Nonaggressive boys had a prolactin response midway between those of the two aggressive subgroups, and their prolactin response did not vary as a function of parental aggression. Children subdivided on the basis of parental history of other psychiatric symptoms did not differ in their response to the FEN challenge. CONCLUSIONS: These data indicate an association between parent aggressive behavior and lower 5-HT function in aggressive boys with ADHD but do not indicate the extent to which this association is environmentally and/or genetically transmitted. There may be different neurochemical mechanisms in familial and nonfamilial aggressive children, which have clinical implications for pharmacological interventions. PMID- 9334554 TI - Effects of methylphenidate on preschool children with ADHD: cognitive and behavioral functions. AB - OBJECTIVE: To report on implications for methylphenidate treatment of this very young age group and the need to examine factors related to achieving compliance. METHOD: Thirty-one children with attention-deficit hyperactivity disorder (ADHD), aged 4 to 6 years, participated in a double-blind, placebo-controlled study using placebo, 0.3 mg/kg, and 0.5 mg/kg of methylphenidate twice per day. RESULTS: Improvements related to medication were obtained on cognitive tests of attention and impulsivity as well as behaviors assessed by parent rating scales. In an interactive setting with their mothers, attentional abilities and the children's ability to work more productively also showed improvement. However, no changes were obtained with respect to the children's tendency to comply with parental requests. Side effects increased slightly with the high dosage of medication but remained mild. CONCLUSION: The results suggest that methylphenidate can be used to improve the functioning of preschool-age children with ADHD, in a manner similar to their school-age counterparts. PMID- 9334555 TI - Religiosity and depression: ten-year follow-up of depressed mothers and offspring. AB - OBJECTIVE: This study examines maternal religiosity as a protective factor against depression in offspring. METHOD: Sixty mothers and 151 offspring were independently assessed over the course of a 10-year follow-up. Maternal and offspring religiosity were assessed on the basis of self-report of the importance of religion, the frequency of attendance of religious services, and religious denomination. Depression was assessed using the Schedule for Affective Disorders Lifetime version. Maternal bonding style was assessed through offspring report on the Parental Bonding Instrument. A series of logistic regressions were run to predict offspring depression status, taking into account maternal religiosity, offspring religiosity, and mother-offspring concordance of religiosity. RESULTS: Maternal religiosity and mother-offspring concordance of religiosity were shown to be protective against offspring depression, independent of maternal parental bonding, maternal social functioning, and maternal demographics. CONCLUSION: Maternal religiosity and offspring concordance with it may protect against depression in offspring. PMID- 9334556 TI - Dysthymic disorder in clinically referred preschool children. AB - OBJECTIVE: This clinical and descriptive study examined the existence, phenomenology, and frequency of dysthymic disorder in a sample of clinically referred preschool children. In addition, the frequency of DSM-IV symptoms and the alternative research criterion for dysthymic disorder were investigated. METHOD: Three hundred consecutive preschool admissions (aged 2 to 6 years) to a child development unit were given a comprehensive evaluation by a treatment team. Data were collected from multiple informants based on the suitability of each source. RESULTS: The findings indicated that eight children met criteria for dysthymic disorder according to the DSM-IV criteria and the alternative research criterion for dysthymic disorder. CONCLUSIONS: The results support the existence of dysthymic disorder in preschool-age children. Recommendations are made for future versions of DSM as well as the appropriateness and significance of various sources of information, such as the child, parents, teachers, and clinician observations, for the evaluation of symptoms of dysthymic disorder in preschoolers. PMID- 9334557 TI - Prediction of positive outcomes for adolescent psychiatric inpatients. AB - OBJECTIVE: To identify individual, parent/family, and treatment follow-through predictors of outcome for adolescent psychiatric inpatients 6 months after hospital discharge. METHOD: Eighty-nine adolescents participated in a comprehensive baseline evaluation during psychiatric hospitalization. Baseline measures included the Diagnostic Interview Schedule for Children, Social Adjustment Inventory for Children and Adolescents, Reynolds Adolescent Depression Scale (RADS), and Suicidal Ideation Questionnaire-Junior (SIQ-Jr). Structured telephone follow-up interviews assessed treatment follow-through, suicidal behaviors, rehospitalizations, living changes, and social adaptive functioning. The RADS and SIQ-Jr were also readministered. RESULTS: Baseline indices of adolescent functioning emerged as the strongest predictors of outcomes. Hierarchical multiple regression analyses indicated that baseline depression severity, a cluster of parent/family indices, and medication follow-through were significant predictors of outcome depression severity. Baseline social adaptive functioning, presence/absence of conduct disorder, and medication follow-through were significant predictors of outcome social adaptive functioning. CONCLUSIONS: The nature and course of adolescent psychopathology was difficult to disrupt, with baseline characteristics as the strongest predictors of outcome. Nevertheless, the significance of medication follow-through as a predictor suggests that treatment-related gains are possible. PMID- 9334558 TI - Paroxetine in children with major depressive disorder: an open trial. AB - OBJECTIVE: To study the clinical benefits and safety of paroxetine (PXT) in a group of patients younger than 14 years old, with a diagnosis of major depressive disorder. METHOD: Patients were recruited from a psychiatric outpatient clinic. They were assessed for disease severity using the Clinical Global Severity (CGS) scale at 1 month, 3 months, and 8 months. Intensity of the therapeutic response (the reduction in CGS) was measured after 1 and 3 months. All patients were given open-label treatment with PXT. RESULTS: At the start of treatment, most patients had "severe" depression (mean CGS score, 3.02). This improved to a score of 2.2 after 1 month and to 1.22 after 3 months of treatment. All patients experienced a complete remission of symptoms by the end of treatment (mean 8.4 months). PXT was well tolerated. Only 4 of the 45 patients experienced (mainly gastrointestinal) side effects; in 3 patients the side effects were of mild severity, and none required withdrawal from the trial. CONCLUSIONS: PXT can be effective in treating major depression and was well tolerated by children younger than 14 years old. This result indicates that PXT should be investigated further in children, in double-blind trials. PMID- 9334559 TI - A longitudinal twin study of temperament and behavior problems: common genetic or environmental influences? AB - OBJECTIVES: To assess the longitudinal covariance between emotionality, activity, and sociability (EAS) temperamental traits and anxious/depressed behavior, attention problems, delinquent behavior, and aggressive behavior and to assess the significance of genetic and common environmental influences on the temperament and behavior relations. METHOD: Parental responses to the Child Behavior Checklist and the EAS Temperament Survey were collected from five national cohorts of Norwegian same-sex twins. The final sample consisted of 759 twin pairs aged 7 through 17 at 2-year follow-up. RESULTS: High emotionality predicted anxious/depressed behavior, attention problems, delinquent behavior, and aggressive behavior. The influence on delinquent and aggressive behavior was stronger in boys. Aggressive behavior was further predicted by high activity scores, especially in younger children. Significant genetic influence was found for the covariance between emotionality and attention problems and emotionality and aggressive behavior. CONCLUSION: Emotionality was the strongest temperamental predictor of behavior problems. The mechanisms involved in the associations between temperament and behavior problems appeared to differ with kind of behavior problems. PMID- 9334560 TI - Low resting heart rate at age 3 years predisposes to aggression at age 11 years: evidence from the Mauritius Child Health Project. AB - OBJECTIVE: Previous studies indicate that low resting heart rate is probably the best-replicated biological correlate of childhood antisocial and aggressive behavior. Nevertheless, there have been few longitudinal tests of this relationship, little control over potential confounds and mediators, and no test of its cross-cultural generalizability. This study tests the hypothesis that low resting heart rate at age 3 years predicts aggression at age 11 years. METHOD: Resting heart rate at age 3 years was assessed in 1,795 male and female children from Mauritius. Aggressive and nonaggressive forms of antisocial behavior were assessed at age 11 years using the Child Behavior Checklist. RESULTS: Aggressive children had lower heart rates than nonaggressive children (p < .001). Conversely, those with low heart rates were more aggressive than those with high heart rates (p < .003). There were no interactions with gender or ethnicity. Evidence was found for specificity of low heart rate to aggressive forms of antisocial behavior. Group differences in heart rate were not attributable to 11 biological, psychological, and psychiatric mediators and confounds. CONCLUSIONS: It is concluded that low resting heart rate, a partly heritable trait reflecting fearlessness and stimulation-seeking, is an important, diagnostically specific, well-replicated, early biological marker for later aggressive behavior. PMID- 9334561 TI - Verbal dichotic listening in boys at risk for behavior disorders. AB - OBJECTIVE: The association between deficits in verbal processing skills and disruptive psychopathology remains one of the most frequently replicated findings in all of child psychiatry. This study uses a dichotic consonant-vowel listening test to examine the potential neural basis for this association. METHOD: A series of 87 young boys recruited from a sample at risk for disruptive disorders received standardized psychiatric, neuropsychological, and language skills assessments. Approximately 1 year later, these boys received a reassessment of their psychiatric status and a test that assesses the neural basis of language processing ability, a dichotic consonant-vowel listening test. RESULTS: Disruptive psychopathology predicted reduced right ear accuracy for dichotic syllables, indicative of a deficit in left hemisphere processing ability. Deficits in reading and language ability also correlated with right ear accuracy for dichotic syllables. CONCLUSIONS: Boys with disruptive behavior disorders, relative to at-risk but nondisruptive boys, exhibit a deficit in verbal processing abilities on dichotic listening tasks. This deficit in verbal processing ability is also manifested as low scores on standardized tests of reading achievement and language comprehension. PMID- 9334562 TI - Summary of the practice parameters for the assessment and treatment of children and adolescents with conduct disorder. AB - This summary of the practice parameters describes the diagnosis, treatment, and prevention of conduct disorder in children and adolescents. The rationales for these recommendations are based on a review of the scientific literature and clinical consensus, which are contained in the complete document. Clinical features of youths with conduct disorder include predominance in males, low socioeconomic status, and familial aggregation. Important continuities to oppositional defiant disorder and antisocial personality disorder have been documented. Extensive comorbidity, especially with other externalizing disorders, depression, and substance abuse, has been documented and has significance for prognosis. Clinically significant subtypes exist according to age of onset, overt or covert conduct problems, and levels of restraint exhibited under stress. To be effective, treatment must be multimodal, address multiple foci, and continue over extensive periods of time. Early treatment and prevention seem to be more effective than later intervention. PMID- 9334563 TI - Introduction: history and development of the practice parameters. PMID- 9334564 TI - Practice parameters for the psychiatric assessment of infants and toddlers (0-36 months). American Academy of Child and Adolescent Psychiatry. AB - These practice parameters describe the psychiatric assessment of infants and toddlers (0-36 months) and support the growth of infant and toddler psychiatry, a rapidly developing field. Infants and toddlers are brought to clinical attention because of concerns about emotional, behavioral, relational, or developmental difficulties. It is axiomatic that the infant or toddler must be understood, evaluated, and treated within the context of the family. A perspective that is developmental, relational, and multidimensional and that borrows from the knowledge of multiple disciplines is essential. Collaborative efforts support the urgent need and incomparable opportunity to understand and to intervene early and preventively with young children and their families. PMID- 9334565 TI - Practice parameters for child custody evaluation. American Academy of Child and Adolescent Psychiatry. AB - These practice parameters are presented as a guide for clinicians evaluating the often delicate and complex issues surrounding a child custody dispute. The historical basis of child custody and the various judicial presumptions that have guided courts are reviewed. The differences between performing child custody evaluation and engaging in traditional clinical practice are emphasized. Issues that are common to all child custody disputes are presented, including continuity and quality of attachments, preference, parental alienation, special needs of children, education, gender issues, sibling relationships, parents' physical and mental health, parents' work schedules, parents' finances, styles of parenting and discipline, conflict resolution, social support systems, cultural and ethnic issues, ethics and values, and religion. In addition, special issues that complicate custody evaluations are discussed, including infants in custody disputes, homosexual parents, grandparents' rights, parental kidnaping, relocation problems, allegations of sexual abuse, and advances in reproductive technology, such as frozen embryos, oocyte donation, and artificial insemination. An outline is provided that describes the complete evaluation process, from assessing referrals and planning a strategy through conducting clinical interviews, writing the report, and testifying in court. PMID- 9334566 TI - Practice parameters for the assessment and treatment of children and adolescents with anxiety disorders. American Academy of Child and Adolescent Psychiatry. AB - Anxiety disorders comprise one of the most prevalent categories of psychopathology in children and adolescents. These revised practice parameters highlight the DSM-IV changes for anxiety disorders and review the literature related to the assessment and treatment of anxiety disorders in children and adolescents. Up-to-date information on longitudinal outcome data, assessment of anxiety, parent-child interventions, and use of selective serotonin reuptake inhibitors has been added to the previous parameters, published in September 1993. Recommendations for evaluation and multimodal approaches to treatment are presented. PMID- 9334567 TI - Practice parameters for the assessment and treatment of children, adolescents, and adults with attention-deficit/hyperactivity disorder. American Academy of Child and Adolescent Psychiatry. AB - These practice parameters review the literature on children, adolescents, and adults with attention-deficit/hyperactivity disorder (ADHD). There are three types of ADHD: predominantly inattentive, predominantly hyperactive-impulsive, and combined. Together, they occur in as many as 10% of boys and 5% of girls of elementary school age. Prevalence declines with age, although up to 65% of hyperactive children are still symptomatic as adults. Frequency in adults is estimated to be 2% to 7%. Assessment includes clinical interviews and standardized rating scales from parents and teachers. Testing of intelligence and academic achievement usually are required. Comorbidity is common. The cornerstones of treatment are support and education of parents, appropriate school placement, and pharmacology. The primary medications are psychostimulants, but antidepressants and alpha-adrenergic agonists are used in special circumstances. Other treatments such as behavior modification, school consultation, family therapy, and group therapy address remaining symptoms. PMID- 9334568 TI - Practice parameters for the assessment and treatment of children and adolescents with conduct disorder. American Academy of Child and Adolescent Psychiatry. AB - These practice parameters address the diagnosis, treatment, and prevention of conduct disorder in children and adolescents. Voluminous literature addresses the problem from a developmental, epidemiological, and criminological perspective. Properly designed treatment outcome studies of modern psychiatric modalities are rare. Ethnic issues are mentioned but not fully addressed from a clinical perspective. Clinical features of youth with conduct disorder include predominance in males, low socioeconomic status, and familial aggregation. Important continuities to oppositional defiant disorder and antisocial personality disorder have been documented. Extensive comorbidity, especially with other externalizing disorders, depression, and substance abuse, has been documented and has significance for prognosis. Clinically significant subtypes exist according to age of onset, overt or covert conduct problems, and levels of restraint exhibited under stress. To be effective, treatment must be multimodal, address multiple foci, and continue over extensive periods of time. Early treatment and prevention seem to be more effective than later intervention. PMID- 9334569 TI - Practice parameters for the assessment and treatment of children and adolescents with substance use disorders. American Academy of Child and Adolescent Psychiatry. AB - These practice parameters describe the assessment and treatment of children and adolescents with substance use disorders and are based on scientific evidence regarding diagnosis and effective treatment as well as on the current state of clinical practice. Given the paucity of research on the treatment of substance use disorders in children and adolescents, many of the recommendations are drawn from the adult literature and current clinical practice. These parameters consider risk factors for substance use and related problems, normative use of substances by adolescents, the comorbidity of substance use disorders with other psychiatric disorders, and treatment settings and modalities. PMID- 9334570 TI - Atmospheric mold spore counts in relation to meteorological parameters. AB - Fungal spore counts of Cladosporium, Alternaria, and Epicoccum were studied during 8 years in Denver, Colorado. Fungal spore counts were obtained daily during the pollinating season by a Rotorod sampler. Weather data were obtained from the National Climatic Data Center. Daily averages of temperature, relative humidity, daily precipitation, barometric pressure, and wind speed were studied. A time series analysis was performed on the data to mathematically model the spore counts in relation to weather parameters. Using SAS PROC ARIMA software, a regression analysis was performed, regressing the spore counts on the weather variables assuming an autoregressive moving average (ARMA) error structure. Cladosporium was found to be positively correlated (P < 0.02) with average daily temperature, relative humidity, and negatively correlated with precipitation. Alternaria and Epicoccum did not show increased predictability with weather variables. A mathematical model was derived for Cladosporium spore counts using the annual seasonal cycle and significant weather variables. The model for Alternaria and Epicoccum incorporated the annual seasonal cycle. Fungal spore counts can be modeled by time series analysis and related to meteorological parameters controlling for seasonallity; this modeling can provide estimates of exposure to fungal aeroallergens. PMID- 9334571 TI - Effect of ambient temperature and energy demands on digestive functions in leaf eared mice (Phyllotis darwini) from central Chile. AB - The leaf-eared mouse, Phyllotis darwini, is a nocturnal rodent inhabiting the semiarid and Mediterranean habitats of northern and central Chile. Previous observations suggested that in the field, individuals may change food intake according to seasonal changes in ambient temperature. We therefore anticipated that P. darwini should increase food intake in response to lower ambient temperature. As predicted, results of food trials and digestive measurements demonstrated that P. darwini increases food intake and assimilation at lower ambient temperatures but does not increase food mean retention time. At lower ambient temperatures, individuals increase digestive tract size thus improving body mass maintenance and perhaps survival during winter. PMID- 9334572 TI - Climatic controls of the cool human thermal sensation in a summertime onshore wind. AB - Afternoon observations in summer comparing shoreline with inland atmospheric conditions were made during onshore winds at Victoria, British Columbia, Canada. The onshore wind came from a cool water surface. Mean monthly water temperatures near to shore were between 11 and 11.5 degrees C. The onshore wind brought lower air, ground surface radiant and sky radiant temperatures; lower humidity and greater wind speed. All of these combine to produce a cooler human environment at the shoreline than inland. The relative importance of climatic elements in producing the cooler environment was assessed using sensitivity analyses with eight different human thermal exchange models/indices. Air temperature and wind speed had the greatest effect, followed by ground surface radiant temperature, sky radiant temperature and humidity. Wind speed is the most practical element to consider when trying to maximize human comfort along the shoreline. PMID- 9334573 TI - Heat stress in Greece. AB - For 12 selected synoptic stations of the Greek Weather Service, the daily 12 UTC values of the thermal index Predicted Mean Vote (PMV) were calculated for the years 1980 to 1989. The locally varied occurrence of diverse thermal sensation and particularly of strong heat stress were analysed in relation to the human biometeorological significance. With the help of a statistical model, PMV values of individual stations were transformed into a high-resolution bioclimatic map. The map presents the average annual number of days with at least strong heat stress (PMV > 3.0). PMID- 9334574 TI - A study of nephron function in normal tropical residents using the creatinine and lithium clearances. AB - The kidney bears the brunt of the demands of a tropical climate for water and electrolyte homeostasis. We hypothesised that a tropical climate may cause adaptive changes in the entire organism leading to altered renal function in our subjects. Hence renal function data for residents of a temperate climate may not be applicable to tropical residents. We therefore sought to elucidate renal function in subjects residing in a tropical climate. We used lithium clearance, CLi, a non-invasive tool for assessing proximal tubular function in humans, and endogenous creatinine clearance, CCr, to estimate proximal tubular function and glomerular function, respectively, in our subjects. We did this in order to establish whether or not nephron function in our subjects differs from that for residents of a temperate climate. Nineteen male and 12 female Ghanaian subjects aged between 15 and 48 years were studied. The estimated GCr was 117.3 +/- 6.6 ml/min for male subjects and 97 +/- 6.4 ml/min for female subjects. CLi was 20.3 +/- 1.6 ml/min for male and 19.1 +/- 0.4 ml/min for female subjects, respectively. The estimated absolute reabsorption rate of fluid of proximal tubules was 97.0 +/- 6.0 ml/min for males and 78.1 +/- 6.0 ml/min for females. The percentage proximal fluid reabsorption for male and female subjects was 81.2 +/- 1.4 and 79.5 +/- 1.6, respectively. The differences between male and female values (mean +/- SEM) were not statistically significant. The data suggest that the proximal tubule in residents of a tropical climate may reabsorb more fluid compared to that in residents of a temperate climate. Our values for proximal tubular reabsorption are higher than those reported for residents of a temperature climate. Our estimate of glomerular filtration, however, is similar to published data for Caucasians. The difference in proximal tubular function may reflect possible renal adaptation to a hot, humid climate. We conclude that renal function of tropical residents differs from that of residents of a temperate climate. This difference may be due to renal adaptation to the hot, tropical climate. PMID- 9334575 TI - Advances in angiogenesis research: relevance to urological oncology. AB - PURPOSE: Important advances in angiogenesis research are reviewed along with recent data implicating angiogenesis in the pathogenesis of urological malignancies. MATERIALS AND METHODS: The current understanding of angiogenesis and its importance in tumor biology is summarized. The rationale for anti angiogenic therapy is reviewed and the clinical experience with these agents is discussed. An extensive literature search of angiogenesis in urological malignancies was performed. RESULTS: Quantitative immunohistochemistry for endothelial antigens suggests that, as is the case with many other tumors, induction of angiogenesis contributes to the malignant phenotype of prostate and bladder carcinomas. Anti-angiogenic agents have demonstrated efficacy against urological tumors in experimental systems, and recent data suggest that these agents may also be useful for chemoprophylactic purposes. Putative angiogenesis inducers specific for each of the major urological malignancies have been identified. Quantitation of the expression of angiogenesis inducers and estimation of microvessel density have demonstrated prognostic value for urological malignancies. CONCLUSIONS: The available data indicate that angiogenesis has an important role in the progression and metastasis of urological malignancies. Preclinical data coupled with experience in other cancers indicate that combining anti-angiogenic therapy with conventional treatment modalities has the potential to improve dramatically our management of these malignancies. Further research will be needed to define the mechanisms controlling angiogenesis in urological malignancies and to determine if any of the angiogenic correlates will be of genuine clinical use. The rapid pace of research in this field suggests that this aspect of tumor biology will soon have an increasingly important role in the evaluation and treatment of urological cancers. PMID- 9334576 TI - Cytoreductive surgery before high dose interleukin-2 based therapy in patients with metastatic renal cell carcinoma. AB - PURPOSE: We defined the outcome of a strategy using cytoreductive surgery before high dose interleukin-2 (IL-2) therapy in patients with metastatic renal cell carcinoma. MATERIALS AND METHODS: During an 11-year period, 195 patients underwent cytoreductive surgery as preparation for high dose IL-2 based therapy. The renal primary and locoregional metastatic disease that could be safely resected was removed. RESULTS: Because of the large size 176 of 195 renal tumors (90%) were resected through transabdominal incision and in 45 patients (23%) a second additional significant procedure was performed. Five cases (2.6%) were unresectable and 2 (1%) perioperative deaths occurred. After surgery 121 of 195 patients (62%) were eligible for treatment with high dose IL-2 based protocols. Overall response rate to IL-2 based protocols was 18%. CONCLUSIONS: Cytoreductive surgery can be performed safely in patients with metastatic renal cell carcinoma. Although the impact of cytoreductive surgery on response to immunotherapy remains undefined, this combination of primary debulking and systemic IL-2 can result in durable complete tumor regression in some patients with metastatic renal cell carcinoma. PMID- 9334577 TI - Laparoscopic nephropexy: 3-year experience. AB - PURPOSE: Operative treatment of nephroptosis is rarely performed and is indicated only in select patients. Postoperatively approach related symptoms and morbidity often limit therapeutic success. We evaluated the technique and outcome of laparoscopic nephropexy in patients with symptomatic nephroptosis. For comparison the records of patients who underwent open nephropexy were reviewed. MATERIALS AND METHODS: Since 1993, 22 women and 1 man 16 to 56 years old (mean age 29) underwent laparoscopic nephropexy at our hospital. Preoperatively excretory urography and radioisotope renography were performed with the patient supine and erect, and careful pain evaluation was also done. For laparoscopic nephropexy the transperitoneal approach was used in all patients. The kidney was completely mobilized by dissection of the perirenal fat. The upper pole and convexity of the kidney were fixed to the muscle using single nonabsorbable sutures and an extracorporeal technique for tying. Between 1984 and 1994, 12 patients underwent open nephropexy at our hospital. The results of this group were reviewed and compared with those treated by laparoscopy. RESULTS: Mean operative time in the laparoscopy and control groups was 61 (range 40 to 85) and 49 minutes (range 28 to 70), respectively. In patients who underwent laparoscopic nephropexy mean analgesic use was 15 mg. morphine equivalent intravenously and 550 mg. ibuprofen orally. Postoperatively 3 minor complications (13%) were noted. Hospital stay was 3.7 days (range 2 to 9) and patients returned to work after 19 days (range 4 to 30). Six weeks after nephropexy excretory urography showed a correctly positioned kidney. At a mean 13-month followup pain intensity had improved in 21 patients (91%). According to these parameters laparoscopic nephropexy was superior to the open approach except for operative time. CONCLUSIONS: Laparoscopic nephropexy is safe and effective in the select group of patients in whom nephropexy is indicated. PMID- 9334578 TI - Paraneoplastic elevation of serum alkaline phosphatase in renal cell carcinoma: incidence and implication on prognosis. AB - PURPOSE: We investigated the incidence and prognostic significance of paraneoplastic elevation of serum alkaline phosphatase in patients with renal cell carcinoma. MATERIALS AND METHODS: Clinical data of 365 pathologically proved renal cell carcinoma cases were reviewed. Serum alkaline phosphatase level greater than 100 units per 1., but without obvious conditions that may cause phosphatase elevation, including metastasis to or disease of liver or bone and pregnancy, was regarded as paraneoplastic serum alkaline phosphatase elevation. Survival was evaluated using the Kaplan-Meier method. RESULTS: Of 365 patients 77 (21.1%) had paraneoplastic serum alkaline phosphatase elevation. The respective incidence from stage I to IV cases was 9.9% (16 of 161), 31.9% (15 of 47), 34.3% (23 of 67) and 25.6% (23 of 90). Patients with stage I disease had the lowest incidence but there were no statistically significant differences among stages II, III and IV disease. Of 77 patients with elevated serum alkaline phosphatase 48 had additional paraneoplastic manifestations. The disease specific 5-year survival rate in patients with normal serum alkaline phosphatase was significantly better than in patients with isolated phosphatase elevation, which in turn was better than in patients with multiple paraneoplastic syndromes (70.7 versus 50.5 versus 30.8%). Patients with persistent or recurrent elevation of serum alkaline phosphatase after radical nephrectomy had metastatic lesion or local recurrence. In some patients serum alkaline phosphatase returned to normal after nephrectomy but metastasis developed later without recurrent phosphatase elevation. CONCLUSIONS: Paraneoplastic serum alkaline phosphatase elevation in renal cell carcinoma patients implies an unfavorable prognosis, and additional paraneoplastic syndromes further worsen the prognosis. Recurrent or persistent serum alkaline phosphatase elevation after radical nephrectomy suggests distant metastasis or residual tumor. However, the return of serum alkaline phosphatase to normal does not guarantee cure of the disease. Identification of paraneoplastic serum alkaline phosphatase elevation is valuable in the prediction of outcome and postoperative followup of renal cell carcinoma patients. PMID- 9334579 TI - Pulmonary infarcts can mimic pulmonary metastases from renal cancer. AB - PURPOSE: Spontaneous regression of pulmonary metastases from renal cell carcinoma is a rare but well documented event. We present 2 recent cases that were radiographically consistent with pulmonary metastases from renal cell carcinoma but were pathologically shown to be pulmonary infarcts with no evidence of metastatic cells. Stable pulmonary infarcts can be misconstrued as metastatic disease in patients with renal cell carcinoma while resolving pulmonary infarcts may represent a subpopulation of patients with apparent spontaneous regression. Clinical implications of these findings are discussed. MATERIALS AND METHODS: Clinical and pathological data from 2 patients with large primary renal tumors, venous thrombi and lung masses were reviewed. Data from these cases, as well as pertinent urological and pathological literature, are presented. RESULTS: Although preoperative assessment was consistent with stage IV renal cell carcinoma, pathological examination of the lung masses in these patients showed no evidence of tumor cells. CONCLUSIONS: Pulmonary infarcts may mimic resolving or stable pulmonary metastasis in patients with renal cell carcinoma. Accurate clinical staging is crucial for the prognosis and treatment of renal cell carcinoma. Mistaking pulmonary infarcts for metastatic lesions can lead to inaccurate prognoses and inappropriate treatment. PMID- 9334580 TI - Nephrectomy before interleukin-2 therapy for patients with metastatic renal cell carcinoma. AB - PURPOSE: The management of metastatic renal cell carcinoma remains challenging and controversial. There is some evidence of improved response to interleukin-2 (IL-2) based immunotherapy in patients who undergo nephrectomy before systemic treatment. However, recent reports have suggested that surgery prior to immunotherapy may not be an efficient strategy, since many patients will not be able to receive systemic treatment after nephrectomy. We describe our criteria for determining which patients are candidates for nephrectomy before immunotherapy and present our series of patients treated with this approach. MATERIALS AND METHODS: Based on our initial experience with IL-2 based immunotherapy we developed certain inclusion criteria for treatment with initial nephrectomy followed by systemic immunotherapy, including greater than 75% debulking of tumor burden possible, no central nervous system, bone or liver metastases, adequate pulmonary and cardiac function, and Eastern Cooperative Oncology Group performance status of 0 or 1. In addition, patients in whom biopsies show other than predominantly clear cell type histology are excluded. From 1991 through 1996, 28 patients met these criteria and were treated with this approach. Patients were followed to determine the number receiving immunotherapy as well as overall response and survival rates. RESULTS: Radical nephrectomy was performed in all patients. One patient died of respiratory failure from disease progression 1 month after nephrectomy. Another patient had poor pulmonary function and, therefore, was treated with an alternative cytokine therapy. The remaining 26 patients (93%) received at least 1 course of IL-2. Median interval between nephrectomy and initiation of immunotherapy was 1.5 months (range 1 to 3). Overall response rate was 39% with 5 complete (18%) and 6 partial (21%) responses. Actuarial median survival of the entire group was 20.5 months (range 1 to 66) from the initiation of treatment. Currently 13 patients are alive, including 8 who are disease and/or progression-free. CONCLUSIONS: Using these strict criteria nephrectomy can be effectively performed before immunotherapy without compromising the likelihood that patients will receive systemic treatment. The activity of IL-2 in patients treated with this approach is encouraging and justifies its consideration in properly selected patients. PMID- 9334581 TI - Impact of 3-dimensional helical computerized tomography on selection of operative methods for ureteropelvic junction obstruction. AB - PURPOSE: We studied the feasibility of imaging the direct correlation between crossing vessels and obstructed ureteropelvic junction with helical (spiral) computerized tomography (CT) for selecting surgical repair of symptomatic ureteropelvic junction obstruction. MATERIALS AND METHODS: From July 1995 to December 1995, 4 select patients with symptomatic ureteropelvic junction obstruction underwent contrast enhanced helical CT. In addition to transaxial images, 3-dimensional reformatted images were used for evaluation. RESULTS: We identified 2 cases of ureteropelvic junction obstruction due to crossing vessels regarded as ureterovascular hydronephrosis, which is characterized by the spatial relationship between malrotated renal pelvis and anterior crossing vessels. Laparoscopic or open repair was performed in these 2 patients and operative findings were in agreement with prospective helical CT interpretation. Antegrade endopyelotomy was performed successfully for the remaining 2 patients. CONCLUSIONS: The 3-dimensional helical CT is reliable in detecting ureterovascular hydronephrosis preoperatively and in presenting better operative methods for ureteropelvic junction obstruction. PMID- 9334582 TI - Conservative surgery for ureteral tumor associated with horseshoe kidney. AB - PURPOSE: We demonstrate the successful conservative management of a ureteral tumor in a horseshoe kidney. MATERIALS AND METHODS: A patient with low grade transitional cell carcinoma of the left lower ureter had asymptomatic horseshoe kidney. Biopsy specimen revealed low grade tumor in the lower urinary tract that was associated with a congenital abnormality. The patient underwent total ureterectomy with bladder cuff excision and ileal ureteral interposition. RESULTS: With this technique renal function was preserved without resection through the isthmus of the kidney. The patient has no evidence of recurrent disease after 20-month followup by cytology, computerized tomography, excretory urogram and cystoscopy. CONCLUSIONS: Organ preserving surgery is an alternative to total nephroureterectomy in lower ureteral tumors in select patients. PMID- 9334583 TI - Long-term metabolic advantages of a gastrointestinal composite urinary reservoir. AB - PURPOSE: We investigated the long-term metabolic impact of gastrointestinal composite reservoirs. MATERIALS AND METHODS: Nine patients underwent construction of a gastroileal (7) or gastrocolonic (2) reservoir for continent urinary diversion. Four cases of metabolic acidosis were converted from a preexisting conduit and the other 5 patients had diversion for either preexisting metabolic acidosis or the short bowel syndrome. All were reconstructed using a medium sized gastric segment (8 x 4 cm.) from the greater curvature of the stomach. The anti incontinence segment was constructed from a tapered and reimplanted ileal segment. All patients underwent preoperative and postoperative measurements of serum pH, serum electrolytes, and urinalysis. Serum gastrin was measured in all patients postoperatively. Followup from surgery ranged from 47 to 61 months (mean 54.4). RESULTS: All 9 patients demonstrated electrolyte neutrality in serum on long-term followup. Postoperative serum pH (mean 7.40) was significantly different (p < 0.01) from preoperative serum pH (mean 7.36) and serum bicarbonate was also significantly different (p < 0.01) preoperatively versus postoperatively (mean 22.3 versus 25.14). Urine pH values were not significantly different throughout the study. One patient with mildly acidic urinary pH (6.0 to 6.5) had ulcerative skin changes at the stoma site. Three patients had elevated serum gastrin levels on short-term followup but all patients had normal serum gastrin levels on long-term followup. One patient, with persistent alkaline urine, had urolithiasis and symptomatic urinary tract infections. CONCLUSIONS: Our results demonstrate that a composite urinary reservoir constructed using gastric and intestinal segments achieved serum electrolyte neutrality on long-term followup. These results indicate a long-term metabolic advantage over other intestinal reservoirs associated with hyperchloremic metabolic acidosis and may be beneficial in patients compromised by either preexisting metabolic acidosis or the short bowel syndrome. PMID- 9334584 TI - Complications related to different continence mechanisms in ileocecal reservoirs. AB - PURPOSE: We compared the incidence, treatment and outcome of complications related to different continence mechanisms in a single institutional series of continent urinary diversions using an ileocecal reservoir. MATERIALS AND METHODS: From November 1990 through October 1996 in 193 consecutive cases an ileocecal pouch (Mainz I) was used as a low pressure, high capacity reservoir. A submucosally embedded in situ appendix was used in 96 patients (mean age 57.2 years, mean followup 35.6 months) and an ileal intussusception valve was used in 106 (mean age 58.4, mean followup 33.1 months). Without exception the stoma was placed in the umbilicus. RESULTS: In 172 patients (85.2%) no stoma related complication was observed. In 17 patients (17.7%) with appendix stoma 23 reinterventions were performed, for appendico-umbilical stenosis in all but 2 cases (15.6%), occurring after a mean of 20.4 months. Two complete appendix necroses required replacement by ileal nipple. Stomal stenoses could be corrected as minor outpatient procedures. In 13 of 106 patients (12.3%) with intussuscepted ileal nipple a second operation became necessary after a mean interval of 9.6 months (partial/complete necrosis of nipple in 4 cases, dislocation of nipple from ileocecal valve in 3, detachment from fascia in 4 and stomal stenosis in 2). Whereas no calculi were observed in the appendix group, stones had to be removed from 3 patients (2.8%) with ileal nipple. CONCLUSIONS: In situ appendix and intussuscepted ileal valve techniques are satisfactory in providing ileocecal reservoir continence. Besides the known advantages of the appendix as the primary reconstructive approach, the treatment of subsequent complications is simple. Therefore, whenever an appropriate appendix is encountered it should be the intestinal segment of choice in forming a continence mechanism. PMID- 9334585 TI - The results of laparoscopic adhesiolysis for intractable urinary frequency. AB - PURPOSE: We evaluated urinary frequency related to pelvic adhesions around the bladder serosa and the result of laparoscopic adhesiolysis on intractable urinary frequency caused by adhesions that did not respond to conservative treatment. MATERIALS AND METHODS: Laparoscopic adhesiolysis was performed for the diagnosis and treatment of 10 referred patients with a history of pelvic surgery in whom conservative treatment had failed. Urinary symptoms and signs, a urinary diary and cystometry were followed before and after surgery. RESULTS: In all cases there were various degrees of pelvic adhesion, mainly around the bladder. Frequency and urgency were improved in 9 of 10 patients. Urinary diary and cystometry parameters showed improvement at followup. CONCLUSIONS: Adhesions involving the bladder wall may restrict bladder distension in most of these patients. Laparoscopic adhesiolysis is beneficial in the diagnosis and treatment of this subgroup of patients. PMID- 9334586 TI - Loss of the CDKN2A/p16 locus detected in bladder irrigation specimens by fluorescence in situ hybridization. AB - PURPOSE: We measured the CDKN2A/p16 tumor suppressor gene locus in bladder irrigation specimens and correlated the measurement with the clinical status of patients with bladder cancer. MATERIALS AND METHODS: Irrigation specimens were obtained at cystoscopy from 10 normal controls, 21 patients with bladder cancer in whom no concurrent bladder tumor was seen and 23 patients with bladder tumors. Deoxyribonucleic cytometry and fluorescence in situ hybridization measurements were made. One fluorescence in situ hybridization probe was specific to the chromosome 9 centromere and the other, COSp16, targeted the CDKN2A/p16 region on chromosome 9p21. Three rates were calculated, including the hyperdiploid fraction from deoxyribonucleic acid cytometry, disomic fraction from the 9 centromere count and COSp16F, the frequency of COSp16 in association with 9 centromere. Specimens were classified as positive or negative for each of these rates using cutoff points based on previous studies and the distribution of values obtained for the normal control specimens. RESULTS: Hyperdiploid fraction values were positive (greater than 8%) in 1 normal and 1 nontumor specimen. Ten specimens from patients with tumor showed elevated hyperdiploid fraction values. In 4 nontumor and 13 tumor irrigation specimens the chromosome 9 disomic fraction values were positive (less than 80%). COSp16F was positive (less than 83%) for 18 nontumor irrigation specimens and 18 tumor irrigation specimens. One normal, and 39 of 44 nontumor and tumor irrigation specimens were positive by at least 1 test. CONCLUSIONS: COSp16 loss is measurable in irrigation specimens and it correlates with clinical status. This assay may prove useful in screening for and managing bladder cancer. PMID- 9334587 TI - Adenocarcinoma of the urachus and bladder expresses a unique colonic epithelial epitope: an immunohistochemical study. AB - PURPOSE: Primary adenocarcinoma of the bladder is a rare neoplasm whose histogenesis is poorly understood. Current data support the concept that adenocarcinoma of the bladder and urachus evolves from zones of intestinal metaplasia that become dysplastic and invasive. To address this hypothesis further we determined the immunoreactivity of benign and malignant epithelial tissue from the bladder and urachus with a monoclonal antibody that is reactive with colonic epithelium to evaluate the presence of a common reactive epitope. MATERIALS AND METHODS: The monoclonal antibody 7E12H12 (IgM isotype), developed against a colonic epithelial protein, was used in an immunoperoxidase assay to survey formalin fixed, paraffin embedded archival tissue specimens. A total of 26 specimens obtained by endoscopic biopsy or extirpative surgery, including benign and malignant bladder and urachal epithelial abnormalities, was chosen for retrospective evaluation. RESULTS: All adenocarcinoma reacted positively regardless of the histological variant, differentiation, or bladder or urachal origin. In contrast, transitional cell and squamous cell carcinomas were nonreactive. Also, the pattern of reactivity in tissues that contained benign epithelial proliferations suggested a stepwise transition with no reactivity in normal urothelium or Brunn's epithelial nests, rare staining of cystitis cystica, and uniformly positive reactivity in cystitis glandularis and frank colonic intestinal metaplasia of the bladder and urachus. CONCLUSIONS: The shared, aberrant phenotypic expression of a unique colonic epitope in benign epithelial metaplasia, and adenocarcinoma of the bladder and urachus suggests a common underlying pathway toward adenocarcinoma in cystic and urachal adenocarcinoma. The implications for diagnostic pathology are discussed. PMID- 9334588 TI - Urinary interleukin-8/creatinine level as a predictor of response to intravesical bacillus Calmette-Guerin therapy in bladder tumor patients. AB - PURPOSE: Our purpose was to determine whether urinary interleukin-8 (IL-8) levels could be used to predict a tumor-free response to intravesical bacillus Calmette Guerin (BCG) therapy in bladder cancer patients. MATERIALS AND METHODS: A total of 46 patients with superficial bladder cancer underwent an initial 6-week course of intravesical BCG therapy after transurethral resection. Voided urine samples were collected immediately before BCG instillations 1 and 6. Urine samples were centrifuged at 1,500 rpm for 8 minutes, and the supernatant was stored at -20 C. An enzyme-linked immunosorbent assay technique was used to measure urinary IL-8 levels. The Jaffe method was used to measure urinary creatinine. Results were expressed as the IL-8/creatinine ratio. Patients were followed with cystoscopy and urinary cytology every 3 months to detect bladder tumor recurrence. RESULTS: IL-8/creatinine ratios were measured in 31 patients before BCG therapy and were undetectable in 15. After 5 weeks of intravesical BCG therapy, IL-8/creatinine ratios fell in 27 patients (59%), were unchanged in 10 (22%) and rose in 9 (19%). Mean followup was 20.9 months (range 3 to 48). There was no association between the direction of change in IL-8/creatinine ratio and response to intravesical BCG therapy (p = 0.5). CONCLUSIONS: Urinary IL-8 levels obtained before intravesical BCG therapy and after instillation 5 of BCG were not helpful in predicting tumor recurrences in bladder cancer patients. PMID- 9334589 TI - Urinary incontinence and lower urinary tract symptoms: an epidemiological study of men aged 45 to 99 years. AB - PURPOSE: We assessed the prevalence of urinary incontinence and other lower urinary tract symptoms in men 45 years old or older. MATERIALS AND METHODS: A postal questionnaire was sent to a random sample (10,458) of the total population of men 45 years old or older living in Goteborg, Sweden. The response rate was 74%. RESULTS: The overall prevalence of urinary incontinence was 9.2% increasing (p < 0.001) linearly from 3.6% in men 45 years old to 28.2% in men 90 years old or older. Daily leakage was reported by 64%, and 31% believed that urinary incontinence limited their social and vocational life. However, only 46% of the men with urinary incontinence had sought medical advice. Overall prevalence of other voiding disturbances included hesitancy in 9.1% of the patients, weak stream in 30.5%, dribbling in 33.7%, sensation of incompletely emptied bladder in 26.4%, nocturia in 56.1% and history of urinary tract infection in the last 2 years in 6.3%. Prevalence of all symptoms increased with increasing age. Childhood enuresis was reported by 9.1%. Impotence was reported by 7.6% and there was a linear increase (p < 0.001) in the number of men reporting impotence up to 80 years of age. CONCLUSIONS: The prevalence of urinary incontinence and lower urogenital tract symptoms, such as hesitancy, weak stream, dribbling, sensation of incompletely emptied bladder, nocturia, urinary tract infection and impotence, increased linearly with increasing age. PMID- 9334590 TI - Results of pubovaginal sling for the treatment of intrinsic sphincteric deficiency determined by questionnaire analysis. AB - PURPOSE: We evaluated by questionnaire analysis the success rate and overall patient satisfaction after pubovaginal sling surgery. MATERIALS AND METHODS: A total of 40 women (mean age 65.7 years, range 45 to 79) underwent pubovaginal sling surgery for stress urinary incontinence due to intrinsic sphincteric deficiency. Patients completed a detailed questionnaire to assess voiding symptoms, urinary control and satisfaction. Of 40 patients 37 (92.5%) returned the questionnaire, with a mean postoperative followup of 48.2 months (range 24 to 60). RESULTS: Patients with preoperative stress urinary incontinence alone were more likely to be dry than were patients with preoperative mixed incontinence (67% versus 36%, p < 0.001). Ten patients (27%) reported stress urinary incontinence recurrence. Of the patients 23 (62.2%) reported urgency symptoms at followup, with de novo urgency occurring in 4 patients. Permanent retention was noted in 3 patients, including 2 with sacral arc denervation. Overall patients reported 86% postoperative improvement in urinary continence, and 81% would recommend the operation. CONCLUSIONS: At mean 4-year followup after pubovaginal sling surgery, this outcome study using a self administered questionnaire confirms high patient satisfaction despite significant postoperative urgency symptoms. PMID- 9334591 TI - Incisionless per vaginal bone anchor cystourethropexy for the treatment of female stress incontinence: experience with the first 50 patients. AB - PURPOSE: We evaluated the safety and efficacy of a new minimally invasive surgical procedure for the treatment of women with genuine stress urinary incontinence. MATERIALS AND METHODS: A total of 50 women (mean age 51 years) was treated for type I or II stress urinary incontinence. A miniature bone anchor and a staple like bone anchor driver were used for fixation of periurethral tissue to the public bone. RESULTS: The procedure was successfully performed in all patients without intraoperative bleeding. No significant persistent postoperative pain was noted and only 1 patient had urinary tract infection. Concomitant vaginal hysterectomy, cystocele repair or perineoplasty was performed in 33 cases. At 12-month followup 41 patients (82%) are completely continent, 7 patients (14%) reported more than 50% decrease in pad usage and 2 cases are considered surgical failures. Mean operative time was 28 minutes. CONCLUSIONS: Data suggest that our new minimally invasive procedure provides a safe, effective and easy to learn alternative for treatment of women with anatomical stress incontinence. PMID- 9334592 TI - Adverse prognostic features of collagen injection therapy for urinary incontinence following radical retropubic prostatectomy. AB - PURPOSE: We identified and characterized predictive factors associated with an unfavorable outcome of collagen injection therapy in post-radical prostatectomy incontinence. MATERIALS AND METHODS: A total of 46 patients, 49 to 85 years old (mean age 67) and incontinent after radical retropubic prostatectomy, underwent a mean of 2.8 transurethral injections of collagen (mean cumulative volume injected 31 ml.). Preoperatively, all patients underwent fluoroscopic multichannel video urodynamics including determination of Valsalva's leak point pressure. Stress urinary incontinence was subjectively graded as 1 (0 to 1 pad per day), 2 (2 to 3 pads per day) and 3 (greater than 3 pads per day). Patient age, duration and severity of pretreatment incontinence, presence of detrusor instability and anastomotic strictures, number of injections, total volume of collagen delivered and the impact of a nerve sparing procedure plus adjuvant radiation therapy were assessed and correlated with treatment outcome. RESULTS: Of the patients 11 (24%) became completely dry (9 after 3 or fewer treatments), 21 (41%) improved (17 after 3 or fewer treatments) and 14 (30%) showed no benefit (after more than 3 treatments). Of the 14 patients in whom treatment failed 6 had undergone adjuvant radiation treatment, pretreatment urinary incontinence was grade 3 in all, and concomitant detrusor instability was present in 11 (79%). All patients had received more than 3 treatments (mean total volume injected 37 ml.). CONCLUSIONS: Notwithstanding the need for multiple treatments, the prospect for cure by collagen injection of the post-radical prostatectomy incontinent patient is significantly affected by the severity of pretreatment incontinence, concomitant detrusor overactivity and exposure to radiation therapy. Age, duration of incontinence, presence of mild to moderate anastomotic strictures and a nerve sparing technique did not seem to influence treatment outcome. PMID- 9334593 TI - Urinary incontinence. PMID- 9334595 TI - Atropine role in the pharmacological erection test: study of 228 patients. AB - PURPOSE: We determined the efficacy of atropine sulfate combined with papaverine hydrochloride, prostaglandin E1 and phentolamine mesylate in the pharmacological erection test. MATERIALS AND METHODS: A total of 230 consecutive patients with erectile dysfunction was randomized for pharmacological erection test and received an intracorporeal combination of 50 mg. papaverine hydrochloride, 10 micrograms. prostaglandin E1, 0.2 mg. phentolamine mesylate and 0.075 mg. of atropine sulfate (group 1), or the same combination without atropine sulfate (group 2). Erectile response was evaluated subjectively and by intracorporeal pressure measurement. RESULTS: In group 1, 40 patients (35.1%) showed only tumescence, and 22 (19.3%) had poor erection. In group 2, 45 patients (39.5%) had tumescence and 17 (14.9%) poor erection. In both groups 52 patients (45.6%) had rigid erection. There was no statistically significant difference regarding erectile response and intracorporeal pressure. CONCLUSIONS: The addition of atropine sulfate did not improve results of the pharmacological erection test when combined with 50 mg. papverine hydrochloride, 10 micrograms, prostaglandin E1, and 0.2 mg. phentolamine mesylate. PMID- 9334594 TI - Intracavernosal forskolin: role in management of vasculogenic impotence resistant to standard 3-agent pharmacotherapy. AB - PURPOSE: We investigated forskolin, a direct adenylate cyclase activator, as an intracavernosal vasoactive agent in management of vasculogenic impotence. MATERIALS AND METHODS: Concentration responses for forskolin and prostaglandin E1 induced relaxation of phenylephrine precontracted strips of human corpus cavernosum smooth muscle were constructed in vitro. Cyclic adenosine monophosphate (cAMP) synthesis was determined with papaverine, phentolamine, prostaglandin E1 and forskolin in human corpus cavernosum smooth muscle cell cultures. Dose-dependent hemodynamic responses to intracavernosal forskolin (5 to 20 micrograms) were evaluated in a New Zealand White rabbit model. Safety and efficacy outcome data were obtained in vasculogenically impotent patients who signed informed consent and met strict inclusion and exclusion criteria that included having had standard self-injection therapies fail. RESULTS: In vitro forskolin and prostaglandin E1 alone caused concentration dependent relaxation with an EC50 of approximately 200 nm. and 16 nm., respectively. When the 2 agents were combined, the concentration response curve for relaxation shifted to the left. cAMP production was highest in cells treated with prostaglandin E1 and forskolin and was unaffected by papaverine or phentolamine. In 3 animals, equilibrium intracavernosal pressure and duration of erection had a dose dependent increase. Clinical investigation in 31 patients showed no adverse events with a mean of 14 +/- 4, range 11 to 18 months of followup. Overall 61% reported improvement in rigidity and/or erection duration using intracavernosal forskolin (98 micrograms./ml.), papaverine (29 mg./ml.), phentolamine (0.98 mg./ml.) and prostaglandin E1 (9.8 micrograms./ml.). CONCLUSIONS: Forskolin is a United States Food and Drug Administration nonapproved vasoactive agent that acts in synergism with prostaglandin E1 to induce smooth muscle relaxation. In combination with other vasoactive agents, forskolin has demonstrated preliminary safety and efficacy in patients with vasculogenic impotence resistant to standard 3-agent pharmacotherapy. PMID- 9334596 TI - Endocrine screening in 1,022 men with erectile dysfunction: clinical significance and cost-effective strategy. AB - PURPOSE: We reviewed the results of serum testosterone and prolactin determination in 1,022 patients referred because of erectile dysfunction and compared the data with history, results of physical examination, other etiological investigations and effects of endocrine therapy to refine the rules of cost-effective endocrine screening and to pinpoint actual responsibility for hormonal abnormalities. MATERIALS AND METHODS: Testosterone and prolactin were determined by radioimmunoassay. Every patient was screened for testosterone and 451 were screened for prolactin on the basis of low sexual desire, gynecomastia or testosterone less than 4 ng./ml. Determination was repeated in case of abnormal first results. Prolactin results were compared with those of a previous personal cohort of 1,340 patients with erectile dysfunction and systematic prolactin determination. Main clinical criteria tested regarding efficiency in hormone determination were low sexual desire, small testes and gynecomastia. Endocrine therapy consisted of testosterone heptylate or human chorionic gonadotropin for hypogonadism and bromocriptine for hyperprolactinemia. RESULTS: Testosterone was less than 3 ng./ml. in 107 patients but normal in 40% at repeat determination. The prevalence of repeatedly low testosterone increased with age (4% before age 50 years and 9% 50 years or older). Two pituitary tumors were discovered after testosterone determination. Most of the other low testosterone levels seemed to result from nonorganic hypothalamic dysfunction because of normal serum luteinizing hormone and prolactin and to have only a small role in erectile dysfunction (definite improvement in only 16 of 44 [36%] after androgen therapy, normal morning or nocturnal erections in 30% and definite vasculogenic contributions in 42%). Determining testosterone only in cases of low sexual desire or abnormal physical examination would have missed 40% of the cases with low testosterone, including 37% of those subsequently improved by androgen therapy. Prolactin exceeded 20 ng./ml. in 5 men and was normal in 2 at repeat determination. Only 1 prolactinoma was discovered. These data are lower than those we found during the last 2 decades (overall prolactin greater than 20 ng./ml. in 1.86% of 1,821 patients, prolactinomas in 7, 0.38%). Bromocriptine was definitely effective in cases with prolactin greater than 35 ng./ml. (8 of 12 compared to only 9 of 22 cases with prolactin between 20 and 35 ng./ml.). Testosterone was low in less than 50% of cases with prolactin greater than 35 ng./ml. CONCLUSIONS: Low prevalences and effects of low testosterone and high prolactin in erectile dysfunction cannot justify their routine determination. However, cost-effective screening strategies recommended so far missed 40 to 50% of cases improved with endocrine therapy and the pituitary tumors. We now advocate that before age 50 years testosterone be determined only in cases of low sexual desire and abnormal physical examination but that it be measured in all men older than 50 years. Prolactin should be determined only in cases of low sexual desire, gynecomastia and/or testosterone less than 4 ng./ml. PMID- 9334597 TI - Erectile dysfunction--the paradigm changes but the challenges remain. PMID- 9334598 TI - Endothelin-1 in diabetic and nondiabetic men with erectile dysfunction. AB - PURPOSE: We evaluated plasma concentration of endothelin-1 in diabetic and nondiabetic men complaining of erectile dysfunction, and the variation of endothelin-1 in cavernous body blood during intracavernous injection of prostaglandin E1. MATERIALS AND METHODS: We evaluated plasma concentrations of endothelin-1 in venous blood of 20 men with erectile dysfunction, 10 with and 10 without diabetes. Plasma concentration of endothelin-1 was also evaluated in the cavernous body blood of the 20 men with erectile dysfunction, during erection induced by intracavernous injection of 10 micrograms prostaglandin E1. A severe vasculogenic component of erectile dysfunction was excluded in all patients. RESULTS: Basal plasma concentration of endothelin-1 in the cubital vein was increased in nondiabetic (1.13 +/- 0.4 pg./ml.) and in diabetic (1.80 +/- 0.2 pg./ml.) patients with erectile dysfunction, compared to control men (0.64 +/- 0.1 pg./ml.) (p < 0.0005 and p < 0.0001, respectively), and in diabetic compared with nondiabetic patients (p < 0.002). No difference and close correlation were observed in the concentration of endothelin-1 in the cavernous body blood evaluated 5 minutes and 30 minutes after injection of prostaglandin E1 (r = 0.89, p < 0.0001, y = 0.98 x + -0.066). The concentration of endothelin-1 in the cavernous body blood evaluated 30 minutes after injection of prostaglandin E1 did not show any difference compared to peripheral venous concentration of the peptide in the 2 patient groups. Concentrations of endothelin-1 in the peripheral vein and the cavernous body blood were not different in patients with a full erection compared with incomplete penis erection after injection of prostaglandin E1 in the cavernous body. PMID- 9334599 TI - The effect of parenteral testosterone replacement on prostate specific antigen in hypogonadal men with erectile dysfunction. AB - PURPOSE: Parenteral testosterone supplementation is a common treatment for erectile dysfunction in hypogonadal men. Despite its frequent use, the effect of testosterone on prostate specific antigen (PSA) in these patients has not been documented previously. In this study we determined the effect of parenteral testosterone replacement on PSA and PSA velocity in a group of men being treated for erectile dysfunction. MATERIALS AND METHODS: A retrospective analysis of 48 patients (mean age 65.9) was performed and 2 study groups were identified. Group 1 consisted of 27 patients with a serum PSA level before and after initiating testosterone replacement therapy, and group 2 consisted of 27 men with a minimum of 3 PSA measurements (intervals of 6 months or greater) while on testosterone replacement. Each man had erectile dysfunction, a normal digital rectal examination and a low or low-normal total serum testosterone level before initiating therapy. Testosterone replacement was discontinued if no subjective improvement in erectile function was obtained, or if prostate adenocarcinoma was suggested by digital rectal examination or PSA. RESULTS: The mean increase in PSA after initiating testosterone replacement was 0.29 ng./ml. representing a mean change of 37% from baseline (mean interval 12.8 months). The mean PSA velocity was 0.05 ng./ml. per year. Pretreatment testosterone level, age and testosterone dose did not independently alter the PSA during testosterone replacement. Eleven men required prostate biopsies during treatment. Biopsies were indicated for abnormal digital rectal examination in 10 men and an elevated PSA in 1. All biopsies were benign. CONCLUSIONS: Parenteral testosterone replacement in hypogonadal men with normal pretreatment digital rectal examination and serum PSA levels does not alter PSA or PSA velocity beyond established nontreatment norms. Thus, any significant increase in PSA or PSA velocity should not be attributed to testosterone replacement therapy and should be evaluated. PMID- 9334600 TI - Biochemical alterations of the tunica albuginea in impotence. AB - PURPOSE: The objectives of this study were to quantify the immunohistochemical stainings of collagen types I, III and IV, and investigate the value of glycohistochemical staining with 3 lectin types specific to a particular glycan structure, Arachis hypogaea, Triticum vulgare and concanavalin A, as a method of defining possible changes in the collagen structure of the tunica albuginea in potent and impotent patients. MATERIALS AND METHODS: The study involved 4 normal men, 4 with pure venous leakage and 4 with pure arterial disease. Collagen types I, III and IV, and lectins Arachis hypogaea, Triticum vulgare and concanavalin A were studied using a cell image processor. The labeling index relates to the percentage of staining and mean optical density relates to the staining intensity. RESULTS: Mean labeling index values for the 3 types of collagen and lectins were similar (p > 0.05). Mean optical density value relating to collagen type I was significantly higher in the arteriogenic group than in the other groups (p < 0.05), while mean optical density value of collagen type IV was significantly higher in the venogenic group than in the 2 other groups (p < 0.05). Mean optical density values relating to the 3 lectin types were similar in the 3 clinical groups (p > 0.05). CONCLUSIONS: An alteration in the distribution and structure of the various collagen types and lectins in the tunica albuginea of impotent patients has been shown that may interfere with normal function and lead to impotence. PMID- 9334601 TI - Laparoscopically assisted penile revascularization for vasculogenic impotence: 2 additional cases. AB - PURPOSE: Microsurgical revascularization of the penis in vasculogenic impotence is an accepted surgical procedure in young men with a history of blunt pelvic or perineal trauma. Most penile revascularization techniques use the inferior epigastric artery in direct artery-to-artery revascularization or dorsal vein arterialization procedures. To obviate the wide pararectal incision laparoscopic mobilization of the inferior epigastric vessels has been recently proposed. We present 2 cases of successful laparoscopically assisted penile revascularization. MATERIALS AND METHODS: With the patient under general anesthesia the first trocar was inserted in the umbilical region and pneumoperitoneum was induced. Two other trocars were positioned laterally. As soon as the inferior epigastric vessels were accessed, dissection was initiated below the level of the arcuate line. The vessels were dissected cephalad en bloc to a point of bifurcation of the inferior epigastric artery above the umbilical level. The inferior epigastric pedicle was ligated with clips and transected at the cephalad edge of the dissection. It was then mobilized and tunneled through an infrapubic incision at the base of the penis for subsequent microvascular anastomosis with the penile vessels. RESULTS: The anastomosis was patent and hemostasis was satisfactory. Operative time in the 2 cases was 4.3 and 5.2 hours, respectively. At 3 months both patients reported complete erections. CONCLUSIONS: Our experience confirms the extremely practical use of laparoscopy which, due to its magnification power, makes it possible to perform fast, accurate excision of the epigastric bundle. Moreover, a wide pararectal incision, which is a frequent cause of postoperative complications, is avoided. PMID- 9334602 TI - Outcome analysis of penile implant surgery after external beam radiation for prostate cancer. AB - PURPOSE: We evaluated the success and possible complication rates of penile implant surgery in patients who underwent external beam radiation therapy for prostate cancer. MATERIALS AND METHODS: We reviewed the charts of 43 patients who underwent penile implant surgery after radiation therapy for prostate cancer. The type, dose and volume of radiation were assessed. The types of surgical approach and prosthesis as well as complications were recorded. A total of 34 patients was alive and traceable, and 9 were untraceable (7 dead and 2 missing). The 34 traceable patients were interviewed personally or by telephone to evaluate the function of and satisfaction with the penile implant. Followup of the 9 untraceable patients was assessed through a chart review. RESULTS: A total of 35 patients (81%) received definitive radiation therapy to the prostate and seminal vesicles, and 8 (19%) underwent radical retropubic prostatectomy followed by radiation therapy to the prostatic bed. Mean age at implant surgery was 67 years (range 36 to 83). In the 43 men 46 procedures were done and mean followup was 40 months. None of the patients in this series had infection or erosion. Of the men 24 (71%) use the prosthesis at least once weekly or more for sexual intercourse, 6 (17%) use it twice monthly, 4 (12%) are not sexually active despite a functioning implant, 2 are not sexually active because of a lack of sexual partners, and 2 are not satisfied with the implant and would not recommend this device. Discomfort from the penile implant was reported by 2 patients, although they currently use the implant for intercourse at least twice weekly. CONCLUSIONS: Penile prosthesis surgery can be safely and effectively performed after radiation therapy with minimal intraoperative and postoperative complications, and an excellent patient satisfaction rate. PMID- 9334603 TI - Apoptotic frequency is increased in spermatogenic maturation arrest and hypospermatogenic states. AB - PURPOSE: Increased testicular apoptosis has been observed in maturation arrest and hyposper-matogenesis states in rodent models, but this process has not yet been characterized in humans. We hypothesized that increased cell death present with accelerated apoptosis is significant in pathophysiology of many male infertility states associated with abnormal spermatogenesis. We examined frequency of apoptotic bodies in human testis biopsy specimens from infertile men using morphometric analysis of hematoxylin and eosin stained paraffin sections. MATERIALS AND METHODS: Testis biopsy specimens were obtained for routine clinical purposes from azoospermic and severely oligozoospermic men and were stained with hematoxylin and eosin. Apoptotic bodies were identified using established morphometric criteria. Apoptotic indexes, defined as apoptotic bodies per total number of cells and per Sertoli cells, were calculated after counting all intratubular spermatogenic cells and Sertoli cells in 20 tubules. RESULTS: A total of 51 biopsies was performed in 50 men. Significantly increased apoptotic body per total cell and apoptotic body per Sertoli cell ratios were observed in maturation arrest and hypospermatogenesis states in comparison to Sertoli cell only and normal spermatogenesis (p < 0.05, Mann-Whitney test). CONCLUSIONS: Increased apoptosis in maturation arrest and hypospermatogenesis states compared to normal but obstructed spermatogenesis and Sertoli cell only were observed, indicating a prominent role for this form of programmed cell death in human male infertility. PMID- 9334604 TI - Congenital absence of the vas deferens: incomplete penetrance of cystic fibrosis gene mutations. AB - PURPOSE: We evaluated the penetrance of cystic fibrosis gene mutations for the clinical phenotype of congenital bilateral absence of the vas deferens. MATERIALS AND METHODS: We retrospectively reviewed the fertility status of 244 brothers of 105 men with congenital bilateral absence of the vas deferens. Testing for the most common cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations and for haplotype analysis of the intron 8/polythymidine segment was recommended for all men with congenital bilateral absence of the vas deferens. RESULTS: Of 244 brothers of men with congenital bilateral absence of the vas deferens 131 were eligible for assessment of fertility. Of the 131 evaluable brothers only 7 (5%) were found to have congenital bilateral absence of the vas deferens. This prevalence is 5 times lower than that predicted for congenital bilateral absence of the vas deferens (25%) based on the autosomal recessive inheritance pattern seen in classical cystic fibrosis. For couples in which the man has congenital bilateral absence of the vas deferens and the female partner tests negative for standard CFTR gene mutations including 5T analysis, the maximum risk of having a child with congenital bilateral absence of the vas deferens is less than 1.0%. CONCLUSIONS: Our data are consistent with incomplete penetrance for the congenital bilateral absence of the vas deferens phenotype after inheritance of cystic fibrosis gene mutations, even after adjusting for environmental factors and wolffian anomalies. Incomplete penetrance may account for a low prevalence of congenital bilateral absence of the vas deferens in the population and may lower the risk of having a child with congenital bilateral absence of the vas deferens for couples undergoing sperm retrieval and assisted reproductive techniques. PMID- 9334605 TI - Effects of subinguinal varicocele ligation on sperm concentration, motility and Kruger morphology. AB - PURPOSE: We examined the effects of varicocelectomy on semen parameters in 30 subfertile men, with emphasis on potential changes in sperm count, motility and morphology as measured by Kruger's strict morphologic criteria. MATERIALS AND METHODS: A total of 30 patients underwent subinguinal varicocelectomy (25 bilateral and 5 unilateral). Preoperative and postoperative sperm density, motility and morphology were analyzed. Preoperative follicle-stimulating hormone, luteinizing hormone and testosterone levels were measured and compared to those of fertile volunteers enrolled in our sperm donation program. Pregnancy rates after varicocelectomy were also examined: The Wilcoxon signed rank test was used to measure levels of statistical significance in all analyses. RESULTS: We found that sperm density and motility improved significantly (p < 0.05) without concomitant changes in strict morphology (p > 0.05) only in men with clinical bilateral varicoceles. No differences were observed in values among testosterone, follicle-stimulating hormone and luteinizing hormone levels of the fertile control group and preoperative varicocele patients. Of 30 patients 12 (40%) had successful, full-term pregnancies, including 6 via natural cycle intercourse, 5 (43%) by in vitro fertilization embryo transfer and 1 by intracytoplasmic sperm injection. CONCLUSIONS: Although sperm morphology as measured by strict morphologic criteria does not improve after varicocelectomy, there were highly significant changes in motility and concentration. Hormonal differences are not likely to have a role in or be reflective of pathophysiology of varicocele induced male infertility. The recent observation that sperm motility may be an independent or additive predictive factor for fertilization and pregnancy supports the need for continued varicocele repair independent of the lack of varicocelectomy effect on Kruger morphology. PMID- 9334607 TI - New insights into the etiology and treatment of male infertility. PMID- 9334606 TI - Response of routine semen analysis and critical assessment of sperm morphology by Kruger classification to therapeutic varicocelectomy. AB - PURPOSE: We studied the effect of varicocelectomy on Kruger morphology and semen parameters. MATERIALS AND METHODS: A total of 33 subfertile men diagnosed with varicoceles was evaluated 3 months before, and 3 to 4 and 6 to 8 months after varicocelectomy. Evaluation involved routine semen analysis and sperm morphology using Kruger classification. RESULTS: Significant improvement in sperm concentration and count was found after varicocelectomy (sperm count preoperatively 117.1 +/- 29, 3 to 4 months postoperatively 162.5 +/- 41 and 6 to 8 months postoperatively 139.8 +/- 25 million sperm, p = 0.0095). Using Kruger classification, evaluation of sperm morphology revealed overall significant increase in percentage of normal A forms at 3 to 4 and 6 to 8 months after surgery (from 9.8 +/- 5.8% A forms, 13.6 +/- 7.7% A forms, and 14.5 +/- 7.5% A forms, respectively, p = 0.0002, normal greater than 14%). Twelve of the 26 patients (46%) with abnormal sperm morphology preoperatively and greater than 4% A forms reached normal levels 3 months postoperatively. Six months after surgery only 6 patients maintained normal values and 3 of the initial 14 nonresponders became normal (9 of 26, 36%). Three patients with severe teratozoospermia (less than 4% A forms) showed improvement in sperm morphology. Four patients with normal sperm morphology preoperatively were not affected by varicocelectomy. CONCLUSIONS: Surgical correction of varicocele was associated with significant improvement in sperm morphology evaluated using Kruger classification. Concentration and count improved after varicocelectomy. Changes were observed as early as 3 months after surgery. PMID- 9334608 TI - Treatment of condylomata acuminata with oral isotretinoin. AB - PURPOSE: The aim of our study was to evaluate the efficacy and safety of oral isotretinoin in the treatment of condylomata acuminata. MATERIALS AND METHODS: A total of 56 male patients with a history of condylomata acuminata refractory to at least 1 standard therapeutic regimen was treated orally with isotretinoin (1 mg./kg. daily) during a 3-month period. RESULTS: At the end of treatment 21 of the 53 evaluated patients (39.6%) had complete response, 7 (13.2%) had partial response and 25 (47.1%) had no response. A statistically significant inverse relationship was found between age and area of treated lesions and response to medication. Two complete responders (9.5%) revealed recurrence during the 1-year followup. CONCLUSIONS: Oral isotretinoin may be regarded as an effective, fairly well tolerated and noninvasive alternative form of therapy for immature and small condylomata acuminata. PMID- 9334609 TI - Vena caval resection for bulky metastatic germ cell tumors: an 18-year experience. AB - PURPOSE: The operative management and followup of vena caval resection for bulky metastatic germ cell tumors have been previously described in 3 series. In 1989 Ahlering and Skinner described their experience with 12 patients. We now update this experience with the most recent followup on 19 patients. MATERIALS AND METHODS: From April 1978 to May 1995, 19 men underwent retroperitoneal lymph node dissection for stage B3 (N3) or C (N3, M+) germ cell tumor after induction chemotherapy. In all cases the inferior vena cava was resected because of extensive thrombosis or direct involvement of the vessel wall by a tumor. The inferior vena cava was resected from just below the renal veins to beyond the level of disease involvement. Complete resection of retroperitoneal disease was accomplished in all patients. Morbidity and mortality were examined. RESULTS: The mean hospital stay was 10 days (range 7 to 13) for uncomplicated recoveries (9 patients) versus 19 days (range 6 to 32) for complicated recoveries (10 patients). Followup ranged from 1 month to 16 years. Complications included prolonged ileus, small bowel obstruction, fascial dehiscence and pneumonia with pleural effusion. Chronic edema persisted in 3 of 11 patients with followup of greater than 6 months. Of the 6 patients who died of disease recurrence 4 did not have normalization of tumor markers before surgery, and all 4 had persistence of cancer in the resected specimen. Seven patients are without disease at followup of 24 months to 16 years. All survivors had normalized tumor markers before surgery. Only 1 patient (5%) had retroperitoneal recurrence. CONCLUSIONS: En bloc vena caval resection for tumor involvement or extensive thrombosis can be associated with short and long-term morbidity, is feasible, and may contribute to a prolonged tumor-free interval and a chance for cure. PMID- 9334610 TI - Immunohistochemical and ultrastructural study of rhabdosphincter component of the prostatic capsule. AB - PURPOSE: There has been no complete agreement on functional anatomy of muscular components of the urethral sphincteric mechanism, particularly in the male patient. The prostatic capsule was studied to define its histological structure and to determine whether its rhabdosphincter component (prostatocapsular rhabdosphincter) consists only of slow twitch or slow and fast twitch striated myofibers. MATERIALS AND METHODS: We studied 11 whole prostates, including 1 obtained at autopsy and 10 by radical prostatectomy. Samples of prostatic capsule from 4 operative specimens were studied by electron microscopy. Whole mount paraffin sections from transverse slices of the remaining 7 prostates were double labeled with avidin biotin conjugate immunostaining using the primary monoclonal antibodies anti-alpha smooth muscle actin plus anti-alpha sarcomeric actin (all striated myofibers) or antiskeletal myosin fast (fast myofibers only). Tissue components of the prostatic capsule, including smooth muscle and slow versus fast twitch striated myofibers, were quantified by computerized image analysis. RESULTS: The prostatic capsule consisted of collagen, smooth muscle and striated myofibers. It varied in thickness and proportion of the 3 components among specimens, and in each in relation to transverse circumferential aspect and craniocaudal (horizontal) level of the prostate. Collagen and smooth muscle were equally important components. Striated muscle elements within the capsule consisted of fast twitch and dominant slow twitch myofibers, and were much more abundant in the caudal (distal, lower) than the cranial (proximal, upper) half of the capsule, where they were deficient ventrally (anteriorly) and dorsally (posteriorly). The prostatocapsular rhabdosphincter thus had a butterfly-like appearance, with a thick posteriorly open ring at the apex and 2 thinner, divergent leaflets tapering toward the base at the bladder neck. The fast myofiber population decreased progressively from apex to base of prostate. CONCLUSIONS: Proof is provided for mixed slow and fast twitch myofiber structure of the prostatocapsular component of human male rhabdosphincter. Sustained (tonic) contraction of slow myofibers probably reinforces the role of urethral smooth muscle in maintaining continence during bladder filling. Swift contraction of fast myofibers that abound caudally in the capsule probably supplements urethral closure by the bulkier membranous urethral part of the rhabdosphincter in preventing leakage of urine under stress when voiding is imminent or willfully withheld. PMID- 9334611 TI - Urodynamic assessment of patients with acute urinary retention: is treatment failure after prostatectomy predictable? AB - PURPOSE: Some patients with acute urinary retention due to benign prostatic hyperplasia do not have successful outcome after prostatectomy and require either a chronic indwelling urethral catheter or clean intermittent catheterization. Urodynamic and clinical parameters were examined preoperatively in 81 men 56 to 93 years old (mean age 72 years) in search of an outcome predictor after prostatectomy. MATERIALS AND METHODS: International Prostate Symptom Score, prostate volume, retention episodes, retention volume and urodynamic parameters from a multichannel pressure-flow study were analyzed preoperatively and postoperatively. All patients underwent transurethral prostatectomy and were reexamined 2, 4, 12 and 24 weeks after surgery. A multichannel pressure-flow study was performed preoperatively and 12 weeks postoperatively. RESULTS: At 24 weeks postoperatively 11 patients (13%) were unable to void and therefore classified as treatment failures while the remaining patients voided spontaneously and were classified as treatment successes. There were statistically significant differences (p < 0.005) between treatment failure and treatment success regarding age (83.5 +/- 7 versus 70.1 +/- 8 years), preoperative volume of retention (1,780 versus 1,080 ml.), and maximal detrusor pressure (24.4 versus 73.5 cm. water), but not to International Prostate Symptom Score, episodes of retention and prostate volume. The ability to void during preoperative pressure flow study and the presence of detrusor instability predicted good outcome. In treatment success patients postoperative urodynamic data showed significant decrease in detrusor pressure at maximum flow rate (from 80.8 +/- 33 to 34.6 +/- 10 cm. water). Those with treatment failure had an increase in maximal detrusor pressure (from 26 +/- 12 to 42.6 +/- 13 cm. water), suggesting detrusor recovery. CONCLUSIONS: Patients with acute urinary retention, age 80 years or older, with retention volume greater than 1,500 ml., no evidence of instability and maximal detrusor pressure less than 28 cm. water are at high risk of treatment failure. However, despite treatment failure the detrusor may recover in patients younger than 80. Therefore, prostatectomy should still be performed in this group (less than 80 years old) even if preoperative urodynamics suggest an unfavorable outcome. PMID- 9334612 TI - Transurethral needle ablation of the prostate: a urodynamic based study with 2 year followup. AB - PURPOSE: We evaluated the efficacy of transurethral needle ablation of the prostate for the treatment of lower urinary tract symptoms related to benign prostatic hyperplasia (BPH). This study was urodynamic based with 2-year followup to determine whether transurethral needle ablation of the prostate could reduce bladder outlet obstruction and, if so, whether the effect was durable. MATERIALS AND METHODS: A total of 47 patients with symptomatic BPH underwent transurethral needle ablation of the prostate under local anesthesia and intravenous sedation. All patients were evaluated subjectively using the American Urological Association symptom index and the quality of life score. Patients were evaluated objectively with uroflowmetry, post-void residual volume and pressure-flow studies. All patients underwent subjective and objective evaluation before treatment. Followup was conducted at 1, 3, 6, 12 and 24 months after treatment. Short and long-term complications were assessed. RESULTS: At 6-month followup there was 71% improvement in mean cases (22.4 to 6.6, 42 patients symptom index, p < 0.05), and 66% improvement in mean quality of life score (4.6 to 1.56, 42 patients, p < 0.05). Maximum flow rate, post-void residual volume and detrusor pressure at maximum flow rate also showed statistically significant improvements throughout the study. At 12-month followup there was a 55% increase in maximum flow rate (6.6 to 10.23 ml. per second, 29 patients, p < 0.05). A 37% reduction in mean detrusor pressure at maximum flow rate (92.4 cm. to 58 cm. water, 31 patients, p < 0.05) was recorded at 24-month followup, thus indicating that transurethral needle ablation of the prostate can lower bladder pressure significantly. Post-void residual volume decreased from a pretreatment mean of 76.1 ml. to a mean of 36.9 ml. (31 patients, p < 0.05) at 24 months. Short-term complications (3 months) included transient posttreatment urinary retention in 8 patients (17%), duration 1 to 9 days, mild to moderate transient frequency dysuria all patients which resolved in more than 90% by 5 weeks and epididymitis in 1. A patient questionnaire was used to evaluate changes in sexual function and there were no reports of disturbances in erectile function or retrograde ejaculation. There were no long-term complications. However, 6 patients (12.7%) had persistent bothersome symptoms during the followup period and underwent transurethral prostate resection. Further analysis of this subset of patients with respect to pretreatment evaluation and transurethral needle ablation procedure did not reveal significant differences between them and patients with successful outcomes. CONCLUSIONS: Transurethral prostate resection is a safe and effective technique for treating lower urinary tract symptoms related to benign prostatic hyperplasia. The technique can be performed in the office as an outpatient, or as a same day surgical procedure, using topical anesthesia with intravenous sedation, if necessary. In the majority of patients subjective and objective improvements were sustained for the duration of this study, which included 2-year followup with pressure-flow studies. PMID- 9334613 TI - Quality of life assessment in patients treated with lower energy thermotherapy (Prostasoft 2.0): results of a randomized transurethral microwave thermotherapy versus sham study. AB - PURPOSE: We evaluated the impact of lower energy transurethral microwave thermotherapy on quality of life and quality of sexual function in patients with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 50 patients with BPH were randomized to receive either lower energy transurethral microwave thermotherapy treatment (Prostasoft 2.0) or placebo treatment and followed for 26 weeks after treatment. All patients completed a Madsen symptom score and quality of life questionnaire to assess acceptability, daily activities, psychological well-being, social activities and improvement in quality of life. A sexual function questionnaire was used to assess changes in sexual function after microwave thermotherapy. RESULTS: A significant difference in voiding parameters and symptom score was found between the transurethral microwave thermotherapy and sham groups. Maximum uroflow changed from 9.6 ml. per second at baseline to 13.9 ml. per second and from 9.9 ml. per second at baseline to 9.6 ml. per second at 26 weeks for transurethral microwave thermotherapy and sham groups, respectively. Madsen score improved from 13.2 to 5.3 for the transurethral microwave thermotherapy group and from 11.9 to 9.1 for the sham group. For quality of life measures, a statistically significant difference in favor of the transurethral microwave thermotherapy group was found only for the acceptability item. At baseline and after 26 weeks no statistically significant difference was observed between the 2 groups for Quality of Life measures documenting sexual function. However, almost 20% of patients treated by either transurethral microwave thermotherapy or sham claimed at 26 weeks after treatment that treatment had influenced sexual function. CONCLUSIONS: Although significant changes in objective and subjective parameters were found in patients after lower energy microwave thermotherapy, the change in quality of life was minimal. In addition to the minimal invasiveness of transurethral microwave thermotherapy, preservation of sexual function is appealing. PMID- 9334614 TI - Secular trend and age-period-cohort analysis of prostate cancer mortality in Taiwan. AB - PURPOSE: We evaluated the secular mortality trend of prostate cancer in Taiwan from 1964 through 1994. MATERIALS AND METHODS: Analyses were based on vital statistics. Relative risks associated with each of the age, period and cohort effects on secular mortality were estimated from a log-linear Poisson model. RESULTS: Age adjusted mortality rates increased more than 2-fold during the last 30 years in Taiwan. Age-period-cohort analysis showed that the age effect was the strongest. CONCLUSIONS: Factors related to aging are the main reason for the increase in prostate cancer mortality in Taiwan during the least 3 decades. PMID- 9334615 TI - Trends in diagnosis of stage T1a-b prostate cancer. AB - PURPOSE: Stage T1a-b prostate cancer comprised about 44% of newly diagnosed local prostate cancer cases in the United States before the advent of medical and minimally invasive treatments for symptomatic benign prostatic hyperplasia (BPH) and before the widespread use of prostate specific antigen (PSA) testing in men with BPH. Information about the impact of these advances on detection of T1a-b cancer is not available. MATERIALS AND METHODS: Prevalence of T1a-b prostate cancer was determined in 1,554 consecutive men who underwent surgical prostatectomy for suspected BPH at a Veterans Affairs Medical Center from 1985 through 1996. Since 1991 a PSA blood test was obtained routinely before surgery and patients with PSA greater than 4.0 ng./ml. usually underwent ultrasound guided prostate biopsy. RESULTS: The number of T1a-b cancer cases was relatively stable during 1985 to 1990 but declined from 36 in 1990 to 9 in 1996. There were no temporal trends in proportion of prostatectomy patients with T1a-b cancer and the decline in cancer detection paralleled less frequent use of surgical prostatectomy for treatment of BPH. The proportion of prostatectomy patients with T1a-b cancer was similar in 1985 to 1990 and in 1991 to 1996 but the percentage of Gleason 7 to 10 cancers declined from 26 in 1985 to 1990 to 10 in 1991 to 1996 (p < 0.0001). PSA and PSA density of evaluable patients with cancer were significantly greater than in evaluable patients with BPH. Of 105 patients with PSA greater than 4.0 ng./ml. who underwent preoperative prostate biopsy 16 (15%) had T1a-b cancer. CONCLUSIONS: The less frequent use of surgical prostatectomy at our institution has produced marked decline in detection of T1a-b cancer. If representative of national trends this experience suggests that many men with obstructive voiding symptoms and T1a-b cancer will remain undiagnosed and that periodic monitoring to identify unsuspected cancer is important in men who are treated with medical or minimally invasive therapies for BPH. Decline in detection of T1a-b cancer may also confound the accuracy of projected incidence rates of local prostate cancer in the United States. PMID- 9334617 TI - Prostate cancer 1997--practicing what we preach. PMID- 9334616 TI - Current trends in prostate cancer diagnosis and staging among United States urologists. AB - PURPOSE: We analyzed current practice patterns and determined whether urologists are diagnosing and staging prostate cancer in accordance with one another and with available literature. MATERIALS AND METHODS: An anonymous questionnaire was mailed to 1,500 randomly selected practicing American Urological Association members throughout the United States, categorized according to practice setting and decade of residency training completion. RESULTS: There were 624 respondents (41.6%). Annual routine prostate cancer detection is being aimed toward the right of the age spectrum. More than half of respondents use age specific prostate specific antigen (PSA), while fewer than half use PSA density in determining need for biopsy. The vast majority will perform radical prostatectomy on patients whose age suggests that they will not benefit from surgery. High PSA values and Gleason scores often are disregarded as independent precluding factors when deciding to perform radical prostatectomy. Computerized tomography and radionuclide bone imaging are used routinely far in excess of what the literature suggests is appropriate. Regardless of preoperative staging results, most urologists still perform lymphadenectomy with all radical prostatectomies. CONCLUSIONS: Discrepancies exist in practice patterns between urologists as well as inconsistencies in logic within individuals. There is little variation between individuals in different practice settings. Our results reflect the often confusing and conflicting data published during the last decade. PMID- 9334618 TI - Racial differences in operating characteristics of prostate cancer screening tests. AB - PURPOSE: We evaluated racial differences in the operating characteristics of prostate specific antigen (PSA) and digital rectal examination as screening tests for early detection of prostate cancer. MATERIALS AND METHODS: We screened 18,527 white and 949 black men 50 years old or older using serum PSA measurement and digital rectal examination. We recommended biopsies if either test was suspicious for cancer. For PSA greater than 4.0 ng./ml. and rectal examination we calculated relative sensitivity (percentage of men with cancer who had a positive test), specificity (percentage of men without cancer who had a negative test) and positive predictive value (percentage of men with a positive test in whom cancer was detected) for the prediction of prostate cancer stratified by race. RESULTS: In white and black men PSA greater than 4.0 ng./ml. detected more cancers than rectal examination (p < 0.002) with a trend for a greater increase in sensitivity in black men. PSA was associated with fewer false-positives than rectal examination in white (p < 0.0001) but not in black (p > 0.05) men. Positive predictive value for prostate cancer of PSA and rectal examination was greater in black than in white men (48 versus 34 and 38 versus 22%, respectively). CONCLUSIONS: PSA detects more cancers than rectal examination in both races, although this advantage is more pronounced in black men. In white men PSA yielded fewer false-positive results than rectal examination. However, PSA had more false positive results than rectal examination in black men. Cancer was detected in a higher percentage of black men with PSA greater than 4.0 ng./ml. and, therefore, the risk of cancer associated with PSA greater than 4.0 ng./ml. differs by race. In a screening setting the widely accepted 25 to 30% positive predictive value for PSA greater than 4.0 ng./ml. may only apply to white men. A higher risk estimate of 36 to 60% is more accurate in black men. PMID- 9334619 TI - Influence of hepatic function on serum levels of prostate specific antigen. AB - PURPOSE: Recent studies have suggested that the primary site of metabolism for prostate specific antigen (PSA) is the liver. We evaluated men undergoing liver transplantation to determine whether chronic hepatic insufficiency affected serum PSA levels and whether improved hepatic function altered serum PSA levels. MATERIALS AND METHODS: Ten men with a mean age of 46 years (range 23 to 67) undergoing liver transplantation were evaluated. Liver function tests, including serum bilirubin, serum glutamic-oxaloacetic transaminase and serum glutamic pyruvic transaminase, as well as serum PSA were determined 1 day before and a mean of 12.6 months (range 4 to 18) after transplantation. RESULTS: Serum bilirubin and serum glutamic-oxaloacetic transaminase declined significantly after liver transplantation. There was no difference in mean serum PSA levels before and after liver transplantation. CONCLUSIONS: Our results suggest that severe hepatic dysfunction does not significantly alter the serum concentration of PSA. These data, combined with recent investigations demonstrating an intrahepatic mechanism for PSA elimination, suggest that the liver has a significant reserve to metabolize the relatively small quantities of PSA in the circulation. PMID- 9334620 TI - Prostate specific antigen reverse transcriptase-polymerase chain reaction assay in preoperative staging of prostate cancer. AB - PURPOSE: Extracapsular extension of prostate cancer occurs in a significant number of men believed to have clinically localized disease. We report the ability of the reverse transcriptase-polymerase chain reaction (RT-PCR) assay to predict preoperatively the pathological stage of cases of clinically localized prostate cancer. MATERIALS AND METHODS: Since October 1994, 82 consecutive men with clinically localized prostate cancer had a venous blood RT-PCR assessment before radical retropubic prostatectomy. The extracted ribonucleic acid was reverse transcribed, amplified and the amplicon identity confirmed by prostate specific antigen (PSA) directed probe hybridization. An additional 31 patients were enrolled to provide appropriate positive (T + Nx/1M2) and negative (human female and benign prostatic hyperplasia) controls. Histological examination of the entire prostatectomy specimen was performed. RESULTS: Positive RT-PCR assay results correlated significantly with skeletal metastases and elevated levels of serum PSA but they did not significantly improve our ability to identify prospectively patients with extracapsular extension over traditional predictors (serum PSA, Gleason score). CONCLUSIONS: The role of molecular techniques in prostate cancer evaluation and prognosis continues to emerge. However, in our study we demonstrate no significant advantage in preoperative staging of prostate cancer using RT-PCR assay with PSA primers. PMID- 9334621 TI - Pretreatment prostate specific antigen doubling times: use in patients before radical prostatectomy. AB - PURPOSE: We determined whether pre-radical prostatectomy prostate specific antigen (PSA) doubling time could predict pathological stage at radical prostatectomy or PSA failure postoperatively. We also sought to compare PSA doubling times from men with prostate cancer treated with radical prostatectomy to a group treated with radiation therapy. MATERIALS AND METHODS: Detailed followup was available for 150 patients with clinically localized prostate cancer who underwent radical prostatectomy from January 1993 to August 1995. PSA doubling time was calculated for all patients with 3 or more pre-radical prostatectomy PSA levels using linear regression. We assessed the association between PSA doubling time and PSA failure, pathologic stage at radical prostatectomy, final PSA before treatment and Gleason score. We compared our PSA doubling time values and distribution to a published series of patients with prostate cancer who had undergone radiation therapy. RESULTS: A total of 56 patients had 3 or more PSA values before treatment. Median followup was 17.3 months. PSA doubling time did not correlate with PSA failure, final PSA or Gleason score, but it did with pathological stage at radical prostatectomy (p = 0.0035 for positive margins, p = 0.025 for positive seminal vesicles). Our PSA doubling time and PSA failure rates did not differ from the radiation therapy population with similar followup times. CONCLUSIONS: Although studies from the radiation literature have shown PSA doubling time to be useful in predicting PSA failure after treatment for prostate cancer, our results do not confirm this finding. We did find a correlation with pathologic stage at radical prostatectomy, and so longer followup with more patients may confirm this in the future. We also found no significant differences in PSA doubling time between our patients and a group treated with radiation. At least for this parameter, patients with prostate cancer referred for radical prostatectomy and radiation therapy may be similar. PMID- 9334622 TI - The prognostic value of pretreatment expression of androgen receptor and bcl-2 in hormonally treated prostate cancer patients. AB - PURPOSE: We determined the prognostic value of oncoprotein bcl-2 and androgen receptor expression in pretreatment transurethral resection specimens of hormonally treated prostate cancer patients. MATERIALS AND METHODS: A total of 68 pretreatment transurethral resection specimens, 30 radical prostatectomy specimens and 21 palliative transurethral resection specimens with androgen independent prostate cancer was stained with a monoclonal antibody against bcl-2. Androgen receptor immunohistochemistry was performed on pretreatment transurethral resection specimens only. Results were scored semiquantitatively and were correlated with tumor stage and grade and with the occurrence of clinical progression or tumor related death. RESULTS: Bcl-2 expression by adenocarcinoma cells was found in 32, 17 and 24% of pretreatment transurethral resection, radical prostatectomy and palliative transurethral resection specimens, respectively. The bcl-2 scores did not correlate with tumor stage or grade. Androgen receptor was expressed in 88% of pretreatment transurethral resection specimens. Androgen receptor scores were marginally related to tumor grade, but not to tumor stage. A prognostic value of bcl-2 or androgen receptor in pretreatment transurethral resection specimens was not found. When a combined bcl-2/androgen receptor score was used, this parameter was an independent prognostic marker to predict clinical progression with Gleason grade and stage classification. Gleason grade was the only independent prognostic marker to predict tumor related death. CONCLUSIONS: The expression of bcl-2 and androgen receptor in pretreatment prostate cancer specimens is not related to the prognosis of hormonally treated prostate cancer. Bcl-2 expression is not increased in endocrine therapy resistant prostate cancer. Surprisingly, a combined bcl-2/androgen receptor score acts as an independent prognosticator for clinical progression. PMID- 9334624 TI - Laparoscopic pelvic lymph node dissection for prostate cancer: comparison of the extended and modified techniques. AB - PURPOSE: We compared the results of extended (obturator, hypogastric, common and external iliac nodes) to modified (obturator and hypogastric nodes only) laparoscopic pelvic lymph node dissection in patients with clinically localized prostate cancer. MATERIALS AND METHODS: A total of 189 patients with stage T1 to T3 prostate cancer underwent modified (150) or extended (39) laparoscopic pelvic lymph node dissection for pelvic nodal assessment before definitive treatment. RESULTS: Twice as many lymph nodes were removed via extended than modified laparoscopic pelvic lymph node dissection (mean 17:8 versus 9.3). The overall positivity rate was 23 of 189 lymph nodes (12.2%), including 14 of 150 (7.3%) for modified and 9 of 39 (23.1%) for extended dissection (p = 0.02). Two patients (22%) who underwent extended dissection had positive lymph nodes in the external iliac area. Patients who presented with the high risk features of prostate specific antigen (PSA) greater than 20 ng./ml., Gleason score 7 or greater, or stage T2b disease or greater had a 26.5% (p = 0.0002), 22% (p = 0.0006) or 16.4% (p = 0.003) likelihood of positive lymph nodes, respectively. For extended versus modified laparoscopic pelvic lymph node dissection node positivity in high risk patients was 27% versus 18.8% (p = 0.4), 30 versus 26.4% (p = 0.8) and 25.4 versus 14.6% (p = 0.17) for Gleason score 7 or greater, PSA greater than 20 ng./ml. and disease stage T2b to T3a, respectively. Patients who underwent the extended procedure had a higher complication rate (35.9 versus 2%, p < 0.0001). No laparotomy was required. CONCLUSIONS: Despite yielding a 2-fold higher node count and higher node positivity rate, extended laparoscopic pelvic lymph node dissection offers no advantage over modified laparoscopic pelvic lymph node dissection for diagnosing positive lymph nodes when results are analyzed by prognostic factors. The extended procedure is associated with a much higher complication rate. In patients with the high risk features of PSA greater than 20 ng./ml., Gleason score 7 or greater and stage T2b to T3a disease modified laparoscopic pelvic lymph node dissection can be performed safely and effectively to help identify those who may benefit most from curative therapy. PMID- 9334625 TI - Transitional cell carcinoma in situ of the seminal vesicles: 8 cases with discussion of pathogenesis, and clinical and biological implications. AB - PURPOSE: Mucosal migration of transitional cell carcinoma in situ is a potential mechanism for multifocal lower tract disease. MATERIALS AND METHODS: The clinical course and pathological studies of 8 cases of carcinoma in situ are reviewed in detail. RESULTS: The pattern of disease of carcinoma in situ of seminal vesicle provides circumstantial evidence for mucosal migration of cancer from a bladder or prostatic urethral origin. CONCLUSIONS: A monoclonal origin of bladder cancer combined with mucosal spread of carcinoma in situ suggests that incomplete destruction of carcinoma in situ may adversely affect long-term results by permitting extension into the distal ureters, prostatic duct or seminal vesicles. Protracted intravesical treatment of carcinoma in situ without complete elimination of the disease allows the natural history of mucosal spread to become evident. PMID- 9334623 TI - Use of repeat sextant and transition zone biopsies for assessing extent of prostate cancer. AB - PURPOSE: Little is known why certain prostate cancers are missed on biopsy. In patients with a needle biopsy diagnosis of cancer it is also unknown whether repeat needle biopsy provides useful information to predict extent of disease. MATERIALS AND METHODS: In the pathology laboratory we performed sextant and transition zone needle biopsies on 193 radical prostatectomy specimens from men with nonpalpable cancer detected on needle biopsy (stage T1c) using an 18 gauge biopsy gun. Radical prostatectomy specimens were then serially sectioned, totally embedded, mapped and staged. RESULTS: The transition zone biopsy by itself was positive in only 2.1% of cases, demonstrating the lack of usefulness for this particular biopsy. Despite cancer on preoperative needle biopsy in all cases, 31% showed no cancer on repeat sextant transition zone biopsy. In a multivariate analysis (variables included radical prostatectomy tumor volume, radical prostatectomy tumor location, prostate gland size and radical prostatectomy grade) decreased tumor volume (p < 0.0001), increased gland size (p = 0.001), and decreased radical prostatectomy grade (p = 0.013) were each independent predictors of absence of tumor on repeat biopsy. A lack of cancer on repeat biopsy correlated with pathological stage: 90% of cases without cancer on repeat biopsy were organ confined versus 66% for cases with a single less than 3 mm. focus of cancer on repeat biopsy versus 58% for cases with more cancer on repeat biopsy. Of 38 men with a preoperative needle biopsy showing less than 3 mm. of cancer on 1 core that was not high grade and with prostate specific antigen 10 or less (men for whom urologists are most likely to repeat biopsy) the presence of cancer on repeat biopsy also correlated with extent of disease at radical prostatectomy. However, of these 38 men 6 of 16 with no cancer on repeat biopsy had moderate tumor (4 with organ confined Gleason score 5 to 6, tumor volume 0.79 to 4.5 cc; 1 with organ confined Gleason score 7, tumor volume 0.18 cc; and with 1 established penetration Gleason score 6, tumor volume 0.53) at radical prostatectomy. CONCLUSIONS: Although absence of cancer on repeat biopsy correlates with various parameters of extent of disease, there is significant overlap for the individual patient. This study also demonstrates the limits of sextant needle biopsy to evaluate tumor status in patients who elect watchful waiting or less invasive forms of therapy (cryotherapy, interstitial radiotherapy). PMID- 9334626 TI - Comparison of urine collection methods for evaluating urinary nuclear matrix protein, NMP22, as a tumor marker. AB - PURPOSE: The nuclear matrix protein, NMP22, has been shown to be a useful tumor marker for identifying patients with a high likelihood of rapid recurrence of transitional cell carcinoma of the urinary tract after surgical treatment. Currently measurement of NMP22 involves 3 urine voids collected during a 24-hour period, which are pooled and assayed as a single sample. This study was performed to determine whether any single void would yield similar results to the pooled 3 void sample. MATERIALS AND METHODS: A total of 2,018 urine samples (3 voids per sample) was included in the study. All analyses were performed using the nonparametric Wilcoxon signed rank test for matched pairs. RESULTS: Analysis showed that the NMP22 level of a single void collected between midnight and noon was similar to the NMP22 level of the pooled 3-void sample. Receiver operating characteristics curves of the midnight-to-noon single void and the pooled 3-void sample were similar for predicting recurrence postoperatively in patients with urinary tract transitional cell carcinoma. CONCLUSIONS: One void collected between midnight and noon compares favorably with the current 3-void collection method for determining NMP22 levels in urine. PMID- 9334627 TI - Failure of an acucise balloon device to inflate resulting in treatment failure. PMID- 9334628 TI - Polypoid arteriovenous malformation of the ureter. PMID- 9334629 TI - The gastroileal neobladder for renal insufficiency. PMID- 9334630 TI - Acute urinary retention due to an iatrogenic bladder diverticulum. PMID- 9334631 TI - Bilateral keratinizing desquamative squamous metaplasia of the upper urinary tract. PMID- 9334632 TI - Ventral bladder hernia following vesica percutaneous bladder neck suspension for stress urinary incontinence. PMID- 9334633 TI - Leiomyoma of female urethra causing acute urinary retention and acute renal failure. PMID- 9334634 TI - Primary adenosquamous cell carcinoma of the male distal urethra: magnetic resonance imaging using a circular surface coil. PMID- 9334635 TI - Ureteral Stones Clinical Guidelines Panel summary report on the management of ureteral calculi. The American Urological Association. AB - PURPOSE: The American Urological Association convened the Ureteral Stones Clinical Guidelines Panel to analyze the literature regarding available methods for treating ureteral calculi and to make practice policy recommendations based on the treatment outcomes data. MATERIALS AND METHODS: The panel searched the MEDLINE data base for all articles related to ureteral calculi published from 1966 to January 1996. Outcomes data were extracted from articles accepted after panel review. The data were then meta-analyzed to produce outcome estimates for alternative treatments of ureteral calculi. RESULTS: The data indicate that up to 98% of stones less than 0.5 cm. in diameter, especially in the distal ureter, will pass spontaneously. Shock wave lithotripsy is recommended as first line treatment for most patients with stones 1 cm. or less in the proximal ureter. Shock wave lithotripsy and ureteroscopy are acceptable treatment choices for stones 1 cm. or less in the distal ureter. CONCLUSIONS: Most ureteral stones will pass spontaneously. Those that do not can be removed by either shock wave lithotripsy or ureteroscopy. Traditional blind basket extraction, without fluoroscopic control and guide wires, is not recommended. Open surgery is appropriate as a salvage procedure or in certain unusual circumstances. PMID- 9334636 TI - Re: The importance of the depth of invasion in stage T1 bladder carcinoma: a prospective cohort study. PMID- 9334637 TI - Re: Giant scrotal lipomatosis. PMID- 9334638 TI - Re: Laser prostatectomy performed with right angle firing neodymium: YAG laser fiber at 40 watts power setting. PMID- 9334639 TI - Wilms tumor in a prenatally diagnosed multicystic kidney. PMID- 9334640 TI - Laparoscopic heminephroureterectomy in pediatric patients. AB - PURPOSE: An increasing number of operative procedures in pediatric urology can be performed by laparoscopy. We report our experience with laparoscopic heminephroureterectomy, which is a typical operation in pediatric patients. MATERIALS AND METHODS: Laparoscopic heminephroureterectomy was performed in 14 consecutive children. In 12 cases 7 upper renal poles were removed for ectopic refluxing megaureter and obstructive ureterocele in 5 and 2, respectively. In 5 children lower poles were destroyed by reflux nephropathy. In 2 children laparoscopic upper pole heminephroureterectomy for obstructive ureterocele was combined with a Pfannenstiel incision for reimplantation of the refluxing lower pole ureter. RESULTS: All operations were completed as planned. Operative time was 180 to 330 minutes (mean 222) in group 1 and 345 to 510 (mean 427) in group 2. Blood loss was minimal (10 to 30 ml.) and there were no intraoperative or postoperative complications. Mean postoperative hospital stay in groups 1 and 2 was 4.4 and 7.5 days, respectively. CONCLUSIONS: Laparoscopic heminephroureterectomy in children is feasible and associated with minimal blood loss, low morbidity and a low complication rate. The disadvantage is the long operative time. This technically demanding procedure should be performed only at specialized centers. PMID- 9334641 TI - Ectopic vaginal insertion of an upper pole ureter: demonstration by special sequences of magnetic resonance imaging. PMID- 9334642 TI - Bilateral single ureteral ectopia: difficulty attaining continence using standard bladder neck repair. AB - PURPOSE: We reviewed the surgical results of the management of bilateral single ureteral ectopia, a rare congenital cause of severe urinary incontinence. MATERIALS AND METHODS: We reviewed the records of 6 girls and 1 male infant who presented to 1 institution with this diagnosis in a 10-year period. RESULTS: All patients were incontinent and 3 had undergone ureteral reimplantation as an initial procedure with persistent postoperative wetting. Of the 5 patients who underwent a total of 8 attempts at increasing bladder outlet resistance, including 3 Young-Dees-Leadbetter, 2 Kropp, 1 Stamey, 1 Burch and 1 pubovaginal sling procedure, 2 also underwent simultaneous bladder augmentation to increase bladder capacity. However, none of these children had satisfactory continence after the continence procedure. Three of these patients who subsequently underwent appendicovesicostomy with bladder neck closure are continent. The 2 remaining patients underwent initial appendicovesicostomy with bladder neck closure and augmentation, and they are also continent. CONCLUSIONS: In our series total day and nighttime continence was only achieved by bladder neck closure, appendicovesicostomy and augmentation. Attempts at increasing bladder outlet resistance in patients with bilateral single ectopic ureters led to suboptimal rates of success even when adequate bladder capacity had been ensured by simultaneous augmentation. PMID- 9334643 TI - Lack of distant migration after injection of a 125iodine labeled dextranomer based implant into the rabbit bladder. AB - PURPOSE: In recent years endoscopic treatment of stress incontinence and vesicoureteral reflux has been introduced. Reports of possible particle migration of the injected material to distant organs in humans and experimental animals have led to a search for biological nonmigration products. An implant found to have a good clinical effect in these conditions is dextranomer in hyaluronan. We performed this study in rabbits to investigate the possible migration of dextranomer particles. MATERIALS AND METHODS: 125Iodine labeled dextranomer particles were injected into the submucosal space of rabbit bladders, and samples of blood and various tissues were examined for radioactivity at scheduled intervals during a 28-day period. Furthermore, whole body autoradiography was performed 1 day, and 1 and 4 weeks after injection. RESULTS: Radioactivity was found in blood samples and in all tissues but it remained at the background activity level except in the thyroid, where uptake representing free 125iodine was detected. In the bladder 41 and 45% of the injected dose remained within the bladder wall 1 day and 4 weeks, respectively, after injection. The remainder of the dose probably disappeared from the bladder wall by leakage into the urine shortly after deposition, as indicated by the finding of 10-fold higher urine radioactivity levels at day 1 than at day 28 after injection. CONCLUSIONS: No distant migration of dextranomer particles occurs after submucosal injection of such an implant in the rabbit bladder wall. PMID- 9334644 TI - Results of urinary tract reconstruction in boys with end stage bladders resulting from obstructive uropathy. AB - PURPOSE: The records of 17 boys who underwent reconstruction of the lower urinary tract because of end stage bladders resulting from obstructive uropathy were reviewed to evaluate the degree to which they void and factors that favorably impacted outcome. MATERIALS AND METHODS: A total of 17 boys with end stage bladders resulting from obstructive uropathy, including posterior urethral valves in 15 and obstructing ureteroceles in 2, underwent reconstruction of the urinary tract. Procedures consisted of augmentation (autoaugmentation in 3, ileocystoplasty in 2, ileocecal cystoplasty in 1 and colocystoplasty in 10), an ileal ureter in 2, bladder neck revision in 15 and appendicovesicostomy in 8. RESULTS: All patients achieved a low pressure reservoir of adequate volume with stable or improved urinary tracts. A total of 13 patients voided sufficiently well to maintain a favorable life-style, including 5 who were completely catheter free. With respect to the variables involved in reconstruction, bladder neck revision seemed to correlate best with a good outcome. CONCLUSIONS: In most cases end stage bladders in boys with obstructive uropathy can be reconstructed not only to protect the urinary system, but to preserve some degree of voiding potential as well. Bladder neck revision appears to be particularly helpful in achieving the latter goal. When voiding is inadequate, appendicovesicostomy provides easy access to the bladder for intermittent catheterization. PMID- 9334645 TI - A serious circumcision complication: penile shaft amputation and a new reattachment technique with a successful outcome. PMID- 9334646 TI - Benefits of laparoscopy and the Jones technique for the nonpalpable testis. AB - PURPOSE: We report on the role of diagnostic laparoscopy combined with the Jones suprainguinal approach to orchiopexy in the treatment of abdominal testes. MATERIALS AND METHODS: A retrospective review was done of 209 boys with 265 cryptorchid testes who underwent orchiopexy between January 1994 and March 1996. A subset of patients with nonpalpable testis underwent diagnostic laparoscopy and according to the laparoscopic findings either standard inguinal or suprainguinal extraperitoneal orchiopexy. RESULTS: Of 209 patients 47 had 63 impalpable testes. Laparoscopy was done on 31 patients (42 testes). Findings at laparoscopy revealed 29 (69%) abdominal, 7 (17%) intracanalicular and 6 (14%) vanishing testes. A satisfactory result with scrotal position of the testis and no atrophy was obtained in 18 of 19 patients operated via the Jones approach combined with diagnostic laparoscopy. The only patient with unsatisfactory result underwent ligation of the spermatic vessels at the time of operation. In contrast, satisfactory results were achieved for only 7 of 10 intra-abdominal testes after diagnostic laparoscopy and standard inguinal orchiopexy despite universal spermatic vessel ligation. Of these 10 testes 2 ended in an inguinal location and 1 atrophied. CONCLUSIONS: Laparoscopy was helpful in determining surgical approach in most cases. The suprainguinal approach allowed most abdominal testes to be placed in the scrotum without vascular ligation. PMID- 9334648 TI - Will laparoscopic orchiopexy replace open surgery for the nonpalpable undescended testis? PMID- 9334647 TI - The value of 2-step laparoscopic Fowler-Stephens orchiopexy for intra-abdominal testes. AB - PURPOSE: We report our experience in the treatment of high intra-abdominal testis with a complete laparoscopic 2-stage Fowler-Stephens procedure with associated transperitoneal closure of the internal ring in pediatric patients. MATERIALS AND METHODS: Between 1990 and 1997, 100 boys with 105 nonpalpable testes underwent laparoscopy. Laparoscopy showed intra-abdominal testis in 40 cases. In 5 cases when the testis was just proximal to the internal ring, we performed standard orchiopexy. In 35 cases with the testis in the high intra-abdominal position, we performed the Fowler-Stephens procedure with the first stage performed laparoscopically. To date, all 35 testis have undergone the second phase after 6 to 12 months (2 by open technique, 33 by laparoscopy). The last 33 patients underwent the second phase of the 2-stage Fowler-Stephens procedure by laparoscopy with associated video surgical transperitoneal closure of internal ring. RESULTS: All testes were successfully placed in the scrotum. At a mean 30 months of followup, with clinical examination, ultrasonography and comparative colorimetric echo Doppler study, all testes were viable in the scrotum, except for 1 that became atrophic 2 months after the second open phase of 2-stage Fowler Stephens technique. CONCLUSIONS: Our early results suggest that the 2-stage Fowler-Stephens procedure, performed completely using laparoscopy, is a feasible technique for treating high intra-abdominal testis. PMID- 9334649 TI - Bilateral testicular torsion in the neonatal period. PMID- 9334650 TI - Meconium filled hydrocele sacs as a cause of acute scrotum in a newborn. PMID- 9334651 TI - Alterations in endothelin B receptor sites in cavernosal tissue of diabetic rabbits: potential relevance to the pathogenesis of erectile dysfunction. AB - PURPOSE: Diabetes Mellitus (DM) is a major risk factor for erectile dysfunction in both patients and animal models. The pathogenesis of this dysfunction has not been fully elucidated. However, alterations in the synthesis of a number of vasoactive compounds, such as nitric oxide (NO) and prostacyclin (PGI2), have been reported in various diabetic tissues. The interaction between NO, PGI2 and endothelin-1 (ET-1), a powerful vasoconstrictor and smooth muscle cell mitogen, is thought to be important in maintaining vascular tone and the erectile process. We investigated the density and distribution of ET-1 and endothelin receptor subtypes in cavernosal tissue and assessed any changes brought about by DM in a rabbit model. MATERIALS AND METHODS: DM was induced in New Zealand White rabbits using alloxan. Penises were excised from the diabetic rabbits three months (n = 6) and six months (n = 6) after the induction of DM. Low and high resolution autoradiography was performed using radioligands for ET-1, endothelin A (ETA) and endothelin B (ETB) receptors and were analyzed densitometrically. The results were compared with those from six age-matched healthy control rabbits for each group. Immunohistochemical localization of ET-1 immunoreactivity was also performed, together with ultrastructural evaluation of the corpus cavernosum. RESULTS: ET-1, ETA and ETB receptor binding sites were primarily localized to the smooth muscle cells of the corpus cavernosum and the endothelium lining the cavernosal spaces. A significant increase in ETB receptor binding sites was found only in cavernosal tissue six months after induction of DM, when compared with age-matched healthy controls. These receptor changes were accompanied by ultrastructural changes in the corpus cavernosum indicative of an early, atherosclerosis-like process. CONCLUSIONS: The autoradiographic and immunohistochemical findings in this study suggest that ET-1 may have a role in the pathophysiology of diabetic ED. This peptide may be released in an autocrine fashion causing cavernosal smooth muscle cell (CSMC) contraction and/or proliferation. PMID- 9334653 TI - Effect of cryptorchidism on testicular histology in a naturally cryptorchid animal model. AB - PURPOSE: To determine the effect of naturally occurring cryptorchidism on testicular histology in both the cryptorchid and normally descended testis from birth to adulthood using the LE/ORL rat model. MATERIALS AND METHODS: Testicular histology was assessed using established morphometric measures in bilaterally descended (BD), unilaterally descended (UD), bilaterally cryptorchid (BC) and unilaterally cryptorchid (UC) testis at days 15, 22, 30, 45 and 60 of age. Testicular mass was also measured at these times. RESULTS: Changes in testicular histology in the BC and UC testes were not noted on or prior to day 30 of age. Significant changes were noted by day 45 of age and continued into adulthood at day 60 of age. There were no histological abnormalities noted in the UD and BD groups. CONCLUSIONS: Since histological changes seen in this animal model occur after the time of testicular descent (day 28 of age), we hypothesize that these changes are due to an abnormal anatomical position of the testis as opposed to an inherent testicular defect in the LE/ORL rat. This hypothesis is supported by the fact that no histological differences were noted between the scrotal testes of unilaterally cryptorchid animals and bilaterally descended control animals. PMID- 9334652 TI - Evidence of adenosine 5'-triphosphate release from nerve and P2x-purinoceptor mediated contraction during electrical stimulation of rat urinary bladder smooth muscle. AB - PURPOSE: We provided direct evidence for the existence of purinergic innervation in the rat urinary bladder. MATERIALS AND METHODS: The non-adrenergic non cholinergic (NANC) innervation was studied in 4-month-old Wistar rats. Electric field stimulation (EFS) of the detrusor muscle strips in the presence of four autonomic blockers (atropine 10(-6) M, guanethidine 10(-6) M, phentolamine 10(-6) M and propranolol 10(-6) M) showed NANC contractions accounted for about 50% of the maximum contractile response. The adenyl purines released from nerves by EFS were detected by HPLC after conversion to ethenopurines. The amount of total purine released was frequency-dependent and could be totally suppressed by tetradotoxin (10(-6) M). The amount of ATP released was significantly greater than those for ADP, AMP and adenosine (p < 0.05, n = 4). Desensitization induced by alpha, beta-MeATP (10(-6) to 10(-4) M), a P2x receptor agonist, reduced the NANC contraction. In addition, the NANC contraction was also abolished by P2 receptor blocker suramin (10(-4) to 10(-3) M) and P2x receptor blocker PPADS (10( 5) to 10(-4) M.). CONCLUSION: The results of the present study give evidence to support purinergic nerve-mediated bladder smooth muscle contractions in the rat. Among the purine nucleotides, ATP is the dominant purinergic neurotransmitter released and P2x receptor activation is responsible for the NANC contractile response. PMID- 9334654 TI - Concentrations of specific epithelial growth factors in the urine of interstitial cystitis patients and controls. AB - PURPOSE: Interstitial cystitis (IC) is a chronic bladder disease for which the etiology is unknown. Because the bladder epithelium is often abnormal in IC, we determined whether the levels of specific urine growth factors postulated to be important for bladder epithelial proliferation are altered in IC. MATERIALS AND METHODS: ELISAs were used to determine levels of epidermal growth factor (EGF), insulin-like growth factor 1 (IGF1), insulin-like growth factor binding protein 3 (IGFBP3), and heparin binding epidermal growth factor-like growth factor (HB-EGF) in urine specimens from women with IC, asymptomatic women without bladder disease, and women with bacterial cystitis. RESULTS: Urine HB-EGF levels were specifically and significantly decreased in IC patients as compared to asymptomatic controls or patients with bacterial cystitis, whether expressed as concentration (amount per volume of urine) or the amount relative to urine creatinine in each specimen. In contrast, urine EGF, IGF1, and IGFBP3 levels were all significantly elevated in IC patients compared to asymptomatic controls. Further, the amounts of urine EGF and IGF1 were also elevated in IC patients as compared to patients with bacterial cystitis, and urine IGFBP3 levels were significantly elevated when expressed per milligram of urine creatinine. CONCLUSIONS: These findings indicate that complex changes in the levels of urine epithelial cell growth factors (EGF, IGF1, and HB-EGF) and a growth factor binding protein (IGFBP3) are associated with IC. While EGF, IGF1, and IGFBP3 levels are either the same or increased in the urine of IC patients as compared to patients with bacterial cystitis or asymptomatic controls, HB-EGF levels are significantly decreased in the urine of IC patients. Understanding the reasons for these changes may lead to understanding the pathogenesis of this disorder. PMID- 9334655 TI - DMSO: effect on bladder afferent neurons and nitric oxide release. AB - PURPOSE: Interstitial cystitis (IC), a chronic disorder of the urinary bladder, is characterized by increased voiding frequency, urgency and pain. Patients with IC also exhibit reduced urinary nitric oxide synthase activity. Intravesical administration of dimethyl sulfoxide (DMSO) has been used to provide symptomatic relief in patients with IC. The present experiments were undertaken to determine if intravesical DMSO affects neural pathways involved in bladder function in the rat and if DMSO can influence the release of nitric oxide in the bladder or from afferent neurons. MATERIALS AND METHODS: The effects of intravesical DMSO (10% solution in saline) on reflex bladder activity, firing on bladder nerves and c fos gene expression in spinal neurons was examined in urethane anesthetized female Wistar rats. The effect of DMSO (1-10%) on nitric oxide release from urinary bladder strips or acutely dissociated dorsal root ganglion cells was monitored in vitro with a porphyrinic microsensor. RESULTS: DMSO acutely increased reflex firing of pelvic nerve efferent axons, decreased bladder capacity and also increased neuronal c-fos expression in spinal cord regions that exhibit c-fos expression after chemical activation of capsaicin-sensitive bladder afferents. DMSO, like capsaicin, also directly released nitric oxide (NO) from both dissociated dorsal root ganglion neurons and from isolated strips of urinary bladder. CONCLUSIONS: These results suggest that DMSO induced stimulation of bladder afferent pathways and NO release from afferent neurons may be a reflection of the initial event in the desensitization of nociceptive pathways in the lower urinary tract (LUT). PMID- 9334656 TI - Association of in vitro growth potential of urinary exfoliated cells with tumor localization and intraluminal recurrence rates of urothelial cancers. AB - PURPOSE: One problem in the treatment of urothelial cancers, in particular of upper urinary tract cancers, is multifocal synchronous and/or metachronous tumor development in the heterotopic urothelium. We investigated the abilities of exfoliated cells in the urine of patients with urothelial cancers to colonize and proliferate in vitro. MATERIALS AND METHODS: Short-term cultures of 129 urine samples obtained from patients with urothelial cancers and 53 samples from healthy volunteers were compared as to the growth potential of urinary-exfoliated cells and clinico-pathological features, in particular tumor localization and evidence of intraluminal tumor recurrence. RESULTS: Primary cell outgrowth occurred in 112 (86.8%) of the 129 cell cultures from the patients and in 29 (54.7%) of the 53 from the healthy volunteers (p < 0.0001). "Sufficient cell growth" (more than 10(5) cells per flask) was obtained for 77 (59.7%) of the 129 cultures from patients and 7 (13.2%) of the 53 from the healthy volunteers (p < 0.0001). In terms of tumor localization, the rate of sufficient cell growth was significantly higher for patients with upper urinary tract tumor (21/25, 84.0%) than for those with bladder tumor (56/104, 53.8%) (p = 0.0062). Proportional hazard regression analysis showed that only the growth potential of the urinary exfoliated cells was significantly predictive of intravesical tumor recurrence in patients with superficial bladder tumors (p = 0.011). CONCLUSIONS: The potentials for the colonization and proliferation of urinary-exfoliated cells are associated with intraluminal multifocal tumor recurrence of urothelial cancers. PMID- 9334657 TI - Alterations of the metastasis suppressor gene nm23 and the proto-oncogene c-myc in human testicular germ cell tumors. AB - The putative metastasis suppressor genes nm23-H1, nm23-H2 and the c-myc proto oncogene were investigated in testicular germ cell tumors (GCTs) using Southern and Northern blotting as well as semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and single strand conformation polymorphism (SSCP) analysis. When studying Bgl II RFLPs, allelic losses of the nm23 gene were found in 3/12 (25%) informative tumors, and all 3 had lymph node and/or distant metastases. A 2 to 7 fold nm23 mRNA overexpression was found in 22/34 (64.7%) tumors examined. RT-PCR revealed that this phenomenon is mainly a consequence of nm23-H2 overexpression. Overexpression of both the H1 and the H2 gene was predominantly found in the seminoma subtype and was not associated with tumor stage. Only 1/25 tumors, a seminoma with distant metastases, had a point mutation in the coding region of the nm23-H2 gene as demonstrated by SSCP analysis. None of the 8 seminomas and only 1/13 non-seminomas had c-myc overexpression. No abnormalities of the c-myc gene could be detected on the DNA level. Despite the fact that in previous investigations nm23-H2 was demonstrated to be a putative transcription factor for c-myc, no coexpression of c-myc and nm23-H2 was found by quantitative RT-PCR in this study. PMID- 9334697 TI - Wrong kind of chemokine research. PMID- 9334658 TI - Simultaneous injection of round spermatid nuclei from mice and hamster oscillogen can initiate the normal development of mouse embryos. AB - PURPOSE: In the mouse, mature oocytes that have been injected with spermatid nuclei failed to become activated. Additional stimulation is required to trigger activation of the oocytes that leads to embryo development. This study was performed to determine whether simultaneous injection of mouse round spermatids and hamster oscillogen can fertilize oocytes and contribute to normal embryo development. MATERIALS AND METHODS: We simultaneously injected mouse oocytes with mouse round spermatid nuclei and a preparation of oocyte-activating protein (oscillogen) from hamster spermatozoa and compared the results with those of injection of testicular spermatozoa. RESULTS: The incidence of normal fertilization and embryo development after simultaneous injection of round spermatid nuclei and hamster oscillogen was not significantly different from that after the injection of testicular spermatozoa. Moreover, the rate of development of two-cell embryos to term did not differ very much between the two groups of oocytes. CONCLUSIONS: Simultaneous injection of round spermatid nuclei and oscillogen appears to be an ideal method for successful activation of oocytes by spermatid nuclei in vitro. PMID- 9334698 TI - Are anti-HIV drugs an effective treatment? PMID- 9334699 TI - CCR2 chemokine receptor and AIDS progression. PMID- 9334700 TI - Italian mental hospitals closer to closing? PMID- 9334701 TI - AIDS vaccine trial. Publicity stunt or genuine attempt at progress? PMID- 9334702 TI - From human rights to the human genome--stopping biopiracy. PMID- 9334703 TI - US government pays hospitals not to train doctors... PMID- 9334704 TI - Controversial muscular dystrophy therapy goes to court. PMID- 9334705 TI - Fetal tissue and increased funding for Parkinson's disease. PMID- 9334706 TI - 1997 Albert Lasker Award for Clinical Research. Clinical research and the human condition: moving from observation to practice. PMID- 9334707 TI - 1997 Albert Lasker Award for Special Achievement in Medical Science. Observations over 50 years concerning intestinal polyposis, Marfan syndrome and achondroplasia. PMID- 9334708 TI - 1997 Albert Lasker Award for Basic Medical Research. Control of gene transcription: an outline. PMID- 9334709 TI - At the scene of ischemic brain injury: is PARP a perp? PMID- 9334710 TI - A chemokine trap for HIV co-receptors. PMID- 9334711 TI - Gastrogenomic delights: a movable feast. PMID- 9334712 TI - A coat of many complements. PMID- 9334713 TI - Diabetes in midlife: planting genetic time bombs. PMID- 9334714 TI - Is breast cancer a potential side effect of GH treatment? PMID- 9334715 TI - bFGF and tumor angiogenesis--back in the limelight? PMID- 9334716 TI - A new role for an old actor. PMID- 9334717 TI - Viral damage and the breakdown of self-tolerance. PMID- 9334718 TI - A tail retold. PMID- 9334719 TI - Poly(ADP-ribose) polymerase gene disruption renders mice resistant to cerebral ischemia. AB - Nitric oxide (NO) and peroxynitrite, formed from NO and superoxide anion, have been implicated as mediators of neuronal damage following focal ischemia, but their molecular targets have not been defined. One candidate pathway is DNA damage leading to activation of the nuclear enzyme, poly(ADP-ribose) polymerase (PARP), which catalyzes attachment of ADP ribose units from NAD to nuclear proteins following DNA damage. Excessive activation of PARP can deplete NAD and ATP, which is consumed in regeneration of NAD, leading to cell death by energy depletion. We show that genetic disruption of PARP provides profound protection against glutamate-NO-mediated ischemic insults in vitro and major decreases in infarct volume after reversible middle cerebral artery occlusion. These results provide compelling evidence for a primary involvement of PARP activation in neuronal damage following focal ischemia and suggest that therapies designed towards inhibiting PARP may provide benefit in the treatment of cerebrovascular disease. PMID- 9334720 TI - GLUT4 heterozygous knockout mice develop muscle insulin resistance and diabetes. AB - GLUT4, the insulin-responsive glucose transporter, plays an important role in postprandial glucose disposal. Altered GLUT4 activity is suggested to be one of the factors responsible for decreased glucose uptake in muscle and adipose tissue in obesity and diabetes. To assess the effect of GLUT4 expression on whole-body glucose homeostasis, we disrupted the murine GLUT4 gene by homologous recombination. Male mice heterozygous for the mutation (GLUT4 +/-) exhibited a decrease in GLUT4 expression in adipose tissue and skeletal muscle. This decrease in GLUT4 expression did not result in obesity but led to increased serum glucose and insulin, reduced muscle glucose uptake, hypertension, and diabetic histopathologies in the heart and liver similar to those of humans with non insulin-dependent diabetes mellitus (NIDDM). The male GLUT4 +/- mice represent a good model for studying the development of NIDDM without the complications associated with obesity. PMID- 9334721 TI - Transient renewal of thymopoiesis in HIV-infected human thymic implants following antiviral therapy. AB - Stem cell gene therapy strategies for AIDS require that differentiation-inducing stromal elements of HIV-infected individuals remain functionally intact to support the maturation of exogenous progenitor cells into mature CD4+ cells. To investigate the feasibility of stem cell reconstitution strategies in AIDS, we used the SCID-hu mouse to examine the ability of HIV-infected CD4+ cell-depleted human thymic implants to support renewed thymopoiesis. Here we report that following treatment of these implants with antiretroviral drugs, new thymopoiesis is initiated. This suggests that antiviral therapies might allow de novo production of T lymphocytes and provides support for the concept of therapeutic strategies aimed at reconstitution of the peripheral CD4+ T-cell compartment. PMID- 9334722 TI - Inactivation of HIV-1 chemokine co-receptor CXCR-4 by a novel intrakine strategy. AB - CXC-chemokine receptor (CXCR)-4/fusin, a newly discovered co-receptor for T-cell line (T)-tropic HIV-1 virus, plays a critical role in T-tropic virus fusion and entry into permissive cells. The occurrence of T-tropic HIV viruses is associated with CD4-positive cell decline and progression to AIDS, suggesting that the T tropic HIV-1 contributes to AIDS pathogenesis. In this study, we used a novel strategy to inactivate CXCR-4 by targeting a modified CXC-chemokine to the endoplasmic reticulum (ER) to block the surface expression of newly synthesized CXCR-4. The genetically modified lymphocytes expressing this intracellular chemokine, termed "intrakine", are immune to T-tropic virus infection and appear to retain normal biological features. Thus, this genetic intrakine strategy is uniquely targeted at the conserved cellular receptor for the prevention of HIV-1 entry and may be developed into an effective treatment for HIV-1 infection and AIDS. PMID- 9334723 TI - HIV-1 Vpr suppresses immune activation and apoptosis through regulation of nuclear factor kappa B. AB - The HIV-1 accessory gene product Vpr can influence viral pathogenesis by affecting viral replication as well as host cell transcription and proliferation. We have investigated the effects of Vpr on host cell activation and confirm that it influences cellular proliferation. However, we have also found that Vpr modulates T-cell receptor (TCR)-triggered apoptosis in a manner similar to that of glucocorticoids. In the absence of TCR-mediated activation, Vpr induces apoptosis whereas in its presence, Vpr interrupts the expected induction of apoptosis. This regulation of apoptosis is linked to Vpr suppression of NF-kappa B activity via the induction of I kappa B, an inhibitor of NF-kappa B. Further, Vpr suppresses expression of IL-2, IL-10, IL-12, TNF alpha and IL-4, all of which are NF-kappa B-dependent. The effects of Vpr could be reversed by RU486. Our finding that Vpr can regulate NF-kappa B supports the hypothesis that some aspects of viral pathogenesis are the consequence of cell dysregulation by Vpr. PMID- 9334724 TI - Interleukin-15 protects from lethal apoptosis in vivo. AB - Interleukin-15 shares many biological activities with IL-2 and signals through the IL-2 receptor beta and gamma chains. However, IL-15 and IL-2 differ in their controls of expression and secretion, their range of target cells and their functional activities. These dissimilarities may include differential effects on apoptosis. For example, IL-2 induces or inhibits T-cell apoptosis in vitro, depending on T-cell activation, whereas IL-15 inhibits cytokine deprivation induced apoptosis in activated T cells. Studying whether and how IL-15 modulates distinct apoptosis pathways, we show here that apoptosis induced by anti-Fas, anti-CD3, dexamethasone, and/or anti-IgM in activated human T and B cells in vitro is inhibited by IL-15 in a manner dependent on RNA synthesis. In vivo, anti Fas-induced lethal multisystem apoptosis in mice is suppressed by a novel IL-15 IgG2b fusion protein. Only IL-15, but not IL-2, completely protected from lethal hepatic failure. Thus, IL-15 is a potent, general inhibitor of apoptosis in vitro and in vivo with intriguing therapeutic potential. PMID- 9334725 TI - Testis-derived Sertoli cells have a trophic effect on dopamine neurons and alleviate hemiparkinsonism in rats. AB - Neural tissue transplantation has become an alternative treatment for Parkinson's disease (PD) and other neurodegenerative disorders. The clinical use of neural grafts as a source of dopamine for Parkinson's disease patients, although beneficial, is associated with logistical and ethical issues. Thus, alternative graft sources have been explored including polymer-encapsulated cells and nonneural cells (that is, adrenal chromaffin cells) or genetically modified cells that secrete dopamine and/or trophic factors. Although progress has been made, no current alternative graft source has ideal characteristics for transplantation. Emerging evidence suggests the importance of trophic factors in enhancing survival and regeneration of intrinsic dopaminergic neurons. It would be desirable to transplant cells that are readily available, immunologically accepted by the central nervous system and capable of producing dopamine and/or trophic factors. Sertoli cells have been shown to secrete CD-95 ligand and regulatory proteins, as well as trophic, tropic, and immunosuppressive factors that provide the testis, in part, with its "immunoprivileged" status. The present study demonstrated that transplantation of rat testis-derived Sertoli cells into adult rat brains ameliorated behavioral deficits in rats with 6-hydroxydopamine induced hemiparkinsonism. This was associated with enhanced tyrosine hydroxylase (TH) immunoreactivity in the striatum in the area around the transplanted Sertoli cells. Furthermore, in vitro experiments demonstrated enhanced dopaminergic neuronal survival and outgrowth when embryonic neurons were cultured with medium in which rat Sertoli cells had been grown. Transplantation of Sertoli cells may provide a useful alternative treatment for PD and other neurodegenerative disorders. PMID- 9334726 TI - Persistent infection with Theiler's virus leads to CNS autoimmunity via epitope spreading. AB - Multiple sclerosis (MS) is a T cell-mediated autoimmune demyelinating disease, which may be initiated by a virus infection. Theiler's murine encephalomyelitis virus (TMEV), a natural mouse pathogen, is a picornavirus that induces a chronic, CD4+ T cell-mediated demyelinating disease with a clinical course and histopathology similar to that of chronic progressive MS (ref. 3). Demyelination in TMEV-infected mice is initiated by a mononuclear inflammatory response mediated by virus-specific CD4+ T cells targeting virus, which chronically persists in the CNS (ref. 4-6). We show that beginning 3-4 weeks after disease onset, T-cell responses to multiple myelin autoepitopes arise in an ordered progression and may play a pathologic role in chronic disease. Kinetic and functional studies show that T-cell responses to the immunodominant myelin proteolipid protein epitope (PLP139-151) did not arise because of cross reactivity between TMEV and self epitopes (that is, molecular mimicry), but because of de novo priming of self-reactive T cells to sequestered autoantigens released secondary to virus-specific T cell-mediated demyelination (that is, epitope spreading). Epitope spreading is an important alternate mechanism to explain the etiology of virus-induced organ-specific autoimmune diseases. PMID- 9334727 TI - A secreted FGF-binding protein can serve as the angiogenic switch in human cancer. AB - The growth and metastatic spread of cancer is directly related to tumor angiogenesis, and the driving factors need to be understood to exploit this process therapeutically. However, tumor cells and their normal stroma express a multitude of candidate angiogenic factors, and very few specific inhibitors have been generated to assess which of these gene products are only innocent bystanders and which contribute significantly to tumor angiogenesis and metastasis. Here we investigated whether the expression in tumors of a secreted fibroblast growth factor (FGF)-binding protein (FGF-BP) that mobilizes and activates locally stored FGFs (ref. 11) can serve as an angiogenic switch molecule. Developmental expression of the retinoid-regulated FGF-BP gene is prominent in the skin and intestine during the perinatal phase and is down modulated in the adult. The gene is, however, upregulated in carcinogen-induced skin tumors, in squamous cell carcinoma (SCC) and in some colon cancer cell lines and tumor samples. To assess the significance of FGF-BP expression in tumors, we depleted human SCC (ME-180) and colon carcinoma (LS174T) cell lines of their endogenous FGF-BP by targeting with specific ribozymes. We found that the reduction of FGF-BP reduced the release of biologically active basic FGF (bFGF) from cells in culture. Furthermore, the growth and angiogenesis of xenograft tumors in mice was decreased in parallel with the reduction of FGF-BP. This suggests that human tumors can utilize FGF-BP as an angiogenic switch molecule. PMID- 9334728 TI - Growth hormone treatment induces mammary gland hyperplasia in aging primates. AB - The decline of growth hormone (GH) and insulin-like growth factor I (IGF-I) production during aging has been likened to the decrease in gonadal steroids in menopause. The repletion of GH/IGF-I levels in aging individuals is suggested to restore the lean tissue anabolism characteristic of youth. In addition to anabolic effects on musculo-skeletal tissues, GH also stimulates mammary glandular growth in some species, although its effects on primate mammary growth remain unclear. Some clinical observations implicate GH in human mammary growth, for example, gynecomastia occurs in some children treated with GH (ref. 6), and tall stature and acromegaly are associated with an increased incidence of breast cancer. To investigate the effects of GH/IGF-I augmentation on mammary tissue in a model relevant to aging humans, we treated aged female rhesus monkeys with GH, IGF-I, GH + IGF-I or saline diluent for 7 weeks. IGF-I treatment was associated with a twofold increase, GH with a three- to fourfold increase, and GH + IGF-I with a four'-to fivefold increase in mammary glandular size and epithelial proliferation index. These mitogenic effects were directly correlated with circulating GH and IGF-I levels, suggesting that either GH or its downstream effector IGF-I stimulates primate mammary epithelial proliferation. PMID- 9334730 TI - Bimoclomol: a nontoxic, hydroxylamine derivative with stress protein-inducing activity and cytoprotective effects. AB - Preservation of the chemical architecture of a cell or of an organism under changing and perhaps stressful conditions is termed homeostasis. An integral feature of homeostasis is the rapid expression of genes whose products are specifically dedicated to protect cellular functions against stress. One of the best known mechanisms protecting cells from various stresses is the heat-shock response which results in the induction of the synthesis of heat-shock proteins (HSPs or stress proteins). A large body of information supports that stress proteins--many of them molecular chaperones--are crucial for the maintenance of cell integrity during normal growth as well as during pathophysiological conditions, and thus can be considered "homeostatic proteins." Recently emphasis is being placed on the potential use of these proteins in preventing and/or treating diseases. Therefore, it would be of great therapeutic benefit to discover compounds that are clinically safe yet able to induce the accumulation of HSPs in patients with chronic disorders such as diabetes mellitus, heart disease or kidney failure. Here we show that a novel cytoprotective hydroxylamine derivative, [2-hydroxy-3-(1-piperidinyl) propoxy]-3-pyridinecarboximidoil chloride maleate, Bimoclomol, facilitates the formation of chaperone molecules in eukaryotic cells by inducing or amplifying expression of heat-shock genes. The cytoprotective effects observed under several experimental conditions, including a murine model of ischemia and wound healing in the diabetic rat, are likely mediated by the coordinate expression of all major HSPs. This nontoxic drug, which is under Phase II clinical trials, has enormous potential therapeutic applications. PMID- 9334729 TI - Tumor-selective transgene expression in vivo mediated by an E2F-responsive adenoviral vector. AB - Recent data suggest that many tumors, such as malignant gliomas, have disrupted pRB function, either because of RB-1 gene mutations or as a result of mutations affecting upstream regulators of pRB such as cyclin D1 or p16/INK4a/MTS1 (ref. 1 5). Tumor suppression by pRB has been linked to its ability to repress E2F responsive promoters such as the E2F-1 promoter. Thus, a prediction, which has not yet been demonstrated experimentally in vivo, is that E2F-responsive promoters should be more active in tumor cells relative to normal cells because of an excess of "free" E2F and loss of pRB/E2F repressor complexes. We demonstrate that adenoviral vectors that contain transgenes driven by the E2F-1 promoter can mediate tumor-selective gene expression in vivo, allowing for eradication of established gliomas with significantly less normal tissue toxicity than seen with standard adenoviral vectors. Our data indicate that de-repression of the E2F-1 promoter occurs in cancer cells in vivo, a finding that can be exploited to design viral vectors that mediate tumor-selective gene expression. PMID- 9334731 TI - Biallelic mutations in the ATM gene in T-prolymphocytic leukemia. AB - Ataxia-telangiectasia (AT) is an autosomal recessive disorder characterized by cerebellar ataxia, oculocutaneous telangiectasia, immune deficiency, genome instability and predisposition to malignancies, particularly T-cell neoplasms. The responsible gene, designated ataxia-telangiectasia mutated (ATM), was recently identified by positional cloning in the chromosomal region 11q22.3-23.1 (ref. 4, 5) ATM is 150 kb in length, consists of 66 exons and encodes a nuclear phosphoprotein of approximately 350 kDa (ref. 4-9). Although ATM is considered to be a tumorigenic factor in several human cancers, it has not yet been found mutated in tumors of non-AT patients. Given the marked predisposition of AT patients to develop neoplasms of the T-cell lineage, we analyzed a series of T cell leukemias (T-prolymphocytic leukemia, or T-PLL) in non-AT patients in search of genomic changes associated with the development of this disease. Among the recurrent aberrations identified, deletion of the chromosome arm 11q was very frequent. Subsequent molecular cytogenetic analyses allowed us to define a small commonly deleted segment at 11q22.3-23.1 in 15 of 24 T-PLLs studied. Since this critical region contained ATM, we further analyzed the remaining copy of the gene in six cases showing deletions affecting one ATM allele. In all six cases, mutations of the second ATM allele were identified, leading to the absence, premature truncation or alteration of the ATM gene product. Thus, our study demonstrates disruption of both ATM alleles by deletion or point mutation in T PLL, suggesting that ATM functions as a tumor-suppressor gene in tumors of non-AT individuals. PMID- 9334732 TI - The role of CCR5 and CCR2 polymorphisms in HIV-1 transmission and disease progression. AB - Entry of human immunodeficiency virus type 1 (HIV-1) into target cells requires both CD4 (ref. 1, 2) and one of a growing number of G-protein-coupled seven transmembrane receptors. Viruses predominantly use one, or occasionally both, of the major co-receptors CCR5 or CXCR4, although other receptors, including CCR2B and CCR3, function as minor co-receptors. CCR3 appears critical in central nervous system infection. A 32-base pair inactivating deletion in CCR5 (delta 32) common to Northern European populations has been associated with reduced, but not absolute, HIV-1 transmission risk and delayed disease progression. A more commonly distributed transition causing a valine to isoleucine switch in transmembrane domain I of CCR2B (64I) with unknown functional consequences was recently shown to delay disease progression but not reduce infection risk. Although we confirm the lack of association of CCR2B 64I with transmission, we cannot confirm the association with delayed progression. Although subjects with CCR5 delta 32 defects had significantly reduced median viral load at study entry, providing a plausible explanation for the association with delayed progression, this association was not seen with CCR2B 64I. Further studies are needed to define the role of CCR2B64I in HIV pathogenesis. PMID- 9334733 TI - Quantification of tumor cell invasion using confocal laser scan microscopy. PMID- 9334734 TI - A never ending frontier. PMID- 9334735 TI - A snapshot of a dynamic nuclear building block. PMID- 9334736 TI - Structural aspects of protein synthesis. PMID- 9334737 TI - Picture story. Pumping ions. PMID- 9334738 TI - Structure of a conserved RNA component of the peptidyl transferase centre. AB - The structure of a conserved hairpin loop involved in peptidyl-tRNA recognition by 50S ribosomal subunits has been solved by NMR. The loop is closed by a novel G C base pair and presents guanine residues for RNA recognition. PMID- 9334739 TI - Structure of benzyl T-antigen disaccharide bound to Amaranthus caudatus agglutinin. AB - Amaranthus caudatus agglutinin contains a novel arrangement of four beta-trefoil domains. The sugar-binding site provides specificity for the carcinoma-associated T-antigen disaccharide even when 'masked' by other sugars. PMID- 9334740 TI - The crystal structures of two spermadhesins reveal the CUB domain fold. AB - Spermadhesins, 12,000-14,000 M(r) mammalian proteins, include lectins involved in sperm-egg binding and display a single CUB domain architecture. We report the crystal structures of porcine seminal plasma PSP-I/PSP-II, a heterodimer of two glycosylated spermadhesins, and bovine aSFP at 2.4 A and 1.9 A resolution respectively. PMID- 9334742 TI - Saposin fold revealed by the NMR structure of NK-lysin. AB - NK-lysin is the first representative of a family of sequence related proteins- saposins, surfactant-associated protein B, pore forming amoeba proteins, and domains of acid sphingomyelinase, acyloxyacylhydrolase and plant aspartic proteinases--for which a structure has been determined. PMID- 9334741 TI - Structure of the receptor binding fragment HC of tetanus neurotoxin. AB - The 2.7 A structure of the tetanus neurotoxin receptor binding fragment Hc reveals a jelly-roll domain and a beta-trefoil domain. Hc retains the unique transport properties of the holotoxin and is capable of eliciting a protective immunological response against the full length holotoxin. PMID- 9334743 TI - Protein kinase inhibition by staurosporine revealed in details of the molecular interaction with CDK2. AB - Staurosporine exhibits nanomolar IC50 values against a wide range of protein kinases. The structure of a CDK2 staurosporine complex explains the tight binding of this inhibitor, and suggests features to be exploited in the design of specific inhibitors of CDKs. PMID- 9334744 TI - Structure of a kinetic protein folding intermediate by equilibrium amide exchange. AB - A combination of equilibrium amide exchange and kinetic folding data show that the essential features of the complex topology of the N-terminal domain of a thermophilic phosphoglycerate kinase are established on a millisecond or faster timescale, before the rate-limiting step in the folding pathway commences. PMID- 9334745 TI - Functional rapidly folding proteins from simplified amino acid sequences. AB - Early protein synthesis is thought to have involved a reduced amino acid alphabet. What is the minimum number of amino acids that would have been needed to encode complex protein folds similar to those found in nature today? Here we show that a small beta-sheet protein, the SH3 domain, can be largely encoded by a five letter amino acid alphabet but not by a three letter alphabet. Furthermore, despite the dramatic changes in sequence, the folding rates of the reduced alphabet proteins are very close to that of the naturally occurring SH3 domain. This finding suggests that despite the vast size of the search space, the rapid folding of biological sequences to their native states is not the result of extensive evolutionary optimization. Instead, the results support the idea that the interactions which stabilize the native state induce a funnel shape to the free energy landscape sufficient to guide the folding polypeptide chain to the proper structure. PMID- 9334746 TI - Structure and mechanism of endo/exocellulase E4 from Thermomonospora fusca. AB - Cellulase E4 from Thermomonospora fusca is unusual in that it has characteristics of both exo- and endo-cellulases. Here we report the crystal structure of a 68K M(r) fragment of E4 (E4-68) at 1.9 A resolution. E4-68 contains both a family 9 catalytic domain, exhibiting an (alpha/alpha)6 barrel fold, and a family III cellulose binding domain, having an antiparallel beta-sandwich fold. While neither of these folds is novel, E4-68 provides the first cellulase structure having interacting catalytic and cellulose binding domains. The complexes of E4 68 with cellopentaose, cellotriose and cellobiose reveal conformational changes associated with ligand binding and allow us to propose a catalytic mechanism for family 9 enzymes. We also provide evidence that E4 has two novel characteristics: first it combines exo- and endo-activities and second, when it functions as an exo-cellulase, it cleaves off cellotetraose units. PMID- 9334747 TI - Solution structure of the DNA-binding domain and model for the complex of multifunctional hexameric arginine repressor with DNA. AB - The structure of the monomeric DNA-binding domain of the Escherichia coli arginine repressor, ArgR, determined by NMR spectroscopy, shows structural homology to the winged helix-turn-helix (wHTH) family, a motif found in a diverse class of proteins including both gene regulators and gene organizers from prokaryotes and eukaryotes. Biochemical data on DNA binding by intact ArgR are used as constraints to position the domain on its DNA target and to derive a model for the hexamer-DNA complex using the known structure of the L-arginine binding domain. The structural independence of the wHTH fold may be important for multimeric DNA-binding proteins that contact extended DNA regions with imperfect match to consensus sequences, a feature of many wHTH-domain proteins. PMID- 9334748 TI - Crystal structure of nitric oxide reductase from denitrifying fungus Fusarium oxysporum. AB - Structures of nitric oxide reductase (NOR) in the ferric resting and the ferrous CO states have been solved at 2.0 A resolution. These structures provide significant new insights into how NO is reduced in biological systems. The haem distal pocket is open to solvent, implicating this region as a possible NADH binding site. In combination with mutagenesis results, a hydrogen-bonding network from the water molecule adjacent to the iron ligand to the protein surface of the distal pocket through the hydroxyl group of Ser 286 and the carboxyl group of Asp 393 can be assigned to a pathway for proton delivery during the NO reduction reaction. PMID- 9334749 TI - Structure of the collagen-binding domain from a Staphylococcus aureus adhesin. AB - The crystal structure of the recombinant 19,000 M(r) binding domain from the Staphylococcus aureus collagen adhesin has been determined at 2 A resolution. The domain fold is a jelly-roll, composed of two antiparallel beta-sheets and two short alpha-helices. Triple-helical collagen model probes were used in a systematic docking search to identify the collagen-binding site. A groove on beta sheet I exhibited the best surface complementarity to the collagen probes. This site partially overlaps with the peptide sequence previously shown to be critical for collagen binding. Recombinant proteins containing single amino acid mutations designed to disrupt the surface of the putative binding site exhibited significantly lower affinities for collagen. Here we present a structural perspective for the mode of collagen binding by a bacterial surface protein. PMID- 9334756 TI - A global effort to modernize otolaryngology-head and neck surgery: the Vietnam initiative. PMID- 9334757 TI - Cochlear implantation in children under the age of two years: candidacy considerations. AB - Within recent years, there has been a growing trend to lower the age at implantation below 2 years in children. The motivation for this trend is to provide children with access to auditory stimulation as early as possible, thereby taking advantage of crucial periods for speech and language development. Determining implant candidacy in very young children, however, poses numerous challenges and requires the development and evaluation of assessment procedures appropriate for this population. Issues in determining audiologic candidacy and evaluating implant benefit in young children are discussed. Measures that can be used to assess auditory development in this population and the application of some of these procedures in the pediatric study of the Clarion cochlear implant (Advanced Bionics Corp., Sylmar, Calif.) are also presented. PMID- 9334758 TI - Comparing implants with hearing aids in profoundly deaf children. AB - Speech perception, speech production, and spoken language skills of 13 age matched groups of prelingually deaf children, all enrolled in the same educational setting, were compared. Each group consisted of three children, two with hearing aids but different degrees of hearing loss (pure-tone average > 100 dB HL and pure-tone average between 90 and 100 dB HL) and one who had used a Nucleus multichannel implant (Cochlear Corp., Englewood, Colo.) for 3 years. The implanted group exhibited significantly better performance in all areas than aided children with greater than 100 dB losses. However, children with hearing losses in the 90 to 100 dB range who received hearing aids and auditory oral instruction at an early age performed just as well as children who had used implants for 3 years. PMID- 9334759 TI - Children with implants can speak, but can they communicate? AB - English-language skills were evaluated in two groups of profoundly hearing impaired children with the Reynell Developmental Language Scales, Revised. The first group consisted of 89 deaf children who had not received cochlear implants. The second group consisted of 23 children wearing Nucleus multichannel cochlear implants. The subjects without implants provided cross-sectional language data used to estimate the amount of language gains expected on the basis of maturation. The Reynell data from the group without implants were subjected to a regression by age. On the basis of this analysis, deaf children were predicted to make half or less of the language gains of their peers with normal hearing. Predicted language scores were then generated for the subjects with implants by using the children's preimplant Reynell Developmental Language Scale scores. The predicted scores were then compared with actual scores achieved by the subjects with implants 6 and 12 months after implantation. Twelve months after implantation, the subjects demonstrated gains in receptive and expressive language skills that exceeded by 7 months the predictions made on the basis of maturation alone. Moreover, the average language-development rate of the subjects with implants in the first year of device use was equivalent to that of children with normal hearing. These effects were observed for children with implants using both the oral and total-communication methods. PMID- 9334761 TI - Speech-processing strategies designed for children. AB - The design of cochlear prosthetic hardware and speech-processing strategies has been driven largely by psychophysical data from postlingually deafened adults with implants. In such subjects, success is related to the ability of the electrical stimulation to evoke the same patterns of neural activity and hence the same percepts as were produced formerly by acoustic input. Adults differ greatly in their ability to make use of information provided through electrical stimulation, particularly as temporal patterns. Recent research suggests that the manner in which information is processed in the auditory nervous system can be influenced by the type of information that is available during development of hearing. Because cochlear prostheses are used increasingly in prelingually deaf children, we must face the difficult task of designing and testing speech processing strategies that are more appropriate for developing nervous systems whose first and only experience with sound comes from electrical stimulation. PMID- 9334760 TI - Cognitive evoked potentials to speech and tonal stimuli in children with implants. AB - We investigated late and cognitive (mismatch negativity, P300) auditory potentials in 14 children with cochlear implants between the ages of 4 and 12 years. Length of cochlear implant use ranged from 7 to 84 months. Three types of stimulus contrasts were used: (1) a loudness contrast consisting of a 1500 Hz tone burst presented at 75 (standard) and 90 dB sound pressure level (deviant); (2) a frequency contrast consisting of a 1500 Hz tone burst (standard) and a 3000 Hz tone burst (deviant) presented at 80 dB sound pressure level; and (3) a speech contrast consisting of "heed" (standard) and "who'd" (deviant) delivered with a roving loudness paradigm involving a randomized variation of the levels of the standard and deviant stimuli. Latencies and amplitudes of components N1, P2, N2, and P3 and a mismatch negativity were measured. Overall, there were very few missing or unidentifiable components. P3 and mismatch negativity components were identified for all subjects and all stimuli. The latencies of most components were affected by stimulus type. There was a trend for longer latencies for the speech contrast compared with the loudness or frequency contrasts. This may be a reflection of the increased processing time required for the speech stimuli because of its higher complexity. There were several significant correlations between speech recognition and cognitive evoked potential latencies. These results indicate that the clinical use of cognitive evoked potentials in children with cochlear implants is feasible and informative. PMID- 9334762 TI - A model of educational resource use by children with cochlear implants. AB - We have tracked patterns of use of educational and rehabilitative resources as part of an initial assessment of cost-benefit ratios of cochlear implants in children. Forty-two children with cochlear implants in the Listening Center at Johns Hopkins program of aural rehabilitation have served as our study cohort to develop the measures to be assessed. An educational resource matrix stratifies school setting (residential vs special education vs non-specialized "mainstream" setting) and levels of rehabilitative support (speech and language therapy and interpreter use) to map past and current use of these services. Initial cost benefit projections based on observed advancement toward educational independence in the educational resource matrix indicate an extremely favorable net present value of the implant (cost savings minus cost). These cost-benefit projections will need to be supplemented with measures of the impact on quality of life and future educational and vocational options to determine the overall cost effectiveness of cochlear implants in children. PMID- 9334763 TI - Speech perception by prelingually deaf children using cochlear implants. AB - In this investigation we measured the performance of 50 prelingually deaf children on several speech perception tests. Children were from 2 to 15 years of age, and some children were tested with as much as 5 years of cochlear implant use. Speech perception tests included the recognition of stress pattern, consonants, vowels, words, and sentences. The audiovisual perception of consonants was also measured. Average results indicated that gains were being made in the perception of stress and words in a closed-set context within 1 year from implantation. The perception of words in an open-set context demonstrated much slower increases over time. Large individual differences were observed. Some preliminary data suggest that children who receive implants before the age of 4 years obtain higher scores, on average, than children who receive implants after the age of 5 years. Some children become part-time users or nonusers of their cochlear implants. The average results from 18 congenitally deaf children were significantly higher than the average results from 12 children with prelingually acquired deafness after 3 years of implant use. Information on vowel and consonant features shows increases in performance after 2 years of cochlear implant use, with the exception of the place feature. For this feature, no changes were observed. Vision-alone testing indicated that lipreading performance increased over time. An audiovisual enhancement provided by the cochlear implant was observed for all features. PMID- 9334764 TI - Parameter selection to optimize speech recognition with the Nucleus implant. AB - Speech coding strategy, frequency boundary assignment table, and speech processor program minimum and maximum stimulation levels are parameters of the Nucleus Cochlear Implant System whose selection affects speech recognition performance in adults and children. Research studies show that speech recognition is significantly better with (1) the Spectral Peak than with the Multipeak speech coding strategy and (2) frequency boundary assignment Table 7 than with Table 9 in an individual's speech processor program (MAP). Minimum and maximum stimulation levels in this MAP are based on psychophysical measurements on each electrode but often need to be modified for optimum use in everyday life. Many children and adults have increases, decreases, or fluctuations in electrical hearing that require changes in the MAP minimum and maximum levels to maintain their ability to recognize speech and other sounds. PMID- 9334765 TI - Observational assessments of the interaction of implant recipients with family and peers: preliminary findings. AB - Two experiments were conducted to determine the feasibility of direct observational methods in the assessment of social interactions between cochlear implant recipients and their families and peers. With laboratory analogs of parent-child interactions and peer interactions, children with implants and deaf children were videotaped. These videotapes were then coded for specific patterns of behavior. The results established the feasibility of direct observational assessments of deaf children and implant recipients and identified behavioral foci that could serve as effective indexes of implant outcome. The study also suggested that the peer entry paradigm may not be suitable for deaf children under the age of 7 years. PMID- 9334766 TI - Timing and trials of hearing aids and assistive devices. AB - Of all the factors under consideration during the evaluation for a cochlear implant, perhaps the most critical is the speech perception ability of the candidate when using appropriate amplification. For children, this issue is often complicated by inadequate training or limited use of the sensory aid. Guidelines for implantation suggest that there be a "lack of benefit from hearing aids" to qualify as a candidate. This "lack of benefit" must be explored when using the most appropriate aid coupled with the proper rehabilitation. In determining candidacy for children, it becomes extremely important to assess benefit using a variety of sensory aids while having access to well-directed auditory intervention. The types of amplification that can be used during the preimplant evaluation stage include conventional behind-the-ear devices, FM units, vibrotactile aids, and frequency transposition aids. Although some of these devices are used more often than others, centers implanting children should be aware of the role that each plays in the evaluation process. To determine the frequency with which these devices are used as well as the types of strategies that are emphasized in the preimplant training interval, a questionnaire was developed to gather information from the cochlear implant facilities in the United States. The results of this survey will be presented to demonstrate training and device trends that are incorporated during the preimplant process. Additionally, the use of frequency transposition hearing aids as part of the preimplant training procedure will be explored. The facility at Manhattan Eye, Ear and Throat Hospital has studied these devices in a small group of adults and children. Data pertaining to the speech perceptual abilities obtained with both the TranSonic and the Emily will be presented and compared to results with the Nucleus cochlear implant system. PMID- 9334767 TI - Cochlear implant soft surgery: fact or fantasy? AB - A basic surgical principle is to be as gentle as possible to accomplish the goals of the operation. The concept of "soft surgery" for cochlear implants consisted of a small, localized cochleostomy and gentle electrode insertion, the hope being that by limiting damage to the inner ear, superior hearing results might be obtained. The technique includes deferring the cochleostomy until immediately before electrode insertion, use of a large burr to flatten the promontory, followed by a smaller burr to expose the endosteum, preservation of the endosteum of the scala tympani, smoothing of the bony edges with burrs and dissectors, limited opening of the scala tympani, no suctioning of perilymph, gentle electrode insertion, and potential use of a lubricant to facilitate insertion. Although this technique has a theoretic basis, is esthetically satisfying, and has been used in many cases involving the Nucleus device at multiple centers, no data are available that demonstrate its advantages. Furthermore, the Clarion device, the results of which seem to be comparable to those of the Nucleus device, requires much more extensive and potentially damaging surgery. The pros and cons of soft surgery will be discussed. Although soft surgery seems desirable to limit trauma within the cochlea, other factors such as full electrode insertion, stimulation strategy, and survival of ganglion cells may be more important predictors of successful results. PMID- 9334768 TI - Cochlear implant in the child under two years of age: skull growth, otitis media, and selection. AB - Early cochlear implantation is desirable to effect maximal development of receptive and expressive language. Factors to consider in implanting before the age of 2 years include anatomic considerations, temporal bone growth and device extrusion or migration, the sequela of otitis media, and the accuracy of early diagnosis. Clinical experience and laboratory data indicate that cochlear implantation before the age of 2 years is possible. PMID- 9334769 TI - Patterns of neural degeneration in the human cochlea and auditory nerve: implications for cochlear implantation. AB - Although the identity of all the variables that may influence speech recognition after cochlear implantation is unknown, the degree of preservation of spiral ganglion cells is generally considered to be of primary importance. A series of experiments in our laboratories, directed at quantification of surviving spiral ganglion cells in the profoundly deaf, evaluation of the predictive value of a variety of clinical parameters, and the evaluation of the consequences of implantation in the inner ear, is summarized. Histologic study of the inner ears of patients who were deafened during life demonstrated that the cause of deafness accounted for 57% of the variability of spiral ganglion cell counts. Spiral ganglion cell counts were highest in individuals deafened by aminoglycoside toxicity or sudden idiopathic deafness and lowest in those deafened by postnatal viral labyrinthitis, congenital or genetic deafness, or bacterial meningitis. Study of the determinants of degeneration of the spiral ganglion revealed that degeneration is most severe in the basal compared with the apical turn and more severe when both inner and outer hair cells are absent. Unlike the findings in some experimental animal studies, no survival advantage of type II ganglion cells could be identified. There was a strong negative correlation between the degree of bony occlusion of the cochlea and the normality of the spiral ganglion cell count. However, even in specimens in which there was severe bony occlusion, significant numbers of spiral ganglion cells survived. A strong positive correlation between the diameter of the cochlear, vestibular, and eighth cranial nerves with the total spiral ganglion cell count (p < 0.001) was found. This would suggest that modern imaging techniques may be used to predict residual spiral ganglion cell population in cochlear implant candidates. Trauma from implantation of the electrode array was studied in both cadaveric human temporal bone models and temporal bones from individuals who received implants during life. A characteristic pattern of damage to the lateral cochlear wall and basilar membrane was identified in the upper basal turn. New bone formation and perielectrode fibrosis was common after cochlear implantation. Despite this significant trauma and reaction, there is no firm evidence that further degeneration of the spiral ganglion can be predicted as a consequence. PMID- 9334770 TI - Cochlear nucleus cell size changes in the dalmatian: model of congenital deafness. AB - We assessed cellular changes in one population of neurons of the cochlear nucleus associated with a form of genetic deafness in deaf dalmatians. Spheric cells from deaf dalmatians and age-matched control (hearing) dogs were analyzed morphometrically. The somatic silhouette of these cells was reduced by 22.1% to 38.1%. The effect on cell size was greater with increased duration of deafness. Because the deaf dalmatian exhibits progressive degeneration of the auditory periphery, shrinkage of spheric cells may reflect the initial influence of attenuated activity of auditory nerve fibers, and sensorineural degeneration with longer periods of deafness. PMID- 9334771 TI - Speech perception abilities of adult and pediatric Nucleus implant recipients using the Spectral Peak (SPEAK) coding strategy. AB - A series of 73 postlinguistically deafened adults and 34 prelinguistically deafened children were evaluated with the Spectral Peak (SPEAK) coding strategy of the Nucleus 22-channel cochlear implant. The adults who received consecutive implants demonstrated rapid acquisition of open-set speech recognition skills in the initial postoperative period. Group mean sentence recognition improved to 53.5% (n = 52) after 2 weeks, 62.1% (n = 55) after 1 month, 69.8% (n = 57) after 3 months, and 74.4% (n = 42) after 6 months of use. At the 6-month evaluation interval, 43% of subjects scored greater than 90% on sound-alone sentence recognition in quiet and only one patient (2.4%) scored less than 10%. Mean monosyllabic word recognition was 35.6% after 6 months of use. The 34 prelinguistically deafened children were converted from the Multipeak strategy to Spectral Peak strategy at four large pediatric implant centers. After 6 months of using the new coding strategy, the children demonstrated significant improvements in their speech perception abilities. PMID- 9334773 TI - Surgical techniques for cochlear implantation in the very young child. AB - Early cochlear implantation to treat prelingually deafened children has been shown to improve speech perception and overall performance. The current age limit for implantation is 24 months in accordance with US Food and Drug Administration guidelines, but it is believed that earlier implantation is possible and may result in better performance. Implantation in children younger than 36 months, however, is complicated by the altered anatomy of the temporal bone in this young age group. We have developed specific modifications in the cochlear implantation technique for this young age group. This technique was used in implantation for 17 children younger than 36 months. The ages ranged from 16 to 36 months and averaged 30 months. All patients except one had complete electrode insertion without complication. The technique of cochlear implantation must be modified not only for differences in anatomy in these young children but also for the expected continued growth of the temporal bone and related structures. Cochlear implantation can be safely performed on children as young as 16 months. PMID- 9334772 TI - Cochlear implants in children: what constitutes a complication? AB - The surgeon must ultimately accept the responsibility for any complications that occur as the result of a cochlear implant. Listening to the cochlear implant team members and responding to their needs may enhance the child's progress. Surgical complications, (i.e., skinflap problems, infection, and facial paralysis) are indeed infrequent, but nonsurgical problems are not. Surgical and nonsurgical experiences were reviewed in 55 children. Ages ranged from 23 months to 18 years at the time of cochlear implantation, which occurred from 1984 to 1995. There were no surgical complications. However, the most common surgical obstacle was ossification, which was present in 40% and undetected by computed tomographic scanning in 16.3% of children. Ossification occurred at the round window and scala tympani in 32.7% and involved the cochlea more extensively in 7.3% of children. In only one child was the cochlea entirely ossified. There were, however, many nonsurgical problems that were viewed as complications in patient management. The single most important complication was device failure. This occurred in 10.9% (5/46) of children with the Cochlear Corporation multichannel implant. Head banging and other temper tantrums, parental interference with rehabilitation, socioeconomic factors, poor compliance by the family unit, equipment problems, educational deficiencies, and impatience with habilitative training were some of the other problems. PMID- 9334774 TI - Electrical middle ear muscle reflex: use in cochlear implant programming. AB - Programming of multichannel cochlear implants (CIs) requires subjective responses to a series of sophisticated psychophysical percepts. It is often difficult for young prelinguistically deaf children to provide adequate responses for device fitting. This is especially true in setting levels of maximum comfortable loudness, whereby failure to indicate growth of loudness may result in elevation of stimulus levels to the threshold of pain. The acoustic or stapedial muscle reflex has been used previously to provide objective confirmation of acoustic stimulation, and there have been attempts to use the reflex in hearing aid fitting. It has also been suggested that electrically elicited middle ear muscle reflexes (eMEMR) may have applicability in confirming and quantifying electrical stimulation through a CI. To assess the relationship between eMEMR characteristics and levels of loudness perception with CIs, determine reliability of the response, and investigate potential use of eMEMR in CI programming, 25 postlinguistically deafened adult CI users were evaluated. Reflexes have also been attempted on 40 children, with responses present in 31 (71%). Comfort levels predicted by eMEMR were highly correlated with those obtained through subjective judgments in the adult subjects. The eMEMR provides an objective, accurate, and rapid method of estimating maximum comfortable loudness levels, which may be useful in the initial programming of young implant recipients. PMID- 9334775 TI - Increased secretory capacity of the middle ear mucosa after acute otitis media caused by Haemophilus influenzae type B. AB - Secretory otitis media is associated with a highly increased goblet cell density of the middle ear mucosa. Previous studies have shown that a single episode of experimental acute otitis media caused by Streptococcus pneumoniae or nontypeable Haemophilus influenzae is followed by increased goblet cell density for a period of at least 6 months. This condition may create a predisposition for subsequent development of secretory otitis media. We inoculated the middle ears of 25 rats with type B H. influenzae to determine the effect of the bacteria on mucosal secretory capacity. Five rats were euthanized 4, 8, 16, 60, and 180 days after inoculation, followed by dissection, staining, and whole-mount embedding of the middle ear mucosa. The goblet cell density was determined in 24 well-defined localities. Compared with that of 25 normal middle ears, the goblet cell density was significantly increased in almost all counting localities on all days of euthanasia. Thus increased goblet cell density and enlargement of mucosal areas containing goblet cells persisted for 6 months after the acute incident. Inoculation of type B H. influenzae induced an increase of goblet cell density that was higher than the increase after inoculation of S. pneumoniae or nontypeable H. influenzae. We conclude that experimental acute otitis media caused by type B H. influenzae is followed by a longstanding increase of mucosal secretory capacity, which is likely to induce a subsequent development of secretory otitis media. PMID- 9334777 TI - CO2 laser repair of permanent tracheostomy stricture. AB - We present our experience in treating postlaryngectomy patients with respiratory disturbance caused by stricture of the permanent stoma by CO2 laser surgery. Laser surgery is a simple procedure that can be performed with the patient under local anesthesia without bleeding and with minimal damage to the adjacent strictures resulting in a minimal postoperative edema and contracture. In our opinion the CO2 laser is a useful surgical tool for enlargement of the airway lumen and for improvement of respiratory disturbance in postlaryngectomy patients with stomal stricture, and this procedure should be considered for treatment of selected patients. PMID- 9334776 TI - Squamous cell carcinoma of the oral cavity in patients younger than 40 years. AB - Squamous cell carcinoma of the head and neck has been regarded as a disease affecting the elderly. Several etiologic factors have been demonstrated, such as tobacco and alcohol use and premalignant lesions, whereas others have been suspected, such as genetic or immunodeficiency disorders. Recently, some reports have addressed a tendency toward an increase in the incidence of squamous cell carcinoma in young patients. In recent years we have observed an increase in the number of squamous cell carcinomas in patients younger than 40 years. Therefore we retrospectively reviewed our clinical experience of cancer in those patients younger than 40 years. After screening 505 clinical charts, 294 patients met the criteria to enter our study. Twenty-four (8.2%) patients were aged 40 years or younger. Data collected included the history of premalignant lesions, etiologic factors, TNM stages, treatment modalities, and histopathologic issues. Statistical analysis with Kaplan-Meier survival rates and log-rank tests between various variables were applied. A significant association in survival was observed between patterns of recurrence (p = 0.031) and presence of neoplastic cells 5 mm or closer to the specimen margin. On the other hand, a lack of association was assessed in carcinogenic-related habits and in premalignant lesions. Likewise, although men showed a slightly worse prognosis than women, statistically no significant differences were found (p = 0.27). PMID- 9334778 TI - Horizontovertical laryngectomy for supraglottic carcinoma. AB - Recent decades have seen the expansion of indications for partial laryngectomy. This study focused on the application of horizontovertical laryngectomy for T2 and selected T3 and T4 supraglottic carcinomas. An osteomuscular flap was designed to reconstruct the laryngeal defect to preserve all the essential laryngeal functions. Seventy-six patients treated between November 1979 and October 1990 were reviewed retrospectively. All but two were followed up for at least 5 years. There were 11 cases of T2, 57 cases of T3, and 8 cases of T4 tumors. The survival rates of 7 cases of stage II, 56 cases of stage III, and 13 cases of stage IV carcinoma were 100%, 84.3%, and 74.3%, respectively. The functional recovery was satisfactory; 78.4% of 74 patients underwent decannulation, and all but one patient resumed a normal diet. This study showed that conservation surgery is reliable for selected supraglottic cancers, even for those lesions that extend beyond the confines of the supraglottis. PMID- 9334779 TI - Massive primary hemorrhage during tonsillectomy from a large venous varicosity. AB - The internal jugular vein is derived embryologically from the anterior cardinal veins. Developmental abnormalities may result in a cavernous venous varicosity, which may be the cause of massive primary hemorrhage during tonsillectomy. An index of suspicion is required for diagnosis, which may be confirmed by retrograde venography techniques. PMID- 9334780 TI - Geographic tongue. PMID- 9334781 TI - Pulsatile tinnitus illustrated. PMID- 9334782 TI - Adult rhinosinusitis defined. PMID- 9334783 TI - Clinical evaluation of rhinosinusitis: history and physical examination. PMID- 9334784 TI - Laboratory diagnosis. PMID- 9334785 TI - Rhinosinusitis: radiologic diagnosis. PMID- 9334787 TI - The medical management of rhinosinusitis. AB - The management of rhinosinusitis depends on a number of variables related to the duration and severity of symptoms in the individual patient. Furthermore acute rhinosinusitis is managed differently than chronic rhinosinusitis. Because a variety of conservative and pharmacologic interventions are available, the physician can find it difficult to develop a cohesive and logical approach to treatment. An understanding of the pathophysiology, microbiology, and natural history of rhinosinusitis is necessary to formulate the best treatment plan for the individual patient. PMID- 9334788 TI - Surgical management of adult rhinosinusitis. PMID- 9334786 TI - Staging for rhinosinusitis. AB - Interest in the surgical treatment of chronic rhinosinusitis has increased, primarily because rigid endoscopy and, more particularly, computed tomographic scanning have facilitated the visualization of disease. At the same time it has become both scientifically and financially imperative to audit therapeutic outcome. Consequently, a staging system for nonneoplastic sinus disease is needed. It is clear that any assessment of medical or surgical therapeutic response requires a method of quantifying disease severity that will be widely accepted by practitioners in the field. This acceptance will largely depend on how easy the method is to apply. With computed tomographic scanning it is possible to more accurately determine the extent of the pathologic condition in rhinosinusitis, a disease in which the severity of symptoms and the appearances on nasal endoscopy have a significantly more unpredictable correlation with the extent of disease. One goal of the Task Force on Rhinosinusitis of the American Academy of Otolaryngology-Head and Neck Surgery was to recommend a system for outcomes research that combines quantification with ease of application. PMID- 9334789 TI - Pediatric rhinosinusitis. PMID- 9334790 TI - Outcomes assessment. PMID- 9334791 TI - Direct reimbursement: implications for research. PMID- 9334792 TI - An educational intervention as decision support for menopausal women. AB - The purpose of this study was to develop and test a decision support intervention (DSI) to assist women to make and act on informed decisions that are consistent with their values in the area of menopause and hormone replacement therapy (HRT). Mode and intensity of intervention were tested in midlife women (N = 248), randomly assigned to one of three intervention formats: written information only, guided discussion, or personalized decision exercise. Data were collected over 12 months. Knowledge, decisional conflict, satisfaction with health care provider, and self-efficacy improved following intervention and were maintained for 12 months for all groups. Women's adherence to their own plans over 12 months was 59% (exercise), 76% (calcium intake), and 89% (HRT). Carefully written information is effective in promoting knowledge, adherence, and satisfaction among well-educated, interested women. It was concluded that women can understand complex information, including tradeoffs regarding treatment options. Women will adhere to their own plans, suggesting that consumer rather than provider plans may be the more appropriate gold standard for measuring adherence. PMID- 9334793 TI - Experiences of fatigue and depression before and after low-dose 1-thyroxine supplementation in essentially euthyroid individuals. AB - Hypothyroidism is a relative state, long associated with fatigue and depression. Individuals may experience thyroid-related symptoms such as fatigue and depression before thyroid indices become abnormal. However, because of clinicians' diverse interpretations of appropriate circumstances for its use, low dose, 1-thyroxine supplementation often is overlooked as a therapeutic agent for symptom treatment. The purpose of this exploratory, hermeneutic study was to describe euthyroid individuals' experiences of fatigue and depression before and after low-dose 1-thyroxine supplementation. For women participants, the collective influence of fatigue and depression prior to treatment interfered significantly with their day-to-day lives, despite their euthyroid status. For men, the influence of symptoms was far less substantial than for women. In general, participants responded favorably, both physically and emotionally, to low-dose 1-thyroxine supplementation. Furthermore, no participant experienced 1 thyroxine induced hyperthyroidism or untoward side effects attributable to 1 thyroxine. Further study of effects of 1-thyroxine on symptoms is needed. PMID- 9334794 TI - Early family life and victimization in the lives of women. AB - The purpose of this study was to test a causal model designed to identify relationships among early family life experiences, including abuse; cognitive coping mechanisms and social support; and victimization in adult women. Women (N = 622) from several communities in Northeast Ohio responded to public announcements of the study and completed a questionnaire comprised of 10 scales measuring the model variables. The theoretical structures of the original model did not fit the empirical data. Nonsignificant paths were deleted and nonestimated paths that maximally improved fit were added sequentially to reconstruct the model. The reconstructed model included a network of multiple, mediated pathways leading to the outcome variable, current victimization. Although childhood abuse experiences and victimization throughout adulthood predicted current victimization, the general emotional health of the family of origin predicted victimization beyond the effects accounted for by earlier abuse experiences. The cognitive variables mediated the relationships between early and later abuse. PMID- 9334795 TI - Weight loss methods used by African American and Euro-American women. AB - In this naturalistic study employing intensive interviews and anthropometric measures, an educationally and economically diverse community-based sample of 40 African American and 40 Euro-American women described their lifetime experiences with weight management. Twenty types of weight loss methods were identified and grouped into one of three categories: lifestyle work, head work, and professional services. The most frequently used weight loss methods were from the lifestyle work category, with the leading methods identified as exercise on own and reduce high calorie and/or increase low calorie foods. African Americans and Euro Americans overwhelmingly used similar weight loss methods, with the only significant difference occurring in the more frequent use of commercial diet products among the African American group. Methods from the head work category were used significantly more often by women with higher social status, while heavier women more frequently sought professional services to lose weight than thinner women. The Euro-American women engaged in weight loss methods for significantly longer periods of time and were found to weigh significantly less than the African American women. These findings suggest that the shorter duration of weight loss attempts may be a major factor contributing to the larger body size in African American women. PMID- 9334796 TI - Short-term regulation of energy intake is intact in hypophagic tumor-bearing rats. AB - Anorexia and weight loss are major problems in the care of cancer patients. Data from laboratory studies using an animal model of tumor-induced anorexia suggest that energy intake may be regulated in the tumor-bearing host as it is in healthy animals. The purpose of the present study was to determine if hypophagic tumor bearing rats would alter their food intake in response to manipulations known to affect food intake in normal healthy animals. Both tumor-bearing and healthy animals increased their food intake when housed at 22 versus 25 degrees C and reduced their food intake when injected with anorexigenic doses of bacterial lipopolysaccharide or interleukin-1 alpha. These data suggest that selected physiological responses affecting short-term food intake are intact in the hypophagic tumor bearing host. PMID- 9334798 TI - Nurses' job satisfaction, absenteeism, and turnover after implementing a special care unit practice model. AB - The purpose of the study was to compare job satisfaction, absenteeism, and turnover between nurses working in a nurse-managed special care unit (SCU) and those working in traditional intensive care units (ICU). A case management practice model with a shared governance management model and minimal technology was implemented in the SCU while contrasting features of a primary nursing practice model with a bureaucratic management model and high technology already in place in the traditional ICU. Individual nurses' perceptions of and their preferences for the SCU practice model also were examined related to job satisfaction. Using analysis of covariance, greater satisfaction with a lower absenteeism rate was found in nurses working in the SCU. Nurses' perceptions and preferences for the SCU practice model were closely related to their job satisfaction and growth satisfaction. The findings suggest that individual perception and preference should be taken into account before implementing autonomy, authority, and responsibility at the organizational level to lead to the desired nurse outcomes in a given working environment. PMID- 9334797 TI - Psychophysiological response to a laboratory challenge in women with and without diagnosed irritable bowel syndrome. AB - The purpose of this study was to describe and compare physiological variables at baseline and in response to laboratory stress among women diagnosed with irritable bowel syndrome (IBS, n = 26), women with undiagnosed chronic gastrointestinal symptoms consistent with IBS (IBS-NP, n = 24), and asymptomatic women (n = 22). Urine catecholamine levels were measured in the first voided specimen on the morning of testing. Cardiovascular variables were measured at baseline and repeatedly during the Stroop Color-Word Conflict Test (Stroop). Women in the IBS group had higher baseline systolic blood pressure (SBP) than the control group and higher basal urine norepinephrine (NE) levels than the IBS-NP group. Control for activity or body mass reduced the group difference in SBP to nonsignificance but did not affect the observed difference in urine NE. There were no significant differences among the groups in other baseline values or in response to the Stroop. These results suggest that, despite higher basal urine catecholamine levels, cardiovascular reactivity to a cognitive challenge in a laboratory setting is not elevated in women with diagnosed IBS. PMID- 9334799 TI - Occupational and nonoccupational factors in job satisfaction and psychological distress among nurses. AB - To facilitate nurses' job satisfaction and reduce their psychological distress, it is useful for a nursing manager to know whether factors within the workplace provide greater prediction of these affective states than variables outside the domain of work, and whether there are common predictors of satisfaction and distress. The relative importance of occupational and nonoccupational variables in the prediction of job satisfaction and psychological distress was investigated in a survey of hospital nurses (N = 376). Perceived relations with the head nurse, coworkers, physicians, and other units/departments, along with unit tenure and job/nonjob conflict, were predictors of job satisfaction. Personal disposition (anxiety-trait), social integration, unit tenure, professional experience, position level, and job/nonjob conflict, along with the relations with the head nurse and physicians, were predictors of psychological distress. The relations with the head nurse and physicians, as well as unit tenure and job/nonjob conflict, were predictors of both satisfaction and distress. The prediction by unit tenure is noteworthy. Unit tenure had a negative relationship to satisfaction and a positive one to distress, whereas total experience had a negative relationship to psychological distress and none with job satisfaction. The role of unit tenure in nurses' affective experiences warrants more attention in future research, along with the role of job/nonjob conflict and other variables predictive of nurses' satisfaction and distress. PMID- 9334800 TI - Interpreting kappa values for two-observer nursing diagnosis data. AB - Reliability of nursing observations often is estimated using Cohen's kappa, a chance-adjusted measure of agreement between observer RNs. However, use of kappa as an omnibus measure sometimes can be misleading. In a study partly designed to describe the frequency and reliability of nursing diagnoses in long-term care facilities, 360 residents each were assessed independently by two registered nurses, and kappa and observed proportion of agreement were calculated as estimates of reliability. For some diagnoses we observed high proportions of agreement, yet paradoxically low kappa values. This article presents an in-depth statistical analysis to resolve this paradox. Results from our analysis also suggest means for planning improvements in the diagnostic performance of participating RNs. Consequently, our approach can be used in similar studies of diagnosis reliability to enhance nursing research, education, and practice. PMID- 9334801 TI - Effect of parity and age at delivery on breast cancer risk in Slovenian women aged 25-54 years. AB - In 1988, a case-control study on breast cancer and oral contraceptives with 624 cases and 624 matched controls in the age range 25-54 years was undertaken in Slovenia. This analysis assesses the relationship between parity and breast cancer risk: the relative importance of age at first birth, age at subsequent births and total parity. We also evaluate whether a dual effect of an increased risk immediately after childbirth followed by a long-term benefit exists. Three logistic regression models were used. Age at first delivery is an important breast cancer risk factor: among parous women it was associated with a 5.3% increase/year in the odds of breast cancer. Multiparity was not shown to be an independent risk factor. Age at subsequent deliveries was associated with a 1% increase in risk for every 1 year increase of age at any birth, but this contribution to the risk was not significant. In the analysis stratified by parity the most important influence is with the age at first birth. We find no evidence of an effect on the odds of breast cancer associated with the age at the second, or later, births. We do find that there is an increased risk associated with the birth of the first child followed by a longer term protective effect. A post-menopausal woman has a reduced breast cancer risk compared with a pre menopausal woman of the same age, adjusting for the same number of deliveries and ages at these deliveries. PMID- 9334802 TI - Expression of fibronectin and tenascin-C mRNA by myofibroblasts, vascular cells and epithelial cells in human colon adenomas and carcinomas. AB - To understand the mechanisms of tissue remodeling during cancer progression, it is important to know the type of cells that actively express extracellular matrix (ECM) proteins. Twenty-nine adenocarcinomas, 5 adenomas and non-neoplastic mucosa samples were therefore investigated to determine their fibronectin (FN) and tenascin-C (TN-C) expression using in situ hybridization (ISH) and immunohistochemical staining. In the non-neoplastic mucosa, no mRNA signals were found. Two of the adenomas demonstrated positive signals in peri-cryptal cells and the vessels. In the cancers, TN-C and FN mRNAs were found in 86% and 96% of the total cases, respectively. The signals were mainly detected in myofibroblasts, labeled with alpha-smooth muscle actin, in the cancer stroma. TN C mRNA-positive cells were often observed in localized areas, such as in cancer stroma associated with invading edges and/or in host tissues surrounding the invading cancer front, but rarely in the center of the tumors. FN mRNA-positive cells were more widely spread throughout the cancer stroma, although they were also frequently observed at invading edges. Vascular cells in cancer tissues were also labeled. In 10 specimens, cancer cells themselves expressed FN and/or TN-C mRNA. Comparison with histo-pathological findings revealed positive relationships between the degree of mRNA expression of FN and TN-C and the depth of invasion as well as the frequency of metastasis to lymph nodes. The expression of FN and TN-C by myofibroblasts, vascular cells and cancer cells could be important for the remodeling process of neoplastic tissues during cancer development and progression. PMID- 9334803 TI - Serum levels of soluble tumor-necrosis-factor receptors in patients with benign and malignant HPV-associated anogenital lesions. AB - The levels of type-I and type-II soluble TNF-alpha receptors (sTNF-Rs) were evaluated in sera from patients with various human-papillomavirus-(HPV) associated benign and malignant anogenital lesions using specific enzyme-linked immunobiological assays. In patients with benign HPV6/11-associated condylomata acuminata, the levels of sTNF-RI were significantly increased, while sTNF-RII were in normal range. Both types of sTNF-Rs were in normal range in patients with benign HPV16-associated grade-I/II and grade-III cervical intra-epithelial neoplasia. However, their levels were significantly increased in patients with HPV16/18-associated squamous cervical cancer and anogenital Bowen's carcinoma. Sera from patients with condylomata acuminata and anogenital carcinomas displayed significantly increased TNF-alpha-inhibitory activity, as revealed by L929 cell cytotoxicity assay. Increased serum TNF-alpha-inhibitory activity correlated with higher levels of sTNF-Rs. Furthermore, this inhibitory activity could be specifically abrogated by htr9 and utr1 monoclonal antibodies recognizing TNF-RI and TNF-RII respectively. Our results strongly suggest that serum sTNF-Rs may protect tumor cells from cytotoxic/cytostatic effects of locally released TNF alpha, and that elevated levels of circulating sTNF-Rs may facilitate the growth of HPV-associated anogenital lesions. PMID- 9334804 TI - DNA repair proficiency in breast cancer patients and their first-degree relatives. AB - Defective DNA repair capacity as measured by enumerating chromatid aberrations induced in G2 phase by X-irradiation may explain increased risk of breast cancer among relatives of patients. In the present study, chromatid damage was determined in peripheral blood lymphocytes (PBL) following in vitro exposure to 50R X-irradiation in G2 phase from 14 breast cancer (BrCa) patients, 19 first degree relatives (FDR) of BrCa patients and 17 control women who had no family history of cancer for the last 3 generations. Controls, BrCa patients and their FDR had comparable frequency of gaps and breaks when cells were arrested with Colcemid (30 min) after X-irradiation. A steep decline in chromatid damage was observed in cells of controls when arrested after 30, 90 and 120 min of X irradiation. BrCa patients and their FDR showed higher frequencies of lymphocytic chromatid damage as compared to controls. Chromatid damage (95 gaps + breaks per 100 cells) observed among controls at 90 min post X-irradiation was considered as the optimal level of efficient DNA repair. Thirty-five percent of controls, 93% of BrCa patients and 79% of FDR showed sub-optimal DNA repair. Amongst the FDR, the likelihood of having suboptimal DNA repair was 7 times higher and the risk of developing breast cancer was 2.7 times higher as compared to controls. Moreover, in the BrCa patients, there was frequent involvement of chromosomes 1 and 2, and chromosomes of B, D and E groups, while in FDR, involvement of chromosome 2 and chromosomes of B, D and E groups was more frequent. PMID- 9334805 TI - Photodynamic therapy using mTHPC for malignant disease in the oral cavity. AB - Photodynamic therapy (PDT) produces local tumor necrosis, on activation of a previously administered sensitizer with non-thermal light of an appropriate wavelength. It is attractive for treating tumors of the mouth as tissue healing is particularly good. We describe the use of the photosensitizing agent meta tetrahydroxyphenyl chlorin (mTHPC, Foscan) for PDT of oral cancer, including patients with field cancerization. Nineteen patients with histologically confirmed oral cancer (8 with field change disease) and one with severe dysplasia, were sensitised with mTHPC intravenously. Activation was carried out 72-96 hr later with laser light at 652 nm using a range of light doses. The results were assessed clinically and histologically. Multiple biopsies were taken during the ulcerative stages to look at the effects of PDT and after healing to assess the overall treatment result. All single lesions up to stage T3 cleared after one PDT treatment (total of 6 patients). Three out of 6 T4 tumours were also cleared. Lesions in patients with field change disease did less well, only 9 of 14 T1 and T2s clearing, including 4 that required extra treatments with a higher light dose. Most healed very well, but tongue tethering was seen in 1 patient and another had necrosis in normal areas due to light scattering within the mouth. PDT using mTHPC is a promising new treatment for patients with oral cancer. PMID- 9334806 TI - Amplification of the mdm-2 gene and p53 abnormalities in uterine sarcomas. AB - The aim of this study is to address the role of mdm-2-gene amplification in the tumorigenesis of uterine sarcomas. Differential PCR with DNA from formalin-fixed paraffin-embedded specimens was employed in 12 patients with uterine sarcomas. We detected mdm-2-gene amplification in 4 out of 12 uterine sarcomas. The estimated copy number of the mdm-2 gene ranged from 4 to 13. Positive cases included 1 leiomyosarcoma and 3 carcinosarcomas, however, there was no correlation between mdm-2-gene amplification and clinicopathological characteristics. Over-expression of p53 protein was also immunohistochemically studied in the same series of patients: 4 out of 8 carcinosarcomas displayed p53 immunoreactivity. Taking these results together, only one carcinosarcoma was found to have both mdm-2-gene amplification and p53 over-expression. In contrast, half of the patients were found to have alterations either of mdm-2 or of p53. These findings support the notion that mdm-2-gene amplification might be an alternative mechanism for escaping from the regulatory pathway of p53 to suppress cell growth. PMID- 9334807 TI - Interaction between bcl-2 and p21 (WAF1/CIP1) in breast carcinomas with wild-type p53. AB - The bcl-2 protein is found to be over-expressed in many types of human tumours and is a potent inhibitor of apoptosis. The exact mechanism by which bcl-2 prevents apoptosis and exercises its oncogenic effect is still unclear. Other studies on cell lines have reported that bcl-2 over-expression is related to suppression of p21 (WAF1/CIP). We have investigated the relationship between bcl 2 protein over-expression and expression of the p21 protein in a series of human breast carcinomas. Selected tumour samples from 100 breast-cancer patients (38 with abnormal p53 status, scored as protein accumulation and/or mutation, and 62 without detectable p53 alterations), were immunostained for bcl-2 protein, the p21 protein and the oestrogen-receptor (ER) protein. A highly significant association was found between reduced p21-protein expression and over-expression of bcl-2 in tumours with no detectable p53 alterations (p < 0.001). A significant association was seen between ER immunoreactivity and expression of the bcl-2 protein, as well as between bcl-2 protein expression and tumours of the higher differentiation grade (grade-2 tumours). No association was seen between bcl-2 over-expression and the presence of metastases. Our findings indicate that down regulation of p21 may be a result of up-regulation of bcl-2 independent of p53. PMID- 9334808 TI - Selection of tumor antigens as targets for immune attack using immunohistochemistry: I. Focus on gangliosides. AB - Understanding the distribution of tumor-associated antigens on cancers and normal tissues is essential for selection of targets for cancer immunotherapy. Seven carbohydrate antigens, potential targets for immunotherapy, were studied using a panel of well-characterized MAbs by immunohistochemistry on cryostat-cut tissue sections of 13 types of cancers and 18 normal tissues. GD2 and GD3 were present on most cancers of neuroectodermal origin and GD2 was also present on B cell lymphomas. 9-O-acetyl-GD3 was detected only on melanoma while fucosyl GM1 was detected only on small cell lung cancers (SCLC). Surprisingly, GM2 was strongly expressed on all tested tumors, including cancers of neuroectodermal origin and cancers of epithelial origin. Polysialic acid was primarily expressed on SCLC and neuroblastomas. Globo H was present on most cancers of epithelial origin. These antigens were also identified in normal tissues. Fucosyl GM1 was not expressed significantly on any of the normal tissues analyzed. GD3, GD2, GM2 and polysialic acid were detected in normal brain to varying degrees. GM2 and Globo H were expressed on the luminal surface of epithelia of a variety of organs. The unexpected expression of GM2 on a broad range of cancers and normal epithelial tissues was confirmed by loss after methanol fixation and by immune thin layer chromatography. PMID- 9334809 TI - Selection of tumor antigens as targets for immune attack using immunohistochemistry: II. Blood group-related antigens. AB - Blood group-related antigens have been attractive targets for immunotherapy of cancer since their initial identification as cancer-related antigens. However, available information on the relative expression of most of these antigens on human malignant and normal tissues has been insufficient for selecting optimal antigens and tumors for immune attack. In this study, the distribution of the blood group-related antigens TF, Tn, sTn, Le(a), sialyl Le(a), Le(b), Le(x), sialyl Le(x), polyfucosyl Le(x) and Le(y) on 13 types of cancer and 16 normal tissues was compared. Our results show that sTn is strongly expressed on cancers of breast, colon, stomach, ovary, prostate and uterus; Tn on prostate cancer; TF on cancers of breast, colon, ovary, prostate and uterus; Le(y) on the cancers of colon, lung, pancreas and ovary; Le(a) and Le(x) on gastric cancer; and sialyl Le(a) and sialyl Le(x) on colon cancer. The complete absence of these antigens on cancers of neuroectodermal or mesodermal origin including melanoma, sarcoma, neuroblastoma and B cell lymphoma is as striking as their widespread presence on tumors of epithelial origin. Normal tissues were also tested. Tn and Le(b) were only detected on gastric and ovarian epithelia; sTn on Leydig cells of testis in addition to gastric and ovarian epithelia; Le(x) and sialyl Le(x) on polymorphonuclear leukocytes; and TF, Le(a), sialyl Le(a), Le(x), sialyl Le(x), polyfucosyl Le(x) and Le(y) on epithelia from a variety of tissues. PMID- 9334811 TI - Epidemiology of bladder cancer in Alexandria, Egypt: tobacco smoking. AB - The relationship between smoking and bladder cancer risk was investigated using data from a case-control study conducted between January 1994 and July 1996 in Alexandria, Egypt. Cases were 151 males with incident, histologically confirmed invasive cancer of the bladder, and controls were 157 males admitted to hospital for acute, non-neoplastic, non-urinary tract, non-smoking-related conditions. With reference to never smokers, ex-smokers had a multivariate odds ratio (OR) of 4.4 [95% confidence interval (CI) 1.7-11.7] and current smokers of 6.6 (95% CI 3.1-13.9). The ORs were 5.4 for < 20 and 7.6 for > or = 20 cigarettes per day. After adjustment for cigarette smoking, the ORs were 0.8 for waterpipe and 0.4 for hashish smokers. The risk was significantly related to duration of smoking (OR of 16.5 for > 40 years), and inversely related to age at starting (OR of 8.8 for starting < 20 years), and inversely related to time since quitting smoking. Compared with never smokers who did not report a clinical history of schistosomiasis, the OR was 9.4 for smokers with a history of schistosomiasis, and 10.7 for smokers ever employed in high-risk occupations compared with non smokers not reporting such a history. Thus, our results, while not giving indications of an increased bladder cancer risk with habits other than cigarette smoking, found a remarkably strong association with various measures of cigarette smoking that could explain 75% of bladder cancer cases among males from Alexandria. The prevalence of smoking was very low among women, and consequently tobacco was not a relevant risk factor for female bladder cancer. PMID- 9334810 TI - Hemizygous or homozygous deletion of the chromosomal region containing the p16INK4a gene is associated with amplification of the EGF receptor gene in glioblastomas. AB - The p16INK4a gene product acts as a negative regulator of the cell cycle by binding to cyclin-dependent kinases (CDKs) 4 and 6, thereby inhibiting the formation of an active CDK/cyclin D complex. Deletion of the p16 locus has been observed in tumor cell lines and, less frequently, in primary human neoplasms. We analyzed 31 glioblastomas and identified 6 cases with hemizygous and 6 with homozygous deletions of the p16 locus. Eight of these cases showed a concurrent amplification of the EGFR gene (epidermal growth factor receptor) while the overall frequency was 35%. This close correlation suggests that deletion of the p16 chromosomal region constitutes another genetic hallmark of the primary glioblastoma, which rapidly develops de novo, without a less malignant precursor lesion and for which EGFR amplification is a characteristic genetic change. The p16 protein was not detectable in 15 of 22 glioblastomas but only 4 of these showed homozygous deletion of the gene. The alternative transcript p16 beta, for which a growth-suppressing function has been suggested, was co-expressed with p16 alpha mRNA in most cases. Hypermethylation of CpG islands in the 5' region of the p16 gene was identified in only 1 case, suggesting that this alternative mechanism of gene silencing is rarely responsible for loss of p16 expression in glioblastomas. Likewise, only 1 glioblastoma carried a p16 mutation and in addition, unexpectedly, a homozygous deletion of p16 in approximately 80% of tumor cells. This mutation, Arg24Pro, has previously been identified in a melanoma kindred. PMID- 9334812 TI - Overexpression of autocrine TGF-beta 1 suppresses the growth of spindle epithelial cells in vitro and in vivo in the rat 4NQO model of oral carcinogenesis. AB - We examined the effect of the stable transfection of latent TGF-beta 1 cDNA, under the control of a cytomegalovirus promoter in the expression vector pcDNA3, into a 4NQO-induced clonal rat oral keratinocyte cell line that formed undifferentiated spindle cell tumours following subcutaneous transplantation to athymic mice. Test cells containing latent TGF-beta 1 cDNA produced a 2.3-fold increase in TGF-beta 1 protein compared to pcDNA3 controls as demonstrated by ELISA. Neutralisation experiments indicated that the majority of the protein was in the latent form. Untransfected and transfected (containing either TGF-beta 1 cDNA or pcDNA3) cell lines were keratin negative and vimentin positive. Cells transfected with TGF-beta 1 were inhibited more than pcDNA3 controls when cultured in an anchorage dependent or independent environment. Subcutaneous transplantation of cells overproducing TGF-beta 1 resulted in tumours of significantly smaller volume than vector-only controls. Further, orthotopic transplantation of cells containing TGF-beta 1 cDNA to the floor of the mouth in athymic mice markedly inhibited the development of pulmonary metastases compared to vector-only controls. Both test and control cell lines in athymic mice formed undifferentiated tumours with a complete absence of keratin elaboration. Subcutaneous xenografts were recultured and cells containing the TGF-beta 1 cDNA produced a similar amount of TGF-beta 1 peptide as the cells containing pcDNA3 only. The production of TGF-beta 1 by both of the xenograft-derived cell lines was significantly less than the parent, pre-transplanted cell lines and the untransfected cell line. All of the cell lines were inhibited by exogenous TGF beta 1. Our results demonstrate that autocrine TGF-beta 1 functions as a tumour suppressor in vitro and in vivo in 4NQO-induced spindle tumour cells that are growth inhibited by the ligand. Furthermore, tumour formation in athymic mice is associated with selection for a cell phenotype with diminished autocrine TGF-beta 1 production. PMID- 9334813 TI - Differential sensitivity of leukemic and normal hematopoietic progenitors to the killing effect of hyperthermia and quercetin used in combination: role of heat shock protein-70. AB - Autologous bone-marrow transplantation (ABMT) is widely used in the treatment of acute leukemias where a matched sibling donor is not available for allogeneic transplantation. However, a major problem in ABMT is relapse, and ex vivo purging may be very important in preventing it. We show here that quercetin enhances the growth-inhibitory effect of hyperthermia (HT) in AML (19 cases) and ALL (6 cases) leukemic blasts. Furthermore, the inhibitory effect of this combined treatment resulted in leukemic-cell apoptosis. On the contrary, normal hematopoietic progenitors were neither growth-inhibited nor induced to apoptosis by HT-plus quercetin treatment. To explain this difference in sensitivity of leukemic and normal hematopoietic progenitors, we analyzed the effect of quercetin on heat induced expression of heat-shock protein-70 (HSP-70), which has been shown to be important in regulating thermosensitivity. We found that quercetin inhibits heat induced HSP-70 expression both at protein and at mRNA levels in AML and ALL blasts. In normal CD34+ progenitors, the combined treatment with HT and quercetin did not reduce HSP-70 expression and did not induce cell apoptosis. Considering the difference in heat sensitivity of normal CD34+ and leukemic progenitors in the presence of quercetin, the combined use of HT and quercetin could constitute a purging protocol for ABMT. PMID- 9334814 TI - Expression of the multidrug resistance-associated protein (MRP) and chemoresistance of human non-small-cell lung cancer cells. AB - Human non-small-cell lung cancer (NSCLC) is considered to be a chemotherapy refractory malignancy. The underlying mechanisms remain rather obscure. The multidrug resistance-associated protein (MRP), mediating a multidrug resistance (MDR) phenotype, has been reported to be overexpressed in several drug-selected lung cancer cell lines. A few previous studies have described intrinsic MRP expression in both NSCLC and normal lung tissues. However, the drug-transporting activity as well as the correlation with chemoresistance is unclear. Using 15 unselected cell lines, we show that MRP (mRNA and protein as detected by reverse transcriptase polymerase chain reaction and immunoblot) is frequently expressed intrinsically, with markedly varying intensity, in NSCLC. Two cell lines expressed high MRP levels, one comparable to the drug-selected controls (GLC4/ADR, HL-60/AR) without, however, amplification of the MRP gene (Southern hybridization). Using 3H-daunomycin (3H-DM) and calcein as MRP substrates and probenecid (PRO), genistein (GEN), benzbromarone (BB), N-ethylmaleimide (NEM) and verapamil (VP) as MRP modulators, drug accumulation studies revealed a transporting activity of MRP that correlated significantly with the gene expression data. Moreover, a significant correlation between MRP expression and chemoresistance against daunomycin (DM), doxorubicin (DOX), etoposide (VP-16) and vinblastine (VBL), but not cisplatin (CDDP) and bleomycin (Bleo) (MTT-based survival assay), was detected. Correlations mainly rested on the pronounced chemoresistance of 2 highly MRP-expressing cell lines and did not reach significance when these cell lines were excluded. PMID- 9334815 TI - Enhanced tumor-forming capacity for immortalized melanocytes expressing melanoma growth stimulatory activity/growth-regulated cytokine beta and gamma proteins. AB - Three human MGSA/GRO genes encode 3 highly related chemokines, MGSA/GRO alpha, beta and -gamma. All 3 MGSA/GRO proteins bind to the same receptors, but with differing affinities, and stimulate a number of biological responses including chemotaxis, angiogenesis, and growth regulation. We have previously demonstrated that MGSA/GRO alpha can be isolated from culture medium conditioned by malignant melanoma cells and that continuous secretion of MGSA/GRO alpha contributes to the transformation of immortalized murine melanocytes. The present study was designed to determine whether MGSA/GRO beta or -gamma have similar effects on melanocyte tumorigenicity. Stable Melan-a clones expressing either human MGSA/GRO beta or gamma exhibited enhanced ability to form large colonies in soft agar and tumors in nude mice. The clones expressing the MGSA/GRO beta or -gamma transgene formed tumors within 2 months after injection; the tumors were highly pigmented and expressed immunoreactive MGSA/GRO beta or -gamma protein. Furthermore, when conditioned medium from Melan-a clones expressing MGSA/GRO alpha, -beta or -gamma transgenes were examined for the ability to induce angiogenesis in the rat cornea, strong angiogenic responses were observed. This angiogenic response was blocked by antibodies to the respective MGSA/GRO protein, but not by normal rabbit serum. By contrast, angiogenic responses were observed in only 2 of 12 corneal implants (17%) containing medium conditioned by Melan-a clones expressing the neomycin resistance marker alone. PMID- 9334816 TI - Characterization of the effects of the novel non-steroidal antiestrogen EM-800 on basal and estrogen-induced proliferation of T-47D, ZR-75-1 and MCF-7 human breast cancer cells in vitro. AB - Since estrogens play a predominant role in the development and growth of human breast cancer, antiestrogens represent a logical approach to the treatment of this disease. The present study compares the effects of the novel nonsteroidal anti-estrogen EM-800 and related compounds with those of a series of anti estrogens on basal and 17 beta-estradiol (E2)-induced cell proliferation in human breast cancer cell lines. In the absence of added E2, EM-800 and related compounds failed to change basal cell proliferation, thus showing the absence of intrinsic estrogenic activity in the ER-positive T-47D, ZR-75-1 and MCF-7 cell lines. The stimulation of T-47D cell proliferation induced by 0.1 nM E2 was competitively blocked by a simultaneous incubation with EM-652, EM-800, OH tamoxifen, OH-toremifene, ICI 182780, ICI 164384, droloxifene, tamoxifen and toremifene at apparent Ki values of 0.015, 0.011-0.017, 0.040-0.054, 0.043, 0.044, 0.243 and 0.735 nM, approx. 10 nM and > 10 nM, respectively. Similar data were obtained in ZR-75-1 and/or MCF-7 cells. Moreover, EM-652 was 6-fold more potent than OH-Tamoxifen in inhibiting the proportion of cycling MCF-7 cells. Our data show that EM-800 and EM-652 are the most potent known antiestrogens in human breast cancer cells in vitro and that they are devoid of the estrogenic activity of OH-tamoxifen and droloxifene suggested by stimulation of cell growth in the absence of estrogens in ZR-75-1 and MCF-7 cells. PMID- 9334817 TI - Ornithine decarboxylase inhibitor lessens the rat gastric carcinogenesis enhancement caused by tyrosine methyl ester. AB - The effects of combined administration of a catecholamine precursor, tyrosine methyl ester (TME), and an ornithine decarboxylase (ODC) inhibitor, 1,3 diaminopropane (DAP), on the incidence of gastric cancers induced by N-methyl-N' nitro-N-nitrosoguanidine (MNNG), the norepinephrine (NE) concentration and ODC activity of the gastric wall, and the labeling index of the gastric mucosa were investigated in inbred Wistar rats. Rats received s.c. injections of TME, 512 mg/kg body weight, every other day and drinking water with or without 2.5 g/l of DAP after 25 weeks of oral administration of MNNG. At week 52, administration of TME resulted in significant increases in the incidence of gastric cancers, in the NE concentration and the ODC activity of the antral portion of the gastric wall, and in the labeling index of antral epithelial cells. Administration of both TME and DAP significantly reduced the enhancements by TME of gastric carcinogenesis, NE concentration and ODC activity of the antral wall, and the labeling index of the antral mucosa. Our results suggest that ODC inhibition lessens enhancement by TME of gastric carcinogenesis and that the enhancement by TME of gastric carcinogenesis is mediated in part by polyamine biosynthesis. PMID- 9334818 TI - Targeted therapy of schwannoma cells in immunocompetent rats with an erbB2 specific antibody-toxin. AB - Over-expression of the erbB2-receptor tyrosine kinase is frequently observed in many human tumors of epithelial origin. Due to its causal involvement in malignant transformation and its presence on the tumor cell surface erbB2 is an attractive target for directed tumor therapy. We earlier described the potent anti-tumoral activity of the recombinant single-chain antibody toxin scFv(FRP5) ETA in vitro and in nude mouse tumor models in vivo. This molecule consists of the variable domains of the heavy and light chains of an erbB2-specific antibody genetically fused to a truncated Pseudomonas exotoxin A. Here we have investigated the in vivo effects of this immunotoxin on erbB2 expressing NV2Cd schwannoma cells growing as s.c. tumors in syngeneic BDIX rats. Established tumors were treated either locally by intra-tumoral injection of scFv(FRP5)-ETA or systemically by injection into the tail vein. Both routes of application resulted in pronounced inhibition of tumor growth with local treatment being more effective. Treatment with 25 micrograms/day of scFv(FRP5)-ETA for 10 days suppressed tumor growth almost completely. Antibodies directed mainly against the toxin domain of the fusion protein developed in all animals treated. PMID- 9334819 TI - Identification of HLA-A*0201-restricted CTL epitopes encoded by the tumor specific MAGE-2 gene product. AB - MAGE-2 is expressed in many tumors, including melanoma, laryngeal tumors, lung tumors and sarcomas, but not in healthy tissue, with the exception of testis. Thus, MAGE-2-derived peptides that bind to HLA class I molecules and elicit cytotoxic T lymphocyte (CTL) responses could be of significant therapeutic importance. In this study, we show that several MAGE-2-derived peptides bind with high affinity to HLA-A*0201. Three of them form complexes with HLA-A*0201 that are stable at 37 degrees C and are immunogenic in HLA-A*0201Kb transgenic mice. Moreover, CTLs against 2 of them (M2 112-120, and M2 157-166) specifically recognize cells that express both the MAGE-2 protein and HLA-A*0201Kb. These 2 peptides are processed and presented in the context of HLA-A*0201. Therefore, these peptides are candidate components in peptide-based vaccines for the treatment and prevention of several types of MAGE-2-expressing cancers. PMID- 9334820 TI - Role of the genetic background of rats in infection by HTLV-I and HTLV-II and in the development of associated diseases. AB - Three aspects of the rat model of HTLV-I/II infection were investigated. (i) The efficacy of HTLV-I-transformed rat cell lines in infecting different strains of rats: WKY and Lewis HTLV-I-transformed cell lines were injected into adult WKY, Lewis and Brown Norway rats, representing syngeneic and allogeneic combinations. The HTLV-I provirus was not detected in peripheral-blood mononuclear cells (PBMC) from these rats 18 weeks after inoculation, showing that HTLV-I-transformed rat cells are not suitable for virus challenge in vaccination experiments. Rats inoculated with Lewis HTLV-I-transformed cells produced an antibody response to HTLV-I, which was higher in allogeneic (WKY and Brown Norway) than in syngeneic rats. (ii) The susceptibility of rats to HTLV-II infection: After human HTLV-II producing cells (MO) were injected into adult WKY rats, the HTLV-II provirus was detected in PBMC 12 weeks later. Sequencing of a portion of this provirus confirmed its identity with the HTLV-II from MO cells. (iii) The role of MHC haplotype in susceptibility to neurological disease in rats inoculated as newborns with HTLV-I: The hypothesis that the RT-Ik haplotype confers susceptibility was tested by inoculating newborn OKA (RT-Ik), WKY (RT-Il), Lewis (RT-Il) and Fischer 344 (RT-I lvl) rats with human HTLV-I-producing cells (MT-2). Eighteen months later, only the WKY rats showed histological abnormality of the spinal cord, without clinical paralysis. Fischer 344 rats developed cutaneous tumors and OKA rats mammary tumors. The HTLV-I provirus was not detected in these tumors. PMID- 9334821 TI - Novel exon connections of the brain-specific (BS) exon of the adenomatous polyposis coli gene. AB - Expression of the adenomatous polyposis coli (APC) gene derived exon BS (e.g., brain-specific exon) has been analyzed by RT-PCR. Four novel APC mRNA isoforms derived from alternative splicing of exons 1A, BS and 1 were identified, which were ubiquitously expressed. One novel cDNA was characterized by cloning and DNA sequence analysis, which combined the exon 1A (identical with exon 0.3) 3' end with nucleotide position +101 of intron 1A and continued throughout exon BS. A second cDNA isoform was isolated, which joined the 3' end of exon 1A with nucleotide position +118 of exon BS. Both novel isoforms were found to be expressed together with a third novel APC exon connection, which was specified by exon BS/2 joining. This interesting exon junction resulted in novel deduced amino terminal open reading frames, which are completely in-frame with sequences located further downstream. Systematic exon connection analyses revealed that APC transcripts with exon BS/2 junctions were predominantly detected with a fixed exon composition. RT-PCR analyses did not identify facultative skipping of exons 9, 10A and 14 in this type of mRNA, in contrast to exon 1-containing APC transcripts analyzed from the same cDNA pool under identical conditions. Hence, exon 1 skipping of exon BS-positive mRNA molecules might preferentially encode unique APC polypeptide chains, which are characterized by an alternative amino terminus and extended heptad repeat structures due to combined incorporation of exons 9 and 10A. PMID- 9334822 TI - Generation and phenotypic characterization of new human ovarian cancer cell lines with the identification of antigens potentially recognizable by HLA-restricted cytotoxic T cells. AB - This study describes a simple method for long-term establishment of human ovarian tumor lines and prediction of T-cell epitopes that could be potentially useful in the generation of tumor-specific cytotoxic T lymphocytes (CTLs). Nine ovarian tumor lines (INT.Ov) were generated from solid primary or metastatic tumors as well as from ascitic fluid. Notably all lines expressed HLA class I, intercellular adhesion molecule-1 (ICAM-1), polymorphic epithelial mucin (PEM) and cytokeratin (CK), but not HLA class II, B7.1 (CD80) or BAGE. While of the 9 lines tested 4 (INT.Ov1, 2, 5 and 6) expressed the folate receptor (FR-alpha) and 6 (INT.Ov1, 2, 5, 6, 7 and 9) expressed the epidermal growth factor receptor (EGFR); MAGE-1 and p185HER-2/neu were only found in 2 lines (INT.Ov1 and 2) and GAGE-1 expression in 1 line (INT.Ov2). The identification of class I MHC ligands and T-cell epitopes within protein antigens was achieved by applying several theoretical methods including: 1) similarity or homology searches to MHCPEP; 2) BIMAS and 3) artificial neural network-based predictions of proteins MAGE, GAGE, EGFR, p185HER-2/neu and FR-alpha expressed in INT.Ov lines. Because of the high frequency of expression of some of these proteins in ovarian cancer and the ability to determine HLA binding peptides efficiently, it is expected that after appropriate screening, a large cohort of ovarian cancer patients may become candidates to receive peptide-based vaccines. PMID- 9334823 TI - Hepatocyte growth factor stimulates motility, chemotaxis and mitogenesis in ovarian carcinoma cells expressing high levels of c-met. AB - A proportion of ovarian carcinomas markedly overexpress the proto-oncogene c-met, which encodes the receptor for hepatocyte growth factor (HGF). HGF may either stimulate or inhibit the multiplication of its target cells, and may also promote motogenesis and morphogenesis. In this study, we established that the ovarian carcinoma-derived cell-line SK-OV-3 expressed about 20-fold higher levels of c met protein than are expressed by a second line, CH1. This enabled us to test functional consequences of high-level expression of c-met in ovarian carcinoma cells. The addition of HGF to attached cultures of SK-OV-3 cells caused a change to a motile phenotype, that was evident after 4-6 hr and affected essentially all of the cells by 24 hr. When HGF was placed in the lower compartment of a migration chamber, it induced a 17-fold increase in the migration of SK-OV-3 cells to the lower surface of the filter. Finally, HGF stimulated the incorporation of [3H]-thymidine by cultures of SK-OV-3 cells incubated in medium containing either low (0.2%) or full (10%) FCS. None of these responses were obtained when HGF was added to CH1 cells. We conclude that high levels of c-met expression in ovarian cancer cells may lead to a range of responses to HGF that would promote tumour growth and dissemination. PMID- 9334824 TI - Potent therapeutic activity of irinotecan (CPT-11) and its schedule dependency in medulloblastoma xenografts in nude mice. AB - The anti-tumor activity of irinotecan (CPT-11), a DNA-topoisomerase 1 inhibitor, was evaluated in 5 advanced stage subcutaneous medulloblastoma xenografts in nude mice, using different schedules of administration. With a 5-day schedule, the highest i.v. dose tested (40 mg kg-1 day-1) induced complete regressions in all xenografts but 1, and delays in tumor growth always exceeded 30 days. Two xenografts, IGRM11 and IGRM33, were highly sensitive, and animals survived tumor free beyond 120 days after treatment. CPT-11 clearly retained its anti-tumor activity at a lower dosage (27 mg kg-1 day-1). CPT-11 was significantly more active than cyclophosphamide, thiotepa and etoposide against the 3 xenografts evaluated. To study the schedule dependency of its anti-tumor activity, CPT-11 was given i.v. at the same total doses over the same period (33 days) using either a protracted or a sequential schedule in IGRM34-bearing mice. With a dose of 10 mg kg-1 day-1 given on days 0-4, days 7-11, days 21-25 and days 28-32 (total dose, 200 mg kg-1), 3 of 6 animals were tumor free on day 378. The same total dose given with a sequential schedule, i.e., 20 mg kg-1 day-1 on days 0-4 and days 28-32, failed to induce complete regression. The plasma pharmacokinetics of CPT-11 and SN-38 were studied in IGRM34-bearing animals after a single i.v. dose of 10 and 40 mg kg-1. The plasma clearance rate of CPT-11 was dose dependent. The ratio between the SN-38 and CPT-11 area under the curve in plasma was 0.4-0.65, i.e., significantly higher than that observed in humans at the maximum tolerated dose (0.01-0.05). Conversely, this ratio was 10-fold lower in tumor than in plasma. Clinical development of irinotecan is warranted in pediatric malignancies. PMID- 9334825 TI - Increased MRP expression is associated with resistance to radiation, anthracyclines and etoposide in cells treated with fractionated gamma-radiation. AB - The failure of chemotherapy is often associated with the failure of radiotherapy in the treatment of cancer. To investigate this relationship, the CCRF-CEM (CEM) human T-cell leukaemia cell line was treated with fractionated gamma-radiation totalling 75 Gy (10 cycles of 1.5 Gy daily for 5 days). This produced the CEMRR subline which was 1.5-fold resistant to radiation compared with the parental CEM cells. The CEMRR subline was also resistant to daunorbicin, idarubicin and etoposide but not to paclitaxel, cis-platinum or chlorambucil. Treatment with 50 microM buthionine sulphoximine, an inhibitor of glutathione synthesis, reversed the daunorubicin resistance in the CEMRR subline. Multidrug resistance-associated protein (MRP) mRNA was 6-fold higher in the CEMRR subline than in the CEM cells, and there was no detectable expression of P-glycoprotein in either the CEM cells or the CEMRR subline. Treatment of the CEM cells with 2 Gy of gamma-radiation caused an increase in MRP-mRNA within 4 hr which, by 24 hr, was greater than 5 fold that of the untreated CEM cells. No change in MRP mRNA was observed in the CEMRR subline with similar treatment. We conclude that MRP is involved in the immediate response to radiation and it may account for the drug resistance that often develops following radiation treatment. PMID- 9334826 TI - Experimental induction of liver fibrosis in young guinea pigs by combined application of copper sulphate and aflatoxin B1. AB - Aflatoxin B1 alone (0.05 mg resp. 0.037 mg/kg/d), copper alone (6.6 mg/kg/d or 200 mg/l drinking water) or a combination of both was administered orally for 6 months to young guinea pigs from the first/second day of life. In the copper group there were no pathomorphological changes. For the aflatoxin B1 group, liver damage was established. In the combined group, liver injury was more frequent and more severe compared to the aflatoxin B1 group and biliary copper excretion was diminished compared with the copper group. Histologically, only the livers of this group exhibited degeneration, atrophy and steatosis of liver cells, inflammatory processes and a more or less prominent fibrosis. For childhood cirrhosis (ICC and ICT) a combined etiology--a liver damaging agent plus elevated alimentary copper--is a plausible hypothesis. PMID- 9334827 TI - Lead levels in 24-h urine in Japanese adults. AB - Samples of 24-h urine were collected from 278 subjects (159 males and 119 females) in 1985, and 321 subjects (161 males and 160 females) in 1993 in the same factory with no occupational exposure to lead (Pb) in Japan. The age range of the subjects was 20-59 years. The urinary Pb concentrations were analyzed by flameless atomic absorption spectrophotometry after wet digestion followed by solvent extraction. The Pb levels in 24-h urine were distributed log-normally with geometric means (geometric S.D.) of 4.74 (1.46) and 2.67 (1.98) micrograms/day for males; and 3.22 (1.42) and 2.14 (2.05) micrograms/day for females in 1985 and 1993, respectively. No age-related changes in Pb levels in 24 h urine in either sex from 20 to 59 years were apparent. Pb levels in 24-h urine were significantly higher in males than in females (P < 0.01). Analysis of male smokers together with age-matched non-smokers failed to show an elevation of Pb levels in 24-h urine as related to smoking habits. Comparison of the Pb levels in 24-h urine in 1985 and 1993 disclosed that the Pb levels in 24-h urine were significantly lower in 1993 than in 1985 in both sexes (P < 0.01). PMID- 9334828 TI - Pharmacokinetics of tertiary butyl alcohol in male and female Fischer 344 rats. AB - Tertiary butyl alcohol (TBA) is a small aliphatic alcohol with multiple industrial and scientific uses. A comprehensive pharmacokinetic profile for TBA has not been determined in rats. The purpose of this study was to fully characterize the pharmacokinetics of TBA in male and female F-344 rats following intravenous administration of 37.5, 75, 150 and 300 mg/kg TBA. TBA was observed to undergo a rapid distribution phase followed by a slower elimination phase. The steady-state volume of distribution for TBA was roughly 4.5 times greater than total body water, and the clearance was lower than the estimated glomerular filtration rate. The elimination of TBA appears to saturate at higher doses, as evidenced by a disproportional increase in area under the concentration-time curve and decreased rate of clearance. PMID- 9334829 TI - Detection of trichloroethylene-protein adducts in rat liver and plasma. AB - Trichloroethylene is an industrial chemical with widespread occupational exposure and is a major environmental contaminant. In a Western blot using antiserum that recognizes trichloroethylene covalently bound to protein, a single 50 kDa microsomal adduct was detected in the livers of trichloroethylene-treated Sprague Dawley rats. To determine if trichloroethylene-protein adducts could be detected in blood, plasma proteins were immunoaffinity purified using an antidichloroacetyl column. A single 50 kDa protein was detected in the affinity purified fraction in a Western blot using dichloroacetyl antiserum. This protein was also immunochemically reactive with anti-cytochrome P450 2E1 antibodies. The 50 kDa trichloroethylene-protein adduct may be formed in the liver and released into the blood following exposure to trichloroethylene. The significance of adduct formation with respect to trichloroethylene toxicity remains to be established; however, the data suggest that this approach may be useful in the investigation of trichloroethylene-protein adducts and adverse effects following exposure. PMID- 9334830 TI - Evaluation of combined toxic effects of GB/GF and efficacy of jielin injection against combined poisoning in mice. AB - A computer program (Q-test) was used to evaluate the combined toxic effects of nerve agent GF and its combined form with sarin (GB/GF) in mice. Efficacy of Jielin Injection, the 2-PAM-containing antidote used successfully in China for the treatment of organophosphate pesticide poisoning, was also evaluated and compared with HI-6 against single and combined poisonings. The two agents were basically additive in toxicity when combined. However, toxic signs (convulsions) appeared later in combined poisoning than after exposure to each agent alone. The protective ratio of Jielin Injection against GF poisoning was low but significantly higher when against poisoning by GB or combined agent. When HI-6 was substituted for 2-PAM, the antidote was more effective against poisoning by both single and combined agents. Results of in vitro reactivation of GF-inhibited human erythrocyte acetylcholinesterase by these oximes agreed with the in vivo antidotal efficacy. PMID- 9334831 TI - Arsenic-induced changes in certain neurotransmitter levels and their recoveries following chelation in rat whole brain. AB - Arsenic as sodium arsenite (100 ppm in drinking water) was administered to male rats for 16 weeks. Animals were then treated either with meso-2,3 dimercaptosuccinic acid (DMSA), sodium 2,3-dimercaptopropane 1-sulfonate (DMPS), dimethyl DMSA (DmDMSA), or diisopropyl DMSA (DiPDMSA) twice daily (50 mg/kg) intraperitoneally for 5 days. After 5 days of rest period, the animals were again given a second course of chelation therapy. The animals were sacrificed subsequently for the determination of whole brain biogenic amines levels, acetylcholinesterase (AChE), monoamine oxidase (MAO) and delta-aminolevulinic acid dehydratase (ALAD) activities. A number of biochemical parameters and arsenic concentrations in some tissues were also determined. The results suggest a significant increase in brain arsenic concentration accompanied by alterations in neurotransmitters levels following As(III) exposure. Although chelation treatment was effective in reducing As burden, the altered biochemical variables responded less favorably to chelation therapy. The DMSA-diesters, particularly DiPDMSA, produced a more pronounced increase in brain arsenic burden, as well as alterations in a few neurotransmitters. It can be concluded that the lipophilic character of As antidotes may lead to unfavorable results following intraperitoneal administration. PMID- 9334833 TI - Developmental toxicity of 2-butin-1,4-diol following oral administration to the rat. AB - Developmental toxicity of 2-butin-1,4-diol was determined in groups of 18-22 pregnant Wistar rats at dose levels of 10, 40 and 80 mg/kg bw/day administered by gavage from days 6 to 15 pc. At 80 mg/kg bw/day food consumption and maternal body weight were reduced and one dam died during the treatment period. At this dose level the incidence of affected fetuses per litter with accessory 14th ribs was increased. This variation is assessed as an embryotoxic effect resulting from non-specific stress on the dams. No teratogenic effects were caused by 2-butin 1,4-diol. The NOAEL on the maternal and the developing organism was 40 mg/kg bw/day. PMID- 9334832 TI - Four-week repeated inhalation study of HCFC 225ca and HCFC 225cb in the common marmoset. AB - Four male and three female marmosets in each group were exposed to air only, 1000 ppm of HCFC 225ca or 5000 ppm of HCFC 225cb, for 6 h per day for 28 consecutive days. HCFC 225ca caused a slight reduction in body weight. HCFC 225cb occasionally caused somnolence during exposure and vomiting on the first day of exposure. Clinical chemistry findings included a mild reduction of triglyceride, cholesterol and phospholipid levels and increased GOT level in the HCFC 225ca exposure group. HCFC 225cb also caused a reduction of triglyceride levels in some animals. HCFC 225ca caused a slight increase of hepatic carnitine palmitoyltransferase (CPT) activity while HCFC 225cb slightly increased cyanide insensitive palmitoyl CoA beta-oxidation (FAOS) activity. In the HCFC 225cb exposure group, an increase in cytochrome P-450 content was also observed. HCFC 225ca caused a fatty change in the hepatic cells. Increased incidence of lipid droplets in the hepatic cells and myelin-like bodies in hepatic cells, Kupffer's cells and hepatic blood vessels were observed electron microscopically in the HCFC 225ca exposure group. A proliferation of smooth endoplasmic reticulum was observed in the HCFC 225cb exposure group. Decreased peroxisome volume density in the HCFC 225ca group, and increased volume density in the HCFC 225cb exposed females were seen. However, organ weight measurement and histopathological examination did not reveal hepatomegaly or hypertrophy with either substance. Although slight changes were noticed in peroxisome volume density and in some of the peroxisomal enzyme activities, the changes related to peroxisome proliferation with HCFC 225ca and 225cb were minimal in marmosets compared to those seen in rats. Histopathological examination and hormonal analysis did not reveal any abnormalities in the pancreas or testes. PMID- 9334834 TI - Lack of involvement of retinoic acid receptor alpha in retinoid-induced skin irritation in hairless mice. AB - It has been proposed that RAR gamma, the major retinoic acid receptor (RAR) subtype in skin, mediates retinoid-induced skin irritation. However, RAR alpha is also found in skin, and its role in retinoid-induced skin irritation has not been tested. In this study, RAR subtype-specific agonists and antagonists were used to test the possible contribution of RAR alpha to retinoid-induced skin irritation. Female hairless mice were treated topically on the dorsal skin for 5 days with various retinoids over a 2-log dose range, and cutaneous toxicity was scored by semiquantitative visual observations of skin flaking and abrasions daily up to 3 days post-treatment. Three RAR alpha-selective agonists were > or = 100-fold less potent as skin irritants than the structurally-related RAR pan-agonist, TTNPB. Skin irritation potency decreased in the following order: TTNPB > > Am580 > AGN 193835 > > 193836 and correlated with RAR beta and/or RAR gamma binding affinity rather than RAR alpha binding affinity. TTNPB-induced skin irritation was blocked in a dose-dependent fashion by co-treatment with the RAR pan-antagonist AGN 193109 but was not blocked by co-treatment with the RAR alpha-specific antagonist AGN 194301. In contrast, skin irritation induced by the RAR alpha-selective agonist AGN 193835 was almost completely blocked by co-treatment with AGN 193644, an RAR beta/gamma-selective antagonist. These data demonstrate that RAR alpha is not significantly involved in mediating retinoid-induced skin irritation in mice and suggest that RAR alpha-selective agonists may have reduced mucocutaneous side effects relative to other retinoids. PMID- 9334835 TI - Association of Directors of Anatomic and Surgical Pathology. Recommendations for the reporting of larynx specimens containing laryngeal neoplasms. AB - The Association of Directors of Anatomic and Surgical Pathology have developed recommendations for the surgical pathology report for common malignant tumors. The recommendations for carcinoma of the larynx are reported herein. PMID- 9334836 TI - Immunodetection of epithelial mucin (MUC1, MUC3) and mucin-associated glycotopes (TF, Tn, and sialosyl-Tn) in benign and malignant lesions of colonic epithelium: apolar localization corresponds to malignant transformation. AB - Epithelial mucins are present at the apical membranes of gastrointestinal epithelial cells or in their secretions. In this study, we examined the occurrence of peptide epitopes of the mucins MUC1 and MUC3 and of three mucin associated glycotopes (TF, Tn, and s-Tn) in a series of colorectal tissue samples (normal colon, adenomas with different grades of dysplasia, carcinoma in situ, and invasive carcinomas). A new monoclonal antibody to a conformation-dependent peptide epitope of MUC1 was employed, which does not react with the fully glycosylated mucin as found in normal gastrointestinal mucosa. We found that adenomas acquired the ability to expose Tn, s-Tn, TF and MUC1 epitopes, and this correlated with increasing malignant potential. The secretory mucin, MUC3, revealed a different pattern: it was detectable in all sections, with maximum expression in adenomas and decrease in carcinomas. Most importantly, normal mucosa and benign lesions showed supra-nuclear and/or apical distribution of these antigens, but malignant lesions and lesions with a very high risk of malignancy revealed diffuse cytoplasmic and basolateral membrane localization. The immunohistological response to a combination of MUC1-related antibodies may assist in assessing the malignant potential and status of lesions of the colon. PMID- 9334837 TI - Reciprocity between membranous and nuclear expression of beta-catenin in colorectal tumours. AB - beta-Catenin has a central role not only in linking the cadherin-mediated cell adhesion system but also in the intercellular signalling pathway. To investigate alterations of beta-catenin in the development of colorectal carcinoma, the pattern of beta-catenin expression was studied using immunohistochemistry in 74 sporadic colorectal adenomas, in histologically normal mucosa adjacent to 65 of these adenomas, and in 52 carcinomas arising in adenomas. All normal epithelia displayed cell boundary staining for beta-catenin. Adenomas and carcinomas showed varying degrees of membranous staining. However, some tumours also showed nuclear staining of beta-catenin protein. Decreased membranous and increased nuclear beta catenin staining were associated with increasing degrees of dysplasia in adenomas (P < 0.005, P < 0.05, respectively). Carcinomas manifested significantly reduced membranous, but enhanced nuclear beta-catenin expression compared with their associated adenomas (P < 0.001, P < 0.005, respectively). An inverse correlation was found between decreased membranous and increased nuclear staining of beta catenin in both adenomas and carcinomas (P < 0.025, P < 0.05, respectively). The data confirm that reduced membranous and increased nuclear expression of beta catenin is associated with the progression of colorectal adenomas to carcinomas. Our results also suggest that decreased membranous expression of beta-catenin may result from aberrant localisation of the protein in the cell nucleus. PMID- 9334838 TI - DNA aneuploidy, S-phase fraction, nuclear p53 positivity, and survival in non small-cell lung carcinoma. AB - Inactivation of the p53 gene plays a key role in tumour biology, probably through a disturbed cell cycle control and an increased genetic instability in p53 inactivated tumours. To learn more about the relationship between p53 alterations, proliferation and genetic instability (DNA aneuploidy) in lung cancer patients, specimens of 220 surgically resected lung carcinomas with clinical follow-up information were examined by immunohistochemistry (p53; CM1) and flow cytometry. Nuclear p53 positivity--found in 49.5% of the tumours--was associated with both high S-phase fraction (SPF) and DNA ploidy aberrations. SPF was higher in p53-positive tumours (15.9 +/- 10.2) than in p53-negative tumours (10.3 +/- 8.7; P = 0.03). The rate of p53 positivity was higher in 101 DNA aneuploid and DNA-multiploid tumours (55%) than in 27 diploid and peridiploid carcinomas (33%; P = 0.0512). These results are consistent with an in vivo role of p53 inactivation for increased proliferative activity and development of genomic instability in lung cancer. There was no association between SPF and prognosis. Although prognosis was worse in DNA-aneuploid and multiploid tumours than in diploid, peridiploid and tetraploid carcinomas (P = 0.029), DNA ploidy was not an independent predictor of poor prognosis in multivariate analysis. These data show that DNA-flow cytometry has little prognostic value for patients with resected non-small-cell lung carcinoma. PMID- 9334839 TI - Inflammatory markers in chronic hepatitis C. AB - To test the hypothesis that inflammation in hepatitis C follows mechanisms common to immune-activated pathways, the distributions of T and B cells, adhesion molecules and transforming growth factor-beta (TGF-beta) were assessed in liver biopsies with chronic inflammation due to hepatitis C (HCV, n = 8) and other causes (non-HCV, n = 10). Frozen sections were immunostained using primary antibodies to CD2, CD20, CD4, CD8, intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM)-1, HLA-DR, lymphocyte function-associated antigen (LFA)-1, and TGF-beta. Inflammatory cells positive for each immunophenotypic marker were counted, and positive staining for adhesion molecules, HLA-DR and TGF beta was graded in triads and lobules and compared in HCV and non-HCV biopsies. In all biopsies, T cells were more frequent than B cells, both in triads and lobules. CD20+, CD4+, CD8+ and LFA-1+ cells were increased in HCV compared to non-HCV biopsies. Portal lymphoid aggregates were present in 6 of 8 HCV biopsies and 3 of 10 non-HCV biopsies. Aggregates consisted of CD20+, CD4+, CD8+ and LFA-1+ cells, and ICAM-1 and VCAM-1 were increased. Sinusoidal lining cells in HCV biopsies and non-HCV biopsies with inflammation expressed HLA-DR, ICAM-1, and CD4. TGF-beta was increased in foci of necrosis. Inflammation in chronic HCV involves common immune-mediated cellular effector pathways and the inflammation in the portal triads represents aggregation of both T and B cells, mediated in part by upregulation of adhesion molecules on portal stromal cells; this is possibly in response to antigens draining from necroinflammatory foci in the lobules. TGF-beta is increased in active necroinflammatory foci, but not in portal lymphoid aggregates. PMID- 9334840 TI - Frequency and distribution of DNA fragmentation as a marker of cell death in chronic liver diseases. AB - To study the early stages of cell death in various types of chronic liver injury, liver biopsies from a total of 26 patients, including 7 with chronic hepatitis C(CHC), 4 with chronic hepatitis B(CHB), 7 with alcoholic liver disease (ALD), 4 with autoimmune or drug hepatitis (AI/DH), and 4 with primary biliary cirrhosis(PBC), were examined by an in situ nucleotidyl transferase assay (ISNTA), which detects DNA fragmentation. Positive nuclei in hepatocytes and sinusoidal lining cells were counted in all parenchymal areas, excluding triads and areas of fibrosis, using a computer with Sigmascan software. The number of positive hepatocytes/mm2 was similar in the biopsies of patients with CHC, CHB, ALD and AI/DH, but significantly lower in PBC. The number of positive sinusoidal lining cells/mm2 was significantly greater in biopsies with CHC compared to CHB, ALD, AI/DH and PBC. Double staining revealed that the ISNTA-positive sinusoidal lining cells were also CD68 positive, indicating that they were Kupffer cells. The frequency of ISNTA positivity did not correlate with serum AST or ALT levels, steatosis, cell swelling or cirrhosis. ISNTA-positive hepatocytes were more frequent than acidophilic bodies in every disease category. We conclude that apoptosis may be a common pathway of cell death in different liver diseases, that the high frequency of DNA fragmentation in Kupffer cells in CHC suggests that during chronic hepatitis C infection activated Kupffer cells may be subject to regulatory control by apoptosis and that ISNTA is more sensitive than acidophilic bodies in assessing the degree of cell injury in the liver. PMID- 9334841 TI - Expression of pancreatic digestive enzymes in normal and pathologic epithelial cells of the human gastrointestinal system. AB - Pancreatic digestive enzymes have rarely been reported in human nonpancreatic organs. We examined their expression in the epithelial cells of the nonpancreatic gastrointestinal organs, looking for pancreatic alpha-amylase, trypsin, chymotrypsin and pancreatic lipase. Western blotting, enzyme assay and pancreatic alpha-amylase mRNA were also used in selected specimens. In normal tissues, immunoreactivity of one or more of these enzymes was frequently noted in cells of the salivary glands, stomach, duodenum, large pancreatic ducts, extrahepatic bile ducts and gall bladder. The epithelium of the normal oesophagus, small intestine and colon were consistently negative for these enzymes. In pathologic tissues, immunoreactivity for one or more enzymes was present in epithelial cells of pleomorphic adenomas of the salivary glands, oesophageal squamous cell carcinoma, gastric adenoma and adenocarcinoma, pancreatic adenocarcinoma, cholecystitis, adenocarcinoma of the gall bladder and extrahepatic bile duct, and colon adenoma and adenocarcinoma. Western blotting showed a specific band of each enzyme in some specimens of normal stomach. In situ hybridization for pancreatic alpha amylase mRNA showed specific signals in the normal stomach, but not in the normal colon. Reverse transcriptase polymerase chain reaction analysis for pancreatic alpha-amylase mRNA revealed specific signals in the normal stomach. Enzyme assay revealed that the stomach and gall bladder showed these activities. The data suggest that pancreatic digestive enzymes are produced by several epithelial cell types of the nonpancreatic gastrointestinal organs, that the organs positive for pancreatic enzyme have a common cell lineage, and that neoplasms continue to express or neoexpress these enzymes after neoplastic transformation. PMID- 9334842 TI - Apocrine epithelium of the breast: does it result from metaplasia? AB - Benign and malignant breast lesions may show an apocrine epithelium considered to be the result of metaplasia. In an attempt to clarify the histogenesis of the breast apocrine epithelium we searched for the presence of apocrine cells or cells with apocrine differentiation during human breast development. We analysed 10 autopsy specimens of female breasts from fetuses between 28 and 40 weeks of gestational age and tissue from 6 normal breasts, obtained after breast reduction in nulliparous young women between 22 and 28 years of age. Formalin-fixed, paraffin-embedded sections were stained with haematoxylin-eosin, PAS-diastase and a monoclonal antibody (BRST-2) anti-GCDFP-15, which is a specific apocrine marker. A 40-week fetal breast was analysed by electron microscopy. No cells with histological and ultrastructural apocrine features were found in the ducts of fetal breasts. All fetal breasts showed some ducts with sparse GCDFP-15 immunoreactive cells; the number of these cells increased with gestational age. PAS-diastase was negative. No cells with apocrine morphology were found in ducts and lobules of normal adult breasts. Scattered GCDFP-15-positive luminal epithelial cells and rare PAS-diastase-positive cells were observed in some lobules of all adult breasts. Cells with biochemical characteristics (GCDFP-15 expression) of apocrine differentiation are evident during human fetal breast development and persist in adult mammary glands. Unknown stimuli may induce these cells to take on an apocrine morphology. Apocrine epithelium of the breast may be a normal process of differentiation rather than metaplasia. We suggest the term "apocrine differentiation precursor cells" for GCDFP-15-positive breast epithelial cells with no apocrine morphology. PMID- 9334843 TI - Metastatic carcinoma of presumed prostatic origin in cremated bones from the first century A.D. AB - A cremated pelvis dating from the first century A.D. showed evidence of osteosclerotic metastasis, presumably secondary to prostate carcinoma. The case demonstrates the importance of microradiography in palaeopathology as well as some of the structural changes seen in cremated bone. PMID- 9334845 TI - A case of inflammatory pseudotumour of the common bile duct. AB - Inflammatory pseudotumour of the common bile duct (CBD) is extremely rare. A 58 year-old Japanese female without choledocolithiasis underwent pancreatico duodenectomy for constriction of the middle lower region of the CBD. A submucosal tumour protruding into the CBD, was histologically inflammatory consisting of fibroblastic cells, collagen fibres and myxoid stroma with chronic inflammatory cells. This lesion was surrounded by an irregular fibrosclerosing lesion with obliterative phlebitis which involved the neighbouring pancreas and lymph nodes. Clonal analysis of the tumour by polymerase chain reaction analysis of X chromosome inactivation patterns, confirmed the polyclonal nature of the lesion. Immunohistochemically, the fibroblastic cells in both lesions had the same phenotype [vimentin (+), desmin (-), muscle-specific actin (-) and CD34 (+)] suggesting that these lesions with different histological features represent zonation of the same inflammatory process. The outer lesion extended irregularly into adjacent pancreatic tissue and lymph nodes. This fact made it difficult to differentiate this from a malignant lesion, even if frozen sections contained no atypical cells. PMID- 9334844 TI - Osteoclast-like giant cell tumour of the pancreas presenting as a pseudocyst-like lesion. AB - A 57-year-old male patient presented with a cystic lesion in the tail of the pancreas, which was considered to be a pseudocyst. He was treated by cystojejunostomy but one year later a tumour was found to have invaded the stomach and jejunum. This was an osteoclast-like giant cell tumour containing a small area of typical ductal adenocarcinoma. Immunohistochemical staining revealed that the pleomorphic tumour cells were positive for cytokeratin, epithelial membrane antigen, vimentin and the proliferation marker MIB-1. The osteoclast-like giant cells and some small histiocytic cells stained for leukocyte common antigen and histiocytic markers and were negative for MIB-1. At autopsy, tumour rests were found in the pancreas but there were no metastases. Osteoclast-like giant cell tumours of the pancreas may present as cystic lesions and should be included in the differential diagnosis of pseudocysts. PMID- 9334850 TI - Type I interferons enhance production of IFN-gamma by NK cells. AB - In murine models, challenge with different viral and parasitic infection is closely associated with the production of type I interferons (IFN-alpha/beta) and NK cell production of interferon-gamma (IFN-gamma). Therefore, we wished to determine if IFN-alpha/beta had a role in the regulation of NK cell production of IFN-gamma. IFN-alpha/beta alone stimulated low levels of IFN-gamma production by purified populations of IL-2 activated NK cells but in combination with IL-12 resulted in the production of significant levels of IFN-gamma. Interestingly, maximal production of IFN-gamma by NK cells stimulated with IL-2 plus IFN alpha/beta was dependent on endogenous tumor necrosis factor-alpha (TNF-alpha). Further studies revealed that TNF-alpha enhanced the ability of IFN-alpha/beta to stimulate production of IFN-gamma by NK cells. In contrast to the stimulatory effect of IFN-alpha/beta on NK cell production of IFN-gamma, IFN-alpha/beta inhibited IL-2 induced proliferation of NK cells. This inhibitory effect was not reversed by the addition of neutralizing antibodies specific for IFN-gamma or TNF alpha. These data demonstrate that the type I interferons enhance NK cell production of IFN-gamma and suggest that they may be important in the regulation of NK cell production of IFN-gamma during infection. PMID- 9334851 TI - Inhibitory effect of nicotinamide on in vitro and in vivo production of tumor necrosis factor-alpha. AB - Nicotinamide, a pellagra-preventive factor, has multiple functions such as inhibition of poly-ADP-ribose synthetase, inhibition of inducible nitric oxide synthase, free radical scavenging and suppression of major histocompatibility complex class II expression and ICAM-1 expression on endothelial cells. In addition to these, we have found an inhibitory effect of nicotinamide on production of tumor necrosis factor-alpha (TNF-alpha) in vitro and in vivo. Lipopolysaccharide (LPS)-induced in vitro TNF-alpha production by human peripheral blood mononuclear cells, measured by enzyme-linked immunosorbent assay (ELISA), was significantly inhibited with more than 1 x 10(-3) mol/l of nicotinamide, while interleukin-1-beta was not inhibited and interleukin-6 was slightly inhibited even with 10(-2) mol/l. Oral administration of nicotinamide with more than 62.5 mg/kg also significantly inhibited LPS-induced serum TNF alpha production measured by ELISA and bioassay in Balb/c mice. Thus, nicotinamide has an inhibitory effect on TNF-alpha production that may be beneficial to TNF-alpha-mediated diseases. PMID- 9334852 TI - Genetic regulation of the impaired immune response to hepatitis-B vaccine associated with low TCR density in end stage renal disease patients: contribution of complement C4 and factor B alleles. AB - We have studied the relationship between T-cell receptor (TCR) density, genetic factors and the specific immune response in 153 end stage renal disease (ESRD) patients on haemodialysis immunised with HBsAg vaccine. One-hundred and nineteen patients raised a protective (> 10 U/ml) antibody response to hepatitis-B vaccination (responder, R), while 34 patients were found to be non-responders (NR). The density of the T-cell receptors was determined by flow cytometry. Proliferation of the T-cells induced by autologous monocytes presenting HBsAg was also measured and expressed as a stimulation index (SI). MHC class I, II and III alleles of the patients were also determined. The densities of TCR/CD3 receptors in NR patients were found to be significantly decreased as compared to the R patients (189 +/- 22 vs. 282 +/- 58 arbitrary units, P = 1.3 x 10(-7). TCR/CD3 receptor densities were found to be strongly associated (Spearman correlation coefficient: 0.84, P < 0.000001) with the SI values. Both parameters were found to be under dual genetic control: (a) very low density of the TCR/CD3 receptors and very low SI were found mainly in NR patients carrying HLA-A1, HLA-B8 and HLA DR3 alleles; and (b) TCR/CD3 densities and function in R group were found to be significantly lower in carriers than in non-carriers of two MHC class III complement protein alleles: C4A*6, and Bf*F. Non-responsiveness to hepatitis-B vaccination was found to be associated with extremely increased neopterin levels. These findings indicate that both genetic and acquired factors contribute to the hepatitis-B vaccination failure in ESRD patients. PMID- 9334853 TI - IL-7 induces proliferation, variable cytokine-producing ability and IL-2 responsiveness in naive CD4+ T-cells from human cord blood. AB - We investigated the effects of IL-7 on the proliferation and acquisition of cytokine-producing ability of naive CD4+ T-cells from human cord blood. Naive CD4+CD45RA+ T-cells from human cord blood expressed CDw127 (IL-7R) at higher levels than adult CD4+ CD45RA+ T-cells and produced IL-2 and a small amount of IFN-gamma upon stimulation with PMA and ionomycin. IL-7 induced IL-2-independent proliferation and both Th1- and Th2-type cytokine-producing abilities in cord blood CD4+CD45RA+ T-cells without stimulation via TCR. These results suggest that this IL-7-induced antigen-independent activation mechanism could contribute to maintaining the clonal size of naive T-cells with the potential to differentiate into either Th1 or Th2 cells at the sites of IL-7-expression. PMID- 9334854 TI - Effects of hCG and beta-hCG on IL-2 and sIL-2R secretion from human peripheral blood mononuclear cells: a dose-response study in vitro. AB - The effects of human chorionic gonadotropin (hCG) as well as beta-subunit of hCG (B-hCG) in concentrations of: 80,000/25,000; 500; 50; 5 mIU/ml on in vitro release of interleukin-2 (IL-2) and soluble interleukin-2 receptor (sIL-2R) from resting or phytohemagglutinin (PHA) activated human peripheral blood mononuclear cells (PBMC) were studied. Both the interleukins were measured in supernatants of human PBMC by quantitative sandwich enzyme immunoassay method (ELISA). We found that hCG in the dilutions of 80,000 mIU/ml (P < 0.01) and 5 mIU/ml (P < 0.05) diminished IL-2 secretion only from PHA activated PBMC. beta-hCG in concentrations of 5 mIU/ml (P < 0.05), 500 mIU/ml (P < 0.01) and 25,000 mIU/ml (P < 0.05) also diminished IL-2 secretion from PHA activated PBMC, and only in a dilution of 25,000 mIU/ml (P < 0.05) from resting PBMC. Simultaneously, hCG in concentration of 80,000 mIU/ml (P < 0.01) potentiated the release of sIL-2R into supernatants from resting and PHA activated PBMC, but in concentration of 50 mIU/ml (P < 0.05) slightly depressed the secretion of IL-2 from PHA activated PBMC cultures. beta-hCG in dilution of 25,000 mIU/ml (P < 0.001) stimulated the release of sIL-2R from resting or PHA activated PBMC. beta-hCG had also inhibitory effect on sIL-2R secretion from resting (in a dilution of 50 mIU/ml; P < 0.05) and PHA activated (500 mIU/ml; P < 0.01) PBMC. The inhibitory effect of very high concentrations of hCG and beta-hCG on IL-2 secretion together with their stimulatory effect on sIL-2R release from PBMC may be an important event during the human pregnancy and various cancers. PMID- 9334856 TI - Apoptotic and necrotic cytolytic processes induced by membrane associated proteins of LAK cells. AB - Cytolytic processes induced by membrane-associated proteins of human lymphokine activated killer (LAK) cells with different phenotypes (CD3+, CD16-, CD8+ and CD16+, CD8+, CD3-) were studied using L929 and K562 types of target cells. Independently of the phenotype of effector cells and the type of target cells, total fractions of membrane proteins induced several different cytolytic processes occurring with different rates and involving different mechanisms of genome fragmentation. The membrane fraction induced, irrespective of the phenotype of LAK cells, mostly apoptotic processes in the L929 line. At the same time, cytolytic processes induced in K562 line differed by the mechanisms of DNA fragmentation. An inhibitor of lysosome activation, NH4Cl, and a Ca(2+)-binding reagent, ethylene glycol bis-(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), induced partial inhibition of short-term cytolytic processes (developing within 1-5 h) but did not affect the development of long-term cytolytic processes requiring more than 6-8 h for their development. PMID- 9334855 TI - Soluble haemoglobin is a marker of recent Plasmodium falciparum infections. AB - Monoclonal antibodies (Mab) were raised against haemoglobin (Hb) associated with Plasmodium falciparum protein and used to develop an ELISA, measuring circulating levels of released Hb. This assay was evaluated in different malaria patients in parallel with ELISA assays for C-reactive protein (CRP) and haptoglobin. Levels of Hb were negatively associated with levels of haptoglobin. Increased levels of serum Hb and CRP and decreased levels of haptoglobin were seen in Danish malaria patients. Consecutive studies showed that increased Hb levels were detectable 3-7 days after initiation of treatment probably because of drug induced destruction of infected erythrocytes. Increased levels of CRP were measured 0-3 days after initiation of treatment. The Hb assay was used in an epidemiological study of malaria in an area of Sudan with unstable malaria transmission. The proportion of Sudanese adults with detectable soluble Hb was higher in the rainy season with malaria transmission compared to the dry season. Hb levels in the rainy season were negatively associated with levels of haptoglobin. Most adults had increased levels of soluble Hb and decreased levels of haptoglobin 7 and 30 days after their treatment of P. falciparum malaria compared to the levels during acute disease. Thus, both soluble Hb and haptoglobin appear to be markers of recent P. falciparum infections. Very high levels of CRP protein were measured in some of the malaria patients at the day of treatment while lower levels were recorded 7 and 30 days after treatment. Soluble Hb levels were associated with malariometric parameters in a similar fashion to haptoglobin. The new Mab-based assay for measuring soluble Hb in the peripheral blood of malaria patients may be useful for future epidemiological studies of malaria. PMID- 9334857 TI - Fucoidin, a potent inhibitor of L-selectin function, reduces contact hypersensitivity reaction in mice. AB - In order to investigate the role of L-selectin (CD62L) in delayed-type hypersensitivity (DTH) reaction, effect of fucoidin, a potent inhibitor of CD62L function, was examined in a model of mouse contact hypersensitivity reaction. Intravenous injection of fucoidin to sensitized mice just before hapten challenge resulted in a significant and dose-dependent reduction of the ear swelling in contact hypersensitivity. The ear swelling caused by the hapten challenge was also inhibited when fucoidin was administered at the sensitization phase. Histological analyses of the ear sections revealed that the fucoidin-induced suppression of contact hypersensitivity reflected a marked inhibition of the ear edema and the leukocyte infiltration. The activity of fucoidin was specific in that its related saccharides exerted little effect on the reaction. These results suggest that CD62L may play an important role in both afferent and efferent phases of cutaneous DTH reaction. Since DTH response is one of the most significant features of several chronic inflammatory diseases, our data also show that blocking of CD62L function may be beneficial for the treatment of these diseases in humans. PMID- 9334859 TI - Will cultures survive? The role of molecular tests in diagnostic bacteriology. PMID- 9334858 TI - Inhibition of etoposide-induced apoptotic events by azide. AB - Azide is reported to be an inducer of necrotic rather than apoptotic cell death. Using various parameters of cell death, we demonstrate here that azide is capable of completely inhibiting apoptosis induced by VP-16/etoposide. Azide was found systematically to protect Jurkat cells against VP-16-induced effects as determined by an array of biochemical and morphological parameters. The cells were able partially to recover proliferative activity following removal of the drugs. We conclude that azide inhibits apoptosis induced by VP-16 at an early point in the apoptotic pathway and that inhibition of apoptosis might be the mechanism of necrosis-induction by azide. PMID- 9334860 TI - Prophylaxis of cytomegalovirus disease in high-risk patients. PMID- 9334861 TI - Tuberculosis and HIV infection: a review. PMID- 9334862 TI - Atherosclerosis due to chronic arteritis caused by Chlamydia pneumoniae: a tentative hypothesis. PMID- 9334863 TI - Vaginal Ureaplasma urealyticum colonization: influence on pregnancy outcome and neonatal morbidity. AB - The effects of Ureaplasma urealyticum colonization on pregnancy and neonatal outcome was prospectively studied in women with impending term or preterm delivery. One hundred and seventy women colonized with U. urealyticum as the only pathogenic microorganism and 83 women with negative cultures were enrolled for study. Compared to the controls, U. urealyticum colonization was associated with a significantly increased rate of amnionitis (2% vs 35%; p < 0.001), chorioamnionitis (0% vs 10%; p < 0.05), premature rupture of membranes (12% vs 35%; p < 0.001) and preterm delivery (10% vs 41%; p < 0.001). The rate of vertical transmission ranged from 38% in term infants to 95% in very low birth weight infants. U. urealyticum colonization at birth was associated with an increased risk for the development of respiratory distress syndrome (9% vs 51%), intraventricular hemorrhage (1% vs 7%) and bronchopulmonary dysplasia (4% vs 17%) in very low birth weight infants (< 1500 g). It is concluded that maternal U. urealyticum colonization is associated with amnionitis, chorioamnionitis and preterm delivery, and that tracheal colonization with U. urealyticum increases the risk for respiratory and neurological complications in very low birth weight infants. PMID- 9334864 TI - Relationship between the Borrelia burgdorferi specific immune response and different stages and syndromes in neuroborreliosis. AB - One hundred untreated neuroborreliosis patients were investigated by IgG/IgM immunoblot to find out if different stages and syndromes are characterized by different patterns of their Borrelia burgdorferi specific immune responses in CSF and serum. Stage III (n = 10) was characterized by a broad, highly specific intrathecal immune response (the mean number of IgM bands in CSF was 3.1 and of IgG bands 6.3). All patients recognized one or more of the following proteins (p35, p21, p18) or the 5 kd glycolipid. In contrast, the immune response in stage II (n = 90) was less restricted (28%) and heterogeneous (mean number of IgM bands 1.4 and of IgG bands 3.4). It was mainly directed against the highly crossreactive p41 antigen (91%). The different clinical features of stage II were comparable regarding the intrathecal immune response. However anti-glycolipid and anti-p35, -p21, -p18 IgG antibodies were detected in a small subset of patients, mainly corresponding to more severe courses of the disease. Our data are compatible with a direct agent-related pathomechanism in neuroborreliosis. Antibodies against certain proteins and the glycolipid of B. burgdorferi seem to have a prognostic value as to the development of more severe disease or transition to stage III. PMID- 9334865 TI - Immunogenicity and reactogenicity of HbOC vaccine administered simultaneously with acellular pertussis vaccine (DTaP) into either arms or thighs of infants. AB - To evaluate the reactogenicity and immunogenicity of a Haemophilus influenzae type b conjugate vaccine (HbOC) and of a tricomponent acellular pertussis vaccine (DTaP) when injected simultaneously into either contralateral arms or into contralateral thighs, 110 infants were enrolled to receive three doses of DTaP at 3, 4, and 5 months and two HbOC doses at 3 and 5 months of age. Administration of either of the two vaccines into arms was associated with significantly more local side effects than administration into thighs. There was no difference in geometric mean concentration (GMC) values for any of the four vaccine antigens between subjects who had been vaccinated into arms or thighs. After immunization, all children had protective antibody titers to diphtheria toxin. While post vaccination the mean anti-tetanus toxoid GMC was > or = 1.25 IU/ml, there was no significant rise as compared to the GMC before vaccination. GMCs of antibodies against the various pertussis antigens were similar to those observed before with the same DTaP vaccine. The simultaneous administration of DTaP and HbOC was safe and immunogenic irrespective of the site of vaccine administration, but significantly more local reactions occurred when vaccines were injected into arms. PMID- 9334866 TI - Antifungal activity of ketoconazole and other azoles against Malassezia furfur in vitro and in vivo. AB - The in vitro activity of several antifungal agents (ketoconazole, miconazole, econazole, fenticonazole, itraconazole, fluconazole) in routine clinical use against Malassezia furfur infections has been studied with freshly isolated strains of M. furfur from pityriasis versicolor lesions. The results indicate that the drugs tested exert a good activity, and both ketoconazole and itraconazole appear very active (0.8 mg/l respectively). Hair samples from the beards of volunteer patients affected by pityriasis versicolor but otherwise healthy were examined to determine ketoconazole levels during oral therapy (one or two 200 mg tablets daily). It was shown that the drug progressively accumulates in the beard, reaching levels proportional to the dose administered, although blood levels did not increase in parallel. The study of drug concentration profile has evidenced a long ketoconazole persistence in the beard at therapeutic levels. In conclusion, the possibility of reaching high and lasting ketoconazole levels in the keratin layer of the epidermis indicates that systemic ketoconazole therapy could be useful for eradication of M. furfur in patients affected by pityriasis versicolor. PMID- 9334867 TI - Prevalence of serum HGV-RNA among hemodialysis patients in Turkey. AB - A possible agent for human non-A-E hepatitis has been identified and named hepatitis G virus (HGV). The aim of this study is to evaluate the prevalence of serum HGV-RNA among hemodialysis patients in our country and the possible correlations of serum HGV-RNA with antibody to hepatitis C virus (anti-HCV), chronic liver dysfunction, number of blood transfusions, serum hepatitis B surface antigen (HBs Ag), duration of hemodialysis therapy, history of renal transplantation and patients' age and sex. Seventy-eight hemodialysis patients and 59 healthy controls were included in the study. Twenty-seven of 78 hemodialysis patients (34.6%) and two of the 59 healthy controls were serum HGV RNA positive (p < 0.01, x2 = 17.8). There was no significant difference between the HGV-RNA positive and HGV-RNA negative groups regarding mean duration of dialysis therapy, anti-HCV, chronic liver dysfunction, number of blood transfusions, serum HBs Ag, duration of hemodialysis therapy, history of renal transplantation and patients' age and sex. In conclusion, hemodialysis patients carry the risk for HGV infection and transmission routes and clinical significance of HGV infection in these patients remain to be defined. PMID- 9334868 TI - Primary herpes simplex virus type 1 gingivostomatitis in pediatric personnel. AB - Herpetic gingivostomatitis is common in young children, but primary oral infection has also been described in adults. Herpetic whitlow as an occupational hazard of medical personnel has been well documented. Four cases of primary herpetic gingivostomatitis are reported in two pediatricians and two pediatric nurses who contracted the infection in their fourth decade of life. All suffered from sore throat and fever as presenting symptoms. Correct diagnosis was delayed for 4-5 days. In conclusion, pediatric personnel with pharyngitis and a negative history of herpetic gingivostomatitis or herpes labialis should bear the possibility of oral HSV infection in mind. Early diagnosis is essential to prevent the spread of the infection to their patients. PMID- 9334869 TI - Acute hepatitis E in Catania (eastern Sicily) 1980-1994. The role of hepatitis E virus. AB - Between 1980 and 1994, 540 patients with acute viral hepatitis were admitted to hospital at the Department of Infectious Diseases of Catania (eastern Sicily). Twenty-five patients out of 540 were assessed as having non-A, non-B, non-C hepatitis. These subjects were studied for anti-HEV IgM and IgG seroprevalence by testing serial serum samples collected 1, 4, 12 and 24 weeks after the onset of acute disease. Fourteen of 25 samples (56%) seroconverted to anti-HEV IgG antibodies. No sample was positive for anti-HEV IgG at week 1, ten samples were positive at week 4 and the remainder at week 12. Anti-HEV reactivity was maintained until week 24 in all cases. In 11 of the 14 patients seroconverting to anti-HEV, the presence of IgM anti-HEV was found, which appeared in the sample from week 1 and gradually disappeared thereafter. Identified risk factors for HEV transmission included travel in the tropics and shellfish ingestion (anti-HEV positive versus anti-HEV negative: p < 0.05). HEV-related hepatitis is not yet a major public health problem in Sicily but, from our data, the trend of its incidence is clearly upwards. The high incidence of faecally-orally transmitted diseases in Sicily, the crucial position of Sicily in the middle of the Mediterranean Sea (where HEV largely circulates) and the increase of migration from developing countries are all factors which should increase awareness for a more active surveillance of the spread of HEV in our area. PMID- 9334870 TI - Appendicitis followed by reactive arthritis in an HLA B27-positive man after infection with Yersinia enterocolitica, diagnosed by serotype specific antibodies and antibodies to Yersinia outer membrane proteins. AB - A case of appendicitis followed by reactive arthritis in an HLA B27-positive, 29 year-old man after infection with Yersinia enterocolitica is reported. Infection with Y. enterocolitica was diagnosed by determination of serotype specific antibodies and antibodies to Yersinia outer membrane proteins. Bacteriological cultures from the appendix were not made. Although reactive arthritis is a well known complication of Yersinia-associated enteric disease, there are only few reports of patients with Y. enterocolitica pseudo-appendicitis complicated by arthritis during follow-up. PMID- 9334871 TI - Malignant boutonneuse fever and polymyalgia rheumatica: a coincidental association? AB - Mediterranean spotted fever is a tick-borne disease that is endemic in the Mediterranean basin from spring to autumn. Usually mild, the disease can be severe in some cases, especially when risk factors are encountered in patients or when treatment is delayed. The correlation between these malignant forms and patients' immunological disorders remains unclear, while the pathophysiology of the disease seems well known. A case of a malignant form of Mediterranean spotted fever is reported which occurred 2 months prior to the diagnosis of polymyalgia rheumatica. Evidence of immunological disorders consisted only in an antiphospholipid antibody associated with a transient lupus anticoagulant. No underlying risk factors other than the primary undiagnosed phase of polymyalgia rheumatica has been observed. PMID- 9334872 TI - Incidence and risk factors of methicillin-resistant Staphylococcus aureus colonisation/infection in an intensive care unit. PMID- 9334873 TI - Toscana virus infection in German travellers returning from the Mediterranean. PMID- 9334874 TI - HHV-8 in PBMC and Kaposi's sarcoma activity. PMID- 9334875 TI - Intergenerational perceptions of English speaking and Spanish speaking Mexican American grandparents. AB - Hispanics are facing a number of problems, such as poverty, hunger, and a high dropout rate at school. Health-care reform and changes in Medicaid and Medicare are bound to further challenge the resiliency of minority families. To strengthen families from within, relevant programming should be implemented. Information regarding the strengths and needs of Mexican-American grandparents was obtained in order to adapt existing grandparenting programs for this population. Mexican American grandparents (n = 181), parents (n = 148), and grandchildren (n = 173) provided information on grandparent satisfaction, success, teaching, difficulty, frustration, and information needs. Multivariate analyses of variance found differences for English and Spanish speaking grandparents. Spanish speaking grandparents reported a greater need for information than English speaking grandparents, and more frustration when dealing with adolescents than with younger children. For the English speaking grandparents, all of the generations agreed that grandparents under the age of sixty-one experienced more frustration than their older counterparts, and those who spent more than five hours a month with their grandchildren were more effective in their role. Possible factors that account for the findings are discussed and recommendations for establishing a grandparent program are presented. PMID- 9334876 TI - Granddaughters' accounts of the influence of parental mediation on relational closeness with maternal grandmothers. AB - This study explores granddaughters' reports of the nature and impact of parental mediation on the development of their feelings of closeness toward their maternal grandmothers. Results from close-ended questions assessing granddaughters' perceptions of parental mediation and relational closeness indicated that closeness was predicted by mediation involving: visits alone with grandmothers, the absence of criticism of grandmothers, communication indicating the grandmothers were important persons, and less influence on conversations or activities. Qualitative analysis of granddaughters' accounts of the influence of parents illustrated what they perceived to be important issues in mediation. Additionally, accounts revealed the role of other family members in the relationship mediation process. PMID- 9334877 TI - Unanticipated separations and normative re-attachments in later life: a personal account. AB - Unanticipated separations can have profound reverberations upon individual and family development. When these separations are at a young age the consequences tend to exert a long and lasting effect. This article presents a personal account of the abrupt separation from the author's parents, the trauma the family faced in Europe on the brink of war and how this impacted his family intergenerationally. It is suggested that loyalties and caregiving are dynamics which enable adult children to emotionally reconnect with their adult parents in later life. These renewed levels of attachments are normative in nature, enriching the family as a whole. PMID- 9334878 TI - The therapeutic role in later life: husbands, wives and couples. AB - The present study investigates the extent to which marital partners are different or similar in their ways of enacting the therapeutic, or supportive, role. Specifically, the article compares husbands and wives categorically as in non dyadic studies and then as marital partners as in dyadic studies. In addition, this study, by using data from the Aging Couples Study, included only dual-earner couples so as to control for the effects of work life on marital relations. Results showed that studies of individual married men and women understate the differences between marital partners in that some wives "overbenefit" in the exchange of conjugal supports. However, husbands more often "overbenefit." Findings also indicated that the norm of reciprocity does not prevail regarding the extent of support, although it does for the types of support exchanged. PMID- 9334879 TI - Biographical reconstruction of WWII experience: an exploration of German remembrance. AB - First results of the Eldermensch-Study (N = 50) indicate that German National Socialism and the historical events associated with World War II play a central role in the oral histories of older Germans. Six distinct patterns of late life cognitive appraisal of war-time experience: survival challenge (further divided into "active" and "passive" survivorship), passive benefit, opportunism, value verification, and acceptance despite hardship are presented and discussed as structural themes in the life story. These categories not only represent interindividual differences in the appraisal of war-time experience, but often reflect contrasting patterns of personal response to the social and political circumstances of this era of German history. In addition to cohort and gender differences, divergence in locus of control orientation as well as the role of social bonding and instrumental social support are discussed. Differences revealed in the biographical interviews are further reflected in the results of standardized questionnaires. PMID- 9334880 TI - Is guanosine-5'-triphosphate involved in calcium-activation of contractile proteins in vascular smooth muscle? AB - Isometric tension was measured to investigate the effects of guanosine-5' triphosphate (GTP) on the run-down of myofilament Ca2+ sensitivity in isolated rat mesenteric arteries permeabilized with beta-escin. The Ca2+ sensitivity assessed by the EC50 value for the Ca2+ (0.1-100 microM)-tension relationship progressively runs down in the control strips, while it was well-preserved for 5 successive Ca2+ applications in the presence of GTP (50 microM); no significant difference was found in the Ca2+ sensitivity observed with the 1st Ca2+ application between the control and GTP-treated strips. Guanosine-5'-(2-O-thio) diphosphate (GDP beta S, 100 microM) significantly decreased the Ca2+ sensitivity with the 1st Ca2+ application and eliminated the run-down of Ca2+ sensitivity. GTP (3-150 microM), applied to the strips submaximally precontracted with Ca2+, had a little effect on the Ca2+ contractions in the early stage of experiments, but dramatically enhanced the Ca2+ contractions in their later stage; its latter effect was mimicked by guanosine-5'-(3-O-thio) triphosphate (GTP gamma S) and reversed by GDP beta S (100 microM). The results suggest: 1) loss of endogenous GTP following permeabilization is involved in the run-down of Ca2+ sensitivity; and 2) activation of G-proteins is involved in Ca(2+)-activation of contractile proteins. PMID- 9334881 TI - Disposition of intravenous theophylline in asthmatic children: Bayesian approach vs direct pharmacokinetic calculations. AB - Fifteen children (mean age +/- SD: 6.4 +/- 3.4, range: 2-12 years) with an acute asthma attack were treated by an intravenous dosage regimen of theophylline (30 min loading infusion of 6 mg/kg body weight followed by a constant infusion of 1 mg/kg, twice for 6 hr each). Three blood samples were drawn (each 15 min after the bolus infusion and after the two infusion periods of 6 hr). Plasma clearance (CL), apparent volume of distribution (Vd) and elimination half-life (t1/2) were estimated by the Bayesian approach using either only the first peak level (Bay 1) or all three monitored concentrations (Bay 3). These values were compared to the parameters calculated by a standard pharmacokinetic procedure (SC). Therapeutic steady state plasma levels around 12 micrograms/ml were rapidly achieved, and the pharmacokinetic parameters (CL = 1.1-1.5 ml/min/kg, Vd = 0.44-0.50 l/kg, t1/2 = 3.5-5.4 hr) differed slightly between the 3 methods applied. There was a significant linear correlation between the Bayesian-derived and SC-derived pharmacokinetic parameters. However the method Bay 1 seems to overestimate the elimination rate of theophylline more than Bay 3 does. In conclusion, Bayesian based therapeutic plasma level monitoring (Bay 3 are better than Bay 1) can be utilized for individualized pharmacokinetic calculations and proper dosage predictions of theophylline in pediatric patients. PMID- 9334882 TI - Effects of sucralfate and its components on acid- and pepsin-induced damage to rat gastric epithelial cells. AB - We have established models of cell damage induced by acid and pepsin using rat gastric epithelial cells (RGM1). In the present study, the effects of aluminum hydroxide [Al(OH)3] and potassium sucrose octasulfate (KSOS), which are components of sucralfate, and sucralfate on cell damage and peptic activity of pepsin were examined. Pretreatment of cells with sucralfate (0.1-3 mg/ml) or Al(OH)3 (0.1-1 mg/ml) for 2 hr prevented both acid- (pH 4.0) and pepsin- (pH 4.5) induced cell damage. However, KSOS (0.1-1 mg/ml) did not show any effects on two different types of cell damage. The peptic activity of pepsin at pH 4.5 was about 10% of that at pH 2.0. Sucralfate and KSOS slightly inhibited peptic activity at pH 4.5. Al(OH)3 inhibited peptic activity by approximately 50%; however, no concentration-dependent pattern was observed. Pepstatin (0.003-0.1 mg/ml), a specific inhibitor of pepsin, inhibited the peptic activity in a concentration dependent manner. Here, we confirmed that sucralfate and Al(OH)3 have cytoprotective effects against acid- and pepsin-induced cell damage. The mechanism behind the cytoprotective effects of sucralfate seems to relate to adhesion of the cell surface and neutralization of hydrogen ion by aluminum that prevents the penetration of hydrogen ions into the cells. PMID- 9334883 TI - Cilnidipine attenuates renal nerve stimulation-induced renal vasoconstriction and antinatriuresis in anesthetized dogs. AB - We examined the effects of cilnidipine, which is an L- and N-type Ca2+ channel blocker, on adrenergically regulated renal functions in anesthetized dogs. Renal nerve stimulation (RNS) at high frequency (3-7 Hz) decreased renal blood flow (RBF) without changes in systemic blood pressure. The RBF response was inhibited by intrarenal arterial (i.r.a.) infusion of cilnidipine at 0.1-0.3 microgram/kg/min. Low-frequency RNS (0.5-1 Hz) reduced absolute and fractional urinary sodium excretion. These responses were attenuated during i.r.a. infusion of cilnidipine at 0.3 microgram/kg/min. An increase in norepinephrine secretion rate induced by low-frequency RNS was also attenuated during cilnidipine infusion. These results suggest that cilnidipine can suppress norepinephrine release from the renal nerve endings and thereby interfere with the neural control of renal functions. PMID- 9334884 TI - Calcium oscillations in single cultured Chinese hamster ovary cells stably transfected with a cloned human cholecystokinin (CCK)B receptor. AB - In Chinese hamster ovary cells stably expressing the cloned human cholecystokinin (CCK)B/ gastrin receptor, cholecystokinin octapeptide (CCK-8) evoked increases in [Ca2+]i monitored by digitized video imaging of fura-2 fluorescence ratios. At concentrations around 10 pM, CCK-8 elicited [Ca2+]i oscillations, which were blocked by elimination of extracellular Ca2+, by a phospholipase C inhibitor, U 73122, by a protein kinase C inhibitor, H7, as well as by phospholipase A2 (PLA2) inhibitors, ONO-RS-082 and aristolochic acid. At higher concentrations, CCK-8 induced a single biphasic [Ca2+]i rise consisting of a large peak followed by a lower sustained plateau, while the response turned into [Ca2+]i oscillation when the extracellular Ca2+ was eliminated or a PLA2 inhibitor was included. CCK-8 stimulated the release of arachidonic acid, and this was inhibited by aristolochic acid. Arachidonic acid caused an increase in [Ca2+]i which was dependent upon extracellular Ca2+. These results suggest that the activation of PLA2 might be involved, at least in part, in the Ca2+ influx that maintains the sustained plateau phase of [Ca2+]i as well as the [Ca2+]i oscillation when CCKB receptors are stimulated. PMID- 9334885 TI - Pharmacological studies on the novel antiallergic agent HSR-609: its effects on behavior in mice and electroencephalograms in rabbits. AB - We studied the central nervous system (CNS) effects of HSR-609 (3-[4-(8-fluoro 5,11-dihydrobenz[b]oxepino[4,3-b]pyridin-11- ylidene) piperidino]propionic acid dihydrate), a novel amphoteric antiallergic agent having antihistaminic activity. Its effects on the behavior of mice and the electroencephalograms (EEG) of unanesthetized and unrestrained rabbits after oral administration were compared with those of typical antiallergic agents and the non-amphoteric basic compound PY-608 (8-fluoro-5,11-dihydro-11-(1-methyl-4-piperidylidene)benz [b]oxepino[4,3 b]pyridine), which has chemical structure similar to that of HSR-609. HSR-609 (3 300 mg/kg) had no effect on general behavior, spontaneous locomotor activity, hexobarbital-induced sleeping time and reserpine-induced hypothermia in mice. HSR 609 (10-100 mg/kg) and terfenadine (100 mg/kg) had no effect on spontaneous EEG, sleep-wakefulness cycles and EEG power spectra in rabbits. On the other hand, cyproheptadine (3-30 mg/kg), ketotifen (30-100 mg/kg) and PY-608 (0.3-100 mg/kg) caused increases and/or decreases of spontaneous locomotor activity, prolongation of hexobarbital-induced sleeping time and antagonistic effects on reserpine induced hypothermia in mice. These agents (30 mg/kg) increased slow wave sleep and enhanced EEG power spectra at low frequency bands such as delta and theta in rabbits. These findings suggest that HSR-609 has no inhibitory effect on the CNS due to its amphoteric chemical structure. PMID- 9334886 TI - Effects of specific antagonists of angiotensin II receptors and captopril on diabetic nephropathy in mice. AB - We investigated whether angiotension II was involved with diabetic nephropathy in the mouse model. Twelve days after streptozotocin (STZ) injection, the urinary albumin excretion (UAE) level was increased by 118% of the baseline value. On days 21, 28, 35 and 42 after STZ injection, the UAE levels were significantly increased compared with the level at day 12. A marked elevation of creatinine clearance and diabetic-induced renal hypertrophy were also observed on day 49 after STZ injection. The 35-day treatments of captopril and Dup 753 (angiotensin II type 1 receptor antagonist) significantly attenuated the increment of UAE levels (26.4% on day 14 and 34.6% on day 28). PD123177 (angiotensin II type 2 receptor antagonist) also attenuated the increment of UAE (24.7% on day 14) at the dose of 150 mg/kg. Furthermore, Dup 753 partially prevented diabetic-induced renal hypertrophy. These results suggest that angiotensin II type 2 receptor as well as type 1 receptor may be involved in the development of diabetic nephropathy in the STZ-induced diabetic mice, and these mice are beneficial models of early diabetic nephropathy. PMID- 9334887 TI - Repeated antigen inhalation-induced reproducible early and late asthma in guinea pigs. AB - To develop a model of chronic experimental asthma in guinea pigs, the animal was forced to inhale the mist of a low dose of ovalbumin (OA) adsorbed on fine Al(OH)3 for sensitization once every 4 weeks. The animal was challenged by inhalation with the mist of OA on day 14 after the respective sensitizations. Either the first or the second antigen challenge markedly induced an early asthmatic response (EAR), whereas there was hardly any late asthmatic response (LAR). At the 3rd challenge, LAR also emerged with some severity. These dual responses were consistently observed until the 10th challenge. On the other hand, repeated inhalation/challenge, once every 2 weeks, with OA alone at the same dose tended to lead to the desensitization of the EAR. In addition, LAR was hardly observed throughout the experiments. In both groups, gamma 1 and IgE levels in the serum were elevated by the repetitive antigen inhalations, yet no obvious relationship between these antibody levels and the intensity of either EAR or LAR was recognized. The present results indicate that the asthmatic model with reproducible EAR and LAR developed in this study appears to be very beneficial for the investigation of bronchial asthma and for the assessment of anti-asthma drugs. PMID- 9334888 TI - Effects of various selective phosphodiesterase inhibitors on muscle contractility in guinea pig ileal longitudinal smooth muscle. AB - The effects of various selective phosphodiesterase (PDE) inhibitors on muscle contractility in guinea pig ileal longitudinal smooth muscle were investigated. 1) 3-Isobutyl-1-methyl xanthine (IBMX) or zaprinast markedly inhibited the high K(+)- or carbachol (CCh)-induced contraction and increased cGMP content of the muscle strip in a concentration-dependent manner. However, these agents only slightly increased the cAMP content. Milrinone or Ro20-1724 also slightly inhibited the high K(+)- or CCh-induced contraction and increased the cAMP content, but did not increase cGMP. 2) In a fura2-loaded muscle, IBMX or zaprinast inhibited both contractions and the increase in intracellular Ca2+ ([Ca2+]i) level induced by high K+ or CCh, although the inhibitory effect on the [Ca2+]i level was smaller than that on muscle tension. 3) In alpha-toxin permeabilized muscles, cGMP, IBMX or zaprinast significantly inhibited the Ca(2+) induced contraction. These results suggest that IBMX and zaprinast inhibit muscle contraction in the ileal longitudinal smooth muscles mainly through an increase in cGMP and the inhibitory mechanism of IBMX or zaprinast is involved in the decreases in the [Ca2+]i level and sensitivity of contractile elements to Ca2+. PMID- 9334889 TI - Activin selectively abolishes hippocampal long-term potentiation induced by weak tetanic stimulation in vivo. AB - Although modulation of hippocampal synaptic plasticity by neurotrophins or growth factors has recently become an extensively investigated subject, there are no reports arguing for the contribution of the transforming growth factor (TGF)-beta superfamily. In the present study, we examined the effect of activin, a member of the TGF-beta superfamily, on long-term potentiation (LTP) of the dentate gyrus in anesthetized rats. Activin significantly impaired the formation of LTP induced by tetanic stimulation (60 Hz for 0.27 sec), but not that by strong tetanic stimulation (60 Hz for 0.5 sec). These results suggest that activin selectively blocks the induction of LTP evoked by threshold tetanic stimulation. PMID- 9334890 TI - Rolipram, a selective inhibitor of phosphodiesterase type 4, pronouncedly enhanced the forskolin-induced promotion of dopamine biosynthesis in primary cultured rat mesencephalic neurons. AB - A selective inhibitor of cyclic nucleotide phosphodiesterase (PDE) 4, rolipram, markedly enhanced the forskolin-induced increase of intracellular dopamine and dihydroxyphenylacetate (DOPAC, a metabolite of dopamine) levels in primary cultured rat mesencephalic neurons and the forskolin-induced increase of dopamine and DOPAC in extracellular medium. Selective inhibitors of PDE2, PDE3, PDE5 and PDE6 did not have such a promoting effect, and the PDE1 inhibitor vinpocetine and W-7 caused dopamine depletion in the neurons. These findings suggested that PDE4 plays a major role in regulating the intracellular cAMP level to control the dopamine biosynthesis in mesencephalic neurons, whereas PDE1 regulates dopamine release instead. PMID- 9334891 TI - Effect of lecithinized-superoxide dismutase on the interstitial pneumonia model induced by bleomycin in mice. AB - Superoxide anion (O2-) acts as an exacerbation factor in interstitial pneumonia. Lecithinized-superoxide dismutase (PC-SOD), which is synthesized with a lecithin derivative bound covalently to recombinant human Cu,Zn-SOD, has a longer half life in plasma and higher affinity to cell membranes than unmodified SOD. The effect of PC-SOD was evaluated using the bleomycin-induced interstitial pneumonia mouse model. Treatment with PC-SOD at 10 mg/kg significantly reduced the hydroxyproline content and fibrosis score. Namely, PC-SOD suppressed the progression of pulmonary fibrosis on the bleomycin-induced interstitial pneumonia mouse model. PC-SOD may be a potential drug for interstitial pneumonia therapy. PMID- 9334892 TI - Effect of salmon-calcitonin on in vitro opioid withdrawal. AB - The effect on the in vitro withdrawal sign induced by naloxone in morphine dependent guinea pig ileum has been analyzed. Salmon calcitonin (s-CT) dose dependently reduced the force of the contraction induced by naloxone in morphine incubated tissues, but did not modify the contraction induced by administration of acetylcholine or substance P in non-treated tissues. Therefore, the effect of s-CT in morphine incubated tissues may not be attributable to postsynaptic mechanisms, and an inhibitory modulation of the excitatory pathways triggered by naloxone would be suggested. In conclusion, s-CT is able to prevent the withdrawal sign in vitro. PMID- 9334893 TI - Fluoxetine has no marked effect on tic symptoms in patients with Tourette's syndrome: a double-blind placebo-controlled study. AB - To evaluate the efficacy of fluoxetine in the treatment of tics and obsessive compulsive symptoms in patients with Tourette's syndrome (TS), 14 subjects (8-33 years old) with TS participated in a 20-week, fixed-dose (20 mg daily), double blind, placebo-controlled crossover trial of fluoxetine monotherapy. Five subjects met criteria for obsessive-compulsive disorder (OCD), 6 additional subjects had obsessive-compulsive features, and 3 subjects had TS without obsessive-compulsive symptoms. There was no improvement in tics after 8 weeks of treatment with fluoxetine (p = 0.58). In contrast, fluoxetine treatment was associated with a significant reduction in obsessive-compulsive symptoms for the group of 6 subjects initially randomized to fluoxetine (p = 0.04). Crossover analysis showed that fluoxetine had no marked effect on tics (n = 10, p = 0.30, but produced a modest decrease in obsessive-compulsive symptoms (n = 8, p = 0.06). Order effects and carry-over effects were not significant. Withdrawal to placebo was associated with a 55% increase in obsessive-compulsive symptoms (p = 0.05), but there was no effect on tics. The most common side effect was transient behavioral activation, which occurred in about half of the subjects and was more common in children. Fluoxetine may be useful for the treatment of obsessive compulsive symptoms in some patients with TS, but does not appear to be effective for tics. PMID- 9334895 TI - Trazodone is only slightly faster than fluoxetine in relieving insomnia in adolescents with depressive disorders. AB - This retrospective chart review examined the relative effectiveness of fluoxetine and trazodone in relieving insomnia associated with depressive disorders in adolescents (aged 13-17 years). We reviewed the hospital charts of consecutively admitted adolescents with a depressive disorder and insomnia, who received one of three treatments: fluoxetine (20 +/- 2.2 mg), trazodone (71 +/- 32 mg), or a fluoxetine-trazodone combination (fluoxetine 29 +/- 2.2 mg, trazodone 68 +/- 29 mg). Each treatment was examined in 20 patients. Insomnia was defined as a change in sleep patterns characterized by decreased total sleep time that was sufficient to cause clinical concern, and insomnia resolution was defined as sleep starting by midnight and lasting 6 hours. Mean time to resolution of insomnia was significantly faster in adolescents treated with trazodone rather than fluoxetine (2.5 vs. 5.1 days, p < 0.05). Trazodone seemed to save only about 3 days and insomnia resolved in all subjects by the 11th day of antidepressant treatment. Median time to insomnia resolution was 2 days (range 1-5 days) in the trazodone group and 4 days (range 1-11 days) in the fluoxetine group. This difference between trazodone and fluoxetine, although statistically significant, was generally not clinically significant in the management of insomnia associated with depressive disorders in adolescents. The resolution of insomnia was not faster for treatment with a combination of fluoxetine and trazodone in comparison to fluoxetine monotherapy. Insomnia resolution was slightly later in older children. These clinical findings await confirmation by a controlled study. Both drugs seemed effective in ameliorating sleep symptoms in this sample, although it is likely that they produced these changes by different mechanisms. PMID- 9334894 TI - Fluoxetine in drug-dependent delinquents with major depression: an open trial. AB - Although fluoxetine might be more effective than placebo for treating adolescent depression without major comorbidity, little is known about the response of depressive symptoms to antidepressants in adolescents with comorbid conduct disorder (CD) and substance use disorders (SUD). Male adolescents, who remained or became depressed after > or = 1 month of abstinence from abused substances during residential treatment for SUD, were treated in an open trial for > or = 7 weeks with a fixed dose of 20 mg of fluoxetine. The eight adolescents (ages 14-18 years) with CD, SUD, and major depression were not in drug withdrawal or receiving other pharmacotherapy. A > or = 50% improvement was observed in mean scores on Ten Point Depression Scale rated by clinician (p < 0.01) and patients (p < 0.01), Carroll Self-Ratings for depression (p < 0.02), and Severity of Illness scores on the Clinical Global Impression (p < 0.01). Of the eight adolescents, seven showed marked improvement and wished to continue fluoxetine after the trial. Side effects were mild and transient. No subject required dosage reduction or discontinuation of medication because of side effects. Fluoxetine appeared useful in treating substance-dependent delinquents whose major depressions persisted or emerged after 4 weeks of abstinence. These preliminary findings justify a controlled trial in such youths. PMID- 9334896 TI - Clomipramine in adults with pervasive developmental disorders: a prospective open label investigation. AB - The purpose of this investigation was to determine the short-term efficacy and tolerability of clomipramine in a consecutive series of adults with pervasive developmental disorders (PDDs). Thirty-five adults with PDDs (DSM-IV), 16 of whom were nonverbal, entered a 12-week prospective open-label trial of clomipramine. The initial sample included 18 patients with autistic disorder, 6 patients with Asperger's disorder, and 11 patients with pervasive developmental disorder not otherwise specified (PDDNOS). Behavioral ratings were obtained at baseline and after 4, 8, and 12 weeks of clomipramine. Eighteen (55%) of the 33 patients who completed the trial were categorized as treatment responders based on scores of "much improved" or "very much improved" on the Clinical Global Impression (CGI) global improvement item (p < 0.001). Ten (63%) of 16 patients with autistic disorder, 2 (33%) of 6 patients with Asperger's disorder, and 6 (55%) of 11 patients with PDDNOS were considered responders to clomipramine treatment. In those 18 patients, clomipramine significantly reduced total repetitive thoughts and behavior (p < 0.001) and also aggression (p < 0.001), and improved some aspects of social relatedness, such as eye contact and verbal responsiveness (p < 0.001). Change in these specific symptom clusters over time was not related to DSM-IV subtype of PDD. The level of autistic behavior, as measured by the Autism Behavior Checklist (ABC) score, and full-scale intelligence quotient (IQ) were not significantly associated with global treatment response. Whereas clomipramine was well tolerated by most patients, 13 had clinically significant adverse effects. Three patients had seizures during clomipramine treatment, including 2 who had prior seizure disorders and were taking anticonvulsants. Of the 32 patients who had no history of prior seizures, only 1 had a seizure during clomipramine treatment. There were no adverse cardiovascular or extrapyramidal effects. All responders continued on clomipramine after completion of the study. The results of this open-label trial suggest that clomipramine may be an effective drug for reducing repetitive thoughts and actions and aggressive behavior and for improving some elements of social behavior, such as eye contact and verbal responsivity in adults with PDDs. Careful monitoring of adverse effects, particularly seizures, is warranted. Although an electroencephalogram (EEG) is not mandatory in patients with PDD prior to clomipramine treatment, we recommend that patients with PDD and a history of seizures be treated initially with a selective serotonin uptake inhibitor rather than with clomipramine. The findings of this study require replication in a double-blind placebo-controlled investigation before definitive statements of efficacy and tolerability can be made. PMID- 9334897 TI - Effects of methylphenidate and behavioral contingencies on sustained attention in attention-deficit hyperactivity disorder: a test of the reward dysfunction hypothesis. AB - Psychostimulants and behavior therapy have been postulated to be effective in treating attention-deficit hyperactivity disorder (ADHD) by compensating for a pathologically elevated reward threshold, but no studies have compared reinforcement to psychostimulants in maintaining task performance. The separate and combined effects of methylphenidate (MPH, 0.6 mg/kg) and a behavioral intervention (reward plus response cost) were assessed on a continuous performance test (CPT, a measure of sustained attention) modified to deliver auditory feedback contingent upon the subject's responses. Each of 22 children (6 10 years old) with ADHD were tested under four treatment conditions: placebo + feedback, placebo + behavioral contingencies, MPH + feedback, and MPH + contingencies. CPT performance, indexed by d' (ability to discriminate between target and false targets), was significantly better with MPH than with placebo, showing reduced deterioration over time. Contingency treatment improved mean d' compared to placebo + feedback but, in contrast, had no effect on the slope of performance deterioration. Addition of contingencies to MPH did not yield further improvement. The results indicate that MPH improved sustained attention on a laboratory task (and reduced task-irrelevant and other disinhibited behaviors), whereas behavioral contingencies did not. These findings suggest that, although both interventions improved stimulus discrimination processes, only MPH enhanced processes that mediate the regulation of effort over time. In addition, the disjunction between the effects of reward and of MPH provides evidence that psychostimulant effects on attention are only partially explained by the stimulation of brain centers associated with reward. PMID- 9334898 TI - Buspirone treatment of anxiety associated with pharyngeal dysphagia in a four year-old. AB - Buspirone is a nonbenzodiazepine anxiolytic that has been effective in uncontrolled trials for treating childhood anxiety disorders. A 4-year-old boy with a history of laryngomalacia (congenital structural abnormality with airway collapse and obstruction on inhalation), pharyngeal dysphagia (difficulty in swallowing), poor weight gain, delayed self-feeding skills, and anxiety symptoms is described. An open trial of buspirone, increased gradually to 12.5 mg daily in divided doses over a period of 22 weeks, was associated with decreased anxiety, improved self-feeding skills, and weight gain. Based on parental reports, buspirone appeared to decrease separation and social anxiety, as well as anxiety associated with eating. Drug discontinuation was associated with symptom relapse, whereas drug readministration lead to the same clinical benefits that had been observed previously. The medication was well tolerated, and its benefits have persisted for over 1 year. No new recommendations can be made regarding the use of buspirone in preschool children or in the treatment of anxious behaviors adversely affecting medical conditions in children and adolescents. PMID- 9334899 TI - Modelling of the binding site of the human m1 muscarinic receptor: experimental validation and refinement. AB - Our model of the human m1 muscarinic receptor has been refined on the basis of the recently published projection map of bovine rhodopsin. The refined model has a slightly different helix arrangement, which reveals the presence of an extra hydrophobic pocket located between helices 3, 4 and 5. The interaction of series of agonists and antagonists with the m1 muscarinic receptor has been studied experimentally by site-directed mutagenesis. In order to account for the observed results, three-dimensional models of m1 ligands docked in the target receptor are proposed. Qualitatively, the obtained models are in good agreement with the experimental observations. Agonists and partial agonists have a relatively small size. They can bind to the same region of the receptor using, however, different anchoring receptor residues. Antagonists are usually larger molecules, filling almost completely the same pocket as agonists. They can usually produce much stronger interactions with aromatic residues. Experimental data combined with molecular modelling studies highlight how subtle and diverse receptor-ligand interactions could be. PMID- 9334900 TI - QXP: powerful, rapid computer algorithms for structure-based drug design. AB - New methods for docking, template fitting and building pseudo-receptors are described. Full conformational searches are carried out for flexible cyclic and acyclic molecules. QXP (quick explore) search algorithms are derived from the method of Monte Carlo perturbation with energy minimization in Cartesian space. An additional fast search step is introduced between the initial perturbation and energy minimization. The fast search produces approximate low-energy structures, which are likely to minimize to a low energy. For template fitting, QXP uses a superposition force field which automatically assigns short-range attractive forces to similar atoms in different molecules. The docking algorithms were evaluated using X-ray data for 12 protein-ligand complexes. The ligands had up to 24 rotatable bonds and ranged from highly polar to mostly nonpolar. Docking searches of the randomly disordered ligands gave rms differences between the lowest energy docked structure and the energy-minimized X-ray structure, of less than 0.76 A for 10 of the ligands. For all the ligands, the rms difference between the energy-minimized X-ray structure and the closest docked structure was less than 0.4 A, when parts of one of the molecules which are in the solvent were excluded from the rms calculation. Template fitting was tested using four ACE inhibitors. Three ACE templates have been previously published. A single run using QXP generated a series of templates which contained examples of each of the three. A pseudo-receptor, complementary to an ACE template, was built out of small molecules, such as pyrrole, cyclopentanone and propane. When individually energy minimized in the pseudo-receptor, each of the four ACE inhibitors moved with an rms of less than 0.25 A. After random perturbation, the inhibitors were docked into the pseudo-receptor. Each lowest energy docked structure matched the energy-minimized geometry with an rms of less than 0.08 A. Thus, the pseudo receptor shows steric and chemical complementarity to all four molecules. The QXP program is reliable, easy to use and sufficiently rapid for routine application in structure-based drug design. PMID- 9334901 TI - Electron affinities of p-benzoquinone, p-benzoquinone imine and p-benzoquinone diimine, and spin densities of their p-benzosemiquinones computed by several quantum chemical models. AB - Restricted and unrestricted (U) Hartree-Fock (HF), second-order Moller-Plesset perturbation (MP2), density functional (DF), hybrid HF/DF and semiempirical (half electron (HE) method) models have been used to calculate adiabatic electron affinities (EAad values) of p-benzoquinone (I), p-benzoquinone imine (VI) and p benzoquinone diimine (XI), as well as expectation values () and spin density distributions in the radical anions of I, VI and XI. The AM1/AM1-HE and ab initio calculated structures are found to be in accord with each other. The ROHF/6 31G(d) method gave the poorest EAad result. The UHF and UMP2 wave functions were found to be substantially spin contaminated (for the radicals) and the accuracies of the EAad values calculated were also poor. The use of molecular energies obtained after spin annihilation did not lead to significant improvement of the UHF and UMP2 results. In contrast to the ROHF, UHF and UMP2 results, the DF(USVWN, UBVWN, UBLYP) and hybrid HF/DF(UB3LYP) methods, as well as the AM1-HE, gave much better results. The calculated EAad values decreased, as predicted by most of the models, in the order EAad(I) > EAad(VI) > EAad(XI). The differences in the EAs, EAad(I)-EAad(VI) and EAad(I)-EAad(XI), were consistently predicted to be about 8-9 and 17-18 kcal/mol, respectively, by the DF, B3LYP and AM1-HE models. The performance of the PM3 and SAM1 models was not as good as the AM1 model. Of all the methods tested, the B3LYP/6-311G(d,p) model is concluded to give the most accurate quantitative trend (I(42.6) > VI(33.1) > XI(23.7)) in EAad. The predicted trend in EA can satisfactorily be rationalized by the calculated LUMO orbital energies, atomic charges and spin density distributions. Analysis of the spin density data predicts that phenoxyl- and anilino-type radical anions predominate in the p-benzosemiquinones of I and XI, respectively, while both phenoxyl- and anilino-type radicals contribute to the structure of the p-benzosemiquinone of VI, with the anilino-type predominating. PMID- 9334902 TI - Time-efficient flexible superposition of medium-sized molecules. AB - We present an efficient algorithm for the structural alignment of medium-sized organic molecules. The algorithm has been developed for applications in 3D QSAR and in receptor modeling. The method assumes one of the molecules, the reference ligand, to be presented in the conformation that it adopts inside the receptor pocket. The second molecule, the test ligand, is considered to be flexible, and is assumed to be given in an arbitrary low-energy conformation. Ligand flexibility is modeled by decomposing the test ligand into molecular fragments, such that ring systems are completely contained in a single fragment. Conformations of fragments and torsional angles of single bonds are taken from a small finite set, which depends on the fragment and bond, respectively. The algorithm superimposes a distinguished base fragment of the test ligand onto a suitable region of the reference ligand and then attaches the remaining fragments of the test ligand in a step-by-step fashion. During this process, a scoring function is optimized that encompasses bonding terms and terms accounting for steric overlap as well as for similarity of chemical properties of both ligands. The algorithm has been implemented in the FLEXS system. To validate the quality of the produced results, we have selected a number of examples for which the mutual superposition of two ligands is experimentally given by the comparison of the binding geometries known from the crystal structures of their corresponding protein-ligand complexes. On more than two-thirds of the test examples the algorithm produces rms deviations of the predicted versus the observed conformation of the test ligand below 1.5 A. The run time of the algorithm on a single problem instance is a few minutes on a common-day workstation. The overall goal of this research is to drastically reduce run times, while limiting the inaccuracies of the model and the computation to a tolerable level. PMID- 9334903 TI - Multiple automatic base selection: protein-ligand docking based on incremental construction without manual intervention. AB - A possible way of tackling the molecular docking problem arising in computer aided drug design is the use of the incremental construction method. This method consists of three steps: the selection of a part of a molecule, a so-called base fragment, the placement of the base fragment into the active site of a protein, and the subsequent reconstruction of the complete drug molecule. Assuming that a part of a drug molecule is known, which is specific enough to be a good base fragment, the method is proven to be successful for a large set of docking examples. In addition, it leads to the fastest algorithms for flexible docking published so far. In most real-world applications of docking, large sets of ligands have to be tested for affinity to a given protein. Thus, manual selection of a base fragment is not practical. On the other hand, the selection of a base fragment is critical in that only few selections lead to a low-energy structure. We overcome this limitation by selecting a representative set of base fragments instead of a single one. In this paper, we present a set of rules and algorithms to automate this selection. In addition, we extend the incremental construction method to deal with multiple fragmentations of the drug molecule. Our results show that with multiple automated base selection, the quality of the docking predictions is almost as good as with one manually preselected base fragment. In addition, the set of solutions is more diverse and alternative binding modes with low scores are found. Although the run time of the overall algorithm increases, the method remains fast enough to search through large ligand data sets. PMID- 9334904 TI - Conformational analysis of six- and twelve-membered ring compounds by molecular dynamics. AB - A molecular dynamics (MD)-based conformational analysis has been performed on a number of cycloalkanes in order to demonstrate the reliability and generality of MD as a tool for conformational analysis. MD simulations on cyclohexane and a series of methyl-substituted cyclohexanes were performed at temperatures between 400 and 1200 K. Depending on the simulation temperature, different types of interconversions (twist-boat-twist-boat, twist-boat-chair and chair-chair) could be observed, and the MD simulations demonstrated the expected correlation between simulation temperature and ring inversion barriers. A series of methyl substituted 1,3-dioxanes were investigated at 1000 K, and the number of chair chair interconversions could be quantitatively correlated to the experimentally determined ring inversion barrier. Similarly, the distribution of sampled minimum energy conformations correlated with the energy-derived Boltzmann distribution. The macrocyclic ring system cyclododecane was subjected to an MD simulation at 1000 K and 71 different conformations could be sampled. These conformations were compared with the results of previously reported conformational analyses using stochastic search methods, and the MD method provided 19 out of the 20 most stable conformations found in the MM2 force field. Finally, the general performance of the MD method for conformational analysis is discussed. PMID- 9334905 TI - Three-dimensional modelling of human cytochrome P450 1A2 and its interaction with caffeine and MeIQ. AB - The three-dimensional modelling of proteins is a useful tool to fill the gap between the number of sequenced proteins and the number of experimentally known 3D structures. However, when the degree of homology between the protein and the available 3D templates is low, model building becomes a difficult task and the reliability of the results depends critically on the correctness of the sequence alignment. For this reason, we have undertaken the modelling of human cytochrome P450 1A2 starting by a careful analysis of several sequence alignment strategies (multiple sequence alignments and the TOPITS threading technique). The best results were obtained using TOPITS followed by a manual refinement to avoid unlikely gaps. Because TOPITS uses secondary structure predictions, several methods that are available for this purpose (Levin, Gibrat, DPM, NnPredict, PHD, SOPM and NNSP) have also been evaluated on cytochromes P450 with known 3D structures. More reliable predictions on alpha-helices have been obtained with PHD, which is the method implemented in TOPITS. Thus, a 3D model for human cytochrome P450 1A2 has been built using the known crystal coordinates of P450 BM3 as the template. The model was refined using molecular mechanics computations. The model obtained shows a consistent location of the substrate recognition segments previously postulated for the CYP2 family members. The interaction of caffeine and a carcinogenic aromatic amine (MeIQ), which are characteristic P450 1A2 substrates, has been investigated. The substrates were solvated taking into account their molecular electrostatic potential distributions. The docking of the solvated substrates in the active site of the model was explored with the AUTODOCK programme, followed by molecular mechanics optimisation of the most interesting complexes. Stable complexes were obtained that could explain the oxidation of the considered substrates by cytochrome P450 1A2 and could offer an insight into the role played by water molecules. PMID- 9334906 TI - Evaluation of a novel infrared range vibration-based descriptor (EVA) for QSAR studies. 1. General application. AB - A novel molecular descriptor (EVA) based upon calculated infrared range vibrational frequencies is evaluated for use in QSAR studies. The descriptor is invariant to both translation and rotation of the structures concerned. The method was applied to 11 QSAR datasets exhibiting both a range of biological endpoints and various degrees of structural diversity. This study demonstrates that robust QSAR models can be obtained using the EVA descriptor and examines the effect of EVA parameter changes on these models; recommendations are made as to the appropriate choice of parameters. The performance of EVA was found to be comparable in statistical terms to that of CoMFA, despite the fact that EVA does not require the generation of a structural alignment. Models derived using semiempirical (MOPAC AM1 and PM3) and AMBER mechanics calculated normal mode frequencies are compared, with the overall conclusion that the semiempirical methods perform equally well and both outperform the AMBER-based models. PMID- 9334907 TI - 24-hour ambulatory blood pressure and renal disease in young subjects with type I diabetes. AB - Twenty-four hour ambulatory blood pressure (ABP) was evaluated in 150 teenage and young adults with insulin-dependent diabetes mellitus (IDDM) to define high-risk subjects who are likely to develop early diabetic nephropathy. Their age range was 16-28 years with diabetes of 3.5-25.8 years duration. All subjects had office blood pressure (BP) measured, wore an ABP monitor for 24 h, and collected two or more timed urine samples for albumin excretion rates (AERs). Eighty-six subjects had no elevation of AER (< 7.6 micrograms/min), 29 subjects had borderline elevations (7.6-20 micrograms/min), 24 subjects had microalbuminuria (20.1-200 micrograms/min), and 11 had macroalbuminuria (> 200 micrograms/min). Age, gender, duration of diabetes, and single office BP readings were similar in the four groups (p > 0.05, logistic regression). All 24-h ABP parameters were significantly higher in subjects with diabetes having AER values greater than 7.6 micrograms/min when compared with healthy age- and gender-matched nondiabetic controls (p < 0.05, Dunnett's t test). The percent of nighttime systolic and diastolic ABP readings above the 90th percentile of normal for age, gender, and race and the percent of readings in the hypertensive range (> 140/90) were significantly related with AERs (p < 0.01; logistic regression). A higher percentage of ABP values above the 90th percentile for age, gender, and ethnic group or of ABP readings above hypertensive levels (> or = 140/90) are associated with diabetic renal disease. PMID- 9334908 TI - The impact of coexistent diabetes on the prevalence of coronary heart disease. AB - The increased risk of developing cardiovascular disease in diabetic population has been well documented, but the prevalent mechanism of this susceptibility is still only partly explained. We compared the impact of diabetes on ischemic heart disease in patients hospitalized in a public general hospital over a 10-year period. The prevalence of coronary heart disease (CHD) was consistently higher among diabetic population [namely, among non-insulin-dependent diabetes mellitus (NIDDM) patients] when compared with the nondiabetic population. The prevalence was similar in both genders, increasing with age, and was independent from body mass index, history of smoking, metabolic control, or lipid pattern. Heart rate and blood pressure levels were significantly higher in NIDDM patients with CHD; similarly, there was a significant association between ischemic heart disease and atherosclerotic peripheral artery disease prevalence, and this trend was observed even in subjects with impaired glucose tolerance. These observations support the evidence that diabetes exerts a deleterious effect on general risk factors of atherosclerosis and increases susceptibility to cardiovascular disease by itself as an "independent" risk factor; on the other hand, the epidemiological evidence of an excessive occurrence of type II diabetes in individuals with pre-existing vascular disease suggests a genetically determined link between metabolic disturbances and cardiovascular disease. PMID- 9334909 TI - The effect of pentoxifylline on current perception thresholds in patients with diabetic sensory neuropathy. AB - Diabetic peripheral neuropathy is a common, painful, and disabling condition that is known to occur by two mechanisms: hyperglycemia and arterial blood flow occlusion. Pentoxifylline (Trental) functions by improving erythrocyte flexibility in blood vessels, which could increase the delivery of oxygen to occluded nerve vessels. This 1-year clinical trial was aimed at ascertaining the effects of pentoxifylline on diabetic sensory neuropathy. Fifty patients with type I or II diabetes were evaluated in a randomized, double-blind, parallel group and placebo-controlled study. Pentoxifylline effectiveness was evaluated by measuring glycated hemoglobin, blood pressure and current perception threshold (CPT). The CPT results showed no statistically significant effect of pentoxifylline on mean nerve sensory perception thresholds in ankle and toe at 5, 250 and 2000 Hz. There were no significant changes in glycated hemoglobin or in systolic and diastolic blood pressure during the trial. Thus, glycated hemoglobin and blood pressure did not explain the lack of pentoxifylline effect on diabetic neuropathy. In conclusion, pentoxifylline appears not to add benefits to the clinical treatment of diabetic sensory neuropathy of the lower extremity. PMID- 9334910 TI - Renal arterial reactivity to potassium, noradrenaline, and neuropeptide Y and association with urinary albumin excretion in the diabetic rat. AB - A changed vasomotor reactivity of renal arteries may lead to defect autoregulation of renal hemodynamics with damage of diabetic kidneys. Eleven streptozotocin-induced diabetic male Wistar rats were daily treated with insulin in order to achieve a blood glucose of 21 mmol/L. Seventeen age and gender matched rats served as controls. After 50 days, the kidneys were rapidly removed, arteria renalis and the first branches of the intrarenal arteries were dissected free. The arterial reactivity was tested with a sensitive in vitro method. The reactivity to noradrenaline was tested by cumulative application (10(-9) to 3 x 10(-4) M) before and after a single concentration of neuropeptide Y (NPY). Potassium (60 mM) and noradrenaline induced a strong contraction of all arteries with similar response in diabetic and control rats. The effect of noradrenaline after NPY was unchanged in renal vessels of control rats, whereas it was diminished in intrarenal vessels for both diabetic and control rats. Similarly, a diminished response was found for renal arteries in diabetic rats, an effect which was related to the level of blood glucose (r = 0.62, 2p = 0.04). The urinary excretion rate of albumin in the diabetic rats was related to the largest noradrenaline induced contraction (r = 0.71, 2p = 0.01) of renal but not of intrarenal arteries. In conclusion, there was no difference in potassium and noradrenaline evoked contractions in renal and intrarenal arteries in diabetic and control rats. NPY decreased the contractile response to noradrenaline. The high blood glucose slightly increased this effect of NPY. PMID- 9334911 TI - Outcome of trigger finger treatment in diabetes. AB - Trigger finger is an underdiagnosed hand disorder causing disability in longstanding diabetic patients. Sixty diabetic patients [39 insulin-dependent diabetes mellitus (IDDM) and 21 non-insulin-dependent diabetes mellitus (NIDDM)] and 60 nondiabetic patients were examined. All were initially treated by steroid injections: failure to alleviate symptoms was the indication for surgery. The incidence of multiple digit involvement was higher in IDDM patients as compared with the control group (p < 0.001). The diffuse type was 1.45 times more frequent in IDDM and NIDDM than in nondiabetic patients (p < 0.008). The diabetic patients had a relatively longer duration of symptoms (p < 0.003). Significantly, a higher recovery rate upon steroid injection was achieved in control patients as compared with the diabetic ones (p < 0.001). IDDM patients required more surgery compared with NIDDMs and, in 13.3% of diabetic patients, the surgical outcome was not successful. Diabetic patients should be diagnosed early for multiple and diffuse types of trigger digits. Steroid injection as the first mode of therapy is highly recommended although not always successful. Surgery is the definitive treatment but requires a long course of physiotherapy and may be associated with some complications. PMID- 9334912 TI - Comparison of the microvascular response to transcutaneous electrical nerve stimulation and postocclusive ischemia in the diabetic foot. AB - Neurogenic inflammation, mediated by unmyelinated C-nerve fibers, is part of the acute neurovascular response to injury. Laser doppler flowmetry was used to measure the flare response to transcutaneous electrical nerve stimulation (TENS) and to compare this axon reflex vasodilatation with postischemic hyperemia in the skin of the foot in diabetic and nondiabetic subjects. Twenty-one control subjects and 57 diabetic patients (25 type I; 32 type II; 14 without complications; 14 with neuropathy and without retinopathy; 8 with retinopathy and without neuropathy; 21 with neuropathy and retinopathy) were enrolled in the study. Following TENS, an increase in skin blood flow was found at the dorsum of the foot without any significant difference between the different groups. Compared to the control group, axon reflex vasodilatation was significantly reduced in the group of patients with diabetic neuropathy and in the group of patients with diabetic neuropathy and retinopathy at the pulp of the hallux (61 +/- 15 versus -6 +/- 16; versus 23 +/- 5; p < 0.05, respectively). All investigated groups exhibited a significant increase in skin blood flow after arterial occlusion without any significant difference between the groups. A good association was observed between postocclusive and TENS-induced hyperemia at the dorsum of the foot (r = 0.39; p = 0.0002), but only a weak association was found at the pulp of the hallux (r = 0.24; p = 0.03). TENS-induced hyperemia was associated with a diminished sweat response (p = 0.03), but not with pathological cardiovascular function tests (p = 0.07). Electrical axon reflex vasodilatation is diminished in diabetic patients suffering from peripheral autonomic C-fiber injury, especially in skin rich in thermoregulatory blood flow. The diminished neurovascular response is independent of vascular alterations in diabetes mellitus. PMID- 9334914 TI - Community: meaning and opportunity, and learning for the future. PMID- 9334913 TI - Three-year follow-up on scintigraphically assessed cardiac sympathetic denervation in patients with long-term insulin-dependent (type I) diabetes mellitus. AB - Scintigraphy using I-123-metaiodobenzylguanidine (I-123-MIBG) and Tc-99m methoxyisobutylisonitrile (Tc-99m-MIBI) allows assessment of the cardiac sympathetic innervation and the myocardial perfusion. To investigate the natural history of cardiac sympathetic denervation in long-term diabetic patients without myocardial perfusion defects, global and regional I-123-MIBG and Tc-99m-MIBI uptake was determined (score 1-6; 1 = normal uptake, 6 = no uptake) in 22 patients with insulin-dependent (type I) diabetes mellitus (IDDM) at 3-year follow-up. All patients were treated with intensive insulin therapy and HbA1c was 8.0% +/- 1.0% at entry compared with 7.9% +/- 1.1% at follow-up. Cardiac sympathetic denervation (I-123-MIBG uptake score > 2), initially observed in 18 patients, was detectable in 21 patients at follow-up. The global myocardial I-123 MIBG uptake score deteriorated in eight patients, remained unchanged in 11 and improved in three patients. The changes in mean global I-123-MIBG uptake score (3.5 +/- 1.0 versus 3.8 +/- 0.8) were not significant. Reduction of the anterior, lateral, posterior, septal, and apical I-123-MIBG uptake did not progress significantly during follow-up. The mean uptake score of the posterior myocardial region (4.7 +/- 0.8) was smaller than the uptake score of the anterior (3.0 +/- 1.1, p = 0.001), lateral (3.2 +/- 0.9, p < 0.001) and septal (4.1 +/- 1.1, p < 0.05) myocardial regions. At follow-up, moderate myocardial perfusion defects (global Tc-99m-MIBI uptake score = 3) were detectable in four patients. Our study demonstrates that scintigraphically assessed cardiac sympathetic denervation does neither significantly regress nor progress on the average in a group of long-term IDDM patients during a 3-year follow-up. Thus, it is concluded that cardiac sympathetic abnormalities are a persistent, yet frequent phenomenon in long-term IDDM patients. PMID- 9334915 TI - Paradigms lost: the persisting search for community in U.S. health policy. AB - Local communities have long played an important role in health and social policy in the United States. But the concept of community was strangely absent from the federal debate on health care reform in 1993 and 1994. I attribute this absence to the paradoxical nature of community as a frame for guiding policy making. The concept of community has broad appeal across the ideological spectrum, but this breadth masks a set of long-standing and powerful tensions that determine when communities are seen as appropriately given responsibility for addressing societal problems. This article reviews the historical evolution of the role of community in health policy, highlighting the ways in which various tensions emerged. It applies these perspectives to an analysis of the attitudes of the U.S. public and congressional staff in 1995. Data from two surveys demonstrate that support for community-based reforms is much lower for medical care than for other social needs, including some health-related concerns such as long-term care and the treatment of substance abuse. Statistical analyses suggest several possible explanations for the lower support for community-based medical care. The article concludes with a discussion of the implications for future communitarian strategies designed to improve U.S. medicine or social policy. PMID- 9334916 TI - Enemies of the people: the moral dimension to public health. AB - This essay explores the effects of morality on health policy. Moral images and stereotypes, I argue, have powerful political consequences. They are the differences between fighting poverty and fearing the poor, between expanding social welfare programs and cracking down on crime, between public health campaigns and drug wars. I begin by locating morality within traditional paradigms of American politics (which are designed to overlook the issue); I then suggest how moral stigmas are constructed; show how they are deployed in debates over public health issues, such as alcohol abuse and drug addiction; and briefly sketch an alternative approach to defining community and seeking public health. PMID- 9334917 TI - Community control in a world of regional delivery systems. AB - The pell-mell restructuring of health care into massive regional delivery systems has disrupted long-standing relationships between local leaders and residents and their community health care systems. This diminished role of communities in our new world of health care is ironic. As control within large regions in this country becomes concentrated within the operation of three or four health plans, we become increasingly dependent upon oligopolies for our market solutions. As economic arrangements, all that oligopolies can offer are indeterminate outcomes. Some may be good for consumers, others disastrous. Without the countervailing influence of nonmarket community interests, individuals may find their satisfaction with the health care system greatly diminished. PMID- 9334918 TI - Community experiments in action: developing community-defined models for reconfiguring health care delivery. AB - In the United States, health system change occurs as the interaction of politics and policies is played out and pushed forward by individuals and organizations taking action in their local communities and markets. This article provides context and descriptive information about a model of local health care reform that is being tested in twenty-five communities around the nation. We introduce a value-based model of community-responsive health system change, the Community Care Network (CCN) Vision. We briefly describe a national demonstration program that is testing this model between January 1996 and the end of 1998. We offer several ways of looking at the local and regional multisectoral partnerships that are attempting to demonstrate the CCN vision. We look at their composition, the kinds of actions in which they are engaged, several example challenges and responses to them, and we give several examples of schools becoming part of the health system of last resort. Finally, we present some early ideas on how the local actions described here may influence (and be influenced by) the political and policy contexts in which they occur. PMID- 9334919 TI - The limits of social learning: translating analysis into action. AB - In what respects does public-policy making reflect social learning, drawing lessons from previous experiences and from the experiences of governments in other settings? Starting with an examination of the effect of policy legacies on current policy making, I present a process model of social learning embedded within the larger policy-making process resting at the intersection of the nation's constitutional context, technological change, and political influences exogenous to social learning. The model first distinguishes between the structural and the social learning effects of policy legacies. I then conceptually divide social learning into separate streams of substantive learning and situational learning. The effect that each of these has on policy making depends on the relative position of three categories of participants in the policy-making process (experts, organized interests, and politicians), as well as on the scope of the policy issue being considered (ranging from routine change to major reform). This analysis, with reference to recent health care policy making, reveals the full extent to which social learning is often a decidedly political struggle over ideas and information in which advocates promote lessons that severe their specific interests within a given institutional context and political setting. I consider the implications of social learning for understanding likely policy responses to the rise of market forces in health care. PMID- 9334920 TI - Infection of a human retinal pigment epithelial cell line with human herpesvirus 6 variant A. AB - A retinal pigment epithelial (RPE) cell line (K-1034) was examined for its susceptibility to human herpesvirus 6 variant A (HHV-6A). Exposure of K-1034 cells to HHV-6A induced the formation of multinucleated giant cells, which was suppressed by an inhibitor of viral DNA synthesis. In the giant cells, herpesvirus nucleocapsids were demonstrated by electron microscopy and the viral glycoprotein B was detected by immunofluorescence assay. These results indicate that K-1034 cells are susceptible to HHV-6A and suggest that HHV-6A has an ability to directly destroy epithelial cells. PMID- 9334921 TI - Relationship between human papillomavirus infection and overexpression of p53 protein in cervical carcinomas and lymph node metastases. AB - Overexpression of p53 protein is common in cervical carcinoma. We investigated archival biopsies from 26 cervical cancer patients (24 with available lymph nodes) to determine the relationship between p53 overexpression and HPV infection at the cervix and lymph nodes. Twelve cervical carcinoma patients had p53 protein in cervical biopsies detectable by immunohistochemistry using monoclonal antibody DO-1, and 22 were positive for HPV DNA in polymerase chain reaction assays (16, contained HPV-16; 3, HPV-18; and, 3 HPV-X). Seven cervical cancer patients had one or more lymph nodes positive for p53 protein, and all but one of these were concordantly p53 positive at the cervix. However, detection of p53 protein in cervical biopsies was predictive neither of the expression of p53 at draining lymph nodes (P > 0.1) nor of the occurrence of metastases (P > 0.1). Fourteen patients were positive at one or more lymph nodes for HPV DNA. Cervical positivity for HPV DNA was associated significantly with concordant HPV positivity at the lymph nodes (P = 0.039) and was predictive of metastases (P = 0.019). There was no association between positivity for p53 and for HPV DNA at primary cervical carcinomas or at the lymph nodes (all P > 0.1). We conclude that, although detectable p53 protein is a common feature of cervical carcinomas, it is not predictive of metastases and is independent of HPV infection. PMID- 9334922 TI - Mutations in the NS5A gene of hepatitis C virus in North American patients infected with HCV genotype 1a or 1b. AB - Previous studies from Japan have described an association between a conserved sequence within the hepatitis C virus (HCV) genome and resistance to interferon (IFN) therapy for patients infected with HCV genotype 1b [Enomoto et al. (1995): Journal of Clinical Investigation 96: 224-230; Enomoto et al. (1996): New England Journal of Medicine 334:77-81]. The present study examines amino acid sequences surrounding the putative Interferon Sensitivity Determining Region (ISDR) of the NS5A gene of HCV in 21 North American patients with genotype 1a or 1b infection receiving recombinant IFN therapy. The ISDR consensus or intermediate pattern was observed in 13 of 14 NS5A clones from North American patients infected with genotype 1b. However, we found no evidence of the consensus ISDR sequence in any NS5A clones isolated from 15 patients with genotype 1a infection. In select cases, gel shift analysis showed no significant changes in the clonal frequency of the putative ISDR domain of HCV genotype 1a or 1b infected patients who were either nonresponsive to IFN therapy, or relapsed following withdrawal of IFN therapy. These results suggest that a conserved ISDR domain is neither associated with, nor responsible for, IFN resistance in North American patients infected with HCV genotype 1a, and demonstrate a need for further investigation into the reported association between ISDR consensus sequences and IFN resistance in genotype 1b clones. PMID- 9334923 TI - Differentiation of core gene products of the hepatitis B virus in infected liver tissue using monoclonal antibodies. AB - Hepatitis B virus (HBV) core gene translational products were localised previously in the cytoplasm and/or in the nuclei of infected cells. We investigated in naturally infected human hepatocytes whether this variation in the subcellular expression is due to differences in the presence of assembled core particles and other core gene derived proteins, the expression of HBeAg and the processing of liver tissue. By immunostaining of liver specimens infected with HBeAg-positive and HBeAg-minus variants of HBV, using monoclonal antibodies specific for assembled core particles and for various epitopes on denatured core protein, it was shown that virtually all immunoreactive core gene products are assembled into core particles. The latter are present both in the nuclei and in the cytoplasm of hepatocytes, independent of the infecting virus strain. A marked reduction or absence of immunoreactivity, observed with some monoclonal antibodies, was shown to result from nucleotide sequence variations within or close to the corresponding epitope. These results demonstrate that immunoreactive products, derived from the HBV core gene, in the nuclei and cytoplasm of human hepatocytes represent assembled core particles and that monoclonal antibodies with known recognition sites can reveal region-specific core gene variation of the infecting HBV population. PMID- 9334924 TI - African strains of hepatitis E virus that are distinct from Asian strains. AB - Partial genomic sequences of four hepatitis E virus (HEV) strains from Africa (Morocco and Tunisia) and one from Central Asia (Tashkent, Uzbekistan) were obtained. The reverse transcriptase-polymerase chain reaction was used to amplify 5' and hypervariable regions of open reading frame 1 (ORF1) and a region overlapping all 3 ORFs. Sequence analysis of these regions revealed the African strains to be quite distinct from all known Asian strains but more similar to them than to the Mexican strain. Sequence analysis of the Tashkent strain revealed almost complete identity with another central Asian strain from Osh, Kirgizia. These results thus further confirm the geographical origin of HEV strain divergence. PMID- 9334925 TI - Contribution of tumor necrosis factor alpha and interleukin-1 alpha on the production of macrophage inflammatory protein-2 in response to respiratory syncytial virus infection in a murine macrophage cell line, RAW264.7. AB - The production of several inflammatory cytokines, such as murine macrophage inflammatory protein-2 (MIP-2), tumor necrosis factor (TNF), and interleukin (IL) 1, was investigated in response to respiratory syncytial virus (RSV) infection in a murine macrophage cell line, RAW264.7, with special reference to mutual relation of their productions. The kinetics of MIP-2 production showed a trend for a biphasic pattern, that is, MIP-2 levels became detectable from 2 h postinfection (p.i.) and increased markedly until 8 h p.i. Thereafter, this level fell to the same level until 16 h p.i. and then increased again. TNF alpha was also detectable at 2 h p.i. and then increased sharply until 8 h p.i., when the peak level attained. Compared with the levels of MIP-2 and TNF alpha, that of IL 1 alpha/beta, especially IL-1 beta, was lower (ng versus pg/ml order). The presence of anti-TNF alpha or anti-IL-1 alpha antibody did not influence the early phase of MIP-2 production but significantly inhibited the late phase, suggesting that MIP-2 is induced by the combined effects of RSV infection via direct induction and indirectly after initial induction of TNF alpha and IL-1 alpha productions. Although RSV-infected RAW264.7 cells had no alteration in viability compared with mock-infected control, these data demonstrate that RSV is a potent inducer of inflammatory cytokines by direct induction and indirectly via the initial production of other cytokines. PMID- 9334926 TI - The first human isolate of Puumala virus in Scandinavia as cultured from phytohemagglutinin stimulated leucocytes. AB - A virus isolate was recovered from blood leucocytes of a patient with nephropathia epidemica (NE). Leucocytes were isolated from EDTA-blood by dextran sedimentation and cultured on monolayers of Vero E6 cells in the presence of phytohemagglutinin (PHA) in roller tubes during the first 72 hours of incubation followed by rolling culture for three weeks in total. Thereafter the first subculture was done in a plastic flask and afterward at at least 6 week intervals. Antigen was first detected after 6 months and 2 weeks of culture. When tested by monoclonal antibodies and patient sera the isolate had the characteristics of a PUU virus. PCR amplification using PUU-specific primers and subsequent partial sequencing of the S and M segments revealed that the Umea/305/human/95 virus differs from the Finnish PUU Sotkamo rodent prototype virus and is similar but not identical to rodent strains of PUU virus acquired from the same region as the patient isolate. It is we concluded that the first human isolate of the etiologic agent of NE in Scandinavia was recovered from blood leucocytes stimulated with PHA by long-term culture in Vero E6 cells. The isolate belongs to the PUU serotype of hantaviruses as shown by its serologic profile and partial sequencing data. PMID- 9334927 TI - Infection with GB virus C and hepatitis C virus in drug addicts, patients on maintenance hemodialysis, or with chronic liver disease in Nepal. AB - Infection with GB virus C (GBV-C) and hepatitis C virus (HCV) was surveyed in various populations in Kathmandu, Nepal. GBV-C RNA and HCV RNA were detected in four (2%) and none, respectively, of 181 normal controls. Viral RNAs were detected significantly more frequently (P < 0.001) in 32 (44%) and 43 (60%), respectively, of 72 users of illicit intravenous drug, and in three (14%) and one (5%) of 22 patients on maintenance hemodialysis. The three hemodialysis patients with GBV-C RNA had been transfused with more blood units than the 19 without GBV C RNA (51 +/- 21 vs. 5 +/- 3 units, P < 0.01), and one was co-infected with HCV. Of 145 patients with chronic liver disease, GBV-C RNA was detected in four (3%) and HCV RNA in 12 (8%); only one patient with GBV-C RNA was without markers of HCV or hepatitis B virus infection. In the 32 drug addicts infected with GBV-C, genotypes were G1 in two (6%), G2 in 26 (81%), G3 in three (9%), and the remaining one (3%) was coinfected with G2 and G3. GBV-C genotypes in the 13 individuals in the populations other than drug addicts were G2 in 11 (85%) and G3 in two (15%). HCV genotypes in the 43 drug addicts with viremia were l/1a in 21 (49%), V/3a in 19 (44%) and l/1a plus V/3a in two (5%); these genotypes were not prevalent in normal controls and patients with chronic liver disease in Nepal. These results indicate that GBV-C infection is prevalent in healthy subjects in Nepal at a frequency (2%) comparable with those in the other countries and that GBV-C transmits efficiently by intravenous drug abuse among drug addicts and by transfusion in hemodialysis patients. PMID- 9334928 TI - High prevalence of hepatitis G viremia among kidney transplant patients in Thailand. AB - Patients receiving kidney transplants (KT) are at high risk for blood borne viral infections. To determine the prevalence of a recently discovered hepatitis G virus (HGV) in this patient group, reverse transcription-polymerase chain reaction (RT-PCR) employing primers derived from the NS5 region of the viral genome was utilized. HGV RNA was detected in 40 of 94 KT patients (43%), as compared to 3 of 69 healthy subjects (4.3%). Cocirculation of HGV and hepatitis C virus (HCV) RNA was detected in 12 patients (13%). Comparison of patients with and without HGV revealed that the former had received hemodialysis before transplantation for a significantly longer duration than the latter (28 vs. 17 months, respectively; P < 0.05). The amount of blood transfused and mean levels of liver enzymes, including alkaline phosphatase, alanine transaminase, and aspartate transaminase, were the same in both groups. Sequence analysis of 275 base pair DNA clones obtained from 2 patients revealed approximately 92% sequence homology to the published HGV and GB virus C sequences. These results suggested that HGV infection among Thai KT patients was high and the role of HGV in causing liver disease remains to be determined. PMID- 9334929 TI - Seroprevalence of GB virus C and persistence of RNA and antibody. AB - Exposure to GB virus C (GBV-C) was determined in several U.S. populations by both reverse-transcription-polymerase chain reaction (RT-PCR) and by an enzyme linked immunosorbent assay (ELISA) for antibodies to mammalian cell-expressed GBV-C envelope protein, E2 (GBV-C E2). Most individuals exposed to GBV-C were either RNA positive/ELISA negative or ELISA positive/RNA negative. Exposure, therefore, was measured as the sum of GBV-C RNA positive and GBV-C E2 antibody positive specimens, and was higher in commercial plasmapheresis donors (40.5%) than in volunteer blood donors (5.5%). In intravenous drug users (IVDUs), GBV-C exposure was 89.2%. Serial bleed specimens tested for GBV-C RNA indicate that some patients remain viremic for at least 3 years and fail to produce detectable antibodies to GBV-C E2. In other exposed individuals who tested negative for GBV C RNA, antibodies to E2 appear to be similarly long-lived (greater than 3 years) with a fairly constant titer (ranging in reciprocal endpoint dilution from 336 to 21,504). Since the detection of GBV-C RNA and GBV-C E2 antibody are mutually exclusive in most exposed individuals, studies pertaining to incidence and prevalence of GBV-C infection require both antibody and nucleic acid detection. PMID- 9334931 TI - Hepatitis C virus-induced leuko-thrombocytopenia and haemolysis. AB - Hepatitis C virus (HCV) has been recognized as the cause of thrombocytopenia occurring in patients with chronic hepatitis C, possibly through autoimmune mechanisms. A patient is described with B cell chronic lymphocytic leukaemia, presenting with a marked leuko-thrombocytopenia and an associated mild haemolysis secondary to HCV infection, in the absence of clinical and biochemical signs of hepatitis. Anti-HCV antibodies were detected in the serum both by ELISA and RIBA but not 2 months before the onset of cytopenia. The presence of HCV RNA was documented both in the peripheral blood mononuclear cells and in the bone marrow by reverse transcriptase polymerase chain reaction of the 5' untranslated region of the viral genome. Of interest, HCV RNA was also found in the serum, showing that viraemia was associated with the presence of circulating anti-HCV antibodies. HCV genotyping, performed by PCR amplification of the core region, revealed the presence of an unclassifiable genotype. The hypothetical mechanisms leading to HCV-induced cytopenia in this patient are briefly discussed. Treatment with corticosteroids was effective in controlling blood cell counts, without increasing viraemia and deterioration of liver disease. HCV infection should be considered in the differential diagnosis of possible causes of cytopenia, mainly in immunosuppressed patients, even in absence of clinical and biochemical signs of hepatitis. PMID- 9334930 TI - Puumala virus and two genetic variants of Tula virus are present in Austrian rodents. AB - Puumala and Tula viruses are hantaviruses found in Europe and are associated with the rodents Clethrionomys glareolus and Microtus arvalis, respectively. Puumala virus is associated with the human disease nephropathia epidemica. In Austria, ten clinically diagnosed cases of nephropathia epidemica, presumably caused by Puumala virus infection, have been reported but not virologically confirmed [Leschinskaya et al., 1991; Aberle et al., 1996]. To identify the hantaviruses that are present in Austria, five species of rodents were trapped and screened for virus antibodies, antigen, and RNA. Hantaviruses were detected in two species, Cl. glareolus and M. arvalis, by reverse transcription-polymerase chain reaction (RT-PCR). RT-PCR products from Cl. glareolus tissues yielded a unique Puumala virus sequence distinct from Puumala virus sequences reported from other parts of Europe. RT-PCR products from M. arvalis tissues yielded two genetically distinct Tula virus sequences, one similar to sequences reported from Slovakia and the Czech Republic and another that appears to be a novel genetic variant of Tula virus. This is the first confirmed report of hantaviruses in Austria. PMID- 9334932 TI - The rate of aldehyde fixation of the exocytotic machinery in cultured hippocampal synapses. AB - The rate at which the transmitter release machinery is fixed by 2% glutaraldehyde at hippocampal synapses and the amount of release evoked by the fixative have been investigated. We recorded from hippocampal cells while fixative was applied with a rapid flow system. Release is blocked in less than a second and fixative produced exocytosis is at most a few percent of what would be caused by a hypertonic stimulus that completely depletes the readily releasable pool of vesicles. The postsynaptic receptors for glutamate cease to respond to agonist with a time constant of approximately 3 s when fixative is applied. We conclude that some essential component of the exocytotic apparatus is fixed in less than a second and that the fixative does not significantly deplete the readily releasable pool. PMID- 9334933 TI - Dual-channel telemetry system for recording vocalization-correlated neuronal activity in freely moving squirrel monkeys. AB - A miniature telemetric system is described which allows simultaneous measurements of neural activity and vocalization in freely moving monkeys within their social group. Single and multi-unit activities were detected with medium impedance electrodes that were fixed to self-made microdrives allowing accurate vertical positioning over a range of 8 mm. Vocalizations were registered by means of a piezo-ceramic device sensing the vocalization-induced skull vibrations. This allowed identification of the vocalizing animal in a larger group and eliminated environmental noise. Neuronal activity and vocalization were transmitted via separate channels of a FM transmitter using different carrier frequencies. The signals were decoded in two conventional FM receivers equipped with an automatic frequency control. The signals were stored for off-line analysis on a HiFi videotape recorder. PMID- 9334934 TI - Zymographic measurement of gelatinase activity in brain tissue after detergent extraction and affinity-support purification. AB - Several methods have been developed for the measurement of gelatinase activity from various tissues using detergent extraction. Gelatin-affinity chromatography has been employed for the large-scale purification of gelatinases from conditioned medium obtained from cultured cells. The objective of this paper was to develop a rapid method whereby gelatinase activity could be extracted from regional brain tissues without tedious, intervening purification steps. After Triton X-100 extraction and gelatin-Sepharose 4B purification of rat brain tissue extracts, two major activities were observed on gelatin zymograms. These were identified as gelatinase A and B using co-migration with astrocyte-derived enzymes and inhibition of activity by tissue inhibitor of matrix metalloproteinase-1 (TIMP-1). The non-ionic detergents, Triton X-100 and 3-[(3 cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS) were equally effective in extracting activities from brain tissue. Little difference in recovery was observed among 0.1, 1 and 10% concentrations of Triton X-100. The method developed here was capable of recovering gelatinase activities from rat brain tissue over a 4-10-fold range using gelatin zymography for the measurement of activity. It is possible that this method may be modified for the measurement of gelatinases in tissues such as biopsy samples of gliomas or astrocytomas or other cancers where gelatinases are thought to play a role in tumor invasion and/or metastasis. PMID- 9334935 TI - Use of differential normal pulse voltammetry for the measurement of locus coeruleus catecholaminergic metabolism in an acute anaesthetized rodent model of allodynia: effect of mexiletine. AB - Neuropathic pain can be triggered by non-painful stimuli (e.g., light touch), a sensory abnormality termed allodynia. The acute blockade of spinal glycine receptors with intrathecal strychnine induces a reversible allodynia-like state in the rat. We describe the application of in vivo differential normal pulse voltammetry with carbon fibre micro-electrodes for monitoring the catechol oxidation current (CAOC) of the locus coeruleus (LC) in the strychnine model of allodynia. In addition, we tested the effect of mexiletine, a drug useful in the management of clinical neuropathic pain in this model. Our results show that somatosensory processing in the spinal cord of urethane-anaesthetized rats is radically altered during glycine receptor blockade such that the normally innocuous stimulus of hair deflection causes the marked activation of the LC as determined using in vivo differential normal pulse voltammetry. Mexiletine suppressed the LC and cardiovascular responses of strychnine induced allodynia. Results of this study indicate that LC CAOC, an index of LC neuronal activity: (a) is a sensitive biochemical index of strychnine-allodynia; (b) is temporally correlated with the cardiovascular and motor responses evoked by hair deflection during glycine receptor blockade; and (c) can be used to quantitate allodynia in the strychnine model. PMID- 9334936 TI - A simple water-immersion condenser for imaging living brain slices on an inverted microscope. AB - Due to some physical limitations of conventional condensers, inverted compound microscopes are not optimally suited for imaging living brain slices with transmitted light. Herein is described a simple device that converts an inverted microscope into an effective tool for this application by utilizing an objective as a condenser. The device is mounted on a microscope in place of the condenser, is threaded to accept a water immersion objective, and has a slot for a differential interference contrast (DIC) slider. When combined with infrared video techniques, this device allows an inverted microscope to effectively image living cells within thick brain slices in an open perfusion chamber. PMID- 9334937 TI - Quantitative measurement of cerebral protein synthesis in vivo: theory and methodological considerations. AB - The true rate of cerebral protein synthesis can be calculated from the ratio of labeled proteins to integrated arterial plasma amino acid specific activity (SA) only when the fraction of amino acid precursor pool dilution is known. In the following, current experimental designs on the measurement of cerebral protein synthesis are discussed and compared to our own approach in which the determination of regional precursor pool dilution by recycled unlabeled leucine is combined with the quantitation of regional cerebral protein synthesis rates. For this purpose, a constant arterial plasma leucine SA level is maintained for 45 min by programmed intravenous infusion which is sufficient for complete equilibrium between tissue leucine pool SAs and plasma free leucine SA. In addition to the regional assessment of the precursor dilution factor, protein radioactivity can be determined in the same tissue sample or in parallel brain sections of the same animal by quantitative autoradiography. It is then possible to calculate the actual rate of protein synthesis using the correct fraction of precursor pool dilution. This renders our approach particularly suitable for the quantitative measurement of regional CPS under pathological conditions. PMID- 9334939 TI - Increased amount of zinc in the hippocampus and amygdala of Alzheimer's diseased brains: a proton-induced X-ray emission spectroscopic analysis of cryostat sections from autopsy material. AB - Zinc has been implicated as a contributing cause of the neuropathology of Alzheimer's disease (AD), but consensus on the zinc content of AD brains has not yet been established. In the present study, multi-element PIXE was used to measure zinc in cryostat sections of brain tissue from AD patients and from normal control subjects. Compared to their age-matched controls, the AD patients showed an increase in zinc in the hippocampal and amygdalar regions. The instrumental PIXE assays do not show whether the zinc changes are due to altered zinc in the boutons of Zinc-ENriched (ZEN) neurons, i.e., zinc ions in synaptic vesicles, or to changes in the amount of zinc tightly bound to macromolecules. We hypothesise that the increased zinc level is caused by an increase in the amount of ZEN terminals. Such an increase could be the result of a sprout of ZEN terminals in diseased areas of the brain. PMID- 9334938 TI - In vivo saturation kinetics of two dopamine transporter probes measured using a small animal positron emission tomography scanner. AB - When estimated in vitro, the parameters which describe the binding of radiolabelled analogues of cocaine to sites on the dopamine transporter are very much influenced by the methodology used. In the present study, a small animal positron emission tomography (PET) scanner was used to estimate in vivo saturation kinetics for two carbon-11 labelled compounds presently used to monitor dopamine terminal function. The binding of [11C]CFT (WIN 35,428) in rat striatum was adequately described by a single-site model, giving an apparent dissociation constant corresponding to an intravenous dose of 242 nmol/kg. In contrast, the binding of [11C]RTI-121 was better described by a two-site model with the 'high-affinity' site or state (dissociation constant = 1 nmol/kg) being significantly occupied at doses routinely used in PET scanning. Such findings cannot readily be predicted from in vitro work, but could aid in both the choice of ligand and the model used in quantification of scan data. While multi-dose in vivo PET studies are difficult in man, rat PET can easily be employed either pre clinically for putative radioligands, or experimentally, to study drug interactions and receptor occupancy related to functional efficacy. PMID- 9334940 TI - A semi-in-vivo preparation for optical recording of the insect brain. AB - We describe a method for optically recording neuronal activity from an intact insect brain, upon natural sensory stimulation. In this preparation, the head capsule of the honeybee, Apis mellifera, is isolated from the body while leaving the entire brain undamaged. In short, a hole is first cut into the cuticule to allow optical access to the brain and to allow the removal of tracheae and glands. Then the head is cut free and placed into a dye-loaded and cooled ringer solution in a staining chamber for 1 h. Subsequently, the head is fixed in a recording chamber, covered with a cover-slip, and imaged under the microscope with a cooled CCD camera. The whole preparation leaves the antennae dry, free to move, and functional throughout the experiment, allowing for natural odour stimulation of the olfactory system. Using calcium sensitive or potential sensitive dyes (calcium-green or RH795), we could record the processing of olfactory information at the glomerular level in the antennal lobe of the bee. PMID- 9334941 TI - An electromyographic technique for small animals. AB - An improved technique is proposed to record electromyographic (EMG) signals. The technique includes anesthesia with isofluothane and twisted bipolar electrodes that are glued with a sugar solution to longitudinally grinded hypodermic needles. The advantages of the technique are (1) minimal damage to the muscles; (2) removal of the injection needle with ease after insertion into the muscle; (3) the electrodes can be soldered onto the connector prior to the experiment; and (4) quick recovery of the animal. The technique is especially advantageous for EMG experiments with small animals. PMID- 9334942 TI - Commercial mouse and human nerve growth factors contain nerve growth factor prohormone isoforms. AB - Highly sensitive chemiluminescence immunoblot analysis was utilized to examine the purity of mouse 2.5S-, beta- and 7S nerve growth factors as well as that of recombinant human beta-nerve growth factor obtained from commercial vendors. Three polyclonal antisera and two monoclonal antibodies to 13 kDa nerve growth factor (2.5S NGF and beta-NGF) were employed for assessing the purity of each preparation. In addition, polyclonal antisera against two prepro-NGF specific domains were used for immunoblotting analysis to ascertain the identity of high molecular weight nerve growth factor immunoreactive proteins as prohormones. Both the mouse and human NGF preparations contained 53 and 60 kDa immunoreactive proteins. Of these, the mouse 60 kDa and the human 53 kDa proteins strongly immunoreacted with both prepro-nerve growth factor specific domain antibodies suggesting that they are two NGF prohormone isoforms. In addition, both the mouse and human nerve growth factor preparations contained proteins that were immunoreactive to polyclonal antisera and monoclonal antibodies to mouse 2.5S and/or beta-NGF. High molecular weight aggregates of prohormones were also observed in mouse and human nerve growth factor samples. In summary, none of the ten NGF samples examined were pure as stated. Our study cautions investigators in the field to be aware of the presence of nerve growth factor prohormones and other proteins in various mouse and human nerve growth factors sold commercially. PMID- 9334943 TI - Applications of multimedia computers and video mixing to neuroethology. AB - Inexpensive multimedia computers offer new possibilities for mixing video and computer images, videotaping these mixed images, and extracting quantitative data from videotape. In this paper we describe methods for mixing images from a video camera and a Macintosh computer display using chroma keying, and we describe a simple circuit for analog video mixing and frame-counting. We present three applications of these video mixing methods to our neurophysiological and behavioral research with awake, behaving animals. These technologies enhance accuracy, speed, and flexibility during experiments, by allowing us to record an information-rich videotape of the subject and state of the experimental apparatus. After the experiment, these technologies facilitate selecting and extracting quantitative data from the videotape for further analysis. The videotape is especially useful in resolving minor inconsistencies or incomplete information in the notes and data files that often arise during analysis of complex experiments. PMID- 9334944 TI - Excitability of neural elements within the rat corpus striatum. AB - The excitability of cholinergic, glutamatergic and dopaminergic elements within the rat neostriatum was studied in both in vivo and in vitro preparations. In vivo, the microdialysis technique was used to measure the release of striatal acetylcholine and dopamine under basal and electrically evoked conditions. For comparison, acetylcholine, dopamine and glutamate release was assayed in media obtained from superfused rat striatal slices. Electrical stimulation was used to derive the strength-duration functions and their chronaxies of stimulated elements containing the three neurotransmitter types. The chonaxies for experiments in vitro and in vivo were similar: the chronaxy values for elements containing acetylcholine were the shortest, the values for glutamate were intermediate, and the values for those containing dopamine were the longest. Based on the chronaxy estimates, it is proposed that the elements containing acetylcholine are the large cholinergic interneurons of striatum, and the elements containing glutamate and dopamine are the terminals of corticostriatal and nigrostriatal neurons, respectively. These results indicate that electrical stimulation of neural elements surrounding a microdialysis probe can be an additional tool to examine the factors that regulate neurotransmitter release. Likewise, investigators can activate specific striatal elements by using pulse durations that coincide with their chronaxies. PMID- 9334945 TI - Implants for sustained drug release over the somatosensory cortex of the newborn rat: a comparison of materials and surgical procedures. AB - Drugs that interfere with neural transmission are an important tool in assessing the role of specific neurotransmitters in the development of the nervous system. Systemic drug treatments often produce neurodevelopmental effects with questionable specificity. Furthermore, many compounds of interest do not cross the blood-brain barrier. To overcome these limitations, either elvax or gelfoam implants have been previously employed to produce sustained drug release over specific brain regions. In this paper, stereotaxic coordinates are provided for reproducible insertion of drug-delivery systems over the rat somatosensory cortex at birth (P0), prior to the appearance of the cortical barrel pattern; a novel and simpler method for preparation of elvax 40p sheets is described; a new implantation technique is provided. Furthermore, we compare the efficiency and tolerability of elvax vs gelfoam implants, showing that gelfoam, but not elvax, significantly disrupts cortical cytoarchitecture. Finally, successful destruction of serotonin-containing terminals in layer IV of the primary somatosensory cortex of the newborn rat is demonstrated by application of parachloroamphetamine containing elvax implants. PMID- 9334946 TI - Interobserver variation in the AO/OTA fracture classification system for pilon fractures: is there a problem? AB - OBJECTIVES: To evaluate the interobserver variation for the AO/OTA fracture classification system: region forty-three-pilon fractures. METHODS: One senior attending, two fellows (one trauma, one foot and ankle), one junior orthopaedic resident, and one experienced research coordinator independently classified eighty-four sets of radiographs. The evaluator was blinded as to treatment and functional outcome. The radiographs initially used to manage the patients were evaluated; no special radiographs or standardized radiographic techniques were used. The kappa statistic, Williams index, and SAV statistic were calculated. RESULTS: Using the SAV statistic to quantify rater agreement beyond that expected by chance alone, the average chance-adjusted agreement among the raters was 0.57 for fracture type, 0.43 for group, and 0.41 for subgroup. This is equivalent to moderate agreement (0.41 to 0.60). The kappa statistic was used to determine whether there was difficulty with any specific category of the AO type classification among raters for selecting fracture type (A, B, C). Kappa values were 0.49 for type A, 0.58 for type B, 0.57 for type C, all of which were considered adequate. CONCLUSION: These data are similar to others reported for interobserver agreement with the AO/OTA fracture classification and other classification systems. The issue of individual judgement in taking a continuous variable (fracture pattern) and compartmentalizing it into a dichotomous variable (fracture classification system) is highlighted by these data. Determination of fracture types alone (type A, B, or C) would seem to be sufficient for clinical research where fracture severity should be reported as a variable. PMID- 9334947 TI - A critical assessment of factors influencing reliability in the classification of fractures, using fractures of the tibial plafond as a model. AB - OBJECTIVE: To investigate three factors that may influence the reliability of a fracture classification system: (a) the quality of the radiographs; (b) the ability of observers to identify the fracture fragments; and (c) the use of binary decision making. DESIGN: Assessment of interobserver reliability of blinded observers. SETTING: Medical school department of orthopaedics. PARTICIPANTS: Two attending orthopaedists, two PGY-5 orthopaedic residents, and two PGY-3 orthopaedic residents served as observers. INTERVENTION: Observers classified radiographs of twenty-five tibial plafond fractures according to the Ruedi-Allgower and binary classification systems, and also rated the quality of each radiograph as adequate or inadequate for accurately classifying the fracture. At a second session, observers classified the same radiographs after marking the fragments of the tibial articular surface, as well as radiographs that had the articular fragments premarked by the senior author. MAIN OUTCOME MEASURES: Pairwise interobserver reliability was analyzed by kappa statistics, and mean kappa values were compared for each method of fracture classification. RESULTS: No difference in interobserver reliability was detected between the Ruedi-Allgower and binary classification systems. Interobserver agreement on the adequacy of the radiographs was poorer than agreement on the classification of the fractures themselves. Having observers mark the fragments of the tibial articular surface had no effect on interobserver reliability; having the articular fragments premarked, however, significantly improved interobserver reliability in classifying the fractures. CONCLUSIONS: The results of this study underscore the complexity of tibial plafond fractures and the difficulty observers have in reliably interpreting fracture radiographs. Fracture classification systems, such as the Ruedi-Allgower, predicated on identification of the number and displacement of articular fragments, may inherently perform poorly on reliability analyses because of observer difficulty in reliably identifying the fragments. Because binary decision making did not improve the reliability of fracture classification in this study, further investigation of the effectiveness of binary decision making may be advisable before such strategies are put into widespread use. PMID- 9334948 TI - Assessment of the AO/ASIF fracture classification for the distal tibia. AB - OBJECTIVES: The purpose of this study was to assess the interobserver reliability and intraobserver reproducibility of the AO/ASIF and Ruedi and Allgower classifications for fractures of the distal tibia, and to determine the benefit of a computed tomography (CT) scan and experience on observer agreement for several fracture characteristics, including classification. METHODS: The radiographs of forty-three fractures of the distal tibia, fourteen of which had CT scans, were assessed by groups of experienced and less-experienced observers. Each case was classified according to the AO/ASIF and Ruedi and Allgower systems. Several other fracture characteristics also were assessed. The kappa coefficient of agreement was calculated and used to compare the interobserver reliability and intraobserver reproducibility of the classification systems and to determine the benefit of experience and CT scans. The intraclass correlation coefficient was used to assess noncategoric data. RESULTS: Interobserver and intraobserver agreements were good when classifying fractures into AO/ASIF types and significantly better than that for the Ruedi and Allgower system. However, agreement was poor when classifying the fractures into AO/ASIF groups. For most assessments, the experienced group tended to have higher levels of interobserver agreement, but not intraobserver agreement. Viewing the CT scans improved agreement on the percentage of articular surface involved, but it did not improve interobserver reliability or intraobserver reproducibility for either of the classification systems. CONCLUSION: The AO/ASIF classification for fractures of the distal tibia has good observer agreement at the type level, but poor agreement at the group level. Experience tends to improve interobserver agreement, but not intraobserver agreement. Viewing CT scans does not improve agreement on classification, but it tends to improve agreement on articular surface involvement. PMID- 9334949 TI - Impact of CT scan on treatment plan and fracture classification of tibial plateau fractures. AB - OBJECTIVE: To evaluate the interobserver and intraobserver agreement for both treatment plan and fracture classification of tibial plateau fractures using plain films alone and with computed tomography (CT) scans. DESIGN: Prospective study to assess the impact of an advanced radiologic study on the agreement of treatment plan and fracture classification of tibial plateau fractures. SETTING/PARTICIPANTS: Two orthopaedic traumatologists, two orthopaedic residents, and two skeletal radiologists were presented with twenty-one cases of tibial plateau fractures imaged with plain films and with CT scans. MAIN OUTCOME MEASURES: Agreement was measured using kappa coefficients. RESULTS: Using plain films alone, the mean interobserver kappa coefficient for classification was 0.62, which decreased to 0.61 after addition of CT scans. Using plain films alone for formulating a treatment plan, the mean interobserver kappa coefficient was 0.58, which increased to 0.71 after addition of CT scans. The mean intraobserver kappa coefficient for fracture classification using plain films was 0.70, which increased to 0.80 with addition of CT scans. The mean intraobserver kappa coefficient for treatment plan based on plain films alone was 0.62, which increased to 0.82 after addition of CT scans. Class was changed in an average of 12 percent of cases after addition of CT scans. Treatment plan was changed an average of 26 percent of the time after addition of CT scans. CONCLUSION: Addition of CT scans to plain roentgenograms increases the interobserver and intraobserver agreement on treatment plan. PMID- 9334950 TI - Effect of stainless steel and titanium low-contact dynamic compression plate application on the vascularity and mechanical properties of cortical bone after fracture. AB - OBJECTIVES: Comparison of the effect of stainless steel and titanium low-contact dynamic compression plate application on the vascularity and mechanical properties of cortical bone after fracture. DESIGN: Randomized, prospective. SETTING: Orthopaedic research laboratory. ANIMALS: Ten large (greater than twenty five kilogram) adult dogs. INTERVENTION: A short, midshaft spiral tibial fracture was created, followed by lag screw fixation and neutralization with an eight hole, 3.5-millimeter, low-contact dynamic compression plate (LCDCP) made of either 316L stainless steel (n = five) or commercially pure titanium (n = five). After surgery, animals were kept with unrestricted weight-bearing in individual stalls for ten weeks. MAIN OUTCOME MEASUREMENTS: Cortical bone blood flow was assessed by laser Doppler flowmetry using a standard metalshafted probe (Periflux Pf303, Perimed, Jarfalla, Sweden) applied through holes in the custom-made LCDCPs at five sites. Bone blood flow was determined at four times: (a) prefracture, (b) postfracture, (c) postplating, and (d) ten weeks postplating. After the dogs were killed, the implant was removed and both the treated tibia and contralateral tibia were tested for bending stiffness and load to failure. RESULTS: Fracture creation decreased cortical perfusion in both groups at the fracture site (p = 0.02). The application of neither stainless steel nor titanium LCDCPs further decreased cortical bone blood flow after fracture creation. However, at ten weeks postplating, cortical perfusion significantly increased compared with acute postplating levels in the stainless steel (p = 0.003) and titanium (p = 0.001) groups. Cortical bone blood flow ten weeks postplating was not significantly different between the titanium group and the stainless steel group. Biomechanical tests performed on the tibiae with the plates removed did not reveal any differences in bending stiffness nor load required to cause failure between the two groups. CONCLUSIONS: Both titanium and stainless steel LCDCPs were equally effective in allowing revascularization, and neither provided a significant advantage in biomechanical properties of fracture healing at ten weeks. PMID- 9334951 TI - Bovine-derived bone protein as a bone graft substitute in a canine segmental defect model. AB - OBJECTIVES: To evaluate the efficacy of a bone graft substitute in healing of a segmental defect of a weight-bearing long bone. DESIGN: An established canine model was used to perform a blinded, prospective, randomized study of the performance of bone graft substitute implants. This performance was compared with that of an accepted treatment modality (autograft) in a paired fashion. SETTING: An accredited animal research facility. SUBJECTS AND INTERVENTION: Twenty-eight dogs underwent bilateral radial osteotomies with creation of a 2.5-centimeter defect. On one side, the defect in every dog was filled with autogenous cancellous bone graft (ABG). Contralateral defects received, in a blinded, randomized fashion, cylindrical implants of demineralized bone matrix (DBM) allograft or DBM plus a constant dose (3.0 milligrams) of bovine-derived bone protein (DBM + BP). The defects were stabilized by external fixation. Subjects underwent monthly radiographs and were killed at six, twelve, or twenty-four weeks. Regenerate bone was studied by biomechanical testing and histology. Six animals were studied to determine the dose-response characteristics of the protein preparation. Three received implants containing 0.3 milligram of BP (group 1) and three received 1.0 milligram of BP (group 2). These animals were killed at twelve weeks of follow-up. RESULTS: All twenty-eight ABG radii (100 percent) progressed to radiographic union, as did thirteen of thirteen (100 percent) DBM + BP radii compared with only four of fifteen (27 percent) of DBM radii. The difference between union rates was statistically significant (p < 0.05). Mean values for most biomechanical parameters of DBM + BP radii exceeded those of their contralateral ABG controls at twelve and twenty-four weeks, whereas those for DBM implants did not. Histology revealed microscopic evidence of normal bone healing in all ABG and DBM + BP radii, whereas most DBM radii demonstrated nonunions. In the dose-response arm of the study, six of six ABG radii (100 percent) achieved union; zero of three (0 percent) of group 1 and two of three (67 percent) of group 2 radii achieved grossly stable unions. Biomechanical testing was consistent with radiographic results, indicating that the 3.0-milligram dose was the most effective of those studied. CONCLUSIONS: The DBM + BP composite implants were more effective at healing critical-sized segmental defects than DBM alone in this canine model when a 3.0-milligram per implant dose of BP was used. Biomechanical and histologic properties of the regenerated bone formed by DBM + BP implants was comparable to that of cancellous autograft. PMID- 9334952 TI - Associated lumbosacral junction injuries (LSJIs) in pelvic fractures. AB - OBJECTIVE: To study lumbosacral junction injuries (LSJIs) associated with displaced sacral fractures. DESIGN: Prospective. SETTING: University hospital. PATIENTS: Eighty-nine patients with pelvic fractures. INTERVENTIONS: All patients underwent standard x-ray examination consisting of anteroposterior and inlet and outlet views of the injury. In 50 percent of the cases, oblique views were also obtained. Computed tomography (CT) scans were obtained for all vertically unstable lesions and for fifteen of the lateral compression lesions. MAIN OUTCOME MEASURES: Pelvic fractures were classified according to Tile's classification. LSJIs associated with pelvic ring fractures were classified according to Isler. RESULTS: Lumbosacral junction injuries, including one bilateral lesion, were found in thirteen patients. One lesion was associated with a compression fracture, and the other twelve were associated with vertically unstable fractures. In one patient, a facet fracture was associated with a contralateral sacroiliac joint dislocation. A new type of LSJI was identified in three patients: it was characterized by disruption of the annulus fibrosus associated with rising of the hemipelvis and inclination of the L5 body. CONCLUSION: Lumbosacral junction injuries should be suspected in cases of transforaminal sacral fracture, especially when these fractures are displaced. In such cases, we recommend that the lumbosacral junction be evaluated with appropriate CT scans. PMID- 9334953 TI - The management of femoral diaphyseal nonunions. AB - OBJECTIVE: To assess the efficacy of treatment and develop an algorithm for management of nonunions of the femoral diaphysis. STUDY DESIGN: Retrospective. SETTING: University hospital. METHODS: Forty-four patients treated at one institution for nonunion of the femoral diaphysis were studied. Thirteen of these patients had a history of infection. After debridement (where appropriate) and repair of the femoral nonunion, follow-up averaged twenty-eight months (range, 24 to 108 months). All patients were examined at final follow-up. RESULTS: Thirty three patients achieved union after one procedure, and eight patients achieved union after additional procedures. One patient underwent above-knee amputation, and two patients remained ununited at the time of their final follow-up. Time to union averaged 11.8 months. Seventeen patients healed with more than two centimeters of shortening, and ten patients lost more than 30 degrees of knee flexion. CONCLUSION: Established femoral diaphyseal nonunions can be treated effectively, even in the presence of chronic sepsis. Selective use of a vascularized fibula transfer has proven beneficial in addressing intercalary defects. Plate fixation, with or without a vascularized fibula transfer, has been the predominant mode of skeletal stabilization in more complex reconstructions. PMID- 9334954 TI - Distal femoral fixation: a biomechanical comparison of the standard condylar buttress plate, a locked buttress plate, and the 95-degree blade plate. AB - OBJECTIVES: This biomechanical cadaver study was performed to compare the fixation stability of a standard lateral condylar buttress plate with a similar condylar buttress plate with the distal screws locked to the plate. Then the study was repeated with six additional matched femoral pairs to compare the locked plate with a standard 95-degree blade plate. DESIGN: Six matched pairs of mildly osteopenic femurs were selected, and each side was assigned randomly to fixation with either a standard lateral condylar buttress plate or a modified lateral condylar buttress plate with locked distal screws. The experiment was repeated with six additional matched pairs of femurs instrumented with either a modified lateral condylar buttress plate with locked distal screws or a standard 95-degree blade plate. INTERVENTION: The femurs were instrumented, and a gap osteotomy was created at the distal femoral metaphysis. The instrumented femurs were then mechanically tested in axial compression and bending/torsional loading to determine fixation stability; then they were loaded at 1,000 newtons for 10(5) cycles and retested for stability. MAIN OUTCOME MEASUREMENT: The displacement across the osteotomy gap at 100-newton and 1,000-newton axial loads was measured directly for each specimen before and after cycling. In addition, resistance to displacement in bending/torsional loading (newtons/centimeter) was determined from load/displacement curves, before and after cycling. RESULTS: The locked buttress plate provided significantly greater fixation stability than the standard plate both before and after cycling in axial loading. The locked buttress plate also proved significantly more stable in axial loading than the blade plate both before and after cycling. CONCLUSION: A condylar buttress plate with locked screws is a valid concept for improving fixation stability. PMID- 9334955 TI - Knee dislocation: initial assessment and implications for treatment. AB - OBJECTIVE: To evaluate bicruciate knee injuries and determine whether they should be treated as knee dislocations, especially with regard to vascular injuries. DESIGN: Retrospective. SETTING: University hospital, level 1 trauma center. PATIENTS: Fifty patients admitted between 1987 and 1994 who had sustained knee dislocations or bicruciate ligament injuries. MAIN OUTCOME MEASURES: Mechanism of injury, direction of dislocation, knee ligament injury pattern, presence or absence of periarticular fracture, presence of vascular and nerve injuries, and location of associated trauma were measured. RESULTS: Twenty-two knees had classic knee dislocations. Twenty-eight knees presented as "reduced" bicruciate ligament injuries. Vascular injury occurred just as frequently in bicruciate ligament injuries as in knee dislocations. The direction of the knee dislocation did not predict ligament injury pattern or the presence of arterial injury. CONCLUSION: Bicruciate ligament injuries are equivalent to knee dislocations with regard to mechanism of injury, severity of ligamentous injury, and frequency of major arterial injuries. PMID- 9334956 TI - Acute repair and delayed reconstruction for lateral ankle instability: twenty year follow-up study. AB - OBJECTIVES: To determine long-term results of patients who underwent primary ligament repair and delayed reconstruction for lateral ligament instability. DESIGN: Retrospective. SETTING: Outpatient clinic. PATIENTS/PARTICIPANTS: Patients who had undergone acute repair or delayed reconstruction at this institution between 1958 and 1977, excluding patients who were deceased or who could not be located. INTERVENTION: Forty-eight patients (fifty-three ankles) underwent twenty-two primary ligament repairs and thirty-one delayed reconstruction operations. MAIN OUTCOME MEASUREMENTS: Clinical results graded with clinical scale and radiologic results based on stress radiographs and plain film radiographs. RESULTS: At an average of twenty years after operation (range 12 to 33 years), patients were satisfied with forty-nine ankles, satisfied with reservations with two ankles, and dissatisfied with two ankles. Clinical results after repair were excellent in twenty ankles, good in one, fair in none, and poor in one. After reconstruction, the results were excellent in twenty-one ankles, good in six, fair in one, and poor in three. In the primary repair group, the mean talar tilt with stress testing improved from 20.7 +/- 10.7 degrees before operation to 2.8 +/- 3.0 degrees after operation. In the reconstruction group, the mean talar tilt improved from 20.7 +/- 8.4 degrees before operation to 2.8 +/ 3.5 degrees after operation. CONCLUSIONS: Clinical and radiologic results were similar in the repair and reconstruction groups. The majority of severe (Grade III) ankle sprains may be treated nonoperatively, but if residual instability occurs, late reconstruction should achieve satisfactory results. PMID- 9334957 TI - External fixation for severe open fractures of the humerus caused by missiles. AB - OBJECTIVE: To evaluate the use of external fixation of the humerus after missile injuries. DESIGN: Retrospective. SETTING: University medical center. PATIENTS: Twenty-six soldiers with twenty-six open Gustilo type III fractures. INTERVENTIONS: Immediate external fixation. MAIN OUTCOME MEASURES: Clinical, functional, social, and rehabilitation criteria were evaluated. RESULTS: Excellent in fourteen patients (61%), good in four (17%), fair in three (13%), and poor in two (9%). All fractures eventually healed. CONCLUSION: External fixation is the preferred initial treatment for stabilizing severe open missile fractures of the humerus. Its use, together with radical debridement of dead bone, has reduced the incidence of chronic infection and improved the prognosis of vascular repairs. As a result, the rate of morbidity and upper limb amputation has been reduced significantly, compared with our previous experience. PMID- 9334958 TI - Intrapelvic dislocation of the left hemipelvis as a complication of the pelvic "C" clamp: a case report and review. AB - High-energy pelvic trauma, with posterior pelvic disruption, produces high morbidity and mortality rates. Part of the initial resuscitation has included an anterior external fixator to close the pelvic ring, thereby decreasing blood loss and reducing mortality. However, this technique has been found to be less efficacious in certain situations. This has stimulated an interest in alternative methods of stabilization, which has led to the recent development of the emergency pelvic "C" clamp. We present one of the potential pitfalls of this new device, discuss pertinent clinical and biomechanical studies, and offer suggestions regarding its use. PMID- 9334986 TI - Effect of CO on VO2 of carotid body and chemoreception with and without Ca2+. AB - This study was done using high PCO (> 500 Torr at PO2 of 120 Torr) in the carotid body perfusate in vitro, and recording simultaneously the activity of the whole carotid sinus nerve (CSN) and VO2 of the carotid body. In the cascade of excitation of CSN by high PCO in the dark [light eliminated the excitation; S. Lahiri, News Physiol. Sci. 9 (1992) 161-165], Ca2+ effects occur at the level of neurosecretion after the level of oxygen consumption, according to the following scheme: CO-hypoxia-->VO2 decrease-->K+ conductance decrease-->cell depolarization ->cytosolic Ca2+ rise-->neurosecretion-->neural discharge. Thus, a part of the hypothesis was that [Ca2+] decrease, being a downstream event, may not affect VO2 of the carotid body. Also, to determine to what extent the intracellular calcium stores contribute to cystolic [Ca2+] and chemosensory discharge with high PCO, we tested the effect of interruption of perfusate flow with medium nominally free of [Ca2+] on CSN excitation and VO2 of the carotid body with and without high PCO. High PCO in the dark decreased carotid body VO2, independent of [Ca2+]o. CSN excitation was always enhanced by high PCO, and its sensitivity to perfusate flow interruption. Also, nominally Ca(2+)-free solution increased the latency and decreased the rate of rise and peak activity of CSN during interruption of perfusate flow, but CO augmented the responses. This reversal effect by CO suggests that Ca2+ is released from intracellular stores, because CO has no other way to excite the chemoreceptors than by acting on the intracellular stores. PMID- 9334987 TI - Changes in regional blood flow in response to distension of the uterus in anaesthetised pigs. AB - The present study was undertaken in anaesthetised pigs to determine the primary reflex effects of distension of the uterus on the peripheral circulation. Experiments were performed in seven pigs anaesthetised with alpha-chloralose and artificially ventilated. Blood flow in the superior mesenteric, left renal and left external iliac arteries was assessed using electromagnetic flowmeters. Distension of the uterus was performed whilst preventing changes in heart rate and aortic blood pressure by injecting 20 ml of warm Ringer solution in a balloon positioned within the viscus (mean transmural pressure of about 18 mmHg). In each pig, distension of the uterus caused decreases in all measured blood flows. In four pigs, these decreases were graded by step increments of distension. In the seven pigs, the responses of decrease in mesenteric, renal and iliac blood flows were not affected by blockade of beta-adrenergic receptors with propranolol, but were abolished by the subsequent blockade of alpha-adrenergic receptors with phentolamine. The present study showed that distension of the uterus in anaesthetised pigs primarily caused reflex vasoconstriction in the mesenteric, renal and iliac vascular beds. This reflex response was mediated by sympathetic mechanisms which involved alpha vascular adrenergic receptors. PMID- 9334988 TI - Oxytocin decreases blood pressure in male but not in female spontaneously hypertensive rats. AB - The aim of the present study was to investigate the effects of repeated injections of oxytocin on blood pressure and heart rate in spontaneously hypertensive rats (SHR). For this purpose subcutaneous (s.c.) injections of oxytocin 1 mg/kg or saline were given for 5 days to male and female SHR. Blood pressure and heart rate were measured daily before, during and after the oxytocin treatment period. In male rats, a significant decrease in blood pressure (systolic; p < 0.01, diastolic; p < 0.05), but no effect on heart rate, was seen the day after the first injection of oxytocin, when compared to saline-treated controls. Blood pressure decreased further in response to each injection and a maximal difference of 21 mmHg (systolic) (p < 0.01), compared to controls, was reached after the last injection. The significant effect was gone 3 days after the last injection, although a tendency to a lower blood pressure in the oxytocin treated rats persisted. On day 10, the oxytocin-treated SHR males again had a significantly lower systolic blood pressure (p < 0.05). In female SHR, the same treatment with oxytocin affected neither blood pressure nor heart rate. These results show that oxytocin may cause a sustained decrease in blood pressure, without affecting heart rate, in male but not in female SHR. PMID- 9334990 TI - The recruitment pattern of single vasoconstrictor neurons in human. AB - The aim of this study is to determine the recruitment pattern among individual vasoconstrictor neurons under the baroreceptor-mediated influence in man. Spikes of single vasoconstrictor units were detected from microneurograms with a template-matching method. A total of 39 single vasoconstrictor units were detected. Single vasoconstrictor units were different from each other in their susceptibility to be activated in response to changes in the R-R interval or blood pressure. The units with higher firing probability had a shorter threshold R-R interval and a higher threshold diastolic blood pressure than units with lower firing probability. In sympathetic responses consisting of only one spike (single-spike responses), units with a lower threshold frequently appeared and units with a higher threshold joined mull-spike responses. The units with a short threshold R-R interval tended to have a long inhibitory latency from R wave, suggesting low conduction velocity. The correlation between firing probability and firing threshold and that between appearance in single-spike response and multi-spike response suggest a hierarchical manner of recruitment of vasoconstrictor units. For beat-to-beat responses, however, some deviation from the hierarchical recruitment was also observed. PMID- 9334989 TI - Effect of ibotenate lesions of the ventromedial hypothalamus on the water and salt intake induced by activation of the median preoptic nucleus in sodium depleted rats. AB - In this study we investigated the influence of a ventromedial hypothalamus (VMH) lesion with ibotenic acid on water and sodium intake and pressor responses induced by combined treatment of the median preoptic nucleus (MnPO) with angiotensin II (ANG II) and adrenergic agonists (phenylephrine, norepinephrine, isoproterenol and clonidine). Male Holtzman rats with a stainless steel cannula implanted into the MnPO and bilateral sham (vehicle) or VMH lesions with ibotenic acid were used. The ingestion of water and sodium and mean arterial pressure (MAP) were determined in separate groups submitted to sodium depletion with the diuretic furosemide (20 mg/rat). ANG II (10 pmol) injection into the MnPO of sham lesioned rats induced water and sodium intake and pressor responses. VMH-lesion reduced ANG II-induced water intake and increased saline intake. In sham rats phenylephrine (80 nmol) into MnPO increased, whereas norepinephrine (80 nmol) and clonidine (40 nmol) reduced ANG II-induced water intake while sodium intake was reduced only by clonidine into MnPO. In VMH-lesioned rats, phenylephrine reduced, noradrenaline increased and clonidine produced no effect on ANG II-induced water intake. In lesioned rats ANG II-induced sodium intake was reduced by phenylephrine and noradrenaline, whereas clonidine produced no change. ANG II induced pressor response was reduced in VMH-lesioned rats, but the pressor response combining ANG II and phenylephrine or noradrenaline in VMH-lesioned rats was bigger than sham rats. These results show that the VMH is important for the changes in water and sodium intake and cardiovascular responses induced by angiotensinergic and adrenergic activation of the MnPO. PMID- 9334991 TI - Nitric oxide synthase- and neuropeptide Y-containing subpopulations of sympathetic neurons in the coeliac ganglion of the Atlantic cod, Gadus morhua, revealed by immunohistochemistry and retrograde tracing from the stomach. AB - In this study retrograde tracing was used to locate sympathetic ganglion cells innervating the stomach of a teleost fish, Gadus morhua. A subpopulation of small neurons in the coeliac ganglion was retrogradely labelled after Fast Blue injection in the stomach wall. Neurons projecting to the myenteric plexus and muscle layers contained tyrosine hydroxylase immunoreactivity, and neurons projecting to submucosal layers and blood vessels contained neuropeptide Y immunoreactivity in addition to being tyrosine hydroxylase immunoreactive. A population of nitric oxide synthase containing tyrosine hydroxylase immunoreactive neurons was also found in the coeliac ganglion. These neurons were not frequently labelled after injection in any layer of the stomach. The presence of entero-enteric pathways was also surveyed, but too few enteric neurons were labelled with Fast Blue after injection in the coeliac ganglion to indicate a presence of an entero-enteric reflex. We conclude that in teleost fish, as previously reported in a variety of mammals, a pattern of target specific chemical coding of sympathetic neurons exists, but that all reflex systems of mammalian vertebrates are perhaps not present in fish. PMID- 9334992 TI - Selective autonomic and sensory deficits in slow transit constipation. AB - Chronic idiopathic constipation is likely to be a heterogeneous condition. Our previous studies on the stimulated sweating response suggested that autonomic dysfunction may be a cause in a subset of patients. Our aims were to test selectively the neural and sweat gland components of the sweat response and to test unmyelinated sensory fibres so as to determine whether a small fibre neuropathy is present. Twelve female patients with proven slow transit constipation and nineteen age-matched healthy volunteers took part in the study. The sensory tests included thermal thresholds and axon reflex vasodilatation in response to intradermal capsaicin, measured with a laser Doppler. Direct and axon reflex sweating was induced with intradermal methacholine and nicotine, respectively, and measured with an evaporimeter. Non-parametric tests were used for statistical comparison with a group of seven control subjects. Results are expressed as medians and range. All four patients who reported constipation from childhood had a selective deficit of unmyelinated afferent fibre function in the feet, with markedly elevated thresholds to warm sensation (controls 5.2; 4.3 10.6, patients 13.8; 11.8-16.1 delta T (degree C), P < 0.02) and heat pain (controls 10.6; 8.2-14.7, patients 18.1; 13.9-22.6 delta T (degree C), P < 0.05) and a reduced response to capsaicin (controls 47.0; 24-117, patients 13.5; 12-30 delta Flux (V), P < 0.005). In contrast, patients with adult onset constipation (n = 7) had a selective neural sweating deficit (controls 49.8; 32.0-61.8; patients 27.7; 7.3-44.3 g/m2 h, P < 0.05), indicating dysfunction of post ganglionic sympathetic cholinergic fibres. Patients from both groups were shown to have normally functioning sweat glands in direct response to methacholine. Our findings suggest that patients with severe chronic idiopathic constipation may have selective small fibre neuropathies, of which constipation is a manifestation. PMID- 9334993 TI - Role of spinal GABA receptors in depressor responses to chemical stimulation of the A5 area in normal and hypertensive rats. AB - Chemical stimulation of neurons in the pontine A5 area by microinjection of L glutamate lowers arterial blood pressure. The mechanism of this 'A5 depressor response' is not well-established. Here, we examine the involvement of spinal cord gamma-aminobutyric acid (GABA) receptors in this depressor response in normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Experiments were conducted in male WKY and age-matched SHR anaesthetised with sodium pentobarbitone and chloral hydrate. An intrathecal catheter was implanted with the tip located between T9 and L2. Three days later, rats were re anaesthetised and 10 nl of 40 mM L-glutamate was injected into the A5 area before, during and after, blockade of spinal cord GABA-A receptors by intrathecal injection of bicuculline methiodide (1 mM in 10 microliters phosphate-buffered saline). Injection of L-glutamate (10, 20, 40, 80 mM in 10 nl) produced depressor responses that were similar in WKY (n = 6) and SHR (n = 6). Intrathecal injection of bicuculline elicited a pressor response that was greater in SHR (n = 7, 28.5 +/- 7.6% increase in mean arterial pressure) than WKY (n = 11, 11.6 +/- 3.6%, p < 0.05). After bicuculline, the depressor response to injection of L-glutamate into the A5 area was eliminated in both WKY (n = 7) and SHR (n = 6). Intrathecal injection of vehicle had no effect on either resting arterial blood pressure or the depressor response to A5 stimulation. Basal blood pressure and control responses to A5 stimulation were fully restored by around 90 min after bicuculline injection in each animal. In separate groups of rats, intrathecal injection of muscimol elicited depressor responses that were greater in SHR (n = 6, -32.0 +/- 6.2%) than WKY (n = 6, -17.3 +/- 1.5%, p < 0.05). Our results suggest that the A5 depressor response is due to a projection from the A5 area to the spinal cord. This projection acts directly, or through a spinal interneuron and uses GABA as a neurotransmitter. Furthermore, our results indicate a hyper responsiveness to GABA-A receptor stimulation in SHR since intrathecal bicuculline elicited much greater increases and intrathecal muscimol elicited much greater decreases, in blood pressure in SHR than in WKY. Finally, it seems likely that the A5-spinal depressor pathway is less effective in SHR than WKY under physiological conditions since chemical stimulation of the A5 area with L glutamate produced a comparable depressor response in both strains. PMID- 9334994 TI - Heart rate variability in workers exposed to carbon disulfide. AB - It is assumed that the cardiovascular impairments resulting from CS2 exposure may be associated with some functional disturbances within the autonomic nervous system. We adopted the heart rate variability (HRV) analysis to investigate the sympathetic and parasympathetic functions of the autonomic nervous system in workers exposed to carbon disulfide. The studies were performed on 152 workers, aged 24-66, with the period of exposure ranging from 5-38 years and 93 age matched, non-exposed, healthy individuals as the control group. The HRV analysis concerned time-domain (AVG R-R, SD R-R, modal, median, minimum and maximum values) and frequency-domain indices (power spectrum in the very low-VLF, low-LF and high-HF frequency bands) calculated using the fast Fourier transformation. In the exposed group, neurovegetative regulation impairments could be observed. They were expressed as increased heart rate at rest, reduced power spectrum: total (TPS) and within HF, LF and VLF frequency bands as well as the absence of the physiological dependence of HRV parameters on age. These abnormalities could be found even in the group of workers exposed to the lowest CS2 levels (0-10 mg/m3) and they concerned the VLF band. In workers under conditions of exposure to 10-18 mg/m3 the dysfunction of the autonomic control referred both to its sympathetic and parasympathetic part and was found to be more intense in workers exposed to the highest CS2 concentrations (over 18 mg/m3). In view of our findings we concluded that occupational exposure to carbon disulfide may bring about an impaired neurovegetative regulation of the cardiovascular function. PMID- 9334995 TI - Reflex control of autonomic function induced by posture change during the menstrual cycle. AB - This study was designed to determine whether autonomic regulation induced by posture changes varies during the menstrual cycle. Heart rate variability (HRV) was analyzed in women with normal menstrual cycles (n = 10, age range 20-37 years) during 3 min periods of controlled frequency breathing (15 breaths/min) in supine followed by sitting positions. In a supine or sitting position, high frequency (HF) components of HRV, which reflects only parasympathetic activity, were significantly high in the follicular phase compared with those in the menstrual phase, suggesting that parasympathetic nerve activity increases in this phase. Following the change of position to sitting from supine, the HF component decreased significantly in the menstrual, ovulatory and luteal phases, but not the follicular or premenstrual phase. After changing the position to sitting, the low to high frequency component ratio, which reflects the balance of autonomic nerve activities, was increased significantly in the menstrual, luteal and premenstrual phases, indicating that sympathetic nerve activities in these phases became predominant by the sitting position. These results suggest that parasympathetic nerve activity is predominant in the follicular phase, resulting in an impairment of baroreflex caused by posture changes. Moreover, baroreflex control of the sympathetic component, not the parasympathetic component, increases in the premenstrual phase, while the reflex response of the sympathetic component is less in the ovulatory phase compared with the menstrual or luteal phase. We concluded that baroreflex regulation of autonomic functions induced by changing positions is modified during the menstrual cycle. A difference of a balance of ovarian hormones may be responsible for these changes of autonomic functions during the menstrual cycle. PMID- 9334996 TI - Selective innervation of different target tissues in guinea-pig cranial exocrine glands by sub-populations of parasympathetic and sympathetic neurons. AB - This study has used multiple-labelling immunohistochemistry and quantitative analysis to examine the projections of subpopulations of parasympathetic and sympathetic neurons to different vascular and secretory structures in five cranial exocrine glands of guinea-pigs. Multiple subpopulations of parasympathetic axons, identified by immunoreactivity (IR) for various combinations of peptides, innervated arteries, arterioles, ducts and acini in sublingual, submandibular, parotid, lacrimal and zygomatic glands, although axons were absent from ducts in the parotid gland. Most parasympathetic axons contained IR for vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY), with or without enkephalin (Enk). The proportion of parasympathetic axons that contained Enk-IR varied greatly between target tissues and glands: Enk-IR was more common in axons supplying secretory ducts, acini and arterioles than in axons innervating more proximal arteries; Enk-IR was less common in axons supplying the lacrimal gland than axons supplying the submandibular, lacrimal and zygomatic glands. Sympathetic axons with IR for tyrosine hydroxylase (TH) innervated arterial vessels in all glands, but innervated secretory structures only in the salivary glands. Sympathetic axons supplying proximal arterial segments often contained NPY-IR and sometimes also contained IR for dynorphin. Dynorphin-IR was more common in axons in the parotid, lacrimal and zygomatic glands than in the sublingual and submandibular glands. In contrast, axons supplying arterioles, ducts and acini lacked peptide IR. These results indicate that neuronal pathways regulating proximal arteries in cranial exocrine glands are different from the neuronal pathways regulating arterioles and acini, and may be different from neurons projecting to proximal secretory ducts. Furthermore, the peptides enkephalin, NPY and dynorphin are likely to make variable contributions to autonomic neurotransmission in different arterial segments and in different cranial exocrine glands. PMID- 9334997 TI - Angiotensin converting enzyme inhibition improves baroreflex-induced noradrenaline spillover responses in rabbits with heart failure. AB - Impaired baroreflex function is a characteristic feature of congestive heart failure (CHF), although the mechanism is obscure. This study examined the hypothesis that activation of the renin-angiotensin system contributes to baroreflex dysfunction in CHF. The acute effects of an angiotensin converting enzyme inhibitor, enalaprilat, on baroreflex-mediated changes in heart rate (HR), total and renal noradrenaline (NA) spillover rates were examined in conscious rabbits with doxorubicin-induced cardiomyopathic CHF. Studies were performed under resting conditions and in response to changes in mean arterial pressure (MAP) induced by sodium nitroprusside and phenylephrine infusions. Seven saline treated (normal group) and 11 doxorubicin-treated rabbits (1 mg/kg administered intravenously twice weekly) were studied after 4 and 6 weeks' treatment. Five CHF rabbits received saline (C group) and 6 enalaprilat infusion (ACEI group) during each study period. After 4 weeks of doxorubicin, baroreflex-HR responses were normal, whereas baroreflex-NA spillover responses were enhanced. Enalaprilat infusion shifted the HR-MAP curve downwards to the left but had no effect on the NA spillover-MAP curves. After 6 weeks of doxorubicin, when CHF was established, baroreflex-HR and NA spillover curves were depressed. At this stage, enalaprilat had little effect on the HR-MAP curve but restored towards normal the NA spillover-MAP curves. The results suggest that the endogenous renin-angiotensin system contributes to attenuated baroreflex responses when CHF is established. PMID- 9334998 TI - Contralateral projections by preganglionic neurons to the sympathetic trunk of the puffer fish, Takifugu niphobles. AB - Previous studies have shown that the sympathetic preganglionic neurons of teleosts send axons to the sympathetic trunk on the contralateral side. After severing the spinal nerve roots at a level proximal to the sympathetic ganglia (i.e., nerve roots containing the preganglionic axons) on one side of puffer fish, Takifugu niphobles, horseradish peroxidase was applied to the other side of the sympathetic trunk. Retrogradely labeled sympathetic preganglionic neurons were found bilaterally in the central autonomic nucleus (a distinct cell column in the rostral part of the spinal cord). The contralaterally labeled neurons were located almost exclusively in the caudal part of the nucleus. These results suggest that some sympathetic preganglionic neurons in teleosts, unlike those in other vertebrates, send their axons across the midline to the contralateral nerve roots. PMID- 9334999 TI - Responses of distinct types of sympathetic neuron to stimulation of the superior laryngeal nerve in the cat. AB - Stimulation of afferents in the superior laryngeal nerve (SLN) leads to apnea and evokes reflexes in sympathetic neurons. It is not clear whether these reflexes are secondary to changes in the brainstem respiratory network or due directly to the afferent input on neurons belonging to central sympathetic pathways. To clarify this question, single thoracic preganglionic sympathetic neurons projecting into the cervical sympathetic trunk (CST) were classified as described previously and then tested for their responses to electrical stimulation of the superior laryngeal nerve (SLN) in chloralose-anesthetized, paralysed and artificially ventilated cats. SLN stimulation was performed with intensities sufficient to suppress central inspiratory activity detected by phrenic and recurrent laryngeal nerve recordings. Sympathetic neurons were tested under different levels of respiratory drive. Thirteen group I (putative muscle vasoconstrictor) neurons were mostly activated by SLN stimulation when respiratory drive was low, but depressed when it was high; this was due to the change in inspiration-related activity. Ten of eleven group II (putative cutaneous vasoconstrictor) neurons were depressed during SLN stimulation. This inhibition was independent of central respiratory drive. Inhibition also occurred in those neurons which predominantly discharged during postinspiration. Eight group III neurons which showed a discharge confined to inspiration were inhibited but mostly not silenced by SLN stimulation. Group IV (functionally unclassified) neurons either showed no response (n = 5) or were slightly inhibited (n = 2). The responses of group I neurons, but not the responses of group II and group III neurons, showed a significant positive correlation with those of systemic blood pressure. The observed responses corroborate the classification made previously. The results also demonstrate that the responses of sympathetic neurons to SLN stimulation are not merely due to the respiratory modulation of their activity, but rather consist of two components, one occurring independently of and the other secondary to, the changes in respiration. PMID- 9335000 TI - Activation of 5-HT1P receptors on submucosal afferents subsequently triggers VIP neurons and chloride secretion in the guinea-pig colon. AB - The role of vasoactive intestinal peptide (VIP) was investigated when mucosal stroking and 5-hydroxytryptamine (5-HT) were used to activate neural reflexes that stimulate chloride secretion in the guinea pig colon. Muscle-stripped segments of colon containing intact submucosal ganglia without myenteric ganglia were set up in modified flux chambers in order to record short-circuit current (Isc). Mucosal stroking with a brush for 1 s or a pulse of 5-HT (injection of 15 microliters of 100 microM 5-HT into 1.5 ml of mucosal solution) caused an increase in Isc that was reduced by the VIP antagonist, neurotensin6-11-VIP7-28, in a concentration-dependent manner. The Isc responses to mucosal stroking and a 5-HT pulse were reduced by 53% and 58%, respectively, by 2 microM neurotensin6-11 VIP7-28. The residual Isc response in the presence of neurotensin6-11-VIP7-28 was abolished by atropine. Blockade of 5-HT1P receptors on submucosal afferent neurons decreased Isc responses to stroking or a 5-HT pulse. The residual Isc response after 5-HT1P receptors were blocked was reduced by only 11-14% by neurotensin6-11-VIP7-28. In the presence of blockade of both 5-HT1P and VIP receptors, atropine abolished the Isc response to both stimuli. The observations suggest that the neural circuitry activated by stroking includes at least two independent pathways. One pathway contains VIP neurons which receive inputs directly or indirectly from 5-HT1P receptor-containing afferents. A second pathway involves muscarinic cholinergic transmission that is independent of 5 HT1P and VIP receptor activation. PMID- 9335001 TI - Cerebral cortex regional blood flow and tissue oxygen tension during the trigeminal depressor response in rabbits. AB - The trigeminal depressor response (TDR) is characterized with profound hypotension caused by sympathetic outflow decreases. The purpose of this study was: (1) to compare the hemodynamic changes during acute hypotension induced by the TDR with those induced by electrical stimulation of the vagal nerve (VS) as models of sympathetic inhibition and vagal activation and (2) to investigate the effects of nitrous oxide (N2O), a well-known sympathomimetic, on the hemodynamic changes during the TDR. Male Japan White rabbits were anesthetized with halothane in oxygen and mechanically ventilated. The electrode pair for the TDR was inserted into the submucosal tissue of the animal's tongue. The pair for the VS was applied to the isolated left vagal trunk. Both the TDR and the VS produced similar decreases in mean arterial pressure (MAP) and cerebral cortex regional blood flow (rCBF), although the decreases in heart rate (HR) and aortic blood flow were greater during the VS. Cerebral cortex tissue oxygen tension in both groups decreased slightly. 50% N2O, producing MAP elevation and HR decrease, ameliorated the hemodynamic changes including rCBF reduction during the TDR. A sudden diminution of sympathetic tone following noxious stimulation of the orofacial area, as in the case of the TDR, may be a possible trigger mechanism for vasodepressor reactions in dental patients. The sympathomimetic and possible antinociceptive effects of N2O may explain, at least in part, the preventive effects on vasodepressor reactions during dental procedures. PMID- 9335002 TI - Synapse alteration in hippocampal CA3 field following entorhinal cortex lesion. AB - To model one aspect of the neurodegeneration observed in Alzheimer's disease and to investigate the synaptic alteration of the hippocampus associated with entorhinal cortex lesion, ibotenic acid was used to produce selective unilateral neuronal loss in rat entorhinal cortex. Immunohistological and microdensitometrical analyses confirmed ibotenic acid lesion of the entorhinal cortex after 3 months and showed a decrease of synaptophysin-immunoreactive substances in the stratum lucidum of the CA3 field. This study demonstrates that entorhinal cortex lesion can lead to synaptic alterations and cause damage to presynaptic terminals with projecting area in the disruption of the entorhinal cortex hippocampus relay passage. PMID- 9335003 TI - S100 beta prevents the death of motor neurons in newborn rats after sciatic nerve section. AB - We have examined whether S100 beta rescues axotomized spinal motor neuron death. Animals that had undergone transection of the sciatic nerve at birth were treated either with S100 beta, or the vehicle. The number of surviving motor neurons and the motor neuron diameter was assessed 14 days later. Treatment with S100 beta rescued motor neuron death and preserved the motor neuron diameter in the lesioned side. These results suggest that S100 beta is a neurotrophic factor for motor neurons in vivo and this agent may have therapeutic potential in damaged motor neuron disorder. PMID- 9335004 TI - Schwann cell extracellular matrix protein production is modulated by Mycobacterium leprae and macrophage secretory products. AB - Extracellular matrix (ECM) protein deposition is an important feature of leprous nerves, where Schwann cells (SCs) and macrophages are the main hosts for Mycobacterium leprae. Since, SCs are involved in the synthesis of ECM proteins and its production is regulated by macrophage secretory factors, the present study aimed to determine in vitro, the effect of M. leprae infection and macrophage secretory products on secretion of ECM proteins by SCs in two strains of mice, Swiss White (SW) and C57BL/6, that are known to differ in their nerve pathology and macrophage functions in response to infection. Following six days of M. leprae infection, SCs from SW mice responded with increased secretion of 14C-leucine radiolabelled proteins and a concomitant increase in laminin and collagens type I, III and IV, as determined by enzyme-linked immunosorbent assay. In contrast infected C57BL/6 SCs responded with decreased secretion of total proteins and fibronectin. Exposure of SCs to macrophage conditioned medium resulted in decreased ECM protein secretion in both strains of mice. This decrease was a function of protein breakdown by macrophage derived proteases and also active regulation by macrophage secreted cytokines. A similar effect of M. leprae and macrophage secretory products on SC metabolism in leprous nerves would have major ramifications on damage and repair activities. In addition ECM proteins would also influence the composition of the infiltrating cell population in lepromatous and tuberculoid nerves. PMID- 9335005 TI - Chorea-acanthocytosis with polyclonal antibodies to ganglioside GM1. AB - A patient with chorea-acanthocytosis presenting with axonal neuropathy showed an elevation in IgM polyclonal antibodies to the GM1 ganglioside, which were estimated by enzyme-linked immunosorbent assay and complement-mediated liposome immune lysis assay (LILA). This is the first demonstration of such antibodies in chorea-acanthocytosis. Anti-GM1 antibodies might have directly caused the axonal neuropathy by binding to GM1 or cross-reactive antigens in the nerves. PMID- 9335006 TI - Congenital muscular dystrophy with partial deficiency of merosin. AB - We present a Japanese patient who has congenital muscular dystrophy, with partial merosin deficiency. The patient had characteristic findings of clinical features and brain MRI. Muscle biopsy showed advanced muscular dystrophy, with greatly reduced muscle fibers and massive infiltration of interstitial connective and fatty tissues. On immunostaining for cytoskeletal proteins, merosin was greatly reduced. The other cytoskeletal proteins, including dystrophin and 50 kDa alpha sarcoglycan were normally expressed around all muscle fibers. PMID- 9335007 TI - A longitudinal study of circulating lymphocyte subsets in the peripheral blood during the acute stage of Guillain-Barre syndrome. AB - Activated T cells are implicated in the pathogenesis of Guillain-Barre syndrome (GBS). Blood samples from 16 patients with GBS were studied with flow cytometry during the acute stage of their disease to define circulating lymphocyte populations. During the progressive phase of GBS the T-suppressor/inducer (CD4/CD45RA) subset was decreased (10.3 +/- 4.4%, P < 0.05) and the T-helper/ inducer (CD4/CD29) subset was increased (34.9 +/- 9.2%, P < 0.05) compared to sex and age matched patients with other neurological diseases (OND, 15.5 +/- 5.7% and 27.8 +/- 8.6%, respectively) and healthy controls (16.5 +/- 6% and 28.1 +/- 8.5%, resp.). Within the CD8 population, the activated T-cytotoxic/suppressor (CD8/CD38) subset was increased during the progressive (13 +/- 10.1%, P < 0.05) and plateau phase (16.4 +/- 16.9%, P < 0.01) of GBS compared to OND patients (6.2 +/- 2.3%) and healthy controls (5.8 +/- 2.5%). The proportion of activated T cells (CD3/CD25) was increased during the progressive (9.3 +/- 3.8%, P < 0.05) and plateau phase (11.5 +/- 5.5%, P < 0.01) compared to OND patients (6.6 +/- 2.7%) and healthy controls (5.5 +/- 2.5%). The changes of T cell subsets normalized during the early recovery phase of GBS. 2 patients with serological evidence of antecedent cytomegalovirus (CMV) infection had abnormal high proportions (mean +/- 2 (SD) of healthy controls) of CD8 lymphocytes and correspondingly abnormal low proportions of CD4 lymphocytes during all phases of GBS. In contrast, the CD8 proportions were abnormal low in 3 patients with serological evidence of recent Campylobacter jejuni infection. There was no correlation between the proportions of lymphocyte subsets and the disability score during the maximum of the disease and after half a year. In conclusion, we found further evidence of T cell activation during the acute stage of GBS by the demonstration of an increased proportion of activated CD8+ T cells, which may be directly cytotoxic to Schwann cells. The abnormalities of the CD8 subset in some GBS patients seem to depend on the nature of the preceding infection. PMID- 9335008 TI - Plasma superoxide dismutase and glutathione peroxidase activity in sporadic amyotrophic lateral sclerosis. AB - To determine the possible role of oxydative stress in the pathology of amyotrophic lateral sclerosis (SALS), we measured the plasma activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX), together with GPX and malone dialdehyde (MDA, a marker of lipoperoxydation) plasma concentrations in a sample of 21 SALS patients and 7 normal control (NC) subjects. MDA concentration and SOD activity were significantly higher, whereas GPX activity was significantly lower in SALS patients than in NC. Increased MDA concentration provides indirect confirmation of excess lipoperoxydation. Increased plasma SOD activity might reflect the involvement of extra-cellular SOD (SOD3), a hitherto unreported finding in SALS. Impaired GPX activity, which has already been found in red blood cells and brain tissue of SALS patients, might play a part in the pathogenesis of this disease. PMID- 9335009 TI - Identification and characterization of an anti-glial fibrillary acidic protein antibody with a unique specificity in a demented patient with an autoimmune disorder. AB - We detected an antibody to a 48 kd antigen of the central nervous system in the serum from a demented patient with an autoimmune disorder. To identify and characterize the antigen, we screened a human cerebral cDNA library and performed immunoblot analysis following two-dimensional gel electrophoresis (2-D blotting). The sequences of the isolated cDNA fragments were homologous to human glial fibrillary acidic protein (GFAP). Two-D blotting using patient serum revealed that the antibody reacted with a restricted subset of GFAP molecules which exhibited relatively high isoelectric points. Furthermore, to elucidate the importance of the anti-GFAP antibody in dementia, we screened for the presence of an anti-GFAP antibody in the sera of 46 demented patients: 26 with Alzheimer's disease and 20 with vascular dementia (VD). We found an anti-GFAP antibody in the serum of only one patient with VD. Two-D blotting revealed that the anti-GFAP antibody in the serum from the VD patient reacted with a more acidic subset of GFAP molecules compared with the anti-GFAP antibody from our patient. In conjunction with the fact that the GFAP molecule with high isoelectric point was insoluble and less degraded, these results suggested that the anti-GFAP antibody in the serum of our patient was not generated due to a secondary response to soluble and degraded GFAP which leaked through the damaged blood-brain barrier as found in the VD patient, but was generated actively on the basis of dysregulation of the immune system. Possible effects of the autoantibody on astrocytic function and the pathogenesis in dementia are discussed. PMID- 9335010 TI - Altered tumor growth factor beta mRNA expression is associated with thymectomy related clinical remission in myasthenia gravis. AB - Clinical remission in myasthenia gravis after thymectomy and immune suppressive treatment may be reflected by changes of immunoregulatory cytokines. Mononuclear cells from paired blood samples from fifteen patients with myasthenia gravis before and after thymectomy and immunosuppressive therapy were examined by in situ hybridization for acetylcholine receptor-induced mRNA expression of the T helper type 1 proinflammatory cytokines interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha) and TNF-beta, the T helper type 2 cytokines interleukin 4 (IL-4) and IL-10 and the immune response downregulating cytokine tumor growth factor beta (TGF-beta). A significant change was observed for TGF beta, with increased expression after treatment and induction of clinical remission. Augmentation of TGF-beta may play a role in the induction of remission in myasthenia gravis. PMID- 9335012 TI - Autosomal dominant spastic paraplegia linked to chromosome 2p: clinical and genetic studies of a large Japanese pedigree. AB - Autosomal dominant spastic paraplegia (ADSP) is a genetically heterogenous disorder. To date, 3 loci of ADSP have been identified on chromosome 2p, 14q, and 15q, but specific gene mutations remain unknown. To determine the genetic background of ADSP in the Japanese, we studied a large 3-generation pedigree, clinically and genetically. Of the 36 individuals clinically examined, 15 were affected. The main feature in the affected individuals was a slowly progressive spastic paraplegia, associated with upper limb hyperreflexia (58%), reduction of vibration sense (27%) and bladder disturbance (13%). Age at onset ranged from 13 to 50 years with a mean of 30.3 +/- 14.2 (SD). There were 6 parent-child pairs with anticipation and at least 3 others with 'anti-anticipation'. Linkage with 14q and 15q ADSP loci was excluded, and a highly significant lod score was obtained only in the case of the 2p locus (Zmax = 3.53 for D2S400/D2S352, at theta = 0.00). Our study is the first to confirm the existence of 2p-linked ADSP in the Japanese. There is a significant variety in age at onset and disease severity in these 2p-linked families, but the implication for underlying ADSP mutation is not clear. PMID- 9335011 TI - Progressive dystonia with optic atrophy in a Jewish-Iraqi family. AB - The combination of progressive dystonia and optic atrophy is extremely rare and its morphological, metabolic and genetic basis is unknown. In a family of 9 children (8 males) born to consanguineous Israeli-Jewish-Iraqi parents, we identified four brothers who developed the syndrome at the end of the first decade. Patients had hemi or bilateral dystonia associated with striatal, mainly putaminal, atrophy on CT and MRI, various degrees of optic atrophy, minimal corticospinal tract involvement, normal intelligence and no peripheral nervous system or systemic abnormalities. No causative metabolic defect was identified. None of the several known mitochondrial DNA mutations associated with Leber's hereditary optic neuropathy (LHON) or with LHON with dystonia were detected. Likewise, linkage to the idiopathic torsion dystonia region on chromosome 9q34 was excluded. It is suggested that this in our patients might be due to a yet unidentified genomic, autosomal recessive mutation. PMID- 9335013 TI - Dissociation in the neural control of single-joint and multi-joint movements in the thalamic ataxia syndrome. AB - We report a patient presenting with a right thalamic ataxia syndrome following a hemorrhage located in the left lateral and posterior thalamus. We investigated the fast goal-directed movements of the wrists (single-joint movements) and the fast pointing movements in the upper limbs (multi-joint movements). On the right side, single-joint movements were markedly hypermetric and characterized by an asymmetry in kinematics, an abnormality of ballistic movements which is considered to be a fundamental cerebellar disorder. By contrast, rapid multi joint movements were only very slightly impaired. These results suggest that ballistic movements of the wrist are under the strong influence of the cerebello thalamo-cortical pathway, while rapid pointing multi-joint movements in upper limb are mostly influenced by another pathway emerging from the lateral cerebellum, possibly the dentato-rubral or the dentato-reticular projections in the brainstem. The roles of these neuroanatomical pathways in the control of fast single-joint and multi-joint movements are discussed. PMID- 9335014 TI - Magnetization transfer ratio changes in a symptomatic lesion of a patient at presentation with possible multiple sclerosis. AB - Short-term changes of magnetization transfer ratio (MTR) of a symptomatic lesion in a patient at presentation with clinically isolated syndrome suggestive of multiple sclerosis (MS) are presented. These changes strictly correlated with the evolution of the corresponding clinical symptomatology. This suggests that following changes of individual strategically located lesions might be useful to monitor evolution of demyelinating white matter diseases. PMID- 9335015 TI - Correlation between neuromorphometry in the substantia nigra and clinical features in Parkinson's disease using disector counts. AB - Previous studies based on single sections have suggested a significant correlation between pigmented neuronal loss in the substantia nigra (SN) and clinical features in Parkinson's disease (PD). However, disector (DS) counts unbiased and accurate stereological estimates have not been available. To evaluate total neuron numbers in the pars compacta of the substantia nigra (SNpc) in relation to clinical features, we estimated the neuron counts in the SNpc by the DS method in brain samples from 12 controls and 12 PD patients. The total number of pigmented neurons in the whole SNpc was significantly reduced in PD patients (to 45% of the control mean, P < 0.001). The density of pigmented neurons (neuron/mm3) was reduced to 51% of the average control value (P < 0.001). No significant difference was seen in the volume (mm3) of the SNpc between PD patients and controls. Furthermore, the total number of pigmented neurons in the SNpc showed a significant negative correlation with the duration of disease (r = 0.86, P < 0.001) and with the stage of disease (r = -0.58, P < 0.05) in PD patients. Using an unbiased neuron counting method, these relationships, for the first time, demonstrate that the more severe pigmented neuronal loss in the SNpc is associated with the longer duration and the more severe stage of disease in PD patients. PMID- 9335016 TI - Decrease of neurons in the medullary arcuate nucleus of multiple system atrophy: quantitative comparison with Parkinson's disease and amyotrophic lateral sclerosis. AB - The physiological functions of the medullary arcuate nucleus are supposed to be involved in autonomic cardioventilatory regulation, but neuropathological studies on neurodegenerative diseases have rarely reported about the arcuate nucleus. We quantitatively examined the neuronal density of the arcuate nucleus in patients with multiple system atrophy (MSA, n = 3), Parkinson's disease (PD, n = 3), amyotrophic lateral sclerosis (ALS, n = 2), and control subjects (n = 6), and statistically compared the findings in each group. Although the neuronal densities in PD and ALS patients were not different from that in the controls, MSA patients showed a marked depletion of neurons in the arcuate nucleus. The neuronal density (/mm2, mean +/- SEM) in the arcuate nucleus was 9.27 +/- 10.4 in MSA, and was significantly decreased (P < 0.05; Wilcoxon test), compared with that in control subjects (87.1 +/- 12.2). These results suggest that the lesioned arcuate nucleus is related to the pathogenesis of dysatonomia in MSA. PMID- 9335017 TI - Serial MR findings of chronic idiopathic ataxic neuropathy. AB - We report the case of a 70-year-old woman with chronic idiopathic ataxic neuropathy. Neurological examination, electrophysiological studies and a sural nerve biopsy suggested involvement of the axon and the dorsal root ganglia. Laboratory examination showed polyclonal elevation of serum IgA. MRI showed high signal intensity in the posterior column of the lumbar spinal cord on T2-weighted images. High signal intensity in the posterior column of the cervical spinal cord on T2-weighted images appeared during the course of the illness. MR abnormalities in this case may reflect the degeneration of the posterior column of the spinal cord subsequent to sensory ganglionic neuropathy. PMID- 9335018 TI - Successful application of pentoxifylline in the treatment of HTLV-I associated myelopathy. AB - Fifteen patients with human T-cell lymphotropic virus type-I (HTLV-I)-associated myelopathy (HAM) were treated in an uncontrolled preliminary trial by oral administration of pentoxifylline (PTX). Motor function, neurological evaluation, immunological markers and parameters were evaluated after four weeks. In 13 of the 15 patients, motor disability, especially spasticity, improved substantially. PTX suppressed spontaneous proliferation of peripheral blood mononuclear cells in 14 of the 15 patients at four weeks. No adverse effect was observed. We concluded that PTX may be a safe and beneficial agent for the treatment of HAM. PMID- 9335019 TI - Painful small-fibre multifocal mononeuropathy and local myositis following influenza B infection. AB - A 47-year-old man experienced multifocal mononeuropathy and putative ganglionopathy associated with influenza B infection, characterized by aching and dysesthesia in the right arm and left leg with normal deep sense. He displayed muscle atrophy in the affected limbs, which might have resulted from local myositis or a disorder similar to neuralgic amyotrophy. Sural nerve biopsy revealed a severe loss of unmyelinated and thinly myelinated fibres, consistent with a small fibre neuropathy, without evidence of angiopathy or inflammation. We could not detect any other cause of the neuropathy except influenza B. In this case, it may be inferred that small-diameter neurons in the dorsal root ganglia and thinly- or nonmyelinated fibres were selectively involved through a post infectious immune process. To our knowledge, small fibre neuropathy following influenza B has never been reported. PMID- 9335020 TI - Primary central nervous system leukemia with a novel chromosomal translocation. AB - A case of central nervous system (CNS) leukemia with normal bone marrow, associated with a novel chromosomal abnormality, is described. A 58 year-old woman complained of hearing disturbance, severe headache and vomiting, and showed signs of meningeal irritation, as well as papilledema and bilateral dysacusis. Immature atypical cells were found in the cerebrospinal fluid (CSF) with elevated pressure, pleocytosis, increased protein and decreased glucose levels. She was diagnosed as having neoplastic meningitis. In spite of intensive investigations, including bone marrow puncture, malignancies were not found in organs other than intra-cranial site. The symptoms and CSF findings were temporarily improved with chemotherapy and irradiation, but she relapsed into neoplastic meningitis. The anaplastic cells in CSF were positive with CD45 by immunocytochemistry, and were positive by peroxidase staining. Thus, the anaplastic cells were considered to be myelocytic leukemic cells. Chromosomal analysis showed that these leukemic cells had a novel chromosomal abnormality: 46XX, 4q+, 10q-, 16q-. There has been no report of leukemic meningitis without bone marrow abnormalities. It is possible that this peculiar abnormal chromosome is related to the primary infiltration of the central nervous system. With this novel chromosomal abnormality, this case is important for considering the mechanism of primary leukemic meningitis. PMID- 9335021 TI - Epileptic nystagmus in infancy. AB - Epileptic nystagmus (EN) is a rare form of nystagmus that occurs only during epileptic seizures. We report an infantile case in which EN was first noted at 10 days of age. Electronystagmography showed a right-beating nystagmus with predominantly linear slow phases that traversed the midline. Neuro-imaging revealed dysplasia of the left middle temporal gyrus extending posteriorly into the parieto-occipital cortex. The right hemisphere and subcortical structures appeared normal. Perfusion studies demonstrated interictal hypoperfusion with ictal hyperperfusion in the left temporal lobe. Electrocorticography demonstrated spiking over the left temporal-parieto-occipital region. Following extensive surgical resection of this area and weaning of anti-convulsants, the child has remained seizure-free without nystagmus. This case demonstrates the cortical origin of EN, and shows that infant cortex has functioning efferent connections to brainstem oculomotor centres from 10 days of age. PMID- 9335022 TI - Leber's 'plus'. PMID- 9335023 TI - How good are the results of ACL reconstruction? PMID- 9335024 TI - Arthroscopic-assisted anterior cruciate ligament reconstruction with the central third patellar tendon. A 5-8-year follow-up. AB - We reviewed 89 arthroscopically assisted patellar tendon anterior cruciate ligament (ACL) reconstructions for chronic isolated injuries with an average follow-up of 7 years (range 5.4 to 8.6 years). Pain was present in 7 knees (8%). Giving-way symptoms were reported by 7 patients (8%). A KT-2000 side-to-side difference over 5 mm at 30 lbs was recorded in 12 cases (16%). The pivot shift was glide in 17 cases (19%) and clunk in 10 (11%). A 3 degrees-5 degrees extension loss compared with the normal side was present in 20 knees (22%) and 6 degrees-10 degrees in 4 knees (4%). The intra-articular exit of the femoral tunnel was misplaced in the anterior 50% of the condyles along the roof of the notch in 10% of the knees. This positioning significantly (P = 0.003) increased the frequency of graft failure (62.5%) compared with the cases with a more posterior placement (graft failure 12%). An anterior position of the intra articular exit of the tibial tunnel (in the anterior 15% of the sagittal width of the tibia) significantly (P = 0.01) increased the frequency of extension loss > 5 degrees. Medial meniscectomy was associated with a 35% incidence of narrowing of the medial joint space, which was significantly higher compared with knees with normal menisci (9%; P = 0.04) or with medial meniscal repair (7%; P = 0.05). In conclusion this study showed satisfactory anterior stability (KT-2000 side-to side difference up to 5 mm and pivot absent or glide) in 83% of the knees. This percentage increases to 88% in the knees with a correct posterior and proximal femoral tunnel placement. Accuracy in tunnel positioning is essential for the success of ACL surgery. Meniscal repair was effective in decreasing joint space narrowing and should be attempted when possible. PMID- 9335025 TI - The anterior cruciate ligament does play a role in controlling axial rotation in the knee. AB - This study deals with the influence of peroperative ligament tension on total tibial rotation at different knee flexion angles. Fourteen human cadaver knees with a mean age of 56 years (range 42-84 years) were examined. The cadaver knees were subjected to internal/external (i/e) rotational torque of 6 Nm, at 10, 30, 50, 70 and 90 deg of knee flexion. The mean total i/e rotation with the anterior cruciate ligament (ACL) intact at 10 deg of knee flexion was 30.4 deg and after removing the ACL, 33.1 deg. At 10 and 30 deg of knee flexion, the increase in i/e rotation was significant, while there was no significant difference in mean values at greater knee flexion. Ligament reconstruction with a tension of 5 N at 30 deg of knee flexion using either the over the top or through the femoral condyle reconstructive procedure restored normal tibial rotation. With increased graft tension the knee motion was increasingly restricted at low angles of knee flexion. Our results indicate that the ACL does play a role in limiting axial rotation, and even minor tensioning forces introduced in any of the two ACL reconstructions used produced restricted knee motion. PMID- 9335026 TI - Anterior cruciate ligament (ACL)-deficient knee with degenerative arthrosis: treatment with an isolated autogenous patellar tendon ACL reconstruction. AB - We evaluated 58 patients (mean age 30.4 years) who had undergone an isolated anterior cruciate ligament (ACL) reconstruction for chronic instability (mean time from injury to surgery, 8.2 +/- 5.2 years) and showed radiographic evidence of degenerative arthrosis. Objective evaluation at a mean of 4.1 years postoperatively included KT-1000 arthrometer stability, range of motion, and quadriceps muscle strength testing. Subjective analysis at a mean of 5.5 years postoperatively included rating of pain, stability, activity level, and a total score both preoperative and postoperative. Patients were divided into two groups: group 1 (n = 28) with a follow-up < or = 5 years (mean 3.3 years); group 2 (n = 30) with a follow-up > 5 years (mean 7.2 years). Results were analyzed by length of follow-up and by the grade and compartment of arthrosis. All patients enjoyed a full range of motion preoperatively and postoperatively. The mean KT-1000 arthrometer manual maximum difference improved from a mean of 8.2 mm preoperatively to 2.4 mm postoperatively. All subjective scores showed statistically significant improvement over the preoperative values. Patients with medial compartment arthrosis reported a better subjective total score (mean 87) than patients with lateral compartment (mean 73) or bicompartmental (mean 79) arthrosis, but there was not a statistically significant difference. There was no correlation between pain, stability, or total scores and time after surgery. Patients in groups 1 and 2 had equal objective stability and similar subjective scores, but group 2 reported a lower activity level. An isolated ACL reconstruction can provide long-term stability and symptomatic pain relief in patients with chronic instability and arthrosis. The procedure has low morbidity and does not compromise future tibial osteotomy or total knee replacement. PMID- 9335027 TI - Is bracing after anterior cruciate ligament reconstruction necessary? A 2-year follow-up of 78 consecutive patients rehabilitated with or without a brace. AB - The aim of this study was to evaluate the effect of a standard postoperative rehabilitation knee brace on function, stability and postoperative complications at the 2-year follow-up after anterior cruciate ligament (ACL) reconstructive surgery. Seventy-eight consecutive patients with a unilateral chronic ACL rupture reconstructed by the same surgeon using the endoscopic "all-inside" technique, patellar tendon autograft and interference screw fixation were included in the study. The rehabilitation followed a standard protocol. Group A included 39 patients who were supplied postoperatively with a knee brace for 4 (range 3-6) weeks. Group B included 39 patients for whom a brace was not used. The median age was 27 (range 16-48) years in group A and 26 (range 14-51) years in group B. The median time period between the injury and the index operation was 24 (range 3 150) months in group A and 18 (range 3-360) months in group B. All 78 patients were re-examined by two independent observers after a median follow-up period of 25 (range 23-28) months in group A and 24 (range 22-27) months in group B. The median KT-1000 total side-to-side difference between the reconstructed and the uninjured knees at 89 N was 3 (range -5.5-11) mm in group A and 3 (range -7-10) mm in group B (NS). When the anterior translation was tested separately at 89 N, the corresponding values were 3 (range -4-13) mm in group A and 3 (range -5-10) mm in group B (NS). The median one-leg hop quotient was 95% (range 50%-167%) of the uninjured leg in group A and 92% (range 64%-119%) in group B (NS). The median Lysholm score was 89 (range 39-100) points in group A and 85 (range 37-100) points in group B (NS). In group A, 27/39 (69%) patients and in group B 21/39 (54%) patients were classified as excellent or good (NS). The median Tegner activity level was 7 (range 3-9) in group A and 6 (range 3-9) in group B (NS). Using the IKDC scale, 27/39 (69%) in group A and 24/39 (62%) in group B were classified as normal or nearly normal (NS). The median sick leave in group A was 62 (range 0-357) days and 59 (range 0-243) days in group B (NS). No serious complications occurred during the first 6 postoperative weeks. Two serious complications were, however, registered after the 6th postoperative week. One patient in group A sustained a rupture of the reconstructed ACL 8 weeks postoperatively (3 weeks after removing the brace), and one patient in group B sustained an undislocated patellar fracture during the 7th postoperative week after a fall. This study indicates that the use of a postoperative rehabilitation brace after an arthroscopic ACL reconstruction did not appear to influence either objective stability or subjective function by the 2-year follow-up. PMID- 9335028 TI - Fatigue after eccentric quadriceps femoris work produces earlier gastrocnemius and delayed quadriceps femoris activation during crossover cutting among normal athletic women. AB - Athletic women are at greater risk of anterior cruciate ligament (ACL) injury than men. Twenty, healthy, athletic women were evaluated for the effect of preferred stance limb isokinetic quadriceps femoris and hamstring fatigue from eccentric work compared with controls on the activation onset of vastus medialis, rectus femoris, vastus lateralis, the medial hamstrings, biceps femoris, and gastrocnemius muscles. Following 3 weeks of crossover cut training, subjects were tested for fatigue effects (5 subjects/week, 3 conditions, 1 condition/day, order effect controlled) on muscle activation onsets prior to crossover cut landing heelstrike (mixed model, ANOVA, P < 0.05). Fatigue from eccentric quadriceps femoris work produced delayed vastus medialis (P = 0.03), rectus femoris (P = 0.007), and vastus lateralis (P = 0.03) activation onsets compared with control, but did not differ compared to hamstring fatigue. Neither hamstring nor quadriceps femoris fatigue produced differences (P > 0.05) in medial hamstring or biceps femoris activation onsets compared to control. Quadriceps femoris fatigue from eccentric work produced earlier gastrocnemius activation onsets (P = 0.048) than control, but did not differ for hamstring fatigue. The gastrocnemius appears to provide synergistic and compensatory dynamic knee stabilization in closed kinetic chain function during quadriceps femoris fatigue. This finding in a normal group at high risk of ACL injury while performing a maneuver with a high ACL injury risk supports gastrocnemius inclusion in knee rehabilitation and conditioning programs and suggests the need for comparative evaluations of knee injured/reconstructed subjects. PMID- 9335029 TI - Holmium: YAG-laser-assisted arthroscopy versus conventional methods for treatment of the knee. Two-year results of a prospective study. AB - The results of 320 arthroscopic procedures are reported here, in which laser surgery using the holmium: YAG laser is compared with conventional mechanical methods. The patients were followed-up during a 2-year period and the data analyzed in a prospective study. The following knee injuries were included: meniscal lesion, chondromalacia, combined meniscal/cartilage lesion, rheumatoid synovialitis and patellofemoral pain syndrome. Because strict inclusion criteria were used, the patient collective is homogenous. Gender, age, injured side, intrasurgical diagnosis, and the initial values of the Lysholm score (modified after Klein) are congruous. After 2 years, the results of the laser collective were significantly improved, whereas the results for the conventional collective, especially for chondromalacia and synovialitis, did not show the same improvement. Analysis of the effect of various instruments and the laser system itself show differing results for the various knee disorders. The hemostatic effect of the holmium: YAG laser was excellent during surgery of all knee disorders, including surface treatment. Operating time for laser surgery was not prolonged, in contrast to what is often claimed. This study shows that chondromalacia, combined meniscal-cartilage lesions, and chronic rheumatoid synovialitis are treated more effectively and with better results with the holmium: YAG laser than with conventional arthroscopic methods. Furthermore, laser treatment of lateral retinacular release can be considered to be better than mechanical techniques. No significant advantage can be found for using the laser during meniscectomy. Lasers are useful for treating smaller, hard-to-reach joints and lower the risk of iatrogenic cartilage damage. The holmium: YAG laser is a suitable instrument for arthroscopic surgery. PMID- 9335030 TI - Lesions of the scapholunate ligaments in acute wrist trauma--arthroscopic diagnosis and minimally invasive treatment. AB - The final result of the treatment of distal intra-articular radius fractures depends both on the accuracy of the fracture reduction and on the presence of additional carpal injuries. In particular, lesions of the intrinsic ligaments usually lead to severe degenerative damage of the wrist joint if they are missed primarily. With the introduction of wrist arthroscopy, these tears can be evaluated and treated earlier. Since 1993 arthroscopically assisted treatment has been performed in 23 patients with distal intra-articular fractures of the radius (mainly C-fractures according to the AO classification system or group VII and VIII fractures according to Frykman). Scapholunate (SL) tears were found in 11 patients (47.8%), 7 of whom showed marked instability intraoperatively and were stabilised at the time of surgery. PMID- 9335031 TI - Optimization of high-frequency electrosurgery of the meniscus. AB - In an in vitro study of the sheep's meniscus, the possibilities to reduce the thermal tissue necrosis caused by high-frequency electrosurgery of the meniscus were studied. Under standardized conditions, machine-made sections through the meniscus were cut using electrodes of various thicknesses and different settings of the electric generator. The thermal tissue necrosis near the cut through the meniscus was determined using light microscopy and image analysis on the specimen stained according to Masson-Goldner. With a constant voltage of 250 V tissue necrosis was significantly less for electrodes of a diameter of 0.5 mm than for 1.5-mm electrodes (187.3 microns as compared with 368.0 microns). For electrosections carried out using a commercially available electrode with a low voltage of 250 V as well as with a power-controlled generator, tissue necrosis was also significantly less than with a constant high voltage of 395 V (181.4 microns and 210.2 microns as compared with 325.0 microns). Thus, an effective reduction of thermal tissue necrosis in arthroscopic partial meniscectomy is possible when thin electrodes and power-controlled generators are used. PMID- 9335032 TI - Effect of basic fibroblast growth factor on the healing of defects in the canine anterior cruciate ligament. AB - The effect of basic fibroblast growth factor (bFGF) on the healing of partial anterior cruciate ligament (ACL) lacerations was investigated in 17 adult mongrel canines. The defect was created in the midsubstance of both ACLs using a biopsy punch. In the bFGF group, a bFGF-impregnated pellet was sutured to the infrapatellar fat pad close to the defect. In the control group, the same pellet without bFGF was used. The healing process was evaluated macroscopically and histologically at 1, 3, 6, and 24 weeks postoperatively. In the bFGF group, a pannus-like tissue which contained abundant blood vessels extended into the defect from the adjacent synovial tissue in the early postoperative phase. Histological examination of the tissue which filled the defect revealed initial remodeling processes with a decreased number of cells and better orientation of the collagen fibers at 6-24 weeks. On the other hand, in the control group, poor healing processes were observed at each examination period. This study demonstrated that the application of a bFGF-impregnated pellet may enhance the healing potential of a partially injured ACL. Enhanced neovascularization and the formation of granulation tissue induced by bFGF early in the healing process accounted for the increased healing response. PMID- 9335033 TI - Pharmacogenetics and drug metabolism. PMID- 9335034 TI - Discoveries in complex biosystems. PMID- 9335035 TI - Thalidomide reconsidered. PMID- 9335037 TI - Wrangle over rights to BRCA1. PMID- 9335036 TI - Amylin's pramlintide best of bad bunch of diabetes drugs. PMID- 9335038 TI - Transgenic factor VIII: the milky way and beyond. PMID- 9335039 TI - An eye for an eye: new models of genetic ocular disease. PMID- 9335040 TI - Say cheese! The maturing science of starter cultures. PMID- 9335041 TI - Interleukin-6: an antagonizing problem becomes a solution. PMID- 9335042 TI - Turning poison eaters inside out. PMID- 9335043 TI - Laying the foundations for personalized medicines. PMID- 9335044 TI - From bench to business: career opportunities in biotechnology. PMID- 9335045 TI - Applications of the green fluorescent protein in cell biology and biotechnology. AB - The recent emergence of an autofluorescent protein, the green fluorescent protein (GFP), has opened the door for the convenient use of intact living cells and organisms as experimental systems in fields ranging from cell biology to biomedicine. We present an overview of some of the major applications of GFP, namely its use in protein tagging and in monitoring gene expression as well as its potential in a variety of biological screens. PMID- 9335047 TI - Transgenic pigs produce functional human factor VIII in milk. AB - Deficiency or abnormality of coagulation factor VIII (FVIII) causes a bleeding disorder called hemophilia A. Treatment involves FVIII concentrates prepared from pooled human plasma or recombinant FVIII (rFVIII) prepared from mammalian cell culture. The cost of highly purified FVIII or rFVIII is a major factor in hemophilia therapy and restricts prophylaxis. We have sought to generate a new source of rFVIII by targeting expression of the human FVIII cDNA to the mammary gland of transgenic pigs using the regulatory sequences of the mouse whey acidic protein gene. The identity of processed heterodimeric rFVIII was confirmed using specific antibodies, by thrombin digestion and activity assays. The secretion of as much as 2.7 micrograms/ml of rFVIII in milk was over tenfold higher than in normal plasma. Up to 0.62 U/ml of rFVIII was detected in an assay in which rFVIII restored normal clotting activity to FVIII-deficient human plasma. PMID- 9335046 TI - Genetically engineered large animal model for studying cone photoreceptor survival and degeneration in retinitis pigmentosa. AB - Patients with retinitis pigmentosa (RP) typically develop night blindness early in life due to loss of rod photoreceptors. The remaining cone photoreceptors are the mainstay of their vision; however, over years or decades, these cones slowly degenerate, leading to blindness. We created transgenic pigs that express a mutated rhodopsin gene (Pro347Leu). Like RP patients with the same mutation, these pigs have early and severe rod loss; initially their cones are relatively spared, but these surviving cones slowly degenerate. By age 20 months, there is only a single layer of morphologically abnormal cones and the cone electroretinogram is markedly reduced. Given the strong similarities in phenotype to that of RP patients, these transgenic pigs will provide a large animal model for study of the protracted phase of cone degeneration found in RP and for preclinical treatment trials. PMID- 9335048 TI - Food-grade controlled lysis of Lactococcus lactis for accelerated cheese ripening. AB - An attractive approach to accelerate cheese ripening is to induce lysis of Lactococcus lactis starter strains for facilitated release of intracellular enzymes involvement in flavor formation. Controlled expression of the lytic genes lytA and lytH, which encode the lysin and the holin proteins of the lactococcal bacteriophage phi US3, respectively, was accomplished by application of a food grade nisin-inducible expression system. Simultaneous production of lysin and holin is essential to obtain efficient lysis and concomitant release of intracellular enzymes as exemplified by complete release of the debittering intracellular aminopeptidase N. Production of holin alone leads to partial lysis of the host cells, whereas production of lysin alone does not cause significant lysis. Model cheese experiments in which the inducible holinlysin overproducing strain was used showed a fourfold increase in release of L-Lactate dehydrogenase activity into the curd relative to the control strain and the holin-overproducing strain, demonstrating the suitability of the system for cheese applications. PMID- 9335049 TI - Design of thermolabile bacteriophage repressor mutants by comparative molecular modeling. AB - Comparative molecular modeling was performed with repressor protein Rro of the temperate Lactococcus lactis bacteriophage r1t using the known 3D-structures of related repressors in order to obtain thermolabile derivatives of Rro. Rro residues presumed to stabilize a nonhomologous but structurally conserved hydrophobic pocket, which was shown to be important for thermostability of the Escherichia coli bacteriophage lambda repressor CI, were randomized. Of the derivatives that exhibited various temperature-sensitive phenotypes, one was shown to hold promise for both fundamental and industrial applications that require the controlled production of (heterologous) proteins in L. lactis. PMID- 9335050 TI - Biodegradation of organophosphorus pesticides by surface-expressed organophosphorus hydrolase. AB - Organophosphorus hydrolase (OPH) was displayed and anchored onto the surface of Escherichia coli using an Lpp-OmpA fusion system. Production of the fusion proteins in membrane fractions was verified by immunoblotting with OmpA antisera. Inclusion of the organophosphorus hydrolase signal sequence was necessary for achieving enzymatic activity on the surface. More than 80% of the OPH activity was located on the cell surface as determined by protease accessibility experiments. Whole cells expressing OPH on the cell surface degraded parathion and paraoxon very effectively without any diffusional limitation, resulting in sevenfold higher rates of parathion degradation compared with whole cells with similar levels of intracellular OPH. Immobilization of these live biocatalysts onto solid supports could provide an attractive means for pesticide detoxification in place of immobilized enzymes, affording a reduced diffusional barrier. PMID- 9335051 TI - Overexpression of glutathione S-transferase/glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress. AB - Transgenic tobacco seedlings that overexpress a cDNA encoding an enzyme with both glutathione S-transferase (GST) and glutathione peroxidase (GPX) activity had GST and GPX-specific activities approximately twofold higher than wild-type seedlings. These GST/GPX overexpressing seedlings grew significantly faster than control seedlings when exposed to chilling or salt stress. During chilling stress, levels of oxidized glutathione (GSSG) were significantly higher in transgenic seedlings than in wild-types. Growth of wild-type seedlings was accelerated by treatment with GSSG, while treatment with reduced glutathione or other sulfhydryl-reducing agents inhibited growth. Therefore, overexpression of GST/GPX can stimulate seedling growth under chilling and salt stress, and this effect could be caused by oxidation of the glutathione pool. PMID- 9335052 TI - Plant resistance to fungal infection induced by nontoxic pokeweed antiviral protein mutants. AB - Pokeweed antiviral protein (PAP), a 29-kD protein isolated from Phytolacca americana inhibits translation by catalytically removing a specific adenine residue from the large rRNA of the 60S subunit of eukaryotic ribosomes. Transgenic plants expressing PAP are resistant to a broad spectrum of plant viruses. Nontoxic PAP mutants have been isolated by random mutagenesis and selection in yeast. One of these mutants, PAP-X, had a point mutation at the active-site (E176V) that abolished enzymatic activity, and another mutant, delta C25PAP, had a nonsense mutation near the C-terminus (W237stop) that deleted 25 C terminal amino acids. Unlike the wild-type PAP, expression of neither mutant was toxic to transgenic plants. We show that both class I (basic) and class II (acidic) isoforms of pathogenesis-related (PR) proteins are overexpressed in transgenic plants expressing PAP and the nontoxic PAP mutants. Although PR proteins are constitutively expressed, no increase in salicylic acid levels was detected. Homozygous progeny of transgenic plants expressing either PAP or the nontoxic PAP mutants displayed resistance to the fungal pathogen Rhizoctonia solani. These results show that expression of PAP or the nontoxic PAP mutants activates multiple plant defense pathways independently of salicylic acid and confers resistance to fungal infection. The C-terminal 25 amino acids of PAP, which are required for toxicity in vivo, are not critical for resistance to viral or fungal infection, indicating that toxicity of PAP can be separated from pathogen resistance. PMID- 9335053 TI - Induction of interleukin-6 (IL-6) autoantibodies through vaccination with an engineered IL-6 receptor antagonist. AB - Neutralization of cytokine activity by monoclonal antibodies or receptor antagonists is beneficial in the treatment of immune and neoplastic diseases, but the necessity for continuous parenteral delivery of these anticytokine agents poses considerable practical limitations. A viable alternative is to induce a neutralizing antibody response. Using transgenic mice with high circulating levels of human interleukin-6 (hIL-6), we show that injection of the hIL-6 receptor antagonist Sant1 (an IL-6 variant with seven amino-acid substitutions) induces a strong anti-hIL-6 antibody response. The elicited antibodies bind circulating hIL-6 with very high affinity, totally masking it, and neutralize hIL 6 bioactivity both in vitro and in vivo. PMID- 9335054 TI - A two-component expression system that responds to inflammatory stimuli in vivo. AB - A therapeutic dilemma often complicates the management of inflammatory diseases; the benefits gained from reducing inflammation must be balanced against the potentially harmful consequences of chronic immunosuppression. Gene therapy might address this dilemma by producing anti-inflammatory proteins in response to a patient's endogenous signals, so that recombinant drug production is linked to the intensity and duration of the inflammatory condition. To test this, we have developed inflammation-inducible systems for regulating recombinant protein production in vivo. We describe a two-component expression construct in which (1) the murine complement factor 3 (C3) promoter regulates production of the human immunodeficiency virus (HIV) transactivator of transcription (Tat), and (2) the Tat protein then stimulates protein expression from genes inserted downstream of the the HIV promoter. When incorporated into a nonreplicating adenovirus (Ad.C3 tat/HIV-luc) and studied in a murine model, the construct produces large amounts of recombinant protein in vivo in response to two different inflammatory stimuli. PMID- 9335055 TI - Human hematopoietic progenitor cell isolation based on galactose-specific cell surface binding. AB - The ability to isolate functional populations of hematopoietic progenitor cells is important to the process of hematopoietic cell transplantation and to the understanding of hematopoietic cell biology in health and disease. We show that a subpopulation of human bone marrow hematopoietic cells bearing the pan hematopoietic antigen CD34 also binds galactose-conjugated proteins. This lectin positive sub-population represents approximately 0.1 to 0.5% of the total bone marrow cells, and contains 100% of the hematopoietic progenitor cells. The galactose-binding lectin on these cells is specific for this sugar. Additionally, highly proliferative hematopoietic progenitor cells with very primitive phenotypes, including a newly identified progenitor cell that produces multiple lineages, express this lectin. PMID- 9335056 TI - Artificial intelligence for drug design. PMID- 9335057 TI - Progressive multifocal leukoencephalopathy in AIDS: initial and follow-up CT and MRI. AB - We sought to determine the value of follow-up CT and MRI in patients with acquired immuno-deficiency syndrome (AIDS) and progressive multifocal leukoencephalopathy (PML). We reviewed 50 CT and 19 MRI examinations performed in 21 biopsy- or autopsy-proven cases of PML; 17 patients had follow-up examinations (mean time 5.9 weeks). The radiological examinations were correlated with pathological findings at autopsy. On initial imaging studies, 73 lesions were found. On follow-up, the most striking feature was rapid progression in both size and number of the lesions (from a mean of 3.2 to 6.9 per patient). One third of the patients showed increasing mass effect. A central area suggesting necrosis, of variable size, was found in 12/16 patients. Autopsy revealed macroscopic necrotic changes in the lesions in 11/16 patients. PMID- 9335058 TI - Idiopathic hypertrophic pachymeningitis: spectrum of the disease. AB - Hypertrophic pachymeningitis is extremely rare. It is a fibrosing inflammatory process which involves the dura mater, including the tentorium. Numerous pathological entities produce thickening of the pachymeninges, so that idiopathic hypertrophic pachymeningitis is a diagnosis of exclusion. We describe four patients with idiopathic hypertrophic pachymeningitis who had varied clinical presentation. Imaging studies revealed diffuse thickening of the pachymeninges; in one patient there was extensive dural sinus thrombosis. Since no identifiable cause was found, the cases were labelled as idiopathic. PMID- 9335059 TI - Crossed cerebellar diaschisis in chronic Broca's aphasia. AB - The cerebellum has anatomical connections to the cerebral cortex, through which it can affect language function. To study these connections, we investigated patients with chronic Broca's aphasia using MRI and single-photon emission computed tomography (SPECT). We selected 15 such patients (9 male, 6 female, aged 17-64 years, mean age 56 years) from 30 chronically aphasic patients. Using the results of SPECT, we divided them into patients with (group 1) and without (group 2) crossed cerebellar diaschisis (CCD). We compared the language function of the two groups. Patients in group 1 showed classical Broca's aphasia, while patients in group 2 showed mainly word-finding difficulty. Patients with CCD hat infarcts involving the lower part of the frontal gyrus but patients without CCD did not, which suggests that this region may have functional and anatomical connections with the cerebellum. Our findings support the notion that the cerebellum contributes to language. PMID- 9335060 TI - Tumour blood flow and partition coefficients: correlation with grade of cerebral gliomas using xenon-enhanced computed tomography. AB - It is possible to underestimate the grade of nonenhancing cerebral tumours on conventional contrast-enhanced MRI or CT. Differentiation of high- and low-grade gliomas by measurement of the brain-blood partition coefficient lambda (T lambda) with Xe-enhanced CT (XeCT) has been reported. We assessed the practical applications of XeCT in suspected low-grade astrocytomas. We examined 15 patients with tumours which showed no contrast enhancement on conventional MRI and CT, using XeCT. Tumour blood flow (TBF) and T lambda were calculated. Fourteen patients underwent surgery, one patient had a biopsy. We recognized three histological groups. While T lambda differed significantly between them, TBF did not. Group 1 contained grade II-III astrocytomas and T lambda was 0.77; group 2 contained grade I-II astrocytomas with T lambda 1.14, and group 3 four oligodendrogliomas in which a T lambda of 1.50 was found. PMID- 9335061 TI - Dynamic MRI of meningiomas and schwannomas: is differential diagnosis possible? AB - We studied 23 patients with meningiomas and 14 with schwannomas using dynamic spin-echo (TR/TE 200/15 ms) MRI. Histologically the meningiomas were classified according to the 1993 WHO classification. Serial images were obtained every 30s for 210s after rapid injection of gadopentetate dimeglumine (0.1 mmol/kg). The contrast-enhancement ratio (CER) was divided into three patterns; a sharp rise with a peak within 60 s (A), a relatively rapid increase with a peak between 60 and 210s (B), a slow increase without a peak (C). The patterns were correlated with the histology of the tumors. The signal intensities of the tumours on T2 weighted images were also analyzed and correlated with the dynamic patterns. Meningiomas had more varied dynamic patterns than schwannomas. Almost half of the meningiomas showed pattern A, and one third pattern C. Of six meningiothelial meningiomas showed pattern A; all schwannomas and fibrous meningiomas showed pattern C. Various patterns were observed in transitional meningiomas. Of the 8 meningiomas showing pattern C, only one gave high signal on T2-weighted images, and could not be differentiated from the schwannomas. Thus, one third of meningiomas could not be differentiated from schwannomas by the dynamic contrast enhancement alone. However, when this was combined with the signal intensity on T2-weighted images, most meningiomas could be differentiated from schwannomas. PMID- 9335062 TI - Malignant evolution of presumed benign lesions in the brain in neurofibromatosis: case report. AB - We report a patient suffering from neurofibromatosis type 1 in whom neoplasms developed from the areas of altered signal which are generally considered benign and typical of the disease. MRI, despite two previous examinations 3 and 2 years before development of the tumour, gave no clue to an unfavourable outcome. PMID- 9335063 TI - Neurofibromatosis type 1: brain stem tumours. AB - We describe the clinical and imaging findings of brain stem tumours in patients with neurofibromatosis type 1 (NF1). The NF1 patients imaged between January 1984 and January 1996 were reviewed and 25 patients were identified with a brain stem tumour. Clinical, radiographical and pathological results were obtained by review of records and images. Brain stem tumour identification occurred much later than the clinical diagnosis of NF1. Medullary enlargement was most frequent (68%), followed by pontine (52%) and midbrain enlargement (44%). Patients were further subdivided into those with diffuse (12 patients) and those with focal (13 patients) tumours. Treatment for hydrocephalus was required in 67% of the first group and only 15% of the second group. Surgery was performed in four patients and revealed fibrillary astrocytomas, one of which progressed to an anaplastic astrocytoma. In 40% of patients both brain stem and optic pathway tumours were present. The biological behaviour of brain stem tumours in NF1 is unknown. Diffuse tumours in the patients with NF1 appear to have a much more favourable prognosis than patients with similar tumours without neurofibromatosis type 1. PMID- 9335064 TI - Case report. MRI and MR angiography of basilar artery dissection in a child. AB - MRI showed a pontine infarct and mural thickening of intermediate signal in T1 weighted images proximal to occlusion of the basilar artery in a 10-year-old boy. Two days later the mural thickening was of high signal, consistent with methaemoglobin formation and MR angiography (MRA) showed nonspecific lack of flow in the mid-segment of the basilar artery, which corresponded to a tapered occlusion at arteriography. MRI is more useful than MRA for noninvasive diagnosis of basilar artery dissection. PMID- 9335065 TI - Transient nonketotic hyperglycinaemia: ultrasound, CT and MRI: case report. AB - We report a case of transient nonketotic hyperglycinaemia in which radiography correlated closely with clinical and biochemical findings. Only 5 patients have been previously described with this transient from of nonketotic hyperglycinaemia. Among the radiographic findings, thinning of the corpus callosum is the most characteristic. PMID- 9335066 TI - Internal derangements of the temporomandibular joint: comparison of assessment with three-dimensional gradient-echo and spin-echo MRI. AB - Our purpose was to assess the diagnostic accuracy of three-dimensional (3D) gradient-echo (GRE) MRI in the diagnosis of internal derangements of the temporomandibular joint (TMJ). We studied 140 joints in 70 patients with TMJ internal derangements. We obtained 3D-GRE and spin-echo (SE) images in the closed mouth position; the images were reviewed for disc displacement as well as bone and cartilage abnormalities. The 3D-GRE and SE images were interpreted independently by different radiologists. The sensitivity and specificity of 3D GRE imaging for assessing mediolateral disc displacement was 100%, whereas the sensitivity and specificity of sagittal SE images were 82% and 72%, respectively (P < 0.001). The 3D-GRE images also demonstrated more bone abnormalities (in 112 condyles or 80%) than did SE images (in 79 condyles or 56%) (P < 0.001). Furthermore, 3D-GRE imaging revealed articular cartilage abnormalities in 46 condyles (33%) that were not visible on SE images. The frequency of pain was significantly higher in joints with bone and cartilage abnormalities (P < 0.05 and P < 0.001, respectively). For assessment of disc displacement, 3D-GRE images were superior to sagittal SE images alone, and comparable to combined sagittal and coronal SE images, while for bone and cartilage abnormalities, they were superior to sagittal and coronal SE images. PMID- 9335068 TI - Undernutrition during suckling changes the sensitivity to haloperidol and chlorpromazine in two behavioural measures in weaning rats. AB - Undernutrition during critical periods of development may cause changes in the behavioural responses of rats to centrally acting drugs. In the present study, the effects of undernutrition during suckling on the behavioural responses of 21 days-old rats to chlorpromazine (0, 2.5, 5, 10 and 20 mg/kg) or haloperidol (0, 0.125, 0.25, 0.5, 1 or 2 mg/kg) were examined. Locomotion was assessed at 1 hr 30 min., 4 hr 30 min., 7 hr 30 min, and 10 hr 30 min., and catalepsy was scored at 3 hr, 6 hr and 9 hr after drug administration. Drug was injected on two consecutive days. On day 1, saline-treated undernourished rats showed significantly greater locomotion activity than did normal rats. The neuroleptic-induced inhibition of locomotor activity in undernourished rats was significantly less than that observed in normal rats from 4 hr 30 min. to 10 hr 30 min. (chlorpromazine) or from 7 hr 30 min. to 10 hr 30 min. (haloperidol). On day 2, a similar trend was observed but only in rats injected with 5 mg/kg chlorpromazine or 0.5, 1, and 2 mg/kg haloperidol. On day 1, the catalepsy scores at 3 hr revealed no significant difference between nutritional groups, but at 6 hr undernourished rats responded significantly less to chlorpromazine or haloperidol. On day 2, undernourished rats were less responsive to neuroleptics than normal rats, but the effect was not so evident as observed on day 1. The present results suggest that the behavioural effects of chlorpromazine and haloperidol are less persistent in undernourished rats, possibly due to differences in drug distribution and elimination, when compared to well-nourished rats. PMID- 9335067 TI - Molecular characteristics of mammalian dopamine receptors. AB - Dopamine receptors belong to a large super-gene family of receptors which are linked to their signal transduction pathways through heterotrimeric G proteins. A variety of signalling events are known to be regulated by dopamine receptors including adenylate cyclase and phospholipase activities and various ion channels. Prior to the advent of molecular cloning technology, dopamine receptors were believed to belong to two subtypes, D1 and D2. This distinction was based on both pharmacological and functional criteria. We now know that at least five different dopamine receptors exist although they can still be described as to belonging within "D1" and "D2" subfamilies. The D1 subfamily consists of two receptors-the D1 and D5, whereas the D2, D3 and D4 receptors comprise the D2 subfamily. The cloning and molecular characteristics of these five receptors are described in this review. PMID- 9335069 TI - On the mechanism(s) of cholecystokinin (CCK): receptor stimulation attenuates morphine dependence in mice. AB - In the present study, effect of cholecystokinin (CCK) agonists and on dependence to morphine in mice has been investigated. The influence of dopaminergic, adrenergic, cholinergic and serotonergic on attenuation of naloxone-induced jumping in morphine-dependent mice by CCK agonists were also considered. Mice were treated subcutaneously with morphine (50, 50 and 75 mg/kg) three times daily (10 a.m. 1 p.m. and 4 p.m.) for 3 days, and a last dose of morphine (50 mg/kg) was administered on the 4th day. Withdrawal syndrome (jumping) was precipitated by naloxone (5 mg/kg) which was administered intraperitoneally 2 hr after the last dose of morphine. To study effects of CCK receptor agonists, 10 injection of morphine (3 administrations each day) for dependence and a dose of 5 mg/kg of naloxone for withdrawal induction were employed. The CCK agonists CCK-8 (0.001 0.1 mg/kg), unsulfated CCK-8 (CCK-8U; 0.001-0.1 mg/kg) and caerulein (0.00001 0.01 mg/kg) were able to prevent withdrawal signs precipitated by naloxone (5 mg/kg). Sulpiride and pimozide increased response induced by CCK-8 agonists. The dopamine antagonists also attenuates jumping by themselves. SCH 23390 did not alter the CCK-8 effect, but decreased the jumping by itself. Phenoxybenzamine, propranolol, methysergide and atropine did not change the caerulein effect significantly. However, single administration of atropine increased and methysergide decreased jumping. It is concluded that CCK mechanism(s) may be involved in morphine dependence, and dopaminergic mechanism(s) may interact with CCK in attenuation of naloxone-induced jumping. PMID- 9335070 TI - B lymphocytes in mercury-exposed workers. AB - We have investigated the number of B lymphocytes in mercury-exposed workers. The study group consisted of 33 workers from a mercury-producing plant, mean age 27 years and a mean exposure period 19 months. At the time of testing and for the three previous months, the exposed persons had urinary mercury levels below the currently accepted limit of 50 micrograms g creatinine. A significant reduction in the number of B lymphocytes was observed in the mercury-exposed individuals. We found no correlation between B lymphocytes changes and urinary mercury concentrations, length of exposure or age of the workers. PMID- 9335071 TI - L-methionine induces stage-dependent changes of differentiation and oxidative activity in sea urchin embryogenesis. AB - This study was to investigate developmental toxicity of some selected low molecular weight antioxidants, by utilising sea urchin embryos and gametes as model system. Sea urchin embryos or sperm were exposed at different developmental stages to L-methionine or some selected low molecular weight antioxidants: a) N acetylcysteine; b) L-carnosine; c) L-homocarnosine, and d) L-anserine. L methionine displayed developmental toxicity at levels > or = 10(-5) M, whereas the other agents tested were mostly active at levels > or = 10(-4) M. When embryos were exposed to 10(-4) M L-methionine or N-acetylcysteine at different developmental stages, the most severe effects were exerted by early exposures (0 to 2 hr after fertilisation), whereas later exposures turned to lesser or no effects. Cytogenetic analysis of L-methionine-exposed embryos showed a significant mitogenic effect and increase of mitotic aberrations. Fertilisation success was decreased by L-methionine (10(-6) M to 10(-3) M) added at the moment of fertilisation, with increasing developmental and cytogenetic abnormalities in the offspring. The formation of reactive oxygen species in embryos and gametes was determined by: a) analysing the DNA oxidative product, 8-hydroxy-2' deoxyguanosine (8-OHdG), and b) luminol-dependent chemiluminescence. The results showed that: 1) 8-OHdG levels were increased during embryogenesis; 2) fertilisation was associated with a double-wave luminol-dependent chemiluminescence emission; 3) luminol-dependent chemiluminescence was maximal in cleavage, declining down to zero in plutei, and 4) an embryotoxic L-methionine or N-acetylcysteine level (10(-4) M) turned to a decrease in reactive oxygen species formation. The data suggest that L-methionine- or N-acetylcysteine-induced developmental toxicity is confined to early stages. A role for oxidative activity is suggested in modulating cell differentiation and embryogenesis, consistent with antioxidant-induced damage to early life stages. PMID- 9335072 TI - The efficacy of monopyridinium (2-PAAM, 2-PAEM) and bispyridinium (obidoxime, HI 6) oximes against mevinphos in mice. AB - The efficacy of two new monopyridinium oximes (2-PAAM, 2-PAEM) and two bispyridinium oximes (obidoxime. HI-6) was tested in combination with atropine sulphate against acute poisoning with the organophosphorus insecticide mevinphos in mice. When mice were treated two min. after mevinphos poisoning, no significant differences in the therapeutic effect of tested oximes were observed. The oximes increased the 24 hr LD50 values of mevinphos about two times in comparison with the 24 hr LD50 values of mevinphos in mice protected with atropine sulphate alone and more than three times in comparison with non-treated intoxicated animals. On the other hand, both monopyridinium oximes were significantly more efficacious than HI-6 and as efficacious as obidoxime when they were administered 30 sec. after mevinphos poisoning. Both monopyridinium oximes and obidoxime increased the 24 hr values of mevinphos almost three times in comparison with the 24 hr values of mevinphos in mice protected with atropine sulphate alone and about twenty-five times in comparison with non-treated intoxicated animals, while the oxime HI-6 less than two times in comparison with the 24 hr values of mevinphos in mice protected with atropine sulphate alone and about fifteen times in comparison with non-treated intoxicated animals. Use of new monopyridinium oximes seems to be the improvement in the antidotal treatment of poisoning with organophosphorous insecticide mevinphos in comparison with HI-6 but not in comparison with obidoxime when oximes are used in equimolar doses. PMID- 9335073 TI - Interaction of chymotrypsin with carbetocin ([1-deamino-1-monocarba-2-O methyltyrosine]-oxytocin). AB - The results of the present study describe the course of reaction and the products following chymotrypsin treatment of the oxytocin analogue carbetocin ([1-deamino 1-monocarba-2-O-methyltyrosine]-oxytocin). The metabolites were analyzed and identified through TLC, HPLC and mass spectrometry. The main product emerging after treatment of carbetocin with chymotrypsin is 9-desglycineamide carbetocin indicating preferential hydrolysis of the peptide bond between leucine at position 8 and carboxyterminal glycineamide. At the same time the stability of the bond between tyrosine at position 2 and isoleucine at position 3 appears significantly enhanced through the alkylation of the hydroxyl group of tyrosine. PMID- 9335074 TI - MK-801 potentiates morphine-induced impairment of memory consolidation in mice: involvement of dopaminergic mechanisms. AB - The purpose of the present research was to study the interaction between the non competitive NMDA receptor antagonist MK-801 and morphine in memory consolidation. The involvement of dopamine (DA) mechanisms in this interaction was also studied. Four sets of experiments were carried out with CD1 mice in a one-trial inhibitory avoidance task with post-training injections of drugs. In a first series of experiments post-training administration of morphine or of the non-competitive NMDA receptor antagonist MK-801 impaired memory consolidation. In the second set of experiments the memory consolidation impairment exerted by MK-801 was potentiated by the administration of the D1 dopamine (DA) receptor antagonist SCH 23390 and by that of the D2 DA receptor antagonist (-)-sulpiride. In the third set of experiments, administration of a dose of MK-801 ineffective by itself potentiated the memory impairment exerted by morphine. In the fourth series of experiments, similar ineffective doses of the D1 DA receptor agonist SKF 38393 or of the D2 DA receptor agonist LY 171555 antagonized the impairment of memory consolidation produced by MK-801 and morphine in combination, suggesting the involvement of dopaminergic mechanisms. PMID- 9335075 TI - Differences in extracellular dopamine concentrations in the nucleus accumbens during response-dependent and response-independent cocaine administration in the rat. AB - Studies indicate that nucleus accumbens (NAcc) dopamine neurotransmission is involved in the reinforcing and direct effects of cocaine. The present study was initiated to explore further the relationship of NAcc extracellular dopamine concentrations ([DA]e) and cocaine self-administration using a yoked littermate design. In the first experiment, one rat from each litter was trained to self administer cocaine i.v. (SA: 0.33 mg/inf) under a fixed ratio 2 schedule, while a second rat received simultaneous infusions of cocaine yoked to the infusions of the SA (YC). NAcc [DA]e and cocaine concentrations ([COC]) were assessed during the test sessions using in vivo microdialysis combined with microbore HPLC procedures. [DA]e and [COC] were significantly elevated in the SA and YC groups during the self-administration session; however, [DA]e were greater in the SA group compared to the YC group in the first hour of the session, even though [COC] were not significantly different. On the following day, the rats previously allowed to self-administer cocaine were administered response-independent cocaine infusions yoked to the infusion pattern from the previous day. [DA]e were significantly elevated above baseline levels during the session but were significantly less than concentrations obtained when cocaine was self administered by these subjects. [COC] during the sessions were not significantly different between the two days. Baseline [DA]e were not significantly different between the SA and YC groups or between Day 1 and Day 2. Furthermore, there was no significant difference in the in vitro probe recovery between one and two days following probe implantation. These results suggest that the context in which cocaine was administered significantly altered the neurochemical response to equivalent brain concentrations of cocaine. NAcc [DA]e was significantly increased when the delivery of cocaine infusions was contingent on the behavior of the rat, indicative of a role in the neural processes underlying cocaine reinforcement. PMID- 9335076 TI - Tolerance and cross-tolerance to morphine-like stimulus effects of mu opioids in rats. AB - The purpose of these experiments was to examine the relationship of agonist relative efficacy to the pattern of tolerance and cross-tolerance to the morphine like stimulus effects of three opioid agonists. Rats were trained to discriminate 3.2 mg/kg morphine from saline under fixed-ratio 15 schedule of food reinforcement. Morphine, nalbuphine, and fentanyl produced dose-dependent increases in morphine-like stimulus effects and decreases in response rates. Repeated treatment with 20 mg/kg per day morphine increased the ED50 for stimulus control by fentanyl, morphine, or nalbuphine two-, four-, or 40-fold, respectively. Repeated treatment with 64 mg/kg per day nalbuphine increased the ED50 for stimulus control for morphine by two-fold, but lower or higher treatment doses had no significant effect. Treatment with 100 mg/kg per day nalbuphine increased the ED50 for nalbuphine by six-fold. Repeated treatment with 0.22 mg/kg per day fentanyl increased the ED50 for stimulus control by fentanyl or morphine by approximately two-fold. Comparisons among treatment conditions suggested that magnitude of tolerance to morphine-like stimulus effects did not vary as an inverse function of the relative efficacy of the agonist used for repeated treatment. Rather repeated morphine and fentanyl treatments produced comparable tolerance, whereas repeated nalbuphine treatment did not evoke substantial tolerance. Comparisons within treatment conditions, however, suggested that magnitude of tolerance may vary inversely with relative efficacy of the agonist tested for morphine-like stimulus effects. During treatment with morphine or fentanyl, greater tolerance was observed to the morphine-like stimulus effects of the lower efficacy agonist relative to the higher efficacy agonist. PMID- 9335077 TI - Behavioural and pharmacological characterisation of the canopy stretched attend posture test as a model of anxiety in mice and rats. AB - The behavioural element, stretched attend posture (SAP), is an important component of the "risk-assessment" repertoire of defensive behaviour in rodents. The present experimental paradigm was devised as a novel and simple method of eliciting high levels of SAP in mice and rats. The SAP test apparatus comprised an elevated black Perspex circular platform. A smaller clear red Perspex circular "Canopy" was supported directly above the platform by a central pillar, thus dividing the platform into an inner, dimly lit covered zone and an outer, brightly lit exposed zone. In both the rat and mouse version of this model, vehicle-treated animals exhibited a marked preference for exploring the covered zone and also exhibited high baseline levels of SAP, particularly at the covered zone boundary whilst they investigated the exposed zone. In the mouse SAP test, the benzodiazepine receptor agonists, diazepam (0.5 mg/kg s.c.) and chlordiazepoxide (2 mg/kg s.c.), and the 5-HT1A receptor agonists, buspirone (1 and 3 mg/kg s.c.), ipsapirone (3 mg/kg s.c.) and 8-OH-DPAT (0.2 mg/kg s.c.), all significantly decreased the frequency of SAP without impairing motor activity. In the rat SAP test, diazepam (0.5 mg/kg s.c.) significantly decreased, whilst the anxiogenic 5-HT2C/1B receptor agonist, mCPP (0.25 and 0.5 mg/kg s.c.), significantly increased, the frequency of SAP. Ipsapirone (3 mg/kg s.c.) induced a non-specific behavioural inhibition. These data suggest that the "Canopy" SAP test is a useful paradigm to investigate risk assessment behaviour in both rats and mice, and may provide a sensitive novel rodent model of anxiety. PMID- 9335078 TI - Effect of fluvoxamine on platelet 5-HT2A receptors as studied by [3H]lysergic acid diethylamide ([3H]LSD) binding in healthy volunteers. AB - Alterations in platelet 5-HT2A receptor characteristics have been reported in major depression as well as in other psychiatric diseases, and some effort has been made to utilize platelet 5-HT2A receptor status as a biological correlate to antidepressant drug response. In order to investigate whether treatment with a selective serotonin reuptake inhibitor affects platelet 5-HT2A receptors, we have studied platelet [3H]lysergic acid diethylamide ([3H]LSD) binding in healthy subjects treated with fluvoxamine in increasing dosage once weekly for 4 weeks. After 1 week of fluvoxamine treatment (25 mg/day), both Bmax and Kd were significantly lower than before the start of the treatment (19.9 versus 25.5 fmol/mg protein, P = 0.005 for Bmax; 0.45 versus 0.93 nM, P = 0.006 for Kd). Bmax returned to baseline during week 2, whereas Kd was lower than the baseline value throughout the treatment period. After discontinuation of fluvoxamine treatment, there was a significant increase in Kd (0.50 nM before discontinuation vs. 1.14 nM after discontinuation; P = 0.001), but not in Bmax. The study demonstrates that fluvoxamine affects platelet 5-HT2A receptor status irrespective of underlying psychiatric disease, and that this effect is evident already after 1 week at a subtherapeutic fluvoxamine dose. PMID- 9335079 TI - Dizocilpine-like discriminative stimulus effects of competitive NMDA receptor antagonists in mice. AB - Several non-competitive NMDA receptor ion channel blockers, competitive NMDA antagonists and compounds acting at other sites on the NMDA receptor complex were examined for their ability to substitute for the discriminative stimulus effects of dizocilpine. Swiss-Webster mice were trained with food to discriminate the non competitive NMDA receptor antagonist, dizocilpine (0.17 mg/kg), from saline in a T-maze. Mice rapidly acquired the discrimination with minimal amounts of drugs required for training and testing. Several non-competitive antagonists dose dependently substituted for dizocilpine with a rank order of potency of dizocilpine > TCP > (-)-MK-801 > SKF 10,047 > dextrorphan > PCP. There was a positive correlation between the potencies of the compounds that substituted for dizocilpine and their previously reported affinities for the [3H]dizocilpine binding site of the NMDA receptor ion channel. Compounds acting at other sites on the NMDA receptor complex, including NMDA, the partial agonist at the strychnine insensitive glycine site, ACPC, and the polyamine antagonist, ifenprodil, failed to substitute fully. In addition, the AMPA antagonist, NBQX, the monoamine uptake inhibitor, cocaine, and the GABAA receptor agonists, diazepam and phenobarbital, failed to substitute fully for dizocilpine. However, like the ion channel blockers, the competitive NMDA antagonists, CGS 19755, NPC 17742, (+/-)CPP and LY 233536 dose-dependently substituted for dizocilpine. The competitive antagonist, LY 274614, and its active enantiomer, LY 235959, failed to substitute for dizocilpine, each producing severe disruptions in locomotor activity. That most of the competitive antagonists substituted for dizocilpine is in accordance with other behavioral data (e.g., ataxia, locomotor activity) documenting similarities in the effects of non-competitive and competitive antagonists. These findings are also consistent with results of clinical investigations suggesting overlap in the behavioral and subjective profiles of competitive and non-competitive NMDA blockers. PMID- 9335080 TI - 5-HT1A receptors in lithium-induced conditioned taste aversion. AB - Experiments were carried out using rats to investigate whether 5-HT1A neural mechanisms are involved in lithium-induced conditioned taste aversion (CTA). We found that the 5-HT1A antagonists p-MPPI and pindolol caused CTA similar to that produced by LiCl. The 5-HT1A agonist 8-OH-DPAT counteracted lithium-induced CTA. Pindolol dose-dependently abolished effects of 8-OH-DPAT on LiCl-induced CTA. These findings support the notion that lithium has antagonistic actions on 5-HT1A receptors. Inhibition of 5-HT synthesis by PCPA failed, however, to prevent lithium-induced CTA. Evidently, mechanisms other than those governed solely by 5 HT are also involved in lithium-induced CTA. PMID- 9335081 TI - Influence of clonidine on psychopathological, endocrine and respiratory effects of cholecystokinin tetrapeptide in patients with panic disorder. AB - The influence of clonidine pretreatment on psychopathological, endocrine and respiratory effects of cholecystokinin tetrapeptide (CCK-4) was characterized. Patients with panic disorder (DSM-III-R) were given 50 micrograms CCK-4 i.v. at 1100 hours on 2 separate study days. In a randomized double-blind design they were additionally infused with 150 micrograms clonidine or placebo from 1040 to 1110 hours. After CCK-4 all patients experienced symptom attacks. No effects of clonidine on panic psychopathology or blood gas parameters were observed. After CCK-4, in the clonidine condition the pituitary release of adrenocorticotropin (ACTH) and prolactin was seemingly enhanced compared to placebo. Our results suggest that CCK-4-induced panic attacks are not suppressible by presynaptic alpha-2 receptor stimulation. Moreover, they point to a synergistic postsynaptic action of clonidine to CCK-4 upon pituitary hormone secretion. The diverging sites of action might possibly explain the discrepancies of psychopathological alterations and stress hormone secretion. PMID- 9335082 TI - Kappa opioid mediated locomotor activity in the preweanling rat: role of pre- and postsynaptic dopamine receptors. AB - Treatment with a non-selective DA receptor agonist (i.e., NPA) has previously been shown to attenuate the kappa opioid mediated locomotor activity of preweanling rats. The purpose of the present study was to determine whether stimulation of D1-like or D2-like receptors is responsible for this behavioral effect and whether the critical DA receptors are located pre- or postsynaptically. To assess these questions, 17-day-old rats were injected with saline, the D2/D3 agonist quinpirole (0.1, 0.3, or 1.0 mg/kg, i.p.), or the D1 agonist SKF 38393 (7.5, 15, or 30 mg/kg, i.p.), 20 min after receiving the kappa opioid agonist U-50,488 (5 mg/kg, s.c.) or saline. Results showed that the locomotor activating effects of U-50,488 were blocked by the D2/D3, but not the D1, receptor agonist. To dissociate the effects of DA autoreceptors and postsynaptic receptors, 17-day-old rats were given alpha-methyl-DL-p-tyrosine (AMPT reduces endogenous DA stores) prior to U-50,488 or amphetamine (1.5 mg/kg, s.c.) treatment. Interestingly, AMPT (which reduced DA levels by more than 80%) fully attenuated amphetamine-induced locomotor activity, while having little effect on U-50,488-induced locomotion. In addition, quinpirole blocked the locomotor activating effects of U-50,488 in rats acutely depleted of DA. When considered together, these results indicate that kappa opioid stimulation enhances locomotor activity regardless of presynaptic DA levels. Similarly, quinpirole appears to attenuate U-50,488-induced locomotor activity by stimulating postsynaptic D2-like receptors, since the D2/D3 agonist inhibited kappa opioid mediated behavior independent of endogenous DA levels. PMID- 9335083 TI - Ethanol, like psychostimulants and morphine, causes long-lasting hyperreactivity of dopamine and acetylcholine neurons of rat nucleus accumbens: possible role in behavioural sensitization. AB - Repeated treatment of rats with ethanol (1 g/kg, once daily for 15 days) enhanced the locomotor effect of morphine, 3 weeks post-treatment. This ethanol-induced long-term behavioural sensitization to morphine was associated with an increase in the electrically evoked release of [3H]dopamine (DA) and [14C]acetylcholine (ACh) from nucleus accumbens slices. A similar enhanced responsiveness of accumbal dopaminergic and cholinergic neurons to depolarization was apparent 3 weeks after repeated morphine, amphetamine or cocaine administration. Prior ethanol exposure also caused a long-term enhancement of electrically evoked release of [3H]DA and [14C]ACh from slices of the caudate-putamen. Unlike the locomotor effect of morphine, that of amphetamine was not enhanced in ethanol pretreated rats. These data indicate that ethanol administration may cause long term behavioural sensitization associated with adaptive changes in dopaminergic and cholinergic neurons of rat nucleus accumbens and caudate-putamen. Furthermore, an enhanced reactivity of nucleus accumbens dopaminergic nerve terminals and dopamine-sensitive cholinergic neurons appears to be a common long term neuroadaptive effect of distinct types of addictive drugs. However, since repeated ethanol exposure did not cause a long-term increase in the locomotor effect of amphetamine, these neuroadaptations may not always be sufficient to cause long-lasting behavioural (cross-)sensitization. PMID- 9335084 TI - Evidence that the enhancement of dopamine function by repeated electroconvulsive shock requires concomitant activation of D1-like and D2-like dopamine receptors. AB - In this study, the behavioural response to dopamine D1-like receptor agonists (SKF 38393, SKF 81297 and SKF 77434) and D2-like receptor agonists (quinpirole and RU 24213), administered alone and in combination to rats treated repeatedly with electroconvulsive shock (five ECS over 10 days) or sham, was tested. Agonist induced behaviour was monitored by automated activity meters and direct observation using a checklist scoring method. Repeated ECS (compared to sham controls) had no significant effect on the behavioural response to SKF 38393 (7.5 mg/kg s.c.), SKF 81297 (0.2 mg/kg s.c.), SKF 77434 (0.1 mg/kg s.c.), quinpirole (0.1 and 0.25 mg/kg s.c.) or RU 24213 (0.3 mg/kg s.c.), when administered alone. In contrast, repeated ECS markedly increased locomotion (activity counts and scores) induced by the non-selective dopamine agonist apomorphine (0.5 mg/kg SC) and by co-administration of a D1-like agonist plus a D2-like agonist [SKF 38393 (7.5 mg/kg s.c.) plus quinpirole (0.25 mg/kg s.c.), SKF 81297 (0.2 mg/kg s.c.) plus quinpirole (0.1 mg/kg s.c), and SKF 77434 (0.1 mg/kg s.c.) plus RU 24213 (0.3 mg/kg s.c.)]. This ECS-induced enhancement of dopamine-mediated behaviour was observed for up to 3 weeks after cessation of ECS treatment. In addition, ECS also enhanced the locomotor response to intra-accumbens SKF 38393 plus quinpirole (0.4 and 1.0 microgram/side, respectively). These results provide evidence that the enhancement of dopamine function by repeated ECS requires concomitant stimulation of both D1-like and D2-like receptors, and that this effect is long lasting. PMID- 9335085 TI - Enhancement of dopamine-mediated behaviour by the NMDA antagonists MK-801 and CPP: similarities with repeated electroconvulsive shock. AB - The behavioural effect of dopamine D1-like receptor agonists (SKF 38393, SKF 81297) and a D2-like receptor agonist (quinpirole), administered alone and in combination, was tested in rats pretreated with a single injection of an NMDA antagonist (MK-801, CPP) or vehicle. Agonist-induced behaviour was monitored by automated activity meters and direct observation using a checklist scoring method. Pretreatment with MK-801 (0.05 mg/kg, SC, 30 min) had no significant effect (compared to controls) on the behavioural response to SKF 38393 (7.5 mg/kg SC), SKF 81297 (0.2 mg/kg SC) or quinpirole (0.1 and 0.25 mg/kg SC) administered alone. In contrast, MK-801 markedly increased locomotion (activity counts and scores) induced by co-administration of a D1-like plus a D2-like agonist [SKF 38393 (7.5 mg/kg) plus quinpirole (0.25 mg/kg), SKF 81297 (0.2 mg/kg) plus quinpirole (0.1 mg/kg)]. The behavioural response to the non-selective dopamine agonist apomorphine (0.5 mg/kg SC) was also enhanced by MK-801. Pretreatment with CPP (0.1 mg/kg SC, 30 min) also significantly increased the locomotor response to co-administration of SKF 38393 plus quinpirole administered alone, but had no effect on the behavioural response to separate injection of these agonists. MK 801 (0.05 mg/kg SC, 30 min) also enhanced the behavioural response to bilateral injection into the nucleus accumbens of SKF 38393 plus quinpirole (1.0 plus 0.4 microgram/side, respectively). These data suggest that in the intact rat, the enhancement of dopamine-mediated behaviour by either MK-801 or CPP requires concomitant stimulation of D1-like and D2-like receptors, possible located within the nucleus accumbens. The effect of these NMDA antagonists on dopamine function is similar to that of repeated electroconvulsive shock (ECS), indicating that one of the actions of ECS may be to reduce NMDA receptor function. PMID- 9335086 TI - Relationship between the CYP2D6 genotype and the steady-state plasma concentrations of trazodone and its active metabolite m-chlorophenylpiperazine. AB - The relationship between the cytochrome P450 (CYP) 2D6 genotype and the steady state plasma concentrations (Css) of trazodone and its active metabolite m chlorophenylpiperazine (mCPP) was studied in 54 depressed Japanese patients receiving trazodone 150 mg at bedtime. By use of allele-specific PCR analysis, the wild type allele, three mutated alleles causing absent enzyme activity (CYP2D6A, CYP2D6B and CYP2D6D) and one mutated allele causing decreased enzyme activity (CYPZD6 Ch) were identified. The means (ranges) of the Css of trazodone, corrected to the median body weight in 17 cases with no mutated allele, 27 cases with one mutated allele and 10 cases with two mutated alleles, were 556 (281 1115), 643 (302-1362) and 671 (234-1418) ng/ml, respectively, while the values of mCPP were 60 (35-121), 65 (33-99) and 58 (38-112) ng/ml, respectively. Neither the Css of trazodone (F = 0.80, P = 0.45) nor that of mCPP (F = 0.49, P = 0.61) significantly differed among the three groups. The present study thus suggests that the CYP2D6 genotype cannot predict the Css of these compounds. PMID- 9335087 TI - Frontal 5-HT2A receptors studied in depressive patients during chronic treatment by selective serotonin reuptake inhibitors. AB - To investigate adaptative changes of 5-HT2A receptors induced by SSRIs, six patients chronically treated for a depressive episode (four with fluoxetine, two with fluvoxamine) were studied with PET and [18F]setoperone. They were compared to eight untreated depressive patients. The mean frontal to cerebellum radioactivity concentration ratio, an index of the [18F]setoperone specific binding to 5-HT2A receptors, was higher in treated than in untreated patients, when age was taken into account. This suggests that chronic treatment by SSRIs could induce an up-regulation of the 5-HT2A receptors, and that 5-HT2A receptor down-regulation is not a common mechanism for the therapeutic effects of all serotoninergic antidepressive drugs. PMID- 9335088 TI - Previous treatment as a confounding variable in studies with novel antipsychotics: two cases of high dopamine-2 receptor occupancy with quetiapine. AB - Previous treatment can be a confounding variable in studies with novel antipsychotics. Quetiapine is a new antipsychotic substance with a low affinity for dopamine-2 (D2) receptors. Preliminary SPECT and PET investigations revealed only a low striatal D2 receptor occupancy rate. However, we present two cases of high striatal D2 receptor occupancy (51% and 71%) measured with 123I IBZM SPECT during quetiapine monotherapy. Both patients had previously received continuous treatment with typical neuroleptics. We present evidence that the previous antipsychotic therapy influenced D2 receptor binding of 123I IBZM during quetiapine treatment weeks after cessation of typical neuroleptics. This might be of importance for the design of clinical trials and brain imaging studies in the future. PMID- 9335089 TI - Computed tomography of hereditary multifocal renal cystadenocarcinomas in German shepherd dogs. AB - The purpose of the study was to characterize the renal lesions of hereditary renal cystadenocarcinomas in the German shepherd dog using computed tomography (CT). Fourteen dogs with renal cystadenocarcinomas and nodular dermatofibrosis, and two unaffected dogs were studied. There were nine dogs with spontaneous disease and seven dogs from a test mating. The characteristic renal CT findings of renal cystadenocarcinomas were bilateral multiple cysts and tumor masses of various sizes. The earliest changes were detected between 4 and 5 years of age and the smallest cysts measured 2-3 mm in diameter. Abdominal CT examination gives a wide field view and excellent anatomic images of the kidneys. It is easy to differentiate between cysts and solid tumors. CT examination is useful for the early detection of renal cystadenocarcinomas and for screening suspected carrier dogs before breeding. PMID- 9335090 TI - Evaluation of the variably ossified collateral cartilages of the distal phalanx and adjacent anatomic structures in the Finnhorse with computed tomography and magnetic resonance imaging. AB - Six Finnhorse cadaver forefeet were selected to represent radiographically different types and grades of ossification of the collateral cartilages of the distal phalanx. These cartilages and adjacent tissues were evaluated with computed tomography (CT) and high field magnetic resonance imaging (MRI). In CT the internal structure of the cartilages was consistent, but in MRI some differences were noted. The shape of the collateral cartilages and their ligamentous attachments varied. The border between ossified and non-ossified cartilage appeared distinct, with considerable variation in the extent of the ossified area in regard to the cross-sectional area of the cartilage. Ossification originating from the palmar processes and extending in the proximal/palmaroproximal direction, without separate centers of ossification, generally appeared smooth and inactive. Palmar ossification followed the irregular shape of the cartilage. Separate centers of ossification had a medullary cavity or were sclerotic. Presence of a medullary cavity or sclerosis were also found at the base of the cartilages. The incomplete fusion lines between separate centres of ossification and the ossified base of the cartilage varied from congruent and inactive to reactive with marked sclerosis, flared margins and parachondral changes. Incomplete fusion may be clinically significant. Local conformational adaptations of the hoof were also documented with extensive ossification of the collateral cartilage. PMID- 9335091 TI - Radiographic study of distal radial physeal closure in thoroughbred horses. AB - Monthly radiography was performed to study distal radial physeal closure in ten male and ten female Thoroughbred horses. The height, thoracic circumference and metacarpus circumference were also measured. Distal radial physeal closure time was sooner in females than males, and took 701 +/- 37 and 748 +/- 55 days respectively. PMID- 9335092 TI - The effect of patient positioning on mural filling defects during double contrast cystography. AB - Different radiographic findings may be observed during double contrast cystography due to patient positioning affecting the distribution of positive and negative contrast media. A mass lesion was created in the urinary bladder of a canine cadaver to allow evaluation of the effect of patient positioning on the appearance of a mass during double contrast cystography. The mass appeared as a filling defect only on those views where positive contrast medium surrounded the mass. Otherwise, the mass appeared as a summation. Additionally, a patient is described illustrating the effect of patient positioning on detecting mural filling defects during double contrast cystography. PMID- 9335093 TI - Radiographic diagnosis: canine ureteral calculus. PMID- 9335094 TI - Computed tomography and magnetic resonance imaging of cavernous sinus enlargement in a dog with unilateral exophthalmos. AB - Computed tomography (CT), magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) were performed on a dog with a two year history of unilateral exophthalmos occurring two years following head trauma. On CT images, an expansile enhancing mass was present along the right intracranial cavernous sinus and extended through the orbital fissure into the retrobulbar space. With MRI, the structure appeared as a signal void due to the presence of rapidly flowing blood. Gadolinium enhancement of the adjacent brain was not present. A vascular origin of the lesion was confirmed with MRA. Based on the CT and MRI findings, the enlarged cavernous sinus and associated ophthalmic plexus were believed to represent an arterialized aneurysm, most likely the result of traumatic arteriovenous fistulization. Treatment consisted of surgical enucleation. At the time of this report, 29 months later, the dog remains free of clinical signs. PMID- 9335095 TI - Magnetic resonance imaging appearance of intracranial hemorrhage secondary to cerebral vascular malformation in a dog. AB - A 14-year-old dog developed an acute onset of depression, disorientation, left hemiparesis,left hemianopia, left facial hypoesthesia, and a tendency to turn to the right. Based on these findings, a lesion affecting the right forebrain was suspected. Magnetic resonance imaging showed a mass within the right cerebral hemisphere resulting in compression of the right lateral ventricle and shifting the longitudinal fissure to the left. The lesion was hyperintense on T1-weighted images and hyperintense with focal regions of hypointensity on proton density-, and T2-weighted images, consistent with a subacute hemorrhage. At necropsy, there was a hematoma in the parietal portion of the right cerebral hemisphere. The hemorrhage was surrounded by numerous thin-walled veins, most likely a venous malformation. Magnetic resonance imaging of intracranial hemorrhage is reviewed. PMID- 9335096 TI - Dilated coronary sinus in a dog with persistent left cranial vena cava. AB - This paper describes the electrocardiographic, echocardiographic (two dimensional, M-mode, contrast and Doppler) and non-selective angiocardiographic features in a 3 year old female Beagle with dilated coronary sinus due to persistent left cranial vena cava. Negative P waves in leads III and aVR and a positive P wave in lead aVL were seen. Echocardiographically, a hypoechoic circular structure was seen between the left atrium and the pericardium in the area where the coronary sinus is located. A velocity pattern with two peaks was obtained, one systolic with velocity = 0.44 +/- 0.05 m/sec and the other diastolic with velocity = 0.27 +/- 0.01 m/sec. By M-mode echocardiography, at level of the aorta and the left atrium, a linear structure was identified between the left atrium and the pericardium; this structure was characterized by phasic movements of the anterior wall during the cardiac cycle. Following a left cephalic vein injection of saline, bubbles were seen within the coronary sinus; when saline was injected into the right cephalic vein, bubbles were also seen within the coronary sinus and right atrium and ventricle. Non-selective angiocardiography confirmed a dilated coronary sinus with persistent left cranial vena cava. The right cranial vena cava was absent. The dog was clinically normal and the unusual vessel was an incidental finding. PMID- 9335097 TI - Partial avulsion of the origin of the cranial cruciate ligament in a 4 year-old dog. AB - A four year-old intact male Dalmatian was referred to the veterinary teaching hospital at Louisiana State University for acute, non-weight-bearing left hindlimb lameness of three weeks duration. Information supplied by the referring veterinarian indicated the lameness was first diagnosed and treated seven months previously, but recurred three weeks ago. External rotation of the left stifle, mild discomfort upon stifle flexion, mild to moderate muscle atrophy and palpable joint effusion were noted during physical examination. Slight cranial drawer movement with a soft end-point was discovered during manipulation of the left stifle. A triangular bone fragment and thickened, confluent intracapsular soft tissues were observed on radiographs of the stifle. Radiographically, moderate degenerative changes suggested chronicity. This report describes the clinical and radiographic findings of a rarely reported partial avulsion of the origin of the cranial cruciate ligament in a skeletally mature dog. PMID- 9335098 TI - Upper airway obstruction in a llama caused by aberrant nasopharyngeal bots (Cephenemyia sp.). AB - A 9 month old female llama was presented with inspiratory dyspnea. Radiographically, there was a large soft tissue mass nearly occluding the nasopharynx. During endoscopic examination three nasopharyngeal bots were identified embedded in the mass. The larvae were removed and the patient treated with ivermectin. The patient was discharged one week later free of clinical respiratory disease. In follow-up radiographs made 6 weeks later, only residual radiopacity in the area of the mass remained. PMID- 9335099 TI - Side lobes and grating lobes artifacts in ultrasound imaging. AB - Side lobes and grating lobes are both unwanted parts of the ultrasound beam emitted off axis that produce image artifacts due to error in positioning the returning echo. The purpose of this study was to reproduce artifacts associated with side lobes and grating lobes in vitro using different transducer types and recognize these artifacts in vivo. A phantom, composed of a water bath, a metallic wire, and a wooden tongue depressor, was imaged using a linear array, a curved linear a vector array, and a sector mechanical transducer. When imaging the metallic wire in a transverse plane, an echogenic artifact was constantly seen on each side of the wire, with a shape and intensity variable with the transducer type. The artifact was curvilinear and concave (linear and curved linear arrays), or curvilinear and convex (vector array and the mechanical transducer). When the tongue depressor was imaged in a longitudinal plane, the artifact was a straight line (linear array), a curved convex line (curved array), a series of convex curvilinear echo (vector array) or a small convex curvilinear echo (mechanical transducer). In vivo situations similar to the phantom experiment were investigated using clinical patients. Artifacts produced in vitro were recognized in vivo when a highly reflective object (urinary bladder wall) was imaged adjacent to an anechoic region (urine). These artifacts corresponded to the principle of secondary ultrasound lobes, and were therefore interpreted as such. PMID- 9335100 TI - Ultrasonographic localization of a caudal vena cava thrombus in a dog with leishmaniasis. AB - A dog with visceral leishmaniasis developed rear limb edema, and distension of the caudal epigastric veins. Glomerular disease with nephrotic syndrome and hypercoagulable state was diagnosed. Sonographically there was massive thrombosis of the caudal vena cava. PMID- 9335101 TI - Ultrasonographic appearance of orthotopic ureterocele in a dog. AB - A three-year-old, intact male, Siberian husky was evaluated for a two day history of dysuria. Sonographically there was an anechoic cyst-like structure in the urinary bladder. The abnormality appeared to be a 'cyst within a cyst', which is a characteristic ultrasonographic feature of ureterocele in humans. Ultrasonography may be a useful means of establishing a diagnosis of ureterocele in dogs. PMID- 9335111 TI - The quality of self-concept in old age. AB - Self-concept in old age was investigated. The goal of the work was to reveal the contents of self-concept, that is, what is included in the sphere of self assessment, and also to analyze the traditional parameters of self-assessment: its height, stability, adequacy. Specific features of self-concept in old age were found. They concern the contents of self-concept and are characterized by retrospective direction of self-analysis. Total lowering of the level of self assessment and modification of its structure including real, ideal, attainable positions were found. As compared with previous periods of the life-cycle, self concept becomes less stable and adequate. Besides the general peculiarities, old people's self-concept is characterized by wide individual variations and sex differences. PMID- 9335112 TI - Assignments, secondary structure and dynamics of the inhibitor-free catalytic fragment of human fibroblast collagenase. AB - Fibroblast collagenase (MMP-1), a 169-residue protein with a molecular mass of 18.7 kDa, is a matrix metalloproteinase which has been associated with pathologies such as arthritis and cancer. The assignments of the 1H, 15N, 13CO and 13C resonances, determination of the secondary structure and analysis of 15N relaxation data of the inhibitor-free catalytic fragment of recombinant human fibroblast collagenase (MMP-1) are presented. It is shown that MMP-1 is composed of a beta-sheet consisting of five beta-strands in a mixed parallel and antiparallel arrangement (residues 13-19, 48-53, 59-65, 82-85 and 94-99) and three alpha-helices (residues 27-43, 112-124 and 150-160). This is nearly identical to the secondary structure determined from the refined X-ray crystal structures of inhibited MMP-1. The major difference observed between the NMR solution structure of inhibitor-free MMP-1 and the X-ray structures of inhibited MMP-1 is the dynamics of the active site. The 2D 15N-1H HSQC spectra, the lack of information in the 15N-edited NOESY spectra, and the generalized order parameters (S2) determined from 15N T1, T2 and NOE data suggest a slow conformational exchange for residues comprising the active site (helix B, zinc ligated histidines and the nearby loop region) and a high mobility for residues Pro138 Gly144 in the vicinity of the active site for inhibitor-free collagenase. In contrast to the X-ray structures, only the slow conformational exchange is lost in the presence of an inhibitor. PMID- 9335113 TI - Alterations in chemical shifts and exchange broadening upon peptide boronic acid inhibitor binding to alpha-lytic protease. AB - alpha-Lytic protease, a bacterial serine protease of 198 amino acids (19 800 Da), has been used as a model system for studies of catalytic mechanism, structure function relationships, and more recently for studies of pro region-assisted protein folding. We have assigned the backbones of the enzyme alone, and of its complex with the tetrahedral transition state mimic N-tert-butyloxycarbonyl-Ala Pro-boro Val, using double- and triple-resonance 3D NMR spectroscopy on uniformly 15N- and 13C/15N-labeled protein. Changes in backbone chemical shifts between the uncomplexed and inhibited form of the protein are correlated with distance from the inhibitor, the displacement of backbone nitrogens, and change in hydrogen bond strength upon inhibitor binding (derived from previously solved crystal structures). A comparison of the solution secondary structure of the uninhibited enzyme with that of the X-ray structure reveals no significant differences. Significant line broadening, indicating intermediate chemical exchange, was observed in many of the active site amides (including three broadened to invisibility), and in a majority of cases the broadening was reversed upon addition of the inhibitor. Implications and possible mechanisms of this line broadening are discussed. PMID- 9335114 TI - Solution conformation and dynamics of a tetrasaccharide related to the Lewis(x) antigen deduced by NMR relaxation measurements. AB - 1H-NMR cross-relaxation rates and nonselective longitudinal relaxation times have been obtained at two magnetic fields (7.0 and 11.8 T) and at a variety of temperatures for the branched tetrasaccharide methyl 3-O-alpha-N-acetyl galactosaminyl-beta-galactopyranosyl-(1-->4)[3-O-alph a -fucosyl]-glucopyranoside (1), an inhibitor of astrocyte growth. In addition, 13C-NMR relaxation data have also been recorded at both fields. The 1H-NMR relaxation data have been interpreted using different motional models to obtain proton-proton correlation times. The results indicate that the GalNAc and Fuc rings display more extensive local motion than the two inner Glc and Gal moieties, since those present significantly shorter local correlation times. The 13C-NMR relaxation parameters have been interpreted in terms of the Lipari-Szabo model-free approach. Thus, order parameters and internal motion correlation times have been deduced. As obtained for the 1H-NMR relaxation data, the two outer residues possess smaller order parameters than the two inner rings. Internal correlation times are in the order of 100 ps. The hydroxymethyl groups have also different behaviour, with the exocyclic carbon on the glucopyranoside unit showing the highest S2. Molecular dynamics simulations using a solvated system have also been performed and internal motion correlation functions have been deduced from these calculations. Order parameters and interproton distances have been compared to those inferred from the NMR measurements. The obtained results are in fair agreement with the experimental data. PMID- 9335115 TI - Performance of a neural-network-based determination of amino acid class and secondary structure from 1H-15N NMR data. AB - A neural network which can determine both amino acid class and secondary structure using NMR data from 15N-labeled proteins is described. We have included nitrogen chemical shifts, 3JHNH alpha coupling constants, alpha-proton chemical shifts, and side-chain proton chemical shifts as input to a three-layer feed forward network. The network was trained with 456 spin systems from several proteins containing various types of secondary structure, and tested on human ubiquitin, which has no sequence homology with any of the proteins in the training set. A very limited set of data, representative of those from a TOCSY HSQC and HNHA experiment, was used. Nevertheless, in 60% of the spin systems the correct amino acid class was among the top two choices given by the network, while in 96% of the spin systems the secondary structure was correctly identified. The performance of this network clearly shows the potential of the neural network algorithm in the automation of NMR spectral analysis. PMID- 9335116 TI - Backbone dynamics of oxidized and reduced D. vulgaris flavodoxin in solution. AB - Recombinant Desulfovibrio vulgaris flavodoxin was produced in Escherichia coli. A complete backbone NMR assignment for the two-electron reduced protein revealed significant changes of chemical shift values compared to the oxidized protein, in particular for the flavine mononucleotide (FMN)-binding site. A comparison of homo- and heteronuclear NOESY spectra for the two redox states led to the assumption that reduction is not accompanied by significant changes of the global fold of the protein. The backbone dynamics of both the oxidized and reduced forms of D. vulgaris flavodoxin were investigated using two-dimensional 15N-1H correlation NMR spectroscopy. T1, T2 and NOE data are obtained for 95% of the backbone amide groups in both redox states. These values were analysed in terms of the 'model-free' approach introduced by Lipari and Szabo [(1982) J. Am. Chem. Soc., 104, 4546-4559, 4559-4570]. A comparison of the two redox states indicates that in the reduced species significantly more flexibility occurs in the two loop regions enclosing FMN. Also, a higher amplitude of local motion could be found for the N(3)H group of FMN bound to the reduced protein compared to the oxidized state. PMID- 9335117 TI - 1H, 15N and 13C NMR resonance assignment, secondary structure and global fold of the FMN-binding domain of human cytochrome P450 reductase. AB - The FMN-binding domain of human NADPH-cytochrome P450 reductase, corresponding to exons 3-7, has been expressed at high level in an active form and labelled with 13C and 15N. Most of the backbone and aliphatic side-chain 1H, 15N and 13C resonances have been assigned using heteronuclear double- and triple-resonance methods, together with a semiautomatic assignment strategy. The secondary structure as estimated from the chemical shift index and NOE connectivities consists of six alpha-helices and five beta-strands. The global fold was deduced from the long-range NOEs unambiguously assigned in a 4D 13C-resolved HMQC-NOESY HMQC spectrum. The fold is of the alternating alpha/beta type, with the five beta strands arranged into a parallel beta-sheet. The secondary structure and global fold are very similar to those of the bacterial flavodoxins, but the FMN-binding domain has an extra short helix in place of a loop, and an extra helix at the N terminus (leading to the membrane anchor domain in the intact P450 reductase). The experimental constraints were combined with homology modelling to obtain a structure of the FMN-binding domain satisfying the observed NOE constraints. Chemical shift comparisons showed that the effects of FMN binding and of FMN reduction are largely localised at the binding site. PMID- 9335119 TI - In situ analysis of protein chromatography and column efficiency using magnetic resonance imaging. AB - Magnetic resonance imaging has been used to visualize size-based protein separations inside operating chromatography columns. The effects of flow nonuniformity have been observed and analyzed quantitatively through concentration profiles of tracers measured inside the column. Analysis of these profiles provides local and averaged intracolumn plate height values for characterization of dispersion and flow nonuniformity. The magnetic resonance measurements compare favorably with conventional chromatographic measurements of column efficiency and provide more detailed insights into nonideal column performance. PMID- 9335120 TI - Characterization of liquid chromatographic stationary phases by Raman spectroscopy. Effect of ligand type. AB - This study represents the first Raman spectroscopic characterization of conventional chemically-bonded liquid chromatographic (LC) stationary phases under typical flow-rate and pressure conditions. Raman spectra were obtained for amino propyl (NH2), cyano propyl (CN), phenyl (Ph), octadecyl (C18), octyl (C8), and methyl (C1) chemically-bonded silica-based stationary phases in 100% aqueous mobile phases. The present experimental set-up has allowed Raman spectra of various stationary phase ligands, present in sub-monolayer coverages on the siliceous supports, to be obtained. This study: (1) demonstrates that conventional Raman spectroscopic techniques can be used to study LC stationary phases; (2) presents the experimental set-up, conditions, and approaches utilized to obtain Raman spectra of conventional stationary phases; (3) examines the spectroscopic differences observed for a variety of different types of bonded ligands that are typically used in reversed-phase (RPLC) and normal-phase (NPLC) liquid chromatographic separations; and (4) considers other future studies that are possible with this experimental approach, including mobile phase composition and temperature studies. PMID- 9335118 TI - Assignment and secondary structure of calcium-bound human S100B. AB - The NMR assignments of backbone 1H, 13C, and 15N resonances for calcium-bound human S100B were completed via heteronuclear multidimensional NMR spectroscopic techniques. NOE correlations, amide exchange, 3JHNH alpha coupling constants, and CSI analysis were used to identify the secondary structure for Ca-S100B. The protein is comprised of four helices (helix I, Glu2-Arg20; helix II, Glu31-Asn38; helix III, Gln50-Thr59; helix IV, Phe70-Phe87), three loops (loop I, Glu21-His25; loop II, Glu39-Glu49; loop III, Leu60-Gly66), and two beta-strands (strand I, Lys26-Lys28; strand II, Glu67-Asp69) which form a short antiparallel beta-sheet. Helix IV is extended by approximately one turn when compared to the secondary structures of apo-rat [Drohat et al. (1996) Biochemistry, 35, 11577-11588] and bovine S100B [Kilby et al. (1996) Structure, 4, 1041-1052]. In addition, several residues outside the calcium-binding loops in S100B undergo significant backbone chemical shift changes upon binding calcium which are not observed in the related protein calbindin D9k. Together these observations support previous site-directed mutagenesis, absorption spectroscopy, and cysteine chemical reactivity experiments, suggesting that the C-terminus in Ca-S100B is important for interactions with other proteins. PMID- 9335121 TI - Ultrasensitive near-infrared laser-induced fluorescence detection in capillary electrophoresis using a diode laser and avalanche photodiode. AB - A sensitive fluorescence detector for capillary electrophoresis consisting of a semiconductor near-infrared diode laser and a single photon avalanche diode (SPAD) is described. The sensitivity of this system was demonstrated by the separation and analysis of four tricarbocyanine dyes using capillary electrophoresis and a running buffer consisting of 98% methanol and 2% water with 40 mM borate (pH 9.4). The LOD for the dye, IR-132, was found to be 4.41 zmol with the dynamic range found to be approximately four orders of magnitude in concentration. Based on the sampling volume of the system, the number of molecules actually detected at this LOD was approximately 27. To further demonstrate the utility of this diode-based detector, various amino acids were derivatized with a highly anionic near-IR labelling dye. The conjugates were separated in a running buffer comprised of predominately methanol and a cationic surfactant added to reverse the electroosmotic flow. The LOD values for various amino acids were found to be in the low zmol range. PMID- 9335122 TI - Analysis of gangliosides by capillary zone electrophoresis and capillary zone electrophoresis-electrospray mass spectrometry. AB - Gangliosides, sialic acid(s)-containing glycosphingolipids, were separated by capillary zone electrophoresis and detected with either UV or electrospray mass spectrometry. Several electrolyte system were evaluated for the separation of underivatized gangliosides. The best result was obtained by using 50 mM borate and 50 mM phosphate buffer containing 20 mM alpha-cyclodextrin at pH 9.9. The four major ganglioside forms (GM1, GD1a, GD1b, GT1b) were successfully separated, and, moreover, each ganglioside yielded two peaks, splitting by the difference in chain length of the ceramide moiety. The resolution obtained in CE-UV could not be reproduced in CE-MS because of the incompatibility of the borate/phosphate buffer to ESI-MS. With the use of more volatile buffers, such as ammonium acetate or 2-[N-cyclohexylamino]-ethanesulfonic acid, baseline resolution was obtained for gangliosides having different number of sugars, but the two disialoganglioside isomers, GD1a and GD1b, were coeluted. PMID- 9335123 TI - Bile salt micellar electrokinetic chromatography of bilirubin and related compounds. AB - The interaction of bilirubin, biliverdin, bilirubin dimethyl ester, biliverdin dimethyl ester, xanthobilirubic acid, and xanthobilirubin methyl ester with trihydroxy and dihydroxy bile salt solutions is investigated by micellar electrokinetic chromatography (MEKC). The capacity factor of each compound is measured in solutions of the different bile salts over the pH range of 6.5-9.0. The capacity factor of bilirubin increases with pH below 7 in all bile salt solutions. Biliverdin and xanthobilirubin show essentially identical capacity factors for all bile salts. Biliverdin dimethyl ester and xanthobilirubin methyl ester also have very similar capacity factors, which are greater than those of the carboxy analogs, in trihydroxy bile salts. The capacity factors of these esters are higher in the dihydroxy bile salts, with the capacity factor of biliverdin dimethyl ester being twice that of xanthobilirubin methyl ester. Factors involved in the MEKC analysis of these compounds are discussed. PMID- 9335124 TI - Separation of cardiac glycosides by micellar electrokinetic chromatography and microemulsion electrokinetic chromatography. AB - The interest of micellar electrokinetic chromatography (MEKC) and microemulsion electrokinetic chromatography (MEEKC) for the resolution of four cardiac glycosides is demonstrated. First, the influence of some parameters on the resolution of the solutes in MEKC such as the concentration of the surfactant, pH, addition of organic modifiers and urea is discussed. Then, results are compared with those obtained in MEEKC using different microemulsion compositions. Results indicate that MEEKC possesses several advantages over MEKC for the separation of relatively hydrophobic compounds such as digitalic compounds. First, microemulsions allow a better manipulation of the migration time window and of the retention of the solutes. Moreover, efficiency is improved with shorter analysis time. PMID- 9335125 TI - Use of metal complexation in non-aqueous capillary electrophoresis systems for the separation and improved detection of tetracyclines. AB - Metal complexation in non-aqueous capillary electrophoresis systems was evaluated for the separation and improved detection of tetracycline antibiotics using laser induced fluorescence detection. It was found that three factors were important for the choice of complexing agent: (i) it should be soluble in the organic solvent used for the separation, (ii) it should have a sufficient fast complexing rate so as not to invalidate the electrophoretic separation and, (iii) it should give a large increase in the fluorescence intensity. Mg2+ ions were found to be the most suitable ions for the separation of the tetracyclines as the acetate salt of magnesium is very soluble in organic solvents and only a relatively low current was generated during electrophoresis making it possible to use high concentrations of the complexing metal ion. Metal complexation strongly intensified the fluorescence of tetracyclines and all organic solvents investigated further intensified the fluorescence, e.g. dimethylformamide improved the fluorescence of the oxytetracycline metal complex by a factor of 34 compared to water. However, magnesium acetate was not sufficiently soluble in dimethylformamide and therefore N-methylformamide, improving the fluorescence intensity by only a factor of 9, was used. It was demonstrated that the method can be used for the detection of tetracyclines at the ppb level in milk and plasma. PMID- 9335126 TI - Preparation and liquid chromatographic analysis of propanediol fatty acid esters. AB - Procedures were developed for the synthesis, purification and analysis of propanediol (PD) esters of n-fatty acids (FA). Mono- (MAPD) and di-acylated (DAPD) species were synthesized from PD and FA using an immobilized lipase (Candida antarctica B) in tert-butanol. MAPD and DAPD were isolated using silica gel column chromatography as approximately 95% pure preparations. Normal phase gradient LC provided for resolution of MAPD, DAPD and FA. UV220 nm detection provided a detection limit of about 1 microgram, and a linear response range of up to 2000 micrograms. Response factors were determined for MAPD, DAPD and FA components comprised of n-fatty acyl lengths of 4-16. PMID- 9335127 TI - Comparison of extraction methods and detection systems in the gas chromatographic analysis of volatile carbonyl compounds. AB - High-resolution gas chromatography (HRGC) with electron-capture detection (ECD), nitrogen-phosphorus detection (NPD), flame ionization detection (FID) or with mass spectrometry-selected ion monitoring (MS-SIM) was used in the analysis of volatile carbonyl compounds. Eighteen carbonyl compounds that are typically produced during lipid peroxidation were derivatized quantitatively with pentafluorophenylhydrazine (PFPH) at room temperature, to afford their corresponding water-insoluble hydrazones. These derivatives were extracted into non-polar phases by means of either liquid-liquid extraction (LLE) (hexane) or solid-phase extraction (SPE) on 3 ml C18 octadecyl-bonded phase cartridges. Detection limits of 10(-14) and 10(-12) mol/ml per aldehyde were achieved with the ECD and MS-SIM systems, respectively. The effects of extraction conditions on sensitivity and recovery were determined by performing parallel HRGC-ECD and HRGC MS-SIM analyses of pentafluorophenylhydrazones of the eighteen compounds under study. Recoveries of 51.4-78.9 +/- 1.2-4.5 and 80.9-98.3 +/- 1.0-3.5% were obtained with LLE and SPE, respectively. The method was applied to the analysis of the volatile carbonyl compounds in various heated vegetable oils (corn, palm or sunflower) and to the analysis of volatile aldehydes in human urine. PMID- 9335128 TI - Micelles as pseudo-stationary phases in micellar electrokinetic chromatography. AB - This review article describes some general comments on micellar electrokinetic chromatography (MEKC) from the viewpoint of pseudo-stationary phases and presents a compiled list of surfactants used for MEKC, prepared from published papers. We tried to give comments on some typical surfactants from the practical point of view. PMID- 9335129 TI - Micellar electrokinetic chromatography-mass spectrometry. AB - The combination of micellar electrokinetic chromatography (MEKC) with mass spectrometry (MS) is very attractive for the direct identification of analyte molecules, for the possibility of selectivity enhancement, and for the structure confirmation and analysis in a MS-MS mode. The direct coupling of MEKC with MS can be hazardous due to the effect of nonvolatile MEKC surfactants on MS performance, including the loss of analyte sensitivity and ion source contamination. The possibility of off-line coupling between MEKC and matrix assisted laser desorption/ionization (MALDI)-MS remains to be investigated. Various approaches for on-line coupling MEKC with electrospray ionization (ESI) MS, including the use of high-molecular-mass surfactant, an electrospray-chemical ionization (ES-CI) interface, a voltage switching and buffer renewal system, partial-filling micellar plug and anodically migrating micelles, are reviewed and evaluated. The use of an ES-CI interface is most promising for routine operation of on-line MEKC-MS under the influence of nonvolatile salts and surfactants. The use of a high-molecular-mass surfactant allows the formation of a micellar phase at very low surfactant concentrations and avoids the generation of a high level of background ions in the low m/z region. Alternatively, the application of a partial-filling micellar plug and anodically migrating micelles eliminate the introduction of MEKC micelles into the ESI-MS system. It is possible to directly transfer the conventional MEKC separations to partial-filling MEKC-ESI-MS and MEKC-ESI-MS using anodically migrating micelles without any instrument modifications. PMID- 9335130 TI - Pharmaceutical applications of micelles in chromatography and electrophoresis. AB - This review surveys the use of micelles as separation media in chromatography and electrophoresis. Applications to pharmaceuticals whose molecular masses are relatively small are focused on in this review. In high-performance liquid chromatography (HPLC), chromatography using micelles and reversed-phase stationary phases such as octadecylsilylized silica gel (ODS) columns is known as micellar liquid chromatography (MLC). The main application of MLC to pharmaceutical analysis is the same as in ion-pair chromatography using alkylsulfonate or tetraalkylammonium. In most cases, selectivity is much improved compared with other short alkyl chain ion-pairing agents such as pentanesulfonate or octanesulfonate. Direct plasma/serum injection can be successful in MLC. Separation of small ions is also successful by using gel filtration columns and micellar solutions. In electrophoresis, especially capillary electrophoresis (CE), micelles are used as pseudo-stationary phases in capillary zone electrophoresis (CZE). This mode is called micellar electrokinetic chromatography (MEKC). Most of the drug analysis can be performed by using the MEKC mode because of its wide applicability. Enantiomer separation, separation of amino acids and closely related peptides, separation of very complex mixtures, determination of drugs in biological samples etc. as well as separation of electrically neutral drugs can be successfully achieved by MEKC. Microemulsion electrokinetic chromatography (MEEKC), in which surfactants are also used in forming the microemulsion, is successful for the separation of electrically neutral drugs as in MEKC. This review mainly describes the typical applications of MLC and MEKC for the analysis of pharmaceuticals. PMID- 9335131 TI - Application of micellar electrokinetic capillary chromatography to the analysis of illicit drug seizures. AB - The application of micellar electrokinetic capillary chromatography (MECC) to the analysis of illicit drug seizures is presented. Areas investigated include general screening and qualitative and, in some instances, quantitative analysis of various drugs, including heroin, opium, cocaine, amphetamines, LSD and anabolic steroids. Due to its high efficiency, high selectivity and general applicability, MECC is well suited for forensic drug analyses. PMID- 9335132 TI - Quantitative models of animal learning and cognition. AB - This article reviews the prerequisites for quantitative models of animal learning and cognition, describes the types of models, provides a rationale for the development of such quantitative models, describes criteria for their evaluation, and makes recommendations for the next generation of quantitative models. A modular approach to the development of models is described in which a procedure is considered as a generator of stimuli and a model is considered as a generator of responses. The goal is to develop models that, in combination with many different procedures, produce sequences of times of occurrence of events (stimuli and responses) that are indistinguishable from those produced by the animal under many experimental procedures and data analysis techniques. PMID- 9335133 TI - Pigeon perception and discrimination of rapidly changing texture stimuli. AB - The perception and discrimination of rapidly changing texture stimuli by pigeons was examined in a target localization task. Five experienced pigeons were rewarded for finding and pecking at a randomly placed odd target block of small repeated elements embedded in a larger rectangular array of contrasting distractor elements. On dynamic color test trials, the color of the target, distractor, or both of these regions changed at rates of 100, 250, 500, or 1000 ms per frame. The number of colors appearing within such trials also varied. Pigeons performed well above chance in all test conditions, with target associated changes producing the best discrimination. The results suggest: (a) global relational information can exclusively guide target localization behavior, (b) pigeons can perceptually group and segregate colored textured differences quite rapidly (< or = 100 ms), and (c) pigeons may possess automatic search control processes that can be captured by stimulus-driven changes in the display. PMID- 9335134 TI - Modulation by the stimulus properties of excitation. AB - Three experiments with pigeons investigated the role of excitation in a Pavlovian modulatory paradigm where the reinforcement contingencies of a conditioned stimulus (CS) were signaled by modulatory stimuli. In Experiment 1, excitatory training of the modulator that signaled reinforcement, the positive modulator, had a greater facilitative impact on discrimination learning than did excitatory training of both modulators. Although this could have resulted from simple excitatory summation, Experiment 2 revealed that excitatory training of the negative modulator also enhanced learning more than did excitatory training of both modulators. In Experiment 3, responding to CSs that had come under the control of differentially excitatory modulators was similarly controlled by new stimuli that had received simple differential excitatory training. Results suggest that excitation can play a modulatory role in Pavlovian conditioning. PMID- 9335135 TI - Memory of auditory lists by rhesus monkeys (Macaca mulatta). AB - Monkey auditory memory was tested with increasing list lengths of 4, 6, 8, and 10 sounds. Five-hundred and twenty environmental sounds of 3-s duration were used. In Experiment 1, the monkeys initiated each list by touching the center speaker. They touched 1 of 2 side speakers to indicate whether a single test sound (presented from both side speakers simultaneously) was or was not in the list. The serial-position functions showed prominent primacy effects (good first-item memory) and recency effects (good last-item memory). Experiment 2 repeated the procedure without the list-initiation response and with a variable intertrial interval. The results of both experiments were similar and are discussed in relation to theories and hypotheses of serial-position effects. PMID- 9335136 TI - Effects of number of items on the pigeon's discrimination of same from different visual displays. AB - The pigeon's discrimination of visual displays comprising from 2 to 16 computer icons that were either the same as or different from one another was studied. Discrimination of Same from Different displays improved when the displays contained more icons, both after training with just 16-icon displays (Experiment 1) and after training with 2-, 4-, 8-, 12-, and 16-icon displays (Experiment 2). That improvement was specific to displays of different icons; accuracy to displays of same icons did not differ as a function of icon number. These results were well described by the degree of variability or entropy in multielement visual displays. PMID- 9335137 TI - Scalar timing in temporal generalization in humans with longer stimulus durations. AB - Three experiments investigated temporal generalization performance in humans by using stimulus durations similar to those previously used with rats. In most conditions, chronometric counting was prevented by concurrent shadowing of temporally irregular numbers. Experiment 1 examined performance with visual stimuli, when the standard was 4.0 s long and nonstandard stimuli were spaced either linearly or logarithmically around the standard. Generalization gradients were asymmetrical with linear spacing but symmetrical with logarithmic spacing, a result obtained previously with humans. Experiment 2 used auditory stimuli and varied the standard across values of 2.0, 4.0, 6.0, and 8.0 s. All gradients were asymmetrical, and good superposition was obtained, indicating conformity to scalar timing. Experiment 3 prevented or encouraged chronometric counting by changing instructions, and temporal generalization gradients differed when counting was and was not used. PMID- 9335139 TI - The evolution of small gene clusters: evidence for an independent origin of the maltase gene cluster in Drosophila virilis and Drosophila melanogaster. AB - We analyzed a 5,770-bp genomic region of Drosophila virilis that contains a cluster of two maltase genes showing sequence similarity with genes in a cluster of three maltase genes previously identified in Drosophila melanogaster. The D. virilis maltase genes are designated Mav1 and Mav2. In addition to being different in gene number, the cluster of genes in D. virilis differs dramatically in intron-exon structure from the maltase genes in D. melanogaster, the transcriptional orientation of the genes in the cluster also differs between the species. Our findings support a model in which the maltase gene cluster in D. virilis and D. melanogaster evolved independently. Furthermore, while in D. melanogaster the maltase gene cluster lies only 10 kb distant from the larval cuticle gene cluster, the maltase and larval cuticle gene clusters in D. virilis are located very far apart and on a different chromosome than that expected from the known chromosome arm homologies between D. virilis and D. melanogaster. A region of the genome containing the maltase and larval cuticle gene clusters appears to have been relocated between nonhomologous chromosomes. PMID- 9335138 TI - Drug discrimination under a concurrent fixed-interval fixed-interval schedule. AB - Pigeons were trained to discriminate 5.0 mg/kg pentobarbital from saline under a concurrent fixed-interval (FI) FI schedule of food presentation on which, after pentobarbital administration, responses on one key were reinforced with food under an FI 60-s component and responses on the other key were reinforced under an FI 240-s component. After saline administration, the schedule contingencies on the two keys were reversed. After both pentobarbital and saline, pigeons responded more frequently on the key on which responses had been programmed to produce the reinforcer under the FI 60 component of the concurrent schedule. The schedule was changed to concurrent FI 150 FI 150 s for drug-substitution tests. In each bird, increasing doses of pentobarbital, ethanol, and chlordiazepoxide produced increases in the proportion of responses on the key on which responses had been reinforced under the FI 60 component after pentobarbital administration during training sessions. The proportion of responses on that key was slightly lower for ethanol than for chlordiazepoxide and pentobarbital. At a dose of pentobarbital higher than the training dose, responding decreased on the key that had been reinforced under the FI 60 component during training sessions. Phencyclidine produced less responding on the key programmed under the FI 60-s component than did pentobarbital. Methamphetamine produced responding primarily on the key on which responses had been reinforced under the FI 60-s component after saline administration. PMID- 9335140 TI - Evolution of alcohol dehydrogenase genes in peonies (Paeonia): phylogenetic relationships of putative nonhybrid species. AB - Alcohol dehydrogenase genes were amplified by PCR, cloned, and sequenced from 11 putative nonhybrid species of the angiosperm genus Paeonia. Sequences of five exons and six intron regions of the Adh gene were used to reconstruct the phylogeny of these species. Two paralogous genes, Adh1A, and Adh2, were found; an additional gene, Adh1B, is also present in section Moutan. Phylogenetic analyses of exon sequences of the Adh genes of Paeonia and a variety of other angiosperms imply that duplication of Adh1 and Adh2 occurred prior to the divergence of Paeonia species and was followed by a duplication resulting in Adh1A and Adh1B. Concerted evolution appears to be absent between these paralogous loci. Phylogenetic analysis of only the Paeonia Adh exon sequences, positioning the root of the tree between the paralogous genes Adh1 and Adh2, suggests that the first evolutionary split within the genus occurred between the shrubby section Moutan and the other two herbaceous sections Oneapia and Paeonia. Restriction of Adh1B genes to section Moutan may have resulted from deletion of Adh1B from the common ancestor of sections Oneapia and Paeonia. A relative-rate test was designed to compare rates of molecular change among lineages based on the divergence of paralogous genes, and the results indicate a slower rate of evolution within the shrubby section Moutan than in section Oneapia. This may be responsible for the relatively long branch length of section Oneapia and the short branch length between section Moutan and the other two sections found on the Adh, ITS (nrDNA), and matK (cpDNA) phylogenies of the genus. Adh1 and Adh2 intron sequences cannot be aligned, and we therefore carried out separate analyses of Adh1A and Adh2 genes using exon and intron sequences together. The Templeton test suggested that there is not significant incongruence among Adh1A, ITS, and matK data sets, but that these three data sets conflict significantly with Adh2 sequence data. A combined analysis of Adh1A, ITS, and matK sequences produced a tree that is better resolved than that of any individual gene, and congruent with morphology and the results of artificial hybridization. It is therefore considered to be the current best estimate of the species phylogeny. Paraphyly of section Paeonia in the Adh2 gene tree may be caused by longer coalescence times and random sorting of ancestral alleles. PMID- 9335142 TI - Microsatellite variation and evolution in the Mimulus guttatus species complex with contrasting mating systems. AB - Mutational variability at microsatellite loci is shaped by both population history and the mating system. In turn, alternate mating systems in flowering plants can resolve aspects of microsatellite loci evolution. Five species of yellow monkeyflowers (Mimulus sect. Simiolis) differing for historical rates of inbreeding were surveyed for variation at six microsatellite loci. High levels of diversity at these loci were found in both outcrossing and selfing taxa. In line with allozyme studies, inbreeders showed more partitioning of diversity among populations, and diversity in selfing taxa was lower than expected from reductions in effective population size due to selfing alone, suggesting the presence of either population bottlenecks or background selection in selfers. Evaluation of the stepwise mutation model (a model of DNA replication slippage) suggests that these loci evolve in a stepwise fashion. Inferred coalescent times of microsatellite alleles indicate that past bottlenecks of population size or colonization events were important in reducing diversity in the inbreeding taxon. PMID- 9335141 TI - Relationships among msx gene structure and function in zebrafish and other vertebrates. AB - The zebrafish genome contains at least five msx homeobox genes, msxA, msxB, msxC, msxD, and the newly isolated msxE. Although these genes share structural features common to all Msx genes, phylogenetic analyses of protein sequences indicate that the msx genes from zebrafish are not orthologous to the Msx1 and Msx2 genes of mammals, birds, and amphibians. The zebrafish msxB and msxC are more closely related to each other and to the mouse Msx3. Similarly, although the combinatorial expression of the zebrafish msx genes in the embryonic dorsal neuroectoderm, visceral arches, fins, and sensory organs suggests functional similarities with the Msx genes of other vertebrates, differences in the expression patterns preclude precise assignment of orthological relationships. Distinct duplication events may have given rise to the msx genes of modern fish and other vertebrate lineages whereas many aspects of msx gene functions during embryonic development have been preserved. PMID- 9335143 TI - Sequence convergence in the peptide-binding region of primate and rodent MHC class Ib molecules. AB - In addition to the universally expressed and highly polymorphic class Ia genes, the major histocompatibility complex (MHC) of placental mammals includes class Ib genes that are characterized by restricted expression and low levels of sequence polymorphism. The functional importance of class Ib genes as well as their actual function has long been controversial. Phylogenetic analyses have suggested that there are no orthologous relationships among class Ib loci of mammals belonging to different orders, suggesting that these loci have evolved independently since the placental mammals diverged. Here, we present evidence of convergent evolution at the molecular sequence level in the putative peptide-binding regions (PBRs) of human and mouse class Ib genes. So far, there are few if any convincing examples of convergent evolution at the amino acid sequence level, and such evolution is believed to be likely to occur only as a result of strong positive selection. Because the present case involves the functionally important PBR and because the primate and rodent molecules are known to bind similar peptides, this study represents both a convincing case of molecular-level convergence and evidence that MHC class Ib molecules, although not orthologous, may evolve similar functions convergently. PMID- 9335144 TI - Nucleotide compositional constraints on genomes generate alanine-, glycine-, and proline-rich structures in transcription factors. AB - Correlation between amino acid composition and nucleotide composition is examined. Class III POU transcription factors having higher third GC contents showed higher contents of alanine, glycine, and proline residues encoded by GC rich nucleotides, and vice versa. This correlation was observed even among various types of transcription factors from vertebrates and invertebrates regardless of functional and structural constraints inherent to each protein. Furthermore, reptile class III POU sequences revealed no evolutionary directionality increasing the GC contents from cold- to warm-blooded vertebrates. PMID- 9335146 TI - A comparative study of duplications in bacteria and eukaryotes: the importance of telomeres. AB - The genomes of three bacteria (Haemophilus influenzae, Mycoplasma genitalium, and Escherichia coli) and two eukaryotes (Saccharomyces cerevisiae and Caenorhabditis elegans) were compared. The distribution of their putative open reading frames (ORFs) was studied, and several conclusions were drawn: (1) All of these genomes, even the smallest, exhibit a significant proportion (7%-30%) of duplicated ORFs. This proportion is a function of genome size and appears unrelated to the bacteria/eukaryote division. (2) Some of these ORFs constitute families of up 20 or more members. (3) The levels of sequence similarity within these families are highly variable and their distribution is different among bacteria and eukaryotes. (4) In yeast, there are topological relationships between members of the same family. The paired ORFs are frequently in the same orientation with regard to their respective telomeres and located at comparable distances from them. PMID- 9335145 TI - Low-molecular-weight heat shock proteins in a desert fish (Poeciliopsis lucida): homologs of human Hsp27 and Xenopus Hsp30. AB - The heat shock response of a fish which inhabits a highly stressful environment (Poeciliopsis lucida, a minnow from river systems of the Sonoran desert in northwestern Mexico) was investigated. Cells derived from this fish exhibited a typical heat shock response when exposed to elevated temperature, synthesizing high levels of 90 kDa, 70 kDa, and 30 kDa heat shock proteins (Hsp90, Hsp70, and Hsp30), as well as lower amounts of other heat shock proteins. Additional small heat shock proteins (sHSPs), including Hsp27, were induced after a prolonged heat shock at a time when synthesis of Hsp70 and Hsp30 was decreasing. Characterization of cDNA clones for hsp27 and hsp30 revealed that both are members of the alpha-crystallin/sHSP superfamily but belong to separate lineages within this gene family. The multiple isoforms of P. lucida Hsp30 appear to be members of a multigene family and are most closely related to salmon and Xenopus Hsp30s. In contrast, Hsp27 is highly similar to mammalian and avian sHSPs; it was synthesized as three isoforms which represented differentially phosphorylated forms of a single polypeptide. In Poeciliopsis, the various sHSPs may each perform a subset of the roles attributed to mammalian sHSPs. The conservation of phosphorylation sites in Hsp27 may indicate an involvement in signal transduction to the actin cytoskeleton. The hsp30 genes appear to have diverged more rapidly than the corresponding hsp27 genes; the various members of the Hsp30 family may function as molecular chaperones and, in this role, may be less evolutionarily constrained. Finally, the presence of these two classes of sHSP in a single taxon indicates that these two lineages arose by gene duplication early in the evolution of vertebrates and raises questions about the fate of homologs of Hsp30 in mammals and of Hsp27 in Xenopus. PMID- 9335147 TI - Ancient DNA from amber fossil bees? PMID- 9335148 TI - Two cases of cutaneous phaeohyphomycosis by Alternaria alternata and Alternaria tenuissima. AB - Two cases of cutaneous phaeohyphomycosis, one with a nodular appearance and the other with an erythematous infiltrating patch, are reported in immunocompromised patients. Diagnosis was based on histological examination, which revealed hyphae and round-shaped fungal cells in a granulomatous dermal infiltrate, and on identification of the moulds when biopsy fragments were cultured on Sabouraud dextrose agar without cycloheximide. The pathogens were Alternaria tenuissima in the first case and A. alternata in the second. The fungi were examined by scanning electron microscopy. The patients were checked for bone and lung involvement and were then treated with surgical excision and itraconazole, and itraconazole only, respectively, with clinical and mycological resolution. PMID- 9335149 TI - Onychoprotothecosis due to Prototheca wickerhamii. PMID- 9335150 TI - Appearance of colonies of Prototheca on CHROMagar Candida medium. AB - The microorganisms capable of producing opportunist infections include the yeast like organisms of the genus Candida, and the unicellular algae of the genus Prototheca, which share common features and can, therefore, lead to confusion. Their colonies are almost identical and they grow in the same culture media used routinely in mycology. CHROMagar Candida is a new chromogenic differential isolation medium that facilitates the presumptive differentiation of some of the most clinically important yeast-like organisms. To our knowledge, the use of CHROMagar Candida with Prototheca spp. has not been reported in the literature. This report describes the growth of 151 strains of Prototheca on CHROMagar Candida compared to the growth of a total of 326 well-characterized yeast organisms of the genera Candida, Cryptococcus, Trichosporon, Geotrichum, and Saccharomyces. It is clinically relevant to note that algae of the genus Prototheca (P. wickerhamii, P. zopfii, and P. stagnora) and of the genus Candida parapsilosis produced similar cream-colored colonies on CHROMagar Candida medium. Based on their growth on CHROMagar, a new species of Candida is described, C. zeylanoides, which has blue-green colonies. The colonies of two species of Trichosporon are also differentiated: the blue-green colonies of T. beigelii and the pink colonies of T. capitatum. PMID- 9335151 TI - Paracoccidioidoma: case record and review. AB - A case of solitary pulmonary paracoccidioidal lesion-paracoccidioidoma-is related. It is the first reported case in Brazil. The literature on spontaneously regressive lesion of paracoccidioidomycosis is commented upon. PMID- 9335152 TI - Phenotype and genotype of Candida albicans strains isolated from pregnant women with recurrent vaginitis. AB - Fourteen out of 80 pregnant women receiving prenatal care presented signs and symptoms of recurrent vaginal candidiasis. Candida albicans strains were isolated from 12 patients (85.7%), and these were submitted to morphotyping (morphological characteristics of the colony), antifungal typing (pattern of sensitivity to amphotericin B, 5-fluorcytosine, myconazole, ketoconazole and fluconazole) and genotyping (electrophoretic migration of DNA fragments digested with EcoRI and HinfI). Alteration of morphotype and antifungal type was observed in 50% of the patients, but the genotype of the strains isolated from the same patients at different times was identical in all subjects. The predominant morphotypes presented continuous fringes and the basic changes observed among antifungal types was the emergence of strains resistant to myconazole, which was the drug used for the treatment of the first episode of vaginitis. We conclude that recurrent vaginal candidasis is caused by the persistence of a single yeast genotype that undergoes morphological and behavioral changes in the presence of antifungal agents due to the selective pressure to which it is submitted. PMID- 9335153 TI - Ultrastructural localization of the secretory aspartyl proteinase in Candida albicans cell wall in vitro and in experimentally infected rat vagina. AB - Detection and ultrastructural localization of aspartyl proteinase (Sap) in Candida albicans experimentally infecting rat vagina were studied. Two Sap positive (Sap+) and one Sap-negative (Sap-) strains of the fungus, endowed with high and low experimental vaginopathic potential, respectively, were used. Both Sap+ strains produced consistent Sap levels in the rat vagina, while the Sap- strain did not produce any measurable Sap. Electron microscopy of thin sections of chemically-fixed vaginal scrapings showed clear evidence of hyphae of proteolitic strains of C. albicans invading the keratinized epithelial cell layer of the vagina. The fungal cells exhibited a pronounced fibrillar layer on the cell wall with a marked intermixing of fungal and vaginal materials especially pronounced at the hyphal tip. Post-embedding immunogold techniques with the use of anti-Sap polyclonal and the specifically generated monoclonal antibody GF1 showed that Sap was essentially localized in the cell wall of C. albicans early during infection, in a cytological pattern mirroring Sap localization in C. albicans cells grown in Sap-inductive media in vitro. In summary, the data offer a new biochemical and ultrastructural evidence that Sap is actively secreted during experimental rat vaginitis by C. albicans. Cell wall localization of Sap is probably inherent to this active secretion process. PMID- 9335154 TI - Dermatophytes isolated from domestic animals in Barcelona, Spain. AB - This retrospective study deals with the main samples studied at the Mycology Diagnostic Service of the Faculty of Veterinary Science of Barcelona: animals with suspected dermatophytosis. Over a ten-year period from 1986 to 1995, 136 dermatophytes were identified from dog and cat cultures submitted for identification and from specimens submitted for mycological examination from a variety of other domestic animals. The most frequent dermatophytes isolated were Microsporum canis (55.9%), Trichophyton mentagrophytes var. mentagrophytes (27.2%), Microsporum gypseum (7.4%) and Trichophyton verrucosum (7.4%). The identity of the dermatophytes from dog and cat cultures submitted for identification was M. canis (77.8%), T. mentagrophytes (13.3%) and M. gypseum (8.9%). Dermatophytes were cultured from 15 of 105 (14.3%) canine specimens and 19 of 56 (33.9%) feline specimens submitted for mycological examination during this period. Microsporum canis was the most common species isolated (73.3% and 94.7% respectively). The percentage of positive microscopic examinations of the specimens of hair in culture positive submissions from dogs and cats was 58.8%. There was a high proportion of positive cultures from both dogs and cats less than 1 year of age, and in some breeds of dogs, but there was no significant difference between the sexes. Although dermatophytes were more frequently isolated in autumn and winter months, no significant difference was detected in the seasonal distribution of the canine and feline dermatophytosis. Trichophyton mentagrophytes was the most prevalent dermatophyte in rabbits and T. verrucosum in ruminants. Other isolated species were T. equinum and M. equinum from horses. PMID- 9335158 TI - Methylmercury in fish from Owyhee Reservoir in southeast Oregon: scientific uncertainty and fish advisories. AB - Data collected during 1987-1994 showed elevated levels of mercury (Hg) in fish tissue from the Owyhee Reservoir in southeastern Oregon. Sixty-five percent of the samples analyzed had total Hg levels exceeding the US Environmental Protection Agency's (EPA) health screening value of 0.6 mg/kg. Eighteen out of 89 (20%) fish tissue samples also had total Hg levels greater than the US Food and Drug Administration's (FDA) mercury action level of 1.0 mg/kg. The overall mean Hg content for all fish collected from the reservoir was 0.75 mg/kg wet weight (wet wt.). Fish muscle taken from largemouth bass (Micropterus salmoides), smallmouth bass (Micropterus dolomieu) and channel catfish (Ictalurus punctatus) had the highest mean Hg levels of 0.92, 0.87 and 0.82 mg/kg, respectively. In contrast, rainbow trout (Salmo gairdneri) had the lowest mean Hg content of 0.37 mg/kg. Increases in total Hg concentrations were found to be positively correlated with size for rainbow trout and yellow perch. A weak but significant correlation was also observed between total mercury content and age for smallmouth bass. Based on these data, in 1994 the Oregon Health Division (OHD) issued a fish consumption advisory for the Owyhee Reservoir using a conservative risk-based approach. The process of defining and communicating these consumption limits is the subject of this paper. PMID- 9335159 TI - Heavy metals input with phosphate fertilizers used in Argentina. AB - Sustainability of conventional agriculture is based upon a high input of agrochemicals, such as phosphate fertilizers. Conventional inorganic phosphorus fertilizers may cause an inadvertent addition of heavy metals, which are contained as impurities. Fertilizers commonly used in Argentina were analyzed to determine concentrations of chromium, cadmium, copper, zinc, nickel and lead. Rock phosphate contained the highest levels of cadmium and zinc, chromium was enhanced in diammonium phosphate and copper and lead were high in one superphosphate sample. Urea-phosphate contained the lowest levels of heavy metals. Concentrations of heavy metals varied considerably and the levels of Cd and Pb in some analyzed materials were significant relative to those naturally present in soils. Continuous fertilization of soils could increase the heavy metal contents exceeding natural abundances in soils, and transfer of these metals to the human food chain must not be overlooked. PMID- 9335160 TI - Population-based biomonitoring in the Czech Republic--the system and selected results. AB - In the framework of the system of monitoring the environmental impact on population health, the concentration of lead, cadmium and selenium in blood and cadmium in urine was measured in adults (n = 670), children (n = 599) and umbilical blood (n = 549) using atomic absorption spectrophotometry. Furthermore, cytogenetic analysis of peripheral lymphocytes in all population groups under study was investigated. The median blood Pb level for the overall group of adults (47.8 micrograms/l, i.e. 0.23 mumol/1) was significantly higher in men (51.5 micrograms/l, i.e. 0.25 mumol/l). Smoking significantly influenced the blood Pb level in women. The 90th percentile in no group exceeded the value of 100 micrograms/l (0.48 mumol/l). The median blood Cd level in adults (0.9 microgram/l, i.e. 0.008 mumol/l) depends on smoking habit (1.25 micrograms/l, i.e. 0.01 mumol/l). The median urine Cd level was 0.585 microgram/g creatinine (0.59 mumol/mole creatinine) in adults and 0.37 microgram/g creatinine (0.37 mumol/mole creatinine) in children. The median blood Se level (53.5 micrograms/l, i.e. 0.68 mumol/l) was found to be higher in the group of non-smokers (57.5 micrograms/l, i.e 0.73 mumol/l). Lead and selenium level were significantly lower in the umbilical blood. Cytogenetic analysis results showed age-dependent average percentages of aberrant cells: 1.1% in umbilical blood, 1.27% in children and 1.71 in adults in line with the reference values for the Czech population. PMID- 9335161 TI - Microscopic FTIR studies of lung cancer cells in pleural fluid. AB - Structural changes associated with lung cancer and tuberculous cells in pleural fluid were studied by microscopic FTIR spectroscopy. Infrared spectra demonstrate significant spectral differences between normal, lung cancer and tuberculous cells. The ratio of the peak intensities of the 1030 and 1080 cm-1 bands (originated mainly in glycogen and phosphodiester groups of nucleic acids) differs greatly between normal and lung cancer samples. Such findings prompt the consideration that recording infrared spectra from lung cancer and tuberculous cells may be of diagnostic value. Since measurements of IR spectra of lung cancer cells in the pleural fluid can be a very rapid inexpensive process, our finding warrant exploration of this possibility in the investigation of the mechanism whereby the environmental pollution related cancers develop. PMID- 9335162 TI - Semen quality in workers with long-term lead exposure: a preliminary study in Taiwan. AB - The objective of this study was to evaluate the semen quality from five workers in a lead battery manufacturing factory. Blood and semen samples were taken to measure lead concentrations. These samples were compared with samples from eight workers not exposed to lead, based on age, marital status and duration of marriage. Results showed lead concentrations in blood and semen were significantly higher in the exposed group. However, comparisons based on sperm count, motility and viscosity showed no significant differences. There was a significant negative relationship for the exposed group between motility and blood-lead and semen-lead concentrations. Comparing sperm count and blood-lead and semen-lead concentrations, there was a non-significant negative relationship. Semen-lead concentrations and semen pH were significant factors that explained the semen count and semen motility results. Prolonged exposure to lead may have harmful effects on the body's reproductive functions. PMID- 9335163 TI - The cellulolytic system of Clostridium cellulolyticum. AB - Recent findings on the cellulolytic system of the mesophilic Clostridium cellulolyticum are reviewed. Six cellulases and the scaffolding protein, which are, at the present time, the known components of the cellulosome have been cloned. The catalytic and structural properties of the cloned enzymes CelA, CelC, CelD and CelF are described. It was shown that the grafting of the cellulases onto the scaffolding protein was performed using the dockerin-cohesin attachment device and was strictly dependent on the integrity of both components of the complex. The amino-acid sequences of dockerin and cohesin domains of C. cellulolyticum were compared to that of C. cellulovorans and C. thermocellum. This sequence analysis shows that domains belonging to the thermophilic or the mesophilic bacteria can be placed into two well defined groups. The genetic organization of the gene cluster of C. cellulolyticum is discussed. PMID- 9335164 TI - Synergism between the cellulosome-integrating protein CipA and endoglucanase CelD of Clostridium thermocellum. AB - The cellulosome-integrating protein CipA of Clostridium thermocellum was produced from a recombinant clone of Escherichia coli and purified by cellulose affinity and anion exchange chromatography. Two active forms of C. thermocellum endoglucanase CelD were tested for binding and hydrolytic activity on Avicel in the presence and in the absence of CipA. One was 68 kDa CelD, which contains an intact dockerin domain. The other was 65 kDa CelD, in which the dockerin domain is partially deleted. CipA promoted quantitative binding of 68 kDa CelD to Avicel and enhanced its Avicelase activity by at least ten-fold. By contrast, CipA had no effect on the activity nor on the cellulose-binding affinity of the truncated 65 kDa form. These results show that interaction between CipA and the catalytic component CelD is needed for the degradation of cellulose and confirm that the interaction is mediated by the dockerin domain of CelD. PMID- 9335165 TI - Trichoderma reesei cellobiohydrolase I with an endoglucanase cellulose-binding domain: action on bacterial microcrystalline cellulose. AB - Cellulolytic enzymes consist of distinct catalytic and cellulose-binding domains (CBDs). The presence of a CBD improves the binding and activity of cellulases on insoluble substrates but has no influence on their activities on soluble substrates. Structural and biochemical studies of a fungal CBD from Trichoderma reesei cellobiohydrolase I have revealed a wedge shaped structure with a flat cellulose binding surface containing three essential tyrosine residues. The face of the wedge is strictly conserved in all fungal CBDs while many differences occur on the other face of the wedge. Here we have studied the importance of these differences on the function of the T. reesei CBHI by replacing its CBD by a homologous CBD from the endoglucanase, EGI. Our data shows that, apart from slightly improved affinity of the hybrid enzyme, the domain exchange does not significantly influence the function of CBHI. PMID- 9335166 TI - Cellulose binding domains and linker sequences potentiate the activity of hemicellulases against complex substrates. AB - To evaluate the role of the CBDs and linker sequences in Pseudomonas xylanase A (XYLA) and arabinofuranosidase C (XYLC), the catalytic activity of derivatives of these enzymes, lacking either the linker sequences or CBDs, was assessed. Removal of the CBDs or linker sequences did not affect the activity of either XYLA or XYLC against soluble arabinoxylan, while derivatives of XYLA, in which either the CBD or interdomain regions had been deleted, exhibited decreased activity against the xylan component of cellulose/hemicellulose complexes. Although a truncated derivative of XYLC (XYLC"'), lacking its CBD, was less active than the full length enzyme against plant cell wall material containing highly substituted arabinoxylan, XYLC"' was more active than XYLC on complex substrates where the degree of substitution of arabinoxylan was very low. These data indicate that CBDs and linker sequences play an important role in the activity of hemicellulases against plant cell walls and other cellulose/hemicellulose complexes. PMID- 9335167 TI - Enzymatic properties of cellulases from Humicola insolens. AB - We present the analysis of the activities towards soluble and insoluble substrates of seven cellulases cloned from the saprophytic fungus Humicola insolens. The activity on the soluble polymer substrate carboxymethylcellulose (CMC) was used to determine the pH activity profiles of the five endoglucanases (EG), whereas cellotriose and reduced cellohexaose were used to determine the pH activity profiles of cellobiohydrolase I (CBH) and CBH II. All the EGs show optimal activity between pH 7 and 8.5, while CBH I and CBH II peak around pH 5.5 and 9, respectively. The catalytic activities of five of these cellulases were investigated under neutral and alkaline conditions using reduced cellohexaose as a substrate in a cellobiose oxidase coupled assay. EG I and CBH I both belong to family (7) according to a recent classification of glycosyl hydrolases. They both have activity against cellotriose. Therefore, they were studied using a coupled assay involving glucose oxidase. The activity on insoluble substrate (phosphoric acid swollen cellulose) was assessed by the formation of reducing groups. The presence of a cellulose binding domain (CBD) lowers the apparent KM. This can be explained by the dispersing action of CBD. However, the CBD also reduces the apparent k(cat) probably by slowing down the mobility. EG I, EG II and EG III show similar activity towards CMC and amorphous cellulose, while EG V, EG VI, CBH I and CBH II have the highest catalytic rate on amorphous cellulose. In summary, Humicola insolens possesses a battery of cellulose-degrading enzymes which cooperate in the efficient hydrolysis of cellulose. PMID- 9335168 TI - Oligosaccharide specificity of a family 7 endoglucanase: insertion of potential sugar-binding subsites. AB - Family 7 of the glycosyl hydrolases contains both endoglucanases and cellobiohydrolases. In addition to their different catalytic activities on crystalline substrates, the cellobiohydrolases differ from the endoglucanases in their activity on longer soluble substrates, indicative of a greater number of subsites on the enzyme. A double mutant (S37W, P39W) of the Humicola insolens endoglucanase I (EG I) has been constructed in order to mimic aspects of the subsite structure of the corresponding family 7 cellobiohydrolase, cellobiohydrolase-I (CBH I). The 3-D crystal structure of the double mutant has been solved and refined to a crystallographic R-factor of 0.17 at a resolution of 2.2 A (1 A = 0.1 nm). The two mutant tryptophans are clearly visible in the electron density and are in the same orientation as those found in the substrate binding groove of CBH I. In addition to the substitutions, the C-terminal amino acids (399QELQ), disordered in the native enzyme structure, are clearly visible and there are a small number of minor loop movements associated with differences in crystal packing. Kinetic determinations show that the S37W, P39W mutant EG I has almost identical activity, compared to native EG I, on small soluble cellodextrins. On phosphoric acid swollen cellulose there is a small (30%), but significant, decrease in the apparent KM indicating that the double mutant may indeed exhibit stronger binding to longer polymeric substrates. PMID- 9335169 TI - Surface residue mutations which change the substrate specificity of Thermomonospora fusca endoglucanase E2. AB - The three dimensional structure of a T. fusca endoglucanase catalytic domain (E2cd) has been determined by X-ray crystallography at 1.0 A resolution (Wilson et al., 1995). The availability of a high resolution structure for E2cd allows us to initiate structure-based efforts to engineer cellulases with a high activity on native cellulose. The low activity on crystalline cellulose suggests that the entry of a cellulose molecule into the active site rather than catalysis may be the rate limiting step for hydrolysis of crystalline cellulose. Movement of a loop upon substrate binding has been proposed to play a crucial role in catalysis. A total of 15 surface mutants and 5 loop mutants were created by site directed mutagenesis and their effects on activity and substrate specificity were determined. Circular dichroism spectra were used to monitor structural changes, and no major changes were found. The binding constants for two methyl umbelliferyl oligosaccharides and cellotriose were measured for some of the mutants and all of them showed binding similar to wild type E2. These results provide the first direct link between loop movement and catalysis by E2 and show that surface residues can affect its substrate specificity. PMID- 9335170 TI - Structural and functional properties of low molecular weight endo-1,4-beta xylanases. AB - There are currently four crystal structures of low molecular weight endo-1,4-beta xylanases (E.C.3.2.1.8), i.e. family G/11 xylanases, available at the Brookhaven Data Bank: 2 xylanases from Trichoderma reesei (Torronen et al., 1994; Torronen and Rouvinen, 1995) and one from Bacillus circulans and another from Trichoderma harzianum (Campbell et al., 1993). They consist of two beta-sheets and one alpha helix and have been described to resemble a partly-closed right hand. The catalytic residues are two conserved glutamate residues, which are located opposite to each other in an open active site cleft. The catalytic mechanism is thought to resemble that of the widely-studied enzyme lysozyme. The role of one glutamate is to act as an acid/base catalyst whereas the other is a nucleophile and stabilizes the reaction intermediate. Complex structures of partly-bound xylotetraose in mutated XYN from Bacillus circulans (Wakarchuck et al., 1994a) and three recently-obtained structures of XYNII from Trichoderma reesei with epoxyalkyl-xylose derivatives (Havukainen et al., 1996) have provided important information on substrate binding. Family G/11 xylanases show clear amino acid homology and thus have a common fold. However, variations in their functional properties, such as catalytic activity, substrate cleaving patterns, pH optima and thermostabilities, exist. PMID- 9335171 TI - Endo-beta-1,4-xylanase families: differences in catalytic properties. AB - Microbial endo-beta-1,4-xylanases (EXs, EC 3.2.1.8) belonging to glycanase families 10 (formerly F) and 11 (formerly G) differ in their action on 4-O-methyl D-glucurono-D-xylan and rhodymenan, a beta-1,3-beta-1,4-xylan. Two high molecular mass EXs (family 10), the Cryptococcus albidus EX and XlnA of Streptomyces lividans, liberate from glucuronoxylan aldotetrauronic acid as the shortest acidic fragment, and from rhodymenan an isomeric xylotriose of the structure Xyl beta 1-3Xyl beta 1-4Xyl as the shortest fragment containing a beta-1,3-linkage. Low molecular mass EXs (family 11), such as the Trichoderma reesei enzymes and XlnB and XlnC of S. lividans, liberate from glucuronoxylan an aldopentauronic acid as the shortest fragment, and from rhodymenan an isomeric xylotetraose as the shortest fragment containing a beta-1,3-linkage. The structure of the oligosaccharides was established by: NMR spectroscopy, mass spectrometry of per-O methylated compounds and enzymic hydrolysis by beta-xylosidase and EX, followed by analysis of products by chromatography. The structures of the fragments define in the polysaccharides the linkages attacked and non-attacked by the enzymes. EXs of family 10 require a lower number of unsubstituted consecutive beta-1,4 xylopyranosyl units in the main chain and a lower number of consecutive beta-1,4 xylopyranosyl linkages in rhodymenan than EXs of family 11. These results, together with a greater catalytic versatility of EXs of family 10, suggest that EXs of family 10 have substrate binding sites smaller than those of EXs of family 11. This suggestion is in agreement with the finding that EXs of family 10 show higher affinity for shorter linear beta-1,4-xylooligosaccharides than EXs of family 11. The results are discussed with relevant literature data to understand better the structure-function relationship in this group of glycanases. PMID- 9335172 TI - Definition of the substrate specificity of the 'sensing' xylanase of Streptomyces cyaneus using xylooligosaccharide and cellooligosaccharide glycosides of 3,4 dinitrophenol. AB - The title compounds, (Xylp beta (1-->4))nXylp beta-3,4-DNP (n = 0-4) have been made by selective anomeric deprotection of peracetylated xylose oligosaccharides with hydrazine, followed by formation of the trichloroacetimidate, uncatalysed reaction with 3,4-dinitrophenol, and Zemplen deacetylation. The values of k(cat)/K(m) for 3,4-dinitrophenol release from these substrates by xylanase III of Streptomyces cyaneus, expressed in Escherichia coli, increase with increasing n up to n = 2 and then slightly decrease. Since it is known from previous work that in its normal host, the enzyme is produced constitutively at low levels and excreted, these results suggest that the biological function of the enzyme may be to produce small molecule inducers, predominantly xylotriose, from the non reducing end of the xylan. Activity on cellooligosaccharide glycosides (Glcp beta (1-->4))nGlcp beta-3,4-DNP (n = 0-3) was detected, at a rate about two-and-a-half orders of magnitude less than that observed on the corresponding xylooligosaccharides, indicating that the enzyme is a true xylanase. PMID- 9335173 TI - Action of Trichoderma reesei mannanase on galactoglucomannan in pine kraft pulp. AB - The di-, tri- and tetrasaccharides formed during Trichoderma reesei endo-beta-D mannanase treatment of pine kraft pulp were studied. The oligosaccharides in the hydrolysate were fractionated using size-exclusion, anion exchange and activated carbon chromatography. The primary sequence of the purified oligomers was determined by two-dimensional NMR techniques. The T. reesei mannanase cleaves the beta-1,4-glycosidic linkage of D-mannosyl residues attached either to D-mannose or D-glucose. The D-mannosyl residue may also be substituted by a D-galactosyl group. The main disaccharide produced was mannobiose, but a significant amount of 4-O-beta-D-glucopyranosyl-D-mannopyranose (GlcMan) was also produced. After extensive hydrolysis the main trisaccharides produced were 4-O-beta-D mannopyranosyl-[6-O-alpha-galactopyranosyl]-D-mannopyranose (Gal1Man2) and 4-O beta-D-glucopyranosyl-4-O-beta-D-glucopyranosyl-D-mannopyranose (Glc2Man). Some mannotriose 4-O-beta-D-glucopyranosyl-4-O-beta-D-mannopyra-nosyl-D-manno pyranose (GlcMan2) and 4-O-beta-D-glucopyranosyl-[6-O-alpha-galactopyranosyl]-D mannopyranose (Gal1GlcMan) were also detected in the hydrolysate. The structures of two tetrasaccharides were studied. They appeared to be 4-O-beta-D glucopyranosyl-4-O-beta-D-glucopyranosyl-4-O-beta-D- glucopyranosyl-D mannopyranose (Glc3Man) and 4-O-beta-D-glucopyranosyl-4-O-beta-D-mannopyranosyl-4 O-beta-D -glucopyranosyl-D-mannopyranose (GlcManGlcMan). According to the results obtained, the galactoglucomannan in pine contains regions in which two or three glucose units are linked together, which further means that it may contain regions with several successive mannose residues. The galactose side groups were found to be attached only to mannose. PMID- 9335174 TI - Factors determining more efficient large-scale release of a periplasmic enzyme from E. coli using lysozyme. AB - Large scale use of lysozyme for periplasmic release has been impeded by the cost of the pure enzyme and its subsequent presence as a contaminant in later downstream processing steps. In this paper, we discuss the use of lysozyme for pilot scale recovery of a periplasmic enzyme from E. coli. The effects of concentration of sucrose, lysozyme and cells on periplasmic enzyme release were examined. Lysozyme concentration can be reduced 5-fold from previous reports and a reduction in sucrose concentration from 20 to 15% (w/v) allows an improvement in centrifugal harvesting by reducing viscosity. High levels of release were still achieved using this technique and further improvements in yield were obtained by optimising other components of the releasing mixture. Results show that some release is still achieved in circumstances where no lysozyme use is possible. Results also indicate that a substantial proportion (up to 70%) of lysozyme remains bound to the cellular debris after its action and is removed with this material. PMID- 9335175 TI - Overexpression of the Aspergillus niger pH 2.5 acid phosphatase gene in a heterologous host Trichoderma reesei. AB - An Aspergillus gene coding for a pH 2.5 acid phosphatase enzyme was successfully overexpressed in Trichoderma reesei under the strong main cellobiohydrolase I (cbh 1) promoter. The best transformants produced up to 240 times more of the acid phosphatase than the Aspergillus strain from which the phosphatase gene was originally isolated. The recombinant enzyme was effectively secreted into the culture medium both by its own and the cbh 1 secretion signal. The heterologous pH 2.5 acid phosphatase enzyme produced by the Trichoderma transformants was seen as four protein bands of about 55-66 kD resulting from variable glycosylation in Trichoderma. The activity of the recombinant enzyme was not affected. Enzyme preparations rich in both cellulose and phytate hydrolysing enzymes are of interest in the animal feed industry. PMID- 9335176 TI - Influence of surface hydrophilic/hydrophobic balance on enzyme properties. AB - Two different enzyme surface modifications were carried out in order to alter the protein hydrophilic/hydrophobic balance in opposite directions and to observe the effects induced on enzyme properties. First, a novel chemoenzymatic glycosylation method was applied, which resulted in a higher enzyme surface hydrophilic character. Then, an amphiphilic polymer, PEG, was bound to the enzymes by chemical means, and it brought about an increase in the global hydrophobic character. Two different enzymes, alpha-chymotrypsin and Candida rugosa lipase, were studied, and in all cases, several degrees of modification were obtained. Then, the modified biocatalysts were thoroughly investigated, and the influence of the variation of surface hydrophilic/hydrophobic balance on hydrolytic activity, hydrolysis kinetic parameters, synthetic activity and thermal stability was assessed. PMID- 9335177 TI - Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) in recombinant Escherichia coli grown on glucose. AB - A recombinant Escherichia coli strain has been developed that produces poly(3 hydroxybutyrate-co-4-hydroxybutyrate) when grown in complex medium containing glucose. This has been accomplished by introducing into E. coli DH5 alpha separate plasmids harboring the polyhydroxyalkanoate (PHA) biosynthesis genes from Ralstonia eutropha (formerly named Alcaligenes eutrophus) and the succinate degradation genes from Clostridium kluyveri, respectively. Poly(3-hydroxybutyrate co-4-hydroxybutyrate) levels reached 50% of the cell dry weight and contained up to 2.8 mol.% 4-hydroxybutyrate. The molecular weight of the polymer was 1.8 x 10(6). PMID- 9335178 TI - Overproduction and purification of an agarase of bacterial origin. AB - The agarase gene from Streptomyces coelicolor has been cloned in the non-producer bacterium Streptomyces lividans under the control of its own set of promoters and under the control of a heterologous promoter that is functional only during exponential growth. The best level of overproduction was obtained when the strain containing the natural gene was cultivated in fed batch with mannitol as carbon source. The protein, with a relative molecular mass of 32 kDa, has been purified following an affinity purification method. Contaminating activities seem to be absent from the purified enzyme preparation that can be used to purify DNA from agarose gels. PMID- 9335179 TI - Development of mass rearing technique of Tyrophagus putrescentiae (Acari: Acaridae) found in house dust. AB - A storage mite, Tyrophagus putrescentiae, is recently known to be widely distributed in Korea, being commonly found in house dust, and may, therefore, be allergenically important. The purpose of this study was to develop mass rearing techniques for supplying a large quantity of allergens. The laboratory mouse food powder gave the highest yield, showing 1,251.5-fold increase in number after 10 weeks, and the mixed powder of laboratory mouse food and yeast (1:1) also gave same level of the production (1,203.1-fold increase in week 10). Several different combinations of temperature and relative humidity conditions were compared, and the maximum propagation was obtained at 25 degrees C and 64% RH, showing 960-fold increase in number. When the same amount of culture media was used, the size of the culture container did not significantly influence the quantitative yield of T. putrescentiae mites. PMID- 9335180 TI - Changing patterns of Clonorchis sinensis infections in Kyongbuk, Korea. AB - Studies were conducted from May, 1993 to April, 1995 to determine the changing patterns of infection by the liver fluke, Clonorchis sinensis, among residents and fish hosts in Kyongbuk Province. The infection rate among residents was 7.7% by stool examination. The rate in males (11.3%) was significantly higher than females (4.1%). Positive rate of intradermal test was 27.6% in the same population. The special type of a simple catalytic model was applied for the analysis of intradermal positive reactors by age and sex, and the equation was y = 0.4776 (1 - e-0.0375t) for males and, y = 0.2085 (1 - e-0.0138t) for females. Analysis of stool examination data by two-stage catalytic model revealed y = 0.025 (e-0.00471 - e-0.0235t). The annual Clonorchis infection rate was 4.7 per 1,000 susceptibles and the annual loss rate was 23.5 per 1,000 infected. The frequency distribution by the eggs per gram (EPG) was calculated as well as the cumulative percentages of positives. The regression equations were y = 0.929 + 1.506 log x for males and, y = 0.473 + 1.767 log x for females. Of the 25 fish species, 7 species were infected with Clonorchis metacercariae. Infection rates varied by the species, and ranged from 2.8% in Puntungia herzi to 30.0% in Pseudorasbora parva. Average number of the matacercariae per gram of flesh was 58.1 in P. parva, followed by 10.2 in Gnathopogon atromaculatus, 7.0 in Saurogobio dabryi, and 3.0 in Paracheilognathus rhombea. The present study indicates that clonorchiasis in Kyongbuk Province is less prevalent than that of several decades ago. PMID- 9335181 TI - [Infection status with trematode metacercariae in the fresh-water fish from Chunamchosuchi (pond), Uichang-gun, Kyongsangnam-do, Korea]. AB - The present study was performed to analyze the infection status of trematode metacercariae in fishes caught from Chunamchosuchi (pond) located in Uichang-gun, Kyongsangnam-do. A total of 130 freshwater fish of 5 species was collected by a fish net and fish traps from November, 1995 to May, 1996. They were examined under a stereomicroscope after artificial digestion with pepsin-HCl solution. A total of 8 species of metacercaria, i.e. Clonorchis sinensis, Echinochasmus japonicus, Cyathocotyle orientalis, Diplostomum sp., Metorchis orientalis, Holostephanus nipponicus, Exorchis oviformis and unidentified echinostome, was detected from them. The metacercariae of C. sinenesis were found in 8/20 (40.0%) Acanthorhodeus asmussi, 20/20 (100%) Culter brevicauda, 31/45 (68.9) Cultriculus eigenmanni and 21/25 (84.0%) Pseudorasbora parva, and the average number of metacercariae detected in each fish species were 1.9, 31.7, 15.3, and 73.0. From the above results, it was confirmed that fresh-water fishes from Chunamchosuchi (pond) were highly infected with metacercariae of avian trematode, i.e. C. orientalis, H. nipponicus, M. orientalis, E. japonicus and Diplostomum sp., and 4 species of fish, P. parva, C. brevicauda, C. eigenmanni and A. asmussi, were infected with metacaecariae of C. sinensis. PMID- 9335183 TI - Effects of Cryptosporidium baileyi infection on the bursa of Fabricius in chickens. AB - In order to clarify the effect of cryptosporidiosis on immune response, histopathological changes associated with experimentally occurring bursal cryptosporidiosis in chickens were chronologically observed as the first step. A total of 150 2-day-old chickens was each inoculated orally with a single dose of 5 x 10(5) Cryptosporidium baileyi oocysts. The chickens showed a normal profile of oocyst shedding in droppings. The bursa indices throughout the experimental period indicated negligible reactions. Numerous cryptosporidia occurred in the microvillous border of bursal epithelium between days 4 and 16 postinoculation (PI). Appearance of the most mast cells was followed by a dramatic loss of the protozoa in the bursa of Fabricius (BF). The distribution of the coccidium coincided with heterophil infiltration in the epithelium and adjacent lamina propria. The histopathological lesion was marked diffuse chronic superficial purulent bursitis with heterophil infiltration in the epithelium and adjacent lamina propria and mucosal epithelial hyperplasia. These results suggest that the bursitis may induce immunosuppressive effect. PMID- 9335182 TI - Geographical distribution of vectors and sero-strains of tsutsugamushi disease at mid-south inland of Korea. AB - Studies of geographical distributions and relative population densities of the vector mites of tsutsugamushi disease were carried out in October 1996 at 12 locations of the mid-south inland of the Korean peninsula, where chigger mites have been never studied. Of 177 field rodents and insectivores collected, 154 (87.0%) were Apodemus agrarius. Total 25,707 chigger mites were collected and 14 species were identified, of which Leptotrombidium pallidum was predominant (79.8%) and L. palpale the next (8.9%). L. pallidum, the vector species, was widely distributed in all study areas, showing the highest density at Cho-o 2 dong, Sangju-si (chigger index 201.8), and the lowest at Tanwol-dong. Chungju-si (chigger index 40.7). The other vector species, L. scutellare was found only at the southern part of the study area such as Yobae and Mipyong, Kumrunggun and Unsu, Kimchon-si. The northernmost areas of the L. scutellare distribution were coincided with the areas where anual mean air temperature is above 10.0 C. Among 157 A. agrarius sera tested, 48.3% was Karp, 1.7% Gilliam and 3.3% Kuroki. The rest of the sera were not able to determine the sero-type because of the cross antigen-antibody reactions among the tested sero-types. PMID- 9335184 TI - The role of nitric oxide as an effector of macrophage-mediated cytotoxicity against Trichomonas vaginalis. AB - The purpose of this study is to determine whether nitric oxide is involved in the extracellular killing of Trichomonas vaginalis by mouse (BALB/c) peritoneal macrophages and RAW264.7 cells activated with LPS or rIFN-gamma and also to observe the effects of various chemicals which affect the production of reactive nitrogen intermediates (RNI) in the cytotoxicity against T. vaginalis. The cytotoxicity was measured by counting the release of [3H]-thymidine from labelled protozoa and NO2- was assayed by Griess reaction. NG-monomethyl-L-arginine (L NMMA), NG-nitro-L-arginine methyl ester (NAME) and arginase inhibited cytotoxicity to T. vaginalis and nitrite production by activated mouse perioneal macrophages and RAW 264.7 cells. The addition of excess L-arginine competitively restored trichomonacidal activity of macrophages. Exogenous addition of FeSO4 inhibited cytotoxicity to T. vaginalis and nitric products of macrophages. From above results, it is assumed that nitric oxide plays an important role in the host defense mechanism of macrophages against T. vaginalis. PMID- 9335185 TI - Variation of antigenic proteins of eggs and developmental stages of Paragonimus westermani. AB - Diagnosis of early paragonimiasis is difficult because parasitological evidence is not easily obtained. Antibody tests have been proposed as a good substitute for classical diagnostic techniques. Using the crude extracts of Paragonimus westermani eggs, metacercariae, 4- and 7-week juveniles, and 16-week adults as antigens, we observed the early antibody responses. Sera were obtained from 4 experimental cats, fed 50 metacercariae each, at intervals until 13 weeks post infection. Antibody (IgG) responses were identified by ELISA using extracts of 4 week juveniles, followed by those of 7- and 16-week worms. Antibody responses were minimal against the metacercarial extracts. Antibodies to P. westermani egg extracts were elevated after 10 weeks post-infection. In immunoblot analysis, more than nine protein bands in 4-week juveniles reacted with the early infection sera. Antigenic proteins in adult worms were different from those of juveniles. After four weeks of infection, 32 and 35 kDa bands in the adult extracts were increasingly reactive. Egg specific proteins at 28, 46 and 94 kDa were reactive only after 10 weeks. Antigenic components reacting to the early infection sera changed during the maturation stages of P. westermani; almost all juvenile antigens were replaced by adult antigen components. PMID- 9335186 TI - [cDNAs encoding the antigenic proteins in pathogenic strain of Entamoeba histolytica]. AB - The differential display reverse transcription polymerase chain reaction (DDRT PCR) analysis was performed to identify the pathogenic strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenic strain YS-27 and the non-pathogenic strain S 16, respectively. Three kinds of first stranded cDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT11M (M: A, C, or G) primers. Each cDNA template was used for DDRT-PCR analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oligo dT11M primers. Of these, 31 amplicons were verified as the amplicons amplified only from the mRNAs of the pathogenic strain by DNA slot blot hybridization. Further characterization of the 31 pathogenic strain specific amplicons by DNA slot blot hybridization analysis using biotin labeled probes of the PCR amplified DNA of cysteine proteinase genes revealed that 21 of them were amplified from the mRNAs of the cysteine proteinase genes. Four randomly selected amplicons out of the rest 10 amplicons were used for screening of cDNA library followed by immunoscreening and all of them were turned out to be amplified from the mRNA. PMID- 9335187 TI - Susceptibility of some vertebrate hosts to infection with early third-stage larvae of Gnathostoma hispidum. AB - Susceptibility of some vertebrates was examined to the early third-stage larvae (EL3) of Gnathostoma hispidum. The larvae collected from the Chinese loaches were infected to 4 silk carps, 3 snake heads, 3 bullfrogs, 5 mice and 9 albino rats. No worms were detected in fish, silk carps and snake heads. In 3 bullfrogs fed 30 larvae, a total of 9 EL3 was recovered in the gastrointestinal tract (8 larvae) and liver (one). In 5 mice infected with 50 larvae, a total of 37 (74.0%) advanced third-stage larvae (AdL3) was recovered from the muscle (31 larvae), liver (5 larvae) and kidney at 4 weeks after infection. In 9 albino rats infected with 115 larvae, a total of 40 (34.8%) AdL3 was found in the muscle. The mammalian hosts were found susceptible to the EL3 of G. hispidum from Chinese loaches. PMID- 9335188 TI - An unusual over-gravid female of Enterobius vermicularis recovered from a child. AB - An unusual over-gravid female of Enterobius vermicularis was recovered from a 15 month old child by cello-tape anal swab. The patient resided in Inchon and complained of severe anal itching. The worm measured 7.8 mm in length and 0.5 mm in width, and retained typical morphologic features of E. vermicularis such as cephalic alae and a sharply pointed posterior end. In this gravid female, peculiarly, the uterus was tremendously distended, and about 99% of the whole body length was completely packed with a great number of eggs. Other internal organs were difficult to observe. This paper describes a peculiar over-gravid female of E. vermicularis. PMID- 9335190 TI - Dracunculiasis and onchocerciasis. PMID- 9335191 TI - Expanded programme on immunization (EPI). PMID- 9335189 TI - A rare association between ovarian endometriosis and bilateral ovarian teratoma. Case report. AB - A 28 year woman complained for a sudden abdominal pain with some of the classical signs of the acute abdomen. Since a bilateral ovarian mass was found at ultrasound examination, clinician decided that patient underwent a urgent surgery. At histology, this mass resulted to be a bilateral triphillic mature teratoma. A focus lined by endometrioid epithelium and embedded in the modified stroma of endometrial type, was found beyond the residual cortex of the left ovary. There was some fibrotic tissue separating the endometrial and teratomatous lesions. To our knowledge, these two diseases have never been reported as associated in literature, but this association has a clinical relevance because an endometriotic pathology can reveal a silent teratoma with bilateral ovarian localization. PMID- 9335192 TI - The mobilisation of peripheral blood progenitor cells using chemotherapy, stem cell factor and filgrastim or chemotherapy plus filgrastim alone: a randomised study. PMID- 9335193 TI - Prediction versus management models relevant to risk assessment: the importance of legal decision-making context. AB - Most of the theoretical and empirical literature on violence risk to date has focused on the task of predicting who will behave violently. In the present article, it is argued that at least two models of risk assessment may be applied to the varying legal decisions in which violence risk is a consideration: prediction (with an emphasis on overall accuracy) and management (with an emphasis on risk reduction). These two models are described, and discussed in the contexts of the literatures on forensic assessment and therapeutic jurisprudence. The implications for research, policy, and practice are considered. PMID- 9335194 TI - The validity of mental patients' accounts of coercion-related behaviors in the hospital admission process. AB - Although the recent development of a measure for perceived coercion has led to great progress in research on coercion in psychiatric settings, there still exists no consensus on how to measure the existence of real coercive events or pressures. This article reports the development of a system for integrating chart review data and data from interviews with multiple participants in the decision for an individual to be admitted to a psychiatric hospital. The method generates a "most plausible factual account" (MPFA). We then compare this account with that of patients, admitting clinicians and other collateral informants in 171 cases. Patient accounts most closely approximate the MPFA on all but one of nine dimensions related to coercion. This may be due to wider knowledge of the events surrounding the admission. PMID- 9335195 TI - Forensic mental health clinical evaluation: an analysis of interstate and intersystemic differences. AB - Forensic mental health evaluation systems have undergone major changes during the past two decades, and the variability of service delivery systems across states is significant. We compared assessments of competence to stand trial and criminal responsibility in three states with different systems for forensic mental health evaluations: Michigan, Ohio, and Virginia. Although all three states use comparable legal criteria to judge competence and criminal responsibility, we found large, statistically significant differences among the states in the proportion of defendants referred for evaluation who were assessed as incompetent or not criminally responsible. In addition, significant differences were found in the diagnostic and offense categories of defendants referred for evaluation. Our findings suggest that the structure of a system for providing forensic evaluation services may significantly affect both the group of individuals referred for evaluation as well as evaluation outcome. PMID- 9335196 TI - The prevalence of mental illness among inmates in a rural state. AB - A limited number of recent empirical studies suggest that inmates suffer from high rates of serious mental illness. Different explanations are offered depending on the type of institution: jail or prison. The literature is based largely on urban samples and does not offer comparisons of rates across types of institution within a single study. The present study examined a random sample of 213 jail and prison inmates in a rural state using the Diagnostic Interview Schedule (III-R). Among jail inmates there was little evidence of high rates of serious mental illness, suggesting the criminalization of mental illness may not be as evident in rural settings as urban areas. Among prison inmates, however, high rates of mental disorders were found, supporting previous findings in urban and rural jurisdictions. Implications of the findings are discussed in the context of a consolidated correctional system. PMID- 9335197 TI - Using British trial procedures in American cases: a more civil trial? AB - A number of attorneys, judges, and legal scholars have asserted that the overly combative nature of American trials may impact on the actual quality of justice and bring the legal system into disrepute. In contrast, many who witness criminal and civil trials conducted in Great Britain are struck by the greater apparent civility of the courtroom atmosphere. Closer examination of the English system reveals seven specific procedural differences that may contribute to this perceived change in atmosphere. In this study, these procedural differences were manipulated and their effect on verdict and on perceptions of trial participants measured. In addition, opinions about these differences were elicited. Results showed that while the trial was perceived as more civil, and the judge viewed more positively, participants tended to indicate preferences for the American style. Implications of these results are discussed. PMID- 9335198 TI - Tyrosine hydroxylase allelic distribution in suicide attempters. AB - A tetranucleotide repeat polymorphism in the first intron of the tyrosine hydroxylase (TH) locus was examined in a group of 118 adult suicide attempters and 78 control subjects. The suicide attempters were diagnosed according to DSM III-R criteria at the index attempt and represented the following diagnoses: major depression (18), dysthymia (13), anxiety disorders (16), adjustment disorders (29), psychoactive substance abuse disorders (27) and psychotic disorders (15). A significant variation in the prevalence of carriers of the TH K3 allele (high for suicide attempters with adjustment disorders, P = 0.0023) and a tendency toward a variation of the TH-K1 allele (low among all suicide attempters, P = 0.046) was observed. In light of other data the variation of TH K1 and TH-K3 suggests that these alleles may reflect predisposition for a common phenotype with altered vulnerability for psychiatric disorders. PMID- 9335199 TI - Nocturnal secretion of prolactin and cortisol and the sleep EEG in patients with major endogenous depression during an acute episode and after full remission. AB - We investigated the sleep electroencephalogram (EEG) and the nocturnal secretion of prolactin and cortisol in 25 normal subjects and 12 male inpatients with major depression before treatment and after remission and drug withdrawal. In the depressed patients, sleep-EEG disturbances persisted after recovery, whereas the cortisol concentration decreased. Prolactin variables in the patients did not differ between the two time points (i.e. before treatment and after remission). Compared with the normal subjects, the patients had significantly higher cortisol concentrations. The above findings were not altered when age was used as a covariate in statistical analysis. Our data suggest that neither depression nor aging exerts distinct effects on prolactin secretion. PMID- 9335201 TI - Paranoid schizophrenia: non-specificity of neuropsychological vulnerability markers. AB - During stages of remission, patients with paranoid schizophrenia seldom show severe attentional or information-processing dysfunctions, except in cases of long-term chronicity. The diagnostic specificity of four putative psychological vulnerability indicators of schizophrenia - the Span of Apprehension, the degraded stimulus Continuous Performance Test (dsCPT), the degraded stimulus visual backward masking task and the Wisconsin Card Sorting Test (WCST) - was examined in a group of patients with paranoid schizophrenia. Since no single test seems to identify all patients, the use of a combination of measures may be a useful strategy. Accordingly, the four tests were administered to 18 paranoid schizophrenic patients, 18 depressed patients and 18 normal subjects. Paranoid schizophrenic patients could be distinguished from normal subjects primarily on the basis of their performance on the backward masking task and secondarily by the dsCPT and the WCST. Paranoid schizophrenic and depressed patients could be differentiated to some extent by their performance on an information-mask condition of the backward masking task. Thus, of the four measures studied, only the degraded stimulus backward masking appeared to be a specific indicator of paranoid schizophrenia. PMID- 9335200 TI - CSF 5-HIAA, testosterone, and sociosexual behaviors in free-ranging male rhesus macaques in the mating season. AB - This study examines sexual behavior, serotonin turnover in the central nervous system, and testosterone in free-ranging non-human primates. Study subjects were 33 young adult male rhesus macaques (Macaca mulatta) living in naturalistic social groups on a 475-acre South Carolina barrier island. Blood and cerebrospinal fluid (CSF) samples were obtained during random trappings, and the subjects were located for observation by radio telemetry. Quantitative behavioral samples totaling 203 observation hours were taken during two mating seasons (September through January) in 1994 and 1995. Control observations (65 h) on 13 subjects were also taken during the non-mating seasons in 1994 and 1995. The results indicate that CSF 5-hydroxyindoleacetic acid (5-HIAA), CSF testosterone, and plasma testosterone concentrations increase significantly during the mating season. During the mating season, there were significant increases in high intensity aggression, low intensity aggression, grooming behavior, and heterosexual mounting. In the mating season, CSF 5-HIAA was significantly correlated with several sociosexual behaviors: consorts per hour, heterosexual mounts per hour, and inseminations per hour. In contrast to previous findings from the non-mating season, CSF 5-HIAA was not correlated with any measures of aggression or sociality, although during consorting, CSF 5-HIAA was positively correlated with grooming. From these findings, we conclude that the lack of correlation between intense and severe aggression and CSF 5-HIAA in the mating season may reflect the use of high intensity aggression in 'normative' male-male competition over access to reproductively active females. We also conclude that CNS serotonin turnover is positively correlated with sexual competence, i.e. males with low CSF 5-HIAA concentrations are less sexually competent than males with higher concentrations. PMID- 9335202 TI - Hemispheric functional imbalance in a sub-clinical obsessive-compulsive sample assessed by the Continuous Performance Test, Identical Pairs version. AB - Obsessive-compulsive disorder (OCD) sufferers have long been observed to give excessive consideration to normally ignored exogenous and endogenous stimuli. This over-focused attention concerning their symptoms has led researchers to experimentally investigate the attentional mechanisms involved in this disorder and its psychobiological basis. Previous psychometric and neuropsychological research has demonstrated the validity of the sub-clinical analogue in the study of the mechanisms underlying OCD. In this study, 71 introductory university students were recruited from an original pool of 450 people on the basis of their scores on the Spanish version of the Padua Inventory. A high obsessive group (n = 35) was compared with a control group (n = 36) on a standard sustained attention task: the Continuous Performance Test, Identical Pairs version (CPT-IP). The results showed a significant interaction effect between CPT-IP subscales (verbal and spatial) and group membership. This effect was more evident among men. The results were unrelated to general intelligence, depression, anxiety, personality or motivational factors. These findings support the hypothesis that neuropsychological deficits in OCD may be related to a hemispheric functional imbalance rather than to a lateralised dysfunction of a particular hemisphere. PMID- 9335203 TI - Dose-dependent reduced haloperidol/haloperidol ratios: influence of patient related variables. AB - Plasma reduced haloperidol (RH) concentrations or RH to haloperidol (HL) ratios have been suggested to be important in determining the clinical efficacy and extrapyramidal side effects of HL. In this study, we measured the steady-state plasma HL and RH levels by high performance liquid chromatography and analyzed the effects of various variables (dose, gender, age, and body weight) on RH/HL ratios in four dose groups of Chinese schizophrenic inpatients: 10 mg/day (n = 84), 20 (n = 111), 30 (n = 29), and 60 (n = 55). In addition, the polymorphic distribution of RH/HL ratios, suggested by previous investigators, was further tested in each dosage group (for controlling the potential dosage effect on RH/HL ratios). As a result, both age and body weight could influence RH/HL ratios. Each year increase in age (after adjusting the effects of gender, body weight, and dosage) would elevate the RH/HL ratio by 0.0067 (P < 0.0001). On the other hand, after adjusting gender, age, and dosage effects, each kg increment in body weight would decrease the RH/HL ratio by 0.0044 (P < 0.01). Gender did not influence the ratio. Furthermore, the high dosage groups had higher RH/HL ratios (even with other variables being controlled). In comparison with the 10 mg group, the 60 mg group exhibited a higher mean RH/HL ratio by 0.84 (P < 0.0001) and the 30 mg group did by 0.31 (P < 0.0001). The 20 mg group was almost equal to the 10 mg group in RH/HL ratios. Besides, at each dosage group, the frequency distribution of RH/HL ratios seemed to be predominantly unimodal with a small proportion of extreme outliers. The results of this study clearly indicate that aging or a high dose (> or = 30 mg/day) of HL could raise the plasma RH/HL ratio, while an increasing body weight would reduce that. In contrast, gender does not affect the ratios. PMID- 9335204 TI - The effects of beta-adrenoceptor antagonists on a rat model of neuroleptic induced akathisia. AB - Neuroleptic-induced defecation in rats in a well-habituated environment has been proposed as a model of the subjective component of akathisia. In this study, we examined the effects of two lipophilic beta-adrenoceptor antagonists - the non selective drug propranolol and the relatively beta1-selective metoprolol - and one non-selective hydrophilic drug nadolol in this model. Young male Wistar rats were randomly assigned to one of eight groups (n = 12 in each group) and treated with haloperidol or vehicle, with or without one of the beta-antagonists. Haloperidol-treated rats had higher bolus counts than vehicle-treated rats, and this increase was significantly reversed by the lipophilic but not the hydrophilic beta-antagonists. This finding is consistent with the reported anti akathisia effects of these drugs in humans, suggesting that this effect is central in origin and achievable with relatively selective beta1-antagonism. The B-antagonist drugs significantly reduced the cataleptic effect of haloperidol and this effect warrants further examination. PMID- 9335205 TI - A preliminary study on early onset schizophrenia and bipolar disorder: large polyglutamine expansions are not involved. AB - Genetic factors are of major aetiological importance in bipolar disorder and schizophrenia. The exact mode of inheritance is unknown, but recent arguments in favor of genetic anticipation in those two disorders suggest that dynamic mutations could be involved. Using a new antibody, we thus explored the implication of large expanded polyglutamine tracts in a sample of very early onset schizophrenic and bipolar patients. No evidence for a specific protein with polyglutamine expansion was found in either group. PMID- 9335206 TI - An open study of lamotrigine in refractory bipolar depression. AB - Bipolar depressed patients (n = 22) who were refractory to treatment with a combination of divalproex sodium (DVP) and another mood stabilizer or DVP and an antidepressant for 6 weeks were treated in an open naturalistic study with an addition of lamotrigine to DVP. Sixteen out of 22 (72%) responded by the end of week 4 and none developed rash or switched to mania. The results of this preliminary study suggest that lamotrigine may be useful in bipolar depression. PMID- 9335223 TI - Possible association of catecholamine turnover with the polymorphic (TCAT)n repeat in the first intron of the human tyrosine hydroxylase gene. AB - Five allelic fragments were typed by a PCR-based process with a pair of primers specific for the polymorphic sequence due to (TCAT)n tetranucleotide repeat, a microsatellite repeat, in the first intron of the human tyrosine hydroxylase gene, i.e. A1, A2, A3, A4 and A5. Comparisons of some neurochemical parameters of the catecholamine pathway were then made between the unrelated individuals genotypically classified into six subgroups, five homozygotic and one heterozygotic. Among the six subgroups, the individuals with the A2/A2 genotype had the highest levels of serum noradrenaline and those with the A4/A4 genotype had the lowest, and the individuals with the A1/A1 genotype had the highest levels of serum homovanillic acid. These findings suggest that polymorphism of the (TCAT)n repeat in the first intron of the human tyrosine hydroxylase gene may be associated with catecholamine turnover. Possibly, the two alleles, A2 and A4, may be related to the high and low excitabilities of the noradrenergic nerves, respectively, and the allele, A1, may be associated with the up-regulation of dopamine turnover. PMID- 9335222 TI - Involvement of protein kinase C in the supersensitivity to 5-HT caused by oxidized low-density lipoproteins. AB - The effect of native (n-LDL) and oxidized (ox-LDL) low-density lipoproteins and lysophosphatidylcholines (LPCs) on: (1) vasodilator responses induced by acetylcholine (ACh) in intact rabbit aorta segments, and (2) vasoconstrictor responses to serotonin (5-HT), and potassium (K+) in endothelium denuded segments was investigated. In intact vessels, 100 microg/ml ox-LDL did not modify ACh induced relaxation, while it was diminished by 300 microg/ml ox-LDL and abolished by 50 microM LPCs. In contrast, this relaxation was unaltered by n-LDL (100 or 300 microg/ml). In deendothelialized arteries, 100 and 300 microg/ml n-LDL as well as 50 microM LPCs did not modify the contractions induced by 5-HT or K+, while 100 or 300 microg/ml ox-LDL increased the 5-HT-induced contraction, without altering those induced by 75 mM K+. Incubation with 100 or 300 microg/ml ox-LDL increased the contractile response to the protein kinase C (PKC) activator phorbol 12,13-dibutyrate (PDB) (0.1-1 microM) in a concentration-dependent manner, which was blocked by staurosporine (0.1 microM), and unaltered by (50 microM) calphostin C or (50 microM) chelerythrine, the three are PKC inhibitors. Preincubation with 0.05 microM PDB increased the contraction elicited by 5-HT, while staurosporine decreased the PDB-induced contraction, and prevented the 5-HT response increase caused by 300 microg/ml ox-LDL. These results suggest that only ox-LDL reduces endothelium-dependent relaxation and elicits PKC activation, and that this activation mediates, at least in part, the vasoconstrictor response to 5-HT. PMID- 9335224 TI - Characterization and stimuli for production of pericardial fluid atrial natriuretic peptide in dogs. AB - Recently high immunoreactive atrial natriuretic peptide (ir-ANP) levels have been found in the pericardial fluid of patients undergoing cardiac surgery. The present study was designed to characterize pericardial fluid ANP in anesthetized dogs. Pericardial fluid ir-ANP levels were 3.4-fold higher than plasma levels and the molecular form, revealed by high performance liquid chromatography, was indistinguishable from ANP[99-126]. Elimination of [125I]ANP was 5-fold slower in the pericardial space than in plasma. Activity of the major ANP degrading enzyme, neutral endopeptidase (NEP, EC 3.4.24.11), was 15-times higher in the pericardial fluid than in plasma. Right atrial balloon distension and rapid right ventricular pacing induced maximally 2.3-fold and 1.5-fold increases of pericardial fluid ir ANP, respectively. Pericardial fluid ir-ANP concentrations and right atrial pressure values showed significant correlation during the stimuli. Our present results show that high concentrations of ir-ANP can be found in the dog pericardial fluid even under unstimulated conditions. Slow elimination of ANP from the pericardial fluid compartment may contribute to the high peptide levels. However this slow elimination cannot be attributed to a lower NEP activity. High basal levels of ANP in the pericardial fluid could be further increased by atrial balloon stretch and rapid ventricular pacing. The increase of pericardial fluid ir-ANP appeared to be a stretch-dependent response. ANP released into the pericardial fluid may be involved in the regulation of cardiac function and coronary vascular tone. PMID- 9335225 TI - Species differences in the effects of an endogenous inotropic factor (EIF) on the myocardium and aortic smooth muscle. AB - Possible species differences and organ specificity in contractile or relaxation effects of an endogenous inotropic factor (EIF) isolated and purified from porcine heart left ventricle were examined in guinea pig and rat isolated atria, trabeculae and aortic smooth muscle rings. EIF demonstrated a dose-dependent increase in contractile force in guinea pig and rat atria and right ventricle trabeculae. The magnitude of the contractile effect was significantly lower in both the preparations of rat species as compared with guinea pig. In rat isolated aortic rings, EIF had no effect on the basal muscle tone. However, when rings were pre-contracted with phenylephrine EIF caused dose-dependent relaxation of the muscle. When aortic rings isolated from guinea pig were used, EIF induced a dose-dependent contraction which could be blocked by pre-treating the rings with either 2 microM phentolamine or 20 microM prazosin. When these same pre-treated rings were pre-contracted with histamine before the addition of EIF, a dose dependent relaxation of guinea pig aortic smooth muscle rings was observed. Also depletion of catecholamines from the pre-synaptic nerve terminals innervating the aortic rings, using either 0.6 mM rauwolscine or reserpinizing (5 mg/Kg) the guinea pig 24 hours before sacrificing the animal, completely prevented EIF induced contraction of the aortic rings. Instead EIF caused a dose dependent relaxation of the histamine pre-contracted aortic rings similar to that observed in rat aortic rings. The relaxation effect of EIF was demonstrated to be endothelium dependent and nitric oxide mediated in both species since EIF-induced relaxation could be inhibited by 2 microM L-NAME, a nitric oxide synthase inhibitor. Atropine (0.2 microM) or Indomethacin (10 microM) had no significant effect on EIF-induced relaxation of either guinea pig or rat aortic smooth muscle. PMID- 9335226 TI - Thromboxane A2 synthetase inhibitor failed to ameliorate the arterial narrowing during the chronic phase of cerebral vasospasm. AB - To determine the pathogenetic mechanism underlying the maintenance of arterial narrowing during the chronic phase of cerebral vasospasm caused by subarachnoid hemorrhage (SAH), we examined the effect of ozagrel, a thromboxane A2 (TXA2) synthetase inhibitor, on chronic vasospasm in a canine two-hemorrhage model in comparison with that of fasudil, an inhibitor of protein kinases. The magnitude of the vasospasm was determined angiographically. On SAH day 7, a vasospasm was observed in every dog. Intraarterial or intravenous administration of ozagrel (3 mg/kg/30 min) did not reverse the vasospasm but tended to increase bleeding. In contrast, intraarterial administration of fasudil (3 mg/kg/30 min) significantly reversed the vasospasm. These findings suggest that: 1) TXA2 does not participate in the maintenance of chronic vasospasm after SAH; and 2) the protein kinases, particularly myosin-light chain kinase and protein kinase C, are involved in the pathogenesis of arterial narrowing during the chronic phase of cerebral vasospasm. PMID- 9335227 TI - The beta adrenoreceptor antagonist, nipradilol, preserves the endothelial nitric oxide response in atherosclerotic vessels of rabbit. AB - We evaluated the effects of nipradilol, a beta-adrenoreceptor antagonist which contains a nitroxy residue, for vascular response in atherosclerosis of rabbits. Four groups of rabbits received different diets (standard diet; standard diet plus 10 mg/kg/day nipradilol; atherogenic diet [standard diet plus 1% cholesterol]; atherogenic diet plus 10 mg/kg/day nipradilol) for 9 weeks. Plasma lipids, blood pressure, vascular function, nitric oxide (NO), activity of NO synthase, cGMP, and histological atherosclerotic changes were evaluated. Neither the atherogenic diet nor nipradilol treatment affected significantly the animals' body weight, blood pressure, or heart rate. The atherogenic diet increased total cholesterol and triglycerides, which were not altered by nipradilol. The atherogenic diet diminished the acetylcholine-induced NO mediated relaxation. Nipradilol treatment restored this relaxation. Analyses using a NO-sensitive selective electrode showed that nipradilol released NO in the presence of cells and that NO release was greater in atherosclerotic aorta with than without nipradilol treatment. Nipradilol treatment increased the basal NO release as evaluated by the aortic tissue cyclic GMP (cGMP) levels in atherosclerotic vessel, and reduced the esterified cholesterol levels in atherosclerotic vessel. Conclusively, NO released by nipradilol may protect endothelium derived relaxation in atherosclerotic vessels, and may partially inhibit the accumulation of cholesterol in the atherosclerotic lesions. PMID- 9335228 TI - Specific expression of ES46.5K, a novel microsomal esterase, in the mouse liver and its catalytic activity for xenobiotic amide and esters. AB - ES46.5K, a novel esterase, was found in mouse hepatic microsomes. The enzyme catalyzed hydrolysis of 2-acetylaminofluorene and cannabinoid esters. In latter case, the enzyme regioselectively hydrolyzed acetates of 11-hydroxy-delta8 tetrahydrocannabinol. Western immunoblotting analysis demonstrated that none of immuno-reactive proteins against ES46.5K were present in hepatic microsomes from rats, guinea-pigs, monkeys and humans. Rabbit hepatic microsomes contained an immuno-reactive protein, although molecular weight of the protein was rather high (50 kDa) by SDS-PAGE. Esterase activity stained after PAGE demonstrated that ES46.5K retained at origin. Hepatic microsomes of above animal species contained several activity bands on the PAGE, while only mouse hepatic microsomes exhibited significant activity at origin. In mice, liver was only an organ containing ES46.5K by analyzing Western immunoblotting. These results indicate that distribution of ES46.5K is quite different from known carboxylesterases, and suggest that the enzyme has some role in the biotransformation of xenobiotic amide and esters. PMID- 9335230 TI - A PCR-based non-radioactive X-chromosome inactivation assay for genetic counseling in X-linked primary immunodeficiencies. AB - The Wiskott-Aldrich syndrome (WAS), X-linked severe combined immunodeficiency (SCIDX1), and X-linked agammaglobulinemia (XLA) are severe congenital immunodeficiencies with X-linked inheritance. Although rare, they are all associated with severe infections from early in life, and high morbidity and mortality. Female carriers of these diseases can be identified by a non-random pattern of X-chromosomal inactivation in cell lineages targeted by each gene defect. For patients with WAS, SCIDX1 or XLA, the demonstration of non random X Chromosome inactivation in their mothers can be used to confirm clinical diagnosis. Furthermore, analysis of X-Chromosome inactivation in at risk females allows preconceptional carrier detection, thus representing an important aid in genetic counseling. For each disease we established a PCR-based, non radioactive assay at the human androgen receptor (HUMARA) locus, that allows analysis of X Chromosome inactivation in the affected cell types and in tissue specific controls to exclude the issue of skewed X-chromosomal inactivation. In our study, 50 females with a known family history of XLA [19], WAS [18], and SCIDX1 [13],were examined. A carrier status was established in 19 females (7 XLA, 6 WAS, 6 SCIDX1) and excluded in 29 ( 11 XLA, 11 WAS, 7 SCIDX1). Only in 2 cases (4%) the assay was not informative. PMID- 9335229 TI - Prolactin modulation of nitric oxide and TNF-alpha production by peripheral neutrophils in rats. AB - It has been demonstrated that prolactin (PRL) is a potent immunomodulator that exerts stimulatory effects on physiological responses of immune cells. In the present research we have investigated whether PRL may influence nitric oxide (NO) and/or tumor necrosis factor-alpha (TNF-alpha) production in neutrophils obtained from inflammatory exudate of carrageenin-induced experimental pleurisy in the rat. In this acute model of inflammation the role of endogenous NO was evaluated using an inhibitor of NO-synthase, NG-nitro-L-arginine methyl ester (L-NAME). A treatment of animals with L-NAME (10 mg/kg s.c.) induced a reduction of volume and cell number of pleural exudate and a decrease of nitrite production (measured by the Griees reaction) by polymorphonuclear cells after 24 h of incubation, while D-NAME, the inactive isomer, was without effect. Neutrophils from ovine prolactin (oPRL) treated rats (5 mg/kg for 5 times s.c.) or from rats with a hyperprolactinaemia induced by pituitary gland graft produced higher amounts of NO both after 24 and 48 h of incubation. On the contrary, a clear reduction in the production of NO was found in neutrophils from rats treated with bromocriptine (BRC) (2 mg/kg s.c.), a dopamine D2-receptor agonist. TNF-alpha production (measured by MTT/cytotoxic assay) by neutrophils was markedly increased in PRL-treated or pituitary-grafted rats in comparison to controls, whereas BRC treatment reduced TNF-alpha production. PMID- 9335231 TI - Allixin, a phytoalexin produced by garlic, and its analogues as novel exogenous substances with neurotrophic activity. AB - Effects of allixin, a phytoalexin of garlic, and its analogues were studied on the survival and morphology of primary cultured neurons from fetal rat brain. Addition of allixin (1-100 ng/ml) to medium significantly promoted the survival of neurons derived from various regions of brain and increased the number of branching points per axon in hippocampal neurons. Allixin, however, was cytotoxic at higher concentrations (>1 microg/ml). Among the analogues of allixin, 2,6 dimethyl-3-hydroxy-4H-pyran-4-one (DHP) possessed potent neurotrophic activity at concentrations over 10 ng/ml without any obvious cytotoxicity up to 10 microg/ml. DHP also retained the activity to promote axonal branching. These results indicate that DHP is a novel exogenous low molecular weight neurotrophic substance without apparent cytotoxicity. This compound may be a useful prototype leading chemical for developing therapeutic and/or prophylactic drugs for neurodegenerative disorders. PMID- 9335232 TI - Combat veterans with posttraumatic stress disorder exhibit decreased habituation of the P1 midlatency auditory evoked potential. AB - The current study used a paired stimulus paradigm to investigate the P1 midlatency auditory evoked potential in Vietnam combat veterans with posttraumatic stress disorder (PTSD) and three comparison groups: alcohol dependents, combat-exposed normals, and combat-unexposed normals. Compared to each comparison group, PTSD subjects exhibited significantly diminished habituation of the P1 potential. P1 potential habituation within the PTSD group, correlated significantly with intensity of PTSD reexperiencing symptoms, such as trauma-related nightmares and flashbacks. These findings are discussed as consistent with a sensory gating defect at the brainstem level in PTSD, and are further discussed in the context of other psychophysiological measures in PTSD and of P1 potential findings in psychiatric disorders other than PTSD. PMID- 9335233 TI - Effect of phorbol ester on the regulation of LH/hCG receptors. AB - Granulosa cells have been used to study the regulation of LH/hCG receptor protein and mRNA expression. Phorbol 12-myristate 13-acetate (PMA) dose-dependently attenuates the increases in LH/hCG receptor mRNA and protein induced by FSH and forskolin (FSK). The presence of PMA caused a decrease in cAMP production stimulated by FSH and FSK. These results suggest that PMA-mediated decreases in cAMP are a major factor in PMA-mediated decreases in LH/hCG receptor mRNA. On the other hand, in the presence of 8-Br-cAMP, PMA significantly increased LH/hCG receptor mRNA and protein, with maximal stimulation between PMA concentrations of 3 to 30 nM (1.5 fold) with 8-Br-cAMP. These findings suggest that activation of protein kinase C by PMA attenuates the increase in cAMP accumulation induced by FSH but enhances the effect of cAMP on LH/hCG receptor expression, and that the inhibitory and stimulatory effects of PMA on LH/hCG receptor content are correlated with regulation of LH/hCG receptor mRNA levels. Since the half-life study revealed no change in the stability of the LH/hCG receptor mRNA following PMA treatment, a change in the rate of LH/hCG receptor gene transcription must be responsible for the change in the LH/hCG receptor mRNA levels. PMID- 9335235 TI - How subtle is too subtle? The histopathologic diagnosis of "early" malignancies of the skin. PMID- 9335234 TI - Binding of the non-classical cannabinoid CP 55,940, and the diarylpyrazole AM251 to rodent brain cannabinoid receptors. AB - The binding of [123I]AM251 (a radioiodinated analog of the cannabinoid CB1 receptor antagonist SR141716A) was compared to that of [3H]CP 55,940 in mouse and rat brain preparations. Scatchard analysis of the binding of [123I]AM251 and [3H]CP 55,940 to membranes prepared from mouse cerebellum, striatum and hippocampus yielded similar Bmax values (15-41 pmol/g wet wt tissue). Kd values were lower for [123I]AM251 (0.23-0.62 nM) than for [3H]CP 55,940 (1.3-4 nM). CP 55,940 and SR141716A increased dissociation of [123I]AM251 from binding sites in mouse cerebellar homogenates to a similar extent. The structurally dissimilar cannabinoid receptor ligands THC, methanandamide, WIN 55, 212-2, CP 55,940 and SR141716A were each able to fully compete with binding of both [123I]AM251 and [3H]CP 55,940 in mouse cerebellum. In vitro autoradiography demonstrated that the distribution of binding sites for [123I]AM251 in rat brain was very similar to published distributions of binding sites for [3H]CP 55,940. Together, these observations suggest that AM251 binds to the same site (the cannabinoid CB1 receptor) in rodent brains as CP 55,940. However, the binding site domains which interact with AM251 and CP 55,940 may not be identical, since IC50 values for cannabinoid receptor ligands depended on whether [123I]AM251 or [3H]CP 55,940 was used as radioligand. PMID- 9335236 TI - Solar keratosis can be distinguished from superficial basal cell carcinoma by expression of bcl-2. AB - Criteria for the histological diagnosis of solar keratosis (SK) and superficial basal cell carcinoma (SBCC) are well defined. On occasion, however, the distinction can be difficult, particularly when evaluating small-punch or superficial shave biopsies in which the overall global architectural pattern of the lesion is not represented. As bcl-2 expression has been established in basal cell carcinoma, this study was undertaken to determine whether it could be employed as a basis for distinguishing SBCC from SK. Ten cases each of typical SBCC and SK were studied. Every example of SBCC showed strong uniform expression of the bcl-2 gene product Bcl-2, whereas all examples of SK were uniformly negative. It is suggested, therefore, that this simple technique is a reliable method for distinguishing these two closely related but disparate lesions. PMID- 9335237 TI - Atypical lymphoid infiltrates arising in cutaneous lesions of connective tissue disease. AB - Atypical lymphoid infiltrates occurring in the setting of connective-tissue disease (CTD) comprise malignant neoplasms of B-cell or T-cell phenotypes and various reactive lymphoid hyperplasias, such as myoepithelial sialadenitis, lymphocytic thyroiditis, and lymphocytic interstitial pneumonitis. We describe 17 patients with atypical lymphoid infiltrates arising in cutaneous lesions of CTD, the spectrum of which included lupus erythematosus, dermatomyositis, relapsing polychondritis, and lichen sclerosus et atrophicus. There were two principal categories, pseudolymphoma and malignant lymphoma, the former representing 15 of the 17 cases. The clinical and histologic features and possible pathogenetic mechanisms are discussed. PMID- 9335239 TI - Anatomic distribution of melanocytes in normal nail unit: an immunohistochemical investigation. AB - Very few histologic reports describe normal melanocytes of the nail unit. Previous studies predominantly address the distal nail matrix melanocytes; we found no review of nail-bed melanocytes in the literature. The proximal nail matrix melanocytes are difficult to identify; the cells cannot be identified by L DOPA staining. More recently, their scarcity was confirmed by immunohistochemistry with a large panel of antibodies directed against melanocytes. We wished to detect the proximal nail matrix dormant melanocytes and compare their density and distribution with that of the other melanocytes in the distal matrix and nail bed and to establish criteria of normality that may help clarify the pathologic features of benign nevoid melanonychia in the nails of whites. A panel of five monoclonal antibodies (MoAbs), including HMB45 and TRP1 directed against antigens localized in early melanosomal vesicles, was investigated in frozen sections of six nail specimens from whites. Both vertical and horizontal sections were assessed to determine the presence of dormant melanocytes. Results showed that the proximal nail matrix melanocytes were clearly identified with MoAbs HMB45 and tyrosinase-related protein-1 (TRP-1). By contrast, melanocytes stained by MoAb against tyrosinase and L-DOPA reaction were evident, especially in the distal matrix. With MoAb TRP-1, the epithelial sheets showed counts of approximately 217+/-84/mm2 in the proximal matrix and of 132+/ 34/mm2 in the distal matrix; the nail bed counts were only 45+/-25/mm2. The split epithelial sheets had 103+/-17/mm2 L-DOPA-positive melanocytes in the distal third of the matrix, but only a few of them were detected in the proximal matrix and none were noted in the nail bed. We clearly identified proximal nail melanocytes using MoAb HMB45 and TRP1. The total number of matrix melanocytes can be estimated as approximately 217/mm2. In proximal matrix, the dormant melanocytes compartment was predominant. In the distal matrix, two compartments were identified: a functionally differentiated and a dormant compartment. Contrary to classical opinion, longitudinal melanonychia originates more frequently in the distal matrix, not secondary to the larger melanocyte density but because only the distal matrix contains an active melanin synthesis compartment. Furthermore, the superficial distribution of proximal nail melanocytes in vertical sections showed a histologic feature that may simulate the pagetoid pattern of melanoma in situ. PMID- 9335238 TI - Histogenesis of mixed tumor of the skin, apocrine type: immunohistochemical study of keratin expression. AB - To investigate the histogenesis of mixed tumor of the skin, apocrine type, the immunophenotypes of 10 cases were examined using 19 different monoclonal anti keratin antibodies and antibodies against carcinoembryonic antigen (CEA) and involucrin. By using light microscopy, four epithelial elements in this tumor were characterized: tubular branching structures with lumina lined by cuboidal epithelium and those with lumina lined by columnar epithelium, keratinous cysts, and solid aggregates of epithelial cells. The immunohistochemical patterns of cytokeratin expression suggested that cuboidal and columnar cells differentiated, respectively, toward the ductal and secretory cells of apocrine glands, whereas keratinous cysts revealed follicular infundibular differentiation. Furthermore, CEA expression, a marker for sweat-gland differentiation, was present not only on tubules' luminal surfaces but also on the inner surfaces of keratinous cysts. The simultaneous coexpression of CEA and cytokeratins specific for follicular infundibulum in the keratinous cysts, although perplexing, suggested that keratinous cysts may contain some cells differentiating toward the intrafollicular portion of apocrine ducts that enter infundibulae rather than eccrine ducts that have no infundibular association. We conclude that apocrine type of mixed tumors of the skin demonstrate differentiation toward all components of apocrine units. PMID- 9335241 TI - Malignant melanoma masquerading as malignant fibrous histiocytoma. AB - Numerous cytological variants of malignant melanoma (MM) exist. We report two cases of MM resembling malignant fibrous histiocytoma (MFH). Pathologists should be aware of this variant. Possible pitfalls that could lead to an incorrect diagnosis are discussed. PMID- 9335240 TI - Silver-stained nucleolar organizer regions in hypertrophic and keloid scars. AB - Silver-stained nucleolar organizer regions (AgNORs) have been widely used as a marker of cellular activity and proliferation. In a retrospective study, we investigated the potential value of AgNORs in 12 hypertrophic and 24 keloid scar tissues. Ten normal skin tissues served as controls. A standard silver-staining method was used, and the mean AgNOR count of dermal fibroblastic cells in each tissue was determined. In normal skin, the mean AgNOR count of dermal fibroblasts was 1.79+/-0.55, whereas fibroblastic cells in hypertrophic and keloid scars had mean AgNOR counts of 3.18+/-0.56 and 5.10+/-0.97, respectively. There was a statistically significant difference between the mean AgNOR counts of fibroblastic cells from normal skin, hypertrophic scar, and keloid scar [one factor analysis of variance (ANOVA), p < 0.0001]. Our findings suggest that AgNOR count may be a useful marker for assessment of fibroblastic cell activity in hypertrophic and keloid scars, which may have potential value for histologic and biologic characterization of the two lesions. PMID- 9335242 TI - Characterization of the cutaneous inflammatory infiltrate in canine atopic dermatitis. AB - Sections from lesional atopic, clinically normal atopic, and normal canine skin were investigated by light microscopy and an immunoperoxidase method using monoclonal antibodies specific for canine leukocyte antigens. We confirmed that skin-infiltrating cells of canine atopic dermatitis are constituted of mast cells, dendritic antigen-presenting cells, memory helper T-lymphocytes, low numbers of eosinophils and neutrophils, and rare B-lymphocytes. The presence of epidermal eosinophil microaggregates and clustered Langerhans' cells supports the hypothesis of epidermal allergen contact. The hyperplasia of epidermal T-cells expressing the gamma/delta T-cell receptor appears specific to canine atopic dermatitis compared with its human counterpart. This finding could be explained by an interspecies difference in skin immune systems or, alternatively, by an active participation of these epitheliotropic gamma/delta T-cells in the cutaneous allergic immune response in dogs. The paucity of dermal neutrophils in spontaneous lesions of canine atopic dermatitis is notably different from the neutrophil-rich late-phase reactions provoked by intradermal allergen injections in allergic dogs. This difference in the cellular infiltrate probably results from variations in the immune reaction between single and repeated allergen exposure as well as epidermal versus dermal antigen contact. PMID- 9335243 TI - The interdependence of dermatopathology and basic science: Joint Meeting of the 17th Colloquium of the International Society of Dermatopathology and the Swiss Group of Dermatopathology, Zurich, Switzerland, July 18-20, 1996. PMID- 9335244 TI - Solitary fibrous tumor of the skin. AB - Solitary fibrous tumor (SFT) is a rare mesenchymal neoplasm that most commonly involves the pleura, but is increasingly more often observed in extrapleural locations. A 37-year-old woman presented with an SFT involving the skin and subcutaneous tissue of the scalp. Histologically, SFT is well circumscribed and composed of uniform spindle cells arranged in interlacing fascicles. It exhibits alternating hypercellular and hypocellular areas with abundant thick, often keloid-like, hyalinized collagen. Hemangiopericytoma-like areas are frequently prominent. Immunohistochemical markers for smooth muscle, neural, and epithelial differentiation are negative, but generalized positivity for CD-34 is characteristic. Because of the expanding spectrum of anatomic involvement of SFT, it is not surprising that on rare occasions this tumor may involve the skin. PMID- 9335245 TI - Cutaneous epithelioid cell histiocytoma: immunohistochemical and ultrastructural findings suggesting endothelial origin. AB - A 13-year-old boy presented with a polypoid nodule localized in the groin. Although the clinical and histopathological features corresponded to previously described diagnostic criteria of epithelioid cell histiocytoma, immunohistochemical and ultrastructural findings suggested vascular differentiation of the epithelioid cells. In particular, the immunohistochemical negativity of the epithelioid cell elements for Factor XIIIa failed to substantiate the previously forwarded hypothesis that this lesion represents a dendrocytoma. Instead, the presence of histiocytoid, vacuolated cells occasionally containing sparse red blood cells, positive staining for Factor VIII related antigen, and ultrastructural evidence of endothelial characteristics in epithelioid neoplastic cells favor the hypothesis that "epithelioid cell histiocytoma" is of endothelial origin. We suggest the descriptive term cutaneous histiocytoid hemangioendothelioma for this lesion. PMID- 9335246 TI - Agminated intradermal Spitz nevi arising on an unusual speckled lentiginous nevus with localized lentiginosis: a continuum? AB - We report an 18-year-old boy with a congenital pigmented lesion measuring 2 x 6 cm on his right thigh. About a third of the lesion was composed of numerous lentiginous macules superimposed on histologically normal and clinically nontan skin; in the remainder of the lesion, several macules and papules with histologic features of junctional and compound nevi were superimposed on clinically normal skin, which had a lentiginous pattern histologically. Some years later, eruptive intradermal Spitz nevi developed at one corner of the lesion. The combined clinical and histological features of the lesion fulfill descriptions for both segmental lentiginosis and an unusual variant of speckled lentiginous nevus. Our case points out the limitations of using strict diagnostic criteria to define speckled lentiginous nevus and offers an opportunity to consider the natural history of the lesion as a continuum from lentigines to melanocytic nevi. Moreover, the presence of eruptive intradermal Spitz nevi arising within the area of speckled lentiginous nevus lacking a distinct tan background, suggests the possibility that the entire area of the lesion per se constitutes an environment where development of nevi is enhanced. PMID- 9335247 TI - Metastatic inflammatory carcinoma of the rectum: tumor spread by three routes. AB - We report a case of a patient with adenocarcinoma of the rectum with inflammatory metastases to the skin who was treated with radiation therapy and subsequently developed lymphatic obstruction with resultant extensive lymphedema of the lower extremities. Histopathologic examination and immunohistochemistry showed intralymphatic, intravascular, and interstitial malignant cells in the dermis. To our knowledge, this is the second reported case of inflammatory carcinoma arising from metastatic carcinoma of the rectum. However, tumor cell spread to the skin by three routes has not previously been demonstrated. PMID- 9335248 TI - Syringoid eccrine carcinoma: a case report. AB - We report a case of syringoid eccrine carcinoma, a rare syringomatous tumor of the skin, occurring in a 70-year-old woman. Histological and immunohistochemical criteria are given to differentiate this neoplasm from other primary carcinomas of the skin as well as from skin metastases of internal malignancies. PMID- 9335249 TI - Malignant proliferating trichilemmal tumor showing distant metastases. AB - We describe a case of malignant proliferating trichilemmal tumor showing multiple distant metastases. For 10 years, the patient had had a round mass in the occiput, which recurred twice after wide excisions, and later metastasized to the cervical lymph nodes, periparotid area, and chest. Each time the lesions were excised, histologic specimens demonstrated a proliferating trichilemmal tumor with increasing nuclear atypia. Serial specimens showed increasing Ki-67 positivity as the extent of the tumor advanced. PMID- 9335250 TI - Clear-cell porocarcinoma: another cutaneous marker of diabetes mellitus. AB - The relationship between clear-cell syringoma and diabetes mellitus is well established. We present a case of clear-cell porocarcinoma in a diabetic patient. The lesion consisted of a 5-cm nodule on the lateral aspect of the left leg. Histopathologically, the neoplasm was composed of irregular aggregations of neoplastic cells with striking clear-cell appearance, showing features of ductal differentiation. The clear-cell appearance of neoplastic cells was due to glycogen accumulation within their cytoplasm. Immunohistochemistry and ultrastructural studies also supported the diagnosis of a neoplasm with sweat ductal differentiation. Enzyme histochemical reactions for phosphorylase immunoreactivity on fresh, unfixed sections of the neoplasm demonstrated that this immunoreactivity was remarkably decreased. Some adnexal neoplasms of the skin mostly composed of clear cells may be cutaneous markers of diabetes mellitus. Phosphorylase activity deficiency in diabetic patients may be responsible for glycogen accumulation in neoplastic cells resulting in clear-cell appearance of these neoplasms. PMID- 9335252 TI - What is black bile? PMID- 9335251 TI - Combined neuroendocrine carcinoma of the skin (Merkel cell tumor) and trichilemmal cyst. AB - We report a case of neuroendocrine (Merkel cell) carcinoma (NC) of the skin, associated with a trichilemmal cyst, showing pagetoid spread into the trichilemmal epithelium. The association of the two lesions may strengthen the hypothesis that NC originates from pluripotent stem cells of adnexal epithelium. PMID- 9335253 TI - Estrogen and progesterone receptors in breast cancer. PMID- 9335254 TI - Thy-1 and AvGp50 signal transduction complex in the avian nervous system: c-Fyn and G alpha i protein association and activation of signalling pathways. AB - We have previously reported the isolation of two distinct populations of detergent resistant membrane complexes (DRMC's) from day-old chick brain (Henke et al.: J Neurosci Res 45:617-630, 1996). We now show that the glycosylphosphatidylinositol-anchored proteins Thy-1 and AvGp50 are associated in a signalling complex with c-Fyn, the heterotrimeric G alpha i subfamily members G alpha i-3, G alpha z, and G alpha o, alpha and beta tubulin, and a number of other phosphoproteins in immunocomplexes isolated from both populations of DRMC's. Activation of this signalling complex via Thy-1 monoclonal antibody incubation with chick forebrain cells, elicited a decrease in total phosphoprotein profile and tyrosine kinase activity present in DRMC fractions isolated from these cells, while AvGp50 and control antibodies had no effect. Down-regulation of the DRMC phosphoprotein profile was accompanied by an increase in the Thy-1-associated signalling complex, suggesting that activation of this complex initiates the decreases seen in overall DRMC kinase activity. This signalling complex provides the basis for GPI-anchored protein-mediated signal transduction within the unique plasma membrane domains represented by DRMC's. PMID- 9335255 TI - Analysis of compound heterozygous mice reveals that the Trembler mutation can behave as a gain-of-function allele. AB - The most common form of Charcot-Marie-Tooth disease, CMT1A, is correlated with a 1.5 megabase duplication on chromosome 17p.11.2 containing the peripheral myelin protein 22 (PMP22) gene. Deletion of the same region is associated with a second inherited neural disorder, the hereditary neuropathy with liability to pressure palsies (HNPP). Moreover, several distinct point mutations within the PMP22 coding region are associated with CMT1A and Dejerine-Sottas Syndrome in humans and the Trembler (Tr) and Trembler-J phenotypes in mice. Heterozygous Tr mutants (Tr/+) display severe hypomyelination of peripheral nerve fibers while heterozygous pmp22 knockout mice (pmp22+/0) are characterized by focal hypermyelination. These findings suggest that the Tr mutation does not generate a pmp22 null allele but rather produces its deleterious effects by either a dominant-negative or gain-of-function mechanism. To address this question in detail, we have compared various combinations of pmp22 alleles including Tr/+, Tr/Tr, Tr/0, pmp22+/0, and pmp22(0/0) mice with respect to the resulting myelin abnormalities. The combined analysis of these mutants demonstrates that the Tr allele can act as a true gain-of-function mutation in both, the heterozygous state on a null background (Tr/0) as well as in homozygous Tr animals (Tr/Tr). Furthermore, increasing the relative Tr gene dosage correlates with more pronounced myelin deficiencies and decreased levels of MBP and P0 in 18-day-old mice. PMID- 9335256 TI - Activation of NF-kappaB protects hippocampal neurons against oxidative stress induced apoptosis: evidence for induction of manganese superoxide dismutase and suppression of peroxynitrite production and protein tyrosine nitration. AB - The transcription factor NF-kappaB is expressed in neurons wherein it is activated in response to a variety of stress- and injury-related stimuli including exposure to cytokines such as tumor necrosis factor-alpha (TNFalpha), and excitotoxic and oxidative insults. NF-kappaB may play a role in the anti death actions of TNFalpha in cultured hippocampal neurons exposed to metabolic and oxidative insults. We now report that pretreatment of hippocampal cell cultures with agents that activate NF-kappaB (TNFalpha and C2-ceramide) confers resistance of neurons to apoptosis induced by the oxidative insults FeSO4 and amyloid beta-peptide (Abeta25-35). The neuroprotective actions of TNFalpha and ceramide were abolished in cultures cotreated with kappaB decoy DNA demonstrating a requirement for NF-kappaB activation for prevention of cell death. Levels of manganese superoxide dismutase (Mn-SOD) in neurons were increased following exposure of cultures to TNFalpha and ceramide in control cultures, but not in cultures cotreated with kappaB decoy DNA. FeSO4 and Abeta25-35 induced accumulation of mitochondrial peroxynitrite, and membrane lipid peroxidation, in neurons. Peroxynitrite accumulation and lipid peroxidation were largely prevented in neurons pretreated with TNFalpha and ceramide prior to exposure to FeSO4 and Abeta25-35, an effect blocked by kappaB decoy DNA. Immunoreactivity of neurons with an anti-nitrotyrosine antibody was increased following exposure to FeSO4 and Abeta25-35; TNFalpha and C2-ceramide suppressed protein tyrosine nitration, and kappaB decoy DNA blocked the effects of TNFalpha and C2-ceramide. Finally, the peroxynitrite scavenger uric acid protected neurons against apoptosis induced by FeSO4 and Abeta, and suppressed peroxynitrite accumulation. We conclude that, by inducing production of Mn-SOD and suppressing peroxynitrite formation and membrane lipid peroxidation, NF-kappaB plays an anti-apoptotic role in neurodegenerative conditions that involve oxidative stress. The data further suggest important roles for peroxynitrite and NF-kappaB in the pathogenesis of neuronal degeneration in Alzheimer's disease. PMID- 9335257 TI - Signaling events following the interaction of the neuronal adhesion molecule F3 with the N-terminal domain of tenascin-R. AB - Interaction between the extracellular matrix protein tenascin-R and the neuronal adhesion molecule F3 might be involved in the formation of neuronal networks. In this study, the fragment of tenascin-R comprising epithelial growth factor (EGF) like repeats and the cysteine-rich NH2 terminal stretch (EGF-L), known to be inhibitory for growing neurites and repellent for growth cones, was used to investigate the signaling events following the F3/EGF-L interaction. We addressed this question using an in vitro test with F3-transfected Chinese hamster ovary (CHO) cells that allowed us to measure the kinetics, magnitude and specificity of the repellent effect resulting from the specific F3/EGF-L interaction. We showed that the repellent effect was counteracted by addition of the serine/threonine kinase and -phosphatase modulators (staurosporine, okadaic acid and H7) but not by modulators of tyrosine kinase or -phosphatases. This result indicates that the intracellular signals activated by the repellent effect involve a serine/threonine kinase pathway. Furthermore, the repellent effect of the EGF-L fragment for growth cones of cultured cerebellar neurons was also abolished by the identical modulators of serine/threonine kinase and -phosphatases. The inhibition of neurite outgrowth from hippocampal neurons by EGF-L was abolished in the presence of the serine threonine-kinase inhibitor H7. These results strongly suggest that the F3/tenascin-R interaction through EGF-L involves an intracellular activation of serine/ threonine kinase(s) in all F3-expressing cells tested. PMID- 9335258 TI - Pigment epithelium-derived factor (PEDF) has direct effects on the metabolism and proliferation of microglia and indirect effects on astrocytes. AB - Pigment epithelium-derived factor (PEDF), a neurotrophic agent first identified in conditioned medium from cultured human retinal pigment epithelial cells, induces neuronal differentiation with neurite outgrowth in Y-79 retinoblastoma cells and has a neurotrophic survival effect on cerebellar granule cells in culture. In the present study, we investigated the effects of human recombinant PEDF (rPEDF) on proliferation and activation of microglia and astrocytes isolated from newborn rat brain. rPEDF treatment caused microglia to round up morphologically, increased their metabolic activity (measured by both MTS conversion and acid phosphatase activity), but blocked proliferation (mitosis). This blocking effect could be demonstrated in cultures stimulated to proliferate by addition of granulocyte-macrophage colony stimulating factor. The effect of rPEDF on microglial metabolic activity showed a dose-response relationship both in serum-containing medium and in chemically defined medium and was blocked with anti-PEDF antibody. rPEDF had no direct effect on the metabolic activity or proliferation of cultured astrocytes but blocked their proliferation in astrocyte microglia co-cultures. Proliferation of isolated astrocytes was also blocked by conditioned medium from microglia treated with PEDF (PMCM). The effect of PMCM on astrocytes was not blocked by an antibody to transforming growth factor-beta. These results demonstrate that PEDF activates microglial metabolism while blocking proliferation and suggest that a soluble factor(s) released by rPEDF stimulated microglia blocks the proliferation of astrocytes. Thus, PEDF could play an important role in regulation of glial function and proliferation in the central nervous system. PMID- 9335260 TI - Gp120 can revert antagonism at the glycine site of NMDA receptors mediating GABA release from cultured hippocampal neurons. AB - The effects of the human immunodeficiency virus type 1 envelope protein gp120 on the release of GABA elicited by N-methyl-D-aspartate (NMDA) from rat hippocampal neurons in primary culture has been investigated. NMDA (1-300 microM) increased in a concentration-dependent manner (EC50 =37.9+/-12 microM) the release of [3H] GABA. The effect of 100 microM NMDA was prevented by 30 microM of the GABA transport inhibitor N-(4,4-diphenyl-3-butenyl)guvacine (SKF 100330A). Glycine (10 microM) or gp120 (0.01 microM) affected neither the basal nor the NMDA-evoked [3H]-GABA release. The NMDA (100 microM)-evoked release was prevented by 5,7 dichloro-kynurenic acid (5,7-DCKA), a selective antagonist at the glycine site of the NMDA receptor, in a concentration-dependent manner (IC50 approximately 0.3 microM). Glycine (3-10 microM) or gp120 (0.003-0.01 microM) produced reversal of the 5,7-DCKA antagonism in a way that suggested competition at a same site; gp120 was at least 3 orders of magnitude more potent than glycine. It is suggested that gp120 may mimic glycine at NMDA receptors. PMID- 9335259 TI - Alzheimer's disease-associated presenilin 1 in neuronal cells: evidence for localization to the endoplasmic reticulum-Golgi intermediate compartment. AB - The recently identified Alzheimer's disease-associated presenilin 1 and 2 (PS1 and PS2) genes encode two homologous multi membrane-spanning proteins. Rabbit antibodies to the N-terminal domain of PS1 detected PS1 in human neuroblastoma SH SY5Y wild type and PS1 transfectants (SY5Y-PS1) as well as in mouse P19, in CHO K1 and CHO-APP770 transfected cells, in rat cerebellar granule and hippocampal neurons, and astrocytes. Immunoblotting detected full-length protein of 50 kDa, and a major presumptive cleavage product of 30 kDa. The immunofluorescence pattern resembled labeling of the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) marker protein ERGIC-53. PS1 distribution showed slight condensation after brefeldin A and more marked condensation after incubation of cells at 16 degrees C, characteristic of the ERGIC compartment. Double labeling showed colocalization of ERGIC-53 with PS1 in the SY5Y-PS1 cells. PS1 labeling of SY5Y-PS1 and P19 cells showed overlap of the cis-Golgi marker p210 and colocalization with p210 after brefeldin A which causes redistribution of p210 to the ERGIC. Expression of PS1 did not change in level or cellular distribution during development of neurons in culture. Double labeling for the amyloid precursor protein (APP) and PS1 on SY5Y-PS1 cells and CHO-APP770 cells showed some overlap under control conditions. These results indicate that PS1 is a resident protein of the ERGIC and could be involved in trafficking of proteins, including APP, between the ER and Golgi compartments. PMID- 9335261 TI - Differential effects of basic fibroblast growth factor, epidermal growth factor, transforming growth factor-alpha, and insulin-like growth factor-I on a hypothalamic gonadotropin-releasing hormone neuronal cell line. AB - Recent studies in several neuronal lineages suggest that extrinsic factors such as polypeptide growth factors regulate various stages of neuronal development, from initial commitment of multipotent progenitors to induction of specific gene expression that is characteristic of terminal neuronal differentiation. In the present study, immortalized hypothalamic neurons of the GT1-1 lineage were used to analyze proliferative, as well as morphological and molecular differentiation actions of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), and insulin-like growth factor-I (IGF-I). These effects were compared with those induced by specific activators of protein kinase A and C pathways, which potently inhibited cell proliferation and gonadotropin-releasing hormone (GnRH) gene expression, but stimulated morphological neuronal maturation as determined by the length and number of neurite outgrowth. bFGF exerted a broad spectrum of stimulatory effects, increasing the rate of proliferation measured both by the incorporation of 3H thymidine and by cell number, and parameters of terminal differentiation, such as neurite outgrowth and induction of gene expression. bFGF stimulated the expression of the hybrid transgene-containing portions of the rat GnRH promoter. In contrast, EGF, TGF-alpha, and IGF-I inhibited cell proliferation, while having subtle effects on neurite outgrowth. Thus, GT1-1 cells appear to be differentially responsive to distinct neurotrophic factors, providing a model for studying the specific effects of neurotrophic factors on functional differentiation, migration, and connectivity of hypothalamic neurons. PMID- 9335262 TI - Tryptophan hydroxylase mRNAs analysis by RT-PCR: preliminary report on the effect of noradrenaline in the neonatal rat pineal gland. AB - The levels of mRNAs coding for tryptophan hydroxylase (TPOH), the first enzyme in melatonin synthesis, have been investigated by quantitative reverse transcription of RNA, followed by polymerase chain reaction (RT-PCR), after stimulation of neonatal pineal organ cultures with Noradrenaline (NA). TPOH mRNAs were specifically amplified from various adult tissues, namely the pineal gland, raphe, retina, and kidney, but not the lung. PCR signals for TPOH were detected in the neonatal pineal gland in the absence of stimulation. Stimulation of neonatal pineal organ culture with 0.1 microM NA resulted in a significant increase (x2.5) in expression of TPOH mRNAs, whereas higher doses (1 and 10 microM) had no effect. All concentrations of NA enhanced melatonin secretion. Our results suggest that the level of TPOH mRNAs can be controlled by NA and that this effect might be implicated in the gene level regulation of the daily enzyme rhythm in the rat pineal gland. PMID- 9335263 TI - Beta-amyloid and ionophore A23187 evoke tau hyperphosphorylation by distinct intracellular pathways: differential involvement of the calpain/protein kinase C system. AB - SH-SY-5Y human neuroblastoma cells were treated with 22 microM of a synthetic peptide corresponding to amino acid residues 25-35 of beta-amyloid (betaA) or 3 microM calcium ionophore A23187 in culture medium containing 1.8 mM extracellular calcium. Both agents increased tau immunoreactivity towards antibodies (PHF-1, ALZ-50) that recognize epitopes common with paired helical filaments (PHFs) and towards an antibody (5E2) that recognized a phosphate-independent tau epitope. However, only ionophore increased immunoreactivity with an additional phosphate dependent antibody (AT-8) that recognized an epitope of tau when phosphorylated, and induced a corresponding decrease in immunoreactivity towards an additional antibody (Tau-1) that recognizes the same site when that site is not phosphorylated. Moreover, the ionophore-mediated increase in PHF-1 was blocked by EGTA, by the calpain inhibitor calpeptin and by the PKC inhibitor H7, while that evoked by betaA treatment was not inhibited by any of these treatments. Since ionophore-mediated calpain activation induces proteolytic PKC activation, we further examined the influence of PKC inhibition on betaA and ionophore-mediated PHF-1 induction. Antisense oligonucleotide-mediated downregulation of PKCepsilon in a stable transfectant SH-SY-5Y subclone diminished the ionophore-mediated, but not the betaA-mediated, increase in PHF-1 immunoreactivity. These data indicate specific differences in the intracellular cascade of events invoked by betaA and ionophore A23187. Moreover, although betaA invoked calcium influx in these cells, our findings further suggest that the induction of tau hyperphosphorylation by betaA may not be due to calcium influx. PMID- 9335264 TI - Isolation and culture of bovine endothelial cells of endoneurial origin. AB - Penetration of immunoglobulins and/or migration of activated lymphocytes into peripheral nervous system (PNS) parenchyma are the initial key steps to develop immunological disorders of PNS including Guillain-Barre syndrome, IgM neuropathy and chronic inflammatory demyelinating polyradiculoneuropathy. Hence, it is important to know the cellular property of endothelial cells of endoneurial tissue origin (PnMEC) because these cells constitute the bulk of the blood-nerve barrier (BNB). For this purpose, we developed a method to isolate and culture pure populations of PnMECs from bovine cauda equina. PnMECs were identified by their cobblestone appearance, immunoreactivity against Factor VIII/von Willebrand factor (vWF) antigen, and positive uptake of DiI-Ac-LDL. The glucose transporter type 1 (GLUT1) expression of these cells was rapidly down-regulated in vitro. Other than GM3(NeuAc) and GM3(NeuGc) as major glycosphingolipids, PnMECs comprise GM1, GD1a, GD1b and GT1b, which are shared by PNS parenchyma, and sialyl lactosaminyl paragloboside (SLPG) as minor species. Because bovine PnMECs proliferate rapidly and a large mass of cells could be obtained, this method should contribute to the biochemical analysis of surface molecules in PnMECs that might play a key role in the formation of BNB as well as in pathological conditions involving the PNS. PMID- 9335265 TI - Lipoprotein from the osmoregulated ABC transport system OpuA of Bacillus subtilis: purification of the glycine betaine binding protein and characterization of a functional lipidless mutant. AB - The OpuA transport system of Bacillus subtilis functions as a high-affinity uptake system for the osmoprotectant glycine betaine. It is a member of the ABC transporter superfamily and consists of an ATPase (OpuAA), an integral membrane protein (OpuAB), and a hydrophilic polypeptide (OpuAC) that shows the signature sequence of lipoproteins (B. Kempf and E. Bremer, J. Biol. Chem. 270:16701-16713, 1995). The OpuAC protein might thus serve as an extracellular substrate binding protein anchored in the cytoplasmic membrane via a lipid modification at an amino terminal cysteine residue. A malE-opuAC hybrid gene was constructed and used to purify a lipidless OpuAC protein. The purified protein bound radiolabeled glycine betaine avidly and exhibited a KD of 6 microM for this ligand, demonstrating that OpuAC indeed functions as the substrate binding protein for the B. subtilis OpuA system. We have selectively expressed the opuAC gene under T7 phi10 control in Escherichia coli and have demonstrated through its metabolic labeling with [3H]palmitic acid that OpuAC is a lipoprotein. A mutant expressing an OpuAC protein in which the amino-terminal cysteine residue was changed to an alanine (OpuAC-3) was constructed by oligonucleotide site-directed mutagenesis. The OpuAC 3 protein was not acylated by [3H]palmitic acid, and part of it was secreted into the periplasmic space of E. coli, where it could be released from the cells by cold osmotic shock. The opuAC-3 mutation was recombined into an otherwise wild type opuA operon in the chromosome of B. subtilis. Unexpectedly, this mutant OpuAC system still functioned efficiently for glycine betaine acquisition in vivo under high-osmolarity growth conditions. In addition, the mutant OpuA transporter exhibited kinetic parameters similar to that of the wild-type system. Our data suggest that the lipidless OpuAC-3 protein is held in the cytoplasmic membrane of B. subtilis via its uncleaved hydrophobic signal peptide. PMID- 9335266 TI - Construction and characterization of a Bacteroides thetaiotaomicron recA mutant: transfer of Bacteroides integrated conjugative elements is RecA independent. AB - We report the construction and analysis of a Bacteroides thetaiotaomicron recA disruption mutant and an investigation of whether RecA is required for excision and integration of Bacteroides mobile DNA elements. The recA mutant was deficient in homologous recombination and was more sensitive than the wild-type strain to DNA-damaging agents. The recA mutant was also more sensitive to oxygen than the wild type, indicating that repair of DNA contributes to the aerotolerance of B. thetaiotaomicron. Many Bacteroides clinical isolates carry self-transmissible chromosomal elements known as conjugative transposons. These conjugative transposons can also excise and mobilize in trans a family of unlinked integrated elements called nonreplicating Bacteroides units (NBUs). The results of a previous study had raised the possibility that RecA plays a role in excision of Bacteroides conjugative transposons, but this hypothesis could not be tested in Bacteroides spp. because no RecA-deficient Bacteroides strain was available. We report here that the excision and integration of the Bacteroides conjugative transposons, as well as NBU1 and Tn4351, were unaffected by the absence of RecA activity. PMID- 9335267 TI - Methods for generating precise deletions and insertions in the genome of wild type Escherichia coli: application to open reading frame characterization. AB - We have developed a new system of chromosomal mutagenesis in order to study the functions of uncharacterized open reading frames (ORFs) in wild-type Escherichia coli. Because of the operon structure of this organism, traditional methods such as insertional mutagenesis run the risk of introducing polar effects on downstream genes or creating secondary mutations elsewhere in the genome. Our system uses crossover PCR to create in-frame, tagged deletions in chromosomal DNA. These deletions are placed in the E. coli chromosome by using plasmid pKO3, a gene replacement vector that contains a temperature-sensitive origin of replication and markers for positive and negative selection for chromosomal integration and excision. Using kanamycin resistance (Kn(r)) insertional alleles of the essential genes pepM and rpsB cloned into the replacement vector, we calibrated the system for the expected results when essential genes are deleted. Two poorly understood genes, hdeA and yjbJ, encoding highly abundant proteins were selected as targets for this approach. When the system was used to replace chromosomal hdeA with insertional alleles, we observed vastly different results that were dependent on the exact nature of the insertions. When a Kn(r) gene was inserted into hdeA at two different locations and orientations, both essential and nonessential phenotypes were seen. Using PCR-generated deletions, we were able to make in-frame deletion strains of both hdeA and yjbJ. The two genes proved to be nonessential in both rich and glucose-minimal media. In competition experiments using isogenic strains, the strain with the insertional allele of yjbJ showed growth rates different from those of the strain with the deletion allele of yjbJ. These results illustrate that in-frame, unmarked deletions are among the most reliable types of mutations available for wild-type E. coli. Because these strains are isogenic with the exception of their deleted ORFs, they may be used in competition with one another to reveal phenotypes not apparent when cultured singly. PMID- 9335268 TI - Functional domains of the InsA protein of IS2. AB - The InsA protein is a transcriptional regulator. It binds to the promoter region of insA and insAB'. To understand the molecular mechanism for the interaction between InsA and its binding sequence, the functional domains of InsA were identified. The glutaraldehyde cross-linking method and the two-hybrid expression system were used to study the protein-protein interaction of InsA. The results of these experiments showed that InsA forms homodimers. Deletion of the last 44 amino acid residues at its C terminus, but not the first 12 or 57 residues at the N terminus, abolished the ability of InsA to form homodimers, indicating that the protein-protein interaction domain of InsA is located at its C terminus. Gel retardation assays revealed that deletion of the last 29 amino acid residues at its C terminus had no effect on the DNA binding ability of InsA. In contrast, deletion of the first N-terminal 12 residues abolished the DNA binding capability of InsA. These results indicate that the DNA binding domain of InsA is located at its N terminus. PMID- 9335269 TI - A new Bacillus subtilis gene, med, encodes a positive regulator of comK. AB - Bacillus subtilis degR, a positive regulator of the production of degradative enzymes, is negatively regulated by the competence transcription factor ComK which is overproduced in mecA null mutants. We used transposon Tn10 to search for a mutation that reduced the repression level of degR caused by a mecA mutation. A new gene exerting positive regulation on comK was obtained and designated med (suppressor of mecA effect on degR). Sequence determination, Northern analysis, and primer extension analyses revealed that the med gene contained an open reading frame (ORF) composed of 317 codons and was transcribed into an approximately 1,250-nucleotide mRNA together with its short downstream gene. The expression of comK is positively regulated by factors such as ComK itself, ComS (SrfA)-MecA, DegU, SinR, and AbrB. Quantitative analyses using comK'-'lacZ, srfA lacZ, degU'-'lacZ, and sinR'-'lacZ fusions showed that disruption of med caused a significant decrease in comK expression in both mecA+ and mecA strains, while expression of srfA, sinR, and degU was not affected by the mutation. An epistatic analysis revealed that overproduction of ComK resulted in alteration of med expression, suggesting a regulatory loop between comK and med. Several possible mechanisms for positive regulation of comK by Med are discussed. PMID- 9335270 TI - Use of an inducible regulatory protein to identify members of a regulon: application to the regulon controlled by the leucine-responsive regulatory protein (Lrp) in Escherichia coli. AB - Procedures were developed to facilitate the identification of genes that belong to a given regulon and characterization of their responses to the regulator. The regulon controlled by the Escherichia coli leucine-responsive regulatory protein (Lrp) was studied by isolating random transcriptional fusions to lacZ, using lambda placMu53 and a strain in which lrp is under isopropylthio-beta-D galactopyranoside (IPTG)-inducible control. Fusions exhibiting IPTG-responsive beta-galactosidase activity were cloned by integrating the suicide vector pIVET1 via homologous recombination at lacZ, followed by self-ligating digested chromosomal DNA. We verified the patterns of lacZ expression after using the plasmid clones to generate merodiploid strains with interrupted and uninterrupted copies of the same sequence. If the merodiploid expression pattern was unchanged from that shown by the original fusion strain, then the cloned fusion was responsible for the regulatory pattern of interest; a difference in the expression pattern could indicate that the original strain carried multiple fusions or that there were autogenous effects of having interrupted the fused gene. Using these procedures, we generated a fusion library of approximately 5 x 10(6) strains; approximately 3,000 of these strains were screened, yielding 84 Lrp-responsive fusions, and 10 of the 84 were phenotypically stable and were characterized. The responses of different fusions in a given operon to in vivo Lrp titrations revealed variations in expression with the position of insertion. Among the newly identified members of the regulon is an open reading frame (orf3) between rpiA and serA. Also, expression of a fusion just downstream of dinF was found to be Lrp dependent only in stationary phase. PMID- 9335271 TI - Interactions of dedicated export membrane proteins of the colicin V secretion system: CvaA, a member of the membrane fusion protein family, interacts with CvaB and TolC. AB - The antibacterial peptide toxin colicin V uses a dedicated signal sequence independent system for its secretion in Escherichia coli and requires the products of three genes, cvaA, cvaB, and tolC. As a member of the membrane fusion protein family, CvaA is supposed to form a bridge that connects the inner and outer membranes via interaction with CvaB and TolC, respectively. In this study, we investigated the possible interaction of these proteins. When CvaA or CvaB was absent, the corresponding amount of CvaB or CvaA, respectively, was decreased, and the amounts of both proteins were reduced when TolC was depleted. Translational lacZ fusions showed that TolC did not affect the synthesis of either CvaA-beta-galactosidase or CvaB-beta-galactosidase, and CvaA or CvaB did not affect the synthesis of CvaB-beta-galactosidase or CvaA-beta-galactosidase, respectively. However, the stabilities of CvaA and CvaB proteins were affected by the absence of one another and by that of TolC. The instability of CvaA was more severe in TolC-depleted cells than in CvaB-depleted cells. On the other hand, CvaB was less stable in the absence of CvaA than in the absence of TolC. In addition, using a cross-linking reagent, we showed that CvaA directly interacts with both CvaB and TolC proteins. Taken together, these data support the hypothesized structural role of CvaA in connecting CvaB and TolC. PMID- 9335272 TI - Cloning of an avilamycin biosynthetic gene cluster from Streptomyces viridochromogenes Tu57. AB - A 65-kb region of DNA from Streptomyces viridochromogenes Tu57, containing genes encoding proteins involved in the biosynthesis of avilamycins, was isolated. The DNA sequence of a 6.4-kb fragment from this region revealed four open reading frames (ORF1 to ORF4), three of which are fully contained within the sequenced fragment. The deduced amino acid sequence of AviM, encoded by ORF2, shows 37% identity to a 6-methylsalicylic acid synthase from Penicillium patulum. Cultures of S. lividans TK24 and S. coelicolor CH999 containing plasmids with ORF2 on a 5.5-kb PstI fragment were able to produce orsellinic acid, an unreduced version of 6-methylsalicylic acid. The amino acid sequence encoded by ORF3 (AviD) is 62% identical to that of StrD, a dTDP-glucose synthase from S. griseus. The deduced amino acid sequence of AviE, encoded by ORF4, shows 55% identity to a dTDP glucose dehydratase (StrE) from S. griseus. Gene insertional inactivation experiments of aviE abolished avilamycin production, indicating the involvement of aviE in the biosynthesis of avilamycins. PMID- 9335273 TI - Altered extent of cross-linking of beta1,6-glucosylated mannoproteins to chitin in Saccharomyces cerevisiae mutants with reduced cell wall beta1,3-glucan content. AB - The yeast cell wall contains beta1,3-glucanase-extractable and beta1,3-glucanase resistant mannoproteins. The beta1,3-glucanase-extractable proteins are retained in the cell wall by attachment to a beta1,6-glucan moiety, which in its turn is linked to beta1,3-glucan (J. C. Kapteyn, R. C. Montijn, E. Vink, J. De La Cruz, A. Llobell, J. E. Douwes, H. Shimoi, P. N. Lipke, and F. M. Klis, Glycobiology 6:337-345, 1996). The beta1,3-glucanase-resistant protein fraction could be largely released by exochitinase treatment and contained the same set of beta1,6 glucosylated proteins, including Cwp1p, as the B1,3-glucanase-extractable fraction. Chitin was linked to the proteins in the beta1,3-glucanase-resistant fraction through a beta1,6-glucan moiety. In wild-type cell walls, the beta1,3 glucanase-resistant protein fraction represented only 1 to 2% of the covalently linked cell wall proteins, whereas in cell walls of fks1 and gas1 deletion strains, which contain much less beta1,3-glucan but more chitin, beta1,3 glucanase-resistant proteins represented about 40% of the total. We propose that the increased cross-linking of cell wall proteins via beta1,6-glucan to chitin represents a cell wall repair mechanism in yeast, which is activated in response to cell wall weakening. PMID- 9335274 TI - Dual role of alpha-acetolactate decarboxylase in Lactococcus lactis subsp. lactis. AB - The alpha-acetolactate decarboxylase gene aldB is clustered with the genes for the branched-chain amino acids (BCAA) in Lactococcus lactis subsp. lactis. It can be transcribed with BCAA genes under isoleucine regulation or independently of BCAA synthesis under the control of its own promoter. The product of aldB is responsible for leucine sensibility under valine starvation. In the presence of more than 10 microM leucine, the alpha-acetolactate produced by the biosynthetic acetohydroxy acid synthase IlvBN is transformed to acetoin by AldB and, consequently, is not available for valine synthesis. AldB is also involved in acetoin formation in the 2,3-butanediol pathway, initiated by the catabolic acetolactate synthase, AlsS. The differences in the genetic organization, the expression, and the kinetics parameters of these enzymes between L. lactis and Klebsiella terrigena, Bacillus subtilis, or Leuconostoc oenos suggest that this pathway plays a different role in the metabolism in these bacteria. Thus, the alpha-acetolactate decarboxylase from L. lactis plays a dual role in the cell: (i) as key regulator of valine and leucine biosynthesis, by controlling the acetolactate flux by a shift to catabolism; and (ii) as an enzyme catalyzing the second step of the 2,3-butanediol pathway. PMID- 9335275 TI - Increase of methicillin resistance in Staphylococcus aureus caused by deletion of a gene whose product is homologous to lytic enzymes. AB - A spontaneous high-level methicillin-resistant mutant, SRM1648, for which the MIC of methicillin is 1,600 microg/ml, was isolated on a plate containing 400 microg of the antibiotic/ml on which had been cultured the low-level methicillin resistant Staphylococcus aureus SR17238, for which the MIC is 6.3 microg/ml. Analysis of the chromosomal DNAs of the mutant and the parental strains by the restriction landmark genomic scanning method with two-dimensional electrophoresis of restriction fragments revealed a 1.6-kb deletion in the chromosome of the mutant. The HindIII fragment of 2.5 kb containing this deleted region was cloned into a plasmid vector and introduced into the parental strain. A deletion mutant reconstructed in the presence of a low concentration of methicillin by integration and excision of the recombinant plasmid exhibited a high level of resistance (methicillin MIC, 1,600 microg/ml), confirming that the deletion had caused the elevation of the resistance level. Sequence analysis indicated that the deletion occurred in three consecutive open reading frames (ORFs). The predicted amino acid sequence of the first ORF showed high homology with both RelA and SpoT of Escherichia coli, which are involved in the synthesis and hydrolysis of guanosine 5',3'-polyphosphate, and that of the third ORF showed a relatively high homology to the lytic enzyme encoded by the lytC gene of Bacillus subtilis. We also isolated another high-level resistant mutant with a deletion within the third ORF, which suggested that inactivation of some lytic enzyme resulted in the increased resistance. PMID- 9335276 TI - Bacillus subtilis PhoP binds to the phoB tandem promoter exclusively within the phosphate starvation-inducible promoter. AB - Several gene products, including three two-component systems, make up a signal transduction network that controls the phosphate starvation response in Bacillus subtilis. Epistasis experiments indicate that PhoP, a response regulator, is furthest downstream of the known regulators in the signaling pathway that regulates Pho regulon genes. We report the overexpression, purification, and use of PhoP in investigating its role in Pho regulon gene activation. PhoP was a substrate for both the kinase and phosphatase activities of its cognate sensor kinase, PhoR. It was not phosphorylated by acetyl phosphate. Purified phosphorylated PhoP (PhoPP) had a half-life of approximately 2.5 h, which was reduced to about 15 min by addition of the same molar amount of *PhoR (the cytoplasmic region of PhoR). ATP significantly increased phosphatase activity of *PhoR on PhoPP. In gel filtration and cross-linking studies, both PhoP and PhoPP were shown to be dimers. The dimerization domain was located within the 135 amino acids at the N terminus of PhoP. Phosphorylated or unphosphorylated PhoP bound to one of the alkaline phosphatase gene promoters, the phoB promoter. Furthermore, PhoP bound exclusively to the -18 to -73 region (relative to the transcriptional start site +1) of the phosphate starvation-inducible promoter (Pv) but not to the adjacent developmentally regulated promoter (Ps). These data corroborate the genetic data for phoB regulation and suggest that activation of phoB is via direct interaction between PhoP and the phoB promoter. Studies of the phosphorylation, oligomerization, and DNA binding activity of the PhoP protein demonstrate that its N-terminal phosphorylation and dimerization domain and its C terminal DNA binding domain function independently of one another, distinguishing PhoP from other response regulators, such as PhoB (Escherichia coli) and NtrC. PMID- 9335277 TI - Characterization of the oriC region of Mycobacterium smegmatis. AB - A 3.5-kb DNA fragment containing the dnaA region of Mycobacterium smegmatis has been hypothesized to be the chromosomal origin of replication or oriC (M. Rajagopalan et al., J. Bacteriol. 177:6527-6535, 1995). This region included the rpmH gene, the dnaA gene, and a major portion of the dnaN gene as well as the rpmH-dnaA and dnaA-dnaN intergenic regions. Deletion analyses of this region revealed that a 531-bp DNA fragment from the dnaA-dnaN intergenic region was sufficient to exhibit oriC activity, while a 495-bp fragment from the same region failed to exhibit oriC activity. The oriC activities of plasmids containing the 531-bp sequence was less than the activities of those containing the entire dnaA region, suggesting that the regions flanking the 531-bp sequence stimulated oriC activity. The 531-bp region contained several putative nine-nucleotide DnaA protein recognition sequences [TT(G/C)TCCACA] and a single 11-nucleotide AT-rich cluster. Replacement of adenine with guanine at position 9 in five of the putative DnaA boxes decreased oriC activity. Mutations at other positions in two of the DnaA boxes also decreased oriC activity. Deletion of the 11-nucleotide AT rich cluster completely abolished oriC activity. These data indicate that the designated DnaA boxes and the AT-rich cluster of the M. smegmatis dnaA-dnaN intergenic region are essential for oriC activity. We suggest that M. smegmatis oriC replication could involve interactions of the DnaA protein with the putative DnaA boxes as well as with the AT-rich cluster. PMID- 9335278 TI - Characterization of two heat shock genes from Haloferax volcanii: a model system for transcription regulation in the Archaea. AB - The expression of two heat-responsive cct (chaperonin-containing Tcp-1) genes from the archaeon Haloferax volcanii was investigated at the transcription level. The cct1 and cct2 genes, which encode proteins of 560 and 557 amino acids, respectively, were identified on cosmid clones of an H. volcanii genomic library and subsequently sequenced. The deduced amino acid sequences of these genes exhibited a high degree of similarity to other archaeal and eucaryal cct family members. Expression of the cct genes was characterized in detail for the purpose of developing a model for studying transcription regulation in the domain Archaea. Northern (RNA) analysis demonstrated that the cct mRNAs were maximally induced after heat shock from 37 to 55 degrees C and showed significant heat inducibility after 30 min at 60 degrees C. Transcription of cct mRNAs was also stimulated in response to dilute salt concentrations. Transcriptional analysis of cct promoter regions coupled to a yeast tRNA reporter gene demonstrated that 5' flanking sequences up to position -233 (cct1) and position -170 (cct2) were sufficient for promoting heat-induced transcription. Transcript analysis indicated that both basal transcription and stress-induced transcription of the H. volcanii cct genes were directed by a conserved archaeal consensus TATA motif (5'-TTTATA-3') centered at -25 relative to the mapped initiation site. Comparison of the cct promoter regions also revealed a striking degree of sequence conservation immediately 5' and 3' of the TATA element. PMID- 9335279 TI - Rapamycin specifically interferes with the developmental response of fission yeast to starvation. AB - Rapamycin is a microbial macrolide which belongs to a family of immunosuppressive drugs that suppress the immune system by blocking stages of signal transduction in T lymphocytes. In Saccharomyces cerevisiae cells, as in T lymphocytes, rapamycin inhibits growth and cells become arrested at the G1 stage of the cell cycle. Rapamycin is also an effective antifungal agent, affecting the growth of yeast and filamentous fungi. Unexpectedly, we observed that rapamycin has no apparent effect on the vegetative growth of Schizosaccharomyces pombe. Instead, the drug becomes effective only when cells experience starvation. Under such conditions, homothallic wild-type cells will normally mate and undergo sporulation. In the presence of rapamycin, this sexual development process is strongly inhibited and cells adopt an alternative physiological option and enter stationary phase. Rapamycin strongly inhibits sexual development of haploid cells prior to the stage of sexual conjugation. In contrast, the drug has only a slight inhibitory effect on the sporulation of diploid cells. A genetic approach was applied to identify the signal transduction pathway that is inhibited by rapamycin. The results indicate that either rapamycin did not suppress the derepression of sexual development of strains in which adenylate cyclase was deleted or the cyclic AMP-dependent protein kinase encoded by pka1 was mutated. Nor did rapamycin inhibit the unscheduled meiosis observed in pat1-114 mutants. Overexpression of ras1+, an essential gene for sexual development, did not rescue the sterility of rapamycin-treated cells. However, expression of the activated allele, ras1Val17, antagonized the effect of rapamycin and restored the ability of the cells to respond to mating signals in the presence of the drug. We discuss possible mechanisms for the inhibitory effect of rapamycin on sexual development in S. pombe. PMID- 9335280 TI - Cloning, nucleotide sequence, and overexpression of smoS, a component of a novel operon encoding an ABC transporter and polyol dehydrogenases of Rhodobacter sphaeroides Si4. AB - The gene coding for sorbitol dehydrogenase (SDH) of Rhodobacter sphaeroides Si4 was located 55 nucleotides upstream of the mannitol dehydrogenase gene (mtlK) within a previously unrecognized polyol operon. This operon probably consists of all the proteins necessary for transport and metabolization of various polyols. The gene encoding SDH (smoS) was cloned and sequenced. Analysis of the deduced amino acid sequence revealed homology to enzymes of the short-chain dehydrogenase/reductase protein family. For structure analysis of this unique bacterial enzyme, smoS was subcloned into the overexpression vector pET-24a(+) and then overproduced in Escherichia coli BL21(DE3), which yielded a specific activity of 24.8 U/mg of protein and a volumetric yield of 38,000 U/liter. Compared to values derived with the native host, R. sphaeroides, these values reflected a 270-fold increase in expression of SDH and a 971-fold increase in the volumetric yield. SDH was purified to homogeneity, with a recovery of 49%, on the basis of a three-step procedure. Upstream from smoS, another gene (smoK), which encoded a putative ATP-binding protein of an ABC transporter, was identified. PMID- 9335281 TI - Mechanism of decay of the cry1Aa mRNA in Bacillus subtilis. AB - We undertook the study of the decay process of the cry1Aa mRNA of Bacillus thuringiensis expressed in B. subtilis. The cry1Aa transcript is a 3.7-kb mRNA expressed during sporulation whose transcriptional control has previously been studied in both B. subtilis and B. thuringiensis. We found that the cry1Aa mRNA has a half-life of around 9 min and that its decay occurs through endoribonucleolytic cleavages which result in three groups of high-molecular weight mRNA intermediates ranging in size from 2.7 to 0.5 kb. A comparative study carried out with Escherichia coli showed a similar pattern of degradation intermediates. Primer extension analysis carried out on RNA from B. subtilis revealed that most cleavages occur within two regions located toward the 5' and 3' ends of the mRNA. The most prominent processing site observed for the cry1Aa mRNA isolated from B. subtilis is only two bases away from that occurring on RNA isolated from E. coli. Most cleavage sites occur at seemingly single-stranded RNA segments rich in A and U nucleotides, suggesting that a common and conserved mechanism may process the cry1Aa mRNA. PMID- 9335282 TI - Factors controlling in vitro recrystallization of the Caulobacter crescentus paracrystalline S-layer. AB - The S-layer of Caulobacter is a two-dimensional paracrystalline array on the cell surface composed of a single protein, RsaA. We have established conditions for preparation of stable, soluble protein and then efficient in vitro recrystallization of the purified protein. Efficient recrystallization and long range order could not be obtained with pure protein only, though it was apparent that calcium was required for crystallization. Recrystallization was obtained when lipid vesicles were provided, but only when the vesicles contained the specific species of Caulobacter smooth lipopolysaccharide (SLPS) that previous studies implicated as a requirement for attaching the S-layer to the cell surface. The specific type of phospholipids did not appear critical; phospholipids rather different from those present in Caulobacter membranes or archaebacterial tetraether lipids worked equally well. The source of LPS was critical; rough and smooth variants of Salmonella typhimurium LPS as well as the rough form of Caulobacter LPS were ineffective. The requirement for calcium ions for recrystallization was further evaluated; strontium ions could substitute for calcium, and to a lesser extent, cobalt, barium, manganese and magnesium ions also stimulated crystallization. On the other hand, nickel and cadmium provided only weak crystallization stimulation, and zinc, copper, iron, aluminum ions, and the monovalent potassium, sodium, and lithium ions were ineffective. The recrystallization could also be reproduced with Langmuir-Blodgett lipid monolayers at an air-water interface. As with the vesicle experiments, this was only successful when SLPS was incorporated into the lipid mix. The best method for RsaA preparation, leading to apparently monomeric protein that was stable for many months, was an extraction with a low pH aqueous solution. We also achieved recrystallization, albeit at lower efficiency, using RsaA protein solubilized by 8 M urea, a method which allows retrieval of protein from inclusions, when expressed as heterologous protein in Escherichia coli or when retrieved as shed, precipitated protein from certain mutant caulobacters. In summary, the clarification of recrystallization methods has confirmed the requirement of SLPS as a surface attachment component and suggests that its presence in a membrane like structure greatly stimulates the extent and quality of S-layer formation. The in vitro approach allowed the demonstration that specific ions are capable of participating in crystallization and now provides an assay for the crystallization potential of modified S-layer proteins, whether they were produced in or can be secreted by caulobacters. PMID- 9335283 TI - Alternative transcription factor sigmaSB of Staphylococcus aureus: characterization and role in transcription of the global regulatory locus sar. AB - A homolog of the multiple-stress-responsive transcription factor sigmaB of Bacillus subtilis was predicted from the DNA sequence analysis of a region of the Staphylococcus aureus chromosome. A hybrid between the coding sequence of the first 11 amino acids of the gene 10 leader peptide of phage T7 (T7.Tag) and the putative sigB gene of S. aureus was constructed and cloned into Escherichia coli BL21(DE3)pLysS for overexpression from a T7 promoter. A homogeneous preparation of the overproduced protein was obtained by affinity chromatography with a T7.Tag monoclonal antibody coupled to agarose. The amino-terminal amino acid sequence of the first 22 residues of the purified protein matched that deduced from the nucleotide sequence. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified protein, designated sigmaSB, indicated that it migrated as an approximately 39-kDa polypeptide. Promoter-specific transcription from the B. subtilis sigmaB-dependent PB promoter of the sigB operon was stimulated by sigmaSB in a concentration-dependent fashion when reconstituted with the S. aureus core RNA polymerase (RNAP). Specific transcript from the predicted sigmaB dependent PB promoter of the sigB operon of S. aureus was obtained by the reconstituted RNAP in a runoff transcription reaction. The sar operon of S. aureus contains three promoter elements (P1, P2, and P3) and is known to partly control the synthesis of a number of extracellular toxins and several cell wall proteins. Our in vitro studies revealed that transcription from the P1 promoter is dependent on the primary sigma factor sigmaSA, while that of the P3 promoter is dependent on sigmaSB. As determined by primer extension studies, the 5' end of the sigmaSB-initiated mRNA synthesized in vitro from the sar P3 promoter is in agreement with the 5' end of the cellular RNA. PMID- 9335284 TI - The presence of a dnaK (HSP70) multigene family in members of the orders Planctomycetales and Verrucomicrobiales. AB - Sequences of the dnaK gene, coding for the 70-kDa heat shock protein (HSP70), were determined for six members of the order Planctomycetales, including representatives of three genera, and for the only cultivated member of the order Verrucomicrobiales, Verrucomicrobium spinosum. A fragment of the dnaK gene was amplified from these strains by PCR with oligonucleotide primers targeting regions of the dnaK gene that are conserved at the amino acid level, and the resulting PCR products were cloned into a plasmid vector. Sequence analysis of the cloned dnaK fragments revealed the presence of two different types of dnaK sequence in one of the planctomycete strains, Planctomyces maris, and in V. spinosum. Only one type of dnaK sequence was found for each of the remaining strains. Phylogenetic analysis of the partial sequence data suggested that the majority of planctomycete strains, including one of the Planctomyces maris sequences, form a coherent phylogenetic group branching adjacent to other main lines of descent within the domain Bacteria, as has been shown previously by 16S rRNA sequence analysis. One of the two V. spinosum dnaK sequences also appears to constitute a separate lineage within the gram-negative bacteria. Each of the remaining sequences from P. maris and V. spinosum, together with the single sequence obtained from Planctomyces limnophilus, appeared to be unrelated to the other planctomycete sequences and to occupy a position distant from that of other gram-negative bacteria. The phylogenetic diversity of dnaK sequences exhibited by P. maris and V. spinosum was comparable to that found in Synechococcus sp. strain PCC7942 and Escherichia coli, the only other prokaryotes for which a dnaK multigene family has been demonstrated. PMID- 9335285 TI - Deletion analysis of the fis promoter region in Escherichia coli: antagonistic effects of integration host factor and Fis. AB - Fis is a small DNA-binding and -bending protein in Escherichia coli that is involved in several different biological processes, including stimulation of specialized DNA recombination events and regulation of gene expression. fis protein and mRNA levels rapidly increase during early logarithmic growth phase in response to a nutritional upshift but become virtually undetectable during late logarithmic and stationary phases. We present evidence that the growth phase dependent fis expression pattern is not determined by changes in mRNA stability, arguing in favor of regulation at the level of transcription. DNA deletion analysis of the fis promoter (fis P) region indicated that DNA sequences from 166 to -81, -36 to -26, and +107 to +366 relative to the transcription start site are required for maximum expression. A DNA sequence resembling the integration host factor (IHF) binding site centered approximately at -114 showed DNase I cleavage protection by IHF. In ihf cells, maximum cellular levels of fis mRNA were decreased more than 3-fold and transcription from fis P on a plasmid was decreased about 3.8-fold compared to those in cells expressing wild-type IHF. In addition, a mutation in the ihf binding site resulted in a 76 and 61% reduction in transcription from fis P on a plasmid in the presence or absence of Fis, respectively. Insertions of 5 or 10 bp between this ihf site and fis P suggest that IHF functions in a position-dependent manner. We conclude that IHF plays a role in stimulating transcription from fis P by interacting with a site centered approximately at -114 relative to the start of transcription. We also showed that although the fis P region contains six Fis binding sites, Fis site II (centered at -42) played a predominant role in autoregulation, Fis sites I and III (centered at +26 and -83, respectively) seemingly played smaller roles, and no role in negative autoregulation could be attributed to Fis sites IV, V, and VI (located upstream of site III). The fis P region from -36 to +7, which is not directly regulated by either IHF or Fis, retained the characteristic fis regulation pattern in response to a nutritional upshift. PMID- 9335286 TI - The Mycobacterium xenopi GyrA protein splicing element: characterization of a minimal intein. AB - The 198-amino-acid in-frame insertion in the gyrA gene of Mycobacterium xenopi is the smallest known naturally occurring active protein splicing element (intein). Comparison with other mycobacterial gyrA inteins suggests that the M. xenopi intein underwent a complex series of events including (i) removal of 222 amino acids that encompass most of the central intein domain, and (ii) addition of a linker of unrelated residues. This naturally occurring genetic rearrangement is a representative characteristic of the taxon. The deletion process removes the conserved motifs involved in homing endonuclease activity. The linker insertion represents a structural requirement, as its mutation resulted in failure to splice. The M. xenopi GyrA intein thus provides a paradigm for a minimal protein splicing element. PMID- 9335287 TI - Amylase and chitinase genes in Streptomyces lividans are regulated by reg1, a pleiotropic regulatory gene. AB - A regulatory gene, reg1, was identified in Streptomyces lividans. It encodes a 345-amino-acid protein (Reg1) which contains a helix-turn-helix DNA-binding motif in the N-terminal region. Reg1 exhibits similarity with the LacI/GalR family members over the entire sequence. It displays 95% identity with MalR (the repressor of malE in S. coelicolor), 65% identity with ORF-Sl (a putative regulatory gene of alpha-amylase of S. limosus), and 31% identity with CcpA (the carbon catabolite repressor in Bacillus subtilis). In S. lividans, the chromosomal disruption of reg1 affected the expression of several genes. The production of alpha-amylases of S. lividans and that of the alpha-amylase of S. limosus in S. lividans were enhanced in the reg1 mutant strains and relieved of carbon catabolite repression. As a result, the transcription level of the alpha amylase of S. limosus was noticeably increased in the reg1 mutant strain. Moreover, the induction of chitinase production in S. lividans was relieved of carbon catabolite repression by glucose in the reg1 mutant strain, while the induction by chitin was lost. Therefore, reg1 can be regarded as a pleiotropic regulatory gene in S. lividans. PMID- 9335288 TI - Three genes of a motility operon and their role in flagellar rotary speed variation in Rhizobium meliloti. AB - The peritrichous flagella of Rhizobium meliloti rotate only clockwise and control directional changes of swimming cells by modulating flagellar rotary speed. Using Tn5 insertions, we have identified and sequenced a motility (mot) operon containing three genes, motB, motC, and motD, that are translationally coupled. The motB gene (and an unlinked motA) has been assigned by similarity to the Escherichia coli and Bacillus subtilis homologs, whereas motC and motD are new and without known precedents in other bacteria. In-frame deletions introduced in motB, motC, or motD each result in paralysis. MotD function was fully restored by complementation with the wild-type motD gene. By contrast, deletions in motB or motC required the native combination of motB and motC in trans for restoring normal flagellar rotation, whereas complementation with motB or motC alone led to uncoordinated (jiggly) swimming. Similarly, a motB-motC gene fusion and a Tn5 insertion intervening between motB and motC resulted in jiggly swimming as a consequence of large fluctuations in flagellar rotary speed. We conclude that MotC biosynthesis requires coordinate expression of motB and motC and balanced amounts of the two gene products. The MotC polypeptide contains an N-terminal signal sequence for export, and Western blots have confirmed its location in the periplasm of the R. meliloti cell. A working model suggests that interactions between MotB and MotC at the periplasmic surface of the motor control the energy flux or the energy coupling that drives flagellar rotation. PMID- 9335289 TI - Identification of human transferrin-binding sites within meningococcal transferrin-binding protein B. AB - Transferrin-binding protein B (TbpB) from Neisseria meningitidis binds human transferrin (hTf) at the surface of the bacterial cell as part of the iron uptake process. To identify hTf binding sites within the meningococcal TbpB, defined regions of the molecule were produced in Escherichia coli by a translational fusion expression system and the ability of the recombinant proteins (rTbpB) to bind peroxidase-conjugated hTf was characterized by Western blot and dot blot assays. Both the N-terminal domain (amino acids [aa] 2 to 351) and the C-terminal domain (aa 352 to 691) were able to bind hTf, and by a peptide spot synthesis approach, two and five hTf binding sites were identified in the N- and C-terminal domains, respectively. The hTf binding activity of three rTbpB deletion variants constructed within the central region (aa 346 to 543) highlighted the importance of a specific peptide (aa 377 to 394) in the ligand interaction. Taken together, the results indicated that the N- and C-terminal domains bound hTf approximately 10 and 1000 times less, respectively, than the full-length rTbpB (aa 2 to 691), while the central region (aa 346 to 543) had a binding avidity in the same order of magnitude as the C-terminal domain. In contrast with the hTf binding in the N terminal domain, which was mediated by conformational epitopes, linear determinants seemed to be involved in the hTf binding in the C-terminal domain. The host specificity for transferrin appeared to be mediated by the N-terminal domain of the meningococcal rTbpB rather than the C-terminal domain, since we report that murine Tf binds to the C-terminal domain. Antisera raised to both N- and C-terminal domains were bactericidal for the parent strain, indicating that both domains are accessible at the bacterial surface. We have thus identified hTf binding sites within each domain of the TbpB from N. meningitidis and propose that the N- and C-terminal domains together contribute to the efficient binding of TbpB to hTf with their respective affinities and specificities for determinants of their ligand. PMID- 9335290 TI - Characterization of two genes encoding the Mycobacterium tuberculosis ribonucleotide reductase small subunit. AB - Two nrdF genes, nrdF1 and nrdF2, encoding the small subunit (R2) of ribonucleotide reductase (RR) from Mycobacterium tuberculosis have 71% identity at the amino acid level and are both highly homologous with Salmonella typhimurium R2F. The calculated molecular masses of R2-1 and R2-2 are 36,588 (322 amino acids [aa]) and 36,957 (324 aa) Da, respectively. Western blot analysis of crude M. tuberculosis extracts indicates that both R2s are expressed in vivo. Recombinant R2-2 is enzymatically active when assayed with pure recombinant M. tuberculosis R1 subunit. Both ATP and dATP are activators for CDP reduction up to 2 and 1 mM, respectively. The gene encoding M. tuberculosis R2-1, nrdF1, is not linked to nrdF2, nor is either gene linked to the gene encoding the large subunit, M. tuberculosis nrdE. The gene encoding MTP64 was found downstream from nrdF1, and the gene encoding alcohol dehydrogenase was found downstream from nrdF2. A nrdA(Ts) strain of E. coli (E101) could be complemented by simultaneous transformation with M. tuberculosis nrdE and nrdF2. An M. tuberculosis nrdF2 variant in which the codon for the catalytically necessary tyrosine was replaced by the phenylalanine codon did not complement E101 when cotransformed with M. tuberculosis nrdE. Similarly, M. tuberculosis nrdF1 and nrdE did not complement E101. Activity of recombinant M. tuberculosis RR was inhibited by incubating the enzyme with a peptide corresponding to the 7 C-terminal amino acid residues of the R2-2 subunit. M. tuberculosis is a species in which a nrdEF system appears to encode the biologically active species of RR and also the only bacterial species identified so far in which class I RR subunits are not arranged on an operon. PMID- 9335291 TI - Acyltransferase domain substitutions in erythromycin polyketide synthase yield novel erythromycin derivatives. AB - The methylmalonyl coenzyme A (methylmalonyl-CoA)-specific acyltransferase (AT) domains of modules 1 and 2 of the 6-deoxyerythronolide B synthase (DEBS1) of Saccharopolyspora erythraea ER720 were replaced with three heterologous AT domains that are believed, based on sequence comparisons, to be specific for malonyl-CoA. The three substituted AT domains were "Hyg" AT2 from module 2 of a type I polyketide synthase (PKS)-like gene cluster isolated from the rapamycin producer Streptomyces hygroscopicus ATCC 29253, "Ven" AT isolated from a PKS-like gene cluster of the pikromycin producer Streptomyces venezuelae ATCC 15439, and RAPS AT14 from module 14 of the rapamycin PKS gene cluster of S. hygroscopicus ATCC 29253. These changes led to the production of novel erythromycin derivatives by the engineered strains of S. erythraea ER720. Specifically, 12-desmethyl-12 deoxyerythromycin A, which lacks the methyl group at C-12 of the macrolactone ring, was produced by the strains in which the resident AT1 domain was replaced, and 10-desmethylerythromycin A and 10-desmethyl-12-deoxyerythromycin A, both of which lack the methyl group at C-10 of the macrolactone ring, were produced by the recombinant strains in which the resident AT2 domain was replaced. All of the novel erythromycin derivatives exhibited antibiotic activity against Staphylococcus aureus. The production of the erythromycin derivatives through AT replacements confirms the computer predicted substrate specificities of "Hyg" AT2 and "Ven" AT and the substrate specificity of RAPS AT14 deduced from the structure of rapamycin. Moreover, these experiments demonstrate that at least some AT domains of the complete 6-deoxyerythronolide B synthase of S. erythraea can be replaced by functionally related domains from different organisms to make novel, bioactive compounds. PMID- 9335292 TI - Identification of an essential Caulobacter crescentus gene encoding a member of the Obg family of GTP-binding proteins. AB - We have identified an essential Caulobacter crescentus gene (cgtA) that encodes a member of a recently identified subfamily of GTPases (the Obg family) conserved from Bacteria to Archaea to humans. This evolutionary conservation between distantly related species suggests that this family of GTP-binding proteins possesses a fundamental, yet unknown, cellular role. In this report, we describe the isolation and sequence of the cgtA gene. The predicted CgtA protein displays striking similarity to the Obg family of small, monomeric GTP-binding proteins, both in the conserved guanine nucleotide-binding domains and throughout the N terminal glycine-rich domain that is found in many members of the Obg family. Disruption of the cgtA gene was lethal, demonstrating that this gene is essential for cell growth. Immunoblot analysis revealed that CgtA protein levels remained constant throughout the C. crescentus cell cycle. PMID- 9335293 TI - Stationary-phase mutants of Sinorhizobium meliloti are impaired in stationary phase survival or in recovery to logarithmic growth. AB - A screening method was used to identify Sinorhizobium meliloti mutants which are affected in stationary-phase survival. Of 20,000 individual colonies mutagenized with transposon Tn5-B20, 10 mutant strains which showed poor or no survival in the stationary phase were identified. Analyses of expression patterns of the promoterless lacZ genes in the mutant strains revealed individual induction patterns. Most strains were induced in stationary phase as well as under carbon limitation and in pure H2O, but none of the mutants was induced under heat, alkali stress conditions, or low oxygen tension. Plant inoculation tests revealed that the symbiotic proficiency of the mutants was not affected. Two mutants, however, showed gene induction not only in the stationary phase under free-living conditions but also in the bacteroid state. A long-term starvation test was carried out to examine the ability of the 10 mutants to survive prolonged stationary-phase conditions. All mutants showed a clear decrease in the colony forming ability under the chosen experimental conditions. Staining with green and red fluorescent nucleic acid stain showed that the mutants fell into two different classes. Seven mutants died during stationary phase; the three other mutants remained viable but did not resume growth after prolonged starvation. Five of the ten Tn5-B20 insertions were cloned from the genomes of the mutant strains. Nucleotide sequence analyses established that the transposon had inserted in five distinctive genes. Database searches revealed that four of the tagged loci corresponded to already characterized genes whose gene products are involved in important cellular processes such as amino acid metabolism or aerobic respiration. PMID- 9335294 TI - Identification, isolation, and characterization of the 42-kilodalton major outer membrane protein (MompA) from Treponema pectinovorum ATCC 33768. AB - The major protein present in the isolated outer membrane of Treponema pectinovorum ATCC 33768, MompA, was identified, purified, and characterized. Immuno-gold electron microscopy, using anti-MompA serum, and cell fractionation experiments confirmed the localization of MompA to the outer membrane. MompA was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to have a molecular mass of 42 kDa when heat denatured, whereas native MompA formed a number of detergent-stable forms with molecular masses of 71, 76, and 83 kDa. A temperature of 60 degrees C was required to convert the native protein to the 42 kDa form. A number of detergents and chemical agents that are capable of breaking ionic and hydrogen bonds of proteins did not convert native MompA to the 42-kDa species. The native forms of the protein were resistant to the combined action of proteinase K, trypsin, and chymotrypsin, whereas the 42-kDa form of MompA was not. The N-terminal amino acid sequence of MompA was determined to be DVTVNINSRVRPVLYTT, and database searches did not identify any homology with known protein sequences. Amino acid compositional analysis showed the protein to be rich in proline and glycine, with these amino acids accounting for 28 and 13%, respectively, of the total amino acids. Antiserum raised against the major outer membrane protein of T. denticola GM-1 and ATCC 35405 did not cross-react with MompA, and antiserum raised against MompA did not react with any cellular components of Treponema denticola, Treponema vincentii, or Treponema socranskii. A major outer membrane protein similar in molecular mass to MompA was identified in eight clinical isolates of T. pectinovorum. The major outer membrane protein produced by four of the clinical isolates reacted strongly, by Western blotting, with anti-MompA serum, whereas proteins of the other strains did not. PMID- 9335295 TI - Transcriptional characterization of the Rickettsia prowazekii major macromolecular synthesis operon. AB - Recent studies have demonstrated that Rickettsia prowazekii can regulate transcription of selected genes at the level of initiation. However, little information concerning the existence of operons and coordinate gene regulation in this obligate intracellular parasitic bacterium is available. To address these issues, we have focused on the rpoD gene linkage group (greA-open reading frame 23 [ORF23]-dnaG-rpoD), which includes the rickettsial analog (ORF23-dnaG-rpoD) of the major macromolecular synthesis operon (MMSO). The rickettsial MMSO consists of an ORF coding for a protein of unknown function the structural genes for DNA primase (dnaG) and the major sigma factor of RNA polymerase (rpoD). RNase protection assays (RPA) were used to determine if these genes are organized into an operon controlled by multiple promoters and the quantities of transcripts produced by these genes relative to each other. RPA with a probe spanning the 270 base greA-ORF23 intervening region identified a putative transcriptional promoter within the intervening sequence. Multiple RPA probes spanning the next 4,041 bases of the linkage group demonstrated the presence of a continuous transcript and thus the existence of an operon. A probe spanning the dnaG-rpoD region revealed that two additional mRNA fragments were also protected, which enabled us to identify additional putative promoters for rpoD within dnaG. Primer extension determined that the 5' ends of the three transcripts consist separately of adenine (located 227 bases upstream of ORF23) and uracil and adenine (located 336 and 250 bases upstream of rpoD, respectively). Quantitation of transcripts produced by the three ORFs determined the relative amounts of transcripts (ORF23 to dnaG to rpoD) to be 1:2.7:5.1. PMID- 9335296 TI - Structural characterization of the lipid A component of Helicobacter pylori rough and smooth-form lipopolysaccharides. AB - The chemical structure of free lipid A isolated from rough- and smooth-form lipopolysaccharides (R-LPS and S-LPS, respectively) of the human gastroduodenal pathogen Helicobacter pylori was elucidated by compositional and degradative analysis, nuclear magnetic resonance spectroscopy, and mass spectrometry. The predominant molecular species in both lipid A components are identical and tetraacylated, but a second molecular species which is hexaacylated is also present in lipid A from S-LPS. Despite differences in substitution by acyl chains, the hydrophilic backbone of the molecules consisted of beta(1,6)-linked D glucosamine (GlcN) disaccharide 1-phosphate. Because of microheterogeneity, nonstoichiometric amounts of ethanolamine-phosphate were also linked to the glycosidic hydroxyl group. In S-LPS, but not in R-LPS, the hydroxyl group at position 4' was partially substituted by another phosphate group. Considerable variation in the distribution of fatty acids on the lipid A backbone was revealed by laser desorption mass spectrometry. In tetraacyl lipid A, the amino group of the reducing GlcN carried (R)-3-hydroxyoctadecanoic acid (position 2), that of the nonreducing GlcN carried (R)-3-(octadecanoyloxy)octadecanoic acid (position 2'), and ester-bound (R)-3-hydroxyhexadecanoic acid was attached at position 3. Hexaacyl lipid A had a similar substitution by fatty acids, but in addition, ester-bound (R)-3-(dodecanoyloxy)hexadecanoic acid or (R) 3(tetradecanoyloxy)hexadecanoic acid was attached at position 3'. The predominant absence of ester-bound 4'-phosphate and the presence of tetraacyl lipid A with fatty acids of 16 to 18 carbons in length differentiate H. pylori lipid A from that of other bacterial species and help explain the low endotoxic and biological activities of H. pylori LPS. PMID- 9335297 TI - Filamentous phage infection: required interactions with the TolA protein. AB - Infection of Escherichia coli by the filamentous phage f1 is initiated by binding of the phage to the tip of the F conjugative pilus via the gene III protein. Subsequent translocation of phage DNA requires the chromosomally encoded TolQ, TolR, and TolA proteins, after the pilus presumably has withdrawn, bringing the phage to the bacterial surface. Of these three proteins, TolA is proposed to span the periplasm, since it contains a long helical domain (domain II), which connects a cytoplasmic membrane anchor domain (domain I) to the carboxyl-terminal domain (domain III). By using a transducing phage, the requirement for TolA in an F+ strain was found to be absolute. The role of TolA domains II and III in the infective process was examined by analyzing the ability of various deletion mutants of tolA to facilitate infection. The C-terminal domain III was shown to be essential, whereas the polyglycine region separating domains I and II could be deleted with no effect. Deletion of helical domain II reduced the efficiency of infection, which could be restored to normal by retaining the C-terminal half of domain II. Soluble domain III, expressed in the periplasm but not in the cytoplasm or in the medium, interfered with infection of a tolA+ strain. The essential interaction of TolA domain III with phage via gene III protein appears to require interaction with a third component, either the pilus tip or a periplasmic entity. PMID- 9335298 TI - Contribution of different segments of the par region to stable maintenance of the broad-host-range plasmid RK2. AB - A 3.2-kb region of the broad-host-range plasmid RK2 has been shown to encode a highly efficient plasmid maintenance system that functions in a vector independent manner. This region, designated par, consists of two divergently arranged operons: parCBA and parDE. The 0.7-kb parDE operon promotes plasmid stability by a postsegregational killing mechanism that ensures that plasmid-free daughter cells do not survive after cell division. The 2.3-kb parCBA operon encodes a site-specific resolvase protein (ParA) and its multimer resolution site (res) and two proteins (ParB and ParC) whose functions are as yet unknown. It has been proposed that the parCBA operon encodes a plasmid partitioning system (M. Gerlitz, O. Hrabak, and H. Schwabb, J. Bacteriol. 172:6194-6203, 1990; R. C. Roberts, R. Burioni, and D. R. Helinski, J. Bacteriol. 172:6204-6216, 1990). To further define the role of this region in promoting the stable maintenance of plasmid RK2, the parCBA and parDE operons separately and the intact (parCBA/DE) par region (3.2 kb) were reintroduced into an RK2 plasmid deleted for par and assayed for plasmid stability in two Escherichia coli strains (MC1061K and MV10delta lac). The intact 3.2-kb region provided the highest degree of stability in the two strains tested. The ability of the parCBA or parDE region alone to promote stable maintenance in the E. coli strains was dependent on the particular strain and the growth temperature. Furthermore, the insertion of the ColE1 cer site into the RK2 plasmid deleted for the par region failed to stabilize the plasmid in the MC1061K strain, indicating that the multimer resolution activity encoded by parCBA is not by itself responsible for the stabilization activity observed for this operon. To examine the relative contributions of postsegregational cell killing and a possible partitioning function encoded by the intact 3.2-kb par region, stability assays were carried out with ParD provided in trans by a compatible (R6K) minireplicon to prevent postsegregational killing. In E. coli MV10delta lac, postsegregational killing appeared to be the predominant mechanism for stabilization since the presence of ParD substantially reduced the stability of plasmids carrying either the 3.2- or 0.7-kb region. However, in the case of E. coli MC1061K, the presence of ParD in trans did not result in a significant loss of stabilization by the 3.2-kb region, indicating that the putative partitioning function was largely responsible for RK2 maintenance. To examine the basis for the apparent differences in postsegregational killing between the two E. coli strains, transformation assays were carried out to determine the relative sensitivities of the strains to the ParE toxin protein. Consistent with the relatively small contribution of the postsegregational killing to plasmid stabilization in MC1061K, we found that this strain was substantially more resistant to killing by ParE in comparison to E. coli MV10delta lac. A transfer-deficient mutant of thepar-deleted plasmid was constructed for the stable maintenance studies. This plasmid was found to be lost from E. coli MV10delta lac at a rate three times greater than the rate for the transfer-proficient plasmid, suggesting that conjugation can also play a significant role in the maintenance of plasmid RK2. PMID- 9335299 TI - Clostridium perfringens epsilon-toxin acts on MDCK cells by forming a large membrane complex. AB - Epsilon-toxin is produced by Clostridium perfringens types B and D and is responsible for a rapidly fatal enterotoxemia in animals, which is characterized by edema in several organs due to an increase in blood vessel permeability. The Madin-Darby canine kidney (MDCK) cell line has been found to be susceptible to epsilon-toxin (D. W. Payne, E. D. Williamson, H. Havard, N. Modi, and J. Brown, FEMS Microbiol. Lett. 116:161-168, 1994). Here we present evidence that epsilon toxin cytotoxic activity is correlated with the formation of a large membrane complex (about 155 kDa) and efflux of intracellular K+ without entry of the toxin into the cytosol. Epsilon-toxin induced swelling, blebbing, and lysis of MDCK cells. Iodolabeled epsilon-toxin bound specifically to MDCK cell membranes at 4 and 37 labeled C and was associated with a large complex (about 155 kDa). The binding of epsilon-toxin to the cell surface was corroborated by immunofluorescence staining. The complex formed at 37 degrees C was more stable than that formed at 4 degrees C, since it was not dissociated by 5% sodium dodecyl sulfate and boiling. PMID- 9335301 TI - HrpA, a new ribosome-associated protein which appears in heat-stressed Mycobacterium bovis bacillus Calmette-Guerin. AB - A novel 18-kDa heat shock protein, HrpA, has been identified from Mycobacterium bovis BCG. HrpA was rapidly synthesized in membrane and ribosome fractions but not in the cytoplasmic fraction under heat shock stress. HrpA bound tightly to 70S ribosomes, mainly in 30S subunits. HrpA might be involved in the initiation step of translation at high temperature. PMID- 9335300 TI - Biochemical and genetic characterization of 2-carboxybenzaldehyde dehydrogenase, an enzyme involved in phenanthrene degradation by Nocardioides sp. strain KP7. AB - 2-Carboxybenzaldehyde dehydrogenase from the phenanthrene-degrading bacterium Nocardioides sp. strain KP7 was purified and characterized. The purified enzyme had a molecular mass of 53 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 205 kDa by gel filtration chromatography. Thus, the homotetramer of the 53-kDa subunit constituted an active enzyme. The apparent Km and kcat values of this enzyme for 2-carboxybenzaldehyde were 100 microM and 39 s(-1), respectively, and those for NAD+ were 83 microM and 32 s(-1), respectively. The structural gene for this enzyme was cloned and sequenced. The length of the gene was 1,455 bp. The nucleotide sequence of the 10,279 bp of DNA around the gene for 2-carboxybenzaldehyde dehydrogenase was also determined, and seven open reading frames were found in this DNA region. These were the genes for 1-hydroxy-2-naphthoate dioxygenase (phdI) and trans-2'-carboxybenzalpyruvate aldolase (phdJ), orf1, the gene for 2-carboxybenzaldehyde dehydrogenase (phdK), orf2/orf3, and orf4. The amino acid sequence of the orf1 product was similar to that of the aromatic hydrocarbon transporter gene (pcaK) in Pseudomonas putida PRS2000. The amino acid sequence of the orf4 product revealed a similarity to cytochrome P-450 proteins. The region between phdK and orf4 encoded orf2 and orf3 on different strands. The amino acid sequences of the orf2 and orf3 products exhibited no significant similarity to the reported sequences in protein databases. PMID- 9335302 TI - Characterization of a thermostable DNA photolyase from an extremely thermophilic bacterium, Thermus thermophilus HB27. AB - The photolyase gene from Thermus thermophilus was cloned and sequenced. The characteristic absorption and fluorescence spectra of the purified T. thermophilus photolyase suggested that the protein has flavin adenine dinucleotide as a chromophore. The second chromophore binding site was not conserved in T. thermophilus photolyase. The purified enzyme showed light dependent photoreactivation activity in vitro at 35 and 65 degrees C and was stable when subjected to heat and acidic pH. PMID- 9335303 TI - Evidence that rseC, a gene in the rpoE cluster, has a role in thiamine synthesis in Salmonella typhimurium. AB - In Salmonella typhimurium, the genetic loci and biochemical reactions necessary for the conversion of aminoimidazole ribotide (AIR) to the 4-amino-5 hydroxymethyl-2-methyl pyrimidine (HMP) moiety of thiamine remain unknown. Preliminary genetic analysis indicates that there may be more than one pathway responsible for the synthesis of HMP from AIR and that the function of these pathways depends on the availability of AIR, synthesized by the purine pathway or by the purF-independent alternative pyrimidine biosynthetic (APB) pathway (L. Petersen and D. Downs, J. Bacteriol. 178:5676-5682, 1996). An insertion in rseB, the third gene in the rpoE rseABC gene cluster at 57 min, prevented HMP synthesis in a purF mutant. Complementation analysis demonstrated that the HMP requirement of the purF rseB strain was due to polarity of the insertion in rseB on the downstream rseC gene. The role of RseC in thiamine synthesis was independent of rpoE. PMID- 9335304 TI - Genetic diversity in temperate bacteriophages of Streptococcus pyogenes: identification of a second attachment site for phages carrying the erythrogenic toxin A gene. AB - Bacteriophage T12, the prototypic bacteriophage of Streptococcus pyogenes carrying the erythrogenic toxin A gene (speA), integrates into the bacterial chromosome at a gene for a serine tRNA (W. M. McShan, Y.-F. Tang, and J. J. Ferretti, Mol. Microbiol. 23:719-728, 1997). This phage is a member of a group of related temperate phages, and we show here that not all speA-carrying phages in this group use the same attachment site for integration into the bacterial chromosome. Additionally, other phages in the group use the same serine tRNA gene attachment site as phage T12 and yet do not carry speA. The evidence suggests that recombination between phage genomes has been an important means of generating diversity and disseminating virulence-associated genes like speA. PMID- 9335305 TI - Two distinct models account for short and long deletions within sequence repeats in Escherichia coli. AB - In Escherichia coli, (GpC)n sequences cloned into plasmid DNA molecules are deletion-prone with the occurrence of both short (<2 bp) and long (>2 bp) deletion events. These repetitive tracts can be stabilized by interrupting the strict monotony of the repetition with a variant dinucleotide sequence. The stabilization of short deletion events that is mediated by the variant sequence is completely lost in E. coli mismatch repair-deficient strains. In contrast, this repair pathway has no influence on the frequency of occurrence of long deletion events, even in sequences containing the variant repeat. These results lead us to propose two distinct models to account for short and long deletions within repetitive sequences in E. coli. Furthermore, this study reveals that the deletions occur preferentially at the end of the repeat sequence that is distal with respect to the origin of replication. PMID- 9335306 TI - A mutational study of the site-specific cleavage of EC83, a multicopy single stranded DNA (msDNA): nucleotides at the msDNA stem are important for its cleavage. AB - Multicopy single-stranded DNA (msDNA) molecules consist of single-stranded DNA covalently linked to RNA. Such molecules are encoded by genetic elements called retrons. Unlike other retrons, retron EC83 isolated from Escherichia coli 161 produces RNA-free msDNA by site-specific cleavage of msDNA at 5'-TTGA/A-3', where the slash indicates the cleavage site. In order to investigate factors responsible for the msDNA cleavage, retron EC83 was treated with hydroxylamine and colonies were screened for cleavage-negative mutants. We isolated three mutants which were defective in msDNA cleavage and produced RNA-linked msDNA. They were all affected in msd, a gene for msDNA, with a base substitution at the bottom part of the msDNA stem. In contrast, base substitution at and around the cleavage site has no marked effect on msDNA synthesis or its cleavage. From these results, we concluded that the nucleotides at the bottom of the msDNA stem, but not the nucleotides at the cleavage site, play a major role in the recognition and cleavage of msDNA EC83. PMID- 9335307 TI - The Serratia marcescens NucE protein functions as a holin in Escherichia coli. AB - The recently discovered nucC locus of Serratia marcescens encodes the cryptic prophage genes nucE, nucD, and nucC. NucC is required for expression of the S. marcescens nuclease and functions as a transcriptional activator of the nuclease gene, nucA. NucE and NucD are dispensable for nuclease expression but were proposed to allow for secretion of the nuclease by Escherichia coli. Here, we show (i) that the NucE protein is membrane bound, (ii) that it can complement the lambda S holin, (iii) that it can be triggered by potassium cyanide, (iv) that it is detrimental to cell viability, and (v) that the concomitant expression of nucE and nucD results in cell lysis. Apparently NucE and NucD function as a holin and an endolysin, respectively. This suggests that their roles in nuclease secretion by E. coli are indirect, possibly through directed cell lysis. PMID- 9335309 TI - Temporal regulation of sigD from Bacillus subtilis depends on a minor promoter in front of the gene. AB - I investigated the transcriptional regulation of sigmaD synthesis. sigD is part of the fla/che operon, but the gene is also preceded by a promoter of its own. fla/che-dependent transcription is severely reduced in sigD-negative strains. Activity of the promoter in front of sigD is strictly temporally regulated. Deletion of this promoter results in a reduced and delayed activation of transcription of the fla/che operon. PMID- 9335308 TI - Transcriptional regulation of the cydDC operon, encoding a heterodimeric ABC transporter required for assembly of cytochromes c and bd in Escherichia coli K 12: regulation by oxygen and alternative electron acceptors. AB - The expression of the cydDC operon was investigated by using a chromosomal phi(cydD-lacZ) transcriptional fusion and primer extension analysis. A single transcriptional start site was found for cydD located 68 bp upstream of the translational start site, and Northern blot analysis confirmed that cydDC is transcribed as a polycistronic message independently of the upstream gene trxB. cydDC was highly expressed under aerobic growth conditions and during anaerobic growth with alternative electron acceptors. Aerobic expression was independent of ArcA and Fnr, but induction of cydDC by nitrate and nitrite was dependent on NarL and Fnr. PMID- 9335311 TI - Clues to immune function and oncogenesis provided by events that activate the cell cycle machinery in normal human T cells. AB - A common feature seen in states of decreased immune competence or immunosuppression and in diseases of the blood, such as lymphohematopoietic cancers, is the disruption of the normal pathways of cell cycle control. In lymphocytes a series of nonlinear biochemical cascades leads to cellular proliferation and also controls the production of cytokines that provide immunologic help (i.e., aid in B and T cell proliferation, maturation, and differentiation). These two distinct outcomes can be dissociated, as stimuli that incite production of cytokines need not lead to cell division, and conversely, exogenously provided cytokines may promote lymphocyte proliferation. The signals that induce production of cytokines, particularly interleukin-2, have been extensively characterized. It also is known that the fidelity of cell cycle progression is dependent on a regulatory network whose key components include cyclin-dependent kinases and cyclins. This review describes the current state of knowledge linking the antigen receptor response pathways and the activation of the cell cycle machinery in T cells. PMID- 9335310 TI - Microorganisms and their interaction with the immune system. AB - Microorganisms interact with the immune system in multiple ways. In an interaction between a microorganism and its host, the defense of the host does not go unchallenged. Microorganisms have for decades been suspected of possessing the capabilities of hiding from and escaping the consequences of immune surveillance. Escape mechanisms like antigenic variation, latency, and genomic integration can best be described as passive mechanisms for avoiding interaction with the host immune system, to differentiate them from the more engaging and host-directed active mechanisms of interaction. Studies of the mechanism of direct entry of viruses (HIV, measles, and enteroviruses), bacteria (streptococci and staphylococci), and parasites (Leishmania and plasmodium) into immune cells like CD4+ T cells or macrophages, as reported very recently, indicate an even more aggressive mode of interaction. This aggressive mechanism of interaction with the components of the host immune system allows the microbe not only to block the normal function of immune components on the surface of immune cells from functioning, but also to obliterate a vital immune function, cellular immunity, causing immunosuppression, e.g. the depletion of CD4+ T cells due to the entry and replication of HIV. Collectively, microorganisms have evolved various mechanisms by which they can actively block almost any cellular, humoral, or systemic immune response. One general feature of the proteins that assist microorganism to immunomodulate and actively evade host defense is their structural and therefore functional similarity to the host proteins, which they effectively mimic. Understanding the different mechanisms by which microorganisms interact with the immune system can impact the design of live vaccines as well as the development of novel therapeutic immunomodulators that can provide medicine with powerful new tools to manage immune disorders, allograft rejection, remote multiple organ failure resulting from trauma, autoimmune diseases, etc. PMID- 9335312 TI - Oral administration of unsaturated fatty acids: effects on human peripheral blood T lymphocyte proliferation. AB - Oils enriched in certain polyunsaturated fatty acids suppress joint pain and swelling in rheumatoid arthritis (RA) patients. Because T lymphocyte activation is important to propagation of joint tissue injury in patients with RA, we examined the effects of fatty acids administered by mouth in vivo on proliferation of human lymphocytes activated through the T cell receptor complex. T cell proliferation was reduced after oral administration of 2.4 g gammalinolenic acid in capsules of borage seed oil. Oral administration of oils enriched in linoleic acid, the parent n-6 fatty acid, and alpha linolenic acid, the parent n-3 fatty acid, did not influence growth of stimulated cells. Fatty acid analyses indicated that suppression of lymphocyte proliferation after gammalinolenic acid administration was associated with increased plasma and peripheral blood mononuclear cell concentrations of gammalinolenic acid and dihomogammalinolenic acid. PMID- 9335313 TI - Attenuation of monosodium urate crystal-induced arthritis in rabbits by a neutralizing antibody against interleukin-8. AB - Accumulating evidence implicates interleukin-8 (IL-8) as an essential mediator in neutrophil-mediated acute inflammation. Neutrophils have also been shown to have a crucial role in the pathogenesis of acute gouty arthritis. Thus, we investigate the pathophysiological role of IL-8 in an experimental model of acute gout, monosodium urate (MSU) crystal-induced arthritis in rabbits. The injection of MSU crystals into knee joints caused a marked swelling of joints. Concomitantly, the infiltration ofleukocytes, mostly neutrophils, was observed in synovial membrane and synovial fluids. The injection of MSU crystals also induced an elevation in synovial fluid IL-8 levels preceding neutrophil infiltration into synovial fluids, without an accompanying increase in plasma IL-8 levels. Immunoreactive IL 8 protein was detected in synovial lining cells at 12-24 h after the injection. IL-8 protein was also observed in infiltrated leukocytes in synovium as early as 3-24 h after the injection. Finally, the intraarticular injection of a neutralizing anti-IL-8 antibody significantly attenuated the crystal-induced joint swelling that occurred at 12 h, and neutrophil infiltration into arthritic joints at 12 and 24 h after the induction. These results provide evidence on the pathogenic roles of locally produced IL-8 in MSU crystal-induced gouty arthritis. PMID- 9335315 TI - The role of shear forces in ischemia/reperfusion-induced neutrophil rolling and adhesion. AB - Intravital microscopy was used to examine the role of reduced shear forces on neutrophil-endothelium interactions in single 25-40 microm diameter vessels in the feline postischemic mesenteric microvasculature. Neutrophil rolling, neutrophil adhesion, and microvascular permeability alterations were determined in vessels exposed to ischemia/reperfusion in untreated animals and in cats that received feline plasma directly into the superior mesenteric artery to increase intestinal blood flow and venular shear rates. Ischemia, followed by reperfusion, caused a profound increase in venular shear forces during the initial hyperemic response, which then decreased to below control values by 10 min of reperfusion and to less than 50% of control by 60 min of reperfusion. Associated with the decrease in blood flow was a profound increase in neutrophil rolling and neutrophil adhesion and an increase in microvascular permeability. Also, leukocyte rolling velocity decreased dramatically to 10-20% of control values during the first 10 min of reperfusion. Infusion of autologous plasma into the intestinal vasculature to maintain the shear forces at control levels during reperfusion did not affect the flux of rolling neutrophils. The rolling velocity of the neutrophils was not increased despite dramatically improved shear rates. Improved shear rates did reduce the number of adherent cells, resulting in less vascular dysfunction. The reduction in shear forces through inflamed microvessels does not contribute to the increased leukocyte rolling flux but is an essential, permissive component for neutrophil adhesion and subsequent vascular dysfunction in postischemic microvasculature. PMID- 9335314 TI - IL-12-induced tumor regression correlates with in situ activity of IFN-gamma produced by tumor-infiltrating cells and its secondary induction of anti-tumor pathways. AB - Administration of recombinant interleukin-12 (rIL-12) into CSA1M fibrosarcoma bearing mice results in complete regression of growing tumors. This tumor regression is associated with massive lymphoid cell infiltration to tumor sites and is completely blocked by injection of anti-interferon-gamma (IFN-gamma) monoclonal antibody (mAb). We investigated whether anti-IFN-gamma mAb exerts its suppressive effect on tumor regression by blocking the IL-12-induced lymphoid cell migration to tumor sites or by inhibiting the secondary effects of IFN-gamma produced by infiltrating cells. Injection of anti-IFN-gamma mAb to CSA1M-bearing mice before IL-12 treatment prevented the induction of tumor regression, whereas this treatment affected only marginally the infiltration of lymphoid cells to tumor masses. In accordance with this, IFN-gamma mRNA was expressed inside tumor masses by infiltrating cells after IL-12 therapy irrespective of whether anti-IFN gamma mAb was injected. However, anti-IFN-gamma mAb treatment almost completely abrogated the in situ expression of inducible nitric oxide synthase (iNOS) as well as IFN-inducible protein-10 (IP-10) genes as examples of IFN-gamma-inducible genes. Immunohistochemical analyses also revealed that the expression of iNOS protein was completely inhibited by anti-IFN-gamma injection. These results suggest that the implementation of in situ IFN-gamma activity and its secondary induction of anti-tumor pathways such as iNOS and IP-10 expression are important processes in the IL-12-induced tumor regression. PMID- 9335316 TI - Adenosine A3 receptor stimulation inhibits migration of human eosinophils. AB - Activation of adenosine A3 receptors (A3-R) produced a dose-dependent reduction in the chemotaxis of human eosinophils to platelet-activating factor (PAF), RANTES, and leukotriene B4 (LTB4) to a maximum of 58, 48, and 52%, respectively (P < 0.02). This effect was completely reversed by selective A3-R antagonists. In contrast, activation of A1 or A2a-R did not affect PAF-induced eosinophil chemotaxis. PAF up-regulated the expression of CDllb/CD18, down-regulated L selectin, and also increased F-actin assembly in eosinophils. The expression of these activation markers was not influenced by A3-R, A2a, or A1-R stimulation. Activation of A3-R may play an important role in inflammation by inhibiting eosinophil migration. PMID- 9335317 TI - Humanized mAb H22 binds the human high affinity Fc receptor for IgG (FcgammaRI), blocks phagocytosis, and modulates receptor expression. AB - About 10-15% of patients with immune thrombocytopenic purpura (ITP) cannot be controlled by corticosteroid therapy and splenectomy. For these patients treatment with high-dose IVIgG induces partial or complete responses. The clinical benefits of IVIgG could be due to blockade of Fc receptors for IgG (FcgammaR), because several model systems clearly show that functional FcgammaR are essential for establishment of ITP and related diseases. However, the specific contributions of the three individual classes of FcgammaR remain to be more completely defined. Recently monoclonal antibody (mAb) H22, which recognizes an epitope on FcgammaRI (CD64) outside the ligand binding domain, was humanized by grafting its complementarity determining regions onto human IgG1 constant domains. Because FcgammaRI has a high affinity for human IgG1 antibodies, we predicted mAb H22 would also bind to FcgammaRI through its Fc domain and block FcgammaRI-mediated phagocytosis. These studies demonstrate that mAb H22 blocked phagocytosis of opsonized red blood cells 1000 times more effectively than an irrelevant IgG. Moreover, cross-linking FcgammaRI with mAb H22 rapidly down modulated FcgammaRI expression on monocytes without affecting other surface antigens. We conclude that because mAb H22 is a humanized mAb that blocks the FcgammaRI ligand binding domain and down-modulates FcgammaRI expression, it is a particularly good candidate for evaluating the role of FcgammaRI in patients with ITP. PMID- 9335318 TI - Characterization of a culture-derived CD15+CD11b- promyelocytic population from CD34+ peripheral blood cells. AB - Selected CD34+ cells from mobilized apheresis products were cultured in serum free or serum-containing media supplemented with granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and stem cell factor (SCF; c-kit ligand). We examined the emergence of a CD15+CD11b- population, which appeared morphologically to be promyelocytes. This CD15+CD11b- population can be further expanded in culture into morphologically mature granulocytes. In an attempt to characterize this culture-derived CD15+CD11b- promyelocytic population, single cells were clone sorted into wells of a Terasaki plate containing various growth factors. We compared the growth factor requirements and kinetics of this apheresis culture derived CD15+CD11b- population to the CD15+CD11b- population from fresh bone marrow samples. Our studies indicate that the CD15+CD11b- promyelocytic population from bone marrow and blood are equivalent in their ability to proliferate and in their requirements for growth factors. The CD15+CD11b- population in vitro shows a high proliferative capacity when compared with the other CD15/CD11b populations (CD15-CD11b-, CD15+CD11b+, CD15-CD11b+). Thus, we can manipulate CD34+ cells in vitro to proliferate and differentiate toward a mature neutrophil lineage. The CD15+CD11b- promyelocytic population derived from this culture may represent the most effective cultured cell population for therapeutic reduction of neutropenia in vivo based on both its stage of differentiation and its proliferative potential. PMID- 9335319 TI - Monocytes incubated with surfactant: a model for human alveolar macrophages? AB - Monocytes migrate to the lungs and enter the alveoli where they come into contact with surfactant and differentiate into alveolar macrophages. This study focused on the question of the extent to which monocytes and monocyte-derived macrophages (MDM) incubated with surfactant resemble alveolar macrophages. Surfactant incubated monocytes shared with alveolar macrophages the intracellular presence of surfactant, efficient phagocytosis of opsonized Staphylococcus aureus, and poor intracellular killing of ingested bacteria. The suppressive effect of surfactant on bactericidal activities of monocytes could not be attributed to either the surfactant lipid fraction or surfactant protein A. Monocytes incubated with surfactant differed from alveolar macrophages with respect to expression of various Fc and complement receptors involved in intracellular killing of bacteria. Surfactant-incubated monocytes produced significantly more H2O2 upon stimulation with phorbol ester than alveolar macrophages, but significantly less than control monocytes. Together, monocytes and MDM incubated with surfactant, although similar to alveolar macrophages in some aspects, are not an adequate model for alveolar macrophages. Most likely, factors other than surfactant in the microenvironment of the alveoli, such as oxygen tension, play a role in the differentiation of monocytes to alveolar macrophages as well. PMID- 9335321 TI - Enhanced mobilization of hematopoietic progenitor cells by mouse MIP-2 and granulocyte colony-stimulating factor in mice. AB - Human interleukin-8 (IL-8) induces a rapid mobilization of hematopoietic progenitor cells (HPCs) into peripheral blood in mice and primates. Because an exact homologue of human IL-8 does not exist in mice, a chemokine, macrophage inflammatory protein-2 (MIP-2), is supposed to substitute the function of IL-8 in mice. Hence, we investigated the capacity of mouse MIP-2 to induce HPC mobilization in mice. Mouse MIP-2 induced, in either untreated or splenectomized mice, a rapid HPC mobilization, as evidenced by increased numbers of colony forming unit granulocyte-macrophage (CFU-GM) and lineage marker (Lin)-c-kit+Sca 1+ cells. Although release of elastase from activated neutrophils was proposed to be involved in IL-8-induced HPC mobilization, mouse MIP-2 induced similar levels of HPC mobilization in elastase-deficient beige mice at a similar level to normal ones. This HPC mobilization by MIP-2 was markedly enhanced by pretreatment with granulocyte-colony-stimulating factor (G-CSF). Moreover, the combination of G-CSF and MIP-2 apparently down-regulated L-selectin expression on Lin-Sca-1+ cells in peripheral blood as well as bone marrow. Thus, MIP-2 may down-modulate the interaction between HPCs and bone marrow stroma by reducing L-selectin expression on HPCs and cause rapid mobilization of HPCs into peripheral blood. PMID- 9335320 TI - In vitro expansion of CD13+CD33+ dendritic cell precursors from multipotent progenitors is regulated by a discrete fas-mediated apoptotic schedule. AB - We provide new information on how apoptosis regulates the expansion and survival of dendritic cell (DC) elements during in vitro hematopoiesis. Functionally distinct apoptotic schedules were associated with different phases of DC development when multipotent CD34+ progenitor cells were treated with GM-CSF + TNF +/- SCF (c-kit ligand). During early phases of growth, unselected progenitors underwent apoptosis. During intermediate stages, high levels of apoptosis resulted in the preferential selection of DC precursors, as revealed by the massive expansion of DR+CD33+CD13+ cells. Late apoptosis was associated with the death of mature DCs. Apoptotic events surrounding the earlier periods were related to the exogenous addition of TNF-alpha and appeared to be mediated by fas. In contrast, those events associated with terminally differentiated DCs were fas independent because there was no correlation between fas expression and cell death. The bcl-2 protein family appeared to confer resistance to apoptotic death, as revealed by the high levels of bcl-2 and bclxL during peak DC development and in long-term DC cultures. We demonstrate that activation of distinct apoptotic programs regulates DC development and homeostasis. Although suppression of apoptosis may prolong the survival of late DC elements, an earlier apoptotic schedule appears to be required for the selective expansion of DC elements from multipotent progenitors. Our data also provides insight into the mechanism(s) of myeloid lineage selection by cytokines such as TNF-alpha, which may promote both cell death and survival. PMID- 9335322 TI - Up-regulation of Tie gene expression by leukemia inhibitory factor and steel factor in CD34+ cells from human umbilical cord blood. AB - Tie, a new receptor tyrosine kinase, is expressed in vascular endothelial and hematopoietic cells. To determine whether Tie might be involved in early hematopoiesis, we asked whether the Tie gene is expressed in normal human hematopoietic stem/progenitor cells and if the expression of this gene could be regulated. Using a single-cell reverse-transcriptase polymerase chain reaction (RT-PCR) assay to study expression of the Tie gene in the subset of human umbilical cord blood (UCB) CD34+++ primitive stem/ progenitor cells with extensive replating capacity, we demonstrated at the single isolated cell level that Tie was expressed in these cells. The expression of Tie gene in CD34+ cells was at a low level but was enhanced up to two- to fourfold by steel factor (SLF) or leukemia inhibitory factor (LIF), two cytokines that regulate production of stem/progenitor cells, as assessed using competitive PCR and semi-quantitative RT PCR assays. The fold increases were observed as early as 2 h for SLF and 4 h for LIF and remained elevated for 24 h. These results demonstrate modulation of gene regulation in the rare populations of CD34+ cells and suggest the possibility that Tie may play a role in the proliferation and differentiation of immature hematopoietic cells. PMID- 9335323 TI - Ultraviolet irradiation accelerates apoptosis in human polymorphonuclear leukocytes: protection by LPS and GM-CSF. AB - Polymorphonuclear leukocytes (PMN) play a central role in host response to injury and infection. Understanding factors that regulate PMN survival may therefore have a major influence on the development of novel treatment strategies for controlling life-threatening infections, as well as local and systemic inflammatory responses. Unfortunately, the presently utilized in vitro culture model of PMN apoptosis makes the examination of early biochemical events surrounding PMN apoptosis very difficult. This study demonstrates that a short course of UV irradiation (15 min) can be used to induce rapid progression of PMN through the apoptotic process with 70-90% of PMN displaying features of apoptosis by 4 h after UV exposure. Bacterial lipopolysaccharide and granulocyte-macrophage colony-stimulating factor, which are known to prolong PMN survival during in vitro culture, also protected PMN from UV-accelerated apoptosis. The UV accelerated model of PMN apoptosis provides another valuable tool for the investigation of early signaling pathways associated with inducing or delaying PMN apoptosis. PMID- 9335325 TI - Relationship of ligand-receptor dynamics to actin polymerization in RBL-2H3 cells transfected with the human formyl peptide receptor. AB - The human formyl peptide receptor (FPR) expressed in RBL-2H3 transfectants (RBL[FPR]) behaves qualitatively like the FPR expressed by neutrophils except that it causes sustained F-actin accumulation and cell shape change responses on formyl peptide stimulation. These sustained responses were not accounted for by changes in the transfected receptor's ability to interact with ligand or by receptor density. Signal transduction pathways of transfected and neutrophil FPRs are apparently similar. In transfected cells, dissociation of ligand is sensitive to guanine nucleotide, the G protein is pertussis toxin-sensitive, FPR and G protein appear to be precoupled, the F-actin response is stimulated with the same dose-response profile as in neutrophils, and the F-actin accumulation response is directly regulated by the FPR, even long after initial stimulation. Potentially significant differences between neutrophil and transfected FPR were found when receptor processing was measured. In neutrophils, practically 100% of the FPR is converted to forms that dissociate slowly from ligand and are inactive in signal transduction within 2 min of ligand stimulation. By contrast, 20% or more of transfected FPR remains rapidly dissociating even 5 min after stimulation. Although 80% of neutrophil FPR is internalized by 5 min after stimulation, transfected FPR appears to plateau at 50-60% internalized. Because actin responses in neutrophils are regulated by a small number of active receptors, the inefficiency of receptor inactivation in RBL(FPR) transfectants may account for the prolonged F-actin accumulation response. PMID- 9335324 TI - Increased monocyte TNF-alpha message stability contributes to trauma patients' increased TNF production. AB - Post-trauma elevation of tumor necrosis factor alpha (TNF-alpha) appears to be critical in mediating many symptoms of systemic inflammatory response syndrome (SIRS), resulting in late mortality. Although increased monocyte (mphi) TNF-alpha production plays a pivotal role in this TNF-alpha elevation, the molecular mechanisms leading to increased mphi TNF-alpha production have yet to be elucidated. We demonstrate that, although TNF-alpha mRNA levels are increased in all trauma patients' mphi, which produce elevated levels of TNF-alpha protein, in the majority of patients, these increased TNF-alpha mRNA levels are under normal transcriptional and posttranscriptional control. Consequently, the increased TNF alpha production by these patients' mphi is probably due to preactivation of these mphi by trauma-released mediators. However, a small minority of patients, whose mortality rate was 57%, produce TNF-alpha of primarily the membrane associated type. The mphi TNF-alpha mRNA accumulation of these patients in response to in vitro stimulation is significantly augmented. All of these patients experienced SIRS. In this subset of patients' mphi, TNF-alpha mRNA stability was aberrantly increased. Such an increase in TNF-alpha mRNA stability could lead to devastatingly prolonged production of TNF-alpha protein. This demonstration of increased TNF-alpha mRNA stability in post-trauma mphi represents a novel correlation of elevated membrane-associated TNF-alpha protein, increased mortality, and a mechanism for this occurrence. PMID- 9335326 TI - Suppression of TNF-alpha gene expression by hemin: implications for the role of iron homeostasis in host inflammatory responses. AB - Tumor necrosis factor alpha (TNF-alpha) has multiple effects on iron homeostasis and erythropoiesis and has been implicated in the pathogenesis of the anemia of inflammation. We postulated that intracellular iron in turn may regulate the expression of TNF-alpha. In the human monocytic leukemia cell line, THP-1, low basal TNF-alpha message levels were stimulated (sevenfold) when serum was excluded from the culture medium. Addition of hemin completely suppressed TNF alpha expression. Similarly, hemin suppressed lipopolysaccharide-induced expression of TNF-alpha. Sn-protoporphyrin, an inhibitor of heme oxygenase (which releases iron from hemin), prevented hemin-induced suppression of TNF-alpha expression. Conversely, the intracellular iron chelator, desferrioxamine, stimulated TNF-alpha expression. Thus, the expression of TNF-alpha, itself a physiological regulator of iron homeostasis, appears to be controlled by intracellular levels of iron. PMID- 9335327 TI - Life and cancer without telomerase. PMID- 9335328 TI - Condensins, cohesins, and chromosome architecture: how to make and break a mitotic chromosome. PMID- 9335329 TI - Transient Xist-ence. PMID- 9335330 TI - Multiple TATA-binding factors come back into style. PMID- 9335331 TI - La dolce vita: a molecular feast in plant-pathogen interactions. PMID- 9335332 TI - Telomere shortening and tumor formation by mouse cells lacking telomerase RNA. AB - To examine the role of telomerase in normal and neoplastic growth, the telomerase RNA component (mTR) was deleted from the mouse germline. mTR-/- mice lacked detectable telomerase activity yet were viable for the six generations analyzed. Telomerase-deficient cells could be immortalized in culture, transformed by viral oncogenes, and generated tumors in nude mice following transformation. Telomeres were shown to shorten at a rate of 4.8+/-2.4 kb per mTR-/- generation. Cells from the fourth mTR-/- generation onward possessed chromosome ends lacking detectable telomere repeats, aneuploidy, and chromosomal abnormalities, including end-to-end fusions. These results indicate that telomerase is essential for telomere length maintenance but is not required for establishment of cell lines, oncogenic transformation, or tumor formation in mice. PMID- 9335333 TI - Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. AB - Cohesion between sister chromatids opposes the splitting force exerted by microtubules, and loss of this cohesion is responsible for the subsequent separation of sister chromatids during anaphase. We describe three chromosmal proteins that prevent premature separation of sister chromatids in yeast. Two, Smc1p and Smc3p, are members of the SMC family, which are putative ATPases with coiled-coil domains. A third protein, which we call Scc1p, binds to chromosomes during S phase, dissociates from them at the metaphase-to-anaphase transition, and is degraded by the anaphase promoting complex. Association of Scc1p with chromatin depends on Smc1p. Proteins homologous to Scc1p exist in a variety of eukaryotic organisms including humans. A common cohesion apparatus might be used by all eukaryotic cells during both mitosis and meiosis. PMID- 9335335 TI - Components and dynamics of DNA replication complexes in S. cerevisiae: redistribution of MCM proteins and Cdc45p during S phase. AB - In S. cerevisiae, the chromatin structure of DNA replication origins changes as cells become competent for DNA replication, suggesting that G1 phase-specific association of replication factors with origin DNA regulates entry into S phase. We demonstrate that ORC, Cdc45p, and MCM proteins are components of prereplication complexes (pre-RC). The MCM-origin association is dependent upon ORC and Cdc6p. During S phase, MCM proteins and Cdc45p dissociate from origin DNA and associate with nonorigin DNA with similar kinetics as DNA Polymerase epsilon, which is present at DNA replication forks. Our results identify protein components of the pre-RC and a novel replication complex appearing at the G1/S transition (the RC), and suggest that after initiation MCM proteins and Cdc45p move with eukaryotic replication forks. PMID- 9335334 TI - A direct link between sister chromatid cohesion and chromosome condensation revealed through the analysis of MCD1 in S. cerevisiae. AB - The S. cerevisiae MCD1 (mitotic chromosome determinant) gene was identified in genetic screens for genes important for chromosome structure. MCD1 is essential for viability and homologs are found from yeast to humans. Analysis of the mcd1 mutant and cell cycle-dependent expression pattern of Mcd1p suggest that this protein functions in chromosome morphogenesis from S phase through mitosis. The mcd1 mutant is defective in sister chromatid cohesion and chromosome condensation. The physical association between Mcd1p and Smc1p, one of the SMC family of chromosomal proteins, further suggests that Mcd1p functions directly on chromosomes. These data implicate Mcd1p as a nexus between cohesion and condensation. We present a model for mitotic chromosome structure that incorporates this previously unsuspected link. PMID- 9335336 TI - Transcription properties of a cell type-specific TATA-binding protein, TRF. AB - Eukaryotic cells are thought to contain a single TATA-binding protein (TBP) that directs transcription by cellular RNA polymerases. Here we report a cell type specific TBP-related factor (TRF) that can form a stable TRF/IIA/IIB TATA DNA complex and substitute for TBP in directing RNA polymerase II transcription in vitro. Transfection studies reveal that TRF can differentially mediate activation by some enhancer proteins but not others. Like TBP, TRF forms a stable complex containing multiple novel subunits, nTAFs. Antibody staining of embryos and polytene chromosomes reveals cell type-specific expression and gene-selective properties consistent with the shaker/male sterile phenotype of trf mutants. These findings suggest TRF is a homolog of TBP that functions to direct tissue- and gene-specific transcription. PMID- 9335337 TI - Crystal structure of a Hedgehog autoprocessing domain: homology between Hedgehog and self-splicing proteins. AB - The approximately 25 kDa carboxy-terminal domain of Drosophila Hedgehog protein (Hh-C) possesses an autoprocessing activity that results in an intramolecular cleavage of full-length Hedgehog protein and covalent attachment of a cholesterol moiety to the newly generated amino-terminal fragment. We have identified a 17 kDa fragment of Hh-C (Hh-C17) active in the initiation of autoprocessing and report here its crystal structure. The Hh-C17 structure comprises two homologous subdomains that appear to have arisen from tandem duplication of a primordial gene. Residues in the Hh-C17 active site have been identified, and their role in Hedgehog autoprocessing probed by site-directed mutagenesis. Aspects of sequence, structure, and reaction mechanism are conserved between Hh-C17 and the self splicing regions of inteins, permitting reconstruction of a plausible evolutionary history of Hh-C and the inteins. PMID- 9335338 TI - Stabilization of Xist RNA mediates initiation of X chromosome inactivation. AB - The onset of X inactivation is preceded by a marked increase in the level of Xist RNA. Here we demonstrate that increased stability of Xist RNA is the primary determinant of developmental up-regulation. Unstable transcript is produced by both alleles in XX ES cells and in XX embryos prior to the onset of random X inactivation. Following differentiation, transcription of unstable RNA from the active X chromosome allele continues for a period following stabilization and accumulation of transcript on the inactive X allele. We discuss the implications of these findings in terms of models for the initiation of random and imprinted X inactivation. PMID- 9335339 TI - The AP-3 adaptor complex is essential for cargo-selective transport to the yeast vacuole. AB - Three distinct adaptor protein (AP) complexes involved in protein trafficking have been identified. AP-1 and AP-2 mediate protein sorting at the trans-Golgi network and plasma membrane, respectively, whereas the function of AP-3 has not been defined. A screen for factors specifically involved in transport of alkaline phosphatase (ALP) from the Golgi to the vacuole/lysosome has identified Ap16p and Ap15p of the yeast AP-3 complex. Deletion of each of the four AP-3 subunits results in selective mislocalization of ALP and the vacuolar t-SNARE, Vam3p (but not CPS and CPY), while deletion of AP-1 and AP-2 subunits has no effect on vacuolar protein delivery. This study, therefore, provides evidence that the AP-3 complex functions in cargo-selective protein transport from the Golgi to the vacuole/lysosome. PMID- 9335340 TI - Reactivation of latent human cytomegalovirus by allogeneic stimulation of blood cells from healthy donors. AB - Reactivation of human cytomegalovirus (HCMV) results in severe disease in AIDS patients and immunocompromised patients receiving blood transfusions or organ or bone marrow grafts. Although the site of HCMV latency is unknown, blood cells have been implicated as a viral reservoir. In this study, we demonstrate HCMV reactivation in vitro from seven consecutive healthy donors through allogeneic stimulation of peripheral blood mononuclear cells (PBMCs). HCMV replication was detected at 17 days poststimulation, and virus was recovered after long-term culture from a macrophage expressing dendritic cell markers. Thus, these observations demonstrate that PBMCs harbor latent HCMV, which reactivates in a myeloid lineage cell upon allogeneic stimulation. PMID- 9335341 TI - A role for the roof plate and its resident TGFbeta-related proteins in neuronal patterning in the dorsal spinal cord. AB - Distinct neuronal cell types are generated at characteristic times and positions in the dorsal horn of the spinal cord. We provide evidence that the identity and pattern of generation of dorsal neurons depend initially on BMP-mediated signals that derive from the epidermal ectoderm and induce dorsal midline cells of the roof plate. Roof plate cells provide a secondary source of TGFbeta-related signals that are required for the generation of distinct classes of dorsal interneurons. These inductive interactions involve both qualitative and quantitative differences in signaling by TGFbeta-related factors and temporal changes in the response of neural progenitor cells. PMID- 9335342 TI - Sodium/hydrogen exchanger gene defect in slow-wave epilepsy mutant mice. AB - The "housekeeping" sodium/hydrogen exchanger, NHE1, mediates the electroneutral 1:1 exchange of Na+ and H+ across the plasma membrane. NHE1 is ubiquitous and is studied extensively for regulation of pHi, cell volume, and response to growth factors. We describe a spontaneous mouse mutant, slow-wave epilepsy, (swe), with a neurological syndrome including ataxia and a unique epilepsy phenotype consisting of 3/sec absence and tonic-clonic seizures. swe was fine-mapped on Chromosome 4 and identified as a null allele of Nhe1. Mutants show selective neuronal death in the cerebellum and brainstem but otherwise are healthy. This first example of a disease-causing mutation in an Nhe gene provides a new tool for studying the delicate balance of neuroexcitability and cell survival within the CNS. PMID- 9335343 TI - Bone formation by cells from femurs cultured among three-dimensionally arranged hydroxyapatite granules. AB - In vitro bone formation by cells derived from adult rabbit femurs was investigated on or in several substrates with small porous hydroxyapatite granules (HAGs). When the bone fragments were cultured in HAG-packed glass tubes, which were inclined (5 degrees -30 degrees ) and rotated 90 degrees per day after one week of culture, thin lamellar tissues were newly formed in narrow spaces among the HAGs. By 11 days of culture, these tissues had been mineralized except for their periphery and had well developed collagen bundles and several monolayer cells. Some cells resided in bone lacuna-like spaces. By contrast, mineralization was negligible in 6-week cultures on two-dimensional glass and polystyrene plates with or without two-dimensionally arranged HAGs on their surfaces and in three dimensional collagen gels with or without HAGs in spite of active cell proliferation. These results suggest that osteogenesis is accelerated in a specific three-dimensional constitution of extracellular matrix and/or under the effects of mechanical forces for the new tissue and that bioactive HAGs offer favorable three-dimensional spaces for osteogenic tissue formation. PMID- 9335344 TI - The detachment strength and morphology of bone cells contacting materials modified with a peptide sequence found within bone sialoprotein. AB - Adhesion, spreading, and focal contact formation of primary bone-derived cells on quartz surfaces grafted with a 15 amino acid peptide that contained a -RGD-(-Arg Gly-Asp-) sequence unique to bone sialoprotein was investigated. The peptide surfaces were fabricated by using a heterbifunctional crosslinker, sulfosuccinimidyal 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, to link the peptide to amine functionalized quartz surfaces. Contact angle measurements, spectroscopic ellipsometry, and X-ray photoelectron spectroscopy were used to confirm the chemistry and thickness of the overlayers. A radial flow apparatus was used to characterize cell detachment from peptide-grafted surfaces. After 20 min of cell incubation, the strength of cell adhesion was significantly (p < 0.05) higher on the -RGD- compared to -RGE- (control) surfaces. Furthermore, the mean area of cells contacting the -RGD- was significantly (p < 0.05) higher than RGE- surfaces. Vinculin staining showed formation of small focal contact patches on the periphery of bone cells incubated for 2 h on the -RGD- surfaces; however, few or no focal contacts were formed by cells seeded on the -RGE-grafted surfaces. The methods of peptide immobilization utilized in this study can be applied to implants, biosensors, and diagnostic devices that require specificity in cell adhesion. PMID- 9335345 TI - The protective effect of zinc on rosin and resin acid toxicity in human polymorphonuclear leukocytes and human gingival fibroblasts in vitro. AB - Combinations of rosin and zinc are used in dentistry as components of periodontal dressings and cements and as root canal sealers. The composition and properties of rosins differ largely depending on source and refinement processes. Rosin (colophony) is composed of approximately 70% resin acids. In order to study the toxic effects of different natural rosins and purified resin acids and the detoxifying effects of zinc, these compounds were analyzed and tested on human polymorphonuclear leukocytes (PMN cells) and human gingival fibroblasts using the radiochromium release method. The rosins and the pure resin acids showed a strong dose-related cytotoxicity, which was inhibited by increased zinc concentrations. The purified resin acids (isopimaric, levopimaric, and neoabietic acid) were more toxic than the natural rosins. The contents of these resin acids might explain the difference in toxicity of the rosins tested. It is concluded that rosin and zinc are not to be considered inert compounds and that the cytoprotective effects of zinc and its role in dentistry products merit further investigations. PMID- 9335347 TI - Inhibition of the plasma contact activation system of immobilized heparin: relation to surface density of functional antithrombin binding sites. AB - End-point immobilization of heparin to artificial materials gives rise to a surface that prevents triggering of the plasma contact activation system and, presumably as a result thereof, generally has thrombo-resistant properties. The present investigation was undertaken to determine what density of immobilized heparin molecules expressing functionally intact antithrombin binding sites is required to achieve these blood compatible properties. Six different heparin surfaces were prepared on polyethylene tubing and studied in contact with human plasma. The content of bound heparin was the same on all surfaces while the densities of antithrombin binding sites ranged from 1 to 28 pmol/cm2. The surfaces expressing 4 pmol/cm2 or more of specific antithrombin binding sites generated no measurable enzymatic activity in contact with plasma, either on the exposed surfaces or in the plasma phases. Below this level, the degree of activation gradually increased with decreasing densities, and in parallel the thrombo-resistant properties deteriorated. Addition of heparin to the plasma phase reduced the capacity of the heparin surfaces to bind antithrombin, leading to a diminished ability of the surfaces to prevent contact activation. This finding supports the hypothesis that antithrombin is the critical coagulation inhibitor for the suppression of contact activation on end-point immobilized heparin. PMID- 9335346 TI - Ultrastructure of the interface between cultured osteoblasts and surface-modified polymer substrates. AB - Osteoblasts derived from rat bone marrow cells were cultured on surface-modified poly(ethylene terephthalate) films in the presence of ascorbic acid, beta glycerophosphate, and dexamethasone. The surfaces employed for cell culture included the untreated hydrophobic surface and three modified surfaces possessing immobilized phosphate polymer chains, collagen molecules, and a thin hydroxyapatite-deposited layer. They all were produced by photo-induced graft polymerization with subsequent surface modifications of the graft chains. The ultrastructural morphology of the substrate/cell interfaces formed in in vitro osteoblast culture on these substrates was studied by transmission electron microscopy. The osteoblasts cultured for 1 week on the modified surfaces showed rough endoplasmic reticula rich in intracellular space and early matrix production in the extracellular space, irrespective of the surface chemistry. After 2 weeks of culture, osteoblasts exhibited active elaboration of extracellular matrix proteins, mostly composed of collagen, on all the surfaces. A remarkable result observed at this stage was direct deposition of an electron dense, afibrillar layer of 180 nm thickness onto the surface having phosphate polymer chains. This layer became much more electron dense after 3 weeks of culture. Energy dispersive X-ray microanalysis revealed the presence of calcium phosphate in this layer. It was further found that the predeposited hydroxyapatite layer on the phosphate polymer-grafted surface promoted mineral deposition in the extracellular matrix that surrounded cuboid, osteocyte-like cells. PMID- 9335348 TI - Oxidation of ultrahigh molecular weight polyethylene characterized by Fourier Transform Infrared Spectrometry. AB - The effects of processing conditions, sterilization treatment, aging time, and poststerilization aging environment on the oxidation behavior of ultrahigh molecular weight polyethylene (UHMWPE) are examined. Oxidation is monitored by observing changes in the carbonyl peak appearing in Fourier Transform Infrared Spectrometry (FTIR) and is found to be relatively insensitive to processing conditions but strongly influenced by sterilization treatments and aging parameters. Oxygen uptake by UHMWPE increases as a result of gamma or electron beam irradiation and continues to rise during subsequent aging at a rate influenced by the aging environment. A hydrogen peroxide ambient causes more severe oxidation than either air or hyaluronic acid. Control (unsterilized) samples and those sterilized in ethylene oxide are resistant to oxidation under all conditions except hydrogen peroxide aging. PMID- 9335349 TI - Histologic comparison of tibial articular surfaces against rigid materials and artificial articular cartilage. AB - After endoprosthetic replacement of the femoral head, marked pathologic changes of the acetabulum, such as penetration and ulceration, often occur. These changes are caused by the rigid material surface properties of the prosthesis and the lack of damping effects. In this study, we compared the time-dependent changes of tibial articular surfaces with three kinds of femoral implant under loading conditions in dogs. Marked pathologic changes of the menisci and tibial articular cartilage were observed from 8 weeks after implantation with hard material implants, whereas the tibial joint surface against an artificial articular cartilage was still intact 24 weeks postoperatively. These results showed clearly that marked pathologic changes of the articular cartilage against rigid materials occurred and were caused by the surface properties of the counterfaces and high friction coefficients of ceramic and metal materials used. PMID- 9335350 TI - Influence of rapid heating with infrared radiation on RF magnetron-sputtered calcium phosphate coatings. AB - This study evaluated the effect of rapid heating with infrared radiation on the physico-chemical and morphological properties of radio frequent (RF) magnetron sputtered calcium phosphate (Ca-P) coatings. About 2.5 microm thick Ca-P coatings were deposited on titanium disks and cylinders. These specimens were left untreated or were heat treated by infrared radiation at 300, 400, 500, 600, and 700 degrees C for 4, 7, 11, 17, and 24 s. Subsequently, the specimens were immersed in simulated body fluid (SBF) for 1 day, 1 week, and 5 weeks. X-ray diffraction measurements showed that heating at 500 degrees C or higher resulted in an increase of coating crystallinity. In addition, FT-IR measurements revealed the appearance of OH peaks in the spectra of samples treated at 500-700 degrees C. Electron probe microanalysis showed that after 5 weeks of immersion about 40 50% of the coatings heat treated at 500 and 600 degrees C was maintained. The coatings heat treated at 700 degrees C showed no dissolution at all. On the other hand, as-coated and 300 degrees C treated films were dissolved within 1 day. Scanning electron microscopy of the samples showed that directly after heat treatment no apparent cracks were present in the coatings. On the basis of these findings, we conclude that rapid heating with infrared radiation around 600 degrees C is the best heat treatment for RF magnetron-sputtered coatings. PMID- 9335351 TI - Mechanical and biological properties of bioactive bone cement containing silica glass powder. AB - Silica glass powder (SG-P) made by a fusing-quenching method was added as a second filler to a bioactive bone cement consisting of MgO-CaO-SiO2-P2O5-CaF2 apatite and wollastonite containing glass-ceramic powder (AW-P) and bisphenol-a glycidyl methacrylate (Bis-GMA)-based resin, to achieve a higher mechanical strength and better handling properties in use. Five types of cement were used, containing different weight ratios of AW-P/SG-P (Group 1 = 100/0; Group 2 = 75/25; Group 3 = 50/50; Group 4 = 25/75; and Group 5 = 0/100) as filler, to evaluate the effect of SG-P content on the biological, mechanical, and handling properties. The total proportion of filler added to the cements was 85% w/w. The compressive, bending, and tensile strengths and fracture toughness of the cements increased with SG-P content. The viscosity of cements also increased with SG-P content, and every cement could be handled manually. The cements were evaluated in vivo by packing the intramedullary canals of rat tibiae. An affinity index was calculated for each cement; this was the length of bone directly apposed to cement expressed as a percentage of the total length of the cement surface. Histological examination of implanted tibiae for up to 26 weeks showed that the affinity indices decreased with SG-P content and that those of all the cement groups increased with time. At 26 weeks, Groups 1 and 2 had almost identical affnity indices (79% and 75%; no significant difference) but those of the other groups remained at <50%. Group 2 had better mechanical and handling properties than Group 1, and an SG-P content in the filler of no more than 25% w/w did not interfere strongly with the bioactivity of the cement. PMID- 9335352 TI - The role of vitronectin in the attachment and spatial distribution of bone derived cells on materials with patterned surface chemistry. AB - In recent years a central objective of tissue engineering has been understanding the interaction of cells with biomaterial surfaces. In this study we examined the protein adsorption events necessary to control the attachment and the subsequent spatial distribution of bone-derived cells exposed to chemically modified surfaces. Silane chemistry and photolithography techniques were used to create substrates with alternating regions of an aminosilane, N-(2-aminoethyl)-3 aminopropyl-trimethoxysilane (EDS), along side an alkylsilane, dimethyldichlorosilane (DMS), on quartz surfaces. Sera depleted of fibronectin (Fn), vitronectin (Vn), or both were used to determine if these proteins were necessary for the initial attachment and spatial distribution of bone-derived cells exposed to modified surfaces in vitro. The kinetics and mechanisms of the spatial distribution of cells were examined using light microscopy and digital image acquisition and subsequently were analyzed. Compared to complete serum, the use of serum depleted of fibronectin with vitronectin included had minimal effect on the cell attachment, spreading, and spatial distribution on the EDS regions of the surface. However, the use of serum depleted of vitronectin with or without fibronectin included resulted in greatly reduced cell attachment and spreading. Thus the presence of vitronectin was required for the attachment, spreading, and spatial distribution of bone-derived cells exposed to EDS/DMS-patterned surfaces. PMID- 9335353 TI - Crack-growth properties of various elastomers with potential application in small joint prostheses. AB - Silastic small joint spacers for the metacarpophalangeal joint fail catastrophically at a reported rate ranging from 2 to 26%. Although the exact cause of this problem is not known, it is speculated that failure is due to the propagation of flaws generated in the material surface. In addition to wear secondary to bony impingement, these flaws can be introduced through manufacturing, surgical handling, and in vivo frictional wear. In an effort to identify an elastomeric material that will function similarly to Silastic as a self-hinging joint spacer but provide an increased functional life, we have investigated and compared the crack-growth properties of two polyurethanes, ChronoFlex and Medicaflex, and a thermoplastic elastomer, Santoprene, with those of Silastic. The materials were evaluated after sterilization by either ethylene oxide or gamma irradiation in an ASTM standard flexing machine under conditions of high humidity and body temperature both before and after artificially aging. In each case, the materials investigated presented significantly lower crack growth rates than Silastic (p < 0.001). PMID- 9335354 TI - Osteogenic cell cytotoxicity and biomechanical strength of the new ceramic Diopside. AB - Diopside was prepared by sintering a powder compact composed of CaMgSi2O6 at 1573K for 2 h. In order to clarify the biocompatibility of Diopside, the cytotoxicity of Diopside against the osteogenic cell line MC3T3-E1 and the bone Diopside interface strength were examined. On both the 14th and 21st days of incubation of MC3T3-E1 cells with Diopside, ALP activities were not significantly lower than those of the CTRL. TEM photographs of MC3T3-E1 on Diopside after 14 days of incubation showed active secretion of crystals from osteoblast-like cells. Scanning electron microscopic analysis showed that the cells on Diopside formed multiple cell layers similar to those on the CTRL both 14 and 21 days after incubation. These results showed that Diopside had no cytotoxic effect on MC3T3-E1. The pulling test showed that failure loads of Diopside were significantly lower than those of AWGC. Histologically, there was no fibrous tissue or foreign body reaction at the bone interface. SEM-EPMA showed that Diopside had attached to the bone via a calcium-phosphorus layer. SEM back scattered electron imaging showed that the Diopside plate had degraded to a porous state 12 weeks after implantation. These findings indicate that Diopside is a biodegradable ceramic. PMID- 9335355 TI - Activation of blood coagulation at heparin-coated surfaces. AB - It is hypothesized that immobilized heparin exerts a dual role in blood coagulation. On the one hand, the heparinized surface is because of its dense negative charge, thought to initiate the intrinsic pathway of blood coagulation. On the other hand, heparin is known as a potent anticoagulant drug. However, it remains to be seen how much contact-phase activation of factor XI contributes to thrombin formation and how this process is counterbalanced by which of the anti protease activities of immobilized heparin. In the present study we examined the generation of factors XIa, IXa, and Xa, and thrombin in recalcified normal and antithrombin-depleted plasma exposed to polyacrylamide-graft polyurethane (PU) sheets modified by multipoint attachment of two different heparin species. One of them, HAH, contained the specific antithrombin binding sequence and the other one, NAH, had a low affinity for antithrombin and had no anticoagulant activity. Our data demonstrate that in contrast to PU, PU-NAH and PU-HAH are strong mediators of factor XIa and factor IXa formation in normal and antithrombin deficient plasma. Interestingly, compared to PU-HAH and PU-NAH, thrombin formation was only slightly diminished in antithrombin-deficient plasma exposed to PU. In contrast, thrombin formation was dramatically delayed and diminished in normal plasma exposed to PU-HAH. These findings indicate that very low amounts of factor XIa apparently suffice to induce significant amounts of thrombin. In this sense, heparinized surfaces are highly thrombogenic, but our data also indicate that this activity is effectively counterbalanced by the anti-thrombin activity of the immobilized anti-coagulant species of heparin. PMID- 9335356 TI - Compositional dependence of bioactivity of glasses in the system Na2O-K2O-MgO-CaO B2O3-P2O5-SiO2. AB - The bioactivity, i.e., bone-bonding ability, of 26 glasses in the system Na2O-K2O MgO-CaO-B2O3-P2O5-SiO2 was studied in vivo. This investigation of bioactivity was performed to establish the compositional dependence of bioactivity, and enabled a model to be developed that describes the relation between reactions in vivo and glass composition. Reactions in vivo were investigated by inserting glass implants into rabbit tibia for 8 weeks. The glasses and the surrounding tissue were examined using scanning electron microscopy (SEM), light microscopy, and energy-dispersive X-ray analysis (EDXA). For most of the glasses containing < 59 mol % SiO2, SEM and EDXA showed two distinct layers at the glass surface after implantation, one silica-rich and another containing calcium phosphate. The build up of these layers in vivo was taken as a sign of bioactivity. The in vivo experiments showed that glasses in the investigated system are bioactive when they contain 14-30 mol % alkali oxides, 14-30 mol % alkaline earth oxides, and < 59 mol % SiO2. Glasses containing potassium and magnesium bonded to bone in a similar way as bioactive glasses developed so far. PMID- 9335357 TI - Osteoblastic phenotype expression on the surface of hydroxyapatite ceramics. AB - To analyze the bone-bonding property of hydroxyapatite ceramics (HA), composites of rat marrow cells and porous HA were implanted subcutaneously and harvested at 3 to 4 weeks postimplantation. De novo bone formation was observed primarily on the HA surface without fibrous tissue interposition. The HA/tissue interface was analyzed by the observations of thin undecalcified histological sections and fractured surfaces of the implants. The observations were done with a light microscope and a scanning electron microscope (SEM) connected to an energy dispersive spectrometer. The interfacial analyses showed the appearance of osteoblastic cells on the HA surface and that the cells had initiated partially mineralized bone (osteoid) formation directly onto the surface. The osteoid matured into fully mineralized bone, resulting in firm bone bonding to the HA surface. Characterization of osteoblastic cells on the surface was done by determining levels of protein and gene expression of bone Gla protein (BGP, a.k.a. Osteocalcin), i.e., immunohistochemistry and in situ hybridization, respectively. The existence of BGP and mRNA in the cytoplasmic area of the cells confirmed that active osteoblast apposition fabricated primary bone on the HA surface. All of these results indicate the importance of the HA surface in supporting osteoblastic differentiation of marrow stromal stem cells, which leads to firm bone bonding. PMID- 9335358 TI - The initial reactions of graphite and gold with blood. AB - The initial reactions of graphite and gold with blood were investigated by short time exposure to capillary blood and detection of surface-adsorbed plasma proteins and cells with an immunofluorescence technique. Antibodies specific to fibrinogen, complement factors C1q and C3c, prothrombin/thrombin, von Willebrand factor, and platelet- and leukocyte-membrane antigens were used. The fluorescence intensity was quantitated by computer-aided image analysis. Fibrinogen was the most abundant plasma protein immobilized on either surface, and dense populations of platelets adhered to the protein layer. Complement factors and prothrombin/thrombin were found on the graphite surface, localized in fibrin clots or related to platelets. Platelets were activated (expression of selectin CD62) on both surfaces but more extensively so on the gold surface. Activation of polymorphonuclear granulocytes (PMNGs), measured as expression of integrin CD11b, was seen on both surfaces but with different kinetics. On the graphite surface, the CD11b expression was only transient whereas on gold it increased with time. Our data indicate that graphite is more thrombogenic than gold but less inflammatory. PMID- 9335360 TI - Tele-VAD. PMID- 9335359 TI - Behavior of fetal rat chondrocytes cultured on a bioactive glass-ceramic. AB - We examined the behavior of fetal rat chondrocytes cultured on a bioactive glass ceramic containing apatite and wollastonite (A.W.G.C.). Biomaterial surface topography and profiles were evaluated by bidimensional profilometry and revealed a rough surface for the glass-ceramic compared to the plastic coverslips used as controls. Chondrocyte attachment was evaluated by measuring the number of attached cells after one day of culture and by morphological observations. Chondrocytes attached in great numbers to the material surface by means of focal contacts containing vinculin and beta1-integrin. Fluorescent labeling of actin and vimentin revealed a poor spreading of chondrocytes on the bioactive glass ceramic compared to the plastic coverslips, where the cells appeared to adhere intimately to the surface and exhibited polygonal arrays of stress fibers. During the following days of culture, chondrocytes proliferated, colonized the surface of the material, and, finally, on day 10, formed nodular structures composed of round cells separated by a dense extracellular matrix. Furthermore, these clusters of round cells were positive for type II collagen and chondroitin sulfate, both hard markers of the chondrocyte phenotype. In addition, protein synthesis, alkaline phosphatase activity, and proteoglycan production were found to increase gradually during the culture period with a pattern similar to that observed on control cultures. These results demonstrate that the bioactive glass ceramic tested in this study appears to be a suitable substrate for in vitro chondrocyte attachment, differentiation, and matrix production. PMID- 9335361 TI - Adsorption of hepatitis C virus particles onto the dialyzer membrane. AB - It was recently found that the blood level of hepatitis C virus (HCV) RNA is significantly reduced after each dialysis procedure in patients on chronic hemodialysis. This study was designed to elucidate the mechanism for this phenomenon. In two patients with high serum levels of HCV RNA, the filtrate through the dialyzer (TF-alpha, Teijin Co., Tokyo, Japan) was analyzed for viral RNA using the polymerase chain reaction. At the end of dialysis, the filter was washed with saline, and during the saline washing, aliquots were taken for quantification of RNA by the branched DNA method. The HCV core antigen was quantified as a measure of viral particles, and hemoglobin was also measured for correcting for blood contamination. After the clearance of the blood, the filter was washed with guanidinium isothiocyanate, and the recovery of RNA was measured. The filtrate did not contain detectable RNA. The saline washing of the filter after dialysis contained a significant amount of RNA. Washing with guanidinium isothiocyanate of the thoroughly saline washed filter also recovered a significant amount of RNA. During saline washing, the recovery of RNA in the washing was much delayed behind that of hemoglobin, suggesting the adsorption of the former onto the filter membrane. There was a discordant recovery of RNA and HCV core antigen in the washing, the recovery of the former being delayed behind that of the latter. These results indicate that viral particles are adsorbed onto the inner surface of the filter membrane during dialysis. Some of these adsorbed viral particles are perhaps destroyed by hydraulic pressure applied to blood for dialysis. PMID- 9335362 TI - LDL hemoperfusion--a new procedure for LDL apheresis: biocompatibility results from a first pilot study in hypercholesterolemic atherosclerosis patients. AB - Current lipid apheresis techniques can remove atherogenic lipoproteins only from plasma. The initial mandatory separation of plasma and blood cells renders the extracorporeal circuit complex. We recently described the first clinical application of a new lipid adsorber that adsorbs low-density lipoprotein (LDL) and lipoprotein (a) (Lp[a]) directly from whole blood. In continuation of our work, this paper describes the clinical biocompatibility of this new LDL hemoperfusion system. In a 2 center phase II clinical trial, 12 hypercholesterolemic patients suffering from overt coronary or peripheral artery disease were treated once with LDL hemoperfusion. The new LDL adsorber (DALI, Fresenius, St. Wendel, Germany) contained 480 ml of polyacrylate coated polyacrylamide gel. The anticoagulation protocol consisted of an initial heparin bolus followed by an acid citrate dextrose-A (ACD-A) infusion during the treatment. One patient blood volume was treated per session. All sessions were clinically uneventful. No signs of hemolysis or extracorporeal clot formation could be detected, and cell counts remained virtually constant. In a subgroup of patients (n = 4-6), further biocompatibility parameters were studied. Activation of leukocytes (elastase release), thrombocytes (beta-thromboglobulin [beta-TG] extrusion), and monocytes (interleukin (IL)-1beta and IL-6) were minimal. Complement activation (C3a and C5a generation) was negligible. The chosen anticoagulation protocol was both safe (constant ionized calcium levels) and effective (low thrombin-antithrombin formation). In summary, within the scope of a first pilot study, this new LDL hemoperfusion procedure combined the features of excellent clinical tolerance, good biocompatibility, and ease of handling. Phase III clinical trials will have to show whether these encouraging preliminary results can be corroborated in a larger patient population. PMID- 9335363 TI - Effect of polyethylene glycol conjugated bovine hemoglobin in both top-load and exchange transfusion rat models. AB - The purpose of this study was to determine the effect of the hemoglobin based oxygen carrier, polyethylene glycol conjugated bovine hemoglobin (PEG-Hb) on the physiology of the rat. This study was divided into the following 3 parts: pharmacokinetics, cardiovascular, and histopathology. Pharmacokinetic studies evaluated the PEG-Hb circulatory life and the resultant effect on urine composition. Telemetric intravascular blood pressure probes monitored the heart rate and mean arterial pressure. Renal arterial blood flow was determined by intraoperative perivascular ultrasound. Tissue histology was evaluated for both time and model dependent responses. The mean circulatory half-life of PEG-Hb was 17.7+/-0.3 h. Proteinuria and hemoglobinuria were greatly reduced with PEG conjugation. PEG-Hb treated rats produced 8.5 times and 49 times less proteinuria and hemoglobinuria, respectively, than unmodified bovine Hb treated animals. The mean arterial pressure (MAP) in PEG-Hb treated rats was insignificantly different from sham controls undergoing a 30% exchange transfusion while dextran caused an initial reduction and bovine Hb produced a prolonged elevation in the MAP. In these same anesthetized rats, PEG-Hb slightly decreased the heart rate while dextran caused an increase and bovine Hb had no effect. In addition, PEG-Hb was able to maintain the renal arterial blood flow while both Ringer's lactate and bovine Hb caused a reduction in the blood flow. Finally, PEG-Hb treated rats showed a dose and time dependent formation of vacuoles within the renal proximal convoluted tubules and splenic macrophages in both top-load and exchange transfusion models, but no other morphological changes. In conclusion, PEG-Hb had a relatively long vascular persistence that did not cause any significant alterations in the urinalysis, cardiovascular function, or tissue histopathology in the rat. PMID- 9335364 TI - Mechanical stability of the cementless acetabular component with three spikes. AB - We studied the stability of our cementless acetabular component (socket) with 3 spikes in 65 joints of 65 patients who were followed for over 5 years. In 1 case, there was osteolysis around the femoral component (stem) but not around the socket. The movement of the socket and the radiolucent line was observed in 11 cases of osteoarthritis with acetabular hypoplasia (a mean of 43 degrees of the sharp angle) and 3 cases with rapidly destructive coxarthrosis. We could obtain favorable stability in 50 patients, including 38 with osteoarthritis (a mean of 41 degrees of the sharp angle), 8 with aseptic necrosis, 2 with rapidly destructive coxarthrosis, and 2 with rheumatoid arthritis. Our socket is very effective in preventing osteolysis and is expected to provide more stable mechanical stability by arranging an insertion angle (35 degrees of the optimal open angle and 10 degrees of the anteversion angle) and a full bone graft in osteoarthritic patients. PMID- 9335365 TI - Effects of ultrathin silicone coating of porous membrane on gas transfer and hemolytic performance. AB - To assess the effect of an ultrathin (0.2 microm) silicone-coated microporous membrane oxygenator on gas transfer and hemolytic performance, a silicone-coated capillary membrane oxygenator (Mera HP Excelung-prime, HPO-20H-C, Senko Medical Instrument Mfg. Co., Ltd., Tokyo, Japan) was compared with a noncoated polypropylene microporous membrane oxygenator of the same model and manufacturer using an in vitro test circuit. The 2 oxygenators showed little difference in the oxygen (O2) transfer rate over a wide range of blood flow rates (1 L/min to 8 L/min). The carbon dioxide (CO2) transfer rate was almost the same in both devices at low blood flow rates, but the silicone-coated oxygenator showed a decrease of more than 20% in the CO2 transfer rate at higher blood flow rates. This loss in performance could be partly attenuated by increasing the gas/blood flow ratio from 0.5 or 1.0 to 2.0. In the hemolysis study, the silicone-coated membrane oxygenator showed a smaller increase in plasma free hemoglobin than the noncoated oxygenator. The pressure drop across both oxygenators was the same. These results suggest that the ultrathin silicone-coated porous membrane oxygenator may be a useful tool for long-term extracorporeal lung support while maintaining a sufficient gas transfer rate and causing less blood component damage. PMID- 9335366 TI - Blood cardioplegia infusion using a recirculation type circuit generates bradykinin in significant amounts during open heart surgery. AB - The single pass type (SP) of blood cardioplegia is commonly used in North America during open heart surgery. However the recirculation type (RC) of blood cardioplegia is still widely used in other areas including Japan. Infusion blood cardioplegia using the latter technique often decreases the perfusion pressure. To determine the cause for this, blood levels of bradykinin (BK) were measured in cardiopulmonary bypass (CPB) and the 2 types of blood cardioplegic circuits. As the BK levels in the RC cardioplegia (>3,000 pg/ml) rose, the perfusion pressure decreased abruptly with the increase of the BK levels in the CPB circuit. With SP cardioplegia, the BK level was not increased either during cardioplegia (p < 0.009) or CPB (p < 0.009), and the perfusion pressure was not decreased (p < 0.02). We concluded that the SP circuit is superior to the RC one because of the lesser production of BK and thus lesser fluctuation of perfusion pressure. PMID- 9335367 TI - Phenotypic alterations in circulating monocytes induced by open heart surgery using heparinized and nonheparinized cardiopulmonary bypass systems. AB - In this study of 31 patients with coronary bypass surgery, we used flow cytometry to compare heparin-coated and noncoated cardiopulmonary bypass systems on leukocyte activation. We found significant differences between the groups during bypass, with activation of the complement system, measured as elevated levels of C3a desArg, upregulation of granulocyte beta2 integrin (CD11b), and a loss of circulating monocytes when noncoated systems were used. In both groups an early increase in the monocyte cell surface CD62L expression was obvious while the percentage of human leukocyte antigen (HLA)-DR positive monocytes did not alter. The morning after the operation, leukocytosis was present, together with a highly significant reduction in the monocyte expression of CD11b and HLA-DR, indicating the recruitment to the peripheral blood of cells with altered phenotypes. This alteration in phenotype on potent inflammatory cells may be one part of the impaired function of the immunological system reported after major surgery. PMID- 9335368 TI - A biomechanical double sac (pericardium-Pebax) for specially shaped artificial ventricles: a computerized study to evaluate its mechanical and volumetric properties. AB - For original ovoid shaped artificial ventricles, a biomechanical double sac consisting of a biological sac (porcine pericardium) as the blood contact interface and a synthetic sac (Pebax 3533) as the mechanical support to assume systolic-diastolic dynamic constraints was conceived. The volumetric and mechanical properties were assessed with a three-dimensional modeling of Pebax sacs and computerized simulations of their systolic distortions for both right and left ventricular configurations. The stresses and strains of these sacs were represented as quantitative mappings for a maximum end-systolic state and were below the respective threshold values above which the Pebax material is jeopardized for permanent structure impairment. After fatigue tests applied on Pebax strips under the alleged working conditions of Pebax sacs, the material structure was unchanged and maintained its intrinsic mechanical properties. The theoretical maximum stroke volumes were 74.4 cm3 and 62.4 cm3 for the left and right ventricular configurations, respectively. With these mechanical and volumetric features, the biomechanical double sac concept was considered valid and could be provided for a consequent specific total artificial heart. PMID- 9335369 TI - Platelet damage caused by the centrifugal pump: in vitro evaluation by measuring the release of alpha-granule packing proteins. AB - Platelets are more vulnerable to damage than erythrocytes because platelets are easily activated by contact with extracorporeal circuits and by exposure to shear forces. However, the degree of platelet damage caused by centrifugal pumps is unclear. To evaluate platelet damage in different pumping conditions, the rates of increase for specific proteins in platelet alpha-granules, beta thromboglobulin (beta-TG), and platelet factor 4 (PF-4) were measured in both in vitro simulated left ventricular assist device (LVAD) and cardiopulmonary bypass (CPB) conditions and compared with the erythrocyte trauma. A flow of 5.0 L/min with deltaP of 100 mm Hg for LVAD (low pressure head condition) and a flow of 5.0 L/min with deltaP of 350 mm Hg for CPB (high pressure head condition) were investigated. Each condition was tested 4 times for 3 h in a mock circuit with a Capiox (Terumo, Tokyo, Japan) centrifugal pump using fresh human blood. Blood was sampled at 1 h intervals, measuring plasma free hemoglobin (fHb), beta-TG, and PF 4. To evaluate the degree of damage, the rates of increase of fHb, beta-TG, and PF-4 were calculated for each condition as deltafHb/deltaN, deltabeta-TG/deltaN, and deltaPF-4/deltaN where deltafHb is the increase in plasma free hemoglobin, deltabeta-TG is the increase in beta-TG, deltaPF-4 is the increase in PF-4, and deltaN is the increase in the passing number. The passing number is defined in the following equation: N = Qt/V where t is the time, V is the priming volume, and Q is the flow rate. There was no significant difference between the 2 conditions (low pressure head condition versus high pressure head condition) in the rate of increase of fHb (0.0035+/-0.0004 vs. 0.0034+/-0.0010 g/100 L, NS). Contrary to this, the rates of increase for specific proteins in platelet alpha granules in the high pressure head condition demonstrated a significantly higher rate of increase than in the low pressure head condition. The mean rate of increase for beta-TG in the low pressure head condition was 0.22+/-0.03 ng/ml and in the high pressure head condition was 0.51+/-0.05 ng/ml (p < 0.05). The rate of increase for PF-4 in the low pressure head condition was 0.11+/-0.02 ng/ml and in the high pressure head condition was 0.30+/-0.06 ng/ml (p < 0.05). These results suggest that measurements of beta-TG and PF-4 may be more sensitive parameters than hemolysis for evaluating blood cell trauma and that platelets are more vulnerable to mechanical damage by a centrifugal pump than erythrocytes. PMID- 9335370 TI - Unsteady fluid dynamics of several mechanical prosthetic heart valves using a two component laser Doppler anemometer system. AB - Five typical mechanical heart valves (Starr-Edwards, Bjork-Shiley convexo-concave (c-c), Bjork-Shiley monostrut, Bicer-Val, and St. Jude Medical) were tested in the mitral position under the pulsatile flow condition. The test program included measurements of velocity and turbulent stresses at 5 downstream locations. The study was carried out using a sophisticated cardiac simulator in conjunction with a highly sensitive 2 component laser Doppler anemometer (LDA) system. The continuous monitoring of parametric time histories revealed useful details about the complex flow and helped to establish the locations and times of the peak parameter values. Based upon the nondimensional presentation of data, the following general conclusions can be made. First, all the 5 valve designs created elevated turbulent stresses during the accelerating and peak flow phases, presenting the possibility of thromboembolism and perhaps hemolysis. Second, the difference in valve configuration seemed to affect the flow characteristics; third, the bileaflet design of the St. Jude valve appeared to create a lower turbulence stress level. PMID- 9335371 TI - The effect of respiration on the performance of the total artificial heart. AB - An electromechanical driven pusher plate type total artificial heart (TAH) has been developed in the Department of Surgery at Baylor College of Medicine. The control of this TAH is based upon Frank-Starling's law, and the output is likely to be affected by the respiration cycle. The necessity of an averaging system for the pumping to avoid this drifting of the output has been discussed in several papers. In this study, the effect of respiration on the performance of the TAH was evaluated in both in vitro and in vivo conditions. This TAH was driven without an averaging system in its control system. The results indicated that the mean output pressure was stable with minute changes in both the heart rate and outflow. This verifies that the electromechanical driven pusher plate type TAH, which is under development in our laboratory, compensated for the effect of respiration without an averaging system. PMID- 9335372 TI - LDL apheresis in a homozygous familial hypercholesterolemic child aged 4.5. AB - Preliminary experience with the efficacy and safety of dextran sulfate cellulose low-density lipoprotein (LDL) apheresis for the treatment of a 4.5-year-old girl with homozygous familial hypercholesterolemia and coronary artery disease is reported. The decrease of the most atherogenic apolipoprotein B-containing lipoproteins, low-density lipoprotein (LDL) and lipoprotein(a) (Lp [a]), were in the ranges of 63.1-68.7%, and 52.5-58.6%, respectively. The child tolerated LDL apheresis without any clinically significant complications. Therefore, she was submitted to a long-term program of treatment at intervals of 15 days. The experience suggests the possibility of an early beginning of extracorporeal treatment with LDL apheresis in children severely affected by homozygous or double heterozygous familial hypercholesterolemia. PMID- 9335373 TI - Effects of pulsatile flow on gas transfer of membrane oxygenator: MENOX EL-4000 and Gyro C1-E3 pulsatile mode. AB - It is acknowledged that pulsatile flow enhances the gas exchange performance of membrane oxygenators. However, the data for currently developed oxygenators are limited. In this study, the effect of pulsatile flow was assessed utilizing the MENOX EL-4000 oxygenator. The in vitro test was performed following the Association for the Advancement of Medical Instrumentation (AAMI) standards. Pulsatile flow was produced by the Gyro C1-E3 centrifugal pump with periodical changing of the impeller speed. In Study 1, the following 3 groups were created and examined: nonpulsatile flow, pulsatile flow of 40 bpm, and pulsatile flow of 60 bpm. The blood flow rate was maintained at 3 L/min, and the V/Q ratio was 1. In Study 2, four groups were examined, nonpulsatile flow with V/Q = 1, nonpulsatile with V/Q = 2, pulsatile with V/Q = 1, and pulsatile with V/Q = 2. The blood flow rate was maintained at 4 L/min, and the pulse frequency was set at 40 bpm. In study 1, although O2 transfer was not enhanced, CO2 transfer was significantly improved (40-50%) by pulsatile flow, regardless of pulse frequency. Study 2 demonstrated that pulsatile flow resulted in improved CO2 transfer as did higher ventilation (V/Q = 2). Furthermore, even after applying higher ventilation, the pulsatile mode enhanced CO2 transfer more than the nonpulsatile mode. It was considered that the pulsatile mode induced an active secondary flow and enhanced mixing effects, and consequently CO2 transfer was improved. In conclusion, the pulsatile flow significantly enhanced the CO2 transfer of the MENOX oxygenator. It is indicated that applying the pulsatile mode is a unique and effective method to improve the gas exchange performance for a current membrane oxygenator. PMID- 9335374 TI - Anatomical consideration for an implantable centrifugal biventricular assist system. AB - A miniaturized pivot bearing-supported centrifugal blood pump (Gyro PI) has been developed as a long-term biventricular assist system (BiVAS). In this study we determined the anatomical configuration of this system using a bovine model. Under general anesthesia, a left lateral thoracotomy was performed to open the chest. Two Gyro PI-601 pumps for left and right assists were placed in the preperitoneal pocket by a subcostal abdominal incision. The left pump could be placed along the dome of the diaphragm just beneath the apex of the left ventricle. The right pump could be placed next to the left pump. The inlet and outlet ports of both pumps penetrated the diaphragm. The inlet port of the left pump, with a length of 55 mm, was inserted directly into the apex of the left ventricle. A woven Dacron graft (150 mm long, 11 mm inner diameter) was placed between the outlet port of the left pump and the descending aorta. As for the right pump, a 100 mm long and 120 degree angled inflow conduit was placed between the inlet port and the right ventricular infundibulum. The outlet port of the right pump was connected to the main trunk of the pulmonary artery using a 90 mm long, 11 mm inner diameter Dacron graft. We could perform biventricular assistance to confirm the anatomical feasibility of the Gyro implantable centrifugal BiVAS. PMID- 9335375 TI - Canine latissimus dorsi muscle: an apparatus for isometric force measurements. AB - This paper describes a device used to measure the isometric forces generated during electrical stimulation of the canine latissimus dorsi muscle in vivo with a preserved neurovascular supply. This device uses 2 strain gauge force sensors linked to a movable alignment frame to which the muscle is attached. The muscle length is controlled by the application of known weights to the system. The device has a frequency of response of 17.5 Hz and compliance of approximately 0.1 mm N(-1), and its experimental performance was tested in the anesthetized mongrel dog. PMID- 9335376 TI - Aquaporin-2 transfection of Madin-Darby canine kidney cells reconstitutes vasopressin-regulated transcellular osmotic water transport. AB - Water transport across the mammalian collecting tubule is regulated by vasopressin-dependent aquaporin-2 insertion into and retrieval from the apical cell membrane. To establish a cell line that properly expresses aquaporin-2 and its hormone-dependent shuttling, Madin-Darby canine kidney cells were stably transfected with an aquaporin-2 expression construct. Cells of a representative clone (wild-type 10 [WT-10]) were grown on semipermeable supports, and transcellular osmotic water permeability (Pf; in microm/s +/- SEM) was measured. The basal Pf of WT-10 cells, which was lowered with indomethacin, increased from 10.6 +/- 0.8 to 35.7 +/- 1.2 upon incubation with 1-desamino-8-D-arginine vasopressin (dDAVP). This increase coincided with the translocation of aquaporin 2 from an intracellular compartment to the apical membrane. The Pf of untransfected cells (6.5 +/- 0.8) was unchanged by dDAVP. Kinetic studies with WT 10 cells revealed that maximal Pf was obtained within 30 min after dDAVP addition, which remained elevated for at least 90 min. Intracellular cAMP levels peaked within 5 min after dDAVP admission and decreased to basal levels within 45 min. After preincubation with dDAVP, the Pf decreased within 15 min after dDAVP washout and returned to basal levels within 75 min. In conclusion, the WT-10 cells mimic the vasopressin-regulated transcellular water transport and aquaporin 2 translocation as found in collecting duct cells to a great extent, and therefore constitute an in vitro cell model that can be used to study the regulation of transcellular water transport in detail and provide a simplified test system for screening putative aquaporin-2 blockers. PMID- 9335377 TI - Aquaporin-2 in the immature rat: expression, regulation, and trafficking. AB - The immature kidney is resistant to arginine vasopressin. To define the role of aquaporin-2 (AQP-2), the developmental expression of this water channel was studied in rats. AQP-2 levels were lower during early postnatal life, reaching maximal expression at 10 wk of age. Concurrently, urine osmolality increased from 242 +/- 60 to 1267 +/- 311 mosmol/kg. To study the regulation of AQP-2, immature and adult rats were kept on ad libitum intake or were water-deprived. Under normal conditions, AQP-2 levels in the immature rat were significantly lower (52.3 +/- 5.8%, P < 0.001) than in the adult. However, after dehydration the expression increased to adult levels. Interestingly, the increase in AQP-2 observed in the immature kidney was not accompanied by a proportional increase in urine osmolality. To rule out a potential alteration in AQP-2 trafficking, the transport of this water channel was investigated in a group of rats subjected to dehydration, treated with desmopressin acetate (dDAVP), or water loaded. Dehydration and dDAVP stimulated translocation of AQP-2 from intracellular vesicles to the plasma membrane, whereas water loading caused a shift of AQP-2 channels back to intracellular vesicles in both adult and immature animals. In summary, AQP-2 expression and trafficking in the immature kidney is appropriately stimulated by water deprivation and dDAVP. However, urine osmolality remained significantly decreased. From this study, it is concluded that although AQP-2 expression may play a role in the development of urine concentrating abilities, there still is a significant defect, yet to be defined, distal to AQP-2. PMID- 9335378 TI - Expression of the IL-2 receptor on human renal proximal tubular epithelial cells. AB - The high-affinity receptor for interleukin (IL)-2, IL-2R, is composed of three chains, i.e., the alpha, beta, and gamma chains. Previous studies have shown that human proximal tubular epithelial cells (PTEC) express IL-2R alpha during inflammation, such as in the kidneys after renal transplantation or in crescentic glomerulonephritis. Furthermore, in this study it is shown that PTEC in culture produce more complement C3 after stimulation with IL-2, suggesting the presence of a functional IL-2R on PTEC. In these experiments, the binding of IL-2 to PTEC was analyzed. PTEC stimulated with IL-2 exhibited very low binding of digoxigenin labeled IL-2; however, stimulation of PTEC with IL-2, in combination with interferon (IFN)-gamma, resulted in increased binding of the digoxigenin-labeled IL-2. In addition, it was found that IL-2, together with IFN-gamma, enhanced the production of C3 by PTEC from baseline levels of 69.6 +/- 3.4 to 198.6 +/- 3.3 ng of C3 per 10(6) cells. The surface expression of IL-2R alpha and IL-2Rbeta was analyzed using monoclonal antibodies. Although unstimulated PTEC hardly showed detectable expression of these chains, stimulation of PTEC with the combination of IL-2 and IFN-gamma resulted in increases of the index of mean fluorescence for the alpha- and beta-chains of 5.5 +/- 0.5 and 9.7 +/- 0.7, respectively. Reverse transcription-PCR analysis revealed mRNA expression not only for the alpha- and beta-chains, but also for the gamma-chain, after stimulation with the combination of IL-2 and IFN-gamma. This study demonstrates that PTEC express the high affinity IL-2R and that the IL-2R might be involved in the regulation of production of complement component C3 found during renal inflammation. PMID- 9335379 TI - Expression and function of Fas (CD95) on human renal tubular epithelial cells. AB - Renal transplant rejection is characterized by an influx of mononuclear cells in the tubulointerstitial area. Recent studies indicate that tubular damage during graft rejection is dependent, at least in part, on apoptosis. It is thought that apoptosis may be induced by the mononuclear cell infiltrate via the perforin/granzyme or the Fas/Fas ligand pathway. Fas is a 43-kD member of the tumor necrosis factor receptor family, and ligation results in apoptosis of the Fas-bearing cell. The present study analyzes whether Fas is expressed on human tubular epithelial cells in situ and in vitro. It was found that 50 to 70% of the tubules in renal tissue exhibited a positive staining for Fas. To further study the occurrence of Fas on tubular cells, eight different primary proximal tubular epithelial cell (PTEC) lines were analyzed for Fas expression. More than 90% of the PTEC were positive for Fas, and treatment with IFN-gamma resulted in an even higher expression. To determine whether Fas ligation resulted in apoptosis of PTEC in culture, PTEC were incubated with two different anti-Fas antibodies, which were able to induce apoptosis in Jurkat cells. No apoptotic PTEC were observed after Fas ligation, as determined by morphological staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling analysis. Simultaneous CD40 and Fas ligation, or treatment with IFN-gamma before Fas ligation, also did not result in the induction of apoptosis. Fas ligation did not result in proliferation or activation of PTEC, as measured by interleukin-8 production. Apoptotic PTEC could only be detected when the cells were incubated with both anti-Fas antibodies and cycloheximide, resulting in up to 92% apoptotic cells. This study demonstrates that although renal tubular epithelial cells express Fas, they appear to be resistant to Fas-mediated apoptosis, suggesting that Fas-mediated apoptosis does not play a role in the induction of apoptosis during renal transplant rejection. PMID- 9335380 TI - Macrophages promote prosclerotic responses in cultured rat mesangial cells: a mechanism for the initiation of glomerulosclerosis. AB - Glomerulosclerosis is the final outcome of a number of different causes of glomerular injury, during which the structures of the glomerulus are obliterated by extracellular matrix. Accumulating evidence suggests that infiltrating macrophages play a pivotal role in the progression to glomerulosclerosis. The present study defines the role played by macrophages at both cellular and molecular levels in the initiation of the sclerotic process in cultured rat mesangial cells. Macrophage-conditioned medium (MPCM) generated from thioglycollate-elicited, lipopolysaccharide-stimulated macrophages upregulated mesangial cell fibronectin production in a dose- and time-dependent manner, independently of cell proliferation. Immunoprecipitation of metabolically labeled 35S-fibronectin confirmed that the matrix protein was synthesized de novo. The genes for fibronectin and the matrix proteins laminin and collagen IV were also found to be upregulated 2.86 +/- 0.24-, 4.94 +/- 0.17-, and 3.03 +/- 0.31-fold over controls, respectively (P < 0.001). Macrophage modulation of matrix turnover was suggested by an upregulation of both transin and tissue inhibitor of metalloproteinase-1 gene transcription. Transforming growth factor (TGF) beta1, platelet-derived growth factor, tumor necrosis factor (TNF) alpha, or interleukin (IL)-1beta could not be detected in the MPCM per se; however, TGFbeta1 and platelet-derived growth factor AB were found to be secreted into mesangial cell culture supernatants. Secretion was augmented 1.69 +/- 0.16- and 2.28 +/- 0.28 fold, respectively (both P < 0.001), in response to MPCM. Northern blot analysis demonstrated that protein secretion had been preceded by upregulation of the genes for these cytokines (2.2 +/- 0.4-fold [P < 0.001] and 5.7 +/- 1.2-fold [P < 0.004], respectively). Incubation of MPCM with either neutralizing antibody or the growth factor receptor antagonist suramin demonstrated that TGFbeta1 played a significant, although minor, role in MPCM-stimulated fibronectin production. In conclusion, this study provides compelling evidence for a direct role of macrophages in the progression to glomerulosclerosis. PMID- 9335381 TI - Induction of monocyte chemoattractant protein-1 in proximal tubule cells by urinary protein. AB - Cytokines play a pivotal role in synthesis and deposition of extracellular matrix in chronic renal failure (CRF). The proinflammatory properties of monocyte chemoattractant protein (MCP)-1 make it an ideal candidate cytokine for the production of interstitial inflammation in CRF. To investigate the possible role of proteinuria in inducing proximal tubular (PT) MCP-1, MCP-1 mRNA levels were measured by Northern blot and reverse transcription PCR in confluent monolayers of PT cells in primary culture in media containing a variety of proteins. PT cells produced MCP-1 mRNA in response to bovine serum albumin (BSA), delipidated BSA (dBSA; 0.5 to 30 mg/ml), holotransferrin, and apotransferrin (1 to 8 mg/ml). Unstimulated PT cells expressed very low levels of MCP-1 mRNA, detectable by reverse transcription PCR but not by Northern blot. The expression of MCP-1 mRNA reached a peak (sixfold greater than control) within 4 h of exposure to dBSA and was maintained for at least 24 h with continued exposure. Removal of dBSA from the media led to a rapid decline in MCP-1 mRNA expression. dBSA-induced MCP-1 expression was inhibited by lysine, an inhibitor of protein uptake, and reproduced by dBSA purified by gel and size-selective filtration. dBSA influenced MCP-1 expression at the level of transcription and probably translation, as evidenced by abrogation of MCP-1 by actinomycin D and superinduction with the protein synthesis inhibitor cycloheximide. The concentration of MCP-1 protein in response to dBSA added to the apical surface of PT cells was 2.4-fold greater in basolateral than in apical media, indicating basolateral secretion of MCP-1 protein. In summary, PT cell MCP-1 mRNA and protein expression are upregulated by albumin and transferrin, in concentrations similar to those of proteinuric urine. This effect could explain the link between proteinuria and interstitial inflammation in CRF. PMID- 9335382 TI - Association between vitamin D receptor gene polymorphism and relative hypoparathyroidism in patients with chronic renal failure. AB - To study the influence of vitamin D receptor (VDR) gene polymorphism on parathyroid cell function in chronic renal failure, 85 patients who had serum PTH levels <12 pmol/L (the low intact PTH [iPTH] group) and 46 patients who had serum iPTH levels >60 pmol/L (the high iPTH group) were selected out of a total dialysis population of 170 individuals. As a result of subsequent exclusions based on several criteria in both groups (diabetic patients, serum aluminum levels, serum calcium levels, and time on dialysis), the final low iPTH group consisted of 34 patients and the final high iPTH included 32 patients. A healthy control population (n = 120) and 162 of the 170-patient dialysis population served as control groups. VDR gene polymorphism was determined by digestion with the BsmI enzyme and single-strand conformation polymorphism analysis of PCR amplified fragments. Serum iPTH levels were lower in patients with the BB genotype than in those with the Bb or bb genotype, both in the total dialysis population and when the various exclusion criteria were applied. No differences in genotypic and allelic frequencies were found between the healthy control population and the high iPTH group. However, the genotypic distribution was significantly different in the low iPTH group of patients before and after applying all exclusion criteria (P = 0.037 and P = 0.018, respectively). In the final selected population, the bb genotype was less frequent in the low iPTH group than in the total dialysis population (14.7% versus 36.4%; odds ratio, 0.3; confidence interval, 0.11 to 0.82; P = 0.01). Conversely, the BB genotype was over-represented in the low iPTH group (23.3% versus 19.7%; odds ratio, 1.9; confidence interval, 0.85 to 4.3; P = 0.1). In addition, the bb genotype and the b allele frequencies were lower in the low iPTH group than in the high iPTH group (14.7% versus 34.4%, P = 0.06, and 41.2% versus 60.9%, P = 0.02, respectively), and the BB genotype and the B allele were significantly more frequent in the low PTH group than in the high iPTH group (32.3% versus 12.5%, P = 0.05, and 58.8% versus 39.1%, P = 0.02, respectively). Thus, VDR gene polymorphism influences parathyroid function in chronic renal failure. PMID- 9335383 TI - Bone density and skeletal metabolism in patients with orthotopic ileal neobladder. AB - The interposition of a bowel segment as a bladder substitute into the urinary tract may result in impaired calcium metabolism. We studied 25 male patients (aged 45 to 77 yr) who had undergone a Vescica Ileale Padovana (VIP) reconstruction following cystectomy 29 to 75 mo before. Bone mineral density of the spine and femur was measured by dual x-ray absorptiometry. Blood and 24-h urine samples were analyzed for the main parameters of bone metabolism. Sixteen healthy men were enrolled as a control group. Although blood pH did not differ between patients and control subjects, VIP subjects showed lower levels of plasma HCO3- (P < 0.005) and higher serum chloride (P < 0.001). Bone alkaline phosphatase was higher (P < 0.001), and urine calcium, phosphate, and creatinine levels were lower in VIP patients (P < 0.01, P < 0.01, and P < 0.05, respectively). Bone mineral density at the femoral neck (P < 0.03) and Ward's triangle (P < 0.05) was decreased in VIP patients. When subdivided according to time since operation, patients who had the ileal neobladder implanted for a shorter period of time showed lower blood pH (P < 0.03) and urine calcium (P < 0.05) levels and higher urinary hydroxyproline (P < 0.02). Duration of the ileal neobladder was positively correlated with PTH (r = 0.46, P < 0.03) and blood pH (r = 0.47, P < 0.02). Furthermore, pH values were positively correlated with urine calcium (r = 0.48, P < 0.02). In conclusion, in patients with ileal neobladder, a mild metabolic acidosis is responsible for an increased bone turnover and lower bone mass. Moreover, a decrease over time in the absorption capacity of the ileal pouch might result in calcium malabsorption, which represents an additional risk factor for reduced bone mass in these patients. PMID- 9335384 TI - Identification of patients with autosomal dominant polycystic kidney disease at highest risk for end-stage renal disease. AB - To identify those potential factors that, early in the course of disease, mark a population of patients with autosomal dominant polycystic kidney disease (ADPKD) who have worse renal survival, survival analysis and risk ratio calculation for 1215 ADPKD patients were performed. Survival times were calculated as time to dialysis, transplantation, or death. Risk ratios were calculated using the Cox proportional hazards model. Three hundred eighty-eight patients entered end-stage renal disease and 205 patients died. ADPKD2 subjects had longer renal survival than ADPKD1 subjects (median survival, 68 versus 53 yr; P < 0.0005; risk ratio, 2.5). Women had significantly better renal survival than men (56 versus 52 yr; P < 0.0001; risk ratio, 1.6). Subjects who were diagnosed before age 30 and those who developed hypertension before age 35 had worse renal survival than those subjects who were diagnosed after age 30 or those who remained normotensive after age 35, respectively (age of diagnosis: 49 versus 59 yr; P < 0.0001; risk ratio, 3.2; hypertension: 51 versus 65 yr; P < 0.0001; risk ratio, 4.4). Similarly, those who had an episode of gross hematuria before age 30 had a worse renal outcome than those who did not (49 versus 59 yr; P < 0.0001; risk ratio, 2.6). We have also calculated risk ratios for a combined model. When therapeutic interventions become available for this disease, these populations with high risk ratios should be considered for such interventions. PMID- 9335385 TI - Family history and risk of kidney stones. AB - Kidney stones develop more frequently in individuals with a family history of kidney stones than in those without a family history; however, little information is available regarding whether the increased risk is attributable to genetic factors, environmental exposures, or some combination. In this report, the relation between family history and risk of kidney stone formation was studied in a cohort of 37,999 male participants in the Health Professionals Follow-up Study. Information on family history, kidney stone formation, and other exposures of interest, including dietary intake, was obtained by mailed questionnaires. A family history of kidney stones was much more common in men with a personal history of stones at baseline in 1986 than in those without a history of stones (age-adjusted prevalence odds ratio, 3.16; 95% confidence interval [CI], 2.90 to 3.45). During 8 yr of follow-up, 795 incident cases of stones were documented. After adjusting for a variety of risk factors, the relative risk of incident stone formation in men with a positive family history, compared with those without, was 2.57 (95% CI, 2.19 to 3.02). Family history did not modify the inverse association between dietary calcium intake and the risk of stone formation. These results suggest that a family history of kidney stones substantially increases the risk of stone formation. In addition, these data suggest that dietary calcium restriction may increase the risk of stone formation, even among individuals with a family history of kidney stones. PMID- 9335386 TI - The influence of volume depletion and central hypovolemia on the plasma concentration of parathyroid hormone in dialysis patients. AB - Because changes in extracellular volume during dialysis cause reflex neurohonnonal changes that may influence parathyroid hormone (PTH) release independently of calcium, the influence of isotonic volume depletion (by isolated ultrafiltration) and central hypovolemia (70 degrees tilt) on serum PTH1-84 was studied in 16 hemodialysis patients. Tilting was performed in volume depleted state, i.e., immediately after hemodialysis. In the control study, patients underwent sham ultrafiltration (UF = 0) and after dialysis maintained the supine position for the same length of time they remained in the tilt position in the active experiment. Isolated ultrafiltration (-2.3 +/- SEM 0.3 L) caused a 21% fall in mean arterial pressure (from 101 +/- 6 to 80 +/- 6 mmHg, P < 0.01), a fall that was accompanied by a marked increase in plasma catecholamine levels (norepinephrine P < 0.001, epinephrine P < 0.025), in plasma renin activity (P < 0.001) and in plasma arginine vasopressin (P < O.001). Atrial natriuretic factor showed a slight reduction, whereas the plasma endothelin-1 level did not change. Serum Ca showed the expected, hemoconcentration-dependent rise (from 4.1 +/- 0.1 to 4.4 +/- 0.1 meq/L, P < 0.01). Interestingly, UF caused a marked rise in plasma PTH1-84 concentration (from 252 +/- 62 to 335 +/- 72 pg/ml, P < 0.01). UF-induced changes in serum PTH1-84 were related to norepinephrine changes (r = 0.57) as well as to plasma renin activity (r = 0.50). After hemodialysis, tilting induced a pronounced rise in serum PTH1-84 (from 102 +/- 29 to 200 +/- 55 pg/ml), and these changes were slightly related to plasma epinephrine (r = 0.49) but independent of other parameters. In the control experiment, neither sham UF nor recumbency modified serum PTH. In hemodialysis patients, serum PTH is sensitive to changes in extracellular and central blood volume of magnitude sufficient to decrease arterial pressure. Avoiding marked volume stimuli might help to refine the interpretation of the Ca/PTH curves during hemodialysis in these patients. PMID- 9335387 TI - Hypercalcemia during pulse vitamin D3 therapy in CAPD patients treated with low calcium dialysate: the role of the decreasing serum parathyroid hormone level. AB - Oral pulse therapy with vitamin D is effective in suppressing parathyroid hormone (PTH) secretion in continuous ambulatory peritoneal dialysis patients with secondary hyperparathyroidism (2'hpt). However, this treatment often leads to hypercalcemia. The goals of the study were: (1) to examine whether the incidence of hypercalcemia decreases when dialysate calcium is reduced from 1.25 to 1.0 mmol/L; (2) to determine the relative role of the factors involved in the pathogenesis of hypercalcemia; and (3) to study the efficacy of a low oral pulse dose of alfacalcidol in preventing the recurrence of 2'hpt. Fourteen continuous ambulatory peritoneal dialysis patients with 2'hpt were treated with pulse oral alfacalcidol and calcium carbonate and dialyzed with a 1.0-mmol (n = 7) or a 1.25 mmol (n = 7) dialysate calcium. The response rate (87%) and the incidence (71%) and severity of hypercalcemia were similar in both groups. In the early response stage, PTH decreased by 70% in both groups, and serum ionized calcium (iCa) increased from 1.18 +/- 0.02 to 1.27 +/- 0.04 mmol/L (P < 0.005) in the 1.0 group and from 1.19 +/- 0.02 to 1.29 +/- 0.02 mmol/L in the 1.25 group (P < 0.005). Nine of the 12 responders had a further decrease in serum PTH, which was associated with an additional increase in iCa from 1.28 +/- 0.02 to 1.47 +/- 0.04 (P < 0.005). Multivariate analysis showed that the early increase in iCa was positively correlated with alfacalcidol dosage (r = 0.69). In contrast, the late increase in iCa was mostly accounted for by the decrease in serum PTH (r = 0.93). This occurred while calcium carbonate, alfacalcidol dosage, and serum 1,25 hydroxy D3 remained unchanged compared with the early response stage. Finally, an alfacalcidol dose of 1 microg twice weekly was unable to maintain serum PTH at an adequate level in the long term. These data show that a reduction in dialysate calcium from 1.25 to 1.0 mmol does not reduce the occurrence of hypercalcemia and suggest that lowering serum PTH reduces the ability of the bone to handle a calcium load within a few weeks, thus causing hypercalcemia. PMID- 9335388 TI - Lack of removal of calcitriol during hemodialysis procedures. AB - In this study, an in vitro study with blood was performed to determine whether calcitriol (Calcijex) is either bound or removed from blood by dialyzer circuits. The study utilized five different dialyzers, each with a widely varying membrane type, and standard dialysate blood lines. One liter of fresh whole bovine blood was injected with 0.6 microg calcitriol and was dialyzed against 5 L of acetate based dialysate for 180 min. By measurement of changes in blood and dialysate levels of calcitriol, there was no evidence of any calcitriol dialytic clearance and no evidence of calcitriol binding to the blood side components for any of the dialysis circuits. The results of this study indicate that calcitriol can be injected intravenously at any time during a dialysis procedure, without significant removal of the drug from the blood by dialyzer clearance or by binding to materials in the dialyzer circuit. PMID- 9335389 TI - The cost effectiveness of mycophenolate mofetil in the first year after primary cadaveric transplant. U.S. Renal Transplant Mycophenolate Mofetil Study Group. AB - Mycophenolate mofetil (MMF) has been shown to reduce the incidence of acute graft rejection in three controlled trials of cadaveric renal transplantation. In a U.S. trial using quadruple sequential induction therapy as control, the MMF 2-g treatment group had an acute rejection rate 40.6% lower than control in the first posttransplant year (27.9% MMF-treated versus 47.0% control). The purpose of this analysis is to evaluate the economic implications of these clinical differences. The analysis relies on resource use data from the trial and other sources. Medical costs were estimated using a societal perspective and excluded the cost of the transplant procedure and organ acquisition. The two groups were compared in terms of treatment for acute rejection and opportunistic infection, graft survival, dialysis use, and maintenance immunosuppression. The results suggest that, on average, when compared with standard therapy, patients treated with MMF are likely to have lower rejection-related treatment costs because of a lower incidence of rejection ($6237 versus $3702), lower dialysis and graft failure costs because of improved graft survival ($20,104 versus $16,972), no difference in opportunistic infection treatment costs ($1962 versus $1962), and higher additional immunosuppression costs ($855 versus $5170). Taken together, these results suggest that patients treated with MMF are, on average, likely to have slightly lower first-year costs ($29,158 versus $27,807) compared with control, indicating that MMF treatment is cost-effective in the first year. These results remained stable under sensitivity analyses, with plausible variation in the rates of acute rejection, graft survival, and infection. PMID- 9335390 TI - The hyponatremic patient: practical focus on therapy. PMID- 9335391 TI - Adenylate cyclase activity along the rabbit nephron as measured in single isolated segments. 1975. PMID- 9335393 TI - The Kidney Council of the American Heart Association needs support to maintain a good thing. PMID- 9335392 TI - Managed care, capitation, and the future of nephrology. AB - Within the next decade, it is predicted that more than 90% of the United States population will receive its health insurance through managed care. Capitation will be the reimbursement mechanism to health care providers as the major way of controlling costs. Currently, managed care has had little experience with capitation payments for chronically ill patients, who consume large financial and physical resources. The end-stage renal disease (ESRD) population represents a vulnerable group of patients, and their care may be compromised in a capitated environment. Nephrologists will need to serve as advocates for ESRD patients through a mechanism of quality of care, driven by a continuous quality improvement model. Cost-effective delivery of care will occur as nephrologists join together to form Independent Practice Associations (IPAs). In this article, the role of a nephrologist in a capitated environment is outlined in detail, and background for the basis of managed care growth is provided as a framework for understanding the change in our health care delivery system. After formation of a nephrology IPA, there will most likely be a linkage with a management service organization (MSO). A business plan driven by the highest principles will allow nephrologists to work together as a cohesive force in accepting global risk capitated contracts. The starting point is for ESRD care, and the future includes pre-ESRD care. PMID- 9335394 TI - Severe hyperparathyroidism associated with prolonged hungry bone syndrome in a renal transplant recipient. AB - Although widely believed to resolve within 6 to 12 months of successful renal transplantation, hyperparathyroidism may persist or develop after renal transplantation and eventually require parathyroidectomy. Avid calcium retention by demineralized bones (hungry bone syndrome) is well-recognized after parathyroidectomy and usually resolves after a few weeks. This report documents the case of a renal transplant recipient with persistent hyperparathyroidism who developed a pathological fracture of the pelvis and required parathyroidectomy 1 year after transplant and then manifested severe and prolonged hungry bone syndrome lasting for more than 20 months postoperatively. The clinical features and treatment of hyperparathyroidism in renal transplant recipients are discussed, as are diagnosis, pathogenesis, and management of hungry bone syndrome. Recognition of renal transplant recipients at greater risk for severe hungry bone syndrome should permit earlier and more aggressive management of this sometimes protracted complication of parathyroid surgery. PMID- 9335395 TI - Diffuse proliferative glomerulonephritis and acute renal failure associated with acute staphylococcal osteomyelitis. AB - A 72-year-old man developed acute renal failure after coronary bypass surgery that had been complicated by sternal osteomyelitis caused by the Staphylococcus aureus bacterium. Bacteremia and sepsis were not present. Renal biopsy demonstrated a florid, diffuse, proliferative glomerulonephritis with glomerular immune complex deposition. Management included hemodialysis, prolonged antibiotic therapy, and repeated surgical debridement. Spontaneous recovery of renal function occurred after eradication of infection and final surgical wound repair. The relationship between acute bacterial infections and glomerulonephritis and, in particular, the causal role of staphylococcal antigens in the pathogenesis of such lesions is discussed. PMID- 9335396 TI - Cisplatin-induced renal toxicity: possible reversal by N-acetylcysteine treatment. AB - Cisplatin administration is frequently associated with renal insufficiency and tubular dysfunction. In this article, a case of massive cisplatin overdose as a result of an accidental substitution for carboplatin is reported. The patient developed oliguric renal failure, hepatotoxicity, ototoxicity, peripheral neuropathy, blindness, and severe myelosuppression. A therapeutic trial with N acetylcysteine to reverse renal toxicity was attempted. This article reviews the literature on cisplatin nephrotoxicity, with an emphasis on possible mechanisms involved, and suggests therapy for its modification. PMID- 9335397 TI - Basal muscarinic activity does not impede beta-adrenergic activation in rabbit hearts in controls or thyroxine-induced cardiac hypertrophy. AB - We tested the hypothesis that basal myocardial muscarinic receptor activity acts as a "brake" on beta-adrenergic activation and that this effect would be greater in hearts subjected to thyroxine (T4)-induced (0.5 mg/kg for 16 days) hypertrophy due to an increase in muscarinic receptor density. Twenty control and 20 T4 treated open-chest anesthetized New Zealand white rabbits were given isoproterenol (0.5 microg/kg/min, 10 min i.v.) and/or atropine (3 mg/kg bolus). Coronary blood flow (radioactive microspheres), aortic and left ventricular (LV) pressure, and wall thickening of the LV free wall were recorded. Hearts were quickly excised and stored in liquid nitrogen. Cyclic guanosine monophosphate (GMP) and cyclic adenosine monophosphate (AMP) were determined by radioimmunoassay. T4 increased heart weight/body weight ratio, blood pressures, and the first derivative of the maximal rate of increase of LV systolic pressure (dP/dt[max]). Isoproterenol increased heart rate in both groups. Atropine had no effects on hemodynamic parameters either alone or after stimulation with isoproterenol. At this dose, atropine completely blocked the depressant effects of acetylcholine (10 microg/kg). Isoproterenol increased the maximal time derivative of wall thickening (dWT/dt[max]) in control (from 11.0 +/- 1.0 to 16.4 +/- 1.5 mm/s) but not in T4 animals. T4 increased subepicardial (EPI) and subendocardial (ENDO) coronary blood flow. Isoproterenol increased coronary flow (control: EPI, from 173 +/- 11 to 346 +/- 28 ml/min/100 g; ENDO, from 197 +/- 15 to 364 +/- 30 ml/min/100 g; T4: EPI, from 314 +/- 45 to 459 +/- 43 ml/min/100 g; ENDO, from 339 +/- 48 to 458 +/- 43 ml/min/100 g). Cyclic AMP levels were higher in T4 animals. Isoproterenol increased cyclic AMP (control: EPI, from 540 +/- 82 pmol/g to 1,096 +/- 110; ENDO, 596 +/- 58 to 1,050 +/- 145 pmol/g; T4: EPI, from 882 +/- 107 pmol/g to 1,319 +/- 222; ENDO, from 954 +/- 134 to 1 ,409 +/- 261 pmol/g). Atropine, alone or after stimulation with isoproterenol, had no effect on coronary flow or cyclic AMP in either group. Cyclic GMP levels were unaffected by T4-induced hypertrophy or by any of the treatments in either group. Thus it appears that basal muscarinic activity does not significantly influence function or signal transduction either at baseline or during beta-adrenergic stimulation in controls or in T4-induced hypertrophy. PMID- 9335398 TI - Effects of long-term amiodarone treatment on ventricular-fibrillation vulnerability and defibrillation efficacy in response to monophasic and biphasic shocks. AB - Antiarrhythmic drugs, most notably amiodarone, are often used to combat life threatening tachyarrhythmias simultaneous with implantable cardioverter defibrillators. However, the effects of long-term amiodarone treatment on ventricular fibrillation (VF) vulnerability and the defibrillation threshold (DFT) remain incompletely understood. VF vulnerability and the DFF for monophasic and biphasic shocks were studied in 10 isolated perfused hearts of rabbits treated over the long term with amiodarone (50 mg/kg/day orally for 28 days) before the experiment. The results were compared with those of a control group (n = 10). Monophasic action potentials were recorded from 10 sites simultaneously to determine ventricular activation and repolarization. Myocardial tissue concentrations were 17.1 +/- 14.8 microg/g for amiodarone and 4.6 +/- 4.4 microg/g for desethylamiodarone. Amiodarone treatment prolonged action-potential duration by 12.9 ms (p = 0.025) and ventricular repolarization by 16.5 ms (p = 0.03) without changing ventricular activation and dispersion of repolarization. Amiodarone treatment caused a rightward shift of the vulnerable window for monophasic and biphasic shocks by 13-17 ms (p < 0.05). The width of the vulnerable window, the upper (ULV) and lower (LLV) limits of VF vulnerability, and the DFT remained unchanged. The fact that ULV and DFT remained unchanged suggests that the ULV still may be valid surrogate for the DFT during long-term amiodarone therapy. PMID- 9335399 TI - Effects of nitroprusside and nicorandil on catecholamine secretion and calcium mobilization in cultured bovine adrenal chromaffin cells. AB - The effects of nitroprusside and nicorandil on catecholamine secretion and free intracellular Ca2+ ([Ca2+]i) mobilization in bovine adrenal chromaffin cells were studied to evaluate the role of the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway in the regulation of a [Ca2+]i-dependent secretory process. These effects were compared with those of nifedipine, a typical voltage dependent Ca2+ channel blocker. Carbachol produced a rapid increase followed by a sustained increase of [Ca2+]i (Ca2+ transient) in cultured bovine adrenal chromaffin cells. Both nitroprusside and nicorandil accelerated the decrease in [Ca2+]i without changing the peak values of the initial [Ca2+]i increase of Ca2+ transient. These drugs, however, did not affect carbachol-induced catecholamine secretion, suggesting that secretion is related to the initial [Ca2+]i increase and not to a late sustained [Ca2+]i increases. However, nifedipine reduced the peak and duration of carbachol-induced [Ca2+]i increases and decreased the secretion of catecholamines. Diethylamine/NO complex, an NO donor, and dibutyryl cGMP produced similar changes in Ca2+ transient and did not alter catecholamine secretion, suggesting that the effects of nitroprusside and nicorandil were mediated by the NO/cGMP pathway. These results indicated that the pattern of Ca2+ transient, especially the initial increase in [Ca2+]i, is important in secretion of catecholamine. PMID- 9335400 TI - Diltiazem inhibits the late increase in extracellular potassium by maintaining glycolytic ATP synthesis during myocardial ischemia. AB - During myocardial ischemia, the extracellular potassium concentration increases in a triphasic pattern, an initial early increase, a constant phase, and a late increasing phase. The aim of this study was to determine whether diltiazem inhibits the late increasing phase by maintaining glycolytic adenosine triphosphate (ATP) synthesis in ischemic myocardium. The extracellular potassium concentration and pH were measured simultaneously with ion-sensitive electrodes during 30 min of global ischemia in isolated guinea-pig hearts. In the control hearts, the late increasing phase occurred 13 min after the onset of ischemia when the change in extracellular pH had reached a plateau. There was a sharp increase in the myocardial lactate level in the control hearts, which was maintained for approximately 8 min after the onset of ischemia. Iodoacetate (1 mM) led to a ATP depletion and rapid accumulation in extracellular potassium shortly after the onset of ischemia without a decrease in extracellular pH. The preischemic treatment with diltiazem (3 microM) reduced cardiac function both before ischemia and during the early period of ischemia. Diltiazem almost completely abolished the late increasing phase with a continuous decrease in extracellular pH throughout the ischemic period. The myocardial lactate level in the diltiazem-treated group increased sharply between 2 and 15 min after the onset of ischemia. The myocardial ATP level was preserved throughout the ischemic period. This study shows that diltiazem inhibits the late increasing phase in extracellular potassium by maintaining glycolytic ATP synthesis during ischemia. PMID- 9335401 TI - Diversity in the renal hemodynamic effects of dihydropyridine calcium blockers in spontaneously hypertensive rats. AB - Effects of dihydropyridine (DHP) Ca2+ channel blocker (CaB) on glomerular hemodynamics are controversial. We examined the effects of two DHP derivatives, benidipine hydrochloride (BDP) and nifedipine (NDP), on glomerular hemodynamics by an in vivo micropuncture method by using spontaneously hypertensive rats (SHRs). Systemic bolus infusion of BDP (4 microg/kg) or NDP (250 microg/kg) elicited comparable decreases in mean arterial blood pressure (MAP). The proximal stop-flow pressure (Psf), an indicator of glomerular capillary pressure, revealed significant decreases in BDP but nonsignificant increases in NDP. To minimize the influence of MAP or other systemic events, we monitored Psf during perinephron infusion of CaB and observed significant increases in Psf during 10(-3) M NDP perfusion and nonsignificant changes with 10(-3) M BDP. Moreover, the stability of Psf during alteration of renal perfusion pressure was significantly impaired in the nephron treated with topical NDP. These findings support the notion that CaB has diverse effects on glomerular hemodynamics, and such effects may in part be the result of different pharmacologic actions on the renal microvasculature. PMID- 9335402 TI - Infarct size-reducing effect of ischemic preconditioning is related to alpha1b adrenoceptors but not to alpha1a-adrenoceptors in rabbits. AB - In rabbits and rats, both stimulation of alpha-adrenoceptors and ischemic preconditioning (PC) reduce infarct size. Activation of alpha1b-adrenoceptors play an important role in the PC effect on ventricular function in rats. However, the alpha1-adrenoceptors have not been reported to be related to the PC effect in rabbits, because the infarct size-reducing effect of PC is not blocked by the nonselective alpha-adrenoceptor antagonist, phenoxybenzamine (POB) or by the alpha1-adrenoceptor antagonist, BE2254. However, we speculated that alpha1b adrenoceptors but not alpha1a-adrenoceptors may be related to the infarct size reducing effect of PC in rabbit hearts. Thus we examined in rabbits whether the alpha1b-adrenoceptor blocker chloroethylclonidine (CEC), the alpha1a-adrenoceptor blocker 5-methylurapidil (5-MU), the selective alpha1-adrenoceptor antagonist bunazosin (BN), and the nonselective apha-adrenoceptor antagonist phenoxybenzamine (POB) can block the PC effect on infarct size. Eighty-eight anesthetized open-chest Japanese white male rabbits were subjected to 30-min coronary occlusion and 48-h reperfusion. In five PC groups, the rabbits were subjected to a single 5-min occlusion and 5-min reperfusion before 30-min sustained ischemia. In the PC groups, those with CEC (3 mg/kg, n = 10), 5-MU (3 mg/kg, n = 10), BN (0.3 mg/kg, n = 10), POB (4 mg/kg, n = 10), or placebo saline (n = 10) were pretreated before PC. In the non-PC groups, those with CEC (3 mg/kg, n = 7), 5-MU (3 mg/kg, n = 7), BN (0.3 mg/kg, n = 7), POB (4 mg/kg, n = 7), or placebo saline (n = 10) were pretreated before 30-min sustained ischemia. After a 48-h reperfusion, the infarct size was measured histologically and expressed as a percentage of the area at risk. PC caused a marked reduction of infarct size as compared with the non-PC control (10 +/- 3% vs. 42 +/- 2%; p < 0.05). The PC effect was completely blocked by CEC (36 +/- 2%) and by BN (42 +/- 4%) but not by 5-MU (14 +/- 1%) or POB (13 +/- 2%). None of the drugs by itself affected the infarct size. Stimulation of alpha1b-adrenoceptors but not of alpha1a-adrenoceptors during PC plays an important role in the PC effect on infarct size. This may explain the previous confusion concerning the PC blocking effect of various alpha1-blockers. PMID- 9335403 TI - Effects of bradykinin and icatibant on renal hemodynamics in conscious spontaneously hypertensive and normotensive rats. AB - We investigated the effects of bradykinin (BK) and icatibant (HOE 140), a highly selective bradykinin-B2-receptor antagonist, on mean arterial blood pressure (MAP), heart rate (HR), renal blood flow (RBF), and renal vascular resistance (RVR) in conscious Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). Experiments were performed in conscious male WKY rats and SHRs instrumented over the long term with arterial and venous catheters and a transit time flow probe for measurement of RBF. In WKY rats (n = 16), intraaortic (i.a.) bolus injections of BK (0.1, 1.0, and 10 microg) produced dose-dependent decreases in MAP and RBF with reciprocal increases in RVR. Intrarenal (i.r.) injections of BK (10 microg; n = 6) induced the same hemodynamic response pattern, although the increase in RVR was higher compared with i.a. injections (p < 0.05). Neither vasopressin V1-receptor nor alpha1-adrenoceptor blockade had an effect on the renal vasoconstrictor responses on i.a. BK. The i.a. injections of icatibant (0.1, 1.0, 5.0, and 10 microg; n = 6-10 for each dose) led to a dose dependent blockade of the hemodynamic responses to BK (10 microg, i.a.). Icatibant (10 microg, i.a) had no effect on resting MAP and HR but induced a biphasic response in RBF and RVR with significant changes compared with basal values (p < 0.05). In SHRs (n = 9), after injection of increasing i.a. doses of BK (0.1, 1.0, and 10 microg), dose-dependent decreases in MAP were proportionately greater compared with those in WKY rats. In contrast to WKY rats, RBF and RVR exhibited a biphasic response pattern on BK in SHRs. Neither vasopressin V1-receptor nor alpha1-adrenoceptor blockade had an effect on the renal vasoconstrictor responses on i.a. BK. The i.a. injections of icatibant (10 microg) almost completely blocked the hemodynamic responses on BK in SHRs (n = 13). Icatibant (10 microg, i.a.) itself induced an increase in resting MAP and HR (p < 0.05) and a biphasic response in RBF and RVR with significant changes of basal values (p < 0.05). Our results provide evidence that BK exhibits renal vasoconstrictor and vasodilator properties in vivo, both mediated by B2 receptors. Furthermore, we demonstrated that SHRs display an increased B2 receptor-mediated vasodilatory responsiveness to BK. Finally we showed that blockade of B2 receptors leads to an increase of MAP in SHRs in contrast to WKY rats, suggesting an important role of the kallikrein/kinin system in the regulation of high blood pressure in SHRs. PMID- 9335404 TI - Soluble E-selectin and intercellular adhesion molecule-1 plasma levels increase during acute myocardial infarction. AB - Previous studies have shown that adhesion molecules play a crucial role in leukocyte-endothelium interactions that occur during myocardial ischemia and reperfusion. We assessed the plasma levels of the soluble form of E-selectin (sE selectin) and intercellular adhesion molecule-1 (sICAM-1) in 15 patients with acute myocardial infarction (AMI) and in 15 controls with chronic stable angina. In patients with AMI, the levels of sE-selectin and sICAM-1 increased significantly during the first 8 h after infarction and subsequently decreased. Soluble E-selectin levels were inversely related to the peak plasma levels of creatine kinase-MB (CK-MB), and the time course of their appearance in plasma correlated with that of neutrophil count and plasma D-dimer. In individual patients, peak and mean sICAM-1 levels correlated respectively with plasma D dimer concentrations and monocyte count, but no correlation were found when their time courses were analyzed. Eight hours after symptom onset, the mean plasma sE selectin levels were higher in patients with AMI than in those with stable angina, whereas no significant differences were found in mean plasma sICAM-1 levels between the two groups at every time analyzed. In the acute phase of MI (a) sE-selectin and sICAM-1 levels increase during the first 8 h and subsequently decrease; (b) the increase in sE-selectin probably reflects activation of endothelial cells, correlates with other inflammatory and coagulation parameters, and is inversely related to the degree of myocardial damage; and (c) sICAM-1 plasma levels do not represent a good marker of "cell activation" because they reflect activation of different cells and may be affected by different conditions. PMID- 9335405 TI - ACE inhibitor effects on platelet function in stages I-II hypertension. AB - Angiotensin II enhances platelet aggregation through activation of the G protein linked pathway present in platelets. Studies of several angiotensin-converting enzyme (ACE) inhibitors have demonstrated marked differences on platelets. Therefore this prospective, randomized, double-blind, crossover study compared the ex vivo effects of equivalent antihypertensive doses of captopril, enalapril, and fosinopril on platelet aggregation and thromboxane B2 (TxB2) formation in subjects with stage I-II essential hypertension. Nineteen male subjects with a baseline mean seated blood pressure of 141 +/- 3/100 +/- 1 mm Hg were enrolled. The decline in mean arterial pressure after 4 weeks of stable dosing was 10 +/- 1, 12 +/- 1, and 11 +/- 1 mm Hg for captopril, enalapril, and fosinopril, respectively (p = NS). There was no significant change in adenosine diphosphate (ADP)-, epinephrine-, or thrombin-stimulated platelet aggregation from baseline or between ACE inhibitors. Compared with baseline, fosinopril decreased TxB2 concentrations 27.5-67.6% with all stimuli after 1 and 5 min. Captopril also decreased TxB2 formation, but this effect was stimulus and time dependent. Enalapril consistently increased TxB2 concentrations, independent of stimuli or time. We conclude that different ACE inhibitors have distinct effects on platelet TxB2 formation without significant effects on platelet aggregation. Fosinopril may be a direct antagonist ofTxA2 synthase, suggesting benefit in syndromes of platelet activation or vascular occlusion. PMID- 9335406 TI - Inhibition of carnitine synthesis protects against left ventricular dysfunction in rats with myocardial ischemia. AB - During myocardial ischemia, inhibition of the carnitine-mediated transportation of fatty acid may be beneficial because it facilitates glucose utilization and prevents an accumulation of fatty acid metabolites. We orally administered 3 (2,2,2-trimethyl hydrazinium) propionate (MET), an inhibitor of carnitine synthesis, for 20 days to rats. Then we evaluated left ventricular (LV) function during brief ischemia by using a buffer-perfused isovolumic heart model. After 15 min of reoxygenation after the transient ischemia, LV peak systolic pressure (PSP) almost completely returned to the baseline level in rats given MET (96 +/- 4%), whereas it was only partially (77 +/- 16%) recovered in the placebo-treated rats. We induced myocardial infarction in other rats by ligating the left anterior descending coronary artery. Then the animals were given MET for 20 days, and LV function was compared. In the placebo-treated rats (with myocardial infarction, but without drug treatment), LVPSP was lower than that in the sham group [108 +/- 19 (n = 10) vs. 136 +/- 15 mm Hg (n = 13); p < 0.05], and the time constant (T) of LV pressure decay was elongated (36 +/- 4 vs. 30 +/- 7 ms; p < 0.05). In MET-treated groups, however, neither PSP nor T differed from those in the sham group. In conclusion, inhibition of the carnitine-mediated transportation of fatty acid by MET protected against left ventricular dysfunction in acute and chronic myocardial ischemia. PMID- 9335407 TI - Hemodynamic responses to endothelin-1 and endothelin antagonists microinjected into the nucleus tractus solitarius in rats. AB - The role of endothelin-1 (ET-1) within the nucleus tractus solitarius (NTS) in central cardiovascular control was investigated by local microinjections of ET-1 and ET-receptor antagonists. In urethane-anesthetized Sprague-Dawley rats, a unilateral microinjection of ET-1 (1.0, 3.3, and 10.0 pmol) into the NTS significantly increased arterial pressure, left ventricular systolic pressure, and dP/dt(max) in a dose-dependent manner, and slightly decreased heart rate in a dose-independent manner. The pressor effect lasted >90 min. In normotensive rats, neither PD147953, a selective ETA-receptor antagonist, nor PD142893, a mixed ETA- and ETB-receptor antagonist, microinjected into the NTS elicited any changes in arterial pressure or heart rate. The pressor and bradycardic effects evoked by microinjection of ET-1 into the NTS could be blocked by local pretreatment with PD147953 and completely eliminated by intravenous pretreatment with the ganglionic blocker hexamethonium. The arterial baroreflex sensitivity was almost totally suppressed by microinjection of ET-1 (3.3 pmol) in alpha-chloralose anesthetized Sprague-Dawley rats. A similar pattern of changes in the hemodynamic variables was elicited by microinjection of ET-1 (3.3 pmol) into the NTS in spontaneously hypertensive rats (SHRs) compared with Wistar-Kyoto (WKY) rats. In SHRs, microinjection of PD142893 did not elicit any changes in arterial pressure or heart rate. These results suggest that ET-1 modulates reflex control of hemodynamics by activation of autonomic nerve via ETA receptors in the NTS, and that the responsiveness of SHRs to ET-1 or PD142893 is similar to that of WKY rats. PMID- 9335408 TI - Short-term estrogen augments both nitric oxide-mediated and non-nitric oxide mediated endothelium-dependent forearm vasodilation in postmenopausal women. AB - Estrogen is known to improve in the short term the impaired endothelium-dependent vasodilating responses in postmenopausal women, which may account in part for the beneficial cardiovascular effects of the female hormone. Endothelium-dependent vasodilation is achieved by combined effects of endothelium-derived prostacyclin, nitric oxide (NO), and hyperpolarizing factor. In this study, we investigated our hypothesis that short-term estrogen improves both NO-mediated and non-NO-mediated endothelium-dependent vasodilation in postmenopausal women. The study included 12 postmenopausal women (aged 64 +/- 3 years). The forearm blood flow was measured by strain-gauge plethysmography. The forearm vascular responses to the endothelium-dependent vasodilators, acetylcholine and substance P, were examined before and after intravenous administration of conjugated estrogen and subsequently after intraarterial infusion of NG-monomethyl-L-arginine (L-NMMA), an inhibitor of NO synthesis. Short-term estrogen augmented the forearm vasodilating responses to both acetylcholine and substance P. The treatment with L-NMMA almost abolished the augmented response to acetylcholine but did not affect that to substance P. The forearm vascular response to sodium nitroprusside was unchanged by the estrogen administration. These results indicate that estrogen augments (in the short-term) both NO-mediated and non-NO-mediated endothelium-dependent forearm vasodilation in postmenopausal women. Thus the beneficial effect of estrogen on endothelial vasodilator function appears to extend to non-NO-dependent mechanism(s). PMID- 9335409 TI - Multiple modulations of action potential duration by different calcium channel blocking agents in guinea pig ventricular myocytes. AB - Effects of extracellular applications of different types of Ca2+ channel blocking agents (Mn2+, verapamil, and nisoldipine) on action-potential duration and membrane currents were studied by the whole-cell patch-clamp technique in guinea pig ventricular myocytes. Low concentrations of Mn2+ (1 mM) and verapamil (1 microM) prolonged action-potential duration at 90% repolarization (APD90) with a suppressed plateau phase. Increases in Mn2+ (5 mM) and verapamil (5 microM) shortened APD90 with a further depression of the plateau. Nisoldipine (0.2-1 microM) shortened APD90 without lengthening it. Applications of Mn2+ and verapamil suppressed amplitudes of the L-type Ca2+ current (ICa), the delayed outward K+ current (IK), and the inward rectifier K+ current (IK1). Furthermore, the ratios of ICa:IK inhibition were similar by low and high concentrations of Mn2+ and verapamil. Nisoldipine selectively suppressed ICa without effect on IK and IK1. A low concentration (1 mM) of Mn2+ not only decreased the peak amplitude of ICa but also delayed its decay time course, which caused an increase in late ICa amplitude at the end of a 200-ms depolarizing pulse. Both verapamil and nisoldipine suppressed peak ICa without affecting its decay. Whereas Mn2+ suppressed IBa without changing its decay time course, verapamil and nisoldipine speeded up the IBa decay with suppressed amplitude of IBa. We conclude that different types of Ca2+ channel blocking agents (Mn2+, verapamil, and nisoldipine) diversely modulate APD because of their multiple modes of actions on ICa and IK. PMID- 9335410 TI - Combined therapy with benazepril and amlodipine in the treatment of hypertension inadequately controlled by an ACE inhibitor alone. AB - In a multicenter, randomized, double-blind, placebo-controlled study, we evaluated the efficacy and tolerability of the combination of benazepril, 10 mg, and amlodipine, 2.5 or 5 mg once daily, compared with benazepril, 10 mg, monotherapy in patients with hypertension inadequately controlled with angiotensin-converting enzyme (ACE)-inhibitor monotherapy. After a 2-week placebo and 4-week single-blind benazepril, 10 mg once daily, run-in period, 448 patients, 213 men and 235 women, aged 24-73 years (mean, 55 years), with mean diastolic blood pressure (DBP) > or =95 and < or =120 mm Hg at the end of the benazepril run-in period, were randomized to receive one of the following treatments once daily for 8 weeks: (a) benazepril, 10 mg, plus placebo (BZ10); (b) benazepril, 10 mg, plus amlodipine, 2.5 mg (BZ10/AML2.5); or (c) benazepril, 10 mg, plus amlodipine, 5 mg (BZ10/AML5). Before the patients were admitted to the trial, at the end of the placebo run-in and the benazepril run-in period and at the end of weeks 4 and 8 of the treatment period, sitting and standing blood pressure (BP), heart rate (HR), and body weight were measured 22-26 h after the intake of the trial medication. Both BZ10/AML2.5 and BZ10/AML5 combinations showed better antihypertensive activity than did BZ10 monotherapy at the terminal visit as demonstrated by (a) the 24-h postdosing sitting and standing systolic BP (SBP) and DBP values, which were statistically lower with combination therapy than with BZ10; (b) the success rate, which was statistically higher with both the combinations (69.2% in the BZ10/AML2.5 and 65.8% in the BZ10/AML5 group) compared with the BZ10 group (40.5%). The tolerability was good in the three treatment groups. No significant abnormal laboratory data were detected. There was no difference in efficacy and safety/tolerability between the BZ10/AML2.5 and BZ10/AML5 groups. PMID- 9335411 TI - Physiological role of Ca2+-permeable nonselective cation channel in endothelin-1 induced contraction of rabbit aorta. AB - We previously showed a role for a nonselective cation channel (NSCC) in the ETA dependent action of endothelin-1 in mouse fibroblast and rabbit aortic smooth muscle cell. To clarify the physiological significance of NSCCs in endothelin-1 (ET-1)-induced vasocontraction, we examined the effects of NSCC blockers such as mefenamic acid and SK&F 96365 on the contractions of deendothelialized rabbit aortic rings induced by a low (10[-10] M) or high (10[-8] M) concentration of ET 1. Mefenamic acid (< or =10[-3] M) had little effect on the contraction induced by 45 x 10(-3) M K+ or 1 x 10(-6) M Bay K-8644 in combination with 15 x 10(-3) M K+, indicating that it does not affect voltage-operated calcium channels (VOCs) and contractile mechanisms. The contraction by a low concentration of ET-1 was abolished after removal of extracellular Ca2+, but it was reduced only to 50% by a maximally effective concentration (10[-5] M) of nifedipine, an inhibitor of L type VOCs (L-VOC). Mefenamic acid and SK&F 96365 inhibited the ET-1-induced contraction with 50% inhibitory concentration (IC50) values of 10(-4) M and 2 x 10(-5) M, respectively, and abolished it at 10(-3) M and 10(-4) M. By contrast, nifedipine, mefenamic acid, or SK&F 96365 had little effect on the contraction by a high concentration of ET-1. The contraction induced by a low or high concentration of ET-1 was abolished by an ETA antagonist, BQ-123, but not by an ETB antagonist, BQ-788. These results demonstrate that the contraction induced by ET-1 is totally mediated exclusively by ETA, but that Ca2+ entry through NSCCs in addition to L-VOCs plays an important role in contractions induced by low concentrations of ET-1, whereas it plays only a minor role in contractions induced by high concentrations of ET-1. PMID- 9335412 TI - Short-term lisinopril treatment in old rats worsens impairment of angiotensin induced reflex bradycardia. AB - To determine how short-term treatment with an angiotensin-converting enzyme (ACE) inhibitor affects drug-induced reflex bradycardia at different ages in conscious rats, we compared the magnitude of drug-induced reflex bradycardia before and after injecting bolus intravenous doses of lisinopril, 1 mg/100 g, in male Sprague-Dawley rats aged 4 (young) or 19 (old) months. Anesthetic artifacts were avoided by recording all drug-induced cardiovascular responses from femoral arterial cannulas implanted 1 week earlier. For eliciting reflex bradycardia, blood pressure was increased by graded intravenous infusion of angiotensin or phenylephrine. Impairment of reflex bradycardia in old rats occurred only during pressor responses to angiotensin but not when blood pressure was equally increased with phenylephrine. Subsequent administration of lisinopril affected neither pressor and reflex bradycardic responses to phenylephrine nor pressor responses to angiotensin. However, contrary to the baroreflex enhancement described previously by others, the reflex bradycardia induced by angiotensin was reduced by lisinopril treatment but only in old and not in young rats. Thus our results indicate that whereas angiotensin-induced reflex bradycardia was already impaired in old rats before lisinopril was given, it was reduced further after short-term lisinopril treatment. PMID- 9335413 TI - Beta-amyloid-induced coronary artery vasoactivity and endothelial damage. AB - Amyloid beta-peptide (A beta) deposition has been associated with coronary heart disease and neurodegenerative diseases. A link between A beta and free radical generation has been explored in neuronal tissue. We report here on the effect of A beta on pressurized segments of coronary resistance arteries and the role of free radicals. A small oscillatory response to A beta (10[-6] M) that consisted of a relaxation followed by constriction and a return to the basal diameter was observed in all vessels. The thromboxane A2 analog U46619 produced a significantly greater constriction compared with the response before treatment with A beta. The presence of the antioxidant enzyme superoxide dismutase (SOD) reduced both the response to A beta alone and the enhanced response to U46619. Vasodilation responses to acetylcholine (10[-9]-10[-5] M) were virtually eliminated at all concentrations by A beta. We confirmed endothelial cell damage by A beta with electron microscopy. The results suggest that A beta deposition in coronary resistance arteries causes endothelial damage that is mediated through superoxide radicals. PMID- 9335414 TI - Relations between fasting serum insulin, glucose, and dihydroepiandrosterone sulfate concentrations in obese patients with hypertension: short-term effects of antihypertensive drugs. AB - A randomized, single-blind, placebo-controlled study was conducted in 82 obese patients with mild to moderate essential hypertension, to determine the incidence of hyperinsulinemia, the relations between fasting insulin and dihydroepiandrosterone-sulfate (DHEA-S) levels, and the short-term effects of antihypertensives on DHEA-S and insulin serum concentrations. Increased insulin/glucose ratios (IGR) suggestive of insulin resistance were found in half of our patients. Hyperinsulinemic and normoinsulinemic obese patients with hypertension had comparable fasting glucose and DHEA-S concentrations and comparable blood pressure (BP) levels. Thus no relations were found between fasting insulin and DHEA-S levels. Fasting hyperinsulinemia was found in only half of the obese subjects with hypertension, suggesting that not all obese patients with hypertension are at the same high cardiovascular risk. Short-term treatment with captopril, prazosin, verapamil, atenolol, or hydrochlorothiazide (HCTZ) reduced BP; greater BP reduction was observed with drugs with vasodilatory effects. Captopril, prazosin, and verapamil reduced fasting insulin levels, whereas atenolol and hydrochlorothiazide did not. The former drugs reduced fasting insulin levels that were either within normal limits or in the hyperinsulinemic range. None of the drug treatments produced significant increases in serum DHEA-S concentrations, although some of them considerably reduced fasting insulin levels. No relations between insulin and DHEA-S levels were observed either at baseline or at the end of the antihypertensive treatment. The BP reduction resulting from the peripheral vasodilation may explain the insulin-reducing action of captopril, verapamil, and prazosin. These results further emphasize the large heterogeneity present in the pathophysiologic mechanisms operating in obesity and hypertension. PMID- 9335415 TI - Activation of nuclear factor-kappaB in cultured endothelial cells by increased glucose concentration: prevention by calphostin C. AB - Nuclear factor kappaB (NFkappaB) plays a pivotal role in early gene responses by promoting messenger RNA (mRNA) synthesis for various cell-adhesion molecules and inducible nitric oxide synthase. In this study, we examined whether increases in glucose concentration enhance NFkappaB expression in nuclear fractions of endothelial cells by using electrophoretic mobility shift assay. Bovine aortic endothelial cells (BAECs) were incubated in media containing 5.5-35 mM glucose. NFkappaB activity was increased as early as 1 h (peak activation at 2-4 h) after incubation with 35 mM glucose compared with 5.5 mM. Similar increases at 2 h of incubation were observed by using 25 but not 15 mM glucose. Glucose-induced NFkappaB activation was blocked by inhibiting nuclear translocation by using a peptide (SN-50) containing the nuclear-localization sequence of NFkappaB p50 linked to a membrane-permeable motif of the sequence for Kaposi fibroblast growth factor. Co-incubation with a selective protein kinase C (PKC) inhibitor, calphostin C, produced a concentration-dependent inhibition of glucose-induced NFkappaB activation. Thus NFkappaB activation is an early event in response to elevations in glucose, which may elicit multiple pathways contributing to the origin of hyperglycemia- or diabetes-induced endothelial cell injury. PMID- 9335416 TI - Angiotensin II-receptor blockade further reduces afterload safely in patients maximally treated with angiotensin-converting enzyme inhibitors for heart failure. AB - Combined therapy with an angiotensin-II type I receptor (AT1) antagonist and an angiotensin-converting enzyme (ACE) inhibitor results in more complete suppression of the renin-angiotensin system. Accordingly, the blood-pressure response and safety of combining AT1-receptor blockade with losartan for ACE inhibition were evaluated in patients with congestive heart failure who were already treated with maximally recommended or tolerated doses of an ACE inhibitor. Forty-three patients with symptomatic congestive heart failure were evaluated biweekly for 1 month before addition of losartan and weekly during administration of losartan at a daily dose of 25 mg for the first week and 50 mg for the second week. Systolic blood pressure, which remained unchanged before addition of losartan, decreased from 122 +/- 18 mm Hg to 112 +/- 17 and 107 +/- 17 mm Hg (p < 0.001) after 1 week of 25 mg and 1 week of 50 mg losartan, respectively. Diastolic blood pressure also significantly decreased. The decreases in blood pressure were well tolerated by all patients, even by those in whom symptomatic hypotension developed during uptitration of ACE inhibition. Serum potassium and sodium and parameters of renal function remained unchanged. Combining AT1-receptor blockade with losartan to maximally recommended or tolerated ACE inhibition appears safe and leads to further vasodilatation in symptomatic patients with congestive heart failure. PMID- 9335417 TI - Cyclic GMP decreases cardiac myocyte oxygen consumption to a greater extent under conditions of increased metabolism. AB - We tested the hypothesis that the negative effects of intracellular guanosine 3',5'-cyclic monophosphate (cyclic GMP) were more profound on cardiac myocyte oxygen consumption (VO2) during increased metabolism of the myocytes. The steady state VO2 of a suspension of single myocytes isolated from hearts of New Zealand White rabbits was measured in a glass chamber by using a Clark-type oxygen electrode, and cyclic GMP was determined by using a radioimmunoassay. The cellular cyclic GMP levels were increased either by adding 3-morpholino sydnonimine (SIN-1), a guanylate cyclase stimulator, or zaprinast (ZAP), a cyclic GMP-phosphodiesterase inhibitor, at various doses. In 0.5 mM Ca2+ medium, average VO2 was 123 +/- 8 nl/min/100,000 cells, and cyclic GMP was 35.4 +/- 9.3 fmol/100,000 cells, and these increased significantly to 182 +/- 9 nl/min/100,000 cells and 78.2 +/- 7.3 fmol/100,000 cells in 2.0 mM Ca2+. There were dose dependent responses of the VO2 and cellular cyclic GMP levels in responding to both SIN-1 and ZAP. An inverse relation between cellular cyclic GMP level and VO2 existed in the myocytes. The regression equations for the four treatments were log(VO2) = -0.002[cyclic GMP] + 2.19, r = 0.96 for SIN-1 in low (0.5 mM) Ca2+; log(VO2) = 0.005[cyclic GMP] + 1.80, r = 0.38 for ZAP in low Ca2+; log(VO2) = 0.001 [cyclic GMP] + 2.24, r = 0.82 for SIN-1 in high (2.0 mM) Ca2+; and log(VO2) = -0.004[cyclic GMP] + 2.56, r = 0.93 for ZAP in high Ca2+. The slope of ZAP regression line was significantly more negative than that of SIN-1 with high calcium. At any given level of cyclic GMP, ZAP decreased the VO2 to a greater extent than did SIN-1 although the latter caused the maximal increase in cyclic GMP level. The reduction in VO2 caused by a corresponding increase in cellular cyclic GMP was greater in myocytes incubated with high-Ca2+ medium. PMID- 9335418 TI - Natural variability in the number of dendritic segments: model-based inferences about branching during neurite outgrowth. AB - A study was made of the possible basis for naturally occurring variations in the number of segments in individual dendritic trees. Distributions of the number of terminal segments have been studied in dendrites from rat, cat, and frog motoneurons, basal dendrites from rat visual cortex pyramidal and non-pyramidal neurons, in rat cerebellar Purkinje cell dendritic trees, and in human hippocampal dentate granule cells. By means of a mathematical model for dendritic branching, it was shown that the variation in the number of dendritic segments can be accounted for by assuming that new branches during neurite outgrowth are formed randomly at terminal segments. The observed terminal segment number distributions could be closely approximated by additionally assuming that branching probabilities decline with increasing number of terminal segments in growing dendrites. The pyramidal neuron group differed significantly from the other neuron groups in such a way as to suggest that this decline is stronger than in the dendrites of other types of neurons. By using literature data on the mean number of terminal segments in rat cerebellar Purkinje cells, measured at different times during early development, an estimate could be obtained of the time-course of the branching probabilities. The branching probability of a terminal segment was found to be in the order of 0.002 per hour in the first 4 weeks postnatal with a 5-fold transient increase in the second week. PMID- 9335419 TI - Calretinin-like immunoreactivity in mormyrid and gymnarchid electrosensory and electromotor systems. AB - Calretinin-like immunoreactivity was examined in the electrosensory and electromotor systems of the two families of mormyriform electric fish. Mormyrid fish showed the strongest immunoreactivity in the knollenorgan electroreceptor pathway; in the nucleus of the electrosensory lateral line lobe (ELL) and the big cells of the nucleus exterolateralis pars anterior. Mormyromast and ampullary zones of the ELL showed calretinin-like immunoreactivity in the ganglion, granule, and intermediate cell and fiber layers. Mormyromast zones additionally showed labeling of apical dendrites and commissural cells, but the ampullary zone did not. In the electromotor system, two nuclei in the corollary discharge pathway showed labeling: in the paratrigeminal command-associated nucleus and the juxtalobar nucleus. Gymnarchus niloticus (Gymnarchidae) showed strongest calretinin-like immunoreactivity in part of the phase-coding pathway; in S-type electroreceptor afferents. Zones of the ELL not receiving phase-coder input had weak labeling. The electromotor system showed labeling in the lateral relay nucleus and less strongly in the medullary relay nucleus, but none in the pacemaker. The concentration of calcium-binding proteins in mormyrid and gymnarchid time-coding electrosensory pathways is consistent with the hypothesis that they play a role in preserving temporal information across synapses. Cell types that encode temporal characteristics of stimuli in precise spike times have high levels of calcium-binding proteins, but cells that re-code temporal information into presence or magnitude of activity have low levels. Some cell types in the electromotor pathways and early in the time-coding electrosensory pathways do not follow this hypothesis, and therefore preserve temporal information using a mechanism independent of calcium-binding proteins. In particular, electromotor systems may use extensive electrotonic coupling within nuclei to ensure precise timing. PMID- 9335420 TI - Optically imaged maps of orientation preference in primary visual cortex of cats and ferrets. AB - Feature maps in the cerebral cortex constitute orderly representations of response features created within the cortex; an example is the mapping of orientation-selective neurons in visual cortex. We have compared the properties of orientation maps in area 17 of cats and ferrets, obtained by optical imaging of intrinsic signals. Orientation maps in both species contain a quasi-periodic distribution of iso-orientation domains that are organized into a lattice of pinwheels. However, the spatial density of orientation domains and of pinwheels in ferret area 17 is nearly twice that in cat area 17. The ferret map also contains more discontinuities, or fractures, where orientation changes abruptly. The size of orientation domains scales with interdomain spacing, so that the ratio of the two is approximately the same in both species. Consistent with this finding, the orientation tuning width of individual pixels is similar in the two. The magnitude of orientation preference, however, is much lower in ferret compared to cat. The greater incidence of fractures in ferret appears to be due to proportionately greater overlap between domains of different orientations, particularly along fracture lines that link pinwheel centers. We hypothesize that a key determinant of orientation maps, the relationship between orientation domain size and spacing, expresses an anatomical link between sizes of thalamocortical arbors and horizontal intracortical connections in area 17. PMID- 9335421 TI - Identification of transient microglial cell colonies in the forebrain white matter of developing rats. AB - Herein, we describe the existence of distinct colonies of transient microglial cells that reside in well-defined zones of the forebrain white matter. Rats, aged at postnatal day (P) 0, P2, P5, P7, P10, P15 or adult, were anaesthetised with halothane gas, and various neural centres were injected unilaterally with the tracer biotinylated Dextran. The neural centres injected were cingulate or sensorimotor cortices, ventral nuclei of the dorsal thalamus, and the pontine reticular formation of the brainstem. Rats were allowed to survive to various stages, from 4 hours to 21 days, after the injection. They were then anaesthetised with sodium pentobarbitone, and their brains were aldehyde-fixed and processed by using standard methods. The following is a description of what is seen after injections at P0, P2, P5, P7, P10; we saw no labelled cells (described below) in the rats injected at P15 or adult. From 2 to 21 days after an injection of dextran into the above-mentioned centres, labelled microglial cell colonies, identified by using double-labelling with anti-OX-6 or Griffonia simplicifolia (Bandeiraea; isolectin B4), were seen in small isolated zones in the forebrain white matter. These colonies were in the corpus callosum, the dorsal and ventral regions of the external capsule, and the internal capsule. A striking feature of these labelled microglial cell colonies was that they were seen on both sides of the brain. Thus, regardless of the location of the injection site in either the cortex, thalamus, or brainstem, the same microglial cell colonies were labelled with dextran in the forebrain white matter. After injections of different coloured fluorescent dextrans into the cortex and into the brainstem of the same animal, many double-labelled cells in each of the colonies were seen. From our short-term survival cases (4 hours to 1 day), a rather strict sequence or progression of labelling of the colonies across the white matter from the injection site was seen; in general, the microglial cell colonies closest to the injection site became labelled well before (about a day) those further away. These results lead us to suggest that the microglial cells in each colony become labelled after a slow diffusion of the tracer through the extracellular space from the injection site. PMID- 9335422 TI - Comparison of the regional expression of nicotinic acetylcholine receptor alpha7 mRNA and [125I]-alpha-bungarotoxin binding in human postmortem brain. AB - Neuronal nicotinic acetylcholine receptors are expressed in the human central nervous system. A specific subtype of this receptor family, the alpha7 nicotinic acetylcholine receptor, is thought to be the principal alpha-bungarotoxin (alphaBTX)-binding protein in mammalian brain. Although the expression of this receptor subtype has been characterized in rat, no study has specifically compared the expression of both the alpha7 gene and the localization of BTX binding sites in human brain. Expression of alpha7 mRNA and receptor protein in human postmortem brain tissue was examined by in situ hybridization and [125I] alpha-bungarotoxin autoradiography, respectively, with particular emphasis on regions associated with sensory processing. Regions with high levels of both alpha7 gene expression and [125I]-alphaBTX binding include the nucleus reticularis of the thalamus, the lateral and medial geniculate bodies, the basilar pontine nucleus, the horizontal limb of the diagonal band of Broca, the nucleus basalis of Meynert, and the inferior olivary nucleus. High-to-moderate levels of alpha7 probe hybridization were also seen in the hippocampus and the cerebral cortex; however, there was a reduced or variable degree of [125I] alphaBTX binding in these regions compared with the level of probe hybridization. In most brain regions, [125I]-alphaBTX binding was localized to neuronal cell bodies similar in morphology to those that exhibited alpha7 hybridization, suggesting that the high-affinity [125I]-alphaBTX binding sites in the human brain are likely to be principally composed of alpha7 receptor subtypes. PMID- 9335423 TI - Distribution of radial glia in the developing telencephalon of chicks. AB - Radial glia are known to have a sparse and uneven distribution in the telencephalon of adult birds. The present study utilizes antibodies against vimentin to reveal a more extensive, and more clearly radial, set of radial glia in the chicken telencephalon during the first half of embryogenesis. This initially extensive radial glial fiber system becomes distorted and reduced between 10 and 14 days of incubation. This reduction coincides with the cytoarchitectural differentiation of the telencephalon into its major adult subdivisions. Because developing neurons tend to migrate along radial glial fibers in both birds and mammals, a topological projection of these major subdivisions onto the embryonic ventricular zone along the radial glial fibers suggests hypotheses about lineage relationships that can be tested by subsequent experimental methods. This analysis suggests that the major components of the avian dorsal ventricular ridge, i.e., the ventral hyperstriatum, the neostriatum with its various subdivisions, part of the archistriatum, and probably also the piriform cortex, all derive from overlapping portions of the lateral pallial ventricular zone. Staining with antibodies against neurofilament suggests that this developmental parcellation of the lateral pallial complex is associated with the development of neuronal fiber systems. PMID- 9335424 TI - Naturally occurring neuron death during postnatal development of the gerbil ventral cochlear nucleus begins at the onset of hearing. AB - Postnatal development of the gerbil ventral cochlear nucleus (VCN) was studied quantitatively under the light microscope in Nissl-stained serial sections at postnatal day 0 (P0), P5, P7, P10, P12, P15, and P140. VCN boundaries were unambiguous at all ages, and nucleus volume was calculated planimetrically for all groups. Measurements of neuron soma cross-sectional area and number were made in all groups except P0. Both VCN volume and soma area doubled between P5 and P10. Although somatic growth did not continue beyond P10, VCN volume increased a further 57% between P15 and P140. Neuron number did not change significantly between P5 and P10, averaging approximately 36,000 neurons. Between P10 and P12, neuron number decreased significantly by 22%, with no further change thereafter. Our data show that, following significant postnatal growth in the gerbil VCN, a brief period of naturally occurring neuron death begins at the onset of hearing. PMID- 9335425 TI - A confocal and electron microscopic study of contacts between 5-HT fibres and feline dorsal horn interneurons in pathways from muscle afferents. AB - Morphological substrates of actions of serotonin upon dorsal horn interneurons with input from group II muscle afferents were investigated by using two experimental approaches. Twelve interneurons were intracellularly labelled with rhodamine-dextran, and serotoninergic fibres were identified by immunofluorescence. Appositions between the serotoninergic axons and these interneurons were examined with a dual-channel confocal microscope. A further four interneurons were intracellularly labelled with horseradish peroxidase, and serotoninergic axons were identified by immunocytochemistry; these neurons were prepared for combined light and electron microscopy. Confocal microscopy revealed serotoninergic varicosities in apposition to both cell bodies and dendrites. Similar total numbers of appositions were found on the soma, and on dendrites within 100 microm from the soma, on the most completely labelled neurons. The number of appositions on 100-microm segments of dendrites decreased with increasing distances from the soma (from 14.6 within 100 microm, to 3.8 and 2.4 at 100-300 microm, and more than 300 microm distances, respectively). Electron microscopic analysis of two neurons revealed that few of the apparent contacts on cell bodies were synaptic, but, in contrast, many varicosities apposed to proximal dendrites formed synapses. The evidence suggests that serotonin may have more powerful synaptic effects upon the dendrites of this class of dorsal horn interneurons than on their cell bodies. PMID- 9335426 TI - Vsx-1 and Vsx-2: two Chx10-like homeobox genes expressed in overlapping domains in the adult goldfish retina. AB - The genetic linkages of the murine ocular retardation mutation with the Chx10 gene and the murine small eye mutation with the Pax-6 gene has demonstrated the importance of Paired class homeobox genes in the development of the mammalian retina. Previously, we identified a Paired-class homeobox gene, Vsx-1, whose expression in the adult goldfish retina is restricted to the inner nuclear layer (INL) and to postmitotic, differentiating progenitor cells in the growth zone at the retinal peripheral margin, where neurogenesis continues throughout life. Here, we report the molecular cloning and expression pattern of a new Paired class homeobox gene, Vsx-2, in the adult goldfish retina. Like Vsx-1, Vsx-2 expression is highly restricted to the retina in the adult goldfish and overlaps with Vsx-1 expression in the mature INL. At the peripheral margin, Vsx-2 is expressed in mitotically active neuronal progenitors and is downregulated as these cells become postmitotic and begin to differentiate. Comparison of the amino acid sequences of Vsx-2, Vsx-1, Chx10, and C. elegans ceh-10 reveal a conserved homeodomain and a unique domain termed the CVC domain. The similarities of the Vsx-2, Vsx-1, and Chx10 expression patterns suggest that genes containing the CVC domain have conserved functions during retinal development in vertebrates. PMID- 9335427 TI - Two distinct populations of tectal neurons have unique connections within the retinotectorotundal pathway of the pigeon (Columba livia). AB - The tectofugal pathway is a massive ascending polysynaptic pathway from the tectum to the thalamus and then to the telencephalon. In birds, the initial component of this pathway is known as the tectorotundal pathway; in mammals, it is known as the tectopulvinar pathway. The avian tectorotundal pathway is highly developed; thus, it provides a particularly appropriate model for exploring the fundamental properties of this system in all amniotes. To further define the connectivity of the tectorotundal projections of the tectofugal pathway, we injected cholera toxin B fragment into various rotundal divisions, the tectobulbar projection, and the ventral supraoptic decussation of the pigeon. We found intense bilateral retrograde labeling of neurons that stratified within layer 13 and, in certain cases, granular staining in layer 5b of the optic tectum. Based on these results, we propose that there are two distinct types of layer 13 neurons that project to the rotundus: 1) type I neurons, which are found in the outer sublamina of layer 13 (closer to layer 12) and which project to the anterior and centralis rotundal divisions, and 2) type II neurons, which are found in the inner sublamina of layer 13 (closer to layer 14) and which project to the posterior and triangularis rotundal divisions. Only the labeling of type I neurons produced the granular dendritic staining in layer 5b. An additional type of tectal neuron was also found that projected to the tectobulbar system. We then injected Phaseolus vulgaris-leucoagglutinin in the optic tract and found that the retinal axons terminating within tectal layer 5b formed narrow radial arbors (7 10 microm in diameter) that were confined to layer 5b. Based on these results, we propose that these axons are derived from a population of small retinal ganglion cells (4.5-6.0 microm in diameter) that terminate on the distal dendrites of type I neurons. This study strongly indicated the presence of a major bilateral oligosynaptic retinotectorotundal pathway arising from small retinal ganglion cells projecting to the rotundus with only a single intervening tectal neuron, the proposed type I neuron. We suggest that a similar organization of retinotectopulvinar connections exist in reptiles and in many mammals. PMID- 9335428 TI - The double-stranded RNA-dependent protein kinase PKR: structure and function. AB - This review describes the structure and function of the interferon (IFN) inducible, double-stranded RNA-activated protein kinase PKR. This protein kinase has been studied extensively in recent years, and a large body of evidence has accumulated concerning its expression, interaction with regulatory RNA and protein molecules, and modes of activation and inhibition. PKR has been shown to play a variety of important roles in the regulation of translation, transcription, and signal transduction pathways through its ability to phosphorylate protein synthesis initiation factor eIF2, I-kappaB (the inhibitor of NF-kappaB), and other substrates. Expression studies involving both the wild type protein and dominant negative mutants of PKR have established roles for the enzyme in the antiviral effects of IFNs, in the responses of uninfected cells to physiologic stresses, and in cell growth regulation. The possibility that PKR may function as a tumor suppressor and inducer of apoptosis suggests that this IFN regulated protein kinase may be of central importance to the control of cell proliferation and transformation. PMID- 9335429 TI - Long-term interferon-alpha therapy does not affect sex hormones in males with chronic hepatitis C. AB - The effects of interferon-alpha (IFN-alpha), given at a dosage of 6 MU thrice weekly for 12 months, on gonadal function were investigated in 18 males affected by chronic hepatitis C. Periodically, all patients were clinically monitored and questioned about sexual function. Gonadotropin and serum androgen concentrations (follicle-stimulating hormone, luteinizing hormone, total testosterone, free testosterone, androstenedione, dehydroepiandrosterone, dehydroepiandrosterone sulfate, and sex hormone binding globulin) were tested every 3 months. Ten of 18 patients (55%) responded to IFN-alpha therapy. Serum total testosterone and sex hormone binding globulin values decreased slightly at the third month of treatment, then returned to baseline values. Serum free testosterone and other sex hormones remained essentially unchanged during IFN-alpha therapy. Four patients (22.2%) complained of sexual dysfunction (impaired libido, erectile failure, and impaired ejaculation), which was unrelated to any significant hormonal change and resolved after IFN therapy was stopped. Serum sex hormones values did not differ between responders and nonresponders to IFN-alpha. This study indicates that 12 months treatment with 6 MU of IFN-alpha thrice weekly does not significantly affect gonadal function in men with chronic hepatitis C. The sexual dysfunction observed could be ascribed to such other side effects of IFN as asthenia, fatigue, or anxiety, or it could have a psychologic basis. PMID- 9335430 TI - Effects of varying lengths of double-stranded RNA on binding and activation of 2' 5'-oligoadenylate synthetase. AB - We investigated the effects of varying the length of the double-stranded (ds) RNA cofactor on activation of 2'-5'-oligoadenylate [2-5 (A)] synthetase. dsRNA of 40, 55, 67, 85, and 110 bp lengths were generated by in vitro transcription of complementary strands and annealing and trimming of the single-stranded overhangs. The dsRNA molecules were radiolabeled by polynucleotide kinase and purified by gel electrophoresis. Their abilities to bind to recombinant mouse 9-2 2-5 (A) synthetase, as monitored by electrophoretic mobility shift assays, were comparable. When used at a saturating concentration of 25 microg/ml, all dsRNA molecules were equally effective in activating the enzyme. At a subsaturating concentration, however, longer RNAs were better activators. At 100 nM concentration, there was a linear relationship between the length of the dsRNA and its ability to activate 2-5 (A) synthetase. PMID- 9335431 TI - Unexpected correlation in the sensitivity of 19 herpes simplex virus strains to types I and II interferons. AB - We compared the sensitivity of 19 herpes simplex virus (HSV) strains to type I (IFN-alpha and IFN-beta) and type II (IFN-gamma) human interferons in cultures of human retinal epithelial (K-1034) and lung (HEL) cells. Their sensitivities proved to be well correlated, even though type I and type II IFN have been reported to have different antiviral actions. The correlation was not because IFN gamma stimulated the formation of IFN-beta, for an antibody that neutralized IFN beta did not reduce its inhibitory effects. Our results show that each HSV strain has a characteristic and similar sensitivity to type I and type II IFN and suggest some common pathway in the mechanism of their antiviral actions. PMID- 9335432 TI - Hypothalamic-pituitary axis remains intact after interferon-alpha treatment in hematologic diseases. AB - Many endocrinologic disturbances have been reported during and after interferon alpha (IFN-alpha) treatment. These disturbances have often been caused by autoantibodies. The aim of this prospective study was to evaluate whether IFN alpha causes hormonal changes and if it is necessary to search for such disturbances routinely. Ten patients with hematologic malignancies were examined before and after 4 months of IFN-alpha treatment. Pituitary function was tested by hypothalamic releasing hormones (thyrotropin-releasing hormone, TRH, growth hormone-releasing hormone, GHRH, gonadotropin-releasing hormone, GnRH). The adrenal glands were tested with the adrenocorticotropin (ACTH) test. The human chorionic gonadotropin (hCG) test was performed on the men (n = 4). The IFN treatment was well tolerated, and no long-term hormonal side effects were found. The testosterone/sex hormone binding globulin (SHBG) index tended to improve. There were no significant differences between the hormone responses before and after IFN-alpha treatment. We conclude that the hypothalamic-pituitary axis remains intact after IFN-alpha treatment. There is no need to follow patients endocrinologically if the patients are not predisposed by autoantibodies. PMID- 9335433 TI - In vivo effects of chicken interferon-gamma during infection with Eimeria. AB - Newly hatched chickens are highly susceptible to infection by opportunistic pathogens during the first 1 or 2 weeks of life. The use of cytokines as therapeutic agents has been studied in animal models as well as in immunosuppressed patients. This approach has become more feasible in livestock animals, in particular poultry, with the recent cloning of cytokine genes and the development of new technologies, such as live delivery vectors. We have recently cloned the gene for chicken interferon-gamma (Ch-IFN-gamma). Poly-HIS-tagged recombinant Ch-IFN-gamma was expressed in Escherichia coli, was purified by Ni chromatography, and was found to be stable at 4 degrees C and an ambient temperature for at least several months and Several weeks, respectively. Ch-IFN gamma was capable of protecting chick fibroblasts from undergoing virus-mediated lysis, induced nitrite secretion from chicken macrophages in vitro, and enhanced MHC class II expression on macrophages. Administration of recombinant Ch-IFN gamma to chickens resulted in enhanced weight gain over a 12-day period. Furthermore, the therapeutic potential of Ch-IFN-gamma was assessed using a coccidial challenge model. Birds were treated with Ch-IFN-gamma or a diluent control and then infected with Eimeria acervulina. Infected birds treated with Ch IFN-gamma showed improved weight gain relative to noninfected birds. The ability of Ch-IFN-gamma to enhance weight gain in the face of coccidial infection makes it an excellent candidate as a therapeutic agent. PMID- 9335434 TI - Both variant forms of interferon-alpha4 gene (IFNA4a and IFNA4b) are present in the human population. AB - Alpha interferons (IFN-alpha) are a class of cytokines with various activities that are used as therapeutic agents for treatment of cancer and viral and immune disorder diseases. At least 13 IFN-alpha genes and 1 IFN-alpha pseudogene have been identified, which are clustered on human chromosome 9. Among the known IFN alpha species, a number of allelic variants have been reported. Two variants of IFN-alpha4 (IFN-alpha4a and IFN-alpha4b) are known, which differ from each other by changes in their coding regions at nucleotide positions 220 and 410 and can be distinguished by selective restriction enzyme analysis. We have developed oligonucleotide primers for specific amplification of IFN-alpha4 gene fragments using the polymerase chain reaction (PCR). Genomic DNA obtained from over 28,000 normal healthy individuals and six human cell lines were used in this study. The resulting PCR products were analyzed by restriction endonuclease digestion and DNA sequencing to identify the presence of variant sequences. The results show that the DNA sequences for both variants of IFN-alpha4 are found in the population in nearly equal proportion. Individuals with either homozygous (e.g., alpha4a/alpha4a or alpha4b/alpha4b) or heterozygous (i.e., alpha4a/alpha4b) IFN alpha4 genes were detected. Among the cell lines, KG-1, EB-3, and HTB-10 cells contain the genes for IFN-alpha4a only, whereas U-937, Namalwa, and Daudi cells contain the genes for both IFN-alpha4a and IFN-alpha4b. PMID- 9335435 TI - Sublethal gamma irradiation increases IL-1alpha, IL-6, and TNF-alpha mRNA levels in murine hematopoietic tissues. AB - The effects of sublethal (7.75 Gy) 60Co gamma radiation exposure on endogenous bone marrow and splenic interleukin-1alpha (IL-1alpha), IL-6, and tumor necrosis factor-alpha (TNF-alpha) mRNA and protein levels were assayed in B6D2F1 female mice. Bone marrow and spleen were harvested from normal and irradiated mice on days 2, 4, 7, 10, and 14 postexposure, and cytokine mRNA levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and Southern blot analysis. IL-1alpha mRNA levels were significantly increased in bone marrow at days 2 and 4 postirradiation and at day 7 in spleen compared with controls. Postirradiation IL-6 mRNA levels showed a significant increase at day 2 in bone marrow and at days 7 and 10 in spleen. TNF-alpha mRNA levels exhibited a significant increase at day 2 postirradiation in bone marrow, but in spleen no difference between control and irradiated samples was observed on any day postirradiation. Interestingly, there were no significant differences in the cytokine protein levels in postirradiation bone marrow, spleen, or serum when compared with normal controls. PMID- 9335436 TI - Lipopolysaccharide decreases scavenger receptor mRNA in vivo. AB - Lipopolysaccharide (LPS) downregulates scavenger receptor (ScR) activity in cultured macrophages through release of tumor necrosis factor-alpha (TNF-alpha). However, LPS administration in vivo stimulates cytokine release from both macrophages and lymphocytes, the combined effects of which could alter ScR expression differently from TNF-alpha in isolation. To investigate whether LPS regulates ScR in vivo, 10 microg/g was injected i.p. into Swiss Webster mice. Administration of LPS produced a profound decrease in hepatic ScR mRNA, with reductions of 74% +/- 8% at 2 h that returned to baseline levels by 6 h. Changes in ScR mRNA abundance coincided with changes in serum concentrations of TNF alpha, which peaked at 2 h (1320 +/- 309 pg/ml) and returned to preinjection concentrations at 4 h. Serum concentrations of interferon-gamma (IFN-gamma) did not increase until 4 h after injection of LPS. There was no effect on ScR mRNA abundance following LPS administration to LPS-resistant strains of mice, C3H/HeJ and IFN-gamma receptor-/-. The LPS-induced reduction in ScR mRNA in Swiss Webster mice was not sufficiently sustained to affect receptor function, as determined by the kinetics of [(125)I]-acetylated LDL clearance from plasma. Therefore, as observed in cultured cells, LPS administration to mice decreases ScR mRNA despite the release of several cytokines in vivo. PMID- 9335437 TI - Aberrantly expressed cytokeratin 1, a tumor-associated autoantigen in papillary thyroid carcinoma. AB - Papillary thyroid carcinoma (PTC) is a somewhat puzzling disease, combining a propensity to metastasize with an indolent clinical course. The often pronounced T cell-dominated inflammatory infiltrate seen in PTC tumors has prompted us to search for signs of a tumor-induced immune response. In previous studies, we have demonstrated large tumor-specific deposits of IgG and complement in PTC tissue and isolated a putative target antigen. This investigation examines the presence of autoantibodies to cytokeratin 1, a high m.w. cytokeratin normally expressed only in suprabasal keratinocytes, in the serum and tumor tissue of PTC patients. Using immunoprecipitation and Western blot, cytokeratin 1-reactive autoantibodies were demonstrated in 80% of the PTC sera tested, and tumor-derived antibodies were shown to precipitate cytokeratin 1. Using immunohistochemistry, cytokeratins 1 and 10 were found in a large proportion of PTC tumors (39/44) but were absent from normal thyrocytes of most PTC-bearing glands. Our results indicate that this protein is expressed aberrantly in neoplastic cells and is immunogenic in this context. PMID- 9335438 TI - Growth patterns of diffuse non-Hodgkin's lymphomas estimated from mitotic and apoptotic indices. AB - Growth rates of neoplasms could be calculated only on the basis of mitotic and apoptotic indices (MI and AI, respectively), assessed on tissue sections, if the duration of mitosis and apoptosis (Tm and Ta, respectively) in vivo were known. For humans, this is practically never the case. What use then can be made of MI and AI to arrive at a relative, crude estimate of the state of growth? As a model system to study this problem, we chose diffusely growing stage I + II non Hodgkin's lymphomas (dNHL, n = 94). Cluster analysis revealed the existence of 3 highly distinct groups of dNHL (clusters I, II and III) in the MI vs. AI per case plot, with a roughly linear relation between both parameters. Most nosologic entities defined by the REAL classification comprise cases that were represented in more than one cluster. We adopted the simple formula GI (growth index) = XMI - AI, where X (= Ta/Tm) remains to be evaluated. Based on the assumption that spontaneous regressions of dNHL are rare but do occur, we estimated that X = 2 or, possibly, 3 are best fits for the pooled dNHLs studied. With the assumption of X = 2, (i) 2MI - AI gave relatively lower values for dNHL than proliferative indices such as %Ki-67+ cells; (ii) values for 2MI/AI per cluster showed a pattern inverse to that for %bcl-2+ cells; and (iii) a plot of 2MI - AI vs. 2MI/AI per case allowed the recognition, especially among NHLs with a low cell turnover, of cases where accumulation of presumably longer-lived cells is an important factor in determining growth. PMID- 9335439 TI - Secondary drug resistance in breast cancer: failure to reverse with oral nifedipine. AB - We tested the efficacy of nifedipine to reverse acquired resistance to chemotherapy regimens containing doxorubicin or vinblastine or both in 12 patients with metastatic breast cancer. All patients had been receiving one or both of these drugs, had had a prior partial response (median duration 5 months, range 2-10) and subsequently progressed. Immediately after drug resistance was documented by tumor progression, eligible patients with measurable or evaluable disease were treated with nifedipine beginning 3 days before restarting the same chemotherapy. The initial dose of nifedipine was 20 mg TID, escalating daily to 40 mg TID on day 3 if the patient had no serious side effects. Nifedipine was continued at the highest tolerable dose during and for 2 days after completion of the chemotherapy. Most patients had < or = 2 prior chemotherapy regimens and a median Zubrod performance status of 1. Twelve patients received a total of 23 courses preceded by nifedipine. No objective tumor responses were observed. The expected toxic effects attributable to nifedipine occurred, but nifedipine did not increase the toxicity caused by the chemotherapy. Nifedipine, given in this dose and schedule, did not reverse acquired drug resistance in patients with breast cancer. PMID- 9335440 TI - Suppression of intracellular Cu-Zn SOD results in enhanced motility and metastasis of Meth A sarcoma cells. AB - We have previously described an inverse relationship between Cu-Zn superoxide dismutase (SOD) activity and invasiveness of a clone of human tongue cancer cells. In these cells, suppression of Cu-Zn SOD activity by transfection with anti-sense cDNA enhanced motility in vitro. The present studies were undertaken to determine whether the inverse relationship between intracellular Cu-Zn SOD activity and motility is a general property of other tumor cells and whether this enzyme indeed defines in vivo metastatic potential. Murine Meth A sarcoma-derived ML-01 cells, which have low metastatic activity, were transfected with anti-sense Cu-Zn SOD cDNA. Two clones with very different SOD activities--ML-AS2, with the most suppressed, and ML-AS5, with the least suppressed activity-were analyzed for their motility and metastatic capability. Compared to the mocktransfectant ML neo, the metastatic potential and motility of the ML-AS2 and ML-AS5 were increased 4.5- and 2.1-fold, respectively. Superoxide treatment enhanced the motility of the AS clones but not that of the ML-neo cells. Our results clearly show that there is an inverse relationship between the intracellular level of Cu Zn SOD, cell motility and in vivo metastatic potential. PMID- 9335441 TI - Coffee and tea intake and risk of cancers of the colon and rectum: a study of 3,530 cases and 7,057 controls. AB - The relationship between coffee, decaffeinated coffee and tea intake and risk of cancers of the colon and rectum was considered combining data from 2 case-control studies, one conducted between 1985 and 1991 in Northern Italy and the other between 1991 and 1996 in 6 Italian centers. Cases were patients below age 80, with histologically confirmed cancer of the colon (n = 2,166) or rectum (n = 1,364), and controls were 7,057 patients admitted to hospital for a wide spectrum of acute, non-neoplastic, non-digestive tract diseases. Compared with coffee non drinkers, the risk of colon cancer was reduced in drinkers of 4 or more cups/day [multivariate odds ratios (ORs) 0.73; 95% confidence intervals 0.60-0.89), with a significant trend in risk with dose; no significant association emerged between coffee drinking and risk of rectal cancer (OR 1.00 for drinkers of 4 or more cups/day). Decaffeinated coffee was consumed in small amounts by about 4% of cases and controls and the OR was 0.92 for colon and 0.88 for rectal cancers. Tea consumption was generally limited to 1 cup/day or to occasional intake and did not substantially modify the risk of colon and rectal cancers. No significant heterogeneity was found for the inverse relationship between coffee intake and colon cancer risk across strata of age at diagnosis, sex, smoking status, total alcohol and meat and vegetable intake, while the protection of coffee was stronger in people eating 3 or more meals/day. Thus, our results confirm that coffee intake has a quantifiable protective effect on colon cancer risk. PMID- 9335442 TI - Melanoma and sun exposure: an overview of published studies. AB - To assess the association between the incidence of cutaneous melanoma; intermittent, occupational and total sun exposure; and history of sunburn at different ages, we conducted a systematic review using results of all published case-control studies which have assessed incident melanoma, sun exposure and sunburn. Twenty-nine studies contributed data on sun exposure and 21 on sunburn. Overall, there was a significant positive association (odds ratio [OR] = 1.71) for intermittent exposure, a significantly reduced risk for heavy occupational exposure (OR = 0.86) and a small, marginally significant excess risk for total exposure (OR = 1.18). There was a significantly increased risk with sunburn at all ages or in adult life (OR = 1.91) and similarly elevated relative risks for sunburn in adolescence (OR = 1.73) and in childhood (OR = 1.95). There was significant heterogeneity with all of these estimates except that of all ages or adult sunburn. These results show the specificity of the positive association between melanoma risk and intermittent sun exposure, in contrast to a reduced risk with high levels of occupational exposure. The association with sunburn also is likely to reflect intermittent exposure; the results do not suggest any strong relationship to age at sunburn. These associations are similar to those reported for basal cell skin cancer but different from those reported for squamous cell cancer. The mechanisms by which intermittent exposure increases risk, while other patterns of exposure do not, remain to be elucidated. PMID- 9335443 TI - Diabetes mellitus and the risk of primary liver cancer. AB - The relationship between diabetes mellitus and primary liver cancer was investigated in a case-control study conducted in Italy between 1984 and 1996 on 428 cases with incident, histologically confirmed hepatocellular carcinoma, 59 with gallbladder and bile duct cancer, and 1,502 control subjects in the hospital for acute non-neoplastic diseases. Sixty-four cases of hepatocellular carcinoma vs. 87 controls reported a history of diabetes, corresponding to an odds ratio (OR) of 2.3 after allowance for age, sex and area of residence, and of 2.1 [95% confidence interval (CI) = 1.4-3.2] after further allowance for alcohol and tobacco consumption, history of hepatitis and liver cirrhosis, body mass index and history of liver cancer in first-degree relatives. The ORs were similar both for subjects diagnosed with diabetes below age 45, who most likely had insulin dependent diabetes, and for those diagnosed later, who were likelier to have non insulin-dependent diabetes. The OR was 2.3 for subjects whose diabetes was diagnosed <5 years before diagnosis of liver cancer, 1.9 for those diagnosed 5-9 years in advance and 2.2 for those diagnosed since 10 years or more. Five cases of gallbladder and bile duct cancer reported a history of diabetes: the corresponding OR was 1.2 (95% CI 0.5-2.9). The OR of hepatocellular carcinoma was 2.4 for males and 2.0 for females, 3.0 for subjects diagnosed with liver cancer under age 60 and 1.8 for those diagnosed at age 60 or over. None of the other covariates considered, including education, history of hepatitis, liver cirrhosis and alcohol drinking showed any meaningful modifying effect or interaction. The potential pathogenic mechanisms include liver alteration-and consequent cell proliferation-in subjects with diabetes. Thus a history of diabetes mellitus could explain about 8% (95% CI 5-11) of cases of liver cancer in this population. PMID- 9335444 TI - Effect of faecal occult blood testing on colorectal mortality: results of a population-based case-control study in the district of Florence, Italy. AB - The aim of our case-control study was to estimate the effect on mortality from colorectal cancer (CRC) of a population-based screening with a faecal occult blood test started in 1982 in a rural area of the district of Florence. We examined the relationship between mortality and the interval since the most recent screening. The cases in the study were 206 individuals who had died from CRC after the age of 41 years. Five controls were selected randomly from the list of individuals alive at the time of diagnosis of the corresponding case and were matched by sex, age and place and length of residence. After adjustment for potentially confounding factors, the odds ratio (OR) for death from CRC for screened persons vs. those not screened was 0.60 [95% confidence interval (CI), 0.4-0.9]. The OR was lowest in the first 3 years after the most recent test (OR = 0.54; 95% CI, 0.3-0.9) and increased towards unity subsequently. Our results suggest that screening for CRC by biennial faecal occult blood testing can reduce mortality from the disease. PMID- 9335445 TI - Local but no systemic immunomodulation by intraperitoneal treatment of advanced ovarian cancer with autologous T lymphocytes re-targeted by a bi-specific monoclonal antibody. AB - We have reported a 27% overall anti-tumor response using i.p. immunotherapy of advanced ovarian carcinoma with autologous, ex vivo expanded, T lymphocytes re targeted with bi-specific monoclonal antibody OC/TR, combined with soluble OC/TR and low-dose recombinant interleukin-2 (IL-2). This treatment had no effect on extraperitoneal disease. Therefore we studied in 13 patients whether this immunotherapeutic protocol resulted only in local or also in systemic immunomodulation. The phenotype of the ex vivo expanded lymphocytes was mainly CD3+, 4-, 8+, 16-, 56-. Their OC/TR-re-targeted cytolytic activity against Igrov 1 ovarian-carcinoma cells was approximately as high in responders as in non responders. Following most therapeutic cycles, the immunophenotype of lymphocytes recovered from the peritoneal fluid was similar to that of the infused T cells (i.e., mainly CD3+, 4-, 8+) and they were coated with OC/TR. However, cytolytic activity of the recovered lymphocytes against Igrov- 1 cells was low in direct assays, and only slightly increased after additional in vitro re-targeting with OC/TR. Systemically, the i.p. immunotherapy resulted in a transient lymphopenia lasting for about 7 days, low (i.e., 5 to 13 ng/ml) serum concentrations of free, functional OC/TR, and very weak coating of circulating T lymphocytes with OC/TR. These peripheral-blood T lymphocytes did not exert OC/TR-re-targeted cytolytic activity. Thus, locoregional OC/TR-re-targeted cellular immunotherapy resulted in substantial local immunomodulation and anti-tumor effects but virtually no systemic immunomodulation. PMID- 9335446 TI - Colorectal cancer mass-screening: estimation of faecal occult blood test sensitivity, taking into account cancer mean sojourn time. AB - Mass screening using the faecal occult blood test (FOBT) can reduce mortality from colorectal cancer. Reliable estimation of FOBT sensitivity is crucial in assessing the potential effectiveness of a mass-screening procedure. Available estimates could be inaccurate because they neglect the temporal aspect of screening. The aim of our study was to estimate the sensitivity of the FOBT in mass screening for colorectal cancer, taking into account the duration of the pre clinical phase of the disease assessed by the mean sojourn time (MST), and to assess whether MST and FOBT sensitivity differ according to cancer subsite. We analysed data taken from the first round of the mass-screening programme of the department of Calvados (France), involving 164,364 subjects of whom 43.4% participated in FOB screening. MST and sensitivity were estimated using a simple empirical approach, a traditional maximum likelihood method and log-linear modelling using the Bayesian technique of Gibbs sampling. MST was estimated as between 4.5 and 5 years for all subsites combined. According to the Gibbs sampling method, MSTs were 3.5, 6.4 and 2.6 years for proximal colon, distal colon and rectal cancer, respectively. Our estimation methods give a low sensitivity for the FOBT (50%), results for different subsites being closer to each other, slightly higher for proximal cancer. Our results strongly suggest that tumour growth rates are very different according to subsite, slowest for distal cancer and speediest for rectal cancer. Consideration of FOBT sensitivity without MST appears unreliable. Our results by subsite suggest that combining FOBT and sigmoidoscopy could be a good strategy for colorectal cancer screening. PMID- 9335447 TI - Detailed microsatellite analysis of chromosome 3p region in non-papillary renal cell carcinomas. AB - Multiple hemi- and homozygous interstitial deletions at chromosome 3p have been found in different types of cancer, including non-papillary renal cell carcinoma (RCC). To determine the frequency and size of deletions, we have analyzed paired normal and tumor DNA obtained from short-term cultures of 104 non-papillary RCCs with 29 microsatellite markers covering the entire chromosome 3p region. Deletion mapping provided evidence for terminal deletion, with the most distal breakpoint between loci D3S1606 and D3S3666 in 94 cases, whereas constitutional heterozygosity was retained at all loci in 4 RCCs. In 6 cases, interstitial deletions were detected. Deletion mapping detected the smallest overlapping region between loci D3S3666 and D3S1560, which corresponds to an approx. 55 cM genetic distance. PMID- 9335448 TI - Foscan-mediated photodynamic therapy for a peritoneal-cancer model: drug distribution and efficacy studies. AB - Distribution of the photosensitizer Foscan (meta-tetrahydroxyphenylchlorin, mTHPC), after i.v. or i.p. injection, was investigated in Wag/Rij rats bearing i.p. tumours. These results were compared with the efficacy of mTHPC-mediated photodynamic therapy for illumination intervals of 4 hr to 3 days. For the distribution experiments a single tumour (CC53I colon carcinoma) was implanted intra-abdominally in a fat pad, or a cell suspension (1 x 10(6) CC531 cells) was injected into the peritoneal cavity, which results in a dissemination of tumour nodules on the peritoneum. 14C-mTHPC was not selectively taken up in the single tumour model after i.v. or i.p. injection, but higher concentrations were achieved for i.p. administration. For this tumour model the concentration ratios between tumour and normal tissue never exceeded a value of 3. In the disseminated tumour model, an uptake of up to 40% of the injected dose was found per gram tumour at 4 hr after an i.p. injection and this resulted in very high (> 14) concentration ratios of tumour to normal tissues. Low uptake was found after the i.v. injection route (1% of the injected dose per gram tumour) with lower tumour/normal tissue ratios (<8). The efficacy of i.p. photodynamic therapy (IPPDT) was evaluated using the single-tumour model only. The lower abdomen was illuminated at 4 hr to 3 days after mTHPC, and tumour size was repeatedly measured via a small laparoscopy. Significant delay in tumour regrowth was achieved for 6 J x cm-2 at 1 day after i.v., or at 4 hr after i.p. mTHPC (p values 0.019 and 0.045 respectively). Response to PDT, of tumours implanted in the fat pad, was not greater for i.p. administration of the photosensitizer and there was a poor correlation between times of maximum drug uptake in tumours and optimal illumination times for PDT efficacy. PMID- 9335450 TI - Effects of peroxisome proliferators and 12-O-tetradecanoyl phorbol-13-acetate on intercellular communication and connexin43 in two hamster fibroblast systems. AB - Effects of 12-O-tetradecanoyl phorbol 13-acetate (TPA) and the hepatic peroxisome proliferators (HPPs) clofibrate, methyl clofenapate (MCP), di(2 ethylhexyl)phthalate (DEHP) and mono(2-ethylhexyl)phthalate (MEHP) were studied in 2 gap junctional intercellular communication (GJIC) systems, metabolic cooperation in V79 cells and microinjection/dye transfer in Syrian hamster embryo (SHE) cells and V79 cells. TPA inhibited GJIC in both systems but was considerably more potent in V79 cells. SHE cells showed a rapid and transient inhibition of GJIC after exposure to HPPs, with maximal inhibition occurring at 5 15 min. The transient inhibition could be caused by metabolization of the compounds. Clofibrate and MEHP produced strong inhibition of metabolic cooperation in V79 cells at high concentrations, while the effect of MCP and DEHP was lower. However, DEHP, MEHP and clofibrate strongly inhibited dye transfer in V79 cells after a 30 min exposure. Clofibrate also showed a dose- and time dependent effect on dye transfer in V79 cells. The phosphorylation status of the gap junction protein connexin43 (Cx43) changed minimally in SHE cells after exposure to TPA or HPPs. Cx43 from V79 cells was strongly affected by TPA, but not by HPPs. Immunofluorescence of Cx43 disappeared in both cell types when they were exposed to TPA and MEHP, but not to the other HPPs. Thus, there is no direct correlation between the inhibition of GJIC and changes in the phosphorylation status of Cx43 or the appearance of Cx43 in immunofluorescence experiments. The discrepancies may partly be explained by binding of accessory proteins to Cx43. We point out sequences that may be involved in such binding. PMID- 9335449 TI - Does tumour uptake of Foscan determine PDT efficacy? AB - Preferential retention of photosensitizers in tumours has always been one of the major goals in the search for new photosensitizers and has determined the design of clinical trials with respect to the interval between drug administration and illumination. The purpose of this study was to investigate the importance of tumour and plasma concentrations of Foscan (mTHPC, meta tetrahydroxyphenylchlorin) in relation to PDT effect. Both pharmacokinetic and tumour response studies were carried out in mice bearing s.c. RIF1 tumours. mTHPC was injected in 1 or 2 doses of 0.3 mg x kg-1. For distribution studies, 14C labelled mTHPC was given 5 min to 48 hr before determination of plasma and tumour drug levels. Non-labelled sensitizer was used to determine the PDT efficacy for illumination at 5 min to 48 hr after drug administration. PDT efficacy was greatest for illumination at 1 to 3 hr, and for an interval of 48 hr there was no significant tumour-growth delay. In contrast, mTHPC tumour drug levels reached a maximum 6 hr after injection and remained high for 48 hr. A comparison of pharmacokinetics and response studies revealed no significant correlation between tumour mTHPC levels and tumour response. There was, however, a significant correlation between plasma drug levels and tumour response for time intervals of 1 to 48 hr. This association may imply that PDT protocols should use shorter drug light intervals in combination with lower drug doses. This would increase safety and decrease the extent and duration of normal tissue photosensitization. PMID- 9335452 TI - Adenovirus-mediated gene transfer of fibroblast growth factor-1: angiogenesis and tumorigenicity in nude mice. AB - Gene transfer of angiogenic growth factors with replication-deficient recombinant adenovirus (Ad) vectors may provide a new approach to the treatment of ischemic diseases. To determine if Ad-infected cells could stimulate angiogenesis in vivo and to assess the tumorigenicity of cells infected with these vectors, NIH3T3 fibroblasts infected with Ad vectors coding for human acidic fibroblast growth factor (aFGF-1) were used in angiogenic and tumorigenic assays. Infected cells induced a strong angiogenic response in vivo, while cells infected with control virus did not. Stable 3T3 transfectants expressing the FGF-1 gene were also highly angiogenic and exhibited growth in soft agar, while Ad-infected cells did not. Ad-infected cells grew transiently in nude mice, whereas 3T3 transfectants formed large tumors which grew exponentially. Extrapolation of cell dose-response curves showed that a minimum of 1.5 x 10(4) infected cells were required for transient tumor cell growth in vivo. Ad-infected cells cultured in vitro for 30 days lost their invasive phenotype and the ability for transient cell growth in nude mice. Thus, phenotypic changes induced by Ad-mediated gene transfer of FGF-1 are transient both in vitro and in vivo, suggesting that these Ad vectors do not have tumorigenic potential. Stimulation of angiogenesis by Ad-infected cells may be useful for the evaluation of anti-angiogenic and anti-tumor agents. PMID- 9335451 TI - Identification of novel drug resistance-associated proteins by a panel of rat monoclonal antibodies. AB - Since some multidrug-resistant (MDR) tumor cell lines show drug accumulation defects but do not over-express Pgp or MDR protein (MRP), a search was made for novel MDR-related transporter proteins by immunizing rats with non-small cell lung cancer SW- 1573/2R120 cells to produce monoclonal antibodies (MAbs). Five rat MAbs (LMR-4, -12, -42, -44 and -94) were generated, showing strong membranous staining of non-Pgp MDR SW- 1573/2R120 tumor cells and minimal reactivity to the corresponding parental and revertant cell lines. In addition, a 6th MAb (LMR-5) was isolated, recognizing the MDR-related lung resistance protein (LRP), previously identified as the major vault protein. The first 5 LMR MAbs show predominantly membranous staining of several non-Pgp MDR tumor cell lines of different histogenetic origins, except for LMR-4, which recognizes only MDR sublines of the SW- 1573 cell line. Flow-cytometric analysis revealed that all MAbs, except LMR-4 and -5, detect outside epitopes. Functional studies showed that these MAbs did not restore the daunorubicin accumulation defect. All but one of the MAbs (LMR-42) showed staining of distinct normal human tissues, notably epithelial cells lining the airways and digestive tract. In addition, staining of vascular endothelial cells was found with all MAbs except LMR-4. Three MAbs (LMR 12, -44 and -94) showed remarkable immunoreactivity with vincristine-selected SW- 1573 sublines. By immunoblotting and precipitation, the LMR antigens were found to be in the 42-69 kDa range. PMID- 9335453 TI - Activation status and function of the VLA-4 (alpha4beta1) integrin expressed on human melanoma cell lines. AB - We have examined the functional status of the VLA-4/alpha4beta1 integrin in a panel of human melanoma cell lines, focusing on the ability of cells expressing alpha4beta1 to mediate adhesion to the alpha4-specific ligands CS-1 peptide and VCAM-1. All melanoma cells expressing alpha4pbeta1 (8 of 10 lines examined) were capable of adhering to these specific ligands in adhesion assays, whereas 2 cell lines (HMB2 and VUP) which lacked surface alpha4 were unable to do so. Adherence of different melanoma cell lines to VCAM-1 was relatively uniform and not susceptible to upregulation with known integrin-activating factors, such as manganese ions, phorbol ester and activating monoclonal antibody (mAb) TS2/16. Cell adhesion to CS-1 peptide, however, varied according to cell surface receptor density and, in some cases, could be up-regulated by integrin-activating factors. Adhesion of SK23 cells to CS-1 peptide was increased by all 3 activating stimuli, whereas for all other melanoma cells an increase was obtained only by the use of TS2/16 mAb. Our data indicate not only an unusually low activation state of alpha4beta1 in SK23 cells but also heterogeneity in the activating capacity of the various stimuli. Moreover, a protein kinase C-dependent role in alpha4beta1 activity was suggested by adhesion assays carried out in the presence of the protein kinase C inhibitor calphostin C, which considerably reduced adhesion to CS-1 peptide. PMID- 9335454 TI - Relationship between exposure to TPA and appearance of transformed cells in MNNG initiated transformation of BALB/c 3T3 cells. AB - In the BALB/c-3T3-cell transformation system, the effect of 12-O tetradecanoylphorbol-13-acetate (TPA) exposure on the appearance of transformed cells was examined in order to investigate the mechanisms of in vitro tumor promotion. Optimal duration of TPA exposure on N-methyl-N'-nitro-N nitrosoguanidine(MNNG)-initiated cells was at least 11 days. To investigate the effect of transformation frequencies of altering inoculating cell density at the replating of MNNG-exposed cells and of altering the time of starting TPA exposure, MNNG-exposed cells were replated at various inoculum sizes. With lower inoculum sizes (1 x 10(3) to 3 x 10(4) cells/dish), maximum TPA-induced transformation occurred for TPA commencement at confluence, while with higher inoculum size (1 x 10(5) cells/dish), maximum transformation frequency was observed when TPA exposure was started on day 7 after replating, being some 2 days after confluence. This may suggest that there are different mechanisms involved, depending on inoculum size, and that these may involve cell-cell interactions (at lower inoculum) and mutation expression periods (at higher inoculum). By means of redispersion experiments, it was demonstrated that the appearance of transformed cells begins on about day 7 after replating at a cell density of 1 x 10(4) cells/dish. These results suggest the usefulness of the replating method for optimizing transformation in the BALB/c-3T3-cell transformation assay, and provide insight into the time frame of expression of MNNG-initiated transformants and TPA-induced expansion of these transformants. PMID- 9335455 TI - Anti-sense oligonucleotides directed against EGF-related growth factors enhance anti-proliferative effect of conventional anti-tumor drugs in human colon-cancer cells. AB - We have demonstrated that anti-sense phosphorothioate oligodeoxynucleotides (AS S oligos) directed against the EGF-like growth factors CRIPTO (CR), amphiregulin (AR) or transforming-growth-factor-alpha(TGFalpha) mRNA, are equipotent in their ability to inhibit the growth of human colon-carcinoma GEO cells. In this study, we evaluated the effect of combinations of these AS S-oligos and conventional anti tumor drugs, such as 5-fluorouracil (5-FU), adriamycin (ADR), mitomycin C (MIT) and cis-platinum (CDDP), on GEO cell growth. Dose-dependent growth inhibition was observed by treatment either with AS S-oligos or with anti-tumor drugs, using a clonogenic assay. Furthermore, an additive growth inhibitory effect occurred when GEO cells were exposed to the AS S-oligos after treatment with different concentrations of either 5-FU, MIT, ADR or CDDP. For example, treatment of GEO cells with a combination of low concentrations of 5-FU and any of the 3 AS S-oligos resulted in up to 70% growth inhibition. However, treatment of GEO cells with AS S-oligos before exposure to 5-FU or CDDP resulted in reduced efficacy of both drugs. Flow-cytometric analysis of DNA content demonstrated that treatment with the AS S-oligos caused a slight reduction of the percentage of cells in the S-phase of the cell cycle. These data suggest that combinations of AS S-oligos directed against EGF-related growth factors and of conventional anti tumor drugs may result in efficient inhibition of colon-carcinoma cell growth. PMID- 9335456 TI - Vascular response of tumour and normal tissues to endothelin-1 following antagonism of ET(A) and ET(B) receptors in anaesthetised rats. AB - Modification of blood flow by endothelin-1 (ET-1) was examined in the s.c. HSN fibrosarcoma and compared to normal tissues of anaesthetised CBH/CBi rats. The ET receptor subtypes involved in the response were investigated using the ET(A) and ET(B) receptor antagonists BQ-610 and BQ-788, respectively. Blood flow and vascular resistance were determined using the uptake of radiolabelled iodo antipyrine (125I-IAP). BQ-610 or BQ-788 was infused for 30 min prior to blood flow determination. ET-1 was administered 15 min into the infusion time. BQ-610 and BQ-788 infused alone did not modify any vascular parameters. Tumour blood flow increased slightly following ET-1, contrasting with most normal tissues, in which blood flow was reduced. Vascular resistance increased in all tissues, including the tumour. Neither antagonist significantly modified the ET-1-induced changes in tumour blood flow or vascular resistance, whereas in the majority of normal tissues BQ-610 attenuated and BQ-788 potentiated the vascular resonse to ET-1. Our results show that the HSN tumour vasculature is only weakly responsive to ET- 1 and antagonism of its effects by BQ-610 and BQ-788. This contrasts with the majority of normal tissues, in which ET- 1 induces an intense vasoconstriction. PMID- 9335457 TI - Modulation of human stromelysin 3 promoter activity and gene expression by human breast cancer cells. AB - The matrix-degrading enzyme family of matrix-metalloproteinases (MMPs) has been implicated in the process of tumour metastasis. Cellular protein and RNA localisation techniques have been used to show that, whilst several MMP genes are expressed in both cancer and stromal cells, stromelysin 3 is expressed only in stromal fibroblasts adjacent to cancer cells. Immunohistochemical and in situ hybridisation evidence suggests that neoplastic cells can stimulate stromal cell MMP production either in a paracrine fashion or by a cell-cell contact mechanism. Using 2 different lengths of the human stromelysin 3 (ST3) gene 5' flanking sequence cloned upstream of luciferase and CAT reporter genes, we now show that human breast cancer cells can directly activate the ST3 promoter. The putative response element in the ST3 promoter, which lies between 0.46 and 3.4 kb upstream of the transcription start site, is able to effect a 2- to 3-fold increase in downstream gene expression. We further show that this transcriptional up regulation definitely occurs via a paracrine, and possibly via a cell-cell contact, mechanism. Confirmation that this ST3 promoter activation results in ST3 gene induction of a similar magnitude was shown using Northern blotting of stimulated fibroblasts. Our data provide further evidence that cancer cells can induce fibroblast MMP expression and help to explain the in vivo expression pattern of ST3 in breast cancer. PMID- 9335458 TI - Down-regulation of the nm23.h1 gene inhibits cell proliferation. AB - nm23 gene expression is strictly related to the state of cell growth. The level of its expression parallels the fraction of thymidine-incorporating cells (S phase cells) in neoplastic mammary tissues and in the synchronously cycling fraction of MCF 1OA cells. nm23.h1 reaches a peak of expression in the S-phase, and is present at very low level during the GO/G1 phase. Two strategies are used to demonstrate the direct involvement of the nm23.h1 gene in the process of cell proliferation. The first consists of transient inhibition of nm23.h1 expression by using anti-sense oligonucleotide treatment; weak inhibitory effect on cell proliferation is observed. The second strategy involves the stable inhibition of nm23.h1 expression by transfection of MCF1OA cells with a plasmid vector expressing the human nm23.h1 anti-sense mRNA. The anti-sense-transfected cells show consistently slower proliferative activity than the control. PMID- 9335459 TI - Human herpesvirus-8 (HHV-8) DNA associated with anaplastic large cell lymphoma of the B-cell type in an HIV-1-positive patient. PMID- 9335460 TI - Immunohistochemical and ultrastructural investigation of the human vestibular dark cell area: roles of subepithelial capillaries and T lymphocyte-melanophage interaction in an immune surveillance system. AB - BACKGROUND: The aim of the present study was to morphologically characterize the structure of the subepithelial blood vessels in the dark cell area of the human vestibular organs, and to determine whether immunocompetent cells such as macrophages and lymphocytes could be found around these small blood vessels. MATERIALS AND METHODS: All 31 surgical specimens (semicircular canals and utricles) were obtained from patients with vestibular schwannoma. Formalin fixed specimens were stained with hematoxylin and eosin (H&E), and with antibodies to von Willebrand Factor (vWF), leukocyte common antigen (LCA), and UCHL-1, and were examined with light microscope. Specimens fixed with glutaraldehyde were examined with a transmission electron microscope (TEM). OBJECTIVES: Subepithelial blood vessels stained positive for vWF. By TEM observation, these blood vessels were observed to be capillaries that consisted of non-fenestrated endothelium, occasional pericytes, and a basement membrane. They were usually accompanied by melanophages with a number of secondary lysosomes containing phagocytosed degraded melanosomes and lipid droplets. Moreover, melanocytes and their cell processes directly surrounded these subepithelial capillaries. The fact that cells which were positively stained with LCA and UCHL-1 were present both in the intra- and subepithelial layer of the specimens, and that by TEM the intra- and subepithelial mononuclear cells with a lymphoid appearance had clustered dense bodies in their cytoplasm, suggested that they were a population of T lymphocytes. CONCLUSIONS: Results suggested the possibility of a T lymphocyte melanophage (macrophage) interaction, both originating from and harbored around subepithelial capillaries, which suggests the presence of an immune surveillance system in the human vestibular organs. PMID- 9335461 TI - Histologic evidence: growth hormone completely prevents reduction in cortical bone gain and partially prevents cancellous osteopenia in the tibia of hypophysectomized rats. AB - BACKGROUND: In previous studies we found that the cause of bone loss in young hypophysectomized (HX) animals was due primarily to an inhibition in growth dependent bone gain and a decrease in bone turnover. The aim of this study was to determine whether growth hormone, which has stimulatory effects on bone growth and turnover, can prevent HX-induced skeletal alterations in rats. METHODS: Female Sprague-Dawley rats were divided into baseline control (BASAL), age matched control (CON), HX, HX plus low-dose GH (1.5 mg/kg/d, subcutaneously), and HX plus high-dose GH (4.5 mg/kg/d) groups. The BASAL group was sacrificed at 2 months of age and the remaining groups were sacrificed after 6 weeks of treatment. Cancellous and cortical bone histomorphometry was performed on double fluorescent-labeled 40 microm-thick sections of the proximal tibia and tibial shaft. RESULTS: Both low- and high-dose GH prevented the HX-induced decrease of IGF-I serum levels. High-dose GH also significantly increased the body weight and the wet weight of the gastrocnemius muscle when compared to the CON groups. In the tibial shaft, the periosteal labeled surface, mineral apposition rate and bone formation rate were higher in both of the GH-treated groups than in the HX group (P < 0.05). The tissue area and cortical bone area of the high-dose GH treated rats were greater than those of the HX rats, but did not differ from those of the CON rats. In the proximal tibia, both low- and high-dose GH prevented an HX-induced decrease in the longitudinal growth rate and growth plate width, and increased surface-based bone formation compared to the HX and CON. Cancellous bone volume, tissue-based bone formation rate, and eroded surface in both of the GH-treated groups were higher than those of the HX group, but lower than those of the BASAL and CON groups (P < 0.05). Bone architecture of the HX rats was also improved after GH treatment. CONCLUSIONS: This study clearly demonstrates that GH replacement at the dosage of 4.5 mg/kg/d can completely prevent the HX-induced reduction in cortical bone gain in the tibial shaft, but can only partially prevent cancellous osteopenia in the proximal tibia after six weeks. PMID- 9335462 TI - Aging- and ovariectomy-related skeletal changes in spontaneously hypertensive rats. AB - BACKGROUND: The skeletal impact of estrogen deficiency on subjects with hypertension has not been studied previously. In this study, we examined the skeletal characteristics of female spontaneously hypertensive rats (SHR) and their normotensive genetic control Wistar-Kyoto rat (WKY). We aimed to reveal: 1) the skeletal characters of female SHR, and 2) the response of SHR to ovariectomy (ovx) when compared to WKY and other strains. METHODS: Undecalcified double fluorescent labeled cancellous (proximal tibial metaphysis, PTM) and cortical (tibial shaft, TX) bones from 23-weeks-old, and from rats 2 and 8 weeks post-ovx were studied. RESULTS: The SHR showed lower body weight, higher heart rate, and higher blood pressure than the WKY. Female SHR possessed more percent cancellous bone, less net cortical bone, smaller tissue area, and thinner cortex than WKY. Furthermore, SHR exhibited an age-related cancellous (-18%) and cortical (-7%) bone loss associated with a decrease in the longitudinal growth rate and bone balance and a decrease in periosteal bone formation in cortical bone. In contrast, the WKY maintained most of these parameters unchanged at their 23-week old levels. Ovariectomy induced earlier and greater cancellous bone loss in the SHR than in the WKY, with greater increases in bone turnover rate, eroded surface, activation frequency, and a decrease in the ratio of labeled to eroded perimeter in PTM at 2 weeks postsurgery. However, the two groups exhibited no differences in bone loss at 8 weeks after ovx in PTM and TX. CONCLUSIONS: Spontaneously hypertensive rats were highly sensitive to estrogen deficiency. This might have clinical relevance to those postmenopausal women who suffer from hypertension, in that they may be more susceptible to osteopenia. If so, preventive measures should be initiated sooner than otherwise. PMID- 9335463 TI - Effects of maternal bilateral ureteral ligation on formation of the glomerular basement membrane in fetal rat kidney. AB - BACKGROUND: The present study was designed to clarify development of filtration property of fetal renal glomerulus when maternal kidney is dysfunctional. MATERIALS AND METHODS: Maternal bilateral ureteral ligation was performed on days 17, 19, and 21 of pregnancy. One day after each operation, cationized ferritin (CF), native ferritin (NF), and horseradish peroxidase (HRP) were injected, respectively, in the fetuses. Distribution of the tracers in the fetal kidneys was investigated electron microscopically. RESULTS: On fetal day 20, clustered CF particles were present in the laminae rarae interna and externa of the glomerular basement membrane of the fetuses from ureter-ligated mothers, while the clusters were arrayed in three to four layers in that of the fetuses from sham-ligated ones. On fetal day 22, a relatively large amount of CF particles was present in the lamina rara externa in the fetuses from ureter-ligated mothers when compared to that in the age-matched control fetuses. On fetal days 20 and 22, the number of NF particles was decreased, and shortening of the time for filtration of HRP through the glomerular basement membrane was observed in the fetuses from the ligated mothers. CONCLUSIONS: These results suggest that dysfunction of maternal kidneys causes accelerated formation of fetal glomerular basement membrane and stimulates glomerular function in filtration in fetal rat kidney. PMID- 9335464 TI - Insulin-like growth factor I receptor gene expression during postnatal development of rabbit kidney. AB - BACKGROUND: Insulin-like growth factor-I (IGF-I) is a peptide growth factor whose biological effects are mediated through a specific receptor (IGF-IR). IGF-I and IGF-IR are detected in both fetal and adult kidneys and have both metabolic and growth effects. IGF-IR expression during postnatal kidney development is not well defined and the biological role of this receptor during the postnatal stage is not clearly established. The purpose of the present study was to analyze IGF-IR gene expression during the postnatal development of rabbit kidney to achieve a better understanding of the correlation between growth and differentiation of kidney tissues and IGF-IR expression. METHODS: Using in situ hybridization, we studied changes in IGF-IR expression in the kidneys of newborn rabbits and those up to 35 days old. Evaluation of the stage of kidney development and morphological maturation was made on histological sections stained with hematoxylin-eosin. RESULTS: High levels of IGF-IR gene expression in the rabbit kidney occurred in the last stages of postnatal development and in the adult stages; during the development of the subcapsular metanephrogenic zone, IGF-IR gene expression was not observed. IGF-IR mRNA was expressed by proximal and distal tubules and by collecting ducts after these tissues attained morphological maturation. The appearance of IGF-IR mRNA in these kidney structures followed a precise temporo-spatial sequence. IGF-IR was not expressed by renal corpuscles, Henle's loops, inner medullary collecting ducts, vessels, or interstitial cells at any study stage. CONCLUSIONS: The temporal and spatial patterns of IGF-IR gene expression during postnatal development of the rabbit kidney suggest that IGF-IR and its ligands are relevant for the acquisition of the function, and not for development events, by proximal and distal tubules and collecting ducts. This study also suggests that IGF-IR mRNA localization constitutes a useful marker to determine the functional maturation of these renal structures. PMID- 9335465 TI - Ultrastructure of the salivary glands in the midtongue of the common vampire bat, Desmodus rotundus. AB - BACKGROUND: All examined mammals have at least two sets of lingual salivary glands: von Ebner's glands and Weber's glands. A third set, the glands of Blandin and Nuhn, is present in the tongues of some but not all mammals. Vampire bats, Desmodus rotundus, are unusual in that they possess another set of lingual glands, these being in the midtongue region. METHODS: The anterior half of the tongue was extirpated from several adult vampire bats, dissected, and tissue blocks derived from the midregions of the body of the tongue prepared for transmission electron microscopy by conventional means. RESULTS: The midlingual glands are in the form of long, tubular secretory endpieces that are succeeded by ducts of simple morphology. In general, the secretory portions consist of two cell types, which may be intermingled in the same tubule or may form tubules that consist wholly of one cell type or the other. Seromucous cells usually have one or several rough endoplasmic reticulum cisternae that are hugely distended by a homogeneously dense material. Their granules have a bizonal substructure: one or several dense bands are embedded in a lighter matrix. Mucous cells are rather typical in structure, but their secretory product is different from run-of-the mill mucous droplets. These droplets vary in density from cell to cell. In some cells, these droplets have a relatively light matrix; in other cells, the droplet is unusually dense, consisting mainly of a dark, structureless matrix with marginal lenticular lacunae of low density in which some short, irregular filaments are scattered. A rare finding is the presence of ciliated cells intermingled with secretory endpiece cells. The cilia are of conventional morphology. Secretory tubules are succeeded by ducts that resemble intercalated ducts; the epithelium of these ducts gradually increases in height to form a kind of excretory duct, without the intervention of striated ducts. As the ducts approach the lingual surface, the epithelium changes to stratified squamous. CONCLUSIONS: Saliva produced by the midlingual glands may be an aid in the reciprocal grooming behavior of vampire bats. Based on their morphology, the excurrent ducts may not modify the initial saliva elaborated by these glands and might act simply as pipelines by which the saliva reaches the mouth. PMID- 9335466 TI - Incidence of air pollution in the pulmonary surfactant system of the pigeon (Columba livia). AB - BACKGROUND: The integrity of both pulmonary surfactant and surfactant producing cells, type II pneumocytes, is essential for normal pulmonary function. Almost all studies about air pollution effects on the pulmonary surfactant system have been performed by biochemical techniques, using atmospheric pollutants in a much higher concentration than that found in polluted city air. A comparative study of the number of lamellar bodies (LB) (pulmonary surfactant precursors) from type II pneumocytes of pigeon lungs belonging to a contaminated city habitat and a noncontaminated countryside one has been carried out. METHODS: Lungs of both pigeon groups were subjected to standard processing for light microscopy. A representative number of histological sections from each lung were stained by Gomori's methenamine-silver nitrate technique, which appears to be a good marker for pulmonary LB. Diverse areas of atrial and parabronchial epithelia from each section were photographed, and then quantification of the number of LB per type II pneumocyte section was performed. The data were statistically analyzed and compared on the basis of the levels of atmospheric pollutants. RESULTS: Gomori's methenamine-silver nitrate technique is a useful staining method to study pneumocyte LB. The average of LB per cell section in city birds was about 33% less than in countryside specimens. Moreover, a great amount of macrophages in the lungs from city pigeons has been shown to be present, whereas this cell type is absent or very scarce in the lungs from country birds. CONCLUSIONS: Solid particles and high rates of nitrogen oxides in the polluted air, as well as the oxygen metabolites produced by the macrophages, would represent the main factors for the marked decrease in the amount of LB from type II pneumocytes. PMID- 9335467 TI - An electron microscopic study of the parabronchial epithelium in the mature lung of four bird species. AB - BACKGROUND: No integrated comprehensive description of the ultrastructure of the parabronchial epithelium is available. The origin, discharge, and occurrence of the trilaminar substance have not yet been sufficiently studied. Therefore, the main objectives were to classify the cell types of the parabronchial epithelium and to describe their role in manufacturing the trilaminar substance. METHODS: Lung tissue of mature quail, domestic fowl, town pigeon, homing pigeon, and barn owl was subjected to standard processing for transmission electron microscopy, both after intratracheal inflation and intravascular perfusion. RESULTS: The atrial epithelium is constituted by granular and squamous atrial cells. Granular cells (1) are confined to the atrial wall; they produce and discharge osmiophilic lamellar bodies. Squamous atrial cells (2) manufacture and discharge a trilaminar substance in sheets sandwiched between the long microvilli emerging from the apical cytoplasm. Their attenuating cell outgrowths overlap granular cells. At the bases of atria, they pass as intermediate squamous atrial cells to the infundibula, contacting the extensions of squamous respiratory cells. The squamous atrial cells undergo distinct structural variations depending on age and environment. Squamous respiratory cells (3) (cellulae squamosae) continuously line the air capillaries and neighboring infundibula. They constitute the epithelial compartment of the blood-gas barrier. The cell bodies extend long, very thin cell outgrowths. The apical surface is smooth and the basal part is invested with a very thin basement membrane. The trilaminar substance originates from granular and agranular endoplasmic reticulum in the form of convoluted profiles which are discharged as an acellular lining layer on the air surface of squamous respiratory cells. CONCLUSIONS: Granular cells are analogous to the type II cells of mammalian pulmonary alveolus. Squamous atrial and respiratory cells, of a common embryonic origin, do not meet any counterpart in epithelial cell populations of lung terminal airways in vertebrates. The specific trilaminar substance--lipoproteinaceous in nature--is a constant compound of atria and air capillaries. PMID- 9335468 TI - Neurotrophin receptor-like protein immunoreactivity in human lymph nodes. AB - BACKGROUND: Trk proteins are essential constituents of the high-affinity signal transducing neurotrophin receptors. They are expressed in a variety of non neuronal tissues, including lymphoid organs, but their cellular localization in these remains to be established, as does the exact role of neurotrophins in the immune system. In this study we used immunohistochemical methods to analyze the cellular distribution of TrkA, TrkB, TrkC, and p75 (the low-affinity pan neurotrophin receptor) proteins in normal human lymph nodes. METHODS: Formaldehyde-fixed, paraffin-embedded human lymph nodes were processed for indirect immunoperoxidase labelling, using antibodies against each Trk protein, human p75, and a panel of antibodies against B-lymphocytes (CD20), macrophages (MAC387), dendritic cells (S-100 protein). RESULTS: Immunoreactivity (IR) for p75 was observed in follicular dendritic cells of lymphoid follicles, and possibly in B cells. TrkA-like IR was seen in dendritic cells and also in some follicular dendritic cells, and in blood vessel walls. TrKB-like IR labelled scattered cells, mostly in the T cell zones, identified as macrophages, while specific TrkC like IR could not be observed in immunocompetent cells. In no case was Trk-like IR seen in lymphocytes. CONCLUSIONS: These results demonstrate the occurrence of Trk-like proteins in normal human lymph nodes and describe their cellular localization, favoring the notion that neurotrophins have a physiological role in the immune system, possibly acting through accessory cells and not directly on lymphocytes. PMID- 9335469 TI - Development of mandibular cartilages in the rat. AB - BACKGROUND: The mammalian mandible develops around Meckel's cartilage and other secondary cartilages, including the dentary. There have already been many studies of the development of the rat mandible that have employed histological serial sections. However, no previous investigators have captured the three-dimensional features of the developmental process. METHODS: In this study, the technique of double staining with alizarin red S and alcian blue was employed directly on whole body specimens to investigate the three-dimensional development of the rat mandible. RESULTS AND CONCLUSIONS: We found that the molar socket obstructs the developing mandible, causing it to emerge medial to Meckel's cartilage. Our results also indicate that the secondary mandibular cartilages may contribute to supplementary growth in response to local factors. PMID- 9335470 TI - Timing and embryology of esophageal atresia and tracheo-esophageal fistula. AB - BACKGROUND: The embryology of tracheo-esophageal anomalies is controversial. The development of an adriamycin-treated animal model has enabled improved understanding of the embryogenesis of these anomalies. Using this model, we aimed to describe the events leading to esophageal atresia and tracheo-esophageal fistula. METHODS: Timed-pregnant Sprague-Dawley rats were injected daily with adriamycin intraperitoneally at a dose of 2 mg/Kg on days 6-9 of gestation. Histological sections were prepared from 96 experimental and 34 control rat embryos at 11-14 days gestation (plug day = day 0). RESULTS: The tracheal bud failed to develop normally from the foregut, leaving the foregut to give origin to both bronchi and differentiate into the respiratory system, and then continue as a fistula to the lower esophageal segment. Dorsal pouching of the proximal foregut, which is seen clearly on day 13, is responsible for the development of the upper esophageal segment. CONCLUSIONS: We conclude that failure of the tracheal bud to develop normally from the primitive foregut is the main event which leads to the tracheo-esophageal anomalies. As the proximal part of the primitive foregut develops primarily into a trachea rather than an esophagus, the anomaly of the esophagus could be described as agenesis instead of atresia. PMID- 9335471 TI - Development of Meckel's cartilage in the symphyseal region in man. AB - BACKGROUND: The aim of this work is to clarify the aspects which are at present most controversial about the development of the anterior segments of Meckel's cartilage, such as the role of and determination of the area that is incorporated in the development of the human mandible. METHODS: Light microscope studies were done on 25 embryos and human fetuses from the collection of the Institute of Embryology at the University Complutense of Madrid and the Department of Morphological Science from the University of Granada. Specimen length was between 18 and 125 mm crown-rump. RESULTS: During the embryonic period, Meckel's cartilages were placed in the midline of the mandibular arch but fusion was not observed between them. Ossification of Meckel's cartilage begins at the end of the embryonic period and is completed in the fetal period and the portion that participates in mandibular formation is determined. This segment extends from the mental foramen to near the midline of the mandible. In this region, on the dorsal surface of the symphysis, cartilaginous nodules that originate from Meckel's cartilage are isolated. CONCLUSIONS: The ventral portions of Meckel's cartilage do not fuse in the midline of the mandibular arch. These present endo- and perichondral ossification and the section from the mental foramen to near the midline (mandibular symphysis) participates in mandibular formation. The ventral ends of Meckel's cartilage, i.e., the ends nearest the midline, do not ossify and remain isolated on the dorsal surface of the fetal mandibular symphysis. PMID- 9335472 TI - Mesencephalic trigeminal nucleus neurons supplying the jaw closing muscles have no spinal projection: a fluorescent double-labeling study in birds and mammals. AB - BACKGROUND: The present study deals with the possibility that the mesencephalic trigeminal nucleus (MeV) neurons that innervate the muscle spindles of the jaw closing muscles may also have collaterals projecting to the cervical spinal cord. At the same time, we reexamine the morphology of these cells and their location within the MeV. METHODS: The fluorescent retrograde tracers Fast Blue (FB) and Diamidino Yellow dihydrochloride (DY) were injected into the jaw closing muscles and C2-C3 spinal cord segments, respectively, of duck, rat, and rabbit in one series of experiments. In a second series of animals, the targets of the tracers were reversed. RESULTS: Retrogradely double-labeled cells (FB+DY) were not found in the MeV. On the contrary, the tracer injected into the muscles retrogradely labeled only large unipolar MeV cells, whereas the tracer injected into C2-C3 spinal cord segments labeled only small multipolar cells which were intermingled with the MeV somata of muscle spindle afferents. CONCLUSIONS: These findings exclude the possibility of spinal projections via collaterals of MeV cells supplying muscle spindles of jaw closing muscles in duck, rat, and rabbit. Moreover, the retrograde double-labeling technique evidences two cellular populations within the MeV of the duck, rat, and rabbit: large unipolar neurons which are the cell bodies of primary afferents from jaw closing muscles and small multipolar cells projecting to the upper cervical spinal cord. PMID- 9335473 TI - Organization of the zona incerta in the macaque: an electron microscopic study. AB - BACKGROUND: The zona incerta (ZI) receives projections from many telencephalic and brainstem structures. On the basis of its connectivity and physiology, this nucleus has been implicated in the control of saccadic eye movements. Because of the complexity of its afferent signals and its simple efferent signal, there must be much local interaction within the ZI to integrate these various afferents. The purpose of this study was to investigate, at the ultrastructural level, whether the ZI contains the anatomical substrata which could subserve the control of eye movements. METHODS: Blocks of tissue from the ZI of macaque monkeys were prepared for electron microscopy using standard techniques. Some of these animals were taken specifically for electron microscopy. Others had received injections of tracer substances and were prepared for electron microscopy subsequent to tracer visualization. RESULTS: Cell bodies of medium-large neurons were found in our preparations. They have large nucleoli and relatively small volumes of karyoplasm. Cell bodies and dendrites of all sizes have many synaptic contacts. Three types of synaptic profiles were found, designated Types 1, 2, and 3. Type 1 profiles are symmetrical and contact cell bodies and small dendrites. Type 2 profiles are thought to be presynaptic dendrites and may have symmetrical or asymmetrical contacts. Type 3 profiles are asymmetrical and primarily contact small dendrites. Many synapses contacted vesicle-containing profiles. In some cases, it was clear that these profiles participated in serial synapses on presumptive presynaptic dendrites. Other profiles appeared to be axoaxonic contacts. CONCLUSIONS: Afferent and efferent signals are likely to be modulated extensively within the ZI. Therefore, there needs to be complex interactions between neuronal elements of the ZI and its afferents. This study demonstrates that this nucleus possesses the structural substrata to subserve diverse roles, such as the gating of saccadic eye movements. PMID- 9335474 TI - Human facial muscles: dimensions, motor endplate distribution, and presence of muscle fibers with multiple motor endplates. AB - BACKGROUND: Extrafusal muscle fibers of human striated skeletal muscles are known to have a uniform innervation pattern. Motor endplates (MEP) of the "en plaque" type are located near the center of muscle fibers and distributed within the muscles in a narrow band. The aim of this study was to evaluate the innervation pattern of human facial muscles and compare it with that of skeletal muscles. METHODS: Ten facial muscles from 11 human cadavers were dissected, the nerve entrance points located, and the dimensions measured. All muscles were stained in toto for MEPs using Acetylcholinesterase (AChE) and examined under the microscope to determine their location. Single muscle fibers were teased to evaluate the stained MEPs. RESULTS: The length of the different facial muscles varied from 29 to 65 mm, which correlated to the length of the corresponding muscle fibers. MEP zones were found on the muscles in the immediate vicinity of the nerves' entrance points and located eccentrically. Numbers and locations varied from muscle to muscle. Three MEP zone distribution patterns were differentiated: numerous small MEP zones were evenly spread over the muscle, a predominant MEP zone and two to three small zones were spread at random, and two to four MEP zones of equal size were randomly scattered. One MEP of the "en plaque" type was found in 73.8% of the muscle fibers and two to five MEPs were found in 26.2%. The distances between the multiple MEPs on one muscle fiber varied from 10 to 500 microm. CONCLUSIONS: This study suggests that facial muscles differ from skeletal muscles regarding distribution and number of MEPs. The eccentric location of MEP zones and multiple MEPs suggests there is an independent mechanism of neural regulation in the facial muscle system. PMID- 9335475 TI - Morphology of the human internal vertebral venous plexus: a cadaver study after intravenous Araldite CY 221 injection. AB - Reviewing the literature on the vascular anatomy of the spinal epidural space, it appeared that the knowledge of the internal vertebral venous plexus is limited. Injection studies of the entire internal vertebral venous plexus after application of modern techniques, to the best of our knowledge, have never been performed. Based on the clinical importance of these structures, it was decided to study the human vertebral venous system after Araldite CY 221 injection, in order to update the morphological characteristics of the internal vertebral venous system. The vertebral venous systems of ten fresh human cadavers, between 64 and 93 years of age, were injected with Araldite CY 221 mixture. All cadavers were dissected and the posterior and anterior internal vertebral venous plexuses were studied in detail. The anterior part of the internal vertebral venous plexus is fairly constant. On the contrary, the posterior internal vertebral venous plexus showed a striking segmental and interindividual variability. In the thoracic area, two types of traversing veins are observed. Both types show a somewhat symmetrical "inversed V" configuration. No anatomical valves were observed. Nevertheless, anterograde flushing (via the femoral veins) of the vertebral venous system appeared to proceed much faster than retrograde flushing (via the superior vena cava). The classical picture of the internal vertebral venous plexus appears a simplification of the actual situation. Especially in the posterior part, segmental and interindividual differences are prominent. The preferential direction of the flow during flushing suggests the presence of functional valves, which are probably located in the thoracic part of the posterior internal vertebral venous plexus, resulting from the typical shape of the veins in this area. This might explain the difficulties with imaging of the posterior part of the internal vertebral venous plexus in vitro as well as in vivo. Further study is needed to determine whether the configuration of the posterior internal vertebral venous plexus in younger individuals is different, compared with the presently studied aged subjects. PMID- 9335476 TI - Nobel panel rewards prion theory after years of heated debate. PMID- 9335477 TI - Plagiarism claims turn spotlight on US Navy's misconduct policy. PMID- 9335478 TI - NCI apologizes for fallout study delay. PMID- 9335479 TI - Asia-Pacific forum backs life science network. PMID- 9335480 TI - Head of Third World biotechnology centre in Delhi resigns. PMID- 9335481 TI - Paul quits AIDS office to return to the lab. PMID- 9335482 TI - Monkey business in space. PMID- 9335484 TI - Definition of a gene. PMID- 9335483 TI - Bet you didn't know that. PMID- 9335485 TI - Duplications in nomenclature. PMID- 9335486 TI - Kainate receptors. Finding homes at synapses. PMID- 9335487 TI - Developmental biology. Straight and wiggly affinities. PMID- 9335488 TI - Chemotaxis. On the crest of a spiral wave. PMID- 9335489 TI - Signal transduction. Feedback from inhibitory SMADs. PMID- 9335490 TI - Telomerase. Cancer and the knockout mouse. PMID- 9335491 TI - Systemic signalling in gene silencing. PMID- 9335492 TI - A pandemic warning? PMID- 9335493 TI - DNA fingerprinting from single cells. PMID- 9335494 TI - Molecular motors: structural adaptations to cellular functions. AB - Molecular motors are protein machines whose directed movement along cytoskeletal filaments is driven by ATP hydrolysis. Eukaryotic cells contain motors that help to transport organelles to their correct cellular locations and to establish and alter cellular morphology during cell locomotion and division. The best-studied motors, myosin from skeletal muscle and conventional kinesin from brain, are remarkably similar in structure, yet have very different functions. These differences can be understood in terms of the 'duty ratio', the fraction of the time that a motor is attached to its filament. Differences in duty ratio can explain the diversity of structures, speeds and oligomerization states of members of the large kinesin, myosin and dynein families of motors. PMID- 9335495 TI - Bending and buckling of carbon nanotubes under large strain. AB - The curling of a graphitic sheet to form carbon nanotubes produces a class of materials that seem to have extraordinary electrical and mechanical properties. In particular, the high elastic modulus of the graphite sheets means that the nanotubes might be stiffer and stronger than any other known material, with beneficial consequences for their application in composite bulk materials and as individual elements of nanometre-scale devices and sensors. The mechanical properties are predicted to be sensitive to details of their structure and to the presence of defects, which means that measurements on individual nanotubes are essential to establish these properties. Here we show that multiwalled carbon nanotubes can be bent repeatedly through large angles using the tip of an atomic force microscope, without undergoing catastrophic failure. We observe a range of responses to this high-strain deformation, which together suggest that nanotubes are remarkably flexible and resilient. PMID- 9335496 TI - Convergent total synthesis of a tumour-associated mucin motif. AB - Synthetic glycoconjugates that mimic cell-surface tumour antigens (glycolipids or glycoproteins with unusual carbohydrate structural motifs) have been shown to trigger humoral responses in murine and human immune systems. This raises the exciting possibility of inducing active immunity with fully synthetic carbohydrate vaccines, particularly if vaccine compounds can be synthesized that resemble the surface environment of transformed cells even more closely. Glycopeptides seem particularly suitable for this purpose. In contrast to most glycolipids and the carbohydrates themselves, glycopeptides bind to major histocompatibility complex molecules, and, in favourable cases, can stimulate T cells and lead to the expression of receptors that recognize the carbohydrate part of a glycopeptide with high specificity. The preparation of glycopeptides and glycoproteins remains, however, a difficult challenge: earlier synthesis methods have been inefficient, and established cloning approaches that allow engineering of global glycopatterns produce only heterogeneous glycoproteins. Here we report an efficient strategy of the synthesis of tumour-associated mucin glycopeptides with clustered trisaccharide glycodomains corresponding to the (2,6)-sialyl T antigen. Our approach involves construction of the complete glycodomain in the first stage, followed by convergent coupling to amino acid residues and subsequent incorporation of the glycosyl amino acid units into a peptide chain. This general strategy allows the assembly of molecules in which selected glycoforms can be incorporated at any desired position of the peptide chain. The resultant fully synthetic O-linked glycopeptide clusters are the closest homogeneous mimics of cell-surface mucins at present available, and so are promising compounds for the development of anticancer vaccines. PMID- 9335497 TI - Fitness loss and germline mutations in barn swallows breeding in Chernobyl. AB - The severe nuclear accident at Chernobyl in 1986 resulted in the worst reported accidental exposure of radioactive material to free-living organisms. Short-term effects on human populations inhabiting polluted areas include increased incidence of thyroid cancer, infant leukaemia, and congenital malformations in newborns. Two recent studies have reported, although with some controversy, that germline mutation rates were increased in humans and voles living close to Chernobyl, but little is known about the viability of the organisms affected. Here we report an increased frequency of partial albinism, a morphological aberration associated with a loss of fitness, among barn swallows, Hirundo rustica, breeding close to Chernobyl. Heritability estimates indicate that mutations causing albinism were at least partly of germline origin. Furthermore, evidence for an increased germline mutation rate was obtained from segregation analysis at two hypervariable microsatellite loci, indicating that mutation events in barn swallows from Chernobyl were two- to tenfold higher than in birds from control areas in Ukraine and Italy. PMID- 9335498 TI - How the brain learns to see objects and faces in an impoverished context. AB - A degraded image of an object or face, which appears meaningless when seen for the first time, is easily recognizable after viewing an undegraded version of the same image. The neural mechanisms by which this form of rapid perceptual learning facilitates perception are not well understood. Psychological theory suggests the involvement of systems for processing stimulus attributes, spatial attention and feature binding, as well as those involved in visual imagery. Here we investigate where and how this rapid perceptual learning is expressed in the human brain by using functional neuroimaging to measure brain activity during exposure to degraded images before and after exposure to the corresponding undegraded versions. Perceptual learning of faces or objects enhanced the activity of inferior temporal regions known to be involved in face and object recognition respectively. In addition, both face and object learning led to increased activity in medial and lateral parietal regions that have been implicated in attention and visual imagery. We observed a strong coupling between the temporal face area and the medial parietal cortex when, and only when, faces were perceived. This suggests that perceptual learning involves direct interactions between areas involved in face recognition and those involved in spatial attention, feature binding and memory recall. PMID- 9335500 TI - Amyloidogenic role of cytokine TGF-beta1 in transgenic mice and in Alzheimer's disease. AB - Deposition of amyoid-beta peptide in the central nervous system is a hallmark of Alzheimer's disease and a possible cause of neurodegeneration. The factors that initiate or promote deposition of amyloid-beta peptide are not known. The transforming growth factor TGF-beta1 plays a central role in the response of the brain to injury, and increased TGF-beta1 has been found in the central nervous system of patients with Alzheimer's disease. Here we report that TGF-beta1 induces amyloid-beta deposition in cerebral blood vessels and meninges of aged transgenic mice overexpressing this cytokine from astrocytes. Co-expression of TGF-beta1 in transgenic mice overexpressing amyloid-precursor protein, which develop Alzheimer's like pathology, accelerated the deposition of amyloid-beta peptide. More TGF-beta1 messenger RNA was present in post-mortem brain tissue of Alzheimer's patients than in controls, the levels correlating strongly with amyloid-beta deposition in the damaged cerebral blood vessels of patients with cerebral amyloid angiopathy. These results indicate that overexpression of TGF beta1 may initiate or promote amyloidogenesis in Alzheimer's disease and in experimental models and so may be a risk factor for developing Alzheimer's disease. PMID- 9335499 TI - A hippocampal GluR5 kainate receptor regulating inhibitory synaptic transmission. AB - The principal excitatory neurotransmitter in the vertebrate central nervous system, L-glutamate, acts on three classes of ionotripic glutamate receptors, named after the agonists AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxalole-4 propionic acid), NMDA (N-methyl-D-aspartate) and kainate. The development of selective pharmacological agents has led to a detailed understanding of the physiological and pathological roles of AMPA and NMDA receptors. In contrast, the lack of selective kainate receptor ligands has greatly hindered progress in understanding the roles of kainate receptors. Here we describe the effects of a potent and selective agonist, ATPA ((RS)-2-amino-3-(3-hydroxy-5-tert butylisoxazol-4-yl)propanoic acid) and a selective antagonist, LY294486 ((3SR, 4aRS, 6SR, 8aRS)-6-((((1H-tetrazol-5-yl) methyl)oxy)methyl)-1, 2, 3, 4, 4a, 5, 6, 7, 8, 8a-decahydroisoquinoline-3-carboxylic acid), of the GluR5 subtype of kainate receptor. We have used these agents to show that kainate receptors, comprised of or containing GluR5 subunits, regulate synaptic inhibition in the hippocampus, an action that could contribute to the epileptogenic effects of kainate. PMID- 9335501 TI - An epithelial serine protease activates the amiloride-sensitive sodium channel. AB - Sodium balance, and ultimately blood pressure and extracellular fluid volume, is maintained by precise regulation of the activity of the epithelial sodium channel (ENaC). In a Xenopus kidney epithelial cell line (A6), exposure of the apical membrane to the protease inhibitor aprotinin reduces transepithelial sodium transport. Sodium-channel activity can be restored by subsequent exposure to the nonspecific protease trypsin. Using A6 cells and a functional complementation assay to detect increases in ENaC activity, we have cloned a 329-residue protein belonging to the serine protease family. We show that coexpression of this protein with ENaC in Xenopus oocytes increases the activity of the sodium channel by two- to threefold. This channel-activating protease (CAP1) is expressed in kidney, gut, lung, skin and ovary. Sequence analysis predicts that CAP1 is a secreted and/or glycosylphosphatidylinositol-anchored protein: ENaC activity would thus be regulated by the activity of a protease expressed at the surface of the same cell. This previously undiscovered mechanism for autocrine regulation may apply to other ion channels, in particular to members of the ENaC family that are present in neurons and epithelial cells. PMID- 9335502 TI - Protection from obesity-induced insulin resistance in mice lacking TNF-alpha function. AB - Obesity is highly associated with insulin resistance and is the biggest risk factor for non-insulin-dependent diabetes mellitus. The molecular basis of this common syndrome, however, is poorly understood. It has been suggested that tumour necrosis factor (TNF)-alpha is a candidate mediator of insulin resistance in obesity, as it is overexpressed in the adipose tissues of rodents and humans and it blocks the action of insulin in cultured cells and whole animals. To investigate the role of TNF-alpha in obesity and insulin resistance, we have generated obese mice with a targeted null mutation in the gene encoding TNF-alpha and those encoding the two receptors for TNF-alpha. The absence of TNF-alpha resulted in significantly improved insulin sensitivity in both diet-induced obesity and that resulting for the ob/ob model of obesity. The TNFalpha-deficient obese mice had lower levels of circulating free fatty acids, and were protected from the obesity-related reduction in the insulin receptor signalling in muscle and fat tissues. These results indicate that TNF-alpha is an important mediator of insulin resistance in obesity through its effects on several important sites of insulin action. PMID- 9335503 TI - Control of compartmental affinity boundaries by hedgehog. AB - In Drosophila, each segmental primordium is subdivided into two cell populations, the anterior (A) and posterior (P) compartments by the selective activity of the transcription factor Engrailed (En) in P cells. Under En control, P cells secrete, but cannot respond to, the signalling protein Hedgehog (Hh). In contrast, and by default, A cells are programmed to respond to Hh by expressing other signalling molecules, such as Decapentaplegic (Dpp) and Wingless (Wg), which organize growth and patterning in both compartments. Cells of the A and P compartments do not intermix, apparently as a consequence of their having distinct cell affinities that cause them to maximize contact with cells of the same compartment, while minimizing contact with cells from the other compartment. This failure to mix has previously been ascribed to an autonomous and direct role for En in specifying a P, as opposed to an A, cell affinity. However, an alternative hypothesis is that Hh secreted by P cells induces A cells to acquire a distinct cell affinity, ensuring that a stable 'affinity boundary' forms wherever P and A cells meet. Here we show that the affinity boundary that segregates A and P cells into adjacent but immiscible cell populations is to a large extent a consequence of local Hh signalling, rather than a reflection of an intrinsic affinity difference between A and P cells. PMID- 9335504 TI - Opposing BMP and EGF signalling pathways converge on the TGF-beta family mediator Smad1. AB - The growth factor TGF-beta, bone morphogenetic proteins (BMPs) and related factors regulate cell proliferation, differentiation and apoptosis, controlling the development and maintenance of most tissues. Their signals are transmitted through the phosphorylation of the tumour-suppressor SMAD proteins by receptor protein serine/threonine kinases (RS/TKs), leading to the nuclear accumulation and transcriptional activity of SMAD proteins. Here we report that Smadl, which mediates BMP signals, is also a target of mitogenic growth-factor signalling through epidermal growth factor and hepatocyte growth factor receptor protein tyrosine kinases (RTKs). Phosphorylation occurs at specific serines within the region linking the inhibitory and effector domains of Smad1, and is catalysed by the Erk family of mitogen-activated protein kinases. In contrast to the BMP stimulated phosphorylation of Smad1, which affects carboxy-terminal serines and induces nuclear accumulation of Smad1, Erk-mediated phosphorylation specifically inhibits the nuclear accumulation of Smad1. Thus, Smadl receives opposing regulatory inputs through RTKs and RS/TKs, and it is this balance that determines the level of Smad1 activity in the nucleus, and so possibly the role of Smad1 in the control of cell fate. PMID- 9335505 TI - Smad6 inhibits signalling by the TGF-beta superfamily. AB - SMAD proteins have been identified as signalling mediators of the TGF-beta superfamily, which is involved in a range of biological activities including cell growth, morphogenesis, development and immune responses. Smad1, Smad2, Smad3 and Smad5 are ligand-specific: Smadl and Smad5 transduce signals from bone morphogenetic proteins, and Smad2 and Smad3 mediate signalling by TGF-beta and activin, whereas Smad4 acts as a common signalling component. For example, Smad2 is phosphorylated by the TGF-beta type I receptor upon ligand binding, forms a heteromer with Smad4, and then translocates into the nucleus where it activates transcription. Here we report the isolation of Smad6 in the mouse. Smad6 is quite different in structure from the other SMAD proteins, and forms stable associations with type I receptors. Smad6 interferes with the phosphorylation of Smad2 and the subsequent heteromerization with Smad4, but does not inhibit the activity of Smad3. Smad6 also inhibits the phosphorylation of Smad1 that is induced by the bone morphogenetic protein type IB receptor. These data indicate that signals of the TGF-beta superfamily are regulated both positively and negatively by members of the SMAD family. PMID- 9335506 TI - Daughters against dpp modulates dpp organizing activity in Drosophila wing development. AB - The family of TGF-beta signalling molecules play inductive roles in various developmental contexts. One member of this family, Drosophila Decapentaplegic (Dpp) serves as a morphogen that patterns both the embryo and adult. We have now isolated a gene, Daughters against dpp (Dad), whose transcription is induced by Dpp. Dad shares weak homology with Drosophila Mad (Mothers against dpp), a protein required for transduction of Dpp signals. In contrast to Mad or the activated Dpp receptor, whose overexpression hyperactivates the Dpp signalling pathway, overexpression of Dad blocks Dpp activity. Expression of Dad together with either Mad or the activated receptor rescues phenotypic defects induced by each protein alone. Dad can also antagonize the activity of a vertebrate homologue of Dpp, bone morphogenetic protein, as evidenced by induction of dorsal or neural fate following overexpression in Xenopus embryos. We conclude that the pattern-organizing mechanism governed by Dpp involves a negative-feedback circuit in which Dpp induces expression of its own antagonist, Dad. This feedback loop appears to be conserved in vertebrate development. PMID- 9335507 TI - Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling. AB - TGF-beta signals from the membrane to the nucleus through serine/threonine kinase receptors and their downstream effectors, termed SMAD proteins. The activated TGF beta receptor induces phosphorylation of two such proteins, Smad2 and Smad3, which form hetero-oligomeric complex(es) with Smad4/DPC4 that translocate to the nucleus, where they then regulate transcriptional responses. However, the mechanisms by which the intracellular signals of TGF-beta are switched off are unclear. Here we report the identification of Smad7, which is related to Smad6. Transfection of Smad7 blocks responses mediated by TGF-beta in mammalian cells, and injection of Smad7 RNA into Xenopus embryos blocks activin/TGF-beta signalling. Smad7 associates stably with the TGF-beta receptor complex, but is not phosphorylated upon TGF-beta stimulation. TGFbeta-mediated phosphorylation of Smad2 and Smad3 is inhibited by Smad7, indicating that the antagonistic effect of Smad7 is exerted at this important regulatory step. TGF-beta rapidly induces expression of Smad7 mRNA, suggesting that Smad7 may participate in a negative feedback loop to control TGF-beta responses. PMID- 9335508 TI - Survival of FimH-expressing enterobacteria in macrophages relies on glycolipid traffic. AB - Strains of Escherichia coli persist within the human gut as normal commensals, but are frequent pathogens and can cause recurrent infection. Here we show that, in contrast to E. coli subjected to opsonic interactions stimulated by the host's immune response, E. coli that bind to the macrophage surface exclusively through the bacterial lectin FimH can survive inside the cell following phagocytosis. This viability is largely due to the attenuation of intracellular free-radical release and of phagosome acidification during FimH-mediated internalization, both of which are triggered by antibody-mediated internalization. This different processing of non-opsonized bacteria is supported by morphological evidence of tight-fitting phagosomes compared with looser, antibody-mediated phagosomes. We propose that non-opsonized FimH-expressing E. coli co-opt internalization of lipid-rich microdomains following binding to the FimH receptor, the glycosylphosphatidylinositol-linked protein CD48, because (1) the sterol-binding agents filipin, nystatin and methyl beta-cyclodextrin specifically block FimH mediated internalization; (2) CD48 and the protein caveolin both accumulate on macrophage membranes surrounding bacteria; and (3) antibodies against CD48 inhibit FimH-mediated internalization. Our findings bring the traditionally extracellular E. coli into the realm of opportunistic intracellular parasitism and suggest how opportunistic infections with FimH-expressing enterobacteria could occur in a setting deprived of opsonizing antibodies. PMID- 9335510 TI - Introduction: taxoids and the management of breast cancer. PMID- 9335509 TI - Mitotic chromatin regulates phosphorylation of Stathmin/Op18. AB - Meiotic and mitotic spindles are required for the even segregation of duplicated chromosomes to the two daughter cells. The mechanism of spindle assembly is not fully understood, but two have been proposed that are not mutually exclusive. The 'search and capture' model suggests that dynamic microtubules become progressively captured and stabilized by the kinetochores on chromosomes, leading to spindle assembly. The 'local stabilization' model proposes that chromosomes change the state of the cytoplasm around them, making it more favourable to microtubule polymerization. It has been shown that Stathmin/Op18 inhibits microtubule polymerization in vitro by interaction with tubulin, and that overexpression in tissue culture cells of non-phosphorylatable mutants of Stathmin/Op18 prevents the assembly of mitotic spindles. We have used Xenopus egg extracts and magnetic chromatin beads to show that mitotic chromatin induces phosphorylation of Stathmin/Op18. We have also shown that Stathmin/Op18 is one of the factors regulated by mitotic chromatin that governs preferential microtubule growth around chromosomes during spindle assembly. PMID- 9335511 TI - The taxoids: same roots, different drugs. AB - The discovery and development of the taxoid class of antitumor compounds, most notably paclitaxel, originally extracted from the Pacific yew, and its semisynthetic analog docetaxel, represent significant advances in the treatment of patients with a variety of malignancies. Although paclitaxel and docetaxel have a similar chemical root, extensive research and clinical experience indicate that important biological and clinical differences exist between the two compounds. Although the mechanism by which they disrupt mitosis and cell replication is novel and unique to this class of compounds, there are small but important differences in the formation of the stable, nonfunctional microtubule bundles and in the affinity of the two compounds for binding sites. These differences may explain the lack of complete cross-resistance observed between docetaxel and paclitaxel in preclinical and clinical studies. The two taxoids also exhibit slightly different toxicity and clinical efficacy profiles. Docetaxel-induced adverse events have been shown to occur most frequently in patients with impaired liver function, and a reduction in dosage in patients with raised levels of transaminases and alkaline phosphatase is recommended to improve the tolerability of the drug in these patients. The incidence and severity of problematic adverse effects such as hypersensitivity, skin reactions, and a cumulative fluid retention syndrome are minimized with the routine administration of a 3- to 5-day corticosteroid-based premedication regimen with each docetaxel infusion. The results of ongoing studies of taxoid-based monotherapies and combination regimens will further optimize the benefits achievable with these drugs, and new preclinical findings also may lead to new clinical uses for these and future drugs of the taxoid class. PMID- 9335512 TI - Docetaxel as single-agent therapy in metastatic breast cancer: clinical efficacy. AB - The potential importance of docetaxel as a chemotherapeutic agent for the treatment of metastatic breast cancer has been documented in approximately 575 patients enrolled in phase II clinical studies in North America, Europe, and Japan. First-line treatment with docetaxel 100 mg/m2 intravenously over 1 hour every 3 weeks produced objective responses in 59% of patients (intent-to-treat). In patients with anthracycline-resistant disease (disease that responded initially but then progressed or relapsed during treatment with an anthracycline), an overall intent-to-treat response rate of 41% was achieved. A 37% response rate was observed in anthracycline-refractory patients (those who had never achieved an objective response to anthracycline-based chemotherapy). Febrile neutropenia is the major dose-limiting toxicity of docetaxel; the neutropenia nadir typically has an early onset and short duration, with rapid recovery of the neutrophil count. Other hematologic effects are rare. Skin and hypersensitivity reactions can be minimized by premedication with corticosteroids. Reversible fluid retention, related to the cumulative dose of docetaxel, is also lessened by corticosteroid prophylaxis. In summary, docetaxel has shown impressive antitumor activity in the treatment of metastatic breast cancer, including the challenging population of patients whose disease progressed during treatment with an anthracycline or anthracenedione. Most of the adverse events related to docetaxel use can be circumvented or ameliorated by careful patient selection and routine premedication with corticosteroids, and improved methods of managing side effects are being actively studied. Docetaxel is thus expected to play an increasingly prominent role in the management of patients with metastatic breast cancer. PMID- 9335513 TI - Docetaxel in combination chemotherapy for metastatic breast cancer. AB - Due to its novel mechanism of action, docetaxel has significant in vitro activity against a variety of solid tumors, including breast cancer. In phase II clinical trials, docetaxel 100 mg/m2 every 3 weeks has shown substantial single-agent activity in patients with both previously untreated and heavily pretreated metastatic breast cancer. As single-agent chemotherapy is rarely curative in this setting, docetaxel has been combined with other anticancer agents with proven efficacy against breast cancer (doxorubicin, vinorelbine, fluorouracil, cyclophosphamide, cisplatin, and doxorubicin plus cyclophosphamide) in an attempt to increase efficacy and prolong patient survival. The aims of these phase I dose finding studies were to define the maximum tolerated dose, the dose-limiting toxicity, the recommended dose, and the safety profile of each combination. All patients had previously untreated metastatic breast cancer. Each study followed a dose-escalation design; patients were initially assigned to the lowest dose level, with at least three patients treated at each level before enrolling patients at the next level. All patients received corticosteroid-based premedication, but granulocyte colony-stimulating factor was not routinely used. The docetaxel/doxorubicin and docetaxel/ vinorelbine studies have been recently completed; in both studies, the dose-limiting toxicity was neutropenia. Other adverse events were generally mild. No significant cardiotoxicity (with docetaxel/doxorubicin) or neurotoxicity (with docetaxel/vinorelbine) was observed, and no patients discontinued treatment because of fluid retention. Both combinations are well tolerated without granulocyte colony-stimulating factor support. The recommended dose for docetaxel/doxorubicin in phase II studies is either 75/50 or 60/60 mg/m2 administered on day 1 every 3 weeks; impressive response rates of 90% and 66%, respectively, were achieved with these dose levels. The recommended dose for docetaxel/vinorelbine is docetaxel 85 mg/m2 on day 1 and vinorelbine 20 mg/m2 on days 1 and 5, repeated every 3 weeks; the efficacy of this combination is still being evaluated, but preliminary results are promising. For both combinations, the time to disease progression and median response duration were not available at the time of this report. Combination studies with fluorouracil, cisplatin, and cyclophosphamide, and a study of sequential administration with doxorubicin + cyclophosphamide, are ongoing. Interim results indicate that these docetaxel-based combinations have acceptable safety profiles and encouraging levels of antitumor activity. The full results of these studies will help to elucidate the potential contribution of docetaxel based combination chemotherapy to the management of metastatic breast cancer. PMID- 9335514 TI - Docetaxel and paclitaxel in breast cancer therapy: present status and future prospects. AB - The taxoids are important novel drugs in the treatment of patients with metastatic breast cancer, including those patients with extensive prior therapy and even patients with anthracycline-refractory disease. The emerging role of taxoid therapy and its optimal integration into new treatment strategies for patients with breast cancer is a major focus of active clinical and laboratory research. The incomplete clinical cross-resistance with anthracyclines observed thus far with both paclitaxel and docetaxel is particularly encouraging, as therapeutic options for patients after anthracycline failure are few and often of limited clinical value. Results of ongoing randomized multicenter trials addressing important clinical questions of optimal dose level, infusion duration, definition of worthwhile combinations, and comparative efficacy and tolerability of taxoid monotherapy or combination therapy with other standard regimens will, in the near future, provide guidelines for the best use of these drugs. Beyond the demonstrated single-agent activity of the taxoids in metastatic breast cancer, trials evaluating the taxoids in combination with various chemotherapeutic agents and monoclonal antibodies are in progress. Pharmacokinetic evaluation of the taxoids as monotherapy and in combination should help to define optimal and safe drug delivery. Clinical and laboratory studies will ultimately define clinically relevant mechanisms of resistance to this class of drugs, and may thus direct the development of rational strategies to overcome resistance and/or to guide analogue development. Finally, follow-up of women participating in present randomized adjuvant trials evaluating the taxoids will determine the potential impact of these important agents on the curability of early stage breast cancer. PMID- 9335515 TI - The development of docetaxel (Taxotere) in non-small cell lung cancer. PMID- 9335516 TI - Combination treatment with docetaxel (Taxotere) and platinum compounds for non small cell lung cancer. AB - There is a strong rationale for combining docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) and platinum compounds for use in the treatment of non small cell lung cancer (NSCLC). The compounds are active as single agents and have no overlapping toxicity profiles. The first dose-finding study reported a response rate of 46%, and recommended doses of 75 mg/m2 for docetaxel and 75 to 100 mg/m2 for cisplatin, given concomitantly every 3 weeks. Three other studies confirmed a response rate of 33% to 48% in previously untreated patients. Dose limiting but rapidly reversible neutropenia was observed. A European trial has investigated the use of alternating cycles of docetaxel 100 mg/m2 and cisplatin 120 mg/m2 in patients with previously untreated metastatic or nonresectable NSCLC. Preliminary results indicate that the overall response rate (33% of 39 evaluable patients) is similar to that achieved with docetaxel alone. A second trial is investigating alternating therapy using docetaxel 100 mg/m2 and a combination of cisplatin 100 mg/m2 and vinorelbine 30 mg/m2. Response rates are higher (49%) and toxicities, especially hematologic toxicities, are lower than in the first study. A third trial is investigating sequential treatment using docetaxel 100 mg/m2, cisplatin 120 mg/m2, and vindesine 3 mg/m2. Preliminary results indicate a response rate of 30% among 25 patients enrolled thus far. Compared with monotherapy, docetaxel/platinum combinations appear to offer the potential for improved response rates in patients with NSCLC. The use of alternating schedules offers the further possible advantage of improving tolerability while retaining effective doses. Further studies are in progress or planned to elucidate optimal doses and schedules. PMID- 9335517 TI - Docetaxel (Taxotere) and vinorelbine in the treatment of non-small cell lung cancer. AB - The efficacy and safety of a combination of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) and vinorelbine were evaluated in two phase II studies including 46 and 39 chemotherapy-naive patients with non-small cell lung cancer, respectively. In the first study, vinorelbine 25 mg/m2 was given on day 1 and docetaxel 100 mg/m2 on day 2, with recombinant human granulocyte colony stimulating factor support from day 5 until day 12, every 3 weeks. In the second study, docetaxel 75 mg/m2 was given on day 1 and vinorelbine 20 to 25 mg/m2 on days 1 and 5 in a 3-week schedule. Grade 3/4 neutropenia was the most severe toxicity, occurring in 46% to 85% of patients, while febrile neutropenia was observed in 25% to 41% of patients. Two treatment-related deaths occurred in each study. Patients with a poor performance status were at an increased risk of infection. The objective response rates were 36.6% and 27%, respectively (mean, 32%). In two subsequent studies, the efficacy and safety of a triple-drug combination of docetaxel, vinorelbine, and cisplatin was evaluated. In the first study, 14 patients were treated with vinorelbine 25 mg/m2 and cisplatin 80 mg/m2 on day 1 and docetaxel 100 mg/m2 and vinorelbine 20 mg/m2 on day 8, every 3 weeks. In the second study, 46 patients received docetaxel 100 mg/m2 on day 1, cisplatin 100 mg/m2 and vinorelbine 30 mg/m2 on day 21, and vinorelbine 30 mg/m2 on days 28 and 35, every 6 weeks. The main toxicity was also grade 3/4 neutropenia (57% and 80% of patients, respectively). Febrile neutropenia developed in 50% and 15% of the patients in the first and second study, respectively. The overall response rates were 33% and 39%, respectively. It is concluded that combination therapy with docetaxel/ vinorelbine or with docetaxel/vinorelbine/cisplatin has antitumor activity in the treatment of non small cell lung cancer. Granulocytopenia and febrile neutropenia are the most severe toxicities that limit the usefulness of these regimens. PMID- 9335518 TI - Docetaxel (Taxotere) and vinorelbine in the treatment of advanced non-small cell lung cancer: preliminary results of a phase I/II trial. AB - We undertook a phase I/II study of the combination of docetaxel (Taxotere; Rhone Poulenc Rorer, Antony, France) and vinorelbine in the treatment of unresectable or metastatic non-small cell lung cancer (NSCLC). Nineteen patients with unresectable NSCLC received a combination of docetaxel 50 mg/m2 and vinorelbine 15 to 45 mg/m2 every 2 weeks. All patients received prophylactic granulocyte colony stimulating factor 5 microg/kg/d and corticosteroids. The incidence of hematologic toxicity with this dosing schedule was low; only 2.5% of treatment cycles were complicated by febrile neutropenia. The maximum tolerated dose has not been reached. It is concluded that using this schedule, a combination of docetaxel and vinorelbine can be administered in combination, together with granulocyte-colony stimulating factor, with a low risk of associated hematologic toxicity. Further dose escalation is needed to determine the maximum tolerated dose of the combination. A partial response was observed in five patients (26%; 95% confidence interval, 15% to 57%), and nine (47%) patients showed stable disease. Based on these preliminary results, the combination of these two drugs appears to have antitumor activity against NSCLC and warrants continued study. PMID- 9335519 TI - The development of docetaxel (Taxotere) in non-small cell lung cancer--docetaxel in new combinations and new schedules: an overview of ongoing and future developments. AB - Non-small cell lung cancer is the most common cause of cancer death in the western world. Non-small cell lung cancer is modestly sensitive to chemotherapy with a small survival benefit in locally advanced and metastatic disease. Newer agents such as docetaxel are yielding encouraging response rates both as single agents and in combination. A phase I/II study is in progress in our institution to determine the maximum tolerated dose and noncomparative efficacy of the combination of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France), ifosfamide, and cisplatin, with mesna and lenograstim support, in the treatment of patients with advanced non-small cell lung cancer. To date, nine patients have received 37 cycles of treatment at increasing dose levels (no intrapatient dose escalation). Treatment was administered to patients on an inpatient basis every 3 weeks, with lenograstim on days 3 to 10. Dose-limiting toxicity has not occurred at levels I to III (dose level III: docetaxel 75 mg/m2, cisplatin 75 mg/m2, and ifosfamide 3 g/m2). These preliminary results suggest that the combination of docetaxel, ifosfamide, and cisplatin, with lenograstim support, is well tolerated in the doses evaluated. Preliminary efficacy results show a response rate of 67% (six of nine patients). The study continues to determine the maximum tolerated dose of this regimen in preparation for a phase II evaluation. PMID- 9335521 TI - Clinical studies of docetaxel (Taxotere) and concomitant chest therapy. AB - The administration of concomitant chemoradiotherapy has been shown to increase local and regional control of non-small cell lung cancer. Docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) has single-agent activity in non-small cell lung cancer, as well as radiation-enhancing potential in preclinical studies. Therefore, its investigation with concomitant radiotherapy in the clinical setting is justified. Since the clinical interactions of docetaxel and concomitant chest radiotherapy have not been previously described, we initiated a phase I study with the goal of determining the maximum tolerated dose of docetaxel and the optimal schedule for its administration during a standard course of radiation therapy to the chest, in addition to defining the dose limiting toxicities of this regimen. This report describes the design and preliminary results of this study. PMID- 9335520 TI - Docetaxel (Taxotere) and gemcitabine in the treatment of non-small cell lung cancer: preliminary results. AB - A phase II study was performed to investigate the tolerance and efficacy of the combination of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) and gemcitabine in chemotherapy-naive patients with advanced non-small cell lung cancer (NSCLC). To date, 24 patients (five with stage IIIB and 19 with stage IV NSCLC) have been treated according to the protocol: gemcitabine 900 mg/m2 was administered on days 1 and 8 as a 30-minute infusion and docetaxel 100 mg/m2 was administered on day 8 as a 1-hour infusion after appropriate premedication. Granulocyte colony-stimulating factor 150 microg/m2 subcutaneously was given on days 9 to 15. Treatment was repeated every 3 weeks. Grade 3/4 granulocytopenia occurred in seven (29%) patients, and one (4%) of these patients developed febrile neutropenia. Grade 3/4 thrombocytopenia and anemia were observed in three (13%) and one (4%) patient, respectively. Grade 2 neurotoxicity and fatigue occurred in one (4%) patient each. Other toxicities were mild. There were no treatment-related deaths. Eight patients experienced a partial response (53.3%; 95% confidence interval, 28.1% to 78.6%), and stable and progressive disease were documented in two (13%) and five (33%) patients, respectively. The median delivered dose was 600 mg/m2/wk and 33 mg/m2/wk for gemcitabine and docetaxel, respectively. These preliminary data suggest that the docetaxel/gemcitabine combination has significant antitumor activity and is well tolerated in chemotherapy-naive patients with NSCLC. The study is ongoing. PMID- 9335522 TI - The role of docetaxel (Taxotere) as a single agent or in combination before local treatment of non-small cell lung cancer. AB - Neoadjuvant therapy in the treatment of stage IIIa/b non-small cell lung cancer (NSCLC) has the potential to reduce tumor size in patients whose tumors were previously inoperable. This report describes the design and status of an ongoing randomized, phase III study of docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) as neoadjuvant treatment in patients with stage IIIa/b NSCLC, as well as of two phase II studies of combination neoadjuvant therapies. A phase III, multicenter, international, randomized trial is in progress which compares docetaxel with no neoadjuvant chemotherapy in patients with histologically confirmed, previously untreated NSCLC with stage IIIa N2(T0-3) or T3 (N0-1) disease or stage IIIb disease that can be treated radically. Patients are assigned to receive three cycles of docetaxel or no neoadjuvant therapy. Definitive therapy is administered immediately or within 6 weeks of the third cycle of docetaxel. The primary objective of the study is median survival. Secondary end points include response rate, tumor resectability, survival after surgery and after curative-intent radiation, time to disease progression, and quality of life. To date, 110 of the planned 292 patients have been randomized. It is anticipated that the study will be completed in 1998. A phase I/II dose escalation study of the combination of docetaxel and cisplatin as neoadjuvant treatment of patients with stage IIIa T1-T3 NSCLC with N2 disease on mediastinoscopy is ongoing. The objectives of the study are to determine the clinical response to chemotherapy, the pathologic response after surgery, and survival. A phase II, multicenter, nonrandomized trial of the combination of docetaxel and carboplatin as neoadjuvant therapy in patients with stage IIIa N2 NSCLC is in progress. The primary objective of the study is to determine the response rate after chemotherapy. The results of both studies should be available by late 1997. The ultimate hope is that there is potential for neoadjuvant chemotherapy to provide a significant benefit for patients with advanced NSCLC. PMID- 9335523 TI - The nuclear-encoded PsbW protein subunit of photosystem II undergoes light induced proteolysis. AB - The repair of photoinhibitory damage to photosystem II involves the rapid degradation and turnover of the D1 reaction center subunit. Additional protein subunits which show a limited degradation at high light intensities are the complementary reaction center subunit, D2, and the two chlorophyll a binding proteins, CP 47 and CP 43. In this work, we provide the first evidence for light induced degradation of a nuclear-encoded subunit of photosystem II, the recently discovered PsbW protein. This 6.1 kDa protein is predicted to have a single membrane span and was found to be closely associated with the photosystem II reaction center. The degradation of the PsbW protein was demonstrated by photoinhibitory experiments, both in vitro, using thylakoid membranes and photosystem II core particles, and in vivo using leaf discs. The PsbW protein showed almost the same rate and extent of degradation as the D1 protein, and its degradation was more pronounced compared to the D2 and CP 43 proteins. The degradation of the PsbW protein was shown to share many mechanistic similarities with the more well characterized D1 protein degradation, such as oxygen dependence, sensitivity to serine protease inhibitors, and high light triggering while the actual degradation could readily occur in total darkness. The degradation of the PsbW protein was impaired by protein phosphorylation, although this protein was not itself phosphorylated. This impairment was correlated to the phosphorylation of the D1 protein which has been shown to block its degradation during photoinhibitory conditions. It is concluded that the PsbW protein is not degraded as a direct consequence of primary photodamage but due to a general destabilization of the photosystem II complex under conditions were the D1 protein becomes degraded in the absence of a sufficient repair system. The results are discussed in terms of a requirement for coordination between degradation and protein synthesis/integration during the repair process of photodamaged photosystem II reaction centers. PMID- 9335524 TI - The substrate binding site of human liver cytochrome P450 2C9: an NMR study. AB - Purified recombinant human liver cytochrome P450 2C9 was produced, from expression of the corresponding cDNA in yeast, in quantities large enough for UV visible and 1H NMR experiments. Its interaction with several substrates (tienilic acid and two derivatives, lauric acid and diclofenac) and with a specific inhibitor, sulfaphenazole, was studied by UV-visible and 1H NMR spectroscopy. At 27 degrees C, all those substrates led to an almost complete conversion of CYP 2C9 to high-spin (S = 5/2) CYP 2C9-substrate complexes characterized by a Soret peak at 390 nm; their KD values varied between 1 and 42 microM. On the contrary, sulfaphenazole led to a low-spin (S = 1/2) CYP 2C9 complex upon binding of its NH2 group to CYP 2C9 iron. Interactions of the five substrates with the enzyme were studied by paramagnetic relaxation effects of CYP 2C9-iron(III) on the 1H NMR spectrum of each substrate. Distances between the heme iron atom and substrate protons were calculated from the NMR data, and the orientation of the substrate relative to iron was determined from those distances. Finally, a model for substrate positioning in the CYP 2C9 active site was constructed by molecular modeling studies under the constraint of the iron-proton distances. It points out two structural characteristics for a compound to be selectively recognized by CYP 2C9: (i) the presence of an anionic site able to establish an ionic bond with a putative cationic residue of the protein and (ii) the presence of an hydrophobic zone between the substrate hydroxylation site and the anionic site. Sulfaphenazole was easily included in that model; its very high affinity for CYP 2C9 is due to a third structural feature, the presence of its NH2 function which binds to CYP 2C9 iron. PMID- 9335525 TI - Structural characterization of an analog of the major rate-determining disulfide folding intermediate of bovine pancreatic ribonuclease A. AB - The major rate-determining step in the oxidative regeneration of bovine pancreatic ribonuclease A (RNase A) proceeds through des-[40-95] RNase A, a three disulfide intermediate lacking the Cys40-Cys95 disulfide bond. An analog of this intermediate, [C40A, C95A] RNase A, has been characterized in terms of regular backbone structure and thermodynamic stability at pH 4.6. Nearly complete backbone 1H, 15N, and 13C resonances, and most 13Cbeta side-chain resonances have been assigned for the mutant RNase A using triple-resonance NMR data and a computer program, AUTOASSIGN, for automated analysis of resonance assignments. Comparisons of chemical shift data, 3J(1HN-1Halpha) coupling constants, and NOE data for the mutant and wild-type proteins reveal that the overall chain folds of the two proteins are very similar, with localized structural perturbations in the regions spatially adjacent to the mutation sites in [C40A, C95A] RNase A. More significantly, 1H/2H amide exchange and thermodynamic data reveal a global destabilization of the mutant protein characterized by a significant difference in the midpoint of the thermal transition curves (DeltaTm of 21.8 degrees C) and a significant increase in the slowest exchanging backbone amide 1H/2H exchange rates (10(2)-10(6)-fold faster in the hydrophobic core of [C40A, C95 A] RNase A). Comparisons of the entropy DeltaS degrees (T) and enthalpy DeltaH degrees (T) of unfolding between wild-type and [C40A, C95A] RNase A reveal that some of the global destabilization of the mutant protein arises from entropic and enthalpic changes in the folded state. Implications of these observations for understanding the role of des-[40-95] in the folding pathway of RNase A are discussed. PMID- 9335526 TI - High-resolution structure of the extracellular aspartic proteinase from Candida tropicalis yeast. AB - The crystal structure of the secreted aspartic proteinase from Candida tropicalis yeast (SAPT) has been determined to 1.8 A resolution. The classic aspartic proteinase bilobal structure and domain topology is conserved in SAPT, with the substrate binding cleft situated between the two domains. Structural comparisons made with pepsin indicate that insertions and deletions in the primary sequence modify the SAPT structure to create a more spacious substrate binding cleft with altered specificity. An unexpected tetrapeptide has been found to occupy binding sites S1'-S3', and this suggests the order of release of peptide products in the catalytic mechanism of these enzymes. Structural features are considered with regard to previous substrate specificity data. PMID- 9335527 TI - Implications of RNA structure on the annealing of a potent antisense RNA directed against the human immunodeficiency virus type 1. AB - Antisense RNA-mediated regulation in bacterial systems is related to the kinetics of RNA-RNA annealing in vitro. Here, we investigated the secondary structure of alphaY69, an effective HIV-directed antisense RNA in human cells. Purified RNA preparations contain a single conformer. The global structure was identified by a cleavage experiment under native conditions using a short complementary oligonucleotide and RNase H. Structural analyses indicate a three-domain structure of alphaY69 consisting of two stem-loop elements connected by a seven nucleotide single-stranded hinge region. Kinetic data suggest that the formation of base pairs between a CGC triplet of alphaY69 and its target RNA is essential for fast annealing. The complementary sequence stretch of the target folds into a high-energy secondary structure. The relationship between modifications in structural elements of alphaY69 and the annealing kinetics suggested that rate limiting steps of the annealing involve a single site of alphaY69 and do not involve its 5' or 3'-end. Further, the data indicate that both initial base specific interactions and duplex formation are dependent on the CGC triplet of the central region of alphaY69. This mechanism represents a specific and efficient way of RNA-RNA annealing that is initiated by the interaction of unstructured RNA regions. PMID- 9335528 TI - RNA recognition by a bent alpha-helix regulates transcriptional antitermination in phage lambda. AB - A novel RNA recognition motif is characterized in an arginine-rich peptide. The motif, derived from lambda transcriptional antitermination protein N, regulates an RNA-directed genetic switch. Its characterization by multidimensional nuclear magnetic resonance (NMR) demonstrates specific RNA-dependent folding of N- and C terminal recognition helices separated by a central bend. The biological importance of the bent alpha-helix is demonstrated by mutagenesis: binding is blocked by substitutions in the N peptide or its target (the boxB RNA hairpin) associated in vivo with loss of transcriptional antitermination activity. Although arginine side chains are essential, the peptide is also anchored to boxB by specific nonpolar contacts. An alanine in the N-terminal helix docks in the major groove of the RNA stem whereas a tryptophan in the C-terminal helix stacks against a purine in the RNA loop. At these positions all 19 possible amino acid substitutions have been constructed by peptide synthesis; each impairs binding to boxB. The pattern of allowed and disallowed substitutions is in accord with the results of random-cassette mutagenesis in vivo. The helix-bend-helix motif rationalizes genetic analysis of N-dependent transcriptional antitermination and extends the structural repertoire of arginine-rich domains observed among mammalian immunodeficiency viruses. PMID- 9335529 TI - Factors which stabilize the methylamine dehydrogenase-amicyanin electron transfer protein complex revealed by site-directed mutagenesis. AB - Methylamine dehydrogenase (MADH) and amicyanin form a physiologic complex within which electrons are transferred from the tryptophan tryptophylquinone (TTQ) cofactor of MADH to the type 1 copper of amicyanin. Interactions responsible for complex formation may be inferred from the crystal structures of complexes of these proteins. Site-directed mutagenesis has been performed to probe the roles of specific amino acid residues of amicyanin in stabilizing the MADH-amicyanin complex and determining the observed ionic strength dependence of complex formation. Conversion of Phe97 to Glu severely disrupted binding, establishing the importance of hydrophobic interactions involving this residue. Conversion of Arg99 to either Asp or to Leu increased the Kd for complex formation by 2 orders of magnitude at low ionic strength, establishing the importance of ionic interactions which were inferred from the crystal structure involving Arg99. Conversion of Lys68 to Ala did not disrupt binding at low ionic strength, but it did greatly diminish the observed ionic strength dependence of complex formation that is seen with wild-type amicyanin. These results demonstrate that the physiologic interaction between MADH and amicyanin is stabilized by a combination of ionic and van der Waals interactions and that individual amino acid residues on the protein surface are able to dictate specific interactions between these soluble redox proteins. These results also indicate that the orientation of MADH and amicyanin when they react with each other in solution is the same as the orientation of the proteins which is seen in the structure of the crystallized protein complex. PMID- 9335530 TI - A response regulatory protein with the site of phosphorylation blocked by an arginine interaction: crystal structure of Spo0F from Bacillus subtilis. AB - Spo0F is a secondary messenger in the "two-component" system controlling the sporulation of Bacillus subtilis. Spo0F, like the chemotaxis protein CheY, is a single-domain protein homologous to the N-terminal activator domain of the response regulators. We recently reported the crystal structure of a phosphatase resistant mutant Y13S of Spo0F with Ca2+ bound in the active site. The crystal structure of wild-type Spo0F in the absence of a metal ion is presented here. A comparison of the two structures reveals that the cation induces significant changes in the active site. In the present wild-type structure, the carboxylate of Asp11 points away from the center of the active site, whereas when coordinated to the Ca2+, as in the earlier structure, it points toward the active site. In addition, Asp54, the site of phosphorylation, is blocked by a salt bridge interaction of an Arg side chain from a neighboring molecule. From fluorescence quenching studies with Spo0F Y13W, we found that only the amino acid Arg binds to Spo0F in a saturable manner (Kd = 15 mM). This observation suggests that a small molecule with a shape complementary to the active site and having a guanidinium group might inhibit phosphotransfer between response regulators and their cognate histidine kinases. PMID- 9335531 TI - Binding of 2,6- and 2,7-dihydroxynaphthalene to wild-type and E-B13Q insulins: dynamic, equilibrium, and molecular modeling investigations. AB - The binding of phenolic ligands to the insulin hexamer occurs as a cooperative allosteric process. Investigations of the allosteric mechanism from this laboratory resulted in the postulation of a model consisting of a three-state conformational equilibrium and the derivation of a mathematical expression to describe the insulin system. The proposed mechanism involves allosteric transitions among two states of high symmetry, designated T3T3' (a low affinity state) and R3R3' (a high affinity state), and a third state of lower symmetry, designated T3oR3o (a state of mixed low and high affinities). To further characterize this mechanism, we present rapid kinetic fluorescence studies, equilibrium binding isotherms, and molecular modeling investigations for the Co(II)-substituted wild-type and E-B13Q mutant hexamers. These studies show that the measured on and off rates (kon and koff) for the binding of the allosteric ligands 2,6- and 2,7-dihydroxynaphthalene provide an independent measure of the dissociation constant for binding to the T3oR3o conformation (KRo). These constants are in agreement with the value obtained by computer fitting of the equilibrium binding isotherms to the quantitative allosteric mechanism. We analyze the structural differences between the T3oR3o and R6 phenolic binding sites and predict the structures of the T3oR3o-2,6-DHN and R6-2, 6-DHN complexes by 3-D molecular modeling. Assignment of H-bonding of the first hydroxyl group to CysA6 and CysA11 has been supported by stacking interactions analogous to phenol using 1H-NMR. H-bonding of the second hydroxyl group of 2,6-DHN to the GluB13 carboxylate side chains is predicted by molecular modeling and is supported by a reduction of affinity for Ca2+, which is postulated to bind to the GluB13 side chains. PMID- 9335532 TI - Half-site reactivity, negative cooperativity, and positive cooperativity: quantitative considerations of a plausible model. AB - The nature of cooperative allosteric interactions has been the source of controversy since the ground-breaking studies of oxygen binding to hemoglobin. Until recently, quantitative examples of a model based on the inherent symmetry and asymmetry of oligomeric proteins have been lacking. This laboratory has used the phenolic ligand binding characteristics of the insulin hexamer to develop the first quantitative model for a symmetry-asymmetry-based cooperativity mechanism. The insulin hexamer possesses positive and negative heterotropic and homotropic interactions involving two classes of sites. In this study, we explore the effects of heterotropic interactions between these sites. We show that application of the pairwise structural asymmetry theory of Seydoux, Malhotra, and Bernhard (SMB) gives excellent agreement between the ligand binding behavior and X-ray crystal structure data. Furthermore, by comparing experimental data with computer simulations, we show that the insulin hexamer can be described by a three-state SMB model involving two positive homotropic cooperative transitions linked by a negative homotropic interaction. The first transition, T3T3' right harpoon over left harpoon T3oR3o, with allosteric constant LoA = [T3T3']/[T3oR3o] and ligand dissociation constant KRo consists of a positive cooperative change from high to low symmetry that results in "half-site reactivity". The second transition, T3oR3o right harpoon over left harpoon R3R3', with allosteric constant LoB = [T3oR3o]/[R3R3'] and ligand dissociation constant KR is a change from low to high symmetry, which is also a positive cooperative process. Treatment of the two transitions as concerted and interconnected processes allows derivation of an equation for the fraction of R-state. Using this equation, the effects of changes in the four physical parameters, LoA, LoB, KR, and KRo, on the ligand binding properties of the insulin hexamer are quantitatively described. PMID- 9335533 TI - Heparan sulfate proteoglycan-mediated uptake of apolipoprotein E-triglyceride rich lipoprotein particles: a major pathway at physiological particle concentrations. AB - We explored potential mechanisms of non-low-density lipoprotein (LDL) receptor mediated uptake of triglyceride-rich particles (TGRP) in the presence of apolipoprotein E (apo E). Human fibroblasts were incubated with model intermediate-density lipoprotein- (IDL-) sized TGRP (10-1000 microg of neutral lipid/mL) containing apo E. The extent of receptor-mediated uptake of TGRP was assessed with (a) an anti-apo E monoclonal antibody, which blocks receptor interaction; (b) incubation with heparin; (c) normal vs LDL receptor-negative fibroblasts; and (d) receptor-associated protein (RAP) to determine the potential contribution of LDL receptor-related protein (LRP). Cell surface heparan sulfate proteoglycan- (HSPG-) mediated uptake was examined with or without the addition of heparinase and heparitinase to cell incubation mixtures. At low particle concentrations (250 microg of neutral lipid/mL), most (>/=60%) particle uptake and internalization was via HSPG-mediated pathways. This HSPG pathway did not involve classical lipoprotein receptors, such as LRP or the LDL receptor. These data suggest that in peripheral tissues, such as the arterial wall, apo E may act in TGRP as a ligand for uptake not only via the LDL receptor and LRP pathways but also via HSPG pathways that are receptor-independent. Thus, at physiologic particle concentrations apo E-TGRP can be bound and internalized in certain cells by relatively low affinity but high capacity HSPG-mediated pathways. PMID- 9335534 TI - Photochemical and biochemical properties of chicken blue-sensitive cone visual pigment. AB - Through low-temperature spectroscopy and G-protein (transducin) activating experiments, we have investigated molecular properties of chicken blue, the cone visual pigment present in chicken blue-sensitive cones, and compared them with those of the other cone visual pigments, chicken green and chicken red (iodopsin), and rod visual pigment rhodopsin. Irradiation of chicken blue at -196 degrees C results in formation of a batho intermediate which then converts to BL, lumi, meta I, meta II, and meta III intermediates with the transition temperatures of -160, -110, -40, -20, and -10 degrees C. Batho intermediate exhibits an unique absorption spectrum having vibrational fine structure, suggesting that the chromophore of batho intermediate is in a C6-C7 conformation more restricted than those of chicken blue and its isopigment. As reflected by the difference in maxima of the original pigments, the absorption maxima of batho, BL, and lumi intermediates of chicken blue are located at wavelengths considerably shorter than those of the respective intermediates of chicken green, red and rhodopsin, but the maxima of meta I, meta II, and meta III are similar to those of the other visual pigments. These facts indicate that during the lumi-to meta I transition, retinal chromophore changes its original position relative to the amino acid residues which regulate the maxima of original pigments through electrostatic interactions. Using time-resolved low-temperature spectroscopy, the decay rates of meta II and meta III intermediates of chicken blue are estimated to be similar to those of chicken red and green, but considerably faster than those of rhodopsin. Efficiency in activating transducin by the irradiated chicken blue is greatly diminished as the time before its addition to the reaction mixture containing transducin and GTP increases, while that by irradiated rhodopsin is not. The time profile is almost identical with those observed in chicken red and green. Thus, the faster decay of enzymatically active state is common in cone visual pigments, independent of their spectral sensitivity. PMID- 9335535 TI - Disaccharide polyphosphates based upon adenophostin A activate hepatic D-myo inositol 1,4,5-trisphosphate receptors. AB - The glyconucleotides adenophostin A and B are the most potent known agonists at type 1 inositol trisphosphate [Ins(1,4,5)P3] receptors, although their stuctures differ markedly from that of Ins(1,4,5)P3. Equilibrium competition binding with [3H]Ins(1,4,5)P3 and unidirectional 45Ca2+ flux measurements were used to examine the effects of adenophostin A in hepatocytes, which express predominantly type 2 Ins(1,4,5)P3 receptors. Both Ins(1,4,5)P3 (Kd = 8.65 +/- 0.98 nM) and adenophostin A (Kd = 0.87 +/- 0.20 nM) bound to a single class of [3H]Ins(1,4,5)P3-binding site and each fully mobilized the same intracellular Ca2+ pool; although, adenophostin A (EC50 = 10.9 +/- 0.7 nM) was more potent than Ins(1,4,5)P3 (EC50 = 153 +/- 11 nM). Working on the assumption that it is the phosphorylated glucose component of the adenophostins that mimics the critical features of Ins(1,4,5)P3, we synthesized various phosphorylated disaccharide analogs containing this structure. The novel disaccharide-based analogs, sucrose 3,4,3'-trisphosphate [Sucr(3,4,3')P3], alpha,alpha'-trehalose 3,4,3',4' tetrakisphosphate [Trehal(3,4,3',4')P4], alpha,alpha'-trehalose 2,4,3', 4' tetrakisphosphate [Trehal(2,4,3',4')P4], and methyl 3-O-(alpha-d-glucopyranosyl) beta-d-ribofuranoside 2,3', 4'-trisphosphate [Rib(2,3',4')P3], were all able to mobilize the same intracellular Ca2+ pool as Ins(1,4,5)P3 and adenophostin A; although, none was as potent as adenophostin A. The rank order of potency of the analogs, adenophostin A > Ins(1,4,5)P3 approximately Rib(2,3',4')P3 > Trehal(2,4,3',4')P4 > Glc(2',3,4)P3 approximately Trehal(3,4,3',4')P4 > Sucr(3,4,3')P3, was the same in radioligand binding and functional assays of hepatic Ins(1,4,5)P3 receptors. Both Rib(2,3',4')P3, which was as potent as Ins(1,4,5)P3, and Trehal(2,4,3',4')P4 bound with significantly higher affinity ( approximately 27 and approximately 3-fold, respectively) than the only active carbohydrate agonist of Ins(1,4,5)P3 receptors previously examined [Glc(2',3,4)P3]. We conclude that phosphorylated disaccharides provide novel means of developing high-affinity ligands of Ins(1,4,5)P3 receptors. PMID- 9335536 TI - Solution secondary structure of a bacterially expressed peptide from the receptor binding domain of Pseudomonas aeruginosa pili strain PAK: A heteronuclear multidimensional NMR study. AB - The C-terminal receptor binding region of Pseudomonas aeruginosa pilin protein strain PAK (residues 128-144) has recently been the target for the design of a synthetic peptide vaccine effective against multiple strains of P. aeruginosa infection. We have successfully cloned and bacterially expressed a 15N-labeled PAK pilin peptide spanning residues 128-144 of the intact PAK pilin protein, PAK 128-144(Hs145), and have determined the solution secondary structure of this peptide using heteronuclear multidimensional NMR spectroscopy. The oxidized recombinant peptide exists as a major (trans) and minor (cis) species in solution, arising from isomerization around the Ile138-Pro139 peptide bond. The pattern of NOEs, temperature coefficients, and coupling constants observed for the trans isomer demonstrate the presence of a type I beta-turn and a type II beta-turn spanning Asp134-Glu-Gln-Phe137 and Pro139-Lys-Gly-Cys142, respectively. This is in agreement with the NMR solution structure of the trans isomer of a synthetic PAK 128-144 peptide which showed a type I and a type II beta-turn in these same regions of the sequence [McInnes, C., Sonnichsen, F. D., Kay, C. M., Hodges, R. S., and Sykes, B. D. (1993) Biochemistry 32, 13432-13440; Campbell, A. P., McInnes, C., Hodges, R. S., and Sykes, B. D. (1995) Biochemistry 34, 16255 16268]. The pattern of NOEs, temperature coefficients, and coupling constants observed for the cis isomer also demonstrate a type II beta-turn spanning Pro139 Lys-Gly-Cys142, but suggest a second beta-turn spanning Asp132-Gln-Asp-Glu135. Thus, the cis isomer may also possess a double-turn motif (like the trans isomer), but with different spacing between the turns and a different placement of the first turn in the sequence. The discovery of a double-turn motif in the trans (and cis) recombinant PAK pilin peptide is an extremely important result since the double turn has been implicated as a structural requirement for the recognition of both receptor and antibody. These results pave the way for future isotope-edited NMR studies of the labeled recombinant PAK pilin peptide bound to antibody and receptor, studies integral to the design of an effective synthetic peptide vaccine. PMID- 9335537 TI - Probing the roles of active site residues in phosphatidylinositol-specific phospholipase C from Bacillus cereus by site-directed mutagenesis. AB - The role of amino acid residues located in the active site pocket of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus[Heinz, D. W., Ryan, M., Bullock, T., & Griffith, O. H. (1995) EMBO J. 14, 3855-3863] was investigated by site-directed mutagenesis, kinetics, and crystal structure analysis. Twelve residues involved in catalysis and substrate binding (His32, Arg69, His82, Gly83, Lys115, Glu117, Arg163, Trp178, Asp180, Asp198, Tyr200, and Asp274) were individually replaced by 1-3 other amino acids, resulting in a total number of 21 mutants. Replacements in the mutants H32A, H32L, R69A, R69E, R69K, H82A, H82L, E117K, R163I, D198A, D198E, D198S, Y200S, and D274S caused essentially complete inactivation of the enzyme. The remaining mutants (G83S, K115E, R163K, W178Y, D180S, Y200F, and D274N) exhibited reduced activities up to 57% when compared with wild-type PI-PLC. Crystal structures determined at a resolution ranging from 2.0 to 2.7 A for six mutants (H32A, H32L, R163K, D198E, D274N, and D274S) showed that significant changes were confined to the site of the respective mutation without perturbation of the rest of the structure. Only in mutant D198E do the side chains of two neighboring arginine residues move across the inositol binding pocket toward the newly introduced glutamic acid. An analysis of these structure-function relationships provides new insight into the catalytic mechanism, and suggests a molecular explanation of some of the substrate stereospecificity and inhibitor binding data available for this enzyme. PMID- 9335538 TI - Failure of a two-state model to describe the influence of phospho(enol)pyruvate on phosphofructokinase from Escherichia coli. AB - A linked-function analysis is presented of the influence of the inhibitor phospho(enol)pyruvate (PEP) on the binding of fructose 6-phosphate (Fru-6-P) and MgATP to phosphofructokinase (PFK) from Escherichia coli. The results of this analysis indicate that the previously described inhibition of Fru-6-P binding by MgATP [Johnson, J. L., & Reinhart, G. D. (1992) Biochemistry 31, 11510-11518] is almost completely independent of the inhibition by PEP. Moreover, with or without the presence of MgATP, the inhibition by PEP does not conform to the behavior expected if PEP and Fru-6-P bind exclusively to different enzyme forms since the formation of a ternary complex with both PEP and Fru-6-P bound is clearly evident at high concentrations of Fru-6-P and PEP. van't Hoff analyses of the coupling interactions between PEP and Fru-6-P in the presence and absence of MgATP indicate that these couplings are driven by enthalpy. However, the influence that PEP has on MgATP binding is small and changes from being activating to being inhibitory at temperatures above 40 degrees C, revealing the importance of a compensating entropy component to the coupling interactions. The four functionally defined enzyme forms that contribute to the coupling between Fru-6-P and PEP were evaluated structurally using the fluorescence properties of the single intrinsic tryptophan as a probe. The steady-state and dynamic fluorescence emission and polarization properties of the tryptophan, as well as its susceptibility to I- quenching, indicate that the flexibility of PFK in the vicinity of the tryptophan is perturbed by the binding of ligands. The properties of free PFK do not lie between those established by the binding of Fru-6-P and PEP individually, indicating that it is structurally distinct. The properties of the ternary complex lie between those of the singly-ligated forms. Though an equilibrium mixture of two conformations of ternary complex cannot therefore be formally ruled out, no evidence obtained to date requires the presumption of this mechanistic complication. PMID- 9335539 TI - Calcium/calmodulin-dependent protein kinase kinase: identification of regulatory domains. AB - We recently cloned a calmodulin-dependent protein kinase kinase (CaM-KK) which phosphorylates and activates CaM-KI and CaM-KIV [Tokumitsu, H., Enslen, H., and Soderling, T. R. (1995) J. Biol. Chem. 270, 19320-19324]. In the present study, we have identified its regulatory CaM-binding and autoinhibitory domains (CBD and AID, respectively) using a series of COOH-terminal truncations and site-directed mutants expressed in COS-7 cells. Truncation mutant CaM-KK1-463 activated CaM-KIV and bound CaM similar to wild-type enzyme (CaM-KK1-505); CaM-KK1-448 did not bind CaM and was largely inactive; and CaM-KK1-434 also did not bind CaM but activated a CaM-independent mutant of CaM-KIV in the absence of Ca2+/CaM. Substitution of triple negative charges (Asp) at positions 455RKR, 448ILV, or 443SWT blocked CaM binding and suppressed by 70-90% CaM-KK activities. Mutants 438VKL and 435KNS to DDD exhibited partial Ca2+/CaM-independent activities. These results identify overlapping AID and CBD between residues 430 and 460 in CaM-KK, similar to other CaM-Ks. Consistent with this assignment, the synthetic peptide corresponding to residues 438-463 bound CaM in a Ca2+-dependent manner with a Kd in the low nanomolar range. Furthermore, phosphorylation by cAMP-kinase of Ser458 at the COOH-terminus of the CBD in CaM-KK, which suppresses subsequent CaM binding [Wayman, G., Tokumitsu, H., and Soderling, T. R. (1997) J. Biol. Chem. 272, 16073 16076], was blocked by prior binding of Ca2+/CaM to CaM-KK. PMID- 9335540 TI - Role of guanine nucleotides in the vinblastine-induced self-association of tubulin: effects of guanosine alpha,beta-methylenetriphosphate and guanosine alpha,beta-methylenediphosphate. AB - It is now well established that guanine nucleotides are allosteric effectors of the vinca alkaloid-induced self-association of tubulin. GDP enhances self association for vinblastine-, vincristine- and vinorelbine-induced spiral assembly relative to GTP by 0.90 +/- 0.17 kcal/mol [Lobert et al. (1996) Biochemistry 35, 6806-6814]. Since chemical modifications of the vinca alkaloid structure are known to modulate the overall affinity of drug binding, it is very likely that, by Wyman linkage, chemical modifications of guanine nucleotide allosteric effectors also modulate drug binding. Here we compare the effects of the GTP and GDP alpha,beta-methylene analogues GMPCPP and GMPCP on vinblastine induced tubulin association in 10 and 100 mM piperazine-N,N'-bis(2-ethanesulfonic acid) (Pipes), 1 mM MgSO4, and 2 mM [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA), pH 6. 9, at different temperatures. We found that GMPCPP perfectly mimics GTP in its effect on spiral assembly under all ionic strength and temperature conditions. However, GMPCP in 10 mM Pipes behaves not as a GDP analogue, but as a GTP analogue. In 100 mM Pipes, GMPCP has characteristics that are intermediate between GDP and GTP. These data suggest that the alpha,beta methylene group in GMPCP and GMPCPP is sufficient to produce a GTP-like effect on vinblastine-induced tubulin self-assembly. This is consistent with previous observations that GMPCP-tubulin will assemble into microtubules in a 2 M glycerol and 100 mM Pipes buffer [Vulevic & Correia (1997) Biophys. J. 72, 1357-1375]. Our results demonstrate that an alpha,beta methylene modification of the guanine nucleotide phosphate moiety can induce a salt-dependent conformational change in the tubulin heterodimer that favors the GTP-tubulin structure. This has important implications for understanding allosteric interactions that occur in the binding of guanine nucleotides to tubulin. PMID- 9335541 TI - Nonidentity of the alpha-neurotoxin binding sites on the nicotinic acetylcholine receptor revealed by modification in alpha-neurotoxin and receptor structures. AB - alpha-Neurotoxins constitute a large family of polypeptides that bind with high affinity to the nicotinic acetylcholine receptor (nAChR). Using a recombinant DNA derived alpha-neurotoxin (Naja mossambica mossambica, NmmI) and mouse muscle nAChR expressed transiently on the surface of HEK 293 cells, we have delineated residues involved in the binding interaction on both the alpha-neurotoxin and the receptor interface. Several of the studied NmmI mutations, including two residues conserved throughout the alpha-neurotoxin family (K27 and R33), resulted in substantial decreases in the binding affinity. We have also examined 23 mutations located on the receptor alpha subunit and have identified 4 positions that appear to be important to NmmI recognition. These determinants represent a conserved aromatic residue (Y190), two positions where neuronal and muscle receptors differ (V188 and P197), and a negatively charged residue (D200). Unlike many of the nAChR agonists and antagonists which bind to the alphadelta and alphagamma binding sites on the receptor with different affinities, the wild-type NmmI-wild type nAChR interaction showed a single affinity. However, by mutating critical toxin or receptor residues, we were able to produce site-selectivity between the alphagamma and alphadelta interfaces. These results suggest a nonequivalence in the binding interaction at the two sites, sensitive to discrete structural changes at key contact points on either the toxin or the receptor protein, and underscore the importance of delta and gamma receptor subunits in governing binding affinity. PMID- 9335542 TI - Perturbations of functional interactions with myosin induce long-range allosteric and cooperative structural changes in actin. AB - The role of the rotational dynamics of actin filaments in their interaction with myosin was studied by comparing the effect of myosin subfragment 1 (S1) with two other structural perturbations, which have substantial inhibitory effects on activation of myosin ATPase and in vitro motility of F-actin: (1) binding of the antibody fragment Fab(1-7) against the first seven N-terminal residues and (2) copolymerization with monomers treated with the zero-length cross-linker 1-ethyl 3-[3-(dimethylamino)propyl]carbodiimide (EDC), referred to as EDC-actin. The rotational motion of actin was measured by time-resolved phosphorescence anisotropy (TPA) of erythrosin iodoacetamide (ErIA) attached to Cys 374 on actin. The binding of S1 in a rigor complex (no nucleotide) induced intramonomer (allosteric) and intermonomer (cooperative) structural changes that increased the residual anisotropy of labeled F-actin, indicating a conformational change in the region of the C terminus. Similar allosteric and cooperative changes were induced by binding of Fab(1-7) and by copolymerization of the ErIA-labeled actin monomers with EDC-actin. This suggests that the functional perturbations transform actin to a form resembling the rigor actomyosin complex. The correlation of the perturbation-induced changes in TPA of actin with the functional effects suggests that the actomyosin interaction can be inhibited by stabilization of actin in one of its structural intermediates. PMID- 9335543 TI - Calmodulin-binding autoinhibitory domain controls "pH-sensing" in the Na+/H+ exchanger NHE1 through sequence-specific interaction. AB - The calmodulin (CaM)-binding domain reduces the affinity of the Na+/H+ exchanger NHE1 for intracellular H+ by exerting an autoinhibitory function in quiescent cells. We replaced this domain (aa 637-656) with homologous segments from other NHE isoforms (NHE2 and 4) or functionally similar regions from other sources (Na+/Ca2+ exchanger, CaM-dependent protein kinase II, plasma membrane Ca2+-pump, or CaM-binding peptide Trp3). The NHE-1-, NHE2-, and NHE4-segments bound CaM with Kds of 16, 130, and 27 nM, respectively. These chimeric molecules were expressed in the exchanger-deficient cell PS120. NHE1 with incorporated NHE2-segment was activated in response to Ca2+-mobilizing agents ionomycin and thrombin resulting in an alkaline shift of the intracellular pH (pHi)-dependence of 22Na+ uptake, as was the case with the intact rat NHE2. In contrast, incorporation of the NHE4 segment or other CaM-binding segments induced a constitutive alkaline shift of pHi-dependence with concomitant abolishment of Ca2+-dependent activation, indicating that these segments could not function as an autoinhibitory domain in NHE1. Detailed analyses revealed that Leu639, Lys651 and Tyr652, conserved in the NHE1- and NHE2-segments, but not in the NHE4-segment, are important for the autoinhibition. Furthermore, 125I-labeled CaM-binding peptide from NHE1 was efficiently crosslinked to the NHE1 protein, suggesting that the inhibitory domain physically interacts with part(s) of the molecule. Together, these findings support the notion that the reduction of H+ affinity in Na+/H+ exchange occurs through a mechanism involving a highly sequence-specific interaction of the inhibitory domain with its putative acceptor in NHE1. PMID- 9335544 TI - Interaction of the delta-endotoxin CytA from Bacillus thuringiensis var. israelensis with lipid membranes. AB - We investigated the binding of CytA, a cytolytic delta-endotoxin from Bacillus thuringiensis var. israelensis, to small unilamellar lipid vesicles (SUV) and the accompanying changes in the overall CytA conformation. From the titration of tryptophan fluorescence with SUV, we determined the apparent association constants of 3500 M-1 and 11000 M-1 for the protoxin CytA27 and the proteolytically activated toxin CytA24, respectively. Inclusion of a negatively charged lipid or a positively charged lipid analog in the membrane did not affect the binding parameters, which suggests that membrane binding is not driven by electrostatic interactions. A decrease in the intensity of the CytA tryptophan fluorescence upon interaction with lipids and the absence of a blue shift in remaining fluorescence indicate that the tryptophan-containing regions of the protein do not significantly penetrate into the hydrophobic core of the lipid bilayer. This finding was corroborated by the lack of additional quenching by brominated or spin-labeled lipids, irrespective of the location of the quenching moiety in the depth of the bilayer. However, the interaction with lipids decreases quenching with the soluble quenchers acrylamide and KI, and the remaining fluorescence is blue-shifted. The observed decrease in fluorescence anisotropy upon membrane binding is not consistent with simple immobilization of CytA on the surface of SUV. We showed by FTIR spectroscopy and differential scanning calorimetry (DSC) that binding to the membrane causes a significant loosening of the protein structure. This is consistent with the fluorescence quenching and anisotropy data. Our experiments provide evidence against CytA's substantially penetrating the lipid bilayer and creating well-defined proteinaceous channels. PMID- 9335545 TI - Influence of the angle subtended by the positively charged helix face on the membrane activity of amphipathic, antibacterial peptides. AB - To investigate the influence of the angle subtended by the positively charged helix face on membrane activity, six amphipathic alpha-helical peptides with angles between 80 degrees and 180 degrees, but with retained hydrophobicity, hydrophobic moment, and positive overall charge, were designed starting from the sequence of the antibacterial peptide magainin 2. CD investigations revealed that all analogs are in an alpha-helical conformation in vesicle suspension. The ability of the peptides to induce dye release from negatively charged phosphatidylglycerol (PG) vesicles decreased with increasing angle. However, peptides with a large angle of positively charged residues (140-180 degrees) exhibited a considerably higher permeabilizing activity at zwitterionic phosphatidylcholine (PC) and mixed PC/PG (3:1) vesicles than analogs with a small angle (80-120 degrees). In addition, analogs with large angles were more active in antibacterial and hemolytic assays. The antibacterial specificity of these analogs was decreased. Binding investigations showed that peptide binding is favored by a large angle and a high content of negatively charged phospholipid. In contrast, a small angle and a low negative membrane charge enhanced the membrane-permeabilizing efficiency of the bound peptide fraction. All analogs stabilized the bilayer phase of phosphatidylethanolamine over the inverted hexagonal phase. Therefore, a class L mechanism of permeabilization can be excluded. Furthermore, the analogs do not act by the induction of positive curvature strain or by a "carpet-like" mechanism. Our results are in accordance with a pore mechanism: The membrane-permeabilizing efficiency of analogs with enhanced angle of positively charged residues is reduced due to electrostatic repulsion between adjacent helices within the pore, thus resulting in a decreased pore-forming probability and/or pore destabilization. PMID- 9335546 TI - Irreversible activation of the gonadotropin-releasing hormone receptor by photoaffinity cross-linking: localization of attachment site to Cys residue in N terminal segment. AB - Photoaffinity cross-linking with [azidobenzoyl-d-Lys6]GnRH leads to irreversible activation of the gonadotropin-releasing hormone (GnRH) receptor. In order to localize the cross-linking site, the disulfide bridge structure was initially probed by mutagenesis. A consistent pattern of changes in the ability of GnRH to stimulate signal transduction after Ser substitutions of extracellularly located Cys residues indicated that Cys14 in the N-terminal domain is connected to Cys200 in the second extracellular loop, while Cys196 in this loop is connected to the highly conserved Cys114 at the extracellular end of transmembrane helix 3. Protein chemical analysis of radioactive fragments of cross-linked GnRH receptor following deglycosylation and enzymatic digest with endoproteinase Glu-C and trypsin before and after introduction or elimination of potential protease cleavage sites indicated that 125I[azidobenzoyl-d-Lys6]GnRH cross-links to a segment comprising residues 12-18 of the N-terminal domain. The existence of the Cys114-Cys196 bridge was directly confirmed as a labeled fragment, including that Cys114 was resolvable only under reducing conditions. The observation that the cross-linked N-terminal enzymatic fragments had identical apparent size under non reducing conditions shows that the cross-linking reaction disconnected the disulfide bridge between Cys14 and Cys200 and indicates that Cys14 is probably the residue involved in cross-linking of the ligand. It is concluded that covalent tethering of GnRH through a photoreactive side chain located at position 6 in the middle of the peptide leads to continued activation of the receptor presumably through covalent binding to Cys14 in the N-terminal domain of the receptor. PMID- 9335547 TI - Self-phosphorylation of epidermal growth factor receptor is an intermolecular reaction. AB - The binding of epidermal growth factor (EGF) to epidermal growth factor receptor (EGF receptor) results in the dimerization and self-phosphorylation of the receptor. Both of these responses were followed as a function of time and the concentration of EGF receptor. Dimerization of EGF receptor was monitored by immunoblotting the protein after it had been cross-linked with glutaraldehyde. The capacity for self-phosphorylation was followed by measuring the relative level of incorporation of [32P]phosphate into EGF receptor on autoradiograms of the same immunoblots used for the assay of its dimerization. When these two properties were followed as a function of time, it was found that dimerization preceded the appearance of the capacity for self-phosphorylation. Both dimeric and monomeric forms of EGF receptor were self-phosphorylated in the presence of EGF, but the dimeric form was phosphorylated preferentially to the monomeric form. When the dimerization and the capacity for self-phosphorylation were followed as a function of the concentration of dimeric EGF receptor, it was observed that the self-phosphorylation of dimeric EGF receptor increased as the concentration of dimeric EGF receptor increased. An equation including terms representing both intramolecular and intermolecular rates of self-phosphorylation was fit to the plots of self-phosphorylation as a function of concentration of EGF receptor. These fits demonstrate that intramolecular self-phosphorylation within dimers of EGF receptor is insignificant and that self-phosphorylation is an intermolecular process between dimers of EGF receptor. PMID- 9335548 TI - Role of conserved arginine and glutamate residues on the cytosolic surface of glucose transporters for transporter function. AB - The role of conserved arginine and glutamic acid residues at the cytoplasmic surface of the GLUT4 for transporter function was investigated by site-directed mutagenesis and expression of the constructs in COS-7 cells. Reconstituted glucose transport activity, cytochalasin B binding, and photolabeling with the exofacial label 2-N4-(1-azi-2,2,2-trifluoroethyl)benzoyl-1, 3-bis(d-mannosyloxy) 2-propylamine (ATB-BMPA) was assayed in membranes from transfected cells and corrected for immunoreactivity of expressed transporters. Exchange of Arg 92 (R92L amino acid residues are numbered according to the corresponding residues in the GLUT1) or Arg 333/334 (RR333/4LA) reduced or suppressed transport activity with no or very little effect on photolabeling with ATB-BMPA and cytochalasin B binding. It is suggested that the lack of these residues selectively disturbes the substrate-induced conformational change of the carrier during transport. Exchange of Glu 146 (E146D) or Arg 153 (R153L) markedly reduced transport activity, ATB-BMPA photolabeling, and cytochalasin B binding. Transport activity and ATB-BMPA labeling were abolished in the mutants E329Q, E393D, and R400L, whereas binding of cytochalasin B was normal. Thus, exchange of Glu 329, Glu 393, and Arg 400 appears to arrest the transporter in an inward facing conformation. It is concluded that the conserved arginine and glutamate residues at the cytoplasmic surface of the glucose transporter GLUT4 are essential for its appropriate conformation, and that it is the interaction of charged residues which mediates the oscillation between outward and inward facing states. PMID- 9335549 TI - Comparison of the kinetic effects of phospholamban phosphorylation and anti phospholamban monoclonal antibody on the calcium pump in purified cardiac sarcoplasmic reticulum membranes. AB - Protein kinase A- (PKA-) catalyzed phosphorylation of phospholamban (PLN), the protein regulator of the cardiac Ca pump, mediates abbreviation of systole in response to beta-adrenergic agonists. Investigators previously, however, have been unsuccessful in demonstrating an effect of PLN phosphorylation or anti-PLN monoclonal antibody (mAb), which is considered to mimic phosphorylation's well known effect on Km(Ca), on microsomal Ca uptake at the (high) Ca2+ concentrations found intracellularly at peak systole. We therefore compared the effects of the catalytic subunit of PKA and anti-PLN mAb on the kinetics of Ca uptake in sucrose gradient-purified cardiac microsomes. Both treatments produced a 33-44% increase in Vmax(Ca) at 25 and 37 degrees C, and an 11-31% decrease in Km(Ca) with comparable changes in Ca2+-ATPase activity. An acceleration of E2P decomposition upon PLN phosphorylation may contribute to the increased Vmax(Ca) of Ca uptake at 25 degrees C but not at 37 degrees C, based on measurement of the kinetics of E2P decomposition and steady-state E2P formation from Pi at different temperatures. Our data document almost identical increases in Vmax(Ca) of microsomal Ca uptake with PLN phosphorylation or addition of anti-PLN mAb and hence provide insight into the kinetic mechanism of PLN's regulation of the cardiac sarcoplasmic reticulum Ca pump protein. PMID- 9335550 TI - Determination of singlet oxygen-specific versus radical-mediated lipid peroxidation in photosensitized oxidation of lipid bilayers: effect of beta carotene and alpha-tocopherol. AB - Photosensitized oxidation reactions damage tissue by catalyzing the formation of oxyradicals and singlet oxygen. beta-Carotene is hypothesized to exert photoprotective effects by quenching singlet oxygen formed by Type II reactions and by scavenging free radicals formed by Type I reactions. beta-Carotene antioxidant mechanisms were studied in a phospholipid membrane model of photooxidation with a new isotope dilution gas chromatography-mass spectrometry (GC-MS) assay that quantitatively distinguishes singlet oxygen-mediated and radical-mediated lipid peroxidation. This assay measures 9- and 10 hydroxylinoleate methyl esters and was used to generate photooxidation profiles for the photosensitizers methylene blue, Rose Bengal, and tetraphenylporphine. These profiles indicate a shift from Type II to Type I photooxidation mechanisms in later stages of photooxidation. beta-Carotene (0.45 mol %) inhibited singlet oxygen-mediated lipid peroxidation at early stages of methylene blue-sensitized photooxidation. Production of radical-mediated products increased faster than singlet oxygen-mediated products at later stages. beta-Carotene-5,8-endoperoxide, a specific marker for singlet oxygen oxidation of beta-carotene in solution, was unstable under the incubation conditions and was not detected in this system. alpha-Tocopherol (0.45 mol %) was ineffective in inhibiting photosensitized lipid peroxidation, whereas 4.5 mol % alpha-tocopherol inhibited almost all radical mediated lipid peroxidation as well as early-stage singlet oxygen-mediated lipid peroxidation. Cumene hydroperoxide stimulated radical-mediated lipid peroxidation, indicating that accumulation of hydroperoxides from Type II photooxidation may enhance Type I reactions. These data suggest that singlet oxygen quenching, rather than radical scavenging reactions, accounts for the photoprotective actions of beta-carotene. PMID- 9335551 TI - Cytoplasmic domains mediate the ligand-induced affinity shift of guanylyl cyclase C. AB - Guanylyl cyclase C (GCC), the receptor for the Escherichia coli heat-stable enterotoxin (ST), exhibits multiple binding affinities, including high (RH) and low (RL) affinity sites and a ligand-induced conversion of low-affinity sites from a higher (RL1) to a lower (RL2) affinity state. Occupancy of the lowest affinity state of low-affinity sites is coupled to ligand-induced catalytic activation. In the present studies, ligand binding and catalytic activation properties of a series of intracellular deletion mutants of GCC were examined to identify the structural domains underlying expression of high- and low-affinity sites and the ligand-induced shift in low-affinity sites. These studies demonstrated that the cytoplasmic domains of GCC are not required, but extracellular and transmembrane domains are sufficient, for expression of high affinity binding sites. In addition, the cytoplasmic juxtamembrane and kinase homology domains are required for expression of the ligand-induced affinity shift in low-affinity sites. Of significance, this shift in affinity was insensitive to adenine nucleotides, in contrast to other members of the receptor guanylyl cyclase family, such as guanylyl cyclase A (GCA). Also, the juxtamembrane and kinase homology domains are critical for coupling ST-receptor binding and guanylyl cyclase catalytic activation. Indeed, deletion of those domains from GCC results in a constitutively inhibited enzyme, suggesting that they function as positive effectors of ligand activation, in contrast to GCA and GCB, in which the kinase homology domain represses basal catalytic activity. These data suggest that the mechanisms regulating different members of the receptor guanylyl cyclase family are overlapping but not identical. PMID- 9335553 TI - Protein kinase C modulation of insulin receptor substrate-1 tyrosine phosphorylation requires serine 612. AB - Activation of the endogenous protein kinase Cs in human kidney fibroblast (293) cells was found in the present study to inhibit the subsequent ability of insulin to stimulate the tyrosine phosphorylation of an expressed insulin receptor substrate-1. This inhibition was also observed in an in vitro phosphorylation reaction if the insulin receptor and its substrate were both isolated from cells in which the protein kinase C had been activated. To test whether serine phosphorylation of the insulin receptor substrate-1 was contributing to this process, serine 612 of this molecule was changed to an alanine. The insulin stimulated tyrosine phosphorylation and the associated phosphatidylinositol 3 kinase activity of the expressed mutant were found to be comparable to those of the expressed wild-type substrate. However, unlike the wild-type protein, activation of protein kinase C did not inhibit the insulin-stimulated tyrosine phosphorylation of the S612A mutant nor its subsequent association with phosphatidylinositol 3-kinase. Tryptic peptide mapping of in vivo labeled IRS-1 and the S612A mutant revealed that PMA stimulates the phosphorylation of a peptide from wild-type IRS-1 that is absent from the tryptic peptide maps of the S612A mutant. Moreover, a synthetic peptide containing this phosphoserine and its nearby tyrosine was found to be phosphorylated by the insulin receptor to a much lower extent than the same peptide without the phosphoserine. Activation of protein kinase C was found to stimulate by 10-fold the ability of a cytosolic kinase to phosphorylate this synthetic peptide as well as the intact insulin receptor substrate-1. Finally, cytosolic extracts from the livers of ob/ob mice showed an 8-fold increase in a kinase activity capable of phosphorylating this synthetic peptide, compared to extracts of livers from lean litter mates. These results indicate that activation of protein kinase C stimulates a kinase which can phosphorylate insulin receptor substrate-1 at serine 612, resulting in an inhibition of insulin signaling in the cell, posing a potential mechanism for insulin resistance in some models of obesity. PMID- 9335552 TI - Human 5-HT1 receptor subtypes exhibit distinct G protein coupling behaviors in membranes from Sf9 cells. AB - The G protein coupling behavior of four human 5-hydroxytryptamine receptor subtypes (5-HT1A, 5-HT1B, 5-HT1D, and 5-HT1E) has been studied in membranes from Sf9 cells expressing the individual receptors. The 5-HT1A and 5-HT1B receptors exhibited both high- and low-affinity states for agonist, with the majority of the receptors in a low-affinity state. Addition of purified G protein subunits to membranes expressing either 5-HT1A or 5-HT1B receptors shifted the majority of the receptors to a high-affinity state in the absence, but not in the presence, of guanine nucleotides. The alphai1, alphai2, alphai3, and alphao subunits were able to shift the receptors to a high-affinity state with either betagammabrain or betagammaretina while alphat subunits were inactive regardless of which betagamma preparation was used. A significantly higher affinity for agonist was observed with both receptors in the presence of alphai3 subunits compared with either alphai2 or alphao subunits, while a significantly lower concentration of alpha subunits was required for a maximal affinity shift of 5-HT1A receptors compared with 5-HT1B receptors (EC50 values of 6.4 and 12. 0 nM, respectively). The 5-HT1D and 5-HT1E receptors exhibited only a single affinity state for agonist. Addition of purified G protein subunits to membranes containing 5-HT1D receptors caused a small increase in affinity for agonist that was only partially reversed by guanine nucleotides while the addition of purified G protein subunits to membranes containing 5-HT1E receptors had no affect on agonist binding. Thus when expressed in an identical membrane environment these four closely related 5 HT1 receptor subtypes exhibit different G protein coupling behaviors. PMID- 9335554 TI - Mechanism of action of the unusually potent microtubule inhibitor cryptophycin 1. AB - Cryptophycin 1 is a remarkably potent antiproliferative compound that shows excellent antitumor activity against mammary, colon, and pancreatic adenocarcinomas in mouse xenographs. At picomolar concentrations, cryptophycin 1 blocks cells in the G2/M phase of the cell cycle by an apparent action on microtubules. The compound binds to tubulin, inhibits microtubule polymerization, and depolymerizes preformed microtubules in vitro. Its exceptionally powerful antitumor activity (many-fold greater than paclitaxel or the vinca alkaloids) raises important questions about its mechanism of action. By quantitative video microscopy, we examined the effects of cryptophycin 1 on the dynamics of individual microtubules assembled to steady state from bovine brain tubulin. At low nanomolar concentrations, in the absence of net microtubule depolymerization, cryptophycin 1 potently stabilized microtubule dynamics. It reduced the rate and extent of microtubule shortening and growing and increased the frequency of rescue. The results suggest that cryptophycin 1 exerts its antiproliferative and antimitotic activity by binding reversibly and with high affinity to the ends of microtubules, perhaps in the form of a tubulin-cryptophycin 1 complex, resulting in the most potent suppression of microtubule dynamics yet described. PMID- 9335555 TI - Mechanism of energy coupling in the FOF1-ATP synthase: the uncoupling mutation, gammaM23K, disrupts the use of binding energy to drive catalysis. AB - The Escherichia coli FOF1 ATP synthase uncoupling mutation, gammaM23K, was found to increase the energy of interaction between gamma and beta subunits which caused inefficient transmission of coupling information between transport and catalysis [Al-Shawi, M. K. , Ketchum, C. J., and Nakamoto, R. K. (1997) J. Biol. Chem. 272, 2300-2306]. We hypothesized that the gammaM23K mutation, because of its effect on coupling, should alter the fundamental reactions steps that are normally modulated by DeltamuH+ via the coupling mechanism. In this paper, we address this issue by studying the thermodynamics of individual catalytic steps through the use of energy profiles to gain information regarding enzyme mechanism and the effects of the mutation. Compared to wild-type enzyme, the gammaM23K F1 had significant differences of two partial reactions: the rate constant for Pi release was 49-fold faster and the rate constant for ATP release was 8.4-fold faster than wild-type. These rate constants were considered together with characteristics of a group of F1 ATPase mutant enzymes and were analyzed quantitatively by linear free energy relationships [Al-Shawi, M. K., Parsonage, D., and Senior, A. E., (1990) J. Biol. Chem. 265, 4402-4410]. We found that the gammaM23K mutation prevents the proper utilization of binding energy to drive catalysis and blocks the enzyme in a Pi release mode. This finding is consistent with the use of energy from DeltamuH+ for increasing the affinity for Pi so that the substrate binds in a catalytically competent manner for synthesis of ATP. These results support the notion that the communication of coupling information is transmitted through the gamma-beta interface near gammaMet23 and beta380DELSEED386 segment. PMID- 9335556 TI - The Escherichia coli FOF1 gammaM23K uncoupling mutant has a higher K0.5 for Pi. Transition state analysis of this mutant and others reveals that synthesis and hydrolysis utilize the same kinetic pathway. AB - The Escherichia coli FOF1 ATP synthase uncoupling mutation, gammaM23K, was found to increase the energy of interaction between gamma and beta subunits, prevent the proper utilization of binding energy to drive catalysis, and block the enzyme in a Pi release mode. In this paper, the effects of this mutation on substrate binding in cooperative ATP synthesis are assessed. Activation of ATP synthesis by ADP and Pi was determined for the gammaM23K FOF1. The K0.5 for ADP was not affected, but K0.5 for Pi was approximately 7-fold higher even though the apparent Vmax was close to the wild-type level. Wild-type enzyme had a turnover number of 82 s-1 at pH 7.5 and 30 degrees C. During oxidative phosphorylation, the apparent dissociation constant (KI) for ATP was not affected and was 5-6 mM for both wild-type and gammaM23K enzymes. Thus, the apparent binding affinity for ATP in the presence of DeltamuH+ was lowered by 7 orders of magnitude from the affinity measured at the high-affinity catalytic site. Arrhenius analysis of ATP synthesis for the gammaM23K FOF1 revealed that, like those of ATP hydrolysis, the transition state DeltaH was much more positive and TDeltaS was much less negative, adding up to little change in DeltaG. These results suggested that ATP synthesis is inefficient because of an extra bond between gamma and beta subunits which must be broken to achieve the transition state. Analysis of the transition state structures using isokinetic plots demonstrate that ATP hydrolysis and synthesis utilize the same kinetic pathway. Incorporating this information into a model for rotational catalysis suggests that at saturating substrate concentrations, the rate-limiting step for hydrolysis and synthesis is the rotational power stroke where each of the beta subunits changes conformation and affinity for nucleotide. PMID- 9335557 TI - Solvent accessibility of the phycocyanobilin chromophore in the alpha subunit of C-phycocyanin: implications for a molecular mechanism for inertial protein-matrix solvation dynamics. AB - The solvent environment of the phycocyanobilin chromophore bound by the alpha subunit of C-phycocyanin was probed in buffered binary solvent systems consisting of water and methanol, acetonitrile, or acetone. The focus of the work was on determining whether the inertial phase of the solvent response observed previously in the alpha subunit from femtosecond transient hole-burning spectroscopy [Riter et al. (1996) J. Phys. Chem. 100, 14198-14205] involves solvent dipoles in the bulk. Continuous absorption and fluorescence spectra at room temperature show that addition of the nonaqueous solvent results in a change in the tertiary structure of the protein so that the phycocyanobilin chromophore is unclamped and allowed to assume a cyclic conformation. At low concentrations of nonaqueous solvent, we observe a conformational equilibrium characterized by a cooperative binding of nonaqueous solvent. The phycocyanobilin chromophore exhibits a nonshifted absorption and fluorescence spectrum characteristic of its native, extended conformation in the state with bound water molecules. In the state with bound solvent molecules, the phycocyanobilin chromophore exhibits an absorption spectrum that reports a cyclic configuration, and its fluorescence is essentially quenched. The absorption and fluorescence spectra exhibit a solvatochromic response in this state, indicating that the chromophore is now exposed to the bulk solvent. Far-UV circular dichroism spectra evidence an abrupt loss of 10% of the alpha-helical character in the nonaqueous solvent concentration regime that results in exposure of the chromophore to the bulk. These results show that the ultrafast solvation response previously detected in the alpha subunit in aqueous media from femtosecond transient hole-burning spectroscopy arises solely from protein-matrix solvation dynamics. PMID- 9335558 TI - Changes in the midpoint potentials of the nitrogenase metal centers as a result of iron protein-molybdenum-iron protein complex formation. AB - All nitrogenase-catalyzed substrate reduction reactions require the transient association between the iron (Fe) protein component and the molybdenum-iron (MoFe) protein component with concomitant intercomponent electron transfer and MgATP hydrolysis. Understanding the effects of Fe protein-MoFe protein complex formation on the properties of the nitrogenase metal centers is thus essential to understanding the electron transfer reactions. This work presents evidence for significant shifts in midpoint potentials for two of the three nitrogenase metal centers as a result of Fe protein binding to the MoFe protein. The midpoint potentials for the three nitrogenase metal centers, namely the [4Fe-4S] cluster of the Fe protein, and the [8Fe-7S] (or P-) clusters and FeMo cofactors (or M centers) of the MoFe protein, were determined within a nondissociating nitrogenase complex prepared with a site-specifically altered Fe protein (Leu at position 127 deleted, L127Delta). The midpoint potential for each metal center was determined by mediated redox titrations, with the redox state of each center being monitored by parallel and perpendicular mode EPR spectroscopy. The midpoint potential of the Fe protein [4Fe-4S]2+/1+ cluster couple was observed to change by -200 mV from -420 mV in the uncomplexed L127Delta Fe protein to -620 mV in the L127Delta Fe protein-MoFe protein complex. The midpoint potential of the two electron oxidized couple of the P-clusters (P2+/N) of the MoFe protein was observed to shift by -80 mV upon protein-protein complex formation. No significant change in the midpoint potential of an oxidized state of FeMoco (Mox/N) was observed upon complex formation. These results provide insights into the energetics of intercomponent electron transfer in nitrogenase, suggesting that the energy of protein-protein complex formation is coupled to an increase in the driving force for electron transfer. The results are interpreted in light of the expected changes in the protein environments of the metal centers within the nitrogenase complex. PMID- 9335559 TI - Analysis of some optical properties of a native and reconstituted photosystem II antenna complex, CP29: pigment binding sites can be occupied by chlorophyll a or chlorophyll b and determine spectral forms. AB - The minor photosystem II antenna complex CP29(Lhcb-4) has been reconstituted in vitro with the Lhcb-4 apoprotein, overexpressed in Escherichia coli, and the native pigments. Modulation of the pigment composition during reconstitution yields binding products with markedly different chlorophyll a/b binding ratios even though the total number of bound chlorophylls (a plus b) remains constant at eight. A thermodynamic analysis of steady state absorption and fluorescence spectra demonstrates that all chlorophylls are energetically coupled, while the kinetics of chlorophyll photooxidation indicate that triplet chlorophyll carotenoid coupling is also conserved during pigment binding in vitro. The influence of the chlorophyll a/b binding ratio on the absorption spectra measured at 72 and 300 K is analyzed for the Qy absorption region. Increased chlorophyll b binding leads to large increases in absorption in the 640-660 nm region, while absorption in the 675-690 nm interval decreases markedly. These changes are analyzed in terms of a Gaussian decomposition description in which the eight subbands display a temperature-dependent broadening in agreement with the weak electron-phonon coupling demonstrated for other antenna chlorophyll spectral forms. In this way, we demonstrate that increased chlorophyll b binding leads to increased absorption intensity associated with the subbands at 640, 648, 655, and 660 nm and decreased intensity for the long wavelength subbands at 678 and 684 nm. The wavelength position of all subbands is unchanged. The above data are interpreted to indicate that CP29 has eight chlorophyll binding sites, many or all of which can be occupied by either chlorophyll a or chlorophyll b according to the conditions in which pigment binding occurs. Chlorophyll b absorption is primarily associated with four subbands located at 640, 648, 655, and 660 nm. The invariance of the wavelength position of the absorption bands in recombinant products with different chlorophyll a/b binding stoichiometries is discussed in terms of the mechanism involved in the formation of spectral bands. We conclude that pigment-protein interactions dominate in the determination of spectral heterogeneity with probably only minor effects on absorption associated with pigment-pigment interactions. PMID- 9335560 TI - Linkage between operator binding and dimer to octamer self-assembly of bacteriophage lambda cI repressor. AB - Cooperative binding of the bacteriophage lambda cI repressor dimer to specific sites of the phage operators OR and OL controls the developmental state of the phage. Cooperativity has long been thought to be mediated by self-assembly of repressor dimers to form tetramers which can bind simultaneously to adjacent operators. More recently, we demonstrated that when free repressor dimers self associate in solution, tetramer is an intermediate in a concerted assembly reaction leading to octamer as the predominant higher order species [Senear, D. F., et al. (1993) Biochemistry 32, 6179-6189]. Even as a minority component in the assembly reaction, tetramer can account for pairwise cooperativity. In a similar manner, were it able to bind all three operators simultaneously, octamer could account for three-way cooperativity. In fact, based solely on repressor self-assembly, the naive prediction is that the repressor-OR interactions should be substantially more cooperative than they are. Evidently, there are unfavorable contributions to cooperativity from processes other than repressor self-assembly. Here, we focus on coupling between repressor self-association and operator binding as one possible unfavorable contribution to cooperativity. Sedimentation equilibrium analysis was used to compare the dimer-octamer association reactions of a repressor dimer-OR1 complex and free repressor dimer. Fluorescence anisotropy was used to investigate OR1 binding to free dimers and dimers assembled as higher order species. The results of these experiments indicate a significant and salt-dependent unfavorable contribution generated by such coupling. Since the oligonucleotides used in these experiments are the size of single operator sites, this coupling is mediated by the protein, not by the DNA. This mechanism does not account for an additional, salt-independent, unfavorable contribution which we presume is transmitted via the DNA. Thus, unfavorable contributions generated by structural transitions in both macromolecules serve to moderate the effect of self-association alone. We speculate that this is a general feature of cooperative protein-DNA interactions. PMID- 9335561 TI - Bimolecular DNA triplexes: duplex extensions show implications for H-form DNA stability. AB - H-form DNA has recently been shown to be biologically relevant by its involvement in the process of homologous recombination [Kohwi, Y. , and Panchenko, Y. (1993) Genes Dev. 7, 1766-1778]. A bimolecular DNA triple-stranded structure (triplex) is central to the formation of H-form DNA. Understanding the formation and factors governing the stability of such bimolecular triplexes is necessary to fully elucidate the structure/function relationship of H-form DNA. In this study, we extend known information on bimolecular triplexes by examining the effect of a variable CNC base triad (where N = A, C, T, or G) on a 10 base triad triplex that mimics the triplex motif in H-form DNA. We also examine the effect that a duplex extension of four base pairs has on triplex stability and selectivity for the base N. Results from thermal denaturation experiments indicate that the fully complementary triplex is more stable than its duplex counterpart (DeltaTm = 13 degrees C) and is resistant to degradation by bovine spleen phosphodiesterase for at least 24 h at 10 degrees C. A single-base mismatch in the purine strand of the triplex structure is destabilizing (DeltaTm = approximately 20 degrees C), and all structures containing a mismatch were readily degraded by bovine spleen phosphodiesterase. An extension of four duplex base pairs onto the triplex structure affects the stability of the DNA complex and may have implications relevant to H-form DNA. PMID- 9335562 TI - Miscoding potential of tamoxifen-derived DNA adducts: alpha-(N2 deoxyguanosinyl)tamoxifen. AB - The treatment of tamoxifen, widely used as adjuvant chemotherapy for breast cancer, increases significantly the risk of developing endometrial cancer. The miscoding properties of tamoxifen-derived DNA adducts, alpha-(N2 deoxyguanosinyl)tamoxifens (dG-N2-tamoxifen), have been explored, using an in vitro experimental system to quantify base substitutions and deletions. Site specifically modified oligodeoxynucleotides containing an epimer of trans- and cis-forms of dG-N2-tamoxifens were prepared postsynthetically and used as templates in primer extension reactions catalyzed by mammalian DNA polymerases alpha, beta, and delta. Pol alpha catalyzed incorporation of dCMP and dAMP opposite all four stereoisomers of dG-N2-tamoxifen, accompanied by lesser amounts of dGMP. In contrast, pol delta catalyzed preferential incorporation of dCMP, a correct base, opposite the lesions; one of the trans-forms of dG-N2-tamoxifens only promoted incorporation of dTMP. Using pol beta, preferential incorporation of dCMP, along with small amounts of incorporation of dAMP and dGMP, was detected. One- and two base deletions were also observed with pol alpha and pol beta. The miscoding specificities and frequencies of dG-N2-tamoxifens varied depending on the DNA polymerase used. In addition, with pol alpha and pol beta, large amounts of 5-base deletions were preferentially formed at the cis-forms of dG-N2-tamoxifen, but not at the trans-forms of dG-N2-tamoxifen. We conclude that dG-N2-tamoxifen adducts have high miscoding potentials. PMID- 9335563 TI - Formation and cleavage of a DNA network during in vitro bacteriophage T7 DNA packaging: light microscopy of DNA metabolism. AB - To understand in vivo DNA metabolism, in vitro systems are developed that perform DNA metabolism, while maintaining in vivo (physiological) character. To determine the state of DNA during in vitro physiological metabolism, the present study develops procedures of fluorescence light microscopy for observation of stained DNA molecules during in vitro physiological metabolism in a crude extract of bacteriophage T7-infected cells. The extract inhibits illumination-induced breakage of DNA. The following DNA metabolism remains active for 2-3 min during microscopy: exonuclease-dependent end-to-end joining (concatemerization) of T7 DNA and subsequent cleavage of concatemers. When the T7 gene 3-encoded DNA debranching endonuclease is absent during in vitro T7 DNA concatemerization, DNA progressively partitions to form a continuous, mostly immobile (i.e., no detected Brownian motion) fibrous network that encloses the DNA-depleted solution; presumably because of reduced branching, a less extensive network forms when the gene 3-encoded debranching endonuclease is present. Most strands of the network consist of multiple DNA segments. After a time interval of 5-10 min, the DNA network undergoes cleavage that depends on the presence of both ATP, capsids, and the DNA packaging accessory proteins encoded by genes 18 and 19; multiple cleavages eventually disrupt the continuity of the DNA network. The dependence of the observed cleavage on these factors is explained by the hypothesis that this cleavage is the first of two cleavages known to occur during the packaging of T7 DNA concatemers both in vivo and in vitro. The first cleavage is also known to initiate entry of DNA into a T7 capsid. The cleavage observed here is usually preceded by an approximately 10 s burst of oscillatory motion of the DNA network near the point of eventual cleavage. If the in vivo presence of a similar concatemer-containing DNA network is assumed, requirement for DNA packaging associated release of DNA from this network is a possible explanation for the evolution of a T7 DNA packaging pathway that is initiated by cleavage of a concatemer. PMID- 9335564 TI - Mapping of the residues involved in a proposed beta-strand located in the ferric enterobactin receptor FepA using site-directed spin-labeling. AB - Electron paramagnetic resonance (EPR) site-directed spin-labeling (SDSL) has been used to characterize a proposed transmembrane beta-strand of the Escherichia coli ferric enterobactin receptor, FepA. Each of nine consecutive residues was mutated to cysteine and subsequently labeled with the sulfhydryl-specific spin-label methanethiosulfonate (MTSL) and the purified protein reconstituted into liposomes. Continuous wave (CW) power saturation methods were used to determine exposure of the nitroxide side chains to a series of paramagnetic relaxation agents, including nickel acetylacetonate (NiAA), nickel ethylenediaminediacetate (NiEDDA), chromium oxalate (CROX), and molecular oxygen. The spin-label attached to Q245C, L247C, L249C, A251C, and Y253C had higher collision frequencies with molecular oxygen than with polar relaxation agents, indicating that these sites are exposed to the hydrophobic phase of the lipid bilayer. MTSL bound to residues S246C, E248C, E250C, and G252C had higher collision rates with the polar agents than with oxygen, suggesting that these sites are exposed to the aqueous channel. The alternating periodicity observed with the polar relaxation agents, NiAA and NiEDDA, and in opposite phase with oxygen, is consistent with beta-sheet structure. Depth measurements, based on the reciprocal concentration gradients of NiEDDA and O2 across the bilayer and calibrated for our system with phosphatidylcholine spin-labels, indicated that L249C was nearest the center of the bilayer and that Q245C and Y253C were located just below the bilayer surface in opposite leaflets of the membrane. Thus, we conclude that this approach, through mapping of individual residues, has the capability of defining beta-sheet secondary structure. PMID- 9335565 TI - Resonance Raman, infrared, and EPR investigation on the binuclear site structure of the heme-copper ubiquinol oxidases from Acetobacter aceti: effect of the heme peripheral formyl group substitution. AB - Acetobacter aceti produces two different terminal ubiquinol oxidases (cytochromes a1 and o) depending on the culture conditions. Two types of oxidases share a common protein moiety but with different heme components at the binuclear center (heme A for cytochrome a1 and heme O for cytochrome o). We investigated the structure of the binuclear site of the two oxidases using resonance Raman, Fourier transform-infrared (FT-IR), and EPR spectroscopies to clarify the interactions of heme A formyl group with protein moiety. We found that the overall architecture and the electronic configuration at the binuclear center in the oxidized state seem to be well conserved irrespective of the heme peripheral group at position 8, except for the azide-inhibited state. In contrast, we observed great variations in the C-N stretching frequency and cyanide-binding affinity in the CN-reduced state, in addition to multiple C-O stretching bands in the CO-reduced state. Present and previous studies suggest that the conformational flexibility of the binuclear center in the reduced ligand-bound state may be a common feature among the heme-copper oxidase superfamily. In the CN-reduced state, a hydrogen bond network may be formed among the formyl group, water molecule(s), and the surrounding amino acid residue(s). This network may be very important to maintain proper orientations of the distal amino acid residues and/or the CuB1+ ion relative to the cyanide ion bound to the ferrous heme iron and could play a critical role for the high affinity in cyanide binding. PMID- 9335566 TI - Backbone dynamics and structural characterization of the partially folded A state of ubiquitin by 1H, 13C, and 15N nuclear magnetic resonance spectroscopy. AB - Structure and dynamics of the partially folded A state of ubiquitin in a 60%/40% methanol/water mixture at pH 2 was studied by two- and three-dimensional nuclear magnetic resonance spectroscopy (NMR) using fully 13C,15N-labeled ubiquitin. Complete backbone 13CO, 13Calpha, 15N, and 1HN assignment was achieved. 13CO and 13Calpha chemical shifts and 1H-1H nuclear Overhauser enhancement (NOE) connectivities indicate different behavior for the N-terminal and the C-terminal halves of the protein. In the N-terminal half of the A state, comprising the antiparallel beta-sheet and the central alpha-helix, the native secondary structural elements are largely conserved. The C-terminal half, which is in the native form rich in beta-strand character, undergoes a methanol-induced transition to a dynamic state with a uniformly high propensity for helical structure. This behavior is also reflected in backbone 15N relaxation data, indicating the presence of three loosely coupled secondary structural segments with enhanced internal mobility as compared to the native state. PMID- 9335567 TI - Metal-dependent conformers of the periplasmic ferric ion binding protein. AB - One of the better understood structural correlates of Fe3+ binding by the transferrins is the conformational shift demonstrated by both lobes. FbpA, a prokaryotic protein involved in periplasmic iron transport, has previously been shown to be structurally and functionally homologous to the transferrins. Similar to each individual lobe of the transferrins, it is hypothesized that FbpA exists in two distinct conformations depending on whether metal is bound. Evidence for these changes is provided by the differential susceptibility of FbpA to trypsin digestion. Binding of Fe3+ by FbpA significantly decreases the ability of trypsin to digest wild-type protein. Construction of a null binding mutant, Tyr195Ile, confirms that protein "locked" in the apo-conformation is similarly susceptible to trypsin. This mutant also marks the initial characterization of an FbpA molecule unable to bind iron, suggesting that the Tyr195 residue is directly involved in iron binding. Other FbpA mutants which do bind iron show moderate resistance to digestion which suggests that they remain in the holo-protein conformation when binding Fe3+. The conformational states of FbpA may have important implications in protein-protein recognition during transport of Fe3+ between membranes, and may explain how these proteins function in the context of periplasm-to-cytosol Fe3+ transport. PMID- 9335568 TI - Apolipoprotein B binding to microsomal triglyceride transfer protein decreases with increases in length and lipidation: implications in lipoprotein biosynthesis. AB - Microsomal triglyceride transfer protein (MTP), a heterodimer of 97 kDa and protein disulfide isomerase, is required for the assembly of apolipoprotein B (apoB)-containing triglyceride-rich lipoproteins. These proteins have been shown to interact with each other during early stages of lipoprotein biosynthesis. Our studies indicated that binding between apoB and heterodimeric MTP was of high affinity (Kd 10-30 nM) due to ionic interactions. In contrast to MTP, protein disulfide isomerase alone interacted very poorly with lipoproteins, indicating the importance of the heterodimer in these bindings. Preincubation of lipoproteins with detergents enhanced their interaction with MTP. Native VLDL bound poorly to MTP, but its preincubation with Tween-20 resulted in significantly increased binding to MTP. Furthermore, binding of LDL was enhanced by preincubation with taurocholate, indicating that partial delipidation of apoB containing lipoproteins results in increased binding to MTP. Subsequently, attempts were made to study interactions between C-terminally truncated apoB polypeptides and MTP. Binding of all the polypeptides to MTP was enhanced in the presence of taurocholate. Comparisons revealed that the binding of different apoB polypeptides to MTP was in the order of apoB18 > apoB28 > apoB42 > apoB100. These studies indicated that optimum interactions occur between apoB18 and MTP, and that the increase in apoB length beyond apoB18 has a negative effect on these interactions. Since apoB18 does not assemble triglyceride-rich lipoproteins, these studies suggest that apoB may interact with MTP before its lipidation. It is proposed that steps in lipoprotein biosynthesis may be dictated by the sequential display of different functional domains on the apoB polypeptide. PMID- 9335569 TI - Regeneration studies of an analog of ribonuclease A missing disulfide bonds 65-72 and 40-95. AB - Mutants of bovine pancreatic ribonuclease A (RNase A) that contain four of the eight cysteine residues found in the wild-type protein were prepared. Cysteine residues 40, 65, 72, and 95 were replaced by serine to form [C40S,C65S,C72S,C95S] RNase A or by alanine to form [C40A,C65A,C72A,C95A] RNase A, which contain the following four cysteine residues: 26, 58, 84, and 110. The substitutions resulted in deletion of wild-type disulfide bonds, 65-72 and 40-95. These mutants were prepared to investigate interactions that may be important for the folding and unfolding of the wild-type protein. The mutant protein was expressed and purified in an unfolded sulfonated form. Upon regeneration of the native form from the reduced mutant with DTTox, all three of the possible two-disulfide pairings, including the native one, formed. One-dimensional 1H NMR spectra demonstrated that the conformations of these three species are similar and are predominantly disordered; however, there is evidence of local structure in the vicinity of one histidine residue. It was also shown that disulfide pairing is not completely random and that both entropic factors and enthalpic interactions contribute to the formation of the native-disulfide bonds. The presence of more than a statistical population of native-disulfide pairings indicates that specific local interactions present in the reduced protein direct the preferential formation of native-disulfide bonds in the two-disulfide mutant. PMID- 9335570 TI - Arginine294 is essential for the inhibition of Anabaena PCC 7120 ADP-glucose pyrophosphorylase by phosphate. AB - Treatment of ADP-glucose pyrophosphorylase (EC 2.7.7.27) from the cyanobacterium Anabaena PCC 7120 with phenylglyoxal in 50 mM Hepes, pH 8.0, at 25 degrees C resulted in a time- and concentration-dependent loss of enzyme activity. Phosphate, the inhibitor, protected the enzyme from inactivation most effectively, while 3-P-glycerate, fructose-1,6-P2, pyridoxal-P, and ATP plus magnesium were also good protectors. After incubation with 2 mM phenylglyoxal for 1 h, the modified enzyme had a 10-fold lower apparent affinity for phosphate in the absence of the activator, 3-P-glycerate, than that of the wild-type enzyme. This result has implicated the involvement of an arginine residue at the allosteric sites, most probably the inhibitor-binding site, of ADP-glucose pyrophosphorylase from the cyanobacterium Anabaena PCC 7120. In order to identify the arginine residue, five arginine residues, which are conserved in all higher plant and cyanobacterial enzymes but not in enteric bacterial enzymes, were individually converted to alanine by site-directed mutagenesis. The mutant enzymes, R66A, R105A, R294A, and R385A, were purified, and the properties of these mutants were compared with the wild-type enzyme. Substitution of arginine294 with alanine resulted in an enzyme with more than 100-fold or 40-fold lower affinity for the inhibitor, phosphate, in the absence or presence of 3-P glycerate, respectively. This mutation had no or lesser impact on the kinetic constants for the substrates and the activator, 3-P-glycerate. PMID- 9335571 TI - Stoichiometry of the interaction of prostaglandin H synthase with substrates. AB - Prostaglandin H synthase (PGHS) catalyzes both peroxidase and cyclooxygenase reactions. Resolution of several current issues regarding the PGHS catalytic mechanism hinges on the stoichiometry of the reaction of PGHS with hydroperoxide, fatty acid, and oxygen. The dependence of wide-doublet tyrosyl radical accumulation in PGHS isoform 1 on hydroperoxide stoichiometry, has been determined; this catalytically active radical is formed efficiently at stoichiometries DL) is limited by the rate of disruption of the hydrophobic core in MS. Equilibrium spectroscopic measurements by near-IR and Soret absorbance, fluorescence, and circular dichroism showed that DL has native like helical secondary structure, but shows no evidence for specific tertiary interactions. This lipid-denatured equilibrium state (DL) is clearly more extensively unfolded than the A-state in solution, but is distinct from the unfolded protein in water (UW), which has no stable secondary structure. PMID- 9335577 TI - Dobzhansky's genetics and the origin of species: is it still relevant? PMID- 9335576 TI - Preparative induction and characterization of L-antithrombin: a structural homologue of latent plasminogen activator inhibitor-1. AB - The inhibitory mechanism of the serpin family of serine protease inhibitors is characterized by a remarkable degree of conformational flexibility. Various conformational states have been elucidated by X-ray crystallography and indicate that the inhibitory loop, the central A-beta-sheet, and the outside edge of the C beta-sheet are particularly mobile. However, no crystal structure of a serpin enzyme complex is yet available, and the likely nature of the protease-complexed serpin remains for biochemical and biophysical researchers to examine. Here, we show that the biochemical induction of the latent state of antithrombin is slow relative to polymer formation, and infer that this may reflect structural features that are important for the regulation of the initial docking and subsequent locking of serpins with cognate proteases. L-Antithrombin was induced by incubation of native antithrombin at 60 degrees C for 10 h in the presence of citrate to prevent polymerization. L-Antithrombin was more stable to denaturation by both heat and urea than native antithrombin. Whereas native antithrombin formed binary complexes with synthetic peptide homologues of the inhibitory loop, biochemically induced L-antithrombin did not, indicating that the inhibitory loop of L-antithrombin is probably fully inserted into the A-beta-sheet as in the crystal structure. This was confirmed by limited proteolysis studies which demonstrated that the inhibitory loop of L-antithrombin could not be cleaved by five proteases which do cleave the loop of native antithrombin. The limited proteolysis studies also indicated that the "gate" region (residues 236-248) of the biochemically induced L-antithrombin was in a conformation substantially different from that of the native antithrombin. This again is similar to L antithrombin in the crystal structure in which the gate has "opened" away from the body of the molecule by a rotation of 24 degrees to facilitate the relocation of strand 1C from its ordered position in the C-beta-sheet to a disordered surface loop. At 60 degrees C in the absence of citrate, antithrombin (and other serpins) rapidly polymerizes. In the presence of citrate, the formation of L antithrombin is slow and increases with time, indicating that the inhibition of polymer formation by citrate allows the time necessary for the much slower formation of the L form. We therefore suggest that L-antithrombin formation is a two-step process: an initial rapid conformational change, probably including partial incorporation of the reactive loop into the A-sheet (as in the active molecule in the crystal structure) and displacement of s1C from the C-beta-sheet which supports polymer formation, and a much slower transition to complete loop insertion within the A-beta-sheet. It is likely that both the first rapid transitional step and the structural features that impose resistance to the second more extensive conformational change reflect the optimization of the unique inhibitory function in the serpins. PMID- 9335578 TI - Roles for lambda Orf and Escherichia coli RecO, RecR and RecF in lambda recombination. AB - Bacteriophage lambda lacking its Red recombination functions requires either its own gene product, Orf, or the product of Escherichia coli's recO, recR and recF genes (RecORF) for efficient recombination in recBC sbcB sbcC mutant cells (the RecF pathway). Phage crosses under conditions of a partial block to DNA replication have revealed the following: (1) In the presence of Orf, RecF pathway recombination is similar to lambda Red recombination; (2) Orf is necessary for focusing recombination toward the right end of the chromosome as lambda is conventionally drawn; (3) RecORF-mediated RecF pathway recombination is not focused toward the right end of the chromosome, which may indicate that RecORF travels along the DNA; (4) both Orf- and RecORF-mediated RecF pathway recombination are stimulated by DNA replication; and (5) low level recombination in the simultaneous absence of Orf and RecORF may occur by a break-copy mechanism that is not initiated by a double strand break. Models for the roles of Orf and RecO, RecR and RecF in recombination are presented. PMID- 9335579 TI - "Break copy" duplication: a model for chromosome fragment formation in Saccharomyces cerevisiae. AB - Introduction of a chromosome fragmentation vector (CFV) into the budding yeast Saccharomyces cerevisiae results in a targeted homologous recombination event that yields an independently segregating chromosome fragment (CF) and an alteration in the strain's karyotype. Fragmentation with an acentric CFV directed in a centromere-proximal orientation generates a CF that contains all sequences proximal to the targeting segment and results in loss of the endogenous targeted chromosome to yield a 2N-1 + CF karyotype. In contrast, fragmentation with a centric CFV directed in a centromere-distal orientation generates a CF that contains all sequences distal to the targeting segment and retention of the endogenous targeted chromosome to yield a 2N + CF karyotype. We have termed this phenomenon "break copy" duplication. Using yeast strains in which the centromere had been transposed to a new location, it was demonstrated that the centromere inhibited break copy duplication. These data suggest that CF formation is the product of an unscheduled DNA replication event initiated by the free end of the CFV and is analogous to a "half" double-strand break gap-repair reaction. We suggest that break copy duplication may have evolved as a mechanism for maintenance of ploidy following DNA breakage. PMID- 9335580 TI - The isolation and characterization of Saccharomyces cerevisiae mutants that constitutively express purine biosynthetic genes. AB - In response to an external source of adenine, yeast cells repress the expression of purine biosynthesis pathway genes. To identify necessary components of this signalling mechanism, we have isolated mutants that are constitutively active for expression. These mutants were named bra (for bypass of repression by adenine). BRA7 is allelic to FCY2, the gene encoding the purine cytosine permease and BRA9 is ADE12, the gene encoding adenylosuccinate synthetase. BRA6 and BRA1 are new genes encoding, respectively, hypoxanthine guanine phosphoribosyl transferase and adenylosuccinate lyase. These results indicate that uptake and salvage of adenine are important steps in regulating expression of purine biosynthetic genes. We have also shown that two other salvage enzymes, adenine phosphoribosyl transferase and adenine deaminase, are involved in activating the pathway. Finally, using mutant strains affected in AMP kinase or ribonucleotide reductase activities, we have shown that AMP needs to be phosphorylated to ADP to exert its regulatory role while reduction of ADP into dADP by ribonucleotide reductase is not required for adenine repression. Together these data suggest that ADP or a derivative of ADP is the effector molecule in the signal transduction pathway. PMID- 9335581 TI - Rules of donor preference in saccharomyces mating-type gene switching revealed by a competition assay involving two types of recombination. AB - Mating type (MAT) switching in Saccharomyces cerevisiae is initiated by a double strand break (DSB) created at MAT by HO endonuclease. MATa cells activate the entire left arm of chromosome III; thus MATa preferentially recombines with the silent donor HML. In contrast, MAT alpha cells inactivate the left arm, including HML, and thus preferentially recombine with HMR, 100 kb to the right of MAT. We present a novel competition assay, in which the DSB at MAT can be repaired either by MAT switching or by single-strand annealing (SSA) between two URA3 genes flanking MAT. With preferred donors, MATa or MAT alpha switching occurs 65-70% of the time in competition with SSA. When HML is deleted, 40% of MATa cells recombine with the "wrong" donor HMR; however, when HMR is deleted, only 18% of MAT alpha cells recombine with HML. In interchromosomal switching, with donors on chromosome III and MAT on chromosome V, MATa retains its strong preference for HML and switching is efficient, when the chromosome III recombination enhancer is present. However, MAT alpha donor preference is lost and interchromosomal switching is very inefficient. These experiments demonstrate the utility of using competition between two outcomes to measure the relative efficiency of recombination. PMID- 9335582 TI - Suppressors of the ndc10-2 mutation: a role for the ubiquitin system in Saccharomyces cerevisiae kinetochore function. AB - We have isolated a new conditional-lethal mutation, ndc10-2, in the NDC10/CBF2/CTF14 gene that encodes the 110-kD subunit of the Saccharomyces cerevisiae CBF3 kinetochore complex. At the restrictive temperature of 37 degrees, ndc10-2 cells are able to assemble anaphase spindles, but fail to segregate their DNA, consistent with a defect in kinetochore function. To identify other factors that play a role in kinetochore assembly or function, we isolated both dosage and second site suppressors of the ndc10-2 mutation. These screens identified UBC6 as a dosage suppressor, and mutations in UBC6 and UBC7 as second-site suppressors of ndc10-2 heat sensitivity. Both UBC6 and UBC7 encode ubiquitin-conjugating enzymes that function in ubiquitin-mediated protein degradation. Furthermore, overexpression of a mutant ubiquitin suppresses the ndc10-2 mutation. These results implicate the ubiquitin system in the regulation of ndc10-2 function and suggest a role for the ubiquitin system in kinetochore function. PMID- 9335583 TI - Identification of genes controlling growth polarity in the budding yeast Saccharomyces cerevisiae: a possible role of N-glycosylation and involvement of the exocyst complex. AB - The regulation of secretion polarity and cell surface growth during the cell cycle is critical for proper morphogenesis and viability of Saccharomyces cerevisiae. A shift from isotropic cell surface growth to polarized growth is necessary for bud emergence and a repolarization of secretion to the bud neck is necessary for cell separation. Although alterations in the actin cytoskeleton have been implicated in these changes in secretion polarity, clearly other cellular systems involved in secretion are likely to be targets of cell cycle regulation. To investigate mechanisms coupling cell cycle progression to changes in secretion polarity in parallel with and downstream of regulation of actin polarization, we implemented a screen for mutants defective specifically in polarized growth but with normal actin cytoskeleton structure. These mutants fell into three classes: those partially defective in N-glycosylation, those linked to specific defects in the exocyst, and a third class neither defective in glycosylation nor linked to the exocyst. These results raise the possibility that changes in N-linked glycosylation may be involved in a signal linking cell cycle progression and secretion polarity and that the exocyst may have regulatory functions in coupling the secretory machinery to the polarized actin cytoskeleton. PMID- 9335584 TI - Large scale identification of genes involved in cell surface biosynthesis and architecture in Saccharomyces cerevisiae. AB - The sequenced yeast genome offers a unique resource for the analysis of eukaryotic cell function and enables genome-wide screens for genes involved in cellular processes. We have identified genes involved in cell surface assembly by screening transposon-mutagenized cells for altered sensitivity to calcofluor white, followed by supplementary screens to further characterize mutant phenotypes. The mutated genes were directly retrieved from genomic DNA and then matched uniquely to a gene in the yeast genome database. Eighty-two genes with apparent perturbation of the cell surface were identified, with mutations in 65 of them displaying at least one further cell surface phenotype in addition to their modified sensitivity to calcofluor. Fifty of these genes were previously known, 17 encoded proteins whose function could be anticipated through sequence homology or previously recognized phenotypes and 15 genes had no previously known phenotype. PMID- 9335585 TI - Essential functional interactions of SAGA, a Saccharomyces cerevisiae complex of Spt, Ada, and Gcn5 proteins, with the Snf/Swi and Srb/mediator complexes. AB - The Saccharomyces cerevisiae transcription factor Spt20/Ada5 was originally identified by mutations that suppress Ty insertion alleles and by mutations that suppress the toxicity caused by Gal4-VP16 overexpression. Here we present evidence for physical associations between Spt20/Ada5 and three other Spt proteins, suggesting that they exist in a complex. A related study demonstrates that this complex also contains the histone acetyltransferase, Gcn5, and Ada2. This complex has been named SAGA (Spt/Ada/Gcn5 acetyltransferase). To identify functions that genetically interact with SAGA, we have screened for mutations that cause lethality in an spt20 delta/ada5 delta mutant. Our screen identified mutations in SNF2, SIN4, and GAL11. These mutations affect two known transcription complexes: Snf/Swi, which functions in nucleosome remodeling, and Srb/mediator, which is required for regulated transcription by RNA polymerase II. Systematic analysis has demonstrated that spt20 delta/ada5 delta and spt7 delta mutations cause lethality with every snf/swi and srb/mediator mutation tested. Furthermore, a gcn5 delta mutation causes severe sickness with snf/swi mutations, but not with srb/mediator mutations. These findings suggest that SAGA has multiple activities and plays critical roles in transcription by RNA polymerase II. PMID- 9335586 TI - Genetic interactions between a pep7 mutation and the PEP12 and VPS45 genes: evidence for a novel SNARE component in transport between the Saccharomyces cerevisiae Golgi complex and endosome. AB - The PEP7 gene from Saccharomyces cerevisiae encodes a 59-kD hydrophilic polypeptide that is required for transport of soluble vacuolar hydrolase precursors from the TGN to the endosome. This study presents the results of a high-copy suppression analysis of pep7-20 mutant phenotypes. This analysis demonstrated that both VPS45 and PEP12 are allele-specific high-copy suppressors of pep7-20 mutant phenotypes. Overexpression of VPS45 was able to completely suppress the Zn2+ sensitivity and partially suppress the carboxypeptidase Y deficiency. Overexpression of PEP12 was able to do the same, but to a lesser extent. Vps45p and Pep12p are Sec1p and syntaxin (t-SNARE) homologues, respectively, and are also thought to function in transport between the TGN and endosome. Two additional vacuole pathway SNARE complex homologues, Vps33p (Sec1p) and Pth1p (syntaxin), when overexpressed, were unable to suppress pep7-20 or any other pep7 allele, further supporting the specificity of the interactions of pep7 20 with PEP12 and VPS45. Because several other vesicle docking/fusion reactions take place in the cell without discernible participation of Pep7p homologues, we suggest that Pep7p is a step-specific regulator of docking and/or fusion of TGN derived transport vesicles onto the endosome. PMID- 9335587 TI - Mutational analysis of STE5 in the yeast Saccharomyces cerevisiae: application of a differential interaction trap assay for examining protein-protein interactions. AB - Ste5 is essential for the yeast mating pheromone response pathway and is thought to function as a scaffold that organizes the components of the mitogen-activated protein kinase (MAPK) cascade. A new method was developed to isolate missense mutations in Ste5 that differentially affect the ability of Ste5 to interact with either of two MAPK cascade constituents, the MEKK (Ste11) and the MEK (Ste7). Mutations that affect association with Ste7 or with Ste11 delineate discrete regions of Ste5 that are critical for each interaction. Co-immunoprecipitation analysis, examining the binding in vitro of Ste5 to Ste11, Ste7, Ste4 (G protein beta subunit), and Fus3 (MAPK), confirmed that each mutation specifically affects the interaction of Ste5 with only one protein. When expressed in a ste5 delta cell, mutant Ste5 proteins that are defective in their ability to interact with either Ste11 or Ste7 result in a markedly reduced mating proficiency. One mutation that clearly weakened (but did not eliminate) interaction of Ste5 with Ste7 permitted mating at wild-type efficiency, indicating that an efficacious signal is generated even when Ste5 associates with only a small fraction of (or only transiently with) Ste7. Ste5 mutants defective in association with Ste11 or Ste7 showed strong interallelic complementation when co-expressed, suggesting that the functional form of Ste5 in vivo is an oligomer. PMID- 9335588 TI - Specialized Rap1p/Gcr1p transcriptional activation through Gcr1p DNA contacts requires Gcr2p, as does hyperphosphorylation of Gcr1p. AB - The multifunctional regulatory factor Rap1p of Saccharomyces cerevisiae accomplishes one of its tasks, transcriptional activation, by complexing with Gcr1p. An unusual feature of this heteromeric complex is its apparent capacity to contact simultaneously two adjacent DNA elements (UASRPG and the CT box, bound specifically by Rap1p and Gcr1p, respectively). The complex can activate transcription through isolated UASRPG but not CT elements. In promoters that contain both DNA signals its activity is enhanced, provided the helical spacing between the two elements is appropriate; this suggests that at least transient DNA loop formation is involved. We show here that this CT box-dependent augmentation of Rap1p/Gcr1p activation requires the presence of a third protein Gcr2p; the Gcr2- growth defect appears to result from a genome-wide loss of the CT box effect. Interestingly, a hyperphosphorylated form of Gcr1p disappears in delta gcr2 cells but reappears if they harbor a doubly point-mutated GCR1 allele that bypasses the Gcr2- growth defect. Gcr2p therefore appears to induce a conformation change in Gcr1p and/or stimulate its hyperphosphorylation; one or both of these effects can be mimicked in the absence of GCR2 by mutation of GCR1. This improved view of Rap1p/Gcr1p/Gcr2p function reveals a new aspect of eukaryotic gene regulation: modification of an upstream activator, accompanied by at least transient DNA loop formation, mediates its improved capacity to activate transcription. PMID- 9335589 TI - Genetic and environmental factors affecting the de novo appearance of the [PSI+] prion in Saccharomyces cerevisiae. AB - It has previously been shown that yeast prion [PSI+] is cured by GuHCl, although reports on reversibility of curing were contradictory. Here we show that GuHCl treatment of both [PSI+] and [psi-] yeast strains results in two classes of [psi ] derivatives: Pin+, in which [PSI+] can be reinduced by Sup35p overproduction, and Pin-, in which overexpression of the complete SUP35 gene does not lead to the [PSI+] appearance. However, in both Pin+ and Pin- derivatives [PSI+] is reinduced by overproduction of a short Sup35p N-terminal fragment, thus, in principle, [PSI+] curing remains reversible in both cases. Neither suppression nor growth inhibition caused by SUP35 overexpression in Pin+ [psi-] derivatives are observed in Pin- [psi-] derivatives. Genetic analyses show that the Pin+ phenotype is determined by a non-Mendelian factor, which, unlike the [PSI+] prion, is independent of the Sup35p N-terminal domain. A Pin- [psi-] derivative was also generated by transient inactivation of the heat shock protein, Hsp104, while [PSI+] curing by Hsp104 overproduction resulted exclusively in Pin+ [psi-] derivatives. We hypothesize that in addition to the [PSI+] prion-determining domain in the Sup35p N-terminus, there is another self-propagating conformational determinant in the C-proximal part of Sup35p and that this second prion is responsible for the Pin+ phenotype. PMID- 9335590 TI - The role of Gcr1p in the transcriptional activation of glycolytic genes in yeast Saccharomyces cerevisiae. AB - To study the interdependence of Gcr1p and Rap1p, we prepared a series of synthetic regulatory sequences that contained various numbers and combinations of CT-boxes (Gcr1p-binding sites) and RPG-boxes (Rap1p-binding sites). The ability of the synthetic oligonucleotides to function as regulatory sequences was tested using an ENO1-lacZ reporter gene. As observed previously, synthetic oligonucleotides containing both CT- and RPG-boxes conferred strong UAS activity. Likewise, a lone CT-box did not show any UAS activity. By contrast, oligonucleotides containing tandem Ct-boxes but no RPG-box conferred strong promoter activity. This UAS activity was not dependent on position or orientation of the oligonucleotides in the 5' noncoding region. However, it was dependent on both GCR1 and GCR2. These results suggest that the ability of Gcr1p to bind Gcr1p binding sites in vivo is not absolutely dependent on Rap1p. Eleven independent mutants of GCR1 were isolated that conferred weak UAS activity to a single CT box. Five mutants has single mutations in Gcr1p's DNA-binding domain and displayed slightly higher affinity for the CT-box. These results support the hypothesis that Gcr1p and Gcr2p play the central role in glycolytic gene expression and that the function of Rap1p is to facilitate the binding of Gcr1p to its target. PMID- 9335591 TI - DMC1 functions in a Saccharomyces cerevisiae meiotic pathway that is largely independent of the RAD51 pathway. AB - Meiotic recombination in the yeast Saccharomyces cerevisiae requires two similar recA-like proteins, Dmc1p and Rad51p. A screen for dominant meiotic mutants provided DMC1-G126D, a dominant allele mutated in the conserved ATP-binding site (specifically, the A-loop motif) that confers a null phenotype. A recessive null allele, dmc1-K69E, was isolated as an intragenic suppressor of DMC1-G126D. Dmc1 K69Ep, unlike Dmc1p, does not interact homotypically in a two-hybrid assay, although it does interact with other fusion proteins identified by two-hybrid screen with Dmc1p. Dmc1p, unlike Rad51p, does not interact in the two-hybrid assay with Rad52p or Rad54p. However, Dmc1p does interact with Tid1p, a Rad54p homologue, with Tid4p, a Rad16p homologue, and with other fusion proteins that do not interact with Rad51p, suggesting that Dmc1p and Rad51p function in separate, though possibly overlapping, recombinational repair complexes. Epistasis analysis suggests that DMC1 and RAD51 function in separate pathways responsible for meiotic recombination. Taken together, our results are consistent with a requirement for DMC1 for meiosis-specific entry of DNA double-strand break ends into chromatin. Interestingly, the pattern on CHEF gels of chromosome fragments that result from meiotic DNA double-strand break formation is different in DMC1 mutant strains from that seen in rad50S strains. PMID- 9335592 TI - High frequency cDNA recombination of the saccharomyces retrotransposon Ty5: The LTR mediates formation of tandem elements. AB - Retroelement cDNA can integrate into the genome using the element-encoded integrase or it can recombine with preexisting elements using the recombination system of the host. Recombination is a particularly important pathway for the yeast retrotransposon Ty5 and accounts for approximately 30% of the putative transposition events when a homologous substrate is carried on a plasmid and approximately 7% when the substrate is located at the chromosomal URA3 locus. Characterization of recombinants revealed that they are either simple replacements of the marker gene tandem elements. Using an assay system in which the donor element and recombination substrates are separated, we found that the long terminal repeats (LTRs) are critical for tandem element formation. LTR containing substrates generate tandem elements at frequencies more than 10-fold higher than similarly sized internal Ty5 sequences. Internal sequences, however, facilitate tandem element formation when associated with an LTR, and there is a linear relationship between frequencies of tandem element formation and the length of LTR-containing substrates. We propose that recombination is initiated between the LTRs of the cDNA and substrate and that internal sequences promote tandem element formation by facilitating sequence alignment. Because of its location in subtelomeric regions, recombinational amplification of Ty5 may contribute to the organizations of chromosome ends. PMID- 9335593 TI - Mutations in GSF1 and GSF2 alter glucose signaling in Saccharomyces cerevisiae. AB - One function of the Saccharomyces cerevisiae Snf1 protein kinase is to relieve glucose repression of SUC, GAL, and other genes in response to glucose depletion. To identify genes that regulate Snf1 kinase activity, we have selected mutants that inappropriately express a SUC2promoter::HIS3 gene fusion when grown in glucose and that require Snf1 function for this phenotype. Mutations representing two new complementation groups (gsf1 and gsf2) were isolated. gsf1 mutations affect two distinct responses to glucose: the Snf1-regulated glucose repression of SUC2 and GAL10 transcription and the Snf1-independent induction by glucose of HXT1 transcription. gsf2 mutations relieve glucose repression of SUC2 and GAL10 transcription and, in combination with snf1 delta, cause an extreme slow growth phenotype. The GSF2 gene was cloned by complementation of the gsf2-1 snf1 delta slow growth phenotype and encodes a previously uncharacterized 46kD protein. PMID- 9335594 TI - Mating-type genes from the homothallic fungus Sordaria macrospora are functionally expressed in a heterothallic ascomycete. AB - Homokaryons from the homothallic ascomycte Sordaria macrospora are able to enter the sexual pathway and to form fertile fruiting bodies. To analyze the molecular basis of homothallism and to elucidate the role of mating-products during fruiting body development, we cloned and sequenced the entire S. macrospora mating-type locus. Comparison of the Sordaria mating-type locus with mating-type idiomorphs from the heterothallic ascomycetes Neurospora crassa and Podospora anserina revealed that sequences from both idiomorphs (A/a and mat-/mat+, respectively) are contiguous in S. macrospora. DNA sequencing of the S. macrospora mating-type region allowed the identification of four open reading frames (ORFs), which were termed Smt-a1, SmtA-1, SmtA-2 and SmtA-3. While Smt-a1, SmtA-1, and SmtA-2 show strong sequence similarities with the corresponding N. crassa mating-type ORFs, SmtA-3 has a chimeric character. It comprises sequences that are similar to the A and a mating-type idiomorph from N. crassa. To determine functionality of the S. macrospora mating-type genes, we show that all ORFs are transcriptionally expressed. Furthermore, we transformed the S. macrospora mating-type genes into mat- and mat+ strains of the closely related heterothallic fungus P. anserina. The transformation experiments show that mating type genes from S. macrospora induce fruiting body formation in P. anserina. PMID- 9335595 TI - A mutation in an HSP90 gene affects the sexual cycle and suppresses vegetative incompatibility in the fungus Podospora anserina. AB - Vegetative incompatibility is widespread in fungi but its molecular mechanism and biological function are still poorly understood. A way to study vegetative incompatibility is to investigate the function of genes whose mutations suppress this phenomenon. In Podospora anserina, these genes are known as mod genes. In addition to suppressing vegetative incompatibility, mod mutations cause some developmental defects. This suggests that the molecular mechanisms of vegetative incompatibility and development pathway are interconnected. The mod-E1 mutation was isolated as a suppressor of the developmental defects of the mod-D2 strain. We show here that mod-E1 also partially suppresses vegetative incompatibility, strengthening the link between development and vegetative incompatibility. mod-E1 is the first suppressor of vegetative incompatibility characterized at the molecular level. It encodes a member of the Hsp90 family, suggesting that development and vegetative incompatibility use common steps of a signal transduction pathway. The involvement of mod-E in the sexual cycle has also been further investigated. PMID- 9335596 TI - Homology-dependent silencing of the SC3 gene in Schizophyllum commune. AB - After introduction of extra copies of the SC3 hydrophobin gene into a wild-type strain of Schizophyllum commune, gene silencing was observed acting on both endogenous and introduced SC3 genes in primary vegetative transformants. Nuclear run-on experiments indicated that silencing acted at the transcriptional level. Southern analysis revealed that cytosine methylation of genomic DNA occurred. Moreover, SC3 silencing was suppressed by exposure to 5-azacytidine during growth. After growth of SC3-suppressed colonies from homogenized mycelium or from colonies stored at 4 degrees, SC3 transcription was restored. However, after prolonged growth SC3 silencing was again observed. Introduction of a promoterless SC3 fragment into wild type gave less SC3 silencing. PMID- 9335597 TI - Characterization of revertants of unc-93(e1500) in Caenorhabditis elegans induced by N-ethyl-N-nitrosourea. AB - Phenotypic reversion of the rubber-band, muscle-defective phenotype conferred by unc-93(e1500) was used to determine the utility of N-ethyl-N-nitrosourea (ENU) as a mutagen for genetic research in Caenorhabditis elegans. In this system, ENU produces revertants at a frequency of 3 x 10(-4), equivalent to that of the commonly used mutagen, EMS. The gene identity of 154 ENU-induced revertants shows that the distribution of alleles between three possible suppressor genes differs from induced by EMS. A higher percentage of revertants are alleles of unc-93 and many fewer are alleles of sup-9 and sup-10. Three revertants complement the three known suppressor genes; they may therefore identify a new gene product(s) involved in this system of excitation-contraction coupling in C. elegans. Molecular characterization of putative unc-93 null alleles reveals that the base changes induced by ENU are quite different from those induced by EMS; specifically we see an increased frequency of A/T-->G/C transitions. The frequency of ENU-induced intragenic deletions is found to be 13%. We suggest that ENU, at concentrations below 5 mM, will be a superior mutagen for studies of protein function in C. elegans. PMID- 9335598 TI - Balancing selection on electrophoretic variation of phosphoglucose isomerase in two species of field cricket: Gryllus veletis and G. offnsylvanicus. AB - Two species of crickets, Gryllus veletis and G. pennsylvanicus, share six electrophoretic mobility classes for the enzyme phosphoglucose isomerase (PGI), despite evidence from other genetic markers that the two species are not closely related within eastern North American field crickets. Moreover, the frequencies of the two most common PGI electrophoretic classes (PGI-100 and PGI-65) covary in sympatric populations of these species in the eastern United States, suggesting that PGI may be subject to trans-specific balancing selection. To determine the molecular basis of the electrophoretic variation, we characterized the DNA sequence of the Pgi gene from 29 crickets (15 G. veletis and 14 G. pennsylvanicus). Amino acid substitutions that distinguish the electrophoretic classes are not the same in the two species, and there is no evidence that specific replacement substitutions represent trans-specific polymorphism. In particular, the amino acids that diagnose the PGI-65 allele relative to the PGI 100 allele differ both between G. veletis and G. pennsylvanicus and within G. pennsylvanicus. The heterogeneity among electrophoretic classes that covary in sympatric populations coupled with analysis of patterns of nucleotide variation suggest that Pgi is not evolving neutrally. Instead, the data are consistent with balancing selection operating on an emergent property of the PGI protein. PMID- 9335599 TI - Drosophila immunity: analysis of larval hemocytes by P-element-mediated enhancer trap. AB - Our aim was to identify new genes involved in the cellular aspects of defense mechanism of Drosophila, as well as in melanotic tumor formation processes that are linked to blood cell disregulation. We have screened 1341 enhancer detector fly lines for expression of the lacZ reporter gene in larval hemocytes at the end of the third instar. We have selected 21 lines in which we observed a reproducible lacZ expression in blood cells. These lines were classified according to the subsets of hemocytes in which lacZ was expressed, and we identified five lines that can be used as lamellocyte markers. Three lines were selected for further analysis. The first exhibited strong lacZ expression in all lamellocytes. The second expressed lacZ in plasmatocytes and lamellocytes, and exhibited a melanotic tumor phenotype in larvae homozygous for the insertion. A third line showed a striking insertion-linked phenotype of melanized lymph glands (the hematopoietic organ), which resulted in the total absence of circulating hemocytes in the mutant larvae. We anticipate that this mutation, which we named domino, will prove a useful tool in the analysis of the role of hemocytes during the various aspects of immune response and melanotic tumor formation. PMID- 9335600 TI - The sex-ratio trait in Drosophila simulans: genetic analysis of distortion and suppression. AB - The sex-ratio trait described in several Drosophila species is a type of naturally occurring X-linked meiotic drive that causes males bearing a sex-ratio X chromosome to produce progenies with a large excess of females. We have previously reported the occurrence of sex-ratio X chromosomes in Drosophila simulans. In this species, because of the co-occurrence of drive suppressors, the natural populations and the derived laboratory strains show an equal sex-ratio even when sex-ratio X chromosomes are present at a high frequency. The presence of sex-ratio X chromosomes is established via crosses with a standard strain that is devoid of drive suppressors. In this article, we show first that the sex-ratio trait in D. simulans results from the action of several X-linked loci. Second we describe drive suppressors on each major autosome as well as on the Y chromosome. The Y-linked factors suppress the drive partially whereas the autosomal suppression can be complete. PMID- 9335601 TI - Hierarchical analysis of genetic structure in native fire ant populations: results from three classes of molecular markers. AB - We describe genetic structure at various scales in native populations of the fire ant Solenopsis invicta using two classes of nuclear markers, allozymes and microsatellites, and markers of the mitochondrial genome. Strong structure was found at the nest level in both the monogyne (single queen) and polygyne (multiple queen) social forms using allozymes. Weak but significant microgeographic structure was detected above the nest level in polygyne populations but not in monogyne populations using both classes of nuclear markers. Pronounced mitochondrial DNA (mtDNA) differentiation was evident also at this level in the polygyne form only. These microgeographic patterns are expected because polygyny in ants is associated with restricted local gene flow due mainly to limited vagility of queens. Weak but significant nuclear differentiation was detected between sympatric social forms, and strong mtDNA differentiation also was found at this level. Thus, queens of each form seem unable to establish themselves in nests of the alternate type, and some degree of assortative mating by form may exist as well. Strong differentiation was found between the two study regions using all three sets of markers. Phylogeographic analyses of the mtDNA suggest that recent limitations on gene flow rather than longstanding barriers to dispersal are responsible for this large-scale structure. PMID- 9335603 TI - Heterochromatic trans-inactivation of Drosophila white transgenes. AB - Position effect variegation of most Drosophila melanogaster genes, including the white eye pigment gene is recessive. We find that this is not always the case for white transgenes. Three examples are described in which a lesion causing variegation is capable of silencing the white transgene on the paired homologue (trans-inactivation). These examples include two different transgene constructs inserted at three distinct genomic locations. The lesions that cause variegation of white minimally disrupt the linear order of genes on the chromosomes, permitting close homologous pairing. At one of these sites, trans-inactivation has also been extended to include a vital gene in the vicinity of the white transgene insertion. These findings suggest that many Drosophila genes, in many positions in the genome, can sense the heterochromatic state of a paired homologue. PMID- 9335602 TI - Enhancement of overgrowth by gene interactions in lethal(2)giant discs imaginal discs from Drosophila melanogaster. AB - Recessive lethal mutations of the lethal(2)giant discs (l(2)gd) and lethal(2)fat (l(2)ft) loci of Drosophila melanogaster cause imaginal disc hyperplasia during a prolonged larval stage. Imaginal discs from l(2)ft l(2)gd or Gl(2)gd double homozygotes show more extensive overgrowth than in either single homozygote, and double homozygous l(2)ft l(2)gd mitotic clones in adult flies show much more overgrowth than is seen in clones homozygous for either l(2)gd or l(2)ft alone. dachsous (ds) also acts as an enhancer of l(2)gd, producing dramatically overgrown discs and causing failure to pupariate in double homozygotes. The comb gap (cg) mutation, which also interacts with ds, greatly enhances the tendency of imaginal discs from l(2)gd larvae to duplicate as they overgrow. If l(2)gd homozygotes are made heterozygous for l(2)ft, then several discs duplicate, indicating that l(2)ft acts an a dominant enhancer of l(2)gd. l(2)ft also acts as a dominant enhancer of l(2)gd, and conversely l(2)gd acts as a dominant modifier of l(2)ft. The enhancement of overgrowth caused by various mutant combinations is accompanied by changes in expression of Decapentaplegic and Wingless. These results show that tumor suppressor genes act in combination to control cell proliferation, and that tissue hyperplasia can be associated with ectopic expression of genes involved in pattern formation. PMID- 9335604 TI - Mobile element 297 in the Abd-B gene of Drosophila melanogaster, not Delta 88, is responsible for the tuh-3 mutation. AB - The tumorous-head-3 (tuh-3) mutation has been associated with the insertion of mobile element Delta 88 at +200 on the bithorax complex (BX-C) DNA map, 5' of all Abdominal-B (Abd-B) transcripts. Different phenotypes of tuh-3 are regulated by the tumorous-head-1 (tuh-1) maternal effect locus. In the presence of the recessive tuh-1h maternal effect, tuh-3 offspring produce homeotic abdominal and genital tissue in the head. In the presence of the dominant tuh-1g maternal effect, tuh-3 offspring have normal heads but now show genital defects. One other mutant, I127B, produces flies with identical defects to that of tuh-3 in the presence of both maternal effects. Molecular analysis of I127B revealed the insertion of mobile element 297 in the Abd-B gene, approximately 25 kb downstream of the Delta 88 insertion in tuh-3. No other abnormalities were detected. Reexamination of our tuh-3 strain revealed a 297 insertion in an identical region to that of I127B, in addition to the Delta 88 insertion. Recombinants of tuh-3, carrying 297 only, produced homeotic head defects and genital defects in the presence of the tuh-1h and tuh-1g maternal effects, respectively. Recombinants of tuh-3, carrying Delta 88 only, failed to produce any defects in the presence of either maternal effect. Based upon these results, we propose that it is the 297 insertion in the Abd-B gene, not Delta 88, that is responsible for the tuh-3 mutation. PMID- 9335605 TI - Homology requirements for targeting heterologous sequences during P-induced gap repair in Drosophila melanogaster. AB - The effect of homology on gene targeting was studied in the context of P-element induced double-strand breaks at the white locus of Drosophila melanogaster. Double-strand breaks were made by excision of P-w(hd), a P-element insertion in the white gene. A nested set of repair templates was generated that contained the 8 kilobase (kb) yellow gene embedded within varying amounts of white gene sequence. Repair with unlimited homology was also analyzed. Files were scored phenotypically for conversion of the yellow gene to the white locus. Targeting of the yellow gene was abolished when all of the 3' homology was removed. Increases in template homology up to 51 base pairs (bp) did not significantly promote targeting. Maximum conversion was observed with a construct containing 493 bp of homology, without a significant increase in frequency when homology extended to the tips of the chromosome. These results demonstrate that the homology requirements for targeting a large heterologous insertion are quite different than those for a point mutation. Furthermore, heterologous insertions strongly affect the homology requirements for the conversion of distal point mutations. Several aberrant conversion tracts, which arose from templates that contained reduced homology, also were examined and characterized. PMID- 9335606 TI - A genetic and mosaic analysis of a locus involved in the anesthesia response of Drosophila melanogaster. AB - We describe a genetic and behavioral analysis of several alleles of har38, a mutant with altered sensitivity to the general anesthetic halothane. We obtained a P-element-induced allele of har38 and generated several excision alleles by remobilizing the P element. The mutants narrow abdomen (na) and har85 are confirmed to be allelic to har 38. Besides a decreased sensitivity to halothane, all mutant alleles of this locus cause a characteristic walking behavior in the absence of anesthetics. We have quantified this behavior using a geotaxis apparatus. Responses of the mutant alleles to different inhalational anesthetics were tested. The results strongly favor a multipathway model for the onset of anesthesia. Mosaic flies were tested for their response to halothane and checked for their abnormal walking behavior. The analysis suggests that both the behaviors are exhibited only by such mosaics as have the entire head of mutant origin. It is likely that this focus represents an element of a common pathway in the anesthetic response to several inhalational anesthetics but not all. This result is the first demonstration of regional specificity in the CNS of any animal for general anesthetic action. PMID- 9335607 TI - Molecular population genetics of Drosophila immune system genes. AB - A striking aspect of many vertebrate immune system is the exceptionally high level of polymorphism they harbor. A convincing case can be made that this polymorphism is driven by the diversity of pathogens that face selective pressures to evade attack by the host immune system. Different organisms accomplish a defense against diverse pathogens through mechanisms that differ widely in their requirements for specific recognition. It has recently been shown that innate defense mechanisms, which use proteins with broad-spectrum bactericidal properties, are common to both primitive and advanced organisms. In this study we characterize DNA sequence variation in six pathogen defense genes of Drosophila melanogaster and D. mauritiana, including Andropin; cecropin genes CecA1, CecA2, CecB, and CecC; and Diptericin. The necessity for protection against diverse pathogens, which themselves may evolve resistance to insect defenses, motivates a population-level analysis. Estimates of variation levels show that the genes are not exceptionally polymorphic, but Andropin and Diptericin have patterns of variation that differ significantly from neutrality. Patterns of interpopulation and interspecific differentiation also reveal differences among the genes in evolutionary forces. PMID- 9335608 TI - Splice-junction elements and intronic sequences regulate alternative splicing of the Drosophila myosin heavy chain gene transcript. AB - The Drosophila muscle myosin heavy chain (Mhc) gene primary transcript contains five alternatively spliced exon groups (exon 3, 7, 9, 11 and 15), each of which contains two to five mutually exclusive members. Individual muscles typically select a specific alternative exon from each group for incorporation into the processed message. We report here on the cis-regulatory mechanisms that direct the processing of alternative exons in Mhc exon 11 in individual muscles using transgenic reporter constructs, RT-PCR and directed mutagenesis. The 6.0-kilobase exon 11 domain is sufficient to direct the correct processing of exon 11 alternatives, demonstrating that the alternative splicing cis-regulatory elements are local to Mhc exon 11. Mutational analysis of Mhc exon 11 reveals that the alternative exon nonconsensus 5'-splice donors are essential for alternative splicing regulation in general, but do not specify alternative exons for inclusion in individual muscles. Rather, we show, through exon substitutions and deletion analyses, that a 360-nucleotide intronic domain precisely directs the normal processing of one exon, Mhc exon 11e, in the indirect flight muscle. These and other data indicate that alternative exons are regulated in appropriate muscles through interactions between intronic alternative splice-specificity elements, nonconsensus exon 11 splice donors and, likely, novel exon-specific alternative splicing factors. PMID- 9335609 TI - Mismatch repair by efficient nick-directed, and less efficient mismatch-specific, mechanisms in homologous recombination intermediates in Chinese hamster ovary cells. AB - Repair of single-base mismatches formed in recombination intermediates in vivo was investigated in Chinese hamster ovary cells. Extrachromosomal recombination was stimulated by double-strand breaks (DSBs) introduced into regions of shared homology in pairs of plasmid substrates heteroallelic at 11 phenotypically silent mutations. Recombination was expected to occur primarily by single-strand annealing, yielding predicted heteroduplex DNA (hDNA) regions with three to nine mismatches. Product spectra were consistent with hDNA only occurring between DSBs. Nicks were predicted on opposite strands flanking hDNA at positions corresponding to original DSB sites. Most products had continuous marker patterns, and observed conversion gradients closely matched predicted gradients for repair initiated at nicks, consistent with an efficient nick-directed, excision-based mismatch repair system. Discontinuous patterns, seen in approximately 10% of products, and deviations from predicted gradients provided evidence for less efficient mismatch-specific repair, including G-A-->G-C specific repair that may reflect processing by a homologue of Escherichia coli MutY. Mismatch repair was > 80% efficient, which is higher than seen previously with covalently closed, artificial hDNA substrates. Products were found in which all mismatches were repaired in a single tract initiated from one or the other nick. We also observed products resulting from two tracts of intermediate length initiated from two nicks. PMID- 9335610 TI - Inheritance and mapping of Compact (Cmpt), a new mutation causing hypermuscularity in mice. AB - During selection for protein content in mice at the Technical University of Berlin, individuals showing high protein content and a compact exterior were noted. Animals showing this "Compact" phenotype were separated to form a new line. The present investigations were carried out on a Hungarian subpopulation of this line, selected for maximum expression of the Compact phenotype, and apparently at fixation for the relevant genes. Fertility and viability of the Compact subpopulation was normal. As compared to normal mice, carcass percentage values for male and female Compact mice were 9.4 and 6.8% greater, respectively; and the muscle:bone weight ratio in males was 1.61-fold greater. The Compact phenotype showed variable expressivity and was of intermediate dominance in males, but almost fully recessive in females. The hypothesis that a single gene is solely responsible for the Compact phenotype was rejected by maximum likelihood analysis. Linkage mapping using selective DNA pooling located a single locus (denoted Cmpt) strongly associated with the Compact phenotype on mouse chromosome 1. Fine mapping, using individual selective genotyping and haplotype analysis, located Cmpt to the region between D1Mit375 and D1Mit21, approximately one third of the way to D1Mit21. PMID- 9335611 TI - Developmental quantitative genetics, conditional epigenetic variability and growth in mice. AB - Ontogenetic variation in the causal components of phenotypic variability and covariability is described for body weight and tail length in mice derived from a full 7 x 7 diallel cross. Age-related changes in additive, dominance, sex-linked and maternal variance and covariance between 14 and 70 days of age are described. Age-specific variance components at time t are conditioned on the causal genetic effects at time (t - 1). This procedure demonstrates the generation of significant episodes of new genetic variation arising at specific intervals during ontogeny. These episodes of new genetic variation are placed in the context of epigenetic models in developmental quantitative genetics. These results are also concordant on recent findings on age-specific gene expression in mouse growth as shown by QTL analyses. PMID- 9335612 TI - Two dominant mutations in the mouse fused gene are the result of transposon insertions. AB - The mouse Fused locus encodes a protein that has been implicated in the regulation of embryonic axis formation. The protein, which has been named Axin to distinguish it from the product of the unrelated Drosophila melanogaster gene fused, contains regions of similarity to the RGS (regulators of G-protein signaling) family of proteins as well as to dishevelled, a protein that acts downstream of Wingless in D. melanogaster. Loss-of-function mutations at Fused lead to lethality between days 8 and 10 of gestation. Three dominant mutations result in a kinked tail in heterozygotes. Two of the dominant mutations, Fused and Knobbly, result from insertion of intracisternal A particle retrotransposons into the gene. The insertion in Fused, within the sixth intron, creates a gene that produces wild-type transcripts as well as mutant transcripts that initiate at both the authentic promoter and the 3'-most long terminal repeat of the insertion. Knobbly, an insertion of the retrotransposon into exon 7, precludes the production of wild-type protein. Thus the Fused homozygote is viable whereas Knobbly is a recessive embryonic lethal. In both mutants the dominant kink-tailed phenotype is likely to result from the synthesis of similar amino-terminal fragments of Axin protein that would contain the RGS domain, but lack the dishevelled domain. PMID- 9335613 TI - Murine albino-deletion complex: high-resolution microsatellite map and genetically anchored YAC framework map. AB - The murine albino-deletion complex developed as part of the Oak Ridge specific locus test covers 6-11 cM of chromosome 7. This complex has proven to be a valuable resource for localizing traits to a small target region for positional cloning. In this study, we mapped the endpoints of deletions in this complex using all of the available Mit simple-sequence length polymorphism (SSLP) markers. Concurrently, this mapping has determined the map order of nearly all of the SSLP markers, most of which were previously unresolved. The SSLP-based deletion map was confirmed and genetic distances were determined using the European Collaborative Interspecific Backcross panel of nearly a thousand mice. The average SSLP marker resolution is 0.3-0.4 cM, comparable to the cloning capacity of yeast artificial chromosomes (YACs). The SSLP markers were then used to construct a genetically anchored YAC framework map that further confirms the deletion map. We find that the largest deleted region distal to Tyr is about two to three times larger than the largest proximal deletion region, and the original C3H/101 regions flanking the deletions (moved to an St2A cch/cch background) are smaller than anticipated, which we suggest may result from increased recombination rates immediately flanking the deleted regions. PMID- 9335615 TI - Generating autotetraploid sporophytes and their use in analyzing mutations affecting gametophyte development in the fern Ceratopteris. AB - The haploid gametophytes of the fern Ceratopteris richardii are autotrophic and develop independently of the diploid sporophyte plant. While haploid genetics is useful for screening and characterizing mutations affecting gametophyte development in Ceratopteris, it is difficult to assess whether a gametophytic mutation is dominant or recessive or to determine allelism by complementation analysis in a haploid organism. This report describes how apospory can be used to produce genetically marked polyploid sporophytes whose gametophyte progeny are heterozygous for mutations affecting sex determination in the gametophyte and a known recessive mutation affecting the phenotype of both the gametophyte and sporophyte. The segregation ratios of wild-type to mutant phenotypes in the gametophyte progeny of polyploid sporophyte plants indicate that all of the mutations examined are recessive. The presence of many multivalents and few univalents in meiotic chromosome preparations of spore mother cells confirm that the sporophyte plants assayed are polyploid. The DNA content of the sperm of their progeny gametophytes was also found to be approximately twice that of sperm from wild-type haploid gametophytes. PMID- 9335614 TI - Detailed comparative mapping of cereal chromosome regions corresponding to the Ph1 locus in wheat. AB - Detailed physical mapping of markers from rice chromosome 9, and from syntenous (at the genetic level) regions of other cereal genomes, has resulted in rice yeast artificial chromosome (YAC) contigs spanning parts of rice 9. This physical mapping, together with comparative genetic mapping, has demonstrated that synteny has been largely maintained between the genomes of several cereals at the level of contiged YACs. Markers located in one region of rice chromosome 9 encompassed by the YAC contigs have exhibited restriction fragment length polymorphism (RFLP) using deletion lines for the Ph1 locus. This has allowed demarcation of the region of rice chromosome 9 syntenous with the ph1b and ph1c deletions in wheat chromosome 5B. A group of probes located in wheat homoeologous group 5 and barley chromosome 5H, however, have synteny with rice chromosomes other than 9. This suggests that the usefulness of comparative trait analysis and of the rice genome as a tool to facilitate gene isolation will differ from one region to the next, and implies that the rice genome is more ancestral in structure than those of the Triticeae. PMID- 9335616 TI - Analysis of recombination sites within the maize waxy locus. AB - Genetic fine structure analysis of the maize wx locus has determined that the ratio of genetic to physical distance within wx was one to two orders of magnitude higher than the average for the maize genome. Similar results have been found at other maize loci. In this study, we examined several mechanisms that could account for this pattern. First, crossovers in two other maize genes resolve preferentially at specific sites. By mapping exchanges between wx-B1 and wx-I relative to a polymorphic SstI site, we found no evidence for such a hotspot at wx. Second, deletion of promoter sequences from wx alleles had little effect on recombination frequencies, in contrast to results in yeast where promoter sequences are important for initiating recombination in some genes. Third, high levels of insertion polymorphism may suppress intergenic recombination. However, the presence of a 2-kb Ds element 470 bp upstream of the wx transcription start site did not further suppress recombination between Ds insertions in nearby wx sequences. Thus, none of these mechanisms is sufficient to explain the difference between intergenic and intragenic recombination rates at wx. PMID- 9335617 TI - The Ac-st2 element of maize exhibits a positive dosage effect and epigenetic regulation. AB - A novel derivative of the maize transposable element Ac, termed Ac-st2, that displays a positive dosage effect in maize has been identified. Although identical in sequence to other Ac elements, increasing the copy number of the element in the endosperm results in earlier and more frequent Ds excision. Ac-st2 autonomously transposes and catalyzes somatic excision of Ds elements. Germinal transpositions of either Ac-st2 or Ds, however, were not observed. The Ac-st2 phenotype includes a reduction in Ac transcript accumulation that is associated with increased methylation at specific sites in the promoter region of the major transcriptional start site within Ac (ORFa). This element differs from metastable (cycling) Ac derivatives in that Ac-st2 conditions a uniform transposition pattern throughout endosperm and plant development. Ac-st2 undergoes frequent increases in activity after its association with an active Ac element. This change in activity correlates with reduced levels of methylation in the ORFa promoter region. Using a competitive PCR assay, Ac transcript accumulation was followed through endosperm development. From these data, a model is proposed to explain the patterns of variegation associated with both "wild type" active Ac and Ac-st2 elements. PMID- 9335618 TI - Evolutionary dynamics of sporophytic self-incompatibility alleles in plants. AB - The stationary frequency distribution and allelic dynamics in finite populations are analyzed through stochastic simulations in three models of single-locus, multi-allelic sporophytic self-incompatibility. The models differ in the dominance relationships among alleles. In one model, alleles act codominantly in both pollen and style (SSIcod), in the second, alleles form a dominance hierarchy in pollen and style (SSIdom). In the third model, alleles interact codominantly in the style and form a dominance hierarchy in the pollen (SSIdomcod). The SSIcod model behaves similarly to the model of gametophytic self-incompatibility, but the selection intensity is stronger. With dominance, dominant alleles invade the population more easily than recessive alleles and have a lower frequency at equilibrium. In the SSIdom model, recessive alleles have both a higher allele frequency and higher expected life span. In the SSIdomcod model, however, loss due to drift occurs more easily for pollen-recessive than for pollen-dominant alleles, and therefore, dominant alleles have a higher expected life span than the more recessive alleles. The process of allelic turnover in the SSIdomcod and SSIdom models is closely approximated by a random walk on a dominance ladder. Implications of the results for experimental studies of sporophytic self incompatibility in natural populations are discussed. PMID- 9335619 TI - The nuclear gene Rf3 affects the expression of the mitochondrial chimeric sequence R implicated in S-type male sterility in maize. AB - The mitochondrial genomes of maize plants exhibiting S-type cytoplasmic male sterility (cms-S) contain a repeated DNA region designated R. This region was found to be rearranged in the mitochondria of all cms-S cytoplasmically revertant fertile plants in all nuclear backgrounds analyzed. A 1.6-kb mRNA transcribed from the R region in mitochondria of sterile plants was absent from all cytoplasmic revertants examined. The nuclear gene Rf3, which suppresses the cms-S phenotype, was found to have a specific effect on the expression of the R sequence; the abundance of the major R transcripts, including the cms-S-specific 1.6-kb mRNA, is decreased in mitochondria of restored plants. Nucleotide sequence analysis of R has revealed similarities to the R1 plasmid found in some South American maize races with RU cytoplasm, to the M1 plasmid found in one source of Zea luxurians teosinte, to the atp9 mitochondrial gene and its 3' flanking sequence, and also to a region 3' to the orf221 gene. The derived amino acid sequence of the R region predicts two open reading frames (ORFs). These ORFs contain the similarities to R1, M1, atp9 and orf221. The present report reveals the chimeric nature of the R region, describes the complex effect of Rf3 on the expression of the R sequence and implicates R in the sterile phenotype of cms-S maize. PMID- 9335621 TI - The evolution of recombination: removing the limits to natural selection. AB - One of the oldest hypotheses for the advantage of recombination is that recombination allows beneficial mutations that arise in different individuals to be placed together on the same chromosome. Unless recombination occurs, one of the beneficial alleles is doomed to extinction, slowing the rate at which adaptive mutations are incorporated within a population. We model the effects of a modifier of recombination on the fixation probability of beneficial mutations when beneficial alleles are segregating at other loci. We find that modifier alleles that increase recombination do increase the fixation probability of beneficial mutants and subsequently hitchhike along as the mutants rise in frequency. The strength of selection favoring a modifier that increases recombination is proportional to lambda(2)S delta r/r when linkage is tight and lambda(2)S3 delta r/N when linkage is loose, where lambda is the beneficial mutation rate per genome per generation throughout a population of size N, S is the average mutant effect, r is the average recombination rate, and delta r is the amount that recombination is modified. We conclude that selection for recombination will be substantial only if there is tight linkage within the genome or if many loci are subject to directional selection as during periods of rapid evolutionary change. PMID- 9335620 TI - An interspecific backcross of Lycopersicon esculentum x L. hirsutum: linkage analysis and a QTL study of sexual compatibility factors and floral traits. AB - A BC1 population of the self-compatible tomato Lycopersicon esculentum and its wild self-incompatible relative L. hirsutum f. typicum was used for restriction fragment length polymorphism linkage analysis and quantitative trait loci (QTL) mapping of reproductive behavior and floral traits. The self-incompatibility locus, S, on chromosome 1 harbored the only QTL for self-incompatibility indicating that the transition to self-compatibility in the lineage leading to the cultivated tomato was primarily the result of mutations at the S locus. Moreover, the major QTL controlling unilateral incongruity also mapped to the S locus, supporting the hypothesis that self-incompatibility and unilateral incongruity are not independent mechanisms. The mating behavior of near-isogenic lines carrying the L. hirsutum allele for the S locus on chromosome 1 in an otherwise L. esculentum background support these conclusions. The S locus region of chromosome 1 also harbors most major QTL for several floral traits important to pollination biology (e.g., number and size of flowers), suggesting a gene complex controlling both genetic and morphological mechanisms of reproduction control. Similar associations in other flowering plants suggest that such complex may have been conserved since early periods of plant evolution or else reflect a convergent evolutionary process. PMID- 9335622 TI - Bottleneck effect on evolutionary rate in the nearly neutral mutation model. AB - Variances of evolutionary rates among lineages in some proteins are larger than those expected from simple Poisson processes. This phenomenon is called overdispersion of the molecular clock. If population size N is constant, the overdispersion is observed only in a limited range of 2N sigma under the nearly neutral mutation model, where sigma represents the standard deviation of selection coefficients of new mutants. In this paper, we investigated effects of changing population size on the evolutionary rate by computer simulations assuming the nearly neutral mutation model. The size was changed cyclically between two numbers, N1 and N2 (N1 > N2), in the simulations. The overdispersion is observed if 2N2 sigma is less than two and the state of reduced size (bottleneck state) continues for more than approximately 0.1/u generations, where u is the mutation rate. The overdispersion results mainly because the average fitnesses of only a portion of populations go down when the population size is reduced and only in these populations subsequent advantageous substitutions occur after the population size becomes large. Since the fitness reduction after the bottleneck is stochastic, acceleration of the evolutionary rate does not necessarily occur uniformly among loci. From these results, we argue that the nearly neutral mutation model is a candidate mechanism to explain the overdispersed molecular clock. PMID- 9335623 TI - Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. AB - The main purpose of this article is to present several new statistical tests of neutrality of mutations against a class of alternative models, under which DNA polymorphisms tend to exhibit excesses of rare alleles or young mutations. Another purpose is to study the powers of existing and newly developed tests and to examine the detailed pattern of polymorphisms under population growth, genetic hitchhiking and background selection. It is found that the polymorphic patterns in a DNA sample under logistic population growth and genetic hitchhiking are very similar and that one of the newly developed tests, Fs, is considerably more powerful than existing tests for rejecting the hypothesis of neutrality of mutations. Background selection gives rise to quite different polymorphic patterns than does logistic population growth or genetic hitchhiking, although all of them show excesses of rare alleles or young mutations. We show that Fu and Li's tests are among the most powerful tests against background selection. Implications of these results are discussed. PMID- 9335624 TI - The effect of marker heterozygosity on the power to detect linkage disequilibrium. AB - The relationship between marker heterozygosity and the power to detect linkage disequilibrium is examined through the analysis of an example and through a simulation study. The analysis suggests that, despite the penalties for multiple testing incurred with multiple alleles, greater heterozygosity results in greater power. The results of the simulation study are in accord with those of the analysis. PMID- 9335625 TI - On the fertility effects of pericentric inversions. PMID- 9335626 TI - Differences between generalists and specialists. PMID- 9335627 TI - The feasibility and validity of studies comparing orthopedists and non orthopedists caring for musculoskeletal injuries: results of a pilot study. AB - OBJECTIVE: To review care provided by orthopedists and non-orthopedists for a variety of common acute injuries to the shoulder, knee, and ankle; and to evaluate the difficulties in assessing results from diagnostic data collected for administrative reporting by comparing these data with data collected by medical record abstraction for the same patients. METHODS: The Rochester Epidemiology Project database was used to identify new cases of Colles fracture, knee sprain, ankle sprain, and shoulder sprain/rotator cuff injury among Olmsted county residents > 18 years of age, who were evaluated between January 1, 1990 and December 31, 1992. Data were collected from an administrative database and medical records. RESULTS: Although the diagnoses recorded in the administrative database and those obtained from medical record review were the same in 94% of the 500 cases reviewed, only 270 (67.7%) actually represented acute injuries. Medical record review showed that (for the shoulder and ankle) injury severity was highly correlated with treatment specialty: 13 of 15 severe injuries were cared for by orthopedists, while 64 of 68 mild injuries were treated by non orthopedists. Diagnostic uncertainty was more common among non-orthopedists; in 21 of the 149 patients treated by non-orthopedists (15.1%), the final (i.e., recorded for coding) diagnosis was different from the initial diagnosis. In 14 of these cases, an orthopedist made the final diagnosis. None of the diagnoses made by orthopedists were revised. CONCLUSION: Current diagnostic coding does not capture injury acuity, severity, or diagnostic uncertainty. Studies that limit themselves to such data for diagnostic information are likely to suffer from biases which can skew results in unpredictable ways. Conclusions drawn from such studies are likely to be flawed. PMID- 9335628 TI - Satisfaction of patients attending an arthritis clinic in a county teaching hospital. AB - OBJECTIVE: To gather information about the satisfaction of medically indigent arthritis patients with their health care. METHODS: Patients attending a university-affiliated county hospital arthritis clinic were surveyed using a questionnaire about their satisfaction with various aspects of the clinic. RESULTS: Two hundred thirty-two out of 283 questionnaires were completed. Patients were most satisfied with the care given by doctors, and least satisfied with the waiting times; accessibility, environment, and information received intermediate responses. Although most patients said that they were satisfied with their overall care, only 53% would continue to attend the clinic if they had full insurance. CONCLUSION: A number of aspects of health care delivery were sources of dissatisfaction for medically indigent arthritis patients. Attention to these concerns could increase overall satisfaction and perhaps improve compliance in this group of patients. PMID- 9335629 TI - Self-efficacy for arthritis pain: relationship to perception of thermal laboratory pain stimuli. AB - OBJECTIVE: To examine how self-efficacy for arthritis pain relates to the perception of controlled laboratory pain stimuli. METHODS: Forty patients with osteoarthritis completed self-report measures of self-efficacy for arthritis pain. They then participated in a single experimental session in which measures of thermal pain threshold and tolerance were collected, as well as measures of the perceived intensity and unpleasantness of a range of thermal pain stimuli. RESULTS: Correlational analyses revealed that patients reporting high self efficacy for arthritis pain rated the thermal pain stimuli as less unpleasant than those reporting low self-efficacy. When subjects scoring very high and very low in self-efficacy were compared, it was found that subjects scoring high on self-efficacy for arthritis pain had significantly higher pain thresholds and pain tolerance than those scoring low on self-efficacy. CONCLUSIONS: These results indicate that self-efficacy for arthritis pain is related to judgments of thermal pain stimuli. Implications for the understanding of arthritis pain and for future laboratory research are discussed. PMID- 9335630 TI - The experience of rheumatoid arthritis pain and fatigue: examining momentary reports and correlates over one week. AB - OBJECTIVE: To evaluate the daily experience of patients with rheumatoid arthritis (RA) in an ecologically valid manner; Ecological Momentary Assessment (EMA) was employed. Diurnal cycles and within-day variation of self-reported pain and fatigue were examined as were relationships between pain, fatigue, daily stressful events, and sleep. METHODS: Thirty-five patients with RA were alerted with an electronic beep 7 times per day for 7 consecutive days. Assessments were recorded at each beep. Upon awakening each day, sleep information was reported. RESULTS: There were large individual differences in variation of pain and fatigue. Stressors were associated with increased pain but not fatigue. Subjects with poor sleep had higher levels of pain and fatigue. Diurnal cycles of pain and fatigue were found, yet were observed for only some patients (37% and 34%, respectively). CONCLUSION: The use of EMA deepens our understanding of the pain and fatigue experienced by RA patients. This method may help identify subgroups of patients who are highly "psychoreactive" to environmental stimuli and/or who have diurnal patterns to their symptoms. It may also be used to improve existing instruments. PMID- 9335631 TI - Measurement of depression in Mexican patients with rheumatoid arthritis: validity of the Beck Depression Inventory. AB - OBJECTIVE: To validate a Spanish version of the Beck Depression Inventory (BDI) in Mexican patients with rheumatoid arthritis (RA). METHODS: Thirty-five patients with RA seen in our outpatient clinic were included. A semistructured psychiatric interview was applied, and the following instruments were administered: the BDI, the Hospital Anxiety and Depression Scale (HAD), and the Health Assessment Questionnaire Disability Index. Diagnostic properties of the BDI for both full length and smaller versions taking out somatic items were compared against a gold standard. The gold standard for comparison was the diagnosis of depression according to the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised Criteria. RESULTS: Thirty-seven percent of RA patients had a diagnosis related to depression, most of which were major depression or dysthymia. The original BDI showed a high sensitivity (92%) and a high correlation with the HAD (r = 0.83). Exclusion of somatic items in modified versions of the BDI had a similar performance. CONCLUSIONS: The original BDI is a suitable instrument to detect depression in Mexican RA patients. Nevertheless, shorter versions without some of the somatic items also show an adequate performance. PMID- 9335632 TI - Inflammatory myopathies: issues in diagnosis and management. AB - The techniques of magnetic resonance imaging and spectroscopy have been shown to have utility in the diagnosis and management of inflammatory muscle diseases. But perhaps more important have been the new insights into the pathophysiology of these diseases which MR studies, along with new immunologic data on autoantibodies and cellular infiltrates, have afforded. Pathologic subsets of inflammatory muscle disorders have been identified, suggesting, for example, that PM and DM are distinct disorders, thereby challenging the idea that these are relatively homogeneous syndromes. Further insights into disease pathogenesis which are likely to emerge from these new findings may allow identification of etiologic factors and improved approaches to treatment. PMID- 9335633 TI - A consumer's guide to meta-analysis. AB - The same proponent of meta-analysis who feels that it offers the highest level of evidence of treatment efficacy has also warned that "it is easy to do a meta analysis; it is hard to do one well" (1). This paper has listed a series of issues that should be addressed in the construction of a good meta-analysis. A meta-analysis protocol needs to be prepared. A concerted effort should be made to capture all published and unpublished studies that address the research question. These studies should then be compared to the inclusion and exclusion criteria from the protocol to determine which will be used in the meta-analysis. A random effects analysis should be performed on the studies. A test of the homogeneity of studies should be done. Funnel plots and the tolerance for null results should be computed to assess publication biases. Finally, sensitivity analyses should be performed. All of these issues need to be discussed in the report of the meta analysis so that readers can judge the validity of the conclusions. PMID- 9335634 TI - Managed care for whom? PMID- 9335635 TI - The patient first, please. PMID- 9335637 TI - The best is yet to come. PMID- 9335636 TI - Balancing ethics and economics. PMID- 9335638 TI - Managed care, bureaucracy, and quality. PMID- 9335639 TI - Managed care through better communication. PMID- 9335640 TI - Observations on "final act". PMID- 9335641 TI - Physical restraints in the acute care setting. PMID- 9335642 TI - The re-engineering of respiratory care. PMID- 9335643 TI - Sentinel event review, Part I: A new spirit of inquiry. PMID- 9335648 TI - Telemedicine: threats and opportunities. PMID- 9335644 TI - The collective good. PMID- 9335649 TI - Awards on the brink. PMID- 9335650 TI - Oxygen therapy. PMID- 9335651 TI - Research needed into the transfer of elderly people. PMID- 9335652 TI - Clinical effectiveness and the pace of change. PMID- 9335653 TI - Infection control and related issues in intravascular therapy. AB - The use of Intravascular (i.v.) catheters has become an increasingly common practice in today's healthcare setting. This increase, however, brings associated problems. An overview of the current literature surrounding i.v. devices and infection control issues was undertaken. This included: the pathogenesis of infection with reference to local and systemic complications; the importance of handwashing; the influence of catheter material and type on infection rates; the controversy over skin preparation and whether it is best to use povidone-iodine, chlorhexidine or an antibiotic ointment; insertion techniques; the duration of line placement; the dilemma over the choice of dressing, i.e. gauze vs transparent; nurses' responsibilities in relation to the UKCC standards and professional accountability; safety issues; and the main principles surrounding the safe disposal of sharps. To conclude, this overview finds that all healthcare professionals must be held accountable for every aspect of i.v. therapy, including the control of infection. PMID- 9335654 TI - Identification of coping strategies used by heart transplant recipients. AB - Heart transplantation confronts the patient with major physical, psychological and social demands. Psychological adjustment to these stressors requires effective coping abilities. The purpose of this study was to investigate the coping mechanisms used by heart transplant recipients. A group of 42 heart transplant recipients completed the questionnaire. The instrument, F-COPES (Family Crisis Oriented Personal Evaluation Scales), was used to measure coping behaviours, thereby producing scores on five coping subscales: Acquiring social support; Reframing; Seeking spiritual support; Mobilizing family to accept help; and Passive appraisal. The results indicated that subjects in the present study (Scottish patients) scored slightly higher than normative subjects (American patients) on the Reframing subscale, slightly lower on the two subscales Acquiring social support and Mobilizing family to accept help, and much lower on Spiritual support. These results also indicated that the coping strategies used by this sample appeared to be largely independent of the time elapsed since transplantation, and the age of the subject. The major finding was that subjects scored much higher on the Passive appraisal coping subscale (almost double the score of the normative subjects). PMID- 9335655 TI - The use of restraint on patients in Israeli psychiatric hospitals. AB - This first-ever study of the use of restraint on psychiatric patients in Israel sought to determine the dominant motivations in the decision to use restraint, which patients were most likely to be restrained, and whether there was a consistent policy on the matter. A survey of the official records of every instance of restraint during one month in the closed wards of all government psychiatric hospitals, supplemented with interviews, revealed that 14.2% of the study population had undergone restraint. The declared reasons were conventional, i.e. violence, disturbed behaviour, etc. Undeclared was an interaction between patients and staff, both the most professional and the less skilled, which surprised the authors and requires more investigation. Some subgroups, e.g. women and certain immigrant groups, were restrained markedly more frequently than other groups. No consistency of policy was found. Overall, much of the restraint applied is deemed unnecessary and recommendations are made for its reduction. PMID- 9335656 TI - Towards anti-oppressive practice in mental health nursing. AB - Working in Partnership, the Department of Health's report on the 1994 review of mental health nursing, implies that mental health nurses should develop anti oppressive approaches to nursing practice. There is a notable absence of articles within the nursing literature which specifically address this issue. This is possibly because the historical and ideological issues which have informed the development of mental health nursing are complex and difficult to unravel. However, an integration of the theories of David Cooper and Frantz Fanon may provide an appropriate starting point for the development of a theory of anti oppressive practice which addresses some of the issues specific to mental health nursing. PMID- 9335657 TI - Understanding motivation to enhance patient compliance. AB - Motivation as a psychological concept is important in health care because frequently good care is dependent on the motivation of the patient, his/her family and the practitioners within the healthcare team. In addition, nurses need to be motivated to keep up to date with professional developments. This article describes George Kelly's framework of motivation and suggests recommendations to enhance practice in this area. The main conclusion is that although external determinants can encourage a person to engage in a particular activity the most effective kind of motivation is that which is internally mediated. PMID- 9335658 TI - The legal and ethical implications of consent to nursing procedures. AB - Nurses are increasingly expanding their practice to include many more invasive procedures. Consequently, there is a need to re-examine nurses' responsibilities in relation to obtaining consent for nursing as opposed to medical procedures. Fully informed consent is not a legal requirement in England, for either medical or nursing procedures. However, this article argues that to comply with the standard set by the Code of Professional Conduct nurses should obtain informed consent for any proposed procedure they undertake. The concept of informed consent is examined and applied to practice. Ultimately, nurses are charged with four key tasks in relation to securing consent for nursing procedures: educating themselves about the risks and benefits of the procedures they propose to undertake; conveying this information to patients; assessing their understanding of the information given; and endeavouring to support the patient in his/her decision. PMID- 9335659 TI - Legal problems in the operating theatre: learning from mistakes. AB - Nurses and other professionals have a legal as well as a professional duty to keep up to date with developments and changes in their clinical field of practice. Healthcare professionals should also be aware of major studies and the complaint and litigation trends in their specialties. This information can inform practice and lead to improvements in the quality of patient care. The information can often be found in the healthcare literature and the general press. This article looks at examples of the reported cases and research on negligence in theatres and discusses the legal and professional duty to keep up to date. PMID- 9335661 TI - Is euthanasia a 'quick-fix' to the dying process? PMID- 9335660 TI - Should euthanasia be legalized? PMID- 9335662 TI - A comparison of tools for the assessment of sleep pattern disturbance in critically ill adults. AB - Sleep pattern disturbance is a common nursing diagnosis among critically ill adults. Using nursing judgement in selecting the sleep assessment tool to be used in a critical care unit is a complex process that can help identify the insomnia more precisely and can lead to nursing interventions tailored to the individual patient's needs and situation. PMID- 9335663 TI - Postoperative factors contributing to prolonged length of stay in cardiac surgery patients. AB - In order to provide high quality, cost effective care, nurses need to identify and address factors that prolong the length of stay in various patient populations. The authors identify the postoperative factors contributing to prolonged length of stay in a cardiac surgery patient population and recommend collaborative management strategies to address these factors. PMID- 9335664 TI - Pathophysiology of fever. Part 2: Relooking at cooling interventions. AB - Presently, fever is thought to play a beneficial role in the control and containment of infection by curtailing virus replication, enhancing natural killer cell activity, and killing of natural killer-resistant leukemia cells. The first article in this set of articles on fever physiology, which was printed in a previous issue, covered the role of cytokines in fever. This article expands the physiology to describe the body's positive response to fever, so critical care nurses can decide which patients to cool, which ones not to cool, and how to revise cooling protocols. PMID- 9335665 TI - Making clinical decisions using SvO2 in PICU patients. AB - The use of continuous SvO2 monitoring in the critically ill pediatric patient helps guide the critical care nurse to make clinical decisions that individualize care and improve patient outcome. This article examines SvO2 and its components, oxygen delivery and oxygen consumption, as they apply to the pediatric intensive care patient. PMID- 9335666 TI - Developing an orientation program for assistive personnel in the ICU. AB - Assistive personnel are being used in intensive care unit (ICU) settings in response to the demand for more cost-effective nursing care. Nurse managers, educators, and advanced practice nurses must address the unique orientation needs of these workers. A comprehensive, competency-based orientation program for assistive personnel can help the ICU technician be successful. Despite the multitude of assistive personnel being used in the critical care settings, little information is available regarding the development of an orientation program for ICU technicians. This article discusses one ICU's experience in developing an orientation program for their ICU technicians. PMID- 9335668 TI - Inaugural lecture. Nursing and the counter culture for cancer. AB - The discourse surrounding cancer is that of warfare and of a militaristic endeavour aimed at helping those with the disease survive. The consequences of such a discourse have not previously been explored in detail; in particular the culture of the 'cancer clinic' has not been examined for how a battle mentality might effect the nature of care and treatment. This paper explores how the biomedical response to cancer and its treatment has established approaches which neglect the more day to day experiences of people living with the disease, and the suffering, disability and distress that it causes. A radical reconstruction of the culture surrounding cancer is called for, so that a new environment of care might be offered. This should be more participatory, collaborative and empowering of people with cancer and their families. It should also shift the dominant theme in cancer management from 'survival' to the here and now, and to the needs and problems people have. An argument is made for the contribution that nursing might make to such a reconstruction through developing new approaches to symptom and problem management, reorganising cancer services so that they are more supportive and patient focused, and through nursing research studies aimed at radically changing the process of research itself. PMID- 9335667 TI - Editing a nursing practice book. AB - Publishing a nursing practice book can be an exciting challenge for the critical care nurse who identifies a gap in the literature. This article provides information on how to produce an edited book from the starting point to successful publication. Identifying a book idea, selecting chapter authors, finding a publisher, writing a prospectus, the editing process, production, and marketing the book are discussed. Examples drawn from the authors' experiences in editing a transplantation nursing book are provided, as well as comments from other book editors. PMID- 9335670 TI - Swedish nurses' estimation of fatigue as a symptom in cancer patients--report of a questionnaire. AB - Many studies show that chronic fatigue is the most frequently reported symptom related to cancer and its treatment. In order to evaluate the problem in Sweden, a questionnaire was mailed to 442 registered nurses in the autumn of 1995 with the aim of determining cancer nurses' views of the nature and causes of cancer related fatigue and which, if any, nursing interventions they employed in the management of this problem. The response rate was 49%. The responses showed that these nurses regarded fatigue as the most common symptom in cancer patients, but there were few established nursing interventions. Also, nurses wanted further education and tools for evaluation of fatigue, its causes and treatment. PMID- 9335669 TI - Total parenteral nutrition for patients receiving antineoplastic therapy at a regional oncology unit: a two-year study. AB - The aim of this prospective study was to establish the exact role parenteral nutrition has in the provision of nutritional support to patients receiving antineoplastic therapy. The diagnosis, reasons for implementation, method of delivery and duration of nutritional support were determined. The outcome of nutritional support was established by the percentage change in weight and alteration in body mass index during the period of nutritional support. The results indicate that parenteral nutrition can successfully maintain the body weight of patients who are unable to receive enteral nutrition whilst receiving antineoplastic treatment. However, it is recommended that the provision of parenteral nutrition is evaluated at regular intervals and alternative feeding methods via the enteral feeding route are accessed as soon as possible. PMID- 9335671 TI - Cancer beliefs, attitudes and preventive behaviours of nurses working in the community. AB - A study of nurses, comprising district, school and practice nurses and health visitors, working in the community was carried out in 1992/1993. The aims of the research included exploration of the nurses' beliefs about and attitudes to cancer and their own cancer preventive behaviours. Focus group methodology was used, with 11 discussion groups, totalling 86 nurses. Data were analysed qualitatively and reported in relation to emergent themes. The nurses found cancer a terrifying disease, dreading a personal diagnosis and feeling ambivalent about treatment effectiveness. However, they were good role models for cancer prevention, eating a healthy diet, protecting themselves from the sun, attending regularly for cervical and breast screening, and the majority did not smoke. PMID- 9335672 TI - The management of xerostomia: a review. AB - Xerostomia causes a great deal of morbidity in patients with advanced cancer. However, it is an area which has received little attention. Indeed current management of xerostomia in palliative care units is based largely on anecdotal evidence. There are a number of studies in progress specifically looking at patients with advanced cancer, but until these are published we should take note of studies done in other patient groups. This article reviews the medical literature on the symptomatic management of xerostomia as a whole, with particular reference to treatments that are currently available in the United Kingdom. PMID- 9335673 TI - Evaluation of the introduction of the Groshong central venous catheter into the oncology/haematology department of a district general hospital. AB - Technological advances in the medical management of oncological and haematological malignancies have seen the introduction of long-term central venous access as common practice in this setting. The result of this has been an influx of a variety of lines onto the market, thus creating a minefield of choice for the practitioner. As a newly appointed Macmillan clinical nurse specialist for oncology and haematology to a unit with no established policy regarding the use of central venous catheters, the author was presented with an opportunity to introduce the most advanced line into clinical practice. In this centre the decision was made to introduce this line for a number of reasons, not only for the benefits associated with enhanced patient safety but also because of the educational and supportive role Bard Access Limited had offered. When instigating change and development it was of paramount importance that all members of the health care team were well educated regarding the use and maintenance of these lines to ensure success. This paper aims to share our experience of the Groshong line and includes a detailed audit of each line placed in a 12-month period. The information collated includes details regarding insertion method, infection rates, episodes of thrombosis and numbers of catheters removed due to malfunction. PMID- 9335674 TI - To randomise or not to randomise: methodological pitfalls of the RCT design in psychosocial intervention studies. AB - We are seeing evidence of more studies investigating the effectiveness of psychosocial interventions of cancer patients, predominantly within groups. As roles within cancer and palliative care diversify, specially trained nurses and other health care workers are taking a more active role within psychosocial intervention studies. Frequently, these studies are randomised controlled trials (RCTs). Often, the results of these psychosocial RCTs have been laid open to general criticisms of design, implementation and reporting. The following paper focuses specifically on the general and experimenter problems in conducting RCTs within psychosocial interventions. It highlights the limitations and inherent problems seen with RCTs of psychosocial interventions so that health care workers are aware of these before considering undertaking psychosocial RCTs with cancer patients. PMID- 9335675 TI - Patient satisfaction with conscious sedation for ambulatory colonoscopy in a community hospital. AB - Patient acceptance of ambulatory colonoscopy is important for colon cancer screening programs to be successful. The goal of this study was to assess patient satisfaction with the method of conscious sedation that is standard in most endoscopy units. The authors also attempted to identify patient characteristics associated with a poor sedation experience. The findings suggest that 90% of ambulatory patients undergoing colonoscopy are adequately sedated with the standard meperidine midazolam regimen. The only patient characteristic found to predict poor sedation response was the preceding use of a prescription narcotic as an outpatient. PMID- 9335676 TI - Nasogastric tube feeding and medication administration: a survey of nursing practices. AB - Using a 62-item, investigator-developed mailed questionnaire, this descriptive study of 350 randomly selected staff nurses sought to identify variations in practices in the care of patients with nasogastric tubes (NGT). Reported here are the results from the 35 questionnaire items related to NGT feedings and medication administration. Wide variations were found in the amount of gastric residual considered "excessive," as well as in whether the entire residual was returned to the stomach. A large percentage of the nurses relied on physicians' orders for gavage feeding rate, giving additional water, and using the liquid form of a medication. Data indicate that published nursing research is not consistently used in practice, which represents a theory-practice gap. In addition, a theory-practice deficit in several areas related to NGT feedings points to the need for further research. PMID- 9335677 TI - Basics of constipation. AB - In this article, the author defines and describes the causes of constipation. Physiology of the colon is reviewed, and the nurse's role in helping patients and families with constipation is also described. PMID- 9335678 TI - The nurse who is the patient: a personal perspective. AB - The author, a registered nurse, describes her personal experience living with a chronic illness. The emotional turmoil she experienced as a patient with ulcerative colitis and later as a patient with a J-pouch heightened her awareness of the importance of learning self-management skills and the necessity of emotional support for all patients with a chronic illness. Implications for nurses are discussed throughout the article. PMID- 9335679 TI - Methane gas explosion during colonoscopy. AB - This article discusses a complication of colonoscopy that healthcare providers think can not happen: methane gas explosion. Despite proper bowel cleansing, such an incident did occur. This article shows how quick recognition and action can save a patient's life. Colonic preparations are also discussed. PMID- 9335681 TI - Position statement: role delineation of the licensed practical/vocational nurse in gastroenterology and/or endoscopy. PMID- 9335680 TI - Position statement: role delineation of the registered nurse in a staff position in gastroenterology and/or endoscopy. PMID- 9335684 TI - Caring for interpersonal violence survivors. AB - Interpersonal violence is a serious public health problem in the United States. Millions of violent injuries occur each year, but the magnitude of the problem can only be estimated. The National Child Abuse and Neglect Data System reported that 3,140,000 abused or neglected children were reported in 1994 to child protective services agencies. PMID- 9335685 TI - Do I need a flu shot? AB - Home care nurses soon will be confronted with questions about influenza prevention. Studies show that high-risk patients who should take the vaccine are more likely to do so if they understand its efficacy and absence of side effects. PMID- 9335683 TI - Assisted suicide: the wrong answer. AB - One of the most complex and troubling issues facing society and health care professionals is physician-assisted suicide (PAS). The convergence of significant trends in legislation, judicial decisions, research, and public sentiment has highlighted its importance. A broad spectrum of societal opinion and philosophical, religious, legal, and professional debate surrounds PAS. PMID- 9335686 TI - Home-based immunoglobulin therapy reduces incidence of neonatal RSV infection. AB - Respiratory syncytial virus (RSV) infections are responsible for more than 90,000 hospitalizations and 4500 infant and childhood deaths each year. RSV infection, which peaks between January and March, can cause respiratory problems, such as bronchiolitis and pneumonia, in children younger than 2 years old. PMID- 9335687 TI - Understanding viral hepatitis B. AB - Epidemic jaundice, known today as viral hepatitis, was described by Hippocrates nearly 2500 years ago. Yet only in the past 20 years or so have the main viruses that cause hepatitis been elucidated. During this short interval, the main modes of transmission and effective methods of prevention also have been described. PMID- 9335688 TI - Congestive heart failure readmission. AB - Multiple hospital readmissions average between 21% and 27% in the United States yearly. Little is understood about what causes this rate and, more importantly, how readmission can be prevented. An integrative review examined 13 research articles in an attempt to identify specific factors that lead to readmission. Risk factors continually researched were dependency, patient age, stage of illness, hospital length of stay (LOS), prior hospitalization, care after discharge, and mobility status. Congestive heart failure (CHF) and chronic obstructive pulmonary disease (COPD) were the medical conditions responsible for most readmissions. No single factor was found to universally predict readmission, but several were statistically significant. Because of the results of this review, a study was conducted to determine whether clients with a primary or secondary diagnosis of CHF who receive home health care and a systematic education program would be less likely to be readmitted to the hospital or have a shorter LOS if readmitted than home health clients who did not receive the education program. PMID- 9335689 TI - More than a best friend. PMID- 9335690 TI - A family systems approach to case management in a rural community setting. AB - Case management has proven to be an effective mechanism in assisting clients to seek and receive health care in the most appropriate setting. A family systems approach has been added to the case management program in a Midwestern rural community to encourage greater client and family participation in developing solutions to health care problems. Clients and families become more autonomous decision-makers and authors of change. PMID- 9335691 TI - LabScam: could you be involved? PMID- 9335693 TI - Is a deemed status option through the Joint Commission right for your home health agency. PMID- 9335692 TI - Wound care 2000: challenges to integration across the continuum. PMID- 9335694 TI - Who are home medical equipment services providers? PMID- 9335695 TI - Oral cancer screening in the elderly: the home healthcare nurse's role. AB - Screening for oral cancer is an often overlooked element of annual medical examinations and physical assessments in the elderly. As the elderly population increases, more emphasis will be placed on caring for clients in the home, and oral cancer screening will be provided by nurses. This article outlines the most common types of oral cancer and provides step-by-step instructions of performing an oral cancer screening examination. PMID- 9335696 TI - The occupational therapist as a member of the home healthcare team. AB - In the changing healthcare environment of the 1990s, home healthcare professionals are being challenged to provide successful, cost-effective, and appropriate skilled care for patients. It is important that all professional home healthcare personnel work as a team to accomplish these goals. The OT is an important member of this team. After an acute illness or throughout the course of a chronic illness, there are numerous times when home healthcare patients benefit from skilled OT services. Home healthcare nurses are in a unique position to recognize the need to work with the patient's physician for referral to other members of the healthcare team. PMID- 9335697 TI - Our patient wanted to stay home, but... PMID- 9335698 TI - Community assessment for the home healthcare nurse. AB - Home health clients and their families live within the context of a community. To deliver effective care, home health nurses must understand how the community influences the health of their clients and families. This article describes a community assessment, how to incorporate it into daily practice, and suggests strategies for using the data to enhance the quality and efficiency of home healthcare. PMID- 9335699 TI - Community health nursing: can being self-employed work for you in home care? AB - There is a fine distinction between being an independent contractor and being an employee. The advantages of being self-employed as a community health nurse are many. Self-employment suits new parents, graduate students, people in transition, with more than one profession, and who don't want a fixed schedule. However, this type of nursing is not for everyone. A broker such as CHN can help nurses become successfully self-employed. At a time when hospitals are downsizing and home care is becoming more in demand, brokers such as CHN provide a framework in which busy, experienced, community health nurses can work when and where they want. Good clinical and communication skills and a wish to be autonomous are necessities. A willingness to travel to different agencies and a reliable car are also important. A love for variety, flexibility, and independence make self employment as a home health nurse a clinician's dream. PMID- 9335700 TI - The geropsychiatric clinical nurse specialist in home care. AB - Home care nurses often are asked to provide care to older adults with multiple problems, including psychiatric diagnoses. This article identifies areas of knowledge a general nurse needs to provide care for geropsychiatric patients and describes the role of the geropsychiatric clinical nurse specialist as part of a home care team. PMID- 9335702 TI - The future of ASORN. American Society of Opthalmic Registered Nurses. PMID- 9335703 TI - Education, training, and experience: the professional triad that nurses bring to the ophthalmic health care table. AB - As health care continues its rapid and seemingly unending helix of change, I am uncomfortably mindful that I must be ever vigilant to convince the many evaluators of health care services that ophthalmic registered nurses are a vital and unique part of the team. I continually ask myself the question: what is it about nursing that is so unique and necessary to health care, whether it is at the bedside, in the community, operating theater, or clinic? And what is it that ophthalmic nurses bring to the "table" that other providers of ophthalmic health care services don't or at least, should not? When I look again at this second question, the should not really grabs me. Ophthalmology battles optometry over scope of practice issues and takes them all the way to the courts of law in order to obtain backup. It's time for ophthalmic nurses to stand for what they believe is their rightful scope, even if we do not have the power or money to take it to our judicial system. PMID- 9335704 TI - Patient satisfaction survey of an outpatient eye clinic. AB - The patient population seen at the Veterans Affairs Health Care System is unique because of the chronic health problems that are prevalent in the elderly. In addition to ocular diseases, arthritis, hypertension, hearing impairments, and heart disease, mental health problems are shown to affect 15% to 25% of the elderly. Consequently, elderly patients often present with complex problems that are difficult for both staff and patients. This, in addition to other problems, such as late arrival for appointment as a result of impaired mobility, multiple symptoms and concerns, multiple medications, and complicated medical history makes the clinic appointment challenging for all. A patient satisfaction survey was initiated in the Seattle Veterans Affairs Eye Clinic to try to identify specific problems and look for solutions to improve quality and efficiency of care for our nation's veterans. PMID- 9335705 TI - Vision and vision loss. PMID- 9335706 TI - Lymphoma. AB - Lymphomas are rare tumors, which can be unilateral or bilateral. Lymphomas may affect the conjunctiva, the eyelid, and the orbit; and they may be diagnosed systemically. Ophthalmic nurses are the perfect health care professionals to educate and provide the care for these patients. An interesting patient who was diagnosed with a lymphoma is presented. PMID- 9335707 TI - The care and feeding of a fledgling chapter. AB - The success of an organization is determined by the dedication and commitment of its members. This essay aptly illustrates how members of the American Society of Ophthalmic Registered Nurses (ASORN) assisted in the formation of the 34th local chapter of ASORN and improved the strength and professionalism of registered nurses working in ophthalmology. PMID- 9335708 TI - The value of the ORN. Ophthalmic Registered Nurse. AB - In my last message I said I would meet again with Dr. Dunbar Hoskins, Executive Vice President of the American Academy of Ophthalmology (AAO) to provide further support of the distinct role of the ophthalmic registered nurse (ORN) on the ophthalmic health care team in the private practice or clinic setting. The value of the ORN is well established in the operating room and at the bedside; however, the efficacy of the ORN is questioned routinely in the office in light of today's. health care climate of bottom line costs. What follows are excerpts of my discussion with Dr. Hoskins, based in part on my own clinical experiences, but also on the many scenarios, examples, and sketches provided me by countless ASORN members. When I "tapped" you for help, you responded most powerfully. Thank you all. PMID- 9335709 TI - The mentor commitment. AB - An active mentoring program can benefit an entire organization. The nurses being mentored always have a resource for questions and support. Orientation, inservice, and continuing education can be individualized, based on the needs of the nurse and the areas of work. Continuing education, so essential for professional development, can become an everyday occurrence, as it is an integral part of mentoring. Other benefits are those reaped by the mentors through the recognition and affirmation of their expertise. Difficult problems often require simple solutions. Using the expert ophthalmic nurse to mentor the novice can be one of these solutions. Through a mentor connection, job satisfaction, leadership, and professional empowerment can be a reality. In addition, achieving this will help us face a future in which the ophthalmic nurses' role will be at the forefront. PMID- 9335710 TI - Pupillary examination: do i really need to look? AB - A pupillary examination takes very little time and does not require expensive equipment. Unfortunately, many ophthalmic personnel do not take the time to thoroughly evaluate pupillary function. Pupillary examinations can be easy if divided into the specific elements to be assessed. These elements should include color, shape, size, equality, and reaction to stimulus. This article will briefly review some of the more common pupillary abnormalities and review the steps of the pupillary examination. PMID- 9335711 TI - Retinopathy of prematurity. AB - Research studies have provided advancements in knowledge regarding retinopathy of prematurity. Examination identifies premature infants with threshold disease that indicates need for treatment. It is hoped that timely evaluation and treatment will reduce the incidence of ROP-related blindiness. PMID- 9335712 TI - Visual field screening for glaucoma. PMID- 9335713 TI - Multistate Regulation Task Force explores mutual recognition model for nursing regulation. PMID- 9335714 TI - Georgia Board of Nursing discovers the power of positive regulation. PMID- 9335715 TI - Oklahoma Nursing Education Program approval process explained. PMID- 9335716 TI - Utah Board of Nursing approves schools of nursing. PMID- 9335717 TI - U.S. Secretary of Education and board of nursing recognition: why and how? PMID- 9335718 TI - Telecommunications and health care. PMID- 9335719 TI - Teagle LPN to BSN Initiative: a success story. AB - Success was ultimately reflected by the lack of difference in the performance of the generic BSN nursing student and the LPN to BSN student. While there were slight variations in NCLEX-RN examination pass rates-LPNs, 100 percent; control group of generic students, 97 percent-this initiative showed the enormous potential of a predominantly female population of health care providers who have frequently been discounted by baccalaureate nursing education. Some of the critical factors leading to success in the Teagle Initiative seem to be: an interested, supportive faculty, counseling support, curricula revised to acknowledge the LPNs' prior learning, and tuition and resource support. These success stories prove that, by nursing doing the unexpected and continuing to open its educational arms to those in the health care provider family who require additional knowledge, skills and abilities, the health care environment benefits. PMID- 9335720 TI - National Council studies continued competence. Committee develops Personal Accountability Profile. PMID- 9335721 TI - Medicare managed care and the role of care management. PMID- 9335722 TI - What's funny about case management? AB - Humor is an effective tool to prevent and resolve burnout, a common problem associated with the practice of case management. Easily accessed by almost everyone, humor has many physiological and psychological benefits. The conscious use of humor in an organization requires a commitment at all levels in order for the organization to benefit as a whole. Offering formal seminars, using humor in company newsletters, and encouraging cartoon or joke sharing impacts on the physical and mental health of the employees. Techniques for bringing laughter to the workplace are outlined, as well as examples from the practice of case management. PMID- 9335724 TI - Preparing health care organizations for successful case management programs. AB - This article reports the results of a study of four hospital-based providers in varying stages of implementing case management programs. Three of the providers had most of the necessary elements in place to ensure success, such as a mix of reimbursement sources, an effective and integrated information management system, a full range of clinical services, and continuous quality improvement programs. The authors make several suggestions for key activities that must be pursued by any health care organization seeking to implement a case management program in an era of managed care, tightening reimbursement, and consumer demand for quality care. These include the need to (a) organize essential case management functions under a centralized structure; (b) set realistic, quantifiable targets, and (c) design a communications plan for the program. PMID- 9335725 TI - Delivering home-based case management to families with children with mental retardation and developmental disabilities. AB - To meet the needs of individuals with mental retardation and developmental disabilities (MR/DD) and their families living in urban setting, a noncenter based model of case management was implemented. In contrast to traditional case management in which families and consumers come to the case manager and most service coordination is done by telephone or in meetings at the case manager/social worker's worksite, the case manager in a noncenter-based model is mobile and able to meet the consumer and family in their domains. In this model, case management is provided in conjunction with in-home residential habilitation and funded by Medicaid under the Home and Community Based Services Waiver. This funding stream provides monies for nontraditional services delivered in noncertified settings. Case managers used the Family Resource Scale to get an immediate indication of the resources and needs of each family. The scale highlights the adequacy of a person's basic and caregiving resources, as well as financial needs. The findings from this study suggest that an understanding of both disability and entitlements is essential for case managers who may have to help advocate for consumers around services and benefits. Moreover, to build and maintain an egalitarian and supportive relationship with families, the importance of caregiver-specified resources and needs must be recognized by case managers. Access to resource information and the ability to engage the family in problem solving depends on a well-trained staff with the ability to respond to individuals with different needs and from a variety of circumstances. These essential skills prepare a case manager to assist families with their immediate requirements as well as to mobilize them to plan for future needs. PMID- 9335723 TI - Peer intervention in case management practice. AB - Using peers as case managers in dealing with current and chronic public health problems such as substance abuse, gang violence, or the HIV/AIDS crisis has been shown to improve outreach efforts, monitoring, and outcomes in hard-to-reach populations. This article focuses on a case management strategy that uses peer modeling interventions to assist people in renegotiating their present life circumstances. Peer modeling engages peers of the client population as case managers and employs group-mediated, social control intervention strategies in the community to bring about positive changes in lifestyle and living conditions. The peer approach is an enhanced version of case management, utilizing the core activities of outreach, assessment, planning, linking, monitoring, and advocacy but adding peer-led, skill-based training activities, coupled with a system of positive incentives designed to encourage a more healthful lifestyle. To clarify this enhanced approach to case management, the authors present a matrix to illustrate how key case management activities might be enhanced through peer modeling interventions. We conclude by suggesting the circumstances in which an organization responsible for service delivery might consider using peer modeling in addressing difficult public health problems, and we discuss the advantages and disadvantages of such a strategy. PMID- 9335726 TI - Unrecognized elder abuse victims. Older abused women. AB - Many older women are being abused by the family members they love. Spouses, partners, or adult children can use physical, sexual, or emotional abuse as tactics to dominate and control their victims. Current responses by case managers do not address older abused women's needs for safety and support. Case managers who believe caregiver stress is the primary cause of elder abuse often do not assess domestic violence in later life. Services offered in cases of caregiver stress are significantly different than those recommended for domestic abuse. Offering inappropriate interventions can put older abused women in greater danger. This article will give an overview of elder abuse and specifically domestic abuse in later life. An empowerment model for working with victims, strategies for working with the abuser, and suggestions for developing a coordinated community response are also discussed. PMID- 9335727 TI - Case management and Mexican-American elders. AB - This article describes four levels of assimilation of elderly Mexican-Americans that may assist social workers to deliver culturally sensitive case management services. This delivery can be better achieved if social workers recognize the client's level of assimilation and develop case management plans based on this information. Descriptions of the prototype and its use are provided. The prototype can also be used to sensitize social workers to subtle differences that exist in many clients who identify themselves as being members of a minority group. PMID- 9335728 TI - Ophthalmology clinic cross training: a case study. PMID- 9335729 TI - The diagnosis and management of the dry eye. PMID- 9335730 TI - Traumatic cataract and Wieger's ligament. PMID- 9335731 TI - Prevalence of uveitis in an outpatient juvenile arthritis clinic: onset of uveitis more than a decade after onset of arthritis. PMID- 9335733 TI - Triage for the contact lens patient. PMID- 9335732 TI - Treatment of primary intraocular cancers: retinoblastoma and uveal malignant melanoma. PMID- 9335734 TI - Self-assessment quiz: dermatomyositis. PMID- 9335735 TI - Likely outcomes surrounding child protection and criminal record checks. PMID- 9335736 TI - Come fly with me. AB - We have both worked performing medical retrievals of tourists who have become injured or sick overseas and required a nursing escort. Back to Australia, such services are covered by travel insurance policies. Through this work we have had the opportunity to visit, albeit briefly, some beautiful places with some fantastic health care facilities (we use the term 'fantastic' in the entire spectrum of its meaning!). All our adventurous stories are true, but those recounted here are an amalgam of a number of trips and a number of patients. The places, names and diagnoses of patients have been changed to protect the hospitals, and those people in our care. PMID- 9335737 TI - Nursing in Saudi Arabia. AB - The experience of nursing in Saudi Arabia has brought mixed reactions from Australian nurses. Some are shocked by the conditions they found in the local hospitals and the restrictions on women's lives which apply to them as well. For others, Saudi offers the chance to earn a high income, travel easily and extensively and experience a unique ancient culture. Conditions can often be very different from those cultured in nursing contracts, so the principle caveat interior, buyer beware, should be your guide. Here two nurses recount their own experiences of nursing Saudi style. PMID- 9335738 TI - Lobby for The Aged Care Bill. PMID- 9335739 TI - Legal issues for nurses. PMID- 9335740 TI - Three years in the USA. PMID- 9335741 TI - Child care in Australia. PMID- 9335742 TI - The story of midwifery is part of nursing's history. PMID- 9335743 TI - Conditions for psychatric nurses critical. PMID- 9335744 TI - Men are victims too. PMID- 9335746 TI - War memorial to Australian Services nurses. PMID- 9335745 TI - Health and safety update. PMID- 9335747 TI - Nurse in profile. Nives Houlihan. PMID- 9335748 TI - Mental health. The aboriginal experience. PMID- 9335749 TI - Getting your share of the health dollar: maintaining community health funding. PMID- 9335750 TI - Keep the nursing in nursing homes campaign. PMID- 9335751 TI - Community nursing information system update. PMID- 9335753 TI - An ongoing challenge. Providing palliative care nursing in the community. PMID- 9335752 TI - Post acute community care. Western Sydney Area Health Service (WSAHS). PMID- 9335754 TI - Feet are for life. An interdisciplinary approach to foot and leg ulcer management. PMID- 9335755 TI - Nursing the net: searching for information. PMID- 9335756 TI - NSW Nurses' Association guidelines for the provision of basic foot care by nurses. PMID- 9335758 TI - The Workers Compensation legislation. PMID- 9335757 TI - The Australian health care system. PMID- 9335759 TI - Sleeping well: the art of shiftwork. PMID- 9335760 TI - Diabetes: the role of the enrolled nurse in managing a diabetic patient. PMID- 9335762 TI - Nurse in profile. Gail Mackenzie. PMID- 9335761 TI - Tax deductions for nurses. PMID- 9335763 TI - On the front line. Nursing and complementary therapies. AB - Wide-range of complementary therapies are being used by nurses to deal with the health issues encountered in standard nursing practice. The Lamp looks at some hands-on examples-the soothing jets of the hydrobath, the healing power of music, the pain relief given by therapeutic massage and the efficacy of essential oils. PMID- 9335764 TI - NSW Nurses' Association. Complementary therapies in nursing practice [guideline]. PMID- 9335765 TI - The Health Department position. Complementary therapies in mainstream medicine. PMID- 9335766 TI - The AMA position. Complementary therapies & other health professionals. PMID- 9335769 TI - Representing nurse managers--is there a conflict of interests? PMID- 9335767 TI - Learning to heal the body, mind and spirit. PMID- 9335770 TI - The A-Z of complementary, natural & alternative therapies. PMID- 9335771 TI - Menopause: a natural part of life. PMID- 9335772 TI - Complementary therapies is an area of practice which is of to many nurses. PMID- 9335773 TI - Fatigue and driving performance. PMID- 9335775 TI - Nurse in profile. Michele Simpson. PMID- 9335774 TI - Implementation of better practice management principles in all facets of its operations. PMID- 9335776 TI - Technician training. PMID- 9335777 TI - Improving outcomes. PMID- 9335778 TI - "Guts to glory"/Part I. Taking the idea of a new dialysis facility from bricks to shiny reality. PMID- 9335779 TI - Adding patient feedback on quality of life to the outcomes assessment picture. PMID- 9335780 TI - The ultimate gift. PMID- 9335781 TI - A look at potential ESRD treatment alternatives for the future. Xenotransplantation. PMID- 9335782 TI - Immunosuppressants of tomorrow. PMID- 9335784 TI - DOQI guidelines/fourth in a series. Adequacy, HD dose, reuse, compliance. NKF Dialysis Outcomes Quality Initiative. PMID- 9335783 TI - Love, Sissy. A sibling's view on organ donation. PMID- 9335785 TI - DOQI guidelines/third in a series/Part II. Nutrition, pediatrics, patient selection. NKF-Dialysis Outcomes Quality Initiative. PMID- 9335786 TI - How exercise can change "disability" to "ability" in the ESRD patient. PMID- 9335788 TI - Decline in nursing school enrollment continues according to latest NLN study. PMID- 9335789 TI - Program evaluator workshops prove successful in preparation on accreditation visits. PMID- 9335790 TI - Preceptorship for RGNs in a small community hospital. PMID- 9335791 TI - En career development opportunities. PMID- 9335792 TI - Palliative care--breathlessness. PMID- 9335793 TI - Open learning--interaction at work. Conscious and unconscious processes. PMID- 9335794 TI - Open learning--relationships in a climate of change. Changes in working practices. PMID- 9335795 TI - How to reference effectively. PMID- 9335797 TI - Relieving AIDS-related pain. PMID- 9335798 TI - Administering oxygen by mask. PMID- 9335799 TI - Researching health care topics on the Internet. PMID- 9335801 TI - Tips for pediatric i.v. insertion. PMID- 9335800 TI - Sexual assault in a skilled-nursing facility. PMID- 9335802 TI - Action stat--bacterial meningitis. PMID- 9335803 TI - Managing hemorrhage--taking the right steps to protect your patient. PMID- 9335804 TI - Special delivery--using a syringe pump to administer i.v. drugs. PMID- 9335805 TI - My needlestick. PMID- 9335806 TI - Assessing a heart murmur. PMID- 9335808 TI - What I learned from Jackie. PMID- 9335807 TI - Critical thinking--digging deeper for creative solutions. PMID- 9335810 TI - Just say no--positively. PMID- 9335809 TI - How to help your patient with epilepsy. PMID- 9335813 TI - Dealing with diversity in the workplace. PMID- 9335812 TI - Managing antibiotic-resistant organisms. PMID- 9335814 TI - One step forward. PMID- 9335815 TI - Lost in space? PMID- 9335816 TI - Multicultural health care: reconciling universalism and particularism. AB - One of the issues of concern in multicultural health care is the degree to which one universal (or mainstream) service can meet the needs of all groups and the extent to which specialist (or ethno-specific) services are required to meet the needs of particular groups. In order to advance debate on this issue, multiculturalism is examined against concepts of identity, equality, bureaucracy and participation. While the appeal of universal healthcare services is that they appear to deliver equal health care to all, they in fact systematically advantage those whose values most closely fit with the dominant social norms. Although ethnic-specific services may overcome this problem, in that they enable 'tailor made' care, it is unlikely that they would be able to meet all of the health needs of all people from ethnic minorities, especially in locations where numbers are low. Ethnic minority participation in the processes of the health system is proposed as a way for the universal health system to reconcile the need to treat people equally (universalism), but in accordance with their unique and different needs (particularism). PMID- 9335817 TI - Transcultural nursing and a care management partnership project. AB - This paper aims to illustrate how Leininger's Theory of Culture Care Diversity and Universality has influenced the research process of a study that emerged from a care management partnership between Canadian nursing teachers and Tunisian nurses. The purpose of the study was to investigate the meanings of care as viewed by university hospital-based Tunisian nurses. The qualitative analysis of data gathered through observation-participation and interviews highlights recurrent patterns and reveals three major professional care themes. For Tunisian nurses care means to secure the patient's cooperation towards the medical regimen within established rules in the hospital; to contribute to curing the patient by using current technology as well as by maintaining their technical skills and improving their medical knowledge; to take charge of the patient to assist the physician in treating disease. This study showed that Tunisian nurses emphasize curing rather than widely shared community values such as interdependence, intercommunication, understanding, presence and responsibility for others. Discussion of the study's findings draws upon the perspective provided by Freire's Oppressed Group Theory. In order to promote cultural congruence within the Care Management Partnership Project in Tunisia, the three predicted modes of care within Leininger's theory guide the decisions and actions for future nursing research and partnership activities. PMID- 9335818 TI - Is there anyone in there? Psychiatric nursing meets biological psychiatry. AB - Mental health nursing operates without a clear idea of the nature of both mind and mental. Increasingly, this lack of a defensible theoretical position has led to an increasing dependence on the concept of mind now current in biological psychiatry. But the materialist monism of biological psychiatry is itself open to doubt, particularly concerning its dependence on a priori assumptions about the nature of the relationship between mind and brain. The reductionism and objectivism inherent in this approach necessarily ignores that aspect of mind most germane to nursing, the first-person nature of the mental. An alternative is briefly sketched which stresses the mediation of behaviour by the brain, rather than viewing behaviour as causally related to brain processes. Adoption of this approach conserves nursing's focus on the subjective experience as being paramount. Nursing education should, therefore, be wary of incorporating the biological approach without a critical analysis of its suppositions and of the conception of human nature which it supports. PMID- 9335819 TI - Grounded theory method, Part II: Options for users of the method. AB - Era-specific issues are presented for the consideration of the potential grounded theorist. Two of the issues are general to grounded theory method, while the other four assist in clarifying aspects relevant to the selection of a specific mode of the method. Five broad options regarding multiple possible modes of grounded theory method are suggested and detailed. The inquirer is not limited to the two major modes usually presented and discussed in the literature. Users of the method are challenged to continue contributing to the development of the method, while justifying and debating methodological modifications. PMID- 9335820 TI - Life and the laundromat: reflections on dirty linen and everyday private life. AB - This is a paper in which I reflect on, and draw issues from, an unplanned ethnographic experience in a London laundromat. The concept of 'everyday life' has a kind of simple, even benign, quality. However, there are many events in everyday life that are complex and complicating. Everyday life can be mundane- boring even. But it can also confront and trouble us, even when it concerns such apparently ordinary matters like doing the laundry. This paper is about how the ordinary matters of everyday life can become problematic and how our involvement in them can confront us with dilemmas that are unwanted yet require our attention and judgement as participants in social life. PMID- 9335821 TI - Changing conceptions of practical skill and skill acquisition in nursing education. AB - The learning of practical skill in nursing is not a priority of nursing education today. This is a result of a narrow and mechanistic concept of practical skill and its acquisition in nursing education. This paper reviews these conceptions as they have been developed in major textbooks for nurse teachers and in research articles on skill teaching, learning and evaluation over the past 50 years. Nursing research must move out of the laboratory and into the clinical setting in order to incorporate the typical aspects of practical skill learning in nursing. Future challenges in this kind of research are discussed. PMID- 9335822 TI - An ethic of caring: conceptual and practical issues. AB - The purpose of this paper is to explore the concept of an ethic of care. Caring as a concept has long been associated with nursing, but in more recent times there has been a strong move to promote an ethic of care as a fundamental concept of nursing. Literature and a personal perspective are used to explore the concept of an ethic of care. It is a premise of this paper that care should be taken that an ethic of care, as currently described, does not contribute to nurses' powerlessness in ethical decision-making in the clinical area. Issues that need to be addressed include the inadequate articulation of the concept to inform and guide practice, and also the emotively laden language utilized by some writers. It is concluded that an ethic of care shows promise to advance nursing knowledge and practice and requires increased research and dialogue to clarify the concept. PMID- 9335823 TI - O, brave New World: heritability and schizophrenia. PMID- 9335824 TI - Paola di Guilio in a conversation with Margaret Dunlop. Interview by Paola di Guilio. PMID- 9335825 TI - Cultural frameworks of nursing practice: situating the self. PMID- 9335826 TI - Manuscript reviewing: too long a concealed form of scholarship. PMID- 9335827 TI - Report on 'the Virtual University?' symposium. Melbourne, Australia, 21-22 November 1996. PMID- 9335828 TI - Yes, we have no bananas... PMID- 9335829 TI - Outcome-oriented research on certification. PMID- 9335830 TI - Labor law update--Part 4. AB - Many hospitals are eliminating positions as part of the restructuring process. If these decisions involve race, nationality, age or gender discrimination, back pay and punitive damages may be awarded. Cases involving wrongful termination issues are discussed. PMID- 9335832 TI - Expediting Medicare appeals. AB - Health maintenance organizations (HMOs) must expand their appeals process for Medicare members needing care urgently. They also must now accept any physician's conclusion that the member's medical circumstances warrant expedited review, regardless of whether the physician is part of the HMO's network. HMOs must inform their Medicare members how to navigate the appeals maze. PMID- 9335831 TI - What is age-specified competence? AB - About 35% of the 1,238 hospitals surveyed in the first 9 months of 1996 received Type I recommendations regarding the provision of evidence of age-specific competence for staff. This column answers questions commonly asked about age specific competence, how it is measured and how a facility can comply with this JCAHO requirement. PMID- 9335833 TI - CIOs and trends in health care computing. AB - What are the chief information officers thinking? The 1997 Health-care Information and Management Systems Society/Hewlett Packard Leadership Survey gives us a glimpse. The study's results involving information technology present some ambitious plans. PMID- 9335834 TI - Managing fraud and abuse risks. Subacute care: creating alternatives. AB - Fraud and abuse risks in subacute care concern three major areas: (1) inadequate treatment and quality of care; (2) underutilization of services; and (3) misrepresentation of service capabilities. Strategies on how to manage these risks and guide changes are given. PMID- 9335835 TI - A patient classification system for ambulatory care. AB - Patient classification systems (PCSs) for ambulatory care are difficult to develop because there are so few similarities among groups of patients and/or diagnoses in relation to the required nursing activities. In addition, the nursing activities required for specific patients and/or diagnoses also are dissimilar in relation to nursing time required. Therefore, a PCS was developed to assess staffing needs. This outpatient classification system reflects actual nursing time spent with each patient. PMID- 9335836 TI - Fusing roles--the ambulatory care nurse as case manager. AB - Most pathways concentrate on reducing the patient's length of stay. One multidisciplinary pathway team aims for a broader approach-promoting wellness as a means to avert hospitalization. Primary treatment teams provide and direct care for a group of patients, resulting in an outpatient system that is accessible and comprehensive. PMID- 9335837 TI - Reorganizing competency programs. AB - An ambulatory care educational council was formed to validate competency levels, provide staff education programs and determine individual needs. The council's goals and accomplishment are discussed. PMID- 9335838 TI - Health Care Workforce Regulation reform. AB - All health care professionals are affected by the continual changes in health care delivery and education. Nurses, in particular, are concerned that current practice, regulation and licensure issues will change the very nature of nursing as a profession. This article discusses health care regulation issues and the recommendations of the Pew Commission's Taskforce on Health Care Workforce Regulation. PMID- 9335839 TI - Moving health care education into the community. AB - The A+ Asthma Club, an educational program developed for elementary school children in inner-city schools, is offered through a series of six sessions during school hours with an additional three booster sessions. This article describes how the program was designed, its theoretical basis, the curriculum and its staffing. PMID- 9335840 TI - Developing quality assurance programs in ambulatory surgery. AB - Cost and efficiency are only two outcomes considered in evaluating surgical ambulatory services. More important, quality of care and the patient's physical and emotional health and well-being must be evaluated. Nurse practitioners play a key role in initiating and implementing quality assurance programs to meet these outcomes. PMID- 9335841 TI - Handling clinical incidents. AB - Completing a clinical incident report (CIR) when an unexpected "event" occurs is the first step in identifying practice issues and potential risk management problems. The CIR's subsequent path is explained, including the role that the peer review process plays in identifying substandard care. PMID- 9335842 TI - Fine-tuning preadmission surgical services. AB - A preadmission procedure for surgical services prepares patients both physically and psychologically. A VIP folder holds all the information patients and the surgical staff need: procedure information, test results, discharge instructions, etc. On the day of the surgery, patients report directly to the surgical unit. PMID- 9335844 TI - Developing a competency test for ambulatory care nurses. PMID- 9335843 TI - How sweet it is: glucose monitoring equipment and interpretation. AB - When monitoring blood glucose, a number of factors should be considered, e.g., the equipment and methods used to measure the levels. Teaching patients how to use this equipment is of paramount importance; outcomes depend on patient motivation and the proper use of the devices and strips. The significant advancements in glucose monitoring equipment and the lab monitoring methods are discussed. PMID- 9335846 TI - Leadership: the essentials. PMID- 9335847 TI - Have disciplines fallen? PMID- 9335848 TI - The Parse research method through music. PMID- 9335849 TI - Rogerian health patterning: evolving into the 21st century. PMID- 9335850 TI - Theoretical substruction: a guide for theory testing research. AB - This article demonstrates how theoretical substruction with its various levels of abstraction can be used as a guide for theory testing research. The article describes a method for substructing research hypotheses as well as an entire study. Substruction and its benefits, including assuring the internal consistency of model testing, are discussed as applied to a study testing hypotheses derived from proposition two of Orem's self-care deficit theory of nursing. This approach to theory testing is congruent with the coherence theory of truth and promises to contribute to the advancement of nursing epistemology. PMID- 9335852 TI - Rogerian science: opening new frontiers of nursing knowledge through its application in quantitative research. AB - This article discusses the theoretical and methodological considerations in selecting Rogers' science of unitary human beings as a framework for quantitative research. It elucidates how Rogerian science guided the research process in selected nursing studies. PMID- 9335851 TI - Contrasting two approaches in a community-based nursing practice with older adults: the medical model and Parse's nursing theory. AB - This article contrasts the assumptions and concepts of two distinct approaches, the medical model and Parse's human becoming theory, as applied in a nurse managed capitated community healthcare program for older adults. Many nurses incorporate aspects of the medical model into their practice without fully appreciating the implications or being aware of alternative perspectives. Within capitated health programs a nursing practice based on a nursing theory which emphasizes an intersubjective dialogue with older adults as they move toward different meanings and free choice is seen as more likely to be associated with greater satisfaction and reduced healthcare expenditures than a nursing approach based on the medical model, which relies on the objectification of human experience. Nurses can best balance the problem-focused orientation of a medical model dominated healthcare system by adopting an approach committed to recognizing that persons are the coauthors of their existence. PMID- 9335854 TI - Quality of life and health for persons living with leprosy. PMID- 9335853 TI - Use of Neuman's lines of defense and resistance in nursing research: conceptual and empirical considerations. AB - The generation of testable nursing theory requires operationalization of the broad concepts embedded in nursing's conceptual models or grand theories. Operationalization of the Neuman systems model concepts, the flexible line of defense and the lines of resistance, are explored in this article. Conceptual and empirical concerns imposed on the researcher when employing the model are discussed. PMID- 9335856 TI - Get a move on. Interview by Dina Leifer. PMID- 9335855 TI - Fighting for freedom to prescribe. PMID- 9335857 TI - For the record. PMID- 9335858 TI - Court cases. PMID- 9335859 TI - Special focus: health care assistants. Care assistants share our aims. PMID- 9335860 TI - Special focus: health care assistants. We must remain professionals. PMID- 9335862 TI - Mental health research network. AB - In the latest report based on information supplied from the database of the Network for Psychiatric Nursing Research in Oxford, we highlight working being carried out in a range of areas of interest to mental health nurses. This week we focus on community psychiatric nurses in primary care, research into specialisation in mental health nursing and 'revolving door' syndrome. PMID- 9335861 TI - 100 days and counting. Interview by Norah Casey. PMID- 9335863 TI - Writing a research abstract: structure, style and content. AB - The communication of ideas is fundamental to the development of all professions. Producing an abstract is an essential part of this process and it requires careful planning if it is to fulfil its purpose correctly. This article explains how to compose and use the abstract of a research paper to its full potential. PMID- 9335864 TI - Ethical dilemmas faced by mental health nurses. AB - This paper considers the dilemmas faced by mental health nurses when they encounter difficult ethical decisions. Using a case study, the author analyses the patient's right to autonomy and the nurse's duty of care. Theories of decision-making are assessed for their relevance to the work of mental health nurses. The author proposes that Hare's (1981) theory of universal prescriptivism offers an appropriate framework and examples from clinical practice are described to support this argument. PMID- 9335866 TI - Nurse education: attrition rates in the UK. AB - Student attrition is not a new phenomenon, and for many years governments have examined ways to avoid losing nurses from the profession. This article examines the problem from a historical perspective, seeks out examples of good practice and discusses their relevance to the present situation. PMID- 9335865 TI - Patient information for pain control in palliative care. AB - In this article the author argues that written information about pain and its control should be given routinely to patients and carers. This would be in line with guidelines for palliative cancer care, would relieve distress for patient and carer, and promote best practice. PMID- 9335867 TI - Psychological considerations of stoma care nursing. PMID- 9335869 TI - Dury service. PMID- 9335868 TI - Days of the Raj. Interview by Rebecca Coombes. PMID- 9335870 TI - Hard labour. PMID- 9335871 TI - Talking about a revolution. PMID- 9335872 TI - Clueless in the Commons. PMID- 9335874 TI - Lasting impressions. PMID- 9335873 TI - Homes on the range. From hell hole to heaven. PMID- 9335875 TI - Homes on the range. From hell hole to heaven. PMID- 9335876 TI - Mental health--the secret history. PMID- 9335877 TI - Interpreting policy at local level. PMID- 9335878 TI - Weed control. PMID- 9335879 TI - Providing needle exchanges in A&E. PMID- 9335880 TI - Intermittent catheterization for post-operative urine retention. AB - This article describes a small study that looked at the safety and effectiveness of intermittent catheterization in managing post-operative urine retention on a gynecological ward. PMID- 9335881 TI - Complementary therapy to fight drug addiction. PMID- 9335882 TI - Vegetarian nutrition for pregnant women. PMID- 9335883 TI - Volunteers combat social isolation in older people. PMID- 9335884 TI - Intergenerational work with older Asian people. PMID- 9335885 TI - Palliative care. Nausea and vomiting. PMID- 9335886 TI - A time for renewal. PMID- 9335887 TI - Patients, machines, and staff nurses. AB - Forty-one patients for whose direct care at least one machine was used and 33 registered nurses from the same five non-critical care units as the patients and from one related unit, participated in a semistructured tape-recorded interview to identify the core categories of the human-machine interface in clinical nursing practice and the relationships between them. Constant comparative analysis was used to organize and process the data. Patients perceived the machines as neutral because of their view of health care and because nurses were the interface between them and the machines. Nurses perceived the machines as either positive or negative, depending on their effect on the nurses' professional competence and the extent to which they worked directly with them. PMID- 9335888 TI - Just another trauma patient? PMID- 9335889 TI - Clinically based managed mental health care. AB - Managed care models are on a continuum in terms of the amount of clinical involvement of the managed care agent. Managed care constitutes a fundamental shift in provider-reimburser relations. In the literature problems with managed care are identified and examined in the context of this shifting relationship. Models at three points on the continuum of clinical involvement are reviewed with regard to the identified problems, including two models with full clinical involvement. It is predicted that clinically based models will flourish and replace models with less clinical involvement because full clinical involvement solves many problems associated with other models. PMID- 9335890 TI - Ethics of consumer rights in managed care. PMID- 9335891 TI - A nursing case management model for rural hospitals. AB - Providing health care to vulnerable rural populations presents many challenges and limitations that urban models may not address. The model of nursing case management presented here focuses on improving quality, reducing costs, and increasing access for people in rural areas through a professional case management role. The manager's role includes individual advocacy, clinical practice, education, research, and system advocacy. Empowerment of nurse case managers influences achievement of goals and job satisfaction and is based on a new view of power. The model is applicable to hospital, home, and community settings. PMID- 9335892 TI - Health locus-of-control beliefs in Vietnamese clients with latent tuberculosis. AB - The resurgence of tuberculosis constitutes a global emergency. In the United States, the number of reported cases is highest in foreign-born persons, and among that group is highest among Asians. Effective treatment requires an understanding of Asian health beliefs. Vietnamese clients were studied to identify associations between an Asian culture and the tendency to operate from an "internal" or "external" (chance or powerful others) locus of control in health beliefs. Internal-locus clients scored highest overall. Males scored significantly higher on external powerful other. Nurses are uniquely positioned to educate these internally motivated clients toward responsible health behavior and to initiate a decline in the rising rate of Asian TB in the United States. PMID- 9335893 TI - Researching nurses as educators. AB - This study uses a survey to investigate the nurse educator's perceptions of their work and goals. Respondents were educators selected from a national list of nurses with doctorates. The survey used a combination of objective and open-ended questions. The purpose was to seek a correlation between the interests of the faculty and the expectations of their hiring institutions. Suggestions are made on ways educators, clinicians, and administrators can collaborate and consult for their mutual benefit. PMID- 9335894 TI - Is the battle over? PMID- 9335895 TI - How do you help family members of patients with cancer prepare for end-of-life care at home? PMID- 9335896 TI - Cultivating the seeds of cultural competence is a growth process. PMID- 9335897 TI - "The Good Air Gang" teaches children at an early age the importance of staying smoke-free. PMID- 9335898 TI - The role of the mental health counselor in the psychiatric liaison service of the general hospital. PMID- 9335899 TI - The attitudes of physicians toward prolonging life. PMID- 9335901 TI - Notes on a contextual approach to medical ward consultation: the importance of social system mythology. PMID- 9335902 TI - Psychiatric aspects of life-threatening illness: a course for medical students. PMID- 9335900 TI - Intrusions of territory and personal space: an exploratory study of anxiety inducing factors in hospitalized patients. PMID- 9335903 TI - Etiologic factors in ulcerative colitis: birth, death and symbolic equivalents. PMID- 9335904 TI - Coronary artery bypass operation: psychological and medical problems. PMID- 9335905 TI - Ward staff problems with abortions. PMID- 9335907 TI - Psychiatric consultation with patients who refuse medical care. PMID- 9335906 TI - Psychiatric-orthopedic liaison in the hospital management of the amputee war casualty. PMID- 9335908 TI - The integration of psychosocial care in a general hospital: development of an interdisciplinary consultation program. PMID- 9335909 TI - Clinical-medical issues for the health professional who is also a leukemic. PMID- 9335910 TI - Confinement and ego regression: some consequences of enforced passivity. PMID- 9335911 TI - Psychiatric management of patients who threaten to sign out against medical advice. PMID- 9335912 TI - Neuropsychiatric complications in burn patients. PMID- 9335913 TI - Psychotherapy of a suicidal, terminal cancer patient. PMID- 9335915 TI - A portrait of Michael Balint: the development of his ideas on the use of the drug "doctor". PMID- 9335914 TI - Is psychiatric consultation in abortion obsolete? PMID- 9335916 TI - Do psychiatrists have a common language? PMID- 9335917 TI - Adapting to illness through family groups. PMID- 9335918 TI - The general practitioner and the patient with backache. PMID- 9335919 TI - Management after myocardial infarction: a controlled trial of the effect of group psychotherapy. PMID- 9335920 TI - Videotape in the training of medical students in psychiatric aspects of family medicine. PMID- 9335921 TI - Self-induced abscesses: a diagnostic and treatment enigma. PMID- 9335922 TI - Chronic pain in a victim of attempted homicide. PMID- 9335923 TI - [Hepatitis B and C virus in chronic alcoholic patients: prevalence and influence on liver injury]. AB - The aim of this prospectively designed study is to analyse the prevalence of HBV and HCV infections in 115 chronic alcohol abusers, their relation to epidemiological variables, and their meaning in pathogenesis and severity of alcoholic liver injury. A prevalence of 13.9% anti-HBc and 20.0% anti-HCV reactivity (EIA II) were found, significantly higher that found in blood donors (3.75 and 0.65% respectively). It is striking our finding of 69.6% "sporadic" type of HCV infection. Histological diagnostic of chronic hepatitis was done in 3 cases, all of them reactive to anti-HCV, enhancing the ethiologic role of HCV in the so called "alcoholic chronic hepatitis". No differences in histological final diagnosis were found related to HBV and HCV markers reactivity, suggesting no clear influence of viral infections on the severity of liver damage in alcoholics in our series. Neither anti-HCV positivity ratio seemed have to influence on these results. Despite a high prevalence of HBV and HCV infection in chronic alcohol abusers, our finding suggest no clear role for them in histological damage. PMID- 9335925 TI - [Role of inducible nitric oxide synthase in acute gastric mucosal damage in stress, in rats]. AB - In groups of female Wistar rats, someted to stress by immobilization and immersion in 18 degrees C water during 6 hrs, the role of nitric oxide (NO) in its pathophysiologic was studied. Agonist and antagonist of the isoforms Constitutive NO Synthase (cNOS) and of the inducible NO Synthase (iNOS) were used. Was found that the overdose of L-arginine aggravated of stress acute gastric lesions. The agonist of iNOS the NMDA and the antagonists dexamethasone and aminoguanidine were used. The NMDA aggravated the stress acute gastric lesions; in contrast, dexamethasone and aminoguanidine given a evident protection of the gastric mucosa in stress. Was concluded that the production of NO given by iNOS, play a role important in the pathophysiologic in stress acute gastric lesions. PMID- 9335924 TI - [Value of the emergency therapeutic endoscopy in gastrointestinal hemorrhage]. AB - The experience in therapeutic digestive endoscopy is presented using injection technique with adrenalin-polidocanol of gastrointestinal hemorrhage lesions. One hundred and twenty nine patients were treated endoscopically; the medium age was 60 years; in almost 80% of the cases, peptic ulcer disease were found. The effectiveness was 91.4% when one session procedure was used and in some cases two sessions were applied, giving a final total arrest of hemorrhage of 95.3%. The total mortality still remain high (16.3%) even without bleeding, due to coexisting poor general conditions of the patients. The technique of endoscopic injection is reviewed and the final results of our work are presented. PMID- 9335926 TI - Antegrade colonic enema (ACE): a new therapeutic approach to chronic constipation. AB - Continent appendio-cecostomy is a recently developed operation for the therapy of chronic constipation, refractory for other treatment modalities. Two patients are described in whom the operation led to a dramatic improvement of their quality of life. Technical details of the operation are offered as well as some data on the possible mechanism of colonic motility in this pathological disorder. PMID- 9335927 TI - Role of copper zinc superoxide dismutase in the short-term treatment of acetic acid-induced colitis in rats. AB - The present study describe the role of copper zinc superoxide dismutase (CuZnSOD) in the short-term treatment of experimental colitis induced with 8% acetic acid. The colonic mucosa damage index in the group of 10 rats treated intravenously with 30,000 U/kg CuZnSOD was significantly decreased when compared with the control group (10 rats) treated with normal saline (0.4 +/- 0.6 vs 1.5 +/- 0.5 p < 0.01). Assay of SOD in the control group was 0.3 +/- 0.08 and in the SOD treated group, SOD was significantly increased to 0.8 +/- 0.1, glutathione peroxidase was 44.8 +/- 6.3 in the control and 56.4 +/- 9.1 in the treated group (difference not significant). Both myeloperoxidase activity (14.0 +/- 2.5 vs 22.7 +/- 2.5) and lipid peroxidation products (13.8 +/- 2.9 vs 52.9 +/- 9.6) were significantly lower in the colonic mucosa of the SOD treated group in comparison with the control. These results indicate that the anti-inflammatory effects of CuZnSOD were mainly the removal of oxygen free radicals and indirectly the prevention of lipid peroxidation. This study suggests that CuZnSOD may be beneficial in the treatment of patients with ulcerative colitis. However, our experimental data suggest that this treatment will not have strong effects on the control of severe ulcerative colitis. PMID- 9335928 TI - [Genomic heterogeneity of hepatitis B virus, genotype A circulating in the metropolitan area of Buenos Aires, Argentina]. AB - HVB DNA was extracted from highly purified Dane particles, from sera of HBV viremic patients, collected in the metropolitan area of Buenos Aires (Argentina). HBV DNA was cloned in pUC18 vector, amplified in Escherichia coli DH5 F'. Plasmids were recovered and analyzed for HBV DNA inserts. Three recombinant plasmids, pHB4, pHB7 and pHB20 were selected, and HBVDNA inserts sequenced. The resulted sequences were incorporated at the GenBank, with the following accession numbers: PHB4P3=U33188; PHB4P5=U33189; PHB7P3=U33190; PHB7P5=U33191 and PHB20=U33190; PHB7P5=U33191 and PHB20=U33187. All belongs to the genotype A, pHB4 and pHB20 have a very close relation in between each other and with L13994 sequence, from North America origin. pHB7 have a significant distance from pHB4 and pHB20 and have a discrete homology with m57633 detected in Philippines. pHB4 shows a mutation at the T 3182-Leu in the preS1 region that change Pro for Leu, this mutation is absent in 125 sequences selected (having a 65% or more of homology) from NCBI by Blast algortm. The sequence of the pre C regions of all three inserts do not show any evidence to belong to the e-or scape mutants. Type A genotypes shows to be common in the area, but a hight degree of divergence have been demonstrate between two circulating strains. PMID- 9335929 TI - [Gastric vascular ectasia (GAVE) or watermelon stomach (WS): infrequent cause of anemia]. AB - The gastric vascular ectasia (GAVE) or watermelon stomach (WS) is an unfrequent cause of anemia or evident upper gastrointestinal bleeding in elderly patients. We presented five female patients, average age 79 years, 4 of them with a long evolution anemia and one with melena. Three of them showed a typical WS endoscopy, 2 of them with diffuse patent. All 5 cases with positive pathologic findings: vascular ectasia, fibrin thrombi and fibromuscular hyperplasia. The endoscopic biopsy is as accurate as the study of the antrectomy piece. None of them had portal hypertension although the GAVE would be different entity from the cirrhotic vascular gastropathy. The treatment consisted in monopolar electrocoagulation of the lesions after the failure with the medical treatment in one case, corticosteroid and ferrous therapies in the three cases and one of them didn't require treatment up to now. PMID- 9335930 TI - [Endoscopic treatment of pancreas divisum]. AB - The pancreas divisum is the most frequent congenital anomaly of the pancreas. It is caused by the absence of fusion in the dorsal and ventral pancreatic buds, so that most of the gland drains through the Santorini duct in the minor papilla. It has been suggested that the relative obstruction of the pancreatic pain and pancreatitis in these patients. According to this the treatment consists in facilitating the minor papilla drainage via surgery or endoscopy. Two endoscopic treatment cases are presented. In both pancreatic duct stent and further sphinterotomy were performed. This treatment and their follow-up are evaluated, coming to the conclusion that this method should be taken into account in the future, due to its viability and low morbimortality. PMID- 9335931 TI - [Lactose deficient intestinal absorption (LDIA) in a population of healthy Mexican children studied with the hydrogen ion test in air breathing]. AB - We studied 100 healthy children looking for lactose malabsortion. We performed in all of them the lactose breath test. We found a 10% with lactose malabsortion. There was no correlation between lactose breath test and fecal reducing substances. PMID- 9335932 TI - [Antibiotic-induced diarrhea: recent findings about Clostridium difficile]. AB - From the beginning of the antibiotherapy, diarrhea frequently occurred as a side effect of the treatment. The spectrum of diarrheal disease associated with antibiotic therapy ranges from antibiotic associated diarrhea and colitis, to the more severe pseudomembranous colitis, which is always associated with Clostridium difficile (CD). Because most of the antibiotics are not active against this sporulated Gram positive anaerobe. The pathogenic process occurs only by production of toxins. CD is now recognized as one of the most important hospital infections. Although therapy is available for the majority of patients with CD infection, pseudomembranous colitis is a serious disease which, if untreated, can lead to major complications. PMID- 9335933 TI - [Brazilian consensus on Helicobacter pylori and associated diseases]. PMID- 9335934 TI - [Endoscopic treatment of acute digestive hemorrhage not associated with varices]. PMID- 9335935 TI - [Gastric acid secretion inhibitors: H2 receptor antagonists (H2RA), proton pump inhibitors (PPI) or acid pump antagonists (APA)]. PMID- 9335936 TI - [New frontiers in the surgical treatment of chronic calcifying pancreatitis]. PMID- 9335937 TI - Secondary metabolites and their role in evolution. AB - The role of secondary metabolites in evolution will be examined with the view that they are chemicals released within a system by one component which have evolved to affect other component(s) within the system. Secondary metabolites are a natural outgrowth and consequence of an increase in complexity, and they are a critical part of the chemical "glue" that holds a system together. An analysis of secondary metabolites from a broad perspective (e.g. genetics, ecology, evolution, etc.) suggests that the nature of secondary metabolism can be viewed as a critical component of an emergent system (ecological) arising from a host of interlocking cycles and feedback processes. PMID- 9335938 TI - Age-related effects of diazepam on retention of inhibitory avoidance and shuttle avoidance tasks in rats. AB - We investigated the effect of diazepam on memory of 30 days-old and 60-70 days old female Wistar rats, using two behavioral tasks: step-down inhibitory avoidance (IA) and shuttle avoidance (SA). Diazepam (0.2, 1.0 or 5.0 mg/kg) or its vehicle were given i.p., 60 min prior to the training session. Training-test interval was 24 h. Diazepam impaired the retention of IA in 30 days-old rats at the three doses used, while retention of SA was not impaired by any dose. In the 60-70 days-old animals, diazepam at the dose of 0.2 mg/kg was facilitatory in IA and had no effect on SA, while doses of 1.0 mg/kg and 5.0 mg/kg impaired retention of both tasks. We suggest that these age-dependent effects of diazepam on memory of IA and SA could be related to developmental changes in brain GABAA receptors. PMID- 9335939 TI - Why Oedipus and not Christ?: a psychoanalytic inquiry into innocence, human sacrifice, and the sacred--Part I: Innocence, spirituality, and human sacrifice. PMID- 9335940 TI - Reassurance in analytic therapy. PMID- 9335941 TI - Precursors to masochistic and dependent character development. AB - Due to the extreme states of masochism, dependency, and narcissistic rage that these patients experience, the treatment must be attuned to the inevitable periods of regression and primitive defense. The patient feels compelled to be a servant to the object, yet is furious at this less than equal status. The alternating states of idealizing the object in a masochistic fashion, the anger at the lack of self importance, and the desperate hope for soothing from that object create an externally focused character structure, which generally leads to a pattern of acting out, the lack of internal linking processes, and a scarcity of interpersonal skills that foster mutuality. A fundamental lack of self soothing leads to a perpetual search and craving for the soothing talents of the object. It is often unconsciously believed that compliance and servitude will bring about this gift of soothing from the object. Hans Loewald (1962) had described internalization as a process whereby the child reaches out to take back from the environment what has been removed from him in an ever-increasing manner since his birth. For the forgotten hero, this theory is definitely true. Not only is the treatment of this type of patient one of gradual internalization of new internal object relationships and the working through of the older more pathological ones, but it is a true understanding between patient and analyst of the original nontolerable removal of the uniqueness of the self via reality and/or fantasy states. When this situation is focused upon and worked with, the taking back from the environment can occur in a spirit devoid of former states of envy, hate, resentment, and wild craving that were formally protected and disguised in a facade of dependent masochism. The patient has essentially experienced or perceived his parents, usually the mother, as not providing a soothing function or a proper fit for his developing ego. The patient has then gone about constructing various methods to compensate for this lack. The analyst is often experienced as not providing an adequate soothing function, but this emptiness is warded off from conscious expression with a compromise formation of dependent, masochistic, or narcissistic methods of relating. This style of compromise hides any envy, hunger, or rage and keeps alive the hopes of being rewarded with soothing. The patients expects to be used as a waste disposal unit and believes that this dumping by the object into him is the longed-for love. Ideally, interpretations focus upon the hunger for the soothing function of the object and the drive to obtain it at any cost. The patient will fiercely resist because he believes that in giving up his masochism and dependency, he would expose his envy and narcissistic injury. This patient believes he would then lose any hope of ever receiving the soothing function due to the fantasized destruction of the source of that soothing, his beloved object. PMID- 9335942 TI - Reflections. PMID- 9335943 TI - What postmodernism can do for psychoanalysis: a guide to the postmodern vision. PMID- 9335944 TI - What postmodernism can do for psychoanalysis: a guide to the postmodern vision. PMID- 9335945 TI - What postmodernism can do for psychoanalysis: a guide to the postmodern vision. PMID- 9335946 TI - What do all the apolipoprotein E receptors do? PMID- 9335947 TI - The fate of lipoprotein cholesterol entering the arterial wall. AB - Recent findings have helped to explain the fate of cholesterol entering the arterial wall. LDL can undergo both fusion and aggregation. These changes may cause increased retention of LDL in lesion connective tissue matrix and LDL uptake by macrophages. In the cornea, apparent fusion of LDL occurs in the absence of macrophages. Mast cells may be important in LDL fusion, as mast cell derived proteases can induce fusion of LDL through proteolysis of apolipoprotein B. LDL in arterial wall atherosclerotic lesions was found to be sialic acid-poor and ceramide-enriched. These chemical changes promote LDL aggregation. Processes that may function to remove cholesterol from the arterial wall have been reported. Macrophage-produced apolipoprotein E can mediate macrophage cholesterol efflux and macrophages can convert cholesterol to 27-oxygenated products that macrophages excrete. Alternately, another oxygenated sterol, 7-ketocholesterol, impairs macrophage cholesterol efflux. In addition, mast-cell derived chymase proteolyses HDL and reduces its capacity to stimulate cholesterol efflux. PMID- 9335949 TI - Phospholipid metabolism in cholesterol-loaded macrophages. AB - Macrophage foam cells in atherosclerotic lesions accumulate free cholesterol as well as cholesteryl ester. In addition, these cells have an increased rate of phospholipid biosynthesis and accumulate intracellular phospholipid-containing membrane structures ('whorls'). Studies with cultured macrophages have revealed the possible molecular mechanism and biological relevance of these observations. A rate-limiting enzyme of phosphatidylcholine biosynthesis, cytidine triphosphate: phosphocholine cytidylyltransferase, is post-translationally activated in response to the accumulation of free cholesterol in macrophages. This leads to an increase in phosphatidylcholine mass and the appearance of membrane whorls in these cells. We have advanced the hypothesis that this alteration in cellular phospholipid metabolism is an adaptive response to prevent the cellular free cholesterol: phospholipid ratio from reaching cytotoxic levels. Support for this hypothesis was obtained by demonstrating a direct relationship between the free cholesterol: phospholipid ratio and cellular necrosis in cultured macrophages, especially under conditions in which the phosphatidylcholine response was experimentally blunted. We propose that the eventual inability of this phospholipid response to keep up with free cholesterol accumulation in lesional macrophages in vivo may be an important cause of macrophage necrosis and, thus, plaque progression and clinical events in advanced atherosclerotic lesions. PMID- 9335948 TI - Cell-surface heparan sulfate proteoglycans: dynamic molecules mediating ligand catabolism. AB - Though sometimes regarded as merely passive, space-filling components, proteoglycans are in fact metabolically active molecules with carbohydrate and protein domains that are highly conserved throughout evolution, indicating specific, crucial functions. Here we review recent evidence that heparan sulfate proteoglycans, particularly syndecans and perlecan, are able to mediate directly the internalization of lipoproteins and other ligands, without requiring the participation of LDL receptor family members. Thus, heparan sulfate proteoglycans can function as receptors. In the case of syndecan heparan sulfate proteoglycans, efficient internalization is triggered by clustering of the transmembrane and cytoplasmic domains and then proceeds through a noncoated pit pathway, possibly caveolae. The physiologic and pathophysiologic importance of these direct heparan sulfate proteoglycan-mediated catabolic pathways in the liver and in the arterial wall in vivo remains to be settled. PMID- 9335950 TI - Mechanisms of oxidative damage of low density lipoprotein in human atherosclerosis. AB - Oxidatively damaged LDL may play a critical role in the pathogenesis of atherosclerotic vascular disease. Several pathways promote LDL oxidation in vitro but the physiologically relevant mechanisms have proven difficult to identify. Detection of stable compounds that result from specific reaction pathways has provided the first insights into the mechanism of oxidative damage in the human artery wall. Mass spectrometric analysis of protein oxidation products isolated from atherosclerotic tissue implicate tyrosyl radical, reactive nitrogen intermediates and hypochlorous acid in LDL oxidation and lesion formation in vivo. Hypochlorous acid is only generated by the phagocytic enzyme myeloperoxidase, which can also generate tyrosyl radical and reactive nitrogen intermediates. Chiral phase high-pressure liquid chromatography analysis of lipid oxidation products suggests that cellular lipoxygenases may also play a role at certain stages. In contrast, LDL isolated from atherosclerotic tissue is not enriched in protein oxidation products characteristic of free metal ions, which are the most widely studied in vitro model of LDL oxidation. These observations provide the first direct chemical evidence for reaction pathways that promote LDL oxidation in human atherosclerosis. PMID- 9335951 TI - The other side of scavenger receptors: pattern recognition for host defense. AB - Scavenger receptors bind modified lipoproteins and may play an important role both in normal and in pathological lipid metabolism. A number of different classes of scavenger receptors have been identified and several of these are multiligand receptors. Studies, both in vitro and in vivo, have indicated that at least some of these scavenger receptors may serve as pattern recognition receptors because they are able to bind a wide variety of pathogens. As a consequence, they may play key roles in innate immunity and host defense. PMID- 9335952 TI - Effects of estrogens on macrophage foam cells: a potential target for the protective effects of estrogens on atherosclerosis. AB - Estrogens have been shown to reduce markedly the development of atherosclerosis in humans and in several animal models. More than 50% of this effect appears to be caused by a direct action on the arterial wall. Although estrogens can affect a variety of functions of endothelial cells and smooth muscle cells, data published over the past few years suggest that macrophage foam cells may also be an important site of estrogen action. This review summarizes the recent data on the effects of estrogens on macrophage foam cell function. The data suggest that estrogens act in several ways to reduce the accumulation of cholesteryl esters in macrophages. PMID- 9335954 TI - Cell-mediated immunity in atherosclerosis. AB - Immunocompetent cells infiltrate atherosclerotic plaques of all stages. Plaque infiltrating T-cells recognize oxidized LDL and heat shock proteins and this elicits antibody responses that have been proposed as markers of disease activity. Cytokines secreted by activated T-cells may control macrophage activation, scavenger receptor expression and metalloproteinase secretion. Furthermore, cytokines secreted by T-cells and macrophages modulate smooth muscle proliferation, nitric oxide production and apoptosis, and induce endothelial activation. However, both positive and negative signals, as well as feedback loops, may be induced because of the complexity of the immune system. The possibility that some of these signals may be protective against atherosclerosis is currently under investigation in several laboratories. Recent studies in experimental animals show that immunomodulation affects the development of plaques and that immunization with oxidized LDL can inhibit lesion formation. This review provides a brief overview of cellular immunology and an analysis of its potential role in atherogenesis. PMID- 9335953 TI - Scavenging, signalling and adhesion coupling in macrophages: implications for atherogenesis. AB - Integrins are in physical association and functional cooperation with other membrane proteins that include receptors with scavenger functions, glycosyl phosphatidylinositol-linked receptors, the family of integrin associated multimembrane spanning signalling proteins and possibly other less characterized proteins with a coupling or signalling function. Monocyte adhesion, migration and differentiation to phagocytically active scavenger cells are directly coupled processes, involving integrins as common transducers for a panel of integrin linked specific receptors, which assemble a master cluster to coordinate adhesion, migration, scavenging and associated metabolic pathways of the lysosomal and secretory route, and also processes involved in host response. As macrophages represent highly heterogeneous cells that have major phenotypical and functional differences associated with specific patterns of integrin expression, the functional cooperation of integrins with scavenger receptors has to be related to specialized subsets of monocytes and macrophages as well as to ligand specific effects which mediate receptor coupling. PMID- 9335955 TI - Arterial calcification in face of osteoporosis in ageing: can we blame oxidized lipids? AB - Although vascular calcification is present in the vast majority of those aged over 70 years and causes serious cardiovascular dysfunction, strategies for its prevention and effective management are currently not available because the mechanism of its pathogenesis is unknown. Recently, similarities between mineralization in the vessel wall and in bone have been recognized. In this review, current understanding of the mechanisms involved in atherosclerotic calcification is summarized. In addition, several observations that may explain the paradox of vascular calcification in the face of osteoporosis are noted, emphasizing the possible role of lipid oxidation products. PMID- 9335956 TI - Novel members of the low density lipoprotein receptor superfamily and their potential roles in lipid metabolism. AB - The list of LDL receptor superfamily relatives is still growing. The two most recently discovered family branches are (1) a set of receptors characterized by expression in brain and a very close relationship to the LDL and VLDL receptors, and (2) highly cross-species (man, mouse, rabbit, chicken) conserved complex mosaic receptors which contain structural domains so far not found in the superfamily. At present, we know very little about the physiological function(s) of these molecules. They can be safely assumed, based on the presence of seven, eight or 11 clustered LDL receptor ligand binding repeats, to recognize mammalian apolipoprotein E but, based on the absence of apolipoprotein E in birds, they may exhibit interactions quite different from and/or in addition to typical lipoprotein receptors in vivo. PMID- 9335958 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 9335957 TI - Thrombosis and atherosclerosis. AB - The initial step in atherosclerosis is the rapid targeting of monocytes to the sites of inflammation and endothelial injury. Serum levels of intercellular adhesion molecule-1 were found to be increased in ischaemic heart disease patients and polymorphisms in the E-selectin gene were associated with accelerated atherosclerosis in young (age < 40 years) patients, further suggesting a role of inflammation in atherosclerosis. Cholesterol loading in macrophages was found to induce interleukin-8 expression, suggesting an association between foam cell formation and beta 2-integrin-dependent adhesion of leukocytes. Enhanced endothelium-platelet interaction induced by hypercholesterolaemia is mediated by von Willebrand factor, whereas platelet adhesion to subendothelial matrix is mediated by fibulin-fibrinogen complexes. Activated platelets mediate the homing of leukocytes by interaction with the subendothelial matrix under shear stresses that do not allow neutrophil adhesion. They may also contribute to the oxidative modification of LDL, provide a source of lipids for foam cell generation and contribute to smooth muscle cell proliferation. Oxidized LDL induces tissue factor in macrophages that also provide sites for fibrin polymerization and decreases the anticoagulant activity of endothelium by interfering with thrombomodulin expression and inactivating tissue factor pathway inhibitor. Intravascular fibrinolysis induced by tissue type plasminogen activator or urokinase may contribute to the initiation of atherosclerosis by inducing P-selectin and platelet activating factor as well as to plaque rupture, either directly or indirectly, by activating metalloproteinases. Plasminogen activator inhibitor-1 inhibits smooth muscle cell migration and, in the presence of vitronectin, promotes the clearance of thrombin by LDL receptor-related protein at sites of endothelial injury. PMID- 9335959 TI - Nutrition. Under-reporting in epidemiological and intervention studies: is identifying the under reporters enough? PMID- 9335960 TI - Genetics and molecular biology. PMID- 9335961 TI - Lipid metabolism. PMID- 9335962 TI - Hyperlipidaemia and cardiovascular disease. PMID- 9335963 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 9335964 TI - Therapy and clinical trials. PMID- 9335965 TI - Spatio-temporal patterns revealed in denoised fMRI data. PMID- 9335966 TI - Magnetic resonance imaging studies of functional brain activation: analysis and interpretation. AB - We have demonstrated that a time series of echoplanar images can contain low frequency noise components which confound analysis of functional MRI data. In simulated tasks of long duration, the false positive rate from t-test analyses greatly exceeded the statistical probability level. As task durations were shortened, the false positive rate declined. We also demonstrated that voxels representing extensive regions of the brain covary significantly over time. This covariation challenges the independence assumption of t-test and other analytical procedures and likely contributes to the false positive rate. The frequency spectra of many voxels showed relatively little power at higher frequencies with the important exception of some blood vessels (Fig. 12). Experimental designs in which stimulus or task conditions were alternated at these higher frequencies (e.g. 0.083 Hz corresponding to a 6 sec task duration and a 12 sec period for a complete two task cycle) did not show an inflated false positive rate when analyzed by t-test. We used the alternating tasks design with task durations of 8.73 sec, 6.4 sec, and 6.0 sec coupled with a frequency domain analysis strategy in a series of somatosensory, motor, perceptual, and working memory experiments. This combination of design and analysis was successful in identifying reliable activations across groups of subjects with a minimum of apparently spurious activations. By introducing a 180 degrees phase shift by reversing task order, we have been able to eliminate the contribution of most high frequency noise sources (such as large blood vessels). By segregating low frequency noise from the frequency of stimulus alternation, we routinely generate stable results in the presence of low frequency noise and drift. Despite the usefulness of the rapid task alternation and frequency domain techniques demonstrated here, there are potential problems and limitations in their application: 1. The short duration of our tasks results in an approximately sinusoidal activation waveform. With longer duration tasks, the activation time course would appear more square with a more complex frequency spectrum than the single peak demonstrated above. In such circumstances we have used convolution analysis with an expected waveform (McCarthy et al. 1996), similar to the approach of Bandettini et al. (1993). 2. If the activation in one task condition is significantly delayed and extends well into the period of the second task, it will be difficult to determine which task produced the activation. This problem is not specific to frequency analysis, and would occur as well for t-tests. One solution we have used is running a single active task against a relatively neutral control such as fixation to determine the usual activation dynamics of the active task. 3. Common activations by two alternating tasks are de-emphasized. This problem is also not specific to frequency analysis, and in most circumstances is an advantage rather than a disadvantage. However, if uncertain as to whether a task is capable of producing any activation, we have again used the strategy of running the task against a relatively neutral control. 4. Some tasks do not lend themselves to the short durations used here. 5. The frequency domain procedures used are conservative and may underestimate the true anatomical extent of the activation. In practice we compute t-tests in addition to the frequency domain techniques to guard against this possibility. Many of the advantages of the procedures described here are due to the alternation of short duration tasks rather than the application of frequency domain techniques per se. However, the success of these techniques in isolating periodic task-related signal changes suggest that a more complex design with concurrent stimulation presented at different frequencies might be feasible. Such designs may have advantages in that categories of stimuli would not be presented in isolation but against a changing ba PMID- 9335967 TI - What can we learn from MEG studies of the somatosensory system of the swine? PMID- 9335968 TI - MEG in the study of human cortical functions. PMID- 9335970 TI - High-frequency magnetic signals in the human somatosensory cortex. PMID- 9335969 TI - Comparison of functional localization in human visual cortices using MEG and fMRI: a preliminary report. PMID- 9335971 TI - MEG imaging of neuronal population dynamics in the human thalamus. PMID- 9335972 TI - Correspondence of anatomy and function in the human digit sensory cortex revealed by an MRI-linked whole-head MEG system. PMID- 9335973 TI - Localization of somatosensory evoked P22 component by magnetoencephalography. PMID- 9335974 TI - Neuromagnetic analysis of sensory-motor hand areas in man. PMID- 9335975 TI - Pain-related brain responses following CO2 laser stimulation: magnetoencephalographic studies. PMID- 9335976 TI - MEG measurements of 40 Hz auditory evoked response in human brain. PMID- 9335977 TI - Right hemispheric dominance in the auditory evoked magnetic fields for pure-tone stimuli. PMID- 9335978 TI - Trial measurements of gustatory-evoked magnetic fields. PMID- 9335979 TI - Measurement of olfactory event-related magnetic fields evoked by odorant pulses synchronized with respiration. PMID- 9335980 TI - Voluntary attention to the motion of visually perceived objects can specifically activate either V1 or MT. PMID- 9335981 TI - Neural substrates of working memories are revealed magnetically by the local suppression of alpha rhythm. PMID- 9335982 TI - Studies on integrative functions of the human frontal association cortex by use of MEG. PMID- 9335983 TI - Neural representation of concurrent sounds: a magnetoencephalographic study. PMID- 9335984 TI - Analysis of magnetic signals related to reading Japanese characters (hiragana). PMID- 9335986 TI - Event-related magnetic fields during processing of readable and unreadable character strings. PMID- 9335985 TI - Cerebral activities of the human brain in a delayed matching task of visual characters. PMID- 9335987 TI - Missing auditory stimuli activate the primary and periauditory cortices: combining MEG and fMRI studies. PMID- 9335988 TI - Brain activation during learning of sequential procedures. PMID- 9335989 TI - High resolution cortical activation mapping of voluntary eye movements using functional magnetic resonance imaging. PMID- 9335990 TI - Activation of human primary and secondary motor areas during imagination of and actual finger movement using echo planar imaging. PMID- 9335991 TI - Comparison of supplementary motor area activation between simple and complex motor task. PMID- 9335992 TI - Estimation of movement-related brain activities with visual stimuli. PMID- 9335993 TI - Cerebral activities related to accommodation: a neuromagnetic study. PMID- 9335994 TI - Human voluntary movement physiology as studied by DC-EEG, MEG, SPECT and FMRI. PMID- 9335995 TI - Functional MRI and magnetic resonance spectroscopy with clinical MRI scanners. PMID- 9335996 TI - Cortical mapping using an MRI-linked whole head MEG system and presurgical decision making. PMID- 9335997 TI - Neurological application of MEG. PMID- 9335998 TI - Current source localization of EEG paroxysms in a patient with congenital mirror movement. PMID- 9336000 TI - Epileptic events observed by multichannel MEG. PMID- 9335999 TI - Neuropsychiatric applications of MEG. PMID- 9336001 TI - Generators of rolandic discharges identified by magnetoencephalography. PMID- 9336002 TI - Development of a multi-channel SQUID system for magnetoencephalogram applications. PMID- 9336003 TI - A 256-channel magnetometer for brain research. PMID- 9336004 TI - Reconstruction of sparse dipole source by a modified SMN method. PMID- 9336005 TI - Inverse problem of traveling electrical sources in the human brain associated with auditory evoked magnetic fields. PMID- 9336006 TI - Combined 3D neuromagnetic source imaging and MRI-scans. PMID- 9336007 TI - Analysis and visualization of MEG data with the application visualization system (AVS). PMID- 9336008 TI - Commercially available luminometers and imaging devices for low-light level measurements and kits and reagents utilizing bioluminescence or chemiluminescence: survey update 5. AB - This survey was compiled in July 1997 and includes products not covered in the luminometer survey (Jan 1992: Stanley PE, J Biolumin Chemilumin 1992; 7:77-108 and 7:157-69), kits and reagent survey (Nov 1992; Stanley PE, J Bilumin Chemilumin 1993; 8:51-63), update 1 (June 1993: luminometers, kits and reagents, Stanley PE, J Biolumin Chemilumin 1993; 8:237-240) and update 2 (Dec 1993: luminometers, kits and reagents, Stanley PE, J Biolumin Chemilumin 1994; 9:51-3) and update 3 (Feb 1994: luminometers, kits and reagents, Stanley PE, J Biolumin Chemilumin 1996; 11:175-91). Technical details are provided together with company address and contact information including email and website where known. Items include: Luminometers, radiometers, low-light imaging, CCD cameras, immunoassays, ATP rapid microbiology, hygiene monitoring, molecular probes, labels, nucleic acid hybridization, reporter genes. PMID- 9336009 TI - Interaction of trout hemoglobin with H2O2: a chemiluminescence study. AB - Erythrocytes from trout Salmo irideus are characterized by four different hemoglobin components (HbI, HbII, HbIII and HbIV), HbI and HbIV being predominant. In this study we describe the interaction between trout hemoglobin (HbI and HbIV) and H2O2 using a chemiluminescence assay. Our data show that the reaction of hemoglobins with H2O2 produces a time-limited and significant increase of chemiluminescence signal. The half-life of the decay of this chemiluminescence signal was characteristic for each type of hemoglobin used. These results indicate the formation of excited molecules related to the interaction between trout hemoglobin and H2O2. PMID- 9336010 TI - The widespread occurrence and tissue distribution of the imidazolopyrazine luciferins. AB - Bioluminescence has been reported to occur in 17 phyla and at least 700 genera. However, the luciferin chemistry of the majority of luminous organisms has yet to be determined. The most common chemistry which is known to occur in deep sea bioluminescence is imidazolopyrazine bioluminescence. The main aim of this study was to examine the phyletic and tissue distribution of imidazolopyrazine luciferins. This will facilitate analysis of imidazolopyrazine bioluminescence at the cellular and molecular levels and, in particular, how and when its chemistry is controlled and expressed in vivo. Assays for both known imidazolopyrazines were established and a range of fresh organisms and tissue were analysed, i.e. fish, cephalopods, copepods, ostracods, amphipods and euphausiids. The main findings were that the number of genera in which coelenterazine has been detected has been increased from 52 to about 90. Also, for the first time, the other known imidazolopyrazine luciferin, Vargula-type luciferin, was quantified in the ostracod Cypridina dentata, but was not detected in any of its potential predators. Neither imidazolopyrazine luciferin was found in several luminous stomiiform fish assayed. Coelenterazine was measured in the livers and photophores of a number of cephalopods and it is apparent that coelenterazine is responsible for both modes of luminescence. PMID- 9336011 TI - Bioluminescence and chemiluminescence literature. Green fluorescent protein. PMID- 9336012 TI - Structure and mechanism of carbonic anhydrase. AB - Carbonic anhydrase (CA; carbonate hydro-lyase, EC 4.2.1.1) is a zinc-containing enzyme that catalyzes the reversible hydration of carbon dioxide: CO2+ H2O<- >HCO3(-)+H+. The enzyme is the target for drugs, such as acetazolamide, methazolamide, and dichlorphenamide, for the treatment of glaucoma. There are three evolutionarily unrelated CA families, designated alpha, beta, and gamma. All known CAs from the animal kingdom are of the alpha type. There are seven mammalian CA isozymes with different tissue distributions and intracellular locations, CA I-VII. Crystal structures of human CA I and II, bovine CA III, and murine CA V have been determined. All of them have the same tertiary fold, with a central 10-stranded beta-sheet as the dominating secondary structure element. The zinc ion is located in a cone-shaped cavity and coordinated to three histidyl residues and a solvent molecule. Inhibitors bind at or near the metal center guided by a hydrogen-bonded system comprising Glu-106 and Thr-199. The catalytic mechanism of CA II has been studied in particular detail. It involves an attack of zinc-bound OH- on a CO2 molecule loosely bound in a hydrophobic pocket. The resulting zinc-coordinated HCO3- ion is displaced from the metal ion by H2O. The rate-limiting step is an intramolecular proton transfer from the zinc-bound water molecule to His-64, which serves as a proton shuttle between the metal center and buffer molecules in the reaction medium. PMID- 9336014 TI - Modulation of immune cell function by the autonomic nervous system. AB - This review discusses some of the major findings implicating the autonomic nervous system in the regulation of immune function. The sympathetic nervous system, the primary focus of this line of research, directly innervates the major lymphoid organs, and physiological release of sympathetic neurohormones at these sites has been documented. Leukocytes have been shown to express receptors for catecholamines, as well as neuropeptide Y, and studies in vitro and in vivo have indicated that occupation of these receptors by the appropriate ligands produces functional changes in immunological cells. Finally, altered sympathetic regulation may underlie some of the immunological abnormalities observed in chronic stress, clinical depression, and ageing. PMID- 9336013 TI - Nicotine for the treatment of Tourette's syndrome. AB - Recent evidence has demonstrated that nicotine may obtund the symptoms of Tourette's syndrome (TS). TS is a neuropsychiatric disorder characterized by motor and vocal tics, obsessions and compulsions, and frequently with impulsivity, distractibility, and visual-motor deficits. While neuroleptics, such as haloperidol, are most effective for treatment of the motor and vocal tics of TS, these medications have many side effects. In this article, we review the evidence, consistent with findings in animals, that administration of nicotine (either 2 mg nicotine gum or 7 mg transdermal nicotine patch) potentiates the therapeutic properties of neuroleptics in treating TS patients and that a single patch may be effective for a variable number of days. These findings suggest that transdermal nicotine could serve as an effective adjunct to neuroleptic therapy for TS. PMID- 9336015 TI - Therapeutic drug monitoring opportunities in cancer therapy. AB - Cancer is a common cause of death, and improvements in treatment are desperately needed. The high degree of variation in systemic exposure for a given dose and the relationships between blood concentrations and either toxic or antitumor effects would suggest that therapeutic drug monitoring is a potential mechanism for improving the treatment of individual patients. In this review, the case for therapeutic drug monitoring is made in a select number of commonly used cancer drugs and areas that require more concerted effort are highlighted. PMID- 9336016 TI - Low-density lipoprotein and oxidised low-density lipoprotein: their role in the development of atherosclerosis. AB - Oxidation of low-density lipoprotein (LDL) may be implicated in the development of atherosclerotic disease. Oxidised LDL is taken up more readily by monocyte derived macrophages than LDL. Antibodies to oxidised LDL are found in atherosclerotic lesions, Increased risk of ischaemic heart disease is associated with a preponderance of small dense LDL particles, which are more susceptible to oxidation. Proatherogenic alterations in cell biochemistry and signalling pathways occur in the presence of LDL and more markedly oxidised LDL. In vitro antioxidants inhibit changes in cell biochemistry, while in vivo, they have been shown to attenuate or reverse development of atherosclerosis. PMID- 9336017 TI - The N-formyl peptide receptor: a model for the study of chemoattractant receptor structure and function. AB - N-formyl peptides, such as fMet-Leu-Phe, are one of the most potent chemoattractants for phagocytic leukocytes. The interaction of N-formyl peptides with their specific cell surface receptors has been studied extensively and used as a model system for the characterization of G-protein-coupled signal transduction in phagocytes. The cloning of the N-formyl peptide receptor cDNA from several species and the identification of homologous genes have allowed detailed studies of structural and functional aspects of the receptor. Recent findings that the receptor is expressed in nonhematopoietic cells and that nonformylated peptides can activate the receptor suggest potentially novel functions and the existence of additional ligands for this receptor. PMID- 9336018 TI - Inhibition of Ras prenylation: a novel approach to cancer chemotherapy. AB - The demonstration that Ras requires prenylation for its cancer-causing activity led several groups of investigators to an intense search for farnesyltransferase and geranylgeranyltransferase inhibitors as potential anticancer drugs. Rational design of small organic molecules that mimic the carboxyl terminal tetrapeptide prenylation site on Ras resulted in pharmacological agents capable of inhibiting Ras processing and selectively antagonizing oncogenic signaling, and suppressing human tumor growth in mouse models without side effects. These agents presently are undergoing advanced preclinical studies. This review describes the efforts of several groups to design, synthesize and evaluate the biological activities of several classes of prenyltransferase inhibitors. Several important issues, such as mechanism of action of prenyltransferase inhibitors and potential mechanisms of resistance to inhibition of K-Ras farnesylation, are also discussed. PMID- 9336019 TI - Purinoceptors in neuromuscular transmission. AB - At the neuromuscular junction, P2-purinoceptors mediate the actions of the co transmitter ATP and P1-purinoceptors, those of its degradation product adenosine. The classification of the subtypes of P1- and P2-purinoceptors and their signal transduction routes is presented. Purinoceptor-mediated effects on the prejunctional release of acetylcholine and the postjunctional desensitization and expression of nicotinic receptors are discussed in depth. An additional section on the reversal action of the P2-purinoceptor antagonist suramin on neuromuscular block underscores the importance of testing purinoceptor-targeted drugs once they will be marketed, to avoid adverse effects in patients. PMID- 9336021 TI - Losartan, an orally active angiotensin (AT1) receptor antagonist: a review of its efficacy and safety in essential hypertension. AB - Losartan potassium, an angiotensin II receptor antagonist, is the first of a new class of agents to be introduced for the treatment of hypertension. In this review, we describe the clinical pharmacology of losartan, including its pharmacokinetics in healthy, male volunteers and special patient groups, such as the elderly, patients with liver disease and patients with renal impairment. We also review its pharmacodynamics, including safety and tolerability; specificity of action; and the effect of salt depletion. We then review the studies examining clinical efficacy and safety in hypertension. PMID- 9336022 TI - DNA vaccines. AB - In just a few years, injection of plasmid DNA to elicit immune responses in vivo has developed from an interesting observation to a viable vaccine strategy. DNA vaccines have been shown to elicit both cellular and humoral immune responses and to be effective in a variety of preclinical bacterial, viral, and parasitic animal models. This review will discuss the current knowledge of vector design, methods of plasmid delivery, immune responses elicited by various DNA vaccines, safety issues, and production and release of plasmid as a vaccine product. The potential of this new vaccine strategy and its future prospects is summarized. PMID- 9336020 TI - Pharmacology of cannabinoid CB1 and CB2 receptors. AB - There are at least two types of cannabinoid receptors, CB1 and CB2, both coupled to G-proteins. CB1 receptors are present in the central nervous system and CB1 and CB2 receptors in certain peripheral tissues. The existence of endogenous cannabinoid receptor agonists has also been demonstrated. These discoveries have led to the development of selective cannabinoid CB1 and CB2 receptor ligands. This review focuses on the classification, binding properties, effector systems and distribution of cannabinoid receptors. It also describes the various cannabinoid receptor agonists and antagonists now available and considers the main in vivo and in vitro bioassay methods that are generally used. PMID- 9336023 TI - Cytotoxic therapy and pregnancy. AB - The use of cytotoxic agents during pregnancy may be unavoidable in order to ensure maternal survival-despite the dangers to the developing fetus. We review 217 such cases published between 1983 and 1995, classifying them into 5 groups according to whether the cytotoxic drugs were used to treat leukaemias, malignant lymphomas, severe rheumatic diseases, gynaecological/breast neoplasms, or other grave conditions. Various factors, such as the drug type, dose, and timing to exposure to gestational age, are analysed with respect to the outcome of these pregnancies (teratogenicity, stillbirths, spontaneous abortions, prematurity, etc.). These results are then integrated in order to determine whether one can predict the individual risk of abnormality for the newborn when cytotoxic agents must be administered to pregnant women faced with a malignancy or other serious condition. PMID- 9336024 TI - Drug management of hypertensive disorders of pregnancy. AB - Drugs used in the acute and long-term management of hypertension in pregnancy and the preeclampsia-eclampsia syndrome have been reviewed and their therapeutic effects and maternal and fetal adverse effects have been considered. The review also focuses on recent developments in the areas of prevention and management of pre-eclampsia-eclampsia syndrome. Although a number of new drugs have emerged, as potentially useful in the management of hypertension in pregnancy and pre eclampsia-eclampsia syndrome, some remain at the cornerstone of therapy; for example, methyldopa for long-term treatment of chronic hypertension, hydralazine or nifedipine for rapid reduction of severely elevated blood pressure, and magnesium sulphate for eclampsia. Some of these agents, especially the calcium antagonists, show promise in that their use is associated with fewer side effects. Safety for the fetus, however, has not been adequately evaluated yet. Neither aspirin nor calcium supplements appear to improve the outcome in pregnancy. Currently, the dilemma whether to treat hypertension in pregnancy and pre-eclampsia-eclampsia syndrome with old, established, cost-effective drugs or the promising newer drugs provides an interesting academic challenge. PMID- 9336040 TI - [Hibernating myocardium--a reality?]. PMID- 9336041 TI - [Welcome to research activities in today's Brazilian medical schools]. PMID- 9336042 TI - [Cryopreservation of bone marrow and peripheral blood stem cells using a controlled rate freezing system: experience with 86 procedures]. AB - The cryopreservation of hematopoietic stem cells can be used for rescuing the hematopoiesis after high dose chemotherapy. PURPOSE: The ice crystal formation during the freezing procedure is the key point that can be harmful to the cells. The cryopreservation of hematopoietic stem cells in a controlled-rate freezer could decrease the cell damage. METHODS: Twenty-three patients with a median age of 26 years (range 03-57) had bone marrow and/or peripheral blood stem cells harvested from March 1993 through October 1994, ending up to 86 freezing procedures. The patient's diagnoses are as follows: Non-Hodgkin's Lymphoma (n = 5); Acute Myelogenous Leukemia (n = 8); Acute Lymphocytic Leukemia (n = 6); Hodgkin's disease (n = 3); Multiple Myeloma (n = 1). The cells were frozen away in a controlled-rate freezer chamber at the following rate: -1 degree C/min from room temperature to -45 degrees C and then, at -10 degrees C/min down to -80 degrees C. After freezing, the cells were kept into mechanical freezers until the marrow infusion. To mobilize PBSC (peripheral blood stem cells), G-CSF (granulocyte colony stimulating factor) was given. RESULTS: A median of 3.16 x 10(8) cells/kg (range 0.86-24.22) of PBSC and 2.03 x 10(8) cells/kg (0.19-12.21) of bone marrow cells were frozen. The median time to reach granulocytes greater than 500/microL and platelets greater than 20,000/microL was 12 days (range 8-40) and 31 days (range 8-80), respectively. All patients had marrow engraftment after infusion of hematopoietic stem cells. CONCLUSION: The cryopreservation procedure using a controlled-rate freezer can store hematopoietic stem cells and potentially, cause less damage to the cells. PMID- 9336044 TI - [Monomeric plasmatic calcitonin and hypercalcemia in lung cancer patients]. AB - BACKGROUND: Calcitonin (CT) is a peptidic hormone produced by the thyroid C cells and related to calcium metabolism. High plasmatic levels of this hormone are found in patients with medullary thyroid carcinoma, what makes it an excellent tumor marker for this disease. However, there are reports that showed an increase of plasmatic CT levels in patients with other tumors, mainly in lung cancer. PURPOSE: These data prompt us to investigate the validity of the CT level determinations as a potential tumor marker in different histologic lung cancer, and its correlation with hypercalcemia, a very common complication in these tumors. METHOD: Blood were sampled from 56 patients with malignant lung disease for the CT and ionized calcium determinations. Calcitonin was measured using a specific radioimmunoassay for the monomeric form of the molecule, in a previous silica extracted serum probe. RESULTS: We did not find elevated levels of monomeric CT in lung cancer. Only 3 patients had mild elevated levels, while in the others CT was normal or undetectable. Hypercalcemia was found in 21.4% of these patients, but only one with supranormal CT levels. CONCLUSION: Monomeric CT serum levels are normal in lung cancer, what makes the latter use an unreliable tumor marker. PMID- 9336045 TI - [Assessment of body composition in pregnant women at term]. AB - BACKGROUND: Pregnancy is associated with well-known physiologic changes of maternal fluid and energy compartments, along with organ hypertrophies and the appearance of fetal and placental tissues. As a consequence, body composition is modified, but this phenomenon has not been well documented. The advent of bioimpedance has contributed to the documentation of the desired information in a safe and practical way. AIMS: The aims of this study were: To register the principal anthropometric variables in a population of pregnant women at term; To determine body composition by bioimpedance analysis; To compare these findings with the former results, as well as with other assessment procedures reported in the literature. PATIENTS AND METHOD: The population consisted of 30 pregnant women at term admitted for delivery, without complications, fetal distress or multiple pregnancy, and submitted to the following measurements: Anthropometry- Weight, height, body mass index, triceps skinfold, arm muscle circumference; Bioimpedance analysis--Body fat, lean body mass, total water, intra and extra cellular water, third space, and exchangeable Na/K ratio. RESULTS: Body weight and body mass index were increased but within the expected values for these patients. Total body water was similar to results in non-pregnant women when expressed as percentage, in accordance with other studies, but with a trend toward increase in the extracellular compartment and presence of third space fluid. Body fat was elevated, but the proportions were not much different from previous anthropometric surveys. CONCLUSIONS: In the conditions of this investigation, in which a bioimpedance equation for general use was employed, the method indicated results that were consistent with the clinical course, anthropometric documentation, and the findings of other groups. It is concluded that bioimpedance analysis compares favorably with other assessment procedures in pregnancy, and further studies with this method should be undertaken. PMID- 9336043 TI - [Comparison of preschool children's performance using the Denver developmental test, before and after nutritional intervention]. AB - Psychomotor and development analysis must be emphasized when studying institutionalized children. Many previous investigations have been showing deleterious effects of day care centers over developmental performance in children. OBJECTIVE: This study is aimed at comparing the performance in the Development Screening Test (Denver) in children attending day care centers, before and after nutritional intervention with an energetic supplement enriched with iron. METHOD: 130 children from 2 to 6 years old, attending three municipal day care centers, were evaluated by means of the application of the Denver test; by trained psychologists, comparing the collected data according to sex and age group, before and after six months intervention with iron enriched protein energetic supplement. RESULTS: Most of the children had normal performance, both in first application (70.80%), and in the second one (80.80%). When comparing these results, 76.92% of the children had not altered their performance and 18.46% improved it substantially. As to sex, no significant differences were found and as to age group, there was significant improvement among children aged 4 to 6 years of age. CONCLUSIONS: Besides the nutritional aspects, factors such as learning readiness, family organization, and psychopedagogic orientation to the day care centers, must have fostered development, even if the low socioeconomic level of the studied population is considered. PMID- 9336046 TI - [Predictive value of the measurement of iodothyronines in the prognosis of patients with severe nonthyroidal illness]. AB - PURPOSE: In order to find prognostic parameters in patients with severe diseases, we analyzed sequentially the levels of thyroid hormones. METHODS: We measured iodothyronines (T3, T4 and rT3) in 42 patients before the admission and after the discharge in an intensive care unit. In addition, we also measured the iodothyronines in other 17 patients after the discharge. RESULTS: Comparing the group of good outcome with the patients who died, we observed in the former group initial normal T4 levels in 76% of the patients, which were maintained in 65% of them during hospitalization and in 70% of them at the time of delivery from the intensive care unit. Patients who died, however, presented initial low T4 levels in 56% of them, decreasing values in 95% of them during hospitalization and low levels in 81% of patients at the last dosage. The combined profile of T3 and T4 also differentiated good and bad outcome. CONCLUSION: We suggest that serial analysis of serum levels of thyroid hormones may help the evaluation of critical care patients. PMID- 9336047 TI - [Deep neck spaces and their significance in cervical infections]. AB - BACKGROUND: Although rare, deep neck space infections are associated with high morbidity and mortality rates. The surgical approach is necessary in the majority of the cases, and the surgeon must know the complex anatomy of the cervical fasciae and deep neck spaces. PURPOSE: The anatomy of the cervical fasciae and deep neck spaces in reviewed. As an illustration, a series of deep neck space infections is presented. MATERIAL AND METHOD: Four clinical cases are reported: 1) a case of Ludwig's angina with several complications (mediastinitis, pericarditis, pneumonia, pleural effusion and empyema, esophageal fistula and septic shock), 2) a case of cervical abscess that appeared without apparent cause, in a young diabetic patient, 3) a case of abscess of the submandibular triangle, and 4) a case of parapharyngeal abscess that came forth after a dental treatment. Data from history taking, physical examination, X-rays, echography, CT scan and treatment and the follow-up are presented. The image tests were valuable and, in two of the cases, they demonstrated that more than one deep neck space were affected. CONCLUSIONS: The literature reinforces the high mortality and morbidity rates, the diversified etiology (dental infection, intravenous drug abuse, infections of the upper aerodigestive tract and others), and the tracheostomy indication made in about half of the cases. It stresses also the need for combined therapy (antibiotics and surgery). Evaluation with CT scan and other radiologic methods is indispensable to determine the site and extent of the process and to plan properly the treatment. PMID- 9336048 TI - [Vertebral and femoral bone mineral density of 724 caucasian Brazilian women: influence of age and body weight]. AB - OBJECTIVE: To study the vertebral (L2-L4) and femoral (neck) bone mineral density (BMD) of normal white women. MATERIAL AND METHOD: We measured the BMD of 724 women (40-79 kg; 20-69 years-age) by dual-energy X-ray absorptiometry. Data were analysed as a function of age and body weight (BW). RESULTS: Thinner women (40-49 kg) attained maximal vertebral and femoral BMD (mBMD) at ages between 30-39 years, while heavier women (60-79 kg) already had the mBMD by the age of 20. At the femur, there was a significant mBMD-BW correlation (r = 0.97; p < 0.001; slope = 0.72%/kg). At the spine, only the 40-49 Kg women exhibited lower mBMD when compared to the others (p < 0.001). The decrease of the vertebral BMD was more intense (-8.3 vs. -5.7%/decade) and started earlier (fourth vs. fifth decade) in women weighting 40-59 kg, as compared to those weighting 60-79 kg. The decrease of the femoral BMD was initiated just after mBMD was achieved and, at the age of 69, heavier women showed a decrease that was 5.3% lower than those weighting 40-49 kg. The vertebral BMD of the Brazilian women was practically the same as reported for a North-American population. CONCLUSIONS: (i) Vertebral and femoral BMD of this Brazilian population varied with age similarly to other white female populations; (ii) provided that appropriate corrections are made for BW, the BMD of Brazilian women is comparable to the BMD of North-Americans; and (iii) the BW is important both in acquisition and decline of bone mass, as it influences the relation BMD-age. PMID- 9336049 TI - [Evaluation of in vitro activity of new fluoroquinolones, cephalosporins and carbapenems against 569 gram-negative bacteria]. AB - OBJECTIVE: Evaluation of the in vitro activity of new fluoroquinolones, cephalosporins and carbapenems against gram-negative bacteria. MATERIAL AND METHOD: A total of 569 clinical isolates were obtained from inpatients at Sao Paulo Hospital--UNIFESP/EPM in June and July of 1992. The species distribution was as follows: Enterobacter sp. (62), Escherichia coli (308), Klebsiella pneumoniae (27), Klebsiella sp. (9), Proteus mirabilis (23), Pseudomonas aeruginosa (88), Pseudomonas sp. (4), Serratia sp. (30) and other gram-negatives (7). Susceptibility tests were performed by broth microdilution. The antimicrobials agents tested were: ciprofloxacin, ofloxacin, levofloxacin, grepafloxacin, DU 6859-alpha, ceftazidime, cefepime, FK 037, imipenem, meropenem and biapenem. RESULTS: DU 6859-alpha showed the highest anti-microbial activity among the fluoroquinolones. It was two- to four-fold more active than ciprofloxacin against some species. The potency and antimicrobial spectrum were similar between the fourth-generation cephalosporins against Enterobacteriaceae, except for Enterobacter sp. strains which were more susceptible to cefepime than they were to cefetazidime or FK 037. When testing Pseudomonas aeruginosa, ceftazidime was slightly more active than the other cephalosporins. Against Enterobacteriaceae and Pseudomonas aeruginosa strains, meropenem was more active than imipenem or biapenem. In addition, the percentage of strains, susceptible to meropenem was higher than the percentage susceptible to the other cerbapenems against these species. CONCLUSION: The new antimicrobial agents demonstrated in vitro activity higher than that of agents commercially available. However, more studies are necessary to further evaluate the in vivo activity and the clinical benefit of these compounds. PMID- 9336050 TI - [Effects on the pulmonary function after single dose of exogenous pulmonary surfactant in children with acute respiratory distress syndrome]. AB - The Acute Respiratory Distress Syndrome (ARDS) is a pulmonary lesion of multifactorial cause in which the surfactant system is altered owing to inactivation and impairment of composition and metabolism. The use of exogenous pulmonary surfactant is a therapeutic option with the objective to maintain alveolar stability thus improving the pulmonary compliance (increasing the residual functional capacity), oxygenation and ventilatory mechanics. A study carried out on two pediatric patients with ARDS submitted to mechanic pulmonary ventilation, applying a single dose of exogenous pulmonary surfactant is described. The patients were evaluated using arterial and venous gasometry before and after the use of surfactant, observing increment in oxygenation, reduction of shunt fraction, improvement in ventilation immediately after exogenous pulmonary surfactant instillation and return to the previous situation after 240 minutes in case 1 and 120 minutes in case 2. More prospective clinical and randomized studies are needed to effectively evaluate this therapeutic modality. PMID- 9336052 TI - [The hibernating myocardium an its recognition]. PMID- 9336051 TI - [Depressor anguli oris myocutaneous island flap for total upper lip and columella reconstruction]. AB - Case report of a man that had been previously submitted to many resections of facial skin carcinomas. A serious deformity on the upper lip and columella remains. A total upper lip and columella reconstruction through the depressor anguli oris myocutaneous island flap is described. PMID- 9336053 TI - [Diabetic gastroparesis]. PMID- 9336054 TI - [How to create a smoke-free hospital]. PMID- 9336055 TI - [Delayed intra-uterine growth: current aspects]. PMID- 9336056 TI - [Effects of parathion on acetylcholinesterase activity in rat kidney]. AB - In this work the effects of parathion (a competitive acetylcholinesterase inhibitory agent) on the enzyme activity was studied by cytochemical methods in the kidney of rats; being the doses used not inhibitory of red blood cells acetylcholinesterase (subtoxic dose). Two groups of rats were used: control (CR) and parathion-treated (TR) groups, both submitted to a water deprivation period of 24 h. for induction of primary thirst. The treated-rats group received parathion i.p. at doses of 600 micrograms/100g body weight. For demonstration of acetylcholinesterase activity, renal tissue incubation was performed by the Karnovsky and Roots method with the Tsuji Larabi variation. The results of the five assays of incubation enable us to conclude that there exists a noticeable activity of acetylcholinesterase in the renal cortex of control rats, which is blocked by subtoxic doses of organophosphorus pesticide. We suggest that the natriuretic effect of parathion can be explained by this mechanism. PMID- 9336057 TI - [Design of a thermic detection system applied to chick embryonated eggs irradiated with infrared rays]. AB - Infrared radiations are widely used in medical therapeutics. It has been argued that the doses and the periods of time employed in experimental animals are higher than those used in clinic. Thus, we considered of interest to analyse aspects of dosimetry and thermic effects of infrared rays with current methods in medical practice, using the in ovo chick embryo as a model of easy control. To this end we designed a system to measure temperatures and their acquisitions and software for its handling. The system consists of: a) thermic points: thermocuples or termistores adaptable to the experimental requirements and calibrated with a greater precision within a range of ten degrees around the incubation temperature; b) acquirer circuit of thermic data (hardware): it generates a time base that varies with the thermic sensor. Software: the PC XT or AT detects changes in the time base by means of a programs' in a Turbo Pascal; c) storage and analysis of data allows, through a menu (expansibles) the scale selection, time of program data to be acquired, storage and recovery of the diskette information and graphic impression; d) chick's embryonated eggs. This system allows to measure temperature distribution in small physical spaces with little disturbance of the system to be measured in irradiated bodies, to analyse variations of the temperatures in time and to secure a greater confidence and automatism to obtain the required data. PMID- 9336058 TI - [Immune complex detection in nasal mucosa of patients with chronic and permanent nasal obstruction]. AB - Thirty patients with rhinologic problems and in which the main symptom was "Permanent Nasal Blockade" were studied. We found that they presented a "Permanent and Total Chronic Nasal Blockade"-an entity defined by us as a permanent chronic obstruction of the nasal cavities that would be the clinical and anatomopathologic expression and the immunological terminal phase of "Chronic Rhinitis", leading to Chronic Respiratory Nasal Insufficiency. The main symptoms of these "buccal breathers" were: throat dryness, night snores, nasal resonance of the voice, mechanic and secondary disturbances derived from mouth breathing, such as pharingitis, laryngitis, bronchitis, bronchial asthma and a tendency towards emphysema; dystrophia of the facial bones, disminution of the intellectual capacity, irritability, lack of concentration; disturbances of the circulatory function (alterations of the cardiac rhythm and of the blood pressure). The patients were adolescents and young adults in which allergic and infectious causes dominated the etiology, showed by family allergic and personal atopic antecedents, positive intradermic tests for aeroallergenes and bacteria and corroborated by increase of immunoglobulin E and of blood eosinophiles. The methods of diagnosis used to verify the "Permanent and Total Chronic Nasal Blockade" were: X-rays, lineal tomography; rhinomanometry and rhinofibrolaringoscopy. The anatomopathologic results obtained by nasal biopsy showed a lymphomonocitary infiltrate around arterioles and venules: signs of vasculitis. All this led us to search for immunocomplexes. The presence of positive immunocomplexes surrounding the vessels contributed to a more precise focusing of the physiopathology, diagnosis and treatment of "Permanent and Total Chronic Nasal Blockade". PMID- 9336059 TI - [Primary malignant esophageal melanoma: report of a case]. AB - Primary malignant melanoma of the esophagus is rare. We identified one patient over a period of 15 years. This patient was a 80 years-old caucasian man. No association was found with tobacco or ethanol use, nor was there a personal or family history of malignant melanoma. Symptoms were related to obstruction. This tumor was polypoid in its upper part and ulcero-infiltrant in its lower part. Histologically the melanoma had epithelioid spindle cells. The neoplasm was immunoreactive for S-100 protein and non reactive with anti-cytokeratins. This patient was treated by Garlock type, esophagectomy with excision of 13 cm of esophagus and 2 cm of stomach. The survival was of only 3 days, because he developed acute respiratory and cardiac disease syndrome and died. PMID- 9336060 TI - [A case of ovarian lipoid cell tumor: histopathology]. PMID- 9336061 TI - [Stunting risk factors in 5-year-old children]. AB - A retrospective study of ninety two boys and one hundred and four girls at the age of five years was made with the purpose of identifying Stunting Risk Factors. "Case" was defined as a five years-old child/with height/age < 95% of the median NCHS norm and "control" as a child with height/age > 95% of the median of the same norm. Seventy six biological, psicoaffective, nutritional and socioeconomic cultural variables were analysed on the children and their families. Data were obtained from interviews and clinical histories' revision. Odds Ratio, at a P confidence interval of 95%, Mantel-Haenszel x2, Wolf Test and Anova were calculated. Significative risk factors were: masculine sex, mother height < 1.50 cm, intergenesic interval less than 33 months referred to the immediate elder born, negligent mother, numerous family, crowded family, unadequate excrete disposition and having one undernourished brother or sister. Variance analysis also showed significant differences for: birth height, negligent mistreaing, physical mistreaing, repeated parasitic episodes, and birth order superior to third son. PMID- 9336062 TI - [Molecular biology of chronic myeloid leukemia]. PMID- 9336063 TI - [Metalloproteinase activity in arteries and veins. Regulation with doxycycline]. AB - BACKGROUND: Alterations in the synthesis and degradation of extracellular matrix occur during atherogenesis. Metalloproteinases, whose activity may be inhibited with doxycicline in other tissues, play an important role in this process. AIMS: 1. To characterize metalloproteinase activities in internal mammary artery and saphenous vein, and 2. To assess the effect of doxycicline in the activity of metalloproteinases of these vessels and of cultured smooth muscle cells. METHODS: Segments of internal mammary arteries and saphenous veins and cultured smooth muscle cells were incubated with and without doxycicline. Metalloproteinases activity was assessed by zymography and Western Blot. RESULTS: Activity of metalloproteinase-9 in saphenous veins was 217% less than in internal mammary arteries. In these vessels doxycicline decreased metalloproteinase-9 activity by 207% and metalloproteinase-2 by 290%. Western Blot analysis showed that docycicline also inhibited metalloproteinase-1 expression. In cultured smooth muscle cells, the median inhibitory concentration of doxycicline for metalloproteinase-2 was 138 microM (r2 = 0.82). CONCLUSIONS: Internal mammary arteries and saphenous veins have different metalloproteinase activities, that are inhibited by doxycicline. PMID- 9336065 TI - [Clinical effectiveness of and tolerance to ramipril in the treatment of essential arterial hypertension phase 1 and 2. Results of a multicenter study]. AB - BACKGROUND: In the last two decades, angiotensin converting enzyme inhibitors have become first line medications in the treatment of hypertension. AIM: To assess the tolerance and effectiveness of ramipril as monotherapy in the treatment of essential hypertension. PATIENTS AND METHODS: One hundred eighty eight hypertensive patients, aged 21 to 80 years-old, coming from 4 Chilean hospitals were studied. Using an open non controlled design; they were treated with placebo for two weeks and with the active drug during eight weeks, in initial doses of 2.5 mg/day that were adjusted to 5 mg/day if diastolic blood pressure did not drop below 90 mm Hg or if its reduction was less than 10 mm Hg. RESULTS: During the active drug treatment period, systolic blood pressure decreased from 164.8 +/- 7.2 to 147.3 +/- 4.8 mm Hg. Diastolic blood pressure dropped from 102.3 +/- 3.1 to 87.8 +/- 3.0 mm Hg. Seventy percent of patients achieved a diastolic blood pressure of less than 90 mm Hg, 56.9% with 2.5 mg/day and 13.8% with 5 mg/day. Dizziness, cough and headache were the main adverse reactions, observed in 3.7, 3.2 and 2.1% of patients respectively. Adherence to treatment was 98%. There were no changes in laboratory values during the treatment period. CONCLUSIONS: Ramipril is effective and well tolerated in the treatment of essential hypertension. PMID- 9336064 TI - [Markers of hepatic fibrogenesis in alcoholic patients]. AB - BACKGROUND: An elevation of serologic markers of hepatic fibrogenesis has been reported in liver diseases of different etiologies. Among these, the N-terminal type III procollagen (P-III-P) and the P1 proteolytic fragment of laminin (P1 laminin) increase in alcoholic liver damage, in proportion to the progression of this condition. AIM: To study serum levels of P-III-P and P1 laminin in asymptomatic alcoholics with and without liver damage and decompensated alcoholic cirrhotics, compared to normal controls. METHODS: Serum P-III-P and laminin levels were measured in asymptomatic alcoholics during detoxification treatment. Liver biopsies were obtained, in order to detect liver damage, which was graded with a numeric score, considering values over 6 as severe damage. Serum fibrogenesis markers were also measured in a group of decompensated alcoholic cirrhotics. RESULTS: P-III-P levels were significantly higher in cirrhotic patients compared to alcoholics with or without liver damage and to normal controls. Laminin was not different between groups. P-III-P did not correlate with histologic score in asymptomatic patients. CONCLUSIONS: In this study P-III P and P1 laminin were not usefull discriminators of severe liver damage among asymptomatic alcoholics; their levels were found to rise significantly only when liver disease has become clinically evident. PMID- 9336066 TI - [Growth hormone deficiency in patients with chronic heart failure]. AB - BACKGROUND: Experimental and preliminary clinical data in patients with dilated cardiomyopathy show that growth hormone has a positive inotropic effect and contributes to peripheral vasodilatation. However, there is little information about the activity of growth hormone-IGF-1 axis in patients with chronic heart failure. AIM: To measure growth hormone and IGF-1 levels in patients with chronic heart failure. PATIENTS AND METHODS: Nine patients, aged 49 to 76 years old, 7 male, were studied. Seven had an idiopathic dilated cardiomyopathy and 2 a coronary heart disease. All had a stable cardiac failure, in functional capacity II or III and were receiving digoxin, furosemide and potassium supplements. Thyroid hormone levels, basal and exercise growth hormone and IGF-1 levels were measured and compared with reference values for American populations. Left ventricular ejection fraction was measured with an isotopic technique and nutritional status using anthropometry and indirect calorimetry. RESULTS: Anthropometric measures, basal and post-prandial oxygen consumption were within normal limits. Thyroid hormone levels were normal. During maximal exercise, growth hormone levels were 2.56 +/- 4.1 ng/ml and IGF-1 levels were 0.56 +/- 0.61 mU/ml. These values were significantly lower than expected for age and sex. CONCLUSIONS: These patients with chronic cardiac failure have lower than normal growth hormone and IGF-1 levels. PMID- 9336067 TI - [Assessment of neurotoxic effects of methyl bromide in exposed workers]. AB - BACKGROUND: Methyl bromide is an aliphatic hydrocarbon derivative used as a pesticide that causes skin, kidney, respiratory, liver and neurological damage. AIM: To assess the neurological and psychiatric damage caused by methyl bromide in exposed workers of seed and fruit export industries in a rural area near Santiago. SUBJECTS AND METHODS: We studied prospectively 15 male middle age workers before and after a fumigation period with methyl bromide, that lasted two to four weeks. According to the initial assessment, 5 of these subjects had a chronic exposure to the chemical. As controls, 10 non exposed workers matched for age, sex and working conditions were studied in two occasions. The evaluation included the WHO Neuro Behavior Core Test Battery, dynamometric and vibrator assessment of peripheral nerve function, the Nothingham test for psychological functioning and Titmus test for visual acuity. Methyl bromide levels were measured in blood and urine. RESULTS: Blood methyl bromide levels increased from 13.3 to 30 mg/dl after exposure. Symptoms that appeared with a higher frequency in exposed workers were insomnia, headache, paresthesiae, mood changes and loss of memory and concentration. In these subjects, the threshold for the Vibraton test increased from 2.4 to 2.85 sec, dynamometry showed a strength reduction in the right side from 51.4 to 47.2 kg and there was an increase in the score for negative auto-perception in the Nothingham test from 11.2 to 13.6. No deterioration in these tests were observed in unexposed workers. CONCLUSIONS: Acute and chronic methyl bromide exposure causes important psychological and neurological derangement. PMID- 9336068 TI - [Surgical treatment of diffuse hyperthyroid goiter. Experience at a hospital in Valdivia, Chile]. AB - BACKGROUND: The choice of medical, radiation or surgical therapy for hyperthyroid diffuse goiter is still empirical. AIM: To report a retrospective analysis of the surgical treatment of hyperthyroid goiter in a Regional Hospital in Chile. PATIENTS AND METHODS: The charts of 64 patients, 54 female, aged between 15 and 57 years old, operated between 1985 and 1995 were analyzed. RESULTS: The indication for surgical treatment was failure of medical treatment in 59 patients and a big goiter causing mechanical compression in 3 patients. A subtotal thyroidectomy was done after an abbreviated surgical preparation. The mean weight of the resected glands was 65.9 g. Four patients had transient hypocalcemia and 4 had surgical wound seromas. After a mean follow up of 31 months, 77% of patients remain euthyroid, hyperthyroidism relapsed in 13.1% and 10% became hypothyroid. CONCLUSIONS: Surgical treatment of hyperthyroid goiter is safe but the percentage of hyperfunction relapse is high. PMID- 9336069 TI - [Adrenal macrotumor diagnosed by computed tomography. Clinical experience of 33 cases]. AB - BACKGROUND: Adrenal tumors of more than 6 cm diameter, also called macrotumors, cause diagnostic and therapeutic problems. AIM: To perform an analysis of adrenal macrotumors diagnosed by CAT scan between 1984 and 1995. PATIENTS AND METHODS: Thirty three patients aged 9 to 86 years old, 15 female, with an adrenal macrotumor diagnosed by CAT scan, were analyzed. RESULTS: Thirty percent of tumors were functioning (70% secreted cathecolamines and 30% cortisol). Eighty two percent had an adrenal localization and 18% were para-adrenal. Thirty four percent were malignant. These tumors were mostly non functioning, 70% occurred in men and 67% were metastatic. CONCLUSIONS: Most adrenal macrotumors in this series were non functioning and 36% were malignant. PMID- 9336070 TI - [Hydrostatic reduction of intestinal intussusception in children. Experience in 43 cases]. AB - BACKGROUND: Intestinal intussusception, a medical emergency, is more commonly idiopathic and ileocolic and occurs with higher frequency in children aged 6 months to 2 years. Barium enema confirms the diagnosis and allows its hydrostatic reduction, that is the treatment of choice of this condition. AIM: To report our experience with hydrostatic reduction of intestinal intussusception in children. PATIENTS AND METHODS: Hydrostatic reduction was attempted in 43 children with intestinal intussusception: 20 male, aged 2 to 48 months, that consulted at the Clinical Hospital of the Catholic University in Santiago. RESULTS: Hydrostatic reduction was successful in 33 children (77%) that were discharged from the hospital 24 to 96 hours after the procedure. A partial reduction was achieved in 10 patients (23%) who required surgery and were discharged from the hospital 5 to 8 days after the procedure. CONCLUSIONS: Our results are similar to those reported abroad and allow the recommendation of hydrostatic reduction as the treatment of choice for intestinal intussusception. PMID- 9336072 TI - [Breast hydatid cyst. A case report]. AB - We report the case of a 63 years old woman with a left mammary hydatid cyst. The patient presented with a left painless mammary mass of four months evolution. Mammography showed a dense image of 9 cm phi in the left breast. Ultrasound showed a solid and heterogeneous lesion. The tumor was excised under general anesthesia. The initial macroscopic inspection showed a tumor with a well defined fibrous capsule filled with a grayish material that contained membrane debris. The pathological study confirmed the diagnosis of a hydatid cyst. PMID- 9336071 TI - [Metalloproteinases in vascular wall remodelling]. AB - Metalloproteinases (MTP) are enzymes that degrade the extracellular matrix, mainly collagen tissue. Normally these enzymes are expressed in vascular walls as proenzyme together with inhibitors of the active enzymes. By effect of different cytokines, produced by an inflammatory process in the vascular wall, these proenzymes are activated to an extent that surpasses the action of the inhibitors and degrade collagen. This action may partly explain the rupture of atherosclerotic plaques ("vulnerability") and also the remodelling of the vessel wall with "compensatory enlargement" of the vessel (increase in the outer size of the vessel) that allows the plaques to develop inside the arterial wall without protruding into the vessel lumen for many years. The occlusion of saphenous vein in aortocoronary bypass grafts is due to fibromuscular proliferation and atheroma development and therefore the participation of MTP in the occlusion of these vessels is a reasonable hypothesis. However, the structural features of saphenous vein bypass grafts are different from those of atheroma in native coronary arteries. Mainly the compensatory enlargement of the vessels does not occur because of intense fibrous tissue development including the adventitia and therefore the new tissue in the wall is forced to protrude into the vessel lumen. The reason for this difference in the vessel wall remodeling is not clear and the article by Grez et al in this issue of this Journal is an starting and promising study in this regard. PMID- 9336073 TI - [Ethics code of the Chilean Biological Society]. AB - The Chilean Biological Society has approved an ethics code for researchers, elaborated by its Ethic Committee. The text, with 16 articles, undertakes the main ethical problems that researchers must solve, such as institutional, professional or societal ethics, scientific fraud, breaches in collaborative work, relationships between researchers, participation in juries and committees, ethical breaches in scientific publications, scientific responsibility and punishments. This code declares its respect and valorization of all life forms and adheres to international biomedical ethical codes. It declares that all knowledge, created or obtained by researchers is mankind's heritage. PMID- 9336074 TI - [Emergent diseases: old and new diseases. Etiological and climatic aspects. Socioeconomic and cultural influences]. AB - During recent years the world has experienced the reemergence of old communicable diseases and the emergence of new ones caused by novel pathogens such as the HIV virus and Borrelia burgdorferi. The problem consists mostly in the reemergence of old diseases but specially in industrialized countries new pathogens have also been described. Although the emergence of these infections in rare instances is due to genetic changes of pathogens to more virulent forms, most commonly they are due to changes in the environment and the host. Rapidly deteriorating living standards, disintegration of sanitation and public health infrastructure, cultural changes, migration and variations in behavior are some of the factors involved in the worldwide increase of infectious diseases. The degradation of natural habitats including forests and marine niches accompanied by climatic changes, are also playing an increasing role in the detrimental evolution of these diseases. The global emergence of these diseases calls into question the doctrine of epidemiological transition and directs us to scrutinize the paradigm that bases the prevention of these diseases solely on vaccination. The current situation also highlights the limitations of classical epidemiology in dealing with unexpected problems, and strongly suggests that this discipline should incorporate into its analysis findings from other fields, including ecological, climatological, and economical information. As most of the negative social and economical developments that impinge on the detrimental evolution of these diseases are increasing world-wide, it can be predicted that the problems posed by these infections will continue and perhaps worsen in the near future. PMID- 9336075 TI - [Late effects of head trauma]. AB - The recuperation after a head trauma is divided in three phases. Awakening from coma is the initial phase that normally occurs at the intensive care unit. The period of hospitalization when the threatens of death and neurological instability disappear is the intermediate phase. The post concussional syndrome occurs in the late phase, when the patients have been discharged from the hospital. This syndrome, despite the absence of abnormalities in the clinical examination, causes an assortment of ailments that preclude normal activities. Organic neurological sequelae such as hemiplegia, speech disorders, cranial nerve lesions and mental disorders take also place in this phase. An adequate rehabilitation plan for the patient and his family must take into account all these issues. PMID- 9336076 TI - [Renal complications in HIV-1 infection]. AB - Renal involvement in AIDS may be specific or unspecific. Unspecific lesions, the most common, are usually an acute tubular necrosis produced by hemodynamic, infectious or electrolytic alterations that lead to an acute renal failure or drug nephrotoxicity. Specific lesions are segmental and focal hyalinosis, immune complex glomerulonephritis and thrombotic microangiopathy. Focal and segmental hyalinosis is observed almost exclusively in black people and produces a rapidly progressive renal failure. Lesions are a consequence of HIV stimulation of TGF beta in mesangial cells. Immune complex glomerulonephritis, formed by HIV antigens and anti HIV antibodies, is observed in white and black people. The glomerular lesion in this condition is less severe than in the former. Thrombotic microangiopathy could be a consequence of a pathogenic effect of the virus over glomerular capillaries and arterioles. It is clinically expressed as a hemolytic uremic syndrome. This paper reports briefly the renal pathological study of 46 patients infected with HIV-1, seen at the Nephrology Service of the Bichat Hospital in Paris. PMID- 9336077 TI - [Disability in the private pension system in Chile]. AB - BACKGROUND: The degree of disability of workers ascribed to the private allowances system in Chile, is judged by Medical Commissions that apply norms that establish percentages of incapacity, without considering prognosis. AIM: To communicate the causes of disability among Chilean workers ascribed to the private allowances system, their mortality and to correlate the causes of death with diagnoses. SUBJECTS AND METHODS: We analyzed 13,456 consecutive cases judged between August 1990 and April 1992. Mortality was registered up to 12 months after judgment. RESULTS: Total incapacity was determined in 4,158 cases (30.9%), partial incapacity in 1,340 (9.9%) and minor incapacity in 7,958 (59.1%). Osteoarticular diseases were the main cause of disability in 4,460 patients (33.1%) and 77.8% of patients with malignant tumors were considered as having total incapacity. Mortality was 17% among subjects with total incapacity, 1.5% among those with partial disability and 1% among those with minor disability. The cause of death was related to the main disabling disease in 94% of subjects with total incapacity and 66.6% of those with partial incapacity. CONCLUSIONS: Osteoarticular diseases are the main cause of inability among workers ascribed to a private pension system. PMID- 9336079 TI - [When and by whom was the Clinical Hospital left under the management and administration of the Dean of the University of Chile School of Medicine?)]. PMID- 9336078 TI - [Gastric cancer: a unique etiological entity?]. AB - Based on epidemiological, clinical and pathological data, the existence of two etiologic entities, for what is generically considered as gastric carcinoma, is postulated. Environmental factors would be more important in the etiology of the differentiated type of gastric cancer and genetic factors would have a relevant role in the etiology of the undifferentiated type. Geographic, ethnic, environmental and genetic factors participate in the epidemiology of gastric cancer. This review emphasizes the evidences supporting the role of genetic factors in the etiology of undifferentiated gastric cancer. The importance of looking for a shared phenotype among patients with this type of cancer is underscored. PMID- 9336080 TI - A proposed heuristic for communicating heritability estimates to the general public, with obesity as an example. AB - It is well established that many continuously distributed traits have a heritable component. However, it is often difficult to communicate to the general public the meaning of quantitative estimates of heritability. To address this problem, the present paper introduces a heuristic for communicating heritability to nonscientific audiences. This heuristic involves adopting an extremely simplified model of inheritance and artificially (and somewhat arbitrarily) defining a cutoffs of "low environmental risk" and "affectation status." Using body weight and obesity as an example, we present a table which gives estimates of the proportion of obese persons who are "genetically obese" assuming varying levels of "environmental risk" for obesity and relative body weight scores for defining obesity. The resulting statistic may prove useful for lay audiences in understanding a heritability estimate. PMID- 9336081 TI - Comorbidity of mathematics and reading deficits: evidence for a genetic etiology. AB - In order to assess the genetic etiology of the comorbidity of reading and mathematics difficulties, data were ascertained from two samples: (1) 102 identical and 77 same-sex fraternal twin pairs in which at least one member of each pair is reading disabled and (2) 42 identical and 23 same-sex fraternal twin pairs in which at least one member is math disabled. Composite reading and mathematics performance data from each sample were fitted to the basic multiple regression model for the analysis of selected twin data and its bivariate extension. Resulting estimates of bivariate heritability and the genetic correlation between the reading and the mathematics performance measures suggest that the comorbidity between mathematics and reading difficulties is due in part to genetic influences. PMID- 9336082 TI - The genetic correlation between impulsivity and sensation seeking traits. AB - A number of studies have demonstrated associations between sensation seeking traits and measures of impulsivity. This study examined contributions to the observed correlations between imupulsivity and sensation seeking traits. Fifty seven pairs of identical and 49 pairs of fraternal twins who were reared apart and 90 individuals who also participated in the Minnesota Study of Twins Reared Apart completed the Control scale of the Multidimensional Personality Questionnaire (MPQ; Tellegen, 1982) and the four subscales of the Sensation Seeking Scale (SSS; Zuckerman, 1979). Consistent with previous studies, the Control scale was significantly correlated with the SSS. A Cholesky decomposition of the data indicated that the phenotypic correlations between the Control scale and the four subscales of the SSS were mediated almost entirely by genetic factors. In the final reduced model the proportion of the genetic variance of the Control scale in common with the SSS was estimated as 55%, and the rest of the genetic variance (45%) was attributed to the genetic variance specific to the Control scale. The results emphasize the importance of common biological mechanisms underlying associations between impulsivity and sensation seeking traits. PMID- 9336083 TI - Genomic imprinting and audiogenic seizures in mice. AB - Audiogenic seizure (AGS) susceptibility in mice is a multifactorial behavioral disorder that involves severe generalized convulsions in response to loud, high frequency sound. The inheritance of AGS susceptibility was examined in crosses between AGS-susceptible DBA/2J (D2) mice and epilepsy-prone (EP) mice. The EP mice were selected for high AGS susceptibility in a BALB/c-derived line. The AGS phenotype was similar in the EP and D2 mice at 30 days of age. The frequency of generalized clonic-tonic AGS was high in both the D2 and the EP mice (53 and 83%, respectively) but was low in the reciprocal EPD2F1 and D2EPF1 hybrids (14 and 19%, respectively). In the backcross to the EP parent, no significant associations were found between AGS susceptibility and microsatellite markers linked to Asp1 or Asp2, AGS genes located on Chromosomes 12 and 4, respectively. Significant associations were found for markers linked to Asp3, which is located in the proximal region of Chromosome 7. The influence of Asp3 on AGS susceptibility was seen in the EP x EPD2F1 backcross but not in the reciprocal EPD2F1 x EP backcross, suggesting that Asp3 expression is influenced by genomic imprinting. A model is proposed where genomic imprinting represses the maternal Asp3 allele, providing an influence largely from the paternal allele. PMID- 9336084 TI - No evidence for a Y chromosomal effect on alternative behavioral strategies in mice. AB - This study takes the first step toward testing a Y chromosomal effect on both aggression and thermoregulatory nest-building behavior in mouse lines either bidirectionally selected for short (SAL) and long (LAL) attack latency or high (HIGH) and low (LOW) nest-building behavior. Using reciprocal crosses between SAL and LAL, and between HIGH and LOW, we found no indications for Y chromosomal effects on thermoregulatory nest-building behavior. As for aggression, we confirmed earlier studies on SAL and LAL, i.e., the origin of the Y chromosome influences attack latency, i.e., aggression. However, we did not find indications for a Y chromosomal effect on aggression in the HIGH and LOW lines. Since aggression and nest-building behavior have been shown to be characteristic parameters of two fundamentally different behavioral strategies, the present data underline the improbability of Y chromosomal genes underlying the genetic architecture of alternative behavioral strategies. PMID- 9336085 TI - Ovulation and the suppression of mating in Drosophila melanogaster females: behavioral basis. AB - Virgin females of Drosophila melanogaster that are ectopically expressing the sex peptide gene show a high level of ovulation and are unreceptive to males. However, if they are genetically deprived of eggs, receptivity is considerably restored (Fuyama, 1995). These females, whether they have eggs or not, extrude their ovipositors toward courting males as frequently as do fertilized females. However, this rejection behavior was ineffective in suppressing male courtship. Of females with eggs, about half of them could suppress male courtship. Females lacking eggs could not suppress male courtship and continued to elicit vigorous courtship. This difference seems to account for the increased mating frequency in sterilized females. Courtship behavior by mutant males defective in olfaction or learning suggested that females are capable of repelling males by emitting a volatile pheromone(s) with an inhibitory effect on male courtship. PMID- 9336086 TI - Correlated responses to selection for developmental period in Bactrocera cucurbitae (Diptera: Tephritidae): time of mating and daily activity rhythms. AB - Comparisons of "time of mating" (the time copulation begins) between lines selected for short and long developmental periods have been made in the melon fly, Bactrocera cucurbitae. These comparisons showed that longer development periods were associated with later initiation of mating. Crosses were also made between selected lines to ascertain the genetic basis of developmental period and time of mating. Comparisons of daily activity rhythms for four types of behavior (locomotion, preening, feeding, and wing vibration) between the selected lines showed the following; (1) locomotion and preening occurred in daytime for the short developmental period lines, versus mainly at night for the long developmental period lines; (2) feeding behavior occurred in daytime for both the short and the long developmental period lines; and (3) wing vibration, a component of courtship behavior of males, occurred at dusk for the short developmental period lines and at night for the long developmental period lines. PMID- 9336087 TI - Back to basics in the fight against tobacco. PMID- 9336088 TI - Smoking in Ontario, 1991 to 1996. AB - Surveys by the Addiction Research Foundation of Ontario have produced annual estimates on smoking prevalence since 1991. This report describes the three series of telephone surveys from which these data are drawn as well as future plans to monitor tobacco use in Ontario. In addition to provision of updated descriptive results, the methodology and limitations of the data are discussed. Prevalence data for 1996 are presented from the Ontario Drug Monitor, a telephone survey of Ontario adults (n = 2721). The overall prevalence of smoking in Ontario was 27% (95% confidence interval: 25% to 29%); 23% smoked daily (95% confidence interval: 21% to 25%). There is no evidence of any decline in the prevalence of smoking since 1991, and no sex differences were found in smoking prevalence. Future reports will update trend data and provide robust regional estimates. PMID- 9336089 TI - Canadian immunization cuts will cause child deaths. PMID- 9336090 TI - Tobacco industry campaign contributions in Ontario, 1990-95. PMID- 9336091 TI - A Canadian tertiary care centre study of maternal and umbilical cord cotinine levels as markers of smoking during pregnancy: relationship to neonatal effects. AB - This study describes the prevalence of smoking among 3,220 pregnant women. Maternal and umbilical cord cotinine levels were compared with the women's self reported cigarette consumption, infant birth weight and antepartum and perinatal complications. Of the women who reported themselves as being active smokers (23%), 76% had a partner who smoked, and 38% reported exposure to environmental smoke in the workplace. Only 15% of nonsmokers had a partner who smoked, and 13% reported workplace exposure. The mean number of cigarettes/day was 20.5 (95% CI 19.6-21.4). The relative risk of having a small-for-gestational-age infant was significantly higher in smokers for mothers of both preterm (34-36 wks, RR = 3.38, 95% CI 1.25-9.16) and term babies (> or = 37 wks, RR = 2.04, 95% CI 1.58 2.63). Mean infant birth weight was 207 g lighter in the infants of smokers (p < 0.001) and was inversely correlated to maternal serum cotinine level. Birth weight dropped by 0.99 g for every 1 ug/L increase in cotinine (r = -0.19, p < 0.01). PMID- 9336092 TI - Factors influencing the duration of breastfeeding in the Sudbury region. AB - OBJECTIVE: To determine the duration of breastfeeding in the Sudbury Region and to identify the reasons why mothers wean before the Canadian Paediatric Society's recommended six month period. METHODS: Questionnaires addressing factors that influence the duration of breastfeeding were mailed to 350 breastfeeding mothers at one/two weeks, three months and six months after their postpartum discharge from hospital. RESULTS: Forty percent of mothers breastfed for the recommended six month period. Reported factors positively influencing longer durations of breastfeeding were higher education, higher family income, parity, previous breastfeeding experience, decision to breastfeed before the child was born and late introduction to solids. Reasons for weaning included perceived insufficient milk supply, fatigue, breast problems and return to work. Mothers' top three choices of services were home visits, telephone hot line and television programs. CONCLUSIONS: The duration of breastfeeding in the Sudbury Region is lower than the provincial average. Several modifiable factors associated with duration of breastfeeding were identified. PMID- 9336093 TI - Using a breastfeeding prevalence survey to identify a population for targeted programs. AB - Often, efforts to improve overall population health require identifying and targeting programs to specific high-risk populations. Breastfeeding is an example. In order to determine initiation and duration rates among various groups in the City of Toronto, a random sample of 434 mothers with infants at four months of age was surveyed to determine the prevalence of breastfeeding and major impacts on its duration. The study found that, overall, 83% of mothers initiated breastfeeding at birth. The greatest rate of decline occurred during the first month. At four months postpartum, 57% of mothers continued to breastfeed, including 35% who were exclusively breastfeeding and 22% who were supplementing breast milk with formula. Breastfeeding duration was related to a number of factors, including information and support, parity, education, use of formula supplements and country of birth. Specific groups are identified for targeted programs, and a number of strategies are proposed. PMID- 9336094 TI - Public health in Canada: a comparative study of six provinces. AB - PURPOSE: To describe the public health systems and their projected futures in six provinces in the context of two developments: 1) the emerging discourse on population health and 2) the trend toward regionalization of the health care system. METHODS: Telephone interviews with key informants and key document review. RESULTS AND CONCLUSIONS: Communicable disease control and health protection are currently the "core businesses" of public health; the population health discourse has not resulted in mandated programming. The reality is a retrenchment of public health scope during a time that should be considered conducive to expansion. Only Ontario has not regionalized its health care system, although public health is already delivered regionally. Alberta, Saskatchewan and Manitoba have either evolved or are evolving toward an integrated health system. There were concerns about the potential impact on public health identity and funding of this "vertical integration". Regionalization of public health may result in units that are too small to support adequate local expertise and may jeopardize development and enforcement of province-wide programs. PMID- 9336095 TI - Mathematical models of disease transmission: a precious tool for the study of sexually transmitted diseases. AB - This paper is an introduction to the mathematical epidemiology of sexually transmitted diseases (STDs) and its application to public health. After a brief introduction to transmission dynamics models, the construction of a deterministic compartmental mathematical model of HIV transmission in a population is described. As a background to STD transmission dynamics, basic reproductive rate, intergroup mixing, rate of partner change, and duration of infectivity are discussed. Use of the models illustrates the effect of sexual mixing (proportionate to highly assortative), of preventive intervention campaigns, and of HIV-chlamydia interaction on HIV prevalence in the different population groups. In particular, planned prevention campaigns can benefit the targeted intervention group but surprisingly can be disadvantageous for the general population. Through examples, mathematical models are shown to be helpful in our understanding of disease transmission, in interpretation of observed trends, in planning of prevention strategies, and in guiding data collection. PMID- 9336096 TI - Prevalence of chlamydial infection and frequency of risk behaviours for STDs and HIV infection among adolescents in public juvenile facilities in the province of Quebec. AB - This study aimed to determine the prevalence of chlamydial infection in adolescents entering public juvenile facilities in the province of Quebec and the frequency of their risk behaviours for STDs and HIV infection. Adolescents were asked to complete an anonymous self-administered questionnaire and to be screened for Chlamydia trachomatis. Of 731 sexually active adolescents, 62% agreed to be tested. The overall prevalence rate was 7.7% (95% confidence interval: 5.1% 10.3%). The prevalence was significantly higher in female than in male teenagers: 12% vs 3% (p < 0.001, Fisher's exact test). These results suggest that screening for chlamydial infection should be offered to all sexually active female teenagers admitted into juvenile facilities. For males, selective screening taking into account sexual history would be a more realistic approach. The frequency of high risk activities for STDs and HIV infection points to the importance of specific prevention programs for this population. PMID- 9336097 TI - Effectiveness of recorded messages to communicate the risk of acquiring hantavirus pulmonary syndrome. AB - OBJECTIVES: To determine the effectiveness of a recorded information line in communicating health risk during the emergence of a new disease, hantavirus pulmonary syndrome (HPS), in the Edmonton area and to study the accuracy of recall of information about hantavirus among the general public. METHODS: A random telephone survey of residents five months after a death from HPS had occurred. RESULTS: The number of residents who received their information from the recorded line was quite low (approximately 2%), and more people remembered receiving their information through the news media, particularly television (74%) and newspaper (57%). CONCLUSIONS: An information line by itself will not communicate risk effectively during an outbreak or other emergent health situation. However, an information line used in conjunction with news media proved effective in providing ongoing, accurate information and allaying public fears in a low-risk situation. PMID- 9336098 TI - Responding to reported clusters of common diseases: the case of multiple sclerosis. AB - Reports of disease clustering are becoming ever more common, and there is increasing pressure on public health agencies to respond rapidly and appropriately to these reports. We investigated a cluster of five cases of MS occurring in a small multidisciplinary hospital department. Data were collected by a variety of methods, including measurement and description of the workplace, completion of survey instruments by department staff, and construction of case histories of persons with MS. The results indicated that the department comprised a high-risk population and that only one case of MS could have any possible etiologic significance. Investigators should consider a number of factors when evaluating disease clusters, including the accuracy of diagnosis, the induction period and cause of the disease, and possible biases in the population at risk. Additionally, boundaries should not encircle the cases that led to identification of the cluster and should reflect environmental significance. PMID- 9336099 TI - Correlates of condom use in the young adult population in Ontario. AB - OBJECTIVES AND METHODS: Data from the Ontario Health Survey were used to identify sociodemographic, lifestyle and sexual history characteristics associated with the use of condoms for protection against sexually transmitted diseases (STDs) in randomly selected adults between the ages of 16 and 44 years who had had two or more sexual partners in the 12 months before the survey (n = 2,699). RESULTS: Forty-two percent reported not having used condoms for protection against STDs. Those most likely to use condoms were 16 to 24 years of age, males, students, non binge-drinkers, urban residents, and those at higher risk for HIV/AIDS. Of those who used condoms, 68% did not use them consistently. Individuals most likely to always use condoms were 16 to 24 years of age, males, students, non-binge drinkers, and those with secondary school education. Age, gender, occupational activity, and non-binge-drinking were common correlates of both condom use and consistent use. CONCLUSIONS: Public health messages should be focused on people with multiple sex partners who are not using condoms for STD protection, including rural residents, those with high levels of education, and those over 34 years of age. PMID- 9336100 TI - The natural history of amphibian skin secretions, their normal functioning and potential medical applications. AB - Amphibians occupy a wide range of habitat types from arid deserts to deep freshwater lakes; they may spend most of their life underground or high in cloud forest canopy. Some are found north of the Arctic Circle and can tolerate freezing conditions, while others have evolved a range of adaptations to avoid desiccation in some of the hotter areas of the world. The skin plays key roles in the everyday survival of amphibians and their ability to exploit a wide range of habitats and ecological conditions. The normal functions of the skin are surveyed and Eisner's biorational approach to chemical prospecting--seeking clues from an animal's behaviour and its interactions with its environment to reveal the presence of chemical compounds with potential medical or veterinary applications- is applied to amphibians. The biology and natural history of amphibian skin, its glands and their secretions are briefly reviewed. Four categories of compounds are found in the granular or poison glands, these are: biogenic amines, bufodienolides (bufogenins), alkaloids and steroids, peptides and proteins. Toads, particularly members of the genus Bufo, are identified as a particularly convenient and useful source of granular gland secretions. The potential medical pharmaceutical significance of products derived from amphibian skin secretions is discussed. The need for a humane approach to this work is noted. PMID- 9336102 TI - Surface electromyography and mouse use position. AB - This study examines muscle tension and subjective muscle tension awareness while using a computer mouse positioned to the right of a standard computer keyboard and a centrally positioned trackball. Seventeen volunteer subjects experienced in mouse and trackball use were seated at an ergonomically adjusted workstation. Surface electromyography (sEMG) and subjective muscle tension levels were recorded from four muscle groups (left sternocleidomastoid/scalene, right upper trapezius, right posterior deltoid, and right lower trapezius/rhomboids) during 1 min trials with subjects resting with hands in their lap, while using a trackball below the centre of the keyboard, and while using a mouse immediately to the right of a 101-key keyboard. All subjects showed significantly higher mean sEMG activity recorded from the right upper trapezius, right posterior deltoid, and right lower trapezius/rhomboids during mouse use to the right of a standard keyboard (arm abducted) compared to using a trackball positioned centrally, (p < 0.001). sEMG levels remained elevated during the entire trial period of right side mouse use without evidence of microbreaks (< 1 s epochs of low sEMG activity). sEMG activity from the left sternocleidomastoid/scalene muscles showed no significant change from baseline in any condition. Subjective reports of muscle tension did not correlate with sEMG activity. The authors predict that there will be an overall increase in reports of upper extremity musculoskeletal disorders (UEMSD) and computer related disorders (CRD) when people abduct their arms in order to reach a mouse positioned to the side of standard width, or wider keyboards. Discussed are the applications of sEMG for evaluation of computer keyboard and pointing device use, appropriate ergonomic equipment design, and a methodology for improving muscle awareness, strengthening, relaxation, and workstyle practices to promote healthier computing. PMID- 9336103 TI - A fuzzy linguistic model for the prediction of carpal tunnel syndrome risks in an occupational environment. AB - This research presents the development and evaluation of a fuzzy linguistic model designated to predict the risk of carpal tunnel syndrome (CTS) in an occupational setting. CTS has become one of the largest problems facing ergonomists and the medical community because it is developing in epidemic proportions within the occupational environment. In addition, practitioners are interested in identifying accurate methods for evaluating the risk of CTS in an occupational setting. It is hypothesized that many factors impact an individual's likelihood of developing CTS and the eventual development of CTS. This disparity in the occurrence of CTS for workers with similar backgrounds and work activities has confused researchers and has been a stumbling block in the development of a model for widespread use in evaluating the development of CTS. Thus this research is an attempt to develop a method that can be used to predict the likelihood of CTS risk in a variety of environments. The intent is that this model will be applied eventually in an occupational setting, thus model development was focused on a method that provided a usable interface and the desired system inputs can also be obtained without the benefit of a medical practitioner. The methodology involves knowledge acquisition to identify and categorize a holistic set of risk factors that include task-related, personal, and organizational categories. The determination of relative factor importance was accomplished using analytic hierarchy processing (AHP) analysis. Finally a mathematical representation of the CTS risk was accomplished by utilizing fuzzy set theory in order to quantify linguistic input parameters. An evaluation of the model including determination of sensitivity and specificity is conducted and the results of the model indicate that the results are fairly accurate and this method has the potential for widespread use. A significant aspect of this research is the comparison of this technique to other methods for assessing presence of CTS. The results of this evaluation technique are compared with more traditional methods for assessing the presence of CTS. PMID- 9336104 TI - The effect of keyboard keyswitch make force on applied force and finger flexor muscle activity. AB - The design of the force-displacement characteristics or 'feel' of keyboard keyswitches has been guided by preference and performance data; there has been very little information on how switch 'feel' alters muscle activity or applied force. This is a laboratory-based repeated measures design experiment to evaluate the effect of computer keyboard keyswitch design on applied finger force and muscle activity during a typing task. Ten experienced typists typed on three keyboards which differed in keyswitch make force (0.34, 0.47 and 1.02 N) while applied fingertip force and finger flexor electromyograms were recorded. The keyboard testing order was randomized and subjects typed on each keyboard for three trials, while data was collected for a minimum of 80 keystrokes per trial. No differences in applied fingertip force or finger flexor EMG were observed during typing on keyboards with switch make force of 0.34 or 0.47 N. However, applied fingertip force increased by approximately 40% (p < 0.05) and EMG activity increased by approximately 20% (p < 0.05) when the keyswitch make force was increased from 0.47 to 1.02 N. These results suggest that, in order to minimize the biomechanical loads to forearm tendons and muscles of keyboard users, keyswitches with a make force of 0.47 N or less should be considered over switches with a make force of 1.02 N. PMID- 9336105 TI - Subjective perceptual methods for comparing backpacks. AB - Subjective perceptual methods may provide useful information about small differences in backpack design when physiological and biomechanical comparisons are ineffective. This study used two subjective perceptual methods, category ratio scale (CRS) ratings of perceived discomfort and written questionnaires for comparing two types of leisure backpack. CRS ratings of perceived discomfort for each of 24 body regions after 30 min of uphill (15% grade) treadmill walking at 3 km h-1 in 10 males, failed to distinguish between a New Zealand designed backpack (Pack A) and a British designed backpack (Pack B), each weighted to 20 kg. A simple pre- and post-walking written questionnaire using either a visual analogue linear scale or free-format responses indicated that more subjects found Pack A easier to adjust but that it had less comfortable shoulder and waist straps. It was considered to be more comfortable with regard to balance and posture and for shoulder, back and leg muscular tension. Pack B was initially more comfortable but required more lumbar support. Pack B was considered more comfortable for waist and shoulder pressure only. Overall preference was for Pack A (seven subjects) rather than Pack B (three subjects). In conclusion, in this study a questionnaire approach was found to be more useful than CRS ratings of perceived discomfort and the New Zealand designed backpack was preferred. PMID- 9336106 TI - Perception and visualization of human posture information for computer-aided ergonomic analysis. AB - This article reports three experiments that examined the effects of photographic method, computerized visualization scheme, and posture complexity on posture perception and specification for computer-aided ergonomic analysis. The subjects were presented with photographs of working postures, and were required to manipulate human forms generated by an ergonomics software program to match the postures in the photographs. The first experiment showed a clear advantage of using a three-dimensional (3-D) human form graphic with two photographs when complex, asymmetric postures were analysed. However, the use of a 3-D human graphic display and two photographs jeopardized the subjects' posture specification performance when simple, symmetric postures were analysed. The results of the second and the third experiment demonstrated the importance of achieving congruency between photographic and computer display perspectives in improving posture specification performance. Implications for ergonomic job analysis and ergonomics software design are discussed. PMID- 9336107 TI - Effects of arm support on shoulder and arm muscle activity during sedentary work. AB - The aim of this study was to evaluate different arm supports by comparing the activity of shoulder and arm muscles during various work tasks, with and without the lower arm supported. Twelve female subjects, aged between 23 and 37 years, were asked to perform three types of tasks: typing, simulated assembly work (in two different positions), and pipetting. The supports used were: fixed arm support (FIX), horizontal movable arm support (HOR), and spring-loaded arm support (SLA). During the experiments, the electromyograms (EMG) of four muscles were simultaneously recorded: m. deltoideus anterior and lateralis, m. trapezius pars descendens and m. extensor carpi radialis brevis. Normalization was made against maximum isometric contraction. The mean values of the normalized EMG levels showed a reduced EMG level of the shoulder muscles when using arm supports in all the tasks, and for all muscles but the wrist extensor, compared to the EMG levels without arm supports. The horizontal movable support was more effective in reducing the EMG levels of the shoulder muscles than other arm supports, in tasks at table height. Thus, it is possible to reduce muscle activity of the shoulder region by using arm supports. Further research is needed to make biomechanical calculations to compare the EMG level of these muscles using suspension and the effects of inclination of work task. PMID- 9336108 TI - Prospective study of biofeedback for treatment of constipation. AB - PURPOSE: This study was designed to evaluate prospectively the results of pelvic floor physiotherapy with the aid of biofeedback in a heterogeneous group of patients with intractable constipation. METHODS: Biofeedback was used to treat 19 patients (age range, 16-78 (median, 63) years) with intractable constipation. Assessment, using visual linear analog scales of symptoms, was performed prospectively by an independent researcher. Biofeedback was performed by a physiotherapist, and patients were required to attend six sessions on an outpatient basis. The cause of constipation was heterogeneous, with no specific disorder being implicated on testing with anal manometry, defecating proctography, and colonic transit time. RESULTS: At six weeks, there was a median 27 percent (range, -8-93 percent) improvement in symptom scores. At six months, there was a median 23 percent (range, -54-64 percent) improvement in symptom scores. These were statistically significant compared with the scores at outset, six weeks (P = 0.0006), and six months (P = 0.012). However, only two (12.5 percent) patients at the six-month follow-up had an improvement of greater than 50 percent in their symptoms. CONCLUSION: Biofeedback is not recommended in the management of constipation. PMID- 9336109 TI - Results of biofeedback in constipated patients: a prospective study. AB - PURPOSE: The aims of this study were to assess the results of biofeedback treatment in constipated patients and to identify variables that might be used to predict the outcome. METHOD: Twenty-eight patients (5 men; median age, 46 (range, 22-72) years) with any degree of paradoxical activation measured with thin hook needle electromyography in the external sphincter or puborectalis muscle were included. The symptom duration varied between 1 and 30 (median, 9) years. The patients had eight outpatient training sessions with electromyography-based audiovisual feedback. All patients were followed up prospectively with a validated bowel function questionnaire from which a symptom index was created. RESULTS: At three months, nine patients had no improvement and underwent other treatments. The remaining 19 patients were followed up for a median of 14 (range, 12-34) months. Twelve patients (43 percent) stated they had improved rectal emptying. A good result was associated with increased stool frequency (P < 0.05), improved symptom index (P < 0.01), and reduction of laxative use (P < 0.05). A long symptom duration, a high pretreatment symptom index, and laxative use were related to a poor result (P < 0.01-0.05). The improved group had less perineal descent (P < 0.05), and a prominent puborectalis impression on defecography tended to be more common (P = 0.06). CONCLUSION: With the use of wide inclusion criteria, biofeedback was successful in 43 percent of patients, with a treatment effect lasting at least one year. The results suggest that biofeedback should be used as the initial treatment of constipated patients with a paradoxical puborectalis contraction. PMID- 9336111 TI - Character of the invasive margin in colorectal cancer: does it improve prognostic information of Dukes staging? AB - PURPOSE: The clinical significance and prognostic value of the histopathologic parameters used in both the Dukes and Jass classifications were evaluated to select those with an independent effect on survival after radical surgery for colorectal cancer. METHODS: The depth of local spread (limited to the bowel wall or extended beyond it), the number of metastatic lymph nodes (none, 1-4, more than 4), the character of the invasive margin (pushing or infiltrating), and the presence or absence of conspicuous peritumoral lymphocytic infiltration were assessed in 235 patients who had undergone radical resection for colorectal cancer. The influence of these variables on survival was studied by univariate and multivariate analysis. RESULTS: No significant difference in survival was found between patients with conspicuous peritumoral infiltrate and those without it; moreover, multivariate analysis failed to show any independent prognostic value for either lymphocytic infiltration or depth of local invasion. However, the character of the invasive margin and the number of metastatic lymph nodes were identified as the only variables with any independent importance on survival. Based on these data, a new prognostic model may be proposed; it uses the character of the infiltrative margin as a discriminating factor among patients within the lymph node-negative (Dukes A and B stages) and lymph node positive (Dukes C1 and C2 subsets) groups. A good prognosis for Dukes A, B, and C1 patients was associated with pushing tumors; C1 and C2 patients with infiltrating tumors had a poor prognosis. On the whole, the new prognostic model has allowed for the placement of 59.6 percent of our patients into groups that provide a confident prognosis. The clinical outcome of Dukes A and B patients with infiltrating tumors is still uncertain. CONCLUSIONS: The character of the invasive margin is an important prognostic factor in colorectal cancer. The association of this parameter with the traditional Dukes classification may provide additional useful prognostic information and aid in the selection of those patients who could most benefit from adjuvant therapy. PMID- 9336110 TI - Clinical and genomic influence of sulindac on rectal mucosa in familial adenomatous polyposis. AB - PURPOSE: A study was performed to evaluate the antiproliferative effects of low doses of the nonsteroidal drug, sulindac, on adenomas and rectal mucosa in familial adenomatous polyposis and to analyze the influence on tumor-suppressor genes and on apoptosis. METHODS: This was a prospective, controlled, nonrandomized Phase II dose-finding study for sulindac. The study group (n = 28) and control group (n = 10) underwent colectomy and ileorectal anastomoses, with repeated proctoscopy with endoluminal ultrasound and biopsies every three months. Dose-reduction of sulindac according to adenoma reversion was predetermined. Proliferation marker, Ki-67 (MIB1 and 5), on frozen or paraffin sections evaluated the antiproliferative effects; mutant p21ras, pantropic p53, mutant p53, and anti-bcl-2 were performed as enzyme-linked immunosorbent assay procedures and/or immunohistochemistry on paraffin sections. RESULTS: All patients responded to sulindac after 24 weeks (at the latest). There was a significant reduction of adenomas and dose reduction to 67 mg/day after three years of therapy (Mann's test for trend, P < 0.001). Results consisted of 78 percent complete reversions, 22 percent partial reversions of adenomas at latest re-examination, and no influence on upper gastrointestinal tract adenomas. No influence was detected on repeated hemograms, liver, or renal function at high or low doses. There was a permanent antiproliferative effect (Ki-67) of low-dose sulindac, significant blocking of ras mutation activation, and a significant difference of untreated and treated mucosa in mutant p53 content (Wilcoxon's or Kruskal-Wallis each, P < 0.05). Reverse correlation of anti-bcl-2 and p53 immunostaining on mucosa sections was an indication of adenoma relapse. CONCLUSIONS: Low-dose antiproliferative sulindac therapy is highly effective in adenoma reversion in familial adenomatous polyposis patients. Sulindac shows influence on tumor-suppressor genes and on apoptosis markers. An immunostaining correlation indicates adenoma relapse in flat microadenomas in advance of macroscopic appearance. Low-dose sulindac treatment may develop into an additive permnanent therapy for colectomized familial adenomatous polyposis patients. PMID- 9336112 TI - Use of brachytherapy in management of locally recurrent rectal cancer. AB - PURPOSE: Locally recurrent rectal cancer is associated with poor quality of life and has justified aggressive surgical and adjuvant approaches to control the disease. This study was designed to evaluate if the use of brachytherapy in association with wide surgical excision (debulking operation) can offer reasonable palliation for patients with locally recurrent rectal cancer. MATERIALS AND METHODS: Patients with biopsy-proven locally recurrent rectal cancer who were not candidates for intraoperative radiation therapy and who were previously considered as having unresectable tumors were included in the study and were followed-up from May 1981 to November 1990. All of them had undergone laparotomy and had either radical or debulking surgical resection performed. At the same time, brachytherapy was used with temporary or permanent implant of seeds of iridium-192 or iodine-125. RESULTS: Thirty patients were included. Patients ranged in age from 28 to 74 years, and 16 patients were female. No mortality was observed, and morbidity was low (small-bowel obstruction (1 patient), intestinal fistula (1 patient), and urinary fistula (1 patient). Histologic examination of the specimen showed gross residual disease in 67 percent of patients and microscopic disease in 25 percent of patients. Long-term follow-up was possible in 28 patients. Mean follow-up and local control were, respectively, 26.5 months and 37.5 percent for gross residual disease and 34 months and 66 percent for microscopic residual disease. Eighteen patients (64 percent) had locally recurrent rectal cancer under control at the time of the last follow-up, with seven patients (25 percent) having no evidence of local or distant recurrence. CONCLUSION: This is the first report of brachytherapy for locally recurrent rectal cancer. This appears to offer a therapeutic alternative to patients who are not candidates for intraoperative radiation therapy. Surgical morbidity and mortality are acceptable. Local control in 18 patients (64 percent) is comparable with intraoperative radiation therapy or more morbid surgical alternatives. Cancer-related deaths are most often related to disseminated disease, which suggests the need for systemic therapy in addition to brachytherapy. PMID- 9336113 TI - Influence of tumor position on accuracy of endorectal ultrasound staging. AB - Endorectal ultrasound is a well-established method of preoperative staging of rectal neoplastic lesions. PURPOSE: This study was undertaken to evaluate whether tumor site (in terms of height) and position (with respect to the rectal circumference) have an influence on the reliability of endoluminal ultrasound staging. METHODS: From January 1991 to May 1996, 154 consecutive patients with a total of 162 rectal tumors were examined preoperatively using endorectal ultrasound. Apart from staging all tumors using the uT/uN classification, tumor level and tumor position were recorded prospectively. Neoplasms were subdivided into low rectal (0-6 cm from the anal verge), mid rectal (7-12 cm), and higher lesions (> 12 cm). Furthermore, the lumen was divided into an anterior, left lateral, posterior, and right lateral position, and all tumors, apart from circular lesions (n = 9), were subclassified accordingly. RESULTS: Overall, we found 40 (25 percent) adenomas, 15 (9 percent) T1, 29 (18 percent) T2, 67 (41 percent) T3, and 11 (7 percent) T4 lesions. Overall accuracy was 78 percent. Staging accuracy for low rectal tumors (n = 41) was 68 percent, whereas 76 and 88 percent of mid (n = 96) and high (n = 25) neoplasms were staged correctly, respectively. The difference was not statistically significant. With regard to position, 47 tumors were situated anteriorly (77 percent accuracy), 42 in the left lateral position (69 percent accuracy), 33 posteriorly (73 percent accuracy), and 31 in the right lateral position (81 percent accuracy). Differences did not reach statistical significance. CONCLUSION: Endorectal ultrasound is currently the best method for preoperative assessment of the depth of infiltration of rectal tumors. However, rectal anatomy seems to affect staging accuracy in the lower rectum because the structure of the ampulla recti renders endosonographic examination more difficult. In addition, endosonographic layers are less well defined at this level. Both factors contribute to a lower reliability and predictive value of endorectal ultrasound staging in the lower rectum, although statistical significance was not reached in this study. On the other hand, tumor position with respect to rectal circumference does not influence the predictive value of endorectal ultrasound. PMID- 9336114 TI - Paget's disease of the perianal region--an aggressive disease? AB - BACKGROUND: Perianal Paget's disease is a rare entity, often associated with internal malignancies and a poor prognosis. METHODS: A chart review of patients with perianal Paget's disease who presented consecutively to Mayo during 25 years (starting in January 1970) was made. Patients included had Paget's disease located in or around the anus (3 cm). Patients were excluded for evidence of spread of vulvaperineal lesions or pagetoid extension of a rectal adenocarcinoma. Histology slides were reviewed, and immunohistochemistry was applied to confirm diagnoses. Follow-up was updated in all patients. Recurrence and survival curves were generated by the Kaplan-Meier method. Survival was compared with an age matched population by the log-rank test. RESULTS: Thirteen patients, eight females, were diagnosed (age +/- standard deviation of 68.3 +/- 10.6 years). All histologic diagnoses were confirmed with immunohistochemical staining results. Mean follow-up was 6.7 years, 8.8 for living patients. One patient had associated extramammary Paget's disease (scrotum). Lesions were located randomly at the dentate line, anal verge, and/or perianal area. Four patients had associated carcinomas; none of them were visceral. Eleven patients underwent local resection, without adjuvant therapy. Almost all recurrences were treated by wider local excision. The five-year recurrence rate was 61 percent. Overall five-year and ten-year survival was 67 percent, no different from the age-matched population (P = 0.546). CONCLUSIONS: These results do not reflect an aggressive nature of perianal Paget's disease, despite a high rate of local recurrence. Both primary lesions and recurrences are susceptible to treatment by wider local resection. Long-term survival is no different from that of the normal age-matched population. PMID- 9336115 TI - Fluorodeoxyglucose whole-body positron emission tomography in colorectal cancer patients studied in routine daily practice. AB - PURPOSE: To evaluate the routine clinical value of attenuation-corrected whole body fluorodeoxyglucose positron emission tomography in colorectal cancer, a total of 59 patients who were referred for evaluation of suspected or proven colorectal cancers were studied. METHODS: Positron emission tomography scans were recorded using a Siemens ECAT Exact 921/47. RESULTS: Median follow-up after the positron emission tomography study was 11 (mean, 12.3; range, 1-21) months. According to computed tomography, coloscopy, and ultrasound, we recorded eight apparently false-positive results. During later follow-up, however, three of those cases, which were negative with computed tomography, magnetic resonance imaging, sonography, or laparoscopy, turned out to be true-positive instead. In 3 patients, a primary colorectal cancer was suspected; in 26 patients, a recurrence of colorectal cancer was suspected. Eight patients were studied for follow-up after the history of colorectal cancer with no suspicion of recurrence. In 12 patients, the rise of serum tumor marker concentrations was the reason for the positron emission tomography study; 12 patients with known metastatic disease were also included ("restaging"). With regard to the entire patient population, we found an overall sensitivity of 100 percent, a specificity of 67 percent, and positive and negative predictive values of 92 and 100 percent, respectively. Being merely confirmative with respect to tumor recurrence or distant metastases in the majority of patients, positron emission tomography revealed a primary tumor in one patient and confirmed metastatic foci in several patients that had not been delineated by other imaging modalities. CONCLUSION: A whole-body positron emission tomography scan provides optimum conditions to locate metastatic lesions that might not be seen otherwise. There is a trend showing that positron emission tomography diagnostics as a consequence of early increased tumor markers is a highly sensitive combination, because computed tomography and magnetic resonance imaging were not as sensitive in early recurrences. Positron emission tomography, as performed in daily clinical practice, proved to be a powerful diagnostic tool in our subset of colorectal cancer patients. PMID- 9336117 TI - Pudendal neuropathy is the only parameter differentiating leakage from solid stool incontinence. AB - PURPOSE: Fecal incontinence may occur in several forms. Although some patients are grossly incontinent, other patients experience only leakage. In patients with gross incontinence, severity can range from the mildest forms (limited to loss of control of flatus) to the most severe forms (involving loss of solid stool). This study was undertaken to determine which physiologic parameters differentiate female patients with incontinence of solid stool from patients with control of formed stool and incontinence limited to seepage. METHODS: Thirty-eight consecutive female patients with a primary complaint of seepage or solid stool incontinence were evaluated using water perfusion manometry, balloon inflation assessment of rectal sensitivity, and pudendal nerve terminal motor latency. A prospectively maintained database was used for collection of data. The findings in the two patient groups were compared with patients in a group of normal control individuals. Ages of the women in the three groups were similar. RESULTS: Both groups of patients demonstrated statistically significant (P < 0.05) decreases in rest and squeeze sphincter lengths, pressures, and pressure volumes compared with normal volunteers. The patients also had significantly more asymmetric high-pressure zones and hypersensitive rectums. No significant difference between the two groups of incontinent patients could be identified using any of these parameters. Significant differences between the groups were found in pudendal nerve function. The distal rectoanal excitatory reflex was abnormal in 58.1 percent of grossly incontinent women compared with 28.6 percent of patients with leakage (P < 0.05). The majority of patients with leakage alone (65 percent) had normal pudendal nerve terminal motor latency, whereas only 22.6 percent of women with gross fecal incontinence had normal pudendal nerve terminal motor latency bilaterally (P = 0.01). CONCLUSIONS: Normal bilateral pudendal nerve function can partially compensate for abnormal sphincter symmetry and function, permitting women with grossly abnormal parameters to maintain control of bowel movements. It remains to be seen whether, with advancing age, patients with leakage will have development of slowed pudendal nerve conduction and, if so, whether their condition will progress to gross incontinence. PMID- 9336116 TI - Effect of high and intermediate ligation on survival and recurrence rates following curative resection of colorectal cancer. AB - PURPOSE: How wide excision of the regional mesenteric lymphatic drainage influences survival and recurrence rates following curative resection of colorectal cancers needs to be more clearly defined. METHODS: A series of 2,409 consecutive patients undergoing curative resections with detailed descriptions of the operative procedure and the lymphatic drainage in the surgical specimens provided a unique database to provide meaningful comparisons between high and intermediate level ligation. RESULTS: High ligation made a statistically significant difference in the death rate from recurrent cancer in patients with Dukes B, AC, and C1 cancers. Based on cancer-related deaths, the probability of five-year survival rate increased with high ligation from 73.9 to 84 percent in patients with Dukes B colon cancers and from 49.0 to 58.6 percent in patients with Dukes C1 colon cancers. In patients with Dukes AC cancers, high ligation increased the five-year survival rate from 64.9 to 80.4 percent. In patients with Dukes C cancers with involved middle level lymph nodes, the five-year survival rate increased from 20.5 to 33 percent and the death rate from recurrent cancer fell from 77 to 59 percent with high ligation. In patients with Dukes AC cancers with four or less involved nodes, the five-year survival rate was increased by high ligation from 50 to 78.6 percent in the colon and from 40 to 71.4 percent in the rectum. When more than four lymph nodes were involved, the survival rate was unaffected by the level of ligation. Although high ligation reduced distant recurrences, its greatest effect was observed in the incidence of local and suture line recurrence. The five-year local recurrence rate in patients with Dukes B who were managed by high ligation was 11.4 percent compared with 18.7 percent with intermediate ligation. In patients with Dukes C cancer, the local recurrence rate was 20.8 percent five years following high ligation compared with 30.7 percent for intermediate ligation. In patients with Dukes B cancer who were undergoing curative resections, the incidence of suture line recurrence was 3.9 percent following high ligation compared with 5.5 percent following intermediate ligation. In patients with Dukes C cancer, the incidence of suture line recurrence was 6.9 percent with high ligation and 11.4 percent with intermediate ligation. CONCLUSION: In certain stages of colorectal cancer, the more extensive resection of mesenteric lymphatic drainage associated with high ligation appears to increase the survival rate and reduce the recurrence rate following curative resections. PMID- 9336118 TI - Long-term results of suture rectopexy in patients with fecal incontinence associated with incomplete rectal prolapse. AB - Suture rectopexy is the recommended therapy for complete rectal prolapse that is associated with fecal incontinence. It has been suggested that correction of an incomplete rectal prolapse is also worthwhile for patients with fecal incontinence. PURPOSE: Aims of this study were 1) to evaluate the clinical outcome of suture rectopexy in a consecutive series of patients with incomplete rectal prolapse associated with fecal incontinence, and 2) to compare these results with those obtained from patients with complete rectal prolapse. METHODS: Between 1979 and 1994, suture rectopexy was performed in 13 incontinent patients (3 males; median age, 65 (range, 45-77) years) with incomplete rectal prolapse (Group I) and in 24 incontinent patients (21 females; median age, 71 (range, 24 86) years) with complete rectal prolapse (Group II). RESULTS: After a median follow-up of 67 months, continence was restored in 5 of 13 (38 percent) patients with incomplete rectal prolapse and in 16 of 24 (67 percent) patients with complete rectal prolapse. In both groups, all male patients became continent. CONCLUSIONS: For the majority of incontinent patients with incomplete rectal prolapse, a suture rectopexy is not beneficial. The clinical outcome of this procedure is only good in incontinent patients with complete rectal prolapse. Based on these data, it is questionable whether incomplete rectal prolapse plays a causative role in fecal incontinence. PMID- 9336119 TI - Colorectal function in patients with spinal cord lesions. AB - PURPOSE: This study was designed to describe the frequency and severity of colorectal problems among patients with spinal cord lesions and to determine whether these problems are associated with age, gender, time since the lesion, and level and severity of the lesion. PATIENTS AND METHODS: A detailed questionnaire describing colorectal and bladder function was sent to all 589 members of The Danish Paraplegic Association; 424 responded (72 percent). RESULTS: Only 19 percent felt a normal desire to defecate, whereas the remaining patients felt no desire to defecate (38 percent) or a combination of abdominal discomfort (37 percent) and headache, physical uneasiness, and perspiration (25 percent). Digital stimulation of the anal canal before defecation or digital evacuation of the rectum was used regularly by 65 percent of patients. Fecal incontinence was experienced by 75 percent of patients; however, most patients only had a few episodes of fecal incontinence each month (15 percent) or each year (56 percent). Overall, 39 percent of patients reported that colorectal dysfunction caused some or major restrictions on social activities or on their quality of life, and 30 percent regarded colorectal complaints to be worse than both bladder and sexual dysfunction. The severity of most symptoms was significantly correlated with the severity of the lesion, and the self-reported impact on social activities or quality of life was significantly more severe among women than men. CONCLUSION: Colorectal dysfunction is very common among spinal cord-injured patients, often causing restriction on social activities and quality of life. Therefore, these problems deserve more attention in the treatment of spinal cord-injured patients. PMID- 9336120 TI - Effect of fibrin glue on irradiated colonic anastomoses. AB - INTRODUCTION: The present study was planned to research the effects of fibrin glue on irradiated colonic anastomoses. METHOD: The effect of fibrin glue on irradiated colonic anastomoses was investigated in four identical groups of rats. In Group I (control group) colonic anastomoses were performed without radiotherapy; in Group II, colonic anastomoses were performed five days after radiotherapy; in Group III, fibrin glue was applied to anastomotic line without radiotherapy; in Group IV, fibrin glue was applied to anastomotic line with radiotherapy. The healing of left colonic anastomoses was evaluated through the bursting pressure of the anastomotic segment and the hydroxyproline contents of the anastomosis. RESULTS: Measurements done on the fourth postoperative day revealed that anastomotic healing was impaired in rats that underwent radiotherapy (P < 0.001); fibrin glue had no effect on anastomotic healing in groups with or without radiotherapy. CONCLUSION: In the early phases of anastomotic healing, fibrin glue cannot help remove unwanted effects of preoperative radiotherapy. PMID- 9336121 TI - Pathogenetic implications of DNA nondiploidy in colorectal cancers. AB - PURPOSE: Several studies propose that proximal and distal colorectal cancers have a different pathogenesis. We tested the hypothesis using flow cytometric DNA analysis. METHODS: DNA analysis was performed in 719 patients with colorectal cancer. In addition, histopathologic data were re-evaluated in a blinded fashion by a single pathologist. RESULTS: Distal tumors were more often nondiploid than were proximal tumors (61 vs. 49 percent; P = 0.015). Compared with the proximal tumor, distal tumors were smaller (P = 0.0001) and had less desmoplastic reaction (39 vs. 53 percent; P = 0.0001). Tumor location had no significant associations with the remaining parameters, including mucin production, perineural invasion, blood/lymphatic vessel invasion, lymphocytic infiltration, histologic grade, tumor stage, gross appearance, age, and gender. CONCLUSIONS: The unequal distribution of ploidy suggests distinct pathogenetic mechanisms at proximal and distal sites. PMID- 9336122 TI - Efficiency and productivity of a sheathed fiberoptic sigmoidoscope compared with a conventional sigmoidoscope. AB - PURPOSE: The aim of this study was to measure and compare time and productivity between a new sheathed flexible sigmoidoscope and a traditional fiberoptic flexible sigmoidoscope relative to labor and cost analysis. METHODS: Two flexible sigmoidoscopes, the Vision Sciences sigmoidoscope using a protective sheath covering requiring removal and replacement between procedures and a conventional flexible sigmoidoscope requiring meticulous cleaning using a washer and high level disinfection, were compared. Sigmoidoscope preparation was defined as the average time between the procedures (reprocessing, start to finish) and was measured by an independent nonmedical timekeeper JG). The parameter recorded was scope reprocessing time. RESULTS: Ten procedures were performed using the sheathed flexible sigmoidoscope system compared with nine using a conventional sigmoidoscope. Scope performance and endoscopic visualization for both systems were comparable. The average reprocessing time was 46.8 minutes for the conventional sigmoidoscope vs. 4.9 minutes for the sheathed sigmoidoscope (P < 0.0001). The average time saved was 9.5 times greater with the sheathed flexible sigmoidoscope system than with the conventional sigmoidoscope. CONCLUSION: The almost tenfold difference in the time saved using the sheathed flexible sigmoidoscope system represents increased productivity and potentially decreased overall labor cost. By reducing endoscope turnover time, this new sheathed system can reduce or even eliminate the need for backup endoscopes and endoscope washers and potentially allow better use of nursing staff. PMID- 9336123 TI - Incarcerated rectal prolapse--rupture and ileal evisceration after failed reduction: report of a case. AB - We report a case of incarcerated rectal prolapse that could not be reduced after using the previously described application of ordinary table sugar. Gentle pressure caused the prolapsed rectum to perforate, and the small bowel herniated through the tear. This is only the second case reported in the literature of an ileal herniation through a perforated rectum after an attempted reduction of an incarcerated prolapse. It is the only reported case occurring after sugar application and the 42nd case of ileal herniation through the rectum from all causes. PMID- 9336124 TI - Perforated rectal lymphoma in a renal transplant recipient: report of a case. AB - PURPOSE: We report the case of a renal transplant recipient with rectal lymphoma manifested by sudden onset of abdominal pain from a perforated rectum who was treated successfully with prompt surgical resection and reduction of immunosuppressants. METHODS: An emergent anterior resection with Hartmann's procedure was done. Immunosuppressants were drastically reduced by discontinuation of cyclosporine. RESULTS: Pathologic examination showed diffusely infiltrated large-cell malignant lymphoma with an immunoblastic feature. The patient has been followed-up for four years, with no tumor recurrence or graft rejection. CONCLUSION: Rectal lymphoma, although rare, should be kept in the list of differential diagnoses for transplant recipients who exhibit lower gastrointestinal bleeding, intestinal obstruction, or abdominal pain. PMID- 9336125 TI - Artificial anal sphincter. PMID- 9336126 TI - Laparoscopic sigmoid colectomy. PMID- 9336127 TI - Perineal resection for colorectoanal intussusception. PMID- 9336128 TI - Stage III nonsmall cell lung cancer: still a challenge for the radiation oncologist. PMID- 9336130 TI - Hyperfractionated accelerated radiation therapy for non-small cell lung cancer: clinical phase I/II trial. AB - PURPOSE: In an attempt to improve local control and survival of nonsmall cell lung cancer (NSCLC), hyperfractionated accelerated radiation therapy (HART) was carried out as a clinical phase I/II trial. METHODS AND MATERIALS: HART was delivered by 1.1 Gy/fraction, three fractions per day with intervals of 4 h and five treatment days per week. The clinical tumors were irradiated to 74.3 Gy (72.6-75.9)/66-69 fx, 33 days (29-40) (not corrected for lung density), and the subclinical lesions, to 50.0 Gy (48.4-50.6)/44-46 fx, 33 days (29-40). Sixty-nine patients with NSCLC were enrolled in this study. Nine patients were withdrawn from the study during HART due to different reasons. Sixty patients formed the study for outcome analyses. They were 57 males and 3 females with median age of 61 years (21-77). There were 41 cases of squamous cell carcinoma, 15 cases of adenocarcinoma, and 4 cases of large cell carcinoma. Overall, favorable patients (KPS > or = 70, weight loss < 5% and Stages I, II, IIIa) accounted for 73% (44 of 60) of all patients. Forty-four patients (73%) received adjuvant chemotherapy (DDP + VP16) with median cycles of 1.8 before and/or after HART. In order to compare the outcome of HART with conventional irradiation, 50 NSCLC patients treated by conventional fractionated irradiation (CFI) during the same period were chosen as the basis to evaluate relative effects of HART. They derived from the control group of another clinical trial of hyperfractionated irradiation for NSCLC in the same department. They received median tumor dose of 63.9 Gy (62.8 65.0)/34 fx (32-36), 48 days (45-53). RESULTS: 1. Acute and late complications: (a) In HART, 87% of patients (52 cases) developed acute radiation esophagitis: Grade 1-2, 46 cases (77%) and Grade 3, 6 cases (10%), at 2.5 weeks (2-3.5 weeks) after HART began. Five patients with Grade 3 esophagitis had their HART interrupted for <7 days. In CFI, esophagitis was much less (44%,p < 0.05) with 38% of Grade 1-2 and 6% of Grade 3. (b) In HART, acute pulmonary symptoms (RTOG Grade 1-2) occurred in 17% (10 cases), and acute radiation pneumonitis (Grade 3), in 8% (5 cases), while in CFI, they were 24% and 2% (p > 0.05), respectively. Late lung fibrosis (RTOG Grade 1-2) appeared in 20% (12 cases), whereas 18% in CFI (p > 0.05). (c), No other severe acute or late complications have been observed so far in HART. 2. Immediate response. In HART, 20% of patients (12 cases) achieved CR, 60% (36 cases), PR and 20% (12 cases), NR or PD. In CFI, the above three percentages were 10, 28, and 62%, respectively (p < 0.001). 3. Follow up. The 1-, 2-, and 3-year actuarial survivals were 72, 47, and 28% for HART, and 60, 18, and 6% for CFI, respectively (p < 0.001). Better local control was seen in HART than in CFI with 1-, 2-, and 3-year local control rates being 71, 44, 29%, and 60, 20, and 5%, respectively (p = 0.001). Distant metastases developed less in HART than in CFI. The 1-, 2-, and 3-year distant metastasis rates were 23, 36, and 50% in HART, but 30, 48, and 80% in CFI (p = 0.021). CONCLUSION: 1. HART could be tolerated by most of the favorable NSCLC patients. The predominant complication was acute esophagitis. No other severe acute or late complications have been observed so far. 2. HART resulted in better survivals and local controls, and less distant metastases than CFI. PMID- 9336129 TI - Induction cisplatin/vinblastine and irradiation vs. irradiation in unresectable squamous cell lung cancer: failure patterns by cell type in RTOG 88-08/ECOG 4588. Radiation Therapy Oncology Group. Eastern Cooperative Oncology Group. AB - PURPOSE: To analyze disease failure patterns by pretreatment characteristics and treatment groups in a prospective randomized trial. METHODS AND MATERIALS: Patients with medically inoperable Stage II, unresectable IIIA and IIIB nonsmall cell lung cancer with KPS > or =70 and weight loss < or =5% were randomized to one of three treatment groups: standard radiation therapy with 60 Gy at 2.0 Gy per day (STD RT), induction chemotherapy with cisplatin 100 mg/m2 days 1 and 29 with vinblastine 5 mg/m2 weekly for 5 weeks followed by 60 Gy at 2.0 Gy per day (CT + RT), or hyperfractionated radiation therapy with 69.6 Gy at 1.2 Gy b.i.d. (HFX RT). Of 490 patients enrolled, 458 were evaluable. Minimum and median periods of observation for this analysis were 4 years and 6 years, respectively. RESULTS: Pretreatment characteristics were equally distributed. Toxicities were previously reported. Median survival rates were 11.4, 13.6, and 12.3 months for STD RT, CT + RT, and HFX RT, respectively (log rank p = 0.05, Wilcoxon p = 0.04). Survivals were 20, 31, and 24% at 2 years, and 4, 11, and 9% at 4 years in the STD RT, CT + RT, and HFX RT groups, respectively. There were no differences in local tumor control rates among the treatments. Patterns of first failure showed less distant metastasis (DM) (other than brain) for CT + RT compared to the RT alone arms (p = 0.04). Within squamous cell carcinoma (SCC), DM (other than brain) rates were 43%, 16%, and 38% in SCC for STD RT, CT + RT, and HFX RT, respectively (p = 0.0015). Patients with peripheral/chest wall lesions were significantly more likely to fail first in the thorax when treated on STD RT compared to CT + RT and HFX RT (p = 0.009). Survival rates were similar among the treatment arms for patients with squamous cell carcinoma. Among patients with nonsquamous cell carcinoma, failure patterns did not differ by treatment group, but survival was significantly better in those who were treated by induction chemotherapy (p = 0.04). CONCLUSION: Patients with squamous cell carcinoma treated on the CT + RT arm had a significant reduction of first DM other than brain, but there was difference in survival. Survival favored CT + RT in nonsquamous carcinoma despite similar failure patterns. Reasons for improved survival with CT + RT in NSCLC are not yet available. PMID- 9336131 TI - Is the time interval between surgery and radiotherapy important in operable nonsmall cell lung cancer? A retrospective analysis of 340 cases. AB - PURPOSE: To evaluate the influence of prognostic factors in postoperative radiotherapy of NSCLC with special emphasis on the time interval between surgery and start of radiotherapy. METHODS AND MATERIALS: Between January 1976 and December 1993, 340 cases were treated and retrospectively analyzed meeting the following criteria: complete follow-up; complete staging information including pathological confirmation of resection status; maximum interval between surgery (SX) and radiotherapy (RT) of 12 weeks (median 36 days, range 18 to 84 days); minimum dose of 50 Gy (R0), and maximum dose of 70 Gy (R2). Two hundred thirty patients (68%) had N2 disease; 228 patients were completely resected (R0). One hundred six (31%) had adenocarcinoma, 172 (51%) squamous cell carcinoma. RESULTS: In univariate analysis, Karnofsky performance status (90+ >60-80%; p = 0.019 log rank), resection status stratified for nodal disease (R+ 30 Gy (p < 0.05). CONCLUSION: The extent of alteration in whole-lung function (symptoms or PFT changes) appears to be related to both dose-volume and pre-RT PFT parameters. The data suggest that no one variable is likely to be an adequate predictor and that multivariate predictive models will be needed. Additional studies are underway to develop better predictive models that consider physical factors such as the DVH and regional perfusion, as well as biological/clinical factors such as pre-RT PFTs and TGF-beta1. PMID- 9336134 TI - A randomized phase III study of accelerated hyperfractionation versus standard in patients with unresected brain metastases: a report of the Radiation Therapy Oncology Group (RTOG) 9104. AB - PURPOSE: To compare 1-year survival and acute toxicity rates between an accelerated hyperfractionated (AH) radiotherapy (1.6 Gy b.i.d.) to a total dose of 54.4 Gy vs. an accelerated fractionation (AF) of 30 Gy in 10 daily fractions in patients with unresected brain metastasis. METHODS AND MATERIALS: The Radiation Therapy Oncology Group (RTOG) accrued 445 patients to a Phase III comparison of accelerated hyperfractionation vs. standard fractionation from 1991 through 1995. All patients had histologic proof of malignancy at the primary site. Brain metastasis were measurable by CT or MRI scan and all patients had a Karnofsky performance score (KPS) of at least 70 and a neurologic function classification of 1 or 2. For AH, 32 Gy in 20 fractions over 10 treatment days (1.6 Gy twice daily) was delivered to the whole brain. A boost of 22.4 Gy in 14 fractions was delivered to each lesion with a 2-cm margin. RESULTS: The average age in both groups was 60 years; nearly two-thirds of all patients had lung primaries. Of the 429 eligible and analyzable patients, the median survival time was 4.5 months in both arms. The 1-year survival rate was 19% in the AF arm vs. 16% in the AH arm. No difference in median or 1-year survival was observed among patients with solitary metastasis between treatment arms. Recursive partitioning analysis (RPA) classes have previously been identified and patients with a KPS of 70 or more, a controlled primary tumor, less than 65 years of age, and brain metastases only (RPA class I), had a 1-year survival of 35% in the AF arm vs. 25% in the AH arm (p = 0.95). In a multivariate model, only age, KPS, extent of metastatic disease (intracranial metastases only vs. intra- and extracranial metastases), and status of primary (controlled vs. uncontrolled) were statistically significant (at p < 0.05). Treatment assignment was not statistically significant. Overall Grade III or IV toxicity was equivalent in both arms, and one fatal toxicity at 44 days secondary to cerebral edema was seen in the AH arm. CONCLUSION: Although a previous RTOG Phase I/II report had suggested a potential benefit in patients with limited metastatic disease, a good Karnofsky performance status, or neurologic function when treated with an AH regimen, this randomized comparison could not demonstrate any improvement in survival when compared to a conventional regimen of 30 Gy in 10 fractions. Therefore, this accelerated hyperfractionated regimen to 54.4 Gy cannot be recommended for patients with intracranial metastatic disease. PMID- 9336135 TI - Cost-utility analysis of a malignant glioma protocol. AB - PURPOSE: To perform a cost-utility analysis utilizing a cooperative group protocol and constrasting the results with the published quality adjusted survival. METHODS AND MATERIALS: A cost-utility analysis was performed on Radiation Therapy Oncology Group (RTOG) protocol 83-02. The quality-adjusted survival has been published previously. Pretreatment tests and chemotherapy costs are not considered, as these were similar across all treatment arms. Payor costs are calculated from Federal Register data for Medicare Region IV. Global charges are used to calculate the professional and technical charges. Costs are measured in relative value units (RVUs) and are tabulated assuming equal treatment complexity for all treatment arms. RESULTS: The number of RVUs calculated for each arm were 48 Gy--166.65; 54.4 Gy--182.17; 64.8 Gy--232.53; 72.0 Gy--272.19; 76.8 Gy--287.11; and 81.6 Gy--302.63. The RVU/QALY for the <50-year-old group were 48 Gy--119.03; 54.4 Gy--100.65; 64.8 Gy--104.78; 72.0 Gy--90.73; 76.8 Gy- 193.99; and 81.6 Gy--165.37. The RVU/QALY for the >50-year-old group were 48 Gy- 198.39; 54.4 Gy--276.85; 64.8 Gy--426.57; 72.0 Gy--423.71; 76.8 Gy--703.70; and 81.6 Gy--519.10. Sensitivity analysis of one treatment plan, simulation, and set of blocks for the 48 Gy and 54.4 Gy arms decreased the RVU/QALY to 105.34 and 90.05, respectively. DISCUSSION: Our analyses shows the experimental arm with the lowest RVU/QALY is also the arm with the longest quality-adjusted survival. This arm had the most efficient use of resources in this protocol. Prospective collection of all pertinent cost data is required for comparison of one treatment against another. All cooperative group protocols should have prospective quality of life and economic endpoints to allow for comparison of the most cost efficient treatment method. PMID- 9336136 TI - Childhood optic chiasm gliomas: radiographic response following radiotherapy and long-term clinical outcome. AB - PURPOSE: In children with chiasmal gliomas, radiation therapy can arrest progressive visual and neurologic impairment. We examined the radiographic response and clinical outcomes after irradiation. METHODS AND MATERIALS: Forty two children (median age at diagnosis, 6.6 years) with chiasmal gliomas were managed as follows: 11 asymptomatic patients with neurofibromatosis-1 (NF-1) were observed only; 2 patients, less than 3 years old, underwent surgery and chemotherapy to delay irradiation; and 29 patients with progressive disease received radiation with or without prior surgery or chemotherapy. Time to radiographic response, long-term tumor control and late sequelae were reviewed for the 29 irradiated patients. RESULTS: The probability of at least 50% radiographic response at 24 months after irradiation was 18.1% and increased to 38.2% by 48 months and 45.9% by 60 months. By actuarial analysis, the median time for such radiographic response was 62 months. For the 29 irradiated patients, the 10-year freedom from progression and overall survival rates were 100% and 89%, respectively (median follow-up for surviving patients, 108 months). Stabilization or improvement in vision occurred in 81% of 26 evaluable irradiated patients. CONCLUSIONS: Notable radiographic response may be observed years after irradiation. Radiation therapy provides excellent long-term tumor control and vision preservation or improvement in the majority of patients with progressive chiasmal gliomas. PMID- 9336137 TI - Optimized lens-sparing treatment of retinoblastoma with electron beams. AB - PURPOSE: The ideal lens-sparing radiotherapy technique for retinoblastoma calls for 100% dose to the entire retina including the ora serrata and zero dose to the lens. Published techniques, most of which use photons, have not accomplished this ideal treatment. We describe here a technique that approaches this ideal configuration using electron beam therapy. METHODS AND MATERIALS: Dose-modeling calculations were made using a computer program built around a proprietary algorithm. This program calculates 3D dose distribution for electrons and photons and uses the Cimmino feasibility method for the inverse problem of beam weighting to achieve the prescribed dose. The algorithm has been verified in the ocular region by measurements in a RANDO phantom. To search for an ideal lens-sparing beam setup, a stylized phantom of an 8-month-old infant was generated with built in inhomogeneities, and a phantom of a 5-year-old child was generated from a patient CT series. RESULTS: Of more than 100 different beam setups tested, two 9 MeV electron beams at gantry angles plus and minus 26 degrees from the optic nerve axis achieved the best distribution. Both fields have a lens block and an isocenter between the globe and origin of the optic nerve. When equal doses are given to both fields, the entire extent of the retina (including ora serrata) received 100%, while the lens received 10% or less. CONCLUSION: The two-oblique electron-beam technique here described appears to meet most of the stringent dosimetry needed to treat retinoblastoma. It is suitable for a range of ages, from infancy to early childhood years. PMID- 9336138 TI - Do not miss a second (and possibly last) chance to cure Hodgkin's disease. PMID- 9336139 TI - Salvage radiotherapy for Hodgkin's disease following chemotherapy failure. AB - PURPOSE: This study aims to: 1) assess failure-free survival (FFS), overall survival (OS), and failure pattern after salvage radiotherapy (SRT) for patients with Hodgkin's disease (HD) who fail chemotherapy (CT); 2) identify patients suitable for SRT as an alternative to more aggressive salvage regimens. METHODS AND MATERIALS: Between 1978 and 1992, 52 patients with relapsed/refractory HD following 26 CT received SRT at the Peter MacCallum Cancer Institute. Patient characteristics at diagnosis were: median age (range 12-63); male-31, female-21; Stage I-4, II-16, III-25, or IV-7. Prior to SRT 27 patients had received the equivalent of both MOPP and ABV(D). The duration of initial complete response (CR) from CT was greater than 12 months in 22 patients. SRT (dose 34-42 Gy) was given to active disease sites. RESULTS: Five-year FFS and OS rates following SRT were 26 and 57%, respectively. Five-year FFS and OS rates of 36 and 75%, respectively, were achieved in patients who relapsed in supradiaphragmatic nodal sites without B symptoms; in a subset of patients with initial Stage I-II disease the FFS and OS rates were 50 and 86%, respectively. On multivariate analysis significant factors for FFS were B symptoms at the time of SRT (p = 0.003), extranodal involvement (p = 0.011) and histology (p = 0.018). For OS significant factors were B symptoms (p = 0.0007), age (p = 0.014) and number of prior CT regimens (p = 0.03). CONCLUSION: The relatively poor results of SRT in terms of FFS justify the use of alternative salvage strategies for most patients with Hodgkin's disease who fail CT. However, SRT offers a low morbidity, potentially curative option for a subset of patients. Our data suggest that patients most suitable for SRT are those with relapse in supradiaphragmatic nodal sites and no B symptoms. PMID- 9336140 TI - Long-term outcome of treatment for Ann Arbor stage 1 Hodgkin's disease: patterns of failure, late toxicity and second malignancies. AB - PURPOSE: Radiation therapy results in excellent short-term survival in patients with early-stage Hodgkin's disease. However, the optimal therapeutic scheme that achieves the highest disease-free survival with the minimum long-term toxicity is yet to be determined. An analysis of the patterns of failure and late complications after radiation therapy was conducted to address this question. METHODS AND MATERIALS: A retrospective study was conducted of 145 patients with Stage I Hodgkin's disease treated at M. D. Anderson Cancer Center from 1967 through 1987. Follow-up extended from a minimum of 30 to 339 months, with a median period of observation of 16.5 years. All the patients were treated with radiation therapy and, and 16 received combination MOPP-based chemotherapy as part of their initial treatment. The radiotherapy technique, was involved/regional in 71 (49%), extended in 62 (43%), and subtotal nodal irradiation in 12 patients. The median total dose was 40 Gy. RESULTS: The actuarial freedom from progression at 10 and 20 years was 76% and 69%, respectively. Forty of 145 patients relapsed (27.6%). The site of primary disease was cervical adenopathy in 30 (75%), axillary in 7 (17.5%), mediastinal in 2 patients and subdiaphragmatic in one patient. Twenty-two patients were treated with involved/regional technique (55%), 17 with extended (42.5%), and 1 with subtotal nodal irradiation technique. There were three in field and four marginal recurrences. Six relapses occurred in non-irradiated nodal regions at the same side of the diaphragm and 17 in non-irradiated transdiaphragmatic lymph nodes (57.5%). Nine patients (22.5%) relapsed with visceral disease. Nineteen patients (47.5%) relapsed within the first 2 years, 15 (37.5%) 3 to 10 years after diagnosis and the remaining 6 (15%) after 10 years. Eleven of 40 patients died of disease after the first or subsequent relapses (27.5%). Three of six patients with late relapses had progression in viscera but only two died with disease. Thirty-eight of 145 patients developed late toxicity from the treatment (26.2%). Twenty-three patients experienced ischemic heart disease (15.9%), only 13 of whom received mediastinal irradiation (9%). Fifteen patients developed secondary malignant solid tumors (10.3%). Nine of those (6.2%) occurred within the irradiation field (two were also treated with chemotherapy). Two additional patients, one of whom received chemotherapy as part of the initial treatment, died of acute myelogenous leukemia. Non-Hodgkin's lymphoma and lung cancer were the most common second malignancies. CONCLUSIONS: Limited field radiotherapy results in a significant number of relapses in non-irradiated, especially transdiaphragmatic lymph nodes. Subtotal nodal irradiation can prevent some relapses and therefore improve freedom from progression. Careful design of the treatment fields may decrease the risk of morbidity and mortality from coronary artery disease and second malignancies in early-stage Hodgkin's disease. Careful long-term surveillance may permit early detection and management of late relapses and treatment complications. PMID- 9336141 TI - High dose chemotherapy and stem cell rescue for aggressive non-Hodgkin's lymphoma: pattern of failure and implications for involved-field radiotherapy. AB - PURPOSE: To evaluate the pattern of failure and outcome of patients with aggressive non-Hodgkin's lymphoma (NHL) undergoing high-dose chemotherapy (HDCT) and autologous stem cell rescue (SCR) with an emphasis on the role of adjuvant involved-field radiotherapy (IFRT). METHOD AND MATERIALS: Fifty-three adult patients with aggressive NHL (46 intermediate-and 7 high-grade) underwent HDCT with SCR. All patients underwent induction chemotherapy prior to high dose intensification. Seven (13.2%) received IFRT to 10 disease sites either prior to or following HDCT. Indication included symptomatic or bulky disease, persistent disease, or to consolidate a complete response (CR). Sites of relapse were designated as old (involved prior to HDCT) or new (previously uninvolved). Median followup was 20.1 months (range, 1.2-69.3 months). RESULTS: The 4-year actuarial progression-free (PFS) and cause-specific (CSS) survivals of the entire group were 30.0 and 50.2%, respectively. Excluding toxic deaths, 24 patients (52.2%) relapsed. Sixteen (34.7%) failed in old and 15 (32.6%) in new sites. Patients treated with IFRT had a lower rate of relapse in old sites (0 vs. 41%) (p = 0.04) than patients treated with HDCT alone. Of the 141 sites present prior to induction, 127 (90.1%) were amenable to IFRT. Excluding irradiated sites, the overall 4-year local control (LC) of all amenable sites was 61.1%. Amenable sites failing to achieve a CR to induction had a poorer LC (32.0 vs. 95.1%) (p < 0.0001) than sites in CR. The 4-year LC of sites failing to achieve a CR to HDCT was 29.4%. Adjuvant IFRT improved the 4-year LC of all sites (100 vs. 61.1%) (p = 0.05), persistent sites following induction (100 vs. 32.0%) (p = 0.01) and persistent sites following HDCT (100 vs. 29.4%) (p = 0.01). Adjuvant IFRT was not associated with any untoward acute or late toxicity. CONCLUSIONS: The predominant site of relapse in patients with aggressive NHL undergoing HDCT and SCR is in sites of disease present prior to HDCT. However, the risk of relapse of prior disease sites varies greatly depending upon their response to chemotherapy. Sites at greatest risk are those failing to achieve a CR to induction regardless of their response to HDCT. IFRT is capable of reducing the high risk of relapse in these sites, the majority of which are amenable to IFRT. These results demonstrate a rationale for and possible benefit to IFRT in patients with aggressive NHL undergoing HDCT and SCR. PMID- 9336142 TI - Ductal carcinoma in situ detected in the mammographic era: an analysis of clinical, pathologic, and treatment-related factors affecting outcome with breast conserving therapy. AB - PURPOSE: We reviewed our institution's experience treating predominantly mammographically detected ductal carcinoma in situ (DCIS) with breast-conserving therapy (BCT) to determine if any clinical, pathologic, or treatment-related factors affected outcome. METHODS AND MATERIALS: From January 2, 1980 to January 6, 1992, 107 breasts in 105 patients were treated with BCT at William Beaumont Hospital, Royal Oak, MI. All patients underwent at least an excisional biopsy and 70 patients (65%) were reexcised. All patients received whole-breast irradiation to a median dose of 50.4 Gy (range 43.1 to 56.0 Gy). Ninety-nine patients (93%) received a supplemental boost to the tumor bed for a median total dose of 60.4 Gy (range 59.1 to 71.8 Gy) using either photons (2 patients), electrons (69 patients), or an interstitial implant (28 patients). RESULTS: With a median follow-up of 78 months, 10 patients have failed in the treated breast for a 5- and 10-year actuarial local control rate of 91.2 and 89.8%, respectively. Thirteen percent of the population have been followed for 10 years or more. Three recurrences were pure DCIS, and seven were invasive. All patients were salvaged with mastectomy. Nine patients remain without evidence of disease a median of 30.6 months after surgery. One patient failed distantly 36 months after local recurrence for an ultimate cause specific survival of 99%. Potential clinical (age, mammographic findings, method of detection, etc.), pathologic (nuclear grade, margins, etc.), and treatment-related factors (dose, boost technique, reexcision status, etc.) affecting outcome were analyzed. No variable was found to be associated with an ipsilateral breast tumor recurrence. However, when only recurrences that occurred within or immediately adjacent to the lumpectomy cavity were analyzed, both margin status and the extent of cancerization of lobules (COL) near the surgical margin were associated with the development of a local recurrence. CONCLUSIONS: Patients treated with BCT for predominantly mammographically detected DCIS achieve excellent rates of local control and overall survival. Both margin status and the extent of COL near the surgical margin appear to be associated with recurrences within or immediately adjacent to the lumpectomy cavity. These data suggest that careful attention to the completeness of surgical resection of DCIS is an important determinant of outcome. PMID- 9336143 TI - Preservation of cosmesis with low complication risk after conservative surgery and radiotherapy for ductal carcinoma in situ of the breast. AB - PURPOSE: Although the clinical outcome after treatment of ductal carcinoma in situ (DCIS) using breast-conservation surgery and radiation therapy has been well documented, little data has been reported on cosmetic outcome or treatment complications. Therefore, the present study was performed to evaluate cosmesis and complications after breast-conservation treatment for DCIS and to analyze various factors that might affect cosmesis and predispose to complications. METHODS AND MATERIALS: The records of 90 patients who were alive without evidence of disease with a 3-year minimum follow-up were evaluated for cosmetic results and treatment complications following breast-conservation surgery and radiation therapy for DCIS. Complete gross excision of the primary tumor had been performed in all patients. Additionally, 24 patients had undergone an axillary lymph node dissection in the earlier years of the study. The majority of the patients had received 50-50.4 Gy to the whole breast followed by an electron boost for a total dose of 60-66 Gy. RESULTS: The cosmetic results of 90 evaluable patients at 3 years were: excellent in 69 (77%), good in 19 (21%), and fair in 2 (2%). The cosmetic results of 64 evaluable patients at 5 years were: excellent in 46 (72%), good in 16 (25%), and fair in 2 (3%). Factors associated with worse cosmetic results were an increased volume of tissue excised (>70 cm3) and a negative ipsilateral breast biopsy after radiotherapy. Complications in the 24 patients with an axillary dissection were: arm edema (n = 6), cellulitis of the arm (n = 5), and axillary vein thrombosis (n = 1). Complications in the 66 patients without an axillary dissection were: cellulitis of the arm (n = 1) and cellulitis of the breast (n = 1). DISCUSSION: Breast-conservation surgery followed by radiation therapy achieved excellent or good cosmetic results in 98 and 97% of patients at 3 years and 5 years, respectively. Complications were associated primarily with axillary dissection, which is no longer standard practice, and complications were uncommon in patients without an axillary dissection. Therefore, patients currently treated for DCIS of the breast would be expected to have a high rate of excellent or good cosmetic outcome with a low risk of complications. PMID- 9336144 TI - Curative surgical resection following reirradiation for recurrent rectal cancer. AB - PURPOSE: In spite of adjunctive radiation and chemotherapy, 10 to 25% of patients with resected rectal cancer develop local recurrence in the pelvis. This study evaluates the potential for curative surgical resection of residual disease following reirradiation for recurrent rectal cancer. METHODS AND MATERIALS: Thirty-nine patients with recurrent adenocarcinoma of the rectum following prior adjunctive therapy underwent reirradiation of the pelvis with concurrent intravenous infusion of 5-fluorouracil. Median time to recurrence following initial treatment was 18 months. Prior radiation doses to the pelvis ranged from 40 to 66 Gy with a median of 50.4 Gy. Reirradiation doses ranged from 20 Gy to 49.2 Gy with a median total dose of 36 Gy. Eight to 12 weeks following reirradiation patients underwent surgical resection of disease. Thirty-one patients had gross total resection of tumor. RESULTS: Patients have been followed for 24 months to 75 months after reirradiation for recurrent rectal cancer with a median follow-up of 3 years. Reirradiation was well tolerated, with seven patients requiring a significant treatment break. Early termination of reirradiation occurred in five patients because of diarrhea, moist desquamation, or mucositis. No surgical mortality was observed. Postoperatively, two patients developed delayed wound healing. Late complications included six patients who developed small bowel obstruction with three patients developing a bowel fistula. The median survival of patients is 45 months, with a 5-year actuarial survival of 24%. Actuarial local control at 5 years was 45%. The rate of distant metastases was 17%. CONCLUSION: Selected patients with rectal cancer who develop recurrent disease following previous adjuvant therapy can undergo successful curative surgical resection following reirradiation/chemotherapy with significant long term survival. PMID- 9336145 TI - Time-dose considerations in the treatment of anal cancer. AB - PURPOSE: To analyze the impact of patient and treatment parameters in concurrent chemoradiation treatment for anal carcinoma. METHODS AND MATERIALS: Retrospective review of 50 MO anal cancer patients treated from 1984-1994. Most patients received concurrent 5-FU, mitomycin, and radiation. Local control and disease free/overall survival were determined and analyzed according to patient and treatment parameters. RESULTS: With 43 month median follow-up, projected overall survival is 66% at 5 and 8 years. Disease-free survival is 67% at 5 years and 59% at 8 years. Local control is 70% at 5 and 8 years. Doses of > or =54 Gy are associated with improved 5-year survival (84 vs. 47%, p = 0.02), disease-free survival (74 v. 56%, p = 0.09), and local control (77 vs. 61%, p = 0.04). Although local control, disease-free survival, and overall survival were improved in patients whose overall treatment time was <40 days, this was not statistically significant. Outcome in the four patients with pretreatment hemoglobin (Hgb) <10 appeared worse with 3-year overall survival 50 vs. 68% (p = 0.07), disease-free survival 0 vs. 67% (p = 0.11), and local control 0 vs. 74% (p = 0.05). Projected 5-year overall survival, relapse-free survival, and local control in 4 HIV(+) patients is 0, 75, and 75%. Multivariate analysis reveals that dose (p = 0.02) and Hgb (p = 0.05) independently affect local control, dose (p = 0.02) affects disease-free survival, and dose (p = 0.01), Hgb (p = 0.03), T-stage (p = 0.03), and HIV-status (0.07) independently influence overall survival. CONCLUSION: Radiation doses of > or =54 Gy are associated with significantly improved survival and local control in anal cancer patients treated with chemoradiation. Overall treatment times of less than 40 days are associated with a trend towards improved outcome, but this is not significant. Pretreatment hemoglobin <10 is associated with worse treatment outcome. Survival of HIV (+) patient is poor, but the majority of such patients in this series died of intercurrent disease with their anal carcinomas controlled by chemoradiation. PMID- 9336146 TI - The role of adjuvant radiotherapy in the treatment of resectable desmoid tumors. AB - PURPOSE: Desmoid tumors have a high propensity for local recurrence with surgical resection. There are many reports describing good responses of desmoid tumors to irradiation, but none have clearly established the indications for adjuvant radiotherapy in treating resectable desmoid tumors. METHODS AND MATERIALS: A retrospective analysis was performed on 61 patients with resectable desmoid tumor(s) who were treated at our institution from 1965 to February of 1992. Five patients had multifocal disease and are analyzed separately. Fifty-six patients had unifocal disease, of which 34 had positive surgical margins. Forty-five of the 56 patients with unifocal disease were treated with surgery alone, while 11 were treated with surgery plus adjuvant radiotherapy. Median follow-up was 6 years. Local control was measured from the last day of treatment, and all cases were reviewed by our Department of Pathology. RESULTS: Multivariate analysis of the 56 patients with unifocal disease revealed that positive margins independently predicted for local recurrence (p < or = 0.01). Only 3 of 22 patients with clear margins experienced a local recurrence, with a 6-year actuarial local control of 85%. Multivariate analysis of the 34 patients with positive margins revealed that adjuvant radiotherapy independently predicted for improved local control (p = 0.01), and patients with recurrent disease had a slightly higher risk of local recurrence (p = 0.08). The 6-year actuarial local control determined by Kaplan-Meier for patients with unifocal disease and positive margins was 32% (+/-12%) with surgery alone, and 78% (+/-14%) with surgery plus adjuvant radiotherapy (p = 0.02). Subgroup analysis of the patients with positive margins and recurrent disease revealed that those treated with surgery alone had a 6-year actuarial local control of 0% vs. 80% for those treated with surgery plus radiotherapy (p < or = 0.01). Patients with positive margins and primary disease had a trend towards improved local control with adjuvant radiotherapy, but this was not statistically significant. None of the patients treated with radiotherapy developed serious complications or a secondary malignancy. CONCLUSIONS: Margin status is the most important predictor of local recurrence for patients with resectable, unifocal desmoid tumor. Adjuvant radiotherapy is indicated in the treatment of patients with positive margins following wide excision of recurrent disease. The role of adjuvant radiotherapy in patients with positive margins following resection of primary disease is controversial, and should be based on a balanced discussion of the potential morbidity from radiotherapy compared to the potential morbidity of another local recurrence. Adjuvant radiotherapy is less likely to benefit those with clear margins due to the excellent results for these patients treated with surgery alone. The local control of desmoid tumor in the adjuvant setting is excellent with total doses ranging from 50-60 Gy, with acceptable morbidity. Field sizes should be generous to prevent marginal recurrences, and large volume MRIs of patients with extremity lesions should be used to identify those patients with multifocal disease. PMID- 9336147 TI - Pulmonary embolization of permanently implanted radioactive palladium-103 seeds for carcinoma of the prostate. AB - PURPOSE: It has been reported that permanently implanted iodine-125 seeds can embolize to the lungs. There is little data on the embolization of palladium-103 seeds. The purpose of this study is to collect and evaluate data on the embolization of Pd-103 seeds. METHODS AND MATERIALS: The records of 112 patients implanted with Pd-103 for carcinoma of the prostate were reviewed to systemically study the incidence and dynamics of pulmonary embolism of Pd-103 seeds. Five patients had no postoperative chest radiograph and were thus excluded, leaving 107 patients for review. RESULTS: Chest radiographs of 19 of the 107 patients showed embolized seeds in the lungs (18%). Two patients had three seeds each, nine patients had two seeds each; and in the remaining eight patients, a single seed migrated to the lungs. The seeds migrated mainly (84%) to the lower lobes. None of the eight patients who had their first postoperative chest radiograph on the day of the implant showed any embolized seeds. The embolized seed appeared only on subsequent chest radiographs taken 27 to 40 days later. Ten of the other 11 patients who had their first radiograph 1 to 97 days after brachytherapy had embolized seeds on their first chest radiograph. In the other patient, the embolized seed appeared only on a subsequent chest radiograph taken after 127 days. There were no clinical pulmonary or cardiac effects evident on routine follow-up of these patients with pulmonary embolized seeds. CONCLUSION: Embolization of Pd-103 seeds to the lungs after implantation for carcinoma of the prostate is an unusual event. In this study only 0.3% of the seeds implanted migrated to the lungs. Although it was previously thought that pulmonary seed migration mainly occurred on the day of brachytherapy, our experience shows that seeds usually migrated to the lungs after the day of the implant. There were no clinical pulmonary or cardiac effects attributable to embolized seeds in the lungs on routine follow-up. PMID- 9336148 TI - Node-positive prostatic carcinoma. PMID- 9336149 TI - The use of combined radiation therapy and hormonal therapy in the management of lymph node-positive prostate cancer. AB - PURPOSE: To determine the rate of tumor response and patterns of relapse following combined hormonal-radiation therapy of adenocarcinoma of the prostate and to measure the survival in a group of men with tumor metastatic to pelvic lymph nodes. METHODS AND MATERIALS: 66 patients with adenocarcinoma of the prostate with pathologically confirmed pelvic lymph node involvement were treated with combined radiation therapy and hormonal therapy. An additional five patients declined hormonal therapy. The patients treated with combined therapy represented a group with locally advanced disease including 44 patients (67%) with T3 or T4 tumors and 51 patients (80%) had N2 or N3 lymph node metastases. The pelvic lymph nodes were treated to a dose of 45 Gy and the prostate was boosted to a dose of 65 to 71 Gy. Hormonal therapy began up to 2 months before radiation and continued indefinitely. Patients were allowed to select their hormonal therapy and could choose DES (2 patients), orchiectomy (21 patients), LHRH agonist (7 patients) or combined androgen blockade (34 patients). RESULTS: Median follow-up is 49 months (range 12 to 131 months) and 21 patients have been followed for longer than 5 years. There have been 15 recurrences the entire group including three local recurrences in the prostate, seven patients with distant metastases, four patients with biochemical recurrences without clinical evidence of disease, and one patient where the location was unknown. Two of the PSA recurrences occurred in patients who elected to discontinue hormones after less than 3 years of therapy. The overall survival at 5 and 8 years is 94 and 84%, the clinical disease free survival is 85 and 67%, and the biochemical disease-free survival is 78 and 47%. There was no increased toxicity of the combined modality regimen compared to the expected effects of radiation and hormonal therapy. CONCLUSION: Combined hormonal and radiation therapy represents an effective treatment option for patients with adenocarcinoma of the prostate metastatic to pelvic lymph nodes. Combined modality therapy appears to extend the disease-free survival and allow patients to maintain their independent function. PMID- 9336150 TI - Preirradiation PSA predicts biochemical disease-free survival in patients treated with postprostatectomy external beam irradiation. AB - PURPOSE: To assess the clinical outcome and prostate-specific antigen (PSA) response and to determine prognostic factors for biochemical disease-free survival in patients treated with external beam radiotherapy following radical prostatectomy without hormonal therapy. METHODS AND MATERIALS: Forty-eight patients were treated after prostatectomy with radiotherapy between March, 1988 and December, 1993. Seven patients had undetectable PSA (<0.2) and the remainder had detectable PSA at the time of irradiation (overall: median 2.7, range 0 24.9). Nine patients had biopsy proven local recurrence, palpable local disease, or positive preirradiation imaging. No patients received hormonal therapy prior to irradiation. Median follow-up was 55 months. A median dose of 60 Gy (range 58 66) was given to the prostate bed. Survival was analyzed using the life-table method. Actuarial biochemical disease-free survival was the primary endpoint studied. RESULTS: In patients with detectable PSA, 51% had levels return to undetectable after irradiation. The actuarial 5-year freedom from biochemical failure for all patients was 24%. A significant difference in biochemical disease free survival was seen for patients irradiated with preirradiation PSA that was undetectable (p < 0.001), or preirradiation PSA that was < or =2.7 (p = 0.002), vs. preirradiation PSA that was >2.7. Five-year actuarial biochemical disease free survival values were 71, 48, and 0%, respectively, for the three groups. Biochemical disease-free survival was not affected by preoperative PSA level, clinical stage, Gleason's score, pathologic stage, surgical margins, presence of undetectable PSA after surgery, surgery to radiation interval, total dose, or presence of clinically suspicious local disease. Based on digital rectal exam, there were no local failures. CONCLUSION: Biochemical disease-free survival after postprostatectomy radiation is predicted by the PSA at the time of irradiation. Clinical local control is excellent, but distant failure remains a significant problem in this population. The addition of concomitant systemic therapy should be investigated in patients with PSA >2.7. PMID- 9336151 TI - Retinoblastoma protein expression and radiation response in muscle-invasive bladder cancer. AB - PURPOSE: The retinoblastoma protein (pRB) is a key regulator of the G1 cell cycle checkpoint and has been implicated as having a role in G1 arrest and apoptosis induced by radiation damage. In this report we examine the association between pRB expression and radiation response in patients treated between 1960 and 1983 with preoperative radiotherapy (50 Gy in 25 fractions) followed 4-6 weeks later by radical cystectomy. The correlation of pRB to patient outcome and how this relationship is complimentary to that seen with p53 staining status is also described. METHODS AND MATERIALS: Immunohistochemical staining of pRB and p53 in paraffin-embedded tumor sections using WL-1 anti-RB and DO1 anti-p53 antibodies was considered adequate in 98 and 97 pretreatment tumor samples, respectively. There were 46 patients with clinical Stage T2, 28 with Stage T3a, and 24 with Stage T3b disease. The median age was 62 years and follow-up for those living was 85 months. RESULTS: Staining for pRB was negative in 30% of the cases. Correlations were observed between pRB negativity and high pretreatment apoptosis level (p = 0.06), locally advanced clinical stage (p = 0.01), increased clinical to-pathologic downstaging (p = 0.014), and more pathologic complete responses (Path-CRs; p = 0.019). Several other factors were tested and were not associated with pRB status, including p53 expression. RB status was the only pretreatment prognostic factor in the univariate analyses that correlated with downstaging and was independently associated with Path-CR using multivariate logistic regression. Despite these significant relationships, no correlations with patient outcome were observed when the entire cohort was analyzed. Restriction of the analyses to Stage T3b patients, however, revealed that pRB negativity predicted for enhanced distant metastasis freedom (p = 0.006, log rank) and overall survival (p = 0.02). The overexpression of p53 also correlated with distant metastasis freedom and overall survival in Stage T3b patients. Patient outcome was best when RB negative and p53 negative staining were seen. CONCLUSION: Our results indicate that loss of RB function as measured by immunohistochemical staining is the strongest correlate of radiation response thus far recognized. Loss of RB expression also predicted for poor outcome in Stage T3b patients, which appeared to compliment the finding of normal p53 expression. While normal RB protein expression is usually associated with better patient outcome, other series have not examined patients treated with radiotherapy. The absence of pRB may be a useful marker for selecting patients for bladder preservation with radiotherapy, particularly when wild-type p53 is present. PMID- 9336152 TI - The response of human tumors to carbogen breathing, monitored by Gradient Recalled Echo Magnetic Resonance Imaging. AB - PURPOSE: Gradient-Recalled Echo (GRE) Magnetic Resonance Imaging (MRI), which detects changes in blood vessel deoxyhaemoglobin content, has been investigated as a noninvasive monitor of changes in human tumor oxygenation and blood flow, in response to carbogen (95% O2, 5% CO2) breathing. METHODS AND MATERIALS: GRE images (TE = 60 ms, TR = 200 ms, alpha = 40 degrees, 256[2] matrix) were acquired from 31 patients with primary and metastatic disease, prior to and during carbogen breathing. Three patients underwent a follow-up examination after radiotherapy. RESULTS: Seventeen out of 34 tumors showed enhanced image intensity, consistent with an improvement in tumor oxygenation and blood flow, while 11 showed no response; 6 studies were technical failures. In one patient a metastatic node that had eluded orthodox investigation was visualized. A reduction in response was observed in the three patients studied postradiotherapy. CONCLUSION: This method, which can be performed on a standard clinical MRI instrument, provides a noninvasive measurement of tumor response to oxygenation/blood flow modification. In principle, this should enable the clinician to optimize treatment protocols, such as carbogen breathing, for individual radiotherapy patients. PMID- 9336153 TI - The effect of interruptions and prolonged treatment time in radiotherapy for nasopharyngeal carcinoma. AB - PURPOSE: The effect of interruptions and prolonged overall treatment time in radiotherapy for nasopharyngeal carcinoma and the significance of timing of interruption was investigated. METHODS AND MATERIALS: Treatment records of 229 patients treated with continuous course (CC) and 567 patients treated with split course (SC) radiotherapy for nonmetastatic NPC were reviewed. Overall treatment time without inclusion of time for boost was calculated. Treatment that extended 1 week beyond scheduled time was considered prolonged. Outcome in patients who completed treatment "per schedule" were compared with those who had "prolonged" treatment. Because of known patient selection bias between CC and SC, patients on the two schedules were analyzed separately. Multivariate analysis was performed for patients on SC. Total number of days of interruption, age, sex, T and N stage, and the use of boost were tested for the whole SC group. Analysis on the effect of timing of interruption was performed in a subgroup of 223 patients on SC who had a single unplanned interruption. Timing of interruption, either before or after the fourth week for the unplanned interruption, was tested in addition to the other variables in multivariate analysis for this subgroup of SC. RESULTS: Twenty-seven (11.8%) patients on CC and 96 (16.9%) patients on SC had prolonged treatment. Patients on SC who had prolonged treatment had significantly poorer loco-regional control rate and disease free survival when compared with those who completed radiotherapy per schedule (p = 0.0063 and 0.001, respectively, with adjustment for stage). For CC, the effect of prolonged treatment on outcome was not significant. The small number of events for patients on CC probably account for the insignificant finding. The number of days of interruption was confirmed as prognostic factor, independent of T and N stages, for loco-regional control and disease-free survival in multivariate analysis for SC. The hazard rate for loco-regional failure increased by 3.3% for each day of interruption. The timing of interruption, at the beginning or towards end of treatment, did not significantly alter outcome. CONCLUSION: Interruptions and prolonged treatment adversely affect outcome in radiotherapy for NPC and the effect of repopulation was confirmed. Every effort should be made to keep treatment on schedule and interruptions for whatever reasons should be minimized. PMID- 9336154 TI - Volumetric analysis of tumor extent in nasopharyngeal carcinoma and correlation with treatment outcome. AB - PURPOSE: To investigate the variability of tumor volume in nasopharyngeal carcinoma using quantitative measurements of tumor bulk derived from computed tomography, and to study the prognostic value of tumor volume in comparison with other variables. METHODS AND MATERIALS: Two hundred ninety patients with newly diagnosed nasopharyngeal carcinoma were included in the study. The primary tumor volume (PTV) and nodal tumor volume (NTV) were obtained by outlining the tumor contour followed by summation of areas in sequential pretreatment computed tomography axial scans. Total tumor volume (TTV) was obtained by adding the PTV and NTV. All patients had radiotherapy as the primary treatment, 67 patients also received cisplatin-based neoadjuvant chemotheraphy. RESULTS: A large variation in tumor volume was observed, especially in advanced stage disease. The median PTV (cc) in Ho's T1, T2, and T3 disease were: 6.9 (range: 0.9-42.7), 18.8 (1.6 127.9), and 52.4 (3.3-166.8). The median TTV (cc) in Ho's stage I to IV disease were: 7.6 (range: 1.3-42.7), 19.8 (3.2-55.7), 40.7 (4.1-222.7), and 51.1 (3.1 274.7). Patients with a large PTV (>60 cc) were associated with significantly poorer local control (5-year local control rate: 56%) and disease-specific survival (5-year survival rate: 53%). In patients with a small PTV (< or =20 cc), there were no significant differences in local control among different T stages. Large NTV (>30 cc) was associated with significantly higher distant failure rate (5-year distant relapse-free survival rate: 54%) and lower disease-specific survival (5-year survival rate: 40%). In multivariate analysis, only PTV was found to be an independent factor in predicting local control. CONCLUSION: A large variation of tumor volume was present in different T stage disease of nasopharyngeal carcinoma, and PTV represents an independent prognostic factor of local control that appears to be more predictive than Ho's T stage classification. PMID- 9336155 TI - Concurrent platinum-based chemotherapy and hyperfractionated radiotherapy with late intensification in advanced head and neck cancer. AB - PURPOSE: To determine whether a course of hyperfractionated radiation therapy concomitant with escalated radiosensitizing platinum compounds can be administered with acceptable morbidity and achieve a high rate of loco-regional control for Stage III and IV head and neck cancer and whether the patients can be tumor free at the primary site after initial therapy and cured by the additional chemoradiation without radical resection of the primary tumor. METHODS AND MATERIALS: Patients with Stage III/IV head and neck cancer were treated in this multicenter Phase II Study with 1.8 Gy fraction radiotherapy for 2 weeks, with escalation to 1.2 Gy b.i.d. hyperfractionation to 46.8 Gy. Concomitant continuous infusion cisplantinum (CDDP) 20 mg per meter square on day 1 to 4 and 22 to 25 was given. Reassessment by biopsy of primary and nodes was done. Patients with a complete response continued with hyperfractionated radiotherapy to 75.6 Gy with simultaneous carboplatinum (Carbo), 25 mg per meter square b.i.d. for 12 consecutive treatment days. Patients with residual disease at 46.8 Gy required curative surgery. Seventy-four patients were treated at the three institutions; 20 were Stage III and 54 were Stage IV. All patients had daily mouth care, nutritional, and psychosocial support. RESULTS: This regime was well tolerated. Eighty-five percent of toxicities were Grade 1 or 2 and there was only one Grade 4 hematologic toxicity. Late toxicities included xerostomia in 25 patients, dysphasia in 18, and mild speech impediment in 11. Biopsies of primary site were done after the first course of treatment in 59 patients. Neck dissections were performed in 35 patients. Forty-four of 59 (75%) primary sites and 16 of 35 (46%) lymph nodes had pathologically complete response (CR). Of the 74 patients, only 12 required surgical resection of the primary site. Thirty-five of the 50 node positive patients had neck dissections, 16 of these were CRs at surgery. At 4 years (median follow-up of 26 months), disease-specific survival is 63%. The actuarial survival for all patients is 51%. Patients with pathological CR after initial treatment have disease specific survival of 73% at 4 years vs. 48% of patients with partial response (PR) only. CONCLUSION: This study, developed on the basis of radiobiological and cell kinetic precepts, produced results that compare favorably with other reports of management of patients with advanced head and neck cancer. In comparison with our previous study, these results are comparable, not impressively better. The associated morbidity was somewhat worse. PMID- 9336157 TI - Effects of irradiation on the expression of major histocompatibility complex class I antigen and adhesion costimulation molecules ICAM-1 in human cervical cancer. AB - PURPOSE: We initiated studies to analyze the effects of high doses of gamma irradiation on the surface antigen expression of MHC Class I, Class II, and ICAM 1 on human cervical carcinoma cell lines. METHODS AND MATERIALS: The expression of surface antigens (MHC Class I, Class II, and ICAM-1) was evaluated by FACS analysis on two cervical cell lines at different time points, following their exposure to high doses of gamma irradiation (i.e., 25.00, 50.00, and 100.00 Gy). RESULTS: The CaSki and SiHa cervical cancer cells we analyzed in this study expressed variable levels of MHC Class I and ICAM-1 antigens, while Class II surface antigens were not detectable. Whereas irradiation doses of 25.00 Gy were not sufficient to totally block cell replication in both cell lines, exposure to 50.00 or 100.00 Gy was able to completely inhibit cell replication. Range doses from 25.00 to 100.00 Gy significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation but were unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). Importantly, such upregulation was shown to be dose dependent, with higher radiation doses associated with increased antigen expression. Moreover, when the kinetic of this upregulation was studied after 2 and 6 days after irradiation, it was shown to be persistent and lasted until all the cells died. CONCLUSIONS: These findings may partially explain the increased immunogenicity of tumor cells following irradiation and may suggest enhanced immune recognition in tumor tissue in patients receiving radiation therapy. PMID- 9336156 TI - Platelet-derived growth factor (PDGF)-signaling mediates radiation-induced apoptosis in human prostate cancer cells with loss of p53 function. AB - Platelet-derived growth factor (PDGF) signals a diversity of cellular responses in vitro, including cell proliferation, survival, transformation, and chemotaxis. PDGF functions as a "competence factor" to induce a set of early response genes expressed in G1 including p21WAF1/CIP1, a functional mediator of the tumor suppressor gene p53 in G1/S checkpoint. For PDGF-stimulated cells to progress beyond G1 and transit the cell cycle completely, progression factors in serum such as insulin and IGF-1 are required. We have recently shown a novel role of PDGF in inducing apoptosis in growth-arrested murine fibroblasts. The PDGF induced apoptosis is rescued by insulin, suggesting that G1/S checkpoint is a critical determinant for PDGF-induced apoptosis. Because recent studies suggest that radiation-induced signal transduction pathways interact with growth factor mediated signaling pathways, we have investigated whether activation of the PDGF signaling facilitates the radiation-induced apoptosis in the absence of functional p53. For this study we have used the 125-IL cell line, a mutant p53 containing, highly metastatic, and hormone-unresponsive human prostate carcinoma cell line. PDGF signaling is constitutively activated by transfection with a p28v sis expression vector, which was previously shown to activate PDGF alpha- and beta- receptors. Although the basal level of p21WAF1/CIP1 expression and radiation-induced apoptosis were not detectable in control 125-IL cells as would be predicted in mutant p53-containing cells, activation of PDGF-signaling induced expression of p21WAF1/CIP1 and radiation-induced apoptosis. Our study suggests that the level of "competence" growth factors including PDGF may be one of the critical determinants for radiation-induced apoptosis, especially in cells with loss of p53 function at the site of radiotherapy in vivo. PMID- 9336158 TI - Repopulation capacity during fractionated irradiation of squamous cell carcinomas and glioblastomas in vitro. AB - PURPOSE: Determination of clonogenic cell proliferation of three highly malignant squamous cell carcinomas (SCC) and two glioblastoma cell lines during a 20-day course of fractionated irradiation under in vitro conditions. METHODS AND MATERIALS: Tumor cells in exponential growth phase were plated in 24-well plastic flasks and irradiated 24 h after plating with 250 kV x-rays at room temperature. Six fractions with single doses between 0.6 and 9 Gy were administered in 1.67, 5, 10, 15, and 20 days. Colony growth was monitored for at least 60 days after completion of irradiation. Wells with confluent colonies were considered as "recurrences" and wells without colonies as "controlled." The dose required to control 50% of irradiated wells (WCD50) was estimated by a logistic regression for the different overall treatment times. The effective doubling time of clonogenic cells (T[eff]) was determined by a direct fit using the maximum likelihood method. RESULTS: The increase of WCD50 within 18.3 days was highly significant for all tumor cell lines accounting for 7.9 and 12.0 Gy in the two glioblastoma cell lines and for 12.7, 14.0, and 21.7 Gy in the three SCC cell lines. The corresponding T(eff)s were 4.4 and 2.0 days for glioblastoma cell lines and 2.4, 4.2, and 1.8 days for SCC cell lines. Population doubling times (PDT) of untreated tumor cells ranged from 1.0 to 1.9 days, showing no correlation with T(eff)s. T(eff) was significantly longer than PDT in three of five tumor cell lines. No significant differences were observed comparing glioblastomas and SCC. Increase of WCD50 with time did not correlate with T(eff) but with T(eff) InSF2 (surviving fraction at 2 Gy). CONCLUSION: The intrinsic ability of SCC and glioblastoma cells to repopulate during fractionated irradiation could be demonstrated. Repopulation induced dose loss per day depends on T(eff) and intrinsic radiation sensitivity. Proliferation during treatment was decelerated compared to pretreatment PDT in the majority of cell lines. Pretreatment cell kinetics did not predict for tumor cell proliferation during treatment. PMID- 9336159 TI - Pilocarpine, a salivary gland radioprotectant, does not inhibit cytotoxic effect of gamma-radiation on squamous cell carcinoma in vitro. AB - PURPOSE: Pilocarpine, a salivary stimulant, has been shown to protect salivary glands from gamma-radiation-induced damage during the radiotherapy of head and neck tumors. This study was performed to determine whether pilocarpine affects the survival of squamous carcinoma cells, line SCC-25, following gamma-radiation treatment. METHODS AND MATERIALS: The survival of squamous carcinoma tumor cells, line SCC-25, following the exposure of cells to pilocarpine at concentration of 0 100 ng/ml given for 0-1 h prior to radiation at dose of 0-20 Gy was determined by an in vitro colony-formation assay. RESULTS: The survival fractions of SCC-25 cells were identical for the control and pilocarpine-treated samples at all tested conditions. Calculated Do and Dq values did not depend on the presence of pilocarpine and were not affected by the time of incubation prior to irradiation. CONCLUSION: Pilocarpine, at clinically relevant concentrations, given to the SCC 25 cells 1 h prior to or at the time of irradiation did not affect survival of SCC-25 cells in vitro. Pilocarpine does not sensitize or protect these tumor cells from the effects of y-radiation, suggesting that this agent should not compromise the tumoricidal effects of radiotherapy. PMID- 9336160 TI - A technique for accurate planning of stereotactic brain implants prior to head ring fixation. AB - PURPOSE: A two-step procedure is described for accurate planning of stereotactic brain implants prior to head-ring fixation. METHODS AND MATERIALS: Approximately 2 weeks prior to implant a CT scan without the head ring is performed for treatment-planning purposes. An entry point and a reference point, both marked with barium and later tattooed, facilitate planning and permit correlation of the images with a later CT scan. A plan is generated using a conventional treatment planning system to determine the number and activity of I-125 seeds required and the position of each catheter. I-125 seed anisotropy is taken into account by means of a modification to the treatment planning program. On the day of the implant a second CT scan is performed with the head ring affixed to the skull and with the same points marked as in the previous scan. The planned catheter coordinates are then mapped into the coordinate system of the second CT scan by means of a manual translational correction and a computer-calculated rotational correction derived from the reference point coordinates in the two scans. RESULTS: The rotational correction algorithm was verified experimentally in a Rando phantom before it was used clinically. For analysis of the results with individual patients a third CT scan is performed 1 day following the implant and is used for calculating the final dosimetry. CONCLUSION: The technique that is described has two important advantages: 1) the number and activity of seeds required can be accurately determined in advance; and 2) sufficient time is allowed to derive the best possible plan. PMID- 9336161 TI - Surface applicators for high dose rate brachytherapy in AIDS-related Kaposi's sarcoma. AB - PURPOSE: The development of commercially available surface applicators using high dose rate remote afterloading devices has enabled radiotherapy centers to treat selected superficial lesions using a remote afterloading brachytherapy unit. The dosimetric parameters of these applicators, the clinical implementation of this technique, and a review of the initial patient treatment regimes are presented. METHODS AND MATERIALS: A set of six fixed-diameter (1, 2, and 3 cm), tungsten/steel surface applicators is available for use with a single stepping source (Ir-192, 370 GBq) high dose rate afterloader. The source can be positioned either in a parallel or perpendicular orientation to the treatment plane at the center of a conical aperture that sits at an SSD of approximately 15 mm and is used with a 1-mm thick removable plastic cap. The surface dose rates, percent depth dose, and off-axis ratios were measured. A custom-built, ceiling-mounted immobilization device secures the applicator on the surface of the patient's lesion during treatment. RESULTS: Between November 1994, and September 1996, 16 AIDS-related Kaposi's sarcoma patients having a total of 120 lesions have been treated with palliative intent. Treatment sites were distributed between the head and neck, extremity, and torso. Doses ranged from 8 to 20 Gy, with a median dose of 10 Gy delivered in a single fraction. Treatments were well tolerated with minimal skin reaction, except for patients with lesions treated to 20 Gy who developed moderate/severe desquamation. CONCLUSION: Radiotherapy centers equipped with a high dose rate remote afterloading unit may treat small selected surface lesions with commercially available surface applicators. These surface applicators must be used with a protective cap to eliminate electron contamination. The optimal surface dose appears to be either 10 or 15 Gy depending upon the height of the lesion. PMID- 9336162 TI - Regarding, Brenner, Miller and Hall IJROBP 36(4): 805-810, 1996. PMID- 9336163 TI - Pulsed brachytherapy as a substitute for continuous low dose rate: comment on Chen, Huang, Hall, and Brenner's paper. PMID- 9336164 TI - Regarding, Dattoli, Wallner, Sorace, Koval, Cash, Acosta, Brown, Etheridge, Binder, Brunelle, Kirwan, Sanchez, Stein, and Wasserman IJROBP 35(5):875-879; 1996. PMID- 9336165 TI - How well is the net cost of fractionated radiotherapy predicted by radiobiological parameters? PMID- 9336166 TI - Regarding, Allison, Schulsinger, Vongtama, I J ROBP 37:399-403;1997. PMID- 9336167 TI - Molecular dynamics study of free energy profiles for organic cations in gramicidin A channels. AB - The free energy profiles for four organic cations in right-handed single-helix gramicidin A dimers were computed by using umbrella sampling molecular dynamics with CHARMM. Ion-water column translocations were facilitated by using a novel "water-tunnel" approach. The overlapping pieces of free energy profile for adjacent windows were selected from three trajectories that differed in initial ion rotation and were aligned by the method of umbrella potential differences. Neglected long-range electrostatic energies from the bulk water and the bilayer were computed with DelPhi and added to the profile. The approach was corroborated for the formamidinium-guanidinium pair by using perturbation dynamics at axial positions 0, 6, 12, and 15 A from the channel center. The barrier to ethylammonium entry was prohibitive at 21 kcal/mol, whereas for methylammonium it was 5.5 kcal/mol, and the profile was quite flat through the channel, roughly consistent with conductance measurements. The profile for formamidinium was very similar to that of methylammonium. Guanidinium had a high entry barrier (deltaF = +8.6 kcal/mol) and a narrow deep central well (deltaF = -2.6 kcal/mol), qualitatively consistent with predictions from voltage-dependent potassium current block measurements. Its deep central well, contrasting with the flat profile for formamidinium, was verified with perturbation dynamics and was correlated with its high propensity to form hydrogen bonds with the channel at the dimer junction (not shared by the other three cations). Analysis of the ensemble average radial forces on the ions demonstrates that all four ions undergo compressive forces in the channel that are at maximum at the center of the monomer and relieved at the dimer junction, illustrating increased flexibility of the channel walls in the center of the channel. PMID- 9336169 TI - Wetting and capillary condensation as means of protein organization in membranes. AB - Wetting and capillary condensation are thermodynamic phenomena in which the special affinity of interfaces to a thermodynamic phase, relative to the stable bulk phase, leads to the stabilization of a wetting phase at the interfaces. Wetting and capillary condensation are here proposed as mechanisms that in membranes may serve to induce special lipid phases in between integral membrane proteins leading to long-range lipid-mediated joining forces acting between the proteins and hence providing a means of protein organization. The consequences of wetting in terms of protein aggregation and protein clustering are derived both within a simple phenomenological theory as well as within a concrete calculation on a microscopic model of lipid-protein interactions that accounts for the lipid bilayer phase equilibria and direct lipid-protein interactions governed by hydrophobic matching between the lipid bilayer hydrophobic thickness and the length of the hydrophobic membrane domain. The theoretical results are expected to be relevant for optimizing the experimental conditions required for forming protein aggregates and regular protein arrays in membranes. PMID- 9336168 TI - Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles. AB - We previously applied the Poisson-Boltzmann equation to atomic models of phospholipid bilayers and basic peptides to calculate their electrostatic interactions from first principles (Ben-Tal, N., B. Honig, R. M. Peitzsch, G. Denisov, and S. McLaughlan. 1996. Binding of small basic peptides to membranes containing acidic lipids. Theoretical models and experimental results. Biophys. J. 71:561-575). Specifically, we calculated the molar partition coefficient, K (the reciprocal of the lipid concentration at which 1/2 the peptide is bound), of simple basic peptides (e.g., pentalysine) with phospholipid vesicles. The theoretical predictions agreed well with experimental measurements of the binding, but the agreement could have been fortuitous because the structure(s) of these flexible peptides is not known. Here we use the same theoretical approach to calculate the membrane binding of two small proteins of known structure: charybdotoxin (CTx) and iberiotoxin (IbTx); we also measure the binding of these proteins to phospholipid vesicles. The theoretical model describes accurately the dependence of K on the ionic strength and mol % acidic lipid in the membrane for both CTx (net charge +4) and IbTx (net charge +2). For example, the theory correctly predicts that the value of K for the binding of CTx to a membrane containing 33% acidic lipid should decrease by a factor of 10(5) when the salt concentration increases from 10 to 200 mM. We discuss the limitations of the theoretical approach and also consider a simple extension of the theory that incorporates nonpolar interactions. PMID- 9336170 TI - Configurational transitions in Fourier series-represented DNA supercoils. AB - A new Fourier series representation of supercoiled DNA is employed in Langevin dynamics simulations to study large-scale configurational motions of intermediate length chains. The polymer is modeled as an ideal elastic rod subject to long range van der Waals' interactions. The van der Waals' term prevents the self contact of distant chain segments and also mimics attractive forces thought to stabilize the association of closely spaced charged rods. The finite Fourier series-derived polymer formulation is an alternative to the piecewise B-spline curves used in past work to describe the motion of smoothly deformed supercoiled DNA in terms of a limited number of independent variables. This study focuses on two large-scale configurational events: the interconversion between circular and figure-8 forms at a relatively low level of supercoiling, and the transformation between branched and interwound structures at a higher superhelical density. PMID- 9336171 TI - Molecular polarity in tropomyosin-troponin T co-crystals. AB - New features of the structure and interactions of troponin T and tropomyosin have been revealed by electron microscopy of so-called double-diamond co-crystals. These co-crystals were formed using rabbit alpha2 tropomyosin complexed with troponin T from either skeletal or cardiac muscle, which have different lengths in the amino-terminal region, as well as a bacterially expressed skeletal muscle troponin T fragment of 190 residues that lacks the amino-terminal region. Differences in the images of the co-crystals have allowed us to establish the polarities of both the troponin T subunit and tropomyosin in the projected lattice. Moreover, in agreement with their sequences, the amino-terminal region of a bovine cardiac muscle troponin T isoform appears to be longer than that from the rabbit skeletal muscle troponin T isoform and to span more of the amino terminus of tropomyosin at the head-to-tail filament joints. Images of crystals tilted relative to the electron beam also reveal the supercoiling of the tropomyosin filaments in this lattice. Based on these results, a three dimensional model of the double-diamond lattice has been constructed. PMID- 9336172 TI - Liquid crystalline ordering of nucleosome core particles under macromolecular crowding conditions: evidence for a discotic columnar hexagonal phase. AB - Macromolecular crowding conditions occurring inside the cell nucleus were reproduced experimentally with solutions of mononucleosome core particles to study their supramolecular organization. We report here that under these conditions, and over a large range of monovalent salt concentrations, mononucleosome core particles self-assemble to form a discotic liquid crystalline phase characterized in polarizing and freeze-fracture electron microscopy. Mononucleosomes are stacked on each other to form columns, which are themselves closely packed into an hexagonal array. The nucleosome concentration, estimated from the network parameters, falls in the range of values measured in cell nuclei. We suggest that these concentrated solutions, although their organization cannot be immediately compared to the organization of chromatin in vivo, may be used to investigate the nucleosome-nucleosome interactions. Furthermore, this approach may be complexified to take into account the complexity of the eucaryotic chromatin. PMID- 9336174 TI - Electroporation-induced formation of individual calcium entry sites in the cell body and processes of adherent cells. AB - Electroporation is a widely used method for introducing macromolecules into cells. We developed an electroporation device that requires only 1 microl of sample to load adherent cells in a 10-mm2 surface area while retaining greater than 90% cell survivability. To better understand this device, field-induced permeabilization of adherent rat basophilic leukemia and neocortical neuroblastoma cells was investigated by using fluorescent calcium and voltage indicators. Rectangular field pulses led to the formation of only a few calcium entry sites, preferentially in the hyperpolarized parts of the cell body and processes. Individual entry sites were formed at the same locations when field pulses were repeated. Before calcium entry, a partial breakdown of the membrane potential was observed in both polar regions. Based on our results, a model is proposed for the formation and closure of macromolecule entry sites in adherent cells. First, the rapid formation of a large number of small pores leads to a partial membrane potential breakdown in both polar regions of the cell. Second, over tens of milliseconds, a few entry sites for macromolecules are formed, preferentially in the hyperpolarized part of cell body and processes, at locations defined by the local membrane structure. These entry sites reseal on a time scale of 50 ms to several seconds, with residual small pores remaining open for several minutes. PMID- 9336173 TI - Proximity oscillations of complement type 4 (alphaX beta2) and urokinase receptors on migrating neutrophils. AB - Migrating neutrophils utilize beta2 integrins for substrate attachment and urokinase receptors (uPAR) to focus pericellular proteolysis. Our studies show that CR3 associates with uPAR on resting cells, whereas uPAR associates with CR4 at lamellipodia of migrating cells. Using resonance energy transfer (RET) microscopy, we show that the molecular proximity between CR4 and uPAR oscillates on migrating cells, thus suggesting that CR4 molecules periodically bind/release uPAR. Cell contact with fibrinogen, endothelial cells, chemotactic factors and indomethacin, and treatment with sub-optimal doses of signal transduction inhibitors, affect the oscillations' period, amplitude, and/or waveform. The oscillations were indistinguishable in period and 180 degrees out-of-phase with cytosolic NAD(P)H autofluorescence oscillations. Thus, CR4 and CR3 identify a neutrophil's axis of migration and CR4 may restrain uPAR at lamellipodia. Oscillations in signal transduction and energy metabolism may coordinate cell adherence, local proteolysis, oxidant release, actin assembly, and cell extension. PMID- 9336175 TI - Dynamics of pore growth in membranes and membrane stability. AB - Pores can form and grow in biomembranes because of factors such as thermal fluctuation, transmembrane electrical potential, and cellular environment. We propose a new statistical physics model of the pore growth treated as a non Markovian stochastic process, with a free energy barrier and memory friction from the membrane matrix treated as a quasi-two-dimensional viscoelastic and dielectric fluid continuum. On the basis of the modern theory of activated barrier crossing, an analytical expression for membrane lifetime and the phase diagram for membrane stability are obtained. The memory effect due to membrane viscoelasticity and the elasticity due to cytoskeletal network are found to induce sharp transitions to membrane stability against pore growth and compete with other factors to manifest rich dynamic transitions over the membrane lifetime. PMID- 9336176 TI - A molecular theory for nonohmicity of the ion leak across the lipid-bilayer membrane. AB - The current-voltage relationship of ion leak (i.e., ion transport involving neither special channels nor carriers) across the lipid-bilayer membrane has been observed to be log-linear above the ohmic regime. The coefficient of the linear term has been found to be universal for membranes and penetrants examined. This universality has been explained in terms of diffusion in an external field, where the ion position is described as a Markovian process. Such a diffusion picture can be questioned, however. It is also probable that a leaking ion gets over the potential barrier before experiencing sufficient random collision in the membrane, considering that each ion is surrounded with long lipid molecules aligned almost unidirectionally. As an alternative, we discuss this ion leak in terms of velocity distribution of the ions entering the membrane and density fluctuation of the lipids. We conclude that we can explain the universality without resorting to the diffusion picture. PMID- 9336177 TI - Probabilistic secretion of quanta and the synaptosecretosome hypothesis: evoked release at active zones of varicosities, boutons, and endplates. AB - A quantum of transmitter may be released upon the arrival of a nerve impulse if the influx of calcium ions through a nearby voltage-dependent calcium channel is sufficient to activate the vesicle-associated calcium sensor protein that triggers exocytosis. A synaptic vesicle, together with its calcium sensor protein, is often found complexed with the calcium channel in active zones to form what will be called a "synaptosecretosome." In the present work, a stochastic analysis is given of the conditions under which a quantum is released from the synaptosecretosome by a nerve impulse. The theoretical treatment considers the rise of calcium at the synaptosecretosome after the stochastic opening of a calcium channel at some time during the impulse, followed by the stochastic binding of calcium to the vesicle-associated protein and the probability of this leading to exocytosis. This allows determination of the probabilities that an impulse will release 0, 1, 2,... quanta from an active zone, whether this is in a varicosity, a bouton, or a motor endplate. A number of experimental observations of the release of transmitter at the active zones of sympathetic varicosities and boutons as well as somatic motor endplates are described by this analysis. These include the likelihood of the secretion of only one quantum at an active zone of endplates and of more than one quantum at an active zone of a sympathetic varicosity. The fourth-power relationship between the probability of transmitter release at the active zones of sympathetic varicosities and motor endplates and the external calcium concentration is also explained by this approach. So, too, is the fact that the time course of the increased rate of quantal secretion from a somatic active zone after an impulse is invariant with changes in the amount of calcium that enters through its calcium channel, whether due to changes consequent on the actions of autoreceptor agents such as adenosine or to facilitation. The increased probability of quantal release that occurs during F1 facilitation at the active zones of motor endplates and sympathetic boutons is predicted by the residual binding of calcium to a high affinity site on the vesicle-associated protein. The concept of the stochastic operation of a synaptosecretosome can accommodate most phenomena involving the release of transmitter quanta at these synapses. PMID- 9336178 TI - Computation of molecular electrostatics with boundary element methods. AB - In continuum approaches to molecular electrostatics, the boundary element method (BEM) can provide accurate solutions to the Poisson-Boltzmann equation. However, the numerical aspects of this method pose significant problems. We describe our approach, applying an alpha shape-based method to generate a high-quality mesh, which represents the shape and topology of the molecule precisely. We also describe an analytical method for mapping points from the planar mesh to their exact locations on the surface of the molecule. We demonstrate that derivative boundary integral formulation has numerical advantages over the nonderivative formulation: the well-conditioned influence matrix can be maintained without deterioration of the condition number when the number of the mesh elements scales up. Singular integrand kernels are characteristics of the BEM. Their accurate integration is an important issue. We describe variable transformations that allow accurate numerical integration. The latter is the only plausible integral evaluation method when using curve-shaped boundary elements. PMID- 9336179 TI - Multilamellar structures of DNA complexes with cationic liposomes. AB - Studies of DNA complexes with cationic liposomes are prompted by the search for nonviral DNA carriers for gene therapy. Recent experiments have identified a stable multilamellar phase in which ordered smectic layers of DNA alternate with cationic bilayers. In this paper we identify the forces governing DNA adsorption on cationic lamellae, including a membrane-induced attraction between the adsorbed DNA. Calculating the DNA interhelical spacing as a function of system composition, the model successfully explains recent surprising observations. PMID- 9336180 TI - Prediction of solution structures of the Ca2+-bound gamma-carboxyglutamic acid domains of protein S and homolog growth arrest specific protein 6: use of the particle mesh Ewald method. AB - The solution structures of the N-terminal domains of protein S, a plasma vitamin K-dependent glycoprotein, and its homolog growth arrest specific protein 6 (Gas6) were predicted by molecular dynamics computer simulations. The initial structures were based on the x-ray crystallographic structure of the corresponding region of bovine prothrombin fragment 1. The subsequent molecular dynamics trajectories were calculated using the second-generation AMBER force field. The long-range electrostatic forces were evaluated by the particle mesh Ewald method. The structures that stabilized over a 400-ps time interval were compared with the corresponding region of the simulated solution structure of bovine prothrombin fragment 1. Structural properties of the gamma-carboxyglutamic acid (Gla) domains obtained from simulations and calcium binding were found to be conserved for all three proteins. Analysis of the predicted solution structure of the Gla domain of Gas6 suggests that this domain should bind with negatively charged phospholipid surfaces analogous to bovine prothrombin fragment 1 and protein S. PMID- 9336181 TI - Intracellular Ca2+ inactivates L-type Ca2+ channels with a Hill coefficient of approximately 1 and an inhibition constant of approximately 4 microM by reducing channel's open probability. AB - The patch-clamp technique was used to characterize the mechanism of Ca2+-induced inactivation of cardiac L-type Ca2+ channel alpha(1C-a) + beta3 subunits stably expressed in CHO cells. Single Ca2+ channel activity was monitored with 96 mM Ba2+ as charge carrier in the presence of 2.5 microM (-)BAYK 8644 and calpastatin plus ATP. This enabled stabilization of channel activity in the inside-out patch and allowed for application of steady-state Ca2+ concentrations to the intracellular face of excised membrane patches in an attempt to provoke Ca2+ induced inactivation. Inactivation was found to occur specifically with Ca2+ since it was not observed upon application of Ba2+. Ca2+-dependent inhibition of mean Ca2+ channel activity was characterized by a Hill coefficient close to 1. Ca2+ binding to open and closed states of the channel obtained during depolarization apparently occurred with similar affinity yielding half-maximal inhibition of Ca2+ channel activity at approximately 4 microM. This inhibition manifested predominantly in a reduction of the channel's open probability whereas availability remained almost unchanged. The reduction in open probability was achieved by an increase in first latencies and a decrease in channel opening frequency as well as channel open times. At high (12-28 microM) Ca2+ concentrations, 72% of inhibition occurred due to a stabilization of the closed state and the remaining 28% by a destabilization of the open state. Our results suggest that binding of one calcium ion to a regulatory domain induces a complex alteration in the kinetic properties of the Ca2+ channel and support the idea of a single EF hand motif as the relevant Ca2+ binding site on the alpha1 subunit. PMID- 9336182 TI - Effect of high hydrostatic pressure on the BK channel in bovine chromaffin cells. AB - The activity of the BK channel of bovine chromaffin cells was studied at high hydrostatic pressure, using inside-out patches in symmetrical KCl solution, Ca2+ free and at V(H) = -60 to -40 mV. Pressure increased the probability of channels being open (900 atm increasing the probability 30-fold), and it increased the minimum number of channels apparent in the patches. The pressure activation of the channel was reversed on decompression. Channel conductance was unaffected. It was shown that pressure did not act by raising the temperature, or by affecting [Ca] or pH, or the order of the membrane bilayer, and it was concluded that pressure most likely acted directly on the channel proteins and/or their modulating reactions. PMID- 9336183 TI - Pore residues critical for mu-CTX binding to rat skeletal muscle Na+ channels revealed by cysteine mutagenesis. AB - We have studied mu-conotoxin (mu-CTX) block of rat skeletal muscle sodium channel (rSkM1) currents in which single amino acids within the pore (P-loop) were substituted with cysteine. Among 17 cysteine mutants expressed in Xenopus oocytes, 7 showed significant alterations in sensitivity to mu-CTX compared to wild-type rSkM1 channel (IC50 = 17.5 +/- 2.8 nM). E758C and D1241C were less sensitive to mu-CTX block (IC50 = 220 +/- 39 nM and 112 +/- 24 nM, respectively), whereas the tryptophan mutants W402C, W1239C, and W1531C showed enhanced mu-CTX sensitivity (IC50 = 1.9 +/- 0.1, 4.9 +/- 0.9, and 5.5 +/- 0.4 nM, respectively). D400C and Y401C also showed statistically significant yet modest (approximately twofold) changes in sensitivity to mu-CTX block compared to WT (p < 0.05). Application of the negatively charged, sulfhydryl-reactive compound methanethiosulfonate-ethylsulfonate (MTSES) enhanced the toxin sensitivity of D1241C (IC50 = 46.3 +/- 12 nM) while having little effect on E758C mutant channels (IC50 = 199.8 +/- 21.8 nM). On the other hand, the positively charged methanethiosulfonate-ethylammonium (MTSEA) completely abolished the mu-CTX sensitivity of E758C (IC50 > 1 microM) and increased the IC50 of D1241C by about threefold. Applications of MTSEA, MTSES, and the neutral MTSBN (benzyl methanethiosulfonate) to the tryptophan-to-cysteine mutants partially or fully restored the wild-type mu-CTX sensitivity, suggesting that the bulkiness of the tryptophan's indole group is a determinant of toxin binding. In support of this suggestion, the blocking IC50 of W1531A (7.5 +/- 1.3 nM) was similar to W1531C, whereas W1531Y showed reduced toxin sensitivity (14.6 +/- 3.5 nM) similar to that of the wild-type channel. Our results demonstrate that charge at positions 758 and 1241 are important for mu-CTX toxin binding and further suggest that the tryptophan residues within the pore in domains I, III, and IV negatively influence toxin-channel interaction. PMID- 9336184 TI - Interaction between the sodium channel inactivation linker and domain III S4-S5. AB - The III-IV linker (L(III-IV)) of the rat brain sodium channel is critical for fast inactivation, possibly forming a fast inactivation particle. Inactivation can be disrupted by mutation of a conserved alanine at position 1329 in the S4-S5 loop of domain III. Combination of a charged mutation at 1329 with a compensatory (opposite) charge mutation at position 1489 in L(III-IV) partially restores inactivation of the channel. The compensatory charge mutant channel has a single channel mean open time that is similar to that of the wild-type channel and is approximately 50 times shorter than that of the L(III-IV) mutant channel. The results of thermodynamic cycle analysis indicate that the mutations in domain III S4-S5 and L(III-IV) have a coupling energy of 2.8 kcal/mol, indicating that the two mutations act interdependently. These data suggest that L(III-IV) interacts directly with A1329, which may form part of the docking site if L(III-IV) is a fast inactivation particle. PMID- 9336185 TI - Defective fast inactivation recovery and deactivation account for sodium channel myotonia in the I1160V mutant. AB - The skeletal muscle sodium channel mutant I1160V cosegregates with a disease phenotype producing myotonic discharges (observed as muscle stiffness) that are worsened by elevated K+ levels but unaffected by cooling. The I1160V alpha subunit was co-expressed with the beta1-subunit in Xenopus oocytes. An electrophysiological characterization was undertaken to examine the underlying biophysical characteristics imposed by this mutation. Two abnormalities were found. 1) The voltage dependence of steady-state fast inactivation was reduced in I1160V, which resulted in faster rates of closed-state fast inactivation onset and recovery in I1160V compared with wild-type channels. 2) The rates of deactivation were slower in I1160V than in wild-type channels. Using a computer simulated model, the combination of both defects elicited myotonic runs under conditions of elevated K+, consistent with the observed phenotype of the mutant. PMID- 9336186 TI - Single-channel properties of the recombinant skeletal muscle Ca2+ release channel (ryanodine receptor). AB - We report transient expression of a full-length cDNA encoding the Ca2+ release channel of rabbit skeletal muscle sarcoplasmic reticulum (ryanodine receptor) in HEK-293 cells. The single-channel properties of the 3-[(3 cholamidopropyl)dimethylammonio]-1-propane sulfonate-solubilized and sucrose gradient-purified recombinant Ca2+ release channels were investigated by using single-channel recordings in planar lipid bilayers. The recombinant Ca2+ release channel exhibited a K+ conductance of 780 pS when symmetrical 250 mM KCl was used as the conducting ion and a Ca2+ conductance of 116 pS in 50 mM luminal Ca2+. Opening events of the recombinant channels were brief, with an open time constant of approximately 0.22 ms. The recombinant Ca2+ release channel was more permeable to Ca2+ than to K+, with a pCa2+/pK+ ratio of 6.8. The response of the recombinant Ca2+ release channel to various concentrations of Ca2+ was biphasic, with the channel being activated by micromolar Ca2+ and inhibited by millimolar Ca2+. The recombinant channels were activated by ATP and caffeine, inhibited by Mg2+ and ruthenium red, and modified by ryanodine. Most recombinant channels were asymmetrically blocked, conducting current unidirectionally from the luminal to the cytoplasmic side of the channel. These data demonstrate that the properties of recombinant Ca2+ release channel expressed in HEK-293 cells are very similar, if not identical, to those of the native channel. PMID- 9336187 TI - Reduced inhibitory effect of Mg2+ on ryanodine receptor-Ca2+ release channels in malignant hyperthermia. AB - Malignant hyperthermia (MH) is a potentially fatal, inherited skeletal muscle disorder in humans and pigs that is caused by abnormal regulation of Ca2+ release from the sarcoplasmic reticulum (SR). MH in pigs is associated with a single mutation (Arg615Cys) in the SR ryanodine receptor (RyR) Ca2+ release channel. The way in which this mutation leads to excessive Ca2+ release is not known and is examined here. Single RyR channels from normal and MH-susceptible (MHS) pigs were examined in artificial lipid bilayers. High cytoplasmic (cis) concentrations of either Ca2+ or Mg2+ (>100 microM) inhibited channel opening less in MHS RyRs than in normal RyRs. This difference was more prominent at lower ionic strength (100 mM versus 250 mM). In 100 mM cis Cs+, half-maximum inhibition of activity occurred at approximately 100 microM Mg2+ in normal RyRs and at approximately 300 microM Mg2+ in MHS RyRs, with an average Hill coefficient of approximately 2 in both cases. The level of Mg2+ inhibition was not appreciably different in the presence of either 1 or 50 microM activating Ca2+, showing that it was not substantially influenced by competition between Mg2+ and Ca2+ for the Ca2+ activation site. Even though the absolute inhibitory levels varied widely between channels and conditions, the inhibitory effects of Ca2+ and Mg2+ were virtually identical for the same conditions in any given channel, indicating that the two cations act at the same low-affinity inhibitory site. It seems likely that at the cytoplasmic [Mg2+] in vivo (approximately 1 mM), this Ca2+/Mg2+-inhibitory site will be close to fully saturated with Mg2+ in normal RyRs, but less fully saturated in MHS RyRs. Therefore MHS RyRs should be more sensitive to any activating stimulus, which would readily account for the development of an MH episode. PMID- 9336188 TI - Estimation of the pore size of the large-conductance mechanosensitive ion channel of Escherichia coli. AB - The open channel diameter of Escherichia coli recombinant large-conductance mechanosensitive ion channels (MscL) was estimated using the model of Hille (Hille, B. 1968. Pharmacological modifications of the sodium channels of frog nerve. J. Gen. Physiol. 51:199-219) that relates the pore size to conductance. Based on the MscL conductance of 3.8 nS, and assumed pore lengths, a channel diameter of 34 to 46 A was calculated. To estimate the pore size experimentally, the effect of large organic ions on the conductance of MscL was examined. Poly-L lysines (PLLs) with a diameter of 37 A or larger significantly reduced channel conductance, whereas spermine (approximately 15 A), PLL19 (approximately 25 A) and 1,1'-bis-(3-(1'-methyl-(4,4'-bipyridinium)-1-yl)-propyl)-4,4'-b ipyridinium (approximately 30 A) had no effect. The smaller organic ions putrescine, cadaverine, spermine, and succinate all permeated the channel. We conclude that the open pore diameter of the MscL is approximately 40 A, indicating that the MscL has one of the largest channel pores yet described. This channel diameter is consistent with the proposed homohexameric model of the MscL. PMID- 9336189 TI - Diffusion behavior of lipid vesicles in entangled polymer solutions. AB - Dynamic light scattering was used to follow the tracer diffusion of phospholipid/cholesterol vesicles in aqueous polyacrylamide solutions and compared with the diffusive behavior of polystyrene (PS) latex spheres of comparable diameters. Over the range of the matrix concentration examined (Cp = 0.1-10 mg/ml), the diffusivities of the PS spheres and the large multilamellar vesicles exhibited the Stokes-Einstein (SE) relation, while the diffusivity of the unilamellar vesicles did not follow the increase of the solution's viscosity caused by the presence of the matrix molecules. The difference between the diffusion behaviors of unilamellar vesicles and hard PS spheres of similar size is possibly due to the flexibility of the lipid bilayer of the vesicles. The unilamellar vesicles are capable of changing their shape to move through the entangled polymer solution so that the hindrance to their diffusion due to the presence of the polymer chains is reduced, while the rigid PS spheres have little flexibility and they encounter greater resistance. The multilamellar vesicles are less flexible, thus their diffusion is similar to the hard PS spheres of similar diameter. PMID- 9336191 TI - Membrane lateral compressibility determined by NMR and x-ray diffraction: effect of acyl chain polyunsaturation. AB - The elastic area compressibility modulus, Ka, of lamellar liquid crystalline bilayers was determined by a new experimental approach using 2H-NMR order parameters of lipid hydrocarbon chains together with lamellar repeat spacings measured by x-ray diffraction. The combination of NMR and x-ray techniques yields accurate determination of lateral area per lipid molecule. Samples of saturated, monounsaturated, and polyunsaturated phospholipids were equilibrated with polyethylene glycol (PEG) 20,000 solutions in water at concentrations from 0 to 55 wt % PEG at 30 degrees C. This procedure is equivalent to applying 0 to 8 dyn/cm lateral pressure to the bilayers. The resulting reductions in area per lipid were measured with a resolution of +/-0.2 A2 and the fractional area decrease was proportional to applied lateral pressure. For 1,2-dimyristoyl(d54) sn-glycero-3-phosphocholine, 1-stearoyl(d35)-2-oleoyl-sn-glycero-3-phosphocholine (SOPC-d35), and 1-stearoyl(d35)-2-docosahexaenoyl-sn-glycero-3-phosphocholine (SDPC-d35) cross-sectional areas per molecule in excess water of 59.5, 61.4, and 69.2 A2 and bilayer elastic area compressibility moduli of 141, 221, and 121 dyn/cm were determined, respectively. Combining NMR and x-ray results enables the determination of compressibility differences between saturated and unsaturated hydrocarbon chains. In mixed-chain SOPC-d35 both chains have similar compressibility moduli; however, in mixed-chain polyunsaturated SDPC-d35, the saturated stearic acid chain appears to be far less compressible than the polyunsaturated docosahexaenoic acid chain. PMID- 9336190 TI - Molecular sorting of lipids by bacteriorhodopsin in dilauroylphosphatidylcholine/distearoylphosphatidylcholine lipid bilayers. AB - A combined experimental and theoretical study is performed on binary dilauroylphosphatidylcholine/distearoylphosphatidylcholine (DLPC/DSPC) lipid bilayer membranes incorporating bacteriorhodopsin (BR). The system is designed to investigate the possibility that BR, via a hydrophobic matching principle related to the difference in lipid bilayer hydrophobic thickness and protein hydrophobic length, can perform molecular sorting of the lipids at the lipid-protein interface, leading to lipid specificity/selectivity that is controlled solely by physical factors. The study takes advantage of the strongly nonideal mixing behavior of the DLPC/DSPC mixture and the fact that the average lipid acyl-chain length is strongly dependent on temperature, particularly in the main phase transition region. The experiments are based on fluorescence energy transfer techniques using specifically designed lipid analogs that can probe the lipid protein interface. The theoretical calculations exploit a microscopic molecular interaction model that embodies the hydrophobic matching as a key parameter. At low temperatures, in the gel-gel coexistence region, experimental and theoretical data consistently indicate that BR is associated with the short-chain lipid DLPC. At moderate temperatures, in the fluid-gel coexistence region, BR remains in the fluid phase, which is mainly composed of short-chain lipid DLPC, but is enriched at the interface between the fluid and gel domains. At high temperatures, in the fluid phase, BR stays in the mixed lipid phase, and the theoretical data suggest a preference of the protein for the long-chain DSPC molecules at the expense of the short-chain DLPC molecules. The combined results of the experiments and the calculations provide evidence that a molecular sorting principle is active because of hydrophobic matching and that BR exhibits physical lipid selectivity. The results are discussed in the general context of membrane organization and compartmentalization and in terms of nanometer-scale lipid-domain formation. PMID- 9336192 TI - Evidence for superlattice arrangements in fluid phosphatidylcholine/phosphatidylethanolamine bilayers. AB - Recently, evidence for cholesterol and phosphatidylcholine (PC) molecules to adapt superlattice arrangements in fluid lipid bilayers has been presented. Whether superlattice arrangements exist in other biologically relevant lipid membranes, such as phosphatidylethanolamine (PE)/PC, is still speculative. In this study, we have examined the physical properties of fluid 1-palmitoyl-2 oleoyl-PC (POPC) and 1-palmitoyl-2-oleoyl-PE (POPE) binary mixtures as a function of the POPE mole fraction (X(PE)) using fluorescence and Fourier transform infrared spectroscopy. At 30 degrees C, i.e., above the Tm of POPE and POPC, deviations, or dips, as well as local data scattering in the excimer-to-monomer fluorescence intensity ratio of intramolecular excimer forming dipyrenylphosphatidylcholine probe in POPE/POPC mixtures were detected at X(PE) approximately 0.04, 0.11, 0.16, 0.26, 0.33, 0.51, 0.66, 0.75, 0.82, 0.91, and 0.94. The above critical values of X(PE) coincide (within +/-0.03) with the critical mole fractions X(HX,PE) or X(R,PE) predicted by a headgroup superlattice model, which assumes that the lipid headgroups form hexagonal or rectangular superlattice, respectively, in the bilayer. Other spectroscopic data, generalized polarization of Laurdan and infrared carbonyl and phosphate stretching frequency, were also collected. Similar agreements between some of the observed critical values of X(PE) from these data and the X(HX,PE) or X(R,PE) values were also found. However, all techniques yielded critical values of X(PE) (e.g., 0.42 and 0.58) that cannot be explained by the present headgroup superlattice model. The effective cross-sectional area of the PE headgroup is smaller than that of the acyl chains. Hence, the relief of "packing frustration" of PE in the presence of PC (larger headgroup than PE) may be one of the major mechanisms in driving the PE and PC components to superlattice-like lateral distributions in the bilayer. We propose that headgroup superlattices may play a significant role in the regulation of membrane lipid compositions in cells. PMID- 9336193 TI - Permeabilization and fusion of uncharged lipid vesicles induced by the HIV-1 fusion peptide adopting an extended conformation: dose and sequence effects. AB - The peptide HIV(arg), corresponding to a sequence of 23 amino acid residues at the N-terminus of HIV-1 gp41 (LAV1a strain), has the capacity to destabilize negatively charged large unilamellar vesicles. As revealed by infrared spectroscopy, the peptide associated with those vesicles showed conformational polymorphism: in the absence of cations the main structure was a pore-forming alpha-helix, whereas in the presence of Ca2+ the conformation switched to a fusogenic, predominantly extended beta-type structure. Here we show that an extended structure can also be involved in electrically neutral vesicle destabilization induced by the HIV-1 fusion peptide when it binds the vesicle from the aqueous phase. In the absence of cations, neutral liposomes composed of phosphatidylcholine, phosphatidylethanolamine, and cholesterol (molar ratio 1:1:1) selected for an extended structure that became fusogenic in a dose dependent fashion. At subfusogenic doses this structure caused the release of trapped 8-aminonaphtalene-1,3,6-trisulfonic acid sodium salt/p xylenebis(pyridinium)bromide from liposomes, indicating the existence of a peptide-mediated membrane destabilizing process before and independent of the development of fusion. When compared to HIV(arg), the fusion activity of HIV(ala) (bearing the R22 --> A substitution) was reduced by 70%. Fusogenicity was completely abolished when a second substitution (V2 --> E) was included to generate HIV(ala-E2), a sequence representing the N-terminus of an inactive gp41. However, the three sequences associated with vesicles to the same extent, and the three adopted a similar extended structure in the membrane. Whereas 1-(4 trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene emission anisotropy was unaffected by the three peptides, DPH emission anisotropy in membranes was increased only by the fusogenic sequences. Taken together, our observations strongly argue that it is not an alpha-helical but an extended structure adopted by the HIV-1 fusion peptide what actively destabilizes cholesterol-containing, electrically neutral membranes. Moreover, membrane destabilization is modulated by the amino acid sequence in the extended structure. The effect displayed by the aforementioned V2 --> E substitution suggests that the fusion process described here could be reflecting a physiologically relevant phenomenon. PMID- 9336194 TI - Cytoskeletal protein binding kinetics at planar phospholipid membranes. AB - It has been hypothesized that nonspecific reversible binding of cytoskeletal proteins to lipids in cells may guide their binding to integral membrane anchor proteins. In a model system, we measured desorption rates k(off) (off-rates) of the erythrocyte cytoskeletal proteins spectrin and protein 4.1 labeled with carboxyfluorescein (CF), at two different compositions of planar phospholipid membranes (supported on glass), using the total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP) technique. The lipid membranes consisted of either pure phosphatidylcholine (PC) or a 3:1 mixture of PC with phosphatidylserine (PS). In general, the off-rates were not single exponentials and were fit to a combination of fast, slow, and irreversible fractions, reported both separately and as a weighted average. By a variation of TIR/FRAP, we also measured equilibrium affinities (the ratio of surface-bound to bulk protein concentration) and thereby calculated on-rates, k(on). The average off-rate of CF 4.1 from PC/PS (approximately 0.008/s) is much slower than that from pure PC (approximately 1.7/s). Despite the consequent increase in equilibrium affinity at PC/PS, the on-rate at PC/PS is also substantially decreased (by a factor of 40) relative to that at pure PC. The simultaneous presence of (unlabeled) spectrin tends to substantially decrease the on-rate (and the affinity) of CF-4.1 at both membrane types. Similar experiments for CF-spectrin alone showed much less sensitivity to membrane type and generally faster off-rates than those exhibited by CF-4.1. However, when mixed with (unlabeled) 4.1, both the on-rate and off rate of CF-spectrin decreased drastically at PC/PS (but not PC), leading to a somewhat increased affinity. Clearly, changes in affinity often involve countervailing changes in both on-rates and off-rates. In many of these studies, the effect of varying ionic strength and bulk concentrations was examined; it appears that the binding is an electrostatic attraction and is far from saturation at the concentrations employed. These results and the techniques implemented carry general implications for understanding the functional role of nonspecific protein binding to cellular lipid membranes. PMID- 9336195 TI - Kinetics of contractile activation in voltage clamped frog skeletal muscle fibers. AB - Excitation-contraction coupling events leading to the onset of contraction were studied in single skeletal frog muscle fibers. This entailed the simultaneous measurement of the changes in intracellular calcium concentration using antipyrylazo III and fura-2, isometric force, and clamp voltage in a modified single vaseline gap chamber for the first time. The calcium transients were incorporated into an analysis of calcium binding to regulatory sites of troponin C (TnC) that permitted both a linear and a cooperative interaction. The analysis assumed that the onset of mechanical activation corresponds with a particular TnC saturation with calcium setting constraints for the calcium binding parameters of the regulatory sites. Using a simple model that successfully reproduced both the time course and the relative amplitudes of the measured isometric force transients over a wide membrane potential range, k(off) of TnC was calculated to be 78 s(-1) for the cooperative model at 10 degrees C. Together with the above constraints this gave a dissociation constant of 8.8 +/- 2.5 microM and a relative TnC saturation at the threshold (Sth) that would cause just detectable movement of 0.17 +/- 0.03 (n = 13; mean +/- SE). The predictions were found to be independent of the history of calcium binding to the regulatory sites. The observed delay between reaching Sth and the onset of fiber movement (8.7 +/- 1.0 ms; mean +/- SE, n = 37; from seven fibers) was independent of the membrane potential giving an upper estimate for the delay in myofilament activation. We thus emerge with quantitative values for the calcium binding to the regulatory sites on TnC under maintained structural conditions close to those in vivo. PMID- 9336196 TI - Mechanics of single kinesin molecules measured by optical trapping nanometry. AB - We have analyzed the mechanics of individual kinesin molecules by optical trapping nanometry. A kinesin molecule was adsorbed onto a latex bead, which was captured by an optical trap and brought into contact with an axoneme that was bound to a glass surface. The displacement of kinesin during force generation was determined by measuring the position of the beads with nanometer accuracy. As the displacement of kinesin was attenuated because of the compliance of the kinesin to-bead and kinesin-to-microtubule linkages, the compliance was monitored during force generation and was used to correct the displacement of kinesin. Thus the velocity and the unitary steps could be obtained accurately over a wide force range. The force-velocity curves were linear from 0 to a maximum force at 10 microM and 1 mM ATP, and the maximum force was approximately 7 pN, which is larger by approximately 30% than values previously reported. Kinesin exhibited forward and occasionally backward stepwise displacements with a size of approximately 8 nm. The histograms of step dwell time show a monotonic decrease with time. Model calculations indicate that each kinesin head steps by 16-nm, whereas kinesin molecule steps by 8-nm. PMID- 9336197 TI - Spectroscopic study of conformational changes in subdomain 1 of G-actin: influence of divalent cations. AB - Temperature dependence of the fluorescence intensity and anisotropy decay of N (iodoacetyl)-N'-(5-sulfo-1-naphthyl)ethylenediamine attached to Cys374 of actin monomer was investigated to characterize conformational differences between Ca- and Mg-G-actin. The fluorescence lifetime is longer in Mg-G-actin than that in Ca G-actin in the temperature range of 5-34 degrees C. The width of the lifetime distribution is smaller by 30% in Mg-saturated actin monomer at 5 degrees C, and the difference becomes negligible above 30 degrees C. The semiangle of the cone within which the fluorophore can rotate is larger in Ca-G-actin at all temperatures. Electron paramagnetic resonance measurements on maleimide spin labeled (on Cys374) monomer actin gave evidence that exchange of Ca2+ for Mg2+ induced a rapid decrease in the mobility of the label immediately after the addition of Mg2+. These results suggest that the C-terminal region of the monomer becomes more rigid as a result of the replacement of Ca2+ by Mg2+. The change can be related to the difference between the polymerization abilities of the two forms of G-actin. PMID- 9336198 TI - Measurement of nucleotide release kinetics in single skeletal muscle myofibrils during isometric and isovelocity contractions using fluorescence microscopy. AB - Rabbit psoas muscle myofibrils, in the presence of the fluorescent nucleotide analog 2'(3')-O-[N-[2-[[Cy3]amido]ethyl]carbamoyl]-adenosine 5' triphosphate (Cy3 EDA-ATP), showed selective fluorescence staining of the A-band with a reduced fluorescence at the M-line. Addition of Cy3-EDA-ATP to a myofibril in the presence of Ca2+ caused auxotonic shortening against a compliant glass microneedle. These results indicate that Cy3-EDA-ATP is a substrate for myosin in the myofibril system. The kinetics of nucleotide release from a single myofibril, held isometrically between two needles, were measured by the displacement of prebound Cy3-EDA-ATP on flash photolysis of caged ATP. The A-band fluorescence of the myofibril decayed exponentially with a rate constant of 0.3 s(-1) at 8 degrees C, an order of magnitude faster than that for isolated thick filaments in the absence of actin. When a myofibril was imposed to shorten with a constant velocity by a piezoelectric actuator, the nucleotide displacement rate constant initially increased to 0.7 s(-1) with increasing shortening velocity and then declined with a further increase in shortening velocity. These results demonstrate that the displacement rates of Cy3-EDA-nucleotides bound to the cross bridges in the contracting myofibril reflect a process that shows strain dependence. PMID- 9336199 TI - Titin elasticity and mechanism of passive force development in rat cardiac myocytes probed by thin-filament extraction. AB - Titin (also known as connectin) is a giant filamentous protein whose elastic properties greatly contribute to the passive force in muscle. In the sarcomere, the elastic I-band segment of titin may interact with the thin filaments, possibly affecting the molecule's elastic behavior. Indeed, several studies have indicated that interactions between titin and actin occur in vitro and may occur in the sarcomere as well. To explore the properties of titin alone, one must first eliminate the modulating effect of the thin filaments by selectively removing them. In the present work, thin filaments were selectively removed from the cardiac myocyte by using a gelsolin fragment. Partial extraction left behind approximately 100-nm-long thin filaments protruding from the Z-line, whereas the rest of the I-band became devoid of thin filaments, exposing titin. By applying a much more extensive gelsolin treatment, we also removed the remaining short thin filaments near the Z-line. After extraction, the extensibility of titin was studied by using immunoelectron microscopy, and the passive force-sarcomere length relation was determined by using mechanical techniques. Titin's regional extensibility was not detectably affected by partial thin-filament extraction. Passive force, on the other hand, was reduced at sarcomere lengths longer than approximately 2.1 microm, with a 33 +/- 9% reduction at 2.6 microm. After a complete extraction, the slack sarcomere length was reduced to approximately 1.7 microm. The segment of titin near the Z-line, which is otherwise inextensible, collapsed toward the Z-line in sarcomeres shorter than approximately 2.0 microm, but it was extended in sarcomeres longer than approximately 2.3 microm. Passive force became elevated at sarcomere lengths between approximately 1.7 and approximately 2.1 microm, but was reduced at sarcomere lengths of >2.3 microm. These changes can be accounted for by modeling titin as two wormlike chains in series, one of which increases its contour length by recruitment of the titin segment near the Z-line into the elastic pool. PMID- 9336200 TI - Guanine tetraplex formation by short DNA fragments containing runs of guanine and cytosine. AB - Using CD spectroscopy, guanine tetraplex formation was studied with short DNA fragments in which cytosine residues were systematically added to runs of guanine either at the 5' or 3' ends. Potassium cations induced the G-tetraplex more easily with fragments having the guanine run at the 5' end, which is just an opposite tendency to what was reported for (G+T) oligonucleotides. However, the present (G+C) fragments simultaneously adopted other conformers that complicated the analysis. We demonstrate that repeated freezing/thawing, performed at low ionic strength, is a suitable method to exclusively stabilize the tetraplex in the (G+C) DNA fragments. In contrast to KCl, the repeated freeze/thaw cycles better stabilized the tetraplex with fragments having the guanine run on the 3' end. The tendency of guanine blocks to generate the tetraplex destabilized the d(G5).d(C5) duplex whose strands dissociated, giving rise to a stable tetraplex of (dG5) and single-stranded (dC5). In contrast to d(G3C3) and d(G5C5), repeated freezing/thawing induced the tetraplex even with the self-complementary d(C3G3) or d(C5G5); hence the latter oligonucleotides preferred the tetraplex to the apparently very stable duplex. The tetraplexes only included guanine blocks while the 5' end cytosines interfered neither with the tetraplex formation nor the tetraplex structure. PMID- 9336201 TI - pH-dependent specific binding and combing of DNA. AB - Recent developments in the rapid sequencing, mapping, and analysis of DNA rely on the specific binding of DNA to specially treated surfaces. We show here that specific binding of DNA via its unmodified extremities can be achieved on a great variety of surfaces by a judicious choice of the pH. On hydrophobic surfaces the best binding efficiency is reached at a pH of approximately 5.5. At that pH a approximately 40-kbp DNA is 10 times more likely to bind by an extremity than by a midsegment. A model is proposed to account for the differential adsorption of the molecule extremities and midsection as a function of pH. The pH-dependent specific binding can be used to align anchored DNA molecules by a receding meniscus, a process called molecular combing. The resulting properties of the combed molecules will be discussed. PMID- 9336203 TI - Evidence for a controlling role of water in producing the native bacteriorhodopsin structure. AB - The experiments reported in this paper, based on reconstitution of bacteriorhodopsin (bR) from apomembrane at varying environmental conditions, demonstrate that the presence of water is a controlling factor in generating a native wild-type bR conformation. If water is lacking during this reconstitution process, then a non-native bR structure is formed that exhibits altered M formation and decay kinetics, as well as different behavior following extensive dehydration. It is shown that mutants affecting the ability of bR to form appropriate structures of water in specific protein cavities also affect the ability to generate a native bR conformation. The results suggest that aspartic acid 96 plays a major role in anchoring the appropriate water structure conformation associated with bR. It is also demonstrated that the glutamic acid 204 residue is pivotal in controlling the protein/water affinity. This water affinity can be further controlled by modifying the charge environment of the protein with altered pH. These data, based on kinetic absorption spectroscopy and Fourier transform infrared spectroscopy, highlight the central role of water in this protein. PMID- 9336202 TI - Fourier transform infrared double-flash experiments resolve bacteriorhodopsin's M1 to M2 transition. AB - The orientation of the central proton-binding site, the protonated Schiff base, away from the proton release side to the proton uptake side is crucial for the directionality of the proton pump bacteriorhodopsin. It has been proposed that this movement, called the reprotonation switch, takes place in the M1 to M2 transition. To resolve the molecular events in this M1 to M2 transition, we performed double-flash experiments. In these experiments a first pulse initiates the photocycle and a second pulse selectively drives bR molecules in the M intermediate back into the BR ground state. For short delay times between initiating and resetting pulses, most of the M molecules being reset are in the M1 intermediate, and for longer delay times most of the reset M molecules are in the M2 intermediate. The BR-M1 and BR-M2 difference spectra are monitored with nanosecond step-scan Fourier transform infrared spectroscopy. Because the Schiff base reprotonation rate is kM1 = 0.8 x 10(7) s(-1) in the light-induced M1 back reaction and kM2 = 0.36 x 10(7) s(-1) in the M2 back-reaction, the two different M intermediates represent two different proton accessibility configurations of the Schiff base. The results show only a minute movement of one or two peptide bonds in the M1 to M2 transition that changes the interaction of the Schiff base with Y185. This backbone movement is distinct from the larger one in the subsequent M to N transition. No evidence of a chromophore isomerization is seen in the M1 to M2 transition. Furthermore, the results show time-resolved reprotonation of the Schiff base from D85 in the M photo-back-reaction, instead of from D96, as in the conventional cycle. PMID- 9336204 TI - Pump-probe anisotropies of Fenna-Matthews-Olson protein trimers from Chlorobium tepidum: a diagnostic for exciton localization? AB - Exciton calculations on symmetric and asymmetric Fenna-Matthews-Olson (FMO) trimers, combined with absorption difference anisotropy measurements on FMO trimers from the green bacterium Chlorobium tepidum, suggest that real samples exhibit sufficient diagonal energy disorder so that their laser-excited exciton states are noticeably localized. Our observed anisotropies are clearly inconsistent with 21-pigment exciton simulations based on a threefold-symmetric FMO protein. They are more consistent with a 7-pigment model that assumes that the laser-prepared states are localized within a subunit of the trimer. Differential diagonal energy shifts of 50 cm(-1) between symmetry-related pigments in different subunits are large enough to cause sharp localization in the stationary states; these shifts are commensurate with the approximately 95 cm(-1) inhomogeneous linewidth of the lowest exciton levels. Experimental anisotropies (and by implication steady-state linear and circular dichroism) likely arise from statistical averaging over states with widely contrasting values of these observables, in consequence of their sensitivity to diagonal energy disorder. PMID- 9336205 TI - Studies of cation binding in ZnCl2-regenerated bacteriorhodopsin by x-ray absorption fine structures: effects of removing water molecules and adding Cl- ions. AB - The binding of Zn2+ in Zn2+-regenerated bacteriorhodopsin (bR) was studied under various conditions by x-ray absorption fine structures (XAFS). The 0.9:1 and 2:1 Zn2+:bR samples gave similar XAFS spectra, suggesting that Zn2+ might have only one strong binding site in bR. It was found that in aqueous bR solution, Zn2+ has an average of six oxygen or nitrogen ligands. Upon drying, two ligands are lost, suggesting the existence of two weakly bound water ligands near the cation binding site in bacteriorhodopsin. When excess Cl- ions were present before drying in the Zn2+-regenerated bR samples, it was found that two of the ligands were replaced by Cl- ions in the dried film, whereas two remain unchanged. The above observations suggest that Zn2+ has three types of ligands in regenerated bR (referred to as types I, II, and III). Type I ligands are strongly bound. These ligands cannot be removed by drying or by exchanging with Cl- ions. Type II ligands cannot be removed by drying, but can be replaced by Cl- ligands. Type III ligands are weakly bound to the metal cation and are most likely water molecules that can be removed by evaporation under vacuum or by drying with anhydrous CaSO4. The results are discussed in terms of the possible structure of the strongly binding site of Zn2+ in bR. PMID- 9336206 TI - Differences in hydration structure near hydrophobic and hydrophilic amino acids. AB - We use molecular dynamics to simulate recent neutron scattering experiments on aqueous solutions of N-acetyl-leucine-amide and N-acetyl-glutamine-amide, and break down the total scattering function into contributions from solute-solute, solute-water, water-water, and intramolecular correlations. We show that the shift of the main diffraction peak to smaller angle that is observed for leucine, but not for glutamine, is attributable primarily to alterations in water-water correlations relative to bulk. The perturbation of the water hydrogen-bonded network extends roughly two solvation layers from the hydrophobic side chain surface, and is characterized by a distribution of hydrogen bonded ring sizes that are more planar and are dominated by pentagons in particular than those near the hydrophilic side chain. The different structural organization of water near the hydrophobic solute that gives rise to the inward shift in the main neutron diffraction peak under ambient conditions may also provide insight into the same directional shift for pure liquid water as it is cooled and supercooled. PMID- 9336207 TI - Involvement of water molecules in the association of monoclonal antibody HyHEL-5 with bobwhite quail lysozyme. AB - Fluorescence polarization spectroscopy and isothermal titration calorimetry were used to study the influence of osmolytes on the association of the anti-hen egg lysozyme (HEL) monoclonal antibody HyHEL-5 with bobwhite quail lysozyme (BWQL). BWQL is an avian species variant with an Arg-->Lys mutation in the HyHEL-5 epitope, as well as three other mutations outside the HyHEL-5 structural epitope. This mutation decreases the equilibrium association constant of HyHEL-5 for BWQL by over 1000-fold as compared to HEL. The three-dimensional structure of this complex has been obtained recently. Fluorescein-labeled BWQL, obtained by labeling at pH 7.5 and purified by hydrophobic interaction chromatograpy, bound HyHEL-5 with an equilibrium association constant close to that determined for unlabeled BWQL by isothermal titration calorimetry. Fluorescence titration, stopped-flow kinetics, and isothermal titration calorimetry experiments using various concentrations of the osmolytes glycerol, ethylene glycol, and betaine to perturb binding gave a lower limit of the uptake of approximately 6-12 water molecules upon formation of the HyHEL-5/BWQL complex. PMID- 9336209 TI - A molecular dynamics study of Fe2S2 putidaredoxin: multiple conformations of the C-terminal region. AB - Putidaredoxin (Pdx) plays an essential role as an electron donor and effector in the biochemical cycle involving cytochrome P450cam. Only recently has an NMR derived structure for this protein been published, but because of the presence of a paramagnetic Fe2S2 center, the NMR assignment could not be completed for residues within a region of 8 A around the active site. That region was modeled by homology with a related protein. The structural refinement for those experiments was done in vacuum, without the use of electrostatic terms in the force field. The present manuscript will describe and discuss a series of long time, unrestrained, solution molecular dynamic runs for this system. Results will be presented that construct a molecular-level picture that rationalizes experimental results concerning the conformation and mobility of the C-terminal residue Trp106. At least two different conformers are found for this residue during the simulations. The time scale for interconversion between them is found to be in the subnanosecond regime. The results presented here open the possibility for studying binding and electron transfer between Pdx and P450cam, in a framework that allows for dynamical information to be used during the computational process, instead of the single structures deposited on the protein data base. PMID- 9336208 TI - Picosecond molecular motions in bacteriorhodopsin from neutron scattering. AB - The characteristics of internal molecular motions of bacteriorhodopsin in the purple membrane have been studied by quasielastic incoherent neutron scattering. Because of the quasihomogeneous distribution of hydrogen atoms in biological molecules, this technique enables one to study a wide variety of intramolecular motions, especially those occurring in the picosecond to nanosecond time scale. We performed measurements at different energy resolutions with samples at various hydration levels within a temperature range of 10-300 K. The analysis of the data revealed a dynamical transition at temperatures Td between 180 K and 220 K for all motions resolved at time scales ranging from 0.1 to a few hundred picoseconds. Whereas below Td the motions are purely vibrational, they are predominantly diffusive above Td, characterized by an enormously broad distribution of correlation times. The variation of the hydration level, on the other hand, mainly affects motions slower than a few picoseconds. PMID- 9336211 TI - No salting-in of lysozyme chloride observed at low ionic strength over a large range of pH. AB - Solubility of lysozyme chloride was determined in the absence of added salt and in the presence of 0.05-1.2 M NaCl, starting from isoionic lysozyme, which was then brought to pH values from 9 to 3 by addition of HCl. The main observation is the absence of a salting-in region whatever the pH studied. This is explained by a predominant electrostatic screening of the positively charged protein and/or by adsorption of chloride ions by the protein. The solubility increases with the protein net charge at low ionic strength, but the reverse is observed at high ionic strength. The solubility of lysozyme chloride seems to become independent of ionic strength at pH approximately 9.5, which is interpreted as a shift of the isoionic pH (10.8) to an isoelectric pH due to chloride binding. The crystallization at very low ionic strength, where lysozyme crystallizes at supersaturation values as low as 1.1, amplifies the effect of pH on protein solubility. Understanding the effect of the net charge and of ionic strength on protein-protein interactions is valuable not only for protein crystal growth but more generally also for the formation of protein-protein or protein-ligand complexes. PMID- 9336210 TI - Lys515-Lys492 cross-linking by DIDS interferes with substrate utilization by the sarcoplasmic reticulum ATPase. AB - Sarcoplasmic reticulum (SR) Ca2+ ATPase was derivatized with 4,4' diisothiocyanatostilbene-2,2'-sulfonic acid (DIDS), and complete enzyme inactivation was produced with a molecular stoichiometry of one DIDS per ATPase. It was determined by peptide analysis and sequencing that Lys492 and Lys515 were the ATPase residues derivatized by DIDS. Lack of electrophoretic resolution of the two peptide fragments that result from a single tryptic cut at Arg505 demonstrated that the two derivatized residues were cross-linked. Cross-linking of Lys492 and Lys515 by DIDS interfered with ATPase utilization of both ATP and p nitrophenyl phosphate substrates, whereas derivatization of only Lys515 with fluorescein isothiocyanate interfered with ATPase utilization of ATP but not of p nitrophenyl phosphate. Cross-linking with DIDS implies a distance of approximately 13 A between Lys492 and Lys515, which corresponds to the length of ATP bound in an extended configuration. Therefore, within the groove of the nucleotide binding domain, the ATP substrate is positioned with the adenosine moiety near Lys515 and its terminal phosphate near Lys492. PMID- 9336212 TI - Self-assembly of collagen fibers. Influence of fibrillar alignment and decorin on mechanical properties. AB - Collagen is the primary structural element in extracellular matrices. In the form of fibers it acts to transmit forces, dissipate energy, and prevent premature mechanical failure in normal tissues. Deformation of collagen fibers involves molecular stretching and slippage, fibrillar slippage, and, ultimately, defibrillation. Our laboratory has developed a process for self-assembly of macroscopic collagen fibers that have structures and mechanical properties similar to rat tail tendon fibers. The purpose of this study is to determine the effects of subfibrillar orientation and decorin incorporation on the mechanical properties of collagen fibers. Self-assembled collagen fibers were stretched 0 50% before cross-linking and then characterized by microscopy and mechanical testing. Results of these studies indicate that fibrillar orientation, packing, and ultimate tensile strength can be increased by stretching. In addition, it is shown that decorin incorporation increases ultimate tensile strength of uncross linked fibers. Based on the observed results it is hypothesized that decorin facilitates fibrillar slippage during deformation and thereby improves the tensile properties of collagen fibers. PMID- 9336213 TI - Driving proteins off DNA using applied tension. AB - Proteins that bind DNA so as to reduce its end-to-end length can be dissociated by application of force. The thermodynamics of this process are discussed, with special attention to the case of histones bound to DNA (i.e., a string of nucleosomes, or chromatin fiber). The histone octamer is predicted to be driven off chromatin fiber for tensions >2 piconewtons. PMID- 9336214 TI - Entropic drive in the sarcoplasmic reticulum ATPase interaction with Mg2+ and Pi. AB - Thermodynamic quantities for the binding of Mg2+ (in the presence of Ca2+) and Pi (in the presence of Mg2+ and absence of Ca2+) to sarcoplasmic reticulum ATPase were determined from isothermal titration calorimetry measurements at 25 degrees C. Mg2+ and Pi are involved in reversal of the ATPase hydrolytic reaction, and their interactions with the ATPase are conveniently studied under equilibrium conditions. We found that the Mg2+ binding reaction is endothermic with a binding constant (Kb) = 142 +/- 4 M(-1), a binding enthalpy of 180 +/- 3 kJ mol(-1), and an entropy contribution (TdeltaSb) = 192 +/- 3 kJ mol(-1). Similarly, Pi binding is also an endothermic reaction with Kb = 167 +/- 17 M(-1), deltaHb = 65.3 +/- 5.4 kJ mol(-1), and TdeltaSb = 77.9 +/- 5.4 kJ mol(-1). Our measurements demonstrate that the ATPase can absorb heat from the environment upon ligand binding, and emphasize the important role of entropic mechanisms in energy transduction by this enzyme. PMID- 9336215 TI - AFM review study on pox viruses and living cells. AB - Single living cells were studied in growth medium by atomic force microscopy at a high--down to one image frame per second--imaging rate over time periods of many hours, stably producing hundreds of consecutive scans with a lateral resolution of approximately 30-40 nm. The cell was held by a micropipette mounted onto the scanner-piezo as shown in Haberle, W., J. K. H. Horber, and G. Binnig. 1991. Force microscopy on living cells. J. Vac. Sci. Technol. B9:1210-0000. To initiate specific processes on the cell surface the cells had been infected with pox viruses as reported earlier and, most likely, the liberation of a progeny virion by the still-living cell was observed, hence confirming and supporting earlier results (Haberle, W., J. K. H. Horber, F. Ohnesorge, D. P. E. Smith, and G. Binnig. 1992. In situ investigations of single living cells infected by viruses. Ultramicroscopy. 42-44:1161-0000; Horber, J. K. H., W. Haberle, F. Ohnesorge, G. Binnig, H. G. Liebich, C. P. Czerny, H. Mahnel, and A. Mayr. 1992. Investigation of living cells in the nanometer regime with the atomic force microscope. Scanning Microscopy. 6:919-930). Furthermore, the pox viruses used were characterized separately by AFM in an aqueous environment down to the molecular level. Quasi-ordered structural details were resolved on a scale of a few nm where, however, image distortions and artifacts due to multiple tip effects are probably involved--just as in very high resolution (<15-20 nm) images on the cells. Although in a very preliminary manner, initial studies on the mechanical resonance properties of a single living (noninfected) cell, held by the micropipette, have been performed. In particular, frequency response spectra were recorded that indicate elastic properties and enough stiffness of these cells to make the demonstrated rapid scanning of the imaging tip plausible. Measurements of this kind, especially if they can be proven to be cell-type specific, may perhaps have a large potential for biomedical applications. Images of these living cells were also recorded in the widely known (e.g., Radmacher, M., R. W. Tillmann, and H. E. Gaub. 1993. Imaging viscoelasticity by force modulation with the atomic force microscope. Biophys. J. 64:735-742) force modulation mode, yet at one low modulation frequency of approximately 2 kHz. (Note: After the cells were attached to the pipette by suction, they first deformed significantly and then reassumed their original spherical shape, which they also acquire when freely suspended in solution, to a great extent with the exception of the portion adjusting to the pipette edge geometry after approximately 0.5-1 h, which occurred in almost the same manner with uninfected cells, and those that had been infected several hours earlier. This seems to be a process which is at least actively supported by the cellular cytoskeleton, rather than a mere osmotic pressure effect induced by electrolyte transport through the membrane. Furthermore, several hours postinfection (p.i.) infected cells developed many optically visible refraction effects, which appeared as small dark spots in the light microscope, that we believed to be the regions in the cell plasma where viruses are assembled; this is known from the literature on electron microscopy on pox-infected cells and referred to there as "virus factories" (e.g., Moss, B. 1986. Replication of pox viruses. In Fundamental Virology, B. N. Fields and D. M. Knape, editors. Raven Press, New York. 637-655). Therefore, we assume that the cells stay alive during imaging, in our experience for approximately 30-45 h p.i.). PMID- 9336216 TI - Site-specific interaction of thrombin and inhibitors observed by fluorescence correlation spectroscopy. AB - We report on the application of fluorescence correlation spectroscopy (FCS) to observe the interaction between thrombin and thrombin inhibitors. Two site specific fluorescent labels were used to distinguish between inhibitors directed to the active site, the exosite, or both binding sites of thrombin. For several well-known inhibitors of thrombin, the binding sites observed by FCS correspond to previous studies. The interaction of the recently discovered thrombin inhibitor ornithodorin from the tick Ornithodorus moubata with thrombin was investigated. It was found that this inhibitor, like hirudin and rhodniin, binds to both the active site and exosite of thrombin simultaneously. This study shows the feasibility of FCS as a sensitive and selective method for observing protein ligand interactions. As an additional technique, simultaneous labeling with both fluorescent labels was successfully demonstrated. PMID- 9336217 TI - Lipid-gramicidin interactions using two-dimensional Fourier-transform electron spin resonance. AB - The application of two-dimensional Fourier-transform electron-spin-resonance (2D FT-ESR) to the study of lipid/gramicidin A (GA) interactions is reported. It is shown that 2D-FT-ESR spectra provide substantially enhanced spectral resolution to changes in the dynamics and ordering of the bulk lipids (as compared with cw ESR spectra), that result from addition of GA to membrane vesicles of dipalmitoylphosphatidylcholine (DPPC) in excess water containing 16-PC as the lipid spin label. The agreement between the theory of Lee, Budil, and Freed and experimental results is very good in the liquid crystalline phase. Both the rotational and translational diffusion rates of the bulk lipid are substantially decreased by addition of GA, whereas the ordering is only slightly increased, for a 1:5 ratio of GA to lipid. The slowing effect on the diffusive rates of adding GA in the gel phase is less pronounced. It is suggested that the spectral fits in this phase would be improved with a more detailed dynamic model. No significant evidence is found in the 2D-FT-ESR spectra for a second immobilized component upon addition of GA, which is in contrast to cw-ESR. It is shown from simulations of the observed 2D-FT-ESR spectra that the additional component seen in cw-ESR spectra, and usually attributed to "immobilized" lipid, is inconsistent with its being characterized by increased ordering, according to a model proposed by Ge and Freed, but it would be consistent with the more conventional model of a significantly reduced diffusional rate. This is because the 2D-FT-ESR spectra exhibit a selectivity, favoring components with longer homogeneous relaxation times, T2. The homogeneous linewidths of the 2D-FT-ESR autopeaks appear to broaden as a function of mixing time. This apparent broadening is very likely due to the process of cooperative order director fluctuations (ODF) of the lipids in the vesicle. This real-time observation of ODF is distinct from, but appears in reasonable agreement with, NMR results. It is found that addition of GA to give the 1:5 ratio has only a small effect on the ODF, but there is a significant temperature dependence. PMID- 9336218 TI - Laurdan solvatochromism: solvent dielectric relaxation and intramolecular excited state reaction. AB - Absorption, steady-state, and time-resolved fluorescence measurements have been performed on laurdan dissolved either in white viscous apolar solvents or in ethanol as a function of temperature. The heterogeneity of the absorption spectra in white oils or in ethanol is consistent with semiempirical calculations performed previously on Prodan. From steady-state and time-resolved fluorescence measurements in apolar media, an excited state reaction is evidenced. The bimodal lifetime distribution determined from the maximum entropy method (MEM) analysis is attributed to the radiative deexcitation of a "locally excited" (LE) state and of a "charge transfer" (CT) state, whereas a very short component (20 ps), the sign and the amplitude of which depend on the emission wavelength, is attributed to the kinetics of the interconvertion reaction. The observation of an isoemissive point in the temperature range from -50 degrees C to -110 degrees C in ethanol suggests an interconvertion between two average excited-state populations: unrelaxed and solvent-relaxed CT states. A further decrease in temperature (-190 degrees C), leading to frozen ethanol, induces an additional and important blue shift. This low temperature spectrum is partly attributed to the radiative deexcitation of the LE state. Time-resolved emission spectra (TRES) measurements at -80 degrees C in the ethanol liquid phase show a large spectral shift of approximately 2500 cm(-1) (stabilization energy of the excited state: 7.1 kcal x M(-1)). The time-dependent fluorescence shift (TDFS) is described for its major part by a nanosecond time constant. The initial part of the spectral shift reveals, however, a subnanosecond process that can be due to fast internal solvent reorientation and/or to intramolecular excited-state reactions. These two relaxation times are also detected in the analysis of the fluorescence decays in the middle range of emission energy. The activation energy of the longest process is approximately 3 kcal x M(-1). At -190 degrees C, one subnanosecond and one nanosecond excited-state reactions are also evidenced. They are likely due to intramolecular rearrangements after the excitation, leading to the CT state and not to solvent relaxation, which is severely hindered in these temperature conditions. Therefore, both intramolecular and solvent relaxations are responsible for the large Stokes shift displayed by this probe as a function of solvent polarity. A possible scheme is proposed for the deexcitation pathway, taking into account the kinetics observed in these different solvents. PMID- 9336219 TI - Vibrational spectra of individual millimeter-size membrane patches using miniature infrared waveguides. AB - We have used miniature planar IR waveguides, consisting of Ge strips 30-50 microm thick and 2 mm wide, as evanescent-wave sensors to detect the mid-(IR) evanescent wave absorbance spectra of small areas of biomolecular monolayers and multilayers. Examples include picomolar quantities of an integral transmembrane protein (bacteriorhodopsin) and lipid (dimyristoyl phosphatidylcholine). IR bands due to the protein and lipid components of the plasma membrane of individual 1.5 mm-diameter devitellinized Xenopus laevis oocytes, submerged in buffer and sticking to the waveguide surface, were also detected. A significant improvement in sensitivity was observed, as compared to previous sizes and geometries of evanescent-wave sensors (e.g., commercially available internal reflection elements or tapered optical fibers). These measurements suggest the feasibility of using such miniature supported planar IR waveguides to observe structural changes in transmembrane proteins functioning in vivo in single cells. PMID- 9336221 TI - Regional differences in synaptogenesis in human cerebral cortex. AB - The formation of synaptic contacts in human cerebral cortex was compared in two cortical regions: auditory cortex (Heschl's gyrus) and prefrontal cortex (middle frontal gyrus). Synapse formation in both cortical regions begins in the fetus, before conceptual age 27 weeks. Synaptic density increases more rapidly in auditory cortex, where the maximum is reached near postnatal age 3 months. Maximum synaptic density in middle frontal gyrus is not reached until after age 15 months. Synaptogenesis occurs concurrently with dendritic and axonal growth and with myelination of the subcortical white matter. A phase of net synapse elimination occurs late in childhood, earlier in auditory cortex, where it has ended by age 12 years, than in prefrontal cortex, where it extends to midadolescence. Synaptogenesis and synapse elimination in humans appear to be heterochronous in different cortical regions and, in that respect, appears to differ from the rhesus monkey, where they are concurrent. In other respects, including overproduction of synaptic contacts in infancy, persistence of high levels of synaptic density to late childhood or adolescence, the absolute values of maximum and adult synaptic density, and layer specific differences, findings in the human resemble those in rhesus monkeys. PMID- 9336220 TI - Mechanics of microtubule bundles in pillar cells from the inner ear. AB - The mechanical properties of cross-linked microtubule bundles were measured from outer pillar cells isolated from the mammalian inner ear. Measurements were made using a three-point bending test and were incorporated into a mathematical model designed to distinguish between the stiffness contributions from microtubules and their cross-linking proteins. Outer pillar cells were composed of 1000-3000 parallel bundled microtubules in a square array that was interdigitated and cross linked with actin filaments. The average midpoint bending stiffness of intact cells was 7 x 10(-4) N/m. After removal of both the actin filaments and cross links with detergent in the presence of DNase I, the square array was disrupted and the stiffness decreased by a factor of 4, to 1.7 x 10(-4) N/m. The bending modulus for individual microtubules was calculated to be 7 x 10(-23) Nm2, and the Young's modulus for these 15 protofilament microtubules was 2 x 10(9) Pa. The shear modulus between microtubules in intact cells was calculated to be 10(3) Pa. It was concluded that cross-linking proteins provided shear resistance between microtubules, which resulted in a fourfold increase in stiffness. The model can be used to estimate the mechanical properties of cross-linked microtubule bundles in cells from which direct measurements are not available. PMID- 9336222 TI - Histogenesis of ferret somatosensory cortex. AB - The ferret has emerged as an important animal model for the study of neocortical development. Although detailed studies of the birthdates of neurons populating the ferret visual cortex are available, the birthdates of neurons that reside in somatosensory cortex have not been determined. The current study used bromodeoxyuridine to establish when neurons inhabiting the somatosensory cortex are generated in the ferret; some animals also received injections of [3H]thymidine. In contrast to reports of neurogenesis in ferret visual cortex, most neurons populating the somatosensory cortex have been generated by birth. Although components of all somatosensory cortical layers have been produced at postnatal day 0, the layers are not distinctly formed but develop over a period of several weeks. A small number of neurons continue to be produced for a few days postnatally. The majority of cells belonging to a given layer are born over a period of approximately 3 days, although the subplate and last (layer 2) generated layer take somewhat longer. Although neurogenesis of the neocortex begins along a similar time line for visual and somatosensory cortex, the neurons populating the visual cortex lag substantially during the generation of layer 4, which takes more than 1 week for ferret visual cortex. Layer formation in ferret somatosensory cortex follows many established principles of cortical neurogenesis, such as the well-known inside-out development of cortical layers and the rostro-to-caudal progression of cell birth. In comparison with the development of ferret visual cortex, however, the generation of the somatosensory cortex occurs remarkably early and may reflect distinct differences in mechanisms of development between the two sensory areas. PMID- 9336223 TI - Timecourse of development of the wallaby trigeminal pathway: III. Thalamocortical and corticothalamic projections. AB - The development of trigeminal projections between the thalamus and cortex has been investigated in the marsupial mammal, the wallaby, by using a carbocyanine dye, horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP), Neurobiotin, and biocytin as pathway tracers. The appearance of whisker-related patterns in the cortex in relation to their appearance in the brainstem and thalamus was examined, as was the presence or absence of a waiting period for thalamocortical afferents and the identity of the first cortical cells to project to the thalamus. Thalamic afferents first reached the cortex at postnatal day (P) 15 and were distributed up to the deep edge of the compact cell zone in the superficial cortical plate throughout development, in both dye and WGA-HRP labelled material, with no evidence of a waiting period. The initial corticothalamic projection, detected by retrograde transport of WGA-HRP from the thalamus, occurred at P60 from layer 5 cells. This was confirmed by labelling of corticothalamic axons after cortical injections of Neurobiotin and biocytin. Scattered, labelled cells seen before P60 after dye labelling from the thalamus presumably resulted from transcellular labelling via thalamic afferents. Clustering of afferents in layer 4 and cell bodies and their dendrites in layers 5 and 6 first occurred simultaneously at P76. There is a sequential onset of pattern formation, first in brainstem, then in thalamus, and finally in cortex, with a long delay between afferent arrival and pattern formation at each level. Independent regulation at each level, likely depending on target maturation, is suggested. PMID- 9336224 TI - Somatosensory fovea in the star-nosed mole: behavioral use of the star in relation to innervation patterns and cortical representation. AB - Eleven fleshy appendages, or rays, surround each of the nostrils of the star nosed mole. Each ray is covered with tactile sensory organs, and each ray is represented in the cortex by a stripe of tissue visible in brain sections processed for cytochrome oxidase. Here we report that the 11th, ventral ray is the behavioral focus of the nose. This ray is preferentially used to explore prey items by touch, in a behavior pattern similar to the use of a fovea in the visual system. After prey is first contacted with any nasal ray, subsequent touches are centered on the 11th ray. Although the 11th ray is small and has relatively few sensory organs on its surface, it has the largest cortical representation, greatest area of cortex per sensory organ, and the highest innervation density per sensory organ. In addition, the average area of cortex per primary afferent is highest for this ray. We refer to the differential magnification of first order afferents in the cortical representation as "afferent magnification." The patterns of both cortical magnification (cortical area per sensory organ) and afferent magnification (cortical area per afferent) of the rays correlated highly with the distribution of touches across the nose scored from videotaped behavior. A simple model of star-nosed mole behavior predicts the distribution of touches across the rays and also correlates highly with both the actual pattern of behavior and the patterns of cortical magnification observed. PMID- 9336225 TI - Delayed hypoglossal-facial nerve suture after predegeneration of the peripheral facial nerve stump improves the innervation of mimetic musculature by hypoglossal motoneurons. AB - Surgical reconstruction of the facial nerve is common clinical practice following destruction of the intracranial facial nerve. Delayed hypoglossal-facial anastomosis (HFA) is the procedure of choice, although the effect of delay on outcome remains unclear. To study the effect of delayed anastomosis on reinnervation, we sutured the proximal stump of a freshly transected hypoglossal nerve of Wistar rats to the distal stump of the ipsilateral facial nerve, which had been transected 7-56 days earlier. Animals that had received HFA without delay served as the control group. Forty days after HFA, horseradish peroxidase (HRP) was injected into the whisker pad; 2 days later, the animals were killed. Reinnervation was assessed by determining the proportion of labeled neuronal cell bodies in the brainstem. The control group had 68% reinnervation of these muscles by hypoglossal neurons and had 32% reinnervation by facial neurons. When the distal facial nerve had been allowed to degenerate for 7 days before HFA, reinnervation of the hypoglossal nerve decreased to 54%, and reinnervation by the facial nerve increased to 46%. However, after a delay of 10-56 days, the hypoglossal fraction increased and stabilized at 77%, and the facial motoneuron fraction decreased to 23%. The presence of new neuromuscular junctions was confirmed by HRP labeling of motor end plates in vivo and by electromyography. We conclude that, under the conditions of hypoglossal-facial crossed nerve suture, the predegeneration of the distal stump of a transected facial nerve enhances the reinnervation of facial muscles by hypoglossal axonal sprouts. PMID- 9336226 TI - Melatonin in a primitive metazoan: seasonal changes of levels and immunohistochemical visualization in neurons. AB - Monthly day/night melatonin activity profiles were determined by radioimmunoassay over a 13-month period in the colonial anthozoan Renilla kollikeri, and no daily rhythmic oscillation was found. Averaging those monthly values yielded a seasonal quantitative rhythm in both colonial and non-colonial tissues of this cnidarian, with spring and summer levels found to be four to five times higher than autumn and winter ones. The annual rise, which occurred in two successive Aprils, coincided with the first stages of sexual maturation in R. kollikeri. Immunohistochemistry with a melatonin antibody raised in sheep revealed an exclusively neuronal distribution of melatonin-immunoreactivity (MEL-IR) in the endodermal septal filaments wrapped around gametophores, in endodermal walls of the rachis, and in the ectoderm of polyps. The MEL-IR ectodermal neurons shared many morphological features with serotonin-immunoreactive (5-HT-IR) neurons previously described in this animal but showed either weak or absent 5-HT-IR in double-labelling experiments. In contrast, MEL-IR and 5-HT-IR were strongly colocalized in endodermal neurons. These results indicate that melatonin is not a daily photoperiodic messenger but may instead act as a seasonal marker for reproduction in this cnidarian. We also provide the first evidence of a neuronal localisation of melatonin in an invertebrate, which suggests that melatonin may act as a neurotransmitter or neurohormone in the least evolved animals endowed with a nervous system. PMID- 9336227 TI - Lectin histochemistry of the metathoracic ganglion of the locust Schistocerca gregaria before and after axotomy of the tympanal nerve. AB - The thoracic ganglia of insects exhibit a highly ordered organization. It seems possible that the information underlying the emergence of this order during development and its maintenance throughout insect life is given via a distinct pattern of molecules distributed within the ganglion. The question we asked was whether the adult insect ganglion is subdivided by the distribution of specific carbohydrates and furthermore whether or not this distribution changes during degeneration and regeneration of neurons. In order to determine the normal carbohydrate distribution, we stained sections of the intact metathoracic ganglion of the locust Schistocerca gregaria with fluorescence-coupled lectins. We succeeded in labeling three sensory neuropil areas with either peanut agglutinin (PNA): Phaseolus vulgaris erythrolectin (PVE), soybean agglutinin, wheat germ agglutinin (WGA), or Vicia villosa agglutinin. Apart from this, PNA, WGA, and succinylated WGA also selectively labeled some neuronal cell bodies, including dorsal unpaired median neurons. Datura stramonium lectin (DSL), Griffonia simplicifolia lectin II, and Solanum tuberosum lectin (STL) bound to glial cells or glia surrounding extracellular matrix. A few lectins stained all structures within the ganglion; some showed no binding at all. In the second part of our study, we tested whether carbohydrates were differentially regulated during transient deafferentation after the axotomy of the tympanal nerve. Binding of PNA and PVE within the auditory neuropil did not change. However, binding of the two glia-associated markers, DSL and STL, clearly differed from that found in intact animals; they bound transiently (day 3-4 until day 10-20 post-surgery) to axonal tracts and neuropils of the axotomized sensory afferents. PMID- 9336228 TI - Synaptic targets of cholinergic terminals in the pulvinar nucleus of the cat. AB - We compared the cholinergic innervation of the pulvinar nucleus, a thalamic association nucleus, to previous studies of the cholinergic innervation of the dorsal lateral geniculate nucleus (dLGN), a thalamic relay nucleus. Both nuclei receive a dense innervation from cholinergic cells of the brainstem parabrachial region (PBR). In the dLGN, PBR terminals are located in close proximity to retinal terminals. Our goal was to determine whether PBR terminals in the pulvinar nucleus are located in close proximity to corticothalamic terminals. We identified PBR terminals with a monoclonal antibody directed against choline acetyltransferase (ChAT). Cholinergic terminals contacted dendrites (142 of 160, or 89%) or vesicle-filled profiles (18 of 160, or 11%). A subset of 55 terminals was stained for gamma-aminobutyric acid (GABA) to determine whether profiles postsynaptic to cholinergic terminals originate from thalamocortical cells (GABA ) or interneurons (GABA+). The majority (44 of 55, or 80%) of postsynaptic profiles were GABA- dendrites. The minority (11 of 55, or 20%) were GABA+ dendrites with vesicles. This distribution of contacts is very similar to that seen in the dLGN. However, the most significant finding was that most cholinergic contacts (121 of 160, or 76%) were located within complex clusters identified as glomeruli. This is the primary site of contacts made by corticothalamic terminals originating from layer V cells. These results suggest that while the PBR enhances retinal signals in the dLGN, it may also enhance cortical signals in the pulvinar nucleus. Thus, activity in the PBR may stimulate both an increased flow of retinal information to visual cortex, as well as an increased flow of information between different visuomotor areas of cortex. PMID- 9336229 TI - Morphological correlates of neural regeneration in the feeding system of Aplysia californica after central nervous system lesions. AB - Morphological techniques were used to study regeneration of central neural pathways involved in feeding behavior following bilateral crushes of the cerebral buccal connectives (CBCs). Electron microscopic analysis revealed that CBC crushes completely transect axons within the nerve core while leaving a remnant of the nerve sheath intact. Changes in the ultrastructure of the CBCs at the crush site were determined for 1, 7, 14, 21, and 50 days postlesion. At 1 day postlesion, the crush site was no longer compressed, and the nerve core had assumed a circular shape. In addition, several small axon profiles were evident, and large areas of tissue debris and prominent microglial cells were observed. Membranous debris and hemocytes were also present in sinuses that appeared in the sheath adjacent to the crush site. From 7 to 50 days postlesion, the core of the nerve at the crush site increased in size due to the addition of small diameter axons. Initially, the sheath surrounding the crush site exhibited hyperplasia and contained a few small bundles of processes, apparently due to newly sprouted axons that had strayed from the nerve core. By 50 days postlesion, the crush site appeared nearly normal; the nerve core was reacquiring the normal radial pattern of axon profiles with some medium-sized axon profiles covered with glial sheath and exhibiting invaginations typical of the intact CBC. However, there was still a distinct lack of large diameter axons. Cobalt backfills across the crush site revealed neurons in the cerebral ganglion by postlesion day 9. Positions of stained cell bodies were consistent with those observed in controls, although the numbers of stained neurons did not recover to control levels even by postlesion day 63. The changes in the crush site and return of cell body staining with time postlesion are correlated with the recovery of consummatory feeding. PMID- 9336230 TI - Expression of cadherin-8 mRNA in the developing mouse central nervous system. AB - The expression of cadherin-8 was mapped by in situ hybridization in the embryonic and postnatal mouse central nervous system (CNS). From embryonic day 18 (E18) to postnatal day 6 (P6), cadherin-8 expression is restricted to a subset of developing brain nuclei and cortical areas in all major subdivisions of the CNS. The anlagen of some of the cadherin-8-positive structures also express this molecule at earlier developmental stages (E12.5-E16). The cadherin-8-positive neuroanatomical structures are parts of several functional systems in the brain. In the limbic system, cadherin-8-positive regions are found in the septal region, habenular nuclei, amygdala, interpeduncular nucleus, raphe nuclei, and hippocampus. Cerebral cortex shows expression in several limbic areas at P6. In the basal ganglia and related nuclei, cadherin-8 is expressed by parts of the striatum, globus pallidus, substantia nigra, entopeduncular nucleus, subthalamic nucleus, zona incerta, and pedunculopontine nuclei. A third group of cadherin-8 positive gray matter structures has functional connections with the cerebellum (superior colliculus, anterior pretectal nucleus, red nucleus, nucleus of posterior commissure, inferior olive, pontine, pontine reticular, and vestibular nuclei). The cerebellum itself shows parasagittal stripes of cadherin-8 expression in the Purkinje cell layer. In the hindbrain, cadherin-8 is expressed by several cranial nerve nuclei. Results from this study show that cadherin-8 expression in the embryonic and postnatal mouse brain is restricted to specific developing gray matter structures. These data support the idea that cadherins are a family of molecules whose expression provides a molecular code for the regionalization of the developing vertebrate brain. PMID- 9336231 TI - Functional circuitry in the brain of immune-challenged rats: partial involvement of prostaglandins. AB - This study investigated the role of prostaglandins (PGs) on the neuronal activity and the transcription of corticotropin-releasing factor (CRF) in the brain of conscious immune-challenged rats. Intravenous (i.v.) administration of indomethacin, an inhibitor of PG synthesis, was performed prior to and after the intraperitoneal (i.p.) injection of different doses [250 microg, 25 microg, and 2.5 microg/100 g body weight (b.w.)] of the immune activator lipopolysaccharide (LPS). Systemic administration of the high and middle doses of LPS caused a robust and widespread induction of both immediate-early genes (IEGs), c-fos and nerve growth factor-inducible gene B (NGFI-B) mRNAs, whereas injection of the low dose selectively triggered c-fos expression within the sensorial circumventricular organs. Pretreatment with indomethacin did not prevent c-fos transcription in the rat brains challenged with the high dose of LPS at 3 hours postinjection. Inhibition of PG formation was more effective for interruption of the neuronal activation in animals injected with 25 microg LPS/100 g b.w., although the influence depended on the structures and the groups of activated cells. Indeed, PG inhibition significantly altered LPS-induced c-fos mRNA expression in the medial preoptic area/organum vasculosum of the lamina terminalis, the periventricular nucleus, the paraventricular nucleus of the hypothalamus (PVN), and the ventrolateral medulla (VLM) but not in many other regions, including the subfornical organ, the central nucleus of the amygdala, the arcuate nucleus/median eminence, the parabrachial nucleus, the choroid plexus, and the nucleus of the solitary tract (NTS). In the hypothalamic PVN, inhibition of both c-fos and NGFI-B transcripts by indomethacin was also associated to an abolished influence of the endotoxin on the transcription of neuroendocrine CRF; induction of CRF primary transcript by the middle dose of LPS was selective to the PVN and was completely blocked by pretreatment with indomethacin. Moreover, a large number of tyrosine hydroxylase (TH) immunoreactive neurons of the VLM (A1/C1) and the NTS (A2/C2) were positive for c fos mRNA in immune-challenged rats, an effect that was largely prevented by indomethacin in the VLM but not in the NTS. These results indicate that the role of PGs in mediating the stimulatory influence of the acute-phase response depends on the severity of the systemic stressful situation, the brain regions, and the cell groups as well as the activated target genes. PMID- 9336232 TI - Identification of oligodendrocyte precursors in the myelinated streak of the adult rabbit retina in vivo. AB - Whole-mount techniques have been applied to the myelinated axons of the rabbit retina in order to study oligodendrocytes in the developing and adult central nervous system in vivo. Retinas from rabbits on embryonic day (E) 26 to postnatal day (P) 50 and from adults were subjected to immunohistochemistry with antibodies to glial fibrillary acidic protein (GFAP), vimentin, glycolipid O4, myelin basic protein (MBP), galactocerebroside (Gal-C), 4D6, and Griffonia simplicifolia isolectin, markers chosen for their specificity for astrocytes, cells of the oligodendrocyte lineage, Muller cells, and microglia. The first glial cells labelled within the myelinated streak (MS) were vimentin+ astrocytes. These cells were first apparent near the optic disc at E26. Their numbers and distribution increased markedly between E26 and E30, but between E29 and P3, vimentin expression was replaced by GFAP expression. Between P3 and P4, O4+, Gal-C-, MBP oligodendrocyte precursor cells, O4+, Gal-C-, MBP- pre-oligodendrocytes, and O4+, Gal-C-, MBP- immature oligodendrocytes appeared in low numbers in the region adjacent to the optic nerve head. O4-/+, Gal-C+, MBP+ mature oligodendrocytes appeared soon after at P4 to P5. With age, the outer extent of Gal-C and MBP immunoreactivity expanded, with positive cells forming a continuous sheath around nerve fibre bundles. The sequence of gliogenesis in the (MS) of the rabbit retina thus appears similar to that in the rat optic nerve. In adult rabbits, a large population of O4+ cells was distributed across the MS, the outer limit of the cells coinciding with the outer limit of retinal vessels. These O4+ cells could be classified into three stages of differentiation on the basis of expression of developmental markers and morphology: O4+, Gal-C-, MBP- oligodendrocyte precursor cells with a bipolar, unipolar, or simple morphology; O4+, Gal-C-, MBP- pre oligodendrocytes with a complex multipolar morphology; and O4+, Gal-C+, MBP- immature oligodendrocytes with a complex multipolar morphology. The final stage of differentiation was characterized as O4-, Gal-C+, MBP+ mature oligodendrocytes with multiple parallel processes aligned along nerve fibre bundles. These results provide in vivo evidence for the existence of a substantial population of oligodendrocyte precursor cells and non-myelin-producing immature oligodendrocytes in the MS of the adult rabbit retina. This preparation makes possible in situ patch clamping and determination of other functional responses of these cell types in vivo. PMID- 9336233 TI - Functional characterization of substance P receptors on cultured human spinal cord astrocytes: synergism of substance P with cytokines in inducing interleukin 6 and prostaglandin E2 production. AB - Following brain injury, astrocytes express receptors for cytokines and neuropeptides and secrete several regulatory mediators that have a well established role in inflammation, immunity, and tissue development or repair. To elucidate the role of substance P (SP), a neurotransmitter peptide of the tachykinin family, in inducing astrocyte secretory activities, we have examined the expression of SP receptors and the functional consequences of their activation in cultured astrocytes from the human embryonic brain or spinal cord. Radioligand binding studies revealed that only one type of SP receptors, the high affinity NK-1 receptor, was present on human astrocytes and that spinal cord astrocytes expressed about 6 times as many SP binding sites as brain astrocytes. Following SP treatment, a substantial inositol phosphate formation was observed in spinal cord astrocytes only. Stimulation of spinal cord astrocytes with SP alone did not induce secretion of cytokines [interleukin-6 (IL-6), granulocyte macrophage-CSF, macrophage chemoattractant protein-1 or leukemia inhibitory factor] or prostaglandin E2 (PGE2). Interestingly, however, SP selectively potentiated the inducing effect of IL-1beta on IL-6 and PGE2 secretion by spinal cord astrocytes without affecting the IL-1-beta-evoked secretion of other cytokines. SP also enhanced the small inducing effect of tumor necrosis factor alpha (TNF-alpha) on IL-6 and PGE2 secretion and that of transforming growth factor-beta on PGE2 secretion. These results suggest that SP can enhance immunoregulatory and neurotrophic astroglial functions mediated by IL-6 and PGE2 by acting in concert with a set of cytokines whose cerebral expression has been reported during development and in a variety of diseases. PMID- 9336234 TI - Metabotropic glutamate receptor agonists evoke calcium waves in isolated Muller cells. AB - Glutamate is the most prominent excitatory neurotransmitter in the retina and brain. It has become clear that the physiology of many glial cells, including retinal Muller cells, is modified by a host of neurotransmitters, including glutamate. The experiments presented here demonstrate that Muller cells isolated from the tiger salamander retina have metabotropic glutamate receptors that, when activated, lead to the release of calcium ions (Ca2+) from intracellular stores. The Ca2+-sensitive fluorescent dye, Fura-2, and video imaging microscopy were used to monitor changes in cytosolic calcium ion concentration ([Ca2+]i) evoked by glutamate (30-50 microM), (1S,3R)-ACPD (50-200 microM), quisqualate (10-50 microM), and L-AP4 (5-100 microM). Bath application of each of these metabotropic receptor agonists in the absence of extracellular Ca2+ resulted in an increase in [Ca2+]i that often began in the distal end of the cell and occurred later in the endfoot. This wavelike increase in [Ca2+]i is reminiscent of the Ca2+ waves evoked in these cells by other Ca2+ releasing agents such as ryanodine and caffeine. Extracellular application ofATP also evoked increases in [Ca2+] in Muller cells. The presence on Muller cells of receptors for retinal neurotransmitters, such as glutamate and ATP, demonstrates that these glial cells can respond to changes in the retinal extracellular environment and hence neuronal activity. Since Muller cells span almost all layers of the retina, they are likely to be exposed to most retinal neurotransmitters. The Ca2+ waves evoked in Muller cells by neurotransmitters could represent a form of signaling from the outer retinal layers to the inner ones. PMID- 9336235 TI - Deprenyl induces GFAP immunoreactivity in the intact and injured dopaminergic nigrostriatal system but fails to counteract axotomy-induced degenerative changes. AB - There is increasing evidence of a trophic-like mechanism for some effects ascribed to deprenyl therapy in the central nervous system. For that, we studied the effect of chronic treatment with deprenyl in an animal model of Parkinson's disease induced by unilateral knife transection of the medial forebrain bundle (MFB) in adult rats. The experimental conditions included a 3-week pretreatment with deprenyl before stereotaxic transection of the MFB. Following surgery, deprenyl treatment was maintained for 3 weeks. Neurochemical and immunohistochemical procedures were used to study the dopaminergic system and reactive astrocytes in the nigrostriatal system. Deprenyl treatment failed to counteract the axotomy-induced degenerative changes of the nigrostriatal dopaminergic system. However, it was effective in increasing the density of reactive astrocytes in terms of glial fibrillary acidic protein (GFAP) immunoreactivity in the intact contralateral substantia nigra and also in further enhancing the axotomy-induced increase of GFAP immunolabeled astrocytes in the lesioned substantia nigra. This deprenyl-induced effect on GFAP immunoreactivity was confined to substantia nigra without effect in striatum. In addition, we found a medial to lateral gradient decrease in the distribution pattern of GFAP immunolabeled astrocytes. Axotomy increased the number of reactive astrocytes in either striatal area examined, but yet the preferential distribution pattern of reactive astrocytes in striatum was still evident. PMID- 9336236 TI - Characterization of an inwardly rectifying chloride conductance expressed by cultured rat cortical astrocytes. AB - The biophysical and pharmacological properties of the inwardly rectifying Cl- conductance (IClh), expressed in rat type-1 neocortical cultured astrocytes upon a long-term treatment (1-3 weeks) with dibutyryl-cyclic-AMP (dBcAMP), were investigated with the whole-cell patch-clamp technique. Using intra- and extra cellular solutions with symmetrical high Cl- content and with the monovalent cations replaced with N-methyl-D-glucamine, time- and voltage-dependent Cl- currents were elicited in response to hyperpolarizing voltage steps from a holding potential of 0 mV. The inward currents activated slowly and did not display any time-dependent inactivation. The rising phase of the current traces was best fitted with two exponential components whose time constants decreased with larger hyperpolarization. The steady-state activation of IClh was well described by a single Boltzmann function with a half-maximal activation potential at - 62 mV and a slope of 19 mV that yields to an apparent gating charge of 1.3. The anion selectivity sequence was Cl- = Br- = I- > F- > cyclamate > or = gluconate. External application of the putative Cl- channel blockers 4,4 diisothiocyanatostilbene-2,2 disulphonic acid or 4-acetamido-4 isothiocyanatostilbene-2,2-disulphonic acid did not affect IClh. By contrast, anthracene-9-carboxylic acid, as well as Cd2+ and Zn2+, inhibited, albeit with different potencies, the Cl- current. Taken together, these results indicate that dBcAMP-treated cultured rat cortical astrocytes express a Cl- inward rectifier, which exhibits similar but not identical features compared with those of the cloned and heterologously expressed hyperpolarization-activated Cl- channel ClC 2. PMID- 9336238 TI - Astrocyte spreading in response to thrombin and lysophosphatidic acid is dependent on the Rho GTPase. AB - Astrocytes are typically star shaped cells playing diverse roles in the function of the nervous system. In astrocyte cultures established from the cerebral hemispheres of newborn rats, the cells have generally a polygonal fibroblast-like morphology, but acquire a stellate shape upon serum removal. When the serine protease thrombin or the bioactive lipid lysophosphatidic acid is added, the stellate cells revert to the flat morphology. Here we show that the effect of these agents is mediated via activation of the small GTP-binding protein Rho. Neither thrombin nor lysophosphatidic acid induced spreading of astrocytes microinjected with C3 transferase, an exoenzyme which ADP-ribosylates and thereby inactivates Rho. In contrast, the response of cells injected with a dominant negative form of Rac was unaffected. In addition, the injection of active Rho into stellate astrocytes mimicked the effect of thrombin and lysophosphatidic acid and an injection of C3 into flat cells grown in serum induced stellation. The conversion from a stellate to a spread morphology upon activation of Rho resulted in the formation of stress fibers and focal adhesions which most probably are key events in establishing and stabilizing the altered cytoarchitecture. These results suggest that Rho plays a crucial role in determining the shape of astrocytes and thereby may modulate their interaction with neurons in vivo. PMID- 9336237 TI - Localization of perlecan (or a perlecan-related macromolecule) to isolated microglia in vitro and to microglia/macrophages following infusion of beta amyloid protein into rodent hippocampus. AB - The origin of the heparan sulfate proteoglycan (PG), perlecan, in beta-amyloid protein (A beta)-containing amyloid deposits in Alzheimer's disease (AD) brain is not known. In the present investigation we used indirect immunofluorescence, SDS PAGE, and Western blotting with a specific perlecan core protein antibody to identify possible cell candidates of perlecan production in both primary cell cultures and in a rat infusion model. Double and triple-labeled indirect immunofluorescence was performed on dissociated primary rat septal cultures using antibodies for specific identification of cell types and for perlecan core protein. In mixed cultures of both embryonic day 18 (containing neurons and glia) and postnatal day 2-3 (devoid of neurons), microglia identified by labeling with OX-42 or anti-ED1 were the only cell type also double labeled with an affinity purified polyclonal antibody against perlecan core protein. Similar immunolabeling of microglia with the anti-perlecan antibody was also observed in purified cultures of post-natal rat microglia. Analyses of PGs from cultured postnatal rat microglia by Western blotting using a polyclonal antibody against perlecan core protein revealed an approximately 400 kDa band in cell layer, which was intensified following heparitinase/heparinase digestion, suggestive of perlecan core protein. Other lower Mr bands were also found implicating either degradation of the 400 kDa core protein or the presence of separate and distinct gene products immunologically related to perlecan. Reverse transcription followed by polymerase chain reaction using human perlecan domain I specific primers demonstrated perlecan mRNA in cultured human microglia derived from postmortem normal aged and AD brain. Following a 1-week continuous infusion of A beta (1-40) into rodent hippocampus, immunoperoxidase immunocytochemistry and double-labeled immunofluorescent studies revealed perlecan accumulation primarily localized to microglia/macrophages within the A beta infusion site. These studies have identified microglia/macrophages as one potential source of perlecan (or a perlecan-related macromolecule) which may be important for the ongoing accumulation of both perlecan and A beta in the amyloid deposits of AD. PMID- 9336239 TI - Increase of intracellular free Ca2+ in microglia activated by prion protein fragment. AB - A synthetic peptide consisting of amino acid residues 106 to 126 of the human prion protein (PrPc) that forms fibrils in vitro is toxic to cultured neurons. We have previously shown that the neurotoxic effect of this peptide is related to microglia activation (Brown et al., 1996a). For closer insight into this process of activation, we investigated the effect of the peptide on the intracellular free Ca2+ concentration ([Ca2+]i) in cultured microglia using Fura-2. Cultured microglia from wild-type as well as from PrPc gene-ablated mice (Prn-p0/0) responded to exposure to PrP106-126 with an increase in intracellular free calcium within 30 min. We observed two types of responses. Both in wild-type and Prn-p0/0 mice about half of the tested cells presented a small and often transient calcium increase after peptide application which was found to be independent of the extracellular calcium concentration. However, a further 33% of wild-type cells showed a strong and often permanent calcium increase depending on the extracellular calcium concentration, which was only rarely observed in Prn p0/0 cells. To determine whether the response depended on the activation state of the microglia, we also examined LPS-treated activated microglia. The character of the calcium response remained unchanged, but significantly fewer cells responded. Our findings demonstrate the earliest reaction of microglia to a PrP fragment known to date. PMID- 9336301 TI - Treatment of cirrhotic rats with L-ornithine-L-aspartate enhances urea synthesis and lowers serum ammonia levels. AB - CCl4-induced cirrhosis of rats was used for studying the influence of L-ornithine L-aspartate (OA) on hyperammonemia. OA given to cirrhotic rats (2 g/kg daily) for 2 wk slightly increased net body weight and led to a significant increase in plasma urea levels and a decrease in plasma ammonia levels. Serum concentrations of glutamate, glutamine and arginine decreased significantly. In the livers of the OA-treated rats the activities of carbamoylphosphate synthetase I and arginase increased by 30 and 40%, respectively, approaching normal levels. No change in the activities of the other urea cycle enzymes as well as of glutamate dehydrogenase, glutaminase and glutamine synthetase was found. The negative correlation between glutamine synthetase activity and plasma ammonia levels reported previously for cirrhotic rats (Gebhardt and Reichen, Hepatology 20:684 691, 1994) was corroborated for cirrhotic animals not treated with OA, but was no longer apparent in OA-treated cirrhotic rats. Despite this improvement, plasma ammonia levels still varied considerably reflecting the variable accessibility and activities of glutamine synthetase in cirrhotics. Cultured hepatocytes from the two groups of rats showed a similar stimulation of urea production by addition of ammoniumacetate and/or OA to Hanks' buffered salt solution. In Williams medium E, however, the hepatocytes from the OA group produced significantly more urea than those from controls. These results suggest that treatment of cirrhotic rats with OA considerably improves urea production favoring the detoxification of ammonia that, however, is still limited by the severe alterations in liver architecture that are not influenced by OA in a 2-wk period. PMID- 9336302 TI - Characterization of unconditioned behavioral effects of dopamine D3/D2 receptor agonists. AB - A series of experiments examined the ability of dopamine D3/D2 receptor agonists [(+)-(4aR,10bR)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]b enzopyrano-[4,3-b]-1,4 oxazin-9-ol hydrochloride (PD 128,907), (+/-)-7-hydroxy-dipropylaminotetralin hydrobromide (7-OH-DPAT), quinpirole and bromocriptine] to produce a variety of dopaminergically mediated behaviors. The effects of these drugs with selectivity for D3/D2 receptors over D1 receptors were compared with those produced by the selective D1 agonists [(+/-)-Phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8 diol hydrochloride (SKF 38393), (+/-)-6-Chloro-7,8-dihydroxy-3-allyl-1-phenyl 2,3,4,5-tetrahydro-1H-3-be nzazepine hydrobromide (SKF 82958)], a nonselective dopaminergic agonist (apomorphine), and an indirect dopamine agonist (cocaine). The D3/D2 agonists decreased locomotor activity, had no effect on gnawing and only inconsistently induced climbing in mice. Further, these agonists dose dependently produced scratching in squirrel monkeys. In contrast, the D1 agonists, SKF 82958 and SKF 38393, did not produce scratching in squirrel monkeys. Whereas the full D1 agonist, SKF 82958, produced increases in locomotor activity and in climbing and gnawing, the partial D1 agonist, SKF 38393, did not increase the frequencies of these behaviors. The nonselective dopamine agonist, apomorphine, produced decreases in locomotor activity and increases in climbing and gnawing in mice. Apomorphine dose-dependently produced scratching in squirrel monkeys. The indirect dopamine agonist, cocaine, produced increases in locomotor activity and climbing, but had no effect on climbing or gnawing in mice and did not produce scratching in squirrel monkeys. These findings suggest that D3/D2 agonists can be distinguished on various behavioral measures from the nonselective agonist, apomorphine (gnawing), D1 agonists (scratching) and the indirect agonist, cocaine (locomotor activity and scratching). Behaviors once attributed to stimulation of D2 (locomotor activity and scratching) or D1/D2 (climbing and gnawing) receptors may also involve dopamine D3 receptors. PMID- 9336303 TI - Comparative effects of two direct and indirect factor Xa inhibitors on free and clot-bound prothrombinase. AB - Factor Xa, as with thrombin, binds to the clot and contributes to the propensity of thrombi to activate the coagulation system. The aim of this work was to compare the extent of prothrombinase inhibition produced by two factor Xa inhibitors: the antithrombin III-dependent synthetic pentasaccharide (SR 90107/Org 31540) and DX-9065A, a direct factor Xa inhibitor. When incubated together with prothrombin, factor Xa, phospholipids, antithrombin III and calcium, clots formed from human plasma exhibited a prothrombinase activity as measured through fragment 1-2 (F1+2) generation. Ten washes of the clot were required to achieve complete removal of unbound factor Xa. The absence of F1+2 generation brought about by washed clots in buffer when factor V was omitted, or in the presence of annexin V, indicated that they contained bound factor Xa and phospholipids but no factor V/Va. In all tested experimental conditions, clot bound-factor Xa-induced F1+2 generation was inhibited by SR 90107/AT and DX-9065A with IC50 in the same range of concentrations (0.5 microM). In contrast, the inhibition of prothrombinase formed with factor Xa, factor Va phospholipids and calcium in buffer was observed at significantly lower concentrations of DX-9065A than of SR 90107/AT (respective IC50 concentrations: 0.1 and 70 microM). In vivo, fibrin accretion onto a preformed thrombus as well as venous thrombosis induced in the jugular vein of rabbits was inhibited by SR 90107 and DX-9065A in the same range of concentrations therefore showing that inhibition of clot-bound factor Xa is a predominant factor for the antithrombotic activity of both direct and indirect inhibitors for factor Xa. PMID- 9336304 TI - Motor responses in rat ileum evoked by nitric oxide donors vs. field stimulation: modulation by pituitary adenylate cyclase-activating peptide, forskolin and guanylate cyclase inhibitors. AB - This study examines nitric oxide (NO) mediated effects on longitudinal muscle with adherent myenteric ganglia from rat ileum in vitro using NO donors and electrical field stimulation. Electrical field stimulation (20 Hz) caused a biphasic response-a relaxation followed by a contraction. NG-nitro-L-arginine methyl ester almost totally abolished the relaxation and L-arginine restored it. The contraction was unaffected. The NO donors sodium-nitroso-N acetylpenicillamine (SNAP) and sodium-nitroprusside also induced a biphasic response, a contraction followed by relaxation. Relaxations mediated by neuronally released NO were not blocked by methylene blue or 1H [1,2,4]oxadiazolo[4,3-a]-quinoxalin-1-one suggesting that they are independent of a rise in intracellular cyclic guanylate cyclase. Their amplitude was unaffected by forskolin. The relaxations evoked by NO (or a NO-related substance) liberated from SNAP were blocked by methylene blue or 1H-[1,2,4]oxadiazolo[4,3-a] quinoxalin-1-one indicating a cyclic guanylate cyclase-dependent mechanism of action. Pituitary adenylate cyclase-activating peptide and forskolin, but not vasoactive intestinal peptide or neuropeptide Y, caused a marked leftward shift of the concentration-response curve of the SNAP-induced relaxation. The contractions induced by SNAP were blocked by methylene blue and 1H [1,2,4]oxadiazolo[4,3-a]-quinoxalin-1-one and thus, cyclic guanylate cyclase dependent. The SNAP-induced contractions were abolished by pituitary adenylate cyclase-activating peptide and forskolin, but unaffected by vasoactive intestinal peptide or NPY. In conclusion, motor responses evoked by NO released from NO donors vs. neuronally released NO reveals different mechanisms of action. PMID- 9336305 TI - The relationship between reinforcing effects and in vitro effects of D1 agonists in monkeys. AB - The reinforcing effects of many psychomotor stimulants have been related to increased dopaminergic neurotransmission and stimulation of central nervous system (CNS) dopamine (DA) receptors. Consistent with this notion, some drugs that directly stimulate DA receptors have been found to function as positive reinforcers. The present experiments were designed to examine why some, but not all, D1 receptor agonists can function as reinforcers in rhesus monkeys by comparing behavioral and CNS in vitro measures of potency and efficacy. Seven rhesus monkeys were allowed to self-administer cocaine under a progressive-ratio (PR) schedule until stable responding was established. Various doses of D1 agonists, previously reported to function as positive reinforcers, were then made available for self-administration. Stimulation of cAMP production in rhesus and rat striatal tissue was studied for these compounds and for D1 agonists previously reported not to function as positive reinforcers in monkeys (SKF 38393, SKF 77434 and S(-)-6-BrAPB). Blockade of DA uptake in rat striata was also examined for all compounds. SKF 81297, SKF 82958 and R(+)-6-BrAPB maintained responding under the PR schedule and did not differ significantly in efficacy as positive reinforcers; SKF 81297 was less potent than the other two agonists. SKF 81297, SKF 82958 and R(+)-6-BrAPB stimulated higher levels of cAMP production in rhesus striata than did SKF 38393, SKF 77434 and S(-)-6-BrAPB. In contrast, all compounds blocked DA uptake. Thus, reinforcing efficacy among D1 agonists increases with efficacy in stimulating D1 receptors. PMID- 9336306 TI - The digoxin-propafenone interaction: characterization of a mechanism using renal tubular cell monolayers. AB - When propafenone is given with digoxin, digoxin serum concentrations increase. Although the digoxin-propafenone interaction is well known clinically, the mechanism by which propafenone interferes with digoxin elimination is unclear. To test the hypothesis that propafenone or one or both of its two major metabolites, 5-hydroxypropafenone (5-OHP) and N-depropylpropafenone (NDPP), inhibit the P glycoprotein-mediated net renal tubular secretion of digoxin, we examined the transport of digoxin and the well-studied P-glycoprotein substrate vinblastine across confluent Madin-Darby canine kidney cell monolayers in the absence and presence of propafenone, 5-OHP and NDPP. Propafenone and its two major metabolites significantly inhibit the secretory flux of digoxin and vinblastine (propafenone > 5-OHP >> NDPP). Despite decreases in net transport, cellular digoxin accumulation did not decrease, suggesting that neither propafenone nor its metabolites prohibited digoxin from entering the cells at the basolateral side. NDPP, but not 5-OHP, was detected after 48 hr of incubation of the cells with propafenone alone. When the cells were incubated with propafenone or 5-OHP, apical accumulation of 5-OHP, but neither propafenone nor NDPP, against a concentration gradient was observed. These findings are consistent with the hypothesis that the digoxin-propafenone interaction results from the inhibition of the renal tubular transport of digoxin by propafenone and its metabolites. Our data suggest that propafenone is an inhibitor of P-glycoprotein, whereas 5-OHP is a possible substrate. PMID- 9336308 TI - Ropivacaine inhibits leukocyte rolling, adhesion and CD11b/CD18 expression. AB - Ropivacaine, a new local anesthetic, is currently being investigated for the treatment of ulcerative colitis, with promising results so far. The aim of this study was to examine anti-inflammatory properties of ropivacaine with regard to its effects on vascular permeability and inflammatory leukocyte behavior in vivo. The effects on leukocyte rolling, firm adhesion and vascular permeability were examined in the hamster cheek pouch microvasculature via intravital microscopy, and the effects on leukocyte adhesion molecules were examined in vitro by means of flow cytometry. In large venules, leukocyte adhesion induced by topical leukotriene B4 (LTB4) was almost completely inhibited during the combined application of ropivacaine and LTB4. The spontaneous rolling leukocyte flux was reduced by 72%, the rolling leukocyte fraction by 47% and the total leukocyte flux, which reflects blood flow, by 47%. In postcapillary venules, ropivacaine abolished rolling and LTB4-induced firm adhesion of leukocytes. LTB4 challenge also resulted in increased plasma exudation that was almost completely inhibited by ropivacaine. Moreover, ropivacaine inhibited the tumor necrosis factor alpha induced up-regulation of CD11b/CD18 and L-selectin shedding by human leukocytes in vitro. Our results suggest that ropivacaine exerts anti-inflammatory activity, and this appears to be mediated to a significant extent by inhibition of both leukocyte rolling and adhesion. PMID- 9336307 TI - The prediction of human pharmacokinetic parameters from preclinical and in vitro metabolism data. AB - We describe a comprehensive retrospective analysis in which the abilities of several methods by which human pharmacokinetic parameters are predicted from preclinical pharmacokinetic data and/or in vitro metabolism data were assessed. The prediction methods examined included both methods from the scientific literature as well as some described in this report for the first time. Four methods were examined for their ability to predict human volume of distribution. Three were highly predictive, yielding, on average, predictions that were within 60% to 90% of actual values. Twelve methods were assessed for their utility in predicting clearance. The most successful allometric scaling method yielded clearance predictions that were, on average, within 80% of actual values. The best methods in which in vitro metabolism data from human liver microsomes were scaled to in vivo clearance values yielded predicted clearance values that were, on average, within 70% to 80% of actual values. Human t1/2 was predicted by combining predictions of human volume of distribution and clearance. The best t1/2 prediction methods successfully assigned compounds to appropriate dosing regimen categories (e.g., once daily, twice daily and so forth) 70% to 80% of the time. In addition, correlations between human t1/2 and t1/2 values from preclinical species were also generally successful (72-87%) when used to predict human dosing regimens. In summary, this retrospective analysis has identified several approaches by which human pharmacokinetic data can be predicted from preclinical data. Such approaches should find utility in the drug discovery and development processes in the identification and selection of compounds that will possess appropriate pharmacokinetic characteristics in humans for progression to clinical trials. PMID- 9336309 TI - Calcium-dependent mitochondrial formation of species mediating DNA single strand breakage in U937 cells exposed to sublethal concentrations of tert butylhydroperoxide. AB - Treatment of U937 cells with a sublethal albeit DNA-damaging concentration of tert-butylhydroperoxide (tB-OOH) enhanced mitochondrial Ca++ uptake and ruthenium red (RR), a polycation that inhibits the calcium uniporter of mitochondria, significantly reduced the extent of DNA cleavage generated by the hydroperoxide. Release of Ca++ from the ryanodine(Ry)/caffeine(Cf)-sensitive stores further increased mitochondrial Ca++ uptake and elicited a parallel enhancement in DNA strand scission induced by tB-OOH that was prevented by both Ry and RR. DNA damage caused by tB-OOH alone or associated with either Cf or RR was prevented by iron chelators, insensitive to antioxidants and repaired with kinetics superimposable with those observed after treatment with H2O2. Cf enhanced the DNA damaging effects of tB-OOH in permeabilized cells as well, and similar effects were observed upon addition of CaCl2. Cf did not further increase the formation of DNA lesions elicited by tB-OOH in the presence of CaCl2. The enhancing effects of Cf were prevented by RR and ryanodine, whereas those mediated by exogenous calcium were prevented only by RR. DNA strand scission caused by tB-OOH alone or associated with Cf in the permeabilized cell system was severely inhibited by ethylene glycol-bis(beta-aminoethyl ether)-N, N,N',N'-tetraacetic acid. The mechanism(s) whereby Ca++ promotes the mitochondrial formation of species that will ultimately result in the formation of DNA lesions was subsequently analyzed using intact as well as permeabilized cells. Hydrogen peroxide was identified to be one of these species. PMID- 9336310 TI - Contribution of constrictor prostanoids to the calcium-dependent basal tone in the aorta from rats with aortic coarctation-induced hypertension: relationship to nitric oxide. AB - Rings of thoracic aortae taken from rats made hypertensive by aortic coarctation express a calcium-dependent basal tone. We investigated whether this basal tone is mediated by prostanoids. To this end, we contrasted the effects of indomethacin, an inhibitor of cyclooxygenase, and of ifetroban, an antagonist of thromboxane A2/prostaglandin endoperoxide H2 receptors, on basal tone in aortic rings taken from normotensive and hypertensive rats. Rings with endothelium from normotensive rats were unaffected by indomethacin and ifetroban. However, in endothelium-intact rings from hypertensive rats, the basal tone was reduced 65 to 75% by indomethacin and ifetroban, but not by CGS13080, an inhibitor of thromboxane synthase. The reductions in tone elicited by indomethacin and ifetroban in rings from hypertensive rats were eliminated upon removal of the endothelium and were attenuated when the rings were pretreated with an inhibitor of nitric oxide synthase (N omega-nitro-L-arginine methyl ester or N omega-nitro L-arginine) or an inhibitor of soluble guanylate cyclase. Neither indomethacin nor ifetroban affected tissue cGMP levels or nitrite release in aortic rings taken from hypertensive rats. However, sodium nitroprusside offset the inhibitory effects of N omega-nitro-L-arginine methyl ester, on the relaxant responses to indomethacin and ifetroban. These data suggest that a constrictor prostanoid other than thromboxane A2, presumably prostaglandin endoperoxide H2 contributes to the implementation of the basal tone in rings from hypertensive rats and that part of the relaxant response to indomethacin and ifetroban is linked to nitric oxide. PMID- 9336311 TI - The "kynurenate test", a biochemical assay for putative cognition enhancers. AB - Some putative cognition enhancers (oxiracetam, aniracetam and D-cycloserine) were previously shown to prevent the kynurenic acid antagonism of the N-methyl-D aspartate (NMDA)-evoked norepinephrine (NE) release in rat hippocampal slices. This functional in vitro assay was further characterized in the present work. D Serine, a glutamate coagonist at the NMDA receptor glycine site, concentration dependently (EC50 approximately 0.1 microM) prevented the kynurenate (100 microM) block of the NMDA (100 microM)-evoked [3H]NE release. L-Serine was ineffective up to 10 microM. The gamma-aminobutyric acidB (GABA[B]) receptor antagonist CGP 36742, reported to improve cognitive performance, potently prevented the kynurenate antagonism. The activity of CGP 36742 (1 microM) appeared to be unaffected by 10 microM (-)-baclofen, a GABA(B) receptor agonist; furthermore, CGP 52432, a GABA(B) antagonist more potent than CGP 36742, but reportedly devoid of nootropic properties, was inactive in the "kynurenate test." The novel putative cognition enhancer CR2249, but not its enantiomer CR2361, also potently prevented the kynurenate antagonism. In contrast, linopirdine, nicotine and tacrine were inactive. In rat hippocampal synaptosomes glycine and D-cycloserine enhanced the NMDA-evoked [3H]NE release, whereas oxiracetam and CR2249 did not. These four compounds were all similarly effective in preventing kynurenate antagonism, both in slices and in synaptosomes. The NMDA potentiation caused by glycine (0.1-100 microM) was not affected by 100 microM oxiracetam, which suggested that drugs active in the "kynurenate test" may bind to sites different from the glycine site of the NMDA receptor. To conclude, the "kynurenate test" is an in vitro assay useful in the identification and characterization of putative cognition enhancers acting via NMDA receptors. PMID- 9336313 TI - Effects of noradrenergic lesions on MPTP/MPP+ kinetics and MPTP-induced nigrostriatal dopamine depletions. AB - Norepinephrine (NE) depletion caused by damage to locus ceruleus neurons was shown to worsen experimental Parkinsonism induced by the neurotoxin 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) in monkeys and in rodents. However, it is not clear whether the lesion to the NE system enhances neurotoxicity in the nigrostriatal dopaminergic (DA) pathway and/or impairs the recovery of DA neurons once the neurotoxic insult is generated. In this study, we provide evidence that the lesion of NE terminals, induced by the selective neurotoxin N-(-2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4; 50 mg/kg), must occur before MPTP (30 mg/kg) administration in order to enhance MPTP toxicity. As a second step, we evaluated the acute effects of MPTP on the nigrostriatal DA pathway in NE lesioned animals compared with intact animals. We observed a more marked acute DA depletion, persisting at 12 h, in DSP-4 + MPTP-treated mice compared with MPTP injected controls. These findings, combined with the lack of an MPTP enhancement when NE depletion was induced 12 h after MPTP administration, suggest that in NE depleted animals, a more pronounced acute neuronal sensitivity to MPTP occurs. In line with the hypothesis of an acute protective effect by NE axons, we evaluated whether the enhancement of MPTP toxicity in NE-lesioned animals is achieved through alterations to the kinetics of MPTP and its metabolite. Our findings indicate that despite the pivotal role of NE terminals in taking up and storing 1 methyl-4-phenylpyridinium (MPP+), MPTP enhancement does not depend on modifications in the striatal kinetics of MPTP/MPP+ measured at seven different time intervals after MPTP administration. PMID- 9336312 TI - Antithrombotic efficacy of a recombinant nematode anticoagulant peptide (rNAP5) in canine models of thrombosis after single subcutaneous administration. AB - We describe the antithrombotic effects of recombinant nematode anticoagulant peptide (rNAP5), a selective and direct factor Xa inhibitor, after a single s.c. administration in canine models of arterial and venous thrombosis. The systemic anticoagulant effects of rNAP5 were evaluated initially in conscious dogs after s.c. dosing (0.03, 0.1 and 0.3 mg/kg) that resulted in a dose-dependent increase in the activated clotting time and the activated partial thromboplastin time. The antithrombotic effects of rNAP5 were evaluated in anesthetized dogs where saline or rNAP5 (0.03, 0.1 and 0.3 mg/kg s.c.) was administered 1 hr before the left circumflex coronary artery was subjected to electrolytic injury. In the saline group (n = 10), the left circumflex artery occluded in 79 +/- 9 min, and 5 of 10 animals progressed to sudden death due to ventricular fibrillation. rNAP5 significantly prolonged the time to occlusion in the 0.03 mg/kg (163 +/- 62 min) and 0.1 mg/kg (327 +/- 62) treatment groups (n = 6). In the 0.3 mg/kg group (n = 5), all of the injured vessels remained patent for 8 hr. There was a dose dependent reduction in the thrombus mass in the rNAP5-treated animals as compared with controls, as well as a lower mortality rate. rNAP5, in the doses of 0.03 and 0.1 mg/kg, did not alter the bleeding time, whereas 0.3 mg/kg produced a 5-fold increase. In a separate study, we evaluated the efficacy of rNAP5 (0.1 mg/kg) in the prevention of carotid artery and jugular vein thrombosis. In response to endothelial injury, the carotid artery and jugular vein in the saline group (n = 6) occluded in 142 +/- 16 and 100 +/- 11 min, respectively, compared with rNAP5, which maintained vessel patency in the carotid artery (6/6) and jugular vein (5/6) and significantly decreased the thrombus weights. The results demonstrate that rNAP5 has antithrombotic efficacy in canine models of arterial and venous thrombosis after a single s.c. administration. PMID- 9336314 TI - Nonlinear intestinal absorption of 5-hydroxytryptamine receptor antagonist caused by absorptive and secretory transporters. AB - The mechanism of the nonlinear concentration dependence of intestinal absorption of the 5-hydroxytryptamine receptor antagonist azasetron was studied by use of rat in situ intestinal perfusion, as well as an in vitro Ussing-type chamber method mounted with rat intestinal tissue and cultured monolayers of human adenocarcinoma Caco-2 cells. The intestinal absorption rate constant of azasetron evaluated by the Doluisio method increased significantly with increasing concentration of azasetron up to 10 mM in a nonlinear fashion and tended to decrease at higher concentrations. Mucosal-to-serosal directed permeation of [14C]azasetron across rat ileal sheets evaluated by the in vitro Ussing-type chamber method also increased in a nonlinear fashion in a low concentration range, followed by a decrease as the concentration was further increased, whereas serosal-to-mucosal directed permeation decreased in a concentration-dependent manner. Vectorial transport of [14C]azasetron across a Caco-2 cell monolayer was observed, with higher transport in the basolateral-to-apical direction at a trace concentration of azasetron. When the initial uptake rate of azasetron by Caco-2 cells was measured, it was saturable with an apparent half-saturation concentration of 15 mM and was reduced in the presence of several cationic compounds. These observations suggest that azasetron is taken up by a carrier mediated transport mechanism across the intestinal epithelial cells. When the steady-state uptake of [14C]azasetron was measured, it was increased in the presence of unlabeled azasetron and ondansetron. In addition, the steady-state uptake was enhanced in the presence of a P-glycoprotein inhibitor, cyclosporin A, and by ATP-depletion of the cells, although these treatments had no effect on the initial uptake of [14C]azasetron. Furthermore, the multidrug-resistant cancer cell line K562/ADM that overexpresses P-glycoprotein accumulated azasetron less extensively than did the parental drug-sensitive K562 cells. These results strongly suggest that azasetron is secreted into the intestinal lumen predominantly by P-glycoprotein. We conclude that intestinal transport of azasetron involves specialized transporters in both the absorptive and secretory directions, and the complex nonlinear intestinal absorption characteristics can be ascribed to the participation of multiple transport mechanisms. PMID- 9336315 TI - Estrogen and selective estrogen receptor modulator LY117018 enhance release of nitric oxide in rat aorta. AB - We report on the modulatory effects of chronic subcutaneous or oral estrogen and LY117018, a selective estrogen receptor modulator, on the release of nitric oxide in rings of rat aorta studied under isometric conditions. Dilator responses to acetylcholine (ACh; 10[8] to 10[-5] M) were obtained in phenylephrine (PE; 2 microM)-contracted aorta, and constrictor dose-response curves to PE (10[-8] to 10[-5] M) were generated before and after pretreatment with N omega-nitro-L arginine methyl ester (L-NAME; 200 microM), an inhibitor of nitric oxide synthase. Tissue segments were obtained from five groups of rats implanted with a subcutaneous pellet delivery system for 21 days: (1) male, (2) sham-operated placebo-treated female, (3) ovariectomized placebo-treated, (4) ovariectomized, 17beta-estradiol treated (0.5 mg/pellet) and (5) ovariectomized, progesterone (15 mg/pellet) and 17beta-estradiol (0.5 mg/pellet)-treated. Aortic rings from sham rats and ovariectomized rats receiving estrogen relaxed more to ACh (10[-6] to 10[-5] M) than did the rings from ovariectomized, progesterone plus estrogen treated and male rats (P < .05). They were also characterized by a greater potentiation of the PE responses after L-NAME compared with male, progesterone plus estrogen-treated and ovariectomized rats (P < .05) and a similar sensitivity to PE. In addition, ACh-induced relaxation and L-NAME-induced potentiation of PE contractions in aortic rings from rats dosed orally with LY117018 were similar to responses of aortic rings from rats dosed orally with estrogen. These results demonstrate that chronically administered estrogen and LY117018 enhance the release of nitric oxide from endothelium in rat aortic rings. PMID- 9336317 TI - Effects of tyrosine kinase inhibitors on antigen challenge of guinea pig lung in vitro. AB - The present study was conducted to examine the effects of two protein tyrosine kinase inhibitors, genistein and tyrphostin 47, on an in vitro model of allergic asthma. Guinea pigs were sensitized with purified IgG raised against ovalbumin (OA). Isolated sensitized bronchial rings contracted in response to OA in a concentration-dependent manner, maximum contraction being achieved at 1 microg/ml. Genistein and tyrphostin 47 concentration-dependently (10-100 microM) inhibited OA-induced anaphylactic contraction of the bronchi, as well as release of histamine and peptidoleukotrienes from chopped lung preparations. Genistein, but not tyrphostin 47, significantly suppressed bronchial contraction to leukotriene D4 at 50 microM and to histamine at 100 microM. Daidzein, an inactive congener of genistein, did not alter OA-induced anaphylactic contraction. However, it slightly reduced bronchial contraction to leukotriene D4 and the OA stimulated release of peptidoleukotrienes. The inhibitory effects were significantly weaker than those of genistein. Taken together, our results show that tyrphostin 47 inhibited anaphylactic contraction mainly by preventing mast cell degranulation, whereas genistein exerted inhibitory effects partly by blocking mast cell degranulation and partly by attenuating leukotriene D4-induced bronchial contraction. These findings suggest that protein tyrosine kinase inhibitors have a therapeutic potential as mast cell stabilizers in the treatment of allergic diseases such as bronchial asthma. PMID- 9336318 TI - Use of a nitronyl nitroxide to discriminate the contribution of nitric oxide radical in endothelium-dependent relaxation of control and diabetic blood vessels. AB - Nitronyl nitroxides react with nitric oxide radical (.NO) to form imino nitroxides. We used a nitronyl nitroxide, [2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl 3 oxide] (CPTIO) to evaluate the contribution of .NO to basal tone and acetylcholine-induced endothelium-dependent relaxation in control vs. diabetic rat aortic rings. In rings precontracted with phenylephrine, CPTIO produced an additional increment in tension that was greater in control vs. diabetic rings. Tension after CPTIO was similar to that observed in rings pretreated with the NO synthase inhibitor, L-nitroarginine or in rings without endothelium. This increment was insensitive to indomethacin, cysteine, tetraethylammonium or catalase, but was sensitive to inhibition by the soluble guanylate cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxaline-1-one. L Nitroarginine blocked relaxation to ACH by 100 and 90% in control and diabetic rings, respectively. In contrast, CPTIO produced a concentration-dependent inhibition of ACH-induced relaxation that was greater in control rings. The residual CPTIO-resistant component of relaxation was equivalent to 26 and 43% of initial precontraction in control vs. diabetic rings, respectively, and was not altered by indomethacin, catalase, cysteine or tetraethylammonium but was significantly inhibited by 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxaline-1-one. These data suggest the release of additional unknown factor(s) that cannot be discerned using NO synthase inhibitors only. This CPTIO-resistant dilator is likely not a cyclooxygenase product or a hyperpolarizing factor but a factor that acts, in part, by activation of guanylate cyclase. This substance is possibly .NO that is not available for reaction with CPTIO either by its diffusibility and sequestration or molecular rearrangement to a redox active form (i.e., not free .NO) or is a completely different vasodilator. The use of a more lipid soluble nitronyl nitroxide derivative suggests a portion of the CPTIO-resistant relaxation in diabetic (but not control) rings could be explained by .NO sequestered in the lipid phase. PMID- 9336316 TI - Molecular determinants of monosodium urate crystal-induced murine peritonitis: a role for endogenous mast cells and a distinct requirement for endothelial-derived selectins. AB - Injection of monosodium urate (MSU) crystals, the etiological cause of gouty arthritis, into murine peritoneal cavities produced an intense recruitment of polymorphonuclear leukocytes (PMN). After 3 mg MSU crystal injection, cell influx was maximal (approximately 10 x 10[6] cells per mouse) at 6 hr postinjection and sustained up to the 24 hr time-point. In mice depleted of mast cells by administration of compound 48/80 72 hr before challenge with MSU crystals a lower PMN influx was measured (58% reduction). The occurrence of endogenous mast cell activation, in the MSU response, was validated by the observation that MSU challenge reduced by more than 90% the number of intact mast cells recovered in the peritoneal washes. Pretreatment of mice with a histamine H1 antagonist (tripolidine; 0.5 mg/kg) or a platelet-activating factor receptor antagonist (WEB2086; 10 mg/kg) significantly reduced by 50 to 60% the number of PMN recovered from the peritoneal cavities. The molecular determinants of this process of leukocyte recruitment were also investigated. Treatment of mice with an anti-CD62P or anti-CD62E monoclonal antibody (mAb; 100 microg i.v.) produced a distinct inhibition of PMN recruitment measured at 6 hr, whereas only a combined administration of both monoclonal antibodies was effective in reducing by 60% the influx of PMN caused by the MSU crystals within 24 hr. In conclusion, these data highlight a role for endogenous mast cells and for endothelial-derived selectins in MSU crystal-induced PMN recruitment into the peritoneal cavity, and may be useful to dissect molecular mechanism(s) which may be operating in gouty arthritis. PMID- 9336319 TI - Block of IKs by the diuretic agent indapamide modulates cardiac electrophysiological effects of the class III antiarrhythmic drug dl-sotalol. AB - Indapamide is a diuretic agent with direct electrophysiological effects on ionic currents involved in cardiac repolarization. In particular, indapamide blocks the slow component of delayed rectifier potassium current. In contrast, most class III antiarrhythmic agents, such as dl-sotalol, block the rapid component of delayed rectifier potassium current. Computer simulations have suggested potentiation of drug effects on cardiac repolarization by the combined block of the rapid component of delayed rectifier potassium current and the slow component of delayed rectifier potassium current. Therefore, the objective of our study was to evaluate the modulation of cardiac electrophysiological effects of dl-sotalol by indapamide. Two indices of cardiac repolarization, monophasic action potential duration at 90% repolarization and effective refractory period, at two basic cycle lengths (800 and 400 msec) were determined in 24 anesthetized open-chest dogs. In two treatment groups (n = 6/group), data were obtained at base line and every 2 min during steadily increasing concentrations of dl-sotalol (0-40 microg/ml) either alone or in the presence of indapamide (500 ng/ml). Data were also obtained in dogs receiving either a low-dose (500 ng/ml) or a high-dose (up to 7.5 microg/ml) infusion regimen of indapamide alone. Administration of dl sotalol was associated with concentration-dependent increases in monophasic action potential duration at 90% repolarization and effective refractory period, whereas repolarization was only slightly altered by the administration of indapamide alone. However, concentration-response curves of dl-sotalol were shifted to the left in dogs treated with the combination of dl-sotalol and indapamide, and the EC50 values of dl-sotalol estimated for the prolongation of monophasic action potential duration at 90% repolarization and effective refractory period were decreased 3-fold during the coadministration of both drugs (P < .05 vs. dl-sotalol alone). Thus, under conditions of normal K+ levels, clinically relevant concentrations of indapamide modulate dl-sotalol effects on cardiac repolarization. Additional block of cardiac K+ currents, especially the rapid component of delayed rectifier potassium current and the slow component of delayed rectifier potassium current could explain these observations. PMID- 9336320 TI - In vivo metabolism-based discovery of a potent cholesterol absorption inhibitor, SCH58235, in the rat and rhesus monkey through the identification of the active metabolites of SCH48461. AB - SCH48461 is a selective and highly potent inhibitor of cholesterol absorption. In rats, SCH48461 is rapidly and completely metabolized in the first pass through the body. To compare the activity of the metabolites of SCH48461 with SCH48461 itself, an intestinally cannulated, bile duct-cannulated rat model for cholesterol absorption was developed. SCH48461 inhibited the absorption of cholesterol by 70%, whereas bile containing the metabolites of SCH48461 (henceforth, "metabolite bile") inhibited the absorption by greater than 95%. Very little of the recovered radioactive dose of SCH48461 was located in the intestinal lumen (7%) or wall (4%), whereas 85% appeared in bile. However, in rats treated with metabolite bile, 62% of the dose remained in the lumen, 13% was associated with the wall and only 24% reappeared in bile, which suggests that the activity of the metabolite bile may be related to its higher retention in the intestinal wall. Rats treated with metabolite bile had 64% and 84% less drug related radioactivity in their plasma and livers, respectively, compared with animals treated with SCH48461, which indicates that the metabolites are systemically less available than SCH48461. The metabolites in bile were separated by high-performance liquid chromatography; the most active fraction in the bile duct-cannulated rat model was identified by mass spectrometry as the glucuronide of the C4-phenol of SCH48461. The other fractions had moderate or no activity. Through the identification of the most active biliary metabolites of SCH48461 in the rat, we have discovered SCH58235, a novel cholesterol absorption inhibitor which is 400 times more potent than SCH48461 in the cholesterol-fed rhesus monkey. PMID- 9336321 TI - Cocaine and alcohol interactions in humans: neuroendocrine effects and cocaethylene metabolism. AB - The effects of 100 mg of intranasal cocaine in acute alcohol intoxication (0.8 g/kg) were evaluated in eight experienced and nondependent healthy volunteers in a double-blind double-dummy, controlled, randomized, crossover clinical study. The combination of alcohol and cocaine produced greater increases in HR, rate pressure product and pleasurable-related subjective effects (euphoria, well being) compared with the effects of cocaine. The drug combination reduced the alcohol-induced sedation, but feelings of drunkenness were not significantly counteracted. Cardiovascular changes induced by the combination condition caused an increase in myocardial oxygen consumption that may be related to an increased risk of cardiovascular toxicity. The augmented subjective euphoria may explain why the drug combination is more likely to be abused than is cocaine or alcohol alone. Plasma cortisol concentrations were significantly higher after concomitant alcohol and cocaine use than with cocaine alone. The administration of cocaine did not alter alcohol-induced hyperprolactinemia. Although cocaine produced a slight decrease in plasma concentrations of prolactin when administered alone, it did not antagonize the effects of alcohol on prolactin secretion when alcohol and cocaine were given simultaneously. The combination increased cocaine and norcocaine plasma concentrations, and induced the synthesis of cocaethylene and norcocaethylene. The enhancement of cocaine effects in the drug combination may be due to initially increased cocaine plasma levels followed by the additive effect of cocaethylene, although a pharmacodynamic interaction could not be excluded. PMID- 9336322 TI - Metabolism of cAMP to adenosine in the renal vasculature. AB - We recently demonstrated that cAMP added to the perfusate increased the renal venous recovery of adenosine in the isolated rat kidney, an effect blocked by inhibition of ecto-phosphodiesterase and ecto-5'-nucleotidase. Although our previous study established the cAMP-adenosine pathway, i.e., the conversion of cAMP to adenosine, as a viable metabolic pathway within the kidney, that study did not determine whether conversion of arterial cAMP to adenosine recoverable in the venous effluent occurred in the tubules versus nontubular sites. In the current study, we addressed this issue by determining the effects of blocking cAMP transport into the renal tubules with probenecid (0.1, 0.3 and 1 mM) on the increase in renal venous output of adenosine induced by adding cAMP (30 microM) to the perfusate of isolated rat kidneys. Addition of cAMP to the perfusate caused a marked increase in renal venous secretion of adenosine, an effect that was augmented, rather than inhibited, by probenecid. To test the hypothesis that the renal vasculature supports a cAMP-adenosine pathway, cultured rat preglomerular vascular smooth muscle cells were incubated with cAMP (30 microM) for 1 hr in the presence and absence of 3-isobutyl-1-methylxanthine (a phosphodiesterase inhibitor). Incubation with cAMP increased extracellular adenosine levels 41-fold, and this effect was abolished by 3-isobutyl-1 methylxanthine. In a third experimental series, addition of cAMP (0.3, 1, 3, 10 and 30 microM) to the perfusate of isolated rat kidneys and mesenteric vascular beds increased the renal venous, but not mesenteric venous, output of AMP, adenosine and inosine. We conclude that the renal vasculature supports a cAMP adenosine pathway, that administering cAMP into the renal artery and measuring adenosine in the venous effluent of the perfused rat kidney most likely monitors primarily the renal vascular cAMP-adenosine pathway and that the quantitative importance of the cAMP-adenosine pathway is not equivalent in all vascular compartments. PMID- 9336323 TI - Dose-dependent effects of the dopamine D1 receptor agonists A77636 or SKF81297 on spatial working memory in aged monkeys. AB - With advancing age, monkeys develop deficits in spatial working memory resembling those induced by lesions of the prefrontal cortex (PFC). Aged monkeys also exhibit marked loss of dopamine from the PFC, a transmitter known to be important for proper PFC cognitive function. Previous results suggest that D1 agonist treatment can improve spatial working memory abilities in aged monkeys. However, this research was limited by the use of drugs with either partial agonist actions or significant D2 receptor actions. In our study, the selective dopamine D1 receptor full agonists A77636 and SKF81297 were examined in aged monkeys for effects on the working memory functions of the PFC. Both compounds produced a significant, dose-related effect on delayed response performance without evidence of side effects: low doses improved performance although higher doses impaired or had no effect on performance. Both the improvement and impairment in performance were reversed by pretreatment with the D1 receptor antagonist, SCH23390. These findings are consistent with previous results demonstrating that there is a narrow range of D1 receptor stimulation for optimal PFC cognitive function, and suggest that very low doses of D1 receptor agonists may have cognitive-enhancing actions in the elderly. PMID- 9336324 TI - Effects of chronic caffeine administration on respiration and schedule-controlled behavior in rhesus monkeys. AB - The effects of chronic caffeine administration on ventilation and schedule controlled behavior were studied in 12 adult rhesus monkeys. In seated subjects prepared with a head plethysmograph, ventilation was measured during exposure to air (normocapnia) and to elevated levels of CO2 (3%, 4% and 5%) mixed in air (hypercapnia). Acute administration of caffeine (10.0-30.0 mg/kg i.m.) produced marked, dose-dependent increases in ventilation during conditions of normocapnia and hypercapnia. However, daily administration of caffeine (10.0 mg/kg i.m.) for 8 consecutive days resulted in tolerance to its respiratory-stimulant effects that was surmountable with higher doses. Caffeine-tolerant subjects also were cross-tolerant to theophylline, an active metabolite of caffeine, and to rolipram and Ro 20-1724, selective phosphodiesterase inhibitors. When chronic administration was terminated and the acute effects of caffeine were redetermined, sensitivity returned to levels obtained before chronic administration within 9 days. Drug effects on behavior were studied in monkeys trained to respond under a fixed-interval schedule of stimulus termination. Acute administration of caffeine (1.0-30.0 mg/kg i.m.) produced significant rate increasing effects on fixed-interval responding, but chronic administration resulted in tolerance that was insurmountable, such that no dose increased responding above control rates. Although the time course for development and loss of tolerance to the behavioral effects of caffeine corresponded closely with respiration, cross-tolerance did not extend to the behavioral effects of rolipram. Chronic caffeine administration had little effect on caffeine metabolism or clearance, which indicated that caffeine tolerance was pharmacodynamic. The results suggest that different neurochemical mechanisms mediate the effects of caffeine on respiration and behavior, and that inhibition of type IV phosphodiesterase plays a prominent role in caffeine-induced respiratory stimulation. PMID- 9336325 TI - Discriminative stimulus effects of magnesium chloride: substitution studies with monoamine uptake inhibitors and N-methyl-D-aspartate antagonists. AB - Previous studies suggest that magnesium chloride may have discriminative stimulus effects that partially overlap with those of noncompetitive N-methyl-D-aspartate antagonists as well as certain monoamine uptake inhibitors. In our study, rats were trained to discriminate 100 mg/kg magnesium chloride from saline and its discriminative stimulus effects were characterized with respect to N-methyl-D aspartate receptor and monoamine transporter functions in substitution tests. The discriminative stimulus effects of magnesium chloride were acquired within a moderate number of training sessions and showed dose-related substitution after either subcutaneous (3-300 mg/kg) or intracerebroventricular (0.3-300 microg) administration. The intracerebroventricular administration of magnesium chloride was over 4000 times more potent than its s.c. administration. The monoamine uptake inhibitors cocaine, GBR 12909, talsupram and citalopram fully substituted (> or =90% magnesium-appropriate responses) for magnesium chloride in the majority of subjects tested and the group averages reached a maximum of 72 to 82% responses on the magnesium-appropriate lever. Based on relative potency analysis, the rank order of potency of these four drugs for producing magnesium-appropriate responses was talsupram = cocaine > citalopram = GBR 12909. The N-methyl-D aspartate receptor antagonists dizocilpine, phencyclidine and NPC 17742 engendered maximum group averages of 49 to 65% responses on the magnesium appropriate lever. The results suggest that the centrally mediated discriminative stimulus effects of magnesium chloride may be more directly related to interactions with monoamine neurotransmitter functions than to N-methyl-D aspartate receptor blockade. PMID- 9336326 TI - The C-terminus of the thromboxane receptor contributes to coupling and desensitization in a mouse mesangial cell line. AB - To investigate regulatory domains of the thromboxane A2 (TxA2) receptor, we constructed a truncated form of the mouse TxA2 receptor and expressed it in a mesangial cell line. The mutant receptor lacked 22 amino acids in the C-terminus including four potential phosphorylation sites. Ligand binding of mutant receptors was identical with the wild type. Stimulation with TxA2 agonist induced increases in inositol trisphosphate (IP3) generation and [Ca++]i by both wild type and mutant receptors. However, the initial increase in IP3 generation by the mutant receptor was only approximately 50% of that seen in the wild type. Exposure of wild-type receptors to TxA2 agonist caused desensitization of IP3 and calcium responses. Pretreatment with TxA2 agonist caused some desensitization of mutant receptors, but the extent of desensitization was reduced compared with the wild type. The protein kinase C inhibitor staurosporine attenuated TxA2-induced desensitization of wild-type receptors, but had little effect on TxA2-induced desensitization of mutant receptors. Pretreatment with low concentrations of the phorbol ester, phorbol 12,13-dibutyrate (100 nM), reduced subsequent responsiveness of wild-type but not mutant TxA2 receptors. In contrast, high-dose phorbol 12,13-dibutyrate (1 microM) produced a similar degree of desensitization of both receptor types. These data suggest that: 1) the C-terminus participates in coupling of the TxA2 receptor to its effector systems; 2) the C-terminus contributes to agonist-specific desensitization of the TxA2 receptor; and 3) protein kinase C-induced desensitization of the TxA2 receptor is complex and depends, in part, on C-terminal domains of the TxA2 receptor. PMID- 9336327 TI - The pharmacological characterization of a novel selective 5-hydroxytryptamine1A receptor antagonist, NAD-299. AB - The pharmacological properties of a novel selective 5-hydroxytryptamine1A (5 HT1A) receptor antagonist, NAD-299 [(R)-3-N,N-dicyclobutylamino-8-fluoro-3,4 dihydro-2H-1-benzopyran-5-carboxamide hydrogen (2R,3R)-tartrate monohydrate] were examined in vitro and in vivo and compared with the reference 5-HT1A receptor antagonist, WAY-100635 [N-(2-(1-(4-(2-methoxyphenyl)piperazin-yl))ethyl)-N-(2 pyridinyl) cyclohexanecarboxamide trihydrochloride]. The new compound had high affinity for 5-HT1A receptors in vitro with a Ki value of 0.6 nM. The only other receptors for which NAD-299 had affinity less than 1 microM were alpha-1 and beta adrenoceptors with Ki values of 260 and 340 nM, respectively. Thus, the selectivity of NAD-299 for 5-HT1A receptors was more than 400 times. WAY-100635 had considerably higher affinity than NAD-299 for alpha-1 adrenoceptors (Ki = 45 nM) and dopamine D2 and D3 receptors (Ki = 79 and 67 nM, respectively). Like WAY 100635, NAD-299 competitively blocked 5-HT-induced inhibition of vasoactive intestinal peptide-stimulated cAMP production in GH4ZD10 cells and had no intrinsic activity. Both compounds were therefore 5-HT1A receptor antagonists in vitro and also behaved as such in in vivo experiments. Thus, they competitively antagonized the 8-hydroxy-2-(di-n-propylamino)tetralin-induced 5-HT behavioral effects, hypothermia, corticosterone secretion and inhibition of passive avoidance behavior without causing any actions of their own. The effective dose of NAD-299 varied between 0.03 and 0.35 micromol/kg s.c., depending on the test and the dose of 8-hydroxy-2-(di-n-propylamino)tetralin. PMID- 9336328 TI - Clozapine and haloperidol modulate N-methyl-D-aspartate- and non-N-methyl-D aspartate receptor-mediated neurotransmission in rat prefrontal cortical neurons in vitro. AB - The effects of the antipsychotic drugs haloperidol and clozapine on N-methyl-D aspartate (NMDA) and non-NMDA receptor-mediated neurotransmission were examined and compared in pyramidal cells of the medial prefrontal cortex in rat brain slices by using the techniques of intracellular recording and single-electrode voltage-clamp. The bath administration of either haloperidol or clozapine produced a marked facilitation (300-400%) of NMDA-evoked responses in a concentration-dependent manner. The EC50 values of haloperidol and clozapine were 38 and 14 nM, respectively. At concentrations of > or =100 nM, clozapine, but not haloperidol, produced bursts of excitatory postsynaptic potentials (EPSPs), which were blocked by glutamate receptor antagonists, suggesting that these EPSPs were the result of increasing release of excitatory amino acids. Haloperidol, but not clozapine, produced a concentration-dependent inhibition of alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid-induced current with an EC50 value of 37 nM. Haloperidol significantly decreased the amplitude of EPSPs evoked by the electrical stimulation of the forceps minor, whereas clozapine increased the amplitude of these EPSPs. The study of current-voltage relationship indicates that clozapine preferentially potentiates NMDA receptor-mediated transmission, whereas haloperidol depresses the non-NMDA receptor-mediated response, which probably obscures its potentiating effect on NMDA receptor-mediated EPSPs. PMID- 9336329 TI - ABT-089 [2-methyl-3-(2-(S)-pyrrolidinylmethoxy)pyridine]: I. A potent and selective cholinergic channel modulator with neuroprotective properties. AB - Accumulating preclinical and clinical evidence data suggests that compounds that selectively activate neuronal nicotinic acetylcholine receptor (nAChR) subtypes may have therapeutic utility for the treatment of several neurological disorders. In the present study, the in vitro pharmacological properties of the novel cholinergic channel modulator ABT-089 [2-methyl-3-(2-(S) pyrrolidinylmethoxy)pyridine], are described. In radioligand binding studies, ABT 089 was shown to display selectivity toward the high-affinity (-)-cytisine binding site present on the alpha4beta2 nAChR subtype (Ki = 16 nM) relative to the [125I]alpha-bungarotoxin binding site present on the alpha7 (Ki > or = 10,000 nM) and alpha1beta1deltagamma (Ki > 1000 nM) nAChR subtypes. In cation flux and channel current studies, ABT-089 displayed a more complex profile than (-) nicotine having agonist, partial agonist and inhibitory activities depending on the nAChR subtype with which it interacts. ABT-089 differentially stimulated neurotransmitter release. The compound displayed a similar potency and efficacy to (-)-nicotine to facilitate ACh release (ABT-089, EC50 = 3 microM; (-) nicotine, EC50 = 1 microM), but was markedly less potent and less efficacious than (-)-nicotine to stimulate dopamine release (ABT-089, EC50 = 1.1 microM; (-) nicotine, EC50 = 0.04 microM). Additionally, ABT-089 was neuroprotective against the excitotoxic insults elicited by exposure to glutamate in both rat cortical cell cultures (EC50 = 10 +/- 3 microM) and differentiated human IMR32 cells (EC50 = 3 +/- 2 microM). The differential full agonist/partial agonist profile of ABT 089, as compared with (-)-nicotine and ABT-418, illustrates the complexity of nAChR activation and the potential to target responses at subclasses of the neuronal and peripheral receptors. PMID- 9336330 TI - ABT-089 [2-methyl-3-(2-(S)-pyrrolidinylmethoxy)pyridine dihydrochloride]: II. A novel cholinergic channel modulator with effects on cognitive performance in rats and monkeys. AB - ABT-089 [2-methyl-3-(2-(S)-pyrrolidinylmethoxy)pyridine dihydrochloride], a novel ligand at neuronal nicotinic acetylcholine receptors with reduced adverse effects and improved oral bioavailability relative to (-)-nicotine, was tested in a variety of cognitive tests in rats and monkeys. Administered acutely, ABT-089 only marginally improved the spatial discrimination water maze performance of septal-lesioned rats. However, more robust improvement (45% error reduction on the last training day) was observed when ABT-089 was administered continuously via subcutaneous osmotic pumps (minimum effective dose: 1.3 micromol/kg/day). Continuous infusion of (-)-nicotine produced comparable improvement in the spatial discrimination water maze performance of septal-lesioned rats, but a 40 fold higher dose of (-)-nicotine was required (62 micromol/kg/day). Continuous infusion of ABT-089 to aged rats enhanced spatial learning in a standard Morris water maze, as indexed by spatial bias exhibited during a probe trial conducted after 4 days of training, but not when they were subsequently trained in a two platform spatial discrimination water maze. The compound induced a small impairment in young rats on the standard water maze, but not on the two-platform task. A probe trial conducted after additional training in the standard water maze revealed no age or drug effects. ABT-089 did not affect performance of either the aged or young rats during inhibitory (passive) avoidance training. Also, continuous infusion of ABT-089 did not affect responses to acoustic startle or prepulse inhibition of acoustic startle in young, aged or septal-lesioned rats and did not affect locomotor activity in either sham-lesioned or septal-lesioned rats. In monkeys, acute administration of ABT-089 modestly improved the delayed matching-to-sample performance of mature, adult monkeys and more robustly improved performance in aged monkeys. Improved performance in the aged monkeys was restricted to the longest delay intervals and was not accompanied by changes in response latencies. PMID- 9336331 TI - Chronopharmacological study of interferon-alpha in mice. AB - The influence of dosing time on the pharmacological effects (fever and antiviral activity) and the pharmacokinetics of interferon-alpha (IFN-alpha) was investigated in ICR male mice under light-dark (12:12) cycle. There was a significant circadian rhythm in rectal temperature, as an index of fever, at 0.5 hr after IFN-alpha (10.0 MIU/kg i.v.) injection. The rhythmic pattern resembled overall the rhythm that occurs in the nondrugged state. However, the percent change from basal level of rectal temperature varied according to the dosing time. The rhythmicity corresponded to the dosing time-dependent difference of PGE2 levels in thalamus after IFN-alpha injection, but it did not correspond to that of plasma IFN-alpha concentrations. A significant dosing time-dependent difference was also demonstrated for 2'-5'oligoadenylate synthetase activities, as an index of antiviral activity, in plasma and liver at 24 hr after IFN-alpha injection. It was related to the rhythmicity in plasma IFN-alpha concentrations that was caused by the rhythmicity in clearance of IFN-alpha. The choice of the most appropriate time of day for drug administration may help to achieve rational chronotherapeutics of IFN-alpha in certain experimental and clinical situations. PMID- 9336332 TI - Inactivation of nitric oxide synthase by substituted aminoguanidines and aminoisothioureas. AB - A series of substituted aminoguanidines and amino-substituted isothioureas have been examined as inhibitors of nitric oxide (NO) synthase (NOS) isoforms. Each of the agents produced a time- and concentration-dependent inactivation of the NO forming activity of the affinity-purified NOS isoforms. These inactivations required exposure of NOS to the drug under conditions that supported catalysis, consistent with the proposal that they act as alternate substrate, mechanism based inactivators. Of the aminoguanidines examined, 2-ethylaminoguanidine was the most efficient inactivator, exhibiting vs. iNOS an apparent KI value of 120 microM as measured at 100 microM arginine and a k(inact max) value of 0.48 min( 1) and thus an apparent second-order rate constant for inactivation of 4.0 mM( 1)min(-1). 2-Ethylaminoguanidine displayed a high isoform selectivity for the iNOS compared with the nNOS and eNOS isoforms. 2-Ethylaminoguanidine inactivated NO synthetic activity in cytokine-induced RAW 264.7 cells as measured at 100 microM extracellular arginine with an apparent KI value of 55 microM and a k(inact max) value of 0.09 min(-1). The inactivated RAW 264.7 cell NO synthetic capability was restored over a 3-hr period after drug removal to a value 60% of its pretreatment value. This recovery occurred despite the presence of cycloheximide sufficient to inhibit protein synthesis by >99%. 1-Amino-S methylisothiourea by contrast with the aminoguanidines was identified as a mechanism-based inactivator selective for the nNOS isoform. In contrast to S isopropylisothiourea, which was found to be both cell penetrant and reversible, 1 amino-S-methylisothiourea appeared cell impermeable and inhibited NOS enzyme "irreversibly." PMID- 9336333 TI - Inhibition of prostaglandin synthesis and effects of ethanol and pentobarbital in humans. AB - Results from animal research suggest that pretreatment with prostaglandin synthesis inhibitors (PGSIs) may inhibit physiological and behavioral effects of moderate ethanol ingestion. We examined the effects of ethanol and pentobarbital in humans with and without pretreatment with indomethacin, a potent PGSI. Ten male subjects with histories of recreational use of ethanol and sedative/hypnotics participated in this inpatient study. The effects of indomethacin alone (0.66 mg/kg), indomethacin (0, 0.17, 0.33, 0.66 and 1.33 mg/kg) in combination with ethanol (0 and 1 g/kg) and indomethacin (0 and 0.66 mg/kg) in combination with pentobarbital (0, 1.33 and 4 mg/kg) were tested. On test days, subjects swallowed capsules containing indomethacin or placebo. One hour later, they swallowed capsules that contained pentobarbital or placebo and a large drink (500 ml) of tonic water that contained ethanol or placebo (tonic water with 2 ml of ethanol floated on top). Both ethanol and pentobarbital affected subjective ratings, performance measures and heart rate. However, indomethacin pretreatment had no influence on drug-induced changes to ethanol and pentobarbital. The results of this study illustrate the relationship between depressant drugs and human performance, but they do not support the hypothesis that inhibition of prostaglandin synthesis diminishes the effects of ethanol and pentobarbital in humans. PMID- 9336334 TI - Interaction between hyperthermia and oxygen radical formation in the 5 hydroxytryptaminergic response to a single methamphetamine administration. AB - Administration of a single high dose of methamphetamine (METH) causes a rapid and reversible decrease in the activity of the tryptophan hydroxylase (TPH), the rate limiting enzyme in the synthesis of 5-hydroxytryptamine. This effect can be reversed completely by exposing the METH-impaired enzyme to a reducing environment, which suggests that the decrease in TPH activity is a reversible oxidative consequence of free radical formation. Consistent with this hypothesis, a single METH administration to male rats increased oxygen radical formation, as demonstrated by increased striatal dihydroxybenzoic acid formation after coadministration of salicylate with METH. Prevention of METH-induced hyperthermia attenuated both the increase in dihydroxybenzoic acid formation and the decrease in TPH activity observed 1 h after METH administration. These data suggest that both reactive oxygen species and hyperthermia contribute to the acute decrease in TPH activity which results from a single METH administration. PMID- 9336335 TI - Effect of cyclopiazonic acid on the force-frequency relationship in human nonfailing myocardium. AB - The present study investigated the functional role of the sarcoplasmic reticulum Ca++-ATPase in contraction and relaxation, intracellular Ca++-transients, as well as on the force-frequency relationship in human myocardium. The Ca++-ATPase activity of membrane vesicles isolated from sarcoplasmic reticulum (SR) obtained from nonfailing donor hearts (n = 7) was measured in the presence of cyclopiazonic acid (CPA, 0-30 microM), a highly specific inhibitor of the Ca++ ATPase of the SR (SERCA). The effects of CPA on parameters of contraction and relaxation, force-frequency relationship and [Ca++]i transients (with fura-2) were studied on isolated left ventricular muscle strips from human nonfailing myocardium. CPA concentration-dependently inhibited SERCA activity of isolated SR vesicles. In the presence of CPA (30 microM) the former positive force-frequency relationship in human left ventricular nonfailing myocardium became negative. Especially at high frequencies of stimulation, CPA decreased developed tension, peak rate of tension rise and systolic fura-2-light emission, whereas time to peak tension, time to peak [Ca++]i, time to 95% relaxation, diastolic tension and diastolic Ca++ levels were increased. Peak rate of tension decay and time to half relaxation and half-decay of [Ca++]i were not altered significantly after treatment with CPA. These findings provide evidence that the SERCA plays a functional role in the frequency-dependent increase in force of contraction in human myocardium. Because an impaired function of the SERCA is predominantly followed by alterations of inotropic and to a lesser degree of lusitropic function, other important factors to lower [Ca++]i and influence relaxation may be present in human myocardium to compensate for the reduced SERCA activity, e.g., Na+-Ca++ exchanger. PMID- 9336337 TI - Relationship between phospholipase D activation and endothelial vasomotor dysfunction in rabbit aorta. AB - Lysophosphatidylcholine (lysoPC) causes endothelial vasomotor dysfunction in isolated blood vessels, although the signaling pathways involved in this effect remain to be established. Although lysoPC stimulated phospholipase D (PLD) activity in cultured endothelial cells, the role of PLD in the vascular effects of lysoPC remains unclear. This study investigated the hypothesis that PLD is involved in lysoPC-induced endothelial vasomotor dysfunction in isolated rabbit aorta. LysoPC (3-30 microM) stimulated vascular PLD activity and inhibited endothelium-dependent vasorelaxation to acetylcholine within an identical concentration range. In contrast, lysoPC-induced inhibition of vasorelaxation was not prevented by the selective protein kinase C (PKC) inhibitor, GF109203X (3 microM), which suggested that this enzyme was not involved in the endothelial vasomotor dysfunction produced by lysoPC. The ability of two other lysophospholipids, lyso-platelet-activating factor (3-30 microM) and lysophosphatidylserine (10-30 microM) to induce endothelial vasomotor dysfunction was also associated closely with their ability to stimulate vascular PLD activity. Parallel stimulation of PLD activity and inhibition of acetylcholine induced relaxation was also observed with orthovanadate (0.1-3 mM), which suggested that the association between PLD activation and endothelial vasomotor dysfunction was not a phenomenon particular to lysophospholipids. The magnitude of PLD stimulation and the extent of endothelial dysfunction induced by these diverse stimuli were highly correlated (r2 = 0.88). These observations suggest that the PLD signal transduction pathway is important in the endothelial vasomotor dysfunction produced by lysophospholipids and perhaps other agents. PMID- 9336336 TI - Kinetic evidence for active efflux transport across the blood-brain barrier of quinolone antibiotics. AB - A distributed model has been used to clarify the mechanism of the restricted and differential distribution of the quinolone antibiotics in the rat central nervous system (CNS). The symmetrical permeability clearances across the blood-brain barrier (BBB), PS(BBB), and across the blood-cerebrospinal fluid barrier (BCSFB), PS(CSF), and the active efflux clearances across the BBB, PS(BBB,eff), were obtained from a nonlinear least squares regression analysis combined with the fast inverse Laplace transforming program for in vivo data. The values of PS(BBB,eff) were 10- to 260-fold greater than those of PS(BBB), providing kinetic evidence to support the hypothesis that a significant efflux transport across the BBB is responsible for the limited distribution of quinolones in brain tissue. Moreover, by simulation studies, we could demonstrate the concentration profiles in the brain as a function of the distance from the ependymal surface. However, active efflux transport across the BCSFB has been suggested to have only a slight effect on the apparent elimination from the cerebrospinal fluid. Comparing the apparent brain tissue-to-unbound serum concentration ratio at steady state, it has been suggested that the net flux across the BBB, ie., the ratio of PS(BBB) to the sum of PS(BBB) and PS(BBB,eff), is a determinant for the differential distribution of these quinolones in brain tissue. Such a putative active efflux transport system would play a significant role in decreasing the brain interstitial fluid concentration of quinolones. PMID- 9336338 TI - Chlorpyrifos produces selective learning deficits in rats working under a schedule of repeated acquisition and performance. AB - Chlorpyrifos (CPF) is a cholinesterase-inhibiting organophosphate pesticide used extensively to treat crops and domestic animals. Two experiments determined the effects of acute and repeated CPF exposure on the acquisition and performance of response sequences. Adult male Long-Evans rats (n = 16), maintained at 300 g body weight were trained using food reinforcement under a multiple schedule of repeated acquisition (RA) and performance (P). The RA component required completion of a four-response sequence on three levers (e.g., center, right, left, right) that changed with each session, while the correct sequence in the P component was invariant. In experiment I, rats were orally administered vehicle (corn oil), 12.5, 25, 37.5 and 50 mg/kg CPF. Doses of 37.5 and 50 mg/kg produced greater accuracy decreases in RA than in P, suggesting a selective learning deficit. In experiment II, the rats were divided into two groups (n = 7), and received either vehicle or 12.5 mg/kg CPF, 5 day/wk, for 8 wk. Although 12.5 mg/kg CPF was barely effective when administered acutely, when administered repeatedly it initially decreased accuracy in both RA and P. Tolerance developed to CPF effects on P accuracy but not on RA accuracy. Microanalyses of response patterns indicated the most common type of error was a progression through the sequence as if incorrect responses were actually correct. Radiometric analyses of serum cholinesterase activity showed CPF produced 90% inhibition at 3 hr and 85% inhibition at 24 hr postexposure. These results show that both acute and repeated CPF produced a selective deficit in the learning of response sequences in rats. This selectivity was most clearly expressed through the development of tolerance to the disruptive effects of repeated CPF on the performance but not the learning of response sequences. PMID- 9336339 TI - FK-506 and cyclosporin A potentiate the IgE antibody production by contact sensitization with hapten in mice. AB - Five repeated topical applications of 2,4-dinitrofluorobenzene to the ears of BALB/c mice resulted in contact dermatitis on the ears as well as significant elevation in dinitrophenol-specific IgE antibody and total IgE in the serum. FK 506 and cyclosporin A inhibited the development of contact dermatitis in terms of skin thickness and histopathological changes of skin lesions. On the contrary, these two drugs potentiated dinitrophenol-specific and total IgE antibody production without affecting IgG and IgM levels in serum. The expression of interferon-gamma mRNA in reverse transcriptase-polymerase chain reaction in the ear was inhibited by FK-506 and cyclosporin A. The expression of interleukin-4 mRNA, germline C epsilon and productive C epsilon in the auricular lymph node was not affected by these two drugs. Contrary to the above in vivo findings, the immunosuppressors, FK-506 and cyclosporin A, inhibited the production of interferon-gamma and interleukin-2 by cultured Th1 cells (1E10.H2 cells) and of interleukin-4 and -5 by Th2 cells (D10.G4.1 cells) in vitro. These results indicated that FK-506 and cyclosporin A selectively inhibited the Th1 cell mediated contact dermatitis and potentiated the Th2 cell-mediated IgE antibody production in vivo. This potentiation is probably due to the down-regulation of interferon-gamma production by Th1 cells after the treatment with these drugs. However, because FK-506 and cyclosporin A inhibited the production of cytokines by both Th1 and Th2 cells in vitro and these two immunosuppressors showed higher selectivity toward inhibiting Th1 cell-mediated reactions by limitations in vivo experiments. PMID- 9336340 TI - Prevention of amyloid-like deposition by a selective prolyl endopeptidase inhibitor, Y-29794, in senescence-accelerated mouse. AB - Our study was performed to assess the hypothesis that prolyl endopeptidase (PEP) would be functionally involved in the senescence-accelerated amyloid formation and that long-term inhibition of prolyl endopeptidase would suppress the progression of A beta-like deposition in the hippocampus of the senescence accelerated mouse (SAM). Granular structures of A beta-LI were observed in the hippocampus and around cerebral microvessels of the SAM after immunohistochemical staining with specific anti-A beta antibody. Repeated treatment of the SAM with Y 29794 (1, 10, 20 mg/kg, p.o.), a specific inhibitor of prolyl endopeptidase, significantly reduced the number and density of A beta-positive granular structures in the hippocampus of the SAM, after digital image analysis with NIH Image software. Furthermore, the characteristic biphasic distribution of the digitized density of the granules was significantly modulated after the treatment with Y-29794. These results suggest that chronic treatment of the SAM with Y 29794, a nonpeptide inhibitor of prolyl endopeptidase, prevents the progression of A beta-like deposition in the hippocampus of the SAM. PMID- 9336341 TI - Effects of the novel multiple-action agent carvedilol on severe nephrosclerosis in renal ablated rats. AB - Antihypertensive drugs have differing effects on renal hemodynamics and morphology. We analyzed whether the use of a new beta adrenoceptor antagonist and vasodilator, carvedilol (CVD), slows the progression of nephrosclerosis and whether the renoprotective effect as well as reduction in cardiac hypertrophy is dependent on the degree of blood pressure reduction. Fifty-four adult male Sprague-Dawley rats were distributed among five groups: group I served as untreated controls with 5/6 nephrectomy (Nx); group II, sham (no renal ablation or drug treatment); group III, CVD 5 (5/6 Nx and treatment with oral CVD at 5 mg/kg/day); group IV, CVD 10 (5/6 Nx and treatment with oral CVD at 10 mg/kg/day); and group V, CVD 20 (5/6 Nx and treatment with oral CVD at 20 mg/kg/day). Tail-cuff blood pressure and 24-hr urine samples were obtained before and at 3, 5 and 11 weeks of treatment with CVD. At the end of the study period, blood was taken to measure serum creatinine, plasma renin activity and CVD levels, as well as the remnant kidney and heart for morphological studies. There was a significant reduction in 24-hr U(ProtV) in all the CVD-treated groups, and it was increasingly evident with the highest dose used. However, only rats receiving doses of 10 and 20 mg/kg/day of CVD exhibited significant decreases in blood pressure. Elevated serum creatinine levels seen in untreated controls were significantly decreased by CVD in treated rats (P < .01), indicating that glomerular filtration rate was improved by this drug. This was associated with a significant increase in U(NaV). Concomitant and significant (P < .01) decreases in plasma renin activity were observed in sham and CVD-treated rats. CVD-treated animals had considerably reduced renal damage (P < .01) and cardiac hypertrophy (P < .01) compared with untreated controls. These data indicate that CVD is effective in delaying progression of renal damage and provides beneficial effects in the remnant kidney and cardiac hypertrophy, even at nonhypotensive doses. PMID- 9336343 TI - Pentobarbital decreases the gamma-aminobutyric acidA receptor subunit gamma-2 long/short mRNA ratio by a mechanism distinct from receptor occupation. AB - Treatment with pentobarbital of primary cultured cerebellar granule cells decreased the gamma-aminobutyric acid, (GABA)A receptor subunit gamma-2 long/short (gamma-2L/S) mRNA ratio. A high dose of pentobarbital (500 microM) decreased the gamma-2L/S ratio by 64%; the decrease was dose and time dependent and reversible. (-)-Hexobarbital (500 microM), the less potent stereoisomer for GABA(A) receptor activation, decreased the ratio slightly (30%) but significantly more than (+)-hexobarbital (20%). Other GABA(A) receptor activators had no (100 mM ethanol) or little (2 microM 5alpha-pregnane-3alpha-ol-20-one) effect on the gamma-2L/S ratio. Furthermore, picrotoxin (10 microM), which blocks the GABA- and pentobarbital-activated GABA(A) receptor channel, neither changed the gamma-2L/S ratio nor blocked the pentobarbital-induced changes. These data suggest that barbiturates alter the gamma-2L/S mRNA ratio by a mechanism that does not require GABA(A) receptor activation. The gamma-2L/S subunit mRNA includes an exon encoding an octapeptide that contains a protein kinase C phosphorylation consensus site. This exon-encoded peptide, occurring in the putative intracellular loop, can be phosphorylated, and in vitro, this phosphorylation has been shown to have functional consequences. This is the first report of a drug induced alteration in receptor mRNA splicing. Furthermore, the changes in the gamma-2L/S ratio produced by pentobarbital exposure may have significant effects on the function of an important brain protein, the GABA(A) receptor. PMID- 9336342 TI - Two differential effects of cyclic adenosine 3',5'-monophosphate on IL-5 production by antigen-specific human T cell line. AB - It has been proven that increasing cyclic adenosine 3',5'-monophosphate (cAMP) in human helper T cells results in decreased production of interleukin (IL)-2. As we have recently found that IL-2 stimulates IL-5 production, the effects of cAMP on IL-5 synthesis of T cells was investigated in this study. Prostaglandin E2 and forskolin raised intracellular cAMP level of Dermatophagoides farinae extract reactive human T cell line and inhibited T cell receptor-stimulated IL-5 production. The cAMP analog, dibutyryl-cAMP, also inhibited IL-5 production, whereas the protein kinase A inhibitor, H-89, enhanced IL-5 production. The IL-5 production was completely suppressed by anti-IL-2 neutralizing antibody. Recombinant human IL-2 itself induced IL-5 production, suggesting that IL-5 production stimulated through T cell receptor is dependent on the autocrine production of IL-2. Prostaglandin E2, forskolin and dibutyryl-cAMP enhanced but H 89 suppressed recombinant human IL-2-induced IL-5 production. Prostaglandin E2 suppressed T cell receptor-stimulated mRNA expression of IL-2 as well as IL-5 in the T cell line, whereas it potentiated IL-5 mRNA expression stimulated by recombinant human IL-2. These results suggest that the inhibitory effect of cAMP on IL-5 production is mediated by the suppression of IL-2 production. On the contrary, IL-2-induced IL-5 synthesis is enhanced by increasing cAMP. Our study clearly indicated that cAMP regulates IL-5 production of human T cells by two differential effects. PMID- 9336344 TI - An anti-inflammatory benzamide derivative inhibits the protein kinase C (PKC) dependent pathway of ERK2 phosphorylation in murine macrophages. AB - We have previously described benzamide derivatives that inhibited tumor necrosis factor (TNF) production from activated macrophages (Mphi) probably by interacting with a protein kinase C (PKC)-dependent pathway. To investigate their mode of action further, we first tested their effect on isolated PKC in vitro, using the selective inhibitor bisindolylmaleimide (BIM) as a positive control. We found that our representative compound JM34 did not inhibit PKC activity in vitro. We then investigated pathways located downstream of PKC and focused on the Raf1/MEK1,2/Erk1,2 cascade known to be preferentially activated by PKC activators such as phorbol esters. We found that JM34 dose-dependently inhibited Erk2 phosphorylation in Mphi stimulated by phorbol dibutyrate and calcium ionophore (maximal inhibition of 85% at 300 microM). BIM at 3 microM totally abrogated Erk2 phosphorylation. After stimulation with endotoxin or zymosan, Erk2 phosphorylation was only partially inhibited (25-30%) by JM34 or BIM, which confirmed that PKC-independent events were also involved in Erk2 phosphorylation. Because activated Erk2 has been shown to activate phospholipase A2, we tested the effect of JM34 and BIM on the release of arachidonate metabolites from activated Mphi. We found that both products partially inhibited the release of arachidonate metabolites from zymosan-activated Mphi at levels comparable to their inhibition of Erk2 phosphorylation. In contrast, JM34 and BIM markedly differed in their ability to inhibit TNF production. Taken together, our results suggest that JM34 inhibited the PKC-dependent pathway of Erk2 phosphorylation, which may fully account for its inhibitory effect on phospholipase A2 activation. However, the inhibition of TNF release by JM34 probably involved inhibition of an additional pathway, distinct from the Erk1/Erk2 cascade. PMID- 9336345 TI - Insulin-like growth factor (IGF) gene expression is reduced in neural tissues and liver from rats with non-insulin-dependent diabetes mellitus, and IGF treatment ameliorates diabetic neuropathy. AB - Neural disturbances are observed in the peripheral and central nervous systems of patients with insulin-dependent diabetes mellitus (IDDM) and non-IDDM (NIDDM). Insulin-like growth factors (IGFs) are neurotrophic growth factors that can support nerve regeneration and neuronal survival in the types of neurons known to be afflicted in diabetes. We tested the hypotheses that IGF gene expression is reduced in neural tissues and liver of spontaneously diabetic obese Zucker (fa/fa) rats and that IGF treatment can prevent neuropathy. There was a significant early reduction in IGF-II mRNA content as measured per mg of wet tissue or per poly(A)+ RNA in sciatic nerves, spinal cord and brain from spontaneously diabetic obese (fa/fa) vs. nondiabetic lean (+/+) adult rats. In addition, IGF-I mRNA content was reduced in liver but not nerve or spinal cord of NIDDM rats. Pain/pressure thresholds were abnormal (hyperalgesia) in diabetic (fa/fa) vs. nondiabetic (+/+) rats, and subcutaneous infusion of IGF-II restored thresholds toward normal. The low dose of IGF-II that prevented hyperalgesia in contrast had no effect on hyperglycemia or obesity. These data suggest that IGF treatment may provide rational therapy for diabetic neuropathy and that therapy may be effective even in patients unable to adequately control their hyperglycemia. PMID- 9336346 TI - The novel calcium sensitizer levosimendan activates the ATP-sensitive K+ channel in rat ventricular cells. AB - Levosimendan, a new Ca++-sensitizing and positive inotropic agent, was reported to act as a coronary vasodilator and protect ischemic myocardium. To elucidate the mechanisms of these actions, the possible electrophysiological effects of levosimendan on isolated rat ventricular cells were examined by the patch-clamp technique with whole-cell and single-channel recordings. Levosimendan (3 and 10 microM) markedly shortened action potential duration and activated an outward current at potentials positive to -70 mV. The increased current was abolished by glibenclamide, a blocker of the ATP-sensitive K+ (K[ATP]) current. Stimulation of K[ATP] current was dose dependent, with an EC50 value of 4.7 microM; a maximal effect occurred at 30 microM. The L-type Ca++ current was not affected by levosimendan (0.2-10 microM). In single-channel current recording in open cell attached patches, K[ATP] channels, which had been inhibited by 0.3 mM ATP, were activated by levosimendan. However, levosimendan did not stimulate the K[ATP] channels that exhibited high spontaneous activity in ATP-free solution. Levosimendan also could not stimulate K[ATP] channels that had rundown in ATP free solution. However, levosimendan could stimulate rundown K[ATP] channels that were reactivated by nucleotide diphosphates. K[ATP] channels inhibited by 0.5 mM AMP-PNP, a nonhydrolyzable ATP analog, were not stimulated by levosimendan; however, the channels were stimulated by levosimendan in the presence of 30 to 50 microM ADP. Levosimendan stimulates cardiac K[ATP] channels that are suppressed by intracellular ATP. It appears that levosimendan acts synergistically with nucleotide diphosphates. These properties of levosimendan may help protect ischemic myocardium because activation of K[ATP] channels by levosimendan would likely occur in ischemic regions in which intracellular ADP concentration is increased and intracellular ATP concentration is decreased. PMID- 9336348 TI - Kinetic analysis of the disposition of MRK16, an anti-P-glycoprotein monoclonal antibody, in tumors: comparison between in vitro and in vivo disposition. AB - The kinetics of the disposition of MRK16, an anti-P-glycoprotein monoclonal antibody, was studied in two human colorectal tumor cell lines, HCT-15 and COLO205, whose P-glycoprotein expression is extensive and poor, respectively. In a series of in vitro binding studies, the amount of MRK16 associated with HCT-15 cells at steady state was approximately 40 times greater than that associated with COLO205 cells. In in vivo studies, the disposition of MRK16 was determined in tumor-bearing mice after intravenous administration. The difference in the tumor-to-plasma concentration ratio between the two cell lines was only 2.3-fold at 72 hr after injection. To explain the large difference observed between the in vitro and in vivo results, a series of kinetic simulation studies were performed. By considering the physiological parameters specific for MRK16 (such as permeability-surface area product and the kinetic parameters determined in vitro), the time profiles for the tumor concentration were predicted. The predicted difference in the tumor-to-plasma concentration ratio at 72 hr was calculated to be 2.6-fold, although the permeability-surface area product across the tumor capillary and other physiological parameters were comparable between the two tumor cell lines. The discrepancy between the in vitro and in vivo results was accounted for by the fact that the tumor extracellular fluid concentration at this time point was 13-fold lower in HCT-15 tumors than in COLO205 tumors because of the restricted penetration of MRK16 through the tumor capillaries. This finding suggests that this factor accounts for the in vitro and in vivo difference in the tumor disposition of MRK16. PMID- 9336349 TI - Inhibition of cell growth: effects of the tyrosine kinase inhibitor CGP 53716. AB - The growth factors, platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) play major roles in enhanced smooth muscle cells growth in rodent blood vessels after vascular injury. Tyrosine kinase inhibition has been shown to be effective in blocking tyrosine phosphorylation at the PDGF and bFGF receptors in cultured fibroblast and vascular smooth muscle cells which in turn inhibits their proliferation. Our study evaluated the PDGF selective tyrosine kinase inhibitor, CGP 53716, on serum, PDGF-BB, bFGF or epidermal growth factor induced growth responses in cultured rat aortic smooth muscle cells (RASMC) and Balb/3T3 fibroblasts (3T3). CGP 53716 inhibited serum-induced cell growth in RASMC, but not in 3T3 cells. CGP 53716 completely blocked PDGF-BB tyrosine receptor autophosphorylation in RASMC and 3T3 cells, PDGF-BB-induced phosphorylation of mitogen-activated protein kinase at 1 microM in RASMC and inhibited PDGF-BB-induced c-Fos protein expression at 1 microM in RASMC; consistent with inhibition of PDGF-BB-induced DNA synthesis. To examine the selectivity of CGP 53716, PDGF-BB, bFGF or EGF-induced DNA synthesis was measured using thymidine incorporation. CGP 53716 inhibited PDGF-BB-, bFGF- and EGF induced DNA synthesis in a concentration-dependent manner in each cell line. CGP 53716 showed a 2- to 4-fold selectivity for PDGF-BB-stimulated DNA synthesis over bFGF or EGF in RASMC or 3T3 cells. To rule out that bFGF induced the release of endogenous PDGF, an antibody to PDGF-AB, which binds to all three isoforms of PDGF, was coincubated with bFGF and did not suppress the DNA synthesis induced by bFGF. Based on these results, CGP 53716 is not selective for the PDGF receptor as previously reported. However, EGF-stimulated receptor autophosphorylation of mitogen-activated protein kinase phosphorylation and c-Fos protein expression were not inhibited by CGP 53716 at 1 or 10 microM in RASMC. These findings suggest that CGP 53716 may inhibit multiple growth factor pathways as indicated by inhibition of DNA synthesis. However, these effects must be downstream from the signaling for c-Fos protein expression or use an alternate signaling route. These results further suggest that CGP 53716 may have a therapeutic potential for the treatment of vascular proliferative diseases which are stimulated by not only PDGF but other growth factors such as bFGF and EGF. PMID- 9336347 TI - Sequestration of gamma-aminobutyric acidA receptors on clathrin-coated vesicles during chronic benzodiazepine administration in vivo. AB - Chronic administration of benzodiazepine agonists produces behavioral tolerance. For induction of tolerance, the use-dependent down-regulation of gamma aminobutyric acidA (GABA[A])/ benzodiazepine receptors is a potential cellular mechanism. We previously identified GABA(A) receptors on clathrin-coated vesicles from rat brain, suggesting that surface receptors can be internalized via endocytosis. To examine a role for coated vesicles in GABA(A) receptor down regulation in vivo, fractions were obtained from mouse brain microsomes through density centrifugation and treatment with 0.1% Triton X-100. This coated vesicle preparation was enriched in clathrin subunits and clathrin light-chain kinase and had twice the level of [3H]flunitrazepam binding as did vesicles not exposed to Triton. Adult mice were treated with lorazepam (2 mg/kg/day) for 7 days via osmotic minipump, achieving a serum level of 103 +/- 8.9 ng/ml. The level of flunitrazepam bound to coated vesicles was increased by 83 +/- 13% in the lorazepam-treated mice compared with vehicle-treated controls. The Bmax value for [3H]flunitrazepam binding to synaptic membranes from lorazepam-treated animals was 33 +/- 4% lower than that of controls. The amount of GABA(A) receptor alpha-1 subunits, as quantified by Western blotting, followed a similar pattern. Relative to controls, immunoreactivity for alpha-1 subunits in coated vesicles from lorazepam-treated mice was increased by 60.0 +/- 10.3%, whereas that in synaptic membranes declined by 12 +/- 6%. These results indicate that lorazepam-dependent GABA(A) receptor sequestration occurs in mouse brain. Furthermore, it is suggested that this sequestration may play a role in GABA(A) receptor down regulation in vivo. PMID- 9336350 TI - Characterization of functional chemokine receptors (CCR1 and CCR2) on EoL-3 cells: a model system to examine the role of chemokines in cell function. AB - A growing family of proteins, known as the chemokines, play an important role in the recruitment and activation of inflammatory cells. The purpose of these studies was to characterize the chemokine receptors present on human sodium butyrate differentiated EoL-3 cells (dEoL-3 cells). Using a combination of 3' rapid amplification of cDNA ends and nested polymerase chain reaction, we detected mRNA for CC chemokine receptor (CCR)1, CCR2, CCR3 and low level of CCR5. Radioligand binding studies demonstrated high-affinity saturable binding for both 125I-macrophage inflammatory protein (MIP)-1alpha and 125I-regulated upon activation normal T cell expressed and secreted (RANTES) with Kd values of 1.4 and 7 nM, respectively. Competition binding with chemokines demonstrated exactly the same rank order of potency for displacement of both ligands: MIP-1alpha approximately monocyte chemoattractant protein (MCP)-3 approximately RANTES > MIP 1beta >> MCP-1 >>> IL-8. RANTES, MCP-3 and MIP-1alpha all produced concentration dependent transient increases in intracellular calcium concentrations in dEoL-3 cells. Desensitization studies indicated that RANTES, MIP-1alpha and MCP-3 interacted at the same receptor, which is identical in characterization to the cloned CCR1. 125I-MCP-1 also demonstrated high-affinity satuable binding to dEoL 3 cells with a Kd value of 0.4 nM. Competition studies showed that MCP-3 was slightly more potent than MCP-1 and MCP-2. MIP-1alpha, MIP-1beta and RANTES were unable to displace 125I-MCP-1. Addition of either MCP-1 or MCP-3 produced a concentration-dependent elevation of intracellular calcium with a maximun response 2-fold higher than that seen with RANTES or MIP-1alpha. Desensitization studies indicated that MCP-1 and MCP-3 function through CCR2 on these cells. Thus binding and functional studies indicate that dEoL-3 cells express functional CCR1 and CCR2 and that these cells may serve as an important system with which to study the regulation and role of these receptors. PMID- 9336351 TI - Selection bias in trials of transplantation for metastatic breast cancer: have we picked the apple before it was ripe? PMID- 9336352 TI - Impact of selection process on response rate and long-term survival of potential high-dose chemotherapy candidates treated with standard-dose doxorubicin containing chemotherapy in patients with metastatic breast cancer. AB - PURPOSE: Most of the data about high-dose chemotherapy (HDCT) for metastatic breast cancer are derived from phase II studies. The interpretation of these data depends on comparisons with data from properly selected historical control patients treated with standard therapy under similar circumstances. We report the long-term results of patients with metastatic breast cancer who were eligible for HDCT but were treated with doxorubicin-containing standard-dose chemotherapy. PATIENTS AND METHODS: Prospectively collected data from 18 successive doxorubicin containing protocols for the treatment of metastatic breast cancer were evaluated. Using common eligibility criteria for HDCT, we identified patients who would have been candidates for HDCT. We analyzed response rates, progression-free survival (PFS), and overall survival (OS) for all patients, potential HDCT candidates, and noncandidates. RESULTS: A total of 1,581 patients was enrolled onto the 18 studies. Six hundred forty-five were HDCT candidates, and 936 were noncandidates. The complete response rate was 27% for HDCT candidates and 7% for noncandidates; median PFS was 16 and 8 months and median OS was 30 and 17 months, respectively. Survival rates for HDCT candidates and noncandidates, respectively, were 21% and 6% at 5 years and 7% and 2% at 10 years. CONCLUSION: This study suggests that encouraging results of single-arm trials of HDCT could partially be due to selection of patients with better prognoses and further stresses the importance of completing ongoing randomized trials of HDCT to assess the relative efficacy of HDCT in patients with metastatic breast cancer. PMID- 9336353 TI - Patient selection in high-dose chemotherapy trials: relevance in high-risk breast cancer. AB - PURPOSE: To evaluate the impact of the selection criteria that are used in current high-dose consolidation chemotherapy (HDCT) trials on the outcome of high risk breast cancer patients treated with conventional adjuvant chemotherapy. PATIENTS AND METHODS: From 1975 to 1995, 265 breast cancer patients at our institution showed involvement of ten or more positive axillary lymph nodes. Of these, 171 received standard adjuvant combination chemotherapy, but not HDCT consolidation, and were the subjects of our study. One hundred twenty-eight patients met the standard selection criteria for HDCT with hematological support (< 60 years, no significant concomitant disease, and no progression during adjuvant treatment), whereas 43 did not. Clinical outcome was analyzed by using disease-free survival (DFS) and overall survival (OS) as endpoints. To provide an assessment of the short-term efficacy of HDCT, we also evaluated the outcome in a cohort of 39 patients from the last 4 years who met the criteria for, and were actually treated with, HDCT after adjuvant chemotherapy. RESULTS: With a median follow-up of 4.4 years (range, 0.7 to 17.2 years), 112 of the 171 patients have had a relapse, and 87 have died. The estimated 5-year DFS was 32.3%, and OS was 49.4%. DFS was significantly higher for patients who met the HDCT criteria (36.6% at 5 years) than for those who did not (15.8% at 5 years; P < .05). OS was also significantly more favorable in patients meeting HDCT criteria (55.4% at 5 years) than in patients not meeting HDCT criteria (22.7% at 5 years; P < .01). We performed a multivariate analysis to adjust for other potential prognostic factors and found that meeting the HDCT criteria and having undergone locoregional radiotherapy were the only significant independent predictors for DFS and OS. Finally, we compared the outcome of the 128 patients who met the HDCT criteria and were treated with conventional adjuvant chemotherapy only with that of the 39 patients who met the criteria and who actually underwent HDCT, and we did not observe significant differences in the DFS or OS between these groups. CONCLUSIONS: Meeting HDCT inclusion criteria is an independent indicator of favorable prognosis in high-risk breast cancer patients. The selection of patients by these criteria may explain, at least in part, the promising short term results of nonrandomized adjuvant HDCT trials in high-risk breast cancer. PMID- 9336354 TI - Treatment of advanced breast cancer with sterically stabilized liposomal doxorubicin: results of a multicenter phase II trial. AB - PURPOSE: A multicenter phase II study to determine the activity and toxicity of Caelyx (Doxil; Sequus Pharmaceuticals Inc, Menlo Park, CA) in patients with metastatic breast cancer. PATIENTS AND METHODS: Seventy-one patients with stage IV breast cancer were treated with Caelyx at doses of 45 to 60 mg/m2 every 3 to 4 weeks for a maximum of six cycles. Twenty-eight patients had received prior chemotherapy with a nonanthracycline regimen. Fifty-two patients had disease at multiple sites. Hepatic and pulmonary disease were the predominant metastatic site in 50 patients. Response was assessable in 64 cases. RESULTS: Sixteen patients achieved a partial response and a complete response (overall response rate, 31%; (95% confidence interval, 20% to 43%). Twenty patients (31%) had stable disease on treatment. Neutropenia > or = grade 3 occurred in 10% of cycles (27% of patients) and mucositis > or = grade 3 in 10% of cycles (32% of patients). Significant alopecia was rare and routine prophylactic antiemetics were not required. At doses of 60 mg/m2 every 3 weeks, seven of 13 patients had > or = grade 3 skin toxicity; overall, this toxicity complicated 25% of treatment cycles. The incidence of > or = grade 3 skin toxicity was greatly reduced at doses of 45 mg/m2 every 4 weeks, occurring in five of 32 patients and affecting only 5% of 126 treatment cycles. CONCLUSION: Caelyx is an active agent in advanced breast cancer with a safety profile that differs markedly from nonliposomal doxorubicin. A regimen of 45 mg/m2 every 4 weeks was well tolerated in this cohort of women with advanced poor-prognosis breast cancer. The mild myelosuppression seen with this regimen would favor its use in combination chemotherapy. PMID- 9336355 TI - Breast cancer patients' attitudes about rationing postlumpectomy radiation therapy: applicability of trade-off methods to policy-making. AB - PURPOSE: Along with evidence, clinical policies must take patients' values into account. Particularly where evidence is limited and where assumptions of utility maximizing behavior may not be valid, new methods such as trade-off techniques (TOTs), which allow elicitation of patients' treatment alternatives, might be useful in policy formulation. We used TOTs to assess breast cancer patients' attitudes toward two clinical policies designed to ration adjuvant postlumpectomy breast radiation therapy. METHODS: Cross-sectional interviews were performed in a tertiary cancer center. A total of 102 patients were presented with information about the side effects and benefits associated with two hypothetical decisions: (1) willingness to receive treatment elsewhere to shorten the wait for radiation therapy, and (2) foregoing radiation therapy in the face of small marginal benefits. For each scenario, a TOT was used to identify the maximal acceptable wait time (MAWT) for therapy and the benefit threshold at which the patient would forego therapy. Associations of clinical and demographic factors with these decisions were determined by regression analysis. RESULTS: Patients would be willing to wait, on average, 7 weeks before wanting to leave their city for radiation therapy, less than the 13-week delay our patients actually faced. Older patients were less willing to wait (P = .013); 46% of patients would not give up radiation therapy, even in the face of no stated benefit. Willingness to give up radiation therapy was predicted by willingness to accept delay (odds ratio [OR], 1.84; 95% confidence interval [CI], 1.05 to 3.37) and being employed (OR, 2.61; 95% CI, 1.08 to 6.54). Patients with larger tumors were less willing to give up radiation therapy (OR, 0.57; 95% CI, 0.31 to 0.97). CONCLUSION: Even in difficult decisions such as rationing postlumpectomy breast cancer radiation therapy, TOTs can inform policy formulation by indicating the distributions of patients' preferences. PMID- 9336356 TI - Effect of estrogen and estrogen-progestin replacement regimens on mammographic breast parenchymal density. AB - PURPOSE: Hormone replacement therapy (HRT) may increase the mammographic density with a possible reduction in the sensitivity or specificity. If so, the benefit of mammographic screening in women using HRT could be compromised. We evaluated the hypothesis that HRT regimens have differential effects on the mammographic density depending on treatment regimens or on age. PATIENTS AND METHODS: Among 31,498 Swedish women who received mammographic screening, we selected 554 women who started HRT after the first examination and who were current users at the second, and 554 age-matched women who had never received HRT. Mammograms were examined in a blinded review. The changes in density between the two examinations, graded as moderate or weak reduction, no change, or weak, moderate, or substantial increase, were assessed. We studied four HRT regimens-estradiol compounds only, estradiol compounds cyclically or continuously combined with progestins, and weak estrogens-and used descriptive statistics and logistic regression to analyze the association between HRT and density change. RESULTS: Density increased in 10% and 28% of women who received estradiol compounds with cyclically or continuously combined progestins, respectively, but in only 3% of unexposed women. Logistic regression analyses showed an elevated risk of a density increase (relative risk [RR] = 3.6; 95% confidence interval [CI], 1.6 to 7.7) in women who received cyclically combined regimens or continuously combined regimens (RR = 12.4; 95% CI, 6.3 to 24.4) compared with unexposed women. Women > or = 50 years of age had even stronger associations; RRs in women on estradiol only, the cyclically combined and the continuously combined regimens were 32.2 (95% CI, 3.9 to 267.5), 21.9 (95% CI, 1.9 to 251.5), and 176.9 (95% CI, 22.8 to 1,372.7), respectively. CONCLUSION: HRT with estradiol-progestin regimens, especially continuously combined, may increase the mammographic density in a substantial proportion of women. PMID- 9336357 TI - Revertant and potentiating activity of lonidamine in patients with ovarian cancer previously treated with platinum. AB - PURPOSE: Lonidamine (LND) is an energolytic derivative of indazol-carboxylic acid that has been shown to enhance cisplatin (CDDP) activity in both sensitive (A2780) and resistant (A2780/Cp8) ovarian cancer cell lines. The aim of this study was to confirm the potentiating or reverting activity of LND on CDDP activity obtained in experimental models in a phase II study of advanced ovarian cancer patients previously treated with platinum-based regimens. PATIENTS AND METHODS: Twenty-seven consecutive women with histologically proven and measurable ovarian cancer previously treated with platinum compounds were treated with CDDP plus LND. CDDP was administered at 1 mg/kg intravenously (IV) once weekly for 6 weeks and every 3 weeks thereafter until disease progression or toxicity. LND was administered at 450 mg daily (1 tablet every 8 hours) for the entire period of therapy starting 3 days before the first CDDP administration. In addition, a higher LND dosage was provided on the day of CDDP administration in an attempt to maximize the synergy of this drug with CDDP. RESULTS: Ten patients achieved a complete response (CR) or partial response (PR) for an overall response rate of 37% (95% confidence interval [CI], 19% to 55%). In particular, responses were observed in five of 18 (28%) refractory or early relapsed patients (one CR and four PRs) and in five of nine patients (55%) in the late-relapsed group (two CRs and three PRs). Grade 3 or 4 anemia, leukopenia, and thrombocytopenia were observed in 19%, 15%, and 11% of patients, respectively, whereas seven of 27 patients (26%) showed LND-related myalgia. Grade 3 renal toxicity was observed in two patients (8%). Neurotoxicity, often concealed by LND-related myalgia, was recorded as grade 1 or 2 in six patients (22%) and as grade 3 in one (4%). CONCLUSION: The 37% response rate observed in this study (28% in refractory or early-relapsed patients), suggests that the synergism between CDDP and LND observed in vitro against ovarian cancer cell lines can be clinically confirmed. However, larger series and randomized studies are needed to assess definitely the revertant activity of LND on CDDP-refractory patients. PMID- 9336358 TI - Fifteen-year survival and recurrence rates after radiotherapy for localized prostate cancer. AB - PURPOSE: To determine 15-year survival and recurrence rates after radiotherapy for localized prostate cancer. METHODS: One hundred thirty-six patients with clinically localized prostate cancer treated from 1966 to 1974 with interstitial gold seed and external-beam irradiation were evaluated to determine the probability of recurrence and survival > or = 15 years after therapy. All patients were surgically staged with pelvic lymphadenectomy and none received hormonal therapy before relapse. RESULTS: Overall, 60 patients (44%) have never recurred, although 57% (34 of 60) of these same patients have died of causes other than prostate cancer. Local progression developed in 39% of patients and distant metastases in 42%. At 15 years, the probability of dying of prostate cancer was 33%+/-8% (% +/- 2SE) and of all causes was 72%+/-8%. In clinical stage A2 and B, 29%+/-9% of patients died of their cancer within 15 years, compared with 57%+/-21% in stage C1, while only 18%+/-8% with clinical stage A2 and B and negative lymph nodes died of cancer within this period. In contrast, the prostate cancer mortality rate at 15 years was high for patients with positive nodes regardless of the stage of the primary tumor (73% for A2 and B; 71% for C1). Patients with nodal metastases, poorly differentiated tumors, and advanced local disease all had a significantly (P < .0001) increased risk of cancer death. CONCLUSION: The cancer-specific mortality rate for patients with stage A2 and B tumors and negative nodes compares favorably with other series of patients treated with radiation therapy and > or = 15 years' follow-up evaluation. While local progression rates are high and associated with a substantial risk of prostate cancer death, many patients live with the disease and ultimately die of causes other than prostate cancer. PMID- 9336359 TI - Quantitation of intratumoral thymidylate synthase expression predicts for disseminated colorectal cancer response and resistance to protracted-infusion fluorouracil and weekly leucovorin. AB - PURPOSE: Response rates to fluorouracil (5-FU)-based therapy remain low. As new, active agents are being tested, information regarding specific intratumoral genetic determinants of chemotherapy sensitivity or resistance can be used to plan therapy rationally. Intratumoral thymidylate synthase (TS) quantitation may be among the most important determinants of sensitivity or resistance to 5-FU. MATERIALS AND METHODS: Forty-six disseminated colorectal cancer patients had measurable tumor biopsies for polymerase chain reaction (PCR)-based determination of TS mRNA pretreatment. Protracted infusion of 5-FU 200 mg/m2/d for 21 days with weekly intravenous leucovorin 20 mg/m2 each cycle was given. After two cycles, responses were evaluated. Response data were correlated with independently determined intratumoral ratios of TS/beta-actin mRNA for each patient. RESULTS: TS/beta-actin ratios were successfully obtained for 42 patients (91%). TS/beta actin ratios ranged from 0.3 x 10(-3) to 18.2 x 10(-3) (median, 3.5 x 10[-3]). Twelve patients (26%) responded to treatment (median TS/beta-actin ratio, 1.7 x 10[+3]). Thirty-four patients did not respond (median TS/beta-actin ratio, 5.6 x 10[-3]). No patient with a TS mRNA level greater than 4.1 x 10(-3) responded. The median TS/beta-actin ratio (3.5 x 10[-3]) significantly segregated responders from nonresponders (P = .001). Median survival for patients with TS/beta-actin ratios < or = 3.5 x 10(-3) was 13.6 months; for patients with TS/beta-actin ratios greater than 3.5 x 10(-3), it was 8.2 months (P = .02). CONCLUSION: For this cohort, the intratumoral TS/beta-actin ratio had a statistically significant association with response and survival. This relationship for other 5-FU schedules remains unknown. Confirmation of these data in a larger patient population could lead to determination of therapy for disseminated colorectal cancer based on a specific intratumoral molecular parameter. PMID- 9336361 TI - Conservative treatment of rectal adenocarcinoma with endocavitary irradiation or wide local excision and postoperative irradiation. AB - PURPOSE: To evaluate the role of endocavitary irradiation and wide local excision followed by irradiation in the treatment of early-stage rectal adenocarcinoma. MATERIALS AND METHODS: Sixty-five patients with early-stage adenocarcinoma of the rectum were treated with endocavitary irradiation (n = 20) or wide local excision followed by external-beam irradiation (n = 45) between 1974 and 1994 at the University of Florida. All patients were monitored for a minimum of 2 years or until death. RESULTS: The rates of local-regional control at 5 years were 80% after endocavitary irradiation and 86% after wide local excision and radiotherapy. The ultimate 5-year local-regional control rates were 85% and 92%, respectively. Multivariate analysis of local-regional control with sphincter preservation showed that tumor configuration (exophytic v ulcerative) significantly influenced this end point; local-regional control was decreased in patients with ulcerated cancers. Five-year cause-specific survival rates were 84% after endocavitary irradiation and 88% after wide local excision and radiotherapy. Multivariate analysis revealed that tumor configuration significantly influenced cause-specific survival; patients with ulcerated tumors had a worse prognosis. CONCLUSION: Endocavitary irradiation is a highly effective treatment for properly selected patients with early-stage rectal adenocarcinoma. Patients with less favorable lesions that appear to be limited to the muscularis propria have a high chance of cure with sphincter preservation after wide local excision and external-beam irradiation. PMID- 9336360 TI - Assessment of genomic damage in colorectal cancer by DNA fingerprinting: prognostic applications. AB - PURPOSE: Here we evaluate the prognostic significance of the relative value of genomic damage assessed by DNA fingerprinting in colorectal cancer. MATERIALS AND METHODS: Sixty-three tumor and paired normal mucosa samples were included in the study. Genomic damage was assessed by comparative analysis of paired normal and tumor tissue DNA fingerprints by the arbitrarily primed polymerase chain reaction (AP-PCR). Decreases and increases of intensity in bands were computed and referred to the total number of visualized bands per case. An index reflecting the genomic damage fraction (GDF), with separated values for losses and gains, was obtained for each tumor. This index was used to determine molecular and clinicopathologic correlates after exclusion of eight cases displaying microsatellite instability. RESULTS: Fifty-five cases were considered for the statistical analysis. The average fraction of altered bands per tumor was 0.287+/ 0.121. When losses and gains were computed separately, the average fraction of changes was 0.126+/-0.113 and 0.161+/-0.120, respectively. Tumors lacking a ras mutation showed an increased GDF, primarily because of a higher fraction of gains. Tumors that were at advanced Dukes' stages and that were poorly differentiated also displayed a higher GDF. Finally, disease-free survival was significantly diminished in tumors with a GDF greater than 0.314 (P < .001). The prognostic significance of the GDF was independent of Dukes' stage (Cox multivariate analysis, P = .005). CONCLUSION: The degree of genomic damage assessed by unbiased DNA fingerprinting correlates with genotypic, phenotypic, and clinical variables in colorectal carcinoma and may be useful in assessing prognosis in colorectal cancer. PMID- 9336362 TI - Evaluation of newer prognostic markers for adult soft tissue sarcomas. AB - PURPOSE: In addition to tumor size, grade, location, and the presence of metastases, other factors may be useful in prognostication for adults with soft tissue sarcoma (STS). This study examines the relationship of MDR-1 mRNA, p glycoprotein (P-gp), Ki-67 expression, and DNA content expression to clinical outcome in adults with STS. PATIENTS AND METHODS: Snap-frozen STS specimens from 65 patients were analyzed and compared with clinical outcomes. Immunohistochemistry was performed for the Ki-67 antigen and P-gp. DNA content was determined using the Feulgen reaction and quantitated using image analysis. MDR-1 mRNA expression was determined using a reverse-transcriptase polymerase chain reaction (RT-PCR)-based assay. RESULTS: P-glycoprotein expression was found by immunohistochemistry in 48% of cases with 5-year overall (54% v 14%, P = .07) and disease-free survival rates (32% v 18%, P = .039) higher in high-grade tumors that did not express P-gp. MDR-1 mRNA was detected in 51% of cases and no patient with high levels of MDR-1 mRNA expression was a long-term survivor. Patients with diploid tumors had significantly better survival than those with nondiploid tumors (51% v 31%, P = .03). High levels of Ki-67 were associated with poorer overall survival (46% v 31%, P = .04). On multivariate analysis, American Joint Committee on Cancer (AJCC) staging, DNA content, Ki-67, and P-gp staining were significant prognostic factors for 5-year overall and disease-free survival. CONCLUSION: P-gp expression, high-level Ki-67 expression, and nondiploid DNA content are independent prognostic indicators that correlate with poor outcomes in STS patients. However, MDR-1 mRNA was not found to be predictive of survival. These newer markers are useful additions to AJCC staging for prognostication for patients with STS. Such markers may be useful in selecting high-risk STS patients who could benefit from systemic adjuvant therapy. PMID- 9336363 TI - High-dose cyclophosphamide for poor-prognosis and recurrent pediatric brain tumors: a dose-escalation study. AB - PURPOSE: To determine the maximum-tolerated dose (MTD) of cyclophosphamide (CTX) when administered over 2 consecutive days followed by hematopoetic stem-cell rescue given as two sequential courses to children with glioblastoma multiforme, poor-prognosis pontine gliomas, and other recurrent CNS tumors. PATIENTS AND METHODS: Two identical doses of CTX were administered 24 hours apart to 14 children and followed by hematopoetic stem-cell rescue. This treatment was repeated immediately following hematologic recovery. The starting dose of CTX was 2.5 g/m2/d with increments of 0.5 g/m2/d. CTX pharmacokinetics and metabolism were measured during 22 courses of treatment. Toxicity and tumor response were recorded. RESULTS: There were two toxic deaths at the dose level of 4 g/m2/d. These were not clearly related to cardiac toxicity and may have been due to generalized capillary leak syndrome. Thus, the MTD of CTX was 3.5 g/m2/ d. There were six complete responses (CRs) (46%; (95% confidence interval [CI], 19% to 73%) and four partial responses (PRs) (31%; 95% CI, 6% to 56%), and one patient achieved stable disease. All children with intracranial primitive neuroectodermal tumors (PNETs) improved following CTX. The median duration of tumor response was 6 months (range, 4 to 29) and only one patient remains disease-free following CTX alone. Overall survival is 21% (95% CI, 13% to 29%) at a median follow-up time of 27 months (range, 12 to 34). CONCLUSION: The MTD of CTX when followed by hematopoetic stem-cell rescue is 3.5 g/m2 administered on each of 2 consecutive days. This treatment was tolerable in children with poor-prognosis brain tumors and produced complete responses in children with recurrent PNETs. PMID- 9336365 TI - Phase II trial of docetaxel in non-Hodgkin's lymphomas: a study of the Cancer and Leukemia Group B. AB - PURPOSE: To evaluate the new anticancer agent, docetaxel, with a novel mechanism of action in patients with non-Hodgkin's lymphoma International Working Formulation (IWF) A through H, to determine the response rate by histologic group and the toxicities of this agent in this population. PATIENTS AND METHODS: Sixty eight patients previously treated for non-Hodgkin's lymphoma with two prior cytotoxic regimens for low-grade and one prior regimen for intermediate-grade lymphoma were entered onto this phase II trial. Central pathologic review was required. Twenty-four IWF A to C and 31 IWF D to H patients with normal hepatic and renal function, performance status (PS) 0 to 2, and adequate hematologic function were eligible. Patients received docetaxel 100 mg/m2 intravenously over 1 hour without corticosteroid premedications every 3 weeks with weekly hematologic monitoring, and tumor assessment every 3 weeks. For grade 3 or 4 hematologic toxicity, the docetaxel dosage was lowered to 75 mg/m2. Patients received a maximum of six cycles of therapy. RESULTS: The major response rate was 13% (95% confidence limits, 3% to 32%) for IWF A to C and 16% (95% confidence limits, 5% to 34%) for IWF D to H; response durations ranged from 1.4 to 20 months. Time to response ranged from 1.3 to 2.8 months. Patients refractory to previous chemotherapy were less apt to respond to docetaxel, but the differences were not statistically different in this small sample size. Twelve percent of IWF A to C and 6% of IWF D to H patients discontinued treatment because of toxicity. The major toxicity was granulocytopenia (grade 3 to 4), which occurred in virtually all patients during the first course of therapy. CONCLUSION: This study confirms that docetaxel has limited but definite activity in patients with non Hodgkin's lymphoma and suggests that the previously reported responses with taxanes can not be attributed solely to the use of corticosteroid premedications. PMID- 9336366 TI - Karnofsky Memorial Lecture. Hereditary cancer: theme and variations. PMID- 9336364 TI - IDEC-C2B8: results of a phase I multiple-dose trial in patients with relapsed non Hodgkin's lymphoma. AB - PURPOSE: To evaluate the safety, pharmacokinetics, and biologic effect of multiple doses of the chimeric anti-CD20 monoclonal antibody (mAb) IDEC-C2B8 in patients with relapsed B-cell lymphoma. PATIENTS AND METHODS: Twenty patients with relapsed low-grade (n = 15) or intermediate-/high-grade (n = 5) lymphoma received weekly infusions times four of 125 mg/m2 (n = 3), 250 mg/m2 (n = 7), or 375 mg/m2 (n = 10) of IDEC-C2B8. RESULTS: Infusional side effects during the initial infusion were mainly grade I/II fever, asthenia, chills, nausea, rash, and urticaria. More serious events were rare. Peripheral-blood B cells were rapidly depleted and slowly recovered over 3 to 6 months. There was no change in mean immunoglobulin (Ig) levels. Antibody serum half-life (and maximum concentration [Cmax]) generally increased between the first and fourth infusions (33.2 hours v 76.6 hours, respectively) following the 375-mg/m2 doses. Six of 18 assessable patients had a partial remission (PR), with a median time to disease progression of 6.4 months (range, 3 to 21.7). Minor responses (MRs) were observed in five patients and progressive disease (PD) in seven. Tumor responses occurred in peripheral blood, bone marrow (BM), spleen, bulky lymph nodes, and extranodal sites, and in patients who had relapsed following high-dose myeloablative chemotherapy. Six of 14 patients (40%) with a low-grade histology responded. Four of six with bulky disease had a PR. CONCLUSION: IDEC-C2B8 chimeric anti-CD20 mAb therapy is well tolerated and has clinical activity in patients with relapsed B cell lymphoma. The 375-mg/m2 dose has been selected for a phase II trial in patients with relapsed low-grade or follicular B-cell lymphoma. PMID- 9336367 TI - 1997 update of recommendations for the use of hematopoietic colony-stimulating factors: evidence-based, clinical practice guidelines. American Society of Clinical Oncology. PMID- 9336368 TI - Reed-Sternberg cells in spinal fluid. PMID- 9336369 TI - Hypofractionation, not rapid-fractionation. PMID- 9336370 TI - Delayed administration of dexrazoxane provides cardioprotection against anthracyclines in breast cancer or acute myeloid leukemia. PMID- 9336371 TI - Essential hypertension in African Caribbeans associates with a variant of the beta2-adrenoceptor. AB - Populations of West African ancestry dwelling in Western communities exhibit greater prevalence of human essential hypertension and higher rates of end-organ damage. The sympathetic nervous system influences cardiac output, vascular tone, renal sodium reabsorption, and renin release and could be implicated in enhanced vascular responsiveness observed in African hypertensives. Such an effect could arise from genetic variants that alter agonist response of alpha-adrenoceptors, leading to enhanced vasoconstriction, or attenuate beta2-adrenoceptor-mediated vasodilatation. Indeed, there is evidence of a blunted vasodilator response to the beta-agonist isoprenaline in African Americans. A variant of the beta2 adrenoceptor gene that encodes glycine rather than arginine at position 16 (Arg16 ->Gly) has been shown to confer exaggerated agonist-mediated receptor downregulation, which might attenuate vasodilator response. One hundred thirty six unrelated hypertensives and 81 unrelated normotensives of African Caribbean origin were identified from primary care on the island of St Vincent. Genomic DNA from these subjects was analyzed for the presence of the Gly16 and Arg16 alleles by using an allele-specific polymerase chain reaction method. We report strong support for association of the prodownregulatory glycine 16 variant of the beta2 adrenoceptor gene with hypertension in African Caribbeans from St Vincent and the Grenadines (chi2=18.9, P=.000014, 1 df). This observation, coupled with reports of attenuated vasodilator responses to beta-agonists among people of West African ancestry, may provide a mechanism for enhanced vascular reactivity and identify a candidate gene for hypertension in this ethnic group. PMID- 9336372 TI - Systolic function in hypertensive men with concentric remodeling. AB - Hypertensive patients with concentric remodeling (relative wall thickness > or = 0.45 and normal left ventricular [LV] mass index) may have poor outcomes. It is unclear whether systolic function abnormalities, shown to be present in some patients with concentric LV hypertrophy (increased LV mass index and relative wall thickness > or = 0.45), are also present in patients with concentric remodeling. To assess LV pump, chamber, and myocardial function in hypertensive men with concentric remodeling, clinical and echocardiographic data of 118 hypertensive men with concentric remodeling were compared with data from 104 hypertensive men with normal relative wall thickness and normal LV mass index. Chamber function was assessed by relating endocardial fractional shortening to end-systolic circumferential stress, myocardial function was assessed by relating midwall fractional shortening to circumferential stress, and pump performance was assessed by stroke volume (Teichholz method). Compared with hypertensive men with normal relative wall thickness, concentric-remodeling patients had lower stroke volume (84 +/- 20 versus 111 +/- 20 mL, P < .001). Endocardial shortening was no different between the two groups (38 +/- 7% versus 40 +/- 7%, P=NS), but midwall shortening was lower in patients with concentric remodeling (20 +/- 3% versus 22 +/- 3%, P < .001), despite lower end-systolic stress (81 +/- 25 versus 117 +/- 37 g/cm2, P < .001). Endocardial and midwall stress-shortening regression plots classified 28% and 42%, respectively, of the concentric remodeling patients below the fifth percentile of hypertensive patients with normal geometry. These data indicate that indexes of chamber and myocardial function are lower than those observed in hypertensive patients with normal geometry. Thus, indices of chamber, myocardial, and pump performance indicate potential abnormalities in systolic function in men with concentric remodeling. PMID- 9336373 TI - Cardiovascular reactivity to stress and left ventricular mass in youth. AB - We studied the relationships of cardiovascular reactivity during mental stress with left ventricular mass index in a group of prepubertal children 8 to 10 years old and in a group of peripubertal or postpubertal adolescents 15 to 17 years old. One hundred fifteen participants, varying in age group, sex, and race (black and white), took part in a laboratory stress protocol consisting of a reaction time task, a mirror tracing task, a cold forehead challenge, and a stress interview. Cardiovascular measures included blood pressure and heart rate, as well as cardiac output, stroke volume, total peripheral resistance, and preejection period obtained noninvasively with impedance cardiography. Measures of left ventricular mass were made by echocardiography. Results indicated that across all participants, left ventricular mass index was associated with cardiovascular responses during the mirror tracing and cold forehead tasks, especially with those responses reflecting increased vasoconstriction. Subgroup analyses showed that these associations were significant for males and sometimes adolescents but not for females and children. As mirror tracing and cold forehead tasks most consistently produce alpha-adrenergic activation, the results suggest a model in which vasoconstriction due to mental stress is related to increased left ventricular mass in susceptible individuals, even at a young age. PMID- 9336374 TI - Can trasmitral Doppler E-waves differentiate hypertensive hearts from normal? AB - Physiological models of transmitral flow predict E-wave contour alteration in response to variation of model parameters (stiffness, relaxation, mass) reflecting the physiology of hypertension. Accordingly, analysis of only the E wave (rather than the E-to-A ratio) should be able to differentiate between hypertensive subjects and control subjects. Conventional versus model-based image processing methods have never been compared in their ability to differentiate E waves of hypertensive subjects with respect to age-matched control subjects. Digitally acquired transmitral Doppler flow images were analyzed by an automated model-based image processing method. Model-derived indexes were compared with conventional E-wave indexes in 22 subjects: 11 with hypertension and echocardiographically verified ventricular hypertrophy and 11 age-matched nonhypertensive control subjects. Conventional E-wave indexes included peak E, E, and acceleration and deceleration times. Model-based image processing-derived indexes included acceleration and deceleration times, potential energy index, and damping and kinematic constants. Intergroup comparison yielded lower probability values for model-based compared with conventional indexes. In the subjects studied, Doppler E-wave images analyzed by this automated method (which eliminates the need for hand-digitizing contours or the manual placement of cursors) demonstrate diastolic function alteration secondary to hypertension made discernible by model-based indexes. The method uses the entire E-wave contour, quantitatively differentiates between hypertensive subjects and control subjects, and has potential for automated noninvasive diastolic function evaluation in large patient populations, such as hypertension and other transmitral flow velocity-altering pathophysiological states. PMID- 9336375 TI - Molecular mechanism of angiotensin II type I and type II receptors in cardiac hypertrophy of spontaneously hypertensive rats. AB - We administered angiotensin (Ang) II receptor type 1 (AT1) blockade (losartan; 10 or 40 mg/kg per day), type II receptor (AT2) blockades (PD123319; 100 mg/kg per day), or angiotensin-converting enzyme (ACE) inhibitor (enalapril; 30 mg/kg per day) to spontaneously hypertensive rats (SHR) from 10 to 20 weeks of age. At the end of the treatment, high doses of losartan and enalapril significantly reduced the arterial systolic blood pressure compared with the untreated SHR to the level of WKY rats. But low doses of losartan and PD123319 were without effect. High doses of losartan and enalapril also significantly reduced both the left ventricular (LV) weight and the ratio of LV to body weight compared with the untreated SHR, which were still larger than that of WKY rats. However, the collagen concentration of SHR treated with high doses of losartan or enalapril was completely reduced to the level of WKY rats. Using reverse transcription polymerase chain reaction, we examined the mRNA expression for ACE, AT1, and AT2 in experimental animals. The enhanced AT1 mRNA expression was significantly decreased in the SHR treated with a high dose of losartan or PD123319 compared with the untreated SHR. The level of ACE mRNA was also decreased in the SHR treated with a high dose of losartan or enalapril. The level of AT2 mRNA was not significantly different between the Wistar-Kyoto rats and the SHR; however, this expression was decreased significantly after the treatment with a high dose of losartan or PD123319. These results indicate that AT1 receptor and ACE, but not AT2 receptor, play a crucial role in the remodeling of matrix tissue but a smaller role in the development of the hypertrophy of LV myocyte in SHR and that the LV/body weight changes do not fully account for the complete suppression of hypertension. PMID- 9336376 TI - Broadband spectral analysis of blood pressure and heart rate variability in very elderly subjects. AB - Systolic blood pressure (SBP) variability is increased and R-R interval variability is reduced in the elderly. Little is known, however, about how SBP and R-R interval variabilities change in the very elderly. More important, however, it is not known which frequency components of SBP and R-R interval variability are affected significantly. We addressed this issue in subjects older than 70 years by broadband spectral analysis, which allows all variability components from the lowest to the highest frequency to be considered. In 20 very elderly normotensive subjects (mean +/- SD age, 78.1 +/- 6.8 years) and 28 normotensive adult subjects (36.1 +/- 7.1 years), noninvasive finger blood pressure and R-R intervals were recorded continuously for 30 minutes in the supine position and 15 minutes in the upright position. SBP and R-R interval power spectral densities were computed over the entire frequency region between 0.005 Hz (0.007 Hz in the upright position) and 0.5 Hz. Overall SBP variability (SD) was greater and overall R-R interval variability was less in very old subjects than in adult subjects. All spectral R-R interval powers were reduced significantly in very elderly individuals. The spectral SBP powers were greater in the very elderly group than in the adult group only in the very-low-frequency range (<0.04 Hz). This was true in the supine and the standing positions. With subjects in the standing position, the shape of the broadband spectra differed in the very old and adult subjects because in the former group the increase in SBP and R-R interval power around 0.1 Hz that was seen in the latter was blunted. Therefore, in very elderly subjects a reduction in overall R-R interval variability is accounted for by a reduction in all of its frequency components. The accompanying increase in overall BP variability, however, results from a nonhomogeneous behavior of its frequency components, which consists of an increase in the very low frequency and a concomitant reduction in the higher frequency powers. The mechanisms responsible for these changes may be complex, but at least they may in part reflect the baroreflex impairment and autonomic dysfunction that characterize aging. PMID- 9336377 TI - Diastolic pressure underestimates age-related hemodynamic impairment. AB - It has been hypothesized that as large arteries become more rigid with age, the pattern of hypertension changes from diastolic to systolic. Thus, diastolic blood pressure (DBP) may lose its ability to reflect the increase in vascular resistance with age. To assess this, we studied the age-related changes in blood pressure pattern and its steady-state and pulsatile determinants. We performed an epidemiological analysis based on a national survey of 10,462 subjects from Argentina. A hemodynamic analysis (impedance cardiography) was then carried out in 636 consecutive hypertensive patients (age, 25 to 74 years). Whereas the rate of increment in the prevalence of mild to moderate hypertension (MMH) reached a plateau after the sixth decade, isolated and borderline systolic forms of hypertension began a steep and sustained rise. Among patients with MMH, DBP remained stable from the third to the seventh decade, whereas SBP maintained a sustained increase. Despite similar DBP, the systemic vascular resistance index increased 47% (P<.01) and the cardiac index decreased 27% (P<.01), whereas the ratio of stroke volume to pulse pressure, an index of arterial compliance, decreased 45% (P<.01). However, there were no significant differences between older patients with MMH and those with isolated systolic hypertension in the level of SBP, vascular resistance, stroke volume, and cardiac index. Compared with age-matched normotensive control subjects, the ratio of stroke volume to pulse pressure was much more reduced in isolated systolic hypertension (48%) than in MMH (30%). In summary, the present study, carried out in a large sample of hypertensive subjects with a wide age range, showed a simultaneous impairment in vascular resistance and arterial compliance associated with aging in different patterns of hypertension. The magnitude of these changes, with opposite effects on DBP but additive effects on SBP, suggests that a hemodynamic mechanism could determine the transition in the prevalence of diastolic hypertension toward a systolic pattern of hypertension with aging. Also, the results suggest that SBP, but not DBP, is a reliable indicator of the underlying hemodynamic abnormalities (high resistance and low arterial compliance) in the elderly. PMID- 9336378 TI - Anatomic heterogeneity of vascular aging: role of nitric oxide and endothelin. AB - We investigated the effects of aging, a cardiovascular risk factor, on vascular function with regard to endothelial nitric oxide synthase (eNOS), superoxide dismutase (SOD), and endothelin (ET-1) in aorta and femoral artery of the rat. Concentration-response curves to acetylcholine, calcium ionophore A23187, norepinephrine, ET-1, big endothelin, sodium nitroprusside, and exogenous SOD were obtained. Expression of eNOS mRNA was analyzed by reverse-transcription polymerase chain reaction, SOD activity was assessed using a chemiluminescence based cytochrome c assay, and ET-1 plasma concentrations were measured by radioimmunoassay. In aorta of old rats, relaxations to acetylcholine and calcium ionophore A23187, basal NO release, and expression of eNOS mRNA in aortic endothelial cells were reduced (P<.05). In femoral arteries, relaxations to acetylcholine were preserved, whereas basal release of NO was attenuated (P<.05). Aging selectively increased contractions to norepinephrine and functional endothelin converting enzyme activity and attenuated contractions to ET-1 in aortas but not femoral arteries. Vascular SOD activity was higher in the femoral artery (P<.05) and unaffected by aging. Plasma ET-1 levels increased and plasma SOD activity decreased with age (P<.05). Aging was associated with an anatomic heterogeneity of endothelial dysfunction, functional endothelin converting enzyme activity, and vascular SOD activity. Vascular function was impaired in the aorta but not the femoral artery, which may be related to lower eNOS mRNA expression and SOD activity. These data suggest differential regulation of the vascular aging process that may contribute to the anatomic heterogeneity of atherosclerosis. PMID- 9336379 TI - Endothelin mediation of insulin and glucose-induced changes in vascular contractility. AB - Although the prevalence of hypertension in diabetic patients is high and many factors participate, hyperinsulinemia cannot be discarded as a contributing factor. Insulin could act directly on smooth muscle altering intracellular calcium levels that mediate contraction and glucose transport or could induce the secretion of endothelin by the endothelial cells lining the vessels. The aim of the present report was to study the effect of different glucose and insulin concentrations on rat vascular smooth-muscle contractile characteristics and to determine whether insulin effects are mediated by endothelin. Femoral arteries obtained from Wistar rats were placed in an in vitro chamber and superfused with different glucose and/or insulin solutions. The contractile response to KCl 80 mmol/L, measured by the force generated, showed a significant decrease with high extracellular glucose concentrations (11 mmol/L). Insulin caused a dose-dependent increase in arterial contraction induced by KCl. This increase was significant when arteries were stimulated with 80 mmol/L KCl in the presence of 5.5 mmol/L glucose, but when 40 mmol/L KCl was used, an increase was observed with both 5.5 and 11 mmol/L glucose. The insulin-induced contraction was significantly reduced in the presence of hyperimmune anti-endothelin serum and in the presence of endothelin receptor ET(A) and ET(B) antagonists PD 151,242 and BQ-788, respectively. These results suggest that hyperinsulinemia and hyperglycemia may contribute to hypertension in diabetes and that responses to insulin are mediated partially by endothelin, thus explaining why non-insulin-dependent diabetes mellitus patients show an increase in arterial pressure before the onset of nephropathy. PMID- 9336380 TI - Reversal of endothelin-1 release by stimulation of endothelial alpha2 adrenoceptor contributes to cerebral vasorelaxation. AB - Agonists acting on the vascular endothelium can modulate the release of a number of factors that interact with the surrounding smooth muscle cells and influence their tone. One such factor is the vasoconstricting agent endothelin-1 (ET-1), which has been implicated in several disease states, including stroke. However, very little is known about the physiological role of ET-1 in the cerebral circulation. We demonstrate that activation of alpha2-adrenoceptors in human pial artery endothelial cells reduces both constitutive and agonist-stimulated release of immunoreactive ET-1. That this has physiological relevance is supported by our demonstration that in segments of rabbit middle cerebral arteries, alpha2 adrenoceptor activation reduces the release of endothelium-derived ET-1 and causes an endothelium-dependent relaxation. The adrenoceptor-dependent relaxation was not blocked by combined addition of indomethacin and N omega-nitro-L-arginine in 25 mmol/L KCl-depolarizing physiological solution but was selectively antagonized by a subthreshold concentration of exogenous ET-1. Our data suggest that activation of endothelial alpha2-adrenoceptor would favor a decrease in ET-1 production and possibly promote vascular relaxation. PMID- 9336381 TI - A live-cell assay for studying extracellular and intracellular endothelin converting enzyme activity. AB - Endothelin-1 (ET-1) is formed from its precursor preproET-1 via the cleavage of the intermediate bigET-1 by endothelin-converting enzyme (ECE-1). However, the subcellular site at which this step occurs is not clear: It could occur intravesicularly along the secretory pathway or bigET-1 might be released and processed extracellularly. To address this point, we have developed an integrated autocrine system that uses a recombinant Chinese hamster ovary (CHO) luciferase reporter cell line that permanently expresses the human ET(A) receptor. Into these cells we transiently transfected human ECE-1a cDNA, either together with the human preproET-1 cDNA (as an endogenous source of bigET-1), or alone (in which case exogenous bigET-1 was added). Phosphoramidon inhibited the conversion of exogenous bigET-1 (IC50 = 5 to 30 micromol/L) much better than that of endogenous bigET-1 (IC50 > 1 mmol/L). Both conversions showed similar high yields (20% to 100%) that depended on the amount of ECE-1a expressed. Thus, ECE-1a has two equally relevant activities in this recombinant system for CHO cells: (1) an intracellular, probably intravesicular activity, corresponding to the ECE-1a mediated step of ET-1 biosynthesis and (2) an extracellular activity at the plasma membrane. If this is also the case for endothelial cells, ECE-1a inhibitors would have to cross the plasma and vesicle membranes to be effective. The present system could be useful for screening such inhibitors. PMID- 9336382 TI - Cold pressor test raises serum concentrations of ICAM-1, VCAM-1, and E-selectin in normotensive and hypertensive patients. AB - In patients with essential hypertension, elevated soluble E-selectin (sE selectin) levels may indicate endothelial cell injury or activation. We therefore sought to ascertain whether arterial blood pressure increased by the cold pressor test can modify serum concentrations of sE-selectin and other soluble forms of adhesion molecules, such as soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1), or the expression of any adhesion molecules in circulating monocytes and lymphocytes. Our findings show that levels of sE-selectin, sVCAM-1, and sICAM-1 are higher in patients with essential hypertension than in normotensive subjects (sICAM-1, 380 +/- 52 versus 262 +/- 96 ng/mL, P<.05; sVCAM-1, 720 +/- 52 versus 625 +/- 38 ng/mL, P<.05; and sE-selectin, 75 +/- 21 versus 61 +/- 22 ng/mL, P<.05). Furthermore, in normotensive and hypertensive patients, the cold pressor test caused an increase in serum concentrations of sICAM-1, sVCAM-1, and sE-selectin, but it did not cause changes in the expression of adhesion molecules in circulating monocytes and lymphocytes. High arterial blood pressure may therefore increase the production of serum adhesion molecules, probably through endothelial activation. PMID- 9336383 TI - Moderately obese, insulin-resistant women exhibit abnormal vascular reactivity to stress. AB - To define the hemodynamic implications of insulin resistance (IR), we compared 10 normotensive, insulin-resistant women who had abnormal glucose tolerance tests with 10 age-matched healthy normotensive women with normal glucose tolerance tests with respect to mental arithmetic and handgrip responses. Hemodynamic variables obtained at baseline and during stress included heart rate, blood pressure, cardiac output, and systemic vascular resistance. The IR group weighed more (84 versus 66 kg). Screening BP was similar (123/72 versus 120/68 mm Hg, P=NS) between groups although baseline diastolic BP at testing day was higher in the IR group than control group (75 versus 65 mm Hg, P<.05). The IR group showed a significantly greater increase in systolic (18% versus 10%, P<.O1) and diastolic (24% versus 12%, P<.01) blood pressure responses to mental stress than the control group. During mental stress, the control group demonstrated increased cardiac output (1.4 L/min) and decreased systemic vascular resistance (-120 dyne x s x cm[-5]), whereas IR subjects demonstrated increased systemic vascular resistance (119 dyne x s x cm(-5); group difference, P<.02) with only a small increase in cardiac output (0.5 L/min). Handgrip also caused a greater increase in systemic vascular resistance in the IR group (252 versus 64 dyne x s x cm(-5), P<.05), with a correspondingly greater increase in blood pressure than control subjects. Baseline blood pressure was correlated with weight (r=.41, P<.02) and stress blood pressure with fasting insulin (r=.51, P<.001) and glucose-to-insulin ratio (r= -.55, P<.001). We conclude that insulin resistance is associated with an exaggerated blood pressure response to stress; an enhanced vasoconstriction to stress may mediate this response. This hyperreactivity may be a marker for future hypertension in obese, normotensive, hyperinsulinemic individuals. PMID- 9336384 TI - Genetic isolation of a chromosome 1 region affecting blood pressure in the spontaneously hypertensive rat. AB - Recent linkage studies in the spontaneously hypertensive rat (SHR) suggest that a blood pressure regulatory gene or genes may be located on rat chromosome 1q. To investigate this possibility, we replaced a region of chromosome 1 in the SHR (defined by the markers D1Mit3 and Igf2) with the corresponding chromosome segment from the normotensive Brown-Norway (BN) strain. In male SHR congenic rats carrying the transferred BN chromosome segment, 24-hour average systolic and diastolic blood pressures were significantly lower than in male progenitor SHR. Polymerase chain reaction genotyping using 60 polymorphic microsatellite markers dispersed throughout the genome confirmed the congenic status of the new strain designated SHR.BN-D1Mit3/Igf2. These findings provide direct evidence that a blood pressure regulatory gene exists on the differential segment of chromosome 1 that is sufficient to decrease blood pressure in the SHR. The SHR.BN-D1Mit3/Igf2 congenic strain represents an important new model for fine mapping and characterization of genes on chromosome 1 involved in the pathogenesis of spontaneous hypertension. PMID- 9336385 TI - Tissue angiotensinogen gene expression induced by lipopolysaccharide in hypertensive rats. AB - There is now convincing evidence that various tissues express their own tissue renin-angiotensin system, which may be regulated independently of the systemic renin-angiotensin system. However, little information is available on the regulation of the tissue renin-angiotensin system. We investigated the regulation of tissue angiotensinogen gene expression with respect to the development of hypertension. We measured basal and lipopolysaccharide-stimulated plasma angiotensinogen concentrations by radioimmunoassay and examined the expression of tissue angiotensinogen by Northern blot analysis in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) at 4 and 13 weeks of age. Basal plasma angiotensinogen concentration in SHR was comparable to that in WKY at 4 weeks of age and was significantly higher than that in WKY at 13 weeks of age. Lipopolysaccharide induced a significant increase in plasma angiotensinogen concentration in both WKY and SHR at 4 and 13 weeks of age. At 4 weeks of age, the basal levels of angiotensinogen mRNA in the liver, fat, adrenal, and aorta were higher in WKY than in SHR. At 13 weeks of age, the basal levels of angiotensinogen mRNA in the fat, adrenal, aorta, spleen, and kidney were higher in WKY than in SHR, while that in the liver did not differ significantly between the two strains. At 4 weeks of age, pretreatment with lipopolysaccharide increased the angiotensinogen mRNA levels in the liver, fat, adrenal, and aorta in both WKY and SHR. At 13 weeks of age, pretreatment with lipopolysaccharide increased the angiotensinogen mRNA levels in the liver, aorta, and adrenal; decreased those in the spleen; and had no effect in the kidney in both WKY and SHR. Interestingly, lipopolysaccharide increased the angiotensinogen mRNA level in fat only in SHR, with no effect in WKY, at 13 weeks of age. Lipopolysaccharide stimulated tumor necrosis factor-a mRNA expression in fat of WKY and SHR, and the increase in tumor necrosis factor-alpha mRNA level in SHR was significantly greater than that in WKY. Therefore, the increased tumor necrosis factor-alpha mRNA expression may be involved in the increased lipopolysaccharide-induced expression of angiotensinogen gene in fat of SHR at 13 weeks of age. These data suggest that the transcriptional and probably posttranscriptional regulation of angiotensinogen mRNA differs between SHR and WKY, that the regulation of angiotensinogen gene expression is tissue-specific, and that the altered expression of the angiotensinogen gene may be involved in the development of hypertension. PMID- 9336386 TI - Enalapril and renal function in hypertensive rats transgenic for mouse renin gene. AB - We examined the effect of long-term enalapril treatment on renal function and histology in the monogenetically hypertensive TGR(mRen2)27 rat strain. Untreated transgenic rats had significantly (P<.01) higher blood pressures than treated transgenic and control animals throughout the study. Urinary nitric oxide metabolite excretion was significantly lower in young transgenic rats and rose with enalapril, suggesting abnormal TGR nitric oxide production and its correction by enalapril. Converting enzyme inhibition produced preferential preglomerular vasodilatation and increased renal blood flow (6.5 +/- 0.5 versus 9.0 +/- 0.7 mL/min per gram kidney weight, P<.05) without altering whole-kidney and single-nephron glomerular filtration rates in TGR(mRen2)27. Glomerular capillary pressure fell modestly in treated transgenic animals (54 +/- 1 versus 50 +/- 1 mm Hg, P<.05). These hemodynamic changes were associated with reductions in albuminuria (59 +/- 6 versus 9 +/- 2 mg/d, P<.01) and glomerulosclerosis in TGR. However, urinary albumin excretion (15 +/- 3 versus 3 +/- 1 mg/d, P<.05) and glomerulosclerosis also declined in treated control animals in the absence of significant alterations in glomerular hemodynamics. The mechanism of the beneficial effect of enalapril on the TGR(mRen2)27 kidney is unclear but could involve either control of hypertension or suppression of the intrarenal renin angiotensin system. PMID- 9336387 TI - Regulation of 12-lipoxygenase by cytokines in vascular smooth muscle cells. AB - Increasing evidence suggests that cytokines such as interleukin-1beta (IL-1), IL 4, and IL-8 may play an important role in the chronic inflammation and cellular growth observed in cardiovascular diseases. The lipoxygenase (LO) pathway of arachidonate metabolism has also been related to the pathology of hypertension and atherosclerosis. LO products have chemotactic, hypertrophic, and mitogenic effects in vascular cells, and the LO enzyme has been implicated in the oxidation of LDL. Furthermore, earlier studies have shown that vascular smooth muscle cell (VSMC) growth factors such as angiotensin II and platelet-derived growth factor can increase LO activity and expression in VSMCs. In the present study, we have examined whether vasoactive and inflammatory cytokines such as IL-1, IL-4, and IL 8 can modulate 12-LO activity and expression in porcine VSMCs and also whether they have growth-promoting effects in these cells. Treatment of porcine VSMCs with these cytokines led to significant increases in the levels of a cell associated 12-LO product, 12-hydroxyeicosatetraenoic acid, as well as intracellular 12-LO enzyme activity. Furthermore, each of these cytokines led to a dose-dependent increase in 12-LO mRNA expression (333-base pair PCR product) as well as 12-LO protein expression (72 kD). In addition, all three interleukins could induce significant increases in VSMC DNA synthesis as well as proliferation. These results suggest that these cytokines have mitogenic effects in VSMCs and are also potent positive regulators of the 12-LO pathway. Thus, enhanced 12-LO activity and expression may be a key mechanism for cytokine induced VSMC migration and proliferation. PMID- 9336388 TI - NHE-1 protein in vascular smooth muscle and lymphocytes from the spontaneously hypertensive rat. AB - The present study examined the abundance of NHE-1 protein in cultured vascular smooth muscle cells (VSMCs), freshly isolated thymocytes, and fresh aortic tissue from spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) rats. Two sets of affinity-purified antibodies (Ab[765-778] and Ab[698-711]) against different epitopes of the NHE-1 isoform of the Na+-H+ antiporter were used. Each set of antibodies recognized a major protein band at 105 to 110 kD that was more abundant in protein lysates prepared from cultured VSMCs from the SHR than those from WKY rats (Ab[765-778] 0.047 +/- 0.011 vs 0.010 +/- 0.002 O.D. units/10 microg protein, P<.001 for SHR and WKY, respectively; and Ab(698-711) 0.173 +/- 0.026 vs 0.087 +/- 0.028 O.D. units/10 microg protein, P<.05, for SHR and WKY, respectively). The increase in NHE-1 protein abundance in cultured VSMCs from the SHR was associated with a greater Vmax of the Na+-H+ antiporter as compared to those from WKY rats (17.93 +/- 2.07 vs 8.16 +/- 1.05 mmol H+/min, P<.001, respectively). In contrast to cultured VSMCs, there was no difference in the relative abundance of NHE-1 protein in fresh aortic tissue (0.075 +/- 0.018 vs 0.083 +/- 0.017 O.D. units/10 microg protein, from SHR and WKY, respectively) or in freshly isolated thymocytes (0.158 +/- 0.046 vs 0.226 +/- 0.054 O.D. units/10 microg protein, from SHR and WKY, respectively). We conclude that the increase in the Vmax of the Na+-H+ antiporter in cultured VSMCs from the SHR, compared to those from WKY rats, is due, at least in part, to increased levels of NHE-1 protein. PMID- 9336389 TI - Ouabain-like factor quantification in mammalian tissues and plasma: comparison of two independent assays. AB - The resolution of controversies that concern the detectability of an endogenous ouabain-like factor (OLF) in mammalian tissues and plasma was approached by the application of a standardized method for its extraction and quantification. Two independent assays were used to quantify the OLF: (1) a radioimmunoassay, which used a polyclonal anti-ouabain antiserum, and (2) a radioenzymatic assay based on the inhibition of dog kidney Na+,K+-ATPase. Plasma and tissues were obtained from the Milan hypertensive strain (MHS) and the Milan normotensive strain (MNS) of rats and from healthy human volunteers. Results indicate that (1) a single high performance liquid chromatography (HPLC) fraction identical to that of ouabain was identified by both assay methods in the rat hypothalamus and hypophysis and in both rat and human plasma; (2) dilution curves of OLF and standard ouabain were parallel and with a similar Kd, both in radioimmunoassay (3 nmol/L) and ATPase assay (14 nmol/L); (3) after HPLC, OLF was similarly quantified by the two methods in the hypothalamus, hypophysis, adrenals, and plasma of rats and in human plasma; (4) OLF was present in larger amounts in the hypothalamus, hypophysis, and plasma of MHS rats than that of MNS rats; (5) the HPLC fraction of human plasma was quantified similarly by both assays (range, 60 to 150 pmol/L); (6) recovery of standard ouabain in pre-HPLC plasma extracts was approximately 90%; and (7) pre-HPLC OLF concentrations in human plasma ranged between 0.05 and 0.75 nmol/L. Rat cerebral tissues and both rat and human plasma contained measurable amounts of OLF, which were quantified similarly by radioimmunoassay and ATPase assay, both before and after HPLC fractionation. The increased MHS tissue and plasma levels of OLF are in keeping with the pathogenetic role of this factor in MHS hypertension. PMID- 9336390 TI - A peptide released by pepsin from kininogen domain 1 is a potent blocker of ANP mediated diuresis-natriuresis in the rat. AB - A 20-amino acid peptide, KYEIKEGDCPVQSGKTWQDC (PU-D1), released by pepsin hydrolysis of LMW kininogen domain 1 was tested for its ability to antagonize the diuretic and natriuretic effect of ANP(103-125) in anesthetized rats. A single dose of 10.8 or 21.6 pmol (25 or 50 ng) PU-D1 given intravenously or into the duodenal lumen suppressed the diuresis-natriuresis induced by 209 pmol (500 ng) ANP by 43% to 59% and 69% to 96%, respectively. None of the doses tested (2.16 to 432 pmol, 5 ng to 1 microg) modified systemic blood pressure. Strikingly, a single IV dose of 10.8 pmol PU-D1 blocked the action of ANP for more than 3 hours. ANP blockade by PU-D1 was annulled completely by the bradykinin (BK) B2 receptor inhibitor Hoe 140. On a molar basis, PU-D1 is more effective than BK and kinins of 15, 16, and 18 amino acids for blocking the ANP-mediated diuresis natriuresis. As with BK and other kinins, the inhibitory effect of Pu-D1 on ANP is obtained only within a small range of picomol doses. A single dose of 2.16 or 4.32 pmol PU-D1 or 47 pmol (50 ng) BK is ineffective against ANP if injected alone. However, when both substances are administered concomitantly at these subthreshold doses, they totally suppress ANP-induced diuresis-natriuresis. These results raise the question of whether PU-D1, released from kininogen domain 1, either alone or associated with BK, may interact with ANP in the regulation of urinary water and electrolyte excretion in physiological and pathological conditions. PMID- 9336391 TI - Structure and function of small arteries in salt-induced hypertension: effects of chronic endothelin-subtype-A-receptor blockade. AB - The involvement of endothelin in salt-induced hypertension is unclear. In the Dahl rat model, we studied the effects of a selective endothelin-subtype A (ET[A]) receptor antagonist, LU135252, on blood pressure, vascular structure, and function. Dahl salt-sensitive and salt-resistant rats were treated for 8 weeks with 4% NaCl alone or in combination with LU135252 taken orally (60 mg/kg per day). The geometry and reactivity of basilar and mesenteric arteries were studied in vitro under perfused and pressurized conditions using a video dimension analyzer. Chronic salt administration increased systolic blood pressure by 37 +/- 3 mm Hg and media-lumen ratio of the basilar and mesenteric arteries in salt sensitive rats (P<.05). These structural changes were caused by eutrophic remodeling in basilar and hypertrophic remodeling in mesenteric arteries. Endothelium-dependent relaxations to acetylcholine and contractions to endothelin 1 were impaired in mesenteric arteries of salt-sensitive rats on a high NaCl diet. LU135252 prevented part of the increase in systolic blood pressure and structural and functional alterations but increased plasma endothelin 1 levels (P<.05 versus salt-treated, saltsensitive rats). LU135252 had no effect on these parameters in salt-resistant rats. These findings suggest that the long-term pressor effect of salt administration is mediated in part by the action of endogenous endothelin acting via ET(A) receptors. Thus, chronic ET(A) receptor blockade may be useful therapeutically to lower arterial pressure and prevent endothelial dysfunction and hypertrophic remodeling of resistance arteries in salt-sensitive forms of hypertension. PMID- 9336392 TI - Quinaprilat induces arterial vasodilation mediated by nitric oxide in humans. AB - The beneficial therapeutic effects of angiotensin-converting enzyme (ACE) inhibitors are the result of reduced angiotensin II formation and possibly also of an accumulation of bradykinin that is inactivated by ACE. In particular, recently developed ACE inhibitors with tissue-penetrating properties, such as quinaprilat, may exert vascular effects via the bradykinin B2-receptor. To test direct arterial effects of quinaprilat and enalaprilat and to study their effects on vasodilation induced by bradykinin, venous occlusion plethysmography was used during local intra-arterial drug administration into the brachial artery in healthy volunteers. The response to bradykinin was augmented by both ACE inhibitors, but the effect of quinaprilat (3.9 nmol/min) was exclusively attributable to its direct vasodilator action. Enalaprilat (13 nmol/min) did not change baseline blood flow in the human forearm circulation. In contrast, quinaprilat significantly increased arterial flow from 3.5 +/- 0.5 to 4.6 +/- 0.7 mL/100 mL tissue/min, which was inhibited by N(G)-monomethyl-L-arginine (8 micromol/min IA). Moreover, the effect of sodium nitroprusside (0.023 to 22.9 nmol/min) was substantially attenuated during concomitant administration of quinaprilat. These results suggest that quinaprilat induces vascular effects beyond the inhibition of angiotensin II formation; it causes vasodilation by increasing vascular nitric oxide production and thereby attenuates the relaxing effect of the nitric oxide donor sodium nitroprusside. PMID- 9336393 TI - Decreased vasodilator response to isoproterenol during nitric oxide inhibition in humans. AB - The vasodilator effect of beta-adrenergic agonists has traditionally been ascribed solely to a direct effect on vascular smooth muscle. Experimental studies, however, have suggested a role of endothelium-derived nitric oxide (NO) in beta-adrenergic-mediated vasodilation. The purpose of this investigation was to determine whether NO contributes to the vasodilator effect of beta-adrenergic stimulation in humans. We analyzed the forearm blood flow response to increasing doses of isoproterenol (50, 100, and 200 ng/min), a beta-adrenoceptor agonist, during the concomitant infusion of saline or N(G)-monomethylL-arginine (L-NMMA; 4 micromol/min), a blocker of NO synthesis, in 23 normal subjects (9 men and 14 women, aged 48 +/- 7 years). The effect of L-NMMA was also assessed during infusion of sodium nitroprusside (0.8, 1.6, and 3.2 microg/min), an exogenous NO donor. Drugs were infused into the brachial artery, and forearm blood flow was measured by plethysmography. The vasodilator effect of isoproterenol was significantly blunted during the administration of L-NMMA compared with saline (maximum flow, 7.7 +/- 4 versus 11.2 +/- 5 mL x min(-1) x dL(-1), respectively; P<.001). In contrast, the vasodilator response to sodium nitroprusside was not significantly affected by the infusion of L-NMMA (maximum flow, 8.8 +/- 3.7 mL x min(-1) x dL(-1) during L-NMMA versus 8.9 +/- 3.2 mL x min(-1) x dL(-1) during saline; P=.25). These findings indicate that NO inhibition blunts the vasodilator effect of beta-adrenergic agonists in the human forearm and suggest that an abnormal response to adrenergic stimulation may occur in conditions associated with impaired NO activity. PMID- 9336394 TI - Parathyroid hormone-related protein upregulates interleukin-1beta-induced nitric oxide synthesis. AB - The effect of parathyroid hormone-related protein on interleukin-1beta-induced nitric oxide production was studied in rat vascular smooth muscle cells. Interleukin-1beta time- and dose-dependently enhanced the production of nitrite, a stable metabolite of nitric oxide. Parathyroid hormone-related protein(1-34) alone up to 10(-7) mol/L had no obvious effect, but significantly increased the cytokine-induced nitrite production. RNA analysis revealed that the synergistic effect of parathyroid hormone-related protein(1-34) resulted from a potentiation of the expression of inducible nitric oxide synthase and GTP-cyclohydrolase I, the rate-limiting enzyme in the synthesis of tetrahydrobiopterin, which is a cofactor of nitric oxide synthase. The increased nitric oxide release induced by interleukin-1beta or interleukin-1beta with parathyroid hormone-related protein(1 34) was completely inhibited by coincubation with 3x10(-3) mol/L N(G)-monomethyl L-arginine, a competitive inhibitor of nitric oxide synthase, or with 10(-3) mol/L 2,4-diamino-6-hydroxypyrimidine, an inhibitor of GTP-cyclohydrolase I. Endothelin-1 potentiated interleukin-1beta induction of nitric oxide, which might be mediated by endogenous parathyroid hormone-related protein. Neutralization of exogenous or endogenous parathyroid hormone-related protein with antibody attenuated the synergistic effect of parathyroid hormone-related protein, but did not affect interleukin-1beta induction of nitric oxide. These results suggest that locally produced parathyroid hormone-related protein acts as a synergistic regulator upregulating interleukin-1beta-induced nitric oxide synthesis in the cardiovascular system, and thereby may affect vascular tone and/or vascular remodeling after vascular injury in some pathological processes such as atherosclerosis and hypertension. PMID- 9336395 TI - Insulin acutely inhibits cultured vascular smooth muscle cell contraction by a nitric oxide synthase-dependent pathway. AB - Insulin acutely decreases contractile agonist-induced Ca2+ influx and contraction in endothelium-free cultured vascular smooth muscle (VSM) cells, but the mechanism is not known. Since it has been reported that insulin-induced vasodilation in humans is linked to nitric oxide synthase activity, we wished to determine whether insulin inhibits Ca2+ influx and contraction of cultured vascular smooth muscle cells by a nitric oxide synthase-dependent pathway. Primary cultures of endothelial cell-free VSM cells from canine femoral artery were preincubated with and without 1 nmol/L insulin for 30 minutes, and the 5 minute production of cGMP was measured. Insulin alone did not affect cGMP production, but in the presence of 10(-5) mol/L serotonin insulin stimulated cGMP production by 60%. N(G)-monomethyl-L-arginine (0.1 mmol/L), an inhibitor of nitric oxide synthase, inhibited the conversion of arginine to citrulline by these cells, blocked insulin-stimulated cGMP production, and blocked the inhibition by insulin of 5-hydroxytryptamine (5-HT)-stimulated Mn+2 (a Ca2+ surrogate) influx and contraction. Insulin did not affect contraction of VSM cells grown under conditions designed to deplete the cells of tetrahydrobiopterin, an essential cofactor of nitric oxide synthase. These studies demonstrate that insulin acutely inhibits 5-HT-stimulated Ca2+ influx and contraction of endothelium-free cultured VSM cells by a nitric oxide synthase dependent mechanism. PMID- 9336396 TI - Endothelial dysfunction coincides with an enhanced nitric oxide synthase expression and superoxide anion production. AB - We investigated the effects of aortic banding-induced hypertension on the endothelium-dependent vasodilator responses in the aorta and coronary circulation of Sprague-Dawley rats. We studied the influence of hypertension on the endothelial nitric oxide synthase (NOS III) expression, assessed by Western blot and reverse transcription-polymerase chain reactions experiments, and on the superoxide anion (O2-) production. Two weeks after aortic banding, the endothelium-dependent relaxations were not altered. At this time, the expression of NOS III in the aorta and in confluent coronary microvascular endothelial cells (RCMECs) exhibited no marked changes, whereas O2- production was enhanced 1.9 fold in aortas from aortic-banded rats. Six weeks after aortic banding, the endothelium-dependent dilations were markedly impaired in the heart (50% decrease) and aorta (35% decrease). Analysis of NOS III protein and mRNA levels revealed marked increases in both aortas and confluent RCMECs (2.6- to 4-fold) from aortic-banded compared with sham-operated rats. There was no further increase in O2production in both the aorta and confluent RCMECs from aortic banded rats. An enhanced nitrotyrosine protein level was also detected in the aorta from 6-week aortic-banded rats. These findings indicate that in hypertension induced by aortic banding, an enhanced O2- production alone is not sufficient to produce endothelial dysfunction. Endothelial vasodilator hyporesponsiveness was observed only when NOS III expression and O2- production were increased and was associated with the appearance of enhanced nitrotyrosine residues. This would suggest that the development of endothelial dysfunction is linked to an overproduction of not one, but two, endothelium-derived radicals that might lead to the formation of peroxynitrite. PMID- 9336397 TI - Impaired nitric oxide- and prostaglandin-mediated responses to flow in resistance arteries of hypertensive rats. AB - In human and experimental hypertension, flow (shear stress)-induced dilation in large arteries is attenuated and resistant to nitric oxide blockade. We tested the hypothesis that a defect in nitric oxide-and/or prostaglandin-dependent flow induced dilation might occur in mesenteric resistance arteries from spontaneously hypertensive rats (SHR). We measured resistance mesenteric artery diameter in situ by intravital microscopy and simultaneously measured mesenteric arterial pressure in a collateral artery. The flow-diameter-pressure relationship was established in normotensive Wistar-Kyoto rats (WKY) and in SHR under control conditions and after endothelium removal, inhibition of nitric oxide synthesis with N omega-nitro-L-arginine methyl ester (10 micromol/L), or inhibition of prostaglandin synthesis with indomethacin (10 micromol/L). Production of prostaglandins was determined in the perfusate. Endothelium removal decreased artery diameter by 14 +/- 1.6% in WKY and 5 +/- 0.5% (P<.01 versus WKY) in SHR at a flow rate of 400 microL/min. In WKY, N omega-nitro-L-arginine methyl ester and indomethacin decreased resistance artery diameter by 12 +/- 3% (P<.001) and 5 +/- 2% (P<.01), respectively, at a flow rate of 400 microL/min; neither substance had any significant effect in SHR. In both strains, flow induced the production of 6 keto-prostaglandin F1alpha, the metabolite of prostacyclin; prostaglandin F2alpha; and thromboxane B2, the stable metabolite of thromboxane A2. Production of 6-keto-prostaglandin F1alpha and prostaglandin F2alpha was significantly lower in SHR than WKY, and TxB2 production was significantly higher in SHR than WKY. The present findings suggest that in SHR mesenteric resistance arteries, dilation in response to increases in flow was resistant to nitric oxide and prostaglandin synthesis blockade. A modification of the ratio of vasodilator to vasoconstrictor prostaglandins might be at least partly responsible for the decreased dilator response to flow in SHR. PMID- 9336398 TI - Immunohistochemically detected protein nitration indicates sites of renal nitric oxide release in Goldblatt hypertension. AB - In the kidney, nitric oxide (NO) from the macula densa (MD) is considered an integral modulator of the tubulovascular message system, whereas endothelium derived NO is a major vasorelaxing factor. The goal of the present study was to determine extracellular pathways of NO in rats with renovascular two-kidney, one clip Goldblatt hypertension (2K1C). To localize NO in the tissue, immunohistochemical detection of NO-dependent tyrosine nitration was performed using a monoclonal antibody against nitrotyrosine. Nitration of phenolic compounds such as tyrosine results from the reaction with peroxynitrite (ONOO ) formed by NO and molecular oxygen or superoxide and may therefore be used as a footprint for local release of NO. Significant nitrotyrosine immunoreactivity was detected in the extraglomerular mesangium (EGM) of the stenotic kidney in 2K1C rats, whereas in the nonclipped contralateral kidney and in control animals no signal was detected at this site. Positive staining of the EGM was paralleled by enhanced NADPH diaphorase (NADPH-d) staining of the adjacent MD, signifying increased type I nitric oxide synthase (NOS) activity in the stenotic kidney. In contrast, in the cortical vasculature selectively enhanced nitrotyrosine immunoreactivity was detected in the arteriolar wall of the nonclipped contralateral kidney, and endothelial NADPH-d signal, indicating NOS Type III activity, was enhanced in parallel. Our results suggest that in MD, stimulation of NOS in the stenotic Goldblatt kidney induces the release of NO into the EGM. From there an NO-dependent intermediate stimulus may reach the glomerular vasculature. Footprints of NO-dependent effects in the vascular smooth muscle layer of the non-clipped contralateral kidney indicate a marked vasodilatory response that may have been caused by enhanced shear stress and/or angiotensin II levels. PMID- 9336399 TI - Brain nitric oxide synthase messenger RNA in central mineralocorticoid hypertension. AB - The mechanism underlying the central hypertensinogenic effects of mineralocorticoids remains unclear. Given that nitric oxide (NO) is thought to act at autonomic sites in the brain to regulate arterial blood pressure, the effects of the potent mineralocorticoids aldosterone and 19-noraldosterone on the abundance of neuronal NO synthase (nNOS) mRNA in the brain were investigated. Wistar-Kyoto rats received a continuous intracerebroventricular infusion of aldosterone or 19-noraldosterone (5 ng/h) from an implanted osmotic minipump for 4 weeks. Total RNA was purified from microdissected tissue blocks containing the hypothalamus, dorsal medulla, rostral ventrolateral medulla, or caudal ventrolateral medulla, and changes in the abundance of nNOS mRNA were determined with a semiquantitative competitive polymerase chain reaction method. Blood pressure was significantly increased in rats 2, 3, and 4 weeks after the onset of intracerebroventricular aldosterone or 19-noraldosterone infusion compared with that in animals receiving vehicle. Subcutaneous infusion of either mineralocorticoid had no effect on blood pressure. Compared with controls, rats treated with aldosterone or 19-noraldosterone for 4 weeks showed significant decreases in the amount of nNOS mRNA in the hypothalamus and rostral and caudal ventrolateral medulla. These data suggest that reduced nNOS activity may contribute to the increase in blood pressure in rats with central mineralocorticoid-induced hypertension. PMID- 9336400 TI - Attenuation of neurogenic vasoconstriction by nitric oxide in hindlimb microvascular beds of the rat in vivo. AB - There is evidence that sympathetic nerve activity leads to endothelium-derived nitric oxide release, which in turn attenuates neurogenic vasoconstriction. Here we tested in vivo (1) whether the magnitude of the vasoconstriction induced by N(G)-nitro-L-arginine methyl ester given systemically is altered when ongoing sympathetic activity is abolished by sectioning the lumbar sympathetic trunk, and (2) whether hindlimb sympathetic vasoconstriction elicited by electrical stimulation of the lumbar sympathetic trunk is enhanced after inhibition of nitric oxide synthesis. Blood flow in the microvascular beds of hairless skin and skeletal muscle of the rat hindlimb was measured with laser Doppler flowmetry. Sectioning the lumbar sympathetic trunk resulted in an increase of blood flow in both tissues, indicating that tonic neurogenic vasoconstriction was abolished. Inhibition of nitric oxide synthesis resulted in vasoconstriction in both vascular beds. This vasoconstriction was more pronounced after abolition of sympathetic activity than with intact sympathetic supply in skin but was smaller in skeletal muscle. The vasoconstriction elicited by graded electrical stimulation of the centrally sectioned lumbar sympathetic trunk with frequencies less than 5 Hz was significantly enhanced after blockade of nitric oxide in skeletal muscle but not in skin microvasculature. These findings suggest that under physiological conditions, sympathetic nerve impulses directly promote the release of nitric oxide in skeletal muscle but not in cutaneous blood vessels. Therefore, basal nitric oxide release is probably in part dependent on sympathetic activity in skeletal muscle, whereas it appears to be mainly due to flow-dependent shear stress in hairless skin microvasculature. PMID- 9336401 TI - Role of endogenous carbon monoxide in central regulation of arterial pressure. AB - We investigated the contribution of neural mechanisms to the arterial pressure increase produced by zinc deuteroporphyrin 2,4-bis glycol (ZnDPBG), an inhibitor of endogenous carbon monoxide synthesis. The arterial baroreceptor reflex control of heart rate was examined in rats with and without ZnDPBG pretreatment (45 micromol/kg IP) by analysis of the arterial pressure-heart rate relationship during infusions of phenylephrine or sodium nitroprusside to vary arterial pressure. ZnDPBG increased arterial pressure from 110 +/- 3 to 126 +/- 2 mm Hg without eliciting bradycardia. The maximum gain of the heart rate response to changes in arterial pressure was attenuated by ZnDPBG treatment (-1.9 +/- 0.3 versus -4.8 +/- 1.0 bpm/mm Hg). The possibility that ZnDPBG elevates arterial pressure by attenuating baroreceptor reflex function was addressed by comparing the pressor response to ZnDPBG (45 micromol/kg IP) in rats with and without sinoaortic denervation. The pressor effect of ZnDPBG was similar in rats with and without arterial baroreceptor deafferentation, implying that the increase in pressure is not simply the consequence of attenuated baroreceptor reflex function per se. The possibility that ZnDPBG increases arterial pressure via an effect on the nucleus tractus solitarii (NTS) also was investigated. ZnDPBG (1 nmol in 100 nL) injected into the NTS of rats increased arterial pressure from 111 +/- 4 to 126 +/- 5 mm Hg, and this effect was reversed by an ipsilateral microinjection of carbon monoxide into the NTS. Accordingly, the pressor effect of ZnDPBG may rely on inhibition of carbon monoxide production in the NTS. This implies that carbon monoxide formed by brain heme oxygenase plays a role in the central regulation of arterial pressure. PMID- 9336403 TI - Dynamic autoregulation and renal injury in Dahl rats. AB - The Dahl salt-sensitive (Dahl S) rat develops hypertension and renal injuries when challenged with a high salt diet and has been considered to be a model of chronic renal failure. Renal injuries appear very early in life compared with the spontaneously hypertensive rat (SHR). During the course of hypertension, a gradual impairment of autoregulatory control of renal blood flow might expose the glomerular circulation to periods of elevated pressure, resulting in renal injuries in Dahl S rats. Dynamic autoregulatory capacity was assessed in Dahl S and Dahl salt-resistant (Dahl R) rats, SHR, and Sprague-Dawley rats by inducing broad-band fluctuations in the arterial blood pressure and simultaneously measuring renal blood flow. Dynamic autoregulation was estimated by the transfer function using blood pressure as the input and renal blood flow as the output. Renal morphological injuries were evaluated in Dahl S rats and SHR and were scored semiquantitatively. Dynamic autoregulation was efficient and comparable in the low-frequency range (<0.015 Hz) in Dahl R rats, SHR, and Sprague-Dawley rats. The response in Dahl S rats depended strongly on the initiation time of the high salt diet. Autoregulation was preserved during a low salt diet and in rats exposed to a late-onset hypertension of short duration, only partly preserved if the late-onset hypertension was of a longer duration, and abolished in early onset hypertension. All Dahl S rats on a high salt diet showed severe morphological changes in the kidney. In conclusion, autoregulatory capacity in the kidney of Dahl S rats is gradually impaired when rats are rendered hypertensive with a high salt diet. Renal morphological injuries develop before loss of dynamic autoregulation. Impaired autoregulation appears to be the result, not the cause, of the process that ultimately leads to renal failure in the Dahl S rat. PMID- 9336402 TI - Lovastatin prevents development of hypertension in spontaneously hypertensive rats. AB - The present study evaluated the effects of lovastatin on renal function and the development of hypertension in spontaneously hypertensive rats (SHR). Four-week old SHR were given lovastatin (10 mg/kg) or vehicle twice daily by gavage. After 4 weeks of treatment, mean arterial pressure was significantly lower in lovastatin-treated SHR (131 +/- 4 mm Hg, n=5) than in control animals (160 +/- 4 mm Hg, n=12) (P<.05). The fall in arterial pressure in lovastatin-treated rats was accompanied by changes in renal function. The slope of the relationship between arterial pressure and sodium excretion was threefold greater in lovastatin-treated SHR (n=6) than in control rats (n=6), and this was associated with significant elevations in renal medullary blood flow and renal interstitial hydrostatic pressure. Glomerular filtration rate was 17% higher in lovastatin treated SHR (n=6) than in control rats (n=6) (0.94 +/- 0.05 versus 0.81 +/- 0.07 mL/min per g of kidney weight, P<.05). The wall-to-lumen area ratio of renal arterioles was significantly reduced in lovastatin-treated SHR compared with vehicle-treated rats (0.86 +/- 0.05 versus 1.08 +/- 0.04 for vessels with inner diameters <50 microm and 0.62 +/- 0.02 versus 0.75 +/- 0.04 for vessels with inner diameters of 50 to 100 microm, P<.05). These results indicate that chronic treatment with lovastatin shifts the relations between renal medullary blood flow, renal interstitial pressure, sodium excretion, and renal perfusion pressure to lower levels of arterial pressure and attenuates the development of hypertension and renal vascular hypertrophy in SHR. PMID- 9336404 TI - CD40-CD40 ligand interaction in autoimmune disease. PMID- 9336405 TI - Ankylosing spondylitis: the dissection of a complex genetic disease. PMID- 9336406 TI - Fas/Fas ligand expression and induction of apoptosis in chondrocytes. AB - OBJECTIVE: To examine the expression of Fas/Fas ligand and the role of this ligand/receptor interaction in the regulation of apoptosis in normal human articular chondrocytes and in osteoarthritis (OA) cartilage. METHODS: Normal and OA human knee cartilage and cells isolated from these tissues were tested for Fas expression by flow cytometry. Induction of apoptosis by antibody to Fas was analyzed by DAPI staining and electron microscopy. RESULTS: Treatment of freshly isolated normal human articular chondrocytes with an agonistic Fas antibody induced apoptosis in a subpopulation (approximately 20%) of the cells. Apoptosis induced by anti-Fas was not dependent on nitric oxide (NO), and anti-Fas also did not induce NO production. Analysis of isolated cells demonstrated similar levels of Fas expression on normal and OA chondrocytes (28% and 32%, respectively). In normal articular cartilage, Fas-positive cells were located mainly in the superficial and midzones. In contrast, in fibrillated OA cartilage, surface layers were partially absent and Fas-expressing cells were also detected in the deeper layers. Fas ligand messenger RNA was not detectable in resting or activated normal or OA chondrocytes. Analysis by electron microscopy showed the nuclear and cytoplasmic changes typical of apoptosis in cultures treated with antibody to Fas. CONCLUSION: A subpopulation of chondrocytes expresses Fas and is susceptible to Fas-induced apoptosis. Fas-mediated chondrocyte apoptosis may contribute to cartilage degradation in arthritis. PMID- 9336407 TI - Periosteal new bone formation in a canine neuropathic model of osteoarthritis. AB - OBJECTIVE: To characterize, for the first time, periosteal new bone formation in a well-established canine model of accelerated osteoarthritis (OA) with features of neuropathic arthropathy. METHODS: Seven dogs underwent left L4-S1 dorsal root ganglionectomy (DRG), followed 3 weeks later by transection of the anterior cruciate ligament of the ipsilateral knee (ACLT). Eight weeks thereafter, a postmortem examination was performed to assess the severity of cartilage changes of OA and the formation of new bone on the distal femur and proximal tibia in the cruciate-deficient limb. RESULTS: As described previously, extensive full thickness ulceration of the articular cartilage was present in the unstable knee of every dog. The femoral shaft immediately proximal to the condyles in the unstable limb was consistently wider (mean +/- SD diameter 22.4 +/- 2.2 mm) than that in the contralateral limb (19.9 +/- 1.3 mm; P = 0.01). Xeroradiography and histologic examination of the distal femur revealed extensive formation of woven bone on the periosteal surfaces of the medial, lateral, and anterior aspects of the femoral shaft in the OA limb of every dog. These bony changes were not seen in radiographs of dogs that underwent DRG with the cruciate ligament left intact (n = 8) or of neurologically intact dogs that underwent ACLT (n = 7) and were examined 24 weeks after surgery. CONCLUSION: Formation of new periosteal bone on the distal femur and tibia is a feature of this model of accelerated OA that is not seen in the conventional ACLT model of OA in the neurologically intact dog. This observation suggests that interruption of sensory input from the limb may affect the regulation of osteogenesis in the mechanically unstable joint. PMID- 9336409 TI - Histologic/histochemical grading system for osteoarthritic articular cartilage: reproducibility and validity. AB - OBJECTIVE: To evaluate the reproducibility and validity of the histologic/histochemical grading system (HHGS) for osteoarthritis (OA) developed by Mankin and coworkers. METHODS: Sections of human articular cartilage from macroscopically normal and osteoarthritic femoral heads were graded by the HHGS (0-14 scale) twice by 3 observers, and intra- and interobserver reproducibility was determined. RESULTS: The exact intra- and interobserver reproducibilities ranged from 22% to 33% and from 13% to 25%, respectively. Intra- and interobserver variation ranged from -7 to 5. Although scores were generally higher in cartilage from osteoarthritic joints as compared with cartilage from normal joints, the difference reached statistical significance for only 2 observers. Additionally, the average scores from all observers were within the definitive range for OA (>5) in 30% of sections from normal joints, and in only 47% of sections from OA joints. CONCLUSION: The results indicate that the reproducibility and the validity of the HHGS are inadequate, and an improved histopathologic grading system for OA is needed. PMID- 9336408 TI - Investigation of the association of the CRTM and CRTL1 genes with radiographically evident osteoarthritis in subjects from the Rotterdam study. AB - OBJECTIVE: To investigate whether radiographically evident osteoarthritis (ROA) in 55-65-year-old men and women is associated with specific alleles or genotypes of the cartilage matrix protein (CRTM) and cartilage link protein (CRTL1) genes. METHODS: Cases were selected from a population-based study on the presence of ROA of the knee or hip. Further radiographic analysis included scoring for spine and hand ROA. Controls, selected from the same population, were free of ROA in all joints. RESULTS: The CRTM locus was significantly associated with hip ROA in men (odds ratio 0.50, 95% confidence interval 0.26-0.95). A significant association between ROA and the CRTL1 gene was not observed. CONCLUSION: These results suggest that the CRTM locus may play a role in the sex- and joint site-specific pattern of ROA development. PMID- 9336410 TI - Posttranscriptional regulation of collagenase-1 gene expression in synoviocytes by adenosine receptor stimulation. AB - OBJECTIVE: To characterize the transcriptional and posttranscriptional regulation of collagenase-1 by adenosine receptor stimulation in interleukin-1 (IL-1) stimulated fibroblast-like synoviocytes (FLS). METHODS: FLS were stimulated with IL-1 and either the nonselective adenosine agonist 5'-N-ethylcarboxamidoadenosine (NECA) or the adenylate cyclase activator forskolin. Electrophoretic mobility shift assays were performed to determine AP-1 and cAMP-responsive element binding protein (CREB) activation. Transcriptional activation was determined by transfecting HS68 dermal fibroblasts with a collagenase-chloramphenicol acetyltransferase construct. Finally, collagenase messenger RNA (mRNA) half-life was determined by activating cells in the presence of IL-1, IL-1 + NECA, or IL-1 + forskolin and culturing cells in the presence of actinomycin D. RESULTS: NECA and forskolin had no effect on AP-1 activation, c-jun or c-fos gene expression, or CREB phosphorylation. IL-1 markedly increased collagenase promoter activity, and neither NECA nor forskolin blocked this action. Studies of mRNA half-life showed that both NECA and forskolin decreased the half-life of collagenase mRNA in IL-1-stimulated FLS and HS68 cells. CONCLUSION: The findings of this study demonstrate that NECA and forskolin decrease collagenase gene expression in FLS and dermal fibroblasts due to enhanced mRNA degradation. PMID- 9336411 TI - Significant correlation between thrombospondin 1 and serine proteinase expression in rheumatoid synovium. AB - OBJECTIVE: Thrombospondin 1 (TSP1) is a potent active site inhibitor of leukocyte elastase and cathepsin G. This effect is markedly dependent on the disulfide-bond conformation of TSP1, with one isoform, TSP1(0.1), being the most potent. The aims of this study were to examine the expression of different disulfide-bonded isoforms of TSP1 in inflammatory environments in which elastase and cathepsin G are present in variable amounts, and to determine the relationship between these proteinases and their potential inhibitor. METHODS: Immunohistochemical staining and histomorphometric analysis were used to examine adjacent sections of synovial tissue from patients with rheumatoid arthritis (RA), osteoarthritis (OA), and meniscal trauma (MT), for expression of TSP1 and the TSP1(0.1) isoform, elastase, cathepsin G, and chymase. RESULTS: TSP1 localized to vessels and cells within the synovium. TSP1 expression was highly up-regulated in RA (mean density 98 cells and vessels/mm2, compared with 13/mm2 in OA and 17/mm2 in MT). The TSP1(0.1) isoform was found virtually exclusively in RA, with 44% of vascular TSP1 staining being due to the TSP1(0.1) isoform in RA, as compared with 7% in OA (P = 0.0047). Elastase- and cathepsin G-positive cells were abundant in RA, with mean densities of 106 cells/mm2 and 103 cells/mm2, respectively, compared with 2 cells/mm2 and 11 cells/mm2 in OA. There was a wide range of both TSP1 and proteinase expression within the RA group, but samples containing large numbers of elastase- and cathepsin G-positive cells also showed high expression of TSP1, especially TSP1(0.1). A strong correlation was found between elastase or cathepsin G densities and TSP1(0.1) expression in blood vessels (r = 0.86 and r = 0.76 respectively, P < 0.01). CONCLUSION: TSP1(0.1), with the most potent inhibitory activity in vitro, is specifically up-regulated in RA, and this up-regulation is in proportion to the numbers of surrounding leukocytes containing elastase and cathepsin G. One role of TSP1 may be to act as a matrix-based regulator of leukocyte-derived serine proteinases in vivo. PMID- 9336412 TI - Crucial role of interleukin-10/interleukin-12 balance in the regulation of the type 2 T helper cytokine response in reactive arthritis. AB - OBJECTIVE: To investigate whether a predominant type 1 T helper (Th1) or Th2 cytokine pattern is present in the joints of patients with reactive arthritis (ReA), and whether the cytokine pattern can be modulated by cytokines or anticytokines. METHODS: Eleven patients with ReA following infection with either Chlamydia trachomatis, Yersinia enterocolitica, or Salmonella enteritidis were investigated for the presence of Th1/Th2 cytokines in the joints. Release of the bacteria-specific cytokines interferon-gamma (IFN gamma), tumor necrosis factor alpha (TNF alpha), interleukin-10 (IL-10), and IL-4 was measured in synovial fluid mononuclear cells (SFMC) using enzyme-linked immunosorbent assay and polymerase chain reaction. In the synovial membrane, secretion of IFN gamma and IL-4 was determined by immunohistologic analysis. Cytokine regulation was studied by adding cytokines and anticytokines to the cultures. RESULTS: Upon stimulation with specific bacteria, SFMC secreted low amounts of IFN gamma and TNF alpha, but high amounts of IL-10. IL-10 was responsible for the suppression of IFN gamma and TNF alpha, as judged by the effect of adding either anti-IL-10 antibodies or exogenous IL-10 to these cultures. The addition of neutralizing anti-IL-12 to the cultures completely abolished the effects of anti-IL-10, suggesting that inhibition of the Th1-like cytokines by IL-10 is mediated through suppression of IL-12 synthesis. Exogenous IL-12 clearly enhanced IFN gamma and TNF alpha secretion. In the synovial membrane, a higher number of cells were positive for the Th2 cytokine IL-4, compared with the amount of IFN gamma-secreting cells. CONCLUSION: These data indicate that a Th2 cytokine pattern predominates in the joints of patients with ReA. Since Th1 cytokines are necessary for the elimination of ReA-associated bacteria, Th2 cytokines might contribute to bacterial persistence in the joint. Therefore, the IL-10/IL-12 balance appears to be crucial for regulation of the cytokine pattern in the joints of patients with ReA. PMID- 9336413 TI - A linkage study across the T cell receptor A and T cell receptor B loci in families with rheumatoid arthritis. AB - OBJECTIVE: To examine whether the T cell receptor (TCR) A or TCRB loci exhibit linkage with disease in multiplex rheumatoid arthritis (RA) families. METHODS: A linkage study was performed in 184 RA families from the UK Arthritis and Rheumatism Council Repository, each containing at least 1 affected sibpair. The microsatellites D14S50, TCRA, and D14S64 spanning the TCRA locus and D7S509, Vbeta6.7, and D7S688 spanning the TCRB locus were used as DNA markers. The subjects were genotyped using a semiautomated polymerase chain reaction-based method. Two-point and multipoint linkage analyses were performed. RESULTS: Nonparametric single-marker likelihood odds (LOD) scores were 0.49 (P = 0.07) for D14S50, 0.65 (P = 0.04) for TCRA, 0.07 (P = 0.29) for D14S64, 0.01 (P = 0.43) for D7S509, 0.0 (P = 0.50) for Vbeta6.7, and 0.0 (P = 0.50) for D7S688. By multipoint analysis, there was no evidence of linkage at TCRB (LOD score 0), and the maximum LOD score at the TCRA locus was 0.37 (at D14S50). The presence of a susceptibility locus (LOD score < -2.0) was excluded, with lambda > or = 1.8 at TCRA and > or = 1.4 at TCRB. CONCLUSION: These linkage studies provide no significant evidence of a major germline-encoded TCRA or TCRB component of susceptibility to RA. PMID- 9336414 TI - Antibodies against a peptide sequence located in the linker region of the HMG-1/2 box domains in sera from patients with juvenile rheumatoid arthritis. AB - OBJECTIVE: To extend our work on the mapping of B cell epitopes on nucleosomal high mobility group (HMG) proteins in the sera of patients with juvenile rheumatoid arthritis (JRA). METHODS: Seventy-seven pauciarticular-onset JRA serum samples from antinuclear antibody (ANA)-positive patients and 42 polyarticular onset JRA patient sera found to react with HMG-2 by immunoblotting were used in this study. To identify B cell epitopes on HMG-2, recombinant HMG-2 protein fragments were used in enzyme-linked immunosorbent assay (ELISA) and in competition ELISA experiments with a set of overlapping synthetic peptides. Fine epitope mapping was achieved by oligopeptide synthesis, followed by immunoblotting. RESULTS: Pauciarticular, but not polyarticular, JRA patient sera were found to recognize a lysine-rich major epitope (KKGKKKDP), which is located in the linker region of the HMG box domains of the HMG-2 nonhistone chromosomal protein. No significant immunoreactions were observed in sera from ANA-negative JRA patients and in sera from children with nonrheumatic diseases, indicating that this epitope seems to be specific for pauciarticular-onset JRA. CONCLUSION: In addition to our previous finding that JRA sera will react with a defined epitope on HMG-17, pauciarticular JRA patient sera were also found to recognize a defined epitope on the HMG-2 protein, thus suggesting the importance of this epitope in the etiology of JRA. PMID- 9336415 TI - Increased nitric oxide production accompanied by the up-regulation of inducible nitric oxide synthase in vascular endothelium from patients with systemic lupus erythematosus. AB - OBJECTIVE: To investigate whether systemic lupus erythematosus (SLE) is accompanied by increased serum nitrite levels, whether active compared with inactive disease is associated with greater nitric oxide (NO) production, and whether endothelial cells or keratinocytes serve as cellular sources of NO by virtue of their increased expression of either constitutive nitric oxide synthase (cNOS) or inducible NOS (iNOS). METHODS: Fifty-one serum samples (46 from patients with SLE) were analyzed for NO production by measuring nitrite levels in a calorimetric assay. Skin biopsy samples from 21 SLE patients and 11 healthy volunteers were evaluated immunohistochemically, using monoclonal antibodies, for endothelial cell and keratinocyte cNOS and iNOS expression. RESULTS: Serum nitrite levels were significantly elevated in the 46 patients with SLE (mean +/- SEM 37 +/- 6 microM/liter) compared with controls (15 +/- 7 microM/liter; P < 0.01), and were elevated in patients with active SLE compared with those with inactive disease (46 +/- 7 microM/liter versus 30 +/- 7 microM/liter; P < 0.01). Serum nitrite levels correlated with disease activity (r = 0.47, P = 0.04) and with levels of antibodies to double-stranded DNA (r = 0.35, P = 0.02). Endothelial cell expression of iNOS in SLE patients (mean +/- SEM score 1.5 +/- 0.2) was significantly greater compared with controls (0.6 +/- 0.2; P < 0.01), and higher in patients with active disease compared with those with inactive SLE (1.7 +/- 0.2 versus 1.2 +/- 0.2; P < 0.01). Keratinocyte expression of iNOS was also significantly elevated in SLE patients (0.9 +/- 0.1) compared with controls (0.4 +/- 0.1; P < 0.001). With regard to expression of cNOS, there were no differences between patients with active SLE, those with inactive SLE, and normal controls in either the vascular endothelium or the keratinocytes. CONCLUSION: NO production is increased in patients with SLE, and 2 potential sources of excessive NO are activated endothelial cells and keratinocytes via up-regulated iNOS. PMID- 9336416 TI - P-31 magnetic resonance spectroscopy demonstrates unaltered muscle energy utilization in polymyalgia rheumatica. AB - OBJECTIVE: To characterize muscle metabolic anomalies associated with polymyalgia rheumatica (PMR) using P-31 magnetic resonance spectroscopy (MRS). METHODS: Seventeen patients with PMR and 9 age-matched control subjects were investigated. The forearm flexor muscles were examined by P-31 MRS in a 4.7 T superconducting horizontal magnet (Biospec Bruker 47/30) during a rest-exercise-recovery protocol. The intracellular pH and the relative concentrations of phosphocreatine (PCr), inorganic phosphate (Pi), ATP, and phosphomonoesters were measured every minute during the protocol. Based on the PCr and pH time-dependent changes during exercise and recovery, the rates of ATP production from PCr hydrolysis, glycogenolysis, and aerobic metabolism were calculated for each minute of exercise. RESULTS: At rest, the metabolic parameters [PCr]:[Pi], pH, and [PCr]:[ATP] were not significantly different between the PMR patients and the control subjects. During exercise, the energetic cost and the contribution of anaerobic and oxidative pathways to energy supply were similar in the 2 groups, as were the recovery kinetics of the phosphorylated compounds and pH. CONCLUSION: Muscle MRS does not confirm the findings of histologic and biochemical studies that suggest an alteration of mitochondrial function in PMR. No other modifications in glycolytic or glycogenolytic pathways or in proton handling were found that could indicate an alteration of muscle energetics in patients with PMR. PMID- 9336417 TI - Susceptibility to ankylosing spondylitis in twins: the role of genes, HLA, and the environment. AB - OBJECTIVE: To determine the relative effects of genetic and environmental factors in susceptibility to ankylosing spondylitis (AS). METHODS: Twins with AS were identified from the Royal National Hospital for Rheumatic Diseases database. Clinical and radiographic examinations were performed to establish diagnoses, and disease severity was assessed using a combination of validated scoring systems. HLA typing for HLA-B27, HLA-B60, and HLA-DR1 was performed by polymerase chain reaction with sequence-specific primers, and zygosity was assessed using microsatellite markers. Genetic and environmental variance components were assessed with the program Mx, using data from this and previous studies of twins with AS. RESULTS: Six of 8 monozygotic (MZ) twin pairs were disease concordant, compared with 4 of 15 B27-positive dizygotic (DZ) twin pairs (27%) and 4 of 32 DZ twin pairs overall (12.5%). Nonsignificant increases in similarity with regard to age at disease onset and all of the disease severity scores assessed were noted in disease-concordant MZ twins compared with concordant DZ twins. HLA-B27 and B60 were associated with the disease in probands, and the rate of disease concordance was significantly increased among DZ twin pairs in which the co-twin was positive for both B27 and DR1. Additive genetic effects were estimated to contribute 97% of the population variance. CONCLUSION: Susceptibility to AS is largely genetically determined, and the environmental trigger for the disease is probably ubiquitous. HLA-B27 accounts for a minority of the overall genetic susceptibility to AS. PMID- 9336418 TI - Clinical, laboratory, radiographic, and histopathologic features of methotrexate associated lung injury in patients with rheumatoid arthritis: a multicenter study with literature review. AB - OBJECTIVE: To describe the clinical, laboratory, radiologic, and histopathologic features of methotrexate (MTX)-induced lung injury in a combined cohort of selected patients with rheumatoid arthritis (RA) and all cases reported in the English-language literature. METHODS: Retrospective combined cohort review and abstraction from the medical literature. Case reports were obtained from 6 centers that had 4 or more cases of potential MTX lung injury per site. RA patients who were seen between 1981 and 1993 and who satisfied predetermined criteria for the presence of MTX lung injury were identified. RESULTS: Twenty seven patients satisfied the criteria for definite MTX lung injury, and 2 for probable MTX lung injury. Predominant clinical features of MTX lung injury included shortness of breath in 27 patients (93.1%), which was present for 23.5 +/- 22.3 days (mean +/- SD), cough in 24 (82.8%), present for 26.9 +/- 28.5 days, and fever in 20 (69.0%), present for 10.4 +/- 12.8 days. Five patients (17.2%) died, compared with 12 of 68 (17.6%) reported in the medical literature. Four of the 6 patients who were re-treated with MTX after an initial pulmonary event developed recurrent lung toxicity, resulting in 2 deaths, compared with a recurrence rate of 3 of 6 in the literature. CONCLUSION: MTX lung injury is most often a subacute process, in which symptoms are commonly present for several weeks before diagnosis. Approximately 50% of the cases are diagnosed within 32 weeks from initiation of MTX treatment. A patient who recovers from MTX lung injury should not be re-treated. Earlier recognition and drug withdrawal may avoid the serious and sometimes fatal outcome that has been observed in this and other studies. PMID- 9336419 TI - A phase 1 study to address the safety and efficacy of granulocyte colony stimulating factor for the mobilization of hematopoietic progenitor cells in active rheumatoid arthritis. AB - OBJECTIVE: To examine the safety and efficacy of granulocyte colony-stimulating factor (G-CSF) alone for the mobilization of peripheral blood progenitor cells in patients with resistant active rheumatoid arthritis (RA). METHODS: Five patients with resistant active RA were studied. A dose of 5 microg/kg of G-CSF (Filgrastim) was given subcutaneously each day for 5 days, and the number of stem cells mobilized into the peripheral blood was assessed by daily CD34 counts. RA disease activity was assessed by standard clinical methods. RESULTS: The absolute numbers of peripheral blood CD34+ cells peaked on day 4, with a mean value of 0.025 x 10(9)/liter (range 0.013-0.048 x 10(9)/liter). There was no significant change in disease activity during the study or in the month following therapy. CONCLUSION: Using G-CSF alone, CD34+ progenitor peripheral blood cells were mobilized in numbers suitable for leukopheresis. G-CSF therapy was well-tolerated in patients with active RA, and was not associated with a flare during treatment or in the month following treatment. PMID- 9336420 TI - Combination treatment of severe rheumatoid arthritis with cyclosporine and methotrexate for forty-eight weeks: an open-label extension study. The Methotrexate-Cyclosporine Combination Study Group. AB - OBJECTIVE: To determine whether the clinical benefit and favorable safety profile previously noted with the combination of cyclosporine (CSA) and methotrexate (MTX) given for 24 weeks in patients with rheumatoid arthritis (RA) would be maintained for a further 24 weeks, and whether the addition of CSA in patients who had previously been randomized to receive placebo + MTX would result in clinical benefit. METHODS: Eligible subjects from the initial study (weeks 0-24), in which the addition of placebo or CSA to MTX therapy was compared in patients with RA that was partially responsive to MTX, were enrolled. Patients who had received CSA + MTX continued this regimen for a further 24 weeks (weeks 24-48) (group 1; n = 48), and patients who had initially received placebo + MTX now received CSA + MTX for 24 weeks (weeks 24-48) (group 2; n = 44), in an open-label extension study. The primary outcome measures were the number of tender joints, number of swollen joints, physician and patient global assessments, pain, functional disability as measured by the modified Health Assessment Questionnaire, and erythrocyte sedimentation rate. RESULTS: Of the 92 patients enrolled, 80 (87%) completed the extension study. In patients in group 1, the clinically and statistically significant improvement in response outcomes previously noted at week 24, ranging from 25% to 50%, was maintained through week 48. In patients in group 2, the addition of CSA resulted in significant clinical improvement. By week 48, most outcome measures in group 2 patients were similar to those in group 1 patients. CSA treatment resulted in a small increase in serum creatinine levels, but only 1 patient was withdrawn from the study for this reason. CONCLUSION: The clinical improvement previously observed in patients treated with the CSA + MTX combination for 24 weeks was maintained for 24 subsequent weeks, without serious adverse effects, and was also observed in the patients whose treatment was switched from placebo + MTX to CSA + MTX. PMID- 9336421 TI - Trial of intravenous pulse cyclophosphamide and methylprednisolone in the treatment of severe systemic-onset juvenile rheumatoid arthritis. AB - OBJECTIVE: Not uncommonly, some children with systemic-onset juvenile rheumatoid arthritis (JRA) have persistently active disease with joint destruction and profound growth delay despite maximum treatment with known medications. Based on previous observations of improvement in synovitis following intravenous (I.V.) cyclophosphamide (CYC) and methylprednisolone (MP) treatments, a group of children with severe systemic-onset JRA was treated in an attempt to control active synovitis and to allow tapering of corticosteroids. METHODS: Four patients with systemic-onset JRA were continued on a daily regimen of nonsteroidal antiinflammatory agents and prednisone, with a weekly subcutaneous dose of methotrexate (1 mg/kg). In addition, 1 patient continued receiving sulfasalazine and 1 patient remained on a regimen of sulfasalazine and hydroxychloroquine. Patients received 6-10 monthly treatments of I.V. CYC (500-1,000 mg/m2) and MP (30 mg/kg; 1 gm maximum) accompanied by I.V. mesna and large amounts of I.V. fluids. Subsequent treatments were given once every 2-3 months. RESULTS: After 12 20 I.V. pulses of CYC, all patients showed improvement, and 3 achieved remission of disease. All were able to discontinue corticosteroid use and all had an increase in linear growth. CONCLUSION: Monthly I.V. pulse CYC treatments can be useful to control disease in selected children with severe, destructive JRA. PMID- 9336422 TI - Magnetic resonance imaging-determined synovial membrane and joint effusion volumes in rheumatoid arthritis and osteoarthritis: comparison with the macroscopic and microscopic appearance of the synovium. AB - OBJECTIVE: To evaluate the relationship between synovial membrane and joint effusion volumes determined by magnetic resonance imaging (MRI) and macroscopic and microscopic synovial pathologic findings in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Synovial biopsies were performed, and macroscopic grades of synovitis assigned, at preselected knee sites during arthroscopy or arthrotomy in 17 knees with RA and 25 with OA. Synovial inflammation and 9 separate tissue characteristics were graded histologically. Synovial membrane and joint effusion volumes were determined by preoperative MRI, enhanced with intravenous gadopentetate dimeglumine. RESULTS: MRI-determined synovial membrane volumes were correlated with the overall histologic assessment of synovial inflammation (Spearman's sigma = 0.55, P < 0.001), with fibrin deposition, with subsynovial mononuclear and polymorphonuclear leukocyte infiltration (sigma = 0.51-0.59), and less significantly with macroscopic synovitis, vessel proliferation, and granulation tissue formation (sigma = 0.40-0.42). No correlation with synovial lining multiplication, perivascular edema, villous formation, or fibrosis was found (sigma < 0.30). CONCLUSION: MRI-determined synovial volumes are correlated with synovial inflammatory activity. Synovial volumes probably mainly reflect the mass of cell-infiltrated, vascularized subsynovial tissue, but may also be influenced by the cumulative synovial proliferative activity. MRI-determined synovial membrane and effusion volumes may be sensitive markers and/or predictors of disease activity and treatment outcome in RA. PMID- 9336423 TI - Mortality studies in psoriatic arthritis: results from a single outpatient clinic. I. Causes and risk of death. AB - OBJECTIVE: To identify the causes of death and mortality risk in patients with psoriatic arthritis (PsA) who were being followed up at a single outpatient clinic in Toronto, Ontario, Canada. METHODS: Patients enrolled in the PsA Clinic between 1978 and 1993 were compared with the general population of Ontario. Deaths were identified from the clinic database and through linkage with the provincial mortality database, and causes were confirmed by death certificates. A standardized mortality ratio (SMR) was computed, based on the assumption that patients lost to followup were alive at the end of the study. RESULTS: Of the 428 patients with PsA (194 women and 234 men), 53 (26 women and 27 men) died. The 4 leading causes of death were diseases of the circulatory (36.2%) or respiratory (21.3%) system, malignant neoplasms (17.0%), and injuries/poisoning (14.9%). The SMR for the female cohort was 1.59, and for the men, it was 1.65, indicating a 59% and 65% increase in the death rate, respectively. Deaths due to respiratory causes were particularly increased in these patients. CONCLUSION: The results suggest that this PsA Clinic outpatient population had an increased mortality risk. PMID- 9336424 TI - Adverse outcomes of antiinflammatory therapy among patients with polymyalgia rheumatica. AB - OBJECTIVE: To evaluate the incidence and risks of adverse events associated with therapy (both corticosteroids [CS] and nonsteroidal antiinflammatory drugs [NSAIDs]) among a previously identified, population-based cohort of patients first diagnosed with polymyalgia rheumatica (PMR) between 1970 and 1991 who were followed up over the long term. METHODS: Information on demographics, PMR diagnosis, disease course, and drug therapy, in addition to data on adverse events commonly associated with CS and NSAID treatment, was obtained from the Rochester Epidemiology Project database. Cox proportional hazards and regression analysis models were used to evaluate the relationship between the occurrence of these events and therapy. RESULTS: Of the 232 patients (69 male, 163 female) included in the study, the mean age at PMR diagnosis was 72.9 years, the average followup was 8.0 years, and 30 patients were also diagnosed with giant cell (temporal) arteritis. Among the 175 patients (49 male, 126 female) treated with CS, the mean duration of CS therapy was 2.4 years, the average daily dose was 9.6 mg, and the mean cumulative dose was 8.4 gm. In total, 65% of the 124 patients treated with CS alone experienced at least 1 adverse event, compared with 67% of the 57 patients treated with NSAIDs alone and 80% of the 51 patients treated with CS and NSAIDs. The average time from initiation of therapy to the first adverse event was 1.6 years (n = 160). Proportional hazards modeling identified 3 variables that independently increased the risk of adverse events: age at PMR diagnosis, a cumulative dose of prednisone > or = 1,800 mg, and female sex. Person-year analysis revealed that the risks of diabetes mellitus, vertebral fractures, femoral neck fractures, and hip fractures were 2-5 times greater among PMR patients compared with age- and sex-matched individuals from the same population. Medical care or consultation by a rheumatologist was a highly significant predictor of a lower initial CS dose. CONCLUSION: The use of CS and NSAIDs in the treatment of PMR is associated with important long-term morbidity. PMID- 9336425 TI - Criteria for the diagnosis of familial Mediterranean fever. AB - OBJECTIVE: To establish a new set of criteria for the diagnosis of familial Mediterranean fever (FMF). METHODS: Twenty-seven features and manifestations typical of FMF were studied to determine their prevalence in 105 patients with FMF and 106 controls. Diagnosis of FMF in the study group was based on clinical judgment. Controls were patients with a variety of other diseases who presented to the emergency room or outpatient clinics with recurrent episodes of pain in body sites usually involved in FMF attacks. Manifestations observed to be significantly more common in FMF patients than in controls were incorporated into the rule proposed for diagnosis of FMF, based on a model of major, minor, and supportive criteria. RESULTS: Two sets of diagnostic criteria were established. A conservative criteria set for diagnosis of FMF was based on the presence of 1 major or 2 minor criteria, or 1 minor plus 5 supportive criteria, and a simple criteria set for diagnosis of FMF required 1 major or 2 minor criteria. The sensitivity and specificity of these 2 criteria sets were >95% and >97%, respectively. CONCLUSION: The proposed new sets of criteria were highly sensitive and specific, and could be used to readily diagnose FMF and to distinguish FMF from other periodic febrile diseases. PMID- 9336426 TI - Percutaneous needle muscle biopsy in the evaluation of patients with suspected inflammatory myopathy. AB - OBJECTIVE: To determine the usefulness of a unique method of percutaneous needle muscle biopsy (NMB) in patients with suspected idiopathic inflammatory myopathy (IIM). METHODS: The yield of percutaneous NMB was studied in 55 patients who were found to have a combination of clinical, laboratory, or electromyographic features of IIM. RESULTS: A diagnosis of IIM was confirmed histopathologically in 29 patients (53%), other specific myopathies were found in 5 (9%), nonspecific myopathic changes were present in 11 (20%), and a neurogenic process was diagnosed in 3 (5%). Nonspecific changes or no abnormalities were present in 7 patients (13%). Followup of the 18 patients with nonspecific histopathologic findings disclosed that only 3 had a subsequent disease course compatible with IIM. CONCLUSION: Percutaneous NMB is a safe, convenient, and relatively inexpensive method of muscle biopsy, with a high diagnostic yield for the pathologic confirmation of IIM. It should be considered as a primary method of acquiring muscle for histopathologic examination in the evaluation of suspected IIM. PMID- 9336427 TI - Symptomatic spinal calcinosis in systemic sclerosis (scleroderma). AB - Two patients with diffuse cutaneous systemic sclerosis and spinal calcification, involving the lumbar spine in one and the cervical spine in the other, are described. Computed tomography-guided aspiration of the calcific masses was performed, and material aspirated from one patient was shown to be apatite, Ca5(PO4)3OH. One patient showed improvement following lumbar laminotomy, hemilaminectomy, and diskectomy. PMID- 9336428 TI - Azathioprine-induced severe pancytopenia due to a homozygous two-point mutation of the thiopurine methyltransferase gene in a patient with juvenile HLA-B27 associated spondylarthritis. AB - Severe pancytopenia due to azathioprine (AZA) toxicity in patients with autoimmune diseases is not uncommon. We describe a 14-year-old girl with HLA-B27+ spondylarthritis who was treated with AZA 3 mg/kg/day and who suddenly developed severe pancytopenia in the seventh week of treatment. Analysis of the catabolic pathway of AZA revealed a homozygous deficiency of thiopurine methyltransferase (TPMT) on the basis of a combined 2-point mutation at nucleotide positions 460 and 719 in the gene for TPMT, causing a toxic level of the metabolic active 6 thioguanine nucleotides (6-TGN) (2,394 pmoles/8 x 10(8) red blood cells). The patient was transfusion dependent and finally recovered 8 weeks after the development of the pancytopenia. At that time, 6-TGN had already returned to normal therapeutic levels. Family studies revealed another homozygous deficiency in the mother, while the other family members were heterozygous. PMID- 9336429 TI - Systemic lupus erythematosus with membranous glomerulonephritis and transverse myelitis associated with anabolic steroid use. AB - This report describes a 29-year-old bodybuilder taking anabolic steroids who presented with urinary retention, arthralgias, and peripheral edema, subsequently developed acute lower-extremity paralysis, and was diagnosed as having transverse myelitis and membranous glomerulonephritis secondary to systemic lupus erythematosus (SLE). The association of anabolic steroid use and hyperprolactinemia, and their possible link to the development of SLE, are reviewed. PMID- 9336430 TI - Amyloid arthropathy. PMID- 9336431 TI - Lack of serologic evidence for involvement of human herpesvirus 8 in autoimmune diseases. PMID- 9336432 TI - Familial antiphospholipid syndrome and HLA-DRB gene associations. PMID- 9336433 TI - In defense of practice guidelines in rheumatology: comment on the article by Cohen. PMID- 9336434 TI - Development of guidelines as educational tools: comment on the article by Cohen. PMID- 9336435 TI - "Expert opinion" versus hard science: comment on the review by Wallace et al. PMID- 9336436 TI - Sulfasalazine and human immunodeficiency virus-associated reactive arthritis: comment on the article by Clegg et al. PMID- 9336437 TI - Comparison of sulfasalazine and placebo in the treatment of ankylosing spondylitis: comment on the article by Clegg et al. PMID- 9336438 TI - Expression of apoptosis-related Fas antigen and in vitro apoptosis of lymphocyte subsets from patients with primary Sjogren's syndrome: comment on the article by Lorenz et al. PMID- 9336439 TI - Is one year early, or too late? Comment on the article by Tak et al. PMID- 9336440 TI - Local steroid injections: comment on the American College of Rheumatology guidelines for the management of osteoarthritis of the hip and on the letter by Swezey. PMID- 9336441 TI - The prethrombotic/hypercoagulable state of Behcet's disease: comment on the article by Golden et al. PMID- 9336442 TI - Prelingual deafness: high prevalence of a 30delG mutation in the connexin 26 gene. AB - Prelingual non-syndromic (isolated) deafness is the most frequent hereditary sensory defect. In >80% of the cases, the mode of transmission is autosomal recessive. To date, 14 loci have been identified for the recessive forms (DFNB loci). For two of them, DFNB1 and DFNB2, the genes responsible have been characterized; they encode connexin 26 and myosin VIIA, respectively. In order to evaluate the extent to which the connexin 26 gene (Cx26) contributes to prelingual deafness, we searched for mutations in this gene in 65 affected Caucasian families originating from various countries, mainly tunisia, France, New Zealand and the UK. Six of these families are consanguineous, and deafness was shown to be linked to the DFNB1 locus, 10 are small non consanguineous families in which the segregation of the trait has been found to be compatible with the involvement of DFNB1, and in the remaining 49 families no linkage analysis has been performed. A total of 62 mutant alleles in 39 families were identified. Therefore, mutations in Cx26 represent a major cause of recessively inherited prelingual deafness since according to the present results they would underlie approximately half of the cases. In addition, one specific mutation, 30delG, accounts for the majority (approximately 70%) of the Cx26 mutant alleles. It is therefore one of the most frequent disease mutations so far identified. Several lines of evidence indicate that the high prevalence of the 30delG mutation arises from a mutation hot spot rather than from a founder effect. Genetic counseling for prelingual deafness has been so far considerably impaired by the difficulty in distinguishing genetic and non genetic deafness in families presenting with a single deaf child. Based on the results presented here, the development of a simple molecular test could be designed which should be of considerable help. PMID- 9336443 TI - Analysis of TALE superclass homeobox genes (MEIS, PBC, KNOX, Iroquois, TGIF) reveals a novel domain conserved between plants and animals. AB - A new Caenorhabditis elegans homeobox gene, ceh-25, is described that belongs to the TALE superclass of atypical homeodomains, which are characterized by three extra residues between helix 1 and helix 2. ORF and PCR analysis revealed a novel type of alternative splicing within the homeobox. The alternative splicing occurs such that two different homeodomains can be generated, which differ in their first 25 amino acids. ceh-25 is an orthologue of the vertebrate Meis genes and it shares a new conserved domain of 130 amino acids with them. A thorough analysis of all TALE homeobox genes was performed and a new classification is presented. Four TALE classes are identified in animals: PBC, MEIS, TGIF and IRO (Iroquois); two types in fungi: the mating type genes (M-ATYP) and the CUP genes; and two types in plants: KNOX and BEL. The IRO class has a new conserved motif downstream of the homeodomain. For the KNOX class, a conserved domain, the KNOX domain, was defined upstream of the homeodomain. Comparison of the KNOX domain and the MEIS domain shows significant sequence similarity revealing the existence of an archetypal group of homeobox genes that encode two associated conserved domains. Thus TALE homeobox genes were already present in the common ancestor of plants, fungi and animals and represent a branch distinct from the typical homeobox genes. PMID- 9336444 TI - Processing of topoisomerase I cleavable complexes into DNA damage by transcription. AB - Topoisomerase I (TOP1)-mediated DNA damage induced by camptothecin (CPT) in the presence of active transcription has been studied using purified calf thymus TOP1 and T7 RNA polymerase. CPT-stabilized TOP1 cleavable complexes located on the template strand within the transcribed region were found to be converted into irreversible strand breaks by the elongating RNA polymerase. By contrast, CPT stabilized TOP1 cleavable complexes located on the non-template strand within the transcribed region was unaffected by the elongating RNA polymerase. Previous studies have demonstrated that the elongating T7 RNA polymerase is arrested by TOP1 cleavable complexes located on the template but not the non-template strand [Bendixen et al ., (1990) Biochemistry , 29, 5613-5619]. Together, these results suggest a model in which collision between the TOP1-cleavable complexes located on the template strand and the elongating RNA polymerase results in transcription arrest and conversion of TOP1 cleavable complexes into 'irreversible' strand breaks. The implication of the transcription collision model in DNA damage and repair, as well as cell killing, is discussed. PMID- 9336445 TI - Triplex formation at physiological pH: comparative studies on DNA triplexes containing 5-Me-dC tethered at N4 with spermine and tetraethyleneoxyamine. AB - Oligodeoxynucleotides with spermine conjugation at C4 of 5-Me-dC ( sp -ODN) exhibit triple helix formation with complementary Watson-Crick duplexes, and were optimally stable at physiological pH 7.3 and low salt concentration. This was attributed to a favored reassociation of the polycationic third strand with the anionic DNA duplex. To gain further insights into the factors that contribute to the enhancement of triplex stability and for engineering improved triplex systems, the spermine appendage at C4 of 5-Me-dC was replaced with 1,11-diamino 3,6,9-trioxaundecane to create teg -ODNs. From the triple helix forming abilities of these modified ODNs studied by hysteresis behaviour and the effect of salts on triplex stability, it is demonstrated here that teg- ODNs stabilise triplexes through hydrophobic desolvation while sp -ODNs stabilise triplexes by charge effects. The results imply that factors in addition to base stacking effects and interstrand hydrogen bonds are significantly involved in modulation of triplex stability by base modified oligonucleotides. PMID- 9336446 TI - The tRNATyr-isoacceptors and their genes in the ciliate Tetrahymena thermophila: cytoplasmic tRNATyr has a QPsiA anticodon and is coded by multiple intron containing genes. AB - In the ciliated protozoa Tetrahymena thermophila introns have been detected in rRNA and mRNAs until now. We have isolated and sequenced seven tRNATyr genes from the T.thermophila nuclear genome. All of these genes contain introns of identical length and sequence. The 11 bp long intervening sequences are located 1 nt 3' to the anticodon as found in other eukaryotic nuclear tRNA genes. Tetrahymena tRNATyr genes are efficiently transcribed in HeLa cell nuclear extract. Moreover, processing and splicing occurred in HeLa as well as in wheat germ extracts, supporting the notion that Tetrahymena tRNATyr introns can be classified as authentic tRNA introns. We have also isolated cytoplasmic tRNATyr from Tetrahymena cells. This tRNATyr isoacceptor has a QPsiA anticodon and is not a UAG suppressor as shown in in vitro translation studies. Since UAG and UAA codons are used as glutamine codons in Tetrahymena macronuclear DNA, the presence of a strong natural UAG suppressor such as tRNATyr with GPsiA anticodon should cause misreading of the glutamine as tyrosine codons and the absence of the latter had thus been predicted. Furthermore we have studied the organization of tRNATyr genes in the genome of T.thermophila and have found two types of tRNATyr gene arrangement. A minimum of 12 tRNATyr genes are present as single copies in genomic DNA HindIII restriction fragments ranging in size from 0.6 to 7 kb. Additionally one cluster of tRNATyr genes consisting of six members has been detected in a 2.3 kb HindIII fragment. PMID- 9336447 TI - Rescue of the RNA phage genome from RNase III cleavage. AB - The secondary structure of the RNA from the single-stranded RNA bacteriophages, like MS2 and Qb, has evolved to serve a variety of functions such as controlling gene expression, exposing binding sites for the replicase and capsid proteins, allowing strand separation and so forth. On the other hand, all of these foldings have to perform in bacterial cells in which various RNA splitting enzymes are present. We therefore examined whether phage RNA structure is under selective pressure by host RNases. Here we show this to be true for RNase III. A fully double-stranded hairpin of 17 bp, which is an RNase III target, was inserted into a non-coding region of the MS2 RNA genome. In an RNase III-host these phages survived but in wild-type bacteria they did not. Here the stem underwent Darwinian evolution to a structure that was no longer a substrate for RNase III. This was achieved in three different ways: (i) the perfect stem was maintained but shortened by removing all or most of the insert; (ii) the stem acquired suppressor mutations that replaced Watson-Crick base pairs by mismatches; (iii) the stem acquired small deletions or insertions that created bulges. These insertions consist of short stretches of non-templated A or U residues. Their origin is ascribed to polyadenylation at the site of the RNase III cut (in the + or - strand) either by Escherichia coli poly(A) polymerase or by idling MS2 replicase. PMID- 9336449 TI - Method for phosphorothioate antisense DNA sequencing by capillary electrophoresis with UV detection. AB - The progress of antisense DNA therapy demands development of reliable and convenient methods for sequencing short single-stranded oligonucleotides. A method of phosphorothioate antisense DNA sequencing analysis using UV detection coupled to capillary electrophoresis (CE) has been developed based on a modified chain termination sequencing method. The proposed method reduces the sequencing cost since it uses affordable CE-UV instrumentation and requires no labeling with minimal sample processing before analysis. Cycle sequencing with ThermoSequenase generates quantities of sequencing products that are readily detectable by UV. Discrimination of undesired components from sequencing products in the reaction mixture, previously accomplished by fluorescent or radioactive labeling, is now achieved by bringing concentrations of undesired components below the UV detection range which yields a 'clean', well defined sequence. UV detection coupled with CE offers additional conveniences for sequencing since it can be accomplished with commercially available CE-UV equipment and is readily amenable to automation. PMID- 9336448 TI - An efficient protocol for linker scanning mutagenesis: analysis of the translational regulation of an Escherichia coli RNA polymerase subunit gene. AB - A protocol has been developed that is capable of saturating regions hundreds of basepairs in length with linker scanning mutations. The efficacy of this method stems from the design of the linker scanning mutagenesis (LSM) cassette which is composed of a selectable marker flanked by two oligonucleotides, each of which contains a recognition site for a different restriction endonuclease. The cleavage site for one endonuclease is within its recognition site, while the second endonuclease cleaves in the target DNA beyond the end of the cassette. Digestion with these endonucleases and subsequent ligation results in the replacement of 12 bp of the original target sequence with 12 bp of the linker scanning oligonucleotide. We have used this protocol to mutagenize a span of approximately 400 bp surrounding the start site of the gene for the beta subunit (rpoB) of Escherichia coli RNA polymerase. The translation of the beta mRNA has been shown previously to be regulated by the intracellular concentration of either beta or beta'. Analysis of the linker scanning mutations indicates that sequences extending a considerable distance both upstream and downstream of the start site are required for normal translation. Also a site that appears to be involved in translational repression by excess beta' has been identified. PMID- 9336450 TI - Prokaryotic 5'-3' exonucleases share a common core structure with gamma-delta resolvase. AB - The three dimensional crystal structure of T5 5'-3' exonuclease was compared with that of two other members of the 5'-3' exonuclease family: T4 ribonuclease H and the N-terminal domain of Thermus aquaticus DNA polymerase I. Though these structures were largely similar, some regions of these enzymes show evidence of significant molecular flexibility. Previous sequence analysis had suggested the existence of a helix-hairpin-helix motif in T5 exonuclease, but a distinct, though related structure is actually found to occur. The entire T5 exonuclease structure was then compared with all the structures in the complete Protein Data Bank and an unexpected similarity with gamma-delta (gamma delta) resolvase was observed. 5'-3' exonucleases and gamma delta resolvase are enzymes involved in carrying out quite different manipulations on nucleic acids. They appear to be unrelated at the primary sequence level, yet the fold of the entire catalytic domain of gamma delta resolvase is contained within that of the 5'-3'exonuclease. Different large-scale helical structures are used by both families to form DNA binding sites. PMID- 9336451 TI - Evidence that Snf-Swi controls chromatin structure over both the TATA and UAS regions of the SUC2 promoter in Saccharomyces cerevisiae. AB - The Snf-Swi complex of the yeast Saccharomyces cerevisiae has been shown to control gene expression by controlling chromatin structure. We have analyzed the promoter of the SUC2 gene, a gene strongly controlled by Snf-Swi, by a high resolution analysis of micrococcal nuclease digests. This analysis suggests that there are at least four nucleosomes positioned over the SUC2 TATA and UAS regions under conditions repressing SUC2 transcription. Under derepressing conditions this entire promoter region is much more sensitive to MNase digestion. Analysis of an snf2 Delta mutant demonstrates that even under derepressing conditions the SUC2 promoter is resistant to MNase digestion. Thus, the Snf-Swi complex appears to control chromatin structure over both the SUC2 TATA and UAS regions. The presence of nucleosomes over both promoter regions may explain the strong requirement of SUC2 for Snf-Swi function. PMID- 9336452 TI - Molecular cloning of four novel murine ribonuclease genes: unusual expansion within the ribonuclease A gene family. AB - We have characterized four novel murine ribonuclease genes that, together with the murine eosinophil-associated ribonucleases 1 and 2, form a distinct and unusual cluster within the RNase A gene superfamily. Three of these genes (mR-3, mR-4, mR-5) include complete open reading frames, encoding ribonucleases with eight cysteines and appropriately spaced histidines (His11 and His124) and lysine (Lys35) that are characteristic of this enlarging protein family; the fourth sequence encodes a non-functional pseudogene (mR-6P). Although the amino acid sequence similarities among these murine ribonucleases varies from 60 to 94%, they form a unique cluster, as each sequence is found to be more closely related to another of this group than to either murine angiogenin or to murine pancreatic ribonuclease. Interestingly, the relationship between the six genes in this 'mR cluster' and the defined lineages of the RNase A gene family could not be determined by amino acid sequence homology, suggesting the possibility that there are one or more additional ribonuclease lineages that have yet to be defined. Although the nature of the evolutionary constraints promoting this unusual expansion and diversification remain unclear, the implications with respect to function are intriguing. PMID- 9336453 TI - Asymmetry in Flp-mediated cleavage. AB - Flp is a member of the integrase family of site-specific recombinases. Members of the integrase family mediate DNA strand cleavage via a transesterification reaction involving an active site tyrosine residue. The first step of the reaction results in covalent linkage of the protein to the 3'-phosphoryl DNA terminus, leaving a 5'-hydroxyl group at the site of the nick. We have used Flp recognition target (FRT) sites containing a 5'-bridging phosphorothioate linkage at the site of Flp cleavage to accumulate intermediates in which Flp is covalently bound at a cleavage site. We have probed these intermediates with dimethylsulfate using methylation protection and find that Flp-mediated cleavage is associated with protection of two adenine residues that are opposite the sites of cleavage and covalent attachment by Flp. Methylation interference studies showed that cleavage and covalent attachment are also accompanied by differences in the contacts of Flp with each of the two cleavage sites and with the surrounding symmetry elements. Therefore, we provide evidence that Flp-mediated cleavage and covalent attachment result in changes to the conformation of the Flp FRT complex. These changes may be required for Flp-mediated strand exchange activity. PMID- 9336454 TI - An Abf1p C-terminal region lacking transcriptional activation potential stimulates a yeast origin of replication. AB - Although it has been demonstrated that eukaryotic cellular origins of DNA replication may harbor stimulatory elements that bind transcription factors, how these factors stimulate origin function is unknown. In Saccharomyces cerevisiae , the transcription factor Abf1p stimulates origin function of ARS121 and ARS1 . In the results presented here, an analysis of Abf1p function has been carried out utilizing LexA(BD)-Abf1p fusion proteins and an ARS 121 derivative harboring LexA DNA-binding sites. A minimal region which stimulates origin function mapped to 50 amino acids within the C-terminus of Abf1p. When tested for transcriptional activation of a LacZ reporter gene, the same LexA(BD)-Abf1p fusion protein had negligible transcriptional activation potential. Therefore, stimulation of ARS 121 may occur independently of a transcriptional activation domain. It has been previously observed that the Gal4p, Rap1p DNA-binding sites and the LexA-Gal4p fusion protein can replace the role of Abf1p in stimulating ARS 1 . Here we show that the stimulatory function of Abf1p at ARS 121 cannot be replaced by these alternative DNA-binding sites and the potent chimeric transcriptional activator LexA(BD)-Gal4(AD)p . Hence, these results strongly suggest that the Abf1p stimulation of replication may differ for ARS 121 and ARS 1 , and imply specificity in the Abf1p/ARS 121 relationship. PMID- 9336455 TI - Recovery of RNA polymerase II synthesis following DNA damage in mutants of Saccharomyces cerevisiae defective in nucleotide excision repair. AB - We have measured the kinetics of the recovery of mRNA synthesis in the inducible GAL10 and RNR3 genes after exposure of yeast cells to ultraviolet (UV) radiation. Such recovery is abolished in mutant strains defective in nucleotide excision repair (NER) of DNA, including a rad23 mutant. Mutants defective in the RAD7 or RAD16 genes, which are required for the repair of the non-transcribed strand but not the transcribed strand of transcriptionally active genes, show slightly faster recovery of RNA synthesis than wild-type strains. A strain deleted of the RAD26 gene, which is known to be required for strand-specific NER in yeast, manifested delayed recovery of mRNA synthesis, whereas a rad28 mutant, which does not show defective strand-specific repair, showed normal kinetics of recovery. Measurement of the recovery of expression of selected individual yeast genes by Northern analysis following exposure of cells to UV radiation apparently correlates directly with the capacity of cells for strand-specific NER. PMID- 9336456 TI - Extension of the range of DNA sequences available for triple helix formation: stabilization of mismatched triplexes by acridine-containing oligonucleotides. AB - Triple helix formation usually requires an oligopyrimidine*oligopurine sequence in the target DNA. A triple helix is destabilized when the oligopyrimidine*oligopurine target contains one (or two) purine*pyrimidine base pair inversion(s). Such an imperfect target sequence can be recognized by a third strand oligonucleotide containing an internally incorporated acridine intercalator facing the inverted purine*pyrimidine base pair(s). The loss of triplex stability due to the mismatch is partially overcome. The stability of triplexes formed at perfect and imperfect target sequences was investigated by UV thermal denaturation experiments. The stabilization provided by an internally incorporated acridine third strand oligonucleotide depends on the sequences flanking the inverted base pair. For triplexes containing a single mismatch the highest stabilization is observed for an acridine or a propanediol tethered to an acridine on its 3'-side facing an inverted A*T base pair and for a cytosine with an acridine incorporated to its 3'-side or a guanine with an acridine at its 5' side facing an inverted G*C base pair. Fluorescence studies provided evidence that the acridine was intercalated into the triplex. The target sequences containing a double base pair inversion which form very unstable triplexes can still be recognized by oligonucleotides provided they contain an appropriately incorporated acridine facing the double mismatch sites. Selectivity for an A*T base pair inversion was observed with an oligonucleotide containing an acridine incorporated at the mismatched site when this site is flanked by two T*A*T base triplets. These results show that the range of DNA base sequences available for triplex formation can be extended by using oligonucleotide intercalator conjugates. PMID- 9336457 TI - The N-terminal half of the influenza virus NS1 protein is sufficient for nuclear retention of mRNA and enhancement of viral mRNA translation. AB - A collection of C-terminal deletion mutants of the influenza A virus NS1 gene has been used to define the regions of the NS1 protein involved in its functionality. Immunofluorescence analyses showed that the NS1 protein sequences downstream from position 81 are not required for nuclear transport. The capacity of these mutants to bind RNA was studied by in vitro binding tests using a model vRNA probe. These experiments showed that the N-terminal 81 amino acids of NS1 protein are sufficient for RNA binding activity. The collection of mutants also served to map the NS1 sequences required for nuclear retention of mRNA and for stimulation of viral mRNA translation, using the NP gene as reporter. The results obtained indicated that the N-terminal 113 amino acids of NS1 protein are sufficient for nuclear retention of mRNA and stimulation of viral mRNA translation. The possibility that this region of the protein may be sufficient for virus viability is discussed in relation to the sequences of NS1 genes of field isolates and to the phenotype of known viral mutants affected in the NS1 gene. PMID- 9336458 TI - Parameters controlling the rate of gene targeting frequency in the protozoan parasite Leishmania. AB - In this study we investigated the role of several parameters governing the efficiency of gene targeting mediated by homologous recombination in the protozoan parasite Leishmania. We evaluated the relative targeting frequencies of different replacement vectors designed to target several sequences within the parasite genome. We found that a decrease in the length of homologous sequences <1 kb on one arm of the vector linearly influences the targeting frequency. No homologous recombination was detected, however, when the flanking homologous regions were <180 bp. A requirement for a very high degree of homology between donor and target sequences was found necessary for efficient gene targeting in Leishmania , as targeted recombination was strongly affected by base pair mismatches. Targeting frequency increased proportionally with copy number of the target only when the target was part of a linear amplicon, but remained unchanged when it was present on circles. Different chromosomal locations were found to be targeted with significantly variable levels of efficiency. Finally, different strains of the same species showed differences in gene targeting frequency. Overall, gene targeting mediated by homologous recombination in Leishmania shares similarities to both the yeast and the mammalian recombination systems. PMID- 9336459 TI - The expression of a novel, epithelium-specific ets transcription factor is restricted to the most differentiated layers in the epidermis. AB - Ets proteins have been implicated in the regulation of gene expression during a variety of biological processes, including growth control, differentiation, development and transformation. More than 35 related proteins containing the 'ets domain' have now been found which specifically interact with DNA sequences encompassing the core tetranucleotide GGAA. Although ets responsive genes have been identified in the epidermis, little is known about their distribution and function in this tissue. We have now demonstrated that epidermis and cultured epidermal keratinocytes synthesize numerous ets proteins. The expression of some of these proteins is regulated as a function of differentiation. Among these is a novel ets transcription factor with a dual DNA-binding specificity, which we have called jen. The expression of jen is not only epithelial specific, but it is the only ets protein so far described, and one of the very few transcription factors whose expression is restricted to the most differentiated epidermal layers. We show that two epidermal marker genes whose expression coincides with that of jen are transregulated by this protein in a complex mode which involves interactions with other transcriptional regulators such as Sp1 and AP1. PMID- 9336461 TI - The efficiency of a cis-cleaving ribozyme in an mRNA coding region is influenced by the translating ribosome in vivo. AB - A cis -cleaving hammerhead ribozyme (Rz) expression system (3A'-Rz) in Escherichia coli has been constructed that can be used to study the involvement of factors that affect ribozyme cleavage in vivo . The ribozyme sequence is placed in the coding region of 3A' mRNA, which is expressed from a semi-synthetic translation assay gene. The size and the 5'-end sequences of the 3' cleavage fragments were determined and the efficiencies of different Rz variants were measured by quantitative primer extension. It is shown that one of the semi active constructs (3A'-RzIII) can be used as an indicator for ribosomes that read through or terminate at a stop codon upstream of the Rz hammerhead sequence in the mRNA. Readthrough of the stop codon in an uncleaved mRNA gives a full length 3A' protein. Termination at the stop codon upstream of the ribozyme sequence gives a shortened termination product. However, the mRNA fragment that should arise as a result of the auto-cleavage does not give rise to any detectable corresponding truncated protein. Besides studies on translating ribosomes, the 3A'-Rz system can be used to isolate mutant strains that are changed in ribozyme activity either from internal base alterations, or changed interacting host factors. PMID- 9336460 TI - Expression of the thyroid hormone receptor gene, erbAalpha, in B lymphocytes: alternative mRNA processing is independent of differentiation but correlates with antisense RNA levels. AB - The erbAalpha gene encodes two alpha-thyroid hormone receptor isoforms, TRalpha1 and TRalpha2, which arise from alternatively processed mRNAs, erbAalpha1 (alpha1) and erb alpha2 (alpha2). The splicing and alternative polyadenylation patterns of these mRNAs resemble that of mRNAs encoding different forms of immunoglobulin heavy chains, which are regulated at the level of alternative processing during B cell differentiation. This study examines the levels of erbAalpha mRNA in eight B cell lines representing four stages of differentiation in order to determine whether regulation of the alternatively processed alpha1 and alpha2 mRNAs parallels the processing of immunoglobulin heavy chain mRNAs. Results show that the pattern of alpha1 and alpha2 mRNA expression is clearly different from that observed for immunoglobulin heavy chain mRNAs. B cell lines display characteristic ratios of alpha1/alpha2 mRNA at distinct stages of differentiation. Furthermore, expression of an overlapping gene, Rev-ErbAalpha (RevErb), was found to correlate strongly with an increase in the ratio of alpha1/alpha2 mRNA. These results suggest that alternative processing of erbAalpha mRNAs is regulated by a mechanism which is distinct from that regulating immunoglobulin mRNA. The correlation between RevErb and erbAalpha mRNA is consistent with negative regulation of alpha2 via antisense interactions with the complementary RevErb mRNA. PMID- 9336462 TI - Allosteric interaction of the 1alpha,25-dihydroxyvitamin D3 receptor and the retinoid X receptor on DNA. AB - Genomic actions of the hormone 1alpha,25-dihydroxy-vitamin D3(VD) are mediated by the transcription factor VDR, which is a member of the nuclear receptor superfamily. VDR acts in most cases as a heterodimeric complex with the retinoid X receptor (RXR) from specific DNA sequences in the promoter of VD target genes called VD response elements (VDREs). This study describes a mutation (K45A) of the VDR DNA binding domain that enhances the affinity and ligand responsiveness of VDR-RXR heterodimers on some VDREs. In analogy to a homologous mutation in the glucocorticoid receptor (K461A), this lysine residue appears to function as an allosteric 'lock'. Interestingly, overexpression of RXR was found to reduce the responsiveness and sensitivity of wild type VDR to VD, but enhance the response of VDRK45A. Moreover, the transactivation domains of both VDR and RXR were shown to be essential for obtaining responsiveness of the heterodimers to VD and 9- cis retinoic acid (the RXR ligand). This indicates that RXR is an active rather than silent partner of the VDR on the VDREs tested. Taken together, transactivation by VDR-RXR heterodimers can be triggered individually by all components of the protein-DNA complex, but full potency appears to be reached through allosteric interaction. PMID- 9336465 TI - Identification and characterization of a telomerase activity from Schizosaccharomyces pombe. AB - A telomerase-like primer extension activity has been detected in chromatographic fractions derived from Schizosaccharomyces pombe extracts. This primer extension activity acts preferentially on dG-rich oligodeoxynucleotides, is sensitive to RNase A pretreatment and requires all four deoxynucleotides for optimal polymerization. The extension products are also truncated by the inclusion of any one of the four dideoxynucleotides, consistent with the presence of all four bases in the S.pombe telomeric repeats. The intensity distribution of the extension products and the dideoxynucleotide termination pattern suggest that nucleotide addition is template directed, and that telomere-like sequences are added to the primers. In particular, the sequence d(CGGTTA), a variant of the S.pombe telomeric repeat, can be added directly by the in vitro activity. Partially purified S.pombe telomerase sediments as a 35S particle, suggesting that it exists in vivo as part of a large multi-protein complex. PMID- 9336463 TI - Overlapping DNA recognition motifs between Sp1 and a novel trans-acting factor within the wt1 tumour suppressor gene promoter. AB - The Wilms' tumor suppressor gene, wt1 , encodes a zinc finger transcription factor which has been shown to regulate the expression of several genes involved in cellular proliferation and differentiation. Expression of wt1 is developmentally regulated and restricted to a small set of tissues which include the fetal urogenital system, mesothelium and spleen. A highly conserved motif within the wt1 promoter, located between nucleotides -34 and -71 relative to the first transcription start site in the murine promoter, harbors consensus binding sites for Sp1 and members of the paired-box transcription factor family. Pax-2 and Pax-8 are known to enhance expression of wt1 through this conserved regulatory element. In this report, we demonstrate that Sp1 is able to bind to two sites within the 38 bp conserved region (CR). By electrophoretic mobility shift assays (EMSAs), we have identified a novel binding activity, referred to as complex D, which recognizes sequences overlapping one of the Sp1 sites in the CR. EMSA competition experiments indicate that binding of complex D and Sp1 to the CR is mutually exclusive and Sp1 is able to displace complex D binding. In situ UV crosslinking and molecular mass determinations indicate that complex D is a complex of approximately 130 kDa, consisting of at least two proteins of approximately 62 and approximately 70 kDa. Transient transfections suggest that complex D may function as an activator. PMID- 9336464 TI - Cre-mediated gene deletion in the mammary gland. AB - To delete genes specifically from mammary tissue using the Cre-lox system, we have established transgenic mice expressing Cre recombinase under control of the WAP gene promoter and the MMTV LTR. Cre activity in these mice was evaluated by three criteria. First, the tissue distribution of Cre mRNA was analyzed. Second, an adenovirus carrying a reporter gene was used to determine expression at the level of single cells. Third, tissue specificity of Cre activity was determined in a mouse strain carrying a reporter gene. In adult MMTV-Cre mice expression of the transgene was confined to striated ductal cells of the salivary gland and mammary epithelial cells in virgin and lactating mice. Expression of WAP-Cre was only detected in alveolar epithelial cells of mammary tissue during lactation. Analysis of transgenic mice carrying both the MMTV-Cre and the reporter transgenes revealed recombination in every tissue. In contrast, recombination mediated by Cre under control of the WAP gene promoter was largely restricted to the mammary gland but occasionally observed in the brain. These results show that transgenic mice with WAP-Cre but not MMTV-Cre can be used as a powerful tool to study gene function in development and tumorigenesis in the mammary gland. PMID- 9336466 TI - Functional significance of the TATA element major groove in transcription initiation by RNA polymerase II. AB - The binding of TFIID to the TATA element initiates assembly of a preinitiation complex and thus represents one of the most important steps for transcriptional regulation. The fact that the TATA binding protein (TBP), a subunit of TFIID, exclusively contacts the minor groove of the TATA element led us to ask whether the major groove of the TATA element plays any role in transcription initiation or its regulation. Our results show that modifications of the major groove of the TATA element in the adenovirus major late promoter have no effect on TFIID binding affinity or on transcription in a cell-free system reconstituted with purified factors. However, major groove modifications do decrease the levels of both basal and activator-mediated transcription in unfractionated nuclear extracts, indicating that the intact structure of the major groove of the TATA element is functionally important for transcription initiation in a more physiological context. PMID- 9336467 TI - Convergence of TNFalpha and IFNgamma signalling pathways through synergistic induction of IRF-1/ISGF-2 is mediated by a composite GAS/kappaB promoter element. AB - The molecular basis for the well known synergistic biological effects of tumor necrosis factor alpha (TNFalpha) and interferon gamma (IFNgamma) is still poorly understood. This report demonstrates that expression of interferon-regulatory factor 1 (IRF-1), also known as interferon-stimulated-gene factor 2 (ISGF-2), is synergistically induced by these cytokines. The induction is a primary transcriptional response that occurs rapidly without a requirement for new protein synthesis. Synergism is mediated by a novel composite element in the IRF 1 promoter that includes an IFNgamma-activation site (GAS) overlapped by a non consensus site for nuclear factor kappa B (NFkappaB). These sequences are bound strongly by signal transducer and activator of transcription 1 (STAT-1) and weakly by the p50/p65 heterodimer form of NFkappaB, respectively. However, the binding of STAT-1 and NFkappaB to the GAS/kappaB element in vitro seems to be mutually exclusive and independent. Synergistic induction of IRF-1 is likely to be an important early step in regulatory networks critical to the synergism of TNFalpha and IFNgamma. The GAS/kappaB element may mediate synergistic transcriptional induction of IRF-1 by other pairs of ligands that together activate NFkappaB and STAT family members. Other genes are likely to contain this motif and be regulated similarly. PMID- 9336468 TI - An ultraviolet crosslink in the hammerhead ribozyme dependent on 2-thiocytidine or 4-thiouridine substitution. AB - The hammerhead domain is one of the smallest known ribozymes. Like other ribozymes it catalyzes site-specific cleavage of a phosphodiester bond. The hammerhead ribozyme has been the subject of a vast number of biochemical and structural studies aimed at determining the structure and mechanism of cleavage. Recently crystallographic analysis has produced a structure for the hammerhead. As the hammerhead is capable of undergoing cleavage within the crystal, it would appear that the crystal structure is representative of the catalytically active solution structure. However, the crystal structure conflicts with much of the biochemical data and reveals a catalytic metal ion binding site expected to be of very low affinity. Clearly, additional studies are needed to reconcile the discrepancies and provide a clear understanding of the structure and mechanism of the hammerhead ribozyme. Here we demonstrate that a unique crosslink can be induced in the hammerhead with 2-thiocytidine or 4-thiouridine substitution at different locations within the conserved core. Generation of the same crosslink with different modifications at different positions suggests that the structure trapped by the crosslink may be relevant to the catalytically active solution structure of the hammerhead ribozyme. As this crosslink appears to be incompatible with the crystal structure, this provides yet another indication that the active solution and crystal structures may differ significantly. PMID- 9336469 TI - Mapping frequencies of endogenous oxidative damage and the kinetic response to oxidative stress in a region of rat mtDNA. AB - Genomic DNA is constantly being damaged and repaired and our genomes exist at lesion equilibrium for damage created by endogenous mutagens. Mitochondrial DNA (mtDNA) has the highest lesion equilibrium frequency recorded; presumably due to damage by H2O2 and free radicals generated during oxidative phosphorylation processes. We measured the frequencies of single strand breaks and oxidative base damage in mtDNA by ligation-mediated PCR and a quantitative Southern blot technique coupled with digestion by the enzymes endonuclease III and formamidopyrimidine DNA glycosylase. Addition of 5 mM alloxan to cultured rat cells increased the rate of oxidative base damage and, by several fold, the lesion frequency in mtDNA. After removal of this DNA damaging agent from culture, the single strand breaks and oxidative base damage frequency decreased to levels slightly below normal at 4 h and returned to normal levels at 8 h, the overshoot at 4 h being attributed to an adaptive up-regulation of mitochondrial excision repair activity. Guanine positions showed the highest endogenous lesion frequencies and were the most responsive positions to alloxan-induced oxidative stress. Although specific bases were consistently hot spots for damage, there was no evidence that removal of these lesions occurred in a strand-specific manner. The data reveal non-random oxidative damage to several nucleotides in mtDNA and an apparent adaptive, non-strand selective response for removal of such damage. These are the first studies to characterize oxidative damage and its subsequent removal at the nucleotide level in mtDNA. PMID- 9336470 TI - Nicking is asynchronous and stimulated by synapsis in 12/23 rule-regulated V(D)J cleavage. AB - The first step in DNA cleavage at V(D)J recombination signals by RAG1 and RAG2 is creation of a nick at the heptamer/coding flank border. Under proper conditions in vitro the second step, hairpin formation, requires two signals with spacers of 12 and 23 bp, a restriction referred to as the 12/23 rule. Under these conditions hairpin formation occurs at the two signals at or near the same time. In contrast, we find that under the same conditions nicking occurs at isolated signals and hence is not subject to the 12/23 rule. With two signals the nicking events are not concerted and the signal with a 12 bp spacer is usually nicked first. However, the extent and rate of nicking at a given signal are diminished by mutations of the other signal. The appearance of DNA nicked at both signals is stimulated by more than an order of magnitude by the ability of the signals to synapse, indicating that synapsis accelerates nicking and often precedes it. These observations allow formulation of a more complete model of catalysis of DNA cleavage and how the 12/23 rule is enforced. PMID- 9336471 TI - Phase imaging of moving DNA molecules and DNA molecules replicated in the atomic force microscope. AB - Phase imaging with a tapping mode atomic force microscope (AFM) has many advantages for imaging moving DNA and DNA-enzyme complexes in aqueous buffers at molecular resolution. In phase images molecules can be resolved at higher scan rates and lower forces than in height images from the AFM. Higher scan rates make it possible to image faster processes. At lower forces the molecules are imaged more gently. Moving DNA molecules are also resolved more clearly in phase images than in height images. Phase images in tapping mode AFM show the phase difference between oscillation of the piezoelectric crystal that drives the cantilever and oscillation of the cantilever as it interacts with the sample surface. Phase images presented here show moving DNA molecules that have been replicated with Sequenase in the AFM and DNA molecules tethered in complexes with Escherichia coli RNA polymerase. PMID- 9336472 TI - Escherichia coli OxyR modulation of bacteriophage Mu mom expression in dam+ cells can be attributed to its ability to bind hemimethylated Pmom promoter DNA. AB - Transcription of the bacteriophage Mu mom operon is strongly repressed by the host OxyR protein in dam - but not dam + cells. In this work we show that the extent of mom modification is sensitive to the relative levels of the Dam and OxyR proteins and OxyR appears to modulate the level of mom expression even in dam + cells. In vitro studies demonstrated that OxyR is capable of binding hemimethylated P mom , although its affinity is reduced slightly compared with unmethylated DNA. Thus, OxyR modulation of mom expression in dam + cells can be attributed to its ability to bind hemimethylated P mom DNA, the product of DNA replication. PMID- 9336473 TI - The contribution of thymine-thymine interactions to the stability of folded dimeric quadruplexes. AB - The loop of four thymines in the sodium form of the dimeric folded quadruplex [d(G3T4G3)]2 assumes a well-defined structure in which hydrogen bonding between the thymine bases appears to contribute to the stability and final conformation of the quadruplex. We have investigated the importance of the loop interactions by systematically replacing each thymine in the loop with a cytosine. The quadruplexes formed by d(G3CT3G3), d(G3TCT2G3), d(G3T2CTG3) and d(G3T3CG3) in the presence of 150 mM Na+ were studied by gel mobility, circular dichroism and 1H NMR spectroscopy. The major species formed by d(G3CT3G3), d(G3TCT2G3) and d(G3T3CG3) at 1 mM strand concentration at neutral pH is a dimeric folded quadruplex. d(G3T2CTG3) has anomalous behaviour and associates into a greater percentage of linear four-stranded quadruplex than the other three oligonucleotides at neutral pH and at the same concentration. The linear four stranded quadruplex has a greater tendency to oligomerize to larger ill-defined structures, as demonstrated by broad 1H NMR resonances. At pH 4, when the cytosine is protonated, there is a greater tendency for each of the oligonucleotides to form some four-stranded linear quadruplex, except for d(G3T2CTG3), which has the reverse tendency. The experimental results are discussed in terms of hydrogen bonding within the thymine loop. PMID- 9336474 TI - Interaction of the phage T4 Dam DNA-[N6-adenine] methyltransferase with oligonucleotides containing native or modified (defective) recognition sites. AB - The DNA-[N 6-adenine]-methyltransferase (Dam MTase) of phage T4 catalyzes methyl group transfer from S-adenosyl-l-methionine (AdoMet) to the N6-position of adenine in the palindromic sequence, GATC. We have used a gel shift assay to monitor complex formation between T4 Dam and various synthetic duplex oligonucleotides, either native or modified/defective. The results are summarized as follows. (i) T4 Dam bound with approximately 100-fold higher affinity to a 20mer specific (GATC-containing) duplex containing the canonical palindromic methylation sequence, GATC, than to a non-specific duplex containing another palindrome, GTAC. (ii) Compared with the unmethylated duplex, the hemimethylated 20mer specific duplex had a slightly increased ( approximately 2-fold) ability to form complexes with T4 Dam. (iii) No stable complex was formed with a synthetic 12mer specific (GATC-containing) duplex, although T4 Dam can methylate it. This indicates that there is no relation between formation of a catalytically competent 12mer-Dam complex and one stable to gel electrophoresis. (iv) Formation of a stable complex did not require that both strands be contiguous or completely complementary. Absence of a single internucleotide phosphate strongly reduced complex formation only when missing between the T and C residues. This suggests that if T4 Dam makes critical contact(s) with a backbone phosphate(s), then the one between T and C is the only likely candidate. Having only one half of the recognition site intact on one strand was sufficient for stable complex formation provided that the 5'G.C base-pairs be present at both ends of the palindromic, GATC. Since absence of either a G or C abolished T4 Dam binding, we conclude that both strands are recognized by T4 Dam. PMID- 9336475 TI - Position effects in mice carrying a lacZ transgene in cis with the beta-globin LCR can be explained by a graded model. AB - We studied transgenic mice carrying the lacZ reporter gene linked to the erythroid-specific beta-globin promoter and beta-globin locus control region (LCR). Previously, we had demonstrated that the total level of expression of beta galactosidase enzyme, which is the product of the lacZ gene, varies widely between different transgenic mice due to position effects at the sites of transgene integration. Here, using the X-gal based in situ assay for beta galactosidase activity, we found that the percent erythroid cells that expressed the transgene also varied widely between the mice. Moreover, a kinetic analysis showed that the average beta-galactosidase content per expressing cell varied both between samples of different transgenic descent and between erythroid cells within each sample, demonstrating that the variable expression of this lacZ transgene was being controlled in a graded manner. These results suggest that the beta-globin LCR enhancers function through a graded model, which is described, rather than the binary mechanism that has been proposed previously for other enhancers. PMID- 9336476 TI - Sequence walkers: a graphical method to display how binding proteins interact with DNA or RNA sequences. AB - A graphical method is presented for displaying how binding proteins and other macromolecules interact with individual bases of nucleotide sequences. Characters representing the sequence are either oriented normally and placed above a line indicating favorable contact, or upside-down and placed below the line indicating unfavorable contact. The positive or negative height of each letter shows the contribution of that base to the average sequence conservation of the binding site, as represented by a sequence logo. These sequence 'walkers' can be stepped along raw sequence data to visually search for binding sites. Many walkers, for the same or different proteins, can be simultaneously placed next to a sequence to create a quantitative map of a complex genetic region. One can alter the sequence to quantitatively engineer binding sites. Database anomalies can be visualized by placing a walker at the recorded positions of a binding molecule and by comparing this to locations found by scanning the nearby sequences. The sequence can also be altered to predict whether a change is a polymorphism or a mutation for the recognizer being modeled. PMID- 9336477 TI - Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus. AB - The introduction of high molecular weight DNA into mammalian cells is useful for gene expression studies. However, current transfection strategies are inefficient, necessitating propagation of stable DNA transformants prior to analysis of gene expression. Here we demonstrate that transient lipid-mediated DNA transfection can be used to assess gene expression from yeast artificial chromosomes (YACs) containing the 230 kb cystic fibrosis transmembrane conductance regulator gene ( CFTR ) and Escherichia coli lacZ . We also show that psoralen-UV inactivated adenovirus significantly enhances transfection efficiency. The ability to deliver high molecular weight DNA using lipid-mediated transfection should expedite the analysis of large human genes contained within artificial chromosome vectors. PMID- 9336478 TI - A rapid RT-PCR based method to isolate complementary DNA fragments flanking retrovirus integration sites. AB - Proto-oncogenes in retrovirally induced myeloid mouse leukemias are frequently activated following retroviral insertion. The identification of common virus integration sites (VISs) and isolation of the transforming oncogene is laborious and time consuming. We established a rapid and simple PCR based procedure which facilitates the identification of VISs and novel proto-oncogenes. Complementary DNA fragments adjacent to retrovirus integration sites were selectively isolated by applying a reverse transcriptase (RT) reaction using an oligo(dT)-adaptor primer, followed by PCR using the adaptor sequence and a retrovirus long terminal repeat (LTR) specific primer. Multiple chimeric cDNA fragments suitable for Southern and northern blot analysis were isolated. PMID- 9336479 TI - Detection and measurement of PCR bias in quantitative methylation analysis of bisulphite-treated DNA. AB - Methylation analysis of individual cytosines in genomic DNA can be determined quantitatively by bisulphite treatment and PCR amplification of the target DNA sequence, followed by restriction enzyme digestion or sequencing. Methylated and unmethylated molecules, however, have different sequences after bisulphite conversion. For some sequences this can result in bias during the PCR amplification leading to an inaccurate estimate of methylation. PCR bias is sequence dependent and often strand-specific. This study presents a simple method for detection and measurement of PCR bias for any set of primers, and investigates parameters for overcoming PCR bias. PMID- 9336481 TI - SPECIAL SECTION: Perspectives of the Scientific Community on the Status of Ecological Risk Assessment AB - / Views from a wide variety of practicing environmental professionals on the current status of ecological risk assessment (ERA) indicate consensus and divergence of opinion on the utility and practice of risk assessment. Central to the debate were the issues of whether ERA appropriately incorporates ecological and scientific principle into its conceptual paradigm. Advocates argue that ERA effectively does both, noting that much of the fault detractors find with the process has more to do with its practice than its purpose. Critics argue that failure to validate ERA predictions and the tendency to over-simplify ecological principles compromise the integrity of ERA and may lead to misleading advice on the appropriate responses to environmental problems. All authors felt that many improvements could be made, including validation, better definition of the ecological questions and boundaries of ERA, improved harmonization of selected methods, and improvements in the knowledge base. Despite identified deficiencies, most authors felt that ERA was a useful process undergoing evolutionary changes that will inevitably determine the range of environmental problems to which it can be appropriately applied. The views expressed give ERA a cautious vote of approval and highlight many of the critical strengths and weaknesses in one of our most important environmental assessment tools.KEY WORDS: Ecological risk assessment; Ecology; Probability PMID- 9336482 TI - PROFILE: Multiattribute Utility Analysis as a Framework for Public Participation in Siting a Hazardous Waste Management Facility AB - / In an attempt to facilitate the resolution of contentious environmental problems, public and private organizations are experimenting with collaborative approaches wherein stakeholders participate in the decision-making process. A dilemma for the design of collaborative approaches is the technical complexity of many environmental problems. How can members of the public play a meaningful role in decisions that involve complicated scientific arguments?This paper describes a public participation exercise in which stakeholders used an approach based on multiattribute utility analysis to select a site for a hazardous waste management facility. The key to success was the ability to separate and address two types of judgments inherent in environmental decisions-technical judgments regarding the likely consequences of alternative choices and value judgments regarding the importance or seriousness of those consequences. The approach enabled technical specialists to communicate the essential technical considerations and allowed stakeholders to establish the value judgments for the decision. Although rarely used in public participation, the multiattribute utility approach appears to provide a useful framework for the collaborative resolution of complex environmental decision problems.KEY WORDS: Multiattribute utility analysis; Public involvement; Collaboration; Dispute resolution; Environmental management PMID- 9336480 TI - A method for generating subtractive cDNA libraries retaining clones containing repetitive elements. AB - Here we describe a two-stepped photobiotin-based procedure to enrich a target (canine retinal) cDNA library for tissue specific clones without removing those containing repetitive ( SINE ) elements, despite the presence of these elements in the driver population. In a first hybridization excess SINE elements were hybridized to a driver (canine cerebellar) cDNA. In a second hybridization target cDNA was added to this reaction. The resulting cDNA library was enriched for retinal specific clones, but contained the same ratio of clones with SINE elements found in the unsubtracted library. PMID- 9336483 TI - Water Resource Protection in Taiwan: An Evaluation of the Taipei Water Management Commission AB - / This study evaluates the institutional capacity and performance of the Taipei Water Management Commission. The commission, which manages the Taipei Water Special Area-one of 95 such areas in Taiwan and the only one managed by a supervisory agency-has established a record of water conservation that suggests its utility as a model for managing other protected water resources areas in Taiwan. However, its present institutional structure limits its ability deliver on its mandate. The study identifies a number of problems related to the commission's current institutional structure that need to be addressed if the commission is to serve as a viable model for managing other protected water resource areas in Taiwan.KEY WORDS: Water resources management; Commissions; Institutional capacity; Taiwan PMID- 9336484 TI - Paradise Threatened: Land Use and Erosion on St. John, US Virgin Islands AB - / Rapid development and the concomitant increases in erosion and sedimentation are believed to threaten the reefs and other marine resources that are a primary attraction of St. John and Virgin Islands National Park. Average annual sediment yields from undeveloped areas were estimated from a sediment pond and a mangrove swamp as less than 20 and less than 40 t/km2/yr, respectively. Geomorphic evidence indicates that plantation agriculture during the 18th and 19th centuries did not cause severe erosion. Since about 1950 there has been rapid growth in roads and development due to increasing tourism and second-home development. Our field investigations identified the approximately 50 km of unpaved roads as the primary source of anthropogenic sediment. Field measurements of the road network in two catchments led to the development of a vector-based GIS model to predict road surface erosion and sediment delivery. We estimate that road erosion has caused at least a fourfold increase in island-wide sediment yields and that current sedimentation rates are unprecedented. Paving the dirt roads and implementing standard sediment control practices can greatly reduce current sediment yields and possible adverse effects on the marine ecosystems surrounding St. John.KEY WORDS: Erosion; Sediment yield; Roads; Dry tropics; Development PMID- 9336485 TI - Environmental Reporting by the Fortune 50 Firms AB - / The extent and use of industry-reported environmental data are increasing, warranting an in-depth analysis of this information. This paper reviews the environmental reporting guidelines issued by several business and nonprofit organizations and evaluates the environmental reports published by the Fortune 50 companies, half of which publish reports. After describing the history of environmental reporting and the content of the guidelines, a comparative evaluation is made to indicate the types of companies producing reports, the topics reported, the intended audiences, the scope and depth of the material reported, and the effectiveness of the reports as communication devices. These reports are mechanisms to enhance a firm's image, public relations, and marketing and are aimed largely at concerned individuals, affected communities, and investors. Significant differences in the content and the depth of reports are seen as firms report on topics that are perceived by the public as high risks. The most complete reports are published by industries with poor or controversial public images, e.g., the chemical and timber industries. Still, no report provided information that was sufficient for comprehensive or comparative analyses of environmental performance. Recommendations are provided to increase the quality and effectiveness of environmental reporting.KEY WORDS: Communication; Environmental management; Performance reporting; Reporting; Stakeholder PMID- 9336486 TI - RESEARCH: Theory in Practice: Applying Participatory Democracy Theory to Public Land Planning AB - / Application of participatory democracy theory to public participation in public land planning, while widely advocated, has not been closely examined. A case study is used here to explicate the application of participatory democracy concepts to public participation in public land planning and decision making. In this case, a Bureau of Land Management resource area manager decided to make a significant shift from the traditional public involvement process to a more participatory method-coordinated resource management (CRM). This case was assessed using document analysis, direct observation of CRM meetings, questionnaires, and interviews of key participants. These sources were used to examine the CRM case using participatory democracy concepts of efficacy, access and representation, continuous participation throughout planning, information exchange and learning, and decision-making authority. The case study suggests that social deliberation in itself does not ensure successful collaboration and that establishing rules of operation and decision making within the group is critical. Furthermore, conflicts between the concept of shared decision-making authority and the public land management agencies' accountability to Congress, the President, and the courts need further consideration.KEY WORDS: Case study; Coordinated resource management; Public participation; Administrative discretion; Representation; Consensus; Collaboration PMID- 9336487 TI - Landscape Structure and Historical Processes Along Diked European Valleys: A Case Study of the Arc/Isere Confluence (Savoie, France) AB - / The valleys of European piedmonts constitute changing narrow corridors within which water, matter, nutrients, energy, and species flow. The dispersion patterns of these flows have been significantly disturbed since the end of the 18th century. Thus, western European valleys have been changed into complex mosaics by implementation of socioeconomical programs. In order to define future actions allowing the preservation of this "ecocomplex" (Blandin and Lamotte 1988), it is necessary to gather precise information of the landscape dynamics. Hence, the study of the European river valleys must be based upon two major steps: (1)the analysis of the present landscape with suitable remote sensing techniques, allowing us to map the complex mosaic of narrow corridors; and (2) the analysis of temporal landscape development patterns since the first engineering works that have transformed the braided channel system. In this paper, the efficiency of the addition of two techniques is highlighted: (1) the "wavelet merging method" from multispectral and panchromatic SPOT images for the floodplain land-cover mapping, and (2) the historical reconstruction techniques from old maps and archive documents in order to analyze the cumulative impacts of engineering works on landscape diversity. To illustrate the method, a particularly complex case study is chosen: the Arc/Isere confluence (downstream from Albertville, Savoie, France). Remote sensing, field survey, and historical reconstruction allowed us to distinguish two types of spatial units: (1) the "functional sets" characterized by independent state factors (edaphic, hydrological, and topographic) and supporting a limited number of vegetation types (spontaneous or cultivated), and (2) the functional unit (= ecotope), which corresponds to a unique combination of vegetation type/functional set.KEY WORDS: Landscape ecology; Floodplains; Vegetation; Land use; Remote sensing; Historical reconstruction; Mapping; Merging methods; Human impacts; Alps; Isere river; France PMID- 9336488 TI - Long Profile Evolution of a Mountain Stream in Relation to Gravel Load Management: Example of the Middle Giffre River (French Alps) AB - / Since the 1970s, many French riverbeds have been incised by more than 1 m. This generalized phenomenon, apparently irreversible, is rightly considered as alarming. However, our study of the Giffre, a sixth-order high-energy river draining an intramountain plain in the northern French Alps, leads us to qualify this general opinion. Although the Giffre underwent considerable incision as early as the 1960s (-1.16 m between 1912 and 1988, for a total sediment loss of 2 x 10(6) m3) following extensive gravel extraction from the channel, this evolution appears to be reversed today, showing that this river is capable of rehabilitating itself. The watershed supplies the river with 50,000 m3/yr of material and part of this load (30,000 m3/yr) is extracted. Although it is theoretically possible to reverse this phenomenon, it is unacceptable for the local economy as man-made installations unadapted to flooding were developed along the river during the period of incision. Today, the development policy is in conflict with the maintenance and the preservation of natural sediment transport and deposition.KEY WORDS: Bed incision; Gravel load management; Giffre River; France PMID- 9336489 TI - Morphology of a Wetland Stream AB - / Little attention has been paid to wetland stream morphology in the geomorphological and environmental literature, and in the recently expanding wetland reconstruction field, stream design has been based primarily on stream morphologies typical of nonwetland alluvial environments. Field investigation of a wetland reach of Roaring Brook, Stafford, Connecticut, USA, revealed several significant differences between the morphology of this stream and the typical morphology of nonwetland alluvial streams. Six morphological features of the study reach were examined: bankfull flow, meanders, pools and riffles, thalweg location, straight reaches, and cross-sectional shape. It was found that bankfull flow definitions originating from streams in nonwetland environments did not apply. Unusual features observed in the wetland reach include tight bends and a large axial wavelength to width ratio. A lengthy straight reach exists that exceeds what is typically found in nonwetland alluvial streams. The lack of convex bank point bars in the bends, a greater channel width at riffle locations, an unusual thalweg location, and small form ratios (a deep and narrow channel) were also differences identified. Further study is needed on wetland streams of various regions to determine if differences in morphology between alluvial and wetland environments can be applied in order to improve future designs of wetland channels.KEY WORDS: Stream morphology; Wetland restoration; Wetland creation; Bankfull; Pools and riffles; Meanders; Thalweg PMID- 9336490 TI - ENVIRONMENTAL AUDITING: Demonstration of Line Transect Methodologies to Estimate Urban Gray Squirrel Density AB - / Because studies estimating density of gray squirrels (Sciurus carolinensis) have been labor intensive and costly, I demonstrate the use of line transect surveys to estimate gray squirrel density and determine the costs of conducting surveys to achieve precise estimates. Density estimates are based on four transects that were surveyed five times from 30 June to 9 July 1994. Using the program DISTANCE, I estimated there were 4.7 (95% CI = 1.86-11.92) gray squirrels/ha on the Clemson University campus. Eleven additional surveys would have decreased the percent coefficient of variation from 30% to 20% and would have cost approximately $114. Estimating urban gray squirrel density using line transect surveys is cost effective and can provide unbiased estimates of density, provided that none of the assumptions of distance sampling theory are violated.KEY WORDS: Bias; Density; Distance sampling; Gray squirrel; Line transect; Sciurus carolinensis. PMID- 9336492 TI - Clinical evaluation and management of aneurysmal subarachnoid hemorrhage. AB - As neuroradiologic techniques become more critical to the care of patients suffering from aneurysmal subarachnoid hemorrhage, a thorough understanding of the natural history and medical management of this disorder by neuroradiologists is required to insure appropriate diagnosis and therapy. This article addresses the medical and perioperative management of subarachnoid hemorrhage, with an emphasis on features relevant to neuroradiologic diagnosis and treatment. PMID- 9336491 TI - Epidemiology of aneurysmal subarachnoid hemorrhage. AB - An estimated 1% to 5% of adults have a cerebral aneurysm. Each year, approximately 1 in 10,000 North Americans suffer an aneurysmal subarachnoid hemorrhage, with greater than 50% combined morbidity and mortality. Cerebral aneurysm formation and rupture is associated with a variety of factors, including increasing age, female gender, hypertension, alcohol, smoking, and genetic factors. PMID- 9336494 TI - The role of CT following aneurysmal rupture. AB - Computed tomography is a highly effective method of detecting subarachnoid blood if performed early after aneurysmal rupture, being 95% to 98% positive when lumbar puncture is positive. The localization of the blood defines the location of the aneurysm in approximately 80% of cases. Contrast enhanced computed tomography demonstrates the aneurysm in 75% of cases with the aneurysm is greater than 5 millimeters in size. Computed tomography angiography defines the aneurysm in up to 96% of cases. The amount of subarachnoid blood correlates with the development of vasospasm; cerebral perfusion can be further evaluated with xenon enhanced computed tomography. PMID- 9336493 TI - Neuro-ophthalmologic aspects of aneurysms. AB - The visual pathways and the ocular motor cranial nerves are frequently injured by expanding cerebral aneurysms. Neuro-ophthalmologic signs and symptoms may be the only indications of an aneurysm prior to rupture. Acute or chronic visual loss may herald an aneurysm prior to rupture. Acute or chronic visual loss may herald an aneurysm in the carotidophthalmic, supra clinoid carotid, internal carotid bifurcation, or anterior communicating artery distributions. Diplopia and retro orbital pain may be warning signs that precede the discovery of a posterior communicating, basilar, or cavernous sinus aneurysm. PMID- 9336495 TI - Magnetic resonance imaging of intracranial aneurysms. AB - Intracranial aneurysms are lesions with high morbidity and mortality, but in most cases represent treatable disease. Magnetic resonance (MR) imaging and in some cases MR angiography can make valuable contributions to their diagnosis and characterization. The ultimate tool for imaging these lesions, however, remains catheter angiography. This article focuses on saccular aneurysms, with a brief discussion on atherosclerotic fusiform aneurysms and mycotic aneurysms. PMID- 9336496 TI - Angiographic evaluation of aneurysms affecting the central nervous system. AB - This article discusses the indications for, techniques of, and risks associated with cerebral angiography. Various types of aneurysms are described, and there is a discussion of the angiographic features of aneurysms in specific locations. Subarachnoid hemorrhage, post-subarachnoid hemorrhage vasospasm, and the role of post-operative angiography are also covered. PMID- 9336497 TI - Neuroendovascular management of intracranial aneurysms. AB - Intracranial aneurysms are a common disease constituting a significant health problem worldwide. Aneurysmal subarachnoid hemorrhage, cerebral aneurysms, classification of aneurysms, and management of aneurysms are discussed. PMID- 9336498 TI - Surgical treatment of intracranial aneurysms. AB - Intracranial aneurysms are dangerous lesions which may produce sudden death or neurological devastation in people who are often otherwise completely healthy. The definitive treatment for most of these lesions in microsurgical repair, and the cure rate is quite high. The indications for surgery include an accessible aneurysm in a patient who is healthy enough to tolerate general anesthesia and a major intracranial procedure. The timing of surgery following subarachnoid hemorrhage is still controversial, but early surgery is currently recommended for patients who are alert. Preoperative imaging studies are crucial in defining the precise anatomy of the aneurysm and any associated lesions and anomalies. Surgical approaches and techniques, anesthetic considerations, postoperative radiographic evaluation, overall results, and complications are discussed in this article. PMID- 9336499 TI - Extradural aneurysms. AB - Extradural aneurysms have distinct characteristics from their intradural counterparts. Most extradural aneurysms cannot be treated by direct surgical exposure and clip ligation or by direct endovascular means without parent vessel sacrifice. Arterial occlusion with or without bypass grafting remains the traditional treatment. Controversy about the "best" or "proper" technique of arterial balloon test occlusion is rivaled only by that of the necessity for bypass grafting when apparent tolerance for arterial occlusion has been demonstrated. PMID- 9336500 TI - Unusual aneurysms. AB - Cerebral aneurysms can present in a variety of special circumstances. Aneurysmal subarachnoid hemorrhage (SAH) can complicate systemic or neoplastic disease, head injuries, arterial dissection, and other cerebrovascular conditions. Aneurysms associated with non-saccular configuration or giant size can make surgical or endovascular intervention difficult if not impossible. This article will review these uncommon aneurysms and their management. PMID- 9336502 TI - Terrorism in America. An evolving threat. AB - The response of the United States to a perceived terrorist threat is dichotomous. The hyperbole and exaggeration often displayed by the media and general public lies in stark contrast to the relative indifference with which terrorism is regarded by the medical community. Quantitating the true nature of the terrorist threat in the United States is difficult, as it is not only poorly defined but rapidly changing. The intent of this commentary is to define what constitutes terrorism and what specific threats exist, including conventional, nuclear, biological, and chemical weapons. We will then outline recommendations for modest changes in our disaster medical planning to better prepare for these threats. Special attention will be directed at the emergence of nonconventional weapon use by terrorist organizations and how this might affect the civilian medical community. PMID- 9336501 TI - Neuroradiologic diagnosis and treatment of vasospasm. AB - For survivors of aneurysmal subarachnoid hemorrhage, cerebral vasospasm significantly contributes to its morbidity and mortality by causing delayed ischemic neurological deficit. Noninvasive evaluation with computed tomography, transcranial doppler and single photon emission computerized tomography helps guide clinical decisions. Endovascular therapy of symptomatic vasospasm with balloon angioplasty and to a lesser extent with intraarterial papaverine infusion has emerged as an important treatment adjunct to neurosurgical medical and operative management. Early and aggressive treatment with balloon angioplasty has resulted in sustained clinical improvement in about two-thirds of patients suffering from neurological deficits attributable to vasospasm. Encouraging long term clinical and transcranial artery damage following angioplasty. Despite balloon angioplasty's 2% to 5% peri-procedure mortality rate, it remains under used. PMID- 9336503 TI - More guns and younger assailants. A combined police and trauma center study. AB - OBJECTIVE: To test the hypothesis that guns have become the weapon of choice for assaults and that both assailants and victims have become progressively younger. DESIGN: Retrospective review of trauma center and police department data sources. SETTING: Regional trauma center with university affiliation; municipal police department. SUBJECTS: Victims of assault with a deadly weapon from 3 discontiguous years. MAIN OUTCOME MEASURES: Age of assailant and victim, type of injury, frequency of blunt vs penetrating injury. RESULTS: From June 1991 to May 1992 and June 1993 to May 1994, the incidence of penetrating trauma increased from 27% to 35% of trauma center admissions (chi 2 test; P < .001). During the period from June 1985 to May 1994, assault with a deadly weapon increased by 220% and firearms became the most common assault mechanism (from 32% to 54%; chi 2 test; P < .001). Assailants using guns became significantly younger, with the percentage of assailants aged 11 to 20 years increasing from 24% to 47% (chi 2 test; P = .001). The ages of assault victims also decreased (P < .003), but were more evenly distributed across age categories. CONCLUSIONS: The incidence of penetrating trauma has increased in both absolute numbers and in relative proportion to blunt trauma. Firearms have become the weapon of choice and the single largest group of assailants are 11 to 20 years of age. The use of concurrent police and trauma center databases provides a more cogent basis for developing effective violence prevention strategies. PMID- 9336504 TI - Portal vein gas, a changing clinical entity. Report of 7 patients and review of the literature. AB - OBJECTIVE: To assess the clinical significance of portal vein gas (PVG) demonstrated by computed tomography (CT). DESIGN: Review of medical records. SETTING: Three network-affiliated hospitals providing both primary community based and tertiary services. METHODS: Review of diagnosis, clinical circumstances, and significance of PVG in 7 patients detected by CT during a 3 year period in 3 affiliated hospitals. RESULTS: Four of 7 patients underwent laparotomy; 1 patient refused surgery. Two patients were treated with intravenous antibiotics only and had uneventful clinical courses. Of the 3 patients who died, 1 refused and 2 underwent laparotomy. CONCLUSIONS: This series indicates that more sensitive imaging and more widespread use of endoscopic retrograde cholangiopancreatography, colonoscopy, and liver transplantation have changed the clinical presentation of PVG; PVG may be found in various clinical settings that do not mandate laparotomy; and the significance of PVG must be derived from the clinical context of the individual patient. PMID- 9336505 TI - Combined liver-total bowel transplantation has no immunologic advantage over total bowel transplantation alone. A prospective study in a porcine model. AB - BACKGROUND: Rejection remains a major obstacle to successful bowel transplantation in humans. It has been suggested that a simultaneous liver transplant would shield the bowel graft from immunologic attack, but the liver shortage would be aggravated. In a preclinical model, we studied the influence of simultaneous liver grafting by comparing the incidence of early bowel rejection after combined liver-total (small- and large-) bowel transplants vs total bowel transplants alone. METHODS: We assessed the incidence of early post-transplant rejection, graft-vs-host disease, and infection after combined liver-total bowel transplants (group 1, n = 10) and total bowel transplants alone (group 2, n = 9) in outbred Yorkshire Landrace pigs. Liver and bowel grafts were transplanted orthotopically with portal vein drainage after recipient hepatectomy (group 1) and total enterectomy (groups 1 and 2). Posttransplant immunosuppression was performed with intravenous tacrolimus (whole blood levels, 15 to 30 ng/mL) and prednisolone. In groups 1 and 2, bowel biopsy specimens from the ileostomy were obtained daily. In group 1, liver biopsy specimens were obtained weekly. Rejection was graded according to a 4-point scoring system (none, mild, moderate, and severe). RESULTS: Overall graft survival at days 7, 14, and 21 was 89%, 44%, and 11%, respectively, in group 1 vs 100%, 100%, and 86%, respectively, in group 2 (P < .001). Death rates owing to (irreversible) rejection at days 7, 14, and 21 were 0% in groups 1 and 2 (P = .48). Grading of bowel rejection episodes, based on the results of daily biopsy specimens, was not significantly different between the groups whether on individual days or overall. In group 1, the incidence of liver rejection episodes was as high as 66% (day 14 and at autopsy). At autopsy, generalized graft-vs-host disease (skin, native intestine, and native liver) was noted in 55% of group 1 and 43% of group 2 pigs (P = .55). Graft-vs-host disease was noted concurrently with rejection episodes of the liver or bowel grafts. CONCLUSIONS: Simultaneous liver grafting did not further reduce the incidence of early bowel rejection or graft-vs-host disease when compared with total bowel transplants alone. Based on the results of this preclinical study, simultaneous liver grafting is not indicated for patients with short-bowel syndrome and normal liver function. PMID- 9336506 TI - Treatment of pneumonia in mechanically ventilated trauma patients. Results of a prospective trial. AB - OBJECTIVES: To determine the efficacy and magnitude of associated adverse effects of 2 different antibiotic regimens for the treatment of pneumonia in intubated surgical patients and to assay and compare blood samples and bronchoalveolar lavage fluid with respect to some host-defense parameters, especially in patients with unilateral pneumonia. DESIGN: Randomized, prospective, unblinded clinical comparison of 2 treatment arms with respect to intent to treat and clinical and microbiologically evaluable patients. SETTING: Six university surgical services in teaching hospitals with modern and well-staffed intensive care units. INTERVENTIONS: The consistency and objectively of the diagnosis of pneumonia was improved by the use of a grid of diagnostic parameters. Aggressive mechanical approaches to pneumonia in intubated surgical patients were supplemented by therapeutic use of aztreonam and vancomycin hydrochloride or combined imipenem and cilastatin sodium. RESULTS: Patients randomized to the aztreonam-vancomycin group were somewhat more ill, fared slightly better, and had fewer serious drug related side effects than did those treated with imipenem-cilastatin (all P > .05). Immunologic parameters assessed by evaluation of bronchoalveolar lavage fluid showed differences between infected pulmonary lobes and noninfected ones; some changes were also noted in patients who recovered compared with those whose pneumonia persisted or recurred. CONCLUSIONS: Clinical studies of pneumonia in surgical patients need to be stratified to assure comparability, to identify patients in whom treatment is likely to fail, and to display differences between more and less effective therapies. Studies of blood and bronchoalveolar lavage samples showed that certain local and systemic immunologic parameters correlate with clinical status and outcome. PMID- 9336507 TI - Management of bowel obstruction in patients with abdominal cancer. AB - OBJECTIVE: To determine the value of operation in patients with bowel obstruction caused by recurrent abdominal cancer. DESIGN: Retrospective case review. SETTING: The University of Connecticut Health Center, Farmington. PATIENTS: Ninety-eight patients admitted with a diagnosis of bowel obstruction and malignant neoplasm between November 1, 1987, and June 30, 1995. RESULTS: Data for 75 patients who developed a bowel obstruction within 5 years of a malignant diagnosis were analyzed. Forty-six patients (61%) were treated operatively and 29 (39%) were treated nonoperatively. The operative group included 32 patients (70%) whose obstruction was caused by carcinomatosis; 6 (19%) of these 32 patients had had at least 1 episode of previous obstruction requiring hospitalization. They had a 22% in-hospital mortality, stayed an average of 21 days in the hospital, and survived 7 +/- 6 months (mean +/- SD) after discharge; 5 (16%) had at least 1 episode of postoperative obstruction that required hospitalization. After discharge from the hospital, 53% had an excellent or good quality of life (assessed retrospectively). Of the 29 patients in the nonoperative group, 16 (55%) had carcinomatosis. These 16 patients had a 38% in-hospital mortality (6 of 16), stayed an average of 10 days in the hospital, and survived a mean of 13 +/- 9 months; 3 (19%) had at least 1 episode of recurrent obstruction requiring hospitalization. After discharge from the hospital, 6 (37%) had an excellent or good quality of life. CONCLUSION: The value of operative intervention for bowel obstruction in patients with cancer is derived from the possibility of a benign cause, not alleviation of the consequences of carcinomatosis. PMID- 9336509 TI - Total vascular exclusion of the liver during hepatic surgery. Selective use, extensive use, or abuse? AB - OBJECTIVES: To review our experience with total vascular exclusion of the liver and to assess its role in hepatic resections. DESIGN: Retrospective survey. SETTING: University hospital, a tertiary referring center for surgical liver diseases. PATIENTS: A total of 722 patients who underwent liver resections from November 1, 1981, to March 31, 1996, of whom 19 (2.6%) required total vascular exclusion because of hepatic lesions closely adherent to or infiltrating the retrohepatic vena cava or centrally located in the liver, strictly in contact with the hepatic vein convergence. MAIN OUTCOME MEASURE: chi 2 Test for qualitative data and Student t test for categorical data. RESULTS: Of the 19 resections carried out under total vascular exclusion, 6 had tumoral infiltration of the retrohepatic vena cava: in 4 cases the venous wall was partially resected, while in the remaining 2 it was completely removed and replaced with a prosthetic graft. There were no operative deaths. Of the 722 resections, 227 were major hepatectomies: 74 (32.6%) were performed after ligation of the glissonian elements for the hemiliver to be removed, without clamping of the hepatic pedicle, and a further 36 (15.8%) were performed without any preliminary vascular control. A significant reduction in intraoperative blood transfusions was achieved despite the performance of more extended operations, regardless of the technique used. CONCLUSIONS: Total vascular exclusion is a useful tool in controlling blood inflow to the liver, but true need for it during liver resection is limited. Its performance requires a well-trained team familiar with problems regarding surgical access to the inferior vena cava and prolonged occlusion of the hepatic pedicle and the inferior vena cava. PMID- 9336508 TI - The impact of surgical complications after liver transplantation on resource utilization. AB - OBJECTIVE: To evaluate the impact of surgical complications on length of stay and hospital charges after liver transplantation. DESIGN: A retrospective economic evaluation of the outcomes during initial hospitalization after liver transplantation. SETTING: University hospital treating referred patients. PATIENTS: The study population was 109 patients undergoing 111 liver transplantations during fiscal year 1993. MAIN OUTCOME MEASURES: Hospital charges and length of stay during the initial hospitalization after liver transplantation. Multivariate regression methods were used to analyze the impact of surgical complications on costs. RESULTS: Of the 111 transplantations, 30 (27%) had a surgical complication that required a return to the operating room during the initial hospitalization. The effect of a surgical complication was to increase the mean hospital charges (excluding physician charges) from $150,092 to $347,728 (difference of mean, $197,636; confidence interval of difference, $114,153 to $319,326). The median length of stay was 16 days for patients without complications and 45 days for those with complications. Univariate and multivariate models suggested that surgical complications had the greatest effect on length of stay and hospital charges among the factors studied. Complications tended to occur more frequently among patients with United Network for Organ Sharing (UNOS) status 1 (42% vs 22%), but this did not reach statistical significance (P = .09). CONCLUSIONS: Surgical complications after liver transplantation have a marked impact on the cost of the procedure. The magnitude of this effect is greater than that of UNOS status, presence of rejection, or other demographic or clinical factors studied. Complications tend to occur in the most ill patients. Identifying strategies to reduce the risk of complications, particularly in patients with UNOS status 1, likely can reduce the cost of transplantation. PMID- 9336510 TI - High-volume vs standard fluid therapy in a septic pig model. Impact on pulmonary function. AB - OBJECTIVE: To compare pulmonary function and peripheral organ blood flow in septic pigs receiving high-volume fluid resuscitation or standard-volume fluid resuscitation with similar goals in oxygen delivery. DESIGN: A prospective study comparing 2 groups of septic pigs. SETTING: A university animal research laboratory. SUBJECTS: Eleven septic pigs. INTERVENTIONS: Basal oxygen delivery was increased from 450 to 550 mL/min to at least 600 mL/min by the sixth hour and maintained for 24 hours. From a baseline pulmonary artery occlusion pressure (PAOP) measurement of approximately 6 mm Hg, the high-volume group (n = 5) was treated until a PAOP measurement of 12 mm Hg was reached and the standard-volume group (n = 6) was treated until a PAOP measurement of 8 mm Hg was reached. Blood transfusions and inotropic agents were added as necessary to reach the oxygen delivery goal. RESULTS: The high-volume group had a significantly greater positive fluid balance, greater weight gain, and a higher PAOP but similar intrapulmonary shunt and extravascular lung water as compared with the standard volume group. CONCLUSION: Resuscitation with large volumes of fluid in early sepsis with a physiological goal of a higher PAOP to augment oxygen delivery did not cause increased pulmonary edema and oxygenation deficit compared with maintenance of lower cardiac filling pressures. PMID- 9336511 TI - Is the activity of soluble CD14 enhanced following major trauma? AB - BACKGROUND: The molecule CD14 acts as a receptor for the protein-bound endotoxin (lipopolysaccharide [LPS]) complex and mediates the cellular effects of LPS. The soluble formation, sCD14, is supposed to neutralize circulating LPS (i.e., LPS antagonist) or transfer LPS effects to endothelial cells (i.e., LPS agonist). OBJECTIVE: To elucidate the release of sCD14 per se in patients with major trauma in the early posttrauma period. Our a priori hypothesis was that sCD14 release depends on the plasma LPS concentration simultaneously measured. PATIENTS: In a prospective study, 65 patients with multiple injuries (Injury Severity Score, 9 75) were enrolled. The patients were rescued by the medical helicopter service and directly admitted to our clinics. The plasma concentrations of sCD14 (enzyme immunoassay) and LPS (chromogenic limulus amebocyte lysate test) were analyzed. The first blood sample was collected immediately at the accident site. The following samples were drawn at intervals from 2 hours to daily for 2 weeks. RESULTS: Sixty-one patients survived the observation time. Immediately after trauma, their mean sCD14 level was not different from that of healthy individuals. Two hours later, a pronounced increase of sCD14 was observed and sustained throughout the observation period. Even nonsurvivors showed an increased sCD14 release, but less pronounced. In all patients, plasma LPS levels were elevated during the first 12 hours. CONCLUSIONS: Major trauma caused an increased release of sCD14. This elevation, however, was not correlated to LPS levels or to the severity of trauma (estimated by trauma scores). We found no evidence that sCD14 levels are of prognostic value regarding survival. Furthermore, the release of sCD14 did not occur in an LPS-neutralizing manner, but rendered possible an LPS-independent mechanism. PMID- 9336512 TI - Of balloon axilloscopy and avoidance of iatrogenic injury to the long thoracic nerve. AB - BACKGROUND: Laparoscopic removal of axillary lymph nodes is possible and affords an excellent view of all structures, allowing preservation of vessels and nerves. The technique uses pediatric trocars and a lifting device to maintain the newly created axillary space. OBJECTIVE: To prove that a newly developed technique of balloon axilloscopy can be performed using only one 10-mm and two 5-mm standard trocars and constant carbon dioxide flow to preserve the axillary space and that preservation of all nerves and vessels is possible with this approach. DESIGN: Prospective study on 4 fresh-frozen human cadavers and 7 live porcine models. SETTING: A hospital department of minimal surgery access and a university department of anatomy. RESULTS: The balloon dissection consistently revealed and preserved the nerves and vessels, and exposure and dissection of the first rib could similarly be accomplished. An alternative route to the apex of the axilla has been developed--between the pectoralis minor and pectoralis major muscles- after their careful separation. The axillary content of surgical interest (lymph nodes) is easily separated from the other anatomical elements and is simply dissected under complete visualization and preservation of all vital axillary structures. CONCLUSIONS: Balloon axilloscopy was easy to perform, provided the surgeon with constant visualization of vital anatomical structures, and allowed easy separation and dissection of the axillary lymph nodes and the first rib. As a technical aid prior to a conventional axillary dissection, or as part of a pure endoscopic procedure in the axilla, balloon axilloscopy is 100% reliable in identifying the long thoracic nerve and moving it out of the way, separating the lymph nodes from it and from the intercostobrachial nerve and axillary vein and artery, rendering the whole dissection process safer for both the surgeon and the patient. PMID- 9336513 TI - Surgery in elderly patients in Mexico. Portal hypertension surgery as an example. AB - Although people older than 65 years represent less than 5% of Mexico's registered population, medical care for elderly patients requires a multidisciplinary approach. In our academic university hospital, they are managed by a team of specialists. As an example of this approach, we evaluated the surgical treatment of bleeding portal hypertension in a highly selected elderly population. A retrospective study was done reviewing the files of 25 patients older than 65 years. All had good liver function (Child-Pugh class A and B) and had undergone elective surgery. Sixteen of them were women. The mean age was 68.8 years (age range, 65-76 years), and most had a diagnosis of cirrhosis. All patients were treated with portal blood flow-preserving procedures (selective shunts or Sugiura Futagawa procedures). The operative mortality was 8%. Eight later deaths were recorded, with a mean follow-up of 25 months (range, 2-110 months). Survival (Kaplan-Meier) was 87% at 12 months, 54% at 60 months, and 45% at 110 months. Two rebleeding incidents were recorded as well as 3 cases of postoperative encephalopathy. We concluded that well-selected elderly patients, undergoing elective surgery with portal blood flow-preserving procedures, have a good postoperative outcome. PMID- 9336514 TI - Treatment of acute cholangitis due to choledocholithiasis in elderly and younger patients. AB - OBJECTIVE: To evaluate management strategies for acute cholangitis in elderly patients (age, > or = 80 years). DESIGN: Nonrandomized control trial. SETTING: A university hospital. PATIENTS: Patients (n = 191) who underwent urgent biliary drainage for acute cholangitis due to choledocholithiasis. Thirty-seven patients were elderly, and 154 were younger (age, < 80 years). INTERVENTIONS: Surgical (8 elderly and 48 younger patients), percutaneous transhepatic (11 elderly and 47 younger patients), or endoscopic drainage (18 elderly and 59 younger patients). MAIN OUTCOME MEASURES: Clinical features of acute cholangitis and outcomes of biliary drainage. RESULTS: The elderly patients had higher incidences of septic shock or mental confusion (acute severe cholangitis)(43.2%) and concomitant diseases (81.1%) than the younger patients (25.3% and 42.9%, respectively). The elderly patients had significantly greater morbidity (37.8%) and mortality (10.8%), compared with the younger patients (16.9% and 3.2%, respectively). Mortality was 18.8% in elderly patients with severe cholangitis and 4.8% in those with nonsevere cholangitis. In the elderly patients, endoscopic drainage yielded lower morbidity (16.7%) and mortality (5.6%) than surgical (87.5% and 25.0%, respectively) and percutaneous drainage (36.4% and 9.1%, respectively). No complications occurred after endoscopic nasobiliary drainage without sphincterotomy. CONCLUSIONS: Elderly patients with acute cholangitis have high incidence of severe disease and concomitant medical problems. They should undergo endoscopic biliary drainage, especially nasobiliary drainage without sphincterotomy, because of its safety and effectiveness. PMID- 9336515 TI - Effect of the Asymptomatic Carotid Atherosclerosis Study on carotid endarterectomy in Veterans Affairs medical centers. AB - OBJECTIVE: To examine the effect of the Asymptomatic Carotid Atherosclerosis Study on the volume of carotid endarterectomies (CEAs) performed in Veterans Affairs medical centers. DESIGN: The data were retrospectively extracted from the Veterans Affairs Patient Treatment File for all patients undergoing CEA using the International Classification of Diseases, Ninth Revision, Clinical Modification procedural code 38.12. Data were classified into patient management categories to identify complications and to quantify the severity of illnesses and comorbidities. SETTING: All 172 US Veterans Affairs medical centers. PATIENTS: Veterans undergoing CEA during fiscal years 1993 through 1995. MAIN OUTCOME MEASURES: Procedural volume, mortality, and morbidity. RESULTS: There was a 43.4% increase in the volume of CEAs performed in fiscal year 1995 despite a 4.6% decrease in the served inpatients and an 8.8% decrease in the inpatient surgical procedures. The monthly volume of CEAs increased (P < .001, r2 = 0.78) at the onset of the fiscal year (October 1994) immediately after the Asymptomatic Carotid Atherosclerosis Study clinical advisory. The volume of CEAs increased in every region of the country for all nonpsychiatric hospital classifications and for almost every surgeon subspecialty. Despite the increased volume, the operative mortality rate, the International Classification of Diseases, Ninth Revision, Clinical Modification--and patient management categories--based complication rates, and the patients' comorbidity and severity of illness indexes all remained unchanged. CONCLUSION: The dramatic increase in CEAs following the Asymptomatic Carotid Atherosclerosis Study clinical advisory suggests that the conclusions of the trial have been accepted by the medical community throughout the Veterans Affairs medical centers. PMID- 9336516 TI - Laparoscopic inferior and superior lumbar hernia repair. AB - Lumbar hernias are rare defects that involve the extrusion of retroperitoneal fat or viscera through a weakness in the posterior abdominal wall. Repairing these hernias is often difficult because of the weakness of the surrounding structures. Techniques for reconstruction usually include an incision from the 12th rib to the iliac crest with mobilization of local flaps or onlay fascial flaps or the use of prosthetic mesh. Contemporary reports have advocated extensive retroperitoneal dissection with the placement of permanent mesh extraperitoneally. We have recently repaired an extensive, primary lumbar hernia laparoscopically, securing the mesh to the 12th rib superiorly, iliac crest inferiorly, erector spinae fascia medially, and external oblique fascia laterally. The patient resumed normal activities in less than 2 weeks; 4 months postoperatively, he seems to have a solid repair. To our knowledge, this is the first report of this technique. PMID- 9336517 TI - 'Domino' liver transplantation combined with multivisceral transplantation. AB - Transplantation of the liver contemporaneously with another organ from the same donor is thought to confer an immunologic advantage. The latter is particularly desirable in intestinal transplantation because of the propensity of the intestinal graft to early and late rejections and because in some cases it may facilitate the operation. In clinical practice, shortage of liver grafts constrains liver transplantation to cases in which there is coexisting end stage liver disease. PMID- 9336518 TI - Developing standards for the treatment of rectal cancer. PMID- 9336519 TI - The role of axillary node dissection in invasive breast cancer. PMID- 9336520 TI - Case 17. Presentation. High grade small-bowel obstruction with massive pneumoperitoneum. PMID- 9336521 TI - Complete femoral osteolysis associated with pathologic fracture. PMID- 9336522 TI - Fat embolism syndrome. AB - Fat embolism syndrome, an important contributor to the development of acute respiratory distress syndrome, has been associated with both traumatic and nontraumatic disorders. Fat embolization after long bone trauma is probably common as a subclinical event. Fat emboli can deform and pass through the lungs, resulting in systemic embolization, most commonly to the brain and kidneys. The diagnosis of fat embolism syndrome is based on the patient's history, supported by clinical-signs of pulmonary, cerebral and cutaneous dysfunction and confirmed by the demonstration of arterial hypoxemia in the absence of other disorders. Treatment of fat embolism syndrome consists of general supportive measures, including splinting, maintenance of fluid and electrolyte balance and the administration of oxygen. Endotracheal intubation and mechanical ventilatory assistance can be indicated. The role of corticosteroids remains controversial. Early stabilization of long bone fractures has been shown to decrease the incidence of pulmonary complications. Clinical and experimental studies suggest that the exact method of fracture fixation plays a minor role in the development of pulmonary dysfunction. As more is learned about the specifics of the various triggers for the development of fat embolism syndrome, it is hoped that the prospect of more specific therapy for the prevention and treatment of this disorder will become a reality. PMID- 9336523 TI - Primary breast cancer in the elderly. AB - OBJECTIVES: With respect to breast cancer in the elderly, to define "old" in the context of comorbidity and physiologic rather than chronologic age. In addition, after discussion of factors influencing decisions regarding screening, stage at presentation and treatment decisions, to present an approach to the treatment of primary breast cancer in the elderly, taking into account quality of life, expected outcomes and cost-effectiveness. DATA SOURCES: A review of the medical literature from 1980 to 1996, using the MEDLINE database and 2 relevant studies from The Henrietta Banting Breast Centre Research Programme at Women's College Hospital, Toronto. STUDY SELECTION: A large number of breast cancer studies that might provide a better understanding of primary breast cancer in the elderly. DATA SYNTHESIS: The studies reviewed demonstrated that the annual incidence of breast cancer increases with age, along with a longer life expectancy for women. There appears to be a delay in presentation for elderly women with breast cancer, related in part to patient and physician knowledge. Biennial mammography and physical examination are effective in women aged 50 to 74 years, but compliance with screening recommendations decreases with age. Although treatment goals are the same for women of all ages, most treatment decisions are based on studies that seldom include women over 65 years of age. Physicians tend to underestimate life expectancy and older women are less likely to seek information. Breast conserving surgery, partial mastectomy and even axillary dissection can be carried out under local anesthesia with little physiologic disturbance, but unless axillary dissection is required to make a treatment decision, it may be foregone in clinically node-negative elderly women. The role of adjuvant radiotherapy in the elderly is not yet well established; tamoxifen is the usual adjuvant systemic therapy given to older women. For those who are truly infirm, tamoxifen alone can be considered. Studies to date do not clarify whether breast cancer in older women runs a more or less favourable course. However, locoregional recurrence appears to decrease with age. Deaths from competing causes are a confounding issue. CONCLUSIONS: It is imperative to develop a coherent strategy for the treatment of primary breast cancer in the elderly that takes into account functional status and quality of life. Clinical trials must include older women and there must be good clinical trials designed specifically for older women. PMID- 9336524 TI - Symposium on rectal cancer: 2. Local recurrence after surgery for rectal cancer. AB - Local recurrence is a serious complication in patients with rectal cancer because of the frequency with which it occurs, its impact on quality of life and the fact that treatment is rarely successful. Although local recurrence rates varying from 4% to 51% have been reported, recent series have reported rates of less than 10%. Various factors may affect the rate of local recurrence, including the stage and location of the tumour. Other prognostic factors may be of importance, but it is controversial whether they are independent risk factors. Finally, there is mounting evidence that the local recurrence rate varies with the surgeon. Whether this is due to the surgical technique or surgical expertise is not clear, but randomized controlled trials addressing the issue of extent of resection are indicated in order to optimize surgical results. PMID- 9336525 TI - Symposium on rectal cancer: 3. The case for adjuvant radiotherapy for rectal cancer. AB - Adjuvant radiotherapy for rectal cancer is intended to eradicate subclinical deposits of cancer cells not removed at surgery. These residual cells are found most commonly at the resection margin of the primary tumour and in transected cancer-bearing lymphatics or vessels. Refinements in surgical technique have been associated with a reduction in the risk of pelvic recurrence in some nonrandomized series. However, clinical trials have shown that the combinations of radiotherapy and chemotherapy, and in some instances radiotherapy alone, reduce the risk of recurrence and may improve survival rates compared with those of surgery alone. It is premature to consider that adjuvant pelvic radiotherapy is unnecessary. PMID- 9336527 TI - Prevalence of heterotopic ossification in cemented versus noncemented total hip joint replacement in patients with osteoarthrosis: a randomized clinical trial. AB - OBJECTIVE: To determine the prevalence of heterotopic bone formation in cemented versus noncemented total hip joint replacement. DESIGN: A prospective randomized controlled trial. Follow-up ranged from 2 to 6 years (mean 4 years). SETTING: A university hospital. PATIENTS: Two hundred and twenty-six patients who had primary or secondary osteoarthrosis of the hip were stratified according to type of fixation, surgeon and age. Patients were randomized within strata: 112 received noncemented total hip prostheses and 114 received cemented prostheses. The 2 groups were similar with respect to age and sex. INTERVENTION: Primary total hip arthroplasty. A cemented (methylmethacrylate) or noncemented prosthesis was inserted by a lateral surgical approach. MAIN OUTCOME MEASURE: The Brooker classification was used to grade heterotopic bone formation from postoperative radiographs. RESULTS: Overall, 148 (66%) hips had no heterotopic ossification, 56 (25%) were Brooker class I, 14 (6%) were class II, 8 (3%) were class III and none were class IV. In the noncemented group of patients, 76 (68%) hips had no heterotopic ossification, 25 (22%) were Brooker class I, 7 (6%) were class II, 4 (4%) were class III and none were class IV. In the cemented group of patients, 72 (63%) hips had no heterotopic ossification, 31 (27%) hips were Brooker class I, 7 (6%) were class II, 4 (4%) were class III and none were class IV. CONCLUSION: There was no significant difference in the prevalence of heterotopic ossification between cemented and noncemented total hip replacements in patients with osteoarthrosis. PMID- 9336526 TI - Fracture of the proximal phalanx of the little finger in children: a classification and a method to measure the deformity. AB - OBJECTIVE: To develop an improved method for measuring the deformity caused by fracture of the proximal end of the proximal phalanx of the little finger in children. DESIGN: A prospective case study. SETTING: Regional hospitals with an orthopedic service. PATIENTS: Forty-two children with a proximal phalangeal fracture of the little finger and 42 children without a phalangeal fracture, who acted as a control. The type of deformity resulting from the fracture was noted, and the angle of deformity was measured. Rotational deformities were measured clinically in all patients and angulation deformities were measured from radiographs. The deformities were graded and classified. MAIN OUTCOME MEASURES: Measurements of the fracture deformity before and after manipulation. RESULTS: There were 38 ulnar angulation deformities, 26 dorsal angulation deformities, 10 ulnar rotation deformities, 3 palmar angulation deformities, 2 radial angulation deformities and 1 radial rotation deformity. The deformities could be graded into 6 different types. CONCLUSION: The measurements of deformity made it possible to describe and classify isolated deformities and combinations of various deformities. PMID- 9336528 TI - Axillary node dissection in patients with breast cancer diagnosed through the Ontario Breast Screening Program: a need for minimally invasive techniques. AB - OBJECTIVE: To determine the role of axillary node dissection by studying patient and tumour characteristics of invasive breast cancer through the Ontario Breast Screening Program (OBSP). DESIGN: A retrospective evaluation. SETTING: The London, Ont., branch of the OBSP. PATIENTS: Three groups of women seen were studied: 50 women with screen-detected breast cancers, which were palpable and detected by the nurse-examiner, 62 women with occult screen-detected breast cancers and 353 age matched women with invasive breast cancer from the LRCC prospective database, who served as controls. MAIN OUTCOME MEASURE: The proportion of involved axillary nodes within the 3 groups based on patient and tumour characteristics. RESULTS: Twenty-five (22.3%) of the 112 women had screen detected tumours less than 1 cm in dimension, but only 1 had an involved axillary node. Twelve (19%) of the 62 women with occult screen-detected tumours had involved lymph nodes compared with 17 (34%) of the 50 women with palpable screen detected cancers (NS). In the control group tumour dimension less than 1 cm versus 1 cm or larger had a marked effect on the probability of axillary node involvement (12.5% v. 40.7%, p = 0.001). In the palpable screen-detected group 3 times as many women with outer quadrant or central lesions had involved nodes as those with inner quadrant lesions (38% v. 12%) and for those with a family history of breast cancer almost twice as many had involved axillary nodes. In occult screen-detected patients there was more nodal involvement in patients aged 60 years or less than in those older than 60 years (35% v. 10%, p = 0.042); only 4 of 41 patients older than 60 years had involved nodes at surgery. A significant difference in nodal involvement was found with respect to high or intermediate grade versus low grade lesions in the occult group (44% v. 12%, p = 0.033). In the control group, tumour grade (intermediate and high [45.3%] v. low [20.0%]) and hormone replacement therapy (HRT) (current or recent use [56.5%] v. no use [34.5%]) were significant findings (p < 0.001 and p = 0.005 respectively). CONCLUSIONS: Women older than 60 years with tumours smaller than 1 cm had a low probability of nodal positivity (0% to 8.7%), but there is insufficient information in this group to give a 95% or better prediction of nodal status at the time of surgery. Studies of minimally invasive techniques such as sentinel node biopsy are needed in this group to minimize surgical morbidity in these women who, as a result of early diagnosis, have an excellent long-term outlook. PMID- 9336529 TI - Hip fracture surgery in Nova Scotia: a comparison of treatment provided by "generalist" general surgeons and orthopedic surgeons. AB - OBJECTIVE: To determine quality of hip fracture services provided by "generalist" general surgeons (generalists) in Nova Scotia. DESIGN: Chart review and postoperative, blinded, random-ordered radiologic analysis. SETTING: Three community hospitals and 1 tertiary care hospital in Nova Scotia. PARTICIPANTS: Seven generalists who performed 120 hip fracture repairs and 7 orthopedic surgeons (specialists) who performed 135 hip fracture repairs. OUTCOME MEASURES: Patient demographics, preoperative, perioperative, postoperative and discharge information, technical quality of reduction as determined through postoperative radiologic assessment. RESULTS: There were no differences between patients treated by generalists and those treated by specialists with respect to age, sex, American Society of Anesthesiologists' class, level of function and fracture type. Intraoperatively, the patient groups were similar with respect to type of anesthesia, use of antibiotics, number of transfusions and surgical complications. Significant differences were noted in length of operation (54.4 v. 41.1 minutes), use of C-arm imaging (6.7% v. 85.9%) and management of Garden classes 1 and 2 subcapital fractures. Postoperatively, the 2 groups had similar numbers of medical complications, wound complications, reoperations, readmissions and deaths, and a similar level of function on discharge. Significant differences included the number of intensive care unit admissions (5.8% v. 15.6%) and length of stay there (5.7 v. 2.8 days) and of postoperative stay (14.5 v. 10.7 days). The assessment of radiographs did not demonstrate any significant difference in the quality of reduction. CONCLUSION: In Nova Scotia the outcomes of hip fracture surgery performed by generalists are comparable to those performed by specialists. PMID- 9336530 TI - Carcinoma of the stomach: an unusual pattern of metastasis. PMID- 9336551 TI - Milestones. The heritage of Army dietetics. PMID- 9336552 TI - ADA's medical nutrition therapy campaign reaches milestone. PMID- 9336553 TI - New trends in weight management. PMID- 9336554 TI - Computerized patient record: are we prepared for our future practice? AB - OBJECTIVE: To survey members of The American Dietetic Association (ADA) regarding care documentation systems, computerization of patient care records, and factors to be considered in developing a documentation system compatible with a computer based patient record. DESIGN: The survey instrument was developed in conjunction with a survey consultant/statistician, then mailed to the study sample. SUBJECTS/SETTING: The sample of 500 was drawn from three ADA dietetic practice groups expected to include a high percentage of clinical practitioners. STATISTICAL ANALYSIS PERFORMED: Basic frequency displays were used on all questionnaire items. Pearson correlation coefficients were used among numeric variables, and oneway analysis of variance was used for categoric variables with quantitative variables. RESULTS: A total of 171 usable surveys were returned (34%), primarily from dietitians working in an acute-care inpatient environment. The SOAP format (subjective, objective, assessment, and plan) was used by 60% of respondents to document nutrition assessments, although a number of other documentation formats were reported. Most commonly used data in nutrition decision making were medical diagnosis, diet order, anthropometric data, and laboratory values. Most commonly used outcomes measures included laboratory values, tolerance of the nutrition regimen, weight changes, and intake changes. Only 15% of respondents reported that they currently used a computerized patient record. Ninety-three percent of respondents favored standardized nutrition diagnoses, and 95% believed standardized nutrition interventions would prove useful. APPLICATIONS/CONCLUSIONS: We recommend that dietitians evaluate, standardize, and streamline their documentation to prepare for implementation of computerized systems. The diagnoses and interventions presented in this study could be a starting point. PMID- 9336555 TI - Practice points: translating research into practice. Implementing computerized patient records: a success story. PMID- 9336556 TI - Comparative evaluation of body composition in medically stable elderly. AB - OBJECTIVE: Accurate assessment is needed to identify the nutritional status of elderly persons. Anthropometric data were collected to describe body composition of the sample, including blacks and whites aged 55 to 89 years, and to explore the usefulness of several methods of body composition measurement. DESIGN: Baseline measurements were made as part of a longitudinal study. Body composition variables, particularly lean and lean-to-height measures, were used. SUBJECTS/SETTING: One hundred twenty-nine free-living medically stable elderly at senior community centers were self-selected into the study. STATISTICAL ANALYSIS: Descriptive statistics were generated for all variable by gender, race, and age (< 65, 65 to 74, and > or = 75 years). Actual height was correlated with estimated height using published equations. Analysis of variance revealed the effect of gender, race, or age on outcome variables. RESULTS: White women, black women, and white men made up 54%, 23%, and 23% of the sample, respectively. Most gender differences were expected. Black women had greater weight, body mass index, arm muscle circumference, and ratio of lean to height but lower percent lean body mass than white women. Subjects over 75 years old were shorter, lighter, and had lower ratio of lean to height. Gender and age had the greatest effect on ratio of lean to height. CONCLUSIONS/APPLICATIONS: A lean-to-height index appears to be a useful tool for tracking the status of lean mass in the elderly. Knee height may be especially useful because it, unlike stature, does not decrease with age. Furthermore, some published equations for estimating stature from knee height need adjustment, specifically for elderly black women. PMID- 9336557 TI - No ergogenic effects of ginseng (Panax ginseng C.A. Meyer) during graded maximal aerobic exercise. AB - OBJECTIVE: To assess the effects of chronic supplementation with two different dosages of Panax ginseng C.A. Meyer on physiologic and psychological responses during graded maximal aerobic exercise. DESIGN: Randomized, double-blind, placebo controlled trial. SUBJECTS: Thirty-six healthy men consuming an otherwise supplement-free diet who maintained their usual activity level. INTERVENTION: A standardized P ginseng C.A. Meyer concentrate (G115) was added to the normal diet of study participants at a dosage level of either 200 or 400 mg/day, where 100 mg of the preparation is equivalent to 500 mg P gingseng root. MAIN OUTCOME MEASURES: Submaximal and maximal aerobic exercise responses before and after an 8 week trial intervention. STATISTICAL ANALYSES PERFORMED: Analysis of variance. RESULTS: Thirty-one subjects completed the study. Supplementation with ginseng had no effect on the following physiologic and psychological parameters: oxygen consumption (mL/kg per minute), respiratory exchange ratio, minute ventilation (L/min), blood lactic acid concentration (mmol/L), heart rate (beats/min), and perceived exertion (P > .05). CONCLUSIONS: Our data in healthy men do not offer support for claims that P ginseng C.A. Meyer is an ergogenic aid to improve submaximal and maximal aerobic exercise performance. PMID- 9336559 TI - Review of self-efficacy and locus of control for nutrition- and health-related behavior. AB - This article reviews several cognitive predictors of health- and diet-related behaviors commonly used in theories and models of nutrition and health behavior change. Constructs such as self-efficacy, self-esteem, outcome expectancies, health value, and locus of control are examined. Self-efficacy has repeatedly been a good predictor of health behavior, sometimes explaining more than 50% of variability. Research on locus of control and other predictive factors has been less conclusive. The take-home message is threefold: (a) task specificity of self efficacy and domain specificity of locus of control are crucial for unraveling their effects on behavior; (b) careful segmentation of different population groups under study may explain the inconsistencies in previous research; and (c) especially when studying dietary behavior, these predictors of behavior change should not be used alone or in place of one another but should be used simultaneously to explain complex food and diet-related behaviors. We recommend that nutritionists systematically integrate available theories and models and explore new areas for studying human behavior, such as sociology and anthropology, to form a more powerful, comprehensive model for behavior change. PMID- 9336560 TI - The Solution Method: 2-year trends in weight, blood pressure, exercise, depression, and functioning of adults trained in development skills. AB - This study describes changes observed during a 2-year period in participants enrolled in The Solution Method, a developmental skills training program for adult weight management. This intervention is the adult application of a model of treatment previously used only in the management of pediatric obesity (The Shapedown Program). Developmental skills training integrates understandings and methods from developmental, family systems, biomedical, genetic, and behavioral theories of the etiology of obesity. Twenty-two subjects (mean age = 43.4 +/- 8.5 years and mean body mass index = 33.1 +/- 5.3) completed a group intervention based on this method, which was conducted by a registered dietitian and a mental health professional. Questionnaire responses indicated the extent to which their weight was a medical and/ or psychosocial risk. Subjects attended 2-hour weekly sessions for an average of 18 weeks during which they were trained in six developmental skills: strong nurturing, effective limits, body pride, good health, balanced eating, and mastery living. Data, which were collected at the beginning of treatment and at 3, 6, 12, and 24 months, included weight, blood pressure, 7-day exercise recalls, and responses to depression and functioning (psychosocial, vocational, and economic) questionnaires. Participants' weights decreased throughout the 2-year period of the study: mean weight change was -4.2 kg (3 months), -6.0 kg (6 months), -7.0 kg (12 months), and -7.9 kg (24 months). In addition, compared with baseline values, systolic and diastolic blood pressure, exercise, and depression improved throughout the study period. These improvements were statistically significant at 24 months for weight (P < .01), systolic blood pressure (P < .02), diastolic blood pressure (P < .001), and exercise (P < .001); the results were not statistically significant for depression. Most participants reported improvement in a broad range of aspects of functioning. We conclude that this application of developmental skills training for adult weight management may produce significant long-term beneficial effects. PMID- 9336561 TI - Database and quick methods of assessing typical dietary fiber intakes using data for 228 commonly consumed foods. AB - To promote assessment of dietary fiber intakes in clinical settings, we established two objectives for this study: to provide a detailed database in grams per serving of fiber content and polymer composition for most fiber sources in the US diet, and to develop a quick method for estimating total fiber intakes. Data for 342 foods were condensed to 228 foods by combining similar foods. The comprehensive database developed includes pectin, hemicelluloses, and beta-glucan contents of the soluble and insoluble fractions of fiber and the cellulose and Klason lignin contents of the insoluble fiber. Three fourths of the 228 foods contained 2.0 g fiber per serving or less; only 10% contained more than 3.0 g per serving. The quick method consists of multiplying the number of servings in each food group by the mean total dietary fiber content of foods in that group: 1.5 g for fruits (n = 43), 1.5 g for vegetables (n = 68), 1.0 g for refined grains (n = 80), and 2.5 g for whole grains (n = 13). Actual fiber values from the database should be used in the quick method if foodstuffs concentrated from grains, legumes, and nuts and seeds are consumed. Sample menus demonstrate that quick assessment of total fiber intake yielded results similar to the sum of individual values from the database. PMID- 9336558 TI - Twelve weeks of endurance exercise training does not affect iron status measures in women. AB - OBJECTIVE: This study examined the effects of 12 weeks of endurance exercise training on iron status measures in previously inactive women and compared the effects of weight-bearing endurance exercise training and non-weight-bearing endurance exercise on iron status measures. DESIGN: Randomized, experimental study. SUBJECTS: Thirty-one healthy, inactive women (aged 23 to 43 years) with apparently normal iron stores (serum ferritin concentration > or = 20 micrograms/L) were recruited from the local area by newspaper advertisements and campus mailings. Twenty-one subjects completed the study (mean +/- standard deviation for age = 32 +/- 5 years, for body mass index = 23.1 +/- 4.9, and for maximum oxygen consumption [VO2max] = 33.8 +/- 6.3 mL/kg per minute). INTERVENTION: Subjects were randomly assigned to one of three groups: an inactive control group, a walking/running group, or a cycling group. Subjects in the two exercise groups trained three to four times per week at 80% VO2max for 12 weeks. Exercise training sessions were monitored and energy expenditure increased from 150 kcal per session (week 1) to 375 kcal per session (weeks 9 to 12). Subjects in the inactive control group were instructed to maintain their normal activity patterns for the duration of the study. All subjects were instructed to maintain their normal dietary habits. MAIN OUTCOME MEASURES: Serum ferritin concentration, serum iron concentration, percentage saturation of transferrin, total iron binding capacity, serum haptoglobin concentration, and other selected hematologic variables were measured at baseline and at weeks 2, 4, 8, and 12. STATISTICAL ANALYSES: Repeated measures analysis of variance was used to examine group x time interactions in changes in iron status measures. Statistical significance was reached at P < .05. RESULTS: Analysis of variance indicated that serum ferritin concentration did not change significantly (P = .59) during the 12 weeks in the walking/running group (mean +/- standard deviation from 41.28 +/- 14.22 to 27.41 +/- 9.74 micrograms/L) or the cycling group (from 65.81 +/- 37.62 to 41.06 +/- 26.38 micrograms/L) compared with the control group (from 47.55 +/- 15.87 to 31.56 +/- 10.57 micrograms/L). Values for serum iron, total iron-binding capacity, percentage saturation of transferrin, and haptoglobin also did not change significantly (P > .30) in the walking/running or cycling groups compared with the control group. APPLICATIONS/CONCLUSIONS: Results of this study suggest that participation in 12 weeks of moderate-intensity endurance exercise training (walking/running or cycling) is not associated with negative effects on selected measures of iron status in healthy, previously untrained women with normal iron stores (serum ferritin > or = 20 micrograms/L). PMID- 9336562 TI - Health care foodservice directors' perception of the importance of value-added services offered by foodservice distributors. PMID- 9336563 TI - Nutrition via jejunostomy in refractory hyperemesis gravidarum: a case report. PMID- 9336564 TI - Position of the American Dietetic Association: health implications of dietary fiber. PMID- 9336565 TI - Jackie Krick wins Huddleson Award. PMID- 9336566 TI - Research competencies in the dietetics curricula. AB - Investment in dietetics research has the potential to improve general health; increase work performance and learning capacity; extend disease-free life; reduce birth defects, infections, and chronic diseases; and decrease health care costs. Opportunities exist for research in the areas of solid waste management, global environment, health care reform, and foodservice systems management. Research efforts can focus on cost-effectiveness and medical efficacy of dietary services to improve third-party reimbursement. Educators have a stake in creating the future by conducting research, teaching research to dietetic students, and investigating the effectiveness of education. PMID- 9336567 TI - Pediatric nutrition assessment: identifying children at risk. AB - Nutrition services are important in the prevention of disabilities as well as in the treatment and/or habilitation of children with chronic illness. Level 1 nutrition care requires some basic knowledge of nutrition to screen for nutritional risk factors, knowledge of and access to referral systems for children identified to be at risk, and ability to use general nutrition education materials. Level 2 involves individualized nutrition assessment and intervention for problems such as anemia, chronic constipation, low- or high-calorie diets, feeding problems, and growth monitoring. Level 3 nutrition services are for children with identified disabilities such as cystic fibrosis, diabetes, and metabolic disorders that require specific complex nutrition interventions. The five major components of assessment of nutritional status in children are: anthropometric, clinical, biochemical, dietary, and feeding skill development. PMID- 9336568 TI - Empower children to develop healthful eating habits. AB - Controlling a child's eating habits is counterproductive. By allowing children to make decisions about what and how much to eat, parents empower children to self regulate their eating. The parent's role is to offer a variety of healthful foods, oversee the planning and assembly of meals, and set the schedule for meals and snacks. The child's responsibility is to decide what, how much, and even whether to eat. PMID- 9336569 TI - School-based health clinics: the role for nutrition. AB - There are three primary areas for nutrition intervention in schools: direct nutrition care in a school-based health clinic, school meals, and nutrition education in the classroom and of school staff. Dietitians can positively affect students' nutrition health by providing nutrition guidance in the clinic and by creating a positive nutrition environment in the school through increased nutrition education in the classroom and healthful, tasty meals in the cafeteria. PMID- 9336570 TI - Elder insecurities: poverty, hunger, and malnutrition. AB - Between 8% and 16% (2.5 to 4.9 million) of the elder population have experienced food insecurity within a 6-month period. Federal programs to combat food insecurity reach only one-third of needy elders. While hunger and poverty are linked directly to malnutrition, the multifaceted nature of elderly malnutrition cuts across all economic, racial, and ethnic groups. Malnourished patients experience 2 to 20 times more complications, have up to 100% longer hospital stays, and compile hospital costs $2,000 to $10,000 higher per stay. Dietitians can advocate routine nutrition screening to target elders at highest risk and lobby for expansion of appropriate nutrition services in home, community, and institutional settings. PMID- 9336571 TI - Oral health screening guidelines for nondental health care providers. AB - Clues identified in oral health screening can be associated with medical conditions, medical treatments, and overall health status. Thorough oral health screening involves not only talking with the patient, but touching and exploring the mouth. Oral health screening expands the role of dietitians in improving the nutritional and dietary practices of clients. PMID- 9336572 TI - Dietary studies in the joint US-Russian space program. AB - Metabolic experiments in the joint US-Russian space program involve analysis of food records, which include weighed foods, stable-isotope turnover, and biochemical samples collected before, during, and after the flights. This article describes the methods of monitoring dietary intake for this program. PMID- 9336573 TI - Building your business--setting your fees: a cost-based approach. AB - Business costs include salary, benefits, self-employment tax, and business and office expenses. Salable time does not include holidays, sick or personal days, and vacations. According to fee-setting experts, most self-employed people engage in revenue-generating activities 50% to 75% of their working hours. Fair prices reflect the cost of your business or service, including all your overhead. PMID- 9336574 TI - Charting by exception: a solution to the challenge of the 1996 JCAHO's nutrition care standards. AB - With charting by exception, only significant findings or exceptions to the norms are documented in script by the nursing staff. Upon admission, patients are screened for nutrition risk or need for nutrition education by nursing staff using an interdisciplinary patient database. Patients with predetermined criteria indicating nutritional risk or with nutrition education needs are referred to nutrition services. Using specified criteria, the clinical nutrition staff assign referred patients to a level of care. Nutrition care or education is provided and documented within 48 hours. Documentation varies with the level of nutrition care. PMID- 9336575 TI - Do you know what a dietetic technician can do? A focus on clinical technicians and their expanded roles and responsibilities. AB - Dietetic technicians, registered, play a key role in providing quality, cost effective client care and foodservice management and are guided by the mission statement of the American Dietetic Association to promote optimal nutrition, health, and well-being. Many organizations use dietetic technicians to complete the screening and/or basic level assessments. The dietetic technician also can assist with patient monitoring and complete such tasks as calculating and documenting calorie counts, providing basic diet instruction, monitoring diet consumption and compliance, and determining tolerance of nutritional supplements. Since the last role delineation study was conducted, dietetic technicians have expanded into many different work settings, and practice roles have changed accordingly. PMID- 9336576 TI - Cyberspace 101: taking a ride on the information superhighway. AB - All you need to explore cyberspace is a computer, a modem, a phone line, and a local "on-ramp" to the infohighway. A litserv is an interactive mailing list that distributes information to a large number of people at the same time. Once you subscribe, you receive copies of all messages sent into listserv and have the opportunity to post questions and comments for other subscribers. Dietetics Online: A Network of Dietetic/Nutrition Professionals offers a range of cutting edge services. Online marketing can reach a potentially larger audience for a fraction of the cost of traditional means and expand your business geographically. PMID- 9336577 TI - Overcoming resistance to organizational change. AB - Resistance to assertive organization change is inevitable because people are asked to reexamine and modify their behavior, which breeds resistance. Resistance serves to maintain equilibrium until the reasons for change are both conscious and compelling. Instead of accepting people's feelings as excuses, persistently push for what you know needs to happen in the face of today's harsh realities. Provide clarity, time, support, and the stability of a persistent message. PMID- 9336578 TI - Foodservice design--planning for the future. AB - An independent foodservice consultant performs the design functions and puts the specifications and plans out for a competitive bid to dealers. A foodservice equipment dealer will order and install equipment and handle concerns after installation. Planning and forethought on the part of the foodservice director and consultant can assist the food-service designer in coordinating the functions to create an efficient kitchen. PMID- 9336579 TI - Essential fatty acid deficiency in renal failure: can supplements really help? AB - Abnormal fatty acid metabolism may contribute to clinical problems such as itching, abnormal perspiration, susceptibility to infection, delayed wound healing, anemia, and increased hemolysis, as seen in patients with chronic renal failure. A double-blind study of patients on hemodialysis who received either fish oil, olive oil, or safflower oil documented that patients may have increased levels of the proinflammatory prostaglandin PGE2 and that fish oil intervention may decrease these levels, change the fatty acid profile, improve hematocrit levels, and improve patient perception of symptoms of pruritus. PMID- 9336580 TI - Indirect calorimetry: technical aspects. AB - Indirect calorimetry measures oxygen consumption and carbon dioxide production to calculate resting energy expenditure and respiratory quotient. The respiratory quotient can be determined from indirect calorimetry to determine substrate utilization and used to alter the patient's nutrition support regimen. All but one indirect calorimeter manufactured in the United States are open-circuit rather than closed-circuit systems. PMID- 9336581 TI - HIV and medical nutrition therapy. AB - In those who are infected with human immunodeficiency virus, poor nutritional status can result from numerous causes, including anorexia, catabolism, chronic infection, fever, poor nutrient intake, nausea, vomiting, diarrhea, malabsorption, metabolic disturbances, lack of access to food, depression, and side effects of drug, radiation, and chemotherapy treatments. A compromised immune system may not be reversed by any medical treatments at this time, but malnutrition may be prevented and reversed by using current therapies, including medical nutrition therapy that includes nutrition assessment, the development of an individualized nutrition therapy plan, and implementation of the therapy. There is substantial evidence that medical nutrition therapy saves lives, reduces morbidity, improves health outcomes, reduces costs, and shortens hospital stays. PMID- 9336582 TI - Pattern management: a tool for improving blood glucose control with exercise. AB - Pattern management can help the person with type 1 diabetes to understand glycemic response to activity, make reasonable management decisions, and evaluate the effectiveness of adjustments to exercise. Insulin adjustments and carbohydrate supplementation can be used independently or in conjunction to maintain target-range blood glucose levels during activity. To achieve success, the patient must be encouraged to gather data through self-monitoring of blood glucose and diligent recordkeeping, analyze information collected, make careful adjustments, and assess outcomes. PMID- 9336583 TI - The "to feed or not to feed" dilemma. AB - Advance directives can promote and improve communication between the patient and the provider and ideally safeguard the resident's interests by directing medical care. Participation on the facility's ethics committee allows the dietitian to become involved in the legal and ethical issues regarding feeding and to promote the use of specific advance directives for nutrition and hydration. Consider the goals of patient care carefully and assure that they are individualized and patient-centered. Continually reassess nutritional status and collaborate with the health care team in recommending interventions and care plans for each case. PMID- 9336584 TI - Nutritional needs of the person with Alzheimer's disease: practical approaches to quality care. AB - Intellectual losses in Alzheimer's disease include ever-decreasing attention span, lack of judgment, compromised logic and reasoning skills, and lack of ability to plan and organize complex tasks. Patients have great difficulty staying focused on the task of eating. Extensive damage to the parietal, temporal, and occipital lobes causes altered processing of sensory input, resulting in distorted perceptions of the world around them. Many so-called "behaviors" are the direct result of clients' responses to their perceived environment. The ability to use utensils deteriorates because of both cognitive and motor losses. These patients commonly have slow, progressive weight loss over the span of their illness, despite high-energy diets. Altering the environment, staff approaches, and food items will make patients' lives easier and maintain good nutrition for as long as possible. PMID- 9336585 TI - The use of dietary supplements in the elderly: current issues and recommendations. AB - Research on dietary intake in the elderly shows evidence of both adequate and inadequate nutrient consumption from food. More recent studies have documented inadequate mineral intake from food and confirmed the overall decline in nutrient intake from food as people age. Food is incontrovertibly the best vehicle for nutrient consumption. However, some authorities have found reason to recommend a daily multivitamin-mineral for the elderly as a reasonable way to assure adequate micronutrient intake. There appears to be no reason to recommend complete liquid supplements or modular macronutrient supplements to the active free-living elderly population. PMID- 9336586 TI - The role of maternal nutrition in the prevention of birth defects. AB - "Natural" phenomena allow conclusions that maternal dietary deficiencies or excesses can affect development of the human embryo and fetus adversely. The most important finding in recent years has been the relationship between maternal folic acid status and neural tube defects. PMID- 9336587 TI - Cystic fibrosis, nutrition, and the health care team. AB - Because of the multiple systems involved in cystic fibrosis, the variability and chronicity of the disease, and the increased survival of this population, a specialty team of experts for care has evolved. A multidisciplinary approach is essential to assist patients and their families in adjusting to the disease and to optimize treatment interventions. The dietitian is responsible for assessment of nutritional status, including the determination of energy requirements and eating habits, interpretation of anthropometric data, and evaluation of nutritional adequacy. The nutrition care plan forms an integral part of the overall treatment objectives and is reported to other team members as it is devised, implemented, and monitored. A consensus report issued in April 1990 by the Cystic Fibrosis Foundation includes both general nutrition guidelines and detailed recommended treatment standards aimed at providing optimal nutrition care. PMID- 9336588 TI - The ketogenic diet revisited. AB - The ketogenic diet is a high-fat diet that maintains the body's starvation mechanism, with exogenous fat provided for metabolism in lieu of stored fat. Mild dehydration is important to prevent dilution of the level of ketones in circulation at any given time. It is not known why or how ketosis affects seizure activity, so the principles behind the therapy have been developed from years of clinical experience and theoretical assumptions. Dietitians are essential providers of ketosis therapy, but the dietitian must work with a physician who understands the theories behind the therapy and is an active member of the ketosis therapy team. PMID- 9336589 TI - Dietitians: experts about food systems? AB - Dietary advice that promotes optimal nutrition presupposes adequate land for growing food, environmental quality conducive to food production, stable government, a strong economy, and community food security (or a reliable transportation system). Without these conditions, recommendations for optimal nutrition become moot. Our profession will be strengthened as we develop a broad knowledge of our entire food system. PMID- 9336590 TI - Sports drinks: research asks for reevaluation of current recommendations. AB - Results of two clinical studies support the use of a carbohydrate-electrolyte beverage to improve performance of intermittent moderate- to high-intensity exercise, such as soccer and ice hockey. Two other studies documented that such beverages improved performance during steady-state moderate- to high-intensity exercise, although the mechanism by which carbohydrate improved performance in these protocols is yet to be determined. PMID- 9336591 TI - Phytochemicals: guardians of our health. AB - Consuming a diet rich in plant foods will provide a milieu of phytochemicals, nonnutritive substances in plants that possess health-protective benefits. Nuts, whole grains, fruits, and vegetables contain an abundance of phenolic compounds, terpenoids, pigments, and other natural antioxidants that have been associated with protection from and/or treatment of chronic disease such as heart disease, cancer, diabetes, and hypertension as well as other medical conditions. The foods and herbs with the highest anticancer activity include garlic, soybeans, cabbage, ginger, licorice, and the umbelliferous vegetables. Citrus, in addition to providing an ample supply of vitamin C, folic acid, potassium, and pectin, contains a host of active phytochemicals. The phytochemicals in grains reduce the risk of cardiovascular disease and cancer. PMID- 9336592 TI - Taste: the neglected nutritional factor. AB - Scientists now think that every taste bud has some degree of sensitivity to all the primary taste sensations. The brain detects the type of taste by the ratio of stimulation of different taste buds. Many different substances have flavor enhancing capacity, including ingredients that represent the primary tastes. Medications, chronic disorders, and radiation therapy can alter taste perception, resulting in loss of appetite. Individualizing nutrition advice, with consideration of taste, health needs, and personal preferences, is a "signature dish" of quality dietetics practice. PMID- 9336593 TI - A review of some herbal and related products commonly used in cancer patients. AB - In light of the research in progress on the benefits of various phytochemicals in foods, it appears feasible that the chemical compounds from herbs also could be helpful in prevention or treatment of cancer and other diseases. Prior to 1994, the burden of proving that a product was not harmful was the manufacturer's responsibility. Now the FDA has to prove beyond a doubt that the product is unsafe to remove it from the market. Product standardization is optional at this point, and substitution with less expensive herbs presents a risk. The FDA soon will publish a proposal for good manufacturing practices that was developed by a coalition of supplement organizations to assist in establishing quality control. Dietetics professionals can contribute value in this area by talking with patients, keeping interested physicians informed, and maintaining information on alternative therapies. PMID- 9336594 TI - Pathological gambling, When do social issues become medical issues? PMID- 9336595 TI - Serodiagnosis of Lyme disease. PMID- 9336596 TI - Repetitive strain injury. 1. An overview of the problem and the patients. The Goff Group. AB - Assembly-line workers, house painters, and many others whose activities entail repetitive motions can end up with swelling, pain, and limited movement in the affected muscles. Often, use of the six steps described in this article brings fairly rapid functional improvement and prevents recurrences, with a minimum of medical intervention. In some cases, though, recovery is prolonged or the outcome is unusual. The authors present additional factors to consider in such cases, such as psychosocial concerns, worker fraud, and ergonomic problems. Part 2 of this article, beginning on page 72, details six common repetitive strain injuries. PMID- 9336597 TI - Repetitive strain injury. 2. Diagnostic and treatment tips on six common problems. The Goff Group. AB - Repetitive strain injury is caused by recurrent overuse, resulting in microtrauma to tissues. Local pain and tenderness, weakness, inflammation, and limited function are common findings. Some of the strain injuries seen most often are carpal tunnel syndrome, trigger finger, shoulder impingement syndrome, tennis elbow, thoracic outlet syndrome, and myofascial pain disorders. Often, treatment can be started at the initial visit, after systemic disorders have been ruled out. A vital step is elimination of aggravating factors, such as improper posture, inadequate attention to ergonomic factors at work, and contributory habits (e.g., jaw or hand clenching). Use of simple joint-protection measures can alleviate much of the discomfort. Appropriate self-help strategies used at home may restore flexibility and strength with a minimum of medical intervention, but pain relief must be achieved before patients can be expected to follow through with rehabilitation efforts. Use of ice packs, massage, NSAIDs, or topical pain relief agents is often helpful. Prompt, temporary pain relief can also be achieved with injection of a local anesthetic-corticosteroid mixture. Persistent disability should prompt consideration of psychosocial factors. In addition, fraudulent claims of disability do occur. Although physicians should make every effort to support legitimate claims of work-related injury, they should also be aware of the possibility that activities outside of work (e.g., sports participation, accidental injuries) may be contributing factors. PMID- 9336598 TI - The case of the second tongue. Acute bacterial sialadenitis of the submandibular glands. PMID- 9336599 TI - HIV/AIDS. PMID- 9336600 TI - Antiretroviral therapy for HIV infection. Heartening successes mixed with continuing challenges. AB - Probably nothing in the field of medicine has changed as rapidly over the last decade as antiretroviral (ARV) therapy for HIV infection. New insights into pathogenesis, new tests for measuring virus levels in plasma, and availability of powerful new drugs have combined to transform the care of persons with HIV infection into a field infused with real hope. However, ARV therapy remains complex, expensive, and not universally effective or available. This article focuses on a general approach to the use of ARV agents. PMID- 9336601 TI - Preventing and treating major opportunistic infections in AIDS. What's new and what's still true. AB - New highly active antiretroviral therapies are boosting the blood absolute CD4+ counts of many patients with AIDS and are decreasing the prevalence of AIDS related opportunistic infections. Nevertheless, the prevention, diagnosis, and treatment of opportunistic infections remain important features of management of HIV infection. In recent years, significant advances have been made in the prevention and treatment of opportunistic diseases such as Pneumocystis carinii pneumonia, Cytomegalovirus retinitis, disseminated Mycobacterium avium intracellulare infection, and mucosal candidiasis. Tuberculosis, cryptococcal meningitis, herpes simplex virus infection, shingles, and infectious enteritis also continue to be troublesome. Kaposi's sarcoma may be the newest AIDS-related opportunistic infection to be identified. The immune system effects of highly active antiretroviral therapy are as yet poorly understood. Therefore, an aggressive approach to diagnosis and treatment of opportunistic infections remains mandatory, and patients receiving antiretroviral therapy should continue to adhere to recommendations for prophylaxis against such infections. PMID- 9336602 TI - Primary HIV infection. Early diagnosis and treatment are critical to outcome. AB - It is now apparent that early identification of primary HIV infection is important, because events occurring in early disease may predict how fast a patient progresses to AIDS. However, as Dr Schacker points out, diagnosis is not easy at this stage. This article describes clinical features and laboratory tests to help primary care physicians make a timely diagnosis and discusses current recommendations for therapy. PMID- 9336603 TI - HIV and occupational risk. Evolving ways to protect healthcare workers. AB - HIV has significantly altered the face of healthcare and the lives of virtually everyone in our communities. The risk of transmission, particularly through needlestick injuries, continues to be a major concern for all of us working in healthcare services. Dr Thurn reviews new information about modes of HIV transmission, ways to reduce risks, and guidelines for managing exposures, should they occur. PMID- 9336604 TI - HIV infection in women and children. Special concerns in prevention and care. AB - Unfortunately, HIV infection continues to spread rapidly among women in the United States, and infected women still have a poorer outcome than do infected men. Prevention, early diagnosis, and facilitation of care in women are critical to both their own and their children's health. Fortunately, significant advances continue to be made in prevention of HIV transmission to children. Not only HIV experts but also primary care physicians play an important role in identification and care of HIV-infected women and children. In addition, prevention of HIV infection requires the efforts of professionals throughout the healthcare spectrum. PMID- 9336605 TI - Venous thromboembolism. A contemporary diagnostic and therapeutic approach. AB - Patients with risk factors for VTE and developing signs or symptoms remotely consistent with DVT or PE should undergo formal testing. Ultrasound and IPG have good sensitivity and specificity in symptomatic DVT. V/Q lung scans should be obtained in all patients with suspected PE or proven DVT. A prior clinical assessment of probability, based on risk factors, history, physical examination, chest film, and arterial blood gas analysis, can aid in the interpretation of V/Q scans. Normal scans exclude PE. High-probability scans confirm PE if the clinical probability is at least intermediate. Nondiagnostic scans are common, but diagnosis in such cases can be aided by noninvasive leg studies. Heparin therapy should be started when there is suspicion of VTE. To avoid recurrence, a therapeutic aPTT of 1.5 to 2.5 times the control rate should be achieved as soon as possible after the diagnosis of VTE is confirmed. Thrombolytics are reserved for hemodynamically compromised patients. Warfarin should be administered to achieve an INR of 2.0 to 3.0 and should be continued for at least 3 months in patients with low risk of recurrence and probably for at least 6 months in all other patients. PMID- 9336606 TI - Multiple myeloma. Diagnostic clues in a patient presenting with incapacitating arthralgias. AB - Multiple myeloma is characterized by the production of a monoclonal immunoglobulin, free monoclonal light chains, or both. Although bone pain is the classic presentation, multiple myeloma should be a consideration in differential diagnosis in elderly patients with arthritic complaints if other typical symptoms or laboratory abnormalities, such as anemia, hypercalcemia, and elevated ESR, are present. A combination of radiation therapy and chemotherapy is the usual treatment. PMID- 9336607 TI - Congestive heart failure in elderly patients. The treatment goal is improved quality, not quantity, of life. AB - Primary care physicians who see elderly patients are likely to see cases of congestive heart failure, since this condition is typically the result of long standing hypertension or coronary artery disease. Recognizing the condition in elderly patients may not be easy, though, because clinical signs can be distorted by accompanying symptoms. In this article, the authors discuss pathophysiologic, diagnostic, and pharmacokinetic issues. They also describe therapy with diuretics, angiotensin-converting enzyme inhibitors, and digoxin and outline special considerations in the elderly. PMID- 9336608 TI - Grow old along with me. A look at the new face of aging. PMID- 9336609 TI - Gastric cancer. Update on diagnosis, staging, and therapy. PMID- 9336610 TI - Peripheral neuropathy in patients with chronic renal failure. A treatable source of discomfort and disability. AB - Uremic polyneuropathy occurs in about half of patients undergoing dialysis and is characterized by axonal degeneration with secondary segmental demyelination. Hemodialysis or peritoneal dialysis halts the progress of polyneuropathy but usually does not bring improvement. However, improvement invariably occurs with successful renal transplantation. Mononeuropathies resulting from placement of forearm arteriovenous fistulas are also seen and include the commonly encountered carpal tunnel syndrome and the rare but catastrophic ischemic monomelic neuropathy. The latter constitutes a medical emergency; immediate surgical closure of the fistula is required to avoid severe and permanent neurologic dysfunction. PMID- 9336611 TI - Lateral digital rotation flaps in the treatment of forms of Dupuytren's contracture. 141 cases. AB - Severe contractures of the little finger can be corrected with a rotation flap. It has been used 141 times in a homogeneous series of 522 Dupuytren diseases. This flap brought to the metacarpophalangeal crease to the proximal phalanx can provide a skin lengthening of about 15 mm. The incision can be extended to the palm in order to remove the other fascial lesions. It was used mainly on the little finger (9 times out of 10). A single flap was usually made, but one may use two flaps from contiguous fingers, and even raise two flaps from the same finger. A comparative study with "Z" plasties was carried out. Out of 131 "Z" plasties with an average lack of extension of 126 degrees, the rate of improvement was 57%. The rotation flap was used in 141 cases with an average lack of extension of 140 degrees, and the rate of improvement was 79%. This flap seems to be an interesting procedure in the surgical treatment of severe Dupuytren contractures and seems more efficient than "Z" plasties. PMID- 9336612 TI - Pathogenesis of Volkmann's ischemic contracture of the first web space and treatment. AB - Reporting three cases of sequelae of Volkmann's ischemia of the first web space of the hand, following barbiturate overdose, the authors suggest several pathogenic hypothesis. The major factor is obviously an elective compression of the top of the first commissure on its dorsal aspect but it due to a compartmental syndrome or to a stoppage in the blood flow? The second explanation seems the best; the radial artery compressed at the top of the first commissure over the first dorsal intermetacarpal ligament is not able to vascularize the dorsal digital artery of the thumb and the first dorsal interosseous artery. These arteries being terminal in type, an elective muscle necrosis is takes place in the adductor pollicis and the first dorsal interosseous. This phenomenon being increased in this last muscle by a simultaneous compartmental syndrome. A precocious diagnosis must be made during the acute stage and some logical surgical procedures performed in emergency: fasciectomy, muscle excision and fixation of the trapezometacarpal joint in opposition with a pin. PMID- 9336613 TI - Acute septic arthritis of the fingers. A clinical study of 87 cases. AB - The authors analyse a clinical series of 87 operated cases of acute septic arthritis of the fingers. Arthritis followed either a direct neglected punctiform inoculation or an adjacent infection (subcutaneous felon), or a serious open wound having received emergency surgery. During surgery a careful assessment was made of the extent of the infection, indicating the condition of the cartilage and that of the articular and periarticular soft tissues. Treatment was either conservative (surgical drainage, synovectomy and closure) or non-conservative (arthrodesis, articular resection or even amputation). The short term results were evaluated on recovery from infection and the long term results (after six months) on the joint function. There is a correlation between the causative factor, the pathology and the prognosis: direct punctiform inoculations operated upon at an early stage and simple joint wounds did not have a regional infection; in all other cases there was one. Conservative treatment was limited to those patients with no regional infection. In these cases, a localized abnormal aspect of the cartilage did not prove to be irreversible. Thus the prognosis was good only in cases of direct punctiform inoculations operated upon at an early stage. In the other cases, either the conservative treatment failed, or a non conservative treatment was decided upon immediately. Severe joint infection leads to residual lack of motion, and the destruction of the articular surfaces may induce the surgeon to perform an arthrodesis. In the early stages of arthritis, surgical attempts have been made at controlling the infection and preserving articular motion (1, 2). The few works published on that matter deal with larger limb joints. In this paper we present a series of 87 cases of acute septic arthritis of the fingers. PMID- 9336615 TI - A radiological study of the wrist centers of rotation. AB - To ensure the total replacement of wrist, it seems necessary to determine the main kinematic specifications of mobility and stability of this joint. After a review of the literature, the authors have determined, by the classical X-rays graphic method, the positions of both radial-ulnar deviation and flexion extension centers, on 26 alive normal wrists. The data, in some ways different from Youm's, introduce to the conditions of stability of any wrist in a functional situation. The center locus (on the forearm axis in a front view, quite anterior in a lateral view) is, in some ways, an answer to the question of muscular equilibrium during prehensile efforts, so to the amount of strains the joint must bear, and consequently to the design of a total wrist prosthesis. PMID- 9336614 TI - Fractures of the fifth metacarpal neck. AB - The authors reviewed 129 cases of fractures of the neck of the fifth metacarpal. The radiological work-up of this fractures should include an anterio-posterior film with the hand supinated, and an oblique ulnar view with the hand pronated. This fractures can be separated into 4 categories: true cervical fractures; cervico-cephalic fractures; cervico-diaphysary fractures; and epiphyseal separations. In 78 patients an immediate mobilisation was performed. 51 fractures have been treated by multiple medullary pinning because of an unacceptable volar angulation associated with mal-rotation. Good and excellent results were obtained in 86% of cases. PMID- 9336616 TI - Round table on Kienbock's disease. PMID- 9336617 TI - [Arterial vascularization of the semilunar bone]. AB - The gross examination of 41 dried lunates and the dissection of 50 minimum injected forearms allowed the author to give a general statement of the blood supply of the lunate. On the volar aspect of the hand, 2 or 3 vessels coming from the ramus carpeus volaris and/or the radial artery run downward and penetrate the bone through a big foramen and several smaller around. On the dorsal aspect, 2 or 3 minute branches arise from the dorsal carpal arch and penetrate the triangular posterior surface of the lunate beneath the carpal joint; more infrequently 2 twigs may be traced along both scaphoid-lunate and triquetrumlunate joints. The interosseous artery sends some conspicuous branches to the posterior margin of the radius and the dorsum of the lunate. The volar group appeared to be the most important contributor to the blood supply and the big volar foramen should be named "hilus" of the lunate. It must be emphasized that the nutrient vessels enter the foramen above the upper margin of the transverse carpal ligament; so, no crowding in the carpal tunnel can be liable for "avascular necrosis" of the lunate. PMID- 9336619 TI - Longitudinal pinning of fractures of the base of the first metacarpal. AB - From an experience of 120 fractures of the base of the first metacarpal bone, the authors evaluated the functional results of in 40 patients followed-up. Most of these patients had been treated by Bunden's method of longitudinal pinning. If one eliminates the comminuted fractures and the Bennett's fractures with a single large fragment treated by screw fixation the method of longitudinal pinning restores normal function to the first interdigitalspace Furthermore, the development of secondary arthrosis of the trapezo-metacarpal joint is rare. PMID- 9336618 TI - Screw or mini-plate fixation in fractures of the first metacarpal. Experience with thirty-nine cases. AB - Re-establishing normal osteoarticular anatomy, and early mobilization are the principal elements in the successful treatment of fractures of the base of the first metacarpal. To achieve these goals in thirty-nine fractures rigid fixation was obtained by screw alone, or by a mini-plate. Seven Bennett's fractures have been treated by screw fixation when the size of the fragment has made it possible. Extra-articular fractures have been treated, either by screw alone, in oblique fractures (7 cases). Or by mini-plate, T or L shaped, in transverse fractures (23 cases). The use of these techniques have been extended, to comminuted fractures but is not used in Bennett's fractures when the fragment is too small to allow the screw to be anchored in it without its being shattered. PMID- 9336620 TI - First results of arthroplasty of the wrist by Swanson's implant. Twenty-five cases. AB - We have carried out 25 arthroplasties of the wrist using the Swanson implant since 1976. The indications were primarily in rheumatoid arthritis. The operation was done for severe pain and deformity of the wrist. Results were analysed in 19 patients with a follow-up of 21-54 months. Post-operative pain relief is good. Although mobility is fair, the return of the balance of wrist motions has been recovered. The results of the Swanson implant arthroplasty are compared to dorsal synovectomy and wrist arthrodesis. PMID- 9336621 TI - Treatment of recent lesions of the dorsoradial compartment of the metacarpophalangeal (MP) joint of the thumb. AB - Lesions of the dorsoradial compartment of the metacarpophalangeal joint of the thumb, are encountered more rarely than rupture of the internal collateral ligament and are considered a less serious or complex lesion. However, the alteration of the various structures, at the level of the metacarpophalangeal joint, produces, a severe deformity. It is an injury that requires early surgical repair of these structures. The author describes the physiopathology of the injury and the surgical technique for correcting the deformity on the basis of a series of operated cases. PMID- 9336622 TI - Ulnar neuritis in Hansen's disease results of fifty neurolyses in the arm and elbow. AB - Fifty cases of ulnar nerve neuritis in Hansen disease are reported. The authors analyse the type of lesion, the clinical feature, the treatment, and the results of neurolysis. Many points are emphasized: the requirement to an association antileprosy, chemotherapy and corticotherapy with a careful neurolysis; pain and paresthesia were relieved immediately after neurolysis, recovery within two years after neurolysis; the amount of recovery was directly related to the extent and stage of involvement of the nerve; and thus the sooner procedure give the best result. PMID- 9336623 TI - Current aspects of tuberculous tenosynovitis. A report of six cases. AB - Tuberculosis of tendon sheaths of the hand was rare before the advent of antituberculous drugs. It seems now-a-days exceptional, for the published series become more an more uncommon and smaller. Therefore, the diagnosis is often an operative surprise on a patient with a painless, gradually developing tenosynovitis. No patient suffered from other active tuberculosis. But two of them had an old history of tuberculosis (knee and kidney). Analysis of other cases is inconclusive about the relation ship between trauma and tuberculosis. Histology and cultures establish the diagnosis. Surgical treatment consists in large excision of involved tissues. The damages provoked by the illness are often important, with sometimes tendon ruptures. However, functional results are surprisingly not so bad. Antituberculous drugs are the backbone of the surgical treatment for several months. PMID- 9336624 TI - [Arthrolysis of the metacarpophalangeal joints in post-traumatic stiffness of fingers in extension]. AB - In cases of post-traumatic stiffness of the metacarpo-phalangeal joints in extension, it is possible to safely perform an arthrolysis with or without tenolysis while preserving the continuity of the extensor mechanism. This paper describes the surgical approach and technique of arthrolysis of the metacarpo phalangeal joints. Proper operative management of post-traumatic stiffness of the long fingers in extension can improve the results in these cases. PMID- 9336625 TI - Radiolunate arthrodesis. Factor of stability for the rheumatoid wrist. AB - Spontaneous radiolunate arthrodesis found in nearly 13% of rheumatoid wrists confer on this joint a durable physiologic orientation with reduced but sufficient mobility. Twelve cases of this type of fusion were studied radiologically and clinically with an average follow-up of 5 years. The authors advise surgical arthrodesis as a supplementary procedure with synovectomy every time there is instability of the carpal articulation with ulnar disalignment. Seven cases are reported, 4 of which are presented in detail. The operative technic and indications are discussed briefly. PMID- 9336626 TI - Arthroplasty of the rheumatoid wrist by silicone implants. Experience with forty cases. AB - The authors report their first evaluation of 40 arthroplasties of the wrist with the Swanson implant. The indication for this procedure was principally severe damage to the wrist joint. The radiologic findings have been summarized. In regard to operative technic, there have been no major problems. In 20 cases also arthroplasty of the head of the ulnar was done. There were no significant early complications. The clinical results after 14 months, with a range of 6 to 45 months, was always satisfactory in regard to pain and the preservation useful flexion and extension. So far, tolerance to the implant seems to be satisfactory. The joint narrowing, regularily found in the new articulation, does not detract from the long-term results. Thus the authors retain a favorable impression of this arthroplasty which should find a place in the treatment of badly damaged rheumatoid wrists. In addition to relieving the pain, it preserves mobility that would be sacrificed with an arthrodesis. Among the associated procedures, the authors have been disappointed with arthroplasty of the head of the ulna and have given it up. PMID- 9336627 TI - Isolated traumatic dorsal dislocation of the distal radio-ulnar joint. Report of seven cases. AB - The authors report seven cases of isolated dorsal dislocations of the distal radio-ulnar joint. Four were recent dislocations and three were old, chronic dislocations. Acute dislocations were treated surgically. An anatomic study on human cadavers enabled us to define the injuries responsible for radio-ulnar dislocations. An electromyographic study of the extensor carpi ulnaris confirmed its role as a dynamic stabilizer of the ulnar head. Two of the three old dislocations were revised after an attempt at ligamentous reconstruction. These patients benefitted from a resection of the ulnar head. The third patient one underwent a Sauve-Kapandji procedure. Cadaver studies underline the fact that ligamentous reconstruction procedures are actually tenodeses. PMID- 9336628 TI - Isolated compression of the motor branch of the ulnar nerve. AB - This paper reports four cases of isolated compression of the deep branch of the ulnar nerve, and reviews the various etiologies (tumor, malformation and microtraumatism). The authors stress the importance of the electric examination, particularly conduction velocity in the palm in localizing the nerve lesion. Surgical exploration in these cases revealed a Giant cell tumor, two synovial cysts, and an abnormal fibrous band stretching between the flexor digiti quinti brevis and the opponents digiti quinti. Postoperative courses were uncomplicated and every patient recuperated quickly and completely. Early surgical exploration in such cases is mandatory. PMID- 9336629 TI - The metacarpo-phalangeal arthrodesis of the thumb according to the tension-band principle: indications and technique. AB - Arthrodesis of the thumb MP I remains a good procedure with numerous and various indications. It is especially valuable in heavy workers. The osteosynthesis, according to the tension-band principle, is stable and enables normal function of the hand after wound healing. The return to heavy work, however, requires at least 4 weeks and a pain free thumb. PMID- 9336630 TI - Thirty cases of duplication of the thumb. Operative results. AB - Thirty cases of duplication of the thumb have been seen during a period of 8 years. Twenty-four children underwent surgery for a total of 25 operations, and these are reviewed after a follow-up of at least a year. Wassel's type IV was the most frequent type, occurring in 14 cases. There were 23 primary operations, and 2 reoperations for frontal deviation, which were secondary to operations done elsewhere. The average age at the time of operation was 16 months, the mean follow-up is 4 years (1 to 8 years). The results are analyzed according to Wassel's criteria. Regardless of the location of the duplication, function is unaffected, the structure of the thumb is good, and the parents are satisfied. However, examination revealed some loss of mobility in 10 cases, shortening of the thumb in 5 cases, instability of the collateral ligament in 3 cases and axis deviation in 10 cases. Six of these were clinodactyly at the IP, 1 a clinodactyly at the MP and 3 Z-shaped deformities at the IP and MP. The two reoperations were done to correct a significant loss of alignment, and in each the result was upgraded from "poor" to "fair". It seems that the end results are determined at the original operation which should be done before the 18th month. The thumb with the least function, usually on the radial side, is resected and the remaining component reconstructured. This includes centralization of the insertions of the extrinsic muscles, and reinsertion of the thenar muscles. Immobilization by pinning is usually done due to the need for an osteotomy during the surgery for realignment of the axis of the thumb. It must, however, always be remembered that this surgery is cosmetic rather than functional. PMID- 9336631 TI - Nerve anastomoses with human fibrin. Preliminary clinical report (56 cases). AB - Since 1980, 56 peripheral nerve repairs have been done with fibrin. For technical reasons, combined anastomoses have been chosen in brachial plexus repairs (23 cases), fibrin alone being used in most other cases (8 free flaps, 17 main trunks, 8 digital nerves). As a whole, results compare evenly with the so-called classical repair methods using stitches. The adhesive method's main advantage is the gain in operative time, without impairing precision. Secondary benefits, such as hemostasis and easier stabilization of small grafts, can be achieved. One question remains: what becomes of fibrin? The survey of present cases would tend to prove that axonal growth through the second anastomosis is impeded proportionally to the length of the graft. The possible action of fibrin in the alteration process leading to a sclerotic diaphragm is not elucidated to this day. Experimental as well as clinical research must be carried on, in order to improve this new way of repairing nerves. PMID- 9336632 TI - Late ulnar paralysis. Study of seventeen cases. AB - Seventeen cases of late ulnar paralysis treated by neurolysis-transposition are reported. The clinical characteristics of these paralysis are emphasized. A very prolonged symptom free interval, a rapid onset and a severe involvement. The ulnar transposition was most often done subcutaneously. Cubitus valgus and definite nerve compression proximal to the arcade of the flexor carpi ulnaris muscle are almost always present. The results as regards the neuropathy are notable: no patient is completely cured and only half are improved. An anatomical study of the nerve path shows the essential role, in the compression of the nerve, of the muscular arcade of the flexor carpi ulnaris muscle which acts in a way similar to the bridge of a violin. Hence, opening it longitudinally is the principal procedure of the neurolysis. This should be routine before the first signs of neuropathy occur in an elbow whose axis is out of alignment as a sequela of a childhood injury. PMID- 9336633 TI - Long-term results in the treatment of fracture-dislocations of Galeazzi in adults. Report on twenty-nine cases. AB - The authors report 29 cases of a true Galeazzi fracture, (i.e. displaced fracture of the radial shaft and disruption of the distal radioulnar joint). In 1/4 of the cases, dislocation was overlooked and the injury was mistaken for a so-called "isolated" fracture of the radius. By accurate open reduction and compression plating of the fracture, both the torn radioulnar ligaments and the articular disc could be repaired and healed. Additional percutaneous Kirschner pinning across the ulna and the radius in order to avoid redislocation, does not seem to be necessary. It is important, however, to hold the reduction of the radioulnar dislocation in a plaster cast for 4-6 weeks, since the 8 persistent displacements of the ulnar head always resulted in a lack of pronosupination of more than 25 degrees. In these cases, pain and disability may require later surgical management. Late resection of the ulnar head or a Sauve-Kapandji procedure which yield an obvious cosmetic and functional improvement, are preferred to any immediate surgical repair of the radioulnar ligaments. This operation was carried out 3 times, but failed twice. Nevertheless out of 25 patients reviewed after a mean follow up time of 6.5 years, the results were gratifying in 20 who could resume their previous occupation 4 to 12 months postoperatively. PMID- 9336634 TI - Occupational "cramps" of the upper limb. AB - Problems involving the coordination of movements of the hand analogous to the "writers cramp" have been seen in musicians and other professions. They occur in individuals with muscular imbalance, who assume abnormal body postures affecting not only the involved limb but also the spine and pelvis. These patients also often have a particular psychological make up. Treatment is based on the re training of their muscular actions and on teaching the patients to avoid assuming abnormal stances or postures. PMID- 9336635 TI - Fixed post-traumatic flexion-contractures of digits. Review of thirty-three cases. AB - A fixed post-traumatic flexion contracture of a finger is usually secondary to multiple previous operations. We have observed that a former flexor tendon laceration is not constant and is missing in 18% of our cases. The flexor tendons are, nevertheless, always involved in the contracture. A volar skin contracture was present in all cases, but only in half of them was noted a retraction of the volar components of the PIP joint. This articular involvement has no statistical correlation with the time elapsed from the onset of the contracture. We have reviewed 33 cases of post-traumatic flexions contractures of the digits all secondary to volar trauma. In every case there was at least a flexor tendon adhesion and skin contracture. They have all been submitted to both objective and statistical analysis. Results have been evaluated by comparison between the normal functional range of motion for each digit and the actual post-operative active range of motion. On the basis of our study we conclude that the age of the patient is an important prognostic factor. We obtained 75% satisfactory results in patients younger than 27 years, but only 22% in the older group. Good results are more easily obtained in radial (65%) than ulnar digits (31%). While the authors rated 39% of the results bad, half of the patients in this group were satisfied with the result. A volar PIP joint release has been necessary in half of the cases with no significant secondary joint stiffness. A skin flap is necessary to cover the cutaneous defect secondary to the release. There is no statistically significant advantage to cross finger flaps. Therefore we feel that local flaps are indicated except in the cases where local scar tissues would not make it, feasible. The prognosis is independent of the number of previous operations and of associated nerve lesions. Therefore amputation is not the only solution for a multi-operated finger fixed in flexion. PMID- 9336636 TI - Krukenberg's operation. Indications and limitations. AB - The Krukenberg operation--creating a sensory forceps--is indicated principally in bilateral below elbow amputation with blindness. Other indications are proposed, in particular in children. The limitations to this procedure are essentially aesthetic problems and should not be minimized. The operative technique is described in details. PMID- 9336637 TI - Long-term results from metacarpophalangeal arthroplasty. AB - The authors have been using Swanson's prostheses for MP joint replacement in rheumatoid arthritis since 1968. After thirteen years of experience, they present an analysis of these prostheses from a clinical and radiological point of view. 88 joints replacements in 20 patients are reviewed. These operations were performed between 1968 and 1976. The average follow-up is nine years. The authors compare their results with those obtained in a previous study made in 1975. They show that a replacement arthroplasty with a Swanson type prosthesis, imparts considerable benefits to the patient in the form of complete disappearance of pain, improvement in function, and a more normal looking hand, in spite of radiologic deterioration. PMID- 9336639 TI - Restoration of thumb-index pinch by a double tendon transfer following ulnar nerve paralysis. AB - Repair of ulnar nerve lesions by suture or grafts often gives good results, but in some cases there persists a long-term isolated paralysis of the muscles of the first web space. There is certain to be a resulting loss of function from loss of the thumb-index pinch which can be corrected by tendon transfers. In 11 cases the authors have used two tendon transfers: the short extensor of the thumb to re establish the function of the first dorsal interosseous muscle, and the extensor indicis proprius to improve adduction. In every case there was notable improvement in the function of the hand. PMID- 9336638 TI - The forearm fascia flap. AB - A new subcutaneous flap technic, derived from the chinese forearm flap has been demonstrated. It has been used to cover large cutaneous defects of the hand, especially when the extensor tendons were exposed. An excellent functional repair has been obtained in a single procedure avoiding large scar formation usually seen on the forearm after classical chinese fasciocutaneous flap. PMID- 9336640 TI - Systemization of the vascularization of the ulnar nerve in its upper arm. AB - Vascularized free nerve grafts make possible the repair of extensive defects of large nerve trunks. Since the original observations of Taylor and Ham in 1976 many cases have been published. The ulnar nerve in the upper arm in most cases has a simple arteriovenous pedicle the anatomy of which has been precisely defined by cadaver dissections and intravascular injections. The arterial supply, 47 times out of 50, is the proximal ulnar collateral and 2 times the distal collateral ulnar artery. It takes its origin from the medial side of the brachial artery in the upper or middle third of the arm. Its external diameter is on the average 1.8 mm at its origin. The accompanying vein enters a brachial vein 2 to 3 cm below the origin of the artery. The removal of the graft is done through a straight incision on the inner aspect of the arm. The brachial artery is dissected from above downward and its medial branches noted. The nerve and its arteriovenous pedicle are separated in a block along with adjacent cellular tissue by dissection from below upwards. The average length of the pedicle thus produced is 13 cm, but a much longer section of the nerve can certainly be taken. A case report illustrates the procedure. PMID- 9336641 TI - The importance of the posteromedial fragment and its specific pinning in fractures of the distal radius. AB - In fractures of the lower end of the radius, the presence of a posteromedial fragment should be recognized if present. This fragment with two articular surfaces, if not reduced, will damage the radiocarpal and inferior radioulnar articulations. The diagnosis needs to be made on 3/4 X-ray views, as the fracture is often mistaken for a Colles' fracture. The author advises reduction and fixation by a method of double pinning. By comparing two sets of statistics, the overall results are shown to be improved without more complications. PMID- 9336642 TI - Surgical treatment of ganglions of the wrist by partial excision of the joint capsule. Report on 303 cases. AB - Histologic examination confirms the nonsynovial nature of cysts of the wrist, generally located on the dorsal surface. There is found a mucoid degeneration of the capsule and surrounding tissue with numerous neighboring microcysts. This is why it is irrational and usually ineffective to treat these cysts by crushing, aspiration or simple excision. These three methods result in frequent recurrences. One hundred and eighty cases were operated by excision of the cyst and all the neighboring involved tissue including in particular a disk 2 to 4 cm in diameter from the capsule of the joint. Almost all were cured, except for 3 recurrences, and any stiffness or weakness found was present preoperatively. A very detailed radiologic study (an average of 9 films per patient) made 3 years or more after operation showed there was no resultant carpal instability. In spite of its radial character this is the operation of choice in view of its efficacy and in cases where treatment is justified because of appearance or interference with function. PMID- 9336643 TI - The carpal tunnel syndrome in chronic dialysis patients, it is a late complication of the arterio-venous fistula? AB - Out of 100 patients undergoing chronic hemodialysis in Lausanne (Switzerland), 12 developed a carpal tunnel syndrome (i.e. 8 men and 4 women, from 34 to 76 years old). Out of 66 patients with an arteriovenous fistula for less than 4 years, it is interesting to note that only 3 carpal tunnel syndrome were observed; whereas, out of 34 hemodialysis patients being dialyzed more than 4 years, 9 of them showed that syndrome. The symptomatology of the carpal tunnel syndrome is described by the authors. It has always been confirmed by EMG. On 10 patients, the symptoms were so acute that they needed a decompression of the median nerve. Five cases were bilateral. The operation is performed under axillary block, without tourniquet. The results were very satisfactory. Paresthesias disappeared after a few hours or a few days following the operation. No relationship could be established between CTS and the type of nephropathy, severity of polyneuropathy, Ca and P metabolism, vascular access complications, efficacity of dialysis, fluid overload or medical treatment. The authors are investigating the etiology of the carpal tunnel syndrome, ship hypothesize particularly concerning the direct or remote relation between the carpal tunnel syndrome and the arteriovenous fistula. PMID- 9336644 TI - Osteosynthesis of fractures of the base of the first metacarpal by an external fixator. AB - Among the various methods for treating fractures of the base of the first metacarpal, use of the external fixator appears to be of interest for it maintains a satisfactory reduction while respecting the arch of the first web interspace as well as allowing early mobilization of all joints not involved in the injury. The miniaturization of the material, today possible, facilitates its use. The experience with 20 cases gained at the CTO de la CRAM de Strasbourg is presented. The authors discuss the utilization of the method, the practical means of doing it and their results. Its limitations are indicated as well as its special indications in comminuted and compound fractures. PMID- 9336645 TI - Proposal of a method for objective evaluation of results following flexor tendon repair. AB - The author is critical of the usual methods of evaluating the results after suturing of the flexor tendons. The methods in general based on active articular mobility of the finger, do not take into account the condition of the hand examined, and are not precise on taking the measurements. He proposes a method which is simple and more objective for it is based on the measurement of the total function of the finger compared to that of the same finger on the other hand. The results, expressed in percents, are classified into three groups (A, B, C) whose limits vary whether the lesion is an isolated one of the deep flexor tendon or of the both flexors. PMID- 9336646 TI - Early mobilization of fractures of the metacarpals and phalanges. AB - Early mobilization of fractures of the metacarpals and phalanges is advocated. It should begin as soon as the stability of the fracture and the condition of the soft tissues permits. It has to be adapted to each case, and often consists of mobilization of the distal phalanges with the wrist and metacarpophalangeal articulations immobilized in a protective position. It is effective in preventing stiffness. Because it is a specific technique, it must be entrusted to specially trained physiotherapists, and requires regular and repeated clinical and radiologic follow up by the surgeon. PMID- 9336647 TI - Functional brace in the treatment of diaphyseal fractures of the proximal phalanges of the last four fingers. AB - The authors describe a system of functional brace for the treatment of displaced fractures of the proximal phalanx of the fingers. Using four digit syndactylization this technique allows simultaneous stabilisation of the fracture and early active mobilization of the proximal interphalangeal joint, where stiffness is known as the principal impairment after such fractures. Among 14 cases treated by this method, fracture healing has been obtained in all cases with only one significant malunion. No severe stiffness has been observed and active motion at the PIP joint has been in a useful sector of 25 degrees and 75 degrees at least in all cases. PMID- 9336648 TI - Epithelioid sarcomas. Three cases. AB - The authors present two cases of epithelioid sarcoma of the hand and one of the foot with clinical interest. This recently described tumor (Enzinger, 1970), is relatively rare. It occurs in the hand, forearm, pretibial region and foot and affects principally young adults. It should be emphasized, and this is borne out in the literature, that this tumor may appear perfectly benign and often has a course of long duration. The difficult of clinical and especially of histologic diagnosis as well as the difficulty of determining the boundaries of extension of the tumor makes it necessary to carry out radical surgery (amputation or rarely block excision). Every author agrees that local excision is to be condemned, there being an 85% recurrence rate. Spread of the tumor is by way of the fascial planes and tendon sheaths. Lymphatic and pulmonary metastases occur particularly when there is vascular invasion. PMID- 9336649 TI - Epithelioid sarcoma. AB - Epithelioid sarcoma is a malignant soft tissue tumor characterized by its propensity to occur in the distal extremities as a nodular lesion and its slow and asymptomatic growth. Microscopically, the diagnosis is difficult. Therefore the interval between onset of symptoms and diagnosis averages one to three years. 10 years survival after onset are not uncommon whatever the treatment chosen. Having to treat such a tumor of a hand, 8 years after onset, we decided a local radical excision by micro-surgery instead of amputation or mutilating excision. Long standing clinical follow-up will add valuable information as to the cure of the upper extremity. PMID- 9336650 TI - Rupture of the flexor tendons of the little finger in fractures of the hook of the hamate bone. Report of two cases. AB - Two cases of rupture of the flexor tendons of the little finger associated with a fracture of the hook of the hamate bone are presented. The preoperative diagnosis had not been made. Excision of the fractured hook and repair of the tendons by a short graft from the palmaris longus in one case and by the transfer of the superficial flexor of the ring finger in the other case gave a good final result. The authors remark on the rarity of reports of this fracture in the literature, the frequent occurrence of nonunion and the two complications which are exceptional; rupture of the flexor tendons and compression of the deep branch of the ulnar nerve. The fracture is mainly encountered in sports that require the grasping of a handle (tennis, golf, hockey, squash). The diagnosis is often missed at the onset because of not using the correct X-ray positioning: special incidence for the carpal tunnel view and a 3/4 view with the wrist in 45 degrees of supination and forced radial deviation. PMID- 9336651 TI - Simple arthrolysis for flexor rigidity of the proximal interphalangeal joint. AB - The authors report 19 cases of simple arthrolysis of the proximal interphalangeal joint for flexion rigidity. Their cases were limited to isolated lesions of the joint without any flexor or extensor tendon involvement. The technique is described, and the importance of postoperative physical therapy is stressed. The results in these cases, in contrast with those in complicated cases of rigidity, are very satisfactory. The etiology is primarily sprains and dislocations of the proximal interphalangeal articulation, immobilized for too long a time in flexion (in the so-called "functional position"). PMID- 9336652 TI - Skin problems in the treatment of the finger "en crochet". AB - The study of 33 fixed flexion contractures of the finger shows that after tenolysis or teno-arthrolysis a skin gap appears when the finger is extended. Only 10 cases out of 33 had previous cutaneous involvement. The necessary skin plasties have been 14 heterodigital flaps and 19 local dorsal-lateral flaps. Results are comparable in both series and the surface of the skin defect has no influence on the choice of the type of flap. Therefore, whenever possible, a local flap is preferred to a distal flap. PMID- 9336653 TI - Two-step reconstruction of the flexor tendons (Hunter's technique) in the treatment of fingers "en crochet". AB - The authors used Hunter's technique to treat 16 cases of fixed-flexion deformity of the finger (with irreducible flexion of the joint greater then 70 degrees), secondary to lesions of the flexor tendons. The flexion deformity was corrected in 11 of the 16 cases, and active movement greater than 70 degrees was obtained in only 7 cases. Technical difficulties (particularly cutaneous problems), frequent complications, prolonged reeducation are the reasons that the indications for its use are exceptional, and why there is need for excellent cooperation by the patient. PMID- 9336654 TI - Total anterior teno-arthrolysis. Report of 72 cases. AB - This report describes a new technique for treatment of the chronically flexed fingers which applies in particular to fingers previously operated on several times and now presenting cutaneous, tendinous and joint problems. It consists in releasing the entire flexor apparatus through a full length lateral digital incision and a sub periosteal dissection. The volar plates of PIP and DIP are released as a whole with the flexor apparatus. The extended finger and flexor apparatus then are allowed to heal in a new relationship. Straightening of the finger is always possible. The range of motion is maintained or increased. This technique can also be used in stiff PIP joints and in certain serious forms of Dupuytren's contracture. 56 cases are reviewed with 78% with good or fair results. PMID- 9336655 TI - Biologic characteristics of breast cancer detected by mammography and by palpation in a screening program: a pilot study. AB - OBJECTIVE: To compare the histopathologic features and expression of p53 and c erb B2 in the tumours detected by mammography only (clinically occult tumours) and the tumours detected by a nurse examiner (clinically palpable tumours). SETTING: London branch of the Ontario Breast Screening Program, which uses both clinical breast examination and mammography as screening methods. INTERVENTIONS: Pathologic review and immunohistochemical staining of all tumours detected between 1990 and 1993. OUTCOME MEASURES: Categorization of tumours by detection method and analysis of tumour size, grade, type, lymph node status and c-erb B2 and p53 expression in each group. RESULTS: From 1990 to 1993, 131 tumours were detected in patients ranging in age from 50 to 85 years (median 63 years). Sixty seven occult tumours and 64 palpable lesions were detected. The occult tumours were significantly smaller (1.34 cm v. 2.29 cm, p < 0.0001) than the palpable ones and included a higher proportion of special-type lesions and ductal carcinoma in situ (43.3% v. 10.9%, p < 0.0001). Occult invasive carcinomas were of lower grade than palpable carcinomas (68.4% grade 1, 21.1% grade 2, 10.5% grade 3 v. 32.8% grade 1, 36.1% grade 2, 31.1% grade 3, p < 0.0001). Fewer occult lesions showed axillary nodal metastases (19.6% v. 40.6%, p = 0.02). No statistically significant differences were found for p53 or c-erb B2 positivity between the 2 groups. CONCLUSION: Tumours detected by different screening methods in a screening program have different pathologic characteristics. PMID- 9336656 TI - High frequency of neoplasia in patients with autoantibodies to centromere protein CENP-F. AB - OBJECTIVE: To study the clinical features of patients with autoantibodies to centromere protein CENP-F and the frequency of CENP-F autoantibodies in patients with various diseases. DESIGN: Retrospective clinical and serologic study. METHODS: Thirty-six patients with anti-CENP-F were identified by a characteristic pattern of indirect immunofluorescence (IIF) on HEp-2 cells. Fifty patients with melanoma, 50 with breast cancer, 10 with lung cancer, 354 with systemic sclerosis, 120 with systemic lupus erythematosus and 50 with rheumatoid arthritis were also studied. Recombinant proteins were produced from 5 CENP-F cDNA clones representing amino acids 2192-3317 (p-F1), 5561-7126 (p-F2), 5892-6883 (p-F3), 7538-10,116 (p-F4) and 9242-10,096 (p-F5). The presence of CENP-F antigen was studied in a breast carcinoma cell line, cryosections of breast carcinoma, normal breast tissue and tonsils. RESULTS: Twenty-two of 36 patients with CENP-F antibodies had neoplasms; breast (9/22) and lung (5/22) cancer were the most common diagnoses. Thirty-three sera were available for further study; when tested for reactivity to the recombinant peptides, the sera of 21 of 21 patients with neoplasms and 5 of 12 patients with other diseases bound the C-terminal p-F4 peptide. When the terminal third of the p-F4 peptide (p-F5) was studied, a significant difference in pattern of reactivity was not detected. By comparison, the frequency of reactivity with peptides representing other domains of CENP-F was less than that with p-F4 (p-F2 > p-F3 > p-F1). CENP-F autoantibodies were not found in any of the control sera from patients with systemic lupus erythematosus, rheumatoid arthritis or systemic sclerosis or in unselected sera from various malignancies. CENP-F antigens were identified in breast carcinoma tissue but were rarely observed in normal tissues. CONCLUSIONS: A high proportion of individuals with CENP-F antibodies have neoplasia, and there is a bias among their sera for reactivity with determinants in the carboxy terminal domain of CENP-F. CENP-F antigens appear to be highly expressed in malignant tissues. PMID- 9336657 TI - Management of unstable coronary syndromes in patients with previous coronary artery bypass grafts following coronary angiography. AB - OBJECTIVE: To characterize patients who had undergone previous coronary artery bypass grafting (CABG) and who were admitted for coronary angiography for unstable coronary syndromes, to determine the long-term therapy selected for these patients and to assess the outcomes of the intervention. DESIGN: Descriptive retrospective study. SETTING: A university-affiliated tertiary care institution. PATIENTS: A total of 129 patients with 1 previous CABG who underwent coronary angiography for myocardial infarction or unstable angina in 1991. OUTCOME MEASURES: Information regarding initial CABG, indications for cardiac angiography, cardiovascular risk factors, ultimate treatment selected and outcomes at 1 year were abstracted from patients' charts, and outcomes at 1 year were also determined by a patient survey. RESULTS: Seventy-six patients (59%) were given drug therapy, 28 patients (22%) were treated with angioplasty, and 25 (19%) underwent repeat surgery. During their index admissions, of patients given drug therapy, 4 (5.3%) died from myocardial infarction (MI) and 42 (55%) were discharged without complications; of those undergoing angioplasty, all except 2 were treated successfully (major procedural complications included nonfatal MI in 1 patient [4%] and nonfatal ventricular arrhythmia in 1 patient [4%], as well, reocclusion of the lesions occurred before discharge in 2 patients [7%]); of those undergoing repeat surgery, almost all patients (96%) were discharged, except 1 who died from MI during the postoperative period (there were no procedural complications, but early complications included nonfatal MI in 2 patients [8%], angina in 2 [8%] and nonfatal arrhythmias in 11 [44%]). Eighty seven patients (67%) were available for follow-up at 1 year. Of the patients given drug therapy, 3 (6.4%) had died, 14 (30%) had recurrent anginal episodes and 5 (11%) required either angioplasty or CABG. Of the patients who initially received angioplasty, 15 (63%) had recurrent angina but none died, 12 (50%) underwent repeat angioplasty and 2 (8.3%) required repeat CABG. No patients who received repeat surgery died or required further surgery or angioplasty. Three of these patients (19%) had recurrent angina within the first year. Patients in this category also enjoyed a greater degree of symptomatic improvement of coronary artery disease. CONCLUSIONS: Patients who had a previous CABG and subsequently presented with MI were more likely to be given conservative drug therapy than those who presented with unstable angina. At 1-year follow-up, recurrent angina occurred more often in the patients treated by angioplasty, less often in patients given drug therapy and least in those who underwent repeat bypass grafting. Restenosis remained a problem, and about 50% of patients treated with angioplasty (without intracoronary stenting) required a second angioplasty within the first year. Patients who were candidates for repeat CABG enjoyed greater symptomatic improvement within the first year. PMID- 9336658 TI - Prevention of stroke with perindopril treatment in stroke-prone spontaneously hypertensive rats. AB - OBJECTIVE: To determine the protective effects of perindopril treatment in the prevention of stroke and the relation between preventive effects and the histopathology of the brain and kidneys in male stroke-prone spontaneously hypertensive rats (SHRSP). DESIGN: Prospective animal study. INTERVENTIONS: Beginning at 6 weeks of age, SHRSP were treated with either distilled water (control) or perindopril for different periods (8, 12 or 24 weeks) and at different dosages (1 or 4 mg/kg per day). OUTCOME MEASURES: Regular determination of systolic blood pressure, heart rate and body weight until death; at necropsy, macroscopic and microscopic examinations of the brain and kidneys. RESULTS: Control SHRSP developed severe hypertension (up to 250 mm Hg) by 11 weeks of age and died of stroke within 14 weeks of age. Treatment with perindopril (4 mg/kg per day for 8 or 12 weeks or either 1 or 4 mg/kg per day for 24 weeks) attenuated the blood pressure rise and prevented stroke. In untreated SHRSP, the last blood pressure measurement before the first stroke sign was significantly higher than in SHRSP of the same age treated with perindopril. Withdrawal of the treatment resulted in a rise in blood pressure in all the treatment groups, to approximately 260 mm Hg within 4 weeks. Most of the rats treated for 8 or 12 weeks died within 10 weeks after withdrawal of treatment, whereas those treated for 24 weeks survived up to 43 weeks of age. Treatment also prevented damage to the brain and kidneys and reduced the severity of lesions in the brain and kidneys after treatment withdrawal. CONCLUSION: Treatment of SHRSP with perindopril prevents stroke through the suppression of blood pressure rise and prevention of tissue damage in the brain and the kidneys. Longer treatment decreased the rate of mortality due to stroke after the withdrawal of treatment as well as the severity of lesions in the brain and kidneys. PMID- 9336659 TI - Limiting the Niemann-Pick type C critical region to a 1-cM interval. AB - OBJECTIVE: To refine the position of and isolate the gene responsible for Niemann Pick Type II (NP Type II) disease, an autosomal, recessive neurodegenerative disorder usually affecting children. The underlying biochemical defect results in an impairment in transport of intracellular cholesterol. This disease has been classified into two subtypes, NPC and NPD. NPD and the major complementation group of NPC both map to chromosome 18q11-12; therefore, they are likely allelic variants. The NP Type II gene was previously localized between microsatellite markers D18S44 and D18S1108. DESIGN: Linkage analysis. SETTING: Pathology department of a university-associated hospital. PATIENTS: An NPC family, including proband, parents and sister. OUTCOME MEASURES: NP Type II disease phenotype and biochemical phenotype (cholesterol esterification). RESULTS: DNA from the individuals in the NPC family was genotyped at 12 microsatellite loci from the critical region. The deduced haplotypes identify a meiotic recombinant that has allowed the distal limit of the critical region to be moved from D18S1108 to D18S1101. CONCLUSION: The NP Type II gene lies proximal to the microsatellite marker D18S1101, within the 1-cM interval between D18S1101 and D18S1398. This represents approximately 1.1 mb on the physical map. PMID- 9336660 TI - Effects of insulin on renal function, sympathetic nervous activity and forearm blood flow in normal human subjects. AB - OBJECTIVE: To assess fully the vasodilatory and sodium-retaining effects of insulin. DESIGN: Prospective physiologic study using a dose-response protocol. SETTING: Clinical investigation unit of a tertiary referral hospital. PARTICIPANTS: Six normal, healthy men. INTERVENTIONS: Subjects were given increasing doses of insulin intravenously from 10 to 1200 mU/m2 per minute, using the euglycemic "clamp" technique. OUTCOME MEASURES: Urinary sodium excretion, systemic and renal hemodynamics, plasma norepinephrine levels and forearm blood flow after each dose. RESULTS: Low doses of insulin (up to 20 mU/m2 per minute) produced a significant antinatriuresis (0.18 [SEM 0.05] v. 0.37 mmol per minute at baseline, p < 0.01) and antidiuresis (2.53 [SEM 0.67] v. 6.21 [SEM 1.66] mL per minute, p < 0.01) with no associated changes in renal hemodynamics or sympathetic nervous activity. Subsequent higher doses of insulin improved urinary volume and sodium excretion to above baseline levels associated with renal and forearm vasodilatation, although mean arterial pressure remained unaltered. CONCLUSIONS: Hyperinsulinemia initially causes an antinatriuresis and antidiuresis through a direct action on the renal tubule. The subsequent phenomenon of escape from renal sodium retention may serve as a regulatory mechanism on sodium homeostasis in conditions associated with hyperinsulinemia and sodium retention. PMID- 9336661 TI - Patient-based research in a tertiary pediatric centre: a pilot study of markers of scientific activity and productivity. AB - OBJECTIVE: To characterize patient-based research in a large academic pediatric centre, to examine measures of research activity and productivity among the 44 clinical programs and to examine whether there is a relation among various measures of scientific productivity. DESIGN: Survey. PARTICIPANTS: Clinical programs. OUTCOME MEASURES: Analysis of all patient-based research projects for the 1993-94 and 1994-95 fiscal years, research funding and cumulative citation impact. RESULTS: Only half of the research projects were funded by extramural grants (peer-reviewed or industrial). There were strong and significant correlations among the 3 markers of scientific activity and productivity: funding, peer-reviewed publications and cumulative citation impact. Only small programs with 3 or fewer faculty members with protected time available to develop research programs achieved a citation impact of 30 or more per full-time equivalent position, with larger programs being "diluted" by clinicians performing little or no research. CONCLUSIONS: In the context of patient-based research, quantity of research correlated with measures of quality. This study highlights the need for clinical departments and medical faculties to consider activity and productivity markers in setting standards for patient-based research. PMID- 9336662 TI - Occurrence, distribution, and associations of O and H serogroups, colonization factor antigens, and toxins of enterotoxigenic Escherichia coli. AB - Enterotoxigenic Escherichia coli (ETEC) is a leading cause of infectious diarrhea worldwide. Four categories of antigens have been commonly studied: O serogroup, H serogroup, colonization factor antigens (CFA), and toxins. A database has been complied from published reports of nearly 1,000 ETEC isolates from 18 locations and analyzed to determine the occurrence, distribution, and associations of O serogroup, H serogroup, CFA, and toxin type. Tables listing the associations of antigens are presented. This analysis documents the widespread nature and variety of ETEC. Even the most common combination of antigens, O6:H16 CFA/II LTST, accounted for only 11% of the ETEC isolates in the database. It was isolated from 12 locations. Many phenotypes occurred only once. CFA detection based on enzyme linked antibodies with polyclonal sera is suggested as the preferred assay. A combination of CFA and toxin-based antigens is suggested as the most practical vaccine. PMID- 9336663 TI - Prospects for the development of fungal vaccines. AB - In an era that emphasizes the term "cost-effective," vaccines are the ideal solution to preventing disease at a relatively low cost to society. Much of the previous emphasis has been on childhood scourges such as measles, mumps, rubella, poliomyelitis, and Haemophilus influenzae type b. The concept of vaccines for fungal diseases has had less impact because of the perceived limited problem. However, fungal diseases have become increasingly appreciated as serious medical problems that require recognition and aggressive management. The escalation in the incidence and prevalence of infection has prompted a renewed interest in vaccine development. Herein, I discuss the most recent developments in the search for vaccines to combat fungal infections. Investigators have discovered several inert substances from various fungi that can mediate protection in animal models. The next challenge will be to find the suitable mode of delivery for these immunogens. PMID- 9336664 TI - Uses of inorganic hypochlorite (bleach) in health-care facilities. AB - Hypochlorite has been used as a disinfectant for more than 100 years. It has many of the properties of an ideal disinfectant, including a broad antimicrobial activity, rapid bactericidal action, reasonable persistence in treated potable water, ease of use, solubility in water, relative stability, relative nontoxicity at use concentrations, no poisonous residuals, no color, no staining, and low cost. The active species is undissociated hypochlorous acid (HOCl). Hypochlorites are lethal to most microbes, although viruses and vegetative bacteria are more susceptible than endospore-forming bacteria, fungi, and protozoa. Activity is reduced by the presence of heavy metal ions, a biofilm, organic material, low temperature, low pH, or UV radiation. Clinical uses in health-care facilities include hyperchlorination of potable water to prevent Legionella colonization, chlorination of water distribution systems used in hemodialysis centers, cleaning of environmental surfaces, disinfection of laundry, local use to decontaminate blood spills, disinfection of equipment, decontamination of medical waste prior to disposal, and dental therapy. Despite the increasing availability of other disinfectants, hypochlorites continue to find wide use in hospitals. PMID- 9336666 TI - Progress towards development of a vaccine for amebiasis. AB - The application of molecular biologic techniques over the past decade has seen a tremendous growth in our knowledge of the biology of Entamoeba histolytica, the causative agent of amebic dysentery and amebic liver abscess. This approach has also led to the identification and structural characterization of three amebic antigens, the serine-rich Entamoeba histolytica protein (SREHP), the 170-kDa subunit of the Gal/GalNAc binding lectin, and the 29-kDa cysteine-rich protein, which all show promise as recombinant antigen-based vaccines to prevent amebiasis. In recent studies, an immunogenic dodecapeptide derived from the SREHP molecule has been genetically fused to the B subunit of cholera toxin, to create a recombinant protein capable of inducing both antiamebic and anti-cholera toxin antibodies when administered by the oral route. Continued progress in this area will bring us closer to the goal of a cost-effective oral combination "enteric pathogen" vaccine, capable of inducing protective mucosal immune responses to several clinically important enteric diseases, including amebiasis. PMID- 9336667 TI - A week in the life of a travel clinic. AB - International travel has increased enormously in recent years. With the greater movement of people have come increased encounters with a wide variety of diseases: malaria, dengue, cholera, typhoid fever, Ebola virus, and many more. The need for greater scope, consistency, and knowledgeability in pretravel health care to meet these challenges has been met by the emergence of the discipline of travel medicine. Travelers are well advised to become informed of the risks they face and to take steps to minimize those risks. After reviewing a traveler's medical history and a detailed itinerary, a travel medicine practitioner can offer expert advice on behavioral modifications, immunizations, and chemoprophylaxis regimens which will increase the traveler's margin of safety. The issues most frequently addressed in a travel clinic include treatment of traveler's diarrhea, malaria chemoprophylaxis, and immunizations, for hepatitis A, typhoid fever, tetanus/diphtheria, influenza, pneumococcus, hepatitis B, polio, meningococcus, measles, mumps, rubella, varicella, and rabies. Pretravel consultation must consider the age and underlying health problems of the traveler, the nature of the trip (wilderness, jungle, rural, urban, resort, or cruise), the duration of travel, and the latest available information on the site in terms of disease outbreaks, terrorism, and natural calamities. PMID- 9336668 TI - In search of a selective antiviral chemotherapy. AB - This article describes several approaches to a selective therapy of virus infections: (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU [brivudin]) for the therapy of herpes simplex virus type 1 and varicella-zoster virus infections: (S) 9-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC [cidofovir]) for the therapy of various DNA virus (i.e., herpesvirus, adenovirus, papillomavirus, polyomavirus, and poxvirus) infections; 9-(2-phosphonylmethoxyethyl)adenine (PMEA [adefovir]) for the therapy of retrovirus, hepadnavirus, and herpesvirus infections; (R)-9-(2-phosphonylmethoxypropyl)adenine (PMPA) for the therapy and prophylaxis of retrovirus and hepadnavirus infections; and nonnucleoside reverse transcriptase inhibitors (NNRTIs), such as tetrahydroimidazo[4,5,1-jk][1,4] benzodiazepin-2(IH)-one and -thione (TIBO), 1-[(2-hydroxyethoxy)methyl]-6 (phenylthio)thymine (HEPT), alpha-anilinophenylacetamide (alpha-APA), and 2',5'bis-O-(tert-butyldimethylsilyl)-3'-spiro-5"-(4"-amino-1",2"-oxat hiole- 2",2"-dioxide)pyrimidine (TSAO) derivatives, and thiocarboxanilides for the treatment of human immunodeficiency virus type 1 (HIV-1) infections. For the clinical use of NNRTIs, some guidelines have been elaborated, such as starting treatment with combinations of different compounds at sufficiently high concentrations to effect a pronounced and sustained suppression of the virus. Despite the diversity of the compounds described here and the different viruses at which they are targeted, they have a number of characteristics in common. As they interact with specific viral proteins, the compounds achieve a selective inhibition of the replication of the virus, which, in turn, should be able to develop resistance to the compounds. However, as has been established for the NNRTIs, the problem of viral resistance may be overcome if the compounds are used from the start at sufficiently high doses, which could be reduced if different compounds are combined. For HIV infections, drug treatment regimens should be aimed at reducing the viral load to such an extent that the risk for progression to AIDS will be minimized, if not avoided entirely. This may result in a real "cure" of the disease but not necessarily of the virus infection, and in this sense, HIV disease may be reduced to a dormant infection, reminiscent of the latent herpesvirus infections. Should virus replication resume after a certain time, the armamentarium of effective anti-HIV and anti-herpesvirus compounds now available, if applied at the appropriate dosage regimens, should make the virus return to its dormant state before it has any chance to damage the host. It is unlikely that this strategy would eradicate the virus and thus "cure" the viral infection, but it definitely qualifies as a cure of the disease. PMID- 9336665 TI - Interleukin-12 in infectious diseases. AB - Interleukin-12 (IL-12) is a potent immunoregulatory cytokine that is crucially involved in a wide range of infectious diseases. In several experimental models of bacterial, parasitic, viral, and fungal infection, endogenous IL-12 is required for early control of infection and for generation and perhaps maintenance of acquired protective immunity, directed by T helper type 1 (Th1) cells and mediated by phagocytes. Although the relative roles of IL-12 and gamma interferon in Th1-cell priming may be to a significant extent pathogen dependent, common to most infections is that IL-12 regulates the magnitude of the gamma interferon response at the initiation of infection, thus potentiating natural resistance, favoring Th1-cell development; and inhibiting Th2 responses. Treatment of animals with IL-12, either alone or as a vaccine adjuvant, has been shown to prevent disease by many of the same infectious agents, by stimulating innate resistance or promoting specific reactivity. Although IL-12 may enhance protective memory responses in vaccination or in combination with antimicrobial chemotherapy, it is yet unclear whether exogenous IL-12 can alter established responses in humans. Continued investigation into the possible application of IL 12 therapy to human infections is warranted by the role of the cytokine in inflammation, immunopathology, and autoimmunity. PMID- 9336673 TI - Differential diagnosis between attention-deficit/hyperactivity disorder and pervasive developmental disorder--not otherwise specified. AB - The lack of clarity in diagnostic classification and the lack of specificity of assessment devices deter accurate identification of children with Pervasive Developmental Disorder-Not Otherwise Specified (PDD--NOS). The current study was designed to assess the utility of the Personality Inventory for Children (PIC) and the Conners Parent Rating Scale (CPRS-48) in differentiating PDD--NOS from Attention Deficit Hyperactivity Disorder (ADHD), disorders with overlapping symptom constellations. Subjects were 44 children recruited from a tertiary-care center with the diagnosis of ADHD or PDD--NOS. Results showed significant differences between groups on PIC scales assessing internalizing behaviors, social skills, and unusual affect and behavior. There were no group differences on the externalizing or learning scales of the CPRS-48. Discriminant function analysis using preselected PIC variables yielded a correct group classification of 92.7%. The PIC appears to be a useful tool in the differential diagnosis between PDD--NOS and ADD, while the CPRS-48, a commonly used screening measure for attentional and behavioral disorders, does not. PMID- 9336669 TI - Rickettsioses as paradigms of new or emerging infectious diseases. AB - Rickettsioses are caused by species of Rickettsia, a genus comprising organisms characterized by their strictly intracellular location and their association with arthropods. Rickettsia species are difficult to cultivate in vitro and exhibit strong serological cross-reactions with each other. These technical difficulties long prohibited a detailed study of the rickettsiae, and it is only following the recent introduction of novel laboratory methods that progress in this field has been possible. In this review, we discuss the impact that these practical innovations have had on the study of rickettsiae. Prior to 1986, only eight rickettsioses were clinically recognized; however, in the last 10 years, an additional six have been discovered. We describe the different steps that resulted in the description of each new rickettsiosis and discuss the influence of factors as diverse as physicians' curiosity and the adoption of molecular biology-based identification in helping to recognize these new infections. We also assess the pathogenic potential of rickettsial strains that to date have been associated only with arthropods, and we discuss diseases of unknown etiology that may be rickettsioses. PMID- 9336674 TI - Management of asymptomatic term neonates whose mothers received intrapartum antibiotics--Part 1: Rationale for intrapartum antibiotic therapy. AB - The evaluation of the potentially septic newborn is often a source of frustration for practitioners. In the past, it has often been standard practice to evaluate and treat empirically all neonates whose mothers received antibiotics during labor, regardless of whether the infant had any signs or symptoms suggestive of infection. With the advent of recommendations for intrapartum antibiotic therapy to prevent early-onset neonatal group B streptococcal infections, this strategy is no longer practicable because too many infants would thus be evaluated and treated needlessly. This two-part review addresses the issues involved in managing asymptomatic newborns whose mothers received intrapartum antibiotics. This first part reviews the rationale behind strategies for preventing intrapartum transmission of bacterial infection. The administration of intravenous antibiotics to laboring mothers appears to reduce the incidence of group B streptococcal infections in neonates. Additionally, intrapartum antibiotic therapy for maternal chorioamnionitis may inhibit transmission of infection to the infant. Part 2--to be published separately--will address the evaluation and management of the newborn. PMID- 9336675 TI - The effect of prolonged bottle feeding on cow's milk intake and iron stores at 18 months of age. AB - Thirty-four toddlers were studied in a prospective, convenience sample comparison at their 18-month health supervision visit to examine the effect of prolonged (i.e., to 18 months of age) bottle feeding on both the daily volume of cow's milk intake and the toddler's iron stores (serum ferritin concentrations.) Seventeen toddlers had been weaned from the bottle by approximately 1 year of age, and 17 toddlers who remained on the bottle at 18 months of age were the compared group. The toddlers who remained on the bottle had significantly greater (P < 0.001) cow's milk intake (mean 26.3 oz vs 16.1 oz). The mean ferritin concentrations were lower in the persistent bottle group (17.3 micrograms/L vs 23.4 micrograms/L), but not significantly so. Questioning parents about their toddlers' continued bottle use at 18 months can provide a marker for potentially excessive cow's milk intake. PMID- 9336676 TI - Bioethical aspects of HIV infection in children. AB - The care of HIV-infected children is fraught with many bioethical conflicts and dilemmas that require careful attention if care is to be provided appropriately. Understanding of the interplay of such general principles as autonomy, nonmaleficence, confidentiality, and veracity helps to clarify the nature of specific conflicts. This article addresses both general principles and their specific applications to pediatric patients with HIV infection. It addresses these matters from the points of view both of patients and parents. It shows why conflict is practically inevitable, and it points the way toward prevention and resolution of conflict. Practical guidelines are provided in relation to the critical problem of disclosure of diagnosis to the patient. PMID- 9336678 TI - An analysis of morning report at a pediatric hospital. AB - The purpose of this study was to determine the types of cases residents select for morning report discussion and the educational value of postdischarge follow up of unknown cases. Between April and December of 1994, at Cardinal Glennon Children's Hospital in St. Louis, Missouri, random, resident, and group-selected patients listed at morning report were followed up throughout hospitalization. Patients were categorized based upon whether or not their morning report and discharge diagnoses were the same or different. Patients discharged without a diagnosis were followed up by chart review at 6 months to determine whether a diagnosis had been made. Data were analyzed by Chi-square analysis with Bonfferoni adjustment factor for multiple comparisons. Residents were more than two times more likely to select cases for discussion in which the diagnosis changed during hospitalization (P < 0.01). The 6-month follow-up yielded new diagnoses in only 21% of previously unknown cases. We concluded that residents do an exceptional job of selecting difficult diagnostic cases for discussion at morning report. Postdischarge follow up of unknown cases adds little new information for discussion at morning report. PMID- 9336677 TI - Pediatric morning report: an appraisal. AB - We examined and contrasted morning reports at two hospitals, university and community, that have a pediatric residency program. Patient diagnoses assigned at morning report were compared with final diagnoses to assess disease categories discussed and the value of including outpatient follow-up in this educational forum. Data were obtained during morning reports for 6 months by chief residents at university and private community hospitals. Pertinent history, physical examination, and laboratory and radiologic findings were recorded and were assigned a tentative morning report diagnosis based on morning report discussion. Cases were then reviewed at discharge and at 6 months to determine final diagnoses. At the university hospital, 58% of the cases were undiagnosed before presentation at morning report. Of those cases, 23% were assigned a diagnosis at morning report that differed from the final diagnosis. Similarly, at the private community hospital, 28% of cases were undiagnosed before presentation at morning report. Of those cases, 73% were assigned a diagnosis that differed from the final diagnosis. We conclude that the provision of follow-up at morning report is important for maximizing resident education. PMID- 9336671 TI - A primer on cytokines: sources, receptors, effects, and inducers. AB - Protection against pathogens is a prerequisite for survival of most organisms. To cope with this continuous challenge, complex defense mechanisms have evolved. The construction, adaptation, and maintenance of these mechanisms are under control of an extensive network of regulatory proteins called cytokines. A great number of cytokines have been described over the last 2 decades. This review consists of an overview of cytokines that are involved in immune responses and describes some historical and general aspects as well as prospective clinical applications. Major biological effects together with information on cytokine receptors, producers, inducers, and biochemical and molecular characteristics are listed in tables. In addition, some basic information is given on cytokine receptor signal transduction. Finally, the recent discoveries of cytokine receptors functioning as coreceptors in the pathogenesis of human immunodeficiency virus are summarized. PMID- 9336672 TI - Methicillin resistance in staphylococci: molecular and biochemical basis and clinical implications. AB - Methicillin resistance in staphylococci is determined by mec, composed of 50 kb or more of DNA found only in methicillin-resistant strains. mec contains mecA, the gene for penicillin-binding protein 2a (PBP 2a); mecI and mecR1, regulatory genes controlling mecA expression; and numerous other elements and resistance determinants. A distinctive feature of methicillin resistance is its heterogeneous expression. Borderline resistance, a low-level type of resistance to methicillin exhibited by strains lacking mecA, is associated with modifications in native PBPs, beta-lactamase hyperproduction, or possibly a methicillinase. The resistance phenotype is influenced by numerous factors, including mec and beta-lactamase (bla) regulatory elements, fem factors, and yet to be identified chromosomal loci. The heterogeneous nature of methicillin resistance confounds susceptibility testing. Methodologies based on the detection of mecA are the most accurate. Vancomycin is the drug of choice for treatment of infection caused by methicillin-resistant strains. PBP 2a confers cross resistance to most currently available beta-lactam antibiotics. Investigational agents that bind PBP 2a at low concentrations appear promising but have not been tested in humans. Alternatives to vancomycin are few due to the multiple drug resistances typical of methicillin-resistant staphylococci. PMID- 9336679 TI - The morning report. PMID- 9336670 TI - Helicobacter pylori. AB - Helicobacter pylori is a gram-negative bacterium which causes chronic gastritis and plays important roles in peptic ulcer disease, gastric carcinoma, and gastric lymphoma. H. pylori has been found in the stomachs of humans in all parts of the world. In developing countries, 70 to 90% of the population carries H. pylori. In developed countries, the prevalence of infection is lower. There appears to be no substantial reservoir of H. pylori aside from the human stomach. Transmission can occur by iatrogenic, fecal-oral, and oral-oral routes. H. pylori is able to colonize and persist in a unique biological niche within the gastric lumen. All fresh isolates of H. pylori express significant urease activity, which appears essential to the survival and pathogenesis of the bacterium. A variety of tests to diagnose H. pylori infection are now available. Histological examination of gastric tissue, culture, rapid urease testing, DNA probes, and PCR analysis, when used to test gastric tissue, all require endoscopy. In contrast, breath tests, serology, gastric juice PCR, and urinary excretion of [15N]ammonia are noninvasive tests that do not require endoscopy. In this review, we highlight advances in the detection of the presence of the organism and methods of differentiating among types of H. pylori, and we provide a background for appropriate chemotherapy of the infection. PMID- 9336680 TI - Infant botulism. PMID- 9336681 TI - Recognition of a paranasal sinus mucocele in a child with cystic fibrosis. PMID- 9336682 TI - Reversible pulmonary hypertension due to adenotonsillar hypertrophy in a patient for Fontan operation. PMID- 9336683 TI - Tuberous sclerosis with polycystic kidneys in an infant. PMID- 9336684 TI - Neonatal hemochromatosis. PMID- 9336685 TI - Biopsychosocial approaches to understanding human aggression: the first 30 years. AB - Theorists and researchers have long been aware of the potential utility of multidimensional explanations of human behavior, including human aggressive behavior. Biopsychosocial models are multidimensional explanations that attempt to provide a framework for understanding how biologic, contextual, and individual difference variables combine to influence human behavior. In this paper, the rationale for giving contemporary investigators a forum to discuss research findings on aggressive behavior from a biopsychosocial perspective is developed. The advantages and potential shortcomings of viewing aggressive behavior from a biopsychosocial perspective are briefly discussed. PMID- 9336686 TI - Biosocial bases of antisocial behavior: psychophysiological, neurological, and cognitive factors. AB - In this paper we review biosocial research and theory in the area of antisocial behavior. In particular, we focus on interactions between biological and social variables in predicting antisocial outcome. While many psychological researchers make statements concerning the potential importance of biosocial interactions, very few researchers actually test for such interactions in their data. The few studies that have reported biosocial interactions suggest that biological variables can protect against antisocial behavior in socially vulnerable individuals, and that social variables can protect against antisocial behavior in biologically vulnerable individuals. Further research is necessary to determine whether the effects of biosocial interactions on antisocial outcome are dependent upon particular biological or social factors. Preliminary evidence suggests that policy interventions that ameliorate the effects of perinatal risk factors could protect against antisocial and violent outcome. PMID- 9336687 TI - Human aggression in evolutionary psychological perspective. AB - This article proposes an evolutionary psychological account of human aggression. The psychological mechanisms underlying aggression are hypothesized to be context sensitive solutions to particular adaptive problems of social living. Seven adaptive problems are proposed for which aggression might have evolved as a solution--co-opting the resources of others, defending against attack, inflicting costs on same-sex rivals, negotiating status and power hierarchies, deterring rivals from future aggression, deterring mates from sexual infidelity, and reducing resources expended on genetically unrelated children. We outline several of the contexts in which humans confront these adaptive problems and the evolutionary logic of why men are cross-culturally more violently aggressive than women in particular contexts. The article concludes with a limited review of the empirical evidence surrounding each of the seven hypothesized functions of aggression and discusses the status and limitations of the current evolutionary psychological account. PMID- 9336688 TI - The relation between alcohol and aggression: an integrated biopsychosocial conceptualization. AB - The relation between acute alcohol consumption and aggressive behavior is a complex phenomenon that has been studied from a variety of different disciplines. This article reviews findings from both survey and experimental research. The influence of both situational and individual difference variables on the alcohol aggression relation is discussed and the strengths and weaknesses of various methodological approaches are highlighted. Current theoretical perspectives of the alcohol-aggression relation are reviewed. An integrated heuristic framework of the alcohol-aggression relation also is outlined. This conceptualization involves both distal and proximal risk factors for problems with alcohol and violence, which include biological, psychological, interpersonal, and contextual influences. Research and treatment implications of this framework are also discussed. It is recommended that researchers attempt to measure variables from a variety of domains in order to obtain a better understanding of this complex phenomenon. Furthermore, it is emphasized that there is a clear need to further implement and assess primary and secondary prevention efforts and to design integrated and flexible approaches for individuals with alcohol and violence problems. PMID- 9336689 TI - The serotonin hypothesis of aggression revisited. AB - Many contemporary theorists believe serotonin (5-HT) neurotransmitter functioning plays a role in the regulation of human aggressive behavior. We argue that the evidence supporting this 5-HT hypothesis of human aggression is less compelling than commonly assumed, due to (a) conflicting study results, and (b) significant methodological limitations of existing studies. Recent models that integrate the role of psychological and contextual variables in 5-HT--associated aggression are reviewed. The need to incorporate psychometrically sound measures of aggression in 5-HT studies, to use experimental and longitudinal designs, and to test hypotheses drawn from multifactorial models in future research is advocated. PMID- 9336690 TI - Role of hetero-enzyme complexes in acid-base balance. PMID- 9336691 TI - Soluble and membrane-bound forms of phosphate-activated glutaminase with different kinetic characteristics in pig and rat kidney mitochondria. PMID- 9336692 TI - The increase in glutamine synthesis from glucose caused by ammonium chloride in rabbit kidney tubules does not involve an increase in citrate synthesis. PMID- 9336693 TI - Stimulation of phosphoenolpyruvate carboxykinase gene expression in cultured LLC PK1-F+ cells. PMID- 9336694 TI - A novel Na(+)-dependent amino acid transporter with a high affinity to glutamine. PMID- 9336695 TI - Effect of K+ depletion on glutamate dehydrogenase. PMID- 9336696 TI - Characterization of the hepatic glutaminase promoter. PMID- 9336697 TI - Ensuring a minimum urine flow rate during water deprivation in chronic fasting. PMID- 9336698 TI - Role of adrenal steroids in stimulating ammonium excretion during acute metabolic acidosis. PMID- 9336699 TI - Mechanism(s) regulating matrix pH in rat kidney mitochondria. PMID- 9336700 TI - An overview of mitochondrial glutamine transport and phosphate-activated glutaminase in the kidney. PMID- 9336701 TI - Renal homocysteine metabolism. PMID- 9336702 TI - Metabolic requirement for inorganic phosphate by renal proximal tubules: influence upon L-glutamine metabolism. PMID- 9336703 TI - Hippurate as a source of renal and hepatic ammoniagenesis in different species. PMID- 9336704 TI - Regulation of hepatic protein degradation. PMID- 9336705 TI - No increase of ornithine concentration in the liver of ornithine transcarbamylase deficient spf-ash mice following intraperitoneal injection of ammonium chloride. PMID- 9336706 TI - Nitrogen metabolism in ammonium chloride-supplemented rats during protein depletion. PMID- 9336707 TI - How does the kidney handle plasma polyamines? PMID- 9336708 TI - Oxidation of glutamine in cancer cells and regeneration of adenylates via adenylosuccinate synthetase. PMID- 9336710 TI - Defective glycolysis and catabolism of protein and amino acids in skeletal muscle during metabolic acidosis. PMID- 9336709 TI - Leucine metabolism and protein dynamics in the human kidney. PMID- 9336711 TI - Incorporation of 15N-labelled precursors into urea isotopomers. PMID- 9336712 TI - RT-PCR localization of phosphoenolpyruvate carboxykinase in the proximal tubule of the rat during metabolic acidosis. PMID- 9336713 TI - On the relationships between the striatum and the pedunculopontine tegmental nucleus. AB - In this essay we consider the role of the pedunculopontine tegmental nucleus as a striatal output station. We review the relevant anatomical, electrophysiological, behavioral, and pathological studies and conclude that the pedunculopontine tegmental nucleus occupies an important position in striatal outflow, receiving motor output from the dorsal striatum and information from the ventral striatum relating to limbic processes of motivation and reinforcement. The hypothesis we present is that the pedunculopontine tegmental nucleus is at the very least an integral component of the limbic-motor interface, although in discussing this concept we also assess the likelihood that the limbic-motor interface is in fact a distributed system-that is, that limbic-motor interfacing is not all done by a single structure in the central nervous system but that different aspects of it are served by different systems. We present the hypothesis that the pedunculopontine tegmental nucleus is one critical site through which limbic information concerned with motivation, reinforcement, and the construction of novel associations can gain access to a stream of motor outflow coming from the caudate-putamen and directed toward pontomedullary systems without reference back to the cerebral cortex. This hypothesis is important because it highlights striatal outflow, which is not processed through the cortical re-entry systems, and also emphasizes the importance of pontine systems in cognitive processing. PMID- 9336714 TI - Taste and olfactory processing in the brain and its relation to the control of eating. AB - Taste processing in primates through the nucleus of the solitary tract and the primary taste cortex is shown to represent the identity and intensity of taste inputs. In contrast, in the secondary taste cortex in the orbitofrontal area, single neurons respond to the taste of a food only if hunger is present. The neurons here reflect the reward value of taste. They show sensory-specific satiety. In addition to neurons with best responses to sweet, salt, bitter, and sour, there are separate representations of the taste of protein ("umami") and of astringency (e.g., tannic acid). In the orbitofrontal cortex, olfactory inputs converge onto neurons with taste inputs, forming representations of flavor. Neurons in this region may also respond to the sight of food and to its texture. The olfactory representation for some neurons reflects the taste association of odors; and olfactory sensory-specific satiety is represented in this part of the brain. Rapid learning of visual-to-taste associations is also a feature of the neural processing that occurs in the orbitofrontal cortex. PMID- 9336715 TI - Psychobiology of HIV infection. AB - This review surveys evidence relevant to the proposition that psychobiologic factors may influence the progress of infection with human immunodeficiency virus Type 1 (HIV-1). Little research has directly examined the influence of psychobiologic factors on the pathogenetic mechanisms underlying HIV progression. However, basic research in neuroimmune interactions indicates that activation of the sympathetic nervous system and hypothalamic-pituitary-adrenal axis can influence several immunologic processes relevant to HIV pathogenesis and the body's ability to resist the progress of infection. A small number of observational natural history studies indicate that certain psychosocial characteristics may be associated with differential disease progression (e.g., subjective responses to highly threatening events, and inhibited psychosocial characteristics). We address some of the methodologic and conceptual issues critical to the interpretation of current results as evidence that psychobiologic processes influence HIV progression, and we conclude by highlighting promising areas for future inquiry. PMID- 9336716 TI - The primate substantia nigra and VTA: integrative circuitry and function. AB - A substantial amount of research has focused on the midbrain dopamine system and its role in mediating a wide range of behaviors. In diseases in which dopamine function is compromised, patients exhibit a constellation of symptoms suggesting that the dopamine system plays an important role in the integration of several functions. We have shown that there are subgroups of dopamine neurons that receive information from limbic and association areas and project widely throughout cortex and striatum, including motor areas. A dorsal tier of dopamine neurons receive input from the ventral (limbic) striatum and the amygdala and project widely throughout cortex. A more ventrally located group of dopamine cells receives input from both the limbic and association areas of striatum and project widely throughout the striatum including the sensorimotor regions. Through these projections the dopamine system can effect a wide range of behaviors. For the most part, structures of the basal ganglia are thought to be organized in parallel pathways. However, the behaviors affected by basal ganglia disorders can be in part explained by the integrative nature of the dopamine system and its links to motor, limbic, and association areas of the striatum and cortex. PMID- 9336717 TI - A Southwest Oncology Group perspective on the Revised European-American Lymphoma classification. AB - In recent years several new morphologic entities and a new classification system, Revised European-American Lymphoma Classification (REAL), have been proposed which affect the nomenclature and classification of lymphoid malignancies. This article reviews some of the features of the more common new entities, places these entities in immunologic context, explores the clinical utility of these entities, and provides a working clinical organization to the names. PMID- 9336718 TI - Pathologic prognostic factors in diffuse aggressive non-Hodgkin's lymphoma. AB - Recent advances in understanding the immunophenotypic, molecular genetic, and cytogenetic heterogeneity of diffuse large-cell lymphomas (DLCL) have provided new insights into the diversity of these disorders. Clinical prognostic factors are useful for predicting outcome in DLCL, but are surrogates of the underlying biology. The role of pathologic prognostic factors as the biologic correlates of clinical behavior in DLCL should allow the development of new prognostic models that incorporate both clinical and pathological data, and lead to improved outcome for those patients not cured by modern-day treatment regimens. PMID- 9336719 TI - Mistaken diagnoses of Hodgkin's disease. AB - This article reviews the frequency of and general types of diagnostic errors in Hodgkin's disease (HD) over the past several decades, discusses the most common diagnostic errors in the four histologic subtypes of HD today, and describes some of the clinical and pathologic features that may aid in avoiding a mistaken diagnosis of HD. PMID- 9336720 TI - Follicular lymphomas: do histologic subtypes predict outcome? AB - There has been a long history of debate as to whether histologic subtypes of follicular lymphoma are associated with unique outcomes. The controversy has been fueled by studies of small patient groups having heterogeneous prognostic factors followed for short intervals, and by a new proposal for the classification of lymphomas (REAL). The current report provides insight into the controversy by using a large group of patients of similar stage and treatment followed for up to 25 years. PMID- 9336722 TI - High-dose therapy and transplantation for low-grade lymphoma. AB - Because of the indolent natural history of low-grade lymphomas, long follow-up is needed to assess the overall success of high-dose therapy and autografting. Results to date suggest that patients with brief first or second remission with chemotherapy or those who attain only a partial remission with second-line or subsequent chemotherapy, but remain drug sensitive, may enjoy prolonged remissions relative to conventional treatment. The role of autografting in first remission is uncertain because the data from clinical trials are not mature. Furthermore, the risk of myelodysplasia after transplantation, the broad range of therapeutic alternatives available, and the option to give high-dose therapy upon relapse argue against autografting as primary treatment. Although purging the autograft of residual lymphoma as assessed by molecular methods has been associated with longer remission duration in one major center, the data from published series are remarkably similar, whether the graft was purged or not. Promising anecdotal data suggest that allogeneic transplantation for low-grade lymphoma deserves further study in prospective clinical trials. PMID- 9336721 TI - Overview of treatment of localized low-grade lymphomas. AB - The results of treatment of localized low-grade lymphoma have been reviewed with particular emphasis on the results of long-term follow-up of patients treated with radiation therapy at Stanford University. These data and results from numerous other centers suggest that 40% to 50% of patients with stage I and II follicular low-grade lymphomas can expect to achieve clinical cure of their disease with radiation therapy. Randomized trials published to date do not support the use of adjuvant chemotherapy. Although a policy of initial observation with deferral of treatment until the occurrence of disease progression is a well established approach to patients with advanced disease, no randomized studies exist that support this as a safe alternative to radiation therapy for early stage disease. New systemic therapies are required for the treatment of occult disease to prevent the occurrence of relapse outside of irradiated volumes. PMID- 9336723 TI - Long-term follow-up of platinum-based lymphoma salvage regimens. The M.D. Anderson Cancer Center experience. AB - Platinum-based regimens have been the most commonly used salvage therapy for non Hodgkin's lymphoma (NHL). In this update of two of these regimens with long-term follow-up data, we provide evidence that the etoposide-containing regimen (ESHAP) is superior to a cisplatin-cytosine arabinoside-based regimen (DHAP) in response rate, survival, and time to treatment failure. In spite of this superiority, the long-term outcome for most patients with relapsed or refractory NHL remains unfavorable. Newer salvage regimens such as MINE-ESHAP, or, when feasible, high dose chemotherapy with transplant, are probably better choices than either DHAP or ESHAP alone. PMID- 9336724 TI - Treatment considerations in the elderly patient with lymphoma. AB - One half of all non-Hodgkin's lymphomas occur in the elderly, a growing segment of the population in North America. Significantly, the incidence of lymphoma in general, and especially in the elderly, is rising rapidly. These trends will combine to double the number of cases of lymphoma in the elderly in the next 2 to 3 decades. Certain lymphomas can be treated as effectively in the elderly as in the young, and others are treatable but with only half the expectation of cure. Further improvements will be made as future clinical investigation focuses on the elderly with lymphoma as a special group. PMID- 9336725 TI - Treatment of organ transplant-related lymphoma. AB - First described in organ transplant recipients in 1968, post-transplant lymphoproliferative disorder (PTLD) remains an often devastating complication of immunosuppression. Similar, if not identical, Epstein-Barr virus (EBV)-associated B lymphoproliferations have since been described in congenital and other acquired immunodeficiency states. Although PTLD is often morphologically indistinguishable from aggressive non-Hodgkin's lymphoma, the pathogenesis, presentation, clinical course, and management options differ significantly from those of classic NHL. PMID- 9336726 TI - Alternative (non-P-glycoprotein) mechanisms of drug resistance in non-Hodgkin's lymphoma. AB - Drug resistance remains a significant obstacle to improving therapeutic outcome following treatment for malignant lymphoma. Eliminating P-glycoprotein as one mechanism of drug resistance may select for alternative, non-P-glycoprotein mechanisms of drug resistance. Understanding these alternative forms of drug resistance is imperative in order to improve therapy for NHL. PMID- 9336727 TI - The potential for immunoconjugates in lymphoma therapy. AB - Contemporary combination chemotherapy offers curative treatment for 30% to 50% of patients with advanced-stage, aggressive non-Hodgkin's lymphomas (NHL). However, conventional therapies cure few patients with indolent lymphomas or relapsed lymphomas of any histology. Myeloablative chemoradiotherapy with bone marrow or stem cell transplantation can provide pro-longed disease-free survival for a minority (20% to 50%) of patients with relapsed NHL, but new treatment approaches are clearly needed. In recent years, several groups of investigators have provided preliminary evidence suggesting that monoclonal antibodies (mAbs), in unmodified form or conjugated to toxins, drugs, or radioisotopes, may offer another effective therapeutic modality for patients with relapsed lymphomas. This article reviews current immunotherapy of NHL using antibody immunoconjugates. PMID- 9336728 TI - Targeted therapy for lymphoma with peptides. AB - Using the newly developed combinatorial peptide library methods, D-amino acid containing peptides that are specific against pan-B cell markers such as CD19, CD20, and CD22 can potentially be identified. These peptides can then be used as targeting agents for human lymphoma. PMID- 9336729 TI - Polychlorinated dibenzo-para-dioxins. PMID- 9336730 TI - Polychlorinated dibenzofurans. PMID- 9336731 TI - Decision support system to assist mechanical ventilation in the adult respiratory distress syndrome. AB - This paper presents a knowledge-based decision support system to assist mechanical ventilation in patients with the Adult Respiratory Distress Syndrome (DSSARDS). The knowledge base uses clinical algorithms developed from interviews and seminars with experts. The system contains 140 rules, applies backward chaining and was built on an IBM-PC compatible microcomputer. Clinical and physiological data and ventilator settings were used for suggestions of ventilatory support mode (VSMODE) and settings (MVSET) and for hemodynamic evaluation and therapy (HEMO). Success rates (s) and kappa coefficient (k) were used to measure agreement between DSSARDS and physicians at 4 decision steps related to: beginning of mechanical ventilation (FIRSTSET), VSMODE, MVSET and HEMO, DSSARDS prototype was evaluated in a development phase with 6 patients aged 48.6 +/- 15.9 years. Agreement results for 142 decision steps were: FIRSTSET k = 0.90, s = 0.93; VSMODE k = 0.76, s = 0.92; HEMO k = 0.58, s = 0.70, MVSET k = 0.86, s = 0.92 (p < 0.05 for all k). Improvements in the knowledge base were performed mainly in HEMO and VSMODE modules. The subsequent test phase studied 5 patients aged 54.8 +/- 11.0 years in a total of 900 decision steps. Results were: FIRSTSET k = 0.93, s = 0.95; VSMODE k = 0.93, s = 0.96; HEMO k = 0.97, s = 0.99, MVSET k = 0.96, s = 0.97 (p < 0.05 for all k). The results indicate significant agreement between DSSARDS and physicians for all decision steps. This suggests that DSSARDS may be used as a support for decision making and a training tool for mechanical ventilation in patients with the adult respiratory distress syndrome. PMID- 9336732 TI - The effect of surgical ICU triage patterns on differing severity adjusted outcomes in France and the United States. AB - INTRODUCTION: Surgical patients treated in French intensive care units (ICU's) appear to have higher mortality rates than patients in the United States. We hypothesized that this may be due to the French practice of not transferring dying patients from the ICU. We wished to determine if the different mortality rates could be explained by transfer practices for dying patients or other factors such as severity of illness. METHODS: Flowsheet data for 6,787 consecutive surgical ICU (SICU) patients from our institution over a 31 month period was entered into an ICU Clinical Information System which calculated the Day 1 Simplified Acute Physiology Score (SAPS) for each patient upon admission to the SICU. SICU and overall hospital mortality data were matched with severity data and the complete data set was analyzed against results for 2,604 surgical patients in French ICU's. Since terminally ill patients in France are not transferred to floor care, we also compared the French ICU mortality rate with both our SICU mortality rate and combined SICU and surgical floor mortality rates. RESULTS: Our overall SICU mortality was 1.7% and our combined SICU and hospital mortality was 4.2%, while the French ICU mortality was 14.1%. The French ICU's had more patients with higher severity of illness as measured by SAPS. When the effects of ICU transfer practices and severity of illness were considered, there were no mortality differences seen among patients admitted to the different units after elective surgery. Significant differences in mortality were seen when patients admitted emergently were studied. CONCLUSIONS: The differences in severity adjusted ICU mortality between French ICU's and our SICU are explained by different triage practices for terminally ill patients following elective ICU admission. These triage differences do not fully explain the mortality differences seen among patients emergently admitted to the ICU. Other factors such as the presence of trauma, ICU staffing practices, patient mix or other unidentified factors may be responsible for the severity adjusted differences in mortality among emergency surgical ICU patients. PMID- 9336733 TI - Spectral analysis of AC and DC components of the pulse photoplethysmograph at rest and during induction of anaesthesia. AB - We examined spectral components of beat to beat variability in AC and DC signals of the reflectance photoplethysmograph at finger and earlobe sites in 20 resting volunteers and 20 patients during propofol, alfentanil, isoflurane, nitrous oxide anaesthesia. We observed that at rest, the majority of spectral power at both sites and in both signals was in the low 'thermoregulatory' frequency band (0.01 0.08 Hz). These fluctuations were greater in the finger than in the earlobe and in the AC signal compared to the DC. With anaesthesia, low as well as mid (0.08 0.15 Hz) frequency variability decreased at both sites and in both signals whereas high frequency 'ventilatory' power (0.15-0.45 Hz) was maintained. During anaesthesia we found no significant differences between the spectral components of the AC or DC signals or between the finger and the earlobe sites. At all frequencies, the fluctuations in the AC and DC signals were out of phase with each other. PMID- 9336734 TI - Acoustic monitoring of the artificial airway--experimental results. AB - Non-invasive acoustic airway-monitoring was evaluated in an experimental study. Recording amplitude and travel time of acoustic pulse response, an acoustic pattern of airway's geometry was then calculated. Measurements on models and excised human cadaver lungs were performed to discover whether displacement or obstruction of the artificial airway could be detected by its acoustic equivalent. Regression analysis revealed a close correlation between displacement of tracheostomy tubes and the shifting of the acoustic area-distance function (corr. coeff.: 0.97-1) and an adequate correlation between acoustic and planimetrical determination of cross-sectional area within the tubes (corr. coeff.: 0.78). Dispersion analysis confirmed reasonable reliability of acoustic cross-sectional measurements (Coefficients of variation: 0.6-2.1%). The acoustic mapping thus provides an excellent approximation of the true displacement and/or obstruction of tracheostomy and endotracheal tubes. We conclude that acoustic monitoring may provide a helpful tool for achieving an early warning system of airway disturbancies in intubated and mechanically ventilated patients, as geometrical changes of airway configuration may be detected before they lead to relevant effects on respiratory metabolism. PMID- 9336735 TI - A preliminary laboratory investigation of air embolus detection and grading using an artificial neural network. AB - SUMMARY STATEMENT: Processed digitized Doppler signals abstracted from recordings during continuous air infusion in dogs were used to train a neural network to estimate air embolism infusion rates. BACKGROUND: Precordial Doppler is a sensitive technique for detecting venous air embolism during anesthesia, but it requires constant attentive listening. Since neural networks are particularly well suited to the task of pattern recognition, we sought to investigate this technology for detection and grading of air embolism. METHODS: Air was infused into peripheral veins of four anesthetized dogs at rates of 0.025, 0.05, 0.10, 0.25, 0.50 and 1.0 ml-1.kg-1.min-1 while digital recordings of the precordial Doppler ultrasound signal were collected. The frequency content of the recordings was determined by Fourier analysis. The output of the Fourier transform was the input to a neural network. The network was then trained to estimate the air infusion rate. RESULTS: The correlation coefficient between the size of the air embolism and the air infusion rate was greater than r2 = 0.93 for each of the four animals in the study when the network was trained using the data for all four dogs. When the data from a dog was withheld from the training set and used only for testing the correlation coefficients ranged from r2 = 0.75 to r2 = 0.27. For frequencies below 250 Hz, the acoustic energy tended to fall as the air infusion rate increased. The opposite occurred at frequencies above 325 Hz. CONCLUSIONS: Neural network processing of the precordial Doppler signal provides a quantitative estimate of the size of an air embolism. PMID- 9336736 TI - A simple method to calculate P50 from a single blood sample. AB - Hill's equation relating oxygen tension, saturation and P50 is used as the basis for a simple method to calculate P50 from a single blood sample. The effects of errors of measurement in oxygen tension and saturation are considered using the technique of sensitivity analysis. The method is illustrated using data published by Severinghaus. PMID- 9336737 TI - An interview with ICN's new President Kirsten Stallknecht. PMID- 9336738 TI - Mobilizing the public in support of quality nursing care. PMID- 9336739 TI - Specific activation of resting T cells against CA19-9+ tumor cells by an anti CD3/CA19-9 bispecific antibody in combination with a costimulatory anti-CD28 antibody. AB - Specific activation of resting lymphocytes for tumor targeting can be achieved by bispecific monoclonal antibodies (bi-mAb) with specificity for tumor antigens and T-cell-activating antigens, respectively, in combination with a costimulatory anti-CD28 antibody. We describe the generation and function of a bi-mAb with specificity for CD3 and for the tumor antigen CA19-9. The bi-mAb OKT3/NSI19-9 was generated by somatic fusion of two hybridoma lines secreting antibodies against CA19-9 and CD3, respectively. A hybrid/hybridoma was established, and its bi-mAb was characterized. In combination with a costimulatory anti-CD28 mAb resting peripheral lymphocytes could be activated specifically with T-cell proliferation and secretion of high amounts of interferon-gamma. On specific T-cell activation, bi-mAb OKT3/NSI19-9 could also redirect the cytotoxic effects of these T cells toward CA19-9+ tumor cells in vitro. Our results indicate that specific activation of resting T cells with bi-mAb OKT3/NSI19-9 in combination with an anti-CD28 mAb can activate resting T cells specifically and leads to antigen dependent bi-mAb-mediated cytotoxicity against CA19-9+ target cells. This approach may offer new perspectives for the specific immunotherapy of CA19-9+ tumors. PMID- 9336740 TI - Induction of antitumor immunity by intracerebrally implanted rat C6 glioma cells genetically engineered to secrete cytokines. AB - To test whether cytokine gene therapy can be applied to an immunologically privileged site, such as the brain, we investigated antitumor immunity in the brain induced by cytokine-secreting glioma cells. Three cytokine genes, interleukin-2 (IL-2), interleukin-4 (IL-4), and granulocyte-macrophage colony stimulating factor (GM-CSF) were transduced into a rat C6 glioma cell line via a retroviral vector, S2. Rats intracerebrally (IC) implanted with the C6 cells genetically engineered to secrete the cytokines, especially GM-CSF, manifested significantly higher survival rates than those with C6 cells or with C6 cells bearing the control vector (p < 0.002). In vivo, C6 tumors bearing the cytokine genes grew more slowly than wild-type tumors at any time point, and eventually diminished within 6 weeks after tumor cell implantation. Histopathological and immunohistochemical studies revealed that different cytokines induced diverse immune reactions. In the IL-2 group, CD4+ and CD8+ T cells dominated from day 3 to week 4, but disappeared at week 6. Some granulocytes were noted between weeks 2 and 4. In the IL-4 group, eosinophils were noted from day 3 to week 4, and CD4+ and CD8+ T cells, as well as macrophages at week 2. At week 6, only residual levels of macrophages and CD8+ T cells remained. In the GM-CSF group, granulocytes appeared as early as day 1 post-IC tumor implantation, and macrophages at day 2. CD4+ and CD8+ T cells were found from day 3 to week 4. At week 6, only residual CD4+ T cells and macrophages remained. Long-lasting antitumor immunity was confirmed in all groups by rechallenging surviving rats with wild-type C6 cells in the brain 100 days after implanting cytokine gene bearing C6 cells. In vivo depletion of GM-CSF by anti-GM-CSF antibody further confirmed that the immune reaction induced by GM-CSF-secreting tumor cells were mainly from the action of GM-CSF, rather than the immunogenicity of C6 cells. PMID- 9336741 TI - Transfection of IL-2 augments CTL response to human melanoma cells in vitro: immunological characterization of a melanoma vaccine. AB - We have transfected human melanoma cell line 518A2 with the cDNA encoding interleukin-2 (IL-2) or granulocyte-macrophage colony-stimulating factor (GM CSF), and compared cytokine-producing clones for their ability to induce melanoma specific cytotoxic T lymphocytes (CTL) from autologous peripheral blood mononuclear cells (PBMC) in vitro. The parental cell line expressed HLA-A1, HLA A2, ICAM-1, LFA-3, in addition to the common CTL antigens MAGE-1, MAGE-3, tyrosinase, gp100, and Melan-A/MART-1. Stimulation of autologous PBMC responders with the IL-2-transfected clone 518/IL2.14 specifically induced CTL lines reactive with all cell lines derived from the autologous patient. Strikingly, GM CSF-transfected 518A2 cells did not induce anti-tumor CTL reactivity. CTL induction against 518/IL2.14 was independent of HLA class II expression or CD4 help. The parental cell line 518A2 gained immunogenic properties when high concentrations of IL-2 were supplied exogenously, indicating that IL-2 produced and present at high levels locally by itself enhanced immunogenicity. From the autologous CTL line reactive with 518/IL2.14, clones were generated against an as yet unknown antigen, which was present in all autologous melanoma cell lines as well as in 7 of 15 HLA-A2+ melanoma cell lines tested, but not in melanocytes. These results will be discussed with respect to the possibility of using IL-2 transfected melanoma cells as a vaccine for treatment of patients with melanoma. PMID- 9336742 TI - Restoration of the immunocompetence by IL-2 activation and TCR-CD3 engagement of the in vivo anergized tumor-specific CTL from lung cancer patients. AB - The present study investigates the nature of the immunosuppressed state of the lymphocytes obtained from the malignant pleural effusion (effusion associated lymphocytes, EAL) of lung cancer patients. The immunocompetence of EAL from 13 patients was assessed by determining their T-helper cell phenotype, proliferative response to alpha CD3-activation, and their cytolytic activity against three tumor targets: the autologous tumor, Daudi, and K562. Flow cytometry analysis showed that the lymphocytes in EAL were predominantly T cells with < 1% natural killer cells. The T-helper cell phenotype was found to be predominantly of Th2 type, but could be readily converted to Th1 type by culturing the EAL in vitro, and this conversion was augmented by interleukin-2 (IL-2) or IL-2 plus alpha CD3. To test the cytolytic activity of EAL, it was found that after 6-day culturing, the EAL remained in an immunosuppressed state so that they failed to kill any of the three tumor targets. Stimulation with IL-2 partially restored the immunocompetence of EAL. Further engagement of TCR-CD3 by alpha CD3 fully restored the cytolytic activity of the EAL to kill the autologous tumor target but not Daudi or K562 tumor cells, and thus seemed to be tumor specific. The specificity was further confirmed by testing the activated EAL and normal donor peripheral blood lymphocytes against a variety of tumor targets and control targets. Furthermore, the killing by EAL against the autologous tumor target seemed to be major histocompatibility complex-restricted and was inhibited by anti-human leukocyte antigen class I antibody. The EAL from lung cancer patients also showed much reduced responsiveness to the alpha CD3 stimulation to induce proliferation, and addition of IL-2 restored the responsiveness. These results suggest that, through close contact with tumor cells, anergy of cytotoxic T lymphocytes (CTLs) was induced in vivo at a localized site. IL-2 stimulation and TCR-CD3 engagement could reverse the anergic state and restored the full competence of CTLs in EAL to mediate the specific anti-tumor killing against the autologous tumor. Proper manipulation of EAL may prove useful as a source of anti tumor effectors for cancer adoptive immunotherapy. PMID- 9336743 TI - Adoptive immunotherapy with tumor-cytotoxic macrophages derived from recombinant human granulocyte-macrophage colony-stimulating factor (rhuGM-CSF) mobilized peripheral blood monocytes. AB - We examined the mobilization of blood monocytes (MO) with recombinant human granulocyte-macrophage colony-stimulating factor (rhuGM-CSF) with regard to the in vitro generation of MO-derived tumor-cytotoxic macrophages (MAC) for use in adoptive immunotherapy of cancer patients. Eleven patients with progressing metastatic cancer received interferon (IFN)-gamma activated tumor-cytotoxic MAC generated in vitro from autologous MO with and without pretreatment with rhuGM CSF. RhuGM-CSF was administered subcutaneously at 10 micrograms/kg for 7 days. RhuGM-CSF treatment and adoptive cell transfer were well tolerated, and no toxicity greater than WHO II degrees occurred. Fever was the most common side effect and was seen in all patients during rhuGM-CSF treatment and during 9 of 22 MAC therapies. Bone pain was noted in 5 of 11 patients during rhuGM-CSF therapy. RhuGM-CSF treatment led to a continuous increase in the white blood cell counts and the number of MO within the leukapheresis products. The mean number of transfused MAC was 0.9 x 10(9) without rhuGM-CSF pretreatment and 1.9 x 10(9) after rhuGM-CSF administration. The maximum number of MAC that could be generated was 7.3 x 10(9), but after a dose escalation protocol only up to 2.7 x 10(9) MAC were transfused. Cytotoxicity against U937 cells increased during MO to MAC differentiation, but was decreased in both MO and MAC on treatment with rhuGM CSF. This study proves the feasibility of reinfusing MAC generated in vitro from rhuGM-CSF mobilized MO. PMID- 9336744 TI - Oncogene and cytokine expression of human colorectal tumors responding to immunotherapy. AB - Tumor samples from five patients with metastatic colorectal cancer who demonstrated tumor regressions in clinical trials of interleukin (IL)-1 beta, IL 2, and adoptive cellular therapy were analyzed for oncogene and cytokine mRNA expression. Tumors from eight nonresponding patients were also studied. Mutations of the ras protooncogene and overexpression of c-myc protooncogene were observed in both responding and nonresponding tumors. In contrast, none of the responding tumors expressed transforming growth factor (TGF)-beta 1 mRNA, whereas nonresponding tumors did. The expression of IL-1, IL-6, IL-8, IL-10, tumor necrosis factor-alpha, granulocyte macrophage-colony-stimulating factor, macrophage inflammatory protein (MIP)-1 alpha, MIP-1 beta, macrophage chemotactic protein, and RANTES was variable between responding and nonresponding patients. Although we cannot conclude that a pattern of oncogene and/or cytokine mRNA expression specifically characterizes sensitive colorectal cancers, these analyses-the assessment of TGF-beta 1 mRNA in particular-merit further evaluation as biomarkers prognostic of immunotherapy response. PMID- 9336745 TI - Characterization of tumor-infiltrating lymphocytes derived from human tumors for use as adoptive immunotherapy of cancer. AB - From 1991 to 1995, we initiated cultures of 94 fresh tumor samples of various histologies in an effort to grow tumor-infiltrating lymphocytes (TIL) using flasks and subsequent expansion in semipermeable bags. The five most prevalent tumor types from which TIL were successfully initiated were melanoma (25 successful initiates in 34 tumor samples, 74% success rate), colorectal cancer (12 of 18, 67%), renal cell carcinoma (9 of 12, 75%), breast (4 of 5, 80%), and sarcoma (5 of 7, 71%). The overall success rate for all tumors was 67 of 94 (71%). There were no instances of contamination from the time of culture initiation through harvesting of the final cell product for clinical use. The mean number of days to reach successful initiation (> 5 x 10(8) cells) was 35 +/- 24 days (mean +/- SD). TIL were then expanded from these successful initiates for either a repeated low-dose therapy (TIL reinfusion numbers of 5 x 10(8)-5 x 10(9) or for a repeated high-dose therapy (> 5 x 10(9)-5 x 10(10). The mean number of days to expand a TIL culture from the time of initiation to treatment for a first low-dose TIL was 59 days (range, 27-94 days) compared with 80 days (range, 33-209 days) for high-dose TIL. For patients who received a second or third high-dose TIL treatment, the average number of days needed to expand TIL was 39 days (n = 10) if there was no intervening cryopreservation of TIL, compared with 49 days (n = 10) if the culture had to be reestablished from cryopreserved TIL. For patients who received a second or third low-dose TIL, the mean number of days needed to expand TIL was 23 days (n = 3) if there was no intervening cryopreservation compared with 42 days (n = 17) if cultures had to be reestablished after cryopreservation of TIL. Low-dose TIL displayed predominantly CD4+ phenotype in 76% of 42 cultures, whereas high-dose TIL displayed predominantly CD8+ phenotype in 84% of 44 cultures. Cells bearing the natural killer (NK) phenotype (CD3-, CD56+) and the lymphokine activated killer (LAK) phenotype (CD3+, CD56+) were present in both low- and high-dose TIL cultures, but these phenotypes were never predominant. Cytotoxicity testing consistently demonstrated the persistence of NK and LAK activity in addition to the killing of allogeneic and autologous melanoma tumor targets. This work confirms that TIL cultures from most tumor types can be successfully established and expanded for therapeutic use, and repeated expansion from continuous TIL culture or cryopreserved TIL for repeated treatments is feasible. Such cultures are predominantly T lymphocytes that are phenotypically heterogeneous, and these phenotypes do not remain constant during prolonged time in culture. PMID- 9336746 TI - Biologic response modulation by tumor necrosis factor alpha (TNF alpha) in a phase Ib trial in cancer patients. AB - During a phase I study of recombinant human tumor necrosis factor (TNF) in cancer patients, serial immune studies were performed and analyzed for effects of TNF. The TNF (specific activity 9.6 x 10(6) U/mg protein, < 5.0 endotoxin units/mg protein) was given over 2 h intravenously on days 1, 8-12, 29-33, 50-54, and 71 75 at doses of 40, 80, 160, 200, and 240 micrograms/m2. Immunologic testing was performed before therapy three times and subsequently on days 2, 8, 10, 12, 29, 33, 50, 54, 71, 75, and off-study two times. Immune parameters evaluated included cytotoxicity [natural killer (NK), spontaneous lymphokine activated killer cells (LAK), LAK, and monocyte], cytokine production [spontaneous and stimulated interferon (IFN)-gamma and interleukin (IL)-2], superoxide production [resting and stimulated polymorphonuclear (PMN) and mononuclear cells (MNC)], and phenotype of peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD16, CD56, CD19). Data were analyzed for long-term effects, the effect after 1 day of treatment (day 1), and for weekly effect (change from day 1 to day 5 of a given treatment week). Significant decreases were seen in the spontaneous cytotoxicity of peripheral blood NK cells and IL-2-inducible LAK cells, whereas increases in spontaneous peripheral blood LAK activity were seen with TNF treatment. Consistent increases in superoxide production of resting PMN and MNC were demonstrated, with late increases in superoxide production by opsonized, zymosan treated PMN. No spontaneous IFN-gamma or IL-2 were noted in sera with treatment, but production of IL-2 by MNCs rose with TNF treatment. During 5 days of TNF treatment, the percentages of circulating CD8+ and CD56+ cells decreased, whereas that of CD4+ and CD19+ cells increased significantly and consistently, as determined by a multivariate analysis. Significant changes in several independently measured parameters were observed, including a dose-related diminished production of IFN-gamma by MNC stimulated by phytohemagglutinin and increased in vitro-generated LAK activity. Because there was no clinical response in this trial, no association of immunologic change with clinical response can be made. No biologically optimal dose of TNF was evident. The data suggest that TNF may act as a trigger cytokine, initiating a broad immune/inflammatory response. PMID- 9336747 TI - Inducing amenorrhea during bone marrow transplantation. A pilot study of leuprolide acetate. AB - OBJECTIVE: To evaluate, in a pilot study, the use and efficacy of a gonadotropin releasing hormone (GnRH)-agonist in inducing amenorrhea in women undergoing BMT. STUDY DESIGN: We evaluated the use of the GnRH agonist leuprolide acetate (LA) for the induction of amenorrhea in 10 postmenarcheal women prior to BMT. If there was a contraindication to the use of the intramuscular (i.m.) formulation of LA, the subcutaneous (s.c.) formulation was given as a daily intravenous (i.v.) bolus. Once the subject's platelet count was > 50,000/microL, the LA was discontinued. Menstrual bleeding, time from initiation of therapy to amenorrhea, and liver function test results were monitored. RESULTS: All subjects had induction of amenorrhea with the use of LA except for one subject with a large, myomatous uterus, who experienced light spotting. One subject who was thrombocytopenic at the prescribed time of the second dosage of i.m. LA received i.v. LA with documentation of continued pituitary/gonadal suppression. No adverse effects were determined to be directly related to either the i.m. or i.v. LA. CONCLUSION: LA is an option for the induction of amenorrhea in postmenarcheal women undergoing BMT. In thrombocytopenic subjects, administration of the s.c. formulation of LA by an i.v. route served as an alternative to i.m. injection and was documented to maintain gonadotropin suppression. PMID- 9336749 TI - Blood cultures in febrile patients after hysterectomy. Cost-effectiveness. AB - OBJECTIVE: To determine the impact and cost-effectiveness of blood cultures in patients with febrile morbidity following vaginal or abdominal hysterectomy. STUDY DESIGN: The charts of 192 abdominal and 150 vaginal hysterectomy cases performed at Walter Reed Army Medical Center for benign disease between June 1992 and June 1995 were retrospectively analyzed for postoperative febrile morbidity, fever evaluation, blood culture results, management and clinical course. RESULTS: Of the 342 hysterectomy cases reviewed, 46 (24%) abdominal and 16 (10.7%) vaginal hysterectomies had fever evaluations that included aerobic and anaerobic blood cultures. There were no blood cultures documenting bacteremia in the abdominal or vaginal hysterectomy patients evaluated for postoperative febrile morbidity. Two patients with positive blood cultures had preoperative infections and were eliminated from statistical analysis. Adherence to the definition of febrile morbidity eliminated another 19 (30.6%) of the 62 febrile patients phlebotomized for blood cultures. CONCLUSION: Blood cultures in the routine patient following vaginal or abdominal hysterectomy did not alter management and were not cost effective. Blood cultures may be appropriate for febrile patients with preoperative, intraoperative and postoperative risk factors for bacteremia. PMID- 9336748 TI - Laparoscopically assisted vaginal hysterectomy in a community hospital. AB - OBJECTIVE: To assess the impact and value of laparoscopically assisted vaginal hysterectomy (LAVH) in a community hospital. STUDY DESIGN: Recording and evaluating various clinical parameters related to LAVH. RESULTS: Our experience with 180 procedures confirmed that this procedure can be performed safely and effectively in a remote setting. The benefits are high patient acceptance, higher rate of vaginal as opposed to abdominal hysterectomies, less pain, shorter hospital stay and earlier return to daily activities. CONCLUSION: LAVH in a community hospital fills a need in gynecologic surgery. PMID- 9336750 TI - Treating adnexal masses. Operative laparoscopy vs. laparotomy. AB - OBJECTIVE: To compare operative laparoscopy vs. laparotomy for the treatment of adnexal masses. STUDY DESIGN: A retrospective review of all surgical cases who underwent operative laparoscopy or laparotomy for an adnexal mass during 1988 1995 at one multispecialty group practice. Preoperative screening for women over 45 included a CA-125 and ultrasound. If a malignant mass was encountered, it was immediately staged by laparotomy with the assistance of a surgical oncologist. During the study period 121 patients underwent ovarian cystectomy and 284 patients, oophorectomy. RESULTS: Laparoscopy was successfully completed in 118 of 127 (93%) oophorectomy and 71 of 72 (98%) of ovarian cystectomy patients. The incidence of malignant lesions at operative laparoscopy was 2%. The hospital stay for ovarian cystectomy was significantly shorter for laparoscopy (0.8 vs. 3.1 days). Hospital stay for oophorectomy was significantly shorter for laparoscopy (0.8 vs. 4.1 days). Ovarian cystectomy by laparotomy resulted in slightly more total complications than did laparoscopy (8% vs. 1%). Oophorectomy by laparotomy resulted in significantly more total complications than did oophorectomy by laparoscopy (29% vs. 3%). The mean total charge for laparoscopic oophorectomy was $5,873 versus $7,007 for laparotomy. The mean total charge for laparoscopic ovarian cystectomy was $4,507 vs. $5,541 for laparotomy. CONCLUSION: Treatment of adnexal masses by operative laparoscopy can be performed safely, with reduced morbidity and patient disability, and at a reduced cost. By having an oncologist backup in house, we have been able to convert most procedures to the laparoscopic approach. PMID- 9336752 TI - Neonatal cephalohematoma from vacuum extraction. AB - OBJECTIVE: To identify factors involved in the development of fetal cephalohematoma from vacuum extraction. STUDY DESIGN: Patients at > or = 34 weeks' gestation were randomly assigned to delivery by vacuum (n = 322) using the continuous (n = 164) or intermittent (n = 158) technique. Neonatal outcome with cephalohematoma was analyzed subsequently and related to prospectively recorded data. RESULTS: Approximately equal numbers of cephalohematoma were recorded in the two groups (continuous 20, intermittent 17; P = .686). Station at point of application (P = .008), increasing asynclitism (P < .001) and increasing application to delivery time (P = .002) correlated significantly with cephalohematoma. Only the last two factors achieved significance after stepwise multiple logistic regression analysis. Factors that did not achieve statistical significance were gestational age (P = .755), birth weight (P = .982), instrumental rotation (P = .896) and previous vaginal delivery (P = .051). CONCLUSION: In this prospective, randomized, controlled trial of vacuum-assisted delivery, the only predelivery factor found to predispose to neonatal cephalohematoma formation was increasing asynclitism. Although cephalohematoma formation was more likely to develop as the duration of vacuum application increased during delivery, only 28% of neonates exhibited this finding when the time from vacuum application to delivery exceeded five minutes. PMID- 9336751 TI - Adverse perinatal outcomes in young adolescents. AB - OBJECTIVE: To determine whether pregnant adolescents < or = 15 years of age developed more perinatal complications than older adolescents or adult women. STUDY DESIGN: We conducted a study of 147 nulliparous adolescents < or = 15 years of age who initiated prenatal care at the University of Texas Medical Branch at Galveston between January 1, 1992, and April 27, 1994. For purposes of analyses, these patients were then compared to two groups: (1) all nulliparas between the ages of 16 and 17 (n = 287) and (2) those 20-22 years old (n = 107) who delivered an infant of > or = 20 weeks' gestation and initiated care during the same interval at this same facility. Chi-square, Kruskall-Wallis or Student's t test initially was used to identify differences between groups in demographic characteristics and perinatal complications. Logistic regression analyses were then performed to determine whether observed differences in outcomes remained while controlling for potentially confounding variables. RESULTS: Adolescents < or = 15 years of age were more likely to develop anemia and less likely to deliver an infant who required admission to the intensive care unit. No differences were observed between groups in the prevalence of pregnancy-induced hypertension, preterm labor, preterm premature rupture of membranes, chorioamnionitis, meconium staining, endometritis, preterm delivery, low birth weight, low Apgar score or fetal demise. CONCLUSION: Adolescents < or = 15 years of age experience perinatal outcomes similar to those of older adolescents and young adult women. PMID- 9336753 TI - Cost of assisted reproductive technologies for a health maintenance organization. AB - OBJECTIVE: To calculate the cost of assisted reproductive technologies (ART) for a health maintenance organization (HMO), assess factors that contribute to the cost per delivery and to analyze how utilization rates can be controlled by the use of clinical criteria. STUDY DESIGN: Pregnancy outcome and a cost analysis of all ART cycles at an HMO in a state with mandated coverage for these procedures was performed. All patients (n = 148) undergoing ART cycles insured by the HMO performed at one in vitro fertilization (IVF) center during 1990-1995 were studied. RESULTS: ART cycle outcomes and a cost analysis, including global cycle and cancellation charges, medication costs, obstetric costs and neonatal care costs, were assessed. ART cycles (n = 375) included IVF (n = 278), gamete intrafallopian transfer (n = 46), cryopreserved embryo transfer (ET) (n = 42), zygote intrafallopian transfer/tubal embryo transfer (n = 7) and donor oocyte (n = 2). Pregnancy outcome with IVF was 18.3% deliveries per retrieval, for gamete intrafallopian transfer 27.8% deliveries per retrieval and for frozen ET 19% per procedure. Overall, 62/148 (41.9%) of the patients delivered. There were 35 singletons, 22 twin sets and 5 triplet sets. This resulted in an average cycle cost per delivery of $36,417. The mean obstetric and neonatal charges were $9,329 for a singleton delivery, $20,318 for twins and $153,335 for triplets. If these charges are expressed in terms of the number of infants born, a twin pregnancy would cost $10,159 per infant and a triplet pregnancy, $51,112. The ART cycle cost per HMO plan member was $2.49 per annum. Our IVF utilization was 295 cycles per million population. CONCLUSION: An HMO can control the cost of ART services by establishing preauthorization clinical criteria. Our utilization rates might be used as a benchmark for other insurers considering ART coverage. The cost of ART ($2.49 per annum) would be only a small fraction of the typical annual insurance premium. PMID- 9336754 TI - Venous flow through coiled and noncoiled umbilical cords. Effects of external compression, twisting and longitudinal stretching. AB - OBJECTIVE: To examine the association between umbilical cord coiling and perinatal morbidity. STUDY DESIGN: Ten umbilical cord segments, six coiled and four noncoiled, were categorized according to Strong's coiling index and were examined experimentally. The umbilical arteries and vein were perfused, pressurized, placed in a saline bath at 37 degrees C and subjected to compression, twisting and stretching while measuring venous flow. RESULTS: There was no statistically significant difference in umbilical venous flow between coiled and noncoiled cords when external compression, twisting and longitudinal stretching were applied to the cord segments. CONCLUSION: Differences in morbidity associated with umbilical cord coiling should not be attributed simply to mechanical factors, and other mechanisms should be sought. PMID- 9336755 TI - Second-trimester membrane rupture. Abortion induced with prostaglandin E2 after oxytocin failure. AB - OBJECTIVE: To test an effective method of terminating second-trimester pregnancy with ruptured membranes in women who fail to abort from an oxytocin infusion. STUDY DESIGN: Five patients with rupture of membranes during the second trimester of pregnancy and failed to abort using the traditional method of intravenous oxytocin infusion were treated with intrauterine instillation of prostaglandin E2 (PGE2) solution through a double-balloon device. RESULTS: All five patients aborted within 8.8 +/- 4.5 hours from the beginning of PGE2 instillation. No major complications occurred. The only side effect was short-duration pyrexia (less than 48 hours). CONCLUSION: Use of the double-balloon device and intrauterine instillation of PGE2 was effective for termination of pregnancy in patients with rupture of membranes who do not respond to oxytocin. PMID- 9336756 TI - Diagnosis and classification of diabetes mellitus: highlights from the American Diabetes Association. PMID- 9336757 TI - Nasopharyngeal teratoma. Report of a case with prenatal diagnosis and postnatal management. AB - BACKGROUND: Oral teratomas are rare entities. CASE: A patient was referred for elevations in the maternal serum alpha-fetoprotein. On ultrasound examination, a cyst was seen protruding from the fetal mouth. Postnatally, a teratoma was found to be filling the neonate's oral cavity. CONCLUSION: This case illustrates problems in determining the etiology of head and neck masses antenatally. It also shows that teratomas can appear to regress and yield positive amniotic fluid alpha-fetoprotein and acetyl-cholinesterase. The appropriate perinatal management of this potentially devastating disorder includes preparation for establishing an airway following delivery. PMID- 9336758 TI - Rare urogenital anomaly causing discharge and pain. A case report. AB - BACKGROUND: Patients with mullerian anomalies usually seek help because of poor reproductive performance. CASE: A 16-year-old, white woman presented with a persistent, brown vaginal discharge and right lower quadrant pain. Because of voluntary guarding, the uterus could not be palpated on bimanual examination. However, transvaginal sonography showed a right cystic mass and a slightly binodular contour of the uterus. Laparoscopic evaluation revealed the presence of a bicornuate uterus. A hysterosalpingogram showed a double uterine cavity and cervical canal below the right uterine cavity, leading to a cystic, paravaginal mass. Incision of the mass and drainage of its chocolate-brown fluid content revealed a small, blind vagina leading to a second cervix. Using a wire probe, an isthmic communication was demonstrated between the two uterine cavities, leading to the final diagnosis of bicornuate uterus, laterally communicating, with a double cervix and vagina, unilaterally blind. CONCLUSION: Awareness of the possibility of this clinically puzzling anomaly will avoid delayed or unnecessary surgical treatment. PMID- 9336759 TI - Abdominal wall endometriosis after amniocentesis. A case report. AB - BACKGROUND: Amniocentesis is a procedure performed commonly in the evaluation and early diagnosis of fetal chromosomal abnormalities. The procedure is safe, with a major complication rate of 0.5% and the most common minor complication, spotting. CASE: A 27-year-old woman underwent amniocenteses to document pulmonary maturity prior to cesarean section. She presented 18 months later with a 6-month history of left abdominal wall pain and a mass that enlarged 2 days prior to menses and shrank 1 week after. The mass was located at the prior amniocentesis site. Excisional biopsy revealed endometriosis on final pathology. CONCLUSION: To our knowledge, this is the first well-documented case of abdominal wall endometriosis following third-trimester amniocentesis. The procedure should be considered a possible complication of amniocentesis. PMID- 9336760 TI - Recurrent molar pregnancy after ovulation induction and repeat ovulation induction. A case report. AB - BACKGROUND: Although hydatidiform mole is not commonly encountered following ovulation induction, patients who have already had molar pregnancies are at increased risk of developing further molar diseases with worsening histologic characteristics. That fact underlies the ethical dilemma of repeat ovulation induction. CASE: A 38-year-old woman, gravida 3, para 0, had three consecutive episodes of hydatidiform subsequent to clomiphene citrate and gonadotropin ovulation induction. She seems to be the first in the literature to develop three consecutive molar pregnancies without a normal intrauterine pregnancy. CONCLUSION: Although ovulation induction commenced again in this patient after she gave informed consent, the risks underlying the ethical dilemma persist. PMID- 9336761 TI - Atypical Meigs' syndrome and bilateral ovarian stromal hyperplasia. A case report. AB - BACKGROUND: Atypical Meigs' syndrome has been observed in patients with dermoid tumors, struma ovarii, uterine leiomyomata and other benign pelvic tumors except for ovarian fibromas. Meigs' and atypical Meigs' syndrome present management decisions complicated by a high index of suspicion for malignancy. CASE: A 43 year-old woman, gravida 1, para 1, with ascites; a pleural effusion; radiologic evidence of enlarged, cystic adnexa; and a normal CA-125 level was found to have cortical stromal hyperplasia on bilateral ovarian pathologic evaluation. CONCLUSION: This is the first case of cortical stromal hyperplasia presenting with bilateral involvement of small ovaries, ascites and a pleural effusion. Meigs' syndrome and its variants develop with clinical pictures suggestive of malignancy. Thorough evaluation and individualized treatment are necessary. PMID- 9336762 TI - Conservative obstetric management of a gunshot wound to the second-trimester gravid uterus. A case report. AB - BACKGROUND: Gunshot wounds to the abdomen during pregnancy are becoming increasingly common. When the gravid uterus is injured but repairable and the fetus is either previable or dead, conservative management is an option, even if laparotomy is required for management of other injuries. CASE: A pregnant woman sustained a gunshot wound to the abdomen during the second trimester of pregnancy. The bullet injured the rectum and traversed the uterine cavity, with resultant rupture of membranes and direct, fatal injury to the fetus. Laparotomy was required for treatment of the maternal injuries; however, the pregnancy was managed conservatively. Spontaneous miscarriage occurred late on the second postoperative day. CONCLUSION: Conservative obstetric management is indicated in most such cases in which the fetus is previable or dead, although the clinician undertaking such management should be aware of the risks and possible complications. PMID- 9336764 TI - Lead-induced abnormalities in blood-brain barrier permeability in experimental chronic toxicity. AB - The aim of this paper was to determine whether prolonged drinking of lead acetate containing water by adult rats, which imitates environmental exposure to lead (Pb), affects some morphological and biochemical properties of rat brain microvessels. We noted a significant increase of lead level in capillaries and synaptosomes obtained from brains of rats under chronic toxicity conditions. Intravenously injected horseradish peroxidase (HRP) was used to evaluate the functional state of the blood-brain barrier (BBB). The results indicate that, systematically administered at low doses, lead induces BBB dysfunction. The changes, revealed in light microscopy and confirmed by electron microscopic studies, are typical for "leaky" microvessels, reported for variety of neuropathological conditions associated with BBB damage. Enhanced pinocytotic activity of the endothelial cells and the opening of interendothelial tight junctions, together with enormous phagocytizing action of the pericytes, are the most characteristic ultrastructural features noted. The presence of specific type of perivascular cells containing droplets of lipids in the cytoplasm, together with changes in phospholipid profile in brain capillaries, suggest that altered lipid composition of membranes may, at least in part, be responsible for changes in observed membrane permeability. PMID- 9336763 TI - Leiomyoma causing massive ascites, right pleural effusion and respiratory distress. A case report. AB - BACKGROUND: Pseudo-Meigs' syndrome, or atypical Meigs' syndrome, occurs when a pelvic mass other than an ovarian fibroma is present with hydrothorax and ascites. Leiomyomas rarely cause this condition. CASE: An otherwise healthy 31 year-old woman presented to the emergency department in acute respiratory distress with massive ascites, pleural effusion and a pedunculated leiomyoma. After receiving mechanical ventilation, she underwent myomectomy and recovered fully within four weeks. CONCLUSION: This unique presentation of pseudo-Meigs' syndrome should be included with malignancy in the differential diagnosis of a pelvic mass with ascites. PMID- 9336766 TI - Immunoblotting patterns of cytoskeletal dendritic protein expression in human neocortex. AB - Qualitative and quantitative evaluations of cytoskeletal proteins are critical for understanding physiological and pathological processes affecting the nervous system. Most of such studies on human samples have only used immunohistochemical techniques. We describe a complementary immunoblotting approach, for the assessment of neuronal cytoskeletal proteins, which employs fresh frozen postmortem tissues. We found that cytosolic fractions are suitable for qualitative and quantitative evaluations of four major dendritic cytoskeletal proteins: microtubule-associated protein (MAP)-2, MAP-5, and high- and medium molecular-weight nonphosphorylated neurofilaments. The enhanced chemiluminescence (ECL) technique revealed consistent and distinctive immunoblotting patterns for all four proteins in both monkey (no postmortem delay) and human (17-34 h postmortem interval) samples, some of which differed from those found in rodents. Quantitations of blots, by tissue protein-optical density curves that demonstrated linearity of the measurements in the 0- to 100-microgram range, support the feasibility of these immunoassays for the study of neurologic disorders. PMID- 9336765 TI - Downregulation of brain-derived neurotrophic factor mRNA in adult rat brain after acute administration of methylmercury. AB - Conventionally, assessment of the neurotoxicity of environmental pollutants relies on high-dosage treatment and nonspecific end points. In the present study, the early temporal and regional alterations in the mRNAs of neurotrophins were investigated following subtoxic doses of methylmercury (MeHg) in adult Sprague Dawley rats using in situ hybridization histochemistry and phosphoimaging evaluation. Decreases in brain-derived neurotrophic (BDNF) mRNA labeling intensities were seen in the dentate gyrus (DG; 44% of controls), and in the CA1 (72% of controls) and CA3c (70% of controls) cell layers of hippocampus after 8 mg MeHg/kg (ip) at 4 h, and at 1 h only in the DG. The decrease in BDNF mRNA expression in the DG was dose-dependent. At 3 d, regional levels had recovered. No significant changes could be detected in mRNA levels of the BDNF high-affinity receptor trkB or neurotrophin-3 mRNA at either 1 h, 4 h, or 3 d. Cresyl violet staining and GFAP immunohistochemistry did not reveal any major neuropathology in hippocampus at 2 wk. Thus, MeHg causes specific downregulation of BDNF mRNA, unlike many other perturbations of central nervous system homeostasis that have been shown to lead to upregulation of this mRNA. PMID- 9336767 TI - Lesioning of the inferior olive using a ventral surgical approach. Characterization of temporal and spatial astrocytic responses at the lesion site and in cerebellum. AB - Activated astrocytes, intrinsic components of both local and remote (axonal target regions) central nervous system injury responses, are now recognized as active metabolic and regulatory mediators in many neurological disorders. To further define these responses, we devised a new ventral surgical approach to unilaterally lesion the inferior olivary nuclear complex, which has a single predominant remote target, the cerebellum. Activated astrocyte number, volume, and density, as well as the total volume of brainstem involved in the astrocytic response, all peaked at postlesion day (pld) 4, returning toward, but not to, unoperated control values at pld 24 (p < 0.05). In contrast, the peak astrocyte response in the cerebellum was delayed, being greatest at pld 6 (p < 0.05 compared to control or pld 2). These responses were associated with increases in overexpression of S100 beta, an astrocyte-derived neurite growth factor, and with an increase in cerebellar steady-state levels of a neuronal injury response protein, the beta-amyloid precursor protein (beta-APP). This is similar to correlated increases in these two proteins that are found in epilepsy and Alzheimer disease. Our studies defining remote astrocytic and neuronal responses may be important for understanding glial-neuronal mechanisms underlying the spread of neuropathological changes in conditions such as Alzheimer disease. PMID- 9336768 TI - Suppression of drug-induced epileptiform discharges by cyclic AMP in rat hippocampus. AB - The effect of cyclic adenosine 3',5'-monophosphate (cAMP) on epileptiform activity in rat hippocampal slices was investigated. Bath-applied cAMP reversibly decreased the frequency of extracellularly recorded discharges in the CA3 subfield induced by bethanechol- or theophylline-containing solutions. Because cAMP was presumed to be relatively membrane impermeant, we developed and tested the hypothesis that this cAMP-mediated effect occurred extracellularly through the catabolic conversion of cAMP to 5'-AMP and, in turn, to adenosine, a known inhibitory neuromodulator. Three predictions derived from this catabolic hypothesis were tested. First, blockers of the enzymes involved were predicted to antagonize this effect of cAMP. In contrast, the coapplication of a cAMP phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX), or a 5' nucleotidase inhibitor, adenosine 5'-[alpha, beta-methylene] diphosphate (AMP CP), enhanced the cAMP-induced suppressive effect. Second, the nonhydrolyzable cAMP analogs, dibutyryl- and 8-bromo-cAMP, were predicted to be ineffective. Low concentrations (5-40 microM) of these two derivatives, however, also suppressed bethanechol-induced discharges, while, at a higher concentration (100 microM), both analogs increased discharge frequencies. Third, enzymatic catabolism of adenosine was predicted to antagonize cAMP's effect, but coapplying adenosine deaminase (10 U/mL) did not diminish this action. Because these data did not support the catabolic hypothesis, other, as yet undefined, mechanisms must be responsible for the discharge-suppressant effect of cAMP. PMID- 9336769 TI - Distinctive pattern of Bergmann glial pathology in human hepatic encephalopathy. AB - Alzheimer type II astrocytosis is the pathological hallmark of hepatic encephalopathy. These astrocytes undergo a characteristic morphological change and, in addition, lose immunoreactivity for glial fibrillary acidic protein (GFAP). However, a previous study in the portacaval shunted rat, a model of hepatic encephalopathy, revealed increased rather than decreased GFAP immunoreactivity in Bergmann glia, a specialized group of cerebellar astrocytes. In the present study, sections of cerebellar vermis from 15 cirrhotic patients with hepatic encephalopathy and varying degrees of Alzheimer type II astrocytosis were stained using antisera to GFAP. The Bergmann glial cells did not show altered GFAP immunoreactivity compared to controls. In addition, the degree of GFAP immunoreactivity was not correlated with the degree of Alzheimer type II change nor related to the aetiology of the liver disease. These results suggest a differential response of Bergmann glia in human hepatic encephalopathy. PMID- 9336770 TI - Complement and glutamate neurotoxicity. Genotypic influences of C5 in a mouse model of hippocampal neurodegeneration. AB - Using mice genetically deficient in the complement (C)-system component C5, this study explored a potential novel role of the C-system in Ca(2+)-mediated control of glutamate AMPA receptor functions. We found that Ca2+ preincubation of frozen brain tissue sections enhances AMPA binding capacity more dynamically in C5 deficient (C5-) than congenic C5 sufficient (C5+) mice. The Ca(2+)-mediated response was mostly localized to the CA3 and CA1 subdivisions of the pyramidal layers of the hippocampal formation. In C5- mice, kainic acid (KA) excitotoxicity that models hippocampal neurodegeneration abolished the Ca(2+)-mediated induction of hippocampal AMPA binding. The changes in AMPA binding preceded temporally and overlapped anatomically the appearance of apoptotic features in the same hippocampal neuron layers. C5- mice showed greater hippocampal neurodegeneration then C5+ mice. NMDA binding controlled for specificity of glutamate-mediated changes and found no C5 genotypic influences. The study gives further credence to the role of the C-system in modifying the intensity and outcome during response to conditions leading to hippocampal neurodegeneration. PMID- 9336771 TI - Short-term changes in NADPH-diaphorase reactivity in rat brain following perinatal asphyxia. Neuroprotective effects of cold treatment. AB - Perinatal asphyxia (PA) produces changes in nitric oxide synthase (NOS) activity in neuronal and endothelial cells of the striatum and neocortex. The changes were examined using a histochemical NADPH-diaphorase (NADPH-d) staining method. Newborn rats were exposed to severe PA at 37 degrees C and other groups were subjected to severe PA under hypothermic condition (15 degrees C) for 20 or 100 min, respectively. Quantitative image analysis was performed on the striatum and neocortex in order to count cell number of reactive neurons and to compare the pattern of staining between the different groups of animals. Severe asphyctic pups showed an important neuronal loss in striatum and neocortex that was reduced by hypothermia. NADPH-d(+) neurons with reactive processes were found in the lateral zone of the striatum and neocortex in asphyctic pups. Controls and hypothermic striatum showed rounded cells without reactive process, while no cells were stained in cortex. There was also an increase in NADPH-d activity in endothelial cells in severe asphyctic pups in striatum and neocortex vs control and hypothermically treated animals. Our data evidenced that an inappropriate activation of NOS in neuronal and endothelial cells induced by PA is related to neuronal injury. Hypothermia inhibits neuronal injury and may be a valuable neuroprotective agent. PMID- 9336772 TI - Encapsulated filtering blebs after trabeculectomy with mitomycin-C. AB - BACKGROUND AND OBJECTIVE: To determine the frequency of encapsulated blebs after guarded filtration procedures with mitomycin-C. PATIENTS AND METHODS: The authors reviewed the charts of all patients who had undergone a guarded filtration procedure with mitomycin-C. There were 235 patients (283 cases) who had more than 1 month of follow-up. RESULTS: An encapsulated bleb developed in 7 eyes (2.47%) of 6 patients. Identification of bleb encapsulation occurred at a mean follow-up time of 29.7 +/- 14.6 days after surgery. The mean intraocular pressure at that point was 24.2 +/- 13.5 mm Hg in the affected eyes. Three eyes were treated medically, and needling was performed in 4 eyes. CONCLUSION: There is a low frequency of encapsulated bleb formation after guarded filtration procedures with adjunctive mitomycin-C. PMID- 9336773 TI - Trabeculotomy combined with phacoemulsification and implantation of an intraocular lens for the treatment of primary open-angle glaucoma and coexisting cataract. AB - BACKGROUND AND OBJECTIVE: The authors previously reported the usefulness of trabeculotomy ab externo for the treatment of primary open-angle glaucoma in adult patients. In an attempt to elucidate the long-term risk-to-benefit ratio of this surgical modality in combination with cataract surgery, the authors conducted a retrospective study of the surgical effects and complications of a triple procedure: phacoemulsification, implantation (of an intraocular lens), and trabeculotomy (PIT). PATIENTS AND METHODS: The authors conducted a retrospective study of patients treated with PIT at multiple hospitals. Intraocular pressure (IOP) and visual function data were obtained from patients after PIT as an initial surgical treatment in cases where antiglaucoma medications failed to resolve uncontrolled IOP (higher than 21 mm Hg). Included in this study were 96 eyes of 64 patients with primary open-angle glaucoma and coexisting cataract. The mean follow-up period was 22.6 +/- 14.7 months (range 3-56 months). RESULTS: In 94 (98%) of the 96 eyes, the IOP was well controlled, having achieved a level of 21 mm Hg or lower at the final examinations. The mean preoperative IOP of the 33 eyes that underwent the triple procedure using a single flap method (PIT-I) was 24.3 +/- 3.9 mm Hg, with an average of 2.1 +/- 1.1 medications. At the final examinations, the mean IOP had dropped to 16.0 +/- 1.2 mm Hg, with an average of 1.2 +/- 1.2 medications. The mean preoperative IOP of the 63 eyes that underwent the triple procedure using a double flap method (PIT-II) was 26.2 +/- 6.2 mm Hg, with an average of 1.9 +/- 1.2 medications. At the final examination, the mean IOP for this group was 15.6 +/- 2.9 mm Hg, with an average of 1.0 +/- 0.9 medications. CONCLUSION: The long-term results from this multicenter study showed that the triple procedure, PIT, can be useful and effective as an initial surgical treatment for open-angle glaucoma in glaucoma patients with coexisting cataract. PMID- 9336774 TI - Fornix-based trabeculectomy with mitomycin-C. AB - BACKGROUND AND OBJECTIVE: Mitomycin-C (MMC) has been shown to improve the surgical success of trabeculectomy; however, the advantages of MMC have been evaluated almost entirely as an adjunct to limbal-based trabeculectomy. This study evaluated the efficacy and safety of fornix-based trabeculectomy with MMC for glaucomatous patients. PATIENTS AND METHODS: Between January 1993 and April 1995, 71 patients underwent fornix-based trabeculectomy with topical application of 0.4 mg/ml of MMC for 3 minutes. The conjunctiva-Tenon's capsule flap was spread over the limbus and sutured in order to create a visible crease with a water-tight closure. The mean follow-up time was 14.5 months. RESULTS: The mean intraocular pressure (IOP) before surgery was 32.4 +/- 9.7 mm Hg. The average postoperative IOP was 14.04 +/- 9.57 mm Hg. An IOP of 20 mm Hg or less was observed in 57 eyes (80%). Postoperatively, 37 eyes (52%) required no additional medical therapy. One month after surgery, only 2 patients had wound leakage with hypotony and choroidal detachment. Two eyes (3%) had suprachoroidal hemorrhage with loss of vision. A conjunctival "buttonhole" occurred in 2 eyes (3%), but only 1 persisted more than a month. CONCLUSIONS: Fornix-based trabeculectomy using intraoperative application of 0.4 mg/ml of MMC for 3 minutes was found to be as safe and effective as limbal-based trabeculectomy with MMC. PMID- 9336775 TI - The application of the macular photocoagulation study eligibility criteria for laser treatment in age-related macular degeneration. AB - BACKGROUND AND OBJECTIVE: To analyze the application of the Macular Photocoagulation Study eligibility criteria for laser photocoagulation of choroidal neovascularization, in view of the expansion of these criteria in recent years. PATIENTS AND METHODS: The authors prospectively analyzed 50 eyes of 47 consecutive patients with exudative age-related macular degeneration (AMD) to determine their suitability for treatment. RESULTS: Fifteen eyes (30%) were found to be suitable for laser photocoagulation. Patients eligible for treatment were more likely to have experienced visual symptoms for a months or less (P = .006), to have a visual acuity of 20/200 or better (P = .009), and to be younger in age (P = .02). Visual symptoms experienced for a month or less were more prevalent in extrafoveal exudative lesions compared with the subfoveal type (P = .01) CONCLUSIONS: Despite recent advances, laser photocoagulation still can be applied only to a minority of the patients with neovascular AMD. Prompt ocular examination following the onset of visual symptoms is essential. PMID- 9336776 TI - Management of silicone-induced cataract in AIDS patients treated for viral retinitis-associated retinal detachment. AB - BACKGROUND AND OBJECTIVE: To evaluate the outcome of cataract surgery for patients with acquired immunodeficiency syndrome (AIDS) who underwent vitrectomy and silicone tamponade for viral retinitis-associated retinal detachment. PATIENTS AND METHODS: The authors retrospectively reviewed the data of five AIDS patients (five eyes) who had cataract within a mean period of 4 months following vitrectomy and silicone oil tamponade for viral retinitis-associated retinal detachment. Phacoemulsification and implantation of a poly-methylmethacrylate posterior chamber intraocular lens were performed. The mean postoperative follow up was 3 months. RESULTS: Neither silicone oil loss nor retinal redetachment were reported postoperatively. Visual acuity improved in two eyes and remained unchanged in one eye. Total blindness occurred in two eyes. CONCLUSION: Although cataract surgery in these eyes is a relative easy procedure and does not interfere with the retinal status, visual outcome remains poor because of possible postoperative optic atrophy. PMID- 9336777 TI - Maximal, three-wall, orbital decompression through a coronal approach. AB - BACKGROUND AND OBJECTIVE: Only limited volume expansion is offered by traditional lateral orbital decompressions in which the anterior segment of the lateral wall is removed to allow lateral soft tissue prolapse. A great deal of additional soft tissue expansion can be obtained, not only laterally, but also posteriorly by removing the deep portion of the sphenoid wing. The authors report their experience in removing this bone through a coronal approach. PATIENTS AND METHODS: The authors performed maximal, three-wall, orbital decompressions through a coronal approach for 20 patients with thyroid-related orbitopathy. A disfiguring proptosis resulting from stable Graves' disease orbitopathy was the indication for surgery in all cases. Through a coronal approach, the lateral rim was left in place and thinned, augmented with specialized orbital rim onlay implants, or repositioned with osteosynthesis systems. The bone over the lacrimal fossa was sculpted to form a "keyhole" for the lacrimal gland, thereby providing additional orbital expansion. Once the medial canthal tendon and lacrimal sac had been elevated from their periosteal attachment, excellent exposure was obtained for medial and inferior orbital decompression. RESULTS: The authors report the results of 20 coronal orbital decompressions during a period of 44 months. Seven cases included lateral rim advancement. Up to 6 mm of retrodisplacement was achieved without rim augmentation, 9 mm with rim augmentation. DISCUSSION: The deep lateral orbital wall can provide significant room for volume expansion. The authors found that up to 6 mm of proptosis reduction can be obtained using the lateral wall alone. The coronal approach provides access to all four orbital walls for deep orbital decompression. The authors' philosophy of treatment in cases without compressive optic neuropathy is evolving toward the use of the lateral wall as the first approach with the incorporation of additional walls as needed. PMID- 9336778 TI - Indocyanine green videoangiography-guided laser photocoagulation of occult choroidal neovascularization. PMID- 9336779 TI - Ablation dynamics in laser sclerostomy Ab externo by means of pulsed lasers in the mid-infrared spectral range. AB - BACKGROUND AND OBJECTIVE: Sclerostomy ab externo with pulsed laser systems is currently in phase II clinical trials. The authors investigated the ablation dynamics of tissue treated with pulsed laser systems in the mid-infrared range to estimate the extent of thermo-mechanical damage to the sclera and the anterior chamber. MATERIALS AND METHODS: Freshly harvested porcine eyes were used. A bare 400-micron fiber in direct contact with tissue was used for fistulization. Polarization light microscopy, fast-flash photography, as well as optical and acoustic transients were performed for analysis. RESULTS: Substantial mechanical tissue deformation and dissections were found during pulsed laser ablation. The mechanical damage range within tissue far exceeds the pure thermal damage zone. Aspheric cavitation bubbles of up to 3 mm in length penetrate the anterior chamber after perforation. The cavitation demonstrates a significantly larger time constant in tissue than in water. CONCLUSIONS: Early fistula occlusions due to iris adherences may be attributed to iris trauma caused by cavitation. In response to the findings of this study, the authors propose an automatic feedback system to control the ablation process and minimize secondary ocular tissue effects. With respect to the overall damage zones, a new continuous-wave, mid infrared diode laser system seems to be superior to pulsed laser systems. PMID- 9336780 TI - Corneal blood staining following autologous blood injection for hypotony maculopathy. AB - Hypotony is a common complication following trabeculectomy in which antimetabolites are used. Autologous blood injection is an accepted form of treatment for hypotony that occurs secondary to overfiltration; however, injection into the filtering bleb has been associated with a rise in intraocular pressure for some patients with chronic postoperative hypotony. The authors describe a patient in whom corneal blood staining with raised intraocular pressure and loss of vision occurred as a result of autologous blood injection. PMID- 9336781 TI - Suture-supported posterior chamber intraocular lens implantation. AB - This article describes a new technique for secondary implantation of posterior chamber intraocular lenses in the absence of capsular support. In this technique, the intraocular lens is supported by a simple net, consisting of two mattress sutures that pass across the eye at the most anterior area of the pars plana ciliaris, 3 mm from the limbus. Both sutures have limbs 3 to 4 mm apart that intersect at 90 degrees angles in the pupillary area. The main advantage of this technique is that the sutures are placed prior to the opening of the globe, thereby reducing the time that the eye must be open for the insertion of the intraocular lens. Because the exposure time and the intraocular manipulations are decreased, the consequent vitreous disturbances and the risk of infection, cystoid macular edema, and retinal detachment are likewise reduced. Another advantage of this technique is the additional support for the intraocular lens, which reduces the incidence of decentration and tilt. Moreover, because the sutures are placed in the most anterior part of the pars plana ciliaris, which is avascular, the incidence of bleeding is decreased. PMID- 9336782 TI - Conjunctival autograft for pterygium surgery: how well does it prevent recurrence? PMID- 9336783 TI - ERCP-induced acute necrotizing pancreatitis: is it a more severe disease? AB - Acute necrotizing pancreatitis (ANP) is an uncommon but serious complication of endoscopic retrograde cholangiopancreatography (ERCP). This study compares the severity, clinical course, and long-term outcome of ERCP-induced ANP with ANP induced by other causes. A review of 72 consecutive patients with ANP treated surgically at the Mayo Clinic identified ERCP as the cause in 6 patients (8%). Compared to the remaining 66 patients, the post-ERCP group had higher APACHE II scores on admission (mean, 13 vs. 10) and more extensive pancreatic necrosis (mean, 55 vs. 47%). The post-ERCP group had a higher rate of infected necrosis (100 vs. 75%) and required earlier necrosectomy after the onset of pancreatitis (9 vs. 13 days). The rate of postoperative pancreatic and enteric fistulae was also higher (50 vs. 33%). Although the mortality rate in the post-ERCP group was lower (17 vs. 29%), they were significantly younger (50 vs. 62 years; p = 0.02) and all the survivors had residual long-term morbidity. ANP is more severe when ERCP-induced; infection introduced during the ERCP may, in part, account for this severity. PMID- 9336784 TI - Pleural effusion as a predictor of severity in acute pancreatitis. AB - Our objective was to determine whether pleural effusion is a predictor of severity in acute pancreatitis and, if so, whether it is an independent predictor. One hundred ninety-six consecutive cases of acute pancreatitis from October 1, 1994, to September 30, 1995, were reviewed. Medical records were analyzed for evidence of pleural effusion by chest radiograph and severe acute pancreatitis by identification of pancreatic necrosis or organ system dysfunction. Data were analyzed to determine if identification of pleural effusion provided an early sign of severity. Among 135 patients who underwent chest radiography, pleural effusion was seen in 16 of 19 (84.2%) with severe pancreatitis and 10 of 116 (8.6%) of patients with mild pancreatitis (p < 0.001). Pleural effusion was noted in severe pancreatitis prior to clinical or computed tomography evidence of severity in only 20% of cases. Pleural effusion is strongly associated with severity in acute pancreatitis but provides independent information on severity in only a minority of cases. PMID- 9336785 TI - Mismatch of duodenal deliveries of dietary fat and pancreatin from enterically coated microspheres. AB - Gastric emptying of dietary fat is affected by both chemical and physical factors; but when ingested as a free oil or an aqueous emulsion, fat may empty most rapidly immediately after the meal. In contrast, gastric transit of 1- to 3 mm spheres (like those of enterically coated pancreatins) is known to vary inversely with sphere diameter; and spheres leave the stomach initially slowly, if their diameter is > or = 1.6 mm. Our objective was to determine whether 2-mm microspheres of Pancrease would empty much more slowly than free or emulsified oil and whether 1.2-mm microspheres of Creon would empty as fast as free oil. We used a gamma camera to track the concurrent gastric emptying of 123I-labeled oil and 113mIn-labeled spheres of Pancrease or Creon in pancreatic-insufficient subjects with cystic fibrosis who ingested 20 g of free oil in spaghetti meals or 20 g of oil emulsified in a milk meal. We found that either type of oil emptied rapidly initially but slowed later, whereas either dosage form emptied slowly initially but rapidly later. Unexpectedly, the smaller spheres of Creon emptied about the same as Pancrease did after the spaghetti meal. For example, 50% of oil but < 25% of either dosage form had left the stomach by 90 min after the meals. Both dosage forms were lipophilic, forming aggregates in vitro. We concluded that the gastric emptying of either dosage form frequently lagged behind the emptying of oil from ordinary meals. We speculated that the similar transits of the 1.2-mm Creon and the 2-mm Pancrease resulted from aggregation of these microspheres in the presence of free oil. PMID- 9336786 TI - Effect of adenoviral early genes and the host immune system on in vivo pancreatic gene transfer in the mouse. AB - Gene transfer technology may provide a novel approach to treatment for pancreatic diseases. Recombinant adenovirus achieves efficient gene transfer in vivo. In this study, a murine model of adenoviral-mediated pancreatic gene transfer was developed, and the factors responsible for adenoviral elimination were investigated. Three days after direct pancreatic injection of a replication defective adenovirus containing the lacZ transgene, a high proportion (76.8 +/- 6.7%) of pancreatic cells expressed beta-galactosidase, the gene product. Gene expression was absent by 28 days posttransduction. In immunodeficient mice, beta galactosidase expression persisted with 20.0 +/- 6.0% of pancreatic cells staining positive 60 days after viral transduction. To test whether early viral proteins are the antigenic components responsible for the potent antiviral immune response, normal mice were injected with different adenoviral vectors containing early gene deletions. Vectors containing deletions in early region 2 or 4 expressed beta-galactosidase at 28 days. Presently available adenoviral vectors engineered to avoid this response offer minimal improvements in transgene duration. Further vector modifications or alternative strategies are needed to achieve stable pancreatic adenoviral transgene expression. PMID- 9336787 TI - Endocrine/exocrine intermediate cells in streptozotocin-treated Ins-IFN-gamma transgenic mice. AB - To trace the ontogeny of beta cell regrowth in adult transgenic mice that produce interferon-gamma in the islets (ins-IFN-gamma), their existing beta cells were depleted by treatment with high doses of streptozotocin (STZ). Initially, beta cell necrosis and degranulation were apparent in STZ-treated mice of both the BALB/c and the ins-IFN-gamma transgenic strains. The newly emerging transitional cells were then characterized by ultrastructural analysis. Interestingly, transitional cells harboring both exocrine and endocrine granules appeared frequently in ins-IFN-gamma transgenics after high-dose STZ treatment. New beta cells were produced primarily by the formation of new islets from the small pancreatic ducts. Beta cell regeneration in the ins-IFN-gamma transgenic mouse model is thus explained primarily by the budding of new islets from the ducts with acinar cells as possible precursors of islet cells. PMID- 9336788 TI - Inhibitory effects of the antiangiogenic agent TNP-470 on establishment and growth of hematogenous metastasis of human pancreatic carcinoma in SCID beige mice in vivo. AB - Effects on the liver of the antiangiogenesis agent O-(chloroacetyl-carbamoyl) fumagillol (TNP-470) on hematogenous metastasis of a human pancreatic carcinoma cell line were examined. One million PCI-43 cells, a human pancreas carcinoma cell line, were injected into the spleen of SCID beige mice, then TNP-470 at 30 mg/kg was administered subcutaneously every other day. The mice were killed 6 or 10 weeks thereafter and metastatic nodules in the liver were counted and measured microscopically. Metastases were inhibited and an approximately 10% loss of weight was evident in the TNP-470-administered mice. There was no suppression in maximal size of metastatic colonies in mice given the agent for 6 weeks, while inhibition was apparent in mice given the drug for 10 weeks. Suppression of proliferation and an increase in apoptosis were evident in metastatic nodules in the TNP-470-administered groups, following stainings for proliferative cell nuclear antigen and terminal deoxytransferase-mediated dUTP-biotin nick end labeling, respectively. TNP-470 inhibited the proliferation of human umbilical vein endothelial cells but not PCI-43 in vitro. TNP-470 did not suppress production of vascular endothelial growth factor by PCI-43 cells. Neovascularization in vivo induced by PCI-43 cells was suppressed in the TNP-470 administered mice, using a diffusion chamber placed in subcutaneous tissues of SCID beige mice. These observations suggest that inhibition of angiogenesis is effective in suppressing establishment and subsequent growth of hematogenous micrometastases of pancreatic adenocarcinoma to the liver. PMID- 9336789 TI - Specific regulation of pancreatic elastase I and II mRNA expression during postnatal development in the calf: reverse transcriptase-polymerase chain reaction analysis. AB - The specific regulation of pancreatic elastase I and II mRNA expression as well as of the protein, RNA, and DNA contents were determined during ontogeny in the calf. Specific activities and mRNA concentrations were quantified by spectrophotometry and reverse transcriptase-polymerase chain reaction, respectively. Calves were either milk-fed or weaned until slaughter at different ages. The biosynthesis of elastases I and II was modulated by postnatal development and weaning, leading to specific gene expression profiles. The levels of elastase I activity and of the corresponding mRNAs were found to evolve in a roughly similar way. On the contrary, elastase II activity level decreased sharply during postnatal development, while no changes were observed in the corresponding mRNA levels. After weaning, elastase I activity and mRNA levels, as well as elastase II mRNA levels, increased. However, the magnitudes of elastase I activity and mRNA inductions were different. Therefore, the expression of each gene in the calf pancreas is more or less independently regulated and the regulation is mainly pretranslational (elastase I) or translational (elastase II) during postnatal development and both pretranslational and translational at weaning. The translational efficiency of elastase I and II mRNAs might be influenced by the nature of dietary proteins. PMID- 9336790 TI - Duct epithelial cells cultured from human pancreas processed for transplantation retain differentiated ductal characteristics. AB - A procedure is described for the isolation and growth in vitro of epithelial cells from the duct network of human pancreas, referred to as DEC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (< 5 g) and, also, from the digest remaining after the isolation of islet cells from human pancreas, material that would normally be discarded. These were the only reliable sources for pancreas tissue available to us. This procedure shows that some of the techniques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the use of cholera toxin to prevent fibroblast growth and contamination obviates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclonal antibodies. The use of sieving to separate the digest immediately achieves a partial purification, which, coupled with that of allowing duct cysts to form, adds to the purity of the final preparation. The ductal system of the intact pancreas tissue and the DEC derived from it expressed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithelial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in otherwise waste pieces of human pancreas. It showed that these cells retained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo. PMID- 9336791 TI - Inhibitory effect of green tea extract on the process of pancreatic carcinogenesis induced by N-nitrosobis-(2-oxypropyl)amine (BOP) and on tumor promotion after transplantation of N-nitrosobis-(2-hydroxypropyl)amine (BHP) induced pancreatic cancer in Syrian hamsters. AB - Epidemiologic studies have shown a lower risk of gastrointestinal cancer in green tea drinkers. In the present study, the inhibitory effect of green tea extract (GTE) on the process of pancreatic carcinogenesis induced by N-nitrosobis-(2 oxypropyl)amine (BOP) and on tumor promotion after transplantation of N nitrosobis-(2-hydroxypropyl)amine (BHP)-induced pancreatic cancer were investigated in hamsters. In the first experiment, shortly after the initiation of pancreatic carcinogenesis by BOP, the animals in the GTE group were given GTE (0.5 mg/L) in their drinking water and the control group was given tap water. All animals were sacrificed 24 weeks later. There were no significant differences in body weight, water intake, or food consumption between the two groups during the experiments. GTE consumption was approximately 1.25 mg/day/100 g body weight during this experiment. Seven of the 13 hamsters (54%) in the control group were found to have pancreatic tumors, versus six of the 18 hamsters (33%) in the GTE group. The average number of tumors in the control group was 1.0/hamster, compared with 0.5/hamster in the GTE group. The overall incidence of macroscopic pancreatic tumors in the GTE group was about half that in the control group. The incidence of pancreatic cancer was 54% (12/13) in the control group and 44% (8/18) in the GTE group. The number of pancreatic cancers, including invasive carcinoma and carcinoma in situ, in the GTE group was 0.88/hamster, significantly lower than in the control group (1.68/hamster) (p < 0.05). The incidence of atypical ductal hyperplasia, which is thought to be an early pancreatic cancer, was also significantly lower in the GTE group than in the control group (1.50/hamster vs. 4.65/hamster) (p < 0.05). In the second experiment, 1-mm3 pieces of BHP-induced pancreatic cancer were transplanted into the back of hamsters. The control group (N = 16) was maintained on the basal diet and tap water throughout the experiment, and the GTE group (N = 16) was also maintained on the basal diet and tap water for the first 3 weeks after transplantation, when successful transplantation was confirmed and, thereafter, given tap water containing GTE (0.5 mg/L) for an additional 12 weeks. Tumor growth was similar in both groups until 11 weeks after transplantation, but inhibition of tumor growth became apparent after 11 weeks in the GTE group. At 13 weeks, the average tumor volume in the GTE group was 1.01 +/- 0.11 x 104 mm3, significantly smaller than that in the control group (1.98 +/- 0.37 x 104 mm3) (p < 0.05). The results demonstrated that GTE has an inhibitory effect on the process of pancreatic carcinogenesis and on tumor promotion of transplanted pancreatic cancer. These results suggest that GTE may come to serve as a chemopreventive and chemotherapeutic agent for pancreatic cancer. PMID- 9336792 TI - Cytotoxicity of peroxynitrite in rat pancreatic acinar AR4-2J cells. AB - Peroxynitrite (0.5-50 microM) induced dose-dependent cytotoxic effects in rat pancreatic acinar AR4-2J cells. Glutathione (2 mM) and ebselen (10 microM) partially reduced the cytotoxicity caused by 1-10 microM concentrations of peroxynitrite. Higher concentrations (10-50 microM) of peroxynitrite induced DNA smear suggestive of necrosis, while lower concentrations (2-5 microM) induced DNA fragmentations suggestive of apoptosis. The effects of peroxynitrite on [Ca2+]i showed a similar dose dependency. Peroxynitrite concentrations > 10 microM rapidly increased [Ca2+]i in a dose-dependent manner, while concentrations < 5 microM did not affect [Ca2+]i. In contrast, the presentation of wild-type P53 was accelerated at lower concentrations of peroxynitrite (< or = 10 microM) but not at higher concentrations (50 microM). The present study suggests that peroxynitrite at lower concentrations (2-5 microM) induces wildtype P53 and apoptosis, which is potentially a protective response toward the DNA damage caused by peroxynitrite. On the other hand, higher concentrations of peroxynitrite (10-50 microM) rapidly increase [Ca2+]i and eventually induce necrosis. PMID- 9336793 TI - Stimulation of exocrine pancreatic secretion by soybean trypsin inhibitor does not depend on the masking of luminal trypsin activity in rats that have bile pancreatic juice diverted into the ileum. AB - We reported previously that dietary proteins stimulate pancreatic secretion by a mechanism not involved in masking trypsin activity in rats that have bile pancreatic juice (BPJ) diverted from the proximal small intestine for 7 days. However, BPJ in the distal small intestine is possibly responsible for the stimulation of pancreatic secretion in chronic BPJ-diverted rats. To examine whether the BPJ-dependent mechanism operates in the distal small intestine of chronic BPJ-diverted rats, we investigated pancreatic responses after inhibition or removal of pancreatic trypsin activity in the distal small intestine of conscious rats. Duodenal instillation of soybean trypsin inhibitor (SBTI), which is a proteinaceous trypsin inhibitor, stimulated pancreatic secretion in the chronic BPJ-diverted rats, whereas a nonpeptidic trypsin inhibitor, FOY 305, did not. Ileal administration of both trypsin inhibitors did not enhance pancreatic secretion in the diverted rats. Exclusion of luminal BPJ from the distal small intestine was also ineffective in causing pancreatic exocrine secretion in the chronic BPJ-diverted rats. These observations reveal that ileal BPJ does not contribute to the stimulation of pancreatic secretion in rats that have BPJ chronically diverted into the ileum, as in intact rats, and that a duodenal instillation of SBTI stimulates pancreatic secretion as a protein, and not as a trypsin inhibitor. PMID- 9336794 TI - Cerulein-induced acute pancreatitis in rats--does bacterial translocation occur via a transperitoneal pathway? AB - Bacterial infectious complications are the most common cause of morbidity and mortality associated with acute pancreatitis. Most pathogens are common gastrointestinal flora, indicating that the gut is the source of pancreatitis related infections. However, the route whereby the microorganisms reach distant organs remains speculative. We tested the hypothesis that spread of bacteria occurs via a transperitoneal pathway. Acute interstitial pancreatitis (AIP) was induced in antibiotic (gentamicin, bacithracin, neomycin)-decontaminated rats by intravenous infusion of cerulein. Effects of pancreatic necrosis (PN) were studied in rats that received additional injections into the peritoneal cavity of pancreatic tissue obtained from donor rats. The rats were inoculated with Escherichia coli (O2:KN:H18) resistant to the antibiotics used for decontamination either orally (10(12) microorganisms; experiment I) or intraperitoneally (10(8) microorganisms; experiment II). Moreover, the rat peritoneal cavity wash was inoculated with 10(8) E. coli in vitro (experiment III). In rats with AIP and PN, recovery of the bacteria from liver, spleen, pancreas, lung, and blood following oral inoculation demonstrated that acute pancreatitis promotes bacterial translocation from the gut. The absence of E. coli in these organs following intraperitoneal inoculation showed that the bacteria do not spread from the peritoneal cavity. Rats with PN cleared E. coli from the peritoneal cavity in a shorter period than rats with AIP and controls (5 vs. 7 and 8 days; p < 0.05). The multiplication rate of E. coli in peritoneal cavity wash was lower in rats with PN than in rats with AIP and controls (p < 0.01). We conclude that (1) translocation of E. coli from the gut during cerulein induced acute pancreatitis occurs via nonperitoneal pathways, (2) the peritoneal cavity acts as a trap for the bacteria rather than a source of bacterial seeding, and (3) PN impairs survival of E. coli in the peritoneal cavity via inhibition of the bacterial multiplication in this model. PMID- 9336795 TI - Effect of Irsogladine on gap junctions in cerulein-induced acute pancreatitis in rats. AB - The capacity for intercellular communication (IC) via gap junctions is found in normal pancreatic acinar cells. The major role of IC is considered to be the maintenance of tissue homeostasis and the regulation of signal transmissions. Up to now, the participation of IC via gap junctions in acute pancreatitis has not been reported. We investigated the role of IC in cerulein (Cn)-induced acute pancreatitis in rats using irsogladine, an enhancer of IC via gap junction. Acute edematous pancreatitis was induced in rats by two intraperitoneal injections of 40 micrograms/kg Cn. Rats received various doses (25, 50, or 100 mg/kg body weight) of irsogladine orally, 15 and 2 h before the first Cn injection. The normal control group received only vehicle. The severity of pancreatitis was evaluated enzymatically and histologically 5 h after the first Cn injection. In Cn-induced acute pancreatitis, irsogladine significantly lowered the serum amylase level, the pancreatic wet weight, and the pancreatic amylase and DNA contents, in a dose-dependent manner. Particularly, the amylase content improved to the level of the normal controls. Histologically, the severity of pancreatitis was reduced significantly by treatment with irsogladine and no discernible vacuolization was seen in the group with 100 mg/kg irsogladine treatment. By immunofluorostaining pancreata with anti-connexin 32 (Cx32; a gap junction protein) antibody, we found that pancreatic acini were diffusely positive for Cx32 in the control group, but the number of Cx32-positive grains decreased markedly, to 19%, in the pancreatitis group. With 100 mg/kg irsogladine treatment, the number of Cx32 grains recovered to 70% of the normal control value. These findings indicate that IC via gap junction is disturbed in Cn induced pancreatitis, which may result in the breakdown of tissue homeostasis and the progression of acute pancreatitis. PMID- 9336797 TI - Stimulatory effect of synthetic luminal cholecystokinin releasing factor (LCRF) fragment (1-35) on pancreatic exocrine secretion in conscious rats. AB - The effects of a synthetic putative luminal cholecystokinin (CCK) releasing factor (LCRF) fragment (1-35) on pancreatic exocrine secretion were examined in conscious Wistar rats and a mutant strain of rats lacking the CCK-A receptor. Intraduodenal injection of graded doses of the LCRF fragment induced biphasic responses in Wistar rats. The injection of 0.1 microgram of the LCRF fragment produced a maximal response, while 1 microgram produced a lower response. No significant effect was observed in rats lacking the CCK-A receptor. The synthetic LCRF fragment stimulated pancreatic secretion via CCK-A receptors in conscious rats. PMID- 9336796 TI - Neutrophil behavior in pancreas and liver and the role of nitric oxide in rat acute pancreatitis. AB - The behavior of neutrophils in a rat acute pancreatitis model was observed in the pancreas and liver using fluorescence microscopy with an image analyzing system after labeling with a specific fluorescent reagent. Nonviable cells of both organs were also labeled and quantified. The role of nitric oxide in neutrophil accumulation and organ damage was estimated by administering a relatively selective inhibitor of constitutive nitric oxide synthase, N-nitro-L-arginine (L NNA). The animal model of acute pancreatitis was induced by cerulein injection (80 mg/kg). Two groups were created, one given and the other not given L-NNA (2.5 mg/kg) prior to the induction of pancreatitis. The number of accumulated neutrophils in the pancreas and liver increased in a time-dependent manner. There was a close relation between the distribution of the neutrophils and inviable acinar cells or hepatocytes. When pretreated with L-NNA, the numbers of accumulated neutrophils and nonviable cells increased significantly in the pancreas. In the liver, a more pronounced accumulation of neutrophils was observed after treatment with L-NNA. Although hepatocyte injury was mild despite the neutrophil accumulation in the control, such injury was marked in the group treated with L-NNA. This suggests that neutrophils serve an important role in exacerbating acute pancreatitis and that nitric oxide provides a defense mechanism against neutrophil accumulation in pancreas and liver. PMID- 9336798 TI - Long-term blockade of cholecystokinin (CCK): effects of L 364,718 (a CCK receptor antagonist) on pancreatic enzyme storage and secretion. AB - To determine the effect of long-term blockade of cholecystokinin (CCK) on both pancreatic storage and secretion processes, L 364,718 (a CCK receptor antagonist) was administered to rats at 0.1 mg/kg/day for 3, 7, and 15 days. Zymogen granules were analyzed by flow cytometry to determine their light scattering properties forward scatter and side scatter-as well as their amylase content measured by a specific antiserum. The mean number of zymogen granules per cell was counted on pancreatic sections using electron microscopy. DNA content, pancreatic weight, and enzyme secretion were also studied under both basal conditions and CCK infusion at a dose of 1.25 micrograms/kg/h, which is able to displace the CCK receptor antagonist. Two subpopulations of zymogen granules (Z1 and Z2) were identified on the basis of their light scattering parameters, in both control and L 364,718-treated rats. L 364,718 administered for 3, 7, and 15 days induced a significant reduction in the amylase content of individual zymogen granules, for both Z1 and Z2 zymogen granule subsets. In contrast, the number of zymogen granules per cell increased from day +3 of treatment onward, the highest values being detected at day +7. Hyperplasia was observed only at day +15. Basal enzyme secretion decreased significantly in rats treated with L 364,718 for 3 and 7 days but recovered to control values after 15 days of treatment. No significant differences in CCK-stimulated amylase secretion were observed between control and L 364,718-treated rats. At day +15 of L 364,718 treatment a significant increase in enzyme secretion was observed with respect to shorter treatment periods; this was associated with a significant increase in both the number of cells and the number of zymogen granules per cell. Our results indicate that chronic administration of L 364,718 induces a biphasic effect on pancreatic function. Interestingly, although enzyme secretion reached recovery after long-term treatment (15 days), the storage process is altered since the mean enzyme content in each individual zymogen granule remains significantly reduced. PMID- 9336799 TI - A new experimental model for primary chronic pancreatitis. PMID- 9336800 TI - Common association of HPV 2 with anogenital warts in prepubertal children. AB - Anogenital (AG) warts in 31 prepubertal children were HPV typed by nonisotopic in situ hybridization (NISH) using digoxigenin-labeled probes for human papilloma virus (HPV) types 1-5, 6, 11, 16, 18, 31, and 33. Mode of transmission was determined from historical, clinical, and laboratory data independent of HPV typing. HPV 2 was detected most commonly (13/31 warts) followed by HPV 6 (7/31), HPV 11 (5/31), and HPV 16 (1/31). Although not reaching statistical significance, our results suggested that a mucosal HPV type (6, 11, 16) in a child's AG warts implied transmission from mucosal warts and conversely cutaneous HPV 2 transmission from warts at a cutaneous site. HPV typing provided no helpful information regarding actual mode of transmission of AG warts in these children. The high prevalence of HPV 2 in children's AG warts and the low prevalence of sexual abuse (2 of 31 children) found in this study suggest innocent auto- or heteroinoculation from cutaneous warts may be a common means by which children acquire AG warts. PMID- 9336801 TI - Infraorbital crease and atopic dermatitis. AB - The usefulness of a prominent infraorbital skin crease as a marker of atopic dermatitis (AD) was examined in 500 consecutive school children 3 to 11 years of age. Infraorbital crease was recorded by two trained observers according to a strict protocol, and AD was determined by an independent dermatologist who was blinded to the study design. A prominent infraorbital crease was present in only 4 of 20 (20%) children with AD, compared with 171 of 480 (35.6%) children who did not have AD (p > .05). While infraorbital crease may be of some use in diagnosing individual cases of AD in a hospital setting, it appears to be less useful in population-based studies because of its poor validity and repeatability. PMID- 9336803 TI - An epidemiologic study of perianal dermatitis among children in Egypt. AB - Perianal dermatitis is a common problem occurring among infants and children. Streptococci, particularly beta-hemolytic group A organisms, play a major role in its causation. An epidemiologic association between perianal dermatitis caused by group A beta-hemolytic streptococci in some patients and pharyngeal colonization with the same organisms seems to exist. A similar relation is also true for other organisms, including non-group A beta-hemolytic streptococci and Staphylococcus aureus. This was the main conclusion of a hospital-based study performed on 150 children with perianal dermatitis. All patients were subjected to a questionnaire, clinical examination, two perianal swabs, and two throat swabs. The bacteriologic examination of the perianal swabs revealed the presence of beta hemolytic streptococci in 35.3% of the cases, half of which were of the group A beta-hemolytic strain (17.3%) and half of which were non-group A (18%). Throat swabs revealed the presence of beta-hemolytic streptococci in 44% of cases, half of which were found to belong to group A (21.3%) and half to non-group A (22.7%). Among patients with perianal dermatitis caused by group A beta-hemolytic streptococci, 53.8% had associated pharyngeal colonization by the same organism. S. aureus was isolated from the perianal skin in five patients (3.4%); in four of whom the same organism also grew in cultures from throat swabs. A relatively good association between pharyngeal colonization by beta-hemolytic streptococci and Staphylococci and the presence of perianal dermatitis caused by the same organisms was demonstrated using the Kappa test of agreement (K = 0.4). PMID- 9336802 TI - Menkes' kinky hair syndrome: ultrastructural cutaneous alterations of the elastic fibers. AB - Menkes' kinky hair syndrome is associated with the defective functioning of several copper-dependent enzymes due to impaired copper absorption, transport, or metabolism. Lysyl oxidase is a copper-requiring enzyme that catalyzes the oxidative deamination of lysyl residues linking two adjacent chains of tropoelastin polypeptides into an insoluble network. Elastin of the connective tissue is the responsible protein for the elastic properties of the skin. We report transmission electron microscopy findings concerning elastic fiber alterations of the skin in three patients with Menkes' syndrome. The reticular dermis showed marked changes in the elastic fibers with a paucity of the central amorphous component while retaining normal microfibrillary material. These ultrastructural observations, to the best of our knowledge, are reported for the first time in skin from these patients and may be readily interpreted in terms of a specific biochemical defect in elastogenesis. PMID- 9336804 TI - Autosomal recessive inheritance of erythrokeratoderma variabilis. AB - Erythrokeratoderma variabilis is a rare genodermatosis conventionally regarded as autosomal dominant in inheritance. We describe the clinical features and light and electron microscopic findings in two affected siblings born to unaffected parents and suggest an autosomal recessive mode of inheritance in this family. We also briefly review the literature on this disorder. PMID- 9336805 TI - Epidermolysis bullosa associated with congenital localized absence of skin, fetal abdominal mass, and pyloric atresia. AB - A 2320-g male infant was delivered at 35 weeks gestation to a mother who had polyhydramnios. He had a combination of congenital localized absence of skin, unilateral hydronephrosis, and hydroureter due to ureterovesical obstruction, and nonbilious vomiting due to pyloric atresia. Blistering of the skin developed after birth. Epidermolysis bullosa simplex was confirmed by electron microscopy of a skin biopsy specimen. We describe this patient, who had three unusual manifestations of epidermolysis bullosa. PMID- 9336806 TI - "Painful and disabling granuloma annulare": a case of Munchausen by proxy. AB - We report the unusual case of a child who from age 6 received fourteen excisions and four grafting procedures in an attempt to control "painful and disabling granuloma annulare." Although the lesions were considered disabling by the mother, they were nontender on repeated examinations. A careful investigation revealed that the mother was falsely describing symptoms necessitating surgery so she could receive public housing and other benefits for parents with a disabled child. We believe this case should alert the clinician as to the extreme measures imposed on children in Munchausen syndrome by proxy. PMID- 9336807 TI - Disseminated bacillus Calmette-Guerin infection in an HIV-infected child: a case with cutaneous lesions. AB - A boy born to a mother with unknown HIV infection was immunized with BCG in his first month of life. Seven months later axillary adenopathy developed. At the age of 10 months, 2 months after HIV infection had been diagnosed, papular skin lesions appeared all over his body. Mycobacterium bovis, BCG strain, was cultured from a lymph node and blood. Ziehl-Neelsen stain of a skin biopsy specimen showed histiocytes loaded with numerous acid-fast bacilli. The patient died 10 days later, before the infection was confirmed. This is the first reported case of disseminated BCG infection in an HIV-infected child presenting with cutaneous lesions. PMID- 9336808 TI - Ichthyosis: the skin manifestation of multiple sulfatase deficiency. AB - Juvenile sulfatidosis (Austin type) or multiple sulfatase deficiency is an extremely rare autosomal recessive disorder affecting the activity of many sulfatases: arylsulfatase A, several mucopolysaccharide sulfatases, and steroid sulfatase. Certain aspects of the clinical phenotype can be attributed mainly to a deficiency of one specific sulfatase. Most patients develop metachromatic leukodystrophy caused by arylsulfatase A deficiency, dysostosis multiplex by mucopolysaccharide sulfatase deficiency, and ichthyotic skin by steroid sulfatase deficiency. We describe a 7-year-old boy with developmental delay from 7 months of age, progressive spastic quadriparesis, and coarse facial features. By 27 months of age, an ichthyotic rash had developed on the limbs, trunk, and scalp. A skin biopsy specimen revealed hyperkeratosis with a normal granular layer. The diagnosis of multiple sulfatase deficiency was demonstrated by measuring sulfatase activities in fresh leukocytes: there were large deficiencies of arylsulfatase A and B plus reduced arylsulfatase C. The ichthyosis associated with multiple sulfatase deficiency has an autosomal recessive inheritance, is caused by steroid sulfatase deficiency, and the scaling is sometimes milder than in X-linked recessive ichthyosis. This could reflect the residual activity of steroid sulfatase in some cases. PMID- 9336809 TI - Congenital Becker's nevus with a familial association. AB - Becker's nevus is a unilateral, hyperpigmented cutaneous hamartoma usually with hypertrichosis. It occurs predominantly in boys, becoming apparent during adolescence, although several cases of congenital Becker's nevus have been reported. Rarely it may be familial and as such is transmitted in an autosomal dominant pattern. We report a 16-month-old black boy with a hyperpigmented patch on his right shoulder and upper pectoral area that extended down his arm. The patient's father has a similar lesion with hair on his left shoulder which has been present since childhood. Histology of the child's lesion was consistent with Becker's nevus. We believe this to be the first reported case of a congenital Becker's nevus with a familial association. PMID- 9336810 TI - Lymphangioma associated with Becker's nevus: a report of coincident hamartomas in a child. AB - Lymphangiomas are hamartomas which often occur during childhood. Their classification is primarily size dependent and predicts their clinical course. Larger lesions can be life threatening, but for many patients with lymphangiomas, cosmetic disfigurement is the primary concern. Treatment options are limited and have shown only variable success. Repetitive surgical excision may be necessary. We report a cystic lymphangioma of the axilla occurring in association with a Becker's nevus in an infant. Although Becker's nevi have been described in association with other hamartomas, primarily those of smooth muscle, an association with cystic lymphangioma has not been previously reported. We offer a brief discussion of possible developmental mechanisms for their coexistence. In our patient the simultaneous occurrence of a lymphangioma and a Becker's nevus appears to be a localized event, as no other developmental abnormalities are evident. PMID- 9336811 TI - Clear cell papulosis: case report and literature review. AB - Clear cell papulosis is a newly described skin disease characterized by multiple white papules. Histopathologically, diagnostic clear cells were seen among the basal cells of the epidermis. We report clear cell papulosis on the lumbar area and buttocks of a 1-year-old girl. PMID- 9336812 TI - Neonatal dermal hematopoiesis associated with diffuse neonatal hemangiomatosis. AB - This report describes a neonate with dermal hematopoiesis associated with diffuse hemangiomatosis. The cutaneous lesions consisted of multiple red papules and bluish subcutaneous nodules scattered over his body. The nodules were bluish due to the presence of hematopoietic tissue within the hemangiomas. Although neonatal dermal hematopoiesis has been described with viral infections or hematologic dyscrasias, the association with diffuse hemangiomatosis has not been previously described. PMID- 9336813 TI - Impetigo herpetiformis and Staphylococcus aureus lymphadenitis in a pregnant adolescent. AB - We report a 15-year-old primagravida female with a history of chronic plaque psoriasis who developed impetigo herpetiformis at 28 weeks gestation. Culture of a needle aspirate from a tender, enlarged cervical lymph node grew Staphylococcus aureus. The patient improved rapidly on wet dressings, topical midpotency corticosteroids, and intravenous nafcillin. The remainder of her pregnancy was uncomplicated. We speculate that both pregnancy and infection led to this pustular flare of her psoriasis. PMID- 9336814 TI - Perineal median raphe cyst. AB - Median raphe cysts occur as a developmental anomaly along the midline male perineum. A perineal median raphe cyst arising in a 3-year-old boy was successfully treated with primary excision. The possible origins and histologic features are briefly reviewed. PMID- 9336815 TI - Constipation in epidermolysis bullosa: successful treatment with a liquid fiber containing formula. AB - In epidermolysis bullosa (EB), chronic constipation, painful defecation, and fecal impaction frequently contribute to malnutrition and growth failure. Standard treatments for constipation, such as increased intake of conventional dietary fiber and fluids and/or the use of laxatives and stool softeners, are largely unsuccessful. We evaluated by questionnaire the use of a fiber-containing liquid formula (Enrich) in 20 chronically constipated children with dystrophic EB. All derived substantial improvement in constipation when taking 250 to 750 ml Enrich per day. We recommend that such a fiber-containing food be prescribed for chronic constipation in EB. In cases of fecal impaction, this should be preceded by bowel cleansing. PMID- 9336817 TI - Thigh nodule in a young girl. PMID- 9336816 TI - Iontophoresis for anesthesia during pulsed dye laser treatment of port-wine stains. AB - Port-wine stains (PWS) are benign, congenital vascular malformations found in approximately 0.3% of newborns. PWS may be effectively treated with the flashlamp pulsed dye laser (FPDL) at 585 nm. However, laser therapy of vascular lesions often produces pain. We performed a prospective double-blind, placebo-controlled evaluation of the iontophoresis of lidocaine 5% with epinephrine 1:50,000 and mepivacaine 2% with epinephrine 1:50,000. Thirty-six patients with facial PWS were included in the study; 13 of them were treated with lidocaine 5% with epinephrine, another 13 were treated with mepivacaine 2% with epinephrine, and the other 13 were treated with preservative-free 0.9% NaCl. The pain was graded by the patient on a visual analog scale from 0 to 10, comparing the iontophoretically treated area with an adjacent area treated without anesthesia. Pain evaluation by patients demonstrated a significant decrease in the pain of pulsed dye laser impulses using the iontophoresis of lidocaine 5% with epinephrine. No change in the efficacy of pulsed dye laser treatment of PWS or important side effects were observed in our patients. Iontophoresis of lidocaine 5% with epinephrine is a safe and effective method of local anesthesia for pulsed dye laser and it is more effective than the iontophoresis of mepivacaine 2% with epinephrine. PMID- 9336818 TI - What syndrome is this? Ankyloblepharon-ectodermal defects--cleft lip and palate (Hay-Wells) syndrome. PMID- 9336819 TI - Pyloric atresia and epidermolysis bullosa. PMID- 9336820 TI - Treatment resistant head lice: alternative therapeutic approaches. PMID- 9336821 TI - Abdominal Lichen aureus in a child. PMID- 9336822 TI - Dyskeratosis congenita: a case with early onset. PMID- 9336829 TI - Sequence analysis of the AAA protein family. AB - The AAA protein family, a recently recognized group of Walker-type ATPases, has been subjected to an extensive sequence analysis. Multiple sequence alignments revealed the existence of a region of sequence similarity, the so-called AAA cassette. The borders of this cassette were localized and within it, three boxes of a high degree of conservation were identified. Two of these boxes could be assigned to substantial parts of the ATP binding site (namely, to Walker motifs A and B); the third may be a portion of the catalytic center. Phylogenetic trees were calculated to obtain insights into the evolutionary history of the family. Subfamilies with varying degrees of intra-relatedness could be discriminated; these relationships are also supported by analysis of sequences outside the canonical AAA boxes: within the cassette are regions that are strongly conserved within each subfamily, whereas little or even no similarity between different subfamilies can be observed. These regions are well suited to define fingerprints for subfamilies. A secondary structure prediction utilizing all available sequence information was performed and the result was fitted to the general 3D structure of a Walker A/GTPase. The agreement was unexpectedly high and strongly supports the conclusion that the AAA family belongs to the Walker superfamily of A/GTPases. PMID- 9336830 TI - Calcium binding to tandem repeats of EGF-like modules. Expression and characterization of the EGF-like modules of human Notch-1 implicated in receptor ligand interactions. AB - The Ca(2+)-binding epidermal growth factor (cbEGF)-like module is a structural component of numerous diverse proteins and occurs almost exclusively within repeated motifs. Notch-1, a fundamental receptor for cell fate decisions, contains 36 extracellular EGF modules in tandem, of which 21 are potentially Ca(2+)-binding. We report the Ca(2+)-binding properties of EGF11-12 and EGF10-13 from human Notch-1 (hNEGF11-12 and hNEGF10-13), modules previously shown to support Ca(2+)-dependent interactions with the ligands Delta and Serrate. Ca2+ titrations in the presence of chromophoric chelators, 5,5'-Br2BAPTA and 5-NBAPTA, gave two binding constants for hNEGF11-12, Kd1 = 3.4 x 10(-5) M and Kd2 > 2.5 x 10(-4) M. The high-affinity site was found to be localized to hNEGF12. Titration of hNEGF10-13 gave three binding constants, Kd1 = 3.1 x 10(-6) M, Kd2 = 1.6 x 10( 4) M, and Kd3 > 2.5 x 10(-4) M, demonstrating that assembly of EGF modules in tandem can increase Ca2+ affinity. The highest affinity sites in hNEGF11-12 and hNEGF10-13 had 10 to 100-fold higher affinity than reported for EGF32-33 and EGF25-31, respectively, from fibrillin-1, a connective tissue protein with 43 cbEGF modules. A model of hNEGF11-12 based on fibrillin-1 EGF32-33 demonstrates electronegative potential that could contribute to the higher affinity of the Ca(2+)-binding site in hNEGF12. These data demonstrate that the Ca2+ affinity of cbEGF repeats can be highly variable among different classes of cbEGF containing proteins. PMID- 9336832 TI - Unusual conformation of nicotinamide adenine dinucleotide (NAD) bound to diphtheria toxin: a comparison with NAD bound to the oxidoreductase enzymes. AB - The conformation of NAD bound to diphtheria toxin (DT), an ADP-ribosylating enzyme, has been compared to the conformations of NAD(P) bound to 23 distinct NAD(P)-binding oxidoreductase enzymes, whose structures are available in the Brookhaven Protein Data Bank. For the oxidoreductase enzymes, NAD(P) functions as a cofactor in electron transfer, whereas for DT, NAD is a labile substrate in which the N-glycosidic bond between the nicotinamide ring and the N-ribose is cleaved. All NAD(P) conformations were compared by (1) visual inspection of superimposed molecules, (2) RMSD of atomic positions, (3) principal component analysis, and (4) analysis of torsion angles and other conformational parameters. Whereas the majority of oxidoreductase-bound NAD(P) conformations are found to be similar, the conformation of NAD bound to DT is found to be unusual. Distinctive features of the conformation of NAD bound to DT that may be relevant to DT's function as an ADP-ribosylating enzyme include (1) an unusually short distance between the PN and N1N atoms, reflecting a highly folded conformation for the nicotinamide mononucleotide (NMN) portion of NAD, and (2) a torsion angle chi N approximately 0 degree about the scissile N-glycosidic bond, placing the nicotinamide ring outside of the preferred anti and syn orientations. In NAD bound to DT, the highly folded NMN conformation and torsion angle chi N approximately 0 degree could contribute to catalysis, possibly by orienting the C1'N atom of NAD for nucleophilic attack, or by placing strain on the N glycosidic bond, which is cleaved by DT. The unusual overall conformation of NAD bound to DT is likely to reflect the structure of DT, which is unusual among NAD(P)-binding enzymes. In DT, the NAD binding site is formed at the junction of two antiparallel beta-sheets. In contrast, although the 24 oxidoreductase enzymes belong to at least six different structural classes, almost all of them bind NAD(P) at the C-terminal end of a parallel beta-sheet. The structural alignments and principal component analysis show that enzymes of the same structural class bind to particularly similar conformations of NAD(P), with few exceptions. The conformation of NAD bound to DT superimposes closely with that of an NAD analogue bound to Pseudomonas exotoxin A, an ADP-ribosylating toxin that is structurally homologous to DT. This suggests that all of the ADP-ribosylating enzymes that are structurally homologous to DT and ETA will bind a highly similar conformation of NAD. PMID- 9336831 TI - Ideal architecture of residue packing and its observation in protein structures. AB - A simple model of sphere packing has been investigated as an ideal model for long range interactions for the packing of non-bonded residues in protein structures. By superposing all residues, the geometry of packing around a central residue is investigated. It is found that all residues conform almost perfectly to this lattice model for sphere packing when a radius of 6.5 A is used to define non bonded (virtual) interacting residues. Side-chain positions with respect to sequential backbone segments are relatively regular as well. This lattice can readily be used in conformation simulations to reduce the conformational space. PMID- 9336833 TI - The structures of thymidine kinase from herpes simplex virus type 1 in complex with substrates and a substrate analogue. AB - Thymidine kinase from Herpes simplex virus type 1 (TK) was crystallized in an N terminally truncated but fully active form. The structures of TK complexed with ADP at the ATP-site and deoxythymidine-5'-monophosphate (dTMP), deoxythymidine (dT), or idoxuridine-5'-phosphate (5-iodo-dUMP) at the substrate-site were refined to 2.75 A, 2.8 A, and 3.0 A resolution, respectively. TK catalyzes the phosphorylation of dT resulting in an ester, and the phosphorylation of dTMP giving rise to an anhydride. The presented TK structures indicate that there are only small differences between these two modes of action. Glu83 serves as a general base in the ester reaction. Arg163 parks at an internal aspartate during ester formation and binds the alpha-phosphate of dTMP during anhydride formation. The bound deoxythymidine leaves a 35 A3 cavity at position 5 of the base and two sequestered water molecules at position 2. Cavity and water molecules reduce the substrate specificity to such an extent that TK can phosphorylate various substrate analogues useful in pharmaceutical applications. TK is structurally homologous to the well-known nucleoside monophosphate kinases but contains large additional peptide segments. PMID- 9336834 TI - Phosphorylation on histidine is accompanied by localized structural changes in the phosphocarrier protein, HPr from Bacillus subtilis. AB - The histidine-containing protein (HPr) of bacterial phosphoenolpyruvate:sugar phosphotransferase system (PTS) serves a central role in a series of phosphotransfer reactions used for the translocation of sugars across cell membranes. These studies report the high-definition solution structures of both the unphosphorylated and histidine phosphorylated (P-His) forms of HPr from Bacillus subtilis. Consistent with previous NMR studies, local conformational adjustments occur upon phosphorylation of His 15, which positions the phosphate group to serve as a hydrogen bond acceptor for the amide protons of Ala 16 and Arg 17 and to interact favorably with the alpha-helix macrodipole. However, the positively charged side chain of the highly conserved Arg 17 does not appear to interact directly with phospho-His 15, suggesting that Arg 17 plays a role in the recognition of other PTS enzymes or in phosphotransfer reactions directly. Unlike the results reported for Escherichia coli P-His HPr (Van Nuland NA, Boelens R, Scheek RM, Robillard GT, 1995, J Mol Biol 246:180-193), our data indicate that phosphorylation of His 15 is not accompanied by adoption of unfavorable backbone conformations for active site residues in B. subtilis P-Ser HPr. PMID- 9336836 TI - Structural heterogeneity of the various forms of apomyoglobin: implications for protein folding. AB - Temperature-induced denaturation transitions of different structural forms of apomyoglobin were studied monitoring intrinsic tryptophan fluorescence. It was found that the tryptophans are effectively screened from solvent both in native and acid forms throughout most of the temperature range tested. Thus, the tryptophans' surrounding do not show a considerable change in structure where major protein conformational transitions have been found in apomyoglobin using other techniques. At high temperatures and under strong destabilizing conditions, the tryptophans' fluorescence parameters show sigmoidal thermal denaturation. These results, combined with previous studies, show that the structure of this protein is heterogeneous, including native-like (tightly packed) and molten globule-like substructures that exhibit conformation (denaturation) transitions under different conditions of pH and temperature (and denaturants). The results suggest that the folding of this protein proceeds via two "nucleation" events whereby native-like contacts are formed. One of these events, which involves AGH "core" formation, appears to occur very early in the folding process, even before significant hydrophobic collapse in the rest of the protein molecule. From the current studies and other results, a rather detailed picture of the folding of myoglobin is presented, on the level of specific structures and their thermodynamical properties as well as formation kinetics. PMID- 9336835 TI - Identification and characterization of glycosylation sites in human serum clusterin. AB - Clusterin is a ubiquitous, heterodimeric glycoprotein with multiple possible functions that are likely influenced by glycosylation. Identification of oligosaccharide attachment sites and structural characterization of oligosaccharides in human serum clusterin has been performed by mass spectrometry and Edman degradation. Matrix-assisted laser desorption ionization mass spectrometry revealed two molecular weight species of holoclusterin (58,505 +/- 250 and 63,507 +/- 200). Mass spectrometry also revealed molecular heterogeneity associated with both the alpha and beta subunits of clusterin, consistent with the presence of multiple glycoforms. The data indicate that clusterin contains 17 27% carbohydrate by weight, the alpha subunit contains 0-30% carbohydrate and the beta subunit contains 27-30% carbohydrate. Liquid chromatography electrospray mass spectrometry with stepped collision energy scanning was used to selectively identify and preparatively fractionate tryptic glycopeptides. Edman sequence analysis was then used to confirm the identities of the glycopeptides and to define the attachment sites within each peptide. A total of six N-linked glycosylation sites were identified, three in the alpha subunit (alpha 64N, alpha 81N, alpha 123N) and three in the beta subunit (beta 64N, beta 127N, and beta 147N). Seven different possible types of oligosaccharide structures were identified by mass including: a monosialobiantennary structure, bisialobiantennary structures without or with one fucose, trisialotriantennary structures without or with one fucose, and possibly a trisialotriantennary structure with two fucose and/or a tetrasialotriantennary structure. Site beta 64N exhibited the least glycosylation diversity, with two detected types of oligosaccharides, and site beta 147N exhibited the greatest diversity, with five or six detected types of oligosaccharides. Overall, the most abundant glycoforms detected were bisialobiantennary without fucose and the least abundant were monosialobiantennary, trisialotriantennary with two fucose and/or tetrasialotriantennary. Clusterin peptides accounting for 99% of the primary structure were identified from analysis of the isolated alpha and beta subunits, including all Ser- and Thr-containing peptides. No evidence was found for the presence of O-linked or sulfated oligosaccharides. The results provide a molecular basis for developing a better understanding of clusterin structure function relationships and the role clusterin glycosylation plays in physiological function. PMID- 9336837 TI - Evaluating the energetics of empty cavities and internal mutations in proteins. AB - The energetics of cavity formation in proteins is evaluated with two different approaches and results are analyzed and compared to experimental data. In the first approach, free energy of cavity formation is extracted by RMS fitting from the distribution of numbers of cavities, N, with different volumes, Vcav, in 80 high-resolution protein structures. It is assumed that the distribution of number of cavities according to their volume follows the Boltzmann law, N(Vcav) = exp [( a.Vcav-b)/kT], or its simplified form. Specific energy cost of cavity formation, a, extracted by RMS fitting from these distributions is compared to a values extracted from experimental free energies of cavity formation in T4 lysozyme fitted to similar expressions. It is found that fitting of both sets of data leads to similar magnitudes and uncertainties in the calculated free energy values. It is shown that Boltzmann-like distribution of cavities can be derived for a simple model of an equilibrium interconversion between mutants in an extracellular system. We, however, suggest that a partitioning into cavity dependent and cavity-independent terms may lose meaning when one attempts to describe mutation effects on protein stability in terms of specific free energy contributions. As an alternative approach, a direct molecular mechanics evaluation is attempted of T4 lysozyme destabilization by five single cavity creating mutations. The calculations are based on the approach used in calculations of the energetics of packing defects in crystals. For all mutations calculated destabilizations agree with the corresponding experimental values within +/-0.6 kcal/mol. A computational relaxation of the mutant was most difficult to achieve for the mutation producing the smallest cavity. However, calculations do not always reproduce crystallographically observed contraction/expansion of cavities. It is suggested that this may be related to usually observed large RMS differences (> 1 A) between crystallographic and energy-minimized protein structures, and thus correct energetics might be easier to calculate than the correct geometry. PMID- 9336838 TI - Dissection of the gene of the bifunctional PGK-TIM fusion protein from the hyperthermophilic bacterium Thermotoga maritima: design and characterization of the separate triosephosphate isomerase. AB - Triosephosphate isomerase (TIM), from the hyperthermophilic bacterium Thermotoga maritima, has been shown to be covalently linked to phosphoglycerate kinase (PGK) forming a bifunctional fusion protein with TIM as the C-terminal portion of the subunits of the tetrameric protein (Schurig et al., EMBO J 14:442-451, 1995). To study the effect of the anomalous state of association on the structure, stability, and function of Thermotoga TIM, the isolated enzyme was cloned and expressed in Escherichia coli, and compared with its wild-type structure in the PGK-TIM fusion protein. After introducing a start codon at the beginning of the tpi open reading frame, the gene was expressed in E.c.BL21(DE3)/ pNBTIM. The nucleotide sequence was confirmed and the protein purified as a functional dimer of 56.5 kDa molecular mass. Spectral analysis, using absorption, fluorescence emission, near- and far-UV circular dichroism spectroscopy were used to compare the separated Thermotoga enzyme with its homologs from mesophiles. The catalytic properties of the enzyme at approximately 80 degrees C are similar to those of its mesophilic counterparts at their respective physiological temperatures, in accordance with the idea that under in vivo conditions enzymes occupy corresponding states. As taken from chaotropic and thermal denaturation transitions, the separated enzyme exhibits high intrinsic stability, with a half concentration of guanidinium-chloride at 3.8 M, and a denaturation half-time at 80 degrees C of 2 h. Comparing the properties of the TIM portion of the PGK-TIM fusion protein with those of the isolated recombinant TIM, it is found that the fusion of the two enzymes not only enhances the intrinsic stability of TIM but also its catalytic efficiency. PMID- 9336839 TI - Ligand binding to proteins: the binding landscape model. AB - Models of ligand binding are often based on four assumptions: (1) steric fit: that binding is determined mainly by shape complementarity; (2) native binding: that ligands mainly bind to native states; (3) locality: that ligands perturb protein structures mainly at the binding site; and (4) continuity: that small changes in ligand or protein structure lead to small changes in binding affinity. Using a generalization of the 2D HP lattice model, we study ligand binding and explore these assumptions. We first validate the model by showing that it reproduces typical binding behaviors. We observe ligand-induced denaturation, ANS and heme-like binding, and "lock-and-key" and "induced-fit" specific binding behaviors characterized by Michaelis-Menten or more cooperative types of binding isotherms. We then explore cases where the model predicts violations of the standard assumptions. For example, very different binding modes can result from two ligands of identical shape. Ligands can sometimes bind highly denatured states more tightly than native states and yet have Michaelis-Menten isotherms. Even low-population binding to denatured states can cause changes in global stability, hydrogen-exchange rates, and thermal B-factors, contrary to expectations, but in agreement with experiments. We conclude that ligand binding, similar to protein folding, may be better described in terms of energy landscapes than in terms of simpler mass-action models. PMID- 9336840 TI - Glutathione S-transferase can be used as a C-terminal, enzymatically active dimerization module for a recombinant protease inhibitor, and functionally secreted into the periplasm of Escherichia coli. AB - Glutathione S-transferase (GST) from Schistosoma japonicum, which is widely used for the production of fusion proteins in the cytoplasm of Escherichia coli, was employed as a functional fusion module that effects dimer formation of a recombinant protein and confers enzymatic reporter activity at the same time. For this purpose GST was linked via a flexible spacer to the C-terminus of the thiol protease inhibitor cystatin, whose binding properties for papain were to be studied. The fusion protein was secreted into the bacterial periplasm by means of the OmpA signal peptide to ensure formation of the two disulfide bonds in cystatin. The formation of wrong crosslinks in the oxidizing milieu was prevented by replacing three of the four exposed cysteine residues in GST. Using the tetracycline promoter for tightly controlled gene expression the soluble fusion protein could be isolated from the periplasmic protein fraction. Purification to homogeneity was achieved in one step by means of an affinity column with glutathione agarose. Alternatively, the protein was isolated via streptavidin affinity chromatography after the Strep-tag had been appended to its C terminus. The GST moiety of the fusion protein was enzymatically active and the kinetic parameters were determined using glutathione and 1-chloro-2,4-dinitrobenzene as substrates. Furthermore, strong binding activity for papain was detected in an ELISA. The signal with the cystatin-GST fusion protein was much higher than with cystatin itself, demonstrating an avidity effect due to the dimer formation of GST. The quaternary structure was further confirmed by chemical crosslinking, which resulted in a specific reaction product with twice the molecular size. Thus, engineered GST is suitable as a moderately sized, secretion-competent fusion partner that can confer bivalency to a protein of interest and promote detection of binding interactions even in cases of low affinity. PMID- 9336841 TI - Evidence for two conformational states of thioredoxin reductase from Escherichia coli: use of intrinsic and extrinsic quenchers of flavin fluorescence as probes to observe domain rotation. AB - Thioredoxin reductase (TrxR) from Escherichia coli consists of two globular domains connected by a two-stranded beta sheet: an FAD domain and a pyridine nucleotide binding domain. The latter domain contains the redox-active disulfide composed of Cys 135 and Cys 138. TrxR is proposed to undergo a conformational change whereby the two domains rotate 66 degrees relative to each other (Waksman G, Krishna TSR, Williams CH Jr, Kuriyan J, 1994, J Mol Biol 236:800-816), placing either redox active disulfide (FO conformation) or the NADPH binding site (FR conformation) adjacent to the flavin. This domain rotation model was investigated by using a Cys 138 Ser active-site mutant. The flavin fluorescence of this mutant is only 7% that of wild-type TrxR, presumably due to the proximity of Ser 138 to the flavin in the FO conformation. Reaction of the remaining active-site thiol, Cys 135, with phenylmercuric acetate (PMA) causes a 9.5-fold increase in fluorescence. Titration of the PMA-treated mutant with the nonreducing NADP(H) analogue, 3-aminopyridine adenine dinucleotide phosphate (AADP+), results in significant quenching of the flavin fluorescence, which demonstrates binding adjacent to the FAD, as predicted for the FR conformation. Wild-type TrxR, with or without PMA treatment, shows similar quenching by AADP+, indicating that it exists mostly in the FR conformer. These findings, along with increased EndoGluC protease susceptibility of PMA-treated enzymes, agree with the model that the FO and FR conformations are in equilibrium. PMA treatment, because of steric limitations of the phenylmercuric adduct in the FO form, forces the equilibrium to the FR conformer, where AADP+ binding can cause fluorescence quenching and conformational restriction favors proteolytic susceptibility. PMID- 9336842 TI - DSC studies of the conformational stability of barstar wild-type. AB - The temperature induced unfolding of barstar wild-type of bacillus amyloliquefaciens (90 residues) has been characterized by differential scanning microcalorimetry. The process has been found to be reversible in the pH range from 6.4 to 8.3 in the absence of oxygen. It has been clearly shown by a ratio of delta HvH/delta Hcal near 1 that denaturation follows a two-state mechanism. For comparison, the C82A mutant was also studied. This mutant exhibits similar reversibility, but has a slightly lower transition temperature. The transition enthalpy of barstar wt (303 kJ mol-1) exceeds that of the C82A mutant (276 kJ mol 1) by approximately 10%. The heat capacity changes show a similar difference, delta Cp being 5.3 +/- 1 kJ mol-1 K-1 for the wild-type and 3.6 +/- 1 kJ mol-1 K 1 for the C82A mutant. The extrapolated stability parameters at 25 degrees C are delta G0 = 23.5 +/- 2 kJ mol-1 for barstar wt and delta G0 = 25.5 +/- 2 kJ mol-1 for the C82A mutant. PMID- 9336843 TI - Human recombinant [C22A] FK506-binding protein amide hydrogen exchange rates from mass spectrometry match and extend those from NMR. AB - Hydrogen/deuterium exchange behavior of human recombinant [C22A] FK506 binding protein (C22A FKBP) has been determined by protein fragmentation, combined with electrospray Fourier transform ion cyclotron resonance mass spectrometry (MS). After a specified period of H/D exchange in solution, C22A FKBP was digested by pepsin under slow exchange conditions (pH 2.4, 0 degree C), and then subjected to on-line HPLC/MS for deuterium analysis of each proteolytic peptide. The hydrogen exchange rate of each individual amide hydrogen was then determined independently by heteronuclear two-dimensional NMR on 15N-enriched C22A FKBP. A maximum entropy method (MEM) algorithm makes it possible to derive the distributions of hydrogen exchange rate constants from the MS-determined deuterium exchange-in curves in either the holoprotein or its proteolytic segments. The MEM-derived rate constant distributions of C22A FKBP and different segments of C22A FKBP are compared to the rate constants determined by NMR for individual amide protons. The rate constant distributions determined by both methods are consistent and complementary, thereby validating protein fragmentation/mass spectrometry as a reliable measure of hydrogen exchange in proteins. PMID- 9336844 TI - Oligomerization properties of GCN4 leucine zipper e and g position mutants. AB - Putative intersubunit electrostatic interactions between charged amino acids on the surfaces of the dimer interfaces of leucine zippers (g-e' ion pairs) have been implicated as determinants of dimerization specificity. To evaluate the importance of these ionic interactions in determining the specificity of dimer formation, we constructed a pool of > 65,000 GCN4 leucine zipper mutants in which all the e and g positions are occupied by different combinations of alanine, glutamic acid, lysine, or threonine. The oligomerization properties of these mutants were evaluated based on the phenotypes of cells expressing lambda repressor-leucine zipper fusion proteins. About 90% of the mutants do not form stable homooligomers. Surprisingly, approximately 8% of the mutant sequences have phenotypes consistent with the formation of higher-order (> dimer) oligomers, which can be classified into three types based on sequence features. The oligomerization states of mutants from two of these types were determined by characterizing purified fusion proteins. The Type I mutant behaved as a tetramer under all tested conditions, whereas the Type III mutant formed a variety of higher-order oligomers, depending on the solution conditions. Stable homodimers comprise less than 3% of the pool; several g-e' positions in these mutants could form attractive ion pairs. Putative repulsive ion pairs are not found among the homodimeric mutants. However, patterns of charged residues at the e and g positions do not seem to be sufficient to predict either homodimer or heterodimer formation among the mutants. PMID- 9336845 TI - The specificity of carboxypeptidase Y may be altered by changing the hydrophobicity of the S'1 binding pocket. AB - The S'1 binding pocket of carboxypeptidase Y is hydrophobic, spacious, and open to solvent, and the enzyme exhibits a preference for hydrophobic P'1 amino acid residues. Leu272 and Ser297, situated at the rim of the pocket, and Leu267, slightly further away, have been substituted by site-directed mutagenesis. The mutant enzymes have been characterized kinetically with respect to their P'1 substrate preferences using the substrate series FA-Ala-Xaa-OH (Xaa = Leu, Glu, Lys, or Arg) and FA-Phe-Xaa-OH (Xaa = Ala, Val, or Leu). The results reveal that hydrophobic P'1 residues bind in the vicinity of residue 272 while positively charged P'1 residues interact with Ser297. Introduction of Asp or Glu at position 267 greatly reduced the activity toward hydrophobic P'1 residues (Leu) and increased the activity two- to three-fold for the hydrolysis of substrates with Lys or Arg in P'1. Negatively charged substituents at position 272 reduced the activity toward hydrophobic P'1 residues even more, but without increasing the activity toward positively charged P'1 residues. The mutant enzyme L267D + L272D was found to have a preference for substrates with C-terminal basic amino acid residues. The opposite situation, where the positively charged Lys or Arg were introduced at one of the positions 267, 272, or 297, did not increase the rather low activity toward substrates with Glu in the P'1 position but greatly reduced the activity toward substrates with C-terminal Lys or Arg due to electrostatic repulsion. The characterized mutant enzymes exhibit various specificities, which may be useful in C-terminal amino acid sequence determinations. PMID- 9336846 TI - Conformationally driven protease-catalyzed splicing of peptide segments: V8 protease-mediated synthesis of fragments derived from thermolysin and ribonuclease A. AB - We have studied the conformation as well as V8 protease-mediated synthesis of peptide fragments, namely amino acid residues 295-316 (TC-peptide) of thermolysin and residues 1-20 (S-peptide) of ribonuclease A, to examine whether "conformational trapping" of the product can facilitate reverse proteolysis. The circular dichroism study showed cosolvent-mediated cooperative helix formation in TC-peptide with attainment of about 30-35% helicity in the presence of 40% 1 propanol and 2-propanol solutions at pH 6 and 4 degrees C. The thermal melting profiles of TC-peptide in the above cosolvents were very similar. V8 protease catalyzed the synthesis of TC-peptide from a 1:1 mixture of the non-interacting complementary fragments (TC295-302 and TC303-316) in the presence of the above cosolvents at pH 6 and 4 degrees C. In contrast, V8 protease did not catalyze the ligation of S1-9 and S10-20, although S-peptide could assume helical conformation in the presence of the cosolvent used for the semisynthetic reaction. V8 protease was able to synthesize an analog of S-peptide (SA-peptide) in which residues 10 14 were substituted (RQHMD-->VAAAK). While S-peptide exhibited helical conformation in the presence of aqueous propanol solutions, SA-peptide displayed predominantly beta-sheet conformation. SA-peptide showed enhanced resistance to proteolysis as compared with S-peptide. Thus, failure of semisynthesis of S peptide may be a consequence of high flexibility around the 9-10 peptide bond due to its proximity to the helix stop signal. The results suggest that protease mediated ligations may be achieved by design and manipulation of the conformational aspects of the product. PMID- 9336847 TI - Urea-induced conformational changes in cold- and heat-denatured states of a protein, Streptomyces subtilisin inhibitor. AB - Streptomyces subtilisin inhibitor (SSI) is known to exist in at least two distinct denatured states, cold-denatured (D') and heat-denatured (D) under acidic conditions. In the present work, we investigated the manner how increasing urea concentration from 0 to 8 M changes the polypeptide chain conformation of SSI that exists initially in the D' and D states as well as in the native state (N), in terms of the secondary structure, the tertiary structure, and the chain form, based on the results of the experiments using circular dichroism (CD), small-angle X-ray scattering (SAXS) and 1H-NMR spectroscopy. Our results indicate that the urea-induced conformational transitions of SSI under typical conditions of D' (pH 1.8, 3 degrees C) occur at least in two steps. In the urea concentration range of 0-2 M (step 1), a cooperative destruction of the tertiary structure occurs, resulting in a mildly denatured state (DU), which may still contain a little amount of secondary structures. In the concentration range of 2 4 M urea (step 2), the DU state gradually loses its residual secondary structure, and increases the radius of gyration nearly to a maximum value. At 4 M urea, the polypeptide chain is highly disordered with highly mobile side chains. Increasing the urea concentration up to 8 M probably results in the more highly denatured or alternatively the stiffer chain conformations. The conformational transition starting from the N state proceeds essentially the same way as in the above scheme in which D' is replaced with N. The conformational transition starting from the D state lacks step 1 because the D state contains no tertiary structures and is similar to the DU state. The fact that similar conformations are reached at urea concentrations above 2 M from different conformations of D', D, and N indicates that the effect of urea dominates in determining the polypeptide conformation of SSI in the denatured states rather than the pH and temperature. PMID- 9336848 TI - 1H, 13C, and 15N backbone assignment and secondary structure of the receptor binding domain of vascular endothelial growth factor. AB - Nearly complete sequence-specific 1H, 13C, and 15N resonance assignments are reported for the backbone atoms of the receptor-binding domain of vascular endothelial growth factor (VEGF), a 23-kDa homodimeric protein that is a major regulator of both normal and pathological angiogenesis. The assignment strategy relied on the use of seven 3D triple-resonance experiments [HN(CO)CA, HNCA, HNCO, (HCA)CONH, HN(COCA)HA, HN(CA)HA, and CBCA-(CO)NH] and a 3D 15N-TOCSY-HSQC experiment recorded on a 0.5 mM (12 mg/mL) sample at 500 MHz, pH 7.0, 45 degrees C. Under these conditions, 15N relaxation data show that the protein has a rotational correlation time of 15.0 ns. Despite this unusually long correlation time, assignments were obtained for 94 of the 99 residues; 8 residues lack amide 1H and 15N assignments, presumably due to rapid exchange of the amide 1H with solvent under the experimental conditions used. The secondary structure of the protein was deduced from the chemical shift indices of the 1H alpha, 13C alpha, 13C beta, and 13CO nuclei, and from analysis of backbone NOEs observed in a 3D 15N-NOESY-HSQC spectrum. Two helices and a significant amount of beta-sheet structure were identified, in general agreement with the secondary structure found in a recently determined crystal structure of a similar VEGF construct [Muller YA et al., 1997, Proc Natl Acad Sci USA 94:7192-7197]. PMID- 9336849 TI - Protein-protein crystal-packing contacts. AB - Protein-protein contacts in monomeric protein crystal structures have been analyzed and compared to the physiological protein-protein contacts in oligomerization. A number of features differentiate the crystal-packing contacts from the natural contacts occurring in multimeric proteins. The area of the protein surface patches involved in packing contacts is generally smaller and its amino acid composition is indistinguishable from that of the protein surface accessible to the solvent. The fraction of protein surface in crystal contacts is very variable and independent of the number of packing contacts. The thermal motion at the crystal packing interface and that of the protein core, even for large packing interfaces, though the tendency is to be closer to that of the core. These results suggest that protein crystallization depends on random protein-protein interactions, which have little in common with physiological protein-protein recognition processes, and that the possibility of engineering macromolecular crystallization to improve crystal quality could be widened. PMID- 9336850 TI - Identification, expression, and crystallization of the protease-resistant conserved domain of synapsin I. AB - A 35-37-kDa protease-resistant domain of synapsin Ia/ Ib, apparently produced by low levels of endogenous proteases in vapor diffusion droplets, slowly formed crystals diffracting X-rays to approximately 10 A resolution. The fragment mainly consisted of the highly conserved C domain common to the synapsin I/II family plus short N- and C-terminal flanking segments. Two constructs (SynA and SynB) of synthetic gene fragments coding for the C domain of synapsin with or without C terminal flanking sequence were expressed in Escherichia coli as fusion proteins attached to the soluble protein glutathione-S-transferase. The fusion proteins were purified by affinity chromatography. Subsequent in situ cleavage with TEV protease resulted in the release of highly pure synapsin fragments, which were further purified by ion exchange chromatography. SynA and SynB formed crystals within three days, which diffracted to better than 3 A using a conventional X-ray source and to about 2 A using a synchrotron X-ray source. SynA crystals have the symmetry of the trigonal space groups P3(1)21 or P3(2)21 and the unit cell dimensions a = b = 77.4 A, c = 188.5 A, alpha = beta = 90 degrees, gamma = 120 degrees. SynB crystals have the symmetry of the orthorhombic space group C222(1) with the unit cell dimension a = 104.6 A, b = 113.3 A, and c = 273.8 A. PMID- 9336851 TI - Cloning, expression, and crystallization of a hyperthermophilic protein that is homologous to the eukaryotic translation initiation factor, eIF5A. AB - A gene coding for a protein homologous to a translation initiation factor of eukaryotes, eIF5A, was cloned from Methanococcus jannaschii, a hyperthermophile with an optimum growth temperature of 85 degrees C. The protein was overexpressed, purified and crystallized. The crystals were obtained by vapor diffusion method with 8% PEG 4000 as precipitant and belong to space group P4(1)22 with unit cell dimensions a = b = 45.52 A and c = 155.59 A. These crystals diffract to at least 2.2 A resolution. PMID- 9336852 TI - Accurate encoding and decoding of emotional facial expressions in schizophrenia. AB - This is a study of the encoding and decoding of emotional facial expressions by people diagnosed as schizophrenic. The results of most previous investigations have shown that schizophrenics are worse than other psychiatric and normal comparison groups at adopting and recognizing facial expressions of emotion. This study is the first in which both abilities were tested within the same group of outpatient subjects. In contrast to earlier findings, the results of this study indicate that this group of schizophrenics was equally proficient, as compared with unipolar depressive and normal medical control subjects, in the encoding and decoding of facial expressions of anger, sadness, fear, happiness, disgust, and surprise. Encoding and decoding responses in all three groups were largely unrelated. Some of the potential explanatory factors for these unusual findings include the older age of this sample and the use of a rating procedure in the decoding task that is more similar to the nature of decoding decisions made in social situations than those typically used by other investigators. The general conclusion that schizophrenics are deficient relative to comparison groups in the encoding and decoding of emotional facial expressions is not supported by these results. PMID- 9336853 TI - Spiritual recovery movements and contemporary medical care. AB - When confronted by the threat of illness, general medical and psychiatric patients may turn to treatments that have a spiritual orientation but lack empirical validation. This article examines the nature of contemporary movements that offer these treatments and their impact on medical care. A typology of spiritually oriented recovery movements is presented, including those associated with established religions, holistic medicine, or programs for self-liberation. Possible mechanisms for their behavioral and physiologic impact on health status are discussed. The psychological appeal of these treatments is analyzed in light of the way sick people may attribute meaning to illness and may then become engaged into a spiritual recovery movement, achieve a sense of self-efficacy through affiliation, and finally comply with putative "healing" practices. Although some spiritual recovery movements provide hope in the face of illness and even offer therapeutic benefits, they may also discourage patients from getting appropriate medical treatment and promote harmful regimens. Options are discussed for mental health professionals' response to the spiritual orientation of their patients and options for future research. PMID- 9336854 TI - A two-factor model of aggression. AB - This article synthesizes theoretical material from psychology research into a practical model for conceptualizing violence in psychiatric settings. Relevant research and theory are reviewed, focusing on two important behavioral models of aggressive behavior, hostile aggression and instrumental aggression. The concepts of reinforcement, anticipated rewards, specific and nonspecific stimulus-driven aggression, intermediary emotional states in aroused persons, and the aggression stimulus threshold are developed into a bimodal model applicable to the clinical management of violence. The model provides a broad framework for categorizing, understanding, and addressing aggressive behavior in clinical settings. PMID- 9336855 TI - Vamos a Traducir los MRV (let's translate the VRM): linguistic and cultural inferences drawn from translating a verbal coding system from English into Spanish. AB - Translating a verbal coding system from one language to another can yield unexpected insights into the process of communication in different cultures. This paper describes the problems and understandings we encountered as we translated a verbal response modes (VRM) taxonomy from English into Spanish. Standard translations of text (e.g., psychotherapeutic dialogue) systematically change the form of certain expressions, so supposedly equivalent expressions had different VRM codings in the two languages. Prominent examples of English forms whose translation had different codes in Spanish included tags, question forms, and "let's" expressions. Insofar as participants use such forms to convey nuances of their relationship, standard translations of counseling or psychotherapy sessions or other conversations may systematically misrepresent the relationship between the participants. The differences revealed in translating the VRM system point to subtle but important differences in the degrees of verbal directiveness and inclusion in English versus Spanish, which converge with other observations of differences in individualism and collectivism between Anglo and Hispanic cultures. PMID- 9336856 TI - Comprehensive countertransference and comprehensive treatment for the schizophrenic patient: the psychotherapeutic heart of mutative treatment. PMID- 9336857 TI - Romantic and classic visions in the therapy of psychosis: a personal perspective and evolving theory of schizophrenia. PMID- 9336858 TI - Harm reduction and decriminalization in the United States: a personal perspective. PMID- 9336859 TI - Gender differences in alcohol use and alcohol problems: mediation by social roles and gender-role attitudes. AB - This study assesses the causal place of Traditional Gender Role Attitudes (TGRA) in models for men and women, which also include social roles by explanatory variables for alcohol use and alcohol problems. Mediation of gender differences by TGRA occurs mainly in abstinence. Interaction effect is weak for alcohol consumption and frequency of "heavy drinking." The most important explanatory variables are the status factors age and education, which are mediated by TGRA in a small way. Specific aspects of alcohol-related problems are analyzed separately for the problem drinking category. Differences in results with other studies are discussed, and further study is proposed. PMID- 9336860 TI - Resilience to drinking vulnerability in women with alcoholic parents: the moderating effects of dyadic cohesion in marital communication. AB - Data from a subsample of women (N = 4,235) in two waves of the National Longitudinal Surveys of Youth (NLSY) are used to examine the relationship between parental alcoholism and alcohol use in adult life. Dyadic cohesion in marital communication (frequency of interaction and agreement on substantive issues that affect couples) is investigated as a resilience factor that could potentially mitigate adverse drinking outcomes in adult children of alcoholics (ACAs). A moderated mediation model is estimated using a Two-Stage Least Squares (2SLS) regression analysis. The results indicated that an imputed transmission of risk for drinking vulnerability in women ACAs, controlling for nonACA status, was effectively moderated by positive dyadic interaction. PMID- 9336861 TI - Alcohol use and problem drinking: prevalences in the general Rotterdam population. AB - Research was done on the distribution of abstinence, excessive drinking, alcohol related problems, and problem drinking among the general population of Rotterdam, Netherlands in 1994. Prevalences are assessed among the total population and subpopulations defined by sex, age, marital status, educational level, daily activities, and income. A general population survey was conducted among a random sample of 8,000 Dutch inhabitants of Rotterdam in the 16-69 age range. The response rate was 44% (N = 3,537). The majority of the respondents were "light" or "moderate" drinkers. Prevalences of excessive drinking, alcohol-related problems (1 or more), and problem drinking in the total population were 8, 28, and 9%, respectively. It is shown that women tend to report many alcohol-use related problems considering their relatively low consumption pattern; young men have a high prevalence of problem drinking; being single, being unemployed, and being declared unfit to work are associated with problematic drinking. The results found for socioeconomic status appear to be inconsistent. PMID- 9336862 TI - Gender and acculturation in relation to alcohol use among Hispanic (Latino) adults in two areas of the northeastern United States. AB - Telephone surveys were conducted in English or Spanish for 665 Hispanic adult residents of Long Island (New York) and Connecticut in 1992. Reported alcohol use was higher among men than women. In multivariate analyses using logistic regression, both gender and level of acculturation (positive association) were associated with drinking 1 day or more during the past month and with drinking at least 1 day per week. The association between drinking and acculturation was much stronger among women than men; thus, the gender difference in drinking declined with increasing level of acculturation. Studies are needed to delineate the processes whereby gender and acculturation influence the drinking behavior of Hispanics in different geographic areas. PMID- 9336863 TI - Gambling and problem gambling among indigenous peoples. AB - This paper compares results from studies of gambling and problem gambling among indigenous groups in New Zealand and in North Dakota. The samples for each of these studies included substantial numbers of indigenous respondents, and the methods used in these studies were similar enough to allow comparisons of Caucasian and indigenous groups from these two distinct cultures. Analysis shows that gambling involvement, gambling expenditures, and gambling-related problems are far higher among indigenous respondents than among Caucasian respondents in both New Zealand and North Dakota. These comparisons suggest that differences between indigenous peoples and Caucasians in gambling behaviors may be due to factors distinct from culture or milieu. PMID- 9336864 TI - Change in drug-using networks of injecting drug users during methadone treatment: a pilot study using snowball recruitment and intensive interviews. AB - This pilot study used snowball recruitment methods and intensive interviews to assess personal drug-using networks and HIV risk behavior of injection drug users (IDUs). Index subjects were 22 methadone maintenance patients reporting current drug injection who were interviewed about personal drug-using networks both current and prior to treatment entry. The index subjects were then asked to recruit other network members to the study. Ninety-seven network members were identified and 40 interviewed, including 18 not in treatment. Index IDUs reported fewer co-IDUs for the treatment period than the pretreatment period, suggesting a reduction in risk of exposure to HIV. The combination of snowball recruitment and intensive interview procedures constitutes a useful method for studying IDU networks. PMID- 9336865 TI - Cigarette smoking and sports participation in adolescents: a cross-sectional survey among high school students in Italy. AB - All the male students in a high school in Brescia, North Italy (about 195,000 inhabitants) in Grades 9 through 13 were given a self-administered anonymous questionnaire during school time. Among the 1,462 students who filled in a valid questionnaire, 29.1% claimed to practice one or more sports regularly (at least 4 hours/week for 9 months/year or more), 30.2% practice sports occasionally, and 40.7% no sports at all. The percentage of current smokers (at least one cigarette a month) increased from 9th grade (11.1%) to 10th (13.2%), 11th (15.2%), 12th (27.7%), and 13th (23.7%) grade. The percentage of smokers showed a steady linear increase from the lowest to the highest grade in students practicing no or occasional activity but no increase in those who regularly practice sports. Students' smoking was negatively associated with the regular practice of sports among 12th-13th grade students. PMID- 9336867 TI - The rules of drug taking: wine and poppy derivatives in the Ancient World. VIII. Lack of evidence of opium addiction. AB - The widespread therapeutic use of opium and its probable ritual use is faced with the absence of any explicit description of cases of opium dependence. It is possible that this was due to a lack of diagnostic capability. However, even the attempt to uncover cases of opium dependence by systematically analyzing the literary passages in which poppies, opium, and meconium are quoted are unsuccessful. The only two cases of suspected opium addiction that can be selected in this way are those of the Emperor Marcus Aurelius and Ovid. The most parsimonious interpretation is the lack of an hedonic use of poppy derivatives, being that this kind of use is the most frequently connected with the development of addiction. PMID- 9336866 TI - The most important unresolved issue in the addictions: conceptual chaos. AB - This article suggests that the field of addiction study and treatment remains in a state of conceptual chaos. The addictions is an emerging scientific field that lacks conceptual clarity. To develop the precision necessary for scientific advance, we must begin by developing improved definitions of substance use, abuse, dependence, and addiction. Complicating matters, psychoactive substance use is not a necessary and sufficient cause of addiction. For example, pathological gamblers experience addiction, including tolerance and withdrawal, often in the absence of any drug use. The neurobiology of subjective experience may be a more important factor in helping to explain addictive behaviors. Consequently, this article concludes that it is improper to consider drugs as the necessary precondition for addiction. Better operational definitions will advance addictions research. PMID- 9336868 TI - Opiate withdrawal outcome: the predictive ability of admission measures of motivation, self-efficacy, and lifestyle stability. AB - The ability of two models to predict the outcome of inpatient detoxification was tested. The first model related to motivation and self-efficacy for undergoing the withdrawal, and was found to be weakly predictive of premature discharges. The second model related to the stability of subjects' lifestyles prior to admission, and strongly predicted whether or not subjects would still be in receipt of withdrawal medication at the time of their discharge. It was concluded that further research was required as to the relative usefulness of cognitive/affective versus behavioral indicators of motivation for withdrawal. PMID- 9336869 TI - Patient perceptions of psychological and physiological withdrawal symptoms and positive factors associated with gradual withdrawal from methadone maintenance treatment: a prospective study. AB - Psychological and physiological withdrawal symptoms and some positive factors were studied in 10 methadone maintenance treatment patients during methadone dose reduction. The subjective ratings were made during a period of 10 days around each reduction occasion, 3 days before dose reduction and 7 days after (i.e., within the periods). To permit comparisons of the subjects' ratings between earlier and later stages of the dose reduction process, a division has been made between the first half and the second half of the total reduction occasions (i.e., between the periods). Three of the patients completed the dose reduction, while the others interrupted their withdrawal attempts. The results show that the aggregate psychological symptoms were rated low, but that, as expected, they increased significantly from the first to the second half of the dose reduction. A significant increase of the psychological symptoms also occurred from the days before each reduction to the days after. The aggregate physiological symptoms were rated very low. A significant increase in rated withdrawal intensity is found within the reduction occasions. There were no significant changes with regard to the aggregate positive factors, either within or between the reduction occasions. PMID- 9336870 TI - Hemostatic risk factors in arterial thrombosis and atherosclerosis: the thrombin fibrin and platelet-vWF axis. PMID- 9336871 TI - The in vitro and in vivo pharmacological profiles of a platelet glycoprotein IIb/IIIa antagonist, NSL-9403. AB - The in vitro and in vivo pharmacological profiles of NSL-9403 [orotyl serylarginyl-glycyl-asparatyl-tryptophane], a platelet glycoprotein IIb/IIIa (GpIIb/IIIa) antagonist, has been studied. NSL-9403 inhibited platelet aggregation of human platelet-rich plasma (PRP) with IC50 values of 4.3 +/- 0.4 microM (collagen) and 1.8 +/- 0.3 microM (ADP), which was about 100 times more potent than RGDS. It also inhibited the binding of fibrinogen to activated platelets. Ex vivo collagen and ADP-induced platelet aggregation in a guinea pig was inhibited after a bolus intravenous administration of NSL-9403 at 1.25 mg/kg and above. NSL-9403 had an anti-thrombotic effect in in vivo thrombosis models. In a platelet agonist-induced pulmonary embolic sudden death model, where a bolus injection of collagen and epinephrine induced sudden death in mice, intravenous administration of NSL-9403 before an injection of collagen and epinephrine inhibited this platelet-agonist induced death in a dose dependent manner. In an arterio-venous shunt, infusion of NSL-9403 at 3 mg/kg/hour prevented an increase in circulation pressure due to thrombus formation in the shunt circuit and platelet loss. Infusion of NSL-9403 at 1 to 10 mg/kg/hour produced a complete inhibition of platelet-dependent arterial thrombosis in a dog femoral arterial thrombosis model. Thus NSL-9403 is a potent inhibitor or platelet aggregation in vitro and a potent anti-thrombotic agent in vivo with a relatively short duration of action. PMID- 9336872 TI - Hyperalphalipoproteinemia and prostaglandin I2 stability. AB - PGI2 is a powerful regulator of thromboresistance modulating the local platelet/vessel wall interaction. Beside the amount synthesised the availability of the biologically active compound depends on its half-life at the site of action. Plasmatic half-life of PGI2 is extremely shortened during severe infections, but also in acute myocardial infarction with extremely lowered levels of HDL-c and apoAI, the latter being described as a potential PGI2-stabilising factor. These conditions are characterised by an enhanced thrombophilic risk. This study investigated for the first time whether high levels of HDL-c (mean: 95 +/- 13 mg/dl) and apoAI (mean: 179 +/- 13 mg/dl) which have been shown epidemiologically to protect against coronary heart disease in turn might be associated with an increase in PGI2 half-life. Results were obtained from 31 healthy subjects with hyperalpha-LP as compared with 10 controls. The biological half-life of PGI2 (hyperalpha-LP: mean: 915 +/- 118 sec vs. controls: 714 +/- 70 sec; p = 0.001) was positively related to HDL-c (r = 0.8795, p < 0.001) and apoAI levels (r = 0.8025, p < 0.001). The partial correlation coefficient correcting for the association between HDL-c and apoAI levels was also significant (PGI2 to HDL-c: r = 0.6000, p < 0.001). These results suggest that the antiatherosclerotic properties of HDL might be at least partly due to an increase in PGI2 half-life. PMID- 9336873 TI - Characteristics of protein kinase C-independent exocytosis in human platelets. AB - We evaluated the characteristics of the protein kinase C (PKC)-independent mechanism for ATP release in platelet-rich plasma. When ADP (10 microM) and U46619 (1 microM) were both added as agonists, a significant release was observed immediately after stimulation. The PKC inhibitor, Ro-31-7549 (10 microM), or a cyclooxygenase inhibitor, aspirin (400 microM) or indomethacin (20 microM), partially inhibited ATP release with little effect on platelet aggregation. The ATP release observed in the presence of Ro-31-7549 was abolished by a cyclooxygenase inhibitor or by preventing aggregation without stirring. In the nonstirred condition, thromboxane B2 formation was reduced by 93%. When sodium arachidonate (1 mM) rather than U46619 was used with ADP, ATP release in the presence of Ro-31-7549 was abolished by stopping the stirring with no effect on thromboxane B2 formation. In contrast, ADP/U46619-induced ATP release observed in the presence of aspirin was only partially inhibited when the stirring was stopped. This release was also inhibited dose-dependently by Ro-31-7549 at concentrations between 1 and 10 microM. These results suggest that PKC independent ATP-release in this system requires aggregation and is inhibited by a cyclooxygenase inhibitor, while PKC-dependent exocytosis is insensitive to aggregation and a cyclooxygenase inhibitor. PMID- 9336874 TI - Factor V Leiden: detection in whole blood by ASA PCR using an additional mismatch in antepenultimate position. AB - Factor V Leiden mutation was initially detected in thrombophilic patients and relatives by PCR RFLP (Restriction Fragment Length Polymorphism) according to Bertina (1). This technique presents some drawbacks and the current trend is to simplify the diagnosis. We describe a technique of Allele Specific Amplification (ASA) which is optimized in terms of reliability: an additional mismatch in antepenultimate position enables to obtain the same specificity as PCR RFLP. Furthermore, coamplification of internal control warrants an optimal sensitivity. All the PCR have been simplified: the DNA extraction improvement allows to analyse the genotype with only a few microliters of whole blood whatever the anticoagulant and the procedure of preservation (freezing, dried blood spots, storage at +4 degrees C for several days). This technique saves time. Moreover, full automation of the ASA technique may be shortened thanks to the lack of extraction and the positive/negative reading of the PCR signal. PMID- 9336875 TI - Hypercholesterolemia as a risk factor for deep-vein thrombosis. AB - Our retrospective study has shown that hyperlipidemia is a novel etiologic factor in deep-vein thrombosis (DVT) and that most of the idiopathic DVT patients were hyperlipidemic (Thrombosis Research 79, 147-151, 1995). The aim of our current study is to analyze the interrelationship between hyperlipidemia and DVT by means of a case-control study. A series of lipid parameters were analyzed using serum from 109 patients with deep vein thrombosis (DVT). One hundred nine age- and sex matched subjects served as controls. Diagnosis of hyperlipidemia was made if the serum cholesterol level was above 220 mg/dL or if the triglyceride level was above 150 mg/dL. Among several types of hyperlipidemia examined, the risk factor associated with the highest estimated odds ratio was carriage of hypercholesterolemia associated with hypertriglyceridemia (odds ratio 5.1) followed in order by hypercholesterolemia without hypertriglyceridemia (odds ratio 2.6) and hypertriglyceridemia without hypercholesterolemia (odds ratio 0.9). These findings support the hypothesis that hypercholesterolemia plays an important role in the pathogenesis of DVT. PMID- 9336877 TI - Ionizing radiation increases concentration of plasma von Willebrand factor in Cebus Apella Paraguayanus monkeys. PMID- 9336876 TI - Amounts of tPA and PAI-1 in the euglobulin fraction obtained at different pH: their relation to the euglobulin clot lysis time. PMID- 9336878 TI - Spontaneous programmed cell death (PCD) process of lymphocytes of FIV-infected cats: cellular targets and modulation. AB - Unstimulated lymphocytes from FIV-infected cats undergo spontaneous apoptosis in vitro as indicated by internucleosomal DNA fragmentation and hypodiploid DNA content of nuclei. Unlike what is reported in HIV-infected individuals, we observed that cell death of cat lymphocytes was inhibited by activation. Spontaneous apoptosis was reduced by the addition of cat serum and after activation by phorbol ester (PMA), superantigens (SEB, SEA), and to a lesser extent by mitogens such as concanavalin A and pokeweed mitogen. In contrast, apoptosis of lymphocytes from FIV-infected, but not from control cats was increased in the presence of calcium ionophore (ionomycin). Analysis of the phenotype of cells undergoing apoptosis revealed that cell death is not restricted to a cell subpopulation but involved all lymphocyte subsets. These data suggest that the mature lymphocytes of FIV-infected cats appear programmed to die by apoptosis unless rescued by specific agents, such as protein kinase C activators or mitogens. PMID- 9336879 TI - Aspects of the humoral immune response to Staphylococcus intermedius in dogs with superficial pyoderma, deep pyoderma and anal furunculosis. AB - Bacterial infection (pyoderma) of the canine skin is largely caused by Staphylococcus intermedius and may be a superficial or deep infection. Pyoderma may be a primary, idiopathic disease or secondary to a range of other dermatological disorders. In this study, the serum concentrations of IgG, IgA, antistaphylococcal IgG and antistaphylococcal IgA were measured by ELISA in normal dogs (n = 22), dogs with idiopathic deep pyoderma (n = 22), atopic dermatitis and superficial pyoderma (n = 24), atopic dermatitis without pyoderma (n = 25), flea bite dermatitis with superficial pyoderma (n = 8), pustular demodicosis (n = 8) and German shepherd dogs with anal furunculosis (n = 28). The serum IgG was significantly increased in dogs with atopy and superficial pyoderma (p < 0.001), and lower than normal in dogs with idiopathic deep pyoderma (p < 0.015). The concentration of serum IgA was significantly lower than normal in dogs with atopy uncomplicated by pyoderma (p < 0.015). The concentration of antistaphylococcal IgG in all clinical sera was significantly elevated (p < 0.001) when compared to normal dogs but concentrations of antistaphylococcal IgA were no greater than in normal dogs. Western blotting analysis for determination of the specificity of serum IgG antistaphylococcal antibody revealed that there were nine major epitopes. Discriminant analysis demonstrated that particular combinations of these epitopes were recognised more frequently by sera from dogs in different clinical groups. PMID- 9336880 TI - Comparison of in vitro function of neutrophils from cattle deficient in plasma factor XI activity and from normal animals. AB - Cattle, homozygous for the genetic disorder of factor XI (FXI) deficiency, exhibit less than 2% of normal plasma FXI activity, display an increased bleeding tendency and are more prone to infectious diseases. FXI is one of the protein components of the contact activation system of coagulation that assembles on the surface of circulating neutrophils. Because of the central role of neutrophils in inflammation, the in vitro responses of neutrophils from normal and FXI deficient cattle were compared. Neutrophil degranulation was evaluated by measuring the release of myeloperoxidase and alkaline phosphatase, and the respiratory burst was evaluated by determining superoxide anion production. Neutrophils from FXI deficient animals exhibited a significant increase (P < 0.05) in the spontaneous release of granule contents compared to the cells from normal cattle. Following stimulation with C5a complement derived from normal serum, the neutrophils from the FXI deficient animals exhibited a greater increase (P < 0.05) in both alkaline phosphatase release and superoxide production. In these neutrophils, following stimulation with C3b complement from normal serum, the relative increase in myeloperoxidase release compared to the unstimulated neutrophils was lower than that observed in the neutrophils from normal animals. There was minimal superoxide production in unactivated neutrophils from either normal or FXI deficient cattle and the response to phorbol ester stimulation was similar in both groups of animals. The C5a complement from FXI deficient serum was more effective (P < 0.05) in stimulating alkaline phosphatase release and superoxide production in normal neutrophils than the equivalent fraction from FXI deficient serum while the C3b complement from the FXI deficient serum was less effective than the normal serum fraction at inducing myeloperoxidase release from normal neutrophils. The results indicate that the differences in the in vitro neutrophil function are likely related to a variation in the function of the contact activation system on the neutrophil surface between normal and FXI deficient animals. PMID- 9336881 TI - Effects of 4-ipomeanol on bovine alveolar macrophage function. AB - The objective of this study was to determine whether 4-ipomeanol toxicosis in calves impairs alveolar macrophage functions important in pulmonary defense against infectious agents. Male Holstein calves were given either 4-ipomeanol (3 mg kg-1, i.v.) or vehicle (polyethylene glycol 400). Alveolar macrophages were recovered by pulmonary lavage 3 days later, and their capacities to phagocytose and kill E. coli, migrate toward zymosan-activated immune bovine serum, and produce interferon and interleukin-1 activity were evaluated in vitro. Alveolar macrophages recovered from 4-ipomeanol-treated calves had over a 70% decrease (p < 0.01) in chemotactic activity and over a 37% decrease (p < 0.005) in their capacity to phagocytose E. coli as compared to macrophages from control calves. Interleukin-1 activity in macrophages from 4-ipomeanol-treated calves tended to be higher than that from control calves, but the differences were not statistically significant (p = 0.06). 4-ipomeanol did not affect macrophage bactericidal activity or production of interferon. These results indicate that 4 ipomeanol suppresses select functions of alveolar macrophages in cattle that may be important in pulmonary defense against bacterial pathogens. PMID- 9336882 TI - Functional assessment of bovine monocytes isolated from peripheral blood. AB - Bovine monocytes were isolated from the peripheral blood of cattle by adherence of peripheral blood mononuclear cells to plastic. Three in vitro methods were modified to evaluate bovine monocyte function. These methods were: (1) ingestion of 125I-iododeoxyuridine labeled Staphylococcus aureus; (2) antibody-dependent cell-mediated cytotoxicity; (3) luminol-dependent and native chemiluminescence. Description of monocyte isolation and assay development is discussed in the text. Assays to evaluate monocyte function are useful for assessing immune status of the animal. PMID- 9336883 TI - Primary culture and expression of cytokine mRNAs by lipopolysaccharide in bovine Kupffer cells. AB - Kupffer cells are sessile tissue macrophages that have a role in liver defense against endogenous endotoxins. Because little information is available on the role of bovine Kupffer cells, we developed a primary culture method to investigate the function of bovine Kupffer cells. Kupffer cells were isolated from the caudate lobe of calf liver by perfusion with collagenase and pronase. Then, the cells were purified by gradient centrifugation followed by counterflow centrifugal elutriation. With the methods, a mean number of 1.5 x 10(6) Kupffer cells with a final viability of over 98% was obtained from 1 g of the liver. Over 95% of the isolated cells were positive for non-specific esterase activity and had surface molecule of CD68. The cultured Kupffer cells expressed mRNAs of tumor necrosis factor-alpha, interleukin (IL)-1 alpha, IL-1 beta and IL-6 by stimulation for 3 h with lipopolysaccharide. The primary culture of bovine Kupffer cells could be useful to investigate the systemic inflammatory response in bovine liver. PMID- 9336884 TI - Elevated levels of beta-hydroxybutyric acid in periparturient cows and in vitro effect on respiratory burst activity of bovine neutrophils. AB - The in vitro effect of normal (0.01 to 1 mM) and subketotic (1 to 2.5 mM) doses of butyric acid on the respiratory burst activity of bovine polymorphonuclear leucocytes (PMNL) isolated from blood was studied by luminol-enhanced, PMA (phorbol-12-myristate-13-acetate)-induced chemiluminescence (CL). The subketotic concentrations of butyric acid induced a significant inhibitory effect on CL. In a cell free assay, consisting of sonicated cells and H2O2, no changes in activity could be observed. The activity of the myeloperoxidase was not significantly altered as shown by the ortho-dianisidine-oxidation assay. Also, the production of O(2)- measured by the cytochrome c reduction assay was not affected by different doses of butyric acid. Butyric acid had no scavenging effect on hypochlorite. The reason for the inhibitory effect on CL may be a decreased production of H2O2. Indeed, luminol-enhanced CL evaluates the production of H2O2. This could not be confirmed in the other assays mentioned above, because H2O2 was added externally in these assays. In conclusion, because of this inhibitory effect on the respiratory burst activity of PMNL, the elevated blood level of butyric acid after parturition in high yielding cows may be, in part, responsible for the higher susceptibility to local and systemic infections during the postpartum period and during subclinical and clinical ketosis. PMID- 9336885 TI - Quantification of immunoglobulin in the serum and mucus of channel catfish at different ages and following infection with Edwardsiella ictaluri. AB - An enzyme-linked immunosorbent assay was developed to quantify immunoglobulin (Ig) in the serum and mucus of channel catfish (Ictalurus punctatus). The capture antibody (Ab) was a monoclonal Ab to channel catfish Ig and the detector was a polyclonal goat serum against channel catfish Ig. Detectable levels of channel catfish Ig were 0.1 microgram ml-1 and coefficients of intra- and interassay variations were 20% and 15%, respectively. Serum Ig concentration was found to increase from 0.36 +/- 0.25 to 0.49 +/- 0.26 mg ml-1 between 3 and 15 months of age. In contrast, mucus Ig concentration slightly decreased from 0.55 +/- 0.52 to 0.45 +/- 0.82 ng cm-2 of skin between 3 and 15 months of age. Mucus Ig concentrations were highest immediately caudal to gill covers and between pectoral to anal fins, at concentrations of 0.45 +/- 0.82 and 0.34 +/- 0.67 ng cm 2, respectively. The concentration of mucus Ig between anal and caudal fins and on the ventral skin between gill covers and pectoral fin was 0.18 +/- 0.42 and 0.09 +/- 0.14 ng cm-2, respectively. The kinetics of Ig production following infection with Edwardsiella ictaluri showed that serum Ig concentrations markedly increased (20.92 +/- 32.75 mg ml-1) at 13 days post-infection. The increase in mucus Ig concentrations occurred 14 days later (0.80 +/- 1.91 ng cm-2). Individual variation in both serum and mucus Ig production increased considerably during the course of infection (+/-0.23 to +/-32.75 mg ml-1 for serum Ig and +/ 0.1 to +3.41 ng cm-2 for mucus Ig). The results suggest that E. ictaluri infection causes higher Ig production than that found in non-infected fish of the same age. Ab response to E. ictaluri was detected in the serum but not in the mucus of infected fish. Our results show differences in serum and mucus Ig concentrations with age and after infection with E. ictaluri. PMID- 9336886 TI - Killing of Edwardsiella ictaluri by macrophages from channel catfish immune and susceptible to enteric septicemia of catfish. AB - The role of peritoneal macrophages in immunity to enteric septicemia of catfish (ESC) after infection with live Edwardsiella ictaluri was investigated. Channel catfish macrophage-mediated bacteriocidal activity was dependent on the macrophage:bacteria ratio. Ratios of 1:1 to 1:12 exhibited significant differences (P < or = 0.05) in killing between macrophages from immune fish when compared to killing by macrophages from susceptible fish at 2.5 h. At 5 h, macrophages from immune fish were capable of effective killing (83.3%) at a 1:24 effector:target ratio, whereas macrophages from susceptible fish killed significantly (P < or = 0.05) less (56.9%). Macrophage bacteriocidal activity was significantly greater (P < or = 0.05) in macrophages from individual immune fish (93.4%) compared to macrophages from individual susceptible fish (85.4%). The kinetics of macrophage killing showed a linear increase in bacteriocidal activity from 1 to 3 h. Opsonization with immune serum enabled macrophages from immune fish to kill bacteria more effectively (93.8 vs. 75.9%) at 2.5 h. Opsonization of E. ictaluri with immune serum significantly suppressed the killing ability of macrophages from susceptible fish (46.2%) at 2.5 h. The results suggest that macrophages from fish immune to ESC had a greater capacity to kill E. ictaluri than macrophages from susceptible fish especially when E. ictaluri were opsonized with anti-E. ictaluri antibody. PMID- 9336887 TI - Characterization of Borrelia lusitaniae sp. nov. by 16S ribosomal DNA sequence analysis. AB - We determined the complete sequence of the rrs gene from five strains of genomic species PotiB2. Both distance and parsimony methods were used to infer the evolutionary relationships of the rrs gene sequence of this genomic species in comparison with the rrs gene sequence of Borrelia valaisiana and the rrs gene sequences of Borrelia burgdorferi sensu lato species obtained from sequence databases. The phylogenetic analysis revealed that the genomic species PotiB2 strains clustered in a separate lineage, which was consistent with data from previous DNA-DNA hybridization experiments (D. Postic, M. V. Assous, P. A. D. Grimont, and G. Baranton, Int. J. Syst. Bacteriol. 44:743-752, 1994). A PCR restriction fragment length polymorphism analysis was used to identify genomic species PotiB2 and to differentiate it from B. burgdorferi sensu lato species. Moreover, signature nucleotide positions were identified for each B. burgdorferi sensu lato species. In accordance with DNA relatedness values, our findings suggest that genomic species PotiB2 can be more clearly defined and identified, and we propose that it should be referred to as a new species, Borrelia lusitaniae. The type strain is PotiB2. PMID- 9336888 TI - Genetic and phenotypic analysis of Borrelia valaisiana sp. nov. (Borrelia genomic groups VS116 and M19). AB - To clarify the taxonomic status of two recently described Borrelia genomic groups, groups VS116 and M19, three group VS116 strains and eight group M19 strains isolated from Ixodes ricinus ticks in Switzerland, The Netherlands, and the United Kingdom were characterized. PCR-restriction fragment length polymorphism (RFLP) analysis of the 5S-23S intergenic spacer amplicon, rRNA gene restriction analysis, 16S rRNA gene sequence analysis, randomly amplified polymorphic DNA (RAPD) fingerprinting, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting with monoclonal antibodies were used for genetic and phenotypic analysis. The PCR-RFLP and RAPD patterns of three group VS116 strains and eight group M19 strains were identical but differed from those of Borrelia burgdorferi sensu stricto, Borrelia garinii, Borrelia afzelii, and Borrelia japonica. DNAs from all group VS116 and M19 strains yielded three fragments (6.9, 3.2, and 1.4 kb) and four fragments (2.1, 1.2, 0.8, and 0.6 kb) after digestion with EcoRV and HindIII, respectively, hybridizing with an Escherichia coli 16S + 23S cDNA probe. The SDS-PAGE protein profiles of group VS116 and M19 strains were heterogeneous. Phylogenetic analysis of the partial 16S rRNA gene sequences showed that group VS116 and M19 spirochetes were members of a Borrelia species distinct from previously characterized members of the genus Borrelia. Based on our present study and data from previous DNA-DNA hybridizations, a new Borrelia species, Borrelia valaisiana sp.nov., in the B. burgdorferi complex, is proposed. Strain VS116 is the type strain of this new species. PMID- 9336889 TI - Nocardioides pyridinolyticus sp. nov., a pyridine-degrading bacterium isolated from the oxic zone of an oil shale column. AB - A bacterial strain which is able to degrade pyridine was previously isolated from the oxic zone of an oil shale column and described as Pimelobacter sp. strain OS4T. However, Pimelobacter species have been transferred to the genera Nocardioides and Terrabacter. Strain OS4T was identified as a member of the genus Nocardioides on the basis of chemotaxonomic analysis and phylogenetic inference based on 16S ribosomal DNA (rDNA) sequence analysis. The G+C content of strain OS4T is 72.5 mol%. The cell wall peptidoglycan contains LL-diaminopimelic acid as the diamino acid. The predominant menaquinone is MK-8(H4). The cellular fatty acid profile of strain OS4T is similar to that of the genus Nocardioides. The 16S rDNA similarity of strain OS4T with previously described Nocardioides species is 94.5% +/- 0.7%, and a phylogenetic tree based on 16S rDNA sequences revealed a distinct lineage for strain OS4T within the evolutionary radiation enclosed by the genus Nocardioides. Therefore, on the basis of our data, we propose that strain OS4T should be placed in the genus Nocardioides as a member of a new species, Nocardioides pyridinolyticus. The type strain of the new species is strain OS4 (= KCTC 0074BP). PMID- 9336890 TI - Deinococcus geothermalis sp. nov. and Deinococcus murrayi sp. nov., two extremely radiation-resistant and slightly thermophilic species from hot springs. AB - Strains of Deinococcus geothermalis sp. nov. were isolated from the hot spring and runoff at Agnano, Naples, Italy, and from the hot spring at Sao Pedro do Sul in central Portugal, while strains of Deinococcus murrayi sp. nov. were isolated from the hot springs at Sao Pedro do Sul, Sao Gemil, and Alcafache in central Portugal. The strains of D. geothermalis and D. murrayi produce orange-pigmented colonies and have an optimum growth temperature of about 45 to 50 degrees C. The type strains of the two new species are extremely gamma radiation resistant. The fatty acids of these new species are primarily branched-chain fatty acids. The two new species can be distinguished from each other by the lower pH range of D. geothermalis than of D. murrayi, by their fatty acid compositions, and by several biochemical parameters, including the ability of D. geothermalis to grow in minimal medium without yeast extract. 16S rRNA gene sequencing also showed that the isolates constitute two species and that these species are distinct from the other species of the genus Deinococcus. The type strain of D. geothermalis is AG 3a (= DSM 11300), and the type strain of D. murrayi is ALT-1b (= DSM 11303). PMID- 9336891 TI - Phylogeny of Photorhabdus and Xenorhabdus species and strains as determined by comparison of partial 16S rRNA gene sequences. AB - Partial 16S rRNA gene sequences of 16 strains of the genera Photorhabdus and Xenorhabdus were determined by direct sequencing of PCR products. Aligned sequences were subjected to phylogenetic analysis by maximum-likelihood and maximum-parsimony methods. Distance matrix and phylogenetic analysis did not separate the genera unambiguously. Taxonomic grouping of the bacteria closely paralleled taxonomic grouping of their nematode associates and their geographic origins. We found at least two well-supported taxonomic groups in Photorhabdus species, which suggests that the genus Photorhabdus is coevolving with the nematodes and may be polyspecific. PMID- 9336892 TI - Corynebacterium mucifaciens sp. nov., an unusual species from human clinical material. AB - Eight strains of a previously undescribed coryneform bacterium had been isolated from human clinical material over a 5-year period. Colonies of the unknown coryneform bacterium had an unusual appearance as they were slightly yellowish and very mucoid. Biochemical and chemotaxonomic characterization revealed that the unknown coryneform bacterium belonged to the genus Corynebacterium. It could be readily differentiated from all previously described Corynebacterium species. Electron microscopy demonstrated the production of an extracellular substance causing connecting filaments between cells as a morphological correlate to the mucoid colonies. Comparative 16S rRNA gene sequence analysis revealed that the unknown coryneform bacterium represented a new subline within the genus Corynebacterium, for which the name Corynebacterium mucifaciens sp. nov. is proposed. The type strain is CCUG 36878 (= DSM 44265 = CIP 105129). PMID- 9336893 TI - Borrelia recurrentis characterization and comparison with relapsing-fever, Lyme associated, and other Borrelia spp. AB - Borrelia recurrentis, the cause of louse-borne relapsing fever, has until recently been considered noncultivable, which has prevented characterization of this spirochete. We successfully cultivated 18 strains from patients with louse borne relapsing fever and present the initial characterization of these isolates. Electron microscopy revealed spirochetal cells with pointed ends, an average wavelength of 1.8 microns, an amplitude of 0.8 micron, and 8 to 10 periplasmic flagella. The G+C ratio was 28.4 mol%. Whole DNA-DNA hybridizations showed similarity between the isolates of B. recurrentis but not with Borrelia hermsii, Borrelia parkeri, Borrelia turicatae, or the Lyme-associated borreliae. Sequencing studies of both the flagellin and 16S RNA genes revealed that the greatest similarity was between B. recurrentis and Borrelia duttonii. Analysis of the sodium dodecyl sulfate-polycarylamide gel electrophoresis profiles of strains revealed four groups based on the position of a major protein band (one of the groups showed some heterogeneity and was subdivided into four subgroups). Pulsed field gel electrophoresis revealed five distinct patterns. PMID- 9336894 TI - Isolation and characterization of the homoacetogenic thermophilic bacterium Moorella glycerini sp. nov. AB - A thermophilic, anaerobic, spore-forming bacterium (strain JW/AS-Y6T) was isolated from a mixed sediment-water sample from a hot spring (Calcite Spring area) at Yellowstone National Park. The vegetative cells of this organism were straight rods, 0.4 to 0.6 by 3.0 to 6.5 microns. Cells occurred singly and exhibited a slight tumbling motility. They formed round refractile endospores in terminal swollen sporangia. Cells stained gram positive. The temperature range for growth at pH 6.8 was 43 to 65 degrees C, with optimum growth at 58 degrees C. The range for growth at 60 degrees C (pH60C; with the pH meter calibrated at 60 degrees C) was 5.9 to 7.8, with an optimum pH60C of 6.3 to 6.5. The substrates utilized included glycerol, glucose, fructose, mannose, galactose, xylose, lactate, glycerate, pyruvate, and yeast extract. In the presence of CO2, acetate was the only organic product from glycerol and carbohydrate fermentation. No H2 was produced during growth. The strain was not able to grow chemolithotrophically at the expense of H2-CO2; however, suspensions of cells in the exponential growth phase consumed H2. The bacterium reduced fumarate to succinate and thiosulfate to elemental sulfur. Growth was inhibited by ampicillin, chloramphenicol, erythromycin, rifampin, and tetracycline, but not by streptomycin. The G+C content of the DNA was 54.5 mol% (as determined by high-performance liquid chromatography). The 16S ribosomal DNA sequence analysis placed the isolate in the Gram type-positive Bacillus-Clostridium subphylum. On the basis of physiological properties and phylogenetic analysis we propose that the isolated strain constitutes a new species, Moorella glycerini; the type strain is JW/AS-Y6 (= DSM 11254 = ATCC 700316). PMID- 9336895 TI - Helicobacter salomonis sp. nov., a canine gastric Helicobacter sp. related to Helicobacter felis and Helicobacter bizzozeronii. AB - During a study of the prevalence and distribution of gastric helicobacters in domestic pets, a novel group of Helicobacter-like organisms were identified. These "Helicobacter group 2" strains were initially distinguished from the species Helicobacter felis and Helicobacter bizzozeronii by their cellular morphology and the type of motility exhibited. Bacterial cells were only slightly spiral, 5 to 7 microns long, and 0.8 to 1.2 microns wide and showed an unusual slow wavelike motion. Each cell had tufts of sheathed flagella at one or both ends. Phylogenetic analysis by 16S ribosomal DNA sequence comparison revealed that H. felis, H. bizzozeronii, "Gastrospirillum hominis" 2, and the new group of helicobacters formed a distinct cluster with intraspecies similarity values of more than 98%. These taxa were clearly separated from all other known Helicobacter species. Dot blot DNA-DNA hybridization studies indicated that the Helicobacter group 2 strains are genetically homogeneous and distinct from other canine and feline gastric helicobacters. Quantitative DNA-DNA hybridization experiments showed that Helicobacter group 2 strains exhibit > 90% DNA homology to each other, but < 39% homology to the phylogenetically related taxa H. felis and H. bizzozeronii. We propose the name Helicobacter salomonis for the novel Helicobacter group 2 strains. The type strain is H. salomonis Inkinen (= CCUG 37845). PMID- 9336896 TI - Description of Nocardiopsis synnemataformans sp. nov., elevation of Nocardiopsis alba subsp. prasina to Nocardiopsis prasina comb. nov., and designation of Nocardiopsis antarctica and Nocardiopsis alborubida as later subjective synonyms of Nocardiopsis dassonvillei. AB - Data from chemotaxonomic and 16S ribosomal DNA sequence analyses of an isolate obtained from the sputum of a kidney transplant patient identified the isolate as a member of the genus Nocardiopsis. DNA-DNA hybridization data, as well as physiological characteristics, indicated that the isolate represents a new species of the genus Nocardiopsis, designated Nocardiopsis synnemataformans; the type strain is strain IMMIB D-1215 (= DSM 44143). In addition, DNA-DNA hybridization data, as well as the results of biochemical tests, indicated that Nocardiopsis alborubida DSM 40465T, Nocardiopsis antarctica DSM 43884T, and Nocardiopsis dassonvillei DSM 43111T represent a single species designated N. dassonvillei. We also found that Nocardiopsis alba subsp. alba DSM 43377T and N. alba subsp. prasina DSM 43845T are genetically different and therefore propose that N. alba subsp. prasina be elevated to species status as Nocardiopsis prasina comb. nov., whose type strain is strain DSM 43845. PMID- 9336897 TI - Vibrio diabolicus sp. nov., a new polysaccharide-secreting organism isolated from a deep-sea hydrothermal vent polychaete annelid, Alvinella pompejana. AB - A deep-sea, facultatively anaerobic, heterotrophic, mesophilic new organism was isolated from the polychaete annelid Alvinella pompejana collected from a deep sea hydrothermal field in the East Pacific Rise. On the basis of phenotypic characteristics, phylogenetic analyses, and DNA-DNA relatedness, this organism was identified as a new species of the genus Vibrio, for which the name Vibrio diabolicus is proposed. In batch cultures in the presence of glucose, this organism produced an innovative exopolysaccharide. This polymer had high contents of both uronic acids and hexosamines and was similar to other polysaccharides with interesting biological activities. PMID- 9336898 TI - Rhizobium gallicum sp. nov. and Rhizobium giardinii sp. nov., from Phaseolus vulgaris nodules. AB - Thirty-one strains of two new genomic species (genomic species 1 and 2) of rhizobia isolated from root nodules of Phaseolus vulgaris and originating from various locations in France were compared with reference strains of rhizobia by performing a numerical analysis of 64 phenotypic features. Each genomic species formed a distinct phenon and was separated from the other rhizobial species. A comparison of the complete 16S rRNA gene sequences of a representative of genomic species 1 (strain R602spT) and a representative of genomic species 2 (strain H152T) with the sequences of other rhizobia and related bacteria revealed that each genomic species formed a lineage independent of the lineages formed by the previously recognized species of rhizobia. Genomic species 1 clustered with the species that include the bean-nodulating rhizobia, Rhizobium leguminosarum, Rhizobium etli, and Rhizobium tropici, and branched with unclassified rhizobial strain OK50, which was isolated from root nodules of Pterocarpus klemmei in Japan. Genomic species 2 was distantly related to all other Rhizobium species and related taxa, and the most closely related organisms were Rhizobium galegae and several Agrobacterium species. On the basis of the results of phenotypic and phylogenetic analyses and genotypic data previously published and reviewed in this paper, two new species of the genus Rhizobium, Rhizobium gallicum and Rhizobium giardinii, are proposed for genomic species 1 and 2, respectively. Each species could be divided in two subgroups on the basis of symbiotic characteristics, as shown by phenotypic (host range and nitrogen fixation effectiveness) and genotypic data. For each species, one subgroup had the same symbiotic characteristics as R. leguminosarum biovar phaseoli and R. etli biovar phaseoli. The other subgroup had a species-specific symbiotic phenotype and genotype. Therefore, we propose that each species should be subdivided into two biovars, as follows: R. gallicum biovar gallicum and R. gallicum biovar phaseoli; and R. giardinii biovar giardinii and R. giardinii biovar phaseoli. PMID- 9336899 TI - Recognition of two new species of intestinal spirochetes: Serpulina intermedia sp. nov. and Serpulina murdochii sp. nov. AB - On the basis of DNA-DNA hybridization data, nine intestinal spirochete strains were grouped into five genospecies. Three of these genospecies were previously recognized Serpulina species, Serpulina hyodysenteriae (type strain, B78), Serpulina innocens (type strain, B256), and Serpulina pilosicoli (type strain, P43/6/78; previously "Anguillina coli"). The other two genospecies were found to be new Serpulina species, for which we propose the names Serpulina intermedia sp. nov. (with type strain PWS/A) and Serpulina murdochii sp. nov. (with type strain 56-150). S. intermedia and S. murdochii cells had a typical spirochete ultrastructure with 22 to 28 periplasmic flagella per cell. Various soluble sugars were growth substrates for S. intermedia and S. murdochii. During growth in basal heart infusion broth supplemented with fetal calf serum beneath an O2-N2 (1:99) atmosphere, cells of these new species consumed oxygen and glucose and produced H2, CO2, acetate, butyrate, and ethanol. The G + C content of the DNA of S. murdochii 56-150T was 27 mol%, and the G + C content of the DNA of S. intermedia PWS/AT was 25 mol%. In addition, a restriction fragment length polymorphism-PCR assay for the detection of intestinal spirochetes was developed. The assay was based on generation and restriction endonuclease analysis (with HinfI, TaqI, Sau3A, and MboII) of a 558-bp amplicon of ribosomal DNA (rDNA) encoding 16S rRNA. The PCR amplification was specific for Serpulina species and Brachyspira aalborgi. Four restriction digest patterns were found for the five Serpulina species. HinfI restriction differentiated S. murdochii and S. innocens from the other species. Sau3A and TaqI restrictions gave unique fragment patterns for S. murdochii and S. pilosicoli, respectively. S. hyodysenteriae and S. intermedia DNAs gave the same fragment pattern regardless of the enzyme tested. B. aalborgi was differentiated from the Serpulina species by MboII digestion of the 16S rDNA amplicon. PMID- 9336900 TI - Thermotoga hypogea sp. nov., a xylanolytic, thermophilic bacterium from an oil producing well. AB - A new thermophilic, xylanolytic, strictly anaerobic, rod-shaped bacterium, strain SEBR 7054T, was isolated from an African oil-producing well. Based on the presence of an outer sheath (toga) and 16S rRNA sequence analysis data, this organism was identified as a member of the genus Thermotoga. Strain SEBR 7054T possessed lateral flagella, had a G + C content of 50 mol%, produced traces of ethanol from glucose but no lactate, and grew optimally in the presence of 0 to 0.2% NaCl at 70 degrees C. Its phenotypic and phylogenetic characteristics clearly differed from those reported for the five previously validly described Thermotoga species. Therefore, we propose that strain SEBR 7054T is a member of a new species of the genus Thermotoga, Thermotoga hypogea sp. nov. The type strain of T. hypogea is SEBR 7054 (= DSM 11164). PMID- 9336901 TI - Taxonomy of Pseudomonas strains isolated from tomato pith necrosis: emended description of Pseudomonas corrugata and proposal of three unnamed fluorescent Pseudomonas genomospecies. AB - Thirty-three fluorescent Pseudomonas strains isolated from tomato pith necrosis (FPTPN strains) and 89 Pseudomonas corrugata strains were studied by numerical taxonomy. In the dendrogram of distances, the P. corrugata strains constituted a single phenon (phenon 1), whereas 17 of the 33 FPTPN strains clustered in a separate phenon (phenon 2). The other 16 FPTPN strains were included in phena consisting of well-characterized fluorescent Pseudomonas species or were isolated phenotypes. Phena 1 and 2 were distinguished by fluorescence on King B medium, accumulation of poly-beta-hydroxybutyrate, production of levan, and assimilation of sorbitol. DNA-DNA hybridization showed that P. corrugata is a true genomic species (66 to 100% DNA relatedness) and that the FPTPN strains of phenon 2 were divided into three genomic groups. Genomic groups 1 and 2 were not distinct from each other phenotypically, and genomic group 3 could be distinguished from genomic groups 1 and 2 only on the basis of assimilation of citraconate and laevulinate. Genomic groups 1 and 2 are related to P. corrugata (40 to 55% DNA relatedness), whereas genomic group 3 is less closely related to P. corrugata (20 to 23% DNA relatedness). The lipopolysaccharide patterns on electrophoresis gels and fatty acid profiles of strains belonging to genomic group 1 through 3 are different from each other and from the lipopolysaccharide patterns and fatty acid profiles of P. corrugata. However, cross-reactions were observed between P. corrugata and the FPTPN strain genomic groups, indicating that there are common epitopes of the lipopolysaccharides. The three FPTPN strain genomic groups were not named as species but were designated Pseudomonas genomospecies FP1, FP2, and FP3. PMID- 9336902 TI - Shewanella woodyi sp. nov., an exclusively respiratory luminous bacterium isolated from the Alboran Sea. AB - Thirty-four strains of nonfermentative, respiratory, luminous bacteria were isolated from samples of squid ink and seawater from depths of 200 to 300 m in the Alboran Sea. Although these strains had a few properties similar to properties of Shewanella (Alteromonas) hanedai, they did not cluster phenotypically with any previously described bacterium. The nucleotide sequence of a 740-bp segment of luxA was not homologous with other known luxA sequences but clustered with the luxA sequences of Shewanella hanedai, Vibrio logei, Vibrio fischeri, and Photobacterium species. The 16S RNA gene from two strains was sequenced and was found to be most closely related to the S. hanedai 16S RNA gene. Based on the differences observed, we describe the new isolates as members of new species, Shewanella woodyi sp. nov. Strain ATCC 51908 (= MS32) is the type strain of this new species. PMID- 9336903 TI - Shewanella gelidimarina sp. nov. and Shewanella frigidimarina sp. nov., novel Antarctic species with the ability to produce eicosapentaenoic acid (20:5 omega 3) and grow anaerobically by dissimilatory Fe(III) reduction. AB - A polyphasic taxonomic study was performed to characterize dissimilatory iron reducing strains mostly isolated from Antarctic sea ice. The strains were isolated from samples of congelated (land-fast) sea ice, grease ice, and ice algal biomass collected from the coastal areas of the Vestfold Hills in eastern Antarctica (68 degrees S 78 degrees E). The strains were facultatively anaerobic, motile, and rod shaped, were capable of anaerobic growth either by fermentation of carbohydrates or by anaerobic respiration, and utilized a variety of electron acceptors, including nitrate, ferric compounds, and trimethylamine N-oxide. A phylogenetic analysis performed with 16S rRNA sequences showed that the isolates formed two groups representing novel lineages in the genus Shewanella. The first novel group included seawater-requiring, psychrophilic, chitinolytic strains which had DNA G + C contents of 48 mol%. The members of the second strain group were psychrotrophic and did not require seawater but could tolerate up to 9% NaCl. The strains of this group were also unable to degrade polysaccharides but could utilize a number of monosaccharides and disaccharides and had G + C contents of 40 to 43 mol%. The whole-cell-derived fatty acid profiles of the sea ice isolates were found to be similar to the profiles obtained for other Shewanella species. The omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) (20:5 omega 3) was detected in all of the sea ice isolates at levels ranging from 2 to 16% of the total fatty acids. EPA was also found at high levels in Shewanella hanedai (19 to 22%) and Shewanella benthica (16 to 18%) but was absent in Shewanella alga and Shewanella putrefaciens. On the basis of polyphasic taxonomic data, the Antarctic iron-reducing strains are placed in two new species, Shewanella frigidimarina sp. nov. (type strain, ACAM 591) and Shewanella gelidimarina sp. nov. (type strain, ACAM 456). PMID- 9336904 TI - A proposal to revive the genus Kitasatospora (Omura, Takahashi, Iwai, and Tanaka 1982). AB - We determined almost complete 16S ribosomal DNA sequences for 12 actinomycete strains which were either previously classified as Kitasatospora strains or defined as Streptomyces strains but shown to contain major amounts of meso diaminopimelic acid in their whole-cell hydrolysates. These sequences were subjected to phylogenetic analyses together with the sequences of 34 Streptomyces species. Phylogenetic trees were reconstructed by using both neighbor-joining and maximum-parsimony methods. The Kitasatospora species always formed a stable monophyletic clade. However, the genus Kitasatospora appeared to be either a sister taxon of the genus Streptomyces or a lineage that originated from within Streptomyces species, depending on the outgroup used. Phylogenetic trees were also constructed by using the sequences of the 16S-23S rRNA gene spacers. Streptomyces and Kitasatospora species were consistently recovered as two distinct clades independent of the outgroup used. On the basis of phylogenetic, chemotaxonomic, and phenotypic evidence, we propose that the genus Kitasatospora Omura et al. 1982 should be revived. PMID- 9336905 TI - Campylobacter hyoilei Alderton et al. 1995 and Campylobacter coli Veron and Chatelain 1973 are subjective synonyms. AB - The taxonomic affiliation of Campylobacter hyoilei was reevaluated by examining a variety of phenotypic and genotypic criteria. Whole-cell protein electrophoresis and a comparison of 66 phenotypic characters revealed that reference strains of C. hyoilei were indistinguishable from Campylobacter coli strains. These data were confirmed by a DNA-DNA hybridization level of 67% between the type strains of the two species. Several species-specific assays based on PCR amplification or probe hybridization further substantiated that C. coli strains and C. hyoilei strains are indistinguishable. It is therefore concluded that C. hyoilei and C. coli represent the same species and that the former name should be regarded as a junior synonym of the latter name. PMID- 9336906 TI - Phylogenetic analysis of Erwinia species based on 16S rRNA gene sequences. AB - The phylogenetic relationships of the type strains of 16 Erwinia species were investigated by performing a comparative analysis of the sequences of the 16S rRNA genes of these organisms. The sequence data were analyzed by the neighbor joining method, and each branch was supported by moderate bootstrap values. The phylogenetic tree and sequence analyses confirmed that the genus Erwinia is composed of species that exhibit considerable heterogeneity and form four clades that are intermixed with members of other genera, such as Escherichia coli, Klebsiella pneumoniae, and Serratia marcescens. Cluster I includes the type strains of Erwinia herbicola, Erwinia milletiae, Erwinia ananas, Erwinia uredovora, and Erwinia stewartii and corresponds to Dye's herbicola group. Cluster II consists of Erwinia persicinus, Erwinia rhapontici, Erwinia amylovora, and Erwinia cypripedii. Cluster III consists of Erwinia carotovora subspecies and Erwinia chrysanthemi and is characterized by the production of pectate lyases and cellulases. Erwinia salicis, Erwinia rubrifaciens, and Erwinia nigrifluens form the cluster that is most distantly related to other Erwinia species. The data from the sequence analyses are discussed in the context of biochemical and DNA DNA hybridization data. PMID- 9336907 TI - Methanogenium frigidum sp. nov., a psychrophilic, H2-using methanogen from Ace Lake, Antarctica. AB - Methanogenium frigidum sp. nov. was isolated from the perennially cold, anoxic hypolimnion of Ace Lake in the Vesfold Hills of Antarctica. The cells were psychrophilic, exhibiting most rapid growth at 15 degrees C and no growth at temperatures above 18 to 20 degrees C. The cells were irregular, nonmotile coccoids (diameter, 1.2 to 2.5 microns) that occurred singly and grew by CO2 reduction by using H2 as a reductant. Formate could replace H2, but growth was slower. Acetate, methanol, and trimethylamine were not catabolized. Cells grew with acetate as the only organic compounds in the culture medium, but growth was much faster in medium also supplemented with peptones and yeast extract. The cells were slightly halophilic; good growth occurred in medium supplemented with 350 to 600 mM Na+, but no growth occurred with 100 or 850 mM Na+. The pH range for growth was 6.5 to 7.9; no growth occurred at pH 6.0 or 8.5. Growth was slow (maximum specific growth rate, 0.24 day-1; doubling time, 2.9 days). This is the first report of a psychrophilic methanogen growing by CO2 reduction. PMID- 9336908 TI - Streptococcus hyovaginalis sp. nov. and Streptococcus thoraltensis sp. nov., from the genital tract of sows. AB - Two groups of strains isolated from sows were shown to belong to new sublines in the genus Streptococcus. Based on phenotypic and phylogenetic analyses, we propose that these bacteria should be classified as two new species, Streptococcus hyovaginalis sp. nov. and Streptococcus thoraltensis sp. nov. These two species are found in the genital tract, but the capnophilic species S. thoraltensis may also occur in the intestinal tract of pigs. The type strain of S. hyovaginalis is SHV515 (= LMG 14710), and S69 (= LMG 13593) is the type strain of S. thoraltensis. PMID- 9336909 TI - Spiroplasma lineolae sp. nov., from the horsefly Tabanus lineola (Diptera: Tabanidae). AB - Spiroplasma strain TALS-2T from the viscera of the striped horsefly, Tabanus lineola, collected in Georgia was serologically distinct from other Spiroplasma species, groups, putative groups, and subgroups. Light and electron microscopy of cells of strain TALS-2T revealed helical motile cells surrounded only by a single cytoplasmic membrane. The organism grew in M1D and SP-4 liquid media. Growth also occurred in 1% serum fraction medium and in conventional horse serum medium. Growth in liquid media was serum dependent. The strain passed through 220-nm filter pores, but was retained in filters with 100-nm pores. The optimum temperature for growth was 30 degrees C. Multiplication occurred at temperatures from 20 to 37 degrees C, with a doubling time at the optimum temperature of 5.6 h in M1D broth. Strain TALS-2T catabolized glucose but hydrolyzed neither arginine nor urea. The guanine-plus-cytosine content of the DNA was 25 +/- 1 mol%. The genome size was 1,390 kbp. Six isolates serologically similar to strain TALS-2T were obtained from the same host in coastal Georgia. Three strains closely related to strain TALS-2T were isolated from the horsefly Poeciloderas quadripunctatus in Costa Rica. Strain TALS-2T (= ATCC 51749), a representative of group XXVII, is designated the type strain of a new species, Spiroplasma lineolae (Mollicutes: Entomoplasmatales). PMID- 9336910 TI - Corynebacterium mastitidis sp. nov., isolated from milk of sheep with subclinical mastitis. AB - Fourteen strains of a hitherto unknown catalase-positive, aerobic, gram-positive coryneformlike organism were isolated from the milk of sheep with subclinical mastitis from different regions of Spain. The strains phenotypically closely resembled one another and biochemically were similar to Corynebacterium urealyticum and Corynebacterium afermentans subsp. lipophilum. The results of chemotaxonomic investigations were consistent with membership in the genus Corynebacterium, and comparative 16S rRNA gene sequencing studies showed that the unknown bacterium from sheep was indeed a member of the genus Corynebacterium. Within the genus Corynebacterium the new bacterium formed a distinct subline that exhibited > 4% sequence divergence with other species. Based on both phenotypic and phylogenetic findings, a new species, Corynebacterium mastitidis, is proposed for the organisms from mastitic sheep. The type strain of C. mastitidis is CECT 4843 (= S-8). PMID- 9336911 TI - Biodiversity of bradyrhizobia nodulating Lupinus spp. AB - The genetic structure of Bradyrhizobium isolates recovered from three Lupinus species (Lupinus campestris, Lupinus montanus, and Lupinus exaltatus) grown in Mexico was examined. Among 41 Bradyrhizobium isolates, 18 electrophoretic types (ETs) were distinguished by multilocus enzyme electrophoresis of five metabolic enzymes. The mean genetic diversity, 0.64, indicated that there was great genetic diversity in the population sampled. Most isolates (63%) fell into two closely related clusters (clusters I and II) and were the types most frequently isolated from the root nodules of L. montanus and L. campestris. ET cluster III isolates were frequent nodule occupants of L. exaltatus. The isolates also were assigned to three main groups by using Curie point pyrolysis mass spectrometry. In general, the multilocus enzyme electrophoretic data and pyrolysis mass spectrometric data agreed. We determined the 16S rRNA sequences of representative Lupinus isolates and of Bradyrhizobium japonicum USDA 6T and found that the lupine isolates were highly related to the B. japonicum type strain, although not all B. japonicum type strains (subcultures maintained in different bacterial collections) had identical small-subunit rRNA. PMID- 9336912 TI - Corynebacterium singulare sp. nov., a new species for urease-positive strains related to Corynebacterium minutissimum. AB - We studied two coryneform strains from clinical specimens. These strains had type IV and corynemycolic acids in their cell walls and also had phenotypic characteristics, such as urease activity and fermentation of glucose and sucrose but not trehalose, which did not permit assignment to any previously recognized taxon. According to DNA-DNA hybridization data, these two strains are members of the same species (level of DNA similarity, 86%). Phylogenetic analysis based on comparisons of almost complete small-subunit ribosomal DNA sequences revealed that these strains are closely related to Corynebacterium minutissimum, but DNA relatedness experiments clearly showed that they constitute a distinct new species with a level of DNA relatedness to the C. minutissimum type strain of less than 40%. This new species can be differentiated from C. minutissimum strains by its enzymatic activities and carbon source utilization, and the name Corynebacterium singulare is proposed for it. The type strain is strain IBS B52218 (= CCUG 37330), which was isolated from a semen specimen. PMID- 9336913 TI - Classification of Austrian rhizobia and the Mexican isolate FL27 obtained from Phaseolus vulgaris L. as Rhizobium gallicum. AB - The phylogenetic positions of four rhizobial strains obtained from nodules of common bean plants (Phaseolus vulgaris L.) grown in an Austrian soil and of the Mexican bean isolate FL27 are described. Analysis of the 16S rRNA genes revealed sequences almost identical to that of the Rhizobium gallicum type strain, R602sp, with a maximum of two nucleotide substitutions. Comparison of the 16S rRNA gene sequences with those from other bacteria indicated highest similarity to Rhizobium sp. strain OK-50, Rhizobium leguminosarum IAM 12609, and Rhizobium etli. DNA homology determined by DNA-DNA hybridization was high among the Austrian isolates and R602spT (45 to 90%) and ranged from 21 to 65% with FL27, but hybridization analysis revealed very low homology to the recognized common bean-nodulating species, R. leguminosarum bv. phaseoli, R. etli, and Rhizobium tropici. Ribosomal gene organization was studied by Southern hybridization with the 16S rRNA gene and temperature gradient gel electrophoresis, indicating identical organizations and the presence of three identical 16S rRNA copies in the genome of this species. The six strains investigated showed different plasmid profiles based on their geographical origins. We propose that the Austrian isolates and the Mexican strain FL27 are members of the species R. gallicum. PMID- 9336914 TI - Reexamination of yeast strains classified as Torulaspora delbrueckii (Lindner). AB - Twenty-eight yeast strains presumed to represent Torulaspora delbrueckii were analyzed by randomly amplified polymorphic DNA-PCR analysis. Four strains (HUT 7161, IFO 1138, IFO 1145, and IFO 1956) that were considerably different from the type strain were further investigated. Morphological and physiological characteristics revealed that strains HUT 7161 and IFO 1145 belong to the genus Debaryomyces rather than the genus Torulaspora, and the former strain may represent Debaryomyces hansenii. Strains IFO 1138 and IFO 1956 were classified as either Saccharomyces castellii or Saccharomyces dairensis by identification keys involving physiological tests. On the basis of analysis of the sequences of two rRNA internal spacer regions, strains IFO 1138 and IFO 1956 were closely related to S. castellii and strains HUT 7161 and IFO 1145 were outside members of the genera Torulaspora, Zygosaccharomyces, and Saccharomyces. PMID- 9336915 TI - Corynebacterium durum sp. nov., from human clinical specimens. AB - A new Corynebacterium species, Corynebacterium durum, was isolated from respiratory tract specimens of five human patients. The strains of this species exhibited similar morphologic and biochemical features that differentiated them from all recognized species. Notably, all of these strains developed irregular and strongly adherent colonies under aerobic conditions and produced acid from mannitol and galactose. The cells are long pleomorphic rods with some filaments. This species has characteristics of the genus Corynebacterium, such as 55 mol% guanine plus cytosine in the DNA and the presence of corynomycolic acids, meso diaminopimelic acid, arabinose, and galactose in the cell wall. These isolates formed a homogeneous group in which the DNA-DNA similarity values (as determined by an S1 nuclease procedure) compared with reference strain IBS G15036T (T = type strain) ranged from 71 to 100%. The analysis of the nearly complete 16S rRNA gene sequence of IBS G15036T indicated that this new species represents a distinct taxon within the genus Corynebacterium. This new species can be identified on the basis of its colony morphology, fermentation of sugars, and enzymatic activities. Strain IBS G15036 (= CCUG 37331) is the type strain of C. durum. PMID- 9336916 TI - Borrelia burgdorferi sensu stricto, a bacterial species "made in the U.S.A."? AB - Among the three main species of Borrelia burgdorferi sensu lato associated with Lyme borreliosis, B. burgdorferi sensu stricto (B. burgdorferi) is the sole species present both in North America and in Europe, where Borrelia garinii and Borrelia afzelii also occur. The greater genetic diversity together with the greater clinical polymorphism observed in the Old World suggests that this is the birthplace of the complex B. burgdorferi sensu lato. However, the genetic proximity of some North American and European B. burgdorferi strains in quite mystifying. A previous study of the whole genome (M. Foretz, D. Postic, and G. Baranton, Int. J. Syst. Bacteriol. 47:11-18, 1997) compared the diversity of North American and European B. burgdorferi strains. To further investigate the geographical origin and the migration of B. burgdorferi, we have focused on the study of the single variable and highly adaptive gene ospC. Both approaches demonstrated the greater diversity of North American strains and the close relatedness between European strains and between some isolates from the two areas. We discuss the significance of these features and suggest that they might be evidence of the anteriority of North American B. burgdorferi strains. PMID- 9336917 TI - Thermosipho melanesiensis sp. nov., a new thermophilic anaerobic bacterium belonging to the order Thermotogales, isolated from deep-sea hydrothermal vents in the southwestern Pacific Ocean. AB - A new thermophilic, anaerobic rod-shaped bacterium, strain BI429T was isolated from the gills of a deep-sea vent hydrothermal mussel, Bathymodiolus brevior, from the Lau Basin (Southwestern Pacific Ocean). Phenotypically, this isolate exhibited characteristics similar to those described for members of the order Thermotogales. This organism was identified as a member of the genus Thermosipho on the basis of the presence of the typical outer sheath-like structure (toga), its 16S rRNA sequence, and its ability to grow on carbohydrates (sucrose, starch, glucose, maltose, lactose, cellobiose, and galactose). The cells of this organism were gram negative and rod shaped and generally occurred singly or in pairs, rarely occurring as chains with a maximum of five rods. At the optimum temperature for growth (70 degrees C), optimum pH (6.5), and optimum salinity (30 g of NaCl per liter), the doubling time was 100 min. In spite of the high percentage of similarity of its 16S rRNA sequence with that of Thermosipho africanus (98.6%), the weak level of DNA-DNA reassociation with this strain (2%) and particular physiological characteristics allowed us to differentiate this new organism from the sole species of the genus Thermosipho previously described (T. africanus). On the basis of these observations, we propose that the new organism should be described as a new species, Thermosipho melanesiensis. The type strain of T. melanesiensis is BI429. PMID- 9336918 TI - Desulfobacter vibrioformis sp. nov., a sulfate reducer from a water-oil separation system. AB - A mesophilic, gram-negative, vibrio-shaped, marine, acetate-oxidizing sulfate reducer (strain B54) was isolated from a water-oil separation system on a North Sea oil platform. The optimum conditions for growth were 33 degrees C, pH 6.8 to 7.0, and concentrations of NaCl and MgCl2.6H2O of at least 1 and 0.3%, respectively. Of various organic acids tested, only acetate was used as an electron and carbon source. The presence of 2-oxoglutarate:dye oxidoreductase suggests acetate oxidation via an operative citric acid cycle. Even though growth of most Desulfobacter strains (including strain B54) did not occur on hydrogen, hydrogenase was detected at low activity. The growth yields were 4.6, 13.1, and 9.6 g of (dry weight) cells per mol of acetate oxidized with sulfate, sulfite, and thiosulfate, respectively, as electron acceptors. Strain B54 was able to fix dinitrogen. Desulforubidin and cytochromes of the c and b types were present. The G+C content of the DNA was 47 mol%. Strain B54 is most closely related to Desulfobacter latus, with a 16S rDNA sequence similarity of 98.1%. The DNA-DNA relatedness between them was 40.5%. On the basis of differences in genotypic, phenotypic, and immunological characteristics, we propose that strain B54 is a member of a new species, D. vibrioformis. It can be easily identified and distinguished from other Desulfobacter species by its large, vibrioshaped cells. PMID- 9336919 TI - Demetria terragena gen. nov., sp. nov., a new genus of actinomycetes isolated from compost soil. AB - A novel actinomycete was isolated from compost soil and was studied taxonomically and phylogenetically. Cells of this organism were gram positive, not acid fast, nonmotile, nonsporulating, irregular coccoid to short rod shaped, and microaerophilic. The cell wall peptidoglycan contained lysine and was cross linked via an L-Lys<--L-Ser<--D-Asp interpeptide bridge. The major menaquinone was MK-8(H4). The polar lipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and two unknown phospholipids. Mycolic acids were absent. The cellular fatty acid profile was complex, with large amounts of saturated and monounsaturated straight-chain acids and smaller amounts of iso and anteiso branched-chain acids. The G+C content of the DNA was 66 mol%. Comparative 16S ribosomal DNA studies revealed that strain HKI 0089T represents a novel lineage within Actinobacteria (32) distinct from all previously described genera and most closely related to members of the genera Kytococcus, Dermacoccus, and Dermatophilus of the family Dermatophilaceae. On the basis of our results, we suggest that strain HKI 0089 should be classified in a new genus and species, for which we propose the name Demetria terragena. The type strain and the only strain of the genus and species is HKI 0089 (DSM 11295). PMID- 9336920 TI - Phylogenetic analysis of the genus Desulfotomaculum: evidence for the misclassification of Desulfotomaculum guttoideum and description of Desulfotomaculum orientis as Desulfosporosinus orientis gen. nov., comb. nov. AB - Almost complete 16S ribosomal DNA (rDNA) sequences were determined for the type strains of nine species belonging to the genus Desulfotomaculum and for seven strains described as strains of this genus. The sequences were compared with previously published 16S rDNA and rRNA sequences of the type strains of the other species of the genus. The majority of the species form a phylogenetically coherent cluster within the Clostridium-Bacillus subphylum of gram-positive bacteria. The cluster consists of phylogenetically well-separated lineages containing (i) Desulfotomaculum nigrificans, Desulfotomaculum aeronauticum, and Desulfotomaculum ruminis, (ii) Desulfotomaculum geothermicum, Desulfotomaculum thermosapovorans, and Desulfotomaculum sapomandens, (iii) Desulfotomaculum kuznetsovii, Desulfotomaculum australicum, and Desulfotomaculum thermocisternum, (iv) Desulfotomaculum thermobenzoicum and Desulfotomaculum thermoacetoxidans, and (v) Desulfotomaculum acetoxidans. Some as-yet-undescribed Desulfotomaculum strains are phylogenetically well-separated from strains of the described species. Desulfotomaculum guttoideum shares extremely high 16S rDNA similarity with certain Clostridium species (e.g., Clostridium sphenoides and Clostridium celerecrescens) and is most likely a misidentified species. Desulfotomaculum orientis represents a new genus which branches most closely to the genus Desulfitobacterium. The name Desulfosporosinus orientis gen. nov., comb. nov., is proposed for this taxon. PMID- 9336921 TI - In vitro culture and phylogenetic analysis of "Candidatus Arsenophonus triatominarum," an intracellular bacterium from the triatomine bug, Triatoma infestans. AB - An intracellular symbiotic bacterium was isolated from the hemolymph of Triatoma infestans and cultured in an Aedes albopictus cell line. 16S ribosomal DNA sequence analysis revealed that the bacterium was a member of the gamma-3 subgroup of the class Proteobacteria, having 96.2% sequence identity with the most closely related bacterium, Arsenophonus nasoniae, the causative agent of the son-killer trait in the parasitoid wasp Nasonia vitripennis. These bacteria share morphological features and a common tissue distribution and transmission mode. The A. nasoniae-T. infestans symbiont branch represents a lineage of insect symbionts which may be capable of horizontal transmission between phylogenetically distant host insects. We propose that the intracellular symbiont from T. infestans be classified as "Candidatus Arsenophonus triatominarum." The bacterium found in the hemocytes of T. infestans is designated the type strain of this species. PMID- 9336922 TI - Discovery and classification of ecological diversity in the bacterial world: the role of DNA sequence data. AB - All living organisms fall into discrete clusters of closely related individuals on the basis of gene sequence similarity. Evolutionary genetic theory predicts that in the bacterial world, each sequence similarity cluster should correspond to an ecologically distinct population. Indeed, surveys of sequence diversity in protein-coding genes show that sequence clusters correspond to ecological populations. Future population surveys of protein-coding gene sequences can be expected to disclose many previously unknown ecological populations of bacteria. Sequence similarity clustering in protein-coding genes is recommended as a primary criterion for demarcating taxa. PMID- 9336923 TI - Inclusion of Aeromonas DNA hybridization group 11 in Aeromonas encheleia and extended descriptions of the species Aeromonas eucrenophila and A. encheleia. AB - The recently reported chemotaxonomic and genotypic description of two well separated subgroups (I and II) in Aeromonas eucrenophila and their affiliation to Aeromonas encheleia and the unnamed Aeromonas DNA hybridization group (HG) 11 (G. Huys, M. Altwegg, M.-L. Hanninen, M. Vancanneyt, L. Vauterin, R. Coopman, U. Torck, J. Luthy-Hottenstein, P. Janssen, and K. Kersters, Syst. Appl. Microbiol. 19:616-623, 1996) has questioned the original species descriptions of A. eucrenophila and A. encheleia. In order to elucidate the unclear taxonomic status of these taxa in the genus Aeromonas, we have further investigated a collection of 14 reference strains and 14 related isolates encompassing the taxa A. eucrenophila subgroups I and II, A. encheleia, and HG11 by DNA-DNA hybridization (on 17 of the 28 strains) and phenotypic characterization (on all 28 strains). Genotypically, the investigated strains could be grouped into two DNA hybridization groups that exhibited between-group homologies ranging from 42 to 52%. The members of DNA homology group I (DNA binding, 76 to 100%) were strains of A. eucrenophila subgroup I, including the type strain LMG 3774, and two A. eucrenophila-like isolates, leading to the conclusion that these strains should be considered true representatives of the species A. eucrenophila. The strains of A. eucrenophila subgroup II, HG11, and A. encheleia, on the other hand, were closely joined in DNA homology group II (DNA binding, 74 to 105%) together with two presumptive A. encheleia isolates. The fact that strain LMG 16330T of A. encheleia was the only type strain residing in DNA homology group II implies that HG11 and A. eucrenophila subgroup II should be classified in the species A. encheleia. Except for the somewhat aberrant phenotypic positions of HG11 strains LMG 13075 and LMG 13076, the establishment of DNA homology groups I and II was supported by the delineation of phena 1 and 2 (level of correlation, 90%), respectively, as revealed by numerical analysis of 136 phenotypic test results. These data indicate that A. eucrenophila and A. encheleia are phenotypically highly related but can be easily separated by testing the production of acid from D-cellobiose and lactose and the assimilation of D-cellobiose. Extended descriptions of both species are given. PMID- 9336924 TI - Aeromonas popoffii sp. nov., a mesophilic bacterium isolated from drinking water production plants and reservoirs. AB - We examined the taxonomic position of seven Aeromonas isolates, recovered from Flemish and Scottish drinking water production plants and reservoirs, which were previously recognized by numerical analysis of genomic AFLP fingerprints as members of an unknown Aeromonas taxon that most closely resembled the species Aeromonas bestiarum (DNA hybridization group [HG] 2). The new phenotypic and DNA DNA hybridization data obtained in this study show that the A. bestiarum-like strains constitute a separate Aeromonas species, for which the name Aeromonas popoffii sp. nov. is being proposed. The new species exhibited an internal DNA relatedness ranging from 79 to 100% and was 22 to 63% related to the type or reference strains of other Aeromonas spp. The highest DNA binding values were determined with A. bestiarum (51 to 63%), followed by Aeromonas hydrophila sensu stricto (HG1; 50 to 60%) and Aeromonas salmonicida (HG3; 39 to 55%). Although fingerprints generated by ribotyping and cellular fatty acid analysis often were highly similar, minor differences between the respective fingerprints were of significance for the differentiation of A. popoffii from its closest taxonomic neighbors, HG1, HG2, and HG3. Phenotypically, all seven strains of A. popoffii were positive for acid and gas production from D-glucose and glycerol, growth in KCN broth, arginine dihydrolase, DNase, Voges-Proskauer reaction, and resistance to vibriostatic agent O/129 and ampicillin but displayed negative reactions for production of urease, tryptophan deaminase, ornithine decarboxylase, and lysine decarboxylase (LDC). None of the strains displayed strong hemolytic activity. The lack of D-sucrose fermentation and LDC production and the ability to utilize DL lactate as the sole energy and carbon source were useful characteristics for the biochemical separation of A. popoffii from A. bestiarum. Other Aeromonas spp. could be differentiated phenotypically from the new species by at least two features. The chromosomal G+C content of A. popoffii ranges from 57.7 to 59.6 mol%. Strain LMG 17541 is proposed as the type strain. PMID- 9336925 TI - Reorganization of the genus Erythromicrobium: description of "Erythromicrobium sibiricum" as Sandaracinobacter sibiricus gen. nov., sp. nov., and of "Erythromicrobium ursincola" as Erythromonas ursincola gen. nov., sp. nov. AB - The results of investigations on the morphology, physiology, pigment composition, light-harvesting antenna and reaction center organization, and electron carriers of five Erythromicrobium representatives, and on phylogenetic relations among them, are summarized. On the basis of clear phenotypic differences and distinct phylogenetic positions shown by 16S ribosomal DNA analysis, the tentative species "Erythromicrobium sibiricum" and "Erythromicrobium ursincola" are formally described as the type species of two new genera: Sandaracinobacter sibiricus gen. nov., sp. nov., and Erythromonas ursincola gen. nov., sp. nov., respectively. The genus Erythromicrobium is at present composed of the type species, E. ramosum, and two species, "E. hydrolyticum" and "E. ezovicum," whose nomenclature is yet to be validated. All species studied group within the alpha-4 subclass of Proteobacteria. PMID- 9336926 TI - Discrimination of Acinetobacter genomic species by AFLP fingerprinting. AB - AFLP is a novel genomic fingerprinting method based on the selective PCR amplification of restriction fragments. The usability of this method for the differentiation of genomic species in the genus Acinetobacter was investigated. A total of 151 classified strains (representing 18 genomic species, including type, reference, and field strains) and 8 unclassified strains were analyzed. By using a single set of restriction enzymes (HindIII and TaqI) and one particular set of selective PCR primers, all strains could be allocated to the correct genomic species and all groups were properly separated, with minimal intraspecific similarity levels ranging from 29 to 74%. Strains belonging to genomic species 8 (Acinetobacter lwoffii sensu stricto) and 9 grouped together in one cluster. The closely related DNA groups 1 (Acinetobacter calcoaceticus), 2 (Acinetobacter baumannii), 3 and 13TU (sensu Tjernberg & Ursing 1989) were clearly distinguishable, with intraspecific linkage levels above 50%. Strains of the independently described genomic species 13BJ (sensu Bouvet & Jeanjean 1989) and 14TU linked together at a relatively low level (33%). Although a previous DNA-DNA hybridization study seemed to justify the unification of these genomic species, AFLP analysis actually divides the 13BJ-14TU group into three well-separated subgroups. Finally, four unclassified strains obtained from diverse sources and origins grouped convincingly together, with a similarity linkage level of approximately 50%. These strains showed no similarities in their AFLP patterns with any of the other 155 strains studied and may represent a thus-far undescribed Acinetobacter species. Based on these results, AFLP should be regarded as an important auxiliary method for the delineation of genomic species. Furthermore, because AFLP provides a detailed insight into the infraspecific structure of Acinetobacter taxa, the method also represents a highly effective means for the confirmation of strain identity in the epidemiology of acinetobacters. PMID- 9336927 TI - Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov. AB - We performed an integrated genotypic and phenotypic analysis of 128 strains of the genera Burkholderia, Ralstonia, and Pseudomonas in order to study the taxonomic structure of Burkholderia cepacia and its relationships with other Burkholderia species. Our data show that presumed B. cepacia strains isolated from cystic fibrosis patients belong to at least five distinct genomic species, one of which was identified as Burkholderia vietnamiensis. This group of five phenotypically similar species is referred to as the B. cepacia complex. The name Burkholderia multivorans is proposed for one of these genomic species, which was formerly referred to as B. cepacia genomovar II; the remaining B. cepacia groups are referred to as genomovars I, III, and IV, pending additional differential phenotypic tests. The role and pathogenic potential of each of these taxa, particularly in view of the potentially fatal infections in cystic fibrosis patients, need further evaluation. The data presented also demonstrate that Pseudomonas glathei and Pseudomonas pyrrocinia should be reclassified as Burkholderia species. PMID- 9336928 TI - Phenotypic and phylogenetic characterization of some Eubacterium-like isolates containing a novel type B wall murein from human feces: description of Holdemania filiformis gen. nov., sp. nov. AB - A group of Eubacterium-like strains (designated group S14), isolated from the feces of healthy people, was characterized by biochemical tests, fatty acid analysis, cell wall murein analysis, and 16S rDNA analysis. Our results indicate that group S14 is phylogenetically a member of the Clostridium subphylum of the gram-positive bacteria. Despite a phenotypic resemblance to the genus Eubacterium, group S14 was shown to be phylogenetically distantly related to the type species of the genus, Eubacterium limosum. Group S14 showed a specific phylogenetic association with Erysipelothrix rhusiopathiae. Group S14 resembled Erysipelothrix in possessing the uncommon type B cell wall murein. Structural analyses, however, revealed the presence of a previously unknown B1 delta (L-Ala) D-Glu-Gly-L-Lys murein type. Based on a 16S rRNA sequence divergence of greater than 10% with E. rhusiopathiae and the presence of a unique murein type, a new genus, Holdemania, is proposed for group S14, with one species, Holdemania filiformis. Type strain of H. filiformis is ATCC 51649. PMID- 9336929 TI - Mycobacterium novocastrense sp. nov., a rapidly growing photochromogenic mycobacterium. AB - A strain isolated from a biopsy sample taken from a slowly spreading skin granulation on a child's hand was found to have properties consistent with its classification in the genus Mycobacterium. An almost complete gene sequence of the 16S rRNA of the strain was determined following the cloning and sequencing of the amplified gene. The sequence was aligned with those available for mycobacteria, and phylogenetic trees were inferred with four tree-making algorithms. The organism, which formed a distinct phyletic line within the evolutionary radiation occupied by rapidly growing mycobacteria, was readily distinguished from members of validly described species of rapidly growing mycobacteria on the basis of its mycolic acid pattern and a number of other phenotypic features, notably its ability to form yellow pigmented colonies when incubated in the light. The name proposed for this new species is Mycobacterium novocastrense. The type strain is DSM 44203. PMID- 9336930 TI - Mycoplasma lagogenitalium sp. nov., from the preputial smegma of Afghan pikas (Ochotona rufescens rufescens). AB - Organisms with characteristics typical of mycoplasmas were isolated from the preputial smegma of Afghan picas (Ochotona rufescens rufescens). The results of growth inhibition tests, metabolic inhibition tests, and immunobinding assays showed that the isolated strains were identical and that they were distinct from previously described Mycoplasma, Entomoplasma, Mesoplasma, and Acholeplasma species. These organisms represent a new species, for which the name Mycoplasma lagogenitalium is proposed. M. lagogenitalium ferments glucose, does not hydrolyze arginine or urea, reduces tetrazolium chloride, possesses phosphatase activity, does not digest gelatin or casein, and does not produce films or spots. It lyses sheep erythrocytes and does not adsorb sheep, rabbit, or horse erythrocytes. Cholesterol or serum is required for growth. The growth temperature is 37 degrees C. The guanine-plus-cytosine content of the DNA is 23.0 +/- 1.0 mol%. The type strain is M. lagogenitalium 12MS (= ATCC 700289T). PMID- 9336931 TI - Sulfurospirillum arcachonense sp. nov., a new microaerophilic sulfur-reducing bacterium. AB - The isolation of a new motile, gram-negative, heterotrophic, sulfur-reducing, microaerophilic, vibrioid bacterium, strain F1F6, from oxidized marine surface sediment (Arcachon Bay, French Atlantic coast) is described. Hydrogen (with acetate as the carbon source), formate (with acetate as the carbon source), pyruvate, lactate, alpha-ketoglutarate, glutarate, glutamate, and yeast extract supported growth with elemental sulfur under anaerobic conditions. Apart from H2 and formate, the oxidation of the substrates was incomplete. Microaerophilic growth was supported with hydrogen (acetate as the carbon source), formate (acetate as the carbon source), acetate, propionate, pyruvate, lactate, alpha ketoglutarate, glutamate, yeast extract, fumarate, succinate, malate, citrate, and alanine. The isolate grew fermentatively with fumarate, succinate being the only organic product. Elemental sulfur and oxygen were the only electron acceptors used. Vitamins or amino acids were not required. The isolate was oxidase, catalase, and urease positive. Comparative 16S rDNA sequence analysis revealed a tight cluster consisting of the validly described species Sulfurospirillum deleyianum and the strains SES-3 and CCUG 13942 as the closest relatives of strain F1F6 (level of sequence similarity, 91.7 to 92.4%). Together with strain F1F6, these organisms form a novel lineage within the epsilon subclass of proteobacteria clearly separated from the described species of the genera Arcobacter, Campylobacter, Wolinella, and Helicobacter. Due to the phenotypic characteristics shared by strain F1F6 and S. deleyianum and considering their phylogenetic relationship, we propose the inclusion of strain F1F6 in the genus Sulfurospirillum, namely, as S. arcachonense sp. nov. Based on the results of this study, an emended description of the genus Sulfurospirillum is given. PMID- 9336932 TI - Terracoccus luteus gen. nov., sp. nov., an LL-diaminopimelic acid-containing coccoid actinomycete from soil. AB - A gram-positive, aerobic actinomycete was isolated from soil. Spherical cells of this organism occur singly or form packets, which may cluster. The diagnostic diamino acid of the cell wall peptidoglycan is LL-diaminopimelic acid. The predominate menaquinone is MK-8 (H4), and the main fatty acids are 13-methyl tetradecanoic acid and 12-methyl tetradecanoic acid. The diagnostic polar lipids are phosphatidylethanolamine and phosphatidylinositol. The DNA base composition is 73 mol% G + C. Comparison of 16S ribosomal DNA sequences showed that this isolate is a phylogenetic neighbor of Terrabacter tumescens and Intrasporangium calvum. Genotypic, chemotaxonomic, morphological, and physiological characteristics are used to describe a new genus and species, Terracoccus luteus gen. nov., sp. nov. The type strain is strain IMET 7848 (= DSM 44267). PMID- 9336933 TI - Meiothermus cerbereus sp. nov., a new slightly thermophilic species with high levels of 3-hydroxy fatty acids. AB - Strains of Meiothermus cerbereus sp. nov. were isolated from the hot springs within the Geysir geothermal area of Iceland. The strains of Meiothermus cerbereus produce red-orange-pigmented colonies, have an optimum growth temperature of about 55 degrees C, and have higher levels of 3-OH fatty acids than the strains of the other species of the genus Meiothermus. These strains, unlike all other strains of the species of the genus Meiothermus examined previously, required cysteine, thiosulfate, or thioglycolate for growth in liquid Thermus medium, but not in the corresponding medium solidified with agar. Several strains belonging to Meiothermus silvanus, isolated from Geysir, also required reduced sulfur compounds for growth in liquid medium, leading to the hypothesis that this requirement is not a taxonomic characteristic of the new species. The new species represented by strains GY-1T and GY-5 can be distinguished from the other species of the genus Meiothermus by biochemical characteristics, fatty acid composition, DNA-DNA reassociation values, and 16S ribosomal DNA sequence. The type strain for Meiothermus cerbereus is GY-1 (= DSM 11376). PMID- 9336934 TI - Polyphasic taxonomy of Nesterenkonia halobia. AB - A phenotypic study has been carried out on six moderately halophilic gram positive nonmotile cocci isolated from ponds of a saltern located in Huelva, Spain. These strains were examined for 150 morphological, physiological, biochemical, and nutritional traits and showed phenotypic characteristics similar to those of Nesterenkonia halobia (formerly Micrococcus halobius). The guanine plus-cytosine (G + C) content of their DNA ranged between 70 and 72 mol%, values quite similar to those described for N. halobia (71.5 mol%). The 16S rDNA sequence analysis of one representative isolate showed that it is phylogenetically quite close to N. halobia, within the high-G + C-content gram positive branch. DNA-DNA hybridization experiments showed a high degree of homology (72 to 100%) among the six isolates and the type strain N. halobia ATCC 21727. All data demonstrate quite clearly that the six isolates are members of the species N. halobia. Since this species was described on the basis of a single strain isolated from unrefined solar salt, and its description is not complete (especially in the utilization of different compounds), our study contributes to a better description of the moderate halophile N. halobia. PMID- 9336935 TI - A novel pathogenic taxon of the Mycobacterium tuberculosis complex, Canetti: characterization of an exceptional isolate from Africa. AB - In an attempt to characterize an unusual mycobacterial strain isolated from a 2 year-old Somali patient with lymphadenitis, we applied various molecular methods not previously used for the taxonomic classification of mycobacteria. This isolate, designated So93, did not differ from Mycobacterium tuberculosis in the biochemical tests and in its 16S rRNA sequence, but produced smooth and glossy colonies, which is highly exceptional for this species. This smooth phenotype was unstable and switched nonreversibly to a rough colony morphology with a low frequency. The two colony types were equally virulent for the guinea pig, exhibiting characteristic tuberculous disease. Both morphotypes had shorter generation times than the M. tuberculosis reference laboratory strain H37Rv and clinical isolates of M. tuberculosis and Mycobacterium bovis. Furthermore, the So93 isolate differed from all M. tuberculosis complex strains described thus far by having only a single copy of insertion sequence IS1081, an unusual composition of the direct repeat cluster, and a characteristic phenolic glycolipid and lipooligosaccharide. This glycolipid had previously been observed only in a smooth isolate of M. tuberculosis obtained in 1969 by Canetti in France. Analysis of the Canetti strain showed that it shared virtually all genetic properties characteristic of So93, distinguishing these two strains from the known M. tuberculosis complex taxa, M. tuberculosis, Mycobacterium africanum, M. bovis, and Mycobacterium microti. The natural reservoir, host range, and mode of transmission of the group of bacteria described in this paper are presently unknown. This study, partly based on not previously used molecular criteria, supports the idea that the established members within the M. tuberculosis complex and the newly described Canetti grouping should be regarded as a single species, which likely will be designated "M. tuberculosis". PMID- 9336936 TI - Rapid identification of heterotrophic, thermophilic, spore-forming bacteria isolated from hot composts. AB - The restriction enzyme profiles of 16S ribosomal DNAs (rDNAs) amplified by PCR from thermophilic heterotrophic bacterial strains isolated from composts were compared with those of reference strains. This allowed us to assign all but 1 of 16 strains to four different Bacillus species (namely, Bacillus stearothermophilus, Bacillus pallidus, Bacillus thermoglucosidasius, and "Bacillus thermodenitrificans"). This study showed that PCR restriction analysis of 16S rDNA contributes to rapid and reliable identification of newly isolated strains belonging to recognized species. PMID- 9336937 TI - Proposal to reclassify Zoogloea ramigera IAM 12670 (P. R. Dugan 115) as Duganella zoogloeoides gen. nov., sp. nov. AB - The taxonomic position of a misclassified strain, Zoogloea remigera IAM 12670T (= ATCC 25925T = P. R. Dugan 115T), was reevaluated. A phylogenetic analysis based on 16S ribosomal rDNA sequences revealed that this organism was located in the beta subclass of the class Proteobacteria with members of the genus Telluria as its closest relatives. On the basis of phenotypic and phylogenetic information, we propose that this organism should be reclassified in a new taxon with the name Duganella zoogloeoides gen. nov., sp. nov. PMID- 9336939 TI - Reassessment of the taxonomic position of Rickettsiella grylli. AB - We determined the 16S rRNA gene sequence of Rickettsiella grylli, an intracellular parasite of Gryllus bimaculatus and related species of crickets. Phylogenetic inferences made from alignment of this sequence with the sequences of other bacteria demonstrated that R. grylli is most closely related to Coxiella burnetii and Legionella species in the gamma subclass of the phylum Proteobacteria. R. grylli was previously thought to be related to members of the order Rickettsiales, but the representatives of this order have been shown to be members of the alpha 1 subclass of the Proteobacteria. Our results indicate that R. grylli should be removed from the order Rickettsiales. PMID- 9336938 TI - Phylogenetic relationships of Salmonella typhi and Salmonella typhimurium based on 16S rRNA sequence analysis. AB - The 16S rRNA gene sequences of Salmonella typhi and Salmonella typhimurium were amplified by PCR, cloned, and sequenced. These sequences were analyzed by comparison with reference organisms from the family Enterobacteriaceae. Both S. typhi and S. typhimurium belong to the gamma subdivision of the class Proteobacteria. PMID- 9336940 TI - Isolation of an aceticlastic strain of Methanosarcina siciliae from marine canyon sediments and emendation of the species description for Methanosarcina siciliae. AB - A newly described strain of the genus Methanosarcina was isolated from submarine canyon sediments and is shown by comparative sequence analyses of 16S ribosomal DNA and the gene encoding methyl coenzyme M reductase, mcrI, to be a strain of Methanosarcina siciliae. Morphological and physiological characteristics are described. In contrast to the two previously described strains that grow exclusively on methanol, methylamines, and dimethylsulfide, M. siciliae C2J is also capable of growth on and methanogenesis from acetate. We propose that the species description for M. siciliae be amended to include aceticlastic strains. PMID- 9336941 TI - PCR-restriction fragment length polymorphism analysis of genes coding for 16S rRNA in Veillonella spp. AB - Restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal DNA (16S rDNA PCR-RFLP) was used to generate restriction profiles of the American Type Culture Collection type strains of the genus Veillonella, i.e., V. atypica, V. caviae, V. criceti, V. dispar, V. parvula, V. ratti, and V. rodentium. Whole cell protein profiles were obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for comparative purposes. The 16S rRNA genes were amplified by PCR, and RFLP analysis of the 16S rDNA was performed with MnlI and Sau3AI. MnlI produced six RFLP patterns for seven type strains, since the patterns for V. atypica and V. caviae were the same. RFLP patterns with Sau3AI could distinguish between V. atypica and V. caviae. The type strains of Veillonella species were easily distinguished by 16S rDNA PCR-RFLP. PMID- 9336942 TI - Validation of the publication of new names and new combinations previously effectively published outside the IJSB. List No. 63. PMID- 9336944 TI - Bacterial genetic loci implicated in the Pseudomonas putida GR12-2R3--canola mutualism: identification of an exudate-inducible sugar transporter. AB - Pseudomonas putida GR12-2R3 promotes the emergence and growth of diverse plant species. Analyses of TnphoA insertion mutations are revealing bacterial characteristics pertinent to the plant-microbe interaction. Pseudomonas putida PG269 is a TnphoA insertion derivative of GR12-2R3 that expresses canola seed exudate-inducible alkaline phosphatase (PhoA) activity. It promoted the growth of canola roots, as well as strain GR12-2R3, and outgrew its parent when they were cocultured in the presence of canola roots or in liquid seed exudate medium. (In contrast, mutant PG126 failed to promote canola root growth and was outgrown by its parent strain.) The PhoA activity of strain PG269 was induced by glucosamine and other sugars; glucosamine inhibited the growth of strain GR12-2R3 and stimulated the growth of strain PG269. Strain PG269 contained two TnphoA insertions: seiA1::TnphoA and seiB1::TnphoA. Strain PG312, which contained only insertion seiA1::TnphoA, shared all aspects of the PG269 phenotype, except the ability to outcompete strain GR12-2R3 during coculture. Insertion seiA1::TnphoA interrupted an open reading frame related in sequence to members of the MalF family of sugar transporter subunits. The PhoA-inducing fraction of canola seed exudate was hydrophilic, low in molecular weight, and heat stable. It cochromatographed with basic amino acids and amino sugars, and was inactivated by strains GR12-2R3 and PG269. Gene seiA may encode a subunit of an ABC transporter with broad specificity for glucose and related sugars whose expression can be induced by exudate sugars. PMID- 9336945 TI - Mapping of sequences required for the translation of the beta subunit of Escherichia coli RNA polymerase. AB - Previous experiments using expression plasmids which overproduce the beta and beta' subunits of Escherichia coli RNA polymerase suggested that regions considerably upstream of the start of the rpoB gene, which encodes the beta subunit, are required for its efficient synthesis. To further delineate the required regions, a collection of genetic constructs that contained varying amounts of the region either upstream or downstream of the translational start of rpoB was assembled. Measurements of beta and beta' synthesis and rpoB mRNA production from a series of rpoBC expression plasmids indicated that sequences extending more than 43 bp but less than 79 bp upstream of rpoB are required for the efficient translation of rpoB mRNA. This result was confirmed by beta galactosidase measurements from a series of rpoB-lacZ fusions that have the same set of end points upstream of rpoB as the expression plasmids. A second set of gene fusions containing differing amounts of the sequence distal to the start of rpoB fused in frame to lacZ revealed that more than 29 bp but less than 70 bp of rpoB was required for efficient translation. PMID- 9336947 TI - Biodegradation of groundwater pollutants by a combined culture of Mycobacterium vaccae and a Rhodococcus sp. AB - The catabolism of selected groundwater pollutants by a combined culture of Mycobacterium vaccae and a Rhodococcus sp. (strain R-22) was investigated. The M. vaccae-R-22 combined culture was five times more effective in mineralizing benzene than either organism alone. Mycobacterium vaccae oxidized benzene to phenol, and R-22 catabolized the phenol to cellular components and CO2. Benzene did not support growth of M. vaccae, R-22, or the combined culture. Optimization of ratios of the two species indicated that the maximum mineralization of benzene occurred at an initial ratio of 75% M. vaccae to 25% R-22. Cell fractionation of the combined culture after mineralization of [U-14C]benzene indicated that 10% of the benzene carbon was incorporated into cell material, and of this 45% was present in protein and 20% in nucleic acids. This suggested that minimally one species could utilize the products of benzene as a nutrient source. The M. vaccae R-22 combined culture catabolized ethylbenzene and chlorobenzene without the accumulation of phenolic intermediates, which are inhibitory to M. vaccae's ability to degrade the parent compounds. This study demonstrates that defined mixed cultures may be useful in studying the effects of environmental pollutant degradation on microbial ecosystems and mineralization of these pollutants by the ecosystem. PMID- 9336948 TI - Rapid identification of Medicago nodulating strains by using two oligonucleotide probes complementary to 16S rDNA sequences. AB - Symbiotic bacteria associated with the Medicago genus are separated into two closely related species named Sinorhizobium meliloti and Sinorhizobium medicae. To discriminate rapidly between these two bacterial species, two 15-base DNA probes, 16Smfs and 16Smed, were designed from the alignment of 16S rDNA sequences to differentiate S. meliloti from S. medicae. Their specificities were evaluated by dot-blot hybridization experiments on 25 reference strains representing 13 species of Rhizobium and Sinorhizobium, and by comparison with all 16S rDNA sequences available in the GenBank data base. No cross-reaction was found with 16Smed, which was thus considered species specific for S. medicae. By contrast, as expected according to the 16S rDNA sequence alignment, the labeled 16Smfs probe cross-hybridized with the DNAs of S. meliloti, Sinorhizobium fredii, and Sinorhizobium saheli but not with the DNA of S. medicae. Since S. saheli and S. fredii do not nodulate Medicago, 16Smed and 16Smfs can be routinely used to characterize the two Sinorhizobium species nodulating Medicago from pure cultures or from Medicago root nodules. Fifty strains isolated from eight annual Medicago species were then characterized by using colony hybridizations. Sinorhizobium meliloti was more frequently obtained (> 80% isolates) than was S. medicae. Both Sinorhizobium species seemed to be trapped by annual Medicago and no plant-host specificity was detected. PMID- 9336949 TI - On the relation of colony variants to the time dependency of colony formation during adaptive mutation of Escherichia coli FC40. AB - When Escherichia coli FC40 formed adaptive Lac+ revertants on a selective agar medium containing lactose as the carbon source, the colonies which accumulated over several days were of two readily distinguishable types. Colonies of both types appeared both early and late on the plates. Cells of colonies that appeared early and late on the plates, irrespective of the type, when grown in liquid medium and replated, all formed colonies on the selective medium within 48 h. Cells of each colony type gave rise to colonies of both types and attempts to isolate cells of each type in pure culture were unsuccessful. It was concluded that the presence of two colony types in the cultures plated did not contribute to the observed time dependency of colony formation during adaptive mutation. The proportions of the two colony types arising from cultures of the Lac+ revertants varied from culture to culture. PMID- 9336954 TI - Development of a gas chromatographic-mass spectrometric drug screening method for the N-dealkylated metabolites of fentanyl, sufentanil, and alfentanil. AB - A sensitive, specific urinary assay for fentanyl, sufentanil, and alfentanil based on their N-dealkylated metabolites is described. Norfentanyl, norsufentanil noralfentanil, and 2H5-norfentanyl are synthesized and characterized by standard analytical techniques. Derivatization of these secondary amines to yield the pentafluorobenzamides produces stable products with good gas chromatographic properties and unique, high-mass fragments in their mass spectra. These properties are utilized to develop a drug screening procedure based on gas chromatography-mass spectrometry to detect these major metabolites in human urine. The metabolites are isolated from urine samples by a liquid-liquid extraction procedure. The method allows for detection of metabolite concentrations as low as 0.3 ng/mL. PMID- 9336955 TI - Stress, neuromotor noise, and human performance: a theoretical perspective. AB - A new theory on stress and human performance is proposed in which physical and cognitive stressors enhance the level of neuromotor noise in the information processing system. The neuromotor noise propagates in time and space. A 2nd assumption states that such noise facilitates easy tasks but disrupts complex tasks. In 4 experiments, 2 graphic tasks (number writing and graphic aiming) were crossed with 2 stressors (cognitive stress from a dual-task situation and physical stress in the form of loud auditory noise). Reaction time (RT), movement time (MT), and axial pen pressure were measured. In the RT phase, stress was predicted to lead to decreased RT with easy tasks and to increased RT with difficult tasks. In the execution phase, biomechanical adaptation to enhanced levels of noise was expected to manifest in higher levels of limb stiffness. In all 4 experiments, an increase of axial pen pressure with higher levels of stress evidenced the generality of biomechanical adaptation as a response to stress. RT and MT showed differential effects among the 4 experiments. PMID- 9336956 TI - A belongingness principle of motion perception. AB - Four experiments are reported that investigated the role of the perceived coplanarity of a moving target with respect to a frame of reference in the third dimension on the perceived path of that target. When a target dot and small moving frame appeared coplanar, the dot's perceived trajectory was governed entirely by its changing position relative to the moving frame. However, when the target and a large stationary frame appeared in a different plane than the small moving frame, the motion of the dot was seen independently of the moving frame. The results support a belongingness principle of motion perception: The displacement of an object relative to a frame of reference to which it belongs governs its perceived path of motion. PMID- 9336957 TI - Postural dynamics and the preferred critical boundary for visually guided reaching. AB - How do people choose an action to satisfy a goal from among the actions that are afforded by the environment? In 3 experiments the action modes used by actors to reach for a block placed at various distances from them were observed. In each experiment, when actors were not restricted in how they could reach for the object, the transition from their reaching using only arm extension to a mode of reaching in which they used the upper torso to lean forward occurred at closer distances than each actor's absolute critical boundary, beyond which the former action was no longer afforded. In Experiments 2 and 3 actors' seated posture was varied so that the effect of postural dynamics on the distance at which actors actually chose to make the transition between action modes, the preferred critical boundary, could be examined. The results are consistent with the proposal that the preferred critical boundary reflects the relative comfort of available modes of reaching. PMID- 9336958 TI - Shifting attention in visual space: tests of moving-spotlight models versus an activity-distribution model. AB - Participants were induced to concentrate preparatory attention at a central location, to identify a letter there, to identify a 2nd letter to the extreme left or right of a central horizontal range of 5 locations, and then to identify a 3rd letter at 1 of the central 5 locations. Analog and discrete versions of the moving-spotlight model predict that response times to the 3rd letter will be most rapid at the location of the 2nd letter, whereas an activity-distribution model predicts that the most rapid responses to the 3rd letter will be at the central location, where preparatory attention is strongest. The data from 3 experiments, taken together, are inconsistent with the moving-spotlight models and are consistent with the activity-distribution model. PMID- 9336959 TI - Sources of the attentional blink during rapid serial visual presentation: perceptual interference and retrieval competition. AB - Observers watched for 1 or 2 colored words as targets presented in lists of distractor strings (10 items/s). Identification of 1 target (T1) temporarily reduced the accuracy of reporting a 2nd target (T2). This attentional blink (AB) effect was most pronounced when T1 and T2 occurred close together in time. Use of recognition tests (instead of recall) improved performance but did not eliminate the AB effect. The AB effect was found with both word and nonword distractors, a smaller AB effect was found with consonant string distractors, and the AB effect was substantially attenuated with strings of unfamiliar characters (a false font). Analyses of errors indicated that the 2nd target is frequently replaced or corrupted by the following distractor during the blink. The AB effect appears to result from both attentional and mnemonic processes. PMID- 9336960 TI - Effect of frequency ratio on infants' and adults' discrimination of simultaneous intervals. AB - Effects of frequency ratio simplicity on infants' and adults' processing of simultaneous pitch intervals with component sine wave tones were tested. Both infants and adults showed superior performance at detecting a change from a perfect 5th (2:3) to either a tritone (32:45) or a minor 6th (5:8) interval than at detecting the reverse discriminations (minor 6th or tritone to perfect 5th). Similarly, both infants and adults showed superior performance at detecting a change from an octave (1:2) to either a major 7th (8:15) or a minor 9th (15:32) interval than at detecting the reverse discriminations. In combination with previous findings of infants' superior discrimination of tone sequences with prominent perfect 5th intervals, these results suggest that both simultaneous and sequential intervals with simple ratios are easy to process early in development. PMID- 9336961 TI - Coordination dynamics of learning and transfer: collective and component levels. AB - The dynamics of learning a new coordinated behavior was examined by requiring participants to perform a visually specified phase relationship between the hands. Results showed that learning may involve qualitative or quantitative alterations in the layout of the coordination dynamics depending on whether such dynamics are bistable or multistable before exposure to the learning task. In both cases, the process stabilized the to-be-learned behavior and its symmetry partner, even though the latter had not actually been practiced. Kinematic analyses of hand motion showed that previously existing coordination tendencies were exploited during learning in order to match visual requirements. These findings and the concepts presented here provide a framework for understanding how learning occurs in the context of previous experience and allow individual differences in learning to be tackled explicitly. PMID- 9336962 TI - Perceptual localization of surface position. AB - In 2 experiments, observers were required to identify corresponding points on an object viewed from multiple orientations. On each trial, a surface was presented initially with a single target location marked by a small dot. Following a brief blank interval, the same surface was presented again at a different orientation. The observer was required to position an adjustable probe dot in this 2nd display to match the location of the target in the 1st view. Under optimal conditions, the variance in their settings over multiple trials was just a few minutes of arc, though these errors varied significantly with the structural complexity of the depicted surface. PMID- 9336963 TI - Object-based facilitation and inhibition from visual orienting in the human split brain. AB - Object-based attention was examined in 2 split-brain patients. A precued object could move within a visual field or cross the midline to the opposite field. Normal individuals show an inhibition in detecting signals in the cued object whether it moves within or between fields. Both patients showed this effect when the cued object moved within a visual field. When it crossed the midline into the opposite visual field, however, detection was faster in the cued box. These results reveal both facilitatory and inhibitory effects on attention that are object based and may last for several hundred milliseconds. However, the inhibition requires an intact corpus callosum for interhemispheric transfer, whereas the facilitation is transferred subcortically. PMID- 9336964 TI - Process dissociation, cognitive architecture, and response time: comments on Lindsay and Jacoby (1994) AB - Process dissociation is based on 2 assumptions about the processes being dissociated: invariance of the processes across situations, and stochastic independence of the processes. In a recent application of process dissociation to the Stroop task (D. S. Lindsay & L. L. Jacoby, 1994), both of those assumptions were violated. It is argued that these violations were due to (a) an oversimplification of the processing architecture that ignores common stages such as guessing and response selection, (b) an assumption that the more automatic process (word reading) dominates over the intended process (color naming) in determining responses, and (c) an assumption that switching from the more common speeded response instruction (measuring speed) to a deadline response instruction (measuring accuracy) does not change processing. General implications for applying process dissociation to dynamic tasks are discussed. PMID- 9336966 TI - Perceptual and mental mixtures in odor and in taste: are there similarities and differences between experiments or between modalities? Reply to Schifferstein (1997) AB - D. Algom and W. S. Cain (1991b) found relative invariance in the pattern of judgments of perceptual and mental mixtures of banana and grass odors. Invariance held both for judgments of total intensity and for those of an individual constituent. For 2 tastes, H. N. J. Schifferstein (1997) found a pattern with both similarities to and differences from D. Algom and W. S. Cain's. A key difference lay in finding more symmetry of masking in mental mixtures than in perceptual mixtures. H. N. J. Schifferstein concluded from this alone that any similarity between the perceptual and mental arose from knowledge of "mixture suppression." The authors of this article do not refute the possibility; however, they reject the premise that a statistically reliable difference between the perceptual and the imaginal rules out imagery. The authors review relevant considerations and find no a priori reason to assume that what held for attributes in taste will hold for odors. An approach to resolve the issue is also suggested. PMID- 9336968 TI - Purification of a factor that enhances the antibacterial activity of beta-lactams against methicillin-resistant Staphylococcus aureus: its identification as undecaphosphotungstate. AB - Previously, a factor (Factor T) was found in aged mixtures of tungstate and phosphate, which greatly enhances the antibacterial effects of beta-lactam antibiotics on methicillin-resistant Staphylococcus aureus. Factor T was purified and identified as undecaphosphotungstate ([PW11O39]7-). Undecasilicotungstate ([SiW11O39]8-), a compound closely related to undecaphosphotungstate, showed a similar enhancing effect. PMID- 9336967 TI - Synthesis and DNA binding studies of cobalt (III) mixed-polypyridyl complex. AB - The complex of Co(phen)2dppz3+ (where phen is ophenanthroline, and dppz is dipyrido [3, 2-a: 2', 3'-c] phenazine) has been synthesized. This complex was characterized by elemental analysis, molar conductivity, IR and 1H NMR spectroscopy. The interaction of the complex with calf thymus DNA has been studied using absorption and emission spectroscopy, DNA melting techniques, cyclic voltammetric, viscosity, and electrophoresis measurements. The compound shows absorption hypochromicity, fluorescence enhancement, DNA melting temperature, and the specific viscosity increased when binding to calf thymus DNA. CV measurement shows that the shifts in oxidation-reduction potential and change in peak current with addition of DNA. The complex is also shown to be more efficient photosensitisers for single strand breaks in plasmid DNA. PMID- 9336969 TI - Glucose-lowering properties of vanadium compounds: comparison of coordination complexes with maltol or kojic acid as ligands. AB - Bis(kojato)oxovanadium(IV) [abbreviated VO(ka)2], a close chemical analog of the insulin-mimetic lead compound bis(maltolato)oxovanadium(IV)--abbreviated BMOV or VO(ma)2--is reported and its reaction chemistry and insulin-mimetic properties are presented. VO(ka)2 [log K1 = 7.61(10), log K2 = 6.89(6), log beta 2 = 14.50(16)] has a reaction chemistry which directly parallels that of VO(ma)2. In aqueous solution it is more slowly oxidized by molecular oxygen to [VO2(ka)2]- than is VO(ma)2 to [VO2(ma)2]-. Variable pH electrochemistry and variable pH 51V NMR of solutions of VO(ka)2 are presented and contrasted with the corresponding results for VO(ma)2. Time course studies (24 hr) in STZ-diabetic rats following the oral or i.p. administration of VO(ka)2, VO(ma)2, VO2+ (vanadyl) as vanadyl sulfate (VOSO4), and [VO2(ma)2]- as its [NH4]+ salt have been performed, as have chronic oral studies comparing VO(ka)2 and VO(ma)2 over a six week period. In all studies, the most potent form of vanadium was the neutrally charged, water soluble, complex VO(ma)2. PMID- 9336970 TI - Experimental antitumor activity of the Ce(IV)-mitoxantrone complex and its interaction with deoxyribonucleic acid. AB - The Ce(IV)-mitoxantrone complex exhibits a higher lethality to Ehrlich ascites tumor cells than that of the free drug and shows stronger inhibition ability on the DNA synthesis of the tumor cells. Thus the Ce(IV)-mitoxantrone complex may become a more potent antitumor drug than mitoxantrone. The different interaction model of mitoxantrone and its Ce(IV) complex with DNA were studied by the methods of spectroscopy, electrochemistry, and electrophoresis. Ce(IV) ions chelate with oxygens of the hydroxyl groups at the 1,4 position and the carbonyl function on C 9 and C-10, then intercalate into the base pairs of DNA together. The complexation of Ce(IV) gives rise to more compact binding of mitoxantrone with DNA, and leads to an additional change on the normal conformation and the double helical structure of DNA; this may be related to the more stronger action of the complex on DNA synthesis and survival of cultured tumor cells. PMID- 9336971 TI - Effect of calcium ions on rat osseous plate alkaline phosphatase activity. AB - Rat osseous plate alkaline phosphatase is a metalloenzyme with two binding sites for Zn2+ (sites I and III) and one for Mg2+ (site II). This enzyme is stimulated synergistically by Zn2+ and Mg2+ (Ciancaglini et al., 1992) and also by Mn2+ (Leone et al., 1995) and Co2+ (Ciancaglini et al., 1995). This study was aimed to investigate the modulation of enzyme activity by Ca2+. In the absence of Zn2+ and Mg2+, Ca2+ had no effects on the activity of Chelex-treated, Polidocanol solubilized enzyme. However, in the presence of 10 microM MgCl2, increasing concentration of Ca2+ were inhibitory, suggesting the displacement of Mg2+ from the magnesium-reconstituted enzyme. For calcium-reconstituted enzyme, Zn2+ concentrations up to 0.1 microM were stimulatory, increasing specific activity from 130 U/mg to about 240 U/mg with a K0.5 = 8.5 nM. Above 0.1 microM Zn2+ exerted a strong inhibitory effect and concentrations of Ca2+ up to 1 mM were not enough to counteract this inhibition, indicating that Ca2+ was easily displaced by Zn2+. At fixed concentrations of Ca2+, increasing concentrations of Mg2+ increased the enzyme specific activity from 472 U/mg to about 547 U/mg, but K0.5 values were significantly affected (from 4.4 microM to 38.0 microM). The synergistic effects observed for the activity of Ca2+ plus magnesium reconstituted enzyme, suggested that these two ions bind to the different sites. A model to explain the effect of Ca2+ on the activity of the enzyme is presented. PMID- 9336972 TI - Bridged cobalt amine complexes induce DNA conformational changes effectively. AB - Cobalt(III) hexaamine ion [Co(NH3)6]3+ is known to facilitate the transition of B to Z-DNA or B- to A-DNA depending on the DNA sequences. Specific interactions are found between the amines of [Co(NH3)6]3+ and DNA atoms of A-DNA or Z-DNA. Bridged Co(III)pentaamine complexes, with multiple amine groups arranged in a rigid framework, may enhance the effectiveness of the conformational transition by occupying simultaneously two of the Co(III) hexaamine binding sites. Therefore, the imidazole-bridged Co(III)pentaamine complexes have been synthesized and their interactions with DNA oligonucleotides investigated by circular dichroism (CD) and NMR spectroscopy. CD studies of the titrations of d(A2C15G15T2) with [(NH3)5Co(Im)Co(NH3)5]Br5 and [(NH3)5Co(Im)2Co(NH3)4]Br7 showed that the former metal compound indeed is more effective than [Co(NH3)6]3+ in inducing the transition from B- to A-DNA. The conversion of B- to A-DNA was also supported by one- and two-dimensional NMR studies. Similarly for the titrations of poly(dC-dG). poly(dC-dG) and d(m5C-G)15 with these two bridged Co(III) complexes, efficient induction of Z-DNA was observed. Our studies suggest that bridged Co(III)pentaamine complexes may be useful agents for studying nucleic acid structures. PMID- 9336973 TI - Thermodynamic factors controlling the interaction of quinoline antimalarial drugs with ferriprotoporphyrin IX. AB - The interaction of a variety of quinoline antimalarial drugs as well as other quinoline derivatives with strictly monomeric ferriprotoporphyrin IX [Fe(III)PPIX] has been investigated in 40% aqueous DMSO solution. At an apparent pH of 7.5 and 25 degrees C, log K values for bonding are 5.52 +/- 0.03 (chloroquine), 5.39 +/- 0.04 (amodiaquine), 4.10 +/- 0.02 (quinine), 4.04 +/- 0.03 (9-epiquinine), and 3.90 +/- 0.08 (mefloquine). Primaquine, 8 hydroxyquinoline, 5-aminoquinoline, 6-aminoquinoline, 8-aminoquinoline, and quinoline exhibit no evidence of interaction with Fe(III)PPIX. The enthalpy and entropy changes for the interaction of quinolines with Fe(III)PPIX, as determined from the temperature dependence of the log K values, exhibit a compensation phenomenon that is suggestive of hydrophobic interaction. This is supported by the finding that the interactions of chloroquine and quinine with Fe(III)PPIX are weakened by increasing concentrations of acetonitrile. Interactions of chloroquine, quinine, and 9-epiquinine with Fe(III)PPIX are shown to remain strong at pH 5.6, the approximate pH of the food vacuole of the malaria parasite which is believed to be the locus of drug activity. Implications for the design of antimalarial drugs are briefly discussed. PMID- 9336974 TI - Palladium(II) complexes of 2-acetylpyridine N(4)-methyl, N(4)-ethyl and N(4) phenyl-thiosemicarbazones. Crystal structure of chloro(2-acetylpyridine N(4) methylthiosemicarbazonato) palladium(II). Synthesis, spectral studies, in vitro and in vivo antitumour activity. AB - The reactions of 2-acetylpyridine N(4)-methyl, (HAc4Me) N(4)-ethyl (HAc4Et) and N(4)-Phenyl (HAc4Ph) thiosemicarbazone with palladium(II) were studied. The ligands and the palladium(II) complexes have been characterized by spectroscopic techniques. The structure of [Pd(Ac4Me)Cl] has been determined by single-crystal x-ray diffraction. The protonation constants of HAc4Me and HAc4Et, Ka1 and Ka2, were determined by spectrophotometry. The effect of palladium compounds on DNA synthesis of P388 and L1210 cell cultures is also reported. Some of these compounds increased the life span of mice bearing tumors. PMID- 9336975 TI - Acetate-specific stress response in acetate-resistant bacteria: an analysis of protein patterns. AB - Many metabolic byproducts have toxic effects on bacteria, and acetic acid is an excellent model for such molecules. The negative effects of acetate, which include decreased growth rates and specific productivities, appear for Escherichia coli at acetate concentrations lower than 5 g/L. Acetic acid bacteria, however, are naturally resistant to the detrimental effects of acetate in their surroundings; they remain active at acetate levels well over 40 g/L. This study investigated the response to acetate challenges by the naturally acetate-resistant bacteria Acetobacter aceti and Gluconobacter suboxydans to learn more about possible mechanisms of tolerance to otherwise toxic low molecular weight metabolites. Growth studies showed that the resistant bacteria grow more slowly in the presence of acetate but are not slowed nearly so much as is E. coli. In addition, two-dimensional gel electrophoresis (2DE) was applied to study the relative protein patterns of acetate-resistant bacteria during growth in the presence and absence of acetate. In each organism, growth in acetate containing medium led to elevated levels of many stress response proteins. 2DE analysis of heat-shocked cultures was used to determine which were nonspecific. Elimination of those proteins that were also amplified following heat shock left only eight proteins, here designated acetate-specific stress proteins (Asps), which are overexpressed specifically in response to acetate. Three of these, AspA, AspB, and AspC, appear to be analogous in the two bacterial strains studied, based on their apparent pIs and molecular weights. PMID- 9336976 TI - Porous alginate--poly(ethylene glycol) entrapment system for the cultivation of mammalian cells. AB - A novel gel entrapment method has been developed where macropores are created within alginate beads to provide an environment for high-density growth of mammalian cells. The method takes advantage of an interaction between poly(ethylene glycol) (PEG) and alginate to provide a network of pores within the bead for growth while the surrounding alginate matrix retains the integrity of the bead and minimizes cell leakage. Hybridomas were grown to a density approaching 10(8) cells/mL of beads in this system, while conventional alginate restricted growth to a maximum of 2 x 10(7) cells/mL of beads. In addition, cell leakage was minimal even at high cell densities, which was not the case with the conventional alginate system. Study of the conventional system determined that cell growth was limited by the alginate matrix; increasing the alginate concentrations resulted in lower final cell densities. In contrast, the PEG alginate system permits growth in pores so the alginate matrix serves only as a structural matrix for cells. The pore size can be varied as a function of PEG concentration (10-20 wt % PEG) to provide radially defined areas for cell growth and radial diffusion pathways for nutrients/products in the adjacent alginate matrix. Because the PEG-alginate entrapment process does not require additional chemical reactions or temperature changes, the system offers a simple alternative to attain high cell densities in an immobilized bead system. As an illustration of the concept, cells entrapped in this system were grown to high density in both batch and perfusion modes for the production of monoclonal antibodies. Using the suspension batch culture as the base case, the specific monoclonal antibody production rate increased 1.6-fold for the slower growing batch-immobilized culture and 3-fold for the immobilized perfusion culture. PMID- 9336977 TI - Immobilization of beta-fructofuranosidases from Aspergillus on methacrylamide based polymeric beads for production of fructooligosaccharides. AB - beta-Fructofuranosidases from Aspergillus niger ATCC 20611 and Aspergillus japonicus TIT-KJ1 were purified and immobilized covalently onto methacrylamide based polymeric beads. The porous, oxriane-containing support was reactive and could bind enzymes in a buffered solution at room temperature with a density up to 0.4 mg of protein g-1 of support with 100% immobilized yield. Neither the optimum temperature for the highest enzymatic activities nor the batch reaction pattern for fructooligosaccharides formation catalyzed by beta fructofuranosidases was changed by immobilization. The amount of fructooligosaccharides produced from 50% (w/w) sucrose solution using the prepared immobilized enzymes was determined to be approximately 60% of the total sugars in the reaction mixtures. This level of fructooligosaccharides produced by the immobilized enzymes was comparable to that resulting from processes employing other immobilized biocatalysts as shown in the literature. The fraction of total fructooligosaccharides presented in the final mixture increased with the initial sucrose concentration, while fractions of glucose and fructose decreased with an increase sucrose concentration. PMID- 9336978 TI - Manipulation of lyophilization-induced phase separation: implications for pharmaceutical proteins. AB - Lyophilization, or freeze-drying, of pharmaceutical proteins is often the only processing method that provides requisite long-term product stability. Freezing and drying, however, can cause acute damage to proteins. To alleviate damage, formulations frequently include protein stabilizers (often polymers and/or sugars), as well as buffering salts and "inert" bulking agents. While great efforts are placed on developing a formulation and suitable lyophilization cycle, incompatibilities among components through freezing and drying have been almost completely ignored. We demonstrate that solutions of poly(ethylene glycol) (PEG) and dextran, initially below critical concentrations for phase separation, do indeed experience a liquid-liquid phase separation induced by freeze concentration during the lyophilization cycle. The separation is shown to evolve with annealing at -7 degrees C and can be effectively inhibited simply by replacing NaCl with KCl in the formulation buffer. In addition, we show that phase separation causes unfolding of a model protein, recombinant hemoglobin, when freeze-dried in the PEG/dextran system. When the phase separation is averted by switching to KCl, the protein structural damage is also avoided. Measurements of pH in the frozen solutions show that the structural damage is not a result of pH changes. We suggest that KCl forms a glass with rapid cooling which kinetically prevents the phase separation and thus the protein structural damage. PMID- 9336979 TI - Fast ion-exchange HPLC of proteins using porous poly(glycidyl methacrylate-co ethylene dimethacrylate) monoliths grafted with poly(2-acrylamido-2-methyl-1 propanesulfonic acid). AB - Porous poly(glycidyl methacrylate-co-ethylene dimethacrylate) monoliths with different porous properties grafted with poly(2-acrylamido-2-methyl-1 propanesulfonic acid) chains using cerium(IV) initiated free-radical polymerization have been prepared and used for the separation of proteins in ion exchange HPLC mode. Because of the presence of the large pores that are typical of monolithic separation media which allow easy flow of all of the mobile phase, the efficiency of the columns does not deteriorate even at high flow velocities as a result of the specific morphology of the monoliths. Optimization of the chromatographic conditions such as the shape of the mobile phase gradient and the flow rate allows for very fast separation of three proteins in less than 1.5 min. PMID- 9336980 TI - Solvent and viscosity effects on the rate-limiting product release step of glucoamylase during maltose hydrolysis. AB - Release of product from the active site is the rate-limiting step in a number of enzymatic reactions, including maltose hydrolysis by glucoamylase (GA). With GA, an enzymatic conformational change has been associated with the product release step. Solvent characteristics such as viscosity can strongly influence protein conformational changes. Here we show that the rate-limiting step of GA has a rather complex dependence on solvent characteristics. Seven different cosolvents were added to the GA/maltose reaction solution. Five of the cosolvents, all having an ethylene glycol base, resulted in an increase in activity at low concentration of cosolvent and variable decreases in activity at higher concentrations. The increase in enzyme activity was dependent on polymer length of the cosolvent; the longer the polymer, the lower the concentration needed. The maximum increase in catalytic activity at 45 degrees C (40-45%) was obtained with the three longest polymers (degree of polymerization from 200 to 8000). A further increase in activity to 60-65% was obtained at 60 degrees C. The linear relationship between ln(kcat) and (viscosity)2 obtained with all the cosolvents provides further evidence that product release is the rate-limiting step in the GA catalytic mechanism. A substantial increase in the turnover rate of GA by addition of relatively small amounts of a cosolvent has potential applications for the food industry where high-fructose corn syrup (HFCS) is one of the primary products produced with GA. Since maltodextrin hydrolysis by GA is by far the slowest step in the production of HFCS, increasing the catalytic rate of GA can substantially reduce the process time. PMID- 9336981 TI - Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes. AB - The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and poly(vinylpyrrolidone) (molecular weight 40,000, PVP). Among the solutes examined, trehalose and sucrose protected the enzyme most effectively during storage at 37 and 45 degrees C. The restriction enzyme dried with trehalose or sucrose maintained its activity without detectable loss for at least 20 days at 37 degrees C and 12 days at 45 degrees C. In contrast, the activity of the enzyme dried with maltodextrin or PVP was reduced during vacuum desiccation and also it decreased remarkably during storage at the same temperatures. Stored (37/45 degrees C) vacuum-dried trehalose and sucrose systems were either a dense paste or a very viscous syrup, and this indicated that they were not glassy. Moreover, no relationship was found between the glass transition temperatures (Tg) of the pure added solute and enzyme protection during storage, since, e.g., sucrose which has significantly lower Tg values protected the enzyme much better than either maltose, lactose, maltodextrin, or PVP. The trisaccharide raffinose offered good protection of enzyme activity, and its role as a novel excipient matrix for labile enzyme stabilization deserves further investigation. The stability of enzyme EcoRI was rapidly lost when the vacuum-dried trehalose and sucrose systems were humidified to 58% relative humidity and stored at 45 degrees C, and this was attributed to disaccharide crystallization. PMID- 9336982 TI - Analysis of surface microtopography of biodegradable polymer matrices using confocal reflection microscopy. AB - Currently, synthetic degradable polymers are frequently employed as culture substrates prior to cell transplantation and as implantable scaffolds for cellular infiltration during soft and hard tissue repair. The surface microstructure of matrices based on such polymers may be important in controlling cellular anchorage, spreading, and growth on the external surface, as well as infiltration into the voids of porous polymer scaffolds. While the chemistry, bulk structure, and mechanical properties of such polymers have been extensively studied, the surface microstructure has not yet been systematically examined, particularly following polymer degradation. In this study, we present the first account of the use of confocal laser-scanning reflection microscopy (CLSM) to visualize and quantitate the microtopography of the surface of porous matrices of poly(lactic acid)/poly(glycolic acid) (PLAGA) copolymers following polymer degradation. Utilizing this technique, we report that the surface morphology of PLAGA matrices changes significantly upon degradation, with increased local clustering of textured regions. Our quantitative analysis suggests that polymer degradation results in a lower spatially-averaged surface roughness, with significant cyclical variations observed at later time points. The computed surface correlation function was observed to increase upon degradation, confirming the results from our morphological studies. Finally, we demonstrate the efficacy of CLSM to concomitantly image both the polymer surface and locally attached cells, in real time. PMID- 9336983 TI - Neutron and X-ray reflectivity studies of human serum albumin adsorption onto functionalized surfaces of self-assembled monolayers. AB - Neutron and X-ray reflectivity (NR and XR) have been widely used for the investigation of the structure of thin organic films. Here we demonstrate how these sensitive techniques may be applied to the study of protein adsorption to well-characterized self-assembled monolayers (SAMs) with different chemical functionalities. NR can be used for in situ study, while XR provides complementary information on the initial surfaces and dried layers measured in air after protein has been adsorbed. In situ measurements of adsorption of human serum albumin onto a hydrophilic NH2-terminated monolayer clearly show the presence of a thin layer of adsorbed protein next to the SAM. Adsorption of albumin onto a hydrophobic, deuterated, CD3-terminated SAM causes even bigger changes in the NR. Upon replacing the protein solution with protein-free buffer solution, the reflectivities from both kinds of monolayers do not change, demonstrating that the albumin adsorption is irreversible after several hours of contact with the protein solution. X-ray reflectivity measurements of dried substrates performed ex situ in air provide a lower bound estimate of the amount of protein which must be at the interface in situ. This combination of techniques provides a uniquely sensitive approach for studying changes that occur upon protein adsorption at an interface. PMID- 9336984 TI - Application of near-IR time-resolved spectroscopy and frequency response analysis for deep vein thrombosis detection. AB - Frequency response analysis is applied to analyze NIR-TRS spectra in a tissue model with a simulated thrombus. The value changes in parameters obtained from frequency response analysis are correlated with heterogeneity position in three dimensions. The goal of this research is to noninvasively localize deep vein thrombosis in the human leg through the use of this novel combination. PMID- 9336985 TI - Tumor detection and visualization using cyanine fluorochrome-labeled antibodies. AB - Tumor localization using fluorescence has been made practical by current improvements in tumor targeting molecules, especially monoclonal antibodies and their derivatives, by the development of convenient near-infrared emitting fluorochromes and by the availability of digital cameras having high sensitivity in this spectral region. Recent studies in animals have demonstrated that fluorochrome labeling of monoclonal antibodies confers adequate sensitivity and improved resolution. Distribution and catabolism of fluorochrome-labeled and radiolabeled antibodies are similar. Simultaneous localization of multiple reagents is made possible by labeling with several different near-infrared emitting fluorochromes; thus background subtraction and differential labeling of multiple tumor-associated components can be performed. Difficulties in using the fluorochrome labels are mainly related to light scattering and absorption in tissues, but detection of small tumors at depths of several millimeters is feasible. The major medical use of this new technology is likely to be endoscopic location of tumors. Scientific uses include studies of tumor metastasis, uptake and distribution of drugs and tumor-targeting molecules by tumors, and migration patterns of near-infrared labeled cells in vivo. PMID- 9336986 TI - Capillary electrophoresis in biotechnology. AB - Capillary electrophoresis (CE) is a versatile micro/macroanalytical technique gaining widespread usage for the separation and analysis of ionic substances. It has captured the attention of those working in a variety of arenas focused on biologically-active molecules. Its appealing characteristics include unprecedented mass sensitivity and the ability for precise, automated electrophoretic separation of microvolume samples with relatively short analysis times. Such versatility in bioanalysis makes it an inviting replacement for some of the labor-intensive, time-consuming methodologies performed via electrophoretic gels. Moreover, CE compliments the ease and speed of HPLC while eliminating the problem of excessive solvent volume usage and hazardous waste disposal. Further attractive characteristics of this technology include the analyses of a diverse spectrum of analytes, ranging from small organic ions to macromolecular protein complexes and DNA. While combining some of the most robust aspects of traditional electrophoresis, chromatography, and capillary technology, recent CE research and development has focused on avenues leading to improving detection and understanding and employing the basic chemistry of CE vis-a-vis new applications. PMID- 9336987 TI - Multipixel techniques for frequency-domain photon migration imaging. AB - The ability to map interior optical properties of a highly scattering medium from exterior measurements of light propagation is afforded by optical tomography. In this communication, we describe the problem of optical tomography, the techniques of photon migration measurements necessary to accomplish it, and the development of multipixel measurements for rapid collection of optical signals. These multipixel measurements are shown to provide detection of contrast owing to the optical properties of absorption and fluorescence associated with dye-laden heterogeneities embedded in a tissue-like scattering medium. From these rapid measurements, successful reconstruction of an interior optical property map may now be possible with clinically realistic data acquisition times. Applications for the technology arise for biomedical optical imaging for the in vivo detection of disease and the diagnosis of tissue (bio-) chemistry. PMID- 9336988 TI - Application of ultrasonic backscattering for level measurement and process monitoring of expanded-bed adsorption columns. AB - Expanded-bed adsorption is a newly commercialized technique for the purification of proteins from cellular debris in downstream processing. An expanded bed presents the possibility of protein recovery in a single step, eliminating the often costly clarification processing steps such as ultrafiltration, centrifugation, and precipitation. A major obstacle to the successful commercialization of this technology is the inability to accurately monitor and control the bed height in these systems. Fluctuations in the feedstock viscosity are common during normal operation and tend to make the operation and control of expanded beds for biological applications complex and difficult. We develop a level measurement technique based upon ultrasonics. It is shown that this technique has great promise for bed-height measurement in expanded-bed adsorption systems. Furthermore, the bed-height measurement can be used in feedback control strategies for bed-height regulation. The proposed ultrasonic sensor is also capable of monitoring for plugging and bubbling in the column. PMID- 9336989 TI - Effects of ammonium and lactate on growth and metabolism of a recombinant Chinese hamster ovary cell culture. AB - A Chinese hamster ovary (CHO) cell line producing a recombinant glycoprotein was cultured in batch mode with different initial concentrations of ammonium chloride (0-10 mM), sodium lactate (0-60 mM), or sodium chloride (0-60 mM). High ammonium concentrations did not inhibit cell growth and productivity or glucose and glutamine consumption. In contrast, specific ammonia and alanine production decreased by 55% and 40%, respectively. There were also significant increases in specific aspartate and glutamate consumption in high ammonium concentrations. These observations indicated a shift in glutamine catabolic pathways in response to the effects of ammonium. The influence of lactate on growth and metabolism were the combined effects of lactate concentration and osmolarity. After "correcting" for osmolarity effects, lactate was found to inhibit growth by 25% but to increase specific productivity slightly (10%). Lactate had profound effects not only on glycolysis but also on glutaminolysis. While specific glucose and glutamine consumptions decreased by 15-20%, the effects of lactate on their metabolic products were far more significant. Lactate production was halted, and specific ammonia and alanine productions decreased by 64% and 70% at high lactate concentration. Theories on how ammonium and lactate affected the metabolic pathways of glucose and glutamine are presented. PMID- 9336990 TI - Differences in sequence-specific expression of two anti-arsonate Fabs in E. coli. AB - Monoclonal antibodies are potentially useful therapeutic agents and can now be produced in hosts such as bacteria. However, it has been found that bacterial expression of some antibody-combining site fragments is greatly diminished. We compared two homologous anti-arsonate antibodies, 36-65 and 36-71, to address the question of why the former but not the latter expresses well as Fab in E. coli. These antibodies are both derived from the same variable region germline genes but differ in affinity due to somatic mutations present in 36-71. To investigate the poor expression of 36-71 Fab, we examined several factors, such as cellular toxicity, induction with isopropylthio-beta-D-galactoside, and growth of transformed bacteria at lower temperatures (30 degrees C), as well as the possibility of E. coli strain-related expression of Fab. However, none of these factors made a significant difference to Fab expression. We next localized a significant portion of the defect in Fab expression to the heavy chain by swapping the heavy and light chains from the two antibodies to construct hybrid Fabs. We used site-directed mutagenesis to engineer amino acids into the variable regions of antibody 36-71, to reproduce those found in 36-65 which is expressed well in E. coli. The defect in expression is due to residues located in the complementarity-determining regions, as mutations of heavy chain framework residues to those present in 36-65 do not enhance expression of 36-71 Fab in E. coli. PMID- 9336991 TI - Managed care and recovery: opportunities and challenges for psychiatric nursing. AB - The current crisis in mental health services is related to the "bottom line" focus of managed care. This trend as well as the consumer-led recovery movement challenge psychiatric nurses in both educational and clinical settings to carefully examine their knowledge, skills, and attitudes and to develop new partnerships with consumers in nontraditional settings to promote quality. The development of consumer-run peer support services serves as a model of consumer/professional partnerships. PMID- 9336992 TI - Characteristics of clients with schizophrenia who express certainty or uncertainty about continuing treatment with depot neuroleptic medication. AB - In this exploratory study, the factors corresponding to clients' certainty or uncertainty in continuing with depot neuroleptic medication for schizophrenia were examined. Ninety-four participants from a tertiary care schizophrenia clinic received an educational intervention aid containing information about treatment risks and benefits and were interviewed to elicit their levels of decisional conflict, self-efficacy, and emotional support, as well as expectations of risks and benefits of treatment. Eighty-seven percent of participants decided to continue treatment. Clients who expressed uncertainty (10%) about continuing with treatment had higher levels of decisional conflict (p = .000), lower levels of decision self-efficacy (p = .037) and decision emotional control (p = .003), lower expectations of hospitalization if treatment was stopped (p = .04) as well as lower expectations of benefits and higher expectations of side effects (p = .04), if treatment continued. In addition, reasons identified by participants for certainty or uncertainty were reported. The research may be useful in increasing the awareness of clinicians about the factors contributing to treatment decision making by clients diagnosed with schizophrenia and ultimately improve collaborative decision making. PMID- 9336993 TI - Communicating with individuals with Alzheimer's disease: examination of recommended strategies. AB - Meaningful conversation with individuals in the later stages of Alzheimer's disease (AD) has been considered difficult if not impossible. Limiting communication to simple concrete subjects and closed-ended questions is frequently recommended. Thirty-five 30 minute conversations with individuals with advanced AD (mean Mini-Mental State Examination [MMSE] = 10) were transcribed and the interactions examined. No significant differences in length or relevance of response by type of question was found indicating that subjects were able to respond to open-ended questions. Use of broad opening statements or questions, establishing commonalities, speaking as equals, and sharing of self-facilitated expression of feeling; recognizing themes with salience for the individual helped to maintain the discussion. PMID- 9336994 TI - Nurses' views regarding False Memory Syndrome. AB - Knowledge concerning the storage and retrieval of traumatic memories and so called False Memory Syndrome has not been widely available in nursing journals. Information in the popular media, however, means that nurses are learning about aspects of the memory debate from such sources. This article reports on 1,701 nurses' views of False Memory Syndrome (FMS). As background, this report reviews briefly current issues and research on traumatic memory retrieval. The majority of participants believed that FMS, although rare, could occur. For these nurses, FMS was a consequence of incompetent and unethical therapists. They worried that attention to FMS would silence or revictimize survivors of abuse. PMID- 9336995 TI - Body weight and shape self-cognitions, emotional distress, and disordered eating in middle adolescent girls. AB - In this study, stability of the body weight/shape self-schema and possible self in a sample of middle adolescent girls during their transition from junior high to high school was examined and the relationship between these self-cognitions and emotional distress and disordered eating behaviors was explored. Subjects (N = 79) completed measures of self-cognitions, competence, and self-esteem in the 8th and 9th grades. Disordered eating and depression were measured in 9th grade. Eighth grade self-schema scores were used to identify the fat/out-of-shape (n = 21) and thin/athletic (n = 20) self-schema groups. For both groups, stability in the body weight/shape cognitions was found. Girls in the fat/out-of-shape group had lower self-esteem, appearance, and athletic competence scores in both grades and higher dieting and depression scores in 9th grade than the slim/athletic group. Regression analyses showed that the self-schema score was a stronger predictor of the outcomes than weight. Findings suggest that the body weight/shape self-schema plays an important role in adolescent girls' emotional health. PMID- 9336996 TI - Psychometric analysis of the self-coherence survey. AB - Self-coherence, as measured by the Self-Coherence Survey, is an important determinant of psychosocial health in a variety of life circumstances. This report presents a psychometric analysis of the Self-Coherence Survey that resulted in three scales labelled Appraisal, Holism, and Introspection. The factorial structure, reliability, and validity of Self-Coherence was evaluated and replicated in two samples from the same community population (Phase I n = 433; Phase II n = 421). The three dimensions identified in the theoretical discussion of self-coherence published in this journal in 1993, were identified and replicated. Implications for further research and nursing practice are discussed. PMID- 9336997 TI - Divorcing and building a new life. AB - The purpose of this study was to provide a qualitative description of women's experiences of divorcing and building a new life. Interviews with 10 divorced women were conducted, transcribed, and analyzed using the constant comparative method. Four phases were identified in the process of divorcing and building a new life: the emotional divorce, making the decision, pulling apart, and moving beyond. Feelings and coping strategies reported by the participants are described. PMID- 9336998 TI - Cost effective use of laboratory tests in rheumatology. PMID- 9336999 TI - It is lupus? AB - It has been said that the essence of medicine is the reduction of uncertainty for patients and physicians (Ludo Baghius). Confronting patients who have some symptoms suggesting lupus but who do not meet criteria for classic SLE is challenging. Evaluating and caring for these patients, when not making a diagnosis of SLE, demands clinical acumen, confidence, and effective communication skills. PMID- 9337000 TI - Plasmodium falciparum malaria. PMID- 9337001 TI - Chloroquine versus pyrimethamine/sulphadoxine in the treatment of uncomplicated P. falciparum malaria in northern Kenya. AB - The response of P. Falciparum to chloroquine (CQ) and pyrimethamine-sulphadoxine (PSD) in vivo was investigated in 173 indigenous uncomplicated malaria patients at Sololo Hospital, Moyale district in northern Kenya. All the patients were symptomatic and parasitaemic. They were divided in two age groups (children < 10 years, adults > 10 years). They were randomly assigned to receive either CQ or PSD standard treatment and then followed up at 7 and 14 days. In the child group, out of 91 patients enrolled, 65 (71.4%) completed the seven-day study; among these 38 (17 females and 21 males with mean age of 41.9 months) were treated with CQ and 27 (11 females and 16 males with mean age 39.1 months) with PSD. Parasites were significantly (p < 0.001) more resistant to CQ (18/38, 47.4%) than PSD (0/27, 0%). In the adult group, out of 82 patients enrolled, 54 (65.9%) completed the 7-day-study, and among these 27 (10 females and 17 males with mean age of 22.5 years) were treated with CQ and 27 (11 females and 16 males with mean age of 23.2 years) with PSD. Parasites were significantly (p = 0.01) more resistant to CQ (7/20, 25.9%) than PSD (0/27, 0%). Overall, considering the 119 patients who completed the follow-up, the resistance of P. falciparum was significantly higher (p < 0.001) to CQ (25/65, 38.5%) than to PSD (0/54, 0%). Out of the 94 patients with negative slide at day 7, fifty seven came at the control of the day 14 (30 children and 27 adults). Among them, 22 were in CQ group and five were found positive (22.7%), while the 35 patients in PSD group all tested negative (p = 0.006). The resistance to CQ in the children group was 25% (p = 0.05) and 20% in the adult group (p = 0.13). We conclude that the significant parasitological resistance to CQ in the area under study questions the continued use of CQ as first line antimalarial treatment. On the contrary, PSD can still be considered a very effective drug against P. falciparum in northern Kenya. PMID- 9337002 TI - Effect of previous chloroquine intake on in vivo P. falciparum drug sensitivity. AB - Investigation of the in vivo response of P. falciparum malaria parasites to chloroquine was conducted during 1993/94 in Al-Ain District of Abu Dhabi Emirate, UAE. Sixty seven expatriates who developed falciparum malaria on their return from Pakistan, Oman and Sudan were recruited for the WHO in vivo tests. Of the 67 patients, eight were classified as having RII and RIII responses, while 59 remained aparasitaemic at the end of the seven-day WHO standard test. On continuation into the 28-day WHO extended test, a further 34 patients exhibited RI resistance. Resistance of parasites to chloroquine was confirmed by measurement of plasma chloroquine using High-Performance Liquid Chromatography. In all 67 patients, the level of chloroquine was well above the minimum therapeutic level. The outcome of the in vivo test in patients treated for the first time was significantly different from that in patients who were previously on chloroquine. Among patients treated for the first time, 36 out of 41 (88%) had a resistant response, whereas, among those previously on chloroquine only six out of 26 (23%) had a resistant response. The difference is probably due to the higher initial plasma level of chloroquine among patients who were previously on the drug. Curing more patients with higher plasma chloroquine implies that chloroquine shall continue to be useful, particularly if resistance is at the RI level. Appropriate higher therapeutic levels of chloroquine should be defined for such patients. PMID- 9337003 TI - Treatment of chloroquine-resistant malaria using pyrimethamine in combination with berberine, tetracycline or cotrimoxazole. AB - Two hundred and fifteen patients with chloroquine-resistant malaria were randomised into three groups. The first group of 82 patients were given pyrimethamine and berberine (berberine group), the second group of 64 patients, pyrimethamine and tetracycline (tetracycline group) and the third group of 69 patients were given pyrimethamine and cotrimoxazole (cotrimoxazole group). In the berberine group, the clearance, rate of asexual parasitaemia was 74.4% after treatment, while in the tetracycline group it was 67.2% and in the cotrimoxazole group 47.8%. These results indicate that berberine is more effective in clearing the parasite than both tetracycline and cotrimoxazole, and that the combination of pyrimethamine and berberine gives the best results for chloroquine resistant malaria. PMID- 9337004 TI - Retinal haemorrhage in cerebral malaria. AB - We examined the fundi of 73 children aged between six months and six years with confirmed diagnosis of cerebral malaria at the Children's Emergency Ward of the University College Hospital, Ibadan. Normal fundi, papilloedema and retinal haemorrhages were present in 38(52.1%), 18(24.7%) and 17(23.3%), respectively on admission. There were no significant differences between the three groups with respect to age, sex, admission coma score, posture, packed cell volume, parasite density, serum glucose, and serum electrolyte profile on admission. The mortality rates were 16%, 22% and 47% in the normal, papilloedema and retinal haemorrhage groups, respectively (Chi-squared for linear trend = 5.587, p = 0.018). Retinal haemorrhage was significantly associated with death (chi 2 = 5.846, p = 0.0192; Crude Odds ratio = 4.1, 95% CI = 1.1, 15.6; p = 0.018). The association was still present after adjusting for other known risk factors for mortality, including age, sex, acidosis, parasite density, anaemia, deep coma, and hypoglycaemia (adjusted Odds Ratio = 4.6, p = 0.0688). Papilloedema alone was not associated with mortality when compared with normal fundi [Fischer's exact (p = 0.448)]. It is concluded that fundoscopic abnormalities are common in children with cerebral malaria, and that retinal haemorrhage is associated with a poor prognosis in such children with cerebral malaria. Therefore, fundoscopic examination is not only useful to rule out raised intracranial pressure before performing a lumbar puncture, but also a useful measure in assessing prognosis in children suffering from cerebral malaria. PMID- 9337005 TI - Mosquito vectors of bancroftian filariasis in Kwale District, Kenya. AB - A total of 2,906 female mosquitoes were collected over a period of one year using pyrethrum spray-sheet and human bait methods, and dissected for filaria larvae in three hinterland villages of coastal Kenya. The dominant species, Anopheles gambiae and Anopheles funestus were also found to be the main vectors. From the spray catch collections 0,9 and 1 Cx. quinquefasciatus, An. gambiae and An. funestus out of 491, 708 and 403 respectively were infective. In the same order, 4, 2 and 2 out of 512, 196 and 180 from human bait collections were infective. The results indicate that Cx.quinquefasciatus is also an important vector in this area contrary to some previous findings that it played no important role in rural hinterland areas. Differences in the results from the human bait and spray catch methods have been pointed out and the advantage of using both methods in filarial surveys indicated. PMID- 9337006 TI - Parasitic infections in Pemba Island school children. AB - Intestinal helminths, schistosomiasis and malaria have been recognised for decades to be major public health problems in Zanzibar, Tanzania. During the evaluation of the impact of the Zanzibar Helminth Control Programme, baseline parasitological data on 3,605 school children were collected in Pemba Island. Prevalence of intestinal helminth infections was 72%, 94% and 96% for Ascaris lumbricoides, Trichuris trichiura and hookworm, respectively. Thirty one percent of children tested positive for haematuria, a reliable indicator of urinary schistosomiasis in the study area. Malaria parasites were found in 61% of children. Hookworm infections and haematuria were more prevalent in boys. Sixty seven percent of the children were infected with all the three helminths, and 28% harboured double infection. No association was found between intestinal helminths and schistosomiasis or malaria. Children living in rural areas were more heavily infected with hookworms, schistosomiasis and malaria compared to children in towns. Results from this study provided relevant information for designing a "plan of action" for the integrated control of filariasis, intestinal helminths, malaria and schistosomiasis in Zanzibar. PMID- 9337007 TI - Prevalence of adult diabetes in Ibadan, Nigeria. AB - Within the framework of an international collaborative network, we measured the prevalence of diabetes mellitus in a traditional Yoruba community in the city of Ibadan, Nigeria. Using a random sampling technique we enrolled a community sample of 247 men and women. Fasting blood glucose (FBG) was measured at the community clinic from a fingerstick using the Companion2 Medisense blood glucose meter. The mean FBG was 4.7 mmol/L and 4.9 mmol/L for men and women, respectively. Using the 1985 WHO criteria, the prevalence of diabetes was 2.8%. There was no significant rise in FBG with age. Compared to the lowest quartile of the body mass index (BMI), there was about a 1.5 fold increased risk of developing elevated FBG. The test of trend between FBG and BMI was however not statistically significant. Despite a modest hypertension rate (22.3%), there was no significant difference in the FBG for hypertensives compared to normotensives. The findings of this study show that the prevalence of diabetes mellitus in this West African community remains low compared to nations in Western societies. However, in comparison to previous estimates from sub-Saharan Africa, the prevalence of adult onset diabetes seems to be on the increase. PMID- 9337008 TI - Cardiovascular risk factors in middle aged Nigerians. AB - Ischaemic heart disease (IHD) contributes very little to mortality figures in Nigerians. In this study of 146 middle aged Nigerians (110 males and 36 females) aged 50-54 years, the cardiovascular risk factors of smoking, alcohol ingestion, history of diabetes mellitus, obesity, hypertension and high serum total cholesterol were assessed seeking to confirm the postulated reason for the low prevalence rate of IHD. Prevalence of the risk factors in this random sample population in males and females were as follows respectively: cigarettes 0%; alcohol intake 5.4% in males, 2.8% in females; self reported diabetes 1.8%, 2.8%; obesity 21%, 28%; hypertension 16.4%, 25% and cholesterol > 200 mg/d 6.4%, 13.9%. None of the subjects smoked more than 10 cigarettes a day. Multiple risk factors occurred infrequently in individual subjects. Only five men (4.5%) exhibited two risk factors and only one (0.9%) exhibited three risk factors apart from the gender. Of particular relevance, mean total cholesterol was 148.7 mg/dL (SD +/- 37.9) for the entire group, 143.5 mg/d (SD +/- 36.0) for men, and 153.6 mg/dl (SD +/- 35.3) for women. In the whole group, mean total cholesterol was higher in the hypertensive than the normotensive subjects (P < 0.05). Compared to similar age population from Japan and the United States, the prevalence of the risk factors was generally low. Attention should be on controlling obesity, hypertension and the diet to keep the prevalence low. PMID- 9337009 TI - Carcinoma of the oesophagus in Ibadan. AB - This was a retrospective analysis of 177 histologically confirmed cases of oesophageal carcinoma seen in the University College Hospital, Ibadan, Nigeria over a period of 30 years. Oesophageal carcinoma constituted 0.6 per cent of all malignant neoplasms and 1.4 cases per 1000 surgical biopsies during the study period. Dysphagia and weight loss were the most common clinical manifestations. Ninety three patients presented within one year of onset of clinical symptoms. The peak age incidence occurred in the seventh decade of life. Sex distribution was equal. The middle third of the oesophagus was the most common location of the neoplasm and the vast majority (94.5%) were squamous cell carcinomas. Achalasia of the cardia and Barrett's oesophagus were not associated with oesophageal carcinoma in this study. Regional lymph nodes and lungs were the most common sites of metastasis. Surgical complications included mediastinitis and bronchopneumonia, both occurring within seven days postoperatively. Late clinical presentation and high postoperative mortality are responsible for the persistently poor prognosis of oesophageal carcinoma despite significant advances in the diagnosis and management of these neoplasms. PMID- 9337011 TI - HIV antibody testing in a teaching hospital: policy versus practice. AB - The cost of HIV antibody testing can be phenomenal. Thus many countries, Ghana included, have adopted policies to guide physicians in making judicious test requests. An analysis of compliance with this policy in a teaching hospital in Ghana, shows that 70% of physician requests meet the stated Ministry of Health guidelines. However, while 84.5% of all test requests which turned positive were within stated guidelines, 48.6% of those turning out negative were not indicated by the policy. The cost of HIV antibody testing could be minimised if clinicians operated within the stated guidelines which make considerations for judicious use of health resources. Compliance with policy also needs evaluating. PMID- 9337010 TI - Human intestinal parasites in primary school children in Kampala, Uganda. AB - A cross sectional survey on intestinal parasite infections was carried out in 5,313 pupils between the ages of ten and fifteen years in 98 primary schools in Kampala. The aim was to identify the types and distribution of intestinal parasites and to estimate the prevalence in school children. Trichuris trichiura (28%), Ascaris lumbricoides (17%) and hookworms (12.9%) were common infections among the children. Other less commonly found parasites were S.mansoni, Strongyloides stercolaris, Taenia sp, Enterobius vermicularis, Giardia lamblia, Entamoeba coli and E. histolytica. Refuse dumps are probably a significant source of transmission of intestinal helminthic infections in Kampala. PMID- 9337012 TI - Criteria for better detection of brucellosis in the Narok District of Kenya. AB - Monthly disease summary sheets from 1986-1992 of 60 dispensaries, clinics and hospitals in Narok district, Kenya were reviewed for the occurrence of brucellosis and other diseases with "flu-like symptoms". Diseases with these symptoms accounted for about 52% of the 1,037,875 cases reported for the time period. These were classified as malaria (79.3%), rheumatism (7.1%), PUO (2.4%), and brucellosis (0.8%). Brucellosis was diagnosed by a positive Rose Bengal (RB) test routinely conducted in seven out of the 60 health units. In these units, 55% of flu-like cases were classified as malaria and 21.2% as brucellosis. Individual case records of patients at four dispensaries using the RB test during 1991-92 were assessed for specific predictor symptoms. For 625 RB tested patients, a positive test result was associated with joint pain, headache, and the combinations of joint pain with headache and lameness with headache. A logistic regression model correctly predicted the RB test result in 62.3% of the time. For the 465 patients examined by the blood smear examination, identification of malaria parasites was associated with, headache, joint pain and combinations of emesis with pale mucous membranes. This regression model correctly predicted positive results 67.2% of the time. Both models indicate that selected clinical predictors represented significantly increased odds of being positive to the respective tests. However, for both diseases, clinical signs alone appear insufficient for reliable diagnosis and differentiation probably due to resemblance in symptomatology between these two and other diseases. PMID- 9337013 TI - Reduced prevalence of onchocerciasis in Uganda following either deforestation or vector control with DDT. AB - To determine the prevalence of onchocerciasis in western Uganda following deforestation and vector control, three foci were re-examined 20 years after previous surveys. In the Ruteete focus Simulium neavei had apparently disappeared and the prevalence of onchocerciasis declined in adults from about 70% in 1971 to a standardised prevalence of 12% in 1992. An increase of population density together with extended deforestation was assumed as cause of this strong reduction. In Bugoye, a S. damnosum s.l. focus, the standardised prevalence of microfilaria carriers declined from 62% in 1972 to 4.7% in 1992. Entomological data indicated the absence of man biting blackflies in the nineties. It can be suggested that the vector control using DDT performed during the seventies had lead to a change of the species composition from anthropophilic to non anthropophilic S. damnosum s.l. In the focus Kicheche environmental changes were insignificant, deforestation was not progressive and S. neavei was abundant. Here the standardised prevalence of microfilaria carriers was still high (61%). PMID- 9337014 TI - Reduced prevalence of onchocerciasis following mass treatment with ivermectin. AB - The effect of four years of cummulative annual treatment with ivermectin just before the fifth round of dosing with the drug was studied in six endemic communities where pretreatment data had been collected in 1992. Significant reductions in prevalence of microfilaridermics (PMF), skin microfilarial density (MFD) and community microfilarial load (CMFL) was recorded. Remarkable reductions in PMF were recorded in Gbodongi (53.0%), Ndanako (80.1%) and others. In all the communities, the infection was reduced to a hypo endemic status except for Gbodongi. This was corroborated by the increase in amicrofilaridermics. CMFL was reduced in Gbodongi by 64.2%, Lata (53.1%) and the least in Bongi (11.0%). Reduction in clinical manifestations were variable. PMID- 9337015 TI - Serum immunoglobins in Nigerians with urinary schistosomiasis. AB - Serum levels of immuno-globulin G, A, M, D and E were determined using single radial immuno-diffusion technique in 52 Nigerian primary school children with urinary schistosomiasis and 39 age and sex matched controls. The mean values of IgA, IgM and IgE in the school children with S. haematobium infection were significantly higher than in controls. These findings suggest that urinary schistosomiasis in these subjects was in the acute stage. PMID- 9337016 TI - Surgical management of lumbar disc prolapse in Addis Ababa. AB - Of 40 cases of lumbar disc prolapse, 30 involved the L4/5 disc. There was a preponderance of males, M:F ratio of 5.7:1. All 40 cases were unresponsive to medical treatment and underwent bilateral laminectomies and discectomies at the respective levels. The operative findings were; two free prolapses, 34 major disc bulges compressing the lumbar theca and nerve roots, and four lateral disc protrusions. The post operative course was uneventful except one case with wound dehiscence. The remainder were discharged upon improvement. PMID- 9337017 TI - Protrusion of ventriculo peritoneal shunt through the anus: report of two cases. AB - Two cases of spontaneous protrusion of the abdominal end of ventriculo peritoneal shunts through the anus are reported in two African children with hydrocephalus. In each case the protruding catheter was easily removed with successful outcome and without recourse to major abdominal surgery. Possible factors suggested that may predispose to this complication of ventriculoperitoneal shunting include a weak bowel musculature in myelomeningocele and the use of stiff peritoneal catheters. PMID- 9337018 TI - In vitro changes of hydroxyapatite coatings. AB - The stability and degradability of hydroxyapatite coatings on dental implants depends on the dissolution of the individual chemical phases. Hydroxyapatite coated dental implants exhibit a range of amorphous-phase content. Two tests were conducted to observe the course of coating degradation. The first test showed degradation of both crystalline and amorphous coatings by cracking and dissolution after immersion in Ringer's solution. Concomitant saturation of the implants in the solution modified the coated surface with precipitated crystalline apatite. A second test, intended to replicate the conditions of infection by decreasing pH, illustrated preferred dissolution of the amorphous phase, liberating crystalline segments. It is expected that morphologic changes could influence the rate of bone bonding and therefore could alter or control implant-tissue interactions. PMID- 9337019 TI - Reverse-torque failure of screw-shaped implants in baboons after 6 months of healing. AB - Mechanical testing of the implant-tissue interface has been the focus of numerous investigations concerning the anchorage capacity of implants. The purpose of this study was to measure reverse-torque failure after 6 months of healing for three different biomaterials in the posterior jaws of four adult female baboons. The animals had all of their posterior teeth surgically extracted and, following 10 weeks of healing, 7 implants were placed in each quadrant. The biomaterials included titanium plasma-sprayed surfaces, titanium-aluminum-vanadium surfaces (both 3.8 mm x 10 mm), and a commercially pure titanium surface (3.75 mm x 10 mm). After 6 months, torque data were collected using a counterclockwise computerized torque driver and were analyzed by repeated measures analysis of variance for differences related to biomaterial, jaw, and biomaterial/jaw. Post hoc Tukey Kramer analysis was also performed for within-group differences (alpha = .05 level). The biomaterial comparison revealed a significant difference between the titanium plasma-sprayed and the combined commercially pure titanium/titanium -aluminum-vanadium groups (analysis of variance, Tukey Kramer, P < .05). The jaw comparison showed no significant difference, although the data suggest that higher forces may be required for mandibular torsional failure. The biomaterial/jaw comparison revealed that jaw differences for the mean values of commercially pure titanium and titanium-aluminum-vanadium implants were greater than jaw differences for mean values of titanium plasma-sprayed implants, although these differences were not statistically significant. Because of the lack of correlation between single-cycle biomechanical tests and clinical performance, it is necessary to be selective in assigning usefulness to data of this type. PMID- 9337020 TI - Bone formation and reosseointegration in peri-implantitis defects following surgical implantation of rhBMP-2. AB - This study was designed to evaluate bone formation and reosseointegration following surgical implantation of recombinant human bone morphogenetic protein-2 (rhBMP-2) in peri-implantitis defects. Hydroxyapatite-coated dental implants were placed bilaterally in the mandibular and maxillary premolar area in four rhesus monkeys and were allowed to osseointegrate for 1 year. Cotton ligatures were then placed around the healing abutments, and plaque was allowed to accumulate for 11 months. Resulting circumferential peri-implantitis defects exhibited a large intrabony and horizontal component. At reconstructive surgery, peri-implantitis defects in contralateral jaw quadrants were randomly assigned to receive rhBMP-2 (0.43 mg/mL implant volume) in an absorbable collagen sponge carrier or a carrier control. The animals were sacrificed 4 months postsurgery, and block sections were prepared for histometric analysis. Summary statistics included means calculated per animal. Paired t tests were used to evaluate differences between experimental conditions (n = 4). Defect depth amounted to 3.4 +/- 0.9 mm and 3.2 +/- 0.9 mm for rhBMP-2 and control defects, respectively. Vertical bone gain in rhBMP-2 defects (2.6 +/- 1.2 mm) was significantly greater than in controls (0.8 +/- 0.8 mm; P < .01). Reosseointegration within the confines of the defect for rhBMP-2 defects (29.0 +/- 10.5%) differed significantly from that in the control (3.5 +/- 2.5%; P < .01). Reosseointegration within the extent of newly formed bone averaged 40.0 +/- 11.0% in rhBMP-2 defects as compared to 8.9 +/- 7.8% in the control (P < .01). Osseointegration in resident bone amounted to 69.5 +/- 6.9% and 72.6 +/- 8.0% for rhBMP-2 and control defects, respectively. There is significant evidence that rhBMP-2 has potential to promote bone formation and reosseointegration in advanced peri-implantitis defects in a demanding nonhuman primate model. PMID- 9337021 TI - Evaluation of three different dental implants in ligature-induced peri implantitis in the beagle dog. Part I. Clinical evaluation. AB - The purpose of this study was to clinically evaluate experimental peri-implant breakdown. Hydroxyapatite-coated, titanium plasma-sprayed, and machined titanium alloy surfaces were investigated. Eighty-four implants were placed in 14 beagle dogs. Pocket probing depths and clinical attachment level and mobility measurements were made. Dogs were sacrificed at 3 and 6 months. All experimental implants showed a significant loss in clinical attachment level (P < .05). Increased pocket probing depths for experimental implants occurred during the first 2 months, after which a plateau was reached. At the 3- and 6-month evaluation, pocket probing depths at experimental implants were significantly increased (P < .05). No differences among the three implant types were noted for clinical attachment levels and pocket probing depths. In general, greater mobility was found with the titanium-alloy implants than with hydroxyapatite coated and titanium plasma-sprayed implants (P < .025). In addition, mobility measurements were significantly greater for experimental titanium-alloy implants during the first 3 months (P < .05). Clinical attachment level measurements were most sensitive to peri-implant status. All implants were equally susceptible to ligature-induced peri-implant breakdown. Consequently, meticulous oral hygiene and regular maintenance care are prerequisites for successful implantology. PMID- 9337022 TI - Passive film growth on titanium alloys: physicochemical and biologic considerations. AB - The role of reactive oxygen derivatives (hydroxy peroxide, hydroxyl radical, and singlet oxygen) on the precipitation of inorganic and organic complexes onto the surface of titanium implant alloys is discussed in this review. In addition, the effect of possible implication of several biologic entities surrounding the implant on the implant-tissue interface constituents is described. Evidence from relevant studies suggests that local microenvironmental byproducts and factors associated with the inflammatory response resulting from the implant-induced tissue insult may enhance the expressivity of the inherent, clinically important property of titanium to form oxides. Growth of titanium oxide may be explained through several processes derived from biologic, thermodynamic, and electrochemical approaches. The models proposed to interpret this phenomenon are often contradictory, demonstrating inward or outward from the bulk material passive film growth, with increasing or self-limiting levels of oxide formation as a function of time. However, in vivo observations are consistent with aging induced thickening of the complexes precipitated on the implant material surface. This review attempts to clarify several critical issues pertaining to passive film formation and kinetics on titanium-alloy surfaces. PMID- 9337023 TI - The DIA anatomic abutment system and telescopic prostheses: a clinical report. AB - Implant-supported prostheses may benefit from the versatility of design and favorable appearance offered by telescopic restorations. This project investigated the effectiveness of 208 abutments designed and produced by Dental Imaging Associates and the 73 prostheses supported by them over a 2-year period. No complications were found in the single-tooth group, although the sample was small (n = 7). Only 8.17% of the 208 abutment screws became loose during the initial postloading period. Once retightened, 2.4% of the total loosened for the second time. Higher screw-loosening rates were found in maxillary restorations, while the inclusion of a distal cantilevered pontic produced a significant increase in maintenance requirements. PMID- 9337024 TI - Effect of allogeneic, freeze-dried, demineralized bone matrix on guided bone regeneration in supra-alveolar peri-implant defects in dogs. AB - This randomized, split-mouth design study evaluated the adjunctive effect of allogeneic, freeze-dried, demineralized bone matrix on guided bone regeneration in a critical-size, supra-alveolar, peri-implant defect model. Contralateral supra-alveolar peri-implant defects, 5 mm in height, each including two titanium implants, were surgically created in five beagle dogs. Demineralized bone matrix in autologous blood was placed over the implants in one randomly selected mandibular jaw quadrant. A space-making expanded-polytetrafluoroethylene membrane was used to provide guided bone regeneration bilaterally. Following a 16-week healing interval, tissue blocks were harvested and prepared for histometric analysis. Differences between experimental conditions (guided bone regeneration sites with and without demineralized bone) were evaluated using paired t tests (n = 4). Demineralized bone particles were discernible, with limited signs of resorption. The bone matrix particles appeared to be solidified within a dense connective tissue matrix and in close contact with the implants. Limited matrix remineralization was apparent adjacent to the alveolar crest. No statistically significant differences were found between experimental conditions for any parameter examined. Peri-implant defect height averaged 5.0 +/- 0.2 mm and 4.9 +/ 0.4 mm, vertical bone regeneration 1.5 +/- 0.9 mm and 1.1 +/- 0.4 mm, osseointegration within the extent of the defect 10.0 +/- 3.9% and 15.3 +/- 5.3%, osseointegration within the extent of regenerated bone 30.4 +/- 13.7% and 52.1 +/ 17.9%, and osseointegration within the alveolar base 68.8 +/- 13.1% and 74.4 +/- 7.1% for guided bone sites with and without demineralized bone, respectively (P > .05). The results suggest that freeze-dried demineralized bone has no adjunctive effect on guided bone regeneration in supra-alveolar peri-implant defects, that guided bone regeneration has a limited potential to enhance alveolar regeneration in this defect model, and that a 16-week healing interval appears insufficient for turnover and maturation of demineralized bone under provisions for guided bone regeneration. PMID- 9337025 TI - Dental implant survival in the irradiated jaw: a preliminary report. AB - Seventeen oral cancer patients (47 to 78 years, mean 67) were treated with external radiation in areas that included future implant sites. Implants were placed in the irradiated jaws after a period of 18 to 228 months (mean 88 months). The patients received 103 implants and were followed for 1 to 62 months (mean 21 months) after implant loading. The cumulative survival rate of the implants after 1 year was 97% in the mandible and 92% in the maxilla. While the irradiation dose used did not affect implant survival, this result may possible be influenced by the addition of hyperbaric oxygen treatment for patients receiving more than 50 Gy. Irradiation for treatment of oral cancer does not seem to reduce the survival rate of implants as compared to those placed in the nonirradiated jaw. PMID- 9337026 TI - Osteoblast responses to BMP-2-treated titanium in vitro. AB - In this study, the phenotypic expression of osteoblast progenitor cells, 2T9, on characterized titanium surfaces was examined in the presence of a bone morphogenetic protein-2 (BMP-2). X-ray photoelectron spectroscopy spectra indicated the presence of titanium dioxide on titanium surfaces, along with surface contaminants such as carbon and nitrogen. In the in vitro study, the activity of BMP-2-treated osteoblast cells on titanium surfaces was marked by significantly higher alkaline phosphatase-specific activity and 1,25 (OH2) vitamin D3-stimulated osteocalcin production when compared to the untreated cells on titanium surfaces. PMID- 9337027 TI - Mandibular fracture after endosseous implant placement in conjunction with inferior alveolar nerve transposition: a patient treatment report. AB - A patient with a severely atrophic right posterior mandible had three endosseous implants placed in conjunction with transposition of the inferior alveolar nerve. Three weeks following implant placement surgery, the patient experienced a spontaneous fracture of the mandible involving the two anterior implants. The two implants were removed, and the fracture was treated with open reduction and fixation with titanium mesh. The fracture healed, and the posterior implant integrated. This report suggests that the buccolingual and superior-inferior position of the mandibular canal can increase the possibility of mandibular fracture by increasing the size of the buccal cortical plate that is removed to expose the nerve during surgery. PMID- 9337028 TI - The effect of luting agents on the retention and marginal adaptation of the CeraOne implant system. AB - In this study, various luting agents were evaluated to determine their retentive strengths as they pertain to the CeraOne single-tooth implant system. Ten samples of five different luting agents (zinc oxide-eugenol, glass-ionomer cement, hybrid glass-ionomer cement, composite resin, and zinc phosphate) were tested for retentive strength of the CeraOne gold cylinder to the CeraOne abutment. Under the conditions of the experiment, zinc phosphate showed a mean retentive strength 164% greater than that of glass-ionomer cement and 49% greater than that of composite resin cement. Scanning electron micrographs were taken to evaluate the effect of various luting agents on marginal opening. The measurements revealed that zinc phosphate had the greatest marginal opening, although its mean value of 62 microns is within clinically acceptable limits. PMID- 9337029 TI - Cross-infection from periodontitis sites to failing implant sites in the same mouth. AB - Twenty-five tooth and implant sites in nine patients were investigated for the presence of putative periodontopathic organisms, using specific DNA probes for Actinobacillus actinomycetemcomitans, Porphromonas gingivalis, Prevotella intermedia, Eikenella corrodens, Fusobacterium nucleatum, Treponema denticola, and Campylobacter recta. Five of nine patients showed a likelihood of transmission from tooth to implant sites. These patients also showed a high number of putative periodontopathic organisms present in the tooth sites tested. A significant risk was found of transmitting putative periodontopathic organisms from periodontitis sites to implant sites in the same mouth. An appropriate clinical protocol needs to be developed to address this issue. PMID- 9337030 TI - Rigid connections between natural teeth and implants: a technical note. AB - In the posterior partially edentulous jaw, implants may be used to supplement existing natural dentition. Frequently, the maxillary sinuses and the mandibular nerve preclude the fabrication of freestanding implant-retained prostheses. However, if an implant and a natural abutment are combined, a fixed prosthesis can be fabricated, restoring the arch into the premolar area. The histories of three patients with attachments connecting implant-retained ceramotitanium crowns with crowns on natural abutments are described. A design for a rigid custom-made attachment for the Branemark system, using standard components with a machine duplication, spark-erosion technique, is suggested. PMID- 9337031 TI - Implant-supported prostheses in patients with Sjogren's syndrome: a clinical report on three patients. AB - Implant-supported prostheses offer a solution to the problems experienced by edentulous patients with Sjogren's syndrome. These patients often find it very difficult, if not impossible, to wear conventional complete dentures. Three clinical reports provide an insight into some of the difficulties involved in treating patients afflicted with this complex multifactorial disease using the Branemark system. PMID- 9337032 TI - Immediate labial contour restoration for improved esthetics: a radiographic study on bone splitting in anterior single-tooth replacement. AB - A bone-splitting technique used for anterior single-tooth replacement was evaluated in 54 patients and 68 sites. The cumulative rate of implant survival was 93.7% (SE 4.6%) after more than 4 years. The decrease in marginal bone height ranged from 0.8 to 1.3 mm. Some reaction of the bone levels around the adjacent teeth should be anticipated (0.3 to 0.5 mm). It was concluded that the bone splitting procedure is a safe and predictable technique when performed carefully on selected patients and with the proper instrumentation. The procedure seeks to reconstruct the labial contour of the alveolar process, which is a prerequisite for optimal and lasting implant esthetics. PMID- 9337053 TI - Enhancement of megakaryocytopoiesis by Campath-1G-treated natural killer cells. AB - Campath-1G is a CD52 (rat IgG2b) moAb used in bone marrow transplantation (BMT) to prevent graft-versus-host disease (GVHD) by the elimination of T cells via antibody-dependent cell cytotoxicity (ADCC) in vivo. We have previously reported that Campath-1G induces T cell proliferation, activation, and production of cytokines which in turn causes an enhancement of megakaryocytopoiesis in vitro. In view of the fact that recent studies have indicated that natural killer (NK) cells may also be involved in the regulation of megakaryocytopoiesis, we undertook the study of the in vitro effect of Campath-1G-treated NK cells on the regulation of megakaryocytopoiesis. Early burst-forming BFU-MK and late colony forming CFU-MK were grown from 2 x 10(5) peripheral blood non-adherent mononuclear cells (NAMC) in plasma clots in the presence of aplastic canine plasma (PICS-J) which was used as megakaryocyte colony-stimulating factor (MK CSF). The first step in elucidating this series of events was to investigate the direct influence of NK cells on megakaryocytopoiesis. Co-culturing NK cells (>85% CD16+) with autologous NAMC at a ratio of 1:1 resulted in a significant increase in the proliferation of CFU-MK and BFU-MK over NAMC cultured alone. This effect was further enhanced upon exposure of NK to Campath-1G (0.1-3 microg/ml). To investigate the possible influence of soluble factors released from NK cells treated with Campath-1G on MK maturation, conditioned medium (CM) derived from Campath-1G-treated-enriched populations of NK cells was found to enhance MK progenitor growth. Our data demonstrate that resting and Campath-1G-treated NK may be involved in the immunomodulation of megakaryocytopoiesis. PMID- 9337054 TI - Allogeneic peripheral blood progenitor cells for treatment of relapse after bone marrow transplantation. AB - Donor leukocyte infusions (DLI) are an effective therapy for patients who relapse with leukemia after bone marrow transplantation (BMT). Severe graft-versus-host disease and prolonged periods of pancytopenia compromise the success of this treatment in a substantial number of patients. We used filgrastim-mobilized peripheral blood progenitor cells (PBPCs), in some cases preceded by cytoreductive therapy, to circumvent some of the problems associated with DLI. Eleven patients (median age 41 years) received a total of 20 donor cell infusions. Their diagnosis was CML in hematological (two patients) or cytogenetic relapse (two patients), six patients suffered from acute myeloid leukemia (AM; n = 5) or Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL Ph+). One patient had multiple myeloma (MM). All six patients with acute leukemias received cytoreductive therapy prior to PBPC infusions; three patients with CML were pretreated with IFN alpha. Four of four patients with CML responded to PBPC infusions and currently are in complete clinical and molecular remission for time periods between 1 and 12 months. Six of six patients with acute leukemias achieved a complete remission. All of them relapsed after a median remission duration of 24 weeks (range 11-49 weeks). Three patients relapsed at extramedullary sites (CNS, testes, skin). Four of six acute leukemia patients received further cytoreductive therapy. All patients responded again and are in complete remission for time periods between 14 and 615 days. Two patients with acute leukemias have died due to dissemination of the disease. The patient with MM did not respond and is alive with disease. Severe (grade III) acute GVHD developed in two of 11 patients, three patients developed grade II disease, six patients did not show any signs of GVHD. Extensive chronic GVHD has developed in two cases to date. Patients with chemotherapy prior to PBPC infusion developed neutropenia and thrombocytopenia with a maximum duration of 20 and 14 days, respectively; prolonged periods of neutropenia did not occur. Two patients developed long-lasting thrombocytopenia in spite of PBPC infusion, in one case followed by leukemic relapse. Repeated courses of chemotherapy and PBPC infusion were generally tolerated well; no early deaths due to treatment-related toxicity or GVHD were observed. We conclude that the use of allogeneic PBPC instead of DLI in patients with relapse after BMT is technically feasible and safe. The efficacy of PBPC infusions seems comparable to DLI in patients with CML. Patients with acute leukemias also achieved complete albeit transient remissions. Aggressive chemotherapy followed by PBPC infusions resulted in only limited duration of cytopenia. The usage of PBPC infusion instead of non G-CSF-mobilized donor cells for treatment of relapse after BMT may reduce pancytopenia-related complications and merits further investigation. PMID- 9337055 TI - Treatment of poor-risk neuroblastoma patients with high-dose chemotherapy and autologous peripheral stem cell rescue. AB - A single institutional pilot study was conducted in which 12 poor-risk neuroblastoma (NB) patients were uniformly treated with multi-agent induction chemotherapy followed by myeloablative consolidation chemotherapy and unpurged peripheral blood stem cell (PBSC) rescue. In addition to using standard criteria for evaluating response to induction chemotherapy, tumor cell contamination of the peripheral blood and/or bone marrow was analyzed in seven patients by immunocytology using a panel of five anti-NB monoclonal antibodies. Seven patients had morphologic evidence of bone marrow disease at the time of diagnosis, and two additional patients had tumor cells detected in bone marrow samples by immunocytology prior to the second cycle of chemotherapy. After three cycles of chemotherapy, two of the 12 patients continued to have evidence of bone marrow disease. Samples from 29 PBSC harvests collected from nine patients were also analyzed for the presence of contaminating tumor cells by immunocytology. In each case, the stem cells were found to be free of tumor. Eleven of the 12 patients underwent myeloablative therapy and PBSC rescue; five patients remain alive without disease progression, 28+ to 53+ months from diagnosis, and six patients have developed recurrent disease. We conclude that PBSCs can be successfully harvested from children with NB, and used for hematopoietic reconstitution following myeloablative chemotherapy. However, more effective therapy for poor-risk NB patients is still urgently needed. PMID- 9337056 TI - Long-term results after allogeneic bone marrow transplantation for chronic myelogenous leukemia in chronic phase: a report from the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation. AB - The purpose of this study was to determine the long-term results of allogeneic bone marrow transplantation for chronic myeloid leukemia. A retrospective analysis was carried out of the outcome of 373 consecutive transplants performed at 38 European institutions between 1980 and 1988 and reported to the registry of the European Group for Blood and Marrow Transplantation. All transplants were carried out for first chronic phase of chronic myelogenous leukemia using unmanipulated marow cells from HLA-identical sibling donors. The probability of survival and leukemia-free survival at 8 years were 54% (95% CI: 49-59) and 47% (95% CI: 41-52) respectively. The probabilities of developing acute GVHD (II-IV) at 100 days and chronic GVHD at 4 years after transplant were 47% (95% CI: 41-53) and 52% (95% CI: 46-58) respectively. The probabilities of transplant-related mortality and leukemic relapse 8 years after BMT were 41% (95% CI: 36-48) and 19% (95% CI: 14-25), respectively. Transplant within 12 months of diagnosis was associated with reduced transplant-related mortality (34 vs 45%, P = 0.013) and resulted in improved leukemia-free survival (52 vs 44%, P = 0.03). The probability of relapse was significantly reduced in patients who developed chronic GVHD (RR = 0.33, P = 0.004). The probability of relapse occurring more than 2 years after transplant was increased more than five-fold in patients transplanted from a male donor (RR = 5.5, P = 0.006). Sixty-seven patients in hematologic remission were studied for residual disease by two-step RT/PCR for BCR-ABL mRNA and 61 (91%) tested negative. We conclude that bone marrow transplantation can induce long-term survival in approximately one-half of CML patients; the majority of survivors have no evidence of residual leukemia cells when studied by molecular techniques. The probability of late relapse is increased with use of a male donor. PMID- 9337057 TI - Impaired androgen production in female adolescents and young adults after total body irradiation prior to BMT in childhood. AB - Pubertal development and androgen production were evaluated 1-10 years after bone marrow transplantation (BMT) in 15 females aged 14-23 (mean 17) years. Before BMT, these patients had received combination chemotherapy for hematologic malignancy, and all had had a transplant program including total body irradiation (TBI). Of the nine patients who were pre-menarcheal at BMT, two had subsequently experienced spontaneous menarche at 11.5 and 13.3 years of age. Six were post menarcheal, but became amenorrheic after BMT. Menstruation subsequently started spontaneously in one of them 6 years after BMT. At the time of the study, three patients were early to mid-pubertal and 12 late pubertal or post-pubertal. Twelve patients were receiving sex steroid substitution therapy. Serum concentrations of testosterone, androstenedione, dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEAS) were determined. Androgen levels of late pubertal and post-pubertal transplanted patients were compared with 19 post-menarcheal patients aged 14-21 (mean 17) years who had been treated for hematologic malignancy with conventional chemotherapy. Testosterone levels of 52 healthy post-menarcheal females aged 14 29 (mean 19) years were measured as controls. Androgen levels of the BMT patients were lower than those of the conventionally treated patients. Differences in testosterone, androstenedione and DHEA levels were significant. Three spontaneously menstruating BMT patients had normal androgen levels. Testosterone levels of the conventionally treated patients and healthy controls were similar. Subnormal androgen production might be one factor behind the problems in pubertal development and sex life experienced by females after BMT. The use of these hormone levels for follow-up purposes and the potential value of androgen replacement therapy in females after TBI merit further study. PMID- 9337058 TI - Growth after recombinant human growth hormone (rhGH) treatment in transplanted thalassemic patients. AB - The aim of this study was to evaluate the treatment effects with recombinant human growth hormone (rhGH) in a group of patients after bone marrow transplantation for thalassemia major. At the end of treatment we divided the subjects into two groups according to the outcome of the therapy: responder and nonresponder. Responder group: after 24 months of rhGH administration, growth rate was still significantly higher in respect to start of treatment (P < 0.0001). Plasma levels of IGF-I rose significantly (P < 0.003). The serum levels of serum asparate aminotransferase (SGOT) and alanine aminotransferase (SGPT) were higher compared to normal values but improved in non-responder patients. There was no difference in the mean concentration of these parameters before and after treatment (P = NS). Non-responder group: these patients had a worsening of the growth rate during rhGH administration. There was no increase of the IGF-I levels. Single values of transaminase and ferritin levels were higher than in responder patients before and after treatment. There was a significant correlation between IGF-I, SGOT, SGPT and ferritin in all patients before and after therapy. It appears from these data that rhGH administration is worth serious consideration in patients after BMT for thalassemia major presenting impaired growth hormone secretion. This treatment can offer good results only in cases where the normal hepatic synthesis of IGF-I is conserved and where liver damage has not reached irreversible conditions, as we have seen in the responder group. PMID- 9337059 TI - Major salivary gland dysfunction in patients with hematological malignancies receiving interleukin-2-based immunotherapy post-autologous blood stem cell transplantation (ABSCT). AB - lnterleukin-2 (IL-2) is known to cause xerostomia and skin manifestations similar to graft-versus-host disease (GVHD). We therefore evaluated major salivary gland function in patients with hematological malignancies treated with IL-2 and interferon-alpha (IFN-alpha) after ABSCT. Eleven patients (seven male, four female) of median age 40 (24-47) were evaluated, seven with non-Hodgkin lymphoma (NHL); one with Hodgkin's disease (HD) and three with acute myelogenous leukemia (AML). Parotid and submandibular salivary gland function was assessed before, during and after IL-2/IFN-alpha administration by evaluation of the salivary flow rate and the composition of secreted saliva. Significant reductions in both the resting and stimulated parotid and submandibular salivary flow rates were observed during IL-2/IFN-alpha immunotherapy compared with the pre- and post therapy values (P < 0.01), while no hyposalivation was observed in the control patients who underwent ABSCT and did not received IL-2. Sialochemical evaluation revealed a significant increase in potassium concentration (24.4+/-0.6 mEq/l to 28.9+/-1.4 mEq/l) and a significant decrease in sodium concentration (6.7+/-2.1 mEq/l to 3.3+/-1.0 mEq/l) (P < 0.05) in the stimulated parotid gland saliva secreted during IL-2/IFN-alpha administration. Salivary protein concentrations were not altered by the IL-2/IFN-alpha immunotherapy. Similar changes were previously observed in mice and humans with chronic GVHD. We conclude that IL-2 immunotherapy induces major salivary gland dysfunction in humans, similar to our previous observations in patients with chronic GVHD, which may indicate similar pathophysiologic mechanisms. PMID- 9337060 TI - Value of cytomegalovirus detection by PCR in bronchoalveolar lavage routinely performed in asymptomatic bone marrow recipients. AB - In order to compare PCR with rapid virus culture for the early detection of CMV in bronchoalveolar lavage (BAL) after bone marrow transplantation, 26 asymptomatic patients were routinely evaluated for the presence of CMV on day 35 using these two techniques. Concurrent blood samples were also analyzed in all cases. CMV was detected synchronously by both culture and PCR in six of 26 (23%) BAL and in five of 26 (19%) blood specimens. Among these positive specimens, three BAL and blood samples were positive in the same patients. Five (19%) BAL and five (19%) blood samples were culture-negative but PCR-positive. No BAL or blood specimens were positive by culture alone. When considering matched BAL blood samples, five were positive in only one fluid, BAL (n = 3) or blood (n = 2) using culture, while seven were positive in only one fluid, BAL (n = 4) or blood (in = 3) using PCR. Overall, six of 26 (23%) patients had culture-negative but PCR-positive results. Three of these six patients were positive only in BAL and two of them subsequently received antiviral therapy for development of symptoms suggestive of CMV infection. We suggest that asymptomatic patients with negative culture but PCR-positive results on day 35 in BAL should be subsequently closely monitored for the presence of CMV. PMID- 9337061 TI - CD34 counts to predict the adequate collection of peripheral blood progenitor cells. AB - An essential prerequisite for successful procurement of sufficient autologous peripheral blood progenitor cells (PBPC) for engraftment is the optimal timing of collection. A number of surrogate markers of peripheral blood progenitor cells were analysed to identify a single test which could predict the optimum time to harvest, providing at least 2 x 10(6) CD34+ cells/kg patient body weight. The study comprised 95 patients undergoing varied mobilisation regimens with chemotherapy and G-CSF for both solid tumours and haematological malignancies. One hundred and fifty-seven PBPC harvests were collected. Full blood counts (FBC) and CD34+ cell enumeration was performed on blood samples taken during the mobilisation period and immediately prior to leucapheresis (pre-harvest). All PBPC collections were assayed for colony-forming cells and CD34+ cells in addition to a FBC. The white cell count on the day of harvest showed only weak correlation with the total number of CD34+ cells in the collection (r = 0.30). In contrast, the absolute number of circulating CD34+ cells strongly correlated with the CD34+ cell and CFU-GM yield of the corresponding apheresis product. Provided the mobilisation sample contained > or =20 x 10(6) CD34+ cells/ml, 94% of single collections, performed the following day, contained > or =2 x 10(6) CD34+ cells/kg. PMID- 9337062 TI - Expression of CD44 isoforms by highly enriched CD34-positive cells in cord blood, bone marrow and leukaphereses. AB - CD34-positive cells were isolated from cord blood (n = 8), bone marrow (n = 4) and leukapheresed material (n = 7), using an immunomagnetic isolation technique, MACS (Miltenyi Biotec, Bergisch Gladbach, Germany). In flow cytometric analysis, cell populations after enrichment revealed a fraction of 96.1% (cord blood), 96.2% (bone marrow) and 98.6% (leukapheresis material) CD34-positive cells. Cells were further stained with antibodies specific for CD44 isoforms: CD44s (SFF-2), CD44v5 (VFF-8) and CD44v6 (VFF-18). CD44-positive cells were detected by directly (FITC, fluorescein isothiocyanate) or indirectly (streptavidin-PE, phycoerythrin) conjugated fluorochromes in flow cytometric analysis. Analysis was restricted to CD34-positive cells. A high expression of CD44s was noted in all kinds of material under investigation with mean values in the range of 98.6-100%. There was little expression of CD44v6 (mean values in the range of 1.5-3.6%) and very slight expression of CD44v5 (mean values in the range of 0.6-1.4%). The finding that CD34-positive hematopoietic stem cells express CD44v5 and CD44v6 to a very small extent offers the possibility of using antibodies specific to CD44v5 and CD44v6 in immunopurging in the course of autologous stem cell transplantation. PMID- 9337064 TI - Celiac disease transmitted by allogeneic non-T cell-depleted bone marrow transplantation. AB - We observed the occurrence of celiac disease following allogeneic bone marrow transplantation in a patient transplanted for acute leukemia. The marrow donor was his HLA-identical sister, who had suffered from celiac disease since birth. The post-transplant period was characterized by recurrent episodes of diarrhea. Detailed workup showed atrophic intestinal mucosa on histology and anti-gliadin and anti-endomysium antibodies in the serum, features that were not present before transplantation. GVHD was absent at that time. The patient remains free of symptoms on gluten-free diet and slight immunosuppression. This case suggests transmission of celiac disease by bone marrow transplantation and supports the T cell concept in celiac disease. PMID- 9337063 TI - A laboratory comparison of T cell depletion by CD34+ cell immunoaffinity selection and in vitro Campath-1M treatment: clinical implications for bone marrow transplantation and donor leukocyte therapy. AB - Donor leukocyte infusions (DLI) have been used effectively to induce remission in patients who relapse after BMT. Using CD34+ cell immunoaffinity enrichment, donor T cells may be captured in the unadsorbed (residual) fraction and we assessed this as a potential source of functional T cells for post-BMT immunotherapy. We extended our study to compare CD34+ cell selection and antibody-mediated cell lysis using Campath-1M and measured T cell-depletion, CD34+ cell recovery and relative progenitor proliferative potential. The recovery of CD3+ cells (responsive to IL-2 or PHA) in the unadsorbed fraction was 84+/-12% (mean+/-s.d.) using a laboratory scale CD34+ cell selection process (CEPRATE LC). The immunoselected (CD34+ cell enriched) product contained 55+/-12% of the starting CD34+ cells (purity, 75+/-6%) with recoveries of 44+/-12% and 42+/-13% for CFU-GM and BFU-E respectively. T cell depletion was 99.8+/-0.2% (FACS) and the frequency of clonable T cells estimated at 1:640 (limiting dilution assay). In comparison, Campath-1M-treated marrow samples gave recoveries of CD34+ cells, CFU-GM and BFU E of 50+/-7%, 78+/-20% and 79+/-18%, respectively. The frequency of clonable T cells was 1:2700 despite an estimated T cell depletion of 98.4+/-1.9%. Data obtained from four BM harvests processed on the clinical grade CEPRATE SC system was comparable in every respect to the laboratory scale system. The yield of 1259 +/- 222 x 10(6) CD3+ cells in the unadsorbed fraction would allow for multiple graded incremental T cell aliquots for DLI for patients with acute leukaemia. PMID- 9337065 TI - Autologous stem cell infusion for acute myeloblastic leukemia in an HIV-1 carrier. AB - We present the case of an asymptomatic HIV carrier, who presented with acute myeloblastic leukemia in third relapse and successfully underwent autologous stem cell transplantation as a rescue treatment. This observation supports the conclusion that tolerance of autologous bone marrow or stem cell transplant in patients with HIV may correlate with a low viral burden and relatively good immune function. PMID- 9337066 TI - Autologous transplantation of G-CSF mobilized bone marrow cells in a child with disseminated fibrosarcoma. AB - We report a very rapid engraftment after reinfusion of bone marrow cells derived from 'G-CSF-primed bone marrow' in a small child. As the hemopoietic recovery was equal to that seen after PBSCT, we suggest that the use of 'mobilized bone marrow' is a good alternative for PBSCT in children if contraindications to the collection of PBSCT are present. PMID- 9337067 TI - Complete cytogenetic response with host-derived hematopoiesis induced by cyclosporin A discontinuation in a patient with relapsed chronic myelogenous leukemia after bone marrow transplantation. AB - A 46-year-old woman with Ph-positive CML received an unmanipulated BMT from an HLA-identical brother, conditioned with busulfan-cyclophosphamide. Five months after BMT, cytogenetic relapse occurred, and CsA was decreased and then discontinued. Mild acute GVHD occurred, but gradually improved with no immunosuppression. Forty days after CsA discontinuation, both cytogenetic and fluorescence in situ hybridization analyses showed a host-derived normal karyotype, 46,XX, and no evidence of leukemic cells or donor graft. The sustained host-derived hematopoiesis lasted for 2 years until sudden recurrence of CML. In this case, the discontinuation of CsA led to GVHD and also suppression of the relapsed leukemia, presumably by a 'specific' GVL effect. There was also graft failure. The observation that subsequent hematologic recovery was of host origin implies that, at least in this case, the GVL effect was not directed against normal host-type hematopoiesis. PMID- 9337068 TI - Reactive oxygen species and Alzheimer's disease. AB - Although a consensus that Alzheimer's disease (AD) is a single disease has not been reached yet, the involvement of the amyloid precursor protein (APP) and betaA4 (A beta) in the pathologic changes advances our understanding of the underlying molecular alterations. Increasing evidence implicates oxidative stress in the neurodegenerative process of AD. This hypothesis is based on the toxicity of betaA4 in cell cultures, and the findings that aggregation of betaA4 can be induced by metal-catalyzed oxidation and that free oxygen radicals may be involved in APP metabolism. Another neurological disorder, familial amyotrophic lateral sclerosis (FALS), supports our view that AD and FALS may be linked through a common mechanism. In FALS, SOD-Cu(I) complexes are affected by hydrogen peroxide and free radicals are produced. In AD, the reduction of Cu(II) to Cu(I) by APP involves an electron-transfer reaction and could also lead to a production of hydroxyl radicals. Thus, copper-mediated toxicity of APP-Cu(II)/(I) complexes may contribute to neurodegeneration in AD. PMID- 9337069 TI - Human ovarian cancer of the surface epithelium. AB - Epidemiologic studies have shown that the risk of cancer in the ovarian surface epithelium is decreased by factors that suppress ovulation, whereas uninterrupted ovulation has been associated with increased risk. This suggests that ovulation may play a critical role in ovarian carcinogenesis. More recently, molecular studies have demonstrated alterations in specific oncogenes and tumor suppressor genes in ovarian cancers. Overexpression of the HER-2/neu oncogene occurs in approximately 30% of ovarian cancers and correlates with poor survival. Although mutation of the K-ras oncogene has been found in some mucinous ovarian cancers, mutations in this gene appear to be more common in borderline ovarian tumors. Amplification of c-myc occurs in approximately 30% of ovarian cancers and is more frequently seen in serous cancers. Mutation of the p53 tumor suppressor gene, with resultant overexpression of mutant p53 protein, occurs in 50% of stage III/IV and 15% of stage I/II ovarian cancers. Most p53 mutations in ovarian cancers are transitions, which suggests that they arise spontaneously rather than due to exogenous carcinogens. In contrast to the acquired genetic alterations described above that are a feature of sporadic ovarian cancers, 5-10% of ovarian cancers probably arise due to inherited genetic defects. Recently, the BRCA1 tumor suppressor gene has heen identified and shown to be responsible for most cases of hereditary ovarian cancer. Further studies are needed to augment our understanding of the molecular pathogenesis of ovarian cancer. PMID- 9337070 TI - Interaction of the DNA topoisomerase II catalytic inhibitor meso-2,3-bis(3,5 dioxopiperazine-1-yl)butane (ICRF-193), a bisdioxopiperazine derivative, with the conserved region(s) of eukaryotic but not prokaryotic enzyme. AB - ICRF-193 [meso-2,3-bis(3,5-dioxopiperazine-1-yl)butane], a bisdioxopiperazine compound, has been shown to be a catalytic inhibitor of DNA topoisomerase II by stabilizing the enzyme in the form of a closed "protein clamp," an intermediate form in the catalytic cycle (Roca et al., Proc Natl Acad Sci USA 91: 1781-1785, 1994). In view of its usefulness as a probe in the functional analysis of the enzyme, we tried further to define the domain(s) of the enzyme interacting with the drug by examining its inhibitory activity on type II topoisomerases from various species of eukaryotes and prokaryotes. ICRF-193 inhibited the enzyme from yeast, fly, frog, plant, and mammals at IC50 values in the range of 1-13 microM. Experiments using fission yeast truncated mutant type II enzyme lacking both amino-terminal 74 amino acids and carboxy-terminal 265 amino acids revealed that ICRF-193 interacts with the 125 kDa "core" polypeptide of the enzyme. In contrast, prokaryotic type II enzymes, Escherichia coli DNA gyrase, topo IV, and phage T4 topo, were not affected by the drug. From these results, the domain(s) common to eukaryotic but not to prokaryotic type II enzymes interacting with ICRF 193 was speculated. PMID- 9337071 TI - Functionally nonequivalent interactions of guanosine 5'-triphosphate, inosine 5' triphosphate, and xanthosine 5'-triphosphate with the retinal G-protein, transducin, and with Gi-proteins in HL-60 leukemia cell membranes. AB - G-proteins mediate signal transfer from receptors to effector systems. In their guanosine 5'-triphosphate (GTP)-bound form, G-protein alpha-subunits activate effector systems. Termination of G-protein activation is achieved by the high affinity GTPase [E.C. 3.6.1.-] of their alpha-subunits. Like GTP, inosine 5' triphosphate (ITP) and xanthosine 5'-triphosphate (XTP) can support effector system activation. We studied the interactions of GTP, ITP, and XTP with the retinal G-protein, transducin (TD), and with G-proteins in HL-60 leukemia cell membranes. TD hydrolyzed nucleoside 5'-triphosphates (NTPs) in the order of efficacy GTP > ITP > XTP. NTPs eluted TD from rod outer segment disk membranes in the same order of efficacy. ITP and XTP competitively inhibited TD-catalyzed GTP hydrolysis. In HL-60 membranes, the chemoattractants N-formyl-L-methionyl-L leucyl-L-phenylalanine (fMLP) and leukotriene B4 (LTB4) effectively activated GTP and ITP hydrolysis by Gi-proteins. fMLP and LTB4 were at least 10-fold more potent activators of ITPase than of GTPase. Complement C5a effectively activated the GTPase of Gi-proteins but was only a weak stimulator of ITPase. The potency of C5a to activate GTP and ITP hydrolysis was similar. The fMLP-stimulated GTPase had a lower Km value than the fMLP-stimulated ITPase, whereas the opposite was true for the Vmax values. fMLP, C5a, and LTB4 did not stimulate XTP hydrolysis. Collectively, our data show that GTP, ITP, and XTP bind to G-proteins with different affinities, that G-proteins hydrolyze NTPs with different efficacies, and that chemoattractants stimulate GTP and ITP hydrolysis by Gi-proteins in a receptor-specific manner. On the basis of our results and the data in the literature, we put forward the hypothesis that GTP, ITP, and XTP act as differential signal amplifiers and signal sorters at the G-protein level. PMID- 9337073 TI - Thiamine deficiency in cardiac cells in culture. AB - Rat heart cells in culture were found to be a unique model for studying biochemical and pharmacological aspects of thiamine deficiency. When thiamine was excluded from the growth medium, the following effects were observed: (1) Morphological examination did not show any difference between control and thiamine-deprived cells during the first 10 days. However, after 10-11 days spontaneous contractions ceased, accompanied by initiation of cell degeneration; (2) Intensive degeneration and cell death were observed after 14-16 days. (3) Thiamine pyrophosphate (TPP) concentration in thiamine-deprived cells was decreased gradually, with an elimination half-life of 4-5 days. (4) [3H]deoxyglucose uptake by the cells was increased, even after 1 day of thiamine deprivation. (5) ATP level decreased after 8 days and reached 50% of control cells after 10 days. (6) In thiamine-deprived cells, thiamine addition caused a 60% rise in contraction amplitude but contraction rate was not altered significantly. (7) All these effects were reversible if thiamine was supplied before the initiation of the degeneration processes. PMID- 9337072 TI - Potentiation of ara-C-induced apoptosis by the protein kinase C activator bryostatin 1 in human leukemia cells (HL-60) involves a process dependent upon c Myc. AB - The role of the nuclear phosphoprotein c-Myc has been examined with respect to the regulation of 1-beta-D-arabinofuranosylcytosine (ara-C)-induced apoptosis in human leukemia cells exposed to bryostatin 1 and other pharmacologic protein kinase C (PKC) activators. Pretreatment of HL-60 cells for 24 hr with 10 nM bryostatin 1 significantly potentiated the ability of ara-C (10 microM; 6 hr) to induce apoptosis without reducing the expression of c-Myc protein. In contrast, equivalent exposure to the stage 2 tumor-promoting PKC activator mezerein (10 nM) in conjunction with ara-C reduced c-Myc levels by 87% and failed to potentiate apoptosis. Co-administration of bryostatin 1 with mezerein before ara-C prevented down-regulation of c-Myc and augmented cell death, whereas co-treatment with the calcium ionophore A23187 (250 nM) and bryostatin 1 reduced c-Myc levels by 80% and abrogated the increase in ara-C-induced apoptosis. When cells were exposed for 24 hr to a c-myc antisense oligonucleotide (AS-ODN;10 microM) but not to a scrambled sequence ODN (SS-ODN) prior to ara-C, c-Myc expression was reduced by 81%, and apoptosis and cell viability were unperturbed. However, AS-ODN (but not SS-ODN) reduced c-Myc protein in cells pre-exposed to bryostatin 1 by 74% and abrogated potentiation of ara-C-induced apoptosis. The actions of c-myc AS-ODN did not stem from proximal G1 arrest/differentiation or biochemical events, since they were not associated with a reduction in the S-phase cell fraction, p21(WAF1/CIP1) induction, pRb hypophosphorylation, or alterations in ara-C metabolism. Together, these findings indicate that HL-60 cell apoptosis proceeds by both c-Myc-dependent and -independent pathways, and that only the former are involved in the potentiation of ara-C-mediated cell death by bryostatin 1. PMID- 9337074 TI - Regulation of A1 adenosine receptors by amiodarone and electrical stimulation in rat myocardial cells in vitro. AB - The effects of conditions that either increase or decrease heart rate on the pharmacological properties of adenosine receptors in cultured rat myocytes were examined. Levels of A1 adenosine receptors, following prolonged treatment with electrical stimulation (ES) or the antiarrhythmic drug amiodarone, were determined using radioligand binding with the specific A1 receptor antagonist [3H]1,3-dipropyl-8-cyclopentylxanthine (CPX). The effects of lowering temperature were also explored. Exposure to amiodarone for 4 days reduced the density of A1 receptors by 19% (from 24.7 +/- 0.4 to 20.09 +/- 0.3 fmol/dish) and inhibited the rate of contraction by 60% (from 188 +/- 16 to 76 +/- 30 beats/min), without changing the receptor affinity, protein content, creatine kinase (CK) activity or cell number. Electrical stimulation at 25 degrees C elevated the density of A1 adenosine receptors by 185% (from 4.1 +/- 0.4 to 11.69 +/- 2.1 fmol/dish). Four days of reduced temperature (from 37 degrees C to either 30 or 25 degrees C) lowered the density of A1 adenosine receptors by 69 or 86%, respectively (from 24.1 +/- 1.2 to 7.4 +/- 0.4 or 3.4 +/- 0.3 fmol/dish), with no significant change in the receptor affinity, activity of CK, or lactate dehydrogenase (LDH), protein content or cell number. The observed up- and down-regulation of A1 adenosine receptors in primary myocyte cultures in response to conditions that exogenously alter the rate of contraction, is indicative of the role of adenosine receptors in adaptation of heart cells to stress. PMID- 9337075 TI - Lamivudine (3TC) phosphorylation and drug interactions in vitro. AB - Lamivudine (2'-deoxy-3'-thiacytidine; 3TC) is a dideoxynucleoside analogue that inhibits the replication of human immunodeficiency virus (HIV). We are currently investigating the intracellular metabolism of 3TC to its active triphosphate (3TCTP) in peripheral blood mononuclear cells (PBMC) and a monocytic cell line (U937). Optimal phosphorylation of 3TC was achieved after incubation for 24 hr, with 3TC diphosphate (3TCDP) the predominant metabolite formed, in both cell types investigated. Further studies in PBMCs followed preincubation with the mitogen phytohaemagglutinin (PHA) for 72 hr. This enabled greater detection of phosphates, compared to resting cells. A 3TC concentration of 1 microM was chosen for future interaction studies, allowing good detection of 3TC and phosphates on radiochromatograms whilst being similar to the plasma level found in clinical studies (i.e. 3 microM). With a shift in treatment to combination therapy, it is essential that potential interactions between nucleoside analogues are investigated at the phosphorylation level, as this could affect antiviral activity. Both deoxycytidine (dC) and 2',3'-dideoxycytidine (ddC) significantly inhibited 3TC phosphorylation (e.g. at dC 100 microM, no 3TCTP was detected in PBMCs; P < 0.001, whereas 66% of control 3TCTP production was observed in U937 cells; P < 0.01). Zidovudine (ZDV) caused a small but significant reduction of 3TC phosphate production in both PBMCs and U937 cells. However, this may be due to toxicity or an effect on endogenous dCTP pools. Neither 2',3'-dideoxyinosine (ddI) or 2',3'-didehydro-2',3'-dideoxythymidine (d4T) significantly inhibited 3TC phosphorylation. These results suggest it would be better to coadminister two nucleoside analogues with different activation pathways. PMID- 9337076 TI - Studies on adaptation to adriamycin in cells pretreated with hydrogen peroxide. AB - Various investigations have reported the occurrence in bacterial and mammalian cells of an adaptive response to the toxic effects of oxidants or agents that cause oxidation via redox reactions. In our previous study, it was shown that several cell lines pretreated with a low dose of hydrogen peroxide (H2O2) exhibited an adaptive response to subsequent high doses of adriamycin (ADR), whereas other cell lines did not. Based on the observation that the cell lines utilized differed in their sensitivity towards adriamycin, we undertook the present investigation with the goal of evaluating possible relationships between the levels of antioxidant enzymes and sensitivity towards adriamycin. Another aim was to determine relationships between the inducibility of these enzymes and the occurrence of adaptation. We utilized African Green monkey kidney (V3), human embryo (CLV98), human melanoma (ME18), and Chinese hamster ovary (CHO) cell lines and experimentally developed adriamycin-resistant human melanoma (ME18/RN) and Chinese hamster ovary (CHO/RN) cell sublines. Cytotoxicity was measured by MTT (3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and trypan blue exclusion. The levels of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were determined in the same kind of experiment as that revealing the occurrence of adaptation. The rank order established for catalase activities was similar to that for sensitivity towards adriamycin. Aberrant increases in the tested enzymes were demonstrated in experimental groups of all kinds of cells. We conclude that in our cell systems catalase is a major determinant of adriamycin resistance. Whether the occurrence of the adaptive response under study is dependent on the contribution of catalase, itself dependent on the degree of resistance to the drug, is discussed. PMID- 9337077 TI - In vitro metabolism of dexamethasone (DEX) in human liver and kidney: the involvement of CYP3A4 and CYP17 (17,20 LYASE) and molecular modelling studies. AB - Dexamethasone (DEX) has previously been shown to be extensively metabolised to 6 hydroxylated and side-chain cleaved metabolites in human liver in vitro. CYP3A4 is responsible for 6alpha- and 6beta-hydroxylation of DEX and CYP17 is thought to mediate side-chain cleavage to generate 9alphafluoro-androsta-1,4-diene-11beta hydroxy-16alpha-methyl-3,17-dione (9alphaF-A). Although 9alphaF-A has not previously been isolated as a metabolite in its unhydroxylated form in human liver incubations, it is formed as an intermediate metabolite, which is subsequently rapidly hydroxylated to OH-9alphaF-A. A main part of this study has been to conclusively show that DEX undergoes extensive side-chain cleavage to form 9alphaF-A in human kidney fractions, which is in contrast to profiles obtained for DEX metabolism in parallel human liver microsomal incubations where 6-hydroxylation is the predominant pathway. Furthermore, molecular models of CYP3A4 and CYP17 (17,20 lyase) have been used to model the enzyme fits of DEX. From these modelling studies it has been shown that DEX complements both putative enzyme active sites in orientations likely to lead to the formation of the metabolites identified in vitro. We have also been able to rationalise the preferential formation of the 6betaOH-DEX isomer. PMID- 9337078 TI - Reduction of bone loss by denbufylline, an inhibitor of phosphodiesterase 4. AB - The effects of denbufylline, a xanthine derivative with selective inhibitory activity on the phosphodiesterase (PDE) 4 isoenzyme, on bone loss in Walker 256/S bearing rats and on mineralized nodule formation and osteoclastlike cell formation in bone marrow culture systems were examined. Serial oral administrations of denbufylline inhibited the decrease in the bone mineral density of femurs from Walker 256/S-bearing rats, without influence on the healthy rats. Denbufylline restored the bone mass and the number of osteoclasts and osteoblasts per trabecular surface in the femur metaphysis. Among PDE inhibitors, only PDE4-selective inhibitors increased the number of mineralized nodules and decreased the number of osteoclastlike cells in the in vitro bone marrow culture systems, and dibutyryl cyclic AMP mimicked these effects in the in vitro systems. These results suggest that the PDE4 isoenzyme may play an important role in bone turnover through cyclic AMP and that its inhibitors are candidates for therapeutic drugs for the bone loss diseases. PMID- 9337079 TI - Renal excretory responses to saline load in the taurine-depleted and the taurine supplemented rat. AB - Taurine is found in high concentrations in mammalian cells. Despite recognition of its role as an organic osmolyte in the kidney, information regarding its effects on renal fluid and electrolyte excretion is sparse. Therefore, the objective of the first series of experiments was to determine the effects of taurine depletion on renal excretory responses to a saline load. To induce taurine depletion, male Wistar-Kyoto (WKY) rats were treated with tap water containing 3% beta-alanine for 3 weeks. Taurine depletion reduced the initial rates of fluid and sodium excretion after an intravenous saline load. This effect was attributed to taurine depletion since maintenance of the taurine-depleted rats on tap water for 2 days to remove the effects of beta-alanine yielded the same pattern as the taurine-depleted rats exposed to beta-alanine at the time of the experiment. Nonetheless, rats exposed to short-term beta-alanine treatment, which has no influence on kidney taurine content, demonstrated a larger (approximately 25%) natriuretic but not diuretic response to the isotonic saline load than either the control or taurine-depleted rats. These data suggest that beta-alanine-induced inhibition of tubular reabsorption of taurine may result in subsequent excretion of taurine with attendant natriuresis early in the course of beta-alanine treatment. We also tested the hypothesis that taurine potentiates the renal excretory responses to an isotonic saline load in WKY rats. Inclusion of taurine in the infusate significantly increased natriuresis and diuresis after a saline load. This effect was greater in animals fed a basal than a high NaCl diet. Our data support a role for taurine as a natriuretic and diuretic agent. PMID- 9337080 TI - The role of vitamin D derivatives and retinoids in the differentiation of human leukaemia cells. AB - The capabilities of 1alpha, 25-dihydroxyvitamin D3 (1,25(OH)2D3), and two novel vitamin D analogues, EB1089 and KH1060, to induce the differentiation of two established leukaemia cell lines, U937 and HL-60, were assessed alone or in combination with the retinoid compounds, 9-cis retinoic acid (9-cis RA) and all trans retinoic acid (ATRA). The vitamin D derivatives acted to increase the differentiation of U937 and HL-60 cell cultures in a dose-dependent manner, as determined by nitroblue tetrazolium (NBT) reduction, with EB1089 and KH1060 being more effective than the native hormone. As an additional index of leukaemic cell differentiation, induction of expression of the phenotypic cell surface antigen, CD14, and the beta2-integrins, CD11b and CD18 by the vitamin D and retinoid compounds were monitored using fluorescence activated cell sorting (FACS) analyses. Following 96-hr treatment of U937 and HL-60 cells with 5 x 10(-10) M of the vitamin D derivatives, a striking increase in CD14 antigen expression was apparent, indicating the promotion by these compounds of a monocyte/macrophage lineage of cells. CD11b and CD18 antigen expression were also raised above control levels. In contrast, both retinoid compounds used at the higher concentration of 1 x 10(-8) M were not effective inducers of CD14 antigen expression. However, CD11b and CD18 were both readily increased in U937 and HL-60 cell cultures. Treatment of U937 cell cultures with the vitamin D compounds and the retinoids resulted in cooperative effects on induction of differentiation, with correlation by both NBT reduction and FACS analyses of CD14 antigen expression. The presence of 9-cis RA or ATRA appeared to contribute to the further increase of CD14 in these cells. HL-60 cell cotreatment with these compounds also displayed enhanced cooperative effects in phagocytic function by NBT reduction. However, analysis of CD14 revealed a dramatic diminution in HL-60 cells treated with the combinations of the vitamin D derivatives and the retinoids. Assessment of HL-60 cell morphology treated with these combinations demonstrated the presence of a mixed population of monocytes and granulocytes. CD11b and CD18 antigen expression was also enhanced in both cell lines with cotreatment. The ability of EB1089 and KH1060 to induce leukaemic cell differentiation may provide an additional option for therapeutic use alone or together with other differentiation agents such as 9-cis RA or ATRA. PMID- 9337081 TI - Measurement of deoxyuridine triphosphate and thymidine triphosphate in the extracts of thymidylate synthase-inhibited cells using a modified DNA polymerase assay. AB - New inhibitors of the enzyme thymidylate synthase (TS) are now reaching clinical application. Alteration of the dUTP: dTTP ratio may be critical to TS inhibition induced tumor cell death. The DNA polymerase assay with modification was used to rapidly and sensitively measure dUTP, dTTP, and dUTP:dTTP ratios in cell extracts of HT29 human colon carcinoma cells treated with the specific TS inhibitor ZD1694 [N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2 thenoyl)-L-glutamic acid]. These results revealed an increase in the dUTP:dTTP ratio at 2 hr after a 2-hr exposure to ZD1694 at concentrations of 0.05 to 0.2 microM with significant normalization at 16 hr after a 2-hr exposure despite evidence of continued TS inhibition. This assay is highly sensitive and reproducible for levels of dUTP and is less labor intensive than traditional assays. PMID- 9337082 TI - Monoclonal antibodies for the detection of desialylation of erythrocyte membranes during haemolytic disease and haemolytic uraemic syndrome caused by the in vivo action of microbial neuraminidase. AB - Especially in childhood, the in vivo action of microbial neuraminidase may cause haemolytic anaemia or life-threatening haemolytic uraemic syndrome. The exposure of the Thomsen-Friedenreich (T) crypto-antigen and T-antigen polyagglutinability of erythrocytes has been described as the first sign of toxic cleavage of N acetylneuraminic acid (Neu5Ac) from sialoglycoproteins of cell membranes. This phenomenon may, however, be too unspecific to initiate treatment for toxin elimination. The present study investigated the diagnostic effectiveness of a panel of three monoclonal antibodies (mcabs) for the estimation of the clinical significance of neuraminidase action in vivo. Depending on the amount of Neu5Ac released, the mcabs I-C4, II-Q9 and III-Y12 recognized different epitopes on erythrocyte asialoglycophorin. In 1345 patients, the mcab II-09 detected cleavage of Neu5Ac in 32 children who had T-antigen polyagglutinability and mild to moderate haemolytic anaemia. However, only 10 patients, whose erythrocytes were agglutinated by the mcabs III-Y12 or I-C4, developed severe haemolysis, thrombocytopenia, and finally the life-threatening haemolytic uraemic syndrome (p<0.0002). In conclusion, these mcabs provided an early marker of the in vivo action of neuraminidase. Two different degrees of erythrocyte desialylation, as defined by these mcabs, are suggested to reflect the severity of toxin-associated disease. PMID- 9337083 TI - Stimulatory effect of gangliosides on phagocytosis, phagosome-lysosome fusion, and intracellular signal transduction system by human polymorphonuclear leukocytes. AB - Gangliosides are known to be differentiation-inducing molecules in mammalian stem cells. We studied the interaction between the molecular structure of glycosphingolipids (GSLs) and their promoting mechanisms of the phagocytic processes in human polymorphonuclear leukocytes (PMN). The effect of various gangliosides from mammalian tissues on adhesion, phagocytosis, phagosome-lysosome (P-L) fusion and superoxide anion production was examined by human PMN using heat killed cells of Staphylococcus aureus-coated with GSLs. Gangliosides GM3, GD1a, GD3 and GT1b showed a marked stimulatory effect on the phagocytosis and P-L fusion in a dose-dependent manner, while ganglioside GM1, asialo GM1 and neutral GSLs did not. The relative phagocytic rate of ganglioside GM3-coated S. aureus was the highest among the tested GSLs. Both P-L fusion rate and phagocytosis of S. aureus were elevated significantly when coated with ganglioside GD1a, GD3 or GT1b, and GT1b gave a five times higher rate than that of the non-coated control. These results suggest that the terminal sialic acid moiety is essential for the enhancement of phagocytosis and that the number of sialic acid molecules in the ganglioside is related to the enhancement of the P-L fusion process. On the other hand, the superoxide anion release from PMN was not affected by ganglioside GM2, GM3, GD1a or GT1b. Furthermore, to clarify the trigger or the signal transduction mechanism of phagocytic processes, we examined the effect of protein kinase inhibitors such as H-7, staurosporine (protein kinase C inhibitor), H-89 (protein kinase A inhibitor), genistein (tyrosine kinase inhibitor), ML-7 (myosin light chain kinase inhibitor), and KN-62 (Ca2+/calmodulin-dependent protein kinase II inhibitor) on ganglioside-induced phagocytosis. H-7, staurosporine and KN-62 inhibited ganglioside-induced phagocytosis in the range of concentration without cell damage, while H-89, genistein and ML-7 did not. Moreover, H-7 and KN-62 inhibited ganglioside-induced P-L fusion. These results suggest that protein kinase C and Ca2+/calmodulin-dependent protein kinase II may be involved in the induction of phagocytosis and P-L fusion stimulated by gangliosides. PMID- 9337084 TI - Expression and sulfogalactolipid binding specificity of the recombinant testis specific cognate heat shock protein 70. AB - Immunofluorescent studies with anti-2A antisera, raised specifically against a synthetic C-terminal peptide of native murine P70, the testes-specific cognate heat shock protein 70, demonstrated that the rat homologue of P70 is expressed on the surface of testicular cells. The murine hsp 70.2 gene, encoding P70, was cloned and expressed in Escherichia coli. The recombinant P70 (rP70) protein with a 6Xhistidine affinity tag at its amino terminus was purified from E. coli via nickel affinity column chromatography. Monoclonal anti-hsp70 antisera and anti-2A antisera cross-reacted with purified rP70. Binding of rP70 was specific for sulfogalactosylceramide (SGC) and sulfogalactosyglycerolipid (SGG). Binding was not inhibited by the sugar, galactose 3'sulfate, nor was binding observed to desulfated derivatives of SGC and SGG, to other negatively charged lipids or other sulfated lipids. Furthermore, rP70 bound to an SGC-column and was eluted only at high salt in combination with high pH. These results show rP70 to possess a specific sulfatide binding site. Since the biochemical properties and immunoreactivity of rP70 are indistinguishable from native P70 and SLIP1 (testicular sulfoglycolipid immobilized protein 1) rP70 can be employed to examine the role of hsp70-mediated sulfatide binding in fertilization. PMID- 9337085 TI - Anti-inflammatory activity of superoxide dismutase conjugated with sodium hyaluronate. AB - Superoxide dismutase (SOD) from bovine erythrocytes was conjugated with sodium hyaluronate (HA) with a mean molecular weight of 10(6) to have greater anti inflammatory activity in vivo. Amino groups of SOD were coupled with carboxyl groups in the hyaluronate molecule using 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide. The HA-SOD conjugate was composed of 1.5 mol of SOD molecule per 1 mol of hyaluronate on the average, and retained 70% of the activity of unmodified SOD. The conjugate was essentially non-immunogenic in mice, and exhibited much higher anti-inflammatory activities than HA or SOD in models of inflammatory diseases such as ischemic oedema of the foot-pad in mice, carrageenin-induced pleurisy and adjuvant arthritis in rats. PMID- 9337086 TI - Beta-galactosidase-deficient mouse as an animal model for GM1-gangliosidosis. AB - GM1-gangliosidosis is a progressive neurological disease in humans caused by deficiency of lysosomal acid beta-galactosidase, which hydrolyses the terminal beta-galactosidic residue from ganglioside GM1 and other glycoconjugates. In this study, we generated a mouse model for GM1-gangliosidosis by gene targeting in embryonic stem cells. The mouse homozygous for the disrupted beta-galactosidase gene showed beta-galactosidase deficiency, presented with progressive spastic diplegia, and died of emaciation at 7-10 months of age. Pathologically, PAS positive intracytoplasmic storage was observed in neuronal cells of various areas in the brain. Biochemical analysis revealed a marked accumulation of ganglioside GM1 and asialo GM1 in brain tissue. This animal model will be useful for pathogenetic analysis and therapeutic trial of human GM1-gangliosidosis. PMID- 9337087 TI - Assessment of glycosaminoglycan-protein linkage tetrasaccharides as acceptors for GalNAc- and GlcNAc-transferases from mouse mastocytoma. AB - Two glycosaminoglycan-protein linkage tetrasaccharide-serine compounds, GlcAbeta1 3Galbeta1-3Galbeta1-4Xylbeta1-O-Ser and GlcAbeta1-3Gal(4-O-sulfate)beta1 3Galbeta1-4Xylbeta1-O -Ser, were tested as hexosamine acceptors, using UDP [3H]GlcNAc and UDP-[3H]GalNAc as sugar donors, and solubilized mouse mastocytoma microsomes as enzyme source. The nonsulfated Ser-tetrasaccharide was found to function as an acceptor for a GalNAc residue, whereas the Ser-tetrasaccharide containing a sulfated galactose unit was inactive. Characterization of the radio labelled product by digestion with alpha-N-acetylgalactosaminidase and beta-N acetylhexosaminidase revealed that the [3H]GalNAc unit was alpha-linked, as in the product previously synthesized using serum enzymes, and not beta-linked as found in the chondroitin sulfate polymer. Heparan sulfate/heparin biosynthesis could not be primed by either of the two linkage Ser-tetrasaccharides, since no transfer of [3H]GlcNAc from UDP-[3H]GlcNAc could be detected. By contrast, transfer of a [3H]GlcNAc unit to a [GlcAbeta1-4GlcNAcalpha1-4]2-GlcAbeta1-4-aMan hexasaccharide acceptor used to assay the GlcNAc transferase involved in chain elongation, was readily detected. These results are in agreement with the recent proposal that two different N-acetylglucosaminyl transferases catalyse the biosynthesis of heparan sulfate. Although the mastocytoma system contains both the heparan sulfate/heparin and chondroitin sulfate biosynthetic enzymes the Ser tetrasaccharides do not seem to fulfil the requirements to serve as acceptors for the first HexNAc transfer reactions involved in the formation of these polysaccharides. PMID- 9337122 TI - Progressive changes in cortical metabolites at three stages of infantile hydrocephalus studied by in vitro NMR spectroscopy. AB - Infantile hydrocephalus is most often caused by an obstruction in the cerebrospinal fluid flow pathway and results in ventricular dilatation and chronic trauma to the surrounding brain. Surgical treatment alleviates the condition but does not cure or prevent neurological deficits. The H-Tx rat has severe hydrocephalus due to a spontaneous aqueduct obstruction in late gestation. In order to determine how hydrocephalus affects brain metabolism in tissue adjacent to the expanded ventricles, cortical extracts have been made from groups of hydrocephalic and control littermates with early, intermediate, and advanced hydrocephalus at 4, 11, and 21 days after birth. Extracts were analyzed with 1H and 31P NMR spectroscopy and metabolite peaks were quantified using an external standard. Metabolite concentrations were calculated relative to tissue wet weight and subsequently expressed relative to tissue dry weight, using values for water content obtained from additional groups of rats. In early hydrocephalus there was a significant decrease in the phosphomonoester phosphorylcholine, and there were small, nonsignificant changes in other compounds. By 11 days, in addition to phosphomonoesters, there were significant decreases in ATP, phosphocreatine, and in inorganic phosphate, but with no change in lactate. By 21 days there were also substantial decreases in cholines, inositol, creatine, glutamate, glutamine, aspartate, N-acetylaspartate, alanine, and taurine. It is concluded that the sequence of pathological events starts with changes in membrane lipids. This is followed by reductions in energy metabolite which leads to cell swelling with loss of intracellular osmolytes and neurotransmitters. These changes are discussed in relation to hydrocephalus pathophysiology and to prevention and reversibility with shunt treatment. PMID- 9337123 TI - Loss of axonal microtubules and neurofilaments after stretch-injury to guinea pig optic nerve fibers. AB - Axonal swellings, characterized by focal accumulations of membranous organelles at presumed sites of interrupted axonal transport, occur in diffuse axonal injury (DAI) in human, blunt head injury and in animal models of nondisruptive axonal injury. Membranous organelles are transported by fast axonal transport in association with microtubules. Although loss of microtubules has been documented at levels of injury severe enough to result in permeabilization of the axolemma to tracers such as horseradish peroxidase, there has been no detailed analysis of responses by microtubules in less severe or milder forms of nondisruptive axonal injury. To test the hypothesis that in less severe forms of axonal injury there is a rapid response by axonal microtubules that might provide an explanation for loss of fast axonal transport, we have carried out a morphometric analysis of microtubules in CNS axons after stretch-injury. There is loss of microtubules at nodes of Ranvier with nodal blebs within 15 min of injury, and in internodal axonal swellings between 2 and 4 h. There is a return to control values at nodes of Ranvier by 4 h, and at the internode by 24 h. There is no loss of microtubules at paranodes, although there is a reduction in their density in the first 2 h after injury. The greatest loss of microtubules occurs at sites of axolemma infolding. Hypothetical mechanisms that might lead to this loss resulting in focal disruption of fast axonal transport and the formation of axonal swellings are discussed. PMID- 9337124 TI - Morris water maze deficits in rats following traumatic brain injury: lateral controlled cortical impact. AB - This experiment utilized a laterally placed controlled cortical impact model of traumatic brain injury (TBI) to assess changes on spatial learning and memory in the Morris water maze (MWM). Adult rats were subjected to one of two different levels of cortical injury, mild (1 mm) or moderate (2 mm) deformation, and subsequently tested for their ability to learn (acquisition) or remember (retention) a spatial task, 7 or 14 days after injury. Results revealed an injury dependent deficit for experimental animals compared to sham-operated controls. Not only did the TBI result in longer escape latencies, but also significant deficits in search time and relative target visits. Although the moderately injured animals demonstrated significant histopathology in the cortex and hippocampus, mildly injured subjects demonstrated no obvious tissue destruction, but did manifest significant behavioral change. These results demonstrate that a laterally placed controlled cortical impact is capable of producing significant cognitive deficits on both acquisition and retention paradigms utilizing the MWM. PMID- 9337125 TI - Transiently increased basilar artery flow velocity following severe head injury: a time course transcranial Doppler study. AB - BACKGROUND AND PURPOSE: Transcranial Doppler ultrasonography has been used to study changes in cerebral hemodynamics following head injury. However, most studies evaluated the anterior circulation and little information exists on transcranial Doppler of the vertebrobasilar arteries after head injury. METHODS: Thirty-two patients with a Glasgow Coma Scale (GCS) score between 4-8 and 11 patients with a GCS score between 9-14 were studied using transcranial Doppler ultrasonography for the first 10 days after injury. Daily variations in the mean blood flow velocities of all major cerebral arteries were recorded. RESULTS: In patients with GCS score between 4-8, the mean blood flow velocities in the middle cerebral and basilar arteries gradually increased beginning on day 2 postinjury and peaked on the 4th-5th day after injury. Those changes were more prominent, and appeared earlier, in the basilar artery. The ratio between the mean flow velocities of the middle cerebral artery and the basilar artery during the first 4 days was significantly lower than in normal controls, indicating a particular increase of flow velocity in the basilar artery. Nineteen out of 32 patients (60%) with severe head injury showed mean blood flow velocity increased over 75 cm/sec in the basilar artery. Mean blood flow velocity >90 cm/sec in the basilar artery, compatible with vasospasm, was observed in 12 of 32 patients (37%). Spasm in the middle cerebral artery was observed in 12 (37%) of patients; 10 of them also had evidence of basilar artery spasm. On the whole, 14 of 32 (43%) patients had evidence of spasm either in the middle cerebral or basilar arteries or in both. In 5 of 11 patients (50%) with moderate head injury (GCS score 9-14), blood flow velocity in the basilar artery greater than 75 cm/sec was observed, but in only two of them it reached the values over 90 cm/sec. Vasospasm in the middle cerebral artery was noted in one patient. CONCLUSIONS: A significant number of patients develop increased flow velocities compatible with vasospasm in the basilar artery after severe head injury. This phenomenon may represent an additional factor that contributes to the poor outcome of severely head-injured patients. PMID- 9337126 TI - Indomethacin in the management of elevated intracranial pressure: a review. AB - Elevated intracranial pressure occurs frequently in patients with severe head injury. A number of studies in recent years suggest that indomethacin may be useful in the management of elevated intracranial pressure. Indomethacin acts primarily by reducing cerebral blood flow and decreasing cerebral edema following head injury. This review summarizes the basic and clinical studies of the effects of indomethacin on cerebral blood flow, brain edema, and intracranial pressure. The pharmacology of indomethacin, and issues for future investigation in the use of indomethacin in severe head injury, are discussed. PMID- 9337127 TI - New in vitro model of traumatic neuronal injury: evaluation of secondary injury and glutamate receptor-mediated neurotoxicity. AB - The multiplicity and complexity of secondary injury processes following brain trauma in vivo make it difficult to elucidate the roles of specific injury mechanisms. As with other areas of CNS injury, such as ischemia, this has led to the development of in vitro models. Here we describe a new trauma model, in which standardized trauma is delivered to neuronal/glial cultures using a special mechanical device that produces concentric circular cuts in the cell layer. Changes in the number of circles (from 1 to 6) allows variation of injury severity. Comparison studies of cell death induced by such trauma in glial and neuronal/glial cultures demonstrated that glial cells are relatively resistant to this injury, and that the cell death after trauma to neuronal/glial cultures reflects primarily neuronal death. Consistent with other in vivo and in vitro studies, glutamate receptor antagonists MK 801 and MCPG were neuroprotective. Thus, this model appears useful for studying glutamatergic mechanisms involved in secondary injury, and may prove useful for evaluating certain pharmacological strategies for CNS trauma. PMID- 9337128 TI - NMDA-receptor antagonist protects neurons from secondary degeneration after partial optic nerve crush. AB - Damage resulting from a partial acute lesion of white matter in the central nervous system (CNS) gradually spreads also to neurons that escaped the primary injury, resulting in their degeneration. Such spreading has been referred to as secondary degeneration. In order to demonstrate that this degeneration is indeed secondary to that caused by the acute insult, as well as to investigate the mechanism underlying the spread of damage and ways in which to protect neurons from such damage, we have proposed the use of partial lesion of the rodent optic nerve as a model. In this model we examined whether an antagonist of a receptor mediated channel, shown to be beneficial in gray matter lesions, can protect neurons from undergoing secondary degeneration following white matter lesion. A well-calibrated partial crush lesion inflicted on the optic nerve of adult rats was immediately followed by a single intraperitoneal injection of the N-methyl-D aspartate receptor antagonist, MK-801 (1 mg/kg). Protection of neurons from secondary degeneration was assessed by retrograde labeling and by measurement of the visual evoked potential (VEP) response to light. Two weeks after the injury, the mean number of neurons that were still intact was about threefold higher in the MK-801-treated group than in the saline-treated control group, indicating a treatment-induced protection of neurons that had escaped primary injury. A positive VEP response to light was obtained in 90% of the MK-801 treated animals and in only 50% of injured controls. The questions regarding whether the secondary degeneration of initially spared neurons starts in their cell bodies or in their axons, and consequently the identity of the primary site of their protection by MK-801, are discussed in relation to the absence of N-methyl-D aspartate receptors on nerve fibers. The present findings may have implications for both acute and chronic injuries of the CNS. PMID- 9337129 TI - Expression of the Sox11 gene in mouse embryos suggests roles in neuronal maturation and epithelio-mesenchymal induction. AB - Sry, the mammalian Y-linked testis determining gene, is a member of a family of genes known as Sox genes, which encode transcription factors related by a DNA binding motif termed the HMG box. Sox genes are known to have diverse roles in vertebrate differentiation and development. We report here the cloning and characterisation of one of these genes, Sox11, in mice. In addition to an N terminal HMG box domain, the deduced SOX11 protein contains a number of highly conserved C-terminal motifs, which may function in transcriptional regulation. Expression of Sox11 in mouse embryos was prominent in the periventricular cells of the central nervous system, suggesting a role in neuronal maturation. Expression was also observed in a wide range of tissues involved in epithelial mesenchymal interactions, suggesting an additional role in tissue modelling during development. PMID- 9337130 TI - Expression of the metastasis-associated mts1 gene during mouse development. AB - The mts1 gene, a member of the S100 family, is specifically expressed in different metastatic tumor cell lines. After transfection in some nonmetastatic cell lines Mtsl can induce a metastatic phenotype. Mts1 protein can interact with non-muscle myosin, indicating that Mts1 plays a role in cell motility. In order to understand the function of this gene, we studied the expression of the mts1 mRNA and protein in vivo during mouse development. Both mRNA and protein were present in high concentrations from 12.5 to 18.5 days post coitum (dpc) in a variety of developing embryonic tissue of mesodermal origin. We found by double immunostaining with a macrophage-specific antibody that Mts1 protein was highly expressed in fetal macrophages throughout the embryonic mesenchyme and in macrophages colonizing developing lymphatic and non-lymphatic organs. Moreover, we found mts1 expression during differentiation and morphogenesis of mesenchymal tissues such as the mesenchyme surrounding the tips of digits, the mesenchyme underlying the epithelium of the bladder, and the mesenchyme between the primordia of the nasal capsule and the skin as well as in the developing dermal papilla of hair and tooth follicle. In developing bone, Mts1 was expressed in invasive mesenchymal cells and in osteoclasts. The results presented here suggest that Mtsl plays an important role in mouse development during differentiation and function of macrophages and might be involved in different processes associated with mesenchymal morphogenesis including mesenchymal-epithelial interaction, tissue remodeling, and invasion. PMID- 9337131 TI - Contribution of the primitive epicardium to the subepicardial mesenchyme in hamster and chick embryos. AB - A study about the hypothetical contribution of the epicardial cells to the subepicardial mesenchyme was carried out in Syrian hamster embryos of 9-12 days post coitum (dpc) and chick embryos of 3-5 days of incubation. In the epicardium and subepicardium of these embryos we have immunolocated the proteins cytokeratin (CK), vimentin (VIM), fibronectin (FN), and two antigens related to the transformation of endocardial cells into valvuloseptal mesenchyme, ES/130 and JB3. In the hamster embryos, CK+ subepicardial mesenchymal cells (SEMC) were apparently migrating from the primitive epicardium from 9.5 dpc at the atrioventricular (AV) groove and proximal outflow tract (OFT). The morphological signs of delamination extended by 11 dpc to the epicardium of the interventricular groove and the dorsal part of the ventricle. The relative abundance of the CK+ SEMC decreased in embryos of 12 dpc. VIM colocalized with CK in most SEMC, and in some epicardial mesothelial cells, mainly at the areas of delamination. CK immunoreactivity was also found in some early subepicardial capillaries. Similar observations were made in the chick embryos studied. The immunoreactive patterns obtained at the subepicardium with anti-FN, ES/130, and JB3 antibodies were similar to those reported in the areas of endothelial transformation of the endocardial cushions. We suggest that these observations are compatible with an epithelial-mesenchymal transformation involving the epicardial mesothelium and originating at least a part of the SEMC. PMID- 9337132 TI - Transitory expression of alpha cardiac myosin heavy chain in a subpopulation of secondary generation muscle fibers in the pig. AB - Unlike the random distribution of fiber types seen in skeletal muscles of most mammals, pig muscle exhibits a rosette pattern consisting of islets of slow fibers surrounded by concentric circles of type IIA and IIB fibers. Within each islet of slow fibers, one of the central fibers is a primary myofiber, whereas all others are secondary fibers. The present study demonstrates that a subpopulation of the slow secondary fibers transiently expresses alpha-myosin heavy chain (MHC). Two cDNA libraries were made from longissimus dorsi skeletal muscle of 14-day-old piglet and adult pig atrium; the latter muscle is mainly composed of alpha-MHC. Screening of the libraries with a human anti-alpha-MHC mAb (F8812F8) demonstrated the presence of positive MHC clones in both libraries; the nucleotide sequence of the 3'-untranslated region (3'-UTR) was identical in both libraries. As this MHC 3'-UTR had 75% homology with the human alpha-MHC, it was identified as pig alpha-MHC. Using specific cRNA probes and mAbs against pig alpha-cardiac and beta/slow/type I MHC, we studied the expression of these MHCs in developing pig semitendinosus muscle by combining in situ hybridization and immunocytochemistry on serial sections at 90 days of gestation, and at 1, 6, 35 days and 6 months of age. The results showed that a subpopulation of secondary fibers that directly abut primary fibers, transiently produced alpha-MHC, both at the levels of the protein and its transcript. Subsequently, these fibres expressed beta-MHC. At 1 day, immunocytochemistry showed that 16% of the secondary fibers expressed alpha-MHC, among which 20% did not yet express beta MHC. At 6 days, alpha- and beta-MHCs were mostly present in the same fibers, i.e., 23% of the secondary fibers. Thereafter, the proportion of secondary fibers reacting with alpha-MHC mAb decreased to 10% at 5 weeks and 0% at 6 months, whereas beta-MHC was still accumulating in about 38% of the secondary fibers. During the period studied, the distribution of alpha- and beta-MHC transcripts closely matched that of the corresponding proteins. Expression of alpha-MHC was not detected in primary type I muscle fibers and slow type I secondary fibers at the periphery of the rosettes of slow fibers. This study is the first unequivocal demonstration of a transitory expression of alpha-MHC in a subpopulation of secondary fibers in a limb skeletal muscle during mammalian development. PMID- 9337133 TI - Neurotrophin mRNA expression in the developing tooth suggests multiple roles in innervation and organogenesis. AB - To analyze the roles of neurotrophins during early development of rat teeth, we studied the expression of neurotrophin mRNAs from the initiation of first molar formation to the completion of crown morphogenesis. With RNAase protection assay all neurotrophin mRNAs were detected in embryonic teeth. In situ hybridization analysis revealed developmentally changing, distinct expression patterns for nerve growth factor (NGF) and neurotrophin-3 (NT-3), which were shown not to be regulated by or dependent on peripheral innervation. NGF mRNAs appeared in the mesenchymal target field of the tooth at the time of the trigeminal axon ingrowth (embryonic days 14-15: E14-E15), and they were also present along the pathway taken by growing trigeminal axons. NT-4/5 mRNAs were uniformly expressed in all epithelial cells, but brain-derived neurotrophic factor (BDNF) transcripts were not detected. All neurotrophins induced neurite outgrowth from E13-E16 trigeminal ganglion explants. These results suggest that NGF is involved in the guidance of trigeminal axons to embryonic teeth. In postnatal teeth, expression of NGF mRNAs, but not other neurotrophins, correlated with trigeminal axon ingrowth, proposing that NGF is involved in local sprouting and establishment of the final innervation pattern of the dental papilla and dentin. These results suggest that NGF is required for tooth innervation and that other neurotrophins may also have regulatory roles. In addition, the expression patterns of NGF, NT-3, and NT-4/5 as well as of neurotrophin receptors suggest that the neurotrophin system may also serve non-neuronal functions during tooth development. PMID- 9337134 TI - Developmental expression of perlecan during murine embryogenesis. AB - Perlecan is a modular heparan sulfate proteoglycan that is an intrinsic constituent of all basement membranes and extracellular matrices. Because of its strategic position and unique structure, perlecan has been implicated in modulating the activity of various growth factors required for normal development and tissue homeostasis. To gain insights into the potential function of perlecan in vivo, we examined the spatiotemporal distribution of its mRNA and protein core during murine embryogenesis. We utilized a new affinity-purified antibody that recognizes specifically the protein core of perlecan together with an in situ RT PCR approach to perform a systematic analysis of perlecan expression and deposition during murine ontogeny. Perlecan appeared early (E10.5) in tissues of vasculogenesis including heart, pericardium, and major blood vessels. Its early expression coincided with the development of the cardiovascular system. Subsequently (E11-13), the greatest deposition of perlecan occurred within the developing cartilage, especially the cartilage undergoing endochondral ossification, where it remained elevated throughout all the developmental stages, and up to adulthood. Interestingly, the mRNA levels of perlecan were always higher in all the vascularized tissues, principally within endothelial cells, while chondrocytes displayed relatively low mRNA levels. This suggests a higher biosynthesis and turnover rates in the blood vessels vis-a-vis those of cartilaginous and other mesenchymal tissues. During later stages of development (E13-17.5) perlecan mRNA levels progressively increased and its expression correlated with the onset of tissue differentiation of various parenchymal organs including the developing kidneys, lungs, liver, spleen, and gastrointestinal tract. The central nervous system showed no perlecan expression with the exception of the calvaria and choroid plexus. Collectively, the results indicate that perlecan may play crucial roles not only in vasculogenesis but also in the maturation and maintenance of differentiated tissues, including cartilage. PMID- 9337135 TI - Segmental identity can change independently in the hindbrain and rhombencephalic neural crest. AB - In this study we tested whether the segmental identities of the hindbrain and its derived neural crest are necessarily linked or, instead, if they can be altered independently. Using morphological criteria, we show that the hindbrains of Hoxa 2 mutant mice, in which the second arch skeletal derivatives assume first arch characteristics (Gendron-Maguire et al. [1993] Cell 75:1317-1331; Rijli et al. [1993] Cell 75:1333-1349), retain normal segmental identities. Also, by phenotypic analysis, we show that, with retinoic acid, changes can be induced in the identity of the preotic hindbrain without effects in its derived neural crest. Our data thus indicate that identity changes in the hindbrain and branchial arch neural crest can occur independently. Moreover, if Hoxa-2 is concomitantly induced by retinoic acid in the first branchial arch, the proximal derivatives of this arch are also affected. We propose a model for the patterning of the branchial region, according to which the segmental identity in this area is provided mainly by the branchial arches. PMID- 9337136 TI - Expression of reelin, the gene responsible for the reeler mutation, in embryonic development and adulthood in the mouse. AB - reelin has recently been isolated as a candidate gene, the mutation of which gives rise to the reeler phenotype in mice. In this study, we analyzed the expression of reelin during embryonic development in the mouse and in adult mouse tissues, by in situ hybridization. reelin transcripts were present on embryonic day (E) 8.5 in the somite, foregut, yolk sac, and unclosed neural plate. reelin was expressed in the brain, spinal cord, liver, and kidney throughout embryonic development, and transiently in many developing organs such as the optic cup, blood vessels, precartilage, stomach, pituitary, vibrissae, tooth germ, and in cells along growing nerve fibers. These observations indicate a role for reelin in development of organs in addition to that in neuronal migration. Furthermore, we demonstrated the existence of reelin mRNA and its cellular distribution in the adult brain, spinal cord, liver, kidney, testis, and ovary, suggesting additional roles for reelin in stabilizing the cyto-architecture and in remolding in adult organs. However, we detected no obvious phenotype of the reelin-expressing organs except for the brain in the reeler mouse, indicating the functional redundancy of this gene during the development of these organs. PMID- 9337137 TI - Meis2, a novel mouse Pbx-related homeobox gene induced by retinoic acid during differentiation of P19 embryonal carcinoma cells. AB - We report the cDNA cloning, partial genomic organization, and expression pattern of Stra10, a novel retinoic acid-inducible gene in P19 embryonal carcinoma cells. Four murine cDNA isoforms have been isolated, which are likely to result from alternative splicing. The predicted protein sequences exhibit approximately 85% identity with the Pbx-related Meis1 homeobox gene products, which are involved in myeloid leukemia in BXH-2 mice, and one of the Stra10 isoforms corresponds to the recently published Meis2 sequence (Nakamura et al. [1996] Oncogene 13:2235-2242). The Meis2 homeodomain is identical to that of Meis1, and is most closely related to those of the Pbx/TGIF homeobox gene products. By in situ hybridization analysis, we show that the Meis2 gene displays spatially restricted expression patterns in the developing nervous system, limbs, face, and in various viscera. In adult mice, Meis2 is mainly expressed in the brain and female genital tract, with a different distribution of the alternative splice forms in these organs. PMID- 9337138 TI - Expression of Meis2, a Knotted-related murine homeobox gene, indicates a role in the differentiation of the forebrain and the somitic mesoderm. AB - Knotted (Kn) genes are expressed within restricted domains of the plant meristems and play a key role in the control of plant morphogenesis. We have isolated the Kn-related gene Meis2 in mouse, which labels the lateral somitic compartment and its derivatives during early mouse embryogenesis and later becomes a marker for the dorso-ectodermal region overlying cells of the paraxial mesoderm. Meis2 is also highly expressed in specific areas of the developing central nervous system from embryonic day 9 (e9) onward. In later developmental stages, a strong expression is detectable in differentiating nuclei and regions of the forebrain, midbrain, hindbrain, and spinal cord. This temporal and spatial expression pattern suggests that Meis2 may play an important role in the cascade of induction leading to somitic differentiation as well as in brain regionalization and histogenesis. PMID- 9337139 TI - Cellular and molecular bases of memory: synaptic and neuronal plasticity. AB - Discoveries made during the past decade have greatly improved our understanding of how the nervous system functions. This review article examines the relation between memory and the cellular mechanisms of neuronal and synaptic plasticity in the central nervous system. Evidence indicating that activity-dependent short- and long-term changes in strength of synaptic transmission are important for memory processes is examined. Focus is placed on one model of synaptic plasticity called long-term potentiation, and its similarities with memory processes are illustrated. Recent studies show that the regulation of synaptic strength is bidirectional (e.g., synaptic potentiation or depression). Mechanisms involving intracellular signaling pathways that regulate synaptic strength are described, and the specific roles of calcium, protein kinases, protein phosphatases, and retrograde messengers are emphasized. Evidence suggests that changes in synaptic ultrastructure, dendritic ultrastructure, and neuronal gene expression may also contribute to mechanisms of synaptic plasticity. Also discussed are recent findings about postsynaptic mechanisms that regulate short-term synaptic facilitation and neuronal burst-pattern activity, as well as evidence about the subcellular location (presynaptic or postsynaptic) of mechanisms involved in long term synaptic plasticity. PMID- 9337140 TI - Functional anatomy of long-term memory. AB - Memory in the brain is organized into multiple memory systems that perform different memory functions and have different neurologic substrates. Declarative memory involves conscious memory for facts and events. The medial temporal lobe and structures in the diencephalon are essential in the establishment of new declarative memories, and these memory traces are finally stored in domain specific regions of the cerebral cortex. The frontal lobe and basal ganglia are important in some forms of declarative memory that require reasoning about the contents of memory. Nondeclarative forms of memory (including skill learning, repetition priming, and classical conditioning) do not involve conscious recollection and are measured through changes in the way in which tasks are performed. These forms of memory rely upon the cerebral cortex, basal ganglia, and cerebellum. PMID- 9337142 TI - Prognostic value of EEG monitoring after status epilepticus: a prospective adult study. AB - Despite the significant morbidity and mortality associated with status epilepticus (SE), little is known about changes in cortical function that occur after SE. We evaluated cortical function after clinical SE using continuous EEG monitoring lasting at least 24 h in 180 patients admitted to the Medical College of Virginia Hospitals (MCVH). The major EEG patterns observed after SE were a normal record, burst suppression, after SE ictal discharge (ASIDs), periodic lateralizing epileptiform discharges (PLEDs), attenuation, focal and generalized slowing, and epileptiform discharges. Normalization of the EEG after SE was highly correlated with good outcome. The presence of burst suppression and ASIDs was highly statistically significantly associated with mortality. PLEDs were also highly correlated with mortality, but not to the same degree as burst suppression and ASIDs. In addition, these EEG patterns were still significantly correlated with morbidity and mortality when we controlled for etiology using multivariate logistic statistical analysis. Persistent ictal activity was observed in many patients despite control of clinical seizure activity, indicating the importance of EEG monitoring to determine treatment patterns after clinical seizure activity in SE is controlled. The results indicate that certain EEG patterns (normalization of the EEG, ASIDs, burst suppression and PLEDs) are useful predictors of outcome in SE in addition to etiology. EEG monitoring after control of clinical SE is important to guide treatment of SE and is a useful technique for evaluating prognosis. PMID- 9337141 TI - Use of the intracarotid amobarbital procedure in the evaluation of memory. AB - The intracarotid amobarbital procedure (IAP) involves the temporary inactivation of one cerebral hemisphere by the injection of sodium amobarbital, which allows independent testing of the contralateral hemisphere. Initially used for lateralization of language, IAP later found a role in the evaluation of memory function in patients with intractable temporal lobe epilepsy being considered for resective surgery. IAP technique varies widely across centers, but, in general, memory is assessed by presenting the patient with a number of items during the period of hemispheric inactivation and testing recall or recognition of these items after the effect of the drug has worn off. Because the medial temporal lobe is not directly perfused by the internal carotid artery, concerns have been raised about the ability of the IAP to test hippocampal memory function. Consequently, a variety of selective procedures have been devised. Findings on both intracranial EEG recordings and pathologic and neuroimaging studies support the association of IAP memory results with hippocampal function. The IAP memory test was originally designed to predict the risk for development of global amnesia following unilateral temporal lobectomy. More recently, it also has been used as an adjunct in lateralizing the seizure focus and for predicting postoperative selective memory deficits and seizure outcome. PMID- 9337143 TI - Fundamental electrophysiologic investigation of spinal cord: refractory period of feline conductive spinal cord evoked potential. AB - Refractory periods and recovery curves have been used to investigate the physiologic importance and disturbance of peripheral nerves, but the refractory periods of the central nervous system (CNS) have seldom been investigated. We estimated the refractory periods and the recovery curves of the ascending and descending conductive spinal cord evoked potentials (SCEP) in cats. The absolute refractory period of the first and second potentials of both the ascending and descending SCEP was approximately 0.4-0.5 ms. The amplitudes of the first potentials of the ascending and descending SCEP elicited by test stimuli exhibited significant differences, but their latencies did not differ significantly except at the interstimulus interval (ISI) of 1.5 ms, which implies that the same type of fibers was stimulated in the first potentials of the ascending and descending SCEP. The second potential of the descending SCEP elicited by test stimulus showed > 100% amplitude and a maximal recovery of 200% when the ISI was 3.0 ms. The third potential was produced in the test response more easily when a lower vertebral level (L4) was used as the recording site and the ISI was between 1.0 and 4.0 ms. We consider these phenomena to be the result of elimination of the synaptic inhibitory influence by the conditioning stimulus of the paired stimuli for the descending SCEP. PMID- 9337144 TI - Monitoring changes in the health of the U.S. elderly population: correlates with biomedical research and clinical innovations. PMID- 9337145 TI - Adriamycin cardiomyopathy: pathophysiology and prevention. AB - Current knowledge about adriamycin cardiomyopathy indicates that the major cause of this condition is increased oxidative stress although the drug's antitumor action in patients may involve other mechanisms. Controversies about the different antioxidants in preventing cardiomyopathy likely stem from the fact that antioxidants must be effective in both the lipid and water phases, and the dose must be optimal, in order to be protective. Probucol, an antioxidant and promoter of endogenous antioxidants, is one such agent. Conducting clinical trials with an optimal dose of probucol is the next step and should make this great anticancer drug safer and more efficient in the fight against the cancer. PMID- 9337146 TI - Genetics of human obesity: research directions. AB - Rapid strides in understanding the physiology controlling energy or nutrient intake and energy expenditure have complemented the search for the genetic basis of obesity. Several single gene defects are known that produce obesity in animals. All of these have been cloned within the past 4 years, providing a rich new base for understanding obesity. Since obesity is likely to be "multifactorial," a number of laboratories have used the quantitative trait locus (QTL) technique of genome scanning to identify candidate genomic regions and, eventually, genes that may influence body weight and body fat. So far, 18 QTLs have been identified in association with crossbreeding strains of mice or rats with variable susceptibility to obesity. A number of mendelian disorders are known to exist in humans, but no specific genes have yet been identified for them. The potential for inserting new genetic material into mammals has produced numerous transgenic mice with increased or decreased quantities of body fat. These models will provide a continuing source of new insights into obesity. Several areas in the human genome have been linked to the development of obesity. Among the candidate genes with evidence of linkage to body fat are TNF-alpha, adenosine deaminase, and melanocortin-3 receptor. The new insights described above have invigorated the pharmaceutical industry to increase their efforts for new drug development aimed at the growing problem of obesity. PMID- 9337147 TI - Bcl-2 overexpression enhances the metastatic potential of a human breast cancer line. AB - Bcl-2 protein has been shown to contribute to oncogenesis because it can transform and immortalize cells in cooperation with c-myc, ras, or viral genes. However, in vivo studies have not yet established whether bcl-2 can play a role in metastasis. Here we investigate the potential metastatic role of bcl-2. We introduced the human bcl-2 gene into a low bcl-2 expressing human breast cancer cell line MCF7 ADR. We demonstrate that two bcl-2 overexpressing clones injected intravenously or intramuscularly into nude mice induce a significantly higher number of experimental and spontaneous lung metastases compared to the control transfectant clone. We demonstrate that bcl-2 overexpressing clones are more invasive and migratory in response to chemotactic stimuli than the control transfectant clone. Furthermore, zymographic analysis shows that secretion of 72 and 92 kDa gelatinases increases in the two bcl-2 overexpressing transfectants. Tumors originating from bcl-2 overexpressing clones also show a decrease in the latency period of tumor appearance. In conclusion, our data show that bcl-2 overexpression enhances both tumorigenicity and metastatic potential of MCF7 ADR cells by inducing metastasis-associated properties. PMID- 9337148 TI - Inducible nitric oxide synthase-deficient mice have enhanced leukocyte endothelium interactions in endotoxemia. AB - Nitric oxide (NO) from constitutive NO synthase (NOS) has been postulated to be a homeostatic regulator of leukocyte-endothelial cell interactions. By contrast, the inducible NO synthase (iNOS) isoform has been invoked as a potential pathogenic enzyme in numerous inflammatory diseases. The objective of this study was to determine whether the iNOS isoform is also capable of functioning as a regulator of leukocyte recruitment. Mice received endotoxin (LPS, 30 microg/kg, i.v.); 2-4 h later, intravital microscopy was used to examine leukocyte rolling and adhesion in postcapillary venules of the cremaster muscle and the sinusoids and postsinusoidal venules of the hepatic microcirculation. Leukocyte recruitment into the lung was also examined. RT-PCR confirmed that this treatment induced iNOS mRNA expression in wild-type mice as early as 2 h after LPS treatment. Between 2 and 4 h after LPS administration, the number of rolling and adherent leukocytes in cremasteric postcapillary venules and of adherent cells in liver postsinusoidal venules of iNOS-deficient mice were significantly higher than in wild-type mice. Leukocyte accumulation in the lung (measured by myeloperoxidase assay) was also significantly elevated in iNOS-deficient animals. These effects could not be attributed to differences in systemic blood pressure, shear rates, circulating leukocyte numbers, or baseline levels of rolling and adhesion because these parameters were not different between the two groups. To establish whether the differences in leukocyte recruitment were related to the leukocytes per se, perfusion of iNOS+/+ or iNOS-/- septic blood over purified E-selectin (using parallel plate flow chambers) revealed much larger recruitment of iNOS-/- leukocytes. These results suggest that iNOS induced in response to LPS releases NO that is capable of reducing leukocyte accumulation by affecting leukocytes directly and raises the possibility that induction of iNOS is a homeostatic regulator for leukocyte recruitment. PMID- 9337149 TI - The activated anaplastic lymphoma kinase increases cellular proliferation and oncogene up-regulation in rat 1a fibroblasts. AB - More than 60% of anaplastic large-cell lymphomas (Ki-1 lymphoma) are associated with a t(2;5)(p23;q35) translocation that produces an 80 kDa hyperphosphorylated chimeric protein (p80) derived from the fusion of the anaplastic lymphoma kinase (ALK) with nucleophosmin (NPM). The NPM-ALK chimeric gene is an activated tyrosine kinase that has been shown to be a potent oncogene. We have developed a cellular model for the study of p80 action in rat 1a fibroblasts. Expression of cDNA's encoding NPM-ALK (p80) in rat 1a fibroblasts induces anchorage-independent growth in soft agar and promotes foci formation in culture. Cells expressing exogenous p80 showed significantly increased proliferation characterized by accelerated cell cycle entry into S-phase. Consistent with increased G0/G1 to S phase transition, there is also marked up-regulation of cyclin A and cyclin D1 expression. In addition, p80 transformed cells showed elevated expression of several immediate early genes involved in cellular proliferation, including fos, jun, and c-myc. DNA binding analysis of nuclear extracts prepared from p80 transformed cells reveal marked up-regulation of AP-1 DNA binding activity. Functional AP-1-specific transfection assays also show up-regulation of AP-1 dependent transcriptional activation. These finding demonstrate that p80 transformed rat 1a fibroblast can be a highly useful model system for the molecular and biochemical characterization of the mechanisms of action of this interesting new oncogene. PMID- 9337150 TI - Epinephrine enhances glycogen turnover and depresses glucose uptake in vivo in rat heart. AB - In vivo effects of epinephrine on glucose uptake and glycogen turnover in rat heart were studied and compared to liver and skeletal muscle. Fasted ketamine anesthetized rats were intravenously infused with saline or epinephrine. Both the low and high doses of epinephrine resulted in hyperglycemia (40-50%) and hyperlactemia (threefold) at the end of infusion. Glucose uptake, determined by the phosphorylation of the intravenously injected [14C]2-deoxyglucose, was found to decrease in the heart and skeletal muscle of epinephrine-infused rats. Glycogen in livers, skeletal muscles, and hearts of the epinephrine-infused rats decreased to varying degrees relative to the saline-infused rats, indicating enhanced glycogenolysis in all three organs. Glycogen synthesis, determined by the incorporation of the co-infused [3-(3)H]glucose into glycogen, was found to decrease in liver and skeletal muscle. However, glycogen synthesis in the heart was found to increase 50% in Epi-1 and 280% in Epi-2 compared to the saline infused rats. We conclude that glucose utilization in the in vivo heart may be preferentially channeled through glycogen turnover in the presence of epinephrine. That both synthesis and degradation of glycogen can be simultaneously activated appears to be unique to the heart and is protective against a loss of glycogen at a time of enhanced glucose utilization. PMID- 9337151 TI - Characterization of diadenosine polyphosphate transport into chromaffin granules from adrenal medulla. AB - The transport of diadenosine polyphosphates into chromaffin granules from bovine adrenal medulla has been studied by using the radiolabeled substrate [3H]Ap5A and the fluorescent substrate analog di(1,N6-ethenoadenosine)polyphosphate, epsilon (Ap(n)A) (n=3-5). The vesicular concentration increase was time dependent and the substrates were not metabolized to any extent during the transport experiments. The saturation curve indicates the existence of kinetic and allosteric cooperativity during Ap(n)A (diadenosine polyphosphates) transport and could be the result of the presence of various affinity states of the transporter with K values of 16 +/- 1 microM and 75 +/- 6 microM, and corresponding Hill numbers of 2 and 4, when epsilon-(Ap4A) was the substrate. The saturation studies for [3H]Ap5A were performed in a broader concentration range; in this case a three step curve was obtained with K values of 16 +/- 2 microM, 125 +/- 9 microM, and 545 +/- 11 microM; the corresponding Hill numbers were 2, 4, and 6. This kinetic behavior can be explained on the basis of a mnemonic model, as already demonstrated for the vesicular transport of ATP. The nonhydrolyzable adenine nucleotide analogs, ATPgammaS and ADPbetaS, inhibited the diadenosine polyphosphate transport at concentrations in the millimolar range. Ap(n)A transport was also inhibited by the P2 receptor antagonist suramin, the mitochondrial ATP/ADP exchange inhibitor atractyloside, the proton translocator FCCP, and N-ethylmaleimide. PMID- 9337152 TI - Latent transforming growth factor beta1 activation in situ: quantitative and functional evidence after low-dose gamma-irradiation. AB - The biological activity of transforming growth factor beta1 (TGF-beta) is controlled by its secretion as a latent complex in which it is noncovalently associated with latency-associated peptide (LAP). Activation is the extracellular process in which TGF-beta is released from LAP, and is considered to be a primary regulatory control. We recently reported rapid and persistent changes in TGF-beta immunoreactivity in conjunction with extracellular matrix remodeling in gamma irradiated mouse mammary gland. Our hypothesis is that these specific changes in immunoreactivity are indicative of latent TGF-beta activation. In the present study, we determined the radiation dose response and tested whether a functional relationship exists between radiation-induced TGF-beta and collagen type III remodeling. After radiation exposures as low as 0.1 Gy, we detected increased TGF beta immunoreactivity in the mammary epithelium concomitant with decreased LAP immunostaining, which are events consistent with activation. Quantitative image analysis demonstrated a significant (P=0.0005) response at 0.1 Gy without an apparent threshold and a linear dose response to 5 Gy. However, in the adipose stroma, loss of LAP demonstrated a qualitative threshold at 0.5 Gy. Loss of LAP paralleled induction of collagen III immunoreactivity in this tissue compartment. We tested whether TGF-beta mediates collagen III expression by treating animals with TGF-beta panspecific monoclonal antibody, 1D11.16, administered i.p. shortly before irradiation. Radiation-induced collagen III staining in the adipose stroma was blocked in an antibody dose-dependent manner, which persisted through 7 days postirradiation. RNase protection assay revealed that radiation-induced elevation of total gland collagen III mRNA was also blocked by neutralizing antibody treatment. These data provide functional confirmation of the hypothesis that radiation exposure leads to latent TGF-beta activation, support our interpretation of the reciprocal shift in immunoreactivity as evidence of activation, and implicate TGF-beta as a mediator of tissue response to ionizing radiation. The sensitivity of activation to low radiation doses points to a potential role for TGF-beta in orchestrating tissue response to oxidative stress. As such, radiation may be useful as a probe to delineate the consequences of latent TGF-beta activation in situ. PMID- 9337153 TI - CD10 is expressed on human thymic epithelial cell lines and modulates thymopentin induced cell proliferation. AB - Thymic hormones such as thymopoietin (TP) have been shown to regulate thymocyte differentiation and lymphocyte activation. However, it is not known whether thymopoietin affects thymic epithelial cell (TEC) functions. In this study we have examined the effect of a five amino acid active peptide (TP5), corresponding to amino acids 32-36 of TP, on the proliferation of nontransformed clones of human TEC. Our results indicate that TP5 induced reinitiation of DNA synthesis and potentiated fetal calf serum (FCS)-induced cell growth in postnatal and fetal derived human TEC. We also found that TEC lines express high levels of endopeptidase 24.11, a cell-surface metallopeptidase also known as the CD10 antigen. We show that TP5 is cleaved by CD10 at the surface of TEC lines, indicating that this endopeptidase may regulate TP5-induced TEC proliferation. Phosphoramidon, a specific endopeptidase 24.11 inhibitor, consistently acts in synergy with TP5 to enhance FCS-induced TEC growth. Hence, we conclude that 1) TP5 alone or in combination with FCS supports the growth of TEC lines, and 2) TEC lines express high levels of CD10, which regulates TP5-induced TEC proliferation by acting as a thymic peptide degrading enzyme. PMID- 9337154 TI - Interaction of liver methionine adenosyltransferase with hydroxyl radical. AB - Liver methionine adenosyltransferase (MAT) plays a critical role in the metabolism of methionine converting this amino acid, in the presence of ATP, into S-adenosylmethionine. Here we report that hydrogen peroxide (H2O2), via generation of hydroxyl radical, inactivates liver MAT by reversibly and covalently oxidizing an enzyme site. In vitro studies using pure liver recombinant enzyme and mutants of MAT, where each of the 10 cysteine residues of the enzyme subunit were individually changed to serine by site-directed mutagenesis, identified cysteine 121 as the site of molecular interaction between H2O2 and liver MAT. Cysteine 121 is specific to the hepatic enzyme and is localized at a "flexible loop" over the active site cleft of MAT. In vivo studies, using wild-type Chinese hamster ovary (CHO) cells and CHO cells stably expressing liver MAT, demonstrate that the inactivation of MAT by H2O2 is specific to the hepatic enzyme, resulting from the modification of the cysteine residue 121, and that this effect is mediated by the generation of the hydroxyl radical. Our results suggest that H2O2-induced MAT inactivation might be the cause of reduced MAT activity and abnormal methionine metabolism observed in patients with alcoholic liver disease. PMID- 9337155 TI - Molecular epidemiology of atherosclerosis. AB - It has been hypothesized that mutational events may be involved in the atherogenetic process and that at least a portion of atherosclerotic plaques may develop according to an initiation-promotion process of arterial smooth muscle cells, akin to benign tumors. We conducted a study to evaluate the occurrence of oxidative DNA damage and formation of DNA adducts in human atherosclerotic lesions and to assess the relationships of these promutagenic alterations with exposure to atherogenic risk factors. Pure DNA was extracted from the tunica media (composed mainly of smooth muscle cells) of abdominal aorta fragments taken at surgery from 85 patients suffering from severe atherosclerotic lesions. DNA adducts were detected by synchronous fluorescence spectrophotometry and 32P postlabeling after enrichment of adducts with either butanol or nuclease P1. 8 Hydroxy-2'-deoxyguanosine (8-OH-dG), a typical indicator of oxidative DNA damage, was measured by HPLC/electrochemical detection. A complete questionnaire reporting general, clinical, and laboratory characteristics was available for each patient. All 84 samples tested by 32P postlabeling were positive by displaying the presence of diagonal radioactive zones and up to 9 individual DNA adducts. Of 52 samples tested, 32 (61.5%) yielded typical positive signals at synchronous fluorescence spectrophotometry. All but one of 39 samples tested had very high levels of 8-OH-dG, thus showing a remarkable oxidative DNA damage. Statistically significant correlations were found between the levels of molecular biomarkers and atherogenic risk factors including age, number of currently smoked cigarettes, ratio of total-to-high density lipoprotein blood cholesterol, blood triglycerides, and blood pressure. The DNA alterations detected in our study may be only one component of the genetic basis of atherogenesis. Moreover, no causal role in the atherogenetic process can be inferred from our results. However, DNA alterations, including oxidative damage and adduction of reactive molecules of either endogenous or exogenous source, were systematically present in the smooth muscle cells of human atherosclerotic lesions and their intensity was significantly correlated with the occurrence of atherogenic risk factors in the patients studied. PMID- 9337156 TI - Development of pathogenic anti-DNA antibodies in patients with systemic lupus erythematosus. AB - The anti-DNA response is a hallmark of systemic lupus erythematosus (SLE). The precise mechanisms leading to anti-DNA antibody (Ab) production remain to be studied. Nonetheless, it is becoming clear that anti-DNA Abs cause inflammatory lesions not only via deposition of circulating immune complexes (IC) consisting of anti-DNA Ab and antigens (Ags), but also via in situ IC formation by cationic anti-DNA Abs. It is intriguing that cationic anti-DNA Abs are encoded by a unique germline Vkappa gene, A30, which encodes an extraordinary cationic light chain, whereas somatic mutations did not induce a cationic shift of electrical charge in human lupus nephritis, suggesting that the usage of a specific germline gene may confer the cationic charge (or pathogenicity) on anti-DNA Abs and that somatic mutations induce the affinity maturation of Abs. Whether cationic anti-DNA Abs will develop depends at least partly on the presence or absence of the germline A30 gene, since patients who lack this gene in the germline Vkappa repertoire did not develop severe lupus nephritis. Receptor editing, a mechanism for changing the affinity of the B cell Ag receptor [surface immunoglobulin (Ig) receptor] to avoid self-reactivity actually seems defective in patients with SLE because normal B cells edited the A30 gene, whereas SLE B cells express A30 mRNA. Thus, along with the importance of somatic mutations, polymorphisms of Ig Vkappa locus, and genetic predisposition, the failure of receptor editing may contribute to the development of pathogenic anti-DNA responses in humans. PMID- 9337157 TI - The uniform approach to breast fine-needle aspiration biopsy. NIH Consensus Development Conference. PMID- 9337158 TI - Consensus statement on fine-needle aspiration. PMID- 9337159 TI - Effect of duodenum-preserving resection of the head of the pancreas on endocrine and exocrine pancreatic function in patients with chronic pancreatitis. AB - BACKGROUND: Chronic pancreatitis leads to progressive destruction of pancreatic parenchyma affecting exocrine and endocrine function. We prospectively evaluated the effect of duodenum-preserving resection of the head of the pancreas on pancreatic function. METHODS: Exocrine and endocrine function were measured in a combined test including (1) urinary PABA recovery; (2) plasma glucose, glucagon, and C-peptide responses; and (3) plasma pancreatic polypeptide response. Nineteen patients were included. RESULTS: Compared with the preoperative state, plasma glucose levels did not increase postoperatively. Plasma C-peptide levels were reduced postoperatively but the difference was not significant. The percentage of insulin-dependent patients did not increase after operation (32% versus 32%). Glucose tolerance improved in 4 patients and deteriorated in 3 patients. Postoperative basal and-meal stimulated plasma pancreatic polypeptide levels were significantly reduced. Postoperative urinary PABA recovery was not significantly different from preoperative values. CONCLUSIONS: Neither exocrine nor endocrine pancreatic function are negatively influenced by duodenum-preserving pancreatic head resection. PMID- 9337160 TI - Pancreaticoduodenectomy for nonperiampullary primary tumors. AB - INTRODUCTION: This review was performed to evaluate the outcome of patients undergoing pancreaticoduodenectomy (PD) for isolated metastatic or locally advanced nonperiampullary tumors at a single institution over a 13-year period. METHODS: Between 1983 and 1996, patients undergoing PD for metastatic or locally advanced nonperiampullary malignancies were identified. Medical records were reviewed and outcome factors and survival data analyzed. RESULTS: Eighteen patients were identified. The primary tumor histopathology included colon (n = 7), gastric (n = 4), renal cell (n = 3), lung (n = 2), bladder (n = 1), and melanoma (n = 1). The median length of hospital stay was 15 days (6 to 48) with one perioperative death (5.5%). The median tumor size was 5.5 cm (0.8 to 11.5), and 7 patients had positive peripancreatic lymph nodes. The median survival was 40 months, with a 5-year survival of 35%. CONCLUSIONS: Pancreaticoduodenectomy for nonperiampullary malignancy is infrequently indicated. However, in the absence of widely metastatic disease, PD should be considered for locally advanced tumors or isolated metastatic malignancy. PMID- 9337161 TI - Spiral computed tomography scanning after intravenous infusion cholangiography for biliary duct anomalies. AB - BACKGROUND: Iatrogenic injury of the bile duct during cholecystectomy represents a failure of surgical technique, especially for laparoscopic surgery. Knowledge of the patient's individual ductal anatomy and anomalies preoperatively would be helpful in avoiding such injuries. Therefore, we investigated the anatomy of the biliary duct and any anomalies using spiral computed tomography (SCT) scanning following intravenous infusion cholangiography (IVC-SCT). MATERIALS: Laparoscopic cholecystectomies (LC) were attempted on 437 patients at the Kansai Medical University. Preoperative IVC-SCT and laparoscopic cholangiography were attempted in all of the patients. RESULTS: An overall anomalous union of the cystic duct was seen in 71 (16.2%) out of the 437 patients subjected to IVC-SCT. The following anomalies were observed: right hepatic duct entry in 7 cases (1.6%), parallel low entry in 17 cases (3.9%), posterior spiral entry in 35 cases (8.0%), anterior spiral entry in 7 cases (1.6%), and accessory duct entry in 5 cases (1.1%). The success rate for the LC was 99.5% (435/437). Three patients were switched to open surgery owing to advanced gallbladder cancer and severe adhesions. The success rate for the laparoscopic cholangiography was 97.2% (423 of 435). Intraoperative right hepatic duct injury occurred in only 1 patient with a bile duct anomaly, and it was repaired with laparoscopic T-tube drainage. CONCLUSIONS: The preoperative examination of the biliary tract by IVC-SCT was technically simple, less invasive, and may helpful in avoiding damage to the bile duct, especially in patients with biliary duct anomalies. PMID- 9337162 TI - Intraoperative events common to videoscopic preperitoneal mesh inguinal herniorrhaphy. AB - BACKGROUND: Videoscopic preperitoneal mesh (VPM) inguinal herniorrhaphy avoids the entry into the abdominal cavity, which is necessary with other videoscopic techniques. Despite this advantage, surgeons have been slow to adopt this technique. We reviewed our experience with VPM inguinal herniorrhaphy, specifically investigating the technical aspects of this approach. METHODS: Data were collected prospectively. Operative notes were reviewed retrospectively detailing intraoperative events not "typical" with the VPM technique. RESULTS: One hundred consecutive patients undergoing VPM repair of 127 hernias were studied. The repair was completed in all but 2 patients. Mean operating time was 120 minutes (60 to 146). In 36 repairs there were 59 intraoperative "events" requiring specific maneuvers to correct. Events identified were the need for transection of the hernia sac, creation and repair of a peritoneal tear, and need to divide the inferior epigastric vessels. No complications related to these events occurred. When events occurred, operative times were significantly longer (146+/-45 versus 83+/-23 minutes; P <0.05). CONCLUSION: Intraoperative events are common with VPM herniorrhaphy. These events significantly prolong operating time. A surgeon's lack of familiarity with such events and how to deal with them may in part explain the reluctance to widely apply the VPM technique. PMID- 9337163 TI - Voice activation of a surgical robotic assistant. PMID- 9337164 TI - Objective assessment of endoscopic knot quality. AB - BACKGROUND: Studies of the surgeon's skill and the ergonomics of task performance in endoscopic surgery can be based on knot-tying tasks. The aim of this study was to establish an objective method for assessing the quality of surgical knots for use in such studies. METHODS: In all, 2,700 surgeon's endoscopic knots were studied. Each knot was distracted using a tensiometer, and a computerized system analyzed force-extension curves. The breaking force was taken as an index of knot strength while the force integrated over the slope of the curve reflected knot tightening. A knot quality score (KQS) was obtained from the product of the knot breaking force and the integrated force expressed as a percentage of the product for the untied ligature. RESULTS: The mean breaking force (24 Newton +/- 2.5) and integrated force (7.4 Newton +/- 2.8) for broken knots were 71% and 35%, respectively, of those for untied ligature. The integrated force yielded a narrower range of variability for untied ligature (SD 3.5% of mean) than for knots (SD 37% of mean). The KQS was higher for broken (25.3%+/-10.3%) than slipped knots (7.1%+/-5.1%). CONCLUSION: The KQS provides a reliable assessment of knot security and reflects the strength and degree of tightening of the knot. PMID- 9337165 TI - Surgical therapy for 101 patients with acquired immunodeficiency syndrome and symptomatic cholecystitis. AB - BACKGROUND: Hepatobiliary disease in patients with acquired immunodeficiency syndrome (AIDS) has been well documented. Cytomegalovirus and Cryptosporidium are the pathogens most frequently associated. Previous reports of cholecystectomies and AIDS have had conflicting results on morbidity and mortality. METHOD: Retrospective review of 101 patients with AIDS and symptomatic cholecystitis who underwent cholecystectomy from December 1989 to May 1995. RESULTS: All patients had symptoms characteristic of gallbladder disease, the most common being abdominal pain and fever. Thickening of the gallbladder was the most common diagnostic finding. Fifty-six patients underwent open cholecystectomy and 45 laparoscopic cholecystectomy. Pathologic examination revealed an abnormal gallbladder in all cases and gallstones in 29%. A specific pathogen or malignancy was identified as the etiologic agent in 44% of patients. Perioperative morbidity was similar (<5%) in both surgical groups. Perioperative mortality was 4% among all the patients treated. CONCLUSIONS: Both open and laparoscopic cholecystectomy improved the quality of life of these patients and should be considered as the treatment for persistent hepatobiliary symptoms in patients with AIDS. PMID- 9337166 TI - The timing and sequence of multiple device-related complications in patients with indwelling subcutaneous ports. AB - BACKGROUND: Multiple complications associated with venous access ports are a common occurrence. In an effort to define patterns of sequential complications in our community, we undertook a prospective analysis of adult cancer patients in whom a subcutaneous port was inserted. METHODS: One hundred nineteen consecutive adult cancer patients in whom a subcutaneous port was inserted were observed prospectively for the development of complications. RESULTS: Complications were identified in 70 of the 91 evaluable patients, while sequential complications were identified in 35 patients (38%). In aggregate, 121 complications were identified. The ball-valve effect, the most frequently identified problem, was found to occur disproportionately as a primary complication (52 of 70 versus 26 of 51, P <0.02). In contrast, port-related venous thrombosis was identified most frequently as a subsequent complication (11 of 51 versus 4 of 70, P <0.02). The only identified risk factor for the development of port-related complications was the ball-valve effect, found to be associated with the subsequent development of port-related venous thrombosis (9 of 52 versus 2 of 69, P <0.02). CONCLUSIONS: Multiple sequential complications of subcutaneous ports are common and occur in a rather predictable order. The occurrence of port-related venous thrombosis in patients with an earlier, relatively minor vascular complication (ball-valve effect) suggests a cause-effect relationship. Insight into complication sequencing may lead to improved strategies for prevention and therapy. PMID- 9337167 TI - Preventing infection of the incision after appendectomy by using metronidazole preoperatively to infiltrate tissues at the incision. AB - METHODS: From January 1991 to July 1995, 260 patients with acute appendicitis were operated on. After excluding 21 patients, the remaining 239 cases were randomly divided into two groups. Group A was given the treatment and group B was the control group. Precisely 0.915 g metronidazole disodium phosphate injection (Tongzhen Pharmaceutical Co., Shan Xi Province, P. R. China) or 25% metronidazole glucose solution was added to 100 mL 0.9% normal saline. After anesthetizing the patients in group A, 60 to 80 mL of the solution was injected into the subcutaneous tissue and muscle. The control group B was given intravenous injection of metronidazole disodium phosphate and cephazolin postoperatively. RESULTS: In the 119 cases in group A, the rate of incision infection was 0.8%. By contrast, the rate of infection among the 120 cases in group B was 11.6%. The statistical analysis (chi square) was significant, with P < 0.001 showing that preincisional or intraincisional infiltration with metronidazole was effective. PMID- 9337168 TI - Proliferating cell nuclear antigen as a marker of cell kinetics in aberrant crypt foci, hyperplastic polyps, adenomas, and adenocarcinomas of the human colon. AB - BACKGROUND: One of the first steps in multistage colonic carcinogenesis is increased cell proliferation and an upward shift of the proliferation zone of colonic crypts. In the present study, progression in cell kinetics was followed up at all sequential stages of colonic carcinogenesis, starting with aberrant crypt foci (ACF), the earliest putative preneoplastic lesions, hyperplastic and dysplastic polyps, and invasive carcinomas. MATERIALS AND METHODS: Colonic tissue and tumor specimens were prospectively obtained from 65 patients treated at our hospital for adenocarcinoma or malignant polyps. For identification of ACFs, dissected mucosal strips obtained from patients with colorectal cancer were stained with 0.1% methylene blue and scanned under dissecting microscope. Paraffin-embedded ACFs and macroscopic lesions were serially sectioned, deparaffinized, and stained with a monoclonal antiproliferating cell nuclear antigen (PCNA) antibody. The PCNA-labelling index (PCNA-LI), expressed as a ratio of positively stained nuclei to total nuclei counted, was calculated separately for basal, middle, and upper colonic crypt compartments. A comparison of the PCNA LI was made for each compartment in normal mucosa, and hyperplastic and dysplastic lesions. RESULTS: A stepwise increase in the PCNA-LI was observed during neoplastic progression of colonic lesions. The two most important variables of increased cell proliferation, expressed as PCNA-LI per crypt compartment, were the presence of dysplasia and the size of dysplastic lesions. CONCLUSIONS: In colorectal carcinogenesis, hyperproliferation with upward expansion of proliferative compartment is a characteristic feature at all stages of malignant progression. PMID- 9337169 TI - High prevalence of bone disorders after gastrectomy. AB - BACKGROUND: Studies indicate that gastrectomy might alter calcium and bone metabolism, resulting in bone disorders. No data are currently available on the prevalence of bone disorders after gastrectomy. METHODS: Sixty gastrectomy patients were investigated for serum parameters of calcium and bone metabolism 5 to 20 years postoperatively and compared to an age- and sex-matched healthy control population. Forty patients agreed to a radiological investigation of the spine by anterior-posterior and lateral radiographs of the thoracic and lumbar spine and by computed tomography (CT) osteodensitometry. RESULTS: Serum calcium and 25-(OH)-vitamin D were decreased in gastrectomized patients, while parathyroid hormone and 1,25-(OH)2-vitamin D were increased. Serum parameters of calcium metabolism were altered in as many as 68% of patients. We found 31 vertebral fractures in 13 patients, 30 grade 2 vertebral deformities in 18 patients, and osteopenia in 15 patients, corresponding to a prevalence of 33%, 45%, and 37% in gastrectomized patients, respectively. The overall rate of gastrectomy patients having vertebral fractures and/or osteopenia was 55%. The risk of having a vertebral deformity was increased by more than sixfold after gastrectomy. Our study is the first report evaluating vertebral deformities in gastrectomized patients, and the largest series of gastrectomized patients investigated by CT osteodensitometry. CONCLUSION: We found a high prevalence of bone disorders in gastrectomized patients, possibly resulting from disorders in calcium metabolism. Postgastrectomy bone disease might derive from a calcium deficit, which increases calcium release from bone and impairs calcification of newly build bone matrix. PMID- 9337170 TI - Surgical debridement and intralesional steroid injection in the treatment of idiopathic AIDS-related anal ulcerations. AB - BACKGROUND: The treatment of idiopathic anal ulcerations in AIDS patients is still evolving. These patients suffer with severe incapacitating anal pain. PATIENTS AND METHODS: Relief of pain was achieved with the use of intralesional steroid injections. Twenty-one patients from 1990 to 1993 presented with severe anal pain of 4.5 months average durations (range 3 weeks to 2 years). The average CD4 count was 52.1 (range 0 to 150). Fourteen (67%) patients had a solitary ulcer, while 7 (33%) had complex ulcerations. RESULTS: All patients were treated by debridement of the ulcer, biopsy, and intralesional injection of steroids. Microscopic evaluation revealed nonspecific inflammatory changes, without histopathologic evidence of viral infection or malignancy. Viral tissue cultures were negative. Fifty-seven percent of patients required only one injection to achieve dramatic relief of pain. Forty-three percent necessitated additional injections at 2-week intervals. The average patient was injected 1.9 times (range 1 to 7), with 20 of 21 patients (95%) reporting good to excellent pain relief. CONCLUSION: AIDS anal ulcerations should be treated aggressively with surgical debridement, biopsy and intralesional steroids. The efficacy of this therapy suggests that symptoms are partially due to inflammatory mediators. PMID- 9337171 TI - Endoscopic dexamethasone injection following balloon dilatation of anastomotic stricture after esophagogastrostomy. AB - BACKGROUND: Anastomotic stricture is common after esophagogastrostomy. Recent advances in nonsurgical treatment include the silicon bougie and balloon dilatation. However, simple dilatation alone with a silicon bougie or endoscopic balloon dilator was repeated a mean of 4.7+/-5.4 times to control anastomotic stricture because of its temporary effect. METHODS: For 11 patients, endoscopic injection of dexamethasone (8 mg) around the anastomosis was done immediately after balloon dilatation (40 psi for 5 minutes). RESULTS: This method significantly reduced the number of the dilatations to 1.1+/-0.3 (P < 0.05). Ten of the 11 patients did not need any further treatment. There were no side effects or complications of dexamethasone injection. CONCLUSION: A combination of endoscopic balloon dilatation and dexamethasone injection provided an easy and safe method for preventing the recurrence of anastomotic stricture. PMID- 9337172 TI - Inflammatory response after abdominal trauma, infection, or intestinal obstruction measured by oxygen radical production in peritoneal fluid. AB - BACKGROUND: Reactive oxygen intermediates (ROI) have been implicated in many pathophysiological processes of inflammatory tissue damage and tissue repair. In the present study we compared the production of ROI in three different types of tissue damage in surgical patients. METHODS: Peritoneal fluid specimens were harvested during the initial operation and postoperatively from 25 surgical patients with abdominal trauma, intraabdominal infection, and intestinal obstruction. The optical density at 412 nm, representing the peroxidation of hemoglobin, was measured to assess intraperitoneal ROI production. Patients were categorized into 3 groups: (A) infected patients with good outcome, (B) patients after trauma or obstruction with good outcome, and (C) patients with poor outcome due to persistent or secondary infection and multiple organ failure. Analysis of variance (ANOVA) and paired t test were used for statistical analysis. RESULTS: Overall, the ROI production decreased significantly at days 2 and 3 compared with day 0 and 1 (P = 0.0013). No initial differences of intraoperative ROI concentrations were found among the three groups; however, patients with a poor outcome showed increased ROI values after 4 to 5 days (P = 0.038) when compared with the good outcome group. CONCLUSIONS: We have demonstrated that intraperitoneal ROI production (1) can be measured in patients with intraabdominal tissue damage, (2) is not different between patients with intraabdominal infections, abdominal trauma, or intestinal obstruction, and (3) correlates with the clinical picture and the presence of an inflammatory intraabdominal focus or tissue damage. PMID- 9337173 TI - Magnetic resonance-guided laparoscopic interstitial laser therapy of the liver. AB - BACKGROUND: There is a necessity for an imaging method during laparoscopy to get a three-dimensional access to the target. In this study we evaluated laparoscopic interstitial laser therapy of the liver under magnetic resonance imaging guidance. METHODS: Five domestic pigs underwent laparoscopy in an open configuration magnetic resonance system. Under simultaneous real-time magnetic resonance imaging interstitial laser therapy was applied to the liver. Magnetic resonance images, macroscopic aspects of the lesions, and light microscopic findings were compared. RESULTS: The interventions could be safely performed. There was no image artifact caused by instruments or by the carbon dioxide. Dynamic gadolinium-enhanced imaging proved to significantly predict the macroscopic volume of the laser lesions. CONCLUSIONS: Magnetic resonance-guided laparoscopic interstitial laser therapy of the liver combines the advantages of minimal invasive surgery and magnetic resonance imaging. Further development should focus on laparoscopic instruments and temperature sensitive sequences. PMID- 9337174 TI - What surgeons need to know to help patients receive quality medical care in managed care settings. AB - BACKGROUND: Americans spend more than $3,300 per person per year on health care, an amount businesses, government, and citizens alike consider too high. MATERIAL REVIEWED: Managed care attempts to offer services at a lower cost, about 10% below that of indemnity insurance, and attempts to controls costs by modifying decisions historically made by physician and patient. Many techniques have been used to modify the decision-making process, with varying effects on quality. CONCLUSIONS: Surgeons can help sustain easily accessible, high-quality care through various personal behaviors and through choosing their managed care partners well. PMID- 9337175 TI - Benefits of fibrate drugs in coronary heart disease patients with normal cholesterol levels. PMID- 9337176 TI - Inflammation, metalloproteinases, and increased proteolysis: an emerging pathophysiological paradigm in aortic aneurysm. PMID- 9337177 TI - Direct thrombin inhibition superior to heparin during and after thrombolysis: dose, duration, and drug. PMID- 9337178 TI - Is it important how one dies? Questions for planning future revascularization trials. PMID- 9337179 TI - Intravascular ultrasound-guided percutaneous fenestration of the intimal flap in the dissected aorta. AB - BACKGROUND: Aortic dissection with branch obstruction is associated with high morbidity and mortality. Fenestration of the dissection flap to relieve distal vessel ischemia is at present largely performed surgically. The surgical mortality and morbidity are high, because most patients are poor candidates for anesthesia or surgery. METHODS AND RESULTS: Nine percutaneous fenestrations (one with additional stenting of the infrarenal true aortic lumen) were performed under local anesthesia in seven patients with aortic dissection. The presenting symptoms were abdominal angina or claudication. By the transfemoral approach, the intimal flap was initially punctured with a needle-catheter combination through which a guidewire was placed across the dissection flap. The fenestration was carried out with a balloon catheter introduced over the guidewire. The procedure was performed under on-line guidance with intravascular ultrasound imaging. The procedure was performed successfully and without complications in all patients. After intervention, symptoms resolved in all seven patients. CONCLUSIONS: Intravascular ultrasound-guided percutaneous fenestration of the intimal flap in symptomatic aortic dissections with distal vessel involvement is a technically feasible and safe procedure that can effectively relieve the patient's symptoms. PMID- 9337180 TI - The Munster Heart Study (PROCAM): total mortality in middle-aged men is increased at low total and LDL cholesterol concentrations in smokers but not in nonsmokers. AB - BACKGROUND: Some large epidemiological studies have shown an increase in mortality at low levels of total and LDL cholesterol. It has been speculated that low cholesterol levels may play a causative role in this association. To investigate this question, we analyzed all deaths occurring among middle-aged men in the Munster Heart Study (PROCAM), one of the largest prospective epidemiological studies of coronary heart disease risk markers in Europe. METHODS AND RESULTS: In the Munster Heart Study, 10,856 men aged 36 to 65 years at study entry (46.8+/-7.3 years [mean+/-SD]) were followed for 4 to 14 years (7.1+/-2.4 years). During this period, 313 deaths occurred--46 from myocardial infarction, 48 from suspected or definite sudden cardiac death, 14 from cerebrovascular disease, and 10 from other diseases of the circulatory system. There were 121 deaths from cancer and 33 deaths from violent causes (injuries in 16, suicide in 14, and homicide in 3 cases). Death in 29 cases occurred from other causes and was unexplained in 12 cases. Total cholesterol, LDL cholesterol, and the LDL/HDL ratio showed a J-shaped relationship with total mortality. At high total and LDL cholesterol concentrations, increased mortality was due to increased coronary deaths. At low total and LDL cholesterol concentrations, increased mortality was seen in smokers only and was explained by an increase in smoking-related cancer deaths. CONCLUSIONS: The increase in mortality at low levels of total and LDL cholesterol among middle-aged men in the Munster Heart Study is explained by an increase in smoking-related cancer deaths among smokers. PMID- 9337181 TI - Prevention of the angiographic progression of coronary and vein-graft atherosclerosis by gemfibrozil after coronary bypass surgery in men with low levels of HDL cholesterol. Lopid Coronary Angiography Trial (LOCAT) Study Group. AB - BACKGROUND: Studies have shown that treatment of hyperlipidemia, especially lowering of plasma LDL levels, retards the progression of coronary atherosclerosis and prevents clinical cardiovascular events. No such studies have focused on subjects with low levels of HDL cholesterol. METHODS AND RESULTS: We randomly assigned 395 post-coronary bypass men, who had an HDL cholesterol concentration < or = 1.1 mmol/L and LDL cholesterol < or = 4.5 mmol/L, to receive gemfibrozil 1200 mg/d or placebo. Coronary angiography was performed at baseline and after, on average, 32 months of therapy. Changes in coronary dimensions were assessed by computer-assisted analysis. Average on-trial serum triglyceride concentrations were 1.69+/-0.68 and 1.02+/-0.37, total cholesterol 5.48+/-0.68 and 4.83+/-0.63, LDL cholesterol 3.84+/-0.59 and 3.39+/-0.56, and HDL cholesterol 0.88+/-0.15 and 0.98+/-0.17 mmol/L in the placebo and gemfibrozil groups, respectively (mean+/-SD, each P<.001). The change in per-patient means of average diameters of native coronary segments was -0.04+/-0.11 mm in the placebo group and -0.01+/-0.10 mm in the gemfibrozil group (P=.009). The equivalent changes in minimum luminal diameters of stenoses were -0.09+/-0.18 and -0.04+/-0.15 mm, respectively (P=.002). A similar, albeit nonsignificant, trend toward treatment benefit was found in the predefined primary study end point, segments unaffected by grafts and those distal to graft insertions. In aortocoronary bypass grafts, 23 subjects (14%) assigned to placebo had new lesions in the follow-up angiogram, compared with 4 subjects (2%) assigned to gemfibrozil (P<.001). CONCLUSIONS: Gemfibrozil therapy retarded the progression of coronary atherosclerosis and the formation of bypass-graft lesions after coronary bypass surgery in men with low HDL cholesterol as their main lipid abnormality. PMID- 9337182 TI - Chlamydia pneumoniae, cytomegalovirus, and herpes simplex virus in atherosclerosis of the carotid artery. AB - BACKGROUND: Chlamydia pneumoniae and the herpes viruses cytomegalovirus (CMV) and herpes simplex virus type 1 (HSV-1) have been associated with human atherosclerosis in seroepidemiological and separate histopathological studies. We investigated the concurrent presence of these microorganisms in patients undergoing carotid endarterectomy. METHODS AND RESULTS: Endarterectomy specimens from 76 patients with carotid artery stenosis were stained for C. pneumoniae, CMV, and HSV-1 particles with specific IgG monoclonal antibodies by the avidin biotin-peroxidase method. IgG antibodies to CMV and C. pneumoniae were also measured in the serum. These were correlated with plaque morphology and the presence of the microorganisms in the atherosclerotic plaques. C. pneumoniae was detected in 54 (71%) (95% confidence interval [CI], 59.5% to 80.9%), CMV was detected in 27 (35.5%) (CI, 24.9% to 47.3%), and HSV-1 was detected in 8 (10.5%) (CI, 4.7% to 19.7%) versus none of 20 (0%) control normal carotid artery and aortic tissue (autopsy) specimens (CI, 0% to 16.8%) (P<.001 for CMV and C. pneumoniae). At least one microorganism was detected in 59 of the specimens (77.6%) (CI, 66.6% to 86.4%), with a single microorganism present only in 35 (46%), two microorganisms present in 18 (23.7%) (CI, 14.7% to 34.8%), and all three present in 6 (7.9%) (CI, 3.0% to 16.4%). Atherosclerotic plaques with thrombosis were more likely to have C. pneumoniae (80.4%) or CMV (57.8%) than were plaques without thrombosis (56.7% and 16.7%, respectively; P=.04 and .007). There was no correlation between the presence of CMV and C. pneumoniae in the atherosclerotic vessels and serum antibody titers. CONCLUSIONS: C. pneumoniae and CMV are commonly detected in atherosclerotic plaques of the carotid arteries, but their presence cannot be predicted by measuring serum antibodies. The presence of these microorganisms may predispose to a greater risk of thrombosis in the plaques, but further studies are needed to confirm this observation. PMID- 9337183 TI - Normalization of acquired QT prolongation in humans by intravenous potassium. AB - BACKGROUND: QT interval prolongation and dispersion have been implicated in serious arrhythmias in congestive heart failure (CHF) and the congenital and drug induced long-QT syndromes (LQTS). In a subset of the congenital LQTS, infusion of potassium can correct QT abnormalities, consistent with in vitro increases in outward currents such as I(Kr) or I(Kl) when extracellular potassium concentration ([K+]o) is increased. Furthermore, increasing [K+]o decreases the potency of I(Kr)-blocking drugs in vitro. The purpose of this study was to test the hypothesis that increasing [K+]o corrects QT abnormalities in CHF and in subjects treated with quinidine. METHODS AND RESULTS: KCl (maximum, 40 mEq) was infused into (1) 12 healthy subjects treated with quinidine sulfate (5 doses of 300 mg/5 h) or placebo and (2) 8 CHF patients and age-matched normal control subjects. Mean [K+] increased from 4 to 4.2 mEq/L to 4.7 to 5.2 mEq/L. Potassium infusion significantly reversed QTUc prolongation, especially in the precordial leads (quinidine, 590+/-79 to 479+/-35 [+/-SD] ms(1/2), P<.001; CHF, 521+/-110 to 431+/-47 ms(1/2), P<.05). There was no effect in either control group. Similarly, potassium decreased QTUc dispersion (quinidine, 210+/-62 to 130+/-75 ms(1/2), P<.01; CHF, 132+/-68 to 84+/-35 ms(1/2), P=.07) and was without effect in the control subjects. QT morphological abnormalities, including U waves and bifid T waves, were reversed by potassium. CONCLUSIONS: Potentially arrhythmogenic QT abnormalities during quinidine treatment and in CHF can be nearly normalized by modest elevation of serum potassium. PMID- 9337184 TI - Randomized, double-blind comparison of hirulog versus heparin in patients receiving streptokinase and aspirin for acute myocardial infarction (HERO). Hirulog Early Reperfusion/Occlusion (HERO) Trial Investigators. AB - BACKGROUND: Thrombolytic therapy improves survival after myocardial infarction through reperfusion of the infarct-related artery. Thrombin generated during thrombolytic administration may reduce the efficacy of thrombolysis. A direct thrombin inhibitor may improve early patency rates. METHODS AND RESULTS: Four hundred twelve patients presenting within 12 hours with ST-segment elevation were given aspirin and streptokinase and randomized in a double-blind manner to receive up to 60 hours of either heparin (5000 U bolus followed by 1000 to 1200 U/h), low-dose hirulog (0.125 mg/kg bolus followed by 0.25 mg x kg(-1) x h(-1) for 12 hours then 0.125 mg x kg(-1) x h(-1)), or high-dose hirulog (0.25 mg/kg bolus followed by 0.5 mg x kg(-1) x h(-1) for 12 hours then 0.25 mg x kg(-1) x h( 1)). The primary outcome was Thrombolysis In Myocardial Infarction trial (TIMI) grade 3 flow of the infarct-related artery at 90 to 120 minutes. TIMI 3 flow was 35% (95% CI, 28% to 44%) with heparin, 46% (95% CI, 38% to 55%) with low-dose hirulog, and 48% (95% CI, 40% to 57%) with high-dose hirulog (heparin versus hirulog, P=.023; heparin versus high-dose hirulog, P=.03). At 48 hours, reocclusion had occurred in 7% of heparin, 5% of low-dose hirulog, and 1% of high dose hirulog patients (P=NS). By 35 days, death, cardiogenic shock, or reinfarction had occurred in 25 heparin (17.9%), 19 low-dose hirulog (14%), and 17 high-dose hirulog patients (12.5%) (P=NS). Two strokes occurred with heparin, none with low-dose hirulog, and two with high-dose hirulog. Major bleeding (40% from the groin site) occurred in 28% of heparin, 14% of low-dose hirulog, and 19% of high-dose hirulog patients (heparin versus low-dose hirulog, P<.01). CONCLUSIONS: Hirulog was more effective than heparin in producing early patency in patients treated with aspirin and streptokinase without increasing the risk of major bleeding. Direct thrombin inhibition may improve clinical outcome. PMID- 9337186 TI - Phentolamine prevents adaptation to ischemia during coronary angioplasty: role of alpha-adrenergic receptors in ischemic preconditioning. AB - BACKGROUND: Experimental studies indicate that alpha-adrenergic receptors are involved in ischemic preconditioning. Their role in humans is unknown. METHODS AND RESULTS: Eighteen patients undergoing angioplasty for an isolated stenosis of the left anterior descending coronary artery were randomized to receive intravenous infusion of phentolamine or placebo during the procedure. Intracoronary ECG and cardiac pain were determined at the end of the first two balloon inflations. Average peak velocity in the contralateral coronary artery during balloon occlusion, an index of collateral recruitment, was also assessed by using a Doppler guide wire. In both phentolamine- and placebo-treated patients, average peak velocity significantly increased from baseline to the end of the first inflation (P<.01), but it did not show any further increase during the second inflation. In phentolamine-treated patients, ST-segment changes and cardiac pain severity during the second inflation were similar to those observed during the first inflation (13+/-9 versus 12+/-8 mm, P=NS, and 51+/-34 versus 54+/-32 mm, P=NS, respectively), whereas in placebo-treated patients, they were significantly less (6+/-4 versus 13+/-7 mm, P<.01, and 26+/-20 versus 49+/-22 mm, P<.05, respectively). CONCLUSIONS: The adaptation to ischemia observed in humans after two sequential coronary balloon inflations is abolished by phentolamine and is independent of collateral recruitment. Thus, it occurs due to ischemic preconditioning and is, at least in part, mediated by alpha-adrenergic receptors. PMID- 9337185 TI - Myocardial infarction and cardiac mortality in the Bypass Angioplasty Revascularization Investigation (BARI) randomized trial. AB - BACKGROUND: Cardiac mortality and myocardial infarction (MI) rates are used to evaluate the efficacy of coronary artery bypass grafting (CABG) and percutaneous transluminal coronary angioplasty (PTCA). We compared 5-year cardiac mortality and MI rates in 1829 patients with multivessel disease randomized to CABG or PTCA. METHODS AND RESULTS: The 5-year cardiac mortality rate was 8.0% in patients assigned to PTCA compared with 4.9% in those assigned to CABG (relative risk [RR] of 1.55 with a 95% confidence interval [CI] of 1.07 to 2.23; P=.022). In a subgroup of 1476 nondiabetic patients, there were no significant differences between treatment groups in cardiac mortality either overall (4.6% versus 4.2%; RR= 1.04, 95% CI, 0.65 to 1.66; P=.908) or in subgroups based on symptoms, left ventricular function, number of diseased vessels, or stenotic proximal left anterior descending artery. The two treatment groups had similar event rates for the combined end point of cardiac death or MI. The RR for cardiac mortality in 264 patients who sustained an MI compared with those who did not was 5.9 (P<.001). MIs were more common after CABG during index hospitalization (P=.004), but in the PTCA group, they were more common after discharge (P<.001). CONCLUSIONS: The Bypass Angioplasty Revascularization Investigation (BARI) trial indicates 5-year cardiac mortality in patients with multivessel disease was significantly greater after initial treatment with PTCA than with CABG. The difference was manifest in diabetic patients on drug therapy. There were no significant differences overall for the composite end point of cardiac mortality or MI between treatment groups or for cardiac mortality in nondiabetic patients regardless of symptoms, left ventricular function, number of diseased vessels, or stenotic proximal left anterior descending artery. PMID- 9337187 TI - Anti-ischemic and anti-anginal effects of thoracic epidural anesthesia versus those of conventional medical therapy in the treatment of severe refractory unstable angina pectoris. AB - BACKGROUND: Cardiac sympathetic blockade by thoracic epidural anesthesia (TEA) dilates stenotic coronary arteries and has been used to control pain in patients with unstable angina. The aim of the present study was to evaluate the potential anti-ischemic effects of cardiac sympathetic blockade by TEA in severe, refractory, unstable angina. METHODS AND RESULTS: Forty patients with unstable angina refractory to standard anti-anginal therapy were randomized to receive either continuous epidural infusion of bupivacaine (TEA, Th1 to Th5) or to standard anti-anginal therapy including beta-blockers, calcium antagonists, aspirin, heparin, and nitroglycerin infusion (control group). The primary end points were number of anginal attacks and severity of myocardial ischemia assessed by 48-hour ambulatory Holter monitoring. The incidence of myocardial ischemia was lower in the TEA group (22% versus 61%; P<.05). The number of ischemic episodes per patient was 1.0+/-0.6 in the TEA group and 3.6+/-0.9 in the control group (P<.05). The episode duration per patient was 4.1+/-2.5 minutes and 19.7+/-6.2 minutes in the TEA and the control groups, respectively (P<.05). The mean area-under-the-ST-time-curve was 6.8+/-4.3 and 32.2+/-14.3 (mm-min) in the TEA and the control groups, respectively (P<.05). Fifteen anginal attacks were recorded in the control group and one attack in the TEA group (0.83+/-0.21 versus 0.06+/-0.06/patient, respectively, P<.01). CONCLUSIONS: The anti-ischemic and anti-anginal effects of continuous TEA are superior to those of conventional therapy in the treatment of refractory unstable angina. PMID- 9337188 TI - Treatment of in-stent restenosis with excimer laser coronary angioplasty: mechanisms and results compared with PTCA alone. AB - BACKGROUND: This study determined the clinical safety, mechanisms, and 6-month results of excimer laser angioplasty (ELCA)+adjunct PTCA for the treatment of in stent restenosis and (via lesion matching) compared the results of ELCA+PTCA to PTCA alone. METHODS AND RESULTS: Using quantitative angiography (QCA) and intravascular ultrasound (IVUS), we studied 107 restenotic previously stented lesions in 98 patients before and after intervention. QCA measurements included minimum lumen diameter (MLD) and diameter stenosis (DS). IVUS measurements included stent, lumen, and intimal hyperplasia (IH=stent-lumen) cross-sectional areas (CSA) and volumes. In the 54 lesions treated with ELCA+PTCA, the MLD increased from 0.73+/-0.38 mm before ELCA to 2.10+/-0.47 mm after ELCA+PTCA (P<.0001); the DS decreased from 70+/-14% to 25+/-12% (P<.0001). By IVUS, the minimum lumen CSA increased from 1.58+/-0.78 mm2 before ELCA to 6.34+/-1.75 mm2 after ELCA+PTCA (P<.0001) as a result of an increase in minimum stent CSA from 7.70+/-2.41 to 9.10+/-2.60 mm2 (P<.0001) and a decrease in IH CSA from 5.25+/ 2.84 to 2.63+/-1.41 mm2 (P<.0001). Volumetric analysis showed that tissue ablation (during ELCA) contributed 29+/-15%, tissue extrusion (during adjunct PTCA) contributed 31+/-14%, and additional stent expansion (during adjunct PTCA) contributed 40+/-16% to the overall lumen gain. There were no ELCA-related complications. Matched to lesions treated with PTCA alone, ELCA+PTCA resulted in greater lumen gain, more IH ablation/extrusion, larger final lumen CSA (IVUS), and a tendency for less frequent need for subsequent target vessel revascularization (TVR, 21% versus 38%, P=.0823). CONCLUSIONS: ELCA safely and effectively ablates in-stent neointimal tissue. Adjunct PTCA extrudes neointimal tissue out of the stent and also further expands the stent. Compared with PTCA alone, ELCA+PTCA achieves better short-term and, potentially, better long-term results. PMID- 9337189 TI - Myocardial phosphocreatine-to-ATP ratio is a predictor of mortality in patients with dilated cardiomyopathy. AB - BACKGROUND: In patients with heart failure due to dilated cardiomyopathy, cardiac energy metabolism is impaired, as indicated by a reduction of the myocardial phosphocreatine-to-ATP ratio, measured noninvasively by 31P-MR spectroscopy. The purpose of this study was to test whether the phosphocreatine-to-ATP ratio also offers prognostic information in terms of mortality prediction as well as how this index compares with well-known mortality predictors such as left ventricular ejection fraction (LVEF) or New York Heart Association (NYHA) class. METHODS AND RESULTS: Thirty-nine patients with dilated cardiomyopathy were followed up for 928+/-85 days (2.5 years). At study entry, LVEF and NYHA class were determined, and the cardiac phosphocreatine-to-ATP ratio was measured by localized 31P-MR spectroscopy of the anterior myocardium. During the study period, total mortality was 26%. Patients were divided into two groups, one with a normal phosphocreatine to-ATP ratio (>1.60; mean+/-SE, 1.98+/-0.07; n=19; healthy volunteers: 1.94+/ 0.11, n=30) and one with a reduced phosphocreatine-to-ATP ratio (<1.60; 1.30+/ 0.05; n=20). At re-evaluation (mean, 2.5 years), 8 of 20 patients with reduced phosphocreatine-to-ATP ratios had died, all of cardiovascular causes (total and cardiovascular mortality, 40%). Of the 19 patients with normal phosphocreatine-to ATP ratios, 2 had died (total mortality, 11%), one of cardiovascular causes (cardiovascular mortality, 5%). Kaplan-Meier analysis showed significantly reduced total (P=.036) and cardiovascular (P=.016) mortality for patients with normal versus patients with low phosphocreatine-to-ATP ratios. A Cox model for multivariate analysis showed that the phosphocreatine-to-ATP ratio and NYHA class offered significant independent prognostic information on cardiovascular mortality. CONCLUSIONS: The myocardial phosphocreatine-to-ATP ratio, measured noninvasively with 31P-MR spectroscopy, is a predictor of both total and cardiovascular mortality in patients with dilated cardiomyopathy. PMID- 9337190 TI - Prognostic value of bisoprolol-induced hemodynamic effects in heart failure during the Cardiac Insufficiency BIsoprolol Study (CIBIS). AB - BACKGROUND: To further evaluate the mechanism of beta-blocker-induced benefits in heart failure, the relationships between bisoprolol-induced hemodynamic effects and survival were studied during the Cardiac Insufficiency BIsoprolol Study (CIBIS). METHODS AND RESULTS: In 557 patients studied, bisoprolol significantly reduced heart rate (-16.3+/-15.3 versus -1.6+/-13.4 bpm, respectively; P<.001) compared with placebo at 2 months after inclusion in the study. Heart rate change over time had the highest predictive value for survival (P<.01). Left ventricular fractional shortening (LVFS) significantly increased in the bisoprolol group compared with the placebo group 5 months after inclusion (+0.04+/-0.06 versus 0.001+/-0.05, respectively; P<.001; n=160). LVFS change over time was also significantly correlated with further survival (P<.02 by Cox analysis). Using a nonparametric approach, we demonstrated a significant interaction between study treatment group and LVFS over time. Patients who demonstrated improvement of LVFS over time (82% and 51% of patients in the bisoprolol and the placebo groups, respectively; P<.02) were at lower risk, but the hazard did not further decrease with a further increase of fractional shortening, and there was no significant difference between study treatment groups. Finally, it could be demonstrated that each of the three factors (heart rate change over time, LVFS change over time, and bisoprolol treatment) made a specific contribution to mortality rate. CONCLUSIONS: Preservation of left ventricular function appears to play a key role in the bisoprolol-induced beneficial effects on prognosis in heart failure. Short term beta-blocker-induced cardiac effects could provide a means to identify those patients who will experience improved survival over the long term. PMID- 9337191 TI - Pulmonary autograft procedure for aortic valve disease: long-term results of the pioneer series. AB - BACKGROUND: Pulmonary autograft replacement of the diseased aortic valve has not been widely practiced due to concerns regarding late autograft competence and the consequences of creating pulmonary valve disease. To investigate this, the fate of the pioneering series of patients has been determined. METHODS AND RESULTS: The 131 hospital survivors of the pulmonary autograft operation at the National Heart Hospital from 1967 to 1984 were identified and their outcomes determined to 1994. Age at operation was 11 to 52 years, and 109 patients were male. Autograft implantation was orthotopic subcoronary (107), free-standing root (20), or Dacron mounted (2). In 113 patients, homografts replaced the native pulmonary valve. Ten and 20 years after operation, survival was 85% and 61%, freedom from autograft replacement was 88% and 75%, and freedom from replacement of pulmonary position homografts was 89% and 80%, respectively. Causes of deaths (53) included chronic heart failure (13), complications of reoperation (12), and endocarditis (7). Autograft regurgitation, the most common indication for reoperation, appeared primarily technical in nature, usually due to cusp prolapse. Degeneration was found in only 3 of 30 explanted autografts, and the young patients showed no increase in late valve failure. Homografts outperformed other valve replacements in the pulmonary position, but patients with orthotopic subcoronary and root autografts survived similarly. CONCLUSIONS: The pulmonary autograft offers low rates of degeneration, endocarditis, and thromboembolism for a period lasting >20 years, particularly in the young, with reoperation mainly required for malpositioning of the autograft cusps. The capacity of the autograft to maintain viability with minimal degeneration is not matched by any other biological valve replacement. PMID- 9337192 TI - Paradoxical relationship between N-terminal proatrial natriuretic peptide and filling pressure in adults with cyanotic congenital heart disease. AB - BACKGROUND: Many adults with cyanotic congenital heart disease are characterized by reduced ventricular filling pressures and decreased systemic oxygen transport. Data from animals suggest that hypoxia can induce synthesis and secretion of atrial natriuretic peptide. METHODS AND RESULTS: We measured plasma N-terminal (1 98) proatrial natriuretic peptide (proANP) in 26 cyanotic adults and 28 noncyanotic control subjects. Resting arterial oxygen saturation was significantly lower and hemoglobin concentration and hematocrit significantly greater in cyanotic patients than in control subjects (82+/-6 versus 96+/-3%, 19.7+/-2.2 versus 14.7+/-2.1 g/dL, and 59.0+/-8.5% versus 44.3+/-5.2%, respectively, P<.0001 in all cases). Four cyanotic patients had evidence of iron deficiency. Plasma proANP levels were elevated in cyanotic patients compared with control subjects (1828+/-1147 versus 689+/-343 pmol/L, P<.0001). Comparison of resting arterial oxygen saturation and proANP levels demonstrated an inverse linear relationship between the two measures (r=-.70, P<.0001). There was a significant linear relationship between both hemoglobin concentration and hematocrit and proANP levels as well (r=.53, P=.0003 and r=.48, P=.002, respectively). Cyanotic patients had lower mean right atrial pressures than the control subjects (4+/-3 versus 7+/-2 mm Hg, P=.005), and there were inverse logarithmic relationships between proANP levels and systemic cardiac index (r= .82, P=.0002), systemic oxygen transport (r=-.68, P=.005), and mixed venous oxygen saturation (r=-.79, P<.0001). CONCLUSIONS: Adults with cyanotic congenital heart disease are characterized by increased levels of plasma proANP. The increased atrial natriuretic peptide most likely results in extracellular and plasma volume depletion and reduced systemic oxygen transport. Measures designed to increase ventricular filling may improve quality of life of these patients. PMID- 9337193 TI - Lung function and exercise gas exchange in chronic heart failure. AB - BACKGROUND: The ventilatory response to exercise in patients with chronic heart failure (HF) is greater than normal for a given metabolic rate. The objective of the present study was to determine the mechanism(s) for the high ventilatory output in patients with chronic HF. METHODS AND RESULTS: Centers in Germany, Italy, Japan, and the United States participated in this study. Each center contributed studies on patients and normal subjects of similar age and sex. One hundred thirty patients with chronic HF and 52 healthy subjects participated. Spirometric and breath-by-breath gas exchange measurements were made during rest and increasing cycle exercise. Arterial blood was sampled for measurement of pH, PaCO2, PaO2, and lactate during exercise in 85 patients. Resting forced expiratory volume in 1 second (FEV1) and vital capacity (VC) were proportionately reduced at all levels of impairment. Patients with more severe HF had greater tachypnea and a smaller tidal volume (VT) at a given exercise expired volume per unit time (VE). This was associated with an expiratory flow pattern characteristic of lung restriction. VE and VCO2 as a function of VO2 were increased during exercise in HF patients. The increases were greater the lower the peak VO2 per kilogram of body weight. The ratio of VD (physiological dead space) to VT and the difference between arterial and end tidal PCO2 at peak VO2 also increased inversely with peak VO2/kg. In contrast, the difference between alveolar and arterial PO2 and PaCO2 were both normal, on average, at peak VO2 regardless of the level of impairment. The more severe the exercise limitation, the higher the lactate and the lower the HCO3- at a given VO2, although pH was tightly regulated. CONCLUSIONS: The increase in VE in chronic HF patients is caused by an increase in VD/VT due to high ventilation/perfusion mismatching, an increase in VCO2 relative to VO2 resulting from HCO3- buffering of lactic acid, and a decrease in PaCO2 due to tight regulation of arterial pH. With regard to the excessive VE in HF patients, the increases in VD/VT and VCO2 relative to VO2 are more important as the patient becomes more exercise limited. Regional hypoperfusion but not hypoventilation typifies lung gas exchange in HF. This and other mechanisms might account for the restrictive changes leading to exercise tachypnea in HF patients. PMID- 9337194 TI - Size matters: the relationship between MMP-9 expression and aortic diameter. AB - BACKGROUND: Despite a wealth of data detailing increased metalloproteinase (MMP) 9 expression and activity in abdominal aortic aneurysms (AAAs), no studies examine the relationship between aortic size and MMP-9 expression. Because elastolysis occurs early in AAA formation, we hypothesized that MMP-9 expression would vary with aortic diameter. The purpose of this study was to measure MMP-9 mRNA levels in AAAs of various diameters and define the relationship between AAA size and MMP-9 expression. METHODS AND RESULTS: MMP-9 mRNA levels were measured by competitive polymerase chain reaction (PCR) using gene-specific external standards with cDNA from AAAs (n= 19) and normal aortas (n=4). Levels were normalized to GAPDH mRNA, determined separately via competitive PCR, to control for efficiency of reverse transcription. AAA size was measured on CT scans obtained within 6 weeks of surgery. MMP-9/GAPDH mRNA transcript levels in AAAs were expressed as mean+/-SEM and analyzed by ANOVA with a Tukey adjustment. There was a fourfold elevation in MMP-9/GAPDH mRNA transcript levels in 5.0- to 6.9-cm AAAs (98.06+/-15.19) compared with small (3.0- to 4.9-cm) AAAs (20.87+/-5.15, P<.03), large (>7-cm) AAAs (27.16+/-14.56, P<.01), or normal aortas (3.57+/-1.13, P<.003). The results did not change when they were normalized to patient height, nor were there significant differences in risk factors, age, or sex in each AAA group. CONCLUSIONS: MMP-9 mRNA expression is significantly higher in moderate diameter (5- to 6.9-cm) AAAs than either small (<4.0-cm) or large (>7.0-cm) AAAs. Increased MMP-9 expression may account for the propensity of AAAs >5 cm to continue to expand, in contrast to smaller aneurysms. Lower levels in AAAs >7 cm suggest that increases in other enzymes or in diameter-dependent mechanical stress on the aortic wall are responsible for their characteristic rapid expansion and high rupture rates. PMID- 9337195 TI - Flow-mediated dilation in 9- to 11-year-old children: the influence of intrauterine and childhood factors. AB - BACKGROUND: Early life factors, particularly size at birth, may influence later risk of cardiovascular disease, but a mechanism for this influence has not been established. We have examined the relation between birth weight and endothelial function (a key event in atherosclerosis) in a population-based study of children, taking into account classic cardiovascular risk factors in childhood. METHODS AND RESULTS: We studied 333 British children aged 9 to 11 years in whom information on birth weight, maternal factors, and risk factors (including blood pressure, lipid fractions, preload and postload glucose levels, smoking exposure, and socioeconomic status) was available. A noninvasive ultrasound technique was used to assess the ability of the brachial artery to dilate in response to increased blood flow (induced by forearm cuff occlusion and release), an endothelium-dependent response. Birth weight showed a significant, graded, positive association with flow-mediated dilation (0.027 mm/kg; 95% CI, 0.003 to 0.051 mm/kg; P=.02). Childhood cardiovascular risk factors (blood pressure, total and LDL cholesterol, and salivary cotinine level) showed no relation with flow mediated dilation, but HDL cholesterol level was inversely related (-0.067 mm/mmol; 95% CI, -0.021 to -0.113 mm/mmol; P=.005). The relation between birth weight and flow-mediated dilation was not affected by adjustment for childhood body build, parity, cardiovascular risk factors, social class, or ethnicity. CONCLUSIONS: Low birth weight is associated with impaired endothelial function in childhood, a key early event in atherogenesis. Growth in utero may be associated with long-term changes in vascular function that are manifest by the first decade of life and that may influence the long-term risk of cardiovascular disease. PMID- 9337196 TI - Pharmacological modulation of pressure-overload cardiac hypertrophy: changes in ventricular function, extracellular matrix, and gene expression. AB - BACKGROUND: Appropriate cardiac hypertrophy (CH) is necessary in several clinical settings, such as pulmonary artery banding in the two-stage arterial switch operation for transposition of the great arteries. Pressure-overload CH, however, produces ventricular dysfunction due to structural and molecular changes. The beta2-adrenergic receptor agonist clenbuterol has been shown to induce CH without such adverse effects to the rat heart. This study was performed to determine its effects on left ventricular (LV) function, structure, and gene expression in pressure-overload CH. METHODS AND RESULTS: Sprague-Dawley rats were assigned to one of four groups: 1, sham-operated (n=15); 2, banding of ascending aorta (n=22); 3, banding+clenbuterol (n=18); and 4, banding+thyroxine (n= 17). At the end of 3 weeks, groups 2, 3, and 4 showed an increase in LV mass index of 49.7+/ 5.1%, 66.1+/-3.8%, and 47.6+/-4.6%, respectively, relative to group 1. A subgroup with severe CH (>50%) in group 2 was found to have significantly impaired developed pressure and diastolic relaxation and an increase in passive stiffness, with significantly reduced LV expression of sarcoplasmic reticulum Ca2+-ATPase2a (SERCA2a) mRNA and increased LV collagen concentration. In comparison, similarly hypertrophied animals in groups 3 and 4 demonstrated improved developed pressure, normal relaxation and diastolic stiffness with normal collagen concentration, and a greater abundance of SERCA2a mRNA. CONCLUSIONS: Clenbuterol administration in conjunction with pressure overload produces a specific type of CH with preserved LV function. In addition, an increase in LV mass was associated with less fibrosis and greater expression of SERCA2a mRNA than banding alone. PMID- 9337197 TI - Regulation by differential development of Th1 and Th2 cells in peripheral tolerance to cardiac allograft induced by blocking ICAM-1/LFA-1 adhesion. AB - BACKGROUND: Specific immune tolerance to cardiac allografts is induced by anti ICAM-1 and anti-LFA-1 MAbs. Although the expression of the Th1 cytokines IL-2 and IFN-gamma is shown to increase in association with acute rejection, the roles of cytokines in the induction of peripheral tolerance by anti-ICAM-1 and anti-LFA-1 MAbs are not yet known. METHODS AND RESULTS: BALB/c hearts were transplanted into C3H/He mice. The MAbs to ICAM-1 and LFA-1 were injected for 3 days after transplantation in some recipients, and others were treated with FK506. IL-2 concentration in the supernatant of splenocytes from MAb-treated mice that were mix-cultured with donor splenocytes was lower than in normal controls. The expression of Th1 cytokines, detected by Northern blot assay, was enhanced in grafts or spleens of nontreated mice, whereas Th2 cytokines were expressed in the spleens of MAb-treated mice. No cytokine expression was enhanced in mice treated with FK506. Also, the induction of tolerance was prevented by the administration of rIL-2 in vivo in 5 of 7 mice, which were rendered tolerant. CONCLUSIONS: These data provide evidence that impairment of IL-2 production is critically involved in this tolerance induction and suggest that predominance of Th2 over Th1 cells is essential for tolerance induction by antiadhesion therapy. PMID- 9337199 TI - Advanced glycation end product (AGE)-mediated induction of tissue factor in cultured endothelial cells is dependent on RAGE. AB - BACKGROUND: Binding of advanced glycation end products (AGEs) to the cellular surface receptor (RAGE) induces translocation of the transcription factor NF kappaB into the nucleus and NF-kappaB-mediated gene expression. This study examines the role of RAGE in the AGE albumin-mediated induction of endothelial tissue factor, known to be partly controlled by NF-kappaB. METHODS AND RESULTS: Endothelial cells (ECs) were incubated in the presence of an 18-mer phosphorothioate oligodeoxynucleotide antisense to the 5'-coding sequence of the RAGE gene (antisense RAGE; 0.1 micromol/L). Sense oligonucleotides (sense RAGE, 0.1 micromol/L) of the same region served as control. The cellular uptake of oligonucleotides was controlled by immunofluorescence microscopy. RAGE transcription was suppressed by antisense RAGE, as demonstrated by RT-PCR reactions. AGE albumin-mediated activation of cultured ECs was studied after 48 hours of preincubation of ECs with antisense or sense RAGE. Electrophoretic mobility shift assays and Western blot analysis demonstrated that the AGE albumin induced translocation of NF-kappaB from the cytoplasm into the nucleus was suppressed in the presence of antisense RAGE but not by sense RAGE. In parallel, AGE albumin-mediated tissue factor transcription, activity, and antigen were significantly reduced in ECs exposed to antisense RAGE, whereas sense RAGE (and nonspecific oligonucleotides) did not influence tissue factor expression. CONCLUSIONS: Activation of ECs and induction of tissue factor by AGE albumin in ECs is dependent on RAGE. PMID- 9337198 TI - Adventitial gene transfer of recombinant endothelial nitric oxide synthase to rabbit carotid arteries alters vascular reactivity. AB - BACKGROUND: Adventitial gene transfer may serve as a tool to study vascular biology and may have therapeutic potential. We investigated the hypothesis that adenovirus-mediated transfer of the gene for endothelial nitric oxide synthase (eNOS) to the adventitia would alter vascular reactivity. METHODS AND RESULTS: Rabbit carotid arteries were surgically isolated and adenoviral vectors encoding eNOS (AdeNOS) or beta-galactosidase instilled into the periarterial sheath at a concentration of 1 x 10(10) pfu/mL. Arteries were harvested 4 days later for immunostaining, NOS enzymatic assay, measurement of cGMP, and vasomotor studies. Transgene expression in the adventitia was confirmed by histochemistry for beta galactosidase and immunostaining for eNOS with a monoclonal antibody. Calcium dependent NOS enzymatic activity and cGMP levels were significantly greater in the AdeNOS-transduced arteries. Maximal contractions to phenylephrine (10(-5) mol/L) were diminished in the AdeNOS-transduced arteries (4.6+/-0.2 versus 5.6+/ 0.2 g; P<.05), but in the presence of the eNOS inhibitor N(G)-monomethyl-L argininc (3x10(-4) mol/L) there was no difference between the two groups (7.1+/ 0.2 versus 7.5+/-0.3 g; P=NS). Relaxations to calcium ionophore obtained during submaximal contractions to phenylephrine were significantly enhanced in the AdeNOS-transduced arteries (-log EC50, 7.77+/-0.08 versus 7.45+/-0.07; P<.02). CONCLUSIONS: We conclude that eNOS gene transfer to the adventitia alters vascular reactivity, as demonstrated by diminished contractile responses to phenylephrine and enhanced relaxations to calcium ionophore. This may represent a therapeutic strategy for vascular diseases characterized by decreased bioavailability of NO. PMID- 9337200 TI - Local expression of C-type natriuretic peptide markedly suppresses neointimal formation in rat injured arteries through an autocrine/paracrine loop. AB - BACKGROUND: In vivo gene transfer into injured arteries may provide a new means to facilitate molecular understanding of and to treat the intractable fibroproliferative arterial diseases. Selection of an optimal molecule to be transferred will be a key to successful gene therapy in the future. We tested the hypothesis that a secreted multifactorial molecule should act more efficiently through an autocrine/paracrine loop to suppress neointimal formation elicited in injured arteries than a simple growth-inhibiting molecule that might be expressed inside cells. METHODS AND RESULTS: We constructed an adenoviral vector (AdCACNP) expressing C-type natriuretic peptide (CNP), a secreted stimulator of membrane bound guanyl cyclase. AdCACNP directs cells to secrete large quantities of biologically active CNP. Serum-stimulated DNA synthesis and cell proliferation were only moderately suppressed in arterial smooth muscle cells infected with AdCACNP in vitro. However, when AdCACNP was applied to balloon-injured rat carotid arteries in vivo, neointimal formation was markedly reduced (90% reduction) in an infection-site-specific manner without an increase in plasma CNP level. CONCLUSIONS: Our results showed that CNP, a secreted multifactorial molecule, was indeed effective in suppressing fibroproliferative response in injured arteries and suggest that the potent antiproliferation effect may not be the most critical factor for the effective suppression of neointimal formation. An adenovirus-mediated expression of CNP could be an effective and site-specific form of molecular intervention in proliferative arterial diseases. PMID- 9337201 TI - Effect of serotonin, thromboxane A2, and specific receptor antagonists on vascular smooth muscle cell proliferation. AB - BACKGROUND: Restenosis is a major complication that limits the long-term efficacy of coronary angioplasty. Migration and proliferation of activated medial smooth muscle cells (SMCs) is considered an important mechanism in this process. Because at sites of vascular injury, aggregating platelets release both serotonin (5-HT) and thromboxane A2 (TXA2), we examined whether 5-HT and TXA2 can induce SMC proliferation and whether there is synergistic interaction between these two mediators. METHODS AND RESULTS: The mitogenic effects of 5-HT and TXA2 either alone or in combination was examined in serum-free medium on canine aortic SMCs by [3H]thymidine incorporation into DNA and by cell counting. 5-HT induced SMC proliferation at a concentration of 100 nmol/L, whereas the effect of TXA2 (U46619, a stable TXA2 mimetic) on inducing proliferation of SMCs was observed at a concentration of 100 nmol/L. When these two mediators were added together, there was a synergistic interaction on inducing SMC proliferation even at subthreshold concentrations. The mitogenic effect of 5-HT and its synergistic interaction with TXA2 on SMC proliferation was abolished by a 5-HT2 receptor antagonist, LY281067, without affecting the contribution of TXA2. Similarly, the TXA2 synthase inhibitor/receptor antagonist ridogrel abolished the mitogenic effect of TXA2 and the interaction between 5-HT and TXA2 without affecting the response to 5-HT. When LY281067 and ridogrel were used together, they abolished the mitogenic effects of 5-HT and TXA2. CONCLUSIONS: At sites of vascular injury, platelet-induced SMC proliferation may also be modulated by nonpeptide growth mediators. A combination of a 5-HT2 receptor antagonist and TXA2 synthase inhibitor/receptor may be useful for attenuation of restenosis after angioplasty. PMID- 9337202 TI - Simultaneous overexpression of two stress proteins in rat cardiomyocytes and myogenic cells confers protection against ischemia-induced injury. AB - BACKGROUND: Mitochondria are known to be a major target during ischemic cardiac injury. Previous studies have shown that in rodent myogenic cells and in the hearts of transgenic mice in which the heat shock or stress protein 70 is increased, there is a marked tolerance to ischemia/reperfusion injury. Two other heat shock proteins (HSP60 and HSP10) are known to form, within the mitochondria, a chaperonin complex that is important for mitochondrial protein folding and function. We were then interested in investigating whether increased expression of these two stress proteins is able to protect myogenic cells against ischemia/reperfusion injury. METHODS AND RESULTS: We generated recombinant adenoviral vectors containing HSP60, HSP10, or a combination of the two genes. These adenoviral constructs overexpress significant amounts of these stress proteins in both rat neonatal cardiomyocytes and the myogenic H9 c2 cell line. Cells infected with an adenoviral construct overexpressing both HSP60 and HSP10 were found to be protected against simulated ischemia, whereas cells infected with adenoviral constructs overexpressing only HSP60 or HSP10 alone were not rendered tolerant to simulated ischemic injury. CONCLUSIONS: These results suggest that the simultaneous expression of these two proteins that form a chaperonin complex in the mitochondria plays an important role in the survival of myogenic cells after ischemia/reperfusion injury. PMID- 9337203 TI - Local delivery of ethanol inhibits intimal hyperplasia in pig coronary arteries after balloon injury. AB - BACKGROUND: Smooth muscle cell (SMC) hyperplasia is an important mechanism of restenosis after coronary angioplasty and the primary mechanism of restenosis within coronary stents. Ethanol has been shown to reduce the response of SMCs to local growth stimulants in vitro. This study was carried out to determine whether local delivery of ethanol solution could reduce intimal hyperplasia induced by balloon injury. METHODS AND RESULTS: Three groups of juvenile domestic pigs underwent oversized balloon dilation injury of the left anterior descending and left circumflex coronary arteries. Immediately after the balloon injury, one of the arteries was randomized to local delivery of 15% ethanol with a local delivery balloon catheter, and the other received no further treatment. Histological and morphometric studies were carried out at 2 weeks in group 1 (n=16) and at 4 weeks in group 2 (n=10). In the third group (n=15), animals were killed at days 4, 8, and 14 after balloon injury, and coronary artery segments were studied by immunohistochemical staining against proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU). Histological injury scores were not different between the ethanol-treated and untreated arterial segments in either group 1 or 2. The neointimal areas were significantly smaller in the ethanol treated arterial segments than in the untreated segments (0.25+/-0.08 versus 0.57+/-0.08 mm2, P=.004, at 2 weeks; 0.33+/-0.05 versus 0.54+/-0.07 mm2, P=.03, at 4 weeks). SMC proliferative activity was significantly lower in ethanol treated arteries than in untreated arteries at 4 and 8 days after injury by BrdU and PCNA staining. CONCLUSIONS: Local delivery of 15% ethanol solution to pig coronary arteries significantly decreased the SMC proliferative activity and neointimal formation induced by balloon dilation injury. PMID- 9337204 TI - Antithrombin III prevents and rapidly reverses leukocyte recruitment in ischemia/reperfusion. AB - BACKGROUND: P-selectin has recently been shown to be essential for leukocyte rolling after the reperfusion of ischemic mesentery. However, the mediators responsible for neutrophil rolling in ischemic microvessels remain entirely unclear. METHODS AND RESULTS: Intravital microscopy was used to examine leukocyte kinetics in a feline mesentery ischemia/reperfusion model. Sixty minutes of ischemia followed by reperfusion caused a profound increase in leukocyte rolling and adhesion. Pretreatment with the endogenous antithrombotic agent antithrombin III (ATIII) infused as a bolus (250 U/kg) reduced neutrophil rolling and adhesion to preischemic levels during reperfusion. No effect was seen with heat-inactive ATIII. Importantly, ATIII posttreatment also significantly reduced neutrophil rolling and adhesion during reperfusion, suggesting that ATIII can reverse the leukocyte recruitment response induced by ischemia/reperfusion. Vascular permeability was also reduced by 50% after ATIII administration. To determine whether ATIII could reverse thrombin-induced rolling directly, neutrophil rolling was performed on human endothelium in flow chambers. Indeed, thrombin-induced rolling, but not histamine-induced rolling, could be rapidly reversed with ATIII on endothelium, suggesting that ATIII affects thrombin rather than directly affecting neutrophils or the endothelium. CONCLUSIONS: This study demonstrates for the first time that thrombin plays an important role in ischemia-induced leukocyte rolling and adhesion and that ATIII can be used therapeutically postreperfusion to attenuate the leukocyte recruitment response in inflammation without the nonspecific effects associated with anti-adhesion molecule therapy. PMID- 9337205 TI - Delayed coronary endothelial protection 24 hours after preconditioning: role of free radicals. AB - BACKGROUND: Preconditioning (PC) induces delayed protection against myocardial ischemia/reperfusion (I/R) injury. Whether a similar late protective effect exists in coronary endothelial cells is not known. Thus, we assessed whether PC also induces late protection against endothelial injury after I/R and the potential role of reactive oxygen species as triggers for late PC in this setting. METHODS AND RESULTS: Rats were subjected to sham surgery or to PC with 1 cycle of 2 minutes I/5 minutes R and 2 cycles of 5 minutes I/5 minutes R in the absence or presence of the free radical scavenger N-2-mercaptopropionyl glycine (MPG). Twenty-four hours later, rats were subjected to 20 minutes I/60 minutes R in the absence or presence of MPG. At the end of R, coronary segments (diameter, 200 to 300 microm) were removed distal to the site of occlusion and mounted in wire myographs. I/R reduced the relaxations to acetylcholine (maximal relaxations: sham, 72+/-6%; I/R, 31+/-6%), and this impairment was prevented by MPG (64+/-7%). PC improved the response to acetylcholine (48+/-6%), but this beneficial effect was abolished by MPG (23+/-5%). CONCLUSIONS: PC induces late protection against reperfusion-induced coronary endothelial injury. Moreover, in addition to being mediators of endothelial injury during reperfusion after prolonged ischemia, reactive oxygen species produced during PC also protect the coronary endothelium from reperfusion injury 24 hours later. PMID- 9337207 TI - Myocardial uptake and redistribution of 99mTc-N-NOET in dogs with either sustained coronary low flow or transient coronary occlusion: comparison with 201Tl and myocardial blood flow. AB - BACKGROUND: 99mTc-N-NOET (NOET) is a new myocardial perfusion imaging agent that redistributes over time. We sought to better define the redistribution kinetics of NOET using open-chest canine models of sustained low coronary flow (protocol 1) and transient coronary occlusion followed by reflow (protocol 2). METHODS AND RESULTS: In protocol 1 (n=10), NOET and 201Tl were injected during low flow in the left anterior descending coronary artery (LAD) that was sustained for 2 hours. Protocol 2 dogs (n=6) were injected with NOET during 20 minutes of LAD occlusion followed by 2 hours of reflow. In both protocols, serial NOET planar images were acquired, and myocardial flow and 2-hour tracer activities were determined by gamma-well counting. Defect resolution was observed on images in both protocols. Initial defect count ratios, reflecting flow disparity at injection (0.66+/-0.03 and 0.57+/-0.04, respectively), increased over 2 hours (0.73+/-0.02 and 0.75+/-0.04, respectively; P<.001 versus initial). Quantitative imaging showed that NOET redistribution resulted from greater clearance from normal areas versus low-flow or transiently occluded areas. In protocol 1, 2-hour NOET and 201Tl stenotic-to-normal tissue activity ratios were similar (0.76+/ 0.06 versus 0.73+/-0.04, P=NS) and higher than injection flow ratios (0.52+/-0.06 and 0.56+/-0.07, respectively, P<.001), consistent with tracer redistribution. In protocol 2, NOET redistributed to an even greater extent (injection flow ratio, 0.27+/-0.04; 2-hour tissue activity ratio, 0.84+/-0.03, P<.001). CONCLUSIONS: NOET is the first 99mTc-labeled myocardial imaging agent with kinetics similar to 201Tl in experimental models, permitting redistribution imaging. NOET appears to be a promising agent for assessing patients with coronary artery disease. PMID- 9337206 TI - Peroxynitrite reduces myocardial infarct size and preserves coronary endothelium after ischemia and reperfusion in cats. AB - BACKGROUND: Peroxynitrite (ONOO-) is purported to exert cytotoxic effects at high doses. However, physiologically relevant concentrations of ONOO- inhibit polymorphonuclear neutrophil (PMN) adhesion to the endothelium and attenuate PMN mediated contractile dysfunction in isolated perfused rat hearts. We are unaware of any reports in vivo showing effects of peroxynitrite in myocardial ischemia and reperfusion (MI/R). Thus, the purpose of this study was to examine the in vivo effects of a physiologically relevant concentration of ONOO- (1 micromol/L) in a feline model of MI/R injury. METHODS AND RESULTS: ONOO- (1 micromol/L) or its vehicle (0.9% NaCl at pH 8.4) was infused intraventricularly, starting 10 minutes before reperfusion in cats subjected to 90 minutes of myocardial ischemia and 4.5 hours of reperfusion. ONOO(-)-treated cats demonstrated marked attenuation of cardiac necrosis after MI/R compared with cats receiving only vehicle (P<.001). Moreover, vasorelaxation of ischemic-reperfused left anterior descending (LAD) coronary artery rings in response to the endothelium-dependent dilators acetylcholine and A23187 was greater in rings isolated from ONOO(-) treated MI/R cats compared with MI/R cats receiving only vehicle, indicating that postreperfusion coronary vascular endothelial function was preserved by ONOO-. ONOO- also significantly reduced adherence of neutrophils to the ischemic reperfused LAD coronary endothelium. Immunohistochemical localization of P selectin was also significantly attenuated in hearts from ONOO(-)-infused MI/R cats. CONCLUSIONS: These data suggest that physiologically relevant concentrations of ONOO- exert significant cardioprotective and vasculoprotective effects in MI/R in cats, at least partially by attenuating PMN-endothelium interactions. PMID- 9337208 TI - Myocardial 99mTc-tetrofosmin uptake during adenosine-induced vasodilatation with either a critical or mild coronary stenosis: comparison with 201Tl and regional myocardial blood flow. AB - BACKGROUND: Clinical studies have shown a comparable coronary stenosis detection rate between 99mTc-tetrofosmin and 201Tl but with smaller defect magnitudes for 99mTc-tetrofosmin. The goals of this study were to measure the first-pass extraction fraction (EF) of 99mTc-tetrofosmin in canine myocardium and to compare 99mTc-tetrofosmin with 201Tl uptake during adenosine-induced vasodilatation in dogs with various degrees of coronary stenosis. METHODS AND RESULTS: EF was calculated in 4 anesthetized, open-chest dogs after intracoronary administration of 125I-labeled albumin and 99mTc-tetrofosmin. In another 16 dogs with either critical (n=6) or mild (n= 10) left anterior descending coronary artery (LAD) stenoses, 201Tl and 99mTc-tetrofosmin were administered during adenosine infusion (250 microg x kg(-1) x min(-1)). Dogs were killed 5 minutes later, and tracer activities were determined by ex vivo imaging of heart slices and by well counting. Mean 99mTc-tetrofosmin EF was 54.0+/-3.7%. In the 6 critical-stenosis dogs, the LAD-to-left circumflex artery (LCx) microsphere flow ratio was 0.22+/ 0.02 with adenosine. The LAD-to-LCx activity ratios were 0.37+/-0.04 for 201Tl and 0.67+/-0.05 for 99mTc-tetrofosmin (P<.01). For the 10 mild-stenosis dogs, the LAD-to-LCx flow ratio was 0.44+/-0.05. The 201Tl activity ratio was 0.58+/-0.04, compared with 0.81+/-0.04 for 99mTc-tetrofosmin (P<.01). Thus, in both groups, 99mTc-tetrofosmin uptake underestimated the flow disparity more than 201Tl. Similarly, magnitudes of ex vivo image defects were significantly greater for 201Tl than for 99mTc-tetrofosmin in both groups. CONCLUSIONS: In this canine model, relative underperfusion with adenosine stress is better resolved with 201Tl than with 99mTc-tetrofosmin and may be explained by the lower EF for 99mTc tetrofosmin. With clinical imaging, greater 201Tl attenuation and redistribution may lessen this advantage. PMID- 9337209 TI - Inhibition of arterial thrombosis by recombinant annexin V in a rabbit carotid artery injury model. AB - BACKGROUND: The procoagulant effect of anionic phospholipid may play a major role in the development of arterial thrombosis. METHODS AND RESULTS: Annexin V, a calcium-dependent anionic-phospholipid-binding protein, was expressed and isolated from Escherichia coli and its antithrombotic effect examined in a rabbit carotid artery thrombosis model. A partially occlusive thrombus was formed in the left carotid artery by application of electric current to produce an approximately 50% occlusion of the lumen. After the current was discontinued, flow ceased completely within 42+/-12 minutes (n=6) because of continuing platelet/fibrin thrombus formation. When annexin V was given at doses of 2.8 to 16.6 microg x kg(-1) x min(-1) for a period of 180 minutes, starting at the time the current was stopped, there was a dose-dependent inhibition of thrombus formation. At a dose of 5.6 microg x kg(-1) x min(-1), blood flow remained patent throughout the infusion and for an additional 60 minutes after the infusion was stopped. In addition, there was a decrease in thrombus weight (16+/-7.4 versus 2.0+/-1.0 g), (125)I-fibrin deposition (approximately 45% reduction, P<.001), and (111)In-labeled platelet accumulation (approximately 43% reduction, P<.001). Prior mixing of annexin V with phosphatidylserine micelles abolished the antithrombotic effect of annexin V, whereas mixing with phosphatidylcholine micelles had no effect. The antithrombotic effect of annexin V was not associated with bleeding tendency, as judged by the amount of blood absorbed in a gauze pad placed in a surgical incision extending to the muscle tissue and by the standard template bleeding time. CONCLUSIONS: These observations support a potentially important role for anionic phospholipid exposure in platelets in arterial thrombosis, and inhibition of this activity could be a novel target for therapy in coronary thrombosis and stroke and after angioplasty. PMID- 9337210 TI - Decrease in forces responsible for diastolic suction during acute coronary occlusion. AB - BACKGROUND: The production of left ventricular (LV) restoring forces generated during contraction, which are responsible for diastolic suction, is dependent on end-systolic volume (ESV) and systolic transmural and 3D deformation. We tested the hypothesis that acute coronary occlusion would result in loss of forces that cause suction. METHODS AND RESULTS: Ten open-chest dogs were subjected to a 10 minute acute coronary occlusion (proximal left anterior descending coronary artery). A servomotor connected to the left atrium (LA) was used to rapidly clamp LA pressure during systole below the level of the succeeding LV diastolic pressure, resulting in nonfilling diastoles during which the LV fully relaxed at its ESV. LA clamps at multiple ESVs (conductance catheter) allowed delineation of positive and negative portions of the fully relaxed LV pressure-volume relation (FRPVR). A negative fully relaxed pressure (FRP) indicates the presence of restoring forces. After 10 minutes of acute coronary occlusion, there was an upward shift of the FRPVR. Thus, for example, at matched ESVs before and during coronary occlusion, FRP was -1.1+/-1.1 (+/-SD) mm Hg before versus 0.2+/-1.2 mm Hg after 10 minutes of coronary occlusion (P<.05). CONCLUSIONS: Acute coronary occlusion results in a rapid decrease in forces responsible for suction. This phenomenon is independent of the level of ESV and may contribute to ischemic diastolic dysfunction. PMID- 9337211 TI - Effects of dobutamine stress on myocardial blood flow, 99mTc sestamibi uptake, and systolic wall thickening in the presence of coronary artery stenoses: implications for dobutamine stress testing. AB - BACKGROUND: Although dobutamine stress is used with both 99mTc sestamibi (sestamibi) myocardial perfusion imaging and echocardiography for detecting coronary artery stenoses, the impact of stenosis severity on test end points (myocardial sestamibi uptake and systolic thickening, respectively) has not been clearly defined. METHODS AND RESULTS: In 15 open-chest dogs, dobutamine (2.5 to 30 microg x kg(-1) x min(-1)) was infused after placement of an LAD stenosis that reduced (n=8) or abolished (n=7) flow reserve. In dogs with reduced flow reserve, the stenotic-to-normal sestamibi activity ratio (0.86+/-0.03) significantly underestimated the approximately 2-to-1 dobutamine-induced flow disparity at the time of sestamibi injection (flow ratio, 0.53+/-0.04; P<.001). Stenotic-zone thickening increased at low but not at higher doses of dobutamine (2.9+/-0.4 versus 4.2+/-0.4 mm in normal zone at peak dobutamine; P=.055) but did not fall below baseline (2.7+/-0.3 mm). Similarly, in dogs with absent flow reserve, the sestamibi activity ratio (0.78+/-0.02) underestimated the approximately 2.5-to-1 dobutamine-induced flow disparity (flow ratio, 0.41+/-0.05; P<.001), and failure to increase systolic thickening was observed in the stenotic zone (2.7+/-0.4 versus 4.6+/-0.3 mm in the normal zone at peak stress, P<.01). In both groups of dogs, myocardial sestamibi uptake and image defect magnitudes were less than expected for the dobutamine-induced hyperemia, suggesting that dobutamine adversely affects myocardial sestamibi binding. Finally, a significant reduction in stenotic-zone thickening was seen during postdobutamine recovery, consistent with myocardial stunning. CONCLUSIONS: In the presence of stenoses that reduced or abolished regional flow reserve, (1) myocardial sestamibi uptake significantly underestimated the dobutamine-induced flow heterogeneity, (2) a "failure to increase systolic thickening" rather than a reduction in thickening was observed during dobutamine stress, and (3) myocardial stunning was observed during postdobutamine recovery. PMID- 9337213 TI - Relation between muscle contraction speed and hydraulic performance in skeletal muscle ventricles. AB - BACKGROUND: The fatigue resistance and power-to-weight ratio of skeletal muscle that has been conditioned by electrical stimulation makes cardiac assistance from a graft of such muscle a realistic prospect. A skeletal muscle must be surgically reconfigured to act on the circulating blood, but little is known about the power losses that accompany such interventions. We investigated in acute experiments the hydraulic performance of approximately cylindrical pumps made from sheep latissimus dorsi (LD) muscles, having first characterized the performance of each muscle in situ. METHODS AND RESULTS: Force-length and force-velocity relations were measured in situ for LD that had received either 8 weeks of stimulation at 2 Hz or no chronic stimulation. Two sizes of skeletal muscle ventricle (SMV) were formed from the same muscles, and their hydraulic performance was measured. The hydraulic performance was also calculated from the linear data, models of the force-length and force-velocity curves, and a description of the stress distribution within the SMV wall. The model predicted well the isovolumetric function of the ventricles and the optimum afterload but overestimated the flow and therefore the power. In conditioned ventricles the performance was particularly poor because of the slow contractile properties of the muscles. CONCLUSIONS: If SMVs are to pump effectively against the arterial impedance, the pressure drop caused by flow (or the internal resistance) should be lower than that of the ventricles we constructed. Progress can be made through refinement of surgical technique and stimulation protocols that generate faster fatigue resistant muscles. PMID- 9337212 TI - Platelet-derived growth factor-stimulated superoxide anion production modulates activation of transcription factor NF-kappaB and expression of monocyte chemoattractant protein 1 in human aortic smooth muscle cells. AB - BACKGROUND: Platelet-derived growth factor (PDGF) and superoxide anion (O2.-) have been implicated in vascular diseases. We investigated whether PDGF stimulates the production of O2.- in human aortic smooth muscle cells (HSMCs) and whether O2.- leads in this way to the activation of nuclear factor-kappaB (NF kappaB) and induction of monocyte chemoattractant protein 1 (MCP-1) in PDGF stimulated HSMCs. METHODS AND RESULTS: PDGF-AB concentration- and time dependently stimulated O2.- generation from HSMCs. The stimulatory effect of PDGF AB was mimicked by PDGF-BB but not by PDGF-AA. The generation of O2.- by PDGF-AB was attenuated by the NAD(P)H oxidase inhibitor iodonium diphenyl, the specific protein kinase C (PKC) inhibitor Ro 31-8220, and the phosphatidylinositol 3 kinase inhibitor wortmannin. Allopurinol and nifedipine had no effect on PDGF-AB induced O2.- release, whereas indomethacin potentiated this response. Gel mobility shift assay revealed that PDGF-AB increased the binding activity of NF kappaB, which contained predominantly the p50/p65 heterodimer in nuclear extracts from HSMCs. Superoxide dismutase as well as iodonium diphenyl, Ro 31-8220, and wortmannin attenuated PDGF-AB-induced activation of NF-kappaB and expression of MCP-1 mRNA. In contrast, superoxide dismutase did not inhibit the interleukin 1beta-induced NF-kappaB activation. CONCLUSIONS: The results demonstrate that PDGF stimulates O2.- generation in HSMCs via PKC-dependent and wortmannin sensitive pathways involving flavoenzyme(s). This PDGF-induced O2.- production may be involved in vascular lesion formation by mediating, at least in part, NF kappaB activation and MCP-1 induction. PMID- 9337214 TI - Preservation of myocyte contractile function after hyperthermic cardioplegic arrest by activation of ATP-sensitive potassium channels. AB - BACKGROUND: Left ventricular (LV) dysfunction can occur after hyperkalemic cardioplegic arrest and subsequent reperfusion and rewarming. Activation of adenosine triphosphate (ATP)-sensitive potassium (KATP) channels within the myocyte sarcolemma has been shown to be cardioprotective for myocardial reperfusion injury and ischemia and may play a contributory role in preconditioning for cardioplegic arrest. Accordingly, the present study tested the hypothesis that cardioplegic arrest and activation of KATP channels by a potassium channel opener (PCO) would attenuate alterations in ionic homeostasis and improve myocyte contractile function. METHODS AND RESULTS: Porcine LV myocytes were isolated and randomly assigned to the following treatment groups: normothermic control, incubation in cell culture media for 2 hours at 37 degrees C (n=60); hyperkalemic cardioplegia, incubation for 2 hours in hypothermic hyperkalemic cardioplegic solution (n=60); or PCO/cardioplegia, incubation in cardioplegic solution containing 100 micromol/L of the PCO aprikalim (n=60). Hyperkalemic cardioplegia and rewarming caused a significant reduction in myocyte velocity of shortening compared with normothermic control values (33+/-2 versus 66+/-2 microm/s, P<.05). Cardioplegic arrest with PCO supplementation significantly improved indices of myocyte contractile function when compared with hyperkalemic cardioplegia (58+/-4 microm/s, P<.05). Myocyte intracellular calcium increased during hyperkalemic cardioplegic arrest compared with baseline values (147+/-2 versus 85+/-2 nmol/L, P<.05). The increase in intracellular calcium was significantly reduced in myocytes exposed to the PCO-supplemented cardioplegic solution (109+/-4 nmol/L, P<.05). CONCLUSIONS: Cardioplegic arrest with simultaneous activation of KATP channels preserves myocyte contractile processes and attenuates the accumulation of intracellular calcium. These findings suggest that changes in intracellular calcium play a role in myocyte contractile dysfunction associated with cardioplegic arrest. Moreover, alternative strategies may exist for preservation of myocyte contractile function during cardioplegic arrest. PMID- 9337215 TI - Modulation of the renin-angiotensin pathway through enzyme inhibition and specific receptor blockade in pacing-induced heart failure: I. Effects on left ventricular performance and neurohormonal systems. AB - BACKGROUND: The goal of this study was to determine the effects of ACE inhibition (ACEI) alone, AT1 angiotensin (Ang) II receptor blockade alone, and combined ACEI and AT1 Ang II receptor blockade on LV function, systemic hemodynamics, and neurohormonal system activity in a model of congestive heart failure (CHF). METHODS AND RESULTS: Pigs were randomly assigned to each of 5 groups: (1) rapid atrial pacing (240 bpm) for 3 weeks (n=9), (2) ACEI (benazeprilat, 0.187 mg x kg( 1) x d(-1)) and rapid pacing (n=9), (3) AT1 Ang II receptor blockade (valsartan, 3 mg x kg(-1) x d(-1)) and rapid pacing (n=9), (4) ACEI and AT1 Ang II receptor blockade (benazeprilat/valsartan, 0.05/3 mg x kg(-1) d(-1)) and rapid pacing (n=9), and (5) sham controls (n=10). In the pacing group, LV fractional shortening (LVFS) fell (13.4+/-1.4% versus 39.1+/-1.0%) and end-diastolic dimension (LVEDD) increased (5.61+/-0.11 versus 3.45+/-0.07 cm) compared with control (P<.05). With AT1 Ang II blockade and rapid pacing, LVEDD and LVFS were unchanged from pacing-only values. ACEI reduced LVEDD (4.95+/-0.11 cm) and increased LVFS (20.9+/-1.9%) from pacing-only values (P<.05). ACEI and AT1 Ang II blockade reduced LVEDD (4.68+/-0.07 cm) and increased LVFS (25.2+/-0.9%) from pacing only (P<.05). Plasma norepinephrine and endothelin increased by more than fivefold with chronic pacing and remained elevated with AT1 Ang II blockade. Plasma norepinephrine was reduced from pacing-only values by more than twofold in the ACEI group and the combination group. ACEI and AT1 Ang II receptor blockade reduced plasma endothelin levels by >50% from rapid-pacing values. CONCLUSIONS: These findings suggest that the effects of ACEI in the setting of CHF are not solely due to modulation of Ang II levels but rather to alternative enzymatic pathways and that combined ACEI and AT1 Ang II receptor blockade may provide unique benefits for LV pump function and neurohormonal systems in the setting of CHF. PMID- 9337216 TI - Modulation of the renin-angiotensin pathway through enzyme inhibition and specific receptor blockade in pacing-induced heart failure: II. Effects on myocyte contractile processes. AB - BACKGROUND: The goal of this study was to determine the effects of ACE inhibition alone, AT1 angiotensin (Ang) II receptor blockade alone, and combined ACEI and AT1 Ang II receptor blockade in a model of congestive heart failure (CHF) on isolated LV myocyte function and fundamental components of the excitation contraction coupling process. METHODS AND RESULTS: Pigs were randomly assigned to one of five groups: (1) rapid atrial pacing (240 bpm) for 3 weeks (n=9), (2) concomitant ACEI (benazeprilat, 0.187 mg x kg(-1) x d(-1)) and rapid pacing (n=9), (3) concomitant AT1 Ang II receptor blockade (valsartan, 3 mg/kg/d) and rapid pacing (n=9), (4) concomitant ACEI and AT1 Ang II receptor blockade (benazeprilat/valsartan, 0.05/3 mg x kg(-1) x d(-1)) and rapid pacing (n=9), and (5) sham controls (n=10). LV myocyte shortening velocity was reduced with chronic rapid pacing compared with control (27.2+/-0.6 versus 58.6+/-1.2 microm/s, P<.05) and remained reduced with AT1 Ang II receptor blockade and rapid pacing (28.0+/ 0.5 microm/s, P<.05). Myocyte shortening velocity increased with ACEI or combination treatment compared with rapid pacing only (36.9+/-0.7 and 42.3+/-0.8 microm/s, respectively, P<.05). Myocyte beta-adrenergic response was reduced by >50% in both the rapid pacing group and the AT1 Ang II blockade group and improved by 25% with ACEI and increased by 54% with combined treatment. Both L type Ca2+ channel density and the relative abundance of sarcoplasmic reticulum Ca2+ ATPase density were reduced with rapid pacing and returned to control levels in the combined ACEI and AT1 Ang II blockade group. CONCLUSIONS: The unique findings of this study were twofold. First, basic defects in specific components of the myocyte excitation-contraction coupling process that occur with CHF are reversible. Second, combined ACEI and AT1 Ang II blockade may provide unique benefits on myocyte contractile processes in the setting of CHF. PMID- 9337217 TI - Endothelial dysfunction and cardiorenal injury in experimental salt-sensitive hypertension: effects of antihypertensive therapy. AB - BACKGROUND: Pharmacological control of hypertension has contributed to a significant decrease in cardiovascular morbidity and mortality, although the beneficial effect on cardiac and renal diseases has been far more modest than the reduction in stroke. The endothelium plays a crucial homeostatic role in the regulation of vascular tone thrombogenesis and vascular remodeling. We studied the relationship between endothelial dysfunction and cardiorenal injury in hypertensive rats and evaluated the effects of two classes of antihypertensive agents commonly used in the clinical setting, a diuretic (DIU) and an ACE inhibitor (CEI). METHODS AND RESULTS: Dahl salt-sensitive rats (DS) given high dietary salt (4% NaCl) developed hypertension (systolic blood pressure [SBP], 218+/-9 versus 147+/-3 mm Hg in DS given 0.5% NaCl; P<.001), which was associated with impaired endothelium-dependent relaxations (EDRs) in aortic rings (ED50, 5.44+/-.18 versus 7.51+/-.10; P<.05) and mesenteric vessels (area under the curve, 299+/-11 versus 217+/-11 arbitrary units; P<.05). Hypertensive DS also demonstrated depressed nitric oxide synthase activity in the aorta (0.76+/-.15 versus 2.83+/-.17 nmol x min(-1) x g protein(-1); P<.05), left ventricular hypertrophy (0.43+/-.02 versus 0.29+/-.02 g ventricular weight/100 g body weight; P<.05), glomerular injury (histological injury score: 151+/-8 versus 11+/-2; P<.05), and increased urinary protein excretion (95+/-21 versus 25+/-5 mg/24 hours; P<.05). Treatment of DS rats with the CEI perindopril (4.56 mg x kg(-1) x d(-1)) did not affect SBP (225+/-6 mm Hg) but modestly improved EDR (ED50: 6.07+/ .37; P<.05 versus hypertensive DS) as well as proteinuria and glomerular histology. Addition of the DIU indapamide (1.44 mg x kg(-1) x d(-1)) normalized SBP (151+/-2 mm Hg; P<.05), EDR (ED50, 7.33+/-.08; P<.05), left ventricular hypertrophy (0.27+/-.01 g/100 g body weight; P<.05), and proteinuria (31+/-4 mg/24 hours; P<.05) and prevented glomerular injury (injury score: 30+/-2; P<.05). Monotherapy with DIU reduced SBP (175+/-3 mm Hg; P<.05) and normalized EDR and left ventricular hypertrophy but did not provide effective renal protection. In hypertensive DS, impaired EDR and left ventricular hypertrophy were positively correlated with SBP. In addition, we found a significant correlation between cardiac hypertrophy and endothelial dysfunction. Indeed, a hierarchical regression analysis revealed that impaired aortic EDR, and therefore decreased aortic compliance, positively contributed to left ventricular hypertrophy in addition to but independently of SBP [F(2,37)=6.29; P=.004]. CONCLUSIONS: These studies suggest a dissociation of the endothelial, cardiac, and renal effects of antihypertensive therapy in hypertension and may explain the variable success of antihypertensive regimens in treating hypertension while preventing cardiac and renal damage. PMID- 9337218 TI - Right and left myocardial antioxidant responses during heart failure subsequent to myocardial infarction. AB - BACKGROUND: Heart failure subsequent to myocardial infarction (MI) is accompanied by depressed antioxidants and increased oxidative stress in the myocardium. Antioxidant enzyme activities and oxidative stress were examined in the viable left (LV) and right (RV) ventricles in relation to their hemodynamic function. METHODS AND RESULTS: The left coronary artery in rats was ligated. At 1 week after MI, LV systolic pressure (LVSP), LV end-diastolic pressure (LVEDP), and RV end-diastolic pressure (RVEDP) remained near control values, whereas RV systolic pressure (RVSP) was significantly elevated. In the 4, 8, and 16 week post-MI animals, LVSP was significantly reduced, with values of 112.0+/-1.57, 99.9+/ 0.52, and 89.2+/-1.4 mm Hg, whereas LVEDP was significantly elevated, with values of 8.2+/-0.52, 17.4+/-1.7, and 31.4+/-1.5 mm Hg, respectively. RVEDP was higher at 8 and 16 weeks, and RVSP was significantly reduced at 16 weeks. At 1 week after MI, myocardial catalase activity in the LV was maintained near control levels, whereas in the RV, it was 134% compared with its control value. At 4, 8, and 16 weeks, catalase activity in the LV was 71%, 48%, and 28% of respective controls. Catalase activity in the RV was significantly reduced only at 16 weeks. A similar trend was seen with respect to glutathione peroxidase activity. Reduced/oxidized glutathione ratio was significantly depressed in the LV at 1, 4, 8, and 16 weeks, whereas in the RV, this ratio was significantly reduced only at 8 and 16 weeks. Myocardial lipid peroxidation in the LV at 4, 8, and 16 weeks was elevated by approximately 40%, 51%, and 100%, respectively, whereas in the RV, an increase of approximately 50% was seen only at 16 weeks. CONCLUSIONS: These data show that heart failure subsequent to MI is associated with an antioxidant deficit as well as increased oxidative stress, first in the LV, followed by the RV. Furthermore, these changes correlated with the hemodynamic function in each of the ventricles, suggesting their role in the pathogenesis of ventricular dysfunction. PMID- 9337219 TI - Role of the Purkinje system in spontaneous ventricular tachycardia during acute ischemia in a canine model. AB - BACKGROUND: A role for the Purkinje system in the development of spontaneous ventricular tachycardia (VT) during acute ischemia has been suspected but not proved. We used a three-dimensional activation mapping system incorporating Purkinje signals to characterize the mechanism and site of origin of spontaneous VT occurring in the first 30 minutes after coronary artery occlusion in a dog model. METHODS AND RESULTS: The left anterior descending coronary artery was occluded in 48 dogs after instrumentation of the risk zone with 21 multipolar plunge needles, each recording 6 bipolar electrograms through the myocardial wall. VT of Purkinje origin was defined as a focal endocardial VT with a Purkinje potential identified before muscle potential on the electrode recording the earliest activity. Purkinje potentials were identified on an average of 10 of the 21 plunge needles. During atrial pacing at cycle lengths of 300 to 700 ms, a total of 25 VTs were observed from 18 of the 48 dogs (37.5%). Of the VTs, 15 (60.0%) were of focal Purkinje origin, 1 (4.0%) of focal endocardial origin, 2 (8.0%) of focal midmyocardial origin, and 2 (8.0%) of focal epicardial origin; 3 (12.0%) had a reentrant mechanism, whereas in 2 (8.0%), the mechanism could not be defined. The mean cycle length of all VTs was 265+/-17 ms (mean+/-SEM, n=25). Of the 25 VTs, 19 originated from an ischemic area as defined by significant decreases in voltages of muscle electrograms at the time of occurrence of the VT, 4 originated from an ischemic border zone, and the origin of 2 could not be determined. CONCLUSIONS: In this model, VT with a focal mechanism is commonly seen in the early ischemic period. Sixty percent of the VTs were of focal Purkinje origin as characterized by three-dimensional activation mapping. The results of this study indicate that Purkinje tissue may play an important role in the development of early ischemic VT. PMID- 9337220 TI - Transcutaneous multielectrode basket catheter for endocardial mapping and ablation of ventricular tachycardia in the pig. AB - BACKGROUND: Endocardial mapping using standard electrode catheters is often technically limited in ventricular tachycardia and constitutes a major obstacle to successful ablation. We wished to examine the utility of a basket-shaped multielectrode mapping catheter (MMC) in the mapping and ablation of ventricular tachycardia. METHODS AND RESULTS: This study of sustained monomorphic ventricular tachycardia (SMVT) was conducted in two phases in the postinfarction pig model. In the first phase, the utility of the MMC in providing adequate localization of potential ablation site(s) of SMVT by different techniques (presystolic potentials, pace mapping, and concealed entrainment) was assessed in 21 pigs. In the second phase, ablation of induced SMVT was attempted in 10 pigs. Mapping of SMVT was performed after percutaneous introduction of the MMC to the LV. Comprehensive mapping was performed in 90 episodes of SMVT and required 2.0 to 25 seconds. Diastolic potentials were recorded during 86 episodes; good or identical pace maps (> or = 9 of 12 paced surface ECG leads identical to ventricular tachycardia surface ECG leads) were obtained in 25 of 31 maps, and entrainment was achieved during 28 of 42 SMVTs. In 10 pigs, 10 SMVTs were recorded at least twice and were considered for radiofrequency ablation. An 8-mm tip ablation catheter was advanced to potential ablation sites with a specially designed "homing" device, requiring a median time of 120 seconds. In these 10 pigs, either identical pace map (> or = 11 of 12, 6 SMVTs) or concealed entrainment (4 SMVTs) guided the ablation procedure. After ablation, 8 of 10 SMVTs were rendered noninducible, while 2 pigs died during energy application of degeneration of SMVT to ventricular fibrillation. CONCLUSIONS: The MMC allows rapid, comprehensive, and reliable endocardial mapping during SMVTs, which facilitates successful ablation in the porcine post-myocardial infarction model. PMID- 9337221 TI - Percutaneous treatment of abdominal aortic aneurysm in a swine model: understanding the behavior of aortic aneurysm closure through a serial histopathological analysis. AB - BACKGROUND: Previous studies used covered stent grafts to treat abdominal aortic aneurysms; however, such devices block flow into aortic side branches. We used uncovered stents with and without additional embolization coils to treat abdominal aortic aneurysm in a swine model and examined serial histological changes in the aneurysms over a 6-month period. METHODS AND RESULTS: We examined aneurysms in 9 control and 9 treated pigs (5 received stents alone and 4 received stents and coils). Aneurysms were surgically created with abdominal fascia. Three days later, we percutaneously placed a self-expandable uncovered stent across the aneurysm. Coils were implanted through the stent into the aneurysm lumen. An aortogram immediately after stent placement showed no significant change in aneurysm lumen; however, in pigs that had aortograms between 6 weeks and 6 months after treatment, the diameter decreased (28% to 65%) in 4 of 5 pigs, and 1 had no discernible aneurysm. Three treated pigs died, but only 1 from rupture. In contrast, 7 untreated aneurysms ruptured (2 pigs died of other causes). Histological examination revealed that the aneurysm lumen was reduced after treatment by collagen production. This healing process was accelerated in aneurysms treated with both stents and coils. In contrast, only limited amounts of new collagen were found in untreated, ruptured aneurysms. Instead, the fascia was disrupted and there was evidence of collagen degradation. CONCLUSIONS: We found that uncovered stents reduce the likelihood of aneurysm rupture in a swine model without blocking arterial branches. The presence of coils enhanced filling of the lumen by collagen. PMID- 9337222 TI - Coronary risk estimation and treatment of hypercholesterolemia. AB - BACKGROUND: Evidence-based treatment of hypercholesterolemia currently recommended for rationalizing drug prescription requires justification of treatment by randomized trials, such as the West of Scotland Coronary Prevention Study (WOSCOPS) or the Scandinavian Simvastatin Survival Study (4S), and evaluation of its benefit from the estimation of the coronary risk of each patient. METHODS AND RESULTS: The latest European guidelines and Sheffield tables apply these principles and justify the decision to treat hypercholesterolemia if the Framingham coronary multivariate risk estimate is high enough, ie, >20% risk of coronary event at 10 years in the former and >1.5% risk of coronary death per year in the latter. Nevertheless, the practice of these two recent guidelines results in discrepancies in the decision to treat, because coronary morbidity was considered in one but mortality was considered in the other, and the risk required for treating may be extrapolated from different trials (4S or WOSCOPS). CONCLUSIONS: Although the principle of targeting lipid-lowering treatment to high risk subjects is unquestioned, further studies are needed to demonstrate that the Framingham risk profile is useful in selecting persons who are likely to benefit and to determine the place of newer risk factors and that of early noninvasive detection of atheroma in the risk estimation-based treatment. PMID- 9337223 TI - Pressor with promise: using vasopressin in cardiopulmonary arrest. PMID- 9337225 TI - Images in cardiovascular medicine. Radiation therapy-induced cardiac injury. PMID- 9337224 TI - Incidence of and risk factors for atrial fibrillation in older adults. AB - BACKGROUND: This study aimed to describe the incidence of atrial fibrillation (AF) among older adults during 3 years of follow-up. METHODS AND RESULTS: In this cohort study, 5201 adults > or = 65 years old were examined annually on four occasions between June 1989 and May 1993. At baseline, participants answered questionnaires and underwent a detailed examination that included carotid ultrasound, pulmonary function tests, ECG, and echocardiography. Subjects with a pacemaker or AF at baseline (n=357) were excluded. New cases of AF were identified from three sources: (1) annual self-reports, (2) annual ECGs, and (3) hospital discharge diagnoses. Cox proportional-hazards models were used to assess baseline risk factors as predictors of incident AF. Among 4844 participants, 304 developed a first episode of AF during an average follow-up of 3.28 years, for an incidence of 19.2 per 1000 person-years. The onset was strongly associated with age, male sex, and the presence of clinical cardiovascular disease. For men 65 to 74 and 75 to 84 years old, the incidences were 17.6 and 42.7, respectively, and for women, 10.1 and 21.6 events per 1000 person-years. In stepwise models, the use of diuretics, a history of valvular heart disease, coronary disease, advancing age, higher levels of systolic blood pressure, height, glucose, and left atrial size were all associated with an increased risk of AF. The use of beta-blockers and high levels of alcohol use, cholesterol, and forced expiratory volume in 1 second were associated with a reduced risk of AF. CONCLUSIONS: The incidence of AF in older adults may be higher than estimated by previous population studies. Left atrial size appears to be an important risk factor, and the control of blood pressure and glucose may be important in preventing the development of AF. PMID- 9337226 TI - Images in cardiovascular medicine. Demonstration of massive pulmonary embolism with spiral volumetric CT. PMID- 9337227 TI - Cardiovascular disease in women: a statement for healthcare professionals from the American Heart Association. Writing Group. PMID- 9337284 TI - Esophageal cancer: a case for aggressive staging and a tailored treatment plan. AB - A 57-year-old man with dysphagia was found to have a distal esophageal cancer. The tumor was staged radiographically (with endoesophageal ultrasonography) and operatively. The latter consisted of a thoracoscopic examination of the thoracic esophagus and surrounding lymph nodes, as well as a limited laparotomy to evaluate the stomach and the perigastric and celiac lymph nodes. It was determined that the patient had a high-risk lesion (locally advanced, T3-4 or N1). Induction chemotherapy with concurrent radiation therapy was administered. Following neoadjuvant treatment, the patient underwent an uneventful esophagectomy. No residual tumor was identified in the specimen. The patient was doing well 6 months following his surgery. This case demonstrates the use of staging to guide therapy in esophageal cancer. PMID- 9337285 TI - Endosonographic evaluation of the patient with esophageal cancer. AB - The prognosis for patients with carcinoma of the esophagus remains poor despite the recent use of aggressive combination therapies and radical surgical resection. Attempts to improve the survival of patients with esophageal carcinoma have been confounded by a lack of effective therapeutic options in the later stages of the disease and an inability to accurately identify the early disease stages. Endoscopic ultrasound is a novel technique that affords close-proximity imaging of the esophageal wall and its adjacent structures. Endosonography is superior to CT scan for assessing depth of tumor penetration (T stage) and lymph node status (N stage). Recent advances in endoscopic ultrasonography include the ability to perform ultrasound-guided fine-needle aspiration of mediastinal masses and lymph nodes. Therefore, endosonography is ideally suited for staging esophageal cancers. PMID- 9337286 TI - Minimally invasive staging for esophageal cancer. AB - Thoracoscopy is an excellent means for staging esophageal cancer. Staging of esophageal carcinoma facilitates prognostication and allocation of patients to appropriate treatment regimens. Thoracoscopy is also useful in biopsies of direct mediastinal invasion. Routine thoracoscopic and laparoscopic lymph node staging has been used in patients with esophageal carcinoma with excellent results. Thoracoscopy can allocate patients for neoadjuvant therapy and help avoid an unnecessary thoracotomy in patients found to have gross spread of locoregional disease. PMID- 9337287 TI - Multimodality therapy for esophageal cancer. AB - Over the past decade and a half, several strategies have been developed to improve the survival of patients with esophageal cancer. Two strategies employ either neoadjuvant chemotherapy or chemoradiotherapy followed by surgery to improve local-regional control and decrease the incidence of distant metastases. A third strategy uses nonsurgical therapy as definitive treatment for patients without metastatic disease. Single-institution pilot trials and randomized comparative trials have been conducted evaluating each approach. The rationale for these trials, results, and current recommendations are presented. PMID- 9337289 TI - Resection of non-small cell lung cancer: how much and by what route. AB - Surgical resection remains the preferred treatment, when possible, in patients with non-small cell lung cancer (NSCLC). A complete resection is required to potentially improve survival of these patients. Lobectomy is the minimum resection of choice. En bloc resections of involved adjacent organs and structures are performed routinely with acceptable morbidity and mortality. Mediastinal lymph node dissection allows accurate surgical and pathologic staging of lymph node disease but has yet to be proven efficacious as a curative procedure. The standard approach to the hemithorax is via posterolateral thoracotomy. Recent muscle-sparing incisions and video-assisted techniques have been employed safely to accomplish goals of surgery. This article evaluates past and current approaches to the resection of NSCLC, and looks at the impact of route and extent of resection on survival of NSCLC patients. PMID- 9337288 TI - The evolution of therapy for patients with stage IIIA (N2) lung cancer. AB - This is a demonstrable case report and discussion of recent trends in neoadjuvant therapy for patients with locally advanced stage IIIA (N2) non-small cell lung cancer. PMID- 9337290 TI - Preparing for pulmonary resection: preoperative evaluation of patients. AB - Preoperative evaluation of patients being considered for pulmonary resection is a common practice for both pulmonologists and internists. Traditionally, preoperative evaluation of this population has entailed identifying patients in whom pulmonary resection carries an unacceptably high risk of morbidity and mortality. However, recent advances in surgical technique and patient management have prompted a reconsideration of traditional preoperative approaches. This article reviews procedures currently used in the preoperative evaluation of patients considered for pulmonary resection, including the patient history, physical examination, and preoperative interventions, and addresses further evaluation of the high-risk patient. PMID- 9337291 TI - Current strategies for radiation therapy in non-small cell lung cancer. AB - Thoracic radiotherapy is widely used in patients with non-small cell lung cancer. Its role as adjuvant treatment before or after surgery has not been established clearly. In patients with locally advanced disease, the main cause of failure is the absence of local control. Recently, three treatment approaches have shown a beneficial effect on overall survival in randomized trials conducted in this group of patients: sequential combination of thoracic radiotherapy and cisplatin based chemotherapy, concomitant use of radiation and daily low-dose cisplatin therapy, and hyperfractionated accelerated radiotherapy. Another area that merits further investigation is the role of adjuvant surgery. PMID- 9337292 TI - Public health concerns about lung cancer: a case report. AB - An illustrative clinical case is presented and important features are highlighted regarding public health concerns about risk factors for lung cancer, lung cancer in women, and lung cancer screening. PMID- 9337293 TI - Measuring effectiveness of lung cancer screening: from consensus to controversy and back. AB - BACKGROUND: While intense controversy exists regarding screening for breast, colorectal, and prostate cancer, a consensus exists regarding lung cancer screening. All organizations recommend against any efforts to detect early lung cancer because each of four randomized controlled trials (RCTs) has failed to demonstrate a significant reduction in lung cancer mortality as a result of screening. SYNTHESIS: Disease-specific mortality is assumed to represent the best measure of screening effectiveness in RCTs, because it is not subject to confounding by lead time, length, or overdiagnosis biases. However, the effects of these biases are predictable, so accurate assessments of the degree of confounding by these biases can be made. Moreover, the ability of mortality to accurately reflect cancer death rates depends on the ability of randomization to create experimental and control populations that have an equal risk of dying of the disease under study, except insofar as early detection may reduce that risk. Because the majority of participants in screening trials never develop the disease under investigation, small absolute differences in disease risk between groups often persist despite randomization, and such differences translate into much larger proportional differences in the size of subgroups at risk for disease specific mortality. This effect confounds the ability of disease-specific mortality to accurately measure screening effectiveness. RESULTS: A total of 18 RCTs have been conducted to evaluate screening for breast, colorectal, and lung cancer. In the only two RCTs that reported a significant mortality reduction for screening mammography in breast cancer, and in the one RCT that reported a significant mortality reduction for fecal occult blood screening in colorectal cancer, population differences led mortality comparisons to overestimate the effectiveness of screening. In lung cancer, no significant mortality reductions have been reported (to my knowledge), but in the two RCTs most directly addressing the effectiveness of chest radiograph (CXR) screening, population differences led mortality comparisons to underestimate the ability of CXRs to reduce the risk of dying of lung cancer. Although mortality is believed to be the best measure of outcome, not a single example can be cited as definitive proof of efficacy for any screening strategy. Thus, screening cannot be recommended for any cancer on the basis of consistent reductions in mortality in RCTs. ANALYSIS: Current policy, which calls for no early detection efforts for lung cancer, implicitly accepts the validity of two contradictory assertions. Conventional wisdom maintains that lung cancer is a highly virulent disease and that metastases are present at inception; accordingly, early detection is ineffective. However, RCTs suggest that lung cancer is an indolent disease and that radiographically detected lesions are clinically unimportant; accordingly, early detection is unnecessary. Such contradictions mandate some rethinking of the fundamental assumptions underlying screening evaluation. CONCLUSIONS: Considerable evidence suggests that annual CXR screening could result in a dramatic reduction in lung cancer mortality in our society. However, proper interpretation of the data depends completely on how screening effectiveness is measured. Given the enormous public health importance of this issue, a consensus conference is recommended to determine whether lung cancer screening can save lives. PMID- 9337295 TI - Radiologic evaluation in lung cancer: diagnosis and staging. AB - Radiology and surgery are not competing but are complementary modalities in the care of patients with lung cancer. In certain areas, such as evaluation of the solitary pulmonary nodule, radiologic studies can have an important impact on patient care. Mediastinal staging with imaging studies is inexact, and CT may be most effective as a road map for more definitive surgical staging. MRI currently offers no advantages over CT in staging of the mediastinum but can be helpful in evaluation of parts of the chest not well demonstrated on axial images. A discussion of newer nuclear medicine imaging modalities is included. PMID- 9337294 TI - Women and lung cancer: waiting to exhale. AB - Lung cancer is now the leading cause of cancer deaths among women. In the United States, 64,300 women are expected to die of lung cancer in 1996. Smoking is responsible for about 80% of lung cancer cases. Unfortunately, the prevalence of smoking among women remains unacceptably high at about 22% and is expected to surpass the rate in men by the year 2000. Smoking rates are highest among young girls and the less educated. Whether lung cancer represents a different disease in women than in men is unclear. Data are conflicting regarding the magnitude of the relative risk of developing lung cancer due to smoking between the genders. There appears to be a difference in the relative distribution of lung cancer histologic features between men and women that is not explained entirely by differences in smoking patterns. Women who smoke appear to be at higher risk of developing small cell lung cancer than squamous cell lung cancer, whereas men who smoke have a similar risk for the two histologic conditions. Furthermore, women smokers are more likely to develop adenocarcinoma of the lung, and estrogens may play a causative role in this phenomenon. Data are unclear regarding whether the outcome of lung cancer treatment differs between genders. Solutions to the lung cancer epidemic among US women include (1) prevention of the disease by reducing smoking rates, (2) improving early detection methods, and (3) exploring new therapeutic strategies. PMID- 9337296 TI - Mediastinoscopy, thoracoscopy, and video-assisted thoracic surgery in the diagnosis and staging of lung cancer. AB - The intrathoracic staging of lung cancer involves assessment of the primary tumor and potential sites of metastases. Imaging studies of the chest are sensitive in detecting intrathoracic abnormalities, but specific staging information generally requires a tissue biopsy. The instruments used to obtain this information include the bronchoscope, mediastinoscope, and thoracoscope. The complementary application of these instruments can provide valuable staging information while limiting the morbidity of surgical staging. PMID- 9337297 TI - Implications of staging in lung cancer. AB - Lung cancer staging, based on anatomic extent of disease and described by the TNM staging system (T, primary tumor; N, regional lymph nodes; M, distant metastasis), is an important parameter for determining the clinical course of this disease. To evaluate the prognostic importance of TNM staging for lung cancer, we conducted a retrospective study analyzing survival rates according to TNM staging in 2,382 patients who had pulmonary resection for non-small cell lung cancer. Postoperatively, 3 patients were classified in stage 0, 796 in stage I, 304 in stage II, 719 in stage IIIA, 233 in stage IIIB, and 327 in stage IV. The 5 year survival rates for these patients were as follows: stage I, 68.5%; stage II, 46.9%; stage IIIA, 26.1%; stage IIIB, 9.0%; and stage IV, 11.2% (including ipsilateral, intrapulmonary metastases); 5-year survival rates for 140 patients with stage IV disease with intrapulmonary metastases in either the same lobe or another ipsilateral lobe were 17.8% and 8.3%, respectively. There was prognostic significance between stage I and stage II disease, stage II and stage IIIA disease, and stage IIIA and stage IIIB disease, but not between stage IIIB and stage IV disease. Only a few modifications will be required for the TNM staging system, which at present accurately reflects the prognosis of patients with lung cancer and is helpful in determining treatment. PMID- 9337298 TI - Limited-stage small cell lung cancer: a case report. AB - Because small cell lung cancer (SCLC) is very responsive to chemotherapy, an attempt at treatment is warranted even in poor-prognosis patients with limited stage disease. Concurrent thoracic radiotherapy and prophylactic cranial irradiation should be considered in such cases. A case report of an elderly, debilitated patient with limited-stage SCLC is presented, and his management is discussed. PMID- 9337299 TI - Small cell lung cancer: state-of-the-art therapy in 1996. AB - Small cell lung cancer (SCLC) occurs almost exclusively in smokers and represents 15 to 25% of all lung cancer histologic findings. It is distinguished from non small cell lung cancer by its rapid tumor doubling time, high growth fraction, and early development of widespread metastases. Since patients with SCLC usually present with disseminated disease, treatment strategies have focused on systemic therapy. Single-agent and combination chemotherapy, as well as combined-modality therapy, have produced high response rates (80 to 100% for limited disease; 60 to 80% for extensive disease), but these tend to be short-lived (median duration, 6 to 8 months). Survival beyond 5 years occurs in only 3 to 8% of all patients with SCLC. At least 15 to 20 different chemotherapeutic agents have shown major activity against SCLC in both the untreated and relapsed settings, including etoposide, teniposide, cisplatin, carboplatin, ifosfamide, cyclophosphamide, vincristine, and doxorubicin. This paper reviews state-of-the-art treatment strategies being employed in the treatment of SCLC, including those incorporating high-dose intensive therapy, salvage therapy, new agents, thoracic radiotherapy, prophylactic cranial radiotherapy, surgical resection, and biologic response modifiers. PMID- 9337300 TI - The role of thoracic radiotherapy in the management of limited-stage small cell lung cancer: past, present, and future. AB - The role of thoracic radiation therapy in the management of limited-stage small cell lung cancer (SCLC) is reviewed. Although chest irradiation has been used to treat SCLC for over four decades, its standard role in the management of limited stage disease was established only during the last decade. Multiple prospective randomized trials have shown that the addition of thoracic radiation therapy to chemotherapy usually halves local failure rates, from >60% with chemotherapy alone to about 30% with chemoradiation therapy. Additionally, survival at 3 years is also improved by 50%, from 10% with chemotherapy alone to about 15% with chemoradiation therapy. However, issues relating to the timing, volume (ie, prechemotherapy vs postchemotherapy), and the dose fractionation scheme of thoracic radiation therapy in the treatment of limited-stage SCLC still remain unresolved. Recent review of the literature indicates the most optimal timing of thoracic radiation therapy appears to be concurrent with chemotherapy vs either a sequential or an alternating approach. Studies are currently under way evaluating the optimal volume and dose fractionation scheme to use in the delivery of thoracic radiation therapy. In summary, thoracic radiation therapy significantly improves both local chest control and survival in the treatment of limited-stage SCLC. PMID- 9337301 TI - Non-small cell lung cancer: novel treatment strategies. AB - Prevention of cigarette smoking and early lung cancer detection remain important in our approach to the control of non-small cell lung cancer (NSCLC). In recent years, chemotherapy has emerged as a viable option in the treatment of NSCLC. The most impressive and widely confirmed evidence for this is the fact that chemotherapy can eradicate NSCLC micrometastases. Indeed, in some studies employing neoadjuvant chemotherapy followed by local surgery, pathology-confirmed complete remission rates as high as 20% have been reported. New agents showing preliminary activity in NSCLC include paclitaxel, vinorelbine, gemcitabine, and irinotecan (CPT-11). Certainly, however, there remains a need for novel, effective single-agent and combination chemotherapies. The seed/soil tumor concept, in which the seed consists of the tumor cells per se and the soil is the stroma containing the seeds, has proven helpful in devising new treatment strategies. Such strategies may include the use of antisoil agents, including antiangiogenesis, anti-invasion, and antimetastasis agents, both separately and particularly in conjunction with established antitumor agents. New therapeutic targets and methods of antitumor agent development based on modern molecular biology and pharmacology will provide a greater opportunity to improve the treatment of NSCLC. PMID- 9337302 TI - Innovative therapies for malignant pleural mesothelioma. AB - Therapy for malignant pleural mesothelioma is in a transitional stage. Recent trials of multimodality therapy for this disease suggest that selected patient subgroups may benefit from extensive treatment. This report discusses new approaches to the treatment of malignant pleural mesothelioma. Two case reports are presented. PMID- 9337303 TI - Multimodality therapy for malignant pleural mesothelioma. AB - Mesothelioma is a rare disease for which neither single modality nor bimodality therapy improves survival. For this reason, from 1980 to 1995, we used trimodality therapy in an attempt to improve survival in selected patients at Brigham and Women's Hospital and Dana-Farber Cancer Institute. One hundred twenty patients underwent trimodality treatment involving extrapleural pneumonectomy followed by combination chemoradiotherapy. Twenty-seven women and 93 men (mean age, 56 years) were evaluable for response and treatment-related morbidity. The operative mortality rate was 5%, and 22% of patients experienced major morbidity. Cell type and nodal status were significant prognostic variables. The respective 2- and 5-year survival rates were 45% and 22% overall, 70% and 37% for patients with epithelial cell type, 20% and 0% for patients with sarcomatous or mixed histologic-type tumors, and 74% and 39% for patients who were node-negative with epithelial histologic type. Positive resection margins impacted survival only in the case of full-thickness, transdiaphragmatic invasion. A revised staging system stratified survival with median intervals of 22, 17, and 11 months for stages I, II, and III disease, respectively (p=0.04). Thus, extrapleural pneumonectomy with adjuvant therapy is appropriate and effective treatment for patients with stage I disease according to the revised staging system. PMID- 9337304 TI - Brachytherapy for non-small cell lung cancer and selected neoplasms of the chest. AB - This article reviews the indications, techniques, and results of brachytherapy in the treatment of non-small cell lung cancer (NSCLC) and selected chest neoplasms. Various isotopes and techniques are used to place radioactive sources directly into a tumor, tumor bed, or the chest. Brachytherapy techniques can be tailored to the clinical situation and can be in the form of permanent interstitial volume or planar implants (radioactive sources permanently imbedded into the tumor or tumor bed) or in the form of temporary interstitial or endoluminal implants (where radioactive sources irradiate a tumor bed over a certain length of time and then are removed). These treatments can be delivered over a short interval (high-dose rate [HDR]) or over a more protracted time (low-dose rate). HDR treatments can be used intraoperatively to deliver a large dose of radiation to a determined target area with selective sparing of surrounding normal structures. Different methods of delivering HDR intraoperative radiation are under investigation. Most reports on brachytherapy for chest malignancies are retrospective and come from a few single institutions. Most of the published data relate to the treatment of NSCLC, but other intrathoracic malignancies, such as malignant pleural mesothelioma and malignant thymoma, have been treated with brachytherapy. To our knowledge, no major randomized trials accurately assess or confirm these retrospective studies yet, complicating the interpretation of these results. Nevertheless, brachytherapy is of value in selected situations and offers the clinician and the patient an innovative method of delivering conformal high-dose radiation to a defined target with preferential sparing of normal surrounding structures. With continued innovations in the development of radioactive isotopes, computerized treatment planning and targeting, and source delivery, brachytherapy should continue to offer an attractive alternative and complement to conventional treatment approaches, and may offer patients improved local control and survival. PMID- 9337305 TI - The surgical treatment of pulmonary metastases. AB - The surgical management of pulmonary metastases remains controversial, as no randomized trials have compared surgical excision with nonoperative treatment (to our knowledge). A Medline-generated review of the literature was undertaken to determine the factors influencing survival following metastasectomy in published trials. In the absence of randomized comparative trials, data must remain inferential and circumstantial. However, the literature does support the anecdotal observation that patients with metastatic disease can achieve long-term survival following surgical excision, irrespective of the source of the primary neoplasm, if there is no demonstrable extrathoracic disease and complete excision of the pulmonary disease is possible. Other factors noted as influencing survival appear to be anecdotal and variable from report to report. Pulmonary metastasectomy should be considered in patients with sufficient pulmonary reserve when the lung is the only site of metastatic disease and the lesions can be totally excised. An algorithm is proposed for a logical approach to the problem. PMID- 9337306 TI - Malignant effusive disease of the pleura and pericardium. AB - Malignant pleural and pericardial effusions are a common problem in the treatment of patients with lung cancer, breast cancer, or lymphoma and may occur with any malignancy. These effusions are frequently symptomatic and, in the case of the pleural space, may be the presenting sign of cancer. In other patients, they represent markers of recurrent, disseminated, or advanced disease. Given the poor prognosis of most patients presenting with these effusions, reducing symptoms and improving quality of life are the primary goals of treatment. Permanent drainage and/or obliteration of the pleural or pericardial space are crucial to the effective management of the effusion and will provide long-term palliation. Immediate relief can be accomplished via external drainage, but definitive therapy may often also require interventional radiology, cardiology, and thoracic surgery, as well as medical and radiation oncology. The pathophysiology, diagnosis, and treatment of malignant pleural and pericardial effusions are discussed in this article. PMID- 9337307 TI - Controversies in the management of malignant thymoma. AB - The management of most thymomas is relatively straightforward: surgical resection remains the primary mode of therapy. However, the literature contains many contradictory points of view regarding histology and pathology, staging and its usefulness, the need for adjuvant therapy, and recently, the place of video assisted surgery in the treatment of this tumor. This article is not a comprehensive guide to management but rather explores several of these controversial areas. Conclusions include the following: invasiveness remains the single most consistent factor in predicting outcome; surgery is the treatment of choice for thymoma whenever a complete resection can be accomplished; and incomplete resection may have some advantage over biopsy alone. The preponderance of evidence indicates that all thymomas except completely encapsulated stage I tumors should be treated with postoperative adjuvant radiation therapy in the hope of reducing the incidence of local relapse. Myasthenia can no longer be considered an adverse prognostic factor in thymoma; it may even confer a survival advantage, but this may be due to the preponderance of early-stage tumors discovered incidentally in myasthenic patients. Other associated autoimmune diseases confer a survival disadvantage. Demonstrating the equivalence of minimally invasive thoracoscopic approaches to standard thymectomy will take many years of investigation. Some promising reports on response to chemotherapy have led to the development of a phase II intergroup study to assess the value of chemotherapy in advanced thymoma. PMID- 9337337 TI - Micrometastasis of prostate cancer to lymph nodes: detection by means of reverse transcription-polymerase chain reaction. PMID- 9337338 TI - Anti-P-glycoprotein antibody C219 cross-reactivity with c-erbB2 protein: diagnostic and clinical implications. PMID- 9337339 TI - Can p53 status resolve paradoxes between human and non-human retinoblastoma models? PMID- 9337340 TI - Genes to order: regulators struggle against future abuses. PMID- 9337341 TI - Jekyll and Hyde: a new license for thalidomide? PMID- 9337342 TI - Vitamins during pregnancy linked to lower risk of childhood brain tumors. PMID- 9337343 TI - Solutions bring new problems: secondary cancers are a risk for some survivors. PMID- 9337344 TI - Texas center studies research alternative treatments. PMID- 9337345 TI - Aging and cancer: issues of basic and clinical science. AB - The majority of patients with cancer in the United States are more than 70 years old. Despite the increased understanding of the molecular bases for both oncogenesis and aging, the overlap of cancer and aging at that level remains a wide-open research domain. Similarly, at the clinical level, there is also an increased awareness of the need for more information about the influence of host age on the development of tumors, on the growth and spread of the disease, and on treatment expectations. In this review, we have attempted to frame questions regarding cancer and aging from the perspective of biogerontology and geriatric medicine. An increased effort to address the issues of aging is of paramount importance at all levels of cancer investigation. PMID- 9337346 TI - Prospective analysis of prostate-specific markers in pelvic lymph nodes of patients with high-risk prostate cancer. AB - BACKGROUND: Pathologic evidence of pelvic lymph node involvement is obtained in 12%-20% of patients with localized prostate cancer that exhibits high-risk features (defined on the basis of tumor size, serum prostate-specific antigen [PSA] level, or Gleason score). The rate of systemic failure (i.e., relapse) in patients with this type of prostate cancer and no pathologic evidence of regional lymph node involvement is 55%-92% within 5 years of definitive local therapy. Since reverse transcription-polymerase chain reaction (RT-PCR) methods are likely to be more sensitive than routine pathologic examination in detecting metastatic tumor cells, we compared the ability of the two approaches to detect prostate cells in the pelvic lymph nodes of patients with localized, high-risk disease. METHODS: Fifty-eight lymph node specimens isolated from 33 patients before definitive local therapy were examined. Expression of PSA and prostate-specific membrane antigen (PSM) messenger RNAs in the specimens was assessed by means of nested RT-PCR. RESULTS: Pathologic examination identified tumor cells in the lymph nodes of four (12%) of the 33 patients, and PSA and/ or PSM expression was positive in specimens from 27 (82%) of the patients (two-sided P<.0001). The four patients with positive pathologic findings also had positive RT-PCR results. Among the 29 patients with no pathologic evidence of lymph node involvement, 23 (79%) tested positive by means of RT-PCR. In these 23 patients, PSM expression was detected more frequently than PSA expression; however, in two patients, only PSA expression was detected. CONCLUSIONS: Expression of prostate-specific markers in the pelvic lymph nodes of patients with localized, high-risk prostate cancer may indicate the presence of metastatic tumor cells. Such cells may be responsible for the high rate of systemic failure seen in these patients. Additional studies are required to determine the prognostic relevance of our findings. PMID- 9337347 TI - Identification of epidermal growth factor receptor and c-erbB2 pathway inhibitors by correlation with gene expression patterns. AB - BACKGROUND: Growth factor receptor-signaling pathways are potentially important targets for anticancer therapy. The interaction of anticancer agents with specific molecular targets can be identified by correlating target expression patterns with cytotoxicity patterns. We sought to identify new agents that target and inhibit the activity of the epidermal growth factor (EGF) receptor and of c erbB2 (also called HER2 or neu), by correlating EGF receptor, transforming growth factor (TGF)-alpha (a ligand for EGF receptor), and c-erbB2 messenger RNA (mRNA) expression levels with the results of cytotoxicity assays of the 49000 compounds in the National Cancer Institute (NCI) drug screen database. METHODS: The levels of mRNAs were measured and used to generate a molecular target database for the 60 cell lines of the NCI anticancer drug screen. The computer analysis program, COMPARE, was used to search for cytotoxicity patterns in the NCI drug screen database that were highly correlated with EGF receptor, TGF-alpha, or c-erbB2 mRNA expression patterns. The putative EGF receptor-inhibiting compounds were tested for effects on basal tyrosine phosphorylation, in vitro EGF receptor tyrosine kinase activity, and EGF-dependent growth. Putative ErbB2-inhibiting compounds were tested for effects on antibody-induced ErbB2 tyrosine kinase activity. RESULTS: EGF receptor mRNA and TGF-alpha mRNA levels were highest in cell lines derived from renal cancers, and c-erbB2 mRNA levels were highest in cells derived from breast, ovarian, and colon cancers. Twenty-five compounds with high correlation coefficients (for cytotoxicity and levels of the measured mRNAs) were tested as inhibitors of the EGF receptor or c-erbB2 signaling pathways; 14 compounds were identified as inhibitors of these pathways. The most potent compound, B4, inhibited autophosphorylation (which occurs following activation) of ErbB2 by 50% in whole cells at 7.7 microM. CONCLUSIONS: Novel EGF receptor or c-erbB2 pathway inhibitors can be identified in the NCI drug screen by correlation of cytotoxicity patterns with EGF receptor or c-erbB2 mRNA expression levels. PMID- 9337348 TI - Cofactors with human papillomavirus in a population-based study of vulvar cancer. AB - BACKGROUND: Human papillomavirus (HPV) has been previously associated with vulvar cancer. In a population-based study, we examined whether exposure to HPV, cigarette smoking, or herpes simplex virus 2 (HSV2) increases the risk of this cancer. METHODS: Incident cases of in situ (n = 400) and invasive (n = 110) squamous cell vulvar cancer diagnosed among women living in the Seattle area from 1980 through 1994 were identified. Serum samples were analyzed for antibodies against specific HPV types and HSV2. HPV DNA in tumor tissue was detected by means of the polymerase chain reaction. In most analyses, case subjects were compared with population-based control subjects (n = 1403). Relative risks of developing vulvar cancer were estimated by use of adjusted odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Increased risks of in situ or invasive vulvar cancer were associated with HPV16 seropositivity (ORs = 3.6 [95% CI = 2.6 4.8] and 2.8 [95% CI = 1.7-4.7], respectively), current cigarette smoking (ORs = 6.4 [95% CI = 4.4-9.3] and 3.0 [95% CI = 1.7-5.3], respectively), and HSV2 seropositivity (ORs = 1.9 [95% CI = 1.4-2.6] and 1.5 [95% CI = 0.9-2.6], respectively). When the analysis was restricted to HPV16 DNA-positive tumors (in situ or invasive), the OR associated with HPV16 seropositivity was 4.5 (95% CI = 3.0-6.8). The OR for vulvar cancer was 18.8 (95% CI = 11.9-29.8) among current smokers who were HPV16 seropositive in comparison with never smokers who were HPV16 seronegative. CONCLUSIONS: Current smoking, infection with HPV16, and infection with HSV2 are risk factors for vulvar cancer. Risk appears particularly strong among women who are both current smokers and HPV16 seropositive. PMID- 9337349 TI - Cross-reactivity of C219 anti-p170(mdr-1) antibody with p185(c-erbB2) in breast cancer cells: cautions on evaluating p170(mdr-1) AB - BACKGROUND: Increased expression of the multidrug resistance gene (MDR-1)-encoded P-glycoprotein (p170[mdr-1]) is a major cause of tumor cell multidrug resistance. p170(mdr-1) functions as a drug-efflux pump to reduce the cellular accumulation of specific drugs. MDA-MB-435 human breast cancer cells that have been transfected with oncogene c-erbB2 complementary DNA (435.eb cells) express high levels of the transmembrane glycoprotein p185(c-erbB2) and exhibit increased resistance to the chemotherapeutic agent paclitaxel via p170(mdr-1)-independent mechanisms. We have recently discovered that the widely used monoclonal antibody C219, which is specific for p170(mdr-1), may cross-react with p185(c-erbB2) in 435.eb cells. In this study, we have investigated the nature of this cross reactivity. METHODS: Immunoprecipitation experiments involving the use of breast cancer cells that express different levels of p185(c-erbB2) were performed, and C219 was used for western blot analysis of immunoprecipitated proteins. Immunohistochemical analyses were performed on acetone-fixed slides of human breast cancer cells. Peptide sequence comparisons and enzyme-linked immunosorbent assays were performed to determine the molecular basis of C219 cross-reactivity with p185(c-erbB2). RESULTS: The cross-reactivity of C219 with p185(c-erbB2) was demonstrated by both western blot and immunohistochemical analyses. Peptide sequence comparisons revealed that C219 recognizes an epitope in p170(mdr-1) (C219 epitope) that shares sequence homology with p185(c-erbB2). Enzyme-linked immunosorbent assays demonstrated that C219 recognizes synthetic peptides derived from both the C219 epitope in p170(mdr-1) and the C219 epitope-homologous region in p185(c-erbB2). CONCLUSIONS: The anti-p170(mdr-1) monoclonal antibody C219 cross-reacts with p185(c-erbB2) through a peptide sequence in p185(c-erbB2) that is homologous to the C219 epitope in p170(mdr-1). PMID- 9337350 TI - Nuclear exclusion of wild-type p53 in immortalized human retinoblastoma cells. AB - BACKGROUND: Retinoblastoma is the most common childhood tumor of the eye, arising from cells that are defective in both copies of the retinoblastoma susceptibility gene (RB1). Most retinoblastoma tumor cells eventually undergo programmed cell death (i.e., apoptosis); however, some cells can acquire the ability to metastasize and become immortal. Transfection of immortal retinoblastoma cells with DNA sequences encoding wild-type p53 protein induces cell death, suggesting that the loss of both RB1 and p53 functions may be required for cell immortalization. We have examined this possibility by characterizing the p53 protein and messenger RNA in six independently isolated, immortalized retinoblastoma cell lines. METHODS: Western blotting methods were used to assess p53 protein level in each cell line, and Cleavase Fragment-Length Polymorphism analysis of complementary DNAs was used to screen for mutations in p53 messenger RNA. Localization of p53 protein in cells of the immortalized lines and in specimens of retinoblastoma tumors was achieved by means of indirect immunofluorescence and immunocytochemistry, respectively. RESULTS: All six immortalized cell lines expressed wild-type p53 messenger RNA and high levels of p53 protein. Although p53 is normally a nuclear protein, the p53 in four of the six cell lines was located predominately in the cytoplasm; in the remaining two cell lines, p53 was localized in both the nucleus and the cytoplasm. Cytoplasmic localization of p53 in retinoblastoma tumor specimens was rare and usually restricted to cells that had invaded adjacent ocular tissues, indicative of the early stages of metastasis. CONCLUSIONS: Some immortalized retinoblastoma cells may exhibit p53 dysfunction through nuclear exclusion of wild-type p53 protein. PMID- 9337351 TI - Loss of DNA mismatch repair: effects on the rate of mutation to drug resistance. AB - BACKGROUND: The loss of the ability of cells to repair mismatches in double stranded DNA is a common finding in human tumors. This defect results in genomic instability and in increased resistance to several of the drugs used in cancer chemotherapy. The human colon cancer cell line HCT116 is deficient in DNA mismatch repair (MMR) because of a genetic defect in the hMLH1 gene, which is located on chromosome 3. In this study, we investigated whether MMR-deficient HCT116+chr2 cells (i.e., HCT116 cells into which chromosome 2 has been transferred [as a control]) have a higher rate of mutation to resistance to commonly used chemotherapeutic agents (i.e., cisplatin, doxorubicin, paclitaxel [Taxol], and etoposide) than MMR-proficient HCT116+chr3 cells (i.e., HCT116 cells into which chromosome 3 has been transferred to provide a wild-type copy of the hMLH1 gene). METHODS: Spontaneous mutation rates were calculated from measurements of the mutant fractions of cells before and after their expansion through a known number of generations (also known as the technique of maximum likelihood estimation). Aliquots of 500000 cells were expanded in culture over a period of 2 weeks, and the mutant fractions were determined both before and after expansion of secondary cultures (each also with an initial 500000 cells) in drug concentrations that produced survival fractions of 0.0002%. RESULTS: Mutation rates in MMR-proficient and MMR-deficient cells did not differ on exposure to cisplatin, doxorubicin, or paclitaxel; however, the relative mutation rate was 2.4-fold higher in MMR-deficient cells exposed to etoposide (P=.002). CONCLUSION: These results suggest that genes involved in the control of cellular sensitivity to etoposide are targets for mutation when the loss of MMR destabilizes the genome. Tumors containing large fractions of MMR-deficient cells may demonstrate more rapid emergence of clinical resistance to etoposide. PMID- 9337352 TI - Age-specific incidence of acute lymphoblastic leukemia in U.S. children: in utero initiation model. PMID- 9337353 TI - Nitrate plasma level as a marker of nitric oxide production after subcutaneous interleukin 2 immunotherapy. PMID- 9337354 TI - Proposed modification of the "real Working" classification of lymphoid neoplasms. PMID- 9337355 TI - Re: prediction of carboplatin clearance from standard morphological and biological patient characteristics. PMID- 9337357 TI - Re: Oncologists judge themselves the best judges of cancer treatments. PMID- 9337356 TI - Identification of BRCA1 germline mutation, 797delAA, in a Japanese breast-ovarian cancer patient. PMID- 9337358 TI - Societal savings by "fast tracking" lower acuity patients in an urban pediatric emergency department. AB - To evaluate the cost-effectiveness of a "fast track" system for diverting lower acuity patients away from the pediatric emergency department (ED), 4,060 patients triaged to the fast track area of an urban pediatric ED with the 10 most common discharge diagnoses from 1/1/94 through 12/31/94 were retrospectively evaluated. Patients triaged as having nonurgent concerns qualified for treatment in a separate fast track area for 8 hours per day (fast track patients). These patients were compared with 5,199 seen in the main pediatric ED for the same concerns during the remaining hours when the fast track was not in operation (ED patients). Computer records were reviewed for demographics, acuity levels, diagnosis, and collection ratios (revenues/charges). The societal savings was calculated as sigma $ [(delta mean revenue of diagnosis1-10 in the main ED - mean revenue of diagnosis1-10 in the fast track) x the number of patients seen in fast track for diagnosis1-10] stratified by acuity. Collection ratios were comparable between groups (57% v 62%), but the average charges (physician and facility) were significantly less for patients seen in the fast track by a ratio of 1:2.4 (P < .0001). The average net revenue was also significantly less for all patients seen in the fast track by a ratio of 1:2.6 (P < .0001). When stratified by diagnosis and acuity, the savings to society was $101,313, or an average of $25/patient seen in the fast track ($101,313 per 4,060). A fast track is an effective system for maintaining patient flow at a cost savings to society. It can help the hospital in its negotiations with payors because it curtails charges. It is also a potential means for maintaining overall departmental revenues as payors increasingly deny traditional pediatric ED visits for patients with lower acuity concerns. PMID- 9337359 TI - Optimization of glottic exposure during intubation of a patient lying supine on the ground. AB - Two methods of endotracheal intubation of patients lying on the ground were compared for ease and speed of intubation and minimization of complications in a crossover study of prehospital-oriented emergency physicians. Intubation of a mannequin was attempted by the physicians in either a left lateral decubitus (LLD) position or a kneeling (K) position, followed by the alternate position. The LLD position afforded more rapid intubation, better glottic visualization, and less dental trauma. Eighty-seven percent of physicians completely visualized the glottis in the LLD position, versus 33% of the K position group. Intubation times were 10.5 versus 14.6 seconds in the LLD and K positions, respectively (P < .001). The LLD position is a more effective position (in a mannequin model) than the K position for intubation of patients found lying on the ground, a frequent situation in prehospital care. PMID- 9337360 TI - In-flight oral endotracheal intubation. AB - This study's goal was to analyze aeromedical emergency medical services (EMS) endotracheal intubation (ETI) success rates for in-flight intubations, and to retrospectively compare in-flight ETI success rates with those achieved in hospital and trauma scene settings. Patients undergoing flight crew ETI during a 3-year study period were reviewed, and flight team-performed intubations were classified as in-flight, hospital (at referring hospital), or ground (at trauma scene). Flight crews attempted ETI in 302 patients, with success in 291 patients (96.4%). ETI success rates for in-flight, hospital, and ground groups were 94.2%, 96.8%, and 98.3%, respectively (P = .22). There were no differences among the groups in proportions of pediatric patients (P = .55) or multiple intubation attempts (P = .83). Use of paralytic agents was more frequent in ground and in flight groups as compared with hospital group patients (P = .03). We conclude that with the aircraft and aeromedical crew studied, ETI was as likely to be successful in-flight as in other settings. PMID- 9337361 TI - Incidence of aspiration after urgent intubation. AB - This study sought to determine the incidence of aspiration after urgent endotracheal intubation (ET) performed in the emergency department (ED), and to offer a descriptive evaluation of these intubations. In a retrospective review of 133 charts, 87 patients met inclusion criteria. Aspiration occurred in 3 (3.5%) patients (95% confidence interval, 0%, 7.4%). One had witnessed aspiration, and 2 had positive sputum cultures. None of the 87 patients had a positive chest radiograph or unexplained hypoxemia up to 48 hours after ET. Rapid-sequence induction and oral ET was performed in 79 (91%) patients, whereas 4 spontaneously breathing patients were nasally intubated. Seventy percent of patients underwent ET by PGY I or II residents, 29% by PGY III or IV residents, and 1% by ED attending physicians. Seventy-seven patients were intubated on the first attempt, and airway blood or vomitus during ET was noted in 11 patients. This study offers significant descriptive information regarding urgent ET performed in the ED, and shows that aspiration after urgent ET occurs infrequently in ED patients. PMID- 9337362 TI - Emergency extracorporeal life support for patients with near-fatal status asthmaticus. AB - Extracorporeal life support (ECLS) was used to treat three patients with near fatal status asthmaticus who did not respond to aggressive medical therapies and mechanical ventilation under controlled permissive hypercapnia. ECLS was instituted in patient 1 because PaCO2 was excessively high and pH was excessively low, in patient 2 because hypoxemia and shock were not responsive to treatment, and in patient 3 because of sustained severe hypotension. ECLS supported adequate gas exchange until pulmonary function improved, diminishing the need for mechanical ventilation and preventing pulmonary complications. Pulmonary dysfunction improved markedly after only 21 to 86 hours of ECLS. Aggressive medical treatments were continued during ECLS. Our findings indicate that ECLS is a useful method for preventing death in patients with near-fatal status asthmaticus. PMID- 9337363 TI - Women's perception of pain and distress during intravenous catheterization and urethral mini-catheterization. AB - A study was done to prospectively compare the pain and distress of urethral mini catheterization (MC) with the pain and distress of intravenous (IV) catheterization in women. Ten-centimeter visual analog scales were used on which the left end represented "no pain" or "no distress" and the right "the worst pain imaginable" or "extreme distress." Distress was defined to include embarrassment, anxiety, or fear. A convenience sample of 40 adult women who presented to a university emergency department and were undergoing both procedures as part of their management appraised pain and distress after IV placement and urethral MC. The mean IV pain score was 4.2 cm, whereas the mean MC score was only 2.6 cm (P = .02). The IV and MC distress score means were similar (2.3 v 2.6 cm, P = .55). These results suggest that patients perceive urethral MC as less painful than IV placement. The distress scores of these two procedures did not differ and were low in both instances. PMID- 9337364 TI - Subcutaneous emphysema as an uncommon presentation of child abuse. AB - A case of subcutaneous emphysema and pneumomediastinum as a result of child abuse is presented to add to the spectrum of findings associated with child maltreatment. This case is a reminder that although most cases of subcutaneous emphysema resolve uneventfully, there still needs to be an aggressive search for a cause. In addition, in the pediatric age group, the history given should be carefully verified as being plausible because of the possibility that child abuse may be the true etiology. PMID- 9337365 TI - Profile of geriatric pelvic fractures presenting to the emergency department. AB - Few studies have examined differences in mechanism, presentation, and outcome of trauma in geriatric patients. This study compared pelvic fractures and associated injuries in geriatric and nongeriatric patients. The medical charts of all patients presenting to a large urban emergency medicine teaching program with a pelvic fracture between January 1, 1987 and December 31, 1993 were retrospectively reviewed by study-blinded physicians. Data collected included mechanism and site of injury, associated injuries, disposition (admission or discharge), need for operative repair, length of hospital stay, as well as subsequent deaths and causes. The data were stratified into patients less than 65 years of age (group A) and 65 years or older (group B). Two-hundred five pelvic fractures were reviewed with 85 (41%) in group B. A significantly greater number of pelvic fractures in group B occurred by fall (86% v 25%, P < .05) and significantly less by motor vehicle accident (14% v 75%, P < .05). Site of pelvic fracture differed significantly only in the decreased number of geriatric iliac fractures (6% v 16%, P < .05). The sites of pelvic fractures for geriatric patients in descending order were multiple sites (58%), pubic rami (56%), acetabulum (19%), ischium (11%), iliac (6%), and sacroiliac (2%), and did not differ from nongeriatric patients. Geriatric patients had significantly fewer total associated injuries (40% v 61%, P < .05) although associated chest injuries were significantly more common (21% v 8%, P < .05). Death occurred in three (3%) nongeriatric and nine (11%) geriatric patients. Six geriatric deaths were caused by exacerbation of underlying cardiovascular disease. Geriatric patients underwent significantly fewer operative procedures (6% v 43%, P < .05) but there were no significant differences in the percent admitted (85%) or mean length of hospital stay (9.59 days). Despite the decreased severity of pelvic fractures, care must be taken to prevent morbidity caused by exacerbation of premorbid illnesses in geriatric patients with pelvic fractures. PMID- 9337366 TI - Elder abuse and neglect: understanding the causes and potential risk factors. PMID- 9337367 TI - An unusual transcapitate fracture of the wrist. AB - A patient sustained a crush-type hyperextension injury to his wrist, and presented to the emergency department (ED) with wrist pain, swelling, and paresthesias along the median and superficial radial nerve distributions. His initial radiographic study was interpreted as showing a radial styloid fracture. The next day, he returned for a scheduled revisit with continued wrist pain and swelling. He also had loss of two-point discrimination in the median nerve distribution and loss of thumb opposition. Repeat radiographs were interpreted as showing not only a radial styloid fracture, but also a fracture of the capitate with the proximal fragment rotated 180 degrees in the sagittal plane. The patient was admitted for surgery, and did well, with good return of function. The unusual position of the capitate fracture obscured the common signs of fracture recognition and thus went unnoticed on the patient's initial ED visit. However, in light of the patient's disproportionate symptoms with seemingly negative diagnostic study results, appropriate follow-up care was given, and definitive treatment was appropriately rendered. PMID- 9337368 TI - Evaluation of cerebral hemodynamics in a head-injured patient with hypovolemia using transcranial Doppler sonography. AB - A 20-year-old man presented with hypovolemic shock caused by abdominal injury. Cerebral hemodynamics were evaluated by transcranial Doppler (TCD) sonography. Middle cerebral artery flow velocities decreased, and the pulsatility indices increased markedly. Particularly, the waveform of the left middle cerebral artery showed a systolic peak, suggesting an increased intracranial pressure. Actual intracranial pressure was 7 mm Hg, and the cerebral perfusion pressure (CPP) was 51 mm Hg. These abnormal Doppler signals seemed to be caused by a compromise in CPP and to be aggravated by hypovolemia. The patient was discharged with a residual mild memory disturbance. Hypovolemia aggravates a reduced cerebral blood flow caused by a compromised CPP, and the waveform of TCD in a case of hypovolemic shock should be differentiated from intracranial hypertension. PMID- 9337369 TI - Addressing the myths of cervical spine injury management. AB - Every year in the United States about 5,000 people sustain a cervical spinal cord injury. Vastly greater numbers present to hospitals after motor vehicle crashes and falls with potential cervical spine injuries (CSI) for evaluation. This group of patients requires very careful management while undergoing evaluation for potential CSI to minimize the potential for spinal cord injury. It is, therefore, incumbent on everyone caring for these patients to distinguish between fact and fiction in regard to CSI management. This article addresses the following areas of controversy: CSI is a rare injury; patients with cranial and facial injuries are at increased risk for CSI; everyone with a significant mechanism of injury needs radiological clearance of their cervical spine; a normal cross-table lateral view radiograph excludes significant CSI; oral intubation of patients with CSI is not safe; a semi-rigid collar prevents movement of the cervical spine; and the evaluation of the cervical spine needs to begin in the resuscitation room in every patient. PMID- 9337370 TI - The dynamics of patient visits to a public hospital ED: a statistical model. AB - Using a public hospital's computerized database, we formulated a statistical model to explain emergency department (ED) patient volume for better staffing and resource allocation. All patients visiting the ED over a 3-year period were included in this retrospective study. Each observation described the total daily number of referrals and was defined by the following variables: day of the week, month of the year, holiday/ weekday, relative order in a 3-year sequence, and number of visits to the ED on that day. The statistical method used to build the model was analysis of covariance. Periodicity in average number of daily visits existed for each of the seasonal factors that were examined, repeating every year during the study period. Based on a graphic analysis, the model was defined and explained 65% of the variance during the 3-year study, with a relatively low standard deviation of error. A statistically significant correlation existed between time-related factors and the number of visits to the ED. This statistical model may prove to be of value for planning emergency services, which operate under stressful, unpredictable situations. PMID- 9337372 TI - Trauma: an annotated bibliography of the recent literature. PMID- 9337371 TI - Designing a prehospital system for a developing country: estimated cost and benefits. AB - Many of the costs associated with prehospital care in developed countries are covered in budgets for fire suppression, police services, and the like. Determining these costs is therefore difficult. The costs and benefits of developing a prehospital care system for Kuala Lumpur, Malaysia, which now has essentially no emergency medical services (EMS) system, were estimated. Prehospital therapies that have been suggested to decrease mortality were identified. A minimal prehospital system was designed to deliver these treatments in Kuala Lumpur. The potential benefit of these therapies was calculated by using statistics from the United States corrected for demographic differences between the United States and Malaysia. Costs were extrapolated from the current operating budget of the Malaysian Red Crescent Society. Primary dysrhythmias are responsible for almost all potentially survivable cardiac arrests. A system designed to deliver a defibrillator to 85% of arrests within 6 minutes would require an estimated 48 ambulances. Kuala Lumpur has approximately 120 prehospital arrhythmic deaths per year. A 6% resuscitation rate was chosen for the denominator, resulting in seven survivors. Half of these would be expected to have significant neurological damage. Ambulances cost $53,000 (US dollars) to operate per year in Kuala Lumpur; 48 ambulances would cost a total of $2.5 million. Demographic factors and traffic problems would significantly increase the cost per patient. Other therapies, including medications, airway management, and trauma care, were discounted because both their additional cost and their benefit are small. Transport of patients (including trauma) is now performed by police or private vehicle and would probably take longer by ambulance. A prehospital system for Kuala Lumpur would cost approximately $2.5 million per year. It might save seven lives, three of which would be marred by significant neurological injury. Developing countries would do well to consider alternatives to a North American EMS model. PMID- 9337374 TI - Prehospital use of intravenous diltiazem (cardizem Lyo-Ject) in the treatment of rapid atrial fibrillation. PMID- 9337373 TI - Self-inflicted burn injury. PMID- 9337375 TI - Simultaneous bilateral femoral-neck fractures in an elderly woman. PMID- 9337376 TI - Efficacy of the rhesus rotavirus-based quadrivalent vaccine in infants and young children in Venezuela. AB - BACKGROUND: Rotaviruses are the principal known etiologic agents of severe diarrhea among infants and young children worldwide. Although a rhesus rotavirus based quadrivalent vaccine is highly effective in preventing severe diarrhea in developed countries, in developing countries its efficacy has been less impressive. We thus conducted a catchment study in Venezuela to assess the efficacy of the vaccine against dehydrating diarrhea. METHODS: In this randomized, double-blind, placebo-controlled trial, 2207 infants received three oral doses of the quadrivalent rotavirus vaccine (4x10(5) plaque-forming units per dose) or placebo at about two, three, and four months of age. During approximately 19 to 20 months of passive surveillance, episodes of gastroenteritis were evaluated at the hospital. RESULTS: The vaccine was safe, although 15 percent of the vaccinated infants had febrile episodes (rectal temperature, > or =38.1 degrees C) during the six days after the first dose, as compared with 7 percent of the controls (P<0.001). However, the vaccine gave 88 percent protection against severe diarrhea caused by rotavirus and 75 percent protection against dehydration, and produced a 70 percent reduction in hospital admissions. Overall, the efficacy of the vaccine against a first episode of rotavirus diarrhea was 48 percent. Horizontal transmission of vaccine virus was demonstrated in 15 percent of the vaccine recipients and 13 percent of the placebo recipients with rotavirus-positive diarrhea. CONCLUSIONS: In this study in a developing country, the quadrivalent rhesus rotavirus-based vaccine induced a high level of protection against severe diarrheal illness caused by rotavirus. PMID- 9337377 TI - Prognostic value of immunohistochemically identifiable tumor cells in lymph nodes of patients with completely resected esophageal cancer. AB - BACKGROUND: Current methods of disease staging often fail to detect small numbers of tumor cells in lymph nodes. Metastatic relapse may arise from these few cells. METHODS: We studied 1308 lymph nodes from 68 patients with esophageal cancer without overt metastases who had undergone radical en bloc esophagectomy. A total of 399 lymph nodes obtained from 68 patients were found to be free of tumor by routine histopathological analysis and were studied further for isolated tumor cells by immunohistochemical analysis with the monoclonal anti-epithelial-cell antibody Ber-EP4. This antibody did not stain lymph nodes from 24 control patients without carcinoma. RESULTS: Of the 399 "tumor free" lymph nodes, 67 (17 percent), obtained from 42 of the 68 patients, contained Ber-EP4-positive tumor cells. Fifteen of 30 patients who were considered free of lymph-node metastases by histopathological analysis had such cells in their lymph nodes, and 5 of the 15 had small primary tumors. Ber-EP4-positive cells found in "tumor free" nodes were independently predictive of significantly reduced relapse-free survival (P=0.008) and overall survival (P=0.03). They predicted relapse both in patients without nodal metastases (P=0.01) and in those with regional lymph-node involvement (P=0.007). All 12 patients whose lymph nodes were negative on both histopathological and immunohistochemical analysis and who were available for follow-up survived without recurrence. The presence of micrometastatic tumor cells in bone marrow had no additional prognostic value. CONCLUSIONS: Immunohistochemical examination of lymph nodes may improve the pathological staging of esophageal cancer. PMID- 9337378 TI - A comparison of sustained-release bupropion and placebo for smoking cessation. AB - BACKGROUND AND METHODS: Trials of antidepressant medications for smoking cessation have had mixed results. We conducted a double-blind, placebo-controlled trial of a sustained-release form of bupropion for smoking cessation. We excluded smokers with current depression, but not those with a history of major depression. The 615 subjects were randomly assigned to receive placebo or bupropion at a dose of 100, 150, or 300 mg per day for seven weeks. The target quitting date (or "target quit date") was one week after the beginning of treatment. Brief counseling was provided at base line, weekly during treatment, and at 8, 12, 26, and 52 weeks. Self-reported abstinence was confirmed by a carbon monoxide concentration in expired air of 10 ppm or less. RESULTS: At the end of seven weeks of treatment, the rates of smoking cessation as confirmed by carbon monoxide measurements were 19.0 percent in the placebo group, 28.8 percent in the 100-mg group, 38.6 percent in the 150-mg group, and 44.2 percent in the 300-mg group (P<0.001). At one year the respective rates were 12.4 percent, 19.6 percent, 22.9 percent, and 23.1 percent. The rates for the 150-mg group (P=0.02) and the 300-mg group (P=0.01) -- but not the 100-mg group (P=0.09) -- were significantly better than those for the placebo group. Among the subjects who were continuously abstinent through the end of treatment, the mean absolute weight gain was inversely associated with the dose (a gain of 2.9 kg in the placebo group, 2.3 kg in 100-mg and 150-mg groups, and 1.5 kg in the 300-mg group; P= 0.02). No effects of treatment were observed on depression scores as measured serially by the Beck Depression Inventory. Thirty-seven subjects stopped treatment prematurely because of adverse events; the frequency was similar among all groups. CONCLUSIONS: A sustained-release form of bupropion was effective for smoking cessation and was accompanied by reduced weight gain and minimal side effects. Many participants in all groups were smoking at one year. PMID- 9337379 TI - Fasting hypoketotic coma in a child with deficiency of mitochondrial 3-hydroxy-3 methylglutaryl-CoA synthase. PMID- 9337380 TI - Images in clinical medicine. A little survivor. PMID- 9337381 TI - Differing birth weight among infants of U.S.-born blacks, African-born blacks, and U.S.-born whites. AB - BACKGROUND: In the United States, the birth weights of infants of black women are lower than those of infants of white women. The extent to which the lower birth weights among blacks are related to social or genetic factors is unclear. METHODS: We used vital records for 1980 through 1995 from Illinois to determine the distribution of birth weights among infants born to three groups of women -- U.S.-born blacks, African-born blacks, and U.S.-born whites. RESULTS: The mean birth weight of 44,046 infants of U.S.-born white women was 3446 g, that of 3135 infants of African-born black women was 3333 g, and that of 43,322 infants of U.S.-born black women was 3089 g. The incidence of low birth weight (weight less than 2500 g) was 13.2 percent among infants of U.S.-born black women and 7.1 percent among infants of African-born black women, as compared with 4.3 percent among infants of U.S.-born white women (relative risks, 3.1 and 1.6, respectively). Among the women at lowest risk (those 20 to 39 years old, with 12 years of education for themselves and their spouses, early prenatal care, gravida 2 or 3, and no previous fetal loss), the rate of low birth weight in infants of African-born black women (3.6 percent) was closer to the rate in infants of U.S. born white women (2.4 percent), and the rate in infants of U.S.-born black women remained high (7.5 percent). CONCLUSIONS: The birth-weight patterns of infants of African-born black women and U.S.-born white women are more closely related to one another than to the birth weights of infants of U.S.-born black women. PMID- 9337382 TI - Anesthesiology. Second of two parts. PMID- 9337383 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 33-1997. A 75-year-old man with chest pain, hemoptysis, and a pulmonary lesion. PMID- 9337384 TI - A vaccine against rotavirus--when is too much too much? PMID- 9337385 TI - Treating tobacco addiction--nicotine or no nicotine? PMID- 9337386 TI - The enigma of low birth weight and race. PMID- 9337388 TI - The Supreme Court speaks--not assisted suicide but a constitutional right to palliative care. PMID- 9337389 TI - Mutation watch: mouse brachyury (T), the T-box gene family, and human disease. PMID- 9337390 TI - Pas1 is a common lung cancer susceptibility locus in three mouse strains. AB - Inherited predisposition to lung cancer is a phenotypic trait shared by different mouse inbred strains that show either a high or an intermediate predisposition. Other strains are instead genetically resistant. The Pas1 locus is the major determinant of lung cancer predisposition in the A/J strain (Gariboldi et al. 1993). To define the determinants of susceptibility to lung tumorigenesis in the highly susceptible SWR/J and in the intermediately susceptible BALB/c mice, we analyzed (BALB/c x SWR/J)F2 and (BALB/c x C3H/He)F2 crosses by genetic linkage experiments. The present results provide unequivocal evidence that the same Pas1/+ allele that leads to lung cancer predisposition is shared by A/J, SWR/J, and BALB/c strains. The intermediate susceptibility of the BALB/c strain would result by interaction of Pas1 locus with lung cancer resistance loci. PMID- 9337392 TI - Steroid hormone responsiveness of a family of closely related mouse proviral elements. AB - Regulation of the mouse sex-limited protein (Slp) gene in unusual in that hormone response is conferred by the 5' LTR of an upstream inserted provirus, dubbed the imposon (imp1). In a search for additional genes whose regulation has been affected by retrotransposition events, we isolated two partial proviral elements by stringent screening of a mouse genomic library. One clone (imp2) contained a portion of the envelope gene and a 3' LTR that was nearly identical to the 3' LTR of imp1; this similarity extended to insertion into a B1 repetitive element. The second proviral clone (imp3) contained a 5' LTR and associated coding sequences, but lacked its 3' LTR; the LTR of imp3 differed by 12% from the imp1 sequence. To assess potential hormone response, proviral enhancer regions cloned into reporter vectors were tested in transfection. The imp2 enhancer was similar in behavior to imp1, conferring both androgen and glucocorticoid induction in one fragment context and an androgen-specific response in another. In contrast, the imp3 enhancer allowed high expression in the absence of hormone and was less responsive to steroids in general and androgen in particular. These three proviral elements define a small family of steroid responsive proviruses in the mouse genome, and at least one member has had a lasting impact on an endogenous gene's regulation. PMID- 9337391 TI - Matrix-attachment regions in the mouse chromosome 7F imprinted domain. AB - We have mapped the matrix-attachment regions (MARs) in 200 kilobases of the mouse Chromosome (Chr) 7F imprinted domain. MARs are genetic elements known to have effects in cis on methylation at nonimprinted loci. The imprinting of the Igf2 and Ins2 genes is dependent on the transcription of the downstream H19 gene. The transcription of H19 is dependent in turn on its methylation status. The cis acting regulators of methylation at this site are not known. As MARs are potential regulators not only of methylation but also other elements of genomic imprinting, we mapped the MARs within the 200 kilobases around H19. This report describes the mapping of four MARs from this region. PMID- 9337393 TI - Coordinate control and variation in X-linked gene expression among female mice. AB - In normal female mammals, one of the two X Chromosome (Chr) homologs per cell is silenced coordinately during early embryogenesis. The genes located on the inactivated X homolog are predicted to be influenced by the same underlying repression mechanism. To test the uniformity of cis-acting gene repression, 32 genetically identical F1 female mice were analyzed for differential expression of homologous alleles at three X-linked genes-Otc, Atp7a (= Mottled), and Hprt. Gene expression was assayed by the single-nucleotide primer extension (SNuPE) method, thereby allowing the three genes to be quantitated from the same RNA sample. Although variable between individual animals, the relative expression of the two alleles (allelic expression ratio) of the genes is significantly correlated within each steady-state RNA pool. When examined by animal age (3 months to 12 months), no statistically significant differences were observed in the mean or variance of allelic expression ratio. Together, the results confirm that X inactivation is coordinately controlled and is stable across the early- to mid adult life span. PMID- 9337394 TI - Quantitative trait loci for estrogen-dependent pituitary tumor growth in the rat. AB - Growth control is of fundamental importance to biology in general and of critical importance to cancer research in particular. Tumors develop when control of the normal growth process is lost. The rat pituitary is a model system for control of estrogen-dependent growth. Chronic estrogen treatment induces uncontrolled growth in the pituitaries of Fischer 344 (F344) rats, but not of Brown Norway (BN) rats. We have identified five quantitative trait loci (QTL) for estrogen-dependent pituitary mass (Edpm) in an F2 intercross of F344 and BN. These QTL reside on rat Chromosomes (Chrs) 2, 3, 5, and 9 and explain a total of 55% of the genetic variance in the F2. We have also detected suggestive evidence for a QTL on rat Chr 14. For Edpm2-1, Edpm2-2, Edpm3, and Edpm5, the F344 allele corresponds with increased pituitary mass, as expected. Surprisingly, for Edpm9 and the suggested QTL on Chr 14, the BN allele corresponds with increased pituitary mass. We also find evidence for interaction (epistasis) between Edpm3 and Edpm9 and between Edpm5 and the suggested QTL on Chr 14. PMID- 9337396 TI - Targeted development of microsatellite markers from the defined region of bovine chromosome 6q21-31. AB - A methodical strategy for the isolation of microsatellite markers specific for targeted regions of bovine chromosomes is presented. The procedure involves directed microdissection of one defined subchromosomal area, its DOP-PCR amplification and cloning. With this approach, a library specific to the BTA 6q21 31 chromosomal region was constructed. Eleven unique microsatellite-containing sequences were isolated, converted into sequence-tagged microsatellite sites, and characterized concerning their species-specific origin. Seven primer pairs generated bovine-specific PCR products and provided a set of microsatellite markers that generally revealed high informativity in the HF breed. Linkage analysis assigned six of them to their predefined subchromosomal origin on BTA 6 corresponding to the specific rehybridization signal of the DOP-PCR product generated from the microdissected chromosome area 6q21-31. The results underline the usefulness of the BTA 6q21-31 library for targeted isolation of unique sequences that are specific for the dissected chromosomal region as demonstrated here by the isolation of microsatellite markers. PMID- 9337395 TI - Evidence for individual and between-family variability of the recombination rate in cattle. AB - We have conducted a study based on single sperm typing in a family design to assess patterns of variability of the male recombination rate in cattle. 2214 sperm of 37 bulls were typed for 11 loci on bovine Chromosomes (Chrs) 6, 23, and the sex chromosomes. Statistically significant individual variability of the recombination rate was observed for one interval in the pseudoautosomal region (PAR) of the bovine sex chromosomes; one marker interval on bovine Chr 23 exhibited individual variability that was close to significance. Thirty-five of the bulls were members of six paternal halfsib groups, and highly significant variability between families was found for one interval in the PAR. This variability may be due to DNA sequence differences in the PAR or to a genetic control of the recombination activity in this region. It is demonstrated that differences in the recombination rate of the magnitude observed in the present study may have a considerable impact on the power of QTL mapping experiments as well as on the sustainability of marker-assisted selection strategies. PMID- 9337397 TI - Human and murine PTX1/Ptx1 gene maps to the region for Treacher Collins syndrome. AB - Ptx1 belongs to an expanding family of bicoid-related vertebrate homeobox genes. These genes, like their Drosophila homolog, seem to play a role in the development of anterior structures and, in particular, the brain and facies. We report the chromosomal localization of mouse Ptx1, and the cloning, sequencing, and chromosomal localization of the human homolog PTX1. The putative encoded proteins share 100% homology in the homeodomain and are 88% and 97% conserved in the N- and C-termini respectively. Intron/exon boundaries are also conserved. Murine Ptx1 was localized, by interspecific backcrossing, to Chr 13 within 2.6 cM of Caml. The gene resides centrally on Chromosome (Chr) 13 in a region syntenic with human Chr 5q. Subsequent analysis by fluorescent in situ hybridization places the human gene, PTX1, on 5q31, a region associated with Treacher Collins Franceschetti Syndrome. Taken together with the craniofacial expression pattern of Ptx1 during early development, the localization of the gene in this chromosomal area is consistent with an involvement in Treacher Collins Franceschetti Syndrome. PMID- 9337398 TI - An additional 150 SSLP markers typed for the AXB and BXA recombinant inbred mouse strains. PMID- 9337399 TI - Cloning of rat Brca2 and linkage mapping to chromosome 12. PMID- 9337400 TI - Mapping of the ATP2B2 and PCCB genes on porcine chromosome 13. PMID- 9337402 TI - Allele-specific PCR assays for the tub and cpefat mutations. PMID- 9337403 TI - Expressed sequences within pericentromeric heterochromatin of human chromosome 22. PMID- 9337401 TI - A whole-genome radiation hybrid panel for bovine gene mapping. PMID- 9337404 TI - Cloning, chromosomal mapping, and expression of the human eHAND gene. PMID- 9337405 TI - Cloning and mapping of the human SOX1: a highly conserved gene expressed in the developing brain. PMID- 9337406 TI - Assignment of the murine Hmx1 homeobox gene to the proximal region of mouse chromosome 5. PMID- 9337408 TI - Regional localization of the mouse argininosuccinate lyase gene to chromosome 5. PMID- 9337407 TI - Mapping of the gene for calreticulin (Calr) to mouse chromosome 8. PMID- 9337409 TI - Localization of a neural crest transcription factor, Slug, to mouse chromosome 16 and human chromosome 8. PMID- 9337410 TI - Mapping of six germ-cell-specific genes to mouse chromosomes. PMID- 9337412 TI - Survey of Culturable Airborne Bacteria at Four Diverse Locations in Oregon: Urban, Rural, Forest, and Coastal PMID- 9337411 TI - Galanin receptor 1 gene (Galnr1) is tightly linked to the myelin basic protein gene on chromosome 18 in mouse. PMID- 9337413 TI - Hybridization Analysis of Microbial DNA from Fuel Oil-Contaminated and Noncontaminated Soil PMID- 9337414 TI - Microbial Food Webs in Marine Sediments. I. Trophic Interactions and Grazing Rates in Two Tidal Flat Communities PMID- 9337415 TI - Microbial Food Webs in Marine Sediments. II. Seasonal Changes in Trophic Interactions in a Sandy Tidal Flat Community AB - >AbstractThe role of grazing by marine sediment flagellates, ciliates, and meiobenthic animals in controlling production of their bacterial and diatom prey was investigated. At six selected time points, over the year, bacterial production and diatom standing stock were compared to grazing pressure exercised by proto- and micrometazoan consumers. The intensity of prey-predator relations showed pronounced yearly dynamics in which two stages could be distinguished. During the first phase, from the end of winter to mid-summer, the consumption of diatoms gradually increased, with possible overgrazing at the end of the period. This was followed by a collapse of diatom abundance, to the winter level. During the first stage, no appreciable bacterial consumption was observed in spite of the high abundance and production of bacteria. The second stage started in mid summer and continued through the fall. During this period, the grazing on bacteria increased and reached the year's maximum. For at least a brief period (October), micrograzers removed the majority of bacterial production. In contrast, herbivory stayed at the year's lowest level, and diatoms appeared to be controlled by factors other than grazing. The observed ingestion rates seem to support the apparent energy requirements of flagellates and some ciliates (scuticociliates and hypotrichids). Other ciliates (pleurostomatids and karyorelictids) could not subsist on the observed diet and might have to complement it with other energy sources, possibly via dissolved organic matter absorption. PMID- 9337416 TI - Temporal Changes in the Bacterial Communities of Soil, Rhizosphere, and Endorhiza Associated with Field-Grown Cucumber (Cucumis sativus L.) PMID- 9337417 TI - The Response of Three Bacterial Populations to Pollution in a Stream PMID- 9337419 TI - The Relationship Between Electron Transport Activity as Measured by CTC Reduction and CO2 Production in Mixed Microbial Communities PMID- 9337418 TI - Saprospira grandis: A Flexibacterium That Can Catch Bacterial Prey by "Ixotrophy" PMID- 9337420 TI - Lack of Capsular Exopolymer Effects on the Biodegradation of Organic Compounds by Pseudomonas sp. Strains JS1 and JS150 PMID- 9337421 TI - The Fate of 15N-Nitrate in Healthy and Declining Phragmites australis Stands PMID- 9337422 TI - Update on mast cells and mast cell precursors and hypersensitivity responses. AB - Mast cells are important effector cells providing granule and membrane mediators as well as cytokines in allergic and inflammatory diseases. The study of surface molecules such as immunoglobulin receptors and adhesion molecules has greatly expanded the functional implications of mast cells. An active role for mast cells in antigen presentation to T cells has recently been shown, and direct interaction between mast cells and B cells providing signals for specific IgE production has been demonstrated. Functional receptors other than the high affinity IgE (Fc epsilon RI) have been implicated in the anaphylactic response of IgE-deficient mice, suggesting that IgG receptors present in mast cells may be involved in immediate hypersensitivity reactions. Although metachromatic mast cells are easily recognized in peripheral tissues, little is known about the phenotype of mast cell precursors, their fate from the bone marrow to the tissues, migration and homing processes, and factors and adhesion molecules that affect those processes. This review will describe the most recent studies in mouse and human mast cell biology and ontogeny. PMID- 9337423 TI - The pathobiology of eosinophilic inflammation. AB - Eosinophils are bone-marrow-derived granulocytes that are involved in both allergic and nonallergic inflammation. They possess a diverse repertoire of functional responses and effector capabilities, including the release of preformed cytotoxic granule proteins, superoxide production, leukotriene biosynthesis, and cytokine production. Each of these functional capabilities is linked to the production of tissue damage and physiologic derangements that are characteristic of human diseases associated with eosinophil-dominated inflammation, such as asthma. This review concerns the biology of eosinophils as it pertains to the pathogenesis of allergic disease. PMID- 9337425 TI - What exactly is exercise-induced asthma? PMID- 9337424 TI - In vivo and in vitro immunomodulation induced by the extract of the mycelium fungus Polyporus squamosus. AB - The mycelial mass of the fungus Polyporus Squamosus strain 64 (PS-64) was disintegrated by mechanical and ultrasound treatments. After centrifugation, the supernatant containing the disintegrate was dialyzed and lyophilized. The resultant PS-64 extract was subsequently investigated as an immunomodulator of IgE and IgG responses to ovalbumin (OA) in (CBAxC57BL/6)F1 mice using passive cutaneous anaphylaxis (PCA) and enzyme-linked immunosorbent assay (ELISA), respectively. Multiple injections of PS-64 extract in doses of 1.5, 15, and 150 mg/kg administered before the primary or secondary immunization of mice with OA resulted in a dose-dependent inhibition of both IgE and IgG antibody responses to OA. In contrast to the inhibition of the anti-OA IgE response noted during the entire 3-week observation period, the anti-OA IgG response was restored to control level by the third week of secondary immunization. The glass microfiber based whole blood histamine release assay demonstrated that various concentrations of the PS-64 extract did not influence histamine release induced either by anti-IgE or by specific allergens from basophils derived from whole blood of allergen-sensitized patients. Using the hemolytic plaque assay, significant suppression of IgM-secreting cell formation was noted in (CBAxC57BL/6)F1 mice administered various doses of the PS-64 extract before immunization. The PS-64 extract inhibited the in vitro proliferation of human mononuclear cells upon stimulation with phytohemagglutinin (PHA). In a dose dependent manner, the PS-64 extract also inhibited delayed-type hypersensitivity reaction and skin graft rejection, similar to the effect noted with usage of Cyclosporin A (CsA) in (CBAxC57BL/6)F1 mice. Our investigation suggests that the immunomodulatory effects of PS-64 should be studied further for potential clinical therapeutic utility. PMID- 9337427 TI - Nasal provocation tests in the diagnosis of urticaria induced by acetylsalicylic acid. AB - Nasal provocation tests with lysine acetylsalicylic acid (ASA) have been used in the diagnosis of ASA-induced asthma and rhinitis. To establish its possible role in identifying aspirin sensitivity manifested by urticaria or angioedema, 18 patients suffering from chronic or acute recurring urticaria/angioedema (10 ASA sensitive and 8 ASA-nonsensitive) were submitted to nasal provocation tests with freshly prepared solutions of lysine ASA. Clinical response and variation of nasal expiratory peak-flow were evaluated, classified according to previously defined scores, and compared. The results showed a significant difference between ASA-sensitive and ASA-nonsensitive patients, suggesting that this test can be an important diagnostic tool for ASA-induced urticaria/angioedema. PMID- 9337426 TI - Adverse reactions to acetaminophen, ASA, and NSAIDs in children: what alternatives? AB - ASA and NSAIDs are responsible for a large number of adverse reactions. The association of adverse reactions to acetaminophen and to ASA is uncommon, especially in children, and raises the problem of finding alternative treatments. We present a case report of a 7-year-old boy with combined adverse reaction to acetaminophen and ASA/NSAIDs. The child, who had no history of atopy, first displayed the condition at age 6, when he suffered two episodes of urticaria and angioedema, 2 hours after administration of 500 mg of acetaminophen, following two earlier doses of 500 mg (total 1500 mg). At age 7 he suffered a third episode 3 hours after administration of 180 mg of ASA. The patient submitted to oral challenges with acetaminophen (positive at a cumulative dose of 2,040 mg), ASA (positive at a cumulative dose of 204 mg) and nimesulide (negative at a cumulative dose of 119 mg). In conclusion, nimesulide (an NSAID not available in the United States) may be regarded as an alternative treatment in such patients, but more research is needed in pediatric age groups. PMID- 9337429 TI - Asthma among the famous. Edward VII (1841-1910) King of Great Britain. PMID- 9337428 TI - Allergic reaction to inadvertent peanut contact in a child. AB - Peanut anaphylaxis is a potentially near-fatal or fatal disease complicated by the fact that peanuts as well as other food items are commonly used as an adulterant in the preparation of foods. A boy is reported with peanut allergy to demonstrate, presumably for the first time, that contact urticaria occasionally provoked by peanuts can be associated with IgE-mediated allergy. Methods included skin prick tests, specific IgE determination, and open food challenge. All data were positive for an IgE-mediated allergy, and the open challenge with peanut resulted in systemic reactions. Food allergy is a common ailment in childhood. Although the ideal treatment is elimination of the offending allergen hidden, accidental, or unusual exposures can cause unwanted reactions, and anaphylaxis. The most reliable treatment appears to be prevention. PMID- 9337430 TI - Asthma among the famous. Ambrose Bierce (1842-1914), American journalist and author. PMID- 9337431 TI - Asthma among the famous. Marie-Bernarde Soubirous (1844-1879), French canonized Catholic nun: St. Bernadette. PMID- 9337432 TI - Asthma among the famous. Joseph Pulitzer (1847-1911), American journalist and publisher. PMID- 9337433 TI - The Jewish physician in the post Columbus era: the 15th century Spanish inquisition and expulsion revisited in 20th century Germany and Austria. PMID- 9337434 TI - Effect on dissolution from halving methylphenidate extended-release tablets. AB - OBJECTIVE: To determine the effect on in vitro dissolution from cutting methylphenidate extended-release tablets in half. DESIGN: Ritalin-SR (Ciba Pharmaceutical Co.) and generic methylphenidate extended-release (MD Pharmaceutical Inc.) tablets were dissolved in water according to the method prescribed by the US Pharmacopeia under two conditions: whole and halved. Samples were collected at 15, 30, and 45 minutes and at 1, 2, 3, 3.5, 4, 5, 6, and 7 hours. Methylphenidate content was determined by HPLC. RESULTS: Halving the tablets caused a statistically significant increase in cumulative dissolution as early as 15 minutes. The difference in cumulative dissolution reached its maximum for both Ritalin-SR and generic methylphenidate extended-release tablets at 2 hours. At this time point, the percent dissolution of the whole versus halved tablets was 57% versus 74% (Ritalin-SR), respectively, and 49% versus 67% (generic), respectively. The dissolution profiles of halved and whole extended release methylphenidate tablets were parallel from this point through the 7-hour collection period. At 7 hours, however, there was no difference in the cumulative dissolution of halved versus whole tablets. CONCLUSIONS: While statistical differences during in vitro dissolution do exist and pharmacokinetic ramifications have not yet been determined, the absolute differences in dissolution between halved and whole tablets are not great. Halving methylphenidate extended-release tablets may be a clinically acceptable means of achieving a small increment/decrement in dose without converting to a regular release tablet. PMID- 9337435 TI - Gentamicin and tobramycin pharmacokinetics in pediatric bone marrow transplant patients. AB - OBJECTIVE: To describe the pharmacokinetic parameters of gentamicin and tobramycin in pediatric bone marrow transplant patients. DESIGN: Retrospective medical record review. SETTING: Pediatric bone marrow transplant unit in a university teaching hospital. MAIN OUTCOME MEASURES: Pharmacokinetic parameters (apparent volume of distribution [Vd] in L/kg, half-life [t1/2] in h, elimination rate constant [ke] in h-1, clearance [Cl] in mL/min/1.73 m2 and mL/min/kg) calculated from serum concentrations. PATIENTS: Thirty-three patients aged 15 years or less who underwent bone marrow transplant and received gentamicin or tobramycin. RESULTS: Mean pharmacokinetic parameters were Vd 0.32 +/- 0.07 L/kg, t1/2 2.32 +/- 0.65 h, Cl 1.71 +/- 0.53 mL/min/kg, and Cl 86.2 +/- 24.5 mL/min/1.73 m2. Factors such as disease state, type of marrow graft, gender, or exposure to cyclosporine had no significant effect on pharmacokinetic parameters. Linear regression indicated a weak relationship between serum creatinine (SCr) and Cl in mL/min/kg (r = 0.59), but no relationship was found between SCr and Cl in mL/min/1.73 m2, between age and apparent Vd, or between SCr and apparent Vd. Models for estimating Cl and Ke developed by multiple regression were somewhat predictive (r = 0.7). Required calculated maintenance dosages to obtain therapeutic concentrations were 8, 7, and 6 mg/kg/d in children 6 or younger, 7 12, and 13-15 years, respectively. CONCLUSIONS: The mean Cl and apparent Vd for all ages are similar to those reported in pediatric oncology patients who had not undergone marrow transplantation. Children 6 years or younger had lower than expected Cls and larger apparent Vds than did the older children. Dosages estimated to be necessary to achieve therapeutic concentrations were 6-8 mg/kg/d. PMID- 9337436 TI - Removal of vancomycin during plasmapheresis. AB - OBJECTIVE: To examine the removal of vancomycin during plasmapheresis, determine whether drug administration should be withheld prior to or a supplemental dose given after the procedure, and determine whether a redistribution phenomenon in vancomycin serum concentrations occurs after plasmapheresis. DESIGN: Prospective, cohort study. SETTING: An 800-bed, tertiary-care, teaching hospital. PATIENTS: Twelve patients receiving vancomycin as prescribed who were also undergoing therapeutic plasmapheresis. METHODS: Blood samples for determination of vancomycin concentrations were obtained from each patient immediately before, during, immediately after, and 2 hours after plasmapheresis. Vancomycin concentration in plasma removed by plasmapheresis and volume of plasma removed were measured. Patient-specific pharmacokinetic parameters were determined for each patient using serum concentration data and a one-compartment model. Percent of drug removed by plasmapheresis and percent increase in vancomycin total clearance secondary to plasmapheresis were calculated. RESULTS: A mean of 6.3% of the total body store of vancomycin was removed by plasmapheresis. Vancomycin clearance during plasmapheresis averaged 1.6 L/h, which was an average increase of 285% in the total clearance of vancomycin from the body. Nine of 10 patients had a higher observed vancomycin concentration 2 hours after plasmapheresis than that predicted by degrading the concentration observed immediately after the procedure, suggesting that redistribution in serum concentrations occurs after the procedure. CONCLUSIONS: A single one-volume plasmapheresis does not remove a clinically important amount of vancomycin; therefore, supplemental dosing after the procedure is not necessary. A redistribution phenomenon in vancomycin concentrations appears to exist after plasmapheresis. Further study is needed to determine how long the redistribution phase lasts and when vancomycin concentrations should be measured after plasmapheresis. PMID- 9337437 TI - Cost-effectiveness comparison of sequential ofloxacin versus standard switch therapy. AB - OBJECTIVE: To compare the cost-effectiveness of sequential intravenous-to-oral ofloxacin versus intravenous-to-oral standard switch therapy for the treatment of patients with sepsis who are hospitalized with bacterial infections. DESIGN: Cost effectiveness analysis from a provider perspective, including resources important to an integrated healthcare network, of a randomized, open-label, controlled, clinical trial. SETTING: Millard Fillmore Health System, Buffalo, NY. PATIENTS: Hospitalized adults requiring parenteral antibiotics for a complicated urinary tract infection, lower respiratory tract infection, or skin and soft tissue infection. INTERVENTIONS: Sequential intravenous-to-oral ofloxacin or standard intravenous-to-oral switch antibiotics. OUTCOME MEASURES: Clinical outcomes and direct costs associated with hospitalization, primary physician services, specialist physician services, and outpatient care. RESULTS: Eighty-two of 89 patients randomized into the two treatment groups were evaluable. Standard switch therapy failed with 12 patients versus 10 patients receiving ofloxacin. Complete economic data were available for 74 patients. Sequential ofloxacin therapy resulted in a 1-day-shorter antibiotic-related hospitalization without evidence of recurrent infection during the posttherapy follow-up evaluations. An average cost savings of $399 per patient was achieved in the sequential ofloxacin group. Although this difference did not attain statistical significance (probably due to the large variance), it is an economically significant finding. The cost effectiveness ratios were $5735 per successful outcome for the standard switch therapy group versus $5126 per successful outcome in the sequential ofloxacin group. CONCLUSIONS: Sequential ofloxacin was as effective and consistently less expensive than standard switch antibiotics in the initial evaluation and in the sensitivity analysis of room cost and drug acquisition cost. Standard switch therapy would have to be greater than 25% more effective than sequential ofloxacin therapy to change the economic decision. PMID- 9337438 TI - Indinavir sulfate renal toxicity in a pediatric hemophiliac with HIV infection. AB - OBJECTIVE: To report a case of renal toxicity associated with administration of indinavir sulfate in a pediatric hemophiliac with HIV infection. CASE SUMMARY: A 16-year-old white hemophiliac boy with HIV infection secondary to tainted coagulation factor VIII was treated with indinavir sulfate. The patient developed gross hematuria, proteinuria, pyuria, abdominal pain, increased bilirubin, an elevated serum creatinine (SCr) of 1.2 mg/dL (baseline 0.9-1.0), and symptoms of renal colic within 1 month of starting indinavir sulfate therapy. Approximately 2 months later the patient developed a low-grade fever with a further increase in SCr. He was prescribed a 10-day course of cefpodoxime proxetil for a possible urinary tract infection. One week later, the patient developed fever, chills, nausea, vomiting, decreased appetite, sterile pyuria, nasal congestion, and an elevated SCr of 1.3-1.7 mg/dL. Indinavir sulfate and cefpodoxime proxetil were discontinued and the patient was suspected of having tubulointerstitial nephritis secondary to indinavir sulfate. The patient's nephritis resolved and the SCr decreased to 1.1 mg/dL within 1 month of discontinuing indinavir sulfate. CONCLUSIONS: This case demonstrates the potential for renal toxicity with the use of indinavir sulfate in HIV-infected hemophiliacs. PMID- 9337439 TI - Debilitating reaction following the initial dose of tramadol. AB - OBJECTIVE: To describe a debilitating reaction following a single oral dose of tramadol. CASE SUMMARY: A 32-year-old white man with no prior medical problems, allergies, or previous medication reactions experienced ataxia, dilation of the pupils, numbness in his arms and legs, tremulousness, and dysphoria lasting approximately 4 hours following an initial tramadol dose (100 mg). The patient recovered with no sequelae. DISCUSSION: Central nervous system (CNS) stimulation during therapy with tramadol was reported in 7% of patients in clinical trials. These reactions are usually mild and transient. This report describes a debilitating CNS-mediated reaction to an initial dose of tramadol in an otherwise healthy adult. The patient was phenotyped for CYP2D6 activity, the major metabolic pathway for tramadol elimination, and was determined to be an extensive metabolizer with very high CYP2D6 activity. CONCLUSIONS: The exact mechanism of the adverse response is not known; however, based on phenotyping results, we suspect that it may be related to high concentrations of the active O-desmethyl metabolite of tramadol. PMID- 9337440 TI - Pharmacokinetics of methotrexate in plasma and cerebrospinal fluid. AB - OBJECTIVE: To investigate the pharmacokinetics of methotrexate (MTX) in plasma and cerebrospinal fluid (CSF) during osmotic disruption of the blood-brain barrier and the intraarterial administration of combination chemotherapy postoperatively in a patient with glioblastoma. CASE SUMMARY: A 60-year-old Japanese woman with a glioblastoma received two courses of combination intraarterial chemotherapy. In the first course of treatment, 20 mL of mannitol 20%, peplomycin 10 mg, vindesine 2 mg, and MTX 500 mg were administered via the right internal carotid artery, and then via the right vertebral artery. In the second course of treatment, 20 mL of mannitol 20%, peplomycin 15 mg, vindesine 2.5 mg, and MTX 1000 mg were similarly administered. Blood samples and CSF samples from the ventricle and the space left by tumor removal were obtained; the MTX concentrations were measured from these sites by fluorescence polarization immunoassay. The pharmacokinetic parameters of MTX in plasma and CSF were estimated. DISCUSSION: The plasma concentration of MTX decreased in a biexponential decay pattern during each course of treatment. CSF concentrations of MTX in the ventricle and in the space left by tumor removal peaked at 2 and 6 hours, respectively, after drug administration and decreased monoexponentially. When the dose of MTX was doubled, the AUC for the plasma MTX concentration increased 2.4-fold and the AUCs for MTX in the ventricle and the space left by tumor removal increased 3.4- and 9.1-fold, respectively. The half-life of MTX in the CSF in the space left by tumor removal exceeded the half-lives of MTX in the plasma and in the ventricular CSF. CONCLUSIONS: The CSF AUCs of MTX in the ventricle and the space left by tumor removal increased markedly and in parallel with the MTX dosage increase during osmotic disruption of the blood-brain barrier and intraarterial combination chemotherapy. Such treatment improves the delivery of chemotherapy agents to the brain. PMID- 9337441 TI - Phenolphthalein-induced toxic epidermal necrolysis. AB - OBJECTIVE: To report a case of phenolphthalein-induced toxic epidermal necrolysis (TEN) in a patient maintained on several other medications more commonly known to be associated with TEN. CASE SUMMARY: A 78-year-old white man presented with intractable lower back pain and constipation. On day 1 of admission, the patient exhibited a diffuse urticarial rash over his trunk and extremities. History revealed that the patient had taken a combination phenolphthalein/docusate sodium (Correctol) over-the-counter laxative 1 day prior to admission. He had a similar urticarial rash 1.5 years earlier with this product and was instructed not to use it. A biopsy was performed and evidence from light microscopic analysis of the tissue led to a diagnosis of TEN. Furosemide, spironolactone, allopurinol, and hydroxyurea were considered possible causes of the reaction and were discontinued. Despite this, the lesions worsened in severity. The patient subsequently responded well to intravenous antibiotics, intravenous corticosteroids, and local wound care. Furosemide, spironolactone, hydroxyurea, allopurinol, and docusate were all reintroduced without reactivation of the lesions. DISCUSSION: Phenolphthalein is the active ingredient in several over-the counter laxative preparations and has only rarely been reported to cause TEN. (It is no longer contained in Correctol.) To our knowledge, this case report represents only the third description of laxative-induced TEN. Although this patient had been exposed to several other medications more commonly associated with TEN, his long-term tolerance of and uneventful rechallenge with these medications exclude them as potential catalysts to this drug reaction. The patient's previous rash and the temporal relation of this event and the ingestion of phenolphthalein, as well as the similarity of this case to other reports, point to phenolphthalein as the cause of TEN in this patient. CONCLUSIONS: TEN is a rare disorder that can be fatal in up to 30% of patients. Clinicians should include phenolphthalein in their list of possible causes of drug-induced TEN. A careful and complete medication history can help avoid unnecessary discontinuation of clinically important medications and inadvertent rechallenge with the causative agent. PMID- 9337442 TI - Minocycline-induced lupus. AB - OBJECTIVE: To report a case of drug-induced lupus occurring 5 months after the initiation of minocycline therapy for acne. DATA SOURCE: Case report information was obtained from the physician, patient's family, and the medical record. MEDLINE and Index Medicus were searched to obtain relevant published literature from 1966 to 1996. CASE SUMMARY: A 14-year-old white girl developed symptoms of myalgias, arthralgias, polyarthritis, and flushed face. The antinuclear antibody test was positive. Minocycline was discontinued and the patient's condition dramatically improved within 7 days. CONCLUSIONS: Healthcare providers should recognize early common and unusual symptoms of minocycline-induced lupus in adolescents being treated for acne. PMID- 9337443 TI - Topiramate: a new antiepileptic drug. AB - OBJECTIVE: To review proposed mechanisms of action, clinical pharmacology, efficacy, safety, and tolerability of the antiepileptic drug (AED) topiramate. METHODS: All available data from the clinical development program--published and unpublished data (provided by investigators or the RW Johnson Pharmaceutical Research Institute)--were analyzed, with emphasis on the results of double-blind, placebo-controlled trials. Data from open-label studies provided information about long-term efficacy and tolerability. FINDINGS: Topiramate is highly effective in the control of previously therapy-resistant partial seizures, with or without secondary generalization. In the refractory adult patient population enrolled in controlled clinical trials, seizures were reduced by 50% or more in 27-47% of patients compared with 0-18% in placebo-treated patients and by 75% or more in 9-36% of the patients compared with 0-9% of those receiving placebo. The most common adverse effects involve the central nervous system and are mild to moderate in nature. Adverse effects include somnolence, fatigue, psychomotor slowing, and concentration problems. The currently recommended dosing is lower and titration slower than schedules used in the clinical trials; this may improve the tolerability of topiramate. Topiramate has few interactions with currently available AEDs, but phenytoin concentrations increased in some patients. Topiramate can be used initially as adjunctive therapy. Plasma topiramate concentrations are reduced substantially when used with enzyme-inducing AEDs. Open-label data and a single double-blind trial suggest that topiramate monotherapy may be effective. Open-label data also indicate that topiramate may be effective in generalized seizures of nonfocal origin, including those associated with Lennox-Gastaut syndrome. CONCLUSIONS: The robust clinical effects of topiramate expand the therapeutic options for patients with epilepsy. Controlled clinical trials are needed to verify initial observations that topiramate may be effective against a broad spectrum of seizure types and to directly compare efficacy and tolerability with other new and standard AEDs. PMID- 9337445 TI - Primary stroke prevention in nonvalvular atrial fibrillation: implementing the clinical trial findings. AB - OBJECTIVE: To review the clinical trials evaluating warfarin for primary stroke prophylaxis in nonvalvular atrial fibrillation (NVAF), to discuss the relative benefits and risks of warfarin versus aspirin therapy, and to review the clinical practice guidelines and identify potential barriers to their implementation in clinical practice. DATA SOURCES: A MEDLINE literature search was performed to identify clinical trials of antithrombotic therapy for NVAF, clinical practice guidelines, studies evaluating physician practices and attitudes, cost effectiveness studies, and pertinent review articles. Key search terms included atrial fibrillation, stroke, antithrombotic, warfarin, aspirin, and cost effectiveness. DATA EXTRACTION: Prospective, randomized clinical trials were selected for analysis. Clinical practice guidelines from recognized panels of experts were reviewed. Comprehensive review articles were selected. DATA SYNTHESIS: NVAF is a common arrhythmia that is associated with a substantial risk for stroke. Seven prospective, randomized, clinical trials have conclusively demonstrated the efficacy of warfarin for stroke prevention. The greatest benefits are achieved in older patients and those with comorbidities that increase their risk for stroke. The potential benefits of preventing a devastating stroke, however, must be weighed against the potential for bleeding complications. Warfarin has been shown to be cost-effective in high-risk patients, provided the rate of complications is minimized. Nonetheless, many physicians remain hesitant to implement warfarin therapy in older, high-risk patients. The clinical data on aspirin are less consistent than those observed with warfarin. Aspirin appears to be most effective in younger individuals or those considered to be at low risk for stroke. CONCLUSIONS: In patients with NVAF, the personal, social, and economic consequences of stroke are often devastating. Clinical trials have provided definitive proof that the risks of stroke can be significantly reduced through the use of appropriate antithrombotic therapy. Despite this evidence and the recommendations of a number of clinical practice guidelines, variations in care exist that continue to place patients at risk. Additional outcomes research is needed to evaluate the impact of the clinical trial findings and practice guidelines on clinical practice and to develop methods for overcoming barriers to implementation. PMID- 9337444 TI - Amphotericin B lipid complex. AB - OBJECTIVE: To evaluate the published data on the effectiveness and safety of amphotericin B lipid complex (ABLC) for the treatment of invasive mycosis and to evaluate data describing the pharmacologic properties and pharmacokinetic behavior of ABLC in both animals and humans. DATA SOURCE: A MEDLINE search was conducted to identify literature published from 1965 to January 1997 for amphotericin B deoxycholate (DCAB) and ABLC. In addition, preliminary data published as abstracts and presented at national conferences on infectious disease and hematology within the last 6 years were also included in this review. STUDY SELECTION: Both human and animal studies were reviewed. Animal and in vitro studies were selected to evaluate the pharmacologic and toxicologic properties of ABLC. For the evaluation of the efficacy, safety, and pharmacokinetic behavior of ABLC, large, well-controlled studies were reviewed. In addition, data from open label and emergency use protocols were also included in the review. DATA EXTRACTION: The study and analytical methods, results, and conclusions of the selected studies were evaluated. Pharmacokinetic data for both ABLC and DCAB that were derived from human subjects were also evaluated. DATA SYNTHESIS: DCAB has been the cornerstone for the treatment of invasive mycosis, even though it has a narrow therapeutic index. Infusion-related toxicities (e.g., fever, chills, rigors) are likely due to DCAB stimulation of cytokine and prostaglandin synthesis. Conversely, nephrotoxicity, the primary non-infusion-related toxicity, likely results from the nonselective cytotoxic interaction between DCAB and cholesterol-containing mammalian cells. ABLC represents a new approach to improving the therapeutic index of DCAB. Mammalian cytotoxicity is attenuated by complexing amphotericin B to a mixture of phospholipids. This alters the affinity of amphotericin B and decreases its selective transfer from the complex to cholesterol-containing mammalian cells. Fungi also possess lipase, which improves the selective transfer from the complex to ergosterol-containing cell membranes. In humans, the lipid formulation increases the volume of distribution of amphotericin B. Thus, compared with DCAB, larger doses of ABLC can be administered for a longer duration with less nephrotoxicity. However, the prevalence of infusion-related toxicities associated with ABLC is similar to that of DCAB. Whether the alteration in distribution improves efficacy by improving tissue concentrations of amphotericin B has not been determined. The cost of this agent will limit its use. CONCLUSIONS: ABLC has been shown to be at least as effective as DCAB, and it has been well tolerated in the clinical studies to date. Despite large dosages and extended courses of administration, there is little nephrotoxicity associated with its use. However, the cost of this agent will limit its use to the treatment of refractory mycosis or to cases where DCAB is contraindicated due to significant renal insufficiency. PMID- 9337446 TI - The AHCPR clinical practice guideline for heart failure revisited. AB - OBJECTIVE: To review the Agency for Health Care Policy and Research (AHCPR) clinical practice guideline for heart failure and comment on the guideline regarding pharmacotherapy from the perspective of the latest clinical trial data and the authors' clinical experience. DATA SOURCES: A MEDLINE search (1966 to June 1997) of English-language literature pertaining to the pharmacotherapy of heart failure was performed. Special emphasis was placed on literature published in the last 5 years. Additional literature was obtained from reference lists of key articles identified through the search. DATA SYNTHESIS: Pertinent clinical trials were reviewed and considered along with information from the authors' database of over 800 patients with heart failure. Evidence concerning the use of angiotensin-converting enzyme inhibitors at appropriate dosages in all New York Heart Association classes of heart failure and the inclusion of digoxin as part of triple therapy in all symptomatic patients with left ventricular systolic dysfunction are reviewed. Strategies to circumvent clinical problems that may limit the proper application of standard therapeutic agents are considered, and the possible future role of beta-blockers as the therapeutic agents in patients with heart failure is discussed. CONCLUSIONS: The AHCPR guideline provides the clinician with an excellent framework for treating the patient with heart failure. Building on the fundamentals of the guideline, the clinician can carefully apply current therapy at appropriate dosages and in the best combinations to individualize and thereby optimize pharmacologic therapy for this patient population. PMID- 9337447 TI - New pharmacotherapy for Parkinson's disease. AB - OBJECTIVE: To summarize the development, pharmacology, pharmacokinetics, efficacy, and safety of five investigational antiparkinsonian drugs that are in or have recently completed Phase III trials: three dopamine agonists, pramipexole, ropinirole, and cabergoline; and two catechol-O-methyltransferase (COMT) inhibitors, entacapone and tolcapone. The pathophysiology and the role of dopamine in Parkinson's disease are also reviewed. DATA SOURCES: A MEDLINE search of relevant English-language literature, clinical studies, abstracts, and review articles pertaining to Parkinson's disease was conducted. Manual searches of 1996/1997 meeting abstracts published by the American Academy of Neurology and the Movement Disorders Society were also performed. Manufacturers provided unpublished Phase III trial efficacy and pharmacokinetic data. STUDY SELECTION AND DATA EXTRACTION: Clinical trial investigations selected for inclusion were limited to human subjects. Interim analyses after 6 months for long-term clinical studies in progress were included. Pharmacokinetic data from animals were cited if human data were unavailable. Statistical analyses for all studies were evaluated. DATA SYNTHESIS: By selectivity targeting D2 receptors, the newer dopamine agonists (i.e., cabergoline, pramipexole, ropinirole) may delay the introduction of levodopa and thus the occurrence of levodopa-induced dyskinesias. In addition, they are efficacious as adjunctive therapies in patients with advanced Parkinson's disease. Unlike the currently available dopamine agonists, pramipexole and ropinirole are non-ergot derivatives and do not cause skin inflammation, paresthesias, pulmonary infiltrates, or pleural effusion. The COMT inhibitors, tolcapone and entacapone, improve the pharmacokinetics of levodopa by preventing its peripheral catabolism and increasing the concentration of brain dopamine; thus, these agents may reduce the incidence of "wearing-off" effects associated with the short half-life of levodopa and the progression of Parkinson's disease. CONCLUSIONS: Interim 6-month analyses of pramipexole, ropinirole, and cabergoline for symptomatic treatment of early Parkinson's disease have shown these drugs to be efficacious and relatively well-tolerated when used as monotherapy. Their role in delaying the development of motor fluctuations and delaying the addition of levodopa is the subject of long-term clinical studies. In advanced stages of Parkinson's disease, these medications were also efficacious; however, the main adverse effects included dyskinesias, somnolence, and hallucinations. The COMT inhibitors, entacapone and tolcapone, have also demonstrated efficacy in improving on-time in patients with stable disease. Tolcapone has also demonstrated efficacy in patients with motor fluctuations. Both drugs are relatively well-tolerated, with the exception of dyskinesias that require reduction of the levodopa dosage and occasional diarrhea. PMID- 9337448 TI - Melatonin: therapeutic use in sleep disorders. AB - OBJECTIVE: To review the use of melatonin in sleep disorders, including jet lag, shift work disorder, insomnia, and sleep cycle disorders in neurologically impaired patients. DATA SOURCE: A MEDLINE search (1966 to April 1996) was performed that included clinical studies and reviews on melatonin in the English language. References used in those articles were also screened for additional information. STUDY SELECTION: All published trials were considered for inclusion in this review, with an emphasis placed on more recently published studies (last 5 years). DATA SYNTHESIS: There is significant evidence that links the hormone melatonin to circadian sleep cycles in humans. It has been suggested that in situations where the endogenous melatonin concentration is reduced (advancing age) or the normal circadian cycle is disrupted (jet lag, shift work, blind patients), supplementation with exogenous melatonin may improve both sleep duration and quality. Limited data from generally short-term trials and anecdotal reports suggest that melatonin may be effective in several of these sleep disorders. Melatonin use in jet lag appears to decrease jet lag symptoms and hasten the return to normal energy levels. Melatonin may be helpful in rotating shift schedules to improve sleep quality and maintain normal circadian rhythm. In some patients with insomnia, melatonin appears to induce sleep onset. The optimal dosage and timing of drug administration is still unclear. CONCLUSIONS: Although there is some evidence that melatonin may have modest efficacy, especially in insomnia, jet lag, and sleep disorders in neurologically impaired patients, adequate long-term studies examining both efficacy and toxicity are lacking. In addition, further studies evaluating dose-response relationships and drug interactions are warranted. PMID- 9337449 TI - Pharmacologic management of supraventricular tachycardias in children. Part 1: Wolff-Parkinson-White and atrioventricular nodal reentry. AB - OBJECTIVE: To review the literature regarding the use of antiarrhythmic agents in the management of Wolff-Parkinson-White (WPW) syndrome and atrioventricular nodal reentry tachycardia (AVNRT) in infants and children, and to discuss the advantages and disadvantages of specific agents in each arrhythmia in an effort to develop treatment guidelines. DATA SOURCES: A MEDLINE search encompassing the years 1966-1996 was used to identify pertinent literature for discussion. Additional references were found in the articles that were retrieved via MEDLINE. STUDY SELECTION: Clinical trials that address the use of antiarrhythmic agents for the treatment of the supraventricular tachycardias WPW and AVNRT in children were selected. Literature pertaining to dosage, pharmacokinetics, efficacy and toxicity of antiarrhythmic agents in children were considered for possible inclusion in the review, and information judged to be pertinent by the authors was included in the discussion. DATA EXTRACTION: Although there are numerous reports of antiarrhythmic use in children, very few large studies are designed to evaluate an individual antiarrhythmic agent for a specific arrhythmia. Controlled, comparison trials of antiarrhythmic agents in children are virtually nonexistent. Ideally, controlled clinical trials are used to develop clinical guidelines; however, in this situation, most data and information must be obtained from case series of children treated. Although the results from these type of studies may be useful in developing guidelines for the optimal use of these agents, controlled trials are required for establishing standard treatment guidelines for all patients. DATA SYNTHESIS: Despite limited scientific evaluation of conventional agents in the treatment of WPW and AVNRT in children, they continue to be used as standard of care. Most information regarding the use of conventional agents in children has been extrapolated from the adult literature. Little justification for the use of agents or dosing in children is available. Controlled trials regarding the use of new antiarrhythmic agents (propafenone, amiodarone, flecainide) are available; however, the variance in dosing schemes, presence of structural heart disease, and patient age make the development of recommendations difficult. CONCLUSIONS: Because of greater clinical experience with these conventional antiarrhythmic agents, they continue to be first-line therapy in the management of most supraventricular tachycardia (SVT) in children. The management of SVT in children with WPW syndrome should begin with the use of a beta-blocker with the addition of digoxin or procainamide for treatment failures. The use of digoxin monotherapy, although frequently used by many practitioners in infants and children with WPW, cannot be recommended. For failures to conventional agents, flecainide is the preferred agent, while therapy with propafenone, amiodarone, and sotalol remains to be elucidated. The management of AVRNT is similar to that of WPW; however, digoxin is the agent of first choice. Trials of beta-blockers and procainamide should follow for treatment failures with flecainide again being the preferred "newer" antiarrhythmic for use in resistant cases. Additional well-designed, controlled trials are needed to further evaluate the comparative efficacy of antiarrhythmics in the management of WPW and AVNRT in children, as well as to evaluate dosing and toxicity in various age groups. PMID- 9337450 TI - Adjunctive intracoronary thrombolysis in complicated coronary angioplasty. AB - Percutaneous transluminal coronary angioplasty is complicated by abrupt closure in 4.4-9.5% of procedures. Although the etiology of closure is difficult to determine, arterial dissection and thrombus formation are often involved. When abrupt closure occurs, repeat balloon dilation of the affected vessel is the mainstay of treatment and results in a mean angiographic success rate of 44% (range 35-51%). Other interventions, such as stent implantation and atherectomy, may also be attempted. I.c. thrombolysis is an alternative rescue strategy for the treatment of abrupt coronary closure during angioplasty. Initial angiographic success with i.c. thrombolysis, in combination with repeat balloon dilation ranges from 52% to 90%. These results are encouraging, but vessel reocclusion occurs in up to 55% of patients, resulting in diminished clinical success. Two trials suggest thrombolysis is ineffective or detrimental in this patient population. Most studies evaluating i.c. thrombolysis are retrospective, noncomparative, lack standardized protocols, and evaluate dissimilar patient populations. Therefore, the contribution of confounding variable such as operator experience, balloon size, duration of balloon inflation, and investigator bias cannot be assessed. I.c. thrombolysis has a limited role in the treatment of abrupt closure. This therapy should be considered only if thrombus formation is definitively the cause of occlusion, and avoided if intimal dissection is present, due to possible detrimental effects. The results of thrombolysis as a sole rescue therapy for abrupt closure are disappointing. Therefore, repeat balloon dilation should always be performed concomitantly with drug administration. In select patients, streptokinase, alteplase, or urokinase may be given for abrupt closure. Urokinase is favored due to increased experience with this agent and decreased cost. Ambrose recommends 250,000-1,000,000 units of urokinase, infused for up to 30 minutes (average wholesale price $419-1676). Additional data indicate a lower dose of urokinase may be sufficient for closure resolution, but this has not been adequately assessed. I.c. rather than intravenous thrombolytic administration may cause fewer systemic effects; however, contraindications to thrombolytic therapy should always be evaluated and weighed against potential benefits. The future role of thrombolysis in the treatment of complicated coronary angioplasty is unclear. Only randomized, controlled trials can evaluate the merits of this treatment approach compared with other rescue strategies. PMID- 9337451 TI - Guidelines for the treatment of Helicobacter pylori in the pediatric population. AB - Several factors including long-term eradication of the organism, cost, compliance, and adverse event profile should be considered for treating H. pylori infection in pediatric patients. Triple therapy with bismuth, tetracycline, and metronidazole is considered the gold standard for adult patients; however, tetracyclines are not recommended in children younger than 8 years due to the potential for tooth discoloration and alterations in bone growth. Dual and shorter duration of therapy should be evaluated in children with H. pylori. The new dual therapy omeprazole/clarithromycin regimens approved by the Food and Drug Administration for adults may be considered as an alternative for children when concerns include the use of salicylates or allergy to beta-lactams. Although the dosage of omeprazole in pediatric patients has not been established (no pediatric formulation exists), clarithromycin is available for use in pediatric patients. However, these drugs cannot be recommended for children with H. pylori until additional studies in this population are available. Based on the available data, aminopenicillin/bismuth or aminopenicillin/tinidazole combinations appear to be effective in eradicating H. pylori in children. Amoxicillin 50 mg/kg/d plus bismuth subsalicylate (< 10 y, 262 mg; > 10 y, 525 mg qid) or bismuth subcitrate (< 12 y, 120 mg; > 12 y, 240 mg bid) can be used for 6 weeks. The bismuth dosages represented above were those used in various studies. It should be realized, however, that a definitive dosage of bismuth subsalicylate for children in the treatment of H. pylori has not been established. The adult dosage of bismuth subsalicylate for the eradication of H. pylori is the same as that used for prophylaxis in diarrhea (525 mg qid). When dosage of this agent is unknown (particularly for the treatment of very young children), the use of established dosages for prophylaxis in diarrhea may be considered for treating H. pylori. Additionally, bismuth subsalicylate should be used with caution in children with suspected viral infections (i.e., to prevent Reye's syndrome) or those receiving concurrent therapy with interacting drugs. If available, tinidazole 20 mg/kg/d can be used with amoxicillin 50 mg/kg/d for 6 weeks to treat children infected with H. pylori. PMID- 9337452 TI - Ursodiol to prevent hepatic venoocclusive disease. AB - VOD is an important cause of morbidity and mortality in patients following allogeneic bone marrow transplantation. Although VOD may improve in some patients, severe cases are often fatal. There is no established therapy to prevent progression to severe VOD; nor are there any conclusive or universally accepted methods for prevention of mortality associated with severe VOD. A treatment that could minimize hepatic damage and cause VOD to manifest as reversible liver damage rather than a progressive, fatal disease would indeed have a place in posttransplant therapy. Nontoxic ursodiol may play such a role by replacing hepatotoxic bile acids. Based on the limited available literature (2 studies), it is difficult to draw firm conclusions regarding the use of ursodiol to prevent VOD, although the preliminary results are promising. The studies, although small and not without weakness, suggested that ursodiol effectively reduces the incidence of VOD in allogeneic BMT patients. They do not, however, suggest that ursodiol is effective as treatment for existing VOD, as this was not studied. Also, conclusions were based on patients given busulfan and cyclophosphamide as conditioning therapy and thus might not apply to patients conditioned by other means such as total body irradiation. In summary, the available data do not definitively support the use of ursodiol; however, patients conditioned with busulfan and cyclophosphamide who are at risk for VOD (e.g., pretransplant liver disease, liver metastases) may be candidates for ursodiol prophylactic therapy. Larger, randomized clinical trials are warranted to further define the potential role of ursodiol for the prevention of venoocclusive disease of the liver in BMT patients. PMID- 9337453 TI - Pharmacy and tobacco. AB - With the recognition that smoking begins in youth and that tobacco products are readily available to those under 18 years of age, new Food and Drug Administration (FDA) regulations restrict the sale, distribution, promotion, and advertising of cigarettes to minors. The objective is to decrease the use of tobacco by young people and consequently reduce the future morbidity and mortality from tobacco. Pharmacists currently have three choices with regard to the sale of tobacco in pharmacies: display and sell tobacco products, refuse to sell tobacco products, or make tobacco products available but counsel on smoking cessation. Each choice, as well as the impact of the new FDA regulations on pharmacy, is discussed. PMID- 9337454 TI - Trimethoprim/sulfamethoxazole-induced hypersensitivity syndrome. PMID- 9337455 TI - Alopecia associated with gabapentin: first case. PMID- 9337456 TI - Anticholinergic delirium possibly associated with protriptyline and fluoxetine. PMID- 9337457 TI - Delayed elimination of methotrexate associated with piperacillin administration. PMID- 9337458 TI - Dissolution of acetaminophen tablets under overdose conditions. PMID- 9337459 TI - Improvements in drug information for patients with special needs. PMID- 9337460 TI - Hiccups following nicotine gum use. PMID- 9337461 TI - Comment: drug-induced hiccups. PMID- 9337463 TI - Glossary of times used for scheduling and monitoring of diagnostic and therapeutic procedures. PMID- 9337462 TI - Surgical hand scrub time. PMID- 9337464 TI - Management of sports-related anterior cruciate ligament injuries. AB - Injuries from recreational and competitive sports activities have increased significantly during the past two decades. The anterior cruciate ligament (ACL) is one of the most commonly injured knee ligament in athletes. As the ACL is the knee's primary intra-articular stabilizer, injuries to this ligament can seriously alter athletes' physical activity levels or end their sports careers. The medical and surgical treatment options for a sports-related ACL injury depend upon the age of the patient, the type and degree of the ACL injury, and the anticipated future activity level of the athlete. This article describes the anatomy and physiology of the knee joint, the different types and degrees of ACL injuries, diagnostic measures for ACL injuries, and treatment options, which include arthroscopically assisted ACL reconstruction procedures with autologous patellar tendon grafts. Preoperative, intraoperative, and postoperative nursing interventions, rehabilitation regimens, and sports injury prevention measures are discussed. PMID- 9337465 TI - Laparoscopic spinal fusion procedures. AB - Laparoscopic procedures, such as spinal fusions, facilitate patients' faster returns to functioning lifestyles. Fusions using implanted threaded cylinders made of a porous titanium alloy can be performed at the cervical, thoracic, and lumbar spine levels. This article discusses preoperative, intraoperative, and postoperative care of patients undergoing laparoscopic spinal fusion. PMID- 9337466 TI - Guided imagery as a coping strategy for perioperative patients. AB - Patients who undergo surgery usually experience fear and apprehension about their surgical procedures. Guided imagery is a simple, low-cost therapeutic tool that can help counteract surgical patients' fear and anxiety. The authors randomly assigned 130 patients undergoing elective colorectal surgical procedures into two groups. Members of one group received routine perioperative care. Members of the other group listened to guided imagery tapes for three days before their surgical procedures, during anesthesia induction, intraoperatively, in the postanesthesia care unit, and for six days after surgery. The authors measured patients' anxiety levels, pain perceptions, and narcotic medication requirements. The patients in the guided imagery group experienced considerably less preoperative and postoperative anxiety and pain, and they required almost 50% less narcotic medications after their surgical procedures than patients in the control group. PMID- 9337467 TI - Competency validation for cross-training in surgical services. AB - Cross-training is a cost-effective means of maximizing the potential of staff members and available resources. The responsibility to provide quality patient care should not be compromised by cross-training efforts. Surgical services department staff members at Deaconess-Nashoba Hospital, Ayer, Mass, developed a competency validation project for nurses involved in the cross-training program. They accomplished this by implementing current nursing standards and practices and promoting continuity of patient care through the preoperative, intraoperative, and postoperative phases. Perioperative nurses and health care facilities have the responsibility to provide safe, effective, and efficient care to all patients. By assessing competencies for perioperative nurses, staff members can enhance their commitment to caring for their profession and their communities. PMID- 9337468 TI - Surgical glove failures in clinical practice settings. AB - Health care personnel often pay little attention to the barrier effectiveness of the surgical gloves they use in clinical settings. They may assume that all surgical gloves provide adequate protection against the transfer of bloodborne pathogens, chemicals, or mutagenic substances. Perioperative staff members frequently are unaware that their surgical gloves have failed until they find blood on their hands after operative procedures are completed. In this first article of a three-part series, the authors review current surgical glove testing standards, define surgical glove failure, and describe the reasons that surgical glove failure occurs in clinical practice settings. PMID- 9337469 TI - Introducing a music program in the perioperative area. AB - Music can touch patients deeply and thus transform their anxiety and stress into relaxation and healing. Patients with cancer who undergo surgical procedures are highly stressed. To help alleviate these patients' stress and improve their comfort, perioperative nurses at Memorial Sloan-Kettering Cancer Center (MSKCC), New York, surveyed surgical patients and staff members about introducing a perioperative music program. This article reviews the literature on the use of music in perioperative care settings and describes MSKCC's decision to evaluate and then implement a music program. PMID- 9337470 TI - Radio-frequency lesioning to treat chronic lumbar facet joint pain. PMID- 9337471 TI - Identifying risk factors for and preventing hip fractures in elderly patients. AB - Changing the mind-set of Americans to exercise more, eat right, and be responsible for their own health throughout the continuum of life is a challenge for all health care professionals. Through personal interaction with patients and participation in hospitalwide or communitywide education programs, perioperative nurses play a key role in educating the public about prevention of hip fractures in elderly patients. PMID- 9337472 TI - Designing an OR procedure book for small health care facilities. PMID- 9337473 TI - The many roles of the perioperative nurse. PMID- 9337474 TI - The challenges and rewards of providing perioperative nursing care for patients with physical and mental disabilities. PMID- 9337475 TI - Advance directives in the perioperative setting. PMID- 9337476 TI - Telehealth a complex issue being addressed by state and federal governments. PMID- 9337477 TI - Selecting samples for research studies requires knowledge of the populations of interest. PMID- 9337478 TI - Guidelines for the management of latex allergies and safe use of latex in perioperative practice settings. PMID- 9337479 TI - Oxidative damage and fragmentation of mitochondrial DNA in cellular apoptosis. AB - The molecular genetics and bioenergetics of oxidative damage, fragmentation, and fragility of mitochondrial DNA in cellular apoptosis is reviewed in connection with the "redox mechanism of ageing." PMID- 9337480 TI - The non-Ohmic proton leak--25 years on. AB - The proton conductance of the mitochondrial inner membrane can be quantified by applying Ohm's law to the experimentally determined protonmotive force and the proton current flowing around the proton circuit in the absence of ATP synthesis or ion transport. This last parameter is derived from the rate of State 4 respiration multiplied by the H+/O stoichiometry for the substrate. When the activity of the dehydrogenase supplying electrons to the respiratory chain is progressively increased the proton conductance increases rapidly when the protonmotive force is greater than 200 mV. The consequences of this non-ohmic relationship are discussed. PMID- 9337481 TI - Generating, partitioning, targeting and functioning of superoxide in mitochondria. AB - Recently, we proposed a hypothetical model of coexistence of "Reactive oxygen cycle" with Q cycle and H+ cycle in mitochondrial respiratory chain to combine both processes of univalent electron leak for production of superoxide and of proton leak across inner mitochondrial membrane. This review presents a more detailed description of this model and summaries the supporting experimental evidence obtained. PMID- 9337482 TI - "Mild" uncoupling of mitochondria. AB - Recently, it was proposed that the thyroid hormone-mediated uncoupling in mitochondria is involved in the cellular defence system against free radicals (Skulachev V.P. (1996) Quart. Rev. Biophys. 29:169-202). This phenomenon was named "mild" uncoupling. It was postulated to be a protein-mediated process controlled by several factors. The data reported during the past 40 years, pointing to the protein-mediated uncoupling mechanism in mitochondria, are reviewed in a context of hypothetical properties of "mild" uncoupling. The mechanism of "mild" uncoupling is suggested to be the following: (a) mitochondria possess protein(s) that regulate the proton permeability of inner mitochondrial membrane; (b) these proteins are regulated by binding of an unidentified low molecular-weight endogenous compound with properties resembling those of the most active artificial uncouplers like FCCP and SF6847; (c) the interaction of this compound with its target protein(s) is modulated by a thyroid hormone in a positive (i.e. enhancing the proton permeability) way and by sex steroid hormones in a negative way; (e) endogenous fatty acids can attenuate the influence of both thyroid and steroid hormones. PMID- 9337483 TI - Mitochondrial oxygen radical formation during reductive and oxidative stress to intact hepatocytes. AB - After simple respiratory inhibition, glycolytic substrates prevent cell death by providing an alternate source of cellular ATP. When mitochondrial uncoupling ensues, the uncoupler-stimulated mitochondrial ATPase hydrolyzes ATP formed by glycolysis and protection is lost. Electron transfer components abnormally reduced by respiratory inhibition, especially ubisemiquinone, react directly with oxygen to form toxic radicals. Mitochondria also generate reactive oxygen species after exposure to oxidant chemicals. A consequence is onset of the mitochondrial permeability transition, which leads to uncoupling, cellular ATP depletion and loss of viability. Thus, mitochondria are both a source and a target of toxic oxygen radicals in cell injury. PMID- 9337484 TI - Redox regulation of the mitochondrial permeability transition pore. AB - The recent data on redox regulation of the mitochondrial cyclosporin-sensitive pore are reviewed here. They indicate that the pore is modulated by the redox state of pyridine nucleotides and glutathione at two independent sites. Special attention is paid to experimental approaches for studying this phenomenon in isolated mitochondria. The relation between oxidative stress and the opening of the mitochondrial pore in some cases of cell injury and in programmed cell death (apoptosis) is discussed. PMID- 9337485 TI - Respiratory protection of nitrogenase activity in Azotobacter vinelandii--roles of the terminal oxidases. AB - Nitrogen fixation by aerobic prokaryotes appears paradoxical: the nitrogen-fixing enzymes--nitrogenases--are notoriously oxygen-labile, yet many bacteria fix nitrogen aerobically. This review summarises the evidence that cytochrome bd, a terminal oxidase unrelated to the mitochondrial and many other bacterial oxidases, plays a crucial role in aerotolerant nitrogen fixation in Azotobacter vinelandii and other bacteria by rapidly consuming oxygen during uncoupled respiration. We review the pertinent properties of this oxidase, particularly its complement of redox centres, the catalytic cycle of oxygen reduction, the affinity of the oxidase for oxygen, and the regulation of cytochrome bd gene expression. The roles of other oxidases and other mechanisms for limiting damage to nitrogenase are assessed. PMID- 9337486 TI - Structure and function of the plant alternative oxidase: its putative role in the oxygen defence mechanism. AB - Current understanding of the structure and function of the plant alternative oxidase is reviewed. In particular, the role of the oxidase in the protection of tissues against oxidative stress is developed. PMID- 9337487 TI - How do bacteria avoid high oxygen concentrations? AB - Bacteria, such as Escherichia coli and Azospirillum brasilense, avoid microenvironments with elevated oxygen concentrations, not by sensing reactive oxygen derivatives, but by sensing a metabolic down-shift that results from elevated oxygen levels. A novel protein, Aer, and the chemotaxis serine receptor, Tsr, have recently been identified as transducers for aerotaxis which monitor internal energy levels in the bacteria. PMID- 9337488 TI - The terminal oxidase in the marine bacterium Pseudomonas nautica 617. AB - The molecular properties of a novel membrane quinol oxidase from the marine bacterium Pseudomonas nautica 617 are presented. The protein contains 2b hemes/mole which may be distinguished by EPR spectroscopy but not by optical spectroscopy and electrochemistry. Respiration, though being cyanide insensitive, is not inhibited by carbon monoxide and oxygen reduction is carried out only half way with production of hydrogen peroxide. The terminal oxidase represents, therefore, a unique example in the large family of terminal oxidases known up to date. PMID- 9337489 TI - Membrane-linked systems preventing superoxide formation. AB - New facts and ideas concerning the membrane-linked mechanisms preventing superoxide formation are summarised here. It is assumed that aerobic cells possess several lines of anti-ROS defence, including optimisation of the intracellular oxygen concentration, decrease in the concentration and life-time of one-electron O2 reductants such as CoQH; and mitochondrial and cell selections, i.e. elimination of mitochondria and cells producing ROS at high rate. It is postulated that ROS-dependent pore opening and ROS-dependent apoptosis are involved in mitochondrial and cell selections. PMID- 9337491 TI - Cognitive and behavioral treatment of pathological gambling: a controlled study. AB - This study evaluated the efficacy of a cognitive-behavioral treatment package for pathological gambling. Twenty-nine men who met criteria for pathological gambling in accordance with the Diagnostic and Statistical Manual of Mental Disorders (3rd ed., revised; DSM-III-R; American Psychiatric Association, 1987) were randomly assigned to treatment or wait-list control. The treatment included 4 components: (a) cognitive correction of erroneous perceptions about gambling, (b) problem solving training, (c) social skills training, and (d) relapse prevention. The dependent variables were the South Oaks Gambling Screen, perception of control, frequency of gambling, perceived self-efficacy, desire to gamble, and number of DSM-III-R criteria met by participants. Posttest results indicated highly significant changes in the treatment group on all outcome measures, and analysis of data from 6- and 12-month follow-ups revealed maintenance of therapeutic gains. Recommendations for clinical interventions are discussed and focus on the cognitive correction of erroneous perceptions of gambling. PMID- 9337492 TI - Perceived benefit and mental health after three types of disaster. AB - The study of growth and perceived benefit after traumatic events has been hailed as one of the most promising directions for stress research. This research, however, has been limited by several methodological limitations. These limitations are addressed in this prospective study, which examines perceived benefit and mental health adjustment after 3 different types of disaster. Survivors of a tornado in Madison, Florida, had the highest rates of perceived benefit, followed by survivors of a mass killing in Killeen, Texas, and survivors of a plane crash in Indianapolis, Indiana. Perceived benefit 4-6 weeks postdisaster predicted posttraumatic stress disorder 3 years later. Perceived benefit moderated the effect of severity of disaster exposure on mental health diagnosis change over time. Without perceived benefit, as exposure severity increased, the amount of recovery decreased. If benefit was perceived, as exposure severity increased, the amount of recovery increased. Implications for clinical interventions and future research are discussed. PMID- 9337490 TI - Cognitive-behavioral treatment for depression in alcoholism. AB - Alcoholics with depressive symptoms score > or = 10 on the Beck Depression Inventory (A.T. Beck, C. H. Ward, M. Mendelson, J. Mock, & J. Erbaugh, 1961) received 8 individual sessions of cognitive-behavioral treatment for depression (CBT-D, n = 19) or a relaxation training control (RTC; n = 16) plus standard alcohol treatment. CBT-D patients had greater reductions in somatic depressive symptoms and depressed and anxious mood than RTC patients during treatment. Patients receiving CBT-D had a greater percentage of days abstinent but not greater overall abstinence or fewer drinks per day during the first 3-month follow-up. However, between the 3- and 6-month follow-ups, CBT-D patients had significantly better alcohol use outcomes on total abstinence (47% vs. 13%), percent days abstinent (90.5% vs. 68.3%), and drinks per day (0.46 vs. 5.71). Theoretical and clinical implications of using CBT-D in alcohol treatment are discussed. PMID- 9337493 TI - A comparative analysis of the therapeutic focus in cognitive-behavioral and psychodynamic-interpersonal sessions. AB - This study compared therapeutic foci in a sampling of 30 cognitive-behavioral and 27 psychodynamic-interpersonal manual-driven treatments for depression. High- and low-impact sessions were coded for each client, with the Coding System of Therapeutic Focus. Results indicated that psychodynamic-interpersonal sessions focused more on such variables as emotion, patterns, incongruities, the impact that others made on clients, clients' expected reaction of others, the tendency to avoid therapeutic progress, therapists themselves, clients' parents, and links between people and time periods in clients' lives. Cognitive-behavioral sessions placed greater emphasis on external circumstances and clients' ability to make decisions, gave more support and information and encouraged between-session experiences, and focused more on the future. Relatively few differences emerged as a function of session impact. Results are discussed in terms of the different and similar theoretical conceptions of the change process. PMID- 9337494 TI - Social skills training with parent generalization: treatment effects for children with attention deficit disorder. AB - The effectiveness of brief social skills training (SST) was evaluated in a controlled outcome study with 27 children meeting criteria of the Diagnostic and Statistical Manual of Mental Disorders (3rd ed., revised; American Psychiatric Association, 1987) for an attention deficit disorder. Children were randomly assigned to either SST with parent-mediated generalization (SST-PG), child-only SST, or a wait-list control group. SST consisted of 8 group sessions in which skill modules were taught sequentially. Parents of children in the SST-PG group simultaneously participated in group generalization training designed to support their children's transfer of skills. Significant improvement in children's skill knowledge and in parent reports of social skills and disruptive behavior occurred for both treatment groups relative to the wait-list control group and maintained at a 4-month follow-up. More modest evidence was found for generalization of SST to the school setting. PMID- 9337495 TI - Adolescent outcome of boys with attention-deficit/hyperactivity disorder and social disability: results from a 4-year longitudinal follow-up study. AB - IQ-achievement discrepancy methodology similar to that used in defining learning disabilities has recently been used to identify a subset of boys with attention deficit/hyperactivity disorder (ADHD) evidencing marked impairment in social functioning. In this study, 2 issues were examined: (a) What is the longitudinal outcome of boys with ADHD identified at baseline as "socially disabled"? (b) Is social disability at baseline a significant predictor of severe long-term outcomes (such as substance use disorders) in boys with ADHD? If so, are its predictive relationships accounted for by conditions that are comorbid with ADHD? Results showed that, at follow-up, boys with ADHD who also had social disability evidenced significantly higher rates of mood, anxiety, disruptive, and substance use disorders, compared with nonsocially disabled boys with ADHD and comparison boys without ADHD. Findings also showed that social disability at baseline in boys with ADHD was a significant predictor of later conduct disorder and most substance use disorders after baseline mood and conduct disorders and behavior checklist ratings of aggressive behavior and attention problems were controlled. PMID- 9337496 TI - Affiliation with Alcoholics Anonymous after treatment: a study of its therapeutic effects and mechanisms of action. AB - Relatively little is known about how substance abuse treatment facilitates positive outcomes. This study examined the therapeutic effects and mechanisms of action of affiliation with Alcoholics Anonymous (AA) after treatment. Patients (N = 100) in intensive 12-step substance abuse treatment were assessed during treatment and at 1- and 6-month follow-ups. Results indicated that increased affiliation with AA predicted better outcomes. The effects of AA affiliation were mediated by a set of common change factors. Affiliation with AA after treatment was related to maintenance of self-efficacy and motivation, as well as to increased active coping efforts. These processes, in turn, were significant predictors of outcome. Findings help to illustrate the value of embedding a test of explanatory models in an evaluation study. PMID- 9337497 TI - Group counseling versus individualized relapse prevention aftercare following intensive outpatient treatment for cocaine dependence: initial results. AB - Ninety-eight male cocaine-dependent patients who completed an intensive outpatient program (IOP) were randomly assigned to either standard group counseling (STND) or individualized relapse prevention (RP) aftercare. Heavier cocaine and alcohol use during IOP and low self-efficacy predicted more cocaine use during the treatment phase of the study, whereas lifetime diagnoses of alcohol dependence, major depression, and any anxiety disorder predicted less cocaine use. Rates of complete abstinence during the 6-month study period were higher in STND than RP, whereas RP was more effective in limiting the extent of cocaine use in those who used during Months 1-3. Matching analyses indicated patients who failed to achieve remission from cocaine dependence during IOP and those with a commitment to absolute abstinence did better in RP than in STND, whereas patients with other abstinence goals did better in STND than RP. Several differences in experiences before cocaine use and "near-miss" episodes were also identified. PMID- 9337498 TI - Behavioral couples therapy for male substance-abusing patients: a cost outcomes analysis. AB - The cost outcomes for married or cohabiting substance-abusing male patients (N = 80) who were randomly assigned to receive either behavioral couples therapy (BCT) or individual-based treatment (IBT) were compared. Social costs incurred by patients in several areas (e.g., cost of substance abuse treatment, support from public assistance) during the year before and the year after treatment were estimated. BCT was more cost-beneficial than IBT; although the monetary outlays for delivering IBT and BCT were not different, the average reduction in aggregate social costs from baseline to follow-up was greater for patients who received BCT (i.e., $6,628) than for patients who received IBT (i.e., $1,904). BCT was also more cost-effective than IBT; for each $100 spent on the treatment, BCT produced greater improvements than IBT on several indicators of treatment outcome (e.g., fewer days of substance use, fewer legal problems). PMID- 9337500 TI - Traumatic events: prevalence and delayed recall in the general population. AB - A random sample of 724 individuals across the United States were mailed a questionnaire containing demographic information, an abridged version of the Traumatic Events Survey (D. M. Elliott, 1992), and questions regarding memory for traumatic events. Of these, 505 (70%) completed the survey. Among respondents who reported some form of trauma (72%), delayed recall of the event was reported by 32%. This phenomenon was most common among individuals who observed the murder or suicide of a family member, sexual abuse survivors, and combat veterans. The severity of the trauma was predictive of memory status, but demographic variables were not. The most commonly reported trigger to recall of the trauma was some form of media presentation (i.e., television show, movie), whereas psychotherapy was the least commonly reported trigger. PMID- 9337499 TI - Effects of adding behavioral treatment to opioid detoxification with buprenorphine. AB - This trial assessed whether behavioral treatment improves outcome during a 26 week outpatient opioid detoxification. Thirty-nine opioid-dependent adults were assigned randomly to a buprenorphine dose-taper combined with either behavioral or standard treatment. Behavioral treatment included (a) a voucher incentive program for providing opioid-free urine samples and engaging in verifiable therapeutic activities and (b) the community reinforcement approach, a multicomponent behavioral treatment. Standard treatment included lifestyle counseling. Fifty-three percent of the patients receiving behavioral treatment completed treatment, versus 20% receiving standard treatment. The percentage of patients achieving 4, 8, 12, and 16 weeks of continuous opioid abstinence were 68, 47, 26, and 11 for the behavioral group and 55, 15, 5, and 0 for the standard group, respectively. Behavioral treatment improved outcomes during outpatient detoxification. PMID- 9337501 TI - Multisystemic therapy with violent and chronic juvenile offenders and their families: the role of treatment fidelity in successful dissemination. AB - The effects of multisystemic therapy (MST) in treating violent and chronic juvenile offenders and their families in the absence of ongoing treatment fidelity checks were examined. Across 2 public sector mental health sites, 155 youths and their families were randomly assigned to MST versus usual juvenile justice services. Although MST improved adolescent symptomology at posttreatment and decreased incarceration by 47% at a 1.7-year follow-up, findings for decreased criminal activity were not as favorable as observed on other recent trials of MST. Analyses of parent, adolescent, and therapist reports of MST treatment adherence, however, indicated that outcomes were substantially better in cases where treatment adherence ratings were high. These results highlight the importance of maintaining treatment fidelity when disseminating complex family based services to community settings. PMID- 9337502 TI - A 2-year longitudinal analysis of the relationships between violent assault and substance use in women. AB - Women experience alarming levels of physical and sexual assault, which may lead to escalation of substance use. Reciprocally, evidence from cross-sectional studies indicates that substance use may increase risk of assault. To date, directionality of this relationship remains unclear. This issue is addressed by the present 3-wave longitudinal study in which a national probability sample of 3,006 women were followed for 2 years. Dependent measures were obtained at each wave of the study and included questions about lifetime and new assault status, alcohol abuse, and drug use. Wave 1 use of drugs, but not abuse of alcohol, increased odds of new assault in the subsequent 2 years. Reciprocally, after a new assault, odds of both alcohol abuse and drug use were significantly increased, even among women with no previous use or assault history. For illicit drug use, findings support a vicious cycle relationship in which substance use increases risk of future assault and assault increases risk of subsequent substance use. PMID- 9337503 TI - Psychopathy and sexual assault: static risk factors, emotional precursors, and rapist subtypes. AB - This study compared psychopathic and nonpsychopathic rapists on static risk factors and on emotional and motivational precursors. Sixty incarcerated rapists were assessed for psychopathy with the Psychopathy Checklist--Revised (R. D. Hare, 1991), and they were classified according to the Massachusetts Treatment Center: Revised Rapist Typology, Version 3 (R. A. Knight & R. A. Prentky, 1990b). Psychopathy was positively associated with past nonsexual offenses and negatively associated with age onset for criminal offending, number of sexual victims, and the intensity of negative emotions experienced before sexual offending. However, psychopathy was not related to sexual offense history, age of onset for sexual offending, or victim harm. Last, psychopaths were most likely to be classified as opportunistic and pervasively angry rapists. The findings indicate that psychopathy should be considered when developing intervention strategies for rapists. PMID- 9337504 TI - MMPI-2 profiles of adult women with child sexual abuse histories: cluster analytic findings. AB - This study used a cluster analysis to examine the clinical profiles of female survivors of child sexual abuse. Eighty-five participants who presented for group therapy to deal specifically with issues related to sexual abuse completed the revised version of the Minnesota Multiphasic Personality Inventory (MMPI-2; J. N. Butcher, W. G. Dahlstrom, J. R. Graham, A. Tellegen, & B. Kaemmer, 1989) as part of an extensive assessment procedure. The cluster-analytic procedure used in this study allowed 5 subgroups within the population to emerge, supporting the idea that women who report having been sexually abused as children are not a homogeneous group. Additional analyses indicated differences on the basis of cluster membership on the MMPI-2 content scales, as well other measures of psychological distress. The treatment implications of these findings are discussed. PMID- 9337505 TI - Mastery and inoculation against setbacks as active ingredients in the JOBS intervention for the unemployed. AB - Earlier analyses of the JOBS II intervention for unemployed job seekers demonstrated that the intervention facilitated reemployment, reduced depressive symptoms, and improved role and emotional functioning (A. D. Vinokur, R. H. Price, & Y. Schul, 1995). The present study focuses on mediational processes of the active ingredients targeted by the intervention. Structural equation modeling analysis demonstrated that an enhanced sense of mastery had mediating effects of reemployment, financial strain, and reduction in depressive symptoms. Another active ingredient, inoculation against setbacks, was shown to protect those who suffered the setback of losing a job after temporarily regaining one. The inoculation protected them from experiencing the high level of depressive symptoms that was typical of their counterparts in the control group. PMID- 9337506 TI - Cue exposure in moderation drinking: a comparison with cognitive-behavior therapy. AB - To date, the published controlled trials on exposure to alcohol cues have had an abstinence treatment goal. A modification of cue exposure (CE) for moderation drinking, which incorporated priming doses of alcohol, could train participants to stop drinking after 2 to 3 drinks. This study examined the effects of modified CE within sessions, combined with directed homework practice. Nondependent problem drinkers who requested a moderation drinking goal were randomly allocated to modified CE or standard cognitive-behavior therapy (CBT) for alcohol abuse. Both interventions were delivered in 6 90-min group sessions. Eighty-one percent of eligible participants completed treatment and follow-up assessment. Over 6 months, CE produced significantly greater reductions than CBT in participants' reports of drinking frequency and consumption on each occasion. No pretreatment variables significantly predicted outcome. The modified CE procedure appears viable for nondependent drinkers who want to adopt a moderate drinking goal. PMID- 9337507 TI - Attrition in the treatment of childhood anxiety disorders. AB - The present study explored the differences between completers and terminators (including both refusers and dropouts) of an individual cognitive-behavioral treatment for childhood anxiety. Participants were 190 children with anxiety disorders and their parents: 146 completed treatment and 44 terminated. Terminators were more likely to live in a single-parent household, be ethnic minorities, and self-report less anxious symptomatology. Follow-up interviews indicated that identifiable child factors were influential in terminators' decisions to discontinue treatment. Among terminators, differences between refusers and dropouts were also investigated. PMID- 9337508 TI - Aptitude-treatment interactions based on clients' assimilation of their presenting problems. AB - Assigning clients to treatments on the basis of their differential aptitudes for those treatments may reduce variability and improve the mean outcomes of psychotherapy. The assimilation model suggests that in time-limited treatments, clients with well-assimilated problems would do better in cognitive or behavioral therapies than in psychodynamic, experiential, or interpersonal therapies, whereas the reverse should be the case for clients with poorly assimilated problems. Results for high-, moderate-, and low-assimilation subgroups (based on rating the level of assimilation of problems presented in the first 20 min of first sessions) of clients (N = 112) randomly assigned to time-limited cognitive behavioral or psychodynamic-interpersonal treatment supported the first suggestion but not the second (clients with poorly assimilated problems did equally well in both treatments). PMID- 9337510 TI - Analyses of factors influencing participation in the cervical cancer screening programme in the community in Japan. AB - The association between the participation rates of cervical cancer screening programme and implementation methods, concerning data management, participant convenience, promoting participation, and payment, were analyzed. The data regarding the implementation methods were obtained in a nation-wide questionnaire survey. The relationship between the participation rates and implementation methods were assessed using the chi-square test and multiple logistic regression analyses. In small municipalities, with a population < 10,000 items concerning data management, enlisting the assistance of community organizations, fee exemption, and early morning screening were positively associated with the participation. In middle-sized municipalities, with the population 10,000-49,999, early morning screening, community organizations, items concerning data management, and sending out letters were positively associated with participation. Saturday/Sunday screening, community organizations, letters, and postcards were positively associated with the participation of the older group (> or = 50 years) in large municipalities with population of > 50,000. These results indicate that enlisting the assistance of community organizations and establishing a well-organized data management system are likely to improve participation regardless of municipality size. Other implementation methods must be selected taking into account factors such as municipality population size, as well as the age distribution and characteristics of the target population. PMID- 9337509 TI - Risk factors associated with uterine cervical cancer in Korea: a case-control study with special reference to sexual behavior. AB - OBJECTIVE: A hospital-based case-control study was conducted to identify characteristics of women at high risk of developing cervical cancer with special reference to sexual behavior in Korea. METHODS: Histologically confirmed cases of invasive cervical cancer were selected from the Department of Gynecology, Seoul National University Hospital between September 1992 to May 1995 (n = 203). Women with normal pap smear tests and women free of past history of any malignancies were regarded as controls (n = 827). Information on risk factors were collected by both a self-administered questionnaire and a direct interview. RESULTS: Uterine cervical cancer risk was higher in women with a less educated spouse (Ptrend = 0.0003), women with a family history of cervical cancer (adjusted odds ratio [OR] = 2.20., 95% confidence interval [CI] 1.21-4.01), women of shorter height (Ptrend = 0.02), women with early age at first full term pregnancy (Ptrend = 0.0005), and women who have had multiple full term pregnancies (Ptrend = 0.006) by the multiple linear logistic analysis. Particularly noteworthy was a significant decreasing trend in the adjusted OR with the age at first sexual intercourse increasing (Ptrend = 0.002) after adjusting the number of sexual partners. The husband's indecent sexual history showed a borderline significance (Ptrend = 0.07). CONCLUSIONS: This study confirmed that the risk factors of cervical cancer in Korea are similar with those found in other countries. PMID- 9337511 TI - Cost-effective analysis of mass screening for cervical cancer in Japan. AB - Cost-effectiveness analysis for cervical cancer screening in Japan was performed to estimate the cost per life-year saved by the screening; cost-effectiveness ratio (CER). The analysis was made using a simulation model to estimate long-term cost and effectiveness of the screening programs. CER of cervical cancer screening was estimated to be US$ 40,604 which was 2.4 times more expensive than that for gastric cancer screening but was about the same as that for colorectal cancer screening. It was within the range of cost-effectiveness of other cancer screening programs financed under the Health and Medical Services Law for the Aged in Japan. We performed sensitivity analysis on the following seven estimates, the screening charge, the sensitivity and the specificity of the screening test, the frequency of carcinoma in situ (CIS) among cases detected in the screening program, the initial cost and the terminal cost for patients with invasive cancer, and the incidence rate of cervical cancer. The sensitivity analysis demonstrated that the screening charge was the most influential factor on CER. CER was fairly stable under various assumptions on the accuracy of the screening test, the frequency of carcinoma in situ (CIS), the treatment cost for patient, and the incidence of cervical cancer. CER was less sensitive to the changes in incidence, even to as low as a 50% decrease of the current figure. Then if the incidence rate becomes 85% of the current figure in 2015, CER would be US$ 48,176 and it was suggested that the cervical cancer screening would remain reasonably cost-effective until the year 2015. PMID- 9337512 TI - Evaluation of cutoff levels for screening of gastric cancer using serum pepsinogens and distributions of levels of serum pepsinogen I, II and of PG I/PG II ratios in a gastric cancer case-control study. AB - This study was carried out to determine the cutoff levels of serum pepsinogen (PG) I, II and their ratio of PG I/PG II for gastric cancer to establish a better screening system. Optimal cutoff levels for gastric cancer screening using serum pepsinogens were determined using Youden's index. The sensitivity, specificity and Youden's index for gastric cancer cases were calculated according to sex, age and the stage of gastric cancer, and the maximum Youden's index in each category was adopted as the cutoff level for gastric cancer screening using serum pepsinogens. The maximal Youden's index in all gastric cancer cases was 0.37, corresponding to a cutoff level of PG I < 40 (micrograms g/l) and PG I/PG II < 3.5. The sensitivity and specificity for gastric cancer cases of these cutoff levels were 0.50 and 0.87, respectively. In future, better criteria for gastric cancer screening have to be examined with the estimation of Youden's index in addition to other epidemiological methods such as ROC (receiver operating characteristic) curves and/or cost benefit analyses. PMID- 9337513 TI - Effects of misclassification and temporal change of response in food frequency on risk ratio. AB - Misclassification and temporal changes of food consumption frequencies were estimated under the statistical models with some assumptions, and their effects on risk ratios were evaluated. Food frequencies of 27 items in 214 subjects were doubly measured by a questionnaire with 2 weeks interval, and those in 326 subjects were measured in 1989, 1993 and 1994. Median of probabilities of misclassification in responses among 27 food items was estimated to be 0.12. Medians of proportions of persons whose responses in 1989 were different from those after 5 years, were calculated to be 35% with misclassification and 27% without misclassification. For the true risk ratio of 3, medians of the risk ratios of dietary habits during 5 years, based on food frequencies measured at 1989, were observed to be 2.2 in case of responses with misclassification, and 1.7 in case of responses with misclassification and temporal changes. These suggested that the risk ratios of food frequencies would be seriously affected by misclassification and temporal changes in responses. PMID- 9337514 TI - Anticipation of job loss or job change and cardiovascular risk factors: a study of retiring self-defense officials in Japan. AB - Self-defense officials in Japan are to retire at the age of early 50s. This unique situation prompted the authors to investigate whether preexisting morbid conditions cause any difficulty in finding a post-retirement job and whether anticipation of job loss or job change, as measured by the status of post retirement job and months remaining until retirement, was related to biological cardiovascular risk factors. The subjects were 2,228 male self-defense officials who received a preretirement health examination at three Self-Defense Forces Hospitals from 1991 to 1992; the period in time remaining until retirement ranged from 1-40 months (median 12 months), and 62% had one year or less until the retirement. The defined preexisting illnesses included a wide range of chronic, non-communicable diseases. Overall, the preexisting illness was unrelated to the determination of a post-retirement job. In men having 6 months or less until retirement, however, the security of post-retirement job was less frequent when they had the preexisting illness, especially cardiovascular diseases. In 1,839 men excluding those with the preexisting illness, the period until retirement was not adversely related to obesity, blood pressure, serum lipids, serum uric acid, or glucose intolerance whether the post-retirement job had been secured or not. The findings suggest that the preexisting illness decreases the chance of obtaining a post-retirement job, but do not provide any evidence that anticipation of job loss or job change due to early retirement exerts an adverse effect on biological cardiovascular risk factors. PMID- 9337515 TI - Methicillin-resistant Staphylococcus aureus (MRSA) isolation from pharyngeal swab cultures of Japanese elderly at admission to a geriatric hospital. AB - The isolation rate of the methicillin-resistant Staphylococcus aureus(MRSA) from pharyngeal swab cultures in Japanese elderly was studied at admission to a geriatric hospital. The subjects were 233 consecutive patients admitted to Kitakyushu Tsuyazaki Hospital from April 1994 to March 1996. The isolation rate of MRSA was 3.0% in the patients admitted from their own homes, 9.7% in those transferred from nursing homes and 14.0% in those transferred from other hospitals. The patients from their own homes were younger than those from nursing homes, the latter being older than those transferring from other hospitals. The patients from their own homes had better activities of daily living(ADL), higher levels of hemoglobin and serum albumin than those from nursing homes or other hospitals. The white blood cell counts, and the proportion of patients with positive c-reactive protein or with fever did not differ among the three groups. Multiple logistic regression analysis revealed that fever and ADL disability were independent risk factors for the isolation of MRSA, and hypoalbuminemia was a risk factor for MRSA isolation in the model using serum albumin instead of ADL score. These results suggest that the lower isolation rate MRSA among patients from their own homes may be partly due to better ADL and nutritional status compared with those from nursing homes or other hospitals. PMID- 9337516 TI - Follow-up study of children with precocious puberty treated with cyproterone acetate. Ad hoc Committee for CPA. AB - A total of 1840 children and adolescents treated with cyproterone acetate (CPA) to block gonadal function, as a treatment for precocious puberty, short stature and other disorders, were registered to survey for the risk of developing hepatic tumors. Patients responding to follow-up numbered 1552 (85%). The cumulative dose and duration of CPA therapy for boys and girls were 110.4g and 2.6 years, and 122.9 g and 2.8 years, respectively. Among the 1552 patients, five hepatoma cases were found. Four underwent successful surgery and remain alive and well to date. Two of the 5 cases had been given more than 500g, the other 3 more than 1000 g, of CPA. Three had also been given androgens before CPA administration. Although further follow-up is necessary to monitor for the development of adenoma and hepatoma, the risk of developing these tumors among patients to whom limited doses of CPA were administered appears to be negligible. PMID- 9337517 TI - Hepatitis C virus and human T cell leukemia virus type 1 infection without [corrected] blood transfusion in hemodialysis units and its prevention. PMID- 9337518 TI - Oncogenes and tumour-suppressor genes in lung cancer. PMID- 9337519 TI - Cytokines in lung cancer. AB - Cytokines are hormonE-like proteins and peptides involved in the signalling between cells during immune response. They are produced mainly by lymphocytes (lymphokines) and mononuclear phagocytes (monokines). They are involved in both cell-mediated and humoral immunity. Cytokines fall into a number of categories: interferons (IFNs), interleukins (ILs) and growth factors. It has been indicated in cancer immunology the following cytokines are particularly important: IFNs, TNF-alpha, IL-1, IL-2, IL-4, IL-6, IL-7, IL-9, IL-10, IL-12. Interferons (IFN gamma in particular) in cooperation with TNF-alpha and IL-1 inhibit proliferation of tumor cells and by their synergic activity with IL-2 induce cytotoxicity of NK cells. They activate mononuclear phagocytes and by B lymphocyte stimulation augment lysis of cancer cells. TNF-alpha has mainly cytotoxic activity, leading to hemorrhagic necrosis of tumors. It is also an endogenic pyrogen which is together with IL-1 responsible for pyretic status in neoplastic disease. IL-1 stimulates necrotizing activity of TNF-alpha and augments cachexia by anabolism of lipid induction. IL-2, IL-6 and IL-12 induce NK and LAK-cell cytotoxicity. IL 12 inhibits metastasis formation. IL-10, by inhibiting synthesis of cytokines may lead to tumor development. PMID- 9337520 TI - The value of immunohistochemical identification of neuroendocrine differentiation in non small cell lung carcinoma. AB - Neuroendocrine differentiation (NE) has been found to be a feature of neuroendocrine tumours and also a proportion of non-small cell lung carcinomas. Detection of NE by immunohistochemical assays depends on spectrum of used antibodies and criteria. Immunohistochemical identification of NE in non-small cell lung cancer has been discussed in an article together with criteria, diagnostic helpfulness and prognostic value of this feature. PMID- 9337521 TI - Pulmonary carcinoid and related tumours. AB - Since the turn of the century carcinoid tumours have fascinated clinicians and pathologists. It may be because of their unpredictable behaviour, characteristic morphology or the unusual nature of the carcinoid syndrome. Whatever the reason, in the last twenty years the topic of neuroendocrine features in a tumour has been to the fore in pathology, probably because small cell carcinoma is so sensitive to chemo- and radiotherapy. This article will review the clinical and pathological features of typical and atypical carcinoid tumours. The relative "newcomer" of large cell neuroendocrine carcinoma will also be described. No description of small cell carcinoma will be given as it is available in standard texts and recent reviews [1, 2]. A classification of the common neuroendocrine tumours is given in Tab. I. PMID- 9337522 TI - Extralysosomal degradation of proteins. AB - Extralysosomal degradation of proteins is carried out by proteases complex (proteasomes), calcium dependent calpains, proteases of the rough endoplasmatic reticulum and proteases of the cellular membrane. It depends on a limited proteolysis and includes the main enzymatic proteins, hormones, growth factors and cytoskeletal proteins. Thus it plays an important regulatory role in the metabolism and formation of cellular structures. PMID- 9337523 TI - Proteolytic enzymes in proliferation and neoplastic metastases formation. AB - Metalloproteases, plasminogen urokinase activator, plasmin and cathepsins enable the expansion of neoplastic tumors, leading to metastases formation. They cause neoplastic cells to detach from tumor, facilitate cell movement, implantation and participate in tumor vascularization. The regulation of these processes is accomplished during the synthesis and activation of proenzymes. Enzyme activity control is realized by their bonds with cellular membranes, and inhibitor action. PMID- 9337524 TI - Lysosomal cysteine proteinases and their significance in pathology. AB - The lysosomal cysteine proteinases are synthesized in a form of pre-proenzymes. They are submitted to posttranslational glycosylation and phosphorylation. These modifications make possible transport the modified proteins into Golgi apparatus and into lysosomes. Some disturbances of transport which occur mainly in tumour cells result in an increase of these enzymes activities in cytosol and in intercellular compartment. The activity of cysteine proteinases is regulated by specific inhibitors (cystatin, kininogen) which exist in some tissues and body fluids. The evaluation of activity and concentration of proteinases and inhibitors is important in clinical diagnostics. PMID- 9337525 TI - Plasminogen activators and plasmin in lung cancer. AB - Urokinase plasminogen activator and plasmin contribute to detach neoplastic cells from solid tumor and facilitate the movement of these cells through interstitium and capillary walls as well as infiltration of the surrounding structures. Plasminogen activators inhibitors fulfill a regulatory function in these processes. Determining activity and concentration, finding subcellular, cellular and zonal localization of every component of plasminogen activation system has diagnostic and prognostic importance in different lung cancer types. PMID- 9337526 TI - Biological and diagnostic role of cathepsin D. AB - Biosynthesis, posttranslating modifications, intracellular transport and activation of procathepsin D are discussed. Active cathepsin D evokes degradation of cellular and extracellular proteins and it also activates proenzymes, prohormones and growth factors and inactivates their active forms. Impairment of lysosomes in hypoxia or necrosis evokes transition of cathepsin D to cytosol and body fluids. Increase of cathepsin D content in cytosol also evokes enhancement of the synthesis rate observed among others, in neoplastic tissues and regenerating organs. Increase of cathepsin D content and activity in cytosol and blood serum is of essential diagnostic and prognostic importance in some pathologic conditions. PMID- 9337527 TI - Angiogenesis in cancer. AB - The concept of angiogenesis and consecutive stages of the neovascularization processes under physiological and pathological conditions have been described. Angiogenesis is regulated by the different mechanisms which are in dynamic balance. The regulating components of these processes are the stimulating and inhibiting factors, the mediators of these reactions under influence of the host cell-tumor cell interaction. The role of angiogenesis in cancer development is connected with obtaining the angiogenic phenotype by tumor when the transformation from prevascular to vascular phase of neoplasm goes on. The further tumor growth and metastasis processes depend on neovascularization. Actual research trends in the field of angiogenesis have been presented in this paper. We need to know such markers of angiogenesis would be the most useful for doing research work and monitoring neoplasm diseases in clinics. Antiangiogenic management seems to be a new promising therapeutic concept in oncology. PMID- 9337528 TI - Role of endothelins in lung pathology. AB - The endothelins are a group of peptides present in a number of tissues and organs. They are powerfully vasoactive, causing both contraction and relaxation of blood vessels. They are also active in the lung causing long lasting bronchoconstriction. Endothelins binding sites are widely distributed and are localized to airway and pulmonary vascular smooth muscle, fibroblasts, submucosal glands, and airway nerves, indicating that endothelins may have widespread effects. The current study presents a potential contribution of endothelins to pathogenesis of a number of lung diseases. PMID- 9337529 TI - Cancer procoagulant--CP. AB - A review of literature concerning cancer procoagulant (CP) has been carried out. This procoagulant directly activates coagulation factor X to factor Xa. Possibilities of utilising determinations of this activator in diagnostics and prognostics of the cancerous disease are discussed. PMID- 9337530 TI - Tissue factor pathway inhibitor (TFPI). AB - Tissue factor pathway inhibitor (TFPI) is a plasma proteinase inhibitor. It is a 42 kD glycoprotein that consists of 276 amino acid residues which sequence is known. TFPI is synthesized by vascular endothelial cells and part of it is associated with glycosaminoglycans of these cells. In blood TFPI is found in a free-form (active) and in an associated with lipoprotein form (nonactive). TFPI directly inhibits activated factor Xa and then factor VIIa/TF complex. Decreasing TFPI activity facilitates an activation of blood coagulation and fibrin forming, increasing TFPI activity inhibits these processes. PMID- 9337531 TI - TNF-alpha, IL-1 and IL-6 concentration in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC). AB - Fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) provide facilities for biologically active substances directly produced by the tumor. In the present study we have investigated the concentration of the following cytokines: TNF alpha, IL-2 and IL-6 in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC) before treatment. The study group consisted of 20 patients with squamous cell carcinoma (Gr. I). The control group consisted of 18 patients with non-malignant lung disease (6 patients with sarcoidosis and 12 with COPD). All patients underwent bronchoscopy followed by bronchoalveolar lavage (BAL). The concentrations of the above mentioned cytokines were measured using Sorin's ETI system for EIA analysis. Statistical analysis was performed within the groups, between the groups, and in different stages of malignant disease. The mean TNF alpha concentration in Gr. I was 1192 pg/ml/mg p. and was significantly higher than in sarcoidosis (5.3 pg/ml/ mg p.) and COPD (0.5 pg/ml/mg p.). We observed a correlation between TNF-alpha concentration and the stage of malignant disease. The highest concentration was in IIIb stage (2150 pg/ml/mg p.). IL-6 concentration in malignant patients was strongly correlated with TNF-alpha concentration and was significantly higher than in control (265.868 pg/ml/mg p. in cancer patients, in sarcoidosis: 21.694 pg/ml/mg p. and in COPD: 40.708 pg/ml/mg p.). It was the highest in stage II (334 pg/ml/mg p.). IL-1 concentrations were not significantly higher in malignant patients (50.173 pg/ml/mg p., nor in IIIa stage (70.136) pg/ml/mg p. as compared with controls (18.648 pg/ml/mg p. in sarcoidosis and 28.395 pg/ml/mg p. in COPD). PMID- 9337532 TI - IL-2 concentration in bronchoalveolar lavage fluid (BALF) of non-small cell lung cancer (NSCLC) patients. AB - The results of treatment and the survival time of lung cancer patients are strictly dependent on early diagnosis. Fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) are the most effective diagnostic methods in cancer diagnosis. These methods allowed us to evaluate biological neoplastic markers at the site of the tumor. Using commercially available ELISA kits (Endogen) followed by ETI-system (SORIN) analysis we measured the IL-2 concentration in BALF of 36 non-small cell lung cancer (NSCLC) patients before (Gr. I) and after (Gr. II) surgery and 18 non-neoplastic lung disorder patients as a control group (6 cases of sarcoidosis and 12 cases of COPD). In BALF of Gr. I Macrophage percentage was higher (x = 74.125%), and lymphocyte percentage was lower (x = 15.875%) than in sarcoidosis x = 38.33% and x = 64.3% respectively. IL-2 of BALF was not detected in 83.4% of squamous cell lung cancer cases (Gr. I) before treatment. The average IL-2 BALF concentration of the remaining portion of this group was 80.49pg/ml/mg p. IL-2 was detected in Gr. II (x = 151.003 pg/ml/mg p.) after combined cancer resection. An inverse correlation was found between IL-2 BALF concentration and disease stage. PMID- 9337533 TI - Bronchopulmonary carcinoid tumors--a significant diagnostic problem. AB - The problems with classification and diagnosis of bronchopulmonary carcinoid and other neuroendocrine tumors are described in this paper. Single neuroendocrine cells and so-called neuroepithelial bodies found in normal bronchial epithelium are currently believed to constitute pulmonary components of an extensive neuroendocrine system. In view these opinions, it has appeared a need for a new, standardized nomenclature. Hence presently suggested classification of neuroendocrine carcinomas taking into account their histological structure, immunohistochemical as well as prognostic features starting from carcinoid tumors to small cell anaplastic carcinomas-includes three types of neoplasms. PMID- 9337534 TI - Immunohistochemical analysis of tissue localization of cytokeratin 19 in lung cancer. AB - Cytokeratins (CK) are one of the main families of intermediate filaments which make up the cytoskeleton. CK19 is strongly expressed by normal simple bronchial and respiratory epithelium as well as by their malignant counterpart. Although CK19 is a part of the cytoskeleton, a soluble fragment of this polypeptide can be released and assayed in the blood as CYFRA 21-1, new sensitive and valuable marker of non small cell lung cancer. In some cases, however, discrepancies between the serum level of CYFRA 21-1 and presence of tumor and its histological type have been observed. We studied immunohistochemically tissue localization of CK19 in tumors and non invaded lung parenchyma in a series of 34 patients surgically treated due to lung cancer. CK 19 was detected in cancer cells as well as in non neoplastic epithelium covering bronchial tree and alveolar surfaces. We found a different expression of CK19 in different histological type of tumors. The most intensive expression revealed squamous cell carcinomas and adenocarcinomas. Small cell cancer revealed poor expression of CK19. In non invaded parts of the resected lungs we found the strong expression of CK19 in the cytoplasm of regenerative II type pneumocytes occurring in large quantity in the cases of interstitial lung fibrosis concomitant with some tumors. We suggest it may be a cause of unexpectedly elevated serum levels of CYFRA 19-21 in some not oncological patients or patients with small cell lung cancer. PMID- 9337535 TI - Neuron-specific enolase (NSE) serum level in non small cell lung cancer--can it be an indicator of tumour chemosensitivity? AB - It is believed that the tissue or serum expression of neuroendocrine markers in non small cell lung cancer patients can implicate better prognosis in cases undergoing chemotherapy. The aim of the study was to assess the value of NSE serum level for anticipation of the tumor response to chemotherapy. We found elevated serum level of NSE in 34 of 60 patients (56.7%) at the time of diagnosis of inoperable non small cell lung cancer, significantly more often in those presenting stage IV of disease. In 21.7% partial response and in another 21.7% minimal regression was found after chemotherapy. Treatment results revealed no significant differences in respect to NSE serum level, however 77% of patients achieving partial response had elevated serum level of NSE. PMID- 9337536 TI - CEA, NSE and SCC Ag in bronchial lavage in patients with lung cancer. AB - CEA, NSE and SCC Ag levels were measured in bronchial lavage (BL) and serum in patients with endobronchial lung cancer (LC) and in patients with nonmalignant lung diseases (NMLD). In both groups of patients 100 ml of normal saline solution was used during the lavage procedure. Tumor markers were detected using radioimmunoassay. CEA levels in BL and in serum were measured in 84 patients with LC and in 94 patients with NMLD. BL CEA levels were significantly increased in patients with LC (97.4 +/- 56.4 ng/ml) in comparison to patients with NMLD (4.2 +/- 6.3 ng/ml). Patients with LC had CEA levels in lavage fluid about 30 times higher than in serum. Significantly increased BL CEA levels in patients with LC were found in the cases with more advanced bronchoscopic tumor and in those with positive bronchial secretion cytology than in other groups of patients with LC. NSE levels were measured in BL and in serum in 21 patients with small cell lung cancer, SCC Ag levels were measured in BL and in serum in 21 patients with squamous cell carcinoma. The control group consisted of 28 patients with NMLD and 8 patients with adenocarcinoma. Determination of CEA levels in BL can be useful additional diagnostic method in patients with LC. The measurement of NSE and SCC Ag levels in BL in LC patients was considered to be not useful because of the low diagnostic sensitivity and specificity. PMID- 9337537 TI - Trophoblastic markers and CEA in non-small cell lung cancer: the comparison studies of tumour cells expression and serum concentration. AB - The study was designed to explore the tumour cell expression of three markers: hCG, SP1 and CEA in lung cancer in relationship with histological type and histological differentiation of tumours as well as serum concentration of antigens. 58 primary resected lung cancers: 28 adenocarcinomas, 27 squamous cell and 3 large cell carcinomas were included. Tumours immunoreactivity of three markers was evaluated by semiquantitative analysis. Simultaneously serum tumour markers were measured in 42 patients by enzyme radioimmunoassays. CEA and SP1 expressions in lung tumours were found in a majority of carcinomas-86% and 79% respectively. Expression of tumour markers was not associated with any certain histological type of carcinoma but was more characteristic for moderately and well differentiated adenocarcinomas. hCG positive tumour staining was less frequent than CEA and SP1 (only 22% tumours) and was much less intensive (5-50% population of carcinomatous cells) in the tumours. The study showed correlation between increased serum CEA concentration and tumour expression of antigen. PMID- 9337539 TI - Activity and tissue localization of cathepsin D in non small cell lung cancer. AB - Activity and tissue localization of cathepsin D were examined in tumors deriving from 80 patients with non small cell lung cancer. Activity of the enzyme was higher in sediments and supernatants of tumors than in non invaded lung parenchyma. In all histological types of tumors cathepsin D activity in sediments was three times lower and in lung parenchyma five times lower than in supernatants. The immunohistochemical technique was used for enzyme localization. We observed seemingly the lack of correlation between activity of cathepsin D examined in tumors and immunohistochemical reaction intensity in neoplasm cells. Different numbers of macrophages and quantity of tumor stroma could explain this effect in examined histological types of cancer. Results of our explanations indicates for relations between cathepsin D activity versus histological type and degree of tumor differentiation. We did not observe correlation between cathepsin D activity versus lymph node metastases and clinical stages. PMID- 9337538 TI - Activity and tissue localization of cathepsin G in non small cell lung cancer. AB - Activity and tissue localization of cathepsin G were examined in tumors deriving from 73 patients with non small cell lung cancer. Activity of cathepsin G was highest in adenocarcinoma, lower in planoepitheliale cancer, the lowest in macrocellular cancer. In all histological types of tumors cathepsin G activity in supernatants was lower than in sediments. The enzyme immunohistochemically was localized in neutrophils. There is evident correlation between neutrophil numbers and cathepsin G activity in examined cancer types. Result of our examinations indicate a relationship between cathepsin G activity, grade of tumor differentiation and particular clinical stages of disease. PMID- 9337540 TI - Prolidase and prolinase activities in moderately and poorly differentiated lung adenocarcinoma. AB - Prolidase (E.C.3.4.13.9) and prolinase (E.C.3.4.13.8) activities were determined in normal human lung and human lung adenocarcinomas of various degree of histologic differentiation. Since these dipeptidases were found to be enzymes catabolizing mainly collagen and simultaneously involved in the recycling of proline for collagen biosynthesis, the measurement of this protein and its degradation products in studied tissues (by hydroxyproline determination) was performed. It has been found that the activity of prolinase in G2-moderately and G3-poorly differentiated lung adenocarcinoma groups, was elevated compared to lung parenchyma and that the increase was proportional to the degree of adenocarcinoma differentiation. Prolidase activity was elevated only in G3 lung adenocarcinoma. The increase of prolidase and prolinase activities were accompanied by an increase of tissue collagen content. Collagen degradation products (CDP) represented one third of total collagen in control lung tissues while in lung adenocarcinomas the CDP represented significantly lower percentage of total tissue collagen. The results suggest that prolidase and prolinase activities may reflect: (i) degree of differentiation of lung adenocarcinoma and (ii) disturbances in tissue collagen metabolism. PMID- 9337541 TI - Prolidase activity and beta 1 integrin expression in moderately and poorly differentiated lung adenocarcinomas. AB - Primary human lung adenocarcinomas were divided into two groups according to the degree of histologic differentiation: G2-moderately and G3-poorly differentiated tumors. Each group was compared with normal lung tissue in respect to prolidase activity, its ability to interact with specific antibody, free proline and beta 1 integrin subunit content as well as ability of beta 1 integrin subunit to interact with specific antibody. It was found that prolidase activity in lung adenocarcinomas G3, was significantly elevated in comparison to normal lung tissue. In lung adenocarcinoma G2 no significant changes in the enzyme activity were observed. Increase in the enzyme activity was accompanied by increase of free proline content in the tissues. The western blot analysis revealed that prolidase of lung adenocarcinomas is identical to prolidase originated in control lung tissue. It was noticed that elevated activity of prolidase in adenocarcinomas G3 was accompanied by its high expression. In respect to beta 1 integrin expression, known to play an important role in metastasis, no difference was found between adenocarcinoma groups and the control lung tissue. The presented data suggest that the level of prolidase activity in lung adenocarcinoma may serve as a more sensitive marker for the histologic degree of malignancy, than the level of beta 1 integrin expression. PMID- 9337542 TI - Cancer procoagulant (CP) in lung cancer. AB - Lung cancers (squamous cell carcinoma, microcellular carcinoma, macrocellular carcinoma and adenocarcinoma) show procoagulant activity. It mainly depends on the presence of cancer procoagulant (CP) in lung cancer cells. PMID- 9337543 TI - Studies on angiogenesis intensity in lung cancer in aspect of its correlation with histological type of tumor and clinical stage. AB - Angiogenesis intensity in lung cancer, in compliance with histological types, tumor differentiation and different clinical stage of disease, was evaluated. The group of 65 patients, 34-73 years old (average 58), who have been operated, were examined. Microvessels were highlighted by immunohistochemical method staining of endothelial cells for factor VIII-von Willebrand. Microvessel and single endothelial cell count per 1 mm2 in each section was determined using light microscope, synchronized with camera and IBM-AT computer (LUCIA-NIKON program for morphometric studies). All cases were divided into three groups depending on angiogenesis intensity: Io-0-200, IIo-201-400, IIIo-400 angiogenic objects/mm2 (microvessels-MV plus endothelial cells-EC). Majority (57%) of examined cases were found in IIo group. The results of studies on angiogenic objects number (MV+EC) per 1 mm2 in different histological type of cancer were following: 248.97 +/- 114.72 in squamous cell, 253.18 +/- 81.32 in adenocarcinoma, 284.04 +/- 114.27 in large cell, 388.02 +/- 117.73 in small cell, 385.27 +/- 210.92 in combined cancer. In each group of lung cancer with different TNM and clinical stages was found that the angiogenic objects number depends on T tumor feature, mainly in EC count analysis (T1-148.61 +/- 113.21, T2-179.38 +/- 100.57, T3 199.52 +/- 137.70, T4-253.18 +/- 108.60). Obtained data were analyzed with of t Student's test. The differences between angiogenic objects number in the groups with different histological type of lung cancer were no statistically significant, although were near threshold value in pairs squamous cell versus small cell (p = 0.0545) and adenocarcinoma versus small cell (p = 0.0611). The differences of EC counts in the same pairs were statistically significant: p = 0.0247 (squamous cell versus small cell) and p = 0.0380 (adenocarcinoma versus small cell). The correlation between angiogenic objects number and grade of tumor differentiation was statistically significant for G1 group versus G2 (p = 0.0380) and G1 versus G4 (p = 0.0008), in comparison to G2 versus G4-p = 0.0688. The remaining results were not statistically significant. Obtained data are no final because the examined groups of cases were not numerous enough. The dependences should be examined in large series of cases. PMID- 9337545 TI - Ultrasound vascular imaging with subtraction in evaluation of lung tumor vascularity--a case report. AB - Ultrasound vascular imaging with subtraction is considered as very useful method to evaluation vascularity of tumor and its relationships to great vessels. Lung tumor was evaluated with power color Doppler ultrasound with and without B-mode scanning. Power color Doppler imaging without B-mode scanning is better depicted tumor internal vascular architecture and its relationships to great vessels then with B-mode. This method makes differentiation of tumor character possibly and interventional procedures more safely. PMID- 9337544 TI - Immunohistochemical evaluation of tumour angiogenesis in adenocarcinoma and squamous cell carcinoma of lung. AB - Experimental studies revealed that growth and expansion of solid tumours depend on angiogenesis. Angiogenesis is very important factor for neoplastic metastasis. The presence of the metastasis is an ominous prognostic factor for many tumours, also for lung cancer. Studies of tumour microvessel density in resected non-small lung cancers have not given convincing data about value of angiogenesis. Only few reports regarded the association with angiogenesis in different histological types in lung carcinoma. Samples of 35 adenocarcinomas and 41 squamous cell resected, primary lung carcinomas were studied. Paraffin sections of tumours were stained immunohistochemically by antibody against endothelial marker CD34. Angiogenesis intensity was measured in the areas of the most active fields of tumour neovascularization. Microvessel density (MD) was higher in adenocarcinoma comparing to squamous cell cancer, but the difference was not statistically significant (p = 0,095). The groups of various stage of extension of disease in each histological type were compared-MD correlated with lymph node metastasis (p = 0,003) in the adenocarcinoma, whilst in squamous cell can cer differences between various groups of nodal involvement were not statistically significant (p = 0,53 and p = 0,22 respectively). Our results suggest that more intensive angiogenesis in adenocarcinoma could be more important factor for metastasis of adenocarcinoma than for squamous tumours. In the latter group angiogenesis may be more important for growth of squamous cell cancers, while the spread of squamous tumours may depend on other mechanisms. PMID- 9337546 TI - Screening of angiogenesis inhibitors by modified tumor-induced angiogenesis (TIA) test in lung cancer. AB - Aberrant angiogenesis-the new vessels formation is a mandatory event in the process of tumor growth and expansion. Studies on mechanisms involved in tumor induced angiogenesis (TIA) and on its possible inhibitors are needed in order to introduce in future new methods of tumor treatment. The aim of our study was to determine the usefulness of the modified cutaneous TIA (mice without immunosuppression) test for screening in vivo of the angiogenesis modifiers. In both models (classical and modified TIA) we demonstrated comparable angiogenesis activity following human lung cells inoculation and similar degree of neovascularization response inhibition caused by theobromine. We reported that in modified TIA model preincubation with theobromine significantly suppressed angiogenic potential of human lung cancer cells as well as the ability of those cells to produce proangiogenic cytokine-bFGF. PMID- 9337547 TI - Antipyrine metabolism in lung cancer patients and their relatives. AB - The existence of large interindividual differences in oxidation phenotypes and link of them with susceptibility to certain cancers was shown in animal and human studies. In a new approach plasma antipyrine half-lives have been measured in 60 lung cancer patients, 56 their first degree relatives and 75 healthy controls without cancer in their families. The mean antipyrine half-lives were significantly shorter (p < 0.005) in lung cancer patients group and in the group of their relatives when compared with the cancer free matched control group. This difference remained significant after adjusting for smoking gender and age. Six adenocarcinoma patients have shown significantly faster antipyrine metabolism than subjects with other histological types of lung cancer and obviously than healthy control. PMID- 9337548 TI - Fine needle aspiration biopsy cytology of pulmonary tumors. AB - Transthoracic fine needle aspiration specimens of the pulmonary tumors were obtained from 144 patients. Fine needle aspiration biopsy were performed using local anaesthesia and ultrasonographic or scopie control. The 20-gauge needles were used to obtain the specimens. 93 of patients have been diagnosed as squamous cell carcinomas, 28 as adenoid carcinomas, 18 as anaplastic carcinomas and 5 as a non neoplastic lesions (tuberculoma and abscessus). The tumors were typed according to the second WHO histological classification. Analysis of the date indicated that malignant neoplasms were identified correctly with an accuracy of 92.0%. There were not false positive diagnoses. There were two false negative diagnoses (squamous cell carcinoma) and in three cases the diagnosis were as suspected for malignancy. The results confirmed the value of fine needle aspiration cytopathology for the diagnosis of pulmonary tumors. PMID- 9337549 TI - Evaluation of colour Doppler sonography in lung tumor biopsy. AB - Between 1986 and 1995, 1800 ultrasound examinations of the chest were done. Addition of colour Doppler imaging improves safety of procedures and allowed to perform ultrasound guided biopsy in 47 patients. On the basis of our experience we state that colour Doppler sonography guided needle aspiration biopsy is useful, sufficient and safety diagnostic method of malignant lung masses, especially peripheral and wall-chest located. PMID- 9337550 TI - The management of tracheal obstruction. AB - The stenting of the tracheobronchial tree has become common method of palliative treatment in inoperable cases. The patients with malignant tracheal stenosis constitute the most numerous group. The airway patency should be restored prior to inserting the stent. The stent protects both trachea and bronchi from narrowing their lumen with malignant tissue. When malignant infiltration is accompanied by airway compression the stenting is the method of choice. The own model of tracheobronchial prosthesis has been invented. It enables the protection of both trachea and the bronchi. The stent is inserted endoscopically on rigid bronchoscope. It is the method of stenting, which allows the visual control during the whole procedure. PMID- 9337551 TI - Electrokinetic potential of lung cancer cells. AB - Charge density and electrokinetic potential have been determined of the cells obtained from unchanged lung parenchyma tissue, from bronchus epithelium and from lung cancer. The measurements were carried out in the physiological solution by the electrophoretic method. The tissues were separated into cells using trypsin. The electrokinetic potential of the cancer cells was found to be shifted to lower potentials with respect to unchanged lung tissue cells, i.e. to have dropped by about 20%. The estimate was done with 26 cases. PMID- 9337552 TI - Protective effect of human recombinant tumour necrosis factor-alpha against lung metastases of the Morris hepatoma in rats. AB - The effect of intratumour (i.t.) administration of human recombinant tumour necrosis factor-alpha (hrecTNF-alpha) on Morris 5123 hepatoma spontaneous lung metastases was studied. Tumour was implanted in the skeletal muscles of the right hind limb of Buffalo rats. Two weeks after the implantation treatment with cytokine started. The cytokine was injected i.t. in a single dose of 10 micrograms in 4- and 8-day cycles. In control animals PBS was administered, respectively. Then all rats were necropsied and the extent of lung metastases was determined. Data were expressed as volume of lung metastases. In this study we observed significant antimetastatic effects of hrecTNF-alpha (p < 0.05) determined by reduced volume of lung metastases in treated animals. PMID- 9337553 TI - Alveolar epithelial cells after intratumour administration of HREC TNF-alpha mutein VI. AB - The aim of this study was to explore the effect of intratumour mutein VI-Hrec TNF alpha administration upon the ultrastructural changes within the pulmonary tissue, with special attention paid to type II alveolar epithelial cells. The experiment was carried out on Buffalo rats with implantable Morris hepatoma series 5123 in the skeletal muscles of the limb. Mutein VI-Hrec TNF-alpha was administered in a dose of 10 micrograms once a day in a cycle of eight days. Control animals were given saline (PBS). Ultrastructural changes within the pulmonary tissue were evaluated in the electron transmission microscope (TEM), with special attention paid to alveolar epithelial cells. In the animals receiving hrec TNF-alpha mutein VI, damage to the alveolar epithelial cells was found. In the later period (14 days after the mutein treatment), repairing processes were observed, accompanied by intensified fibrotic processes in the interalveolar septal interstitium, with the subsequent pulmonary tissue rebuilding. The study confirmed the possibility of a peripheral action of hrec TNF-alpha mutein VI after its administration to the experimental Morris hepatoma and found the alveolar epithelial cells to be a key element of the pulmonary tissue subjected to the cytokine effect. PMID- 9337554 TI - Comparative studies on the effect of TNF-alpha and endotoxin on the ultrastructural picture of pulmonary capillaries in pregnant rabbits. AB - The aim of the present study was the comparative analysis of morphological changes found in the pulmonary microcirculation of pregnant rabbits in the course of experimental septic shock induced by endotoxin or human recombinant TNF-alpha administration. The experiments used 30 female rabbits, white Dutch, c.3 kg mean body weight. The endotoxin Escherichia coli, serotype S.0127: 138 (Sigma) was applied in a single dose of 100 micrograms/kg b.w. intraperitoneally. The human recombinant TNF-alpha (biological activity 2-4 x 10(7) U/mg of protein) was injected intraperitoneally, also once, in a dose of 100 micrograms/kg b.w. Morphological examinations were based on the ultrastructural analysis in the transmission electron microscope. The ultrastructural analysis revealed no morphological differences between pregnant and non-pregnant rabbits which received TNF-alpha. However, significant differences were observed between animals given TNF-alpha and those given the endotoxin. These differences referred to endothelium damage degree and to the cellular composition of inflammatory infiltrations. More severe damage to endothelial cells (necrosis inclusive) was observed in the endotoxin-treated rabbits. Blood vascular lumen in these animals showed cellular aggregation, of neutrophils and platelets in particular as well as microthrombi. An increased tendency towards the development of these changes was noted in pregnant rabbits. The lumen of pulmonary capillaries in TNF-alpha treated animals showed domination of monocytic cells. Features of endothelial cell stimulation were observed, although without a tendency to form microthrombi. PMID- 9337556 TI - The ethanol effect on lung metastases development in experimental Morris 5123 hepatoma. AB - The experiment was carried out on 45 male Wistar rats which were divided into 3 groups: group I-15 rats which were given water to drink during the experiment, group II-15 rats obtaining 10% ethanol solution during the experiment and group III-15 rats obtaining 20% ethanol solution during the experiment. All animals were injected 0.3 ml suspension of hepatoma Morris 5123 cells directly to the liver on the 14th day of the experiment. After the 9 weeks of the experiment the animals from all groups were narcotized, decapitated and the lungs were taken into the morphometric, histological and ultrastructural examinations. They showed that ethanol has a stimulating effect on formation and development of hepatoma Morris 5123 in rat lungs. The increase of number and extensiveness of metastases as well as the increase of mean metastases focuses volume, and the increase of lung weight in animals which obtained ethanol are the exponent of the influence. On the basis of ultrastructural examinations, it can be noted that promoting activity of ethanol can be connected: a) with the increase of neoplastic cells activity, b) with changes appearing in pulmonic epithelium and endothelium of vessel enabling neoplastic cells to adhere to basal membrane as well as their crossing the vessel wall. PMID- 9337555 TI - Quantitative and morphological changes in BAL-isolated cells after the administration of TNF-alpha into Morris hepatoma. AB - The aim of this study was to explore the effect of intratumor TNF-alpha administration upon the composition and adherence degree of cells isolated from the lungs through multiple bronchoalveolar lavages (BAL). Ultrastructural evaluation of BAL-isolated cells was performed in the scanning electron microscope (SEM). The experiment used Buffalo rats. A suspension of 3 x 10(6) cells of Morris hepatoma (5123 series) was injected to the right hind leg of the animals. After fourteen days, TNF-alpha was administered into the tumor in a dose of 1.5 x 10(4) U in 0.5 ml PBS solution. The animals of group I were given 4 doses of TNF-alpha and group II-8 doses of TNF-alpha every 24 hours. Control groups consisted of rats with injected Morris hepatoma which were given PBS solution instead of TNF-alpha (group III A, B) and animals without the hepatoma, given intramuscullary 4 or 8 TNF-alpha, respectively (groups IV A, B). No statistically significant differences were noticed between groups I and II with regard to the number of macrophages and neutrophils isolated from the rat lungs compared with control group IV. However, such differences were observed compared with group III. In group II and IV B, an increase in the adherence of isolated cells was found compared with group III, as well as arise in the number of macrophages with the largest diameters. We found a correlation between the increase in cell adherence and ultrastructural changes (in SEM) suggesting an increased activity of BAL-isolated cells. PMID- 9337557 TI - Contribution of capillary endothelium to the processes of pulmonary tissue rebuilding in the course of acute enzymatic lung injury. AB - Ultrastructural analysis was made of the pulmonary capillary endothelial cells, with special regard paid to their possible contribution to the processes of pulmonary tissue rebuilding initiated by intratracheal infusion of a proteolytic enzyme-papain. Experimental animals (male Wistar rats) were additionally given BCG vaccine in doses activating the alveolar macrophage system. The study was performed after 1 day, and 1, 4 and 12 weeks following papain administration. In the animals given BCG vaccine, ultrastructural exponents of endothelial cell activation were found. Cumulation of inflammatory cells, mainly monocytes, was observed in vascular lumen. Application of papain solution to animals caused a number of damaging and exudative changes, being most pronounced within vascular endothelium in animals simultaneously given BCG vaccine. In later periods, rebuilding and productive processes, accompanied by collagen cumulation in the interstitium of interalveolar septa, dominated. In the animals receiving BCG vaccine and papain, ultrastructural pictures indicated active contribution of endothelial cells to the processes of pulmonary tissue rebuilding initiated by intratracheal infusion of the proteolytic enzyme. PMID- 9337558 TI - Ultrastructural analysis of the pneumocyte-interstitium boundary line in the course of enzymatic lung injury. AB - The studies were performed basing on the experimental model of acute pulmonary tissue injury. Papain in a dose of 2 mg/ml PBS/100 g b.w. was administered once, intratracheally, on the 21st day of the experiment. Besides, female Wistar rats were injected twice with BCG vaccine in a dose of 4 x 10(8) microorganisms. BCG vaccine was administered intraperitoneally on the 1st and 14th day of the experiment to activate the system of mononuclear phagocytes. Control rats were intratracheally or/and intraperitoneally given PBS solution. All the animals were killed on the 28th, 35th and 42nd day of the experiment. A single intratracheal papain injection induced emphysematous changes in the animal lungs. The changes were accompanied by basement membrane rebuilding and focal collagen and elastin cumulation. An increase in the number of type II alveolar epithelial cells was observed. Anchorage of collagen fibres and microfibrillary structures in the cytoplasm of type II pneumocytes was observed in the BCG- and papain-treated animals. There, the cytoplasmic membrane of type II cells was completely indistinct and the cytoplasm formed processes to penetrate into the connective tissue fibres. The results obtained indicate possible contribution of type II pneumocytes to fibroplasia processes during lung parenchyma rebuilding and suggest the necessity to include fibroplasia elements in the existing definition of emphysema. PMID- 9337559 TI - Evaluation of the effect of macrophage system activation on the intensity degree of early destructive changes in acute enzymatic lung injury. AB - The effect of macrophage system activation on the intensity degree of early destructive changes induced with intratracheal infusion of a proteolytic enzyme papain was evaluated in the paper. The study was carried out on male Wistar rats. BCG vaccine was used as a macrophage system activating substance. Damage degree of the pulmonary tissue was evaluated after 2 hours following papain administration by determining W/D index (wet lungs/dry lungs). Spectrophometric analysis of bronchoalveolar lavages (BAL) was made in order to evaluate the advancement of haemorrhagic alterations in the lungs. Damage to the pulmonary tissue was greater in the animals given BCG vaccine and papain, compared with controls and papain treated animals. Differences in the intensity degree of destructive changes assessed using both the W/D index and haemoglobin content in BAL were statistically significant. The results obtained indicate a particular role of activation of the macrophage system in the development of early destructive changes due to intratracheal administration of proteolytic enzymes and point to the significance of these changes in the morphogenesis of experimental lung emphysema. PMID- 9337560 TI - Cervical cancer and human papillomavirus: epidemiological evidence and perspectives for prevention. AB - Cervical cancer is a major public health problem, as it is the second most common cancer in women world-wide after breast cancer. About 80% of the half a million cases estimated to occur annually in the world, occur in developing countries. The epidemiological evidence linking human papillomavirus (HPV) to cervical cancer is reviewed. It is concluded that over 90% of cervical cancers can be attributed to certain HPV types. HPV 16 accounts for the highest proportion (50%) followed by HPV 18 (12%), HPV 45 (8%) and HPV 31 (5%). The association with these HPV types are very strong and consistent with odds ratios over 15 in all case control studies in high- and low-risk countries for cervical cancer. However, HPV is not a sufficient cause of this malignancy; certain cofactors are necessary for a proportion of HPV persistent infections to eventually progress to cancer. These include host factors such as histocompatibility types and immunological response, hormonal influences and infections with other sexually transmitted agents such as Chlamydia trachomatis. In addition, results from our studies carried out in Spain and Colombia support the hypothesis that male carriers of HPV play an important role in the development of cervical cancer in their wives. The recognition of the central role of HPV in cervical cancer has far-reaching implications for the primary and secondary prevention of this malignancy. Prophylactic and therapeutic HPV vaccines are now under development and HPV typing is being integrated into screening programmes in pilot studies in a few developed countries. In developing countries, well conducted conventional screening programmes remain the best approach for the control of cervical cancer until a safe and efficient HPV vaccine can be used in the general population. PMID- 9337561 TI - Preventive vaccines for cervical cancer. AB - The potential use of vaccines for the human papillomavirus (HPV) in the prevention and treatment of cervical cancer is a possibility in the near future. Close to 20 genotypes of HPV, of the 75 that have been identified, infect the feminine genital tract, but four subtypes (16, 18, 31 and 45) have been associated in close to 80% of cervical cancers, this article proposes that in order to design an effective prophylactic vaccine against HPV infection, an adequate immune response should be guaranteed through four goals; a) activation of antigens present in the cell; b) overcoming the host response and viral genetic variability in the T cell response; c) generation of high levels of T and B memory cells; and d) persistence of antigens. PMID- 9337562 TI - Nutrition and cancer. AB - Evidence from both animal and epidemiologic studies indicate that throughout life excessive energy intake in relation to requirements increases risk of human cancer. Rapid growth rates in childhood lead to earlier age at menarche, which in turn increases risk of breast cancer, and accumulation of body fat in adulthood in related to cancers of the colon, kidney, and endometrium as well as postmenopausal breast cancer. Higher intake of vegetables and fruits has been associated with lower risks of many cancers. The constituents responsible for these apparent protective effects remain uncertain, although evidence supports a contribution of folic acid. Recent evidence suggests that the percentage of energy from fat in the diet is not a major cause of cancers of the breast or colon. Higher intake of meat and dairy products has been associated with greater risk of prostate cancer, which may be related to their saturated fat content. Also, red meat consumption has been associated with risk of colon cancer in numerous studies, but this appears to be unrelated to its fat content. Excessive consumption of alcohol increases risks of upper gastrointestinal tract and even moderate intake appears to increase cancers of the breast and large bowel. Although many details remain to be learned, evidence is strong that remaining physically active and lean throughout life, consuming an abundance of fruits and vegetables, and avoiding high intakes of red meat, foods high in animal fat, and excessive alcohol will substantially reduce risk of human cancer. PMID- 9337563 TI - The role of chemoprevention in cancer control. AB - Chemoprevention can be defined as the use of chemical compounds or medicines to prevent the occurrence of precancerous lesions (markers) or to slow down or revert the progression of clinically established disease. The use of randomized trial design is considered the gold standard for evaluating the preventive value of chemicals against cancer, since they control for confounding and avoid information bias. The principal school in relation to cancer control through chemoprevention is based on studies of cancer and diet. Initially, ecological studies set the cornerstone, but later case-control studies supported the hypothesis of an inverse association between foods and cancer risk (principally epithelial), suggesting that determined micronutrients participate as protection in this process. Other studies include specific chemical analyses, which have potential problems that could lead to erroneous conclusions, such as sample and measurement errors. During this decade randomized intervention trials have been carried out to test this hypothesis, but conclusions have been so diverse and the designs used have been so different in terms of levels of exposure, that consistent conclusions are not possible. We can conclude that using studies with randomized, double-blind, controlled designs is interesting, but problems remain to be solved, including: agent selection, the design to be chosen, and especially the balance between benefits sought and secondary effects, including cost effectiveness, since some chemicals cannot compete with other preventive or therapeutic measures. PMID- 9337564 TI - Epidemiology of gastric cancer and perspectives for prevention. AB - The most recent estimates of the world-wide incidence of cancer indicate that gastric cancer was in 1990 the second most frequent cancer in the world (after lung cancer), with about 900000 new cases diagnosed every year. Steady declines in the rates have been observed everywhere in the last few decades, but the absolute number of new cases per year is increasing mainly because of ageing of the population. The exact causes of the decline of gastric cancer are not well understood, but must include improvements in diet, food storage (e.g., refrigeration) and, possibly, the decline of Helicobacter pylori infection. Dietary modifications and, possibly, vitamin supplements remain one of the most important tool for the prevention of gastric cancer. Control of H. pylori infection, by means of eradication or immunization, is also likely to offer great potential for the prevention of this important malignancy. PMID- 9337565 TI - Tobacco smoking and cancer: the promise of molecular epidemiology. AB - Neoplastic development is a multistage process that includes multiple genetic changes. In this article the authors review studies on molecular epidemiology of tobacco. Current concepts of the multistage carcinogenic model are reviewed, as are their use in observational studies. Finally, benzo[a]pyrene are analyzed as an example. PMID- 9337566 TI - Ambient air pollution as a risk factor for lung cancer. AB - Epidemiologic studies over the last 40 years have observed that general ambient air pollution, chiefly due to the by-products of the incomplete combustion of fossil fuels, is associated with small relative increases in lung cancer. The evidence derives from studies of lung cancer trends, studies of occupational groups, comparisons of urban and rural populations, and case-control and cohort studies using diverse exposure metrics. Recent prospective cohort studies observed 30-50% increases in the risk of lung cancer in relation to approximately a doubling of respirable particle exposure. While these data reflect the effects of exposures in past decades, and despite some progress in reducing air pollution, large numbers of people in the US continue to be exposed to pollutant mixtures containing known or suspected carcinogens. These observations suggest that the most widely cited estimates of the proportional contribution of air pollution to lung cancer occurrence in the US, based largely on the results of animal experimentation, may be too low. It is important that better epidemiologic research be conducted to allow improved estimates of lung cancer risk from air pollution in the general population. The development and application of new epidemiologic methods, particularly the improved characterization of population wide exposure to mixtures of air pollutants and the improved design of ecologic studies, could improve our ability to measure accurately the magnitude of excess cancer related to air pollution. PMID- 9337567 TI - Cancer epidemiology and the workplace. AB - Occupational exposure occurs most frequently through direct contact with carcinogenic agents, with any of their active metabolites during absorption (skin, respiratory tract); or during excretion (urinary tract). In the USA, from 2-8% of cancer are attributed to this circumstance. In developing countries emphasis should be made on prevention measures of possible carcinogenic exposure factors, with three basic premises: a) identify exposure markers (biological monitoring); b) identification of high risk subjects, presumable before exposure occurs, and c) early identification of signs of sickness (medical surveillance). This article proposes that, in theory, all occupational cancer can be prevented. PMID- 9337569 TI - The rehabilitation dream. PMID- 9337570 TI - Characteristics of alleged malpractice. PMID- 9337568 TI - Estrogens and breast cancer. AB - In this review, we summarize the epidemiologic evidence for the associations of oral contraceptives and postmenopausal hormones with risk of breast cancer. We also describe the biologic plausibility of these relationships. Overall, there appears to be little, if any, increase in risk with oral contraceptive use in general, even among users for 10 or more years. However, compared to never users, current oral contraceptive users appear to have a modest elevation in risk that subsides within about 10 years after cessation of use. For postmenopausal hormones, the weight of the evidence suggests little or no increase in risk among users of short duration, or for use in the past. However, current longer term use is associated with an increased risk of breast cancer that increases with duration. This increase in risk is large enough, and well enough supported, to be considered along with the other risks and benefits of postmenopausal hormone therapy. PMID- 9337571 TI - Pain management. PMID- 9337572 TI - Developing a compliance program to identify and prevent inaccurate or improper Medicare billing practices. PMID- 9337573 TI - Deferred demise. PMID- 9337574 TI - Post-traumatic regeneration, neurogenesis and neuronal migration in the adult mammalian brain. AB - Unlike the peripheral nervous system (PNS), the mammalian central nervous system (CNS) clearly lacks the robust regenerative characteristics and capacity of the former. Despite this fact, two unique regions of the adult mammalian CNS possess such regenerative potential and are capable of active regeneration following injury or structural compromise. These unique areas are the olfactory system and the neurohypophyseal system of the endocrine hypothalamus. Furthermore, it has been clearly demonstrated that primordial neuroblasts regarded as stem cells emerge from the subependymal parenchyma of the walls and floor of the third cerebral ventricle, migrate to the ventricular surface and undergo compensatory synaptogenesis within one week following hypophysectomy. In situ hybridization studies have unequivocally demonstrated that the up-regulation of nitric oxide synthase (NOS) is essential for neural (axonal) regeneration and neuronal (stem cell) migration to occur. Moreover, neuronal migration is reliably inhibited following the administration of the NO antagonist, nitroarginine. The current investigation serves to confirm a remarkable degree of plasticity and regeneration in the adult mammalian neurohypophyseal system coupled with the emergence of primordial neuroblasts that undergo apparent differentiation, migration and compensatory synaptogenesis in response to the up-regulation of NO that occurs following the trauma of hypophysectomy. Evidence from the current investigation appears to confirm that specialized glia of the neurohypophyseal system, the so-called pituicyte, proliferate following hypophysectomy and may serve as a growth matrix or structural template that may target and direct regenerating Supraoptic (SON) and Paraventricular (PVN) axons toward endothelial primordia in the regenerating neural stem and lobe. PMID- 9337576 TI - The computerized patient record: managing clinical data. PMID- 9337575 TI - Memoir of Nina B. Woodside 1931-1977. PMID- 9337577 TI - History of modern medicine. PMID- 9337578 TI - Features and their configuration in face recognition. AB - Tanaka and Farah (1993) have proposed a holistic approach to face recognition in which information about the features of a face and their configuration are combined together in the face representation. An implication of the holistic hypothesis is that alterations in facial configuration should interfere with retrieval of features. In four experiments, the effect of configuration on feature recognition was investigated by creating two configurations of a face, one with eyes close together and one with eyes far apart. After subjects studied faces presented in one of the two configurations (eyes-close or eyes-far), they were tested for their recognition of features shown in isolation, in a new face configuration, and in the old face configuration. It was found that subjects recognized features best when presented in the old configuration, next best in the new configuration, and poorest in isolation. Moreover, subjects were not sensitive to configural information in inverted faces (Experiment 2) or nonface stimuli (i.e., houses; Experiments 3 and 4). Importantly, for normal faces, altering the spatial location of the eyes not only impaired subjects' recognition of the eye features but also impaired their recognition of the nose and mouth features-features whose spatial locations were not directly altered. These findings emphasize the interdependency of featural and configural information in a holistic face representation. PMID- 9337579 TI - Encoding, repetition, and the mirror effect in recognition memory: symmetry in motion. AB - Attention/likelihood theory has been used to explain the mirror effect in recognition memory. The theory also predicts that any manipulation that affects the recognition of old items will also affect recognition of the new items-more specifically, that all the underlying distributions will move and that they will move symmetrically on the decision axis. In five experiments, we tested this prediction. The first two experiments used encoding tasks during study to change recognition performance for high- and low-frequency words. The results show symmetrical dispersion of the underlying distributions. The final three experiments used repetition to increase recognition performance. Repetition produced a symmetrical pattern of movement that was different from that produced by encoding task. This pattern is, however, also covered by attention/likelihood theory. A further extension of the theory was used to predict response times. PMID- 9337581 TI - Associative and similarity-based processes in categorization decisions. AB - Two experiments were directed at distinguishing associative and similarity-based accounts of systematic differences in categorization time for different items in natural categories. Experiment 1 investigated the correlation of categorization time with three measures of instance centrality in a category. Production frequency (PF), rated typicality, and familiarity from category norms for British participants (Hampton & Gardiner, 1983) were used to predict mean categorization times for 531 words in 12 semantic categories. PF and typicality (but not familarity) were found to make significant and independent contributions to categorization time. Error rates were related only to typicality (apart from errors made to ambiguous or unknown items). Experiment 2 provided a further dissociation of PF and typicality. Manipulating the difficulty of the task through the relatedness of the false items interacted primarily with the effect of typicality on categorization time, whereas, under conditions of easy discrimination, prior exposure to the category exemplars affected only the contribution of PF to the decision time. The dissociation of typicality and PF measures is interpreted as providing evidence that speeded categorization involves both retrieval of associations indexed by PF and a similarity-based decision process indexed by typicality. PMID- 9337580 TI - Categorizing chairs and naming pears: category differences in object processing as a function of task and priming. AB - Four experiments are reported examining the locus of structural similarity effects in picture recognition and naming with normal subjects. Subjects carried out superordinate categorization and naming tasks with picture and word forms of clothing, furniture, fruit, and vegetable exemplars. The main findings were as follows: (1) Responses to pictures of fruit and vegetables ("structurally similar" objects) were slowed relative to pictures of clothing and furniture ("structurally dissimilar" objects). This structural similarity difference was greater for picture naming than for superordinate categorization of pictures. (2) Structural similarity effects in picture naming were reduced by repetition priming. Repetition priming effects were equivalent from picture and word naming as prime tasks. (3) However, superordinate categorization of the prime did not produce the structural similarity effects on priming found for picture naming. Furthermore, such priming effects did not arise for picture or word categorization or for reading picture names as target tasks. It is proposed that structural similarity effects on priming object processing are located in processes mapping semantic representations of pictures to name representations required to select names for objects. Visually based competition between fruit and vegetables produces competition in name selection, which is reduced by priming the mappings between semantic and name representations. PMID- 9337582 TI - Location errors in partial-report bar-probe experiments: in search of the origin of cue-alignment problems. AB - In studies using Averbach and Coriell's (1961) partial-report bar-probe paradigm with linear arrays, most errors involve the naming of an item that was in the display but in a position other than the cued one. Up to now, there is no general agreement on the origin of these location errors. Point of departure in this paper is that part of the location errors arises from inappropriate application of the cue. It is tested whether this originates from problems to perceive the position of the cue ("cue-displacement hypothesis") or from confusion about the order of the items in the array ("item-order hypothesis"). The results of two bar probe experiments are reported. A novel, crucial, finding in both experiments is that, among the location errors, there was a preponderance of response letters that came from the central side of the cued item. In the second experiment, this was observed not only in the usual postcue conditions but also when the cue preceded the array. These results positively corroborate the cue-displacement hypothesis and do not support the item-order hypothesis: The cue tends to be perceived more toward the center of the visual field than it actually is exposed that is, there is a central drift of the cue. PMID- 9337584 TI - At-lexical, articulatory interference in silent reading: the "upstream" tongue twister effect. AB - In two experiments, we investigated the interpretation and boundary conditions of the tongue-twister (TT) effect in silent reading. Previously, McCutchen, Bell, France, and Perfetti (1991) observed a TT effect when students made semantic acceptability judgments on sentences, but not when they made lexical decisions on lists of words. Using similar methodology in Experiment 1, along with two changes (using "better" TTs and longer word lists), we observed a TT effect in a lexical decision task. In Experiment 2, a memory span task revealed that students recalled fewer words from TT lists than from control lists. These results suggest that the basic mechanism of the TT effect may be articulatory, rather than working-memory, interference that occurs during lexical access and resurfaces post-lexically, inhibiting efforts to maintain the temporal order of several words. PMID- 9337583 TI - Linguistic focus affects eye movements during reading. AB - In two experiments, we explored how readers encode information that is linguistically focused. Subjects read sentences in which a word or phrase was focused by a syntactic manipulation (Experiment 1) or by a preceding context (Experiment 2) while their eye movements were monitored. Readers had longer reading times while reading a region of the sentence that was focused than when the same region was not focused. The results suggest that readers encode focused information more carefully, either upon first encountering it or during a second pass reading of it. We conclude that the enhanced memory representations for focused information found in previous studies may be due in part to differences in reading patterns for focused information. PMID- 9337585 TI - The influence of word function in the missing-letter effect: further evidence from French. AB - When asked to detect target letters while reading continuous text, subjects miss more letters in highly common function words than in less common content words. This is known as the missing-letter effect. According to the structural account, the higher omission rates for frequent function words are attributable to their role in supporting the extraction of phrase structure, after which they become lost in the transition from structure to meaning. This implies that word function in and of itself should affect letter detection accuracy. This issue was examined in four experiments while controlling for a number of confounded factors associated with another influential model: the unitization account. The first experiment extended the missing-letter effect to the French language. The second showed that letter detection is influenced by slight variations in the function assumed by the same word, such as when it is used as a definite article as opposed to a pronoun. This effect was observed even when the frequency of the orthographic pattern and the syllable stress patterns were controlled. In the last two experiments, a control was added for another factor: frequency of word meaning. The results indicate that word function contributes to the missing letter effect over and above what is contributed by frequency of word meaning. PMID- 9337586 TI - Egocentric spatial framework effects from single and multiple points of view. AB - In three experiments, we tested the one-place, one-perspective rule formulated by Franklin, Tversky, and Coon (1992). This rule proposes that subjects take a neutral, external perspective when they must use multiple viewpoints to make decisions about the locations of objects in memorized scenes. We compared responding from a single viewpoint with responding from two viewpoints. In Experiments 1 and 2, we used a sentence verification procedure, and in Experiment 3, we compared a true-false verification procedure with a six-alternative forced choice procedure. Under these various conditions, we observed egocentric spatial framework effects in that above-below judgments were faster than front-back judgments and front-back judgments were faster than right-left judgments. When responding from two points of view in a single place, our subjects took multiple intrinsic perspectives rather than one neutral external perspective as proposed by the one-place, one-perspective rule. PMID- 9337587 TI - Age differences in the allocation of study time account for age differences in memory performance. AB - How aging affects the utilization of monitoring in the allocation of study time was investigated by having adults learn paired associates during multiple study test trials. During each trial, a subject paced the presentation of individual items and later judged the likelihood of recalling each item on the upcoming test; after all items had been studied and judged, recall occurred. For both age groups in Study 1, (1) people's judgments were highly accurate at predicting recall and (2) intraindividual correlations between judgments (or recall) on one trial, and study times on the next trial were negative, which suggests that subjects utilized monitoring to allocate study time. However, the magnitude of these correlations was less for older than for younger adults. Study 2 revealed that these differences were not due to age differences in forgetting. Results from both studies suggest that older adults do not utilize on-line monitoring to allocate study to the same degree as younger adults do, and that these differences in allocation contribute to age deficits in recall. PMID- 9337588 TI - Self-events and other-events: temporal dating and event memory. AB - A diary methodology was used to assess factors related to temporal dating and cued recall of real-world events. In one diary, participants kept a record of unique personal autobiographical events. In a second diary, participants recorded unique events from the life of a friend or relation. At the time each event was recorded, participants rated the event's pleasantness, person typicality, and degree of initial mental involvement in the event. At the end of the academic quarter, participants provided a recall rating, a rehearsal rating, a date estimate, and a report of the strategy used to estimate the date for each event. Results of regression analyses indicated that both self-events and other-events were characterized by superior memory for person-atypical events. Furthermore, there was a positivity bias in recall for self-events, but there was a negativity bias in recall for other-events. Mediational analyses indicated that the self event positivity bias was due to enhanced mental involvement when the events occurred, whereas the other-event negativity bias was due to subsequent event rehearsal. The date estimation results indicated that self-event dating was more accurate and evinced less telescoping than other-event dating. Furthermore, the accuracy of date estimates was substantially mediated by event memory. However, mediational differences between self-events and other-events did not emerge. The theoretical implications of these results are discussed. PMID- 9337589 TI - Rate of temporal discounting decreases with amount of reward. AB - The present, subjective value of a delayed reward is a decreasing function of the duration of the delay. This phenomenon is termed temporal discounting. To determine whether the amount of the reward influences the rate of temporal discounting, we had subjects choose between immediate and delayed hypothetical rewards of different amounts ($100, $2,000, $25,000, and $100,000 delayed rewards). As predicted by psychological models of the choice process, hyperbolic functions described the decrease in the subjective value of the delayed reward as the time until its receipt was increased (R2s from .86 to .99). Moreover, hyperbolic functions consistently provided more accurate descriptions of the data than did exponential functions predicted by an economic model of discounted utility. Rate of discounting decreased in a negatively accelerated fashion as the amount of the delayed reward increased, leveling off by approximately $25,000. These findings are interpreted in the context of different psychological models of choice, and implications for procedures to enhance self-control are discussed. PMID- 9337590 TI - Artifactual power curves in forgetting. AB - Recent studies of the mathematical relationship between time and forgetting suggest that it is a power function rather than an exponential function, a finding that has important theoretical consequences. Through computational analysis and reanalyses of published data, we demonstrate that arithmetic averaging of exponential curves can produce an artifactual power curve, particularly when there are large and systematic differences among the slopes of the component curves. A series of simulations showed that the amount of power artifact is small when the slopes of the component curves are normally or rectangularly distributed and when the performance measure is noise free. However, the simulations also showed that the artifact can be quite large, depending on the shape of the noise distribution and restrictions in the performance range. We conclude that claims concerning the form of memory functions should consider whether the data are likely to contain artifact caused by averaging or by the presence of range-restricted noise. PMID- 9337591 TI - Genuine power curves in forgetting: a quantitative analysis of individual subject forgetting functions. AB - Wixted and Ebbesen (1991) showed that forgetting functions produced by a variety of procedures are often well described by the power function, at-b, where a and b are free parameters. However, all of their analyses were based on data arithmetically averaged over subjects. R. B. Anderson and Tweney (1997) argue that the power law of forgetting may be an artifact of arithmetically averaging individual subject forgetting functions that are truly exponential in form and that geometric averaging would avoid this potential problem. We agree that researchers should always be cognizant of the possibility of averaging artifacts, but we also show that our conclusions about the form of forgetting remain unchanged (and goodness-of-fit statistics are scarcely affected by) whether arithmetic or geometric averaging is used. In addition, an analysis of individual subject forgetting functions shows that they, too, are described much better by a power function than by an exponential. PMID- 9337592 TI - Photo-cross-linked decyl methacrylate films for electrochemical and optical polyion probes. AB - Potentiometric and optical polyion probes based on photo-cross-linked thin films of decyl methacrylate (DMA) are described, and the effects of film composition on the response toward heparin are examined in detail. In accordance with existing theory governing potentiometric polyion response, lowering the amounts of plasticizer and tridodecylmethylammonium chloride ion exchanger within the film enhances its sensitivity toward heparin. Varying the cross-linker content of a DMA-based film, however, provides an additional mechanism to regulate its physical structure and, hence, the observed potentiometric polyion response. Films with low hexanedioldimethacrylate cross-linker content yield optimal potentiometric heparin detection limits (0.04 microM), suggesting a lower diffusion coefficient within such films, apparently due to interactions between adjacent pendant decyl groups. Increasing crosslinker content interrupts these interactions and facilitates diffusion. This knowledge is applied to optimize optical heparin sensing via DMA films covalently attached to glass substrates. When used in a limited volume/fixed exposure time measurement mode, such optically sensitive films can detect clinically relevant levels of heparin (0.5-5 units/mL) in undiluted human plasma. PMID- 9337593 TI - Study of 4-quinolone antibiotics in biological samples by short-column liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - Simultaneous detection and confirmation of 15 quinolone antibiotics was accomplished by fast short-column liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC/MS/MS). Several physiochemical parameters such as hydrophobicity and aqueous dissociation constants were calculated from the structural formulas of the quinolone drugs, and their impact on both chromatographic and mass spectrometric behavior was studied. Additionally, a possible influence of bulk solution pH on electrospray detection sensitivity of 4 quinolones was investigated and compared to predictions based on solution-phase equilibria. A signal intensity comparison of the MH+ ions at different pH values for all 15 compounds did not reveal any pH effect, despite variations by several orders of magnitude in equilibrium concentrations in bulk solution. To demonstrate the potential of the LC/MS/MS method, its application to trace analysis in several biological matrices such as milk, salmon, and human urine was investigated. The method was shown to be sensitive with detection limits down to 1 ppb in both milk and salmon tissue. The versatility of the method was also exhibited by utilizing it for rapid identification of urinary metabolites of ciprofloxacin. Finally, a new complementary approach is described for confirmatory analyses of 4-quinolones by means of a quasi-MS/MS/MS technique involving in-source collision-induced dissociation. It is shown that LC/quasi MS/MS/MS can significantly enhance structural information and, thus, the specificity of analysis for the investigated 4-quinolones. PMID- 9337594 TI - Formulation development and primary degradation pathways for recombinant human nerve growth factor. AB - The chemical and physical stabilities of recombinant human nerve growth factor (NGF) in aqueous solution were investigated between 5 and 37 degrees C and at pH 4.2-5.8. NGF chemical stability decreased with a decrease in pH due to Asp60 Pro61 cleavage, with the stability being greater in acetate buffer than in succinate buffer at each pH investigated. Aggregation was a significant degradation pathway at 37 degrees C, with the aggregation rate being greatest in succinate buffer at pH 5.8. Quantitation of NGF degradation by cation-exchange chromatography was complicated by the rearrangement of the NGF monomer variants into various mixed dimers over time. Treatment with dilute acid brought the dimer distribution rapidly to equilibrium, allowing NGF degradation to be accurately quantitated. An acetate-buffered formulation at pH 5.5 was investigated in more detail. To assist in degradation product identification, NGF degradation was accelerated with base, hydrogen peroxide, and temperature. These degradation products were shown to coelute on RP-HPLC with the variants found when the protein was stored at -70, 5, and 25 degrees C. By electrospray mass spectrometry, peptide maps, and LC/MS, these degradation products were shown to be monooxidized (Met37) and dioxidized (Met37 and Met92) NGF, with Met37 being more labile, deamidated NGF (Asn45), and NGF with Asp93 isomerized to beta-Asp93. NGF can be stored in pH 5.5 acetate buffer at 5 degrees C for 1.5 years with less than 10% conversion to these degradation products, with Asp93 isomerization being the primary degradation pathway. PMID- 9337595 TI - Discrimination of single-nucleotide polymorphisms in human DNA using peptide nucleic acid probes detected by MALDI-TOF mass spectrometry. AB - Human genomic and mitochondrial DNA contain large numbers of single-nucleotide polymorphisms (SNPs), many of which are linked to known diseases. Rapid and accurate genetic screening for important SNPs requires a general methodology which is easily implemented. We present here an approach to SNP discrimination based on high-specificity hybridization of peptide nucleic acid (PNA) probes to PCR-amplified DNA. The assay is directly applied to polymorphisms located within hypervariable region 1 of the human mitochondrial genome and type 1 suballeles of the human leukocyte antigen DQ alpha gene. Captured, single-stranded DNA molecules prepared by PCR amplification are hybridized with PNA probes in an allele-specific fashion. Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOFMS) is then used for rapid, precise, and unambiguous detection and identification of the hybridized PNA probes. Since PNA oligomers bind strongly to complementary DNA under minimal salt conditions, the use of PNA probes is compatible with MALDI-TOFMS. The unparalleled ability of MALDI-TOFMS analysis in terms of molecular weight resolution and accuracy, in conjunction with the highly specific PNA hybridization afforded by this method, offers promise for development into a multiplexed, high-throughput screening technique. PMID- 9337596 TI - An ion chromatographic method for measuring < 5 micrograms/L bromate ion in drinking water. AB - Many laboratories using the best available technology (ion chromatography) can only reliably measure BrO3- at the 10-20 micrograms/L level. Typically, BrO3- peak resolution is not a problem; however, finished water can contain appreciable amounts of Cl- that can interfere with BrO3- measurement using conductivity detection. Chlorpromazine is a selective reagent that does not react with Cl- and is an ideal postcolumn detection chemistry following IC separation. The chlorpromazine postcolumn method is linear over the 0.5-100 micrograms/L range. PMID- 9337597 TI - Hyphenation of ion exchange high-performance liquid chromatography with Fourier transform infrared detection for the determination of sugars in nonalcoholic beverages. AB - Fourier transform infrared spectroscopy (FT-IR) is presented here as a molecular specific detection system for high-performance liquid chromatography (HPLC) in an aqueous phase, focusing on the chromatographic separation of sugars in beverages. The separation was achieved with an isocratic HPLC setup using an ion exchange column (counterion, Ca2+). The FT-IR detection of the C-O bands in the mid-IR between 1000 and 1200 cm-1 was performed in real time with a 25 microns flow cell without elimination of the solvent. Characteristic FT-IR spectra of the common sugars sucrose, glucose, and fructose in concentrations of 1 mg/mL could be recorded during the separation. The calibration of these compounds in the 5-100 mg/mL range resulted in a linear correlation with a standard deviation of the method (Sx0) of 0.11, 0.07, and 0.11 mg/mL for sucrose, glucose, and fructose, respectively. The method was, furthermore, applied to the analysis of nine soft drinks and fruit juices containing between 6 and 97 mg/mL of each carbohydrate. The accuracy of the method was confirmed by standard ion exchange HPLC with refractive index detection. The average deviation from the reference method was in the range of 0.5-0.9 mg/mL. Furthermore, the method was found to be suitable to identify and quantify also minor components in beverages, such as taurine (4 mg/mL) and ethanol (0.4 mg/mL). PMID- 9337598 TI - Self-directed behavioral family intervention for parents of oppositional children in rural and remote areas. AB - Twenty-four parents of oppositional preschoolers were randomly assigned to either a self-directed behavioral family intervention condition (SD) or to a waitlist control group (WL). The self-directed parent training program, based on self regulation principles, consisted of a written information package and weekly telephone consultations for 10 weeks. At posttest, in comparison to the WL group, children in the SD group had lower levels of behavior problems on parent report measures of child behavior. At posttreatment, parents in the SD condition reported increased levels of parenting competence and lower levels of dysfunctional parenting practices as compared to parents in the WL condition. In addition, mothers reported lower levels of anxiety, depression, and stress as compared to mothers in the WL condition at posttreatment. Using mother's reports, gains in child behavior and parenting practices achieved at posttreatment were maintained at 4-month follow-up. PMID- 9337599 TI - The role of parental and extrafamilial social support in the psychosocial adjustment of children with a chronically ill father. AB - The relationships among illness stress, perceived support, and child psychosocial adjustment were examined for children living with a chronically ill father. Participants included fathers, mothers, and one child from 53 families in which the father had hemophilia and, in some cases, was HIV seropositive. Objective indicators of severity of illness and subjective measures of the physical and psychological impact of illness were used as sources of children's stress. Results indicated that the impact of illness, but not the severity of illness itself, related to child psychosocial adjustment. Main effects were observed for parental support on child- and parent-reported internalizing problems and stress buffering effects were obtained for parental support and extrafamilial support on parent-reported internalizing problems. Parental support also demonstrated a stress-buffering effect for child-reported depression. Assessment and intervention implications for behavioral clinicians and researchers are discussed. PMID- 9337600 TI - Psychological treatment of chronic posttraumatic stress disorder in victims of sexual aggression. AB - The aim of this research was to test the comparative effectiveness of two therapeutic modalities in the treatment of chronic posttraumatic stress disorder in victims of sexual aggression: (a) self-exposure and cognitive restructuring and (b) progressive relaxation training. The sample consisted of 20 patients (victims of rape in adulthood or adult victims of childhood sexual abuse) selected according to DSM-III-R criteria. A multigroup experimental design with repeated measures (pretreatment, posttreatment, and 1-, 3-, 6-, and 12-month follow-up) was used. Most treated patients improved, but the success rate was higher in all measures in the exposure and cognitive restructuring group immediately on posttreatment and at follow-up. Implications of this study for clinical practice and future research in this field are commented on. PMID- 9337601 TI - Adults' perceptions of the behavior of competent and dysfunctional children based on the children's physical appearance. AB - This study explored adults' judgments of competent and dysfunctional children's behavioral adjustment based solely on children's physical appearance. Adults rated photographs of preschoolers (6 boys, 6 girls) who previously had been classified as socially competent or dysfunctional on the basis of independent, standardized teacher ratings. Participants, who were not given any information about the stimulus children, rated their photographs one at a time on measures of attractiveness, aggression, anxiety, social competence, and overall adjustment. Results indicated that dysfunctional children were easily distinguished from their competent peers. Specifically, dysfunctional children were rated as less attractive, more aggressive, more anxious, less socially competent, and more likely to have an emotional or behavioral problem than competent children. These findings (especially strong for aggressive boys), remained significant when group differences in attractiveness were statistically controlled. Implications for interpreting the current literature on attractiveness and for modification of childhood behavior disorders are discussed. PMID- 9337602 TI - Decreasing disruptive behavior by adolescent boys in residential care by increasing their positive to negative interactional ratios. AB - An intervention for disruptive boys in residential care involving increases in positive to negative interactional ratios is described. The target of the intervention was daily problem behavior. Results from a pooled time series analysis of the data revealed a significant decrease in behavior problems (one problem per boy per day) during the intervention for the boys as a group. Results from comparisons of mean behavior problems during baseline and intervention revealed decreases for five of the six boys. Results from a multiple baseline across boys revealed experimental control for three of the six. The results are discussed in terms of response contingent reinforcement and systemic behavior analyses. The benefits of combined group and single subject data analyses are also discussed. PMID- 9337603 TI - Cognitive restructuring in the treatment of social phobia. Efficacy and mode of action. AB - Cognitive restructuring (CR) is commonly used to treat social phobia, although its contribution to treatment efficacy has not been established. CR requires the person to think about and discuss feared social events with his or her therapist and thus entails some degree of exposure to social stimuli. CR also is thought to enhance the efficacy of therapeutic exposure exercises (EXP). Four predictions were tested based on this model: Relative to a control intervention matched for the exposure inherent in CR, CR is more effective in (1) reducing social phobia, (2) reducing negative social cognitions, (3) increasing positive cognitions, and (4) enhancing the effects of subsequent EXP. People with generalized social phobia (N = 60) were randomly assigned to CR followed by EXP or to a control intervention followed by EXP. Support was found for predictions 1 to 3, but not 4. PMID- 9337604 TI - Residential behavior therapy for children with conduct disorders. AB - Children with conduct disorders are often referred to residential treatment centers (RTCs). RTCs shorten the length of treatment and thus feel they need to reconceptualize the purpose and process of treatment. Two intervention strategies have been found to affect conduct disorder in outpatient settings: parent training programs that are based on operant learning principles and cognitive behavioral programs that focus on the relation between cognition and behavior. These strategies should not be transferred to the RTC but adapted to the characteristics of residentially treated conduct disordered children and their parents. These methods should be used together to integrate and strengthen the various learning processes that residential treatment can foster. An outline is given of a comprehensive and integrated residential treatment program based on behavioral methods that have been proven to affect conduct disorder. PMID- 9337605 TI - Evaluating behavioral techniques in training individuals with severe and profound mental retardation to use functional independent living skills. AB - Three treatment approaches were evaluated for functional skill acquisition in individuals with severe and profound mental retardation. The control condition comprised the standard treatment protocol: verbal prompting, modeling, and physical guidance. The first condition added the components of staff training, feedback, and edible reinforcement for clients. The second treatment condition supplemented the first by the addition of verbal and edible reinforcement for staff. Subjects included 30 residents from a large developmental center (Pinecrest) in central Louisiana. The control protocol proved to be an ineffective training regimen. Experimental 1 led to statistically significant increase in learning when compared to controls. Experimental 2 led to additional statistically significant improvements beyond those achieved by Experimental 1. Daily documentation was once again shown not to enhance treatment effectiveness. Implications of the findings are discussed. PMID- 9337606 TI - Effects of training in procedural justice on perceptions of disciplinary fairness by unionized employees and disciplinary subject matter experts. AB - A training program, based on procedural justice theory, was developed for teaching supervisors to take effective disciplinary action with employees. Canadian supervisors of unionized employees were randomly assigned to the training (n = 35) or the control group (n = 36). Analyses of variance revealed that both supervisory self-efficacy and outcome expectancies were significantly higher in the training than in the control conditions. Following simulated role play exercises derived from organizational incidents, both unionized employees and disciplinary subject matter experts (managers, union officials, and attorneys) rated the trained supervisors higher on disciplinary fairness behavior than the supervisors in the control group. Self-efficacy was found to mediate the relationship between training and perceptions of disciplinary fairness. PMID- 9337607 TI - Antecedents and outcomes of coping behaviors among unemployed and reemployed individuals. AB - This study examined 3 predictors (self-esteem, perceived control, and optimism) and 3 outcomes (short-term mental health, reemployment, and long-term mental health) of coping behavior among unemployed individuals in a longitudinal context. The predictors and outcomes had differential relationships with the 5 coping behaviors (proactive search, nonwork organization, positive self assessment, distancing from loss, and job devaluation) assessed. Two interactions between coping behavior and perceived situational control (the perception that one would find a job) were found. Proactive search (job-search behavior) was associated with decreased mental health at Time 1 among individuals with low situational control but not among individuals with high situational control. Distancing from loss was associated with decreased reemployment among individuals with low situational control but not among individuals with high situational control. PMID- 9337608 TI - Relationship between conscientiousness and learning in employee training: mediating influences of self-deception and self-efficacy. AB - A field study of 97 employees tested a model of the mediating influences of self deception and task-specific self-efficacy in the relationship between conscientiousness and learning. The setting was an introductory Windows 3.1 software training course. Findings indicated that, as hypothesized, self deception and self-efficacy mediated the relationship between conscientiousness and learning. Specifically, conscientiousness was positively related to self deception, which was negatively related to learning, and conscientiousness was positively related to self-efficacy, which was positively related to learning. In addition, 4 alternative models were estimated. The results of the tests of the 4 alternative models were not supported by the data, further substantiating the validity of the hypothesized model. PMID- 9337609 TI - Excess coffee consumption in simulated complex work settings: detriment or facilitation of performance? AB - Twenty-four managers who normally consume between 400 and 1,000 mg of caffeine per day participated in all-day quasi-experimental simulations. In a crossover, doubleblind design, they made complex managerial decisions either on treatment with their typical daily dose of caffeine or on treatment with 400 mg of caffeine in excess of daily consumption. The effect of caffeine treatment on various validated performance indicators was investigated. The impact of excess caffeine consumption was mild. Increased caffeine facilitated speed of response to incoming information but decreased utilization of opportunity. No significance was obtained for other measures of managerial effectiveness (such as activity, breadth, strategy, and emergency response). PMID- 9337610 TI - Perceived organizational support, discretionary treatment, and job satisfaction. AB - A diverse sample of 295 employees drawn from a variety of organizations was surveyed to investigate (a) whether the relationship between the favorableness of job conditions and perceived organizational support (POS) depends on employee perceptions concerning the organization's freedom of action and (b) whether POS and overall job satisfaction are distinct constructs. The favorableness of high discretion job conditions was found to be much more closely associated with POS than was the favorableness of low-discretion job conditions. No such relationship was found between job conditions and satisfaction. To decide how much the organization values their contributions and well-being, employees distinguish job conditions whose favorableness the organization readily controls versus job conditions whose favorableness is constrained by limits on the organization's discretion. PMID- 9337611 TI - Meso-tetraphenylporphyrin dimer derivative as a potential photosensitizer in photodynamic therapy. AB - Studies on the synthesis, preliminary in vivo biological activity, singlet oxygen and fluorescence yields of a dimeric porphyrin (D1) are described. The pharmacokinetic behavior and photodynamic properties of the dimer D1 were examined in Balb/c mice bearing an MS-2 fibrosarcoma. Compound D1 shows a high selectivity for tumor localization (tumor/peritumoral tissue ratios of dye concentration ranging between ca 100 and 140 at 24 h after drug administration of 5.0-1.0 mg kg-1 into DL-alpha-dipalmitoylphosphatidylcholine liposomes). The phototherapeutic efficiency of dimer D1 was evaluated by following the growth curves of fibrosarcoma irradiated with red light (600-700 nm) with a total dose of 400 J cm-2, at 24 h after intravenous injection. Photodynamic therapy-treated tumors showed a significant delay in growth as compared to untreated control mice. The results obtained suggest that the porphyrin dimer D1 may be a promising candidate for further use in PDT experiments. PMID- 9337612 TI - Distinctive molecular abnormalities in benign and malignant skin lesions: studies by Raman spectroscopy. AB - Near-infrared Fourier transform Raman spectroscopy is an analytical, nondestructive technique that provides information about the molecular structure of the investigated sample. The molecular structure of proteins and lipids differ between neoplastic and normal tissues and therefore Raman spectroscopy has been considered promising for the diagnosis of cancer. We aimed to compare the molecular structure of normal skin, benign and malignant skin lesions by the near infrared Fourier transform Raman spectroscopy. Biopsies were obtained from the following skin lesions: skin tag, dermatofibroma, seborrhoeic keratosis, actinic keratosis, keratoacanthoma, basal cell carcinoma, squamous cell carcinoma, nevus intradermalis, nevus compositus, dysplastic nevus and lentigo maligna. Control skin was harvested from the vicinity of these lesions. In the Raman spectra, the secondary structure of the proteins was reflected by the amide vibrations of peptide bonds. The principal lipid vibrations were twisting and wagging (CH2) and CH stretching vibrations. Histologically distinguishable lesions showed specific combinations of band changes indicating alterations in the protein conformation and in the molecular structure of the lipids. Histogenetically related lesions (actinic keratosis and sqamous cell carcinoma) produced similar but not identical patterns of spectral changes. Because the examined skin lesions produced reproducible and unique spectra, we suggest that Raman spectroscopy will be useful for diagnosis of skin lesions. PMID- 9337613 TI - Photodegradation and photobinding of tiaprofenic acid: in vitro versus in vivo. AB - The photoreactivity of the photosensitizing nonsteroidal anti-inflammatory drug tiaprofenic acid (TA) and its photoproduct decarboxytiaprofenic acid (DTA) was studied both in the presence and in the absence of bovine serum albumin (BSA). The photoproduct DTA was found to be photostable in buffered aqueous solution at pH 7.4, but photodecomposed when BSA was present in the reaction medium. Both TA and DTA underwent irreversible photobinding to BSA in an almost quantitative way, as evidenced by radioactivity measurements using labeled (3H) compounds. In the case of TA, it has been proven that photobinding is mainly attributable to the phototoreactivity of in situ-generated DTA. The photodegradation and photobinding of TA were also investigated in the epidermis in vivo. Rats were exposed to UVA after application of TA to their shaven dorsal skin. During the initial periods of irradiation, the amount of TA decreased sharply, and the yield of the corresponding photoproduct (DTA) reached a maximum. Prolonged irradiation led to photodegradation of DTA. In vivo photobinding was studied using 3H-TA. Photobinding took place slowly at the beginning, but its rate increased sharply after complete photoconversion of TA, when the photoproduct DTA reached the maximum concentration. Thereafter, the decrease of DTA was more pronounced than that of TA. This indicates that-also in vivo-DTA rather than TA is responsible for the photobinding to biomacromolecules in the viable layer of the epidermis. Overall, the above results suggest that irradiation of TA in buffered aqueous solution, in the presence of proteins, is a reasonable model system to study the photodegradation and photobinding behavior of this drug in vivo. From the qualitative point of view, the main conclusion is that DTA plays a key role both in vivo and in vitro: it is the major photoproduct, it undergoes further photodegradation upon prolonged irradiation, and it appears to be responsible for the photobinding process. PMID- 9337614 TI - Photochemistry of water-soluble quinones. Production of a water-derived spin adduct. AB - The photolyses of phosphate-buffered (pH 7) air- and nitrogen-saturated solutions containing the water-soluble quinones, 1,4-benzoquinone (BQ), 2-methyl-1,4 benzoquinone (MBQ), sodium 1,4-naphthoquinone-2-sulfonate (NQ2S), 9,10 anthraquinone-2-sulfonate (AQ2S) or 9,10-anthraquinone-1,5-disulfonate (AQDS), and the spin trap 5,5-dimethylpyrroline-1-oxide (DMPO) produce a DMPO-OH adduct. Electron paramagnetic resonance spectroscopy of the photolyzed samples in 17O enriched water demonstrates that this adduct derives almost exclusively from water. With the exception of BQ, quantum yields for the formation of DMPO-OH are larger in air than in nitrogen-saturated samples, thus supporting the idea of the formation of air-oxidized intermediates that enhance the DMPO hydroxylation reaction rate. Evidence has been obtained which suggests that BQ and MBQ, but not AQDS, are able to photooxidize water, with the consequent production of the free OH radical. PMID- 9337615 TI - Light emission resulting from hydroxylamine-induced singlet oxygen formation of oxidizing LDL particles. AB - Oxidation of low-density lipoprotein (LDL) by low amounts of cupric ions resulted in the formation of singlet oxygen (1O2, 1 delta g) when hydroxylamine (NH2OH) was added. Direct evidence on this excited species came from partial spectral resolution of the emitted light in the red spectral region (634 nm and 703 nm), which can be attributed to the dimol decay of singlet oxygen. Additional evidence for the existence of singlet oxygen came from the enhancing effect of deuterium oxide buffer (D2O) on chemiluminescence intensity and the quenching effect of sodium azide. A linear correlation between NH2OH-dependent chemiluminescence intensity and the amount of diene conjugates (DC) formed in this reaction was observed. Removal of adventitious transition metals by adequate chelators prevented chemiluminescence in this system; NH2OH was also found to efficiently decrease metabolites of lipid peroxidation (LPO). Our findings are consistent with a sequence of reactions in which NH2OH first converts transition metals to their reduced state, thereby stimulating the formation of alkoxy- and peroxyradicals. Peroxyradicals decompose in a bimolecular Russel reaction to hydroxyl compounds and singlet oxygen while the majority of alkoxy radicals are eliminated by a secondary reaction with NH2OH. Identical effects were observed when reducing antioxidants such as ascorbic acid or trolox C were used instead of hydroxylamine. PMID- 9337616 TI - Development of refractoriness of induced human heme oxygenase-1 gene expression to reinduction by UVA irradiation and hemin. AB - Using primary human fibroblasts we have observed the existence of an acquired refractoriness of the heme oxygenase-1 gene to induction by a second dose of UVA (320-380 nm) radiation. We studied the kinetics of development of refractoriness over a time interval of up to 72 h between the first inducing event and the second (challenge) dose. Complete refractoriness was observed at 48 h. We also studied development of refractoriness after UVA, sodium arsenite and H2O2 treatment in all possible combinations and demonstrated that only UVA led to refractoriness. Ultraviolet radiation induced partial refractoriness to H2O2 induction but did not change the response to sodium arsenite. In an investigation of the mechanism of development of refractoriness we used the heme oxygenase inhibitor, tin-protoporphyrin IX and showed that induction of heme oxygenase enzymatic activity is a crucial step. However, the induction of ferritin, which is known to play a key role in protection against oxidative stress, did not appear to be involved. Damage to membranes is also probably not involved in the refractoriness mechanism. Because either hemin alone or UVA radiation are able to lead to a refractoriness of the heme oxygenase-1 gene to reinduction by a second exposure to one or the other agent in human fibroblasts, we conclude that heme, or an as yet unidentified heme derivative, is involved in the refractoriness response. PMID- 9337617 TI - Action spectrum for induction of promoter activity of phenylalanine ammonia-lyase gene by UV in carrot suspension cells. AB - The full-length promoter (-2335) of the carrot (Daucus carota) phenylalanine ammonia-lyase gene (gDcPAL1) fused to the luciferase reporter gene was transiently transformed to carrot protoplasts by electroporation, and the promoter activity induced by monochromatic UV light of various wavelengths was examined. The action spectrum constructed from the fluence-response curves showed a single peak at around 280 nm, suggesting that the activation of the gDcPAL1 promoter is categorizable as one of the UVB light responses. The same assay system was applied to variously truncated gDcPAL1 promoters and to CaMV35S promoter fusion with various parts 5'-upstream of the gDcPAL1 promoter. The region from -396 to -190 (relative to the transcription start site) fused to the CaMV35S core (-90) promoter showed a 280 nm-dominant response. However, gDcPAL1 promoters truncated above -570 and -396, although they contain the region between -396 and -190, did not show such a typical UVB response, i.e. they responded to 260 nm light as much as to 280 nm light. The promoter truncated to below -190 also responded to 260 nm light as much as to 280 nm light. Therefore we assumed that the gDcPAL1 promoter is composed of three functionally different parts: the upstream above -570 (modulator), the region from -396 to -190 (UVB responsive) and the down-stream below -190 (UVB and C responsive). The overall UVB response of the gDcPAL1 full-length promoter is explained as the result of interaction of these three components. PMID- 9337618 TI - Initiation of apoptosis versus necrosis by photodynamic therapy with chloroaluminum phthalocyanine. AB - While chloroaluminum phthalocyanine is a highly effective photosensitizer of murine leukemia P388 or L1210 cells, the mode of cell death varies as a function of the PDT dose. When cells were incubated with 0.3 microM of the sensitizer, a light dose of 45 mJ cm-2 (670 +/- 5 nm) yielded a 90% apoptotic cell population within 60 min. The sensitizer localized throughout the cytoplasm and catalyzed both lysosomal and mitochondrial photodamage at this light dose. Higher light doses yielded progressively more membrane photodamage and inhibited the apoptotic response as determined by the examination of Hochst dye HO 33342-labeled nuclei, DNA fragmentation on gels and a poly(adenosylribose) polymerase (PARP)-cleavage assay. Pulse-field gel electrophoresis revealed nonspecific DNA degradation to particles > or = 50 kbp at the higher PDT doses but neither PARP cleavage nor apoptotic nuclei. PMID- 9337619 TI - The effects of UVA-I (340-400 nm), UVA-II (320-340 nm) and UVA-I+II on the photoisomerization of urocanic acid in vivo. AB - Ultraviolet B radiation (280-320 nm) can systemically suppress contact hypersensitivity (CHS), delayed type hypersensitivity (DTH) and tumor rejection responses in mice. Several models have been postulated for the initiation of this UVB-induced immune suppression and, although the complete mechanism is unclear, our early studies suggested that initiation is via the activation of a photoreceptor in the skin, identified as urocanic acid (UCA). Recent preliminary data from our laboratory and others indicated that UVA (320-400 nm)-emitting broad-band sunlamps can also isomerize UCA but may not lead to immune suppression, in contrast to UVB-emitting sunlamps, which cause both effects. Although the reason for this inconsistency is unknown, the emission spectra of UVA lamps contain differing amounts of UVB, UVA-I (340-400 nm) and UVA-II (320 340 nm) from those of UVB sources. In this study we determined a detailed dose response for the isomerization of UCA in mouse skin using the UVA-I, UVA-II and UVA-I+II wavelength ranges. The dose-response curves obtained were put on an equal energy basis by quantum correction and the possibility of wavelength interaction for this effect investigated. A simple additive wavelength interaction between UVA-I, UVA-II, and UVA-I+II was observed for trans-UCA photoisomerization. This result indicates that the failure of UVA-I, UVA-II or UVA-I+II radiation to induce immune suppression of the CHS response in an animal model is not due to complex wavelength interactions and/or the presence of an in vivo endogenous photosensitizer of UCA isomerization. Other factors, such as downstream blocking by UVA of the cis-UCA generated signal, may be involved. PMID- 9337620 TI - Photodynamic therapy with topical delta-aminolevulinic acid delays UV photocarcinogenesis in hairless mice. AB - Photodynamic therapy (PDT) with topical application of delta-aminolevulinic acid (ALA) followed by irradiation with visible light (ALA-PDT) is a relatively new and promising experimental treatment of superficial premalignant and malignant skin neoplasms. The purpose of this study was to determine whether ALA-PDT can prevent photocarcinogenesis in hairless mice exposed to solar UV. A total of 140 mice was divided into seven groups of 20 mice each. Group 1: solar-UV exposure. Group 2: solar UV and a cream base+visible light once a week. Group 3: solar UV and ALA-PDT once a week. Group 4: solar UV and ALA-PDT once every second week. Group 5: solar UV and ALA-PDT every fourth week. Group 6: ALA-PDT once a week. Group 7: no treatment. The time to first and to second tumor > or = 1 mm was registered. Predefined endpoints, such as one tumor > or = 4 mm or an area of small confluent tumors on the back of the mice were criteria for withdrawal from the experiment. The time to first and to second tumor was significantly longer in the ALA-PDT-treated mice than in mice only exposed to solar UV and solar-UV/cream base-visible light (P < 0.005). However, we observed an increased death and accident rate in the ALA-PDT-treated groups compared to the groups not treated with ALA-PDT (chi-square test, P = 0.0250). Significantly more ALA-PDT-treated mice were withdrawn because of a tumor > or = 4 mm (P = 0.0005). The UV unexposed mice developed no tumors. Repetitive treatments with ALA-PDT delay photoinduced carcinogenesis in mice. PMID- 9337621 TI - Oxyradical-mediated clastogenic plasma factors in psoriasis: increase in clastogenic activity after PUVA. AB - Psoriasis is a common skin disorder characterized by hyperproliferation and incomplete differentiation of epidermal keratinocytes. Psoralen plus UVA (PUVA) is one of the treatments proposed for this disease. We had reported previously that exposure of regular blood cultures from healthy donors to PUVA leads to chromosomal breakage via the formation of transferable clastogenic materials, a phenomenon inhibitable by superoxide dismutase. In the present paper we show that these clastogenic factors (CF) are also formed in vivo. The CF were found in about 50% of the psoriasis patients studied (14 out of 31). In PUVA-treated psoriasis patients, the clastogenic activity of the plasma increased significantly between the first and the last (16th) exposure to PUVA. We hypothesize that CF formation in psoriasis is similar to that in other diseases accompanied by oxidative stress, in particular chronic inflammatory diseases with autoimmune reactions such as lupus erythematosus, progressive systemic sclerosis, rheumatoid arthritis and others. Increased superoxide production by phagocytes, formation of lipid peroxidation products and release of cytokines are considered to be responsible for the superoxide-stimulating and chromosome-damaging properties of patients' plasma. During PUVA therapy, superoxide generated via the interaction of psoralen with UVA may contribute to CF formation in addition to superoxide from inflammatory cells. An increased risk of cancer and leukemia is observed in diseases accompanied by CF formation. Therefore CF may contribute to the well-known risk of photocarcinogenesis by PUVA therapy. This additional risk may be preventable by antioxidants and superoxide scavengers. PMID- 9337622 TI - Electron spin resonance evidence of the generation of superoxide anion, hydroxyl radical and singlet oxygen during the photohemolysis of human erythrocytes with bacteriochlorin a. AB - Photodynamic therapy with bacteriochlorin a (BCA) as sensitizer induces damage to red blood cells in vivo. To assess the extent of the contributuion of reactive oxygen species (ROS) and to determine a possible reaction mechanism, competition experiments with assorted ROS quenching or/and enhancing agents were performed in human erythrocytes as model system and in phosphate buffer. In the erythrocyte experiments, a 2% suspension was incubated with BCA for 1 h, washed with phosphate-buffered saline, resuspended and subsequently illuminated with a diode laser using a fluence rate of 2.65 mW/cm2. Potassium leakage and hemolysis were light and BCA dose dependent. Adding tryptophan (3.3 mM), azide (1 mM) or histidine (10 mM) to the erythrocyte suspension before illumination delayed the onset of K-leakage and hemolysis suggesting a type II mechanism. The D2O did not affect K-leakage nor photohemolysis. Adding mannitol (13.3 mM) or glycerol (300 nM) also caused a delay in the onset of K-leakage and hemolysis, suggesting the involvement of radicals. In phosphate buffer experiments, it was shown using electron spin resonance (ESR) associated with spin-trapping techniques that BCA is able to generate O2-. and OH. radicals without production of aqueous electron. Visible or UV irradiation of the dye in the presence of the spin trap 5,5 dimethyl-1-pyrroline-N-oxide (DMPO) gave an ESR spectrum characteristic of the DMPO-hydroxyl radical spin adduct DMPO-OH. Addition of ethanol or sodium formate produced supplementary hyperfine splittings due to the respective CH3CHOH. and CO2-. radical adducts, indicating the presence of free OH.. Production of DMPO-OH was partly inhibited by superoxide dismutase (SOD), catalase and desferrioxamine, suggesting that the iron-catalyzed decomposition of H2O2 was partly involved in the formation of one part of the observed OH.. The complementary inhibition of DMPO-OH production by azide and 9,10-anthracenedipropionic acid (ADPA) was consistent with 1O2 production by BCA followed by reaction of 1O2 with DMPO and decay of the intermediate complex to form DMPO-OH and free OH.. All our results seem to indicate that BCA is a 50%/50% type 1/type 2 sensitizer in buffered aqueous solutions and confirmed that the dye-induced hemolysis of erythrocytes was cell caused by a mixed type 1/type 2 mechanism. PMID- 9337623 TI - Photodynamic treatment of red blood cell concentrates for virus inactivation enhances red blood cell aggregation: protection with antioxidants. AB - Photodynamic treatment (PDT) using phthalocyanines and red light appears to be a promising procedure for decontamination of red blood cell (RBC) concentrates for transfusion. A possible complication of this treatment may be induced aggregation of RBC. The production of RBC aggregates was measured with a novel computerized cell flow properties analyzer (CFA). The PDT of RBC concentrates with sulfonated aluminum phthalocyanine (AIPcS4) and the silicon phthalocyanine Pc 4 under virucidal conditions markedly enhanced RBC aggregation and higher shear stress was required to disperse these aggregates. The clusters of cells were huge and abnormally shaped, unlike the rouleaux formed by untreated RBC. This aggregation was prevented when a mixture of antioxidants was included during PDT. Addition of the antioxidants after PDT reduced aggregation only partially. It is concluded that inclusion of antioxidants during PDT of RBC concentrates prior to transfusion may reduce or eliminate the hemodynamic risk that the virucidal treatment may present to the recipient. PMID- 9337624 TI - The effects of thrombocytopenia on vessel stasis and macromolecular leakage after photodynamic therapy using photofrin. AB - Several studies have reported thrombus formation and/or the release of specific vasoactive eicosanoids, suggesting that platelet activation or damage after photodynamic therapy (PDT) may contribute to blood flow stasis. The role of circulating platelets on blood flow stasis and vascular leakage of macromolecules during and after PDT was assessed in an intravital animal model. Sprague-Dawley rats bearing chondrosarcoma on the right hind limb were injected intravenously (i.v.) with 25 mg/kg Photofrin 24 h before light treatment of 135 J/cm2 at 630 nm. Thrombocytopenia was induced in animals by administration of 3.75 mg/kg of rabbit anti-rat platelet antibody i.v. 30 min before the initiation of the light treatment. This regimen reduced circulating platelet levels from 300,000/mm3 to 20,000/mm3. Reductions in the luminal diameter of the microvasculature in normal muscle and tumor were observed in control animals given Photofrin and light. Venule leakage of macromolecules was noted shortly after the start of light treatment and continued throughout the period of observation. Animals made thrombocytopenic showed none of these changes after PDT in either normal tissues or tumor. The lack of vessel response correlated with the absence of thromboxane release in blood during PDT. These data suggest that platelets and eicosanoid release are necessary for vessel constriction and blood flow stasis after PDT using Photofrin. PMID- 9337625 TI - Fundamental differences of excitation spectrum between malignant and benign breast tissues. AB - Differences observed in the UV excitation spectrum of malignant and benign breast tissues are used to distinguish between malignant and benign breast tissues. These changes are attributed to changes with specialized proteins. PMID- 9337626 TI - Visualizing gene expression in living mammals using a bioluminescent reporter. AB - Control of gene expression often involves an interwoven set of regulatory processes. As information regarding regulatory pathways may be lost in ex vivo analyses, we used bioluminescence to monitor gene expression in living mammals. Viral promoters fused to firefly luciferase as transgenes in mice allowed external monitoring of gene expression both superficially and in deep tissues. In vivo bioluminescence was detectable using either intensified or cooled charge coupled device cameras, and could be detected following both topical and systemic delivery of substrate. In vivo control of the promoter from the human immunodeficiency virus was demonstrated. As a model for DNA-based therapies and vaccines, in vivo transfection of a luciferase expression vector (SV-40 promoter and enhancer controlling expression) was detected. We conclude that gene regulation, DNA delivery and expression can now be noninvasively monitored in living mammals using a luciferase reporter. Thus, real-time, noninvasive study of gene expression in living animal models for human development and disease is possible. PMID- 9337627 TI - An insertion or deletion in the extramembrane loop connecting helices E and F of archaerhodopsin-1 affects in vitro refolding and slows the photocycle. AB - Upon addition of retinal, archaeopsin-1 expressed in Escherichia coli (ecaO-1002) regenerated the chromophore in dimyristoyl phosphatidylcholine (DMPC), 3-[(3 cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS) and sodium dodecyl sulfate (SDS) mixed micelles as efficiently as the same opsin prepared from halobacteria. Introduction of an insertion or a deletion of five amino acids into the surface loop connecting helices E and F changed the secondary and tertiary structures of ecaO-1002 in SDS, and diminished regeneration of the chromophore. The effect of the insertion and deletion on the in vitro refolding was specific to archaeopsin because the same insertion introduced at the corresponding position of bacterioopsin (bO) did not affect chromophore regeneration. The photocycle of the regenerated ecaR-1002 decreased in DMPC/CHAPS/SDS mixed micelles compared with that of aR-1 in the claret membrane, which was consistent with the reported behavior of bO. Unexpectedly, the insertion and deletion in loop EF perturbed the photocycle of the regenerated ecaR-1002. The accumulation of long-lived N- and O-like intermediates suggested that the insertion and deletion slowed down the proton uptake steps at the cytoplasmic surface. PMID- 9337628 TI - Schizophrenia: a neural diathesis-stress model. AB - There is a substantive literature on the behavioral effects of psychosocial stressors on schizophrenia. More recently, research has been conducted on neurohormonal indicators of stress responsivity, particularly cortisol release resulting from activation of the hypothalamic-pituitary-adrenal (HPA) axis. This article integrates the psychosocial and biological literatures on stress in schizophrenia, and it offers specific hypotheses about the neural mechanisms involved in the effects of stressors on the diathesis. Both the behavioral and biological data indicate that stress worsens symptoms and that the diathesis is associated with a heightened response to stressors. A neural mechanism for these phenomena is suggested by the augmenting effect of the HPA axis on dopamine (DA) synthesis and receptors. Assuming the diathesis for schizophrenia involves an abnormality in DA receptors, it is proposed that the HPA axis acts as a potentiating system by means of its effects on DA. At the same time, DA receptor abnormality and hippocampal damage render the patient hypersensitive to stress. This neural diathesis-stress model is consistent with findings on prenatal factors and brain abnormalities in schizophrenia, and it provides a framework for explaining some key features of the developmental course and clinical presentation. PMID- 9337629 TI - Rethinking infant knowledge: toward an adaptive process account of successes and failures in object permanence tasks. AB - Infants seem sensitive to hidden objects in habituation tasks at 3.5 months but fail to retrieve hidden objects until 8 months. The authors first consider principle-based accounts of these successes and failures, in which early successes imply knowledge of principles and failures are attributed to ancillary deficits. One account is that infants younger than 8 months have the object permanence principle but lack means-ends abilities. To test this, 7-month-olds were trained on means-ends behaviors and were tested on retrieval of visible and occluded toys. Means-ends demands were the same, yet infants made more toy-guided retrievals in the visible case. The authors offer an adaptive process account in which knowledge is graded and embedded in specific behavioral processes. Simulation models that learn gradually to represent occluded objects show how this approach can account for success and failure in object permanence tasks without assuming principles and ancillary deficits. PMID- 9337630 TI - The genetics and evolution of handedness. AB - At some point in hominid evolution, a mutation may have produced a "dextral" (D) allele, strongly biasing handedness in favor of the right hand and control of speech toward the left cerebral hemisphere. An alternative (chance [C]) allele is presumed directionally neutral, although there are probably other genes that influence asymmetries and that may create a weak bias toward right-handedness (and other asymmetries). Simulations show that the D allele could have spread quite quickly through a population, given even a minuscule advantage of CD heterozygotes over CC and DD homozygotes in terms of reproductive fitness. This heterozygotic advantage would also explain the apparent stability in the relative proportions of left-handers and right-handers. This putative, uniquely human allele may have emerged with the evolution of Homo sapiens in Africa some 150,000 to 200,000 years ago. PMID- 9337631 TI - Lexical access in aphasic and nonaphasic speakers. AB - An interactive 2-step theory of lexical retrieval was applied to the picture naming error patterns of aphasic and nonaphasic speakers. The theory uses spreading activation in a lexical network to accomplish the mapping between the conceptual representation of an object and the phonological form of the word naming the object. A model developed from the theory was parameterized to fit normal error patterns. It was then "lesioned" by globally altering its connection weight, decay rates, or both to provide fits to the error patterns of 21 fluent aphasic patients. These fits were then used to derive predictions about the influence of syntactic categories on patient errors, the effect of phonology on semantic errors, error patterns after recovery, and patient performance on a single-word repetition task. The predictions were confirmed. It is argued that simple quantitative alterations to a normal processing model can explain much of the variety among patient patterns in naming. PMID- 9337632 TI - A simple versatile method for measuring tail cuff systolic blood pressure in conscious rats. AB - 1. The non-invasive measurement of tail cuff systolic blood pressure in conscious rats is routinely used in long-term cardiovascular studies. There are a number of commercially available tail cuff systems, however, these apparatus are generally expensive and are dedicated for single-task operations. In the present study, a simple method for measuring systolic blood pressure, which requires only minor modifications to the existing hardware found in most cardiovascular laboratories, is described. 2. Systolic blood pressure measurements were made in the conventional manner by determining the systolic blood pressure which coincided with the restoration of the caudal artery pulse. This was achieved by using an inexpensive piezo-electric pulse transducer to detect the pulse, and this was coupled to a standard data-acquisition system (MacLab, ADInstruments) normally set up to record blood pressure. This method was compared with another established tail cuff method, as well as with direct intra-arterial recordings. 3. It was found that the results obtained using both tail cuff systems were in good agreement when systolic blood pressure was measured in Wistar-Kyoto rats and spontaneously hypertensive rats. In addition, systolic blood pressure was measured over 4 weeks in 2K1C rats and sham-operated rats, with both tail cuff methods producing similar results, which were not significantly different from direct intra-arterial recordings in the same animals. 4. Thus, in the present study, with only minor modifications, the same equipment was used for both direct and indirect determinations of systolic blood pressure. This situation differs from other conventional tail cuff systems since these items are designed for a single purpose. Therefore, the current method using piezo-electric sensor/MacLab technology should be viewed as a relatively simple, flexible and cheap alternative method to measure tail cuff systolic blood pressure in conscious rats. PMID- 9337633 TI - Non-invasive measurement of cardiac output and ventricular ejection fractions in chronic cardiac failure: relationship to impaired exercise tolerance. AB - 1. The role of cardiac output limitation in the pathophysiology of exercise in patients with chronic failure remains undefined. During steady-state submaximal exercise, oxygen uptake is similar in patients and control subjects, but it is not known if cardiac output is also similar. We wished to determine if the reduced exercise tolerance of patients with chronic cardiac failure during such exercise is related to reduced cardiac output, or to peripheral factors. 2. Ten male patients with stable chronic failure and ten age-matched male normal controls were studied at rest and during exercise. Each subject performed a familiarization exercise test, a symptom-limited maximal exercise test and two submaximal exercise tests. Cardiac output was measured by a carbon dioxide rebreathing method. We also measured oxygen consumption, ventilation, Borg score of perceived exertion and venous lactate concentration, and ejection fractions. 3. As expected, patients had lower peak oxygen consumption [median (range) 1.18 (0.98-1.76) versus 1.935 (1.53-2.31) l/min; P < 0.001], lower peak venous lactate concentration but a similar overall level of perceived exertion. At the same submaximal workload, patients and control subjects had similar oxygen consumption [0.67 (0.59-0.80) versus 0.62 (0.52-0.82) l/min] and cardiac output [6.92 (5.79 9.76) versus 7.3 (5.99-10.38) l/min] but the patients had a greater perceived level of exertion [Borg score: 4 (1-6) versus 3 (1-5); P < 0.005], higher venous lactate concentration [1.6 (1-3.3) versus 1.14 (0.7-1.7) mmol/l; P < 0.05] and higher heart rate [106 (89-135) versus 87 (69-112) beats/ min; P < 0.005]. 4. During submaximal exercise at a similar absolute workload, patients with cardiac failure have a similar oxygen uptake and cardiac output but greater anaerobiosis and increased fatigue when compared with normal subjects. These findings appear to relate predominantly to changes that occur in the periphery rather than abnormalities of central cardiac function. PMID- 9337634 TI - Variability of near-fainting responses in healthy 6-16-year-old subjects. AB - 1. Fainting is a common phenomenon in young subjects, but the final events before the actual faint are not well known. The aim of the present study was to study the inter-individual variability of haemodynamic events associated with near fainting in children and teenagers. 2. Sixty-eight healthy subjects (aged 6-16 years) performed a 70 degrees tilt-up test with intravascular instrumentation for 5 min. Responses in 29 near-fainting subjects were analysed and compared with 39 non-fainting subjects. Arterial pressure was measured by Finapres. Left ventricular stroke volume was computed from the pressure pulsation waveform. 3. Inability to maintain vasomotor tone was the mechanism underlying near-fainting in the vast majority of near-fainting subjects. The three classical haemodynamic responses (vasovagal, vasodepressor and vagal) could be recognized, but large individual differences were found. After tilt back, blood pressure in near fainters showed a mirror response to the stage before tilt-back; blood pressure gradually increased and was normal at 1 min after tilt-back. 4. The variability in haemodynamic responses on approach of an orthostatic faint is wide in the young. PMID- 9337635 TI - Beneficial effect of the calcium-sensitizing drug EMD 57033 in a canine model of dilated heart failure. AB - 1. Pacing-induced heart failure was studied in eight dogs. Heart failure was induced by right ventricular pacing at 250-260 beats/min for 6 weeks. Evidence of heart failure was determined clinically and by measurement of left ventricular (LV) dimensions by transoesophageal echocardiography. 2. Haemodynamic measurements of LV pressure, maximum rate of rise of LV pressure (LVdP/dtmax), cardiac output, mean arterial pressure, heart rate, pulmonary artery and pulmonary wedge pressures were made during infusion of solvent (control) and the calcium sensitizer EMD 57033 (0.6 mg min-1 kg-1). 3. The degree of heart failure varied from mild to severe in different individuals, but in each case EMD 57033 exerted a positive inotropic effect on LV haemodynamics and dimension. 4. The positive inotropic effect of the calcium sensitizer was manifest by increased peak LVdP/dt with a subsequent increase in cardiac output at the same mean arterial pressure. 5. This study clearly demonstrates that there is the potential for improvement of contractility of the failing myocardium of the intact mammal by an agent with a mechanism of action which does not involve an increase in intracellular calcium. PMID- 9337636 TI - Hyperdynamic circulation of cirrhotic rats with ascites: role of endotoxin, tumour necrosis factor-alpha and nitric oxide. AB - 1. Hyperdynamic circulation observed in portal hypertensive states is characterized by generalized vasodilation, increased cardiac index and increased systemic and regional blood flows. Endotoxin, tumour necrosis factor-alpha (TNF alpha) and nitric oxide (NO) have been reported to be involved in the pathogenesis of hyperdynamic circulation, but the interactions between endotoxin, TNF-alpha and NO in cirrhotic rats with ascites have never been specifically addressed. 2. This study was designed to determine systemic and portal haemodynamics and plasma levels of endotoxin, TNF-alpha and nitrate/nitrite in cirrhotic rats with ascites and investigate the relationships between these substances. 3. Plasma concentrations of endotoxin, TNF-alpha and nitrate/nitrite (an index of NO production) were determined in 25 cirrhotic rats with ascites and 17 control rats using the Limulus assay, ELISA and a colorimetric assay respectively. In addition, haemodynamic studies were performed in another ten cirrhotic rats with ascites and ten control rats. 4. Cirrhotic rats with ascites had hyperdynamic circulation accompanied by increased plasma levels of endotoxin, TNF-alpha and nitrate/nitrite, as compared with control rats. Significant correlation existed between plasma levels of endotoxin and nitrate/ nitrite (r = 0.59, P < 0.0001) and between plasma levels of endotoxin and TNF-alpha (r = 0.63, P < 0.0001). No correlation was detected between plasma levels of TNF-alpha and nitrate/nitrite (r = 0.24, P > 0.05). 5. This study suggests that endotoxaemia developed in cirrhotic rats with ascites may stimulate NO formation directly or indirectly via cytokine cascade, and consequently participate in the development and/or maintenance of hyperdynamic circulation. PMID- 9337637 TI - Nitric oxide and vascular reactivity in pregnant rats with adriamycin nephropathy. AB - 1. In previous studies we have shown that, after the administration of adriamycin, hypertension developed in rats who became pregnant (adriamycin pregnant rats), whereas virgin animals remained normotensive. Subsequently, we showed that this hypertension was prevented by administration of L-arginine, suggesting that deficient synthesis of nitric oxide may be pathogenetic in this model. 2. To further assess the role of nitric oxide in this model, we measured mean arterial blood pressure after administration of L-arginine to adriamycin pregnant rats or of NG-nitro-L-arginine-methyl ester (L-NAME) to normal pregnant rats. In other experiments, we assessed the response of isolated perfused arterial mesenteric vessels, precontracted with noradrenaline, to acetylcholine, L-arginine or L-NAME. 3. Blood pressure was decreased in normal pregnant rats, whereas it was elevated in adriamycin-pregnant rats. L-NAME treatment increased blood pressure in normal pregnant rats and L-arginine decreased it in adriamycin pregnant rats. 4. Mesenteric vessels of adriamycin-pregnant rats exhibited an exaggerated vasoconstrictory response to noradrenaline, when compared with the blunted response observed in normal pregnancy. The addition of L-NAME in vitro induced a further contraction, significantly greater in normal pregnant rats. The vasodilatory response to acetylcholine and L-arginine was greater in vessels from adriamycin-pregnant rats. In contrast, responses to either nitroprusside or diazoxide were similar in all groups. 5. The results suggest a state of reduced nitric oxide synthesis in rats with adriamycin nephropathy, leading to vascular maladaption and hypertension in pregnancy. PMID- 9337638 TI - Impaired insulin-induced attenuation of noradrenaline-mediated vasoconstriction in insulin-resistant obese Zucker rats. AB - 1. Insulin resistance is associated with hypertension but the underlying mechanism is unclear. We tested the hypothesis that insulin-induced vasodilatation is impaired in insulin-resistant obese Zucker rats. We studied mesenteric artery (approximately 220 microns diameter) function before the development of hypertension in 3-month old obese Zucker rats and age-matched lean rats. 2. In vessels from lean rats, insulin at concentrations of 50, 500 and 5000 m-units/l attenuated the constriction in response to noradrenaline (50 m-units/l: 8 +/- 3%, P < 0.05; 500 m-units/l: 13 +/- 3%, P < 0.02; 5000 m-units/l: 13 +/- 2%, P < 0.02). 3. Vessels from obese rats failed to show any such response to insulin (2 +/- 6% increase in maximal tension with 5000 m-units/l; not significant), both in the presence and absence of L-arginine (3 mmol/l). 4. Vessels from obese rats showed slight but significant impairment in the vasodilator response to acetylcholine (5 x 10(-8)-10(-4) mol/l) (obese: 64.1 +/- 3.7% relaxation; lean: 77.3 +/- 3.7% relaxation; P < 0.05); however, relaxation in response to A23187 was not significantly different between the phenotypes (obese: 81.3 +/- 10.6% relaxation; lean: 79.1 +/- 9.7% relaxation; not significant). 5. Systolic blood pressure was not significantly different in lean (126 +/- 8 mmHg) and obese (127 +/- 7 mmHg) rats at the time of study (not significant). 6. We conclude that insulin-induced attenuation of noradrenaline mediated vasoconstriction is impaired in the obese Zucker rat and that this defect precedes and therefore could contribute to the development of hypertension in this insulin-resistant model. The defect in insulin action could reside in the endothelial generation of nitric oxide, as endothelial function is also abnormal. PMID- 9337639 TI - Evidence for an R(+)-[(dihydroindenyl)oxy]alkanoic acid-sensitive K+/Cl- co transporter in human platelets and its interaction with the Na+/K+/2Cl- co transporter. AB - 1. The K+/Cl- co-transport system is activated by a number of interventions, such as cell swelling and stimulation with N-ethylmaleimide. It is specifically inhibited by R(+)-[(dihydroindenyl)oxy]alkanoic acid and requires the presence of K+ and Cl- on the same side of the cell membrane. This co-transporter has been studied extensively, mainly in erythrocytes of many species, in which it plays a key role in cell volume regulation. Here we present evidence that human platelets contain K+/Cl- co-transporters. 2. We have studied the efflux of 86Rb+ (a marker for K+) from 86Rb(+)-loaded human platelets, and have defined their response to stimulation by N-ethylmaleimide. 3. N-Ethylmaleimide (0.5 and 1 mmol/l) stimulated an increase in cumulative 86Rb+ efflux in a concentration-dependent manner. This efflux was inhibited by R(+)-[(dihydroindenyl)oxy]alkanoic acid (10 mumol/l) but was insensitive to bumetanide. It also required the presence of external Cl-. 4. These observations suggest that 86Rb+ efflux from the platelets stimulated by N-ethylmaleimide occurs via K+/Cl- co-transport. 5. When the K+/Cl- co-transporter was stimulated by N-ethylmaleimide we were unable to stimulate the Na+/K+/2Cl- co-transporter with a high external concentration of KCl or inhibit 86Rb+ efflux with bumetanide. Together with other evidence, this suggests that when the K+/Cl- co-transporter is stimulated with N-ethylmaleimide, the Na+/K+/2Cl- co-transporter is inhibited. PMID- 9337640 TI - Pharmacological evidence of calcium-activated and voltage-gated potassium channels in human platelets. AB - 1. Previous electrophysiological studies have suggested the presence of KCa and Kv channels in human platelets. However, the pharmacology of these channels has not been defined. 2. We have studied potassium channels in human platelets by measuring the efflux of 86Rb+ (a marker for K+) from 86Rb(+)-loaded cells, and have defined their responses to stimulation by the platelet agonist thrombin and the calcium ionophore ionomycin. 3. Thrombin (0.1-0.6 i.u./ml) stimulated an increase in 86Rb+ efflux from the platelets in a concentration-dependent manner. This efflux was significantly inhibited by apamin (100 nmol/l), charybdotoxin (300 nmol/l) and alpha-dendrotoxin (100-200 nmol/l), blockers of SKCa channels, KCh channels and Kv channels respectively. Iberiotoxin (300 nmol/l), a specific inhibitor of BKCa channels, had no effect on the thrombin-stimulated 86Rb+ efflux. Although glibenclamide, an inhibitor of KATP channels, inhibited the thrombin-stimulated efflux, it did so only in a high concentration (20 mumol/l). 4. Ionomycin (1-5 mumol/l) stimulated an increase in 86Rb+ efflux from the platelets in a concentration-dependent manner. This efflux was significantly inhibited by apamin (100 nmol/l) and charybdotoxin (300 nmol/l). However, iberiotoxin (300 nmol/l) had no effect on the ionomycin-stimulated 86Rb+ efflux. 5. These findings suggest that 86Rb+ efflux from platelets stimulated by thrombin and ionomycin occurs via two types of KCa channel: SKCa and KCh channels. Thrombin also stimulated efflux via Kv channels. PMID- 9337641 TI - Different dietary calcium intake and relative supersaturation of calcium oxalate in the urine of patients forming renal stones. AB - 1. Dietary calcium restriction, an efficient practice in reducing urinary calcium excretion, has been reported to induce either an increase or no change in oxalate excretion, questioning its use in hypercalciuric stone-forming patients. In addition, calcium restriction has been previously demonstrated to induce other urinary changes which might influence the relative supersaturation of calcium oxalate. So the overall effect of calcium deprivation on the relative supersaturation of calcium oxalate is unpredictable. 2. The aim of the study was to evaluate the effect of dietary calcium restriction on the relative supersaturation of calcium oxalate in the urine of stone-forming patients utilizing a computer methodology which takes into account the main soluble complex species of oxalate. 3. We studied 34 stone-forming patients on both a free-choice diet, whose Ca and oxalate content (24 and 1.2 mmol respectively) was assessed by dietary inquiry, and after 30 days on a prescribed low-calcium and normal oxalate diet (11 and 1.1 mmol respectively). Under both conditions, the excretion of the main urinary parameters related to dietary composition, electrolytes, oxalate and daily citrate urinary excretion, were measured. The relative supersaturation of calcium oxalate was calculated by means of an iterative computer method which takes into account the main soluble complex species on which the solubility of calcium oxalate is dependent. In addition, intact parathyroid hormone and 1,25-dihydroxyvitamin D blood levels were also evaluated. In 13 of the patients intestinal calcium absorption was evaluated during both a free- and a low-calcium diet, utilizing kinetics methodology. 4. The low-calcium diet induced, together with an expected reduction of calcium excretion, a marked increase in oxalate urinary output. This finding was independent of the presence or otherwise of hypercalciuria and of the serum levels of parathyroid hormone and vitamin D. Intestinal calcium absorption was also stimulated by calcium deprivation and its levels were well correlated with oxalate excretion. Minor changes in magnesium and citrate excretion were also observed. The overall effect on the relative supersaturation of calcium oxalate consisted in a substantial increase in this parameter during the low-calcium diet. 5. In conclusion, our data reinforce the concept that dietary calcium restriction has potentially deleterious effects on lithogenesis, by increasing the relative supersaturation of calcium oxalate. PMID- 9337642 TI - Enrichment in urinary ammonia and urea with hourly oral doses of [15N]glycine: evidence for a step function and a circadian rhythm in protein turnover. AB - 1. The present study sought to determine the possible existence of a pool of proteins which turn over with life-time kinetics. The pattern of enrichment of ammonia and urea in hourly samples of urine was determined in normal adults to whom oral doses of [15N]glycine were given hourly for 36 h. The subjects received hourly meals throughout, and in six the study commenced at 06.00 hours, in five at 12.00 hours and in two at 18.00 h. 2. A plateau level of enrichment was achieved in urinary ammonia within 4-6 h. Regardless of the time at which the study started this plateau was held until about midnight, at which time there was an increase in enrichment, with a second higher plateau 5-6 h later. The second plateau was held to the end of the study. For urinary urea the rate of rise in enrichment was slower and smoother, because of the slow turnover of the urea pool. 3. Protein synthesis, derived from the first ammonia plateau, 179 mg h-1 kg 1, was significantly higher than that derived from the second plateau, 118 mg h-1 kg-1. Using the plateau in urea towards the end of the 36 h, the estimate of protein synthesis was 153 mg h-1 kg-1. 4. The results are considered to provide evidence of a pool of proteins for which degradation takes place in harmony with a circadian rhythm. PMID- 9337643 TI - Plasma concentration of total leptin and human lung-cancer-associated cachexia. AB - 1. Adipocyte-derived leptin is postulated to represent the afferent hormonal signal to the hypothalamus in a feedback mechanism that regulates fat mass. In this proposed feedback mechanism, increased fat mass leads to an elevated plasma leptin level that eventually induces a decrease in appetite and an increase in energy expenditure, and vice versa. 2. As anorexia and hypermetabolism play a role in the development of cancer cachexia, we investigated the hypothesis that underlying abnormalities in the leptin feedback mechanism (in particular relatively high plasma leptin levels or, on the other hand, a hypothalamic insensitivity to a fall in leptin levels) might be involved. For this purpose, total plasma leptin, body composition (fat mass and fat-free mass), appetite and resting energy expenditure were assessed in 21 male lung-cancer patients. 3. Total leptin was detectable in six patients and non-detectable in 15. In comparison with the latter, the patients with detectable leptin were characterized by a trend towards less weight loss (3.4% compared with 11.0%, P = 0.07), as being less underweight (body mass index 23.8 kg/m2 compared with 19.4 kg/m2, P = 0.004) and by a higher fat mass (21.4 kg compared with 9.7 kg, P = 0.001). Significant between-group differences in appetite and resting energy expenditure were lacking. 4. Based on these findings, we conclude that in cancer the afferent part of the leptin feedback mechanism functions normally and that, in particular, elevated leptin levels are not involved in the development of cachexia. Since the absence of plasma leptin was not associated with an increased appetite and decreased energy expenditure, disturbances in the hypothalamic part of the feedback mechanism are hypothesized. PMID- 9337644 TI - Functional analysis of histamine release from basophils and mast cells in subjects with the Ile-181-->Leu variant of Fc epsilon RI-beta. AB - 1. Atopy is a genetically heterogeneous disorder, but there is now strong evidence that one important locus is on chromosome 11q13. Fc epsilon RI-beta at that location has been identified as a candidate gene and variants have been associated with atopy in population studies. 2. No information is available on the functional consequences of any of these variants, and defining this may prove difficult because of the complexity of the atopy phenotype and because Fc epsilon RI beta is expressed on a range of cells with different functions, including basophils, mast cells, eosinophils and antigen-presenting Langerhan's cells. 3. We have conducted a qualitative study of mast cell and basophil histamine release in nine atopic individuals with Ile-181-->Leu mutation of Fc epsilon RI beta, and ten unrelated similarly atopic individuals without Ile-181-->Leu mutation. There were non-significant trends for Ile-181-->Leu-positive atopic subjects to produce wheal responses at lower allergen challenge in skin prick tests, and to release more histamine from basophils following in vitro allergen challenge. 4. The data do not provide decisive evidence of functional differences between atopic subjects with Ile-181-->Leu and other atopic individuals; more discriminating functional experiments are required. PMID- 9337671 TI - Economic analyses of benefit from interferon-alpha 2B in high-risk melanoma: trade-offs between completeness, simplicity and clarity. PMID- 9337672 TI - Prognostic factors for advanced seminoma--a solid basis for clinical trials. PMID- 9337673 TI - Should high-dose chemotherapy be used in the treatment of soft tissue sarcoma? PMID- 9337674 TI - Phase II trials in the EORTC. The Protocol Review Committee, the Data Center, the Research and Treatment Division, and the New Drug Development Office. European Organization for Research and Treatment of Cancer. AB - The Protocol Review Committee (PRC) has prepared this document in order to clarify the phases of introduction of a new therapeutic agent in the treatment of cancer. Our aim is to define these phases and to make explicit the intentions which lie behind EORTC studies at each stage in the process. The PRC hopes that this will help the Cooperative Groups of the EORTC in developing their protocols for clinical trials carried out under the auspices of the EORTC. This position paper may also assist those undertaking this form of clinical research outside the EORTC. PMID- 9337675 TI - 5-HT3 receptor antagonists: differences and similarities. AB - Differences among 5-HT3 receptor antagonists have been reported in pharmacological studies with regard to selectivity of receptor binding, potency, duration of action and dose-response curves. However, whether these pharmacological differences can affect clinical efficacy and safety remains to be determined. A careful analysis of the literature revealed 22 comparative studies among the 5-HT3 receptor antagonists available for review. Unfortunately, several of these trials have some important shortcomings especially in the study design, the size of population studied and the type of anti-emetic treatment selected, making their conclusions often difficult to interpret. However, among these studies, seven large, double-blind clinical trials have clearly shown that the antiemetic activity and tolerability of ondansetron, granisetron, tropisetron and dolasetron is almost identical at least in the prevention of cisplatin-induced emesis. Therefore, from the efficacy and safety point of view, there is no reason to prefer one with respect to the other compound. From the economic perspective, instead, differences may exist and they are strictly related to the dose and schedule of administration chosen for each compound. The information available on the use of 5-HT3 receptor antagonists in the prevention of emesis induced by moderately emetogenic chemotherapy is at best scant. Contrasting results have been reported and only one well-conducted study has been published in full. Therefore, the possible differences among the various compounds are difficult to evaluate. More studies should be carried out in this group of patients. PMID- 9337676 TI - Cochrane cancer clan gathers in Brussels. PMID- 9337677 TI - A retrospective cost-effectiveness analysis of interferon as adjuvant therapy in high-risk resected cutaneous melanoma. AB - To assess the cost per life year gained of alpha interferon (IFN) as adjuvant therapy for patients with high-risk resected melanoma, we conducted a retrospective, incremental cost-effectiveness analysis on clinical data from a previously published ECOG trial [9]. The Gompertz model was used to estimate the total lifetime values of patient-years of subjects receiving IFN in comparison with subjects given no adjuvant treatment. The ECOG trial involved 143 patients treated with high-dose IFN and 137 given no adjuvant treatment. Estimated drug expenditures were based on the assumption of a cost of $109.25 per 10 MU of IFN. Our analysis of the ECOG results showed that the adjuvant treatment of 100 subjects with high-dose IFN improved survival expectancy by 133.6 discounted life years or 308 undiscounted life years. The use of IFN (compared with no adjuvant treatment) implied an incremental cost of $16,467 per discounted life year saved (95% CI of $4752-50,000) or $7143 per undiscounted life year saved (95% CI of $3226-33,846). Sensitivity testing, in which variations were introduced in the main factors influencing cost and effectiveness, showed that this value always remained below $50,000. Our pharmacoeconomic analysis indicates that adjuvant treatment with high-dose IFN in patients with high-risk resected melanoma implies a favourable cost-effectiveness ratio. Because two other studies showed no significant survival benefit in patients receiving adjuvant IFN at lower values of total dose per patient, the controversy remains and confirmation data are needed for the ECOG trial's results. If these clinical results are confirmed, our analysis shows that the dosage of IFN given in this trial has a favourable pharmacoeconomic profile. PMID- 9337678 TI - Prognostic factors for patients with advanced seminoma treated with platinum based chemotherapy. AB - Prognostic factors for 3-year progression-free survival (PFS) were defined in 286 patients with advanced seminoma treated with cisplatin-based chemotherapy at 10 European oncology units (no prior treatment: 236; prior radiotherapy: 50). Previously irradiated patients displayed a 69% PFS as compared to 87% in those presenting with advanced seminoma at the time of diagnosis (P = 0.009). In the univariate analysis, the extent and site of disease before chemotherapy and the level of serum LDH (< 2.0 versus > or = 2.0 x upper limit of normal) correlated with PFS in previously non-irradiated patients, but not in patients with prior radiotherapy. The multivariate analysis was, therefore, restricted to previously non-irradiated patients. The presence of non-pulmonary visceral metastases and a serum LDH level of > or = 2 x normal (N) proved to be independent prognostic factors. Based on these variables, two prognostic models were constructed and validated in an external data set of 166 comparable patients. For clinical use, Model 2 is recommended. The good-prognosis group comprises non-irradiated patients with stage II seminoma and any LDH level at presentation, or stage III and IV patients (with lung metastases only) whose serum LDH level is < 2 x N. These patients display a 94% 3-year PFS. The poor prognosis group includes all other patients with a 56% PFS. With this prognostic model, individualisation of the therapeutic approach may be considered in patients with advanced seminoma and a high risk of chemotherapy-related toxicity. PMID- 9337679 TI - Differential responses to chemoimmunotherapy in patients with metastatic malignant melanoma. AB - An open, multicentre non-randomised study was performed to evaluate the activity and toxicity of combination chemoimmunotherapy, consisting of cisplatin, interleukin-2 and interferon-alpha, in metastatic malignant melanoma. Between March 1992 and September 1993, 28 patients with pathologically proven metastatic malignant melanoma, bidimensionally measurable disease and an Eastern Co operative Oncology Group score < or = 1 were treated with the combination chemoimmunotherapy. The regimen consisted of cisplatin (100 mg/m2 on day 0), interleukin-2 (Proleukin, Chiron, Middlesex, U.K.) 18 x 10(6)IU/m2/d continuous intravenous infusion on days 3-7 and 17-22, with interferon-alpha (Roferon-A, Roche, Hertfordshire, U.K.) 9 x 10(6) U/d subcutaneously on days 3, 5, 7, 17, 19, 21 during the interleukin-2 infusions. The treatment cycle lasted 28 days. Among 27 assessable patients, 5 patients achieved partial responses, for an overall response rate of 18% (95% CI 6-37%). Median progression-free survival was 44 days (range 8-279) and median overall survival was 264 days (range 41-1432). Differential responses were noted in 41% of patients and responses were more frequent in non-visceral disease (skin, lymph node and soft tissue disease) (P = 0.04). These results indicate that differential responses to chemoimmunotherapy are common in patients with metastatic melanoma. This may account for the broad range of response rates reported in the literature. PMID- 9337680 TI - Neurological monitoring of neurotoxicity induced by paclitaxel/cisplatin chemotherapy. AB - To evaluate the neurotoxicity of paclitaxel/cisplatin chemotherapy, we studied neurological and electrophysiological functions in 14 patients who had been treated with 1-7 courses of paclitaxel/cisplatin. The cumulative paclitaxel and cisplatin doses ranged from 175 to 1225 mg/m2 and 100-700 mg/m2, respectively. Neurological examinations as well as motor nerve conduction studies of the peroneal nerve were performed and summarised by means of a peripheral neuropathy score. Neurotoxicity with onset usually after the second treatment cycle occurred in 13 patients. 12 patients complained about sensory symptoms, 13 patients had impaired vibration sense and 8 patients developed additional muscle weakness, predominantly of the legs. Dysfunction of peroneal motor nerve conduction occurred in 13 patients. Reduction of amplitudes as well as slowing of conduction velocities were seen in 13 patients and prolonged distal latencies in 10 patients. The peripheral neuropathy score was elevated in 13 patients. Neurological symptoms, impairment of both vibration sense and tendon reflexes, and the peripheral neuropathy score increased with the cumulative doses of paclitaxel/cisplatin. Serial analysis among selected patients also revealed an increase in neurotoxicity with increasing cumulative drug doses. These data indicate the development of neurotoxicity in most patients treated with paclitaxel/cisplatin and also suggest that early signs of neurotoxicity can be detected by clinical examination with emphasis on symptoms as well as vibration sense and can be well documented by electrophysiological investigations. PMID- 9337681 TI - Oxaliplatin/cisplatin (L-OHP/CDDP) combination in heavily pretreated ovarian cancer. AB - The aim of this study was to evaluate the toxicity and the activity of two non cross-resistant platinum compounds: oxaliplatin (L-OHP) and cisplatin (CDDP) in platinum pretreated ovarian cancer patients. Chemotherapy consisted of L-OHP and CDDP given sequentially as 2 h infusions on day 1 at their standard recommended dose (130 mg/m2 for oxaliplatin, 100 mg/m2 for cisplatin) every 3 weeks. Dose reductions (20-35%) were planned according to baseline haematological and renal status, but the dose ratio between L-OHP and CDDP was always maintained at 1.3. Cycles were repeated until progression or treatment limiting toxicities. From September 1992 to November 1994, 25 patients with pretreated ovarian cancer entered this salvage programme. They had received a median number of three previous chemotherapy lines (1-7), one at least platinum based. Previously cisplatin had been given to 22 patients at a median total dose of 600 mg/m2 (170 1175), while 18 had received carboplatin to a median total dose of 1135 mg/m2 (200-2450). 9 patients had also received and were resistant to taxanes (paclitaxel, 6 patients, docetaxel, 3 patients), while the rest were considered ineligible for simultaneously ongoing single-agent taxane phase II trials. 13 and 12 patients, respectively, were considered to have platinum refractory and potentially sensitive disease, according to Markman's criteria. 77 cycles of L OHP/CDDP were given, with a median of three cycles/patient (range 1-6) and were evaluable for toxicity. The limiting toxicity of the L-OHP/CDDP combination was a cumulative, sensory peripheral neuropathy, severe (> or = grade 3 CTC) after more than three cycles, but reversible within a few months of its discontinuation. Grade 3-4 (WHO scale) neutropenia and thrombopenia were seen in 35-40% of cycles, with one neutropenic treatment-related death (septic shock). 22 patients with measurable/evaluable disease were assessable for antitumoral activity. Two complete responses (CR) (8%) (one proven histologically at laparotomy (pCR)) and 8 partial responses (PR) (32%) for an overall objective response rate (ORR) of 40% (95% CI, 21-61%) (intent to treat). The median duration of response was 4 months. Seven responses were seen among 12 potentially platinum-sensitive tumours (58%, CI 95% 28-85%), while 3/13 platinum refractory patients (23%, CI 95% 5-54%) had an objective response. These encouraging results are the basis for new first- and second-line combination treatment programmes in ovarian carcinoma. PMID- 9337682 TI - Predictive value of serum medroxyprogesterone acetate concentration for response in advanced or recurrent breast cancer. AB - Medroxyprogesterone acetate (MPA) is one of the most commonly prescribed drugs for endocrine therapy of metastatic breast cancer. In this study, the serum MPA concentration was measured by high-performance liquid chromatography (HPLC) and evaluated for its usefulness in predicting the response in 79 cases of advanced or recurrent breast cancers. Overall, 29 patients (37%) achieved an objective response. The response rate correlated significantly with the oestrogen receptor (ER) status (P = 0.03), proliferative activity determined by DNA polymerase alpha (P = 0.04), the disease-free interval (DFI) (P = 0.05) and the serum MPA concentration (P < 0.001). Patients with ER-positive tumours, lower proliferative activity, a longer (DFI) or a higher serum MPA concentration responded more frequently. The mean serum MPA concentration in the responders with ER-positive tumours (P = 0.01) or tumours with a lower proliferative activity (P = 0.008) were significantly lower than in cases with ER-negative tumours or tumours with a higher proliferative activity, respectively. Cases with soft tissue metastases showed responses at significantly lower MPA concentrations (P = 0.003) than those with bone or visceral metastases. Furthermore, there was a dramatic decrease in the MPA concentration when a responder with a high concentration became unresponsive to the therapy. Thus, the serum MPA concentration is a determining factor for the response to treatment. PMID- 9337683 TI - Carcinoma of the stomach following the Chernobyl nuclear accident. AB - Medical consequences of many nuclear accidents on humans are well studied, but the results pertaining to gastric cancer patients who were exposed to radiation as a result of the Chernobyl nuclear accident have not been analysed. In this study, the outcome of the surgical treatment of 68 gastric cancer patients who were exposed to radiation as a result of the Chernobyl nuclear accident was compared with that of 117 consecutive gastric cancer patients from uncontaminated areas of the Ukraine. Patients in the study group was significantly younger than that of the control group. Comparative analysis showed the same frequency of regional metastases (65.7% versus 71.1%, P > 0.05), but a smaller number of distant metastases (23.8% versus 38.1%, P < 0.05) in the study group. 41.2% of patients in the study group underwent total gastrectomy compared to 19.6% of patients in the control group (P = 0.002). Postoperative complications developed in 13.2% of patients in the study group, while postoperative mortality in the study group was 7.3% compared to 1.7% in the control group. A significant decrease in CD16 cells was noted in patients from the study group following the operative procedure. Young age, invasive tumours with smaller number of distant metastases, frequent necessity for total gastrectomy and combined operations with adjacent organs, a higher level of postoperative morbidity and mortality and low levels of natural killer cells (CD16+) with a tendency to decrease after surgery are characteristic of patients with carcinoma of the stomach affected by the Chernobyl accident. PMID- 9337684 TI - Paclitaxel changes sympathetic control of blood pressure. AB - Paclitaxel has become part of standard therapy in the treatment of ovarian and breast cancer. Concern has been raised about the effects of paclitaxel on cardiovascular function. Therefore, this study of the effects of paclitaxel on autonomic cardiovascular control was initiated. Eighteen women treated for ovarian or breast cancer were examined with autonomic cardiovascular function tests, once before the treatment and once after the second course of paclitaxel. Heart rate and blood pressure variability and changes in heart rate and blood pressure responses to the tests were measured. Baroreflex sensitivity was calculated from the Valsalva manoeuvre non-invasively. Paclitaxel did not change heart rate variability at rest compared with the pretreatment level. However, medium frequency variability of blood pressure was smaller after treatment with paclitaxel. Paclitaxel treatment did not impair the heart rate and blood pressure responses to the autonomic function tests. The results do imply that paclitaxel alters sympathetic control of blood pressure. Nevertheless, paclitaxel does not appear to precipitate autonomic cardiac neuropathy. PMID- 9337685 TI - Pharmacokinetics of amifostine and its metabolites in patients. AB - The pharmacokinetics of the cytoprotective agent amifostine (EthyolR; WR 2721) and its main metabolites (WR 1065 and the disulphides) were studied in patients participating in two phase I trials concerning carboplatin or cisplatin in combination with amifostine. Patients were treated with a single dose or three doses of amifostine (740 or 910 mg/m2). The single or first dose was given as a 15 min i.v. infusion just before administration of the chemotherapeutic agent. The additional two infusions were administered 2 and 4 h thereafter. Amifostine was rapidly cleared from the plasma, due to, at least in part, the fast conversion into WR 1065. A biphasic decrease with a final half-life of 0.8 h was observed. The active metabolite WR 1065 was cleared from the plasma with a final half-life of 7.3 +/- 3.6 h. The short initial half-life of WR 1065 can be explained by its fast uptake in tissues and the formation of disulphides. The disulphides were cleared with a final half-life of 8.4-13.4 h and were detectable for at least 24 h after treatment. They may serve as an exchangeable pool of WR 1065. The amifostine peak values at the end of each 15 min infusion did not accumulate in the multiple dosing schedule. For WR 1065 a trend towards an increase in the peak levels was observed [C1,max: 47.5 +/- 11.9 microM, C2,max: 79.0 +/- 13.2 microM, C3,max: 84.8 +/- 15.1 microM, (n = 6)], whereas a trend towards a small decrease was observed for the peak levels of the disulphides [C1,max: 184.2 +/- 12.6 microM, C2,max: 175.0 +/- 23.7 microM, C3,max: 166.0 +/- 17.2 microM, (n = 6)]. This latter finding might suggest a saturation of the disulphide formation or a change in the uptake or elimination of WR 1065, which would result in higher WR 1065 levels in plasma and tissues, after multiple doses of amifostine. PMID- 9337686 TI - Neuroblastoma. AB - The neuroblastic tumours, derived from primordial neural crest cells which ultimately populate the sympathetic ganglia, adrenal medulla and other sites, (Brodeur GM and Castleberry RP. Neuroblastoma. In Pizzo PA, Poplack DG, eds, Principles and Practice of Pediatric Oncology. Philadelphia, J. B. Lippincott Co., 1997, 761-797) are an enigmatic group of neoplasms which have the highest rate of spontaneous regression of all human malignant neoplasms yet one of the poorest outcomes when occurring as disseminated disease in children. Significant advances in understanding and predicting the natural history of neuroblastoma have resulted from translational studies coupling tumour biology and clinical features to form prognostic strata and allowing more accurate routeing of patients to risk-related management. While this strategy has clarified the management for lower risk tumours, little improvement in survival for higher risk disease has been realised. Ironically, this latter patient subset, for which the most innovative therapeutic strategies are needed, is also the one from which the least tumour biology is gleaned owing to inadequate tissue sampling. This update will summarise the evolving biology of neuroblastoma and its relationship to current risk-related therapy and future management strategies. Throughout this report, prognostic grouping by age will be infants (< 1 year) versus children (> or = 1 year) since the change of risk according to age seems most distinct at this cut-off point. PMID- 9337687 TI - The U.S. pediatric cancer clinical trials programmes: international implications and the way forward. AB - The four national paediatric cancer clinical trials organisations in the United States--the Children's Cancer Group, the National Wilms' Tumor Study Group, the Intergroup Rhabdomyosarcoma Study Group and the Pediatric Oncology Group--were formed in 1955, 1969, 1972 and 1979, respectively. Together, the Children's Cancer Group and Pediatric Oncology Group serve as a national registry of nearly all childhood cancers in the United States, provide a national network of communication for researchers, care providers and families of paediatric patients with malignant disease and conduct laboratory investigations and clinical trials of new treatments of cancers in infants, children, adolescents and young adults. Nearly 95% of patients with cancer in the United States who are below 15 years of age are registered by the Children's Cancer Group and the Pediatric Oncology Group and more than half of American children with cancer are entered into at least one trial by a paediatric group. Improved survival of children receiving treatment according to well-defined protocols in specialised children's centres, in contrast to children who received treatment outside of these centres, has been shown for those with acute lymphoblastic leukaemia, lymphoma, Wilms' tumour, medulloblastoma, rhabdomyosarcoma and Ewing's sarcoma. By the year 2000, the overall cure rate for United States children and adolescents with cancer should exceed 85%. To reach this goal, the way forward will depend on international collaboration, implementation of global harmonisation, prevention of the erosion of biomedical research and clinical trials by the managed health care industry, increased public and private financial support and continued recruitment into paediatric oncology of brilliant and dedicated young investigators. The specific challenges ahead include: (1) transferring the knowledge, methodologies and technologies to countries that are less fortunate; (2) conducting multinational clinical trials in conjunction with paediatric cooperative groups in other countries; (3) accessing older adolescent patients who currently do not participate in cooperative group trials; (4) merging clinical trials by adult collaborative groups that overlap with the paediatric groups, as in acute lymphoblastic leukaemia, acute myelogenous leukaemia, Hodgkin's disease, osteosarcoma and germ cell tumours; (5) establishing a stable source of funding for national and international cooperative paediatric cancer clinical trials; (6) creating an informatics system that can link paediatric cooperative group operation centres around the world, and the institutions within each collaborative group; and (7) securing the support of the insurance industry and government in covering clinical trials. PMID- 9337688 TI - The United Kingdom Children's Cancer Study Group--the first 20 years of growth and development. PMID- 9337689 TI - Interval cancers and cancers in non-attenders in the Ostergotland Mammographic Screening Programme. Duration between screening and diagnosis, S-phase fraction and distant recurrence. AB - The study was based on a population mammographic screening programme for women aged 40-74 years. Metastatic potential was analysed in 843 invasive breast cancers with regard to mode of detection and a number of prognostic factors. There was a higher metastatic capacity in clinically detected cases, but multivariate analyses showed that neither the mode of detection (hazard rate ratio of distant recurrence RR = 1.39, 95% CI 0.78-2.46 interval cancers and RR = 1.6, 95% CI 0.76-3.36 non-attenders) nor the duration between screening and diagnosis for true interval cancers (RR = 0.47, 95% CI 0.16-1.35 in tumours detected later than one year after screening) were independent prognostic factors. A correlation was found between metastatic potential and the SPF (RR = 2.94, 95% CI 1.57-5.50 in tumours with a high SPF), the oestrogen receptor status and the tumour stage. In conclusion, interval cancers intrinsically are not different from other breast cancers with equivalent characteristics; the duration between screening and diagnosis in interval cancers was not clearly correlated to the prognosis, but the S-phase fraction was a powerful predictor of prognosis. PMID- 9337690 TI - Duration of colorectal cancer symptoms and survival: the effect of confounding clinical and pathological variables. AB - The relationship between symptom duration and long-term survival following colorectal cancer is complex, and a number of factors may influence the length of time from onset of symptoms of cancer diagnosis. We prospectively studied 777 consecutive colorectal cancer patients to determine the association between symptom duration and survival independent of other clinical and pathological features. We used survival curves, the logrank test and Cox's proportional hazards model to assess possible changes in relative risk of death with increasing symptom duration, without making any a priori assumptions. We found that symptom duration shortened with advanced tumour stage (P < 0.0006) and was also shorter for patients presenting with bowel obstruction (P < 0.0001). Univariate survival analysis showed that long-term survival increased consistently with symptom duration (P < 0.001). However, when the effect of tumour stage and bowel obstruction were accounted for in a multivariate analysis, no decrease in the relative risk of death was seen as symptom duration increased. The addition of other variables to the proportional hazards model such as age, sex or tumour site did not further influence the risk function form of symptom duration. Our results suggest that early diagnosis of colorectal cancer should remain our goal when assessing patients with suggestive gastrointestinal symptoms. PMID- 9337691 TI - Immunoneutralisation of gonadotrophin releasing hormone: a potential treatment for oestrogen-dependent breast cancer. AB - The aim of this study was to assess the therapeutic potential of active immunisation with GnRH-glycys-PPD in a hormone-dependent experimental model. Mammary tumours were induced in female rats using dimethylbenzanthracene (DMBA) and the effects of GnRH immunoneutralisation on tumour development were evaluated. High titres of anti-GnRH IgG correlated with a decrease in oestrogen levels and subsequent tumour suppression. A comparison of immunised and non immunised animals showed that when GnRH-specific IgG levels were at a maximum titre (80-100 micrograms/ml), nearly 10% of the GnRH-glycys-PPD treated animals showed mammary masses, compared with all the non-treated animals at the same stage in the study. When the antibody levels fell, tumour regrowth was observed, but to a level below that observed in the non-treated animals. Following further treatment with the analogue, the tumours regressed again, showing their retention of hormone dependency. This is consistent with other endocrine manipulations in the treatment of breast cancer; the advantages of immunisation with GnRH-glycys lies in its non-toxicity and reduction in side-effects, which were mainly adjuvant-induced. PMID- 9337692 TI - An intronic deletion in TP53 gene causes exon 6 skipping in breast cancer. AB - Six hundred and thirty primary breast cancer were screened for abnormalities in exons 5, 6, 7 and 8 of the TP53 tumour suppressor gene. Analysis of the structure of the TP53 gene exons was performed with the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method and with direct sequencing of amplified DNA. In a breast tumour case from a postmenopausal patient, we found a deletion of 36 bp in intron 5 and no immunohistochemical staining for p53. We amplified and sequenced the cDNA region between exons 4 and 7 and showed that the deletion causes the skipping of exon 6. The resulting mRNA sequence had a frameshift that yields an inactive protein with a truncated C terminus. These results show the first example of intronic deletion causing exon skipping at the TP53 gene level. PMID- 9337693 TI - Differential binding of nuclear proteins to the TP53 gene promoter in male breast tumour. AB - It is well established that TP53 regulates the expression of many genes, but the regulation of expression of TP53 itself is poorly understood. Recently, it has been shown that there is a tissue-specific binding of nuclear proteins in the TP53 gene promoter. The aim of this study was to determine the nuclear proteins that bind to the TP53 promoter elements (between -104 and -458) in male breast cancer. The results of our study, using the electrophoretic mobility shift assay (EMSA) and Southwestern analysis, have showed: (1) nuclear proteins or factors other than p53 bind to the TP53 promoter; (2) the levels of at least four nuclear proteins vary between normal and tumour breast tissue; and (3) two newly discovered nuclear proteins bind to the TP53 promoter in tumour tissue but are absent in normal tissue. This differential binding of nuclear proteins to the TP53 gene promoter might play a critical role in TP53 transcription and cancer progression in male breast tumours. PMID- 9337694 TI - Quantification of additional short arms of chromosome 12 in germ cell tumours using the polymerase chain reaction. AB - Male germ cell tumours are characterised by the over-representation of 12p sequences, most often in the form of isochromosome i(12p). This study describes the development of a quantitative detection system for additional copies of 12p employing the polymerase chain reaction (PCR). The validity of this method was assessed on two i(12p) containing tumour cell lines in which the number of i(12p) was determined by fluorescence in situ hybridisation. Fourteen primary male germ cell tumours were analysed using the PCR-based method. While 3/8 seminomatous germ cell cancers did not contain any additional 12p, all 6 non-seminomatous tumours did and the severity of the disease correlated with the respective copy number. The ease of the PCR-based method makes it possible for the quantification of additional 12p to become a routine diagnostic and prognostic tool for testicular germ cell tumours, thereby helping to define the role of the i(12p) anomality in larger retrospective studies. PMID- 9337695 TI - Game-theory models of interactions between tumour cells. AB - The population of cells that comprises a tumour may consist of genetically different individuals. Often, such polymorphisms result from the expansion of a new, advantageous clone. The hypothesis is presented that sometimes tumour cells may adopt genetically-determined strategies to boost their own replication at the expense of other cells in the tumour. Simple game-theory models have been used to study this hypothesis, taking as an example the hypothetical advantage gained by tumour cells which produce a cytotoxin to harm other tumour cells. The models show that genotypes which cause cells to produce cytotoxic substances can spread through the tumour cell population, as can genotypes for resistance to the cytotoxin; in other circumstances, stable polymorphisms between these strategies can occur. The path of the tumour cell population to internal or external equilibrium is often complex, with large fluctuations in genotype frequencies. Flexible strategies are usually superior to fixed strategies. As in populations of whole organisms, 'social' interactions between tumour cells can act in favour of the individual cell at the expense of the tumour as a whole: strategies that retard the growth of the tumour can be selected and tumour regression is theoretically possible. Many mutations in tumours, especially in large or late stage lesions, may play a role in relations between tumour cells rather than providing those cells with a simple replicative advantage. PMID- 9337696 TI - Messenger ribonucleic acid expression of LH/hCG receptor gene in human ovarian carcinomas. AB - The mRNA expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptors was analysed by the RT-nested PCR method in five normal ovarian tissues, 62 ovarian tumours (5 benign, 7 borderline and 43 malignant epithelial tumours, 3 sex cord-stromal tumours and 4 germ cell tumours) and in 2 ovarian cancer cell lines. In normal ovaries, two cDNA fragments of different sizes were detected using primers designed to amplify a region including exon 9. Sequencing revealed that the larger fragment was derived from a full-length receptor, while the smaller fragment was a splice variant lacking exon 9. In ovarian tumours, the larger fragment of LH/hCG receptors was detected in 40% of the epithelial ovarian carcinomas, none of the germ cell tumours, all of the sex cord-stromal tumours and one of the 2 ovarian cancer cell lines. Immunohistochemistry confirmed the localisation of LH/hCG receptor protein in the tumour cells which correlated with mRNA expression. Patients with full-length LH/hCG receptors in carcinomas showed a better prognosis compared with those without the receptors. PMID- 9337697 TI - Cytosolic phospholipase A2, cyclo-oxygenases and arachidonate in human stomach tumours. AB - Human stomach tumours usually form more prostaglandins (PGs) than their associated normal mucosa/submucosa, but the mechanisms are not fully understood. The key enzymes are cytosolic phospholipase A2 (cPLA2, Mr 85,000) and the cyclo oxygenases (COXs) which exist in constitutive (COX-1) and inducible forms (COX 2). In human stomach tumours and associated macroscopically normal tissues, we determined the fatty acid composition by gas chromatography, amounts of cPLA2, COX-1 and COX-2 by immunoblotting with specific antibodies and cPLA2 enzyme activity using a tritiated substrate. Although compared to normal mucosa there was less arachidonate in tumours (P < 0.05), the arachidonate/total fatty acid ratio was higher. Mean amounts of cPLA2 and COX-1 and cPLA2 activity were similar in tumours and normal mucosa. However, substantial amounts of COX-2 were found in the tumours but not in the mucosa, which may explain why many gastric tumours form increased amounts of PGs. PMID- 9337698 TI - Phase II study of weekly 4'-epidoxorubicin in patients with metastatic adenocarcinoma of the cervix: an EORTC Gynaecological Cancer Cooperative Group study. AB - In this study 22 patients with metastatic adenocarcinoma of the cervix were treated with a weekly bolus injection of 4'-epidoxorubicin at a dose of 12 mg/m2. Seventeen patients had received prior radiotherapy, all patients were chemo naive. Toxicity was generally absent or very mild. One patient had a complete response and 2 patients had a partial response, one was an unconfirmed partial response, giving a response rate of 14%. Six patients had stable disease. The median progression-free survival and overall survival was 2.8 months and 6.1 months, respectively. In conclusion, 4'-epidoxorubicin used at this dosage and schedule has minimal activity in metastatic adenocarcinoma of the cervix. PMID- 9337699 TI - MDR1 RNA transcripts do not indicate long-term prognosis in colorectal carcinomas. AB - Because P-glycoprotein expression might be associated with a more aggressive behaviour of colorectal carcinomas (Weinstein et al., Cancer Res, 1991, 51, 2720 2726), we determined the relationship between MDR1 RNA expression of the carcinomas and the survival of the patients. At a median duration of follow-up of 86 months, event-free survival of patients with MDR1 RNA-negative tumours (n = 35) was not significantly different to that of patients with MDR1 RNA positive tumours (n = 67). Among the different tumour stages, event-free survival of the patients was also independent of MDR1 gene expression of the tumours. Thus, these findings do not support the hypothesis that local aggressiveness of P glycoprotein positive tumour cells translates into worse clinical outcome. PMID- 9337700 TI - Remarkable age-dependent sex differences in the incidence of adenocarcinoma of the gastric cardia and oesophagus in The Netherlands. PMID- 9337701 TI - Artifactual hypoglycaemia during treatment with filgrastim (rHu-met-G-CSF) PMID- 9337752 TI - Low-dose combination therapy in hypertension. PMID- 9337753 TI - Death certificates: why it matters how your patient died. PMID- 9337754 TI - Pathologic gambling: America's newest addiction? PMID- 9337755 TI - Administering topical steroids in sinusitis with head inverted. PMID- 9337756 TI - How timely is AFP? PMID- 9337757 TI - Three types of chronic pain. PMID- 9337758 TI - Issues in the management of bacterial meningitis. AB - Acute bacterial meningitis is associated with significant morbidity and mortality despite the availability of effective antimicrobial therapy. The emergence of antibiotic-resistant bacterial strains in recent years has necessitated the development of new strategies for empiric antimicrobial therapy for bacterial meningitis. Specifically, the emergence of strains of Streptococcus pneumoniae that are resistant to penicillin and the cephalosporins have led to empiric therapy for patients with pneumococcal meningitis consisting of vancomycin plus a third-generation cephalosporin pending susceptibility testing. Third-generation cephalosporins are also effective as empiric therapy against other pathogens that cause community-acquired bacterial meningitis, with the exception of Listeria monocytogenes, for which ampicillin or penicillin G is the antimicrobial agent of choice. Adjunctive dexamethasone should be administered to infants and children with suspected or proven Haemophilus influenzae type b meningitis to reduce audiologic and neurologic sequelae; administration concomitant with or just before the first dose of the antimicrobial agent is optimal for best results. PMID- 9337759 TI - Milk, eggs and peanuts: food allergies in children. AB - True food allergies are much less prevalent than is generally believed. They are more common in infants and children under age three than in older children and adults. Infant colic generally is not caused by a food allergy. In infants, urticaria, eczema or gastrointestinal bleeding may be due to foods such as milk and eggs, but clinical tolerance usually develops within a few years. Peanuts, tree nuts, seafood and seeds, as well as milk and eggs, can cause anaphylaxis in highly allergic children, and reexposure to such foods presents the risk of life threatening reactions. Immediate-reacting allergy skin tests and in vitro IgE antibody tests can be used to screen for food allergy. Only food challenge, however, can confirm a reaction to a particular food. Management of food allergy, once the initial symptoms are confirmed, consists of avoidance of specific foods, sometimes for a lifetime. All children at risk for food anaphylaxis should be identified, and their parents or caretakers should be prepared to administer epinephrine before taking the child to the emergency room. PMID- 9337760 TI - Evaluation and treatment of orthostatic hypotension. AB - Orthostatic hypotension is defined as a decrease of at least 20 mm Hg in systolic blood pressure when an individual moves from a supine position to a standing position. Nonneurogenic causes of orthostatic hypotension are related to cardiac pump failure, reduced intravascular volume, venous pooling or a medication side effect. Neurogenic causes include both central and peripheral nervous system lesions. The diagnostic evaluation requires a systematic review of medications and coexisting medical conditions along with a neurologic examination to search for treatable factors that may be contributing to orthostatic hypotension. Specific testing of autonomic function is useful for detecting subclinical orthostatic hypotension or for monitoring autonomic function over a period of time. Treatment is directed at improving the patient's symptoms rather than achieving arbitrary blood pressure goals. PMID- 9337761 TI - 1997 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with HIV: Part III. Prevention of disease recurrence. U.S. Department of Health and Human Services, Public Health Service. Centers for Disease Control and Prevention. PMID- 9337762 TI - Chronic cough. AB - Chronic cough is defined as a cough that lasts for more than three weeks. More than 90 percent of cases of chronic cough result from five common causes: smoking, post-nasal drip, asthma, gastroesophageal reflux and chronic bronchitis. Although in most patients chronic cough has a single cause, in up to one fourth of patients, multiple disorders contribute to the cough. A stepwise evaluation in patients with chronic cough can minimize the invasiveness and expense of the work up. Initial screening of patients with chronic cough should search for smoking, occupational exposure to an airway irritant, cough-inducing medications, airway hyperresponsiveness following upper respiratory infection, chronic bronchitis or any systemic symptoms suspicious for serious disease. Patients who are not diagnosed after an initial screening are evaluated or empirically treated in a stepwise fashion for postnasal drip, asthma and reflux. Bronchoscopy is reserved for use in the few patients still without a diagnosis after the previous steps have been completed. PMID- 9337763 TI - Myocardial revascularization in 1997: angioplasty versus bypass surgery. AB - In patients with coronary artery disease and severe ischemia, angioplasty and coronary artery bypass surgery have been shown to reduce symptoms, improve functional capacity and, in some patients, prolong life. Six major randomized trials have recently been reported comparing bypass surgery with angioplasty in patients with multivessel coronary disease. Uniformly, these studies demonstrate an equal mortality and reinfarction rate over five years of follow-up. Patients with angioplasties needed a repeat procedure during follow-up far more frequently than patients with bypass needed an additional bypass procedure (30 to 40 percent versus 5 to 10 percent). Although angioplasty was initially less costly, over five years the costs for the two procedures were similar. Mortality rates decreased by twofold when patients with diabetes mellitus were treated with bypass surgery rather than angioplasty. These studies confirm that in nondiabetic patients, bypass surgery and angioplasty are equally effective in the treatment of severe coronary disease. In diabetic patients with severe disease, however, bypass surgery is favored. PMID- 9337764 TI - Melatonin. AB - Melatonin, a hormone produced by the pineal gland, appears to help regulate the sleep-wake cycle. With further study and clinical experience, it may become an accepted therapy for insomnia. Although melatonin preparations are available without prescription in health food stores and pharmacies, their potency, purity, safety and effectiveness cannot be assured. Until large clinical trials provide further information about melatonin's efficacy, adverse effects, drug interactions and effects on various disease states, melatonin products should be used with the understanding that many questions about their safety remain unanswered. PMID- 9337766 TI - American College of Cardiology and American Heart Association address the use of echocardiography. PMID- 9337765 TI - Certification of death by family physicians. AB - Death certificates are important because morbidity and mortality statistics often come from death-certification data. These statistics are vital in developing approaches to disease treatment and strategies for increasing longevity. It is often the primary care physician who is responsible for completing the death certificate, for explaining the cause of death to the family and, if appropriate, for referring some cases to the medical examiner. The primary care physician should have an explicit understanding of how to determine the cause and manner of death and should use succinct, clear language in completing the death certificate. When doubt exists or an external cause of death is a possibility, the coroner or the medical examiner is the appropriate public health official to contact. PMID- 9337767 TI - World Wide Web resources for family physicians. PMID- 9337768 TI - Childhood-onset schizophrenia. Progressive ventricular change during adolescence. AB - BACKGROUND: There is controversy about progression in brain abnormalities in later-onset schizophrenia. This study looked for more striking progression in brain abnormalities during adolescence in a chronically ill, treatment-refractory sample of patients with childhood-onset schizophrenia who had had more prepsychotic developmental disturbance, but clinical and neurobiological characteristics similar to those of patients with treatment-refractory adult onset schizophrenia who have poor outcome. METHODS: Anatomic brain magnetic resonance images were obtained for 16 children and adolescents with onset of schizophrenia by 12 years of age and 24 temporally yoked, age- and sex-matched healthy controls. Subjects were scanned on initial admission and rescanned after 2 years with the identical equipment and measurement methods. RESULTS: Childhood schizophrenics showed a significantly greater increase in ventricular volume than did controls, for whom ventricles did not increase significantly (analysis of variance, diagnosis x time, F = 16.1, P < .001). A significant decrease in midsagittal thalamic area was also seen for the schizophrenics (P = .03), which was unchanged at rescan for controls. These differential brain changes correlated significantly with each other and tended to be predicted by both prepsychotic developmental abnormality (Premorbid Assessment Scale, P = .06) and Brief Psychiatric Rating Scale at follow-up (P = .07). CONCLUSIONS: More consistent progressive ventricular enlargement was seen during adolescence for this childhood-onset sample than has been reported for adult-onset populations. The brain imaging results support other clinical data showing both early and late deviations in brain development for at least this rare subgroup of treatment refractory, very-early-onset schizophrenic patients. PMID- 9337769 TI - Autonomic nervous system markers of psychopathology in childhood-onset schizophrenia. AB - BACKGROUND: Consistent abnormalities in peripheral indicators of autonomic activity, ie, skin conductance (SC) and heart rate (HR), have been reported in adult-onset schizophrenia. Herein, we use these markers to test the hypothesis of continuity between childhood-onset schizophrenia and adult-onset schizophrenia. METHODS: Skin conductance and HR were recorded from 21 severely ill children and adolescents (mean age, 14.1 years) with childhood-onset (< or = 12 years) schizophrenia (patient group) and from 54 age-matched controls (control group) during a rest period, a series of innocuous tones, reaction time instructions, and a simple warned reaction time task. RESULTS: During rest, patients had higher rates of spontaneous SC responses (SCRs) and HRs than controls, but their SC level was marginally lower and declined more slowly over time. Half of the patients, compared with 4% of the controls, failed to give SC-orienting responses to the first 2 tones. Patients who responded had impaired SCR magnitudes, and their habituation was more erratic than that of controls. The increase in SC level and SCR frequency at the onset of the task period was greatly attenuated in the patients, so that both variables were higher in controls. Patients had smaller SCRs and anticipatory HR responses to the reaction time stimuli. Skin conductance nonresponding was associated with negative and total symptoms, and spontaneous SCR frequency was associated with positive symptoms. CONCLUSIONS: The findings show similar abnormalities in autonomic nervous system activity in childhood-onset schizophrenia to those found in adult chronic schizophrenia, thus supporting the hypothesis of continuity of the childhood and adult forms of the illness. Comparisons with data from other childhood disorders suggest that the combination of low-elicited SC activity with high levels of spontaneous SC activity may be specific to schizophrenia. PMID- 9337770 TI - Neurobiology and neuroplasticity in schizophrenia. Continuity across the life cycle. PMID- 9337771 TI - 'Vascular depression' hypothesis. AB - We propose that cerebrovascular disease may predispose, precipitate, or perpetuate some geriatric depressive syndromes. The "vascular depression" hypothesis is supported by the comorbidity of depression, vascular disease, and vascular risk factors and the association of ischemic lesions to distinctive behavioral symptoms. Disruption of prefrontal systems or their modulating pathways by single lesions or by an accumulation of lesions exceeding a threshold are hypothesized to be central mechanisms in vascular depression. The vascular depression concept can generate studies of clinical and heuristic value. Drugs used for the prevention and treatment of cerebrovascular disease may be shown to reduce the risk for vascular depression or improve its outcomes. The choice of antidepressants in vascular depression may depend on their effect on neurologic recovery from ischemic lesions. Research can clarify the pathways to vascular depression by focusing on the site of the lesion, the resultant brain dysfunction, the presentation of depression and time of onset, and the contribution of nonbiological factors. PMID- 9337772 TI - Manual-guided psychosocial treatment. A new virtual requirement for pharmacotherapy trials? AB - The conduct of randomized clinical trials to evaluate the efficacy of pharmacotherapies for mental disorders is guided by research standards (at a high level of rigor) that govern most design elements, including randomization of subjects, use of placebo controls, formulation and dosage of the therapeutic agent, and monitoring of serum levels. In contrast, no such widely accepted guidelines are recognized for standardization of an essential, if unacknowledged, element of all such studies: the concomitant provision of at least a minimal form of psychosocial treatment. Standardized provision of psychosocial treatments in pharmacotherapy trials will foster replicability of findings and address several common problems (e.g., attrition, medication noncompliance, reduction of error variance, and ethical issues associated with placebo controls). Careful selection and standardization of the psychosocial context in which medications are delivered will improve the validity, precision, and power of pharmacotherapy efficacy research, and should be considered a virtual requirement in research design. PMID- 9337773 TI - A psychotherapeutic context for clinical trials is promising, but manualization is not. PMID- 9337774 TI - Offspring of depressed parents. 10 Years later. AB - BACKGROUND: There have been numerous studies that have shown that offspring of depressed parents are at a high risk for major depressive disorder (MDD) and impairment. None have followed up the offspring into adulthood to obtain more precise estimates of risk. METHOD: One hundred eighty-two offspring from 91 families, in which 1 or more parents had MDD (high risk) or in which neither parent was depressed (low risk), were blindly reassessed in the third follow-up, using a structured diagnostic instrument 10 years after their initial identification. RESULTS: Compared with the offspring for whom neither parent was depressed, the offspring of depressed parents had increased rates of MDD, particularly before puberty, and phobias (both at approximately a 3-fold risk), panic disorder, alcohol dependence (at a 5-fold risk), and greater social impairment. The peak age at onset for MDD in both high- and low-risk offspring ranged from 15 to 20 years. The peak age at onset for anxiety disorder was considerably earlier, especially in female offspring in the high-risk group. The onset of alcohol dependence in the offspring in the high-risk group peaked in adolescence and then after the age of 25 years. The depressed offspring of depressed parents, compared with nondepressed parents, had more serious and impairing depressions during the follow-up period but were less likely to go for treatment. CONCLUSIONS: The offspring of depressed parents are a high-risk group for onset of anxiety disorder and MDD in childhood, MDD in adolescence, and alcohol dependence in adolescence and early adulthood. The findings support the potential value of early detection in the offspring of depressed parents. PMID- 9337775 TI - Reduction of synaptophysin immunoreactivity in the prefrontal cortex of subjects with schizophrenia. Regional and diagnostic specificity. AB - BACKGROUND: Multiple lines of evidence indicate that the prefrontal cortex is a site of dysfunction in schizophrenia. However, the apparent absence of gross structural abnormalities in this area suggests that the pathophysiological characteristics of schizophrenia may involve more subtle disturbances in prefrontal cortical circuitry, such as alterations in synaptic connectivity and transmission. In this study, immunoreactivity for synaptophysin, an integral membrane protein of small synaptic vesicles, was used to assess the integrity of cortical synaptic circuitry in schizophrenia. METHODS: Using immunocytochemical techniques and adjusted optical density measurements, we examined synaptophysin immunoreactivity in prefrontal cortical areas 9 and 46 and in area 17 (the primary visual cortex) from 10 pairs of case subjects with schizophrenia and control subjects matched on a pairwise basis for age, sex, race, and postmortem interval, and in 5 matched pairs of nonschizophrenic psychiatric case subjects and normal control subjects. RESULTS: Compared with levels found in matched control subjects, synaptophysin immunoreactivity in areas 46 and 9 was significantly decreased (P < .001 and P < .008, respectively) across all cortical layers in the case subjects with schizophrenia. In contrast, no differences were observed in area 17. In addition, levels of synaptophysin immunoreactivity in areas 46, 9, and 17 did not differ between 5 nonschizophrenic psychiatric case subjects and their matched controls, suggesting that decreased synaptophysin levels in the prefrontal cortex of patients with schizophrenia may be specific to that disorder. CONCLUSIONS: Additional studies are required to determine if the decrease in levels of synaptophysin immunoreactivity is caused by a decrease in the number or size of presynaptic terminals, a decrease in the number of synaptic vesicles per terminal, or a decrease in the expression of synaptophysin. However, all of these potential explanations are consistent with a disturbance in synaptic transmission in the prefrontal cortex of patients with schizophrenia. PMID- 9337776 TI - Delayed normalization of central D2 dopamine receptor availability after discontinuation of haloperidol decanoate. Preliminary findings. AB - BACKGROUND: Antipsychotic drugs in depot formulations may prevent psychotic relapses, even after complete withdrawal. To examine the duration of drug remaining in the brain, central D2 dopamine receptor occupancy was measured with positron emission tomography for a year after discontinuation of depot neuroleptic treatment. METHODS: Four schizophrenic patients were withdrawn from low-dose treatment with haloperidol decanoate (30-50 mg every 4 weeks). They were examined repeatedly with positron emission tomography and the radioligand carbon 11-labeled raclopride during the following year. At end point, a Scatchard analysis was performed to determine the density and affinity of D2 dopamine receptors. RESULTS: Occupancy of D2 dopamine receptors was highest 1 week after depot injection (66%, 77%, 82%, and 78% in 4 patients) and then decreased slowly. Six months after discontinuation of treatment, D2 dopamine receptor occupancy was 24%, 32%, and 34% in 3 patients. After 1 year, D2 dopamine receptor density and affinity in 2 patients were within the ranges of control subjects, suggesting no remaining haloperidol. CONCLUSIONS: Our preliminary finding of persistence of D2 dopamine receptor occupancy indicates that commonly used doses of haloperidol decanoate (200 mg every 4 weeks) maintain antipsychotic levels of receptor occupancy even 16 weeks after discontinuation of treatment. This may explain the lower relapse rates in patients withdrawn from depot neuroleptic treatment compared with those withdrawn from oral treatment. In addition, the remaining occupancy may confound the clinical evaluation of subsequent treatments. For controlled clinical trials of new antipsychotic drugs, we suggest a minimum washout of 6 months after the last depot injection. PMID- 9337777 TI - Measurement of glutamate and glutamine in the medial prefrontal cortex of never treated schizophrenic patients and healthy controls by proton magnetic resonance spectroscopy. AB - BACKGROUND: Positron emission tomographic and postmortem studies comparing schizophrenic patients with healthy control subjects have found medial prefrontal cortical and anterior cingulate abnormalities that suggest dysfunction in glutamatergic neurons. The glutamate used for nerve signal transduction is predominantly derived from glutamine. After signal transduction, glutamate released into the synapse is converted to glutamine in glial cells, transported back to the presynaptic neuron, and reconverted to glutamate for reuse. In this study, levels of glutamate and glutamine were examined by means of in vivo proton (1H) magnetic resonance spectroscopy. METHODS: Localized in vivo 1H spectra were acquired from a 4.5-cm3 volume in the left medial prefrontal cortex encompassing portions of Brodmann areas 24, 32, and 9 in 10 never-treated schizophrenic subjects and 10 healthy controls of comparable age, sex, handedness, education, and parental education. From each spectrum, metabolite levels were estimated for glutamate and glutamine, as well as 10 other metabolites and 3 macromolecules, by means of a noninteractive computer program that combined modeled in vitro spectra of every metabolite to reconstruct each in vivo spectrum. RESULTS: A significant increase in glutamine level was found in the medial prefrontal cortex of the schizophrenic patients compared with controls. N-acetylaspartate and other measured metabolites and macromolecules were not significantly changed in schizophrenics. CONCLUSION: Increased glutamine levels in the medial prefrontal region most likely reflect decreased glutamatergic activity in this region in never-treated schizophrenic patients compared with healthy controls. PMID- 9337778 TI - Treatment of bulimia nervosa with ondansetron. PMID- 9337779 TI - Patent laws: could changes enhance drug development? PMID- 9337780 TI - The identification and validation of distinct depressive syndromes in a population-based sample of female twins. PMID- 9337781 TI - D2 dopamine receptor occupancy: a crossover comparison of risperidone with clozapine therapy in schizophrenic patients. PMID- 9337782 TI - Sertraline and imipramine for the treatment of dysthymia. PMID- 9337801 TI - Seven 'deadly myths' about old age. PMID- 9337783 TI - The idiosyncratic definition of nicotine dependence. PMID- 9337803 TI - Type 2 diabetes: stepped-care approach to patient management. AB - An oral antidiabetic medication and/or insulin in addition to exercise and a nutritious meal plan form the basis for treatment of type 2 diabetes. Blood glucose self-monitoring helps guide therapeutic decisions. Therapy needs modification when glycated hemoglobin levels exceed 8%. Initially pharmacologic therapy may include any one of the following: a sulfonylurea, metformin, acarbose, troglitazone, or insulin. If monotherapy does not maintain near normoglycemia, combined oral antidiabetic medication or insulin may bring glucose levels into the therapeutic range. If combination therapy does not achieve target goals, then insulin given twice daily or more often becomes necessary. PMID- 9337802 TI - Geriatrics photo quiz. Diabetes mellitus. PMID- 9337804 TI - Opportunistic fungal infections: filamentous fungi and cryptococcosis. AB - Older patients with diabetes mellitus or pulmonary diseases and those receiving immunosuppressive drugs are at an increased risk of infection with environmentally-acquired, opportunistic fungal diseases. Aspergillus most often produces invasive pulmonary or sinus infection in severely immuno-compromised patients. Chronic necrotizing pulmonary and sino-orbital aspergillosis present subacutely and are often misdiagnosed. Mucormycosis classically presents with rhinocerebral disease in diabetic patients with ketoacidosis, whereas pulmonary infection mimics invasive pulmonary aspergillosis and occurs mostly in patients who are neutropenic. Cryptococcal meningitis in the older patient may manifest simply as confusion. Amphotericin B is the preferred initial treatment for all three fungal infections. PMID- 9337805 TI - Opportunistic fungal infections: superficial and systemic candidiasis. AB - Age alone is not usually sufficient for the development of disease due to Candida, but it appears to be associated with increased morbidity and mortality. Mucocutaneous Candida infections such as thrush and denture stomatitis are associated with local and mechanical factors. A rare and sight-threatening complication of cataract surgery is Candida endophthalmitis. Systemic Candida infections are becoming more common due to increasing use of immunosuppressive drugs and the increasing risk of nosocomial candidiasis in the intensive care unit. Candiduria is increasingly common in older patients with diabetes mellitus, indwelling urinary catheters, and a history of antibiotic therapy. PMID- 9337806 TI - Midlife periodic health exam in the primary care practice. AB - The periodic exam for the healthy midlife patient (age 45 to 65) includes blood pressure testing, cholesterol screening, and a baseline ECG. Counseling is appropriate for proper diet, exercise, and smoking cessation Screening tests are indicated for colon cancer for men and women, and for breast and cervical cancer for women. A check of immunization status can detect a need for recommended vaccines, including tetanus-diphtheria. Question patients about hearing problems, and test as needed. Counsel women about risk factors for osteoporosis and the need for adequate calcium and vitamin D intake. Ask about symptoms of urinary incontinence and sexual problems. Screen women for thyroid disease. PMID- 9337807 TI - How to get the most benefit from a changing home health care system. AB - By 1998, a Medicare prospective payment system for home care is expected to be in place. Physicians must become more involved in home care, because they will be held accountable for the patients they refer and the services they order. PMID- 9337808 TI - Scaly, crusted papules. These pruritic lesions can become epidemic in nursing home residents. PMID- 9337821 TI - Long-term oxygen therapy in moderate hypoxaemia. PMID- 9337822 TI - Thunderstorms: a risk factor for asthma attacks. PMID- 9337823 TI - Should patients with cystic fibrosis infected with Burkholderia cepacia undergo lung transplantation? PMID- 9337824 TI - Effect of long-term oxygen therapy on survival in patients with chronic obstructive pulmonary disease with moderate hypoxaemia. AB - BACKGROUND: To date only two controlled studies have been published on the effects of domiciliary oxygen treatment on survival in patients with chronic obstructive pulmonary disease (COPD) with advanced respiratory failure. The survival in such patients despite oxygen treatment remains poor. The prescription of long term oxygen therapy (LTOT) in less severe disease remains controversial. The aim of this study was to evaluate the rationale for prescribing oxygen to patients with COPD with moderate hypoxaemia. METHODS: One hundred and thirty five patients with COPD, with PaO2 7.4-8.7 kPa (56-65 mmHg) and advanced airflow limitation (mean (SD) forced expiratory volume in one second (FEV1) 0.83 (0.28) 1), were randomly allocated to a control (n = 67) and LTOT (n = 68) group. The patients were followed every three months for at least three years or until death. RESULTS: The cumulative survival rate was 88% at one year, 77% at two years, and 66% at three years. No significant differences were found in survival rates between patients treated with LTOT and controls, nor did longer oxygen use (over 15 hours per day) improve survival. Younger age, better spirometric values, and higher body mass index predicted better survival. CONCLUSIONS: Domiciliary oxygen treatment does not prolong survival in patients with COPD with moderate hypoxaemia. Airway limitation seems to determine survival in this group of patients. PMID- 9337825 TI - Effect of thunderstorms and airborne grass pollen on the incidence of acute asthma in England, 1990-94. AB - BACKGROUND: Thunderstorms and prior grass pollen counts were investigated as predictors of daily hospital admissions for asthma in England. This study was motivated by reports in the literature of spectacular asthma epidemics associated with thunderstorms, particularly in the grass pollen season. METHODS: Asthma admissions for two age groups (0-14 years and 15 and over) were measured using the Hospital Episodes System (HES) in the 14 regional health authorities (RHAs) in England. Thunderstorms were measured daily in each RHA using densities of sferics (lightning flashes). Relative asthma excesses for moderate positive and exceptionally high sferic densities, with or without previous high grass pollen counts, were measured using log linear autoregression--allowing for weekly, seasonal, and longer term background variation--and pooled over RHAs by calculating geometric means. RESULTS: Relative risks from all RHAs were pooled to form geometric means. Exceptional sferic densities were associated with a relative excess risk of around 25% in both age groups. Moderate sferic densities were associated with a smaller excess, statistically significant in the two age groups taken together. In five RHAs in which grass pollen counts were available, high pollen counts for the previous five days were associated with an amplification of the excess associated with thunderstorms. CONCLUSION: Very large sferic densities are associated with moderate rises in hospital admissions for acute asthma. However, typical thunderstorm days are not associated with spectacular asthma epidemics of the scale previously reported in the literature. Thunderstorm-associated excesses are amplified after a run of high pollen counts. PMID- 9337826 TI - Adrenocortical activity with repeated twice daily dosing of fluticasone propionate and budesonide given via a large volume spacer to asthmatic school children. AB - BACKGROUND: In a previous single dosing study in asthmatic school children fluticasone propionate produced significantly greater suppression of overnight urinary cortisol excretion than budesonide at high doses of 800 micrograms/day or greater. The aim of this study was to assess whether conventional lower doses of both drugs cause adrenal suppression when given at steady state twice daily by large volume spacer on a microgram equivalent basis in asthmatic school children. METHODS: Eight school children of mean age 12.1 years with stable asthma of mild to moderate severity (forced expiratory volume in one second (FEV1) 78.6% predicted, mid forced expiratory flow rate (FEF25-75) 72.5% predicted), on 400 micrograms/day or less of inhaled corticosteroid, were studied in a single blind (investigator blind), placebo controlled, crossover design comparing inhaled budesonide and fluticasone propionate 100 micrograms bid and 200 micrograms bid. Each dose was given at 08.00 hours and 20.00 hours for four days by metered dose inhaler via their respective large volume spacers with mouth rinsing. Measurements were made of overnight urinary cortisol and creatinine excretion after the eighth dose. RESULTS: Neither drug produced significant suppression of overnight urinary cortisol or cortisol/creatinine excretion compared with pooled placebo and there were no differences between the drugs. Only one subject with each drug at 200 micrograms twice daily had abnormally low urinary cortisol excretion of < 10 nmol/12 hours. Ratios for the fold difference between active treatment versus placebo for urinary cortisol excretion were (as means and 95% confidence intervals for difference): budesonide 100 micrograms b.i.d 1.03 (95% CI 0.46 to 1.61), budesonide 200 micrograms b.i.d 1.04 (95% CI 0.62 to 1.46); fluticasone 100 micrograms b.i.d 1.11 (0.45 to 1.77), fluticasone 200 micrograms b.i.d 1.12 (0.78 to 1.47). Likewise, there were no significant differences in overnight urinary cortisol/creatinine excretion. CONCLUSIONS: With repeated twice daily administration at steady state across a dose range of 200-400 micrograms/day no evidence of significant adrenal suppression was found using the sensitive marker of overnight urinary cortisol excretion for either fluticasone propionate or budesonide given via a large volume spacer. These results emphasise the good safety profile in children of these inhaled steroids at conventional dose levels, which have proven antiasthmatic efficacy. PMID- 9337827 TI - Physiological effects and optimisation of nasal assist-control ventilation for patients with chronic obstructive pulmonary disease in respiratory failure. AB - BACKGROUND: A study was undertaken to investigate the effects of non-invasive assist-control ventilation (ACV) by nasal mask on respiratory physiological parameters and comfort in acute on chronic respiratory failure (ACRF). METHODS: Fifteen patients with chronic obstructive pulmonary disease (COPD) were prospectively and randomly assigned to two non-invasive ventilation (NIV) sequences in spontaneous breathing (SB) and ACV mode. ACV settings were always optimised and therefore subsequently adjusted according to patient's tolerance and air leaks. RESULTS: ACV significantly decreased all the total inspiratory work of breathing (WOBinsp) parameters, pressure time product, and oesophageal pressure variation in comparison with SB mode. The ACV mode also resulted in a significant reduction in surface diaphragmatic electromyographic activity to 36% of the control values and significantly improved the breathing pattern. SB did not change the arterial blood gas tensions from baseline values whereas ACV significantly improved both the PaO2 from a mean (SD) of 8.45 (2.95) kPa to 13.31 (2.15) kPa, PaCO2 from 9.52 (1.61) kPa to 7.39 (1.39) kPa, and the pH from 7.32 (0.03) to 7.40 (0.07). The respiratory comfort was significantly lower with ACV than with SB. CONCLUSIONS: This study shows that the clinical benefit of non invasive ACV in the management of ACRF in patients with COPD results in a reduced inspiratory muscle activity providing an improvement in breathing pattern and gas exchange. Despite respiratory discomfort, the muscle rest provided appears sufficient when ACV settings are optimised. PMID- 9337828 TI - Effect of measurement conditions on measured levels of peak exhaled nitric oxide. AB - BACKGROUND: It is possible to measure nitric oxide (NO) levels in exhaled air. The absolute concentrations of exhaled NO obtained by separate workers in similar patient groups and normal subjects with apparently similar techniques have been very different. A study was undertaken to determine whether changes in measurement conditions alter the concentration of exhaled NO. METHOD: NO concentrations measured by a chemiluminescence analyser (Dasibi Environmental Corporation) and carbon dioxide (CO2) measured by a Morgan capnograph were analysed in single exhalations from total lung capacity in healthy volunteers (mean age 35.9 years). Ten subjects performed five exhalations at four different expiratory flow rates, at four different expiratory mouth pressures, and before and after drinking hot (n = 5) or cold (n = 5) water. Three subjects performed five exhalations on a day of high background NO (mean NO level 134 ppb) before and after a set of five exhalations made while both the subject and analysers were sampling from a low NO/NO-free reservoir system. RESULTS: The mean peak concentration of NO decreased by 35 ppb (95% CI 25.7 to 43.4) from a mean (SE) of 79.0 (15.5) ppb at an expiratory flow rate of 250 ml/min to 54.1 (10.7) ppb at 1100 ml/min. The mean peak concentration of NO did not change significantly with change in mouth pressure. The mean (SE) peak NO concentration decreased from 94.4 (20.8) ppb to 70.8 (16.5) ppb (p = 0.002, 95% CI 12.9 to 33.1) with water consumption. The mean NO concentration with machine and subject sampling from the low NO reservoir was 123.1 (19.4) ppb, an increase from results obtained before (81.9 (10.2) ppb, p = 0.001, 95% CI -19.9 to -62.7) and after (94.2 (18.3) ppb, p = 0.017, 95% CI 6.0 to 51.8) sampling with high ambient NO. CONCLUSIONS: The measurement of exhaled NO must be performed in a carefully standardised manner to enable different teams of investigators to compare results. PMID- 9337830 TI - An audit of bronchoscopy practice in the United Kingdom: a survey of adherence to national guidelines. AB - BACKGROUND: Both patient and staff safety are of major importance during the procedure of fibreoptic bronchoscopy. Patient safety depends partly on adequate disinfection of instruments and accessories used as well as careful monitoring during the procedure. Adequate facilities, manpower and training are also essential. Staff safety depends partly on adequate procedures to minimise any risks of sensitisation to agents such as glutaraldehyde. An audit was carried out of bronchoscopy procedures in hospitals in the UK and the findings were compared with published guidelines on good practice and clinical consensus. METHODS: A postal questionnaire was sent to 218 bronchoscopy units in the UK. Findings were then compared with published evidence of good practice in the areas of disinfection, including the use of glutaraldehyde, patient monitoring, manpower, facilities, and training. RESULTS: A 73% response rate was obtained. Recommended minimum disinfection times before and after routine bronchoscopies were not achieved by 35% of units. No disinfection was carried out in 34% of units before emergency bronchoscopies and in 19% of units after suspected cases of tuberculosis. Adequate rinsing of the bronchoscope with sterile or filtered water was not carried out by 43% of units. Contrary to recommendations, 31% of departments were still using glutaraldehyde in the patient examination room and inadequate room ventilation was common. Protective clothing was often not worn by staff during bronchoscopy. Inadequate intravenous access and use of supplementary oxygen were found in many units. Practice standards were higher in departments where dedicated bronchoscopy/endoscopy units of the hospital were used, and also where staff had been on external training courses. CONCLUSIONS: This audit has shown that many units do not adhere to guidelines on disinfection procedures and patient monitoring. Unnecessary potential risks due to staff exposure to glutaraldehyde were apparent. National guidelines on good practice are not being followed in areas which may potentially affect patient and staff safety. PMID- 9337829 TI - Optimal treatment of descending necrotising mediastinitis. AB - BACKGROUND: Descending necrotising mediastinitis is caused by downward spread of neck infection and has a high fatality rate of 31%. The seriousness of this infection is caused by the absence of barriers in the contiguous fascial planes of neck and mediastinum. METHODS: The recent successful treatment of seven adult patients with descending necrotising mediastinitis emphasises the importance of optimal early drainage of both neck and mediastinum and prolonged antibiotic therapy. The case is also presented of a child with descending necrotising mediastinitis, demonstrating the rapidity with which the infection can develop and lead to death. Twenty four case reports and 12 series of adult patients with descending necrotising mediastinitis published since 1970 were reviewed with meta analysis. In each case of confirmed descending necrotising mediastinitis the method of surgical drainage (cervical, mediastinal, or none) and the survival outcome (discharge home or death) were noted. The chi 2 test of statistical significance was used to detect a difference between cases treated with cervical drainage alone and cases where mediastinal drainage was added. RESULTS: Cervical drainage alone was often insufficient to control the infection with a fatality rate of 47% compared with 19% when mediastinal drainage was added (p < 0.05). CONCLUSIONS: Early combined drainage with neck and chest incisions, together with broad spectrum intravenous antibiotics, should be considered standard care for this disease. PMID- 9337831 TI - Experimental bilateral lobar lung transplantation and its application in humans. AB - BACKGROUND: The critical lack of donor organs from people of small size or children has created great difficulties in transplantation for recipients who are of smaller size. Surgical techniques of organ reduction and partial transplantation may to some extent solve the problem of disparity in organ size, be it liver or lung, and lessen the problem of scarcity of paediatric organs. METHODS: In a series of experiments on dogs the surgical technique of pulmonary partition of a large organ from a grown dog followed by transplantation of lobes, either unilaterally or bilaterally, into a young dog was studied. Two series of experiments were performed in two groups of animals; in group 1 transplantation of a single right lobe (n = 6) or single left lobe (n = 6) from a split adult lung was carried out and in group 2 (n = 10) animals received bilateral lobar transplants from a single split adult lung. The animals were sacrificed at fixed intervals (days 8-120 in group 1, days 7-10 in group 2) and the results of the surgical technique were assessed. RESULTS: Healing of lobar bronchial anastomoses was found to be excellent with no histological evidence of dehiscence or ulceration. There was one bronchial anastomotic stenosis and one arterial thrombosis. Morphological and functional adaptation of the lobes in the thorax was found to be excellent in both groups of animals. The technique has been applied in a clinical setting and the first patient with bilateral lobar lung transplantation followed for 30 months is reported. CONCLUSION: Lung partition and subsequent lobar transplantation, either unilaterally or bilaterally, is associated with satisfactory early results in an animal experimental model. Initial clinical experience in one patient has been successful. PMID- 9337832 TI - Respiratory mechanics after heart-lung and bilateral lung transplantation. AB - BACKGROUND: The factors determining respiratory mechanics following heart-lung transplantation (HLT) and bilateral lung transplantation (BLT) are incompletely understood. METHODS: The dynamic and static lung volumes of 15 patients after HLT (n = 6) and BLT (n = 9) with no evidence of obliterative bronchiolitis were analysed to assess the factors which determine lung volumes following transplantation. Post-transplantation total lung capacity (TLCpost) was compared with the size of the recipient's lungs (TLCpre), the predicted capacity of the thorax of the recipient (TLCpred), and the predicted size of the donor's lungs (TLCdon). In addition, the post-transplantation respiratory mechanics were investigated by measuring the static pressure-volume (PV) curve of the lungs and the maximum respiratory pressures in a subgroup of nine patients (four HLT, five BLT). RESULTS: TLCpost was closely related to TLCpred in both groups and showed no correlation with TLCpre. The mean (95% CI) TLCpost was 102.5 (90.2 to 115)% predicted for the recipient in the HLT group and 109 (97.6 to 120)% predicted for the recipient in the BLT group. Despite the near normal TLC, residual volume (RV) and functional residual capacity (FRC) remained increased after transplantation in both groups. These abnormalities were not attributable to either airflow obstruction or expiratory muscle weakness. On average, lung compliance expressed in terms of the shape constant of the static pressure-volume curve of the lungs was mildly reduced in both groups compared with values predicted for the recipient. CONCLUSIONS: These results suggest that at high lung volumes the chest wall adapts to the size of transplanted lungs, while at lower volumes the increase in FRC and RV might be due to a persistent change in the static pressure volume curve of the chest wall. PMID- 9337833 TI - Lung surfactant in a cystic fibrosis animal model: increased alveolar phospholipid pool size without altered composition and surface tension function in cftrm1HGU/m1HGU mice. AB - BACKGROUND: Progressive pulmonary dysfunction is a characteristic symptom of cystic fibrosis (CF) and is associated with functional impairment and biochemical alterations of surfactant phospholipids in the airways. However, the fundamental question of whether surfactant alterations in the CF lung are secondary to the pulmonary damage or are present before initiation of chronic infection and inflammation has yet to be resolved in patients with cystic fibrosis but can now be addressed in CF mice that exhibit the basic defect in the airways. A study was therefore undertaken to investigate the pool sizes, composition, and function of lung surfactant in the non-infected cftrm1HGU/m1HGU mouse. METHODS: The amount and composition of phospholipid classes and phosphatidylcholine molecular species were determined in bronchoalveolar lavage (BAL) fluid and lavaged lungs by high performance liquid chromatography (HPLC). Surfactant protein A (SP-A) levels in BAL fluid were determined by ELISA and surfactant for functional measurements was isolated from BAL fluid by differential ultracentrifugation. Equilibrium and minimal surface tension of surfactant was assessed by the pulsating bubble surfactometer technique. MF1, BALB/c, C57/BL6, and C3H/He mice served as controls. RESULTS: BAL fluid of cftrm1HGU/m1HGU mice contained 1.02 (95% confidence interval (CI) 0.89 to 1.16) mumol phospholipid and 259 (239 to 279) ng SP-A. BAL fluid of MF1, BALB/c, C57BL/6, and C3H/He mice contained 0.69 (0.63 to 0.75), 0.50 (0.42 to 0.57), 0.52 (0.40 to 0.64), and 0.45 (0.27 to 0.63) mumol phospholipid, respectively. After correction for the different body weights of mouse strains, phospholipid levels in BAL fluid of cftrm1HGU/m1HGU mice were increased by 64 (52 to 76)%, 60 (39 to 89)%, 72 (45 to 113)%, and 92 (49 to 163)%, respectively, compared with controls. The amount of SP-A in BAL fluid and the composition of phospholipid as well as phosphatidylcholine molecular species in BAL fluid and lung tissue was unchanged in cftrm1HGU/m1HGU mice compared with controls. The increase in phospholipids in BAL fluid of cftrm1HGU/m1HGU mice resulted from an increased fraction of large aggregates which exhibited normal surface tension function. CONCLUSION: In cftrm1HGU/m1HGU mice surfactant homeostasis is perturbed by an increased phospholipid pool in the alveolar compartment. PMID- 9337834 TI - Evaluation of diaphragmatic fatigue in obstructive sleep apnoeas during non-REM sleep. AB - BACKGROUND: Resistive load applied to the airways may induce diaphragmatic fatigue, and hypoxaemia has been shown to predispose to the development of fatigue. Inspiratory muscle fatigue may occur in patients with obstructive sleep apnoea syndrome (OSAS), as these patients repetitively develop both inspiratory loading and hypoxaemia. The results of previous studies on this topic are inconclusive, probably because of the methodological approaches used. METHODS: Six obese patients with OSAS underwent a polysomnographic study. The diaphragmatic pressure time index (PTI) was evaluated as an indicator of diaphragmatic contraction, and the mean frequency of the diaphragmatic electromyogram power spectrum (Fm) and the maximum relaxation rate of transdiaphragmatic pressure (MRR) as indices of a fatiguing diaphragm. A total of 119 randomly selected apnoeas (each including 5-13 occluded efforts) were analysed throughout the night in non-REM sleep to assess possible muscle fatigue due to the high pressure generation in each apnoea. A breath-by-breath within apnoea analysis was performed on the first three pre-apnoeic breaths, on all the occluded efforts, and on the first three unoccluded breaths following the apnoea interruption. Possible fatigue development due to the cumulative effect of apnoeas over the night was also evaluated. RESULTS: A progressive increase of Fm and MRR was found during the obstructive phase in all the subjects in the within apnoea analysis. The overnight analysis did not show a reduction in either PTI, Fm, or MRR secondary to recurrent upper airway obstruction during the night. CONCLUSIONS: No evidence of diaphragmatic fatigue or impaired diaphragmatic contraction was found either within each apnoea or throughout the whole night, despite the generation of high PTI values during the apnoeic occluded phases. It is concluded that diaphragmatic fatigue does not occur in OSAS during non-REM sleep. PMID- 9337835 TI - Exhaled NO during graded changes in inhaled oxygen in man. AB - BACKGROUND: Nitric oxide (NO) is present in the exhaled air of animals and humans. In isolated animal lungs the amount of exhaled NO is decreased during hypoxia. A study was undertaken to determine whether changes in arterial oxygen tension affect levels of exhaled NO in humans. METHODS: Sixteen healthy subjects were randomised to inhale different gas mixtures of oxygen and nitrogen in a double blind crossover study. Eight gas mixtures of oxygen and nitrogen (fractional inspired oxygen concentration (FiO2) 0.1 to 1.0) were administered. Exhaled NO was measured with a chemiluminescence detector from end expiratory single breath exhalation. RESULTS: A dose-dependent change in exhaled NO during graded oxygen breathing was observed (p = 0.0012). The mean (SE) exhaled NO concentration was 31 (3) ppb at baseline, 39 (4) ppb at an FiO2 of 1.0, and 26 (3) ppb at an FiO2 of 0.1. CONCLUSIONS: The NO concentration in exhaled air in healthy humans is dependent on oxygen tension. Hyperoxia increases the level of exhaled NO, which indicates increased NO production. The mechanism behind this phenomenon remains to be elucidated. PMID- 9337836 TI - Sample size estimation in studies monitoring exercise-induced bronchoconstriction in asthmatic children. AB - BACKGROUND: The repeatability of the response to standardised treadmill exercise testing using dry air and monitoring of heart rate in asthmatic children suffering from exercise-induced bronchoconstriction (EIB) has not been well established. METHODS: Twenty seven asthmatic children with known EIB performed standardised exercise testing twice within a period of three weeks. The tests were performed on a treadmill while breathing dry air. During both tests heart rate had to reach 90% of the predicted maximum. Response to exercise was expressed as % fall in forced expiratory volume in one second (FEV1) from baseline and as area under the curve (AUC) of the time-response curve. RESULTS: The intra-class correlation coefficients for % fall and AUC (log-transformed) were 0.57 and 0.67, respectively. From these data, power curves were constructed that allowed estimations to be made of sample sizes required for studies of EIB in children. These indicated that, if a drug is expected to reduce EIB by 50%, as few as 12 patients would be sufficient to demonstrate this effect (90% power) using a parallel design study. CONCLUSIONS: Standardised exercise testing for EIB using dry air and monitoring of heart rate is adequately repeatable for use in research and clinical practice in children with asthma. PMID- 9337838 TI - Listeria monocytogenes empyema in an HIV infected patient. AB - Listeriosis in HIV infected patients is uncommon and usually presents as meningitis or bacteraemia. Pleural fluid infections caused by this organism are extremely rare. A case is described of empyema caused by Listeria monocytogenes in an HIV infected patient that was successfully treated with medical treatment only. PMID- 9337837 TI - Gene therapy for lung inflammatory diseases: not so far away? AB - The lung is a readily accessible target organ for gene therapy. To date, therapeutic gene delivery has largely focused on introducing functional, corrective genes in lung diseases arising from single gene defects such as cystic fibrosis. More recently interest has centred on gene therapy as a potential therapeutic tool in modulating complex pathological processes such as pulmonary inflammation. Genetic modification of critical components of the inflammatory process may be beneficial-for example, overexpressing anti-elastase genes may circumvent elastase mediated lung damage in emphysema. With the development of improved viral and liposome vectors and the evolution of effective adjuvant immunosuppression to obviate host immune responses--for example, using selective cytokines and blockers of T cell surface activation--the potential exists to target therapeutic doses of transgene to deficient or dysregulated cells. Furthermore, increased understanding of tissue-specific promoter regions and of mechanisms controlling regulation of gene expression offer the potential for close control of therapeutic gene expression within the lung. Continuing refinements in these technologies will provide new therapeutic strategies in inflammatory lung disease. PMID- 9337839 TI - Occupational asthma in an isothiazolinone manufacturing plant. AB - A chemical plant operator developed asthma five months after starting work in an isothiazolinone manufacturing plant. He described symptoms of late asthmatic reactions after work with isothiazolinone. Airway responsiveness to methacholine improved tenfold when he was removed from the plant for 18 days. A workplace challenge study then resulted in a deterioration in airway responsiveness to its earlier level and in progressive falls in forced expiratory volume in one second (FEV1) over three days at work compared with control days, indicating statistically significant late asthmatic reactions of increasing severity. PMID- 9337840 TI - Regression of polyvinylchloride polymer pneumoconiosis. AB - A 35 year old man heavily exposed to polyvinylchloride (PVC) polymer dust developed dyspnoea and a mild restrictive lung disorder consistent with PVC pneumoconiosis. Clinical and radiological abnormalities cleared on removal from exposure, suggesting that in its early stages PVC pneumoconiosis is reversible. PMID- 9337841 TI - Fenoterol, asthma deaths, and asthma severity: confounding or confusion? PMID- 9337842 TI - Thermogenic effect of bronchodilators in COPD. PMID- 9337843 TI - Rare presentation or lead time bias in pleural sarcoidosis? PMID- 9337845 TI - Rac1 is required for cell proliferation and G2/M progression. AB - We have transiently expressed a dominant negative form of rac1 (N17rac1) using adenoviral-mediated gene transfer. The level of N17rac1 expression is demonstrated to be proportional to the multiplicity of infection. Expression of N17rac1 in Rat 2 fibroblasts results in cytostatic growth arrest. Cell-cycle analysis demonstrates that cells expressing N17rac1 accumulate in G2/M. These results suggest that rac1 is required for cell proliferation and provide the first demonstration in mammalian cells of a role for small GTP-binding proteins in the G2/M transition. PMID- 9337844 TI - Caspases: the executioners of apoptosis. AB - Apoptosis is a major form of cell death, characterized initially by a series of stereotypic morphological changes. In the nematode Caenorhabditis elegans, the gene ced-3 encodes a protein required for developmental cell death. Since the recognition that CED-3 has sequence identity with the mammalian cysteine protease interleukin-1 beta-converting enzyme (ICE), a family of at least 10 related cysteine proteases has been identified. These proteins are characterized by almost absolute specificity for aspartic acid in the P1 position. All the caspases (ICE-like proteases) contain a conserved QACXG (where X is R, Q or G) pentapeptide active-site motif. Capases are synthesized as inactive proenzymes comprising an N-terminal peptide (prodomain) together with one large and one small subunit. The crystal structures of both caspase-1 and caspase-3 show that the active enzyme is a heterotetramer, containing two small and two large subunits. Activation of caspases during apoptosis results in the cleavage of critical cellular substrates, including poly(ADP-ribose) polymerase and lamins, so precipitating the dramatic morphological changes of apoptosis. Apoptosis induced by CD95 (Fas/APO-1) and tumour necrosis factor activates caspase-8 (MACH/FLICE/Mch5), which contains an N-terminus with FADD (Fas-associating protein with death domain)-like death effector domains, so providing a direct link between cell death receptors and the caspases. The importance of caspase prodomains in the regulation of apoptosis is further highlighted by the recognition of adapter molecules, such as RAIDD [receptor-interacting protein (RIP)-associated ICH-1/CED-3-homologous protein with a death domain]/CRADD (caspase and RIP adapter with death domain), which binds to the prodomain of caspase-2 and recruits it to the signalling complex. Cells undergoing apoptosis following triggering of death receptors execute the death programme by activating a hierarchy of caspases, with caspase-8 and possibly caspase-10 being at or near the apex of this apoptotic cascade. PMID- 9337846 TI - The third member of the Tetrahymena CCT subunit gene family, TpCCT alpha, encodes a component of the hetero-oligomeric chaperonin complex. AB - The sequence of a third member of the Tetrahymena pyriformis chaperonin CCT ('chaperonin containing TCP1') subunit gene family is presented. This gene, designated TpCCT alpha, is the orthologue of the mouse chaperonin gene TCP1/CCT alpha. To characterize the CCT complex in this ciliate, we have produced polyclonal antibodies against synthetic peptides based on C-terminal sequences deduced from the primary sequences of the TpCCT alpha, TpCCT gamma and TpCCT eta subunits. We have also used polyclonal antibodies produced against recombinant yeast CCT alpha and CCT beta subunits. Using these antibodies, we show that Tetrahymena cells contain a hetero-oligomeric CCT chaperonin comprising at least seven distinct subunits. Three of these were assigned to specific TpCCT genes, whereas a fourth was recognized by the polyclonal antibody against yeast CCT beta, suggesting that this gene is also present in the ciliate. The CCT complex also contains other unidentified proteins that were recognized by the polyclonal antibody UM-1, raised against the putative ATP binding domain of the chaperonin proteins. TpCCT alpha gene expression was shown in exponentially growing cells and cells regenerating their cilia for different periods to have a similar pattern to the previously identified genes TpCCT gamma and TpCCT eta, and also to tubulin genes. PMID- 9337847 TI - The effects of truncations of the small subunit on m-calpain activity and heterodimer formation. AB - In order to study subunit interactions in calpain, the effects of small subunit truncations on m-calpain activity and heterodimer formation have been measured. It has been shown previously that active calpain is formed by co-expression of the large subunit (80 kDa) of rat m-calpain with a delta 86 form (21 kDa) of the small subunit. cDNA for the full-length 270 amino acid (28.5 kDa) rat calpain small subunit has now been cloned, both with and without an N-terminal histidine tag (NHis10). The full-length small subunit constructs yielded active calpains on co-expression with the large subunit, and the small subunit was autolysed to 20 kDa on exposure of these calpains to Ca2+. A series of deletion mutants of the small subunit, NHis10-delta 86, -delta 99, -delta 107, and -delta 116, gave active heterodimeric calpains with unchanged specific activities, although in decreasing yield, and with a progressive decrease in stability. NHis10-delta 125 formed a heterodimer which was inactive and unstable. Removal of 25 C-terminal residues from delta 86, leaving residues 87-245, abolished both activity and heterodimer formation. The results show that: (a) generation of active m-calpain in Escherichia coli requires heterodimer formation; (b) small subunit residues between 94 and 116 contribute to the stability of the active heterodimer but do not directly affect the catalytic mechanism; (c) residues in the region 245-270 are essential for subunit binding. Finally, it was shown that an inactive mutant Cys103-->Ser-80k/delta 86 calpain, used in order to preclude autolysis, did not dissociate in the presence of Ca2+, a result which does not support the proposal that Ca(2+)-induced dissociation is involved in calpain activation. PMID- 9337848 TI - Synthetic histatin analogues with broad-spectrum antimicrobial activity. AB - Histatins are salivary histidine-rich cationic peptides, ranging from 7 to 38 amino acid residues in length, that exert a potent killing effect in vitro on Candida albicans. Starting from the C-terminal fungicidal domain of histatin 5 (residues 11-24, called dh-5) a number of substitution analogues were chemically synthesized to study the effect of amphipathicity of the peptide in helix conformation on candidacidal activity. Single substitutions in dh-5 at several positions did not have any effect on fungicidal activity. However, multi-site substituted analogues (dhvar1 and dhvar2) exhibited a 6-fold increased activity over dh-5. In addition, dhvar1 and dhvar2 inhibited the growth of the second most common yeast found in clinical isolates, Torulopsis glabrata, of oral- and non oral pathogens such as Prevotella intermedia and Streptococcus mutans, and of a methicillin-resistant Staphylococcus aureus. In their broad-spectrum activity, dhvar1 and dhvar2 were comparable to magainins (PGLa and magainin 2), antimicrobial peptides of amphibian origin. Both the fungicidal and the haemolytic activities of dhvar1, dhvar2 and magainins increased at decreasing ionic strength. PMID- 9337849 TI - Identification by site-directed mutagenesis of three essential histidine residues in membrane dipeptidase, a novel mammalian zinc peptidase. AB - Membrane dipeptidase (EC 3.4.13.19) is a plasma membrane zinc peptidase that is involved in the renal metabolism of glutathione and its conjugates, such as leukotriene D4. The enzyme lacks the classical signatures of other zinc-dependent hydrolases and shows no homology with any other mammalian protein. We have used site-directed mutagenesis to explore the roles of five histidine residues in pig membrane dipeptidase that are conserved among mammalian species. When expressed in COS-1 cells, the mutants H49K and H128L exhibited a specific activity and Km for the substrate Gly-D-Phe comparable with those of the wild-type enzyme. However, the mutants H20L, H152L and H198K were inactive, but were expressed at the cell surface at equivalent levels to the wild-type, as assessed by immunoblotting and immunofluorescence. These three mutants were compared with regard to their ability to bind to the competitive inhibitor cilastatin, which binds with equal efficacy to native and EDTA-treated pig kidney membrane dipeptidase. Expressed wild-type enzyme and mutants H20L and H198K were efficiently bound by cilastatin-Sepharose, but H152L failed to bind. Thus His-152 appears to be involved in the binding of substrate or inhibitor, whereas His-20 and His-198 appear to be involved in catalysis. Membrane dipeptidase shares some similarity with a dipeptidase recently cloned from Acinetobacter calcoaceticus. In particular, His-20 and His-198 of membrane dipeptidase are conserved in the bacterial enzyme, as are Glu-125 and His-219, previously shown to be required for catalytic activity. PMID- 9337850 TI - Human phosphodiesterase 4A: characterization of full-length and truncated enzymes expressed in COS cells. AB - The type 4 phosphodiesterase (PDE) family comprises four enzymes (4A, 4B, 4C and 4D) that are characterized by their specificity for cAMP and selective inhibition by the anti-depressant drug rolipram (4-[3-(cyclopentoxyl)-4-methoxyphenyl]2 pyrrolidone). In common with other PDEs, they consist of a central conserved domain associated with catalytic activity in addition to two N-terminal upstream conserved regions (UCR1 and UCR2) that are unique to the type 4 enzymes. We have isolated a 2 kb cDNA encoding a full-length type 4A PDE{HSPDE4A4B[Bolger, Michaeli, Martins, St.John, Steiner, Rodgers, Riggs, Wigler and Ferguson (1993) Mol. Cell. Biol. 13, 6558-6571]} from a human frontal cortex cDNA library. Northern blot analysis showed that the major PDE4A mRNA of 4.5 kb was widely distributed in different human tissues. The recombinant PDE4A expressed in COS cells had a molecular mass of approx. 117 kDa as revealed by SDS/PAGE/Western blotting with a PDE4A-specific antibody and was specific for cAMP with a Km of 4.8 microM. The enzyme activity was potently inhibited by R-rolipram (IC50 204 nM) and showed a 2.7-fold stereoselectivity over the S enantiomer. Analysis of the kinetics of inhibition indicated that R-rolipram did not behave as a simple competitive inhibitor. Dixon replots suggested that there was more than one mode of interaction consistent with the detection in the enzyme of a high-affinity binding site for R-rolipram with a Kd of 2.3 nM. Truncation of the PDE4A enzyme by deletion mutagenesis showed that neither of the UCRs was required for catalytic activity and identified an approx. 71 kDa core enzyme with a K(m) for cAMP of 3.3 microM. In contrast with the full-length PDE4A, R-rolipram behaved as a simple competitive inhibitor of this form of the enzyme with decreased potency (IC50 1022 nM) and no stereoselectivity. In addition, no high-affinity rolipram binding site was detected in the truncated enzyme, indicating that this interaction involves sequences upstream of the catalytic domain of the enzyme. PMID- 9337851 TI - Sustained activation of extracellular-signal-regulated kinase 1 (ERK1) is required for the continued expression of cyclin D1 in G1 phase. AB - In Chinese hamster embryo fibroblasts (IIC9 cells), platelet-derived growth factor (PDGF) stimulated mitogen-activated protein kinase/extracellular-signal regulated kinase (MAP kinase/ERK) activity, but not that of c-jun N-terminal kinase (JNK), and induced G1 phase progression. ERK1 activation was biphasic and was sustained throughout the G1 phase of the cell cycle. PDGF induced cyclin D1 protein and mRNA levels in a time-dependent manner. Inhibition of PDGF-induced ERK1 activity by the addition of a selective inhibitor of MEK1 (MAP kinase kinase/ERK kinase 1) activation, PD98059, or transfection with a dominant negative ERK1 (dnERK-) was correlated with growth arrest. In contrast, growth was unaffected by expression of dominant-negative JNK (dnJNK-). Interestingly, addition of PD98059 or dnERK-, but not dnJNK-, resulted in a dramatic decrease in cyclin D1 protein and mRNA levels, concomitant with a decrease in cyclin D1 cyclin-dependent kinase activity. To investigate the importance of sustained ERK1 activation, ERK1 activity was blocked by the addition of PD98059 throughout G1. Addition of PD98059 up to 4 h after PDGF treatment decreased ERK1 activity to the levels found in growth-arrested IIC9 cells. Loss of cyclin D1 mRNA and protein expression was observed within 1 h after inhibition of the second sustained phase of ERK1 activity. Disruption of sustained ERK1 activity also resulted in G1 growth arrest. These data provide evidence for a role for sustained ERK activity in controlling G1 progression through positive regulation of the continued expression of cyclin D1, a protein known to positively regulate G1 progression. PMID- 9337852 TI - Distribution of mRNA for human epiregulin, a differentially expressed member of the epidermal growth factor family. AB - We have recently identified epiregulin as a new growth regulator and a member of the epidermal growth factor (EGF) family. Epiregulin has certain characteristics that are different from those of the classical members of the EGF family, EGF and transforming growth factor alpha, including mitogenic responses on several normal cells and binding to EGF receptors on epidermoid carcinoma A431 cells. In the present study we cloned and identified the expression of human epiregulin transcript. The human epiregulin gene encoded a 163-residue putative transmembrane precursor containing an EGF-like domain in the internal segment, and the structural organization was similar to that of other members of the EGF family that bind to EGF receptors. Northern blot analysis showed the expression of human epiregulin to be mainly on peripheral blood macrophages and the placenta in normal tissues, and was highest on epithelial tumour cell lines in various types of tumour cell lines. The expression profile was quite different from that of other members of the EGF family in normal and tumour cells. Recombinant expression in mammalian cells also showed that human epiregulin was secreted as a soluble form of approx. 5 kDa that is biologically active on the basis of the stimulation of DNA synthesis. Our findings suggest that epiregulin is involved in certain physiological processes such as maintenance or development of normal cell growth, and the progression of carcinomas. PMID- 9337853 TI - Receptor recognition sites reside in both lobes of human serum transferrin. AB - The binding of iron by transferrin leads to a significant conformational change in each lobe of the protein. Numerous studies have shown that the transferrin receptor discriminates between iron-saturated and iron-free transferrin and that it modulates the release of iron. Given these observations, it seems likely that there is contact between each lobe of transferrin and the receptor. This is the case with chicken transferrin, in which it has been demonstrated unambiguously that both lobes are required for binding and iron donation to occur [Brown-Mason and Woodworth (1984) J. Biol. Chem. 259, 1866-1873]. Further support to this contention is added by the ability of both N- and C-domain-specific monoclonal antibodies to block the binding of a solution containing both lobes [Mason, Brown and Church (1987) J. Biol. Chem. 262, 9011-9015]. In the present study a similar conclusion is reached for the binding of human serum transferrin to the transferrin receptor. With the use of recombinant N- and C-lobes of human transferrin produced in a mammalian expression system, we show that both lobes are required to achieve full binding. (Production of recombinant C-lobe in the baby hamster kidney cell system is reported here for the first time.) Each lobe is able to donate iron to transferrin receptors on HeLa S3 cells in the presence of the contralateral lobe. The results are not identical with the chicken system, because the C-lobe alone shows a limited ability to bind to receptors and to donate iron. Further complications arise from the relatively weak re-association between the two lobes of human transferrin compared with the re-association of the ovotransferrin lobes. However, domain-specific monoclonal antibodies to either lobe block the binding of N- and C-lobe mixtures in the human system, thus substantiating the need for both. PMID- 9337854 TI - Characterization of the oxidation products of the reaction between reduced glutathione and hypochlorous acid. AB - Reduced glutathione (GSH) is one of the most preferred biological substrates of myeloperoxidase-derived hypochlorous acid and is a likely target for neutrophil oxidants. We have used HPLC to show that the oxidation of GSH by hypochlorous acid gives two major, stable products in addition to glutathione disulphide (GSSG). The most prevalent product lacks free amine and thiol groups, and was shown by electrospray MS to have a molecular mass of 337 Da. This corresponds to GSH with a gain of two oxygen atoms and a loss of two hydrogen atoms, and is consistent with the product being an internal sulphonamide. The other novel product has a molecular mass of 644 Da, and has amine groups but no free thiols. These properties are consistent with it being glutathione thiolsulphonate. Whereas GSSG in the cell is recycled enzymically, formation of these higher oxidation products is likely to be irreversible. Hypochlorous acid, therefore, could compromise the cell by depleting GSH. The putative sulphonamide may be unique for oxidation by hypochlorous acid and thus provide a useful marker of neutrophil oxidant activity. PMID- 9337855 TI - Involvement of proteasomal subunits zeta and iota in RNA degradation. AB - We have identified two distinct subunits of 20 S proteasomes that are associated with RNase activity. Proteasome subunits zeta and iota, eluted from two dimensional Western blots, hydrolysed tobacco mosaic virus RNA, whereas none of the other subunits degraded this substrate under the same conditions. Additionally, proteasomes were dissociated by 6 M urea, and subunit zeta, containing the highest RNase activity, was isolated by anion-exchange chromatography and gel filtration. Purified subunit zeta migrated as a single spot on two-dimensional PAGE with a molecular mass of approx. 28 kDa. Addition of anti-(subunit zeta) antibodies led to the co-precipitation of this proteasome subunit and nuclease activity. This is the first evidence that proteasomal alpha type subunits are associated with an enzymic activity, and our results provide further evidence that proteasomes may be involved in cellular RNA metabolism. PMID- 9337856 TI - Molecular characterization of gp40, a mucin-type glycoprotein from the apical plasma membrane of Madin-Darby canine kidney cells (type I). AB - gp40 has been recently identified as a major apical cell-surface sialoglycoprotein of type-I Madin-Darby canine kidney cells, a cell line widely used for the study of polarized transport. The determination of two internal amino acid sequences of the purified glycoprotein by Edman degradation enabled us to isolated the cDNA encoding the 18.6 kDa protein backbone of gp40. Sequence analysis revealed that gp40 is a type-I membrane protein which has several characteristics in common with glycophorin A and other mucin-type glycoproteins. At least 14 serine/threonine residues were found to be used for O-glycosylation. No potential sites for N-glycosylation were detected. gp40 turned out to represent the canine homologue of a cell-surface antigen expressed by various epithelial and non-epithelial cells in rat and mouse. Potential O-glycosylation sites, transmembrane and cytoplasmic domains were found to be highly conserved in the three species. gp40 was detected in canine lung, intestine, kidney, brain and heart but not in liver and spleen. The subline II of Madin-Darby canine kidney cells was found not to express gp40. Stable expression of gp40 in transfected type-II cells revealed that gp40 is predominantly delivered to the apical plasma membrane. N-Glycans and a glycosylphosphatidylinositol anchor, both proposed apical targeting signals, are absent from gp40, indicating that other determinants are responsible for its polarized transport. PMID- 9337857 TI - Reaction of variant sperm-whale myoglobins with hydrogen peroxide: the effects of mutating a histidine residue in the haem distal pocket. AB - The reaction of hydrogen peroxide with a number of variants of sperm-whale myoglobin in which the distal pocket histidine residue (His64) had been mutated was studied with a combination of stopped-flow spectroscopy and freeze-quench EPR. The rate of the initial bimolecular reaction with hydrogen peroxide in all the proteins studied was found to depend on the polarity of the amino acid side chain at position 64. In wild-type myoglobin there were no significant optical changes subsequent to this reaction, suggesting the rapid formation of the well characterized oxyferryl species. This conclusion was supported by freeze-quench EPR data, which were consistent with the pattern of reactivity previously reported [King and Winfield (1963) J. Biol. Chem. 238, 1520-1528]. In those myoglobins bearing a mutation at position 64, the initial bimolecular reaction with hydrogen peroxide yielded an intermediate species that subsequently decayed via a second hydrogen peroxide-dependent step leading to modification or destruction of the haem. In the mutant His64-->Gln the calculated electronic absorption spectrum of the intermediate was not that of an oxyferryl species but seemed to be that of a low-spin ferric haem. Freeze-quench EPR studies of this mutant and the apolar mutant (His64-->Val) revealed the accumulation of a novel intermediate after the first hydrogen peroxide-dependent reaction. The unusual EPR characteristics of this species are provisionally assigned to a low-spin ferric haem with bound peroxide as the distal ligand. These results are interpreted in terms of a reaction scheme in which the polarity of the distal pocket governs the rate of binding of hydrogen peroxide to the haem iron and the residue at position 64 governs both the rate of heterolytic oxygen scission and the stability of the oxyferryl product. PMID- 9337858 TI - Hyperoxia, unlike phorbol ester, induces glutathione peroxidase through a protein kinase C-independent mechanism. AB - Human selenium-dependent glutathione peroxidase (GP) is implicated as a mechanism of resistance against oxygen free radicals. The 5' flanking sequence upstream from the coding region of GP contained an oxygen-responsive element termed ORE1 that is responsive to hypoxia, as well as several copies of the activator protein 1 (AP-1)- and AP-1-like-binding sites. In this study, we sought to define the molecular events that lead to GP gene transcription in response to hyperoxia in human umbilical-vein endothelial cells, and asked whether such induction is mimicked and sustained by activation of protein kinase C (PKC) by phorbol esters. Treatment of cells with 100 nM phorbol 12,13-dibutyrate (PdBu) induced a delayed (24-48 h) but significant (2-fold) increase in steady-state GP mRNA levels. Steady-state GP mRNA levels also rose after exposure to 95% O2, again after considerable delay (48-72 h). For both PdBu and oxygen, induction was transcriptionally regulated, as demonstrated by nuclear run-on experiments. The simulations by PdBu and oxygen were additive. In contrast with PdBu, hyperoxia did not stimulate translocation of PKC from the cytosol to the particulate fraction, although the specific activity of both cytosolic and particulate associated PKC was increased 2-fold in cells exposed to 95% O2 for 5 days. In addition, gel mobility-shift assays using double-stranded tumour-promoting-agent responsive element (TRE) and nuclear extracts derived from phorbol- and oxygen treated cells revealed that PdBu, but not hyperoxia, increased AP-1 DNA-binding activity. On the other hand, the up-regulation of GP expression by oxygen could not be accounted for by the ORE1 core sequence, since no specific protein-DNA binding activity could be detected using nuclear extracts from hyperoxic cells and ORE1. Taken together, these results suggest that there may be different molecular mechanisms controlling GP expression. After exposure to PdBu, GP undergoes transcriptional activation via a process that can be readily explained by a classic AP-1 interaction with the TRE sites in the GP promoter. During hyperoxia, GP also undergoes transcriptional activity, but via a process that appears to involve neither TRE nor ORE1. PMID- 9337859 TI - Immortalization and characterization of a cell line exhibiting a severe multiple sulphatase deficiency phenotype. AB - Multiple sulphatase deficiency (MSD) is a rare genetic defect that causes a simultaneous deficiency of all known sulphatases. All available evidence suggests that the deficient gene product is normally responsible for the post translational modification of a conserved cysteine residue to 2-amino-3 oxopropionic acid and that this modification is necessary for sulphatase activity. MSD often has an enzymically mild phenotype, with significant levels of residual sulphatase activity being detectable. Here we identify an MSD cell line in which the residual activity of the sulphatases assayed was generally very low. To characterize the phenotype of this cell line further, immortalized lines were established after transformation with simian virus 40 (SV40) T antigen. Immortalized cell lines representing normal and MSD phenotypes were then transduced with a retroviral vector carrying the gene encoding human N acetylgalactosamine-4-sulphatase. Analysis of N-acetylgalactosamine-4-sulphatase protein synthesis and enzyme activity showed that transduced cell lines expressed large amounts of enzyme and that the specific activity of this enzyme was approx. 0.5-1.5% of normal, confirming that this cell line defines a severe phenotype for MSD. N-Acetylgalactosamine-4-sulphatase purified from a transduced MSD cell line seemed normal on denaturing PAGE. Kinetic analysis of the purified enzyme suggests that the residual activity is due to small amounts of normal enzyme rather than unmodified enzyme with low levels of residual activity. These cell lines and the availability of large amounts of inactive N-acetylgalactosamine-4 sulphatase from MSD cells should facilitate the further study of this disorder. PMID- 9337860 TI - Purification and staining of intact yeast DNA chromosomes and real-time observation of their migration during gel electrophoresis. AB - In the past few years, fluorescence microscopy has been used successfully to characterize the motion of intermediate-size DNA molecules (50-500 kbp) during steady- and pulsed-field gel electrophoresis. However, experimental difficulties had prevented the application of this technique to the direct observation of longer DNA chromosomes (1-2 Mbp). In the present study a particular procedure was followed for the purification and staining of chromosomal yeast DNA to protect it from shear forces. Also, a new highly fluorescent DNA-labelling dye, YOYO-1, was employed to improve brightness and contrast. Finally, the motion of such long DNA molecules (1-2 Mbp) was characterized under steady-field electrophoresis conditions. An accurate description of the molecular mechanisms of motion of such long molecules should provide the basis for a detailed analysis of the mechanisms responsible for DNA trapping. PMID- 9337861 TI - Cloning of a human phosphoinositide 3-kinase with a C2 domain that displays reduced sensitivity to the inhibitor wortmannin. AB - The generation of phosphatidylinositide 3-phosphates has been observed in a variety of cellular responses. The enzymes that mediate synthesis are the phosphoinositide 3-kinases (PI3-Ks) that form a family of structurally diverse enzymes with distinct substrate specificities. In this paper, we describe the cloning of a novel human PI3-K, namely PI3-K-C2 alpha, which contains a C terminal C2 domain. This enzyme can be assigned to the class II PI3-Ks, which was defined by characterization of the Drosophila 68D enzyme and includes the recently described murine enzymes m-cpk and p170. Despite the overall similarity in the amino acid sequence of the murine and human enzymes, which suggests that they are encoded by closely related genes, these molecules show marked sequence heterogeneity at their N-termini. Biochemical analysis of recombinant PI3-K-C2 alpha demonstrates a restricted lipid substrate specificity. As reported for other members of this class, the enzyme only phosphorylates PtdIns and PtdIns4P when the lipids are presented alone. However, when lipids were presented together with phosphatidylserine acting as a carrier, phosphorylation of PtdIns(4,5)P2 was also observed. The catalytic activity of PI3-K-C2 alpha is refractory to concentrations of wortmannin and LY294002 which inhibit the PI3-K activity of other family members. The comparative insensitivity of PI3-K-C2 alpha to these inhibitors suggests that their use should be reevaluated in the study of PI3-Ks. PMID- 9337862 TI - Phosphorylation of serine-167 on the human oestrogen receptor is important for oestrogen response element binding and transcriptional activation. AB - We have studied the role of phosphorylation of the human oestrogen receptor (hOR; otherwise known as hER) at serine-167, which has been identified previously as the major oestrogen-induced phosphorylation site. We have tested transactivation by the hOR in yeast and cell-free transcription assays, and shown that mutation of serine-167 results in a 70% decrease in hOR-dependent transcription. Furthermore we explored the functional significance of phosphorylation at this site by hormone binding and DNA binding. DNA binding affinity was 10-fold lower when serine-167 was changed to alanine in the hOR. Cell-free transcription experiments showed that casein kinase II is the enzyme responsible for oestradiol dependent phosphorylation of the hOR at serine-167. This suggests that a conformational change of the hOR must occur upon hormone binding that exposes serine-167 to casein kinase II, resulting in transactivation of oestrogen responsive genes. PMID- 9337863 TI - Cyanate-mediated inhibition of neutrophil myeloperoxidase activity. AB - Cyanate (CNO-) forms spontaneously in solutions containing urea, and is present in urine and the body fluids of uraemic patients. We have explored the possibility that CNO- might be one of the unknown substances responsible for the reported impairment, by urine and uraemic plasma, of neutrophil oxidative metabolism (especially as measured by luminol-enhanced chemiluminescence). Luminol-enhanced chemiluminescence generated by human neutrophils derives predominantly from the activity of myeloperoxidase (MPO) which produces hypochlorous acid from H2O2 and Cl-. We hypothesized that CNO- (which resembles the 'pseudohalide' thiocyanate, an alternative substrate for MPO) might somehow interfere with the activity of MPO. In support of this, we find: (i) CNO- inhibits both peroxidative and halogenating activities of MPO and also inhibits the enzyme within intact human neutrophils; (ii) the inhibition is H2O2 dependent, irreversible, accompanied by covalent addition of [14C]CNO- (or a carbon-containing fragment thereof) to the enzyme; (iii) CNO- also inhibits Cl /H2O2/MPO-mediated bacterial killing. Impairment of this arm of neutrophil bactericidal activity by CNO- formed from urea may be one factor in the risk of urinary-tract infection associated with urinary stasis and perhaps in the generalized increase in susceptibility to infection in uraemic patients. PMID- 9337865 TI - Interaction of nitric oxide with non-haem iron sites of Escherichia coli bacterioferritin: reduction of nitric oxide to nitrous oxide and oxidation of iron(II) to iron(III). AB - The bacterioferritin (BFR) of Escherichia coli consists of 24 identical subunits, each containing a dinuclear metal-binding site consisting of two histidines and four carboxylic acid residues. Earlier studies showed that the characterization of iron binding to BFR could be aided by EPR analysis of iron-nitrosyl species resulting from the addition of NO to the protein [Le Brun, Cheesman, Andrews, Harrison, Guest, Moore and Thomson (1993) FEBS Lett. 323, 261-266]. We now report data from gas chromatographic head space analysis combined with EPR spectroscopy to show that NO is not an inert probe: iron(II)-BFR catalyses the reduction of NO to N2O, resulting in oxidation of iron(II) at the dinuclear centre and the subsequent detection of mononuclear iron(III). In the presence of excess reductant (sodium ascorbate), iron(II)-BFR also catalyses the reduction of NO to N2O, giving rise to three mononuclear iron-nitrosyl species which are detectable by EPR. One of these, a dinitrosyl-iron complex of S = 1/2, present at a maximum of one per subunit, is shown by EPR studies of site-directed variants of BFR not to be located at the dinuclear centre. This is consistent with a proposal that the diferric form of the centre is unstable and breaks down to form mononuclear iron species. PMID- 9337864 TI - Differential regulation of gamma-glutamylcysteine synthetase heavy and light subunit gene expression. AB - gamma-Glutamylcysteine synthetase (GCS) is the rate-limiting enzyme in the biosynthesis of glutathione and is composed of a heavy and a light subunit. Although the heavy subunit is enzymically active alone, the light subunit plays an important regulatory role by making the holoenzyme function more efficiently. In the current study we examined whether conditions which are known to influence gene expression of the heavy subunit also influence that of the light subunit, and the mechanisms involved. Treatment of cultured rat hepatocytes with hormones such as insulin and hydrocortisone, or plating hepatocytes under low cell density increased the steady-state mRNA level of the heavy subunit only. Treatment with diethyl maleate (DEM), buthionine sulphoximine (BSO) and t-butylhydroquinone (TBH) increased the steady state mRNA level and gene transcription rates of both subunits. These treatments share in common their ability to induce oxidative stress and activate nuclear factor kappa B (NF-kappa B). Treatment with protease inhibitors 7-amino-1-chloro-3-tosylamido-2-heptanone (TLCK) or L-1-tosylamido-2 phenylethyl chloromethyl ketone (TPCK) had no influence on the basal NF-kappa B and GCS subunit mRNA levels, but blocked the activation of NF-kappa B by DEM, BSO and TBH, and the increase in GCS heavy subunit mRNA level by BSO and TBH. On the other hand, the DEM-, BSO- and TBH-induced increase in GCS light-subunit mRNA level was unaffected by TLCK and TPCK. Thus only the heavy subunit is hormonally regulated and growth sensitive, whereas both subunits are regulated by oxidative stress. Signalling through NF-kappa B is involved only in the oxidative-stress mediated changes in the heavy subunit gene expression. PMID- 9337866 TI - Conformational state of a 25-mer peptide from the cyclophilin-binding loop of the HIV type 1 capsid protein. AB - Recently a 25-residue part of Gag polyprotein from HIV type 1 (HIV-1) was reported to bind to the cytosolic 18 kDa cyclophilin (Cyp18) with an IC50 value of 180 microM. This peptide corresponds to the Cyp18-binding domain of HIV-1 Gag. A replacement of Gly with Ala in the cyclophilin-binding loop of HIV-1 Gag polyprotein results in the prevention of the packaging of Cyp18 into virions. We found only two conformers of this peptide among 16 possible expected conformers, owing to cis/trans isomerization of four peptidyl-prolyl bonds. Although this finding implicates the existence of a stabilizing structure, we were not able to detect secondary structure formation by 1H-NMR and CD spectroscopy. We characterized the peptide as a substrate for Cyp18 by two-dimensional exchange 1H NMR spectroscopy. Surprisingly, we found similar binding characteristics for a peptide corresponding to 25-mer peptide containing the above-mentioned Gly to Ala substitution. PMID- 9337867 TI - Inhibition of cytokine-induced inducible nitric oxide synthase expression by glucagon and cAMP in cultured hepatocytes. AB - Addition of lipopolysaccharide plus interferon gamma, tumour necrosis factor alpha and interleukin 1 beta to cultured hepatocytes resulted in the induction of inducible nitric oxide synthase (iNOS) activity as measured by NO3(-)+NO2- formation, the conversion of L-[U-14C]arginine into citrulline and Western blotting of the iNOS protein. The inclusion of 1 microM glucagon during the induction period significantly decreased the effect of the cytokines on iNOS activity, the major effect being at the level of the total amount of protein, rather than alterations in substrate supply or covalent modification of the existing protein. In contrast, 1 microM insulin was without effect. The effect of glucagon was mediated via cAMP and could be mimicked by the presence of either dibutyryl cAMP or forskolin to activate adenylate cyclase directly. It was rapid in onset and long-lived, a 30 min pretreatment period protecting the cells from the induction of NO synthesis over the next 21 h in the presence of cytokines. Addition of glucagon at any time point up to 9 h after treatment of the cells with lipopolysaccharide plus the cytokines resulted in a significant inhibition of iNOS activity, glucagon being most potent when added during the first 3 h. PMID- 9337868 TI - Binding of glutathione and an inhibitor to microsomal glutathione transferase. AB - Microsomal glutathione transferase is an abundant liver protein that can be activated by thiol reagents. It is not known whether the activation is associated with changed binding properties of the enzyme. Therefore the binding of GSH and an inhibitor to rat liver microsomal glutathione transferase was studied by use of equilibrium dialysis and equilibrium partition in a two-phase system. The radioactive substrate glutathione and an inhibitor (glutathione sulphonate) give hyperbolic binding isotherms with a stoichiometry of 1 mol per mol of enzyme (i.e. 1 molecule per homotrimer). Glutathione had an equilibrium binding constant of 18 microM. Competition experiments involving glutathione sulphonate showed that it could effectively displace GSH. These and kinetic studies showed that the Kd and Ki for glutathione sulphonic acid are close to 10 microM. No change in these parameters was obtained after N-ethylmaleimide activation of the enzyme. Thus activation does not result from changes in binding affinity to GSH. PMID- 9337869 TI - Studies on recombinant Acetobacter xylinum alpha-phosphoglucomutase. AB - The phosphoglucomutase (PGM) from Acetobacter xylinum, which had been cloned and expressed in Escherichia coli, has been studied. After expression, the enzyme was purified from the E. coli in a three-step process consisting of (NH4)2SO4 precipitation, gel filtration and anion-exchange chromatography. The purified enzyme gave one band on gel electrophoresis and was judged essentially free of impurities, although it was unstable when diluted without the addition of 15 microM BSA. The isoelectric point for A. xylinum PGM was 4.8 and the molar absorbance was 3.9 x 10(4) M-1.cm-1. The enzyme was reasonably heat-stable below 50 degrees C and was stable throughout the pH 5.5-7.4 range, but was 70% inactivated at pH 10.0 and completely inactivated after standing for 10 min at pH 3.0 or at pH 12.4. When isolated, the recombinant enzyme was fully active without the addition of extra Mg2+. The Km for glucose 1-phosphate was much higher than that of other PGM species reported, which accords with the production of extracellular cellulose in A. xylinum. Glucose 1,6-diphosphate is not considered to be a substrate or coenzyme but an activating cofactor like Mg2+. The following kinetic constants were determined: Vmax 81.1 units/mg; kcat and the turnover rate 135 s-1; Km (glucose 1,6-diphosphate) 0.2 microM; Km (glucose 1-phosphate) 2.6 mM; kcat/Km (glucose 1-phosphate) 5.2 x 10(4) M-1.s-1. The recombinant enzyme is considered to follow a characteristic substituted enzyme or Ping Pong reaction mechanism. PMID- 9337870 TI - Altered regulation of cholesterol and cholesteryl ester synthesis in Chinese hamster ovary cells overexpressing the oxysterol-binding protein is dependent on the pleckstrin homology domain. AB - Oxysterol-binding protein (OSBP) is a high-affinity receptor for a variety of oxysterols, such as 25-hydroxycholesterol, that down-regulate cholesterol synthesis and stimulate cholesterol esterification. To examine a potential role for OSBP in regulating cholesterol metabolism, we stably overexpressed this protein in Chinese-hamster ovary (CHO)-K1 cells. Compared with mock-transfected controls, several cell lines overexpressing wild-type OSBP (CHO-OSBP) displayed a 50% decrease in cholesteryl ester synthesis when cultured in medium with delipidated serum, 25-hydroxycholesterol or low-density lipoprotein (LDL). CHO OSBP cells showed a 40-60% decrease in acyl-CoA:cholesterol acyltransferase activity and mRNA, a 50% elevation in mRNA for three sterol-regulated genes [LDL receptor, 3-hydroxy-3-methylgluraryl (HMG)-CoA reductase and HMG-CoA synthase], and an 80% increase in [14C]acetate incorporation into cholesterol. CHO-K1 cells overexpressing two OSBP mutants with a complete or N-terminal deletion of the pleckstrin homology (PH) domain had cholesterol esterification and synthesis rates that were similar to those shown by mock-transfected controls. Unlike wild type OSBP, both PH domain mutants displayed diffuse cytoplasmic immunofluorescence staining and did not translocate to the Golgi apparatus in the presence of 25-hydroxycholesterol. CHO-K1 cells overexpressing OSBP have pronounced alterations in cholesterol esterification and synthesis, indicating a potential role for this receptor in cholesterol homoeostasis. The phenotype observed in cells overexpressing OSBP is dependent on the PH domain, which appears to be necessary for ligand-dependent localization of OSBP to the Golgi apparatus. PMID- 9337871 TI - Incremental Ca2+ mobilization by inositol trisphosphate receptors is unlikely to be mediated by their desensitization or regulation by luminal or cytosolic Ca2+. AB - The kinetics of Ins(1,4,5)P3 (InsP3)-stimulated Ca2+ release from intracellular stores are unusual in that submaximal concentrations of InsP3 rapidly release only a fraction of the InsP3-sensitive Ca2+ stores. By measuring unidirectional 45Ca2+ efflux from permeabilized rat hepatocytes, we demonstrate that such quantal responses to InsP3 occur at all temperatures between 2 and 37 degrees C, but at much lower rates at the lower temperatures. Preincubation with submaximal concentrations of InsP3, which themselves evoked quantal Ca2+ release, had no effect on the sensitivity of the stores to further additions of InsP3. The final Ca2+ content of the stores was the same whether they were stimulated with two submaximal doses of InsP3 or a single addition of the sum of these doses. Such incremental responses and the persistence of quantal behaviour at 2 degrees C indicate that InsP3-evoked receptor inactivation is unlikely to be the cause of quantal Ca2+ mobilization. Reducing the Ca2+ content of the intracellular stores by up to 45% did not affect their sensitivity to InsP3, but substantially reduced the time taken for each submaximal InsP3 concentration to exert its full effect. These results suggest that neither luminal nor cytosolic Ca2+ regulation of InsP3 receptors are the determinants of quantal behaviour. Our results are not therefore consistent with incremental responses to InsP3 depending on mechanisms involving attenuation of InsP3 receptor function by cytosolic or luminal Ca2+ or by InsP3 binding itself. We conclude that incremental activation of Ca2+ release results from all-or-nothing emptying of stores with heterogeneous sensitivities to InsP3. These characteristics allow rapid graded recruitment of InsP3-sensitive Ca2+ stores as the cytosolic InsP3 concentration increases. PMID- 9337872 TI - Structural identification of the myo-inositol 1,4,5-trisphosphate-binding domain in rat brain inositol 1,4,5-trisphosphate 3-kinase. AB - A series of key amino acids involved in Ins(1,4,5)P3 (InsP3) binding and catalytic activity of rat brain InsP3 3-kinase has been identified. The catalytic domain is at the C-terminal end and restricted to a maximum of 275 amino acids [Takazawa and Erneux (1991) Biochem. J. 280, 125-129]. In this study, newly prepared 5'-deletion and site-directed mutants have been compared both for InsP3 binding and InsP3 3-kinase activity. When the protein was expressed from L259 to R459, the activity was lost but InsP3 binding was conserved. Another deletion mutant that had lost only four amino acids after L259 had lost InsP3 binding, and this finding suggests that these residues (i.e. L259DCK262) are involved in InsP3 binding. To further support the data, we have produced two mutants by site directed mutagenesis on residues C261 and K262. The two new enzymes were designated M4 (C261S) and M5 (K262A). M4 showed similar Vmax and Km values for InsP3 and ATP to wild-type enzyme. In contrast, M5 was totally inactive but had kept the ability to bind to calmodulin-Sepharose. C-terminal deletion mutants that had lost five, seven or nine amino acids showed a large decrease in InsP3 binding and InsP3 3-kinase activity. One mutant that had lost five amino acids (M2) was purified to apparent homogeneity: Km values for both substrates appeared unchanged but Vmax was decreased approx. 40-fold compared with the wild-type enzyme. The results indicate that (1) a positively charged amino acid residue K262 is essential for InsP3 binding and (2) amino acids at the C-terminal end of the protein are necessary to act as a catalyst in the InsP3 3-kinase reaction. PMID- 9337873 TI - Modulation of human type II secretory phospholipase A2 by sphingomyelin and annexin VI. AB - Conjectural results have been reported on the capacity of inflammatory secreted phospholipase A2 (sPLA2) to hydrolyse mammalian membrane phospholipids. Development of an assay based on the release of non-esterified fatty acids by the enzyme acting on the organized phospholipid mixture constituting the membrane matrix has led to the identification of two prominent effectors, sphingomyelin (SPH) and annexin. Recombinant human type II sPLA2 hydrolyses red-cell membrane phospholipids with a marked preference for the inner leaflet. This preference is apparently related to the high content of SPH in the outer leaflet, which inhibits sPLA2. This inhibition by SPH is specific for sPLA2. Cholesterol counteracts the inhibition of sPLA2 by SPH, suggesting that the SPH-to cholesterol ratio accounts in vivo for the variable susceptibility of cell membranes to sPLA2. Different effects were observed of the presence of the non hydrolysable D-alpha-dipalmitoyl phosphatidylcholine (D-DPPC), which renders the membranes rigid but does not inhibit sPLA2. Annexin VI was shown, along with other annexins, to inhibit sPLA2 activity by sequestering the phospholipid substrate. The present study has provided the first evidence that annexin VI, in concentrations that inhibit hydrolysis of purified phospholipid substrates, stimulated the hydrolysis of membrane phospholipids by sPLA2. The activation requires the presence of membrane proteins. The effect is specific for type II sPLA2 and is not reproducible with type I PLA2. The activation by annexin VI of sPLA2 acting on red cell membranes results in the preferential release of polyunsaturated fatty acids. It suggests that type II sPLA2, in conjunction with annexin VI, might be involved in the final step of endocytosis and/or exocytosis providing the free polyunsaturated fatty acids acting synergistically to cause membrane fusion. PMID- 9337874 TI - Aggrecan degradation in human intervertebral disc and articular cartilage. AB - Aggrecan degradation in human intervertebral disc and articular cartilage has been studied by using anti-neoepitope antibodies specific for the N-terminal degradation products generated by cleavage within the interglobular domain at the metalloproteinase and aggrecanase sites. Immunoblot analysis of extracts of annulus fibrosus, nucleus pulposus and articular cartilage demonstrated age related patterns in the abundance of both degradation products. In all three tissues the metalloproteinase-generated fragment was present at very low levels in young individuals but increased in abundance with age. In the disc tissues, the abundance of this degradation product levelled off in the juvenile; for cartilage this occurred in early adulthood. Despite these temporal differences, the levels attained in adults were comparable for the three tissues. In contrast, the aggrecanase-generated degradation product exhibited tissue-specific differences in the variation of its abundance with age. Whereas this degradation product increased with age in annulus fibrosus and articular cartilage and had levelled off by adulthood, in nucleus pulposus it was present in greatest abundance in young individuals and decreased to very low levels with age. Examination of discs exhibiting various degrees of degeneration did not reveal any differences in the levels of the metalloproteinase and aggrecanase-generated cleavage products that could not be accounted for by differences in age. In adults the product of aggrecanase action was much more abundant in articular cartilage than in either of the disc tissues, despite the age-related increase also observed for annulus fibrosus. Analysis of tissue extracts with an antibody recognizing the G1 domain of aggrecan identified two major degradation products whose abundance and size were correlated with the fragments detected by the anti neoepitope antibodies. Taken together, these results indicate that cleavage at the metalloproteinase and aggrecanase sites are quantitatively important events in aggrecan catabolism in both articular cartilage and intervertebral disc in vivo. Moreover the two enzyme systems act independently and exhibit differences in the degree to which they contribute to aggrecan degradation in these tissues. PMID- 9337875 TI - Characterization of iduronate sulphatase mutants affecting N-glycosylation sites and the cysteine-84 residue. AB - Iduronate sulphatase (IDS) is responsible for mucopolysaccharidosis type II, a rare recessive X-linked lysosomal storage disease. The aim of this work was to evaluate the functional importance of each N-glycosylation site, and of the cysteine-84 residue. IDS mutant cDNAs, lacking one of the eight potential N glycosylation sites, were expressed in COS cells. Although each of the potential sites was used, none of the eight glycosylation sites appeared to be essential for lysosomal targeting. Another important sulphatase co- or post-translational modification for generating catalytic activity involves the conversion of a cysteine residue surrounded by a conserved sequence C-X-P-S-R into a 2-amino-3 oxopropionic acid residue [Schmidt, Selmer, Ingendoh and von Figura (1995) Cell 82, 271-278]. This conserved cysteine, located at amino acid position 84 in IDS, was replaced either by an alanine (C84A) or by a threonine (C84T) using site directed mutagenesis. C84A and C84T mutant cDNAs were expressed either in COS cells or in human lymphoblastoid cells deleted for the IDS gene. C84A had a drastic effect both for IDS processing and for catalytic activity. The C84T mutation produced a small amount of mature forms but also abolished enzyme activity, confirming that the cysteine residue at position 84 is required for IDS activity. PMID- 9337877 TI - Endogenous basic fibroblast growth factor isoforms involved in different intracellular protein complexes. AB - Four forms of basic fibroblast growth factor (bFGF or FGF-2) result from an alternative initiation of translation involving one AUG (155-amino acid form) and three CUGs (210-, 201- and 196-amino acid forms). These different forms of bFGF show different intracellular biological activities. To identify their intracellular targets, the 210- and 155-amino acid forms of bFGF were independently transfected into CHO cells and their correct subcellular localizations were verified, the 155-amino acid bFGF form being essentially cytoplasmic whereas the 210-amino acid protein was nuclear. The radiation fragmentation method was used to determine the target size of the different bFGF isoforms in the transfected CHO cells and to show that the 210- and 155-amino acids bFGF isoforms were included in protein complexes of 320 and 130 kDa respectively. Similar results were obtained using the SK-Hep1 cell line, which naturally expressed all forms of bFGF. Co-immunoprecipitation assays using different chimaeric bFGF-chloramphenicol acetyltransferase proteins showed that different cellular proteins are associated with different parts of the bFGF molecule. We conclude that bFGF isoforms are involved in different molecular complexes in the cytosol and nucleus, which would reflect different functions for these proteins. PMID- 9337876 TI - Evidence that a kinase distinct from protein kinase C and phosphatidylinositol 3 kinase mediates ligation-dependent serine/threonine phosphorylation of the T lymphocyte co-stimulatory molecule CD28. AB - The CD28 cytoplasmic tail contains several potential phosphorylation sites for the serine/threonine kinase protein kinase C (PKC) and/or proline-directed serine/threonine kinases, such as extracellular signal-regulated kinases. We demonstrate that ligation of CD28 by B7.1 results in strong serine/threonine phosphorylation of CD28. It is unlikely that ligation-stimulated phosphorylation of CD28 is mediated via activation of PKC, since it was not prevented by pre treatment of Jurkat cells with inhibitors of PKC, and it was not mimicked by treatment with PKC activators such as PMA. Nevertheless, despite for lack of detectable effects of PMA treatment on CD28 phosphorylation, PMA did partially inhibit the association of CD28 with the putative signalling molecule phosphatidylinositol 3-kinase (PI 3-kinase) and the subsequent accumulation of PtdIns(3,4,5)P3. PI 3-kinase exhibits dual specificity as both a lipid kinase and a protein serine kinase, and site-specific mutagenesis of the Tyr173 residue in the CD28 cytoplasmic tail, which abolishes CD28 coupling to PI 3-kinase [Pages, Ragueneau, Rottapel, Truneh, Nunes, Imbert and Olive (1994) Nature (London) 369, 327-329], also prevents ligation-stimulated phosphorylation of CD28. However, the two PI 3-kinase inhibitors wortmannin and LY294002 had no effect on phosphorylation of CD28 after ligation by B7.1. This study therefore demonstrates that (1) a CD28-activated serine/threonine kinase distinct from both PKC and PI 3 kinase mediates ligation-stimulated CD28 phosphorylation, and (2) the PMA stimulated down-regulation of the coupling of CD28 to PI 3-kinase is not due to PMA-stimulated phosphorylation of CD28. PMID- 9337878 TI - Cysteine residues in human lysosomal acid lipase are involved in selective cholesteryl esterase activity. AB - Human lysosomal acid lipase (LAL) catalyses the deacylation of triacylglycerol and cholesteryl esters in the acidic lysosomal compartment. Treatment of LAL with the reducing agent dithiothreitol affected the triacylglycerol and cholesteryl esterase activities differentially, suggesting the involvement of cysteine residues in determining substrate specificity. To identify the residues involved, human LAL cDNA, under the control of the T7 promoter and tagged with a herpes simplex virus coding epitope, was specifically mutated in order to introduce single amino acid substitutions of each of the six cysteine residues of mature LAL. All Cys-227 mutants showed selectively decreased activity towards cholesteryl oleate, while preserving that towards trioleylglycerol. Substitutions of Cys-236, Cys-240 and Cys-244 affected catalysis towards the two substrates to a variable degree, depending on the side chain of the amino acid introduced. The replacement of Cys-41 or Cys-188 did not result in the preferential cleavage of either one of the two substrates. These data indicate that Cys-227, Cys-236, Cys 240 and Cys-244 play a crucial role in determining LAL substrate specificity. We propose that these cysteine residues are involved in the hydrolysis of cholesteryl ester by affecting selectively the access of this substrate to the catalytic active site. In addition, the fact that the catalytic activity is never completely abolished in cysteine mutants demonstrates that LAL is not a thiol enzyme. PMID- 9337879 TI - Phosphotyrosine phosphatase activity associated with c-Src in large multimeric complexes isolated from adrenal medullary chromaffin cells. AB - Chromaffin cells, which secrete catecholamines in response to acetylcholine, express high levels of the Src-family tyrosine kinases. These kinases contain protein-protein interaction domains which bind signal transduction proteins that participate in a variety of cellular processes. To determine if signalling proteins bind c-Src in chromaffin cells, we examined c-Src immunocomplexes for co precipitating proteins. We discovered a phosphotyrosine phosphatase (PTPase; EC 3.1.3.48) activity which associates with specific subcellular pools of c-Src in vivo and which preferentially binds the SH2 (Src homology 2) domain of c-Src in vitro. Known PTPases were not identified by blotting of c-Src immunocomplexes with a panel of anti-PTPase antibodies, suggesting that the PTPase may be a novel family member. The c-Src-PTPase complex is enriched in the plasma membrane fraction and exists in several large complexes, as revealed by gel-filtration analysis. This PTPase activity is altered rapidly following stimulation by secretagogues, decreasing within 30 s and returning to basal levels by 60 s of stimulation. Both the subcellular localization and rapid activity changes suggest that the c-Src-associated PTPase may function in early signalling events emanating from the nicotinic acetylcholine receptor. In support of this is the co precipitation of a PTPase activity with the nicotinic acetylcholine receptor and co-chromatography of this receptor with one or the c-Src-PTPase complexes. PMID- 9337880 TI - Characterization of the hydrolytic activity of a polyclonal catalytic antibody preparation by pH-dependence and chemical modification studies: evidence for the involvement of Tyr and Arg side chains as hydrogen-bond donors. AB - The hydrolyses of 4-nitrophenyl 4'-(3-aza-2-oxoheptyl)phenyl carbonate and of a new, more soluble, substrate, 4-nitrophenyl 4'-(3-aza-7-hydroxy-2 oxoheptyl)phenyl carbonate, each catalysed by a polyclonal antibody preparation elicited in a sheep by use of an analogous phosphate immunogen, were shown to adhere closely to the Michaelis-Menten equation, in accordance with the growing awareness that polyclonal catalytic antibodies may be much less heterogeneous than had been supposed. The particular value of studies on polyclonal catalytic antibodies is discussed briefly. Both the kcat and kcat/K(m) values were shown to increase with increase in pH across a pKa of approx. 9. Group-selective chemical modification studies established that the side chains of tyrosine and arginine residues are essential for catalytic activity, and provided no evidence for the involvement of side chains of lysine, histidine or cysteine residues. The combination of evidence from the kinetic and chemical modification studies and from studies on the pH-dependence of binding suggests that catalysis involves assistance to the reaction of the substrate with hydroxide ions by hydrogen-bond donation at the reaction centre by tyrosine and arginine side chains. This combination of hydrogen-bond donors appears to be a feature common to a number of other hydrolytic catalytic antibodies. High-pKa acidic side chains may be essential for the effectiveness of catalytic antibodies that utilize hydroxide ions. PMID- 9337881 TI - myo-Inositol is an osmolyte in rat liver macrophages (Kupffer cells) but not in RAW 264.7 mouse macrophages. AB - The role of myo-inositol as an osmolyte was studied in cultured rat liver macrophages (Kupffer cells). Hyperosmotic exposure of Kupffer cells stimulated myo-inositol uptake and led to an increase in the mRNA levels for the sodium/myo inositol cotransporter (SMIT). Conversely, hypo-osmotic (205 m-osM) exposure diminished myo-inositol uptake when compared with normo-osmotic (305 m-osM) control incubations. The hyperosmolarity-induced SMIT mRNA increase was counteracted by added myo-inositol or betaine. In contrast with Kupffer cells, there was only a slight hyperosmotic stimulation of myo-inositol uptake in RAW 264.7 mouse macrophages, and the myo-inositol transporter (SMIT) mRNA was not detectable. Further, a slight stimulation of taurine uptake and an increase in taurine transporter (TAUT) mRNA level by hyperosmolarity was observed in RAW 264.7 cells, whereas hypo-osmolarity led to a decrease in taurine uptake and TAUT mRNA level. When Kupffer cells were preloaded with myo-inositol, hypo-osmotic exposure led to a rapid efflux of myo-inositol from the cells. Myo-inositol efflux was also stimulated by phagocytosis of latex particles; however, latex was without effect on the hyperosmolarity-induced increase of SMIT mRNA levels. The results suggest a role of myo-inositol as an osmolyte in rat Kupffer cells but not in RAW 264.7 mouse macrophages. The functional relevance of this osmolyte strategy might lie in the maintenance of cell volume homeostasis during phagocytosis in Kupffer cells; however, the interplay with the other osmolytes betaine and taurine remains to be established. PMID- 9337882 TI - Suicidal ideation, bereavement, HIV serostatus and psychosocial variables in partners of men with AIDS. AB - This prospective 2-year study examines suicidal ideation in 86 HIV-positive and 167 HIV-negative caregiving partners of men with AIDS. One hundred and fifty-six of the caregivers became bereaved during the course of the study. The study focuses on the relationship between suicidal ideation and bereavement status (bereaved vs non-bereaved), HIV serostatus (HIV-positive vs HIV-negative), and psycho-social factors (caregiving burdens, social support, coping, and optimism). Bereavement was related to suicidal ideation, but HIV serostatus was not. High suicidal ideators were characterized by feeling burdened by caregiving, perceiving low levels of social support and subjective social integration, and the use of behavioural escape-avoidance coping. Those who reported never having suicidal ideation were characterized by higher levels of optimism. Clinical implications are discussed. PMID- 9337883 TI - Suicidality among HIV-positive psychiatric in-patients. AB - Rates of suicidality with HIV-infected, seriously mentally ill individuals were investigated. Fifty asymptomatic HIV-positive psychiatric in-patients were compared to a demographically-matched HIV-negative cohort. The groups were similar, except that seropositive subjects were less likely to be diagnosed with schizophrenia. Both groups had high rates of suicidality, with higher rates associated with non-schizophrenic diagnoses. HIV-positive subjects had higher rates of suicidality, with those diagnosed with schizophrenia showing the greatest difference from their HIV-negative counterparts. HIV-positive patients required less in-patient treatment. These data expand previous reports showing an association between HIV and increased suicidality, even among individuals with already elevated suicidal rates. PMID- 9337884 TI - Perceptions of children and community members concerning the circumstances of orphans in rural Zimbabwe. AB - Focus group discussions and interviews were held with 40 orphans, 25 caretakers and 33 other community workers from a rural area near Mutare, Zimbabwe. Orphan concerns included feeling different from other children, stress, stigmatization, exploitation, schooling, lack of visits and neglect of support responsibilities by relatives. Many community members, while recognizing their limitations due to poverty, were already actively helping orphans and caretakers. Extended family networks are the primary resource for orphans, though some relatives exploit orphans or fail to fulfil their responsibilities. Interventions are suggested which support community coping mechanisms by strengthening the capacities of families to care for orphans. Outside organizations can develop partnerships with community groups, helping them to respond to the impact of AIDS, by building upon existing concern for orphan families. They can help affected communities to develop orphan support activities which encourage caring responses by community leaders and relatives and which discourage property-grabbing and orphan neglect. Material support channelled through community groups to destitute families at critical times can strengthen family coping mechanisms. Income-generating activities should build upon communities' existing capabilities and benefit the most vulnerable orphan households. Some communities are responding to the AIDS disaster by adaptations to cope with devastating changes taking place in their communities. PMID- 9337885 TI - Sexual relationships, condom use and risk perception among female bar workers in north-west Tanzania. AB - This paper discusses the background characteristics, sexual relationships, condom use and risk perception of bar workers in Magu district, north-west Tanzania. Bar workers in Magu are geographically mobile. They are not highly educated but probably have a higher level of schooling than average. They are mostly unmarried or divorced. They choose bar work because, given the resources available, it provides them with a good balance between earning their own income and being independent. Although the women are still partially dependent on the financial support provided by sexual partners and sexual relations tend to be based on exchange, bar workers cannot simply be equated with prostitutes. Some have a regular partner and the odd casual partner while others may have large numbers of casual contacts. Regular partners are almost always married and often itinerant. The distinction between regular and casual partners is important and based on the nature and extent of financial support. It is also related to condom use and therefore to risk. Women claim to be able to demand condom use from casual partners but not from regular partners. Although women claim that regular partners can be trusted, they nonetheless admit feeling at risk of AIDS and STDs from these same partners. PMID- 9337886 TI - STD/HIV/AIDS knowledge, beliefs and practices of traditional healers in Botswana. AB - The study investigated knowledge, beliefs, practices and experiences of traditional healers in relation to sexually transmitted diseases (STDs), HIV and AIDS. Traditional healers see about 70% of the African patients, with all kinds of ailments. The advent of HIV/AIDS and the introduction of home-based care in most African countries has increased the case-load of many traditional healers and increased the risk of contact with people living with HIV/AIDS. To protect themselves and their clients they need the right information on HIV/AIDS. Most traditional healers use their bare hands as a diagnostic tool and to apply topical medicine. Many traditional healers also utilize their mouths to suck blood from their patient's body as part of disease management. Most of the patients who are discharged from hospitals on home-based care usually end up at the traditional healer as relatives seek a second opinion or simply because they disagree with the diagnosis of incurable disease. This exposes traditional healers to HIV/AIDS. The study showed that traditional healers have some practices and beliefs, such as the use of the mouth for sucking blood (blood letting), use of sharp instruments which is risky behaviour and the belief that HIV/AIDS is not a new disease. Further most of the traditional healers did not have adequate and in some cases correct information on HIV/AIDS. A few even believed they could cure AIDS as it has always been a disease they have been dealing with and were adamant it is not a new disease. Rapport between traditional healers and scientific medical personnel is essential for an effective and successful HIV/AIDS prevention and control programme. PMID- 9337887 TI - Widely varying HIV prevalence and risk behaviours among the ethnic minority peoples of northern Thailand. AB - We compared HIV prevalence, risk behaviours, and social and sexual norms among nine ethnic minority (Hilltribe) groups in northern Thailand. Communities were selected on the basis of size, ethnicity, development level and geography. Subjects (15-45 years) were stratified by gender and selected by household using two-stage randomization. Forty volunteers were identified in each of 27 villages. Participation was voluntary and informed consent was obtained. HIV infection status was determined using ELISA and Western Blot on saliva samples. Risks for HIV were measured with structured interviews using local languages. Overall HIV prevalence was 23/1080 (2.13%) with an equal male-female ratio. HIV prevalence rates were: Shan 8.75%, Akha 5.0%, Yao 5.0%, Thin 1.25%, Hmong 0.63%, Lahu 0.63%, Lisu 0.63%, Karen 0, and Pa-Long 0. Sex worker use was an HIV risk for men (p = 0.0001), but injecting drug use was not; for women, having been a sex worker was a significant HIV risk (p < 0.0001). HIV rates, social norms and sexual behaviour varied considerably among ethnic groups, as did attitudes toward commercial sex work and use. HIV prevention needs to target the Shan, Akha, and Yao communities, and to focus on reduction of brothel work and use. PMID- 9337888 TI - HIV-seropositive men who engage in high-risk sexual behaviour: psychological characteristics and implications for prevention. AB - A minority of people who test HIV seropositive continue to engage in sexual behaviour that places their partners at high risk for HIV infection. However, little is known about factors that contribute to sexual risk behaviour among. HIV seropositive men. In this study, HIV-seropositive men participating in substance abuse support groups and HIV prevention programmes (n = 223) completed measures of demographic characteristics, sexual behaviour history, sensation-seeking (the propensity to seek optimal stimulation), and sexual compulsivity (persistent sexual preoccupations). Twenty-six per cent of the sample reported having recent multiple unprotected sexual intercourse partners. Across support group and prevention programme participants, men with multiple unprotected partners reported greater sexual compulsivity than men with one or no unprotected partners, but groups did not differ in terms of sensation-seeking. Results suggest that intensive therapeutic interventions are needed for a relatively small number of people who may contribute significantly to the HIV epidemic. PMID- 9337890 TI - Cross-sectional survey of hospital paediatric HIV/AIDS care in Catalonia, 1992. Hospital Contributors. AB - A cross-sectional survey to assess the impact of the paediatric HIV/AIDS epidemic on the hospital-based health care system was performed in state-financed hospitals in Catalonia during 1992, raising issues of relevance today. Out of the 27 hospitals contacted, 20 responded. A considerable proportion of the health care to children with an HIV-related condition was provided by four hospitals. The average length of stay of the 176 HIV patients who were admitted was 10.8 days; these patients were admitted to the hospital twice a year on average. Nearly half of the out-patients who attended with an HIV-related condition were either seropositive without a confirmed diagnosis of an HIV infection (class P-0) or seronegative. Thirteen per cent of the overall admissions to paediatric day care hospitals were attributable to an HIV-related condition. By ownership status of the hospitals, HIV/AIDS paediatric in-patients of public hospitals generated the majority of admissions per patient per year, and had the shortest lengths of stay. Unlike the HIV/AIDS epidemic in adults, the magnitude and characteristics of the epidemic in children may not require the shift of hospital-based health care to primary health care. PMID- 9337889 TI - A new out-patient care facility for HIV-infected destitute populations in Paris, France. AB - In France, the entire population theoretically has access to health coverage, but in fact a section of the poorest population does not. Institutions have therefore been set up to provide medical care for the destitute. The objectives of this study were to describe the social characteristics of the HIV-positive destitute population attending an out-patient clinic providing free health care for the destitute in a Paris University Hospital, to compare their clinical epidemiological characteristics with those of non-destitute HIV-positive patients, and to evaluate the quality of their care. We performed a historical prospective study wherein a cohort of 115 HIV-positive destitute patients (defined as having no health coverage at their first consultation) was compared with a control cohort of 183 HIV-positive non-destitute patients attending the same clinic. Ninety-five per cent of the destitute patients had no stable employment, 32% had no source of income, 75% had no permanent residence and 27% were i.v. drug abusers. Fifty-nine per cent were foreigners, most of whom had legal residence papers and had been in France for more than 3 years. When comparing the control and the destitute groups, the latter had a three times greater risk of developing tuberculosis (RH = 3.2, CI 95% = [1.1-9.4]). Medical compliance, access to antiretroviral treatment and hospitalization were identical in both groups. No difference was observed in terms of occurrence of a new AIDS related disease during follow-up when full-blown AIDS before entry, CD4 count at entry and transmission group were taken into account in multivariate analysis. From the moment that destitute patients attended this adapted medico-social facility, their access to care was the same as, if not better than, that of the other patients. The development of out-patient medico-social facilities for HIV positive destitute patients must be a public health priority even for those countries theoretically providing generalized health coverage. PMID- 9337891 TI - Sexual Risk Cognitions Questionnaire: a reliability and validity study. AB - The Sexual Risk Cognitions Questionnaire was designed to assess the type and frequency of cognitions associated with unsafe sex. It consists of 22 core items (SRCQ-22) with six subsections. Each subsection consists of 8-12 items designed for specific subgroups defined by gender, sexual orientation and HIV serostatus. Data on reliability and validity are presented based on a sample of 344 respondents in London. Reliability for all sections of the questionnaire was high but sample numbers were relatively small for some subsections. Validity for the SRCQ-22 was assessed in terms of its relationship with self-reported sexual behaviour among men who have sex with men who made up the largest proportion (70%) of the sample. The results indicate that the SRCQ-22 is a reliable measure for assessing cognitions related to HIV risk sexual behaviour in this population and supports the view that they are important determinants of safe and unsafe behaviour. PMID- 9337892 TI - Encounters with the Israeli Health Service: impressions from a support group for people with HIV. AB - The article which follows documents one of the first short-term support groups for HIV-infected people (PWH) in Israel. The group enabled its members to cope in a positive way with issues of life and death and with the daily hardships that accompany their condition, and became an essential supporting factor in their lives. Four main areas of difficulty emerged in the group pertaining to PWH encounters with the health service, of which all may have detrimental effects on compliance with treatment: (a) sensitivity to the patient; (b) ensuring the patient's privacy; (c) consistency in procedures and in providing information; (d) communication between doctor and patient. The importance of recognizing these difficulties and the need for improvement are underscored. PMID- 9337893 TI - [Calcium intake and bone growth]. PMID- 9337894 TI - [Oxycephaly, a severe craniosynostosis. Apropos of a series of 129 cases]. AB - AIMS: The authors analyse a series of patients with oxycephaly in order to detail the definition of this craniosynostosis and its functional prognosis. PATIENTS AND METHODS: The medical records of 129 oxycephalic patients were reviewed. Skull X-rays, ophthalmologic examination, mental level assessment, intracranial pressure monitoring and CT scan were analysed. The more recent patients were also analysed by MRI. Operated on or not, the patients were followed-up, particularly as far as the mental evolution is concerned. Mean follow-up was 3 years 7 months. RESULTS: One third of the patients came from North Africa, where oxycephaly seems predominant. Mean age at diagnosis was 6 years. Past history of rickets was found in 15% of the patients. On X-rays, the vast majority of the patients presented with multisutural synostosis involving both coronal and sagittal sutures, and diffuse digital prints. At the first mental assessment, one third of the patients had an IQ below 80. Papilledema was found in 17%. The monitoring of intracranial pressure showed an increased pressure in almost two thirds of the patients. Sixty four percent of the patients with increased intracranial pressure had a normal fundoscopy. Out of 16 patients explored by MRI, 12 had a Chiari I malformation. Postoperatively, all papilledemas disappeared, and the intracranial pressure returned to normal in all cases with preoperative increased intracranial pressure. The mental level seemed to stabilize, the mean postoperative IQ being strongly correlated with the preoperative level. In non-operated patients, the mental level worsened significantly. CONCLUSIONS: Oxycephaly is a late-appearing craniosynostosis, with a high risk of ophthalmologic and mental complications. Based on the present series, the operation seemed effective in preventing these complications. PMID- 9337895 TI - [Characteristics of patients and use of resource in French pediatric intensive care units. Le groupe francophone de Reanimation et urgences pediatriques]. AB - Evaluation of case-mix and resource consumption in pediatric intensive care units (ICU) is required. AIMS: This study describes the patterns of pediatric ICU resource consumption, determines the impact of primary clinical characteristics (particularly severity of illness) on resource utilization, and analyses medical efficiency with the frequency of inappropriate stays in French pediatric ICU. METHODS: Prospective study in nine French volunteer multidisciplinary pediatric ICUs from December 1993 to April 1994. Premature neonates were excluded. Resource consumption was expressed using the Omega system and length of stay, from which total Omega per admission, and average daily Omega (total Omega/length of stay) were obtained. RESULTS: Seven hundred and twelve patients were eligible. Twenty five percent were full-term neonates, 30% infants (1 month to 1 year), and 45% children. Surgical patients constituted 22% of the population. A chronic disease was present in 45% of infants and children. Immunodeficiency was present in 10% of patients. The median length of stay was 4 days (range: 1-155). The mean Omega scores per admission were: total Omega = 92 +/- 124, Omega/day = 14 +/- 9. Sixty four percent required mechanical ventilation and 37% during more than 2 days. Forty-two percent had a central venous access, and 23% an arterial line. The resource consumption was greater in non-survivors, surgical patients, neonates, and immunosuppressed patients. The mean PRISM score was 9 +/- 9. Mortality was 13%. The Omega/day and the PRISM score correlated. The frequency of inappropriate stays was 7.6% and accounted for 1.0% of the overall Omega activity. CONCLUSIONS: The results of this study can be used for interinstitutional comparison and a broader appraisal of pediatric intensive care. They illustrate the relationship between severity of illness and resource consumption. French pediatric ICU efficiency seems to be high, as compared to North American and Dutch results. PMID- 9337896 TI - [Traumatic pharyngoesophageal perforation in newborn infants]. AB - BACKGROUND: Neonatal perforation of the esophagus appears to occur rarely and often can mimic esophageal atresia. This report presents 12 cases of pharyngoesophageal perforation with a review of the literature. PATIENTS: From 1980 to 1995, we treated 12 infants for pharyngo-esophageal perforation. Ten infants were pre-term, seven of them weighing less than 1,500 g. Five infants had severe respiratory distress. Four infants had repeated attempt on intubation of the airway and eight infants had a routine postpartum suctioning and gastric aspiration. On plain X-ray, a large right pneumothorax was observed in three cases and the nasogastric tube deviated widely from its expected course in three cases. Four infants underwent contrast esophagography and three infants esophagoscopy. In five cases esophageal atresia was the initial diagnosis. Five infants underwent a thoracotomy. A gastrostomy was performed in one case. The six remaining neonates were treated non-operatively: broad spectrum antibiotics, total parenteral nutrition, and silastic nasogastric tube. Follow-up was uneventful in five cases. One infant with an esophageal stricture underwent instrumental dilatation. Bronchopulmonary dysplasia occurred in two cases and necrotizing enterocolitis in one. Two infants died. CONCLUSION: Iatrogenic perforation remains a difficult diagnosis. Clinical findings, plain chest x-ray and oesophagography are helpful. Surgery can be completely avoided in most instances. Infants with low birthweight and prematurity are most at risk. PMID- 9337897 TI - [Epidemiological estimation of the effectiveness of the pertussis vaccination during outbreaks in a community. Les pediatres et bacteriologistes du reseau RENACOQ]. AB - BACKGROUND: With the recent pertussis vaccine trials, the efficacy of acellular vaccines is now well known, estimated at 85% for multicomponent vaccines. On the other hand, the estimates of whole cell vaccines efficacy varies from 36% to 98% with the different vaccines used. We evaluated the field effectiveness of the French whole cell pertussis vaccine during outbreaks in schools and centers for disabled children. METHODS: Four limited outbreaks between 1993 and 1995 were investigated using a retrospective cohort study design. Vaccine effectiveness (VE) was assessed for the following case definition: clinical pertussis, laboratory-confirmed pertussis, epidemiologically confirmed pertussis (documented contact with a laboratory confirmed case). Immunization history was obtained by reviewing the child health record book. Effectiveness of a whole vaccination (four injections) and of a partial vaccination (one to three injections) were estimated as 1-(attack rate among vaccinated/attack rate among non-vaccinated). RESULTS: A whole immunization conferred good protection against pertussis with an estimated VE higher than 92% in three surveys, lower in the fourth survey (84%) in which antibiotic prophylaxis was set up very rapidly. A partial immunization conferred a mild protection (median: 60%). CONCLUSIONS: These results are consistent with a previous report about the effectiveness of this whole cell vaccine using the screening method in a hospital network survey in France. In the same way, a large efficacy trial in Senegal comparing it with an acellular bivalent vaccine estimated its efficacy at 96%. This high efficacy together with a satisfactory vaccine coverage leads to the current epidemiological profile of pertussis in childhood in France: majority of cases occurring before 6 months of age, limited outbreaks in school children, many of whom being unvaccinated or partially vaccinated. PMID- 9337898 TI - [Value and limits of selective bronchial obstruction in neonatal unilateral interstitial emphysema]. AB - BACKGROUND: Two methods of selective ventilation have been used for treating severe localized pulmonary emphysema in the neonates: controlateral selective intubation and selective bronchial obstruction. CASE REPORTS: Three neonates with acute respiratory distress required respiratory support that was complicated by development of severe localized pulmonary interstitial emphysema of the right lobe (two cases) and the middle lobe (one case). Selective bronchial obstruction with a Swann Ganz catheter SF was tentatively made: in one case, improvement was moderate and transitory, requiring middle lobectomy. The localized emphysema disappeared within 3 days in the two other cases but a localized emphysema appeared in the controlateral lung in one of them, requiring left inferior lobectomy because the ineffectiveness of selective intubation or selective obstruction. CONCLUSION: Selective bronchial obstruction may fail but this easy and well tolerated method should be tried in severe localized emphysema, specially in those patients who cannot be ventilated with high-frequency oscillation. PMID- 9337899 TI - [Benign reflex myoclonic epilepsy in infants]. AB - BACKGROUND: Myoclonic epilepsy of infancy are seldom benign. CASE REPORT: A 25 month old girl developed myoclonic jerks either spontaneously either as reflex responses to auditory and tactile stimuli, such as sudden touching of the face or trunk from the age of 4 months. The jerks disappeared after valproate therapy. Neurological examination was normal with a follow-up of 9 months. CONCLUSION: This condition resembles that described in 1995 by Ricci et al. In must be differentiated from other myoclonic epilepsies of infancy, reflex epilepsies and hyperekplexia. It could be the earliest from of idiopathic generalized epilepsy. PMID- 9337900 TI - [A familial case with generalized resistance to thyroid hormones]. AB - BACKGROUND: The syndrome of generalized resistance to thyroid hormones is more frequent than was thought. CASE REPORTS: A 13-year old girl was examined for her short stature. Evaluation of her thyroid function showed increased levels of IT3 and IT4 and normal value of TSH; she also had mosaic Turner's syndrome. Her cousin was rapidly diagnosed as suffering from the same syndrome because of moderately high thyrotropic levels found during neonatal screening; this syndrome was confirmed by molecular biology tests. Five generations of this family were identified as being affected with a pattern indicating autosomal dominant inheritance. CONCLUSION: The clinical manifestations of familial generalized resistance to thyroid hormones vary but this syndrome is easy to biologically confirm. The importance of diagnosing affected children as early as possible should be emphasized, as in such cases their development must be closely monitored particularly where their growth and neurodevelopment are concerned. PMID- 9337901 TI - [Ischemic cerebral vascular accident caused by vertebral artery dissection]. AB - BACKGROUND: Strokes due to vertebral artery lesions are not yet well known in children. CASE REPORT: We report on a case of post-traumatic vertebral artery dissection responsible for ischemic stroke in a 8-year old boy. Headache, vomiting and a brief loss of consciousness were the main initial signs. Neurological examination showed a locked-in syndrome. Cerebral imaging revealed lesions in cortical cerebellar hemisphere, cerebral pedoncular and protuberance. An arteriogram performed on day 10 showed left vertebral artery occlusion at C2 levels consistent with vertebral dissection. Antiagregants treatment was given. Neurological recovery was good. Pertinent clinical data of 24 children who had strokes due to a vertebral artery dissection are analysed. CONCLUSION: Vertebral artery dissection is presently a well-known cause of childhood strokes. Benefits from anticoagulants are now established. PMID- 9337902 TI - [Allergy to snails and mites in children]. AB - BACKGROUND: Anaphylactic reactions after consumption of snails by patients sensitized to house-dust mites have been reported several times. CASE REPORT: Two 8- and 10-year old children sensitized to house-dust mites developed Quincke's oedema after eating snails. Immunoallergologic investigations including pricks test, labial test, IgE Rast confirmed associated allergy between snails and house dust mites. CONCLUSION: Considering the potential severity of anaphylactic reactions, it is necessary to warn children allergic to house dust mites and their parents about the high risk of associated allergy with snails. PMID- 9337903 TI - [Munchausen syndrome by proxy]. AB - Munchausen syndrome by proxy is a form of child abuse presenting as a disease produced or simulated by a parent, the mother in most cases. Its diagnosis is uneasy because of its miscellaneous and unusual clinical presentation and of the misleading apparently normal attitude of the parents. Physicians may participate in the abuse by insistently looking for diagnostic and therapeutic measures, therefore contributing to the significant mortality of the syndrome. It is therefore important that physicians consider Munchausen syndrome in any ambiguous situation in order to protect the child by an early diagnosis. PMID- 9337904 TI - [Treatment of sleep disorders in children]. AB - Sleep disorders require careful evaluation, precise diagnosis and a systematic search for a cause before considering treatment. Hypnotics must never be prescribed directly and when necessary it must be used precautionally during a short period of time, as little is known on their long term effects in children. In addition some psychotropic drugs may lead to tolerance and addiction. Educational and behavioural therapy together with sleep hygiene, have proven to be efficient in most situations. Awaking stimulants are indicated in primary hypersomnia. PMID- 9337906 TI - [Treatment of malignant liver tumors in children: evaluation and prospects]. AB - Rather discouraging in the past, treatment of malignant tumors in children allows today a 75% cure rate for hepatoblastoma. Complete surgical resection remains the ongoing basis of the treatment, but the main advances are due to more efficient chemotherapy protocols using cisplatin, to an improvement in imaging procedures, to modern techniques of anesthaesia, to aggressive surgery and treatment of metastases, and finally to liver transplantation when the extension of the tumor precludes total resection in the absence of metastasis. The management of children with malignant tumors should be performed in selected centres participating in collaborative protocols, therefore providing the best oncological and surgical standards and the possibility of liver transplantation if necessary. PMID- 9337905 TI - [Suicidal behavior in adolescents in Switzerland: role of physicians]. AB - INTRODUCTION: In most industrialized countries, suicide represents the second leading cause of death among adolescents. Swiss teenagers exhibit one of the highest death rates by suicide in Europe; however, the prevalence of suicidal conducts (suicidal ideas, projects and attempts) in Switzerland is not known. OBJECTIVES: To assess the prevalence of suicidal conducts among Swiss adolescents and to compare these figures with available data from other countries. To establish how suicidal adolescents use health services in comparison with non suicidal adolescents. METHODS: Bivariate analyses have been performed using data from the "SMASH" study (Swiss Multicenter Adolescent Survey on Health), a national survey on the health and lifestyles of 9,268 15 to 20 years in-school youth (3,993 girls et 5,275 boys). Within a self-administered anonymous questionnaire of 80 items, five specific questions focused on suicidal conducts and lead to the distribution of respondents in four groups: no suicidal concern, suicidal ideas, suicidal plans, suicide attempt. RESULTS: Regarding the last 12 months, 5,144 teenagers (55.5%) report no suicide preoccupation at all; 2,376 (25.6%) report suicide ideas only, 1,366 (14.7%) report suicidal projects and 274 (3%) report suicide attempts. Only 40% of respondents who report a suicide attempt have talked about it to someone in their circle of family or friends. Moreover, less than 20% have spoken of their suicide with a psychologist, and although they see physicians as often as the rest of the sample, only 10% have discussed their attempt with one of them. In comparison with those free of suicidal concerns, suicidal adolescents report significantly more health problems and concerns: they feel more often tired or depressed and use medication more often. They also seem to exhibit more deviant behaviors like alcohol and drug use or runaway. CONCLUSION: Suicidal conducts, especially suicide attempts are much more prevalent among Swiss adolescents, but a small minority is acknowledged and treated by the medical profession. Physicians should be aware of this pathology and better trained both in term of detection and treatment. PMID- 9337907 TI - [Radiological case of the month. Unilateral genu valgum revealing Ollier's disease]. PMID- 9337908 TI - [Current methods for the localization of parathyroid adenoma in children]. PMID- 9337909 TI - [Indications for and technique of intensive phototherapy of full-term newborn infants]. PMID- 9337910 TI - How robust is the concept of post-traumatic stress disorder? PMID- 9337911 TI - The role of the consultant in communicable disease control. PMID- 9337912 TI - Interventional neuroradiology techniques in neurovascular disease. PMID- 9337913 TI - Resuscitation. 1: Basic life support. AB - Basic life support (BLS) is a method of sustaining vital functions in a person who has collapsed and is unconscious, frequently with a cardiac arrest but sometimes with respiratory arrest, choking or other cause. BLS is a vital link in the 'chain of survival' for these critically ill people. PMID- 9337914 TI - Anaesthetic management of severely injured patients: general issues. PMID- 9337915 TI - Clinical epidemiology of coronary heart disease in the UK. PMID- 9337916 TI - Myocardial infarction: early diagnosis. PMID- 9337917 TI - Risk stratification of patients post-myocardial infarction. PMID- 9337918 TI - Primary angioplasty for the treatment of acute myocardial infarction. PMID- 9337919 TI - Pathophysiology and management of portal hypertension. 1: Variceal haemorrhage. AB - Portal hypertension occurs secondary to a combination of increased resistance to portal venous flow and increased splanchnic inflow to the portal venous system. The main clinical complication is gastrooesophageal haemorrhage from which mortality remains high at approximately 40%. PMID- 9337920 TI - Psychiatric problems associated with alcohol misuse and dependence. AB - Psychiatric comorbidity is common in individuals with alcohol problems and has a significant effect on the outcome of alcohol problems. Problem drinkers should therefore be screened for psychiatric disorders and have access to appropriate treatment. Psychiatric comorbidity should be taken into account in the planning and development of treatment services for alcohol problems. PMID- 9337921 TI - Prospects for respiratory gene therapy. AB - Over the last 5 years a number of articles have been published on gene therapy for a range of diseases. The initial tone of a number of these articles evoked high expectations, particularly from the public. Over the last 2-3 years data from these studies have begun to filter through and, perhaps not surprisingly, initial problems have been encountered, resulting in a recent spate of bad press for gene therapy. This article aims to examine where the prospects for respiratory gene therapy lie in the face of these mixed reactions. PMID- 9337922 TI - Survival from cancer: local control and radiotherapy. AB - Adjuvant radiotherapy is considered to be highly effective in maintaining local control but is widely perceived to confer no survival advantage in the management of solid tumours. However, recent adjuvant radiotherapy studies are beginning to show survival improvements in parallel with improvements in loco-regional disease control. PMID- 9337923 TI - The notes in the cupboard: the question of intellectual honesty in neurosurgery. PMID- 9337924 TI - Lumbar disc herniations in children: a long-term clinical and magnetic resonance imaging follow-up study. AB - To determine the long-term outcome of 12 youthful patients with lumbar disc herniation, who, at the time of surgery, were 15 years old and younger (mean age at operation 14.3 years), we assessed their current clinical condition (mean follow-up time 6 years) with a questionnaire inquiring about symptoms and disability, and radiologically with an MRI of the lumbar spine. Clinically, only five patients (40%) were totally asymptomatic and seven patients (60%) had recurring symptoms, both and disability. On MRI, seven patients (60%) had persistent stenosing changes at the operated disc levels and eight patients (65%) also had disc degeneration at other lumbar levels. Despite the symptoms and quite severe radiological findings, the long-term outcome was assessed as good or moderate in eleven patients (90%). As far as comparisons are reasonable, our results appear somewhat less favourable than those in two previous paediatric series, but they agree with those in two recent large series of adults. PMID- 9337925 TI - Central neurocytoma: a clinico-pathological study of five cases. AB - Central neurocytoma (CN) is a rare, benign tumour of neuronal differentiation which affects young patients and is generally found in the lateral or third ventricles. Its radiological features are non-specific and, in the past, these tumours were confused with other intraventricular lesions. Only recently, thanks to their characteristic features on immunohistochemistry and electron microscopy, have they been recognized as a separate entity. We present the clinico pathological features of five cases of CN treated at our Institution between 1986 and 1994. The importance of diagnostic suspicion, total microsurgical excision and the role of radiotherapy is discussed. PMID- 9337926 TI - Acoustic neuroma and the eye. AB - A retrospective survey is presented of the case records of 138 patients who had undergone operative treatment for acoustic neuroma. The nature and incidence of ophthalmic features prior to and following surgery is documented. The study covers 12 years in two regional neurosurgical centres, under the care of six different neurosurgeons, one otolaryngologist and nine ophthalmologists. Of the 138 records examined, 61 patients (44%) required lid surgery of one variety or another. 18 (13%) developed minor superficial exposure keratopathy, 13 (9%) developed corneal opacification or clouding, two had recurrent infective abscesses and four developed optic atrophy. The development of corneal complications strongly correlates with the presence of documented preoperative fifth nerve involvement. Postoperative oculomotor cranial nerve palsies were seen in 10 patients (7%). Ophthalmologists should be involved in the perioperative management of these patients and certainly before irreversible corneal damage has occurred. PMID- 9337927 TI - Outcome and prognostic factors in the surgical treatment of spinal dural arteriovenous fistulas. A long-term study. AB - From a total of 78 patients surgically treated for a spinal dural arteriovenous fistula, in whom long-term follow-up was available, 25 have been reviewed. Their outcome at different postoperative stages and the long-term prognostic factors is discussed. Excision or coagulation of the nidus, as well as disconnection of the draining vein offers better results in the long-term than clipping of the draining vein alone. Prolonged duration of symptoms and poor functional status before surgical treatment adversely affect the long-term outcome. Age at the time of surgery did not influence outcome in this study. In the long-term all patients tended to show a moderate, but definite functional decline. This is unlikely to be due entirely to recanalization and may represent a more generalized haemodynamic abnormality of the cord together with the effects of age on the original damage. PMID- 9337928 TI - Idiopathic syringomyelia and the importance of occult arachnoid webs, pouches and cysts. AB - Syringomyelia is the condition in which longitudinal cavities are found within the spinal cord. The use of drainage procedures has been widely practised with good short term results. However, the long-term results in some large series have been less favourable. There are many associated conditions and in most forms a blockage to the normal flow of CSF, either at the foramen magnum or in the spinal canal, can be identified. Most surgeons would now direct their efforts to the establishment of normal CSF flow rather than a shunting procedure. In a certain group of patients, even with the advent of sophisticated MRI, no associated abnormality or CSF block is easily identified. This type of syringomyelia is often termed idiopathic. We report 10 patients with symptomatic syringomyelia without readily recognized predisposing factors. In eight patients preoperative myelography revealed a block to the flow of contrast compatible with subarachnoid obstruction. Eight patients underwent laminectomy and division of the obstructing arachnoid webs. Five experienced good improvement and three only moderate improvement. Two of the patients underwent syrinx shunting procedures only, which resulted in a worsening of their symptoms. At operation one patient was found to have an arachnoid cyst. We believe that patients with idiopathic symptomatic syringomyelia may need myelography to identify such arachnoid abnormalities. Subsequent surgery should be directed at the establishment of normal CSF flow by laminectomy and excision of the offending lesion. PMID- 9337929 TI - The transconjunctival microsurgical approach to the orbit: recent experience in 22 cases. AB - We report a 5-year follow-up of 22 consecutive patients, who underwent transconjunctival management of their orbital lesions without muscle dissection. The presentation, natural history, management, surgical appraisal of the transconjunctival approach and its indications are discussed. At follow-up, 21 of the patients showed excellent cosmetic and functional results. Our experience suggests that this approach remains a useful modality, with good intraoperative visibility and minimal postoperative scar formation, for managing selected patients with space-occupying lesions located in the inferior medial and basal compartment of the orbit. Not suitable for the transconjunctival approach are deep intraconal lesions (in the orbital apex) and extraconal superior lesions. Rare complications of transconjunctival approach have included temporary eye muscle injury with ophthalmoplegia postoperatively. More recently, the use of the transconjunctival approach has allowed surgeons to reduce cosmetic failures, functional deficits and deformities of the orbit. Because of its low risks, the absence of postoperative bleeding, and the limited hospitalization and immobilization of the patients, the transconjunctival approach is a successful procedure especially in elderly patients with intercurrent disease. PMID- 9337930 TI - Good practice in the management of adults with malignant cerebral glioma: clinical guidelines. Working Group, Royal College of Physicians. AB - This paper proposes guidelines for good practice in the management of adults with malignant cerebral glioma. These guidelines were developed by a working group comprising representatives of the medical specialties involved in patient care, specialist nursing staff, purchasers, charitable bodies, and patient and relative representatives. Both the research literature on the effectiveness of medical intervention, and the views of patients and relatives about the care they had received were considered. The document proposes a consensus view about ways to improve patient care and considers several stages of the illness and its care: I, the diagnostic phase; II, deciding on an appropriate treatment plan; III, the organization of follow-up services; IV, the management of transitions from hospital to community settings; and V, purchasing care for patients with malignant brain tumours. An audit package derived from the guidelines is available which will enable staff within a treatment centre to compare their practice against these standards. A final section suggests topics which require further research, and sets out the core requirements for studies that will help answer questions about treatment and the benefits for patients in terms of improved quality of life. PMID- 9337931 TI - Stereotactic multiple arc radiotherapy. V: Primary treatment of discrete low grade glioma. AB - Two, discrete and spherical low grade gliomas, both demonstrating homogenous signal characteristics on MRI, were treated by radical radiosurgery. Both demonstrated the central low intensity signal alterations that are now well described as occurring after radiosurgery of cranial neuroma. Growth arrest occurred in both cases by 2 years, followed by slow shrinkage in case 2 with the longer follow-up. Both patients are well at 2 and 5 years after treatment. These observations are discussed with regard to the role of radiosurgery in glioma therapy. PMID- 9337932 TI - A simple technique to limit ICP catheter infection. AB - Infection of intracranial catheters is a common complication of ICP monitoring. The introduction of a simple technique of ensheathing the entire length of the external segment of the catheter in a sterile plastic sheath resulted in a decreased infection rate. In the study year, one of 78 patients developed catheter-induced meningitis, compared with seven of 64 patients in the year prior to the introduction of the protective plastic sheath. The use of a plastic sheath resulted in a low rate of infection. PMID- 9337933 TI - Ossified glomus jugulare tumour: case report. AB - A huge, ossified and highly vascular glomus jugulare tumour in a 19-year-old boy was radically and successfully resected. External carotid artery embolization and intermittent internal carotid artery trapping during surgery were the principle methods employed to control the operative blood loss. Extensive petrous bone resection, and adequate and wide exposure were necessary. The case and the operative steps in this unusual and difficult surgical problem are discussed. PMID- 9337934 TI - Mutism, oropharyngeal apraxia and dysarthria after posterior fossa tumour excision. AB - Mutism and oropharyngeal apraxia are unusual complications of surgery on the cerebellum. They usually occur in children undergoing surgery for midline cerebellar tumours. Adults are rarely affected. The pathophysiology of the syndrome, which is reversible, is uncertain with possible involvement of vermian and paravermian structures. Two patients--one child and one adult--who developed mutism after cerebellar surgery are presented. PMID- 9337935 TI - Value of early postoperative plain radiography following BOP fusion in cervical spinal surgery. AB - After anterior decompression and interbody fusion with biocompatible osteoconductive polymer (BOP) for cervical degenerative disease it is common practice to perform early radiography to evaluate the results, as a questionnaire sent to 136 consultants neurosurgeons in UK demonstrated. Radiographs of 39 patients following the procedure were analysed and it was concluded that the practice is of no value. PMID- 9337936 TI - The association of cavernous and venous angioma. Case report and review of the literature. AB - We report the case of a patient with progressive seizures caused by a cavernous angioma in association with a venous angioma in the right parietal lobe. The radiological findings, the pathogenesis of this association and the importance of surgical treatment of a cavernous angioma with conservative treatment of a venous angioma are discussed. PMID- 9337937 TI - Prolactin-secreting carcinoma of the pituitary: clinicopathological and immunohistochemical study of a case with intracranial and intraspinal dissemination. AB - A 32-year-old female patient with a primary adenohypophyseal neoplasm that rapidly progressed to a fatal outcome is presented. The time interval between her admission to the hospital and her death was 3 months. Despite dopamine agonist therapy, local invasion as well as frontal and spinal cord metastases at Th 10-12 region developed, and four surgical resections were performed. The serum prolactin levels were high. Both the primary pituitary tumour and all the metastatic tumours had the same histological findings and immunohistochemical reactions. Each was composed of pleomorphic chromophobic cells with enlarged nuclei. Mitoses and necroses were frequent. Immunostains revealed prolactin in the tumour cells. A literature review revealed that in most of the pituitary carcinomas as in our case hyperprolactinaemia did not respond to medical therapy and the histopathological appearance of the tumour has not correlated with the aggressive behaviour of the tumour. It may therefore be considered that at least some of the cases with metastases in prolactin secreting pituitary carcinomas could be the result of hyperprolactinaemia itself. PMID- 9337938 TI - Avascular necrosis secondary to postoperative steroid therapy. AB - Hypothalamic and pituitary tumours may present with vague symptoms owing to excess or lack of hormone production, including diabetes insipidus. Corticosteroids are commonly employed to limit cerebral oedema and at much lower doses to treat secondary hypocorticalism. Continuation of steroids at inappropriately high doses predisposes to the development of avascular necrosis as in the case we describe in a young woman of 34 years. This is a potentially preventable crippling disorder. When prescribing steroids the lowest effective dose should be used. PMID- 9337939 TI - Intradiploic meningocoele in a 16-year-old girl. AB - Osteolytic lesions of the skull have many causes, but magnetic resonance imaging may immediately distinguish cerebrospinal fluid intradiploic lakes from other causes. A case of a 16-year-old girl with such a lesion in relation to the sagittal sinus is presented and its aetiology is discussed. PMID- 9337940 TI - Establishing cost-effectiveness of atypical neuroleptics. PMID- 9337941 TI - Maternal stress or anxiety in pregnancy and emotional development of the child. PMID- 9337942 TI - Long-term cannabis use and mental health. PMID- 9337943 TI - Pharmacokinetic interactions involving clozapine. AB - BACKGROUND: Metabolism of clozapine is complex and not fully understood. Pharmacokinetic interactions with other drugs have been described but, in some cases, their mechanism is unknown. METHOD: Published trials and case reports relevant to the human metabolism of clozapine and to suspected pharmacokinetic interactions were reviewed. RESULTS: Metabolism of clozapine appears to be largely controlled by the function of the hepatic cytochrome p450IA2 (CYPIA2). Compounds which induce CYPIA2 activity (carbamazepine, tobacco smoke) may reduce plasma clozapine levels. Inhibitors of CYPIA2 (caffeine, erythromycin) have the opposite effect. Drugs which inhibit the hepatic cytochrome p4502D6 (CYP2D6) have also been reported to elevate plasma clozapine levels. The mechanism of this interaction is unclear. CONCLUSIONS: The co-administration of clozapine and compounds reported to alter its metabolism should be avoided where possible. A host of other interactions can be predicted and so caution should be exercised when co-administering drugs which affect the function of CYPIA2 and CYP2D6. The pharmacokinetics of clozapine require further investigation so that its safe use can be assured. PMID- 9337944 TI - Lithium: evidence reconsidered. PMID- 9337945 TI - Lithium: balancing risks and benefits. PMID- 9337946 TI - Cost-effectiveness of clozapine. A UK clinic-based study. AB - BACKGROUND: Schizophrenia is highly expensive in calculable and incalculable costs. Measures which impact the cost in the most severely affected are likely to produce the greatest cost reductions. Studies regarding clozapine in the USA have demonstrated clear cost-effectiveness, despite the high prescription costs. There are no prior UK studies. METHOD: We performed a cost-effectiveness analysis comparing the three years prior to commencing clozapine to the period following establishment of clozapine treatment (mean 36.4 months) for 26 patients with chronic schizophrenia or schizoaffective disorder. RESULTS: There was a significant improvement in all clinical ratings applied (and a mean net saving of ponds 3768 per annum). The cost-effectiveness of clozapine was double that of conventional neuroleptics (15.2 pre-, 33.0 post-clozapine, P < 0.005). CONCLUSIONS: As a naturalistic study our data provide valuable information on the cost-effectiveness of clozapine in the UK. Our methodology could be applied in a community setting or in the study of another atypical neuroleptic. PMID- 9337947 TI - Acute and one-year outcome of a randomised controlled trial of brief cognitive therapy for major depressive disorder in primary care. AB - BACKGROUND: The consensus statement on the treatment of depression (Paykel & Priest, 1992) advocates the use of cognitive therapy techniques as an adjunct to medication. METHOD: This paper describes a randomised controlled trial of brief cognitive therapy (BCT) plus 'treatment as usual' versus treatment as usual in the management of 48 patients with major depressive disorder presenting in primary care. RESULTS: At the end of the acute phase, significantly more subjects (P < 0.05) met recovery criteria in the intervention group (n = 15) compared with the control group (n = 8). When initial neuroticism scores were controlled for, reductions in Beck Depression Inventory and Hamilton Rating Scale for Depression scores favoured the BCT group throughout the 12 months of follow-up. CONCLUSIONS: BCT may be beneficial, but given the time constraints, therapists need to be more rather than less skilled in cognitive therapy. This, plus methodological limitations, leads us to advise caution before applying this approach more widely in primary care. PMID- 9337948 TI - Controlled trial of exposure and response prevention in obsessive-compulsive disorder. AB - BACKGROUND: Exposure and response prevention is considered a treatment of choice for obsessive-compulsive disorder (OCD). Yet there have been very few randomised controlled trials employing credible placebo conditions. This study compares exposure and response prevention with a general anxiety management intervention. METHOD: Eighteen patients meeting DSM-IV criteria for OCD were randomly assigned to either exposure and response prevention or anxiety management. Both treatments involved approximately 15 hours of therapy over a three-week period. RESULTS: There was a significant reduction in obsessive-compulsive symptoms following treatment with exposure and response prevention, while no change occurred in the control group. This was found to be statistically significant using a composite measure of OCD symptom severity, patient ratings of interference and therapist ratings of symptom severity. CONCLUSIONS: These findings suggest that the symptom reductions associated with behaviour therapy for OCD are a result of the specific techniques of exposure and response prevention, rather than non-specific aspects of the therapy process. General anxiety management techniques are not effective in the treatment of OCD. PMID- 9337949 TI - Incidence of schizophrenia in Nottingham. A comparison of two cohorts, 1978-80 and 1992-94. AB - BACKGROUND: Several studies have reported a decline of up to 50% in the incidence of schizophrenia over recent decades. We aimed to measure changes in the incidence and diagnostic patterns of first-episode psychosis by comparing two Nottingham cohorts, identified in two equal periods separated by 14 years. METHOD: Two prospectively ascertained cohorts of first-episode psychotic disorder were identified over the time periods 1978-80 and 1992-94. The earlier cohort was of the World Health Organization Determinants of Outcome of Severe Mental Disorder (DOSMD) ten-country study. The later cohort was obtained using similar methodology. Both groups were diagnosed using ICD-10 diagnostic criteria and age standardised incidence rates were compared. RESULTS: The standardised incidence rate for all psychotic disorders rose slightly from 2.49 to 2.87 per 10000 population per year, but the F20 classification fell significantly by over a third (1.41 to 0.87 per 10000 per year). The second study group (1992-1994) included a greater diversity of psychotic diagnoses compared with the first, in particular an increased proportion of acute and drug-related psychoses. CONCLUSIONS: Methodological considerations call for caution in interpreting such data, but we conclude that the significant fall in the narrowly defined diagnostic category of schizophrenia reflects a real change in the syndromal presentation of psychotic disorders. PMID- 9337950 TI - Increased rate of psychosis among African-Caribbeans in Britain is not due to an excess of pregnancy and birth complications. AB - BACKGROUND: It has been suggested that the increased rate of psychotic illness among African-Caribbeans living in Britain is due to an excess of pregnancy and birth complications (PBCs). METHOD: We therefore compared the frequency of PBCs in a group of White psychotic patients (n = 103) and a comparable group of patients of African-Caribbean origin (n = 61); the latter consisted of 30 first generation (born in the Caribbean) and 31 second-generation (born in Britain) individuals. RESULTS: White psychotic patients were more than twice as likely to have a history of PBCs as their African-Caribbean counterparts (odds ratio = 2.34, 95% confidence interval (CI) 0.88-6.47, P = 0.062). The same trend was observed among patients with a DSM-III diagnosis of schizophrenia (odds ratio = 1.65, 95% CI 0.56-4.97, P = 0.32). The rate of PBCs was similar among the first- and second-generation Caribbean psychotic patients. CONCLUSIONS: The increased rate of psychotic illness that has been reported among the African-Caribbean population in Britain is not due to an increased prevalence of PBCs. PMID- 9337951 TI - Incidence and risk factors for severe tardive dyskinesia in older patients. AB - BACKGROUND: Severe tardive dyskinesia (TD) represents a serious and potentially disabling movement disorder, yet relatively little is known about the incidence of and risk factors for severe TD. METHOD: We report the results of a longitudinal prospective incidence study of severe TD in 378 middle-aged and elderly neuropsychiatric patients. Psychiatric, neuropsychological, pharmacological and motor variables were obtained at intake and at regular intervals for 36 months. RESULTS: The cumulative incidence of severe TD was 2.5% after one year, 12.1% after two years, and 22.9% after three years. Individual univariable Cox regression analyses were conducted to identify demographic, psychiatric, motor and pharmacological predictors of severe TD. Results indicated that higher daily doses of neuroleptics at study entry, greater cumulative amounts of prescribed neuroleptic, and greater severity of worsening negative symptoms were predictive of severe TD. CONCLUSIONS: These findings suggest that conventional neuroleptics may be prescribed to older patients only when necessary and at the lowest effective dosage. Additional caution is recommended in patients exhibiting negative symptoms. PMID- 9337952 TI - Executive (frontal) dysfunction and negative symptoms in schizophrenia: apparent gender differences in 'static' v. 'progressive' profiles. AB - BACKGROUND: While executive (frontal lobe) dysfunction appears to be a core feature of schizophrenia, its relationship to psychopathology, age and duration of illness has yet to be explored systematically between the genders. METHOD: Executive dysfunction, positive and negative symptoms were evaluated in 27 male and 21 female in-patients who were unusually well-matched on numerous demographic and clinical measures. RESULTS: Measures of executive dyscontrol and negative symptoms were highly associated in both genders. However, while both executive dyscontrol and negative symptoms increased prominently with age/ duration of illness among women, no such relationship was evident among men. CONCLUSIONS: The similarly prominent levels of current executive dyscontrol and negative symptoms in male and female patients appear to have emerged via processes that differ fundamentally between the genders; among males these deficits appear to emerge and become 'locked in' earlier in the course of illness and to show little subsequent increase, while among females these same deficits appear to be less evident early in the course but to increase in prominence thereafter. PMID- 9337953 TI - Service provision for people with schizophrenia. I. Clinical and economic perspective. AB - BACKGROUND: The aim of this study was to provide information on patients current service use which could inform future decisions on service planning and resource allocation. METHOD: Individuals with a diagnosis of schizophrenia, who had received in-patient care in the previous five years, were identified from the Lothian Case Register. Information was obtained from 193 subjects. Patients' service use over a six-month period was examined. The costs incurred in service provision were determined. RESULTS: Patients differed markedly in their use of services. This was not found to be related to their mental state. Average care costs were high. In-patient care accounted for most of the overall expenditure. CONCLUSIONS: There is considerable variation in the services used by patients with schizophrenia and in the costs incurred in service provision. When planning services it is therefore important that detailed information on the patient population is available if resources are to be allocated cost-effectively. PMID- 9337954 TI - Service provision for people with schizophrenia. II. Role of the general practitioner. AB - BACKGROUND: This second report of a study of service provision for patients with schizophrenia describes patients' contact with general practice and general practitioners' (GPs') views of the mental health services. METHOD: A postal questionnaire was sent to the GPs, and patients' primary care records were examined. RESULTS: Data were collected on 131 subjects. The majority of patients (96) (73%) were in regular contact with their GP and were consulting for many different reasons; 27 (21%) were posing particular difficulties for the primary care team. GPs reported that 27 (21%) patients required additional support and that the care arrangements for 50 (38%) patients could be improved if alterations were made to the roles of the professionals already involved. CONCLUSIONS: GPs are central to service provision for patients with schizophrenia. Both additional resources and changes in working practices are required to improve patient care. The service implications of these findings are discussed. PMID- 9337955 TI - Integration between primary and secondary services in the care of the severely mentally ill: patients' and general practitioners' views. AB - BACKGROUND: Communication between secondary and primary care is an important aspect of continuity of care. We investigated communication between general practitioners (GPs) and psychiatric teams about a representative group of patients with severe mental illness (SMI). We also sought views on GP involvement in care from the patients and their GPs. METHODS: One hundred patients with SMI were randomly selected from those under the care of two psychiatric sector teams in inner London. The patients and their GPs were interviewed. RESULTS: GPs' knowledge about the care their patients received was limited. Most GPs perceived their role as providing physical care and prescribing. Few patients consulted GPs for mental health care. GPs perceived themselves as less involved in the care of Black Caribbean or Black African patients. CONCLUSIONS: Considerable discontinuities of care between secondary and primary care were identified. GP involvement in the care of patients with SMI appears limited. Better communication is necessary if care is to be shared. PMID- 9337956 TI - "Anybody's child": severe disorders of mother-to-infant bonding. AB - BACKGROUND: This paper describes severe, disorders of maternal affection and behaviour and suggests that there is an early process of mother-to-infant bonding which can go seriously wrong. METHOD: Forty-four self-selected women who had suffered from at least one episode of postnatal mental illness described an unexpected and often catastrophic failure to love one or more of their babies. RESULTS: These women reported absent affection, sometimes hate, rejection, neglect or impulses to harm, in relation to at least one of their children. These feelings often began immediately or very shortly after the birth, and with one exception, were specific to one child; such characteristics are best encapsulated by the term 'maternal bonding disorder'. Twenty-nine of the women were multiparae; first-borns were not significantly more likely to be the focus for such feelings. There was no direct evidence of predisposing maternal personality traits or previous experiences. Postnatal mental illness and recalled severe pain during labour were significantly associated with such disorders which, in their severe forms, did not occur in the absence of postnatal mental illness. CONCLUSIONS: The nature of the link between postnatal mental illness and disorders of maternal bonding remains unclear. Because, in multiparae, the disorder often 'missed' the first child, factors such as maternal personality traits or early childhood experiences cannot be regarded as sufficient causes. PMID- 9337958 TI - Long-term mortality after first psychiatric admission. PMID- 9337957 TI - Plasma noradrenaline response to electroconvulsive therapy in depressive illness. AB - BACKGROUND: Abnormalities of catecholaminergic function have been hypothesised to cause depressive illness. Plasma noradrenaline can be used as a marker of central noradrenergic activity. It is of interest to examine the change in resting plasma noradrenaline in patients with depressive illness over a course of electroconvulsive therapy (ECT) and relate this to their clinical state. METHOD: Patients referred for ECT who suffered from DSM-III-R major depressive disorder or dysthymia were recruited. Blood samples were taken before and after each treatment, during a course of ECT, to measure plasma noradrenaline and cortisol. Clinical ratings were carried out weekly during the course of ECT. RESULTS: Plasma noradrenaline fell significantly in those patients with melancholic/psychotic depressions but increased in those with non-melancholic depressive illness. There was a strong trend indicating that a fall in plasma noradrenaline was associated with improvement in depression ratings in the melancholic/psychotic patients only. CONCLUSIONS: Electroconvlusive therapy decreases plasma noradrenaline in melancholic/psychotic depressive illness and this shows a trend associated with clinical improvement. PMID- 9337959 TI - Long-term mortality after first psychiatric admission. PMID- 9337960 TI - Transcending barriers between religion and psychiatry. PMID- 9337961 TI - Gender and age at onset of schizophrenia. PMID- 9337962 TI - War pensions. PMID- 9337963 TI - Suicide and the cost-effectiveness of antidepressants. PMID- 9337964 TI - Crime, violence and schizophrenia. PMID- 9337965 TI - Anorexia and the overvalued idea. PMID- 9337966 TI - Genetics and psychiatry. PMID- 9337967 TI - Psychiatry, medicine and consultation-liaison. PMID- 9337968 TI - Dual diagnosis of severe mental illness and substance misuse: a case for specialist services? PMID- 9337969 TI - Opportunities for psychiatry from genetic findings. AB - BACKGROUND: The opportunities for psychiatry deriving from available or likely genetic advances are reviewed. METHOD: Clinical implications are considered in the context of both the misconceptions and benefits associated with relevant genetic findings. RESULTS: Misconceptions include that: heritability estimates have a 'true' fixed value; a high heritability means that environmental interventions will be ineffective; a high heritability within groups means that differences between groups will also be due to genes; genetic effects are determinative; 'genetic' means single abnormal genes; genes associated with disease must be bad and justify eugenic measures; gene therapy will be widely applicable; and genetic screening of the general population will be useful. The benefits include demonstrations that: both genes and environment have an ubiquitous influence; some prevailing diagnostic assumptions are mistaken; genes influence development; the effects of nature and nurture are not separate; and environmental effects tend to be person-specific. The potential value of molecular genetics lies in elucidation of causal processes as they apply to both brain systems and nature-nurture interplay; improving diagnosis and genetic counselling; and the development of improved pharmacological interventions. CONCLUSION: Advances in genetics will make a major impact on clinical psychiatry, and should bring practical benefits for both prevention and treatment. PMID- 9337970 TI - Closing the gap between research and practice. PMID- 9337971 TI - Psychiatry: evidence-based but still valve-laden. PMID- 9337973 TI - Closing the gap between research and practice. New evidence is required. PMID- 9337972 TI - Rational decision-making in psychiatry: evidence-based psychiatry is just the start. PMID- 9337974 TI - Psychiatrists and their patients: views on forms of dress and address. AB - BACKGROUND: Dress styles and forms of address vary among psychiatrists. METHOD: A semi-structured interview was administered to a sample of psychiatric in patients, and a questionnaire was sent to junior and consultant psychiatrists, to identify preferences for dress styles and terms of address. RESULTS: Forty-nine (71%) of the in-patient sample participated. A preference was found for smart attire and white coats. Of the 69 (80%) doctors returning questionnaires, the majority supported smart dress as the most appropriate attire. Most patients preferred to be called by their first name while addressing doctors by title and surname. Junior doctors preferred to use first names when talking to patients while almost all consultants used title and surname. Doctors of all grades liked to be called by their title and surname. CONCLUSIONS: Paying more attention to the way we present ourselves and interact at work may help to facilitate the therapeutic alliance. PMID- 9337975 TI - Helpfulness of interventions for mental disorders: beliefs of health professionals compared with the general public. AB - BACKGROUND: The study aimed to compare the beliefs of health professionals about the potential helpfulness of various mental health interventions with those of the general public. METHOD: Surveys were carried out in Australia of 872 general practitioners, 1128 psychiatrists, 454 clinical psychologists and 2031 members of the public. Respondents were presented with a case vignette describing either a person with depression or one with schizophrenia. Respondents were asked to rate the likely helpfulness of various types of professional and non-professional help and of pharmacological and non-pharmacological interventions. RESULTS: The professionals gave much high ratings than the public to the helpfulness of antidepressants for depression, and of antipsychotics and admission to a psychiatric ward for schizophrenia. Conversely, the public tended to give much more favourable ratings to vitamins and minerals and special diets for both depression and schizophrenia, and to reading self-help books for schizophrenia. CONCLUSION: The beliefs that health practitioners hold about mental disorders differ greatly from those of the general public. There is a need for mental health education campaigns to help close the gap between professional and public beliefs. PMID- 9337976 TI - Ethnicity and use of acute psychiatric beds: one-day survey in north and south Thames regions. AB - BACKGROUND: Previous studies have shown higher rates of psychiatric admissions, compulsory admissions, and diagnosed schizophrenia in Black ethnic groups compared with other population groups. METHOD: In a point-prevalence study, demographic and clinical data were collected for adult acute and low-level secure psychiatric in-patients in all National Health Service and seven private psychiatric units in North and South Thames regions on 15 June 1994. RESULTS: A total of 3710 adult acute and 268 low-level secure psychiatric patients were surveyed; 75% of the patients were White, 16% were Black, and 4% were Asian. Analysis reveals that a high proportion of the Black population were admitted to a psychiatric unit; that Black patients are more likely to be admitted under Section; to be located in locked wards; have an inpatient diagnosis of schizophrenia; and not be registered with a general practitioner. CONCLUSIONS: These findings demonstrate the over-representation of Black ethnic groups within current psychiatric provision. The complement of services to all minority ethnic groups should be examined in terms of access, appropriateness and quality of care. Racism awareness and staff training need to be incorporated into mental health service provision as a matter of priority. PMID- 9337977 TI - Disability, outcome and case-mix in acute psychiatric in-patient units. AB - BACKGROUND: Eighteen acute in-patient psychiatric units in Australia funded a syndicate to measure case-mix, disability and outcome of treatment. This syndicate included eight units in public general hospitals, five in stand-alone public psychiatric hospitals and five in private psychiatric hospitals. METHOD: Up to 100 in-patients admitted consecutively to each hospital (1359 in all) were assigned to a Diagnosis-Related Group (DRG), rated on the Health of the Nation Outcome Scales (HoNOS) and asked to complete the Medical Outcomes Trust Short Form 36 (SF36). These scales were administered again at discharge. Demographic information and length of stay were also recorded. Disability was measured by scores on the HoNOS and SF36 at admission, and outcome was assessed by the change in scores between admission and discharge. RESULTS: The public hospitals treated significantly more patients with schizophrenia and fewer with affective disorders, and their case load on admission was more disabled, on the whole, than that of the private hospitals. They achieved the same outcome or health gain as the private hospitals, but needed a shorter length of stay to do so. The addition of disability scores to DRG moderately increased the ability to predict length of stay. CONCLUSIONS: Routine outcome assessment using reliable and valid instruments is practical, and could lead to improvements in the quality of care for psychiatric patients. PMID- 9337978 TI - One hundred in-patient suicides. AB - BACKGROUND: The study aimed to define the characteristics and assess the clinical predictability and possible prevention of psychiatric in-patient suicides. METHOD: The coroner's files on all suicides in the Greater Montreal Region from 1 April 1986 to 31 March 1991 were examined. The medical records of each case of suspected in-patient suicide were then reviewed and rated for predictability and preventive measures taken. RESULTS: A total of 3079 suicides were recorded over this five-year period (mean annual rate of 16.4 per 100,000 inhabitants). Of these, 104 (3.4%) involved hospital in-patients. Nearly half (48%) of these in patient suicides occurred outside the hospital setting. The methods most frequently employed were hanging (36%) and jumping from high places (24%). Patients suffering from an affective disorder (45%) or schizophrenia (35%) comprised the majority of the sample. Suicides were significantly more predictable in general hospital psychiatric wards. Suicide prevention measures did not differ significantly across settings. CONCLUSIONS: The majority of in patient suicides were not highly predictable. For highly predictable suicides, the results underline the importance of actively treating and protecting these patients. PMID- 9337979 TI - Prospective study of clinical and social outcome of stay in small group homes for people with mental illness. AB - BACKGROUND: Small group homes operating according to the principles of supported housing have, during the past five years, become a cornerstone of the housing services for the long-term mentally ill in Copenhagen. METHOD: During a 2.5-year period, 47 long-term mentally ill persons were examined at the time of entry to a group home programme. Residents' psychopathology, social integration, mastery and quality of life were measured by structured interviews, including the Present State Examination (PSE-10); and their social functioning was recorded by interview with the staff. Forty-four of the residents were re-examined at follow up after a mean of 1.1 years using the same instruments. Data on hospitalisation were obtained through the Danish Psychiatric Case Register. RESULTS: Eighty-three per cent of the residents remained in the programme during the first year. They showed a significant improvement in subjective quality of life, PSE total score, social integration, functioning and hospitalisation index. The number of reciprocal supportive contacts in the social network increased. Lower baseline PSE total score was associated with adherence to the programme, and the improvement in quality of life during their stay was predicted by reduction in symptoms and improvement in social integration. CONCLUSIONS: A rehabilitation strategy of supplementing standard psychiatric treatment with a programme of small supportive group homes improves the quality of life, psychosocial functioning and community tenure of the long-term mentally ill. PMID- 9337980 TI - Effects of level of socio-economic development on course of non-affective psychosis. AB - BACKGROUND: This study explored the relation of level of socio-economic development to the course of non-affective psychosis, by extending the analysis of urban/rural differences in course in Chandigarh, India. METHOD: The proportion of 'best outcome' cases between urban (n = 110) and rural (n = 50) catchment areas were compared at two-year follow-up, separately for CATEGOS+ and non-S+ schizophrenia. RESULTS: The proportion of subjects with 'best outcome' ratings at the urban and rural sites, respectively, was similar for CATEGOS+ schizophrenia (29 v. 29%), but significantly different for non-S+ psychosis (26 v. 47%). CONCLUSIONS: The fact that in rural Chandigarh, psychoses have a more favourable course than in the urban area may be explained in large part by psychoses distinct from 'nuclear' schizophrenia. PMID- 9337981 TI - Ethnic differences in satisfaction with mental health services among representative people with psychosis in south London: PRiSM study 4. AB - BACKGROUND: Previous studies show that among Black Caribbeans there is a higher prevalence of schizophrenia and higher levels of both voluntary and compulsory admissions. These suggest that Black Caribbean patients may find psychiatric services less appropriate to their needs. The aim of this study was to establish the satisfaction with mental health services of representative psychosis patients in South London, especially in relation to ethnic group. METHOD: A random sample of all cases of psychotic disorder identified in the two sectors was interviewed using the Verona Service Satisfaction Schedule. Questionnaires from 50 Black Caribbean patients and 134 White patients were analysed. RESULTS: Black Caribbean patients, particularly those of second generation born in the UK, were significantly less satisfied with almost every aspect of the services that they received than either older Black Caribbean patients born in the Caribbean or White patients. Using multiple regression analysis it was found that among the younger Black Caribbean patients, unlike the other patients, the number of previous admissions was a significant predictor of dissatisfaction. CONCLUSION: Patients' ratings of satisfaction with mental health services are significantly worse for UK-born Black Caribbean than other patients with psychotic disorder in South London. PMID- 9337983 TI - Prevalence of dementia and depression among elderly people in black and ethnic minorities. AB - BACKGROUND: This study was designed to identify all elderly people of ethnic minorities living in a defined geographical area in inner-city Liverpool and to identify psychiatric morbidity and barriers to use of services. This paper reports the prevalence of dementia and depression. METHOD: A survey of the community was carried out using the Geriatric Mental State Examination, AGECAT and ethnically matched interviewers. The sampling frame consisted of Family Health Services Authority lists as a basis, with additional information from community lists, 'snow-balling' and a door-to-door survey. RESULTS: 418 people were interviewed, with a high percentage (55%) of young elderly (65-74) men. The prevalence of dementia ranged from 2 to 9% and of depression from 5 to 19%, and there were no significant differences in levels between English-speaking ethnic groups and the indigenous population. Higher levels of dementia were found among non-English-speaking groups. CONCLUSIONS: A complete enumeration of the elderly in ethnic minority groups is best achieved by using several different methods. Diagnosis of dementia may be misleading among those who do not speak the dominant language. PMID- 9337982 TI - Prevalence of spontaneous dyskinesia in schizophrenic and non-schizophrenic psychiatric patients. AB - BACKGROUND: Although movement disorders have been noted among patients never exposed to neuroleptic medications, the specificity of spontaneous dyskinesia to schizophrenia has rarely been examined. METHOD: By abstracting detailed case records, we compared the prevalence of dyskinetic movements between 94 neuroleptic-naive schizophrenic patients and 179 patients with other psychiatric disorders. RESULTS: Dyskinetic movements were more common among patients with schizophrenia than among those with all other diagnoses, and were most often noted in the body areas typically associated with tardive dyskinesia. CONCLUSIONS: Spontaneous dyskinesia appears to be relatively specific to schizophrenia and may be intrinsic to the pathophysiology of the disorder. PMID- 9337984 TI - Change in borderline symptoms one year after therapeutic community treatment for severe personality disorder. AB - BACKGROUND: The view that severe personality disorder (SPD) is untreatable derives from poor-quality studies of treatment outcome which use indirect measures of SPD pathology. This study evaluates the impact of psychotherapeutic in-patient treatment on core personality disorder symptoms. METHOD: 137 SPD patients completed the Borderline Syndrome Index (BSI) on referral and one year post-treatment ('admitted', n = 70) or one year post-referral ('non-admitted', n = 67); 22 of the non-admitted group were refused extra-contractual referral funding for their treatment. RESULTS: There was a significantly greater reduction in BSI scores in the treated than in the non-admitted group. Changes in BSI scores were significantly positively correlated with length of treatment. Assessment of the reliability and clinical significance of changes in individual subjects showed that the magnitude of this change was reliable and clinically significant in 42.9% of the admitted sample, compared with only 17.9% of the non admitted sample (18.2% of the unfunded group). CONCLUSIONS: Specialist in-patient treatment is effective in reducing core SPD psychopathology. PMID- 9337985 TI - Brain 5-HT function in obsessive-compulsive disorder. Prolactin responses to d fenfluramine. AB - BACKGROUND: Drugs that potentiate brain serotonin (5-HT) neurotransmission are effective in the treatment of obsessive-compulsive disorder (OCD), but it is unclear whether disturbances in brain 5-HT function play a role in the pathophysiology of OCD. METHOD: We studied the prolactin response to the selective 5-HT releasing agent d-fenfluramine in 14 non-depressed, drug-free OCD patients, and 14 healthy controls matched for age and gender. RESULTS: The prolactin response to d-fenfluramine was significantly increased in OCD patients compared with controls. CONCLUSIONS: The disparate results of studies of 5-HT neuroendocrine function in OCD make it unlikely that disturbances of brain 5-HT function play a central role in the pathophysiology of OCD. Increased brain 5-HT neurotransmission in non-depressed OCD subjects may represent an adaptive neurobehavioural mechanism which can be amplified to therapeutic advantage by treatment with 5-HT potentiating drugs. PMID- 9337986 TI - Personality disorder and psychopathology in Tourette's syndrome: a controlled study. AB - BACKGROUND: Some specialists associate a wide variety of psychopathologies with Tourette's syndrome (TS), while others suggest that there is no psychopathology specifically associated. Few controlled studies have been conducted to address this issue, and none has investigated personality disorder in TS. METHOD: Adults with TS and controls were evaluated using standardised psychiatric rating scales, including self-rated (STPCD) and clinician-rated (SCID-II) assessments of personality disorder, to investigate associations between personality disorder, other psychopathology and TS. RESULTS: Significantly more TS patients (25/39 (64%)) than controls (2/34 (6%)) had one or more personality disorders. TS subjects were also more likely to have more personality disorders. TS patients had significantly more depression, anxiety and obsessionality than controls. The SCID-II and STCPD were moderately well correlated. CONCLUSIONS: TS patients have a high prevalence of personality disorder and psychopathology when compared with controls. These results are the first to suggest a high level of personality disorder in a TS clinic population. PMID- 9337987 TI - Number needed to detain. PMID- 9337988 TI - Social course of schizophrenia. PMID- 9337989 TI - Use of seclusion, restraint and emergency medication. PMID- 9337990 TI - Psychopathological syndromes and familial morbid risk of psychosis. PMID- 9337991 TI - Treatment, outcome and predictors of response in elderly depressed in-patients. PMID- 9337992 TI - BSE and human prion disease. PMID- 9337993 TI - Risk assessment and clinical risk management. PMID- 9337994 TI - Serotonin: 5-HT2A receptor occupancy in vivo and response to the new antipsychotics olanzapine and sertindole. PMID- 9337995 TI - Fibronectin in bovine vitreous. An immunochemical study. AB - The vitreous body is a transparent gel essentially composed of hyaluronan, collagen and proteoglycans. These components are assembled in a three-dimensional structure that is maintained by self-aggregation of macromolecules and the interactions between these different macromolecules. We confirmed the presence of fibronectin in vitreous body using immunochemical methods and by indirect immunofluorescence on cryostat sections. We also determined its distribution in vitreous extracts as compared to those of collagen, proteoglycans and hyaluronan. Because of its high affinity to these macromolecules fibronectin in the vitreous appears to play an important role in strengthening and stabilizing the gel structure. PMID- 9337996 TI - Involvement of the dorsal paratrigeminal nucleus in visceral pain-related phenomena. AB - Cyclophosphamide is an antitumor agent that generates evolving cystitis through the release of toxic urinary by-products, mostly acrolein, that attack the bladder walls. Using c-fos expression, which permits quantitative analysis of neural activity, we demonstrated that the paratrigeminal nucleus is involved in processing the inputs that this disease generates. c-Fos staining in the paratrigeminal nucleus increases regularly reaching a plateau over the 4 h postinjection period during which the disease develops. The degree of staining is directly correlated with that of the subnucleus medialis of the nucleus of the solitary tract, which is one of the main structures that processes cystitis related inputs at the supraspinal level. PMID- 9337997 TI - Proteolytic fragments of insulin-like growth factor binding protein-3: N-terminal sequences and relationships between structure and biological activity. AB - Insulin-like growth factors (IGF-I and IGF-II) in biological fluids bind to high affinity binding proteins (IGFBP-1 to -6), which transport them and regulate their activities. Limited proteolysis of certain IGFBPs plays a major role in this regulation. IGFBP-3 is proteolysed in vivo and in several cell lines by serine proteases, including plasmin. In earlier studies we reproduced this proteolysis in vitro using recombinant human non-glycosylated IGFBP-3. Two major fragments were obtained, the larger retaining weak affinity for IGF-I and weakly inhibiting IGF I mitogenic effects. The smaller fragment, though lacking affinity for IGFs, is a potent growth inhibitor. These proteolytic fragments were isolated by HPLC and their N-terminal amino acids sequenced. Both major fragments contain the N-terminal region of the intact protein, the larger form corresponding to residues 1-160, and the smaller form, to residues 1-95. Kinetics experiments using the MG-63 osteoblast-like cell line showed that the larger peptide is generated before the smaller peptide, the latter probably being a product of secondary proteolysis of the former. Our data suggest that proteolysis of IGFBP-3 is intimately linked to its biological function. We propose a model for its action at cellular level. PMID- 9337998 TI - Male infertility associated with multiple mitochondrial DNA rearrangements. AB - Male sterility results from a number of characterized exogenous or genetic dysfunctions preventing normal differentiation into mobile spermatozoa. This may now be overcome by intra cytoplasmic sperm injection (ICSI). This practice does not require mobile, or even mature spermatozoa for in vitro fecondation. However, a functional respiratory chain, partly encoded by the mitochondrial DNA (mtDNA), is required for the mobility of the spermatozoa. We report the case of an infertile patient who wished to procreate. ICSI was proposed but he displayed multiple mtDNA deletions of possible nuclear origin in the spermatozoa and in the deltoid muscle. Even though mtDNA is maternally inherited, the possibility of a nuclear-driven mutation affecting the integrity of the mtDNA should be taken into account when ICSI is to be performed. Together with recent genetic in vitro manipulations in mammals, our data point to the importance of studying the mtDNA structure in human spermatozoa, and the potential risks of these non-natural practices for procreation. PMID- 9337999 TI - Simultaneous imaging of the surface and the submembraneous cytoskeleton in living cells by tapping mode atomic force microscopy. AB - Contact and tapping mode atomic force microscopy have been used to visualize the surface of cultured CV-1 kidney cells in aqueous medium. The height images obtained from living cells were comparable when using contact and tapping modes. In contrast, the corresponding, and simultaneously acquired, deflection images differed markedly. Whereas, as expected, deflection images enhanced the surface features in the contact mode, they revealed the presence of a filamentous network when using the tapping mode. This network became disorganized upon addition of cytochalasin, which strongly suggests that it corresponded to the submembraneous cytoskeleton. Examination of fixed cells further supported this assumption. These data show that, in addition to the structural information on the cell surface, the use of the tapping mode in liquid can also provide a good visualization of the membrane cytoskeleton. Tapping mode atomic force microscopy appears to be a promising technique for studying interactions between cell surface and subsurface structures, a critical step in many biological processes. PMID- 9338000 TI - [Functional relationship between fat mass, skinfolds, under or overweight and the "ideal theoretical weight"]. AB - Underscoring a functional relation between fat mass, skinfolds, under or overweight and the 'ideal theoretical weight', constitutes a fundamental datum in anthropology. Indeed, this relation, which differs according to age, sex, ethnic group and physical activity, provides information on the distribution of fat contained in the skinfolds, the under or overweight and the 'ideal theoretical weight'. The value of this relation lies not only in limiting the use of the corpulence index (W/H2) as an indicator of fat mass, but above all in the importance of physical activity and in the existence of a significant change in the distribution of fat according to the factors mentioned above. PMID- 9338001 TI - Why are we doing so much cancer gene therapy? Disentangling the scientific basis from the origins of gene therapy. PMID- 9338002 TI - Adeno-associated virus-mediated gene transfer into rat carotid arteries. AB - Gene transfer into the arterial wall may allow study of the role of specific genes in vascular pathophysiology and development of local gene therapies for vascular disorders. The feasibility of adeno-associated virus (AAV)-mediated gene transfer into isolated segments of normal and balloon-injured rat carotid arteries was studied using a recombinant AAV carrying CMVlacZ as a reporter gene. Approximately 10(6) and 10(7) infectious units (IU) of AAV were infused into 1 cm isolated segments of the carotid artery of 14 animals with the aid of a Silastic catheter and allowed to remain for 20 min. Animals were killed at different time points after infection and arteries stained for beta-gal activity. Microscopic examination demonstrated comparable gene transfer into medial and adventitial cells, with significantly higher efficiency of transduction in injured as compared with normal vessels. High levels of in vivo beta-gal expression persisted for at least 30 days after gene transfer. Thus, AAV is capable of transducing media and adventitia of rat carotid arteries, suggesting that it may constitute a useful vector for arterial gene transfer and gene therapy protocols. PMID- 9338003 TI - Eradication of tumor growth via biolistic transformation with allogeneic MHC genes. AB - Particle acceleration-mediated biolistics transformation has been rapidly adopted as a versatile technology for gene delivery since its original application as a physical method for delivery of foreign genes into higher plants. We have designed a versatile hand-held gene gun device for the genetic immunization of animals in vivo. The gene gun is driven by air blast and therefore does not require helium gas or vacuum for its function. In this report, we have employed this gene gun device to introduce allogeneic H-2Kb DNA directly into tumor nodules of K36 tumor-bearing AKR/J mice. Expression of the exogenous H-2Kb gene led to the regression of well-established K36 tumors. This tumor regression was correlated with the generation of potent secondary CTL responses against the K36 tumor cells following expression of the exogenous H-2Kb gene in the K36 tumor cells. Furthermore, it was observed that H-2Kb stimulator cells could stimulate the generation of both allogeneic and tumor-specific secondary CTL responses whereas mitomycin-C treated tumor cells could only stimulate tumor-specific secondary CTL responses. This approach of introducing allogeneic MHC genes directly into cutaneous tumor lesions via biolistics transformation of tumor cells in vivo can potentially be developed into an effective method for cancer gene therapy. PMID- 9338004 TI - Gene delivery by HVJ-liposome in the experimental gene therapy of murine glioma. AB - We investigated the delivery of foreign genes into mouse glioma cells in vivo using hemagglutinating virus of Japan (HVJ)-liposomes, which are coated by Sendai virus envelope protein. HVJ-liposomes, containing lacZ gene or herpes simplex virus thymidine kinase (HSVtk) gene-bearing plasmid DNA were applied in the meningeal gliomatosis (MG) mouse model system. Highly efficient delivery was observed in disseminated glioma cells, and 80% of MG mice expressing the HSVtk gene were cured by treatment with ganciclovir. These results suggest that this novel gene delivery system may be applicable for the in vivo gene therapy of human malignant glioma. PMID- 9338005 TI - Polyethylenimine (PEI) is a simple, inexpensive and effective reagent for condensing and linking plasmid DNA to adenovirus for gene delivery. AB - A simple and inexpensive method of condensing and linking plasmid DNA to carrier adenovirus particles is described. The synthetic polycation polyethylenimine is used to condense plasmid DNA into positively charged 100 nm complexes. These PEI DNA complexes are then bound to adenovirus particles through charge interactions with negative domains on the viral hexon. The resulting transfection complexes deliver plasmid DNA to cells by the adenovirus infectious route without interference from virus gene expression because psoralen-inactivated virus is employed. The PEI-DNA-adenovirus complexes display DNA delivery comparable to more sophisticated DNA virus complexes employing streptavidin/biotin linkage, but require no special reagents and are much easier to prepare. PMID- 9338007 TI - Anaerobic bacteria as a gene delivery system that is controlled by the tumor microenvironment. AB - A fundamental obstacle in gene therapy for cancer treatment is the specific delivery of an anticancer gene product to a solid tumor. Although several strategies exist to control gene expression once a vector is directly introduced into a tumor, as yet no systemic delivery system exists that specifically targets solid tumors. Nonpathogenic, obligate anaerobic bacteria of the genus Clostridium have been used experimentally as anticancer agents because of their selective growth in the hypoxic regions of solid tumors after systemic application. In this report we further describe a novel approach to cancer gene therapy in which genetically engineered clostridia are used as tumor-specific vectors for the delivery of antitumor genes. We have introduced into a strain of C. beijerinckii the gene for an E. coli nitroreductase known to activate the nontoxic prodrug CB 1954 to a toxic anticancer drug. Nitroreductase produced by these clostridia enhanced the killing of tumor cells in vitro by CB 1954, by a factor of 22. To demonstrate the specificity of this approach for tumor targeting, we intravenously injected the inactive spore form of C. beijerinckii, which upon transition to a reproductive state will express the E. coli nitroreductase gene. Nitroreductase activity was detectable in 10 of 10 tumors during the first 5 days after intravenous injection of inactive clostridial spores, indicating a rapid transition from spore to reproductive state. Tumors harboring clostridial spores which did not possess the E. coli nitroreductase gene were devoid of nitroreductase activity. Most importantly, E. coli nitroreductase protein was not found in a large survey of normal mouse tissues following intravenous injection of nitroreductase containing clostridia, strongly suggesting that obligate anaerobic bacteria such as clostridia can be utilized as highly specific gene delivery vectors for cancer therapy. PMID- 9338006 TI - Phase I study of immunotherapy of cutaneous metastases of human carcinoma using allogeneic and xenogeneic MHC DNA-liposome complexes. AB - The generation of strong tumor-specific immunity by in situ gene therapy is an attractive approach for the eradication of human cancer lesions. The objectives of this study were to examine the toxicities of employing the human HLA-A2, HLA B13 and the murine H-2K genes to generate tumor regression in patients with different cancer types via DC-Chol/DOPE cationic liposomes. The study was composed of two phaseI/II trials involving a total of 19 late-stage cancer patients. The patients were given four weekly injections of a DNA-liposome mixture directly into a cutaneous nodule. These procedures resulted in no significant clinical side-effects. The HLA-A2 gene gave the highest level of expression in situ. Although all patients treated had progressive systemic disease and eventually succumbed to their disease, strong local responses were generated in the treated nodules. Of the eight patients whose cutaneous nodules received HLA-A2 DNA, two completely regressed while four tumor nodules gave a partial local response. All but one of the patients who received HLA-A2-liposome mixtures and had a subsequent local response were either cervical or ovarian carcinoma patients. This local response, seen in a group of patients who had relapsed stage IV systemic metastatic disease and were refractory to all available therapies, demonstrates the generation of a strong local immune response following our in situ gene therapy protocol. Further studies to investigate the use of HLA-A2 DC-Chol/DOPE cationic liposomes for immunotherapy of cervical and ovarian cancers are warranted. PMID- 9338008 TI - beta-Gal enzyme activity driven by the HSV LAT promoter does not correspond to beta-gal RNA levels in mouse trigeminal ganglia. AB - beta-Galactosidase enzyme expression can be detected in only a small percentage of trigeminal ganglia (TG) neurons acutely and latently infected with herpes simplex virus (HSV), in which the lacZ reporter gene was placed down-stream of the latency associated transcript (LAT) promoter at the LAT locus. However, DNA quantification suggests that a larger percentage of cells is infected than is expressing beta-galactosidase enzyme. To investigate the mechanism involved in regulation of genes expressed from the LAT promoter in trigeminal ganglia, in situ hybridization and histochemical staining assays were employed to determine on a cell-by-cell basis beta-gal gene expression both at the RNA and protein level. Using a LAT promoter-driven beta-gal construct in HSV-1 strain HFEM, it was found that there were 89-fold more cells positive for beta-gal transcript than cells positive for beta-gal enzyme in acutely infected trigeminal ganglia and a 10-fold difference in latently infected trigeminal ganglia. Thus, there is a discordance between beta-gal mRNA and beta-gal enzyme levels in HFEM/LAT-lacZ infected cells during acute and latent infection, and the beta-gal reporter gene activity does not faithfully compare the LAT promoter activity between acute and latently infected tissue. In contrast, in situ hybridization and histochemical staining assays were performed in mice acutely infected with a virus in which 140 bp of the LAT promoter sequences flanking the TATA element were replaced by 1.8 kbp of the neurofilament promoter (HSV-1 HFEM/NF-lacZ). This construct showed a correlation between beta-gal mRNA and enzyme expression in trigeminal ganglia in acute and latent infections. These findings suggest that sequences at the 5' end of the beta-gal transcript influence translation of the beta-gal message. PMID- 9338009 TI - Adenovirus enhancement of polyethylenimine-mediated transfer of regulated genes in differentiated cells. AB - Efficient gene transfer is a prerequisite for analysing regulation of transfected promoters. We combined the DNA binding property of the cationic polymer polyethylenimine (PEI) and the potent endocytic activity of adenovirus in a PEI DNA-adenovirus complex which provided efficient plasmid delivery in differentiated cultured cells. We transfected 3T3-F442A adipocytes, C2.7 myocytes and FAO hepatoma cells with a construct containing the simian virus 40 promoter fused to the chloramphenicol acetyltransferase (CAT) gene, using a combination of PEI and 200 p.f.u. per cell of replication-deficient type 5 adenovirus. Resulting CAT activities varied according to the cell type reaching about 0.6, 8 and 38 units/mg protein for respectively 3T3-F442A, FAO and C2.7 cells. Increases in transfection efficiencies were 140- to 300-fold when compared with those obtained with PEI alone. Then we tested physiologically regulated promoters: the phosphoenolpyruvate carboxykinase gene promoter in 3T3-F442A or FAO cells and the hexokinase II gene promoter in C2.7 myocytes. Gene expression was appropriately increased by clofibrate, dexamethasone and insulin for 3T3-F442A, FAO and C2.7 cells, respectively. Thus, the combination of PEI and adenovirus is a simple, efficient, inexpensive and versatile method of gene transfer which is applicable to several differentiated cells and provides a physiologically coherent transgene regulation. We name this method PEI-adenofection. PMID- 9338010 TI - Expression of beta-galactosidase in mouse brain: utilization of a novel nonreplicative Sindbis virus vector as a neuronal gene delivery system. AB - Sindbis virus expression has been used for in vitro investigations of antigen processing, presentation and epitope mapping. The recent development of a replication-deficient recombinant Sindbis virus expression vector has made in vivo expression possible with minimal pathogenic risk. Advantages of Sindbis virus over other available viral systems include a comparatively smaller genome size making it possible to clone larger inserts, the ability to infect a wide range of host cell types with reduced pathogenicity for humans. These features suggest the possible utility of Sindbis virus for the in vivo delivery of genes to neural cells. We used the recombinant Sindbis viral expression system to target delivery of the lacZ gene to neuronal cells of mice by stereotactic surgery. Sindbis viral mRNA obtained by in vitro transcription was used to transfect baby hamster kidney (BHK) cells. As shown by histochemistry, beta galactosidase (beta-gal) was expressed in approximately 50% of transfected cells. Cells were then cotransfected with DH-BB helper sequences that enabled the recombinant Sindbis virus RNA packaging. Nonreplicative Sindbis viral stock was collected 24 h after transfection. BHK cells were then infected with viral stock and histochemistry analysis was performed. Again, approximately 50% of the cells expressed beta-gal. The same viral stock was infused into the nucleus caudatus/putamen and nucleus accumbens septi and histochemical analysis of frozen sections from the relevant brain areas confirmed that beta-gal was expressed in neurons in a time-dependent manner. beta-Gal was detected at 24 h after inoculation and was present for at least 14 days, with maximum expression at 48 h. These results suggest that a nonreplicative Sindbis virus expression system may be useful for delivery of foreign genes into the central nervous system (CNS). PMID- 9338012 TI - A CD2/CD28 chimeric receptor triggers the CD28 signaling pathway in CTLL.2 cells. AB - We are evaluating strategies to enhance the in vivo proliferation and function of adoptively transferred antigen-specific T cells. Although the CD28 costimulatory pathway is important for T cell activation and proliferation, the expression of the ligands for CD28 is highly restricted. We have generated a chimeric receptor composed of the signaling domains of CD28 and the extracellular domain of CD2 which binds the widely expressed ligand CD58. The CD2/CD28 chimeric receptor was introduced into CTLL.2 cells via retrovirus infection and was shown to be expressed on the cell surface. By monitoring early and late components of the CD28 signaling pathway, the chimeric receptor was demonstrated to trigger the CD28 pathway in response to CD2 cross-linking. The possible utility of the CD2/CD28 chimeric receptor for adoptive immunotherapy is discussed. PMID- 9338011 TI - The influence of polymer structure on the interactions of cationic polymers with DNA and morphology of the resulting complexes. AB - Four cationic polymers used to deliver DNA into cultured cells: polylysine, intact polyamidoamine dendrimer, fractured polyamidoamine dendrimer and polyethylenimine, are examined for their ability to interact with DNA. Complexes between the polymers and DNA were examined using electron microscopy. Similar toroidal structures with diameters of 55 +/- 12 nm were formed from all of the cationic polymers with DNA. The DNA complexes of the fractured dendrimer and polyethylenimine were observed as single, distinct units; their apparent diameters in solution as measured by dynamic light scattering ranged from 90 to 130 nm. The DNA complexes of polylysine and intact dendrimer generally appeared as clusters in electron micrographs; their diameters in solution were larger than 1000 nm, which suggests that their toroidal complexes aggregate in solution. The cationic polymers bind to DNA in a stoichiometry that is nearly 1:1 in primary amines to DNA phosphates. The apparent binding of all cationic polymers to DNA decreases linearly with increasing ionic strength, up to 0.8 M NaCl. Thus, at the concentrations studied, these polymers interact electrostatically with DNA forming a unit structure with toroidal morphology; the extent of aggregation of the unit structures in solution depends upon the characteristics of the individual polymer. PMID- 9338013 TI - Adenovirus-mediated gene transfer of a human IL-6 antagonist. AB - IL-6 is a pleiotropic cytokine and plays a major role in inflammation and in the immune response. Altered serum levels of IL-6 have been described in several pathologies such as myeloma, EBV-lymphoma and chronic autoimmune disease. Here we report data on the utilization of a hIL-6 receptor superantagonist with a gene therapy approach. The superantagonist used in this work possesses very high affinity for the hIL-6 receptor, and is therefore an excellent candidate for the treatment of IL-6-dependent diseases. To obtain an efficient in vivo delivery method, we constructed a recombinant adenovirus expressing the IL-6 receptor superantagonist by inserting the cDNA, controlled by the RSV promoter, into a first generation replication-incompetent adenoviral vector. Recombinant virus allowed correct expression of the transgene in vitro. Supernatants of infected cells specifically inhibited IL-6-induced transcriptional activation in hepatoma cells and blocked the IL-6-dependent proliferation of human myeloma cells. After intravenous injection of the recombinant virus into mice, nanomolar amounts of antagonist were produced in the serum, and these were able completely to inhibit IL-6 bioactivity. Gene transfer of such an antagonist offers a practical means of imposing long-term blockade of IL-6 activity in vivo for investigational and therapeutic purposes. PMID- 9338016 TI - Inhibition of HIV-1 replication by retroviral vectors expressing monomeric and multimeric hammerhead ribozymes. AB - Retroviral vectors were engineered to express monomeric and multimeric hammerhead ribozymes targeting one and nine highly conserved sites within the HIV-1 envelope (Env) coding region. In vitro, both the monomeric and multimeric ribozymes were shown to be active and cleave the target RNA containing the cleavage sites. A human CD4+ T lymphocyte-derived MT4 cell line was stably transduced with retroviral vectors expressing these ribozymes. Ribozyme expression in stably transduced cells was confirmed by Northern blot analysis and reverse transcription polymerase chain reaction (RT-PCR). As compared with the control cells lacking any ribozyme, HIV-1 replication was delayed in monomeric RzEnv expressing cells. Virus replication was almost completely inhibited in multimeric RzEnv1-9-expressing cells as no viral RNA or protein could be detected in these cells and in their culture supernatants for up to 60 days after infection. The genomic DNA from RzEnv1-9-expressing cells was shown to contain HIV-1 proviral DNA sequences at days 3 and 60 after HIV infection. HIV-1 used in the challenge experiments was found to contain fully reverse transcribed '-' strand DNA which should have been able to infect complete proviral DNA synthesis and integrate within the cellular genome without being affected by pre-existing ribozymes. Therefore, the proviral DNA present at day 3 after infection may have originated from infection by such DNA-containing virus particles. The results obtained with the retroviral vector expressing RzEnv1-9 are very encouraging and we envisage its future use in anti-HIV-1 gene therapy. PMID- 9338015 TI - Constitutive expression of murine CTLA4Ig from a recombinant adenovirus vector results in prolonged transgene expression. AB - The administration of soluble muCTLA4Ig around the time of adenovirus vector mediated gene transfer into murine hepatocytes has been shown to markedly prolong transgene expression, diminish the formation of adenovirus neutralizing antibody, decrease T cell proliferative response and infiltration into the liver without causing irreversible systemic immunosuppression. In this study, an E1/E3-deleted adenovirus vector constitutively expressing murine CTLA4Ig (Ad.RSV-muCTLA4Ig) was constructed in order to determine if production of muCTLA4Ig from within transduced cells (i.e. hepatocytes) would provide a more specific/localized interference with the CD28/B7-1 and B7-2 signaling pathways, and thus result in prolonged transgene expression in vivo at nonimmunosuppressive serum concentrations. In contrast to C3H mice receiving a control adenovirus, transduction with 6 x 10(9) p.f.u. of Ad.RSV-muCTLA4Ig and a reporter adenovirus (2 x 10(9) p.f.u. of Ad.PGK-hAAT) resulted in prolonged reporter gene expression, reduced anti-adenovirus and anti-hAAT antibody production, and attenuated T cell proliferation and IFN-gamma production in response to adenoviral vector. Mice given a constant total amount of adenovirus with diminishing amounts of Ad.RSV muCTLA4Ig and a constant amount of reporter virus (2 x 10(9) p.f.u. of Ad.PGK hAAT) demonstrated prolonged reporter gene expression and decreased anti adenovirus and anti-hAAT antibody production only when high serum levels of muCTLA4Ig were produced. Taken together, these findings suggest that a certain threshold of muCTLA4Ig must be achieved to alter the immune responses and prolong transgene expression from adenoviral vectors. PMID- 9338014 TI - NF-kappa B as a target for anti-inflammatory gene therapy: suppression of inflammatory responses in monocytic and stromal cells by stable gene transfer of I kappa B alpha cDNA. AB - One of the most challenging issues of anti-inflammatory gene therapy is the complexity of inflammatory pathways. Transcription factor NF-kappa B plays a pivotal role in activation of multiple inflammatory molecules, and therefore represents the logical target for intervention. We evaluated the feasibility of suppressing the inflammatory responses in different cell lines through specific inhibition of NF-kappa B by gene transfer of I kappa B alpha, the naturally occurring intracellular inhibitor of NF-kappa B. The I kappa B alpha overexpressing cells were established using retroviral gene transfer or stable transfection with the wild-type (wt) I kappa B alpha cDNA. In all cell types, overexpression of wt I kappa B alpha resulted in a profound (> 100-fold) increase of the I kappa B alpha message and a moderate (two- to three-fold) increase of the I kappa B alpha protein. The effects of the I kappa B alpha overexpression on the NF-kappa B activation and the inflammatory responses varied significantly in different cell lines. In conditionally immortalized human endometrial stromal cells, overexpression of I kappa B alpha prevented both interleukin-1 (IL-1) inducible degradation of endogenous I kappa B alpha protein and activation of NF kappa B. Accordingly, induction of cytokines interleukin-8 (IL-8) and Gro gamma was markedly suppressed. In monocytic THP-1 cells, both lipopolysaccharide (LPS) inducible degradation of I kappa B alpha and NF-kappa B activation were only partially inhibited by overexpression of exogenous I kappa B alpha cDNA. None the less, the LPS-induced transcription of IL-1 beta and secretion of cytokines interleukin-6 (IL-6) and IL-8 were virtually abolished. In epithelial HT-29 cells, no inflammatory responses were inhibited. These results demonstrate the range of responses in various cell lines to gene transfer of I kappa B alpha and indicate the feasibility of suppression of inflammatory responses in appropriate target cells and their progeny by suppression of NF-kappa B. PMID- 9338017 TI - Enhancement of retroviral production from packaging cell lines expressing the human immunodeficiency type 1 VPU gene. AB - The HIV-1 Vpu protein stimulates virus production by enhancing the release of viral particles from infected cells. Interestingly, Vpu was also shown to enhance the release of capsids produced by gag gene contructs of other retroviruses that lack a Vpu-like activity. To investigate the effect of Vpu expression on viral particle production in retroviral packaging cell line, we developed the Damp-VpuP cell line in which vpu expression is under the control of the tetracycline responsive promoter. Retroviral production was measured by dosage of virion associated reverse transcriptase activity, by capsid protein immuno-detection in cell-free supernatants and by evaluating the transfer of antibiotic resistance to target cells. Induction of the Damp-VpuP cell line caused a 40-fold increase in the titer of infectious virus-like particles when compared with control cell lines. This increase in viral titer was not the result of a clonal effect nor was it a consequence of high selective pressure but rather the effect of a Vpu mediated enhancement of viral particle production. Similar results using the third generation psi CRIP packaging cell line confirmed these findings. Constitutive expression of vpu caused a 13-fold increase in viral titer in this packaging cell line. These results indicate that the expression of HIV-1 vpu in retroviral packaging cell lines can significantly improve the titers of infectious retroviral particles. PMID- 9338018 TI - RT-PCR method specific for the detection of transgenic CFTR mRNA in the presence of transgene plasmid DNA and endogenous CFTR mRNA. AB - A major problem with experiments involving the correction of cystic fibrosis by gene transfer has been reliably detecting the transgenic message and distinguishing this from an endogenous message and from vector DNA. We have exploited the SV40 small T antigen intron present in an expression vector containing the CFTR cDNA (pCFAS) using a primer directed to the spliced sequence to allow specific and precise detection of the transgeneic message. PMID- 9338019 TI - Adverse drug reaction of the month--a new series. PMID- 9338020 TI - Maintenance pharmacological immunosuppressive strategies in renal transplantation. AB - Current maintenance immunosuppressive regimens for transplantation are based on three classes of drugs: corticosteroids, immunophilin-binding agents (eg, cyclosporin and tacrolimus), and antimetabolites (eg, azathioprine and mycophenolate). Drugs from the various classes inhibit the immune system at different points and are thus synergistic when used in combination. PMID- 9338022 TI - Handbooks, learning contracts, and senior house officers: a collaborative enterprise. AB - Efforts to bring about improvements in hospital-based education and training for senior house officers over the last few years have raised issues which are gradually being addressed. One of these is the lack of understanding by many clinical teachers of educational principles and their application to senior house officer training. This study describes how volunteer consultants in five specialties in a North West District General Hospital worked together to develop an educational structure for senior house officers. An audit of education and training was carried out across the hospital to help identify problem areas. An education specialist worked with consultants to develop, implement and evaluate a handbook based on adult learning principles. The handbook incorporated a learning contract, formal review process and a curriculum of learning objectives for each specialty. In parallel, consultants created in-house videos which were used to raise awareness of clinical teachers in the hospital about these educational issues. Preliminary evaluation showed positive responses by both senior house officers and consultants to both the study and its outcomes. PMID- 9338021 TI - Langerhans cell histiocytosis (histiocytosis X). AB - There has been a renewed interest in Langerhans cell histiocytosis in recent years due both to advances in basic research and to improvements in diagnostic and treatment approaches. In this article, we review the various aspects of the disease and the potential implications of these recent scientific researches for our understanding and management of the disorder. PMID- 9338023 TI - Octreotide scanning for carcinoid tumours. AB - The somatostatin analogue octreotide may be used in the diagnosis of carcinoid and other neuroendocrine tumours. Radionuclide scanning following intravenous injection of 111Indium-labelled octreotide (111In-DTPA-pentetreotide) provides a sensitive, non-invasive method of localising somatostatin-positive tumours. The technique may also be used to identify patients who may respond to 'cold' octreotide therapy and to monitor therapeutic efficacy. PMID- 9338025 TI - The use of angiotensin-converting enzyme inhibitors in the treatment of heart failure in hospital practice. AB - Several well-controlled trials in patients with heart failure have shown that the use of angiotensin-converting enzyme (ACE) inhibitors, in combination with a diuretic, causes a reduction in mortality and morbidity, which seems to be mainly due to a reduction in fatal and nonfatal cardiovascular events. Our aim was to determine whether 249 consecutive patients discharged from hospital with a primary diagnosis of heart failure were routinely being treated with an ACE inhibitor at an appropriate dose. At the time of admission to hospital, 91 (36.5%) were receiving a combination of a diuretic and an ACE inhibitor, 129 (51.8%) were receiving a diuretic alone, and 29 (11.6%) had not previously received either a diuretic or an ACE inhibitor. At the time of discharge from hospital all patients were on a diuretic and 144 (57.8%) were also receiving an ACE inhibitor. Although 41 patients (16.5%) had a relative or absolute contraindication for the use of an ACE inhibitor, 64 patients (25.7%) with no contraindication were not receiving an ACE inhibitor. Many of the patients who were prescribed an ACE inhibitor were given it at an inappropriate dose; only 24 patients (16.7%) were on the dose that was used in the clinical trials showing a reduction in mortality. These results show that in one in four patients admitted to hospital with heart failure who should be receiving an ACE inhibitor by the time of discharge, are not. The average age of these patients was 76 years. Whilst it has been shown that the benefit of ACE inhibitors does not appear to be age-related, most published studies have not included many patients over the age of 80. Specific studies looking at the effect of ACE inhibitors in elderly patients would be helpful, as well as studies to determine the optimum treatment regimen for this age group. PMID- 9338024 TI - Primary infection by type 1 human immunodeficiency virus: diagnosis and prognosis. AB - Primary infection by type 1 human immunodeficiency virus (HIV) is symptomatic in about 70% of cases. The acute illness is a mononucleosis-like syndrome with characteristics such as mucosal ulcerations. The duration and severity of the symptoms appear to be related to the prognosis. After reviewing the most frequent signs and symptoms of primary HIV infection, we report different prognostic studies which examined the association between the acute illness and the progression of HIV disease. PMID- 9338026 TI - The role of ultrasound in the diagnosis of a large, rapidly growing, thyroid mass. AB - The value of ultrasound in the diagnosis of a large rapidly growing thyroid mass was assessed in a study of 42 patients with a large (> 3 cm) rapidly growing (< two months) solitary mass. Haemorrhage into a thyroid nodule was present in 31 patients and thyroid malignancy in 11. Ultrasound of haemorrhage into a thyroid nodule revealed a large cystic mass in all 31 patients containing internal debris (22), septations (three), or a combination of both (six). The malignant causes of a large rapidly growing mass were lymphoma (two), anaplastic carcinoma (four) and metastasis (five). Ultrasound of these thyroid malignancies revealed a mass with a smooth, well-defined margin and strikingly low homogeneous echogenicity in all cases. Patients with thyroid metastases had evidence of widespread metastatic disease elsewhere. Lymphoma was differentiated from anaplastic carcinoma on fine needle aspiration cytology or surgical biopsy. Ultrasound was of value in differentiating between a benign haemorrhagic nodule and a malignant tumour. The various malignant tumours had similar appearances, however, and could not be distinguished on ultrasound. PMID- 9338027 TI - Permanent cardiac pacing in elderly patients with recurrent falls, dizziness and syncope, and a hypersensitive cardioinhibitory reflex. AB - The study was designed to assess the outcome of treatment with permanent dual chamber pacing of elderly patients with falls, dizziness and syncope associated with the demonstration of a hypersensitive cardioinhibitory reflex. Questionnaires were sent to patients (and their general practitioners) who had been referred to a regional pacing centre with recurrent falls, dizziness or syncope diagnosed as likely to be secondary to cardioinhibitory carotid sinus syndrome or predominantly cardioinhibitory vasovagal syndrome. After pacemaker insertion, 84% of patients had no further syncope over a mean follow-up period of 10 (range 1.5 to 30) months. Minor symptoms persisted in only 40% of all patients. Symptoms were unchanged in 22%. It was concluded that permanent dual chamber pacing is an effective treatment for elderly patients with recurrent falls, dizziness and syncope in whom a hypersensitive cardioinhibitory reflex is found. Good results were obtained in this group with a simple diagnostic work-up. PMID- 9338028 TI - Cranial and peripheral neuropathy due to Epstein-Barr virus infection. AB - A case of Epstein-Barr virus infection with neurological complications is described. An 18-year-old man developed cranial neuropathy and peripheral sensorimotor polyneuropathy three weeks after a sore throat. Though severely affected initially, he made a good recovery and no specific therapeutic intervention appears to have influenced his clinical course. PMID- 9338029 TI - Complete gastric outlet obstruction following acid ingestion complicated by acute pancreatitis and disseminated intravascular coagulation. AB - A case is described where accidental acid ingestion resulted in the development of oesophageal stricture and complete gastric outlet obstruction. Following a smoothly conducted pre-surgery endoscopic examination the patient developed acute pancreatitis, which, on initial clinical examination, was diagnosed as a viscus perforation. The severity of pancreatitis was such as to lead to disseminated intravascular coagulopathy and ultimately death. The interest lies in the fact that pancreatitis was precipitated in a previously traumatised stomach by such an innocuous procedure as fibre-optic endoscopy. PMID- 9338030 TI - Paraplegia due to thoracic disc herniation. AB - Disc herniation at the thoracic the spine level is more common than generally thought. Localisation of pain may be vague and may erroneously point to cardiopulmonary, gastrointestinal, genito-urinary or even psychiatric disease. Magnetic resonance imaging is the investigation of choice, especially if spinal cord compression is suspected. PMID- 9338031 TI - Diagnostic difficulties in periodic Cushing's syndrome. AB - A 22-year-old black woman presented with symptoms suggestive of Cushing's syndrome three years after chemotherapy for a presumed teratoma with cervical lymphadenopathy. Initially, the absence of clinical signs and the demonstration of two normal 24 h urinary free cortisols appeared to exclude the diagnosis, but an ectopic adrenocorticotropin-producing thymic carcinoid was subsequently removed surgically. Cushing's syndrome due to ectopic adrenocorticotropin production can be difficult to diagnose, particularly if there is periodic hormonogenesis. PMID- 9338032 TI - Male breast tuberculosis. AB - Tuberculosis of the breast is rare and tuberculosis of the male breast is not a recognised entity. We describe a man with tuberculosis of the breast which was clinically thought to be a malignancy. PMID- 9338033 TI - Tumour seeding following percutaneous endoscopic gastrostomy placement in head and neck cancer. AB - We report the case of a patient with advanced squamous carcinoma of the supraglottic larynx and hypopharynx who developed metastatic gastric deposits occurring at the site of a percutaneous endoscopic gastrostomy tube, inserted 10 months previously by the pull technique. We review seven previous reports of tumour deposits occurring at the site of placement of a percutaneous endoscopic gastrostomy in patients with head and neck cancer, and consider alternative methods of enteral feeding in such patients. PMID- 9338034 TI - Hospital-acquired hyperkalaemia. PMID- 9338036 TI - A treatable cause of lymphocytic meningo-encephalitis. PMID- 9338035 TI - Photosensitivity and lymphoma. PMID- 9338037 TI - Potentiation of the anticoagulant effect of warfarin. PMID- 9338038 TI - Cranial neuropathy. PMID- 9338039 TI - Acute hyponatraemia. PMID- 9338040 TI - Paralysis after treatment for thyrotoxicosis. PMID- 9338041 TI - Ophthalmic complications of HIV/AIDS. PMID- 9338042 TI - Cannabis and alcohol in stroke. PMID- 9338043 TI - Occult CO poisoning presenting as an epileptic fit. PMID- 9338044 TI - Report of a workshop on problem-based learning and its implications for medical education in the UK. Held 7 June 1996 at the Royal Society of Medicine, London. PMID- 9338045 TI - Hepatitis B vaccine and neurotoxicity. PMID- 9338046 TI - Reduced forced expiratory flow in schoolchildren with respiratory symptoms: the Odense Schoolchild Study. AB - In the present population-based study, spirometric lung function was assessed in symptomatic schoolchildren with and without asthma as compared to an asymptomatic reference group. The primary aim was to investigate if impaired lung function could be demonstrated in symptomatic schoolchildren, even in the absence of diagnosed asthma. Spirometry [forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), 50% of forced expiratory flow (FEF 50%) and 75% of forced expiratory flow (FEF 75%)] and anthropometric measures (standing height, weight, skin fold thickness, and length and circumference of the upper arm) were obtained from 1369 8-10-year-old children (81.5% of the eligible population) during the school year 1985-86. In 1321 of those subjects (96.5% of those examined), a self administered questionnaire was completed. Thirty-five children belonging to ethnic minorities were excluded, thus 1286 subjects were included for further analysis. Point prevalences concerning asthma and respiratory symptoms (wheeze, cough and shortness of breath) were obtained. Thirty-seven children reported asthma and one or more asthma-like symptoms (symptomatic asthmatics), whereas 40 children denied having asthma, although claiming one or more asthma-like symptoms (symptomatic non-asthmatics). In both symptomatic groups, FEF 50% and FEF 75% were reduced relative to the reference group, the deficit being larger in the symptomatic asthmatics. FEF 75% was found to be more reduced than FEF 50%. FEV1 and FVC did not differ significantly between groups. It is concluded that only half of the schoolchildren with respiratory symptoms usually associated with the presence of asthma actually reported having this disease. These results demonstrate the presence of reduced lung function in symptomatic, reportedly non asthmatic, children, suggesting clinically important underdiagnosis of asthma. More severe impairment of lung function was found in known asthmatics, also implying some degree of undertreatment. PMID- 9338047 TI - The long-term efficacy and safety of two different corticosteroids in chronic sarcoidosis. AB - Deflazacort (DFZ) is claimed to have fewer adverse bone effects than prednisone (PDN) at doses with equivalent anti-inflammatory activity (5 mg PDN = 6 mg DFZ). However, its safety over the long-term has never been tested in a controlled trial. The aim of the present study was to assess prospectively the safety and efficacy of DFZ compared with PDN in previously untreated patients with chronic, histologically proven sarcoidosis needing long-term (> or = 2 yr) corticosteroid therapy. Thirty-six patients were treated with PDN for 32 +/- 18 months and 36 patients were treated with DFZ for 42 +/- 18 months, and followed-up with periodic chest X-ray, 67Gallium lung scan, angiotensin converting enzyme (ACE), serum and urinary calcium levels, spirometry, alveolar diffusion (DLCO) arterial oxygen tension (PaO2), bone mineral content (BMC) (by computed tomography), and a complete biochemical and haematological profile. The two groups were similar as regards sex, age, pulmonary and extrapulmonary involvement, parameters of activity and impairment, and initial BMC. Daily starting doses were 23.2 +/- 11.4 mg DFZ and 22.3 +/- 6.9 mg PDN. One year of trial was completed by 69 patients, 2 yr by 59 patients, 3 yr by 46 patients and 4 yr by 24 patients. Some patients were followed-up for 5-7 yr. The mean daily dose over the whole period was 15 +/- 10 mg DFZ and 10 +/- 6 PDN, starting from 21 +/- 9 and 15 +/- 8 mg in the first year, and progressively declining to a mean of 9 +/- 6 mg in both groups in the fourth year. Chest X-ray, 67Ga score, ACE and forced vital capacity improved significantly in both groups. Urine total calcium improved significantly in the PDN group (345 +/- 27 to 186 +/- 47; P < 0.05) with a similar but non-significant pattern in the DFZ group (270 +/- 28 to 207 +/- 39). Non-significant improvements were observed in DLCO, PaO2 and forced expiratory volume in 1 s in both groups. Drug-related adverse events were more frequent in the PDN group, causing discontinuation of the drug in four PDN patients. Body weight increased mainly in the PDN group [69.9 +/- 0.4 to 73.6 +/- 0.8 kg vs 70.1 +/- 0.4 to 70.0 +/- 0.6 kg in the DFZ group (P < 0.01)]. Bone mineral content dropped under the fracture threshold in most PDN patients, who thus appeared at higher risk for fractures. In fact, six atraumatic skeletal fractures were observed in this group but only one in the DFZ group. Two further patients in the DFZ group and eight in the PDN group were obliged to start corrective measures for bone loss and/or bone pain. At the end of the study, 21 patients (12 DFZ, nine PDN) no longer needed corticosteroids, and the others were taking a maintenance daily dose that controlled the disease adequately. In conclusion, DFZ appeared as effective as PDN in the long-term treatment of chronic sarcoidosis, and it may have fewer side effects, especially on bone. PMID- 9338048 TI - The prevalence of self-reported asthma and respiratory symptoms in Ankara, Turkey. AB - The prevalence of self-reported asthma was studied in a group of Turkish adults using the European Community Respiratory Health Survey (ECRHS) questionnaire distributed during 1994 local elections in Ankara, Turkey. A total of 2020 questionnaires were issued and 1820(90%) were returned. The mean age of the subjects was 34.5 +/- 10.2 years. The prevalence of wheezing at any time in the past was 39.1% which is much higher than has been reported in the literature. However, only 21.7% of the study population had wheezing in the year preceding the survey and 2.9% of them had severe asthma attacks. The prevlaences of nocturnal wheeze, nocturnal cough and morning tightness were higher in females (P = 0.05 for each). The results of this study showed a high rate of reported symptoms but a low rate of diagnosis and treatment of asthma among the adult population in Ankara. PMID- 9338049 TI - Low IgG subclass levels in brittle asthma and in patients with exacerbations of asthma associated with respiratory infection. AB - Serum total immunoglobulin (G, A, M and E) and IgG subclass levels were studied in 23 patients with brittle asthma, 23 age- and sex-matched patients with mild asthma and 33 patients with recurrent infective exacerbations of their asthma. Patients with brittle asthma showed significantly reduced levels of IgG (mean +/- SD, 8.8 +/- 3.3 g l-1) compared to patients with mild asthma (11.0 +/- 2.5 g l-1) (P < 0.008) with further significant reductions in the brittle compared to the mild group in IgG1 (5.2 vs 6.3, P = 0.035), IgG2 (2.4 vs 3.25, P < 0.006), IgG3 (0.39 vs 0.55, P < 0.05) and IgA (1.91 vs 2.38, P < 0.03). There were no significant differences between the brittle group and the group with recurrent infective exacerbations for any parameter, but the latter group showed significantly reduced levels of IgG (8.2, P < 0.001), IgG1 (4.9, P < 0.00001) and IgG2 (2.5, P < 0.02) compared to the mild group. In all groups, there was no relationship between dose of inhaled steroids and levels of any antibody. These findings suggest that the presence of a mild degree of humoral immunodeficiency relates to severity of asthma, and suggests that immunoglobulin replacement therapy may be appropriate in patients with the more severe forms of asthma. PMID- 9338050 TI - The diagnostic value of cold air hyperventilation in adults with suspected asthma. AB - The diagnostic value of isocapnic hyperventilation of cold air (IHCA) is not fully established. All 342 adult patients in whom IHCA had been performed because of a clinical suspicion of asthma between 1992 and 1994 were analysed retrospectively in the authors' hospital. In addition, 26 healthy subjects were recruited. According to strict criteria, the patients were divided into asthmatics and symptomatic non-asthmatics. For the calculations of sensitivity, specificity and accuracy, the symptomatic non-asthmatic group served as a control. The post-test probability of asthma after IHCA was determined for all the possible pre-test probabilities by applying Bayes' theorem. A linear regression model was used to investigate the factors associated with the reactivity to IHCA. A single 4-min IHCA and skin prick tests were performed in the healthy subjects. Of the 287 patients in the final analysis, 113 were defined as asthmatics and 174 as symptomatic non-asthmatics. The accuracy was highest using a 9.0% fall in forced expiratory volume in 1 s (FEV1) as a cut-off value; the specificity was then 86.8% and the sensitivity 31.9%. The authors found IHCA to be a useful diagnostic test only if the pre-test probability of asthma is between 0.30 and 0.56. The positive final diagnostic gain of IHCA is 22% at its best, but the negative gain is negligible for all possible pre-test probabilities. Factors associated with reactivity to IHCA were young age and, to a lesser extent, a history of cold-weather-associated respiratory symptoms and pre-challenge bronchial obstruction. If a rigid cut-off value for a positive response is used in all age groups, the specificity of IHCA is good but the sensitivity is unacceptably low in adults. The diagnostic value of IHCA might increase if age is taken into account when defining the cut-off value. PMID- 9338051 TI - The single-breath nitrogen test in coal miners: factors associated with failure to perform. AB - The single-breath nitrogen washout (SBN2) test was used, along with spirometry, in the baseline examination of a longitudinal study in a cohort of active coal miners from North-eastern France. The procedure was computerized, allowing the technician to coach and encourage the subject, and excluding computation errors. While all miners performed satisfactory spirometry, a significant number were unable to meet the National Heart and Lung Institute recommendation concerning a 10% agreement of vital capacities. When the limits were set at +/-12%, 57 miners (24.2%) were still classified as failing to perform. When compared to those who succeeded, those failing proved to be significantly older, had more cumulated dust exposure, a higher prevalence of chronic cough and sputum, and a trend for more micronodulation on the chest radiographs. The ventilatory function did not differ between the two groups. These results confirm previous data on spirometric test failure concerning older age and respiratory symptoms, extending them to the SBN2 test. The present study further indicates that dust exposure and roentgenologic pneumoconiosis nodulation are associated with failure to perform the SBN2 test. PMID- 9338052 TI - Genetic relatedness of Burkholderia (Pseudomonas) cepacia isolates from five cystic fibrosis centers in Michigan. AB - Burkholderia cepacia isolates from patients with cystic fibrosis (CF) attending five CF centers were studied for relatedness by cellular fatty acid methyl esters (FAME) and by chromosomal DNA restriction analysis. Twenty-eight of 32 (87.5%) isolates tested were grouped in cluster group 1 based on their FAME profiles. DNA analysis revealed that 29 of 32 (90.6%) B. cepacia isolates from five CF centers had one closely related DNA pattern. To examine strain variation over a time period, FAME profiles and DNA patterns of isolates from serial cultures on seven patients from center D were studied. For four patients, all serial B. cepacia isolates belonged to a single FAME cluster group; for the remaining three patients, all serial isolates belonged to any two of the four cluster groups. On serial culture isolates, a single DNA pattern (pattern A) was found in 31 of 32 isolates demonstrating a close genetic relatedness. These data corroborate the observations that the majority of patients colonised with B. cepacia in a CF center harbor strains genetically closely related as determined by FAME profiles and DNA patterns. PMID- 9338053 TI - Multiple pulmonary nodules in association with pyoderma gangrenosum. AB - This report describes a patient with extensive pyoderma gangrenosum in whom there were co-existent lung abnormalities. The patient's X-ray showed peripherally sited multiple pulmonary lesions bilaterally. A lung biopsy showed chronic non specific inflammatory changes with neutrophil and lymphocyte infiltration which were similar to the skin lesions. This case was diagnosed as multiple aseptic nodules in pyoderma gangrenosum. The pulmonary infiltrative shadows were controlled only with prednisolone treatment. Steroid therapy is considered to be the first choice to control pulmonary lesions of this disease. PMID- 9338054 TI - Tracheobronchopathia osteochondroplastica presenting at the time of a difficult intubation. AB - Tracheobronchopathia osteochondroplastica (TO) is a rare and usually benign disorder affecting the trachea and occasionally the bronchi. We describe the case of a 46-year-old woman who was discovered to have TO at the time of a difficult intubation. This case was also unusual since the patient had presented no previous symptoms despite the presence of extensive endotracheal and bronchial lesions. The incidence of TO appears to be underestimated in the literature in view of the fact that it is usually benign. However, a more accurate estimate of its true prevalence may become available through the use of bronchoscopy and computerized tomographic scanning. PMID- 9338055 TI - Tracheobronchopathia osteochondroplastica. AB - A case of tracheobronchopathia osteochondroplastica (TO) was diagnosed in a 68 year-old male with prolonged cough. A bronchoscopy revealed multiple nodular excrescences along the anterolateral wall of the trachea and main bronchi. Tissue specimens showed pronounced change of bronchial cartilage with massive mineralization diagnostic for TO. The literature on the subject is reviewed here. The aetiology and pathogenesis is unknown. The severity of TO range from no symptoms to severe dyspnoea, haemoptysis or pneumonitis. Treatment is seldom necessary. However, in severe cases, bronchoscopic removal of obstructing excrescences and surgery has been performed with therapeutic effect. Differential diagnosis of nodular excrescences includes amyloidosis, endobronchial sarcoidosis, calcificating lesions of tuberculosis, papilomatosis and tracheobronchial calcinosis. Awareness of the condition as a differential diagnosis to neo-plasms is important, to avoid unnecessary surgery or chemotherapy. PMID- 9338056 TI - Multiple tracheal strictures following mechanical ventilation. AB - Patients presenting with features of airway narrowing (cough, wheeze, exertional breathlessness and obstructive spirometry) may be suffering from either localized or generalized airway obstruction. Doctors sometimes overlook the possibility of localized obstruction (whether due to tumour, foreign body aspiration or stenosis), and patients may experience symptoms for a long time before the correct diagnosis is made. PMID- 9338072 TI - Foulds' dangerous idea revisited: the multistep development of tumors 40 years later. PMID- 9338073 TI - Cancer cells exhibit a mutator phenotype. AB - This review analyzes the concept and evidence in support of a mutator phenotype in human cancer. The large number of mutations reported in tumor cells cannot be accounted for by the low mutation rates observed in normal somatic cells; rather, it must be a manifestation of a mutator phenotype present early during the tumorigenic process. The interaction between increased mutagenesis and clonal selection provides a mechanism for the selection of cells with increased proliferative advantage. The concept of a mutator phenotype in cancer has gained considerable support from the findings of enormous numbers of somatic mutations in repetitive sequences in human tumors. Moreover, cell lines exhibiting microsatellite instability demonstrate an increased mutation frequency in expressed genes. A knowledge of mechanisms that generate multiple mutations in cancer cells has important implications for prevention. For many tumors, a delay in the rate of accumulation of mutations by a factor of two could drastically reduce the death rates from these tumors. PMID- 9338074 TI - Increasing complexity of Ras signal transduction: involvement of Rho family proteins. PMID- 9338075 TI - B-Myb: a key regulator of the cell cycle. PMID- 9338076 TI - Alterations in DNA methylation: a fundamental aspect of neoplasia. AB - Neoplastic cells simultaneously harbor widespread genomic hypomethylation, more regional areas of hypermethylation, and increased DNA-methyltransferase (DNA MTase) activity. Each component of this "methylation imbalance" may fundamentally contribute to tumor progression. The precise role of the hypomethylation is unclear, but this change may well be involved in the widespread chromosomal alterations in tumor cells. A main target of the regional hypermethylation are normally unmethylated CpG islands located in gene promoter regions. This hypermethylation correlates with transcriptional repression that can serve as an alternative to coding region mutations for inactivation of tumor suppressor genes, including p16, p15, VHL, and E-cad. Each gene can be partially reactivated by demethylation, and the selective advantage for loss of gene function is identical to that seen for loss by classic mutations. How abnormal methylation, in general, and hypermethylation, in particular, evolve during tumorigenesis are just beginning to be defined. Normally, unmethylated CpG islands appear protected from dense methylation affecting immediate flanking regions. In neoplastic cells, this protection is lost, possibly by chronic exposure to increased DNA-MTase activity and/or disruption of local protective mechanisms. Hypermethylation of some genes appears to occur only after onset of neoplastic evolution, whereas others, including the estrogen receptor, become hypermethylated in normal cells during aging. This latter change may predispose to neoplasia because tumors frequently are hypermethylated for these same genes. A model is proposed wherein tumor progression results from episodic clonal expansion of heterogeneous cell populations driven by continuous interaction between these methylation abnormalities and classic genetic changes. PMID- 9338077 TI - Ara-C: cellular and molecular pharmacology. AB - The antimetabolite cytosine arabinoside (ara-C) represents a prototype of the nucleoside analog class of antineoplastic agents and remains one of the most effective drugs used in the treatment of acute leukemia as well as other hematopoietic malignancies. The ability of ara-C to kill neoplastic cells is regulated at three distinct but interrelated levels. First, the activity of ara-C depends on conversion to its lethal triphosphate derivative, ara-CTP, a process that is influenced by multiple factors, including nucleoside transport, phosphorylation, deamination, and levels of competing metabolites, particularly dCTP. Second, the antiproliferative and lethal effects of ara-C are linked to the ability of ara-CTP to interfere with one or more DNA polymerases as well as the degree to which it is incorporated into elongating DNA strands, leading to DNA fragmentation and chain termination. Finally, the fate of the cell is ultimately determined by whether a threshold level of ara-C-mediated DNA damage is exceeded, thereby inducing apoptosis, or programmed cell death. The latter process is influenced by components of various signal transduction pathways (e.g., PKC) and expression of oncogenes (e.g., bcl-2, c-Jun), perturbations in which may significantly alter ara-C sensitivity. A better understanding of these factors could eventually lead to the development of novel therapeutic strategies capable of overcoming ara-C resistance and improving therapeutic efficacy. PMID- 9338078 TI - Protein misassembly in vitro. PMID- 9338079 TI - Oligomer formation by 3D domain swapping: a model for protein assembly and misassembly. PMID- 9338080 TI - The structure of amyloid fibrils by electron microscopy and X-ray diffraction. PMID- 9338081 TI - Transthyretin quaternary and tertiary structural changes facilitate misassembly into amyloid. AB - Human transthyretin (TTR) can be transformed into amyloid fibrils by partial acid denaturation to yield a monomeric amyloidogenic intermediate that self-associates into amyloid through quaternary structural intermediates, which are identified by sedimentation velocity methods. The monomeric amyloidogenic intermediate has substantial beta-sheet structure with a nonnative but intact tertiary structure as discerned from spectroscopic methods. Proteolysis sensitivity studies suggest that the C-strand-loop-D-strand portion of TTR becomes disordered and moves away from the core of the beta-sandwich fold upon formation of the monomeric amyloidogenic intermediate over the pH range 5.1-3.9. The single site mutations that are associated with early onset amyloid disease [familial amyloid polyneuropathy (FAP)] function by destabilizing tetrameric TTR. Under mild denaturing conditions, the FAP variants populate the monomeric amyloidogenic intermediate conformation, which assembles into amyloid, whereas wild-type TTR remains tetrameric and nonamyloidogenic. The FAP mutations do not significantly alter the native folded structure; instead, they appear to act by making the thermodynamics and perhaps the kinetics more favorable for formation of the amyloidogenic intermediate. Suppressor mutations have also been characterized that strongly stabilize tetrameric TTR and disfavor the formation of the monomeric amyloidogenic intermediate, thus inhibiting amyloid formation. The mechanistic details characterizing transthyretin amyloid fibril formation available from the biophysical studies outlined within have been utilized to develop a new therapeutic strategy for intervention in human amyloid disease. This approach features small molecules that bind with high affinity to the normal fold of transthyretin, inhibiting the quaternary and tertiary structural changes associated with the formation of the monomeric amyloidogenic intermediate that self-assembles into amyloid. Ligand binding to TTR stabilizes the native tetrameric fold, which is nonamyloidogenic. PMID- 9338082 TI - Domain stability in immunoglobulin light chain deposition disorders. PMID- 9338083 TI - Mutational effects on inclusion body formation. PMID- 9338084 TI - Times to exhaustion at 90, 100 and 105% of velocity at VO2 max (maximal aerobic speed) and critical speed in elite long-distance runners. AB - Previous studies had concluded that the treadmill velocity-endurance time hyperbolic relationship for runs could be accuratly approached with a regression at condition that bouts of exercise duration were included between 2 and 12 min. This regression allows to calculate the critical speed (CS) defined as the slope of the regression of work (distance) on time to exhaustion, the anaerobic running capacity (ARC) being the intercept of this line (Monod & Scherrer, 1965). The purpose of this investigation was to give practical indication concerning the choice of the velocities in reference to the maximal aerobic speed (MAS i.e. the minimum speed which elicits VO2max). Subjects were fourteen elite male long distance runners (27 +/- 3 years old; VO2max = 74.9 +/- 2.9 ml.kg-1.min-1, MAS = 22.4 +/- 0.8 km.h-1, CS = 19.3 +/- 0.7 km.h-1 and 86.2 +/- 1.5% MAS). tlim 100 values (321 +/- 83 s) were negatively correlated with MAS (r = -0.538, p < 0.05) and with CS (km.h-1) (r = -0.644, p < 0.01). tlim 90 (1015 +/- 266 s) was positively correlated with CS when expressed in % MAS (r = 0.645, p < 0.01) and not when expressed in km.h-1 (r = -0.095, P > 0.05). tlim 105 (176 +/- 40 s) only was correlated with ARC (r = 0.526, p < 0.05). These data demonstrate that running time to exhaustion at 100 and 105% of MAS in a homogeneous elite male long-distance runners group is inversely related to MAS. Moreover, tlim 90 is positively correlated with CS (%MAS) but neither with tlim 100 and 105 nor with maximal aerobic speed. So from a practical point of view, the velocities chosen to determine the critical speed, would be closed to the maximal aerobic speed (time to exhaustion around 6 min), taking into account that the tlim 105 is correlated with the anaerobic capacity, whereas tlim 90 is correlated with the critical speed. PMID- 9338085 TI - Interleukin-2 activates a sub-population of cutaneous C-fibre polymodal nociceptors in the rat hairy skin. AB - We have investigated the effects of interleukin-2 on identified cutaneous C- and A delta- fibre nociceptors in an in vivo rat saphenous nerve preparation. A fraction of C-polymodal (33%), A delta- (22%) and C- (7.5%) mechanical nociceptors were activated by intradermal injection of interleukin-2. For C-fibre polymodal nociceptors, concentration thresholds were < or = 0.12 unit/3 microliters and the percentage of interleukin-2-activated nociceptors did not increase with concentrations above 0.06 unit/3 microliters. Responses were dose dependent and characterized by potent tachyphylaxis for subsequent injections of the same dose. C-fibre polymodal nociceptors activated by interleukin-2 were also activated by subsequent chemical stimuli as follows: 81% were activated by histamine (300 ng/3 microliters), 87% by bradykinin (75 ng/3 microliters), 100% by topical acetic acid and 87% by capsaicin (3 micrograms/3 microliters). In contrast, C-fibre polymodal nociceptors that could not be activated by interleukin-2 responded less frequently to histamine (17%) and bradykinin (24%) but were generally activated by noxious chemicals, including acetic acid (82%) and capsaicin (70%). In conclusion, this study demonstrates that interleukin-2 is a potent activator of a discrete population of cutaneous C-polymodal nociceptors, which are chemosensitive to endogenous inflammatory mediators. The fact that these nociceptors respond to a variety of endogenous mediators is consistent with the concept of multiple humoral mechanisms of itch and, consequently, the difficulties in reducing itch associated with inflammation. PMID- 9338086 TI - Metabolic inhibition and chloride transport in isolated toad skin. AB - 1. When added to the Na(+)-containing solution bathing the isolated toad skin, dinitrophenol (DNP, an uncoupler of oxidative phosphorilation) caused decreases in the baseline values of short circuit current (SCC) and transepithelial conductance (G). 2. DNP also inhibited the increases in SCC and G caused by theophylline, whether added prior to the xanthine, or after the effect of the latter was fully developed. 3. In skins exposed to theophylline and bathed in Cl( )-free (sulfate Ringer's) solution, the changes in SCC and G had a similar time course (t1/2 > 15 min). In the presence of Cl- (skins bathed in Ringer's solution), SCC decreased with a similar rate, whereas the rate of the decrease in G was greater (t1/2 < 15 min). 4. DNP also decreased the SCC induced by a Cl- concentration gradient in skins exposed to theophylline (SCCg) with a time course similar to its effect on the theophylline-increased G in the presence of Cl-. DNP was effective irrespective of the presence of ambient Na+. 5. A similar difference was observed in skins bathed in CIR and exposed to forskolin. In contrast to theophylline, however, forskolin partially overcame the inhibition of G brought about by DNP; no such recovery was observed in SCC. 6. In contrast to its influence on the responses to theophylline and forskolin, DNP failed to prevent either the increase in G or the onset of SCCg in skins exposed to dibutyryl cyclic AMP. 7. Rotenone, an inhibitor of the electron-transport chain, significantly decreased SCC and G in the unstimulated skin. It also prevented the SCC response to theophylline, and decreased it if added after the effects of the xanthine were fully developed, but failed to modify the increase in G brought about by theophylline. The time course of SCC inhibition by rotenone was similar to that caused by DNP. 8. Ouabain, an inhibitor of Na+,K(+)-ATPase, decreased SCC in the theophylline-stimulated skin, without affecting G. 9. We conclude that, whereas integrity of oxidative energy metabolism is necessary to sustain SCC in the isolated toad skin, it is not a strict requirement for the increase of Cl(-) dependent G activated by cAMP. 10. The effect of DNP on Cl(-)-dependent G activated by cAMP is probably exerted at the cAMP generation step, by inhibition of adenyl cyclase and/or a decrease in the availability of ATP. PMID- 9338087 TI - Glutamate-arginine salts and hormonal responses to exercise. AB - Hormonal changes during exercise is of growing interest because of their role in adaptation, and performance. The production of amino acids (AA) due to the degradation of muscle protein increases during exercise and some AA may be utilized for energy expenditure or as hormonal secretagogues. Thus, one can propose a strategy to reduce muscle protein breakdown and regulate hormones involved in energy metabolism by dietary AA supplementation. We assessed the effects of glutamate-arginine salt (AGs) ingestion on exercise-induced hormonal alterations in highly trained cyclists (age 18-22 yrs). Using an indwelling catheter, we collected multiple blood samples at rest, during warm up, during and after an intense exercise session. Plasma growth hormone (hGH), insulin and cortisol were measured by radioimmunoassay. As reported in previous studies, we observed a marked increase in plasma hGH and cortisol levels during and after exercise in the placebo (Pl) condition as well as a slight decrease in insulin concentration. In addition, we found that the ingestion of AGs had significant effects on some dynamic hormonal changes. AGs had no effect on resting plasma levels of hGH, insulin or cortisol. However, the marked elevation in cortisol and hGH during and after exercise in the placebo condition, was greatly diminished when subjects ingested AGs. Our results show that AGs can modify exercise-induced hormonal changes and raise the possibility that it may be used to alter energy metabolism during endurance exercise. PMID- 9338088 TI - Severe modifications of ether- ester- or diester-linked glycerolipid and non glycerolipid synthesis in human neuroblastoma LAN-1 cells cultured with octadecylmethylglycerophosphocholine. AB - The growth rate of the human neuroblastoma LAN-1 cells was decreased half after 48 h of incubation with dialkylglycerophosphocholine 1-O-octadecyl 2-O-methyl-sn glycero-3-phosphocholine (ET-18-O-CH3), at 4 microM. Four radiolabelled precursors, [3H]hexadecanol, [3H]hexadecanoic, [3H]arachidonic acids, or N acetyl[14C] neuraminic acid were added in the culture medium to follow their cell incorporation among various glycerolipids, gangliosides and eicosanoids. Several modifications of the glycerophospholipid synthesis induced by ET-18-O-CH3 were observed. The inhibition of 1-O-[3H]hexadecanoyl 2-O-acyl glycerophosphocholine synthesis provoked the accumulation of diacylglycerophosphoethanolamine. The inhibition of 1-O-[3H]hexadecyl 2-O-acyl glycerophosphocholine and ethanolamine synthesis enhanced the synthesis of non phosphorous glycerolipids with 1-O [3H]hexadecyl-sn-glycerol backbone. The eicosanoid synthesis was not disturbed. Alterations of ether- ester- and diester-linked glycerophospholipid and non phosphorous glycerolipid synthesis could modify the lipid membrane distribution and affect the enzymatic pathway of the ceramide synthesis. An excess of [3H]hexadecanoyl-amide molecular species of ceramides in mono- and disialo gangliosides was observed. By contrast the N-acetyl [14C]neuraminosyl-linkage in these two groups of gangliosides never reached the hypersialylation process described during sialoglyco-peptide synthesis in murine carcinoma kidney cell lines. Both metabolic disturbances of the glycerolipid and ganglioside synthesis reported for the neuroblastoma LAN-1 cell line sensitive to ET-18-O-CH3 extend and confirm the previous studies with other more resistant cell lines from the murine Meth A sarcoma and the rat colon carcinoma. PMID- 9338089 TI - Treadmill chamber for studies of respiratory gas exchange in the rat during exercise. AB - A treadmill for studying gas exchange in small mammals during exercise is presented. The system consists of a motor-driven running mat enclosed in a gastight chamber that receives a measured flow of air from a compressed air cylinder. The gas flow and temperature, pressure and instantaneous gas composition of the chamber (oxygen, carbon dioxide and water) are measured continuously and the data are computed to include the effects on chamber atmosphere of the rat activity, either running or at rest. The system is completed with a shock delivery grid that stimulates the rat to run. The calculations are based on the changes in the composition of the gas in the chamber (constantly stirred by a small electric fan) induced by the rat instead of relying on the alterations induced in the outflowing gas. The consumption of oxygen, and production of carbon dioxide and water by the rat are computed in real time, giving a very fast response to physiological change induced by exercise. The chamber is custom-made from an aluminium block and a plexiglass lid; all other components are available commercially. The system, as described, allows for a detailed analysis of respiratory gas (and water) exchange by rats under varying exercise conditions, there is practically no time lag between changes in respiratory gases and the detection of these changes, and the buffering effect of the chamber size is practically eliminated because of the calculation approach used. PMID- 9338090 TI - Enhanced luminescence study of liver homogenate response to oxidative stress. AB - An enhanced luminescence technique was used to monitor the response of liver homogenates stressed with sodium perborate. Rat liver homogenates were subjected to oxidative stress with sodium perborate, and the light signals, generated by a suitable system, containing luminol and compounds producing enhancement of light emission such as sodium benzoate and indophenol, were detected by a luminometer. The intensity of light emission (E) was found dependent on homogenate concentration (C). When C increased, E at first increased as well and, then, decreased rapidly. The graphic expression of this phenomenon resulted as a curve that can be described by the equation: E = a.C/exp(b.C). It is proposed that the a value represents the capacity of the tissue to catalyze the production of .OH radical species. The b value might be related to the capacity of the tissue to scavenge such radicals, since it increases when homogenates are supplemented with antioxidants and decreases when homogenates are treated with prooxidant. The results obtained by supplementing homogenates with iron containing substances, or using model systems, suggest that cell substances catalyzing the luminescent reaction, such as the hemoproteins, are "scavengers" as well as radical producers. The concentration-emission curve obtained with suitable model system is described by the equation: E = a.C/exp(b.Ck). It is suggested that, using the k value, information can be obtained on the relative capacity of hemoproteins and antioxidant systems to interact with .OH radicals. PMID- 9338091 TI - Effects of compression rate on rats carotid blood flow. AB - The effects of compression rate on carotid blood flow were investigated in awake rats submitted to hyperbaric experiments conducted up to 70 bar (7 MPa, gauge pressure) with either slow compression (0.1 bar/min), i.e., inducing only mild High Pressure Neurological Syndrome (HPNS), or fast compression (2 bar/min), i.e., in the earlier time course of experiments leading to epileptic seizures. Implanted transit-time ultrasonic flowprobes were used, and data were analyzed by regression methods. Mean carotid blood flow increased with ambient pressure during either low or high compression rate, but the increase was significantly more important with the latter. These results evidenced that carotid blood flow increased with ambient pressure, and moreover that this enhancement depends on compression rate. PMID- 9338092 TI - Isoproterenol-induced subcellular redistribution of G-protein beta subunits in S49 lymphoma cells demonstrated by a novel competitive ELISA. AB - A novel competitive ELISA has been developed for the determination of levels of the beta subunit of guanine-nucleotide-binding protein (G-protein) using antipeptide antibodies directed against the amino terminus of the beta subunit. Because beta subunits form highly hydrophobic.heterodimeric complexes with gamma subunits of G-proteins, specific assay conditions were required. Optimal concentrations of antibodies, detergents, Mg2+ as well as ionic strength were determined. In addition, we found that an effective binding of the used antibodies to the beta subunit was ensured only after denaturation of the beta gamma complexes. Subsequently, this ELISA was used for quantitation of the beta subunit in subcellular fractions of S49 lymphoma cells during isoproterenol mediated desensitization of beta-adrenergic controlled transmembrane signalling system. A 10 min as well as 60 min treatment of the cells with isoproterenol (1 nmol/ml) resulted in a significant shift of G-protein beta subunits (presumably as beta gamma complexes) from the plasma membrane fractions to low-density microsomal fractions. No significant change was detected after the hormone action in the distribution of plasma membrane constitutive enzymes. In conclusion, the developed ELISA helped us to reveal that beta-adrenergic stimulation can induce redistribution of the beta gamma dimer from plasma membranes to low-density microsomes. PMID- 9338093 TI - Checkpoint controls and cancer. Introduction. PMID- 9338094 TI - Control of the G1/S transition. AB - On the basis of current knowledge, control of the G1/S phase transition is largely a matter of regulating a set of specific cyclin dependent kinase (CDK) activities. In mammalian cells, the G1/S specific CDK activities are composed of complexes between D type cyclins and either CDK4 or CDK6 and between cyclin E (and possibly cyclin A) and CDK2. A variety of internal and external signals regulate G1/S specific CDKs by modulating cyclin availability, the levels of CDK inhibitory proteins and the phosphorylation status of CDKs. Although much is now known about the regulation of G1/S specific CDKs, the only well characterized substrate to date is the retinoblastoma gene product, RB. Phosphorylation of RB by CDKs neutralizes its cell cycle inhibitory properties, allowing progression of G1 to S phase. Not surprisingly, many components of the cell cycle regulatory machinery, including CDKs, CDK inhibitors and CDK substrates, are important targets of mutations that lead to human malignancy. PMID- 9338095 TI - S phase damage sensing checkpoints in mammalian cells. AB - Mammalian cells have evolved multiple responses for dealing with DNA damage. One response is to acutely downregulate DNA synthesis at the initiation step. Essentially nothing is known about the initial signal that activates this SDS pathway or the macromolecules involved in transducing the signal into the final inhibitory step at origins. Determining whether any radiation induced changes in known proteins involved in cell cycle regulation or in other signal transduction pathways are primary or secondary responses to DNA damage constitutes a major challenge to identifying members of the pathway. It may turn out to be easier to identify the final mediator in the pathway, namely the protein(s) whose interaction with origins is ultimately affected by radiation. Hopefully, mutations in SDS genes in genetically tractable systems such as S cerevisiae or Schizosaccharomyces pombe will allow the identification of homologous genes in mammals. Most tumour cells are TP53 negative, and yet it is not clear that TP53 status influences radiation sensitivity. The SDS pathway may therefore represent an important protective mechanism that stands in the way of effective tumour cell killing by radiation therapy. It is hoped that an understanding of this pathway will provide opportunities for developing novel antineoplastic targets and/or radiation sensitizers. PMID- 9338096 TI - Cyclins and the G2/M transition. AB - The entry of a cell into mitosis is regulated by an elaborate network of kinases and phosphatases that control both for the timing of cell division and the complete reorganization of the cellular architecture. The mitotic cyclin/Cdks form part of large multiprotein complexes whose other components are only now beginning to be identified. The continuing identification of proteins that contribute to these complexes and changes in the composition of these complexes are likely to give a more integrated view of how mitotic cyclin/Cdk complexes are regulated and how they function-not only to induce mitosis, but also to aid further mitotic progression. Furthermore, assigning specific G2/M functions to distinct mitotic cyclin/Cdk complexes will require the identification of differences in substrate specificities between the mitotic cyclin/Cdk complexes, perhaps in parallel with specific cyclin knockouts in mice. Such investigations will be complicated by potential functional overlap between mitotic cyclin/Cdk complexes in vitro and in vivo. Although cyclin/Cdk1 is thought to be the major kinase that initiates the onset of mitosis, a more complete understanding of how cells move from G2 to a mitotic state will require further identification of kinases operating upstream, downstream and in parallel with Cdk1, their substrates and their relationship with one another during the G2/M transition. PMID- 9338097 TI - The DNA replication licensing system. AB - The Xenopus cell free system has proved a good model system to study in vitro DNA replication and the mechanism preventing rereplication in a single cell cycle. Studies using this system resulted in the development of a model postulating the existence of a replication licensing factor (RLF), which binds to the chromatin before the G1-S transition of the cell cycle and is displaced during replication. The nuclear envelope prevents rebinding of RLF and hence relicensing. Nuclear envelope breakdown at mitosis is required to allow another round of replication. Protein kinase inhibitors block licensing factor activity and arrest Xenopus extracts in a G2 like state. These kinase inhibitors have allowed the development of an in vitro assay leading to the biochemical purification of RLF components. RLF can be separated into RLF-B and RLF-M, the latter consisting of several members of the MCM/P1 class of replication proteins. In Xenopus as well as in many other eukaryotes, the binding of MCM/P1 proteins to chromatin before S phase is essential for replication to occur. The proteins are then displaced as replication proceeds. These changes in subnuclear distribution are reflected by changes in the phosphorylation status. MCM/P1 proteins do not bind to the DNA on their own but need RLF-B to be loaded onto the chromatin. Their cycling behaviour is reminiscent of the existence of a prereplicative complex at the origins of replication in yeast, suggesting that the licensing mechanism is ubiquitous in eukaryotes. PMID- 9338098 TI - Cyclin dependent kinase inhibitors. AB - Progression through the eukaryotic cell cycle is regulated by the activities of a family of cyclin dependent kinases (CDKs). These kinases are negatively regulated by phosphorylation and by the action of cyclin kinase inhibitors (CKIs). In mammalian cells, two classes of CKIs have been identified, the INK4 class and the CIP/KIP class. These CKIs are versatile negative regulators of CDK function and have potential roles in development, checkpoint control and tumour suppression. Analysis of CKI knockout indicates that although these inhibitors are not generally required for survival, the phenotypes observed span the gamut of what might be expected for loss of a cell cycle inhibitor. This chapter summarizes our current understanding of the roles of CKIs in growth control. PMID- 9338099 TI - Yeast checkpoint controls and relevance to cancer. AB - Checkpoint controls arrest cells with defects in DNA replication or DNA damage. For several reasons, checkpoint controls may be relevant to ontogeny and treatment of cancer. Firstly, mutations in two human genes, TP53 and ATM, give rise to cellular defects in cell cycle checkpoints and are associated with cancer. Secondly, although checkpoint defects potentially render the cell damage sensitive, they may do so only in combination with other defects in the cell's response to damage. Therefore, manipulation of checkpoint defects, requiring a description of normal and mutant pathways, will be required for this type of therapeutic approach. Those pathways are being described in yeast cells. In budding yeast, the study of checkpoint genes has led to the view that these genes have many roles in the cellular responses to DNA damage, including roles in arrest in multiple stages of cell cycle, in transcriptional induction of repair genes, in DNA repair itself and additionally some undefined role in DNA replication. The checkpoint pathways and proteins that carry out these responses may consist of sensor proteins that detect damage, signaller proteins that transduce an inhibitory signal and target proteins that are altered to arrest cell division (or cause other changes in cell behaviour). Yeast genes that may act at each step have been identified, leading to a working model of checkpoint pathways. An initial step in the pathway may involve the processing of damage to an intermediate that signals arrest and acts in DNA repair. Human checkpoint pathways may have defects in processing damage as well. PMID- 9338100 TI - The anaphase promoting complex. AB - We have proposed a preliminary model of how the anaphase promoting complex functions throughout the cell cycle, but despite the flurry of recent publications characterizing the APC--its components, regulation and substrate specificity--many fundamental questions remain to be answered. Firstly, the remaining components of the APC need to be identified and characterized. We do not know if all cyclosome components are conserved in all eukaryotes, or if higher eukaryotes, having a more complicated cell cycle machinery, maintain additional subunits for more sophisticated functional and regulatory control. In addition, we need to determine the identity of the various kinases and phosphatases that regulate the APC itself. The biochemistry of individual APC components is also a mystery, and a specific biochemical function has not been assigned to any known members of the complex. It is not at all clear which subunit(s) of the complex actually recognizes the E2 enzyme and which subunit(s) recognizes the cyclin destruction box. It is likely that many cyclosome substrates remain to be identified, and it will be interesting to determine whether all cyclosome substrates require a destruction box for their degradation or whether the APC recognizes other determinants of protein instability. Finally, we assume that the APC degrades mitotic cyclins in all proliferating cells, but whether it degrades unique cell cycle related substrates in specific tissues is unclear. Furthermore, nothing is known about APC function during meiosis, or whether the APC degrades other substrates that are not related to the cell cycle. This is an exciting and rapidly developing field in the exciting world of cell cycle biology. We expect that new findings will surely reveal many interesting surprises about this essential protein complex. PMID- 9338101 TI - Mammalian G1 and G2 phase checkpoints. AB - This present review explores the mechanisms for DNA damage induced G1 and G2 arrest in mammalian cells. The complexity of the TP53 pathway is attested to by the variety of genes regulated by TP53, many of which require further investigation to bring their importance into focus. One gene intensely studied, p21, has been linked to the G1 arrest mechanism and may, like TP53, be involved in some aspect of DNA repair. The outcome of TP53 activation for cell survival is equally complex and relies much upon cellular context and the type of DNA damaging agent employed. Although TP53 may participate in sensing DNA damage, additional components are likely to be required. Much of the focus on defining the mechanism of G2 arrest in mammalian cells has concentrated on the cyclin B1/CDC2 kinase. Activation of this kinase is suppressed by DNA damage, and this may result from the imposition of inhibitory phosphorylations on the CDC2 kinase as well as downregulation of cyclin B1 levels. The logical point where the G2 checkpoint interacts with the CDC2-CDC25C autocatalytic loop to prevent CDC2 activation remains to be defined and could involve inhibition of CDC25C-CDC2 interaction. It is hoped that moving upstream of CDC2 towards the point where DNA damage is sensed by the cell will uncover homologues of yeast components implicated in G2 checkpoint control. The finding that certain G2 checkpoint abrogators preferentially synergize with DNA damaging agents in cells with defective TP53 provides a potential pharmacological route through which TP53 defective cells might be targeted for destruction. Further exploration of this vulnerability might prove useful for future anti-cancer drug discovery efforts. PMID- 9338102 TI - Maintaining genetic stability through TP53 mediated checkpoint control. AB - TP53 serves as a key relay for signals elicited by cellular stresses arising from diverse environmental or therapeutic insults. This relay then activates a cell cycle arrest or cell death program, depending on the stimulus and cell type. The absence of TP53 function disables the cell death or arrest programmes, thereby allowing the emergence of variants with various types of genomic alterations. The data discussed focus on two different types of signals that trigger the TP53 relay system. Firstly, TP53 arrests cell cycle progression in response to the types of DNA damage most commonly detected in cells undergoing tumour progression. Secondly, TP53 is activated by specific depletion of ribonucleotide pools, which prevent cells from entering S phase under conditions that could lead to chromosome breakage. The contribution of both responses limits the emergence of genetic variants. The DNA damage induced arrest appears to be triggered by as few as one double strand break in normal human fibroblasts. Analysis of the arrest kinetics after ionizing radiation shows that TP53 activates a prolonged arrest response in cells with irreparable DNA damage and that high efficiency cell elimination is achieved by a process that can be activated over multiple cell cycles. These data indicate that the primary function of the TP53 arrest/apoptosis pathway in response to double strand break is to eliminate damaged cells from the proliferating population, not to allow additional time for lesion repair. However, it remains possible that repair of other types of damage may benefit from TP53 mediated arrest. Analyses in model genetic systems indicate that the absence of TP53 function allows, but does not ensure, a high intrinsic rate of genetic variation and that instability is increased substantially when cells proceed through S phase under inappropriate growth conditions. This implies that inactivation of TP53 function in combination with other genetic alterations, such as oncogene activation, could accelerate genomic instability and tumour progression. PMID- 9338103 TI - Functions of the DNA dependent protein kinase. AB - The DNA dependent protein kinase (DNA-PK) is a trimeric nuclear complex consisting of a large protein kinase and the Ku heterodimer that regulates kinase activity by its association with DNA. Recent findings have shown structural similarities between DNA-PK and a family of lipid and putative protein kinases (PIK family). DNA-PK is one of the PIK members known to be a protein kinase with clearly identified effector subunits. A broad range of observations link DNA-PK to dual roles in double strand DNA break (DSB) repair and transcription. Unlike its most closely related PIKs, DNA-PK is not required for activating cell cycle regulated DNA damage signalling mechanisms. Instead, the phenotypes and biochemical properties of DNA-PK are most consistent with functions in DSB repair and joining steps in recombination mechanisms. DNA-PK is associated with RNA polymerase II and RNA polymerase I transcription complexes, where it most frequently has a negative regulatory role. PMID- 9338104 TI - Telomerase, checkpoints and cancer. AB - Telomere dynamics and changes in telomerase activity are consistent elements of cellular alterations associated with changes in proliferative state. In particular, the highly specific correlations and early causal relationships between telomere loss in the absence of telomerase activity and replicative senescence or crisis, on the one hand, and telomerase reactivation and cell immortality, on the other, point to a new and important paradigm in the complementary fields of ageing and cancer. Although the signalling pathways between telomeres and transcriptional and cell cycle machinery remain undefined, recently described homologies between telomeric proteins and lipid/protein kinase activities important in chromosome stability provide evidence for the existence of pathways transducing signals originating in chromosome structure to cell cycle regulatory processes. Similarities between cell cycle arrest at senescence and the response of mortal cells to DNA/oxidative damage suggest overlap in the signal transduction mechanisms culminating in irreversible and stable cell cycle arrest. The feasibility of targeting telomeres/telomerase as a strategy for antiproliferative therapeutics has been shown in studies in yeast, in which mutations in specific telomere associated genes result in delayed cell death. Similarly, antisense oligonucleotide inhibition of telomerase activity in human tumour cells (HeLa) results in delayed cell death. The mechanism of cell death and possible escape from this fate require further study. In human cells, however, it would seem reasonable to predict that in these circumstances, apoptosis is induced in the vast majority of cells either directly in response to a DNA damage signal arising from critically shortened telomeres or as a secondary consequence of genetic instability. PMID- 9338105 TI - The ATM gene and protein: possible roles in genome surveillance, checkpoint controls and cellular defence against oxidative stress. AB - The autosomal recessive disorder ataxia-telangiectasia (AT) is highly pleiotropic. It is characterized by gradual loss of Purkinje cells in the cerebellum, leading to progressive neuromotor deterioration, immunodeficiency, developmental defects in specific tissues, profound predisposition to malignancy and acute sensitivity to ionizing radiation. AT cells show chromosomal instability, premature senesence, radiosensitivity and defects in cell cycle checkpoints activated by ionizing radiation. Several radiation induced pathways that regulate the cell cycle seem to be defective in AT cells, at least one of which is mediated by TP53. Extensive characterization of the cellular defects of AT cells, together with the recent isolation of the ATM gene, has provided some insight into the possible physiological roles of the ATM protein. Several lines of evidence, including the nature of the agents that elicit the hypersensitivity of AT cells, point to the possibility of a defect in the response to damage induced by oxidative stress, which affects various cellular macromolecules. The ATM protein might have a role in activating defence mechanisms against oxidative stress. This hypothesis broadens the previous concept of the AT defect and explains several aspects of the AT phenotype that cannot be accounted for by defective processing of DNA damage. PMID- 9338106 TI - Apoptosis and cancer mechanisms. AB - For nearly two decades, studies in the cancer research field focussed on identifying genes that act as positive and negative regulators of cell growth. Only relatively recently was it recognized that the regulation of cell death (apoptosis) is also an important modulator of tumorigenesis. At least two genes linked to human cancers, BCL2 and TP53, have been shown to regulate apoptosis. The correlation between apoptosis modulating genes and human tumours raises an important question as to how dysregulation of apoptosis contributes to neoplastic transformation and malignant cell growth. Cell culture studies have clearly demonstrated that TP53 can induce and BCL2 can suppress apoptosis in response to various stimuli. Studies of mammalian viruses, which possess mechanisms for both inducing and evading apoptosis, have also extended our understanding of this process. On the basis of such findings, several animal models have been developed which begin to address the role of apoptosis regulation in tumorigenesis. This chapter discusses those animal models, focussing on bcl-2 (and its relatives) and p53. PMID- 9338107 TI - Genetic instability in animal tumorigenesis models. AB - In this review I have attempted to a describe some of the recent mouse tumour models and their impact on our understanding of cancer aetiology. The focus has been on cell cycle regulatory genes and DNA repair genes which are likely to affect cancer development at least in part through genetic instability mechanisms. The cell cycle regulatory genes classified as tumour suppressors, TP53 and RB, maintain genomic stability and inhibit cancer through their roles in preserving cell cycle checkpoints. The cell cycle inhibitors have variable effects on cancer prevention, and their role in preserving genetic stability remains largely unexplored. The DNA repair gene models described here show the most direct connection between genetic instability and cancer, even in the absence of demonstrable cell cycle effects. It should be clear that the development of mice deficient in cell cycle control or DNA repair will provide useful tools for studying the interplay of these processes with genetic instability and cancer. Important new insights into the mechanisms of cancer initiation and progression are likely to come increasingly from such models in the coming years. PMID- 9338108 TI - Treatment-refractory depression: definitions and characteristics. AB - Forty to 50% of depressed patients who are initially prescribed antidepressant medications or administered electroconvulsive therapy do not experience a timely remission. This group typifies treatment-refractory depression (TRD), defined as a failure to demonstrate an "adequate" response to an "adequate" treatment trial (i.e., sufficient intensity of treatment for sufficient duration). The approach to the patient with TRD must be methodical. The clinician should examine potential factors contributing to apparent non-response: trial adequacy, compliance, differential diagnosis, and treatable comorbid conditions. After addressing these variables, a patient who does not demonstrate a remission may be considered treatment resistant (relative or absolute). Although many of these patients will respond to a subsequent treatment regimen, there are no (or only nominally useful) predictors for the initial selection of that "subsequent" antidepressant treatment. Hence, the initial treatment is typically chosen on the basis of safety and convenience, not differential efficacy. The search for the clinical and biological correlates of long-term or acute outcome presents a major nosologic conundrum: Who will respond to treatment? Which treatment? In this manner, TRD challenges the prognostic utility of our current phenomenologic-based diagnostic system. PMID- 9338109 TI - Treatment of refractory depression. AB - Treatment of refractory depression is a common challenge for mental health professionals. To assure optimal treatment, the clinician should first determine that adequate dosage has been employed for an adequate duration. Next there are a variety of augmentation strategies or alternative antidepressants that could be considered. This report reviews the evidence supporting these strategies, describes their implementation, compares their effectiveness, and offers suggestions for approaching the array of treatments available. PMID- 9338110 TI - Alternative approaches to refractory depression in bipolar illness. AB - Thus, there appears to be a large variety of approaches to refractory bipolar depression. In contrast to several decades ago, wherein augmentation of lithium with antidepressants and neuroleptics was essentially the only treatment mode available, a panoply of treatment options now exist. However, their relative efficacy in different illness subtypes and stages remains to be better delineated, as do their optimal sequencing and use in combination in individual patients. It is the opinion of these authors and many of our colleagues in the field that initial use of several mood stabilizer drugs in combination may have a preferable long-term outcome in some rapid cycling patients, compared with the immediate use of a unimodal antidepressant with an inadequate single mood stabilizer, although this remains to be systematically studied. The use of thyroid augmentation strategies would appear to have merit in relationship to not only the potential treatment of lithium-related hypothyroidism, but also in augmenting antimanic and antidepressant effects. As one moves toward some of the complex combination treatment strategies discussed in this chapter, one has to be particularly careful about drug interactions and their potential for toxicity as well as therapeutic effects. Perhaps a prevailing guideline would be to use these agents more carefully in combination therapy than in monotherapy, with slow upward titration of dose to individual patients' side effects thresholds, even in preference to targeting of conventional blood level windows. In this way, side effects can be avoided during the assessment of complex combination regimens. In addition, one should be aware of potential pharmacokinetic interactions. For example, with the addition of valproate to carbamazenine, one should reduce the dose of carbamazepine, as valproate will not only increase the free fraction of carbamazepine based on displacement of protein binding, but will lead to increased accumulation of carbamazepine-10,11-epoxide. This epoxide is not measured in conventional assays but could contribute to the side effects profile (Ketter and Post, 1994). Similarly, valproate will markedly increase blood levels of lamotrigine; the starting dose of this agent should be substantially lower than conventional dosage when these two drugs are used in combination. We suggest the utility of detailed mapping with a formal system-such as the Life Chart Methodology (LCM) (Leverich and Post, 1996)-of mood fluctuation vs. medications in order to optimize and rationalize complex combination therapy. In this way, not only can the nuances of partial response be better defined, but also basic decisions about the therapeutic index and relative likelihood of response can be more readily assessed. We have discussed the other merits of the life chart method as an important clinical treatment tool as well as a research tool in other venues, but reemphasize its potential great importance in the treatment of refractory cyclic bipolar patients, in whom an initial period of remission of depression may, in many instances, be as likely attributable to the natural course of illness as the current intervention being offered. As such, it behooves the clinician to have a systematic database for the more subtle issues of dose titration and sequential addition of medications in complex combination regimens. In the face of inefficiency to one combination strategy, how one moves to the next strategy remains a highly individualized, clinically-based algorithm. We suggest the potential utility of moving towards a new set of mood stabilizers and then repeating some of the unimodal antidepressant additions and augmentation trials in an attempt to overcome refractory depression. Refractory depression in bipolar patients should be viewed as a medical emergency in light of the high potential for suicide in the illness in general (Chen and Dilsaver, 1996) and in patients who have either sustained or episodic refracto PMID- 9338111 TI - Psychotherapy of refractory depressions. AB - Psychotherapy is a potentially valuable intervention for treatment-resistant depression. This review provides a brief, general overview of the use of psychotherapy to treat depression and more focused consideration of time-limited interpersonal, behavioral, and cognitive behavioral strategies for patients who are not responsive to antidepressant medication. Effective strategies emphasize individualized assessment, psychoeducation, a high level of structure and therapist activity, operationalized short-term goals, self-help and homework activities, and an empirical-collaborative approach to treatment. Although some treatment-resistant patients respond to therapy alone, more promising evidence is emerging from studies of combined strategies. PMID- 9338112 TI - Suicide: risk factors and prevention in refractory major depression. AB - The literature regarding risk factors for suicidal behavior in the context of major depressive episode is reviewed. An organized framework for prevention strategies is provided. Risk factors for suicide in major depression can be organized according to whether their effect is on the threshold for suicidal acts or whether they serve mainly as triggers or precipitants of suicidal acts. For patients sufficiently depressed to present for evaluation and treatment, severity of depression is a poor guide to risk for suicidal acts. The best predictors relate to the threshold or predisposition to suicidal acts in response to major depression. Although available literature on suicidal behavior focuses on major depression in general, the findings may be usefully applied in refractory depression. Guidelines for assessment and management of suicidal behavior in major depression are offered. PMID- 9338113 TI - Refractory depression in children and adolescents. AB - Refractory or treatments resistant depression in child and adolescent populations is a difficult construct to operationalize currently. To date, only one of the small number of completed double-blind placebo-controlled treatment investigations have not demonstrated a significant effect of antidepressants in comparison to placebo. However, it has been established that child and adolescent MDD is a serious disorder that appears to have clinical continuity with adult affective disorders and is generally of long duration with high rates of recurrence and eventual progression to mania, substance abuse, or other serious psychopathology. In addition, families of children with affective disorders evidence substantial genetic loading with high rates of affective disorders contributing both genetic vulnerability and potential environmental risk as well. There have been no empirically identified treatments that alter the long-term course of the illness. Thus treatment resistance is a significant issue for this population. This review will focus on controlled treatment trials and will examine the potential relevance of psychosocial impairment, genetic-familial risk, and neuromorphometric brain differences to treatment resistance in children and adolescents with major depression. PMID- 9338115 TI - Differences of alkaline phosphatase and arginase activities in human colorectal carcinoma cell lines. AB - Remarkable differences in the levels of alkaline phosphatase and arginase were found in colorectal cell lines tested. In HT-29 cells, which are extremely sensitive to the induction of cell differentiation, very low levels of arginase were detected. On the other hand, high levels of arginase were present in cell lines derived from highly malignant tumours. Both findings support a prognostic significance of arginase activity in colorectal carcinomas. PMID- 9338114 TI - Assignment of the cellular retinol binding protein 1 (Rbp1) and hepatic lipase (Lipc) genes to rat chromosome 8. AB - Genes for the cellular retinol binding protein 1 (Rbp1) and hepatic lipase (Lipc) were assigned to chromosome 8 of the rat by the PCR analysis of somatic cell hybrids. The current findings extend homologies between rat chromosome 8, mouse chromosome 9, and human chromosomes 3q and 15q. PMID- 9338116 TI - The influence of serotonin on follicular cells FRTL-5 in vitro. AB - Serotonin is an aromatic monoamine found in parafollicular cells of rats, bats and in a human medullary thyroid carcinoma cell line. We have examined the hypothesis that serotonin stimulates follicular cells. The effect of serotonin in presence and absence of thyrotropin was studied in a cell line of rat follicular cells FRTL-5 (Fischer Rat Thyroid cells in Low serum) by measuring [3H]thymidine incorporation into cell DNA (growth assay) and by transmission electron microscopy. The cell line was used to avoid the contamination with serotonin secreted from parafollicular cells. Results show that serotonin at 3 microM, 10 microM, 30 microM, and 100 microM concentrations increases [3H]thymidine incorporation into cell DNA. Serotonin at a 1000 microM concentration reduces sharply the [3H]thymidine incorporation into cell DNA. All cytoplasmic organelles completely disappear, only a thin fibrous membrane remains. The toxic effect of serotonin is observed in the nuclei, too. As expected, thyrotropin stimulates follicular cells. Serotonin does not have any influence on that stimulative effect of thyrotropin. Rough endoplasmic reticulum consists of round vesicles, several mitochondria are present in the cytoplasm. From the surface few pseudopodia extend into the culture medium. PMID- 9338117 TI - The biological conditions of assessment of ethylene glycol-induced inhibition of gap junctional intercellular communication by metabolic cooperation assay. AB - Ethylene glycol (EG) has been previously shown to inhibit gap junctional intercellular communication. In this paper we examine conditions under which the effect of EG on gap junctional communication is assessed by metabolic cooperation assay. The later, after the start of metabolic cooperation assay, EG was added, the lower its inhibitory effect was. If treatment with EG began 12 h or even later after plating of cells, no significant effect on gap junctional communication was observed. Short EG treatments (2-8 h) induced a reversible inhibition of cell-to-cell communication, provided that the cells could communicate freely after the drug was removed. However, if further cell-to-cell communication was excluded, the effect of short exposures was irreversible. Using Scrape loading method we observed that after a 60 min exposure to EG the standard gap junctional intercellular communication was completely restored in a few hours. PMID- 9338119 TI - Uptake of plasmid RSV DNA by frog and mouse spermatozoa. AB - Xenopus laevis spermatozoa and mouse epididymal spermatozoa were compared in their ability to bind plasmid DNA. Spermatozoa of both species are endowed with a very similar binding capacity for plasmid pAPrC DNA carrying the complete Rous sarcoma virus (RSV) DNA. Our experiments failed to detect any substantial differences between both types of sperm cells in the kinetics of DNA binding, maximum DNA binding capacity, effect of various ions, metabolic inhibitors and substrates on DNA binding, etc. Each X. laevis and mouse sperm cell associates, on an average, with about 50 and 45 molecules, respectively, of the plasmid DNA in a DNase resistant form, if spermatozoa were exposed to the DNA at a concentration of 0.5 microgram/5 x 10(6) sperm cells. The uptake of the DNA by both types of sperm cells is strongly inhibited by heparin. The 37-kDa factor IF 1 shown by Zani et al. (1995) to specifically block DNA binding to mouse sperm cells inhibited the interaction between pAPrC DNA and frog spermatozoa with a similar intensity. PMID- 9338118 TI - Effects of recombinant rat tumor necrosis factor-alpha and interferon-gamma on the respiratory burst of rat polymorphonuclear leukocytes in whole blood. AB - Activated polymorphonuclear leukocytes (PMNL) might be an important source of host tissue damage. PMNL may be primed by various inflammatory mediators for an increased production of reactive oxygen species (ROS). The short-term priming effects of two cytokines, interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha), on rat PMNL in whole unfractionated blood were investigated in the present study. Incubation of PMNL with cytokine alone did not induce the detectable ROS production without addition of a second stimulus-phorbol myristate acetate (PMA) or opsonized zymosan particles (OZP). One-hour incubation with TNF alpha, or the mixture of TNF-alpha + IFN-gamma resulted in a dose-dependent enhancement of ROS production in response to PMA. Moreover, the incubation with both IFN-gamma and TNF-alpha caused a significantly higher increase of ROS production than with TNF-alpha alone. However, no priming effects of IFN-gamma on the PMA-induced ROS production were observed. Finally, none of the cytokines enhanced the total production of ROS upon the stimulation with OZP, although changes in the time course of ROS production were observed. Thus, different signal transduction pathways seem to be involved in PMA- and OZP-induced respiratory burst. Alternatively, partial activation and assembling of NADPH oxidase during cytokine treatment might explain the differences observed between PMA- and OZP-induced ROS production. The results of the present study suggest that IFN-gamma may enhance the priming effects of TNF-alpha. This finding may have implications for understanding the mechanisms by which both cytokines may contribute to PMNL-mediated tissue injury during various clinical conditions, as well as to host defense against invading pathogens. PMID- 9338120 TI - Infection may have shaped the evolution of the biphasic immune response to cancer. PMID- 9338121 TI - Comparative aspects of the chicken immunogenetics (review). PMID- 9338122 TI - The GT-rich sequence in the U5 region of Rous sarcoma virus long terminal repeat is required for transcription termination and 3' processing. AB - The sequences in the LTR of Rous sarcoma virus that are required for transcription termination and polyadenylation have been determined. A vector containing LTR-neo-LTR has been constructed and deletions in the U5 region of the downstream LTR have been made. The DNAs from wild-type and deletion mutant recombinant plasmids were introduced into QT6 cells and G418-resistant transformants were selected. Those transformants with neo sequences in the arrangement, LTR-neo-LTR, were analyzed for transcription termination and polyadenylation by Northern blot analysis and by S1 protection experiments. The results indicate that the polyadenylation signal, AATAAA, located in the U3 region alone, is not sufficient for 3' end processing and that the sequence between +20 and +44 in the U5 region is absolutely required for transcription termination or endonucleolytic cleavage and polyadenylation. PMID- 9338123 TI - Electron microscopic demonstration of the interaction of liposomes and cells in vitro. AB - We have investigated the interaction of liposomes with the continuous cell lines P388D1 and L-132 and mouse peritoneal macrophages. To distinguish the liposomes from other vesicular structures, we have used liposomes with incorporated protein G and gold. A heterogeneous mixture of multilamellar liposomes 30 nm up to 1000 nm in size has been employed. Samples were examined at different time intervals. We found differences in the uptake of liposomes with regard to size and rate. Cells of a P388D1 monolayer took up liposomes by endocytosis very early after addition of liposomes and the number of lysosomes in their cytoplasm increased. In L-132 cells, first a fusion occurred between liposomes and the cell cytoplasmic protrusions, in the cytoplasm of which the mitochondria had multiplied. Peritoneal macrophages phagocytosed mainly large multilamellar liposomes and the membranous system of Golgi apparatus was the most prominent structure in the cytoplasm. Phagocytosis in P388D1 and L-132 cells was noted sporadically as late as 24 h after addition of liposomes to the cells. PMID- 9338124 TI - Electron microscopic demonstration of the penetration of liposomes through skin. AB - To verify the penetration of liposomes through skin, we have used liposomes with encapsulated protein G-gold conjugate in a gel vehicle. Skin samples were examined 2, 4, 24, 48, and 72 h after liposome application. Our findings show that the penetration of liposomes through skin depends mainly on their size. Liposomes up to 600 nm in diameter penetrate through skin rather easily, whereas liposomes 1000 nm and more in diameter remain interiorized in the stratum corneum. The main penetration of liposomes proceeds along the hair sheaths as indicated by larger amounts of free liposomes in the corium of guinea pigs. PMID- 9338125 TI - Particle agglutination test "Serodia HIV-1/2" as a novel anti-HIV-1/2 screening test: comparative study on 3311 serum samples. AB - Enzyme immunoassays are most widely used screening tests for antibodies to human immunodeficiency viruses (HIV). Nevertheless, the need of simpler, noninstrumented tests is evident in many parts of the world, where laboratory facilities and trained personnel are limited, and HIV incidence is high. A recently developed variant of gelatin-particle agglutination tests, Serodia HIV 1/2 (Fujirebio Inc., Tokyo, Japan), is one of such simple and noninstrumented tests. To evaluate its utility, 3311 serum samples (281 anti-HIV-1 positive, 8 anti-HIV-2 positive and 3022 anti-HIV-1/2 negative) obtained from 2632 individuals from Slovenia, other parts of former Yugoslavia and Senegal were investigated. No false-negative results and only one false-positive result were obtained during the procedures, giving overall sensitivity and specificity of the particle agglutination test of 100% and 99.97%, respectively. We have concluded that Serodia HIV-1/2 test is highly specific and sensitive for detection of anti HIV-1/2 antibodies, suitable for small blood banks and for epidemiological surveys. PMID- 9338126 TI - Portable blood glucose monitors. AB - Portable blood glucose monitors (BGMs) play a critical role in diabetes management both in the hospital and in the home. ECRI previously evaluated BGMs in February 1992 (Health Devices 21[2]) and March 1994 (Health Devices 23[3]). For the current Evaluation, we tested 10 BGMs, rating them as appropriate for home and/or hospital use. In addition, we report updated ratings and rankings for 6 units originally evaluated in our March 1994 study. While we do not repeat results for the previously evaluated units, we do include those units in their new rating and ranking order in the Conclusions and in the Major Selection Factors table. (Because our rating and ranking scheme has been revised, we have also revised our judgments for some of the previously evaluated units.) Although all units had good or excellent accuracy and repeatability, we did find significant differences in other areas. For home use, we considered repeatability, the ease of cleaning the test area, and the ease of obtaining a reading to be the most important criteria. For hospital use, we judged units on the same criteria, as well as on the effect of hematocrit on readings and on the protection from cross-contamination between patients and between the unit and the user. This Evaluation is directed at clinicians, diabetes educators, emergency medical technicians, monitor manufacturers and suppliers, pharmacies, and individual users and their caregivers. We caution readers not to base selection or purchasing decisions on our ratings and rankings alone, but on a thorough understanding of the issues underlying our conclusions, which can be gained only by reading this Evaluation in its entirety. Readers should also consult the March 1994 issue of Health Devices for information about and our findings for units still available but not evaluated in this study. PMID- 9338127 TI - Minimum requirements for respiratory care ventilator testing. AB - Older respiratory care ventilators lack self-diagnostics, requiring that an operational verification procedure (OVP) be manually performed before each patient use. But because a formal OVP for these machines can take as long as an hour in some circumstances, there may not be time to perform one between patients. In this short Guidance Article, we provide a list of the minimum tests that should be performed on these older ventilators when a full OVP is not possible. PMID- 9338128 TI - Spotlight on ECRI's PriceGuide service. PMID- 9338129 TI - Detached tip of endoscope cleaning brush released into patient during bronchoscopy. PMID- 9338130 TI - Abbott PCA Plus II patient-controlled analgesic pumps prone to misprogramming resulting in narcotic overinfusions. PMID- 9338131 TI - Unintended inflation and deflation of Zimmer model A.T.S. 1500 pneumatic tourniquet cuffs. PMID- 9338132 TI - Improper fuse in ICS Medical NCI-480 water caloric stimulator. PMID- 9338133 TI - Thoracic impedance measurements can interfere with impedance-based rate responsive pacemakers. PMID- 9338134 TI - Phytic acid stimulates the growth of a human rhabdomyosarcoma. PMID- 9338135 TI - Osteopontin overexpression in vascular smooth muscle cells transfected with the c Ha-rasEJ oncogene. PMID- 9338136 TI - Extracellular matrix promotes differentiation of retinal pigment epithelium. PMID- 9338137 TI - Amphotericin B augments interleukin-6 production by human gingival fibroblasts in vitro. PMID- 9338139 TI - Phenotypic analysis of human fetal renal cells transformed by the SV40 large T antigen. PMID- 9338138 TI - Ambient oxygen affects uptake of 2-deoxyglucose and palmitate by rat cardiac myocytes. PMID- 9338140 TI - In vitro reconstitution of human respiratory epithelium. PMID- 9338141 TI - Mitogenic activity of keratinocyte growth factor amino-terminal truncation mutants: deletion of amino acid residues 1-15 through 1-27. PMID- 9338142 TI - Differences in nucleotide effects on intracellular pH, Na+/H+ antiport activity, and ATP-binding proteins in endothelial cells. AB - Bovine (BPAEC) and human (HPAEC) pulmonary artery endothelial cell monolayers were incubated with either ATP, ATP analogues, or UTP, followed by measurement of intracellular pH (pHi) and the rate of recovery from acidosis. ATP increased baseline pHi and the rate of acid recovery in BPAEC. This response was inhibited by the amiloride analogue, methyisobutylamiloride, demonstrating that activation of the Na+/H+ antiport was responsible for the increase in baseline pHi and the recovery from acidosis. This response had the features of both a P2Y and P2U purinergic receptor, based on the responses to a series of ATP analogues and UTP. In contrast, none of the nucleotides had any significant effect on pHi and Na+/H+ antiport activity in HPAEC. This difference in the response to extracellular nucleotides was not due to a difference in ATP metabolism between cell types, since the ectonucleotidase-resistant analogue. ATP gamma S, also had no effect on HPAEC. Analogues of cAMP had no effect on pHi or acid recovery in either cell type. Incubation of BPAEC and HPAEC with the photoaffinity ligand [32P] 8-AzATP indicated that both BPAEC and HPAEC possess an ATP-binding protein of 48 kDa. However, BPAEC exhibited an additional binding protein of 87 kDa. Thus, the contrasting response to extracellular ATP between bovine and human pulmonary artery endothelial cells may be related to differences in the signal transduction pathway leading to antiport activation, including different ATP-binding sites on the cell membrane. PMID- 9338143 TI - DNA transfection in the ecdysteroid-responsive GV1 cell line from the tobacco hornworm, Manduca sexta. AB - The embryonic cell line, GV1, from Manduca sexta was transiently transfected with DNA constructs of the Drosophila hsp70 promoter fused to either a beta galactosidase (pXH70ZT) or a chloramphenicol acetyl transferase (HSP-CAT-1) reporter gene using lipofectin. Optimal cell density, DNA:lipofectin ratio, and time of incubation were varied to determine the optimal conditions: 2 x 10(5) cells/ml, 1:3, and 5 h. Under these conditions, the transfection efficiency was about 40%. Heat inducibility of two hsp70 constructs was compared. The HSP-CAT-1, containing 1127 bp of upstream sequence, was more sensitive to heat shock than that of pXH70ZT, containing only 194 bp of upstream sequence. Thus, the 1127 bp hsp70 promoter appears to be a better inducible promoter in these cells. A 2 kb fragment of the proximal promoter region of the MHR3 gene containing a putative ecdysone response element was shown to be responsive to 20-hydroxyecdysone after its transfection into these cells. PMID- 9338144 TI - Expression of alpha-smooth muscle actin, TGF-beta 1 and TGF-beta type II receptor during connective tissue contraction. AB - Closure of rat mesenteric perforation is considered to occur by connective tissue contraction, a process that has been shown to be stimulated by transforming growth factor-beta 1. In the present study, we assessed the expression of alpha smooth muscle actin during closure by quantitative-reverse transcription polymerase chain reaction and in situ hybridization. The expression of transforming growth factor-beta 1 and transforming growth factor-beta type II receptor was also estimated in mesenteric membranes and free peritoneal cells after wounding. A larger expression of alpha-smooth muscle actin was seen around the wound edges compared to unwounded tissue. Both alpha-smooth muscle actin and transforming growth factor-beta type II receptor were expressed during Days 0, 3, 5, 7, and 10. The expression of alpha-smooth muscle actin on Day 5 was > 100 times higher than on Day 0. Transforming growth factor-beta 1 was expressed in both membranes and free peritoneal cells of unoperated control animals but down regulated after wounding, a finding that has not been reported previously. It reappeared on Days 7 and 10 in free peritoneal cells but not in perforated membranes. The enhanced expression of alpha-smooth muscle actin and down regulation of transforming growth factor-beta 1 expression after wounding appears to be important phenomena in tissue contraction and repair. PMID- 9338145 TI - Immortalization of mutant p53-transfected human fibroblasts by treatment with either 4-nitroquinoline 1-oxide or X-rays. AB - The study of in vitro cell transformation is valuable for understanding the multistep carcinogenesis of human cells. The difficulty in inducing neoplastic transformation of human cells by treatment with chemical or physical agents alone is due to the difficulty in immortalizing normal human cells. Thus, the immortalization step is critical for in vitro neoplastic transformation of human cells. We transfected a mutant p53 gene (mp53: codon 273Arg-His) into normal human fibroblasts and obtained two G418-resistant mp53-containing clones. These clones showed an extended life span but ultimately senesced. However, when they were treated with either 4-nitroquinoline 1-oxide or X-rays, they were immortalized. The immortalized cells showed both numerical and structural chromosome abnormalities, but they were not tumorigenic. The expression of mutant but not wild type p53 was detected in the immortalized cells by RT-PCR. Expression of p21, which is located downstream of p53, was remarkably reduced in the immortalized cells, resulting in increased cdk2 and cdc2 kinase activity. However, there was no significant difference between the normal and immortalized human cells in expression of another tumor suppressor gene, p16. These findings indicate that the p53-p21 cascade may play an important role in the immortalization of human cells. PMID- 9338147 TI - Multiplication of a granulosis virus isolated from the potato tuber moth in a new established cell line of Phthorimaea operculella. AB - A newly established cell line was obtained from the culture of embryonic cells of the potato tuber moth Phthorimaea operculella in low temperature conditions (19 degrees C) using modified Grace's medium supplemented with 10% fetal bovine serum. The population doubling time was about 80 h when cells were cultivated at 19 degrees C and 38 h at 27 degrees C. The cell line had a relatively homogeneous population consisting of various sized spherical cells. The cells were cultivated for more than 25 passages. Their polypeptidic profile was different from profiles of other P. operculella cell lines we previously described and from other lepidopteran cells. The new cell line was designated ORS-Pop-95. The complete replication of the potato tuber moth granulosis virus (PTM GV) was obtained in vitro by both viral infection and DNA transfection. PTM GV multiplied at a significant level during several passages of the cell line that was maintained at 19 degrees C. As long as the cells were maintained at 19 degrees C, virus multiplication could also be obtained at the same rate at 27 degrees C. To compare PTM GV multiplied both in vivo and in vitro, we used morphological identification, serological, DNA probe diagnosis and endonuclease digest profile analysis and confirmed the identity of the virus. PMID- 9338146 TI - The inability of cells to grow in low iron correlates with increasing activity of their iron regulatory protein (IRP). AB - We studied the factors that determine the differing growth requirements of low iron-tolerant (LIT) versus high-iron-dependent (HID) cells for extracellular nontransferrin iron. The growth of LIT cells HeLa and THP-1, when transferred from transferrin (5 micrograms/ml) medium into low-iron (5 microM ferric citrate) medium, was not significantly affected while HID cells Jiyoye and K562 showed nearly no growth. HeLa and THP-1 cells, as well as Jiyoye and K562 cells, do not produce transferrin in sufficient amounts to support their growth in low-iron medium. Surprisingly, similar rates of iron uptake in low-iron medium (0.033 and 0.032 nmol Fe/min and 10(6) cells) were found for LIT cells HeLa and HID cells K562. Furthermore, the intracellular iron level (4.64 nmol/10(6) cells) of HeLa cells grown in low-iron medium was much higher than iron levels (0.15 or 0.20 nmol/10(6) cells) of HeLa or K562 cells grown in transferrin medium. We demonstrated that the activity (ratio activated/total) of the iron regulatory protein (IRP) in HID cells Jiyoye and K562 increased more than twofold (from 0.32 to 0.79 and from 0.47 to 1.12, respectively) within 48 h after their transfer into low-iron medium. In the case of LIT cells HeLa and THP-1, IRP activity stayed at similar or slightly decreased levels (0.86-0.73 and 0.58-0.55, respectively). Addition of iron chelator deferoxamine (50 microM, i.e., about half-maximal growth-inhibitory dose) resulted in significantly increased activity of IRP also in HeLa and THP-1 cells. We hypothesize that the relatively higher bioavailability of nontransferrin iron in LIT cells, over that in HID cells, determines the differing responses observed under low-iron conditions. PMID- 9338408 TI - Invited editorial on "Femoral arterial injection of adenosine in humans elevates MSNA via central but not peripheral mechanisms". PMID- 9338148 TI - Characterization of proinflammatory cytokine production and CD14 expression by murine alveolar macrophage cell lines. AB - Alveolar macrophages, which play a central role in lung defense, produce cytokines that help orchestrate local inflammatory responses. In sepsis and other pathological conditions, bacterial lipopolysaccharide endotoxin can induce alveolar macrophages (AM) to release proinflammatory cytokines, including tumor necrosis factor-alpha, interleukin-1, and interleukin-6. Studying the mechanisms that control alveolar macrophage cytokine production may lead to better therapies for conditions involving inflammatory lung injury. We and others have noted significant differences between alveolar macrophages and peritoneal macrophages, but large numbers of human or murine alveolar macrophages are rarely available for detailed mechanistic studies. We have obtained three murine alveolar macrophage cell lines (AMJ2C8, AMJ2C11, and AMJ2C20) and have begun to characterize their cytokine responses to proinflammatory stimuli. We measured the effects of endotoxin, interferon gamma, and the combination of the two on production of tumor necrosis factor, interleukin-1 beta, and interleukin-6 in each line. We also studied the expression of the endotoxin receptor CD14 by these cells, and investigated the effect of serum on their endotoxin responsiveness. We show here that all three of the cell lines responded in a manner comparable to that of primary murine alveolar macrophages. Observed variations between these lines may reflect the documented heterogeneity seen in populations of primary alveolar macrophages. These cell lines should expand the repertoire of tools available to investigators studying regulation of murine alveolar macrophage responses. PMID- 9338409 TI - Femoral arterial injection of adenosine in humans elevates MSNA via central but not peripheral mechanisms. AB - The purpose of the present study was to examine the effects of femoral arterial injections of adenosine on muscle sympathetic nerve activity (MSNA) under three different conditions. These conditions were adenosine injection alone, adenosine injection after phenylephrine infusion, and adenosine injection distal to a thigh cuff inflated to arrest the circulation. The arterial injection of adenosine alone resulted in a fourfold (255 +/- 18 U/min) increase above baseline (73 +/- 12 U/min; P < 0.05) in MSNA with an onset latency of 15.8 +/- 0.8 s from the time of injection. The systemic infusion of phenylephrine resulted in an increase (P < 0.05) in mean arterial pressure of approximately 10 mmHg and a decrease (P < 0.05) in heart rate of 8-10 beats/min compared with baseline values before phenylephrine infusion. After adenosine injection, the onset latency for the increase in MSNA was delayed to 19.2 +/- 2.1 s and the magnitude of increase was attenuated by approximately 50% (123 +/- 20 U/min) compared with adenosine injection alone (P < 0.05). When a cuff was inflated to 220 mmHg to arrest the circulation and adenosine was injected into the leg distal to the inflated cuff, there were no significant changes in MSNA or any of the other measured variables. However, on deflation of the cuff, there was a rapid increase (P < 0.05) in MSNA, with an onset latency of 9.1 +/- 0.9 s, and the magnitude of increase (276 +/- 28 U/min) was similar to that observed for adenosine alone. These data suggest that approximately 50% of the effects of exogenously administered adenosine are a result of baroreceptor unloading due to a drop in blood pressure. Furthermore, the finding that adenosine did not directly result in an increase in MSNA while it was trapped in the leg but that it needed to be released into the circulation suggests that adenosine does not directly stimulate thin fiber muscle afferents in the leg of humans. In contrast, it would appear that adenosine exerts its effects via some other chemically sensitive pool of afferents. PMID- 9338410 TI - Tidal volume effects on surfactant treatment responses with the initiation of ventilation in preterm lambs. AB - We hypothesized that initiation of ventilation in preterm lambs with high volumes would cause lung injury and decrease the subsequent response to surfactant treatment. Preterm lambs were randomized to ventilation for 30 min after birth with 5 ml/kg (VT5), 10 ml/kg (VT10), or 20 ml/kg (VT20) tidal volumes and then ventilated with approximately 10 ml/kg tidal volumes to achieve arterial PCO2 values of approximately 50 Torr to 6 h of age. VT20 lambs had lower compliances, lower ventilatory efficiencies, higher recoveries of protein, and lower recoveries of surfactant in alveolar lavages and in surfactant that had decreased compliances when tested in preterm rabbits than VT5 or VT10 lambs. Other lambs randomized to treatment with surfactant at birth and ventilation with 6, 12, or 20 ml/kg tidal volumes for 30 min had no indicators of lung injury. An initial tidal volume of 20 ml/kg decreased the subsequent response to surfactant treatment, an effect that was prevented with surfactant treatment at birth. PMID- 9338411 TI - Corticosteroid effects on isotonic contractile properties of rat diaphragm muscle. AB - The effects of corticosteroids (CS) on diaphragm muscle (Diam) fiber morphology and contractile properties were evaluated in three groups of rats: controls (Ctl), surgical sham and weight-matched controls (Sham), and CS-treated (6 mg . kg-1 . day-1 prednisolone at 2.5 ml/h for 3 wk). In the CS-treated Diam, there was a selective atrophy of type IIx and IIb fibers, compared with a generalized atrophy of all fibers in the Sham group. Maximum isometric force was reduced by 20% in the CS group compared with both Ctl and Sham. Maximum shortening velocity in the CS Diam was slowed by approximately 20% compared with Ctl and Sham. Peak power output of the CS Diam was only 60% of Ctl and 70% of Sham. Endurance to repeated isotonic contractions improved in the CS-treated Diam compared with Ctl. We conclude that the atrophy of type IIx and IIb fibers in the Diam can only partially account for the CS-induced changes in isotonic contractile properties. Other factors such as reduced myofibrillar density or altered cross-bridge cycling kinetics are also likely to contribute to the effects of CS treatment. PMID- 9338412 TI - Kinematics and mechanics of midcostal diaphragm of dog. AB - Radiopaque markers were attached to the peritoneal surface of three neighboring muscle bundles in the midcostal diaphragm of four dogs, and the locations of the markers were tracked by biplanar video fluoroscopy during quiet spontaneous breathing and during inspiratory efforts against an occluded airway at three lung volumes from functional residual capacity to total lung capacity in both the prone and supine postures. Length and curvature of the muscle bundles were determined from the data on marker location. Muscle lengths for the inspiratory states, as a fraction of length at functional residual capacity, ranged from 0.89 +/- 0.04 at end inspiration during spontaneous breathing down to 0.68 +/- 0.07 during inspiratory efforts at total lung capacity. The muscle bundles were found to have the shape of circular arcs, with the three bundles forming a section of a right circular cylinder. With increasing lung volume and diaphragm displacement, the circular arcs rotate around the line of insertion on the chest wall, the arcs shorten, but the radius of curvature remains nearly constant. Maximal transdiaphragmatic pressure was calculated from muscle curvature and maximal tension-length data from the literature. The calculated maximal transdiaphragmatic pressure-length curve agrees well with the data of Road et al. (J. Appl. Physiol. 60: 63-67, 1986). PMID- 9338413 TI - Skeletal troponin I as a marker of exercise-induced muscle damage. AB - The utility of skeletal troponin I (sTnI) as a plasma marker of skeletal muscle damage after exercise was compared against creatine kinase (CK), myoglobin (Mb), and myosin heavy chain (MHC) fragments. These markers were serially measured in normal physical education teacher trainees after four different exercise regimens: 20 min of level or downhill (16% decline) running (intensity: 70% maximal O2 uptake), high-force eccentric contractions (70 repetitions), or high force isokinetic concentric contractions of the quadriceps group (40 repetitions). Eccentrically biased exercise (downhill running and eccentric contractions) promoted greater increases in all parameters. The highest plasma concentration were found after downhill running (median peaks: 309 U/l CK concentration (-CK-)), 466 microgram/l Mb concentration (-Mb-), 1,021 microU/l MHC concentration (-MHC-), and 27.3 microgram/l sTnI concentration ([sTnI]). Level running produced a moderate response (median peaks: 178 U/l -CK-, 98 microgram/l -Mb-, 501 microU/l -MHC-, and 6.6 microgram/l [sTnI]), whereas the concentric contraction protocol did not elicit significant changes in any of the markers assayed. sTnI increased and peaked in parallel to CK and stayed elevated (>2.2 microgram/l) for at least 1-2 days after exercise. In contrast to MHC, sTnI is an initial, specific marker of exercise-induced muscle injury, which may be partly explained by their different intracellular compartmentation with essentially no (MHC <0.1%) or a small soluble pool (sTnI: median 3.4%). PMID- 9338414 TI - Hemodynamic correlates of effective arterial elastance in mitral stenosis before and after balloon valvotomy. AB - This study had the purpose of documenting the hemodynamic correlates of effective arterial elastance (Ea; i.e., an accurate estimate of hydraulic load) in mitral stenosis (MS) patients. The main hypothesis tested was that Ea relates to the total vascular resistance (R)-to-pulse interval duration (T) ratio (R/T) in MS patients both before and after successful balloon mitral valvotomy (BMV). High fidelity aortic pressure recordings were obtained in 10 patients (40 +/- 12 yr) before and 15 min after BMV. Ea value was calculated as the ratio of the steady state end-systolic aortic pressure (ESAP) to stroke volume (thermodilution). Ea increased after BMV (from 1.55 +/- 0.63 to 1.83 +/- 0.71 mmHg/ml; P < 0.05). Throughout the procedure, there was a strong linear relationship between Ea and R/T: Ea = 1.09R/T - 0.01 mmHg/ml, r = 0.99, P = 0.0001. This ultimately depended on the powerful link between ESAP and mean aortic pressure [MAP; r = 0.99, 95% confidence interval for the difference (MAP - ESAP) from -18.5 to +4.5 mmHg]. Ea was also related to total arterial compliance (area method) and to wave reflections (augmentation index), although to a lesser extent. After BMV, enhanced and anticipated wave reflections were observed, and this was likely to be explained by decreased arterial compliance. The present study indicated that Ea depended mainly on the steady component of hydraulic load (i.e., R) and on heart period (i.e., T) in MS patients. PMID- 9338415 TI - Experimental murine acid aspiration injury is mediated by neutrophils and the alternative complement pathway. AB - Acid aspiration may result in the development of the acute respiratory distress syndrome, an event associated with significant morbidity and mortality. Although once attributed to direct distal airway injury, the pulmonary failure after acid aspiration is more complex and involves an inflammatory injury mediated by complement (C) and polymorphonuclear leukocytes. This study examines the injurious inflammatory cascades that are activated after acid aspiration. The role of neutrophils was defined by immunodepletion before aspiration, which reduced injury by 59%. The injury was not modified in either P- or E-selectin knockout mice, indicating that these adhesion molecules were not operative. C activation after aspiration was documented with immunochemistry by C3 deposition on injured alveolar pneumocytes. Animals in which C activation was inhibited with soluble C receptor type 1 (sCR1) had a 54% reduction in injury, similar to the level of protection seen in C3-knockout mice (58%). However C4-knockout mice were not protected from injury, indicating that C activation is mediated by the alternative pathway. Finally, an additive effect of neutrophils and C was demonstrated whereby neutropenic animals that were treated with sCR1 showed an 85% reduction in injury. Thus acid aspiration injury is mediated by neutrophils and the alternative C pathway. PMID- 9338416 TI - Trauma-induced changes of skeletal muscle membrane: decreased K+ and increased Na+ permeability. AB - Trauma of skeletal muscle causes membrane depolarization and reduces membrane resistance. The underlying mechanisms were studied in isolated mouse phrenic nerve diaphragms subject to sharp transections of muscle. Depolarization was most marked at the vicinity ( approximately 1 mm) of trauma, where the membrane potential dropped rapidly from about -80 mV to zero and repolarized to about -25 mV. At the end-plate region (located approximately 3 mm away from the cut end), the membrane gradually attained a plateau potential around -45 mV. The magnitude of depolarization was not reduced by inhibition of Na+, Ca2+, or Cl- channel, whereas the progress of depolarization was delayed in low-Na+ medium. Activation of the K+ channel with lemakalim induced some hyperpolarization at damaged site but produced a glybenclamide-sensitive outward current and hyperpolarization of end-plate region to the levels before trauma, as if there was no diminution of transmembrane K+ gradient in this area. Appropriate elevation of extracellular K+ to stimulate K+ conductance also hyperpolarized the end-plate region. The results suggest that depolarization at regions remote from trauma is related to decreased K+ and increased Na+ permeability. The cytoplasma compartmentalization and permeability changes may protect muscle fiber from trauma. PMID- 9338417 TI - Effect of exercise intensity on skeletal muscle malonyl-CoA and acetyl-CoA carboxylase. AB - Malonyl-CoA is synthesized by acetyl-CoA carboxylase (ACC) and is an inhibitor of fatty acid oxidation. Exercise induces a decline in skeletal muscle malonyl-CoA, which is accompanied by inactivation of ACC and increased activity of AMP activated protein kinase (AMPK). This study was designed to determine the effect of exercise intensity on the enzyme kinetics of ACC, malonyl-CoA levels, and AMPK activity in skeletal muscle. Male Sprague-Dawley rats were killed (pentobarbital sodium anesthesia) at rest or after 5 min of exercise (10, 20, 30, or 40 m/min at 5% grade). The fast-twitch red and white regions of the quadriceps muscle were excised and frozen in liquid nitrogen. A progressive decrease in red quadriceps ACC maximal velocity (from 28.6 +/- 1.5 to 14.3 +/- 0.7 nmol . g-1 . min-1, P < 0.05), an increase in activation constant for citrate, and a decrease in malonyl CoA (from 1.9 +/- 0.2 to 0.9 +/- 0.1 nmol/g, P < 0.05) were seen with the increase in exercise intensity from rest to 40 m/min. AMPK activity increased more than twofold. White quadriceps ACC activity decreased only during intense exercise. We conclude that the extent of ACC inactivation during short-term exercise is dependent on exercise intensity. PMID- 9338418 TI - Effects of haloperidol on ventilation during isocapnic hypoxia in humans. AB - Exposure to isocapnic hypoxia produces an abrupt increase in ventilation [acute hypoxic ventilatory response (AHVR)], which is followed by a subsequent decline [hypoxic ventilatory depression or decline (HVD)]. In cats, both anesthetized and awake, haloperidol has been reported to increase AHVR and almost entirely abolish HVD. To investigate whether this occurs in humans, the ventilatory responses of 15 healthy young volunteers to 20 min of isocapnic hypoxia (end-tidal PO2 = 50 Torr) were assessed at 1, 2, and 4.5 h after placebo (control) and after oral haloperidol (Seranace, 0.05 mg/kg) on different days. Three subjects were unable to complete the study because of akathisia. AHVR was significantly greater with haloperidol compared with control (P < 0.01, analysis of variance). However, no significant change in HVD was found [control HVD = 9.3 +/- 1.6 (SD) l/min, haloperidol HVD = 9.9 +/- 2.1 l/min; P = not significant, analysis of variance]. We conclude that combined central and peripheral dopamine-receptor antagonism in humans with haloperidol produces a similar pattern of change to that reported previously with the peripheral antagonist domperidone. We have been unable to show in humans a decrease in HVD by the centrally acting drug as observed in cats. PMID- 9338419 TI - Relationship between cold tolerance and generation of suppressor macrophages during acute cold stress. AB - Acute cold stress induces suppressor macrophages expressing large numbers of receptors to the crystallizable fragment (Fc) portion of immunoglobulin G (MAC-1+ FcgammaRII/IIIbright cells), resulting in the immunosuppression of splenocyte mitogenesis. The generation of MAC-1+ FcgammaRII/IIIbright cells is mediated by the action of glucocorticoids (GCs) through the GC-receptor. In the present study, the generation of MAC-1+ FcgammaRII/IIIbright cells in peritoneal exudate cells was closely related to the decrease of rectal temperature during 3-day exposure to 5 degrees C. We next investigated the effects of improved cold tolerance on the generation of MAC-1+ FcgammaRII/IIIbright cells during acute cold stress. Mice were adapted to cold by exposure to 5 degrees C for 3 wk (cold acclimated mice) and then reexposed to 5 degrees C for 3 h (acute cold stress) after living at 25 degrees C for 24 h. The rectal temperature of cold-acclimated mice was not decreased by the acute cold stress. In addition, the proportion of MAC-1+ FcgammaRII/IIIbright cells in peritoneal exudate cell population from cold acclimated mice was unaffected by the acute cold stress. The cold acclimation significantly attenuated the increases in serum corticosterone levels and the expression of the GC-receptor mRNA on peritoneal exudate cells in response to acute cold stress. These results suggest that the altered GC response to acute cold stress by the improvement of cold tolerance inhibits the generation of suppressor macrophages during acute cold stress. PMID- 9338420 TI - Patterns of shortening and thickening of the human diaphragm. AB - To study how the human diaphragm changes configuration during inspiration, we simultaneously measured diaphragm thickening using ultrasound and inspired volumes using a pneumotachograph. Diaphragm length was assessed by chest radiography. We found that thickening and shortening were greatest during a breath taken primarily with the abdomen. However, the degree of thickening was greater than expected for fiber shortening, assuming parallel muscle fibers and no shear. So, to clarify this unexpected finding, we considered geometric models of the diaphragm. How a muscle thickens as its fibers shorten is critically dependent on geometry. Thus, if a flat rectangular sheet of muscle shortens along one dimension, surface area-to-length ratio along this dimension should remain constant, and thickness would be inversely proportional to length during shortening. The simplest model of the diaphragm, however, is a cylindrical sheet of muscle in the zone of apposition capped by a dome; the ratio of surface area to radial fiber length in the dome is substantially less than the ratio of area to length of the cylindrical zone of apposition; hence, as the zone of apposition shortens while the dome radius remains constant, the ratio of total surface area to combined length (i.e., dome + zone of apposition) must decrease and thickening of the muscle correspondingly must increase more than expected for a simple rectangular strip. A similar relationship can be derived between thickening and length in a muscle sheet with a wedge-shaped insertion into a thin flat tendon. Comparison of calculations with these types of models to data from human subjects indicates that the unexpected thickening in the zone of apposition is explained by the peculiar geometry of the diaphragm. The greater thickening of the diaphragm in the zone of apposition suggests that more of the muscle mass and more sarcomeres are retained in the zone of apposition as the dome descends. Physiologically, this greater thickening may have importance by reducing wall stress in the zone of apposition and reducing the work or energy requirements per sarcomere. PMID- 9338421 TI - Rate and composition of sweat fluid losses are unaltered by hypohydration during prolonged exercise in horses. AB - Rate and ionic composition of sweat fluid losses and partitioning of evaporative heat loss into respiratory and cutaneous components were determined in six horses during three 15-km phases of exercise at approximately 40% of maximal O2 uptake. Pattern of change in sweat rate (SR) and composition was similar during each phase. SR increased rapidly for the first 20 min of exercise but remained at approximately 24-28 ml . m-2 . min-1 during the remainder of each phase. Similarly, the concentrations of Na and Cl in sweat increased until 30 min of exercise but were unchanged thereafter. Sweat osmolality and concentrations of Na and Cl were positively correlated with SR. Sweat K concentration decreased during exercise but was not correlated with SR. Fluid losses were 33.8 +/- 1.5 liters, resulting in decreases of approximately 21% in plasma volume and approximately 11% in total body water. The approximately 6% hypohydration was not associated with an alteration in SR, sweat composition, or heat storage. Respiratory and cutaneous evaporative heat loss represented approximately 23 and 70%, respectively, of the total heat dissipated, and the partitioning of heat loss was similar in each exercise phase. We conclude that SR and the relative proportions of respiratory and cutaneous evaporative heat loss are unchanged in horses during prolonged low-intensity exercise despite moderate hypohydration. PMID- 9338422 TI - Effects of bovine colostrum supplementation on serum IGF-I, IgG, hormone, and saliva IgA during training. AB - The purpose of this study was to examine the effects of bovine colostrum supplementation (Bioenervi) on serum insulin-like growth factor I (IGF-I), immunoglobulin G, hormone, and amino acid and saliva immunoglobulin A concentrations during a strength and speed training period. Nine male sprinters and jumpers underwent three randomized experimental training treatments of 8 days separated by 13 days. The only difference in the treatments was the drink of 125 ml consumed per day. Posttraining increases were noticed for serum IGF-I in the 25-ml Bioenervi treatment (125 ml contained 25 ml Bioenervi) and especially in the 125-ml Bioenervi treatment (125 ml contained 125 ml Bioenervi) compared with the placebo (normal milk whey) treatment (P < 0.05). The change in IGF-I concentration during the 8-day periods correlated positively with the change in insulin concentration during the same periods with 25-ml Bioenervi treatment (r = 0.68; P = 0.045) and with 125-ml Bioenervi treatment (r = 0.69; P = 0.038). Serum immunoglobulin G, hormone, and amino acid and saliva immunoglobulin A responses were similar during the three treatments. It appears that a bovine colostrum supplement (Bioenervi) may increase serum IGF-I concentration in athletes during strength and speed training. PMID- 9338423 TI - Restoration of fluid balance after exercise-induced dehydration: effects of alcohol consumption. AB - The effect of alcohol consumption on the restoration of fluid and electrolyte balance after exercise-induced dehydration [2.01 +/- 0.10% (SD) of body mass] was investigated. Drinks containing 0, 1, 2, and 4% alcohol were consumed over 60 min beginning 30 min after the end of exercise; a different beverage was consumed in each of four trials. The volume consumed (2,212 +/- 153 ml) was equivalent to 150% of body mass loss. Peak urine flow rate occurred later (P = 0.024) with the 4% beverage. The total volume of urine produced over the 6 h after rehydration, although not different between trials (P = 0.307), tended to increase as the quantity of alcohol ingested increased. The increase in blood (P = 0.013) and plasma (P = 0.050) volume with rehydration was slower when the 4% beverage was consumed and did not increase to values significantly greater than the dehydrated level (P = 0.013 and P = 0.050 for blood volume and plasma volume, respectively); generally, the increase was an inverse function of the quantity of alcohol consumed. These results suggest that alcohol has a negligible diuretic effect when consumed in dilute solution after a moderate level of hypohydration induced by exercise in the heat. There appears to be no difference in recovery from dehydration whether the rehydration beverage is alcohol free or contains up to 2% alcohol, but drinks containing 4% alcohol tend to delay the recovery process. PMID- 9338424 TI - A bout of resistance exercise increases urinary calcium independently of osteoclastic activation in men. AB - Metabolic acidosis increases urinary calcium excretion in humans as a result of administration of ammonium chloride, an increase in dietary protein intake, and fasting-induced ketoacidosis. An intense bout of exercise, exceeding aerobic capacity, also causes significant decrease in blood pH as a result of increase in blood lactate concentration. In this study we investigated changes in renal calcium handling, plasma parathyroid hormone concentration, and osteoclastic bone resorption after a single bout of resistance exercise. Ten male subjects completed a bout of resistance exercise with an intensity of 60% of one repetition maximum for the first set and 80% of one repetition maximum for the second and third sets. After exercise, blood and urine pH shifted toward acidity and urinary calcium excretion increased. Hypercalciuria was observed in the presence of an increased fractional calcium excretion and an unchanged filtered load of calcium. Therefore, the observed increase in urinary calcium excretion was due primarily to decrease in renal tubular reabsorption of calcium. Likely causes of the increase in renal excretion of calcium are metabolic acidosis itself and decreased parathyroid hormone. When urinary calcium excretion increased, urinary deoxypyridinoline, a marker of osteoclastic bone resorption, decreased. These results suggest that 1) strenuous resistance exercise increased urinary calcium excretion by decreasing renal tubular calcium reabsorption, 2) urinary calcium excretion increased independently of osteoclast activation, and 3) the mechanism resulting in postexercise hypercalciuria might involve non-cell mediated physicochemical bone dissolution. PMID- 9338426 TI - Classic conditioning of the ventilatory responses in rats. AB - Recent authors have stressed the role of conditioning in the control of breathing, but experimental evidence of this role is still sparse and contradictory. To establish that classic conditioning of the ventilatory responses can occur in rats, we performed a controlled experiment in which a 1 min tone [conditioned stimulus (CS)] was paired with a hypercapnic stimulus [8.5% CO2, unconditioned stimulus (US)]. The experimental group (n = 9) received five paired CS-US presentations, followed by one CS alone to test conditioning. This sequence was repeated six times. The control group (n = 7) received the same number of CS and US, but each US was delivered 3 min after the CS. We observed that after the CS alone, breath duration was significantly longer in the experimental than in the control group and mean ventilation was significantly lower, thus showing inhibitory conditioning. This conditioning may have resulted from the association between the CS and the inhibitory and aversive effects of CO2. The present results confirmed the high sensitivity of the respiratory controller to conditioning processes. PMID- 9338425 TI - Effects of nitric oxide on blood flow distribution and O2 extraction capabilities during endotoxic shock. AB - The effects of the nitric oxide (NO) synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) and the NO donor 3-morpholinosydnonimine (SIN-1) were tested in 18 endotoxic dogs. L-NMMA infusion (10 mg . kg-1 . h-1) increased arterial and pulmonary artery pressures and systemic and pulmonary vascular resistances but decreased cardiac index, left ventricular stroke work index, and blood flow to the hepatic, portal, mesenteric, and renal beds. SIN-1 infusion (2 microg . kg-1 . min-1) increased cardiac index; left ventricular stroke work index; and hepatic, portal, and mesenteric blood flow. It did not significantly influence arterial and pulmonary artery pressures but decreased renal blood flow. The critical O2 delivery was similar in the L-NMMA group and in the control group (13.3 +/- 1.6 vs. 12.8 +/- 3.3 ml . kg-1 . min-1) but lower in the SIN-1 group (9.1 +/- 1.8 ml . kg-1 . min-1, both P < 0.05). The critical O2 extraction ratio was also higher in the SIN-1 group than in the other groups (58.7 +/- 10.6 vs. 42.2 +/- 7.6% in controls, P < 0.05; 43.0 +/- 15.5% in L-NMMA group, P = not significant). We conclude that NO is not implicated in the alterations in O2 extraction capabilities observed early after endotoxin administration. PMID- 9338428 TI - Relationship between heterogeneous changes in airway morphometry and lung resistance and elastance. AB - We present a dog lung model to predict the relation between inhomogeneous changes in airway morphometry and lung resistance (RL) and elastance (EL) for frequencies surrounding typical breathing rates. The RL and EL were sensitive in distinct ways to two forms of peripheral constriction. First, when there is a large and homogeneous constriction, the RL increases uniformly over the frequency range. The EL is rather unaffected below 1 Hz but then increases with frequencies up to 5 Hz. This increase is caused by central airway wall shunting. Second, the RL and EL are extremely sensitive to mild inhomogeneous constriction in which a few highly constricted or nearly closed airways occur randomly throughout the periphery. This results in extreme increases in the levels and frequency dependence of RL and EL but predominantly at typical breathing rates (<1 Hz). Conversely, the RL and EL are insensitive to highly inhomogeneous airway constriction that does not produce any nearly closed airways. Similarly, alterations in the RL and EL due to central airway wall shunting are not likely until the preponderance of the periphery constricts substantially. The RL and EL spectra are far more sensitive to these two forms of peripheral constriction than to constriction conditions known to occur in the central airways. On the basis of these simulations, we derived a set of qualitative criteria to infer airway constriction conditions from RL and EL spectra. PMID- 9338429 TI - Effect of tidal volume and frequency on airway responsiveness in mechanically ventilated rabbits. AB - We evaluated the effects of the rate and volume of tidal ventilation on airway resistance (Raw) during intravenous methacholine (MCh) challenge in mechanically ventilated rabbits. Five rabbits were challenged at tidal volumes of 5, 10, and 20 ml/kg at a frequency of 15 breaths/min and also under static conditions (0 ml/kg tidal volume). Four rabbits were subjected to MCh challenge at frequencies of 6 and 30 breaths/min with a tidal volume of 10 ml/kg and also under static conditions. In both groups, the increase in Raw with MCh challenge was significantly greater under static conditions than during tidal ventilation at any frequency or volume. Increases in the volume or frequency of tidal ventilation resulted in significant decreases in Raw in response to MCh. We conclude that tidal breathing suppresses airway responsiveness in rabbits in vivo. The suppression of narrowing in response to MCh increases as the magnitude of the volume or the frequency of the tidal oscillations is increased. Our findings suggest that the effect of lung volume changes on airway responsiveness in vivo is primarily related to the stretch of airway smooth muscle. PMID- 9338427 TI - Hemodynamics, cerebral circulation, and oxygen saturation in Cheyne-Stokes respiration. AB - Because cardiovascular disorders and stroke may induce Cheyne-Stokes respiration, our purpose was to study the interaction among cerebral activity, cerebral circulation, blood pressure, and blood gases during Cheyne-Stokes respiration. Ten patients with heart failure or a previous stroke were investigated during Cheyne-Stokes respiration with recordings of daytime polysomnography, cerebral blood flow velocity, intra-arterial blood pressure, and intra-arterial oxygen saturation with and without oxygen administration. There were simultaneous changes in wakefulness, cerebral blood flow velocity, and respiration with accompanying changes in blood pressure and heart rate approximately 10 s later. Cerebral blood flow velocity, blood pressure, and heart rate had a minimum occurrence in apnea and a maximum occurrence during hyperpnea. The apnea-induced oxygen desaturations were diminished during oxygen administration, but the hemodynamic alterations persisted. Oxygen desaturations were more severe and occurred earlier according to intra-arterial measurements than with finger oximetry. It is not possible to explain Cheyne-Stokes respiration by alterations in blood gases and circulatory time alone. Cheyne-Stokes respiration may be characterized as a state of phase-linked cyclic changes in cerebral, respiratory, and cardiovascular functions probably generated by variations in central nervous activity. PMID- 9338431 TI - Lysophosphatidic acid enhances contractility of isolated airway smooth muscle. AB - The effects of the simple phospholipid mediator lysophosphatidic acid (LPA) on the contractile responsiveness of isolated tracheal rings from rabbits and cats were assessed. In both species, LPA increased the contractile response to the muscarinic agonist methacholine, but LPA did not induce contraction on its own. Conversely, LPA decreased the relaxation response to the beta-adrenergic-agonist isoproterenol in both species. Concentrations of LPA as low as 10(-8) M were effective, and the effects of LPA were rapidly reversed on washing. Phosphatidic acid was much less effective, requiring higher concentrations and producing only a minimal effect. Contractions induced by serotonin and by substance P also were enhanced by LPA, but KCl-induced contractions were unaffected. LPA inhibited the isoproterenol-induced relaxation of KCl-precontracted rings, similar to its effects on methacholine-precontracted rings, and relaxation induced by the direct adenylyl cyclase activator forskolin was inhibited in a manner similar to that induced by isoproterenol. Epithelium removal did not alter the contraction enhancing effect of LPA. The ability of LPA to both enhance contraction and inhibit relaxation of airway smooth muscle suggests that LPA could contribute to airway hypercontractility in asthma, airway inflammation, or other types of lung injury. PMID- 9338430 TI - Nonspecific endothelin-receptor antagonist blunts monocrotaline-induced pulmonary hypertension in rats. AB - Endothelin-1 (ET-1), a potent vasoactive and mitogenic peptide, has been implicated in the pathogenesis of several forms of pulmonary hypertension. We hypothesized that nonspecific blockade of ET receptors would blunt the development of monocrotaline (MCT)-induced pulmonary hypertension in rats. A single dose of the nonspecific ET blocker bosentan (100 mg/kg) given to intact rats by gavage completely blocked the pulmonary vasoconstrictor actions of Big ET 1 and partially blunted hypoxic pulmonary vasoconstriction. After 3 wk, MCT injected (105 mg/kg sc) rats gavaged once daily with bosentan (200 mg/kg) had lower right ventricular (RV) systolic pressure (RVSP), RV-to-body weight (RV/BW) and RV-to-left ventricular (LV) plus septal (S) weight [RV/(LV+S)] ratios and less percent medial thickness of small pulmonary arteries than control MCT injected rats. Lower dose bosentan (100 mg/kg) had no effect on these parameters after MCT or saline injection. Bosentan raised plasma ET-1 levels but had no effect on lung ET-1 levels. Bosentan (200 mg/kg) also had no effect on wet-to-dry lung weight ratios 6 days after MCT injection. When given during the last 10 days, but not the first 11 days of a 3-wk period after MCT injection, bosentan reduced RV/(LV+S) compared with MCT-injected controls. We conclude that ET-1 contributes to the pathogenesis of MCT-induced pulmonary hypertension and acts mainly during the later inflammatory rather than the acute injury phase after injection. PMID- 9338432 TI - Simulation of motor unit recruitment and microvascular unit perfusion: spatial considerations. AB - Muscle fiber activity is the principal stimulus for increasing capillary perfusion during exercise. The control elements of perfusion, i.e., microvascular units (MVUs), supply clusters of muscle fibers, whereas the control elements of contraction, i.e., motor units, are composed of fibers widely scattered throughout muscle. The purpose of this study was to examine how the discordant spatial domains of MVUs and motor units could influence the proportion of open capillaries (designated as perfusion) throughout a muscle cross section. A computer model simulated the locations of perfused MVUs in response to the activation of up to 100 motor units in a muscle with 40,000 fibers and a cross sectional area of 100 mm2. The simulation increased contraction intensity by progressive recruitment of motor units. For each step of motor unit recruitment, the percentage of active fibers and the number of perfused MVUs were determined for several conditions: 1) motor unit fibers widely dispersed and motor unit territories randomly located (which approximates healthy human muscle), 2) regionalized motor unit territories, 3) reversed recruitment order of motor units, 4) densely clustered motor unit fibers, and 5) increased size but decreased number of motor units. The simulations indicated that the widespread dispersion of motor unit fibers facilitates complete capillary (MVU) perfusion of muscle at low levels of activity. The efficacy by which muscle fiber activity induced perfusion was reduced 7- to 14-fold under conditions that decreased the dispersion of active fibers, increased the size of motor units, or reversed the sequence of motor unit recruitment. Such conditions are similar to those that arise in neuromuscular disorders, with aging, or during electrical stimulation of muscle, respectively. PMID- 9338433 TI - VO2 kinetics in the horse during moderate and heavy exercise. AB - The horse is a superb athlete, achieving a maximal O2 uptake (approximately 160 ml . min-1 . kg-1) approaching twice that of the fittest humans. Although equine O2 uptake (VO2) kinetics are reportedly fast, they have not been precisely characterized, nor has their exercise intensity dependence been elucidated. To address these issues, adult male horses underwent incremental treadmill testing to determine their lactate threshold (Tlac) and peak VO2 (VO2 peak), and kinetic features of their VO2 response to "square-wave" work forcings were resolved using exercise transitions from 3 m/s to a below-Tlac speed of 7 m/s or an above-Tlac speed of 12.3 +/- 0.7 m/s (i.e., between Tlac and VO2 peak) sustained for 6 min. VO2 and CO2 output were measured using an open-flow system: pulmonary artery temperature was monitored, and mixed venous blood was sampled for plasma lactate. VO2 kinetics at work levels below Tlac were well fit by a two-phase exponential model, with a phase 2 time constant (tau1 = 10.0 +/- 0.9 s) that followed a time delay (TD1 = 18.9 +/- 1.9 s). TD1 was similar to that found in humans performing leg cycling exercise, but the time constant was substantially faster. For speeds above Tlac, TD1 was unchanged (20.3 +/- 1.2 s); however, the phase 2 time constant was significantly slower (tau1 = 20.7 +/- 3.4 s, P < 0.05) than for exercise below Tlac. Furthermore, in four of five horses, a secondary, delayed increase in VO2 became evident 135.7 +/- 28.5 s after the exercise transition. This "slow component" accounted for approximately 12% (5.8 +/- 2.7 l/min) of the net increase in exercise VO2. We conclude that, at exercise intensities below and above Tlac, qualitative features of VO2 kinetics in the horse are similar to those in humans. However, at speeds below Tlac the fast component of the response is more rapid than that reported for humans, likely reflecting different energetics of O2 utilization within equine muscle fibers. PMID- 9338434 TI - Rib cage mechanics during quiet breathing and exercise in humans. AB - During exercise, large pleural, abdominal, and transdiaphragmatic pressure swings might produce substantial rib cage (RC) distortions. We used a three-compartment chest wall model (J. Appl. Physiol. 72: 1338-1347, 1992) to measure distortions of lung- and diaphragm-apposed RC compartments (RCp and RCa) along with pleural and abdominal pressures in five normal men. RCp and RCa volumes were calculated from three-dimensional locations of 86 markers on the chest wall, and the undistorted (relaxation) RC configuration was measured. Compliances of RCp and RCa measured during phrenic stimulation against a closed airway were 20 and 0%, respectively, of their values during relaxation. There was marked RC distortion. Thus nonuniform distribution of pressures distorts the RC and markedly stiffens it. However, during steady-state ergometer exercise at 0, 30, 50, and 70% of maximum workload, RC distortions were small because of a coordinated action of respiratory muscles, so that net pressures acting on RCp and RCa were nearly the same throughout the respiratory cycle. This maximizes RC compliance and minimizes the work of RC displacement. During quiet breathing, plots of RCa volume vs. abdominal pressure were to the right of the relaxation curve, indicating an expiratory action on RCa. We attribute this to passive stretching of abdominal muscles, which more than counterbalances the insertional component of transdiaphragmatic pressure. PMID- 9338435 TI - Human respiratory muscle actions and control during exercise. AB - We measured pressures and power of diaphragm, rib cage, and abdominal muscles during quiet breathing (QB) and exercise at 0, 30, 50, and 70% maximum workload (Wmax) in five men. By three-dimensional tracking of 86 chest wall markers, we calculated the volumes of lung- and diaphragm-apposed rib cage compartments (Vrc,p and Vrc,a, respectively) and the abdomen (Vab). End-inspiratory lung volume increased with percentage of Wmax as a result of an increase in Vrc,p and Vrc,a. End-expiratory lung volume decreased as a result of a decrease in Vab. DeltaVrc,a/DeltaVab was constant and independent of Wmax. Thus we used DeltaVab/time as an index of diaphragm velocity of shortening. From QB to 70% Wmax, diaphragmatic pressure (Pdi) increased approximately 2-fold, diaphragm velocity of shortening 6.5-fold, and diaphragm workload 13-fold. Abdominal muscle pressure was approximately 0 during QB but was equal to and 180 degrees out of phase with rib cage muscle pressure at all percent Wmax. Rib cage muscle pressure and abdominal muscle pressure were greater than Pdi, but the ratios of these pressures were constant. There was a gradual inspiratory relaxation of abdominal muscles, causing abdominal pressure to fall, which minimized Pdi and decreased the expiratory action of the abdominal muscles on Vrc,a gradually, minimizing rib cage distortions. We conclude that from QB to 0% Wmax there is a switch in respiratory muscle control, with immediate recruitment of rib cage and abdominal muscles. Thereafter, a simple mechanism that increases drive equally to all three muscle groups, with drive to abdominal and rib cage muscles 180 degrees out of phase, allows the diaphragm to contract quasi-isotonically and act as a flow generator, while rib cage and abdominal muscles develop the pressures to displace the rib cage and abdomen, respectively. This acts to equalize the pressures acting on both rib cage compartments, minimizing rib cage distortion. PMID- 9338436 TI - Myogenic regulatory factors during regeneration of skeletal muscle in young, adult, and old rats. AB - Myogenic factor mRNA expression was examined during muscle regeneration after bupivacaine injection in Fischer 344/Brown Norway F1 rats aged 3, 18, and 31 mo of age (young, adult, and old, respectively). Mass of the tibialis anterior muscle in the young rats had recovered to control values by 21 days postbupivacaine injection but in adult and old rats remained 40% less than that of contralateral controls at 21 and 28 days of recovery. During muscle regeneration, myogenin mRNA was significantly increased in muscles of young, adult, and old rats 5 days after bupivacaine injection. Subsequently, myogenin mRNA levels in young rat muscle decreased to postinjection control values by day 21 but did not return to control values in 28-day regenerating muscles of adult and old rats. The expression of MyoD mRNA was also increased in muscles at day 5 of regeneration in young, adult, and old rats, decreased to control levels by day 14 in young and adult rats, and remained elevated in the old rats for 28 days. In summary, either a diminished ability to downregulate myogenin and MyoD mRNAs in regenerating muscle occurs in old rat muscles, or the continuing myogenic effort includes elevated expression of these mRNAs. PMID- 9338437 TI - Passive sensitization of human airways induces myogenic contractile responses in vitro. AB - We assessed effects of passive sensitization on human bronchial smooth muscle (BSM) response to mechanical stretching in vitro. Bronchial rings were sham (control) or passively sensitized overnight by using sera from donors demonstrating sensitivity to Dermatophagoides farinae and having immunoglobulin E (IgE) concentrations of 2,600 +/- 200 U/ml. Tissues were fixed isometrically to force transducers to measure responses to electrical field stimulation (EFS) and quick stretch (QS). The myogenic response to QS was normalized to the maximal response to EFS (%EFS). The myogenic response of sensitized BSM was 47.9 +/- 10.9 %EFS to a QS of approximately 6.5% optimal length (Lo); sham-sensitized tissues had a myogenic response of 13.5 +/- 6.4 %EFS (P = 0.012 vs. passively sensitized). A QS of approximately 13% Lo in sensitized BSM caused a response of 82.8 +/- 20.9 %EFS; sham-sensitized tissues developed a response of 38.2 +/- 17.3 %EFS (P = 0.004). BSM incubated with serum from nonallergic donors did not demonstrate increased QS response (4.6 +/- 1.4 %EFS, P = not significant vs. tissue exposed to atopic sera). However, tissues incubated in sera from nonatopic donors supplemented with hapten-specific chimeric IgE (JW8) demonstrated augmented myogenic response to QS of approximately 6.5% Lo (21.9 +/- 6.2 %EFS, P = 0. 027 vs. nonatopic sera alone). We demonstrate that passive sensitization of human BSM preparations causes induction and augmentation of myogenic contractions to QS; this hyperresponsiveness corresponds to the IgE concentration in sensitizing sera. PMID- 9338438 TI - Cardiovascular changes during deep breath-hold dives in a pressure chamber. AB - Electrocardiogram, cardiac output, and blood lactate accumulation were recorded in three elite breath-hold divers diving to 40-55 m in a pressure chamber in thermoneutral (35 degrees C) or cool (25 degrees C) water. In two of the divers, invasive recordings of arterial blood pressure were also obtained during dives to 50 m in cool water. Bradycardia during the dives was more pronounced and developed more rapidly in the cool water, with heart rates dropping to 20-30 beats/min. Arrhythmias occurred, particularly during the dives in cool water, when they were often more frequent than sinus beats. Because of bradycardia, cardiac output decreased during the dives, especially in cool water (to <3 l/min in 2 of the divers). Arterial blood pressure increased dramatically, reaching values as high as 280/200 and 290/150 mmHg in the two divers, respectively. This hypertension was secondary to peripheral vasoconstriction, which also led to anaerobic metabolism, reflected in increased blood lactate concentration. The diving response of these divers resembles the one described for diving animals, although the presence of arrhythmias and large increases in blood pressure indicate a less perfect adaptation in humans. PMID- 9338439 TI - Changes in skeletal muscle biochemistry and histology relative to fiber type in rats with heart failure. AB - One of the primary consequences of left ventricular dysfunction (LVD) after myocardial infarction is a decrement in exercise capacity. Several factors have been hypothesized to account for this decrement, including alterations in skeletal muscle metabolism and aerobic capacity. The purpose of this study was to determine whether LVD-induced alterations in skeletal muscle enzyme activities, fiber composition, and fiber size are 1) generalized in muscles or specific to muscles composed primarily of a given fiber type and 2) related to the severity of the LVD. Female Wistar rats were divided into three groups: sham-operated controls (n = 13) and rats with moderate (n = 10) and severe (n = 7) LVD. LVD was surgically induced by ligating the left main coronary artery and resulted in elevations (P < 0.05) in left ventricular end-diastolic pressure (sham, 5 +/- 1 mmHg; moderate LVD, 11 +/- 1 mmHg; severe LVD, 25 +/- 1 mmHg). Moderate LVD decreased the activities of phosphofructokinase (PFK) and citrate synthase in one muscle composed of type IIB fibers but did not modify fiber composition or size of any muscle studied. However, severe LVD diminished the activity of enzymes involved in terminal and beta-oxidation in muscles composed primarily of type I fibers, type IIA fibers, and type IIB fibers. In addition, severe LVD induced a reduction in the activity of PFK in type IIB muscle, a 10% reduction in the percentage of type IID/X fibers, and a corresponding increase in the portion of type IIB fibers. Atrophy of type I fibers, type IIA fibers, and/or type IIB fibers occurred in soleus and plantaris muscles of rats with severe LVD. These data indicate that rats with severe LVD after myocardial infarction exhibit 1) decrements in mitochondrial enzyme activities independent of muscle fiber composition, 2) a reduction in PFK activity in type IIB muscle, 3) transformation of type IID/X to type IIB fibers, and 4) atrophy of type I, IIA, and IIB fibers. PMID- 9338440 TI - Effects of endurance training on the cardiovascular system and water compartments in elderly subjects. AB - The effects of endurance training on the water compartments and the cardiovascular system were determined in 10 elderly subjects [age 62 +/- 2 yr, pretraining maximal oxygen consumption (VO2 max)/kg = 25 +/- 2 ml . min-1 . kg-1 body wt]. They trained on a cycloergometer 3 times/wk for 16 wk (50-80% VO2 max, then 80-85% VO2 max). They were checked at 8 wk, 16 wk, and 4 mo after detraining. Training improved VO2 max (+16%) and induced plasma volume expansion (+11%). No change in total body water, extracellular fluid, interstitial and intracellular fluid volumes, fat-free mass, and body weight was detected in this small sample with training. Body fat mass decreased (-2.1 +/- 2.2 kg). Echocardiography at rest showed increased fractional shortening and ejection fraction and decreased left ventricular end-systolic dimension (P < 0.05). Blood volume expansion correlates with cardiac contractility and has an impact on cardiac function. These improvements are precarious, however, and are completely lost after 4 mo of detraining, when elderly subjects lose the constraints and the social stimulation of the imposed protocol. PMID- 9338441 TI - Role of neutrophils in lung vascular injury and edema after premature birth in lambs. AB - To investigate the role of neutrophils in the pathogenesis of respiratory distress after premature birth, we assessed the relationship between circulating neutrophil concentration and neutrophil accumulation in the lung, lung lymph and pleural liquid flow, and extravascular lung water in 10 chronically catheterized preterm lambs (127 +/- 1 days gestation) that were mechanically ventilated for 8 h after birth. Circulating neutrophil concentration transiently decreased within 2 h after birth and then returned to prenatal values by 6-8 h. The decrease in circulating neutrophil concentration was related directly to the accumulation of neutrophils in the air spaces, drainage of liquid and protein from the lung 6-8 h after delivery, and postmortem extravascular lung water. In additional studies, we intravenously administered mechlorethamine to 5 fetal lambs to reduce circulating neutrophils before delivery (neutrophil concentration before birth: 9 +/- 11 cells/microl). Compared with control lambs, neutrophil-depleted lambs had significantly less drainage of liquid (7.8 +/- 5.9 vs. 2.6 +/- 1.9 ml/h, respectively) and protein (116 +/- 74 vs. 42 +/- 27 mg/h, respectively) from the lung 6-8 h after birth and significantly less extravascular lung water at postmortem (6.5 +/- 0. 8 vs. 4.8 0.6 g/g dry lung, respectively). Thus neutrophils contribute to the pathogenesis of respiratory distress after premature birth by increasing lung vascular protein permeability and promoting lung edema. PMID- 9338442 TI - Acceleration of VO2 kinetics in heavy submaximal exercise by hyperoxia and prior high-intensity exercise. AB - We examined the hypothesis that O2 uptake (VO2) would change more rapidly at the onset of step work rate transitions in exercise with hyperoxic gas breathing and after prior high-intensity exercise. The kinetics of VO2 were determined from the mean response time (MRT; time to 63% of total change in VO2) and calculations of O2 deficit and slow component during normoxic and hyperoxic gas breathing in one group of seven subjects during exercise below and above ventilatory threshold (VT) and in another group of seven subjects during exercise above VT with and without prior high-intensity exercise. In exercise transitions below VT, hyperoxic gas breathing did not affect the kinetic response of VO2 at the onset or end of exercise. At work rates above VT, hyperoxic gas breathing accelerated both the on- and off-transient MRT, reduced the O2 deficit, and decreased the VO2 slow component from minute 3 to minute 6 of exercise, compared with normoxia. Prior exercise above VT accelerated the on-transient MRT and reduced the VO2 slow component from minute 3 to minute 6 of exercise in a second bout of exercise with both normoxic and hyperoxic gas breathing. However, the summated O2 deficit in the second normoxic and hyperoxic steps was not different from that of the first steps in the same gas condition. Faster on-transient responses in exercise above, but not below, VT with hyperoxia and, to a lesser degree, after prior high intensity exercise above VT support the theory of an O2 transport limitation at the onset of exercise for workloads >VT. PMID- 9338443 TI - Skeletal muscle force and actomyosin ATPase activity reduced by nitric oxide donor. AB - Nitric oxide (NO) may exert direct effects on actin-myosin cross-bridge cycling by modulating critical thiols on the myosin head. In the present study, the effects of the NO donor sodium nitroprusside (SNP; 100 microM to 10 mM) on mechanical properties and actomyosin adenosinetriphosphatase (ATPase) activity of single permeabilized muscle fibers from the rabbit psoas muscle were determined. The effects of N-ethylmaleimide (NEM; 5-250 microM), a thiol-specific alkylating reagent, on mechanical properties of single fibers were also evaluated. Both NEM (>/=25 microM) and SNP (>/=1 mM) significantly inhibited isometric force and actomyosin ATPase activity. The unloaded shortening velocity of SNP-treated single fibers was decreased, but to a lesser extent, suggesting that SNP effects on isometric force and actomyosin ATPase were largely due to decreased cross bridge recruitment. The calcium sensitivity of SNP-treated single fibers was also decreased. The effects of SNP, but not NEM, on force and actomyosin ATPase activity were reversed by treatment with 10 mM DL-dithiothreitol, a thiol reducing agent. We conclude that the NO donor SNP inhibits contractile function caused by reversible oxidation of contractile protein thiols. PMID- 9338444 TI - Effect of increased muscle temperature on oxygen uptake kinetics during exercise. AB - To test whether increased muscle temperature (Tm) would improve O2 uptake (VO2) kinetics, seven men performed transitions from rest to a moderate work rate [below the estimated lactate threshold (LTest)] and a heavy work rate (VO2 = 50% of the difference between LTest and peak VO2) under conditions of normal Tm (N) and increased Tm (H), produced by wearing hot water-perfused pants before exercise. Quadriceps Tm was significantly higher in H, but rectal temperature was similar for the two conditions. There were no significant differences in the amplitudes of the fast component of VO2 or in the time constants of the on and off transients for moderate and heavy exercise between the two conditions. The increment in VO2 between the 3rd and 6th min of heavy exercise was slightly but significantly smaller for H than for N. These data suggest that elevated Tm before exercise onset, which would have been expected to increase O2 delivery and off-loading to the muscle, had no appreciable effect on the fast exponential component of VO2 kinetics (invariant time constant). These data further suggest that elevated Tm does not contribute to the slow component of VO2 during heavy exercise. PMID- 9338445 TI - Gut and liver fat metabolism in depancreatized dogs: effects of exercise and acute insulin infusion. AB - Excessive circulating fat levels are a defining feature of poor metabolic control in diabetes. Splanchnic adipose tissue is a source of free fatty acids (FFA), and the liver is a key site of FFA utilization and the sole source of ketones. Despite the role of splanchnic tissues in fat metabolism, little is known about how these tissues respond to diabetes under divergent metabolic conditions. Therefore, splanchnic fat metabolism was studied in poorly controlled diabetes under two conditions. First, it was studied during exercise, a stimulus that enhances FFA flux. Second, it was studied while insulin was being acutely infused to achieve levels normally present during exercise, a treatment that may be expected to inhibit lipolysis. For this purpose, liver and gut arteriovenous differences were used during rest and 2.5 h of treadmill exercise in insulin deficient (n = 6) and acutely insulin-infused (n = 4) depancreatized (PX) dogs. The data show that 1) exercise, in insulin-deficient PX dogs, leads to an increase in net FFA release from mesenteric fat that is equal in magnitude to the response in nondiabetic dogs; 2) net hepatic fractional FFA extraction is increased twofold during exercise in both insulin-deficient PX dogs and nondiabetic control dogs; 3) during exercise, approximately 40 and 75% of the FFA consumed by the liver is effectively transferred from fat stores mobilized from splanchnic adipose tissue in insulin-deficient PX and nondiabetic dogs, respectively; 4) hepatic ketogenic efficiency is elevated during rest three- to fourfold in insulin-deficient PX dogs compared with nondiabetic control dogs and remains elevated during exercise; and 5) surprisingly, acute insulin replacement is ineffective in normalizing net gut, hepatic, or splanchnic FFA or ketone body balances in PX dogs. PMID- 9338446 TI - Interaction between ion transporters and the mucociliary transport system in dog and baboon. AB - To gain insight into the role of epithelial ion channels, pumps, and cotransporters in regulating airway water and mucociliary transport, we administered inhibitors of the Na+ channel (amiloride), 3Na-2K adenosinetriphosphatase (acetylstrophanthidin), and Na-K-2Cl cotransporter (furosemide) to anesthetized dogs and/or baboons. Tracheal ciliary beat frequency was measured by using heterodyne laser light scattering. Tracheal mucus velocity (TMV) and bronchial mucociliary clearance (BMC) or lung mucociliary clearance were measured by using radioaerosols and nuclear imaging. Respiratory tract fluid output was collected by using a secretion-collecting endotracheal tube. In six dogs, amiloride aerosol -lung deposition, 96 +/- 11 microg (means +/- SE)- had minimal effect, whereas acetylstrophanthidin aerosol (lung deposition, 71 +/- 9 microg) increased BMC, and furosemide (40 mg iv) markedly increased TMV. In five baboons, TMV increased after iv furosemide administration (2 mg/kg) as well as by aerosol (lung deposition, 20 +/- 3 mg), coincident with increases in ciliary mucus coupling from 11.5 +/- 0. 1 to 29.5 +/- 0.4 and 46.5 +/- 0.7 microm/beat, respectively. Furosemide also increased lung mucociliary clearance in baboons. In dogs, respiratory tract fluid output increased after intravenous furosemide from 2.2 +/- 0.5 to 6.8 +/- 1.7 mg/min. When combined with dry-air inhalation, furosemide failed to stimulate TMV and reversed the inhibition of BMC by dry air. Thus pharmacological manipulation of the Na-K-2Cl cotransporter and the 3Na-2K adenosinetriphosphatase pump may provide increases of clinical relevance in airway hydration and mucociliary transport. PMID- 9338448 TI - Vascular tree structure affects lung blood flow heterogeneity simulated in three dimensions. AB - Pulmonary arterial tree structures related to blood flow heterogeneity were simulated by using a symmetrical, bifurcating model in three-dimensional space. The branch angle (Theta), daughter-parent length ratio (rL), branch rotation angle (phi), and branch fraction of parent flow (gamma) for a single bifurcation were defined and repeated sequentially through 11 generations. With phi fixed at 90 degrees , tree structures were generated with Theta between 60 and 90 degrees , rL between 0.65 and 0.85, and an initial segment length of 5.6 cm and sectioned into 1-cm3 samples for analysis. Blood flow relative dispersions (RD%) between 52 and 42% and fractal dimensions (Ds) between 1.20 and 1.15 in 1-cm3 samples were observed even with equal branch flows. When gamma not equal 0.5, RD% increased, but Ds either decreased with gravity bias of higher branch flows or increased with random assignment of higher flows. Blood flow gradients along gravity and centripetal vectors increased with biased flow assignment of higher flows, and blood flows correlated negatively with distance only when gamma not equal 0.5. Thus a recursive branching vascular tree structure simulated Ds and RD% values for blood flow heterogeneity similar to those observed experimentally in the pulmonary circulation due to differences in the number of terminal arterioles per 1-cm3 sample, but blood flow gradients and a negative correlation of flows with distance required unequal partitioning of blood flows at branch points. PMID- 9338449 TI - Ultrasound Doppler estimates of femoral artery blood flow during dynamic knee extensor exercise in humans. AB - Ultrasound Doppler has been used to measure arterial inflow to a human limb during intermittent static contractions. The technique, however, has neither been thoroughly validated nor used during dynamic exercise. In this study, the inherent problems of the technique have been addressed, and the accuracy was improved by storing the velocity tracings continuously and calculating the flow in relation to the muscle contraction-relaxation phases. The femoral arterial diameter measurements were reproducible with a mean coefficient of variation within the subjects of 1.2 +/- 0.2%. The diameter was the same whether the probe was fixed or repositioned at rest (10.8 +/- 0.2 mm) or measured during dynamic exercise. The blood velocity was sampled over the width of the diameter and the parabolic velocity profile, since sampling in the center resulted in an overestimation by 22.6 +/- 9.1% (P < 0.02). The femoral arterial Doppler blood flow increased linearly (r = 0.997, P < 0.001) with increasing load [Doppler blood flow = 0.080 . load (W) + 1.446 l/min] and was correlated positively with simultaneous thermodilution venous outflow measurements (r = 0.996, P < 0.001). The two techniques were linearly related (Doppler = thermodilution . 0.985 + 0.071 l/min; r = 0.996, P < 0.001), with a coefficient of variation of approximately 6% for both methods. PMID- 9338447 TI - Roles of hydration, sodium, and chloride in regulation of canine mucociliary transport system. AB - To gain insight into the homeostatic mechanisms regulating airway ion/water fluxes and mucociliary transport, the canine tracheobronchial airway fluid was perturbed by deposition of hypo- and hyperosmotic aerosols for >1 h. Tracheal ciliary beat frequency (CBF) was measured by using heterodyne laser light scattering. Tracheal mucus velocity (TMV) and bronchial mucociliary clearance (BMC) were measured by using radioaerosols and nuclear imaging. Respiratory tract fluid output (RTFO) was collected by using a secretion-collecting endotracheal tube. In six dogs, CBF increased during water deposition in the airways to 180 +/ 30 mg/min and RTFO increased from 2.2 +/- 0.5 to 18.3 +/- 1.6 mg/min, accounting for <10% of the fluid deposition. TMV and BMC were unchanged. CBF, TMV, and BMC were markedly increased by inhalation of aerosolized 3.4 M NaCl. Aerosolized 0.85 M NaCl, in contrast, decreased BMC. In this case, RTFO represented 24% of aerosol deposition. Aerosolized 0.85 M choline chloride and 0.85 M sodium gluconate enhanced BMC and TMV concurrent with a decrease in CBF. RTFO of sodium gluconate studies exceeded 50% of aerosol deposition. Thus the airways appear to have transepithelial compensatory mechanisms that reduce the impact of a moderate increases in NaCl and hydration load, but when these responses cannot adequately respond because of the delivery of impermeable ions or very high tonicity, removal of the challenges are affected by a stimulation of mucociliary transport. PMID- 9338450 TI - Analysis of myosin heavy chain mRNA expression by RT-PCR. AB - An assay was developed for rapid and sensitive analysis of myosin heavy chain (MHC) mRNA expression in rodent skeletal muscle. Only 2 microg of total RNA were necessary for the simultaneous analysis of relative mRNA expression of six different MHC genes. We designed synthetic DNA fragments as internal standards, which contained the relevant primer sequences for the adult MHC mRNAs type I, IIa, IIx, IIb as well as the embryonic and neonatal MHC mRNAs. A known amount of the synthetic fragment was added to each polymerase chain reaction (PCR) and yielded a product of different size than the amplified MHC mRNA fragment. The ratio of amplified MHC fragment to synthetic fragment allowed us to calculate percentages of the gene expression of the different MHC genes in a given muscle sample. Comparison with the traditional Northern blot analysis demonstrated that our reverse transcriptase-PCR-based assay was reliable, fast, and quantitative over a wide range of relative MHC mRNA expression in a spectrum of adult and neonatal rat skeletal muscles. Furthermore, the high sensitivity of the assay made it very useful when only small quantities of tissue were available. Statistical analysis of the signals for each MHC isoform across the analyzed samples showed a highly significant correlation between the PCR and the Northern signals as Pearson correlation coefficients ranged between 0.77 and 0.96 (P < 0.005). This assay has potential use in analyzing small muscle samples such as biopsies and samples from pre- and/or neonatal stages of development. PMID- 9338451 TI - Red cell distortion and conceptual basis of diffusing capacity estimates: finite element analysis. AB - To understand the effects of dynamic shape distortion of red blood cells (RBCs) as it develops under high-flow conditions on the standard physiological and morphometric methods of estimating pulmonary diffusing capacity, we computed the uptake of CO across a two-dimensional geometric capillary model containing a variable number of equally spaced RBCs. RBCs are circular or parachute shaped, with the same perimeter length. Total CO diffusing capacity (DLCO) and membrane diffusing capacity (DMCO) were calculated by a finite element method. DLCO calculated at two levels of alveolar PO2 were used to estimate DMCO by the Roughton-Forster (RF) technique. The same capillary model was subjected to morphometric analysis by the random linear intercept method to obtain morphometric estimates of DMCO. Results show that shape distortion of RBCs significantly reduces capillary diffusive gas uptake. Shape distortion exaggerates the conceptual errors inherent in the RF technique (J. Appl. Physiol. 79: 1039-1047, 1995); errors are exaggerated at a high capillary hematocrit. Shape distortion also introduces additional error in morphometric estimates of DMCO caused by a biased sampling distribution of random linear intercepts; errors are exaggerated at a low capillary hematocrit. PMID- 9338452 TI - Respiratory muscle reserve in rats during heavy exercise. AB - The extent to which the respiratory pump muscles limit maximal aerobic capacity in quadrupeds is not entirely clear. To examine the effect of reduced respiratory muscle reserve on aerobic capacity, whole body peak oxygen consumption (VO2 peak) was measured in healthy Sprague-Dawley rats before and after Sham, unilateral, or bilateral hemidiaphragm denervation (Dnv) surgery. VO2 peak was determined by using a graded treadmill running test. Hemidiaphragm paralysis was verified after testing by recording the absence of electromyographic activity during inspiration. Before surgery, VO2 peak averaged 86, 87, and 92 ml . kg-1 . min-1 for the Sham, unilateral, and bilateral Dnv groups, respectively. Two weeks after surgery, there was no significant change in VO2 peak for either the Sham or unilateral Dnv group. However, VO2 peak decreased approximately 19% in the bilateral Dnv group 2 wk after surgery. These findings strongly suggest that the pulmonary system in rats is designed such that during heavy exercise, the remaining respiratory pump muscles are able to compensate for the loss of one hemidiaphragm, but not of both. PMID- 9338453 TI - Self-reported use of mammography among women aged > or = 40 years -- United States, 1989 and 1995. AB - In 1997, breast cancer will be diagnosed in an estimated 180,200 women, and 43,900 women will die from the disease (1). Early detection combined with timely and appropriate treatment can alter the progress of and reduce mortality from this disease (2). Effective screening procedures are available to detect breast cancer in its early stages. However, the benefits of breast cancer screening to reduce mortality in the population can be achieved only if screening guidelines are followed and a large proportion of women receive screening examinations regularly. To estimate the state-specific proportions of women aged > or =40 years who reported receiving a mammogram during the preceding 2 years, CDC analyzed data from the Behavioral Risk Factor Surveillance System (BRFSS) for 1989 and 1995. This report presents the findings, which indicate that, from 1989 to 1995, the percentage of women aged > or =40 years who reported receiving a mammogram during the preceding 2 years increased in all 39 states in the survey. PMID- 9338454 TI - Mortality patterns -- preliminary data, United States, 1996. AB - In 1996, the estimated number of deaths in the United States totaled 2,322,265, slightly higher than the final total in 1995 of 2,312,132. Despite this slight increase in the total number of deaths in 1996, the preliminary crude death rate (875.4 per 100,000 population) declined slightly from that in 1995 and was equal to the crude death rate in 1994; in addition, in 1996 the age-adjusted death rate (493.6 per 100,000 U.S. standard population) reached an all-time low. This report summarizes preliminary 1996 vital statistics data from CDC's National Center for Health Statistics (NCHS) (1) and compares these data with data from 1995 and, for life expectancy, data from 1900 to 1995. The findings indicate declines in the rates for most leading causes of death and the infant mortality rate and an increase in overall estimated life expectancy. PMID- 9338455 TI - Outbreak of invasive group A Streptococcus associated with varicella in a childcare center -- Boston, Massachusetts, 1997. AB - Group A Streptococcus (GAS) causes common childhood diseases such as streptococcal pharyngitis and impetigo and can cause severe, life-threatening invasive disease including streptococcal toxic-shock syndrome and necrotizing fasciitis. Invasive GAS disease occurs when GAS infects a normally sterile site. Clusters of invasive GAS infection previously had not been reported among children in school or childcare centers (CCCs). However, on February 13, 1997, the Boston Public Health Commission received reports of cases of concurrent invasive GAS and varicella infection among two of 14 children in the same CCC classroom. Because of the potential for further spread of invasive disease, the Boston Public Health Commission initiated an investigation of these cases. This report describes the findings of the investigation, including risk factors for infection, and recommended prevention measures. The findings indicate the potential for widespread GAS infection and carriage in CCCs and suggest that, in this outbreak, antecedent use of varicella vaccine would have prevented cases of invasive GAS. PMID- 9338456 TI - Hantavirus pulmonary syndrome -- Chile, 1997. AB - Hantavirus pulmonary syndrome (HPS) was first recognized in Chile in October 1995; through July 1997, nine cases had been identified in the country. However, during August 1-October 8, 1997, a total of 12 persons with HPS, including two family clusters, were recognized. A collaborative investigation is under way to determine the magnitude of this outbreak, the associated rodent reservoir for hantaviruses, and the major risk factors for human infection. This report summarizes the preliminary results of the ongoing investigation, which suggest that the outbreak among humans is paralleled by exceptionally high densities of potential rodent reservoir species. PMID- 9338457 TI - Results of the public health response to Pfiesteria workshop -- Atlanta, Georgia, September 29-30, 1997. AB - On September 29-30, 1997, CDC sponsored a workshop to coordinate a multistate response to public health issues about Pfiesteria piscicida. Workshop attendees included representatives from the health departments of eight states (Delaware, Florida, Georgia, Maryland, North Carolina, South Carolina, Virginia, and West Virginia) and the District of Columbia, the U.S. Food and Drug Administration, the National Institutes of Health's National Institute of Environmental Health Sciences, CDC's National Institute for Occupational Safety and Health, and the U.S. Environmental Protection Agency. PMID- 9338458 TI - Prognostic factor development: an important caution from a small study. PMID- 9338459 TI - Primary small cell carcinoma of the esophagus: a review of the literature with emphasis on therapy and prognosis. AB - BACKGROUND: Few studies of patients with esophageal small cell carcinoma (SCC) have been conducted. Choice of treatment remains controversial. METHODS: The authors analyzed data on 199 evaluable esophageal SCC patients, selected from among 230 patients found in the literature, and a data extraction form that recorded 11 features was completed. To allow for the evaluation of prognostic factors that influenced survival, the patients were grouped according to limited stage (LS), which was defined as disease confined to the esophagus, or extensive stage (ES), which was defined as disease that had spread beyond locoregional boundaries. Univariate and multivariate analyses were performed. Treatment was categorized as either local or local with systemic; for the ES cases, the categories were defined as treatment versus no treatment. RESULTS: The tumor site was described in 178 cases (89%). Mean tumor size was 6.1. Pure SCC was found in 137 cases (68.8%), whereas 62 cases (31.2%) showed mixed SCC; 93 (46.7%) were LS, whereas 95 (47.7%) were ES. In 11 cases (5.5%), the stage was not determined. There was a significant difference in survival between patients with LS and those with ES (P < 0.0001). The median survival was 8 months for patients with LS and 3 months for those with ES. Univariate analysis of LS showed 3 significant prognostic factors: age (for patients age < or =60 years, the median survival was 11 months, whereas for those age >60 years, the median survival was 6 months), tumor size (for those with tumors < or =5 cm, the median survival was 12 months, whereas for those with tumors >5 cm, the median survival was 4 months), and type of treatment (with local plus systemic treatment, the median survival was 20 months, whereas with local it was 5 months). In multivariate analysis, tumor size (P = 0.007) and type of treatment (P < 0.001) were shown to be independent predictive variables. CONCLUSIONS: Esophageal SCC is an aggressive type of tumor. This study shows that there are significant differences between LS and ES and that in LS, both tumor size and type of treatment are possible prognostic factors. PMID- 9338460 TI - Malignant mucosal melanoma of the head and neck: review of the literature and report of 14 patients. AB - BACKGROUND: Fortunately, primary malignant mucosal melanoma of the head and neck is a rare entity. A paucity of data elucidating the predictive factors as well as the unpredictable and aggressive biologic behavior of mucosal melanoma compound the vexing clinical situation. This review summarizes what the literature reveals about the epidemiology, patient survival, patterns of local recurrence, and local and distant metastasis of the disease. Over 1000 patients with this disease have been reported. Survivals at 5 and 10 years is 17% and 5%, respectively. Approximately 19% of patients present with lymph node metastasis and another 16% develop lymph node metastases after treatment, whereas 10% present with distant metastasis. Local metastasis does not affect survival; this is in sharp contrast with skin melanoma. Over 50% of patients experience local treatment failure, and salvage treatment is effective in only 25% of these cases. Local failure is the harbinger of distant metastases. Patients with nasal mucosal melanoma have a 31% 5-year survival rate, whereas sinus melanoma patients fare poorly, with a 0% rate of 5-year survival. METHODS: The authors conducted a retrospective review of 14 patients with characteristics similar to those in the literature in terms of outcome. RESULTS: The 5-year survival rate for these patients was 14%. Whole-body positron emission tomography was performed on 3 patients to detect metastatic disease. The patterns of local recurrence, distant metastasis, and survival for these patients were compared with the same data for patients described in the literature. CONCLUSIONS: Surgery appears to have the greatest efficacy in the management of mucosal melanoma, although radiation therapy may play an increasingly important role in the future. PMID- 9338462 TI - Predictive value of genetic diagnosis for cancer micrometastasis: histologic and experimental appraisal. AB - BACKGROUND: The recently introduced genetic diagnosis of cancer micrometastasis is quite attractive because of its high detection sensitivity. Not infrequently, however, there are marked discrepancies between genetic and conventional histologic diagnoses, especially concerning lymph nodes from colon carcinoma patients. Because the histologic approach has long been relied on in the clinic, the reasons for the differences in results need to be elucidated. METHODS: Serial sections of genetic diagnosis positive but histologic negative lymph nodes of colon carcinoma patients were prepared for histologic and immunohistochemical studies. To investigate the possible contaminating influence of DNA sequences derived from degraded carcinoma cells from the primary site through the lymphatics, the authors also injected purified DNA of human colon carcinoma cells (SW480) into the foot pads of rats and sequentially examined lymph nodes using genetic diagnosis methodology. RESULTS: Careful histologic examination of genetic diagnosis positive, histologically negative lymph nodes of colon carcinoma patients confirmed the absence of cancer cells, whereas p53 protein was immunohistochemically demonstrated to be present in the cytoplasm of sinus histiocytes. In the rat experiment, positive reactions were obtained with the inguinal lymph nodes beginning 30 minutes after the injection, and lymph nodes at various sites were subsequently affected, even after a 72-hour period. CONCLUSIONS: The current study thus suggests that positive results with genetic diagnosis may simply indicate the presence of tumor DNA and do not necessarily mean that viable cancer cells are present. Although the genetic approach may still hold promise for the detection of cancer micrometastases, its predictive value should be carefully assessed clinicopathologically, because its supersensitivity may be associated with a greatly increased false-positive rate. PMID- 9338461 TI - Is chemoradiation feasible in elderly patients? A study of 17 patients with anorectal carcinoma. AB - BACKGROUND: Cancer in the elderly is becoming an increasing public health problem. Nevertheless, several authors have noted the relative lack of information regarding the treatment of cancer in the elderly. The aim of this study was to determine the tolerance of concomitant chemoradiation in patients age > or = 75 years with anorectal carcinoma. METHODS: The patients were selected for treatment on the basis of the absence of major concurrent diseases, normal blood count values, good cardiac and renal function, and good general condition (defined as not requiring personal assistance). Seventeen patients (8 men and 9 women with a median age of 79 years [range, 75-90 years]) were treated with concomitant chemoradiation (bolus mitomycin C, 10 mg/m2 on Day 1 and continuous infusion 5-fluorouracil [5-FU], 1000 mg/m2 for 24 hours on Days 1-4 [FUMIR]). The doses and volumes of pelvic radiation therapy ranged between 38-45 grays according to the primary tumor site and the intent of treatment (curative vs. palliative). RESULTS: The total incidence of Radiation Therapy Oncology Group Grade 3 acute toxicity was 18% (3 of 17 patients). Only 1 patient (6%) was unable to complete the treatment course. With a median follow-up of 26 months, no severe late toxicity was recorded. Sixteen of 17 had >50% reduction in the greatest dimension of the lesion, 6 patients had a complete response (2 rectal and 4 anal tumors), and 12 patients preserved their sphincter function. Of the four patients who had presented with pelvic pain, all had pain relief. Of the six patients who had presented with rectal bleeding, the bleeding was controlled in five patients. CONCLUSIONS: Concomitant chemoradiation according to the FUMIR schedule used in selected patients age > or = 75 years with anorectal carcinoma can be performed safely. PMID- 9338463 TI - The impact of surgical adjuvant thoracic radiation therapy for patients with nonsmall cell lung carcinoma with ipsilateral mediastinal lymph node involvement. AB - BACKGROUND: Previous nonsmall cell lung carcinoma studies have shown that patients with ipsilateral mediastinal (N2) lymph node involvement who underwent surgical resection have a greater local recurrence rate than those with less lymph node involvement (N0, N1). Therefore, it was hypothesized that complete surgical clearance of subclinical lymph node disease is difficult in N2 patients and that adjuvant postoperative thoracic radiotherapy (TRT) may be beneficial. METHODS: A retrospective review was performed to determine the local recurrence and survival rates for patients with N2 disease undergoing complete surgical resection with or without adjuvant TRT. Between 1987 and 1993 at the Mayo Clinic, 224 patients underwent complete resection of N2 nonsmall cell lung carcinoma. More than one mediastinal lymph node station was sampled in 98% of patients; 39% then received adjuvant TRT (median dose, 50.4 grays). RESULTS: The median follow up time was 3.5 years for the patients who were alive at the time of the analysis. The surgery alone versus surgery plus TRT groups were well balanced with respect to gender, age, histology, tumor grade, number of mediastinal lymph node stations dissected or involved, and involved N1 lymph node number. There were slightly more patients with right lower lobe lesions (compared with other lobes), patients with multiple lobe involvement, and patients with only one N2 lymph node involved in the surgery alone group. After treatment with surgery alone, the actuarial 4-year local recurrence rate was 60%, compared with 17% for treatment with adjuvant TRT (P < 0.0001). The actuarial 4-year survival rate was 22% for treatment with surgery alone, compared with 43% for treatment with adjuvant TRT (P = 0.005). On multivariate analysis, the addition of TRT (P = 0.0001), absence of superior mediastinal lymph node involvement (P = 0.005), and fewer N1 lymph nodes involved (P = 0.02) were independently associated with improved survival rate. CONCLUSIONS: This study, which to the authors' knowledge is the largest evaluating adjuvant TRT in N2 nonsmall cell lung carcinoma, suggests that adjuvant TRT may improve local control and survival. PMID- 9338465 TI - Outcomes of patients with local recurrence of cutaneous malignant melanoma: a population-based study. AB - BACKGROUND: The definition of local recurrence of cutaneous malignant melanoma varies. The outcomes of patients with a local recurrence reported in the literature also vary, but the appearance of a local recurrence has generally been considered a negative prognostic sign. Few studies have been population-based thus far. METHODS: During the period 1976-1997, 3706 patients with cutaneous malignant melanoma (including 575 patients with melanoma in situ) were registered in a population-based regional cancer registry. Local recurrence was defined as a recurrence within the scar or transplant with no signs of regional or distant spread of the disease. Prognostic factors were investigated using univariate and multivariate analytic techniques. The prognostic importance of a local recurrence in terms of survival was analyzed using the Cox proportional hazards regression model, with local recurrence as a time-dependent covariate. RESULTS: Local recurrence as a first event was rare (occurring in 48 of 3706 patients, or 1.3%). Twenty-eight percent (11 of 39) of the patients with local recurrence of invasive primary melanoma developed distant metastases and subsequently died. Only ulceration had prognostic significance in univariate analysis. A Cox analysis, with melanoma death as the endpoint and local recurrence as a time-dependent covariate, demonstrated a relative risk of 1.3 associated with local recurrence; however, this was not statistically significant (confidence interval, 0.7-2.3). CONCLUSIONS: In this population-based study, local recurrence was a rare event. The outcomes after diagnosis were relatively favorable. The results did not indicate a major detrimental effect on survival from the local recurrence per se. PMID- 9338466 TI - Predicting ten-year survival of patients with primary cutaneous melanoma: corroboration of a prognostic model. AB - BACKGROUND: Recently, the Pigmented Lesion Group at the University of Pennsylvania described a 4-variable model for predicting 10-year survival for patients with primary cutaneous melanoma. The variables are tumor thickness, anatomic site of the lesion, age, and gender. The objective of the current study was to test the validity of this model, employing the large data base of the New York University Melanoma Cooperative Group. METHODS: The predicted probabilities of 10-year survival for 780 patients with primary cutaneous melanoma were determined by multivariate logistic regression, using the 4 variables. RESULTS: The overall 10-year survival rate of the current study group was 78.4%. Of the four variables, tumor thickness, anatomic site of the lesion, and age were found to be independent predictors of survival. Although survival was better for women, gender was not a statistically significant factor in predicting 10-year survival when entered into the multivariate logistic regression model. In the current study, the probability of 10-year survival of patients with melanomas < 0.76 mm ranged from 93-99%, depending on the age and primary site. Age and site had more impact on the prognosis of intermediate and thick melanomas than on thin melanomas. Thus, for melanomas 0.76-1.69 mm, 1.70-3.60 mm, and thicker than 3.60 mm, the probabilities of survival ranged from 70-94%, 39-82%, and 23-68%, respectively. CONCLUSIONS: The wider ranges in survival rates for thicker melanomas, depending on the other variables, emphasize the importance of including variables in addition to tumor thickness in a prognostic model. Using a large data base from a medical center, the current study supports the prognostic multivariate model of the Pigmented Lesions Group of the University of Pennsylvania; however, the authors of the current study did not find gender to be statistically significant in this multivariate model. PMID- 9338464 TI - T-cell lymphocytosis associated with lymphocyte-rich thymoma. AB - BACKGROUND: Peripheral T-cell lymphocytosis is found on very rare occasions in patients with thymomas. The immunophenotypic features and clonality of the lymphocytes in tumor and peripheral blood now are elucidating this enigmatic phenomenon. METHODS: The author presents what is believed to be the seventh case of peripheral T-cell lymphocytosis associated with thymoma and reviews the previous six cases. The pathology slides of the thymoma were reviewed with a pathologist who confirmed the presence of neoplastic thymic epithelium with cytokeratin stains. Immunophenotyping by flow cytometry was performed at Dianon Systems, Inc., on both the thymoma cells and cells in the peripheral blood. In addition, gene rearrangement analysis was performed on the peripheral lymphocytes using the previously described Southern blot analysis technique with immunoglobulin probes (heavy chain, kappa light chain, and lambda light chain) and T-cell receptor gene probes (beta and gamma chains). RESULTS: Analyses of the T cells within the thymoma and the peripheral blood confirm that the peripheral T cells are both polyclonal and more mature than those populating the thymoma. Clearly the peripheral T cells are not themselves neoplastic, and yet they represent more than physical "spillover" of the immature tumor-related T cells. CONCLUSIONS: Peripheral T-cell lymphocytosis occurs rarely with locally aggressive, lymphocytic thymomas. Although it is clear that these cells are not neoplastic, as they are in other T-cell proliferative disorders, the etiology of this unusual phenomenon remains obscure and may reflect the perturbation of systemic immunoregulation that accompanies thymic neoplasia. It is important to differentiate this condition from T-cell lymphomas or leukemia to treat affected patients appropriately. PMID- 9338467 TI - Breast carcinoma stage in relation to time interval since last mammography: a registry-based study. The Romagna Cancer Registry and Collaborators. AB - BACKGROUND: In the Romagna Region of Italy, mammography screening for breast carcinoma (BC) was implemented as a routine practice in the regular healthcare system. The Romagna Cancer Registry evaluated the effects of self-selection for mammography on the stage of BC at the time of presentation. METHODS: Of the 851 invasive BC cases registered in 1989-1991, tumor size (T) was documented for 790 (93%) and lymph node status (N) for 681 (80%). Mammography experience was determined by cross-checks with the radiology files in the area. The Mantel Haenszel chi-square test stratified by age was used to evaluate the difference in the proportion of advanced BC between subsets of patients diagnosed at increasing time intervals since their most recent mammography and those with no previous examination. RESULTS: The incidence of T2+/N1+ (T2 or worse and/or lymph node positive) BC was 71% among patients without previous mammography. No advantage was observed for patients diagnosed within 6-12 months of their last mammography (69%). A marked advantage was associated with intervals varying between 12-23 months (45%; P < 0.001) and between 24-35 months (43%; P < 0.001). For longer intervals, the proportion of T2+/N1+ disease stabilized at 56%. For patients with any interval > 5 years (range, 5-22 years; median, 11 years) the relative stage advantage was significant (P = 0.013). CONCLUSIONS: The stage pattern of patients diagnosed with BC 5-22 years after their last mammography suggests that even if health-aware women who volunteered for screening stop undergoing regular mammography, they present for a relatively prompt diagnosis in the event they develop BC. PMID- 9338468 TI - Use of calcium channel blockers and breast carcinoma risk in postmenopausal women. AB - BACKGROUND: The use of calcium channel blockers in an elderly population recently was reported to be associated with the incidence of cancer. The Cardiovascular Health Study, a multisite observational cohort study, provided the opportunity to investigate the epidemiologic association between the use of calcium channel blockers and breast carcinoma risk in 3198 women age > or = 65 years. METHODS: Standard questionnaires and clinical procedures were administered at four study sites annually from 1989-1990 to 1993-1994. Drug usage was assessed by a medication inventory and hospitalizations for 75 incident invasive breast carcinoma cases were identified using International Classification of Diseases-9 Clinical Modification codes. Time-dependent Cox proportional hazards regression models were used to assess associations between incident breast carcinoma and the use of specific antihypertensive medication including calcium channel blockers. RESULTS: In adjusted Cox proportional hazards models, an elevated risk of breast carcinoma was associated with use of calcium channel blockers (hazard ratio [HR]: 2.57; 95% confidence interval [CI], 1.47-4.49). This association persisted when the comparison group was users of other antihypertensive medication. No associations between the use of other antihypertensive medication with incident breast carcinoma were found. Associations were enhanced by assessment of high dose at baseline (HR: 4.42; 95% CI, 1.37-14.27) and when calcium channel blockers were combined with estrogen use (HR: 4.48; 95% CI, 1.58-12.75). The association was found to be strongest for the use of estrogens with immediate release calcium channel blockers (HR: 8.48; 95% CI, 2.99-24.08). CONCLUSIONS: Although the number of cases was limited in this observational study, associations found between the use of calcium channel blockers and incident invasive breast carcinoma warrant further investigation. Site specific carcinomas should be included as an outcome of ongoing and planned long term clinical trials using calcium channel blockers. PMID- 9338469 TI - Alpha-1 acid glycoprotein is an immunosuppressive factor found in ascites from ovaria carcinoma. AB - BACKGROUND: Although the ascites of patients with ovarian carcinoma has been reported to contain immunosuppressive factors, the identity and source of this activity has not been fully elucidated. The objective of this study was to describe the purification of a single immunosuppressive protein, alpha-1 acid glycoprotein, from ovarian carcinoma ascites, identify its site of production, and describe a possible mechanism by which it inhibits lymphocytes. METHODS: Ascites from proteins from five patients with epithelial ovarian carcinoma first were differentially precipitated by size with different concentrations of polyethylene glycol and then separated on the basis of isoelectric focusing. The protein factions then were placed in a lymphocyte proliferation assay to determine immunosuppressive activity. Western blot analysis was used to identify alpha-1 acid glycoprotein as an immunosuppressive protein in ascites. Total RNA was extracted from ovarian and hepatic cell lines as well as primary and recurrent ovarian tumor samples. Reverse-transcriptase polymerase chain reaction then was utilized to identify the site of production of this protein. Purified alpha-1 acid glycoprotein was placed in lymphocyte culture and its effects on lymphocyte interleukin-2 (IL-2) production were measured by enzyme-linked immunoadsorbent assay. RESULTS: Addition of purified alpha-1 acid glycoprotein to the lymphocyte assay resulted in a 60% decrease in lymphocyte proliferation (P < 0.05). Alpha-1 acid glycoprotein transcript was not identified in ovarian tumor cells. The addition of purified alpha-1 acid glycoprotein to the lymphocyte culture resulted in a 65% decrease in IL-2 secretion into the media (P < 0.05). CONCLUSIONS: Alpha-1 acid glycoprotein is an immunosuppressive protein purified from ovarian carcinoma ascites. It is not expressed primarily by ovarian carcinoma cells. It appears to inhibit IL-2 secretion by lymphocytes. PMID- 9338470 TI - Production of extracellular matrix-degrading proteinases by primary cultures of human epithelial ovarian carcinoma cells. AB - BACKGROUND: The authors analyzed the secretion of extracellular matrix-degrading proteinases, including urinary-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2, gelatinase A), and MMP-9 (gelatinase B), by short term primary cultures of epithelial ovarian carcinoma cells derived from primary ovarian tumors, intraperitoneal metastases, or ascites. The presence of these enzymatic activities in samples of ascites was also evaluated. The effect of adhesive substratum on proteinase production was determined. METHODS: A coupled spectrophotometric assay was utilized to evaluate the initial rate of plasminogen activation by u-PA in conditioned medium; this involved monitoring the activity of generated plasmin with a colorimetric substrate. MMP activity was evaluated by gelatin zymography. RESULTS: Ascitic fluids from 18 patients contained u-PA, MMP 2, and MMP-9. However, short term primary cultures of cells derived from primary ovarian tumors (OVET), metastatic lesions (OVEM), or ascites (OVEA) produced very low levels of u-PA. Production of u-PA by OVET and OVEM cells was regulated by adhesive substratum. Conditioned media from OVET, OVEM, and OVEA cells contained high levels of both MMP-2 and MMP-9. MMP-9 levels decreased with increasing passage in culture, whereas MMP-2 activity was maintained. Production of neither MMP-2 nor MMP-9 was regulated by adhesive substratum. CONCLUSIONS: These results demonstrate that primary cultures of epithelial ovarian carcinoma cells derived from three distinct anatomic locations produce MMP-2 and MMP-9, with low level secretion of u-PA. These data suggest that MMPs, particularly MMP-2, may play a significant role in the intraperitoneal invasion of ovarian carcinoma cells. PMID- 9338471 TI - Combined treatment of invasive bladder carcinoma with transurethral resection, induction chemotherapy, and radical radiotherapy plus concomitant protracted infusion of cisplatin and 5-fluorouracil: a phase I study. AB - BACKGROUND: The aim of this study was to define the maximum tolerated doses (MTDs) of cisplatin (CDDP) and 5-fluorouracil (5-FU) administered as protracted intravenous infusion (PVI) during hyperfractionated radiotherapy (HFRT) administered with organ-sparing intent to patients with infiltrating transitional cell carcinoma of the bladder (TCCB). METHODS: Twenty-five patients with T2 T4aNXM0 TCCB were enrolled in this study. After a complete transurethral resection, bladder mapping, and two cycles of induction chemotherapy, patients were submitted to HFRT and CDDP + 5-FU as concomitant PVI at escalating dose levels until MTDs were reached. Treatment efficacy was also evaluated, in terms of complete response (CR) rates and cystectomy free, disease free, and overall survival. RESULTS: Combined treatment was well tolerated. The recommended doses for Phase II studies of PVI chemotherapy and radiotherapy for patients with invasive bladder carcinoma are CDDP 5 mg/m2/day and 5-FU 220 mg/m2/day. Twenty four patients were evaluable for response: 21 (87.5%) had CR and 3 PR. After a median follow-up of 31 months (range, 11-49 months), 18 of 21 patients with CRs (86%) were alive: 15 (71.4%) had tumor free bladder, of whom 3 had superficial recurrence successfully treated with endovesical therapy and 1 had distant metastases. Three patients were submitted to cystectomy, one for superficial recurrence and hematuria and two for invasive bladder recurrence. CONCLUSIONS: This study defines the MTDs of CDDP and 5-FU concomitantly administered with hyperfractionated radiotherapy. The low toxicity observed and the high CRs and bladder preservation rates deserve further study. PMID- 9338473 TI - Socioeconomic status and comorbidity among newly diagnosed cancer patients. AB - BACKGROUND: Many studies found better cancer survival in patients with a high socioeconomic status (SES) than in patients with a low SES. Comorbidity at the time of diagnosis may be more frequent in patients of lower SES, and negatively influences their survival. The authors studied the association between SES and serious comorbidity at the time of diagnosis among newly diagnosed cancer patients in The Netherlands. METHODS: Included in the analyses were patients registered in 1993 in the population-based Eindhoven Cancer Registry (southeastern Netherlands) with one of the most common carcinomas: breast (n = 457), lung (n = 442), colorectum (n = 384), prostate (n = 240), and stomach (n = 118). Information regarding comorbidity came from medical records. The SES of the patients was derived from their postal code of residence and stratified into three categories. The risk of being diagnosed with at least one other chronic condition was calculated using logistic regression analyses. RESULTS: The risk of being diagnosed with at least one other chronic condition was higher among patients with a low or intermediate SES than among those with a high SES for the five sites combined as well as for carcinomas of the breast or lung. The gradient was less clear for patients with colorectal carcinoma, whereas no socioeconomic variation in comorbidity was found for patients with carcinomas of the prostate or stomach. CONCLUSIONS: Socioeconomic variation in the prevalence of serious comorbidity at the time of diagnosis does exist in some cancer sites, which may explain (partly) the socioeconomic gradient in survival observed in patients with tumors in these sites. PMID- 9338472 TI - The prognostic value of p53 nuclear overexpression and MIB-1 as a proliferative marker in transitional cell carcinoma of the bladder. AB - BACKGROUND: There is controversy regarding the value of biologic markers as prognostic indicators independent of clinicopathologic parameters in transitional cell carcinoma (TCC) of the bladder. The authors examined the prognostic value of p53 tumor suppressor gene expression and the proliferative marker MIB-1 in TCC of the bladder. METHODS: Fresh frozen samples from 114 TCCs of the bladder and 13 normal bladders were studied by immunohistochemistry, using monoclonal antibodies MIB-1 for proliferation and PAb 1801 for p53 nuclear overexpression. Scores were determined in each case by counting at least 500 nuclei per slide in 3-5 selected regions. Patients were stratified for both markers into two groups for time-event analysis, according to the median number of nuclei stained. Patients with nuclear staining below the median value of the score were considered negative in the statistical analysis. Quantitative immunostaining was analyzed in relation to the time to recurrence, progression, and cancer death, and compared with clinical and pathologic parameters for prognostic significance in univariate and multivariate analysis (stepwise logistic regression). RESULTS: Median nuclear overexpression of p53 was 22% and that of MIB-1 28%. There was a strong association between proliferation and p53 nuclear overexpression (P < 0.0001). Progression free and disease specific survival rate estimates (log rank test) were significantly lower in patients with p53 or MIB-1 scores above 22% and 28%, respectively. Multivariate analysis indicated that stage as well as p53 and MIB-1 immunostaining provided independent prognostic information. CONCLUSIONS: Quantitative immunohistochemical evaluation of nuclear p53 and MIB-1 immunostaining inexpensively provides prognostic indicators in cases of TCC of the bladder. PMID- 9338474 TI - Polymorphism in the tumor necrosis factor-alpha promotor region and in the heat shock protein 70 genes associated with malignant tumors. AB - BACKGROUND: Tumor necrosis factors (TNFs) and heat shock protein 70 (hsp70) are determining factors in immunologic mechanisms to tumor cells. The authors designed a case-controlled study to investigate the potential association of the polymorphisms of TNF-alpha and of hsp70-2 and hsp70-hom genes with malignant tumors. METHODS: The authors used an allele specific polymerase chain reaction to characterize the variation of the TNF-alpha promotor region in 124 unrelated Tunisian patients with malignant tumors (non-Hodgkin's lymphoma, breast carcinoma, and other tumors) and 106 healthy control subjects. Using polymerase chain reaction and restriction enzyme digestion, polymorphic analysis of hsp70-2 and hsp70-hom genes was performed in patients with non-Hodgkin's lymphoma, in those with breast carcinoma, and in control subjects. RESULTS: Analysis of TNF alpha polymorphism in patients with malignant tumors and in control subjects demonstrated a high relative frequency of the TNF2 allele in the cancer patients. The relative risk (RR) of lymphoma was especially high in association with TNF1/TNF2 heterozygotes (RR = 6.7; P < or = 0.0001). Polymorphism analysis of the hsp70-2 and hsp70-hom genes in patients with lymphoma and in those with breast carcinoma revealed that these patients had highly significant differences in the genotypic distribution of these biallelic loci compared with the control subjects. Homozygosity for one hsp70-2 allele was significantly associated with lymphoma (RR = 18.2; P < or = 0.0001) and with breast carcinoma (RR = 16.3; P < or = 0.001). CONCLUSIONS: Tunisian persons carrying the TNF2 allele may have an increased risk of cancer. In this study, non-Hodgkin's lymphoma and breast carcinoma were significantly associated with polymorphism in hsp70 genes. PMID- 9338475 TI - Primary cardiac lymphoma in immunocompetent patients: diagnostic and therapeutic management. AB - BACKGROUND: Primary cardiac lymphoma (PCL) is extremely rare in immunocompetent patients. Different definition criteria have been employed in published series. Prognosis is poor due to diagnostic delay and relevance of the site of disease. METHODS: Two cases observed at the study institution are reported, with a review of 48 cases published in the literature from 1980 to 1996. Only patients with lymphoma confined to the heart and/or pericardium and those with a single and asymptomatic extracardiac site were considered for analysis. RESULTS: Eight patients had minimal extracardiac disease. The most common presentation was unresponsive heart failure. Electrocardiography findings were not specific. PCL usually arose in the right chambers as a mass, with or without pericardial effusion (> 80%). Chest X-rays, transthoracic echocardiography, and computed tomography scans are standard in diagnostic workup, but transesophageal echocardiography (TEE) and magnetic resonance imaging (MRI) showed a sensitivity > 90%. Cytology of pericardial effusion was diagnostic in 67% of cases. Thoracotomy was diagnostic in all cases, whereas less invasive procedures had high false-negative rates. Gross resection has no role. Early anthracycline containing chemotherapy appears to improve survival, whereas the role of radiotherapy has not yet been defined. CONCLUSIONS: The diagnosis of PCL should be considered in patients with a cardiac mass and/or unexplained refractory pericardial effusion. Adequate diagnostic workup, including TEE and MRI, allows confirmation of the early suspicion of PCL. In the absence of a diagnostic cytology, an open biopsy may be indicated to avoid treatment delay. There is no evidence that PCL should be treated differently from other bulky aggressive lymphomas arising at other anatomic sites. PMID- 9338476 TI - Thallium-201 scintigraphy for the evaluation of tumor response to preoperative chemotherapy in patients with osteosarcoma. AB - BACKGROUND: The degree of tumor necrosis after preoperative intravenous chemotherapy is likely to be an indicator of prognosis for patients with primary osteosarcoma. This study was undertaken to evaluate the efficacy of thallium-201 (Tl-201) scintigraphy in assessing response to chemotherapy in patients with osteosarcoma and to correlate the findings with the degree of tumor necrosis assessed histologically. METHODS: Twenty-four patients with biopsy-proven osteosarcoma underwent Tl-201 imaging before and immediately after preoperative intravenous chemotherapy. The maximum pixel counts taken over the tumor divided by those taken of a background region yielded a tumor-to-background ratio. The percentage of change in the tumor-to-background ratio before and after chemotherapy, defined as the alteration ratio, was correlated with the percentage of tumor necrosis. The percentages of tumor necrosis were classified into 4 grades: Grade 1, <50%; Grade 2, 50-89%; Grade 3, 90- 99%; Grade 4, 100%. RESULTS: Three patients with Grade 1 histologic response had an alteration ratio of 31% +/ 4 (mean +/- standard deviation). Three patients with Grade 2 response had an alteration ratio of 76% +/- 9. Nine patients with Grade 3 response had an alteration ratio of 84% +/- 11. Nine patients with Grade 4 response had an alteration ratio of 96% +/- 5. These ratios correlated well with the histologic grades (P < 0.001). CONCLUSIONS: The results of this study suggest that Tl-201 scintigraphy is a powerful tool in the evaluation of tumor response to chemotherapy in patients with osteosarcoma. PMID- 9338477 TI - Chest wall rhabdomyosarcoma. AB - BACKGROUND: Rhabdomyosarcoma is the most common soft tissue sarcoma in the pediatric age group. The primary tumor site is an important prognostic determinant. Axial lesions are associated with decreased survival and provide a clinical challenge. METHODS: A retrospective analysis of the authors' institutional experience between 1972 and 1996 was performed. Patients were from a data base of 302 consecutive cases. RESULTS: Fifteen consecutive patients with chest wall rhabdomyosarcoma were identified. The median age was 16 years (range, 6 months-25 years). Median follow-up was 6.6 years (range, 10 months-18.5 years). Nine patients presented with a mass, six with pain, two with respiratory distress, and one with ulnar neuropathy. The median lesion size was 7 cm (range, 3-16 cm). A surgical procedure was the initial therapy for 13 of 15 patients. Fourteen patients received radiation therapy with a median dose of 4400 cGy. All but one were included in institutional-based trials using multiagent chemotherapy. At last follow-up, 10 patients were alive and disease free, with a median survival of 123 months (range, 51-221 months). Seven of ten survivors underwent a complete resection as their initial therapy. There was no surgical mortality, and only two patients had treatment-related complications. Of the five patients who died, two underwent complete resection as their initial therapy. All five patients had invasive tumors. Four were > 10 cm, 3 were of alveolar subtype, and 2 were embryonal. CONCLUSIONS: Complete resection of chest wall rhabdomyosarcoma is recommended. However, survival is possible for patients with microscopically positive surgical margins with the addition of chemotherapy and radiation. PMID- 9338478 TI - The supportive care needs of newly diagnosed cancer patients attending a regional cancer center. AB - BACKGROUND: The objective of this study was to examine the physical and emotional health status, self-perceived problems, and needs of newly diagnosed cancer patients to determine and plan supportive care strategies. METHODS: A cross sectional survey of newly diagnosed cancer patients attending a regional cancer center during a 6-month period was performed. Patients with breast, colorectal, head and neck, lung, and prostate carcinoma as well as nonmelanoma of the skin were selected randomly. Patients were interviewed prior to their first appointment at the clinic. Physical health status was assessed using the Symptom Distress Scale, psychologic health status was assessed with the General Health Questionnaire (GHQ), day-to-day functioning with the Rapid Disability Scale, and social support with the modified Sarason's Social Support Scale. Perceived needs were assessed in a number of ways, including identification of patients' specific social concerns and informational needs, and by asking them to list their current problems or concerns. RESULTS: Of 156 eligible patients, 134 completed the interview. One hundred and twenty-nine patients (96%) reported current symptoms that included fatigue (66%), worried outlook (61%), difficulty sleeping (48%), and pain (42%). Forty-four patients (33%) were identified as psychologically distressed with a GHQ score of > or = 6. One hundred and fourteen patients (85%) had informational needs, 89 (66%) indicated > or = 1 social concerns, and 55 (41%) reported a need for assistance with day-to-day living. CONCLUSIONS: Patients with newly diagnosed cancer commonly report symptoms related to fatigue, pain, and psychologic distress. Other frequently reported issues relate to the need for information and social concerns regarding the patients' ability to take care of their home and maintain family and other relationships. Awareness of these issues is important for planning supportive care interventions for newly diagnosed cancer patients. PMID- 9338479 TI - Politics and addiction: an update on the tobacco settlement. PMID- 9338480 TI - Treatment of chlamydial genital infection. PMID- 9338481 TI - Current issues in the treatment and prophylaxis of Pneumocystis carinii pneumonia in HIV infection. PMID- 9338482 TI - The potential management of resistant infections with non-antibiotics. AB - The antimicrobial activity of synthetic, non-chemotherapeutic compounds, such as the phenothiazine, methylene blue, has been known since the time of Ehrlich (1854 1915). In this context the term 'non-antibiotics' is taken to include a variety of compounds which are employed in the management of pathological conditions of a non-infectious aetiology, but which modify cell permeability and have been shown to exhibit broad-spectrum antimicrobial activity. The antimicrobial properties of compounds such as phenothiazines, as well as those of other neurotropic compounds, have only been investigated sporadically, and their application to management of microbial infections has not been evaluated. A review of the literature, coupled with a number of more recent investigations, suggests that some of these and other membrane-active compounds enhance the activity of conventional antibiotics, eliminate natural resistance to specific antibiotics (reversal of resistance) and exhibit strong activity against multi-drug resistant forms of Mycobacterium tuberculosis. Thus non-antibiotics may have a significant role in the management of certain bacterial infections. PMID- 9338483 TI - The development of a small-scale biofilm model suitable for studying the effects of antibiotics on biofilms of gram-negative bacteria. AB - A method for the study of membrane-associated biofilm populations of Escherichia coli ATCC 8739 using modified Swinnex filter units was developed. Biofilms were established under carbon limitation in a chemically defined simple salts medium. Cells, pressure filtered on to cellulose nitrate membranes in situ, were perfused from the sterile side. Steady-state conditions were attained at which the growth rate of the sessile cells could be demonstrated to be proportional to the flow rate of medium. The antibiotic susceptibility of these biofilms was examined by including ciprofloxacin within the perfusing medium. Susceptibility of the biofilms to ciprofloxacin was found to be affected not only by its concentration, but also by the growth rate of the bacteria. PMID- 9338484 TI - Potentiation of beta-lactams against Pseudomonas aeruginosa strains by Ro 48 1256, a bridged monobactam inhibitor of AmpC beta-lactamases. AB - Ro 48-1256 is a bridged monobactam inhibitor of Class C beta-lactamases, without significant antibacterial activity of its own. It was tested in combination with imipenem, meropenem, piperacillin and ceftazidime against Pseudomonas aeruginosa isolates, mutants and transconjugants. Imipenem was potentiated against all strains where the AmpC enzyme was inducible or derepressed, with its MICs being reduced from 1-2 mg/L to 0.25-0.5 mg/L for most isolates and from 8-16 mg/L to 1 2 mg/L for those lacking OprD (D2 porin). Ro 48-1256 also abolished in-vitro selection of OprD-deficient mutants by imipenem. Ceftazidime and piperacillin were potentiated against strains derepressed for AmpC enzyme, but not against those where the enzyme remained inducible. For over 90% of AmpC-derepressed organisms, the MICs of ceftazidime were reduced to < or = 8 mg/L by Ro 48-1256 (4 mg/L) and those of piperacillin were reduced to < or = 16 mg/L. Meropenem, which is very stable to AmpC, was not potentiated. Ro 48-1256 did not potentiate piperacillin, ceftazidime or carbapenems when resistance was mediated by Class A, B or D enzymes. Tazobactam, tested as control, had opposite behaviour to Ro 48 1256, potentiating beta-lactams when resistance was due to Class A beta lactamases but failing to reverse resistance mediated by AmpC. Ro 48-1256 could be used with imipenem to overcome resistance mediated by loss of OprD, or with ceftazidime or piperacillin to overcome derepression of AmpC. PMID- 9338486 TI - In-vitro activity of spiramycin and metronidazole alone or in combination against clinical isolates from odontogenic abscesses. AB - One hundred and forty-eight isolates of bacteria from 20 intraoral odontogenic abscesses were tested for their susceptibility to spiramycin and metronidazole alone or in combination. All isolates, except Rothia spp. (one), Enterococcus avium (three), Haemophilus parainfluenzae (one) and Staphylococcus aureus (one) were sensitive to spiramycin and/or metronidazole. Among the anaerobes, spiramycin as well as metronidazole showed good antimicrobial activity against species of Prevotella, Eubacterium, Peptostreptococcus, Bacteroides and Porphyromonas. All the aerobes were resistant to metronidazole. A potential synergic effect was found in 17% of the clinical isolates tested, as the MICs of spiramycin and metronidazole for 25 isolates (21 isolates of Eubacterium, two of lactobacilli, one strain of Peptostreptococcus sp. and one Clostridium clostridiiforme) were significantly reduced by the addition of the other antibiotic. PMID- 9338485 TI - Enhancement of Burkholderia cepacia antimicrobial susceptibility by cationic compounds. AB - Infections in cystic fibrosis (CF) due to Burkholderia cepacia are challenging due to their resistance to antibiotics. We explored a new strategy for increasing the permeability of B. cepacia using cationic agents, including amino compounds, to reduce the MICs of standard antibiotics. Twenty-eight B. cepacia isolates from four CF centres in North America and four non-CF B. cepacia were examined by standard microtitre broth dilution methods for susceptibility to a variety of antibiotics in the presence of non-inhibitory concentrations of diaminoacetone (DAA), methylglyoxal bis-guanylhydrazone (MGBH), chlorpromazine (CPZ) and prochlorperazine (PCPZ). The proportion of isolates with greater than four-fold reductions in MIC in the presence of 0.3 mM CPZ or 0.4 mM PCPZ were 90% and 94% for gentamicin, 80% and 83% for tobramycin, 45% and 17% for ceftazidime, and 35% and 17% for amifloxacin. CPZ showed the same degree of reduction in the MIC of azithromycin in 79% strains (MIC50 reduced to 16 from > or = 256 mg/L). Non-CF B. cepacia showed a greater than four-fold reduction in MIC with CPZ for gentamicin, tobramycin and azithromycin and two-fold reduction for ceftazidime. Little or no reduction in MIC was seen with DAA or MGBH for any antibiotic. Addition of magnesium ions to the medium competitively inhibited any MIC reduction effect seen with the cationic agents. CPZ and PCPZ appeared to enhance the permeability of B. cepacia to antibiotics based upon ionic charge characteristics of the antibiotic. No significant differences were seen in outer membrane protein and lipopolysaccharide profiles between the culture treated with CPZ and the respective control culture of strain B. cepacia ATCC 13945. The fluorescent probe 1N-phenylnaphthylamine had no increased access across the outer membrane in the presence of CPZ for B. cepacia ATCC 13945. However, thin-section electron microscopy revealed separation between the outer membrane and the rest of the cytoplasm accompanied by a widening of the periplasmic space. These data provide a rationale for investigating amino compounds as potential permeability increasing agents against B. cepacia. PMID- 9338487 TI - In-vitro susceptibility, tolerance and glycocalyx production in Streptococcus mutans. AB - We studied the presence of high-level resistance to aminoglycosides, penicillin tolerance and glycocalyx production in 160 isolates of Streptococcus mutans. Susceptibility to amoxycillin, cefazolin, imipenem, erythromycin, clindamycin, vancomycin and teicoplanin was also investigated. Of the isolates analysed, 58.8% produced glycocalyx in vitro and 2.5% were penicillin-tolerant. High-level resistance to streptomycin was found in 16.3% of the isolates, but all were sensitive to all other antibiotics tested. We found no significant relationship between glycocalyx production and high-level streptomycin resistance, penicillin tolerance or antibiotic susceptibility, except for a greater susceptibility to clindamycin and vancomycin in isolates that produced glycocalyx. Although our findings reflect the clinically favourable pattern of susceptibility currently found in this species, the appearance in some isolates of resistance, tolerance and glycocalyx production should be investigated because of the risks involved in endocarditis caused by S. mutans. PMID- 9338488 TI - Permeability to carbapenems of Proteus mirabilis mutants selected for resistance to imipenem or other beta-lactams. AB - An imipenem-resistant mutant of Proteus mirabilis lacked a 26 kDa outer membrane protein (OMP). It has previously been postulated that this protein is a porin, but the present mutant, which was cross-resistant to mecillinam but not to other beta-lactams, proved as permeable to carbapenems as its parent. A mecillinam selected mutant had similar cross-resistance yet retained the 26 kDa OMP, confirming that this protein was not important to resistance. In contrast, cefoxitin-selected mutants retained the 26 kDa protein but had diminished expression of major 41 and 44 kDa OMPs and showed reduced uptake of carbapenems, although this promoted resistance only when a carbapenemase was also present. We conclude that the imipenem-selected mutant owed its resistance to some factor other than porin loss, probably to a lesion in penicillin-binding protein 2. PMID- 9338489 TI - Altered denA and anr gene expression in aminoglycoside adaptive resistance in Pseudomonas aeruginosa. AB - Adaptive resistance to aminoglycoside killing and cytoplasmic accumulation occurs in cultures of originally susceptible Pseudomonas aeruginosa following an initial incubation with aminoglycoside. Anaerobiosis has also been reported to reduce bacterial killing and limit cytoplasmic aminoglycoside accumulation. We hypothesized that a common mechanism may facilitate reduced bacterial killing and aminoglycoside accumulation in both cases. Northern blot analysis of P. aeruginosa adaptively resistant to gentamicin demonstrated increased mRNA levels of both denA (nitrite reductase), which facilitates terminal electron acceptance in the anaerobic respiratory pathway, and its regulatory protein, ANR, in the absence of promoter DNA sequence changes, when compared with controls. These observations suggested that P. aeruginosa may regulate the expression of genes in its anaerobic respiratory pathway in response to aminoglycosides and may explain, at least partially, P. aeruginosa adaptive resistance to aminoglycosides. PMID- 9338490 TI - Molecular genetic analysis of high-level gentamicin resistance in Enterococcus hirae. AB - High-level resistance to gentamicin was studied in seven clinical isolates of Enterococcus hirae. In common with other members of the genus Enterococcus, such resistance in E. hirae was associated with single, large, conjugative plasmids. Molecular genetic analysis revealed five distinct plasmid types amongst the seven isolates. The determinant mediating high-level gentamicin resistance in E. hirae was also shown to be homologous to that already characterized for other enterococcal species. PMID- 9338491 TI - Antibacterial activity of quinolones against coagulase-negative staphylococci and the quinolone resistance-determining region of the gyrA genes from six species. AB - Antibacterial activity of quinolones against three species of coagulase-negative staphylococci was investigated. Tosufloxacin and sparfloxacin exhibited potent activities against Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus saprophyticus compared with other quinolones tested. From the analysis of the DNA sequence in the quinolone resistance-determining region (QRDR), greater than 80% homology was recognized in coagulase-negative staphylococci. A series residue was conserved in all six species at the position corresponding to position 84 in Staphylococcus aureus. PMID- 9338492 TI - Increasing antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in Finland. AB - Respiratory and otitis isolates of 807 Streptococcus pneumoniae, 816 Haemophilus influenzae and 446 Moraxella catarrhalis were collected from 21 clinical microbiology laboratories for antimicrobial susceptibility testing in 1995. After a period of relative stability in 1981 and 1987-1990, beta-lactamase production increased in H. influenzae. Among middle ear isolates from children under 6 years, beta-lactamase production increased from 8% to 24% in H. influenzae and from 81% to 96% in M. catarrhalis since the survey in 1987-1990. 1.2% of S. pneumoniae were penicillin-resistant and 4.2% intermediately resistant; 5 years earlier among otitis isolates of children only 1.7% intermediate resistance was found. Ampicillin resistance was seen among 1.9% of non-beta-lactamase-producing strains of H. influenzae. Resistance to trimethoprim-sulphamethoxazole occurred in 9.4% of S. pneumoniae, 7.4% of H. influenzae and 0.7% of M. catarrhalis. Frequencies of azithromycin resistance were 3.0% in S. pneumoniae and 1.6% in H. influenzae, and those of tetracycline resistance were 6.7% in S. pneumoniae and 1.2% in H. influenzae. PMID- 9338493 TI - Plasmid-encoded fosfomycin resistance in bacteria isolated from the urinary tract in a multicentre survey. AB - Sixty out of 219 fosfomycin-resistant bacteria selected from more than 7400 urinary pathogens in an epidemiological multicentre survey performed in Italy were screened for plasmid genes fosA and fosB conferring fosfomycin resistance. Only five strains, three enterobacteria and two staphylococci, carried plasmids harbouring, respectively, fosA and fosB genes. Fosfomycin resistance in the other isolates was caused by an alteration of the chromosomally encoded GlpT transport system. One strain, Morganella morganii 279, incorporated alpha-glycerolphosphate and its mechanism of fosfomycin resistance needs to be further investigated. Our study showed that PCR amplification is the most accurate, simple and rapid method for epidemiological studies of plasmid-encoded fosfomycin resistance, and that fosfomycin resistance conferred by plasmid genes (both fosA and fosB) accounts for only a low percentage of the fosfomycin-resistant strains. PMID- 9338494 TI - Correlation between in-vitro susceptibility testing to itraconazole and in-vivo outcome of Aspergillus fumigatus infection. AB - Given the increased choice of therapeutic agents and the rising incidence of serious invasive disease, it is important that reliable in-vitro methods for detecting antifungal drug resistance in Aspergillus spp. are developed. Six clinical isolates of Aspergillus fumigatus, obtained from patients in whom the clinical outcome was known, were selected for study. Each was used to examine a range of parameters affecting agar dilution and broth microdilution susceptibility test results. The in-vitro results were compared with outcome in a neutropenic mouse model of invasive aspergillosis. Groups of animals were treated with itraconazole at 25 mg/kg and 75 mg/kg and survival rates and organ burdens were determined. Itraconazole was efficacious against four isolates (susceptible) but failed for two (resistant) in the animal model of infection. Both the resistant isolates had been obtained from patients receiving itraconazole treatment with good serum concentrations of the drug. Conditions for the agar dilution test which produced results that correlated best with our in-vivo observations included the use of RPMI agar with L-glutamine buffered to pH 7 with MOPS, inoculated with 10(6)-10(7) conidia/mL and incubated for 48-72 h at 28 or 35 degrees C with a no-growth endpoint. Optimal conditions for the broth microdilution method included the use of RPMI medium with L-glutamine and 2% glucose buffered to pH 7 with MOPS, an inoculum of 2 x 10(5) conidia in 200 microL incubated for 48 h at 35 degrees C with a growth (or trace) endpoint. The MICs for the susceptible isolates were 0.12-1.0 mg/L and > or = 16 mg/L for the resistant isolates. With careful selection and standardization of test conditions it is possible to generate reproducible in-vitro susceptibility data for Aspergillus spp. that will predict clinical outcome. PMID- 9338495 TI - Influence of growth rate and nutrient limitation on susceptibility of Burkholderia cepacia to ciprofloxacin and tobramycin. AB - The influence of growth rate and oxygen availability on ciprofloxacin and tobramycin sensitivity and cell surface hydrophobicity in Burkholderia cepacia was assessed for cells grown in a chemostat under iron-limitation. Whereas susceptibility to both antibiotics decreased with increasing growth rate and oxygen repletion yet increased under oxygen depletion, hydrophobicity decreased with increasing growth rate and oxygen repletion but was relatively unaffected under oxygen depletion. These results emphasize the modulating effect of the growth environment on antibiotic susceptibility and the need to perform antimicrobial susceptibility tests under conditions which closely mimic, in vitro, bacterial growth in vivo. PMID- 9338496 TI - Antibacterial efficacy of tobramycin against anaerobic Escherichia coli cultures in the presence of electron acceptors. AB - The antimicrobial activity of tobramycin against anaerobic cultures of Escherichia coli was tested in the presence of various electron carriers. The presence of 2,6-dichlorophenol 4-indophenol (DCIP) significantly enhanced the killing efficacy of tobramycin. Only 0.003% of the initial cell population (i.e. 10(6) cfu/mL) remained viable after exposure for 10 h to the mixture of antibiotic (20 x MIC, i.e. 40 mg/L) and electron acceptor (10(-3) M), as compared with 9% of surviving organisms in the presence of tobramycin alone. Less synergy was obtained with p-benzoquinone and 1,2-naphthoquinone. Fumarate did not affect the efficiency of the antibiotic. The mixture of tobramycin and DCIP was ineffective against agar-entrapped bacteria which, like biofilm organisms, are subject to oxygen limitation. PMID- 9338497 TI - Activity of a new glycopeptide antibiotic (LY333328) against enterococci and other resistant gram-positive organisms. AB - A new semi-synthetic glycopeptide, LY333328, was tested for in-vitro activity against 197 strains of enterococci including strains resistant to other glycopeptides. Activity was also assessed against species with intrinsic resistance to glycopeptides. For strains of enterococci tested the LY333328 MIC90s were < or = 0.5 mg/L. Enterococcus faecalis was less susceptible to LY333328 than other enterococci with a maximum MIC of 2.0 mg/L. Glycopeptide resistant strains of enterococci were not more resistant to LY333328 than glycopeptide-susceptible strains but intrinsically resistant species had MICs between 4 and 8 mg/L. PMID- 9338498 TI - In-vitro activities of ciprofloxacin, levofloxacin, lomefloxacin, ofloxacin, pefloxacin, sparfloxacin and trovafloxacin against gram-positive and gram negative pathogens from respiratory tract infections. AB - Trovafloxacin, sparfloxacin, ciprofloxacin and levofloxacin were equally active against Moraxella catarrhalis, Haemophilus influenzae, Legionella pneumophila, Klebsiella pneumoniae, Enterobacter cloacae and Serratia marcescens. Ciprofloxacin was the most active compound against Pseudomonas aeruginosa (MIC90 = 1 mg/L), followed by trovafloxacin (MIC90 = 4 mg/L). Trovafloxacin was twice as active as sparfloxacin against Streptococcus pyogenes (MIC90 = 0.12 mg/L), Streptococcus pneumoniae (MIC90 = 0.12 mg/L) and Staphylococcus aureus (MIC90 = 0.06 mg/L) (except quinolone-resistant, methicillin-resistant S. aureus, for which the MIC90 was 8 mg/L). Trovafloxacin was the most active compound against Enterococcus faecalis: 80% of strains were susceptible to 0.25 mg/L. There was complete cross-resistance between all fluoroquinolones. PMID- 9338499 TI - The postantibiotic effect of fusidic acid against gram-positive bacteria. AB - The in-vitro postantibiotic effect (PAE) of fusidic acid was tested for six strains of Staphylococcus aureus and four strains of Streptococcus pyogenes. The maximum PAE that could be achieved was more than 3 h, but at concentrations of antibiotic attainable in humans, the PAE was less than 2 h. Additional experiments were performed in albumin, to examine the effect of the strong protein binding of fusidic acid on the MIC and PAE. MICs were increased, but at the same multiple of the MIC, PAEs were similar to those performed in Mueller Hinton broth. PMID- 9338500 TI - In-vitro antimicrobial activity of HSR-903, a new fluoroquinolone, against clinical isolates of Neisseria gonorrhoeae with quinolone resistance-associated alterations in GyrA and ParC. AB - The in-vitro antimicrobial activity of HSR-903, a new fluoroquinolone, was tested against 51 clinical Neisseria gonorrhoeae isolates in comparison with ciprofloxacin, levofloxacin and sparfloxacin. The MICs of HSR-903 for 11 isolates with alterations in both GyrA and ParC, for 19 isolates with alterations only in GyrA and for 21 isolates without alterations in either GyrA or ParC ranged from 0.03 mg/L to 1.0 mg/L (MIC90 = 0.25 mg/L), from 0.03 mg/L to 0.5 mg/L (MIC90 = 0.125 mg/L) and from < or = 0.001 mg/L to 0.008 mg/L (MIC90 = 0.004 mg/L), respectively. Levofloxacin and ciprofloxacin were the least active of the four quinolones tested, particularly against the mutant strains. Sparfloxacin was more active, but HSR-903 exhibited the most potent in-vitro activity against the clinical N. gonorrhoeae isolates, including those harbouring quinolone-resistance associated alterations in GyrA and ParC. PMID- 9338501 TI - Therapeutic effects of omoconazole nitrate on experimental tinea pedis, an intractable dermatophytosis, in guinea-pigs. AB - The therapeutic efficacy of omoconazole nitrate was investigated in an experimental tinea pedis model produced by topical inoculation with Trichophyton mentagrophytes in guinea-pigs, which is pathologically similar to naturally infected tinea pedis in humans. Treatment with omoconazole nitrate cream was started on week 2 postinfection and continued for 3 or 4 weeks. Once-a-day application of 1% omoconazole nitrate to the site of infection exhibited an excellent therapeutic efficacy, and was superior to 1% bifonazole cream in culture result. This result suggests that omoconazole nitrate has a potential usefulness for the treatment of tinea pedis in humans. PMID- 9338502 TI - Concentrations of prophylactic ceftriaxone in abdominal tissues during pancreatic surgery. AB - Ceftriaxone concentrations in abdominal tissues were evaluated after administration as antibiotic prophylaxis for pancreatic surgery. Ten patients were given ceftriaxone (1 g i.v.) 30 min before surgery. Ceftriaxone concentrations in fatty tissues ranged from 2.5 to 6.2 microg/g. Ceftriaxone concentrations were 6.0 +/- 8.6 microg/g in pancreatic tissues, 2.1 +/- 2.5 mg/L in pancreatic fluid, 1179 +/- 1271 mg/L in pancreatic bile, and 18 +/- 16 microg/g in the liver. In fatty tissues, 8-10 patients had tissue levels greater than the MIC90 for Staphylococcus aureus and Escherichia coli and the 10 patients had tissue levels greater than the MIC90 for Klebsiella pneumoniae and Proteus mirabilis. In other tissues, penetration was greater than the MIC90 for potential pathogens in 50-100% of the patients. PMID- 9338503 TI - Teicoplanin in the treatment of enterococcal endocarditis: clinical and microbiological study. AB - Seven cases of enterococcal endocarditis treated with teicoplanin (7-10 mg/kg/day for 28-105 days) alone (one case) or in combination with aminoglycosides (six cases) were reviewed. All patients were cured. Serum bactericidal activity titres after intravenous gentamicin (5 mg/kg every 24 h) and teicoplanin (10 mg/kg every 24 h) were measured on day 7 of treatment in four patients against five enterococcal isolates: mean titres were 1:54 (range 1:16-64) and 1:22 (1:8-32) at 0.5 and 24 h post-infusion, respectively. Time-kill studies showed synergy between teicoplanin and gentamicin against three isolates. We conclude that single daily-dose teicoplanin/gentamicin combined therapy may represent a rational alternative to standard penicillin/gentamicin therapy and a useful regimen for home treatment of selected cases of enterococcal endocarditis. PMID- 9338504 TI - Antimicrobial resistance--relation to human and animal exposure to antibiotics. PMID- 9338506 TI - Activity of meropenem against Enterococcus faecalis. PMID- 9338505 TI - High prevalence of colonization with vancomycin- and pristinamycin-resistant enterococci in healthy humans and pigs in The Netherlands: is the addition of antibiotics to animal feeds to blame? PMID- 9338507 TI - A survey of susceptibility to erythromycin amongst Streptococcus pyogenes isolates in Italy. PMID- 9338508 TI - The effect of reserpine, an inhibitor of multi-drug efflux pumps, on the in-vitro susceptibilities of fluoroquinolone-resistant strains of Streptococcus pneumoniae to norfloxacin. PMID- 9338509 TI - Development of resistance to ciprofloxacin in Acinetobacter baumanii strains isolated during a 20-month outbreak. PMID- 9338510 TI - Renal function after myocardial infarction and cardiac arrest in rats: role of ANP-induced albuminuria? AB - Renal function was measured by clearance technique before and after acute myocardial infarction (MI) induced by left coronary artery ligation in male Sprague-Dawley rats. The animals were anaesthetized with halothane-nitrous oxide, paralysed with pancuronium and artificially ventilated. All parameters were stable throughout the experiment in sham-operated time control animals (n = 8). After MI, rats developed left ventricular dysfunction with increased left ventricular end-diastolic pressure and decreased mean arterial pressure. MI produced antidiuresis and antinatriuresis without changes in glomerular filtration rate (GFR), lithium clearance or renal albumin excretion (n = 8). The antidiuretic and antinatriuretic responses to MI were similar in rats with chronic bilateral renal denervation (n = 5). Three additional rats with chronic bilateral renal denervation had cardiac arrest and were resuscitated with cardiac massage, i.v. lidocaine and intracardiac adrenaline administration. These animals showed a transient increase in urine flow rate, sodium and albumin excretion with maximum 30-60 min after resuscitation, while GFR and lithium clearance were normal. Since cardiac ischaemia and sympathetic stimulation are strong stimuli for the release of atrial natriuretic peptide (ANP), we examined if ANP (0.25, 0.50, and 1.00 microg kg(-1) min(-1), n = 8 per dose) affects urinary albumin excretion. ANP increased dose-dependently the urine/plasma concentration ratio of albumin relative to inulin, which suggests that ANP increases the glomerular permeability for albumin. We conclude that MI causes stimulation of renal tubular sodium and water reabsorption by a mechanism which is independent of intact renal innervation. MI does not produce any change in renal albumin excretion in rats, but transient albuminuria may be observed in rats following cardiac arrest and/or manoeuvres used in cardiac resuscitation. Since ANP produces albuminuria, we speculate that ANP may be an important mediator of albuminuria in states with elevated plasma concentrations of ANP. PMID- 9338511 TI - The natriuretic response to a dopamine DA1 agonist requires endogenous activation of dopamine DA2 receptors. AB - Dopamine produced in the kidney acts as a natriuretic hormone by inhibiting tubular Na+,K+-ATPase activity. Previous in vitro studies have shown that Na+,K+ ATPase activity in the proximal tubule is inhibited by a synergistic action of dopamine 1 (DA1) and dopamine 2 (DA2) receptors. This in vivo study, performed on rats, investigates whether the natriuretic response to DA requires a synergistic action of DA1 and DA2 receptors. The DA1 agonist, fenoldopam, significantly increased urinary sodium excretion, but there was no increase in sodium excretion when a DA1 agonist was given together with a DA2 antagonist. Neither DA1 nor DA2 antagonists had any influence on sodium excretion. The natriuretic response to fenoldopam was also significantly attenuated after the administration of benserazide, which inhibits aromatic acid decarboxylase and thereby suppresses the endogenous production of dopamine. In conclusion, the natriuretic effect of dopamine depends on the activation of both DA1 and DA2 receptors. The DA2 receptor appears to be constitutively activated by endogenous dopamine. PMID- 9338512 TI - Altered basal and adenosine-mediated protein flux from coronary arterioles isolated from exercise-trained pigs. AB - Solute flux per unit surface area and concentration gradient, (J(S)/SdeltaC), was quantified in arterioles isolated from hearts of sedentary (SED) and exercise trained (EX) female Yucatan Miniature Swine. Apparent permeability (P(S)) was assessed from measures of J(S)/SdeltaC for two proteins, alpha-lactalbumin (alpha lact) and porcine serum albumin (PSA), under basal conditions and following 5 min suffusion with 10(-5) M adenosine (ADO). Both proteins were labelled with the fluorescent dye tetramethyl rhodamine isothiocyanate. Basal P(S) to alpha-lact differed with exercise training ((P(S)alpha-lact)SED = 5.2+/-1.8 (median+/-median absolute deviation (MAD), n = 9 pigs) versus (P(S)alpha-lact)EX = 7.4+/-1.1 x 10( 7) cm s(-1), n = 9, P < 0.05). For the larger protein PSA, basal P(S) did not change with training (P(S)PSA)SED = 50+/-1.6, N = 11 vs. (P(S)PSA)EX = 4.1+/-1.2 x 10(-7) cm s(-1), N = 11). Suffusion of the arterioles (33+/-4 microm diameter, n = 18 vessels) from SED hearts (n = 14) with 10(-5) M ADO decreased P(S)alpha lact 15+/-8% relative to control and was without effect on P(S)PSA. By contrast, in arterioles (39+/-4 microm diameter, n = 22 vessels) from EX hearts (n = 14), ADO increased P(S)alpha-lact and P(S)PSA by 32 and 65% respectively, indicating that receptor-mediated changes in permeability were also sensitive to exercise training. These data demonstrate that, for coronary arterioles, permeability to macromolecules adapts to exercise training. The adaptive mechanisms may involve more than one structural component of the vessel wall as the changes in permeability were size-dependent. PMID- 9338513 TI - Adaptation of myoglobin in compensatory hypertrophied rat muscle. AB - This study investigated the time course change of myoglobin concentration ([Mb]) in skeletal muscle during muscle hypertrophy in rats. Seven groups of Wistar rats (n = 7 per group) were examined. Compensatory hypertrophy of the plantaris muscle was induced by the bilateral ablation of the medial and lateral heads of the gastrocnemius muscle, and the effect of compensatory hypertrophy on the [Mb] of the plantaris muscle was examined at 7, 21 and 42 days post-ablation. The [Mb] expressed as mg g(-1) wet muscle weight tended to decline at 7 days post-ablation (-12.1%) and returned to the control level by 42 days post-ablation, as did the muscle protein concentration. However, the [Mb] expressed as mg g(-1) protein weight was not significantly different between the ablation and control groups throughout the experimental period. The estimated value of absolute Mb content in the whole plantaris muscle was significantly increased by 28.4% (P < 0.01) at 42 days post-ablation compared with the age-matched controls. Therefore, the longer period of compensatory activity allowed the Mb to increase in its absolute content, although the [Mb] did not exceed the level of control muscle. The capacity of facilitated oxygen diffusion and oxygen storage by Mb in the muscle cells will not change even when muscle enlargement takes place. PMID- 9338514 TI - Duration and mechanisms of the increased natural cytotoxicity seen after chronic voluntary exercise in rats. AB - We have recently shown that in vivo natural cytotoxicity is enhanced after chronic exercise in spontaneously hypertensive rats (SHRs). In the present report, we have studied the duration of this augmentation and some possible mechanisms involved. Exercise consisted of voluntary running for 4-5 weeks, with the running distance ranging from 2.7-15.6 km day(-1) during the last week of running. In vivo cytotoxicity was measured as clearance of injected 51Cr-labelled YAC-1 lymphoma cells from the lungs. The in vivo natural cytotoxicity was increased in running SHRs, and also in SHRs that had their running wheel locked for 24 and 48 h prior to the experiment, and was still present after 96 h. The enhancement of in vivo cytotoxicity after 5 weeks of exercise was abolished after an acute injection of the beta-adrenergic receptor antagonist timolol (0.5 mg kg( 1) i.v.), indicating that catecholamines are involved in this augmentation. Interestingly, 24 h after the last exercise bout, the increased natural cytotoxicity could be blocked by timolol. The opioid receptor antagonist naloxone given subcutaneously for 7 days by osmotic pumps (6 mg kg(-1) h(-1)) could not reverse the increased in vivo cytotoxicity seen in the running SHRs, suggesting that opioid receptor mechanisms are not involved, or at least not the naloxone sensitive mu-receptor. Natural immunity was not influenced by the histamine H2 receptor antagonist ranitidine, either in controls or in runners, indicating that the natural killer cell-regulatory effect of histamine is not present in SHRs and does not seem to be involved in the exercise-induced changes in natural immune function. We conclude that the augmentation of in vivo natural cytotoxicity after voluntary chronic exercise in rats is long-lasting and that the augmentation is partly mediated by beta-adrenergic receptors. PMID- 9338515 TI - The relationships between EMG and muscle morphology throughout sustained static knee extension at two submaximal force levels. AB - This study investigated the relationship between muscle morphology and surface electromyographic parameters (mean frequency, f(mean); and signal amplitude, RMS) during sustained static knee extension to exhaustion at 25% maximal voluntary contraction (MVC) and at 70% MVC. Twenty clinically healthy subjects participated. EMGs were registered from the quadriceps. Muscle forces during knee extension at a 90 degree joint angle were maintained at the respective levels throughout the measurement periods. A biopsy was obtained of the vastus lateralis muscle. The total time to exhaustion was normalized for each subject. By means of regression analysis, the intercept (i) (i.e. the unfatigued state) and the regression coefficient (k) were determined for each EMG parameter. The endurance time increased with decreasing force level. A significantly higher perception of fatigue was found at 25% MVC than at 70% MVC. From principal component analyses it was concluded that RMS-k at 25% MVC mainly correlated with the type 2 muscle fibre proportions (%), and at 70% MVC mainly with the areas of type 2 muscle fibre. At 25% MVC, f(mean)-k correlated with the areas of type 2A, 2B and 2C fibres, and at 70% MVC negatively with the proportion of type 2B and to some extent with areas of type 2A, 2B and 2C fibres. f(mean)-i at 25% MVC correlated with fibre type proportions (%); f(mean)-i at 70% MVC correlated with the areas of type 2A, 2B and 2C. The present study indicates relationships between surface EMG and muscle morphology, which is contrary to presented models of the EMG. PMID- 9338516 TI - Differential responses in intramuscular pressure and EMG fatigue indicators during low- vs. high-level isometric contractions to fatigue. AB - This study investigated changes in intramuscular pressure (IMP) and surface electromyographic (EMG) parameters (mean frequency of the power spectrum, f(mean); and signal amplitude denoted as root mean square, RMS) during contractions to fatigue at 25 and 70% of maximal voluntary contraction (MVC). Parameters were recorded simultaneously from the vastus lateralis muscle during knee extension. A significant decrease in f(mean) occurred with time at both contraction levels; however, the rate of decline (slope) was greater at 70% MVC. RMS increased throughout the contractions at both levels, with the relative increase being significantly greater for 25% MVC. IMP increased during 25% MVC but did not change during the 70% MVC contraction. IMP at rest was significantly higher post-contractions than it was pre-contractions at 25% MVC (21.1 vs. 8.0 mmHg, P < 0.01) and 70% MVC (13.7 vs. 8.6 mmHg, P < 0.01). Consequently, post contraction IMP was higher at 25% MVC than at 70% MVC (P < 0.01). IMP changes throughout the fatiguing contractions correlated negatively with f(mean) and positively with RMS at both MVC levels; however, these correlations were better at 25% MVC. The extent of intramuscular water accumulation is discussed as a major cause of the difference in IMP changes between 25% and 70% MVC. Significant differences in the rate of change for all parameters between high vs. low contraction levels may suggest a common mechanism governing changes in IMP and EMG fatigue indicators. PMID- 9338517 TI - Acid-induced increase in duodenal mucosal permeability is augmented by nitric oxide inhibition and vasopressin. AB - The aim of the study was to determine if and by what mechanism(s) nitric oxide inhibition modulates the susceptibility of the duodenum to hydrochloric acid induced disturbances of mucosal integrity. A second aim was to investigate whether basal permeability is a determinant of epithelial acid barrier function. Using an in situ duodenal perfusion model, mucosal permeability, alkaline secretion and morphology were investigated in anaesthetized rats. Luminal perfusion with 50 mM hydrochloric acid increased duodenal mucosal permeability in the control animals. In animals receiving the nitric oxide synthase inhibitor N nitro-L-arginine methyl ester (L-NAME 3 mg kg(-1) and 1 mg kg(-1) h(-1)) and in those receiving vasopressin (1 IU kg(-1) h(-1)), however, the mean increase in permeability in response to acid was markedly higher. In rats treated with either hexamethonium (20 mg kg(-1)) or atropine (0.5 mg kg(-1)) L-NAME failed to augment the acid-induced increase in permeability. Perfusion with hypotonic saline (25 mM) increased basal permeability but did not influence the response to acid. Exposure of the duodenum to hydrochloric acid caused very subtle changes of duodenal morphology. It is concluded that both inhibition of endogenous nitric oxide synthesis and vasopressin treatment augment the acid-induced increase in mucosal permeability. The mechanisms involved may be related to changes of Starling forces in the microcirculatory bed. Endogenous nitric oxide may protect the duodenal mucosa by regulating vascular permeability and interstitial fluid pressure. PMID- 9338519 TI - Effects of calcium channel blockade on intestinal fluid secretion: sites of action. AB - Most intestinal secretagogues, including cholera toxin, evoke fluid secretion in part by activating the enteric nervous system (ENS). The enterotoxins that, due to size, cannot pass the intestinal epithelial lining have been proposed to activate the ENS via the release of amines/peptides from the intestinal endocrine cells. It has been shown that calcium channel blockers of the L-type attenuate intestinal fluid secretion. This study was performed on rat jejunal segments to elucidate where calcium channel antagonists interact with the secretory nervous reflex(es) of the ENS. In vivo, net fluid transport, transmural potential difference (PD) and luminal release of serotonin from the enterochromaffin cells were monitored before and after exposing the intestinal mucosa to cholera toxin (20 microg/mL) or the calcium ionophore A23187 (0.5 mM). In vitro, the effects of transmural electrical field stimulation (EFS) on short circuit current (SCC) was investigated using the Ussing chamber method. Cholera toxin and A23187 evoked a net fluid secretion, an increased PD and an augmented luminal release of 5-HT. These effects were markedly attenuated by giving the calcium channel blocker nifedipine i.v. (5.75 micromol kg(-1) body wt). On the other hand, nifedipine (0.02 mM) had no significant effect on the increased SCC caused by EFS in vitro. The results obtained in the in vivo experiments suggest that the nifedipine markedly attenuates the initial event in cholera toxin- and A23187-induced secretion, the release of amines and probably also of peptides from the intestinal endocrine cells. The in vitro experiments seem to exclude an effect of the calcium channel blockade on the efferent part of the secretory nervous reflex. PMID- 9338518 TI - Calcium channels and intestinal fluid secretion: an experimental study in vivo in rats. AB - Several mechanisms involved in nervous secretory reflex(es) of the enteric nervous system may be dependent on the flux of calcium across the plasma membrane, which may be controlled by voltage-gated calcium channels. In this study, we investigated the importance of plasma membrane calcium channels for intestinal fluid secretion. Two types of studies were performed, in which intestinal net fluid transport in anaesthetized rats was followed with a gravimetric method. First, the effects on intestinal fluid transport of placing A23187, a calcium ionophore, in the intestinal lumen was studied. A23187 induced in a dose-dependent manner a net fluid secretion, which was abolished by giving hexamethonium (10 mg kg(-1) body wt) i.v. or placing lidocaine (1% solution) on the intestinal serosa. Nifedipine (5.75 micromol kg(-1) body wt i.v.) also abolished the fluid secretion caused by the ionophore. In the second study, the effects of various calcium channel blockers (gadolinium chloride, nifedipine, verapamil) were tested on the cholera toxin-induced secretion. It was attenuated by luminal application of gadolinium chloride (1-10 mM) or nifedipine (10-200 microM). Intravenously administered nifedipine (2.5-5.75 micromol kg(-1) body wt) abolished cholera toxin-evoked secretion dose-dependently, whereas verapamil (0.05-1 micromol kg(-1) body wt) was without consistent effect. It is concluded that the fluid secretion evoked by placing A23187 in the intestinal lumen in vivo was induced via an activation of the enteric nervous system. Cholera secretion was attenuated or abolished by calcium channel blockers of the L- or N-type. PMID- 9338520 TI - Effects of topical platelet activating factor on the guinea-pig tracheobronchial mucosa in vivo. AB - Platelet activating factor (PAF) has been reported to produce a variety of airway effects including epithelial damage and increased airway-lung absorption of hydrophilic tracers. The present study examines effects of PAF on the guinea-pig tracheobronchial mucosa in vivo. Vehicle with and without PAF (4.0 and 8.0 nmol) was superfused onto the tracheobronchial mucosa. The levels of 125I-albumin, previously given intravenously, were determined in tracheobronchial lavage fluids as an index of mucosal exudation of plasma. The mucosa was also examined by scanning electron microscopy. In separate animals, 99mTc-DTPA (a low molecular weight, 492 Da, hydrophilic tracer) was superfused onto the mucosal surface through an oro-tracheal catheter, together with vehicle or PAF (8.0 nmol). A gamma camera determined the disappearance rate of 99mTc-DTPA from the airways as an index of mucosal absorption. PAF produced dose-dependent mucosal exudation of plasma up to 20-fold greater than control (P < 0.001). However, PAF did not damage the epithelium and the absorption ability of the airway mucosa was unaffected. The results, in contrast to previous reports, suggest that PAF may not readily damage the airway mucosa even at large exudative doses of the agent. The present finding support the view that the plasticity of the epithelial junctions allows the creation of valve-like paracellular pathways for unidirectional clearance of extravasated plasma into the airway lumen. We suggest that endogenous PAF may participate in first line respiratory defence reactions by causing lumenal entry of bulk plasma without harming the epithelium. PMID- 9338521 TI - The cGMP pathway is not responsible for the blunted hypoxic vasoconstriction in rat lungs after altitude exposure. AB - To examine the contribution of the cyclic guanosine monophosphate (cGMP) pathway in changes in pulmonary vasoconstriction during the initial days of altitude exposure, we tested the effects of LY83583 (an inhibitor of guanylate cyclase activation) and those of N(G)-monomethyl-L-arginine (an inhibitor of nitric oxide synthesis) on airway hypoxia- (3% O2) and angiotensin II- (AII, 0.2 microg) induced vasoconstrictions in lungs from the rats exposed to either moderate altitude (MA, 570 torr) or high altitude (HA, 430 torr) At 2 days' exposure, hypoxic response was significantly blunted compared with the response in low altitude (LA, 710 torr) lungs in an altitude-dependent manner. At 7 days' exposure, the response was recovered fully in MA lungs but partially in HA lungs. AII response was not significantly blunted at 2 days' exposure, but was significantly augmented in an altitude-dependent manner at 7 days' exposure. LY83583 (10 micromol L(-1)) potentiated both responses in LA lungs but did not significantly potentiate either response in any altitude-exposed lungs. N(G) monomethyl-L-arginine (10 micromol L(-1)) potentiated both responses in LA lungs but did not significantly potentiate either response in HA lungs at 2 days' and 7 days' exposure. Thus the cGMP pathway is not responsible for either the change in hypoxic vasoconstriction or the change in AII vasoconstriction in rat lungs during the initial 7 days of altitude exposure. PMID- 9338522 TI - Inhaled carbon dioxide inhibits lower airway nitric oxide formation in the guinea pig. AB - The present study addressed the effect of inhaled carbon dioxide on lower airway nitric oxide formation in normoxic anaesthetized guinea pigs. Ventilation with carbon dioxide (1.5, 3, 6, 9 and 12%) induced a concentration-dependent decrease in the basal concentration of nitric oxide in exhaled tracheal air. A maximal reduction in exhaled nitric oxide of approximately 25% was induced by 12% carbon dioxide in inhaled air. Ventilation with positive end-expiratory pressure (7 cmH2O) increased the concentration of exhaled nitric oxide. Inhalation of carbon dioxide had a larger, concentration-dependent, inhibitory effect (maximally 60%) on the lower airway nitric oxide formation induced by ventilation with positive end-expiratory pressure, as compared with the effect on the basal concentration of nitric oxide. The results show that inhaled carbon dioxide suppresses lower airway nitric oxide excretion in the guinea pig. Endogenous carbon dioxide might exert effects through regulation of endogenous nitric oxide formation, for example in the regulation of airway tone or in ventilation-perfusion matching. PMID- 9338523 TI - Quantification of alpha2A and alpha2C adrenoceptors in the rat striatum and in different regions of the spinal cord. AB - The alpha2C-adrenoceptor preferring radioligand [3H]-MK912 was used for labelling alpha2A- and alpha2C-adrenoceptors in the rat striatum, in the cervical, thoracic and lumbar parts of the spinal cord, and in the dorsal and ventral halves of the spinal cord. In addition, guanfacine was used as a tool to delineate the alpha2A- and alpha2C-adrenoceptors. In the striatum the sites were 72% alpha2A- and 28% alpha2C-adrenoceptors, while in all regions of the spinal cord the proportions of the sites were about 96% alpha2A- and 4% alpha2C-adrenoceptors. A multi-curve experimental design and computer analysis was used in order to enable the accurate quantification of the alpha2A- and alpha2C-adrenoceptors in the striatum and spinal cord. PMID- 9338524 TI - Glucagon-like peptide-1 reduces hepatic glucose production indirectly through insulin and glucagon in humans. AB - The effect of glucagon-like peptide-1 (GLP-1) on hepatic glucose production and peripheral glucose utilization was investigated with or without infusion of somatostatin to inhibit insulin and glucagon secretion in 13 healthy, non diabetic women aged 59 years. After 120 min 3-(3)H-glucose infusion, GLP-1 was added (4.5 pmol kg(-1) bolus + 1.5 pmol kg(-1) min(-1)). Without somatostatin (n = 6), GLP-1 decreased plasma glucose (from 4.8 +/- 0.2 to 4.2 +/- 0.3 mmol L(-1), P = 0.007). Insulin levels were increased (48 +/- 3 vs. 243 +/- 67 pmol L(-1), P = 0.032), as was the insulin to glucagon ratio (P = 0.044). The rate of glucose appearance (Ra) was decreased (P = 0.003) and the metabolic clearance rate of glucose (MCR) was increased during the GLP-1 infusion (P = 0.024 vs. saline). Also, the rate of glucose disappearance (Rd) was reduced during the GLP-1 infusion (P = 0.004). Since Ra was reduced more than Rd, the net glucose flow was negative, which reduced plasma glucose. Somatostatin infusion (500 microg h(-1), n = 7) abolished the effects of GLP-1 on plasma glucose, serum insulin, insulin to glucagon ratio, Ra, Rd, MCR and net glucose flow. The results suggest that GLP 1 reduces plasma glucose levels mainly by reducing hepatic glucose production and increasing the metabolic clearance rate of glucose through indirectly increasing the insulin to glucagon ratio in healthy subjects. PMID- 9338525 TI - What health services researchers need to know about the Health Insurance Portability and Accountability Act. PMID- 9338526 TI - Counseling patients about prescribed medication: 12-year trends. AB - OBJECTIVES: The authors determined patients' report of prescription drug counseling activities after withdrawal of the pilot program to require patient package inserts in 1980 and implementation of Omnibus Budget Reconciliation Act of 1990 counseling requirements in 1993. METHODS: Four cross-sectional national telephone surveys were conducted in the fall of 1982, 1984, 1992, and 1994. Telephone households were chosen by random-digit dialing. Subjects had obtained a new prescription for themselves or for a family member at a retail pharmacy during the previous 4 weeks. Verbal counseling rates at physician offices and pharmacies for five information categories and the distribution of written information at those locations were determined. RESULTS: Spontaneous verbal counseling at the physician's office has increased slightly, with the largest increases focused on the delivery of side effect and precautionary information. Slightly larger increases in pharmacy-delivered information regarding directions for use and precautions have occurred. Patient questioning has remained at single digit levels at both sites. The percentage of patients receiving any written information has increased from 5% to 15% at the physician's office and from 16% to 59% at the pharmacy. CONCLUSIONS: The data indicate small increases in verbal counseling but larger increases in the delivery of written information provided at the pharmacy. In light of Healthy People: 2000 goals for patient counseling and legislation encouraging private-sector initiatives, these data should help to refocus attention on the continuing need for effective patient education interventions. PMID- 9338527 TI - Factors affecting primary care physician participation in Medicare. AB - OBJECTIVES: This study investigates the levels of participation and the relative association of economic and noneconomic factors on primary care physician participation in the Medicare program. METHODS: Demographic information, participation in Medicare, and attitudes toward both the Medicare program and Medicare patients were collected in a written survey mailed to half the primary care physicians in Iowa. Ordinary least squares and logistic regression analyses were conducted to determine factors associated with the percentage of Medicare patients in a practice and the acceptance of all new Medicare patients, respectively. RESULTS: Two thirds of physicians were accepting all new Medicare patients, whereas 16% were accepting no new Medicare patients. Factors associated with having a higher percentage of Medicare patients in a practice were as follows: (1) a larger proportion of Medicare recipients in the county, (2) practice as a general internal medicine physician, (3) more years in practice at the current location, (4) greater enjoyment treating elderly patients, (5) less concern about having too many Medicare patients, and (6) a stronger belief that the Medicare program respects their professional judgment. Physicians less concerned about having too many Medicare patients in their practice and physicians in counties with a higher percentage of Medicare patients were significantly more likely to accept all new Medicare patients. CONCLUSIONS: These results suggest that as Medicare reforms are discussed, careful consideration of the impact of these reforms on noneconomic issues is important to ensure adequate physician participation and access for elderly patients through the Medicare program. PMID- 9338528 TI - Using neural networks to identify patients unlikely to achieve a reduction in bodily pain after total hip replacement surgery. AB - OBJECTIVES: Fourteen patient-provided variables were chosen as potential predictors for improvement after total hip replacement surgery. These variables included patient demographic information, as well as preoperative physical function. METHODS: A neural network was trained to predict the relative success of total hip replacement surgery using this presurgical patient survey information. The outcome measure was improvement in the Medical Outcomes Study 36 Short Form Health Survey pain score between the preoperative assessment and the 1 year postoperative assessment. For the study sample, 221 patients were selected who had complete information for the composite outcome variable. A backpropagation feedforward neural network was trained to predict the output variable using the jackknife method. RESULTS: Performance of the neural network was assessed by calculating the area under the receiver operating characteristic curve for the network's ability to predict whether the pain score was improved after total hip replacement surgery. The observed area under the receiver operating characteristic curve was 0.79. For comparison, a linear regression model built using the same data had a receiver operating characteristic area of 0.74 (P = 0.23). CONCLUSIONS: This research therefore showed the ability of neural networks to predict the success of total hip replacement more accurately. Our results further indicate that it may be possible to predict which patients are at greatest risk of a poor outcome. PMID- 9338529 TI - The risk of hospitalization for congestive heart failure among older adults. AB - OBJECTIVES: The purpose of the study was to estimate the 8-year rate of hospitalization for congestive heart failure (CHF), to report the resources consumed, and to evaluate previously reported risk factors in a nationally representative sample of 7,286 older white and black adults. METHODS: Secondary analysis of baseline interview data was linked to Medicare hospitalization and death records for 1984 to 1991. Hospitalization for CHF was defined as having one or more episodes with an International Classification of Diseases (ninth revision, clinical modification) discharge code of 428. Combined and separate analyses of first-listed and second-through fifth-listed CHF discharge diagnoses were conducted. Multivariable proportional hazards models were used to evaluate the risks in pooled analyses of all white and black men and women and in separate stratified analyses of white men and white women. RESULTS: Over the 8-year period, 1,102 or 15.1% of the 7,286 older white and black adults were hospitalized for CHF (7.1% with first-listed and 8.1% with second- through fifth listed diagnoses). The 1- and 5-year combined postdischarge mortality rates were 34.7% and 69.0%, respectively. In descending order, the major risk factors for being hospitalized for CHF in the combined, pooled analysis were age, being a white man, having lower body functional limitations, and having self-reported medical histories of coronary heart disease, heart attack, diabetes, and angina. The increased risk associated with age was not linear, and it diminished significantly over the course of life. Some significant differences were observed in the risk factors for hospitalization for first-listed versus second- through fifth-listed CHF and in the risk factors for white women versus white men. CONCLUSIONS: Hospitalization for CHF among older adults is a common, costly event with a poor prognosis. The differential risk for white men remains unexplained and warrants further study. PMID- 9338530 TI - The association between the quality of inpatient care and early readmission: a meta-analysis of the evidence. AB - OBJECTIVES: To help resolve the current controversy over the validity of early readmission as an indicator of the quality of care, the authors critically reviewed the literature using meta-analysis to derive summary estimates of effect and evaluate inter-study heterogeneity. METHODS: The authors selected reports meeting five criteria: (1) presentation of new data on medical-surgical hospitalization of adults; (2) measurement of outcome as a person-specific readmission; (3) readmission within < or = 31 days; (4) examination of some aspect of the process of inpatient care; (5) inclusion of a comparison group. One meta-analysis examined 13 comparisons of readmission rates after substandard versus normative care, another examined 9 comparisons of readmission rates after normative versus exceptional care, and the third examined all 22 comparisons together. Two authors applied inclusion criteria and extracted data on methods and findings. Two others classified studies on 11 methodological variables for the heterogeneity evaluation. RESULTS: The summary odds ratio for readmission after substandard care was 1.24 (0.99-1.57) relative to normative care; for readmission after normative care the summary odds ratio was 1.45 (0.90-2.33) relative to exceptional care. The individual odds ratios varied significantly (chi2, 21 df = 50.34, P = 0.0003). Most of the variance in study odds ratios could be explained by whether the study focused on the quality of patient care or the qualifications of patient care providers. The summary odds ratio for the 16 homogeneous comparisons focusing on the quality of patient care was 1.55 (1.25 1.92). CONCLUSIONS: Early readmission is significantly associated with the process of inpatient care. The risk of early readmission is increased by 55% when care is of relatively low quality, that is, substandard or normative instead of normative or exceptional. PMID- 9338531 TI - The effects of managed care on administrative burden in outpatient substance abuse treatment facilities. PMID- 9338532 TI - Personal characteristics and self-assessed health. PMID- 9338534 TI - The future of the subspecialty of allergy and immunology. PMID- 9338533 TI - The protooncogene c-kit and c-kit ligand in human disease. PMID- 9338535 TI - Relationship between food-specific IgE concentrations and the risk of positive food challenges in children and adolescents. AB - BACKGROUND: The double-blind, placebo-controlled food challenge (DBPCFC) is the "gold standard" for diagnosis of food hypersensitivity. Skin prick tests and RASTs are sensitive indicators of food-specific IgE antibodies but poor predictors of clinical reactivity. Previous studies suggested that high concentrations of food-specific IgE antibody were predictive of food-induced clinical symptoms. Because the CAP System FEIA (Pharmacia Diagnostics, Uppsala, Sweden) provides a quantitative assessment of allergen-specific IgE antibody, this study was undertaken to determine the potential utility of the CAP System FEIA in diagnosis of IgE-mediated food hypersensitivity. METHODS: Sera from 196 patients with food allergy were analyzed for specific IgE antibodies to egg, milk, peanut, soy, wheat, and fish by CAP System FEIA. Sera were randomly selected from 300 stored samples of children and adolescents who had been evaluated by history, skin prick tests, and DBPCFCs. The study population was highly atopic; all patients had atopic dermatitis, and approximately 50% had asthma and allergic rhinitis at the time of initial evaluation. The performance characteristics of the CAP System FEIA were compared with those of skin prick tests and the outcome of DBPCFCs or "convincing" histories of anaphylactic reactions. RESULTS: The prevalence of specific food allergies in the study population varied from 22% for wheat to 73% for egg. Allergy to egg, milk, peanut, and soy accounted for 87% of confirmed reactions. The performance characteristics of skin prick tests and CAP System FEIA (egg, milk, peanut, fish) were comparable, with excellent sensitivity and negative predictive accuracy but poor specificity and positive predictive accuracy. The performance characteristics of the CAP System FEIA for soy and wheat were poor. For egg, milk, peanut, and fish allergy, diagnostic levels of IgE, which could predict clinical reactivity in this population with greater than 95% certainty, were identified: egg, 6 kilounits of allergen-specific IgE per liter (kU[A]/L); milk, 32 kU(A)/L; peanut, 15 kU(A)/L; and fish, 20 kU(A)/L. CONCLUSIONS: When compared with the outcome of DBPCFCs, results of CAP System FEIA are generally comparable to those of skin prick tests in predicting symptomatic food hypersensitivity. Furthermore, by measuring the concentrations of food-specific IgE antibodies with the CAP System FEIA, it is possible to identify a subset of patients who are highly likely (>95%) to experience clinical reactions to egg, milk, peanut, or fish. This could eliminate the need to perform DBPCFCs in a significant number of patients suspected of having IgE-mediated food allergy. PMID- 9338536 TI - Effectiveness of prophylactic inhaled steroids in childhood asthma: a systemic review of the literature. AB - BACKGROUND: There has been no systematic appraisal of the evidence regarding the effectiveness of prophylactic inhaled steroids in childhood asthma. OBJECTIVE: We sought to evaluate the effectiveness of prophylactic inhaled steroids in childhood asthma. METHODS: A MEDLINE search from January 1966 through December 1996 was used to identify pertinent English-language publications. All randomized, double-blind, placebo-controlled trials of prophylactic inhaled steroid therapy for childhood asthma that included data on clinical outcomes (symptom scores and concomitant drug use) or laboratory outcomes (peak expiratory flow rate) were included. RESULTS: In total, 24 of 93 studies retrieved met the inclusion criteria. The overall weighted relative improvement in mean total symptom score (inhaled steroid vs placebo) was 50% (95% confidence interval [CI]: 49%, 51%), the overall weighted relative decrease in mean concomitant beta2 agonist use (inhaled steroid vs placebo) was 37% (95% CI: 36%, 38%), and the overall weighted relative decrease in mean concomitant oral steroid use (inhaled steroid vs placebo) was 68% (95% CI: 66%, 70%). The overall weighted absolute improvement in mean peak expiratory flow rate (inhaled steroid vs placebo) was 38 L/min (95% CI: 34.3 L/min, 41.7 L/min). CONCLUSIONS: Prophylactic inhaled steroids are effective, compared with placebo, in improving both clinical and laboratory outcomes in childhood asthma. PMID- 9338537 TI - Efficacy of antihistamine pretreatment in the prevention of adverse reactions to Hymenoptera immunotherapy: a prospective, randomized, placebo-controlled trial. AB - BACKGROUND: Some clinical studies suggest that a combination of an H1- and H2 antagonist may be effective in the prophylaxis of allergic reactions. OBJECTIVE: The efficacy of pretreatment with an H1/H2-antagonist combination, H1-antagonist alone, or placebo in the prophylaxis of local and systemic adverse reactions to specific immunotherapy with Hymenoptera venom was compared. METHODS: In a prospective, randomized, double-blind, placebo-controlled study, 121 patients with Hymenoptera venom allergy were treated with rush immunotherapy and pretreatment with one of the following: 120 mg of terfenadine plus 300 mg of ranitidine, 120 mg of terfenadine alone, or placebo. The incidence of unwanted systemic adverse and local reactions was recorded for up to 50 weeks. RESULTS: In seven patients (6%), six in the placebo group and one in the terfenadine group, systemic side effects required cessation of therapy (p = 0.005). Subjective symptoms occurred in four patients (10%) in the terfenadine plus ranitidine group and in three patients (7%) in the terfenadine group. Regarding local reactions, significantly fewer patients treated with a combination of terfenadine and ranitidine and with terfenadine alone as compared with placebo had severe local symptoms of erythema (29%, 29%, and 49%), edema (24%, 18%, and 41%), and pruritus (13%, 11%, and 31%) at week 1 (p < 0.05). This therapeutic benefit was limited to the first 4 weeks of treatment. Treatment with a combination of terfenadine and ranitidine was not superior to treatment with terfenadine alone. CONCLUSIONS: Pretreatment with H1-antihistamines with or without H2-antihistamines significantly reduced local and systemic adverse reactions to immunotherapy with Hymenoptera venom and may therefore be helpful in the management of immunotherapy. PMID- 9338538 TI - A simple washing procedure with eucalyptus oil for controlling house dust mites and their allergens in clothing and bedding. PMID- 9338539 TI - Fluticasone propionate powder administered through Diskhaler versus triamcinolone acetonide aerosol administered through metered-dose inhaler in patients with persistent asthma. AB - BACKGROUND: Attempts to delineate efficacy and safety differences among inhaled corticosteroids have been difficult because of the lack of well-controlled, comparative studies reported in the medical literature. METHODS: A randomized, double-blind, double-dummy study was conducted in 24 outpatient centers. A total of 291 male and female patients at least 12 years of age with asthma (FEV1 between 50% and 80% of predicted value), who had previously received maintenance therapy with beclomethasone dipropionate or triamcinolone acetonide, were switched to treatment with fluticasone propionate powder (250 microg twice daily), triamcinolone acetonide aerosol (200 microg four times daily), or placebo for 24 weeks. RESULTS: Mean increase in FEV1 from baseline to end point was significantly (p = 0.009) greater in patients switched to treatment with fluticasone compared with patients switched to treatment with triamcinolone (0.27 L and 0.07 L, respectively). At end point, mean increase in morning peak expiratory flow from baseline was 21 L/min with fluticasone compared with mean decreases of 6 L/min and 28 L/min with triamcinolone and placebo, respectively (p < 0.001 vs triamcinolone and placebo). Supplemental rescue albuterol use decreased by 30% from baseline with fluticasone (p < 0.05 vs triamcinolone and placebo) compared with triamcinolone (6%) or placebo (increased by 50%). The percentage of patients withdrawn from the study because they met predefined lack of-efficacy criteria was higher with placebo (60%) and triamcinolone (27%) than with fluticasone (17%). Incidence of adverse events and low morning plasma cortisol concentrations were similar across treatment groups except for oral candidiasis (p = 0.035, fluticasone vs placebo). CONCLUSION: Fluticasone propionate powder twice daily (500 microg/day) was superior in efficacy to triamcinolone acetonide aerosol four times daily (800 microg/day) in patients with persistent asthma. PMID- 9338540 TI - Evidence of Hop Japanese pollinosis in Korea: IgE sensitization and identification of allergenic components. AB - BACKGROUND: Hop Japanese (Hop J) pollens are abundant in the air of Korea during the autumn season. Their significance as a source of allergic sensitization is still unclear. OBJECTIVES: We sought to detect the sensitization rate to Hop J pollen as an inhalant allergen and to identify its allergenic components. METHOD: We carried out skin prick tests with Hop J pollen extract in 1287 patients with respiratory allergy who visited our hospital over the course of 1 year. The serum specific IgE antibody to Hop J pollen antigen was detected by ELISA, and its binding specificity was confirmed by the ELISA inhibition test. To confirm the respiratory sensitization, bronchoprovocation tests were performed in 16 asthmatic patients sensitive to this pollen. To characterize allergenic components, Hop J pollen extract was analyzed by means of sodium dodecylsulfate polyacrylamide gel electrophoresis followed by IgE immunoblotting. RESULTS: A positive result on the skin prick test (> or = 2+ of the antigen to histamine ratio) was noted in 79 (6.1%) patients. The serum-specific IgE antibody was detected in 18 (41.9%) patients among 43 positive reactors tested. The ELISA inhibition test with the addition of Hop J pollen extract showed a dose-dependent response. Minimal inhibitions were noted with addition of ragweed and mugwort pollen extracts. Nine asthmatic patients showed significant bronchoconstriction after inhalation of the Hop J pollen extract (five early and four dual asthmatic responders), and all of them had high specific IgE binding. Immunoblot analysis revealed 12 IgE-binding components ranging from 13 to 89 kd. Three bands (13 kd, 74 kd, and 80 kd) were bound to the IgE among the sera tested from more than 50% of the patients. CONCLUSION: We believe that the Hop J pollen should be considered as a relevant allergen during the autumn season and thus included in skin test batteries in Korea. Some patients diagnosed as having "intrinsic" asthma or rhinitis might be sensitized to this pollen and other previously unknown allergens. PMID- 9338541 TI - Proteolytic detergent enzymes enhance the allergic antibody responses of guinea pigs to nonproteolytic detergent enzymes in a mixture: implications for occupational exposure. AB - A guinea pig intratracheal test was developed to assess the respiratory allergenicity of enzymes used in the detergent industry. Information gained from this test was used in a process for setting operational exposure guidelines to protect worker health. Mixtures of enzyme proteins were given to guinea pigs once per week for 10 weeks to determine whether there were interactions among enzymes that affected the induction of antibody responses to each enzyme in the mixture. Passive cutaneous anaphylaxis antibody titers against each enzyme were measured in sera. Mixtures of two or three enzymes always consisted of a protease (Alcalase, Savinase; Novo Industri A/S) with an alpha-amylase (Termamyl; Novo Industri A/S), a lipase (Lipolase; Novo Industri A/S), or both. Control animals were exposed to single enzymes. The antibody titers to Termamyl and Lipolase were significantly greater in animals dosed with the protease-containing mixtures as compared with control animals dosed with a single enzyme. Antibody titers to the protease were unchanged in the presence of additional enzymes in the mixture. Complete inactivation of protease activity abrogated the enhanced antibody response to Lipolase. Inhalation exposure of guinea pigs to a mixture of Alcalase and Lipolase also resulted in higher antibody titers to Lipolase as compared with animals exposed by inhalation to Lipolase alone, showing that the enhanced response was not due to intratracheal delivery of antigen to the respiratory tract. These results show that proteolytic enzymes in a mixture enhance antibody responses to other enzymes. This should be considered when defining exposure guidelines for protease-containing enzyme mixtures. PMID- 9338542 TI - Reliability of the Medtrac MDI Chronolog. AB - BACKGROUND: The Medtrac MDI Chronolog is an electronic device for monitoring adherence to metered-dose inhalers. It replaces previous models of the Nebulizer Chronolog and uses a different mechanism of recording actuations. OBJECTIVE: This study was carried out to determine whether the new model can accurately record and report the date, time, and number of metered-dose inhaler actuations. METHODS: Four canisters of beclomethasone (Beclovent) were discharged through four Chronologs with fresh batteries at a rate of 1, 2, 4, or 8 times twice daily for 7 days. Four additional canisters were used as controls and discharged simultaneously through the standard actuator. The weight of all canisters and Chronolog battery voltage were measured before and at the end of the 7-day experiment. The data retrieved from the Chronologs were compared with the information recorded manually during each discharge. RESULTS: The loss in canister weight was consistent for the number of puffs discharged from all four Chronolog units and controls. However, the accuracy of the Chronologs in recording the number of actuations varied between 50% and 100%. The largest amount of data loss occurred with the unit used to discharge 8 puffs and was associated with a dead battery at the end of the 7-day trial. For actuations that were retrievable, the Chronologs accurately recorded the date and time. CONCLUSIONS: Unexpected battery voltage drain and other mechanical problems that we encountered may cause data loss and the false appearance of missed doses. Thus the units that we tested were not sufficiently reliable to monitor patient adherence. PMID- 9338543 TI - Allergic sensitization increases airway reactivity in guinea pigs with respiratory syncytial virus bronchiolitis. AB - BACKGROUND: Respiratory syncytial virus (RSV) causes acute bronchiolitis in children and has been implicated in the pathogenesis of recurrent wheezing and asthma. However, few children exposed to RSV experience acute bronchiolitis or its sequelae, suggesting a subgroup of susceptible children. An allergic diathesis may predispose children to subsequent airway disease. OBJECTIVE: This study was carried out to determine whether a preexisting allergic state, induced by repeated inhalational exposures to ovalbumin, potentiates nonspecific airway responsiveness to acetylcholine and increases airway inflammation during acute RSV bronchiolitis in guinea pigs. METHODS: Forty guinea pigs were randomized into four groups: nonsensitized, noninfected (ovalbumin-, RSV-); sensitized, noninfected (ovalbumin+, RSV-); nonsensitized, infected (ovalbumin-, RSV+); sensitized, infected (ovalbumin+, RSV+). Depending on grouping, animals were exposed to either repeated aerosols of ovalbumin or saline solution and were subsequently inoculated with either human RSV or uninfected culture medium. Six days after inoculation, animals underwent acetylcholine challenge, and lung specimens were prepared for histologic scoring of airway inflammation. RESULTS: Maximal increases in pulmonary resistance (centimeters of water per milliliter per second) to acetylcholine were greater for RSV alone (12.4 +/- 3.9) and ovalbumin alone (13.7 +/- 3.9) compared with controls (4.3 +/- 1.1), but significantly greater increases occurred in ovalbumin+, RSV+ animals (34.0 +/- 11.0). These ovalbumin+, RSV+ animals demonstrated the combined histologic changes noted with RSV alone and ovalbumin alone including airway epithelial necrosis, mononuclear and granulocyte infiltrates, airway wall edema, hyperplasia of bronchus-associated lymphoid tissue, and goblet cell metaplasia. CONCLUSION: Prior allergic sensitization potentiates the physiologic and structural changes induced by acute RSV bronchiolitis. These results suggest that an allergic diathesis may increase the severity of RSV infections in children. PMID- 9338544 TI - Cultured nasal polyps from nonatopic and atopic patients release RANTES spontaneously and after stimulation with phytohemagglutinin. AB - BACKGROUND: Eosinophil infiltration of tissue is a hallmark of nasal polyposis in both nonatopic and atopic patients. These cells are thought to play a key role in the nasal polyp inflammatory process. OBJECTIVE: The objective of this study was to investigate whether cultured nasal polyps derived from nonatopic and atopic patients release RANTES both spontaneously and after phytohemagglutinin (PHA) stimulation. METHODS: Nasal polyps were obtained from 12 subjects (6 nonatopic and 6 atopic), cut into 2 to 3 mm large specimens, and cultured for 48 hours with or without PHA. RANTES was measured in the culture supernatant by ELISA (R&D Systems, U.K.). RESULTS: Immunoreactive RANTES was found to be present in the culture supernatant of nasal polyps derived from both nonatopic and atopic patients with no difference between the two groups (median: 3.8 vs 2.9 pg/mg/ml). On incubation with PHA, nasal polyps from both nonatopic and atopic patients released sevenfold and 11-fold greater amounts of RANTES than unstimulated samples. As determined by immunohistochemistry, RANTES was localized to the vascular endothelium in nasal polyps from both groups of patients. CONCLUSIONS: This study demonstrates that cultured nasal polyps derived from both nonatopic and atopic patients release RANTES spontaneously and after PHA stimulation. This observation and the finding that RANTES is present in nasal polyp endothelial cells suggest that this chemokine may be an important mediator of eosinophil and lymphocyte recruitment in both nonatopic and atopic nasal polyposis. PMID- 9338545 TI - Nasal provocation with allergen induces a secondary serum IgE antibody response. AB - The study of the IgE response to seasonal antigen exposure is limited by its occurrence once a year and by the variability of patient exposure to pollens. To overcome these problems, we investigated whether nasal challenge with antigen causes an increase in serum anti-ragweed IgE levels. We challenged individuals with ragweed allergy intranasally with nanogram quantities of ragweed antigen extract and measured their serum anti-ragweed IgE levels before and at weekly intervals after challenge. In a series of studies we found that there was a reproducible rise in antigen-specific serum IgE levels beginning the first week after challenge that plateaued at about 180% of baseline levels during the fourth week and remained elevated for 8 weeks. Not all individuals showed this response. The magnitude of the allergen-specific IgE response to nasal challenge appeared to be greater than the response to seasonal exposure. Treatment with intranasal beclomethasone before challenge did not affect the response. The results demonstrate a human in vivo model for the study of the antigen-specific secondary IgE response to allergen. PMID- 9338546 TI - The inhibitory effects of topically active glucocorticoids on IL-4, IL-5, and interferon-gamma production by cultured primary CD4+ T cells. AB - This study was conducted to directly compare the in vitro efficacy and potency of several glucocorticoids in inhibiting T-cell cytokine production. The glucocorticoids tested were fluticasone propionate, budesonide, triamcinolone acetonide, and beclomethasone dipropionate, which are currently inhaled therapies for the treatment of allergic airway disease. Also used were betamethasone phosphate and the newly developed mometasone furorate. With a novel cell culture system, purified peripheral blood CD4+ T cells from normal donors were stimulated with immobilized anti-CD3 and soluble anti-CD28 monoclonal antibodies to induce high levels of IL-4, IL-5, and interferon-gamma. By cell sorting, it was found that the IL-5 produced originated from memory cells, whereas both memory and naive cells produced interferon-gamma. Mometasone and fluticasone inhibited IL-5 and IL-4 similarly (mometasone IL-5 inhibitory concentration of 50% = 0.27 +/- 0.1 nmol/L and IL-4 = 0.19 +/- 0.08 nmol/L). For both cytokines, the results indicate that mometasone and fluticasone were more potent than beclomethasone, triamcinolone, budesonide, and betamethasone. Of clinical importance is the finding that all steroids demonstrated less efficacy versus interferon-gamma than IL-4 and IL-5. Glucocorticoid reduction of Th2 cytokines with lesser effects on interferon-gamma would serve to reverse the exaggerated Th2 response that contributes to pathophysiology observed in allergic disease. Therefore the use of topically active glucocorticoids with low systemic bioactivity for the treatment of allergic inflammation may be particularly effective in modulating the cytokine activity that is an important component of the allergic response. PMID- 9338547 TI - Platelet activating factor compromises airway epithelial defense functions. AB - BACKGROUND: The mechanism of disruption of the epithelial defense function observed in asthmatic airways is considered to be largely the result of mediators involved in allergic responses. Platelet activating factor (PAF) might be a key mediator involved in this mechanism. OBJECTIVE: This study was designed to determine whether PAF is capable of compromising the epithelial defense functions, such as tight junctional barriers and the mucociliary system. METHODS: A total of 120 healthy rabbits were used. Twenty of them were used as normal controls. Eighty rabbits were treated with inhalation of 10 ml of PAF (200 microg/ml), and 20 animals were used for the examination of epithelial defense functions of the trachea at 1, 10, 20, and 30 days after inhalation of PAF. Epithelial defense functions of the trachea were evaluated by ciliary activity, mucociliary transport velocity, epithelial permeability to fluorescein isothiocyanate dextrans (70,000 d), and electron microscopy of the epithelial structure. RESULTS: PAF inhalation induced a significant decrease in ciliary activity and mucociliary transport velocity, which persisted for up to 20 days. PAF inhalation also caused a significant 7.4-fold increase in epithelial permeability to fluorescein isothiocyanate dextrans at I and 10 days. This increased epithelial permeability returned to the normal level 20 days after PAF inhalation. However, electron microscopy demonstrated no apparent evidence of epithelial shedding. CONCLUSIONS: PAF-induced prolonged dysfunction of both the epithelial junctional barrier and the mucociliary system may allow enhanced entry of allergen molecules, as well as bronchoactive agonists to the airway parenchyma and may also significantly contribute to an increased airway responsiveness. PMID- 9338548 TI - Phosphodiesterase profiles of highly purified human peripheral blood leukocyte populations from normal and atopic individuals: a comparative study. AB - BACKGROUND: Several previous reports have suggested an increased activity of cAMP phosphodiesterases (PDEs) and a higher sensitivity of these enzymes toward PDE inhibitors in leukocytes of patients suffering from atopic dermatitis. OBJECTIVE: The purpose of the present study was to comprehensively analyze and compare the PDE expression and activity profile of highly purified populations of leukocytes from normal and atopic blood donors. In addition, the influence of PDE inhibitors on function of leukocytes from normal and atopic individuals was investigated. METHODS: Density gradient centrifugation, elutriation, and magnetic cell sorting techniques were used to purify eosinophils, monocytes, and B and T lymphocytes from peripheral human blood. Complementary DNA-polymerase chain reaction was used to analyze PDE4 subtype messenger RNA (mRNA) expression levels in addition to PDE isoenzyme activities. PDE4 inhibitor sensitivity was determined in monocyte homogenates from both groups. Functionally, suppression of lipopolysaccharide induced synthesis of tumor necrosis factor-alpha in monocytes as well as phytohemagglutinin-induced T cell proliferation in peripheral blood mononuclear cell fractions by PDE4 and PDE3/4 inhibitors was compared. RESULTS: Identical PDE activities and mRNA expression profiles were found in all cells from normal and atopic donors except that there was an increase in the mRNA levels of PDE4A and PDE4B2 in atopic T cells, which was, however, not reflected in overall PDE4A activity. In addition, no differences in sensitivity of the functional responses to PDE inhibitors were noted. The mixed PDE3/4 inhibitor zardaverine was a more potent inhibitor of T cell proliferation than rolipram, a selective PDE4 inhibitor. CONCLUSION: No evidence for alterations of PDE activities in atopy is provided by our findings. PMID- 9338549 TI - Delayed eosinophil programmed cell death in vitro: a common feature of inhalant allergy and extrinsic and intrinsic atopic dermatitis. AB - The present studies were undertaken to characterize the potential role of eosinophil programmed cell death (PCD) in atopic diseases. Peripheral blood eosinophil PCD was found to be delayed in inhalant allergy (p < 0.05) and delayed to an even greater extent in atopic dermatitis (AD) (p < 0.0001) when compared to nonatopic subjects. There was no difference in the occurrence of PCD between the extrinsic and the intrinsic type of AD, pointing to a secondary role of specific sensitization. Blockade of eosinophil PCD was not responsible for peripheral blood eosinophilia, because we found no obvious relationship of eosinophil survival to blood eosinophil count. Eosinophil supernatants of more patients with AD than of patients with inhalant allergy dose-dependently inhibited PCD in nonatopic eosinophils, and it was shown that this effect was possibly due to autocrine production of granulocyte-macrophage-colony stimulating factor, probably IL-5. Eosinophil expression of CD95 (Fas antigen) did not change over time in culture and was not modulated by cytokines prolonging eosinophil survival. In contrast, IL-3, IL-5, and granulocyte-macrophage-colony stimulating factor caused an upregulated expression of CD69. However, in AD, CD69 on eosinophils was upregulated without the need of exogenous growth factor or factors over time in culture, thus confirming an autocrine production of proeosinophilic cytokines. In conclusion, our data clearly indicate that eosinophil PCD is markedly delayed in the so-called atopic diseases irrespective of allergen sensitization and suggest that this effect is mediated by the autocrine production of growth factors by eosinophils. PMID- 9338550 TI - Immunoreactive endothelin in bronchial biopsy specimens: increased expression in asthma and modulation by corticosteroid therapy. AB - BACKGROUND: The human endothelin (ET) family comprises three 21-amino-acid peptides, which are potent bronchoconstrictors and have a number of other biologic properties relevant to the pathophysiology of asthma. OBJECTIVE: We sought to compare the expression of immunoreactive ET in bronchial biopsy specimens from subjects with asthma treated only with inhaled beta2-agonists, subjects with asthma treated with beta2-agonists and corticosteroids, and control subjects without asthma. METHODS: Biopsy specimens were obtained by fiberoptic bronchoscopy and stained immunohistochemically with a specific ET antiserum. Epithelial ET expression was quantitated by using a computer-assisted system of image analysis. Numbers of inflammatory cells and depth of subepithelial collagen deposition were also determined. RESULTS: Immunoreactive ET was principally localized in the airway epithelium. The proportion of epithelium immunostained was significantly increased in the subjects with asthma not treated with steroids (35.4% +/- 3.8%) compared with that of both the control subjects (16.2% +/- 1.9%, p < 0.0001) and the subjects with asthma treated with steroids (14.3% +/- 2.0%, p < 0.0001). The last two groups did not differ significantly from one another. There were no significant correlations between ET expression and either physiologic parameters or indexes of airway inflammation and remodeling. CONCLUSION: Bronchial epithelial expression of immunoreactive ET is increased in subjects with asthma receiving treatment only with beta2-agonists but not in subjects with asthma also receiving corticosteroid therapy. These findings are consistent with the hypothesis that ET is implicated in the pathophysiology of asthma. PMID- 9338551 TI - Suppression of plasma neutrophil elastase-alpha1-anti-proteinase complex by adequate replacement therapy in a patient with acquired hypogammaglobulinemia. PMID- 9338552 TI - Cytokine production in transient hypogammaglobulinemia and isolated IgA deficiency. AB - BACKGROUND: Transient hypogammaglobulinemia of infancy and isolated IgA deficiency are characterized by normal numbers of circulating B lymphocytes. It is likely that no single abnormality, but rather different factors, may be relevant for the delayed onset of IgG synthesis in transient hypogammaglobulinemia or for the differentiation defect of B cells in IgA deficiency. These factors may include defective production of cytokines or an abnormal response of B cells to various mediators. Alternatively, some cytokines may act as inhibitory factors of B-cell function. METHODS: The ability of peripheral blood mononuclear cells from children with proved or probable transient hypogammaglobulinemia (30 patients) and IgA deficiency (15 patients) to secrete several cytokines on stimulation with phytohemagglutinin in vitro was analyzed. RESULTS: An enhanced production of tumor necrosis factor (TNF)-alpha, TNF-beta, and IL-10 was observed in transient hypogammaglobulinemia; whereas secretion of IL-1, IL-4, and IL-6 was essentially similar in the control and patient groups. Increased frequency of mononuclear cells secreting TNF-alpha was seen in the patient groups. Apart from elevated production of TNF-alpha, no other abnormalities in cytokine synthesis in selective IgA deficiency were observed. In vitro observations showed that exogenously added TNF-alpha and TNF-beta inhibited IgG and IgA secretion by pokeweed mitogen-stimulated mononuclear cells. During follow-up of 10 children, normalization of serum IgG level was associated with a decrease in previously elevated TNF-alpha and TNF-beta production, but IL-10 production remained unchanged. CONCLUSION: These results suggest that TNF may be involved in the regulation of IgG and IgA production and can be associated with an arrest of IgG and IgA switch of B cells in hypogammaglobulinemia. The balance between TNF and IL-10 may be important for the normal development of IgG secreting B cells. PMID- 9338553 TI - Total IgE serum levels correlate with sinus mucosal thickness on computerized tomography scans. AB - BACKGROUND: Sinusitis involves an inflammatory response with similarities to asthma. OBJECTIVE: We sought to determine whether a correlation exists between total IgE and the sinus mucosal thickening as assessed by computed tomography (CT) scans. METHODS: We screened the charts of 300 otolaryngology patients who had total and specific serum IgE determinations in 1994 and from this group selected all patients who had also undergone a sinus CT scan close to the time of serum evaluation (n = 86). Severity of disease on CT scan was graded by two investigators blinded to IgE levels. RESULTS: There was a significant positive correlation between severity of disease on CT scans and IgE (r[s] = 0.37, p = 0.0007). Furthermore, patients with a more advanced disease stage had higher IgE levels. There was also a positive correlation between severity of disease and the sum of specific IgE grades (r[s] = 0.29, p = 0.007). CONCLUSION: The data suggest that IgE levels or a linked genetic parameter may contribute to the mucosal inflammation in the paranasal sinuses. PMID- 9338554 TI - Expression of IL-16 in allergen-induced late-phase nasal responses and relation to topical glucocorticosteroid treatment. AB - Allergen-induced late nasal responses (LNRs) are associated with a cellular infiltrate in which CD4+ cells are prominent. These cells have been shown to be the major cellular source of Th2-type cytokines. Mechanisms responsible for the local accumulation of CD4+ cells in the nasal mucosa after allergen exposure are unclear. IL-16 is a potent chemoattractant for CD4+ cells in vitro and may play a significant role in recruiting CD4+ cells in LNRs. We investigated the expression of IL-16 messenger RNA and immunoreactivity in nasal biopsy specimens from 17 subjects with allergic rhinitis. A biopsy specimen of the nasal inferior turbinate was obtained before and 24 hours after local nasal provocation with grass pollen extract after 6 weeks of treatment with either topical fluticasone propionate (n = 9) or placebo (n = 8) nasal spray twice daily. IL-16 mRNA positive cells and IL-16-immunoreactive cells were identified in both the epithelium and the subepithelial tissue at baseline. Within the placebo-treated group, the numbers of epithelial and subepithelial IL-16 mRNA-positive cells and IL-16-immunoreactive cells were significantly increased 24 hours after challenge compared with baseline (p < 0.001). Topical glucocorticoid therapy resulted in a decrease in allergen-induced epithelial immunoreactive cells and subepithelial IL 16 mRNA-positive cells. The numbers of CD4+ cells increased after antigen challenge compared with baseline (p < 0.05), and this increase was inhibited by glucocorticoid treatment. There were significant correlations between epithelial and subepithelial IL-16 immunoreactivity and CD4+ cell infiltration after antigen challenge. The upregulation of IL-16 expression in allergic nasal mucosa after antigen challenge may have critical implications in the accumulation of CD4+ cells in response to antigen exposure. Steroid-mediated inhibition of IL-16 may be partly responsible for the decrease in local CD4+ cells after topical glucocorticoid therapy. PMID- 9338555 TI - Asthma caused by occupational exposure to pectin. PMID- 9338556 TI - Allergy to chicken meat without sensitization to egg proteins: a case report. PMID- 9338557 TI - The effects of temperature and buffer on the extraction of Der p 1 from dust. PMID- 9338558 TI - Habit reversal technique and eczema. PMID- 9338579 TI - Registries of immunodeficiency patients and mutations. AB - Immunodeficiencies form a distinct group of human hereditary diseases with several rare disorders. During recent years, information has been collected concerning immunodeficiency patients and mutations causing disorders. The large European (ESID) registry contains clinical data for some 7,000 patients. At present, international mutation databases have information for > 1,000 immunodeficiency patients, including X-linked chronic granulomatous disease (XCGD), Wiskott-Aldrich syndrome (WAS), and X-linked thrombocytopenia (XLT), X linked hyper-IgM syndrome (XHIM), X-linked agammaglobulinemia (XLA), and X-linked severe combined immunodeficiency (XSCID). The databases are available on Internet. The mutation spectra of patients in these registries were compared. Mutational hotspots were found in CpG dinucleotides with a preference for selected flanking bases. PMID- 9338580 TI - Molecular genetics of Krabbe disease (globoid cell leukodystrophy): diagnostic and clinical implications. AB - Galactocerebrosidase (GALC) is a lysosomal beta-galactosidase responsible for the hydrolysis of the galactosyl moiety from several galactolipids, including galactosylceramide and psychosine. The deficiency of this enzyme results in the autosomal recessive disorder called Krabbe disease. It is also called globoid cell leukodystrophy (GLD), because of the characteristic storage cells found around cerebral blood vessels in the white matter of affected human patients and animal models. Although most patients present with clinical symptoms before 6 months of age, older patients, including adults, have been diagnosed by their severe deficiency of GALC activity. More than 40 mutations have been identified in patients with all clinical types of GLD. While some mutations clearly result in the infantile type if found homozygous or with another severe mutation, it is difficult to predict the phenotype of novel mutations or when mutations are found in the heterozygous state. A high incidence of polymorphic changes on apparent disease-causing alleles also complicates the interpretation of the effects of mutations. The detection of mutations has greatly improved carrier identification among family members and will permit preimplantation diagnosis for some families. The molecular characterization of the naturally occurring mouse, dog, and monkey models will permit their use in trials to evaluate different modes of therapy. PMID- 9338581 TI - Mutation screening of all 65 exons of the fibrillin-1 gene in 60 patients with Marfan syndrome: report of 12 novel mutations. AB - Mutations in the fibrillin-1 gene on chromosome 15q21.1 have been found to cause Marfan syndrome, a dominantly inherited disorder characterised by clinically variable skeletal, ocular, and cardiovascular abnormalities. In this study we screened all 65 exons of the fibrillin-1 gene in 20 Marfan syndrome families where at least two affected individuals were characterised and available for analysis, another 30 families with only one affected member available for analysis, and in 10 sporadic cases. In large well-characterised families with more than four affected individuals, the detection rate for mutations rose to 78% (7/9), in families with either two or three affected members 27% (3/11). In families where only one affected family member was available, the mutation detection rate was 17% (5/30), and in sporadic cases it was 20% (2/10). In addition, we found eight neutral polymorphisms. Twelve of the 17 disease-causing mutations identified have not been previously described, thus raising the total number of different fibrillin-1 mutations reported to 85 in 94 unrelated cases. PMID- 9338582 TI - Evidence for founder effect of the Glu104Asp substitution and identification of new mutations in triosephosphate isomerase deficiency. AB - Triosephosphate isomerase (TPI) deficiency is an autosomal recessive disorder of glycolysis characterized by multisystem disease and lethality in early childhood. Among seven unrelated Northern European kindreds with clinical TPI deficiency studied, a single missense mutation at codon 104 (GAG;Glu-->GAC;Asp) predominated, accounting for 11/14 (79%) mutant alleles. In three families molecular analysis revealed compound heterozygosity for Glu104Asp and novel missense mutations. In two cases the second mutation was a Cys to Tyr substitution at codon 41 (TGT-->TAT) and in one an Ile to Val substitution at codon 170(ATT-->GTT). The origin of the Glu104Asp mutation was defined by haplotype analysis using a novel G/A polymorphism at nucleotide 2898 of the TPI gene. Cosegregation of the low frequency 2898A allele with the G-->C base change at nucleotide 315 supports a single origin for the Glu104Asp mutation in a common ancestor. PMID- 9338583 TI - Tay-Sachs disease and HEXA mutations among Moroccan Jews. AB - Moroccan Jewry (N>750,000) is the only non-Ashkenazi Jewish community in which Tay-Sachs disease (TSD) is not extremely rare. Previous studies among Moroccan Jewish TSD families identified three HEXA mutations. In this study, extended to enzyme-defined and new obilgate TSD carriers, we found four additional mutations. One of them is a novel, IVS5-2(A-->G) substitution, resulting in exon skipping, and it was found only among enzyme-defined carriers. The seven HEXA identified mutations among Moroccan Jews are: deltaF(304/305), R170Q, IVS-2(A-->G), Y180X, E482K, 1278+TATC, and IVS12+1(G-->C). Their respective distribution among 51 unrelated enzyme-defined and obligate carriers is 22:19:6:1:1:1:1. The mutation(s) remain unknown in only three enzyme-defined carriers. Five of the seven Moroccan mutations, including the three most common ones, were not found among Ashkenazi Jews. Compared with the much larger and relatively homogeneous Ashkenazi population, the finding among Moroccan Jews probably reflects their much longer history. PMID- 9338584 TI - Adult vitelliform macular dystrophy is frequently associated with mutations in the peripherin/RDS gene. AB - Mutations in the peripherin/RDS gene, which encodes a photoreceptor-specific membrane glycoprotein, have been identified in a variety of retinal phenotypes. However, the mechanisms by which specific mutations in this gene can cause typical features of retinal dystrophies clinically as distinct as retinitis pigmentosa or macular degeneration are still unknown. Recently, a single case of adult vitelliform macular dystrophy (AVMD) has been associated with a Y258Stop mutation. To assess the frequency of peripherin/RDS mutations in the clinically heterogeneous group of AVMD, we analyzed the entire coding region of the gene in 28 unrelated patients. We identified five novel mutations including two presumed null allele mutations. Thus, our results demonstrate that a significant portion of AVMD patients (18%) carry point mutations in peripherin/RDS, suggesting that this gene is frequently involved in the pathogenesis of this macular disorder. In addition, this study shows that the variable phenotypes in AVMD are due, at least in part, to genetic heterogeneity and are likely to be caused by mutations in disease genes thus far unknown. PMID- 9338585 TI - Characterization of a deletion mutation involving exons 3-7 of the WASP gene detected in a patient with Wiskott-Aldrich syndrome. AB - A case of Wiskott-Aldrich syndrome (WAS) suspected to have a deletion mutation in the WAS protein (WASP) gene had previously been reported (Ariga et al., 1997). Genomic polymerase chain reaction (PCR) suggested that exons 3-7 of the WASP gene were included in the deletion. Present Southern blot studies confirm that the deletion is approximately 2.0 kb in length, involving exons 3-7 and seemed to have been created by the fusion of introns 2 and 7. To characterize the deletion mutation in detail, we analyzed the PCR-amplified fragments of introns 2 and 7 from normal individuals and the fragment suspected of including the deletion junction from the patient. Sequencing of the patient fragment revealed that the deletion mutation involving exons 3-7 of the WASP gene did, indeed, result from the fusion of introns 2 and 7. PMID- 9338586 TI - Confirmation of homozygosity for a single nucleotide substitution mutation in a Cockayne syndrome patient using monoallelic mutation analysis in somatic cell hybrids. AB - The identification of individuals homozygous for a specific mutation offers advantages for the elucidation of molecular mechanisms of hereditary disease states. Cockayne syndrome is a rare autosomal recessive disorder, the molecular basis of which is complicated by significant genetic and clinical heterogeneity. The genes associated with both genetic complementation groups, CSA and CS-B, have been identified. We have previously identified a number of CSA mutations, including a single base substitution that introduces a stop codon (322Tyr-->Stop) mutation in the C-terminal region for at least one allele of the CSA gene in a severely affected patient. We now present data confirming the existence of homozygosity in this patient using a strategy with general applicability. Somatic cell hybrids were established by fusing patient cells with mouse A9 cells. Screening with chromosome 5 specific polymorphic markers facilitated identification of hybrid clones bearing only one of the distinct CSA alleles. Sequencing of a portion of the human CSA gene in a subset of these hybrids permitted monoallelic mutation analysis and confirmed the presence of the 322Tyr- >Stop mutation in both alleles. PMID- 9338587 TI - Quantitative enriched PCR (QEPCR), a highly sensitive method for detection of K ras oncogene mutation. AB - We have developed a rapid and highly sensitive method for the detection of mutant K-ras codon 12 allele in the presence of 10(5) copies of the wild-type alleles. This sensitivity is achieved by selective amplification of mutant K-ras sequences, using a two-stage procedure with modified primers. In the first stage, primers consist of K-ras sequences in the 3' portion and polyomavirus sequence (to minimize homology with human genome) on the 5' portion. The 3' portion also consists of mismatch sequence that generates an MvaI site in normal, but not mutant, K-ras codon 12 alleles. Thus, following the first round of 20 cycles, restriction enzyme cleavage is carried out to selectively digest normal K-ras codon 12 alleles. To enrich mutant alleles, a second amplification is performed using tail primers that recognize the polyoma, but not human sequences. This design ensures that in the second amplification only mutant alleles that were pre amplified in the first round would serve as template for this reaction. Ethidium bromide-stained polyacrylamide gel electrophoresis (PAGE) of second-stage PCR product that has been digested with MvaI is used to monitor the presence of mutant alleles, detected at sensitivity of 1/10(5). This technique offers high sensitive detection of mutant K-ras alleles using a new concept of tail-primer design and is likely to assist in identifying patients at risk to develop pancreatic, colon, or lung cancer, which harbor high incidence of mutant ras alleles. PMID- 9338588 TI - P1148A in fibrillin-1 is not a mutation leading to Shprintzen-Goldberg syndrome. PMID- 9338589 TI - Nuclear envelope dynamics during male pronuclear development. AB - Upon fertilization, the sperm nucleus undergoes reactivation. The poreless sperm nuclear envelope is replaced by a functional male pronuclear envelope and the highly compact male chromatin decondenses. Here some recent evidence is examined: that disassembly of the sperm lamina is required for chromatin decondensation, that remnant portions of the sperm nuclear envelope target the binding of egg membrane vesicles that form the male pronuclear envelope, that functional male pronuclear envelopes containing lamin B receptor assemble prior to lamin import and lamina formation, and that lamina assembly drives male pronuclear swelling. Several unresolved issues are discussed. PMID- 9338590 TI - Identification of sperm plasma membrane proteins exhibiting binding affinity for the ascidian egg coat. AB - In the initial stage of ascidian fertilization sequential sperm-egg coat interactions assure successful species-specific fertilization. Sperm recognize, bind to, and then penetrate the egg investment that consists of follicle cells (FC) and an acellular vitelline coat (VC). To identify plasma proteins that recognize the egg coat, a membrane fraction was prepared from Phallusia mammillata sperm using nitrogen cavitation followed by three centrifugation steps. The purity of the membrane fractions was assessed by transmission electron microscopy and marker enzymes. Comparison of the electrophoretic pattern of sperm extracellular membrane domains labeled by radio-iodination or biotinylation and recorded by autoradiography or enhanced chemiluminescence, respectively, showed the non-radioactive procedure to be a convenient and efficient method. Isolated sperm membrane components were found to inhibit fertilization in a concentration dependent manner and to bind mainly to the FC. Eggs were used as an affinity matrix to determine which of the solubilized sperm membrane proteins possess egg binding activity. Three biotinylated proteins (66 kDa, 120 kDa and 140 kDa) were found to bind to the VC. Assays probing heterospecific binding to Ascidia mentula eggs revealed that the 120 kDa protein possesses species-specific binding activity. Thus, the current data suggest the 120 kDa sperm membrane protein as a candidate adhesion molecule with a possible role in gamete binding and species specific recognition in P. mammillata. PMID- 9338591 TI - Alterations in sarcomere structure, collagen organization, mitochondrial activity, and protein metabolism in the avian low score normal muscle weakness. AB - Skeletal muscle fibers are surrounded by an extracellular matrix. The extracellular matrix is composed of glycoproteins, collagen, and proteoglycans. Proteoglycans have been suggested to play an important functional role in tissue differentiation. However, an understanding of how the extracellular matrix affects skeletal muscle development and function is largely unknown. In the avian genetic muscle weakness, low score normal (LSN), a late embryonic increase in the expression of decorin is followed by a subsequent increase in collagen crosslinking. The sarcomere organization, collagen fibril diameter and organization were investigated using transmission electron microscopy. Measurements were made at 20 days of embryonic development and 6 weeks posthatch. These studies showed changes in sarcomere organization and deterioration of muscle fibril structure in the LSN pectoral muscle. In vitro satellite cell cultures were developed and assayed for mitochondrial activity, and protein synthesis and degradation. In these analyses, mitochondrial activity from LSN satellite cells was significantly higher than those from normal pectoral muscle satellite cells. Protein synthesis rates between the normal and LSN satellite cell-derived myotubes were similar, but protein degradation rates were higher in the LSN cultures. Based on the reported functions of decorin as a regulator of cell proliferation and collagen fibril organization, it is possible that the late embryonic increase in decorin may be influencing the alterations in LSN sarcomere and collagen organization. PMID- 9338592 TI - The Na+,K+-ATPase alpha subunit requires gastrulation in the Xenopus embryo. AB - Na+,K+-ATPase participates in reabsorption of ions and water and produces an electrochemical gradient between the intra- and extracellular spaces across the cell membrane. It also plays an important role in many developmental phenomena such as a blastocoele formation and neural formation. To elucidate the expression pattern of Na+,K+-ATPase in the Xenopus embryo, the spatial expression patterns of the Na+,K+-ATPase alpha subunit were studied in a normal embryo by whole-mount in situ hybridization. These transcripts were localized around the dorsal blastopore at the gastrula stage, in the neural tube at the neurula stage, and then in the pronephros and cloaca at the tail-bud stage. To study the function of Na+,K+-ATPase in embryogenesis after mid-blastula transition, the expression of the Na+,K+-ATPase alpha subunit was inhibited by the injection of specific antisense RNA. Embryos injected with Na+,K+-ATPase antisense RNA showed inhibition of gastrulation. When antisense RNA was injected into the dorsal blastomeres, head differentiation was markedly inhibited. These results suggest that this transcript plays an important role during gastrulation and head differentiation. PMID- 9338593 TI - Formation of the branching pattern of blood vessels in the wall of the avian yolk sac studied by a computer simulation. AB - The present study was performed to provide data to support the notion previously believed but not proved experimentally or theoretically, that blood vessels are formed by the selection of capillaries in the network. In an attempt to understand the mechanism of formation of blood vessel branching structures, the transformation of a capillary network to a branching system in the wall of quail yolk sac was successively recorded by a series of photographs, and a computer simulation was carried out for the process of in vivo vascularization based on the photographs. The simulation demonstrated that a positive feedback system participated in the formation of a branching structure. That is, vessels which had been much used were enlarged, whereas less used vessels were reduced in their size and finally extinguished. The enlarged vessels became major components of the branching system. As the body of an embryo grew, it was observed that polygonal capillary networks enlarged, which led each polygon of the network to divide into a few finer polygons. Then, some of the capillary vessels were again selected and formed a branching system. This process repeated during the body growth, indicating that the vascular system developed adaptively to the body growth. A region where the growth was fast, received much blood flow and produced finer networks of capillaries. Thus, it was experimentally demonstrated for the first time that capillaries in the network are successively selected by a positive feedback mechanism and form blood vessels. PMID- 9338594 TI - gp49B1, an inhibitory signaling receptor gene of hematopoietic cells, is induced by leukemia inhibitory factor in the uterine endometrium just before implantation. AB - Leukemia inhibitory factor (LIF) is a cytokine involved in hematopoiesis, neuropoiesis, and embryogenesis. Transcriptional activation of various genes occurs subsequent to LIF signal transduction in its target cells. Using the mRNA differential display method, a LIF-inducible gene was isolated from LIF stimulated M1 murine myeloid leukemia cells. By DNA sequencing, this gene turned out to be gp49B1, which has been reported as an inhibitory signaling receptor to attenuate mast cell activation. Because gp49B1 expression was limited to the uterus of a pregnant mouse, its uterine expression was examined especially in relation to LIF expression during pregnancy. gp49B1 was expressed specifically on day 4.0 of pregnancy, as was LIF, and the site of the most abundant expression of LIF and gp49B1 mRNA was the luminal epithelium of the uterine endometrium. These findings suggest that the gp49B1 expression in the uterine endometrium is induced just before implantation by paracrine and/or autocrine effects of LIF. Considering its function as an inhibitory signaling receptor on mast cells, a possible role for gp49B1 on the surface of the uterine endometrium as an immunoreceptor that allows blastocyst attachment is proposed. PMID- 9338595 TI - Reciprocal transplantation of wing discs between a wing deficient mutant (fl) and wild type of the silkworm, Bombyx mori. AB - The wingless mutant flugellos (fl) of the silkworm lacks all four wings. Although wing discs of the fl seem to develop normally until the fourth larval instar, wing morphogenesis stops after the fourth larval ecdysis, probably caused by aberrant expression of an unidentified factor, referred to as fl. To characterize factor fl, the wing discs dissected from the wild-type (WT) and fl larvae were transplanted into other larvae and developmental changes of the discs were examined. When the wing disc from a WT larva was transplanted into another WT larva and allowed to grow until emergence, a small wing appeared that was covered with scales. Thus, the transplanted wing discs can develop autonomously, form scales and evert from adult skin. The WT wing discs transplanted into the fl larvae also developed at a high rate. However, the fl wing discs transplanted into the WT larvae did not develop during the larval to pupal developmental stages. These data suggest that the fl gene product (factor fl) works in the wing disc cells during wing morphogenesis. Its function cannot be complemented by hemolymph in the WT larva. It is also implied that the level of humoral factors and hormones required for wing morphogenesis are normally maintained in the fl larva. PMID- 9338596 TI - Development of the competence of bovine oocytes to release cortical granules and block polyspermy after meiotic maturation. AB - Bovine immature oocytes do not have the ability to block polyspermic penetration. The present study was conducted to determine whether this is correlated to cortical granule (CG) distribution and the competence of oocytes to release CG upon sperm penetration, and whether the ability of bovine oocytes to release CG develops during in vitro maturation. Fluorescein isothiocyanate-conjugated Lens culinaris agglutinin was used for detecting CG in immature and mature oocytes before and after sperm penetration and electric stimulation. The labeled oocytes were examined with laser confocal and fluorescent microscopes. The results show that CG exist as clusters in all immature oocytes. The CG were not released from immature oocytes exposed to electric pulse or penetrated by spermatozoa, resulting in 94% of oocytes being polyspermic. When immature oocytes were cultured for 22 h in vitro, 81% extruded the first polar body and reached metaphase II. In mature oocytes, 25% of oocytes showed CG clusters, 42% and 33% of oocytes showed partial and complete CG dispersion, respectively. When mature oocytes were inseminated in vitro, only 15% of oocytes were polyspermic. Cortical granule exocytosis occurred in 97% of oocytes after sperm penetration and 84% of oocytes released all of the CG 18 h after insemination. Electric pulse induced all of the mature oocytes to release CG but only 55% released all of their CG 18 h post stimulation. These results indicate that polyspermy in immature bovine oocytes is the result of the complete failure of the oocyte to release CG after sperm penetration. Bovine oocytes became competent to release CG by sperm penetration and electric stimulation after meiotic maturation. These results provide evidence that CG exocytosis plays an important role(s) in the establishment of the block to polyspermy in bovine oocytes. PMID- 9338597 TI - Analysis of mRNA levels for developmentally regulated prespore specific glutamine synthetase in Dictyostelium discoideum. AB - The enzyme glutamine synthetase (GS) of Dictyostelium discoideum is developmentally regulated, preferentially localized in prespore cells and is likely to play an important role in controlling the levels of ammonia, a known morphogen, in this organism. To further investigate the regulation of GS, a portion of the GS gene was isolated and used as a probe to examine the changes in GS mRNA throughout development and the level of GS mRNA in the two precursor cell types. The amino acid sequence of the cloned DNA fragment isolated is highly homologous to other eukaryotic GS genes. DNA blot analysis demonstrated that the GS gene exists as a single copy in D. discoideum. Analysis of RNA indicated that there is a single 1.7 kb GS transcript that increased during development to peak at the initial stages of culmination. Furthermore, GS mRNA was highly localized in prespore cells, which is consistent with a proposed source-sink model for ammonia assimilation in this organism. PMID- 9338598 TI - Involvement of the protein of Xenopus vasa homolog (Xenopus vasa-like gene 1, XVLG1) in the differentiation of primordial germ cells. AB - In order to understand the role of the protein of Xenopus vasa homolog (Xenopus vasa-like gene 1, XVLG1) in germ line cells, an attempt was made to perturb the function of the protein with the anti-vasa antibody 2L-13. The 2L-13 or the control antibody was microinjected with a lineage tracer (FITC-dextran-lysine, FDL) into single vegetal blastomeres containing the germ plasm of Xenopus 32-cell embryos, the descendants of which were destined to differentiate into a small number of primordial germ cells (PGC) and a large number of somatic cells, mostly of endodermal tissues at the tadpole stage. No significant effect of the injection of the antibodies on FDL-labeled, presumptive PGC (pPGC) was observed in embryos until stage 37/38. However, FDL-labeled PGC were not observed in almost all the 2L-13 antibody-injected tadpoles, although a similar number of labeled somatic cells were always present. As 2L-13 antibody specifically reacts with XVLG1 protein in the embryos by immunoblotting, the present results suggest that the antibody perturbed the function of XVLG1 protein in the pPGC, resulting in failure of PGC differentiation at the tadpole stage. PMID- 9338600 TI - Early specification of intestinal epithelium in the chicken embryo: a study on the localization and regulation of CdxA expression. AB - CdxA, a chicken homeobox-containing gene related to caudal in Drosophila, has been implicated in the regionalization of endoderm. It is reported here that, in the development of the chicken embryo, CdxA expression appears in the endoderm at day 1.5 of development as bilateral bands on either side of the splanchnopleure which later contribute to intestinal epithelium. The CdxA-expressing area extends medially and caudally as formation of the gut tube progresses. It is also shown that the rostral limit of CdxA expression demarcates the boundary between stomach and duodenum after day 3 of development. CdxA is not expressed in digestive tract appendages which open into the intestine, such as pancreas, liver and allantois. Early restriction of CdxA expression in intestinal lineage suggests that the intestinal specification involving CdxA expression commences before the gut tube is formed. The expression of CdxA in epithelial-mesenchymal tissue recombinants suggests that mesenchymal influence regulating CdxA expression plays an important role in confirming the boundary between the stomach and intestine. Chronological change in the spatial distribution of CdxA transcripts and the results of tissue recombination experiments, together with precise fate maps of early endoderm and splanchnic mesoderm, lead to a model of mechanisms by which intestinal specification is brought about. PMID- 9338599 TI - Proliferation, differentiation and morphogenesis of fetal rat glandular stomach transplanted under the kidney capsule of syngeneic hosts. AB - Undifferentiated glandular stomach tissue fragments from 16.5-day fetal rats were transplanted under the kidney capsule of syngeneic adult rats, and the proliferation, differentiation and morphogenesis of the transplanted tissues were investigated. Gastric epithelial cells began to invaginate 3-4 days after the transplantation and immature glands were formed after 1 week. During the period, there was a gradual increase in the expression of pepsinogen and cathepsin E, markers of cytodifferentiation of the stomach epithelia, both at protein and mRNA levels. Cathepsin E was weakly expressed in undifferentiated gastric epithelial cells at 16.5 days of gestation, and a higher level of the expression was observed in differentiated epithelia of the transplants. In contrast, the pepsinogen-producing cells first appeared around days 3-4 after transplantation and gradually increased in number to about 30% of the epithelial cells and became localized at the bottom of the gland. During the period of the experiment up to 1 month, the pepsinogen-producing cells were all positive for class III mucin and cathepsin E, indicating the immature character of these cells. In addition, no parietal cells were observed. When the tissue fragments were transplanted into adrenalectomized animals, the epithelial differentiation and morphogenesis was suppressed, but its proliferation was enhanced. The observed changes were reversed by hydrocortisone replacement. These results suggest that the development of the 16.5-day fetal stomach is regulated intrinsically to a certain extent by the genetic program of the cells involved and various gastric functions develop in the absence of luminal stimulation, stage-specific systemic hormonal change, neuronal regulation or other systemic influences, and that glucocorticoids modulate the developmental program of the fetal stomach tissues. PMID- 9338601 TI - Evidence for cell surface and internal phospholipase activity in ascidian eggs. AB - Upon fertilization, ascidian eggs release a cell surface glycosidase used in the block to polyspermy and undergo cortical contractions resulting from increased intracellular calcium levels. The glycosidase is released by fertilization, calcium ionophores or added phospholipase C (PLC) activity. The PLC inhibitor D609 blocks glycosidase release. Intact Ascidia ceratodes eggs cleave 4 methylumbelliferyl-phospho-choline when it is added to seawater. This yields highly fluorescent 4-methylumbelliferone. Authentic phospholipase C but not phospholipase D can cleave this substrate. Thus, the authors believe that cleavage of the substrate is specific for PLC activity. Eggs incubated in the fluorogenic substrate after having been washed and detergent extracted were not fluorescent. Therefore the substrate failed to enter intact cells. Glycosidase release and PLC activity were stimulated by ionomycin. Octylglucoside or Triton X 100 extracts of ascidian eggs had two forms of phospholipase activity as shown by ion affinity chromatography: PL1 eluting at 0.25 mol/L NaCl and PL2 eluting at 0.6 mol/L NaCl. The PL1 appeared to be isolated as a single protein. When surface proteins were labeled with non-penetrating biotin and were subsequently reacted with streptavidin, half of the PLC activity bound. This demonstrates that half the ascidian egg PLC activity is located on the surface of either the egg or follicle cell, and half is located within the egg. PMID- 9338602 TI - Immunological control of Trypanosoma cruzi infection and pathogenesis of Chagas' disease. AB - The major goal of studies on immunity to Trypanosoma cruzi is the understanding of immunological mechanisms involved in resistance to this protozoan as well as the pathogenesis of Chagas' disease. Different studies have defined CD8+ T lymphocytes, IFN-gamma and macrophages as important elements controlling parasite replication during the acute phase of infection. In contrast, during the chronic stage of the disease parasite-specific antibodies that fix complement and lyse the blood from trypomastigotes are thought to be the main effector molecules responsible for maintaining latent infection. In both acute and chronic infection with T. cruzi CD4+ Th1 lymphocytes appear to be the main cells responsible for induction of protective immunity. The immunological mechanisms involved in the pathogenesis of Chagas' disease are much more controversial. CD8+ lymphocytes are thought to be the main effector cells responsible for cardiac tissue destruction. Although many experiments suggest the involvement of autoimmunity in the pathogenesis of Chagas' disease, recent studies both in mice and humans indicate a positive association of tissue parasitism, inflammation and severity of pathology induced by T. cruzi. Finally, T. cruzi has emerged as an important opportunistic pathogen in patients infected with HIV and presenting low numbers of CD4+ T lymphocytes. These clinical findings indicate the essential requirement of CD4+ T-helper cells in mediating resistance during chronic Chagas' disease; however, the effector mechanisms that control the reactivation of chronic infection in vivo are not completely defined. PMID- 9338603 TI - Neutrophil-induced endothelial cell damage: inhibition by a 14-membered ring macrolide through the action of nitric oxide. AB - Macrolide antibiotics have been used worldwide for about 40 years. The clinical effectiveness of oral erythromycin for diffuse panbronchiolitis has been established and erythromycin seems to act not only as an antibacterial but also as an anti-inflammatory agent. We investigated the effect of 14-membered ring macrolides, erythromycin and clarithromycin, on human neutrophil functions and endothelial cell damage induced by neutrophils. The superoxide production of neutrophils and Ca2+ influx into neutrophils induced by N-formyl-methionyl-leucyl phenylalanine was inhibited by treatment with erythromycin but not by treatment with clarithromycin. When endothelial cells were cocultured with neutrophils, nitric oxide (NO) presumably released from endothelial cells were enhanced by treatment with erythromycin but not by treatment with clarithromycin and endothelial cell injury induced by neutrophils was ameliorated by addition of erythromycin but not by clarithromycin. The reduction of neutrophil-induced endothelial cell injury by erythromycin was abolished by treatment with carboxy PTIO which traps NO in the medium. Moreover, nitrite in the medium in which endothelial cells were incubated with neutrophils was enhanced by treatment with erythromycin and the enhancement of nitrite by erythromycin was partially cancelled by addition of H-89, an inhibitor of cyclic AMP-dependent protein kinase (PKA). Erythromycin seems to ameliorate neutrophil-induced endothelial cell injury by affecting not only neutrophil functions but the release of NO from endothelial cells through the action of PKA. The usefulness for the treatment of diseases worsened by the interaction between neutrophils and endothelium might be different among 14-membered ring macrolides. PMID- 9338604 TI - Immunoglobulin levels in patients with carbohydrate-deficient glycoprotein syndrome type I. AB - BACKGROUND: The characteristic feature of carbohydrate-deficient glycoprotein syndrome (CDGS) type I, a multisystemic disease, is underglycosylation of many serum glycoproteins, such as transferrin. A few cases of severe infections during childhood have been reported and an underlying immunodeficiency has been suggested. Because of this and the fact that all immunoglobulin (Ig) isotypes are glycoproteins we analysed the Ig levels in patients with CDGS I. METHODS: The serum concentrations of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgD and IgE, and the frequency of the G2m(23) allotype were measured by enzyme immunoassay in 15 patients with CDGS type I. RESULTS: Ten (67%) patients had an elevated level of at least one Ig, when compared to age-related reference ranges. No particular isotype was involved although a tendency towards high IgE levels was registered. The frequency of homozygous G2m(23)-negative CDGS patients (33%) was not different from that of blood donors (34%). CONCLUSION: We conclude that CDGS I patients have no major changes in the serum levels of any specific Ig isotype. The severe infections observed in some CDGS patients are therefore unlikely to involve any Ig deficiency. Our results do not exclude that Ig of patients with CDGS may have altered physiological functions because of abnormal glycosylation. PMID- 9338605 TI - Interleukin 7 and sCD23 synergize in the induction of human T cell activation in HIV-1-infected subjects. AB - The role played by CD23 in retroviral infections is still unclear. The synergistic effects of interleukin 7 (IL-7) and sCD23 on T cell proliferation and the generation of HIV-1-specific cytotoxic T lymphocytes (CTL) in mitogen- and antigen-stimulated systems were examined. Addition of IL-7 and sCD23 at the onset of culture resulted in a marked augmentation of T cell proliferation and cytotoxic activity. Studies of CTL development in purified mitogen CD8+ T cells demonstrated that IL-7 and sCD23 could act directly on the CD8+ lymphocyte subset to augment cytotoxicity. The data demonstrate that IL-7 and sCD23 synergistically augmented CTL activity independently of IL-2 and IL-12. We analyzed the effects on IFN-gamma production by CD8+ T cells and found that IL-7 alone did not induce detectable levels of IFN-gamma production, but together with sCD23, it synergistically enhanced the production of IFN-gamma. We also found that IFN gamma appeared not to be required for the enhanced CD8+ CTL activity because rabbit anti-IFN-gamma antibody did not block the synergistic effects of IL-7 and sCD23. These results indicate that IL-7 and sCD23 can exert major upregulatory effects on human CTL development and suggest that these effects are both proliferative and differentiative. PMID- 9338606 TI - Localization of granule proteins in human eosinophil bone marrow progenitors. AB - Eosinophils have a characteristic content of cationic proteins, stored in core containing specific granules and released at sites of inflammation; coreless granules (sometimes called primary) are present in eosinophil promyelocytes. In order to determine a possible relationship between the two granule subsets, immunoelectron-microscopic techniques were used to determine the presence and precise intragranular distribution of major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil peroxidase (EPO), and arylsulfatase B of eosinophil granules, as well as the Charcot-Leyden crystal (CLC) protein, in eosinophil progenitors of the bone marrow. MBP, ECP, EPO, and arylsulfatase B were observed in both coreless and core-containing (specific) granules. The difference in the distribution of MBP, having a uniform distribution in coreless granules and a crystalloid distribution in core-containing (specific) granules, could indicate a maturational process of a common organelle. CLC protein was distributed in the cytosol, in the euchromatin of the nuclei, but was also present in a rare granular compartment of both immature and mature eosinophils. The present findings suggest that coreless granules develop into core-containing specific granules. PMID- 9338607 TI - Secretory response of mast cells contained in monodispersed human choroidal preparations. AB - BACKGROUND: Mast cells have been identified in the choroid of numerous species including man. However, functional studies involving these human mast cells have not been reported. In the current studies, the secretory response of human choroidal mast cells to various stimuli was examined using monodispersed choroidal cell preparations. METHODS: Monodispersed cell suspensions of human choroid were prepared from eye bank globes and the number, histamine content, and secretory response of mast cells in these preparations were determined. Choroids from 27 donors were used for these experiments. RESULTS: Cell suspensions contained an average of 15% mast cells. Mast cells stained positively with toluidine blue and exhibited the classical granular appearance upon electron microscopy. The amount of histamine contained in each mast cell was calculated to be 2.74+/-0.17 pg. Significant histamine release was observed following treatment with anti-human IgE, calcium ionophore A23187, concanavalin A, compound 48/80 and morphine. CONCLUSION: A method has been developed for obtaining monodispersed human choroidal mast cell preparations. The cells were functional as evidenced by their ability to release histamine upon immunological and nonimmunological stimulation. The degranulation noted following compound 48/80 and morphine challenge suggests that these human choroidal mast cells are analogous to connective tissue or chymase/tryptase-positive mast cells. PMID- 9338608 TI - Diagnosis and carrier detection of chronic granulomatous disease in five families by flow cytometry. AB - BACKGROUND: The application of flow cytometric assays, for determination of phagocyte respiratory burst (ROB) activity, to the investigation of chronic granulomatous disease (CGD) may lead to improved laboratory detection of patients and carriers and indicate the nature of the molecular defect. To evaluate the diagnostic capability of flow cytometry an investigation of 5 CGD families was undertaken. METHODS: Phorbol myristate acetate (PMA)-induced neutrophil ROB was determined using dihydrorhodamine 123 (DHR) and flow cytometric analysis in 26 members of 5 CGD families (2: X-CGD; 3: autosomal recessive CGD). RESULTS: Neutrophils from X-CGD patients displayed absence of reactivity. Female carriers demonstrated dual fluorescence peaks of high and low intensity indicative of normal and abnormal populations, respectively. Normal ROB activity was observed in a boy whose X-CGD was successfully treated by bone marrow transplantion. Reduced ROB activity was observed in 3 patients with autosomal-recessive CGD compared with their parents and siblings. The patterns of flow cytometric reactivity correlated with the different molecular defects identified. Absence of the p22phox membrane component of the NADPH oxidase complex resulted in a significantly reduced level of respiratory burst activity which was comparable to that observed in X-CGD, whereas reduced but detectable levels of respiratory burst activity were observed in a patient with diminished levels of p22phox and in a patient with deficiency of the cytosolic p47phox component. CONCLUSIONS: The DHR flow cytometric assay offers a sensitive diagnostic screening test for CGD and furthermore may provide an indication of the likely underlying molecular defect. PMID- 9338609 TI - Decreased production of interleukin-1-beta, prostaglandin-E2 and thromboxane-B2, and elevated levels of interleukin-6 and -10 are associated with increased survival during endotoxic shock in mice consuming diets enriched with sesame seed oil supplemented with Quil-A saponin. AB - Sesamin, present in sesame seed oil (SSO), can inhibit delta-5-desaturase activity and cause accumulation of dihomo-gamma-linolenic acid (DGLA), which displaces arachidonic acid, and subsequently decrease production of dienoic eicosanoids. The effects of diets containing both SSO and Quil A, a saponin that emulsifies fats and potentiates the immune responses, were also studied. A mixture of oils having a fatty-acid composition similar to that of SSO served as a control diet. The levels of docosapentaenoic acid in mice fed Quil-A supplemented diets and of DGLA in those fed SSO diets were markedly higher in the liver. These changes were associated with a significant reduction in the plasma prostaglandin-E(1+2) and thromboxane-B2 levels in response to an intraperitoneal injection of a lethal dose of lipopolysaccharide (LPS) endotoxin (LD50 20 mg/kg). The levels of interleukin (IL-)6 were elevated and those of IL-1beta were decreased in mice consuming Quil-A-supplemented diets. The IL-10 levels that were elevated in all mice after LPS exposure, remained higher (even at 9 h) only in those fed Quil-A-supplemented diets, but declined rapidly in others. During a 48 hour observation period following LPS injection, all control animals died, and survival was 40% in the SSO group, and 27 and 50%, respectively, in those fed Quil-A-supplemented control and SSO diets. These data suggest that SSO and Quil A when present in the diet exerted cumulative effects that resulted in a decrease in the levels of dienoic eicosanoids with a reduction in IL-1beta and a concomitant elevation in the levels of IL-10 that were associated with a marked increase in survival in mice. PMID- 9338610 TI - Anti-Thy-1 antibody provokes reverse passive cutaneous anaphylaxis-like reactions. AB - Antibodies to rat Thy-1, and also to murine Thy-1, could provoke reverse passive cutaneous anaphylaxis-like (RPCA-like) reactions. The threshold amount of purified anti-Thy-1 antibodies for RPCA-like reactions was 12.5 microg/ml. No RPCA-like reactions were obtained with IgG1 F(ab')2 anti-Thy-1. Cross-linking the murine anti-rat Thy-1 F(ab')2 fragments with goat antibody against murine F(ab')2 anti-rat Thy-1 permitted us to obtain RPCA-like reactions with as little as 1.5 microg/ ml. However, when the cross-linking was done with the F(ab')2 of the same goat antibody, no RPCA-like reactions were obtained. The antibody against rat FcepsilonRIalpha or its F(ab')2 product were equally effective in producing RPCA like reactions; the threshold was obtained with 1.5 microg/ml. The mechanism of the difference is discussed. PMID- 9338611 TI - Diesel exhaust particulates enhance eosinophil adhesion to nasal epithelial cells and cause degranulation. AB - Diesel exhaust particulates (DEP) are a common air pollutant from diesel-engine powered car exhaust and are thought to cause chronic airway diseases. On the other hand, eosinophils are major components of allergic inflammatory disorders such as asthma, nasal allergy and atopic dermatitis. We examined the effects of DEP and DEP extract (extract of polyaromatic hydrocarbons) on eosinophil adhesion, survival rate and degranulation. Eosinophils, human mucosal microvascular endothelial cells (HMMECs) and human nasal epithelial cells (HNECs) were preincubated in the presence or absence of DEP and DEP extract. 35S-labeled eosinophils were allowed to adhere to monolayers of HMMECs and HNECs. After washing, 35S radioactivity was determined and numbers of adherent eosinophils were calculated using each standard curve. The effects of DEP and DEP extract on eosinophil survival rate and degranulation were also determined. Although neither DEP nor DEP extract affected the adhesiveness of HMMECs and HNECs to eosinophils, 5 ng/ml of DEP extract and 50 ng/ml of DEP extract each significancy increased eosinophil adhesiveness to HNECs (134+/-9 and 143+/-8%, respectively; p<0.01 vs. control), but neither effected eosinophil adhesiveness to HMMECs. DEP extract also induced eosinophil degranulation without changing the eosinophil survival rate. Given that eosinophil-derived lipid mediators and toxic proteins play important roles in the development of nasal allergy, the above findings strongly suggest that DEP plays an important role in promoting the nasal hypersensitivity induced by enhanced eosinophil infiltration of epithelium and eosinophil degranulation. PMID- 9338612 TI - Maternal influence on IgG subclass antibodies to Bet v 1 during the first 18 months of life as detected with a sensitive ELISA. AB - BACKGROUND: The initial encounters with allergens are crucial for sensitisation later in life. The IgG1 responses to house dust mite in infancy are later accompanied by an IgG4 response, with high levels seen particularly in atopic children. Little is known of the development of IgG subclass responses to other inhalant allergens. The aims of this study were to develop a sensitive method for the study of postnatal immune responses to the important seasonal inhalant allergen Bet v 1. METHODS: Antibodies to rBet v 1 were analyzed in 96 children at birth, 6 and 18 months using a sensitive enzyme-amplified ELISA. RESULTS: Immunoglobulin G responses to rBet v 1, mainly of the IgG1 subclass, were common in young children, and could at 6 months be demonstrated in several infants who had not yet experienced a postnatal birch pollen season. Atopic dermatitis was associated with high levels of IgG subclass antibodies to birch at 18 months. Maternal atopy was associated with high levels of all IgG subclass antibodies to rBet v 1 in cord blood. In postnatally birch-pollen-exposed infants with atopic mothers, the levels of IgG antibodies at birth correlated with the levels at 6 months. In contrast, high antibody levels to rBet v 1 at birth were associated with low IgG titres to rBet v 1 at 18 months. CONCLUSIONS: IgG1 responses to birch are common during the first 18 months of life. High levels of maternally derived birch-specific IgG antibodies are associated with maternal atopy and may influence the development of the IgG antibody responses to birch in their child. PMID- 9338613 TI - Effect of topical applications of budesonide and azelastine on nasal symptoms, eosinophil count and mediator release in atopic patients after nasal allergen challenge during the pollen season. AB - We studied the activity of a topical form of a corticosteroid (budesonide) and an antihistamine (azelastine) in the treatment of seasonal allergic rhinitis by including an assessment of mediator concentrations and the percentage of eosinophils in the nasal secretions before and after the treatment. Nasal allergen challenge (NAC) during the season was performed to mimic an acute attack of allergic rhinitis and to objectively evaluate the effect of the drugs on the early-phase reaction. The study compared in a randomized way (2 parallel groups) the effect of budesonide (Rhinocort Aqua) and azelastine (Allergodil nasal spray) in a group of 14 patients during the pollen season. The study showed that azelastine significantly reduced sneezing, total nasal resistance and increased nasal airflow even when significant increases in histamine, tryptase and leukotriene C4 (LTC4) concentrations in nasal secretions were evidenced immediately after NAC. Budesonide showed a strong (p<0.05) decrease in infiltration and activation of eosinophils, and on tryptase and LTC4 release after NAC. These effects (not for LTC4) lasted at least for 1 week after therapy. Azelastine is a powerful topical antihistamine, while budesonide appears to be a potent long-acting anti-inflammatory agent. PMID- 9338614 TI - Relationship of upper airway disease to tobacco smoking and allergic markers: a cohort study of men followed up for 5 years. AB - Data derived from a cohort study of 191 men, seen 5 years apart, were used to investigate the involvement in allergic as well as in nonallergic upper airway disease (UAD) of two risk factors, immediate hypersensitivity and tobacco smoking, the roles of which have been well established in lower airway disease. At both surveys, UAD and smoking habits were assessed by an extended version of the BMRC/ECSC questionnaire. UAD consisted of usual or chronic rhinitis, seasonal allergic rhinitis and perceived nasal hyperresponsiveness (PNHR) to tobacco smoke, cold air or exercise. Immediate hypersensitivity was determined either in vivo (skin prick test, SPT, positivity) or in vitro (total IgE). UAD prevalence and smoking habits did not vary significantly over 5 years. On the contrary, SPT positivity increased significantly between the two surveys. At both surveys, SPT positivity for common aeroallergens (grass pollens overall) was significantly related to seasonal allergic rhinitis but not to usual or chronic rhinitis and to PNHR. Similarly, the total IgE level was increased in seasonal allergic rhinitis, but never significantly. Current smoking was always a habit significantly more frequent in men reporting chronic rhinitis (odds ratios of 5.3 and 4.9 in 1985 and 1990, respectively). The relationship was of the dose-response type: the more the subjects smoked, the more they reported chronic rhinitis. In contrast, seasonal allergic rhinitis was more associated with exsmoking, either in 1985 or in 1990. These results were confirmed longitudinally and after exclusion of asthmatics. Further investigations are needed to support the hypothesis raised by our data according to which immediate hypersensitivity, as assessed by SPT positivity for common aeroallergens, and tobacco smoking might intervene alternatively in UAD, probably because of the 'healthy smoker effect'. PMID- 9338615 TI - Lack of lysozyme activity of natural and yeast-derived recombinant Der p 2. AB - For Dermatophagoides pteronyssinus a number of allergens have been reported. Although the function of several allergens is known, that of the house dust mite allergen Der p 2 is unknown. A role as lysozyme has been suggested. We tested a yeast-derived recombinant Der p 2 for its lysozymatic activity. This recombinant allergen and affinity-purified natural Der p 2 lacked this enzymatic activity. Whole-body mite extract retained its lysozymatic activity after removal of the group 2 allergen by monoclonal antibody depletion. Analysis of spent medium revealed the reciprocal: it contained group 2 allergen, but lacked lysozymatic activity. Together, these data demonstrate that mite lysozyme and Der p 2 are different components of mite extract. PMID- 9338616 TI - Hypersensitivity pneumonitis related to a covered and heated swimming pool environment. AB - Hypersensitivity pneumonitis (HP) or extrinsic allergic alveolitis is a lung disease caused by a large group of inhaled antigens of various sources. The most common HP occurring in the farm environment is classically caused by exposure to various thermophilic actinomycetes and fungi that can grow in the farm environment. Pullularia species and thermophilic actinomycetes have been involved in HP related to humidifier water and saunas. Our case illustrates the value of a site visit in the diagnosis of HP. During a visit to the covered and heated swimming-pool where our patient used to swim we could see that favourable conditions to fungal growth existed. To determine the possible aetiological agents of a suspected HP, cultures from several parts of the swimming-pool were taken. These cultures showed an intense growth of thermophilic actinomycetes, Neurospora and Aspergillus species. Precipitating antibodies against Neurospora species and Mycropolyspora faeni were detected. A case of HP related to a covered and heated swimming-pool environment is reported. Thermophilic actinomycetes and Neurospora species may be the causing agents. PMID- 9338617 TI - Acquired pure red cell aplasia in Japan. AB - We reviewed the clinical features of 150 patients with acquired pure red cell aplasia (PRCA) in Japan. There were 35 patients with acute type and 115 with chronic type PRCA. Of the acute PRCA patients, 17 had human parvovirus B19 infection. Drug-induced PRCA was demonstrated in 7 patients. Of the 115 patients with chronic PRCA, 51 patients were classified as primary and 64 cases were associated with miscellaneous diseases such as thymoma, a variety of hematological disorders and collagen diseases. Among the hematological disorders, PRCA was most frequently seen in granular lymphocyte proliferative disorders (GLPD). The erythroid colony growth patterns from bone marrow were variable. The serum erythropoietin level was high in most patients. Various kinds of treatment were tried for the chronic PRCA cases. Cyclosporin A (CyA) was the most effective form of treatment and the response rate was 82% (31/38). Twenty-three of 37 patients (62%) responded to bolus methylprednisolone therapy. The largest number of patients were treated with oral prednisolone, and the therapy was effective in 27 of the 55 (49%). The response rate to cyclophosphamide was only 29% (5/17), but in combination with prednisolone, half of the patients (7/14) responded to the therapy. CyA is recommended as the first-line therapy for acquired chronic PRCA. PMID- 9338618 TI - Five putative drug resistance parameters (MDR1/P-glycoprotein, MDR-associated protein, glutathione-S-transferase, bcl-2 and topoisomerase IIalpha) in 57 newly diagnosed acute myeloid leukaemias. Swiss Group for Clinical Cancer Research (SAKK). AB - Using a modified quantitative reverse transcriptase (RT) PCR assay in 57 patients with acute myeloid leukaemia (AML) from a Swiss Phase III multicentre study (SAKK 30/85), we measured the m-RNA expression of the genes from the multidrug resistance gene 1 (MDR1), the multidrug resistance associated protein (MRP), glutathione-S-transferase (GST) pi, bcl-2 and topoisomerase (topo) IIalpha. P glycoprotein (p-gp) was measured by Western blot, and GST activity by functional assays. To analyse progression-free (PFS) and overall survival (OS), parameters were prospectively divided into "low" and "high" groups, according to their median values (exceptions: MDR1 and p-gp). Median follow-up was 60 months. RESULTS: MDR1- and MRP mRNA levels correlated with each other (r=0.54, Spearman), FABM4/M5 and extramedullary disease. "Low" bcl-2-mRNA predicted longer PFS: 22 months vs. 7 months (median,p=0.02, log rank), and longer OS: 64 months vs. 14 months (p=0.06). "Low" topo IIalpha predicted poorer outcome: median PFS 9 vs. 19 months (p=0.03); median survival 12 months vs. "not reached" (p=0.03). An improved outcome tendency, albeit nonsignificant, was seen in p-gp-negative patients. In a Cox model adjusted for age, performance status, presence of Auer rods, FAB type and clinical response, bcl-2 and topo IIalpha mRNA levels retained their predictive values. PMID- 9338619 TI - 2-Chlorodeoxyadenosine (cladribine)-related eosinophilia in patients with lymphoproliferative diseases. AB - Eosinophilia and allergic skin reactions are uncommon events after 2 chlorodoxyadenosine (2-CdA, cladribine) administration. A multicentre retrospective analysis of eosinophilia in 360 patients treated with 2-CdA for lymphoid malignancies has been made. B-cell chronic lymphocytic leukaemia (B-CLL) was diagnosed in 153, hairy cell leukaemia (HCL) in 68, low-grade non-Hodgkin's lymphoma (LGNHL) in 119, high-grade NHL in 2 and Waldenstrom's macroglobulinaemia (WM) in 18 patients. 2-CdA was administered at a dose 0.12 mg/kg/d in 2-h intravenous infusion for 5 consecutive d. The courses were repeated monthly. Patients with HCL received 1 cycle of 2-CdA, with NHL 2-6 (mean 3.5) cycles and with B-CLL 3-6 (mean 5) cycles. Twenty patients (5.5%), including 5 with HCL, 6 with LGNHL, 7 with B-CLL and 2 with WM, developed peripheral blood eosinophilia. The mean values of absolute eosinophil count were 0.78x10(9)/l (0.58 1.06x10(9)/l), 0.71x10(9)/l (0.52-1.3x10(9)/l), 85 (0.56-1.82x10(9)/l) and 0.75 (0.74-0.76x10(9)/l), respectively. Eosinophilia occurred in 13 patients after 1 course, in 4 after 2 courses, and in 5 after > or =3 courses of the therapy. In 17 cases it resolved spontaneously. Allergic skin lesions with pruritus were noticed in 3 patients simultaneously with an increase in eosinophil count. All of them required antihistaminic drugs and/or corticosteroids. One patient with B-CLL experienced repeated episodes of eosinophilia. The highest incidence of 2-CdA induced eosinophilia was noticed in patients with MW (11.1%) and HCL (7.4%) who received only 1 cycle of this drug and entered a complete remission. This side effect was less frequently observed in LGNHL and B-CLL, i.e. in 5.0% and 4.6% of cases, respectively. The mechanism of 2-CdA-induced eosinophilia is not clear. It has been postulated that massive tumour cell lysis may trigger a release of IL-5 and probably other cytokines. The allergic mechanism of 2-CdA-induced eosinophilia is also possible, especially in patients with simultaneous skin reactions. PMID- 9338621 TI - Distribution and incidence rates of lymphoid neoplasms according to the REAL classification in a single institution. A prospective study of 940 cases. AB - Recently, a new classification system for lymphoid neoplasms, known as the REAL classification, has been proposed. Our aim is to know the distribution of lymphoid neoplasms according to this schema and compare it with the Updated Kiel classification. We also estimate incidence rates of lymphoid neoplasms in our area. From January 1993 to November 1996, 940 patients were diagnosed of lymphoid neoplasm in our center. Histologic material was prospectively classified according to both the REAL and the Updated Kiel classifications. According to the REAL classification, distribution of all cases of lymphoid neoplasms was as follows: 73.6% B-cell neoplasm, 9.4% T-cell neoplasms, 9.6% Hodgkin's disease and 7.4% unclassifiable. Considering only non-Hodgkin's lymphomas (NHL), 87.2% of cases could be categorized according to the REAL and 77.7% with the Updated Kiel classification. These figures differed due to unrecognized categories in the Kiel schema. Annual incidence rate per 100,000 inhabitants was 20.1 for lymphoid neoplasms, and NHL alone was 9.0. In conclusion, the REAL classification allowed us to categorize more cases of NHL than did the Updated Kiel classification, fundamentally because of the inclusion of some recently described entities. PMID- 9338620 TI - Cardiac function during iron chelation therapy in adult non-thalassaemic patients with transfusional iron overload. AB - It is well-documented that iron chelation by desferrioxamine protects/improves the cardiac function in blood transfusion-dependent children suffering from beta thalassaemia. In patients who do not become dependent upon blood transfusion until adulthood (ANT-patients), iron chelation by desferrioxamine may affect the cardiac function in unknown ways, presumably because age-related changes in the heart may cause iron chelation to affect the cardiac function in different ways. We therefore followed the left ventricular ejection fraction (LVEF) by multigated radionuclide angiography in 16 iron-loaded ANT-patients during iron chelation alone and after increasing the efficacy of chelation by vitamin C supplementation. During 12 months of iron chelation the mean LVEF fell significantly from 63.3% to 58.0% (p=0.04). Individual changes in LVEF did not correlate significantly with age but with the pretreatment liver iron concentration. After initiation of vitamin C supplementation, the mean LVEF increased from 55.9% to 65.3% (p=0.01). Our data suggest that in ANT-patients prolonged desferrioxamine treatment without vitamin C supplementation may be associated with reduced LVEF, whereas vitamin C supplementation seems to benefit the cardiac function. Similar findings have not been described in beta thalassaemia and may hence be specific for ANT-patients. However, our findings have to be confirmed by controlled studies. PMID- 9338622 TI - Production of recombinant human GM-CSF-EPO hybrid proteins: in vitro biological characterization. AB - Selective lineage differentiation depends upon the combined action of several colony-stimulating factors. Here we describe 3 human granulocyte-macrophage colony-stimulating factor-erythropoietin (GM-CSF-EPO) hybrid proteins generated by recombination of the relevant cDNAs. The expression vector containing the murine cytomegalovirus (mCMV) promoter and dihydrofolate reductase (DHFR) gene was used for the expression of the hybrid genes in Chinese hamster ovary (CHO) cells. Purified hybrid proteins from CHO transfectant cultures induced proliferation of both EPO and GM-CSF dependent cell lines. The clonogenic test, performed on purified human hematopoietic precursor cells, indicates that the hybrid proteins are more efficient at inducing erythroid differentiation compared with the equimolar mixture of GM-CSF and EPO. PMID- 9338623 TI - Prognostic factors of hemophagocytic syndrome in adults: analysis of 34 cases. AB - Hemophagocytic syndrome (HPS) presents with fever, pancytopenia, liver dysfunction and increase in hemophagocytic histiocytes in various organs. Although there are two major classifications of HPS in adults, malignant and reactive histiocytosis, it is often very difficult to distinguish between these disorders. We analyzed the laboratory data of patients with HPS to evaluate prognostic factors. Of 34 patients, 14 survived, and 20 died. The median age of survivors was 29.6+/-11.5 yr significantly younger than those who died (54.7+/ 17.8 yr). Twenty patients had no obvious underlying disease, the other 13 had hematological malignancies or viral infections. Comparison of laboratory data revealed that nonsurvivors had significantly lower Hb and platelet values on admission. During treatment, worsening of anemia and thrombocytopenia, increase of transaminase and biliary enzymes were similarly more prominent. Risk factors associated with death were: age over 30 yr, presence of disseminated intravascular coagulation, increased ferritin and beta2-microglobulin, anemia accompanied by thrombocytopenia and jaundice. Our data suggests that patients with HPS and any of these risk factors should be treated aggressively with sufficient chemotherapy and supportive care. PMID- 9338624 TI - Serum levels of cytokines after bone marrow transplantation: increased IL-8 levels during severe veno-occlusive disease of the liver. AB - We investigated the cytokine profile and peak levels of interleukin (IL) -6, IL 8, IL-10 and tumour necrosis factor (TNF) -alpha levels in 42 patients after allogeneic bone marrow transplantation (BMT). Eleven of them developed veno occlusive disease (VOD) of the liver. Fourteen patients had moderate-to-severe acute graft-versus-host disease (aGvHD), 10 isolated bacteraemia and 7 had no major complication. Those who developed severe VOD (n=6) showed a short, very high IL-8 peak (median: 6632 pg/ml, range: 5546-10,000 vs. 280 pg/ml, 0-2042 in controls, p<0.01) 1-4 d after diagnosis of the liver disease. Five of these patients had high peak levels of IL-6. Five patients with mild VOD showed a lower increase in the cytokines tested. Bilirubin levels, at day of IL-8 peak, did not differ statistically between mild and severe VOD. The highest levels of IL-10 were found in those with aGvHD. IL-8 levels were also increased, but not to the same extent as in patients with severe VOD (p=0.01 vs. VOD). In patients with bacteraemia, very high levels of IL-6 were seen. In patients without major complications, the levels of cytokines were low. In conclusion, high levels of IL 8 occurred in severe VOD of the liver, which may be of value to determine prognosis. PMID- 9338625 TI - Recombinant human erythropoietin for the treatment of anemia in chronic myelogenous leukemia. PMID- 9338626 TI - Concomitant in vitro platelet hypofunction and increased thromboxane B2 (TXB2) generation and enhanced in vivo platelet activation. A distinct syndrome in thrombosis? PMID- 9338627 TI - Intramembrane protein distribution in cell cultures is affected by 50 Hz pulsed magnetic fields. AB - Intramembrane proteins (IMP) represent a class of proteins located in the lipid bilayer of the cell membrane which function as ion channels, enzymes or receptors. Since it has been argued that biological effects of extremely low frequency (ELF) electromagnetic fields are mediated by plasma membrane. this work was designed to study the possible effects of 50 Hz pulsed magnetic fields (PMF) of the type used to stimulate bone repair, on the distribution of IMP in the plasma membrane of Swiss NIH 3T3 fibroblasts. Evaluations were based on the calculation of a distribution factor, which allows discrimination between random, regular and clustered distribution of IMP, in electron microscope images of freeze-fractured membranes. The results indicate that cells exposed to PMF for more than two hours have a significant clustering of the IMP distribution compared to control unexposed cells. PMID- 9338628 TI - Exposure of welders and other metal workers to ELF magnetic fields. AB - This study assessed exposure to extremely low frequency (ELF) magnetic fields of welders and other metal workers and compared exposure from different welding processes. Exposure to ELF magnetic fields was measured for 50 workers selected from a nationwide cohort of metal workers and 15 nonrandomly selected full-time welders in a shipyard. The measurements were carried out with personal exposure meters during 3 days of work for the metal workers and I day of work for the shipyard welders. To record a large dynamic range of ELF magnetic field values, the measurements were carried out with "high/low" pairs of personal exposure meters. Additional measurements of static magnetic fields at fixed positions close to welding installations were done with a Hall-effect fluxmeter. The total time of measurement was 1273 hours. The metal workers reported welding activity for 5.8% of the time, and the median of the work-period mean exposure to ELF magnetic fields was 0.18 microT. DC metal inert or active gas welding (MIG/MAG) was used 80% of the time for welding, and AC manual metal arc welding (MMA) was used 10% of the time. The shipyard welders reported welding activity for 56% of the time, and the median and maximum of the workday mean exposure to ELF magnetic fields was 4.70 and 27.5 microT, respectively. For full-shift welders the average workday mean was 21.2 microT for MMA welders and 2.3 microT for MIG/MAG welders. The average exposure during the effective time of welding was estimated to be 65 microT for the MMA welding process and 7 microT for the MIG/MAG welding process. The time of exposure above 1 microT was found to be a useful measure of the effective time of welding. Large differences in exposure to ELF magnetic fields were found between different groups of welders, depending on the welding process and effective time of welding. MMA (AC) welding caused roughly 10 times higher exposure to ELF magnetic fields compared with MIG/MAG (DC) welding. The measurements of static fields suggest that the combined exposure to static and ELF fields of MIG/MAG (DC) welders and the exposure to ELF fields of MMA (AC) welders are roughly of the same level. PMID- 9338629 TI - Influence of human model resolution on computed currents induced in organs by 60 Hz magnetic fields. AB - The effects of human body model resolution on computed electric fields induced by 60 Hz uniform magnetic fields are investigated. A recently-developed scalar potential finite difference code for low-frequency electromagnetic computations is used to model induction in two anatomically realistic human body models. The first model consists of 204290 cubic voxels with 7.2-mm edges, while the second comprises 1639146 cubic voxels with 3.6-mm edges. Calculations on the lower resolution model using, for example, the finite difference time domain or impedance methods, push the capabilities of workstations. The scalar method, in contrast, can handle the higher-resolution model using comparable resources. The results are given in terms of average and maximum electric field intensities and current density magnitudes in selected tissues and organs. Although the lower resolution model provides generally acceptable results, there are important differences that make the added computational burden of the higher-resolution calculations worthwhile. In particular, the higher-resolution modelling generally predicts peak electric fields intensities and current density magnitudes that are slightly higher than those computed using the lower-resolution modelling. The differences can be quite large for small organs such as glands. PMID- 9338630 TI - Influence of extremely low frequency electromagnetic fields on the swimming behavior of ciliates. AB - Different species of ciliates (Paramecium biaurelia, Loxodes striatus, Tetrahymena thermophila) have been taken as model systems to study the effects of extremely low-frequency electromagnetic fields (50 Hz, 0.5-2.0 mT) on the cellular level. A dose-dependent increase in the mean swimming velocity and a decrease in the linearity of cell tracks were observed in all wild-type cells. In contrast, field-exposure did not increase the number of directional turns of the Paramecium tetraurelia pawn mutant (d4-500r), which is characterized by defective Ca2+-channels. The described changes indicate a direct effect of low frequency electromagnetic fields on the transport mechanisms of the cell membrane for ions controlling the motile activity of cilia. PMID- 9338631 TI - Stress proteins are not induced in mammalian cells exposed to radiofrequency or microwave radiation. AB - The induction of stress proteins in HeLa and CHO cells was investigated following a 2 h exposure to radiofrequency (RF) or microwave radiation. Cells were exposed or sham exposed in vitro under isothermal (37 +/- 0.2 degrees C) conditions. HeLa cells were exposed to 27- or 2450 MHz continuous wave (CW) radiation at a specific absorption rate (SAR) of 25 W/kg. CHO cells were exposed to CW 27 MHz radiation at a SAR of 100 W/kg. Parallel positive control studies included 2 h exposure of HeLa or CHO cells to 40 degrees C or to 45 microM cadmium sulfate. Stress protein induction was assayed 24 h after treatment by electrophoresis of whole-cell extracted protein labeled with [35S]-methionine. Both cell types exhibited well-characterized responses to the positive control stresses. Under these exposure conditions, neither microwave nor RF radiation had a detectable effect on stress protein induction as determined by either comparison of RF exposed cells with sham-exposed cells or comparison with heat-stressed or Cd++ positive control cells. PMID- 9338633 TI - Biological effects of continuous exposure of embryos and young chickens to electromagnetic fields emitted by video display units. AB - The effects of continuous exposure of embryos and young chickens to electromagnetic fields (EMFs) emitted by video display units (VDUs) were investigated. Embryos and brood were continuously exposed during embryonic and postembryonic phases to EMFs emitted by two types of VDU (TV or computer). Embryonic mortality was evaluated in three independent experiments. Young chickens were immunized three times by porcine thyroglobulin (Tg). Blood samples were assayed after each immunization for specific anti-Tg antibodies (IgG), plasma corticosterone (CORT), and plasma melatonin (MLT). In the sham-exposed samples, embryonic death (10-33%) was restricted to the perinatal period and the IgG, CORT, and MLT responses of young chickens crested after the second immunization. Constant EMF exposure was accompanied by significantly increased fetal loss (47-68%) and markedly depressed levels of circulating anti-Tg IgG, CORT, and MLT. Collectively, these findings indicate that continuous exposure to EMFs, issuing from VDUs, adversely affects embryos and young chickens. PMID- 9338632 TI - Magnetic field effects on coenzyme B12-dependent enzymes: validation of ethanolamine ammonia lyase results and extension to human methylmalonyl CoA mutase. AB - Enzymes with radical-pair intermediates have been considered as a likely target for purported magnetic field effects in humans. The bacterial enzyme ethanolamine ammonia lyase and the human enzyme methylmalonyl-CoA mutase catalyze coenzyme B12 dependent rearrangement reactions. A common step in the mechanism of these two enzymes is postulated to be homolysis of the cobalt-carbon bond of the cofactor to generate a spin-correlated radical pair consisting of the 5'-deoxyadenosyl radical and cob(II)alamin [Ado. Cbl(II)]. Thus, the reactions catalyzed by these enzymes are expected to be sensitive to an applied magnetic field according to the same principles that control radical pair chemical reactions. The magnetic field effect on ethanolamine ammonia lyase reported previously has been corroborated independently in one of the authors' laboratory. However, neither the human nor the bacterial mutase from Propionibacterium shermanii exhibits a magnetic field effect that could be greater than about 15%, considering the error limit imposed by the uncertainty of the coupled assay. Our studies suggest that putative magnetic field effects on physiological processes are not likely to be mediated by methylmalonyl-CoA mutase. PMID- 9338634 TI - Vertical circularly polarized ELF magnetic fields and induced electric fields in culture media. AB - Some properties of induced electric fields in cell culture media produced by vertical circularly polarized magnetic fields are examined. The described geometry is not advantageous for determining effects that may be attributable to induced electric fields or currents. PMID- 9338635 TI - Comments on "Resonance effect of millimeter waves in the power range from 10(-19) to 3 X 10(-3) W/cm2 on Escherichia coli cells at different concentrations," Belyaev et al., Bioelectromagnetics, 17:312-321 (1996) PMID- 9338636 TI - Managed care and medical ethics. PMID- 9338637 TI - Clinical and laboratory evaluation of a new thin-walled self-expanding tracheobronchial silicone stent: progress and pitfalls. AB - BACKGROUND: Although widely established in the management of malignant airway lesions, currently available tracheobronchial prostheses made of silicone have their drawbacks resulting from rigidity and wall thickness. Therefore we present clinical follow-up data obtained with a novel thin-walled expandable prototype silicone airway stent (Polyflex stent, Willy Rusch AG, Kernen, Germany) in 19 patients. METHODS: Seventeen of 19 patients had tracheobronchial complications of infiltrating cancer: five had respiratory-digestive fistulas, 14 had mixed-type obstructions (mucosal infiltration plus extrinsic compression), and two had diffuse tracheal hemorrhages from the tumor surface (three patients had more than one complication). Two of 19 patients had benign postintubation stricture and malacia. Overall, 33 stents were implanted either simultaneously or in a consecutive manner. Scanning electron microscopy was performed both on prototype stents and on other available silicone stents for comparison. RESULTS: The treatment improved the patients' clinical condition substantially. The mechanical properties of the new prosthesis were excellent. Important stent-associated side effects were early mucus retention (n = 7), infolding of the inner silicone layer (n = 2), and stent dislodgment (n = 2). As of February 1997, 10 patients have died of causes unrelated to stent placement. Seven patients with malignant airway disease are still alive from 2 weeks up to 7 months after initial treatment. Scanning electron microscopy of explanted and unused prototypes suggested that an extremely ragged luminal microstructure may contribute to the firm adhesion of secretory material and that technical smoothing of the surface avoids such complications. CONCLUSIONS: The novel self-expandable silicone airway stent may be a promising addition to commonly used stent types. Short-term and medium-term management of fistulas, tumor surface bleeding, and strictures (malignant and benign) is satisfactory. Scanning electron microscopy of stents provides information on peculiar features of microstructure and material that may be of use in clinical research and technical innovation. PMID- 9338638 TI - Relationship between early recurrence and micrometastases in the lymph nodes of patients with stage I non-small-cell lung cancer. AB - OBJECTIVE: This retrospective study was designed to detect occult micrometastases in the lymph nodes with the use of monoclonal anti-cytokeratin reagent, which is specific for epithelial cells but not for lymphocytes or plasmacytes, as well as to assess the relationship between the presence of occult micrometastases in the lymph nodes and an early relapse in patients with stage I non-small-cell lung cancer. METHODS: The paraffin-embedded sections of 973 regional lymph nodes from 44 patients with stage I non-small-cell lung cancer were studied. We used CAM-5.2 as the primary monoclonal anti-cytokeratin reagent and an indirect staining technique with the streptavidin-biotin-peroxidase complex method. RESULTS: We identified cytokeratin-positive cells in 31 (70.5%) of 44 patients and in 91 (9.4%) of the 973 lymph nodes. Of these 31 patients with cytokeratin-positive cells, 19 and 12 were restaged as having N1 and N2 disease, respectively. Thirteen patients had recurrent disease at 17 sites during the follow-up. Two of these recurrences were in the mediastinal nodes and the other 15 occurred at distant organs. Twelve of the 13 patients had micrometastatic disease in the regional lymph nodes. Disease-free survival duration was significantly shorter in the patients with micrometastases in the mediastinal lymph nodes than in patients with node-negative disease (p = 0.004). The independence of this prognostic significance was demonstrated by a multivariate analysis. CONCLUSION: These findings indicate that the detection of occult micrometastases in the mediastinal lymph nodes with monoclonal antibodies to cytokeratin can thus be used to predict an early relapse in patients with stage I non-small-cell lung cancer. PMID- 9338639 TI - Clinical experience in radical lymphadenectomy for adenocarcinoma of the gastric cardia. AB - OBJECTIVES: We evaluated the pattern of nodal metastasis and its prognosis after radical lymphadenectomy in adenocarcinoma of the gastric cardia. METHODS: We conducted a retrospective cohort study of 70 patients (52 men and 18 women; mean age 63.6 years) with adenocarcinomas of the gastric cardia who underwent extended gastrectomy (65 total gastrectomies and 5 proximal gastrectomies) and radical lymphadenectomy (D2 to D4) at Taichung Veterans General Hospital between 1989 and 1995. RESULTS: Twenty-four complications developed in 22 (31.4%) patients, and seven (10.0%) hospital deaths occurred. An overall 5-year cumulative survival of 37.6% was obtained. Lymph node metastases were identified in 53 (75.7%) patients. Nodal involvement was closely related to the depth of tumor invasion (p = 0.005). When the gastric wall invasion was limited to the subserosal layer (T1 and T2, n = 15), no patient had N4 group nodal metastasis. Once the serosal layer had been involved (beyond T3), N4 group nodal metastasis was frequently seen (30.9%, 17 of 55 patients). A multivariable analysis revealed that the level of nodal involvement, the depth of tumor invasion, and the presence of complications were independent prognostic factors. Cumulative 5-year survivals of curability A (n = 12), B (n = 19), and C (n = 32) resections were 100%, 21.2%, and 27.5%, respectively (p = 0.0001). The long-term survival of the patients after resection was also closely related to their pTNM stages (p = 0.0004). CONCLUSIONS: We conclude that gastrectomy accompanied by radical lymphadenectomy provides a reasonable long-term survival expectancy that is closely related to the stage of the disease and the curability of resection. PMID- 9338640 TI - The optimal Fontan connection: a growing extracardiac lateral tunnel with pedicled pericardium. AB - OBJECTIVE: The concept of a lateral tunnel for the Fontan operation is now widely accepted. Most lateral tunnels are constructed intraatrially with the use of aortic crossclamping. Construction of extracardiac lateral tunnels with the use of homografts or other nonviable tubes eliminates aortic crossclamping but lacks growth potential in length or width. The native pericardium, which is "sealed" posteriorly along the pulmonary artery, atrium, and inferior vena cava, could be turned down onto the right atrium to form a viable extracardiac lateral tunnel. METHODS: We designed and successfully constructed extracardiac lateral tunnels using viable autologous pericardium, pedicled on its lateral blood supply, in 19 patients aged 9 months to 5 years. All patients had a previous Glenn shunt; five patients had dextrocardia and a midline inferior vena cava. The patients' inferior vena cava-right atrial connection was opened transversely and the right atrial opening was sutured to its back wall, keeping the eustachian valve in the inferior vena cava. The underside of the right pulmonary artery was opened longitudinally; its inferior edge was sewn to the adjacent pericardial reflection. Any "pocket" or depressions in the posterior pericardium along the pulmonary veins were closed with running suture. Two incisions were made in the right pericardium down to the phrenic nerve parallel to the inferior vena caval and pulmonary arterial openings. This pedicled pericardium was trimmed and sewn as a roof to the upper edges of the inferior vena cava and pulmonary artery openings and then sewn longitudinally along the unopened right atrial wall, completing the viable extracardiac lateral tunnel. Although no fenestrations were used, these could be made during construction, or more significantly, owing to the lack of thick walled structures, in the catheterization laboratory in the postoperative period. RESULTS: All 19 patients had respiratory/cardiac pulsations in the pulmonary arteries owing to the compressible lateral tunnel. At follow-up of up to 2 1/2 years, all tunnels are growing and no obstructions have occurred. CONCLUSION: The viable autologous pericardial extracardiac lateral tunnel can be constructed without cardiac ischemia, can be fenestrated in the postoperative period, and forms a compressible, nonthrombogenic conduit capable of growth, which can be constructed early in infancy. PMID- 9338641 TI - Acute elevation of coronary venous pressure does not affect left ventricular contractility in the normal and stressed swine heart: implications for the Fontan operation. AB - OBJECTIVE: After the Fontan operation the right atrium and, thus, the coronary sinus are connected to the pulmonary arterial system, which causes the coronary venous pressure to increase. We investigated the acute effects of elevation of coronary venous pressure on baseline hemodynamics, coronary venous flow, and left ventricular contractility. METHODS: In acutely instrumented pigs, during complete right heart bypass and during constant cardiac output, pressure in the right atrium, right ventricle, and coronary sinus was altered by a height-adjustable reservoir. At various levels of coronary venous pressure (up to 4 kPa or up to 30 mm Hg), flow from the reservoir was measured and left ventricular hemodynamics and contractility were measured from catheter-derived left ventricular pressure and (conductance) volume data. Contractility of the left ventricle was assessed by the end-systolic pressure-volume relationship derived during an unloading intervention by adjusting the bypass pump speed. RESULTS: Left ventricular end diastolic pressure increased slightly (about 5%) with each kilopascal increase in coronary venous pressure, most likely related to diastolic ventricular interaction. No other changes in hemodynamic parameters occurred. Neither coronary venous flow nor left ventricular contractility was influenced by changes in coronary venous pressure. Imposing myocardial stress with dobutamine, 10 microg/kg per minute, did not change these findings. CONCLUSION: Increasing coronary venous pressure to 4 kPa in the intact circulation with intact autoregulation does not affect coronary flow or left ventricular contractility. We found no experimental evidence for the usefulness of diversion of the coronary sinus to the left atrium during Fontan-type operations PMID- 9338642 TI - The relation between pump flow rate and pulsatility on cerebral hemodynamics during pediatric cardiopulmonary bypass. AB - OBJECTIVES: Neurologic impairment, at least partly ischemic in origin, has been reported in up to 25% of infants undergoing cardiopulmonary bypass, with or without circulatory arrest. Controversy continues about the effect of pump flow, pulsatile or nonpulsatile, on the brain and in particular on cerebral blood flow. This study examines the relationship between pump flow rate and cerebral hemodynamics during pulsatile and nonpulsatile cardiopulmonary bypass. METHOD: Near-infrared spectroscopy was used to determine cerebral blood flow and cerebral blood volume (measured as concentration change) in a randomized crossover study. Pulsatile and nonpulsatile flow were used for six 5-minute intervals at each of three different pump flow rates (0.6, 1.2, and 2.4 L x m2 x min(-1)) in 40 patients, median age 2 months (range 2 weeks to 20 years 5 months). The relations between pulsatile flow, pump flow rate, cerebral blood flow, hemoglobin concentration change (cerebral blood volume), mean arterial pressure, arterial carbon dioxide tension, and hematocrit value were prospectively examined by means of multivariate analysis. RESULTS: Cerebral blood flow decreased 36% per L x m( 2) x min(-1) decrease in pump flow rate and was associated with changes in mean arterial pressure but did not differ according to pulsatility. Change in hemoglobin concentration was unrelated to changes in pulsatility of pump flow. CONCLUSION: Cerebral blood flow is related to pump flow rate. Pulsatile flow delivered with a Stockert pump does not increase cerebral blood flow or alter hemoglobin concentration during cardiopulmonary bypass in children. PMID- 9338643 TI - Cognitive and motor development in preschool and school-aged children after neonatal arterial switch operation. AB - OBJECTIVE: The developmental status of children beyond 3 years of age after the neonatal arterial switch operation has not yet been systematically evaluated and is the topic of the present work. METHODS: Seventy-seven unselected children operated on as neonates with combined deep hypothermic circulatory arrest and low flow cardiopulmonary bypass were examined at an age of 3.2 to 9.4 years (5.4 +/- 1.6 years, mean +/- standard deviation). Clinical neurologic status, standard scores of intelligence, acquired abilities and vocabulary, and standardized tests on gross motor and fine motor functions were carried out, and the results were related to preoperative, perioperative, and postoperative status and management. RESULTS: Neurologic impairment was more frequent (9.1%) than in the normal population. Intelligence was not different in these patients compared with normal children (p = 0.11), but motor function, vocabulary, and acquired abilities were poorer. Reduced intelligence was found in 9.1%, fine motor dysfunction in 22.1%, and gross motor dysfunction in 23.4% of the children. Intelligence was weakly but significantly inversely related to the duration of bypass (Spearman correlation coefficient -0.25, p = 0.03) and tended to be inversely related to the duration of circulatory arrest (-0.21, p = 0.07), but not to core cooling time on bypass or degree of hypothermia. Gross motor function, vocabulary, and acquired abilities were not significantly related to any of the perioperative parameters considered. No correlation was found between the test results and the variables perinatal asphyxia, perioperative and postoperative cardiocirculatory insufficiency, resuscitation events, and plexal or intraventricular cerebral hemorrhage. CONCLUSIONS: The neonatal arterial switch operation with combined circulatory arrest and low-flow bypass in our experience is associated with neurologic as well as fine and gross motor impairment but appears to be well tolerated concerning cognitive functions as based on formal intelligence testing. PMID- 9338644 TI - The left ventricular outflow tract in atrioventricular septal defect revisited: surgical considerations regarding preservation of aortic valve integrity in the perspective of anatomic observations. AB - OBJECTIVE: The anatomy of the left ventricular outflow tract in hearts with atrioventricular septal defect has been widely investigated, but controversies remain regarding detailed aspects of left ventricular outflow tract anatomy in the perspective of operative techniques to either prevent or relieve outflow tract obstruction. METHODS: We investigated 29 postmortem hearts with an atrioventricular septal defect. Measurements were taken of the circumferences and of the widths of the components that make up the outflow tract, that is, the interventricular septum, the superior bridging leaflet, the left ventricular free wall, and the length of the tendinous cords. RESULTS: The circumference of the left ventricular outflow tract immediately underneath the aortic valve was not different from that at the middle part of the outflow tract. Hearts with the partial type defect, characterized by separate atrioventricular orifices, had a smaller outflow tract than those with the complete variety. Although the anatomic constituents that contribute to left ventricular outflow tract obstruction are complex, this study showed that a reduced width of the interventricular septum was most intimately related to narrowing immediately underneath the aortic valve. Obstruction at the middle part of the left ventricular outflow tract was largely caused by reduced width of the interventricular septum together with short tendinous cords. CONCLUSIONS: On the basis of these observations, we recommend detailed investigation of the anatomy of the left ventricular outflow tract immediately underneath the aortic valve, before surgical attempts to relieve outflow tract obstruction, because in some procedures the integrity of the aortic valve will be at stake. PMID- 9338645 TI - The limits of detectable cerebral perfusion by transcranial Doppler sonography in neonates undergoing deep hypothermic low-flow cardiopulmonary bypass. AB - OBJECTIVE: Neurologic morbidity including seizures, abnormal neurologic function, and delayed psychomotor development continue to be significant problems for some patients undergoing operations for congenital heart disease, particularly for those subjected to deep hypothermic circulatory arrest. The technique of low-flow cardiopulmonary bypass has been advocated to decrease the incidence of neurologic sequelae. Our study examined the limits of detectable blood flow in the middle cerebral artery during low-flow cardiopulmonary bypass in 28 neonates undergoing the arterial switch procedure. METHODS: Cerebral blood flow velocity was measured noninvasively in the M1 segment of the middle cerebral artery with a 2 MHz range gated pulsed-wave transcranial Doppler sonographic probe that was placed over the left temporal window. As part of the initiation of a planned period of deep hypothermic circulatory arrest, the cardiopulmonary bypass flow rate was decreased in stages to five low-flow rates (50, 40, 30, 20, and 10 ml/kg per minute). After a period of stabilization, cerebral blood flow velocities were recorded at each of the five low-flow rates and reported as a percentage of baseline. RESULTS: All 28 neonates had detectable perfusion in the middle cerebral artery at flow rates of 30 ml/kg per minute or higher. At flows of 20 and 10 ml/kg per minute, one and eight, respectively, of the 28 neonates had no detectable perfusion in the middle cerebral artery. CONCLUSIONS: Our data show that cerebral perfusion can be detected by transcranial Doppler sonography in the middle cerebral artery in some neonates at bypass pump flows as low as 10 ml/kg per minute. However, when transcranial Doppler sonography was used in our patient population, a minimum bypass flow rate of 30 ml/kg per minute was needed to detect cerebral perfusion in all neonates. PMID- 9338646 TI - Influence of the pH of cardioplegic solutions on cellular energy metabolism and hydrogen ion flux during neonatal hypothermic circulatory arrest and reperfusion: a dynamic 31P nuclear magnetic resonance study in a pig model. AB - OBJECTIVES: The pH of cardioplegic solutions is postulated to affect myocardial protection during neonatal hypothermic circulatory arrest. Neither optimization of cardioplegic pH nor its influence on intracellular pH during hypothermic circulatory arrest has been previously studied in vivo. Thus we examined the effects of the pH of cardioplegic solutions on postischemic cardiac function in vivo, including two possible operative mechanisms: (1) reduction in adenosine triphosphate use and depletion of high-energy phosphate stores or (2) reduction of H+ flux during reperfusion, or both. METHODS: Dynamic 31P spectroscopy was used to measure rates of adenosine triphosphate use, high-energy phosphate depletion, cytosolic acidification during hypothermic circulatory arrest, and phosphocreatine repletion and realkalinization during reperfusion. Neonatal pigs in three groups (n = 8 each)--group A, acidic cardioplegia (pH = 6.8); group B, basic cardioplegia (pH = 7.8); and group N, no cardioplegia--underwent hypothermia at 20 degrees C with 60 minutes of hypothermic cardioplegia followed by reperfusion. RESULTS: Recoveries of peak elastance, stroke work, and diastolic stiffness were superior in group B. Indices of ischemic adenosine triphosphate use, initial phosphocreatine depletion rate, and tau, the exponential decay half time, were not different among groups. Peak [H+] in group A (end-ischemia) was significantly elevated over that of group B. The realkalinization rate was reduced in group B compared with that in groups A (p = 0.015) and N (p = 0.035), with no difference between groups A and N (p = 0.3). Cytosolic realkalinization rate was markedly reduced and the half-time of [H+] decay was increased during reperfusion in group B. CONCLUSIONS: Superior postischemic cardiac function in group B is not related to alterations in ischemic adenosine triphosphate use or high-energy store depletion, but may be due to slowing in H+ efflux during reperfusion, which should reduce Ca++ and Na+ influx. PMID- 9338648 TI - Transesophageal echocardiographic and clinical features of aortic intramural hematoma. AB - OBJECTIVE: This study sought to determine the transesophageal echocardiographic features and natural history of patients with aortic intramural hematoma. METHODS: The transesophageal echocardiograms of all patients who had symptoms indicative of aortic dissection over 6 years were reviewed. Measurements were made of the involved aortic segment in the study patients, and follow-up was obtained. RESULTS: In patients with aortic intramural hematoma, the wall thickness of the involved segment was significantly greater for descending segments than ascending segments (ascending aorta 7 +/- 2 mm, descending aorta 15 +/- 6 mm, p = 0.0016). In each case, the crescent-shaped intramural hematoma involved one wall predominantly, leading to compression of the aortic lumen. The findings of echolucent areas and displaced intimal calcium were found in the majority of patients. Four of eight patients with intramural hematoma of the ascending aorta were treated medically and four were treated surgically. The 30 day mortality was 50% in the medically treated patients and 0% in the surgically treated group. Four of 11 patients with isolated intramural hematoma of the descending aorta were treated medically and seven were treated surgically. All medically treated and 86% of surgically treated patients were alive at 30 days. CONCLUSIONS: Aortic intramural hematoma has distinct and identifiable transesophageal echocardiographic features. These data support those of previous studies documenting high morbidity and mortality in patients with aortic intramural hematoma. PMID- 9338647 TI - Protection of rat spinal cord from ischemia with dextrorphan and cycloheximide: effects on necrosis and apoptosis. AB - OBJECTIVE: We examined the characteristics of neuronal cell death after transient spinal cord ischemia in the rat and the effects of an N-methyl-D-aspartate antagonist, dextrorphan, and a protein synthesis inhibitor, cycloheximide. METHODS: Spinal cord ischemia was induced for 15 minutes in Long-Evans rats with use of a 2F Fogarty catheter, which was passed through the left carotid artery and occluded the descending aorta, combined with a blood volume reduction distal to the occlusion. The rats were killed after 1, 2, and 7 days. Other groups of rats were pretreated with dextrorphan (30 mg/kg, intraperitoneally, n = 7), cycloheximide (30 mg, intrathecally, n = 7), or vehicle (saline solution, n = 12) and killed after 2 days. RESULTS: This model reliably produced paraplegia and histopathologically distinct morphologic changes consistent with necrosis or apoptosis by light and electron microscopic criteria in different neuronal populations in the lumbar cord. Scattered necrotic neurons were seen in the intermediate gray matter (laminae 3 to 7) after 1, 2, and 7 days, whereas apoptotic neurons were seen in the dorsal horn laminae 1 to 3 after 1 and 2 days. Deoxyribonucleic acid extracted from lumbar cord showed internucleosomal fragmentation (laddering) on gel electrophoresis indicative of apoptosis. The severity of paraplegia in the rats treated with dextrorphan and cycloheximide was attenuated 1 day and 2 days after ischemia. The numbers of both necrotic and apoptotic neurons were markedly reduced in both dextrorphan- and cycloheximide treated rats. CONCLUSIONS: The results suggest that both N-methyl-D-aspartate receptor-mediated excitotoxicity and apoptosis contribute to spinal cord neuronal death after ischemia and that pharmacologic treatments directed at blocking both of these processes may have therapeutic utility in reducing spinal cord ischemic injury. PMID- 9338649 TI - Reevaluating the significance of pulmonary hypertension before cardiac transplantation: determination of optimal thresholds and quantification of the effect of reversibility on perioperative mortality. AB - OBJECTIVES: Right-sided circulatory failure resulting from severe preoperative pulmonary hypertension is a source of mortality early after cardiac transplantation. We undertook the present study (1) to analyze the association of elevated pulmonary hemodynamic indices with 30-day mortality, (2) to define threshold ranges associated with an increase in 30-day mortality, and (3) to evaluate the effect of vasodilator reversibility on 30-day mortality. METHODS: Preoperative hemodynamic profiles were evaluated in 476 patients who ultimately underwent cardiac transplantation. From these data, receiver-operating characteristic curves and stratum-specific likelihood ratios were generated to compare the efficacy of each hemodynamic index. A subset of patients with elevated hemodynamic profiles at baseline additionally underwent graded sodium nitroprusside infusion. RESULTS: Analysis of receiver-operating characteristic curves demonstrated no statistically significant difference among the indices in their ability to predict 30-day mortality. Analysis of stratum-specific likelihood ratios demonstrated three risk strata that correlated with significant differences in 30-day mortality, with patients in the high-risk stratum having a 3.2 to 4.4 increase in odds of 30-day mortality when compared with patients in the low-risk stratum. Nitroprusside data demonstrated that although 30-day mortality was better in patients with reversible pulmonary hypertension than in those with fixed pulmonary hypertension, it was not comparable with that of patients without pulmonary hypertension at baseline. CONCLUSIONS: Candidates for cardiac transplantation may be categorized into three risk strata on the basis of their preoperative pulmonary hemodynamic profile; the association of this profile with 30-day mortality is not linear. Reversibility with nitroprusside appears to confer some improvement in the risk of 30-day mortality, but it may not eliminate the risk entirely. PMID- 9338650 TI - Dibutyryl cyclic adenosine monophosphate attenuates lung injury caused by cold preservation and ischemia-reperfusion. AB - OBJECTIVE: Dibutyryl adenosine 3',5'cyclic monophosphate (db-cAMP) is a membrane permeable analog of adenosine 3',5'cyclic monophosphate (cAMP). We examined the effect of db-cAMP against lung injury caused by cold preservation and ischemia reperfusion. METHODS: Rats were divided into three groups (each n = 6) according to the presence or absence of db-cAMP in the preservative solution and cold ischemia (4 degrees C for 15 hours). In the fresh group, the lung was flushed with the preservative solution and reperfusion was performed immediately. In the control group and the db-cAMP group, the lung was flushed either with the solution or with a combination of the solution plus db-cAMP, respectively, and preserved at 4 degrees C for 15 hours. The lung was reperfused for 60 minutes in an ex vivo rat lung perfusion model. RESULTS: The shunt ratios of the reperfused lung in the db-cAMP group were 4.0% +/- 1.6% and 3.4% +/- 1.2% 10 and 60 minutes, respectively, after the initiation of reperfusion, being as low as those in the fresh group and significantly lower than those in the control group (p < 0.01). The wet/dry weight ratio of the lung tissue after reperfusion was 5.99 +/- 1.50 in the db-cAMP group, which was similar to that in the fresh group (5.45 +/- 0.23) and significantly lower than that in the control group (14.20 +/- 3.43) (p < 0.01). Electron microscopic examination showed less damage in the pulmonary arterial endothelium in the db-cAMP group. CONCLUSIONS: We conclude that db-cAMP attenuates the lung injury by cold preservation and ischemia-reperfusion, at least partly by protection of the vascular endothelium. PMID- 9338652 TI - Protein kinase C activation before cardioplegic arrest: beneficial effects on myocyte contractility. AB - OBJECTIVE: A potential intracellular mechanism for the protective effects of myocardial preconditioning is the activation of protein kinase C. The present study tested the hypothesis that a brief period of protein kinase C activation before cardioplegic arrest would provide protective effects on myocyte contractility with subsequent reperfusion and rewarming. METHODS: Left ventricular porcine myocytes were assigned to the following treatments: (1) Protein kinase C/cardioplegia: Protein kinase C activation in myocytes (n = 39) for 3 minutes with a phorbol ester (10(-9) mol/L of phorbol 12-myristate 13 acetate) in oxygenated, normothermic (37 degrees C) cell media. Protein kinase C activation was followed by 2 hours of cardioplegic arrest (K+, 24 mEq/L; HCO3-, 30 mEq/L; 4 degrees C) and a 5-minute reperfusion period (37 degrees C media). (2) Cardioplegia: Myocytes (n = 31), 2 hours of cardioplegic arrest, and a 5 minute reperfusion and rewarming period. Myocyte contractility was measured by means of high-speed videomicroscopy. For comparison purposes, contractile function was examined in myocytes (n = 70) under normothermic control conditions. RESULTS: Myocyte shortening velocity was reduced after cardioplegic arrest when compared with normothermic values (22.3 +/- 1.6 vs 48.8 +/- 2.0 microm/sec, p < 0.0001). Protein kinase C activation before cardioplegic arrest normalized myocyte shortening velocity (48.8 +/- 2.5 microm/sec). Co-incubation with phorbol 12-myristate 13-acetate and chelerythrine (10(-6) mol/L), an inhibitor of protein kinase C, before cardioplegic arrest abolished the protective effects of phorbol 12-myristate 13-acetate pretreatment. CONCLUSION: These results suggest that an endogenous means of providing improved myocardial protection during prolonged cardioplegic arrest can be achieved through a brief period of protein kinase C activation. PMID- 9338651 TI - Superiority of hyperpolarizing to depolarizing cardioplegia in protection of coronary endothelial function. AB - OBJECTIVE: Hyperpolarizing cardioplegia has recently been proposed for myocardial protection. To compare the protective effect of hyperpolarizing cardioplegia and depolarizing (hyperkalemic) cardioplegia on coronary endothelium, we studied porcine coronary arteries in the organ chamber. METHODS: Relaxation mediated by the endothelium-derived hyperpolarizing factor (EDHF) was used as the index of endothelial function because (1) hyperkalemia without ischemia does not impair the nitric oxide-mediated function according to previous studies and (2) EDHF relaxes vessels by hyperpolarizing the membrane potential. Therefore depolarizing cardioplegia may inhibit this function, but hyperpolarizing cardioplegia may preserve it. EDHF-mediated relaxation was induced by bradykinin and the calcium ionophore A23187 with the presence of indomethacin (7 micromol/L; INN: indometacin), a cyclooxygenase inhibitor, and N(G)-nitro-L-arginine (300 micromol), a nitric oxide biosynthesis inhibitor in U46619 (30 nmol/L)-induced precontraction. The vessels were exposed to either hyperpolarizing cardioplegic solution (the potassium-channel opener aprikalim, 0.1 mmol/L) or depolarizing cardioplegic solution (high potassium concentration, 20 mmol/L for A23187 and 50 mmol/L for bradykinin experiments) for 1 hour with a constant supply of oxygen to exclude the effect of ischemia. RESULTS: EDHF-mediated relaxation was significantly impaired in either A23187 or bradykinin studies (80.1% +/- 7.5% vs 24.9% +/- 14.2%, p = 0.004, n = 8 in each group for A23187, and 71.4% +/- 4.7%, n = 13, vs 40.5% +/- 12.9%, n = 7, p = 0.01, for bradykinin). The effective concentration causing 50% of maximal relaxation was significantly increased in the A23187 experiments with the treatment of hyperkalemia. In contrast, in aprikalim-treated arteries, the EDHF-mediated relaxation induced by either A23187 or bradykinin was unchanged. CONCLUSIONS: We conclude that EDHF-mediated coronary endothelial function is maximally preserved by hyperpolarizing cardioplegia but impaired by depolarizing cardioplegia. These findings support the use of hyperpolarizing cardioplegia in cardiac operations. PMID- 9338653 TI - Retrograde cerebral perfusion provides limited distribution of blood to the brain: a study in pigs. AB - OBJECTIVE: The objective of this study was to investigate flow distribution during retrograde and antegrade cerebral perfusion with India ink as a marker. METHODS: Ten pigs received cerebral perfusion with a solution containing 50% filtered India ink for 5 minutes either antegradely through both internal carotid arteries at a flow of 180 to 200 ml/min (n = 5) or retrogradely via the superior vena cava at a flow of 300 to 500 ml/min (n = 5). The brains were then fixed for quantitative measurement of the density of ink-filled capillaries (reported as a percentage of the total selected area). The assessment was done with the use of an in-house software program. RESULTS: In the antegrade cerebral perfusion group, the intracranial arterial and venous systems were completely filled with ink. The gray matter was colored uniformly black, and light coloring was observed in the white matter. During retrograde cerebral perfusion, the majority of ink was returned to the inferior vena cava, and only a small amount of ink was found in the innominate artery draining from the brain. Massive ink filling was observed in the sagittal sinus and other venous sinuses in all the pigs. Vessels on the surface of the brain and large vessels in the brain were also well filled with ink. However, only 10% of capillaries were filled with ink during retrograde cerebral perfusion relative to the number observed with antegrade cerebral perfusion. CONCLUSIONS: Retrograde cerebral perfusion supplies a limited amount of blood to brain tissue, which flows mainly through superficial and large deep cerebral vessels. PMID- 9338654 TI - Natriuresis after cardiopulmonary bypass: relationship to urodilatin, atrial natriuretic factor, antidiuretic hormone, and aldosterone. AB - OBJECTIVE: Recent studies suggest that urodilatin from the kidneys rather than atrial natriuretic factor from the heart is the more important member of the family of natriuretic peptides involved in the normal regulation of renal sodium and water excretion. We thus examined the relationship between natriuresis, urodilatin, and atrial natriuretic factor in patients after cardiopulmonary bypass, a procedure known to increase levels of atrial natriuretic factor significantly. METHODS: Excretion rates of sodium and water were correlated with the excretion of urodilatin and with circulating levels of atrial natriuretic factor, antidiuretic hormone, aldosterone, and plasma renin activity during a period of 16 hours in 12 patients having had coronary artery bypass operations and with approximately a 400% elevation in levels of atrial natriuretic factor. RESULTS: Natriuresis did not correlate with atrial natriuretic factor, antidiuretic hormone, aldosterone, or plasma renin activity. Excretion rates of urodilatin, however, correlated significantly with excretion rates of sodium (r = 0.74, p = 0.03), urine flow (r = 0.83, p = 0.01), and with levels of serum sodium (r = 0.82, p = 0.01). CONCLUSION: These results suggest an important role for urodilatin, greater than that of atrial natriuretic factor, in the regulation of renal excretion of sodium and water after cardiopulmonary bypass surgery. PMID- 9338655 TI - Extracardiac myxoma: an unusual right ventricular epicardial location. PMID- 9338656 TI - Cardiac tamponade caused by penetration of an acupuncture needle into the right ventricle. PMID- 9338657 TI - Long-term follow-up of aprotinin-specific immunoglobulin G antibodies after cardiac operations. PMID- 9338658 TI - Primary tracheal synovial cell sarcoma: a first case report. PMID- 9338659 TI - Immobilized instrument for minimally invasive direct coronary artery bypass: MIDCAB doughnut. PMID- 9338660 TI - Minimally invasive redo aortic valve replacement. PMID- 9338661 TI - Primary immunoblastic B-cell lymphoma of the sternum. PMID- 9338662 TI - Partial sternotomy: a mini approach for maxi exposure. PMID- 9338663 TI - The right internal thoracic artery for myocardial revascularization. PMID- 9338664 TI - Early tracheal extubation after coronary artery bypass grafting. PMID- 9338665 TI - The safety of peritoneal drainage and dialysis after cardiopulmonary bypass in children. PMID- 9338666 TI - Effect of pterygium morphology on pterygium recurrence in a controlled trial comparing conjunctival autografting with bare sclera excision. AB - OBJECTIVES: To compare success rates of conjunctival autografting and bare sclera excision for primary and recurrent pterygium in the tropics and to evaluate risk factors for pterygium recurrence. METHODS: A prospective, controlled clinical trial was performed in which 123 primary and 34 recurrent pterygia, matched for age and pterygium morphology, were randomized in 2 separate studies to receive either bare sclera excision or conjunctival autograft. The surgical procedures were performed by one surgeon and reviewed at 1, 3, 6, and 12 months after surgery by an independent observer. Pterygium morphology was clinically graded as atrophic, intermediate, or fleshy according to an assessment of pterygium translucency. Risk factors were assessed using likelihood ratio tests. Weibull curves were used to estimate recurrence rates allowing for the interval censoring. RESULTS: In the group with primary pterygium (mean follow-up, 15.1 months), 38 (61%) of the 62 cases of bare sclera excision (heretofore referred to as the bare sclera group) had pterygium recur in contrast with 1 (2%) of the 61 cases of conjunctival autograft (heretofore referred to as the conjunctival autograph group) (P<.001, likelihood ratio X2 test). Nontranslucency, or fleshiness of the pterygium, and not age was a significant risk factor for recurrence in the bare sclera group (P<.001, likelihood ratio X2 test). In the group with recurrent pterygium (mean follow-up, 13.2 months), 14 (82%) of the 17 bare sclera group had pterygium recur, while no recurrences occurred among 17 cases in the conjunctival autograft group. Nontranslucency was again a highly significant factor for recurrence (P<.001, likelihood ratio X2 test). CONCLUSIONS: Pterygium recurrence is related to pterygium morphology and fleshiness of the pterygium is a significant risk factor for recurrence if bare sclera excision is performed. Conjunctival autografting for primary and recurrent pterygium is effective in reducing pterygium recurrence compared with bare sclera excision. PMID- 9338667 TI - Topical 2.0% dorzolamide vs oral acetazolamide for prevention of intraocular pressure rise after neodymium:YAG laser posterior capsulotomy. AB - OBJECTIVE: To compare the efficacy and safety of topical 2.0% dorzolamide hydrochloride with oral acetazolamide in preventing intraocular pressure (IOP) rise following neodymium:YAG (Nd:YAG) laser posterior capsulotomy. DESIGN: A prospective, randomized, double-masked, placebo-controlled study. PATIENTS: Two hundred ten patients undergoing Nd:YAG laser posterior capsulotomy. INTERVENTION: Pretreatment with dorzolamide, acetazolamide, or placebo. Dorzolamide administration as a single drop (1 drop approximately 20 microL) 1 hour before capsulotomy. Acetazolamide administration as a single dose of 125 mg orally 1 hour before capsulotomy. RESULTS: At first and third hour postoperatively, IOPs and IOP changes from baseline were significantly (P<.001) higher in the placebo group than in the dorzolamide or acetazolamide group. At the same time, IOPs and IOP changes from baseline were similar (P>.50) in the dorzolamide and acetazolamide groups. No patient treated with dorzolamide or acetazolamide experienced an IOP higher than 30 mm Hg after capsulotomy, but 15.7% of patients receiving placebo had an IOP above this level (P<.001). Of patients receiving placebo, 5.7% experienced IOP higher than 35 mm Hg. No serious side effects were recorded in any of the studied patients. CONCLUSION: Topical 2.0% dorzolamide and oral acetazolamide, given prophylactically as a single administration 1 hour before Nd:YAG laser posterior capsulotomy, have comparable high efficacy and safety in preventing IOP elevation following this procedure. PMID- 9338669 TI - Molteno implantation for secondary glaucoma in juvenile rheumatoid arthritis. AB - OBJECTIVE: To evaluate the outcome of Molteno implantation in secondary glaucoma associated with juvenile rheumatoid arthritis. METHODS: Between January 1, 1986, and December 1, 1996, 27 eyes of 19 consecutive patients with secondary glaucoma due to juvenile rheumatoid arthritis received a Molteno implant. The diagnosis of juvenile rheumatoid arthritis was made according to the American Rheumatism Association criteria. RESULTS: At the end of the follow-up (mean, 40 months; range, 6-116 months), the mean (+/-SD) postoperative intraocular pressure (IOP) (14.4+/-4.3 mm Hg) was statistically significantly lower than the preoperative IOP (38.3+/-5.6 mm Hg) (P<.001). The Snellen visual acuity remained within 1 line of the preoperative level or improved in 23 (85%) of 27 eyes. A successful outcome (defined as a final IOP of > or =6 mm Hg and < or =22 mm Hg, with fewer than or an equal number of antiglaucoma medications as preoperatively) was achieved in 24 (89%) of 27 eyes. Life-table analysis success rates were 95% after 27 months and 90% after 52 months of follow-up. Postoperative complications included flat anterior chamber (3 eyes), tube block by iris or vitreous (3 eyes), cataract (3 eyes), cornea-tube touch (2 eyes), choroidal detachment (1 eye), corneal edema (1 eye), and corneal abrasion (1 eye). CONCLUSION: The Molteno implant is useful and well tolerated in controlling IOP in patients with glaucoma secondary to juvenile rheumatoid arthritis. PMID- 9338668 TI - Combined phacoemulsification and filtering surgery with the 'no-stitch' technique. AB - OBJECTIVE: To determine if intentionally making a radial incision could lead to a lasting decrease in intraocular pressure and the development of filtering blebs. METHODS: Forty-three eyes treated with combined filtering surgery were compared with a control group of 42 eyes treated with cataract surgery (phacoemulsification) alone. All of the patients had advanced chronic open-angle glaucoma. During combined cataract and glaucoma surgery, the tunnel floor was transected with Vannas scissors. RESULTS: One year after surgery, the mean intraocular pressure in the study group decreased 7.6 mm Hg from a preoperative mean (+/-SD) of 25.9+/-5.3 mm Hg. The decrease in the control group was 3.7+/-4.2 mm Hg. The difference between the 2 groups was statistically significant (P<.001). In the study group, an average of 1.5+/-0.8 fewer medications were required 1 year after surgery. In the control group, 0.5+/-0.6 fewer medications were required. CONCLUSIONS: The combined surgical procedure discussed in this article led to a lasting decrease in intraocular pressure. Cataract surgery alone using the no-stitch technique and posterior chamber lens implantation also reduced intraocular pressure, although significantly less. PMID- 9338670 TI - Deterioration of visual fields in patients with glaucoma with and without optic disc hemorrhages. AB - OBJECTIVE: To evaluate visual field deterioration in patients with glaucoma with and without optic disc hemorrhages (DHs). DESIGN: A prospective study at quarterly base involving annual perimetry; mean follow-up of 9 years. SETTING: Outpatient department, nonreferral basis. PATIENTS: Sixty-eight patients with primary open-angle glaucoma, 34 with normal pressure glaucoma (NPG), and 125 with ocular hypertension. RESULTS: Visual field deterioration occurred in 32%, 32%, and 6% of the patients without DHs who had NPG, primary open-angle glaucoma, or ocular hypertension, respectively, while visual field deterioration occurred in 80%, 89%, and 14% of patients with DH, respectively. Cox proportional hazards ratio(CHR) for deterioration in patients with vs patients without DHs was 5.4 for NPG (P<.01) and 3.6 for primary open-angle glaucoma (P<.01). In patients with NPG and DHs, ipsilateral eyes with DHs deteriorated in 58%, while contralateral eyes without DHs deteriorated in 11% (CHR, 8.9; P<.04). For primary open-angle glaucoma and ocular hypertension, progression did not differ between eyes with DHs and contralateral eyes without DHs. Mean (+/-SD) interval between DHs and ipsilateral visual field deterioration was 3.1+/-1.7 years. No difference in the proportion of eyes progressing after single or recurrent DHs was noted. The position of DHs was related to the site of the visual field loss in 44% of the eyes. CONCLUSIONS: The presence of DHs increased the risk of visual field deterioration. Disc hemorrhages were indicative only of deterioration in ipsilateral eyes in patients with NPG. PMID- 9338672 TI - Transscleral vs transpupillary diode laser photocoagulation for the treatment of threshold retinopathy of prematurity. AB - OBJECTIVE: To evaluate the efficacy and safety of transscleral diode laser photocoagulation for the treatment of threshold retinopathy of prematurity (ROP). PATIENTS: Fifty eyes of 25 preterm infants (birth weight, 510-1200 g [864+/-178 g ?mean+/-SD?]; gestational age, 24-29 weeks [26.7+/-1.7 weeks]) with threshold ROP were treated with diode laser photocoagulation (wavelength, 810 nm). One eye of each infant was treated transsclerally while the fellow eye had transpupillary coagulation using the laser indirect ophthalmoscope. Follow-up ranged from 2 to 22 months (10.0+/-5.3 months). MAIN OUTCOME MEASURE: The regression of acute ROP and the incidence of adverse treatment effects. RESULTS: In 25 (100%) of the eyes treated transpupillarly and in 24 (96%) of the eyes treated transsclerally, ROP regressed after a single or a second laser treatment and the outcome was a flat, attached retina. One eye (4%) with disease in zone I failed to improve after transscleral laser treatment and ROP progressed to stage 4B with a partially attached retina, although additional retinal detachment surgery with an encircling band was performed. No adverse side effects as a result of diode laser treatment were noted except for a small amount of retinal-preretinal bleeding in the ridge in 9 (36%) of the transsclerally and in 5 (20%) of the transpupillarly coagulated eyes. There were no adverse side effects (eg, cataract formation) in the anterior segments of the eyes. CONCLUSIONS: The results suggest that transscleral diode laser coagulation is as effective in the treatment of threshold ROP as transpupillary diode laser photocoagulation. Only minor side effects were noted. Transscleral diode laser photocoagulation seems to be an advantageous treatment method if transpupillary treatment bears an increased risk of cataract formation. PMID- 9338671 TI - Large cups in normal-sized optic discs: a variant of optic nerve hypoplasia in children with periventricular leukomalacia. AB - OBJECTIVE: To evaluate optic disc morphologic features in children with periventricular leukomalacia (PVL). PATIENTS AND METHODS: Seventeen children with PVL (patient group) were compared with 17 sex- and age-matched, full-term healthy control children (control group). Clinical ophthalmological examination and digital image analysis of fundus photographs were performed in all children. In children with PVL, cerebral imaging was performed. RESULTS: Children with PVL had larger optic disc cupping (P<.02) than did control children. A large proportion of the children with PVL had strabismus, nystagmus, and restricted visual fields. CONCLUSION: Our study indicates that optic nerve hypoplasia in children with PVL is often associated with a normal-sized optic disc with a large cup. This unusual form of optic nerve hypoplasia most likely results from transsynaptic degeneration of optic axons caused by the primary bilateral lesion in the optic radiation. PMID- 9338673 TI - Membranectomy and autologous serum for the retreatment of full-thickness macular holes. AB - OBJECTIVE: To determine the efficacy of reoperation with rigorous epiretinal membrane dissection and autologous serum for full-thickness macular holes remaining open after initial surgery. METHODS: Forty-six consecutive eyes that had previously undergone unsuccessful macular hole surgery were re-treated with epiretinal membrane dissection, adjunctive autologous serum, and 16% perfluoropropane (C3F8) gas tamponade. Anatomical closure and improvement of best corrected Snellen visual acuity were used as outcome measures, and nuclear sclerosis was graded clinically before and after reoperation. RESULTS: Epiretinal membrane was identified and dissected in 29 (63%) of the 46 eyes and anatomical closure was achieved in 37 (80%) of the 46 eyes. Of these, 23 (62%) of 37 improved by at least 2 Snellen lines, 12 (35%) of 37 by at least 3 Snellen lines, and 6 (16%) of 37 by at least 4 Snellen lines. Increase in nuclear sclerosis occurred in 30 (65%) of the 46 eyes postoperatively, leading to cataract extraction in 12 (26%) of the eyes at last follow-up (mean, 10.3 months). A longer total duration (P<.001) and a worse preoperative visual acuity (P=.001), prior to reoperation, were associated with a worse final visual acuity after surgery. CONCLUSIONS: Retreatment with rigorous membranectomy and autologous serum seems to be beneficial in most eyes in which initial macular hole surgery has failed. Although the anatomical closure rate is similar to that reported after primary surgery, final visual acuity improvement seems to be less than after successful primary closure, owing to the longer mean duration of holes in which initial surgery has failed. PMID- 9338674 TI - Contractile responses of cultured bovine retinal pericytes to angiotensin II. AB - OBJECTIVE: To document that angiotensin (ANG) II contracts cultured bovine retinal pericytes via saralasin-sensitive receptors if the cells are prerelaxed. METHODS: Changes in the contractile tone were quantified as the changes in the summed length of wrinkles induced by pericytes cultured on the silicone surface. RESULTS: Angiotensin II (10(-5) mol/L) did not increase the contractile tone of cultured pericytes that were not prerelaxed. However, when the pericytes had been prerelaxed 41% with 10(-6)-mol/L sodium nitroprusside, ANG II at the range of 10( 7) to 10(-5) mol/L caused prompt, dose-related, significant (P<.01) contraction. It induced a maximum contraction (29.9%+/-5.2% [mean+/-SE]) at 10(-6) mol/L. This effect lasted at least 10 minutes. Angiotensin II receptor antagonist saralasin (10(-6) mol/L) abolished the contractile effect of ANG II (10(-6) mol/L), although by itself it did not affect the contractile tone. CONCLUSIONS: Angiotensin II contracts cultured pericytes through saralasin-sensitive ANG II receptors. If ANG II affects the contractile tone of pericytes in vivo, it may affect capillary caliber, resistance, and blood flow. PMID- 9338675 TI - Subconjunctival carboplatin therapy and cryotherapy in the treatment of transgenic murine retinoblastoma. AB - OBJECTIVES: To determine the efficacy and dose response of subconjunctival carboplatin with and without cryotherapy in the treatment of murine transgenic hereditary retinoblastoma. METHODS: Fifty-one 5-week-old transgenic BLH SV-40 (Charles Rivers Laboratories, Boston, Mass) T-antigen-positive mice with retinoblastoma were administered 6 subconjunctival injections of carboplatin in 1 eye at drug doses of 10, 15, 20, 25, 62.5, 125, and 250 microg. Six control eyes received 6 subconjunctival injections of balanced salt solution. Fourteen of the 51 subconjunctivally treated eyes received a single application of transconjunctival cryotherapy immediately prior to each carboplatin injection. Six control eyes received 6 single applications of transconjunctival cryotherapy using the above schedule but did not receive carboplatin. All experimental and control eyes were obtained at 16 weeks of age for histopathologic examination. RESULTS: A dose-dependent inhibition of intraocular tumor growth by subconjunctivally delivered carboplatin was observed in these transgenic retinoblastoma mice. Tumor development was inhibited in 50% of the mouse eyes at doses of 180 microg. In animals treated with cryotherapy alone, no tumor control was noted (0 of 6). In animals treated with subconjunctival carboplatin coupled with cryotherapy, a tumor control dose of 417 microg was found. No evidence of histopathologic treatment toxicity was noted. CONCLUSIONS: Subconjunctival delivery of carboplatin in serial doses effectively inhibits intraocular tumor growth in a dose-dependent fashion in a transgenic murine retinoblastoma model. Cryotherapy does not increase tumor control in this murine retinoblastoma model. PMID- 9338676 TI - HIV and AIDS and the eye in developing countries: a review. AB - An estimated 30 million people worldwide have been infected with the human immunodeficiency virus, the causative agent of the acquired immunodeficiency syndrome. Of these, 90% live in developing countries from where there is relatively little published information about the ocular manifestations of human immunodeficiency virus and acquired immunodeficiency syndrome. We review the information available from Africa, Latin America, and Asia. The prevailing ocular manifestations differ in some developing countries compared with those in the industrialized countries. These differences most likely result from different socioeconomic conditions and basic health care availability and from different patterns of endemic disease present before the human immunodeficiency virus epidemic. PMID- 9338677 TI - Alcohol, smoking, and cataracts: the Blue Mountains Eye Study. AB - OBJECTIVE: To investigate the associations between alcohol consumption, tobacco smoking, and cataract. DESIGN: A population-based, cross-sectional study. SETTING: An urban community in the Blue Mountains, close to Sydney, Australia. PARTICIPANTS: Three thousand six hundred fifty-four people aged 49 to 97 years. The participation rate was 82%. MAIN OUTCOME MEASURES: Smoking history and details of current alcohol consumption were assessed by questionnaire. Lens photographs were taken and graded for presence and severity of cortical, nuclear, and posterior subcapsular cataracts. RESULTS: After adjusting for multiple potential confounders, people who had ever smoked cigarettes had a higher prevalence than nonsmokers of more severe nuclear (adjusted odds ratio [OR], 1.3; 95% confidence interval [CI], 1.1-1.6) and posterior subcapsular (adjusted OR, 1.5; 95% CI, 1.1-2.1) cataracts. The association between pipe smoking and nuclear cataract (adjusted OR, 3.1; 95% CI, 1.5-8.2) was stronger than the association with cigarette smoking. Alcohol consumption was associated with a reduced prevalence of cortical cataract: compared with people who did not drink, the adjusted OR for cortical cataract among people who drank at least 1 drink a day was 0.7 (95% CI, 0.6-0.9). Heavy alcohol consumption (> or =4 drinks a day) was associated with nuclear cataract in current smokers (adjusted OR compared with nondrinkers, 3.9; 95% CI, 0.9-16.6) but not in never smokers. CONCLUSIONS: Consistent with other studies, smoking was associated with a higher prevalence of nuclear and posterior subcapsular cataracts. The only adverse effect of alcohol was among smokers: people who smoked and drank heavily had an increased prevalence of nuclear cataract. PMID- 9338678 TI - International variation in anesthesia care during cataract surgery: results from the International Cataract Surgery Outcomes Study. AB - OBJECTIVES: To describe international variation in anesthesia care and monitoring during cataract surgery and to discuss its implications for cost and safety. METHODS: A standardized questionnaire was sent to random samples of ophthalmologists in the United States, Canada, and Barcelona, Spain, and to all ophthalmologists in Denmark. The survey was conducted in 1993 and 1994. Certified ophthalmologists who had performed 1 or more cataract extractions in the previous year were eligible for enrollment. RESULTS: The response rates were 62% in the United States (n=148), 67% in Canada (n=276), 70% in Barcelona (n=89), and 80% in Denmark (n=82). The anesthetic technique for cataract surgery varied significantly between sites (P<.001). Surgeons reported that retrobulbar blocks were used for 46% of the cataract extractions in the United States, 70% in Canada, 66% in Denmark, and 31% in Barcelona. In Barcelona, general anesthesia was used for 23% of the cataract extractions; it was used for less than 3% of the extractions at the other 3 sites. Peribulbar blocks or topical anesthesia was used for the remaining extractions. In the United States, Canada, and Barcelona, surgeons reported that vital functions were monitored during more than 97% of the extractions and anesthesia surveillance was used during more than 78% of the extractions. In Denmark, ophthalmologists reported that vital functions were monitored and anesthesia surveillance was used for 1% of the cataract extractions (P<.001). CONCLUSIONS: Substantial international variation in anesthesia care and monitoring during cataract surgery was observed. The findings suggest a need for further research to determine whether less intensive monitoring is cost effective. PMID- 9338679 TI - What is the future of research in age-related macular disease? PMID- 9338680 TI - The ARCHIVES and Japanese ophthalmology. PMID- 9338682 TI - Paecilomyces lilacinus endophthalmitis with secondary keratitis: a case report and literature review. AB - A case of endogenous fungous endophthalmitis with secondary pupillary block glaucoma and corneal invasion requiring penetrating keratoplasty is reported. Initially Paecilomyces lilacinus was isolated from a vitreous and a lens aspirate, but a second vitreous tap revealed Aspergillus fumigatus and P lilacinus. This case highlights the difficulty of diagnosing endogenous fungous endophthalmitis presenting without risk factors and the difficulties of managing such cases using the antifungous agents available. To our knowledge, this is the first case report documenting a progression to stromal keratitis from endogenous endophthalmitis secondary to P lilacinus. PMID- 9338681 TI - Mycobacterium chelonae keratitis after excimer laser photorefractive keratectomy. AB - This is the first report of a severe case of Mycobacterium chelonae keratitis; it occurred in a 26-year-old man after he had undergone excimer laser photorefractive keratectomy for the correction of severe myopia, once the epithelium was already healed. The diagnosis was made by culture results and acid fast staining of corneal scrapings. Topical ciprofloxacin sodium, 0.3 mg/mL, plus amikacin sodium, 10 mg/mL, and oral clarithromycin sodium led to remission of the ulceration after 3 months of therapy. Subsequent topical corticosteroid therapy led to complete visual recovery during 1 year of follow-up. There may be an increased risk of severe keratitis during the first postoperative months in eyes that have already undergone photorefractive keratectomy, due to the presence of some microepithelial defects symptomatically negative and not easily detectable by slit-lamp examination. PMID- 9338683 TI - Medulloepithelioma of the optic nerve head. AB - Medulloepithelioma of the optic nerve is a rare developmental tumor. We describe a 2-year-old boy with profound loss of vision associated with a visible tumor of the optic nerve head in his left eye. A clinically diagnosed retinoblastoma necessitated left eye enucleation. The histopathological diagnosis was malignant medulloepithelioma that was incompletely resected. Further tumor resection was required, and the patient received adjunctive chemotherapy and radiotherapy. Four years after treatment, the patient has neither clinical nor radiological evidence of tumor. PMID- 9338684 TI - Congenital communication of cilioretinal and retinal arteries associated with angioid streaks. PMID- 9338685 TI - Acquired prepapillary vascular loops. PMID- 9338686 TI - Esthesioneuroblastoma presenting with epiphora in a young child. PMID- 9338687 TI - Ocular neuromyotonia 18 years after radiation therapy. PMID- 9338688 TI - The double-mirror gonioscopic lens for surgery of the anterior chamber angle. AB - Intraoperative examination of the anterior chamber angle is necessary during several glaucoma procedures, including goniosynechialysis and goniotomy. Many of the available lenses for this purpose are difficult to use because they require oblique illumination and their size interferes with surgical manipulations at the limbus. We have developed a double-mirror gonioscopic lens that uses direct illumination; provides an upright image of angle structures; and has a small diameter, allowing simultaneous access to the entire limbal region. We compared our lens with the Posner goniomirror (Ocular Instruments, Bellvue, Wash) and found that it provides an equal view of angle structures. The double-mirror gonioscopic lens facilitates the intraoperative visualization of the anterior chamber angle and should allow surgeons who treat glaucoma to more accurately and safely perform angle-related procedures. PMID- 9338689 TI - Stereoscopic ophthalmic microendoscope system. AB - Technical characteristics of a new stereoscopic ophthalmic microendoscope system developed for the direct observation of intraocular structures are described herein. Our system includes a stereoscopic visualization mechanism and an image superimposing system that projects the endoscopic image into the eyepiece of the surgical microscope, allowing the surgeon to monitor 3-dimensional endoscopic and microscopic images simply by looking into the eyepiece of the surgical microscope. Stereoscopic visualization of the intraocular structure facilitates fine manipulation of the endoscope probe just above the retina or ciliary body, providing more reliable and safer control of the probe within the eye. Preliminary clinical experience has demonstrated the usefulness of this new system. PMID- 9338690 TI - Oculosporidiosis. PMID- 9338691 TI - Multiple evanescent white dot syndrome in a patient with Best disease. PMID- 9338692 TI - Laser therapy for retinopathy of prematurity. PMID- 9338693 TI - Baseball hitting and the Pulfrich phenomenon: could it be due to traumatic optic neuropathy? PMID- 9338720 TI - Endoscopic substances for the treatment of vesicoureteral reflux. AB - The endoscopic treatment of reflux, like that of urinary incontinence, is effective. Several materials and endoscopic delivery systems are currently under evaluation for the treatment of reflux and incontinence. These include silicone microimplants, glass particles, collagen, dextranomer microspheres, a detachable balloon system, chondrocytes, and muscle cells. Although the endoscopic treatment is rendered in a similar fashion for both urinary incontinence and reflux, the acceptable safety and efficacy parameters of the bulking agents may differ, depending on the condition treated and the age of the patient. A review of the endoscopic bulking substances and systems currently available is presented. PMID- 9338721 TI - Intralesional administration of biological response modifiers in the treatment of localized cancer of the prostate: a feasibility study. AB - OBJECTIVES: The primary aims of this pilot study were to establish the feasibility of intraprostatic administration of biological response modifiers (BRMs) and to investigate the toxicity and side effects of recombinant interferon (IFN)-alpha-2b injected into prostate glands harboring cancer. A secondary goal was to perform a preliminary assessment of the antitumor effect of this treatment. METHODS: Nine patients with histologically documented carcinoma of the prostate participated in the study. IFN was administered weekly for 5 weeks, under transrectal ultrasound visualization, with a modified gun that permitted the controlled injection of small volumes initially into the area of the tumor and later into the whole gland. Total doses of IFN ranged between 15 and 100 MU. RESULTS: The procedure resulted in minor local discomfort, comparable to a prostatic biopsy. Side effects from the drug were those anticipated from most BRMs (chills, fever, malaise, headache, fatigue), and in every case they were minor and self-limiting to several hours. Local adverse events were limited to gross hematuria (2 patients [22%]) and hematospermia (1 patient [11%]) and resolved spontaneously within 2 weeks. Antitumor activity, a secondary goal of the study, was noted in 3 (33%) patients with limited follow-up (mean 22.5 months). CONCLUSIONS: The results of this pilot study indicate that the intraprostatic administration of IFN-alpha-2b can be readily accomplished by the method described here and is associated with minor, self-limited toxicity. With the regimen and doses used, IFN demonstrated modest antineoplastic activity. Modifications of the schedule, routes, and amounts administered may result in enhanced therapeutic value. PMID- 9338722 TI - Ethnic and geographic diversity of stone disease. AB - OBJECTIVES: To ascertain diversity or similarity in stone prevention and problems among different countries around the world. METHODS: Urolithiasis research groups from 10 countries completed a questionnaire. RESULTS: Cost of extracorporeal shock wave lithotripsy (ESWL) was considerably greater than that of drugs in four countries, and equivalent in remaining countries. Stone composition was similar among different countries. Certain urinary risk factors were associated with particular countries, probably from dietary indiscretions. ESWL was used in the majority of patients and open surgery in a minority of patients, except in one country. Medical diagnostic evaluation was used in the majority of patients except in one country. Drug treatment was nonselective, and provided to a minority of recurrent stone-formers. CONCLUSIONS: There is considerable similarity in stone presentations and problems throughout the world. The diversity is likely to be due to nutritional-environmental and socio-political economic factors. PMID- 9338724 TI - Urolithiasis associated with the protease inhibitor indinavir. AB - OBJECTIVES: To report the association between the protease inhibitor indinavir and the development of urolithiasis. METHODS: Case reports of three adult patients infected with the human immunodeficiency virus who developed surgical renal stones while being treated with indinavir are presented. RESULTS: Of the 3 patients requiring surgical intervention, stone analyses were available in 2. One stone revealed an inner core of an unidentifiable crystal surrounded by calcium oxalate, and another was found to have indinavir components as determined by thin layer chromatography and gas chromatography-mass spectrometry. Metabolic evaluation of all 3 patients identified significant hypocitraturia as an isolated finding. CONCLUSIONS: The widely used protease inhibitor indinavir is associated with the development of urolithiasis and may act as a nidus for heterogeneous nucleation leading to the development of mixed urinary stones. Surgical intervention may be necessary in some cases. Underlying metabolic abnormalities may contribute to the increased incidence of stone formation. Urologists and other health care providers should be aware of this association, as combined medical and surgical intervention may be necessary. PMID- 9338723 TI - Protease inhibitor-induced urolithiasis. AB - OBJECTIVES: To describe protease inhibitor-induced urinary stone disease in patients with human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS) who are taking indinavir sulfate (Crixivan), a protease inhibitor, for the treatment of AIDS. METHODS: Patients with HIV/AIDS and symptomatic renal colic temporally related to the initiation of indinavir sulfate therapy were prospectively identified. Seven patients (mean age 42 years; all men) with HIV and renal colic who were taking indinavir were identified. Retrospective chart reviews and patient interviews were performed. RESULTS: Indinavir therapy averaged 5.7 months prior to presentation with renal colic. All patients had microscopic hematuria. One patient presented with acute azotemia from bilateral urinary obstruction. Six patients had no history of urinary stones prior to initiating indinavir. The median number of symptomatic urinary stone episodes after initiating indinavir was two stones per patient. All patients had moderate- to high-grade urinary obstruction from radiolucent calculi. Abdominal computed tomography (CT) demonstrated hydronephrosis without urinary calcifications. Three patients spontaneously passed stones and 4 required intervention. Yellow debris and/or brown matrix-like material was seen endoscopically. Stone analysis revealed pure protease inhibitor. Six patients (86%) eventually discontinued protease inhibitor therapy. CONCLUSIONS: Protease inhibitor-induced urinary stones are radiolucent and can cause high-grade ureteral obstruction. Protease inhibitor-induced urinary stones were not identified on unenhanced abdominal CT scans. The radiolucent gelatinous nature of such stones makes lithotripsy a poor choice of treatment. Ureteral stenting may allow spontaneous stone passage if symptomatic obstruction occurs. Urologists may encounter a greater number of patients with symptomatic protease inhibitor induced urinary calculi as these medications become more popular. PMID- 9338725 TI - Spiral computed tomography for staghorn calculi. AB - OBJECTIVES: To assess the utility of spiral computed tomography (CT) with three dimensional reconstruction for preoperative planning of percutaneous nephrostolithotomy in patients with complex branched calculi (full staghorns). METHODS: Patients with complex branched stones were imaged with spiral CT with three-dimensional reconstruction. These images were compared with standard imaging modalities, including excretory urography and plain radiographs, for planning percutaneous access for nephrostolithotomy. The utility of the scan was evaluated. RESULTS: Ten patients with branched calculi were studied. Anatomic abnormalities were present in 5 patients. Excellent three-dimensional images were obtained in all patients without any complications related to the study. In 1 patient with multiple calculi in a horseshoe kidney, the three-dimensional image indicated a branched stone. The spiral CT scan was not helpful in directing percutaneous access in any patient. In a single patient, residual fragments noted during nephrostolithotomy were located by reference to the spiral CT scan. CONCLUSIONS: Spiral CT scans with three-dimensional reconstruction provide three dimensional imaging of branched renal calculi. This modality provides minimal additional information over that obtained from standard radiographic studies for guiding nephrostolithotomy and cannot be recommended as a routine preoperative study. It was helpful in 1 patient to locate a residual fragment. PMID- 9338726 TI - Bladder cancer in renal transplant recipients. AB - OBJECTIVES: To determine if there is an increased rate of occurrence of bladder cancer in renal transplant recipients. METHODS: We reviewed the records of 3130 consecutive renal transplant recipients at the University of Wisconsin Hospital from 1980 to 1994. We then compared the rate of occurrence of bladder cancer in this population with that of the general population of Wisconsin. Using an age specific hazard model based on the rate of bladder cancer in the general population of Wisconsin, we predicted the expected number of bladder cancer cases in this renal transplant population. RESULTS: Using this model, one would expect 1.81 cases of bladder cancer in the renal transplant data set, as opposed to the observed 6, resulting in a relative risk of 3.31 of developing bladder cancer as a result of renal transplantation. CONCLUSIONS: There is a higher incidence of bladder cancer in renal transplant recipients. Therefore, despite a higher incidence of hematuria in this population, each noninfected patient with microscopic (or gross) hematuria should undergo a careful urologic evaluation. PMID- 9338727 TI - Intravesical immunoprophylaxis in recurrent superficial bladder cancer (Stage T1): multicenter trial comparing bacille Calmette-Guerin and interferon-alpha. AB - OBJECTIVES: To estimate and compare recurrence rates, index of recurrence, and disease-free interval in patients with superficial recurrent bladder cancer receiving bacille Calmette-Guerin (BCG) or interferon (IFN) for immunoprophylaxis. METHODS: One hundred twenty-two patients with recurrent superficial Stage pT1, grade 1 to 3 tumors were enrolled in a randomized, prospective, multicenter trial with two treatment arms of endovesical immunoprophylaxis: 150 mg of BCG versus 54 MU of recombinant IFN-alpha-2a. Administration was weekly during the first month, biweekly for 2 months, and monthly for 9 months. Both groups were similar with regard to tumor stage, grade, size, and number. RESULTS: Sixty-one patients were evaluable in the BCG group and 49 in the IFN group. Tumors recurred in 34 (69.4%) of 49 patients in the IFN group (890 months of follow-up) and in 24 (39.3%) of 61 in the BCG group (1272 months of follow-up). The total number of recurrences (28 for BCG, 47 for IFN), disease-free interval (mean 19.3 months for BCG, 15.3 months for IFN), and index of recurrence (2.2 for BCG, 5.5 for IFN) were statistically significant (P = 0.001) in favor of BCG. Progression to invasive carcinoma was similar in both study arms. Neither systemic nor local side effects were seen in the IFN group. However, the previously reported toxicity of BCG was confirmed. CONCLUSIONS: According to our trial, BCG remains the most efficacious agent for immunoprophylaxis of recurrent superficial bladder tumors. PMID- 9338728 TI - Incidence of urethral involvement in female bladder cancer: an anatomic pathologic study. AB - OBJECTIVES: To evaluate risk factors for urethral involvement in female patients undergoing cystectomy for bladder cancer and to define potential candidates for orthotopic continent urinary diversion. METHODS: From 1990 to 1996, 43 female patients underwent cystectomy for primary bladder malignancy. Bladder mapping studies were performed with special attention to tumor location, multifocality, pathologic stage and grade, presence of carcinoma in situ (CIS), and the relationship of these factors to urethral involvement. RESULTS: Of the 43 patients evaluated, 7 (16.3%) had urethral involvement by tumor. Two patterns of urethral invasion were identified: 5 patients had tumors in the vascular or lymphatic channels of the periurethral tissue, and 2 had tumors in the mucosa or submucosa. Three of the 12 patients with tumors located at the trigone and 4 of the 26 with CIS of the bladder had tumor in the urethra. Three of the 5 patients with vaginal involvement had tumor in the urethra. Only vaginal involvement was significantly associated with urethral involvement (P = 0.02). CONCLUSIONS: Vaginal involvement was the only preoperative factor that was associated with presence of tumor in the urethra. Five patients with urethral involvement had submucosal tumors without concomitant CIS of the urethra. Before the performance of orthotopic urinary diversions, intraoperative full-thickness bladder neck biopsies are needed to accurately evaluate the female urethra. PMID- 9338729 TI - Questionnaire survey of urologists and primary care physicians' diagnostic and treatment practices for prostatitis. AB - OBJECTIVES: To establish diagnostic and treatment practices for chronic prostatitis by survey of urologists in Wisconsin and primary care providers in Dane County, Wisconsin. METHODS: All Wisconsin urologists (n = 135) and primary care providers in Dane County, Wisconsin (n = 365) were surveyed by mail with a 10-item questionnaire used to establish diagnostic and treatment practices for prostatitis. RESULTS: Seventy-eight percent of primary caregivers consider prostatitis to be bacterial in nature, whereas 59% of urologists consider it to be noninfectious. Fewer than 50% of primary care providers consider pain to be other than perineal in the diagnosis. Fewer than 50% of urologists or primary caregivers evaluate expressed prostatic secretions and few primary care providers (11%) use nonantibiotic therapy. CONCLUSIONS: The diagnostic and treatment practices for prostatitis do not follow standard textbook algorithms. PMID- 9338730 TI - Correlation of presumed circle area ratio with infravesical obstruction in men with lower urinary tract symptoms. AB - OBJECTIVES: To examine the predictive value of ultrasonic measurements obtained by transrectal ultrasonography for infravesical obstruction as evaluated by pressure flow studies. METHODS: In 85 men with moderate to severe lower urinary tract symptoms, ultrasonic measurements including prostatic volume, transition zone volume, transition zone index (transition zone volume/prostatic volume), and presumed circle area ratio (PCAR) were compared with urodynamic parameters obtained by pressure flow studies. RESULTS: There were significant interrelationships between these ultrasonic measurements, which were all significantly greater in the obstructed patients than in the unobstructed patients. A simple regression analysis demonstrated that prostatic volume (r = 0.362, P < 0.001), transition zone volume (r = 0.373, P < 0.0005), transition zone index (r = 0.331, P < 0.005), and PCAR (r = 0.487, P < 0.0001) correlated significantly with the Abrams-Griffiths number. More importantly, a multiple regression analysis demonstrated PCAR to be the only independent determinant of the Abrams-Griffiths number. A receiver operator characteristics curve analysis showed that 0.8 was the most suitable cutoff value of PCAR for the prediction of infravesical obstruction with a diagnostic accuracy of 76.5%. CONCLUSIONS: PCAR is useful as a transrectal ultrasonic measurement in assessing the severity of infravesical obstruction in men with lower urinary tract symptoms. PMID- 9338731 TI - Serial prostate-specific antigen measurements in men with clinically benign prostatic hyperplasia during a 12-month placebo-controlled study with terazosin. HYCAT Investigator Group. Hytrin Community Assessment Trial. AB - OBJECTIVES: To prospectively analyze whether the treatment of men with clinically benign prostatic hyperplasia (BPH) with alpha blocking agents affects the serum prostate-specific antigen (PSA) levels, and to determine the magnitude of such effect. METHODS: Serial PSA measurements were performed using the Abbott IMx assay over 1 year in 134 men over the age of 55 years participating in the Hytrin Community Assessment Trial (HYCAT). HYCAT is a 1-year, randomized, placebo controlled, double-blinded study of the alpha1-adrenergic antagonist terazosin. All men had lower urinary tract symptoms and a clinical diagnosis of BPH with an American Urological Association (AUA) symptom index of 13 points or more, an AUA bother score of 8 points or more, and a peak urinary flow rate of less than 15 mL/s. PSA was measured at baseline and at 8, 26, 39, and 52 (end of study) weeks. RESULTS: Baseline serum PSA levels weakly correlated with patients' age at study entry, and modestly with residual urine (positive correlation) and peak flow rate (negative correlation), although none of the levels were statistically significant. Changes of serum PSA during the course of the study did not correlate with either one of the symptom severity or bother assessment tools, residual urine, or peak flow rate. Mean PSA increased from a baseline of 2.5+/ 0.22 ng/mL (mean+/-SE) by 0.5+/-0.11 ng/mL in the placebo-, and from 2.7+/-0.23 ng/mL by 0.3+/-0.11 ng/mL in the terazosin-treated patients (P = 0.36 by ANOVA). There were no differences in the changes in serum PSA when patients were stratified by decade of life according to the age-specific PSA reference ranges, or by the final dose of terazosin (2, 5, or 10 mg daily). CONCLUSIONS: The treatment of men with lower urinary tract symptoms and clinical BPH with the alpha1-adrenergic antagonist terazosin does not affect serum PSA concentration, and thus does not confound longitudinal monitoring of serum PSA levels in patients at risk for prostate carcinoma. PMID- 9338732 TI - The sextant protocol for ultrasound-guided core biopsies of the prostate underestimates the presence of cancer. AB - OBJECTIVES: The aim of this prospective study was to evaluate the sensitivity of the sextant biopsy protocol compared with a more extensive procedure for the detection of prostate cancer and to define a biopsy model with the minimal number of biopsies necessary to maintain diagnostic accuracy. METHODS: A total of 512 consecutive patients with suspected prostate cancer were examined with transrectal ultrasound (TRUS) and underwent TRUS-guided core biopsy. All patients had 8 or 10 standardized biopsy samples taken, with the number depending on the size of the gland. Additional biopsy samples were taken from hypoechoic or hyperechoic lesions located outside the predetermined location for the standardized biopsies (ie, target biopsies). The sensitivity of the detection of cancer for different combinations of biopsy samples was analyzed and compared with that of our model with 8 to 10 biopsies. RESULTS: In all, 276 cancers were detected, of which 88 (32%) had an isoechoic appearance. Sensitivity was 59% for focal lesions detected by TRUS, 85% to 97% for different combinations of systematic biopsy samples, and 93% to 98% for a combination of systematic and target biopsy samples. The sensitivity for the standard sextant protocol was 85%. By adding target biopsies, the sensitivity increased to 93%. CONCLUSIONS: The standard sextant protocol leaves 15% of cancers undetected compared with results obtained from a more extensive biopsy procedure. By combining systematic and target sampling, the sensitivity increases; however, a major concern is that the clinical importance of cancers detected by multiple biopsies needs to be evaluated. PMID- 9338733 TI - Cross-reactivity of ten anti-prostate-specific antigen monoclonal antibodies with human glandular kallikrein. AB - OBJECTIVES: Prostate-specific antigen (PSA) is commonly used as a marker for prostate disease. Prostate epithelium expresses both PSA and human glandular kallikrein (hK2) proteins, which share 80% sequence homology. The immunologic cross-reactivity of these two proteins could potentially interfere with determination of PSA levels in diagnoses of prostate cancer. We set out to determine the extent of this cross-reactivity for a panel of 10 anti-PSA monoclonal antibodies (mAbs). METHODS: Enzyme-linked immunosorbent assay (ELISA), sandwich assays, and western transfer techniques were used to assess the PSA/hK2 cross-reactivity of the anti-PSA mAbs. RESULTS: We did not detect the hK2 protein with any of the 10 anti-PSA mAbs under western transfer conditions. In ELISA experiments, 8 of 10 mAbs exhibited hK2 cross-reactivity under certain conditions. However, no combination of mAbs tested in sandwich assays exhibited a signal in hK2 cross-reactivity experiments greater than 0.1% of the PSA signal. CONCLUSIONS: We have evaluated 10 anti-PSA mAbs and determined that despite the 80% homology between PSA and hK2 proteins, cross-reactivity with hK2 by these antibodies would not significantly affect the determination of PSA levels by means of sandwich assays. PMID- 9338735 TI - Orthotopic urinary diversion is a viable option in patients undergoing salvage cystoprostatectomy for recurrent prostate cancer after definitive radiation therapy. AB - OBJECTIVES: To evaluate whether orthotopic urinary diversion is a viable option for patients undergoing cystoprostatectomy for radio-recurrent prostate cancer (RRPC). METHODS: Between 1990 and 1996, we performed 34 salvage surgeries for RRPC, including 26 radical retropubic prostatectomies and 8 cystoprostatectomies. We determined the operative and postoperative complication rates and pathologic stage for the 8 patients undergoing cystoprostatectomy. RESULTS: Of the 8 patients in whom cystoprostatectomy was performed, 5 underwent ileal conduit diversion and 3 underwent orthotopic neobladder reconstruction. There were no intraoperative complications or perioperative mortalities. In the group with orthotopic neobladder, postoperative complications included pyelonephritis in 1 patient and prolonged ileus in another. In the group with ileal conduit, no short term complications occurred; 1 patient developed an incisional hernia on long term follow-up. All patients with neobladder reconstruction are continent during the day. One patient wears one pad at night. The other 2 are continent at night. CONCLUSIONS: Orthotopic urinary diversion is a valid option for selected patients with RRPC who require a cystoprostatectomy. This procedure can be performed with minimal complications, resulting in good continence and good quality of life. PMID- 9338734 TI - Early detection of recurrent prostate cancer with an ultrasensitive chemiluminescent prostate-specific antigen assay. AB - OBJECTIVES: Treatment failure after radical prostatectomy is most commonly heralded by an increase in serum prostate-specific antigen (PSA) to detectable levels. We evaluated the clinical utility of an ultrasensitive chemiluminescent PSA assay. METHODS: We evaluated the assay in banked sera obtained from 170 men after radical prostatectomy. Controls consisted of 142 females, 29 men who had undergone cystoprostatectomy without evidence of prostate cancer, and 25 men without evidence of recurrent disease at least 5 years after prostatectomy for organ-confined disease. Lead time to diagnosis of recurrence was based on comparisons with the IMx or Tandem E assays using a cutoff of 0.1 ng/mL (100 pg/mL). RESULTS: The biologic level of detection of this assay is 8 pg/mL. Serum PSA levels were undetectable in 82.4% of females, 86.2% of the cystoprostatectomy patients, and 96% of the radical prostatectomy controls. After radical prostatectomy, PSA levels were undetectable at last check in 104 of 168 (61.9%) men. In the 24 men with prostate cancer recurrence, the enhanced sensitivity of 8 pg/mL provided a mean lead time based on conservative calculations of 12.7 to 22.5 months over conventional assays. Thirty-four of the 41 men with detectable PSA levels and no evidence of disease recurrence had PSA levels of 30 pg/mL or less. CONCLUSIONS: PSA levels are undetectable in most men who do not have recurrence of disease after radical prostatectomy. Low but detectable serum PSA levels less than or equal to 30 pg/mL can be produced by nonmalignant sources of PSA. PSA assays with enhanced sensitivity can detect recurrent prostate cancer with significant lead time over conventional assays. PMID- 9338736 TI - Feasibility of outpatient percutaneous bladder neck suspension under local anesthesia. AB - OBJECTIVES: To demonstrate the feasibility of outpatient percutaneous bladder neck suspension (BNS) under local anesthesia to treat stress urinary incontinence (SUI) in females. METHODS: Since October 1994, 40 women with SUI (mean age 59.6+/ 12.0 years) underwent outpatient percutaneous BNS with "Z" suture anchoring of the anterior vaginal wall and pubocervical fascia. The suspension sutures were secured to percutaneously placed bone anchors. The procedure was performed under local anesthesia. Pain during surgery was evaluated with an analogue scale graded from 0 (no pain) to 5 (severe pain). RESULTS: In 98% of the BNS, the procedure was successfully performed with the patient under local anesthesia. Conversion to general anesthesia was necessary for only 1 patient due to knee pain in the lithotomy position. No major complications were observed. Patients rated their perioperative pain as 1.0+/-0.6 (that is, minimal pain). Patients required pain medications for a mean of 2.4+/-1.3 days postoperatively. Mean duration of recovery (defined by a return to normal activities) was 2.2+/-1.0 weeks; 92% of the patients were continent postoperatively, and no recurrence of urethral hypermobility was observed. CONCLUSIONS: Our results demonstrate the feasibility of outpatient percutaneous BNS performed in patients under local anesthesia without significant morbidity. Continued follow-up will be necessary to determine the long-term efficacy of this procedure. PMID- 9338737 TI - Active surveillance after orchiectomy for nonseminomatous testicular germ cell tumors: late relapse may occur. AB - OBJECTIVES: To review the outcome of men with Stage I nonseminomatous germ cell tumors managed with a policy of active surveillance following orchiectomy. METHODS: The clinical records of all men with Stage I nonseminomatous germ cell tumors seen at Royal Prince Alfred Hospital, Australia between 1982 and 1995 were reviewed. Data were obtained concerning the histologic type of tumor, levels of serum tumor markers, relapse and subsequent treatment, and survival. RESULTS: Seventy-seven patients were entered into the active surveillance protocol between 1982 and 1995. With a minimum follow-up of 2 years, 27 (35%) have relapsed, with a median time to relapse of 5 months. Two late relapses occurred at 37 and 57 months after diagnosis. Relapses occurred most commonly in the retroperitoneal lymph nodes, with the lungs the second most common site. Following treatment with chemotherapy and surgery, all patients achieved complete remission, with 1 patient subsequently relapsing and ultimately dying of progressive tumor. One other patient died of acute myeloid leukemia, thought to be secondary to chemotherapy. Overall, 75 patients (97%) remain alive and free of disease. CONCLUSIONS: Active surveillance is a safe and effective approach to the management of Stage I nonseminomatous germ cell tumors. Although most relapses occur within the first 2 years, late relapses may occur. PMID- 9338738 TI - Quality of life after partial penectomy for penile carcinoma. AB - OBJECTIVES: To investigate the impact of partial penectomy on the quality of life of patients with carcinoma of the penis. METHODS: Fourteen patients who had undergone partial penectomy for penile cancer were studied. Their median age was 50.5 years and the median time of follow-up was 11.5 months. The quality of life was evaluated in three dimensions: social adjustment, sexuality, and emotional state. The patients underwent a semistructured interview and were asked to complete the Overall Sexual Functioning Questionnaire, the Social Problem Questionnaire, the General Health Questionnaire, and the Hospital Anxiety and Depression Scale. RESULTS: In 9 (64%) patients, the overall sexual function was normal or slightly decreased. Only 2 (14%) men had precarious or absent sexual function. The masculine self-image and the relationship with their partners remained practically unchanged in all the patients. Sexual interest and satisfaction remained normal or slightly reduced in 9 and 12 patients, respectively. The frequency of sexual intercourse was unchanged or slightly decreased in 9 patients. Three patients had no sexual intercourse after surgery. No significant levels of anxiety and depression were found. Within the areas of living conditions, family life, and interactions with other people, all the patients remained as they were before the surgery. CONCLUSIONS: Patients who undergo partial penectomy for penile cancer can maintain the quality of life (in social, psychological, and sexual terms) at levels similar to those that existed in the period before surgery. PMID- 9338739 TI - Modified pudendal-thigh flap for correction of penoscrotal transposition. AB - OBJECTIVES: In patients with penoscrotal transposition, an occasional postoperative problem has been a deficiency of skin on the proximal penile shaft that results in penoscrotal fusion and tethering. METHODS: We describe a new operation using a modified neurovascular pudendal-thigh flap for correction of incomplete penoscrotal transposition. RESULTS: This procedure has been used in 6 children, and an excellent cosmetic and functional result has been achieved in each patient. CONCLUSIONS: The flaps provide a reliable blood supply, maintain normal innervation, and correct the problem of postoperative penoscrotal fusion and tethering. This technique preserves sufficient penile skin for a tension-free second-stage urethroplasty. PMID- 9338740 TI - Magnetic resonance imaging in the evaluation of ureteropelvic junction obstructed kidney. PMID- 9338741 TI - Botryoid rhabdomyosarcoma of the bladder. PMID- 9338742 TI - Anuric renal failure from massive bilateral renal hematoma following extracorporeal shock wave lithotripsy. AB - We report a case of large bilateral perirenal and intrarenal hematoma formation leading to prolonged anuria in the absence of obstruction following extracorporeal shock wave lithotripsy. With conservative management, including the need for hemodialysis support, renal function gradually returned. A mechanism for this patient's anuric renal failure is postulated and caution is advised when considering simultaneous bilateral extracorporeal shock wave lithotripsy in patients with a potential risk of bleeding. PMID- 9338743 TI - Pseudotumor of the bladder: a late complication of inguinal herniorrhaphy. AB - Damage to the bladder during inguinal hernia repair is possible especially if the bladder or a bladder diverticulum is involved in the hernia sac. Unrecognized injury to the bladder can lead to late complications. We report a case of pseudotumor in a bladder diverticulum due to long-term retention of a misplaced suture. The literature on bladder injury after inguinal herniorrhaphy and on pseudotumor formation is briefly reviewed. It is important to be aware of a history of inguinal surgery and to obtain definitive histologic evidence of malignancy prior to making the diagnosis of bladder carcinoma. This will avoid unnecessary radical surgery, chemotherapy, or radiation therapy. PMID- 9338746 TI - Female outlet obstruction after repeated collagen injections. AB - An elderly woman who presented with severe irritative lower urinary tract symptoms after three consecutive collagen injections was diagnosed as being obstructed. The diagnosis was based on urodynamic, cystoscopic, and radiographic findings that disclosed numerous collagen deposits obstructing the bladder outlet and the proximal urethra. A transurethral resection of the collagen was performed with improvement in symptoms and flow pattern. Although irritative lower urinary tract symptoms after collagen injection could be related to local inflammatory changes, persistence and eventually worsening of these symptoms should alert to the presence of iatrogenic bladder outlet obstruction. PMID- 9338745 TI - Stone formation in the prostatic urethra after cryotherapy for prostate cancer. AB - We report on 3 cases of stone formation in the prostatic urethra after cryosurgical ablation of the prostate. This complication occurred late in the course, many months after the normal postoperative healing process apparently was finished and patients enjoyed normal voiding. Transrectal ultrasound proved to be useful in making the diagnosis. Treatment included lithotripsy and cold resection of residual dead tissue. PMID- 9338744 TI - Bleeding ileal conduit stomal varices: diagnosis and management using transjugular transhepatic angiography and embolization. AB - An uncommon complication of ileal conduit urinary diversion is bleeding varices at the stoma site. Variceal formation is a complication of portal hypertension, which is most commonly due to intrinsic liver disease. Problematic recurrent bleeding is usually managed locally or by portosystemic shunt. We report a case of recurrent, massive ileal conduit variceal hemorrhage in a patient without a significantly elevated portosystemic gradient. Therefore, this patient was not a candidate for a shunt procedure. Using a transjugular transhepatic approach to the portal vein, the varices were embolized to stasis without any complications. The patient has subsequently experienced no further bleeding episodes. PMID- 9338747 TI - Penile calciphylaxis. AB - Calciphylaxis is a condition of cutaneous necrosis secondary to small- and medium sized vessel calcification that may progress rapidly and is often fatal. Patients with end-stage renal disease and hyperparathyroidism are almost exclusively at risk. Only 1 case of penile involvement has been previously described. At our institution, a 56-year-old man with end-stage renal disease presented with penile calciphylaxis. The patient received a series of treatments including circumcision, partial penectomy, amputation of necrotic phalanges, and a subtotal parathyroidectomy after which the patient's parathyroid hormone level normalized and the disease progression abated. PMID- 9338748 TI - Reconstruction of ureteral defects with a tubular skin graft secondarily prefabricated utilizing omentum as a carrier: an experimental study. AB - OBJECTIVES: This study was conducted to evaluate reconstruction of the ureter in dogs after resection of a 7-cm-long midlength segment with a vascularized tubular skin graft secondarily prefabricated utilizing omentum as a carrier. METHODS: Nine female mongrel dogs underwent surgery in which omental surface capabilities were used to create a vascularized prefabricated skin graft. After 4 weeks, all 9 dogs underwent the second surgical procedure, which included the resection of a 7 cm-long segment from the ureter and an interureteral anastomosis of the omentocutaneous cylindrical tube. At postoperative week 10, just before the third surgical procedure, intravenous urography was performed to evaluate the continuity of the treated ureter. Nephroureterectomy was also performed immediately after intravenous urography to obtain specimens for histologic analysis of the aforementioned tubular anastomosis. RESULTS: The continuity of the ureteral defect was restored with the help of the omentocutaneous cylindrical tube. There was no narrowing throughout the ureter and along the omentocutaneous cylindrical tube. Only minimal dilation occurred at the neoskin tube and at the ipsilateral collecting system. Histopathologically, there were capillary protrusions entering the skin graft from the omental vasculature. The columnar epithelium of the cutaneous cylindrical tube was completely preserved, and transitional ingrowth was present at the proximal and distal ends of the tube. CONCLUSIONS: The surgical procedure resulted in successful reconstruction of ureteral continuity by the use of a prefabricated omentocutaneous cylindrical tube in dogs. PMID- 9338749 TI - Early experience with the use of gastric segment in lower urinary tract reconstruction in adult patient population. AB - OBJECTIVES: To review our experience with the potential use of stomach as a substitute to bowel in lower urinary tract reconstruction in adults. METHODS: Twenty-two adult patients underwent lower urinary tract reconstruction using stomach. Fourteen patients had augmentation cystoplasty, and in 8 patients a continent reservoir was constructed; mean follow-up was 9.8 months. In 6 patients, gastric tube was constructed and used as catheterizable stoma. RESULTS: Renal functions remained stable or improved in all patients. Two patients developed hypochloremic alkalosis. There was significant decrease in urinary pH. All patients were completely continent, with no problems in mucus production. There was no mortality or significant morbidity. CONCLUSIONS: Stomach offers a good alternative to ileum or colon for bladder reconstruction. Stomach has various unique advantages, such as less mucus production, acidic milieu in the urine, and protection against hyperchloremic acidosis. PMID- 9338750 TI - p53 and HER-2 alterations in renal cell carcinoma. AB - OBJECTIVES: To investigate protein expression and genetic aberrations of p53 and HER-2 in renal cell carcinoma (RCC) as well as possible association of p53 and HER-2 abnormalities with the tumor behavior of RCC. METHODS: Formalin-fixed and paraffin-embedded tissue blocks from 70 patients with RCC were studied. Protein expression of p53 and HER-2 was assessed immunohistochemically. Following deoxyribonucleic acid extraction from the paraffin sections using a microdissection technique, p53 gene mutations within exons 5 to 8 were examined by polymerase chain reaction (PCR) single-strand conformation polymorphism and HER-2 gene amplification by a differential PCR. RESULTS: Of the 70 RCCs, 16 (22.9%) showed protein expression of p53, with 7 (10%) demonstrating mutations within exons 5 to 7. Additionally, 28 RCCs (40%) displayed protein expression of HER-2, with 12 (17.1%) demonstrating gene amplification. All cases showing p53 mutations or HER-2 amplification represented protein expression of p53 or HER-2, respectively. No cases with negative immunostaining for p53 or HER-2 protein displayed positive results at gene level. Statistical analysis revealed a close relationship between p53 protein expression and the tumor grade, as well as a significant association of HER-2 protein expression and gene amplification with the tumor stage of RCC. CONCLUSIONS: Our observations suggest that the abnormalities of p53 and HER-2 may be involved in the pathogenesis of RCC and that other mechanisms leading to protein expression of p53 and HER-2 may coexist within a single tumor in addition to the genetic aberrations. PMID- 9338751 TI - Allelic loss of 8p sequences in prostatic intraepithelial neoplasia and carcinoma. AB - OBJECTIVES: Previous work has suggested that prostatic intraepithelial neoplasia (PIN) may be a premalignant lesion important in tumorigenesis of the prostate. However, to adequately test this hypothesis at the genetic level, it is necessary to determine whether lesions in close proximity demonstrate similar genetic alterations and, hence, whether an "evolutionary" relationship might exist between PIN and tumor in the same prostate. Therefore, the purpose of this study was to examine at least two PIN lesions per prostate (one adjacent to and another distant from malignant lesions in the same prostate) for similarities or differences in the types and frequencies of genetic alterations. METHODS: To accomplish this goal, DNA was extracted from microdissected PIN, tumor, and normal epithelial tissue samples from 48 radical prostatectomies and amplified using polymerase chain reaction techniques at highly polymorphic microsatellite repeat sequences at proximal (D8S87, 8p12) and distal (NEFL, 8p21) loci on the short arm of chromosome 8. PIN specimens were either adjacent to (within one high power microscopic field [HPF]) or distant from (separated by two or more HPFs) tumor specimens from the same patients. RESULTS: Similar fractional allelic loss frequencies were observed for informative tumor (10 [35%] of 29) and PIN (6 [21%] of 29) samples at the NEFL locus, but allelic loss at the D8S87 locus was observed only in tumors (8 [22%] of 36 informative samples). Moreover, allelic loss at the NEFL locus involved the same allele in 4 cases and different alleles in 3 cases. Interestingly, all 4 cases with the same allele loss were from adjacent PIN and tumor tissues, and all 3 with different allele loss were from distant PIN and tumor. CONCLUSIONS: These results suggest that PIN and invasive cancer share common genetic events (eg, deletion at the NEFL locus) along the same pathway of development in the prostrate. PMID- 9338752 TI - Hugh Cabot--1872-1945: genitourinary surgeon, futurist, and medical statesman. PMID- 9338753 TI - Predicting probability of prostate cancer. PMID- 9338754 TI - Assisted reproduction cost-effectiveness analysis. PMID- 9338755 TI - Marked obesity and laparoscopic bladder neck suspension. PMID- 9338756 TI - Trust and the bioethics industry. PMID- 9338757 TI - Vaccines at risk. PMID- 9338758 TI - French ministries in argument over release of asbestos report. PMID- 9338760 TI - Row over alternative medicine's status at NIH. PMID- 9338759 TI - Mentally disabled research subjects 'need protection'. PMID- 9338761 TI - Islanders contest report on nuclear risks. PMID- 9338762 TI - Brazil to sequence 'first plant pathogen'. PMID- 9338763 TI - Vaccine institute treads out a wary path. PMID- 9338764 TI - Business booms for guides to biology's moral maze. PMID- 9338765 TI - Russia warned: act now or regret it later. PMID- 9338766 TI - Germany's past still casts a long shadow. PMID- 9338767 TI - France reaps benefits and costs of going by the book. PMID- 9338768 TI - Japan's bioethics debate lags behind thinking in the West. PMID- 9338769 TI - UK takes pride in 'principled pragmatism'. PMID- 9338770 TI - Policing ethical codes in India proves tough. PMID- 9338771 TI - Pre-empting the arrival of a dark lord. PMID- 9338772 TI - Developmental neurobiology. Unscrambling a disabled brain. PMID- 9338773 TI - Genomic imprinting. Making sense or antisense? PMID- 9338774 TI - G proteins. The arginine finger strikes again. PMID- 9338775 TI - Alzheimer's disease. The ins and outs of amyloid-beta. PMID- 9338776 TI - Evolutionary biology. Insights from the echinoderms. PMID- 9338777 TI - Two sets of human-tropic pig retrovirus. PMID- 9338778 TI - Homeotic transformation in Drosophila. PMID- 9338779 TI - An intracellular protein that binds amyloid-beta peptide and mediates neurotoxicity in Alzheimer's disease. AB - Amyloid-beta is a neurotoxic peptide which is implicated in the pathogenesis of Alzheimer's disease. It binds an intracellular polypeptide known as ERAB, thought to be a hydroxysteroid dehydrogenase enzyme, which is expressed in normal tissues, but is overexpressed in neurons affected in Alzheimer's disease. ERAB immunoprecipitates with amyloid-beta, and when cell cultures are exposed to amyloid-beta, ERAB inside the cell is rapidly redistributed to the plasma membrane. The toxic effect of amyloid-beta on these cells is prevented by blocking ERAB and is enhanced by overexpression of ERAB. By interacting with intracellular amyloid-beta, ERAB may therefore contribute to the neuronal dysfunction associated with Alzheimer's disease. PMID- 9338780 TI - A synthetic peptide ligase. AB - The preparation of synthetic molecules showing the remarkable efficiencies characteristic of natural biopolymer catalysts remains a formidable challenge for chemical biology. Although significant advances have been made in the understanding of protein structure and function, the de novo construction of such systems remains elusive. Re-engineered natural enzymes and catalytic antibodies, possessing tailored binding pockets with appropriately positioned functional groups, have been successful in catalysing a number of chemical transformations, sometimes with impressive efficiencies. But efforts to produce wholly synthetic catalytic peptides have typically resulted in compounds with questionable structural stability, let alone reactivity. Here we describe a 33-residue synthetic peptide, based on the coiled-coil structural motif, which efficiently catalyses the condensation of two shorter peptide fragments with high sequence- and diastereoselectivity. Depending on the substrates used, we observe rate enhancements of tenfold to 4,100-fold over the background, with catalytic efficiencies in excess of 10(4). These results augur well for the rational design of functional peptides. PMID- 9338781 TI - Radical alterations in the roles of homeobox genes during echinoderm evolution. AB - Echinoderms possess one of the most highly derived body architectures of all metazoan phyla, with radial symmetry, a calcitic endoskeleton, and a water vascular system. How these dramatic morphological changes evolved has been the subject of extensive speculation and debate, but remains unresolved. Because echinoderms are closely related to chordates and postdate the protostome/deuterostome divergence, they must have evolved from bilaterally symmetrical ancestors. Here we report the expression domains in echinoderms of three important developmental regulatory genes (distal-less, engrailed and orthodenticle), all of which encode transcription factors that contain a homeodomain. Our findings show that the reorganization of body architecture involved extensive changes in the deployment and roles of homeobox genes. These changes include modifications in the symmetry of expression domains and the evolution of several new developmental roles, as well as the loss of roles conserved between arthropods and chordates. Some of these modifications seem to have evolved very early in the history of echinoderms, whereas others probably evolved during the subsequent diversification of adult and larval morphology. These results demonstrate the evolutionary lability of regulatory genes that are widely viewed as conservative. PMID- 9338782 TI - Serrate2 is disrupted in the mouse limb-development mutant syndactylism. AB - The mouse syndactylism (sm) mutation impairs some of the earliest aspects of limb development and leads to subsequent abnormalities in digit formation. In sm homozygotes, the apical ectodermal ridge (AER) is hyperplastic by embryonic day 10.5, leading to abnormal dorsoventral thickening of the limb bud, subsequent merging of the skeletal condensations that give rise to cartilage and bone in the digits, and eventual fusion of digits. The AER hyperplasia and its effect on early digital patterning distinguish sm from many other syndactylies that result from later failure of cell death in the interdigital areas. Here we use positional cloning to show that the gene mutated in sm mice encodes the putative Notch ligand Serrate. The results provide direct evidence that a Notch signalling pathway is involved in the earliest stages of limb-bud patterning and support the idea that an ancient genetic mechanism underlies both AER formation in vertebrates and wing-margin formation in flies. In addition to cloning the sm gene, we have mapped three modifiers of sm, for which we suggest possible candidate genes. PMID- 9338783 TI - Severe neuropathies in mice with targeted mutations in the ErbB3 receptor. AB - Neuregulins and their specific receptors, members of the ErbB family of tyrosine kinases, have been implicated in the control of growth and development of Schwann cells, specialized cells that wrap around nerve axons to provide electrical insulation. Here we use gene targeting to generate mice that lack ErbB3, a high affinity neuregulin receptor. Homozygous erbB3 mutant embryos lack Schwann-cell precursors and Schwann cells that accompany peripheral axons of sensory and motor neurons. The initial development of motor neurons and sensory neurons of dorsal root ganglia occurs as it should, but at later stages most motor neurons (79%) and sensory neurons in dorsal root ganglia (82%) undergo cell death in erbB3 mutant embryos. Degeneration of the peripheral nervous system in erbB3 mutant pups is thus much more severe than the cell death in mice that lack neurotrophins or neurotrophin receptors. We also show that ErbB3 functions in a cell-autonomous way during the development of Schwann cells, but not in the survival of sensory or motor neurons. Our results indicate that sensory and motor neurons require factors for their survival that are provided by developing Schwann cells. PMID- 9338784 TI - Scrambler and yotari disrupt the disabled gene and produce a reeler-like phenotype in mice. AB - Formation of the mammalian brain requires choreographed migration of neurons to generate highly ordered laminar structures such as those in the cortices of the forebrain and the cerebellum. These processes are severely disrupted by mutations in reelin which cause widespread misplacement of neurons and associated ataxia in reeler mice. Reelin is a large extracellular protein secreted by pioneer neurons that coordinates cell positioning during neurodevelopment. Two new autosomal recessive mouse mutations, scramble and yotari have been described that exhibit a phenotype identical to reeler. Here we report that scrambler and yotari arise from mutations in mdab1, a mouse gene related to the Drosophila gene disabled (dab). Both scrambler and yotari mice express mutated forms of mdab1 messenger RNA and little or no mDab1 protein. mDab1 is a phosphoprotein that appears to function as an intracellular adaptor in protein kinase pathways. Expression analysis indicates that mdab1 is expressed in neuronal populations exposed to Reelin. The similar phenotypes of reeler, scrambler, yotari and mdab1 null mice indicate that Reelin and mDab1 function as signalling molecules that regulate cell positioning in the developing brain. PMID- 9338785 TI - Neuronal position in the developing brain is regulated by mouse disabled-1. AB - During mammalian brain development, immature neurons migrate radially from the neuroectoderm to defined locations, giving rise to characteristic cell layers. Here we show that targeted disruption of the mouse disabled1 (mdab1) gene disturbs neuronal layering in the cerebral cortex, hippocampus and cerebellum. The gene encodes a cytoplasmic protein, mDab1 p80, which is expressed and tyrosine-phosphorylated in the developing nervous system. It is likely to be an adaptor protein, docking to others through its phosphotyrosine residues and protein-interacting domain. The mdab1 mutant phenotype is very similar to that of the reeler mouse. The product of the reeler gene, Reelin, is a secreted protein that has been proposed to act as an extracellular signpost for migrating neurons. Because mDab1 is expressed in wild-type cortical neurons, and Reelin expression is normal in mdab1 mutants, mDab1 may be part of a Reelin-regulated or parallel pathway that controls the final positioning of neurons. PMID- 9338786 TI - A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevents colitis. AB - Induction and maintenance of peripheral tolerance are important mechanisms to maintain the balance of the immune system. In addition to the deletion of T cells and their failure to respond in certain circumstances, active suppression mediated by T cells or T-cell factors has been proposed as a mechanism for maintaining peripheral tolerance. However, the inability to isolate and clone regulatory T cells involved in antigen-specific inhibition of immune responses has made it difficult to understand the mechanisms underlying such active suppression. Here we show that chronic activation of both human and murine CD4+ T cells in the presence of interleukin (IL)-10 gives rise to CD4+ T-cell clones with low proliferative capacity, producing high levels of IL-10, low levels of IL 2 and no IL-4. These antigen-specific T-cell clones suppress the proliferation of CD4+ T cells in response to antigen, and prevent colitis induced in SCID mice by pathogenic CD4+CD45RB(high) splenic T cells. Thus IL-10 drives the generation of a CD4+ T-cell subset, designated T regulatory cells 1 (Tr1), which suppresses antigen-specific immune responses and actively downregulates a pathological immune response in vivo. PMID- 9338787 TI - Lipopolysaccharide-binding protein is required to combat a murine gram-negative bacterial infection. AB - An invading pathogen must be held in check by the innate immune system until a specific immune response can be mounted. In the case of Gram-negative bacteria, the principal stimulator of the innate immune system is lipopolysaccharide (LPS), a component of the bacterial outer membrane. In vitro, LPS is bound by lipopolysaccharide-binding protein (LBP) and transferred to CD14--the LPS receptor on the macrophage surface--or to high-density lipoprotein (HDL) particles. Transfer to CD14 triggers an inflammatory response which is crucial for keeping an infection under control. Here we investigate how LBP functions in vivo by using LBP-deficient mice. Surprisingly, we find that LBP is not required in vivo for the clearance of LPS from the circulation, but is essential for the rapid induction of an inflammatory response by small amounts of LPS or Gram negative bacteria and for survival of an intraperitoneal Salmonella infection. PMID- 9338788 TI - Imprinted expression of the Igf2r gene depends on an intronic CpG island. AB - Gametic imprinting is a developmental process that induces parental-specific expression or repression of autosomal and X-chromosome-linked genes. The mouse Igf2r gene (encoding the receptor for insulin- like growth factor type-2) is imprinted and is expressed from the maternal allele after embryonic implantation. We previously proposed that methylation of region 2, a region rich in cytosine guanine doublets (a 'CpG island') in the second intron of Igf2r, is the imprinting signal that maintains expression of the maternal allele. Here we use mouse transgenes to test the role of region 2 and the influence of chromosome location on Igf2r imprinting. Yeast artificial chromosome transgenes successfully reproduced the imprinted methylation and expression pattern of the endogenous Igf2r gene; deletion of region 2 from these transgenes caused a loss of imprinting and restored biallelic Igf2r expression. These results define a primary role for region 2 and a negligible role for chromosomal location in Igf2r imprinting; they also show that methylation imprints can maintain allelic expression. Short transgenes containing only region 2 and yeast artificial chromosome transgenes with an inactive Igf2r promoter do not attract parental specific methylation. All transgenes showing paternal-specific repression of Igf2r produced an antisense RNA whose transcription was dependent on region 2. The production of an antisense RNA by the repressed parental allele is reminiscent of the imprinting of the Igf2/H19 gene pair and may indicate that expression competition could play a general role in imprinting. PMID- 9338789 TI - Activation of ATP P2X receptors elicits glutamate release from sensory neuron synapses. AB - Painful stimuli to the skin initiate action potentials in the peripheral terminals of dorsal root ganglion (DRG) neurons. These action potentials propagate to DRG central terminals in the dorsal horn of the spinal cord, evoking release of excitatory transmitters such as glutamate onto postsynaptic dorsal horn neurons. P2X receptors, a family of ligand-gated ion channels activated by the endogenous ligand ATP, are highly expressed by DRG neurons. Immunoreactivity to P2X receptors has been identified in the dorsal horn superficial laminae associated with nociceptive DRG central terminals, suggesting the presence of presynaptic P2X receptors. Here we have used a DRG-dorsal horn co-culture system to show that P2X receptors are localized at presynaptic sites on DRG neurons; that activation of these receptors results in increased frequency of spontaneous glutamate release; and that activation of P2X receptors at or near presynaptic DRG nerve terminals elicits action potentials that cause evoked glutamate release. Thus activation of P2X receptors at DRG central terminals can modify sensory signal throughput, and might even initiate sensory signals at central synapses without direct peripheral input. This putative central modulation and generation of sensory signals may be associated with physiological and pathological pain sensation, making presynaptic P2X receptors a possible target for pain therapy. PMID- 9338790 TI - Molecular basis of agonism and antagonism in the oestrogen receptor. AB - Oestrogens are involved in the growth, development and homeostasis of a number of tissues. The physiological effects of these steroids are mediated by a ligand inducible nuclear transcription factor, the oestrogen receptor (ER). Hormone binding to the ligand-binding domain (LBD) of the ER initiates a series of molecular events culminating in the activation or repression of target genes. Transcriptional regulation arises from the direct interaction of the ER with components of the cellular transcription machinery. Here we report the crystal structures of the LBD of ER in complex with the endogenous oestrogen, 17beta oestradiol, and the selective antagonist raloxifene, at resolutions of 3.1 and 2.6 A, respectively. The structures provide a molecular basis for the distinctive pharmacophore of the ER and its catholic binding properties. Agonist and antagonist bind at the same site within the core of the LBD but demonstrate different binding modes. In addition, each class of ligand induces a distinct conformation in the transactivation domain of the LBD, providing structural evidence of the mechanism of antagonism. PMID- 9338791 TI - Structure at 1.65 A of RhoA and its GTPase-activating protein in complex with a transition-state analogue. AB - Small G proteins of the Rho family, which includes Rho, Rac and Cdc42Hs, regulate phosphorylation pathways that control a range of biological functions including cytoskeleton formation and cell proliferation. They operate as molecular switches, cycling between the biologically active GTP-bound form and the inactive GDP-bound state. Their rate of hydrolysis of GTP to GDP by virtue of their intrinsic GTPase activity is slow, but can be accelerated by up to 10(5)-fold through interaction with rhoGAP, a GTPase-activating protein that stimulates Rho family proteins. As such, rhoGAP plays a crucial role in regulating Rho-mediated signalling pathways. Here we report the crystal structure of RhoA and rhoGAP complexed with the transition-state analogue GDP.AlF4- at 1.65 A resolution. There is a rotation of 20 degrees between the Rho and rhoGAP proteins in this complex when compared with the ground-state complex Cdc42Hs.GMPPNP/rhoGAP, in which Cdc42Hs is bound to the non-hydrolysable GTP analogue GMPPNP. Consequently, in the transition state complex but not in the ground state, the rhoGAP domain contributes a residue, Arg85(GAP) directly into the active site of the G protein. We propose that this residue acts to stabilize the transition state of the GTPase reaction. RhoGAP also appears to function by stabilizing several regions of RhoA that are important in signalling the hydrolysis of GTP. PMID- 9338822 TI - Difficult choices. PMID- 9338823 TI - Regionalization and injury prevention and control--a new dynamic or persistent lethargy? PMID- 9338824 TI - Should we be advocates for injury prevention? PMID- 9338825 TI - Should injury prevention programmes be targeted? PMID- 9338826 TI - Population strategies for prevention? If only it were that simple! PMID- 9338827 TI - Injury prevention programmes in primary care: a high risk group or a whole population approach? AB - OBJECTIVE: To examine the relationship between risk factors for childhood unintentional injury and injury outcome and to assess the feasibility of using risk factors to identify children at high risk of injury. SETTING: One general practice in Nottingham, UK. METHOD: A postal questionnaire survey to all parents of children registered with the practice (n = 771) to obtain data on risk and sociodemographic factors. All children still registered with the practice one year later were followed up for occurrence of a medically attended injury. RESULTS: The response rate was 78%. The injury rate over the follow up year was 246 injuries per 1000 children. Previous medically attended injury was associated with each of the injury outcomes (odds ratio (confidence interval) for all attendances, 2.33 (1.37 to 4.05); for accident and emergency attendances, 2.27 (1.15 to 4.4); and for primary health care team attendances, 2.58 (1.33 to 5.0)). Male sex was associated only with accident and emergency department attendance (odds ratio 2.13 (1.06 to 4.2)). Maternal age and previous injury were associated with a higher number of injuries in the subsequent year on univariate and multivariate analyses. The sensitivity and positive predictive value of the risk factors were low, except for previous injury and male sex. The number of children needing an injury prevention intervention to prevent one injury as identified by the risk factors was not significantly different from that required if a whole population approach were to be used. CONCLUSION: Primary care based injury prevention programmes, at present, should not be targeted at children identified as being at 'high risk' of injury. Nevertheless, a larger study using a wider cross section of the population is needed to address this issue further. PMID- 9338828 TI - Non-fatal asphyxiation and foreign body ingestion in children 0-14 years. AB - OBJECTIVES: To examine the frequency and nature of non-fatal asphyxiation and foreign body ingestion injuries among children in the state of Victoria, Australia, and to identify possible areas for prevention. METHODS: For children under 15 years, all Victorian public hospital admissions, July 1987 to June 1995, due to asphyxiation or 'foreign body entering through other orifice' (which includes ingestions), were reviewed. Emergency department presentations due to asphyxiation and foreign body ingestion provided information on circumstances of, and the type of foreign bodies involved in the injuries. RESULTS: The childhood average annual admission rate for asphyxiation was 15.1 per 100,000. Food related asphyxiation peaked in infants under 1 year, and declined to low levels by 3 years. The main foods involved were nuts, carrot, apple, and candy. The rate of non-food related asphyxiation was relatively constant to 3 years of age and then declined by 6 years. Mechanical suffocation was less common. The annual admission rate for 'foreign body entering through other orifice' was 31.7 per 100,000. These injuries peaked in 2-3 year olds then gradually declined. About 80% of these foreign body admissions were ingestions, with coins being the major object ingested. Admission rates for these causes remained constant over the eight years. Asphyxiation resulted in a higher proportion admitted and longer hospital stays. CONCLUSIONS: Prevention of suffocation and strangulation needs to focus on a safe sleeping environment and avoidance of ropes and cords, while foreign body asphyxiation and ingestion needs a focus on education of parents and child carers regarding age, appropriate food, risk of play with coins, and other small items. Legislation for toy small parts could be extended to those used by children up to the age of 5 years, and to other products marketed for children. Design changes and warning labels also have a place in prevention. PMID- 9338829 TI - Factors affecting the severity of motor vehicle traffic crashes involving young drivers in Ontario. AB - OBJECTIVES: To assess the factors affecting the severity of motor vehicles traffic crashes involving young drivers in Ontario. POPULATION: Ontario young drivers, aged 16 to 20, involved in traffic crashes resulting in injury, between 1 January 1988 and 31 December 1993, on public roads in Ontario. METHODS: Population based case-control study. Cases were fatal injury, major injury, and minor injury crashes involving young drivers. Controls were minimal injury crashes involving young drivers. Cases and controls were obtained retrospectively from the Canadian Traffic Accident Information Databank. Unconditional logistic regression was used for data analysis. RESULTS: Factors significantly increasing the risk of fatal injury crashes include: drinking and driving (odds ratio (OR) 2.3), impairment by alcohol (OR 4.8), exceeding speed limits (OR 2.8), not using seat belts (OR 4.7), full ejection from vehicle (OR 21.3), intersection without traffic control (OR 2.2), bridge or tunnel (OR 4.1), road with speed limit 70-90 km/hour (OR 5.6) or 100 km/hour (OR 5.4), bad weather (OR 1.6), head-on collision (OR 80.0), and overtaking (OR 1.9). Results of the same model applied to major and minor injury crashes demonstrated consistent but weaker associations with decreasing levels of crash severity. CONCLUSIONS: A casual relationship between crash severity and the risk factors listed above was proposed. Risk factors recommended for preventive intervention include: alcohol consumption, speeding, and use of seat belts. Head-on collisions are of primary concern. PMID- 9338830 TI - Fatal and non-fatal farm injuries to children and adolescents in the United States, 1990-3. AB - OBJECTIVE: Examine the current magnitude of the injury problem to children and adolescents on farms, and to compare these data to that from 1978-83. DATA SOURCES: US National Center for Health Statistics Mortality Multiple Cause of Death Tapes for the years 1991-3, and the US Consumer Product Safety Commission National Electronic Injury Surveillance System for data on emergency department visits for 1990-3. SUBJECTS: Children and adolescents 19 years and younger injured on farms. RESULTS: There were an average of 104 deaths per year due to injuries occurring on farms. The rate of 8.0 deaths per 100,000 child farm residents is 39% lower than in 1979-81. More of the deaths occurred in hospital than previously. There were an average of 22,288 emergency department treated injuries per year. The rate of 1717 injuries per 100,000 child farm residents is 10.7% higher than 1979-83. Males were injured more frequently than females. Tractors accounted for 20.9% of all injuries, followed by horses (8.4%), all terrain vehicles and minibikes (8.0%), and farm wagons (7.7%). CONCLUSIONS: Farm injuries continue to be a major problem to children living on farms. While improved medical care may have contributed to the reduction in mortality, the continued high rate of injuries warrants study of a variety of intervention strategies to reduce the injury toll. There is also a need for ongoing injury surveillance to provide accurate data on the farm injury problem. PMID- 9338831 TI - Injury mortality among children and teenagers in New Zealand compared with the United States of America. AB - OBJECTIVES: New Zealand (NZ) has an unenviable track record in childhood injury mortality. We sought to describe this burden and to compare it with the United States of America (USA), with a view to taking the first step in identifying potential areas in which NZ might benefit from injury control as practiced in the USA. METHODS: We identified all children and teenagers who had died of injury for the period 1984-93 from the NZ Health Information Service mortality data files. We compared their rates of injury with previously published rates for USA. RESULTS: The age specific rates follow a J shaped distribution, with high rates in the first year of life followed by a decline to the lowest rate, among 5-9 year olds, a marginally higher rate among 10-14 year olds, and a dramatic rise among those in the 15-19 age group. The specific causes of death vary considerably by age group. NZ's overall rate of child and adolescent injury is not substantially different from that of the USA, but marked differences are apparent when examining cause specific rates. CONCLUSIONS: In terms of the theoretical potential to reduce the total injury mortality rate, priority must be given to 15-19 year olds who account for 61% of all NZ injury deaths. Priorities for this age group are: motor vehicle traffic crashes (especially those involving occupants and motorcyclists), and suicide. Among the children, priorities are: pedestrian and occupant deaths, and drownings. Among infants, the priority is suffocation. PMID- 9338832 TI - Economic costs of motor vehicle crashes involving teenaged drivers in Kentucky, 1994. AB - OBJECTIVES: To analyze data from motor vehicle crashes (MVCs) involving teenaged drivers in Kentucky for 1994, and derive cost estimates of these crashes. METHODS: Crash data were obtained from the Kentucky Traffic Accident Facts 1994 Report and the Kentucky Accident Reporting System. The National Highway Traffic Safety Administration's Crash-Cost program was used to generate cost estimates for Kentucky data. RESULTS: Teenaged drivers had significantly higher MVC fatal and non-fatal injury rates than did adult drivers. The deaths rates were 43.6 and 19.0 per 100,000 for teens and adult drivers, respectively. Odds ratios (ORs) were calculated to estimate the relative risk for (1) involvement in an MVC, (2) fatal or incapacitating injury, and (3) fatal injury for teenaged compared with adult drivers. The crude ORs were statistically significant at each age. Cost estimates were calculated on a per person/vehicle basis. A single fatal injury was $642,700. A critical injury was $563,000. In general, unit costs rose with increasing levels of injury severity. For the total number of fatal injuries, costs exceeded $91 million. For non-fatal injuries and property damage only crashes, total costs were $318 million. Overall, the total cost estimate for MVCs involving teenaged drivers was nearly $410 million. CONCLUSIONS: Strategies aimed to reduce the number of MVCs attributed to teenaged drivers should reduce both the number and costs of crash related deaths and injuries. Graduated driver licensing (GDL) systems are one plausible approach toward achieving this goal. By recently enacting a GDL system in Kentucky, it is anticipated that many lives and dollars will be saved. PMID- 9338833 TI - Estimation of age specific incidence rates of childhood burns from a prevalence survey of burn scars. AB - OBJECTIVES: This paper describes two methods of estimating the age specific incidence rates of childhood burns from a prevalence survey of burn scars. METHODS: A prevalence survey of burn scars was carried out in 1992 on 15,742 Ghanaian children aged 5 years or less. Nine hundred and fifty five (6.1%) of these children had scars from burn, and for 630 (66%) of these children, additional information about the burn incident, including the child's age at the time of the burn, was obtained from the mother two to three months later. Thirty four per cent of mothers of children with burn scars were not interviewed due to absence, relocation, or inaccessibility. Age specific incidence rates of burns were estimated for eight age groups using two methods. In method I, the number of incident cases of burns for each age group were estimated from the burn scars by subtracting the estimated contribution of scars from burns that had occurred at earlier ages. In method II, the estimate was based on the mother's recall of the age of the child at the time of the burn. RESULTS: Slightly different results were obtained with the two methods, and problems were noted with both methods. CONCLUSION: We recommend the use of these methods for estimating age specific incidence rates from retrospective population surveys for health conditions which result in long term residual markers. PMID- 9338835 TI - Introduction to statistics--1. The confidence interval. PMID- 9338834 TI - Minerva, have you led us astray? PMID- 9338836 TI - Medically attended injuries among young children: observations in a suburban area. 1964. AB - A control study of the effect of public health education on the rate of accidental injuries among children under 7 years of age in a newly developing suburban area provided an opportunity to gain an insight into the nature and extent of the accident problem itself during a one year baseline period preceding the educational phase of the study. An accident was arbitrarily defined as any actual or presumed trauma following an incident for which direct medical or dental attention was obtained. Data were gathered through regular visits of data collectors to physicians, dentists, and hospitals in the area. An annual accident rate of 124 per 1000 children under 7 years of age was found. Two or more accident occurred to 10% of the children. The highest injury rate by age (179 per 1000 children) was found among 2 year old children, with the rate amount 2 year old boys 75% higher than any other age-sex group. The possible relationships of suburban living to the type and location of accidents by age and sex and to the variation in accidents by day of week are also presented. PMID- 9338837 TI - A controlled study of health education in accident prevention: the Rockland County Child Injury Project. 1966. AB - The Rockland County Child Injury Prevention Project was designed to test the effects of public health education for parents on the incidence of accidental injuries to children under 7 years of age in a controlled situation. The incidence of accidental injuries, defined as any actual or presumed trauma following an accident for which medical or dental attention was obtained, was determined in a study population before, during, and after exposure to the educational program and in a comparable control group during a corresponding period of time. The study group was exposed to an intensive education program involving neighborhood discussion groups conducted by lay and professional leaders, meetings with organized groups, and a monthly newsletter. The study population was organized into small neighborhood units of about 24 homes within the suburban housing developments. To measure the effect of the education program, accident rates were computed for each three month reporting period, projected on an annual basis. No consistent differences were discernible in the trends in the accident rates between the study and control groups during the three years of the project. On the contrary, the curve for the study group crossed that of the control group on no less than five occasions. There was a sharp decline in accidents during the second half of the education phase. Any conclusions regarding a positive effect of the education program proved untenable in the light of the sharp and disproportionate increase in the accident rate in the study group during the six month phase after education had stopped. After what appeared to be a contrary trend earlier, developments during this period demonstrated vividly the need to continue reporting for a sufficient time in a controlled situation to be sure that fortuitous changes over short periods do not lead to unwarrented conclusion. The flow of reports from hospitals was more consistent than from physicians. This suggested that, in selected control studies, adequate rates for comparative purposes might have been obtained entirely from hospital records. PMID- 9338839 TI - Child Accident Prevention Trust in Northern Ireland. PMID- 9338838 TI - Pediatric injuries in an Arabian Gulf country. AB - OBJECTIVE: To determine the common types of injuries among children (0-14 years) in Al-Ain, United Arab Emirates (UAE). DESIGN: A retrospective descriptive hospital based study. SETTING: Al-Ain Medical District, Al-Ain Teaching Hospital, UAE. SUBJECTS: All patients aged 0-14 years seen at Al-Ain Teaching Hospital for injuries during 1994. RESULTS: The number of children with an injury who attended the emergency room was 16,518 (69.9% boys; 30.1% girls). Injury rates were higher among non-UAE nationals. The most frequent reason for hospital admission was poisoning (41%). In the age group < 5 years, the most common causes were falls, blunt trauma, and burns or scalds, while in the 5-9 year and in 10-14 year groups the most frequent cause was road traffic accidents (RTAs). Fights and sporting injuries were also seen frequently in children aged 10-14 years. CONCLUSION: Injury rates were higher in boys and RTAs mostly occurred in children over 10 years. The majority of cases (56%) occurred among non-UAE nationals, who are usually of lower socio-economic status. RECOMMENDATION: Injuries can be prevented by developing strategies to substantially increase the profile of health education to parents and children, by educating policy makers and health professionals, and by environmental modification, legislation, and enforcement. The UAE government can play an important part by establishing and supporting injury prevention programs with these goals. PMID- 9338840 TI - More safe communities programs in Scandinavia have been evaluated: repeating the results from Falkoping. PMID- 9338841 TI - The development of a multimedia teaching program for fiberoptic intubation. AB - Current training methods in fiberoptic intubation entail a trial and error process in which trainees acquire skills by practicing this technique in mannequins or patients. These training methods are not efficient and may expose patients to unnecessary instrumentation. An interactive software program is described which uses Director, a commercially available multimedia authoring tool, to (1) familiarize trainees with video images of the upper airway, (2) permit operator controlled progress through a normal fiberoptic intubation, (3) simultaneously display (side by side) two-dimensional or three-dimensional computer tomographic images with a fiberscope in place and the corresponding endoscopic video images, and (4) demonstrate some of the obstacles which occur in clinical practice (e.g. "white-out" and saliva). The intent of this package is to simulate fiberoptic intubation techniques as well as help one create a mental image of the path a fiberscope takes within the lumen of the upper airway. The potential for improving operator immersion (virtual reality) by using a more sophisticated input device is discussed. PMID- 9338842 TI - Modeling obstetric cardiovascular physiology on a full-scale patient simulator. AB - To our knowledge, this is the first attempt at adapting an existing cardiovascular model to simulate the hemodynamics of a particular patient population. Despite attempts to define the physiologic alterations in advance, we discovered there were critical parameters not completely defined in the literature. These were discovered through the iterative process of testing, comparing resulting vital signs with targets, and literature review. A list of the parameters that should be sought for future modeling efforts is provided (Table 3), but this list is by no means exhaustive. As further work is performed in this area, additional independent and essential parameters will be identified (pressure characteristics of valvular anomalies, for example). To define a physiology that is less well described in the literature, empirical alterations and best-guess estimates of parameter changes will be required with significantly more iterations. Finally, we have described only modeling of cardiovascular physiology, modeling the respiratory system will require a similar process. PMID- 9338844 TI - Neuromonitoring in defibrillation threshold testing. A comparison between EEG, near-infrared spectroscopy and jugular bulb oximetry. AB - OBJECTIVES: The aim was to study the physiological effects of induced ventricular fibrillation and subsequent circulatory arrest for defibrillation threshold testing on the brain using the EEG, jugular bulb oxymetry and near-infrared spectroscopy. METHODS: Thirteen patients undergoing surgery for implantable cardioverter-defibrillator implantation or replacement under general anesthesia were included. We continuously monitored the jugular bulb oxygen saturation (SjO2), regional oxygen saturation (rSO2) and the EEG. RESULTS: 59 episodes of circulatory arrest were studied. In all cases the rSO2 fell instantly while the EEG changed within 12 +/- 4 seconds after induction. The EEG indicated ischemic changes, ranging from occurrence of rhythmic delta activity to cessation of all electrical activity. On successful defibrillation the rSO2 increased to values in excess of pre-arrest levels and restored towards baseline; the SjO2 initially fell followed by a similar overshoot. Recovery times increased in proportion to arrest duration. CONCLUSION: Short lasting episodes of circulatory arrest have serious, but transient effects on brain function. The rSO2 is an effective non invasive tool for monitoring cerebral oxygenation during DFT-testing. PMID- 9338845 TI - Detection of dicrotic notch in arterial pressure signals. AB - OBJECTIVE: A novel algorithm to detect the dicrotic notch in arterial pressure signals is proposed. Its performance is evaluated using both aortic and radial artery pressure signals, and its robustness to variations in design parameters is investigated. METHODS: Most previously published dicrotic notch detection algorithms scan the arterial pressure waveform for the characteristic pressure change that is associated with the dicrotic notch. Aortic valves, however, are closed by the backwards motion of aortic blood volume. We developed an algorithm that uses arterial flow to detect the dicrotic notch in arterial pressure waveforms. Arterial flow is calculated from arterial pressure using simulation results with a three-element windkessel model. Aortic valve closure is detected after the systolic upstroke and at the minimum of the first negative dip in the calculated flow signal. RESULTS: In 7 dogs ejection times were derived from a calculated aortic flow signal and from simultaneously measured aortic flow probe data. A total of 86 beats was analyzed; the difference in ejection times was -0.6 +/- 5.4 ms (means +/- SD). The algorithm was further evaluated using 6 second epochs of radial artery pressure data measured in 50 patients. Model simulations were carried out using both a linear windkessel model and a pressure and age dependent nonlinear windkessel model. Visual inspection by an experienced clinician confirmed that the algorithm correctly identified the dicrotic notch in 98% (49 of 50) of the patients using the linear model, and 96% (48 of 50) of the patients using the nonlinear model. The position of the dicrotic notch appeared to be less sensitive to variations in algorithm's design parameters when a nonlinear windkessel model was used. CONCLUSIONS: The detection of the dicrotic notch in arterial pressure signals is facilitated by first calculating the arterial flow waveform from arterial pressure and a model of arterial afterload. The method is robust and reduces the problem of detecting a dubious point in a decreasing pressure signal to the detection of a well-defined minimum in a derived signal. PMID- 9338843 TI - Intrapartum reflectance pulse oximetry: effects of sensor location and fixation duration on oxygen saturation readings. AB - OBJECTIVE: To determine the effects of sensor location and suction fixation duration on measurements of intrapartum fetal oxygen saturation (SpO2) with a new reflectance pulse oximetry system. DESIGN: Fetal SpO2 values (n = 18) were determined in the first stage of labor before and after moving the sensor to another part of the fetal head. RESULTS: Mean fetal SpO2 values did not differ with sensor location (95% CI: -3.59 to 1.48). The duration of measurement period 1, before moving the sensor, was 104 +/- 44 (range 30-240) min. No time-dependent changes in SpO2 values were seen (r = 0.17). CONCLUSION: Suction is an effective and noninvasive method of securing the reflectance pulse oximetry sensor to the fetal head in the first stage of labor and does not interfere with reproducible SpO2 values over several hours. PMID- 9338846 TI - Functional anatomy of full-scale patient simulators. PMID- 9338847 TI - The Internet and electronic transmission of medical records. AB - OBJECTIVE: To review, from a legal perspective the potential for using the Internet for inter-institutional transfer of patient medical records. METHODS: Basic issues and recent legislation that relate to protection of both medical data, and those transferring that data over public network systems is reviewed. RESULTS: Many laws already in existence can be applied to Internet transmission, but questions of jurisdiction remain. Providing signatures on requests for information, which are in essence contracts, is a problem. Signatures must both prove the identity of the participants and provide for non-repudiation of the agreement. Cryptographic digital signatures appear secure and effective, but their use is difficult to implement. Simpler methods are fraught with risks, yet are more easily accomplished. The patient's rights of privacy must be balanced against the need for access by government, physician, or healthcare institutions to confidential information. In general, information holders must put forth reasonable efforts to keep information confidential. The development of acknowledged standards will provide guidance. Multiple laws provide some deterrence and hence some reassurance to healthcare institutions, for example, by criminalizing acts of electronic interception of patient records in transit. CONCLUSION: Some believe the expense of secure transfer of medical records by electronic means is a major obstacle; this is false: such transfers are now technologically quite easy. The greatest obstacle to electronic transfer of medical records at this point is the development of workable standards for signing agreements and protecting transmissions, but the perceived advantages will likely drive the necessary developments. PMID- 9338848 TI - Maker follows up on sevoflurane problem. PMID- 9338850 TI - Debate: The pulmonary artery catheter, is it safe? To use or not to use. Pro: A moratorium on PAC is unjustified. PMID- 9338851 TI - Debate: The pulmonary artery catheter, is it safe: To use or not to use. Con: Swan song for the Swan-Ganz? PMID- 9338852 TI - Do practitioner credentials help predict safety in anesthesia practice? PMID- 9338853 TI - The association between prenatal sella turcica morphology and notochordal remnants in the dorsum sellae. AB - The purpose of this study was to describe the location and morphology of notochordal remnants in the cranial base in normal and pathological conditions and to relate these findings to the morphological appearance of the sella turcica. Serially cut sagittal sections of paraffin-embedded sella turcica tissue blocks from 88 normal and pathological fetuses, 13 to 24 weeks of gestation, were examined. Twenty-seven specimens out of 88 had visible notochordal remnants in the cranial base, and these constituted the material available for this study. A straight notochordal course is always seen in normal sella turcica morphology, and a non-straight notochordal course is always seen in malformed sella turcica. Among the fetuses diagnosed at autopsy as "normal fetuses," both normal and pathological findings in the sella turcica regions were observed. The pathological findings were always found in the spontaneously aborted fetuses (five cases). Among the fetuses diagnosed at autopsy as "pathological fetuses," both normal and pathological findings were also observed in the sella region. Our conclusion is that the morphological appearance of the notochordal remnants in the dorsum sellae is associated with the morphology of the sella turcica. These structures ought to be analyzed on larger materials of both normal and pathological fetuses. One of the more obvious perspectives opened up by the present study is the probable disclosure of malformations in spontaneously aborted fetuses without external malformations. PMID- 9338854 TI - Finite element morphometry of the midfacial complex in subjects with Angle's Class III malocclusions. AB - The purpose of this study was to determine whether the morphology of the midface differed in normal (Class I) and midfacially-retrognathic (Class III) prepubertal subjects, and to localize differences morphometrically. Lateral cephalographs of 133 European-American children between 5-11 years of age were traced and average geometries, scaled to an equivalent size, were generated based upon seven nodes (pterygoid point, PTS; rhinion, RO; posterior nasal spine, PNS; midpalatal point, MPP; anterior nasal spine, ANS; subspinale, A; and prosthion, Pr). The samples also were subdivided into seven age- and sex-matched groups for morphometric comparisons. Procrustes analysis indicated that the overall midfacial configurations differed statistically (P < 0.05). Therefore, a color-coded finite element (FEM) program was used to localize differences in morphology graphically. Comparing Class I and III groups for size-change, FEM revealed that negative allometry was evident in the posterior half of the midfacial configuration localized between PTS, PNS, and MPP. The anterior half was more isotropic, however, but the anterior-most aspect of the configuration between Pr and RO showed some positive allometry particularly in the premaxillary and incisor regions. For shape-change, major differences in shape over the entire midface were not as evident, with an isotropic midfacial morphology for normal and Class III subjects. It is concluded that an identifiable pattern of deformation is evident for the Class III subjects during the prepubertal growth period. Therefore, midfacial retrognathia associated with Class III malocclusions results, at least in part, from deficient anteroposterior elongation of the midfacial complex allied with deformation of the premaxillary region. PMID- 9338855 TI - Osteogenesis imperfecta: clinical, cephalometric, and biochemical investigations of OI types I, III, and IV. AB - The aim of the study was to analyze craniofacial development in 54 patients with osteogenesis imperfecta (OI), who were classified into OI types I, III, and IV according to clinical criteria, and to relate the findings to the abnormalities in collagen I production. In 33 patients, analysis of radioactively labelled procollagen was performed. Cephalometric radiographs, facial photographs, and CT scans (a single case) were analyzed and mean facial diagrams for lateral and frontal films were produced based on registration of 221 reference points. Radiographs of 102 male and 51 female Danish students served as control material. In OI type I, size of the skull and jaws was generally slightly reduced, but morphology was within normal limits. In OI type IV and especially type III more severe abnormalities were found; the cranial base was flattened, the maxilla posteriorly inclined, and nearly all size-measurements were reduced. In OI type III the sagittal jaw relations were reduced and a mandibular overjet recorded. Three OI type I patients, whose fibroblasts produced structurally abnormal collagen I, had the stature and several features in the craniofacial region, which corresponded to those recorded for the OI type IV group. Also, in three OI type IV patients whose fibroblasts produced a reduced amount of normal collagen I, craniofacial morphology showed several features resembling type I patients. We conclude that structural abnormalities of collagen I generally give rise to more severe alterations of the craniofacial features than a quantitative defect of collagen I. OI type I patients are only slightly affected in their craniofacial region, while patients with OI type IV and especially type III are moderately to severely affected. The combined cephalometric and biochemical findings suggest that future classification of patients with osteogenesis imperfecta should be based on biochemical/molecular and radiological analyses in combination with clinical criteria rather than on clinical features alone. PMID- 9338856 TI - Characteristic dental arches and occlusion in patients with aspartylglucosaminuria. AB - Aspartylglucosaminuria (AGU) is a lysosomal storage disorder with progressive mental retardation as a presenting manifestation. The disorder is caused by a single nucleotide change in the gene encoding aspartylglucosaminidase (AGA). This rare disease is relatively common in Finland: we were able to examine 81 Finnish AGU-patients for dental and oral changes. Tooth crown size and crown shape were normal, but dental malocclusions were common, and prevalences of spacing, large overjet, anterior open bite, and lateral crossbite exceeded Finnish population prevalences (P < 0.0001). Dental arches were already large in childhood, and in adult patients, when compared to Finnish population standards, the lower dental arch was larger in all dimensions (P < 0.001). Almost all patients had abnormally large tongues, which we assumed to be the reason for the structural abnormalities observed. PMID- 9338857 TI - Association between non-right-handedness and cleft lip with or without cleft palate in a Chinese population. AB - The etiology of non-syndromic cleft lip with or without cleft palate (CL +/- P) is unclear, although both familial and environmental factors are implicated. Because CL +/- P occurs at approximately the time of brain lateralization and is most often unilateral, developmental asymmetry effects have been postulated in CL +/- P etiology. Handedness is frequently used as an indicator of brain lateralization; therefore, several studies have examined the relationship between cleft laterality and handedness. However, these studies have had conflicting results. The present study investigated handedness in a Chinese sample of 211 non syndromic CL +/- P surgical probands (ascertained in Shanghai for family studies of CL +/- P), 221 population-based but unmatched controls, and 272 first-degree relatives of the probands. Handedness was assessed by means of laterality quotients (LQ) calculated from questionnaire data. Mean LQ's were compared, as were various arbitrary definitions of handedness based on the LQ, for cases versus controls, males versus females, right-sided versus left-sided clefts, and cleft lip alone versus cleft lip plus cleft palate. CL +/- P cases had a significantly higher proportion of non-right-handedness (NRH) than controls, regardless of the definition of NRH (P values < or = .001). There were no statistically significant differences for any of the other comparisons. Familiarity of NRH was tested by comparing first-degree relatives of cases to controls; first-degree relatives were found to have a significantly higher proportion of NRH than controls, supporting familial effects in NRH. These results support the concept of a common etiology and/or developmental pathway for CL +/- P and handedness. PMID- 9338858 TI - A longitudinal study of the postnatal maternal effect on the craniofacial growth of mouse offspring by cross-nursing. AB - A cross-nursing experiment was conducted to examine the nursing dam strain effect on the postnatal growth of a newborn mouse offspring by simultaneously using larger DDD strain mice and smaller C57BL strain ones. A periodical cephalometric observation of the postnatal craniofacial growth of the offspring was longitudinally made from birth up to the 100th day of life in addition to measuring the offspring weight. According to multivariate statistical analyses, the following results were obtained: 1) The mean body weight of the DDD offspring cross-nursed by the C57BL dam was significantly lighter than that of the DDD offspring self-nursed by the DDD dam throughout the whole experimental period except at birth and on the 100th day. 2) The mean body weight of the C57BL offspring cross-nursed by the DDD dam was significantly heavier than that of the C57BL offspring self-nursed by the C57BL dam from the 10th day up to the 30th day. 3) There was a significant positive relationship between the offspring weight and the offspring craniofacial size throughout the entire experimental period in the C57BL offspring and from the 10th day up to the 40th day in the DDD offspring. 4) The nursing dam strain effect on the craniofacial size of the DDD offspring was also significant on the 30th, 60th, 80th, and 100th days. The adjusted craniofacial size of the DDD offspring cross-nursed by the C57BL dam was significantly smaller than that of the DDD offspring self-nursed by the DDD dam on the 30th, 60th, 80th, and 100th days. 5) The nursing dam strain effect on the craniofacial size of the C57BL offspring was not significant throughout the whole experimental period. The adjusted craniofacial size of the C57BL offspring cross nursed by the DDD dam was almost the same as that of the C57BL offspring self nursed by the C57BL dam throughout the whole experimental period. 6) The fat content of the DDD dam tended to be higher than that of the C57BL dam and the interstrain difference was closed to a significant level (P = 0.06) on the 20th day. The moisture content of the DDD dam tended to be lower than that of the C57BL dam and the interstrain difference was also significant on the 7th day (P < 0.001). Based on these findings, it can thus be concluded that the nursing dam strain effect played an important role in the postnatal somatic growth of the DDD and the C57BL offspring and the craniofacial growth and the DDD offspring. PMID- 9338859 TI - Intimate justice: confronting issues of accountability, respect, and freedom in treatment for abuse and violence. AB - Intimate justice theory is a set of nine interrelated concepts that describe the ethical dimensions of equality, fairness, and care in ongoing partnerships. Understanding ethical dimensions involves examining internalized beliefs and behavior in terms of their motivation and impact on the partner, particularly as they empower, disempower, or abuse power. The concepts of intimate justice theory are applied to confront disempowerment and abuses of power, to challenge internalized beliefs about how one should treat one's partner, to explore how internalized beliefs were developed through experiences in the family of origin, and to develop an awareness of the linkages between intimate partner abuse and social injustice. This article demonstrates how therapists can utilize three of the concepts --accountability, respect, and freedom--to structure the opening phase of treatment for abuse and violence. The primary focus of the opening phase is on establishing accountability for change in the abusive man and protecting the safety of the injured partner. This involves challenging the abuser's sense of entitlement and working to rethink what respect is and restoring freedom to his partner. The discussion incorporates the findings of an exploratory, qualitative study that investigated the experiences of 30 abusive men and their partners who were clients in a university-based counseling clinic. The article elaborates six interventions that can be utilized in clinical settings to structure treatment with abusive men. PMID- 9338860 TI - The politics of gender in family therapy. AB - This article provides an overview of the political implications of various approaches to gender within the clinical literature. It emphasizes the process of therapy within the social context of gender relations and identifies the political consequences of various clinical responses. Issues surrounding the appropriate role and stance of therapists relative to gender are identified, ethical issues such as neutrality and client welfare are re-examined, and suggestions for practice are addressed. PMID- 9338861 TI - Curriculum changes to meet challenges: preparing MFT students for managed care settings. AB - Fledgling therapists who graduate from family therapy training programs will have to navigate the world of managed care. In this article, faculty of University of San Diego share changes in its accredited training program that prepare students for practice in an increasingly multidisciplinary world where health maintenance organizations and other versions of managed care predominate. The paper touches on contextual issues, provides a detailed outline of coursework presenting basic knowledge and skills involved in clinical practice in a managed care environment, and comments on clinical placements and the challenge of helping the next generation of clinicians "fit" into the future of health care delivery while maintaining their unique identity as family therapists. PMID- 9338862 TI - Beyond law and ethics: an interdisciplinary course in family law and family therapy. AB - The professions of family therapy and law share many clients and areas of overlap. Law-related coursework in family therapy programs is typically limited to legal, ethical, and professional issues. However, students can also benefit from understanding other areas of overlap, such as divorce, child custody, and mediation. This article discusses the curriculum for an interdisciplinary course that educates both family therapy and law students. The course provides: (1) a substantive education about similarities and differences between the professions, how they operate as systems, and specific areas of overlap, (2) opportunities to learn clinical skills, and (3) opportunities for personal insight about skills, personality types, and negotiation styles, and how these may differ between the professions. PMID- 9338863 TI - Dynamic biologic transformation of the periodontium: a clinical report. AB - The technology of dynamic biologic transformation of the periodontium differs from orthodontic therapy in that the primary goal of dynamic biologic transformation of the periodontium is to reshape an abnormal periodontium. The goal of orthodontic treatment usually is the straightening of teeth in the orofacial complex. The new term dynamic biologic transformation of the periodontium has been coined to differentiate between a change in the periodontium that is secondary to straightening teeth through usual orthodontic methods to one that the primary goal is the rearrangement of the periodontium in which the tooth is merely used as a handle to pull the periodontium into the desired shape and location. The tooth or teeth that are moved in this process are often reshaped or put into abnormal positions to accomplish this goal. A review of the literature revealed that these changes in the periodontium can be used in various manners that will lessen surgical morbidity and maximize esthetics in conjunction with restorative and periodontal care. Combinations of dynamic biologic transformation of the periodontium and surgery were shown through clinical reports to illustrate this principle. PMID- 9338864 TI - Minimizing prosthesis movement in a midfacial defect: a clinical report. PMID- 9338865 TI - Longitudinal study of pressed glass-ceramic inlays for four and a half years. AB - STATEMENT OF PROBLEM: Some restorative materials used in the posterior region of the mouth present conventional problems, such as microleakage, recurrent caries, wear, and poor color stability. PURPOSE: This study determined the reliability of the IPS Empress ceramic material for fabricating inlays and onlays in the posterior region of the mouth. MATERIAL AND METHODS: A total of 125 IPS Empress pressed glass ceramic inlays were placed for 29 patients in a private practice. The restorations were observed for a period of 7 to 56 months, with a mean of 40.3 months. All inlays were constructed in the same dental laboratory and the restorative materials used according to the manufacturers' instructions. The restorations were evaluated clinically according to modified U.S. Public Health Service criteria at the time of insertion and at periodic recall appointments. Kaplan-Meier statistics were used to calculate survival rates. RESULTS: Clinical evaluation revealed that pressed glass ceramic inlays, with the exception of four fractured restorations, were rated from Alpha to Bravo for each criteria. Marginal discoloration recorded the lowest percentage of alpha ratings (65.3%). The estimated survival rate after approximately a 4.5-year follow-up period was 95.63% (95% confidence interval; 90.77% to 99.95%). PMID- 9338866 TI - Adherence of "stuck" restorations to demineralized dentin following use of experimental primers. AB - PURPOSE: This in vivo study histopathologically evaluated the biocompatibility of four self-conditioning dentinal primer formulas in four bonding systems and evaluated the pulpal responses of the self-conditioning dentinal bonding systems for Class V tooth preparations in primates. MATERIAL AND METHODS: The basic formula consisted of proprietary carboxylic diacid monomer, dipentaerythritol pentaacrylate phosphoric acid ester, acetone, and ethanol that was used alone or mixed with other commercial priming agents. After application of primers, Prisma Universal Bond 3 was selected to restore the cavity preparations. RESULTS: All systems were judged histopathologically biocompatible. "Stuck" restorations that were resistant to acid demineralization necessary for processing of histologic slides, occurred more frequently with intact smear layers and increasing use of dehydrating agents. CONCLUSIONS: Tests should be performed to determine whether "stuck" restorations in in vivo studies that maintain dentin-pulpal relationships can support in vitro shear bond strength tests. PMID- 9338867 TI - The prevalence, etiology and management of tooth wear in the United Kingdom. AB - STATEMENT OF PROBLEM: Recent epidemiologic evidence suggests that tooth wear is now a significant problem in both children and adults. There is growing evidence that a major cause of severe wear in patients is regurgitation erosion due to a variety of factors including gastroesophageal reflux disease. PURPOSE: The purpose of this article is to discuss the prevalence of tooth wear in the United Kingdom. Emphasis in management should be on accurate diagnosis, and in some patients, long-term monitoring before embarking on any irreversible, interventive treatment. Even when treatment is necessary, a period of monitoring is helpful to assess the rate of progress of the wear, the effectiveness of preventive measures, and therefore the extent of the treatment necessary. PMID- 9338868 TI - Reproduction of excursive tooth contact in an articulator with computerized axiography data. AB - PURPOSE: This study assessed the reproduction of excursive tooth contacts with a SAM2 "P" articulator set up with the aid of computerized axiography. MATERIAL AND METHODS: Articulator excursive tooth contacts were compared with intraoral excursive contacts identified with occlusogram wax. The maxillary casts were mounted with a kinematic face-bow, and the mandibular cast was oriented to the intercuspal position. The articulator was set using information obtained from computerized axiography. Protrusion and laterotrusion to the left and right sides were examined. The first 4 mm of excursive tooth contacts in the occlusogram were compared with the 4 mm of excursive tooth contacts in the articulator. RESULTS: The articulator reproduced 82% of the teeth with protrusive tooth contacts and 90% of the teeth with laterotrusive tooth contacts. The exact locations of excursive tooth contacts were reproduced in 66% of protrusive contacts and 81% of laterotrusive contacts. The articulator also created additional eccentric dental contacts that were not originally present in the occlusogram. Twenty percent of the subjects showed these additional contacts during protrusion; in lateral movement of the articulator these additional contacts were present in 27% to the left side and 20% to the right. Clinically, these findings suggest that there are limits to the ability of the articulator to reproduce excursive tooth contacts. These limitations should be kept in mind when an articulator is used for diagnostic and restorative dental procedures. PMID- 9338869 TI - Preliminary results of a multicenter study evaluating a chemically enhanced surface for machined commercially pure titanium implants. AB - PURPOSE: This report presents the preliminary results of a prospective multicenter study to evaluate the efficacy of a chemically etched pure titanium surface on screw-type dental implants. MATERIAL AND METHODS: The results of 147 implants placed in 75 patients were evaluated and followed for up to 3 years, using clinical and radiographic examinations. The implants were inserted to support single crowns, overdentures, and fixed prostheses, as dictated by the individual patient's need. RESULTS: Of the 147 implants placed, 5 were lost and the remaining implants were followed throughout, to the end of the study period. Crestal bone levels adjacent to each implant were monitored and the amount of total bone loss was calculated. Clinically healthy peri-implant gingival tissues were observed on 95% of the implants, whereas 88.3% demonstrated no bleeding on probing and no recession during the follow-up period. According to the criteria used in this study, the total success rate was calculated to be 96.6%. PMID- 9338870 TI - Professional hygiene care, adjustments and complications of mandibular implant retained overdentures: a three-year retrospective study. AB - PURPOSE: This report presents a retrospective evaluation of postinsertion care required by 104 edentulous patients with advanced mandibular bone loss. MATERIAL AND METHODS: The patients were treated with new maxillary dentures and mandibular overdentures retained by two implants with a single bar-clip attachment. Distinction was made between professional hygiene care, adjustments, and treatment of complications. The follow-up period after insertion of dentures was 3 years for all patients. RESULTS: Approximately a third of the patients needed professional hygiene care. The need for adjustments declined during the years of function. Complications were encountered in approximately a third of the patients. The majority of these were not related to the implants, but to the superstructure and both the maxillary and mandibular dentures. CONCLUSIONS: Many edentulous patients with advanced mandibular bone loss who were treated with mandibular implant-retained overdentures need professional hygiene care, adjustments, and treatment of complications. PMID- 9338871 TI - Theoretical study of the effects of tooth and implant mobility differences on occlusal force transmission in tooth/implant-supported partial prostheses. AB - STATEMENT OF PROBLEM: Despite their mobility differences under occlusal loads, a natural tooth and an implant are often used together to support fixed prostheses. In some situations, tooth/implant-supported partial prostheses include cantilever extensions, especially in the posterior region where the bone is inadequate for placement of an additional implant. PURPOSE: In this study, engineering beam theory was used to study the effects of the mobility differences between the implant and the tooth on the force and moment distribution, due to occlusal loads in tooth/implant-supported prostheses. METHODS: The prosthesis was treated as a linear elastic beam and the supports were modeled as springs with (vertical) translational and rotational stiffness. The bending moments and forces on the supports were calculated as functions of the parameters that describe the geometry, position of the occlusal load, and stiffness ratios (namely, implant or tooth) of the springs. RESULTS: Bending moments on the supports were more sensitive to the relative rotational mobility between the supports and their individual values than to the relative translational mobility. The moment at the implant was minimized when the supports had similar mobilities. A preliminary design concept was introduced and eliminated the moment at the implant without significantly increasing the magnitude of the moment at the tooth. Cantilevering the prosthesis resulted in moderately increased bending moments and considerable tensile forces on the supports for a broad range of the parameters that describe the geometry and loading. CONCLUSIONS: From this simulation, it is suggested that cantilever extensions should be avoided or supported by a short implant, which will only restrain the vertical movement of the cantilever end. PMID- 9338872 TI - The effects of sprue design on the roughness and porosity of titanium castings. AB - PURPOSE: This study measured the effects of the sprue number and position on the roughness and porosity of cast titanium crowns. MATERIAL AND METHODS: Twenty-four complete veneer crown wax patterns were fabricated on a stainless steel die with a 30-degree bevel finish line. Twelve wax patterns were sprued with one 8-gauge wax sprue and the remaining 12 were double sprued. All patterns were invested with a phosphate bonded investment. Castings were made with a titanium casting, according to the manufacturer's instructions, with commercially pure titanium (> 99.5%) ingots. The castings were carefully cleaned and the surface roughness was measured with a profilometer. The specimens were then embedded and sectioned. Internal porosities were quantified with photographs by computerized image analysis. Data were analyzed with an ANOVA and the Student's t test with a confidence level of 95%. RESULTS: The roughness value of the occlusal third of the crowns for the single sprue group (Ra = 3.0 +/- 0.9 microns) was significantly higher than other measurements (p < 0.05). There were statistically significant differences in values of porosity areas between the single sprue group (1.5 +/- 0.7 mm2) and the double sprue group (0.2 +/- 0.2 mm2) (p < 0.01). The double sprue design resulted in a relatively smoother casting surface and less internal porosity than the single sprue design. CONCLUSIONS: Improvements in the degree of roughness and porosity of titanium crown castings were the result of the double sprue design. PMID- 9338873 TI - A simplified procedure for stabilizing pontics. AB - A simplified technique is described for use when waxing pontics to a fixed partial denture which ensures accurate pontic placement without attaching the pontic to the retainers until late in the fabrication process. This technique allows easy access to the proximal contours and margins of the retainers and to proximal and gingival contours of the pontic while maintaining an accurate occlusal relationship. PMID- 9338874 TI - Resin-bonded metal-ceramic inlays: a new approach. AB - With increasing demand for esthetics, dentists face the challenge of delivering definitive restorations that fulfill patients' expectations of esthetics, biocompatibility, and durability. Recent technical developments have encouraged fabrication of gold-reinforced porcelain inlays that meet these important criteria. This article describes a sequence to construct metal-reinforced porcelain inlay restorations. PMID- 9338875 TI - Surveying removable partial dentures: the importance of guiding planes and path of insertion for stability. AB - STATEMENT OF PROBLEM: Although removable partial dentures are a favored option for the restoration of many situations that involve partial tooth loss, some patients are not satisfied with a removable partial denture, especially when it is not stable during mastication. A dental surveyor can be used to prevent countless problems related to the production of removable partial dentures. Many professionals working with oral rehabilitation fail to take advantage of the many uses of a surveyor in planning and designing chromium alloy and other metal removable partial denture frameworks. PURPOSE: This article uses an academic approach to describe the criteria used to determine the path and removal of a removable partial denture. A fundamental requirement for understanding the correct use of the dental surveyor is to prevent indiscriminate use of a path of insertion perpendicular to the occlusal plane, and extreme inclinations of the cast in the attempt to create undercuts on some teeth. PMID- 9338876 TI - The distortion of cast cobalt-chromium alloy partial denture frameworks fitted to a working cast. AB - PURPOSE: This in vitro study examined the distortion of 10 identically designed cast cobalt-chromium alloy frameworks, individually constructed, and adjusted by 10 technicians to fit one of 10 replicated stone working casts. MATERIAL AND METHODS: The distortion was measured with a specially developed strain gauge apparatus when the frameworks were seated on the maxillary metal analog, the stone working cast, or a stone duplicate cast. RESULTS: A statistically significant difference between the distortion of each of the frameworks was observed. A significant difference in the distortion was also observed for the frameworks when placed on each of the three casts. The distortion was the least when the frameworks were seated on the stone working cast and the greatest when seated on the metal analog. CONCLUSION: This study demonstrated that the distortion of a framework is not related to its closeness of fit as observed by the presence or absence of gaps between the framework components and the teeth. PMID- 9338877 TI - Anterior margin adaptation for implant-retained auricular prostheses. AB - Adaptation of the anterior margin of implant-retained auricular prostheses may be compromised during mandibular movement. Loss of contact between prostheses and underlying skin in an esthetic area is of great concern. This article describes an impression procedure that addresses the problem of anterior margin adaptation. PMID- 9338878 TI - A substitute for facial replica in mannequin procedures. AB - The use of mannequin exercises has been a teaching tool in the education of dental students. Various commercially available materials to simulate extraoral facial structure form part of this apparatus, also called a "phantom head." It is not always practical, with cost considerations, to replace this external component when unavoidable wear and tear occurs. An alternative that uses a recycled milk carton is presented as an inexpensive substitute. PMID- 9338879 TI - Securing the abutment post screw in a single implant prosthesis. PMID- 9338880 TI - A method of examining the magnitude and origin of "soft" and "hard" tissue forces resisting limb lengthening. AB - Complications arising from limb-lengthening procedures such as muscle contracture, axial malalignment of the bone and traction injuries to the nerves and vessels, are often severe. Often complications arise from the build-up of forces in the biological tissues which are resisting lengthening. Little is known about the origin and magnitude of these forces, although three studies have identified the regenerate (new bone tissue) as the dominant resisting tissue. This study describes the development of a method to examine these forces. It employs load measurement devices in the structural columns of Ilizarov fixators which measure the compressive load on the frame exerted by the biological tissues. The distribution of this load between the columns of the frame, in conjunction with a transverse radiograph of the limb at the regenerate site, is used to examine the origin of the resisting force. Accuracy was determined by a laboratory simulation which found the predicted position of the force to be within 5 mm of the actual position in all four cases tested. Mean error in the total measured force was 2 N (SD, 1 N). A pilot study on a patient undergoing a 60 mm femoral lengthening revealed a peak force of 717 N originating in the Vastus Lateralis or the illiotibial tract. Negligible contribution to resistance was provided by the regenerate, contrary to that found with other studies. PMID- 9338881 TI - Methodology and apparatus to determine material properties of the knee joint meniscus. AB - This paper describes a soft tissue test method and apparatus which determines the constants necessary to characterise the elastic properties of a transversely isotropic material such as the knee joint meniscus. The tensile machine was designed to test small delicate samples in a humidified environment and measure specimen deformation with a CCD video camera. The method described here employs a common specimen preparation procedure and test protocol using specimens with four different fibre orientations. Discrete markers (0.075 mm graphite particles) permit repeatable measurements of two-dimensional strain on the specimen surface enabling the determination of Poisson's ratio characteristics. Shear strain was also measured in tensile test specimens prepared with an oblique collagen fibre orientation, offering an alternative method to investigate shear properties. Regional strain measurements provide an assessment of the uniformity of the tissue deformation. Preliminary results on bovine menisci are presented as an application of the test system. Material properties characterising the meniscus tissue were obtained with one test procedure and identical coupon size. This has the advantage of avoiding systematic errors caused by different preparation techniques, test procedures and equipment. PMID- 9338883 TI - Mechanical simulation of composite hip stems. AB - Calculation of new orthopedic implants prior to manufacturing of prototypes can be economic in the case of complex production processes. The use of composite materials for highly loaded hip stems is one application of the Finite-Element Method. Due to the anisotropic behaviour of composite structures, special routines had to be programmed for element alignment and failure analysis. Stability of carbon fibre-reinforced epoxy hip stems could be confirmed by experimental results. The risk of neck fracture was found to be one of the critical features in the design process. PMID- 9338882 TI - Spatio-temporal representation of multichannel EMG firing patterns and its clinical applications. AB - Analyzing motor unit (MU) activity is essential for studying the neurological dysfunction of upper motor neuron disorders (UMND). This study employs multichannel surface electromyographic (EMG) signals, as recorded from the upper arm during elbow flexion and extension, to analyze the temporal changes and spatial distribution of the dominant firing rate. To estimate the dominant firing rate, the autoregressive (AR) spectrum analysis method is utilized to detect the peaks and poles of the AR model, of the surface EMG spectrum below 40 Hz. The temporal changes in firing rates are also observed by using the spectrogram representation of low-frequency EMG spectra. The EMG spectrogram facilitates examination of the time-varying characteristics of firing rates and recruitment of MUs from surface EMG signal. The low-frequency spectra of multichannel EMG are then represented in a polar form to visualize the spatial distribution of firing patterns across muscles. Via spatio-temporal representation techniques, this study provides a viable approach of observing both the spatial and temporal patterns of MU activities in normal subjects and patients with UMND, including cerebrovascular disease and Parkinson's disease. PMID- 9338884 TI - Ambulatory monitoring of children's activity. AB - One of the difficulties in clinical assessment is how to obtain accurate data in the "real world". This paper describes the Dynaport ADL Monitor, an accelerometry based system for ambulatory monitoring of the activities of daily living (ADL). In previous studies the monitor has proved a success with adults. To validate the system for use with children, 9 h of various activities conducted by nine children were measured and videotaped at the same time. All postures and activities were divided into one of five main categories: standing, sitting, lying, locomotion and swing/seesaw. The video pictures were evaluated by an observer. Independent of this, the acceleration signals were translated by the DynaPort ADL monitor software, and were compared to the video pictures. Minimal and maximal validity percentages were calculated for each of the main classes, for each individual measurement and overall. To estimate monitoring performance the 10 sets of measurements are regarded as representative samples of children's daily activity. The overall minimal and maximal validity are 73.15 +/- 1.96 x 4.48 and 91.31 +/- 1.96 x 1.75 weighed standard deviation. PMID- 9338885 TI - An experimental study of the failure modes of the Gamma Locking Nail and AO Dynamic Hip Screw under static loading: a cadaveric study. AB - The sliding compression screw is widely regarded as the optimum treatment for intertrochanteric fractures of the femur, allowing bone fragments to impact until a bony support has been established across the fracture site. This study carried out biomechanical, cadaveric tests to establish the influence of direct static loading situations on the modes of failure of the Gamma Nail compared with the Dynamic Hip Screw (DHS). Clinical studies report DHS failures of lag screws cutting-out, bending of the lag screws and cortical screws pulling out causing plate loosening. Gamma Nail failures include lag screw cut-out or fractures of the femoral shaft around the distal locking screws or nail tip. In this study each failure mode has been isolated, to establish the loads to failure under various fracture configurations. The biomechanical results indicated that the intramedullary Gamma Locking Nail can be recommended over a standard DHS in cases of subtrochanteric fracture or conditions of very poor bone quality. PMID- 9338886 TI - Computer prediction of adaptive bone remodelling around noncemented femoral prostheses: the relationship between damage-based and strain-based algorithms. AB - Several mathematical models to predict tissue adaptation have been derived since Julius Wolff proposed a function-form relationship for bone. These can be formulated as computational procedures (algorithms) to predict bone adaptation around implants. The objective of this paper was to further develop the damage adaptive algorithm, to test its validity, and to determine the relationship between it and algorithms based on strain energy. This was achieved using finite element models of the proximal femur, one for the intact case and another for the case where a noncemented hip prosthesis is implanted. The finite models were generated using CT scan data. Initial bone resorption patterns around a femoral prosthesis following total hip arthroplasty were computed for both damage adaptive and strain-adaptive adaptation rules. It is found that the damage adaptive algorithm can successfully predict the bone's adaptive behaviour in response to altered mechanical loading provided that account is taken of the nonlinear nature of damage accumulation. Predictions are made using a strain energy stimulus for comparison with the damage stimulus, and a theoretical relationship between the two is proposed. It is shown that an advantage of the damage approach over the strain-based approach is that the nonlinearity required to replicate clinically observed resorption patterns can be derived theoretically, whereas for strain-adaptive remodelling, empirical relationships are assumed. PMID- 9338887 TI - The effects of density and test conditions on measured compression and shear strength of cancellous bone from the lumbar vertebrae of ewes. AB - An animal model (the ewe) was used to study mechanical parameters of cancellous bone specimens. Compression and shear tests were conducted on ewe vertebral trabecular bone (L1-L5) from old ewes (mean age: 9 years) under two different conditions: first, at room temperature in air ("standard" test conditions); and secondly, in a physiological saline bath regulated at 37 degrees C. The parameters obtained under "standard" test conditions with a uniaxial compression test were the mean value of the maximum strength (sigma max = 22.3 (7.06) MPa), Young's modulus (E = 1510 (784) MPa), the strain at maximum strength (epsilon sigma max = 3.21 (0.8) percent) and the energy absorbed during the test (W = 0.3 (0.12) MJ.m-3). No significant change was found when the test was carried out in a saline bath at 37 degrees C (p < 0.0005). An original shear test was performed to evaluate the shear strength which was found to vary from 7.5 (4.7) to 14.6 (8.53) MPa under "standard" test conditions depending on the method of calculation. Testing of the specimens in a 37 degrees C physiological saline bath induced a decrease in the shear strength from 32.5 percent (p < 0.0005) to 37.3 percent (p < 0.0001) of those measured under "standard" test conditions. The non destructive measurement of the Bone Mineral Density (BMD) accounted for up to 73.3 percent of the maximum compressive strength sigma max and 61.5 percent of the maximum shear strength tau max determined in saline solution at 37 degrees C. These results showed that other parameters influencing the mechanical properties of trabecular bone and its structure appeared to be essential. PMID- 9338888 TI - Determination of transients and compensation capacities of breath-by-breath analysis by cubic splines. AB - The development of breath-by-breath analysis during an ergospirometry improved the precision of the measurement. However, the abundance of data yields oscillating curves which make it very difficult to detect exactly the breakpoints, maxima and minima. By using cubic splines one is able to smooth the curve of the primary data without falsifying or distorting it. A breakpoint marks the beginning of a hyperventilation with an nonlinear increase of VE or the beginning of an excess value of CO2. Furthermore, the amount of CO2 required to compensate for the acid-base balance as well as the oxygen debt in the recovery phase can be calculated by the area under the curve. PMID- 9338889 TI - Nonlinear high pass filter for R-wave detection in ECG signal. AB - A simple and easily implemented method for R-wave detection from ECG signals is presented. The method is based on the subtraction of a filtered version of the signal. The filter we used is a nonlinear median filter. The median filter is applied to the ECG signal. It results in a smoothed version of the signal, without any reminder of the R waves. This smoothed signal is then subtracted from the original and the resulting signal presents undistorted R-waves, without baseline drift. A simple threshold detection can then be performed on the filtered signal. Results are presented for simulated signals, with sinusoidal and step baseline drifts, as well as ECG complex shape change. The detection is accurate and the average error we obtained for 300 s length signals was of the order of 10(-8)s, for RR intervals of 1 s. Results are also presented for a real experimental signal with strong baseline drift, and the accuracy of the detection can be observed. PMID- 9338890 TI - Power spectra accuracy improvement by a method for leakage effect reduction. AB - A method for leakage effect reduction by a modification of the classical spectra obtained Discrete Fourier Transform is proposed. The properties of the new method are investigated analytically and validated through synthesized signals, processed by the developed complete computational procedure. This combines the proposed method for leakage reduction and our previous approach for optimal epoch selection, enabling more accurate spectral analysis of relatively short epochs of various biomedical and other signals. An application of heart rate variability to power spectra is presented. PMID- 9338891 TI - Application of the design of experiments for the evaluation of the robustness of video-densitometric measurements. PMID- 9338892 TI - Classics revisited: the ego in anxiety (Max Schur, 1953) and, an addendum to Freud's theory of anxiety (Charles Brenner, 1953). PMID- 9338893 TI - Masochism and the inner mother. AB - My understanding and treatment of patients with severe masochistic features has evolved and changed radically over the last fifteen years. My work with John and his subsequent life led me to search for new techniques and eventually to the discovery of a different way of understanding and treating these patients. I analyzed John in accord with the classical Freudian formulation of masochism. The paternal transference was central and-while a maternal transference and work with the residues of his experience with his mother were present and partially analyzed-the real power of the inner mother was not touched. It was only when I saw him years later in a near-moribund state that I realized the centrality of what we had not analyzed: the attachment to a punishing inner mother. This led me to conceptualize masochism in a new way. Of central importance in the formation of severe masochism is the relationship between an indifferent, possessive, or rejecting mother and a helpless child in the earliest years, before object constancy. This is a time when the child is unable to differentiate between self and mother. What results is a preverbal conviction that they are ungrateful or "bad" if they think, feel, or behave differently than the mother. This leads to a powerful and rigid attachment to this early mother, internalized as a punishing inner mother. This is the precursor of masochism, not a regression from the Oedipus complex. The child will tolerate physical and mental suffering to remain attached to the needed-even though pain inducing-mother. If the child is not attached, he feels helpless and fears survival. This attachment and fear is internalized and becomes unconscious as development proceeds. Eventually, what is observed in adult patients is a person who is sensitive to others, but unable to be sensitive to him/herself. Awareness of the importance and power of the attachment to the punishing inner mother enables the analyst to hear and perceive masochistic material with a shifted focus. This shifted focus naturally leads to different psychoanalytic treatment techniques throughout the analytic process. Two other cases of masochism are used to illustrate the techniques and course of their analytic processes with this "inner mother" focus. The psychoanalytic understanding of many forms of disturbance has shifted from the oedipal period to the early years in recent years. There is also a corresponding emphasis on the actual experiences with early figures, most prominently the mother. The particular dynamics I have described is part of this general trend. PMID- 9338894 TI - The minyan as a psychological support system. AB - Most individuals participate in some of the rituals and/or regular activities of religious institutions such as churches or synagogues. Through such involvements, people are offered vital assistance in dealing with developmental changes, opportunities for personal development and for group support, and more generally, a sense of continuity and of meaning in life. This paper deals with only one small aspect of Jewish observance, an aspect of the centuries-old required weekly prayer groups-the minyan. The prime emphasis resides in the rarely recognized, nonliturgical dimension of this small group experience. Using psychoanalysis in the sense of a general psychology as background, I have considered the minyan as combining elements of a psychological support system and of a small group. In addition to the gratification of affiliative needs (social hunger) and the countering of loneliness and of isolation, this group experience helps its members maintain an intergenerational sense of personal identity and of self esteem. In the face of marked life stressors such as death in the family, religious institutions such as the minyan, with its prescribed ritual steps for grieving (i.e., kaddish), fulfill especially significant preventive and restitutive mental health functions. I have also hypothesized that on a covert, fantasy level, the caring and nurturing family-like weekday minyan may even represent a mother-symbol (mother group) in line with people's universal need to establish a psychological union with others, thus restoring an earlier, conflict free state of the child-mother bond. In an extended societal sense, the earlier emphasis in Western cultures on the virtual worship of individuality, autonomy, and independence has given way recently to a renewed appreciation of cooperation, communalism, and altruism. The minyan, as a small religious communal aggregate with its inherent climate of mutuality, reciprocity, and continuity, has, in a sense, anticipated these new developments. For as Herbert Simon (1990), the Nobel laureate, observed, "It is of no little moment for the human future whether people are necessarily and consistently selfish...or whether there is a significant place for altruism in the scheme of human behavior." PMID- 9338895 TI - "History is not the past": Lacan's critique of Ferenczi. PMID- 9338896 TI - Figures of entrapment: the labyrinth and the web. PMID- 9338897 TI - The Gambler as case history and literary twin: Dostoevsky's false beauty and the poetics of perversity. PMID- 9338898 TI - Food as selfobject in eating disorder patients. PMID- 9338899 TI - Back pain: beguiling physiology (and politics) PMID- 9338900 TI - Does combined spinal-epidural analgesia with subarachnoid sufentanil increase the incidence of emergency cesarean delivery? AB - BACKGROUND AND OBJECTIVES: The purpose of this review was to determine if patients who receive combined spinal-epidural (CSE) analgesia with subarachnoid sufentanil have an increased incidence of emergency cesarean delivery for fetal distress when compared with patients who receive systemic or no medication (S/NM) for labor analgesia. METHODS: A retrospective computerized analysis of data on all 2,560 deliveries at Bellevue Woman's Hospital for 14 months summarized practice parameters for 1,240 patients who received regional analgesia (98% CSE analgesia), identified 1,140 patients who received S/NM, and classified the urgency of 479 cesarean deliveries. In the CSE group there were 168 cesarean deliveries (emergency 16, urgent 58, semiurgent 70, and nonurgent 24) as compared with a total of 128 (emergency 16, urgent 43, semiurgent 69, nonurgent 0) in the S/NM group. Scheduled cesarean sections (180) were excluded from the study. RESULTS: The incidence of emergency cesarean delivery in 1,217 patients who received CSE analgesia with subarachnoid sufentanil (10-15 micrograms) compared with 1,140 patients who received S/NM for labor analgesia was 1.3% versus 1.4%, respectively. More importantly, there was no case in which emergency cesarean delivery was required for acute fetal distress in the absence of obstetric factors during the 90 minutes following administration of subarachnoid sufentanil. General anesthesia was required for emergency cesarean delivery in only one patient (6%) in the CSE group, as against eight patients (50%) in the S/NM group who required general anesthesia for emergency cesarean section (P < .05). CONCLUSIONS: This experience indicates that patients who receive CSE analgesia do not have a higher incidence of emergency cesarean delivery than patients who have S/NM for labor analgesia. Emergency cesarean section for fetal distress within 90 minutes of the administration of intrathecal sufentanil only occurred in association with obstetric factors. However, caution should be exercised in extrapolating these results to other practice settings, particularly high-risk referral centers. PMID- 9338901 TI - Combined spinal-epidural technique. AB - BACKGROUND: For the past 16 years the combined spinal-epidural (CSE) technique has been extensively researched and developed to the point where it is now in widespread use. Along with the use of low-dose mixtures of local anesthetics and opioids, and the introduction of fine-gauge pencil-point needles, CSE is being increasingly recognized as another important addition to the armamentarium of the anesthesiologist. METHOD: One hundred and forty-two publications focusing on the CSE technique, or on questions concerning CSE-related issues, were reviewed. Out of 33 double-blind or controlled studies, 23 directly investigated the CSE technique. Fifty-four prospective studies and letters made directly relevant points on advantages or problems with CSE. Twenty-one book chapters, reviews and editorials were included, 11 of them concerning the use of CSE technique. RESULTS: All CSE techniques which may be used for surgical anesthesia, for analgesia in labor or for postoperative pain management are described. Indications, advantages, disadvantages, and the various methods for performing CSE procedures, including sequential CSE block, are described and reviewed, along with the equipment currently available for their administration. CONCLUSION: The CSE technique offers many potential advantages over continuous epidural or subarachnoid methods alone, including a reduction in drug dosage, the ability to eliminate motor blockade and to achieve highly selective sensory blockade and optimize analgesia. These features hold great promise for minimizing the hazards and side effects of traditional epidural and subarachnoid techniques. Controversial fears, risks, and pitfalls of the CSE technique and of continuous epidural and subarachnoid methods are debated and discussed. PMID- 9338902 TI - Does metoclopramide supplement postoperative analgesia using patient-controlled analgesia with morphine in patients undergoing elective cesarean delivery? AB - BACKGROUND AND OBJECTIVES: Recent studies have shown that metoclopramide may decrease postoperative narcotic requirements in patients undergoing second trimester induced abortions or prosthetic hip surgery. It is often used to decrease the incidence of nausea and vomiting in the patient undergoing cesarean delivery under regional anesthesia. If metoclopramide were found to be an analgesic adjunct in these patients, it would offer an additional impetus for its routine use. METHODS: After elective cesarean delivery under spinal anesthesia, 32 patients were monitored for initial and 24-hour postoperative morphine requirements via intravenous patient-controlled analgesia. These patients were divided into two groups. Prior to spinal block, group 1 (n = 17) received 10 mg intravenous metoclopramide, and group 2 (n = 15) received an intravenous saline placebo. RESULTS: No differences were found between groups in the time from spinal placement to the time of pain onset, the amount of morphine necessary to initially achieve comfort, or 24-hour postoperative morphine requirements. (P > .05). CONCLUSIONS: This study demonstrates that metoclopramide decreases intraoperative nausea but does not supplement analgesia in patients undergoing elective cesarean delivery. PMID- 9338903 TI - Culture of bacteria from lumbar and caudal epidural catheters used for postoperative analgesia in children. AB - BACKGROUND AND OBJECTIVES: Continuous epidural analgesia has been used with increasing frequency to provide postoperative pain relief for children. Epidural space infection is a potential complication of epidural catheter placement. This study investigated the incidence of bacterial colonization on lumbar and caudal epidural catheter tips in postoperative pediatric patients. METHODS: In this prospective study, lumbar and caudal epidural catheters were placed in the operating room with aseptic technique. Dilute local anesthetic and/or opioid infusions were used for postoperative analgesia. On discontinuation of the epidural infusion, the skin site was decontaminated with 70% alcohol and then cultured. The distal catheter tip and hub were cultured. Semiquantitative and qualitative aerobic cultures were performed. RESULTS: Data from 91 epidural catheters were available (45 caudal versus 46 lumbar). Of the 45 caudal catheter tips 9 (20%) were colonized, compared with 2 of the 46 (4%) lumbar catheter tips (P < .02). Staphylococcus epidermidis was the predominant skin and catheter tip organism isolated in both groups. Four of nine caudal catheter tips grew gram negative bacteria. Statistical analyses did not show that time, skin site inflammation, or dressing condition were independent predictors of catheter tip colonization. No patient developed a clinical epidural infection during the study period. CONCLUSIONS: The results of this study suggest that the risk of clinical epidural infection associated with caudal or lumbar postoperative catheters is low. However, the incidence of epidural catheter tip colonization is increased with the caudal route of insertion, and the bacteria differ from those cultured from the lumbar insertion site. PMID- 9338904 TI - Adrenocorticotropic hormone infusion as a novel treatment for postdural puncture headache. AB - BACKGROUND AND OBJECTIVES: In two patients, one scheduled for epidural anesthesia and the other for placement of a spinal catheter for operative procedures, severe postdural puncture headache developed and was refractory to conservative therapy. METHODS: The first patient had several unintentional dural punctures, and the second underwent a planned dural puncture with an 18-gauge needle for insertion of a 20-gauge catheter. When neither patient responded to conservative therapy following development of postdural puncture headache, an infusion of adrenocorticotropic hormone (ACTH) was given prior to consideration of epidural blood patching. RESULTS: Both patients obtained complete and permanent relief from their headaches. CONCLUSION: A single treatment with ACTH may offer an alternative therapy in the treatment of postdural puncture headache. PMID- 9338905 TI - Combined general and epidural anesthesia versus general anesthesia for major abdominal surgery: postanesthesia recovery characteristics. AB - BACKGROUND AND OBJECTIVES: Outcome studies comparing general anesthesia combined with epidural anesthesia (GEN-EPI) to general anesthesia (GEN) for major abdominal surgery have been equivocal. However, many believe that patients anesthetized with GEN-EPI fair better than GEN. This study tests the hypothesis that there are favorable recovery characteristics associated with GEN-EPI as compared with GEN following abdominal surgery. METHODS: A prospective randomized double-blind trial, consisting of 30 patients ages 18-74 undergoing abdominal surgery was undertaken. Patients received either GEN-EPI or GEN by standardized protocol. At the end of surgery the epidural catheter was removed and psychological testing was performed over 24 hours to determine recovery characteristics. These included the modified Slater test, the Self-Assessment Manikin, the Kendrick Digital Copying Test, the Hospital Anxiety and Depression Scale, as well as visual analog scales for pain and appearance. RESULTS: Patients receiving GEN-EPI emerged from anesthesia faster (P < .03) with less pain on awakening (P < .04 at rest; P < .01 on coughing), had better psychomotor function at 2 hours (P < .04), and were less drowsy at 4 hours (P < .04), than patients with GEN. There was no difference in pain intensity after the initial assessment, morphine usage, mood, anxiety, and depression at any other measurement period. CONCLUSION: Transient quantifiable differences in recovery characteristics exist between patients receiving GEN-EPI and GEN. PMID- 9338906 TI - Superficial and deep cervical plexus block for carotid artery surgery: a prospective study of 1000 blocks. AB - BACKGROUND AND OBJECTIVES: Cervical plexus blocks are performed for carotid surgery to allow neurological assessment of the awake patient. The aim of this study was to establish the frequency of success, complications, and patient acceptance of the technique. METHODS: One thousand superficial and deep cervical blocks were performed in 924 patients having carotid artery surgery. Data about the blocks were recorded prospectively and patients were followed up postoperatively by an independent anesthesiologist to assess patient acceptance of the technique. RESULTS: Lidocaine was the most frequently used anesthetic (88%). Surgical supplementation of the blocks was required in 53% of operations. Six blocks (0.6%) had clinical evidence of intravascular injection of local anesthetic. Sedation was required in 66% of operations and conversion to general anesthesia occurred in 25 (2.5%) of operations. Ninety-one percent of patients reported no problems with the block, and 93% stated that they would have the same anesthetic for any future similar surgery. CONCLUSIONS: We conclude that superficial and deep cervical plexus block has a high success rate, low complication rate, and high patient acceptance rate. Caution should, however, be exercised to ensure a low intravascular injection rate which is of most concern with this technique, because blood was aspirated in 30% of patients during performance of the block. PMID- 9338907 TI - Two cases of cauda equina syndrome following spinal-epidural anesthesia. AB - BACKGROUND AND OBJECTIVES: Cauda equina syndrome (CES) is a well-known complication of spinal and epidural anesthesia. Previous reports have implicated lidocaine, chloroprocaine, and procaine in its etiology, but not bupivacaine. METHODS: A 63-year-old man underwent transurethral resection of the prostrate for which he received bupivacaine with glucose intrathecally. Postoperative, he had difficulty in urination and defecation, and magnetic resonance imaging revealed spinal stenosis at the L1-L2 level. The second patient was a 70-year-old woman who underwent hip replacement surgery using a combined spinal-epidural technique. Postoperative, after 42 hours, when the epidural infusion of bupivacaine was stopped, the patient had difficulty in urination and defecation. No anatomical abnormality was found on magnetic resonance imaging. RESULTS: The two patients developed cauda equina syndrome following bupivacaine with glucose injected spinally, and bupivacaine without glucose injected in a combined spinal-epidural technique. CONCLUSIONS: This case report describes two cases of CES following the use of bupivacaine. The first patient had spinal stenosis which could explain this complication; however the explanation for CES in the second patient is uncertain and consequently speculative. We have discussed the possible contributing factors but believe that the etiology of CES in the second patient remains unknown. PMID- 9338908 TI - Comparative local anesthetic efficacy and pharmacokinetics of epidurally administered ropivacaine and bupivacaine in the sheep. AB - BACKGROUND AND OBJECTIVES: Ropivacaine is the S(-) propyl homolog of bupivacaine and mepivacaine. Studies in humans have confirmed the results of studies in laboratory animals that ropivacaine is a long-acting local anesthetic with an anesthetic profile similar to bupivacaine. Acute, intravenous systemic toxicity studies have been conducted in sheep and dogs. Local anesthetic efficacy has been studied after epidural administration in the dog. This study was initiated to determine the local anesthetic efficacy and pharmacokinetics of ropivacaine and bupivacaine after epidural administration in an experimental sheep model and to evaluate the sheep model as a model of experimental epidural anesthesia. METHODS: Twelve adult nonpregnant ewes were prepared with chronically implanted lumbar epidural catheters and arterial lines. Each sheep received injections of 5.0 mL ropivacaine and bupivacaine (0.5% and 0.75%) in a blinded, random, cross-over fashion. Onset and duration of sensory and motor blockade were evaluated. Arterial blood samples were drawn for serum drug concentration determinations and pharmacokinetic analysis. RESULTS: Onset and duration of motor blockade were similar for comparable concentrations of both drugs. Both concentrations of ropivacaine and bupivacaine 0.5% demonstrated differential neural blockade. The peak serum concentrations generally occurred within 8 minutes after administration. The terminal elimination half-life in serum was about 3.5-4.0 hours and 6 hours for ropivacaine and bupivacaine, respectively. No signs of systemic toxicity were observed. Results of sensory and motor blockade were consistent with previous studies in laboratory animals and humans. CONCLUSIONS: Ropivacaine produces sensory and motor blockade which is similar to that produced by equal concentrations of bupivacaine after epidural administration in the sheep. Peak serum concentrations produced no signs of systemic toxicity. The results of this study are consistent with previously published data from studies in laboratory animals and humans. The sheep model of experimental epidural anesthesia appears to be a clinically relevant method to evaluate experimental local anesthetics. PMID- 9338909 TI - Lidocaine does not depress reflex dilation of the pupil. AB - BACKGROUND AND OBJECTIVES: Pupillary dilation in response to dermatomal electrical stimulation is one method of determining sensory block level during combined epidural and general anesthesia. Use of this technique may, however, be confounded by systemic absorption of epidurally administered local anesthetics. Accordingly, the effects of intravenous lidocaine on the magnitude and duration of reflex pupillary dilation were evaluated. METHODS: Six volunteers were each anesthetized twice with desflurane 3.5-6.0%. During one anesthetic, intravenous lidocaine was administered to a plasma concentration of 5.3 +/- 1.5 micrograms/mL. When the plasma concentrations were stable, a 5-second tetanic electrical stimulus was applied. Pupil size was then recorded for 8 minutes. RESULTS: Lidocaine, at plasma concentrations near 5 micrograms/mL, did not significantly alter the pupillary response to electrical stimulation. In contrast, stimulus-induced increase in heart rate was obliterated. Painful stimulation did not increase systolic blood pressure in either case. CONCLUSIONS: Typical plasma lidocaine concentrations observed during epidural anesthesia are unlikely to prevent the use of pupillary responses to evaluate sensory block level. PMID- 9338910 TI - Trigger point injections for myofascial pain during epidural analgesia for labor. AB - BACKGROUND AND OBJECTIVES: Myofascial pain is the leading cause of chronic low back pain and in most cases can be successfully resolved with trigger point injections of local anesthetics. This type of pain can exist during pregnancy and exceed the analgesia provided by an epidural for labor. METHODS: A 31-year-old primiparous woman received an epidural infusion for labor analgesia. Despite complete resolution of labor pain and a solid, bilateral T10 block, the patient reported discomfort at two discrete locations in her right lumbar paraspinous muscle. RESULTS: The administration of local anesthetic via trigger point injections resulted in successful palliation of the myofascial pain. CONCLUSIONS: Myofascial pain can be an etiology of back pain in the parturient. Trigger point injections, even when used concomitantly with a functioning epidural infusion, can be a valuable aid for the provision of complete analgesia. PMID- 9338911 TI - Continuous parasacral sciatic nerve block: two case reports. AB - OBJECTIVE: This study investigated the use of a continuous parasacral sciatic nerve block for anesthesia and postoperative analgesia for lower extremity surgery. METHODS: A continuous parasacral sciatic nerve block was performed in two patients (triple ankle arthrodesis and below-knee amputation). The sacral plexus was identified using an insulated Tuohy needle and a nerve stimulator. A catheter was placed near the elements of the sacral plexus via the Tuohy needle. RESULTS: In both patients, surgical anesthesia was successfully established through the parasacral catheter with lidocaine 1% (1/200,000 epinephrine), and postoperative analgesia was successfully established with a bolus of bupivacaine 0.375% (1/200,000 epinephrine) and maintained with a continuous infusion of bupivacaine 0.1% (8 mL/h) for 48 hours. CONCLUSION: We conclude that continuous parasacral sciatic nerve block can provide anesthesia and long-term analgesia for operative procedures of the foot and leg. PMID- 9338912 TI - Experience with gabapentin for neuropathic pain in the head and neck: report of ten cases. AB - BACKGROUND AND OBJECTIVES: Gabapentin is an oral antiepileptic agent with an unknown mechanism of action. Recent case reports have suggested that gabapentin may be effective in the treatment of a variety of neuropathic pain states. This report presents baseline and follow-up data on ten patients who were treated with gabapentin when other pharmacologic interventions failed to relieve their neuropathic pain. METHODS: Ten patients referred for treatment of unrelieved neuropathic pain in the head and neck region were included in this study. Baseline and follow-up information included measures of pain intensity and pain quality. All of the patients were started on 300 mg gabapentin three times per day, though daily doses of up to 2400 mg were required for pain relief. RESULTS: Eight of the ten patients had no neuropathic pain on follow-up, whereas the remaining 2 patients reported only partial relief at follow-up. None of the patients complained of side effects. Gabapentin was effective in alleviating steady burning pain as well as lancinating pain and allodynia. CONCLUSIONS: The results suggest that gabapentin may be effective in the management of some cases of neuropathic pain in the head and neck. However, controlled, double-blind longitudinal studies are needed to evaluate this possibility. PMID- 9338913 TI - Unexplained neurologic deficit after uneventful combined spinal and epidural anesthesia for cesarean delivery. AB - BACKGROUND AND OBJECTIVES: Neurologic deficits after spinal and epidural anesthesia are uncommon and have a variety of pathophysiologic mechanisms. Local anesthetic neurotoxicity may occur, although subarachnoid bupivacaine has an unblemished clinical record. METHODS: A healthy parturient underwent uneventful combined spinal and epidural anesthesia for elective cesarean delivery. Intraspinal drugs administered included hyperbaric bupivacaine 0.5% and fentanyl (subarachnoid) and 2% lidocaine with epinepherine 1:200,000 and meperidine (epidural). RESULTS: On the third postpartum day the patient reported buttock numbness. An area of hypoesthesia in the distribution of the lower sacral nerves was present, without systemic or other neurologic symptoms and signs. The deficit persisted, and imaging of the lower spinal canal revealed no significant abnormality. Full recovery of sensation occurred after 7 months. CONCLUSION: Drug induced neurotoxicity is a possible explanation, although the exact etiology is uncertain. PMID- 9338914 TI - Propofol for opioid-induced side effects. PMID- 9338915 TI - Technique for sphenopalatine ganglion block. PMID- 9338916 TI - Does epidural morphine predispose to Guillain-Barre syndrome? PMID- 9338918 TI - Antinociceptive properties of adrenal chromaffin cells. PMID- 9338917 TI - Comment on case report of Kabeer and Hardy. PMID- 9338920 TI - Noxious stimuli should not be used to assess level of neural blockade. PMID- 9338919 TI - Knotting of a femoral catheter. PMID- 9338921 TI - Reply to Dr. Hogan regarding venous capacitance changes in the lower extremities during spinal anesthesia. PMID- 9338922 TI - Nutritional requirements for growth of an endophyte: Ceratopycnidium baccharidicola. AB - The carbon sources, nitrogen sources and vitamin requirements for the growth of Ceratopycnidium baccharidicola (an endophyte of Baccharis coridifolia) were studied. The fungus utilized several carbon sources: pectin, sucrose, fructose, glucose, maltose, cellobiose, xylose, arabinose, mannitol, mannose and sorbitol. Sucrose and fructose were found to be the best carbon sources. Nitrogen sources utilized by the endophyte included: nitrates, ammonium and amino acids such as proline, asparagine and glycine. Undefined complex nitrogen sources such as soytone, tryptone, yeast extract and casamino acids supported excellent growth. In a defined medium, thiamine was the only vitamin required for growth. Under optimum conditions the vegetative growth of C. baccharidicola was enhanced six fold over its growth in glucose-asparagine medium. PMID- 9338923 TI - Tendons and fluoroquinolones. Unresolved issues. PMID- 9338924 TI - The cost of osteoporosis in France. PMID- 9338925 TI - Multidisciplinary day hospital treatment of rheumatoid arthritis patients. Evaluation after two years. AB - The availability of multidisciplinary care for rheumatoid arthritis is still limited. The Raoul Dufy Program offered by the Saint-Antoine Teaching Hospital in Paris provides one-on-one personalized care in a day hospital setting as an adjunct to conventional medical follow-up. Listening and providing information and education are major objectives of the nurse, rheumatologist and physical therapist participating in the program. The team also includes a social worker, a surgeon, a dietician, a podiatrist and a psychologist, who intervene as needed. Seventy patients attended the program between December 1993 and September 1995 and were asked to complete a baseline and a three-month questionnaire designed to evaluate the effects of the program in terms of new therapeutic interventions, patient knowledge and quality of life. The patient knowledge score increased significantly (P < 0.0001). Many therapeutic interventions were initiated after program attendance, especially in the fields of podiatry, psychology and physical therapy. However, the quality of life score failed to improve. These results and the substantial patient demand for appointments are encouraging. Further work is needed on the methodology of multidisciplinary care evaluation. Coping strategy evaluation tools may allow to identify some of the specific benefits provided by the multidisciplinary approach. PMID- 9338926 TI - Evaluation of bone mineral density in patients with rheumatoid arthritis. Influence of disease activity and glucocorticoid therapy. AB - OBJECTIVES: To study bone mass and the factors that influence bone mass in rheumatoid arthritis patients versus controls. PATIENTS AND METHODS: 85 patients (73 women) with a mean age of 57 +/- 11 years and a mean disease duration of 13 +/- 9 years were compared to 85 age- and sex-matched controls. Among the patients, 62 (76%) had positive rheumatoid factor tests and 51 (60%) were receiving steroid therapy, with a mean daily dose of 10 +/- 4 mg and a mean duration of 7 +/- 6 years. The following parameters were determined: morning stiffness duration, painful and swollen joint counts, Lee's and Ritchie's indices, Health Assessment Questionnaire score, erythrocyte sedimentation rate, and C-reactive protein. Bone mineral density was measured at the lumbar spine and femoral neck using dual-energy X-ray absorptiometry (Sophos L-XRA). RESULTS: In the nonsteroid-treated patients, bone mineral density was similar to that in controls at the lumbar spine but was decreased by 8% (95% confidence interval [CI], 1.8-14.2%) at the femoral neck (0.76 +/- 0.14 g/cm2 versus 0.83 +/- 0.15 g/cm2; P = 0.03). Decreases of 11.5% (95% CI, 8.1-14.9%) at the lumbar spine and 10.4% (95% CI, 6.4-14.4%) at the femoral neck were found in the steroid-treated patients versus the nonsteroid-treated patients. In the patient group, femoral neck bone mineral density was significantly negatively correlated with age (r = 0.5), the Heath Assessment Questionnaire score (r = -0.27), and the erythrocyte sedimentation rate (r = -0.25), whereas only the first two variables were significantly correlated with lumbar bone mineral density. A multiple linear regression model including age, glucocorticoid use, rheumatoid factor, the Health Assessment Questionnaire score, and the erythrocyte sedimentation rate was constructed and adjusted for the number of variables. This model explained 44.7% of the variance of femoral neck bone mineral density. CONCLUSION: Rheumatoid arthritis is associated with a decrease in bone mass that is most marked in patients with active and/or severe disease and in those who take glucocorticoids. PMID- 9338927 TI - Validation of the European Spondylarthropathy Study Group and B. Amor criteria for spondylarthropathies in Lebanon. AB - OBJECTIVES: 1) To validate European Spondylarthropathy Study Group (ESSG) and B. Amor's criteria for spondylarthropathies in Lebanon. 2) To evaluate the frequency of spondylarthropathies in rheumatological practice in Lebanon. PATIENTS AND METHODS: Cases of definite and probable spondylarthropathy were diagnosed based on the clinical judgement of participating rheumatologists, without reference to the two criteria sets under study. The first two patients without spondylarthropathy seen after each spondylarthropathy case were included into the control group. Criteria in the ESSG and B. Amor sets were looked for in the patient and control groups. The frequency of spondylarthropathy meeting each criteria set was determined. RESULTS: Of the 841 patients evaluated during the study period, 68 met B. Amor's criteria and 72 met ESSG criteria. There were 29 cases of ankylosing spondylitis (40.3%), ten of peripheral psoriatic arthritis (13.8%), two of reactive arthritis (2.8%), two of enteropathic arthropathy (2.8%), and 29 of undifferentiated spondylarthropathy (40.3%). In the definite spondylarthropathy group, sensitivity and specificity were 77.19% and 97.55% for B. Amor's criteria versus 91.23% and 100% for ESSG criteria. The frequency of spondylarthropathy was 8.1% (95% confidence interval [CI], 6.3-9.9) or 8.56% (CI 6.6-10.5) according to B. Amor and ESSG criteria, respectively. CONCLUSION: Our data validate both criteria sets in the Lebanese population, demonstrating that they are useful in populations that are genetically different from the European populations used to develop them. Spondyloarthropathy is the most common in our rheumatology practice. PMID- 9338928 TI - Magnetic resonance imaging and axial involvement in spondylarthropathies. Delineation of the spinal entheses. AB - We used magnetic resonance imaging in a prospective cross-sectional study to evaluate the components of axial involvement in spondylarthropathies, to determine whether the entire intervertebral disk is an enthesis and to gauge how useful this imaging technique is in detecting enthesitis of the spine. Thirty-one patients with spondylarthropathies and 14 controls with mechanical spinal disease were included. Images of the thoracic and lumbar spine were obtained using plain radiography, radionuclide bone scanning, and magnetic resonance imaging (sagittal sections, T1-weighted sequences before and after gadolinium injection and fat saturation and T2-weighted sequences). Magnetic resonance imaging signal abnormalities reflected inflammation and hypervascularization of the subchondral bone underlying the affected entheses (low signal enhancing after gadolinium and fat saturation on T1 images, high signal on T2 images). These abnormalities were often visible early in the disease process, at a time when there were not yet any clinical manifestations or radiographic or bone scan changes. In addition to showing involvement of the classic spinal entheses, magnetic resonance imaging also demonstrated evidence of inflammation and hypervascularization of the central part of the vertebral endplates and intervertebral disks, confirming that the center of the disk is an enthesis and that inflammatory enthesitis is the mechanism underlying at least some cases of discitis seen in patients with spondylarthropathies. PMID- 9338929 TI - Do minocycline and other tetracyclines have a place in rheumatology? AB - Tetracyclines are a family of antimicrobials with activity against a broad range of organisms including those that develop intracellularly. Links have been reported between some infections and some inflammatory joint diseases, with the most notable example involving mycoplasmas and rheumatoid arthritis. Reactive arthritides are known to be triggered by organisms found in the gastrointestinal or genitourinary tract, and antigenic material from these organisms has recently been demonstrated in synovial tissue from patients with reactive arthritis. These facts led to the hypothesis that tetracyclines may be useful in rheumatoid arthritis and reactive arthritis. Two controlled studies found that minocycline benefited rheumatoid arthritis patients when it was given either as an adjunct to another second-line treatment or as the only slow-acting drug. Lymecycline has been found to expedite recovery from reactive arthritis due to Chlamydia trachomatis, and tetracycline to decrease the incidence of reactive arthritis due to sexually-transmitted diseases. The safety profiles of these treatments were acceptable in all available studies but require further investigation during long term administration. The benefits may be related to the immunomodulating effects of tetracyclines and/or to their ability to inhibit metalloproteases such as collagenases. Whether tetracycline therapy influences the course of radiologic lesions in rheumatoid arthritis remains unknown. However, minocycline therapy has given sufficient proof of its efficacy to make it an attractive alternative in rheumatoid arthritis. PMID- 9338930 TI - Algodystrophy (reflex sympathetic dystrophy syndrome) and causalgia: novel concepts regarding the nosology, pathophysiology, and pathogenesis of complex regional pain syndromes. Is the sympathetic hyperactivity hypothesis wrong? AB - Concepts regarding the nosology, pathophysiology and pathogenesis of reflex sympathetic dystrophy syndrome are currently in a state of flux. Causalgia and reflex sympathetic dystrophy syndrome are now generally felt to be on the same continuum and as a result interest for defining criteria for the latter condition has waned. The pathogenic role of adrenergic sympathetic activity has been so successfully challenged that the last international consensus conference judged inappropriate any reference to the sympathetic system in the terms used to designate these conditions, thus confirming the position long defended by most French authors. The vasomotor abnormalities may be due to antidromic release of neuromediators by the endings of polymodal C fibers. These fibers do not belong to the sympathetic system but often travel with sympathetic nerves, a characteristic that may explain the efficacy of sympathetic nerve blocks, although other possibilities exist including a placebo effect. Also, efferent sympathetic fibers may undergo activation by nonadrenergic mediators. The mechanisms capable of initiating and perseverating activation of polymodal C afferents are being actively investigated and have been found to exhibit similarities with the mechanisms underlying peripheral and central sensitization of pain-producing afferents. Growth factors, such as nerve growth factor, may play an important role in causalgia. In "reflex sympathetic dystrophy syndrome", microcirculatory stasis may contribute to the initiation or perpetuatation of the disorders. Further work on the nerve supply to the venular network and on the venoarterial reflex is needed. PMID- 9338931 TI - Spinal neurenteric cyst. A report of a case. AB - A case of intradural extramedullary neurenteric cyst is reported. The embryogenesis and surgical treatment of this lesion are discussed. PMID- 9338933 TI - Giant cell arteritis involving the lower limbs. AB - Giant cell arteritis is an inflammatory disease that can affect the arteries anywhere in the body. Two cases are reported in which the arteries of the lower limbs were involved. Intermittent claudication with a walking distance of only 30 m was the inaugural manifestation in both cases. A biopsy of the superficial femoral artery provided the diagnosis in the first case. Ergotamine toxicity was considered initially in the second case. Acute ischemia and gangrene requiring amputation can complicate giant cell arteritis of the lower limbs and consequently corticosteroid therapy in an effective dose should be given as soon as the diagnosis is made. The inflammatory arterial lesions improve under therapy, but irreversible fibrosis with stenosis can develop if treatment is initiated late. PMID- 9338932 TI - Cauda equina syndrome with pagetic vertebral fusion. Clinical recovery under calcium-vitamin D supplementation plus clodronate after apparent failure of pamidronate and acquired resistance to etidronate. AB - A patient with an osteolytic L2-L3 pagetic block and pagetic lesions of L1 and the sacrum seen only as increased radionuclide activity became resistant to etidronate after the fifth course (5 mg/kg/d six months per year) and developed severe cauda equina syndrome (reduction in walking distance to 30 m and sphincter dysfunction) due primarily to vertebral hypertrophy. Five months after a ten-day course of intravenous pamidronate (22.5 mg/d), the clinical symptoms were unchanged, although the alkaline phosphatase level was down 50%. Oral clodronate (1,600 mg/day for six months per year) in combination with calcium and vitamin D supplementation dramatically improved the walking distance and sphincter disorders. Resolution of the neurological manifestations was complete after the second clodronate course. At last follow-up nine months after the fourth clodronate course, there was no evidence of a relapse and the alkaline phosphatase level was normal. The time course of events in this patient does not allow to affirm that pamidronate was ineffective and suggests that calcium and vitamin D supplementation improved mineralization of the pagetic block and enhanced the effect of bisphosphonate therapy. PMID- 9338934 TI - Rapidly destructive disk degeneration. Report of a case documented by computed tomography and magnetic resonance imaging. AB - A case of rapidly destructive disk degeneration in which imaging studies were performed over a two-year period is reported. Rapidly destructive disk degeneration is a clinicopathological entity recently redefined by Revel et al.. It is analogous to conditions characterized by peripheral chondrolysis, such as rapidly destructive hip osteoarthritis. Its pathogenesis remains obscure. PMID- 9338935 TI - Acute sacroiliitis as a manifestation of calcium pyrophosphate dihydrate crystal deposition disease. A report of two cases. AB - Whereas radiographic lesions of the sacroiliac joints are common in patients with calcium pyrophosphate deposition crystal disease, they are rarely accompanied with clinical symptoms. We report two cases of acute sacroiliitis probably due to calcium pyrophosphate dihydrate deposition disease. The patients were a 53-year old man and an 82-year-old woman with chondrocalcinosis in other joints and presence on computed tomography studies of the sacroiliac joints of sclerosis and irregularities of the joint margins with a thin linear calcific deposit within the joint. Both patients recovered fully under therapy with colchicine, analgesics and rest. These two cases suggest that acute sacroiliitis can be caused by calcium pyrophosphate dihydrate crystal deposition disease. PMID- 9338936 TI - Cerebral vein thrombosis after an intrathecal glucocorticoid injection. AB - We report two cases of cerebral venous thrombosis that occurred after an intrathecal glucocorticoid injection for common lumbosciatic syndrome. The patients were two females, aged 34 and 47 years, who developed typical post lumbar puncture headache. The manifestations leading to the diagnosis of cerebral venous thrombosis were status epilepticus in one case and persistent headache in the other. Both patients were using oral contraceptives. Grynblat et al. (Rev Rhum [Engl Ed] 1995; 62: 691) recently reported two cases in young women of cerebral venous thrombosis after intrathecal administration of a glucocorticoid or an iodinated contrast agent. These cases are useful reminders that intrathecal glucocorticoid injections can be followed by cerebral venous thrombosis, and that younger women may be at increased risk for this complication. PMID- 9338937 TI - Prevalence of sacroiliac involvement in systemic lupus erythematosus. A retrospective study of forty-one patients. PMID- 9338938 TI - Myeloma and thymoma: an as yet unreported association. PMID- 9338939 TI - Extensive reflex sympathetic dystrophy syndrome of the lower limbs leading to a diagnosis of osteomalacia due to Fanconi syndrome. PMID- 9338940 TI - Septic arthritis of the sternoclavicular joint with an unusual portal of entry. PMID- 9338943 TI - Skeletal effects of menstrual disturbances in athletes. AB - This article reviews the skeletal effects and clinical implications of menstrual disturbances in active women. At the lumbar spine, menstrual disturbances are associated with premature bone loss or failure to reach peak bone mass, while appendicular sites are less affected. This suggests that trabecular bone is more sensitive to hormonal stimuli and less responsive to mechanical loading than cortical bone. Although the mechanisms responsible for the detrimental effects of menstrual disturbances are likely to be multifactorial, low circulating levels of oestrogen are thought to be the main cause. The clinical significance of menstrual disturbances depends upon a number of factors, including type of sport, genetic background, body composition and calcium intake. Not all athletes who present with menstrual disturbances will develop osteopenia. Nevertheless, the risk of stress fracture does seem to be increased in athletes with menstrual disturbances and with lower bone density. Whether athletes with menstrual disturbances are at a greater risk for osteoporosis in later life is not yet known. Bone loss can be at least partially reversed, especially with the spontaneous resumption of menses. This may serve to offset any previous increased risk of osteoporosis. Furthermore, other factors, apart from low bone mass, act to determine the likelihood of osteoporotic fractures. Therefore, the clinical significance of menstrual disturbances associated with exercise participation needs to be established for each individual athlete. Bone densitometry may guide the clinician in this respect and assist in the formulation of appropriate management strategies. PMID- 9338941 TI - Safety of combination therapy with hydroxychloroquine, gold sodium thiomalate and methotrexate in early rheumatoid arthritis. PMID- 9338942 TI - Anti-doping control against scientific fraud: an urgent quality assurance measure. PMID- 9338944 TI - Pronounced resting bradycardia in male elite runners is associated with high heart rate variability. AB - Forty-eight hour Holter monitoring was undertaken of 16 male elite middle- and long-distance runners, age 25 +/- 3 years, with peak oxygen uptake 4.83 +/- 0.43 1 O2/min or 73.0 +/- 3.9 ml O2/kg/min. The athletes had pronounced bradycardia during the night-time, with heart rate calculated from four RR intervals < 30 beats/min in five runners. Twelve of 16 runners had RR intervals > 2 s. Of those, 10 runners had sinus pauses exceeding 2 s, the longest being 3.06 s. Three runners had AV block II, two with Mobitz type 1, and one with both Mobitz type 1 and 2. Autonomic function was estimated by time domain and power spectral analysis of heart rate variability. The runners were compared with a control group of 13 sedentary or moderately active subjects. The runners had a mean of 14 b.p.m. lower heart rate at night than the controls. The runners had higher heart rate variability in all spectral bands. In the time domain pNN50 and rMSSD, which are considered to reflect strongly vagal tone, were markedly higher in the runners than the controls. The findings suggest that an increased parasympathetic tone might at least partly explain the pronounced resting sinus bradycardias found in endurance-trained runners. PMID- 9338945 TI - Passive knee extension test to measure hamstring muscle tightness. AB - The purpose of this study was twofold: (a) to examine the reliability of a test designed to measure tightness of the hamstring muscles, and (b) to assess the pelvic motion during this test. The knee was passively extended by a standardized force, while the hip was stabilized in 120 degrees of flexion. The knee angle was measured with a goniometer and represents the hamstring tightness. Twenty-eight test-retests were performed. The correlation coefficient was found to be 0.99, and the CV was found to be 1%. We used a MacReflex measurement system to assess the associated pelvic motion. Eight measurements were taken, and the median of associated pelvic motion was 4.1 degrees. It is concluded that the passive knee extension test is a simple and reliable method, and the associated pelvic motion is minimal. PMID- 9338946 TI - Development of physical response after athletics training in adolescents with Down's syndrome. AB - Starting in January 1994, a group of 20 adolescents with Down's syndrome began a competition-oriented athletics training program at a public sports stadium twice a week as part of their education towards integration. They were given complete medical examinations beforehand and followed medically and professionally throughout the program. The results showed a significant improvement in the test scores measuring strength, speed and endurance and a tendency towards an athletic morphotype. PMID- 9338947 TI - Registration of cruciate ligament injuries in Norwegian top level team handball. A prospective study covering two seasons. AB - All cruciate ligament injuries in the three upper divisions for men and women (3392 players) in Norwegian team handball in the 1989-90 and 1990-91 seasons were registered. A questionnaire was mailed to all injured players. Ninety-three cruciate ligament injuries were registered; 87 in the anterior cruciate ligament (ACL), and six in the posterior cruciate ligament (PCL). Among women, 1.8% were injured compared with 1.0% of the men. In the first division, the risk of being injured was considerably higher: 4.5% of the players had a cruciate ligament injury. There were 0.97 cruciate ligament injuries per 100 playing hours in the three divisions taken together. Seventy-five per cent of the injuries occurred during games. Ninety-five per cent involved no contact between players. Activities in which the friction between shoe and floor was significant caused 55% of the injuries. Injuries caused by running into another player contributed to only 5% of the injuries. No significant differences were observed in injury incidence during matches between different types of floors (parquet, Pulastic and other synthetic surfaces.) PMID- 9338948 TI - Fitness training and its effect on musculoskeletal pain in professional ballet dancers. AB - In a controlled, prospective, randomized study, half of the dancers in a professional ballet company were asked to do extra self-administered fitness training, while the other half became the control group. The aim was to examine if the dancers in the training group would be able to keep up the extra training during a regular season and to examine its effect on their maximum oxygen uptake and on their self-estimated musculoskeletal pain. The training group increased their oxygen uptake more than the control group. The self-estimated functional inability because of pain (SEFIP) indicated significantly less pain the week after the premiere for the study population taken as a whole, but not for the two groups when considered separately. The training group claimed that the fitness training had helped them to cope with the psychological strain during rehearsals. PMID- 9338949 TI - Long-term results after surgical management of partial Achilles tendon ruptures. AB - Although Achilles tendon injuries are common overuse injuries in sports, the exact incidence is unknown, primarily as a result of varying definitions and diagnoses of the underlying pathological changes. Despite numerous studies of treatment of the Achilles tendon injuries, the long-term results are not well known. The results after surgical treatment of chronic partial Achilles tendon ruptures in 64 patients with a follow-up of 6 (1.5-11) years were evaluated in a retrospective study. The ruptures were divided into three groups: (I) proximal (more than 3 cm above the calcaneus), (II) distal and (III) combined (proximal and distal). All patients underwent an operation involving the excision of the devitalized tendon tissue and, in groups (II) and (III), also the excision of the deep Achilles bursa and removal of the dorsal corner of the calcaneus. The functional results were satisfactory in 43 (67%) patients and unsatisfactory in 21 (33%). The results were better in patients with proximal ruptures than in patients with either distal or combined ruptures. Males experienced better results than females. Post-operative immobilization in a plaster cast had no significant influence on the final result. Nine (14%) patients with either a distal or a combined rupture were re-operated on and in seven of them the final result was satisfactory. The conclusion of this study is that partial Achilles tendon ruptures are often difficult to treat and only two out of three patients can be expected to obtain satisfactory results after surgical treatment. PMID- 9338950 TI - Injury proneness in infantry conscripts undergoing a physical training programme: smokeless tobacco use, higher age, and low levels of physical fitness are risk factors. AB - Musculoskeletal injuries represent an adverse event of strenuous physical activity. The aim of the present study was to identify pretraining factors that may predispose to such injuries. Risks of injury according to age, body composition, previous physical activity, physical fitness, use of smokeless tobacco (moist snuff) and smoking habits were determined in a population of 480 male conscripts in the army. Data were obtained by questionnaire, height and weight measurements, and from a 3000-metre run test prior to a 10-week period of basic military and physical training. Injuries were registered by doctors attached to the training camp. Every fourth conscript sustained one or more musculoskeletal injuries during the training period. Low back pain, overuse knee injuries, Achilles tendinitis, and sprains of joint capsules or ligaments were the most frequent diagnosis groups. Subjects aged 22 years and more, the least active persons before call-up, those who thought they were less fit than the average, the slowest one-third in the 3000-metre run test, smokers of more than 10 cigarettes a day, and snuff-takers suffered more injuries according to univariate analyses. Multiple logistic regression analysis showed that age, self assessed physical fitness and snuff-taking were mutually independent risk factors of high statistical significance. PMID- 9338951 TI - Exercise and diet interventions improve perceptions of self in middle-aged adults. AB - The effect of a 1-year exercise and diet intervention program on global self concept, perceptions of the body, physical competence, exercise mastery, social competence, social comfort, and fitness was examined with 208 healthy individuals (191 males, 17 females) aged 39-49 years (mean age 44.9) with elevated risk factors for cardiovascular disease. The relative utility of the skill development versus the self-enhancement model of the self-concept/behaviour relationship was tested. The participants were randomized into four groups: diet (n = 53), diet and exercise (n = 64), exercise (n = 48) and no active intervention (n = 43). Measurements were made by the Harter adult self perception profile (HASPP) and the self-perception in exercise questionnaire (SPEQ). Two-way ANOVA analyses revealed that exercise participation, with or without diet, enhanced self perceptions of physical mastery and ability, body perception, fitness and social comfort. The unique contribution of diet indicated enhanced body perception. No effect was found of diet or exercise on global self-concept. Exercise participation had a positive effect on perceptions of the self, and the higher the compliance with the exercise program, the stronger were the effects on the self-perceptions. This supported the skill development model of the self concept/behaviour relationship. As the pretest self-concept scores did not predict exercise compliance, the self-enhancement model of the self concept/behaviour relationship was not supported. PMID- 9338952 TI - Introduction to the special issue: mental health prevention science in rural communities and contexts. PMID- 9338953 TI - Challenges in defining and developing the field of rural mental disorder preventive intervention research. AB - An overview of selected issues and challenges in defining and developing the field of rural preventive intervention research is presented. One fundamental challenge is to clarify the distinguishing characteristics of prevention science in rural contexts. Other challenges are evident in the need to address: the lack of consensus on conceptual and methodological approaches to this field, limited empirical study to date, the tremendous diversity of rural populations, and inconsistencies in the usage of the term "rural". This article suggests the organization of a work group to formulate and implement a clear research agenda. In addition, several general questions are discussed that, if addressed, might serve to better define and further develop the field. These questions concern the implications of multiple approaches to prevention science in rural contexts, the classification of rural populations, the functional relevance of rural residence in the etiology of specific disorders, the application of extant etiological models to interventions designed specifically for rural populations, the conduct of rural area prevention needs assessments, the development of models for collaboration between intervention researchers and rural community stakeholders, and strategies to engage rural residents in preventive interventions. PMID- 9338954 TI - Psychological distress and help seeking in rural America. AB - The implications of exposure to acute and chronic stressors, and seeking mental health care, for increased psychological distress are examined. Research on economic stress, psychological distress, and rural agrarian values each point to increasing variability within rural areas. Using data from a panel study of 1,487 adults, a model predicting changes in depressive symptoms was specified and tested. Results show effects by size of place for men but not for women. Men living in rural villages of under 2,500 or in small towns of 2,500 to 9,999 people had significantly greater increases in depressive symptoms than men living in the country or in larger towns or cities. Size of place was also related to level of stigma toward mental health care. Persons living in the most rural environments were more likely to hold stigmatized attitudes toward mental health care and these views were strongly predictive of willingness to seek care. The combination of increased risk and less willingness to seek assistance places men living in small towns and villages in particular jeopardy for continuing problems involving depressed mood. PMID- 9338955 TI - Conducting ecologically valid prevention research: recruiting and retaining a "whole village" in multimethod, multiagent studies. AB - Many prevention studies are now designed with complementary interventions in different settings. Evaluations of these interventions require assessing the child's behavior in each of these settings. Conducting these studies, therefore, may involve recruiting school districts, principals, classroom teachers, peers, parents, siblings, and in later years, employers and intimate partners. These participants may be considered natural raters or satellite subjects, depending on their degree of involvement. Issues of recruitment and retention thus are magnified in multimethod, multiagent studies. To illustrate these issues, findings are presented for three studies conducted with risk populations in the past decade at the Oregon Social Learning Center: a passive longitudinal study, a selected prevention study, and an indicated prevention study. Findings indicate that achieving high recruitment and retention rates for at-risk and high-risk subjects in multisetting studies is possible, and that a developmental approach should be taken to recruiting risk populations. PMID- 9338956 TI - Implementing a comprehensive program for the prevention of conduct problems in rural communities: the Fast Track experience. The Conduct Problems Prevention Research Group. AB - Childhood conduct problems are predictive of a number of serious long-term difficulties (e.g., school failure, delinquent behavior, and mental health problems), making the design of effective prevention programs a priority. The Fast Track Program is a demonstration project currently underway in four demographically diverse areas of the United States, testing the feasibility and effectiveness of a comprehensive, multicomponent prevention program targeting children at risk for conduct disorders. This paper describes some lessons learned about the implementation of this program in a rural area. Although there are many areas of commonality in terms of program needs, program design, and implementation issues in rural and urban sites, rural areas differ from urban areas along the dimensions of geographical dispersion and regionalism, and community stability and insularity. Rural programs must cover a broad geographical area and must be sensitive to the multiple, small and regional communities that constitute their service area. Small schools, homogeneous populations, traditional values, limited recreational, educational and mental health services, and politically conservative climates are all more likely to emerge as characteristics of rural rather than urban sites (Sherman, 1992). These characteristics may both pose particular challenges to the implementation of prevention programs in rural areas, as well as offer particular benefits. Three aspects of program implementation are described in detail: (a) community entry and program initiation in rural areas, (b) the adaptation of program components and service delivery to meet the needs of rural families and schools, and (c) issues in administrative organization of a broadly dispersed tricounty rural prevention program. PMID- 9338957 TI - The extension service as key mechanism for research and services delivery for prevention of mental health disorders in rural areas. AB - The extension service associated with each state's land grant institution is an important resource for both programming and conducting research for the prevention of mental health disorders. This paper briefly outlines the history and structure of the extention service and gives examples of relevant programming across the country related to mental health. Although this programming is relevant to both rural and urban settings, this paper focuses on prevention programs and research in rural areas. Next is a description of extention's potential contributions to the prevention research process at each step of the Institute of Medicine's Preventive Intervention Research Cycle, followed by an illustration of an effective partnership between a research team and extension. Some of the ways in which extension can assist in the research process include doing local needs assessment and determining prevalence rates of a particular problem, reviewing literature on risk and protective factors, providing input on particular communities' needs and characteristics, and becoming a trusted link between local citizens and the research community. The article concludes with barriers to such an effective partnership and ways of reducing those hindrances. PMID- 9338958 TI - Prevention research in rural communities: overview and concluding comments. AB - This paper provides an overview of the challenges that confront researchers in rural settings, synthesizing the manuscripts in this special issue of The American Journal of Community Psychology. Researchers typically focus on issues of research design, measurement, and data analyses. However, when applied research is conducted in rural settings, greater time and attention are required to identify how the research can be conducted successfully. In this overview of the challenges that confront researchers in rural contexts, qualitative differences between rural and urban environments are described with particular attention to their implications for the conduct of rural research. Finally, theoretical and research topics that can better inform future rural research efforts are discussed. PMID- 9338959 TI - A novel cell ablation strategy blocks tobacco anther dehiscence. AB - We utilized a new cell ablation strategy to ablate specific anther cell types involved in the dehiscence process. The tobacco TA56 gene promoter is active within the circular cell cluster, stomium, and connective regions of the anther at different developmental stages. We introduced a cytotoxic TA56/barnase gene into tobacco plants together with three different anticytotoxic barstar genes. The anticytotoxic barstar genes were used to protect subsets of anther cell types from the cytotoxic effects of the TA56/barnase gene. The chimeric barstar genes were fused with (1) the tobacco TP12 gene promoter that is active at high levels in most anther cell types; (2) the soybean lectin gene promoter that is active earlier in the connective, and at lower levels in the circular cell cluster and stomium, than is the TA56 promoter; and (3) the tobacco TA20 gene promoter that is active at high levels in most anther cell types but has a different developmental profile than does the TP12 promoter. Normal anther development and dehiscence occurred in plants containing the TA56/barnase and TP12/barstar genes, indicating that barstar protects diverse anther cell types from the cytotoxic effects of barnase. Anthers containing the TA56/barnase and lectin/barstar genes also developed normally but failed to dehisce because of extensive ablation of the circular cell cluster, stomium, and contiguous connective regions. Anthers containing the TA56/barnase and TA20/barstar genes failed to dehisce as well. However, only the stomium region was ablated in these anthers. The connective, circular cell cluster, and adjacent wall regions were protected from ablation by the formation of barnase/barstar complexes. We conclude that anther dehiscence at flower opening depends on the presence of a functional stomium region and that chimeric barnase and barstar genes containing promoters that are active in several overlapping cell types can be used for targeted cell ablation experiments. PMID- 9338961 TI - The Ustilago maydis regulatory subunit of a cAMP-dependent protein kinase is required for gall formation in maize. AB - In the plant, filamentous growth is required for pathogenicity of the corn smut pathogen Ustilago maydis. Earlier, we identified a role for the cAMP signal transduction pathway in the switch between budding and filamentous growth for this fungus. A gene designated ubc1 (for Ustilago bypass of cyclase) was found to be required for filamentous growth and to encode the regulatory subunit of a cAMP dependent protein kinase (PKA). Here, we show that ubc1 is important for the virulence of the pathogen. Specifically, ubc1 mutants are able to colonize maize plants and, like the wild-type pathogen, cause localized symptoms in association with the presence of hyphae. However, in contrast to plants infected with wild type cells that often developed galls from initially chlorotic tissue, plants infected with the ubc1 mutant did not produce galls. These data suggest that PKA regulation is critical for the transition from saprophytic to pathogenic growth and from vegetative to reproductive development. Plate mating assays in which exogenous cAMP was applied suggested that the cAMP and b mating-type morphogenetic pathways may be coordinated. PMID- 9338960 TI - The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. AB - The cpr5 mutant was identified from a screen for constitutive expression of systemic acquired resistance (SAR). This single recessive mutation also leads to spontaneous expression of chlorotic lesions and reduced trichome development. The cpr5 plants were found to be constitutively resistant to two virulent pathogens, Pseudomonas syringae pv maculicola ES4326 and Peronospora parasitica Noco2; to have endogenous expression of the pathogenesis-related gene 1 (PR-1); and to have an elevated level of salicylic acid (SA). Lines homozygous for cpr5 and either the SA-degrading bacterial gene nahG or the SA-insensitive mutation npr1 do not express PR-1 or exhibit resistance to P. s. maculicola ES4326. Therefore, we conclude that cpr5 acts upstream of SA in inducing SAR. However, the cpr5 npr1 plants retained heightened resistance to P. parasitica Noco2 and elevated expression of the defensin gene PDF1.2, implying that NPR1-independent resistance signaling also occurs. We conclude that the cpr5 mutation leads to constitutive expression of both an NPR1-dependent and an NPR1-independent SAR pathway. Identification of this mutation indicates that these pathways are connected in early signal transduction steps and that they have overlapping functions in providing resistance. PMID- 9338962 TI - A conserved family of WD-40 proteins binds to the retinoblastoma protein in both plants and animals. AB - In mammalian cells, the retinoblastoma (RB) protein regulates G1 progression and functions through its association with various cellular proteins. Two closely related mammalian RB binding proteins, RbAp48 and RbAp46, share sequence homology with the Msi1 protein of yeast. MSI1 is a multicopy suppressor of a mutation in the IRA1 gene involved in the Ras-cAMP pathway that regulates cellular growth. Human RbAp48 is present in protein complexes involved in histone acetylation and chromatin assembly. We report the cloning of cDNAs encoding four plant RbAp48- and Msi1-like proteins: one from tomato, LeMSI1, and three from Arabidopsis. Complementation studies confirm that LeMSI1 can function as a multicopy suppressor of the yeast ira1 mutant phenotype. The LeMSI1 protein localizes to the nucleus and binds to a 65-kD protein in wild-type as well as ripening inhibitor (rin) and Neverripe (Nr) tomato fruit. LeMSI1 also binds to the human RB protein and the RB-like RRB1 protein from maize, indicating that this interaction is conserved between plants and animals. PMID- 9338963 TI - PCF1 and PCF2 specifically bind to cis elements in the rice proliferating cell nuclear antigen gene. AB - We have previously defined the promoter elements, sites IIa and IIb, in the rice proliferating cell nuclear antigen (PCNA) gene that are essential for meristematic tissue-specific expression. In this study, we isolated and characterized cDNAs encoding proteins that specifically bind to sites IIa and IIb. The two DNA binding proteins, designated PCF1 and PCF2, share > 70% homology in common conserved sequences at the N-terminal regions. The conserved regions are responsible for DNA binding and homodimer and heterodimer formation, and they contain a putative basic helix-loop-helix (bHLH) motif. The structure and DNA binding specificity of the bHLH motif are distinguishable from those of other known bHLH proteins that bind to the E-box. The motif is > 70% homologous to several expressed sequence tags from Arabidopsis and rice, suggesting that PCF1 and PCF2 are members of a novel family of proteins that are conserved in monocotyledons and dicotyledons. A supershift assay using an anti-PCF2 antibody showed the involvement of PCF2 in site IIa (site IIb) binding activities in rice nuclear extracts, particularly in meristematic tissues. PCF1 and PCF2 were also more likely to form heterodimers than homodimers. Our results suggest that PCF1 and PCF2 are involved in meristematic tissue-specific expression of the rice PCNA gene through binding to sites IIa and IIb and formation of homodimers or heterodimers. PMID- 9338964 TI - Characterization of oleosins in the pollen coat of Brassica oleracea. AB - Mature Brassica oleracea pollen grains are covered with a lipophilic pollen coat containing a variety of proteins. Screening of an anther cDNA expression library for the coding sequences of such proteins resulted in the isolation of a number of cDNA clones encoding glycine-rich oleosins. The proteins were shown to be attached to the lipophilic coat material only and to be absent elsewhere in the plant. Within the coat, several forms of the pollen coat oleosin with different molecular weights were detected. The forms are encoded by different transcripts that originate from a single gene. Expression of this gene is restricted to the tapetum and is quantitatively regulated by the water content of the anther. Similar oleosins were found in the pollen coat of B. alboglobra and B. napus. PMID- 9338965 TI - Recombination occurs uniformly within the bronze gene, a meiotic recombination hotspot in the maize genome. AB - The bronze (bz) gene is a recombinational hotspot in the maize genome: its level of meiotic recombination per unit of physical length is > 100-fold higher than the genome's average and is the highest of any plant gene analyzed to date. Here, we examine whether recombination is also unevenly distributed within the bz gene. In yeast genes, recombination (conversion) is polarized, being higher at the end of the gene where recombination is presumably initiated. We have analyzed products of meiotic recombination between heteroallelic pairs of bz mutations in both the presence and absence of heterologies and have sequenced the recombination junction in 130 such Bz intragenic recombinants. We have found that in the absence of heterologies, recombination is proportional to physical distance across the bz gene. The simplest interpretation for this lack of polarity is that recombination is initiated randomly within the gene. Insertion mutations affect the frequency and distribution of intragenic recombination events at bz, creating hotspots and coldspots. Single base pair heterologies also affect recombination, with fewer recombination events than expected by chance occurring in regions of the bz gene with a high density of heterologies. We also provide evidence that meiotic recombination in maize is conservative, that is, it does not introduce changes, and that meiotic conversion tracts are continuous and similar in size to those in yeast. PMID- 9338967 TI - Expression of expansin genes is correlated with growth in deepwater rice. AB - Expansins are a family of proteins that catalyze long-term extension of isolated cell walls. Previously, two expansin proteins have been isolated from internodes of deepwater rice, and three rice expansin genes, Os-EXP1, Os-EXP2, and Os-EXP3, have been identified. We report here on the identification of a fourth rice expansin gene, Os-EXP4, and on the expression pattern of the rice expansin gene family in deepwater rice. Rice expansin genes show organ-specific differential expression in the coleoptile, root, leaf, and internode. In these organs, there is increased expression of Os-EXP1, Os-EXP3, and Os-EXP4 in developmental regions where elongation occurs. This pattern of gene expression is also correlated with acid-induced in vitro cell wall extensibility. Submergence and treatment with gibberellin, both of which promote rapid internodal elongation, induced accumulation of Os-EXP4 mRNA before the rate of growth started to increase. Our results indicate that the expression of expansin genes in deepwater rice is differentially regulated by developmental, hormonal, and environmental signals and is correlated with cell elongation. PMID- 9338968 TI - A novel protein with DNA binding activity from tobacco chloroplast nucleoids. AB - A 41-kD DNA binding protein with a basic pl was purified from chloroplast nucleoids in photomixotrophically cultured tobacco cells, and its amino acid sequence was determined. Using this sequence information, its cDNA (CND41) was isolated, and its nucleotide sequence was determined. The predicted amino acid sequence of CND41 has a transit peptide of 120 amino acids and a mature protein of 382 amino acids. A distinctive helix-turn-helix motif in the lysine-rich N terminal region of the mature protein and an aspartyl protease active site motif were predicted. Expression of a series of truncated CND41 proteins in Escherichia coli indicated that the lysine-rich region is essential for DNA binding and that CND41 nonspecifically binds chloroplast DNA. Protein gel blot analyses showed CND41 mainly in cells and/or tissues containing nonphotosynthesizing, actively growing plastids. In addition, the accumulation of chloroplast transcripts in these cells and/or tissues (e.g., transcripts for QB binding protein of photosystem II [psbA] and large subunit of ribulose bisphosphate carboxylase [rbcL]) was negatively correlated with the accumulation of CND41. Analyses of cultured cells of transgenic tobacco with reduced CND41 levels showed a higher level of expression of chloroplast genes compared with that of the wild type. We discuss the possible function of CND41 as a negative regulator of chloroplast gene expression. PMID- 9338969 TI - Coexpression of the maize delta-zein and beta-zein genes results in stable accumulation of delta-zein in endoplasmic reticulum-derived protein bodies formed by beta-zein. AB - Zeins, the major seed storage proteins of maize, are of four distinct types: alpha, beta, delta, and gamma. They are synthesized on the rough endoplasmic reticulum (ER) in a sequential manner and deposited in ER-derived protein bodies. We investigated the potential for producing sulfur-rich beta-zein and delta-zein proteins in leaf and seed tissues by expressing the corresponding genes in a constitutive manner in transgenic tobacco. The delta-zein and beta-zein, when synthesized individually, were stable in the vegetative tissues and were deposited in unique, zein-specific ER-derived protein bodies. Coexpression of delta-zein and beta-zein genes, however, showed that delta-zein was colocalized in beta-zein-containing protein bodies and that the level of delta-zein was fivefold higher in delta-/beta-zein plants than in delta-zein plants. We conclude that delta-zein interacts with beta-zein and that the interaction has a stabilizing effect on delta-zein. PMID- 9338983 TI - Liquid chromatography with ultraviolet absorbance detection of ethylenethiourea in blood serum after microwave irradiation as an auxiliary cleanup step. AB - The report describes a method of deproteinizing serum that combines the action of acids with microwave irradiation. The acid concentration is ten times smaller than in the usual acid deproteinization method and the results are similar. The main advantages of the proposed method are the maintenance of the pH of the supernatant at around 5, and the reduction of the concentration of the deproteinizing agent. The procedure was applied to the determination of ethylenethiourea in serum samples by HPLC with spectrophotometric detection. Using 0.5 ml of serum, 0.04 microgram of ethylenethiourea were determined with recoveries between 89 and 109%. PMID- 9338984 TI - Infrared discrimination of enantiomerically enriched and racemic samples of methamphetamine salts. AB - A relatively rapid and simple means of enantiomer determination is described for the determination of methamphetamine, a common drug of abuse. The method employs the well known technique of infrared transmission spectrometry on solid samples dispersed within an alkali metal halide matrix. This approach exploits the solid state, ion-exchange reaction between methamphetamine hydrochloride and a potassium iodide matrix and the subsequent formation of the hydriodide salt in situ. The infrared properties of the hydriodide salt are distinct for enantiomerically enriched and racemic samples, and therefore are readily distinguished by infrared transmission spectrometry. This technique uses materials and instrumentation that are generally available to most crime laboratories. The applicability of this method to some other amine drugs is discussed. PMID- 9338985 TI - Application of near-infrared reflectance spectrometry in the study of atopy. Part 1. Investigation of skin spectra. AB - An investigation into the existence of spectral differences and differences in response in terms of water and lipid content between normal and atopic skin is described. Since NIR radiation penetrates complex structured matrices down to a depth of 0.15-0.20 mm, it is evident that the method lends itself to spectral detection of skin components down to the deepest level. First the reproducibility of readings made with the instrument was tested and it was also checked whether the use of the probe caused changes in skin equilibrium due to occlusion. Analysis of the NIR spectra did not enable normal and atopic subjects to be distinguished unequivocally but provided important information on the use of NIR spectrometry in these subjects and insights into the stratum corneum. Although the responses of water and lipid structures could not be read directly from the spectra, it was possible to decompose the global spectral information into components by principal components analysis. It was possible to observe a fraction of variance associated in different ways with water. PMID- 9338986 TI - Near-infrared reflectance spectrometry in the study of atopy. Part 2. Interactions between the skin and polyethylene glycol 400, isopropyl myristate and hydrogel. AB - An investigation into the existence of spectral differences and differences in response in terms of water and lipid content between normal and atopic skin after interaction with chemical agents is described. Three compounds were taken as models: a prevalently hydrophilic solvent (polyethylene glycol, PEG 400), a prevalently lipophilic solvent (isopropyl myristate) and a hydrophilic pharmaceutical (gel) used to promote contact in electrocardiography. Using principal component analysis it was possible to distinguish atopic and normal subjects by simple contact of the skin with chemical agents. PMID- 9338987 TI - Recovery of some common solvents from an adhesive commercial skin adsorption pad by thermal desorption-gas chromatography. AB - A thermal desorption-gas chromatography (GC) system was developed for use with commercial adhesive plasters used for monitoring exposure of hands to common solvents. The efficiency of solvent adsorption on the activated carbon pads located on the plasters was determined for acetone, trichloroethylene, D limonene, methanol, ethyl methyl ketone, toluene, tetrachloroethylene and m xylene. The degree of solvent recovery for the system was also investigated for each solvent, as was its sensitivity and reproducibility. All solvents exhibited > 90% adsorption on the pads at spiking levels of 100-200 ng for each solvent, except for m-xylene and d-limonene. Solvent recovery was dependent on the volatility of the solvent at spiking volumes of about 1 microliter per pad with solvents with boiling points above 110 degrees C showing recoveries of < 75%. Increasing primary desorption times and temperatures increased these values. The precision was good with RSD < 5% for all solvents over the range 0.5-5.0 microliters of applied solvent. It was possible to detect 15-60 ng of each solvent component within solvent mixtures on the pads with the exception of D limonene. It is concluded that all solvents tested except D-limonene can be determined on the pads under the conditions for thermal desorption-GC analysis described. The pads were used under protective gloves with six workers using xylene isomers as solvent in the workplace, when apparent solvent breakthrough through their gloves was observed. PMID- 9338988 TI - Miniaturized graphite sensors doped with metal-bathophenanthroline complexes for the selective potentiometric determination of uric acid in biological fluids. AB - Miniaturized poly(vinyl chloride) matrix membrane sensors in an all-solid-state graphite support, responsive to urate anion, were developed. The membranes incorporate lipophilic ion-pair complexes of urate anion with ruthenium(III), iron(II), nickel(II) and copper(I) bathophenanthroline (4,7-diphenyl-1,10 phenanthroline) counter cations. The sensors demonstrate a near-Nernstian response to urate over the concentration range 1 x 10(-2)-1 x 10(-5) mol l-1 and have micromolar detection limits and good selectivity properties. The response is virtually unaffected by pH changes in the range 7-10 and the response times are 5 10 s in aqueous solutions and in human serum and urine samples. A flow injection detector incorporating an iron(II) bathophenanthroline-urate graphite sensor was used for continuous monitoring of uric acid. The minimum detectable concentration was approximately 8 micrograms ml-1 and the sample throughput was approximately 120 h-1. Direct potentiometric determination of uric acid in the static and hydrodynamic modes of operation over the range 15 micrograms ml-1-1.5 mg ml-1 showed average recoveries of 98.7 and 97.8% with RSDs of 0.6 and 0.7%, respectively. Application of the method to the determination of uric acid in human serum and urine gave results that compared favourably with those obtained by the standard spectrophotometric method. PMID- 9338989 TI - Automated polarographic determination of sulfide as contaminant in parenteral amino acid solutions. AB - A method for the automated polarographic determination of sulfide in a gas diffusion flow system was developed. The volatile sulfide, existing as a contaminant in parenteral amino acid solutions, was measured after gas diffusion using 0.1 mol l-1 NaOH solution as acceptor. The linear range of calibration, for measurements at a dropping-mercury electrode (DME), was from 5 to 100 micrograms of sulfide with r = 0.999 and RSD = 7.5% (n = 5) for 10 micrograms of sulfide. For measurements at the hanging mercury-drop electrode (HMDE), with a preconcentration time of 30 s, the linear range of calibration was from 0.9 to 20 micrograms of sulfide with r = 0.998 and RSD = 5.8% (n = 5) for 2 micrograms of sulfide. Detection limits of 59 and 11 micrograms l-1 at the DME and HMDE, respectively, were calculated and the recoveries of sulfide from spiked samples were 91.5-104.5%. Parameters that affect the sulfide determination using this method, such as the organic content of the matrix, pH, flow rate and sample size, were investigated. PMID- 9338990 TI - Catalytic oxidation of uric acid at the polyglycine chemically modified electrode and its trace determination. AB - Cyclic voltammetry was undertaken to investigate the electrochemical behavior of uric acid at a polyglycine modified electrode. The modified electrode shows catalytic ability for the oxidation of uric acid, reducing the overpotential by 250 mV in pH 7.0 phosphate buffer solution. The enhanced voltammetric response can be used to determine uric acid. The linear range is between 5.0 x 10(-8) and 4.5 x 10(-6) M with a detection limit as low as 5.0 x 10(-9) M. The relative standard deviation is 1.4% (8 runs) at a concentration of 50 microM uric acid. The catalytic effect of the modified electrode resulted in the voltammetric resolution of the overlapping of uric acid and ascorbic acid. This allows the simultaneous detection of uric acid and ascorbic acid in the same sample. PMID- 9338991 TI - Cathodic stripping voltammetric determination of doxazosin in urine and pharmaceutical tablets using carbon paste electrodes. AB - Several voltammetric techniques were used to explore the reductive behaviour of the antihypertensive agent doxazosin on a bare carbon paste (CPE) and a Tenax modified carbon paste electrode (TMCPE). The results indicate that the process is irreversible and fundamentally controlled by adsorption, which allows doxazosin to be accumulated at the electrode surface. The cathodic adsorptive stripping (AdS) response was evaluated with respect to pH, accumulation variables and instrumental parameters, using differential-pulse (DPV) and square-wave voltammetry (SWV) as redissolution techniques. In both cases, a voltammetric peak close to 0 V in Britton-Robinson buffer (pH 6.6) was obtained after a preconcentration step at 0.55 V for 3 min (2000 rpm) and a subsequent cathodic scan. When the TMCPE was used, the limits of detection were 4.35 x 10(-11) and 5.18 x 10(-11) M for AdS-DPV and AdS-SWV, respectively. The deposition time (3 min) was improved relative to that obtained by means of CPE (6 min). Under the optimum operational conditions, the doxazosin reduction peak showed a linear response in the range from 6 x 10(-11) to 1 x 10(-9) M by using AdS-DPV, with an RSD of 3.39% for 3 x 10(-8) M doxazosin solution (n = 10). A method was developed for the determination of doxazosin in human urine and formulations. PMID- 9338992 TI - Enzymatic release of sequestered cows' milk fluoride for analysis by the hexamethyldisiloxane microdiffusion method. AB - The concentrations of diffusible and total fluoride in cows' milk samples from areas with widely different fluoride levels in drinking water were determined using a fluoride electrode. The diffusible fluoride was determined by direct hexamethyldisiloxane microdiffusion while for total fluoride, samples were subjected to either open ashing or digestion with proteolytic enzymes before microdiffusion. Magnesium nitrate was studied as a new fixative for milk during open ashing and compared with magnesium acetate. Diffusible fluoride ranged from 0.024 to 0.28 microgram ml-1 while total fluoride ranged from 0.05 to 0.31 microgram ml-1. The use of proteolytic enzymes before microdiffusion resulted in total fluoride measurement. It was concluded that all fluoride in milk is inorganic in nature with the bound fluoride being physically or chemically sequestered in the milk proteins. The proposed method is convenient for total fluoride analysis in milk. PMID- 9338993 TI - [Are nitrogen oxides major atmospheric pollutants?]. AB - Nitrogen oxides (N2O, NO and NO2) are at different levels important molecules in biological, ecological and public health terms. They are implicated in several fundamental planetary equilibria: climate and greenhouse effect, stratospheric and tropospheric ozone. NO is a very important cellular bio-mediator considered as weakly toxic for the pulmonary function, as N2O. The main toxicological questions are coming from NO2 whose toxicity has been studied with experimental (cellular, animal and human models) and epidemiological approaches. Its pulmonary toxicity appears relatively weak in animal and healthy human for concentrations in the order of 0.5-11 ppm or more. But NO2 nocivity must be considered for several reasons: first the inter-individual variability of the human response, second the increased sensitivity of asthmatics, chronic obstructive pulmonary population but also children and women, third the increased toxicity of NO2 associated with other (allergenic or not) aerocontaminants. These results lead to apply to NO2 a caution principle in indoor or outdoor air pollution; in the second case, WHO and European Union defined references values. PMID- 9338994 TI - [New anticancer agents derived from plants]. PMID- 9338995 TI - [Extemporaneous production of gas standard for the determination of carbon monoxide, hydrogen cyanide and ethylene oxide]. AB - This paper describes a new calibration method for headspace chromatographic assay of carbon monoxide, hydrogen cyanide and ethylene oxide. The calibration method is based on the in situ formation of gas in a closed system in order to avoid, the dangerous handling of pure gas or harmful substances. In the first method, the standardization is based on the in situ formation of carbon monoxide by the reaction of hot concentrated sulfuric acid with formic acid. In the second method, the standardisation is based upon the in situ reduction of ethyl thiocyano by dithiothreitol to produce HCN. In the third method, the standardisation is based upon the in situ chemical transformation of 2-chloro ethanol into ethylene oxide with a recovery of about 60%. PMID- 9338996 TI - [Cannabis: a current topic]. AB - The use of Cannabis, specially in Europe, is a very interesting subject for the media today. For several decades, Cannabis pharmacognosy has been more studied and new developments have appeared specially because of the cultivation of selected plants in the greenhouse and in Holland to obtain "nederwiet". Now, new research in the area of phytobiology, analysis and pharmacology has become necessary to bring scientific results for discussion on legalization of Cannabis consumption. The conclusion we can draw is that abuse of new forms by young people is dangerous for their health and disturb their behaviour. PMID- 9338997 TI - [Dermo-pharmaceutical efficacy of emulsions with NEO-PCL: galenic and rheological studies]. AB - The future EU prohibition (Directive 86/609/CEE) of using experimentation animals has induced the implementation of non-invasive techniques which are to substitute the present methods. Two non-invasive techniques are used to determine the cosmetological effectiveness of two emulsions formulated on a modern excipient NEO-PCL Base. Furthermore, the incorporation of multiflower honey is analyzed, being carried out the appropriate essays in order to obtain its tipification from a galenic and rheological point of view. After the formulation adjustment, O/W emulsioned systems, eudermic pH, and the rheological parameters to its application on the skin are obtained. Out of the effectiveness study, a high moisturization/emolliency degree is deduced. On the other hand, we conclude that multiflower honey rises the moisturizing power on this kind of products. PMID- 9338998 TI - [Non-invasive lesions of the cervical glands. Apropos of a study of 431 conization specimens]. AB - 431 cone biopsy specimens referred for CIN2 and CIN3 between 1984 and 1993 were reviewed with a peculiar attention paid to the possible associated endocervical glandular changes. The following features could be demonstrated: microglandular hyperplasia (17 cases), tubal metaplasia (15 cases), mesonephric hyperplasia (10 cases), in situ adenocarcinoma (7 cases), tunnel clusters (5 cases), and ectopic endometrium (4 cases). Cervical glandular atypia could not be found. A classification of the glandular lesions in uterine cervix has been proposed for an accurate diagnostic approach of lesions who could simulate carcinomatous changes. PMID- 9338999 TI - [Quantification of protein p53 expression by image analysis in 58 cases of colon carcinoma. Study of correlations between sex, age, histology, Dukes stage, DNA content and lymph node invasion]. AB - The analysis of p53 expression has been performed on 58 cases of colonic carcinomas. p53 expression was revealed by immunohistochemistry with the monoclonal antibody DO.7, and its quantification was performed by image analysis on deparaffinized tissue sections treated by microwaves. p53 expression evaluated by images analysis has been compared to visual estimation performed under light microscopy, and an excellent correlation was found between the two methods. No statistical significant relationships were found between the proportion of tumours expressing p53, sex, age (< 70 or > 70 years), histology (well, intermediate or poorly differentiated) and DNA index (< 1.3 or > 1.3) whereas the proportion of tumors expressing p53 was significantly higher in case of metastatic behaviour as well as in the C stage of Dukes classification. p53 expression was also significantly more important in colonic carcinomas with DNA index higher than 1.3 and in case of metastatic behaviour. According to these data, the metastatic behaviour is correlated both with the proportion of tumors expressing p53 and with the level of p53 expression. PMID- 9339000 TI - [Telepathology and "small cells" in cervical-vaginal smears: a new tool for diagnosis and teaching?]. AB - The identification of small cells in cervical/vaginal smears is usually considered as a difficult task, as well when they are examined through a classical light microscope or thanks to a teletransmission system. In this paper, we have compared both methods. 53 cases of smears with small cells were examined by two pathologists. The different types of small cells were described. The results and the discrepancies were studied. The reliability was rather good (45/53 cases, 84.6%). The main images were recorded onto a videotape for teaching purposes. PMID- 9339002 TI - [Paraganglioma of the cauda equina. Apropos of a case with review of the literature]. AB - Paragangliomas are unusual neuroendocrine tumors. The most common anatomical site is within head and neck (90%). It has rarely been described in the region of the cauda equina. We report the case of a 49 years-old white woman who presented with a slow, progressive cauda equina syndrome over a 20-years period. Radiological examination showed erosion of the vertebral laminae of L4, L5 and S1 and disclosed an intra-dural mass lesion occupying the entire spinal canal between L4 and S1. The patient underwent sub-total excision and adjuvant radiation therapy. Histological examination concluded to a paraganglioma. This diagnosis was confirmed by immunohistochemical studies. PMID- 9339001 TI - [Primary bone leiomyosarcoma. Anatomo-clinical case with immunohistochemistry study and review of the literature]. AB - We describe a primary leiomyosarcoma arising in the proximal part of the right tibia of a 38-year-old man. The diagnosis was confirmed by immunohistochemistry (positivity of tumor cells for alpha smooth muscle actin, HHF 35, desmin and vimentin). To the best of our knowledge, this is the 49th documented case of primary leiomyosarcoma of bone outside the facial skeleton. This exceptional tumor arises more commonly in adults (mean age: 53 years) and in the long bones of lower limbs, near the knee. The most frequent symptom is pain with or without swelling or fracture. Radiological findings invariably consist of a non specific osteolytic lesion. Although their histological appearance does not differ from that of extraosseous leiomyosarcomas, their diagnosis is difficult and often requires immunohistochemical and/or ultrastructural study. From a practical point of view, the diagnosis of primary leiomyosarcoma of bone also requires an intensive review of the case history and of previous pathology. This is necessary in order to eliminate an extraosseous primary site (mainly in uterus, gastrointestinal tract and soft tissues). As well as clinicopathological features, modes of treatment and results are also reviewed. PMID- 9339003 TI - Ovarian metastatic melanoma. A diagnostic pitfall in histopathologic examination. AB - Malignant melanoma metastasis to the ovary is exceptional. This observation relates the case of a 33 year-old woman who presented with a voluminous unilateral, histologically poorly differentiated, ovarian mass, and discusses the differential diagnosis. Immunohistochemical studies demonstrated strong positivity for S-100 protein, HMB-45 and vimentin. We emphasize the unpredictable clinical and biologic behavior of malignant melanomas and the necessity to perform immunohistochemical study by S-100 protein and HMB-45 in a poorly differentiated metastasis to the ovary, histologically compatible with malignant melanoma; even in the absence of intracytoplasmic pigment and clinical antecedent of cutaneous malignant melanoma. PMID- 9339004 TI - [Immature teratoma of the mandible with ameloblastic component]. AB - A case of immature teratoma involving mandible in a 56 year old woman is reported. This teratoma is composed of ameloblastoma structures mixed with some usual components of teratoma: osseous and cartilaginous foetal tissue, indifferenciated blastema, neuroblastic structures, acinous glands and melanotic cells. A lymph node is involved by metastatic mature glial tissue. This patient is treated by pelvimandibulectomy and irradiation. Histogenesis of this teratoma unusual by this ameloblastic component and this mandibular localization is reviewed. PMID- 9339005 TI - [Multifocal epithelioid angiosarcoma of the small intestine. Apropos of a case]. AB - A case of angiosarcoma in the small bowel is reported. There were multifocal lesions in the duodeno-jejunum with peritoneal metastasis, revealed by an hemorrhagic syndrome. This tumor rare in the gastro-intestinal tract was epithelioid. The authors present an histochemical and immunohistochemical study with a review of the literature. PMID- 9339006 TI - [Histopathology of protozoa]. AB - Protozoa are single-celled animals hosted by animals and humans. Numerous parasites can be considered to be non-pathogenic for humans but, in tropical countries, pathology caused by protozoa is a major public health problem. The study of parasitic protozoa in tissues has provided fertile ground for investigation. In practical diagnosis, this test is useful when the parasite is not eliminated in stools or absent in the blood. This review considers all the morphological criteria allowing an accurate identification of species. The new infectious agents, recently emerged with HIV infection are analysed by simple technical procedures for their identification. PMID- 9339007 TI - [Invasive papillary and pseudopapillary (micropapillary) carcinoma of breast]. AB - Classical invasive papillary carcinoma of the breast, is rare. In their pure form, they constitute 1.5% of all invasive breast carcinomas. Only one study clearly distinguishes them from intra-ductal papillary carcinomas; in this study they display a low metastatic potential and a relatively good prognosis. Lately, a new type of papillary carcinoma has been described which is characterized by inversion of the tumor cell polarity and papillations devoid of fibrovascular cores. Described by several authors by the term "micropapillary" carcinoma, the originally suggested term "pseudo-papillary" seems to be more appropriate; moreover, this designation would more clearly distinguish them from papillary carcinomas from which they clearly differ by their behavior. Furthermore, the pseudo-papillary carcinomas present an important lymphotropism, the effect of which on their clinical course is not yet clear. Further studies are necessary to determine whether these tumors, more frequent than classical papillary carcinomas, should be considered as a distinct clinical entity. PMID- 9339008 TI - [Flat adenoma. Endoscopic aspects]. AB - Flat adenomas are small, barely visible, raised lesions, sometimes with a central depression, which may be markers for a familial risk of cancer. Severe dysplasia is found in nearly half the cases. Carcinomas usually considered as occurring de novo may develop from flat adenomas. Detection of flat adenomas is difficult and substantially increases the duration of colonoscopy; therefore it should be reserved for high risk families. The stains and instruments needed to detect flat adenomas are detailed herein, as well as the excision technique. PMID- 9339009 TI - [Nosologic and anatomy pathological study of flat adenoma and associated lesions]. AB - Flat adenomas can be dysplastic from onset, whereas for polyps the risk of malignant transformation increases over time and with the size of the polyp. A dominant autosomal syndrome characterized by the development of flat adenomas has been reported. In a retrospective study of operative specimens from high-risk patients who had surgery for colorectal cancer, the author found that flat adenomas, which had been overlooked at initial evaluation, were a potentially useful phenotype. The mucosa adjacent to the flat adenomas showed hyperplasia and increased proliferation reminiscent of the morphologic abnormalities in aberrant crypts described by Pretlow; the latter have been found to occur rapidly in rats exposed to carcinogens and have also been demonstrated in patients at high risk for colon cancer. Aberrant crypt abnormalities and flat adenomas may be premalignant lesions. PMID- 9339010 TI - [Lymph node eosinophilic granuloma. Apropos of 2 cases of Langerhans-cell histiocytosis with isolated lymph node involvement]. AB - The authors report two cases of isolated lymph node involvement by Langerhans' cell histiocytosis which affected two young children. The histologic aspect reveals that lymph nodes have been modified by a proliferation of large histiocyte-like cells, associated with eosinophils. An immunohistochemical study on paraffin sections and for one case on frozen sections, reveals the usual phenotype of Langerhans' cells: these cells stain positively with S 100 protein and CD1 and are negative for both lysozyme and al antichymotrypsine. After a period of two years for one child and four years for the other, these children are in total remission, one spontaneously, the other after chemotherapy. PMID- 9339011 TI - TGF-beta 1 in colonic neoplasia. A genetic molecular and immunohistochemical study. AB - Transforming growth factor-beta proteins are key regulators of cellular growth and differentiation. To understand the role of TGF-beta in colonic tumour progression, 47 paraffin-embedded samples from colonic tumours (13 adenomas, and 34 adenocarcinomas) were studied. Gene mutations in the region coding for the active protein were studied by PCRSSCP analysis of exons 5, 6, and 7. TGF-beta 1 mRNA expression and localization were studied by NISH using cDNA probes generated by RT-PCR. Protein distribution was investigated by immunohistochemistry using antibodies against both intracellular and extracellular forms. Three mutations were found: one in exon 5 (Dukes C) and two in exon 7 (Dukes C and A). mRNA TGF beta 1 was observed in 9 (69%) of 13 adenomas and in 30 (88%) of 34 adenocarcinomas. Levels of TGF-beta 1 mRNA and proteins were higher in colorectal carcinomas than in colorectal adenomas. Tubular adenomas associated with dysplasia showed greater expression of TGF-beta 1 than adenomas without dysplasia and than non neoplastic colonic mucosa. In dysplasia and cancer, epithelial neoplastic cells and stromal cells were positive for TGF-beta 1. In normal tissue endothelial cells and granulocytes sporadically showed immunoreactivity for TGF beta 1, whereas epithelial cells were all negative. The three mutations in TGF beta 1 exons 5, 6 and 7 found in colorectal adenocarcinomas suggest that TGF-beta 1 mutation may play a role in colorectal carcinogenesis and that the presence of the mutant form contributes to the transformed state. Our two other findings, the loss of staining gradient in normal colonic mucosa and the higher levels of TGF beta 1 mRNA and proteins in colorectal carcinomas than in colorectal adenomas, indicate that the deregulation of TGF-beta 1 expression may occur as an early event in colorectal carcinogenesis. Although TGF-beta 1 expression is generally a reflection of cellular differentiation, tumour grade is not associated with mRNA TGF-beta 1 expression. As well as being present in the epithelial cells of the colonic tumours, mRNA TGF-beta 1 also occurred in the stroma. Its localization in the macrophages adjacent to tumour strongly suggests that TGF-beta 1 could be secreted by macrophages. This hypothesis should lead to new therapeutic strategies for colorectal carcinoma. PMID- 9339012 TI - [Cytologic aspects of cervical smears in optic microscopy in HIV seropositive women in Yaounde-Cameroon (Central Africa)]. AB - Cytologic aspects of cervical smears in HIV seropositive women in Yaounde Cameroon (Central Africa). The aim of this study was to present the cytologic aspects of cervical smears performed on HIV seropositive Cameroonian women and analysed by light microscope. Seropositive women (case group) and seronegative women (control group) had cervical smears which were stained by the Papanicolaou Method and analysed by light microscope. For the 65 seropositive women, there were: 62 inflammatory smears (95.5%), 2 normal (3%) and 1 low grade squamous intra-epithelial lesion (1.5%). The 50 seronegative women had: 35 inflammatory smears (70%), 13 normal (26%) and 2 low grade squamous intra-epithelial lesion (4%). In both groups, inflammatory smears were predominant. They were more frequent in seropositive women. There was no significant difference between the percentage of squamous intra-epithelial lesions in seropositive women (1.5%) and seronegative women (4%). We were unable to detect, in Cameroonian seropositive women, any specific lesions on cervical smears predictive of HIV infection without serology. PMID- 9339013 TI - [Granular cell tumor of the breast with lymph node neuro-naevic cell inclusions: a double pitfall to avoid]. AB - AIMS: To draw attention to the possibility of simultaneous occurrence of pitfalls in breast pathology. MATERIAL AND METHODS: Presentation of a curious case with an association of granular cell tumour of the breast and neuro-naevic inclusions in axillary lymph nodes, first interpreted as metastatic mammary carcinoma. DISCUSSION: Non malignant inclusions in axillary lymph nodes, rare but not unusual, may be erroneously interpreted as metastasis of carcinoma, establishing a wrong diagnosis of malignity in the breast. It is important to recognize this type of lesion and the possibility of their coexistence to correct the diagnosis. An approach to the histogenesis of this lesion is presented. PMID- 9339014 TI - [Metastasizing pleomorphic adenoma of the parotid. Apropos of a case]. AB - A case of metastasizing pleomorphic adenoma of the parotid in which both the primary tumor and metastasis were composed of benign pleomorphic structures is reported in a 55 year-old white woman. The pulmonary metastasis developed nine years after excision of the primary tumor without local recurrence. No mitotic activity or infiltrative growth pattern were present in the primary tumor. By flow cytometry, DNA content was diploid in both the parotid tumor and the pulmonary metastasis. Previously reported cases are reviewed. PMID- 9339015 TI - Malignant peripheral nerve sheath tumor with glandular differentiation. Report of a case with emphasis to the usefulness of immunohistochemistry in the differential diagnosis. AB - A case of malignant nerve sheath tumor with glandular differentiation in a patient with neurofibromatosis is presented. In the spindle celled areas the tumor cells reacted strongly with vimentin and S-100 protein, while the glandular epithelia reacted with EMA, cytokeratins (peptides 8, 19, and 20) and CEA, as well as a few of them also with chromogranin. Based on the results of immunohistochemical profile of the tumor cells, the differential diagnosis of this rare soft tissue tumor from biphasic synovial sarcoma with glands, as well as its distinction from any other spindle cell sarcoma with entrapped skin appendages could be greatly facilitated. PMID- 9339016 TI - [Developing cancer from a probable misdiagnosed pancreatic cystadenoma]. AB - This paper reports the malignant transformation of a benign pancreatic cyst treated by internal drainage in a 30-year-old woman. The initial lesion was diagnosed on surgical biopsy of the cyst wall. The diagnosis of pancreatic carcinoma was made after one year of satisfactory course, by fine needle cytologic aspiration of the fibrous retraction of the cyst; at that time the patient had severe pancreatic pain with multinodular liver on ultrasonography. The conclusion is the recommendation of total excision of pancreatic cysts whenever the diagnosis is incomplete or imprecise. PMID- 9339017 TI - [Subcutaneous Dirofilaria repens dirofilariasis]. AB - The authors describe a case of Dirofilaria repens subcutaneous dirofilariasis, in a resident of Corsica (France). Epidemiological and morphological features are related for various species of Dirofilariasis. These data, mainly based on clinical history, are usually sufficient to exclude other forms of subcutaneous filariosis, especially tropical. PMID- 9339018 TI - [A case of primary signet ring cell adenocarcinoma of the bladder]. AB - A case of primary signet ring cell carcinoma of the urinary bladder. This case of adenocarcinoma of the bladder including immunohistochemical study and autopsy demonstrates the diagnostic problems and the bad prognosis of these rare tumors. PMID- 9339019 TI - [Cutaneous semiology of febrile rashes in children]. AB - Generalized eruptions with fever are a frequent problem in paediatric practice. Clinical type of skin lesions, distribution and associated signs and symptoms are sometimes specific enough for a definitive diagnosis. However non specific clinical findings do not allow an aetiologic conclusion in some instances. More than 50 infectious agents, drugs and numerous inflammatory diseases are known to cause rashes in childhood. PMID- 9339020 TI - [Febrile eruptions and herpesviruses]. AB - There are 2 types of febrile skin eruptions due to herpesviruses: rashes (human herpesvirus 6 and 7. Epstein-Barr virus and cytomegalovirus), and vesicular eruptions (varicella and herpes zoster, primary and recurrent herpetic infections). Clinical diagnosis is often easy, based on the features of the eruption: characteristic sudden exanthem, clinical signs of infectious mononucleosis, or early vesicular trunk lesions characteristic of varicella or herpes. However, it must be stressed that exanthematic viroses can be highly variable. In some cases, and based on the background on which these eruptions occur, laboratory investigations will be necessary, according to the suspected virus. PMID- 9339021 TI - [Febrile purpura in children]. AB - The association of fever and purpura should first suggest the diagnosis of fulminant meningococcemia, owing to the severity of this condition. Compromise of peripheral circulation (cyanosis, prolonged refilling time) is important to consider, because normal blood pressure is usually observed at an early stage of a septic shock in children and particularly in young infants. When this hypothesis has been eliminated, other causes of febrile purpura can be considered: meningococcal meningitis; measles or other viral diseases; non infectious causes include mechanical purpura, Schonlein-Henoch's purpura and thrombocytopenia. In the frequent case where no cause has been found, the diagnosis of occult bacteriaemia should be considered, leading to parenteral administration of antibiotic following blood and cerebrospinal fluid cultures. PMID- 9339022 TI - [Measles and rubella]. AB - Measles is an acute disease characterized by fever, cough, conjunctivitis, erythematous maculopapular rash and pathognomonic enanthem. Vaccination had resulted in decrease of complications and mortality. But vaccination coverage in France is low, about 80%: the virus is always circulating and outbreaks in teenagers are possible. The recommendation of a booster dose at age eleven will contribute to reduce the incidence of the disease. Rubella is asymptomatic in 30 to 50% of infected children. There is a risk of transmission to pregnant women with negative serology. Reduction of virus circulation and immunization of young girls will result in decrease of the congenital rubella syndrome (65 cases/year in France). A vaccine booster dose at eleven in all children, combined with measles immunization, is necessary. PMID- 9339023 TI - [Cutaneous eruptions of streptococcal and staphylococcal origin]. AB - Scarlet fever consists in a diffuse exanthem associated with mucous changes. Classical scarlet fever is rare now, but other severe streptococcal infections have become more frequent, such as streptococcal toxic shock syndrome. The scarlatiniform exanthem and the shock observed in this disease are due to a streptococcal pyrogenic exotoxin. Exfoliative toxins secreted by Staphylococcus aureus are responsible for the tender erythema and cutaneous scaling characteristic of staphylococcal scalded skin syndrome of infancy. The so-called staphylococcal scarlet fever is probably an attenuated variant of this disease. Toxic shock syndrome toxin 1 (TSST1) is another staphylococcal toxin implied in the staphylococcal toxic shock syndrome. This disease is characterized by general symptoms and a scarlatiniform exanthem which are due to the effects of TSST1, acting as a superantigen. PMID- 9339024 TI - [Kawasaki syndrome]. AB - Kawasaki disease or mucocutaneous lymph-node syndrome has been described in Japan in 1967. Diagnostic criteria are clinical. The diagnosis must be evoked when a child less than 5-year-old has fever lasting for 5 days or more, particularly if associated with conjonctivitis, cheilitis, rash, or adenomegaly. Kawasaki disease is a vasculitis and can lead to lesions of the coronary arteries which are more frequent in younger infants. Intravenous gammaglobulins (in the first 10 days) lower the frequency of coronary lesions. Aetiology is still unknown but recent advances support the hypothesis that bacterial superantigens are involved. PMID- 9339025 TI - [Diagnosis of neonatal eruptions]. AB - The aetiological diagnosis of a skin rash in a neonate requires an accurate clinical analysis. Smear examination of blister fluid is sometimes necessary. Macular-papular eruptions include erythema toxicum and generalized infections, viral as well as bacterial. Pustular eruptions include benign transient dermatoses and infections. Bullous eruptions include staphylococcal toxic syndromes and hereditary epidermolysis bullosa. So, whatever the clinical appearance of the rash, the main problem is to document an infection and to implement early treatment. PMID- 9339026 TI - [Febrile toxiderma in children]. AB - Febrile cutaneous drug reactions in the child represent 6% of paediatric hospitalizations for dermatologic reasons. Diagnosis is difficult, for both infectious diseases and drug allergy can induce the same skin reaction. The same eruption can correspond to several drug-induced reactions. In a single child, there may be several causes of skin eruption and several drugs inducing similar cutaneous reactions. Clinical diagnosis and the method of clinical imputability lead to diagnosis. Paraclinical methods are of limited interest. Symptomatic treatment is begun on emergency admission. Upon identification, the responsible drug can be withheld and the authorities responsible for post-marketing surveillance can be notified. PMID- 9339027 TI - [Febrile eruptions in systemic diseases in children]. AB - During systemic diseases, various cutaneous lesions can arise. When the diagnosis of the disease is known, it is quite easy to attribute the cutaneous symptoms to this disease. But, when confronted to a child with a rash and fever infectious disease has to be excluded first. If the antiinfectious treatment is not effective, a systemic disease can be suspected. Acute systemic disease may present as malaise, fever and cutaneous lesions, without specific biological abnormalities. The cutaneous symptoms can then be precious in reaching the correct diagnosis. PMID- 9339028 TI - [Hemorrhage in the third pregnancy trimester. Diagnostic orientation]. PMID- 9339029 TI - [Hallucinations. Diagnostic orientation]. PMID- 9339030 TI - [Ankle sprains. Diagnosis, management in emergency situations]. PMID- 9339031 TI - [Dose-effect relationship in drug prescription]. PMID- 9339032 TI - [Exophthalmos. Diagnostic orientation]. PMID- 9339033 TI - [Transfusion of blood and blood derivatives. Measurements of safety, circulatory, immunologic and infectious risks and accidents]. PMID- 9339034 TI - The development of ophthalmoscopy. PMID- 9339035 TI - The current status of macular hole surgery. PMID- 9339036 TI - Measurements of foveal cysts in branch retinal vein occlusion. AB - This study was performed to quantify cystoid changes in the fovea using fluorescein angiography with a scanning laser ophthalmoscope and digital image analysis. In digitized angiograms of 50 patients with branch retinal vein occlusion the number of cysts, the area of each cyst, and the relation of the total area to visual acuity was determined. PMID- 9339037 TI - Indocyanine green angiography in central serous chorioretinopathy. AB - Five cases of various stages of central serous chorioretinopathy are presented. The indocyanine green angiographic characteristics of central serous chorioretinopathy are discussed. The value of this technique in recognizing central serous chorioretinopathy in older patients from genuine age related macular dystrophy will be indicated. PMID- 9339038 TI - Bromovinyldeoxyurdine treatment of outer retinal necrosis due to varicella-zoster virus: a case-report. AB - In December 1995, a 70-years old male was referred to us because of rapid visual loss in the right eye, one month after a central retinal artery occlusion in the left eye. This renal transplant patient, with limited renal function, was on immunosuppressive therapy. The diagnosis of bilateral progressive outer retinal necrosis (PORN) due to varicella-zoster virus (VZV) was confirmed by polymerase chain reaction (PCR) detection of VZV DNA in the aqueous fluid. As retinitis progressed despite of intravenous acyclovir administration, the antiviral therapy was switched to oral bromovinyldeoxyuridine (BVDU). This case-report demonstrates that oral BVDU can be a good alternative to acyclovir for the treatment of VZV retinal infections. PMID- 9339039 TI - Ophthalmological signs of tuberous sclerosis. AB - A patient with tuberous sclerosis and retinal astrocytic hamartomas is described. The different types of these astrocytomas are illustrated as well as systemic signs of Bourneville disease. Finally the value of the ophthalmological examination in the diagnostic work-up prior to genetic counselling the patient and/or the family members is discussed. PMID- 9339040 TI - Vectoranalysis of surgically induced astigmatism in small corneal and scleral cataract incisions. AB - Vector analysis of surgically induced astigmatism was performed on several scleral and corneal tunnel incisions (6.0-mm W; 3.5- and 2.5-mm L scleral tunnel; 4.1-mm linear and 2.5-mm L corneal tunnel). The analysis revealed a mean astigmatism between 1.18 and 0.64 D, as compared to 0.35 D in a control group of nonoperated eyes. The best results were obtained with the scleral and corneal L shaped incisions. PMID- 9339041 TI - A proper role for organized medicine: the AMA response. PMID- 9339042 TI - Do today's medical residents really have it better? PMID- 9339043 TI - Can the patient make treatment decisions? Evaluating decisional capacity. PMID- 9339044 TI - Assessing and minimizing reproductive risks of cancer chemotherapy. PMID- 9339045 TI - Diagnosing and treating hallux valgus: a conservative approach for a common problem. AB - For most patients with hallux valgus, the problem is caused by wearing shoes that are too tight, and conservative measures can help. We review how primary care physicians can evaluate and treat this problem, and when to refer to an orthopaedic surgeon. PMID- 9339046 TI - Ethical perspectives on Jehovah's Witnesses' refusal of blood. AB - When Jehovah's Witnesses refuse blood, they regularly ask their physicians to explore and provide all other medically and scientifically based alternatives, even when these alternatives may not be as effective and may carry risk of failure that could lead to physical disabilities or death. An awareness of the values at stake and of other cases from personal experience and the literature can help physicians, patients, and their families to move toward ethically responsible decisions and actions. PMID- 9339047 TI - Travel medicine for the primary care physician. AB - Each year millions of people travel overseas, where they may come into contact with infectious diseases unfamiliar to citizens of the industrialized world. Reviewing the potential risks, and how to avoid them, greatly enhances the chances of an uneventful trip. This article contains specific recommendations including recipes for oral rehydration solutions that travelers can prepare. PMID- 9339048 TI - New drugs for reducing cardiovascular risk in women. AB - Atherosclerosis is largely preventable in women. Clinicians need to appreciate the gender-associated risks of cardiovascular disease and emphasize to their women patients that life-style changes can reduce cardiovascular risk. However, newer oral agents for diabetes and the statins for hyperlipidemia are important pharmacological adjuncts. PMID- 9339049 TI - History of ophthalmology in Hawaii. PMID- 9339050 TI - Glaucoma. PMID- 9339051 TI - Flashes, floaters retinal tears and retinal detachment. PMID- 9339052 TI - Retinopathy of prematurity. PMID- 9339053 TI - Diabetic retinopathy. PMID- 9339054 TI - Loss of reading and central vision due to macular diseases--therapeutic management, advances and limitations. PMID- 9339055 TI - Advances in ophthalmic plastic, reconstructive and orbital surgery in Hawaii. AB - Advances in ophthalmic plastic and reconstructive surgery continue to be made in the state of Hawaii. A new technique of dacryocystorhinostomy using the endoscope and the pulsed Holmium: YAG laser as well as a subconjunctival endoscopic assisted approach to orbital surgery for orbital decompression is described. The integrated hydroxyapatite implant replacing the enucleated eye gives excellent postoperative extraocular motility. PMID- 9339057 TI - Ophthalmology in Hawaii 1997 and beyond. PMID- 9339056 TI - Corneal & refractive surgery. AB - Excimer lasers and high technology instrumentation have ushered in a new era of vision improvement surgery in Hawaii, replacing the more traditional forms of refractive surgery: cataract surgery, corneal transplant surgery, and radial keratotomy. Corneal surgery has been enhanced by new techniques of microsurgery and a more effective tissue procurement system for donor corneal tissue. Several laser centers provide the latest in FDA-approved excimer laser procedures including PRK and PTK. Mild to moderate myopia and astigmatism may now be corrected. Off-label use of LASIK, too, may soon be realized. PMID- 9339058 TI - Medicine in antebellum Mississippi. PMID- 9339059 TI - An approach to reducing infant mortality rate through the utilization of lay home visitors. PMID- 9339060 TI - Atrial fibrillation quality improvement. PMID- 9339061 TI - Correlates of the quality of life of adults with severe or profound mental retardation. AB - To ensure quality of services for individuals with mental retardation/developmental disability, professionals must measure consumer outcomes related to lifestyle. In this study variables contributing to quality of life for 60 adults with severe or profound disabilities who resided in ICF/MR community-based homes for 4 to 5 persons were examined. Using the Quality of Life Index, we studied interrelations among personal lifestyle characteristics of adults and community-home program characteristics with quality of life factors. The R2 effect size (.571) involving total scores on the Quality of Life Index as the criterion variable was large and statistically significant. PMID- 9339062 TI - Adults with mental retardation as supports to their parents: effects on parental caregiving appraisal. AB - This study was conducted to determine whether support provided to caregivers by their adult children with mental retardation would influence caregiving appraisals. We also examined how severity of disability of the adult child, personal and social resources of the caregiver, and amount of caregiver assistance to the adult with mental retardation influenced caregiving appraisals. Using surveys and interviews we collected information from 80 primary caregivers on caregiving burden and satisfaction and six predictors of burden and satisfaction. Findings indicate that greater support from the adult child to the caregiver resulted in greater satisfaction and less burden. Adaptive and maladaptive behaviors and caregiving assistance all predicted caregiving satisfaction but only maladaptive behaviors predicted caregiving burden. PMID- 9339063 TI - "I want to talk like everyone": on the use of multiple means of communication. AB - This qualitative case study is a description of a young man with autism who communicated using speech, sign language, facilitated communication, body language, and his mother's conversational supports. Participant observation, interviews, and review of records were used to explore his current and past communication practices. These practices illustrate his preference for speaking and the complexity of choosing among communication means on an ongoing basis. Although the young man and his mother differed in their thinking about communication, they revealed a common goal: for Michael to participate in ordinary life activities as a member with a voice. PMID- 9339064 TI - Congruence and predictive power of mothers' and teachers' ratings of mastery motivation in children with mental retardation. AB - The congruence between mothers' and teachers' ratings of mastery motivation among 3-year-old children with mental retardation was investigated. The extent to which maternal and teacher ratings of task persistence at entry to preschool are predictive of observed mastery behaviors at age 5 was tested. Results indicate that mothers rated their children's task persistence behaviors higher than did teachers. Further, once the child's cognitive level and teacher ratings were controlled for statistically, maternal ratings of the child's mastery behaviors were predictive of the child's task mastery performance 2 years subsequent. Implications for educational planning were discussed. PMID- 9339065 TI - Support, problem-solving/coping ability, and personal burden of younger and older caregivers of adults with mental retardation. AB - Relations between service and support utilization, problem-solving/coping strategies, level of personal burden experienced by younger and older caregivers were examined. Overall, there were no differences in the number of support services received. However, younger caregivers reported significantly more unmet service needs and rated significantly more of them as a critical or an emergency need. Both groups had highly developed effective problem-solving skills. However, older caregivers were more likely to seek spiritual support and the younger caregivers more apt to mobilize their families to acquire and accept help. Older caregivers experienced significantly less personal burden. Results suggest that younger caregivers are more predisposed toward seeking outside help and have higher expectations of the service system. PMID- 9339066 TI - Presidential address 1997--benchmarks for the next millennium. PMID- 9339067 TI - Science, disability, and voice: a response to Danforth. PMID- 9339068 TI - A rift not that severe: science and hope: a response to Danforth. PMID- 9339069 TI - Laura Bridgman, mental retardation, and the question of differential advocacy. PMID- 9339070 TI - The PSDA and conflicts with foreign MTFs. PMID- 9339071 TI - Contaminated dispensers increase risk of infection. PMID- 9339072 TI - Reservist fitness evaluations. PMID- 9339073 TI - The Association of Military Surgeons of the United States Constitution and Bylaws. Association of Military Surgeons of the United States. PMID- 9339074 TI - Health effects of the stressors of extreme environments on military women. AB - This article examines the health effects of generic and unique stressors on military women's health. It is an outgrowth of work performed under the Defense Women's Health Research Program and participation in the Forum on the Health of Women in the Military held at the Uniformed Services University of the Health Sciences on June 17 to 19, 1996. We review gender differences in the effects of stress on health. We comment on some of the methodological challenges in researching gender effects. We hypothesize about some of the ways in which military women may be more vulnerable to specific stressors and/or use different coping strategies than their male counterparts. Finally, we make recommendations about military training and future research. PMID- 9339075 TI - Patterns and risk factors for exercise-related injuries in women: a military perspective. PMID- 9339076 TI - Women in the Persian Gulf War: health care implications for active duty troops and veterans. AB - The health of women who participated in Operations Desert Shield/Storm was evaluated to better understand the medical requirements of deployed military women and women veterans of the Persian Gulf War. Women's health care needs during the Persian Gulf War were reported to be very similar to those of men, with the exception of gynecologic problems, which generally were not serious and did not require hospitalization. However, insufficient data were obtained to identify specific health care needs among deployed women troops. During the 5 years since the end of the Persian Gulf War, no unique health problems have been identified among women veterans. Whether there will be any exceptional long-term health care requirements currently is unknown. Nevertheless, important medical problems of all women-reproductive issues, menopause, osteoporosis, joint disease, breast cancer, heart disease, and stroke-inevitably will be major considerations when caring for this population of war veterans. PMID- 9339077 TI - Post-traumatic stress disorder and functioning and quality of life outcomes in female Vietnam veterans. AB - OBJECTIVE: This investigation assessed whether current post-traumatic stress disorder (PTSD) was associated with impaired functioning in a nationally representative sample of female Vietnam veterans. METHODS: Logistic models were used to determine the association between PTSD and outcome while adjusting for demographic characteristics and medical and psychiatric co-morbidities. RESULTS: PTSD was associated with significantly elevated odds of poorer functioning in five of the six outcome domains; only the association between perpetration of violence in the past year and PTSD did not achieve statistical significance. After adjusting for demographics and medical and psychiatric co-morbidities, PTSD remained associated with significantly elevated odds of bed days, poorer physical health, and currently not working. CONCLUSIONS: Among female Vietnam veterans PTSD is associated with a broad profile of functional impairment. The significantly increased odds of impaired functioning and diminished quality of life suggest that PTSD may be the core problem of the set of problems afflicting female Vietnam veterans. PMID- 9339078 TI - Shipboard women's health care: provider perceptions. AB - Women have served aboard auxiliary U.S. Navy ships as integrated members of the shipboard work force since 1978. In 1994, women first started serving aboard combatant ships with the inclusion of women in the work force of the aircraft carrier USS Dwight D Eisenhower (CVN 69). The provision of the highest standard of medical care for both men and women is a priority at all levels in the U.S. Navy. This study assesses the perceptions of shipboard health care providers regarding their ability to provide adequate women's health care. This evaluation was performed by conducting a personal interview with the senior health care provider of each of 32 ships on which women are integrated members of the work force. Medical department representatives reported that most ships have training programs for birth control (90.6%), sexually transmitted diseases (96.9%), and the Navy pregnancy policy (84.4%). Health care providers also reported limitations in available supplies (i.e., contraceptives, pregnancy tests, and sexually transmitted disease tests). PMID- 9339079 TI - Changes in women's health care aboard one ship. AB - Health care needs of women assigned to sea duty may change over time. OBJECTIVE: Determine changes in the obstetrical/gynecological needs of U.S. Navy women assigned to a submarine tender. METHODS: Retrospective record review of personnel aboard one U.S. Navy ship in 1990 and in 1995. RESULTS: The demographic character of the female crew members changed. Compared with 1990, the women in 1995 were older, more experienced, of higher rank, more likely to use contraception, and more likely to have children. The pregnancy rate dropped from 2.7 to 1.5 per 100 women per month. Sexually transmitted diseases were less frequent, and the "satisfactory Papanicolaou smear" rate increased from 52 to 93%. Both in 1990 and 1995, women utilized Sick Call more often than men. CONCLUSION: Measurable changes occurred in the obstetrical/gynecological health care needs of women assigned to one ship in 1990 and 1995. PMID- 9339080 TI - Field-expedient gynecologic examination table. AB - In field operational environments, the gynecologic health needs of women may be difficult to provide because of the lack of a small, lightweight, durable, inexpensive gynecologic examination table. Such a table already exists, in pieces, in the inventory of most field-deploying units of battalion aid station size or larger. Because the table's existence is not commonly known, we describe the assembly and use of this field-expedient gynecologic examination table. PMID- 9339081 TI - Genital mutilation of young girls traditionally practiced in militarily significant regions of the world. AB - Very few physicians practicing in the United States have experience in treating female patients who have undergone mutilation of the external genitalia, incorrectly termed female circumcision. This procedure, known as infibulation, consists of removing the clitoris, prepuce, and portions of the labia of young girls, usually younger than 7 years of age. Infibulation has been practiced by lay midwives for centuries in the Horn of Africa and in other African and Middle Eastern countries. This paper discusses infibulation, the techniques, and the recommended medical and obstetric management of patients subjected to genital mutilation. With increased immigration to the United States by Africans and Middle Easterners, and with readily increasing military medical deployments, primary care physicians and specialists can expect to be confronted with patients who have undergone this disfiguring procedure during their youth. PMID- 9339082 TI - Fine needle aspiration of palpable breast masses performed in a military obstetrics/gynecology clinic: a follow-up report. AB - Evaluation of breast disease has increasingly become more integrated into the routine gynecology care of women seen in the obstetrics/gynecology (OB/GYN) clinic. Patients expect their obstetrician-gynecologist to have expertise in evaluation and diagnosis of breast problems that arise from self examination, routine mammography, unusual breast symptoms, or clinical findings during annual gynecology examinations. In 1993, Tripler Army Medical Center Department of Obstetrics and Gynecology initiated a Breast Evaluation Clinic to better serve its patients with breast problems and to train military OB/GYN resident physicians in evaluation and diagnosis of breast disease. A preliminary report of the first 40 patients evaluated in this Breast Evaluation Clinic was previously published in Military Medicine. The patient evaluation, the technique of performing fine needle aspiration (FNA) of breast masses, and the cytologic slide preparation was described in the preliminary report. This follow-up report presents a total of 245 patients who underwent FNA of palpable breast masses in the Tripler Army Medical Center OB/GYN Department Breast Evaluation Clinic between December 1, 1993, and December 8, 1995. Patients found to have suspicious breast masses or abnormal mammography reports at the time of evaluation were immediately referred to the Department of General Surgery for evaluation rather than be subjected to FNA in the OB/GYN Department Breast Evaluation Clinic. Of the 245 patients who underwent FNA, 26 (11%) were referred to the Department of General Surgery for treatment or open biopsy based on cytologic diagnosis and evaluation in the OB/GYN Breast Evaluation Clinic. No major complications from the FNA procedures occurred during this 2-year study period. PMID- 9339083 TI - The practice of obstetrics in Mongolia. PMID- 9339084 TI - Sexually transmitted diseases: risk behaviors of female active duty U.S. Army recruits. AB - This study examined the sexually related behaviors of 594 unmarried female Army recruits at the beginning of their basic training. The response rate for the entire sample of 818 recruits was 99.4%, with data from the 224 married recruits being eliminated from the analysis. The recruits completed an anonymous survey examining several independent variables: frequency of condom use, number of sexual partners, screening behaviors, and pregnancy and sexually transmitted disease prevention methods. The findings are discussed with regard to suggested appropriate and cost-effective educational interventions. PMID- 9339085 TI - Behavioral treatment of exercise-induced urinary incontinence among female soldiers. AB - One-third of 450 female soldiers surveyed indicated that they experienced problematic urinary incontinence during exercise and field training activities. The other crucial finding of this survey was probably that 13.3% of the respondents restricted fluids significantly while participating in field exercises. Although only 5.3% of respondents felt that their urine leakage had a significant impact on their regular duties, it is obvious that many more are sufficiently worried about leakage to put themselves at significant risk for dehydration-related injuries. This study tested whether behavioral interventions effective among older people could help younger soldiers. Thirty-nine female soldiers reporting exercise-induced urinary incontinence underwent urodynamic assessments of bladder capacity, urethral closure pressure, and detrusor contraction pressures as well as a symptom questionnaire before and after therapy. They were stratified by diagnosis of physical stress incontinence or mixed urge/stress incontinence and randomized into two groups. Twenty-three participants performed pelvic muscle exercises with urethral biofeedback for 8 weeks, and 16 participants performed pelvic muscle exercises alone. Patient reports as well as the post-treatment examinations indicated that all subjects improved significantly. Only five subjects in the biofeedback/exercise and three in the exercise-only group desired further treatment. All subjects initially diagnosed with detrusor dysfunction had normal readings at the end of the study. Thus, behavioral treatments can be effective against exercise-induced urinary incontinence among most female soldiers. PMID- 9339086 TI - Female United States Air Force pilot personality: the new right stuff. AB - With increasing numbers of female military pilots, it is important to understand the psychological and psychiatric gender differences of pilots. Using the "big five" personality structure (neuroticism, extraversion, openness to new experiences, agreeableness, and conscientiousness), female United States Air Force pilots were compared with both male Air Force pilots and to a female comparison group. Female Air Force pilots were higher on the Extraversion, Agreeableness, and Conscientiousness scales than male pilots. Female pilots were also higher on these scales than the female comparison group and lower on the Neuroticism and Openness scales than that comparison group. It is suggested that these traits are highly adaptive for Air Force pilots, given the nature of modern military operational requirements. PMID- 9339087 TI - Risk factors associated with stress reactions in female Marines. AB - Women have a higher stress fracture rate than men in military studies, although the exact cause of this is not clear. Hyperpronation has been implicated as a potential risk factor for injury. In this prospective observational study, we measured subtalar joint range of motion in 101 women (ages 20-27 years) enrolled in Marine Corps Officer Candidate School in June 1994. The purpose of this study was to identify risk factors for injury in female Marine Corps officer candidates. The primary area of interest was the association between the amount of subtalar joint range of motion and stress reactions. Questionnaires were administered that explored previous physical activities, sports participation, and menstrual history. Anthropometric measurements were performed, including subtalar joint range of motion. During the 10 weeks of physical training, 11.5% of the women (N = 12) had stress reactions compared with 7% of the men (N = 10). There was no statistically significant difference in the means of subtalar joint range of motion in the stress reaction group compared with the non-stress reaction group. Differences in stress reaction rate across quartiles of subtalar joint range of motion were not significant. Those women who ran fewer miles (< or = 2.8 miles per session) before training had a higher rate of stress reactions (p < 0.04). Younger individuals (< 23 years) had a higher rate of stress reactions (p < 0.01). Women with fewer menstrual periods (< 10 per year) had a higher rate of stress reactions (p < 0.02). A narrow pelvis (< or = 26 cm) was associated with a higher rate of stress reactions (p < 0.09). We conclude that an increased subtalar joint range of motion is not a risk factor for stress reactions in women. However, further studies with a larger study population should be performed to confirm these findings. PMID- 9339088 TI - Prevalence and contributing factors of eating disorder behaviors in a population of female Navy nurses. AB - Eating disorders continue to be reported at a steady rate among lay women. The disorders of anorexia nervosa (AN), bulimia nervosa (BN), and eating disorders not otherwise specified (NOS) have only been reported in a military population in isolated case reports. The military lifestyle and the nursing profession mimic the environment that appears to prevail among women with eating disorders who have been previously studied in civilian populations. A total of 706 active duty female Navy nurses returned an anonymous mailed survey that was developed to correlate military and professional demographics and variables with current and past eating disorder behaviors. Responses were analyzed using the criteria established by the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders Third Edition, Revised; for AN, BN, and NOS eating disorders. Nurses who fit the criteria for AN were 1.1%, and 12.5% met the criteria for BN. NOS was seen in 36% of those sampled, and those classified as normal by exclusion were 50.4%. The Navy provides an environment in which eating disorder behaviors thrive and survive due to many reported military and professional factors. PMID- 9339089 TI - Factors influencing burnout and job stress among military nurses. AB - Burnout among military nurses has been found to lead to job absenteeism, staff conflicts, and a high turnover of personnel. Factors influencing nurses working in smaller and often isolated military installations of the South African National Defence Force were investigated using a job-stress and burnout questionnaire and a semi-structured interview. Investigation focused on registration categories, geographic location, and age. It was found that the senior registration categories experienced more burnout, and nurses in isolated areas reported almost double the number of cases of burnout than nurses in larger centers. Age played a role in the very young (19-25 years) and older (40-50 years) nurses. The lack of support from supervisors, high responsibility, long working hours, and task overload were the four most common stressors reported. Some suggestions are forwarded to manage the risk of burnout among military nurses in similar situations. PMID- 9339090 TI - Reevaluation of blood transfusion strategies. PMID- 9339091 TI - Evidence-based medicine. PMID- 9339092 TI - Painful oral ulcers with hydroxyurea therapy. AB - Five patients treated with hydroxyurea for various haematological malignancies developed multiple painful oral ulcers. Their neutrophil counts were either normal or elevated. The ulcers disappeared with cessation of hydroxyurea. Oral ulcers recurred when hydroxyurea was resumed in one of the patients. As the patients were unable to tolerate this painful side effect, hydroxyurea had to be discontinued. Appearance of painful oral ulceration seems to be independent of dosing rate or total cumulative dose of hydroxyurea. PMID- 9339093 TI - Penile revascularisation for vascular impotence. AB - OBJECTIVE: Many methods of microscopic penile revascularisation procedures have been employed over the past 2 decades for the treatment of vasculogenic impotence, with varying success rates. The aim of our study was to evaluate the effectiveness and complications of deep dorsal vein arterialisation in the treatment of selected patients with arteriogenic and mixed arteriogenic/venogenic impotence. METHODS: This involved a retrospective analysis of 6 patients with vasculogenic impotence who presented to Toa Payoh Hospital from December 1991 to November 1994 and had penile revascularisation surgery performed. All patients underwent an extensive preoperative assessment, including dynamic infusion cavernosometry and cavernosography and selective pudendal arteriography. RESULTS: The 6 patients were aged between 27 and 51 years (mean 44 years). 2 (33%) patients had pure arteriogenic impotence, while 4 (66%) had mixed arteriogenic and venogenic impotence. Two patients (33%) had excellent surgical outcomes and 2 patients (33%) were considered improved. The mean follow-up period was 19.8 months (range 8 to 37). Complications were minimal. CONCLUSIONS: We conclude that although the results of penile revascularisation are promising in carefully selected patients, further studies with longer follow-up and more objective postoperative tests of hemodynamic and erectile function are needed to assess the true value of this mode of treatment. PMID- 9339094 TI - A series of ovarian clear cell and endometrioid carcinoma and their association with endometriosis. AB - AIM: To present our department's experience with clear cell carcinoma and endometrioid carcinoma of the ovary, paying particular attention to their relationship with endometriosis and concomitant endometrial pathology. METHOD: Retrospective review of case records. RESULTS: From July 1986 to March 1995, 11 patients with clear cell carcinoma and 20 patients with endometrioid carcinoma of the ovary were treated. Of the patients with clear cell carcinoma, five (45%) had associated endometriosis. One patient (9%) also had endometrial adenomatous hyperplasia. Of the 20 cases of ovarian endometrioid carcinoma, four (20%) had endometriosis present during histopathological examination. Six patients (30%) had concomitant endometrial pathology (five cases of endometrial carcinoma and one with adenomatous hyperplasia). CONCLUSION: Our series shows that the clear cell ovarian carcinoma may often be associated with endometriosis, more so than the endometrioid type of ovarian carcinoma. However, the patient with ovarian endometrioid carcinoma may also harbour a concurrent endometrial pathology. PMID- 9339095 TI - A randomised double-blind study of vaginal misoprostol vs dinoprostone for cervical ripening and labour induction in prolonged pregnancy. AB - BACKGROUND: Dinoprostone, is presently used in our standard protocol for cervical ripening and labour induction. In search for a cheaper alternative, misoprostol has been found to be a good substitute. In view of the potential saving it might offer, we set out to test its efficacy against the standard dinoprostone. METHODS: A randomised double-blind study involving 50 pregnant women with prolonged pregnancy, treated at a government hospital in Malaysia, was carried out. Two hundred micrograms of intravaginal misoprostol were compared with 3 mg of dinoprostone in each treatment arm. RESULTS: In the misoprostol group, labour was successfully established in 92% of cases compared to 64% in the dinoprostone group (p = 0.04). The induction-delivery interval was shorter with more women delivering within 12 hours (72% vs 28%, p = 0.047). Maternal and neonatal complications, mode of delivery, the need for oxytocin and pethidine were quite similar statistically. Polysystole was more frequent (28% vs 12%, p = 0.28) in the misoprostol group but it was not associated with fetal distress. CONCLUSION: The study showed that misoprostol was a more effective drug in labour induction. PMID- 9339097 TI - Neurocysticercosis--case report and literature review. AB - A 30-year-old Indian migrant worker presented with seizures at the National University Hospital. A CT-scan of the brain showed multiple calcifications and cysts consistent with neurocysticercosis. Plain radiographs of the humeri and femora also revealed multiple soft tissue calcifications. He was given a course of anti-helminthic therapy and started on anti-epileptics. Neurocysticercosis is a common cause of seizures in endemic areas and must be considered in the differential diagnosis of epilepsy involving the members of the large community of migrant workers in Singapore. PMID- 9339096 TI - Reconstruction of a medial tibial plateau defect using a "pillar" bone graft--a report of two knee reconstructions. AB - A technique of reconstructing a large tibial plateau defect has been described using a solid bone graft as a pillar to hold up the tibial component of a total knee replacement. The advantage of using a solid bone graft is that there is good initial structural support for the tibial component. This enables early mobilisation. This method has worked well in the two knees reported. Other methods of reconstructing a tibial plateau defect are discussed. PMID- 9339098 TI - Suxamethonium and cardiac arrest. AB - We report a case of cardiac arrest due to hyperkalaemia following administration of suxamethonium during a procedure to facilitate a change of endotracheal tube in a septic patient. The cause of this rare but fatal complication is briefly described and discussed. In view of this, suxamethonium should be used with great caution in patients with burns and other forms of physical injury, in a number of nervous system disorders, and in critically ill patients requiring prolonged ITU care. PMID- 9339099 TI - Clinics in diagnostic imaging (27). Sarcoidosis. AB - A 44-year-old Caucasian man presented with third-degree heart block. Chest radiograph and high-resolution computed tomography (HRCT) of the thorax showed mediastinal and bilateral hilar lymphadenopathy associated with a diffuse, bilateral micronodular pattern. The HRCT findings and differential diagnosis of sarcoidosis are reviewed. PMID- 9339100 TI - What you need to know: spousal abuse (I). PMID- 9339101 TI - What you need to know: spousal abuse (II). PMID- 9339102 TI - A brief introduction to the early history of surgery in Singapore (Part II). PMID- 9339103 TI - The manpower question. PMID- 9339104 TI - Consent for and confidentiality of HIV testing. PMID- 9339105 TI - Laparoscopic antireflux surgery--technique and results. AB - Although gastroesophageal reflux disease (GERD) can be effectively treated by proton-pump inhibitors, surgery is still the only means of definitive cure of the disease. After introduction of laparoscopic surgery, there has been a clear trend to surgical repair of the incompetent cardia. The indications for surgical treatment are: endoscopically proven esophagitis, persistent or recurrent complaints under medical treatment, esophageal stricture and/or pH-metrically proven acid reflux as well as reflux-induced coughing (chronic aspiration). Although the laparoscopic antireflux operations is a technically demanding procedure, it can be performed with similar results as compared to conventional surgery. The operative technique is reported in detail. From January 1992 to March 1997, 146 consecutive patients with GERD have been operated on laparoscopically. The overall conversion rate was 8.2% (n = 12). 133 patients were operated on according to the Nissen procedure including hiatoplasty. The Toupet operation was performed in only one case. 84 men and 42 women had a mean age of 49 years (20-76). The median duration of symptoms was 48 months (1-600). Except five patients all had medical treatment for at least 2 years. Twice pneumatic balloon dilatation of an esophageal stricture was necessary preoperatively. The median operation time was 210 minutes (70-660). Conversion to open surgery because of intraoperative complications was necessary in 6 patients. Postoperative complications occurred in 14 patients, all of them being successfully treated conservatively. No patient died. 121 patients (90.3%) had follow up examinations for at least 6 months. Retreatment was necessary in 5 cases: 1x slipped Nissen (laparoscopic repair), 1x intrathoracic hernia (conventional reoperation), 2x dysphagia > 4 months postoperatively (endoscopic balloon dilatation) and 1x recurrent ulcer (conventional operation). With a correct indication, laparoscopic Nissen repair for GERD is a suitable, safe and definitive treatment. PMID- 9339106 TI - Histiocytic syndromes in children. AB - The "histiocytes" are a group of proliferative disorders of the mononuclear phagocyte system whose etiologies are basically unknown. The majority of childhood histiocytoses are expressions of excessive numbers of Langerhans cells, representing so-called Langerhans cell histiocytosis. Fifteen patients who were diagnosed with histiocytosis syndrome at the Pediatric Hematology and Oncology Department of Ege University Hospital between October 1986 and January 1995 were included in this study. The majority of the patients had Langerhans cell histiocytosis (LCH), and skeletal involvement was the most common manifestation. A good response to radiotherapy and chemotherapy was obtained by our patients with unifocal and multifocal involvement of LCH. Two patients with disseminated LCH died with progressive disease. In the patient with Rosal-Dorfman disease, a partial response was obtained with prednisone. The patient with malignant histiocytosis died during a relapse at the end of one year. Organ dysfunction and the patient's age are important factors affecting the outcome of the disease. PMID- 9339107 TI - Granulocyte and granulocyte-macrophage colony-stimulating factors, their receptors and interleukin-3 levels in newborns. AB - Serum levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and interleukin-3 (IL-3) as well as neutrophil counts were studied on the first, second and fifth days after birth, in order to elucidate the relations between neutrophil kinetics and these hematopoietic factors. The G-CSF and GM-CSF receptors on neutrophils were also investigated in 16 healthy newborns. G-CSF and GM-CSF receptor-positive neutrophil percentages were not different from those in the peripheral blood of adult controls. The receptor densities, assessed by mean fluorescence channel number, were also similar in newborn and adult neutrophils. The mean serum concentrations of G-CSF were high (191 pg/ml) on the first day of life and gradually decreased on the second (128 pg/ml) and fifth (112 pg/ml) days. A statistically significant correlation (p < 0.05) was found between G-CSF levels and absolute neutrophil counts (ANC). Furthermore, the percentage of decrease in G-CSF levels correlated significantly with the percentage of decrease in ANC (p < 0.001). GM-CSF levels were also high, though less striking, on the first day of life (9.5 pg/ml), remained at high levels on the second (10 pg/ml), and gradually decreased on the fifth (8.8 pg/ml) day. IL-3 levels were high (110 pg/ml) on the first day of life and remained at high levels on the second (138 pg/ml) and fifth (138 pg/ml) days. We found that the IL-3, GM-CSF and G-CSF levels were elevated during the first week of postnatal life. Our findings suggest that significant changes in the levels of the growth factors are likely to be the cause of significant leukocyte blood picture changes during the first week of life. We found normal GM-CSF and G-CSF receptors in uninfected newborns. PMID- 9339109 TI - The prevalence of factor V Leiden (1691 G-->A) mutation in Turkey. AB - Resistance to activated protein C (APC), which is caused by a single point mutation in the gene for factor V, is a common risk factor for thrombosis. In this study, we screened factor V (FV) Leiden mutation in 81 subjects. The mutation in the heterozygous form was found in 7.1 percent of our normal population. This high frequency suggests that screening for the FV mutation should be considered in patients with a family history of thrombosis. PMID- 9339108 TI - Evaluation of chimerism with DNA polymorphisms in bone marrow transplantation. AB - Evaluation of chimeric status following allogenic BMT is an important tool for monitoring the replacement of host cells with donor cells and for determining the risk of relapse. Polymorphic DNA sequences can be used as powerful markers in identification of donor/recipient genotype differences, even between close relatives. Polymerase chain reaction (PCR) amplification of three variable number of tandem repeat (VNTR) loci and five single-locus polymorphisms (SLP) was used to identify chimerism in 40 recipient-donor pairs. Mixed chimerism was present in 11 patients, and complete chimerism in 29. This PCR method is a rapid and sensitive assay to detect engraftment and evaluate relapse potential, and thus is very useful in the clinical management of BMT patients. PMID- 9339110 TI - Correlation of laboratory and clinical findings with the location of Xp21 deletion in Duchenne muscular dystrophy. AB - Laboratory and clinical features of 28 Duchenne muscular dystrophy patients were evaluated. Positive family history was present in only two cases (7.1%). Dystrophin I-positive fibers were present in 33 percent of the cases with the deletion close to the 5' end of the gene. In the cases with deletion concerning the central part of the gene, all fibers were dystrophin I-negative. In five of the six cases with short stature, the deletion was close to the 5' end of the gene, and short stature was especially seen together with 8th and 13th exon deletion. Statistical analysis concerning the age at which the patient began to have difficulty in standing up and at which he could not walk, did not correlate with the clinical severity and deletion zone, location or extent. PMID- 9339111 TI - Early determination of nutritional problems in pediatric cancer patients. AB - Mild and marginal malnutrition must be identified to prevent the development of severe protein-energy malnutrition in pediatric cancer patients. We aimed to evaluate nutritional status and determine daily energy, protein and micronutrient intake to identify mild or marginal malnutrition in pediatric cancer patients. Daily energy, protein and micronutrient intake, anthropometric measurements and biochemical indices were studied in 45 patients (25 in remission, 20 newly diagnosed or relapsed) who consumed energy, protein, vitamins and minerals below the recommended quantities. According to the weight-for-height index, 23 children (51.1%) were determined to be malnourished. Absolute and relative prealbumin values were 19.4 +/- 7.2 mg/dl and 74.3 +/- 29.1 mg/dl in the remission group, and 14.8 +/- 5.1 mg/dl and 58.1 +/- 23.3 in the active disease group, respectively (p < 0.05). Relative prealbumin values were found to be low in 63.6 percent of nonmalnourished children, and 80 percent of children with mild malnutrition. We conclude that malnutrition is common in pediatric cancer patients, and prealbumin is a reliable and sensitive indicator of mild and marginal malnutrition. Determining prealbumin values and assessing the deficiency of micro- and macronutrients before malnutrition is detected by anthropometric measures may provide a warming that nutritional problems may occur. PMID- 9339112 TI - Renal function in children with hypercalciuria. AB - Hypercalciuria is a common problem causing symptoms such as abdominal pain, hematuria and enuresis, and leading to stone formation. It results from a renal tubular calcium "leak" or intestinal hyper-reabsorption of calcium. This study was performed to determine whether renal functional impairment was present in children with hypercalciuria. The study group comprised 298 children who were screened for hypercalciuria by means of urinary calcium/creatinine (UCa/UCr) ratio. The renal functions of 18 children (6.4%) detected as having hypercalciuria with Ca/Cr ratios of greater than 0.18 in their spot urines were evaluated. Results were compared with those of the healthy control group. The rate of hypercalciuria did not very significantly between the boys and girls (p > 0.05). The mean value of daily calcium excretion was 6.42 + 3.93 mg/kg/day in the children with hypercalciuria, which was significantly different from that of the control group (p < 0.01). When the values of creatinine, osmolar and free water clearances, fractional excretion of sodium and tubular reabsorption of phosphorus were compared between the patient and control groups, the difference was not significant (p > 0.05). Urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion, which was described as the creatinine ratio, was significantly higher in the children with hypercalciuria. These findings suggest that in the presence of normal renal functional studies in children with hypercalciuria, tubular injury can be detected by NAG, which is a more sensitive marker of renal tubular injury. PMID- 9339113 TI - Intravenous immunoglobulin for sepsis prevention in preterm infants. AB - The aim of this study was to investigate whether intravenous immunoglobulin (IVIG) can prevent sepsis in premature newborn infants. The study group consisted of 80 preterm newborn infants, who were divided into two groups: 40 preterm newborns received IVIG prophylactically (group A) and the control group (group B, n = 40) did not receive IVIG. IVIG was given at a dose of 500 mg/kg to infants. weighing greater than 1500 g, and 700 mg/kg to those weighing less than 1500 g at birth on days one, two and eight of life. By two, eight and 12 days of age, the treatment group had significantly greater IgG concentrations than the control group. Mortality was 7.5 percent (3/40) in group A and 27.5 percent (11/40) in group B (p < 0.01). Bacteremia was determined in three blood cultures in group A and eight in group B, particularly S.aureus and S.enteritis. PMID- 9339114 TI - Immunoglobulin isotypes and IgG subclasses in recurrent infections. AB - The frequency of major immunoglobulin isotype and IgG subclass deficiencies among 74 children aged six months to 14 years with recurrent infections was studied. All children had at least five to six episodes of respiratory tract infections, while recurrent diarrhea had occurred in eight. Two selective and six partial IgA deficiencies were detected. IgG4, IgG3 and IgG2 deficiencies, either isolated or combined, were found in 13, nine and one patient respectively. Among these there were two combined deficiencies of IgA + IgG4, three of IgA + IgG3, one of IgA + IgG2 + IgG4, one of IgG1 + IgG3 and two of IgG1 + IgG4. There was one patient with panhypogammaglobulinemia. Our results, similar to those of other studies, showed that the occurrence of the Ig isotype, particularly subclass deficiencies, is not uncommon in children with frequent infections. PMID- 9339115 TI - Management of foreign body aspiration in infancy and childhood. A life threatening problem. AB - It is well known that young children have tendency to place objects in their mouths, frequently leading to aspiration of foreign bodies (FBs) into the tracheobronchial tree (TBT). The patient group comprised 596 patients with a history of suspected aspiration of FBs into the TBT who were bronchoscoped for diagnosis and treatment. There were 306 male (51.3%) and 290 female (48.7%) patients, with a mean age of 2.4 years (range 3 months-13 years). Sunflower seeds and hazelnuts were the most common FBs that were extracted using an open-tube rigid bronchoscope (Storz, Germany) and suitable coaxial forceps (Storz, Germany). Patients admitted within 48 hours following the aspiration numbered 341 (57.2%). The distribution of FBs between the right and left lung and trachea was 53, 37 and five percent, respectively. The aspirated material was visible on the chest x-ray in only 10 percent cases, which facilitated in making the diagnosis. Despite a history of aspiration, bronchoscopy was negative in 21 (3.4%) of the cases. Thoracotomy and subsequent bronchotomy was the treatment of choice in seven (1.5%) and lobectomy in two (0.3%) cases. Cardiorespiratory arrest was observed in five (0.8%) cases, three of whom (0.5%) died during bronchoscopy (2 cases) or thoracotomy (1 case). In conclusion, patients with FB aspiration are rapidly recognized from their histories and easily treated by bronchoscopy and extraction of the aspirated foreign body. A high index of suspicion is crucial for early diagnosis. However, education is the best preventive measure for decreasing the incidence of this problem. PMID- 9339116 TI - Cerebrospinal fluid shunt complications. AB - We report our experience with cerebrospinal fluid shunt procedures performed on 306 patients between 1983 and 1993. Patients were between the ages of one day and 15 years (average 14.9 months) on admission. Three hundred and thirty-six shunt placements and 274 revisions were done. The first complication occurred in the first postoperative month in 52 patients and within the first six months following surgery in 97 patients. Age was determined as a statistically significant factor in only infection and the slit ventricle syndrome (SVS). Shunt types and systems were not significant factors causing complications. The level of consciousness of the patients at the time of surgery influenced the rate of complications; patients with impaired consciousness at the time of surgery had higher complication rates than those operated on in a normal state of consciousness (41% and 8.5%, respectively). PMID- 9339118 TI - Acute pancreatitis in a patient with glutaric acidemia type II. AB - We describe a two-year-old girl who presented with coma following an upper respiratory tract infection. Nonketotic hypoglycemia, metabolic acidosis and mild hyperammonemia were detected. The urinary organic acid profile was consistent with glutaric aciduria type II. Pancreatitis was diagnosed at autopsy. Although pancreatitis has been described in a number of inborn errors of metabolism including organic acidemias, to the best of our knowledge this is the first report of acute pancreatitis occurring in glutaric acidemia type II. It was stressed, therefore, that this complication should be searched for in organic aciduria patients, and the measurement of plasma amylase and lipase levels should be added to the battery of laboratory investigations in such cases. PMID- 9339117 TI - Risk factors associated with corrective surgery in congenital scoliosis with tethered cord. AB - Severe neglected congenital scoliosis with tethered cord presents major difficulties in management. The primary objective of this study was to identify the risk factors associated with corrective scoliosis surgery in neglected cases presenting with severe deformity. Six patients who presented with such problems draw attention to the importance of a staged anterior and posterior approach in order to obtain satisfactory functional results. However, corrective surgery is associated with major neurologic and systemic complications, and every effort should be made to intervene with the progression of the deformity before corrective measures becomes necessary. PMID- 9339119 TI - Leprechaunism in two Turkish patients. AB - We report two cases of Leprechaunism with the classical features. The first case had hyperglycemia and severe hyperinsulinemia. The postmortem examination of the second child revealed enlargement of both ovaries, islet cell hyperplasia in the pancreas, and cholestasis and paucity of bile ducts in the liver. Cystic changes were noted in the ovaries, and the kidneys contained a few small cortical cysts. Both patients died at early ages. PMID- 9339120 TI - IgA nephropathy occurring in two siblings of three families. AB - There have been suggestions in the literature that IgA nephropathy may be familial. Genetic factors may influence the development of disease in an association between HLA antigens and IgA nephropathy. At the present time, no conclusion can be drawn. Here, two siblings with typical IgA nephropathy in three families are presented. A relation between HLA and IgA nephropathy was not detected in these family studies. The first family involved a 14-year-old girl and her brother, who at the age of 12 years were admitted to the hospital with macroscopic hematuria. All of the investigations, including serum IgA levels (104 mg/dl and 102 mg/dl respectively) showed normal kidneys with IgA deposition in the mesangium of the glomeruli. In the second family, a 15-year-old boy and his brother at nine years of age were admitted with macroscopic hematuria and gross hematuria, respectively. Laboratory investigations were normal. The serum IgA level (287 mg/dl) was normal in the first patient but the second was elevated at 485 mg/dl. IgA deposits were observed in the glomerular mesangium in these patients. The third family consisted of a 15-year-old boy and his nine-year-old sister who were both admitted with microscopic hematuria. Serum IgA levels (193 and 131 mg/dl, respectively) and laboratory investigations were normal. Renal biopsy specimens revealed C3 and IgA depositions in the glomerular mesangium of both siblings. In the first family the patients were HLA identical, while in the others the siblings were one-haplotype identical. Although IgA nephropathy and HLA antigens are strongly associated in the literature, we could not find this association. PMID- 9339121 TI - Akinetic mutism due to diphenylhydantoin toxicity. AB - Akinetic mutism (AM) is a rare, specific, unconscious state. An AM patient seems to be awake, lacks mental activity, is unable to speak, and does not respond to any environmental stimulus. Cyclical sleep and awake states are maintained, and incontinence is present. Various factors such as tumor, vascular events, drug use and radiotherapy are responsible for the development of AM. A 12-year-old epileptic patient displayed AM and diphenylhydantoin toxicity (DPH). She seemed awake, was unable to speak or to understand, and had no movements with either spontaneous or noxious stimuli. Her serum DPH level was greater than 40 micrograms/ml. Magnetic resonance imaging (MRI) showed mild cerebellar atrophy. All known causes of AM were excluded. The AM state in this patient was considered to be due to toxic DPH levels. She regained her motor and mental activity within two months after carbamazepine was administered to replace DPH. She was symptom free when examined at the two-year follow-up. No similar adverse effect of DPH has been reported to date. PMID- 9339122 TI - Netherton's syndrome and neonatal hypernatremia. A case report. AB - Netherton's syndrome is characterized by ichthyosiform desquamation, bamboo hair and often atopic diathesis. It is transmitted as an autosomal-recessive trait. In this paper we report a baby with Netherton's syndrome who developed hypernatremia during the neonatal period. This complication should be remembered in erythrodermic infants as a preventable cause of neonatal morbidity. PMID- 9339123 TI - A case of brachyolmia. AB - Brachyolmia refers to a form of skeletal dysplasia characterized by general platyspondyly without significant epiphyseal, metaphyseal or diaphyseal changes in long bones. Three, possibly four, types of brachyolmia have been defined: Type I-Hobaeck-Toledo type. Type II-Maroteaux and Type III. We report a patient with brachyolmia and present the clinical and radiological findings. A 15-year-old boy presented to our Outpatient Department because of his short stature. His height, weight, head circumference and arm span were 127 cm (< 3rd percentile), (3rd percentile) 39 kg, 55 cm (50th-75th percentile), and 142 cm respectively, and his upper segment/lower segment ratio was 0.91. His neck and trunk were short. He had severe kyphoscoliosis. Slit-lamp examination was normal. Radiologic features included platyspondyly in cervical, thoracic and lumbar vertebrae as well as kyphoscoliosis. Bilateral coxa valga and mild acetabular irregularities were noticed on pelvic radiographies. Levels of chondroitin and heparan sulphate as well as the glycosaminoglycan/creatinine ratio were elevated in the 24-hour urine specimen. The activities of N-acetylgalactosamine-6-sulphatase, beta galactosidase and beta-hexosaminosidase were all normal in fibroblast culture. Although the x-ray findings of this patient are consistent with both Types I and III, recessive inheritance and glycosaminoglycan anomalies point to Type I brachyolmia. PMID- 9339124 TI - Radiofrequency catheter ablation treatment of a child with dilated cardiomyopathy secondary to chronic ectopic atrial tachycardia. AB - Ectopic atrial tachycardia (EAT) is a rare but reversible cause of dilated cardiomyopathy (DCMP). The diagnosis and the definite control of the arrhythmia are essential for the regression of DCMP. Unfortunately, conventional antiarrhythmic drugs usually fail to control the arrhythmia, and the results of surgery or direct current ablation are suboptimal. Recently, radiofrequency (RF) catheter ablation has been evolving as a safe and effective therapy for EAT. This report describes the RF ablation treatment of a 14-year-old boy with DCMP secondary to chronic EAT. Activation mapping was used for the purpose of identifying the focus origin located just anterior to the coronary sinus os. RF energy applied at this focus successfully terminated the tachycardia. No complications related to the procedure were observed. RF ablation not only caused elimination of the EAT but also led to improvement in left ventricular function as early as two weeks after the procedure, and complete resolution of DCMP in three months. PMID- 9339125 TI - Bicuspid aortic valve and coarctation of aorta. AB - Bicuspid aortic valve (BAV) and coarctation of aorta (COA) are frequently seen together. It is believed that these malformations result from a single developmental diathesis. A case is presented of COA, aortic stenosis and aneurysm of the ascending aorta corrected by patch aortoplasty and commisurotomy. An aneurysm at the site of the coarctation repair can develop as late as 20 to 25 years after surgery. Almost all of the aneurysms described have occurred in patients undergoing patch aortoplasty. We do not recommend this technique except in special cases. PMID- 9339126 TI - Design of a medical image processing software for clinical-PACS. AB - Software modules for interactive display, manipulation and retrieval of medical images have been designed for a Picture Archiving and Communications System (PACS). The target of these modules is not for a high-end diagnostic workstation for radiologists, but for a PC-based low cost clinical workstation for a referring physician. This software is constructed based on a concept of an object oriented language which is designed to be modular and expandable. It consists of several functional modules: (a) a communication module for image retrieval, (b) a standard module for the interpretation of the DICOM images, (c) a user interface module for the non-computer oriented clinicians and (d) a tool module for viewing and manipulating images as well as editing the annotation. PMID- 9339127 TI - A modified hearing aid fitting procedure using both real ear and 2cc coupler measurement system. AB - In order to reduce the test time in real ear hearing-aid fitting for children, the validity of applying the average real ear to coupler differences (RECDs) in prefitting procedure using a 2cc coupler measurement system was evaluated by checking whether the majority of people's RECDs might occur within 5 dB of the average RECDs (N = 116) in each test frequency and age group. The percentages of occurrence were around 90% in test subjects' RECDs in saturation sound pressure levels (SSPLs) and around 70% in gain in each important test frequency. Appropriate test frequencies in prefitting are 500, 1000, 1500 and 2000 Hz. PMID- 9339130 TI - A path analysis on prisoners' health behavior and medical utilization. AB - In this thesis, Korean prisoners' health behavior and the characteristics of their medical utilization were surveyed and analysed. Because prisoners are inclined to be mediators of communicable diseases or unhealthy behaviors between prison institution and the outside world, health care for prisoners is directly related to the national population. Data were collected through a self administered survey of 5 Korean prisons out of a total of 38 correctional facilities and analysed in accordance with a causal model based on a path frame, by serial multiple regressions on health behavior, health status, and medical utilization, etc. According to the survey analysis, while prisoners were generally concerned with their health much more than they were before imprisonment, they perceived that their health status had deteriorated after imprisonment, and that their need for health services was increasing gradually during their time in prison. In the path analysis on the causal relations among variables related to the prisoners' health status and medical utilization, the prisoners' characteristics affected their health concern and health behavior, and subsequently affected their health status and medical utilization, respectively. To sum up these exploratory studies on prisoners' health behavior and health service utilization, some efforts to organize a health care system embracing the correctional institution and health care administration should be made on the level of establishing a health care delivery system for special social groups like prisoners. PMID- 9339129 TI - Monitoring of WT-1 gene expression in peripheral blood of patients with acute leukemia by semiquantitative RT-PCR; possible marker for detection of minimal residual leukemia. AB - The expression of the WT-1 gene which is found exclusively in human leukemic blasts frequently disappears from bone marrow of leukemia patients in complete remission (CR). Using semiquantitative RT-PCR, we investigated the expression of the WT-1 gene in peripheral bloods (PBs) of 33 patients with acute leukemia (AML 26; ALL 7) and monitored its expression after achievement of CR. None of the 6 normal controls expressed detectable levels of WT-1 transcripts (< 10(-4), background level), whereas 31 (93.9%) of 33 patients expressed variable levels of WT-1 transcripts (range, 10(-4) to 10(1)) at diagnosis. The level of WT-1 expression was not different between AML and ALL. By monitoring WT-1 gene expression in PB of 31 patients during CR, 5 patients relapsed (two from the 18 patients with undetectable levels of WT-1 gene expression and three from the 13 with WT-1 gene expression in low levels). Three of the 5 relapsed patients showed preceding reappearance or rise of WT-1 gene expression. From these results, we reconfirmed that the WT-1 gene is a pan-acute leukemic marker, which can be used to monitor minimal residual leukemia (MRL) after chemotherapy or in patients with CR. PMID- 9339128 TI - Correlations of bcl-2 expression with clinicopathological features in breast cancer. AB - To evaluate the prognostic significance of bcl-2, we investigated the correlation of bcl-2 expression with the established indicators of prognosis and tumor behavior in breast cancer. This study included a patient group of 91 histologically diagnosed female breast carcinomas. To determine the bcl-2 immunoreactivity, we used a monoclonal antibody directed against the bcl-2 protein by immunohistochemistry from paraffin-embedded tissue in a series of 91 women with breast cancer. Interpretable DNA histograms were obtained from 84 patients. The median age at diagnosis was 45.5 years and the median follow-up time was 30.5 months. Forty-eight (52.7%) cancers showed the bcl-2 immunoreactivity in the cytoplasm. The nonneoplastic portion of ductal epithelial cells and normal lymphocytes were usually stained with bcl-2 antibody. Estrogen receptors (ER)(p < 0.001) and progesterone receptors (PR)(p < 0.001) showed strong positive correlation with bcl-2 immunoreactivity. The histologic grade (p < 0.05) and nuclear grade (p < 0.01) also showed positive relationships with bcl 2 positivity but tumor size (p > 0.05) and DNA ploidy (p > 0.05) were not related with it. The bcl-2 positive patients showed longer survival (p < 0.05) compared to bcl-2 negative tumors in univariate analysis (Kaplan-Meier life table analysis). Using multivariate analysis with Cox regression, bcl-2 (p > 0.05), nuclear grade (p > 0.05), ER status (p > 0.1) and PR status(p > 0.1) were not reliable indicators for overall survival except histologic grade (p < 0.05). Our results suggest that bcl-2 expression may be related to hormonal regulation and tumor differentiation in breast carcinoma. Larger patient study groups with a longer follow-up period will be helpful to clarify the prognostic significance of bcl-2. PMID- 9339131 TI - Cold haemagglutinin disease in systemic lupus erythematosus. AB - A 34-year-old lady presenting with features of cold agglutinin disease during the course of systemic lupus erythematosus is described. Cold antibody titer was very high (1 in 4096) with specificity for 'I' antigen. Even though she had poor prognostic factors like high titer of cold antibodies with low thermal amplitude, she responded well to prednisolone. PMID- 9339132 TI - Acute monoarthritis associated with positively birefringent maltese cross appearing lipid spherules in a hyperlipidemic diabetic patient. AB - A 63-year old man developed acute monoarthritis in the dorsum of the left foot. Polarized light microscopy of the synovial fluid from his third metatarsophalangeal joint revealed numerous positively birefringent lipid spherules with a maltese cross appearance. Positively birefringent lipid spherules can be found in association with acute, otherwise unexplained arthritis, and may induce synovial inflammation similar to that seen in other types of crystal-induced arthritis. We report a case of acute monoarthritis in which large numbers of positively birefringent lipid spherules were present in a hyperlipidemic diabetic patient. PMID- 9339133 TI - A case of bleeding from the Dieulafoy lesion of the jejunum. AB - Dieulafoy lesion is an uncommon cause of gastrointestinal bleeding, reported to be only 2% of acute or chronic upper gastrointestinal bleeding episodes. Bleeding occurs from a small mucosal erosion involving an unusually large submucosal artery in an otherwise normal mucosa. It is associated with massive, life threatening hemorrhage and is difficult to diagnosis. In most cases the lesion is encountered in the proximal stomach, antrum, duodenum, colon and rectum. In particular, extragastric Dieulafoy lesion is an extremely rare source of intestinal bleeding. In Korea, no case of bleeding from a Dieulafoy lesion of the small intestine has been previously reported. We experienced one case of bleeding from a jejunal Dieulafoy lesion, which was confirmed by the pathologic examination of the resected specimen, and report here. PMID- 9339134 TI - EBV-elicited familial hemophagocytic lymphohistiocytosis. AB - Familial hemophagocytic lymphohistiocytosis (FHL) is a rapidly fatal illness, usually encountered in infancy, characterized by fever, hepatosplenomegaly, pancytopenia, and central nervous system involvement. Microscopic examination of tissue shows a non-malignant lymphohistiocytic infiltrate, with prominent erythrophagocytosis. FHL is an autosomal recessive hereditary disorder but may develop secondarily to other conditions such as immunosuppression, malignancies, fat overload and certain infections. We recently experienced a case of siblings developing FHL, which may be associated with EBV infection. PMID- 9339135 TI - Congenital bronchoesophageal fistula associated with esophageal diverticulum in the adult. AB - Congenital bronchoesophageal fistula is a rare clinical entity in adults. This anomaly may cause various symptoms such as respiratory infections, coughing bouts when eating or drinking, and even hemoptysis. The fistula can cause symptoms in childhood but may not appear until adulthood. We recently experienced a case of congenital bronchoesophageal fistula associated with esophageal diverticulum in an adult. A 63-year-old woman was admitted to our hospital due to chest discomfort, sore throat and coughing bouts when eating. An empyema with lung abscess had occurred eight years previously. Results of the physical examination were unremarkable. A Barium swallowing revealed a medium-sized diverticulum at the right anterior aspect of the esophagus, which had developed a fistulous connection with the right lower lobe bronchus. The patient was treated by fistulectomy and lobectomy of the right lower lobe. The postoperative course was smooth and uneventful. PMID- 9339136 TI - The Alan Welford Memorial Lecture. Ageing and human skill: a 40th anniversary. AB - Even 40 years after it was published, a re-reading of Alan Welford's seminal book, Ageing and Human Skill, offers important and curiously neglected ideas. In particular, it shows that current 'single factor' models for cognitive ageing, and for general intellectual abilities neglect the complexity of even apparently very simple tasks; that older individuals can be impressively good at acquiring new skills; that the performance of older people on novel tasks is different from that of young adults not only in quantitative terms, because they are slower and less accurate, but also qualitatively, in ways similar to individuals who have suffered damage to the frontal lobes of their brains. We celebrate Alan's prescience in being the first to clearly raise these issues, and to discuss elegant but sadly neglected experimental paradigms and techniques of analysis that allow them to be explored. PMID- 9339137 TI - Ergonomics, quality and continuous improvement--conceptual and empirical relationships in an industrial context. AB - This paper reviews the literature comparing the fields of ergonomics and quality, mainly in an industrial context, including mutual influences, similarities and differences. Relationships between ergonomics and the factors: work conditions, product design, ISO 9000, continuous improvements and TQM are reviewed in relation to the consequence, application, and process domains. The definitions of ergonomics and quality overlap substantially. Quality deficiencies, human errors and ergonomics problems often have the same cause, which in many cases can be traced to the design of work, workplace and environment e.g. noise, light, postures, loads, pace and work content. In addition, the possibility of performing to a high standard at work is an important prerequisite for satisfaction and well-being. Contradictions between the two fields have been identified in the view of concepts such as standardization, reduction of variability and copying of best practice, requiring further research. The field of quality would gain by incorporating ergonomics knowledge, especially in the areas of work design and human capability, since these factors are decisive for human performance and also therefore the performance of the systems involved. The field of ergonomics, on the other hand, would benefit from developing a stronger emphasis on methodologies and structures for improvement processes, including a clearer link with leadership and company strategies. Just as important is a further development of practicable participative ergonomics methods and tools for use at workplaces by the workers themselves, in order to integrate the top-down and the bottom-up processes and achieve better impact. Using participative processes for problem-solving and continuous improvement, focusing ergonomics and quality jointly has a great potential for improving working conditions and quality results simultaneously, and satisfying most of the interested parties. PMID- 9339138 TI - Psychosocial aspects of working with video display terminals (VDTs) and employee physical and mental health. AB - Psychosocial aspects of using video display terminals (VDTs) have been recognized as contributors to employees' mental and physical health problems for more than 15 years. Yet, little has been done by employers to change work organization conditions to improve the psychosocial work environment of VDT users. Thus, psychosocial aspects of work are emerging as one of the biggest problems for VDT users in the late 1990s. This paper explores how psychosocial aspects of VDT work are related to job stress, and their consequences for mental and physical health. Using the research literature, it defines various aspects of work organization and job design that have been shown to be related to VDT users' ill-health. Some of the important work design aspects uncovered include a lack of employee skill use, monotonous tasks, high job demands and work pressure, a lack of control over the job, poor supervisory relations, fear of job loss, and unreliable technology. These are the same job stressors that have been defined as problematic for a variety of blue collar jobs in previous research. Work organization improvements for healthier VDT jobs are proposed. These include organizational support, employee participation, improved task content, increased job control, reasonable production standards, career development, enhanced peer socialization, and improved workstation ergonomics. These organizational improvements are derived from a more detailed organizational strategy for job stress reduction. A model of job redesign through proper 'balancing' of work organization features is discussed. PMID- 9339139 TI - Human factors in modern traffic systems. AB - Traffic systems are undergoing enormous change with the advent of Intelligent Transport Systems (ITS). Although productivity and quality of mobility are emerging interests, safety remains the predominant preoccupation of ITS human factors. It should be evident that while intelligent technologies may have the potential to improve traffic safety, they also have the potential to adversely affect it. Ultimately, the effect on safety depends on the specific technologies that are invoked and the manner in which they are incorporated within the vehicle as well as within the larger road transportation system. Current automotive developments can be characterized as technology-centred solutions rather than user-centred solutions. Greater effort must be directed at understanding and accommodating the human element in the road transportation system in order that future transportation objectives can be achieved. There is a need to expand the scope of traditional human factors to include macro-level effects as well as to place greater emphasis on understanding human interactions with other elements of the system. There is also increasing recognition of the urgent need for systematic procedures and criteria for testing the safety of ITS prior to large scale market penetration. PMID- 9339140 TI - History and future of ergonomics in building and construction. AB - In this paper an overview is given of what has been done in the field of ergonomics on behalf of the building and construction industry. Characteristics of the building and construction are given in relation to working conditions. The developments within this branch of industry are described in view of perspectives for a better future. PMID- 9339141 TI - Manual handling of loads: the point of view of experts involved in the application of EC Directive 90/269. AB - This paper illustrates the viewpoint of a group of Italian research workers who have been directly involved in providing criteria and guidelines for application of EC Directive 90/269, Manual handling of loads. The problems posed by application of the Directive in Italy and which are mainly related to exposure assessment, health surveillance and risk management (work and workplace redesign) are thoroughly examined. Major questions still to be solved include defining methods for assessment of multiple and complex load handling tasks, identifying data sources on 'health effect' trends as investigated in a working population with low exposure or in the general population in order to fix action levels, and setting up a data bank of technical and organizational solutions and products accessible to experts and users. PMID- 9339142 TI - The scientific basis for making guidelines and standards to prevent work-related musculoskeletal disorders. AB - Regulations concerning the work environment, tools, and the performance of work are at their best based on scientific evidence. Existing European directives, European and North American standards, and recent guidelines with the potential to prevent musculoskeletal disorders, are either qualitative or semiquantitative. The exception is the NIOSH lifting guide, which is highly quantitative. Of the European directives and standards, few have been developed with the primary goal of preventing musculoskeletal disorders, whereas one North American standard and another suggestion for a standard have this specific aim. In a review of epidemiological studies on low-back, neck, shoulder, and upper extremity disorders, several physical load factors were identified as risk factors for the disorders. Many of these factors have been repeatedly identified, and for different types of outcomes of an anatomical area (e.g. pain, disc herniation, disc degeneration of the low-back or neck). However, quantitative exposure response relationships between physical load factors and disorders based on field studies are largely unknown. Experimental studies have provided a multitude of potentially useful data. It is concluded that both well-designed epidemiological studies with quantitative assessments of physical work load and valid measurements of musculoskeletal disorders, and experimental studies are needed for the future development of regulation. To determine the role of experimental studies in regulation, it should be known to what extent fatigue and other short term responses are precursors of disorders. Regulation should be directed especially towards factors that are likely to be causative for musculoskeletal disorders. Examples of such factors are sudden overload in manual handling activities, heavy physical work involving manual handling tasks, and vibration from tools. Guidelines that are acceptable and feasible can and should be developed. The effects of such guidelines on the occurrence of musculoskeletal disorders should be investigated. PMID- 9339143 TI - Ergonomics and safety in societies in transfer. AB - In Central Europe, the influence of transformation on science and practice in both ergonomics and occupational safety has been positive. The opening of markets has automatically resulted in the quality of products of various countries being compared. The comparison of the state of science has been equally revealing. The spontaneous willingness of leading world centres to co-operate in both occupational safety and ergonomics has resulted in positive changes, e.g. intensive work on creating the instruments for: implementing ILO conventions and EU directives into national laws; implementing international and European standards into national standards; accrediting testing laboratories in the field of occupational safety and ergonomics; accrediting centres for product certification for the safety mark (obligatory) and for conformity with ergonomic parameters (voluntary); and computer-aided designing and creating databases in occupational safety and ergonomics conforming to international standards. These are the laws of the emerging common market for products and services. There is still a much more difficult area of necessary changes in the approach to: the value of life and health; the belief in the possibilities and the effectiveness of initiatives towards changing the working and life environment; and the form and content of the information in occupational safety and ergonomics taught from school to adult education. Transformation has led to a renaissance in which man has become the subject of all aspects of life and activity. There is also a renaissance of occupational safety and ergonomics. The fields of research that have gained importance in this new approach in Central Europe are discussed. PMID- 9339146 TI - 40th Annual meeting of the GTH (Gesellschaft fur Thrombose- und Hamostaseforschung). Interlaken, Switzerland, 14-17 February 1996. Abstracts. PMID- 9339147 TI - American College of Rheumatology 61st National Scientific Meeting and Association of Rheumatology Health Professionals 32nd National Scientific Meeting. Washington, DC, November 8-12, 1997. Abstracts. PMID- 9339148 TI - [The "STOP NIDDM" program--can diabetes in the aged be prevented? An international long-term study revisited; does Acarbose delay or prevent the manifestations of type II diabetes]. PMID- 9339149 TI - 1996 Orthopaedic proceedings. PMID- 9339150 TI - NKF-DOQI clinical practice guidelines for vascular access. National Kidney Foundation-Dialysis Outcomes Quality Initiative. PMID- 9339151 TI - NKF-DOQI clinical practice guidelines for the treatment of anemia of chronic renal failure. National Kidney Foundation-Dialysis Outcomes Quality Initiative. PMID- 9339152 TI - Pregnancy after assisted ejaculation procedures in men with spinal cord injury. AB - OBJECTIVE: To present the results of fertility treatment in 28 men with spinal cord injury (SCI) and their partners. DESIGN: Retrospective analysis. SETTING: University hospital outpatient clinic and home. PATIENTS: Twenty-eight anejaculatory men with SCI and their partners seeking treatment for infertility. INTERVENTION: Penile vibratory stimulation and electroejaculation as semen retrieval methods. Assisted reproductive techniques used: vaginal self insemination at home, intrauterine insemination, in vitro fertilization with or without intracytoplasmic sperm injection. MAIN OUTCOME MEASURES: Ejaculation rate; sperm count and motility; pregnancy rates. RESULTS: All of the men were able to ejaculate either by penile vibratory stimulation (79%) or electroejaculation (21%). Median total sperm count was 65 million (range, 0.1 to 480) with a median motility of 13% (range, 1% to 60%). Overall, 9 of 28 couples (32%) achieved 10 pregnancies (4 self-insemination, 3 intrauterine insemination, 1 in vitro fertilization, and 2 intracytoplasmic sperm injection). CONCLUSIONS: An ejaculation rate of 100% was achieved using penile vibratory stimulation as a first treatment option with electroejaculation as a second option. Motivated couples with adequate semen quality may be offered penile vibratory stimulation combined with self-insemination at home. Together with intrauterine insemination or fertilization techniques used in vitro, the pregnancy rate per treatment cycle for SCI couples may approach that of natural procreation in healthy and fertile couples. PMID- 9339153 TI - Pharmacologically initiated defecation for persons with spinal cord injury: effectiveness of three agents. AB - OBJECTIVE: To compare the effectiveness of hydrogenated vegetable oil-based bisacodyl (HVB) suppositories, polyethylene glycol-based bisacodyl (PGB) suppositories, and polyethylene glycol-based, glycerine, docusate sodium mini enemas (TVC) in subjects with upper motor neuron spinal cord lesions. STUDY DESIGN: Prospective randomized double blind. Fifteen subjects received one of 3 HVB and 3 PGB suppositories in randomized sequence for each of six scheduled bowel care sessions. Additionally, 10 subjects received 3 TVC. The analysis used timed events that divided the bowel care sessions into discrete intervals. The analysis also compared digital simulations, incontinence, and quantity of stool. Wilcoxon rank sum tests and paired t tests were used to compare the means of intervals during bowel care initiated by HVB, PGB, and TVC. RESULTS: (means in minutes and p values): Time to Flatus-HVB, 32; PGB, 15; TVC, 15; p < .026, HVB PGB; p < .983, PGB-TVC; Flatus to Stool Flow-HVB, 6.7; PGB, 5.5; TVC, 3.9; p < .672, HVB-PGB; p < .068, PGB-TVC; Defecation Period-HVB, 36; PGB, 20; TVC, 17; p < .037, HVB-PGB; p < .479, PGB-TVC; Wait Until Transfer-HVB, 10.9; PGB, 10.7; TVC, 7.4; p < .932, HVB-PGB; p < .043, PGB-TVC; Total Time for the bowel program HVB, 74.5; PGB, 43; TVC, 37; p < .010, HVB-PGB; p < .458, PGB-TVC; percent incidence of incontinence between bowel care sessions-HVB, .067; PGB, .067; TVC, .033; p < 1.0, HVB-PGB; p < .678, PGB-TVC; amount of stool produced-HVB, 3.30; PGB, 3.49; TVC, 3.38; p < .276, HVB-PGB; p < .630, PGB-TVC; average number of digital stimulations per bowel care procedure-HVB, 4.4; PGB, 4.1; TVC, 3.8; p < .411, HVB-PGB; p < .293, PGB-TVC; time per digital stimulation in seconds-HVB, 107; PGB, 40; TVC, 83; p < .149, HVB-PGB; p < .352, PGB-TVC; and the total time, in minutes, spent performing digital stimulations during bowel care-HVB, 10.0; PGB, 2.7; TVC, 5.9; p < .151, HVB-PGB; p < .325, PGB-TVC. CONCLUSION: Bowel care took less time when initiated with the PGB bisacodyl suppository or TVC mini enema as compared with the HVB bisacodyl suppository (p < .01). PMID- 9339154 TI - Lycra garments designed for patients with upper limb spasticity: mechanical effects in normal subjects. AB - OBJECTIVE: To assess the stretch of pronator muscles produced by a specifically designed upper-limb Lycra garment that could have a better acceptability than rigid splints in treating upper-limb spasticity. DESIGN: Double-blind comparison among three garments. They were designed to produce a supinating, a pronating, and no torsional force, and were individually manufactured and tested in 10 healthy volunteers. MAIN OUTCOME MEASURE: Angular position and passive rotational stiffness of the forearm were measured with and without each of the garments immediately after the garment was fitted and every hour for 6 hours. RESULTS: When put on by a trained person, the supinator garment supinated the forearm in all subjects (mean, 17 degrees; p < .01; range, 5 degrees to 44 degrees) while the pronator garment pronated the forearm in 8 of 10 subjects (mean, 5 degrees; p < .01). These effects gradually decayed over 6 hours, as garment position was not readjusted. Passive rotational stiffness of the forearm increased by about 30% with each type of garment. The garments designed to produce no torsional force exerted no intrinsic rotational effect. CONCLUSION: Individually made Lycra garments can produce continuous stretch of muscles for several hours and may be useful in the treatment of spasticity. The garments, however, must be put on by a trained person and their position adjusted when necessary. PMID- 9339155 TI - Postural responses to unilateral arm perturbation in young, elderly, and hemiplegic subjects. AB - OBJECTIVE: To compare postural responses during standing associated with self paced unilateral arm flexion exhibited by young and elderly healthy subjects and subjects with hemiplegia from stroke. DESIGN AND SETTING: Descriptive cross sectional study in a laboratory setting. PATIENTS: Ten young, 10 elderly healthy subjects, and 12 volunteers with longstanding hemiparesis following stroke were tested. The hemiplegic group had good functional balance scores on the Berg Balance Scale (BBS). MAIN OUTCOME MEASURES: The peak arm acceleration, center of pressure (CP) excursion speed, and the electromyographic activity in the posterior leg muscles (bilateral hamstrings and triceps surae) were monitored during self-initiated rapid unilateral arm flexion and during quiet stance. RESULTS: During unilateral arm flexion, the young group showed the highest arm accelerations and lowest CP excursion speeds. The variability of postural responses was largest in the elderly and hemiplegic groups with hemiplegic subjects showing the greatest CP excursion speeds for the lowest arm accelerations. The first burst of activity in ipsilateral hamstrings muscle was the same in all subjects. However, the hemiplegic group had less activation (latency and amplitude) of other posterior leg muscles. CONCLUSION: The elderly subjects had more variable responses to perturbation than the young subjects despite similar BBS scores. This may indicate that the BBS fails to detect differences in balance at the high end of the scale. Although the hemiplegic subjects demonstrated some anticipatory control of standing balance, they consistently performed poorer than elderly and young controls. The failure to coordinate postural leg muscle activity with focal movement may contribute to the instability of subjects with hemiplegia. PMID- 9339156 TI - Generalizability of the limits of stability test in the evaluation of dynamic balance among older adults. AB - OBJECTIVE: Reliability of platform posturography tests is essential for the identification and treatment of balance-related disorders. The purposes of this study were to establish the reliability of the limits of stability (LOS) test and to determine the relative variance contributions from identified sources of measurement error. DESIGN: Generalizability theory was used to calculate (1) variance estimates and percentage of variation for the sources of measurement error, and (2) generalizability coefficients. Random effects repeated measures analysis of variance (RM ANOVA) was used to assess consistency of measurements across both days and targets. PARTICIPANTS: Thirty-eight community-dwelling older adults with no recent history of falls. MAIN OUTCOME MEASURES: Outcome measures derived from the LOS tests included movement velocity (MV), maximum center of gravity (COG) excursion (ME), end point COG excursion (EE), and directional control (DC). RESULTS: Estimated generalizability coefficients for 2 and 3 days of testing ranged from .69 to .91. Relative contributions of the day facet were minimal. The RM ANOVA results indicated that for three of the movement variables, no significant differences in scores were observed across days. CONCLUSIONS: The 75% and 100% LOS tests are reliable tests of dynamic balance when administered to healthy older adults with no recent history of falls. Dynamic balance measures were generally consistent across multiple evaluations. PMID- 9339158 TI - Bilaterally decreased motor performance of arms in patients with chronic tennis elbow. AB - OBJECTIVE: To measure the motor performance of arms in patients with chronic unilateral tennis elbow. DESIGN: Cross-sectional case-control study. SETTING: University hospital clinic admitting chronic hand patients. SUBJECTS: Thirty-two patients with chronic unilateral tennis elbow syndrome and 32 age- and gender matched controls. MAIN OUTCOME MEASURES: The motor performance of arms was measured with the Human Performance Measurement/Basic Elements of Performance system using the module for hands and the protocol of the device. Reaction times, speed of movement, and coordination as a combination of speed of movement and accuracy (number of correct hits) were measured. The results were compared between patients and controls. RESULTS: Simple one-choice and two-choice reaction times were 19% to 36% slower in the patients than in the controls, and speed of movement was 31% to 32% slower in the patients than in the controls. The differences were statistically significant. The coordination results were 9.6bits/sec in the patients and 9.7bits/sec in the controls. The difference was not statistically significant. The reaction times and speed of movement did not differ significantly between the patients' involved and healthy arms. The patients' healthy arms showed significantly slower reaction times and speed of movement than the corresponding arms of the controls. CONCLUSIONS: Unilateral chronic tennis elbow patients have bilaterally decreased reaction times and speed of movement of arms compared with age- and gender-matched controls. The cause for this phenomenon is unclear; the decreased motor performance may be primary and show an increased susceptibility to develop the tennis elbow syndrome or it may be a result of chronicity. PMID- 9339157 TI - Biomechanic effects of a contralateral shoe-lift on walking with an immobilized knee. AB - OBJECTIVES: A previous study demonstrated that when one knee is artificially immobilized, a contralateral shoe-lift improves the oxygen cost of walking. This study was undertaken to evaluate the kinematic and kinetic effects associated with this shoe-lift. DESIGN: Motion analysis and force platform data were collected in subjects walking (1) normally, (2) with one knee immobilized, (3) with one knee immobilized and with a one-half-inch shoe-lift applied to the contralateral, nonimmobilized shoe, and (4) with a one-inch shoe-lift similarly applied. Kinematic and kinetic data from three trials of each condition were compared graphically and statistically using a repeated measures analysis of variance. SETTING: A gait laboratory. SUBJECTS: Eight able-bodied subjects without known neurologic or musculoskeletal problems. MAIN OUTCOME MEASURES: Fifty-two peak kinematic and kinetic variables during various phases of the gait cycle. RESULTS: Statistically significant differences (p < .05) between the normal and immobilized knee conditions were noted in 22 variables; however, significant differences between the immobilized knee conditions were found in only 4 variables. There were small improvements with the shoe-lifts toward normal in peak hip abduction, hip abduction at 20% to 30% of the gait cycle, and in peak knee extension moment on the nonimmobilized side. There was a small change away from normal in peak knee extension moment on the immobilized-side for the 1" shoe lift. CONCLUSION: Wearing a contralateral shoe-lift when one knee is immobilized is associated with only small changes in kinematic and kinetic parameters. The shoe-lift may slightly reduce the need for compensatory hip abduction and vaulting on the nonimmobilized side. Importantly, no adverse biomechanic effect from the shoe-lift was noted except for a slightly increased peak knee extension moment on the immobilized side found for the 1" but not the 1"/2 shoe-lift. PMID- 9339159 TI - Written language production and neuropsychological function in children with traumatic brain injury. AB - OBJECTIVE: To establish the early consequences of traumatic brain injury (TBI) on spontaneous written language production in children by examining writing deficits as a function of injury severity and correlating written performance with neuropsychological data. DESIGN: Case-control cohort study. SETTING: Two regional medical centers. PATIENTS: Seventy-six children, aged 8 to 15 years, with mild, moderate, or severe closed head injury were compared with controls who were individually matched on the premorbid characteristics of age, gender, school grade, behavior, and academic performance. MAIN OUTCOME MEASURES: Assessment of written language production and neuropsychological function was undertaken approximately 1 month following resolution of posttraumatic amnesia. Case-control differences on 16 measures of spontaneous written narratives were computed. RESULTS: Factor analysis and conceptual similarities suggested that the measures of written language fell into five domains: Efficiency, Completeness, General Readability, Error, and Vocabulary. A highly significant, moderate correlation between these measures and severity of injury was found. The highest correlations were found for the written language domains of Efficiency and Completeness and the lowest for the Vocabulary domain. Moderate correlations were also found between measures of written language and neuropsychological function. CONCLUSIONS: At 1 month after resolution of posttraumatic amnesia, written language production in children with TBI is negatively correlated with severity of injury. Some aspects of written language production are affected to a greater extent than others. Considerable common ground was found between written language production and neuropsychological function. PMID- 9339160 TI - Posttraumatic amnesia: its relation to functional outcome. AB - OBJECTIVE: To investigate the relation between duration of posttraumatic amnesia (PTA) and functional outcome in a traumatically brain injured population. PATIENTS: Two hundred seventy-six patients with traumatic brain injury (TBI) who were admitted to a Level I university trauma center and required inpatient rehabilitation. MEASURES: Duration of PTA was assessed by serial administrations of the Galveston Orientation Amnesia Test (GOAT). Functional Independence Measure (FIM) total scores, FIM cognitive and motor subscores, and Disability Rating Scale (DRS) scores were obtained at admission and discharge from inpatient rehabilitation. RESULTS: Duration of PTA was a significant predictor of all admission and discharge DRS and FIM scores. Duration of PTA and age at the time of injury, in combination, contributed significantly to the prediction of the DRS score and FIM total, cognitive, and motor scores at discharge. CONCLUSION: Duration of PTA appears to be a useful variable in predicting specific functional outcome in the TBI population receiving inpatient rehabilitation services. The use of age as a factor in addition to duration of PTA enhances the prediction of functional outcome. PMID- 9339161 TI - Changes in postural sway and performance of functional tasks during rehabilitation after traumatic brain injury. AB - OBJECTIVE: To investigate changes in postural sway while standing, walking parameters, and performance of functional tasks during rehabilitation in a group of traumatic brain injury (TBI) patients. DESIGN: Descriptive. SETTING: Inpatient brain injury rehabilitation unit. PARTICIPANTS: Thirteen subjects undergoing rehabilitation after severe TBI. OUTCOME MEASURES: Two assessments were performed, 2 to 6 weeks apart that included the following: postural sway in three stance conditions; temporal and spatial parameters of walking; functional assessments of walking, standing up, reaching while standing, and stair climbing. RESULTS: There were significant reductions in postural sway in all stance conditions (p < .05) and significant increases in velocity of walking (p < .05), stride length (p < .01), and left and right step lengths (p < .01). Performance on all functional tasks improved (p < .05) except for functional reach. There were no significant correlations between changes in postural sway and changes in walking parameters or functional assessments. CONCLUSION: This study demonstrated significant improvements in postural sway, walking parameters, and functional tasks during a relatively short period of rehabilitation after severe TBI. Improvements in standing balance appear to be independent of improvements in walking performance, suggesting that different mechanisms underlie improved control of these tasks. PMID- 9339162 TI - Review criteria for stroke rehabilitation outcomes. AB - OBJECTIVES: To develop review criteria from the Agency for Health Care Policy and Research Stroke Rehabilitation Guidelines, to review chart records from three sites of care, and to evaluate the interrater and intrarater reliability for the chart review. DESIGN: A descriptive cross-sectional study using a convenience sample. SETTING: Charts for abstraction were obtained from three sites of care home health care, nursing facilities, and inpatient rehabilitation centers. PARTICIPANTS: Charts were included in the study from the three sites of care if the following conditions were met: (1) the client's first admission to a rehabilitation setting; (2) the client's care was Medicare reimbursed; (3) the client lived in the community prior to the stroke; and (4) the client was receiving skilled rehabilitation services. MEASURES: Review criteria, developed directly from the AHCPR Stroke Rehabilitation guidelines, consisted of 11 global quality criteria representative of comprehensive multidisciplinary rehabilitative care. There were approximately 150 variables, comprised of specific criteria to measure each of the 11 global quality criteria plus comprehensive demographic and client-specific information. RESULTS: Results of this study suggest that differences exist in documentation of care across the three sites of care. There was difficulty in obtaining adequate numbers of home health charts. Intrarater reliability, using Cohen's Kappa, was .78 and interrater reliability was .64. CONCLUSIONS: Based on chart documentation, there is variability in the process of stroke rehabilitation care across nursing facilities, inpatient rehabilitation facilities, and home health. This variability can be reliably assessed by chart review. This study provides the impetus for future research specifically evaluating the associations between documentation of the processes of care and patient outcomes. PMID- 9339163 TI - Characterization of global synkineses during hand grip in hemiparetic patients. AB - OBJECTIVE: Global synkineses are defined as nonpurposive associated movements on the involved side of hemiparetic subjects that are triggered during a voluntary movement. The purpose of this study was to characterize the intensity and pattern of upper limb global synkineses in hemiparetic subjects with a static biarticular dynamometer and electromyography during maximal progressive hand grip on the unaffected side. DESIGN: Survey, convenience sample. SETTINGS: University secondary care rehabilitation center. DATA SET: Global synkineses (ie, torques and electromyographic activities) in patients with severe (n = 8) and moderate (n = 7) deficits in motor performance, as evaluated by the Fugl-Meyer assessment, were compared with those obtained in a group of healthy subjects (n = 11). Clinically the subjects from the severe deficit group were more spastic and showed less strength at the elbow than the subjects from the moderate deficit group. RESULTS: Results of analyses of variance showed significant increases of shoulder torque in flexion and internal rotation, and elbow torque in flexion, with increasing force exertion during contralateral hand grip in subjects with severe deficits (p < .05). Furthermore, in these subjects increases of electromyographic activity were also observed in biceps brachii, brachioradialis, and triceps brachii muscles with increasing hand grip force levels. In contrast, no significant torques or electromyographic increases were observed in subjects with moderate deficits and in control subjects during contralateral hand grip exertions. CONCLUSION: These results provide a quantitative assessment of the kinematic and electromyographic patterns of global synkineses and their correlates with clinical observations. Within the limits of the experimental results presented in this study, it is suggested that global synkineses result from contralateral overflow of the voluntary command to hyperexcitable motoneuron pools. PMID- 9339164 TI - Somatosensory evoked potentials of the posterior tibial nerve in hemiparetic patients: relation to stance balance and walking ability. AB - OBJECTIVES: To examine the association between stance ability and walking performance of poststroke hemiplegic patients and their posterior tibial nerve somatosensory evoked potentials (SEPs). DESIGN AND SETTING: Fifteen patients, residents of a geriatric rehabilitation hospital, were evaluated twice, with a 2 week interval between sessions. In each session, clinical tests of stance balance and walking ability were performed, and bilateral SEPs to stimulation of the posterior tibial nerve were recorded. Eight healthy, age-matched control subjects underwent the same tests in a single session, but SEPs were recorded unilaterally. Correlation analysis and analysis of variance (ANOVA) were used for studying the prognostic value of the initial posterior tibial nerve SEP measurements as well as the within- and between-sessions relationships between the clinical-functional tests and the SEP data. RESULTS: No significant correlations between the initial SEP values and functional improvement were established. Within each session, positive significant correlations existed between decreased latencies of several of the medium-latency SEP waves and the performance of stance and gait tasks. However, the between-sessions improvement in stance balance was not correlated with a decrease in latency of the SEP peaks or with an increase in their amplitudes. As to walking ability, in those patients whose gait significantly improved, a significant shortening of P37 and P54 latencies took place. CONCLUSIONS: The association between the initial and/or the 2-week changes in SEP of the posterior tibial nerve and improvement in stance and walking abilities is equivocal. In addition, the applicability of SEP measurements is limited by patients' physical status and cooperation. The clinical significance of posterior tibial nerve SEP testing in poststroke hemiparetic patients is therefore debatable. PMID- 9339165 TI - Reduction of the fatigue-induced force decline in human skeletal muscle by optimized stimulation trains. AB - OBJECTIVE: To identify the stimulation pattern that optimizes the force-time integral produced during isometric contractions of fatigued human skeletal muscle. DESIGN: Twelve healthy subjects with no history of lower extremity orthopedic problems voluntarily participated. RESULTS: The primary findings were that (1) the optimized trains showed augmentation only from fatigued muscles and (2) a simple stimulation pattern, containing one brief (5msec) initial interpulse interval, produced the greatest force-time integrals and rates of rise of force. With muscle fatigue, the rate of rise of force of the constant-frequency train slowed, whereas the rate of rise of force of the optimized trains remained unchanged. This difference in the rate of rise of force may explain why the optimized trains, which take advantage of the catchlike property of skeletal muscle, are able to augment forces from fatigued muscles when compared with the constant-frequency train. CONCLUSIONS: These results may have important clinical implications when using brief trains of electric stimulation to aid patients in performing functional movements and contribute to our understanding of the relationship between the activation pattern of a muscle and the force output produced. PMID- 9339166 TI - Performance of selected lightweight wheelchairs on ANSI/RESNA tests. American National Standards Institute-Rehabilitation Engineering and Assistive Technology Society of North America. AB - OBJECTIVE: This study provides data for clinicians and wheelchair users to compare the durability, stability, and cost effectiveness of three different lightweight wheelchair models: the Everest & Jennings EZ Lite, the Invacare Rolls 2000, and the Quickie Designs Breezy. A second objective was to compare the results from this study to those published for ultralight and institutional depot wheelchairs. DESIGN: Randomized standards testing of three wheelchair models from each manufacturer (nine wheelchairs total). RESULTS: There were no significant differences (p > .05) in fatigue life, life-cycle cost, or static stability between the three models of lightweight wheelchairs (ie, EZ Lite, Rolls 2000, or Breezy). There were, however, significant differences (p < .05) in fatigue life among the lightweight wheelchairs of this study and the published results for ultralight rehabilitation wheelchairs and for depot wheelchairs. The lightweight wheelchairs had an average fatigue life greater than the depot wheelchairs but less than the rehabilitation wheelchairs. A depot-type wheelchair was defined as a manual wheelchair designed for hospital or institutional use. At lightweight wheelchair was defined as a manual wheelchair with minimal adjustments designed for individual or institutional use. An ultralight rehabilitation wheelchair was defined as a manual wheelchair designed for an individual's use as a long-term mobility aid. CONCLUSION: The three models of lightweight wheelchairs tested are substantially similar and their fatigue lives are significantly (p < .05) lower than rehabilitation wheelchairs. Ultralight rehabilitation wheelchairs are the most cost effective over the life of the wheelchair, costing 3.4 times less (dollars per life cycle) than depot wheelchairs, and 2.3 times less (dollars per life cycle) than the lightweight wheelchairs tested in this study. PMID- 9339167 TI - Single-subject research in rehabilitation: a review of studies using AB, withdrawal, multiple baseline, and alternating treatments designs. AB - OBJECTIVE: To review the "methodologic rules" for using single-subject research designs (SSRDs) and to review the use of SSRDs in rehabilitation research of the past decade. DATA SOURCES: CINAHL and MEDLINE searches using "single subject" and "single system" as key words for the period 1985-1995 yielded 61 articles related to rehabilitation. STUDY SELECTION: Studies were selected for review if they described one of four commonly used SSRDs, specifically AB, withdrawal, multiple baseline, or alternating treatments. DATA EXTRACTION: Studies cited were identified by consensus and either exemplify adherence to the experimental rules of SSRDs or illustrate errors that result in threats to the validity of stated findings. DATA SYNTHESIS: All four types of SSRDs have been reported in rehabilitation studies, sometimes incorrectly. CONCLUSIONS: SSRDs, with their client-centered focus, are ideally suited for researching human behavior in the rehabilitation practice environment. Although numerous sources clearly identify the methodologic requirements for single-subject experiments, several studies violate the basic rules, threatening the validity of the results of these studies. Other properly applied SSRDs illustrate the strengths of this approach, which can produce empirical support for rehabilitation interventions. PMID- 9339168 TI - Posture effects on grip strength. AB - OBJECTIVE: To examine whether grip strengths were different when measured in supine and sitting positions. DESIGN: Comparison, convenience sample. SETTING: Community. PARTICIPANTS: Seventy-four healthy adult participants with no history of psychiatric or neurological dysfunction, or upper extremity orthopedic dysfunction after the age of 18. INTERVENTIONS: Participants performed grips with each hand while sitting and standing. Shoulder was adducted and extended, with the elbow flexed, and wrist and forearm in neutral. MAIN OUTCOME MEASURE: The mean of the three trials with each hand in each posture. RESULTS: Men were stronger (49kg) than women (29kg; p < .001). Right hands were stronger (41kg) than left (39kg; p < .001). However, grip strengths while sitting were equivalent to those tested while supine (p > .59). CONCLUSIONS: Using identical upper extremity positions, grip strength is equivalent when tested in the supine and sitting positions. Thus, when determining grip strength, grips measured while the subject is supine can be compared with norms collected while the subject is sitting, provided the upper extremity position is invariant. PMID- 9339169 TI - "Phantom" carpal tunnel syndrome. AB - Phantom sensation is ubiquitous among persons who have had amputation; however, if it develops into phantom pain, a thorough clinical investigation must ensue. We illustrate this with the case of a 49-year-old woman, 14 years after traumatic amputation of her left 2nd through 5th fingers, and 10 years after traumatic left transfemoral amputation. She had had phantom sensation in her absent fingers for years and developed progressive pain in her phantom fingers 3 months before presentation. Nerve conduction study revealed a high-normal distal motor latency of the left median nerve and a positive Bactrian test (sensitivity 87%). She was diagnosed with "phantom" carpal tunnel syndrome and treated with a resting wrist splint, decreased weight bearing on the left upper limb, and two corticosteroid carpal tunnel injections with marked improvement. Clinicians should recognize that phantom pain may be referred from a more proximal region and may be amenable to conservative management. PMID- 9339170 TI - Interrater reliability of the Tinetti Balance Scores in novice and experienced physical therapy clinicians. AB - OBJECTIVE: To examine interrater agreement of scores by physical therapy novices and experienced clinicians on videotaped and live performances of the balance portion of Tinetti's Performance Oriented Mobility Assessment (BPOMA). DESIGN: A reliability design was used to assess the interrater agreement and consistency of the BPOMA scores in an elderly population. SETTING: General community hospital and skilled nursing facility. PATIENTS: Twenty-six residents of a skilled nursing home, ranging in age from 66 to 99 yrs (mean = 80.4, SD = 6.8), participated in Phase 1. Twenty-four hospital inpatients and five residents of a skilled nursing home, ranging in age from 60 to 92 yrs (mean = 74.7, SD = 7.9), participated in Phase 2. RATERS: Three student physical therapists scored the patients in Phase 1. One student was designated the administrating rater (AR). The AR instructed, guarded, and scored the subjects. The other two students were the observing raters (ORs), whose role was to observe and score the subject's performances. Nine physical therapy clinicians, ranging from 0 to 6 years of experience, rated subjects in Phase 2. MAIN OUTCOME MEASURES: Consistency and agreement of BPOMA scores were compared between clinicians with varying levels of experience. In Phase I, BPOMA was scored on-site by three student physical therapists. In Phase 2, videotaped performances were scored by five physical therapists, one physical therapist assistant, and three student physical therapists. RESULTS: Phase 1 demonstrated fair to excellent kappa coefficients (.40-1.00) in all maneuvers across all raters. The ORs had higher agreement compared with the AR, ranging from good to excellent (.75-1.00). Phase 2 demonstrated fair to good kappa coefficients (.40-.75) in 5 of 8 maneuvers across all nine raters. When comparing proportion of observed agreement to evaluate the years of experience on rater agreement, there was no significant difference between clinician groups. CONCLUSIONS: Fair to good reliability of BPOMA scores occurred across many rates of varied experience with a small amount of training. PMID- 9339171 TI - Modified axillary crutches for an adolescent with bilateral congenital transverse deficiencies of the radius and ulna and no hands. AB - This report describes the construction of modified axillary crutches for a patient who had bilateral congenital transverse incomplete deficiencies of the radius and ulna and no hands. Before physical therapy, the patient underwent a left pangeniculate percutaneous epiphysiodesis of the distal femoral and proximal tibial physis, causing him to be temporarily non-weight-bearing on the left lower extremity. The patient lacked both hands; therefore, he was unable to use crutches. Traditional axillary crutches were modified with permanent bilateral casts at the axilla/arm region to facilitate use by the patient. Using the modified axillary crutches, he ambulated independently and climbed and descended stairs with contact guard assist, while maintaining the non-weight-bearing status of the left lower extremity. PMID- 9339172 TI - The Navajo Health and Nutrition Survey: research that can make a difference. PMID- 9339173 TI - Rationale, design and methodology for the Navajo Health and Nutrition Survey. AB - As recently as 1990, there was no reservation-wide, population-based health status information about Navajo Indians. To remedy this shortcoming, the Navajo Health and Nutrition Survey was conducted from 1991 to 1992 to assess the health and nutritional status of Navajo Reservation residents using a population-based sample. Using a three-stage design, a representative sample of reservation households was selected for inclusion. All members of selected households 12 y of age and older were invited to participate. A total of 985 people in 459 households participated in the study. Survey protocols were modeled on those of previous national surveys and included a standard blood chemistry profile, complete blood count, oral glucose tolerance test, blood pressure, anthropometric measurements, a single 24-h dietary recall and a questionnaire on health behaviors. The findings from this survey, reported in the accompanying papers, inform efforts to prevent and control chronic disease among the Navajo. Lessons learned from this survey may be of interest to those conducting similar surveys in other American Indian and Alaska Native populations. PMID- 9339174 TI - Intake of nutrients and food sources of nutrients among the Navajo: findings from the Navajo Health and Nutrition Survey. AB - Diet has been implicated in the etiology of chronic diseases in many populations, including the Navajo and other American Indian tribes. This report describes the current nutrient intake of the Navajo and identifies the primary food sources of key nutrients. In the Navajo Health and Nutrition Survey, interviewers obtained single 24-h diet recalls from 946 nonpregnant participants age 12-91 between October 1991 and December 1992. Among various sex and age groups, total fat contributed 33-35% of energy and saturated fat contributed 10-11% of energy in the diets. Median fiber intake was 11-14 g/d. Median intakes of vitamin A, vitamin E, vitamin B-6, folate, calcium and magnesium were below sex- and age specific recommended dietary allowances (RDA) for men and women of all age groups. Intake of vitamin C was below the RDA for men and women age 20 and older. Median iron intake was below the RDA for women under age 60. Fruits and vegetables were each consumed less than once per day per person, as were dairy products. Fry bread and Navajo tortillas, home-fried potatoes, mutton, bacon and sausage, soft drinks, coffee and tea provided 41% of the energy and 15-46% of the macronutrients consumed. Recommendations to increase the intake of essential micronutrients in the Navajo diet are presented. PMID- 9339175 TI - Weight, body image, and weight control practices of Navajo Indians: findings from the Navajo Health and Nutrition Survey. AB - Historically, the Navajo exhibited a low prevalence of overweight, but a number of small studies over the past few decades indicate that the prevalence is increasing. In the population-based Navajo Health and Nutrition Survey conducted in 1991-92, overweight was defined as a body mass index (BMI, kg/m2) at or above the 85th percentile (BMI > 27.8 for men, > 27.3 for women) of the Second National Health and Nutrition Examination Survey. One third of men age 20 and 39 and one half of men age 40 and 59, but fewer than 10% of men age 60 and older were overweight. Two thirds or more of women in all age groups were overweight. Nineteen percent of the participants underestimated their weight status (underweight, appropriate, overweight) relative to their BMI category and 17% overestimated their weight status. Women overestimated their weight status more often than men (P < 0.05), and participants age 20-39 overestimated their weight status more often than older participants (P < 0.001). Men and women age 60 and older preferred heavier body shape models as ideals of health more often than younger participants (P < 0.001). Nearly half of the participants, regardless of their weight status, reported that they were trying to lose weight; most reported using diet and exercise. Because overweight is an important risk factor for many chronic diseases, including diabetes mellitus, cardiovascular disease and cancer, primary prevention of overweight and weight management for adults are recommended to prevent an increase in the burden of chronic disease among the Navajo. PMID- 9339176 TI - Risk factors for coronary heart disease among Navajo Indians: findings from the Navajo Health and Nutrition Survey. AB - Coronary heart disease was uncommon among the Navajo in the past, but appears to have increased substantially over the last few decades. The 1991-1992 Navajo Health and Nutrition Survey, which included interviews and examinations of 303 men and 485 women between the ages of 20 and 91 y, is the first population-based examination of coronary heart disease risk factors in this tribe. Coronary heart disease risk characteristics were common, particularly overweight (men, 35%; women, 62%), hypertension (men, 23%; women, 14%) and diabetes mellitus (men, 17%; women, 25%). Among 20- to 39-y-olds, a large proportion of men reported that they currently smoked cigarettes (23%); use of chewing tobacco or snuff was also prevalent among these 20- to 39-y-old men (37%) and women (31%). Although serum concentrations of total cholesterol were fairly comparable to those seen in the general U.S. population, fasting serum triglyceride concentrations were high (median: men, 132 mg/dL; women, 137 mg/dL), and concentrations of HDL cholesterol were low, particularly among women (median: men, 42 mg/dL; women, 44 mg/dL). Body mass index was associated with levels of most risk factors, and, independently of the level of overweight, a truncal pattern of body fat was related to adverse lipid levels among men. A large proportion of men (20%) and women (30%) reported not having participated in physical activity during the preceding month. Lessons learned from past intervention activities among the Navajo, particularly those for diabetes, may be useful in managing these risk factors to reduce the future burden of coronary heart disease. PMID- 9339177 TI - Diabetes mellitus among Navajo Indians: findings from the Navajo Health and Nutrition Survey. AB - Noninsulin-dependent diabetes mellitus is a major health problem among most American Indian tribes. This is the first population-based reservation-wide study of the Navajo that has used oral glucose tolerance testing to determine diabetes status. Employing WHO criteria, we found an age-standardized prevalence of diabetes mellitus (DM) of 22.9% among persons aged 20 y and older. This prevalence is 40% higher than any previous age-standardized estimate for the Navajo and four times higher than the age-standardized U.S. estimate. More than 40% of Navajo aged 45 y and older had DM. About one third of those with DM were unaware of it, with men more likely to be unaware than women. Among persons with a medical history of DM, almost 40% had fasting plasma glucose values > or = 200 mg/dL. Persons with DM were heavier, more sedentary and more likely to have a family history of DM than were persons without DM. Persons with DM had more hypertension, lower HDL levels and higher triglyceride levels than their counterparts without DM. Insulin usage was infrequent among persons with a history of DM, and about one third of women with such a history used no medical therapy to control their diabetes. Although important measures to combat diabetes have already been undertaken by the Navajo, additional efforts are required to slow the progression of this disease and prevent its sequelae. PMID- 9339178 TI - Prevalence of hypertension among Navajo Indians: findings from the Navajo Health and Nutrition Survey. AB - Hypertension and other chronic diseases are becoming increasingly important health problems for many Native American people, including the Navajo. A community-based survey that included three standardized measurements of blood pressures, was conducted during 1991-92 on the Navajo Reservation. Among the 780 adults examined, the overall age-standardized prevalence of hypertension, defined as an elevated systolic (> or = 140 mm Hg) or diastolic (> or = 90 mm Hg) blood pressure, or possession of prescription antihypertensive medications, was 19% (24% among men and 15% among women). The prevalence of hypertension increased with age and relative weight, and among men, was associated with diabetes mellitus. Among women, hypertension was associated with a central distribution of body fat, cigarette smoking, self-reported diabetes mellitus and impaired glucose tolerance. Although only 50% of the persons found to have elevated blood pressure at the examination reported they had been previously told that they had hypertension, persons who had been previously diagnosed with hypertension had a slightly higher rate (approximately 60%) of blood pressure control than that seen in the general U.S. population. On the basis of these results, the prevalence of hypertension among the Navajo appears to have substantially increased since the 1930s. Improved prevention and management of hypertension, especially for overweight and diabetic individuals, may reduce morbidity and mortality from cardiovascular and renal disease. PMID- 9339179 TI - Obesity, levels of lipids and glucose, and smoking among Navajo adolescents. AB - Although there is a high prevalence of overweight among Navajo children and adolescents, other risk factors for chronic disease in this population have received little attention. We therefore examined the distribution and interrelationships of overweight, cigarette smoking, blood pressure and plasma levels of lipids and glucose among 160 Navajo 12- to 19-y-olds. In agreement with previous reports, participants were approximately 2 kg/m2 heavier than adolescents in the general U.S. population, and the prevalence of overweight (> 85th percentile) was 35-40%. Levels of total cholesterol and blood pressure were similar to those in the general U.S. population, but Navajo adolescents had a 5 10 mg/dL lower median level of HDL cholesterol, and a 30 mg/dL higher median triglyceride level. Eight percent of the adolescents examined had either impaired glucose tolerance or diabetes mellitus as assessed through an oral glucose tolerance test (n = 10) or self-report (n = 1). Relative weight (kg/m2) was associated with adverse levels of lipids, lipoproteins and glucose, with overweight adolescents having a fivefold greater risk for elevated triglyceride levels than other adolescents. Tobacco use was fairly prevalent among boys (24% cigarettes, 23% smokeless tobacco), but not girls (9% cigarettes, 3% smokeless tobacco). Because of its associations with other risk factors and with various chronic diseases in later life, it may be beneficial to focus on the primary prevention of obesity among Navajo children and adolescents. PMID- 9339180 TI - The health of Navajo women: findings from the Navajo Health and Nutrition Survey, 1991-1992. AB - Cancer-screening behaviors, reproductive history, risk behaviors during pregnancy and chronic disease risk factors were examined in a representative sample of 566 Navajo women residing on the Navajo Reservation in 1991-1992. Among all women 15 y and older, 59% were overweight, 4% were current smokers, 10% currently used smokeless tobacco and 12% were anemic. Seventy-one percent of Navajo women aged 18 and older reported ever having had a Pap smear, but only 35% of women aged 50 and over reported ever having had a mammogram. Among parous women, the prevalence of having received no prenatal care for any pregnancy declined from 60% among women 60 and older to 13% among women 20-29 y of age, and the prevalence of ever having had a child born at home declined from 82 to 2%. These data suggest marked secular improvement in these pregnancy-related risk behaviors. However, data on cancer-screening behaviors indicate opportunities to improve health of Navajo women by increasing their use of mammography and Pap smear screening services. PMID- 9339181 TI - [Value of the combination of oral ondansetron with methylprednisolone as soon as the first cure in mild emetogenic chemotherapy. Groupe francais d'etude de l'ondansetron]. AB - This multicentre randomized single-blind parallel group study compared the efficacy of oral ondansetron plus methylprednisolone (OND+MPS) with conventional antiemetic strategies (TH) over 4 consecutive courses in moderately emetogenic chemotherapy. This study was conducted in naive patients receiving a minimum of 3 cytotoxics including adriamycin (> or = 35 mg/m2) and cyclophosphamide (> or = 500 mg/m2) plus an other alkylating agent. Of the 364 patients included in the study, 70% had a breast cancer and 30% a lymphoma. Patients were divided into two groups. On day 1, one group of patients received OND (8 mg, po) 2 hours before chemotherapy, followed by a slow intravenous injection of MPS (120 mg) 30 minutes before chemotherapy. Eight hours after the start of chemotherapy, patients received OND (8 mg, po) and MPS (16 mg, po). On days 2-4, patients received OND (8 mg, po) and MPS (16 mg, po) twice daily. The second group of patients received conventional antiemetic treatment (benzamide plus corticosteroids with or without benzodiazepins). The primary efficacy parameter was defined as complete control of emesis (0 emetic episodes) over 4 consecutive courses of chemotherapy. In the OND+MPS group, 63% of patients experienced complete control of emesis versus 33% in the TH group (p < 0.001). The secondary parameters (percentage of days with no emetic episodes, control of emetic episodes, grade of nausea at each course, patient preference and quality of life evaluation) were always significantly better in the OND+MPS treated group. The percentage of days without any emetic episode over the 4 courses of chemotherapy was 91% in the OND+MPS group and 75% in the TH group (p < 0.001). Ninety-two percent of patients from OND+MPS group preferred to continue their treatment versus 76% in the TH group (p < 0.001). Concerning the quality of life assessed by FLIC and FLIE questionnaires, the analysis showed a significant difference at the end of the treatment in favor of OND+MPS (p = 0.037 and 0.0075 respectively). This study showed the interest in using the combination OND+MPS right from the first course of moderately emetogenic chemotherapy. PMID- 9339183 TI - [Campaign for cervical cancer screening with vaginal smears in women aged 50-69 years in Isere (France). Results of the first round (January 1991-June 1993)]. AB - The department of Isere, which is involved since 1990 in a breast cancer screening campaign concerning women aged 50 to 69, has managed to associate a cervical and colorectal cancers screening program. The target sample size is 98,000 individuals. Women are asked to refer their general practitioner or gynaecologist for cervicovaginal smears. Each woman is invited at a screening interval of 30 months. The results of the first invitation (November 1990 December 1992) are reported. Thus 29,570 women did referred, so that the screening uptake is 30% and 20,083 women (68%) had Pap smears inside the screening program. 1.1% of the smears were unsatisfactory and 1.2% of the tests showed abnormalities. Ninety-six percent of the women who had been referred for further examinations have been followed up. Thirty-eight women (representing 17% of smears with abnormalities) had surgery (conisation, hysterectomy, Wertheim). Among them, 5 cases of invasive cervical carcinoma and 25 in situ carcinoma were detected. The detected cancer prevalence per 1,000 women screened is 1.5/1000. An organised screening program for cervical cancer in association with breast cancer screening, seems to be an effective way of increasing smears realisation in women aged 50 to 69, and of involving general practitioners in cervical cancer screening. PMID- 9339182 TI - [Inefficacy of exchange-transfusion in case of a methotrexate poisoning]. AB - We report on a case of methotrexate (MTX) intoxication occurring in a 19-year-old man treated for a leukemia. Exchange-transfusion (ET) was performed in attempt to remove the MTX from the body. This exchange-transfusion was unable to decrease the MTX plasma concentration. This inefficacy of ET in MTX intoxication is in contradiction with previously reported recommendations. However, this result is easily explained by MTX pharmacokinetics parameters. PMID- 9339184 TI - [Unusual development of nonseminomatous germ cell tumors of the testis (NSGCT)]. AB - The authors describe a clinical case of metastatic testicular non-seminomatous germinal tumor with several recurrences treated by the association of chemotherapy and surgery for remaining lesions. Based on this case a review of the literature is done to illustrate the evolution of treatments, to highlight the evolutive potential of mature teratoma, the delayed recurrences. PMID- 9339186 TI - [Atypical mesoblastic nephroma with metastases right away?]. AB - The diagnosis of a metastatic kidney tumor arising in a 2-month infant is discussed between atypical mesoblastic nephroma and clear cell sarcoma. The precocity of distant metastases, their location in bone marrow, liver and thoracic soft tissues, and their association with myelofibrosis set up an original clinical presentation which seems to have never been described elsewhere. Treatment strategy with surgery of the primary followed by a polychemotherapy combining vincristin-etoposide-ifosfamide and the short term follow-up are reported. PMID- 9339187 TI - [Cancer and primary humoral immunodeficiency]. AB - The occurrence of cancers in patients with primary humoral immunodeficiency syndrome (X-linked agammaglobulinemia, common variable immunodeficiency, IgA deficiency) is more than mere coincidence. An increased risk of non Hodgkin's lymphoma and gastric adenocarcinoma, particularly for patients with common variable immunodeficiency, is well established. A literature review is presented on this subject with analysis of case reports, prospective and retrospective studies, and data from the Minnesota Immunodeficiency Cancer Registry. PMID- 9339185 TI - [Second primary cancers after small-cell lung cancer]. AB - The records of 1,371 patients with small cell carcinoma of the lung (SCLC) treated between 1983 and 1994, were reviewed for the occurrence of second primary malignancies (SPM). One was excluded for analysis because of insufficient data. Eight synchronous SPM (SSPM) and 8 metachronous SPM (MSPM) were identified, SSPM included non-small cell lung cancer in 6 patients, 1 head and neck cancer and 1 oesophageal cancer. Median survival after the diagnosis of SSPM was 6 months. The MSPM were detected between 1 and 6 years after the diagnosis of SCLC. MSPM included lung cancer (3 patients), gastrointestinal malignancies (2 patients), 1 hematologic malignancy, 1 prostatic cancer and 1 head and neck cancer. The median survival time after the diagnosis of MSPM was 4 months. Occurrence of SPM is a singular pattern of patients with SCLC. Tobacco consumption, genetic factors and carcinogenic effects of multimodality treatment are supposed mechanisms to explain SPM. PMID- 9339188 TI - [French screening program of breast cancer. Results from 5 districts (1989 1994)]. AB - Randomized clinical trials have demonstrated that screening by mammography can reduce the breast cancer mortality rate by 30 to 50% in women aged 50 to 69 years. In France, pilot screening programmes were begun in 1989, and a national programme is being implemented since 1994. After 8 years of experience, the present study aims to assess the effectiveness of the French decentralized model of screening. This study presents data obtained by the National Steering Committee from the reports of five district programmes. Efficacy indicators from 1989 to 1994 are presented and compared with European targets. Data are presented by screening round. The prevalent round concerns 260,226 women screened, the subsequent incident rounds concern 119,761 women. Attendance rates at 5 years range from 24.2% to 50.9%. The mean recall rate was 7.8% in the first and 4.5% in the second round 5.4 and 2.7 cancers were detected per 1,000 women screened during the first and second round, respectively. and 30% of invasive cancers were of 10 mm or less and 70% had no nodal involvement. The interval cancer rate was 0.55% one year after screening and 1.05% two years after screening. Results from our decentralized screening system are variable yet satisfactory overall, namely thanks to the progressive implementation of quality assurance. However, attendance rates remain low, whereas actual mammographic coverage is 60% in France. A generalized screening programme can only be foreseen in France if its implementation is very gradual and if the quality of screening is high. The improvement of radiological quality remains the primary objective of the National Steering Committee however this implies training of all personnel. In the long run, organized screening should prevail over spontaneous screening. It is unlikely that we will achieve the target reduction in mortality rate in our country. PMID- 9339189 TI - [Protein kinase C and tumorigenic potential]. AB - Initially described as a unique entity, protein kinase C (PKC) is now represented by a family of 11 isoforms which differ in their structural and biochemical properties as well as in their tissular distribution, subcellular localization and substrate specificity. So far a lot of studies have attempted to approach the role of each of these PKC isoforms in the deregulation of growth signaling that leads to carcinogenesis. Among the various strategies developed, the surexpression of specific isoforms in different cellular models is the strategy which led to major advances as concern the PKC-cancer relationship. This review reports the main results obtained in this field especially in that of colorectal cancer. PMID- 9339190 TI - [Docetaxel symposium]. PMID- 9339191 TI - [Molecular typing of tumors: possibilities and problems]. PMID- 9339192 TI - [Cancerous Philotecte]. PMID- 9339193 TI - [Automation of the cytometric analysis of the DNA content of solid tumors]. AB - An automatic machine, dedicated to solid tumor DNA ploidy quantitation has been built in order to provide pathologists with a tool usable in clinical practice. Main efforts were focused on an automation of each step of the analysis and on an elimination of any subjective choice, while preserving the quality of measurement. As the software is independent of the machine architecture, it offers performances which increase in parallel with the rapid evolution of the computers. An illustration of the various functionalities of the automaton is proposed through the study of deparaffined breast cancer samples. PMID- 9339194 TI - [Flow cytometry in ORL cancers]. AB - The possibility to perform flow cytometry was examined in a series of 167 patients with primary untreated head and neck carcinoma referred to our Institution from February 1989 to January 1992. In all cases, flow cytometry was carried out on frozen tumour samples. The Cox model was used including age, tumour size, nodal status on clinical assessment, topography, treatment, malignancy grade, S phase fraction and ploidy as independent variables and overall survival as dependent variable. In this study, ploidy could be assessed in only 73% of cases and S phase fraction and G2M in 65% of the population studied. No correlation could be evidenced between ploidy or SPF with other clinical, pathologic characteristics or clinical outcome. We conclude that flow cytometry should remain a research tool until the method has proved to be relevant in clinical routine, and until the yield of the technique can be improved. PMID- 9339195 TI - [Bayesian estimation of pharmacokinetic parameters of etoposide]. AB - Studies of the relationships between the pharmacokinetics of a drug and its pharmacodynamics could significantly improve chemotherapy efficacy. However, despite their proven value, pharmacokinetic studies sometimes appear as cumbersome and difficult procedures. The bayesian approach associated with an optimal sampling time strategy (OST) allows the determination of the pharmacokinetic parameters of a drug with a smaller number of blood samples compared with that required by the classic maximum likelihood estimation (MLE). Therefore, the bayesian approach may lead to a less discomfort to the patients and less work for the medical staff. Such a method was developed to determine the individual pharmacokinetic parameters of etoposide (VP16). First, the statistical characteristics of the pharmacokinetic parameters were evaluated in 14 courses from 14 patients. Then, based on these results, a three-sample strategy was developed. Validation of this methodology was performed in 7 new patients and evaluated by computing bias and precision. The performance of the developed methodology shows that it could successfully be applied for the determination of VP16 pharmacokinetic parameters. PMID- 9339196 TI - [Mediastinal large B-cell lymphoma. A retrospective anatomic-clinical study of 26 cases]. AB - Mediastinal B-cell lymphomas (with or without sclerosis) have been recently recognized as an entity with particular clinical features. We report 26 patients with a mediastinal large B-cell lymphoma. They represent 5% of the patients with aggressive non-Hodgkin's lymphoma and 2% of all non-Hodgkin's lymphoma seen in our centre between 1962 and 1990. They include 19 females (73%) and 7 males (27%). The sex ratio was 2.7 and the median age was 44 years (range: 17-84 years). Compressive symptoms in relation with a bulky mediastinum were present in 21 cases (80%) and with B symptoms in 5 cases. All these patients received 2 to 4 cycles of chemotherapy with a CHOP-like protocol (epirubicin or doxorubicin, cyclophosphamide, vincristine and prednisone) followed in 24 cases by mediastinum irradiation (40 Gy). Two patients progressed during chemotherapy and did not receive radiotherapy. Nineteen patients had a consolidation chemotherapy according to the same protocol. Twenty-one patients achieved a complete remission after chemotherapy or radiotherapy and 5 failed. Two patients relapsed at 10 months and 9 years. Seventeen patients are alive and in first complete remission with a median follow-up of 102 months (range: 60-260 months). Using the Kaplan Meier method, the overall survival at 5 and 10 years was respectively 77 and 61% and the relapse-free survival was respectively 68 and 57%. These results confirm the previous findings concerning this distinct entity which is characterized by a predilection for young women, compressive symptoms, a slow response to treatment and a rather good prognosis. PMID- 9339197 TI - [A medico-economic evaluation of second line chemotherapy in metastatic breast cancer: comparison between docetaxel, paclitaxel, and vinorelbine]. AB - Despite health public problems arised by metastatic breast cancer, specific studies remain rare, and especially when concerning second line chemotherapy. Today, these studies seem essential to allow the clinician, facing the choice between different treatments, to make the best decision. Two recent treatments, docetaxel and paclitaxel, administered every 3 weeks, were compared to the referenced treatment: vinorelbine administered every week. The study aims at evaluating these 3 therapeutic options on second line treatment of metastatic breast cancer. For each of these 3 strategies, our end points were: (1) the duration of progression free survival; (2) the quality adjusted on progression free survival; (3) the cost including the intrinsic cost of chemotherapy as well as the cost of treatment-related and disease-related complications, due to toxicity. Savings obtained with the treatment by delaying relapse were subtracted from expenditure. We used a Markov model to describe the patient's evolution after the administration of the compared treatments. The evaluation concerns the period from the beginning of second line chemotherapy to death. Only direct medical costs were taken into account. Non medical costs and indirect costs were excluded. For each clinical state, resources utilisation was estimated by a retrospective multicentric analysis of 153 medical records of metastatic breast cancer, treated on second line. Resources valuation of hospital costs were based on a national survey on the cost of medical services per DRGs. Quality of life was estimated by a group of nurses in oncology, using the feeling thermometer and standard gamble technics. Incremental cost utility ratios were calculated. Docetaxel reduces the time spent in progression, decreases the number of complications due to progressive disease and thereby provides better quality of life. It provides a benefit of 57 disease- and discomfort-free days, compared to vinorelbine and 22 days compared to paclitaxel. Docetaxel may be thought of as a self-financing strategy as a result of savings in hospital admissions, providing net saving of French Francs (FF) 6,800 in 1993 prices, compared with expenditure associated with vinorelbine treatment and FF 700 compared with the equivalent figures for paclitaxel. A sensitivity analysis confirms the robustness of those results. PMID- 9339198 TI - [Prognostic value of early normalization of CA 125 during chemotherapy in stages III and IV ovarian tumors]. AB - The value of early CA 125 assays and analysis of its diminution kinetics during chemotherapy have been the subject of numerous studies. In contrast, routine utilization of CA 125 assays in clinical practice remains controversial or at best difficult to apply because the definitions and prognostic values associated with CA 125 assays vary greatly from one study to the next. This study was designed to determine whether serial CA 125 assays during induction chemotherapy for ovarian carcinoma, using simple evaluation criteria directly applicable in routine clinical practice such as early normalization (level < 35 UI/ml) are predictive of response to treatment or improved survival. This retrospective longitudinal analysis concerned a historical population of 140 patients with ovarian carcinoma stages III and IV treated at the Antoine-Lacassagne Cancer Center between 1978 and 1993. All the patients were treated by chemotherapy based on platinum salts every 21 days. Serum CA 125 assays were performed both before and after surgery and during each chemotherapy cycle. Eighty-four patients had a pre-operative CA 125 assay. No difference is observed in survival as a function of their preoperative CA 125 concentration (p = 0.4). Sixty-seven patients had a CA 125 assay the 6th week after initiation of chemotherapy, 62 the 9th week and 47 the 18th week. Normalization of CA 125 the 6th week (p = 0.0001), the 9th week (p = 0.0008) and the 18th week (p = 0.03) after the initiation of the chemotherapy cycle are correlated with survival. The median survival in our study is 42 months if the CA 125 is below 35 UI/ml the 6th week versus 13 months if the level of CA 125 remains more than 35 UI/ml. In all, 66 of the 105 FIGO stage III patients underwent second-look surgery. Normalization of CA 125 levels is correlated with the absence of any gross residual tumor at the second-look procedure, the 6th week of chemotherapy (p = 0.0019), the 9th week of chemotherapy (p = 0.0003) and the 18th week of chemotherapy (p = 0.0015). This correlation is not confirmed when the presence of histologic residual tumor in biopsy specimens obtained during second-look surgery is taken into consideration. Overall, 88% of patients whose CA 125 levels failed to normalize during evaluation at the second cycle of chemotherapy have residual tumor at second-look surgery. Outside of clinical trials, repeated early CA 125 assays to determine the chemosensitivity and the prognosis of patients with ovarian carcinoma are of little interest compared to a single CA 125 assay at the 6th week after initiation of chemotherapy. This approach seems to be a good compromise between the information sought and its practical use. However the interest of early modification of chemotherapy regimen after 2 cycles, if the level of CA 125 remains more than 35 UI/ml, will have to be showed. PMID- 9339199 TI - [Efficiency of high-dose FEC chemotherapy for the mobilization of hematopoietic stem cells into peripheral blood]. AB - The best regimen for mobilizing hematopoietic stem cells (HSC) into peripheral blood is not yet defined. The efficiency of FEC chemotherapy in the treatment of breast cancer is well established and the effects of FEC on HSC mobilization have been characterized. We tested the feasibility and the toxicity of a high-dose FEC regimen which may improve the mobilizing capacity of conventional FEC. Twenty patients with poor prognosis breast cancer received high-dose FEC and filgrastim 5 micrograms/kg. Three leukaphereses were performed on each patient for 3 consecutive days. Total numbers of CFU-GM and CD34+ cells were assessed, and a retrospective analysis was made. The numbers of CFU-GM/kg and CD34+ cells/kg collected (mean +/- standard error) were respectively 12.2 x 10(5) (+/- 2.4) and 14.6 x 10(6) (+/- 2.5). Extra-hematologic toxicity was negligible. Hematologic recovery after CTCb high-dose chemotherapy and HSC infusion was rapid. High-dose FEC is efficient for collecting HSC in peripheral blood. Extra-hematologic toxicity is weak and hematologic recovery after autograft is normal. Increased dosage of epirubicin and cyclophosphamide could allow a single leukapheresis collection of sufficient HSC from peripheral blood. PMID- 9339200 TI - [Li-Fraumeni syndrome]. AB - The Li-Fraumeni syndrome is an autosomal dominant syndrome representing a genetic predisposition to a wide spectrum of tumours including sarcomas, breast carcinomas, brain tumors and adrenocortical carcinomas. In most of the cases, tumours will develop in children and young adults. Germline mutations of the tumor suppressor gene p53 have been identified in approximately 50% of the families. In most of the cases, germline p53 mutations are missense mutations, located between exon 5 and exon 8, within the DNA-binding domain of p53. Since these mutations inactivate the transcriptional activity of the protein, they can easily be detected by analyzing in yeast the transcriptional competence of p53 cDNA derived from lymphocytes. The presence of a germline p53 mutations must be considered in: (1) families including two first degree relatives with cancers belonging to the Li-Fraumeni spectrum, one relative being affected before age 45; (2) children or young adults with a rare tumour of in the general population, belonging to the Li-Fraumeni spectrum, such as adrenocortical carcinoma; and (3) children or young adults under age 45 with multiple primary tumours of the Li Fraumeni spectrum. Identification of a germline p53 mutation in an affected subject allows to establish the diagnosis of the Li-Fraumeni syndrome on a molecular basis. PMID- 9339202 TI - [P53 at the atomic scale: the multiple faces of a crystal]. AB - The p53 protein is a transcription factor activated in response to DNA-damaging agents (such as chemical or physical carcinogens) and which plays multiple role in the control of proliferation and survival of cells exposed to genotoxic stress. Recent developments in the analysis of the crystal structure of p53 help us to understand the exact role of the various domains of the protein, as well as the impact of the mutations which are frequently found in cancers. In the future, this structural approach may significantly contribute to the interpretation of the pathological consequences of p53 mutations. PMID- 9339201 TI - [P53 mutations, asset or disadvantage for cancer chemotherapy]. AB - The "guardian of the genome" p53 is an essential modulator of the cellular response to cytotoxic agents. After introduction of DNA damages, the p53 protein prevents the cells to divide, either transiently by arresting their progression at the G1/S transition, or definitely by inducing apoptosis. In approximately half of the human tumors, mutations result in profound alterations of the p53 protein properties. In this work, the consequences of these alterations on the tumor cell sensitivity to chemotherapy are discussed. PMID- 9339203 TI - [Tumor suppressor p53 gene: a potential target for cancer therapy?]. PMID- 9339204 TI - Cancer of the pyriform sinus: trends towards conservative treatment. AB - Despite progress in diagnostic methods, radiotherapeutic and surgical techniques, and the development of new chemotherapeutic agents, the prognosis of pyriform sinus carcinomas has not improved in a significant way over the last 2 decades. Whatever the treatment approach, for all stages combined, the overall survival at 5 years remains modest and rarely exceeds 30%. Deaths from distant metastases, second cancers and intercurrent diseases represent 30-40% of cases. For this, any improvement in loco-regional control does not necessary translate into a gain in survival. Thus, for these patients it is judicious to consider not only the loco regional control but also the quality of life since larynx preservation is concerned. In this review article, the results of different treatment approaches are illustrated by the most representative series in the recent literature. Early stages (T1-2) are managed conservatively either by radical radiotherapy or conservative surgery, although the former is the most frequently utilized as it requires less stringent patient selection. For more advanced stages (T3-4), recent tendencies appear to lean toward the use of primary radical radiotherapy with or without chemotherapy, with surgery reserved for persistent or recurrent tumor. However, this strategy is still under study and the initial results should be confirmed. PMID- 9339206 TI - [Advantage of using tumor markers in colorectal and breast cancers. Guidelines of the American Society of Clinical Oncology (ASCO)]. PMID- 9339205 TI - [Ataxia-telangiectasia and cancer: an open question]. AB - Ataxia-telangiectasia is a rare recessive disorder which, among other clinical signs, is characterized by an extreme sensitivity to ionising radiation. Cells isolated from AT patients show radioresistant DNA synthesis and this has lead to the hypothesis that the product of the genetic determinant of AT may play a role in the detection, signalling or repair of double stranded DNA breaks. The gene to AT, called ATM has been recently cloned and characterized. It codes for a large RNA transcript of 13,000 bp of which a 3,500 aa protein is translated. The gene itself covers 150 kb, spread over 64 exons. The amino acid sequence has revealed the existence, at the carboxyterminal end of the protein, of a domain presenting homology to PI-3 kinase. This characteristic has allowed the description of a new family of nuclear protein, in yeast, drosophila an human, functionally involved in DNA damage signalling. It is interesting to note that a vast majority of mutations described in AT patients lead to the truncation of the protein and consequently to the elimination of the PI-3K domain, thus suggesting an important role in the normal function of the protein. An important question linked to AT mutation concerns the cancer risk associated to heterozygous mutations. It is well established that AT patients, homozygous for the mutation, present a 100-200 fold increased risk of cancer. Epidemiological studies have described a 3-5 fold increase risk of cancer (particular breast cancer in women) associated to the heterozygous mutation. Knowing that the incidence of the heterozygotes can be estimated to range 0.5 to 1% in the general population this question is of great importance in terms of public health. PMID- 9339207 TI - [Tyrosine protein kinase inhibitors in cancerology: the end of the beginning?]. PMID- 9339208 TI - Validation of a self-administered lead exposure questionnaire among suburban teenagers. AB - Teenagers represent a unique population in which to evaluate lead exposure. A self-administered questionnaire was developed to evaluate the current and historic lead exposures of teenagers. This work evaluates the exposure questionnaire for both its ability to predict lead exposure and the accuracy of the teenage respondents. Subjects received the survey at school and were instructed to get assistance from their parents in questionnaire completion. Environmental samples (dust, soil, and water) were collected from 30 suburban Boston homes to evaluate the questionnaire's predictiveness. To evaluate the accuracy of subjects' responses, independent information about housing was obtained. The questionnaire was effective in identifying predictors of dust and soil lead levels, but not for water lead levels. Fine dust lead loading (< 150 microns) varied significantly among the six housing age categories (pre-1940, 1940-1949, 1950-1959, 1960-1969, 1970-1979, and > 1979) and traffic levels. Fine dust lead concentrations varied significantly with decade of housing construction. Mean soil lead levels varied significantly among housing age categories, traffic levels, and exterior construction materials. For the important predictors, there was excellent agreement between the teenagers' self report and confirmatory information. For housing age categories, the observed agreement was 69%; for traffic level, the observed agreement was 88%. These results illustrate that questionnaires continue to be useful in evaluating home lead levels even in suburban homes and that teenagers are accurate respondents. PMID- 9339209 TI - The El Paso smelter 20 years later: residual impact on Mexican children. AB - Although there has been considerable concern regarding cross-border industrial contamination between Mexico and the United States, there are remarkably few data. One notable case study is the smelter in El Paso, Texas. In 1974 blood lead levels higher than 40 micrograms/dl were detected in 52% of children studied near the smelter, in the adjacent Mexican community of Anapra in Ciudad Juarez, Chihuahua. Lead smelting at this plant was halted in 1985, and as a result, lead levels in air decreased sharply; consequently, children's exposure to lead and other metals should have diminished accordingly. In order to assess the effect of removal of lead emissions from the area, three geographical locations in Anapra, varying in distance from the smelter source, were evaluated for lead, arsenic, and cadmium levels in soil and for lead in blood of children. It was found that lead levels in soil were inversely correlated with distance from the smelter. Arsenic and cadmium levels in soil were constant among the three sectors. However, at residential sites closer to the smelter, a higher percentage of children was found with blood lead levels exceeding the Centers for Disease Control's action level of 10.0 micrograms/dl. In the sector closest to the border 43% of children had blood lead levels greater than 10.0 micrograms/dl. Although blood lead levels in children living in Anapra have dropped approximately fourfold in 20 years, our results indicate a moderate continued risk of lead exposure. This study demonstrates the persistent impact that may result from cross-border contamination and raises provocative questions regarding appropriate action and the responsibility for financing such action. PMID- 9339210 TI - Blood lead in Uruguayan children and possible sources of exposure. AB - Blood samples and questionnaire background data were collected from 96 children (age 2-14 years) living in urban, suburban, or rural areas with varying traffic intensity and industrial lead pollution in Uruguay. Spot samples of tap water were collected from the homes of 44 children, and samples of top soil were taken from seven areas. Samples of air-borne dust were collected in central and suburban Montevideo. Blood lead concentrations (B-Pb) in children ranged between 47 and 191 (mean 96) micrograms/L and exceeded in 36% of the children 100 micrograms/L, the intervention level adopted by the United States Centers for Disease Control. Lead in tap water ranged from 0.2 to 230 (mean 15) micrograms/L and exceeded in 39% of the samples the maximum level recommended by WHO, 10 micrograms/L. Lead pipes were used in parts of the water supply systems. Lead in air varied between different locations from 0.15 to 1.7 micrograms/m3, highest in the very center of Montevideo. The median soil lead ranged from 6 to 2100 micrograms/g and was highest in industrially polluted areas. At multiple regression analysis, B-Pb was significantly associated only with age (P = 0.032) and traffic intensity at school (P = 0.045). No significant impact on B-Pb of lead in water or soil could be established. PMID- 9339211 TI - Effects of ultrafine and fine particles in urban air on peak expiratory flow among children with asthmatic symptoms. AB - It has been suggested that ultrafine particles in urban air may cause the health effects associated with thoracic particles (PM10). We therefore compared the effects of daily variations in particles of different sizes on peak expiratory flow (PEF) during a 57-day follow-up of 39 asthmatic children aged 7-12 years. The main source of particulate air pollution in the area was traffic. In addition to the measurements of PM10 and black smoke (BS) concentrations, an electric aerosol spectrometer was used to measure particle number concentrations in six size classes ranging from 0.01 to 10.0 microns. Daily variations in BS and particle number concentrations in size ranges between 0.032 and 0.32 micron and between 1.0 and 10.0 microns were highly intercorrelated (correlation coefficients about 0.9). Correlations with PM10 were somewhat lower (below 0.7). All these pollutants tended also to be associated with declines in morning PEF. However, the only statistically significant associations were observed with PM10 and BS. Different time lags of PM10 were also most consistently associated with declines in PEF. Therefore, in the present study on asthmatic children, the concentration of ultrafine particles was no more strongly associated with variations in PEF than PM10 or BS, as has earlier been suggested. PMID- 9339212 TI - Immune system alteration in the rat after indirect exposure to methyl mercury chloride or methyl mercury sulfide. AB - Methyl mercury is a well-recognized health hazard. It is an environmental contaminant that accumulates in the food chain. The primary source of mercury exposure for humans is through the consumption of contaminated fish. We studied the effects of indirect methyl mercury exposure on the immune system of Sprague Dawley rats. The effects of different forms of methyl mercury on immune system development were studied in Sprague-Dawley rats at 6 and 12 weeks of age. Rats were indirectly exposed to mercury during gestation and during nursing by exposing pregnant rats to either 5 or 500 micrograms/liter of methyl mercury chloride (CH3HgCl) or 5 micrograms/liter of methyl mercury sulfide [(CH3Hg)2S] in their drinking water. Total body, splenic, and thymic weights were measured, and NK cell cytolytic activity and lymphoproliferative response to T and B cell mitogens were evaluated in the offspring. At 6 weeks of age, total body and splenic weights were significantly increased in both high- and low-dose methyl mercury chloride-exposed groups. Rats exposed to methyl mercury sulfide had a significant increase in thymic weight at 6 weeks of age. At 12 weeks, the total body and organ weights were not different from controls. The lymphocyte proliferative response of splenocytes to PWM was enhanced at 6 weeks in both CH3HgCl exposed groups and not affected in the (CH3Hg)2S exposed group. NK cell activity was not affected in either group at 6 weeks of age. At age 12 weeks, NK cell activity was statistically significantly decreased by 56.6% in both CH3HgCl exposed groups and not affected in the (CH3Hg)2S-exposed rats. The lymphocyte proliferative response of splenocytes to the B cell mitogen pokeweed remained increased in the CH3HgCl groups. Indirect exposure of rats (during gestation and nursing) to different forms of methyl mercury reveals that chloride forms have prolonged predominantly enhancing effects on lymphoproliferative response of splenocytes, followed by significant depression of NK cell activity. PMID- 9339213 TI - Influence of stress on DDT-induced humoral immune responsiveness in mice. AB - The effects of different durations/intensities of stress on DDT-induced modulation of humoral immune response were evaluated in mice. DDT (20, 50, or 100 ppm x 4 weeks) per se did not influence the primary antibody response to sheep red blood cells (SRBC). However, when DDT-pretreated mice were exposed to single and multiple sessions of restraint stress (RS), the anti-SRBC antibody titers were lower than the control values, the most prominent effects being seen after 50 and 100 ppm DDT exposure in combination with a single intense stressor (24 hr RS) or repeated stress (1 hr RS x 5). In the splenic plaque forming cells (PFC) assay, similar potentiations of DDT-induced immune suppression were seen at 50 and 100 ppm exposure levels in combination with 24 hr RS or 1 hr RS (x5) procedures. In addition, the 20 ppm DDT exposure effect was also potentiated in combination with the multiple RS model. Further, other forms of stress viz. 3 hr cold restraint stress (CRS) or 6 hr RS, which per se did not influence the antibody titer or PFC response, suppressed humoral immune responses, when combined with 100 ppm DDT exposure. These results are discussed in light of the possible interactions between physical/emotional and environmental/xenobiotic stressors in the regulation of humoral immune response. PMID- 9339214 TI - Prenatal exposure to methylmercury during late gestation affects cerebral opiatergic system in rat offspring. AB - Pregnant female rats were orally administered a single dose (8 mg/kg) of methylmercury chloride (MMC) on Day 15 of gestation. The binding characteristics of opioid receptors were studied in the brain of developing rats at different stages of age. An increased density of opioid receptors was found in whole brain of MMC-exposed rats at 21 days (delta receptors) and 60 days (mu and delta receptors) of age, in comparison with matched controls. An enhanced response to morphine administration was detected in MMC-exposed rat offspring at Day 60 of postnatal life, which, however, was not apparently due to an impaired liver metabolization or renal excretion. Hence, it is reasonable to surmise a possible correlation between receptor up-regulation and increased response to pharmacological challenge. These data seem to indicate that neurochemical alterations produced in the rat developing organism by prenatal exposure to methylmercury involve the opiatergic system which undergoes a supersensitivity phenomenon. This effect, which is not detectable in the first postnatal period, shows a delayed onset, being detectable only at the adult stage. These findings seem to indicate that pre- and postnatal methylmercury exposure induces latent neurochemical and behavioral alterations which could last even after the clearance of the metal from the brain. PMID- 9339215 TI - In vitro interaction of alveolar macrophages and Aspergillus fumigatus. AB - In vitro interaction of alveolar macrophages (AM) from rats with conidia from Aspergillus fumigatus and Aspergillus candidus as well as inert control particles (amorphous silica) of similar diameter (about 3 microns), was studied. Experimental observations showed that both kinds of conidia were phagocytized significantly faster by AM than were the control particles due to a faster rate of attachment, but even more so due to a faster rate of ingestion. Quantitative nitroblue tetrazolium reduction by AM reflecting their oxidative metabolism and oxygen radical release was increased in response to both kinds of conidia by a factor of 2-3 during the process of phagocytosis, as well as 24 hr after the onset of phagocytosis, compared to corresponding conditions with inert particles and to resting macrophages. The mean pH in phagolysosomes with each of the conidia tended to be higher after 3 hr but was significantly lower after 24 hr than in the phagolysosomes with the control particles. After 3 hr there was a considerable percentage (around 8%) of phagolysosomes with pH > or = 6.5 and after 24 hr there was still a small percentage (0.7%) of such phagolysosomes for each of the conidia. Such a fraction was not observed for the control particles. Electron microscopic studies showed passages between phagolysosomes and AM surface with both kinds of conidia. The occurrence of such unsealed phagolysosomes might explain the percentage of phagolysosomes with high pH. Hence, Aspergillus conidia in unsealed macrophage vacuoles mediate an increased oxygen radical release from the macrophages, a process which in the long run might cause lung damage. PMID- 9339216 TI - Dietary consumption of Lathyrus sativus seeds induces behavioral changes in the rat. AB - Neurolathyrism is a degenerative disorder due to an excessive consumption of Lathyrus sativus (LS) seeds, which contain the neurotoxic amino acid beta-N oxalylamino-L-alanine. In this study, a population of Wistar rats was fed a diet with LS seeds up to 8 months. Two control groups were chosen, one receiving standard food and the other Cicer arietinum seeds (a nontoxic legume). At the end of the dietary period, the groups previously fed the seeds were switched to standard food for 1 month (wash-out). All animals were submitted to a neurological examination and observed in an open-field situation before, during the diet (at 4 and 8 months), and finally after wash-out. Neither LS-fed rats nor controls ever showed neurological deficits. By contrast, in an open-field the activity was significantly increased in the LS-eating rats at both the 4th and 8th month. The effect was indeed reversible, since it disappeared after the wash out. It is suggested that the enhanced open-field activity seen in the LS group might indicate a reversible excitable status. However, there is no evidence at present that the behavioral changes described represent a marker of neurodegeneration in this animal species. PMID- 9339217 TI - Pathways of lead exposure in urban children. AB - A linear structural equation modeling procedure was used to explore the mechanisms and pathways for lead intake among urban children and the relative contribution of various lead sources to lead-contaminated house dust. Dust lead levels were significantly associated with children's blood lead levels, both indirectly and directly via hand lead. Both soil and paint lead contributed to dust lead levels, but paint contributed significantly more lead to house dust than soil (P < 0.001). Black race and income level both directly affected children's blood lead levels. Finally, time spent outdoors was associated with children putting soil or dirt in their mouths which was, in turn, associated with children's blood lead levels. These data indicate that mouthing behaviors are an important mechanism of exposure among urban children with low-level elevations in blood lead and that lead-based paint is a more important contributor of lead to house dust than is lead-contaminated soil. PMID- 9339218 TI - Coal mine workers' pneumoconiosis (CWP): in vitro study of the release of organic compounds from coal mine dust in the presence of physiological fluids. AB - Solvents like dichloromethane generally are used to yield exhausted extraction amounts of the organic compounds in coals. Leaching of coal mine dust by dichloromethane yields extracts with comparable amounts of alkanes, aromatics, and phenolic compounds. Dominantly phenolic compounds are leached from coal mine dust by aqueous solutions saturated in lecithin because of their high water solubility. High concentrations of phenolic compounds can be extracted from coal mine dust generated from low-rank coals. Phenolic compounds leached by fluids adapted to physiological conditions correlate with high cytotoxicities of the dust from low-rank coals. Adaptation of leaching fluids to physiological conditions allows a more realistic estimation of experiments. Coal mine dust with varying coal content of different ranks can be seen as a parameter reinforcing the cytotoxic potential of coal mine dust. PMID- 9339219 TI - Effect of in vitro exposure to tributyltin on generation of oxygen metabolites by oyster hemocytes. AB - Mollusks depend chiefly on hemocyte-mediated cytotoxic mechanisms such as reactive oxygen species (ROS) to defend against pathogenic microorganisms. The effect of in vitro tributyltin chloride (TBT) exposure on ROS generation by oyster (Crassostrea virginica) blood phagocytes is quantified in this study. Luminol-augmented chemiluminescence (LCL) was used to measure ROS activity of resting and zymosan-stimulated cells after 1 or 20 hr TBT exposure. LCL is thought to measure primarily the activity of the myeloperoxidase/hydrogen peroxide/ halide antimicrobial pathway. Hemocytes in TBT-free medium (controls) produced low level LCL, which was markedly stimulated by the addition of zymosan particles. Both resting and zymosan-stimulated LCL values were significantly inhibited by > or = 80 ppb TBT after either 1 or 20 hr of exposure. Exposure to < or = 2 ppb TBT concentrations for 20 hr produced slightly enhanced LCL activity, suggesting a hormesis-like effect. Partial reversibility of TBT suppression of LCL took place when previously exposed cells were put in TBT-free medium. The TBT concentrations used in these studies were not cytolethal in vitro and were considerably less than oyster tissue levels recorded after chronic, sublethal in vitro exposures. The data suggest that the common aquatic contaminant TBT can interact rapidly with C. virginica hemocytes to produce a partially reversible immunotoxicological lesion. Xenobiotic-induced suppression of ROS production by hemocytes may increase host susceptibility to infectious diseases. PMID- 9339221 TI - Clamped in a straitjacket: the insertion of lead into gasoline. AB - In 1925, shortly after production of tetraethyl lead began, an outbreak of psychosis occurred in all three operating plants, and a number of workers died in acute mania. A temporary moratorium on the production of tetraethyl lead was imposed, and the Public Health Service convened a meeting of scientists, public health officials, and industrial representatives to investigate the safety of the product. This paper reviews the steps leading to the insertion of lead into gasoline, the response of industry to the tragic poisoning, the actions of the public health authorities to investigate the episode, and the forces that resulted in the restoration of the additive to gasoline. PMID- 9339220 TI - Effect of lead on bone and cartilage in sexually mature rats: a morphometric and histomorphometry study. AB - The effect of exposure to lead on the longitudinal development of the femur and of its cartilage growth plate was studied in rats. A group of forty-five 50-day old female Wistar rats was divided into a control group of 20 rats and an experimental group of 25 rats fed a diet supplemented with 17 mg of lead acetate per kilogram of feed for 50 days. On Day 50 all rats were killed and their right femurs were dissected. The femurs were cleaned of soft tissue and femoral lengths were measured with a Vernier caliper and thickness of growth cartilage (GPC-Th micron) by histomorphometry. Final body weights were significantly (P < 0.05) lower in the control group than in the rats given the lead-supplemented diet. Femur length did not differ between groups. Histomorphometry of the femur showed that the thickness of growth cartilage was higher (P < 0.05) in the control group. These findings suggested lead-induced inhibition of growth plate development. The growth plate may be one of the key target tissues accounting for the adverse effects of chronic lead exposure on skeletal development. PMID- 9339222 TI - Brain involvement in organophosphate poisoning. AB - Organophosphate poisonings cause substantial morbidity and mortality worldwide; however, the neurological effects have not been clearly established. We have studied cerebral perfusion to investigate neurotoxic effects. Clinical effects, plasma cholinesterase activity, and brain single photon emission computerization tomography (SPECT) data were investigated in 16 patients with organophosphate poisonings. The subjects were from an adult intensive care unit in a university hospital. Cholinesterase activity in plasma was determined upon admission and then every day in the morning. Brain SPECT studies were performed during the first week, at the end of therapy, and 3 months after discharge. Patients were classified into 3 groups using a modified Namba classification: latent poisoning (Group A); mild and moderate poisoning (Group B); or severe poisoning (Group C). None of the 6 patients in Group A showed any symptoms; 3 patients in Group B had muscarinic and nicotinic effects; 5 patients in Group C had muscarinic, nicotinic, and central nervous system symptoms. The average plasma cholinesterase for Groups A, B, and C were 54.16 +/- 9.10, 42.2 +/- 12.02, and 13 +/- 4.84 U/ml, respectively (normal range of plasma cholinesterase is 40-80 U/ml). Only 1 patient from Group A required treatment with oxime; 2 patients from Group B and all patients in Group C were given oxime, atropine sulfate, and mechanical ventilation. In the brain SPECT studies, the patients in Group A showed fewer perfusion defect areas than did Group B and C patients. All cases showed perfusion defects especially in the parietal lobe. In addition, perfusion improvement took more time for Group C than for the other groups. The intensive care unit stays of Group C were statistically longer than for Groups A and B. We concluded that brain SPECT is a highly sensitive diagnostic method, together with clinical symptoms and plasma cholinesterase activity, for monitoring the clinical prognosis of organophosphate poisonings. PMID- 9339223 TI - Postural sway frequency analysis in workers exposed to n-hexane, xylene, and toluene: assessment of subclinical cerebellar dysfunction. AB - To clarify the effects of organic solvents on the postural balance system, 29 male sandal, shoe, and leather factory workers exposed to n-hexane, xylene, and toluene (solvent workers) were examined by computerized static posturography with sway frequency analysis. Concentrations of metabolites of solvents in urine samples taken from the workers in the morning before work ranged from 0.41 to 3.06 (mean, 1.20) mg/g creatinine (Cn) for 2,5-hexanedione, from 0.10 to 0.43 (mean, 0.19) g/g Cn for methylhippuric acid, and from 0.05 to 2.53 (mean, 0.37) g/g Cn for hippuric acid; estimated concentrations of n-hexane in workplace air ranged from 13 to 100 (mean, 40) ppm. Control subjects were 22 healthy males without exposure to solvents. With eyes open, postural sway with a frequency of 2 4 Hz in solvent workers was significantly larger than that in controls in the anteroposterior direction. With eyes closed, sway with a frequency of 0-1 Hz was significantly larger in solvent workers in the mediolateral and anteroposterior directions. Results of multiple regression analysis showed that with eyes open the 1- to 2-Hz and 2- to 4-Hz sways were related positively to 2,5-hexanedione and inversely with methylhippuric acid. The pattern of changes suggests that the vestibulocerebellar and spinocerebellar afferent systems are asymptomatically affected by n-hexane; the effect of n-hexane on the vestibulocerebellar system is possibly inhibited by xylene. PMID- 9339225 TI - Motor vehicle exhaust and chronic respiratory symptoms in children living near freeways. AB - To examine whether motor vehicle exhaust from freeways has an effect on respiratory health of children, a cross-sectional study was conducted. Children attending schools situated less than 1000 m from major freeways in the Province of South Holland were asked to participate. The selected freeways carry between 80,000 and 150,000 vehicles per day. Separate counts for truck traffic indicated a range from 8000 to 17,500 trucks per day. At a total of 13 schools, 1498 children were asked to participate. From these children, 1068 usable questionnaires were obtained. Chronic respiratory symptoms reported in the questionnaire were analyzed with logistic regression. Distance from the freeway and (truck) traffic intensity were used as exposure variables. Cough, wheeze, runny nose, and doctor-diagnosed asthma were significantly more often reported for children living within 100 m from the freeway. Truck traffic intensity and the concentration of black smoke measured in schools were found to be significantly associated with chronic respiratory symptoms. These relationships were more pronounced in girls than in boys. PMID- 9339224 TI - Higher milk intake during pregnancy is associated with lower maternal and umbilical cord lead levels in postpartum women. AB - Lead exposure and its deleterious effects continue to be a problem in many countries. The lack of effective and safe treatments for low-level intoxication has promoted environmental interventions to control different sources of lead. In this study we evaluated the effect of milk consumption in 1849 mother-and-child pairs participating in the lead surveillance program in Mexico City. The mean lead levels were 11.2 micrograms/dL for maternal blood lead (MBL) and 10.8 micrograms/dL in umbilical cord. The correlation between blood lead and umbilical cord lead was r = 0.74. Forty-eight percent of the MBL exceeded 10 micrograms/dL and 9.5% exceeded 20 micrograms/dL. Maternal blood lead was positively related to the use of lead-glazed ceramic were and to traffic exposure and was inversely related to the consumption of milk and orange juice. Women who reported the consumption of more than 7 glasses of milk per week had a blood lead level of 8.7 micrograms/dL; in comparison, those women who reported a consumption of less than 7 glasses per week had a blood lead level of 11.1 micrograms/dL. Similar findings were observed for lead measured in umbilical cord. The association between lead levels and milk intake remained unchanged after taking in consideration other predictors of blood lead. This study suggests that a simple intervention could reduce lead burden among women and their newborns. PMID- 9339226 TI - Pesticide prioritization for a brain cancer case-control study. AB - The incidence of brain cancer is rising in the United States while the causes remain largely unknown. Epidemiologic studies indicate that individuals working in agriculture have an increased risk of brain cancer. The National Institute for Occupational Safety and Health is conducting a case-control study of incident brain cancer cases in Iowa, Michigan, Minnesota, and Wisconsin to evaluate the risk associated with several environmental exposures, in particular agricultural pesticides. Hundreds of different pesticides are used in agriculture and it is not feasible to evaluate the association between brain cancer and exposure to each of these chemicals; therefore, a strategy was developed to identify which pesticides would be targeted in the study. First lists of pesticides were created, documenting usage in each of the four states and the United States as a whole, by using data from reports prepared by the U.S. Department of Agriculture and Departments of Agriculture and land grant colleges within the four states. Then the following factors were considered in prioritizing pesticides for evaluation in the study: total volume of use prior to 1985, ranking of use in the four states and the United States as a whole by pesticide category, and toxicological evidence of carcinogenic, teratogenic, or mutagenic effects. Pesticide usage prior to 1985 was determined to allow for a minimum 10-year latency for the incident brain cancer cases diagnosed in 1995 or later. The selected pesticides include 56 herbicides, 49 insecticides, 12 fungicides, and 17 fumigants, accounting for over 99% of the total pounds of herbicides and insecticides and over 98% of the total pounds of fungicides and fumigants applied pre-1985. Prompt lists of the pesticides are sent to study participants a few days before the study questionnaire is administered to allow them time to recall past use of pesticides; the lists include the common chemical names, trade names, the crops that the pesticides are most commonly used on, and the years that the pesticides have been marketed. The methods used to select this subset of 134 pesticides document historical usage and may be useful in prioritizing pesticides for other research studies. PMID- 9339227 TI - Respiratory and irritant health effects of a population living in a petrochemical polluted area in Taiwan. AB - Reported herewith are the results from an ongoing study of outdoor air pollution and the health of persons living in the communities in close proximity to petrochemical industrial complexes. To determine if there is an excess of adverse health outcomes in the population exposed to petrochemical industrial emissions, a health survey was undertaken in 1996 in this area and in one reference area which has no local industrial emissions. The subjects were 436 adults (30-64 years of age) living in the Sanwei area (exposed area) and 488 in Taicei (reference area). For several indicators of respiratory health, including cough, wheezing, and chronic bronchitis, the prevalence rates were not significantly different between the study and the control populations. Acute irritative symptoms (eye irritation, nausea, throat irritation, and chemical odor perception) were significantly more common in the exposed area, particularly perception of chemical odors (84.6% vs 2.1%). It is concluded that exposure to petrochemical air emissions may be associated with increased rates of acute irritative symptoms. Future studies are needed to identify the potential role of petrochemical industrial emissions (particularly volatile organic compounds) in the genesis of acute irritative symptoms in a nearby petrochemical industrial area. PMID- 9339228 TI - Urban air pollution enhances the formation of urethane-induced lung tumors in mice. AB - This paper investigates the association between air pollution and lung neoplasia in an animal model. The experimental exposures were done in two locations with different air pollution profiles: a polluted area (downtown Sao Paulo) and a "clean" environment (Atibaia). Swiss mice were employed and urethane (3 g/kg) was used as carcinogenic substance. Two experiments were performed: Experiment I was designed to verify whether air pollution acts as initiator and/or promoter of lung cancer, using 300 mice; Experiment II employed 250 animals and aimed to verify if the effects of air pollution on the development of lung tumors was dose dependent. A significant effect of air pollution in augmenting lung carcinogenecity induced by urethane was observed. This effect was shown to be dose-dependent and reproducible on two different occasions. In addition, morphometric studies revealed that pollution may influence tumor phenotype. These results support the hypothesis that air pollution plays a significant role in the development of lung tumors. PMID- 9339229 TI - Census tract analysis of lead exposure in Rhode Island children. AB - There has been increasing interest in a targeted approach to the screening and prevention of lead exposure in children. Targeted screening requires an understanding of variation in lead exposure in individual children or by region. In order to better understand variation by region, we studied Rhode Island lead poisoning screening data, examining average lead exposure to children living in 136 Providence County census tracts (CTs). The study population included 17,956 children aged 59 months and under, who were screened between May 1, 1992, and April 30, 1993. We evaluated the relationship between the percentage of children with blood lead > or = 10 micrograms/dL (pe10) and sociodemographic and housing characteristics, derived from United States 1990 Census data, of these CTs. CT descriptors included population density, percentage of households receiving public assistance income, median per capita income, percentage of households female headed, percentage of houses owner occupied, percentage of houses built before 1950, percentage of houses vacant, percentage of population Black, percentage of recent immigrants, and intraurban mobility. On average, 109 children were screened in each census tract; mean screening rate was 44%. There was wide variation in average lead exposure among census tracts, with pe10 ranging from 3 to 60% of screened children (mean 27%). Individual census variables explained between 24 and 67% of the variance in pe10 among CTs. A multiple regression model including percentage screened, percentage of households receiving public assistance, percentage of houses built before 1950, In (percentage of houses vacant), and percentage of recent immigrants explained 83% of variance in pe10. The percentage of houses built before 1950, a variable which models the presence of lead paint in old houses, displayed the largest adjusted effect on pe10 over the range observed for that variable in RI CTs. The percentage of houses vacant was also a highly significant and robust predictor; we suggest that vacancy is an ecological marker for the deterioration of leadbased paint, with higher vacancy neighborhoods containing houses in poorer condition. In Rhode Island, census tracts with high vacancy rates also have high rates of recent immigration, making immigrant groups vulnerable to lead exposure. Small-areas analysis may be useful in directing resources to high risk areas, explaining the sociocultural forces which produce such exposure and analyzing the effects of housing policy over time in states with high screening penetration. PMID- 9339230 TI - Solubilization of lead from crystal dust in protein solution (pseudointerstitial fluid) and gastric juice. AB - Since several workers engaged in polishing and engraving crystal articles were found to have higher than average blood levels of lead (560 micrograms/liter, range 80-560 micrograms/liter), we investigated the hypothesis that crystal dust releases lead in the human body. To test the hypothesis, two types of crystal polishing dusts, having different lead contents, were mixed with human serum diluted 1:3 (pseudointerstitial fluid), gastric juice, and phosphate buffer at pH 9. After 14 days of contact, the diluted serum had extracted 0.620% of the lead in the crystal dust (particle size < 20 microns) containing 25.2% lead and 0.425% of that containing 19.9% lead. After 48 hr in gastric juice, 0.235 and 0.556% of the lead was extracted from crystal dusts (unsieved crystal dusts) containing 25.2 and 19.9% lead, respectively. After 28 days in alkaline solution, 0.358 and 0.304% of the lead was extracted respectively from the same two crystal dusts (unsieved crystal dusts). PMID- 9339232 TI - Esophageal perforation after pneumatic dilatation for achalasia: why? AB - A retrospective study was performed to asses risk factors in patients with esophageal achalasia undergoing pneumatic dilatation. Of 140 patients who underwent 159 dilatations, 7 sustained esophageal perforation (4.4%). They were matched with a group of 52 non perforated, dilated achalasia patients. History of prior pneumatic dilatation and small esophageal diameter were found to be risk factors by chi square and ANOVA. CONCLUSIONS: 1) Pneumatic dilatation for esophageal achalasia is a procedure with 4.4% risk of perforation and 0.6% mortality rate. 2) The risk of developing an esophageal perforation is increased by previous pneumatic dilatation and small esophageal diameter. Another risk factor such as a possible anatomical weakness of the esophageal wall (and the likelihood of it being evaluated by ultrasonography) at the site of perforation is suggested. PMID- 9339231 TI - The effect of air pollution on the integrity of chlorophyll, spectral reflectance response, and on concentrations of nickel, vanadium, and sulfur in the lichen Ramalina duriaei (De Not.) Bagl. AB - The following study was designed to determine the environmental impact of pollutants emitted by combustion of heavy fuel oil in Ashdod, in Southwestern Israel. For this purpose we measured concentrations of total S, V, and Ni in the local epiphytic fruticose lichen Ramalina duriaei, which grows in the peripheral region of the town, and compared these results with those obtained in thalli collected 100 km away, in the HaZorea Forest, northeastern Israel. We also transplanted thalli from the HaZorea Forest to the Ashdod region for a 10-month period. At the end of the experiment we measured the elemental content in all samples. In addition we measured chlorophyll degradation expressed as changes in the 435 nm/415 nm OD ratio, and changes in the spectral responses of the thalli. In several sites in the Ashdod region we found high concentrations of S, V, and Ni in transplanted thalli, which correlated with the NDVI values. These findings agree with other measurements of SO2 and V in the Ashdod area. We suggest that a high V/Ni ratio in lichens is a tracer for air pollution caused by the combustion of heavy fuel oil. PMID- 9339233 TI - Rat colon epithelium response to hypoxia is modified by intrinsic innervation blockade. AB - BACKGROUND/AIM: Short-circuit current (Isc) and transepithelial potential difference (PD) of rat distal colon decrease during acute hypoxia and overshoot on reoxygenation. It is not known whether tonic intrinsic nervous activity may influence these responses. METHODS: Preparations lacking the submucosal plexus (islet mucosa) and preparations retaining it (mucosa-submucosa) were mounted in Ussing chambers at 37 degrees C and gassed with 95% O2-5% CO2; Isc and PD were monitored. A 5-min hypoxia with 95% N2-5% CO2 was followed by reoxygenation. The procedure was repeated in the presence of the nervous blocking agent, tetrodotoxin (10(-6)M) in the serosal side of the chamber. RESULTS: In the isolated mucosa (n = 10) hypoxia reduced Isc by -55 +/- 5% and PD by -54 +/- 6% below baseline; reoxygenatory overshoots were, respectively, +60 +/- 17% and +/- 16%. Tetrodotoxin slightly and transiently reduced baseline Isc (-16 +/- 2%) and PD (-14 +/- 3%), with a small resistivity increase. It did not significatively modify the responses to responses to either hypoxia or reoxygenation. In mucosa submucosa preparations (n = 9) hypoxia reduced Isc (-54 +/- 8%) and PD (-61 +/- 4%). On reoxygenation Isc and PD were increased, respectively, +30 +/- 5% and +19 +/- 6% over baseline. Tetrodotoxin reduced baseline Isc (-59.6 +/- 5%) and PD (61.3 +/- 6%). It enhanced hypoxic Isc and PD decreases (-80 +/- 5%), but not the reoxygenatory overshoots. CONCLUSIONS: 1) Tetrodotoxin affects baseline Isc and PD more intensely in submucosal plexus innervated preparations than in the isolated mucosa. 2) The epithelial electrical response to acute hypoxia appears to be modulated by tonic neural activity. PMID- 9339234 TI - Thioctic acid protection against ethanol-induced gastric mucosal lesions involves sulfhydryl and prostaglandin participation. AB - Thioctic acid, a sulfhydryl agent, given orally macroscopically protected the gastric mucosa from 96% ethanol-induced lesions in a dose-and time dependent fashion. The inhibition of the lesions was 56.0 and 90.3% at doses of 25 and 50 mg/kg, respectively. The duration of its protective effect was approximately 120 minutes. Histopathologically, the oral administration of thioctic acid prevented necrotic mucosal lesions in the deeper part of the mucosa but did not protect the surface epithelial cells against ethanol challenge. Gastric motility measured by a balloon method, was dose-dependently inhibited by the oral administration of thioctic acid. Thioctic acid protection was suppressed by pretreatment with indomethacin (30 mg/kg), a cyclooxygenase inhibitor, and iodoacetamide (100 mg/kg), a sulfhydryl blocker. The gastric motility inhibited by oral thioctic acid was not reversed by indomethacin or iodoacetamide. These doses of indomethacin or iodomethamide were administered because previously they had been used to suppress endogenous prostaglandins, and nonprotein sulfhydryls of the gastric mucosa, respectively. There was an increase in the fluid volume retained in the gastric lumen for thioctic acid (50 mg/kg) at 30, 60, 90, and 120 minutes after administration. There was an increase in the mucus volume retained in the gastric lumen for thioctic acid (50, 25 mg/kg) at 120 minutes after administration. The lesion area in the rats treated with 70 microliters of vehicle and in the rats treated with 250 microliters of vehicle were significantly higher than in the rats treated with 450 microliters of vehicle. The present study suggests that thioctic acid administered orally, offered protection to the rat gastric mucosa against 96% ethanol-induced lesions. This protective effect appears to be dependent on prostaglandin-and sulfhydryl sensitive mechanisms, together with an increase in both the fluid volume and the mucus volume retained in the gastric lumen, and is not associated with the inhibition of gastric motor activity. PMID- 9339235 TI - Hemostatic and hemodynamic effects of vasopressin analogue DDAVP in patients with cirrhosis. AB - Desmopressin (DDAVP), a synthetic analogue of vasopressin, has been shown to improve the bleeding time in patients with cirrhosis. The duration of this effect and the hemodynamic changes associated with DDAVP have not been studied so far. To evaluate these issues, 14 cirrhotics with portal hypertension were studied in basal conditions and after DDAVP (0.3 uk/kg). In 8 patients, hemostatic tests were done at basal conditions and 1, 3, 6 and 24 hs after drug administration. In the remaining 6 patients, mean arterial pressure, cardiac output, portal and femoral blood flows were evaluated. Hemodynamic parameters were measured by Doppler ultrasound. DDVP caused a marked decrease in bleeding time at 1, 3, 6 and 24 hs (14 +/- 9 vs 8 +/- 3, 7 +/- 4, 6 +/- 4 and 8 +/- 4 min, respectively); the decrease was maximal and statistically significant at 6 hs (55 +/- 15%, p < 0.02) after DDAVP infusion. Bleeding time reduction was observed in every patient studied. In the hemodynamic study, DDAVP caused a mild but significant decrease in mean arterial pressure (12 +/- 8%, p < 0.05); no significant changes were observed in the rest of hemodynamic parameters studied. These findings show that DDAVP can be used to shorten the bleeding time for a period of at least 24 hs in patients with cirrhosis, without deleterious hemodynamic effects. This beneficial effect may be of potential relevance in the medical management of patients with chronic liver diseases. PMID- 9339237 TI - Biochemistry of the evolution of Triatoma infestans. XII. Biosynthesis and secretion of a very high density lipoprotein. AB - Biosynthetic processes related to the production of an insect hexamerin, very high density lipoprotein (VHDL), have been examined in the fat body of fifth instar nymph and adult Triatoma infestans. Fat bodies were incubated in vitro with [3H]leucine and the incubation media were precipitated using a specific antiserum. The SDS-polyacrylamide gel electrophoresis followed by blotting on nitrocellulose showed that both larval and adult fat body secreted the VHDL subunit. Moreover, the radiolabel recovered in this subunit is indicative of the de novo synthesis. When the incubation medium was subjected to density gradient ultracentrifugation, a radiolabeled fraction was found at density 1.27 g/ml, value identical to the hemolymph circulating VHDL, indicating that the secreted apoprotein is combined with lipids. The SDS-polyacrylamide gel electrophoresis and immunoblotting of this fraction corroborated the presence of the VHDL apoprotein. These results demonstrate that the fat body of T. infestans is able to synthesize the protein subunit which is associated to lipids as a lipoprotein particle that is released into the medium as VHDL. PMID- 9339236 TI - Esophageal motility disorders in Mexican patients with Duchenne's muscular dystrophy. AB - OBJECTIVE: Myopathies are entities that mainly involve striated muscle. In Duchenne's muscular dystrophy (DMD) there have been reported smooth muscle alterations in the pre-oral phase of swallowing, in gastric emptying, and pseudoobstruction. Nevertheless, esophageal motility alterations are not concluding. The objective of this work was to determine if there are motor esophageal alterations in this patients, and if this alterations are related to the clinical manifestations of disease. STUDY DESIGN: Nine consecutive patients with DMD (mean age 8, range 6-11 years; males) were evaluated, comparing clinical and manometric findings. RESULTS: Esophageal manometry alterations were found in all patients, mainly simultaneous non-peristaltic waves (60.86%) of diminished amplitude, in both striated and smooth muscle. Seventy seven percent presented with upper and lower gastrointestinal symptoms (dysphagia, regurgitation, epigastric pain, constipation, and distention). No correlation was found between esophageal motility alterations and gastrointestinal symptoms, nor with the clinical stage of disease in accordance to Brook (r = 0.27). CONCLUSION: These results show that patients with DMD present esophageal motor disorders in both striated and smooth muscle, as well as upper and lower gastrointestinal symptoms. Specialized motility studies could yield a better understanding of disease, and, possibly with adequate treatment, provide for a better quality of life in children with DMD. PMID- 9339238 TI - Effect of embryo density and growth factors on in vitro preimplantation development of mouse embryos. AB - Embryo development depends on maternal and embryonic factors that may regulate genetic programs in early development. Effects of growth factors on proliferation, differentiation and morphogenesis along embryogenesis have been documented. However, studies have not established the role of growth factors in the preimplantational period. The purpose of this study was to investigate the possible effects of growth factors and embryo density on mouse preimplantation development in vitro. Two- and eight-cell CF-1 embryos were cultured individually or in groups of ten in HTF medium, alone or with EGF, TGF-beta 1 and IGF-I. Cleavage rate varied greatly with growth factors and increased significantly when eight-cell embryos were cultured in groups. On the other hand, when two-cell embryos were cultured in groups, the cleavage rate was slower than that obtained when embryos were individually cultured. The differentiation rate increased significantly in two-cell embryos cultured in groups (p < 0.05). EGF, TGF-beta 1 and IGF-I increased differentiation rates significantly in two-cell embryos individually cultured for 68 hours. The combination of EGF and TGF-beta 1 increased the differentiation rates significantly. The other combinations were not effective in modifying this parameter. Hatching rates increased in embryos cultured in groups (p < 0.05). TGF-beta 1 decreased this parameter significantly in two- or eight-cell embryos cultured in groups (p < 0.05). The data described in this report suggest that preimplantational mouse embryos produce some factor or factors that enhance its development, specially the differentiation and hatching rates. However, a functional role for polypeptide growth factors during preimplantational development has to be determined. PMID- 9339239 TI - Effect of diphenylhydantoin administration on the growth of the functional components of rat skull. AB - With the purpose of studying the effect of diphenylhydantoin (DPH) administration on the growth of the functional components of the rat skull, male Wistar rats weighing 61.6 +/- 0.6g were assigned to 1 out of 4 different groups. One of them received saline and was taken as normal control. The others were injected once daily with 25, 50 or 100 mg/kg of b.w. of DPH i.p. for 20 days. Another group of rats was put under a restricted diet (RD) (20% of normal intake) for the same time for evaluation of the cranial dimensions. On day 21 the rats were killed by ether overdose and fifteen cranial dimensions were evaluated as previously described employing Pucciarelli's method. The body weight gain of DPH injected rats was up to 20.7% lower independently of drug dose. The rats of the RD group showed a similar reduction. The amount of food consumed by DPH rats was 16% lower than that consumed by controls independently of drug doses. The concentration of alkaline phosphatase and hemoglobin in rats treated with 50 or 100 mg/kg b.w. of DPH was lower than in controls and RD-animals. However, urea and total calcium in plasma were unchanged in DPH-treated rats as compared to controls. Mean appetite quotient, efficiency of protein and energy utilization did not appear to change in response to the treatment with DPH or maintained under a restricted diet. The cranial dimensions of rats, injected with 25 mg/kg b.w. of DPH were not statistically different from those of the control and RD-groups. When the dose of DPH injected was 50 mg/kg b.w. the neurocranial width and height and the spachnocranial length were significantly lower than controls and RD-values. Moreover, all the dimensions corresponding to neurocranium and splachnocranium (width, height and length) of the rats injected with 100 mg/kg b.w. of DPH were significantly lower than those of control and RD-groups. The disharmonius growth of the skull do not appear to be dependent on suboptimal energy intake, efficiency of protein, energy utilization renal failure and anemia. PMID- 9339240 TI - Action potential duration and contraction after rest at room temperature in guinea pig papillary muscle. AB - Simultaneous recordings of action potential and isometric tension of right papillary muscles were performed. After regular stimulation at 1 Hz, pauses of 10 min were allowed. In the first beat after rest, we measured action potential duration at the 90% of the repolarization (APD90), maximal twitch tension (T), time to peak of contraction (TTP), and rate of development of tension (+dT/dt) and relaxation (-dT/dt). Values were normalized against pre-rest ones. No significative changes were observed after rest at 35 degrees C. After rest at 25 degrees C APD90 and TTP were prolonged but T was reduced. Post-rest +dT/dt were slower, dT/dt did not show significative changes. Nifedipine 10 microM prevented post-rest APD90 lengthening, and produced a further reduction of mechanical response. Substitution of external Na+ by Li+ shortened APD90, increased T of either regular or post-rest beats and led to calcium overload signs. When pause were allowed during Na+ substitution, calcium overload signs were attenuated. We conclude that the combination of rest and room temperature diminished [Ca++]i mainly by Na+/Ca++ mechanism. The reduction of [Ca++]i in turn could delay the inactivation of iCa. As a consequence, longer APs were obtained, accompanied by weaker and slower mechanical responses. Changes in TTP and + dT/dt could suggest that post-rest contractions in room temperature, are dependent of extracellular Ca++ rather than a deplected RS. PMID- 9339241 TI - Effect of different gonadal states on the forskolin stimulated adenylate cyclase activity in rat brain. AB - Forskolin-stimulated adenylate cyclase activity, measured in the hypothalamus and cerebral cortex differs in male and female rats. The gonadal steroid treatment performed induced changes in the studied adenylate cyclase activity probably in relation to the sex of the animals. The stimulated-forskolin adenylate cyclase activity in the hypothalamus from orchidectomized males showed more sensitivity than ovariectomized females. Finally, in male rats, the effects of castration on the hypothalamic enzymatic activity were partially restored by the administration of testosterone dipropionate. On the other hand, estradiol decreased the forskolin-adenylate cyclase activity in the female hypothalamus and cerebral cortex. The results show that the forskolin-stimulated adenylate cyclase activity may be related with the sex and/or the gonadal state of experimental animals. PMID- 9339242 TI - Proliferative and maturative behaviour patterns on murine bone marrow and spleen erythropoiesis along hypoxia. AB - The present study was performed to determine quantitative and qualitative effects of hypoxia on murine erythron. CF1 mice were submitted to hypobaric hypoxia (HH) along 18 days. The proliferative response to recombinant human erythropoietin (rHuEPO: 0-250 mU/ml) was analyzed by DNA assays from bone marrow and spleen cells at different times. Bone marrow proliferative response showed a slight increment under stress but remained over control by the end of the experience. Splenic erythroid proliferative response was observed at a maximum rate on day 6 of HH (26 fold) and returned near to control values after day 10. The assessment of erythropoietic maturative pattern was performed by 59Fe uptake assays. Total nuclear cell counts increased in both tissues (1.5 times in marrow and 5 times in spleen) under hypoxia. In addition, percentages of different lineages (erythroid, myeloid and lymphoid) were scored. Total erythroid marrow cell counts increased in a narrowly degree and persisted above basal counts after day 18. Meanwhile, splenic red cells rose to 30 times over control on day 6 and failed sharply near control values from day 12 of HH. Splenic red cells contribution was approximately 60% of total production between 6-8 days. By the end of the assay bone marrow took back erythroid command (90%). These findings indicate correlation between the time course as well as quantitative and qualitative parameters in the patterns of proliferation and maturation. Moreover, the erythron response to hypoxia, seemed to be related to microenvironmental regulations rather than to hormonal variances. PMID- 9339243 TI - Oxidative stress and membrane fluidity in erythrocytes from patients with hemolytic uremic syndrome. AB - In order to investigate the implications of oxidative disturbances in the hemolysis associated with the Hemolytic Uremic Syndrome (HUS), basal levels of lipid peroxidation products, the response to t-butyl hydroperoxide induced damage and membrane fluidity were assayed by the technique of electron spin resonance in erythrocytes spin labeled with 5-Doxyl stearic acid obtained from eight children with HUS, during the 1st, 2nd, 4th and 12th weeks after diagnosis. During the acute phase of the disease, red blood cells (RBC) showed increased initial lipid peroxidation products, a higher susceptibility to oxidative insult and a lower membrane fluidity. All parameters reached control values the 12th week after diagnosis. The results suggest that in the acute phase of HUS, RBCs are exposed to an oxidative imbalance that could contribute to hemolysis directly through oxidative damage and/or by decreasing membrane fluidity. PMID- 9339244 TI - Intracytoplasmatic and extracellular interleukin-1 production by monocytes of lung and colorectal cancer patients. AB - We studied the production of interleukin-1 (IL-1) by peripheral blood monocytes (Mo) from twelve normal subjects (NS) and eight and nine untreated lung and colorectal cancer patients (CP), respectively. No significant changes of extracellular IL-1 biological activity was observed between CP and NS by thymocyte proliferation assay. This result was independent that the cells were treated or not with lipopolisaccharide from E. coli (LPS, 10 micrograms/ml). Moreover, CP present normal amount of antigenic IL-1 beta in LPS treated Mo culture supernatants by enzyme-linked immunosorbent assay (ELISA). The biological activity of IL-1 released was not significant modified after indomethacin (Indo, 10(-6)M) and LPS + Indo treatments. Furthermore, patients showed a low percentage of LPS activated Mo with intracytoplasmatic IL-1 (alpha + beta) compared to normal values. These results were obtained by immuno-alkaline phosphatase staining using monoclonal antibody anti IL-1 (alpha + beta). In conclusion, CP had a reduced number of Mo with intracytoplasmatic IL-1 (alpha + beta) and the difference observed may depend on degradation or in the rate of synthesis of this cytokine. PMID- 9339245 TI - Effects of methylene blue on the development of intrinsic contractile responses of the guinea pig tracheal smooth muscle. AB - The aim of the present study was to investigate the effects of methylene blue, a guanylyl cyclase inhibitor, on the development of intrinsic contractile responses of the guinea pig tracheal smooth muscle. Paired tracheal chains were mounted for isotonic contractions under 500 mg of tension in Krebs-Henseleit solution. The intrinsic contractile tone modulated carbachol-induced contractions and was inhibited by indomethacin, suggesting the involvement of cyclooxygenase products on this tone. Methylene blue (5 x 10(-5)M) irreversibly inhibited the intrinsic contractile responses. Considering that methylene blue prevents any endogenous production of cGMP, it would be expected to enhance the contractions. However, since methylene blue has effects over nitric oxide synthase and nitric oxide itself, we suggest that guanylyl cyclase activation is not important for the development of the intrinsic tone. PMID- 9339246 TI - Guanylate cyclase activity in retina optic nerve and optic chiasm of rats under different illumination conditions. AB - In the present work we have measured the guanylate cyclase activity in soluble fractions from several tissues relevant to the visual response under different illumination conditions. Guanylate cyclase was sensitive to changes of light/dark periods in incubated extract obtained from soluble fractions of retina, optic nerve and optic chiasm. The changes in soluble guanylate cyclase activity found, about 100 fold between dark and light periods in those tissues, indicate a key role for this enzyme. The results showed that light inhibit strongly the soluble retinal guanylate cyclase activity; while it increases the activity of this enzyme in the optic nerve. A generalized photoinhibited response of soluble guanylate cyclase was observed in all studied tissues in prolonged dark adapted animals. The effect of Na+ 1 and 10 mM, and free Ca++ 28 eta M and 2.8 microM on the guanylate cyclase activity was performed in the studied tissues. The enzymatic activity appeared to be inversely related in the retina and optic nerve with regard to the ion exposure, which may involve different ionic control mechanisms. All indicate an active role for the soluble guanylate cyclase in the phototransduction process not only in retina, also in other tissues relevant in the visual response. PMID- 9339248 TI - Bitemporal hemianopia in photosensitive epilepsy: a case study. AB - This paper reports the occurrence of bilateral hemianopia in a 16 year old male who was having unusual seizures accompanied by severe migrainous headaches and loss of vision while watching a television programme and while playing with the computer. Electrophysiological tests not only confirmed his photo and pattern sensitivity, but also showed that he had bitemporal hemianopia. Hence, his basic EEG showed a great deal of abnormality including generalised spike and wave activity which was more marked in the temporal regions. The patient showed classical occipital spikes on exposure to 25 and 50 Hz of intermittent photic stimulation. Pattern sensitivity test evoked photo paroxysmal response within the range of 2-4.5 cycles per degree (cpd). The visual evoked response to binocular flash stimulation produced N2 at 74 ms, P2 at 112-118 ms and N3 at 168 ms. P2 amplitude was 15-17 uV. Monocular right stimulation produced N2 at 72 ms, P2 at 122-124 ms. Monocular left stimulation produced N2 at 82 ms, P2 at 120-124 ms of 14 uV and N3 at 178 ms. Pattern reversal stimulation produced some abnormality. Poor phase reversals were mainly seen to the left occiput with right eye stimulation and poor phase reversals to the right occiput with left eye stimulation. The pattern responses were of normal latency but showed a marked hemispheric asymmetry. The reduction of the response in the left hemisphere with right eye stimulation and the reduction in the right hemisphere with left eye stimulation would suggest the presence of bitemporal field deficit. PMID- 9339247 TI - Multireceptor interactions of haloperidol on rat cerebral frontal cortex "in vitro". AB - As several side effects of neuroleptics would be related to their interactions with several neurotransmitter receptors (R) haloperidol action on muscarinic cholinergic (mACh) R on frontal cerebral cortex preparations was analyzed. Here we show that haloperidol was able to inhibit in a concentration dependent manner the binding of specific mAChR radiolabeled antagonist on cerebral cortex membranes. This effect would be related to its interaction on mAChR of the M1 subtype as haloperidol blocked the stimulation of phosphoinositides (PIs) turnover induced by low concentrations of carbachol similarly as the M1 antagonist pirenzepine. However at high carbachol concentrations haloperidol triggered a potentiating stimulation of PIs hydrolysis that was only blocked by the alpha 1 adrenergic antagonist prazosin indicating an alpha 1 agonistic action of haloperidol on these Rs. These multireceptor actions of haloperidol found "in vitro" would strengthen its association with "in vivo" neuroleptic-induced side effects. PMID- 9339249 TI - [Normal coronarography]. PMID- 9339250 TI - [Normal coronary angiography. Have the indications changed during the 1980's?]. AB - If the indications of coronary angiography are well chosen, the percentage of normal coronary angiographies should decrease. The authors analysed 7858 primary coronary angiographies performed between 1981 and 1990 in patients without valvular or congenital heart disease. The second 5 years were compared to the first. The percentage of primary coronary angiographies decreased (63% vs 75%; p < 0.01), the percentage of women increased (21.7% vs 18.4%; p < 0.001), and the mean age increased (58.5 +/- 0.3 vs 53.9 +/- 0.3; p < 10(-9)). The lesions were less extensive: 16.3% triple vessel disease versus 24.2% (p < 0.001); 31.3% double vessel disease versus 28.1% (p < 0.02); 49.1% single vessel disease versus 44.2% (p < 0.001). The percentage of normal coronary angiographies remained constant: 20.2% in the second five years versus 19.9% in the first. Over the 10 year period, there was no significant difference one year from another. The percentage of normal investigations remained the same in men (15.7%), decreased in women (34.7 vs 40.1%, p < 0.04), remained constant in patients under 60 years of age (24.5 vs 23.8%), but increased in the more elderly (14.9 vs 10.2%; p < 0.001). The percentage remained unchanged in stable angina (19.6 vs 19.8%) and in unstable angina (12.3 vs 11.2%): it increased in cases of atypical chest pain (72.2 vs 54.3%; p < 0.01). Although, globally, the number of normal coronary angiographies was unchanged at 20%, the indications of this investigation were more selective in the younger patients, especially women, in the second five years, but coronary angiography was more commonly performed in elderly patients because of the possibility of benefiting from coronary angioplasty. PMID- 9339252 TI - [Echocardiographic measurement of left ventricular mass associating data of the M and 2D modes]. AB - Calculation of left ventricular mass by M mode echo is based on the assumption that the geometry of the left ventricle is an ellipsoid, the long axis of which is twice that of its short axis. The hypothesis in not always true and often leads to overestimation of the ventricular mass. The authors propose a method combining M mode data (end diastolic dimension, septal and posterior wall thickness) and 2D measurement of the left ventricular long axis: the left ventricular mass was measured by Devereux's and the authors' methods in 185 hypertensives. The 2D measurement of the long axis (mean: 84.7 mm) was much smaller than twice the short axis (mean: 52.3 mm) and the two measurements were poorly correlated. Measurement of the long axis was reproducible. The two methods of calculation were closely correlated (r = 0.95) but, on average, 23% lower with the authors' method. These results seem to be more closely related to ambulatory blood pressure than those of the classical method. The authors' combined method takes into account the true geometry of the left ventricle better than M mode method alone and avoids overestimation of left ventricular mass and the prevalence of excentric left ventricular hypertrophy in hypertensive patients. PMID- 9339251 TI - [Echocardiographic factors predicting the maintenance of sinus rhythm one year after cardioversion for non-valvular atrial arrhythmias]. AB - Echocardiographic factors predictive of the maintenance of sinus rhythm after successful cardioversion were investigated in 94 patients with non-valvular atrial arrhythmias of recent onset. Seventy-five patients with atrial fibrillation and 19 with atrial flutter admitted for reduction of their arrhythmias underwent transthoracic and transoesophageal echocardiography. After excluding a thrombus in the left atrial appendage or checking that it had disappeared (5 patients), and electrical (n = 74) or pharmacological (n = 20) cardioversion was successfully performed. The maintenance of sinus rhythm (n = 44) or recurrence of arrhythmia (n = 50) were controlled every 3 months for one year. The mean value of the peak positive blood flow in the left atrial appendage was 38 +/- 20 cm/s for the whole group. It was not possible to identify an echocardiographic parameter predictive of maintenance of sinus rhythm at one year either in the whole group or in the subgroups with atrial flutter or atrial fibrillation. In the group in atrial flutter, the mean value of the peak positive blood flow in the left atrial appendage was significantly greater than in the group with atrial fibrillation: 49 +/- 22 cm/s vs 35 +/- 18 cm/s, respectively; p < 0.05. The peak of positive flow in the left atrial appendage was statistically related to indirect parameters of left atrial function and of left ventricular function in the group with atrial fibrillation but only with parameters of left ventricular function in the smaller group with atrial flutter. PMID- 9339254 TI - [Value of tomoscintigraphy with Fourier analysis in the diagnosis of arrhythmogenic right ventricular cardiomyopathy]. AB - ECG gated blood pool tomography has been performed in sixteen patients with right ventricular arrhythmias in whom the diagnosis of arrhythmogenic right ventricular cardiomyopathy was made based on the finding of abnormalities on contrast angiography. They were compared both to control subjects and to patients with primary dilated cardiomyopathy. Thick slices of ventricles were obtained throughout the cardiac cycle in three orthogonal planes: horizontal long axis and short axis thick slices for analysis of right and left ventricular regional wall motion abnormalities and analysis of the spread of the contraction by means of Fourier phase imaging, vertical long axis slices (one for each ventricle) for ejection fractions, because of easy and reproducible determination of valvular planes and analysis of all right ventricular segments, especially the pulmonary infundibulum. Five typical right ventricular abnormalities were seen: decreased ejection fraction (32 +/- 15% vs 55 +/- 3% in control; p < 0.001), increased diameter (ratio of right to left diameters = 1.2 +/- 0.3 vs 0.9 +/- 0.1; p < 0.01), global delayed contraction versus that of the left ventricle (22 +/- 20 degrees vs -2 +/- 6%; p < 0.01), increased dispersion of contraction (32 +/- 16 degrees vs 13 +/- 4 degrees; p < 0.01) and presence of segments with decreased and/or delayed contraction. Right ventricular disease was observed in all the patients: localized form (56%), diffused form (44%). This method provides accurate functional data for diagnosis and follow-up of patients. In future, this wall motion evaluation method may replace planar nuclear angiography as myocardial SPECT have replaced myocardial planar scintigraphy. PMID- 9339255 TI - [Short and medium-term outcome after radiofrequency ablation of the atrioventricular junction]. AB - The aim of this retrospective study was to assess short and long-term mortality and morbidity after radiofrequency ablation of the atrioventricular junction in patients with supraventricular arrhythmias resistant to treatment. This cooperative series included 91 patients (47 men, mean age 68 +/- 9 years). The arrhythmia was paroxysmal in 56% of cases. The mean duration of symptoms was 67 +/- 61 months and 45 patients (49.4%) were in the NYHA Classes III and IV. An escape rhythm was present in 58 patients with a mean frequency of 39 +/- 9/min. Early complications were observed in 5 patients (venous thromboses, pulmonary embolism, mild pericardial effusion and haemothorax). The hospital mortality was 4 patients (4.4%). Death was of cardiac origin in 1 case (cardiac failure). The mean follow-up of patients was 14.5 +/- 8.6 months. Eleven patients died during this period, 8 of cardiac causes: cardiac failure (3 cases), sudden death (3 cases), thrombosis of a valve prosthesis (1 patient) and major electrolyte disturbances causing loss of stimulation in 1 case. Six patients had recurrences. Long-term morbidity was either related to cardiac pacing (3 cases) or cardiac failure (3 cases). Thirteen patients were in NYHA Classes III or IV at the end of follow-up. The authors conclude that radiofrequency ablation at the atrioventricular junction is an effective method of treating resistant supraventricular arrhythmias. Complications are not frequent but mortality appears to be high, probably due to the presence of advanced cardiac disease in the majority of cases. Sudden death seems to be rare and unrelated to pacing defects. PMID- 9339253 TI - [Analysis of frequency-dependence of ventricular repolarisation by the Holter method in young adults. Influence of the autonomic nervous system on the rate dependence of QT]. AB - Adaptation of ventricular repolarization duration to heart rate provides additional information to the static duration of QT interval. METHODS: QT/RR relation and his slope were determined using 24-hour ECG recordings from 17 young male normal volunteers (mean age: 22 +/- 3 years). In order to determine the influence of the autonomic nervous system on the rate-dependence of QT, the authors compared the slopes obtained from recordings during the day and the night. Then, the respective rate-dependences of the entire QT interval (QTe) and its initial subdivision (QTa) and their correlations with parameters of heart rate variability were studied. RESULTS: the QTa/RR and QTe/RR relations were constantly linear, with very significant regression coefficients. The daytime QTa/RR and QTe/RR slopes were significantly steeper than the nighttime ones (0.138 +/- 0.035 vs 0.108 +/- 0.040, p < 0.05 for QTe/RR, 0.160 +/- 0.069 vs 0. 108 +/- 0.055, p < 0.01 for QTa/RR). The early part of QT showed a stronger rate dependence than the global QT, but only at daytime (0.160 +/- 0.069 vs 0.138 +/- 0.035, p < 0.05). No correlation was found between rate dependences and heart rate variability parameters. CONCLUSION: the authors demonstrate, from ambulatory ECG recordings, the influence of the autonomic nervous system on the rate dependence of ventricular repolarization in normal young adults, and a difference in rate-dependence between the entire QT interval and its initial part QTa, due to differences in autonomic nervous system tone. This heterogeneity should be taken to account in the study of pathological changes or drugs effects on ventricular repolarization. PMID- 9339256 TI - [Application of a new covered endoprosthesis in the treatment of occlusive and aneurysmal peripheral arterial diseases]. AB - The aim of this study was to report the authors' experience of a new auto expandable nitinol stent covered by a thin layer of polyester, the Cragg Endopro System 1, for percutaneous internal revascularisation. One hundred and forty-two patients (120 men, 22 women; average age: 63.5 +/- 10 (38-88) received a total of 204 stents (58 iliac, 75 femoral, 9 politeal). The lesions were stenosis in 61 cases, occlusions in 61 cases and aneurysms in 20 cases. The mean length at the femoro-popliteal level was 14.2 +/- 2.4 cm (4-30), at iliac level 9.4 +/- 0.9 (3 15). Implantation was successful in 140/142 cases, a technical success in 136/142 cases (96%). There were 4 acute thromboses requiring surgery and 4 others treated successfully by thrombolysis. There were 18 secondary thromboses. Twenty-nine patients had pyrexia and pain in the treated limb for several days. Over a 27 months follow-up all the iliac stents remained patent; there were 8 restenoses unrelated to the stent, 7 of which were treated by a repeat angioplasty. The primary (PI) and secondary (PII) patency rates at 27 months were: iliac, PI = 97%, PII = 100%; global femoral, PI = 64%, PII = 76% (stenosis PI = 59%, PII = 81%; occlusions, PI = 65%, PII = 74%); lesions of less than 15 cm, PI = 68%, PII = 93%; lesions over 15 cm, PI = 54%, PII: 76%; aneurysms, PI and PII = 88%). The authors conclude that the Cragg Endopro System 1 stent is safe and effective in the treatment of long lesions and aneurysms with encouraging medium term results suggesting that it may be a real alternative to surgery. PMID- 9339258 TI - [Effects of active synchronized prolonged coronary perfusion in the animal]. AB - The efficacy of a system of active diastolic synchronised coronary perfusion was studied during prolonged balloon angioplasty in 8 sheep. In the first part of the study (group 1) including 5 animals, the aim was to study the effects of high and constant flow (48 ml/min) for 90 minutes perfusion on haemolysis, the arterial wall and the perfused myocardium. The second part of the study (group 2), including 3 animals, assessed whether flow adapted to the extent of the vascular bed perfused (24 to 40 ml/min) could protect the myocardium for an interval of 60 minutes. In group 1, after 90 minutes of perfusion (48 ml/min), there was no haemolysis, or jet lesion of the arterial wall distal to the catheter tip. On the other hand, the creatinine phosphokinase levels increased at the 60th minute (188 vs 119 i.u./l for controls) and at the 90th minute (238 vs 119 i.u./l; p < 0.05). Moreover, the perfused myocardium was the site of histological lesions. These observations showed myocardial changes due to the "overflow phenomenon". In group 2, the flow rate was adapted to each animal, increasing progressively until disappearance of electrocardiographic signs of ischaemia (ST elevation) and maintained for 60 minutes. No signs of haemolysis, jet lesions or myocardial changes were observed, with absence of creatinine phosphokinase elevation and histological abnormalities. These preliminary results show that the system investigated allowed myocardial protection after arterial occlusion for an interval of 60 minutes. PMID- 9339257 TI - [Can 1/1 atrial flutter be foreseen by class I anti-arrhythmics?]. AB - 1/1 atrial tachycardia or "quinidine" flutter under class I antiarrhythmic drugs is a serious complication of these agents which, unfortunately, cannot be anticipated. The aim of this study was to review the cases of 11 patients who had suffered this complication of class I antiarrhythmic therapy to see if it could have been prevented. All drugs of this class were included. The 11 subjects were aged 57 to 78: 7 had no apparent underlying cardiac disease and the others had valvular (n = 1), hypertensive (n = 1) and ischaemic (n = 2) heart disease. They were treated for episodes of paroxysmal atrial fibrillation or tachycardia. In the absence of treatment, 7 patients had a short PR interval on the ECG (PR between 0.11 and 0.14 s). In the other 4, the PR interval was normal (0.16 to 0.20 s), but the P wave was widened with appearances of left atrial hypertrophy or an intra-atrial conduction defect. High amplification ECG performed in 3 patients showed continuity of atrial and ventricular depolarisation. Atrial stimulation showed excellent nodal conduction with a Wenckebach point of 200/min. The authors conclude that a short PR interval is predisposing factor to 1/1 atrial tachycardia with class I antiarrhythmics. High amplification ECG which allows identification of the end of the P wave with respect to the QRS complex could help identify subjects at risk when the P wave is widened and that, consequently, the PR interval appears to be normal. PMID- 9339259 TI - [Anti-streptokinase antibodies]. AB - The use of thrombolytics has significantly improved the management of patients with acute myocardial infarction but available molecules remains imperfect: restoration of normal coronary patency in about half the cases, risks of reocclusion, risk of bleeding. Streptokinase (SK), which is the least expensive agent, has disadvantages as do the other thrombolytics. SK, an immunogenic bacterial protein, has another feature: administration of SK leads to an immunitory response with the production of specific anti-streptokinase antibodies. These anti-streptokinase antibodies may interfere with the future administration of a compound containing SK either by inducing an allergic response or by neutralising the SK and making it ineffective. Moreover, anti streptokinase antibodies, the result of previous streptococcal infections, are present in the circulation. Although the prevalence of anti-streptokinase antibodies in the general population, especially coronary patients at risk of myocardial infarction, is not well known and their potentially harmful effects are even less well known. In particular, the platelet aggregant effect in vitro of these antibodies in the presence of SK was not taken into account in the trials studying the influence of anti-streptokinase antibodies in the results of thrombolysis by SK. The anti-lytic effect of anti-streptokinase antibodies is well documented. For this reason, it is not recommended to readminister SK in patients who have previously been thrombolysed with a product containing SK. PMID- 9339260 TI - [Epidemiological course of cardiovascular diseases in developing countries]. AB - Though the environmental and medical conditions are very different, similar population characteristics can be observed in the developing countries. The mean age of the population is young. Most people have a rural way of life, but migrations towards towns result in a disorganized urbanization and in habits more predisposing to cardiovascular diseases. Care access is often difficult for the patients. With respect to risk factors, smoking is increasing, hypertension is highly prevalent and severe, a trend towards obesity is frequent in medium or high economical level people. S or C hemoglobin diseases seem to be associated with coronary heart disease. In spite of very insufficient statistical data, it appears that: cardiovascular disease mortality is increasing when total mortality is decreasing: ischemic and hypertensive heart diseases are increasing when streptococcal or nutritional heart diseases are stabilizing or decreasing. The authors seem to be the different developing countries in respect to the crossing of these curves. Some countries have not reached the crossing. Subsaharan Africa for instance. Others have gone beyond the crossing, some Asian countries for instance. Other countries seem to be at the intersection (Mediterranean or Latin American countries). But many countries suffer the double burden of increasing and decreasing diseases. There is a general lack of prevention owing to other competing priorities and also to economical, social and educational difficulties. However in some developing countries feasibility and efficacy of preventive measures have been proved. PMID- 9339261 TI - [Heart valve diseases specific in rheumatoid polyarthritis. Apropos of 2 cases]. AB - Rheumatoid nodules represent a rare cardiac valvular involvement in rheumatoid arthritis. Patients are usually asymptomatic. We report two cases of such involvement: one presented as a tumour implanted on the mitral valve, with systemic embolisation; the other presented as aortic regurgitation with acute heart failure. Surgical treatment was performed in both cases. Histological examination revealed typical rheumatoid nodules. The authors discuss valvular involvement in rheumatoid arthritis. PMID- 9339262 TI - [Late discovery of inferior vena cava draining into the left atrium after surgical closure of atrial septal defect]. AB - Drainage of the inferior vena cava into the left atrium during surgery for closure of an atrial septal defect is a rare complication. More common in low situated defects, it was more frequent when this type of surgery was performed without cardiopulmonary bypass. This diagnosis was made in a 45 year old woman with cyanosis operated 28 years previously. The right-to-left shunt was demonstrated by the hyperoxia test and confirmed by perfusion pulmonary scintigraphy and contrast echocardiography but only when the contrast was injected in the inferior vena cava territory, and by angiography. The surgeon confirmed the abnormality, closed the interatrial septum and reconnected the inferior vena cava to the right atrium. PMID- 9339263 TI - [Anatomoclinical study of aortic insufficiency in atrophic polychondritis. Apropos of a case]. AB - The authors report the anatomo-clinical features of aortic insufficiency complicating atrophic polychondritis, a rare inflammatory disease affecting mainly cartilaginous tissues. This case illustrates the inflammatory changes of the aortic wall, particularly progressive during this disease, responsible for aortic insufficiency and aneurysmal dilatation of the ascending aorta which required aortic valve replacement and prosthetic replacement of the ascending aorta. Histological analysis showed inflammatory lesions of the aortic wall comparable to the cartilaginous lesions described in this condition and suggesting a common physiopathogenic mechanism. PMID- 9339264 TI - [A rare cause of inappropriate shocks of an implantable automatic defibrillator. Twiddler syndrome]. AB - The authors report the case of a patient with an automatic defibrillator implanted by an endocavitary approach. The device emitted a series of inappropriate shocks. They were triggered by the detection of myopotentials resulting from lesions of the lead due to Twiddler's syndrome. This was reproduced by telemetry in real time and confirmed by chest and abdominal X-ray and the peroperative findings. Treatment consisted of ablation of all implanted material which was replaced by a new retropectoral model. PMID- 9339265 TI - Behavior as an efficacy outcome. PMID- 9339266 TI - Changes in neuropsychiatric symptoms as outcome measures in clinical trials with cholinergic therapies for Alzheimer disease. AB - Alzheimer disease (AD) is a multifaceted disorder with primary manifestations involving neuropsychologic, neuropsychiatric, and neurologic domains. These primary disturbances are based on brain dysfunction and produce secondary effects on patient activities of daily living and quality of life, and have adverse consequences for caregivers, family members, and society. Apathy, agitation, mood disturbances, irritability, disinhibition, delusions, aberrant motor behavior, and abnormalities of sleep and eating are common in AD. The neuropsychiatric disorders of AD are mediated by limbic system and frontal lobe dysfunction and are related in part to the cholinergic deficiency affecting these structures. Cholinomimetic therapy ameliorates the behavioral and emotional disturbances of AD. Cholinergic compounds are unique psychotropic agents that exhibit disease specificity, exerting beneficial effects only in diseases such as AD with cholinergic deficits. Cholinergic agents are potentially useful in the treatment of behavioral disturbances of AD, and their neuropsychiatric effects should be assessed in all clinical trials of these compounds. PMID- 9339267 TI - Behavioral outcomes in clinical trials for Alzheimer disease. AB - The use of behavioral scales is an important component in determining efficacy of new drugs in clinical trials for Alzheimer disease (AD). Behavioral assessment in clinical trials must be sensitive to disease heterogeneity, disease progression, and drug modification of behavior. Three such scales, the Behavior Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD), the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) Behavior Rating Scale for Dementia (C BRSD), and the Cohen-Mansfield Agitation Inventory (CMAI), are useful in clinical trials. The BEHAVE-AD reliably assesses the severity of a range of AD symptoms (7 areas with 25 items) and rates behavioral impact on caregivers. The C-BRSD enables reliable assessment of the frequency of behaviors (8 areas with 48 items) in AD and monitors relevant behaviors throughout the course of the disease. However, it does not assess the impact of behaviors on caregivers. The CMAI focuses on assessment of agitation and aggression and is compatible with C-BRSD but does not assess the impact of agitation on caregivers. A recent trial evaluated the C-BRSD and the CMAI in more than 300 AD and normal elderly individuals. Both of these scales discriminated between AD and non-AD patients, were sensitive across disease severity, and could track behavioral changes over 12 months of AD progression. The BEHAVE-AD, C-BRSD, and CMAI scales are valid, reliable, rapid to administer, cover relevant behaviors occurring during the course of the disease, and are appropriate for use in AD clinical trials. PMID- 9339268 TI - The selective muscarinic agonist xanomeline improves both the cognitive deficits and behavioral symptoms of Alzheimer disease. AB - The therapeutic effects of selective cholinergic replacement using oral xanomeline, an m1/m4 receptor agonist, were assessed in a multicenter study of 343 patients with Alzheimer disease (AD). Patients were randomized to parallel treatment arms (placebo, 25, 50, and 75 mg t.i.d. xanomeline) and followed through 6 months of double-blind therapy and 1 month of single-blind placebo washout. Completer analysis, using the cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-Cog), revealed a significant treatment effect (75 mg t.i.d. vs. placebo; p = 0.045). Similar assessment of global status, using the Clinician's Interview-Based Impression of Change, was also significant (75 mg t.i.d. vs. placebo; p = 0.022). Treatment Emergent Signs and Symptoms analysis of the Alzheimer's Disease Symptomatology Scale, revealed highly significant (p < or = 0.002) dose-dependent reductions in vocal outbursts, suspiciousness, delusions, agitation, and hallucinations. On end-point analysis, the Nurses' Observational Scale for Geriatric Patients also showed a significant dose-response relationship (p = 0.018). The improvement in ADAS-Cog provides the first clinical evidence of involvement of the m1 muscarinic receptor in cognition. Furthermore, the favorable effects of xanomeline on disturbing behaviors suggest a novel approach for treatment of the noncognitive symptoms of AD. Although adverse effects (mainly gastrointestinal) associated with the oral formulation appear to limit its use, a large-scale study investigating the safety and efficacy of transdermal xanomeline is under way. PMID- 9339269 TI - Pharmacologic challenges in Alzheimer disease and normal controls: cognitive modeling in humans. AB - Alzheimer disease (AD) is a progressive disorder characterized by cognitive and behavioral dysfunction, central to which are deficits in the cholinergic and other neurotransmitter systems. These results in the essential symptoms of dementia, including impairment of memory, judgment, and abstract thinking. The pharmacologic relationships among the various neurotransmitters (e.g., cholinergic, serotonergic, nicotinic, and dopaminergic) are highly complex and are still being investigated. Information on the pharmacologic basis of cognitive and behavioral dysfunction in AD has applications to drug therapy. One method of obtaining this information is by pharmacomodeling, using individual or combined drugs. Joint cholinergic antagonism with both muscarinic and nicotinic blockade combines to produce short-term memory impairment, which approximates to mild AD in normal elderly people. This effect is better than that achieved with either agent alone. Mixed cholinergic and serotonergic antagonism has an effect on the cognitive function of AD patients and on depression-related behavior. Dopaminergic dysfunction is linked with the development of hallucinatory and psychotic symptoms and may also be involved in dysfunction of verbal fluency. Combination pharmacomodeling allows the various behavioral and cognitive deficits in AD to be studied and allows models for drug trials to be developed. PMID- 9339270 TI - Behavioral changes associated with different apolipoprotein E genotypes in dementia. AB - Behavioral dysfunction is a problem in patients with Alzheimer disease (AD), and is apparent in up to 67% of individuals. Such changes are a primary cause of individual institutionalization and often lead to their functional disability. As AD progresses, the worsening of behavioral dysfunction becomes increasingly evident and is linked with decreased patient survival. Unfortunately, some of the more common drug therapies used in AD patients to stabilize other facets of their disease worsen behavioral dysfunction. Behavioral changes are associated with endogenous and exogenous factors such as disease stage, environmental factors, other medical conditions, drug regimen, and AD genotype. The most commonly examined and important genotype in AD is the apolipoprotein E (APO E) series, and APO E genotyping is also a useful diagnostic tool. The most frequent APO E genotypes encountered in AD are APO E-4/4, APO E-3/4, and APO E-3/3. In the current study, AD behavioral dysfunction, anxiety, and psychoses were commonly associated with the APO E-3/3 genotype, whereas disorientation, agitation, depression and motor disorders were common among patients with the APO E-4/4 and APO E-3/4 genotypes. These differences were not statistically significant but they suggest that different APO E genotypes influence the phenotypic expression of specific noncognitive symptoms, including behavioral function, in AD. PMID- 9339271 TI - Behavior and caregiver burden: behavioral problems in patients with Alzheimer disease and its association with caregiver distress. AB - Behavioral dysfunction in Alzheimer disease (AD) is a major influence on the morbidity and disability of patients and is central to decisions on patient institutionalization. Behavioral dysfunction ranges from withdrawal, apathy, and depression to hostility, anger, and aggression, with most patients exhibiting some symptoms during the course of the disease. Symptoms of depression are common in AD patients (17-30%) and are associated with broad behavioral dysfunction and increased functional disability. Furthermore, the occurrence of depression in patients correlates strongly with caregiver burden and depression. This report summarizes the relationship between caregiver distress and patient behavioral problems. Administration of the Revised Memory and Behavior Problem Checklist provided information on the frequency of behavioral problems and their association with caregiver distress. In one study of 201 patient-caregiver dyads, depression-related behaviors were confirmed as the most distressing to caregivers. In another, the rates of caregiver depression were high (75%) among those caring for clinically depressed AD patients. Indeed, in a third study, 100% of patients with depression had depressed caregivers. The vulnerability of caregivers to depression is linked to their own age, gender, physical ability, personality, and available social supports. Alleviation of caregiver distress, burden, and depression will be of great value in the improvement of AD patient care. PMID- 9339272 TI - Informant-rated activities-of-daily-living (ADL) assessments: results of a study of 141 items in the U.S.A., Germany, Russia, and Greece from the International ADL Scale Development Project. AB - The analyses of an international pilot study presented in this article focused on the development of a cross-nationally valid activities-of-daily-living (ADL) scale sensitive to therapeutic effects in patients with mild memory impairment and mild to moderately severe dementia. The present report, which is part of an ongoing international research project, describes field testing results for 141 informant-rated items. The comparability of samples investigated in research centers in the U.S.A., Germany, Russia, and Greece concerning cognitive decline was evaluated globally as well as psychometrically using the Global Deterioration Scale, the Short Cognitive Performance Test, and the Mini-Mental State Examination. In the participating countries, a composite ADL score discriminated well between different stages of cognitive impairment because of dementia. However, international differences between the applied measures were observed. A practical ADL scale showing therapeutic sensitivity and international utility, will be constructed from the 141 informant-rated items under investigation in this pilot work. PMID- 9339273 TI - Mammographic visualization of a nonpalpable breast mass through a radiolucent breast implant. AB - We report a woman who underwent augmentation mammaplasty as part of a clinical trial of the Trilucent (soybean oil-filled) breast implant in Genoa, Italy. Five months after surgery a mammography was performed in response to the patient's complaints of pressure and tightness in the area of her left breast. The mammogram clearly demonstrated a 5-mm fibroadenoma of the left breast. This is the first documented case of a nonpalpable breast lesion that was detected by mammography through the new radiolucent, triglyceride-filled implant. PMID- 9339274 TI - Effects of capsular contracture on ultrasonic screening for silicone gel breast implant rupture. AB - Unlike computed tomography and magnetic resonance imaging, ultrasound is an inexpensive test of potential use in detecting silicone gel breast implant (SBI) rupture. However, periprosthetic capsular contracture can make ultrasonic diagnosis of rupture difficult because the contracture-related radial folds inside the SBI can lead to a false-positive diagnosis of rupture. This study was conducted to determine the effects of capsular contracture on the ability of ultrasound to diagnose SBI rupture. Preoperative ultrasonic results of 122 SBIs were compared with surgical findings at the time of implant removal. The sensitivity and negative predictive values of ultrasound were lower in the presence of a contracted capsule (41.2% vs. 68.7%, p = 0.062; and 58.3% vs. 79.6%, p = 0.056 respectively). Ultrasound should be considered reliable in diagnosing SBI rupture only in the absence of a contracted capsule. PMID- 9339275 TI - Postoperative infection following clean facial surgery. AB - Postoperative facial wound infection and breakdown adds to patient morbidity and compromises the cosmetic outcome. This prospective study of 351 patients with a total of 464 wounds for clean elective facial surgery assessed three significant risk factors for wound infection: operative site, oncological surgery, and complex surgery. The findings demonstrated a significantly higher infection rate for the nasal and auricular zones compared with the rest of the face. Wound infection rates were 6.5% for the nasal area, 5% for the auricular area, and 1.5% for the rest of the face. The higher risk of infection to these zones was found to be independent of the lesion excised (benign vs. malignant) and the complexity of the surgery performed. Oncological surgery (skin cancer) and complex surgery (skin grafts and local flaps) were found to increase the risk of postoperative infection significantly by up to fifteenfold compared with nononcological operations with direct closure. The associated morbidity and compromised cosmetic results with facial surgical wound breakdown makes it important to identify these higher risk factors and consider added prophylaxis. PMID- 9339276 TI - Complex regional pain syndrome of the breast in a patient after breast reduction. AB - Complex regional pain syndrome (CRPS) is characterized by devastating pain, swelling, and cutaneous discoloration that result from vasomotor dysfunction caused by an abnormally accelerating sympathetic loop reflex after trauma or surgery. Although in the extremities CRPS is well documented as reflex sympathetic dystrophy, it only has been reported anecdotally in the breast after modified radical mastectomy and never reported after breast reduction. We report CRPS in the right breast of a 27-year-old woman after revision breast reduction surgery. The patient had signs of CRPS and symptoms of pain, swelling, epidermal scaling, and cutaneous temperature changes lasting more than 1 year. Liquid crystal thermographic scanning revealed a persistent, clinically significant hypothermic region in the affected breast. Intravenous phentolamine temporarily relieved the symptoms. Subsequent sympathetic blockade of the stellate ganglion alleviated chronic CRPS symptoms. Surgeons should be alert that CRPS may need to be considered in the differential diagnosis of chronic disproportionate pain after breast surgery. Early identification and treatment will help alleviate persistent CRPS symptoms and avoid soft-tissue changes. PMID- 9339278 TI - Resurfacing a circumferentially degloved hand by using a full-thickness skin graft harvested from an avulsed skin flap. AB - Twelve patients with circumferentially degloved hands were treated with full thickness skin grafts harvested from defatted avulsed flaps. All injuries were industrial accidents caused by various roller machines, not crush injuries. Of these 12 patients, 9 patients were degloved from the wrist level and 3 patients were degloved from the forearm. There were 11 distally based skin flaps and one flap was completely detached. Four patients were avulsed distally to the mid palm, with volar neurovascular bundles damaged at the "fenestrae" of the palm, which resulted in devascularization of the involved fingers. Among them, distal fingers were successfully revascularized by microsurgical techniques in 3 patients. The full-thickness skin grafts were prepared from the attached, avulsed skin flap to avoid junctional hypertrophic scarring. The graft was then secured to its anatomic position with multiple skin staples to improve skin graft take. Initial take of the graft averaged 93% (range, 85%-100%). Compared with conventional methods, this approach provides a higher rate of skin take and better cosmetic and functional results. PMID- 9339277 TI - Titanium screw implants for intermaxillary fixation of partially edentulous jaw. AB - Establishment of the best possible relationship between upper and lower teeth is very important when treating jaw fractures and orthognathic deformities in partially edentulous patients. Many surgeons use arch bars and acrylic splints for intermaxillary fixation (IMF) to obtain the best occlusal relationships after the operation. In patients with sufficient teeth, IMF is not so difficult to realize. However, in partially edentulous patients, the available teeth may not be sufficient to apply arch bars or splints. This paper describes a system for IMF of a partially edentulous jaw. Screws made of medical-grade titanium are implanted into the alveolar ridges where two or more teeth are missing. Arch bars or acrylic splints secured on these implants and available teeth can be used safely for IMF. In vitro axial pull-out tests demonstrated that these implants can withstand the traction forces generated by elastics. Five partially edentulous patients, three with mandibular fractures and two with orthognathic problems, were treated with these implants. All patients healed without any complications and with the best possible occlusal relationships. PMID- 9339279 TI - Gorlin's syndrome: the role of the carbon dioxide laser in patient management. AB - The treatment of multiple basal cell carcinomas in patients with Gorlin's syndrome presents a therapeutic challenge. The carbon dioxide laser presents a unique treatment option due to increased surgical speed, a bloodless operating field, reduced postoperative pain and discomfort, and acceptable scars. Six patients with Gorlin's syndrome have been treated with the carbon dioxide laser. Between 14 and 110 lesions were treated in one session under local anesthesia. Pre- and postlaser biopsies of the lesions confirm complete eradication of the tumors. Mean follow-up is 20 months. No local recurrence has been observed. PMID- 9339280 TI - A new in vivo model for the study of fetal wound healing. AB - We have developed a new in vivo model for the study of fetal wound healing. Fetal ICR mice (total gestation, 21 days) received a full-thickness incisional wound in the hind limb at gestational day 14 (N = 100). The wound was made with a 28.5 gauge needle that was passed transplacentally into the amniotic cavity. The wounds were analyzed histologically on postoperative days 0, 1, 3, and 5 by hematoxylin-eosin and Mallory's trichrome stains. Once the wounding technique was mastered, the overall mortality rate for this model was 20% by postwounding day 5. Each fetus healed their wound without scar by postwounding day 3. In 3 animals, 5 microliters of human transforming growth factor beta 1 (25 micrograms per microliter) was injected into the wound site, resulting in scar and an inflammatory cell infiltrate, indicating that the 14-day-gestation fetal mouse can be manipulated if necessary. This model offers the advantages of an in vivo system that can be studied at an early gestational age. Furthermore, it is inexpensive, easy to manipulate, and can be studied with commercially available murine probes. PMID- 9339281 TI - Upper limb reconstruction with reverse flaps: a review of 52 patients with emphasis on flap selection. AB - Reverse flaps lend themselves to transposition from proximal to distal locations in the extremities. This series comprised 18 radial forearm flaps, 17 digital artery flaps, 13 posterior interosseous flaps, 3 lateral arm flaps, 2 dorsal digital flaps, and 1 dorsal metacarpal flap, all of which were utilized in a reverse pattern. The radial forearm flap was mainly chosen for defects involving part of the palm and the palmar aspect of the first web space. The posterior interosseous flap was more commonly utilized for resurfacing the dorsum, dorsal aspect of the first web space, and especially the hypothenar aspect of the hand. The lateral arm flap was used to reconstruct antecubital fossa and proximal forearm defects. All posterior interosseous and lateral arm donor areas were closed primarily. Sensate digital artery flaps yielded 5 mm on average static two point discrimination in 6 to 18 months of follow-up. Functional and cosmetic results concerning the recipient and donor areas were found to be satisfactory. It was concluded that reverse flaps are versatile tools in the coverage of all kind of defects in the upper limb and should be thought of in the first place. PMID- 9339282 TI - A comparison of methodologies for the preparation of human epidermal-dermal composites. AB - The purpose of this study was to compare methodologies for the preparation of human skin composites based on deepidermized acellular dermal matrix, epidermal keratinocytes, and dermal fibroblasts with the aim of preparing a clinically useful skin substitute. Dermal matrices were prepared from normal human skin and we compared methods of sterilization (glycerol treatment, ethylene oxide treatment, and gamma irradiation), methods of removing the epidermis (sodium chloride, phosphate buffered saline, and dispase), and methods of seeding the composites with fibroblasts and keratinocytes. We report protocols for reproducibly preparing composites that share many of the features of normal skin after 7 days culture at an air-liquid interface. Such composites can be based on allodermis pretreated with either glycerol or ethylene oxide (although the latter gave less consistent results than the glycerol treatment). Fibroblast penetration into the dermis could be achieved by culture of cells on the reticular or papillary surface of the dermis. However, we report for the first time that fibroblast entry from the papillary surface only occurred when keratinocytes were also present. PMID- 9339283 TI - First case of surgical treatment of Farber's disease. AB - Farber's disease (Farber's lipogranulomatosis), which is inherited as an autosomal recessive trait, was first reported by Farber in 1952. We report a case of Farber's disease in a 12-year-old female. Her younger brother was affected with Farber's disease and died of it at 2 years of age. When she first presented, our patient's main clinical features were a shrill voice; subcutaneous nodules; contracture of the joints throughout the body; and granulomas of the oral cavity, the pharynx, and the upper and lower eyelids. Serial radiographs disclosed deformation of the joints throughout the body. Due to the granulomas in her oral cavity, she could take little food orally and therefore was malnourished. We performed a granulectomy under general anesthesia, and her difficulty with feeding and upper airway obstruction improved. There is no specific treatment for Farber's disease, and most patients reported have died by 2 years of age. This is the first reported patient with Farber's disease who has been surgically treated. PMID- 9339284 TI - Metastatic malignant blue nevus: a case report. AB - This report presents a 63-year-old Caucasian woman with a malignant blue nevus, which is an extremely rare form of melanoma originating from or associated with a preexisting blue nevus. The background blue nevus on the left upper arm, which had been present for 5 to 6 years, increased in size and darkened in color for 3 months prior to histological diagnosis of malignant blue nevus. Although the tumor looked much like a nodular melanoma clinically, the diagnosis of malignant blue nevus was established histologically. The patient had a poor outcome due to metastatic spread of the tumor to the visceral organs 1 year following the initial excision of the tumor. To distinguish this rare tumor from other melanocytic lesions, strict histological criteria are needed to make the diagnosis of malignant blue nevus. Differential diagnosis includes cellular blue nevus, atypical cellular blue nevus, primary malignant melanoma, and metastatic melanoma to the dermis. Malignant blue nevus is most commonly seen on the scalp. The tumor has an aggressive behavior and metastasizes in the majority of patients. This paper describes the second reported case of malignant blue nevus involving the upper arm. Clinical and histological features of this uncommon tumor are presented, along with a review of the literature. PMID- 9339285 TI - Computer-aided preoperative planning of tissue expansion. AB - The aim of this paper is to introduce a computer program developed to provide objective, quantitative data useful to ensure proper expander selection when having to use a rectangular tissue expander. The program is developed to calculate the volume of a rectangular tissue expander to obtain the exact amount of yield needed to allow for reconstruction of a determined defect. The only data to be supplied are the length and the width of the defect to be reconstructed. In our opinion, although its use is not aimed at replacing clinical judgment based on experience and careful observation, this program may be considered a simple and useful adjunct for the inexperienced surgeon (or the occasional operator) planning to use a rectangular tissue expander. PMID- 9339286 TI - Treatment of scars: a review. AB - Hypertrophic scars and keloids occur as the result of an exaggerated wound healing response of the skin following injury. In addition to presenting a cosmetic concern, hypertrophic scars and keloids may be painful or pruritic and may restrict range of motion. There is no universally accepted treatment modality resulting in permanent hypertrophic or keloid scar ablation. Atrophic scars secondary to surgery, trauma, and common conditions such as acne vulgaris and varicella may also be disfiguring. This review discusses the etiology and clinical course of hypertrophic, keloid, and atrophic scars. The vast array of treatment modalities which have been implemented in an effort to eradicate scars are reviewed. The advent and development of laser technology represents perhaps the most promising treatment modality for the cosmetic and functional improvement of cutaneous scars. PMID- 9339287 TI - A method of reconstructing a natural-looking umbilicus in abdominoplasty. PMID- 9339288 TI - Pathological tissue expansion. PMID- 9339289 TI - Klippel-Trenaunay-Weber syndrome associated with intra-abdominal lymphangioma requiring multiple surgical interventions. PMID- 9339290 TI - Unexpected, late complication of combined free flap coverage and Ilizarov technique applied to legs. PMID- 9339291 TI - Impact of prematurity, stress and sepsis on the neutrophil respiratory burst activity of neonates. AB - The aim of this study was to evaluate the impact of prematurity, sepsis and stress on the neutrophil respiratory burst activity (NRBA) of neonates. For this purpose 122 healthy neonates (89 term and 33 preterm), 33 preterm stressed neonates, 59 septic neonates (12 term and 47 preterm) and 26 healthy adults were studied. The NRBA was assessed after in vitro stimulation by PMA using a whole blood flow cytometric microassay with dihydrorhodamine 123 (DHR 123). It was found that the percentage of responding neutrophils in term neonates was comparable to that found in adults (medians 83.5 and 89.8%, respectively), whereas it was significantly lower in the healthy preterm neonates (median 70.6%, p < 0.05). The NRBA was further depressed in the stressed (median = 63%) and septic neonates, both term and preterm (medians 60.5 and 54.3%, respectively). No correlation with the levels of G-CSF, TNF-alpha and IL-1 beta, which were found to be higher in the stressed and septic neonates, was observed. These findings indicate that prematurity, sepsis and stress significantly depress the respiratory burst activity of neonatal neutrophils. PMID- 9339292 TI - Cord blood IgE against milk and egg antigens. AB - The aim of the present study was to reevaluate the prenatal production of specific IgE for eggs and milk and, in those cases, to determine whether there is a relation to the amount of maternal egg and milk intake. Total and specific IgEs from 160 cord blood-samples were determined by immunoassays using a paramagnetic particle solid phase and an enzyme-mediated chemiluminescent reaction for signal detection. The levels of cord blood IgE for total, egg, and milk were 0.63 +/- (SD) 1.10 IU/ml, 0.020 +/- 0.055, and 0.036 +/- 0.053 IU/ml, respectively. IgE levels specific to egg and milk over 0.03 IU/ml were observed in 33 and 70 out of 160 cases, respectively. To address whether the maternal intake of eggs and milk affects the level of cord blood IgEs, all mothers except 9 were interviewed, and the amount of eggs and milk taken during pregnancy was recorded. No correlation was seen between egg and milk intakes and cord blood IgE levels. Our data demonstrate a high incidence of the prenatal production of specific IgE for eggs and milk which is independent of maternal egg and milk intakes. PMID- 9339293 TI - Effect of post-hypoxic-ischemic inhibition of nitric oxide synthesis on cerebral blood flow, metabolism and electrocortical brain activity in newborn lambs. AB - Since an excessive production of nitric oxide upon reperfusion/reoxygenation may play an important role in post-hypoxic-ischemic (HI) brain injury, we investigated whether immediate post-HI blockade of nitric oxide synthesis by N omega-nitro-L-arginine (NLA) may reduce this injury. In 18 newborn lambs, subjected to severe HI, changes from pre-HI values were measured for carotid blood flow (Qcar [ml/min]) as a measure of changes in brain blood flow, (relative) cerebral metabolic rate of oxygen (CMRO2), and electrocortical brain activity (ECBA) at 15, 60, 120 and 180 min after HI. Upon completion of HI, at the onset of reperfusion and reoxygenation, 6 lambs received a placebo (control group), 6 low-dose NLA (10 mg/kg i.v., NLA-10 group), and 6 high-dose NLA (40 mg/kg i.v., NLA-40 group). Histological damage to cerebellar Purkinje cells was assessed after termination of the experiment. Only the control group showed a distinct initial post-HI cerebral hyperperfusion. From 60 min after HI onward Qcar was decreased to about 75% of pre-HI Qcar in all 3 groups, although none of these changes in Qcar reached statistical significance. Despite the decreased Qcar in all 3 groups, only the control group showed a significantly decreased CMRO2. ECBA and its bandwidth decreased in all groups, but only recovered in the NLA-10 group 180 min after HI. The brain to body mass ratio (%) and percentage necrotic Purkinje cells were, respectively: 15.3 +/- 0.8 and 56 +/- 10 (control group); 12.5 +/- 1.2 and 36 +/- 9 (NLA-10 group), and 11.3 +/- 1.0 (p < 0.05 vs. the control group) and 35 +/- 14 (NLA-40 group). Since post-HI reperfusion injury of the brain has been characterized by a decreased CMRO2 and electrical brain activity, we conclude that preservation of CMRO2 in both NLA groups, but a recovery of ECBA and its bandwidth only in the NLA-10 group, suggests that NLA, and especially low-dose NLA, may reduce post-HI brain injury. PMID- 9339294 TI - Nitric oxide inhibition after hypoxia-ischemia elevates pulmonary arterial pressure and increases oxygen need. AB - Inhibition of nitric oxide (NO) production may reduce post-hypoxic-ischemic (HI) neonatal brain damage, but may also induce pulmonary hypertension by inhibiting endogenous NO production in the pulmonary vascular bed. The aim of this study was to evaluate the effect of nitric oxide inhibition on pulmonary artery pressure and oxygen need after hypoxic ischemia. Severe HI was produced in 18 newborn lambs. After completion of HI the lambs were divided into three groups of 6 animals receiving either placebo (Cont), low dose N omega-nitro-L-arginine (10 mg/kg i.v., NLA-10) or high dose (40 mg/kg i.v., NLA-40) to block NO production. Pulmonary artery pressure (Pap), aortic pressure, blood gases, inspiratory oxygen concentration and ventilator settings were recorded before and 15, 60, 120 and 180 min after HI. Mean Pap rose initially significantly as compared to baseline in all groups at 15 min post-HI, decreased to normal in Cont but not in treated animals; 180 min post-HI mean Pap was significantly higher in both treated groups as compared to control (NLA-10: 32 mm Hg, NLA-40: 34 mm Hg, Cont: 25 mm Hg, p < 0.05 for NLA-10 and NLA-40 vs. Cont). Moreover, in both NLA-treated groups the oxygenation index was significantly elevated 120 and 180 min post-HI as compared to those of the Cont group. NO synthase inhibition after HI causes a prolonged increase in pulmonary artery pressure leading to a higher oxygen need. PMID- 9339295 TI - Bile flow and composition in preterm, term and infant baboons. AB - We studied the maturational changes in bile composition, bile flow and choleretic effects of sodium taurocholate and secretin in preterm (160 +/- 2 days, n = 4, group I), term (184 +/- 2 days, n = 4, group II), 7-day postnatal age (n = 5, group III) and 60-day-old (n = 5, group IV) baboons. The canalicular bile flow was determined by 14C-erythritol clearance. RESULTS: Gall bladder volume increased from group I to group III (0.08 +/- 0.06 to 1.06 +/- 0.93 ml). Bile flow increased significantly from group I to IV (0.13 +/- 0.05 to 0.34 +/- 0.07 microliter/min/g liver weight, p < 0.05). This was associated with significant increases in total bile acid excretion (16 +/- 3.6 to 31 +/- 2.5 mEq/l, p < 0.05). The composition of bile also showed maturational changes with increasing postnatal age. Sodium taurocholate and secretin increased the bile flow significantly in all groups. CONCLUSION: Data from these studies clearly demonstrate that the bile flow and bile acid excretion is immature in preterm and term baboons when compared to 7- and 60-day-old baboons. The present studies also suggest that baboons can be used as a model to study the postnatal maturation of hepatic excretory function. PMID- 9339296 TI - The effect of glucose during ischemia on brain ATP, lactate, and glutamate in piglets. AB - Glucose worsens hypoxic-ischemic brain injury in 0- to 3-day-old piglets. Piglets were randomly assigned to have blood glucose increased with glucose infusion (n = 12), or decreased with insulin (n = 13), or a sham group (n = 10). In the insulin and glucose groups at time 0, both carotid arteries were clamped, and blood was withdrawn to reduce the blood pressure one third. At time 15 min FiO2 was reduced to 6%. At time 30 min cerebrospinal fluid (CSF) was taken for glutamate, and the brains were frozen. The shams had CSF removed and brains frozen without hypoxia or ischemia. Brain ATP was 1.7 +/- 0.09 mumol/g wet weight in the shams, 0.98 +/- 0.09 in the glucose group (p < 0.01 vs. sham), and 0.52 +/- 0.10 in the insulin group (p < 0.01 vs. glucose). Brain lactate levels were 4.3 +/- 1.0 mumol/g wet weight in the shams, 18.3 +/- 1.9 in the insulin group (p < 0.01 vs. sham), and 29.4 +/- 2.6 in the glucose group (p < 0.01 vs. insulin). CSF glutamate was 9.3 +/- 3.6 microM in the glucose group, 9.6 +/- 3.5 in the insulin group, and 2.2 +/ 0.9 in the shams (p < 0.05, glucose and insulin > sham). The Bmax for MK-801 binding was 2.3 +/- 0.2 pmol/mg protein in the glucose group, 2.6 +/- 0.1 in the insulin group (p < 0.05 vs. sham), and 2.0 +/- 0.2 in the shams, and the Kd was the same in the three groups (p = nonsignificant). Brain Na+,K(+)-ATPase was the same in the three groups (p = nonsignificant). Providing additional glucose preserves ATP during hypoxic-ischemic brain injury in the piglet but does not change CSF glutamate or brain-801 binding and probably worsens outcome by elevating brain lactate levels above the threshold for cellular injury. PMID- 9339297 TI - Effects of hyperglycemia on gasping and autoresuscitation in newborn rats. AB - The aim of this study was to test the effects of glucose on the gasping ability and survival in a rat pup model during acute anoxia. Newborn rat pups of both 1 and 8 days of age were given glucose (30 and 60 mg/animal) or saline intraperitoneally and subsequently subjected to anoxia (100% N2). Glucose supplement induced hyperglycemia. Respiration was recorded by barometric plethysmography. The rat pups responded to acute anoxia with a robust sequence of respiratory pattern: hyperpnea, primary apnea, hypoxic gasping and secondary apnea. During anoxia the 1-day-old rats gasped much longer than the 8-day-old rats (23.4 +/- 1.0 vs. 6.1 +/- 0.5 min, p < 0.001). No difference was found in gasping duration between the saline control and the glucose-supplemented 1-day old rat pups. The 8-day-old supplemented rats gasped much longer (9.3 +/- 0.5 min) than the control rats (6.1 +/- 0.5 min, p < 0.01). The animals autoresuscitated when they received oxygen (100%) during the gasping period. When oxygen was given after the gasping period, the survival rate was 33.3% in control and 0% in supplemented 1-day-old rats, and 100% in control and 50% in glucose supplemented 8-day-old rats (p < 0.02). Further controlled experiments for a fixed period of anoxia to 13.5 min resulted in survival rates of 50.0% for controls and 28.6% for supplemented animals, respectively. The overall survival rate was then 85.2% in control and 52.9% in supplemented 8-day-old rats (p < 0.05). Lactate concentration in blood rapidly increased in the first 6 min of anoxia and thereafter gradually increased to 22.1 mmol/l around the last gasp in the 1-day-old rats. Hyperglycemia did not cause further accumulation of lactate despite a transient elevation over the control rats at 6 min of anoxia. In the 8 day-old supplemented animals the lactate level was only modestly increased, probably due to the prolonged gasping period. In conclusion, we found that gasping performance was well preserved in the 8-day-old glucose-supplemented rats, whereas the autoresuscitation mechanism after the last gasp might be altered due to hyperglycemia. In addition, the accumulation of lactate in the blood did not affect the gasping performance and the mechanisms of autoresuscitation. PMID- 9339298 TI - Epilepsy as a "natural laboratory" for the study of human memory. PMID- 9339299 TI - The role of the temporal lobes in recognizing visuospatial materials: remembering versus knowing. AB - Recognition memory for abstract visuospatial designs was assessed in unilateral temporal lobe epilepsy (TLE) patients and normal controls using a remember/know recognition paradigm. Subjects assigned "remember" judgments to recognized items for which they could recall the study presentation, and "know" judgments to items recognized on the basis of familiarity without conscious recollection of the study episode. In Experiment 1 normal controls and left TLE patients gave more "know" than "remember" recognition judgments for visuospatial materials. Right TLE subjects, however, showed the opposite response pattern. Experiment 1a demonstrated that this dissociation between left and right temporal patients occurred in both presurgery and postsurgery patients. In Experiment 2 recognition was assessed following encoding conditions in which subjects answered questions about either the number of lines in the designs or the appropriateness of verbal labels for presented stimuli. The previous pattern of "know" and "remember" responses was replicated for all groups in the line count condition, but was reversed for normal controls in the label condition. These results are interpreted within a theoretical framework in which "remember" responses are based on the contribution of distinctiveness of individual items to recognition whereas "know" judgements reflect perceptual fluency. PMID- 9339300 TI - Examining the right temporal lobe's role in nonverbal memory. AB - Tests of facial recognition and spatial learning were administered to presurgical patients with unilateral temporal lobe EEG foci. Right temporal lobe patients obtained lower facial recognition scores than left temporal lobe patients. The groups performed equally on the spatial learning test. A factor analysis revealed two independent factors: a general visuospatial factor and a more specific facial identification factor. The findings provide support for the existence of two dissociable visual processing systems. Memory impairments associated with right temporal lobe dysfunction may be characterized as an impairment in a ventral visual processing system responsible for facial memory and pattern recognition. PMID- 9339301 TI - A dissociation between perceptual explicit and implicit memory processes. AB - Patient M.S., who underwent right-occipital lobe resection to treat intractable epilepsy, has intact recall and recognition memory for words, but impaired repetition priming in word identification and visual stem-completion tasks. This mirror dissociation to amnesia suggests that explicit recognition and visuoperceptual repetition priming are mediated by distinct neural systems. In prior studies, however, M.S.' recognition memory was tested only with tasks that drew upon his intact verbal knowledge. The present study examined M.S.' recognition memory for nonverbal perceptual information, namely, the modality and font of word presentation and line patterns. M.S.' recognition memory was intact, providing further evidence that perceptual explicit and implicit memory processes are subserved by functionally and neurally independent memory systems. PMID- 9339302 TI - Very long-term amnesia in association with temporal lobe epilepsy: evidence for multiple-stage consolidation processes. AB - The temporal fractionation of long-term retention remains a relatively uncharted area in human memory research, and in particular there is little in the way of neuropsychological data that address this issue. We describe a patient with temporal lobe epilepsy who complained of amnesia for important events that had occurred in the previous 3-24 months, but who reported that her short-term and medium-term memory were normal. She displayed normal performance on traditional tests of short-term and long-term retention, performing at a very similar level to that of age- and sex-matched healthy control subjects on immediate and half hour delayed recall measures. Forty days later, however, she showed a dense amnesia for recall of such information, whereas control subjects could readily recall much of the original stimuli. She also showed evidence of memory loss for news events that had occurred over the previous few years. MRI scanning and EEG brain mapping indicated left temporal lobe pathology, with a possible epileptogenic focus in the left anterior hippocampus. These data provide empirical evidence for the existence of a distinct very long-term consolidation process in human episodic memory and point to its neural correlates in the temporal lobe. Transfer of information into a permanent long-term memory store may entail multiple-stage consolidation processes rather than a single-stage, unitary consolidation process. PMID- 9339303 TI - Accelerated forgetting in association with temporal lobe epilepsy and paraneoplastic encephalitis. AB - The association between epilepsy and amnesia is studied in patient J.T. who presented with a very unusual pattern of memory loss with retention of information for hours to days but rapid forgetting of information that exceeded this time frame. J.T.'s unusual memory profile was studied with several tests administered over week-long intervals of time. There was evidence that his retention decreased in conjunction with increased seizures. During a trial of paraldehyde, a decrease in seizure frequency was associated with enhanced memory. J.T.'s memory problem was unlike that described in prototypical cases of amnesia. His day-long retention of new information alongside his absolute loss of that information days later is consistent with the idea that consolidation is a process that occurs over lengthy periods of time. PMID- 9339304 TI - Memory for emotional words following unilateral temporal lobectomy. AB - We recently reported that patients who had received unilateral temporal lobectomy, including the amygdala and hippocampus, show impaired acquisition in a fear conditioning task (LaBar, LeDoux, Spencer, & Phelps, 1995), indicating a deficit in emotional memory. In the present paper, we examined performance of these patients on two verbal, emotional memory tasks in an effort to determine the extent of this deficit. In Experiment 1, subjects were asked to recall emotional and non-emotional words. In Experiment 2, subjects were asked to recall neutral words which were embedded in emotional and non-emotional sentence contexts. Both temporal lobectomy subjects and normal controls showed enhanced recall for emotional words (Experiment 1) and enhanced recall for neutral words embedded in emotional sentence contexts (Experiment 2). These results suggest that the deficit seen in emotional memory following unilateral temporal lobectomy is not a global deficit and may be limited to specific circumstances where emotion influences memory performance. Several hypotheses concerning the discrepancy between the present studies and the fear conditioning results (LaBar et al., 1995) are discussed. PMID- 9339305 TI - Differential involvement of left temporolateral and temporomesial structures in verbal declarative learning and memory: evidence from temporal lobe epilepsy. AB - A wealth of animal and human research has pointed to a significant involvement of the temporal lobes in memory processing, and yet the different functional roles of temporal cortical vs. mesial structures remain unclear. We studied verbal declarative memory, by using a word list paradigm that differentiates among learning (immediate recall), memory (delayed recall), and recognition, in epilepsy patients being considered for surgical resection of the left temporal lobe. Verbal memory was evaluated preoperatively and during the recording of intracranial event related potentials and postoperatively after selective hippocampectomy, temporal cortical lesionectomy, or anterior two-thirds en bloc temporal lobe resection procedures. Preoperative differences in verbal memory performance as a function of differences in underlying neuropathology, concurrent event-related potentials, and specific patterns of postoperative memory impairments lead to converging evidence that verbal declarative memory relies on a synergistic interaction of at least two functionally distinct brain systems. Material-specific data acquisition, or working memory, is mediated by neocortical temporal structures, whereas long-term consolidation/retrieval is particularly mediated by temporomesial structures. In contrast to the left temporal neocortex, the function of the temporomesial system appears to be material nonspecific. Apparently, its preferential involvement in verbal memory is due to its close interaction with overlying neocortical structures that are specialized for language processing. PMID- 9339306 TI - Reorganization of verbal memory function in early onset left temporal lobe epilepsy. AB - The purpose of this investigation was to examine the issue of reorganization of verbal memory function following early insult to the left mesial temporal region. It was hypothesized that reorganization of memory function was most likely to occur in those patients with an early age of seizure onset who have a more limited degree of extra-hippocampal neuropathology. Fifty-four patients with epilepsy of unequivocal left temporal lobe origin were classified into four groups on the basis of the presence/absence of hippocampal sclerosis and degree of postoperative seizure relief. Measures of verbal learning and memory as well as nonmemory measures were administered both before and 6 to 8 months after anterior temporal lobectomy. Findings were consistent with the reorganization proposal. The clinical and theoretical significance of the findings are discussed. PMID- 9339307 TI - Health impacts of large releases of radionuclides. Physical transport and chemical and biological processes in agricultural systems. AB - The purpose of radioecological models is to make realistic estimates of doses to the public after accidental releases of radionuclides into the environment. Important physical, chemical and biological processes involved in the dispersion and transport of radioactive substances in the atmosphere and along the food chains are presented. The results of the EURAD (EURopean Acid Deposition) model, predicting the deposition patterns of 131I and 137Cs in Belarus and Ukraine after the Chernobyl accident, are discussed. An overview of the most important ecological processes--such as deposition, interception and translocation, weathering, transfers from soil to plants and from plants to animal/animal products, and seasonality in agricultural environments--is given. Examples corresponding to these individual processes, mainly experimental results after the Chernobyl accident and related to radiocaesium and radioiodine, are shown and discussed. PMID- 9339308 TI - Health impacts of large releases of radionuclides. The radioecological significance of semi-natural ecosystems. AB - The transfer of radiocaesium to many food products either produced in or harvested from semi-natural ecosystems is high compared with intensive agricultural areas. Radiocaesium contamination levels in semi-natural foods are highly variable and difficult to predict. Spatial analysis may help to explain some of the variability and give improved estimates of the total output of radiocaesium in food products produced or harvested from semi-natural ecosystems. Consumption of foodstuffs from semi-natural ecosystems can contribute significantly to radiocaesium ingestion by humans. The long effective half-lives that occur for some semi-natural products lead to an increase with time in their importance compared with agricultural products. In determining the importance of semi-natural food products, the diet needs to be considered for both the average population and for special groups who utilize these environments to a greater extent than normal. Effective countermeasures have been developed to reduce radiocaesium levels in some semi-natural products. PMID- 9339309 TI - Health impacts of large releases of radionuclides. Transport and processes in freshwater ecosystems. AB - The partition coefficient (Kd) and the water retention rate (RR) are fundamental components of dynamic, mass-balance models, not just for radionuclides in fresh water but also for contaminants in all aquatic ecosystems. Kd may be regarded as an 'entry gate' and RR an 'exit gate'. Uncertainties in Kd and RR cause uncertainties in model predictions. Uncertainties in important rates for processes within ecosystems (such as sedimentation, diffusion, advection, bio uptake and excretion) cannot be adequately evaluated when uncertainties exist for Kd and RR. Empirical data show that there may be a variation in Kd of two orders of magnitude with environmental factors such as pH. This is important because Kd regulates the amount of radionuclides in dissolved and particulate phases, and hence also pelagic and benthic transport. Pelagic transport is directly linked to the outflow and retention of substances in the water mass, and thus also to concentrations and ecological effects. There are many approaches for sub-models of Kd and RR. Which provide the best predictive power? This chapter gives a brief overview and discussion of the benefits and drawbacks of different alternatives for Kd and RR within the framework of a lake model for radiocaesium. PMID- 9339310 TI - Health impacts of large releases of radionuclides. Radioactive contamination of the marine environment. PMID- 9339311 TI - Health impacts of large releases of radionuclides. Impacts on plant and animal populations. AB - Historical experiments and observations in radioactively contaminated environments have documented radiation impacts on natural plant communities and animal populations. General findings from these studies are reviewed. Despite much information on the response of individual organisms to radiation in the laboratory, the database is more limited and the interpretations more complex for natural populations and communities exposed to radionuclides. These complications are discussed as they pertain to plants and animals in natural environments. Paradigms concerning the recovery of radiation-damaged communities and ecosystems, and areas of needed research are discussed. PMID- 9339312 TI - Health impacts of large releases of radionuclides. Roles of micro-organisms in the environmental fate of radionuclides. AB - Micro-organisms play important roles in the environmental fate of radionuclides in both aquatic and terrestrial ecosystems, with a multiplicity of physico chemical and biological mechanisms effecting changes in mobility and speciation. Physico-chemical mechanisms of removal, which may be encompassed by the general term 'biosorption', include adsorption, ion exchange and entrapment. These are features of living and dead organisms as well as their derived products. In living cells biosorptive processes can be directly and indirectly influenced by metabolism, and may be reversible and affected by changing environmental conditions. Metabolism-dependent mechanisms of radionuclide immobilization include metal precipitation as sulfides, sequestration by metal-binding proteins and peptides, and transport and intracellular compartmentation. Chemical transformations of radionuclide species, particularly by reduction, can result in immobilization. Microbial processes involved in solubilization include autotrophic and heterotrophic leaching, complexation by siderophores and other metabolites, and chemical transformations. Such mechanisms are important components of natural biogeochemical cycles for radionuclides and should be considered in any analyses of environmental radionuclide contamination. Several micro-organism-based biotechnologies, e.g. those based on biosorption or precipitation, are of potential use for the treatment of radionuclide contamination. PMID- 9339313 TI - Health impacts of large releases of radionuclides. The fate and impact of radiocontaminants in urban areas. AB - The Chernobyl accident made it clear that the contaminants released after a severe nuclear accident may spread over large areas, and thereby form a significant external radiation hazard in areas of high population density. Since then, the weathering effects on the deposited radiocontaminants (essentially radiocaesium) have been followed on different types of surface in urban, suburban and industrial areas in order to enable an estimation of the long-term impact of such events. Analytical expressions have been derived for the typical behaviour of radiocaesium on the different surfaces, and dose measurements and calculations for different urban environments have pinpointed which surfaces generally contribute most to the dose and consequently are most important to clean. At this point, after nearly a decade, the dose rate from horizontal pavements has decreased by at least a factor of 10, whereas the dose rate from an area of soil or a roof has generally only been halved. The contamination on walls is the most persistent: it has only decreased by 10-20%. PMID- 9339314 TI - Health impacts of large releases of radionuclides. Internal exposure of populations to long-lived radionuclides released into the environment. AB - This chapter discusses the events that led to the contamination of environments with the long-lived radionuclides of caesium, strontium and other elements, and to the internal exposure of populations living in contaminated areas. Among these events are radioactive releases into the river Techa from the Soviet nuclear weapons facility Mayak in 1949-1956, thermonuclear weapons tests in the 1950s and 1960s, the Kyshtim and Windscale accidents in 1957, and the Chernobyl and Tomsk-7 accidents in 1986 and 1993, respectively. Methods of environmental monitoring and individual internal dose monitoring of inhabitants are described. These are based on measuring the content of radionuclides not only in the air, drinking water and local food products, but also in humans using whole-body counters and analysing excreta and autopsy samples. The dynamics of internal exposure of people of different ages to radionuclides of caesium, strontium and plutonium from the environment are considered. Examples of radionuclide distributions in the environment, and of individual/collective internal doses and related medical effects are presented. PMID- 9339315 TI - Health impacts of large releases of radionuclides. Interactions with human nutrition and other indices of population health. AB - The consumption of food is an important pathway involved in the internal contamination of humans. The site-related critical foodstuffs can be grouped into three main categories: dairy products; aquatic animals, such as fish, molluscs and crustaceans; and other typical foods. The concentration factor plays a more important role than the amount of a certain food consumed. Semi-natural and natural ecosystems are of special interest in this context because they can provide critical pathways for radionuclide transfer to humans, and they can also act as temporary sinks or long-term sources for radionuclides deposited from the atmosphere. From the viewpoint of population health, another important role is played by the countermeasures. The reference values commonly adopted in radiation protection are conservative and they have been established for planning practices that could provide future sources of irradiation. After a large release of radionuclides, the evaluation of the problem must be as realistic as possible, otherwise the countermeasures will imply consequences worse than those produced by the accident itself (without any further intervention). This criterion was clearly stated by the International Commission on Radiological Protection but it was frequently neglected after the Chernobyl accident. The results of a survey on the number of induced abortions following this incident are reported. These suggest that moral and ethical problems are involved above and beyond any economical implications. PMID- 9339316 TI - Health impacts of large releases of radionuclides. Biological effects of prenatal irradiation. AB - After large releases of radionuclides, exposure of the embryo or fetus can take place by external irradiation or uptake of radionuclides. The embryo and fetus are radiosensitive throughout prenatal development. The quality and extent of radiation effects depend on the developmental stage. During the preimplantation period (one to 10 days postconception, p.c.), a radiation exposure of at least 0.2 Gy can cause the death of the embryo. Malformations are only observed in rare cases when genetic predispositions exist. Macroscopic, anatomical malformations are induced only after irradiation during the major organogenesis (two to eight weeks p.c.). A radiation dose of about 0.2 Gy is a doubling dose for the malformation risk, as extrapolated from experiments with rodents. The human embryo may be more radioresistant. During early fetogenesis (8-15 weeks p.c.) a high radiosensitivity exists for the development of the brain. Radiation doses of 1.0 Gy cause severe mental retardation in about 40% of the exposed fetuses. It must be taken into account that a radiation exposure during the fetal period can also induce cancer. It is generally assumed that the risk exists at about the same level as for children. PMID- 9339317 TI - Health impacts of large releases of radionuclides. Late somatic health effects. AB - This chapter reviews the risks of radiation-induced cancer for the dose range likely to occur after releases of radionuclides into the environment. Epidemiological evidence from exposed workers and the atomic bomb survivors of Hiroshima and Nagasaki is surveyed. Influences on such risk functions of individual related quantities (e.g. age, sex, nationality, time since exposure and organs exposed) and of radiation modality-related quantities (e.g. dose, dose rate and radiation quality) are also briefly discussed. PMID- 9339318 TI - Health impacts of large releases of radionuclides. Retrospective radiation dose assessment: an overview of physical and biological measures of dose. AB - Models to estimate population doses from environmental measurements, dietary radioactivity and lifestyle characteristics are useful for populations but difficult to apply accurately to an individual within the population. Individual biological and radiation dosimetry is limited to small numbers of persons and thus has limitations when considering larger groups or populations. Current direct biological indicators of dose are generally limited to doses above 0.1 Gy. New advances in improving the accuracy and sensitivity of these methods offer the promise of validating population estimates and specifying variance in individual doses. An integration of the two approaches will provide the support for more accurate radiation epidemiology and risk assessment. PMID- 9339319 TI - Health impacts of large releases of radionuclides. Cytogenetic effects as quantitative indicators of radiation exposure. AB - Scoring of dicentrics in metaphase preparations of human T lymphocytes is the method of choice for estimating individual whole-body doses of radiation exposure. A quantification of partial-body exposures or non-uniform distribution of the dose is more complicated but it can be achieved by using specific mathematical approaches. For retrospective biodosimetry, conventional scoring of dicentrics is less precise because these unstable aberrations are eliminated with time post-exposure. Symmetrical translocations are not selected against during mitotic division in the haematopoietic cell reproductive centres, so the frequencies of these stable aberrations are generally assumed to remain constant even for decades. They can now be analysed precisely by fluorescence in situ hybridization using whole chromosome-specific DNA probes (chromosome painting) with an alpha-satellite DNA probe for centromere detection. Based on in vitro calibration curves established with single or multicolour paints covering 4-22% of the total human genomic DNA content, scoring of translocations has been applied for dose reconstruction in smaller groups of atomic bomb survivors and victims of the Chernobyl and Goiania radiation accidents. However, prior to routine use, the method requires further validation. Such work includes the precise evaluation of the unexpectedly high frequency of complex exchanges (> or = 3 breaks in > or = 2 chromosomes) found both at > 2 Gy doses of low linear energy transfer (LET) radiation and generally for high LET alpha-particles. Data on the long-term stability of translocations and the appearance of clonal abberrations, as well as improved measurements of the linear coefficient of standard calibration curves, are also required. PMID- 9339320 TI - Health impacts of large releases of radionuclides. Mental health, stress and risk perception: insights from psychological research. AB - Risk perceptions are only slightly correlated with the expected values of a probability distribution for negative health impacts. Psychometric studies have documented that context variables such as dread or personal control are important predictors for the perceived seriousness of risk. Studies about cultural patterns of risk perceptions emphasize different response sets to risk information, depending on cultural priorities such as social justice versus personal freedom. This chapter reports the major psychological research results pertaining to the factors that govern individual risk perception and discusses the psychometric effects due to people's risk perception and the experience of severe stress. The relative importance of the psychometric context variables, the signals pertaining to each health risks and symbolic beliefs are explained. PMID- 9339321 TI - An ECG classifier designed using modified decision based neural networks. AB - In this paper, a neural network based generalized software system is presented for automatic analysis of electrocardiograms (ECGs). The proposed system is capable of intuitively diagnosing the disease from the ECG using the knowledge acquired from the training. A modified decision based neural network which converges in a finite amount of time is employed. The training procedure used automatically varies the size of the network. The system is capable of being trained even without an expert's supervision. The physician can correct the network as and when a misclassification occurs, thus making the system less error prone as time passes. The proposed system has been tested using an ECG data base representing different cardiological conditions such as bundle branch blocks and infarctions. The system is capable of detecting different types of arrhythmias also. PMID- 9339322 TI - Design and implementation of a relational database used in the management of patients with retinoblastoma. AB - Clinical data are most useful in the management of patients with complex medical problems if they are accurate, reliable, and easily accessible by physicians and the medical community at large. Furthermore, the data are most valuable when they can be shared among cooperating institutions. We describe a computer system which exhibits a uniform taxonomy, an integrated on-line dictionary of clinical terms, a coherent temporal layout, and persistent spatial integrity with regard to the values of the variables. The system is user friendly and provides real time data access which can be retrieved by structured query language or may be programmed to be used as part of an international network in the management of patients with retinoblastoma, a malignant and potentially fatal tumor of childhood. Furthermore, because of its design flexibility, this system provides for potential application to other ophthalmic disorders, such as malignant uveal melanoma, and other areas of medicine as well. PMID- 9339323 TI - Vital statistics linked birth/infant death and hospital discharge record linkage for epidemiological studies. AB - A methodology for linking vital statistics linked birth/death data and hospital discharge data is described. The resulting data set combines information on a neonate's sociodemographic characteristics, prenatal care, and mortality aspects and connects it to detailed health outcome and resource utilization data, thus establishing an extensive database for epidemiological studies. In the absence of a universal identifier common to both databases, our linkage strategy relied on using a virtual identifier based on variables common to both data sets. In the case of multiple incidences of the same virtual identifier we used secondary health status information to optimize the likelihood of linking low birth weight or premature infants in one database to infants of similar health status in the other while randomizing cases in which no secondary information was present. Applying our method to the 1992 California birth cohort, we could link 563,114 out of 571,189 eligible births (98.59%). Of these links, 91.2% were established on the basis of unique virtual identifiers. The link was internally consistent and no bias was evident when comparing variable distributions for all single live births in the vital statistics linked birth/death file and linked births in the linked vital statistics linked birth/death and hospital discharge file. Multiple imputation techniques showed that the prediction error incurred by randomization was negligible. Even though computationally intensive, our method for linking the vital statistics linked birth/death file and the hospital discharge file appeared to be effective. However, it is important to be aware of the limitations of the resulting data set, in particular the fact that it cannot be used for tracking individual cases. The method provides a database suitable for a variety of perinatal epidemiological analyses, such as descriptive studies of disease distribution in neonates, studies of the geographic distribution of disease, and studies of the relationship between risk and outcome. PMID- 9339325 TI - Concept locator: a client-server application for retrieval of UMLS metathesaurus concepts through complex boolean query. AB - Concept Locator (CL) is a client-server application that accesses a Sybase relational database server containing a subset of the UMLS Metathesaurus for the purpose of retrieval of concepts corresponding to one or more query expressions supplied to it. CL's query grammar permits complex Boolean expressions, wildcard patterns, and parenthesized (nested) subexpressions. CL translates the query expressions supplied to it into one or more SQL statements that actually perform the retrieval. The generated SQL is optimized by the client to take advantage of the strengths of the server's query optimizer, and sidesteps its weaknesses, so that execution is reasonably efficient. PMID- 9339324 TI - A computer simulation model for the spread of nosocomial infections caused by multidrug-resistant pathogens. AB - A Monte Carlo simulation model was developed for the spread of antibiotic resistant bacteria in hospital units. The model allows for the representation of every patient and staff member. Staff-patient interactions, staff handwashing compliance, admission of colonized patients, and antibiotic use are included in the model. The simulation model provides colonization curves for patients and staff and offers the possibility of simulating different kinds of hospital units. Simulation of the spread of an antibiotic-resistant pathogen in an intensive care unit was performed. We studied the impact of handwashing compliance on colonization. The importance of handwashing in preventing colonization and the influence of admission of colonized patients in perpetuating an epidemic were confirmed by the model. The model offers a new approach to modeling the spread of nosocomial pathogens in hospital units. It allows one to study the impact of infection control measures and represents a valuable educational tool for staff. PMID- 9339326 TI - One or several semantic system(s)? Maybe none: evidence from a case study of modality and category-specific "semantic" impairment. AB - Following cerebral anoxia, EC a 55-year-old patient, exhibited a severe and clear cut pattern of semantic impairments without general intellectual deficit or perceptual difficulty. EC demonstrated a complex neuropsychological picture including a massive visual agnosia and a complete lack of imagery, both of which involved all categories of objects (living and non living) and a category specific word comprehension deficit limited to animal names. Findings are discussed in the light of the theoretical frameworks currently available in the area of neuropsychology. It is argued that neither the single nor the multiple view of semantics fully succeed in providing a satisfactory account of the data and a tentative interpretation of the whole pattern of impairment is proposed in the general framework of non abstractive conceptions of meaning. PMID- 9339327 TI - Thalamic thought disorder: on being "a bit addled". AB - Humans can generate and maintain relatively coherent trains of thought in natural discourse. The neural mediation of this ability and the phenomenology of its breakdown are not well understood. We report a case of a woman with paramedian thalamic strokes involving the mammillothalamic tract, intralaminar nuclei, parts of the dorsomedial and ventral lateral nuclei bilaterally. She presented with a dense amnesia and confusion typical of the syndrome of bilateral paramedian thalamic infarcts. Her Tc-99m HMPAO brain SPECT scan showed decreased thalamic and basal ganglia blood flow. General diminution of cerebral blood flow and areas of further diminution in the right frontal, left temporal and left temporoparietal regions were also observed. Although her amnesia was characteristic of diencephalic amnesia, her most striking clinical feature was a bizarre, disconnected and at times incoherent speech output. Analysis of her speech revealed relatively preserved lexical and morpho-syntactic linguistic production. By contrast, analysis of the macrostructure of her discourse revealed frequent unpredictable topic shifts that were completely unconstrained by contextual factors. Many of her shifts were intrusions from previous topics. We interpret her severely disordered speech output as representing the surface manifestations of a thought disorder (rather than as a language disorder per se) characterized by an inability to maintain and appropriately shift themes that normally guide discourse. Median and intralaminar thalamic nuclei appear to be critical for the neurophysiologic regulation of thalamocortical and striatocortical circuits, which in turn may be critical for the functional regulation of contextually appropriate transitions of thought. PMID- 9339328 TI - Semantic category dissociations: a longitudinal study of two cases. AB - We report the neuropsychological findings of two patients (LF and EA) with herpes simplex encephalitis. Both patients presented a greater deficit for living than non-living categories in a number of tasks, although EA was much more impaired than LF. We controlled the several stimulus variables that might affect the performance and could demonstrate that the dissociation was not artifactual. Neither LF nor EA revealed a selective or preferential involvement of perceptual semantic knowledge, and both showed a homogeneous impairment of perceptual and associative encyclopaedic notions. At a second examination, carried out from 1 to 2 years later, LF showed a good recovery, whereas EA's improvement was confined to the non-living categories. The lesion of both patients affected the left temporal pole and the basal neocortical regions of the left temporal lobe. The involvement of limbic areas was more marked in LF, while the Wernicke area and the posterior parts of the middle and inferior temporal gyri were only involved in EA. Besides the basal temporal areas, also the posterior temporal regions are likely to play a role in determining the clinical picture of such patients, and their prospect of recovery. PMID- 9339329 TI - Deductive and inductive reasoning in Parkinson's disease patients and normal controls: review and experimental evidence. AB - In the present study, fifty-four subjects were tested; twenty-seven with idiopathic Parkinson's disease and twenty-seven normal controls matched in age, education, verbal ability, level of depression, sex and socio-economic status. The subjects were tested on eight tasks. Five of the tasks were the classic deductive reasoning syllogisms, modus ponens, modus tollendo tollens, affirming the consequent, denying the antecedent and three-term series problems phrased in a factual context (brief scripts). Three of the tasks were inductive reasoning, including logical inferences, metaphors and similes. All tasks were presented to subjects in a multiple choice format. The results, overall, have shown nonsignificant differences between the two groups in deductive and inductive reasoning, an ability traditionally associated with frontal lobes involvement. Of the comparisons performed between subgroups of the patients and normal controls concerning disease duration, disease onset and predominant involvement of the left and/or right hemisphere, significant differences were found between patients with earlier disease onset and normal controls and between bilaterally affected patients and normal controls, demonstrating an additive effect of lateralization to reasoning ability. PMID- 9339330 TI - A deficit for arithmetical procedures: lack of knowledge or lack of monitoring? AB - A patient is described with a specific deficit for arithmetical procedures. Unlike in previously described cases, where the observed problems could be attributed to the systematic application of disturbed algorithms, this patient's difficulty seems to stem from an inability to monitor the sequence of operations that calculation procedures specify. Criteria are provided for distinguishing impairments in written calculation due to the application of defective knowledge of the procedures from those determined by lack of monitoring. The role of monitoring and control processes in different calculation components is also discussed. PMID- 9339331 TI - Optic aphasia: evidence of the contribution of different neural systems to object and action naming. AB - Visual stimulus naming was studied in a 66-year-old male patient with optic aphasia subsequent to left occipito-temporal infarction. While having difficulty in naming objects perceived visually, he was able to name objects by viewing gestures illustrating their use, and to name actions shown in pictures. These results suggest that naming performance depends on the kind of stimulus that is visually presented (object vs. action). The present findings lend support to congnitive models which postulate the existence of visual and functional semantic systems. PMID- 9339332 TI - Do individuals with Williams syndrome have bizarre semantics? Evidence for lexical organization using an on-line task. AB - Williams syndrome, a neurodevelopmental disorder, has attracted a great deal of debate concerning the purported intactness of language in the face of other serious cognitive deficits. As more in-depth studies of specific aspects of WS language have emerged, the notion of a preserved language module has been seriously challenged. Although WS vocabulary scores are often impressive, several investigators have claimed the WS semantics are aberrant. All studies hitherto have been based on off-line experiments which necessarily involve metalinguistic processes. This clearly affects the performance of individuals with cognitive deficits. We report here an on-line study probing the semantic structure of the WS lexicon, using a task-semantic priming-which minimises metalinguistic demands. We show that WS subjects display the same taxonomic/category and thematic/functional priming effects as normal controls. The results are discussed in terms of the differences between receptive and expressive language, as well as the fact that although semantic memory and the automatic access to semantic information for individual words is normal in WS, the integration of semantic information into sentence comprehension may be abnormal. The importance of online tasks to highlight such differences is stressed. PMID- 9339333 TI - Knowing where and knowing what: a double dissociation. AB - We report a double dissociation between visuo-spatial abilities and semantic knowledge (knowledge of the names and attributes of objects and people), in two brain-injured people with longstanding stable impairments, using a wide range of tests to explore the extent of the dissociation, MU, who has bilateral lesions of occipito-parietal cortex, shows severe spatial disorientation with relatively well-preserved semantic knowledge. He is contrasted with JBR, who has bilateral temporal lobe damage and shows severe semantic problems and no impairment on visuo-spatial tasks. Our findings thus demonstrate a double dissociation between the performance of semantic and spatial tasks by MU and JBR. This pattern is consistent with Ungerleider and Mishkin's (1982) neurophysiological hypothesis of separable cortical visual pathways; one which is specialised for spatial perception and follows a dorsal route from occipital to parietal lobes, and the other following a more ventral route from occipital to temporal lobes, whose target is semantic information needed in specifying what an object is. PMID- 9339335 TI - Two-dimensional cancellation neglect a review and suggested method of analysis. AB - Directional bias on cancellation has thus far not been standardized. While cancellation tasks are primarily used to assess lateral performance asymmetries, they may also reveal two-dimensional (i.e., combined lateral and radial) neglect patterns. We propose a method to evaluate and report cancellation neglect regardless of whether the neglect pattern is strictly unilateral or two dimensional. Our method establishes the location of the geographic center of all neglected stimuli relative to the page center by averaging their Cartesian coordinates. This "neglect center" is reported in polar coordinates to indicate its distance and direction from the page center. We apply our method to published examples of two-dimensional neglect. We find that neglect centers from different cancellation performances may not be statistically distinct even though they may occupy different quadrants. In addition, the net direction of neglect found by the coordinate method may differ from that inferred from measuring differences in quadrant omission totals. The suitability of the coordinate vs. the quadrant method will depend on the mechanism hypothesized for visuospatial exploration under particular test conditions. Using both approaches may detect different attentional biases operating during the same task. The coordinate method is appropriate for conventional cancellation testing. By incorporating the precise locations of all neglected stimuli and determining the net neglect direction in two dimensions, the technique may stimulate more comprehensive explanations for directional bias. PMID- 9339334 TI - "Wineglass" confabulations among brain-damaged alcoholics on the Wechsler Memory Scale-Revised visual reproduction subtest. AB - Confabulation is a clinically well-documented accompaniment of selective types of memory impairment, especially in brain-damaged alcoholics. This study reports specific occurrences of visual confabulation consisting of spontaneous alterations of Card D of the Visual Reproduction subtest of the Wechsler Memory Scale-Revised. The resemblance of a wineglass was fashioned by a 90-degree rotation into a "bowl and stem", observed in six of 30 brain-damaged alcoholics. There were no such instances in 132 other patients, including alcoholic controls, those with Parkinson's Disease, temporal lobe epileptics (pre- or post-surgery), and those with neurotoxic exposure. When asked, the subjects who identified the figure as a wineglass or similar drinking instrument reported that they had drawn it as originally shown to them. "Wineglass" confabulators had shorter periods of abstinence, longer drinking histories and lower intellectual functioning than their brain-damaged peers or an alcoholic control group. These findings lend support for the association of alcohol-related confabulation with visual, as well as previously-documented verbal material among brain-damaged alcoholics. PMID- 9339336 TI - Confabulation following rupture of posterior communicating artery. AB - In this study we report a patient, MG, who following rupture of left posterior communicating artery exhibited an amnesic-confabulatory syndrome. Neuropsychological examination showed severe impairment on episodic memory tasks, which were marred by florid but plausible and semantically appropriate confabulation. Performance on tasks involving various kinds of semantic knowledge was normal or only mildly impaired. Performance on tasks traditionally considered sensitive to frontal dysfunction was severely impaired with the exception of Cognitive Estimates where MG's performance was completely normal. There was no evidence of structural (CT scan) or metabolic (SPECT) damage to the frontal lobe. It is argued that tasks traditionally considered sensitive to frontal dysfunction are not specifically implemented by cognitive resources based on frontal structures. MG's confabulation is discussed in terms of a possible disruption of cognitive functions involved in the control of the subjective experience of feeling of remembering. PMID- 9339337 TI - Sniffing behaviour, or recognizing a lily by smell, but not recognizing a sock on sight. AB - We report a 65-year-old man with a post-anoxic encephalopathy who showed compulsive sniffing at available objects. This stereotyped environment-driven behaviour has not been previously described. Other compulsive environment-driven responses, such as manipulation and utilization of tools and hyperlexia, were also present. The disorder shared several features with the Kluver-Bucy syndrome where mouthing of objects, rather than smelling them, is common. The patient had a severe dementia, with amnesia, anomia, apraxia, and visual agnosia. Whereas he could not recognize very familiar objects on sight, he could in contrast correctly identify several familiar odours. Although sniffing was a compulsive and purposeless environment-driven behaviour, the question may be asked whether a relatively preserved olfactory recognition, in the presence of a severe disorder of visual recognition and knowledge, could have favoured a stereotyped exploration of objects by smelling. PMID- 9339338 TI - Factor analyses of handedness items in left and right-handed intellectually gifted and nongifted children. AB - Handedness questionnaire items from the General Scale of the Lateral Preference Schedule (Dean, 1988) were administered to 423 intellectually gifted and 226 nongifted children to investigate the underlying processes that contribute to laterality. Handedness items were factor analyzed using principal components (PC) analysis with varimax rotation. PC analyses computed separately for gifted and nongifted left and right-handed writers yielded very different factor structures. Left-handers (regardless of gifted status) tended to have factor structures marked by items that loaded saliently on more than one factor. Whereas a four factor solution best fit the data for the nongifted left-handers, a three factor solution was the best fit for the gifted left-handed children. The factor structure for the left-handed gifted children was marked by two factors where the item "draw" was the only item to load both positively and saliently, a pattern not evident among the nongifted left-handers. These results suggested different underlying patterns contributing to laterality among these groups of children. PMID- 9339339 TI - Right personal neglect following a left hemisphere stroke. A case report. AB - Neglect phenomena may occur in both extrapersonal and personal space. Whereas extrapersonal neglect has been found associated with both right- and left-sided brain lesions, no case of right personal neglect following a left-sided lesion has been so far reported. We describe a right-handed female patient who, after two left-hemisphere strokes, exhibited a florid personal neglect, but no extrapersonal neglect, anosognosia or somatoparaphrenia. The symptom persisted for a few weeks and then gradually disappeared. At least in the early phase of disease, a personal neglect can also be observed in patients with left brain damage. PMID- 9339340 TI - A novel nuclear gene, CBT1, essential for mitochondrial cytochrome b formation: terminal processing of mRNA and intron dependence. AB - We describe a new nuclear gene, CBT1 (Cytochrome B Termination), specifically involved in the generation of mature mRNA of cytochrome b in yeast mitochondria. Disruption of CBT1 (corresponding to ORF YKL 208W) results in a respiratory deficiency (no growth on acetate and ethanol, a reduced growth on glycerol, and a moderate growth on lactate). Cytochrome b is practically undetectable spectrally, while cytochromes a and a3 (cytochrome oxidase) appear unaffected by the disruption. Analysis of mitochondrial transcripts shows a reduced abundance of cytb mRNA, which in addition is approximately 200 nucleotides longer than that of the wild-type. Sequencing of the 3' region of the mutant cytb mRNA with an oligonucleotide primer positioned 148 nt downstream from the dodecamer sequence ("end-of-messenger" signal), demonstrates that the mutant transcript is extended beyond this position and is not processed at the conserved dodecamer cleavage site. The CBT1 gene product may be one of the components required for the exact 3' cleavage of the cytb messenger and may also be related to RNA splicing, since the intron-containing cytb gene is not as well expressed as the intron-less gene and the respiratory deficiency is more severe. We propose, that the CBT1 protein is necessary for the correct trimming of the end of cytb pre-mRNA and may be a part of the multi-component complex involved in this process. PMID- 9339341 TI - Association of transcripts from a group-I intron-containing gene with high sedimentation coefficient particles. AB - The mitochondrial gene coding for the large rRNA contains a self-splicing optional group-I intron (Sc-LSU.1) in some Saccharomyces cerevisiae strains. Although the mechanisms of splicing have been extensively studied, little is known about the possible interactions of this intron with other mitochondrial molecules such as proteins. Using glycerol gradients, we have compared the sedimentation coefficients of mitochondrial transcripts containing the Sc-LSU.1 intron in native yeast extracts and in purified RNA preparations. By comparing extracts from rho+ and rho- cells we have found that at least three RNA species containing the Sc-LSU.1 intron (4.5 kb, 2.7 kb and 1.2 kb respectively) are associated in vivo with a multimolecular complex of sedimentation coefficient 50S made up of nuclearly encoded proteins. Another RNA species of 2.7 kb, which may correspond to a cleavage at the dodecamer sequence of the intron, is not associated with the same particle. The possibility that the 50S particle corresponds to the mitochondrial ribosome or its precursor form(s) is discussed. PMID- 9339342 TI - The mcm17 mutation of yeast shows a size-dependent segregational defect of a mini chromosome. AB - Mini-chromosome-maintenance (mcm) mutants were described earlier as yeast mutants which could not stably maintain mini-chromosomes. Out of these, the ARS-specific class has been more extensively studied and is found to lose chromosomes and mini chromosomes due to a defect in the initiation of DNA replication at yeast ARSs. In the present study we have identified a number of mcm mutants which show size dependent loss of mini-chromosomes. When the size of the mini-chromosome was increased, from about 15 kb to about 60 kb, there was a dramatic increase in its mitotic stability in these mutants, but not in the ARS-specific class of mutants. One mutant, mcm17, belonging to the size-dependent class was further characterized. In this mutant, cells carried mini-chromosomes in significantly elevated copy numbers, suggesting a defect in segregation. This defect was largely suppressed in the 60-kb mini-chromosome. A non-centromeric plasmid, the TRP1ARS1 circle, was not affected in its maintenance. This mutant also displayed enhanced chromosome-III loss during mitosis over the wild-type strain, without elevating mitotic recombination. Cloning and sequencing of MCM17 has shown it to be the same as CHL4, a gene required for chromosome stability. This gene is non essential for growth, as its disruption or deletion from the chromosome did not affect the growth-rate of cells at 23 degrees C or 37 degrees C. This work suggests that centromere-directed segregation of a chromosome in yeast is strongly influenced by its length. PMID- 9339343 TI - Homologous recombination partly restores the secretion defect of underglycosylated acid phosphatase in yeast. AB - The majority of secreted acid phosphatase in Saccharomyces cerevisiae is encoded by the PH05 gene. The secretion level of this acid phosphatase is directly determined by its level of glycosylation. Consequently, PHO5-11-encoded acid phosphatase which lacks 11 of 12 glycosylation sites is only poorly secreted. We have isolated and characterized both UV- and EMS-induced variants, which are partly able to restore the secretion of acid phosphatase. Our data indicate that the improved secretion is caused by mitotic intrachromosomal recombination between the PHO5-11 allele and the homologous tandemly repeated PHO3 sequences, resulting in the restoration of glycosylation sites in PHO5-11. Two different recombination mechanisms, unequal sister-chromatid exchange and sister-chromatid gene conversion, are responsible for these alterations of the PHO5-11 locus. Thus, recombination between mutant and wild-type sequences are able to restore the ability of mutant yeast cells to secrete acid phosphatase. PMID- 9339344 TI - Development of pseudohyphae by embedded haploid and diploid yeast. AB - Diploid strains of S. cerevisiae are known to develop pseudohyphae in response to starvation for nitrogen. We report that both haploid and diploid yeast grow in a filamentous form when embedded in solid media. This is not a response to starvation, since yeast grown on rich media and overlaid with rich agar grow within the agar as pseudohyphae. While we find that the only element of diploidy required for formation of pseudohyphae in response to nitrogen starvation is the a1/alpha 2 repressor, pseudohyphal development by embedded cells does not require a1/alpha 2. Deletion of BUD 5 prevented the formation of pseudohyphae by embedded cells, suggesting that these structures are the result of ordered filament formation rather than agar penetration. Deletion of STE 12 prevented the formation of pseudohyphae by all cell types, showing that the same signal transduction pathway is used by embedded cells as by those responding to nitrogen starvation. Different cell types of yeast thus form filaments in response to several kinds of environmental stimuli. PMID- 9339345 TI - Variability of karyotypes and RAPD types in genetically related strains of Cryptococcus neoformans. AB - Variation in karyotypes and RAPD patterns of genetically related strains of Cryptococcus neoformans were analyzed. Capsular and filamentous mutants usually differ in their karyotypes from wild-types, but the RAPD patterns were found to be similar. Karyotype differences were observed in most heterothallic matings, but RAPD patterns remained identical. After self-sporulation of a diploid strain, minor chromosomal length polymorphism and minor changes in the RAPD types occurred. Three mechanisms, either alone or in combination, may in varying degrees contribute to the karyotype variation of C. neoformans: (1) mitotically induced changes; (2) karyotype changes as a result of meiotic recombination, and (3) mutagen-induced changes. The present data do not support the meiotic maintenance hypothesis, which claims that the amount of CLP generated is inversely proportional to the frequency of meiosis. PMID- 9339346 TI - Heterokaryon incompatibility blocks virus transfer among natural isolates of black Aspergilli. AB - The extent of heterokaryon (also termed somatic or vegetative) incompatibility among black Aspergillus strains was examined using nitrate non-utilising mutants selected on chlorate medium. Pairings of complementary mutants showed that somatic compatibility between different strains is exceptional in natural populations of the asexual black Aspergilli. Mycoviruses are present in a considerable fraction of the sampled natural population, but surprisingly, horizontal transfer of mycoviruses only occurs-at least under laboratory conditions-between the (very rare) compatible combinations of strains. Thus, unlike other fungal species, somatic incompatibility in black Aspergilli efficiently blocks virus transfer. Viruses present in black Aspergillus isolates are highly efficiently transmitted to asexual progeny. PMID- 9339347 TI - Dominant mutations affecting both sporulation and sterigmatocystin biosynthesis in Aspergillus nidulans. AB - The initiation of conidiophore development in the filamentous fungus Aspergillus nidulans is a complex process requiring the activities of several genes including fluG, flbA, flbB, flbC, flbD, and flbE. Recessive mutations in any one of these genes result in greatly reduced expression of the brlA developmental regulatory gene and a colony morphology described as fluffy. These fluffy mutants have somewhat diverse phenotypes but generally grow as undifferentiated masses of vegetative hyphae to form large cotton-like colonies. In this paper we describe a genetic screen to identify dominant mutations resulting in similar fluffy colony morphologies. We have identified 36 dominant fluffy mutant strains and shown that 29 of these mutants have greatly reduced brlA expression as compared to wild type. In addition, we have found that 19 of these mutants are not only developmentally altered but also fail to produce the toxic, carcinogenic, secondary metabolite sterigmatocystin. At least three of the mutants isolated result from dominant activating mutations in fadA which encodes the G alpha subunit of a heterotrimeric G-protein. Another of the mutants results from a dominant interfering mutation in brlA. We discuss the approaches taken to characterize these potentially important regulators of growth, development and secondary metabolism. PMID- 9339348 TI - Hydrophobin gene srh1, expressed during sporulation of the biocontrol agent Trichoderma harzianum. AB - A cDNA clone encoding a spore-related hydrophobin, SRHI, was isolated from a cDNA bank prepared from mRNA induced in sporulating cultures of Trichoderma harzianum by heterologous hybridization using the hfb2 gene encoding a spore-bound hydrophobin of Trichoderma reesei as a probe. Based on sequence similarity the predicted protein was identified as a new member of the class-II hydrophobin family. Including the signal sequences, SRHI has 65% and 56% amino-acid similarity with the T. reesei hydrophobins HFBII and HFBI, respectively, being less similar with other hydrophobins. srh1 is present as one copy in the T. harzianum genome. It is highly expressed under sporulating conditions, both in submerged as well as in aerial cultures. Moreover, nutrient limitation induces srh1 expression. PMID- 9339349 TI - A temperature-sensitive mutation of Coprinus cinereus, hyt1-1, that causes swelling of hyphal tips. AB - The TU25 mutant strain of the basidiomycete Coprinus cinereus grows well at 28 degrees C but not at 37 degrees C. Microscopic examination revealed that TU25 exhibited swelling at hyphal apices after a shift-up from 28 degrees C to 37 degrees C. The temperature sensitivity and hyphal swelling co-segregated through meiosis as expected for a single Mendelian factor, designated hyt1 (hyphal tip). Both defects could be suppressed by the presence of osmotic stabilizers in the medium, suggesting that the hyt1 gene product is required for proper cell-wall function. Chemical analysis of the cell walls, however, failed to detect any clear difference in the composition of the cell-wall polysaccharides between the wild-type and TU25. A DNA fragment which complements the hyt1-1 mutation was cloned and sequenced. The predicted protein in the ORF essential for complementation of hyt1-1 is a novel protein. PMID- 9339350 TI - Sequence variation of the rDNA ITS regions within and between anastomosis groups in Rhizoctonia solani. AB - Sequence analysis of the rDNA region containing the internal transcribed spacer (ITS) regions and the 5.8s rDNA coding sequence was used to evaluate the genetic diversity of 45 isolates within and between anastomosis groups (AGs) in Rhizoctonia solani. The 5.8s rDNA sequence was completely conserved across all the AGs examined, whereas the ITS rDNA sequence was found to be highly variable among isolates. The sequence homology in the ITS regions was above 96% for isolates of the same subgroup, 66-100% for isolates of different subgroups within an AG, and 55-96% for isolates of different AGs. In neighbor-joining trees based on distances derived from ITS-5.8s rDNA sequences, subgroups IA, IB and IC within AG-1 and subgroups HG-I and HG-II within AG-4 were placed on statistically significant branches as assessed by bootstrap analysis. These results suggest that sequence analysis of ITS rDNA regions of R. solani may be a valuable tool for identifying AG subgroups of biological significance. PMID- 9339351 TI - Identification of the ure1+ gene encoding urease in fission yeast. AB - Cloning and sequencing of the ure1+ gene of Schizosaccharomyces pombe indicated that it encodes the urease which had been biochemically identified. The fission yeast urease has a one-subunit structure like those from plants but different from bacterial ureases which are composed of two or three distinct subunits. Genetic analyses showed that the ure1+ gene product is actually involved in urea metabolism. PMID- 9339352 TI - Diagnosis and treatment of diseases of the aorta. PMID- 9339353 TI - A psychobiologic approach to pediatric pain: Part II. Prevention and treatment. PMID- 9339354 TI - A gene in human chromosome band Xq28 (GABRE) defines a putative new subunit class of the GABAA neurotransmitter receptor. AB - We have isolated and sequenced a novel human gene (GABRE) of the GABAA neurotransmitter receptor family. A cDNA sequence of the gene coding for a 506 amino acid protein was identified, representing a member of a putative new class (epsilon) of the GABAA receptor. The gene is transcribed at least at low level in several different tissues, with the highest levels being detected in adult heart and placenta. Alternative splicing of GABRE transcripts isolated from different tissues was observed at multiple positions of the gene, yielding an unusually complex variety of cDNA variants. The structure of the 5' region of most cDNAs is compatible with expression of protein sequence epsilon only in adult brain, whereas in other tissues, the majority of transcripts code for truncated protein sequences. The GABRE gene extends over 14 kb and is clustered together with the alpha 3 and the putative beta 4 GABAA receptor subunit genes in an approximately 0.8-Mb interval in chromosome band Xq28, located in the candidate regions of two different neurologic diseases. Based on features of conservation of protein sequences, gene structure, and genomic organization of GABAA receptor gene clusters, we propose that the epsilon and gamma subunit genes have a common ancestor and that GABAA receptor gene clusters in the human genome have diverged by multiple duplication events of an ancestral gene cluster containing one each alpha, beta, and gamma/epsilon precursor gene. PMID- 9339355 TI - A 1.2-megabase BAC/PAC contig spanning the 14q13 breakpoint of t(2; 14) in a mirror-image polydactyly patient. AB - We previously assigned a 14q13 breakpoint of t(2; 14) in a patient with mirror image polydactyly to a segment between two loci, AFM200ZH4 and D14S306, within a genetic distance of 0.6 cM. In the present study, we constructed a 1.2-Mb high resolution physical map with a contig composed of 16 bacterial artificial chromosomes (BACs) and 6 P1-derived artificial chromosomes (PACs) at a region around the breakpoint, extending from D14S75 to D14S728 loci. Thirty-four novel sequence-tagged sites (STSs) were also characterized at this region. Of nine ESTs that had been mapped between D14S75 and D14S288, T99065 was confirmed to be in two BAC clones, B102 and B319. This BAC/PAC contig with STSs is useful for further genomic sequencing, for construction of a transcription map, and for the isolation of the putative gene for mirror-image polydactyly. PMID- 9339356 TI - Molecular characterization and pattern of tissue expression of the gene for neutrophil gelatinase-associated lipocalin from humans. AB - Neutrophil gelatinase-associated lipocalin (NGAL) is a 25-kDa lipocalin first identified as a protein stored in specific granules of the human neutrophil. The protein is believed to bind small lipophilic substances such as bacterial derived formylpeptides and lipopolysaccharides (LPS) and might function as a modulator of inflammation. To characterize the regulation of NGAL further, we have cloned and sequenced a 5869-bp region of the NGAL gene including 1695 bp of the 5' nontranscribed region and a 3696-bp coding region encompassing seven exons and six introns. The transcriptional start sites were identified by an RNase protection assay. The NGAL gene is highly homologous to the mouse gene 24p3. NGAL was expressed in bone marrow and in tissues that are prone to exposure to microorganisms. Potential cis-acting elements were identified in the promoter region of the NGAL gene by computer analysis and include binding sites for CTF/CBP, the hematopoietic transcription factors GATA-1 and PU.1, and the LPS inducible factor NF-kappa B. PMID- 9339357 TI - Mapping of replication origins and termination sites in the Duchenne muscular dystrophy gene. AB - The replication structure of the human dystrophin gene in cultured masculine erythroleukemia cells (line HEL 92.1.7) was studied using the replication direction assay. This gene is organized into at least six replicons ranging in size from 170 to more than 500 kb. One of the replicon junctions (sites of replication termination) was mapped to intron 44, i.e., roughly in the same area where the major recombination hot spot is located. A replicon junction was also found between the muscle and the brain promoters. The two replicons mapped in the present study are highly asymmetric, as the distances covered by the replication forks moving in opposite directions from the same origin differ by more than threefold. PMID- 9339358 TI - Cloning of mouse type XV collagen sequences and mapping of the corresponding gene to 4B1-3. Comparison of mouse and human alpha 1 (XV) collagen sequences indicates divergence in the number of small collagenous domains. AB - We report on full-length mouse type XV collagen cDNAs that encode a 1367-residue alpha 1(XV) chain. The amino acid sequences of the mouse and previously characterized human alpha 1(XV) chains exhibit an overall identity of 72%. The highest homology between these chains and to the structurally related type XVIII collagen is observed in their C-terminal noncollagenous domains. Although the mouse and human alpha 1(XV) chains are highly homologous and similar in their overall domain structure, the mouse chain contains only seven collagenous domains, whereas the human chain contains nine. Northern analysis of several mouse tissues indicated strong hybridization in the case of heart and skeletal muscle RNAs and moderate signals with kidney, lung, and testis RNAs. Analysis of type XV collagen mRNA levels at different stages of mouse embryonic development indicated a marked increase in the level between 11 and 15 days of development, which coincides with pronounced development of the muscles, heart, and vascular system in the mouse embryo. The mouse gene for type XV collagen was mapped by fluorescence in situ hybridization to chromosome 4, band B1-3. This result indicates that the mouse type XV collagen gene and its human counterpart are located in the chromosomal segments with conserved syntenies. PMID- 9339359 TI - FISH probes for mouse chromosome identification. AB - P1 clones near the telomeres and centromeres of each mouse chromosome except Y have been selected from a mouse genomic library and mapped using fluorescence in situ hybridization (FISH). Each clone was selected to contain a genetically mapped polymorphic DNA sequence as close as possible to the centromere or telomere of a chromosome. The genetic distance from the various P1 clones to the most distal genetically mapped polymorphic sequence ranged from 0 for about half of the clones to 6.7 cM for the probe at the telomere of chromosome 14. The average distance to the most distal or proximal chromosome marker was 1.5 cM. The use of FISH with these probes for mouse chromosome identification during comparative genomic hybridization is illustrated. PMID- 9339360 TI - Analysis of exon/intron structure and 400 kb of genomic sequence surrounding the 5'-promoter and 3'-terminal ends of the human glypican 3 (GPC3) gene. AB - GPC3, the gene modified in the Simpson-Golabi-Behmel gigantism/overgrowth syndrome (SGBS), is shown to span more than 500 kb of genomic sequence, with the transcript beginning 197 bp 5' of the translational start site. The Xq26.1 region containing GPC3 as the only known gene has been extended to > 900 kb by sequence analysis of flanking BAC clones. Two GC isochores (40.6 and 42.6% GC) are observed at the 5' and 3' ends of the locus, with a large repertoire of repetitive sequences that includes an unusual cluster of four L1 elements > 92% identical over 2.8 kb. Eight exons, accounting for the full 2.4-kb GPC3 cDNA, have been sequenced along with neighboring intronic regions. PCR assays have been developed to amplify each exon and exon/intron junction sequence, to help discriminate instances of SGBS among individuals with overgrowth syndromes and to facilitate mutational analysis of lesions in the gene. PMID- 9339361 TI - Cosmid contig and transcriptional map of three regions of human chromosome 21q22: identification of 37 novel transcripts by direct selection. AB - Human chromosome 21 is associated with many disorders, including Down syndrome (DS). In an effort to identify genes involved in brain development or function and therefore implicated in the mental retardation associated with DS, we chose YACs from three regions of chromosome 21: a region within the so-called "Down syndrome critical region," a region proximal to it, and one distal to it. We made cosmid libraries from these YACs and generated high-resolution physical maps by constructing cosmid contigs. These are the first cosmid contigs on chromosome 21 outside the critical region. The cosmids were used for direct selection of cDNAs to isolate chromosome 21 expressed sequences. We have isolated 45 nonredundant partial cDNAs and mapped these back to the cosmid contigs. We isolated 3 nonoverlapping portions of DSCR1 and a part of GIRK2 and identified 3 nonoverlapping partial cDNAs with similarity to the rat Dyrk gene, which turned out to be the human homologue (MNB) of the Drosophila minibrain gene. Twelve sequences had matches with either STS or EST entries in the databases, including a chromosome 21 EST, a chromosome 21 STS, and 6 unmapped expressed sequence entries. Only 1 sequence resulted in a match with a protein entry. The remaining 25 sequences revealed no similarity to any database entry. All of these partial cDNAs are expressed as determined by Northern blotting or by RT-PCR. PMID- 9339362 TI - Cloning of GPR37, a gene located on chromosome 7 encoding a putative G-protein coupled peptide receptor, from a human frontal brain EST library. AB - A cDNA sequence encoding a putative peptide-specific G-protein-coupled receptor (GPR37) was isolated from a set of human brain frontal lobe expressed sequence tags. The GPR37 cDNA predicts a single open reading frame coding for a 613-amino acid protein with seven hydrophobic transmembrane domains. The GPR37 genomic sequence was mapped to chromosome 7q31, and it was isolated upon screening of a chromosome 7-specific genomic library. The GPR37 gene spans more than 25 kb and contains two exons and a single intron which interrupts the GPR37 cDNA within the sequence encoding the presumed third transmembrane domain. Northern blot analysis with GPR37 probes revealed a main 3.8-kb mRNA and a less abundant 8-kb mRNA, both expressed in human brain tissues, particularly in corpus callosum, medulla, putamen, and caudate nucleus. The lowest level of expression was detected in cerebellum. The 3.8-kb mRNA is also less abundantly expressed in liver and placenta. Although the ligand for the putative GPR37 receptor has not been identified, its deduced amino acid sequence shows a high degree of homology (approximately 40% in the transmembrane regions) with most mammalian peptide specific G-protein-coupled receptors and particularly with the human endothelin B, bombesin-BB1, and bombesin-BB2 receptors. PMID- 9339363 TI - Estimation of the age of the ancestral arginine3500-->glutamine mutation in human apoB-100. AB - Familial defective apoB-100 (R3500Q) [FDB (R3500Q)] is caused by a mutation in the apoB gene (2p23.24). Almost all individuals with this disorder are of European descent, and in almost all cases the mutation is on a chromosome with a rare haplotype (194) at the apoB locus, suggesting that all FDB (R3500Q) probands are descended from a common ancestor in whom the original mutation occurred. The distribution of the mutation is consistent with an origin in Europe 6000-7000 years ago. We have estimated the amount of recombination between the apoB gene and markers on chromosome 2 in 34 FDB (R3500Q) probands in whom the mutation is on a 194 haplotype. Significant linkage disequilibrium was found between the apoB gene and marker D2S220. We have identified three YACs that contain the apoB gene and D2S220. The shortest restriction fragment common to the three YACs that contained both loci was 240 kb long. No shorter fragments with both loci were identified. On the assumption that 1000 kb corresponds to 1 cM, we deduce that the recombination distance between D2S220 and the apoB gene is about 0.24 cM. Combining this value with the linkage disequilibrium observed between the two loci in the probands, we estimate that the ancestral mutation occurred about 270 generations ago. We postulate that the original mutation occurred in the common ancestor of living FDB (R3500Q) probands, who lived in Europe about 6750 years ago. The errors in this estimate are discussed. PMID- 9339364 TI - Novel genes mapping to the critical region of the 5q- syndrome. AB - The 5q- syndrome is a myelodysplastic syndrome with specific hematological features and a good prognosis. Using molecular mapping techniques, we have previously defined the critical region of gene loss of the 5q- chromosome in the 5q- syndrome as the approximately 5-Mb region at 5q31-q33 flanked by the genes for FGF1 and IL12B. This region is completely represented by a series of overlapping YACs, and we are currently generating a transcription map with the aim of identifying the tumor-suppressor gene associated with the development of the 5q- syndrome. In this study two techniques have been used: first, the screening of full-length cDNA libraries with radiolabeled YACs and second, the mapping of chromosome 5-specific expressed sequence tags (ESTs) to a YAC contig. A 1-Mb YAC contig encompassing the CSF1R gene has been used to screen a fetal brain cDNA library, and this has resulted in the identification of two genes comprising one known gene previously localized to the region (ADRB2) and one known gene previously unlocalized. Six of 135 chromosome 5-specific ESTs were localized by PCR screening to the YAC contig mapping to the critical region of the 5q- syndrome. IMAGE cDNA clones for each of the six ESTs have been obtained. These seven (excluding ADRB2) newly assigned cDNA clones were subjected to further analysis. The expression patterns of each of the cDNA clones have been established in a range of human tissues, including bone marrow. Six of seven cDNAs are expressed in human bone marrow. Six of seven cDNAs have no known homology to any deposited human sequences, and one (C29) is dihydropyrimidinase related protein-3, a member of a novel gene family. Genomic localization and expression patterns would suggest that these newly assigned cDNAs represent potential candidate genes for the 5q- syndrome. PMID- 9339365 TI - Celsr1, a neural-specific gene encoding an unusual seven-pass transmembrane receptor, maps to mouse chromosome 15 and human chromosome 22qter. AB - We have identified Celsr1, a gene that encodes a developmentally regulated vertebrate seven-pass transmembrane protein. The extracellular domain of Celsr1 contains two regions each with homology to distinct classes of well-characterized motifs found in the extra-cellular domains of many cell surface molecules. The most N-terminal region contains a block of contiguous cadherin repeats, and C terminal to this is a region containing seven epidermal growth factor-like repeats interrupted by two laminin A G-type repeats. Celsr1 is unique in that it contains this combination of repeats coupled to a seven-pass transmembrane domain. As part of the characterization of the Celsr1 gene, we have determined its chromosomal map location in both mouse and human. The European Collaborative Interspecific Backcross (EUCIB) and BXD recombinant inbred strains were used for mapping Celsr1 cDNA clones in the mouse, and fluorescence in situ hybridization was used to map human Celsr1 cosmid clones on metaphase chromosomes. We report that Celsr1 maps to proximal mouse Chromosome 15 and human chromosome 22qter, a region of conserved synteny. Reverse transcriptase-polymerase chain reaction analysis and in situ hybridization were used to determine the spatial restriction of Celsr1 transcripts in adult and embryonic mice. The results presented here extend our previous finding of expression of the Celsr1 receptor in the embryo and show that expression continues into adult life when expression in the brain is localized principally in the ependymal cell layer, choroid plexus, and the area postrema. PMID- 9339366 TI - Characterization of the gene (VBP1) and transcript for the von Hippel-Lindau binding protein and isolation of the highly conserved murine homologue. AB - A single-copy, widely expressed gene of at least 30 kb and six exons was discovered via a hybrid mRNA resulting from an inversion causing hemophilia A. A segment of the 1.7-kb message of this gene has been shown by others to encode a protein (named VBP1) interacting with the product of the von Hippel-Lindau gene and thus is expected to participate in pathways involving this tumor suppressor gene. The mouse VBP1 message we isolated encodes a polypeptide of 160 residues absolutely identical to that of human. Even the 3' untranslated tails of the mRNAs show 68% conservation, and both use the unusual ATTAAA polyadenylation signal. The mouse gene has a single transcription start while the human homologue has two major starts and a minor start. This could result in amino-end extensions of the human protein. A polymorphism with 42% heterozygosity in the United Kingdom population was detected in the 3' tail of the message. VBP1 is unlike other known proteins but a consensus for tyrosine phosphorylation possibly suggests regulation by kinases. PMID- 9339368 TI - Identification, partial characterization, and genetic mapping of kinesin-like protein genes in mouse. AB - Microtubule-dependent motors of the kinesin superfamily have undergone structural and functional diversification during evolution and play crucial roles in cell division and intracellular transport. Degenerate oligonucleotides homologous to highly conserved regions of sequence within the motor domain were used in a polymerase chain reaction to isolate five new members (KIF3C, KIFC2, KIFC3, KIFC4, and KIF22) of the kinesin superfamily from a mouse brain cDNA library. Northern analysis showed that KIF3C and KIFC2 are expressed mainly in neural tissues, that KIFC4 and KIF22 are expressed primarily in proliferative tissues and cell lines, and that KIFC3 is apparently ubiquitous. To elucidate the organization of genes encoding kinesin-like motors in the mouse genome and to explore the potential associations of these genes with classical mouse mutations or human genetic diseases, these new genes as well as genes encoding the previously reported KIF3A and KIF3B motors were mapped to mouse chromosomes by using an interspecific backcross panel of DNAs from The Jackson Laboratory. The data indicate that the gene KIFC4 is present in three copies in the mouse genome on chromosomes 13 (KIFC4A), 10 (KIFC4B), and 17 (KIFC4C). The gene KIF22 is present in two copies on chromosomes 7 (KIF22A) and 1 (KIF22B). The genes KIF3A, KIF3B, KIF3C, KIFC2, and KIFC3 are each single loci and map to chromosomes 11, 2, 12, 15, and 8, respectively. PMID- 9339367 TI - Genomic structure and chromosomal localization of a human myo-inositol monophosphatase gene (IMPA). AB - Manic-depressive illness is a serious psychiatric disorder that in many, but far from all, patients can be treated with lithium. The main causes for discontinuation of lithium therapy are unpleasant or serious side effects and lack of response. The reason for the striking variation in clinical efficacy of lithium treatment among bipolar patients is not known. The enzyme myo-inositol monophosphatase (IMPase) has been postulated as a target for the mood-stabilizing effects of lithium, but variation in the coding region of the human IMPA gene encoding IMPase activity has not been observed in manic-depressive patients (Steen et al., Pharmacogenetics, 1996, 6, 113-116). It is nevertheless conceivable that polymorphisms or mutations in the noncoding regions of this gene could influence the lithium response in psychiatric patients. As a first step in investigating this possibility, we here report the genomic structure of the human IMPA gene. The gene is composed of at least nine exons and covers more than 20 kb of sequence on chromosome 8q21.13-q21.3. In the 3'-untranslated part of the gene, we observed a polymorphism (a G to A transition) and also two short sequences similar to the inositol/cholin-responsive element consensus. Finally, we postulate that two additional IMPA-like transcripts originate from the human genome, one from a position close to IMPA itself on chromosome 8 and the other from chromosome 18p. Our data may contribute to the identification of genetic factors involved in the pathogenesis and determination of treatment response in manic-depressive illness. PMID- 9339369 TI - Dp140: alternatively spliced isoforms in brain and kidney. AB - Dp140, a protein composed of the distal rod domain and carboxy-terminal domain of dystrophin, is expressed only in brain and kidney; transcription is initiated at a unique first exon located in dystrophin intron 44. Both tissues express specific isoforms with distinct alternative splicing of exons 71-74 and 78. The carboxy-terminal domain of Dp140 is identical to that of full-length 427-kDa dystrophin, and it is this region that is associated, with the "dystrophin associated protein" complex in skeletal muscle. Alternative splicing encodes domains that have been shown to be protein-binding regions; thus this alternative splicing may regulate the association of Dp140 with integral membrane proteins. The 5' flanking region of genomic DNA adjacent to the Dp140 first exon contains a variety of transcription factor-binding motifs, some of which could regulate neuroglial-specific gene expression. PMID- 9339370 TI - Mapping of 228 ESTs and 26 genes into an integrated physical and genetic map of human chromosome 17. AB - We have integrated genetic and physical mapping data for chromosome 17 subdivided into 26 bins, by using a panel of chromosome 17 deletion somatic cell hybrids. One hundred four short tandem repeat and STS markers have been localized into these bins and have enabled the ordering of 288 ESTs and 26 genes, including 142 ESTs that had not been previously sublocalized on chromosome 17. The mapping information of several genetic maps, as well as information obtained by radiation hybrid and STS content mapping of YACs, has been integrated using this hybrid panel. Although existing mapping information for chromosome 17 was generally consistent for many ESTs previously mapped, the map presented here further refines the location of ESTs, as well as demonstrating a number of discrepancies found in the 17q24-q25 region. We attribute these discrepancies to the fact that the current radiation hybrid panels were selected for retention of the thymidine kinase gene at 17q25, as well as to a low concentration of YAC contigs in this region. These data illustrate the benefit of combining multiple mapping techniques to obtain the greatest accuracy. The integration of maps developed by different methods will generate the most accurate genome maps, which may then be used for the generation of large insert clone contigs for chromosome sequencing. Additionally, accurate transcript maps generated by ESTs will greatly speed the isolation of genes linked to disease loci. PMID- 9339371 TI - Construction of a 3-Mb contig and partial transcript map of the central region of mouse chromosome 11. AB - We report the establishment of a high-resolution genetic map, a physical map, and a partial transcript map of the Ames dwarf critical region on mouse chromosome 11. A contig of 24 YACs and 13 P1 clones has been assembled and spans approximately 3 Mb from Flt4 to Tcf7. A library of approximately 1000 putative transcript clones from the region was prepared using exon amplification and pituitary cDNA selection. Ten novel transcripts were partially characterized, including a member of the olfactory receptor family, an alpha-tubulin-related sequence, and a novel member of the cdc2/CDC28-like kinase family, Clk4. The location of Prop1, the gene responsible for Ames dwarfism, has been localized within the contig. This contig spans a region of mouse chromosome 11 that exhibits linkage conservation with human chromosome 5q23-q35. The strength of the genetic map and genomic resources for this region suggest that comparative DNA sequencing of this region could reveal the genes responsible for other mouse mutants and human genetic diseases. PMID- 9339372 TI - Regions of human chromosome 2 (2q32-q35) and mouse chromosome 1 show synteny with the pufferfish genome (Fugu rubripes). AB - We have isolated and sequenced a cosmid clone from the compact genome of the Japanese pufferfish (Fugu rubripes) containing portions of three genes that have the same order as in human. The gene order is microtubule-associated protein (MAP 2), myosin light chain (MYL-1), and carbamoyl phosphate synthetase (CPS III). The intron-exon organization of Fugu CPS III is identical with that of rat CPS I, although the equivalent genomic fragments of rat and Fugu CPS span 87.9 and 21 kb, respectively. This is the first report of a piscine CPS III genomic structure and predicts a close evolutionary link between CPS III and CPS I. The 8-kb intergenic region between MYL-1 and CPS gave no clear areas of transcription factor-binding sites by pairwise comparison with shark or rat CPS promoter regions. However, there was a match with the rat myosin light chain 2 (MLC-2) gene promoter and a MyoD transcription factor-binding site 874 bp upstream of the MYL-1 gene. PMID- 9339373 TI - The genomic organization and the full coding region of the human PAX7 gene. AB - The PAX7 gene encodes a transcription factor that is a member of the PAX family of developmental control genes. In addition to playing a role in embryogenesis, PAX genes appear to be of importance in a number of diverse human diseases and cancers. The PAX7 gene maps to human chromosomal region 1p36 and therefore is a potential candidate for human disorders linked to this region. In particular, a rearrangement of the PAX7 gene by chromosomal translocation is frequently found in alveolar rhabdomyosarcoma tumors. Here, we cloned a cDNA containing the full coding region of the human PAX7 gene and determined its genomic organization. The gene encodes a predicted protein of 520 amino acids that is 47 amino acids longer at the carboxy end than the highly related PAX3 protein. The coding region of the gene is interrupted by seven introns, the positions and lengths of which are similar to those of the corresponding introns of the PAX3 gene. Sequence data for exon/intron boundaries of PAX7 exons 1, 5, 6, 7, and 8 were determined and, together with previously published data for exons 2, 3, and 4, provide the complete sequence information for mutation analysis of the human PAX7 gene. PMID- 9339374 TI - A new subfamily of vertebrate calpains lacking a calmodulin-like domain: implications for calpain regulation and evolution. AB - Calpains are calcium-dependent intracellular nonlysosomal proteases that are believed to participate in signal transduction. In vertebrates, five different calpains have so far been identified, of which three, mu-, m-, and mu/m-calpain, are ubiquitously expressed while the other two, nCL-1 (p94) and nCL-2, exhibit a restricted tissue distribution. We have identified two new vertebrate calpain genes, Capn5 and Capn6. The human and mouse amino acid sequences of these new calpains are the most divergent of the vertebrate calpains identified. They possess most of the residues conserved in calpain family members but the C terminal region lacks any homology to the calmodulin-like domain of other vertebrate calpains. They both exhibit significant homology over the entire coding region to the protein encoded by the gene tra-3, involved in nematode sex determination, and Capn5 may represent its vertebrate orthologue. The predicted Capn6 protein lacks critical active site residues and may not be proteolytically active. Both genes are differentially expressed in human tissues with highest RNA levels for Capn5 occurring in the testis, liver, trachea, colon, and kidney, while Capn6 is highly expressed only in the placenta sample of the 50 tissues examined. Phylogenetic analysis suggests that the vertebrate calpains arose through a series of gene duplication events that began before the initial divergence of the vertebrate and invertebrate lineages. The discovery of these two new calpains highlights a hitherto unknown complexity of the calpain family with subclasses perhaps possessing different modes of regulation. PMID- 9339375 TI - Variable expression of hepatic glucokinase in mice is due to a regulational locus that cosegregates with the glucokinase gene. AB - The Gk activity locus affects expression of hepatic glucokinase (GK) in mice. Analysis of microsatellites within the mouse GK gene locus revealed two major haplotypes in 19 of 22 inbred strains predictive of either high or low hepatic GK gene expression. C3H/HeJ mice, a high-activity strain, and two other wild-derived strains contain less common haplotypes. No coding sequence differences were found in hepatic GK-coding sequences from representative high and low Gk activity strains, thereby excluding kinetic abnormalities as the basis for hepatic GK activity differences. Screening of approximately 10 kb of potential regulatory DNA, including all eight known and three of four newly identified DNase I hypersensitive sites, by restriction enzyme fingerprinting-single-strand conformation analysis revealed a tetranucleotide microsatellite, the length of which was also predictive of the Gk activity phenotype. This tetranucleotide repeat is located in the first intron of the hepatic transcription unit and lies close to a newly identified liver-specific DNase I-hypersensitive site. These results indicate that the Gk activity alleles are a regulational locus associated with the GK gene locus. PMID- 9339376 TI - The murine DNA-PKcs gene consists of 86 exons dispersed in more than 250 kb. AB - The DNA-PKcs gene encodes the 465-kDa catalytic subunit of DNA-dependent protein kinase (DNA-PK), which associates with heterodimeric autoantigens Ku70 and Ku80 and exhibits protein kinase activity depending on DNA double-strand breaks. The gene is also responsible for the aberration in severe combined immune deficiency (SCID) mice, which exhibit a high sensitivity to ionizing radiation and abnormal DNA rearrangement of immunoglobulin and T cell receptor genes. There is further evidence that DNA-PKcs phosphorylates various proteins involved in DNA replication, transcription, repair, and recombination. Nevertheless the structure/function relationship in this huge molecule is virtually unknown. We determined the exons and introns of the murine DNA-PKcs gene by the long-distance polymerase chain reaction method. The murine DNA-PKcs gene consists of 86 exons distributed in a region of more than 250 kb. The average size of the exons is 140 bp. All the splicing sites conform to the GT/AG rule. The SCID mutation site (Tyr4046) has been identified in exon 85. The genomic structure of the DNA-PKcs gene provides clues for the study of various functional domains in this macromolecule. PMID- 9339377 TI - Genomic organization and molecular characterization of a gene encoding HsPXF, a human peroxisomal farnesylated protein. AB - A protein modification essential for the cellular sorting of many biologically relevant proteins is the covalent attachment of prenyl lipids by specific transferases. Isoprenylation is known to render protein domains hydrophobic, thereby facilitating the interaction with lipid bilayers and/or membrane proteins. The target for the modification with farnesyl groups is the COOH terminal sequence CaaX. Among the variety of farnesylated proteins the only one reported so far to be located to peroxisomes is the 37-kDa peroxisomal farnesylated hamster protein PxF. Recently we published data on the cDNA of the human gene HK33 (A. Braun et al., 1994, Gene 146: 291-295), which was revealed to be the human ortholog of PxF and was consequently renamed HsPXF. The genomic structure, molecular characterization, and evolutionary conservation of HsPXF are described herein. The exact location of the gene was defined as chromosome 1q22. The gene spans a region of approximately 9 kb, containing eight exons and seven introns. The 5' upstream region showed two potential Sp1-binding sites and an Alu repetitive sequence. Luciferase reporter activating capacity confirmed the presumed promoter activity of this region. On the transcriptional level, we detected four splice variants originating either from exon skipping or from alternative splicing events. For the HsPXF protein, a carboxyterminal farnesylation at cysteine residues was demonstrated. Through the use of HsPXF specific antibodies, the protein was shown to be attached to the outer surface of peroxisomes. This localization together with the similarity to a peroxisomal assembly protein from Saccharomyces cerevisiae suggests HsPXF is involved in the process of peroxisomal biogenesis or assembly. PMID- 9339378 TI - Genomic organization and functional analysis of the murine protein phosphatase 1c gamma (Ppp1cc) gene. AB - Protein phosphatase 1 holoenzymes are composed of catalytic subunits in combination with various regulatory subunits. In rodents, four different catalytic isoforms are known, PP1c alpha, -delta, -gamma 1, and -gamma 2. Here we describe the genomic organization of the murine Ppp1cc gene that encodes the PP1c gamma 1 and PP1c gamma 2 isoforms. We determined that Ppp1cc maps to F1.2-G1.2 on chromosome 5 by FISH mapping. Southern hybridization and analysis of cross hybridizing genomic clones revealed four Ppp1cc-related pseudogenes in the mouse genome. The authentic Ppp1cc gene encodes two isoforms, PP1c gamma 1 and PP1c gamma 2, that arise from alternative splicing and differ by retention of the last intron. The introns of Ppp1cc are flanked by short direct repeats, the significance of which is not clear. Both isoforms retain phosphatase function since they are able to complement the cold-sensitive PP1 defect caused by the dis2-11 mutation in the fission yeast Schizosaccharomyces pombe. PMID- 9339379 TI - Differential expression of XAP5, a candidate disease gene. AB - We have isolated a full-length cDNA corresponding to the XAP5 gene in Xq28. An unusual feature of the cDNA is that it contains runs of CCG repeats in the 5' untranslated region, typical of genes that exhibit anticipation. It has a striking pattern of differential expression and is greatly enhanced in various fetal tissues. This predicted protein encodes a unique 339-amino-acid polypeptide that contains a large percentage of highly charged residues and a possible nuclear localization signal. A comparison to genomic sequence shows that XAP-5 comprises 13 exons spanning 6.5 kb. An examination of the human population indicates that the longest CCG run is polymorphic and varies in length from 8 to 12 repeats. PMID- 9339380 TI - Molecular cloning, cDNA sequence analysis, and chromosomal localization of mouse Pkd2. AB - The gene responsible for the second form of autosomal dominant polycystic kidney disease, PKD2, has recently been identified. We now describe the cloning, genomic localization, cDNA sequence, and expression analysis of its murine homologue, Pkd2. The cloned cDNA sequence is 5134 bp long and is predicted to encode a 966 amino-acid integral membrane protein with six membrane-spanning domains and intracellular NH2 and COOH termini. Pkd2 is highly conserved with 91% identity and 98% similarity to polycystin-2 at the amino acid level. Pkd2 mRNA is widely expressed in mouse tissues. Pkd2 maps to mouse Chromosome 5 and is excluded as a candidate gene for previously mapped mouse mutations resulting in a polycystic kidney phenotype. PMID- 9339381 TI - Genomic structure of a novel chloride channel gene, CLIC2, in Xq28. AB - A transcript map was previously constructed in the 1200-kb telomeric region of Xq28. One of the cDNAs, XAP121, displayed homology to a p64 bovine chloride channel and to a human chloride channel (p64CLCP, NCC27) at both the nucleotide and the peptide levels. In addition, all three sequences exhibited homologies to numerous ESTs derived from human, mouse, rat and pig. While the human NCC27 and XAP121 homologs encode small peptides of 241 and 243 amino acids, respectively, the bovine peptide has a length of 437 amino acids. This suggests that the human genes represent a novel and separate class of small chloride channels. Unlike other chloride channels, the NCC27 peptide was recently shown to localize intracellularly in the cytoplasm and nucleus. The NCC27 and XAP121 genes have thus been designated CLIC1 and CLIC2 for chloride intracellular channel genes 1 and 2, respectively. Since a direct association exists between a number of human chloride channel genes and a range of hereditary diseases, CLIC2 possibly represents a candidate for one of the many diseases linked to Xq28. To facilitate defined mutation analyses, we determined the genomic structure of the CLIC2 gene. PMID- 9339382 TI - Mapping of the gene (NRAP) encoding N-RAP in the mouse and human genomes. AB - N-RAP is a nebulin-related actin-binding protein found at the myotendon junction in skeletal muscle and at the intercalated disks in cardiac muscle. We mapped the NRAP gene to mouse chromosome 19 using interspecific crosses and to human chromosome 10 using radiation hybrid panels. Comparative analysis of the mouse and human genomes indicates that the NRAP gene is located in regions of conserved synteny between the two species. PMID- 9339383 TI - The human transaldolase gene (TALDO1) is located on chromosome 11 at p15.4-p15.5. AB - Transaldolase (TAL) is a key enzyme of the pentose phosphate pathway, which is responsible for generation of reducing equivalents to protect cellular integrity from reactive oxygen intermediates. While exons 2 and 3 are highly repetitive, the complete TAL-H gene is mapped to a single genomic locus (TALDO1(2)) by several independent approaches. Southern blot hybridization of a 827-bp 3' EcoRI fragment of the TAL-H cDNA to human-mouse somatic cell hybrid DNA localized TALDO1 to the p13-->pter region of chromosome 11. Fluorescence in situ hybridization with a 15-kb genomic fragment harboring exons 1 and 2 mapped TALDO1 to 11p15.4-p15.5. A truncated and mutated segment of TAL-H exon 5 terminating with a poly(A) tail was identified in a pseudogene locus (TALDOP1) on chromosome 1. Reverse transcriptase-PCR studies of human-mouse somatic cell hybrids revealed the presence of the functional TAL-H gene on chromosome 11 and its absence on human chromosome 1. Mapping of radiation hybrids placed TALDO1 between markers WI 1421 and D11S922 on 11p15. PMID- 9339384 TI - Assignment of the human podocalyxin-like protein (PODXL) gene to 7q32-q33. PMID- 9339385 TI - jumonji gene maps to mouse chromosome 13. PMID- 9339386 TI - Transforming growth factor-beta 2 inhibits interleukin 1 beta-induced expression of inducible nitric oxide synthase in rat renal mesangial cells. AB - OBJECTIVE AND DESIGN: We have investigated whether transforming growth factor beta 2 (TGF-beta 2) influences the expression of inducible nitric oxide synthase (iNOS). MATERIAL: Rat renal mesangial cells exposed to the inflammatory cytokine interleukin 1 beta (IL-1 beta). RESULTS: Addition of TGF-beta 2 dose-dependently suppresses IL-1 beta-induced nitrite formation. Western and Northern blot analyses of mesangial cell extracts reveal that the inhibition of IL-1 beta induced nitrite formation by TGF-beta 2 is due to decreased iNOS protein and iNOS mRNA steady state levels. Reduction of iNOS mRNA levels is due to decreased transcriptional activity of the iNOS gene under the action of TGF-beta 2 as shown by nuclear run on experiments. CONCLUSIONS: TGF-beta 2 is a potent inhibitor of iNOS expression acting by reducing the transcriptional activity of the iNOS gene. PMID- 9339387 TI - Benzydamine protection in a mouse model of endotoxemia. AB - OBJECTIVE: Previous studies have shown that benzydamine (40 mg/kg s.c.) is able to inhibit tumor necrosis factor (TNF) production and to reduce mouse lethality when administered before or concomitantly with LPS. The present study was designed to further investigate benzydamine activity against LPS-induced toxicity in terms of potency and therapeutic effects. METHODS: Female Balb/c mice were used. A dose-response curve of animal lethality versus endotoxin dose was performed (LD50 = 45 micrograms/mouse). Therapeutic effects were studied selecting the dose of LPS to achieve an LD100 (160 micrograms/mouse). Mortality was assessed daily and mice were followed for 8 days. The potential mode of action of therapeutically administered benzydamine was also investigated. TNF alpha and IL-1 beta levels were measured, at 5 h after LPS injection, both in sera and in lungs. Moreover, the drug was assayed in a TNF-dependent cytoxicity test. RESULTS: Benzydamine, administered at 20 mg/kg s.c. simultaneously with the endotoxin, significantly increased LPS LD50 up to 230 micrograms/mouse (p < 0.05). Moreover, the drug significantly protected mice against LPS-induced lethality when administered either 30 min or 4 h after endotoxin injection (p < 0.001). Benzydamine, therapeutically administered at 20 mg/kg s.c., significantly reduced TNF alpha and IL-1 beta production induced by LPS both in serum and lungs and it was shown to inhibit TNF-dependent cytoxicity on L929 cells. CONCLUSIONS: These results clearly demonstrate the therapeutic activity of benzydamine in a simple model of endotoxic shock. Available data confirm the potential role of benzydamine as an anti-cytokine agent and provide suggestions for novel therapeutic applications of this anti-inflammatory drug. PMID- 9339388 TI - Flare responses of atopic eczema patients analysed with true colour images. AB - OBJECTIVE AND DESIGN: Attenuated flare responses of atopic eczema (AE) patients to histamine are well documented, but their origin is still unknown. SUBJECTS AND METHODS: Here we studied the development of erythema after histamine iontophoresis in 12 AE patients and 12 healthy volunteers by means of a RGB camera for recording true colour images. TREATMENT: 10 mg cetirizine or placebo was administered orally 3 h before the experiment in a crossover design. RESULTS: The flare reaction was found to develop after termination of histamine iontophoresis in two phases: a first phase lasting 1-2 min in which the flare increased by about 10 mm2/s and a second phase lasting another 10-15 min characterized by a slower growth in the range of 1 mm2/s. CONCLUSIONS: Flare size was diminished in AE patients, mainly due to a slower or absent growth in the second phase. Oral application of the H1-antagonist cetirizine (Zyrtec) reduced the flare reaction in both groups of volunteers significantly, indicating that the reaction is dependent on the activation of chemosensitive nerve fibres via H1 receptors. PMID- 9339389 TI - Tumor necrosis factor measurement and use of different anticoagulants: possible interference in plasma samples and supernatants from endotoxin-stimulated monocytes. AB - OBJECTIVE AND DESIGN: Unfractionated heparin is frequently used as an anticoagulant during blood sampling and in cell culture experiments. In the present study we investigated whether heparin and other anticoagulants (citrate and EDTA) interfered with measurements of plasma tumor necrosis factor alpha (TNF alpha) concentrations or with TNF alpha release from endotoxin-stimulated monocytes. MATERIAL AND METHODS: TNF alpha was measured by a WEHI 164 bioassay in the plasma of 16 septic patients anticoagulated with heparin, citrate, or EDTA. Anticoagulants were incubated with the bioassay cell line and cell lysis was monitored. To exclude falsely low TNF alpha concentrations, anticoagulants were incubated in increasing amounts with human recombinant TNF alpha/saline solution, and rTNF alpha recovery was measured either with the WEHI 164 bioassay or an ELISA test. Further, anticoagulants were incubated with monocytes isolated from healthy volunteers and stimulated with endotoxin. Supernatants were analyzed for TNF alpha with both test systems. RESULTS: No biologically active TNF alpha was detected in the plasma with heparin anticoagulation, whereas with citrate, reproducible, TNF alpha-induced cytotoxicity was detectable in blood samples of 13 of the 16 patients. Anticoagulation with EDTA resulted in fairly high, variable and poorly reproducible TNF alpha values. Only EDTA produced falsely high values by unspecific lysis of WEHI cells. Only heparin at a concentration of 20 I.U./ml or more was found to produce falsely low values by interaction with the TNF alpha bioassay, but also with the ELISA test. In monocyte culture experiments, heparin significantly attenuated the stimulatory effect of endotoxin on TNF alpha release already at the lowest concentration tested (25 I.U./ml). CONCLUSIONS: Heparin and EDTA may have significant adverse effects on TNF alpha measurement when used for blood sampling. Citrate does not interfere with the TNF alpha bioassay or ELISA, and seems, therefore, to be the anticoagulant of choice. Due to intrinsic interactions with various cell systems (including the WEHI cell and monocytes), one should be careful in using heparin in cell culture studies in which effects of TNF alpha or of endotoxin are being studied. PMID- 9339390 TI - Glucosaminylmuramyl dipeptide (GMDP) modulates endothelial cell activities in vitro but has no effect on angiogenesis in vivo. AB - OBJECTIVE AND DESIGN: The aim of the study was to evaluate the effects of GMDP on angiogenesis in vivo and as a modulator of human umbilical vein endothelial cell proliferation, cell surface antigen expression and cell adhesion in vitro. MATERIALS: Human umbilical vein endothelial cells (HUVEC), fertilized white leghorn chicken eggs, antibodies against adhesion molecules and glucosaminylmuramyl dipeptide (GMDP). TREATMENT: GMDP [0.01-100 micrograms/ml] applied to cell cultures for 6-72 h and to the chick chorioallantoic membrane (CAM) for four days. METHODS: Angiogenic activity of GMDP in vivo was assessed using the CAM assay; HUVEC proliferation was measured by tritiated thymidine incorporation and cell cycle studies; cell surface antigen expression by indirect immunofluorescence and flow cytometry; cell adhesion by quantification of [3H] thymidine labeled leukocyte adherence to HUVEC monolayers. Statistical analysis was performed using one-way ANOVA and if necessary was followed by Duncan's multiple range test for variables. RESULTS: GMDP induced [3H]-thymidine incorporation in a concentration- and time-dependent manner (p < 0.003) and significantly increased the porportion of cells in the S phase of the cell cycle (p < 0.03). It weakly augmented the expression of ICAM-1 and CD31 but not adhesion of leukocytes to HUVEC monolayers GMDP was not angiogenic in the CAM assay. CONCLUSIONS: GMDP can modulate endothelial cell activity without the induction of angiogenesis in vivo which may have implications for its use as a therapeutic agent. PMID- 9339391 TI - Loss of cell surface microvilli on rat basophilic leukaemia cells precedes secretion and can be mimicked using the calmodulin antagonist trifluoperazine. AB - OBJECTIVE: We studied changes in cell surface morphology following treatment with secretagogue or trifluoperazine in a mast cell model. MATERIALS AND METHODS: Rat basophilic leukaemia (RBL) cells were treated with antigen and or the calmodulin antagonist, 0-50 microM trifluoperazine (TFP). The release of a secretory granule enzyme, beta-hexosaminidase, into the external medium was used as a measure of secretion. Quantitation of cell surface microvilli was determined by using a computer with input from a digitising tablet from scanning electron micrographs. Cytoskeletal proteins present in microvilli were analysed by confocal immunofluorescence. RESULTS: When RBL cells are stimulated to secrete with an antigen, the cell surface is transformed from a microvillous morphology to a ruffled one. The cell surface rearrangement preceded beta-hexosaminidase secretion: the majority of microvilli disappeared rapidly after stimulation (t1/2 of 39 s) whereas secretion can only be measured after a lag of 47 s. The calmodulin antagonist, TFP did not inhibit antigen-induced secretion or loss of microvilli, however TFP alone caused a similar loss of microvilli but was unable to stimulate or potentiate secretion. The microvilli mostly disappeared within 30 s, and a half-maximal effect occurred at approximately 8 microM TFP. Using immunofluorescence, calmodulin was localized to punctate structures on the dorsal cell surface which presumably correspond to the microvilli, and which also stained for F-actin and myosin I. CONCLUSIONS: Loss of cell surface microvilli on RBL cells precedes secretion and could reflect a cytoskeletal rearrangement which facilitates fusion of secretory granules with the membrane. It can be mimicked using trifluoperazine and we suggest it may involve calmodulin-binding components of the microvillus cytoskeleton such as myosin I. PMID- 9339392 TI - The pharmacological profile of mouse hind paw inflammation induced by staphylococcal enterotoxin type A. AB - OBJECTIVE AND DESIGN: The present paper examines the possible pharmacological mediators involved in mouse hind paw inflammation induced by staphylococcal enterotoxin type A (SEA). MATERIALS AND METHODS: The edema and the Evans blue exudation were measured in male Swiss mice (20-25 g) using methods described by Levy and Griswold, respectively. RESULTS: SEA (32 micrograms/paw) produced a biphasic, long-lasting, dose- and time-dependent edematogenic response. The acute phase edema was pronounced while the chronic edema was of a low intensity. Exudate was the principal component of the edema. The edematogenic effect of SEA appears to involve cyclooxygenase products and was dose-dependently reduced by pretreating the mice with dexamethasone, indomethacin, BW755C, WEB2086, capsaicin, diphenhydramine or cimetidine. CONCLUSIONS: These results demonstrate that SEA-induced mouse hind paw inflammation is a useful model for studying SEA mediated enterotoxemia and may be sufficiently sensitive to differentiate between the effects of SEA and those of SE type B (SEB). PMID- 9339393 TI - pH-dependent regulation of leukocyte 5-lipoxygenase activity in inflammatory exudates by albumin. AB - OBJECTIVE AND DESIGN: In order to study the regulation of cellular 5-lipoxygenase activity under inflammatory conditions, the effects of inflammatory exudates on rat leukocyte 5 lipoxygenase activity were investigated. MATERIALS: Peritoneal leukocytes and inflammatory exudates were collected from glycogen treated rats. TREATMENT: Glycogen (1 g/kg body weight, in a final volume of 3 ml PBS) was injected intraperitoneally into male Wistar rats. After 4 h, the inflammatory exudate was collected. METHODS: Rat peritoneal leukocytes were isolated and the cellular 5-lipoxygenase activity was determined by HPLC after cell stimulation with calcium ionophore A23187. RESULTS: Inflammatory exudates from glycogen treated animals strongly inhibited cellular 5-lipoxygenase activity of ionophore challenged leukocytes. Albumin was identified as the inhibitor in exudates. Inhibition of cellular 5-lipoxygenase activity by albumin was pH-dependent and was strongly increased by the alkaline pH (7.9-8.0) of the exudate. The albumin effect increased in the range of pH 7.4-8.2 where albumin undergoes a conformational change called neutral to base (N-B) transition. S-Carboxymethyl albumin had a similar activity to that of albumin, which indicated that the free SH-group at Cys-34 of albumin is not necessary for the effect. The albumin dimer showed a significantly higher inhibition than albumin and it suppressed cellular 5-lipoxygenase activity by 98%. Peptic and tryptic fragments of albumin which comprise domains I, II and II, III, respectively, were less active or inactive. Thus, an intact albumin molecule or the dimer are required for efficient inhibition of cellular 5-lipoxygenase activity. CONCLUSIONS: Our data suggest that during inflammation, albumin extravasation and changes in pH-value are involved in the regulation of the inflammatory reaction by suppression of leukotriene release. PMID- 9339394 TI - [The development of a consensus on the empiric therapy for fungal infections of patients with leukemia]. PMID- 9339396 TI - [Combination effect of fosfomycin to beta-lactam, aminoglycoside, and macrolide antibiotics against clinical isolates of Klebsiella pneumoniae]. AB - It is well known that fosfomycin (FOM) shows the combination effects with some other antibiotics. Suck effects have not been known against Klebsiella pneumoniae, however. In this report, combination effects of FOM with beta-lactam, aminoglycoside, and macrolide antibiotics were investigated against clinical isolates of both FOM-susceptible and resistant Klebsiella pneumoniae. FOM had synergistic activities with beta-lactams such as ampicillin (ABPC) and cefminox (CMNX), and macrolide antibiotics as erythromycin (EM) and midecamycin (MDM) against all strains tested. Among beta-lactams, penicillin V showed synergistic actions against FOM-susceptible strain of Tf341A and additive actions against FOM resistant 3 strains with FOM. Pheneticillin was synergistic with FOM against FOM highly susceptible strain Tf341A and the additive or nearly synergistic effects against other strains. FOM with amoxicillin was synergistic against Tf341A, and FOM-resistant strain of Tf170B and additive against other strains. While the activities of combination of FOM with kanamycin or dibekacin against FOM susceptible 2 strains were additive, those with amikacin were synergistic. Five different aminoglycosides tested showed antagonistic activities with FOM against 3 FOM-resistant strains. From these results, FOM appears to be clinically useful in treating FOM-susceptible and resistant strains of Klebsiella pneumoniae in combination of 4 antibiotics such as ABPC, CMNX, EM, and MDM. PMID- 9339395 TI - [Survey of sensitivities of clinical isolates to antibacterial agents (annual report)]. AB - Research groups were formed in 20 institutions nationwide to investigate carbapenem resistance of clinical isolates. Activities of various antibacterial agents, principally carbapenems, were tested against clinical isolates collected from these institutions. The broth microdilution method was used to determine the minimum inhibitory concentrations (MICs) of 17 antibacterial agents for 1,326 strains of 11 bacterial species isolated at the institutions between October and December 1994. The results are as follows: 1. Carbapenems exhibited strong antibacterial activities against MSSA and Streptococcus pneumoniae. Their activities against Enterococcus faecalis were comparable to that of ABPC. Carbapenems showed low activities against MRSA. 2. OFLX exhibited the greatest antibacterial activity against Haemophilus influenzae, followed by MEPM. Antibacterial activities of the other carbapenems were comparable to those of FMOX, CTM, and ABPC. 3. The carbapenems showed high activities against Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Bacteroides fragilis group. Their activities were greater than those exhibited by other beta lactam antibacterial agents. The carbapenems also exhibited stronger antibacterial activities against Serratia marcescens than the other beta-lactam antibacterial agents, but some resistant strains were detected. 4. The antibacterial activities of carbapenems against Pseudomonas aeruginosa were comparable to those of CAZ, AZT, AMK. PMID- 9339397 TI - [In vitro combination effect of vancomycin and carbapenems against carbapenem resistant MRSA]. AB - We determined in vitro combined effects of vancomycin (VCM) plus carbapenems (CRBs) on 12 methicillin-resistant Staphylococcus aureus (MRSA) which are resistant to CRBs. Combinations of VCM plus imipenem (IPM) and VCM plus panipenem (PAPM) and VCM plus meropenem (MEPM) indicated synergistic effects, fractional inhibitory concentration (FIC) indices of < or = 0.05, against 67%, 75%, 67% of the strains, respectively. Against forty two percent of strains tested, 1 MIC of VCM was equal to 1 MBC, and similarly, IPM, PAPM and MEPM had 1 MIC = 1 MBC against 42%, 67% and 75% of the strains tested, respectively. Combinations of VCM plus IPM and VCM plus PAPM and VCM plus MEPM showed synergistic effects, hence a fractional bactericidal concentration (FBC) index of < or = 0.50, against 42%, 50%, 75% of the strains, respectively, and the combination of VCM plus MEPM was most synergistic. These results suggest that combination therapy of VCM with CRB is useful for the treatment of MRSA infection in patients with renal dysfunction. PMID- 9339398 TI - [Combination use of second generation cephem and isepamicin for the treatment of post-surgical infection of the lower digestive tract]. AB - Effects of combination use of a second generation cephem and an aminoglycoside antibiotic isepamicin (ISP), for the treatment of post-surgical infections of the lower digestive tract were examined from a laboratory and clinical perspective. Thirty-three out of total 35 cases were included in the analysis of the test, while the other 2 cases did not meet the criteria for analysis. ISP was administered in combination with one of the second generation cephems among cefotiam (CTM), cefmetazole (CMZ), or cefuroxime (CXM) for 26 cases of wound infection and 7 cases of peritonitis. The overall efficacy rate was 88%; 92% in wound infection and 71% in peritonitis individually. There was no significant difference in efficacy among different groups. Bacteriological study showed the combination use of any one of the second generation cephems and ISP was very effective in all 14 cases of infections with a single species of bacterium, the efficacy rate was 100%. It was also effective in 9 out of 10 cases with mixed infections of 2 species of bacteria (90%), but effectiveness was diminished in only 6 out of 9 cases with mixed infections of 3 species (67%). Sixty-nine strains were isolated from the 35 cases, more than half of which (44 strains) were Gram-negative; 14 strains of E. coli (20%), 13 strains of E. faecalis (13%) and 6 strains of P. aeruginosa (9%). The MIC50 and MIC90 were, respectively, 1.56 micrograms/ml and 100 micrograms/ml for ISP, 6.25 micrograms/ml and 100 micrograms/ml for CTM, 12.5 micrograms/ml and 100 micrograms/ml for CMZ, and 25 micrograms/ml and 100 micrograms/ml for CXM. The MICs in any combination uses decreased synergistically according to ISP concentration. Adverse events were observed in 3 out of 35 cases, but they were not severe. The results indicated that the combination use of a second generation cephem and ISP was useful and should be one of the choices for the treatment of post-operative infections of the lower digestive tract. PMID- 9339399 TI - [Clinical efficacy in pediatrics sinusitis infections of cefditoren pivoxil granule therapy and its in vitro antibacterial activity against clinically isolated strains]. AB - We investigated the clinical efficacy in pediatrics sinusitis infections of cefditoren pivoxil granule therapy and its in vitro antibacterial activity against clinically isolated strains. The results are summarized as follows. The specimens from 343 patients were cultured and 595 strains of bacteria were isolated and identified. Oral doses of 3 and 5 mg/kg of CDTR-PI were clinically effective at high percentages, 85.1% and 89.5%, respectively, of treated patients. CDTR-PI at 3 mg/kg orally was clinically effective in 80.8% of patients with PCG intermediate S. pneumoniae (PISP) infections, 80.0% of those with PCG susceptible S. pneumoniae (PSSP) infections, 81.8% of those with H. influenzae infections and 78.3% of those with M. (B.) catarrhalis infections among the infections by major causative agents. The frequent isolates included S. pneumoniae accounting for 33.1%, H. influenzae accounting for 32.1%, M. (B.) catarrhalis accounting for 17.6% and S. pyogenes accounting 3.7% of all the isolates. PISP accounted for 16.1% of all the isolates and for 49.8% of the isolates of S. pneumoniae, and were isolated from 28.6% of the 343 patients. The isolation of PISP was frequent from children of 4 and under especially, and especially frequent from those below age 2. Of the isolates of S. pneumoniae, the biotype frequencies among PSSP were in the order of type I > type II > type III, while those among PISP were in the order of type I < type II with none of type III. Bacteriologically, an eradication rate of 89.4% was achieved with 3 mg/kg and 93.5% with 5 mg/kg of CDTR-PI. PMID- 9339400 TI - Celebrating silver: the genesis of a neuroanesthesiology society. NAS-->SNANSC- >SNACC. Neuroanesthesia Society. Society of Neurosurgical Anesthesia and Neurological Supportive Care. Society of Neurosurgical Anesthesia and Critical Care. PMID- 9339401 TI - Postoperative nausea and vomiting. A retrospective analysis in patients undergoing elective craniotomy. AB - Nausea and vomiting are important complications after craniotomy, for which there are little published epidemiologic data. We retrospectively examined the incidence of postcraniotomy nausea and vomiting to define risk factors. Medical records from 199 adults undergoing elective craniotomy were identified. Data extracted from surgery and the initial 48 hours postoperatively included gender, age, supratentorial versus infratentorial craniotomy, type of anesthesia (general versus monitored anesthesia care), intraoperative fentanyl dose, duration of anesthesia, antiemetic administration intraoperatively and postoperatively, and incidence of postoperative nausea, emesis, and opioid use. Postoperative nausea was recorded in 99 patients (50%) and emesis in 78 patients (39%). Postoperative opioids were administered to 170 patients (85%). Antiemetics were given intraoperatively to 13 patients (7%) and postoperatively to 121 patients (61%). More women (61%) than men (37%) had nausea (P = 0.001); emesis (women = 46%; men = 31%, P = 0.03); and postoperative antiemetic use (women = 69%; men = 51%, P = 0.013). The incidence of postoperative nausea (P = 0.04) and vomiting (P = 0.06) was greater in patients having infratentorial surgery. Emesis was more frequent in younger patients (P = 0.03). Postoperative nausea and vomiting were independent of anesthetic duration, fentanyl dose, or postoperative opioid use and occurred with similar frequency after general anesthesia or monitored anesthesia care. We conclude that postoperative nausea and vomiting occur frequently after craniotomy. Infratentorial surgery, female gender, and younger age are significant risk factors for this complication. PMID- 9339402 TI - Hypocapnia reverses the fentanyl-induced increase in cerebral blood flow velocity in awake humans. AB - Investigations on the effects of opioids on cerebrovascular dynamics have repeatedly demonstrated mild to moderate increases in cerebral blood flow velocity in the middle cerebral artery (CBFVMCA), cerebral blood flow, and cerebrospinal fluid pressure in humans and animals. However, the influence of hypocapnia on these fentanyl effects has not been investigated. We compared mean CBFVMCA during normo- and hypocapnia before and after administration of fentanyl (2.5 micrograms/kg i.v.) in 20 awake humans. During normocapnia (end-tidal carbon dioxide [ETCO2] 40 mmHg) fentanyl significantly increased mean CBFVMCA (60 +/- 10 cm/s vs. 81 +/- 12 cm/s [mean +/- SD]; p < 0.01), whereas during hypocapnia (ETCO2 25 mmHg) mean CBFVMCA values were identical (40 +/- 7 cm/s vs. 40 +/- 7 cm/s) before and after fentanyl administration. These results confirm previous findings that administration of fentanyl increases CBFVMCA, but, more importantly, clearly indicate that hypocapnia reverses this potentially undesirable effect. PMID- 9339404 TI - Cerebrospinal fluid passage of intravenous magnesium sulfate in neurosurgical patients. AB - Increasing evidence suggests a neuroprotective potential of magnesium sulfate (MgSO4). Only limited information about the passage of MgSO4 to the cerebrospinal fluid (CSF) is available in neurosurgical patients. However, with regard to the clinical relevance of magnesium's neuroprotective properties, quantitative data about its CSF passage are needed. The present study aims to assess the amount and the time course of magnesium's CSF passage in neurosurgical patients. To this end, 20 patients undergoing general anesthesia for neurosurgery and needing CSF drainage were included. Patients received an i.v. bolus of 60 mg/kg MgSO4. The increase in plasma and CSF magnesium concentration were measured 30, 90, and 240 min after the end of the MgSO4 infusion. These values were compared with the baseline levels taken before the start of the MgSO4 infusion. Thus, each patient served as his or her own control. Values are expressed as means +/- SD. The plasma magnesium levels were measured as follows: baseline, 0.74 +/- 0.12 mM; at 30 min, 1.24 +/- 0.1 mM (p < 0.01); at 90 min, 0.95 +/- 0.15 mM (p < 0.01), and at 240 min, 0.82 +/- 0.14 mM (p < 0.05). The CSF magnesium levels were measured as follows: baseline, 0.95 +/- 0.11 mM; at 30 min, 1.00 +/- 0.15 mM (NS); at 90 min, 1.10 +/- 0.17 mM (p < 0.01); and at 240 min, 1.13 +/- 0.19 mM (p < 0.001). We concluded that a bolus of 60 mg/kg of MgSO4 leads at least after 90 min to a significant increase in the CSF magnesium concentration. Moreover, the increase in plasma and CSF magnesium concentration is not parallel. Thus, plasma magnesium concentration cannot be used to predict the changes in CSF concentrations. PMID- 9339403 TI - High-dose fentanyl does not adversely affect outcome from forebrain ischemia in the rat. AB - Fentanyl citrate has properties, including agonism of the mu-opioid receptor and proconvulsant activity, that theoretically could pose adverse effects in ischemic brain. This study examined the effects of high-dose fentanyl on outcome in rats subjected to transient near-complete forebrain ischemia. Rats were anesthetized with halothane and surgically prepared for ischemia. In one group (fentanyl; n = 15), intravenous fentanyl (400 micrograms/kg followed by an infusion of 16 micrograms/kg/min for 20 min) was administered and halothane was discontinued. In the remaining rats (control: n = 15), halothane administration was continued and no fentanyl was given. Following 10 min of bilateral carotid artery occlusion and profound systemic hypotension, animals were maintained normocapnic, normothermic, and mildly hyperoxemic for 8 h. Four days later, histologic and neurologic outcomes were assessed. In another group of rats also administered halothane (uncontrolled recovery; n = 15), no attempt was made to control physiologic variables during recovery from ischemia. Fentanyl caused preischemic evidence of epileptoid activity but decreased the percentage of neurons that died in the CA1 sector of the hippocampus relative to control (p = 0.0005). Damage in the cortex or caudoputamen was not different from that in the control group. Rats with an uncontrolled recovery had decreased damage in the cortex (p = 0.005) and caudoputamen (p = 0.00015) relative to control. In this model of forebrain ischemia, fentanyl caused no worsening of histologic damage in the cortex or caudoputamen and decreased hippocampal CA1 injury despite major electroencephalographic activation in the immediate preischemic period. PMID- 9339406 TI - Effect of sedative and hypnotic doses of propofol on the EEG activity of patients with or without a history of seizure disorders. AB - Propofol is alleged to possess both pro- and anticonvulsant properties, leading to controversy regarding its use in patients with a history of seizures. Since propofol is administered for both sedation and hypnosis, it is important to understand the effects of low (0.5-1.0 mg/kg) and high (2-2.5 mg/kg) doses of propofol on the electroencephalogram (EEG). In this study, the hemodynamic and EEG effects of cumulative doses of propofol from 0.5 to 2.5 mg/kg i.v. were studied in 30 neurosurgical patients with or without a history of seizure disorders. While continuously recording from scalp EEG electrodes (F3, F4, C3, C4, P3, P4, O1, and O2), propofol 0.5 mg/kg was infused intravenously over 20 s. The same dose of propofol was reinjected four times at 2-min intervals, until a total dose of 2.5 mg/kg had been administered. The number and average amplitude of the EEG waves were counted and measured manually, respectively, from 80 to 90 s after beginning the injection of each dose of propofol. After lower propofol doses (0.5-1 mg/kg), the number of beta-waves increased, while alpha- and theta waves decreased significantly in all patients. However, with larger doses of propofol (total dose of 2-2.5 mg/kg), the number of beta-waves decreased and delta-waves appeared. The amplitudes of all EEG waves increased and were maintained at a higher level after administration of propofol. Spike (or sharp) waves appeared in 33% of the control patients and in 40% of the epileptic group after propofol 0.5 mg/kg and in 73% of the control and 67% of the epileptic patients after the 1.5-mg/kg dose. In the majority of patients, the spike waves disappeared when additional doses of propofol were administered. One patient in the epileptic group had an EEG-recorded and clinical grand mal seizure after propofol 1 mg/kg, but the seizure disappeared after an additional 0.5-mg/kg bolus dose was administered. The propofol-induced EEG changes appeared initially at the frontal and central EEG electrodes and subsequently at the other EEG electrodes. Overall, there were no significant differences in the spectrum of EEG changes between the two patient populations. It is concluded that propofol produces similar dose-dependent effects on EEG activity in patients with or without a history of seizure disorders. While induction of anesthesia with higher doses of propofol (> 1.5 mg/kg) in neurosurgical patients with well controlled seizure disorder is safe, smaller sedative doses should be administered with caution to epileptic patients. PMID- 9339407 TI - Combined cesarean section and clipping of a ruptured cerebral aneurysm: a case report. AB - We present a case of subarachnoid hemorrhage due to a ruptured cerebral aneurysm of the right internal carotid artery in a patient at 34 weeks of gestation (G2P1). A combined surgical procedure (cesarean section followed by clipping of the aneurysm) was performed with good maternal and fetal outcome. The differential diagnosis, the timing of neurosurgery, and the anesthetic techniques used are discussed. PMID- 9339405 TI - 7.5% hypertonic saline versus 20% mannitol during elective neurosurgical supratentorial procedures. AB - This prospective randomized clinical study was designed to compare the effects of equal volumes of 7.5% hypertonic saline solution (HS) or 20% mannitol (M) on brain bulk and lumbar cerebrospinal fluid pressure (CSFP) during elective neurosurgical procedures (aneurysm, arteriovenous malformation, or tumor). After informed consent, 50 American Society of Anesthesiologists physical Status I (ASA I) patients were randomly assigned to M (n = 25) or HS (n = 25) groups. Anesthesia protocol was identical for both, and variables monitored included mean arterial blood pressure (MAP), heart rate (HR), central venous pressure (CVP), CSF pressure (CSFP), arterial blood gases (PaCO2 30-35 mm Hg), serum sodium, potassium, and osmolality, and diuresis. The study period started before hypertonic solution administration (T0) and ended at the opening of the dura mater or 60 min after T0. Data were assessed with repeated measures analysis of variance and Student t test with Bonferroni correction (p < or = 0.05). MAP and CVP were the same in the two groups. After treatment, osmolality increased, and the increase at T15 was higher in HS-treated patients [316.6 +/- 9.3 vs. 304.0 +/ 12.0 (SD) mOsmol/kg; p < 0.001]. Sodium decreased after M and increased after HS. During the study, brain bulk was always considered satisfactory. CSFP was not different between M and HS groups and significantly decreased overtime (p = 0.0056) with no difference between treatments. The results of the present study demonstrate that hypertonic saline is as effective as mannitol in reducing the brain bulk and the CSFP during elective neurosurgical procedures under general anesthesia. PMID- 9339408 TI - Unexplained visual loss after lumbar spinal fusion. AB - Two cases of visual loss after spinal fusion surgery are described. In both cases, surgery was lengthy, the patient's head was placed in a dependent position, and hemodilution and deliberate hypotension were combined. One patient was achondroplastic, the other obese. Possible risk factors associated with ischemic optic neuropathy after anesthesia and surgery are discussed. PMID- 9339409 TI - Anesthetic implications of epilepsy, status epilepticus, and epilepsy surgery. AB - Epilepsy is a clinical paroxysmal disorder of recurring seizures, excluding alcohol or drug withdrawal seizures or such recurring exogenous events as repeated insulin-induced hypoglycemia. Epilepsy has a profound impact on each individual diagnosed with this disease. Seizures have been and are thought to arise as a result of abnormalities in (a) neural circuits, (b) excitation/inhibition balance, (c) potassium, and (d) genetic abnormalities. Therapy for epilepsy is either medical, entailing the use of a variety of antiepileptic drugs, or surgical. An urgent approach to seizure control is indicated when status epilepticus occurs. When all standard therapy fails, general anesthesia can be used to control status epilepticus. Surgery is an option in the treatment of epilepsy and requires extensive preoperative evaluation. The primary concerns for the neuroanesthesiologist anesthetizing the patient with epilepsy are the capacity of anesthetics to modulate or potentiate seizure activity and the interaction of anesthetic drugs with antiepileptic drugs. Proconvulsant and anticonvulsant properties have been reported for nearly every anesthetic. If seizure spikes are to be evoked during seizure surgery, then light anesthesia with a proconvulsant anesthetic is used. Conscious analgesia can be used for awake seizure surgery. However, if electrocorticography is not planned, then a general anticonvulsant anesthetic maintenance regimen is used. The latter technique also may be useful in patients whose anesthetic management is complicated by an incidental history of epilepsy. PMID- 9339411 TI - Direct measurement of energy metabolism in the isolated working rat heart. AB - Fatty acids and carbohydrates are the two main energy substrates used by the heart. Studies involving the regulation of these pathways in the heart have historically been hampered by a number of important technical problems. One problem is the need to provide the heart with fatty acids, which, due to their insolubility, must be delivered to the heart either bound to albumin or contained within triacylglycerol-lipoproteins. Another problem is the need to perform experiments at relevant workloads, since the work performed by the heart is a key determinant of ATP production rates. The development of the isolated working heart preparation in the 1960s has been a very powerful tool to study energy metabolism. During this golden era of cardiac energy metabolism research, a number of techniques were developed that successfully overcame these two key problems. In this article, we describe refinements to this original preparation which has allowed for simultaneous measurement of both glycolysis and glucose oxidation, or simultaneous measurements of both lactate oxidation and fatty acid oxidation. PMID- 9339410 TI - Quantitative methods for measuring the insulin-regulatable glucose transporter (Glut4). AB - This review article describes various quantitation methods for the insulin regulatable glucose transporter (Glut4). Several methods including reconstituted glucose transport, cytochalasin B binding assays, immunocytochemistry, immunoblots, ELISA, and the more recently developed exofacial labels are discussed. Since Glut4 translocates from an intracellular compartment to the plasma membrane in response to the action of insulin, it is of particular interest to measure Glut4 changes in the membrane fractions. Hence, the measurement of Glut4 commonly involves the isolation of cell membranes using subcellular fractionation in combination with one of the quantitation methods. The limitations of each quantitation method due to the use of subcellular fractionation are discussed in this article. As well, the advantages and disadvantages in terms of isoform specificity and technical difficulties of each method are presented. PMID- 9339412 TI - An isolated perfused human placental lobule model for multiple indicator dilution studies. AB - We describe a method for multiple indicator dilution studies in the isolated perfused human placental lobule developed to investigate the relationships between changes in pressure and flow and solute clearance. A peripheral lobule of a human placenta is perfused with a tissue culture-based medium and the perfusate oxygen tension, arterial and venous pressures, pH and perfusion temperature continuously monitored by a computerized system. Flow rates are readily changed. Bolus injections of vascular, extracellular and water space markers, and study compounds can be made into either maternal or fetal circulations, and precisely timed outflow fractions can be collected with computer-controlled fraction collectors, allowing simultaneous determination of concentration-time profiles of each marker. PMID- 9339413 TI - A rapid and sensitive electron-capture gas chromatographic procedure for analysis of metoprolol in rat brain and heart. AB - A procedure for analysis of metoprolol-utilizing extraction followed by derivatization with pentafluoropropionic anhydride and analysis on a gas chromatograph equipped with a fused silica capillary column, an electron-capture detector and a printer/integrator is described. Propranolol was carried through the procedure as internal standard. The pentafluoropropionyl derivative of metoprolol yields a sharp peak on the gas chromatograph, and the structure of the derivative was confirmed using combined gas chromatography-mass spectrometry. The analytical method is linear, sensitive and reproducible and has been applied to analysis of metoprolol in brain and heart from rats treated with metoprolol intraperitoneally. Pretreatment of the rats with the antidepressant desipramine prior to metoprolol administration resulted in a marked increase in levels of metoprolol in both brain and heart, indicating a pharmacokinetic drug-drug interaction between desipramine and metoprolol. PMID- 9339414 TI - Chronic renal blood flow measurement in dogs by transit-time ultrasound flowmetry. AB - To test the validity of transit-time ultrasound flowmetry for chronic measurement of renal blood flow in dogs, we compared this method with the renal clearance of para-aminohippuric acid (CPAH) (corrected for hematocrit), and with direct volumetric measurements. When flow-probes were implanted without silastic sheeting to stabilize the implant, there was significant disparity between the (within-dog) mean levels of renal blood flow estimated by flow-probe and CPAH. In contrast, when the flow-probe implants were stabilized with silicone sheeting, there was close agreement in each dog between the flow rates measured by the two methods. When flow-probes were calibrated volumetrically in situ, there was a close linear relationship between flow derived from the flow-probe and that measured volumetrically (r = 0.98 +/- 0.02). We conclude that valid, chronic measurement of renal blood flow in dogs can be achieved using transit-time ultrasound flowmetry, provided the implant is stabilized with silicone sheeting. PMID- 9339415 TI - An isolated perfused lung model with real time data collection and analysis of lung function. AB - Our primary purpose in making this report has been to describe an isolated perfused lung system which permits the real time collection and analysis of lung mechanical functioning. The distinct advantage of our system lies in its capacity for breath by breath data acquisition and analysis. In addition, because of the modular nature of the components, the system can be readily expanded or contracted depending on the type of experiment being conducted. As configured, the lung mechanic parameters of air flow, lung volume, transpulmonary pressure, pulmonary artery pressure, weight, resistance, elastance (inverse of compliance), and positive end expiratory pressure were monitored, recorded, and evaluated simultaneously throughout the experimental period. We present the results of a 3 h study with control lungs illustrating the stability of these measurements throughout the entire period. Also included is a brief discussion of 3-h studies which show a progressive loss of viability in lungs treated with the redox cycler nitrofurantoin. PMID- 9339416 TI - Urine collection in the freely moving rat: reliability for measurement of short term renal effects. AB - Studies on short-term renal responses to (pharmacological) intervention require accurate and multiple collection of urine samples. Several invasive techniques have been described for frequent urine collection of the conscious rat, each having their own limitations. No data are available about the feasibility of the spontaneously voiding, freely moving rat for this purpose. In the present study, bladder voidings of six rats were time-registered and collected separately for several days. The data show a considerable 24-h variation coefficient of both the voided volume and the bladder collection time with a poor correlation between the two parameters. Forced diuresis induced by continuous i.v. infusion (2 ml/h) increased the frequency of urine voiding and thus the time-resolution of the urine-production pattern. However, this method failed to reduce the variation coefficient of the voided volume, the collection time, and the correlation between the two parameters. The fact that variations in creatinine excretion paralleled the variation in urinary flow suggests that both phenomena are likely be due to incomplete bladder emptying. Correction for this incomplete bladder collection, using the creatinine excretion, indeed reduced the variation coefficient of sodium excretion successfully from 61 +/- 17% to 29 +/- 5% during normal diuresis and from 56 +/- 19% to 22 +/- 6% during forced diuresis. In conclusion, the spontaneously voiding, freely moving rat can be used for short term renal response studies if the collected urine samples are corrected for incomplete bladder emptying using urinary creatinine concentrations. This procedure allows the detection of changes in a urinary parameter if this exceeds a 40% deviation of the normal value. PMID- 9339417 TI - Development of an ear edema model of contact hypersensitivity to avoid false positive results due to interactions between hapten and test agents. AB - Ear edema models are regularly used for topical testing of antiinflammatory compounds. However, test compounds are usually applied simultaneously with proinflammatory agents at the same site which may result in mutual interactions. In order to avoid the occurrence of false antiinflammatory effects, a model of oxazolone-induced contact hypersensitivity has been described in which the hapten and test compound are each applied separately to only one side of the ear. By splitting and weighing the dorsal and ventral cutis of the ears, it was shown that the edemateous response of the control nonhapten side was comparable with the hapten-treated side. Some agents with antiinflammatory properties, as for example, dapsone, cimetidine, cyclosporine A, and budesonide, were tested simultaneously with oxazolone on both sides of the ear or applied separately on the dorsal and ventral ear sides, respectively. When dissolving the compounds in solutions of oxazolone, marked colorations of the test solutions were noted, indicating the occurrence of a chemical interaction. On simultaneous application at the same area, almost complete inhibition of the edemateous response was obtained for all compounds tested. In contrast, when applied separately, only budesonide appeared to exhibit antiinflammatory activity. The results indicate that the proposed model can be used to avoid the occurrence of interactions between oxazolone, and possibly other sensitizers, and substances that are being evaluated for topical antiinflammatory activity. By use of this model spurious antiinflammatory activity can be detected. PMID- 9339418 TI - Which indexes of filling behavior should be used to characterize left ventricular diastolic function when changes in heart rate and atrioventricular delay occur? AB - The routine use of the peak early-to-peak atrial velocity, early velocity integral-to-atrial velocity integral, and early velocity integral-to-the total filling velocity integral ratios are limited because they are influenced by heart rate and atrioventricular delay. Hence, we sought to establish whether these ratios could be normalized to account for the differences in cycle length (RR interval) and diastolic filling period when heart rate and atrioventricular delay were altered in 18 patients with programmable dual-chamber pacemakers. We further explored whether these and other parameters of the mitral velocity profile could be used to characterize the mitral filling pattern during isoproterenol and methoxamine infusions-interventions that are likely to change both heart rate and left ventricular filling. The early velocity integral-to-atrial velocity integral and early velocity integral-to-the total filling velocity integral ratios were more sensitive to minor variations in heart rate and atrioventricular delay than the peak early-to-peak atrial velocity ratio. The early velocity integral-to atrial velocity integral and early velocity integral-to-total filling velocity integral ratios could not be normalized to account for differences in RR interval or diastolic filling period, whereas the peak early-to-peak atrial velocity ratio became less sensitive to changes in heart rate when it was divided by the RR interval, or diastolic filling period, or square root of diastolic filling period. Because the diastolic filling period is affected by atrioventricular delay independent of changes in the RR interval, these ratios were also corrected for the functional cycle length, defined as the interval from R-wave of the electrocardiogram to the end of the diastolic filling period. When corrected for either the functional cycle length or diastolic filling period or square root of diastolic filling period, only the peak early-to-peak atrial velocity ratio became less sensitive to variations in the atrioventricular delay. The ratio of diastolic filling period expressed as a proportion of RR interval or functional cycle length changed significantly when heart rate and atrioventricular delay were altered and did not improve when diastolic filling period was divided by the square root of RR or square root of functional cycle length. However, when the square root of diastolic filling period was divided by the RR interval or functional cycle length, the effects of heart rate and atrioventricular delay were not apparent. Of all the ratios, the ratio of square root of diastolic filling period expressed as a proportion of RR interval or functional cycle length was the most useful to differentiate the confounding effects of heart rate (+/-atrioventricular delay) from the effects of isoproterenol and methoxamine on left ventricular filling. Hence, this ratio appeared to be a heart rate- and atrioventricular delay-independent index of left ventricular diastolic function. PMID- 9339419 TI - Right and left isovolumic ventricular relaxation time intervals compared in patients by means of a single-pulsed Doppler method. AB - Right and left isovolumic ventricular relaxation time intervals measurements were obtained as follows: from the peak R wave on the electrocardiogram to either the mitral or the tricuspid pulsed Doppler flow trace onset minus the R to end ejection zero flow crossing of the subaortic (left side) or pulmonary (right side) D flow trace time interval. A ratio was calculated as a percent difference duration between both isovolumic ventricular relaxation time intervals. The aim was to compare isovolumic ventricular relaxation time interval values in 42 healthy controls and to study the changes induced by heart diseases in 27 patients with (1) controlled hypertension without left ventricular hypertrophy, (2) hypertrophic cardiomyopathy, and (3) Cor pulmonale. Mean values of isovolumic ventricular relaxation time intervals significantly differed at paired and unpaired studies, with right isovolumic ventricular relaxation time intervals shorter than those of the left side in all groups (p < 0.001) except for patients with Cor pulmonale. Isovolumic ventricular relaxation time intervals did not correlate with heart rate and moderately correlated with left ventricular mass and age. No significant difference was found between healthy controls and patients with controlled hypertension. Significant changes were found in patients with hypertrophic cardiomyopathy and Cor pulmonale versus healthy controls for both isovolumic ventricular relaxation time intervals. However, significant changes in the ratio were only found in patients with Cor pulmonale (p < 0.005) because of abnormal similar values for both isovolumic ventricular relaxation time intervals. This Doppler method enabled, for the first time, serial comparison of isovolumic ventricular relaxation time intervals with a homologous method. Both isovolumic ventricular relaxation time intervals significantly lengthened with age and with left ventricular indexed mass, but their ratio remained insignificantly changed except for patients with Cor pulmonale. The concomitant right and left isovolumic ventricular relaxation time intervals lengthening in patients with hypertrophic cardiomyopathy and Cor pulmonale suggests interdependence of both ventricles through the septum. This makes recommendable systematic comparison of both sides. The calculation of a ratio, free from the effect of factors intervening on isovolumic ventricular relaxation time intervals, may, in addition, be of diagnostic help. PMID- 9339420 TI - Mid-systolic drop in left ventricular ejection velocity in obstructive hypertrophic cardiomyopathy--the lobster claw abnormality. AB - In many patients with obstructive hypertrophic cardiomyopathy, an abrupt mid systolic drop in left ventricular ejection velocity can be detected. We analyzed 27 patients with obstructive hypertrophic cardiomyopathy who had 43 echocardiographic examinations (mean gradient 53 +/- 6 mm Hg). Exams showing a mid-systolic drop had higher mean outflow tract pressure gradients (90 +/- 6 compared with 29 +/- 4 mm Hg, p < 0.001). After medical elimination of obstruction, the mid-systolic drop was no longer seen. We measured 105 pulsed wave Doppler tracings in the left ventricular cavity and compared them with 90 continuous-wave tracings through the outflow tract. There was a close temporal correlation between the nadir of the left ventricular velocity drop and the peak continuous-wave left ventricular outflow tract velocity (r = 0.99). There was also a close temporal correlation between the onset of the fall in pulsed velocity and the onset of M-mode mitral-septal contact (r = 0.95). CONCLUSIONS: The mid-systolic drop in left ventricular velocity is due to impedance to ejection and provides evidence of true obstruction. As left ventricular ejection velocity falls to its mid-systolic nadir because of impedance of ejection, velocity downstream in the left ventricular outflow tract actually rises to its peak. This disparity in the two velocities, deceleration in the left ventricular cavity and acceleration in the left ventricular outflow tract, indicates that the outflow orifice is progressively narrowed over time as the mitral valve is forced into the septum by the rising pressure difference. The obstruction phase is best described as a time-dependent, amplifying feedback loop. The orifice narrows over time because of the rising pressure difference; the pressure difference rises over time because of the narrowing orifice. PMID- 9339421 TI - Morphology and dynamic change of discrete subaortic stenosis can be imaged and quantified with three-dimensional transesophageal echocardiography. AB - This report describes three-dimensional transesophageal echocardiographic findings in three consecutive patients with discrete subaortic stenosis. The discrete subaortic stenosis lesions included a circumferential, a remnant crescent, and a broken fibrotic subaortic membrane. The lesions were best imaged by using a three-dimensional transesophageal echocardiography-generated "aortotomy view" of the left ventricular outflow tract immediately below the plane of the aortic valve. The three-dimensional images correlated well with surgical and pathologic findings. The three-dimensional surface areas of the left ventricular outflow tract at the level of discrete subaortic stenosis during systole (0.8 +/- 0.5 cm2) and diastole (1.7 +/- 0.7 cm2) were measured by planimetry of the three-dimensional transesophageal echocardiographic images. The novel "aortotomy view" offered by three-dimensional transesophageal echocardiography provided direct visualization and quantification of discrete subaortic stenosis in a dynamic fashion. In summary, three-dimensional transesophageal echocardiography can accurately display and quantify discrete subaortic stenosis and could be a new clinically useful tool for assessing discrete subaortic stenosis and guiding surgical and transcatheter interventions. PMID- 9339422 TI - Acquired dynamic left ventricular outflow tract obstruction complicating acute anterior myocardial infarction: serial echocardiographic and clinical evaluation. AB - We describe three cases of dynamic outflow obstruction complicating acute anterior myocardial infarction. Serial echocardiography suggests the intraventricular gradient results from basal hyperkinesis, the latter being a reciprocal response to the apical wall motion abnormality. PMID- 9339423 TI - Screening for abdominal aortic aneurysm during transthoracic echocardiography in a hypertensive patient population. AB - This study was undertaken to determine the utility of transthoracic echocardiography as a screening test for occult abdominal aortic aneurysm in hypertensive patients older than 50 years of age. Longitudinal and transverse images of the abdominal aorta were obtained during the subcostal portion of the transthoracic echocardiogram. Abdominal aortic aneurysm was defined as an abdominal aortic dimension (antero-posterior or lateral) > or = 3.0 cm. Exclusion criteria included prior abdominal aortic aneurysm repair, known abdominal aortic aneurysm, or inadequate images of the abdominal aorta (nine patients). Two hundred patients (107 men, 93 women; mean age 71 years, range 51 to 92 years) met the study inclusion criteria. An occult abdominal aortic aneurysm was identified in 13 patients (6.5%). Sixty-nine percent of the abdominal aortic aneurysm patients were men, with a mean age of 73 years and a mean duration of hypertension of 11 years. Seventy-seven percent had a history of tobacco use, and 15% had a positive family history of abdominal aortic aneurysm. All aneurysms were infrarenal in location, with abdominal aortic aneurysm diameter ranging from 3.0 to 5.2 cm (mean 3.9 cm). Laminated thrombus was present in six patients (46%), and in one patient a right common iliac artery aneurysm was also detected. Imaging of the abdominal aorta during transthoracic echocardiography required an average of 6.7 minutes (range 4 to 10 minutes). In conclusion, the abdominal aorta could be accurately imaged in the majority of patients (96%) undergoing transthoracic echocardiography in this study. The incidence of occult abdominal aortic aneurysm in hypertensive patients older than 50 years of age is significant (6.5%). Screening for occult abdominal aortic aneurysm in this patient population should be a routine extension of the transthoracic echocardiogram. PMID- 9339424 TI - Transnasal transesophageal echocardiography. AB - Transesophageal echocardiography has been used as a diagnostic tool in the critical care unit. However, long-term serial evaluation of ventricular function with transesophageal echocardiography is difficult because of the current probe sizes and intolerance to prolonged oral intubation. We performed 139 intubations (64 oral and 75 transnasal) with a new prototype probe in 128 patients referred for transesophageal echocardiography. Transnasal intubation with the prototype probe was possible in 63/75 attempts. Oral intubation was successful in all 64 attempts. Patients tolerated transnasal intubation well when mildly sedated or awake. Two-dimensional echocardiographic views obtained with the nasal probe were similar to those obtained with a standard monoplane probe. Image quality was rated as good or acceptable in nearly all cases. Transgastric short-axis imaging of the left ventricle combined with acoustic quantification provided stable left ventricular area waveforms. Using custom developed software we showed the feasibility of monitoring left ventricular performance with minimal probe adjustment while graphically displaying and updating left ventricular area and fractional area change. Thus, transesophageal echocardiography with a prototype miniaturized monoplane probe passed transnasally is feasible, safe, and well tolerated by patients. This probe provides excellent two-dimensional echocardiographic images and may allow long-term echocardiographic monitoring of ventricular performance. PMID- 9339425 TI - Transesophageal echocardiography: unusual case of anomalous pulmonary venous connection to the azygos vein. AB - Partial anomalous pulmonary venous connection, a rare congenital anomaly, most commonly involves the right lung, with one or more pulmonary veins anomalously connecting most frequently to the superior vena cava and less commonly to the right atrium or inferior vena cava. This article describes an unusual case of anomalous pulmonary venous connection of the right lung to the azygos vein in an adult. This anomaly was clearly delineated with angiography, computed tomography of the chest, and transesophageal echocardiography. The transesophageal echocardiographic features of the anomaly are described as a means to prevent further diagnostic misinterpretation. PMID- 9339426 TI - Saccular outpouchings of an ascending aortic aneurysm: transesophageal echocardiographic appearance. AB - The differential diagnosis of cavities in the ascending aorta includes pseudoaneurysms, intimal flaps, and abscesses. We describe the transesophageal echocardiographic and pathologic appearance of a fusiform ascending aortic aneurysm that contained atypical outpouchings that were initially confused with an intimal flap. Awareness of this unreported abnormality and its echocardiographic features will avoid the misdiagnosis of more serious aortic pathology such as acute aortic dissection or infective endocarditis. PMID- 9339427 TI - Transesophageal echocardiographic identification of a retrograde dissection of the ascending aorta caused by inadvertent cannulation of the common carotid artery. AB - Retrograde aortic dissections can be a complication of vascular procedures. We describe a case of an inadvertent cannulation of the right common carotid artery during an attempt at inserting a pulmonary artery catheter. This resulted in dissection of the right common carotid, subclavian, and innominate arteries. Transesophageal echocardiography was able to visualize a retrograde dissection extending back into the ascending aorta. PMID- 9339429 TI - Ruptured sinus of Valsalva aneurysm simulating endocarditis. AB - A previously healthy, 31-year-old man was evaluated in the emergency department after being violently assaulted. A harsh, continuous murmur was noted on physical examination. Transthoracic and transesophageal echocardiograms were interpreted as showing a ruptured sinus of Valsalva aneurysm with a shunt into the right atrium and a tricuspid valve vegetation. The patient was treated with antibiotics for presumed endocarditis. Subsequent echocardiographic and surgical evaluation showed no evidence of past or present endocarditis. Rather, the sinus of Valsalva aneurysm and rupture gave the appearance of a valvular mass. This report shows some of the potential pitfalls in the delineation of abnormalities related to sinus of Valsalva aneurysms and rupture. PMID- 9339428 TI - Transesophageal echocardiography-guided transvenous biopsy of a cardiac sarcoma. AB - Transvenous endomyocardial biopsy is a well established procedure to diagnose rejection after heart transplantation as well as to assess for other cardiomyopathic processes. However, it is rarely used to obtain samples of unidentified cardiac masses. We report a case of a primary cardiac sarcoma in which the histologic diagnosis was provided by transesophageal echocardiography guided transvenous biopsy. This procedure is accurate and can avoid the potential risk of a diagnostic thoracotomy. PMID- 9339430 TI - Pulmonary embolism from in situ right atrial thrombus after coronary artery bypass surgery. AB - Pulmonary embolism after cardiac surgery is attributed to embolization from thrombus within the deep venous system. We report two cases of pulmonary embolism after coronary artery bypass surgery in which transesophageal echocardiography detected in situ right atrial thrombus. The right atrium should be screened for thrombus in patients who have pulmonary embolism after cardiac surgery. PMID- 9339431 TI - Late pulmonary venous complications after lung transplantation. AB - Although abnormalities of the pulmonary venous anastomosis in the early postoperative period after lung transplantation have been reported from several centers, late complications related to the pulmonary venous anastomosis have not been described. In the present study, we describe the clinical and transesophageal echocardiographic findings of pulmonary vein anastomotic complications in two patients at 1.9 and 2.3 years after lung transplantation. Both pulmonary venous abnormalities, stenosis in the first instance and thrombosis in the second instance, impaired venous outflow on the affected side causing unilateral edema and pleural effusion. Pulmonary venous abnormalities late after lung transplantation can mimic allograft rejection, opportunistic infection, or heart failure and are best diagnosed by transesophageal echocardiography. PMID- 9339432 TI - Myocardial infarction as a complication of dobutamine stress echocardiography. AB - Only few cases of myocardial infarction complicating dobutamine stress echocardiography have been reported. We observed a 42-year-old woman in whom acute inferior wall infarction developed 10 minutes after discontinuation of dobutamine stress echocardiography with up to 20 micrograms/kg/min dobutamine. The right coronary artery, which had had a 50% stenosis in the prior angiography, showed proximal complete occlusion on the immediately performed recatheterization. Thrombolysis in myocardial infarction study flow grade 3 was rapidly accomplished by intracoronary thrombolysis with recombinant tissue type plasminogen activator. For recurrent unstable angina, the patient received coronary bypass grafting on the same day. The case shows that myocardial infarction not preceded by regional wall motion abnormalities is a possible complication of dobutamine stress echocardiography. Post-test monitoring even after negative tests is recommended. PMID- 9339433 TI - Echocardiography for the assessment of myocardial viability. AB - The identification of viable myocardium in the setting of acute myocardial infarction or chronic coronary artery disease with reduced left ventricular function has important prognostic and therapeutic implications. Many noninvasive methods have been used to assess viability, and recently, dobutamine stress echocardiography has been studied for this purpose. Dobutamine stress echocardiography is a safe, accessible, and relatively inexpensive technique. Moreover, its accuracy for detecting viability approaches that of positron emission tomography and thallium scintigraphy. In addition to dobutamine stress echocardiography, other echocardiographic techniques, such as myocardial contrast echocardiography and dipyridamole stress echocardiography, are being developed to delineate viability. In the future, echocardiographic methods may identify viability with enough accuracy to allow us to better select patients for revascularization procedures when the indications are otherwise unclear. PMID- 9339434 TI - Novel index relating both isovolumetric contraction time and isovolumetric relaxation time to ejection time. PMID- 9339435 TI - Three-dimensional flow-independent peripheral angiography. AB - A magnetization-prepared sequence, T2-Prep-IR, exploits T1, T2, and chemical shift differences to suppress background tissues relative to arterial blood. The resulting flow-independent angiograms depict vessels with any orientation and flow velocity. No extrinsic contrast agent is required. Muscle is the dominant source of background signal in normal volunteers. However, long-T2 deep venous blood and nonvascular fluids such as edema also contribute background signal in some patients. Three sets of imaging parameters are described to address patient specific contrast requirements. A rapid, spiral-based, three-dimensional readout is utilized to generate high-resolution angiograms of the lower extremities. Comparisons with x-ray angiography and two-dimensional time-of-flight angiography indicate that this flow-independent technique has unique capabilities to accurately depict stenoses and to visualize slow flow and in-plane vessels. PMID- 9339436 TI - Real-time interactive MRI on a conventional scanner. AB - A real-time interactive MRI system capable of localizing coronary arteries and imaging arrhythmic hearts in real-time is described. Non-2DFT acquisition strategies such as spiral-interleaf, spiral-ring, and circular echo-planar imaging provide short scan times on a conventional scanner. Real-time gridding reconstruction at 8-20 images/s is achieved by distributing the reconstruction on general-purpose UNIX workstations. An X-windows application provides interactive control. A six-interleaf spiral sequence is used for cardiac imaging and can acquire six images/s. A sliding window reconstruction achieves display rates of 16-20 images/s. This allows cardiac images to be acquired in real-time, with minimal motion and flow artifacts, and without breath holding or cardiac gating. Abdominal images are acquired at over 2.5 images/s with spiral-ring or circular echo-planar sequences. Reconstruction rates are 8-10 images/s. Rapid localization in the abdomen is demonstrated with the spiral-ring acquisition, whereas peristaltic motion in the small bowel is well visualized using the circular echo planar sequence. PMID- 9339437 TI - Gradient moment smoothing: a new flow compensation technique for multi-shot echo planar imaging. AB - This work identifies an additional source of phase error across ky in multi-shot echo-planar imaging resulting from flow or motion along the phase-encoding direction. A velocity-independent flow compensation technique, gradient moment smoothing, is presented that corrects this error by forcing the phase to have smooth quadratic behavior. The correction is implemented, without compromising scan time, by changing the first moment of a bipolar prephaser pulse on a shot-by shot basis. In phantom and in vivo experiments, gradient moment smoothing effectively eliminates ghosting and signal loss due to phase-encoding flow. When used in conjunction with a "flyback" echo-planar readout, which compensates for flow in the frequency-encoding direction, gradient moment smoothing renders multi shot echo-planar imaging relatively insensitive to in-plane flow. This can make multi-shot echo-planar imaging a viable technique for accurately imaging in-plane flow and may desensitize it to the otherwise serious problem of in-plane motion. PMID- 9339438 TI - Manganese ion enhances T1-weighted MRI during brain activation: an approach to direct imaging of brain function. AB - Present techniques for functional MRI rely on detecting changes in hemodynamics that result as a consequence of brain activation. It would be useful if MRI techniques could be developed that enable imaging of a parameter directly related to neuronal activity. Influx of calcium into neurons is necessary for release of neurotransmitters. Divalent manganese ions (Mn2+) can enter cells through voltage gated calcium channels and Mn2+ is paramagnetic. Mn2+ accumulation in brain due to activation should alter relaxation times offering an approach to sensitize MRI to calcium influx in the brain. To test this idea, T1-weighted MRI was obtained from the rat brain in the presence of a continuous intravenous infusion of 3.6 mumol/min MnCl2. In the anesthetized rat brain, signal enhancement was detected in regions corresponding to ventricles. Activation of the brain with glutamate led to increase in MRI signal intensity in the brain to 238 +/- 23% of the original. This increase in signal was dependent on the presence of MnCl2 and was not due to changes in blood flow. It was necessary to break the blood brain barrier with mannitol to make Mn2+ accessible to the active sites for efficient detection. Enhancement of MRI signal in the brain was also detected with decreasing anesthesia and with somatosensory stimulation. Due to the slow clearance of Mn2+ from the stimulated region of the brain, MRI enhancement could also be detected after stimulation that occurred on awake, behaving rats outside the magnet. These data indicate that MnCl2 shows potential as a MRI contrast agent that is directly sensitive to brain activation. PMID- 9339439 TI - Detection of dopaminergic neurotransmitter activity using pharmacologic MRI: correlation with PET, microdialysis, and behavioral data. AB - The metabolic activation resulting from direct dopaminergic stimulation can be detected using auto-radiography, positron emission tomography (PET) or, potentially, fMRI techniques. To establish the validity of the latter possibility, we have performed a number of experiments. We measured the regional selectivity of two different dopaminergic ligands: the dopamine release compound D-amphetamine and the dopamine transporter antagonist 2 beta-carbomethoxy-3 beta (4-fluoropheny) tropane (CFT). Both compounds led to increased signal intensity in gradient echo images in regions of the brain with high dopamine receptor density (frontal cortex, striatum, cingulate cortex > > parietal cortex). Lesioning the animals with unilaterally administered 6-hydroxydopamine (6-OHDA) led to ablation of the phMRI response on the ipsilateral side; control measurements of rCBV and rCBF using bolus injections of Gd-DTPA showed that the baseline rCBV and rCBF values were intact on the lesioned side. The time course of the BOLD signal changes paralleled the changes observed by microdialysis measurements of dopamine release in the striatum for both amphetamine and CFT; peaking at 20-40 min after injection and returning to baseline at about 70-90 min. Signal changes were not correlated with either heart rate, blood pressure or pCO2. Measurement of PET binding in the same animals showed an excellent correlation with the phMRI data when compared by either measurements of the number of pixels activated or percent signal change in a given region. The time course for the behavioral measurements of rotation in the 6-OHDA lesioned animals correlated with the phMRI. These experiments demonstrate that phMRI will become a valuable, noninvasive tool for investigation of neurotransmitter activity in vivo. PMID- 9339440 TI - Gradient, high-resolution, magic-angle spinning nuclear magnetic resonance spectroscopy of human adipocyte tissue. AB - The recently developed technique of gradient, high-resolution magic-angle spinning NMR (g-hr-MAS-NMR) spectroscopy was applied to the study of ex vivo human lipoma and liposarcoma tissue. Compared with conventional 1H-NMR, the g-hr MAS method yielded a large improvement in spectral resolution and permitted the detection of metabolite resonance's in a well-differentiated liposarcoma that was not observed in spectra of similar samples obtained using nonspinning NMR methods. These findings suggest that g-hr-MAS-NMR spectroscopy provides a key improvement in spectral quality for ex vivo lipoma and liposarcoma tissue thereby permitting a more precise determination of tissue metabolite composition than conventional nonspinning NMR methods. PMID- 9339441 TI - STAR-HASTE: perfusion imaging without magnetic susceptibility artifact. AB - A novel magnetic resonance imaging technique (STAR-HASTE) based on pulsed arterial spin labeling using a single shot acquisition method is described for perfusion imaging. The method is similar to EPISTAR in using STAR (Signal Targeting with Alternating Radiofrequency) technique for pulsed radiofrequency labeling of inflowing blood, but uses a half-Fourier single shot turbo spin-echo (HASTE) sequence for data acquisition instead of echo-planar imaging (EPI). Our preliminary studies show that STAR-HASTE permits perfusion imaging to be performed without many of the artifacts encountered with other imaging methods based on EPI acquisition. The novel method not only provides similar perfusion information to that obtained by EPISTAR, as demonstrated in the functional brain imaging study, but also eliminates magnetic susceptibility artifacts and image distortion commonly observed in EPI images. Furthermore, this technique can be readily implemented in MR systems without EPI capability. PMID- 9339442 TI - In vivo imaging of a stable paramagnetic probe by pulsed-radiofrequency electron paramagnetic resonance spectroscopy. AB - Imaging of free radicals by electron paramagnetic resonance (EPR) spectroscopy using time domain acquisition as in nuclear magnetic resonance (NMR) has not been attempted because of the short spin-spin relaxation times, typically under 1 microsecond, of most biologically relevant paramagnetic species. Recent advances in radiofrequency (RF) electronics have enabled the generation of pulses of the order of 10-50 ns. Such short pulses provide adequate spectral coverage for EPR studies at 300 MHz resonant frequency. Acquisition of free induction decays (FID) of paramagnetic species possessing inhomogenously broadened narrow lines after pulsed excitation is feasible with an appropriate digitizer/averager. This report describes the use of time-domain RF EPR spectrometry and imaging for in vivo applications. FID responses were collected from a water-soluble, narrow line width spin probe within phantom samples in solution and also when infused intravenously in an anesthetized mouse. Using static magnetic field gradients and back-projection methods of image reconstruction, two-dimensional images of the spin-probe distribution were obtained in phantom samples as well as in a mouse. The resolution in the images was better than 0.7 mm and devoid of motional artifacts in the in vivo study. Results from this study suggest a potential use for pulsed RF EPR imaging (EPRI) for three-dimensional spatial and spectral spatial imaging applications. In particular, pulsed EPRI may find use in vivo studies to minimize motional artifacts from cardiac and lung motion that cause significant problems in frequency-domain spectral acquisition, such as in continuous wave (cw) EPR techniques. PMID- 9339443 TI - Calculations and experimental studies of the lineshape of the lactate doublet in PRESS-localized 1H MRS. AB - Accurate quantification of NMR metabolites by spectral modeling requires assumed lineshape functions. For singlet resonances, a combination of Lorentzian and Gaussian lineshapes is sufficient, but for weekly J-coupled resonances such as lactate, more complex lineshapes are necessary. In this work, the lactate lineshape is calculated for the PRESS sequence using standard RF pulses, and compared with experimental measurements. A similar comparison is made for PRESS localized spectroscopic imaging, in which the lineshape varies from voxel to voxel across the field of view. These observations have important implications for the quantification of lactate in experimental and human studies. PMID- 9339444 TI - Quantitative measurements of regional cerebral blood volume using MRI in rats: effects of arterial carbon dioxide tension and mannitol. AB - A three-dimensional (3D) T1-weighted sequence was used to acquire high spatial resolution whole brain images in rats before and after the injection of an intravascular contrast agent. These T1-weighted images were used to estimate regional cerebral blood volume (rCBV) as a percentage of blood volume in each voxel. Ventilation was manipulated to investigate the effects of altered arterial carbon dioxide tension (PaCO2) on rCBV. In addition, different doses of a hypertonic mannitol solution were used to investigate the sensitivity of the proposed method in a serial monitoring paradigm. An rCBV of 2.40% +/- 0.34% was obtained before any physiological manipulation, in good agreement with literature values using alternative techniques. Using this method, it was found that there exists a linear relationship between PaCO2 and rCBV (R2 = 0.77) and that rCBV increased in a dose and time dependent fashion in mannitol-treated rats. High signal-to-noise was available due to the substantial increase in blood signal from the intravascular contrast agent. PMID- 9339445 TI - Quantification and reduction of ghosting artifacts in interleaved echo-planar imaging. AB - A mathematical analysis of ghosting artifacts often seen in interleaved echo planar images (EPI) is presented. These artifacts result from phase and amplitude discontinuities between lines of k-space in the phase-encoding direction, and timing misregistrations from system filter delays. Phase offsets and time delays are often measured using "reference" scans, to reduce ghosting through postprocessing. From the expressions describing ghosting artifacts, criteria were established for reducing ghosting to acceptable levels. Subsequently, the signal to-noise ratio (SNR) requirements for estimation of time delays and phase offsets, determined from reference scans, was evaluated to establish the effect of estimation error on artifact reduction for interleaved EPI. Artifacts resulting from these effects can be reduced to very low levels when appropriate reference scan estimation is used. This has important implications for functional MRI (fMRI) and applications involving small changes in signal intensity. PMID- 9339446 TI - In vivo 1H-NMR microimaging with respiratory triggering for monitoring adoptive immunotherapy of metastatic mouse lymphoma. AB - The metastatic ESb-MP murine lymphoma in DBA/2 mice has been used as a model for investigating metastatic disease and its cure by adoptive immunotherapy (ADI) as monitored by in vivo multislice spin-echo 1H NMR microimaging at 7 T. isoflurane inhalation anesthesia facilitated long measurement sessions, and respiratory gating with a fiber-optic sensor greatly reduced motional artifacts. With T2 weighting (TR = 2 s, TE = 30 ms) mean signal-to-noise ratios of 30 and 15 for kidney and liver, respectively, were achieved in 20 min (100-micron pixels, 1-mm slices, 25-mm field of view). Without the use of contrast agents, metastases with diameters > or = 0.3 mm in the imaged plane could be detected as hyperintense lesions in kidney (contrast ratio ca. 1.4) and liver (contrast ratio ca. 2) with a confidence level of > 98%. For the first time the complete eradication of late stage macroscopic metastases by ADI could be demonstrated noninvasively by MRI. PMID- 9339448 TI - Contour-based registration technique to differentiate between task-activated and head motion-induced signal variations in fMRI. AB - Data acquired using functional magnetic resonance imaging are often contaminated by head motion. As a result, optimal information regarding task-induced (or resting-state) signal changes cannot be extracted. Intensity-based registration methods, including intensity correlation or minimum intensity variance techniques, are widely used to register two or more images. It is shown here that intensity-based registration cannot accurately register two or more images in the presence of local intensity changes arising from functional magnetic resonance, fMRI, signals. In this paper, we present a contour-based technique that can be used not only for a more robust registration, but also to help differentiate between task-induced and motion-induced signal changes. Results obtained using both phantom and human brain images demonstrate advantages of this technique compared with a conventional intensity registration technique. PMID- 9339447 TI - Noise suppression digital filter for functional magnetic resonance imaging based on image reference data. AB - The central decision in every functional magnetic resonance imaging (fMRI) experiment is whether pixels in brain tissues are showing activation in response to neural stimulus or as a result of noise. Images are degraded not only by random (e.g., thermal) noise, but also by structured noise due to MR system characteristics, cardiac and respiratory pulsations, and patient motion. A novel digital filter has been developed to suppress cardiac and respiratory structured noise in fMRI images, using estimates of structured and random noise power spectra obtained directly from the images. It is an adaptive filter based on stationary noise statistics, and is equivalent in form to a Wiener filter. A mathematical model of the filtering process was developed to understand how the strength and distribution of structured and random noise power influenced filter performance. The filter was tested using images from an auditory activation study in ten subjects. In subjects whose structured noise power was localized to a relatively narrow frequency range, a strong relationship was found, both experimentally (R = 0.975, P < 0.0004 for H0: R = 0) and using the model, between filter performance and the level of structured noise power contaminating the experiment frequency. The filter significantly reduced the rate of false-positive activations in the subset of subjects whose experiment frequency was relatively heavily contaminated by structured noise. Notch filters, that simply eliminate unwanted frequencies, performed poorly in all subjects. Unlike the proposed Wiener filter, these filters did not suppress structured noise power at the experiment frequency that contributes to false-positive activations. PMID- 9339449 TI - Dynamic shimming for multi-slice magnetic resonance imaging. AB - Dynamic shimming has been implemented in three pulse sequences on a commercial GE Signa 1.5-T imaging system. Multi-slice field maps are acquired before the imaging sequence, and linear shim terms and center frequencies are calculated for each slice. During the imaging scan, the linear shim terms and center frequency are set before each pulse sequence repetition according to the current slice. Acquisition of multi-slice field maps and calculation of shim terms and center frequency for each slice are accomplished in a matter of seconds. Pulse sequences require only minimal modification to add dynamic shimming capability. Results are shown for a fat saturation spin-echo sequence, a single-shot echo-planar gradient recalled echo sequence, and a spiral acquisition gradient-recalled echo sequence. In all cases, dynamic shimming with shim currents and center frequency optimized for each slice is shown to give better results than constant shim currents and a single center frequency optimized for the entire volume of interest. PMID- 9339450 TI - Diffusion-weighted imaging of the human optic nerve: a new approach to evaluate optic neuritis in multiple sclerosis. AB - The apparent diffusion coefficient (ADC) in the optic nerve was measured from diffusion-weighted magnetic resonance imaging using an intravoxel incoherent motion (IVIM) sequence. The subjects were seven normal volunteers and eight patients with multiple sclerosis (MS) with a total of four optic nerves with acute neuritis and nine nerves with chronic neuritis. The mean ADC (4.18 +/- 1.13 x 10(-3) mm2/s, n = 9) in the optic nerves with chronic neuritis was significantly higher than that in normal volunteers (1.56 +/- 0.675 x 10(-3) mm2/s, n = 14) and that in the nerves with acute neuritis (0.94 +/- 0.43 x 10(-3) mm2/s n = 4) (P < 0.001). The ADC is useful in assessing MS foci in the optic nerves. PMID- 9339451 TI - A method to measure arbitrary k-space trajectories for rapid MR imaging. AB - A method to measure arbitrary k-space trajectories was developed to compensate for nonideal gradient performance during rapid magnetic resonance (MR) imaging with actively or nonactively shielded gradients at a magnetic field strength of 4.1 T. Accurate MR image reconstruction requires knowledge of the k-trajectory produced by the gradient waveforms during k-space sampling. Even with shielded gradients, residual eddy currents and imperfections in gradient amplifier performance can cause the true k-space trajectory to deviate from the ideal trajectory. The k-space determination was used for spiral gradient-echo imaging fo the human brain. While individual calibrations are needed for new pulse sequences, the method of k-space determination can be used for any sequence of preparation pulses and readout gradient waveforms and should prove useful for other trajectories, including the rastered lines of echo-planar imaging. PMID- 9339452 TI - Estimating test-retest reliability in functional MR imaging. I: Statistical methodology. AB - A common problem in the analysis of functional magnetic resonance imaging (fMRI) data is quantifying the statistical reliability of an estimated activation map. While visual comparison of the classified active regions across replications of an experiment can sometimes by informative, it is typically difficult to draw firm conclusions by inspection; noise and complex patterns in the estimated map make it easy to be misled. Here, several statistical models, of increasing complexity, are developed, under which "test-retest" reliability can be meaningfully defined and quantified. The method yields global measures of reliability that apply uniformly to a specified set of brain voxels. The estimates of these reliability measures and their associated uncertainties under these models can be used to compare statistical methods, to set thresholds for detecting activation, and to optimize the number of images that need to be acquired during an experiment. PMID- 9339454 TI - Optimization of flip angle for T1 dependent contrast: a closed form solution. PMID- 9339453 TI - Estimating test-retest reliability in functional MR imaging. II: Application to motor and cognitive activation studies. AB - Functional magnetic resonance imaging (fMRI) using blood oxygenation contrast has rapidly spread into many application areas. In this paper, a new statistical model is used to evaluate the reliability of fMRI activation in a finger opposition motor paradigm for both within-session and between-session data and in a working memory paradigm for between-session data. A slice prescription procedure for between-session reproducibility is introduced. Estimates are made for the probabilities of correctly and falsely classifying voxels as active or inactive and receiver operator characteristic curves are generated. In the motor paradigm, estimated between-session reliability was found to be somewhat reduced relative to within-session reliability; however, this includes additional sources of variation and may not reflect intrinsically lower reliability. After matching false-positive classification probabilities, between-session reliability was found to be nearly identical for both motor and cognitive activation paradigms. PMID- 9339455 TI - Future research directions in laterality. AB - This paper considers the current conceptual state of research into neuropsychological laterality and considers some issues which might appropriately be considered for the forward development of the field. It considers the biological context which has been adopted for these studies and the psychological significance of performance asymmetries. A principal emphasis of the paper is the degree to which inferences, rather than direct methodological deductions, can be drawn from the research undertaken. The status of the dichotomies which have been proposed, the role of interhemispheric transfer, and stages of processing models are considered. The degree to which cerebral asymmetries may be inferred to reflect normal processes of the brain is questioned, and some prospects of the future discussed. PMID- 9339456 TI - Cognitive processing and self-report of lateral preference. AB - Our knowledge of the human brain has increased more during the past 40 years than at any other time in history. Of particular interest have been the findings of a correspondence between cognitive functions and individual structures of the brain. Similar from a gross anatomical point of view, the hemispheres of the brain have been shown to serve specialized cognitive functions. This work offers an overview of the cognitive aspects of cerebral lateralization as a context for considering this issue, followed by a review of specific self-report techniques in the appraisal of lateral preference. PMID- 9339458 TI - A pediatrician's view. Practice makes (more) perfect. PMID- 9339459 TI - Pediatric brain tumors. PMID- 9339457 TI - Unimanual performance as a measure of laterality. AB - The study of unimanual performance as a measure of laterality has ranged from simple concepts such as tests of handedness to highly complex conceptualizations interrelating anthropologic, cultural, genetic, and neurological aspects, including difference between unimanual performance in brain injured versus normal samples. This paper traces measures of unimanual performance through behavioral correlates of natural and unnatural sinistrality as related to cerebral organization, and concludes that unimanual performance represents a robust phenomenon with implications for understanding of neuropsychological correlates of behavior. PMID- 9339460 TI - The management of cerebral palsy: how can neurosurgery help? PMID- 9339461 TI - Craniosynostosis: diagnosis and management in the new millennium. PMID- 9339462 TI - Cerebrospinal fluid shunt problems in pediatric patients. PMID- 9339463 TI - Accidental head injury. PMID- 9339464 TI - The joint of Tn916 circular intermediates is a homoduplex in Enterococcus faecalis. AB - Tn916 is a 18-kb conjugative transposon originally identified in Enterococcus faecalis. The first step for Tn916 movement is its excision from the donor replicon with the formation of a nonreplicating covalently closed circular intermediate. Studies on formation of circular intermediates in Escherichia coli have shown that the joint between the Tn916 termini is a 6-bp heteroduplex formed by the two regions flanking the transposon before its excision (coupling sequences). In this work we studied the joint of Tn916 termini in circular intermediates formed in both E. coli and E. faecalis. Our strategy was to use direct sequencing of amplification products obtained from the joint region of single target molecules. In E. coli, 50% of circular intermediates contained a heteroduplex joint, while the remaining 50% displayed a homoduplex joint formed by one of the two coupling sequences. In E. faecalis, we could not demonstrate the presence of any heteroduplex joint. In this case 77.7% of the analyzed joints were formed by the left coupling sequence. PMID- 9339465 TI - Genetic analysis of regions involved in replication and cadmium resistance of the plasmid pND302 from Lactococcus lactis. AB - The 8.8-kb Lactococcus lactis plasmid pND302 encodes resistance to cadmium (CdR). Regions of pND302 involved in replication and CdR were subcloned and sequenced. The replication region is localized on a 1.5-kb region and consists of an open reading frame (repB) preceded by a noncoding AT-rich sequence (ori) which is highly homologous to lactococcal theta-type replicons. The CdR determinant is localized on a 2.9-kb region and encodes putative proteins similar to the Cd(2+) specific P-type efflux ATPase (CadA) and the transcriptional regulatory repressor (CadC) identified in Staphylococcus aureus, Bacillus firmus, and Listeria monocytogenes. Similar CdR determinants were also detected by PCR in other CdR plasmids isolated from different L. lactis strains. PMID- 9339466 TI - Minimum length of sequence homology required for in vivo cloning by homologous recombination in yeast. AB - With efficient homologous recombination in Saccharomyces cerevisiae, a rapid in vivo cloning technique has been available. Here we demonstrated that 30 bp of a homologous sequence at each end of a DNA fragment is sufficient to integrate the fragment into a linearized plasmid in yeast. To obtain a high yield of recombination transformants, however, more than 60 bp are desirable. Interestingly, we observed that 20 bp of homology at one end of a DNA fragment is sufficient for efficient recombination provided that the other end contains 80 bp of homologous sequence. Some applications, including high-throughput transferring of EST inserts to the yeast expression systems for the Human Genome Project, are discussed. PMID- 9339467 TI - Characterization of the replication and mobilization regions of the multiresistance Klebsiella pneumoniae plasmid pJHCMW1. AB - A 2.4-kb EcoRI fragment including the replication and origin of transfer regions of the Klebsiella pneumoniae multiresistance plasmid pJHCMW1 has been cloned and sequenced. The isolated replication region was sufficient for stable maintenance of the plasmid and shares homology with RNA-regulated replicons. Homology was highest with the replication region of the plasmid p15A. Incompatibility experiments, however, determined that pJHCMW1 is compatible with pACYC177, a plasmid harboring the p15A replicon. Differences in their RNA I nucleotide sequences may account for their compatibility. A mobilization origin was also found in the 2.4-kb EcoRI pJHCMW1 DNA fragment analyzed. Conjugation experiments showed that although non-self-transmissible, the recombinant clone including the 2.4-kb EcoRI pJHCMW1 fragment could be mobilized in the presence of the helper plasmid pRK2073. PMID- 9339468 TI - Identification of plasmids in the genus Marinococcus and complete nucleotide sequence of plasmid pPL1 from Marinococcus halophilus. AB - Several plasmids were detected in the Gram-positive halophilic eubacterium Marinococcus halophilus and in the related strain M52. The complete nucleotide sequence (3874 bp) of one of these plasmids, pPL1, was determined. Four major open reading frames were identified. Whereas orf3 and orf4 showed no sequence similarities to known proteins, rep displayed a high sequence similarity to replication proteins of rolling circle plasmids. Upstream of this ORF, a sequence resembling the double-strand origin was detected. A region probably constituting the single-strand origin was identified. The ORF mob showed sequence similarity with Mob proteins of rolling circle plasmids. The observed characteristics suggest that pPL1 replicates according to the rolling circle mechanism. PMID- 9339469 TI - Replication regions of two pairs of incompatible lactococcal theta-replicating plasmids. AB - Incompatibility tests were performed employing 12 replicons belonging to a family of homologous lactococcal theta-replicating plasmids. Two pairs of incompatible plasmids were found, namely, pFV1001 and pFV1201, and pJW565 and pFW094. The replicons of plasmids pFV1001, pFV1201, pJW565, pJW566, and pFW094 were sequenced. Alignments were made of the replicational origins (repA) and putative replication proteins (RepB) of these and 11 related plasmid sequences. Comparison of the alignments with the incompatibility data indicated that the incompatibility determinant could be contained within the 22-bp tandem repeats DRII and/or the inverted repeat IR1 in repA. In support, the incompatibility determinant of pJW563 was localized to a 743-bp fragment encompassing repA. A stretch of 13 amino acids of RepB was proposed to be responsible for the plasmid specific initiation of replication. This stretch is part of a domain containing features that are highly conserved within the proposed DNA binding regions of the initiation proteins from several well-characterized plasmids from Gram-negative bacteria, including pSC101, R6K, and mini-F. PMID- 9339470 TI - Cloning and characterization of a high-copy-number novel insertion sequence from chemolithotrophic Thiobacillus ferrooxidans. AB - Two distinct families of repetitive DNA elements (1.4 and 1.2 kb) were identified from S1 nuclease-treated genomic DNA of four strains of Thiobacillus ferrooxidans. The 1.4-kb fragment hybridized with IST2, an insertion sequence of T. ferrooxidans. The 1.2-kb fragment was cloned and sequenced. The sequence (IST445), 1219 bp in length, with features characteristic of an insertion element, has a terminal inverted repeat of 8 bp, which can be further extended to 23 or 48 bp with 9 and 26 mismatches, respectively. It displays 54.4% identity in 967 nucleotides of overlap with ISAE1 of Alcaligenes eutrophus. The IST445 contains three open reading frames which have codon usage almost similar to 56 different coding genes of T. ferrooxidans. In Southern blots of restricted genomic DNAs probed with IST445, each of the several strains of T. ferrooxidans gives a distinctive fingerprint. IST445 is present in the range of 10-20 copies per genome in the four strains studied. PMID- 9339471 TI - Electroporation-mediated transformation of Arcanobacterium (Actinomyces) pyogenes. AB - Plasmids derived from pNG2 or RSF1010 were introduced into strains of Arcanobacterium (Actinomyces) pyogenes by electroporation. Electroporation conditions were varied systematically to give a maximum electroporation frequency of 3.7 x 10(5) CFU/microgram DNA at 1.5 kV/cm and 246 omega, resulting in a time constant of approximately 10 ms. The A. pyogenes transformants expressed plasmid encoded resistance to chloramphenicol, erythromycin, kanamycin, and streptomycin. The source of incoming DNA affected the growth rate of transformants, but not the electroporation efficiency. This is the first report of genetic transformation of the veterinary pathogen A. pyogenes. PMID- 9339473 TI - Historical comments on regulation, public health, and cancer prevention. Remarks by the recipient upon his acceptance of the ISRTP 1996 International Achievement Award. PMID- 9339472 TI - The bioactive atelocollagen/hydroxyapatite composite as bone filler histological study on rats. PMID- 9339475 TI - Hormonal disruptors and male infertility: are men at serious risk? PMID- 9339474 TI - Vulnerability of the endocrine system to xenobiotic influence. AB - Natural sex hormones are most important factors guaranteeing the homeostasis of male and female sexual functions, including sexual differentiation and reproduction. Main target tissues include bone and skin, cardiovascular system, and possibly central nervous and immune systems. In medicine, synthetic hormonal substances with agonistic and antagonistic properties have been widely used for decades. Therapeutic benefit is the aim, and the many possibilities to interfere with normal or pathological hormonal situations are rather well understood. Synthetic hormonal agonists or (partial) antagonists may exhibit specific affinities to special receptors resulting in a spectrum of organotropies, or they may even induce opposite actions on different targets. Although not a new issue, environmental substances mimicking potentials of sex hormones have recently gained increased attention. There is not need to reinvent the wheel, since most (adverse) effects may be revealed with today's routine procedures used for testing medicinal substances, but some additional testing strategies should be included. Adverse effects of ecohormones may preferentially affect systems other than the human organism (assuming lower exposure and possibly lower susceptibility). Nevertheless, this survey is confined to possible alterations in the mammalian organism, since such effects are best understood from numerous experimental studies and clinical trials. PMID- 9339476 TI - Dose-response relationships for endocrine disruptors: what we know and what we don't know. PMID- 9339477 TI - A toxicologically based weight-of-evidence methodology for the relative ranking of chemicals of endocrine disruption potential. AB - A toxicologically based predictive scheme is presented for quantitatively ranking chemical agents with respect to their capacity to ensure endocrine disruption in target species based on short-term bioassays. Criteria providing the predictive framework include: (1) endocrine disruption as a multistage process, (2) phylogenetic considerations, (3) model system, and (4) estrogenic potency. Relative rankings were calculated for 15 environmentally relevant agents reported to have endocrine-disrupting effects. The relative ranking process offers a procedure for assessing the potential of endocrine disruption and for identifying data gaps for specific chemical agents. Although the current scheme is limited to "estrogenic" agents, it is anticipated that future refinements (e.g., incorporation of antiestrogenic potency data) will improve the system. PMID- 9339478 TI - Reflections on risk assessment of receptor-acting xenobiotics. PMID- 9339479 TI - Stepwise approaches for estimating the intakes of chemicals in food. AB - Estimating intakes of chemicals in food requires residue data, giving the concentration of the chemical in food, and consumption data for foods which could contain the chemical. Residue data are often restricted to only a few foods and even then may not be representative of the foods of interest due to limitations in the sampling or analytical procedures. Obtaining reliable consumption data can also be complex. People normally eat a wide range of different foods, which may each contain a range of concentrations of the chemical of interest. Both sets of data can be combined using one of several approaches. This paper reviews the nature and relevance of the factors affecting the collection of these two data sets and the procedures available for combining them to estimate consumer exposure. Considerable expertise and judgment are necessary to obtain an estimate of exposure that is robust enough for risk management purposes. PMID- 9339480 TI - Human exposure to endocrine-active chemicals: hazard assessment problems. AB - Industrial-derived endocrine disruptors or endocrine-active chemicals (EACs) have been identified and hypothesized to play a role in human disease. Most of the xeno-EACs which have been characterized bind to the estrogen receptor, aryl hydrocarbon receptor, or androgen receptor. Hazard and risk assessment of xeno EACs is complicated by several factors which include the following: (i) humans are exposed to relatively high levels of natural EACs compared to xeno-EACs; (ii) very little information on the effects of metabolism, serum, and intracellular binding proteins on target cell uptake of EACs is known; (iii) humans are exposed to EAC mixtures and their interactive effects may be additive, antagonistic, or synergistic and also response- and tissue-specific; and (iv) individual EACs may be agonists/antagonists for more than one endocrine response pathway. Scientific based hazard and risk assessment of both natural and xeno-EACs clearly requires more information on dietary intakes, target organ exposures, mechanisms of action, and interactive effects of mixtures. PMID- 9339482 TI - Endocrine disruptor risk characterization: an EPA perspective. AB - The characterization of risk to endocrine-disruptive agents may prove to be one of the greatest challenges that the risk assessment/regulatory community has ever faced. Why is this so? The endocrine system is actually many systems, having complex interactions and interdependencies. Normal endocrine function is often dependent on cyclical events, rather than steady-state. Timing is everything, as evidenced by significant differences in adverse outcome as a function of age and stage of development. Further, the consequences of concomitant exposures to endocrine-active substances in the diet or as therapeutic agents are poorly understood. So, how should risk characterization to these agents be approached? This presentation will include the description of current practices for addressing hormonally mediated cancer and noncancer effects and offer speculation on modifications to these approaches that might be necessary as our knowledge of this area increases. PMID- 9339481 TI - Development of a Tier I screening battery for detecting endocrine-active compounds (EACs). AB - One of the components of our research program is development of a mode-of-action screening battery to detect several different types of endocrine-active compounds (EACs). Our working hypothesis is that a comprehensive short-term in vivo/in vitro battery can be developed to identify endocrine toxicants using a collection of endpoints. The goals of this battery are that it be quick, cost effective, and predictive. The purpose of this battery is to identify potential EACs and to assess their potency in order to prioritize compounds for further study. Two in vivo screens (intact male and ovariectomized female rats) are being evaluated for their ability to detect several different types of endocrine activity. To validate this screen, 15 compounds with known endocrine activities are being used to evaluate a collection of different endpoints for their variability, stability over time, predictiveness, and dose dependency. These positive controls were chosen because they can modulate development, reproduction, or cancer. The advantage of an in vivo screen is that it utilizes a metabolically and physiologically intact system. The male in vivo battery will be used to assess several different types of endocrine activity, primarily by using a comprehensive hormonal battery. The female in vivo battery will be used to identify compounds which are either estrogenic/antiestrogenic or can alter the prolactin pathway. The in vitro portion of the screening battery consists of a yeast transactivation system (YTS). The YTS is being evaluated for its ability to identify compounds which are agonists or antagonists to the estrogen, androgen, or progesterone receptors. The expression of mammalian receptors in yeast allows for assessment of steroid-dependent transcriptional activators. The value of this system is that it can be used as a routine screen for compounds that interact with steroid receptors. Alterations in ligand binding to these receptors can be correlated with alterations in development via masculinization of females and/or feminization of males, decreases in reproductive success, or modulation of cancer incidence from in vivo tests. The in vivo and in vitro screens are designed to be run in parallel with built-in redundancy in order to reduce the probability of false-negative/ positive responses. PMID- 9339483 TI - Beyond screening: problems and prospects for risk characterization of endocrine disruptors. AB - Over time, the NAS's original concept of "risk characterization" has broadened to include not just explication of the risk inferences themselves, but a forthright discussion of their dependence on the many less-than-certain elements of the inferential process. Ideally, such characterization communicates the span of possible conclusions that emerge from alternative reasonable interpretations of the information at hand, together with considered judgment regarding the likelihood that each interpretation provides a good representation of the actual situation. Uncertainties regarding the magnitudes of parameters in the various equations used in quantitative risk assessment are increasingly characterized using statistical tools for analysis and propagation of variance through the calculations. Of potentially greater concern, however, is "model uncertainty"-the more fundamental uncertainty regarding the soundness of our conception of what biological phenomena are operating, how they interact, and how they can be represented in quantitative models. This is particularly true for assessment of potential endocrine disruptors, since much fundamental information is lacking and unanswered questions exist about the consequences and interpretation of empirically observable effects. An approach is to define a series of alternative models, each embodying a different conception, and using expert scientific judgment to weight the individual results. PMID- 9339484 TI - Assessing the risks of adverse endocrine-mediated effects: where to from here? AB - This article is based on the closing lecture (by J. Ashby) of the meeting held in Research Triangle Park on January 13-14, 1997-"Assessing the Risks of Adverse Endocrine-Mediated Effects." Several summaries of unpublished data from Zeneca CTL are presented to illustrate concerns over the present absence of agreed criteria for the assessment of endocrine toxicity data. Reference is also made to several points raised by others during the course of the meeting. It is concluded that the substantial challenges currently faced can be met only by strict adherence to the basic principles of good scientific practice, preeminent among which are the needs to confirm data and to respond to them in a cool and dispassionate manner. The wealth of expertise and enthusiasm being devoted to the task in hand, by both governmental and industrial interests, suggests that practical means by which to address the topic of the symposium will be forthcoming in the near future. However, moves to accelerate progress unduly-"to conduct science under the gun"-are already evident. Therefore, if external influences demand that short-cuts must be taken toward achievement of the goals set out in the Wingspread Statement (Colborn and Clement, 1992), such compromises should be clearly identified in order not to confuse the underlying progress that is already evident in this new branch of toxicology. PMID- 9339485 TI - Reproducibility of endocrine disruption data. PMID- 9339486 TI - Failure to confirm estrogenic activity for benzoic acid and clofibrate: implications for lists of endocrine-disrupting agents. AB - Earlier reports that benzoic acid is uterotrophic to the rat and mouse and that clofibrate is uterotrophic to the rat have not been confirmed. The studies reported here involved the use of a range of test protocols and dose levels, including the protocols/dose levels used by the original investigators. In addition, both chemicals were inactive in a human estrogen receptor (hER alpha) yeast estrogenicity assay. It is concluded that benzoic acid and clofibrate are not estrogenic in the assays used here. This conclusion has implications for the compilation of lists of endocrine-disrupting chemicals. PMID- 9339487 TI - Normal sexual development of rats exposed to butyl benzyl phthalate from conception to weaning. AB - Butyl benzyl phthalate (BBP) has been administered in drinking water (1000 micrograms/liter) to pregnant AP rats during gestation and lactation. The sexual development of the pups was then monitored until their termination at postnatal day 90 (pnd 90). Pups derived from animals exposed to diethylstilbestrol (DES) in drinking water (50 micrograms/liter) acted as a positive control group. Glass drinking bottles were employed following demonstration of absorption of BBP into plastic drinking bottles. Drinking water intake was monitored and the estimated average exposures of the dams was 182.6 micrograms/kg/day BBP and 8.6 micrograms/kg/ day DES. A total of 240 control pups, 204 BBP pups, and 64 DES pups were obtained for study. The sexual development of both genders of pups was monitored. The body weights of the DES pups were significantly reduced at birth, an effect that persisted until pnd 90. The body weights of the BBP pups were marginally increased at birth, but this difference resolved by pnd 90. DES affected the sexual development of the pups for all endopoints monitored anogenital (AG) distance on pnd 2; average day of vaginal opening and prepuce separation; uterus, testes, and accessory sex gland weight; and cauda epididymis sperm count and homogenization resistant testicular sperm count at pnd 90. BBP failed to affect any of these parameters, with the exception of a 1.1-day advance in the average day of vaginal opening and a small increase in male AG distance on pnd 2. These last two effects are related to the increased weight of the BBP pups. The incidence of FSH-containing cells in the pituitary gland of animals from each group was unaffected at pnd 90. The effects observed for the DES pups are consistent with the results of earlier studies by Sharpe et al. (Environ. Health Perspect. 103, 1136-1143, 1995). However, the absence of an effect of BBP administration on pup testis weight and testicular sperm count at pnd 90 is in contrast to reductions in these measurements reported earlier by Sharpe et al. (Environ. Health Perspect. 103, 1136-1143, 1995) for similarly derived BBP pups. PMID- 9339488 TI - A review of carbon disulfide exposure data and the association between carbon disulfide exposure and ischemic heart disease mortality. AB - Recent regulatory efforts have devoted attention to carbon disulfide (CS2) exposure and its potential effects on the cardiovascular system. To investigate the association between CS2 exposure and ischemic heart disease (IHD) mortality, the analysis presented here had the following objectives: (i) to review historical CS2 exposure data in the viscose rayon industry and identify trends and (ii) to use these historical data to suggest a standard mortality ratio (SMR) exposure relationship and a threshold level for occupational exposure to CS2, CS2 exposure data were extracted from published studies and used with the SMR versus exposure score relationship developed by Sweetnam et al. (Br. J. Ind. Med. 44, 220-227, 1987) to relate SMRs directly to exposure. Upper and lower bound exposure profiles were derived and used to identify exposure thresholds. For an IHD SMR equal to 100, the upper and lower bound exposures were 60 and 20 ppm, respectively. The analysis indicates that the risk of IHD mortality and its relationship to CS2 exposure is meaningful only for workers exposed to high level for many years. These high levels, which existed many years ago, are no longer found in the workplace. The results of this analysis suggest a safe regulatory exposure level for CS2 between 15 and 20 ppm. PMID- 9339489 TI - Marketing authorization of veterinary medicinal products in Poland. AB - Opening of the European Agency for the Evaluation of Medicinal Products (EMEA) in London (February 1, 1995) has begun harmonization of international requirements in veterinary drug legislation in the European Union (EU). The main objective of this paper is to introduce the principal rules of veterinary drug registration in Poland, a candidate to membership in the EU. In Poland the basic guideline governing quality, production, marketing, and inspection of medicinal products is "The Pharmaceutical Products, Medical Materials, Pharmacies, Wholesaler Outlets, and Pharmacy Inspection Act, dated October 10, 1991." The detailed rules are given in directives of the Minister of Health or the Minister of Agriculture in case of veterinary medicinal products. Since 1993 licensing of drugs in Poland has become a universal procedure for both human and veterinary products. The common Committee of Drug and Medical Materials Registration, which is under control of the Minister of Health and Social Welfare, ensures that the quality, safety (e.g., maximum residue limits and current FAO/WHO and EEC regulations for products used in food-producing animals are accepted), and efficacy are the criteria for the registration process. During 1994-1996 almost 300 veterinary products (more than 200 from the EU) received marketing authorizations in Poland. All these products were evaluated and registered on a similar basis to those in the European Union. PMID- 9339490 TI - Dioxin is a promoter blocker, a promoter, and a net anticarcinogen. AB - An ad hoc panel of dioxin experts convened by EPA's Science Advisory Board found the Agency's claim of cancer causation by dioxin unacceptable. This paper uses the very same two studies [Fingerhut (NIOSH) and Kociba] EPA relied upon to test the observations made in an unused third study [Bertazzi] to conclude that dioxin is (1) a promoter blocker of certain cancers, including all the cancers Agency scientists claimed dioxin promoted; (2) a promoter of some other cancers that EPA scientists failed to identify; and (3) a net anticarcinogen. Three independent total tumor reductions provide evidence of both cancer prevention and quantifiable risk reductions tied to specific dioxin levels. The author indirectly suggests a general cancer prevention treatment, even for cancers already initiated. PMID- 9339491 TI - External iliac artery endofibrosis in athletes. AB - Atherosclerosis and inflammatory arterial diseases are rare in young people. Since the early 1980s, an increasing incidence of iliac arterial stenosis in competition cyclists has been reported. Histological findings in these individuals are specific, with fibrosis of the intimal wall on histology and no atherosclerotic or inflammatory lesions. Clinical consequences of this arterial endofibrosis are usually described as an exercise-related subjective sensation of swollen thigh in one or both (15%) legs, with normal clinical and Doppler investigations at rest. Following maximal exercise, ankle-to-brachial systolic pressure index is lower than 0.5 in 85% of individuals with disease and is used as a key argument for diagnosis before deciding upon arteriography. Surgery (recalibrated saphenous grafts or angioplasty-endofibrosectomy) seems to be efficient to allow an early return to competition, but its long term results are still to be evaluated. The physiopathology of this disease and its possible relationship with atherosclerosis are unknown. PMID- 9339492 TI - Children's and adolescents' anaerobic performance during cycle ergometry. AB - Cycle ergometry studies originated in the early 1900s but it was not until the early 1970s that the first studies of children and anaerobic performance were established. Since that time, research into the anaerobic performance of children and adolescents has proliferated, mainly due to attempts by investigators to overcome methodological problems. Besides the increase in studies using the most popular anaerobic test, the friction-braked Wingate, other tests such as the force-velocity and isokinetic cycle ergometers are becoming more common. No matter how the data are standardised, there is unequivocal agreement that children's and adolescents' anaerobic power scores are lower than those of adults. Qualitative muscular differences are often cited for this disparity rather than differences in the quantity of muscle, but conclusive research is lacking in this area. Despite the ethical considerations involved in studies with children, cycle ergometry has aided researchers to assess external short term power output, mean power and fatigue. PMID- 9339493 TI - Cardiovascular responses to exercise in children. AB - The cardiovascular system of children responds to exercise differently than does that of an adult, although the mechanisms behind the differences are unclear. During dynamic exercise, it has been reported that heart rate (HR) response to the initiation of exercise is both faster and slower in children than adults. Furthermore, HR recovery has been reported to be faster in children. During submaximal steady state exercise, HR and total peripheral resistance are higher, while stroke volume [SV (ml)] and cardiac output [Q (L/min)] are lower in children at a given rate of work. At maximal exercise intensities HR is higher while SV and Q are lower in children than adults. Differences in cardiovascular responses to dynamic exercise between young boys and girls have also been reported. The majority of studies report that HR is lower and SV is higher in boys than girls at a given rate of work, although data to the contrary have been reported. These differences seem to be related to larger hearts in the boys. Further, the majority of the studies report that Q is similar in young boys and girls at a given rate of work. Few studies have reported differences between boys and girls at maximal intensities of exercise, and the results of those studies are inconsistent. Less is known about cardiovascular responses of children to static exercise compared with adults. A number of studies have reported that HR response to handgrip exercise is greater in children than adults, while others have reported no difference in this response. Even fewer studies have compared boys and girls in their cardiovascular response to static exercise and the results of these studies are also inconsistent. During prolonged exercise both children and adults exhibit cardiovascular drift (gradual increase in HR and decrease in SV). The direction and degree to which these changes differ between children and adults is unclear, with both greater and lesser responses being reported in children. Few studies have investigated differences in cardiovascular response to prolonged exercise between boys and girls. Those that have, report no difference between young boys and girls. PMID- 9339494 TI - Measurement of physical activity in children with particular reference to the use of heart rate and pedometry. AB - Understanding the progression of physical activity behaviour from childhood to adulthood requires a valid, reliable and practical method of assessing activity levels which is appropriate for use in large groups. The measurement of physical activity in large scale research projects requires a method which is low in cost, agreeable to the study volunteer and accurate. Self-report can be used to determine adult activity patterns, but children lack the cognitive ability to recall details about their activity patterns. Heart rate telemetry has been used to estimate daily activity in children as a sole criterion and to validate commercial accelerometers. However, heart rate is an indirect estimate of physical activity which makes assumptions based on the linear relationship between heart rate and oxygen uptake. It is sensitive to emotional stress and body position, and takes longer to reach resting levels after physical exertion compared with oxygen uptake. It also lags behind movement, particularly as children's physical activity is spasmodic or intermittent in nature. One alternative is the pedometer. Many early studies reported that the pedometer is inaccurate and unreliable in measuring distance or counting steps. While reasonably accurate at mid range speeds, the accuracy of the pedometer decreases in very slow walking or very fast walking or running. However, more recent studies have examined the efficacy of using pedometers to assess daily or weekly activity patterns as a whole, and these have produced more promising results. In this regard, the pedometer has a number of advantages. It is very cheap, objective and does not interfere with daily activities and is therefore appropriate for use in population studies. Commercial accelerometers with a time sampling mechanism offer further potential and could be used to provide a picture of the pattern of children's activity. As it has been observed that prolonged activity periods are not typically associated with childhood behaviour patterns, the use of a threshold value for 'aerobic' training stimulus is not appropriate as a cut-off value for physical activity. Instead, there is evidence to suggest that the total activity data measured by pedometers over limited periods of time may be more appropriate to assess how active children are. PMID- 9339496 TI - Analysis of medical images. PMID- 9339498 TI - The analysis of functional magnetic resonance images. AB - Functional magnetic resonance imaging (fMRI) is a powerful technique for measuring brain activity associated with the performance of different mental tasks. We briefly describe this technique and the images it produces and review methods of analysis that have been applied to fMRI data. Throughout the paper, we illustrate various points on an fMRI dataset. PMID- 9339497 TI - Medical image analysis with fuzzy models. AB - This paper updates several recent surveys on the use of fuzzy models for segmentation and edge detection in medical image data. Our survey is divided into methods based on supervised and unsupervised learning (that is, on whether there are or are not labelled data available for supervising the computations), and is organized first and foremost by groups (that we know of!) that are active in this area. Our review is aimed more towards 'who is doing it' rather than 'how good it is'. This is partially dictated by the fact that direct comparisons of supervised and unsupervised methods is somewhat akin to comparing apples and oranges. There is a further subdivision into methods for two- and three-dimensional data and/or problems. We do not cover methods based on neural-like networks or fuzzy reasoning systems. These topics are covered in a recently published companion survey by keller et al. PMID- 9339499 TI - The registration of multiple medical images acquired from a single subject: why, how, what next? AB - This paper reviews some of the recent techniques which have been used to register multiple images of the same patient. Image registration is a problem which has been receiving significant attention from the medical image processing community in recent years. A successful image registration can aid in patient diagnosis, treatment assessment, image guided interventions, surgery planning and surgery. At present the majority of work has focused on rigid body transformations of images. We shall discuss some of the approaches used and outline a key automatic method in detail. In order to allow image registration of parts of the body which do not remain rigid, either due to patient movement or a change in pathology, nonlinear deformation techniques are being developed. We shall talk of the history of these methods before explaining deformations using landmarks and a recent extension to allow the definition of rigid structures in such warps in more detail. Validation of these methods is of great importance and we shall discuss work which has already been carried out on this topic for rigid body registrations as well as ideas for the validation of deformation algorithms. PMID- 9339495 TI - Common hip injuries in sport. AB - As a major weight-bearing joint, normal hip function is fundamental to successful sporting participation. Not only is it important in running-, jumping- and kicking-based activities, it also contributes to the generation and transference of forces in upper limb-dominated activities. Injuries to the hip do not account for a large proportion of the sports physician's workload, but may result in significant morbidity. The wide variety of acute, subacute and chronic injuries, affecting both the joint and surrounding soft tissues, can prove a diagnostic dilemma. The predisposition and the types of injuries around the hip vary with the age of the athlete. The young child rarely sustains a significant injury but one should be aware of orthopaedic conditions common in this age group that may manifest themselves through exercise. The immature skeleton of the adolescent is relatively injury prone and the demands of sport often exceed the capacity of the growing musculoskeletal system. In adults and older athletes, a further spectrum of injury exists, along with the effects of aging tissues and the concerns of degenerative joint disease. Rational treatment is based on a clear diagnosis developed through sound knowledge and a thorough history and examination. For the sports physician, treatments are typically early physical therapy and structured, progressive rehabilitation programmes which are individualised to the needs of the athlete. The spectrum of hip injuries is reviewed with current recommended diagnoses and management. PMID- 9339501 TI - New advances in endovascular technology. AB - Future research on and development of intravascular stents, stent grafts, and other implantable devices will be aimed largely at improving their blood and tissue biocompatibility. Most of the modifications resulting from this research and development will alter the surface properties we currently understand: surface energy or "wetability," electrical surface charge, surface texture, and surface chemistry. However, as knowledge of the physical, chemical, and biological properties of bioprosthetic surface materials expands, new areas for exploration will develop, increasing the opportunities for improvement. PMID- 9339502 TI - Aortic aneurysm morphology for planning endovascular procedures. AB - Endovascular prosthesis repair of abdominal aortic aneurysms is based upon the development of low-profile devices that can be expediently deployed within the aneurysm, excluding it from intraluminal pressure. Many factors affect the treatment of patients in this manner, including the morphology of the proximal and distal fixation sites, the diameter and disease state of the access vessels, and the ability of the device to conform to the many anatomic variations of aneurysms. In addition, preliminary data suggest there is progressive shrinkage and morphologic change in the configuration of an aneurysm following exclusion, which not only affects the alignment of the device but may also influence its healing and stability. This paper reviews the morphologic parameters of aneurysms relevant to endovascular repair and describes the imaging technologies used to assess these parameters before, during, and after intervention. PMID- 9339500 TI - Statistical methods in computational anatomy. AB - This paper reviews recent developments by the Washington/Brown groups for the study of anatomical shape in the emerging new discipline of computational anatomy. Parametric representations of anatomical variation for computational anatomy are reviewed, restricted to the assumption of small deformations. The generation of covariance operators for probabilistic measures of anatomical variation on coordinatized submanifolds is formulated as an empirical procedure. Populations of brains are mapped to common coordinate systems, from which template coordinate systems are constructed which are closest to the population of anatomies in a minimum distance sense. Variation of several one-, two- and three-dimensional manifolds, i.e. sulci, surfaces and brain volumes are examined via Gaussian measures with mean and covariances estimated directly from maps of templates to targets. Methods are presented for estimating the covariances of vector fields from a family of empirically generated maps, posed as generalized spectrum estimation indexed over the submanifolds. Covariance estimation is made parametric, analogous to autoregressive modelling, by introducing small deformation linear operators for constraining the spectrum of the fields. PMID- 9339503 TI - Extracranial carotid angioplasty and stenting. Initial results and short-term follow-up. AB - Carotid percutaneous transluminal angioplasty, with or without stent implantation, is becoming another therapeutic option for carotid revascularization. To evaluate the feasibility and effectiveness of the technique, from October of 1995 to March of 1997, we performed 24 percutaneous transluminal angioplasty procedures in 22 patients with severe extracranial carotid artery stenosis. Three common carotid and 21 internal carotid arteries were treated, and 19 procedures included stent implantation using nonarticulated PALMAZ stents (P154 and P204). Twelve patients were asymptomatic and 10 patients were symptomatic; 2 of the symptomatic patients had complete obstruction of the internal carotid artery that was successfully recanalized. Technical and angiographic success was achieved in 23 of 24 procedures, with the carotid artery obstruction diminishing from 85.6% +/- 8.5% to 5.7% +/- 3.2% (P < 0.001). Average stenosis length was 12.5 +/- 3.1 mm, and mean time of carotid occlusion during balloon inflation was 11.5 +/- 2.5 seconds. Three patients experienced transitory seizures during the procedure prior to dilation, 1 patient had a minor stroke with complete recovery within 72 hours, and 1 patient had a major stroke and died 45 days after the procedure. Clinical follow-up was achieved in all patients (mean, 10.5 +/- 7.2 months) and angiographic follow-up in 16 patients (mean, 6.3 +/- 1.2 months). The results obtained in this initial experience provide adequate support to continue further evaluation of this new therapeutic strategy. PMID- 9339504 TI - Predictors of primary patency failure in Wallstent self-expanding endovascular prostheses for iliofemoral occlusive disease. AB - We studied the factors that affected the primary patency and the clinical and procedural success of WALLSTENTS (stents) that were used at our institution from 1 March 1994 to 30 October 1995 for the treatment of iliac and femoral artery occlusive disease. This prospective study comprised 63 patients with 82 lesions. Follow-up was performed for a mean duration of 18.7 months. Pre- and post procedural duplex ultrasonography, together with estimation of ankle-brachial index scores, was performed on all patients, and additional studies were performed at clinical follow-up if indicated. The technical success rate was 100%. Ankle-brachial index scores improved considerably from 0.52 +/- 0.21 before the procedure to 0.73 +/- 0.27 after the procedure. The significant predictors by univariate analysis of primary patency failure were: Fontaine class III or IV (P = 0.044); femoral location (P = 0.004); lesion length > 100 mm (P = 0.010); poor or moderate outflow (P = 0.026); and number of stents > or = 3 (P = 0.012). Cox regression analysis showed that > or = 3 stents (risk ratio = 5.61), poor or moderate outflow (risk ratio = 6.05), and femoral location (risk ratio = 5.18) were the significant predictors of primary patency failure. Femoral lesions required more stents than did iliac lesions (2.2 +/- 0.8 vs 1.3 +/- 0.5). Primary patency rates for iliac and femoral stents were 86% and 49%, respectively, at 12 months, and 82% and 41% at 24 months. PMID- 9339505 TI - What do we need to know to achieve durable endoluminal abdominal aortic aneurysm repair? AB - The exclusion of abdominal aortic aneurysms with endoluminal grafts is in its earliest stages, and the technology is in continuous transition. While results with 1st-generation devices have been somewhat discouraging in some cases, lessons learned from these initial attempts have led to considerable improvement in device design and deployment techniques. Lower-profile devices that are smaller and more flexible have made implantation less traumatic, and the incidence of endoleak formation has been reduced to 10% or less in some series. A modified percutaneous approach has also been introduced, and it may reduce the need for open exposure of the femoral artery in endoluminal graft procedures. Treating aneurysm expansion earlier, perhaps at 4 cm, may allow use of simpler, straight-tube prostheses and prevent problems associated with the use of larger, bifurcated endoluminal grafts. Numerous endoluminal graft designs are being tested, including both internally and externally covered prostheses. The success with a covered device may depend upon the type of material used and the extent to which it covers the endoluminal graft; fabric covering over a completely metal structure may allow a high degree of perioperative success and improvement in late outcome. The use of "hooks" to anchor or stabilize the endoluminal graft is also under study but is still controversial. The expense associated with endoluminal graft technology is currently high; therefore, it is likely that cost savings will be the result of shorter hospitalizations, little or no time spent in the intensive care unit, and fewer pre- and postoperative tests. PMID- 9339506 TI - Endoluminal grafting in the treatment of iliac and superficial femoral artery disease. AB - Treatment of iliac artery disease with stents has been generally successful; however, disease in the smaller arteries below the inguinal ligament has been more resistant to percutaneous intervention techniques. Ongoing research is evaluating the potential value of newer, more flexible stents as well as the use of covered endoluminal grafts to "reline" diseased arterial segments. It is possible that intimal hyperplasia may be reduced by covering the stent on one or both sides with a fabric such as polytetrafluoroethylene, yielding improvements in long-term patency. A number of device manufacturers have developed Investigational Device Exemption protocols with the Food and Drug Administration to allow randomized comparison of covered grafts and uncovered stents. The use of endoluminal grafts in the treatment of large aneurysms involving the common and internal iliac arteries and the origin of the external iliac artery is also under investigation; this application may prove advantageous, since operative intervention in these locations is often difficult. In addition, the endoluminal graft has been used to manage traumatic or iatrogenic rupture of an iliac artery, and the use of systems incorporating nitinol stents for an "internal" femoropopliteal bypass procedure is also being studied. Although aneurysmal disease in the superficial femoral artery is uncommon, the use of endoluminal grafts now makes it possible to treat these lesions percutaneously with an intraluminal approach; endoluminal graft exclusion of aneurysmal disease in the popliteal artery is also promising. PMID- 9339508 TI - Use of spiral computed tomographic angiography in monitoring abdominal aortic aneurysms after transfemoral endovascular repair. AB - Transfemoral endovascular repair of abdominal aortic aneurysms has proved to be technically feasible in a selected group of patients. However, long-term efficacy has not been proved. Graft performance after implantation can be monitored by a single imaging technique: spiral computed tomographic angiography. With this technique, the parameters for continuing clinical success of the procedure-graft patency, endoleaks, graft migration, attachment site diameter, attachment system failure, and aneurysm diameter-can be monitored. Only in selected cases will an additional imaging technique be necessary. PMID- 9339507 TI - Comparison of Wallgraft and Wallstent for treatment of complex iliac artery stenosis and occlusion. Preliminary results of a prospective randomized study. AB - We performed a prospective randomized study to compare the use of a bare metal stent (WALLSTENT Endoprosthesis) with use of a covered stent (WALLGRAFT Endoprosthesis)-both made by Schneider, Inc.; Minneapolis, Minn-for the treatment of complex iliac artery stenosis and occlusion. We report the preliminary results of a study performed at our institution from 1 February 1997 through 31 April 1997. The patient group was composed of 6 women and 4 men, with a mean age of 61.8 years (range, 47 to 73 years). Six WALLGRAFT endoprostheses (4 in the left iliac artery and 2 in the right) and 9 WALLSTENT endoprostheses (5 in the left iliac artery and 4 in the right) were implanted. The mean percent stenosis before treatment was similar in both groups (84.17% in the WALLGRAFT group and 82.14% in the WALLSTENT group). The post-treatment stenosis and peak systolic gradients were negligible or zero in both groups. The devices were safely deployed and technical success (< 30% residual stenosis) was achieved in both groups. The mean thigh-brachial index was similar in the 2 groups, both before treatment (0.65 in the WALLGRAFT group and 0.64 in the WALLSTENT group) and after treatment (1.12 in the WALLGRAFT group and 1.12 in the WALLSTENT group). Evaluation of clinical success revealed that symptoms of intermittent claudication improved markedly in 4 of 5 patients who received the WALLGRAFT Endoprosthesis. In the WALLSTENT group, 1 patient had symptomatic improvement, another had 1 limb improve and the other worsen, and the rest had no improvement. Clinical complications were observed in only 1 patient in the WALLGRAFT group and in 2 patients in the WALLSTENT group. These preliminary results indicate very good technical and early success at the 1-month follow-up with the use of the WALLGRAFT Endoprosthesis in complex iliac artery stenosis and occlusion. Despite these promising preliminary results, a longer follow-up study with a larger number of patients is needed to determine the benefits of the WALLGRAFT Endoprosthesis in patients with complex iliac artery stenosis or occlusion. PMID- 9339509 TI - Risk factor analysis among Egyptian patients who underwent coronary artery bypass surgery. AB - We conducted a retrospective review of Egyptian patients who underwent coronary artery bypass graft surgery at our institution between 1980 and 1995. We examined the prevalence of coronary artery disease risk factors and evaluated the early postoperative results. We then compared these results with the corresponding data in a subset of American patients who underwent coronary artery bypass grafting at our institution in 1993. There were 290 Egyptian patients: 275 men and 15 women. The mean age was 54.5 years (range, 30 to 70 years). Angina was present in 258 (89%) of the Egyptian patients; of these, 186 (72.1%) were in Canadian Cardiovascular Society class 3 or 4. Risk factor analysis revealed a high prevalence of hyperlipidemia (69.7%), cigarette-smoking (66.6%), family history of coronary artery disease (53.1%), hypertension (46.9%), obesity (46.2%), and diabetes mellitus (32.4%). Comparisons between the 2 groups showed that the risk factors, except for hypertension, were significantly higher in the Egyptian patients, despite the older age of the Americans (mean, 65.5 years; range, 22 to 88 years). The prevalence of triple-vessel disease was 86.6% in the Egyptian patients and 51.0% in the American patients (p < 0.001). The operative morbidity rates in the Egyptian patients were low: these included arrhythmias (13.8%), bleeding (13.4%), infection (7.6%), low cardiac output (3.4%), myocardial infarction (3.4%), and cerebrovascular accident (1.4%). The hospital mortality rate was 1.4% for the Egyptians and 1.7% for the Americans (NS). These results show that, despite the high prevalence of risk factors among Egyptian patients with coronary artery disease, coronary artery bypass grafting can be performed with low operative morbidity and mortality rates. PMID- 9339510 TI - The transseptal approach for mitral valve replacement revisited. AB - We describe our experience with the transseptal approach for mitral valve replacement, a technique that we applied especially in cases of 3rd and 4th operations wherein numerous adhesions made the usual left atrial approach difficult. We report 39 cases of mitral procedures in which we used 3 slightly different transseptal approaches, depending on the cardiac anatomy and the preferences of the surgeon. There were no complications associated with any of these approaches. Indeed they made the mitral valve procedure easier, because they enabled full exposure of the mitral valvular and subvalvular apparatus. We also propose the transseptal approach as a very safe and reproducible technique for use in patients with friable tissues, heavily calcified mitral valves, or small left atria- and in patients who must undergo combined tricuspid and mitral procedures. In this series, there were no conduction abnormalities secondary to the approach, nor were there any procedure-related deaths. PMID- 9339511 TI - The rare association of tetralogy of Fallot with hypertrophic cardiomyopathy. Report of 2 neonatal patients. AB - Although tetralogy of Fallot is commonly associated with other congenital heart defects, it is rarely found in conjunction with hypertrophic cardiomyopathy. We describe the cases of 2 neonates with this rare condition, both of whom required surgical intervention during infancy. Because hypertrophic cardiomyopathy is frequently familial, and tetralogy of Fallot is commonly found in patients diagnosed with chromosomal anomalies, we speculate about a possible genetic cause for this association. PMID- 9339512 TI - Takayasu's arteritis presenting as a mediastinal mass. AB - Patients with Takayasu's arteritis generally present with symptoms secondary to arterial insufficiency or with aneurysm formation. We report the unusual presentation and subsequent management of a patient with Takayasu's arteritis who developed symptoms secondary to an expanding mediastinal mass of unknown origin. PMID- 9339514 TI - Post-traumatic tricuspid valve insufficiency. 2 cases of delayed clinical manifestation. AB - We present 2 cases of tricuspid insufficiency following blunt chest trauma: 1 was diagnosed 5 months after the trauma and the other, 20 years after the trauma. In both patients, the tricuspid valve was replaced with a porcine bioprosthesis, because valve repair was not considered feasible. These cases emphasize the variability of clinical presentation of post-traumatic tricuspid valve insufficiency and indicate the need for close follow-up of patients after major thoracic trauma. PMID- 9339516 TI - Lutembacher's syndrome. PMID- 9339513 TI - Aortic valve replacement with a homovital valve. AB - "Homovital" allografts (viable homografts) are a good substitute for native aortic valves. A case of aortic valve replacement with a homovital aortic allograft is reported along with the results of immunologic investigations. Postoperatively, there was no clinical or echocardiographic evidence of valve dysfunction, and immunologic tests did not show evidence of graft versus host reaction. PMID- 9339515 TI - Anomalous origin of the right coronary artery from the pulmonary artery in association with a ventricular septal defect. AB - Origin of the right coronary artery from the pulmonary artery is a rare lesion occasionally found at angiography or autopsy. We report the rare preoperative diagnosis, in a child, of anomalous origin of the right coronary artery from the pulmonary artery, in association with a ventricular septal defect. The chest radiograph was normal, but auscultation revealed a continuous murmur at the left sternal border and electrocardiography showed right and left ventricular hypertrophy. A transthoracic echocardiogram depicted anomalous origin of the right coronary artery from the pulmonary artery. Color-flow Doppler echocardiography indicated possible right-coronary-artery-to-right-ventricle fistulae. Diagnosis was made by selective left coronary arteriography, which showed retrograde filling of the right coronary artery from collateral vessels. Selective left coronary arteriography depicted intercoronary flow, with no fistulae. Operative repair consisted of moving the proximal right coronary artery from its origin at the pulmonary trunk to the aorta. An associated procedure for correction of the ventricular septal defect was performed. The postoperative cardiac angiogram showed that the ventricular septal defect was closed and that flow through the right coronary artery was normal. Preoperative diagnosis of anomalous origin of the right coronary artery from the pulmonary artery is important, because this condition is surgically correctable. PMID- 9339517 TI - Osseous metaplasia and hematopoietic bone marrow in a calcified aortic valve. PMID- 9339519 TI - In situ anastomosis of the saphenous vein to the superficial femoral artery. PMID- 9339520 TI - Contentious issues in safety of diagnostic ultrasound. PMID- 9339518 TI - Congenital heart surgery in Houston. The early years. AB - During the 1950s and 1960s, major advances in medicine significantly influenced the development and application of surgery as treatment for congenital heart disease. The Texas Medical Center in Houston was at the forefront of these pioneering efforts and thus played an important role in the development of the art and science of congenital heart surgery. PMID- 9339521 TI - Cervical ultrasonography. PMID- 9339522 TI - Cervical funneling: sonographic criteria predictive of preterm delivery. AB - Our objective was to establish sonographic criteria that are predictive of preterm delivery in patients with internal os dilatation (funneling). The study population consisted of patients with cervical funneling identified on translabial or transvaginal ultrasound examination. Funnel length, functional length, percentage funneling and funnel width were evaluated for their predictive values for preterm delivery. In the 43 patients who met the study criteria, funneling was detected at a mean gestational age of 21.4 weeks (range 16-28). Twenty-three of 31 patients (74%), manually examined immediately following the ultrasound examination, had a closed cervix. Preterm delivery occurred in 42% of patients. Funnel length of > or = 16 mm, functional length of < or = 20 mm, funneling of > or = 40% and funnel width of > or = 14 mm correlated significantly with preterm delivery. Patients with funneling of < 25%, 25-50% and > 50% had preterm delivery rates of 17%, 29% and 79%, respectively. PMID- 9339523 TI - Dandy-Walker malformation diagnosed before 21 weeks of gestation: associated malformations and chromosomal abnormalities. AB - This study examined rates of concomitant structural and chromosomal abnormalities in 14 fetuses with a diagnosis of Dandy-Walker malformation or Dandy-Walker variant before 21 weeks' gestational age, compared to 14 fetuses with a diagnosis of Dandy-Walker malformation or variant between 21 weeks' gestation and delivery. A total of 24 fetuses had Dandy-Walker malformation and four had Dandy-Walker variant. Eight of the fetuses with the malformation had ventriculomegaly: one of the fetuses with early diagnosis and seven with later diagnosis (p = 0.027). None of the fetuses with Dandy-Walker variant had ventriculomegaly. The overall prevalence of concomitant structural abnormalities was 13/28; 8/14 for fetuses with early prenatal diagnosis and 5/14 for fetuses with late prenatal diagnosis of the malformation or the variant. Chromosomal abnormality rates were significantly higher among fetuses with early prenatal diagnosis (7/14) than among those with later prenatal diagnosis (1/14; p = 0.032). Abnormal karyotypes were more prevalent among fetuses without ventriculomegaly (7/20), compared to fetuses with ventriculomegaly (1/8). We conclude that fetuses with an antenatal diagnosis of Dandy-Walker malformation or Dandy-Walker variant before 21 weeks' gestational age have worse prognosis than fetuses with a later prenatal diagnosis of the same defect. PMID- 9339524 TI - Isolated choroid plexus cysts and association with fetal aneuploidy in an unselected population. AB - We sought to determine the relationship between an isolated choroid plexus cyst diagnosed antenatally and fetal aneuploidy in an unselected population at a district general hospital. Over a 5-year period all women attending for a detailed anomaly scan at 18-20 weeks' gestation were screened for evidence of a fetal choroid plexus cyst. All cases of choroid plexus cyst were recorded prospectively. The size, position and number of the cysts were noted and associated abnormalities seen on ultrasound were also recorded. Cases of choroid plexus cyst associated with fetal aneuploidy were noted. A total of 13,690 women were screened, and 84 cases of choroid plexus cyst were identified (0.6%). Of these, 41% underwent prenatal karyotyping by amniocentesis; 78 of 84 cases (93%) were isolated. Six had other markers for aneuploidy, and three of these fetuses had trisomy 18. All cases of isolated choroid plexus cyst resulted in chromosomally normal neonates. This was confirmed by either normal antenatal karyotype or postnatal examination by the pediatricians. The size, position and number of cysts did not appear to influence the risk of aneuploidy. We conclude that the risk of aneuploidy for a case of isolated choroid plexus cyst in an unselected population appears to be very low, and in this series was 0%. In this setting, we suggest detailed ultrasound examination is essential, rather than routine karyotyping. PMID- 9339525 TI - New charts for ultrasound dating of pregnancy. AB - We describe new charts and tables for dating pregnancies derived from data collected in a study designed to enable the construction of these and charts of fetal size. This was a prospective study in which 663 fetuses seen in the routine ultrasound clinic in a London teaching hospital were scanned once only for the purpose of this study. We discuss the statistical methodology in detail and compare the new charts with other published charts. We suggest that the differences seen may be due to variation in study methodology. PMID- 9339526 TI - Transvaginal ultrasonographic characterization of ovarian masses: comparison of five scoring systems in a multicenter study. AB - The aim of this work was to test and compare the accuracy of five different morphological scoring systems to identify malignant ovarian masses in a prospective multicenter study. Four of the systems had previously been reported by Granberg, Sassone, De Priest and Lerner and the fifth is newly developed. A total of 330 ovarian neoplasms were collected in three different centers, which adopted the same diagnostic procedures. Of these, 261 masses were benign (mean diameter 50 +/- 26 mm) and 69 were malignant (mean diameter 69 +/- 33 mm) (prevalence 21%). The area under the receiver operating characteristic (ROC) curve for the multicenter score was 0.84. This was significantly better than the areas of the other four scores which ranged from 0.72 to 0.75. The cut-off levels derived from the five ROC curves achieved a sensitivity that ranged from 74% (Sassone score) to 88% (De Priest score > or = 5), and a specificity from 40% (De Priest) to 67% (multicenter); the highest positive predictive value was 41% (multicenter). With a cut-off level of 9, the accuracy of the multicenter score was significantly better than the scores of Granberg and De Priest (McNemar's test p < 0.0001). Similar results were obtained in 207 ovarian masses of < or = 5 cm in mean diameter, and when 19 borderline and 11 stage 1 cancers only were considered. For the clinical purposes of a screening test we also checked a possible cut-off level of > or = 8, which increased the sensitivity to 93% with a drop of specificity to 56%. With the use of the same criteria for the scores of the different authors, the following values were obtained for sensitivity: 96%, 81%, 93% and 90%; and for specificity: 23%, 56%, 28% and 49%. The multicenter score performed well at distinguishing malignant from benign lesions, and was better than the other four traditional scores, for both large and small masses. This was mainly due to the introduction of two criteria that allowed correction for typical dermoids and endohemorrhagic corpora lutea. A completely reliable differentiation of benign from malignant masses cannot be obtained by sonographic imaging alone. PMID- 9339527 TI - Reproducibility of transvaginal Doppler velocimetry measurements in the uterine arteries of postmenopausal women. AB - Intraobserver and interobserver reproducibility of transvaginal Doppler velocimetry measurements in uterine arteries were assessed by two observers in 20 postmenopausal women. In addition, the agreement between the observers regarding the detection of an ovary as well as the presence or absence of intraovarian arterial blood flow was documented. Pulsatility index (PI), peak systolic velocity (PSV) and time-averaged maximum velocity (TAMV) measurements in uterine arteries were made twice by the same investigator (IJ). Thereafter, the same measurements were made by the second investigator (AT). The agreement in categorical data was studied by using 2 x 2 tables and Cohen's kappa-coefficient. Reproducibility of the Doppler measurements was analyzed by using coefficient of variation, repeatability coefficient, intraclass correlation coefficient, mean differences and limits of agreement. In addition, the 95% confidence interval was calculated as appropriate. There was agreement of 95-100% between the observers with respect to the detection of an ovary in transvaginal scanning. Agreement regarding the presence or absence of intraovarian arterial blood flow, however, was only 75-81%. Intraobserver repeatability was very good, the intraclass correlation coefficient being 0.98-0.99 and the coefficient of variation 6% in PI measurements. The PSV measurements showed an intraclass correlation coefficient of 0.94-0.96 and a coefficient of variation of 10-12%. The intraclass correlation coefficient for TAMV measurements was 0.94 and the coefficient of variation varied from 12 to 16%. Interobserver agreement was good. There was no bias between the observers' measurements. The intraclass correlation coefficient for PI measurements was between 0.93 and 0.95, and the coefficient of variation was 11%. For PSV measurements, the intraclass correlation coefficient was 0.79-0.80 and the coefficient of variation was 18-29%. TAMV measurements showed an intraclass correlation coefficient of 0.83-0.84 and a coefficient of variation of 19-29%. When the limits of agreement for Doppler velocimetry measurements are considered, we expect the two observers to give PI measurements that differ by less than 0.7, with any discrepancy being equally likely in either direction. In conclusion, based on the high intraclass correlation coefficients, the intraobserver repeatability of all Doppler parameters was very good, the most reliable measurement being the PI. Variation in PSV and TAMV measurements increased considerably, however, when Doppler velocimetry was carried out by two investigators instead of one. PMID- 9339528 TI - Color Doppler energy imaging in the diagnosis of fetal intracranial hemorrhage in the second trimester. AB - Unlike conventional color Doppler imaging; color Doppler energy (or power Doppler) displays the intensity of the returning Doppler signal, is less dependent on the orientation of the blood vessel, and is therefore better able to detect low blood velocities. For these reasons it could be useful in some investigations which are difficult to perform, such as transvaginal evaluation of fetal brain vessels. We report a case of a fetal intracranial hyperechoic lesion detected at 26 weeks by transabdominal sonography in a severely growth-retarded fetus. There was absence of diastolic flow in the umbilical artery and low impedance to diastolic flow in the middle cerebral arteries. The fetus was further investigated by transvaginal sonography for the evaluation of the nature and localization of the lesion and an intraventricular hemorrhage in the right brain parenchyma with disorganized supratentorial brain structure was observed. As color Doppler energy imaging is more sensitive to slow flow, it was more reliable than conventional Doppler imaging in confirming the absence of flow within and around the hyperechoic lesion in contrast to the normal vascularity of the contralateral ventricular system. After informed parental counselling, the mother, for psychological reasons, asked to be delivered by Cesarean section. The fetus died 24 h after birth. The autopsy corroborated the ultrasonographic diagnosis. This case report confirms the accuracy of transvaginal ultrasonography in the diagnosis of intracranial hemorrhage and suggests a specific role for color Doppler energy imaging. PMID- 9339529 TI - Mature teratoma arising from an intra-abdominal undescended testis presenting as a fetal abdominal mass. AB - Teratoma of the intra-abdominal testis is a rare finding in infants. We describe the case of a full-term newborn treated for a calcified abdominal mass which was observed unexpectedly on prenatal sonography. An undescended right testis was also noted. During laparotomy, a twisted retroperitoneal tumor was found just above the right deep inguinal ring. Histological analysis revealed a mature teratoma of the intra-abdominal right testis. The tumor was removed and there was no recurrence at follow-up 1 year later. A teratoma should be considered in cases of fetal abdominal mass, especially when the testes cannot be detected in the scrotum by the 8th month. Prenatal sonographic diagnosis might be possible. PMID- 9339530 TI - Antenatal intestinal vascular accident with subsequent small bowel atresia: case report. AB - A woman was referred at 25 weeks' gestation with decreased fetal movements. Ultrasound revealed a large solid fetal abdominal mass and gross fetal ascites. Amniocentesis and viral titers were normal. On subsequent ultrasound examinations, the mass and ascites slowly disappeared, but a small bowel obstruction developed. Spontaneous labor occurred at 35 weeks and the child was born with a distended abdomen. At laparotomy there was type 3 jejunal atresia, indicating that the fetal mass and ascites were secondary to this antenatal small bowel ischemia. PMID- 9339531 TI - Color Doppler sonographic features of uterine arteriovenous malformations: report of two cases. AB - We describe the color Doppler sonographic features of uterine arteriovenous malformations in two cases. In both cases color Doppler imaging demonstrated hypervascularity throughout the arteriovenous malformation. The dominance of pale shades during both systole and diastole represented low-impedance, high-velocity flow within the lesion and a colored mosaic pattern representing turbulent flow was noted. Spectral analysis of the vessels within the lesion confirmed high velocity flow during both systole and diastole, and a low resistance index. The spectral waveform trace also showed spectral broadening consistent with turbulence and the spectral envelope was irregular. These findings indicated the presence of numerous arteriovenous shunts and marked turbulence within the arteriovenous malformation. Spectral analysis of the venous flow revealed high flow velocities and systolic velocity peaks similar to an arterial pattern. The uterine artery velocity waveforms were characterized by high flow velocity and a low resistance index. The diagnosis of uterine arteriovenous malformation was confirmed by histological examination in both cases. The findings of these two cases suggest that color Doppler sonography may play an important role in the diagnosis of uterine arteriovenous malformations. PMID- 9339532 TI - Uterine sarcoma: diagnosis with multiparameter sonographic analysis. PMID- 9339533 TI - Nuchal translucency measurement and second-trimester biochemical screening for Down's syndrome. PMID- 9339534 TI - How do we find the moles most at risk for persistent disease? PMID- 9339535 TI - Absence of significant hemodynamic changes in the fetus following maternal betamethasone administration. PMID- 9339536 TI - Effects of emotion-related surface similarity in analogical problem solving. AB - In analogical problem solving, a source problem with a known solution is used to solve a target problem. The present study deals with one possible condition influencing the search for possible source problems (i.e., with similarities between source and target problems in the emotional connotation of the problem cover stories). Subjects were given six source problems-distractors as well as target-relevant problems-that were varied with respect to the emotional valence of the cover stories. Then one group of subjects (n = 32) was given a pleasant target problem, while the other group (n = 31) received an unpleasant target problem. Except for emotional valence the two target problems were identical. Subjects preferred those target-relevant source problems that were emotionally congruent with the target problem. The findings are interpreted within network theories of long-term memory, introducing emotional markers or emotion-concept nodes to represent the emotional connotation of the represented units. By controlling subjects' mood change after reading the (un)pleasant target-problem cover story, it could be ruled out that the observed results were due to a mood congruity effect of the type described by Bower (1981). PMID- 9339537 TI - Size scaling and spatial factors in visual attention. AB - This study investigates the effect of the visibility of near and far letter distractors on target processing by scaling the size of the distractor letters to compensate for changes in resolution across the visual field. In Experiment 1, scaled and unscaled distractors were presented at varied stimulus onset asynchronies. Results showed that scaling the size of distractor letters in relation to their distance from the target was effective in producing strong compatibility effects. Scaled distractors presented prior to, or simultaneously with, the target were found to interfere with target processing whether they were near or far from the target. Experiment 2 used scaled distractors and varied the presentation location and the amount of time for processing prior to the presentation of the target. Compatibility effects were found to vary by location and by the exposure duration of the distractor. The finding of distance effects at far locations supports a space-based visual-attention mechanism with a wide attention beam (Steinman, Steinman, & Lehmkuhle, 1995). PMID- 9339538 TI - Verbal-overshadowing effect: evidence for a general shift in processing. AB - Two experiments investigate the nature of the verbal-overshadowing effect-the finding that recognition performance for certain stimuli is impaired if it is described verbally (Schooler & Engstler-Schooler, 1990). Impairment on a face recognition task was found, although participants produced not a verbal description of the target but, instead, a description of another object (a car) presented in the study phase. These results support the idea that the verbal overshadowing effect reflects a general shift in the processes involved in face recognition rather than a specific impairment for the described stimulus. Results also support the notion that the impairing effect of verbalization is unique to certain types of stimuli; verbalization impaired recognition of a face but not of a car. PMID- 9339539 TI - Automaticity and consciousness: is perceiving the word necessary for reading it? AB - Definitions of automaticity imply insensitivity of the Stroop effect to conscious perceiving of the word. Subjects in the Stroop task reported the meaning of the stimulus word (in addition to its color) in 7% of the trials. The magnitude of the Stroop effect in these trials was correlated with subjects' ability to report the stimulus meaning. Furthermore, the effect was absent when subjects failed to report the stimulus meaning. These findings challenge the assumption that automatic processing is unconscious. A distinction between automatic and non automatic processing in terms of modes of consciousness is discussed. PMID- 9339540 TI - Effects of the nuclear genome on selective transmission of mitochondrial DNA in Drosophila. AB - In mitochondrial DNA (mtDNA) heteroplasmy in Drosophila, we previously reported that mtDNA was selectively transmitted depending on temperature (Matsuura et al., 1991). To investigate the effects of nuclear genome on the temperature-dependent transmission, two sets of heteroplasmy were constructed by germ-plasm transplantation, and changes in the relative proportion of two types of mtDNA were examined at 19 degrees C and 25 degrees C. In heteroplasmy possessing D. melanogaster and D. mauritiana mtDNA, two different nuclear genomes of D. melanogaster were examined after reciprocally substituting the nuclear genomes. In heteroplasmy possessing D. simulans and D. mauritiana mtDNA, nuclear genomes of D. melanogaster, D. simulans and D. mauritiana were used. For each set of mtDNA combination, the modes of temperature-dependent transmission of mtDNA differed according to the nuclear genome used. From these and our previous results (Matsuura et al., 1991; Tsujimoto et al., 1991), it is clear that the temperature-dependent transmission of mtDNA is affected by nuclear genome. This suggests that the nuclear genome is involved in determining the temperature dependency of mtDNA transmission. PMID- 9339541 TI - RIRE1, a retrotransposon from wild rice Oryza australiensis. AB - RIRE1 is a retrotransposon present in wild rice Oryza australiensis in an extraordinary number of copies, and only a portion of the LTR sequence has been determined previously. Here, we isolated and sequenced DNA segments of various portions of RIRE1, revealing that the sequences of LTR and the internal region were 1523 and 5277 bp in length, respectively. The internal region shows homology with the pol region in copia, a Drosophila retrotransposon, indicating that RIRE1 is a copia-like retrotransposon. The internal region of RIRE1 contained an open reading frame coding for genes, gag, pro, int, rt and rh, like copia and retroelements related to it. A clone screened from a library of the O. australiensis genomic DNA contained solo LTR, which was flanked by direct repeats of a 5-bp sequence. This suggests that RIRE1 generates a duplication of the target sequence of 5 bp upon retroposition. We observed that many RIRE1 members were nested by another RIRE1 member. This indicates that these RIRE1 members have received another RIRE1 to make an extraordinary number of copies in the O. australiensis genome without giving a deleterious effect on the growth of rice cells. PMID- 9339542 TI - A new Arabidopsis mutant induced by ion beams affects flavonoid synthesis with spotted pigmentation in testa. AB - A new stable mutant of Arabidopsis thaliana with a spotted pigment in the seed coat, named anthocyanin spotted testa (ast), was induced by carbon ion irradiation. The spotted pigmentation of ast mutant was observed in immature seeds from 1-2 days after flowering (DAF), at the integument of the ovule, and spread as the seed coat formed. Anthocyanin accumulation was about 6 times higher in ast mutant than in the wild-type at 6 DAF of the immature seeds, but was almost the same in mature dry seeds. A higher anthocyanin accumulation was not observed in the seedlings, leaves or floral buds of ast mutant compared with the wild-type, which suggests that a high accumulation of anthocyanins is specific to the seed coat of the immature ast seeds. Reciprocal crosses between ast mutant and the wild-type indicated that ast is a single recessive gene mutation and segregates as a delayed inheritance. The results of crossing with tt7 and ttg mutants also confirmed that the AST gene is probably a regulatory locus that controls flavonoid biosynthesis. A mapping analysis revealed that the gene is located on chromosome I and is closely linked to the SSLP DNA marker nga280 with a distance of 3.2 cM. AST has been registered as a new mutant of Arabidopsis. PMID- 9339543 TI - The Escherichia coli ldcC gene encodes another lysine decarboxylase, probably a constitutive enzyme. AB - A gene (designated ldcC) mapped at 4.6 min on the Escherichia coli chromosome codes for a protein of 713 amino acids (aa) that shows strong similarities in both size and amino-acid sequence (69% identical residues and 85% conserved residues) to lysine decarboxylase (LDC) from E. coli (CadA, acid-inducible LDC, 715 aa) or from Hafnia alvei (739 aa). A pUC18 derivative carrying the ldcC gene conferred high LDC activities on an E. coli strain devoid of the functional cadA gene, even when the bacteria were grown under non-inducing conditions at physiological pH. Thus, the gene encodes another lysine decarboxylase, probably a constitutively expressed enzyme, the presence of which was suggested from the previous observations that low LDC activities were detectable in cadA- mutant and non-induced wild-type cells. PMID- 9339544 TI - Cloning and nucleotide sequence of the putative polyketide synthase genes for pradimicin biosynthesis from Actinomadura hibisca. AB - We cloned the putative polyketide synthase genes (pms genes) for pradimicin A biosynthesis from Actinomadura hibisca using an oligonucleotide probe designed on the basis of conserved amino acid sequences of other polyketide synthases (PKSs). By DNA sequencing of an 8.2-kb SacI fragment that hybridized with the oligonucleotide probe, 11 open reading frames (ORFs) were found. All of the ORFs except for ORF10 were predicted to be translated in the same direction. Each of the deduced ORFs has significant sequence similarity to the protein responsible for polyketide biosynthesis or spore pigmentation. In particular, ORF1, ORF2, and ORF3 were 50-70% identical with genes coding for PKSs for actinorhodin biosynthesis. Specific DNA regions similar in sequence to pms genes were found with genomic Southern hybridization in all of the pradimicin producers examined, but were not found in pradimicin nonproducers, suggesting that the genes cloned in this study encode polyketide synthase for pradimicin biosynthesis. PMID- 9339545 TI - Changes in blood coagulation, platelet aggregation, and lipid metabolism in rats given lipids containing docosahexaenoic acid. AB - In order to identify an adequate intake level of docosahexaenoic acid (DHA), changes in various parameters related to health benefits were studied in rats fed on diets containing 10% test lipids at different n-3(DHA)/n-6 ratios for two weeks. An evaluation of the critical level of the dietary n-3/n-6 ratio which had a significant effect on the parameters of several tissues indicated that the response to the dietary ratios differed according to the parameter, the variation in ratio ranging approximately from 0.20 to 1.77 with either a positive or negative effect on the health benefit. These results suggest that a suitable intake level of DHA would be within this range. In view of safety, however, the critical level for the dietary n-3/n-6 ratio may be around 0.56, as shown by a detailed analysis on the lower limit level of the harmful parameters. We thus propose that the dietary intake of DHA should not be more than 0.56 in terms of the n-3/n-6 ratio. PMID- 9339546 TI - Molecular cloning and nucleotide sequence of the arginase gene of Bacillus brevis TT02-8 and its expression in Escherichia coli. AB - The gene from Bacillus brevis TT02-8 encoding arginase was cloned into Escherichia coli, and its nucleotide sequence was identified. The nucleotide sequence contained an open reading frame that encoded a polypeptide of 298 amino acid residues with a predicted molecular weight of 31,891, which was consistent with that previously calculated for arginase purified from this bacterium. Comparison of the deduced amino acid sequence of the B. brevis TT02-8 arginase with that of the prokaryotic and eukaryotic arginases of Bacillus caldovelox, Bacillus subtilis, Agrobacterium Ti plasmid C58, Saccharomyces cerevisiae, Coccidioides immitis, Xenopus laevis, Rana catesbeiana, rat liver, and human liver, showed 33-66% of the sequences to be similar; there were several highly conserved regions. Arginase activity was detected in Escherichia coli cells transformed with an expression plasmid of the cloned arginase gene. PMID- 9339547 TI - A conjugative linear plasmid in Streptomyces laurentii ATCC31255. AB - Plasmid pSLL of Streptomyces laurentii ATCC31255 (wild-type strain P0) is a 93 kilobase linear DNA plasmid that carries a protein bound to each 5' end of the DNA. It was self-transmitted to the pSLL-cured strain by conjugation in solid culture. The pSLL-cured strain carried a circular plasmid, pSLS, and showed a marked decrease in spore formation and thiostrepton productivity, owing to the pSLS. However, by retransmission of pSLL, these things reverted to levels seen in strain P0. Thus, plasmid pSLL suppressed the injurious effects of pSLS on the host mycelia. PMID- 9339548 TI - The effects of corn peptide ingestion on facilitating alcohol metabolism in healthy men. AB - We prepared corn peptide (CP), a vegetable oligopeptide and tried to discover the effects of its ingestion on facilitating alcohol metabolism in healthy adult men. Ten healthy male volunteers ingested 5 g of CP, wheat peptide (WP), pea peptide (PP), alanine, or leucine 30 min before alcohol intake at a dose of 0.5 g/kg, and blood ethanol and plasma amino acid concentrations were measured during a 2-h observation period after alcohol intake. In subjects who ingested CP, the blood ethanol level was lower than that in the WP, alanine and leucine ingestion groups, but did not decrease as compared to the control when they ingested PP. Similarly there was a difference in the blood ethanol level between alanine and leucine ingestion groups, and leucine ingestion was more effective than alanine against the reduction of the increase in blood ethanol level. On the other hand, there was no significant difference in the plasma concentrations of individual amino acids except alanine, leucine, or lysine after alcohol intake among experimental groups as compared to the control. CP ingestion significantly elevated plasma alanine and leucine rather than other groups during a 2-h observation period. These results suggested that CP may have the effect on the reduction of increase in blood ethanol level after alcohol intake by the marked elevation of plasma alanine and leucine, especially leucine, but neither by the delay of ethanol release from the stomach nor malabsorption of ethanol in the gastrointestinal tracts. PMID- 9339549 TI - Purification and characterization of ferredoxin-sulfite reductase from turnip (Brassica rapa) leaves and comparison of properties with ferredoxin-sulfite reductase from turnip roots. AB - Ferredoxin-sulfite reductase (Fd-SiR) [hydrogen-sulfide: ferredoxin oxidoreductase, EC 1.8.7.1] from turnip leaves (SiR-L) has been purified to homogeneity and its enzymatic properties compared with that from turnip roots (SiR-R). Each enzyme had a molecular mass of 64.5 +/- 0.5 kDa by SDS-PAGE and an isoelectric point of 5.15 +/- 0.05. Although each had a pH optimum around 7.8 with the same effects of inhibitors, SiR-L had higher heat stability at 60 degrees C than SiR-R. Moreover, SiR-R had a lower K(m) and a higher specificity constant (kcat/K(m)) for turnip leaf ferredoxin than SiR-L. The N-terminal amino acid sequence of SiR-L was different from that of SiR-R. The results of amino acid analysis and peptide mapping suggested that SiR-L and SiR-R have different primary structures. PMID- 9339550 TI - Subcellular location of polyphenol oxidase in apples. AB - The location of polyphenol oxidase (PPO) in cells of apple fruit was examined by immunohistochemistry and subcellular fractionation. In mature apple fruits, where vacuoles occupy most of the cells, PPO was detected immunochemically near the cell walls with use of anti-apple PPO antibodies. In cells of immature fruits and tissue culture, PPO was detected in organelles other than the vacuoles, probably in plastids. The plastid fraction was purified by density gradient ultracentrifugation, and the activities of PPO and marker enzymes of plastids were the highest in the plastids. Most apple PPO was in plastids, as are other plant PPOs, and some of the protein was solubilized and proteolyzed during ripening and storage. PMID- 9339551 TI - Expression and characterization of sucrose synthase from mung bean seedlings in Escherichia coli. AB - The cDNA fragment coding for mung bean (Vigna radiata Wilczek) sucrose synthase was introduced into the expression vector pET-20b resulting in the construction of plasmid pEB-01. After transformation of Escherichia coli strain BL21(DE3) cells by pEB-01 and induction with isopropyl thio-beta-galactoside, high level expression of the recombinant enzyme was obtained. The enzyme had a tetrameric form that conserved the activity of sucrose synthase. Although the Km and Vmax of the recombinant enzyme acting on either UDP-glucose or fructose were very close to those of the native enzyme isolated from mung bean seedlings, the Km for sucrose was higher by a factor of 10 for the recombinant enzyme. This suggests that the recombinant sucrose synthase has a tendency to synthesize sucrose, although the native enzyme catalyzes a freely reversible reaction. PMID- 9339552 TI - Inhibition of collagenases from mouse lung carcinoma cells by green tea catechins and black tea theaflavins. AB - Theaflavin and theaflavin digallate, which are components of black tea were examined by in vitro invasion assay with mouse Lewis lung carcinoma LL2-Lu3 cells, which are highly metastatic. The compounds inhibited invasion by the tumor cells. Gelatin zymography showed that the cells secreted matrix metalloproteinases (MMPs), probably including MMP-2 and MMP-9, which may be involved in tumor cell invasion and metastasis. Theaflavin and theaflavin digallate also inhibited MMPs from the culture medium of these tumor cells, as did (-)-epigallocatechin gallate. These results suggest that theaflavin, theaflavin digallate, and (-)-epigallocatechin gallate inhibit tumor cell invasion by inhibiting type IV collagenases of the LL2-Lu3 cells. PMID- 9339553 TI - Purification and characterization of cysteine proteinase from a baculovirus gene. AB - To analyze the degradation of product proteins at the late stage of virus infection in the baculovirus expression system, a cysteine proteinase was purified from hemolymph of Bombyx mori infected with wild-type B. mori nuclear polyhedorosis virus (BmNPV). The purified cysteine proteinase preparation had two protein bands (major 35-kDa active protein and 28-kDa inactive protein) on SDS PAGE. Based on the N-terminal amino acid sequences of them, it was found that both proteins originated in the cysteine proteinase gene of BmNPV. The purified cysteine proteinase had an optimum pH at 4.0, and also had activities at neutral pHs. When recombinant luciferase was used as a natural substrate, it was degraded rapidly by the cysteine proteinase at the physiological pH of hemolymph. These results suggest that the cysteine proteinase from a BmNPV gene participates in the degradation of foreign protein expressed by the baculovirus system. PMID- 9339554 TI - A novel acid phosphatase from Aspergillus niger KU-8 that specifically hydrolyzes C-6 phosphate groups of phosphoryl oligosaccharides. AB - We had analyzed the detailed structures of the phosphoryl oligosaccharide-1 (PO 1) fraction that was the main component of phosphoryl oligosaccharides (POs) prepared from a potato starch hydrolysate. PO-1 fraction was made up of 3 phosphoryl oligosaccharides and 6-phosphoryl oligosaccharides. Aspergillus niger strain KU-8 produced two types of intracellular acid phosphatase (EC 3.1.3.2, ACPase); ACPase I and II. ACPase II preferentially dephosphorylated 6-phosphoryl oligosaccharides rather than 3-phosphoryl oligosaccharides. The molecular weight of the enzyme was estimated as 66 kDa by SDS-polyacrylamide gel electrophoresis and about 260 kDa by gel filtration, implying the active form to be a tetramer. The optimum pH and temperature of the enzyme were 2.0-2.5 and 60 degrees C, respectively. ACPase II was stable below 50 degrees C for 30 min and pH 2.0-10.0 for 60 min. In spite of the strict specificity toward 6-phosphoryl oligosaccharides in the PO-1 fraction, ACPase II was able to hydrolyze Fru-1,6-di P, ATP, pyrophosphate, and polyphosphate as well as pNPP and Glc-6-P, a broad substrate specificity. PMID- 9339555 TI - Lowering effect of phenolic glycosides on the rise in postprandial glucose in mice. AB - Glycosides were screened for their lowering effect on the postprandial blood glucose rise in vivo. The effect of phlorizin and other phenolic glycosides on the postprandial blood glucose response to glucose ingestion was evaluated in Std ddY mice. When phlorizin was simultaneously added, the peak blood glucose level was significantly decreased by 51% (p < 0.01) compared to vehicles following glucose ingestion by mice, while the blood insulin responses were generally similar. Screening experiments were conducted with different classes of phenolic glycosides added to a glucose solution. Reductions of 40-52% (p < 0.05) were observed in vehicles containing arbutin, 4-hydroxyphenyl-alpha-D-glucopyranoside (hydroquinone-alpha-glucoside) or glycyrrhizin, and of only 15-31% (not significant) in vehicles containing neohesperidin dihydrochalcone, glycyrrhetinic acid monoglucuronide, or 3,4-dimethoxyphenyl-beta-D-glucopyranoside. No lowering effect was observed in vehicles containing salicin. Since glycyrrhizin, arbutin, and hydroquinone-alpha-glucoside blunted to varying degrees the postprandial blood glucose rise following glucose ingestion, they may be useful adjuvants for the treatment of diabetic subjects. PMID- 9339556 TI - Effect of a thyroid hormone treatment on brain protein synthesis in rats. AB - The effect of the thyroid hormone on the rate of brain protein synthesis in rats was studied. Experiments were conducted on three groups of rats given 6-propyl-2 thiouracil (PTU, a thyroid inhibitor) without a triiodothyronine (T3) treatment, those treated with PTU + T3, and those treated with neither PTU nor T3 (control). The fractional rates of protein synthesis in the brain, liver, and kidney of rats given PTU + T3 were significantly greater than those in rats given PTU alone. In the brain and kidney, the RNA activity [g of protein synthesized/(g of RNA.d)] were significantly correlated with the fractional rates of protein synthesis. In the liver and kidney, the RNA concentration (mg of RNA/g of protein) was related to the fractional rate of protein synthesis. These results suggest that the thyroid hormone treatment would be likely to increase the rate of protein synthesis in the brain of rats, and that the RNA activity is, at least partly, related to the fractional rate of brain protein synthesis. PMID- 9339557 TI - Increases in hematopoietic responses caused by beta-glucans in mice. AB - The effects of various (1-->3)-beta-D-glucans on hematopoietic responses of mice were investigated by measuring colony stimulating activity in sera and ascites of the mice administered glucan. We have demonstrated that the hematopoietic response was increased by various structures of (1-->3)-beta-D-glucans, i.e. soluble glucans (linear, branched, single helix, triple helix) and particulate glucans. From the viewpoint of structure and activity relationships, we found several characteristic features: i) hematopoietic response induced by the particulate glucan disappeared faster than that by the soluble glucans, ii) conformation of the glucans, single vs. triple helix, are relatively independent of the response, iii) linear glucan had a weaker response, and iv) there is a strong strain-dependency of the response. These results corresponded well with the fact that branched (1-->3)-beta-D-glucans, but not linear and not particulate, are often used as biological response modifiers for cancer patients. PMID- 9339558 TI - Purification and characterization of a cysteine protease from corms of freesia, Freesia reflacta. AB - A protease (freesia protease B) has been purified to electrophoretic homogeneity from corms of freesia, Freesia reflacta by five steps of chromatography. Its M(r) was estimated to be about 26,000 by SDS-PAGE. The optimum pH of the enzyme was 6.0-7.0 at 30 degrees C using casein as a substrate. The enzyme was strongly inhibited by p-chloromercuribenzoic acid but not by phenylmethanesulphonylfluoride and EDTA. These results indicate that freesia protease B is a cysteine protease. Nine sites of oxidized insulin B-chain were cleaved by freesia protease B in 24 h of hydrolysis. The four cleavage sites among them resembled those of papain. From the digestion of five peptidyl substrates the specificity of freesia protease B was found to be approximately broad, but the preferential cleavage sites were negatively charged residues at P1 positions. Freesia protease B preferred also the large hydrophobic amino acid residues at the P2 position, in a similar manner to papain. The amino terminal sequence of freesia protease B was identical with those of papain in regard to the conservative residues of cysteine protease. PMID- 9339559 TI - Beta ray-induced scission of DNA in tritiated water and protection by a green tea percolate and (-)-epigallocatechin gallate. AB - The beta-ray induced scission of puC18 plasmid DNA from E. coli in tritiated water was examined in the presence or absence of a green tea percolate (TP) and the main constituent, (-)-epigallocatechin gallate (EGCg). An analysis of the ratio of the original closed-circular to the open-circular form of DNA, which was formed by the strand scission of DNA, revealed that TP and EGCg showed a protective effect on DNA scission depending on their concentrations. A new technique, named solid state spin trapping, was applied to examine this scavenging ability toward the hydroxyl (OH) radical generated in tritiated water. The result was kinetically analyzed to reveal that TP and EGCg showed the scavenging effect, suggesting that the protective effect on DNA scission was attributable to the scavenging effect on the OH radical. PMID- 9339560 TI - Purification and characterization of novel whey glycoprotein WGP-88 which binds to a monoclonal antibody to PAS-4 glycoprotein in the bovine milk fat globule membrane. AB - A monoclonal antibody to the PAS-4 glycoprotein (78 kDa) of the bovine milk fat globule membrane (MFGM) specifically recognized PAS-4, and was named KAS4. A component recognized by KAS4 was found in whey protein, this being a glycoprotein of 88 kDa by SDS-PAGE and named WGP-88. WGP-88 was purified and characterized in comparison with PAS-4. WGP-88 had apparent pI values of 6.45 and 6.39, while those of PAS-4 were 7.39 and 7.35. Neuraminidase digestion shifted the pI values of WGP-88 to 6.57 and of PAS-4 to 7.52. WGP-88 was rich in polar amino residues (44.9 mol%), while PAS-4 was abundant in nonpolar amino acid residues (48.7 mol%). WGP-88 contained 17.1% of carbohydrate and PAS-4 had 7.2%. The results of reductive hydrolysis, N-glycanase digestion, and a lectin blot analysis suggested that N- and O-linked sugar chains were contained in both glycoproteins. WGP-88 and PAS-4 had a different N-terminal amino acid sequence. WGP-88 and PAS-4 respectively inhibited competitively the binding of KAS4 to PAS-4 and WGP-88. Our studies revealed WGP-88 recognized by KAS4 mAb to be a novel whey protein and to have different biochemical properties from those of PAS-4. PMID- 9339561 TI - Purification and some properties of lignostilbene-alpha, beta-dioxygenase isozyme IV from Pseudomonas paucimobilis TMY1009. AB - Lignostilbene-alpha, beta-dioxygenase isozyme IV was purified from ultrasonic extracts of Pseudomonas paucimobilis TMY1009 through four steps of column chromatography. The fraction obtained gave a single band on SDS-PAGE and a single peak on reversed-phase HPLC and DEAE-HPLC. The specific activity and Km of purified isozyme IV were 110 mu kat/g and 4.2 microM for 4.4'-dihydroxy-3,3' dimethoxystilbene, 150 mu kat/g and 3.3 microM for 4,2'-dihydroxy-3,3'-dimethoxy 5'-(2"-carboxyvinyl)stilbene. The molecular mass of intact isozyme IV was estimated to be 94 kDa by gel permeation chromatography, and that of its subunits was 52 kDa by SDS-PAGE under denaturing conditions. The N-terminal amino acid sequence of isozyme IV differed slightly from that of other isozymes. Isozyme IV seemed to be composed of two identical subunits, gamma gamma. PMID- 9339562 TI - Orientation of photosynthetic reaction center reconstituted in neutral and charged liposomes. AB - The photosynthetic reaction center from the photosynthetic bacterium Rhodobacter sphaeroides was reconstituted into neutral, positively charged, or negatively charged liposomes. About 70% of photosynthetic reaction centers were reconstituted in the proteoliposomes exposing their H-subunit outside with positively charged lipids while only 30-40% of them were in the same topological orientation with neutral or negatively charged lipids. PMID- 9339563 TI - Transient expression of goat growth hormone gene in poplar (Populus alba L.) protoplasts: a quick method for detection of foreign gene expression in mRNA level. AB - We developed a sensitive, accurate, and fast method to detect foreign gene expression using reverse transcriptase-mediated polymerase chain reaction (RT PCR) in poplar tree (Populus alba L.) protoplasts. Template total RNA was purified by removing the transfected foreign gene vector completely with DNase I treatment before the RT reaction. Expression of cDNA that encodes goat growth hormone was confirmed at the mRNA level 24 h after electroporation mediated DNA transfer. PMID- 9339564 TI - Polyamine content of ordinary foodstuffs and various fermented foods. AB - Soybeans, tea leaves, and mushrooms were conspicuously rich in spermidine, while oranges contained a large amount of putrescine. Among the fermented foods, soy sauces were rich in putrescine and histamine, while Japanese sake contained plenty of agmatine. These polyamines are thought to be produced from amino acids during fermentation with amino acid decarboxylases formed by the micro-organisms. PMID- 9339565 TI - Generation of basidiomycetous hyphal cell-aggregates by addition of the Arg-Gly Asp motif-containing fragment of high-molecular-weight cell-adhesion protein MFBA derived from the basidiomycete Lentinus edodes. AB - The Arg-Gly-Asp (RGD) motif-containing fragment of high-molecular-weight cell adhesion protein MFBA derived from Lentinus edodes caused a significant aggregation of the fragmented hyphal cells of Schizophyllum commune. This fungal cell-aggregation was inhibited by a previous treatment of the cells with the Gly Arg-Gly-Asp-Ser-Pro peptide, but not with the Gly-Arg-Gly-Glu-Ser-Pro peptide, showing that the RGD motif is essential for the cell-aggregation activity. PMID- 9339566 TI - High responsiveness to thyroid hormone of adult rat primary hepatocytes cultured on EHS-gel. AB - The induction of malic enzyme gene expression by triiodothyronine and insulin was severely blunted in rat monolayer hepatocytes cultured on type I collagen compared with that in spherical hepatocytes cultured on a reconstituted basement membrane gel (EHS-gel). Although the mRNA level of thyroid hormone receptor beta (TR beta) gradually decreased in the monolayer hepatocytes during culture, the mRNA level in the hepatocytes on EHS-gel was maintained at around the in vivo level. Our results suggest that the maintenance of TR beta mRNA on EHS-gel is responsible for the high responsiveness to thyroid hormone in a hepatocyte culture. PMID- 9339567 TI - Cloning of a gene encoding a putative xylanase with a cellulose-binding domain from Humicola grisea. AB - We have isolated a genomic clone of a putative xylanase gene (xyn1) from Humicola grisea by using the DNA fragment encoding a cellulose-binding domain of H. grisea cellobiohydrolase 1 as a probe. The translation product of the xyn1 gene predicts a xylanase of 429 amino acids in length, with a cellulose-binding domain in the C terminus. PMID- 9339568 TI - DNA sequence of Bacillus subtilis (natto) NR-1 gamma-glutamyltranspeptidase gene, ggt. AB - The ggt encoding gamma-glutamyltranspeptidase (GGT) from Bacillus subtilis (natto) was cloned and sequenced. The DNA sequence contains a single open reading frame of 1761 bp that might be translated to a protein of 587 amino acid residues, and indicates that B. subtilis (natto) GGT is synthesized as prepro-GGT and processed later into large and small subunits. The putative catabolite responsive element (CRE) was located upstream of the ggt coding region. PMID- 9339569 TI - A ribosome-inactivating protein from Amaranthus viridis. AB - An antiviral protein purified from the leaves of Amaranthus viridis was named amaranthin. The in vivo antiviral activity of amaranthin was confirmed in tobacco mosaic virus (TMV) infection test on Nicotiana glutinosa leaves. The molecular mass of the amaranthin was estimated about 30 kDa by SDS-PAGE and the pI was measured as 9.8 by isoelectric focusing (IEF) analysis. Cytotoxicity of the amaranthin using in vitro translation inhibition assay was similar to that of pokeweed antiviral protein (PAP) with IC50 of 25 pM. Depurination activity (N glycosidase activity) against animal rRNA was also confirmed. PMID- 9339585 TI - Stroke--changing the focus. PMID- 9339586 TI - Stroke. A looming epidemic? AB - A recent deceleration in the long established downwards trend in mortality from stroke combined with rapid ageing of the Australian population suggests that we may be facing a large and rapid increase in the caseload of stroke unless efficacious ways to prevent cerebrovascular disease are applied more systematically. This paper reviews the current situation, drawing on evidence from the Perth Community Stroke Study (PCSS). PMID- 9339587 TI - Haemorrhagic stroke. Intracerebral and subarachnoid haemorrhage. AB - A proportion of strokes are due to intracranial haemorrhage. Their characteristics and treatment are different from ischaemic stroke-surgery is often necessary and thrombolytic or anticoagulant treatment contraindicated. Most intracerebral haematomas (ICH) are hypertensive in origin. Small haematomas are treated with blood pressure control and rehabilitation. Larger ones often need surgery. Treatment must be tailored to the patient's circumstances, taking into account age, general and neurological health, location of haematoma etc. Subarachnoid haemorrhage (SAH) affects younger people and is classically due to aneurysm rupture, requiring urgent neurosurgical referral. Another cause is arteriovenous malformations (AVM), also treated surgically. A major problem is failure to diagnose less severe bleeds, which often precede major haemorrhages. PMID- 9339588 TI - Current management of acute stroke. AB - Acute stroke is an emergency clinical situation that is relatively frequently encountered by general practitioners. Recent studies in the management of cerebral ischaemia have determined that the time to definitive treatment is an important factor. Thus, the actions of the medical professional who has initial contact with the patient may significantly affect the outcome of the patient with stroke. PMID- 9339589 TI - Rehabilitation. Does it make a difference? AB - Stroke is a major cause of disability. Comprehensive multidisciplinary assessment and treatment programs have been proven to improve survival and functional outcome compared with less organised systems of stroke care. Patients who undergo such rehabilitative programs are more likely to be alive and living at home 1 year later. PMID- 9339590 TI - Learning relevant procedural skills. Are supervisors providing opportunities? AB - OBJECTIVE: The aim of this study is to assess general practice registrars' (GPRs) learning needs in relation to their procedural skills development and compare these with the learning opportunities offered by their general practice supervisors (GPSs). METHOD: A 43 item questionnaire was designed to assess the procedural skills development needs of GPRs. The questionnaire was administered to GPSs and GPRs. RESULTS: GPRs can expect to consolidate their skills in the areas of joint injections. ENT procedures and some small surgical procedures. They are unlikely to gain experience in the areas of proctology, the use of the microscope as a diagnostic tool, cardiac stress testing, hearing assessment and some less frequently needed procedures. Respondents identified spinal mobilisation manipulation, flexible sigmoidoscopy, colposcopy and fine needle aspiration biopsy as procedures they would like to see performed in general practice. CONCLUSION: GPRs are keen to enhance their procedural skills in a wide range of areas. They cannot necessarily expect that their general practice attachment will provide the opportunities to develop all skills desired. The Royal Australian College of General Practitioners (RACGP) Training Program needs to provide registrars with a list of core skills that are expected of every general practitioner (GP). To ensure this objective is achieved, innovative learning opportunities for GPSs and GPRs in the domain of psychomotor competencies need to be developed. PMID- 9339591 TI - Through the looking glass. Computers and general practice. PMID- 9339592 TI - Primary skin infections. PMID- 9339593 TI - Reflections on TIA. PMID- 9339594 TI - Osteoporosis. Guidelines for general practitioners. Osteoporosis Australia. AB - Osteoporosis and its associated problems are major health concerns in Australia, and are commonly encountered in general practice. As the population ages, the incidence of osteoporosis will continue to increase. Hence, it is important for general practitioners and healthcare professionals to remain up to date with the strategies involved in the prevention and treatment of osteoporosis. Over the last few years, there have been significant advances in our understanding and treatment of osteoporosis. It is now clear that osteoporosis is a problem for men as well as for women. Accurate and reliable measurements to diagnose osteoporosis, and effective medications to treat it, are now available. These guidelines have been prepared to help general practitioners identify people at risk of developing osteoporosis, and to inform general practitioners on the current diagnosis and treatment strategies of the condition. We commend these guidelines to you as a reference source for the treatment of people with osteoporosis. PMID- 9339595 TI - Deep vein thrombosis and pulmonary thromboembolism. PMID- 9339596 TI - Sports medicine. Persistent shoulder pain. PMID- 9339597 TI - A cautionary tale. AB - Type 2 diabetes is increasing in prevalence and is predominantly managed in general practice. This series contains case histories raising some of the metabolic problems which may be encountered in the management of these patients and indicates some of the many issues, other than glycaemic control, that need to be considered. PMID- 9339598 TI - A case of persistent cough. PMID- 9339599 TI - Bones and silence. PMID- 9339600 TI - Practice compared--payment systems. PMID- 9339601 TI - Head lice. PMID- 9339602 TI - Australia's notifiable diseases status, 1996. Annual report of the National Notifiable Diseases Surveillance System. AB - In 1996 there were 65,024 notifications to the National Notifiable Diseases Surveillance System. The record high number of Ross River virus infection notifications was of particular note. The highest rates were recorded in Western Australia, where an outbreak was documented in the South West, and in Queensland. Most cases occurred in the late summer and early autumn months. The number of measles cases has continued to fall markedly following the outbreak in 1993 and 1994. Rubella notifications also fell in 1996. The number of cases of pertussis remained at a similar level to that recorded in recent years, the highest notification rate being recorded for children under the age of one year. A peak in late 1996 marked a resurgence in the pertussis epidemic which has continued into 1997. Notifications of Haemophilus influenzae type b continued to decline reaching a record low rate of 0.3 notifications per 100,000 population. For the enteric diseases, the number of cases of campylobacteriosis rose, with an annual adjusted notification rate of 100.4 per 100,000 population; more notifications were received for this disease than for any other in 1996. The number of hepatitis A cases also rose relative to 1995. This is a reversal of the trend observed in recent years when the notification rate fell. The number of cases of salmonellosis and shigellosis remained stable. Notifications for chlamydial infection and gonococcal infection rose relative to 1995, whilst those for syphilis fell. PMID- 9339603 TI - Possible foodborne outbreak. PMID- 9339604 TI - Communicable diseases surveillance. PMID- 9339605 TI - Bronchial asthma and sleep disturbances. PMID- 9339606 TI - Legionella as a lower respiratory pathogen in north India. AB - One hundred patients of lower respiratory tract infection (LRTI) were prospectively studied over 2 years to find out if Legionella is a causative agent in these patients. In addition, 50 environmental samples and 50 age and sex matched controls were studied. Culture and direct fluorescent antibody testing (DFA) of respiratory tract secretions, and serodiagnosis by indirect immunofluorescence (IIF) and ELISA, were employed to detect Legionella. Respiratory tract secretions from all patients were negative for Legionella on culture and DFA. Low antibody titters to Legionella were observed in 21 patients and these could be attributed to cross reaction with other gram-negative bacteria. All environmental samples and controls tested negative for Legionella. Legionella does not seem to be an important lower respiratory tract pathogen in this part of the country and empirical addition of erythromycin to treatment regimens for pneumonia is not warranted in our setting. PMID- 9339607 TI - Peak expiratory flow rate in school-going children. AB - Peak expiratory flow rate (PEFR) was measured with mini Wright's peak flow meter in 783 children (aged 6-17 years) from a school in urban Delhi and 523 children (aged 6-15 years) from another school in Nellore, Andhra Pradesh. In all the children, age in completed years, sex, height, weight, chest circumference at full inspiration and maximum chest expansion were recorded. Age, sex, height and weight were independent predictors of PEFR in children from Nellore. Age, sex and height, were independent predictors of PEFR in boys from Delhi while height alone was an independent predictor of PEFR in Delhi girls. Common prediction equations for predicting PEFR in boys and girls have been developed for both regions based on age and height. For the same height and age, boys had higher PEFR than girls. In the females, the PEFR seemed to have a plateaux effect after the age of 14 years; such an effect was, however, not seen in the boys in the age range studied. The PEFRs of children from both parts of the country were similar, and were lower than those reported for American white children. PMID- 9339608 TI - Ventilatory lung function tests in school children of 6-13 years. AB - Spirometry was performed in 186 healthy school children, 6-13 years of age, selected from urban population of Rohtak city (India), to derive the regression equation for prediction of normal value of ventilatory lung function in this age group. Values of lung functions in the present study are well comparable to other North Indian studies and Western reports, but higher than the South Indian children. Lung functions have shown better linear correlation with age and height as compared to weight. Comparison of ventilatory function tests in boys and girls showed statistically higher FVC, FEV1, PEFR, FEF25% and MVV in boys, while FEF50% FEF75% and FEF75-85% were statistically higher in girls in different age groups. No specific pattern of variation in lung function tests was observed with sex. Regression equations for different ventilatory functions have been derived for either sex with the best possible combination of physical parameters. PMID- 9339609 TI - Use of LPC antagonist, choline, in the management of bronchial asthma. AB - The present study was conducted to compare the effects of disodium cromoglycate (DSCG) and choline, a lipotropic factor, in management of bronchial asthma. Two groups taking DSCG and choline were compared between themselves and with a control group (on bronchodilators only) in a three month trial. A significant improvement in average symptoms scores was observed in all groups but was clinically significant only in the groups taking choline. Percent asymptomatic days increased in all the groups, but was not significant. The additional drug requirement decreased significantly only in the group taking choline. All groups showed an improvement in SGaw at FRC and percent fall in specific airways conductance (SGaw) at RV, but non-significant. A significant fall in bronchial hyperreactivity (BHR) was observed in the group taking choline, which was not consistent, whereas a marginal increase in BHR in the control and DSCG groups was observed. It is concluded that choline, an anti-inflammatory agent which acts by lowering lipophosphatidyl choline (LPC), plays an active role in subjective as well as functional improvement in bronchial asthma. However, a dose related response is yet to be established. PMID- 9339610 TI - Benign giant cell tumour with calcified lung metastasis. PMID- 9339611 TI - Diagnostic dilemma: tuberculosis? or, sarcoidosis? PMID- 9339612 TI - Wegener's granulomatosis: a case with laryngeal involvement. AB - A patient of Wegener's granulomatosis (WG) with laryngeal involvement and respiratory difficulty has been described. Direct laryngoscopic biopsy led to the diagnosis of Wegener's granulomatosis. The patient underwent a tracheostomy and was subsequently treated with cyclophosphamide and corticosteroids to which he responded. PMID- 9339613 TI - Carcinoid tumour: laser therapy. AB - A 31-year-old female, pregnant (I trimester), presented with symptoms and signs of bronchial asthma. Chest roentgenogram and computerized tomography of the chest revealed right mid and lower zone opacity (collapse). Bronchoscopic examination revealed intrabronchial growth in the right middle and lower lobe bronchus. Biopsy and histopathological examination confirmed carcinoid tumour. These symptoms and signs disappeared after laser therapy. The lesion, however, recurred two years later. PMID- 9339614 TI - Malaria: persistent killer: continuing renal complications. PMID- 9339615 TI - Intra-operative suggestions reduce incidence of post hysterectomy emesis. AB - The influence of therapeutic intraoperative auditory suggestions on the incidence and severity of emetic episodes was investigated in 50 adults ASA I and II patients undergoing elective abdominal hysterectomy. The patients were randomly divided into two groups, each consisting of 25 patients. In group I, a blank tape was played and in group II, positive suggestion was played via headphones throughout the anaesthetic period. It was observed that there was statistically significant difference (P < 0.05) between the incidence of vomiting in group I (60%) and group II (36%). The number of vomiting episodes per patient in group I was 3.1 +/- 1.2 as compared to 1.7 +/- 0.6 in group II. This difference was statistically significant. The patients requiring rescue antiemetic was significantly higher (P < 0.05) in group I (66.6%) as compared to group II (22.2%). It is concluded that positive therapeutic suggestion may be considered as an alternative to antiemetic therapy. PMID- 9339616 TI - Prevalence and severity of viral hepatitis in Pakistani pregnant women: a five year hospital based study. AB - A hospital based observational study was carried out on pregnant women presenting with either acute hepatitis or fulminant hepatic failure (FHF), during the past years. Of 53 patients, 20 (38%) developed FHF.Non-A, Non-B was the commonest cause (62%) followed by hepatitis B in 17% and hepatitis A in 4% cases. Eight women expired (case fatality rate 15%) with a high maternal mortality (62%) caused by NANB hepatitis. Perinatal mortality was 30%. Poor prognostic factors identified were lack of antenatal care, severity of jaundice, history of somnolence, gastrointestinal bleeding and a high grade of encephalopathy. PMID- 9339617 TI - Does training affect quality of diarrhoea case management. AB - Improvement in diarrhoea case management through training of health care providers in the government and the private sector was the key element of diarrhoea policy in Pakistan in 1989. Numerous training sessions were organized by the Child Survival project. The aim of this project was to look at the effect of training on quality of diarrhoea case management at the oral rehydration therapy (ORT) corners and diarrhoea treatment units (DTUs) of Sindh. A systematic random sample of 62 ORT corners and DTUs in Sindh was assessed using the WHO drafted Health Facility Survey manual. It was observed that the trained health providers were better in taking history for blood in stools (P < 0.004) and other illnesses (p < 0.000). For assessment of dehydration, no significant difference (p < 0.933), was found between trained and untrained health providers. Trained were better than untrained (p < 0.035) only in treatment "Plan A" and correct rehydration (p < 0.004) of child at facility. Training did not influence advising mothers for home case management. Thus diagnosis was good and training did improve the quality of assessment of child but treatment (inclusive of advice giving) was not significantly affected by training, except for a child with no dehydration. It is recommended that on the job training should emphasize on skills for management of diarrhoea. Further studies are needed to identify why inspite of training. Health providers do not offer better treatment than the untrained ones. PMID- 9339618 TI - Supervision for diarrhoea case management at the oral rehydration therapy corners and diarrhoea training units in Sindh. AB - The role of supervisors in diarrhoea case management at the Oral Rehydration Therapy (ORT) corners of Sindh was assessed by a random sample of 62 ORT corners in rural and urban areas of Sindh. The supervisors at each facility included, Medical Superintendents, District Health Officers, Senior Medical Officers, Civil Surgeons and Medical Officers. In 65% cases the supervisors were working at the same facility and in 35% the supervisors were based elsewhere. In 33.7% cases, there had been no meeting with the supervisor in the past 3 months which incidently was the diarrhoea season. In 6.5% cases there had been only 1 meeting, in 8.5% there had been 2-3 meetings and in the remaining 41.3% cases there had been more than 3 meetings with the supervisor. In majority of the cases the supervisors did not do anything related to improvement of diarrhoea case management. Supervisors had given some importance to case management in the urban areas and the Diarrhoea Training Units (DTU). The supervision at ORT facilities was found to be at a rudimentary level and it is suspected that this might be adversely affecting the quality of diarrhoea case management. PMID- 9339619 TI - Ambulatory arthroscopic knee surgery results of partial meniscectomy. AB - In an open prospective study the results of arthroscopic partial meniscectomy were reviewed in 188 knees. Group I comprised of 139 (65%) knees with pure meniscus lesions without any ligament laxity, the remaining 49 (35%) knees in group II had anterior cruciate ligament (ACL) deficiency in addition to meniscus lesions. In group I, 93% had excellent to good and 7% had fair results by criteria of Tapper and Hoover. In group II, 75% had excellent to good and 25% had fair results. Arthroscopic knee surgery was a good method of identifying patients in group II who required anterior cruciate reconstruction. One hundred (53%) sedentary workers were able to return to work at a mean time of 3 weeks. Patients classified as heavy labour comprised of a group of 40 (21%) were able to return to their occupation in the mean time of 5 weeks. School and college students numbered 28 (14%) were able to go to their institutions in the mean time of 10 days. Twenty (9%) patients were professional or semi-professional athletes who were permitted sports training in 3 weeks and sports participation in 6 weeks on the average. There were no serious complications including wound infection, deep venous thrombosis (DVT) or loss of knee motion. The arthroscopic technique is reliable, cost effective and has high patient acceptance because of low morbidity and rapid return of good function to the knee joint. PMID- 9339620 TI - Epidemiology of cerebral malaria and its mortality. AB - Over the past 5 years, 1620 comatosed patients of both sexes aged 1-75 years were screened for cerebral malaria. Of these, 505 (31.2%) were positive for Plasmodium falciparum. During this period frequency of malaria increased from 22.1% in 1991 to 44.4% in 1995. Sixty-four percent cases of cerebral malaria were seen in children and thirty-six percent in adults. Mortality was also higher (41%) in children than in adults (25%). As cerebral malaria is particularly prevalent in Pakistan and is a major community problem, accurate and easier methods of its diagnosis are needed at primary health care level, in all febrile comatose patients, without focal neurological findings. PMID- 9339621 TI - Immunoproliferative small intestinal disease and primary small intestinal lymphoma: review of literature. PMID- 9339622 TI - Sarcomatoid variant of renal cell carcinoma. PMID- 9339623 TI - [An epidemiological investigation for gram-positive coccus, especially MRSA, in Kinki area]. AB - An epidemiological investigation for Methicillin-resistant Staphylococcus aureus (MRSA) was performed at 21 medical institutes in Kinki area by the questionnaire from Kinki Infection Working Group in 1995. The most frequent specimen for MRSA was pus (25.3%) from out-patients and sputum (34.4%) from in-patients, respectively. As compared to the investigation of the MRSA Forum in 1992 and 1993, MRSA increased over 20%, and furthermore 57.8% was MRSA out of S. aureus under this study. It demonstrated that drug-resistance was accelerated for a few years. Polymicrobial infection, especially with more than 3 pathogens, increased significantly compared with the results of the MRSA Forum (p < 0.01). It was suggested that compromized host increased and therapy turned to be more difficult. Simultaneous pathogens detected with MRSA were Candida species, Pseudomonas aeruginosa and Enterococcus faecalis. In terms of chemosusceptibility, VCM (99.9%), ST (99.4%) and ABK (99.3%) were still determined to be highly sensitivity against MRSA. PMID- 9339624 TI - [An epidemiological investigation for gram-positive coccus, especially PRSP, in Kinki area]. AB - An epidemiological investigation for penicillin-resistant Streptococcus pneumonia (PRSP) was performed at 18 medical institutes in Kinki area by the questionnaire from Kinki Infection Working Group 1995. This investigation was the first report that was performed for a long term (one year) and a large area. The most frequent specimen was sputum from out-patients (50.3%) and inpatients (48.8%), and especially from spinal fluid of 3 cases were detected. Polymicrobial infection with more than 3 pathogens was 15.7%, and it was more frequent than MRSA previously investigated. Simultaneous pathogens detected with PRSP were Candida species, Haemophilus influenzae and Staphylococcus aureus. In terms of chemosusceptibility, VCM (100%), FMOX (97.9%), IPM/CS (85.9%), CEZ (93.4%) and CDTR-PI were determined to be high by sensitive. However, the sensitivity of CCL, which was one of the most common antibiotics, was only 37.7%. PMID- 9339625 TI - [Analysis of genome types of adenovirus type 7 isolated in Fukuoka Prefecture in 1996]. AB - Adenovirus type 7 (Ad7) was rarely isolated in Japan till 1994, but during April 1995 to August 1996, isolations of Ad7 were reported 230 cases. We isolated Ad7 in January and July 1996 in Fukuoka prefecture. We analyzed its genome type by using 14 restriction endonuclease and studied seroepidemiology of Ad7 infection in Fukuoka prefecture. Isolated Ad7 strains were identical by 14 restriction endonuclease. Between new Ad7 isolates and prototype (Gomen; Ad7p), 4 restriction endonuclease patterns were identical but 10 restriction endonuclease patterns were different. From the result of restriction endonuclease pattern analysis, genome type of Ad7 isolated in Fukuoka may be the same to Ad7c reported by Noda et al. (1996). The alterations in the cleavage sites of 10 restriction endonucleases between new Ad7 isolates and Ad7p were revealed at least 12 sites. Ad7 antibody positive rates in serum specimens collected in Fukuoka Prefecture were 3.6% in 1994 and 9.7% in 1996. PMID- 9339626 TI - [Relation between candidiasis and nutrition of patients and MRSA infection]. AB - We evaluated the relation with nutritional state and methicillin-resistant Staphylococcus aureus (MRSA) infection of 20 patients who isolated Candida of 31 patients who were suspected as candidiasis. All of the patients who were hospitalized in internal medicine wards in our hospital between February 1993 and January 1994. All patients with hematological disease were excluded. MRSA is now gotten much attention as pathogen of hospital infection, but each MRSA is detected not only alone but also concomitant with other bacterium. In our hospital MRSA was detected alone 36.8% (39/106). Candida 9.4% (10/106) was frequent as mixed infectious pathogen concomitant with MRSA following Pseudomonas aeruginosa 19.8% (21/106) and Enterococcus faecalis 12.3% (13/106). Intension of host-immunity is a important factor as prevention of both infection. We evaluated the nutritional state of patients with 7 cases of infection group and 13 cases of colonization group. There was no significance between two groups as for serum total protein (mean) were 5.97 g/dl in infection group and 5.96% g/dl in colonization group. Furthermore there were significance (p < 0.01) for mean serum albumin (mean) were 3.11 g/dl v.s. 3.27 g/dl. But both serum total protein and serum albumin of these evaluated patients who isolated Candida were less than normal limit. Therefore we considered nutritional disturbance was important addition to over-use of antibiotics and severe underlying diseases as risk factor of candidiasis, and revealed the importance of improvement of nutritional state at the prevention and therapy. PMID- 9339627 TI - Differential diagnosis of Trypanosoma cruzi and T. rangeli infection by PCR of cysteine proteinase genes. AB - Trypanosoma cruzi is the causative parasite of Chagas' disease, while the closely related T. rangeli is non-pathogenic in humans. Restriction fragment length polymorphisms (RFLPs) of a portion of the cysteine proteinase gene were used to distinguish T. cruzi from T. rangeli. This procedure was very sensitive, with a single cell of either species being sufficient for the PCR. The sensitivity and clear ability to distinguish between T. cruzi and T. rangeli suggest that this procedure may be easily applied in the field. PMID- 9339628 TI - [Effects of various steroidal and non-steroidal anti-inflammatory drugs on in vitro IL-10 production of murine peritoneal macrophages infected with Mycobacterium avium complex]. AB - In murine infections due to Mycobacterium avium complex (MAC), bacterial regrowth of the pathogens is frequently encountered in the relatively late phase of infection even in mice receiving daily treatments with antimicrobial agents including rifamycins and macrolides. In this case, the bacterial regrowth is usually accompanied by concomitant increase in the tissue levels of IL-10 and transforming growth factor-beta (TGF-beta), well known immunosuppressive cytokines. In this context, it is of interest to note recent findings that steroidal anti-inflammatory drugs up-regulate TGF-beta production from T lymphocytes, thereby suggesting participation of immunosuppressive cytokines in the expression of their anti-inflammatory activity. In this study, we examined the effects of various anti-inflammatory drugs including glucocorticoids and non steroidal anti-inflammatory drugs (NSAID) on in-vitro IL-10 production of murine peritoneal macrophages (M phi s) infected with MAC organisms. When the IL-10 production by MAC-infected M phi s was measured in terms of protein and mRNA expression of the cytokine using ELISA and RT-PCR assays, the following results were obtained. First, the IL-10 production into M phi culture fluids was temporarily increased around days 1 to 3, thereafter gradually declined, and returned to normal by day 14. Secondly, IL-10 mRNA expression of M phi s was rapidly induced after MAC-infection and the maximum level of the mRNA expression was achieved at 2 hr. Thereafter, IL-10 mRNA expression ceased rapidly and returned to normal by 24 hr. Thirdly, the majority of test anti-inflammatory drugs, not only glucocorticoids but also NSAID, were found to up-regulate the IL 10 production of MAC-infected M phi s, suggesting some possible roles of IL-10 in the expression of the anti-inflammatory activities of these agents. PMID- 9339629 TI - [Simple method for typing human immunodeficiency virus type I (B, E) by PCR and chronological change of distribution of subtype (B to E) in Japan]. AB - We analyzed the clade distribution of B and E in HIV-1 isolates in Japan by a nested PCR method using 5'-CCCACAAGATTTAAATATG-3' of the gag gene as clade B primer and 5'-CCCACAAGATTTAAACTCC-3' of the gag gene as clade E primer. Seventy two anti-HIV-1 confirmatory positive serum samples were collected during a period of 1991-1996 in two hospitals in Yokohama City. Peripheral blood mononuclear cells were obtained from the buffy coat of these samples and extracted DNA were used for nested PCR. The 72 cases comprised of 11 Japanese hemophiliacs, 14 Japanese male homosexuals, 19 Japanese male heterosexuals, 5 Japanese female heterosexuals and 23 Thai female heterosexuals. Of these 36 were clade B and 35 were clade E and one case showed positive PCR results for both B and E primers. Almost all male Japanese hemophiliacs and homosexuals in our sample have clade B, while the female Thai heterosexuals have clade E, irrespective of the year of isolation. As for Japanese male heterosexuals, through 1993, clade B was predominant but since 1994, the predominate clade switched to clade E. Although the number of Japanese female heterosexuals in our sample is small, clade B was isolated in 2 cases even after 1994. PMID- 9339631 TI - [Studies of Vibrio cholerae O140 (serogroup Hakata) isolated from river water]. AB - Strains of Vibrio cholerae O140, which were isolated from river (Hikichi River) water in Kanagawa, Japan, were studied for their biochemical characteristics and toxin genes, and analyzed by an amplified polymorphic DNA method in comparison with both clinical and environmental isolates of V. cholerae O140. Biochemical characteristics of 4 river isolates studied were found to be unanimous by conventional tests and a commercially available identification kit (Api 50 CH). The strains were also found to react positively with both cholera Inaba factor (C) and V. cholerae O140 specific factor (F). A total of 34 V. cholerae O140 strains including the 4 stains, a reference strains (487-95), 25 strains isolated from samples of river or sea water in Japan, 4 strains isolated from a patient and imported foods, was investigated by the PCR method for presence of ctx, zot, hlyA, and NAG-ST genes. It was found that all strains carried the hlyA gene but none of them carried either ctx, zot, or NAG-ST genes. V. cholerae O140 shared a common antigenic structure with pathogenic V. cholerae strains but its pathogenicity seems to be comparable to that of V. cholerae non-O1 non-O139. Subsequent RAPD analysis indicated that the strains can be divided into 9 groups. Four strains from the Hikichi River showed the same RAPD pattern. The results suggest that the analysis can be a useful epidemiological tool for V. cholerae O140, and that V. cholerae O140 persists in the Hikichi River throughout years. PMID- 9339630 TI - [A study for serodiagnosis of verotoxin-producing Escherichia coli (enterohemorrhagic E. coli) O157 by the bacterial agglutination technique]. AB - We evaluated the usefulness of bacterial agglutination antibodies for serodiagnosis of verotoxin-producing Escherichia coli (enterohemorrhagic E. coli) O157 infections. We examined 50 serum samples from 50 control children (whiout diarrhea 31, with diarrhea 19), 24 samples from 8 diarrhea cases due to O157:H7, 37 samples from 14 cases of hemolytic uremic syndrome (HUS) for antibodies to heat-killed E. coli E32511 (O157:H.-) strain using the bacterial agglutination technique. Of the control sera all but one (x80) showed 20 > or = in the antibody. All the diarrhea patients due to O157:H7 showed a significant rise (x160-x5120) of the titers in the sera at 5-7 days on illness, after that the titers fell rapidly. Significant antibody rise (x160-x5120) was detected in twelve out of 14 HUS patients at the early stage of the illness which fell in the convalescent phase. The assay appeared to be a useful serodiagnostic technique because of its easiness and simplicity as well as because of its high sensitivity and specificity. PMID- 9339632 TI - [A case of hemolytic uremic syndrome documented co-infection of vertoxin producing Escherichia coli O157 and other pathogens]. AB - We reported a case of a 3-year-old girl with hemolytic uremic syndrome (HUS), which showed hemolytic anemia (Hemoglobin 8.2 g/dl, lactate dehydrogenase 1277 IU/l and total bilirubin 0.6 mg/dl), small purpura on the skin (platelet 7.3 x 10(4)/microliter) and slightly decreased output of urine (creatinine 0.4 mg/dl and blood urea nitrogen 27.2 mg/dl). Verotoxin producing Escherichia coli (E. coli) O157 was not isolated, but Salmonella agona and E. coli O125, which is one of the enteropathogenic E. coli, were detected from her stool culture. However, the IgM antibody against verotoxin producing E. coli (VTEC) O157 lipopolysaccharide was detected in both serum of the acute and convalescent phase by immunoblot assay. In addition verotoxin DNA was demonstrated in the stool by PCR method. Therefore, we think this HUS might be due to VTEC O157, which must have been co-infected with Salmonella agona and E. coli O125. There have been four cases including the present case of co-infection with VTEC O157 so far, and the other three cases were of the Salmonella species. Although the reason of co infection was unknown, we may infer that food might be contaminated with some pathogens including Salmonella species or that these patients might be already infected with Salmonella species prior to VTEC infection. Even when some other pathogens were detected by a stool culture from a patient with HUS, we should pay attention to demonstrate associated of VTEC and HUS by the specific antibodies and PCR for verotoxin DNA. PMID- 9339633 TI - [A case of multiple muscular abscesses of the lower limbs by Staphylococcus aureus after chemotherapy for lung cancer]. AB - A 67-year-old male was admitted to our hospital because of lung cancer and interstitial pneumonia. Cisplatin, vindesie and mitomycin C were administered for treatment of lung cancer. The leucocyte-counts declined to 1700/microliter on the eighth day after the chemotherapy. Though granulocyte colony-stimulating factor was administered, pain in the right thigh and high grade fever developed. Because Staphylococcus aureus was isolated from the blood specimen, piperacillin was administered. But the high grade fever continued and the pain was expanded to the right hip, left hip, thigh and leg. Because a computed tomograph of the lower limbs showed low density areas in bilateral gluteus maximus muscle right adductor magnus muscle, left biceps femoris muscle and left soleus muscle and the culture of an aspirate from abscess of right leg detected S. aureus, multiple muscular abscesses of the lower limbs was confirmed. We changed the antibiotics from PIPC to imipenem/cilastatin and minocycline on nineteenth day after the chemotherapy. His symptoms improved after the change of antibacterial agents. But he died of acute exacerbation of interstitial pneumonia, after about two months of the chemotherapy. Muscular abscesses of the limbs are very rare in Japan. Only four cases with muscular abscess of the limbs were reported in Japan, since 1988. This case suggests that a muscular abscess must be considered in the differential diagnosis of fever in patients with neutropenia. PMID- 9339634 TI - [A case of an elderly patient with dementia and gait disturbance associated with influenza]. AB - We report a case of progressive dementia and prolonged gait disturbance correlated with influenza A/H3N2 infection in 91-year-old female patient, admitted because of in ability to take care of herself due to aging and cerebral infarction. At admission, conversation and comprehension were not significantly impaired, and she was able to walk by herself. Flu symptoms such as high grade fever, chills, arthralgia, and cough appeared after a short stay at home. Influenza A/ H3N2 was confirmed serologically. Delirium occurred on the sixth day after influenza onset, persisted for three weeks, followed by recovery. Dementia symptoms such as memory defects and disorientation continued and did not improve. Due to this febrile episode, she was unable to walk unassisted. The results of computed tomography performed before and after the influenza episode were unremarkable for additional cellebro-vascular events during the observed period. Influenza infection may be an important risk factor for reducing the quality of life in the elderly. In geriatric cases, influenza should not be treated as a mere transient illness, but rather one which has important consequences for the elderly population, including the possibility of life threatening complications. PMID- 9339635 TI - [A case of primary HIV-1 infection]. AB - A previously healthy 28 year old Japanese man came to us with a genital ulcer which appeared 13 days before admission to our hospital. He had subsequently fever (40 degrees C), arthralgia, a sore throat and oral aphtha 6 days before admission. He had a history of sexual contact with a female commercial sex worker one week before his illness. On the day of admission, he had shallow ulcers on the lip, tongue and penis. Initial laboratory test included leukopenia and thrombocytopenia. His fever abated 3 days after admission. His condition and bicytopenia recovered completely after 12 days of admission. Although, his serum HIV-1 antibody was negative when he was admitted, 3 months later the antibody was seroconverted. And p24 antigen and HIV-1 RNA of stocked serum were positive. Diagnosis of primary HIV-1 infection was made. Recently, HIV-1 infection has been increasing in Japan. Consideration of this disease in differential diagnosis of acute febrile illness is necessary. PMID- 9339636 TI - Interpersonal psychotherapy: current status. AB - Interpersonal psychotherapy (IPT), a time-limited treatment for major depression, was developed, defined in a manual, and tested in randomized clinical trials by the late Gerald L Klerman, MD, and collaborators. It has subsequently been modified for different age groups (adolescents-elderly), types of mood disorders (dysthymia, bipolar disorder, antepartum and postpartum patients), and non-mood disorders (bulimia, drug abuse, borderline personality disorder, social phobia, somatization, medically ill patients). It has been used as a long-term treatment, in a group format, over the telephone, and as a patient guide. It has been translated into Italian, German and recently Japanese. Having begun as a research intervention, IPT is only recently being disseminated among clinicians or in residency training programs. The publication of efficacy data, the appearance of two practice guidelines in the United States that include IPT among treatments for depression, the interest in defined treatments for managed care and the endorsement of Consumers Guide have led to increasing requests for information and training. This paper briefly describes the concepts and techniques of IPT and the current status of adaptation. In summary, evidence from controlled clinical trials suggests that IPT is a reasonable alternative or adjunct to medication as an acute, continuation, and/or maintenance treatment for patients with major or mild depression, patients who are human immunodeficiency virus (HIV) positive, or who have bulimia. It is a promising treatment for depressed adolescents and for geriatric patients, for patients with dysthymia and as treatment for depressed couples marital disputes. A final conclusion awaits the completion of clinical trials underway before substantial claims can be made. IPT is not effective, as compared to a standard drug program, for opiate and cocaine addicted patients. PMID- 9339637 TI - Ligand recognition by beta 3 integrins. AB - Integrins are alpha/beta heterodimeric adhesion receptors, alpha IIb beta 3 (GPIIb-IIIa) and alpha v beta 3 (the vitronectin receptor) share the same beta subunit, beta 3, but have distinct alpha subunits. These sister integrins not only recognize many of the same ligands but also have certain unique ligands. Based upon current information in the literature, we propose that four classes of beta 3 ligands can be distinguished. Since the beta 3 integrins have multiple functions in vivo and are targets for therapy, this classification system may be useful in the design and characterization of therapeutic agents. PMID- 9339639 TI - Rapid changes in pial arterial diameter and cerebral blood flow caused by ipsilateral carotid artery occlusion in rats. AB - We investigated rapid changes in pial arterial diameter and in cerebral blood flow (CBF) caused by transient ipsilateral common carotid artery occlusion (CCA O) in anesthetized rats in order to elucidate how the cerebral circulation reacts to acute stem artery occlusion. In separate groups of rats, pial arterial diameter was recorded through a closed cranial window and CBF was recorded by laser-Doppler flowmetry. CCA-O was performed for 5 minutes under normotension and normocapnia (control) and under graded hypotension, hypercapnia and hypocapnia. In the control condition, pial arterial diameter increased rapidly, triggered by CCA-O. It took 12 +/- 3 s to reach the maximum of 204 +/- 42% of the value before CCA-O, and 60 +/- 24 s to become stable at 131 +/- 11%. CBF decreased rapidly to 66 +/- 11%, then increased reactively to 135 +/- 9%, and again decreased to 91 +/ 3%. The reactive increase in CBF caused by CCA-O decreased in parallel with the degree of hypotension, and also became barely detectable under hypercapnia. Our data suggest that active vascular dilation in the territory of the occluded artery is important for inducing collateral circulation. PMID- 9339638 TI - Prevention of aminoglycoside-induced hearing loss. AB - This review discusses the current problem of ototoxicity associated with the worldwide use of aminoglycoside antibiotics. Pathology and pathophysiology of cochlear and vestibular damage have long been recognized as a preferential destruction of hair cells beginning in the cochlea with the outer hair cells of the lower turns. This is accompanied by high frequency hearing loss progressing to lower frequencies during and even after treatment with these drugs. A novel hypothesis of the underlying biochemical mechanism is based on the formation of free radicals by an aminoglycoside-iron complex. A protective treatment against aminoglycoside-induced hearing loss by co-administration of iron chelators has been successfully documented in guinea pig. This pharmacological intervention does not change serum levels of aminoglycosides nor their antibacterial efficacy. Since iron chelators are established therapeutic drugs, the proposed treatment should lend itself to clinical application. Aminoglycosides, long established for their high efficacy and low cost, may thus continue to play an important role in combating infectious diseases. PMID- 9339640 TI - ICD-10 and DSM-IV symptoms of somatoform disorders in different cultures. AB - In spite of an effort to harmonize the latest editions of International Classification of Diseases (ICD) and Diagnostic and Statistical Manual (DSM), a number of differences between the two classification systems still exist. One of these differences is related to the category of somatoform disorders. The cross cultural applicability of ICD-10 and DSM-IV criteria for somatoform disorders have been explored in the context of the ongoing WHO International Study of Somatoform Disorders. The study demonstrated that the current diagnostic concepts of somatoform disorders are cross-culturally acceptable in spite of the differences in their descriptions and classifications. However, it was found that a number of culture specific symptoms of somatoform disorders which do not appear in ICD-10 and DSM-IV were important and necessary for their diagnosis in specific cultures. PMID- 9339641 TI - Antitumor effects and pharmacological interaction of xiao-chai-hu-tang (sho-saiko to) and interleukin 2 in murine renal cell carcinoma. AB - Conventional therapy for renal cell carcinoma using interleukin 2 (IL-2) has shown limited antitumor action. The purpose of our study was to investigate synergistic antitumor effects of IL-2 and Xiao-Chai-Hu-Tang (XCHT), and to elucidate the mechanisms of interaction between the two drugs against the murine renal cell carcinoma cell line, Renca, in vivo. The treatment was started 5 days after subcutaneous transplantation of Renca tumor. XCHT was given at a dose of 2.5 g/kg daily for 30 days orally. IL-2 was given at a dose of 10(4) U/mouse by subcutaneous injection every other day 8 times. Combination of XCHT and IL-2 inhibited growth of the tumor and prolonged survival significantly as compared with the untreated mice. Increased cellular infiltration was observed in tumor tissue and the lungs of mice treated with XCHT alone and by combination of XCHT and IL-2, but there were no histological changes in the liver and kidney. Elevation of serum IL-6 was observed in tumor-bearing mice, but IL-6 was significantly suppressed by administration of XCHT. The results obtained suggest that combination of XCHT and IL-2 induces enhanced immunological reaction in specific organs and tissues, and IL-6 may have a role in the synergistic effect of these two agents. It was concluded that combination of XCHT and IL-2 is useful in the treatment of patients with renal cell carcinoma. PMID- 9339642 TI - A case of Klippel-Trenaunay syndrome associated with Huntington's disease. AB - We report here on a patient with Klippel-Trenaunay syndrome who was later diagnosed with Huntington's disease. Consistent with the later diagnosis, a (CAG)n repeat longer than the normal range was observed on chromosome 4p. The presence of these two diseases in the same individual may represent coincidence or a true correlation which must be confirmed by other evidence. To our knowledge, this is the first published report of the concurrent presence of these diseases in the same individual. PMID- 9339643 TI - Drug resistance studies using fresh human ovarian carcinoma and soft tissue sarcoma samples. AB - The relevance of continuous cell line cultures to the problem of clinical anticancer drug resistance is unclear. There is also mounting scepticism regarding the use of tumour cell lines with in vitro acquired drug resistance, possessing high levels of resistance unlikely to be seen in the clinical setting. To overcome some of these problems we have initiated a study of drug resistance using fresh tumour material obtained from patients suffering from ovarian cancer and soft tissue sarcoma (STS). Studies involving ovarian cancer have involved over 30 specimens of stage III-IV disease. For these samples we have specifically focused on the multidrug-resistant (MDR) phenotype, examining the role of proteins P-glycoprotein (Pgp), multidrug resistance-protein (MRP) and lung resistance-associated protein (LRP). Techniques have involved chemosensitivity testing, immunocytochemistry and flow cytometry, to measure Pgp function (drug efflux capacity with modulator reversal). Pgp was the most commonly expressed marker and its expression correlated with survival. MDR modulation using cyclosporins was shown to chemosensitise a proportion of the samples. Hence, in vitro screening can help to identify patients likely to benefit from resistance reversal strategies. Studies involving STS have looked at a combination of MDR and p53 disruption (commonly seen in this disease). Data have been examined alongside clinical data and the course of disease has been closely monitored. Although our studies are ongoing, we have identified a group of patients with aggressive disease showing marked drug resistance in vitro. All patients have relapsed with persistent disease following chemotherapy or radiotherapy. A number of chemoresistant patients showed a combination of p53 disruption in the presence of an MDR phenotype. Feedback from these translational studies should be used to guide the selection of patients for clinical trials using resistance reversal strategies and may suggest new targets for drug development. PMID- 9339644 TI - Malignant melanoma and interstitial pneumonia in a male. PMID- 9339645 TI - Early gastric cancer and lymph node metastasis. AB - A total of 1,845 cases of gastric cancer have been resected in our hospital during the past 18 years. Of these, 764 cases (41.5%) were early gastric cancer. The rate of lymph node metastasis was 2% in early mucosal cancer and 19.4% in early submucosal cancer (p < 0.001). In the cases of early mucosal cancer, the rate of lymph node metastasis was 0.9% for the elevated type and 2.4% for the depressed type. In the cases of early submucosal cancer, the rate of lymph node metastasis was 25.3% for the elevated type and 17.3% for the depressed type. In the mucosal cancers the rate of lymph node metastasis was high in the 0-IIc and IIc + III macroscopic type, and in the poorly differentiated microscopic type. In the submucosal cancers the rate of lymph node metastasis was 40% in mucinous adenocarcinoma and 33% in papillary adenocarcinoma. There was no lymph node metastasis in any case of early gastric cancer smaller than 1 cm surface diameter. PMID- 9339646 TI - Epidermal cell cultures from involved skin of patients with mammary and extramammary Paget's disease. AB - In involved epidermis, Paget cells are completely enclosed by the surrounding keratinocytes, which appear to be intact and unaltered. It is possible that the surrounding keratinocytes inhibit Paget cell proliferation. Accordingly, Paget cells might proliferate differently when cultured as an epidermal cell suspension. In this study, primary monolayer cultures of epithelial cells from involved epidermis of patients with mammary and extramammary Paget's disease were carried out to investigate whether Paget cells proliferate in the same manner as other malignant cells. Skin samples were obtained from one patient with mammary Paget's disease, and from 2 patients with extramammary Paget's disease. The epidermis was separated from the dermis with dispase, and epidermal cell suspensions were obtained with ethylenediamine tetraacetate and trypsin. A commercially available serum-free media, Keratinocyte-SFM, was used. Epithelial monolayers from the involved skin could be maintained for approximately 45 days, while keratinocytes from normal skin were maintained for approximately 35 days. The mechanism for the longer survival of the mixed cell culture of keratinocytes and Paget cells is not known. Permanent cell lines were not developed from these primary cultures. Paget cells could not be distinguished from keratinocytes by phase-contrast microscopy. The proportion of carcinoembryonic antigen (CEA) positive cells in the culture did not increase, but instead decreased. In certain areas of the dish, the CEA positive cells proliferated and accumulated like mushrooms. However, at the periphery of the dish, the Paget cells identified by immunostaining for CEA were dispersed and not clustered. These findings indicate that the influence of keratinocytes on Paget cells also occurs in cultured cells, which may explain why Paget cells survive longer than keratinocytes. In conclusion, the Paget cells in the involved epidermis do not proliferate like other malignant cells. PMID- 9339647 TI - Effects of interferon alpha 2a on incidence of hepatocellular carcinoma in chronic active hepatitis without cirrhosis. Hepatitis Treatment Study Group. AB - To determine the incidence of hepatocellular carcinoma among patients with chronic hepatitis C who received interferon (IFN) therapy, 63 patients with chronic hepatitis C who underwent IFN therapy (IFN alpha 2a 9 x 10(6) IU daily for 2 weeks and followed 9 x 10(6) IU three times weekly for 14 weeks) from January to December 1992, were studied. Selection criteria were as follows: within six months before IFN therapy patients were diagnosed with chronic active hepatitis without cirrhosis by hepatic histological examination, and were hepatitis C virus antibody positive. Furthermore, patients had records of follow up liver function tests (once a month) for more than six months after IFN therapy completion, and of ultrasound scanning (once in three to four months) before and for at least more than six months after the therapy completion. An average period of observation was 2.7 years (0.6 to 3.8 years). Twenty five of 63 patients (39.7%) returned to normal values of serum ALT, whereas 38 of 63 (60.3%) still showed abnormal values at six months after IFN therapy completion. Nine of 63 (14.2%) and 6/63 (9.5%) developed cirrhosis and hepatocellular carcinoma, respectively. All patients who developed cirrhosis and hepatocellular carcinoma were from those (n = 38) that showed abnormal ALT values after therapy completion. The five of six patients that progressed to hepatocellular carcinoma were associated with cirrhosis. No patients who returned to normal ALT values developed hepatocellular carcinoma during the period of observation. These results suggest that IFN therapy is effective to prevent the development of hepatocellular carcinoma. PMID- 9339648 TI - Gastric cancer during pregnancy. AB - During the 27 years between 1966 to 1993, a total of 3,256 patients with gastric cancer were admitted to our institute. Among these patients, 3 (0.1%) cases of gastric cancer were associated with pregnancy. One cancer stage IVb patient died of peritoneal metastasis without any operation. Another stage IVb patient died of peritoneal metastasis 12 months after palliative gastrectomy. And the other stage IIIb patient is well, and the 8 years old child is also well, after synchronous extended total gastrectomy with cesarean section. PMID- 9339649 TI - Expression of desmogleins in Paget cells of mammary and extramammary Paget's disease. AB - Sagebiel (1969) electronmicroscopically observed that desmosomes are present between adjacent Paget cells, as well as between Paget cells and adjacent keratinizing epidermal cells. Desmosomes contain the proteins desmoglein (Dsg) and desmocollin as their transmembrane components, and Dsg has three isotypes. Among the three isotypes of Dsg, Dsg1 and Dsg3 are autoantigens for pemphigus foliaceus and pemphigus vulgaris (PV), respectively. We examined the expression of Dsgs in Paget cells of mammary and extramammary Paget's disease. Skin samples were obtained from one patient with mammary Paget's disease, and from 2 with extramammary Paget's disease. One part of the samples was cut into small pieces and epidermal sheets were separated with dispase, then treated with a mixture of EDTA and trypsin. The resulting cell suspensions were cultured in low Ca++ medium, then the cells were incubated in high Ca++ medium. Paget cells identified with anti-epithelial membrane antigen (EMA) antibody were positive for serum from a PV patient which was confirmed to react with both Dsg1 and Dsg3. However, the intensity of fluorescence of Paget cells was weaker than that of EMA-negative keratinocytes. In the cryostat sections, Paget cells identified with anti-EMA antibody showed the same staining pattern as cultured cells in high Ca++ medium. The data presented in this study confirm that Paget cells express Dsgs, and are consistent with the electronmicroscopical data by Sagebiel (1969). However, our data do not support the hypothesis that Paget cells are of keratinocyte origin, because sweat ducts or sweat glands could be positive for sera from PV patients. It is necessary to confirm whether or not sweat ducts or sweat glands express Dsgs, because sera from PV patients exhibit high background in the dermis. PMID- 9339650 TI - Effects of endoscopic variceal ligation on systemic and splanchnic hemodynamics in patients with cirrhosis. AB - It has been shown that patients with cirrhosis and portal hypertension have hyperdynamic systemic and splanchnic circulation. This study was designed to assess how endoscopic variceal ligation may influence systemic and splanchnic hemodynamics. Sixteen patients with cirrhosis and esophageal varices were studied. Cardiac output and flow volume of the portal vein and the superior mesenteric artery were determined by means of duplex Doppler ultrasonography. Mean arterial pressure was also recorded. These hemodynamic measurements were performed before and after initial (3 days after initial session) and repeated (7 days after last session) variceal ligation. No significant changes in cardiac output and mean arterial pressure were found after either initial or repeated variceal ligation. Thus, systemic vascular resistance was not modified. In splanchnic hemodynamics, portal vein blood flow significantly increased after initial variceal ligation (27%, P < 0.01) but it returned to the baseline value after repeated variceal ligation. In contrast, superior mesenteric artery blood flow significantly decreased after initial variceal ligation (17%, P < 0.01) and it returned to the baseline value after repeated variceal ligation. Effect of variceal ligation on splanchnic hemodynamics is transient and variceal ligation has very little effect on systemic circulation. Thus, patients with cirrhosis underwent repeated variceal ligation still have abnormal systemic and splanchnic circulation. PMID- 9339651 TI - Ser-311-Cys polymorphism of the dopamine D2 receptor gene and schizophrenia--an analysis of schizophrenic patients in Fukuoka. AB - It has been suggested by Arinami et al. (1994) that there is a positive relationship between schizophrenia and the Cys311 polymorphism of the Dopamine receptor D2 (DRD2) gene. However, some recent reports do not support this relationship. The differing results could be due to two causes, [1] There may be a regional difference in the proportion of schizophrenia due to the Cys311 or [2] the control groups in the recent studies might have included young schizophrenics. Accordingly, an analysis of the age of schizophrenic patients in the Fukuoka region was undertaken. The Cys311 allele frequencies for the control group was 0.040 and for the schizophrenic group was 0.057, which were not statistically different. Although this data did not corroborate the above mentioned positive relationship, it was found that the allele frequency was higher statistically than in any previous report for both the schizophrenic and control groups. These findings indicate that the frequency of the Cys311 is biased geographically and that this must considered when investigating the occurrences of genetic variants and diseases. PMID- 9339652 TI - A histopathological study of pancreatic lesions after chronic administration of methamphetamine to rats. AB - The effects of chronic administration of methamphetamine on pancreatic tissues were histopathologically studied in experimental models. Methamphetamine (1 ml/kg body weight/day) was subcutaneously injected into 14 five-week-old male Wistar Kyoto rats (WKY) for 12 weeks. Age and sex matched 5 WKY rats served as controls. With light microscopy, some scattered edematous lesions and moderate vacuolization were demonstrated in the pancreas of 8 of the methamphetamine treated rats. However, in 4 of the rats, severe regional hemorrhage, partial acinal cell necrosis, destruction of the acinal cells, neutrophile infiltration, interstitial vessel dilatation, interstitial edema and fatty cell invasion were observed after the injections of methamphetamine. In 2 other animals, fibrosis and cirrhosis-like lesions with destruction and degeneration of the acinal cells were observed the small vessels had a slight degeneration of the endothelial cells. In the control animals, no lesions, except for some edematous lesions were found. In all cases, there were no nesidioblastosis-like lesions or necrosis of the Langerhans's islets. In the immunohistochemical study using anti- alpha 1 chymotrypsin antibody, more positive reactive cells were demonstrated among the interstitial and inter acinal cells, both in number and degree, in the methamphetamine treated rats. In addition to the animal model, there were 4 autopsy cases of sudden death in chronic methamphetamine abusers. The autopsies demonstrated a severe acute necrotic hemorrhagic pancreatitis, with only scattered slight hemorrhaging in the brain and lungs. These findings indicate that chronic administration of methamphetamine to rats evoked significant changes in pancreatic tissues including some degeneration of the endothelial cells of the small vessels in this hypoxia-vulnerable organ. PMID- 9339653 TI - Bone mineral density and the results of chiari pelvic osteotomy for osteoarthritis of the hip. AB - The aim of this study was to discover the involvement, if any, by bone mineral density (BMD) in the results after osteotomy for osteoarthritis of the hip (OA). The BMD, the Bombelli classification and the postoperative results in 36 joints after Chiari pelvic osteotomy for OA in the advanced and late stage were investigated for any correlations. The Bombelli hypertrophic type showed significantly better improvement than the atrophic type, but there was no correlation between the value of the BMD, and the postoperative result. PMID- 9339654 TI - Peripheral neuropathy in allergic granulomatous angiitis. AB - A 61-year-old woman began to suffer bronchial asthma in 1985. She then developed low back pain and numbness along the lower extremities, eventually leading to bilateral drop foot in 1990. At that time, she was diagnosed as having lumbar disc hernia, and extirpation of the discs at the L3-4 and L4-5 was performed. However, her clinical condition showed little improvement. Six months later, she was emaciated and bedridden with distal dominant muscular atrophy in all four limbs, purpura in the left leg and hypereosinophils. Motor conduction velocity (MCV) was not detected in the peroneal nerves. The toes gradually became cyanotic, and a skin biopsy from the cyanotic region revealed necrosis in the vessels surrounded by infiltration of a large number of neutrocytes and lymphocytes. She was diagnosed as having mononeuritis multiplex due to allergic granulomatous angiitis (AGA), which is characterized by bronchial asthma, hypereosinophilia and necrotizing vasculitis. Thirty mg/day prednisolone was then administered. However, the toes and calcaneal areas gradually became necrosed. Finally, amputation of both feet was necessary. We concluded that an early diagnosis of this syndrome is most important, and corticosteroids should be administered early. PMID- 9339655 TI - Stress fracture of the medial malleolus. AB - It is difficult to assess the healing of the medial malleolar stress fracture. We describe a case of medial malleolar stress fracture which was monitored its healing with repeated magnetic resonance images. PMID- 9339656 TI - An anomalous case of the gastro-hepatic and the spleno-mesenteric trunks independently arising from the abdominal aorta. PMID- 9339657 TI - [Ultrasonographic diagnosis of the parotid gland tumors--experience with 310 patients]. AB - The diagnostic value of ultrasonography of parotid gland tumors is to determine if the tumor is benign or malignant. In the period from 1984 to 1995, ultrasound examinations were performed on 310 patients with a space-occupying lesion of the parotid gland, using real time 3.75 MHz, 5 MHz and 7.5 MHz transducers. From a histological standpoint, there were 246 benign tumors, namely 144 pleomorphic adenomas, 35 adenolymphomas, 23 other adenomas, 14 cysts, 9 neurinomas, 3 lymphomas, 9 hemangiomas, 3 lymphangiomas and 6 lymph nodes. Another 64 were malignant tumors, namely, 7 mucoepidermoid carcinomas, 7 acinic cell carcinomas, 7 adenoid cystic carcinomas, 15 adenocarcinomas, 7 squamous cell carcinomas, 4 undifferentiated carcinomas, 10 carcinomas in pleomorphic adenoma, 3 malignant lymphomas, 2 metastatic carcinomas and 2 other carcinomas. According to our criteria for ultrasonographic diagnosis of the parotid gland tumors, the benign or malignant pattern was determined by the following findings; shape, boundary echo, internal echo and posterior echo. But in some cases we could not differentiate the two echogram patterns, and we call them the intermediate pattern. The total accuracy rate of the diagnosis of the 310 patients was 79.0%. (The intermediated pattern was found in 39 patients and these cases were considered to be misinterpreted.) In 268 primary parotid gland tumors (210 benign and 58 malignant tumors) excluding recurrent cases, lymph nodes, hemangiomas and so on, the sensitivity was 62.1%, the specificity was 91.4% and the total accuracy rate was 85.1%. The accuracy of sonography in determining whether a tumor was benign or malignant was affected by the tumor size; tumors smaller than 2 cm in diameter showed a tendency to be diagnosed as benign and tumors larger than 6 cm in diameter as malignant. In the 268 cases, the importance of the pattern of the boundary echo and the internal echo in differentiating benign and malignant tumors was examined retrospectively. In the 210 primary benign tumors, the benignity of the tumors was assessed correctly in 84.3% by the boundary echo and 85.7% by the internal echo. In the 58 primary malignant tumors, the malignancy was assessed correctly in 58.6% by the boundary echo and 43.1% by the internal echo. Strong echos, which are the signs of a malignant pattern, were seen in 14.8% of the benign tumors, and in 51.7% of the malignant tumors. It can be concluded that ultrasonography is very reliable for the examination of parotid gland tumors. PMID- 9339658 TI - [Scanning and transmission electron microscopic observations of the inner ear of hamsters with hyperlipidemia]. AB - The relationship between hyperlipoproteinemia and sensorineural hearing loss has been studied by means of guinea pig models with hypercholesterolemia. However, these observations of the inner ear using guinea pig models have been limited to a short time. By using a golden hamster model with hypercholes terolemia, I was able to observe the inner ear for a long time. Two-month-old hamsters were fed a hyperlipid diet consisting of standard chow supplemented with 3% cholesterol and 15% cattle fat for 30, 60, 90, 120, and over 150 days. Six-month-old hamsters were fed the hyperlipid diet for 30 days. Then the animals were examined for auditory dysfunction and morphological changes in the cochlea. Biochemical findings in the serum showed hyperlipoproteinemia, especially hypercholesterolemia. Regarding auditory dysfunction, the threshold change in the auditory brainstem response (ABR) was mild. Scanning and transmission electron microscopic revealed many protrusions toward the endolymphatic space on the surface of the marginal cells of the stria vascularis. Vascular degeneration of the marginal cells and intermediate cells of the stria vascularis was also observed. In addition, protrusions containing a lysozome structure consisting of outer piller cells were observed on the organ of Corti. These results with experimental hamsters fed a hyperlipid diet indicate that such a diet may induce functional changes in the cochlea such as auditory dysfunction with the occurrence in hypercholesterolemia. PMID- 9339659 TI - [Quantitative analysis of the gustatory evoked potentials in the guinea pig]. AB - Gustatory evoked potentials were studied in anesthetized guinea pigs to develop an objective and quantitative taste examination for patients with taste disorders. A positive wave was recorded by the application of NaCl, HCl or quinine hydrochloride solution. There was little difference in latency, duration and waveform among these three solutions. No apparent change in activity was seen after the application of sucrose solution or distilled water. The gustatory evoked potentials that excluded the influence of the trigeminal nerve innervating the tongue surface were able to be reproducibly recorded on either the cortical surface or the skull surface. There was a linear relationship between logarithmic values of potential amplitude and those of taste solution concentration. Therefore, it is suspected that the quantitative evaluation of taste detection is possible by measuring the taste solution concentration-potential amplitude relationship. PMID- 9339660 TI - [The serum zinc level in patients with tinnitus and the effect of zinc treatment]. AB - We measured the serum zinc level in patients with tinnitus and evaluated the effectiveness of zinc in the treatment of tinnitus. Blood zinc levels were measured in 121 patients with tinnitus. All patients were examined between 1995 and 1997 at the outpatient clinic of otorhinolaryngology St. Marianna University Toyoko Hospital. Forty-seven patients who had received any drug such as a calcium channel blocker and others or had been affected by any diseases were excluded and therefore 74 patients consisting of 46 females (62%) and 28 males (38%) were investigated. Twenty two healthy volunteers served as a control group. The mean age and standard deviations for the tinnitus group and the control group were 47.8 +/- 17.1 and 31.4 +/- 8.2 years, respectively. There was a significant decrease (p < 0.0001) in serum zinc levels in patients with tinnitus compared with the control group. Because there was a significant difference (p < 0.0001) in age distribution between tinnitus and control groups, patients were selected by their age in order to neglect the effect of aging. In this situation, a significant difference (p < 0.01) was noted between the tinnitus group and control group. Low blood zinc level was defined by using the mean and standard deviation for the control group (mean-1 S.D.). We treated patients with low blood zinc levels. A total dose of 34-68 mg of Zn++ was administered daily for over 2 weeks. The degree of tinnitus was expressed on a numeric scale from 0 to 10 before and after treatment. Blood zinc levels were significantly elevated (p < 0.05) after treatment. We found a significant decrease (p < 0.01) in the numeric scale. These findings suggest that zinc is useful in at least some patients suffering from tinnitus. It is possible to classify patients with tinnitus by measuring serum zinc level and this leads to improvement of the overall treatment effect. PMID- 9339661 TI - [Suppression of tinnitus by band noise masker--a study of 600 cases]. AB - We performed Band Noise Masker (BNM) therapy for the suppression of tinnitus in 600 patients and measured the pitch, loudness and masking level of tinnitus and residual inhibition (RI). We examined the efficiency of BNM therapy. The purpose of this study was to investigate the mechanism of suppression of tinnitus by BNM. Tinnitus was suppressed in 394 patients (66%) after BNM therapy. In the group of patients in whom we suppressed tinnitus, the loudness of tinnitus was reduced from 7.7 +/- 5.7dBSL to 7.5 +/- 5.5dBSL (p < 0.05) and the pitch of tinnitus did not exhibit a marked change. In the group of patients in whom we did not suppress tinnitus, the loudness and pitch of tinnitus did not exhibit a marked change. The efficiency of BNM therapy was high in the cases of presbyacusis and low in the cases of sudden deafness. There was no significant relationship between RI and the efficiency of BNM therapy. We examined these data and discussed the mechanism of suppression of tinnitus by BNM therapy. In all cases, after BNM therapy, the auditory threshold did not become worse. In 4 cases tinnitus became worse temporarily. In conclusion, BNM therapy is an effective mode for tinnitus control, is easily performed in our outpatient clinic or at home, and has no serious complications. PMID- 9339662 TI - [Formation and calcium incorporation of giant otoconia of the guinea pig after streptomycin intoxication]. AB - The mechanisms for the formation and fate of giant otoconia following streptomycin (SM) intoxication were investigated in adult pigmented guinea pigs by scanning electron microscopy. Calcium turnover into otoconia has also been studied by using tetracycline as a tracer. The administration of SM induced the reduction of otoconia with the formation of giant otoconia. The giant otoconia had a multifaceted morphology in their early developmental period. This type of otoconia showed entire fluorescence indicating existence of calcium uptake. They then grew up to the transitional type and finally to the cylindrical type. The giant otoconia were thought to be formed mainly by dissolution of normal otoconia due to the loss of environmental calcium followed by recrystallization as giant crystals. The transitional type of giant otoconia showed less calcium ion uptake and the removal of calcium from the giant otoconia caused their quick disappearance. These phenomena might be closely related to the otoconial dynamics which may regulate calcium ion homeostasis of endolymph. PMID- 9339663 TI - [Examination of low palatal arch with long low hanging soft palate in obstructive sleep apnea syndrome and cephalometry]. AB - In order to clarify morphological abnormalities of the upper airway in patients with obstructive sleep apnea syndrome (OSAS), and to predict the severity as well as the effect of surgery, we statistically examined the relationship between the degree of low palatal arch and the result of cephalometry in 45 adult male patients with OSAS who underwent uvulopalatopharyngoplasty. The patients were divided into 3 types, A, B and C, according to the degree of low palatal arch on inspection, and type C was regarded as having a low palatal arch with a long low hanging soft palate. It was found that (1) type C patients showed a higher preoperative apnea index and lower minimal blood oxygen saturation, suggesting a serious condition. (2) In type C patients, the long axis (PNS-H) of the airway was larger, and the oral area was smaller. The area of the entire tongue, especially the upper half of the tongue, was larger than that in patients of other types. Type C patients were further divided into 3 subtypes: (1) those with a substantially long low-hanging soft palate; (2) those with a long low-hanging soft palate and large tongue; and (3) those in whom the large tongue is the main factor for the apparent low-hanging soft palate. These subtypes should be confirmed by cephalography before treatment for effective surgical results. PMID- 9339664 TI - [Unilateral multiple tumorous lesions of the parotid gland]. AB - Multifocal tumors within the same parotid gland are very rare. We treated 13 patients with multiple tumorous lesions within the unilateral parotid gland. We evaluated the multiple nodules by CT-sialography or magnetic resonance imaging (MRI). These imagings showed clearly two or more distinct nodular-appearing lesions. Recurrent pleomorphic adenoma (6 patients) was predominant, followed by Whartin's tumor (3 patients). The other lesions were two differential parenchymal tumors (polymorphous low grade adenoma/adenoma) within the same gland, a malignant lymphoma, a squamous cell carcinoma metastatic to the gland, and a tuberculous lesion. On palpitation, 9 of the patients had an unilateral tumor, one a palpable parotid mass in the gland, and the other four had two or more tumors in the unilateral gland. The patients with intra-parotid lymph node and metastatic lesions had extra-parotid cervical adenopathy. The clinical features and the differential diagnosis of the unilateral multiple tumorous lesions of the parotid gland are discussed. PMID- 9339665 TI - New techniques for investigating mitochondrial DNA in neurodegenerative diseases. PMID- 9339666 TI - Synchrony, sleep, dreams, and consciousness: clues from K-complexes. PMID- 9339667 TI - Challenges in the design and conduct of phase III brain tumor therapy trials. PMID- 9339668 TI - Cybrids in Alzheimer's disease: a cellular model of the disease? AB - The mitochondrial electron transport chain enzyme cytochrome c oxidase (COX) is defective in patients with sporadic Alzheimer's disease (AD). This defect arises from the mutation of mitochondrial DNA (mtDNA). To develop a tissue culture system that would express this genetically derived bioenergetic lesion and permit characterization of its functional consequences, we depleted Ntera2/D1 (NT2) teratocarcinoma cells of endogenous mtDNA and repopulated them with platelet mtDNA from AD patients. Cytochrome c oxidase activity was depressed in the resulting AD cytoplasmic hybrids (cybrids) compared with cybrids prepared with mtDNA from non-AD controls. Reactive oxygen species (ROS) production and free radical scavenging enzyme activities were significantly elevated in AD cybrids. A COX defect in NT2 AD cybrid lines indicates that AD patients possess mtDNA COX gene mutations that are sufficient for determining this biochemical lesion. Expression of unique functional characteristics (increased ROS production and free radical scavenging enzyme activities) relevant to neurodegeneration demonstrates the utility of these cells in defining AD pathophysiology at a cellular level. This in vitro tissue culture model of AD may prove useful in drug screening. PMID- 9339669 TI - Evaluation of dementia: a systematic study of the usefulness of the American Academy of Neurology's practice parameters. AB - OBJECTIVE: This study examined the usefulness of the "Practice Parameters for the Evaluation of Dementia," published by the Quality Standards Subcommittee of the American Academy of Neurology (1994). The Practice Parameters are stratified according to three levels of certainty (standards, guidelines, and options), and suggest indications for the use of neuroimaging studies. METHODS: We reviewed 119 consecutive cases referred for assessment of memory loss to a university affiliated interdisciplinary clinic. We assessed the diagnostic value of laboratory, neuropsychological, and neuroimaging studies above the standard history, neurologic, and mental status examinations. We also assessed the sensitivity and specificity of four clinical indicators (i.e., early symptom onset, noninsidious course, focal neurologic signs or symptoms, and gait disturbance) for predicting the diagnostic utility of neuroimaging studies. RESULTS: The largest changes in diagnostic categories between the standard and the comprehensive diagnostic process was a 9% decrease in the diagnosis of Alzheimer's disease, a 6% increase in the diagnosis of mixed dementia (due largely to laboratory studies), and a 4% increase in the diagnosis of vascular dementia (due to neuroimaging). The clinical indicators were 82% sensitive and 50% specific in predicting that neuroimaging studies would change the diagnosis. In six cases, meaningful neuroimaging findings were not associated with any clinical indicator (5% false negatives). In 43 cases, neuroimaging provided no significant clinical findings despite the presence of an indicator (36% false positives). CONCLUSIONS: In this convenience sample, diagnostic accuracy was improved to a comparable degree by laboratory and neuroimaging studies, although at a significant difference in cost. Use of the four clinical indicators would have reduced the frequency of neuroimaging studies by 33% but missed clinically meaningful information in 5%. Although imperfect, the Practice Parameters represent a first step toward improving the cost effectiveness of the dementia work-up. PMID- 9339670 TI - Search patterns using the line bisection test for neglect. AB - BACKGROUND/OBJECTIVE: Neglect has been attributed to sensory-attentional, motor intentional, and representational disorders. Although patients with neglect may have defective visual exploration, the nature of this exploration deficit has not been entirely elucidated. The purpose of this study is to learn how, compared with control subjects, a patient with neglect performs visual exploration. METHODS: The line bisection test is commonly used to detect and evaluate the performance of patients with unilateral or hemispatial spatial neglect. We tested an experimental subject with left premotor-intentional neglect using an infrared eye monitoring instrument. Thirteen right-handed healthy volunteers served as control subjects. From different starting ocular positions, we assessed the direction of eye movements when the subjects initially oriented to the line, scanned the line, and then attempted to look at the center of the line. RESULTS: Independent of starting ocular position, normal subjects most often initially orient to the left end of the line, scan rightward, then look leftward to the center. However, the eye movements in our experimental subject with neglect were directed to the right end of the line without any scanning or leftward eye movement back to the center of the line. CONCLUSION: Our subject with neglect had defective line exploration that may be attributed to a directional ocular motor intentional deficit. PMID- 9339671 TI - Striatal dopamine metabolism correlated with frontotemporal glucose utilization in Alzheimer's disease: a double-tracer PET study. AB - We previously found that the rate of [l8F]6-fluoro-L-dopa (FDOPA) uptake into the striatum (the Ki value) in Alzheimer's disease (AD) correlated with cognitive function. Patients with wandering behavior had an increased level of D2 dopamine receptors. Decreased cerebral glucose utilization (CMRglc) in the parietal and temporo-parieto-occipital region is a well-known PET finding in AD. We investigated the relationship between the Ki value and regional CMRglc (rCMRglc) with reference to cognitive impairment and wandering behavior. We studied 10 AD patients with moderately severe dementia. Using PET and the FDOPA and [18F] fluoro-deoxyglucose techniques, the Ki value and rCMRglc in 10 regions in each hemisphere were measured. The Ki value was found to correlate with mean cortical rCMRglc. The rCMRglc of the anterior frontal, inferior frontal, and temporal lobes correlated with the Ki value; the patients with wandering behavior had lower values. Multiple regression analysis showed that the Mini-Mental State Examination score was significantly explained by the Ki value, the rCMRglc of the temporal lobe, and the rCMRglc of the parietal lobe. There is probably a functional neural network between the striatum and the frontal and temporal lobes in AD. We consider this network important, not only in cognitive impairment but also in abnormal wandering behavior. PMID- 9339672 TI - Frontal pure agraphia for kanji or kana: dissociation between morphology and phonology. AB - We present two patients with frontal pure agraphia more impaired for either kanji or kana (two separate writing systems for the Japanese language). The lesion of patient 1 (preferentially disturbed for kanji) was restricted to the foot of the middle frontal gyrus and the adjacent anterior precentral gyrus, whereas the lesion of patient 2 (preferentially disturbed for kana) included the posterior two thirds of the middle frontal gyrus. Both patients made agraphic errors (impaired recall) for kanji and agraphic or paragraphic errors (changing into other symbols) for kana. The double dissociation and the difference in types of errors between kanji writing and kana writing suggests that there are two pathways involved in writing, i.e., a morphologic route and a phonologic route. We concluded that damage to the morphologic route may yield agraphia for kanji and that damage to the phonologic route may yield agraphia for kana. PMID- 9339673 TI - The K-complex: its slow (<1-Hz) rhythmicity and relation to delta waves. AB - The K-complex is a major graphoelement of sleep EEG. This report demonstrates that K-complexes emerge from a cortically generated slow (<1-Hz) oscillation. Human EEG as well as cat cellular and field potential recordings converge into demonstrating that the K-complex results from a synchronized cortical network that imposes periodic excitatory and inhibitory actions on cortical neurons. We additionally show the correspondence between neuronal activities and the shape of the K-complex. Spectral analysis confirms the periodic recurrence of human K complexes, with main peaks at 0.5 to 0.7 Hz. It is also shown that the spectral content in the delta band (1 to 4 Hz) is partially due to the shape and duration of the K-complex. PMID- 9339674 TI - Temporal lobe epilepsy in childhood: clinical, EEG, and neuroimaging findings and syndrome classification in a cohort with new-onset seizures. AB - Sixty-three children with new-onset temporal lobe epilepsy (TLE) underwent extensive clinical, EEG, and neuroimaging investigation as part of a prospective, community-based cohort study of the natural history of TLE in childhood. Complex partial seizures occurred in 94% of the children, and tonic-clonic seizures occurred in 14%. Developmental, behavioral, or learning problems were present in 38%. Eighteen children (29%) had a significant illness/event prior to the onset of TLE, including febrile status epilepticus in seven, meningitis in four, respiratory arrest in two, and head injury in one. Magnetic resonance imaging or computed tomography revealed structural abnormalities of the temporal lobe in 24 children (38%), including hippocampal sclerosis (HS) in 13 and tumor in eight. There was a strong association between HS and a history of significant illness/event prior to the onset of TLE (p < 0.001). Analysis of past history and neuroimaging findings led us to propose three etiologically defined subgroups of TLE; developmental TLE (10 children with long-standing, nonprogressive temporal lobe tumors and malformations), TLE with HS/significant antecedents (18 children with HS or a history of a significant illness/event), and cryptogenic TLE (34 children with normal neuroimaging findings and no significant past history). Etiologic differences between children with new-onset TLE may confer prognostic information that will be useful for counselling families and planning treatment. PMID- 9339675 TI - Autosomal dominant nocturnal frontal lobe epilepsy: demonstration of focal frontal onset and intrafamilial variation. AB - Autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) is a newly recognized autosomal dominant partial epilepsy. We studied seizure localization and intrafamilial variation using video-EEG monitoring (VEM) and functional neuroimaging in two pairs of subjects from unrelated families. The clinical features of seizures were similar from seizure to seizure in each individual, but varied between individuals. As is often found in frontal lobe epilepsies, ictal EEG localization was imprecise in three of four cases. One patient showed a consistent left fronto-polar onset that was corroborated by congruent focal hypometabolism on interictal PET and focal hyperperfusion on ictal single photon emission computed tomography (SPECT). A second case studied with ictal SPECT showed a right parasagittal, midfrontal focus. We conclude that this autosomal dominant epilepsy syndrome, which in one of the two families was due to a known neuronal nicotinic acetylcholine receptor mutation, causes frontal lobe foci that are unilateral and in variable locations in different individuals. PMID- 9339676 TI - Mesial temporal spikes interfere with working memory. AB - The hippocampus and other mesial temporal structures support long-term memory and also are common foci for epilepsy. Recently it was shown that these brain structures may subserve the short-term storage and rehearsal processes called working memory in humans. We determined the accuracy of verbal and visuospatial working memory in the presence and in the absence of mesial temporal spikes in eight patients who had bilateral depth electrodes implanted to evaluate intractable epilepsy. Six of eight patients had declines in working memory performance during mesial temporal spiking, with the greatest disruption in spatial and verbal recall coincident with left hippocampal spikes (p = 0.019). Overall accuracy of working memory for all patients declined an average of 6% on spike trials. The two patients who did not have decreased accuracy during spike trials also had the best overall working memory performance. Mesial temporal spikes were not detected on extracranial recordings and yet may be associated with declines in working memory in some patients with epilepsy. PMID- 9339678 TI - Treatment of first tonic-clonic seizure does not improve the prognosis of epilepsy. First Seizure Trial Group (FIRST Group). AB - It is widely agreed that after two or more seizures patients should be given antiepileptic treatment, but there is still controversy about the treatment of patients after a first unprovoked seizure. In a multicenter, randomized, open trial, patients with a first tonic-clonic seizure were randomized to immediate treatment (carbamazepine, phenytoin, phenobarbital, or sodium valproate) or to treatment only after another seizure. Fifty-two (24%) of the 215 patients randomized to immediate treatment and 85 (42%) of the 204 randomized to delayed treatment experienced seizure recurrence during follow-up. Age, acute treatment of the seizure with benzodiazepines, remote etiologic factors, and EEG abnormalities were significant predictors of relapse. Of the immediately treated patients, 87% had no seizures for a year and 68% had no seizures for 2 years, whereas only slightly fewer initially untreated patients (83% and 60%) achieved these endpoints. Patients treated after the first seizure and those treated after seizure relapse had the same time-dependent probability of achieving 1 and 2 seizure-free years. None of the variables that were prognostic predictors of relapse was significantly associated with the probability of having 1 or 2 years of seizure control. Anticonvulsants in patients presenting a first tonic-clonic seizure reduce the risk of relapse; however, 50% of patients who are not treated will never experience a second seizure. Moreover, the probability of long-term remission is not influenced by treatment of the first seizure. PMID- 9339677 TI - Single photon emission computed tomography-EEG relations in temporal lobe epilepsy. AB - The role of single photon emission computed tomography (SPECT) as an independent confirmation test in presurgical evaluation of medically intractable temporal lobe epilepsy has not been critically investigated. Because spreading ictal discharges may cause a concomitant increase of cerebral blood flow in remote cerebral regions, a careful analysis of peri-injection EEG patterns and their relation to ictal SPECT may be important in evaluating the reliability of ictal SPECT. Both interictal and ictal EEG and SPECT were reviewed in 19 patients with temporal lobe epilepsy who achieved a successful seizure outcome after surgery. Patients were divided into unitemporal and bitemporal groups according to the lateralization of interictal epileptiform discharges (IED). Ictal EEG features were classified into lateralized and nonlateralized groups. The concordance between SPECT and EEG lateralizations was examined in each patient and correlated to the documented epileptogenic temporal lobe. Interictal SPECT correctly lateralized in eight of nine patients with unitemporal IED and in five of 10 patients with bitemporal IED. Ictal SPECT was highly concordant with the peri injection ictal EEG but correctly lateralized the epileptogenic region in only 11 of 19 patients. When both pre- and postinjection EEG epochs lateralized ipsilaterally, all ictal SPECT images showed concordant lateralization. If pre- and postinjection EEG epochs were either different in lateralization or nonlateralization, ictal SPECT images often showed complex patterns of cerebral perfusion with a high incidence of false lateralization. Interictal SPECT was more sensitive and reliable in patients with unitemporal IED than in patients with bitemporal IEDs. Ictal SPECT was closely related with peri-injection EEG epochs but with frequent false lateralization. The role of ictal SPECT as an independent confirmation test in presurgical evaluation should be reappraised. PMID- 9339679 TI - Sleep and quantitative EEG in patients with progressive supranuclear palsy. AB - Sleep architecture and quantitative EEG from wakefulness and REM sleep were studied in six patients (mean age, 70.5 years) with progressive supranuclear palsy (PSP) and compared with that of six control subjects (mean age, 69.8 years). Particular attention was given to quantifying REM sleep variables because of the known PSP-associated degeneration of the pedunculopontine tegmentum (PPT)- a critical structure in REM sleep generation. Patients with PSP had a shorter total sleep time, a lower sleep efficiency, a drastic reduction in sleep spindles, an atonic slow-wave sleep, and a lower percentage of REM sleep. The lower percentage of REM sleep was the result of both a reduction in the number of REM periods and a reduction in mean period of duration. REM density was also reduced while REM efficiency, atonia, and phasic EMG were similar to control values. REM sleep findings are consistent with the known role of the PPT in REM sleep induction. A slowing of the awake EEG was found for the six frontal leads and for C4, P4, and T4 in PSP patients. The frontal EEG slowing found in wakefulness is in accord with imaging and neuropsychological studies showing impairment of the frontal lobes in these patients. REM sleep EEG was not significantly slower in any regions. Because all previous studies on PSP have relied on visual inspection of the EEG tracings, the present finding of EEG slowing in the frontal lobes (rather than in the temporal regions or diffusely) suggests that our quantitative EEG approach may be more useful in determining specific regions of impaired cortical activity. PMID- 9339680 TI - Friedreich's ataxia GAA repeat expansion in patients with recessive or sporadic ataxia. AB - To explore the clinical heterogeneity associated with the Friedreich's ataxia (FRDA) expanded repeat and provide preliminary guidance for future gene testing in patients suspected of having FRDA, we tested patients with typical FRDA (group I), late-onset FRDA or FRDA with retained reflexes (group II), as well as those with early onset "non-Friedreich's" recessive or sporadic ataxia (group III). Eighty-seven percent of families in group I tested positive for the FRDA triplet repeat expansion. Thirty-six percent of families in group II demonstrated the FRDA expansion. Only one of 11 patients in group III had the FRDA expansion. Clinical criteria did not clearly distinguish between expansion-positive and expansion-negative individuals in groups I and II. Minimal criteria that were present in all the patients who tested positive were recessive or sporadic inheritance, progressive caudal-rostral gait and limb ataxia, and at least one of the following: dysarthria, Babinski sign, or cardiomyopathy. This study confirms recent findings that some patients in group II can carry the FRDA mutation. However, we did not observe the FRDA expansion in 64% of group II families or in 13% of families with typical FRDA (group I), suggesting other genetic or environmental causes for their ataxia. PMID- 9339681 TI - The expansion of the CAG repeat in ataxin-2 is a frequent cause of autosomal dominant spinocerebellar ataxia. AB - The autosomal dominant spinocerebellar ataxias (ADSCAs) are a heterogeneous group of late-onset neurodegenerative disorders with overlapping clinical features. Genetic linkage studies have identified at least seven distinct loci for the ADSCAs, allowing the genetic classification of these disorders. The spinocerebellar ataxia type 2 (SCA2) locus was mapped to chromosome 12, and a gene responsible for this disorder was recently isolated. The mutation causing SCA2 is an expansion of a trinucleotide CAG repeat contained within the coding region of a novel gene. We describe the results of genotypic analysis for the SCA2 repeat in individuals with ADSCA who were previously found negative for CAG repeat expansions in the SCA1, SCA3, or SCA6 genes. The expanded CAG repeat has been identified in 15 independent families. Repeat instability and anticipation were observed in two large kindreds. The SCA2 mutation was found in 18% of our ADSCA kindreds, confirming the high proportion of SCA2 among this group of disorders. PMID- 9339682 TI - Pigmentary retinopathy associated with the mitochondrial DNA 3243 point mutation. AB - Fourteen patients from four unrelated families were studied to determine the prevalence of retinal pigmentary abnormalities associated with the MELAS A to G 3243 point mutation. Neurologic and ophthalmic examinations, retinal photography, pattern shift visual evoked potentials, and electroretinography were performed in all patients. Eight of the 14 patients had retinal pigmentary abnormalities characterized by symmetric areas of depigmentation involving predominantly the posterior pole and midperipheral retina. None of the patients had optic atrophy and only one patient with pigmentary retinal abnormalities had impaired visual acuity. None of the diabetic subjects (n = 6) had signs of diabetic retinopathy. Fluorescein angiography demonstrated mottled hyper- and hypofluorescent areas indicating multiple window defects in the retinal pigmentary epithelium. Visual evoked potentials showed delayed P100 responses in four of the eight patients with retinal pigmentary abnormalities. We conclude that there is a high prevalence of retinal pigmentary abnormalities in patients with MELAS A to G 3243 point mutation. These abnormalities are usually asymptomatic and best detected by retinal photography. PMID- 9339683 TI - A novel sodium channel mutation causing a hyperkalemic paralytic and paramyotonic syndrome with variable clinical expressivity. AB - A point mutation A4078G predicting the amino acid exchange Met1360Val in segment IV/S1 of the human muscle sodium channel alpha-subunit was identified in a family presenting features of hyperkalemic periodic paralysis and paramyotonia congenita with sex-related modification of expression. In this family, only one male member is clinically affected, presenting episodes of flaccid weakness as well as paradoxical myotonia and cold-induced weakness. Three female family members who have the same mutation show only myotonic bursts on EMG. We studied the functional defect caused by this mutation by investigating recombinant wild type (WT) and mutant sodium channels expressed in a mammalian cell line (HEK293) using the patch-clamp technique. With mutant channels, the decay of the sodium currents was two times slower than with WT, the steady-state inactivation curve was shifted by -13 mV, and recovery from inactivation was 1.5 times faster. High extracellular potassium (9 mM) did not affect channel gating. Single-channel measurements revealed prolonged mean open times and an increased number of channel reopenings. The results are remarkable with respect to the lack of complete penetrance usually seen with sodium channelopathies and the site of mutation that was formerly not thought to be involved in channel inactivation. PMID- 9339684 TI - Autoantibodies to glutamic acid decarboxylase in three patients with cerebellar ataxia, late-onset insulin-dependent diabetes mellitus, and polyendocrine autoimmunity. AB - BACKGROUND: Glutamic acid decarboxylase (GAD) is the main target of humoral autoimmunity in stiff-man syndrome (SMS) and insulin-dependent diabetes mellitus (IDDM). GAD autoantibodies (GAD-Abs) are reported in a few patients with cerebellar ataxia, but their relevance is unclear. We describe three patients with cerebellar ataxia and GAD-Abs. METHODS: GAD-Abs were assayed by radioimmunoassay (RIA) and immunohistochemistry and confirmed by immunoblot of recombinant human GAD65. The GAD-Ab levels of the three patients with cerebellar ataxia were compared with those of five with SMS, 49 with IDDM, 64 with cerebellar ataxia of probable degenerative origin without associated autoimmune features, 14 non-IDDM islet cell antibody-positive first-degree relatives of IDDM patients, and 91 normal subjects. RESULTS: The three patients with ataxia and GAD Abs were women (mean age, 63 years) with an isolated progressive cerebellar disorder, family history of IDDM, late-onset IDDM, and several positive serum organ-specific autoantibodies. Two patients had autoimmune thyroiditis, and one had pernicious anemia. CSF analysis demonstrated oligoclonal IgG bands and intrathecal synthesis of GAD-Abs. By RIA, GAD-Ab titers from the three patients were similar to those of SMS and significantly higher, without overlap, than the titers of IDDM patients. GAD-Abs were absent in the 64 patients with cerebellar ataxia and no evidence of autoimmune disorders. CONCLUSIONS: These findings suggest a link of GAD autoimmunity not only with SMS but also with cerebellar dysfunction. GAD-Abs should be sought in patients with cerebellar ataxia who have late-onset IDDM and other organ-specific autoimmune manifestations. PMID- 9339686 TI - Higher neonatal cerebral blood flow correlates with worse childhood neurologic outcome. AB - Cerebral blood flow (CBF) in newborn infants is often below levels necessary to sustain brain viability in adults. Controversy exists regarding the effects of such low CBF on subsequent neurologic function. We determined the current childhood neurologic status and IQ in 26 subjects who had measurements of CBF performed with PET in the neonatal period between 1983 and 1989 as part of a study of hypoxic-ischemic encephalopathy. Follow-up information at ages 4 to 12 years was obtained on all 26 subjects. Ten subjects had died. All 16 survivors underwent clinical neurologic evaluation, and 14 also underwent intelligence testing. Eight had abnormal clinical neurologic evaluations; eight were normal. The mean neonatal CBF in those with abnormal childhood neurologic outcome was significantly higher than in those with normal childhood neurologic outcome (35.64 +/- 11.80 versus 18.26 +/- 8.62 mL 100 g(-1) min(-1), t = 3.36, p = 0.005). A significant negative correlation between neonatal CBF and childhood IQ was demonstrated (Spearman rank correlation r = -0.675, p = 0.008). Higher CBF was associated with lower IQ. The higher CBF in subjects with worse neurologic and intellectual outcome may reflect greater loss of cerebrovascular autoregulation or other vascular regulatory mechanisms due to more severe brain damage. PMID- 9339685 TI - Autonomic failure in Guamanian neurodegenerative disease. AB - Autonomic impairment is minor in idiopathic amyotrophic lateral sclerosis (ALS) and Alzheimer's-type dementia (D) and is usually not marked in Parkinson's disease. The autonomic status of Guamanian parkinsonism (P), ALS, and parkinsonism-dementia complex (PDC) is essentially unknown. We therefore evaluated the autonomic nervous system in Guamanian D, ALS, P, and PDC. Cardiovagal, adrenergic, and postganglionic sudomotor functions were quantitated in 16 patients and 14 paired household controls. Patients comprised PDC (N = 11), D (N = 2), P (N = 2), and ALS (N = 1). Autonomic deficit was expressed on a composite autonomic scoring scale (CASS) and its subsets that corrects for the effects of age and gender. CASS severity was rated from 0 to 10 and the maximal subset scores were 3, 3, and 4 for postganglionic sudomotor, cardiovagal, and adrenergic deficits, respectively. CASS scores for mild, moderate, and severe autonomic failure are 1 to 3, 4 to 6, and 7 to 10, respectively. Symptoms were scored by an Autonomic Symptom Profile (ASP). The affected patients were older than and had a sex distribution different from paired controls (64.2 +/- 8.0 versus 53.1 +/- 13.5; p < 0.01; male/female = 9/7 versus 2/12; p = 0.045). CASS scores were markedly increased over paired controls (6.2 +/- 2.3 versus 1.9 +/- 1.3; p < 0.001), and involvement was generalized by system. There were deficits in sudomotor, cardiovagal, and adrenergic function. Orthostatic hypotension occurred in 6 of 16 patients and 2 of 14 paired controls. Guamanian patients had more autonomic dysfunction than non-Guamanian Parkinson's disease. ASP scores were higher in patients than controls and regressed with CASS. These differences persisted when corrected for the confounding effects of age, gender, and diabetes. We conclude that Guamanian patients have autonomic failure to a greater extent than non-Guamanian Parkinson's disease or ALS. This autonomic failure suggests multisystem autonomic involvement similar to but less severe than in multiple system atrophy. PMID- 9339687 TI - Bilateral periventricular nodular heterotopia with mental retardation and syndactyly in boys: a new X-linked mental retardation syndrome. AB - Bilateral periventricular nodular heterotopia (BPNH) is a recently recognized malformation of neuronal migration, and perhaps proliferation, in which nodular masses of gray matter line the walls of the lateral ventricles. Most affected individuals have epilepsy and normal intelligence with no other congenital anomalies. A striking skew of the sex ratio has been observed because 31 of 38 probands have been female, and one gene associated with BPNH was recently mapped to chromosome Xq28. We report three unrelated boys with a new multiple congenital anomaly-mental retardation syndrome that consists of BPNH, cerebellar hypoplasia, severe mental retardation, epilepsy, and syndactyly. Variable abnormalities included focal or regional cortical dysplasia, cataracts, and hypospadius. We hypothesize that this syndrome involves the same Xq28 locus as isolated BPNH, and we review the expanding number of syndromes associated with BPNH. PMID- 9339688 TI - Genetic testing of children at risk for Huntington's disease. US Huntington Disease Genetic Testing Group. AB - We reviewed 44 symptomatic children tested for CAG repeat expansions in the gene responsible for Huntington's disease (HD). Thirty-three patients had CAG repeat expansions, and 11 did not. No patient with a CAG repeat expansion had a negative family history of HD. Of the 15 patients presenting in the first decade, 12 had greater than 80 CAG repeats and a clinical profile at the time of the test that included two or more of the following: declining school performance, seizures, oral motor dysfunction, rigidity, and gait disorder. Three patients with smaller CAG repeat expansions had incomplete or atypical symptom profiles. Symptom patterns in patients presenting in the second decade were more varied but usually included behavioral and motor symptoms. Patients without CAG expansions had incomplete or atypical symptom profiles. We define the historical and clinical profiles of HD presenting in the first two decades and suggest that physicians exercise restraint in using a "diagnostic" gene test for HD in the evaluation of at-risk children with incomplete or atypical symptom profiles or no family history of HD, in whom test results are very likely to be normal or unrelated to the patient's symptoms. PMID- 9339689 TI - Impaired inhibition in writer's cramp during voluntary muscle activation. AB - We used paired transcranial magnetic stimulation (TMS) to evaluate inhibitory mechanisms in eight patients with writer's cramp during rest and isometric wrist extension. Both stimuli were 110% of the motor threshold; the interstimulus intervals (ISIs) were 20 to 200 ms in increments of 10 ms. Surface EMG was recorded from wrist extensors. In the symptomatic hemisphere, there was no significant difference in the amplitude of the test (second) motor evoked potential (MEP) between patients and age-matched controls at rest. However, with voluntary muscle activation, inhibition of the test MEP by the conditioning MEP was significantly less in writer's cramp patients than in controls (p = 0.02). The difference was most prominent at ISIs of 60 to 80 ms in which inhibition is maximum. In the asymptomatic hemisphere, there was no significant difference between patients and controls in both rest and active conditions. The silent period was shorter in patients than controls on the symptomatic side (p = 0.003) but not on the asymptomatic side. We conclude that the inhibitory effects induced by magnetic stimulation are reduced in patients with writer's cramp, but only on the symptomatic side during muscle activation. This may relate to the overflow of muscle activity that characterizes this condition. PMID- 9339690 TI - Double-blind comparison of pramipexole and bromocriptine treatment with placebo in advanced Parkinson's disease. International Pramipexole-Bromocriptine Study Group. AB - Pramipexole is a new, selective, nonergoline dopamine agonist that acts on D2 and preferentially on D3 dopamine receptors. Phase II and III clinical trials have shown this drug to be useful in treating both early and advanced Parkinson's disease (PD) patients. A double-blind, randomized, multicenter study was performed to compare the safety, tolerance, and efficacy of pramipexole versus placebo in patients with advanced PD with motor fluctuations. A bromocriptine treatment group was included to enable comparisons between bromocriptine and placebo groups, but the study was not powered to show statistical differences between the active treatment groups. The study included 247 patients with "wearing off." Patients were Hoehn and Yahr stages II to IV during "on" times. The trial included three phases: dose escalation, 6 months' maintenance, and dose reduction. The primary end points were the Unified Parkinson's Disease Rating Scale (UPDRS) parts II and III. Up to 4.5 mg per day of pramipexole and 30 mg per day of bromocriptine were used. The results of the study showed that the UPDRS part II improved by 26.7% for pramipexole (p = 0.0002) and 14% for bromocriptine (p = 0.02) versus 4.8% for placebo. The UPDRS part III showed improvements of 34% for pramipexole (p = 0.0006) and 23.8% for bromocriptine (p = 0.01) versus 5.7% for placebo. There were no major differences in safety data. In the active treatment groups there were more reports of dyskinesia and nausea compared with placebo. In regard to comparison of the Global Clinical Assessment of Efficacy between active treatment groups, there was a trend to significance (p = 0.056) in favor of pramipexole. We conclude that pramipexole-treated patients with advanced PD improved significantly more than placebo for both primary end points. PMID- 9339691 TI - Tolcapone improves motor function in parkinsonian patients with the "wearing-off" phenomenon: a double-blind, placebo-controlled, multicenter trial. AB - We studied the new catechol-O-methyltransferase inhibitor tolcapone, 100 and 200 mg, three times daily (tid) in a randomized, double-blind, parallel-group trial involving 202 parkinsonian patients who were experiencing the "wearing-off" phenomenon on levodopa therapy. After 3 months, patients receiving tolcapone had a significant decrease in mean daily levodopa dose requirement compared with placebo-treated patients (p < 0.01). In patients treated with tolcapone 200 mg tid, daily "off" time, measured using patient diaries, was reduced from baseline by 3.25 hours; this reduction was significantly different from that seen in the placebo group (p < 0.01). Moreover, the number of daily levodopa intakes was reduced significantly in each tolcapone group compared with placebo (p < 0.01). We found significant improvements in motor function and overall efficacy in the tolcapone groups (p < 0.01). The most frequent adverse events were associated with levodopa treatment. Dyskinesia developed or worsened in 18% of patients receiving placebo, in 51% receiving tolcapone 100 mg tid, and in 64% receiving 200 mg tid, with most cases occurring within the first 30 days of the study. Diarrhea was the most frequent nondopaminergic event, occurring in 14% on placebo, 13% on tolcapone 100 mg tid, and 19% on 200 mg tid. Overall 18% of patients withdrew because of adverse events: 15% on placebo, 17% on tolcapone 100 mg tid, and 22% on 200 mg tid. We conclude that tolcapone as an adjunct offers promise for the relief of the "wearing-off" phenomenon in levodopa-treated parkinsonian patients. PMID- 9339692 TI - Unilateral pallidotomy for Parkinson's disease: comparison of outcome in younger versus elderly patients. AB - We studied the effects of medial pallidotomy in the first 20 consecutive patients with Parkinson's disease (PD) undergoing this MRI/electrophysiologically guided procedure at our institution. The mean age of patients was 65.5 years (median 66.5) and none suffered any serious complications. Pallidotomy significantly improved motor function in both "on" and "off" states as measured by Unified Parkinson's Disease Rating Scale (UPDRS) motor scores and timed tests (Purdue pegboard and counter tapping) in the arm contralateral to surgery 3 months postoperatively. Patients also improved in terms of activities of daily living, reflected by improved UPDRS activity of daily living and complications of therapy scoring and reduced levodopa-induced dyskinesias; six of 11 patients who could not walk in an "off" state prior to surgery could do so postoperatively. The total UPDRS score improved by 22% from preoperative values. The aforementioned improvements occurred similarly in patients greater than (n = 11) or less than 65 years (n = 9) at surgery. Neuropsychological measures indicated that although the majority of cognitive function remains unchanged in right-handed PD patients following dominant (left) hemisphere pallidotomy, mild specific declines in word generation are present. The findings of this study suggest that unilateral pallidotomy is safe and associated with improved motor functioning in elderly as well as younger PD patients experiencing significant disability despite optimal medical therapy. PMID- 9339693 TI - Unilateral pallidal stimulation for Parkinson's disease: neurobehavioral functioning before and 3 months after electrode implantation. AB - Unilateral pallidotomy is thought to have a low risk for cognitive morbidity. Nonetheless, recent research suggests that some patients experience declines in memory and language and that pallidal stimulation might be a safer treatment for Parkinson's disease (PD). We investigated the neurobehavioral effects of unilateral pallidal stimulation. Nine consecutive PD patients undergoing unilateral deep brain-stimulating electrode implantation in the globus pallidus interna were evaluated with a neuropsychological test battery approximately 1 month before and 3 months after surgery. Patients reported significantly fewer symptoms of anxiety and greater vigor after surgery. There was a trend toward fewer depressive symptoms. Semantic verbal fluency and visuoconstructional test scores declined significantly after surgery. However, among five patients showing declines in semantic verbal fluency, only one patient's score declined by more than 2 SD. No patient showed significant decline or improvement in the overall level of cognitive functioning. This study supports the relative safety, in terms of cognitive function, of unilateral pallidal stimulation in PD. PMID- 9339694 TI - Preoperative indicators of clinical outcome following stereotaxic pallidotomy. AB - We assessed the utility of preoperative clinical assessment and functional brain imaging with 18F-fluorodeoxyglucose (FDG) and positron emission tomography (PET) in predicting the clinical outcome of stereotaxic pallidotomy for the treatment of advanced Parkinson's disease (PD). Twenty-two PD patients undergoing posteroventral pallidotomy were assessed preoperatively with the Core Assessment Program for Intracerebral Transplantation (CAPIT) ratings measured on and off levodopa; quantitative FDG/PET was also performed before surgery. Preoperative clinical and metabolic measurements were correlated with changes in off-state CAPIT ratings determined 3 months after surgery. Clinical outcome following pallidotomy was also correlated with intraoperative measures of spontaneous pallidal single-unit activity as well as postoperative MRI measurements of lesion volume and location. We found that unilateral pallidotomy resulted in variable clinical improvement in off-state CAPIT scores for the contralateral limbs (mean change 30.9 +/- 15.5%). Postoperative MRI revealed that pallidotomy lesions were comparable in location and volume across the patients. Clinical outcome following surgery correlated significantly with preoperative measures of CAPIT score change with levodopa administration (r = 0.60, p < 0.005) and with preoperative FDG/PET measurements of lentiform glucose metabolism (r = 0.71, p < 0.0005). Operative outcome did not correlate with intraoperative measures of spontaneous pallidal neuronal firing rate. We conclude that preoperative measurements of lentiform glucose metabolism and levodopa responsiveness may be useful indicators of motor improvement following pallidotomy. Both preoperative quantitative measures, either singly or in combination, may be helpful in selecting optimal candidates for surgery. PMID- 9339695 TI - Mass effect and death from severe acute stroke. AB - BACKGROUND: In severe acute stroke, the degree of midline cerebral displacement is related to level of consciousness but not to survival. Early identification of patients at high risk of death from mass effect would assist patient management decisions. METHODS: We measured lesion volume, horizontal pineal displacement (PD), and horizontal septum pellucidum displacement (SD) on axial CT of consecutive patients with severe (Canadian Neurological Scale score < or = 5) acute hemispheric stroke. We correlated CT measurements with the probability of 14-day survival. RESULTS: Forty-six (39%) of 118 patients died within 14 days and 72 (61%) died within 1 year following stroke. Crude risk factors for 14-day mortality were as follows: lesion volume > or = 400 ml, SD > or = 9 mm, PD > or = 4 mm, intraventricular hemorrhage, and coma on admission. Only SD (p = 0.001) and coma on admission (p = 0.019) remained significant in multivariate analysis, but PD was highly correlated with SD (r = 0.82). PD of > or = 4 mm on a scan performed within 48 hours of stroke onset identified patients with a low probability of 14-day survival (0.16; CI 0 to 0.32) with a specificity of 89% and a sensitivity of 46%. CONCLUSIONS: The degree of horizontal midline cerebral displacement correlates with the likelihood of death following stroke. Patients with > or = 4 mm PD on CT performed within 48 hours of stroke onset are at high risk for early death. PMID- 9339696 TI - Meta-analysis of the Hachinski Ischemic Score in pathologically verified dementias. AB - Our objectives were to investigate the utility of the Hachinski Ischemic Score (HIS) in differentiating patients with pathologically verified Alzheimer's disease (AD), multi-infarct dementia (MID), and "mixed" (AD plus cerebrovascular disease) dementia, and to identify the specific items of the HIS that best discriminate those dementia subtypes. Investigators from six sites participated in a meta-analysis by contributing original clinical data, HIS, and pathologic diagnoses on 312 patients with dementia (AD, 191; MID, 80; and mixed, 41). Sensitivity and specificity of the HIS were calculated based on varied cutoffs using receiver-operator characteristic curves. Logistic regression analyses were performed to compare each pair of diagnostic groups to obtain the odds ratio (OR) for each HIS item. The mean HIS (+/- SD) was 5.4 +/- 4.5 and differed significantly among the groups (AD, 3.1 +/- 2.5; MID, 10.5 +/- 4.1; mixed, 7.7 +/ 4.3). Receiver-operator characteristic curves showed that the best cutoff was < or = 4 for AD and > or = 7 for MID, as originally proposed, with a sensitivity of 89.0% and a specificity of 89.3%. For the comparison of MID versus mixed the sensitivity was 93.1% and the specificity was 17.2%, whereas for AD versus mixed the sensitivity was 83.8% and the specificity was 29.4%. HIS items distinguishing MID from AD were stepwise deterioration (OR, 6.06), fluctuating course (OR, 7.60), hypertension (OR, 4.30), history of stroke (OR, 4.30), and focal neurologic symptoms (OR, 4.40). Only stepwise deterioration (OR, 3.97) and emotional incontinence (OR, 3.39) distinguished MID from mixed, and only fluctuating course (OR, 0.20) and history of stroke (OR, 0.08) distinguished AD from mixed. Our findings suggest that the HIS performed well in the differentiation between AD and MID, the purpose for which it was originally designed, but that the clinical diagnosis of mixed dementia remains difficult. Further prospective studies of the HIS should include additional clinical and neuroimaging variables to permit objective refinement of the scale and improve its ability to identify patients with mixed dementia. PMID- 9339697 TI - Lesions in the left arcuate fasciculus region and depressive symptoms in multiple sclerosis. AB - Depression is a common mood disturbance in multiple sclerosis (MS) patients. Epidemiologic data suggest a causative relationship between depressive symptoms and cerebral demyelination, although a specific lesion site responsible for depressed mood has not been identified. Given that depression in neurologic disease is closely related to frontal and temporal lobe damage, we focused our study on investigating the extent to which lesions in the white matter connecting both cerebral lobes may account for depressive symptoms in MS. Forty-five patients were assessed using the Beck Depression Inventory and an MRI protocol conceived to quantify lesions separately in the basal, medial, and lateral frontotemporal white matter. The presence of lesions in the left suprainsular white matter, the region that mainly includes the arcuate fasciculus, was specifically associated with depressive symptoms, accounting for a significant 17% of the depression score variance. Although a multifactorial origin is suspected for depression in MS, this finding gives support to the existence of a direct negative effect of demyelination on mood. PMID- 9339698 TI - VLA-4 expression on peripheral blood lymphocytes is downregulated after treatment of multiple sclerosis with interferon beta. AB - Adhesion molecules are likely to play a critical role in the immunopathogenesis of multiple sclerosis (MS). The interaction of vascular cell adhesion molecule-1 (VCAM-1) with its lymphocyte ligand very late antigen-4 (VLA-4) may mediate migration of lymphocytes into the CNS. We have previously demonstrated that MS patients treated with interferon beta (IFN-beta) have a significant increase in soluble VCAM-1 (sVCAM-1) soon after the initiation of treatment, and this effect correlated with the resolution of contrast-enhancing MRI lesions. We studied the cell surface expression of VLA-4 by flow cytometry in 10 MS patients before and during IFN-beta treatment. We found a significant decrease in mean VLA-4 fluorescence of MS patients' lymphocytes on treatment and no change in untreated controls. In vitro treatment of lymphocytes with IFN-beta did not reproduce this effect, but the addition of sVCAM-1 did result in a decrease in VLA-4 expression. These data indicate that the previously identified increase in sVCAM-1 may lead to a decrease in VLA-4 expression and that this effect may partially explain the mechanism of action of IFN-beta. PMID- 9339700 TI - Assessment: dermatomal somatosensory evoked potentials. Report of the American Academy of Neurology's Therapeutics and Technology Assessments Subcommittee. PMID- 9339699 TI - Human T-cell response to myelin basic protein peptide (83-99): extensive heterogeneity in antigen recognition, function, and phenotype. AB - Multiple sclerosis (MS) is considered a T cell-mediated autoimmune disease, and myelin proteins are the most likely candidate autoantigens. Based on experiments performed in experimental allergic encephalomyelitis (EAE), innovative immunotherapies have been developed that target either the specific trimolecular complex of encephalitogenic T cells, consisting of T-cell receptor (TCR), major histocompatibility complex (MHC; HLA in humans) class II molecule, and autoantigenic peptide, or the effector functions of these cells. To provide the basis for the transfer of these specific immunotherapies to MS, we extensively characterized the human T-cell response to one major myelin epitope, the myelin basic protein peptide (83-99). We analyzed restriction element, TCR usage and affinity, fine specificity, cytokine production, cytolytic activity, and expression of surface molecules on 41 T-cell clones (TCCs) derived from MS patients and normal controls. We demonstrate a high degree of complexity of recognition patterns as well as of functional phenotypes among T cells responding to the same epitope. In contrast to results from animal models, these findings indicate that the design of epitope-based specific immunotherapies for MS is more difficult than previously thought. PMID- 9339701 TI - CSF antigliadin antibodies and the Ramsay Hunt syndrome. AB - Although the association between celiac disease and progressive myoclonic ataxia is well recognized, in each of the reported cases the neurologic features began in middle adult life and usually in patients who had clinical or laboratory evidence of malabsorption. We report a case of progressive myoclonic ataxia and epilepsy (Ramsay Hunt syndrome) that began in childhood. In this patient there were no features suggestive of gluten intolerance. The presence of antigliadin antibodies in the serum and CSF suggested celiac disease was the cause of the patient's neurologic syndrome. Duodenal morphologic abnormalities reversed with treatment but no major changes were noted in the patient. Celiac disease should be considered in the differential diagnosis of myoclonic ataxia at any age, even in the absence of clinical evidence of gluten-sensitive enteropathy. PMID- 9339702 TI - Fluorodopa and raclopride PET analysis of patients with Machado-Joseph disease. AB - We performed [18F]6-fluoro-L-dopa (6-FD) and [11C]raclopride (RAC) PET studies in six patients with Machado-Joseph disease (MJD) (age, 17 to 61 years; duration of illness, 3 to 10 years), normal controls (n = 10 in 6-FD-PET, n = 8 in RAC-PET), and patients with idiopathic parkinsonism (n = 15 in 6-FD-PET). The youngest patient with MJD had prominent dystonia and pyramidal features (type 1 MJD), whereas the remainder were prominently ataxic (types 2 and 3 MJD). Striatal RAC binding was normal in patients with MJD. Striatal 6-FD influx constants (Ki) were low in the range of idiopathic parkinsonism in two patients with MJD (youngest and oldest patients), whereas striatal Ki were normal in the remaining patients with MJD. The impairment of the nigrostriatal dopaminergic pathway did not correlate with the phenotype, CAG repeat length, disease duration, or age of onset of patients with MJD. Our results suggest that striatal D2 receptors are normal and the nigral damage is diverse in MJD. PMID- 9339703 TI - De novo 14484 mitochondrial DNA mutation in monozygotic twins discordant for Leber's hereditary optic neuropathy. AB - Monozygotic twin brothers, clinically discordant for Leber's hereditary optic neuropathy (LHON), had a heteroplasmic point mutation at position 14484 in the mitochondrial DNA that was not detected in their mother. Moreover, the mutation occurred on the rare European haplogroup X, rather than the haplogroup J commonly associated with the 14484 mutation. These data indicate that the 14484 mutation in this family was a new mutation, indicating that it was the de novo occurrence of a common, primary LHON mutation. PMID- 9339705 TI - Trigeminal neuralgia in patients with multiple sclerosis: lesion localization with magnetic resonance imaging. AB - We performed conventional T2-weighted brain MRI examinations in six patients with multiple sclerosis (MS) and trigeminal neuralgia. In all patients brainstem lesions in positions expected to involve trigeminal fibers, particularly the entry zone of sensory fibers, were demonstrated. Compression of the trigeminal nerve by ectatic vessels, a recognized cause of idiopathic trigeminal neuralgia, was not observed. We conclude that in MS trigeminal neuralgia is usually caused by demyelinating lesions affecting pontine trigeminal pathways. PMID- 9339704 TI - Axonal dysfunction and disability in a relapse of multiple sclerosis: longitudinal study of a patient. AB - In a 6-year longitudinal study of a patient with relapsing progressive multiple sclerosis (MS), we used proton magnetic resonance spectroscopy to assess N acetylaspartate (NAA) from a large central brain volume to evaluate the relationship between this marker of neuronal integrity and clinical disability. During the follow-up period, there was one major relapse and its subsequent partial remission. Changes in the brain NAA to creatine ratio correlated strongly with clinical disability (Spearman rank coefficient = -0.73, p < 0.001). We interpret this as evidence that axonal dysfunction or loss contributes to functional impairment of patients with MS. Because the NAA signal in the large volume of interest originated predominantly from white matter that appeared normal on conventional MRI, these results also suggest that some degree of axonal dysfunction may be widespread in acute, severe relapses. PMID- 9339706 TI - Early high-dose intravenous methylprednisolone in acute disseminated encephalomyelitis: a successful recovery. AB - We describe a patient with acute disseminated encephalomyelitis (ADEM) who was treated with high-dose intravenous methylprednisolone 2 days after onset of neurologic symptoms. Despite poor prognostic factors and extensive white matter lesions, the patient recovered dramatically with no need for maintenance steroid therapy. This case report of fulminant ADEM treated successfully with early high dose intravenous methylprednisolone, although uncontrolled, suggests that this agent should be studied in other cases. PMID- 9339707 TI - Focal transmantle dysplasia: a specific malformation of cortical development. AB - We report 18 patients who presented prior to age 20 years with epilepsy or fixed neurologic deficits. MRI showed signal abnormality extending from the cortex to the superolateral wall of the lateral ventricle. Histology showed cortical disorganization, neuronal cytomegaly, balloon cells, indistinct cortical gray matter-white matter junctions, and variable accompanying astrogliosis. We propose that this transmantle dysplasia is a specific anomaly resulting from abnormal stem cell development. PMID- 9339708 TI - Very late onset Friedreich's ataxia without cardiomyopathy is associated with limited GAA expansion in the X25 gene. AB - Molecular analysis of spinocerebellar ataxias revealed a pathologic GAA expansion in the gene encoding frataxin in six adult patients from three families. These patients, carrying expanded alleles in the low-range size, had an exceptionally late onset and lacked cardiomyopathy, pointing to phenotypic variability of Friedreich's ataxia. Both mitotic and gametic instability of the expanded triplet repeat were present in these families. PMID- 9339709 TI - Ganglionitis in paraneoplastic subacute sensory neuronopathy: a morphologic study. AB - A 69-year-old woman presented with subacute sensory neuropathy and autonomic dysfunction of 9 months' duration, associated with high serum titers of anti-Hu antibodies. A small cell carcinoma of the lung was diagnosed by biopsy. She died after cardiorespiratory arrest. At autopsy, spinal and autonomic ganglia showed subacute inflammation with diffuse endoneurial T-cell, B-cell, and plasma cell infiltration. The cytoplasm and nuclei of some ganglion neurons displayed IgG immunocytochemical positivity. CD8+ T cells were tightly attached to, and indented the cell surface of, IgG-positive and IgG-negative neurons. This observation suggests that both cytotoxic T-cell-mediated attack against neurons and humoral mechanisms play a role in paraneoplastic subacute sensory neuronopathy. PMID- 9339710 TI - Surgical treatment of carpal tunnel syndrome in patients with peripheral neuropathy. AB - Outcome after carpal tunnel surgery was studied retrospectively in 32 patients with peripheral neuropathy and carpal tunnel syndrome. Nocturnal paresthesias were almost universally relieved, followed in order of responsiveness by pain, numbness, and weakness. Twenty-five of 28 patients said they would have the surgery again if the outcome were the same. Patients with carpal tunnel syndrome and peripheral neuropathy benefit from surgical treatment of carpal tunnel syndrome. PMID- 9339711 TI - Clinical and molecular analysis of a pedigree of southern Italian ancestry with spinocerebellar ataxia type 2. AB - We describe patients from five generations of a pedigree with mutations in the spinocerebellar ataxia type 2 gene (SCA2). The predominant clinical features observed included both appendicular and truncal ataxia, dysarthria, slowness of saccades, and impaired optokinetic responses. Successive generations demonstrated both earlier ages of onset as well as increasing numbers of trinucleotide repeat sequences. The signs found in this family are compared with the description of other families with SCA2 as well as with other types of dominantly inherited spinocerebellar ataxias. PMID- 9339712 TI - A novel mitochondrial tRNA isoleucine gene mutation causing chronic progressive external ophthalmoplegia. AB - We report a sporadic case of chronic progressive external ophthalmoplegia that developed during childhood and was associated with ragged-red and cytochrome c oxidase (COX)-negative fibers in skeletal muscle. Sequencing of all the mitochondrial transfer RNA (tRNA) genes identified a single potentially pathogenic mutation--a T to C transition at position 4274 in the tRNA(Ile) gene. This mutation was not present in skeletal muscle from 79 controls, and the level of the mutation in COX-negative fibers was significantly greater than the level in COX-positive fibers. PMID- 9339714 TI - Shakespeare and sleep disorders. AB - Shakespeare was a consummate dramatist and profound observer of human behavior. He vividly described many clinical disorders, including those of sleep. His characters suffered from somnambulism, sleep apnea, insomnia, and nightmares. Sleep, to Shakespeare, was a blessing denied to many of his protagonists. PMID- 9339715 TI - Stereotactic selective Vo-complex thalamotomy in a patient with dystonic writer's cramp. PMID- 9339713 TI - A survey of antidepressant drug use in Parkinson's disease. Parkinson Study Group. AB - Selective serotonin reuptake inhibitors (SSRIs) are a newer class of antidepressants that may have particular efficacy in Parkinson's disease (PD) given the known serotonergic alterations in this disease. These agents are also thought to have a favorable side-effect profile, particularly in the elderly. Several recent case reports, however, have raised concern that SSRIs may worsen parkinsonian motor function. We surveyed 71 Parkinson Study Group (PSG) investigators using a standardized questionnaire about their usage of antidepressants in PD. Based on estimates provided by 49 investigators (70%) (caring for approximately 23,410 PD patients) who responded, 26% of patients with PD are on pharmacotherapy for depression. These physicians use SSRIs as first line therapy 51% of the time, tricyclic antidepressants 41% of the time and other agents 8% of the time. The most common reasons for selecting SSRIs were their better side-effect profile and perceived greater efficacy. The most common reasons for selecting tricyclic antidepressants were their potential to help with sleep and the physician's experience with this class. Forty-three percent of investigators were concerned that SSRIs might worsen motor function, and 37% of them have had at least one patient in whom they believe this had occurred. Our survey confirms that for treating physicians there remain uncertainties regarding the relative efficacy and tolerability of available antidepressant medications for patients with PD. A controlled clinical trial of antidepressant therapy in PD would be valuable for settling these concerns. PMID- 9339716 TI - Post-transfusion reversible posterior leukoencephalopathy syndrome with cerebral vasoconstriction. PMID- 9339717 TI - Microscope field size and the neuropathologic criteria for Alzheimer's disease. PMID- 9339718 TI - Brachial plexus neuropathy after botulinum toxin injection. PMID- 9339719 TI - Assessment of neuropsychological testing. PMID- 9339720 TI - Assessment of neuropsychological testing. PMID- 9339721 TI - Assessment of neuropsychological testing. PMID- 9339722 TI - Assessment of neuropsychological testing. PMID- 9339723 TI - Assessment of neuropsychological testing. PMID- 9339724 TI - Assessment of neuropsychological testing. PMID- 9339725 TI - Assessment of neuropsychological testing. PMID- 9339726 TI - Assessment of neuropsychological testing. PMID- 9339727 TI - Assessment of neuropsychological testing. PMID- 9339728 TI - Assessment of neuropsychological testing. PMID- 9339729 TI - International submissions to Neurology. PMID- 9339730 TI - Migraine-associated hypotension and autonomic ganglionitis. PMID- 9339731 TI - Long-term prognosis of typical childhood absence epilepsy. PMID- 9339732 TI - Markers of reversible hepatic encephalopathy. PMID- 9339733 TI - Transient encephalopathy after Taxol infusion. PMID- 9339734 TI - Sleep apnea syndrome among poliomyelitis survivors. PMID- 9339735 TI - Red ear syndrome. PMID- 9339736 TI - Chronic progressive monomelic sensory neuropathy. PMID- 9339737 TI - Myelopathy and coinfection with HIV and HTLV-I. PMID- 9339738 TI - Immune alterations in geriatric mice dosed subacutely with diethylstilbestrol (DES). AB - Exposure to both physiological and pharmacological doses of estrogenic compounds has been reported to alter immunologic responses in humans as well as in developmental and adult murine models. Despite the current therapeutic use of potent estrogens, including diethylstilbestrol (DES), in geriatric human disorders, elucidation of the effects of these agents on the aged immune system is limited. The present report describes highly significant alterations in the thymus and bone marrow of aged mice (21 +/- 1 months) exposed subacutely to DES. Severe thymic hypocellularity developed in treated mice following five consecutive days of intraperitoneal injection with 1.5 or 6.0 mg kg(-1) DES. Cell maturation within the thymus was also affected, as indicated by a significant decrease in CD4+8+ cells and a concomitant increase in CD4-8- cells. Cellularity of the bone marow was unchanged by DES. However, significant changes were observed in percentages of bone marrow cells expressing surface antigens CD45 (common leukocyte lineage), Mac-1 (macrophage lineage) and CD45R (B-lineage lymphocytes). Both percentages and total numbers of cells in the spleen expressing Thy 1.2 (T-lineage lymphocytes) were also reduced. These immune changes in geriatric mice exposed to DES were similar in direction but more severe than those reported in either young adult or perinatal models. These data may suggest a need for considering the geriatric immune system separately from other age groups when determining effects of immunosuppressive or immunotoxic compound exposure. PMID- 9339739 TI - Effects of a new phosphorothionate (RPR-V) on ATPases and acetylcholinesterase in rat brain by subchronic dosing. AB - The effect of a new phosphorothionate, the ethyl ester of 2-butenoic acid 3 diethoxy phosphinothioyl (RPR-V) synthesized at the Indian Institute of Chemical Technology, Hyderabad was studied using peroral doses of 0.033, 0.066 and 0.099 mg kg(-1) in male and female rats daily over 90 days. This repeated administration of RPR-V caused significant inhibition of acetylcholinesterase, Na+-K+, Mg2+ and Ca2+-ATPases in the brain of male and female rats when measured after 45 and 90 days of treatment. The effects of the low dose were generally not statistically significant, whereas medium and high doses caused significant effects. Females were more susceptible with regard to brain AChE, but the reverse was seen with Mg2+-ATPase, suggesting sexual dimorphism. Enzyme recoveries were seen 28 days after the final dose. Since RPR-V not only inhibited AChE but also ATPases, it is possible that both synaptic transmission and nerve conduction were affected. PMID- 9339740 TI - Alterations in the cardiopulmonary effects and pharmacokinetics of a bisphosphonate drug by a cytochrome P-450 inhibitor in conscious rats. AB - U-91502, a bisphosphonate for arthritic inflammation treatment, was evaluated for its parental toxicity. The objective was to differentiate between the parent drug and a reactive metabolite(s) as the proximate cause of the toxic effects using two methods. The first method was to block the metabolism of U-91502 with a broad spectrum cytochrome P-450 inhibitor, 1-aminobenzotriazole (ABT), to increase its toxicity. The second method was to scavenge any electrophilic intermediates of U 91502 with supplemental nucleophiles, L-methionine (LM) and N-acetylcysteine (NaLc) to decrease its toxicity. Two groups of rats each were given an i.v. injection of saline or ABT followed by an i.v. infusion of U-91502 at a constant dose rate. A third group was given two oral doses of LM followed by a co-infusion of U-91502 and NaLc. The breathing rate (BR) and electrocardiogram (ECG) of the rats were monitored. Blood samples were taken at specified time points for plasma drug concentration analyses (PDC) and pharmacokinetics determination. Each rat was infused until its BR was depressed by approximately 30% from the rates prior to injection of saline or ABT, or the second oral dose of LM. Thereafter, half of the rats in each group were sacrificed immediately and the remaining half at 180 min post infusion. All infused rats, except for those of the co-infusion group, and a group of untreated rats were analyzed for hepatic non-protein sulfhydryl for indication of glutathione depletion. The results indicated that ABT pretreatment expedited the elevation of PDC to a critical level that caused BR and then heart rate (HR) depression and ECG alterations. There was no unusual depletion of glutathione. The maximum concentration and the area under the curve were significantly increased while the total clearance was significantly reduced. Consequently, the postinfusion PDC remained high and the BR and HR depressions persisted. LM and NaLc did not alleviate the toxicity or alter the pharmacokinetics of U-91502. It was concluded that the toxic effects of U-91502 were due mainly to the parent drug and not the metabolites. PMID- 9339741 TI - Vitamin C supplementation on hepatic oxidative stress induced by cigarette smoke. AB - A study has been conducted to investigate whether the oxidative damage produced in the liver of rats exposed to cigarette smoke can be effectively combatted with vitamin C, one of the antioxidant vitamins. We assessed the liver antioxidants (vitamins E, C and A), scavenging enzymes and lipid peroxide products of rats exposed to cigarette smoke and simultaneously given vitamin C (200 mg 100 g[-1] body wt.) for 90 days. Malondialdehyde (MDA), conjugated dienes, hydroperoxides and free fatty acids (FFA) were significantly increased in liver of smoke-exposed groups. The activity of superoxide dismutase and catalase and vitamin E and C contents were significantly lower than controls. But vitamin A, glutathione (GSH) content and glutathione peroxidase (GSH Pxase) activity were enhanced. Vitamin C supplementation to smoke-exposed rats showed increased resistance to lipid peroxidation and increased activity of scavenging enzymes. The GSH content, vitamin C and FFA were brought to normal levels. Thus, this study seems to suggest that an intake of a mega dose of vitamin C can protect the liver from oxidant damage caused by cigarette smoke. PMID- 9339742 TI - Histology of ovaries and uteri and levels of plasma progesterone, oestradiol 17beta and oestrone sulphate during the implantation period in mated and gonadotrophin-releasing hormone-treated mink (Mustela vison) exposed to polychlorinated biphenyls. AB - Earlier studies have shown that polychlorinated biphenyls (PCB) do not prevent ovulation, fertilization and implantation, but exposure of female mink during gestation caused fetal death. To understand this phenomenon, 30 PCB-exposed female mink and 30 controls were mated or induced to ovulate without fertilization by treatment with gonadotrophin-releasing hormone (GnRH) and correlations were measured between reproductive, morphological and endocrine parameters. The exposure to PCB (Aroclor 1254) started before ovulation. Equal numbers of animals from each group were killed on days 10, 17 and 26 post-coitum or on GnRH-injection days. Plasma concentrations of progesterone, oestradiol 17beta and oestrone sulphate in sampled blood were then measured. Ovaries, uteri and placentae were examined histologically. Compared with controls, exposure to PCB during the reproductive season resulted in significantly smaller uterine glandular diameters before implantation or at the end of the experiment in both mated and GnRH-treated mink. The GnRH treatment increased the sizes of the ovarian corpora lutea and oesterone sulphate 10 days after treatment but the increase in uterine glandular diameter was significant only in GnRH-treated control animals. Both GnRH-treated and mated PCB-exposed groups displayed a peak in oesterone sulphate concentration that was not observed in any of the control groups. Possible actions of PCB are discussed. The observed histological and hormonal changes in the mated PCB-exposed group did not prevent implantation. Exposure to PCB increased fetal mortality. This effect was associated with an effect on placental development. PMID- 9339743 TI - Chronic ethanol intake induces oxidative alterations in rat testis. AB - Although it is well known that chronic ethanol abuse produces sexual dysfunction and impaired spermatogenesis, the mechanisms of ethanol-induced testicular alterations are not fully explained. Therefore, the aim of this study was to investigate the mechanisms of testicular oxidative damage in rats given drinking water containing 3% ethanol for 8 weeks. Control rats were pair-fed with saccharose. The mean daily ethanol intake was 4.05 g kg(-1), corresponding to the consumption of 41 of wine (10% alcohol) or 0.71 of whiskey (40% alcohol) by a man of 70 kg body wt. Exposure to ethanol caused a significant depletion in the testicular levels of glutathione (GSH), protein containing sulfhydryl groups, tocopherol and ascorbic acid, and an increase in the concentrations of malondialdehyde (index of lipid peroxidation) and carbonyl proteins (index of protein oxidation). Other effects were decreases in the concentration of adenosine 5'-triphosphate and in the activity of glutathione peroxidase, and an increase in the activity of alcohol dehydrogenase. In summary, this study shows that in the rat, daily consumption of ethanol in the drinking water increases lipid and protein oxidation. In addition to impaired antioxidant defence, an imbalance in energy production may also play a role in the toxic reaction to alcohol. PMID- 9339744 TI - Miconazole genotoxicity in mice. AB - The genotoxic effects of miconazole (MC) were studied in mouse bone-marrow cells and primary spermatocytes at diakinesis metaphase I of meiosis. The ability of miconazole to induce chromosomal aberrations was investigated. Both acute and subacute treatments were tested. Doses were 0.1, 0.5 and 1 mg per animal. Both acute and subacute treatments induced statistically significant dose-dependent chromosomal aberrations. The effect of miconazole on sperm head morphology was also studied in animals treated for five successive days with the three doses. Morphological sperm head abnormalities increased significantly after treatment with miconazole. The increase was dose-dependent. These results suggests that miconazole has a genotoxic effect on mice somatic and germ cells. PMID- 9339745 TI - Effect of cadmium on morphology and steroidogenesis of cultured human ovarian granulosa cells. AB - Cadmium (Cd) is able to decrease preovulatory luteinizing hormone (LH) levels in blood and inhibit ovulation in rats. In this study the direct effects of Cd on steroidogenesis in granulosa cells were investigated. The cells obtained from ovarian follicular aspirates of 41 women undergoing in vitro fertilization (IVF) were cultured. Cadmium-induced alterations in the cellular morphology and in the production of progesterone by the cells was determined after exposure to concentrations of 8, 16, 32 and 64 microM CdCl2 for 2, 4, 8, 24 and 48 h. Progesterone secretion by granulosa cells could be stimulated with increasing concentrations of follicle-stimulating hormone (FSH). Combined effects of Cd and FSH were also studied. Cadmium diminished progesterone production in unstimulated and FSH-supported cells depending on its concentration and the exposure time. Follicle-stimulating hormone (100 ng ml[-1]) protected against Cd-induced suppression of progesterone production. Cadmium interfered with cell-cell junctions and the adherence of cells. No protective effect of FSH on Cd-induced alteration in cell morphology could be observed. Retraction of cytoplasmic extensions occurred at a lower dose and within a shorter exposure than a decrease in progesterone production. In conclusion, Cd exerted a direct effect on both granulosa cell morphology and on steroid biosynthesis. The lowest Cd concentration (16 microM) that was able to reduce progesterone production was about 3.5 times higher than levels reported in the ovary of a female smoker. The presented data can help to define environmental, occupational and life-style (smoking) risk factors in gonadal function during the preconception period of the female reproductive lifespan. PMID- 9339746 TI - Interaction of exercise training and chronic ethanol ingestion on antioxidant system of rat brain regions. AB - This study investigates the interactive effects of chronic ethanol ingestion and exercise training on the antioxidant system and lipid peroxidation in cortex, cerebellum, medulla, striatum and hypothalamus of the rat brain. Exercise training (6.5 weeks) significantly increased superoxide dismutase (SOD) activity in striatum, the region associated with motor activity, but decreased SOD activity in other brain regions. Catalase (CAT) activity decreased significantly in hypothalamus, the region associated with behavior, due to exercise. The training significantly increased glutathione peroxidase (GSH-Px) activity in brain regions studied with the exception of cerebellum. In addition, glutathione reductase (GR) activity increased in brain regions, with the exception of medulla. The training significantly decreased malondialdehyde (MDA) levels in all brain regions studied, which is due to training adaptation. Ethanol (20%) (2.0 g kg[-1], p.o. for 6.5 weeks) significantly decreased SOD activity in all regions except cortex, CAT activity in cortex, striatum and hypothalamus, GSH-Px activity in cerebellum and GR activity in medulla. Similarly, ethanol significantly decreased the GSH level in cortex, medulla and striatum and the GSH/GSSG ratio in medulla and cerebellum. Conversely, ethanol significantly augmented GR activity in cortex, cerebellum and striatum. When ethanol and exercise were combined, there was significantly increased SOD and CAT activity in striatum, GSH-Px activity in cortex, striatum and hypothalamus and GR activity in cortex and striatum. The GSH level was significantly depleted in cortex, striatum and medulla. Combining training and ethanol also decreased MDA levels in medulla and cerebellum. In conclusion, the sensitivity of specific brain regions in reaction to chronic ethanol ingestion or training is a function of variability in antioxidant system activity. Thus, exercise training protects specific brain regions against ethanol-induced oxidative injury. PMID- 9339747 TI - Cadmium and selenium in blood and urine related to smoking habits and previous exposure to mercury vapour. AB - The object of this work was to investigate possible interactions of mercury, cadmium and selenium in humans. Selenium and cadmium in blood and urine were determined in this cross-sectional study of 130 males, of whom 77 had been previously exposed to mercury vapour at a chloralkali plant. Of the participants, 61.5% were smokers and 16.2% were never-smokers. The concentration of selenium in blood (B-Se) was significantly lower in subjects currently smoking more than 50 g of tobacco per week compared to never-smokers, whereas the concentration of cadmium in blood (B-Cd) was significantly higher in all categories of current smokers. In the multiple linear regression analysis, B-Se as a dependent variable was negatively associated with B-Cd, whereas current smoking habits were not included in the model as a predictor variable. In contrast, B-Cd as a dependent variable was positively associated with current as well as previous smoking habits, and negatively with both B-Se and the 'cumulative dose' of previous mercury vapour exposure. The concentration of selenium in blood was also negatively associated with B-Cd in the group of never-smokers (Spearman's r = 0.80; P < 0.001). In conclusion, these results suggest a depressive effect of cadmium on the concentration of selenium in blood, while smoking alone did not operate as a true predictor for this effect. Furthermore, previous exposure to mercury apparently modifies the concentration of cadmium in blood. PMID- 9339748 TI - An improved limited sampling method for individualised busulphan dosing in bone marrow transplantation in children. AB - Busulphan (BU) pharmacokinetic (PK) studies in children undergoing bone marrow transplantation suggest that individual BU dosing may be necessary to optimise BU systemic exposure. Optimising BU systemic exposure may improve outcome and decrease toxicity in BMT. Because of practical limitations in obtaining blood from children and for financial reasons, a limited sampling method (LSM) is needed. However, such methods for BU have not been validated in children. In the present study, we individualized oral BU dosing in 10 children to target an area under the curve of BU (BU AUC) of 900-1400 microM/min based on BU AUC(0-infinity) calculated from nine serum BU concentrations performed after a BU test dose of 40 mg/m2. We validated a LSM using 3 BU concentrations to determine AUC. Six of nine patients studied (one patient non-evaluable), required their doses modified (3, lower; 3, higher). The mean percent dose change was 26.2% (range -33.3% to +45.3%). Our three sample LSM BU AUC(0-infinity) (1098 +/- 344, mean +/- 1 s.d.) correlated highly with our nine sample BU AUC(0-infinity) (1132 +/- 389, Pearson r = 0.98, P = 0.0001) and was not significantly different by t-test (P = 0.3). The mean percentage difference between the three sample LSM AUCs and the nine sample AUCs in each of our patients was 7.5%, (range -10.99% to +9.4%). Trough levels correlated extremely well with AUC (r = 0.95, P = 0.0001). Individual BU dosing, based on AUC, is necessary in most children to achieve targeted levels of BU therapy. An LSM of three BU concentrations performed at 0.5 h, 1 h and 6 h post-BU test dose closely predicts the AUC calculated from nine sampling points. PMID- 9339749 TI - Inhibition of lymphocyte blastogenic response in healthy donors treated with recombinant human granulocyte colony-stimulating factor (rhG-CSF): possible role of lactoferrin and interleukin-1 receptor antagonist. AB - The effects of rhG-CSF on lymphocyte blastogenesis were evaluated in six healthy donors, submitted to progenitor cell mobilization for allogeneic transplantation. Neutrophil, monocyte and lymphocyte count increased 6.7-fold, 5.3-fold and 2.0 fold on day +4 of rhG-CSF as compared with baseline. The DNA stimulation index (DNA SI) of 72 h phytohemagglutinin (PHA)-treated cultures decreased from 20% (15 35.5) prior to rhG-CSF to 6.7% (1.5-11.9; P = 0.0026), 8% (4-12; P = 0.0091) and 15% (9-22; P = 0.0091) on days +2, +4 and +6; similarly, reactivity to concanavalin A decreased from 18% (12-20) to 1.8% (0.5-7; P < 0.01), 3% (2-8; P < 0.01) and 5% (2-11; P = 0.009). No changes of lymphocyte response to pokeweed mitogen were observed. DNA SI of PHA-treated cultures inversely correlated with neutrophil and monocyte count. IL-1 receptor antagonist (IL-1ra) and lactoferrin (Lf) plasma levels sharply increased and correlated with neutrophil and monocyte count. IL-10 increased five-fold on day +2, returned to pretreatment values thereafter and did not show any correlation with DNA SI, suggesting that it was not responsible for the observed phenomena. Interestingly, DNA SI of PHA-treated cultures inversely correlated with IL-1ra and Lf levels. CD3+ and CD19+ lymphocyte activation status, ie CD23, CD25, CD30 and HLA-DR coexpression, was not affected by rhG-CSF administration. Pharmacological doses of rhG-CSF in healthy donors inhibit lymphocyte blastogenesis via an increased production and/or release of immunoregulatory soluble mediators, ie IL-1ra and Lf, by primed neutrophils and monocytes. PMID- 9339750 TI - High interleukin-10 serum levels are associated with fatal outcome in patients after bone marrow transplantation. AB - IL-10 plays an important role in the control of immune reactions during systemic infection. Here, IL-10 serum levels were investigated in patients after BMT. The IL-10 levels correlated with the clinical course of the patients and with serum levels of C-reactive protein (CRP) and neopterin (NP). A total of 26 patients with AML (7), ALL (12), CML (2), NHL (3) and multifocal Ewing's sarcoma (2) had received autologous (10) or allogeneic (16) BMT from related (9) or unrelated donors (7). Routine serum samples were obtained prior to BMT and at days 46 and 100 after BMT. However, in patients with severe complications additional samples were drawn at individual points in time. Prior to BMT, IL-10 serum levels were not detectable in 24/24 patients. Post-BMT, 11 patients developed elevated IL-10 levels, of these eight died of complications (DOC), whereas only one of 15 patients with undetectable IL-10 died of complications, indicating that high IL 10 levels were significantly correlated with severe life-threatening complications (chi2, P < 0.01). To determine the pathomechanism and role of the increased IL-10 levels, they were correlated to the respective NP and CRP serum concentrations. CRP and NP concentrations were found significantly elevated in patients with detectable IL-10, indicating a severe acute phase reaction associated with macrophage activation. In conclusion, high IL-10 serum levels in patients after BMT were significantly associated with fatal outcome. Since IL-10 is a strong suppressor of T cell immunity, high IL-10 production in patients with severe complications such as septic shock or GVHD > grade II after BMT might lead to functional immunodeficiency contributing to the poor prognosis of these patients. PMID- 9339751 TI - Allogeneic bone marrow transplantation for relapsed and refractory Hodgkin's disease and non-Hodgkin's lymphoma. AB - The relative benefit of allogeneic bone marrow transplantation (alloBMT) vs autologous BMT (autoBMT) for patients with relapsed or refractory Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) remains uncertain. Toxicity from graft-versus-host disease (GVHD) may diminish the potential benefits both of graft-versus-tumor activity and of receiving uncontaminated donor marrow stem cells. From 1987 to 1995, 27 adults (ages 18-60 years; median 36) underwent alloBMT for lymphoma after failure of standard chemotherapy. Twenty-one had NHL and six had HD (nodular sclerosis). Thirteen patients had primary refractory disease or chemotherapy-resistant relapses; two of these had relapsed after autoBMT. Three patients had untested relapses (one of them had relapsed after autoBMT), and 11 had chemotherapy-sensitive relapses. Twenty-four received HLA matched bone marrow from a sibling (one twin); three received haploidentical marrow cells. Nine (33%) died from lymphoma. Eleven (41%) died of treatment related causes. Opportunistic infections were a substantial problem leading to eight of these deaths (30%). Six patients (22%) survive free of lymphoma 17-70 months post-BMT (median, 56 months); four had had sensitive relapses, one had had a resistant relapse, and one had had nontested relapse. Three have chronic GVHD (limited in one; extensive in two). One HD patient who had relapsed after autoBMT remains in remission 19 months after alloBMT. No therapy-related myelodysplasia has been observed. We conclude that alloBMT has substantial morbidity in heavily pretreated lymphoma patients due to acute toxicity, infections and GVHD. However, 22% of our HD/NHL patients have had long-term disease-free survival. PMID- 9339753 TI - Varicella vaccination in children after bone marrow transplantation. AB - Herpes zoster (HZ) is one of the most common complications after bone marrow transplantation (BMT) in children. Apart from treatment with antiviral drugs, effective prevention by active immunization with varicella-zoster virus (VZV) appears to be possible. In this study 15 patients were vaccinated with a live attenuated VZV vaccine (Varilrix) 12-23 months after BMT. The vaccine was well tolerated without adverse reactions. Chickenpox or HZ were not observed for up to 2 years after immunization. Eight out of nine seronegative patients seroconverted and in six virus-specific IgG could still be demonstrated 2 years later. The incidence of VZV diseases in 133 non-immunized children after BMT was 26.3%. Infections usually occurred within 18 months after BMT. PMID- 9339752 TI - Factors influencing platelet recovery after blood cell transplantation in multiple myeloma. AB - We sought to determine factors that impact on the recovery of platelets after blood cell transplantation in patients with multiple myeloma. We performed retrospective analyses in 51 patients undergoing blood cell transplantation for multiple myeloma. The proportional-hazards model was applied to determine significant risk factors. Of 51 transplants, 14 patients failed to achieve a platelet count of 50 x 10(9)/l. Median time to a neutrophil count of 0.5 x 10(9)/l was 10.5 days. Median time to achieve a platelet count of 50 x 10(9)/l was 32 days. Multivariate analysis revealed that cyclophosphamide and G-CSF priming before collection of hematopoietic precursors (P < 0.001) was a positive predictor of rapid engraftment and prior exposure to melphalan given orally (P = 0.02) was a negative predictor of subsequent platelet engraftment. The number of mononuclear cells collected, the patient's disease status at the time of transplant and the presence of circulating plasma cells in the harvested product did not have a significant impact on time to platelet engraftment. We conclude that cyclophosphamide and G-CSF priming shortened the time to achieve platelet engraftment compared with G-CSF alone. Prior exposure to melphalan delayed platelet engraftment and can lead to complete failure of platelet recovery. Stem cells should be collected before melphalan administration in patients with multiple myeloma who are candidates for possible blood cell transplantation. PMID- 9339754 TI - Cytokine-mediated nitric oxide release--a common cytotoxic pathway in host-versus graft and graft-versus-host reactions? AB - Serial cytokine and nitrate (as a measure of nitric oxide production) levels were assayed in nine consecutive patients undergoing allogeneic haemopoietic stem cell transplants. They were compared to those in 13 patients undergoing autologous transplants (transplant controls), 15 neutropenic patients with infective complications (patient controls) and 27 blood donors (normal controls). Peak nitrate, interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF alpha) levels were significantly higher in four allogeneic transplant patients with major non-infective complications compared to those without such complications, and control groups. Cytokine and nitrate levels peaked during conditioning therapy in the patients with veno-occlusive disease (one patient) and fulminant cholestatic liver failure (one patient), indicating that tissue damage may have been initiated during chemoradiotherapy in these patients, whereas peak levels occurred 2-3 days before graft rejection (one patient) and severe graft-versus-host disease (one patient), indicating a role for cytokine induced nitric oxide release in the pathophysiology of these immune-mediated complications. Based on the data presented, it can be tentatively postulated that nitric oxide is a common proximate regulator of the immune response in host versus-graft and graft-versus-host reactions. PMID- 9339755 TI - Iatrogenic Wernicke's encephalopathy in allogeneic bone marrow transplantation: a study of eight cases. AB - Wernicke's encephalopathy (WE) is a neuropsychiatric condition generally caused by acute thiamine deficiency and classically involves the triad of altered mentation, ataxia and ophthalmoplegia. It is most common among alcoholics, but several other causes have been identified, including total parenteral nutrition (TPN) use. We present eight cases of WE in patients undergoing allogeneic BMT, where thiamine deficiency was caused by a lack of vitamin supplementation during TPN administration. Clinically, WE presented as a severe refractory metabolic acidosis, preceded by 'raspberry tongue', and ophthalmologic and neurologic dysfunction. The sites most affected were the periventricular structures and the thalamus, and no mammilary bodies lesions were found. PMID- 9339756 TI - Risk factors for hepatic veno-occlusive disease after bone marrow transplantation: retrospective analysis of 137 cases at a single institution. AB - One hundred and thirty-seven consecutive patients who received bone marrow or peripheral blood stem cell transplantation were studied retrospectively to identify the risk factors for hepatic veno-occlusive disease (VOD). Of the 137 recipients, twenty (14.6%) patients were diagnosed with VOD using the McDonald's criteria. In these 20 patients with VOD, we analyzed various clinical parameters, including age, sex, HLA status, conditioning regimen, irradiation, immunosuppressive agents, mode of transplantation, history of hepatic dysfunction, pre-transplant hepatic and renal function, infectious episodes, antibiotics use, and serum viral titers. A history of hepatic dysfunction and low levels of pseudocholinesterase before transplantation were found to be statistically significant (P = 0.04 and 0.04). Low levels of pseudocholinesterase were significant by multivariate analysis using the logistic regression model (P = 0.02). These results suggest that pseudocholinesterase levels before transplant are important markers of VOD in patients receiving BMT. PMID- 9339757 TI - Long-term follow-up of immunosuppressive treatment for obstructive airways disease after allogeneic bone marrow transplantation. AB - To describe clinical outcome with first line immunosuppression therapy for obstructive airways disease (OAD) after allogeneic BMT, we have retrospectively examined 20 long-term survivors affected by OAD. All patients had normal pulmonary function test (PFTs) before BMT. OAD was defined as FEV1 less than 80%, FER less than 80%, maximum midexpiratory flow rate of 50% vital capacity (MMFR) less than 65%, or residual volume greater than 120. Prednisone (n = 4), CsA (n = 8) and azathioprine (n = 8) have been used as first-line immunosuppression agents. Mean follow-up was 65 months (range 15-142). We identified three categories of patients according to response to treatment: complete (n = 6, 30%), partial (n = 6, 30%) or no response (n = 8, 40%). Age, FEV1, time of onset after BMT, Karnofsky index or immunosuppression modality do not seem to be related to subsequent response. However, patients with low values of MMFR and high values of RV at the beginning of therapy are likely to show poor response. In the complete response group, normalisation of PFTs is achieved within the first months of treatment (median 6 months ranging from 3 to 9 months), suggesting that prolonged therapy is not advantageous and could increase morbidity and mortality if there are no other signs of CGVHD. PMID- 9339758 TI - CD34+ cell selection in chronic phase chronic myeloid leukaemia: a comparison of laboratory grade columns. AB - CD34 positive (CD34+) cell selection is increasingly used for a number of important applications including gene therapy studies, ex vivo expansion and purging. However there are no data regarding the use of different technologies for CD34+ cell selection in chronic myeloid leukaemia (CML). We therefore compared the performance of three laboratory grade CD34+ selection columns (MiniMACS, Cellpro Ceprate LC and Baxter Isolex 50), using CML chronic phase peripheral blood (PB) and bone marrow (BM). With different CML samples the CD34+ purity from the three columns was equivalent, but comparing five paired samples the Ceprate purity was greater than MiniMACS, at 92.5 and 80.9%, respectively, P = 0.04. Combining results from paired and unpaired CML samples, MiniMACS (n = 7) gave a higher CD34+ yield than Ceprate LC (n = 8) or Isolex 50 (n = 4) with a mean of 51.1%, 24.3% and 13.2% respectively, (P = 0.04 and 0.01). Cell losses with all columns were similar. Attempts to improve the yield from the Ceprate LC columns by modifying the method were unsuccessful. Following MiniMACS and Ceprate LC separation the clonogenic potentials of CD34+ cells in the pre- and positive cell fractions were the same. The proportion of CD34+ 38- or CD34+ DR- cells was unchanged following column separation. These data suggest that the MiniMACS column may be the best column for CD34+ cell selection in CML but these results must be confirmed using large scale clinical columns once the MiniMACS column is licensed. It is possible that variations in CD34+ cell yields between the different columns reflect differences in antibody binding affinity to CML cells, or differences in column technologies. PMID- 9339759 TI - Combined positive/negative selection for highly effective purging of PBPC grafts: towards clinical application in patients with B-CLL. AB - Autologous PBPC transplantation is a potentially curative treatment for patients with chronic lymphocytic leukemia (CLL). As the autografts are frequently contaminated with large numbers of tumor cells, we have studied double purging of PBPC using immunomagnetic CD34+ cell selection (Isolex 300i) followed by negative depletion with anti-CD19/20/23/37-labeled immunomagnetic beads. In four small scale experiments using PBPC from patients with CLL or leukemic low-grade lymphoma, double purging resulted in CD34+ enrichment from 0.9-4.4% to 95.8 99.4%. Lymphoma cells were always undetectable by FACS and PCR (CDR3 or t(14;18)) after negative depletion. Next, seven heavily contaminated full grafts from five patients with CLL or lymphoplasmocytoid immunocytoma were subjected to the double purging procedure, resulting in a CD34+ enrichment from 1.6% (0.7-5.6) to 98.5% (96-99.8). The CD34+ yield after double purging was 1.3-6.3 x 10(6)/kg, according to a median recovery of 32%. The overall reduction of lymphoma cells was 5.6 (>4.6-6) log. Although CLL cells were completely absent after purging in five cases as assessed by FACS, all grafts remained PCR positive. The first two patients have been reinfused with double selected products after myeloablative radiochemotherapy and showed prompt and uneventful hematopoietic engraftment. We conclude that without significant loss of CD34+ cells, negative depletion adds 2 log of CLL cell depletion to CD34+ selection, resulting in an overall purging efficacy of more than 5 log. This combination of positive and negative selection can be successfully applied even to heavily contaminated PBPC grafts. PMID- 9339760 TI - Allogeneic transplantation after a conditioning regimen with ifosfamide, carboplatin and etoposide (ICE). AB - Successful allogeneic hematopoietic transplants require conditioning regimens with sufficient immunosuppression to allow acceptance of the allograft. Cyclophosphamide, in combination either with TBI or with chemotherapeutic drugs, is the keystone of commonly used regimens. The toxicities of TBI and tumor resistance to cyclophosphamide create a niche for alternative, chemotherapy-based conditioning regimens. We report successful allogeneic stem cell transplantation after an ifosfamide-based regimen with ifosfamide 20 g/m2, carboplatin 1.8 g/m2 and etoposide 3 g/m2 (ICE) in divided doses over 6 days. Engraftment was prompt with neutrophils > or = 20 x 10(9)/l on day +10 and platelets > 20 x 10(9)/l on day +18. Engraftment of donor cells was documented by chromosome analysis and by VNTR analysis. An ifosfamide-based regimen provides sufficient immunosuppression for hematopoietic allograft acceptance in the absence of cyclophosphamide or of TBI. PMID- 9339761 TI - Disseminated nocardiosis after allogeneic bone marrow transplantation. AB - Nocardia infarction is rare in bone marrow transplant recipients; only 10 cases have been reported. We describe a case of disseminated nocardiosis after allogeneic BMT with positive blood cultures and multiple cerebral and hepatosplenic abscesses. PMID- 9339762 TI - Treatment of severe Evans syndrome with an allogeneic cord blood transplant. AB - Immunosuppressive therapy is commonly used in the management of autoimmune disorders. As marrow-derived lymphocytes appear to play a key role in these diseases, lymphoid ablation followed by replacement with autologous or allogeneic stem cells may be a therapeutic option. We report a 5-year-old boy with severe Evans syndrome which consists of immune thrombocytopenia and Coombs-positive hemolytic anemia. He was rendered into complete remission with marrow ablation followed by rescue with an HLA-identical sibling cord blood transplant. He unexpectedly died 9 months following transplant from acute hepatic failure of unknown etiology. PMID- 9339763 TI - Child health and the Cochrane Collaboration. PMID- 9339765 TI - Peer relations and friendship in physically disabled children. AB - This article presents the results of a literature study and qualitative fieldwork into the way physically disabled children experience friendship. We discuss literature on how children make contact with peers, on the nature of their friendship and on what it is like to be excluded. Some aspects are complemented with information from the children themselves. Peer relations and friendship are the very essence of a happy childhood. This article therefore closes with a number of recommendations to improve peer relationships. PMID- 9339764 TI - Parental separation: effects on children; implications for services. AB - It is commonly assumed that the children of separated or divorced parents are disadvantaged financially and in terms of social development. A small-scale study was designed involving 14 volunteer parents who were separated from their spouses/partners and their 28 children, ranging in age from 5 to 12 years. Structured interviews examined the families' attitudes and feelings about the separation, the mental health of the parents, the emotional and behavioural reactions of the children, and the families' use of services. The results suggested that over 60% of the girls showed internalizing, and over 60% of the boys displayed externalizing behavioural difficulties. About 40% of the children suffered psychosomatic disorders. Nearly 75% of the parents suffered depression before and after separation. Depression, itself, may act to exacerbate children's emotional and behavioural difficulties. Although all the parents had received medication for their depression, few of the children had received any help for their difficulties. The urgent need for services to help such children is discussed. PMID- 9339766 TI - A re-appraisal of screening for colour vision impairments. AB - Screening for colour vision impairments (CVI) has been carried out in schools in the UK since 1934, but little is known about its yield or value. A survey was conducted among Sheffield schoolboys, school nurses and optometrists to determine the benefits of CVI screening. The results indicated that between 4.2 and 5.2% of boys had been identified as having a CVI, compared with the expected prevalence of 8%. Boys were ill-informed about the significance of CVI for careers planning but recognized the potential importance of having this information before making decisions about choice of subjects and examinations. Possible reasons for the low yield of screening are reviewed and alternative strategies are discussed. PMID- 9339767 TI - Evaluation of a cancer-based coping and caring course used in three different settings. AB - Cancer in the family may affect the psycho-social adjustment of the children involved. Children who have survived childhood cancer or with a parent or sibling with cancer, may find protection from the risk of emotional and behavioural difficulties from effective external support systems. A training course was developed by the Cancer Research Campaign to help professionals who make up potential external support systems, cope with children experiencing cancer related life crises. The course was offered to, and evaluated with, practising teachers, student teachers and Cancer Aid and Listening Line workers. Results from interviews and pre and post course questionnaires indicated that after the course all three groups felt more confident in dealing with children under stress. Participants felt that specific skills such as 'attentive listening' had increased as a result of the course. Of the three groups the practising teachers have found the course the most useful in their work and have been able to disseminate their newly acquired skills throughout their schools. Teachers are expected to cope with students' various life crises as they arise in schools. Basic counselling skills training for teachers could help them cope with these in a more confident manner, thereby reducing the risk of emotional and behavioural difficulties and contributing towards a Health Promoting School ethos. PMID- 9339768 TI - Perceptions of the risk of child abduction or loss and the utility of child electronic security devices. AB - Perceptions of the susceptibility of young children to becoming lost or being abducted, and of the potential usefulness of child electronic security devices, were examined via a questionnaire. Data were provided by 41 volunteers, most of them from a local government office centre. The questionnaire asked for demographic data, and then for the risk of a child being abducted or lost when under the supervision of different caregivers and in different situations. The probable effectiveness of three common abductor ploys was also addressed. The questionnaire concluded with 10 questions about child electronic security devices. Respondents viewed mothers, fathers, and grandparents as equally responsible caregivers and young adults/babysitters as the least responsible. These effects diminished as the age of the children increased. The garden at home was judged to be the most secure environment for children of all ages, while an amusement park was judged the least secure environment. Children were perceived to be more at risk of an abduction when a stranger asked for physical assistance or to take them to the hospital because their parents were hurt, than when asked for directions. Furthermore, the respondents expressed a moderately strong need for child electronic security devices, and viewed parents who use them as more responsible than those who do not. PMID- 9339769 TI - Urinary antimony levels in infants are low and unrelated to age or passive smoking. PMID- 9339770 TI - Delayed motor milestones are relatively poor predictors of cerebral palsy in high risk pre-term infants. PMID- 9339771 TI - Evaluating the consequences of hospital restructuring. PMID- 9339772 TI - Hospital restructuring in North America and Europe. PMID- 9339773 TI - Hospital restructuring in the United States, Canada, and Western Europe: an outcomes research agenda. AB - OBJECTIVES: This article describes the extent and nature of hospital restructuring across the United States, Canada, and Western Europe, countries with differently organized and financed health-care systems, and assesses the feasibility of international research on the outcomes of hospital restructuring. METHODS: The conceptual background, context, and focus for the Bellagio conference on Hospital Restructuring in North America and Western Europe held in November 1996 is provided, illustrating key issues on hospital and workforce trends using the US data with international comparisons. RESULTS: Hospital systems internationally are undertaking very similar restructuring interventions, particularly ones aimed at reducing labor expenses through work redesign. Nursing has been a prime target for work redesign, resulting in changes in numbers and skill mix of nursing staff as well as fundamental reorganizing of clinical care at the inpatient unit level. Yet little is known about the outcomes of such organizational interventions and there are few efforts in place to critically evaluate these actions. CONCLUSIONS: Restructuring of the hospital workforce and redesign of work in inpatient settings is widespread and markedly similar across North American and Europe, and warrants systematic study. Cross-national studies of the impact of restructuring inpatient care on patient outcomes would yield valuable lessons about the cost-quality tradeoffs in hospital redesign and re engineering, as well as inform national planning about the numbers and types of nurses needed in the coming decades. PMID- 9339774 TI - The reform of hospital care in the Netherlands. AB - OBJECTIVES: This article provides a short overview of the structure of hospital care in the Netherlands. It discusses a number of hospital reform programs that have been started since the early 1980s and others more recently proposed. Attention has also been given to the potential impact of hospital reform. METHODS: Descriptions of hospital structure, planning, financing, payment of medical specialists, and hospital workforce and services provide a context for a discussion of current hospital reform programs. Trends in hospital care delivery, strategic management, and internal organization are examined. RESULTS: The relationships between the hospital and other health-care providers, health insurers, employers, and patients are found to have a significant impact on the future of the hospital. The interconnections of these four factors will help form the basis for an understanding of ongoing hospital change. CONCLUSIONS: Hospital care has been subject to rapid change during the last two decades, as hospitals become centers for acute specialized medical care. This development has important consequences for the position of hospitals in the health-care delivery network, as they become one of a number of providers in a more integrated delivery system. It should be noted that the changes in hospital care cannot be understood as the results of a single reform program, but rather as the result of a long series of reform efforts. PMID- 9339775 TI - Financing reforms in the German hospital sector: from full cost cover principle to prospective case fees. AB - OBJECTIVES: The authors provide an overview of the hospital sector in Germany with a focus on the impact of recent reform legislation on this sector. METHODS: Data from the Federal Statistics Office, the Ministry of Health, and the Federal Association of Physicians are synthesized with information obtained from a general review of the literature. RESULTS: Before the implementation of recent health-care reforms, the German health-care system has been sharply divided into inpatient and ambulatory care sectors, resulting in a fragmented system of care delivery. All hospital operating costs were fully covered through per diem charges. The 1992 Health Care Structure Act and subsequent pieces of legislation have introduced new mechanisms to improve cost efficiency in the hospital sector and increase coordination between the inpatient and outpatient care. These measures notably include implementing an inpatient prospective payment system and permitting ambulatory surgery and care services to be offered in inpatient settings. CONCLUSIONS: Whereas prospective payments have greatly reduced the length of stay, hospitals were reluctant to offer ambulatory surgery due to budgetary constraints and the high level of ambulatory surgery by office-based physicians. The reforms passed have not yielded substantial cost savings. These reforms offer a natural experiment that could benefit from national and international studies on the impact of hospital sector redesign on management, financing, and patient outcomes. PMID- 9339776 TI - Hospitals in England: impact of the 1990 National Health Service reforms. AB - OBJECTIVES: This article aims to describe recent changes in English hospitals, with particular reference to the impact of the National Health Service (NHS) and Community Care Act of 1990. METHODS: Significant policies that have affected the functioning of the hospital sector of the British NHS are reviewed. Data from the NHS Department of Health are used to describe trends in utilization. RESULTS: The NHS and Community Care Act of 1990 radically changed the financial and organizational framework within which hospitals operate. By creating separate purchasing organizations, the act opened the way for competition between hospitals. In practice, such competition has been very limited. Central directives aimed at reducing waiting times for nonurgent admissions, as well as at raising the volume of work done relative to the finances available have been more significant influences. These changes, combined with rising numbers of emergency admissions, have put the physical and human resources of English hospitals under intense pressure. Admissions have risen, lengths of stay have fallen across all age groups, and ambulatory care has grown rapidly. CONCLUSIONS: There is little consensus on the future direction regarding the role and structure of acute-care hospitals. There is evidence, though, that improvements in the process and outcomes of care are possible within the current financial and organizational framework of the hospital sector. PMID- 9339777 TI - Hospital restructuring initiatives in Canada. AB - OBJECTIVES: Recent changes in the organization, staffing, and utilization of acute hospitals in Canada are reviewed with regard to the potential implications for quality of care, national nurse workforce requirements, and research. METHODS: Available national and selected provincial data and trends in hospital utilization, capacity, and staffing are synthesized. RESULTS: Health system reform in Canada has resulted in lower utilization of acute inpatient resources, excess hospital capacity, and increased budgetary constraints in the hospital sector. In response, there is widespread hospital restructuring, which includes modifications in nurse staffing ratios and skill mix. Little is known about the potential impact of these changes on patient outcomes. From a workforce perspective, changes in the hospital sector have reduced demand for registered nurses but nursing schools have not modified enrollments. As a result, new graduates are experiencing difficulty obtaining registered nurse positions. CONCLUSIONS: Research should be undertaken to evaluate the impact of changes in the organization and staffing of hospitals on patient outcomes, and on the future requirements for nurses. PMID- 9339778 TI - Canadian hospitals in transformation. AB - OBJECTIVES: This article reviews health system reform in Canada and discusses how hospitals are restructuring staffing patterns and redesigning work in response to increasing budgetary constraints. METHODS: Two models for work redesign at the patient unit level are contrasted, one employing a mix of registered nurses and registered practical nurses and one using registered nurses and assistive personnel. Principles and rationale for work redesign are discussed. RESULTS: Both models for patient care redesign reduce registered nurse positions and both appear to yield some cost savings. These models for patient care redesign are controversial among providers, and there is no consensus as to which model is preferable. No research has been undertaken to determine whether staffing changes and work redesign result in adverse patient outcomes. CONCLUSIONS: Regionalization of hospitals is reducing inpatient capacity in Canada, although the pace of regionalization varies in the different provinces. Hospital re engineering is designed to reduce expenditures by reducing the cost of staffing. Research is needed to evaluate the results of re-engineering on patient outcomes and their relationship to registered nurse staff reductions. PMID- 9339779 TI - A spurious correlation between hospital mortality and complication rates: the importance of severity adjustment. AB - OBJECTIVES: When two outcome measures, such as mortality and complication rates, are intended to measure the same underlying quantity (in this case hospital quality of care), one expects they will be highly correlated. In addition, as data quality improves, one expects the correlation will increase. The authors show that these expectations are, in a significant way, mistaken. METHODS: The authors study two outcomes (hospital mortality and complication rates after surgery) using three predictive models that vary in adjustment for severity of illness. RESULTS: Two hospital rankings, based on each of the two outcomes, are well correlated when not adjusted for severity. However, as clinical data are added to the models, the correlation tends to disappear. The authors explain this based on assumptions regarding the relative size of the partial correlations between mortality, complication rate, and severity covariates. CONCLUSIONS: Before claims of construct validity can be made, investigators must show that correlations between outcomes purporting to measure quality of care are sustained after adequate correction for severity. Most importantly, it should be recognized that inadequately controlled confounding variables may lead to a spurious high correlation between an accepted and a new outcome measure, and a false sense of adequate construct validity. PMID- 9339780 TI - Hospital restructuring and the epidemiology of hospital utilization: recent experience in Ontario. AB - OBJECTIVES: The author highlights changes in hospital utilization that have occurred in association with restructuring of Ontario hospitals. The basic features of the epidemiology of hospital utilization described link the analysis of the organizational and structural components of hospitals with a more comprehensive evaluation of the impacts of their restructuring and have implications for international comparative studies. METHODS: Data from the Canadian Institute for Health Information and the Canadian census were analyzed to provide a population-based description of hospital utilization and care. These hospital data provided information on changes in the patterns of care that occurred during restructuring, based on hospital separations for the fiscal years 1991-1992 through 1995-1996. RESULTS: Analysis of hospital utilization patterns revealed a 30% decrease in the days of care provided per 1,000 population during the period, the result of declines in both the age-adjusted inpatient separation rates and average length of hospital stay. The shift of surgical treatment to outpatient settings contributed to the reduction in inpatient days of care. The decline in utilization was experienced unevenly across age groups, with the elderly experiencing less of the decline than did younger age groups. Individuals living in the poorest areas used more inpatient care than did those living in the richest areas, although the gap in utilization narrowed over the period. CONCLUSIONS: International comparisons of the epidemiology of hospital utilization and the impact of hospital restructuring will require the use of multiple data sources and the development of shared evaluative frameworks. Health data systems in Canada support the assessment of the broader impacts of hospital restructuring and offer a framework for developing research projects that can provide useful information on these important changes in health-care policy. PMID- 9339781 TI - Using routine data to evaluate quality of care in British hospitals. AB - OBJECTIVES: As part of the process of developing an international comparative study of the outcome of hospital care it has been necessary to understand what data on hospital activity are available and how they compare with those in other countries. The authors describe the administrative data collected in British National Health Service Hospitals, with emphasis on how they differ from that available in the United States. METHODS: A description of the content of administrative data and a selective review of published literature related to their uses. RESULTS: Administrative data in the United Kingdom resembles that available in the United States, but it also has some important differences. These include a different system of procedure classification, the use of International Classification of Diseases, 10th Revision (rather than International Classification of Diseases, Ninth Revision, Clinical Modification), and the use of a different denominator ("finished consultant episodes," rather than admission spells). There also are important differences in the completeness and precision of the data that are collected. CONCLUSIONS: British administrative data can be used to describe patterns of care and have the potential to identify possible differences in outcome that can then be subjected to more detailed scrutiny. They are substantially less detailed than US data but are also somewhat less costly to collect. PMID- 9339782 TI - Hospital outcomes research in Germany: results from a retrospective survey among sickness fund beneficiaries. AB - OBJECTIVES: The authors assess the feasibility of using retrospective, indication specific patient surveys to conduct hospital outcomes research in Germany. Surgical outcome and patient satisfaction were examined in patients who underwent common elective surgical procedures. METHODS: Using the International Classification of Diseases Ninth Revision coding available in the Schwabisch Gmund health insurance data base, all patients for a defined period of time with one of the three following diagnoses were selected and questioned retrospectively using an indication-specific survey instrument: (1) varicose veins of the lower extremity; (2) nasal septum deviation; and (3) inner knee joint damage limited to patients undergoing arthroscopic meniscus repair. Survey content focused on preoperative conditions, pre- and postoperative symptoms, postoperative complications, the nature and duration of postoperative follow-up, and satisfaction with surgical outcome. RESULTS: Significant postoperative improvement of preoperative symptoms was found for all three groups. Complete freedom from symptoms was found in 29.7% of patients treated for varicose veins, 24.1% of patients with meniscus repair, and in only 10.6% of patients with nasal septum deviation. Multivariate analyses indicated that postoperative impairment was the decisive variable governing patient satisfaction for all three groups. CONCLUSIONS: The use of retrospective, indication-specific patient surveys constitutes a time-efficient, cost-effective, and patient-focused option for the systematic acquisition and evaluation of health outcomes in Germany. This methodology holds promise for international and domestic efforts to demonstrate the consequences of restructuring activities in the inpatient sector. PMID- 9339783 TI - Hospital reform and nursing labor market trends in Canada. AB - OBJECTIVES: Trends in the Canadian registered nurse (RN) workforce during the past 3 decades are examined, and the implications of current hospital sector retrenchment for RN employment are considered. METHODS: A descriptive review using relevant literature and existing databases on the nurse workforce is presented. RESULTS: From the 1960s through the 1980s, the Canadian RN workforce grew exponentially, fueled by expansions in the health-care delivery system under Medicare, rising inpatient acuity and skill-intensive patient care, enhanced access to nursing education, and increases in the numbers of women entering the workforce. Acute care hospitals have and continue to be the predominant employer of RNs. However, the 1990s have witnessed considerable hospital retrenchment, and with that retrenchment the growth of the hospital RN workforce has slowed dramatically. CONCLUSIONS: The ultimate outcomes of hospital retrenchment for the RN workforce remain unclear. Some speculate that the quality of care and working conditions will deteriorate in hospitals, as hospital administrators replace RN staff with lesser trained personnel to reduce costs. Others see change in the hospital sector as an opportunity for RNs to expand their scope of practice and responsibility in outpatient settings. The need for national and international research on the outcomes of hospital restructuring on patient care and the work of RNs is critical to sound policy making. PMID- 9339784 TI - Trends in hospital restructuring and impact on the workforce in Germany. AB - OBJECTIVES: The author describes the impacts of hospital restructuring and reform legislation in Germany on the nursing workforce. METHODS: A descriptive analysis using selected literature is presented. RESULTS: Driven by the increased service needs of an aging population and the imperative to contain health-care costs, the hospital sector is shrinking while increasing its intensity of care delivery. Within this environment, the demand for patient-focused, high-quality nursing care is high, whereas the number of new graduates entering the nursing field is declining. Despite absolute increases in the number of nurses employed by the hospital sector, evidence suggests that hospitals are operating with a nursing workforce deficit. The recent reform law of 1992 mandates several changes with large implications for nursing. These include a linking of the hospital sector with outpatient care; a focus on rigorous, interdisciplinary quality assurance; and a revaluing of the adequacy of hospital nurse staffing. CONCLUSIONS: Hospitals will remain the major employers of nurses, with new outpatient sector opportunities. Adequate nurse staffing methodologies, sound personnel retention strategies, and reform of care delivery models are needed to assure high-quality nursing care in the hospital sector. PMID- 9339785 TI - Health sector reform and trends in the United Kingdom hospital workforce. AB - OBJECTIVES: The authors examine changing trends in the profile and patterns of employment of the workforce in hospitals in the National Health Service (NHS) in the United Kingdom. The effect of the implementation of the NHS reforms is considered, with particular reference to the changing composition of the nursing workforce. The authors note that there are problems with establishing trend data because of altered information requirements as a result of the NHS reforms. METHODS: Analysis and review of data from secondary sources and research publications. RESULTS: Although hospital activity rates have grown, patient length of hospital stays decreased, and patient activity levels increased, there has not been a linked growth in the size of the nursing workforce. The main changes in the profile of the nursing workforce highlighted are a marked reduction in the numbers of nursing students and alterations in the skill mix between first- and second-level qualified nurses. The authors also note a large increase in the number of managerial and administrative staff employed and growth in medical staff numbers. Changes in working patterns and increases in contracting for support services and in the use of temporary staff also are discussed. CONCLUSIONS: There have been pronounced changes in the profile of the hospital workforce but little evaluation of the impact of these changes on outcomes of care. PMID- 9339786 TI - Posturographic evidence of nonorganic sway patterns in normal subjects, patients, and suspected malingerers. AB - During the last 10 years, computerized dynamic posturography has yielded various patterns of sway on the sensory organization test and the motor control test that have been associated with a variety of organic balance disorders. Some aspects of performance during computerized dynamic posturography, however, are under conscious control. Voluntary movements not indicative of physiologic response to balance system stimulation can also affect computerized dynamic posturography results. Quantification of nonorganic or "aphysiologic" response patterns in normal subjects, patients, and suspected malingerers is crucial to justify use of computerized dynamic posturography for identification of physiologically inconsistent results. For this purpose the computerized dynamic posturography records of 122 normal subjects, 347 patients with known or suspected balance disorders, and 72 subjects instructed to feign a balance disturbance were critically evaluated by use of seven measurement criteria, which were postulated as indicating aphysiologic sway. Each criterion was scored with a standard calculation of the raw data in a random, blinded fashion. The results of this multicenter study show that three of the seven criteria are significantly different in the suspected "malingerer" group when compared with either the normal or patient group. The relative strength of each criterion in discerning organic from nonorganic sway provides the examiner with a measure of reliability during platform posture testing. This study demonstrates that computerized dynamic posturography can accurately identify and document nonorganic sway patterns during routine assessment of posture control. PMID- 9339787 TI - Refinement in the rehabilitation of the paralyzed face using botulinum toxin. AB - A number of surgical procedures exist to improve facial symmetry for patients with facial paralysis. Whereas static symmetry is often improved, dynamic asymmetry frequently persists because of the imbalance of complex coordinated movements of facial expression. The paralyzed face is often distorted by the excessive pull of the normal contralateral face during emotional expression. This study reports an expanded clinical indication for botulinum toxin in patients with unilateral facial paralysis. Ten patients with facial paralysis and markedly asymmetric smiles were treated with botulinum toxin A injections into the contralateral zygomaticus major, levators labii superioris and angulii oris, or risorius muscles. Eight of the 10 patients noted improvement in the symmetry of their smiles and underwent repeat injections. The onset and duration of effect averaged 5.9 days and 3 months, respectively. Botulinum toxin therapy provides a safe and efficacious modality for refining the appearance of the paralyzed face during mimetic activity. PMID- 9339788 TI - Hemifacial spasm: endoscopic vascular decompression. AB - Sixty patients with primitive hemifacial spasm were treated by means of a minimally invasive retrosigmoid approach in which endoscopic and microsurgical procedures were combined. Intraoperative endoscopic examination of the cerebellopontine angle showed that for 56 of the patients vessel-nerve conflict was the cause of hemifacial spasm. The most common offending vessel was the posterior inferior cerebellar artery (39 patients), next was the vertebral artery (23 patients), and last was the anterior inferior cerebellar artery (16 patients). Nineteen of the patients had multiple offending vascular loops. In one patient, another cause of hemifacial spasm was an epidermoid tumor of the cerebellopontine angle. For three patients, it was not possible to determine the exact cause of the facial disorder. Follow-up information was reviewed for 54 of 60 patients; the mean follow-up period was 14 months. Fifty of the patients were in the vessel-nerve conflict group. Forty of the 50 were free of symptoms, and four had marked improvement. The overall success rate was 88%, and there was minimal morbidity (no facial palsy, two cases of severe hearing loss). PMID- 9339789 TI - Impairment and disability in patients with facial neuromuscular dysfunction. AB - Integration of impairment measures and disability measures may provide clinicians with an accurate and comprehensive picture of the patient's dysfunction. The purpose of this study was to indicate the usefulness of two new scales of measuring facial impairment and disability in describing characteristics of individuals with facial neuromuscular dysfunction. Fifty-one individuals with unilateral facial neuromuscular dysfunction and a House-Brackmann grade of III or higher were included in the study. The subjects' movement impairments were assessed using the Facial Grading System (FGS). The subjects reported their physical and social function on the Facial Disability Index (FDI). Nine variables were subjected to a confirmatory principal-components factor analysis to indicate important factors in describing patients with facial nerve disorders. The confirmatory principal-components factor analysis identified three factors, impairment, disability, and temporal characteristics of disease, accounting for 72% of the variance in describing individuals with facial neuromuscular dysfunction. Integration of these measures may provide clinicians with an accurate and comprehensive picture of the patient's dysfunction, thus aiding in the determination of intervention and the measurement of clinical outcomes. PMID- 9339790 TI - Comparative value of facial nerve grading systems. AB - The Fisch Detailed Evaluation of Facial Symmetry (DEFS) and House-Brackmann grading system (HBGS) were compared by statistical examination for their reliability and interobserver variability. Furthermore, the correlation and agreement with a standard global evaluation were compared. Therefore 47 patients with facial palsy of different cause have been evaluated with the two systems, and the global overall evaluation was done by five otolaryngologists familiar with facial palsy. The DEFS showed a high reliability of 0.93 compared with a reliability of 0.77 with the HBGS (international standard requires a reliability of at least 0.8). The mean interobserver variability is 5.24% (SD = 3.2%) with the DEFS and 9.26% (SD = 5.0%) with the HBGS; with a confidence interval of 95%, it is 11.6% and 19.26%. The correlation of both gradings with the global evaluation was high, with r = 0.98 and r = 0.97. The DEFS shows an excellent agreement with the global overall evaluation in 41 (87%) of 47 cases and the HBGS in 32 (66%) of 47 cases. PMID- 9339791 TI - Gender and racial variations in cephalometric analysis. AB - Cephalometric analysis has earned a vital role in the evaluation of obstructive sleep apnea. However, the normal measurements cited in the literature are not sex or race specific. Skeletal differences in sexes and races have been established. This study was initiated to examine whether race and sex variations in soft tissue and skeletal measurements exist in cephalometric analysis. A total of 89 volunteers of different race and sex participated in this study. The data support the hypothesis that there are statistically significant differences in (1) sella nasion-subspinale angle (SNA) between black men, and both Caucasian and Hispanic men, (2) sella-nasion-supramentale angle (SNB) between black men and Caucasian men, (3) posterior airway space between Caucasian men and women, and (4) mandibular plane to hyoid distance between Caucasian men and women. These data suggest that only SNA and SNB need racial specificity. Furthermore, Caucasian women need a separate set of normal values from men, specifically posterior airway space and mandibular plane to hyoid bone. PMID- 9339792 TI - Conservative subtraction-addition rhinoplasty. AB - The deviated external nose remains a difficult technical challenge to even the most masterful rhinoplastic surgeon. The classic septorhinoplasty approach to the deviated nose demonstrated a 9.8% revision rate in this study. During a 2-year period, the conservative subtraction-addition rhinoplasty procedure was developed, which subsequently reduced the revision rate to 1.3%. Conservative subtraction-addition rhinoplasty foregoes aggressive septal surgery and equalizes and enhances the airway through asymmetric turbinate volume reduction. Minimal bony and upper lateral work through rasping, soft tissue removal, and/or cartilage grafting allows for external nasal alignment. Both internal and external conservative subtraction-addition rhinoplasty components thus maintain perioperative structural stability and ensure long-term nasal symmetry. PMID- 9339793 TI - Preoperative and postoperative nasal septal surgery assessment with acoustic rhinometry. AB - INTRODUCTION: Acoustic rhinometry is a relatively new tool used for the measurement of the geometry of the nasal fossa. We hypothesized that acoustic rhinometry would be useful for preoperative and postoperative assessment of patients undergoing septal surgery. METHODS AND MATERIAL: Twenty-four patients undergoing septal surgery performed by two surgeons underwent preoperative and postoperative rhinometry. The indications for surgery were nasal obstruction caused by a deviated nasal septum. Rhinometry was conducted with the Eccovision Acoustic Rhinometry System (Hood Laboratories). Analysis of the data was performed with the Kwikstat program (Texasoft) and Excel (Microsoft). RESULTS: Subjective improvement in nasal patency was significantly correlated with improvement in acoustic rhinometry. CONCLUSIONS: Acoustic rhinometry is valuable in objectively confirming nasal patency after nasal septal and turbinate surgery. PMID- 9339794 TI - The assessment of nasality with a nasometer and sound spectrography in patients with nasal polyposis. AB - With the development of computerized acoustic analysis systems, an objective measure of nasal speech has become readily available by means of a simple, noninvasive technique. In this study, we assessed the nasality in patients with multiple nasal polyposis before and after endoscopic sinus surgery. With the nasometer, we measured nasalance, which reflects the ratio of acoustic energy output of nasal sounds from the nasal and oral cavities, and the slope score of the nasogram curve. The nasalance scores of nasal sentences and the slope scores of the nasogram curves for all nasal consonants were significantly lower in patients with nasal polyposis than in healthy subjects. After surgery, however, the nasalance and slope scores increased significantly to the normal range. On the sound spectrographic analysis, the frequencies of the first nasal formant decreased slightly and the sound intensity increased slightly for all nasal consonants after surgery. However, no significant change was noticed in the frequencies of the second nasal formant. In conclusion, nasometric and sound spectrographic analyses are considered to be useful tools for objectively assessing the extent of nasality in patients with nasal airway obstruction. PMID- 9339795 TI - A comparison of the nasal cross-sectional areas and volumes obtained with acoustic rhinometry and magnetic resonance imaging. AB - Acoustic rhinometry (AR) evaluates the geometry of the nasal cavity with acoustic reflections and provides information about nasal cross-sectional areas (CSA) and nasal volume within a given distance. The accuracy of the information obtained by AR was compared with that of magnetic resonance imaging (MRI) of the nasal cavity. Five healthy subjects were evaluated with AR and the MRI before and after the application of a long-acting nasal decongestant spray, to eliminate possible interference of the nasal cycle with both measurement techniques. The MRI images of 2 mm coronal sections of the nasal cavity were traced by three independent observers and the CSAs were calculated by computer-aided imaging digitization, to be compared with the calculated CSAs obtained with the AR at the corresponding distance from the nasal tip. Digitized data from the MRI images were also used to calculate the nasal volume within the first 6 cm from the nasal tip and compared with the AR volume measurements. The interobserver variation of digitized MRI data predecongestant and postdecongestant was not significant. The correlations of CSA and volume measurements between the AR and MRI were high (0.969) after the application of the decongestant. The correlation between the AR and MRI measurements before the decongestant was low (0.345). This may have been the result of interference of the nasal cycle during the long MRI measurements (1 hour) or other unknown factors. We conclude that AR measurements of nasal CSAs and volumes provide accurate information when compared with the MRI of the decongested nasal airway. PMID- 9339796 TI - Diffuse nasal polyposis: postoperative long-term results after endoscopic sinus surgery and frontal irrigation. AB - Diffuse nasal polyposis remains a challenge despite recent improvements in endonasal surgery. The purpose of this study is to evaluate the results after a radical complete sphenoethmoidectomy with peroperative and postoperative frontal irrigation in cases of diffuse nasal polyposis. In this prospective study, we include 50 consecutive patients with diffuse nasal polyposis suffering from nasal obstruction, anosmia, and other symptoms of chronic sinusitis. All patients were refractory to medical therapy. In each patient an endoscopic complete sphenoethmoidectomy including total excision of all diseased ethmoid mucosa was performed. Preoperative and postoperative frontal irrigation was performed systematically. The patients were followed closely with serial endoscopic examination, and CT scanning was performed between 2 and 3 years after surgery. There were no complications. Thirty-nine of the 50 patients regained satisfactory olfaction. Partial nasal obstruction persisted in four of the 50 patients. Endoscopically, polyp recurrence was noted in 3% of posterior ethmoids, 23% of anterior ethmoids, and 50% of frontal recesses. We conclude that in cases of refractory and extensive nasal polyposis, a total sphenoethmoidectomy with perioperative frontal irrigation followed by long-term postoperative topical steroid therapy provides excellent improvement or cure with safety and reliability. PMID- 9339797 TI - Sinusitis in the intensive care unit patient. AB - A retrospective study was undertaken to determine the incidence of sinusitis as a source of fever in the intensive care unit (ICU) patient, evaluate the effectiveness of radiologic studies in diagnosing sinusitis, and develop guidelines that may help predict the result of antral lavage. Sixteen of 52 (30.7%) lavages in patients studied with plain films and 27 of 67 (40.3%) lavages in patients studied with computed tomography of the sinuses revealed purulence in the maxillary sinuses. Conversely, 23 of 30 (76.7%) of the cases with purulence in the middle meatus had purulence in the maxillary sinus (chi-squared = 27.1). If no purulence was seen, the results of the antral lavage were negative in 68 of 89 cases (76.4%). When physical examination was used in conjunction with computed tomography, 92.3% of lavages confirmed purulence in the maxillary sinus (chi squared = 16.6). In conclusion, the most important factor in predicting a positive result with antral lavage is the presence of purulence in the middle meatus on physical examination in conjunction with the presence of sinus disease on computed tomography of the sinuses. PMID- 9339798 TI - Immunotherapy for allergic fungal sinusitis: the second year. AB - Since August 1994 we have followed a protocol of treating patients with histologically proven allergic fungal sinusitis with surgical extirpation of the involved sinuses, followed by immunotherapy using both fungal and nonfungal antigens to which hypersensitivity is demonstrated by in vitro and skin testing methods. Despite predictions to the contrary, we have encountered no evidence that these injections have worsened the condition of any patients. Rather, we have noted a marked decrease in nasal crusting in all patients, with a minimum amount of recurrent polypoid mucosa and a lessened or absent requirement for corticosteroids (systemic or topical). Two patients treated with immunotherapy required systemic corticosteroids and subsequent revision surgery for residual disease that was present before the start of immunotherapy, and they have done well since. Our experience indicates that the triad of adequate surgery, frequent follow-up and medical management, and immunotherapy with relevant fungal and nonfungal antigens represents an effective means of treating patients with allergic fungal sinusitis. Nevertheless, an even longer period of study will be necessary to provide the final answer regarding the role of immunotherapy in the treatment of allergic fungal sinusitis. PMID- 9339799 TI - Comparison of sinus computed tomography staging systems. AB - In an attempt to establish a standardized rating system for CT of the paranasal sinuses, the Committee on Rhinology and Paranasal Sinus Disease of the American Academy of Otolaryngology-Head and Neck Surgery instituted a protocol for the review of sinus CT scans at six international sites. Fifty identical scans were rated by four otolaryngologists at each site according to five established sinus CT staging systems. Twenty of 24 reviewers repeated the rating session at least 1 week later to determine intrarater variability. The number of CT scans that could not be classified by a particular rating system ranged from 1.3% to 5.5%. The range of intrarater agreement (kappa = 0.39 to 0.74) exceeded that of interrater agreement (kappa = 0.18 to 0.49). A skewed distribution of CT scans resulted in a system with high rater agreement but poor ability to differentiate among disease states. The use of a numeric rating system to assign a score to each scan produced a comprehensive and disease-sensitive system, but one with low rater agreement. A precise definition of mucosal thickening in terms of millimeters appeared to enhance the raters' ability to assign stage and improve a system's comprehensiveness and reproducibility. On the basis of these findings, recommendations are made for the use of CT rating systems to study clinical outcomes in patients with chronic sinusitis. PMID- 9339800 TI - Outcomes research primer. PMID- 9339801 TI - The effects of an outpatient practice guideline at a teaching hospital: a prospective pilot study. AB - Practice guidelines (PGs) are becoming increasingly important in modern medicine. To study the effects of a PG, we performed a pilot study at a large, urban, public teaching hospital according to a prospective, observational research design with both concurrent and historic controls. Specifically, we studied the effects of a multidisciplinary PG for pediatric outpatient tonsillectomy and adenoidectomy on the process of health-care delivery. Variables in the health care process included patient compliance with clinic and surgery appointments, surgery time, operating room turnover, time in recovery room, unplanned admission rate, patient compliance with postoperative follow-up, provider compliance with guidelines, and hospital charges. Patients in the PG were found to have fewer preoperative laboratory tests, decreased duplication of services, and shorter operating room turnover times. Provider compliance with the PG varied by service and was intermittent at first but improved gradually. There was a trend toward improved compliance with postoperative follow-up in patients in the PG. Provider opinions concerning the guideline were positive. This pilot study demonstrates several advantages and disadvantages of the use of PGs in the outpatient setting and in a teaching hospital. PMID- 9339802 TI - Computerized dynamic platform posturography. AB - Computerized dynamic platform posturography is defined in this technology assessment. The review discusses what computerized dynamic platform posturography measures, what the reliability and validity of the information are, and the uniqueness of the information provided. The clinical contribution and indications for testing are discussed. There are comments on future directions for research on computerized dynamic platform posturography and a summary and conclusion. PMID- 9339803 TI - Phonatory characteristics of patients undergoing thyroidectomy without laryngeal nerve injury. AB - Complications that arise after thyroid surgery may be associated with infection, hemorrhage, hormonal problems, and laryngeal nerve injury. Voice alteration after thyroidectomy is usually caused by recurrent or superior laryngeal nerve injury. This voice dysfunction may also be associated with laryngotracheal fixation with impairment of vertical movement or by temporary malfunction of the strap muscles after surgery. In this study, we evaluated the voice function phonetically before and after thyroidectomy in 54 patients, although function of the recurrent and superior laryngeal nerves was normal. During surgery, the superior and recurrent laryngeal nerves were identified and protected, and after surgery electromyographic testing of the cricothyroid muscle was performed. Typical voice symptoms after surgery were easy fatigue during phonation and difficulty with high pitch and singing voice. Acoustic analysis revealed that the phonation time and fundamental frequency were not changed after surgery, but the speaking fundamental frequency, range of speaking fundamental frequency, and vocal range were significantly diminished after surgery. These data allowed us to suggest that the cause of voice dysfunction is not seen in neural lesions, but in a disturbance of the extralaryngeal skeleton. These voice changes emphasize the importance of the extralaryngeal mechanism for pitch control. PMID- 9339804 TI - Effects of topical otic preparations on hearing in chronic otitis media. AB - Most of the topical otic preparations have been shown to cause ototoxicity. In this study ciprofloxacin hydrochloride, a relatively new topical agent, and gentamicin sulfate were studied in two groups of 20 patients with chronic otitis media. Patients were randomly selected to receive either ciprofloxacin (200 microg/ml) or gentamicin sulfate (5 mg/ml) locally, five drops three times a day for 10 days. Clinical response was seen in 20 of 20 patients in the ciprofloxacin group compared with 6 of 20 patients in the gentamicin group. Audiometric evaluation revealed no significant ototoxic effect in either group. In fact, hearing thresholds were slightly better than pretreatment levels in both groups. PMID- 9339805 TI - Lymphangioma: an unusual cause of pharyngeal and epiglottic edema. PMID- 9339806 TI - Liposarcoma of the scalp: a case report and review of the literature. PMID- 9339807 TI - Pleomorphic adenoma of the larynx: a case and review of the literature in Japan. PMID- 9339808 TI - Rare otolaryngologic presentation of hydatid cyst. PMID- 9339809 TI - Recirculation of mucus via accessory ostia causing chronic maxillary sinus disease. PMID- 9339810 TI - Physician workforce in otolaryngology. PMID- 9339811 TI - Treatment of acute mastoiditis. PMID- 9339812 TI - Persistent decrease of the dopamine-synthesizing enzyme tyrosine hydroxylase in the rhesus monkey retina after chronic lead exposure. AB - One of the toxic effects of lead in the CNS is an altered functional state of the catecholamine system, especially a reduction in the activity of tyrosine hydroxylase (TH), the rate-limiting enzyme of catecholamine synthesis. Here we report on a lead-induced decrease in TH-content in neurones of the rhesus monkey retina. Rhesus monkeys were pre- and postnatally exposed to 0, 350, or 600 ppm of lead acetate (Pb) in the diet over 9 years. Lead exposure was followed by a 35 month period of lead-free diet. During this period, blood lead levels of the treated animals declined to nearly those of the untreated controls. Subsequently the animals were sacrificed and the retinas processed for TH immunocytochemistry. The fluorescent dye FITC was used to visualise the antibody reaction. Photometric measurements of the fluorescence intensity of stained neurones were made with a laser scanning microscope. In the rhesus monkey retina two types of TH immunoreactive neurones are present. In the bright fluorescent type, lead exposure resulted in decreased fluorescence intensity and altered the intensity profile of the TH-immunoreactive cells in a dose-dependent manner. In these cells, fluorescence intensity was 0.53 and 0.22 for 350 ppm Pb and 600 ppm Pb respectively when the fluorescence intensity of the untreated controls (0 ppm Pb) is taken as 1. Both lead doses also reduced the number of ascending fibres in the inner nuclear layer and the dense staining of fibres in sublayer 1 of the inner plexiform layer. The weakly fluorescent cell type disappeared to a large extent under 350 ppm Pb treatment and was not detectable in the 600 ppm Pb group. The results demonstrate that lead exposure affects the dopaminergic retinal amacrine cells by reducing the TH-content in these neurones and that this neurotoxic effect persists beyond the end of exposure. PMID- 9339813 TI - Altered expression of polysialylated NCAM in mouse hippocampus following trimethyltin administration. AB - Proper structuring of neural connections in the hippocampus is mediated by cell adhesion molecules, membrane-linked proteins involved in cell recognition and stabilization of cytoarchitecture. Modulated expression of the neural cell adhesion molecule (NCAM) at the synapse permits plasticity required for both learning and memory. Polysialylation of NCAM, particularly the synapse-specific 180 kDa isoform (NCAM180), allows hippocampal neurons to alter their neuronal connections during learning acquisition and memory consolidation in mature brain. These activity-dependent changes in NCAM expression represent a sensitive target for neurotoxicity. Trimethyltin (TMT), a potent hippocampal neurotoxicant, alters total NCAM expression in whole mouse hippocampus and impairs learning in rodents. To investigate the expression of polysialylated NCAM following TMT administration, Swiss-Webster mice were injected (i.p.) with 2.0 or 3.0 mg TMT/kg and sacrificed 6 hrs to 7 days later. Immunocytochemical staining for polysialylated NCAM (PSA-NCAM) revealed marked reduction of staining of hippocampal dentate granule cells 6-72 hours after TMT treatment. Partial recovery of hippocampal polysialylated NCAM was observed after 7 days. Immunoblot data indicated that loss of PSA-NCAM expression paralleled reductions seen in NCAM180 and markers of cytoskeletal integrity. Assays for proteolytic activity in hippocampus revealed rapid, reversible protease activation which correlated temporally with the reduction of NCAM180 and PSA-NCAM. Proteolytic degradation following hippocampal injury may serve to disrupt NCAM-mediated adhesion. Protracted loss of polysialylated NCAM in dentate gyrus following injury may serve as a useful marker in toxicant-induced learning disorders. PMID- 9339814 TI - Functional alterations and apoptotic cell death in the retina following developmental or adult lead exposure. AB - Long-term visual system deficits occur in man and animals following developmental and occupational lead exposure. Recent experimental data suggests that the adult brain is not altered following lead exposure. Therefore, the aim of these studies was to use the retina as a CNS model to examine and compare the morphological, biochemical and electroretinographic (ERG) changes occurring in rats exposed to low or moderate levels of lead during development (0-21 days of age) with those occurring in adult rats with equivalent blood and retinal levels of lead for three or six weeks. Five main results were obtained. First, developmental and adult lead exposure for six weeks produced age- and dose-dependent retinal degeneration such that rods and bipolar cells were selectively lost. At the ultrastructural level, all dying cells exhibited the classical morphological features of apoptotic cell death. Second, in the lead-exposed groups, the decrease in the number of rods was correlated with the loss of rhodopsin content per eye confirming that rods were directly affected by lead. Third, single-flash rod ERGs and cone ERGs obtained from developmentally and adult lead-exposed rats demonstrated that there were age- and dose-dependent decreases in the rod a-wave and b-wave sensitivity and maximum amplitudes without any effect on cones. In adult rats exposed to lead for three weeks, qualitatively similar ERG changes occurred in the absence of cell loss or decrease in rhodopsin content. Fourth, developmental and adult lead exposure for three and six weeks produced age- and dose-dependent decreases in retinal cGMP phosphodiesterase (PDE) activity resulting in increased cGMP levels. Fifth, picomolar to micromolar concentration of free lead directly inhibited rat retinal and purified bovine rod cGMP PDE. In summary, there are three main conclusions. First, the retinas of developing and adult rats exposed to lead exhibit qualitatively similar rod-mediated ERG alterations as well as rod and bipolar apoptotic cell death. Thus, developing and mature retinas are both sensitive to the adverse effects of lead: albeit to significantly different extents. Second, a similar biochemical mechanism such as the inhibition of rod and bipolar cell cGMP PDE, varying only in degree and duration, underlies both the lead-induced ERG rod-mediated deficits and the rod and bipolar apoptotic cell death. Third, the composite results from our experiments and those of others suggest that the developing retina might be more sensitive to preweaning lead exposure than the hippocampus and thus may serve as a good model for studying the cellular and molecular mechanisms underlying developmental and adult lead neurotoxicity. PMID- 9339815 TI - Glutamatergic system and developmental lead neurotoxicity. AB - The excitatory amino acid receptor subtype N-Methyl-D-Aspartate (NMDA) has been the focus of intense investigation during the last decade because of its role in brain development and synaptic plasticity and in pathological conditions associated with excitotoxicity. This receptor complex has recently been identified to be a target for lead (Pb2+) effects on the developing central nervous system. This presentation is an overview of studies demonstrating the interaction of Pb2+ with the NMDA receptor and its potential effects on cellular function and cognition. PMID- 9339816 TI - Relationships between Pb-induced changes in neurotransmitter system function and behavioral toxicity. AB - Lead (Pb) exposure impairs learning and results in changes in Fixed Interval (FI) schedule-controlled behavior in experimental animal studies. Studies from our laboratory suggest a major involvement of dopamine systems, and perhaps nucleus accumbens, in the FI performance changes. Dopaminergic (DA), but not other classes of compounds differentially alter FI response rates of control and Pb treated rats. Marked changes in nucleus accumbens but not striatal D2 and dopamine uptake sites occur in response to Pb. Further, the irreversible DA antagonist EEDQ microinjected into nucleus accumbens but not into striatum suppresses FI performance. In contrast, glutamatergic system disturbances appear to play a key role in the learning impairments caused by Pb. Glutamatergic (GLU) compounds differentially alter learning accuracy levels in control vs. Pb-treated rats, whereas DA compounds do not. Accuracy levels in the learning paradigm were positively correlated with numbers of MK-801 and glutamate binding sites in control rats; these correlations were reversed by Pb exposure. Future studies should be designed to confirm these relationships, to examine the brain regions involved, to determine the extent to which other behaviors known to be mediated by GLU/DA mesocorticolimbic systems are disrupted by Pb, and to address the significance of developmental period of exposure to these effects as well as their reversibility. PMID- 9339817 TI - Behavioral and neurochemical dynamics of neurotoxic meso-striatal dopamine lesions. AB - Here we review some of our recent experiments where the neurotoxin 6 hydroxydopamine had been used to unilaterally damage nigro-striatal dopamine neurons in the rat. In this lesion model we studied relationships between the degree of neostriatal dopamine loss, the resulting behavioral deficits, possible recovery therefrom, and mechanisms which may be related to such recovery. Special attention is given to animals with more moderate dopamine lesions, since these may be particularly suitable to study functional recovery. Our behavioral studies showed that initially after lesion (day 1), asymmetries of turning and scanning behavior were observed over a wide range of neostriatal DA depletions, whereas after one week these had recovered to symmetry except in animals with residual neostriatal dopamine levels of 20% or less. Subsequent experiments showed that behavioral asymmetries can also be induced in animals with less severe lesions (residual DA levels 45-65%), since ipsiversive asymmetries in scanning and turning were observed when such animals were challenged systemically with the mixed dopamine receptor agonist apomorphine. Finally, a weak ipsiversive asymmetry could also be induced in such animals by the more selective D2-agonist LY171555, whereas the D1-agonist SKF38393 was ineffective. These results are discussed with respect to the neural mechanisms determining deficits and recovery after such dopamine lesions and their relevance to similar clinical phenomena, namely Parkinson's disease. Here, special attention is given to the role of mechanisms, which can regulate compensatory increases of dopamine activity in those neurons which have survived the lesion. PMID- 9339818 TI - Cellular aspects of persistent neurotoxicants: effects of Pb2+ on neuronal nicotinic acetylcholine receptors. AB - Among persistent neurotoxicants inorganic lead (Pb2+) is known to cause a variety of cellular effects associated with alterations in neuronal performance. Here we describe differential effects of Pb2+ on distinct subtypes of neuronal nicotinic acetylcholine receptors (nAChR) in Xenopus oocytes expressing various combinations of alpha and beta receptor subunits. Transient inward currents evoked by ACh in voltage clamped oocytes expressing alpha4beta2 and alpha3beta4 nAChR are inhibited by external Pb2+ at concentrations in the range of 1 nM-200 microM. The sensitivities of oocytes expressing alpha4beta2 and alpha3beta4 nAChR vary greatly. Additional experiments suggest that this variation may be due to receptor heterogeneity, which is demonstrated for alpha4beta2 nAChR in cell attached patch clamp experiments. Conversely, the inward current evoked by ACh in oocytes expressing alpha3beta2 nAChR is potentiated by Pb2+ at concentrations in the range of 1-250 microM. The potentiating effect shows little variation between oocytes and is similar for inward currents evoked by low and high concentrations of ACh. The distinct effects on alpha3beta2 nAChR as compared to alpha4beta2 and alpha3beta4 nAChR suggest that Pb2+ interacts with both alpha and beta subunits of the neuronal nAChR. The effects of Pb2+ on defined subtypes of nAChR in oocytes are compared to those on ligand-gated ion channels in intact cells. PMID- 9339819 TI - Developmental neurotoxicity of environmental agents in the neonate. AB - The development of an organism includes periods that can be critical for its normal maturation. One such appears to occur during perinatal development of the brain, the so-called 'brain growth spurt'. This period in the development of the mammalian brain is associated with numerous biochemical changes that transform the feto-neonatal brain into that of the mature adult. We have observed that low dose exposure to environmental agents such as DDT, pyrethroids, organophosphates, nicotine, paraquat and polychlorinated biphenyls (PCBs) during the 'brain growth spurt' can lead to irreversible changes in adult brain function in the mouse. The induction of behavioural and cholinergic disturbances in the adult animal appears to be limited to a short period during neonatal development, around postnatal day 10, and following doses that apparently have no permanent effects when administered to the adult animal. Furthermore, neonatal exposure to a low dose of a neurotoxic agent can lead to an increased susceptibility in adults to an agent having a similar neurotoxic action, resulting in additional behavioural disturbances and learning disabilities. PMID- 9339820 TI - Neurochemical effects of polychlorinated biphenyls: an overview and identification of research needs. AB - The PCBs are members of the halogenated hydrocarbon class of environmental chemicals that includes the dibenzofurans and dioxins. The PCBs were used over a period of 40 years for number of industrial purposes. Their appearance in the ecosystem and biological samples from wildlife, as well as documented cases of accidental poisoning led to the banning of their manufacture in 1977. The PCBs continue to be of concern to environmental toxicologists because of their persistence in the environment and reports that exposure to relatively low levels may be associated with subtle behavioral and neurological deficits, particularly if exposure occurs during development. Developmental neurotoxicity of PCBs has been reported in humans and confirmed in several laboratory animal species, including non-human primates. During the last 20 years, there has been an attempt to understand the cellular bases of PCB-induced behavioral and neurological effects in animal models. Exposure of adult animals to a single, relatively high dose of PCBs decreases the content of several brain neurotransmitters, while repeated exposure to lower PCB doses appears to affect brain DA metabolism. The mechanism by which PCB affects DA remains unclear. It is now known that some PCB congeners have a structural configuration that facilitates binding to an aryl hydrocarbon (Ah) receptor like other polychlorinated compounds, including 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Some PCB congeners, on the other hand, have structural characteristics, e.g., non-coplanarity, that diminish access to the Ah receptor. Non-TCDD-like PCB congeners that appear in the brain following in vivo exposure demonstrate the highest potency in terms of decreasing DA content in PC 12 cells and inhibiting calcium homeostasis mechanisms in vitro. The biological significance of the effects of the PCBs on DA content or calcium homeostasis with regard to the behavioral and neurological effects observed following developmental exposure in vivo is not clear. Recent research, however, suggests that PCBs can alter a number of physiological processes that may be important for development. For example, PCB-induced alterations in thyroid function during development may underlie some of the developmental effects of PCBs reported in humans and animal models. Additional research on the PCBs seems warranted in a number of areas, including the: 1) structural requirements necessary for binding to the Ah-receptor, 2) mechanism(s) of PCB-induced alterations in DA content and calcium homeostasis in vitro, 3) relationship between observed neurochemical effects in vitro and effects in vivo, and 4) relationship between PCB-induced neurochemical effects and crucial developmental processes such as those controlled by thyroid hormone development. PMID- 9339821 TI - Interactions between steroid hormones and the nervous system. AB - After a short description of the endocrine and nervous system and their similarities and differences, interactions between both parts are mainly dealt with in this brief review. This is especially exemplified by the modulation of receptors for neurotransmitters, of ion channels and other membrane components by neurosteroids and steroid hormones and the physiological significance of this regulation. Both classes of hormones can act via nongenomic and genomic actions. As biological responses to neurosteroids and steroid hormones in the nervous system exocytosis of peptide hormones and neurosteroids, regulation of neurotransmitter release, sexual brain development and male behavior and cell death are described. Neurotransmitters and their receptors are also modulated in their expression and biological activity by steroid hormones and neurosteroids in non-neuronal tissues, as demonstrated for GABA-receptors in the uterus and the embryonic cholinergic system. Because of the close links between endocrine and nervous system agents toxic for one component may also interfere with the other one. PMID- 9339822 TI - Actions of pyrethroid insecticides on voltage-gated chloride channels in neuroblastoma cells. AB - Pyrethroid insecticides have the potential to act at several sites in excitable tissues in addition to their primary effect on sodium channels. We have assessed one such site, the voltage gated chloride channel. Following our previous demonstration of actions of deltamethrin on the voltage-gated chloride channel in rats and in neuroblastoma cells, we examined the dose-response relationship for deltamethrin by patch clamp analysis of voltage-gated channels in partially differentiated NIE115 cells. Open channel probability (P0) was assessed in channels stepped from 0 to +20 mV for 10 seconds. Deltamethrin significantly decreased P0 by 0.237+/-0.070 at 10(-10) M and, although construction of a dose response relationship was rendered difficult by the very low water solubility of the pyrethroids, a decrease of 0.968+/-0.140 was reached at the highest concentration tested, 10(-4) M. A second Type II pyrethroid cypermethrin also decreased P0, by 0.430+/-0.09 at 5 x 10(-6) M, but the Type I pyrethroid cismethrin produced a decrease of only 0.188+/-0.08 at 2 x 10(-5) M. These effects were seen in 340 pS conductance, calcium-independent channels. A sample of 9 calcium-dependent channels with a conductance of 225 pS failed to show any response to deltamethrin. We conclude that actions on calcium-independent voltage gated chloride channels are likely to contribute significantly to type II pyrethroid toxicity. PMID- 9339823 TI - Botulinum toxin: from poison to remedy. AB - Botulinum toxins, exotoxins of Clostridium botulinum, are the most toxic naturally occurring substances known to man. For more than a century they are known to be the cause of botulism, a nowadays rare intoxication with spoiled food that leads to generalized flaccid weakness of striated muscle including pharyngeal and respiratory musculature. The toxins act primarily at peripheral cholinergic motor nerve endings by blocking the release of the neurotransmitter acetylcholine. As a consequence, action potentials in the motor nerve can no longer be transmitted to the muscle. This lack in transmission, clinically appearing as weakness, may disable or actually critically endanger affected patients. However, in certain neurological diseases characterized by an abnormal increase in muscle tone or activity, for example dystonia or spasticity, a reduction in signal transmission may actually be beneficial. Around 1980 local injections of minute amounts (in the order of 0.5 ng) of Botulinum toxin type A were first successfully used in a neurological disorder named blepharospasm which is characterized by an involuntary squinting of the eyes. Since then Botulinum toxin has developed rapidly from a frightful poison to a safe therapeutic agent with a remarkable beneficial impact on the quality of life of many thousands of patients worldwide. This review tries to outline in brief the characteristics of Botulinum toxins, their mechanism of action and the various indications for clinical use as a therapeutic agent. PMID- 9339824 TI - Behavioral and neurochemical effects of acute and repeated administration of triadimefon in the male rat. AB - The effects of triadimefon (TDF) were examined in male Sprague-Dawley rats. In this study, the acute administration of TDF (100 mg/kg) was found to significantly increase locomotor activity and induce stereotyped behavior. Acute administration of TDF was also found to significantly increase dopamine (DA) and homovanillic acid (HVA) levels while the dihydroxyphenylacetic acid (DOPAC) level remained unchanged in both the nucleus accumbens (NA) and striatal (ST) tissues when compared to control. Furthermore, DOPAC:DA ratios were significantly reduced in both brain regions suggesting an increase in DA turn overrate. On the other hand, in animals receiving repeated TDF administration, only the HVA level was significantly increased in both the ST and NA. TDF neither competed for binding to D2, D3 or D4 DA receptors nor altered the Kd or the Bmax of [3H] SCH 23390 and [3H] spiperone recognition sites associated with striatal D1 and D2 receptors, respectively. Meanwhile, TDF competed with [3H] GBR 12935 for binding to DA transporter sites with strong affinity, but repeated treatment with TDF had no sustained or cumulative effect on the DA transporter system. These results clearly show that acute TDF-induced behavioral effects may not be via binding to DA receptors, but through the interaction with DA transporter binding sites. Also, TDF does not appear to produce cumulative effects in the parameters evaluated. PMID- 9339825 TI - Acute energy deprivation syndromes: investigation of m-dinitrobenzene and alpha chlorohydrin toxicity on immortalized rat brain microvessel endothelial cells. AB - Acute energy deprivation syndromes share a common pattern of CNS pathology resulting in symmetrical spongiform brain stem lesions in rodents. However, some toxicants are proposed to act on astrocytes alone (alpha-chlorohydrin) whilst others are associated with petechial haemorrhages and blood-brain barrier (BBB) breakdown (m-dinitrobenzene). In this study, we investigated the toxicity of alpha-chlorohydrin (alpha-CH) and m-dinitrobenzene (m-DNB) in an in vitro BBB model, the rat brain capillary endothelial cell line RBE4. Cytotoxicity was observed after treatment of RBE4 cells with both toxicants, as manifested by a decrease in protein content and MTT reduction (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide) over control values at concentrations > or = 1 mM for m-DNB and > or = 20 mM for alpha-CH. m-DNB caused a dose-dependent increase in glucose consumption and lactate production in RBE4 cells, while alpha-CH had no effect on these parameters. The distribution of F-actin at the cell margin observed in control cultures was changed to a diffuse pattern over the cell cytoplasm after treatment with both toxins at subcytotoxic concentrations. However, a reduction in F-actin content was only observed at concentrations > or = 1 mM for m-DNB and > or = 20 mM for alpha-CH. The permeability of RBE4 cell monolayers cultured on filters above primary rat astrocytes was measured using 14C-sucrose (M.Wt.=342) and FITC-dextran (M.Wt.=4000). m-DNB (0.5 mM) increased the permeability of RBE4 cell monolayers to both tracers, while alpha-CH (30 mM) had no effect. The results from this study indicate that m-DNB may have direct toxic effects on brain endothelial cells which lead to loss of barrier function. Whilst alpha-CH caused some toxic effects in RBE4 cells, it did not alter endothelial cell monolayer permeability. PMID- 9339826 TI - Cumulative blood lead levels and neurobehavioral test performance. AB - The current scientific literature provides inadequate evidence to conclude whether or not cumulative exposure to or absorption of lead adversely affects performance in neurobehavioral tests in adults. One of reasons for this controversy is the lack of studies with good cumulative exposure to or dose of lead. The aims of this study are to compare the neurobehavioral test performances of a group of lead-exposed workers and a referent group, and to study the association of the neurobehavioral test performances with concurrent blood lead levels and cumulative blood lead levels. Fifty lead battery workers and 97 non exposed (referent) workers from a vehicle maintenance workshop were evaluated on their neurobehavioral performance using the World Health Organization Neurobehavioral Core Test Battery (WHO-NCTB). The geometric mean concurrent blood lead (ConPb) of the exposed and referent groups were 37.1 (range 13.2-64.6) microg/100 ml and 6.1 (range 2.4-12.4) microg/100 ml, respectively. Cumulative blood lead (CumPb) was defined as area under the curve for the number of years each worker was exposed to lead (three workers previous blood lead results were not available). ConPb and CumPb were used to study the association with the neurobehavioral test results. The exposed group had significantly poorer manual dexterity, perceptual-motor speed, and motor steadiness compared with the referents. The standardized partial regression coefficients were higher for CumPb than ConPb for most of the neurobehavioral test results. In the group >35 years old, there were significantly stronger associations between CumPb and Digit Symbol and Trail Making Part A results than for ConPb which are tests of perceptual and motor skills. CumPb was a better predictor than ConPb of the effects of lead on neurobehavioral performances. PMID- 9339827 TI - Prenatal and postnatal lead exposure induces 70 kDa heat shock protein in young rat brain prior to changes in astrocyte cytoskeleton. AB - In previous studies we found that chronic postnatal (PN) lead exposure [1 g% (w/v)] induced astroglial hypertrophy in rat hippocampus. Since astrocytic responses change upon the stage during which exposure occurs, astroglial reactions in cerebral cortex and hippocampus of young animals were studied and compared when exposure began during development. Lead-treatment started 90 days prior to mating, and was maintained during gestation and after birth up to PN160. Alterations observed from PN21 to PN140 were assessed by antibodies to the 70kDa heat shock proteins (hsp), glial fibrillary acidic protein (GFAP) and vimentin (VIM) using immunohistochemistry, transmission electron microscopy (TEM), and computer assisted image analysis. The induction of hsp was seen from PN21 to PN45 in non-pyramidal neurons and astrocytes, and at the same time, astroglial swelling was noticed. After PN45 the resolution of this edema coincided with an increase of gliofilaments and GFAP and VIM immunoreaction (PN60-PN90). Recovery of VIM expression persisted after PN120 in the hilus; meanwhile, lipofuscin-like bodies appeared in neurons and astrocytes. Lead exposure during rapid brain growth induced hsp after weaning in neurons and astrocytes prior to astrocyte cytoskeletal changes. Astroglial and neuronal alterations could modify complex neuron-glia interactions, disturbing brain function in consequence. PMID- 9339828 TI - Effects of prenatal methylmercury exposure from a high fish diet on developmental milestones in the Seychelles Child Development Study. AB - Mercury is widespread in the environment and exists in several physical and chemical forms. Prenatal exposure to methylmercury disrupts brain development. The most common mode of prenatal methylmercury exposure is maternal fish consumption. Studies of human prenatal exposure in Iraq following maternal ingestion of methylmercury treated grain suggested that maternal hair mercury concentrations above 10 ppm may be related to delayed developmental milestones and neurological abnormalities. This level of exposure can be achieved by frequent consumption of fish. The Seychelles Child Development Study analyzed developmental milestones similar to those determined in Iraq in a large controlled, prospective study of children exposed prenatally to methylmercury when their mothers ate fish. As part of this ongoing study, cohort children were evaluated at 6.5, 19, 29, and 66 months of age. At 19 months care-givers were asked at what age the child walked (n=720 out of 738) and talked (n=680). Prenatal mercury exposure was determined by atomic absorption analysis of maternal hair segments corresponding to hair growth during the pregnancy. The median mercury level in maternal hair was 5.8 ppm with a range of 0.5-26.7 ppm. The mean age (in months) at walking was 10.7 (SD = 1.9) for females and 10.6 (SD = 2.0) for males. The mean age at talking (in months) was 10.5 (SD = 2.6) for females, and 11.0 (SD = 2.9) for males. After adjusting for covariates and statistical outliers, no association was found between the age at which Seychellois children walked or talked and prenatal exposure to mercury. Normal ages at achievement of the developmental milestones walking and talking were found in Seychellois toddlers following prenatal exposure to methylmercury from a maternal fish diet. These results do not support the lowest effect levels in young children following prenatal methylmercury exposure predicted by the dose response analysis of the Iraq data. More detailed studies in older children are needed to determine if there are adverse effects in fish eating populations. PMID- 9339829 TI - Effects of the naturally occurring food mycotoxin ochratoxin A on brain cells in culture. AB - The potential of ochratoxin A (OTA) to damage brain cells was studied by using a three-dimensional cell culture system as model for the developing brain. Aggregating cell cultures of foetal rat telencephalon were tested either during an early developmental period, or during a phase of advanced maturation, over a wide range of OTA concentrations (0.4 nM to 50 microM). By monitoring changes in activities of cell type-specific enzymes (ChAt and GAD, for cholinergic and GABAergic neurones, respectively, GS for astrocytes and CNP for oligodendrocytes), the concentration-dependent toxicity and neurodevelopmental effects of OTA were determined. OTA proved to be highly toxic, since a 10-day treatment at 50 nM caused a general cytotoxicity in both mature and immature cultures. At 10 nM of OTA, cell type-specific effects were observed: in immature cultures, a loss in neuronal and oligodendroglial enzyme activities, and an increase in the activity of the astroglial marker glutamine synthetase were found, Furthermore, at 2 and 10 nM of OTA, a clustering of microglial cells was observed. In mature cultures, OTA was somewhat less potent, but caused a similar pattern of toxic effects. A 24 h-treatment with OTA resulted in a concentration dependent decrease in protein synthesis, with IC50 values of 25 nM and 33 nM for immature and mature cultures respectively. Acute (24 h) treatment at high OTA concentrations (10 to 50 microM) caused a significant increase in reactive oxygen species formation, as measured by the intracellular oxidation of 2',7' dichlorofluorescin. These results suggest that OTA has the potential to be a potent toxicant to brain cells, and that its effects at nanomolar concentrations are primarily due to the inhibition of protein synthesis, whereas ROS seem not to be involved in the toxicity mediated by a chronic exposure to OTA at such low concentrations. PMID- 9339830 TI - Methylmercury decreases IL-1beta immunoreactivity in the nervous system of the developing frog Xenopus laevis. AB - In aquatic ecosystems, mercury can become methylated and act as a potent environmental toxin, producing developmental and neurotoxic effects in a variety of species, including frogs. Molecular indicators provide a means of assessing exposure to methylmercury and for understanding how it and other environmental toxins alter cellular function. Molecules such as growth or survival factors, and cytokines are good candidates for molecular indicators of exposure and/or damage because they are intimately related to cell and molecular processes that underlie normal growth and function. The cytokine, IL-1beta, was measured in whole frog embryos using Western blot methods and in specific structures using immunocytochemistry after exposure to 0, 10, 25, 50 and 100 parts per billion (ppb) methylmercury chloride (mmc). We observed no significant changes in total IL-1beta in whole embryo extracts. However, statistically significant decreases in IL-1beta were observed in the Vth cranial ganglion and myotomal blocks of Xenopus laevis embryos exposed to concentrations greater than or equal to 50 ppb mmc. In addition, increased mortality and alterations in gross morphology and behavior were altered by these same concentrations of mmc. Thus, frog embryos are highly susceptible to low levels of mmc contamination, and IL-1beta is an indicator of mmc exposure in the nervous system. PMID- 9339831 TI - GK 11: promising additional neuroprotective therapy for organophosphate poisoning. AB - Recent experiments with primates have demonstrated that treatment with atropine/pralidoxime/diazepam, even if administered immediately after organophosphate exposure, does not totally prevent neuronal brain damage. Using primates, we have studied, for the first time, the ability of GK-11 (gacyclidine), an antiglutamatergic drug in the process of agreement for human use, given as an additional therapy, to counteract the neuropathology due to organophosphate exposure that persists after classical treatment with oxime/atropine/benzodiazepine. We have also examined the recovery of the organophosphate-intoxicated primates. Male Cynomolgus monkeys were pretreated 1 hour before poisoning with pyridostigmine, then intoxicated with 8 LD50 of soman and immediately treated with the combination pralidoxime/atropine/diazepam. Some of the animals also received GK-11 at 0.01; 0.03 or 0.1 mg/kg (i.v.) 10 minutes after soman challenge. Recovery of the primates (reflexes, movements, feeding) and the neuropathological changes that occurred three weeks after intoxication (histological examinations and neuronal cell density measurement) were compared in GK-11-treated and control animals. At all doses tested, GK-11 prevented the neuronal rarefaction of the frontoparietal cortex that was observed in soman intoxicated animals that received only oxime/atropine/diazepam. Moreover, the 0.01 mg/kg dose of GK-11 improved the early recovery of intoxicated primates from 1 day after intoxication. In the view of the most effective management of organophosphate intoxication that is currently available, GK-11 thus appears to be a promising additional neuroprotective therapy. This drug is presently being evaluated in a human clinical trial for a different neuroprotective indication. PMID- 9339832 TI - Inhibition of L-aromatic amino acid decarboxylase by polychlorinated biphenyls. AB - PC12 cells were used to examine the mechanisms by which polychlorinated biphenyls (PCBs) reduce cellular levels of dopamine (DA). In cells treated 3 days with Aroclor 1254, 2,2',5,5'-tetrachlorobiphenyl (2,2',5,5'-TCB), or 2,2',3,3',4,4' hexachlorobiphenyl (2,2',3,3',4,4'-HCB), the PCB-mediated reduction in 3H tyrosine uptake was observed only at high PCB concentrations that produced a reduction in DNA levels. The PCB congener, 2,2',4,4',5,5'-hexachlorobiphenyl (2,2',4,4',5,5'-HCB) did not produce a reduction in 3H-tyrosine uptake at any concentration tested. Thus, there were PCB concentrations at which a reduction in DA levels did not coincide with a decrease in 3H-tyrosine uptake, suggesting that inhibition of tyrosine uptake was not the primary mechanism by which PCBs reduce DA levels. Aroclor 1254-treated cells also exhibited elevated levels of DOPA, further supporting the conclusion that tyrosine levels were not limiting. Incubation of Aroclor 1254-pretreated cells with 3H-tyrosine resulted in a dose dependent increase in cellular levels of 3H-DOPA and decrease in cellular levels of 3H-DA, suggesting a PCB-mediated inhibition of the conversion of 3H-DOPA to 3H DA. When the media was supplemented with DOPA, Aroclor 1254-treated cells still exhibited reduced levels of DA, compared to control cells, even though the control and PCB-treated cells had similar cellular levels of DOPA. Thus, one mechanism by which PCBs may reduce cellular levels of DA is by inhibiting L aromatic amino acid decarboxylase-mediated conversion of DOPA to DA. The PCB congeners, 2,2',4,4'-TCB, 2,2',5,5'-TCB, and 2,2',4,4',5,5'-HCB, also produced dose-dependent increases in DOPA levels. The congener 2,2',3,3',4,4'-HCB did not produce an increase in DOPA levels, although it did mediate reductions in cellular DA levels. However, when PC12 cells were supplemented with DOPA, all four PCB congeners produced a similar reduction in DA levels, suggesting that the conversion of DOPA to DA was inhibited by the PCBs. PMID- 9339833 TI - Implication of external Mg2+ ions in the depolarization of smooth muscle cell membrane of the human chorionic placental vessels: an hypothesis. AB - The membrane potential (MP) of the smooth muscle cells of human allantochorial placental vessels is the major factor which explains the excitation-contraction coupling. Indeed, the smooth muscle cells of these vessels may be considered as tonic because they only respond to excitatory stimuli with graded depolarization. On the contrary, the phasic smooth muscles generate action potentials and some generate electrical slow waves as well. The depolarization of smooth muscle cell membranes may be a consequence of various factors, particularly the secretion of endothelium-derived depolarizing factors (EDDF). The modifications of the ionic composition of external medium interfere with the membrane potential values. The increase of [Mg2+]o (Cl- or SO4[2-]) depolarizes the membrane. This effect in vitro seems inconsistent with the known vasodilator action of external Mg2+. To explain this result, an hypothetical scheme is proposed on the probable targets of local and systemic Mg2+ effects: a direct action of external Mg2+ ions on the membrane potential by secretion of depolarizing endothelial factors which depolarize the membrane; an indirect action on the membrane potential by regulation of K+ and Ca2+ channels; an action on the concentration of internal Mg2+; an interaction with internal Ca2+ concentration and regulation of the Na+/Ca2+ exchanger. A data review of Mg2+ ions effects on smooth muscles is realized to corroborate this scheme. PMID- 9339834 TI - Ex vivo study of the effects of dietary magnesium deficiency on the formation and release of NO from endothelial cells and the sensitivity to NO of smooth muscle cells of thoracic aortas isolated from rats. AB - We found that the contractile responses to phenylephrine (PE) of isolated endothelium-intact thoracic aortas were enhanced by a nitric oxide (NO) synthase inhibitor, nitro-L-arginine (LNAG). and the magnitude of this enhancement was significantly greater in Mg-deficient than control rats. In this study, we used a sandwich method to evaluate the effects of dietary Mg deficiency on the ability of vascular endothelial cells to form and/or release NO and on the sensitivity to NO of vascular smooth muscle cells. Male Wistar rats were fed a Mg-deficient (10 mg Mg/kg diet) or control (700 mg Mg/kg diet) diet for 30 days. With the sandwich method, when endothelium-denuded thoracic aortas from chow fed rats were used as the assay vessels, and endothelium-intact aortas from control or Mg-deficient rats were used as the NO-donor vessels, neither the aortic contractile responses to PE nor the enhancement by LNAG of these responses in control and Mg-deficient rats differed significantly. When endothelium-denuded thoracic aortas from control or Mg deficient rats were used as the assay vessels and endothelium intact aortas from chow fed rats were used as NO-donor vessels, LNAG enhanced PE induced contractions to a significantly greater extent in Mg-deficient than control rats. Furthermore, sodium nitroprusside (a NO donor) had a greater relaxant effect on endothelium-denuded thoracic aortas isolated from Mg-deficient than control rats. These ex vivo results suggest that dietary Mg deficiency in rats has little effect on the formation and/or release of NO by aortic endothelial cells. but it does appear to increase the sensitivity to NO of aortic smooth muscle cells. PMID- 9339835 TI - Ex vivo study of the increased sensitivity to NO of endothelium-denuded thoracic aortas isolated from dietary magnesium-deficient rats. AB - The mechanisms underlying the enhanced relaxant responses to sodium nitroprusside (SNP) associated with magnesium (Mg) deficiency were examined using endothelium denuded thoracic aortas isolated from rats with dietary Mg deficiency. This enhancement of SNP-induced relaxation was abolished or depressed in the presence of methylene blue (a guanylate cyclase inhibitor). The relaxant responses to 8 bromo cyclic GMP (8-Br cGMP; a membrane-permeable cGMP) of thoracic aortas isolated from Mg-deficient rats were enhanced like those to SNP. These enhanced relaxant responses were depressed by tetraethylammonium (a non-specific K+ channel blocker). Charybdotoxin, a large conductance Ca2+-activated K+ (K[Ca]) channel blocker, inhibited 8-Br cGMP-induced relaxations of aortas from Mg deficient and control rats. Apamin, a small conductance K(Ca) channel blocker, inhibited 8-Br cGMP-induced relaxations of aortas from Mg-deficient, but not control rats. The relaxant responses to cromakalim (an ATP-sensitive K+ channel opener) of the two groups were not significantly different. These ex vivo results show that dietary Mg deficiency in rats leads to increased sensitivity to NO of endothelium-denuded thoracic aortas in vitro and K(Ca) channel activation via cGMP may be involved in this enhancement. PMID- 9339836 TI - The activation by magnesium treatment of anti-oxidants eliminating the oxygen free radicals in Drosophila melanogaster in vivo. AB - Drosophila melanogaster adults of the Oregon R and Canton S wild-type stocks were fed for 24 h with 0.10 M, 0.25 M and 0.50 M MgCl2 in 1 per cent sugar solution. This treatment resulted in a significant increase (10-50 per cent) in the activities of the enzymes superoxide dismutase (SOD) and catalase as well as in the concentration of the non-enzymatic antioxidant glutathione (GSH). The increased activities could be due to the increased synthesis caused by the Mg treatment. PMID- 9339837 TI - Effects of low calcium and magnesium dietary intake on the central nervous system tissues of rats and calcium-magnesium related disorders in the amyotrophic lateral sclerosis focus in the Kii Peninsula of Japan. AB - Current epidemiological investigations in the Western Pacific including the Kii Peninsula of Japan, have suggested that environmental factors contribute to the pathogenetic process of amyotrophic lateral sclerosis (ALS) and parkinsonism dementia (PD). The condition of unbalanced minerals (a low content of calcium and magnesium, and a high content of aluminum) found in soil and drinking water in all three ALS foci was experimentally mimicked in our studies using rats. In rat groups maintained on unbalanced mineral diets, the calcium and magnesium contents of bones were lower than those fed a standard diet. In addition, the calcium content of CNS tissues showed higher values in the unbalanced diet groups (especially in the spinal cord of the low calcium and magnesium plus high aluminum diet group) than those in the standard diet group. The calcium content of other soft tissues as well as the CNS of rats fed unbalanced mineral diets was also higher than those on the standard diet. The magnesium content of soft tissues and spinal cord of rats was markedly lower in the low calcium and magnesium plus high aluminum diet group than in the other groups. Examination of tissues from six Kii Peninsula patients with ALS showed an average magnesium concentration in 26 CNS regions (cortical gray matter, white matter, basal ganglia, brain stem, spinal cord) significantly lower than that for five neurologically normal controls. The average calcium concentration in gray matter of ALS cases was significantly higher than that of controls. Interestingly, only 120 cases of calcification of spinal ligaments have been reported worldwide, and of these, 26 of 28 cases of calcification of spinal in the Kii Peninsula have been found to overlap the same geographic focal region as ALS. We analyzed the magnesium content of seven spinal vertebrae and 10 spinal ligaments of patients with calcification of spinal ligaments and the calcium content of five spinal bones compared with controls. The calcification of spinal ligaments patients had lower values for magnesium contents of bones and ligaments compared to controls and the calcium content of bones in these patients was significantly lower than that of controls. These data suggest that low dietary intake of calcium and magnesium over an extended period of time may contribute to the pathogenesis of patients with ALS and calcification of spinal ligaments. PMID- 9339838 TI - Magnesium and calcium in drinking water and cerebrovascular mortality in Taiwan. AB - The relationship between death from cerebrovascular disease and the levels of magnesium and calcium in drinking water was examined using an ecological design. The study area consisted of 227 municipalities in Taiwan. Data on the levels of magnesium and calcium in drinking water have been collected from the Taiwan Water Supply Corporation (TWSC). These levels of magnesium and calcium were compared using the standardized mortality ratios (SMRs) for cerebrovascular disease (1981 1990). A statistically significant inverse relationship was present between cerebrovascular mortality and levels of both magnesium and calcium after adjusting for urbanization index. After adjustment for calcium levels in drinking water and urbanization index, the weighted multivariate-adjusted regression coefficient indicated a decrease of 0.248 in the standardized mortality ratios (SMRs) for every 100 mg/L increase in magnesium levels in drinking water. The results from this study strengthen the hypothesis that magnesium in drinking water helps to prevent death from cerebrovascular disease. PMID- 9339839 TI - Parenteral magnesium loading test in the assessment of magnesium status in healthy adult French subjects. AB - Parenteral magnesium loading test has been proposed as an adequate mean to evaluate magnesium status. However, applied tests vary among different laboratories and standardized procedure is not available. In the present study, we assessed magnesium status in 32 healthy adult French subjects by magnesium loading test using MgCl2 and determination of magnesium concentrations in plasma and erythrocytes. We observed a positive correlation between plasma and erythrocyte magnesium concentrations, but there was no correlation between magnesium retention and basal urinary excretion of magnesium, and plasma or erythrocyte magnesium concentrations. PMID- 9339840 TI - Effect of an aerobic training on magnesium, trace elements and antioxidant systems in a Down syndrome population. AB - The aim of the present study was to determine the effect of an aerobic training on plasma and red blood cells' levels of magnesium, copper, selenium and zinc and on some oxidative stress parameters in a Down syndrome (DS) sample population. Sixteen young male adults with DS participated in the protocol. Among them, eight were randomly assigned to the control group and the remaining eight participated in a 16 week training programme consisting of 10 min warm-up followed by an aerobic session at a work intensity of 60 to 75 per cent of VO2 peak lasting from 15 to 25 min, increasing 5 min every 5 weeks and by a 5 min cool-down period, 3 days/week. Blood was withdrawn by butterfly from antecubital vein of each subject at fast, 2 days before the beginning of the programme and 2 days after its ending. Before the training period, when comparing the two groups, no significant differences were observed in the evaluated parameters. However, when comparing with a healthy population, red blood cells magnesium and plasma and red blood cells selenium mean values were low in both groups and mean SOD activity was 1.4 times higher. After the protocol the mean values of the minerals studied did not show significant differences between groups except for plasma zinc that was lower (p = 0.029) in the trained group. Plasma TBARS increase was significant in the trained group (p = 0.034) but not in the control group and plasma GSH of the trained group had a significantly higher increase than the control group (p = 0.003). The levels of plasma TBARS after the training programme that were inversely correlated with red blood cells GSSG levels (p = 0.023) and the higher increase of plasma GSH mean values observed, may be explained by the effect of the exercise period on the peroxidation and reduction of glutathione and also on the synthesis and efflux of GSH. Red blood cells magnesium levels remained low after the training programme which is in accordance with other studies. Plasma zinc decreased during the programme could be related to the activated expression of antioxidant mechanisms after the training. PMID- 9339841 TI - Chimpanzee pathogenic strains of HIV type 1. PMID- 9339842 TI - HIV type 1 strains pathogenic for chimpanzees. PMID- 9339843 TI - IgA immunity in HIV type 1-infected chimpanzees. I. Systemic immunity. AB - HIV infection in humans causes various aberrancies in both the cellular and humoral immune systems, including functional abnormalities of B lymphocytes. In many instances, dysfunction occurs in the regulation of serum IgA, resulting in elevated concentrations of this immunoglobulin isotype. To determine whether HIV 1-infected chimpanzees develop IgA abnormalities similar to those observed in humans, we quantified total IgA, IgG, and IgM levels in sera collected longitudinally from six HIV-infected chimpanzees and one uninfected control animal. In comparison to immunoglobulin levels in the uninfected animal, two of the six infected chimpanzees exhibited increases in serum immunoglobulins following infection with HIV. Two other infected animals showed a marked decrease in the three isotypes within 10 months of exposure to HIV, followed by a return to baseline levels. The remaining two HIV-infected chimpanzees displayed serum immunoglobulin levels that paralleled the baseline levels and did not show great deviation over a period of 20 to 45 months postinfection. ELISA analyses of the IgA subclasses revealed possible abnormalities of the IgA2 subclass within the two animals that did not display irregular IgA, IgG, or IgM responses to HIV-1. Specific IgG, IgA, IgA1, and IgA2 antibodies to HIV antigens were detected by an enzyme immunoassay (EIA) kit and by Western blot analysis with IgA, IgA1, and IgA2 antibodies directed against the env, gag, and pol gene products. Because IgG can mask the detection of HIV-specific IgA antibodies in infected humans, Western blots and EIAs were also performed on IgG-depleted chimpanzee sera. The results demonstrated that in some instances, IgA reactivity against HIV antigens can be enhanced on removal of IgG. This study indicates that HIV-1 is capable of inducing abnormalities in serum IgA expression in chimpanzees. These results might further understanding of how HIV affects humoral responses in infected humans. PMID- 9339844 TI - IgA immunity in HIV type 1-infected chimpanzees. II. Mucosal immunity. AB - Vaginal wash fluids from chimpanzees cervically infected with HIV-1 and saliva from intravenously and cervically infected chimpanzees were analyzed for total IgA, IgA1, IgA2, IgG, and albumin concentrations and for reactivity against HIV 1. No overt abnormalities were detected in salivary immunoglobulin or albumin concentrations in either group of animals. Anti-HIV IgA and IgA subclass antibodies were demonstrated in saliva from five of six intravenously infected chimpanzees and in two of four cervically infected animals, with titers ranging from 1:5 to 1:20. HIV-specific IgG antibodies could be detected in saliva from half of the systemically infected group, the highest titer being 1:2560, whereas the highest anti-HIV IgG titer in the mucosally infected group was 1:20. Western blot analyses of the first saliva samples obtained after initial virus exposure revealed IgG, IgA, and IgA subclass antibodies directed at the env, gag, or pol gene products in both groups of chimpanzees. Examination of IgG, IgA, IgA1, and IgA2 concentrations in vaginal washes from cervically infected animals showed that IgG levels were highest, but IgA and IgA subclass reactivities against HIV-1 were more prominent than that of IgG. These results demonstrate that systemic infection of chimpanzees with HIV-1 elicits mucosal responses specific for HIV, and vaginal infection of chimpanzees induces a common mucosal immune response reminiscent of that in humans. PMID- 9339845 TI - Mutations of the HIV type 1 V3 loop under selection pressure with neutralizing monoclonal antibody NM-01. AB - Variants of human immunodeficiency virus type 1 (HIV-1) were selected for resistance to the neutralizing monoclonal antibody (MAb) NM-01. MAb NM-01 recognizes the center of the third hypervariable domain (V3 loop) of the envelope gp120, and neutralizes diverse HIV-1 strains. In the continuous presence of MAb NM-01, transmission and propagation of molecularly cloned HIV-1 were performed in vitro to isolate escape variants. The polymerase chain reaction-single-strand conformation polymorphism and sequence analyses of these variants indicated that the antigenic change against MAb NM-01 is due to a single base substitution resulting in one amino acid interchange within the recognition site of MAb NM-01 in the V3 loop. Mutational analyses also demonstrated a nonrandom event of variability and the existence of mutational hot spots in the V3 loop. The bias of variability could be interpreted by the specificity of error-prone replication by HIV-1 reverse transcriptase. Furthermore, the results suggest that distribution of mutability might correlate closely with the stability of the secondary structure of RNA encoding the V3 loop region. PMID- 9339846 TI - The V1/V2 region of human immunodeficiency virus type 1 modulates the sensitivity to neutralization by soluble CD4 and cellular tropism. AB - A primary isolate (KMT) of human immunodeficiency virus type 1 (HIV-1) resistant to recombinant soluble CD4 (rsCD4) was isolated from an HIV-1-infected individual and grown in a T lymphoid cell line. KMT isolate passaged on CEM cells (KMT/CEM) was still resistant to rsCD4. The V1/V2 and V3 regions of the viral envelope glycoprotein are thought to be involved in various biological phenotypes. To determine the exact envelope region of the KMT isolate responsible for sensitivity to rsCD4 and cellular tropism, we performed sequence analysis of KMT and KMT/CEM isolates. Sequence analysis of the KMT isolate showed that the sequence of the V3 region was relatively homogeneous, whereas a considerable heterogeneity of the V1/V2 region was noted. In contrast, the sequences of the V1 to V3 regions were homogeneous in KMT/CEM isolates. Analysis of NL4-3-based recombinant viruses with amplified sequences of the V1 to V3 regions from KMT and KMT/CEM isolates showed that the V1/V2 region modulated the sensitivity to rsCD4. A change in resistance to rsCD4 by the V1/V2 region was associated with the ability of the isolate to replicate in macrophages and efficiently replicate in T lymphoid cell lines. A change to an isolate sensitive to rsCD4 was associated with reduced replication efficiency in T lymphoid cell lines. Our results suggest that the V1/V2 region is involved in modulating the sensitivity to rsCD4, macrophage tropism, and replication efficiency in T lymphoid cell lines. PMID- 9339847 TI - Longitudinal study of HIV-specific cytotoxic lymphocytes in HIV type 1-infected patients: relative balance between host immune response and the spread of HIV type 1 infection. AB - To evaluate the contribution of a specific cytotoxic response in the control of HIV infection in relation to clinical status, we performed serial analysis of anti-Env and anti-Gag cytotoxic activity in 13 infected individuals over a 6- to 10-year period, using cryopreserved peripheral blood mononuclear cells (PBMCs). Autologous EBV-transformed B cell lines infected in vitro with recombinant vaccinia viruses expressing HIV-1 env and gag genes were used as targets. Without any stimulation of the effector cells, we were able to show an anti-HIV cytotoxic activity in the PBMCs of 12 of 13 HIV-1-infected patients, consistent with chronic immune activation in HIV infection. Different patterns of HIV-specific cytotoxic activity were observed, and the extent of this cytotoxic response varied between the clinically defined groups of individuals. No direct relationship was observed with the number of CD4 and CD8 lymphocytes during the observation period. However, patients who remained asymptomatic had a more vigorous cytotoxic response than patients with clinical deterioration during the observation period, and a significant difference was observed for HIV Gag specific CTL activity. From these data, we suggest that the HIV-specific cytotoxic response has a protective role in the course of HIV infection. PMID- 9339848 TI - HIV type 1 envelope glycoprotein 120 carboxy-terminal peptide-induced human T cell lines selectively suppress heterogeneous proliferative T cell responses to soluble antigens. AB - It has been proposed that the highly conserved human immunodeficiency virus type 1 (HIV-1) envelope gp120 carboxy-terminal sequence, TKAKRRVVEREKR (CT120), may represent a functional mimic of the human leukocyte antigen (HLA) class II DR beta-chain third hypervariable region (HVR3) sequence motif located at position 69-81. Presentation of this potentially pathogenic fragment by HLA class I and/or II molecules, in a manner analogous to the indirect pathway of allorecognition, may induce both widespread cellular activation and also break self-tolerance, resulting in the selective and progressive anti-self HLA class II-directed immune suppression, which is a central feature of HIV-1 infection and the associated acquired immune deficiency syndrome (AIDS). To investigate the functional role of the HIV-1 gp120 C-terminal fragment T cell lines (TCLs) were raised from three healthy HIV-1-seronegative subjects at low risk of HIV-1 exposure, by repeated stimulation with a short synthetic 13-mer CT120 peptide in vitro. Graded concentrations (10[3] to 5 x 10[4]) of CT120 TCLs suppressed the primary 6-day proliferation of autologous PBMCs in response to the soluble antigens tetanus toxoid (TT) and purified protein derivative (PPD). In contrast, CT120 TCLs demonstrated no suppressive effect on 3-day phytohemagglutinin (PHA), concanavalin A (ConA), and pokeweed mitogen (PWM) mitogenic responses. Fractionation of CT120 TCLs into highly purified CD4+ and CD8+ T cell subsets demonstrated that the CD8+ T cell fraction mediated the suppressor effector function. HLA restriction analysis revealed a complex pattern as both anti-HLA class II DR and anti-HLA class I (A, B, C) MAbs inhibited proliferation of oligoclonal CD8+ CT120 TCLs. Strategies aimed at specifically inhibiting such putative immunopathogenic HIV-1-encoded T cell epitopes may be an important consideration for development of future HIV-1 immunotherapy. PMID- 9339850 TI - Inhibition of FIV replication by a ribozyme that targets the Rev response element. AB - The feline immunodeficiency virus (FIV) Rev protein and its cognate sequence the Rev response element (RRE) are essential for virus replication. Thus, the inhibition of either Rev or RRE function can significantly inhibit FIV replication. In the present study, we constructed a ribozyme that targets the RRE sequence and determined its ability to inhibit FIV replication. The RRE ribozyme cleaved the target molecule both in vitro and in FIV-infected cells. Furthermore, FIV replication was reduced significantly in the presence of the RRE ribozyme. FIV provides a good animal model system with which to develop novel antiviral strategies for the human immunodeficiency virus. PMID- 9339849 TI - Selenium supplementation suppresses tumor necrosis factor alpha-induced human immunodeficiency virus type 1 replication in vitro. AB - Selenium is a nutritionally essential trace element that is important for optimal function of the immune system. It is incorporated into selenoproteins as the amino acid selenocysteine and it is known to inhibit the expression of some viruses. In this study, we show that selenium supplementation for 3 days prior to exposure to tumor necrosis factor alpha (TNF-alpha) partially suppresses the induction of human immunodeficiency virus type 1 (HIV-1) replication in both chronically infected T lymphocytic and monocytic cell lines. In acute HIV-1 infection of T lymphocytes and monocytes in the absence of exogenous TNF-alpha, the suppressive effect of selenium supplementation was not observed. However, selenium supplementation did suppress the enhancing effect of TNF-alpha on HIV-1 replication in vitro in acutely infected human monocytes, but not in T lymphocytes. Selenium supplementation also increased the activities of the selenoproteins, glutathione peroxidase (GPx) and thioredoxin reductase (TR), which serve as cellular antioxidants. Taken together, these results suggest that selenium supplementation may prove beneficial as an adjuvant therapy for AIDS through reinforcement of endogenous antioxidative systems. PMID- 9339851 TI - Fibronectin modulates endothelial response to HIV type 1 Tat. AB - The normal function of the endothelium is impaired in HIV-1 infection. Disturbances of the local cytokines as well as the release of HIV-1 Tat by infected mononuclear cells play a role in endothelial dysfunction. We studied the effects of Tat on the human endothelial ECV cell line. In this system, Tat inhibited cell proliferation only in the presence of fibronectin as a culture substrate, whereas it did not modulate plasminogen activator activity, cell migration, or synthesis of fibronectin. Because amino acids 49-57 contains a nuclear translocation sequence, we also evaluated the potential intracellular role of Tat in tat-transfected ECV cells. tat transfectants showed inhibition of cell growth, unaffected cell migration and plasminogen activator activity, and a significant induction of the expression of fibronectin. PMID- 9339853 TI - gag and env sequences of an A/G/H recombinant from a Zairian HIV type 1 isolate. PMID- 9339852 TI - Genetic variability of HIV type 1 in Honduras. PMID- 9339854 TI - The integration of medical management with recovery. AB - Patients recovering from alcohol and other drug addiction have unique medical and pharmacological needs. Careful selection of medications call decrease the risk of relapse. Angiotensin-converting enzyme inhibitors and calcium channel-blocking medications are excellent choices to treat hypertension. Most gastrointestinal problems resolve with abstinence and can be treated nonpharmacologically. In managing pain, physicians should avoid narcotics and use nonpharmacological treatment whenever possible. Treating recovering patients with HIV can be challenging because of the side effects of many of the antiviral medications. The newer antiviral agents have fewer side effects and contraindications. Commonly used remedies for colds and cough can cause a relapse to drug use. Patients with diabetes mellitus need to be monitored very closely in early recovery to prevent hypoglycemia. Frequently a team approach is helpful in managing the medication needs of patients in recovery. PMID- 9339855 TI - Integration of treatment and posttreatment variables in predicting results of abstinence-based outpatient treatment after one year. AB - A multi-site, longitudinal study of patients undergoing outpatient alcohol and drug dependence treatment was conducted in private outpatient facilities, consisting of 2,029 subjects from 33 independent programs enrolled in a national addiction treatment outcomes registry. Pretreatment demographic and substance variables, treatment utilization variables, and post-treatment continuum of care variables were examined simultaneously in a multivariate prediction context for association with outcome. Upon admission patients provided history information to treatment staff trained in the collection of data for the evaluation efforts. Trained interviewers conducted consecutive structured interviews prospectively for treatment outcome at six- and 12-month follow-up periods. Multivariate analysis with stepwise multiple regression indicated that, relatively speaking, the most powerful predictors of treatment outcome were posttreatment variables: namely, support group attendance and involvement in a continuing care program. Pretreatment and treatment variables contributed proportionately little to the prediction of outcome. Additional sequential-stage analysis showed that the incremental contribution to prediction by posttreatment attendance at Alcoholics Anonymous and involvement in a treatment program following discharge far exceeded the initial predictive validity of the 14 pretreatment and treatment variables examined. Participation in posttreatment continuing care correlated with statistically significant reductions in job absenteeism, inpatient hospitalizations, and arrest rates. Posttreatment more than pretreatment factors may be decisive in influencing risk for relapse. PMID- 9339856 TI - The integration of pharmacological therapy for comorbid psychiatric and addictive disorders. AB - The integration of pharmacological therapies for comorbid disorders requires an acceptance of independence and interactions of respective addictive and psychiatric disorders. At the same time, alcohol and other drugs induce psychiatric states that are indistinguishable from psychiatric disorders. On the other hand, while psychiatric disorders do not induce addictive use of alcohol and drugs, they do pose vulnerabilities to the development of addictive disorders. Generally, the treatment of comorbid disorders begins with abstinence and evaluation of the effects of alcohol and other drugs in contributing to the psychiatric picture. In the case of comorbid disorders, stabilization and standard treatments can be employed with certain cautions, namely, to avoid the use of addicting medications such as benzodiazepines and opiates beyond the detoxification stage. High potency neuroleptics and antidepressants, particularly selective serotonin reuptake inhibitors (SSRIs) can be used to treat continuing psychiatric states after the exclusionary criteria in DSM-IV for substance related disorders have been applied to the clinical case. If the psychiatric symptoms clear with sustained abstinence, little or no medications may be required. Specific treatment of the addictive disorders will often determine the extent that addictive disorders are responsible for psychiatric symptomatology. Alternatively, treatment of the psychiatric disorder will enhance compliance with addiction treatment. PMID- 9339857 TI - Integration of health care economics for addiction treatment in clinic care. AB - Over the past decade, radical changes have occurred in the availability and funding of treatment services for individuals with addictive disorders. Traditional inpatient and residential treatments have been replaced by outpatient settings. "Standard" addiction treatments, such as 28-day inpatient hospitalizations, are largely a thing of the past. The process of treatment planning has also changed. Treatment decisions now depend on third-party coverage rather than severity of illness. The procedures for determining eligibility and obtaining authorization for treatment are often ponderous and impersonal and determined through administrative procedures that are external to the treatment facility. The criteria used to determine the type of treatment seem arbitrary rather than clinically based. PMID- 9339858 TI - Integration in education for addiction medicine. AB - Addiction to alcohol and other drugs is a serious public health problem that is one of the most common disorders seen in medical practice. Although it is an extremely common disorder, it is poorly diagnosed and treated by physicians. In order to begin to develop an integrated approach to education and addiction, one must define the many roles of the physician working with addicted patients. Training about addictions must begin early in the medical student's career, and continue in a vertically integrated way throughout medical school. The notion of addiction as a disease process must be introduced and integrated into course materials in the preclinical years. Careful attention must be paid to the development of positive views toward working with addicted patients, and students must be indoctrinated early with the idea that physicians have a responsibility to diagnose and manage addicted patients. Students should be given multiple opportunities to learn and use screening interviews for addiction in preclinical interviewing courses, and while on the clerkships. Residency education and continuing medical education in addictions are also important, so that faculty may become good role models for students in this critical area. PMID- 9339859 TI - Addiction medicine: a place for faculty development. AB - Addiction medicine must fight for its space in the undergraduate, graduate and continuing medical education curricula, as do most other clinical domains. Often curriculum time is provided to specialty areas when a clear relevance to the overall curriculum is made obvious. Increasing the awareness of addiction medicine through institutionalized faculty development programs can serve to foster the integration of this specialized curriculum. Institutionalizing faculty development is proposed in a description of a four-phase model. Specific recommendations of goals, processes, and critical steps in the faculty development process supporting scholarship leading to curricular change are described. PMID- 9339860 TI - Addiction medicine and continuing medical education. AB - Addictive disorders are one of the most common problems encountered by primary care physicians. In the last decades there has been a significant effort by organizations, universities, and private foundations to increase the teaching of alcohol and drug abuse issues to medical students, residents and practitioners. Still, up to now, the subject has not been presented appropriately at either the undergraduate or graduate medical education level and the majority of physicians in practice have not been adequately instructed in addiction medicine. This article reviews the literature on addictive disorders and medical education, exploring issues concerning continuing medical education (CME) in particular. The authors discuss the problems relative to this subject and the educational techniques and methods most appropriate to changing attitudes and behaviors of physicians. They also design an approach to a CME program on addictive disorders for primary care physicians that incorporates multiple teaching/learning methodologies. PMID- 9339861 TI - Temporal patterns of veterans' psychiatric service utilization, disability payments, and cocaine use. AB - This study examined temporal patterns of service utilization, disability benefits, and substance use. Specifically, it investigated whether the first day of the first week of each month (when disability payments are disbursed) was associated with increased emergency room (ER) use and more frequent cocaine use among psychiatric patients. All 1993 psychiatric ER presentations (n=1,448) at a Veterans Administration hospital were reviewed in order by the week of each month in which they occurred. A random subsample of only those admitted to an inpatient psychiatric service (n=143) was further assessed for amount of disability payments received and recent cocaine use. This study found that for the total population of patients utilizing the ER, most ER visits occurred during the first week, followed by weeks two, three, and four respectively. The highest percentage (49%) of patients who used cocaine were those admitted during the first week of the month, followed by week two (39%), week four (28%) and week three (25%). For the subsample of patients admitted to inpatient services, patients hospitalized during the fourth week of the month were those receiving the highest disability payments. This study found that cocaine users have the most ER visits during the first week of the month following receipt of benefits. Current data, if confirmed, would suggest public policy changes, such as payment of entitlement money to cocaine users through a third-party payee and stipulated treatment for psychiatric patients with substance use disorders as a condition of payment. Ethical and political issues, including confidentiality and patient autonomy, would need to be considered in any such policy changes. PMID- 9339862 TI - Substance abuse treatment for "hazardous users": an early intervention. AB - A six-session cognitive behavioral protocol has been developed for substance abusers who meet the description "hazardous users." This category includes individuals evidencing mild to moderate use of alcohol or other drugs, whose lifestyles are minimally disrupted, or who are displaying signs of problem use or abuse, but are unwilling to enter intensive treatment. The treatment model is nonconfrontational and is designed to motivate the individual to recognize the problems associated with his or her substance use and initiate treatment-seeking behavior. The intervention may be particularly useful in situations where employees have tested positive for substances but deny having a problem, where friends or family members report help is needed but the individual denies any problem, or where an alcohol or other drug problem is clearly evidenced but the individual doesn't acknowledge a problem. A positive outcome is indicated by the client taking action which is consistent with an increased awareness of the problem as conceptualized by Prochaska and DiClemente (1982). This model is an alternative to the traditional confrontational models of "breaking through denial." The philosophies employed by William Miller and associates and by the Matrix treatment models form the basis of the intervention. PMID- 9339863 TI - "Enhancing the quality of our endeavors": the Farmington Consensus. PMID- 9339864 TI - Prevalence of childhood epilepsy in Kaunas, Lithuania. AB - The prevalence of epilepsy in children aged 0-15 years of Kaunas city, Lithuania, was evaluated on 1 January 1995. Multiple sources for case identification were used, i.e. medical records at the university hospital, regional outpatient clinics and consultation centres, institutions, schools and kindergartens for the handicapped. Active epilepsy was defined as two or more unprovoked epileptic seizures with at least one seizure occurring within the previous 5 years, regardless of the antiepileptic drug treatment. Prevalence was found to be 4.25 (3.42, if age-standardized) in 1000. The highest rate was found in the 10-14 years age group. The male/female ratio was 1.29. No possible causes could be determined in 60.3% of cases. Congenital causes were diagnosed in 18.8% of cases, perinatal causes in 15.3%, traumatic causes in 2.6% and neuroinfectious causes in 2.4%. Classification of epilepsies and epileptic syndromes [Commission on Classification and Prognosis of the International League Against Epilepsy. Proposal for revised classification of epilepsies and epileptic syndromes. Epilepsia 1989; 30:389-399] revealed that 50% of cases were localization-related epilepsies, 29.9% were generalized epilepsies, 15.9% were undetermined whether partial or generalized and 4.2% were unclassifiable. Rates for idiopathic, symptomatic and cryptogenic cases are presented. PMID- 9339865 TI - A developmental expression of AMPA-selective glutamate receptor subunits in human basal ganglia. AB - The development of AMPA-selective glutamate receptors (GluR1 and GluR2-3) in human basal ganglia (BG) was investigated in 23 normal brains by means of an immunohistochemical method. Immunoreactivity to GluR1 and 2-3 was detected in the cytoplasm and dendrites of small and large neurons in the BG. GluR2-3 immunoreactivity-positive neurons were clearly observed at 23-24 gestational weeks (GW) in the globus pallidus and at 32 GW in the neostriatum, and reached a peak at 32 GW and 39 GW, respectively. GluR2-3 positive neurons in the BG began to decrease at 1-4 months of age, reaching the low level of adults by 7 months of age. The developmental pattern of GluR1 was similar to that of GluR2-3 in the BG, but the immunoreactivity to GluR1 was a little weaker than that of GluR2-3 in the neostriatum. Furthermore, GluR1 and 2-3 subunit Western blotting confirmed the specificity of the immunohistochemistry. Our results suggest that the development of GluR1 and 2-3 in the BG is consistent with neuronal development in the BG, which supports further that GluR1 and 2-3 play an important role in neuronal differentiation and maturation of the BG. PMID- 9339866 TI - The influence of feeding patterns on head circumference among Turkish infants during the first 6 months of life. AB - The influence of various feeding patterns on physical growth and mental development of the infant, particularly during the first 6 months of life, is an important subject. Head circumference values of 172 healthy new-born infants were included in the study; 62 were exclusively breast-fed (BF), 58 were mixed-fed (MF) and 52 were formula-fed (FF). No significant differences were found in head circumference values between the groups at birth (BF 35.2+/-0.1, MF 35.1+/-0.1, FF 35.0+/-0.1 cm for boys and BF 35.0+/-0.1, MF 34.9+/-0.1, FF 34.8+/-0.1 cm for girls). At the end of the first month, the infants in the BF group (38.3+/-0.1 cm and 37.9+/-0.1 cm for boys and girls, respectively) had strikingly greater head circumference measurements than the others (MF 36.7+/-0.1, FF 36.6+/-0.1 cm for boys and MF 36.5+/-0.1, FF 36.4+/-0.1 cm for girls) (P < 0.05). However, in the subsequent 4-month period, the values detected in each group were almost the same. At the sixth month, head circumference-for-age values of infants in MF (42.6+/-0.1 cm for boys, 41.4+/-0.1 cm for girls) and FF (42.5+/-0.1 cm for boys, 41.5+/-0.1 cm for girls) were well below those of BF group (43.7+/-0.1 cm and 42.9+/-0.1 cm for boys and girls, respectively) and the standard curve (P < 0.05). These results suggest that exclusive breast feeding is sufficient during the first 6 months, the most important period of life. PMID- 9339867 TI - Lamotrigine as add-on drug in children and adolescents with refractory epilepsy and mental delay: an open trial. AB - We report the results of an open trial with lamotrigine (LTG) as add-on drug in children and adolescents with refractory epilepsy and mental delay. Thirty-seven outpatients received LTG for a median period of 7 months at a daily dose of 5 and 15 mg/kg in valproate and non-valproate patients, respectively. The total number of seizures decreased by 100% in eight patients (21.6%) and by >50% in five patients (13.5%). However, the number of seizures remained unchanged in 20 patients (54.1%) and increased in four (10.8%). Lamotrigine was more effective in patients with typical and atypical absences, and in patients affected by atonic seizures. Six children (16.2%) developed generally mild adverse side-effects suggesting that LTG is well tolerated. PMID- 9339868 TI - Therapeutic doses of theophylline exert proconvulsant effects in developing mice. AB - We studied the effect of therapeutic doses of theophylline on electrically induced convulsions in developing mice. A theophylline dose as small as 3 mg/kg increased seizure susceptibility of 21-day-old mice, but not of 42-day-old mice. These findings were consistent with clinical reports that theophylline at the therapeutic blood concentrations occasionally induced convulsions in children. The age-dependent proconvulsant effect of theophylline was well inhibited by phenobarbital (PB), dose-dependently, but not by other well-established antiepileptic drugs (AEDs). PB may be a good choice of AED in patients with bronchial asthma and seizure disorders, if PB is indicative for their seizure types. The proconvulsant effect of theophylline in 21-day-old mice was counteracted by not only an adenosine A1 agonist, but also an NMDA antagonist and a histamine H3 antagonist. Several studies have established that the proconvulsant effect of theophylline intoxication is mainly due to the blockade of adenosine A1 receptors. The present findings suggested that the proconvulsant properties of therapeutic doses of theophylline in developing period were different from those of theophylline intoxication. Combination of therapeutic doses of theophylline and centrally-acting histamine H1 antagonists showed proconvulsant effects even in 42-day-old mice, suggesting that peripherally acting histamine H1 antagonists, such as astemizole, evastine and epinastine, were much safer than centrally acting histamine H1 antagonists for patients with both allergy and seizure history. PMID- 9339869 TI - Childhood occipital epilepsy: seizure manifestations and electroencephalographic features. AB - Childhood epilepsy with occipital paroxysms (CEOP) is an idiopathic localization related epilepsy. A typical seizure in CEOP begins with visual symptoms, followed by hemiclonic seizures, complex partial seizures or generalized tonic-clonic seizures. Benign nocturnal childhood occipital epilepsy (BNCOE), characterized by nocturnal seizures with tonic deviation of the eyes followed by vomiting, has the same electroencephalographic features as CEOP. In this study, we report the seizure symptoms and electroencephalographic features of 21 cases with CEOP or BNCOE. Out of these patients, nine had BNCOE, six had CEOP, four had CEOP and BNCOE and the remaining two belonged to the incomplete syndrome because of no paroxysmal discharges in EEG. When the patients with BNCOE awoke from sleep, they had tonic deviation of the eyes and could describe visual symptoms. Patients with CEOP had seizures beginning with visual symptoms followed by loss of consciousness but no generalized convulsions. In three cases, in addition to the occipital spikes, independent centro-temporal spikes were recorded and in another three cases generalized spike-wave discharges were recorded. Such a combination suggests the idiopathic nature of these epilepsies. We concluded that in the diagnosis of CEOP and BNCOE, the seizure symptomatology is important even if the EEG can be considered normal. PMID- 9339870 TI - Intracranial calcifications, epilepsy, and optic atrophy associated with metaphyseal dysplasia: a case report. AB - A 15-year-old boy presenting with epilepsy, optic atrophy and intracranial calcifications was diagnosed as having metaphyseal dysplasia by bone X-ray examinations. The patient had no laboratory data suggesting other metabolic or endocrinologic disorders. In addition, CT scans showed unique intracranial calcifications of the corpus callosum and periventricular and subcortical white matter, which were distinct from those of previously reported disorders. This case may represent a unique subset or a new type of metaphyseal dysplasia associated with intracranial calcifications and central nervous system symptoms. PMID- 9339871 TI - Gelastic epilepsy: video-EEG, MRI and SPECT characteristics. AB - Gelastic epilepsy, or ictal laughter, is a relatively uncommon type of seizure which may occur singly or, more frequently, with other types of convulsions. Gelastic seizures have been observed to be associated with many different conditions, mainly hypothalamic hamartomas. We report on a patient whose ictal laughter was the only neurologic disturbance. Ictal video-EEG demonstrated seizure arising from the left frontal region with subsequent involvement of the contralateral homologous area and secondary generalization. MRI showed an enlarged left frontal horn of the lateral ventricle. Postictal SPECT, performed 6 min after the seizure had ended, showed hypoperfusion in the bilateral frontoparietal region and in both cerebellar hemispheres; the presence of this abnormality may be due to the spreading of the cortical epileptogenic focus and to the complex intercommunication between the frontal cortex and the cerebellar hemispheres. Interictal SPECT, in accordance with MRI features, demonstrated a left frontoparietal hypoperfusion. The neurofunctional features observed in the reported child could suggest that gelastic epilepsy originates in the frontal cortex. However, further studies are undoubtedly needed to define the pathogenetic mechanisms of ictal laughter. PMID- 9339872 TI - Spontaneous remission of childhood epilepsy in two patients with focal extraopercular cortical dysplasia. AB - Childhood-onset partial epilepsy caused by focal cortical dysplastic lesions (FCDLs) is often severe. A few patients reported with a favorable outcome had a normal neuropsychological examination, and FCDLs were always localized around the opercular region, suggesting that extent and location of the lesion may account for the favorable outcome. We report two patients with extraopercular FCDLs, who had a spontaneous remission of their childhood-onset epilepsy, despite a severe neurological deficit. A 22-year-old girl (patient 1) and a 16-year-old boy (patient 2), began to have partial seizures at the age of 9 years and 1 year respectively. On neurological examination, patient 1 had left hemiparesis and patient 2 had low IQ. Interictal EEG recordings revealed repetitive epileptiform discharges involving the right temporo-parietal or frontal areas in patients 1 and 2 respectively. MRI study showed focal cortical thickening or abnormal gyration located over the right parietal and frontal region respectively in patients 1 and 2, but failed to evidence T2 prolongation in the white matter beneath the dysplastic cortex. Optimal antiepileptic regimen always stopped seizures. Their long-term course was favorable, with remission of the seizures and normalization of EEG recordings, even 4-5 years after medication withdrawal. In conclusion, FCDLs may cause epilepsy with a benign course even in patient with mental retardation or neurological abnormalities. This may be related to a morphologically milder dysplastic lesion than found in patients with FCDLs and severe epilepsy. PMID- 9339873 TI - Telencephalosynapsis (synencephaly) and rhombencephalosynapsis with posterior fossa ventriculocele ('Dandy-Walker cyst'): an unusual aberrant syngenetic complex. AB - Agenesis of the cerebellar vermis (paleocerebellar agenesis) with fusion of the cerebellar hemispheres (rhombencephalosynapsis) is a rare malformation of the central nervous system (CNS). Its combination with synencephaly (telencephalosynapsis), telencephalic ventricular aplasia, aqueductal atresia and cystic fourth ventricle has not yet been described, as far as we know. Here, we report this combination in a 23-weeks' gestation male fetus who was aborted to a 24-year-old diabetic mother. In this fetus with cerebral and cerebellar hemispheric fusion, vermian agenesis was associated with a Dandy-Walker-like posterior fossa cyst, in spite of the fusion of the hypoplastic cerebellar hemispheres. The CNS malformations were further accompanied by dysmorphic facial stigmata such as unilateral atresia of the external ear, ocular hypertelorism and a broad nasal bridge. Preaxial polydactyly and contractures of the upper limbs were the only associated non-cranial abnormalities. Cytogenetic studies revealed a numerically and structurally normal male (46, XY). The malformation complex described in this fetus of a mother with antedating pregnancy diabetes appears to represent a previously undescribed aberrant syngenetic CNS phenotype, some basic teratogenetic aspects of which will be discussed in this paper. PMID- 9339874 TI - Two cases of essential myoclonus, epilepsy, mental retardation and anxiety disorders. AB - We examined an 18-year-old female and an 18-year-old male with mild mental retardation who suffered from the oscillatory form of sporadic essential myoclonus from an age of 3 years. Although the generalized oscillatory myoclonus resembled severe essential tremor, surface electromyography revealed small myoclonic jerks with frequencies of 6-8 Hz. As concomitant symptoms, the female case exhibited overanxious irritability from early childhood and generalized epileptic seizures occurred from the age of 4 years. In the male case, an obsessive-compulsive disorder and photosensitive convulsive seizures were persistently noted from early childhood. All their symptoms had been stable for at least the last 10 years. Thus, although non-progressive tremulous movements are rare in early childhood, sporadic essential myoclonus is causative. In contrast to hereditary essential myoclonus, sporadic essential myoclonus is considered to be more heterogeneous, especially in the various associated symptoms. PMID- 9339875 TI - Pontine hypoplasia in a child with sensorineural deafness. AB - A 2-year-old girl with bilateral sensorineural deafness showed pontine hypoplasia as well as a bulging contour of the pontine tegmentum on magnetic resonance imaging (MRI). There were no bilateral responses of brainstem auditory-evoked potentials (BAEPs). The absent late components of blink reflex (BR) indicated brainstem dysfunction. Chromosomal abnormalities and neurodegenerative or neurometabolic disorders, which might have been the cause of the pontine hypoplasia, were ruled out. The authors describe a rare case with pontine hypoplasia combined with sensorineural deafness and absent blink reflex and suggest that the brainstem in this child may become involved in the early gestation period. PMID- 9339876 TI - Prospective study of ankle clonus at the first year of life. PMID- 9339877 TI - Lifetime diagnosis of major depression as a multivariate predictor of treatment outcome for inpatients with substance use disorders from abstinence-based programs. AB - A multisite, longitudinal study of patients undergoing inpatient alcohol and drug dependence treatment was conducted in private inpatient facilities, consisting of 4339 subjects from 38 independent programs enrolled in a national addiction treatment outcomes registry. Structured interviews were conducted upon admission, including documentation of current alcohol/drug disorder (DSM-III-R) and lifetime diagnosis of major depressive syndrome; structured interviews were conducted prospectively at 6- and 12-month follow-up periods. The prevalence rate of lifetime diagnosis of major depression in the sample was 39%. Comorbidity varied according to gender and substance of choice. Lifetime depressive symptoms did not correlate with differential length-of-stay, treatment completion, or follow-up consent and, at best, were very weakly associated with follow-up contact. Patients diagnosed with lifetime depression showed the same frequency of participation in posttreatment continuing care: they also showed statistically significant reductions in job absenteeism, inpatient hospitalizations, and arrest rates pre- vs. posttreatment comparable to those of patients without lifetime depression diagnosis. Lifetime major depressive syndrome was not a predictor of outcome in response to abstinence-based treatment. Involvement in posttreatment continuing care accounted for far greater outcome variance. Posttreatment vs. pretreatment factors may be more decisive in influencing risk for relapse. PMID- 9339878 TI - Impact of shorter lengths of stay on status at discharge in bipolar mania. AB - This study assesses the impact of shortening the inpatient length of stay on status at discharge in patients with mania. METHODS: The authors performed a chart review on 131 patients with discharge diagnoses of bipolar disorder, current episode manic type, admitted to the private attending service at Colorado Psychiatric Hospital between 1985 and 1995. In 1990, a new program (the alternatives program) that provides a continuum of acute care services and shorter inpatient lengths of stay was instituted. Retrospectively assessed GAF, CGI, treatment outcome rating scores, and length of stay (LOS) were compared for the prealternatives (1985-1989), early alternatives (1990-1992), and recent alternatives (1993-1995) program treatment eras. A progressive decrease in inpatient LOS, duration of the acute care episodes, and total service utilization was seen across eras. Despite the more recent shortening in LOS, no significant differences were seen in GAF and treatment outcome rating scores at discharge. GAF and treatment outcome rating scores on hospital days 3 and 7, however, suggested that patients were improving more rapidly in the more recent eras. Inpatient LOS and duration of the acute care episodes have significantly decreased over the last 10 years, but patients appear no more ill at discharge. The authors postulate that changes in psychopharmacologic practice and the inpatient treatment model may have facilitated the more rapid clinical improvement seen in the more recent eras. The authors caution that we need prospective studies that include postdischarge follow-up to assess further the impact of shorter inpatient stays on the posthospital course of manic patients. PMID- 9339879 TI - Divalproex: a possible treatment alternative for demented, elderly aggressive patients. AB - Elderly, demented people often exhibit behavioral dyscontrol. Divalproex appears to be safe and effective in the management of this presentation. Twelve cases treated with divalproex all responded with improved emotional control. Patients became less verbally and physically disruptive and much more socially appropriate. Divalproex was well tolerated in this population with none of the subjects experiencing significant medicinal side effects. This uncontrolled report suggests that divalproex should be considered as a pharmacotherapy for aggressivity in cognitively impaired, elderly people. PMID- 9339880 TI - Explained and unexplained medical symptoms in generalized anxiety and panic disorder: relationship to the somatoform disorders. AB - We have examined the numbers and types of symptoms in a sample of 90 patients with generalized anxiety disorder (GAD) and 77 patients with panic disorder (PD) collected from six different sites during the conduct of a multicenter clinical trial. This information was obtained utilizing the Health Questionnaire, a 47 item self-report list of medical symptoms, patterned after the Somatization Disorder section of the Diagnostic Interview Schedule. Although the patients in this sample had a wide variety of medically explained and unexplained physical symptoms, none of them qualified for a diagnosis of somatization disorder by DSM III-R criteria. GAD and PD patients reported remarkably similar numbers of explained and unexplained medical symptoms. The panoply of somatic symptoms presented by these patients presents a formidable diagnostic challenge for clinicians. These findings suggest that the pattern of overutilization of medical services that is well documented for PD patients may also be found for GAD patients. PMID- 9339881 TI - Once-daily venlafaxine extended release (XR) and venlafaxine immediate release (IR) in outpatients with major depression. Venlafaxine XR 208 Study Group. AB - This was a randomized, double-blind, placebo-controlled comparison of the efficacy and safety of once-daily venlafaxine extended release (XR) and venlafaxine immediate release (IR). Outpatients with DSM-III-R major depression were randomly assigned to venlafaxine XR, 75 mg once daily, venlafaxine IR, 37.5 mg twice daily, or placebo for a maximum of 12 weeks. If the response was inadequate after 2 weeks of treatment, the dosage of venlafaxine XR or IR could be increased to 150 mg daily. The primary efficacy variables were the 21-item Hamilton Depression (HAM-D) Rating Scale total score and depressed mood item, the Montgomery-Asberg Rating Scale (MADRS) total scores, and the Clinical Global Impressions (CGI) severity scale. Two hundred seventy-eight patients were evaluated for efficacy. Venlafaxine XR was significantly superior (p < 0.05) to placebo beginning at week 2 for the HAM-D, week 3 for the MADRS, and week 4 for the CGI severity. Similarly, venlafaxine IR was significantly superior (p < 0.05) to placebo beginning at week 2 on the HAM-D total and depressed mood item, week 3 on the MADRS total, and week 6 on the CGI severity scales. Venlafaxine XR exhibited superiority (p < 0.05) over venlafaxine IR at week 12 for all efficacy variables. The most common treatment-emergent adverse event with venlafaxine XR was nausea. The incidence of nausea was highest during the first 2 weeks with a low likelihood of developing nausea thereafter. The results of this study indicate that venlafaxine XR is safe, effective, and well tolerated for the treatment of major depression at once-daily doses ranging from 75 to 150 mg. PMID- 9339882 TI - Clozapine withdrawal catatonia and neuroleptic malignant syndrome: a case report. AB - Catatonia as a clozapine withdrawal syndrome has not been documented. We report a case of excited catatonia with fever, autonomic instability, and delirium--a picture of malignant catatonia (lethal catatonia) after abrupt clozapine withdrawal. The use of conventional neuroleptics transformed the excited malignant catatonia into a stuporous state resembling neuroleptic malignant syndrome (NMS). Such a transformation of excited lethal catatonia into NMS has been described in the literature, providing support for the hypothesis that NMS is a variant of catatonia. Opinions, however, have been conflicting whether lethal catatonia and NMS are indistinguishable. We argue that NMS may be regarded as a neuroleptic-induced retarded (stuporous) subtype of malignant catatonia, clinically indistinguishable from nonneuroleptic retarded malignant catatonia but different from the excited form. To differentiate between the two subtypes of malignant catatonia would help resolve the controversy. The nosological status of excited catatonia, a poorly studied condition, remains unclear. The two subtypes of catatonia may differ in pathophysiology and responses to treatment. Clinicians should be alert to catatonia as a possible clozapine withdrawal phenomenon, and excited catatonia deserves more research attention. PMID- 9339883 TI - Response of obsessive compulsive disorder to carbamazepine in two patients with comorbid epilepsy. AB - An association between epilepsy and obsessive compulsive disorder (OCD) has been noted. The response of two patients with OCD and comorbid epilepsy to carbamazepine is reported. It is hypothesized that obsessive compulsive symptoms may be a variant of epileptiform forced thinking in a subgroup of patients, and may be preferentially responsive to anticonvulsant therapy. PMID- 9339884 TI - Psychosis during pregnancy: treatment considerations. AB - The onset of psychosis during pregnancy presents several difficult management decisions and a careful risk-benefit analysis is required. Withholding antipsychotic treatment may produce more risks than benefits. Studies on neuroleptic teratogenicity are contradictory. Most of the commonly used neuroleptics exhibit a pregnancy risk of category C. Neuroleptic use during pregnancy may be associated with adverse effects in the pre- and postnatal period. These concerns include compromising uterine blood flow, post-partum neonatal sedation, and extrapyramidal signs expressed in the neonate. Each neuroleptic exhibits a unique pharmacokinetic profile. The antipsychotic properties and side effects considered most significant include sedation, half life, hypotension, and apparent hydrophilicity. In this case study a decision to select molindone was based on these parameters. PMID- 9339885 TI - Risperidone and hyponatremia: a case report. AB - A 48 year-old white male not suffering from endocrine disease or polydipsia, not taking diuretics, and suffering from no renal disease was started on risperidone and discharged on no other drug from Western Missiouri Mental Health Center (WMMHC) after an 8-day hospitalization. Seven days later he was admitted to a university medical center with generalized seizures, hyponatremia, respiratory failure, and rhabdomyalysis. He eventually recovered, was transferred back to WMMHC, and stabilized on appropriate medication. A search of the literature indicates no case reports linking risperidone to hyponatremia. It is assumed that the mechanism of hyponatremia is similar to other psychotropic medication in that it is secondary to the syndrome of inappropriate antidiuretic hormone (SIADH). PMID- 9339886 TI - Chemical dependence involving glucocorticoids. AB - The abuse of anabolic steroids has been well documented in the literature. In contrast, relatively little is written about the abuse potential of glucocorticoids. A case report of a 51-year-old man with substance dependence on prednisone is reported. The limited available literature on the abuse of glucocorticoids is briefly reviewed, and the challenge of treating patients who abuse these medications discussed. PMID- 9339887 TI - Nucleolar snoRNA and ribosome production. PMID- 9339888 TI - Mutagenesis of the positively charged conserved residues in the 5' exonuclease domain of Taq DNA polymerase. AB - Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction method. Taq DNA polymerase has a domain at the amino terminus (residue 1 to 290) that has a 5' exonuclease activity and a domain at the C-terminus that catalyzes polymerase reaction. Taq DNA polymerase is classified into the pol I family which is represented by E. coli DNA polymerase I. The alignment of amino acid sequences for the 5' exonuclease domains of the pol I family DNA polymerases shows six highly conserved sequences called motifs A to F. Motif C contains three positively charged residues such as 74Arg, 82Lys and 85Arg which might be involved in catalysis. In order to understand the function of those residues, they are mutagenized to alanine. The 5' exonucleolytic activities of those mutated 5' exonucleases decreased by 80 to 90%, thereby implying that three positively charged residues play certain roles in the 5' exonuclease catalysis. PMID- 9339889 TI - H+-pumping ATPase has little stimulatory effect on in vitro translocation of a model protein into Vibrio alginolyticus inside-out membrane vesicles. AB - We studied the effect of a H+ electrochemical potential generated by F1F0 ATPase on in vitro translocation of a model protein into Vibrio alginolyticus inside-out membrane vesicles. The F1F0 ATPase of V. alginolyticus catalyzed the pumping of H+ coupled to ATP hydrolysis as measured in fluorescence quenching experiments. Consequently, this enzyme leads to the generation of a H+ electrochemical potential. The H+ electrochemical potential generated by F1F0 ATPase was completely abolished by 30 microM N,N'-dicyclohexylcarbodiimide (DCCD) or 5 microM carbonylcyanide m-chlorophenylhydrazone (CCCP) at 30 degrees C. The treatment of membrane vesicles with 30 microM DCCD at 30 degrees C had little influence on the translocation activity of uncleavable OmpF-Lpp, a model secretory protein, as compared to the intact membrane vesicles. On the other hand, the NADH:quinone oxidoreductase of V. alginolyticus is known to be a Na+ pump that leads to generation of a Na+ electrochemical potential. This Na+ electrochemical potential stimulates protein translocation into inside-out membrane vesicles prepared from V. alginolyticus in the presence of Escherichia coli SecA [Tokuda, H., Kim, Y. J., and Mizushima, S. (1990) FEBS Lett. 264, 10 12] From these results, it is evident that the stimulatory effect of the Na+ electrochemical potential on protein translocation in V. alginolyticus is not affected by the H+ electrochemical potential influence of F1F0 ATPase. PMID- 9339890 TI - Km genotyping using polymerase chain reaction-allele specific oligonucleotide in Koreans. AB - The allotype of human immunoglobulin kappa light chain constant region gene (IGKC) has been identified by the serological assay or DNA typing method. The three allotypes Km*1, Km*1,2 and Km*3 have amino acid substitutions at positions of codons 153 and 191 in the kappa light chain constant region. The Km allotypes were designated as follows: Km*1 has valine (GTC) at position 153 and leucine (CTC) at 191; Km*1,2 has Ala153 (GCC) and Leu191 (CTC); Km*3 has Ala153 (GCC) and Val191 (GTC). In this study, polymerase chain reaction-allele specific oligonucleotide (PCR-ASO) was used to provide a specific allelic typing system for the IGKC gene in 213 unrelated Koreans. For Km genotyping, the IGKC coding regions plus 35 bp of the 3' untranslated regions were amplified with specific primers. Amplified genes were hybridized with Km ASO probes at positions of codons 153 and 191 and were detected using a colorimetric detection method. The frequencies of Km*1,2/Km*1,2, Km*1,2/Km*3, and Km*3/Km*3 were 17.8% (n=38), 51.7% (n=110), and 30.5% (n=65), respectively. The estimated allele frequencies for Km*1,2 and Km*3 were 0.44 and 0.56, respectively. The homozygote or the heterozygote of Km*1 was not observed in this study. The PCR-ASO method distinguished the alleles Km*3, Km*1,2, and Km*1 and also distinguished the homozygote from heterozygote. Km frequencies vary significantly among various populations. Km*3 in Koreans was very similar to those observed in northern Chinese, Japanese, and the native people of British Columbia. It was lower than that reported in Caucasian, Indian, or southern Chinese. PMID- 9339891 TI - The effect of dietary threonine on Adh expression during the development of Drosophila melanogaster. AB - The alcohol dehydrogenase (ADH) activity and ADH cross reacting material (ADH CRM) were measured and Northern blot analysis was carried to define the function and the regulation mechanism of the Adh gene. This study examined how dietary threonine affects the expression of the Adh gene during development of Drosophila melanogaster. Two wild type strains, one homozygous for Adh(F) and one for Adh(S) from Chunan, Korea were used. The ADH activity and CRMs of the Adh(F) strain were 2.1 times higher than those of Adhs strain, and ADH activity was higher with isopropanol (secondary alcohol) than with ethanol (primary alcohol) as a substrate in both Adh(F) and Adh(S) strains. When the larvae, pupae, newly emerged adults (0-1 day), and adults (5-7 days) of Drosophila melanogaster Adh(F) and Adh(S) strains were fed on a defined low yeast and threonine medium, ADH activity and ADH CRM levels were increased. Northern blot analyses indicated that the production of mRNA of the larvae, young adults (0-1 day), and adults (5-7 days) was increased by dietary threonine. ADH activity and ADH CRM increases in Drosophila melanogaster fed on threonine were as a result of threonine-stimulated alteration in the amount of ADH mRNA. The elevated level of the ADH mRNA transcribed from the proximal and distal promoters of threonine-fed larvae and adults showed that there was an induction. PMID- 9339892 TI - Inducible expression of yeast mitochondrial citrate synthase in Aspergillus nidulans. AB - The coding region of the mitochondrial citrate synthase gene (CIT1) from Saccharomyces cerevisiae was amplified by PCR and cloned into an expression vector (pAL4) downstream of the alcohol dehydrogenase (alcA) promoter of Aspergillus nidulans to yield pALCS1. Transformation of A. nidulans A773 with this construct gave stable transformants, AYC#1 and AYC#2, that were phenotypically stable for several mitotic divisions. Southern blot analysis showed that the CIT1 gene was successfully integrated into the chromosomes of the transformants. Western blot analysis and enzymatic assay for citrate synthase revealed that the integrated yeast gene was subject to inducible expression controlled by alcA promoter, which can be induced by threonine. PMID- 9339893 TI - Comparison of various expression plasmids for the induction of immune response by DNA immunization. AB - Intramuscular injection of plasmid DNA is an efficient method to introduce a foreign gene into a live animal. We investigated several factors affecting the gene transfer efficiency and the following immune response by intramuscular injection of plasmid DNA. When the strength of several highly efficient viral promoters was compared in muscle by using the chloramphenicol acetyltransferase (CAT) gene as an indicator, cytomegalovirus (CMV) immediate early promoter was found to be stronger than any other viral promoters including Rous sarcoma virus (RSV), murine leukemia virus (SL3-3) and simian virus 40 (SV40) early promoters. Inclusion of adenovirus tripartite leader (TPL) sequences and a synthetic intron in the 5' untranslated region of mRNA moderately stimulated the CAT expression. On the other hand, the expression of encephalomyocarditis virus (EMCV) VP1 gene was greatly enhanced by the TPL sequences and an intron. The level of humoral immune response by intramuscular injection of various VP1 expression plasmids was compared. The seroconversion rate was highly dependent on the strength of the expression vector. However, the ratio of IgG1 and IgG2a immune response was not significantly variable depending on the strength of the expression vector. Also, the efficiency of the sindbis virus-based DNA vector was examined for the gene expression and immune response. Although a high level of CAT expression was obtained in muscle by using this system, VP1 was not produced as much as the conventional expression vectors. Furthermore, little humoral immune response was elicited by intramuscular injection of VP1-expressing sindbis vector, suggesting that this system was not superior to the conventional vector for DNA immunization. PMID- 9339894 TI - Dicistronic tagging of genes active in embryonic stem cells of mice. AB - A gene-trap vector, pWH14, has been developed to tag genes expressed in embryonic stem (ES) cells of the mouse. The approach relies on the ability of the endogenous promoter to drive promoterless neo-IRES-lacZ construct producing a dicistronic mRNA consisting of the neomycin-resistance (neo) gene and the beta galactosidase gene sequence. The neo gene produces a chimeric protein with the truncated product of the tagged gene and serves as a selectable marker for an insertion into an expressed gene. The internal ribosome entry site (IRES) sequence from murine encephalomyocarditis virus allows the translation of the second cistron, lacZ, to produce beta-galactosidase that can be used as a reporter for the expression of the tagged gene. The pWH14 vector was introduced into ES cells by electroporation, and the cells were selected for G418 resistance. About 50% of the G418-resistant colonies were stained positive for the beta-galactosidase activity. Southern analysis showed that each clone had one or more vector sequences integrated. Northern blot analysis of the clones positive for beta-galactosidase indicated that the fused RNAs containing the neo and the beta-gal genes were derived from the endogenous promoters of the tagged genes. Seven clones were chosen and injected into blastocysts, and chimeras were obtained. Two of the gene-trap insertions (wh14.1 and wh14.3) were transmitted through germ-line. In these two lines, the pattern of lacZ expression was restricted to early stages of embryos. This gene-trap vector may provide a means for tagging and studying the active genes in vivo in early embryogenesis. PMID- 9339895 TI - Hinnavin I, an antibacterial peptide from cabbage butterfly, Artogeia rapae. AB - We have previously isolated four antibacterial peptides from the immune haemolymph of the fifth instar larvae of cabbage butterfly, Artogeia rapae [Yoe, S. M., Bang, I. S., Kang, C. S., and Kim, H. J. (1996) Mol. Cells 6, 609-614]. They were induced by live, nonpathogenic gram negative bacteria. One of these novel antibacterial peptides was named hinnavin I. Hinnavin I is heat stable; its activity was retained after 60 min incubation at 100 degrees C, being effective against gram negative and/or gram positive bacteria. Hinnavin I and lysozyme II showed a powerful synergistic effect on the inhibition of bacterial growth. Amino acid composition was analyzed and the molecular weight was determined to be 4,139.94+/-10.91 Da by the ESI mass spectrometer. To elucidate the primary structure of hinnavin I, the amino acid sequence of this peptide was determined by N-terminal sequencing techniques. The amino-terminal half of the molecule was rich in charged amino acids and was hydrophilic, whereas the carboxyl-terminal half was hydrophobic. PMID- 9339896 TI - Tetracycline-mediated suppression of gene expression with a new dicistronic retroviral vector. AB - Since the first description of the helper-free retrovirus vector by Mann et al. (1983), many improvements have been introduced to the system to increase titer, or to achieve better expression of the transduced genes in cells of different lineage. The typical form of recombinant retrovirus vector utilizes its 5' long terminal repeat (LTR) as a promoter unit for the transcription of the inserted gene(s), which allows only the constitutive expression of the inserted gene(s) in target cells. Although constitutive expression of the delivered gene(s) in target cells may be sufficient for some purposes, controlled expression of gene(s) in target cells or tissues would be favorable in many basic and clinical applications. To circumvent these problems, we developed a new retroviral vector that allows controlled expression of the inserted genes. With these new retroviral vectors, the expression level of the inserted genes can be controlled up to 200-fold in the presence of a minimum amount of tetracycline. We think these new retroviral vectors will be useful in regulating the expression of the genes delivered in vivo. PMID- 9339897 TI - Genetic variation of the apolipoprotein B gene in Korean patients with coronary artery disease. AB - In view of the clinical importance of apolipoprotein B (apo B) as a major risk factor for coronary artery disease (CAD), we investigated the two polymorphisms (signal peptide and XbaI) of the apo B gene in 235 Korean patients with CAD and in 216 normal controls. The insertion allele (INS) frequency of signal peptide polymorphism was significantly higher in cases than the controls (P<0.05). Signal peptide polymorphism was also associated with levels of plasma cholesterol, triglyceride and low density lipoprotein-cholesterol. However, XbaI polymorphism was not associated with plasma lipid levels. Therefore, our results suggest that signal peptide polymorphism of the apo B gene might be one of the factors in explaining an association in Korean CAD patients. PMID- 9339898 TI - Characterization of four yeast artificial chromosome clones mapped to human chromosome 21q22.1 with eight markers. AB - Yeast artificial chromosome (YAC) clones have been successfully utilized to generate a YAC contig map of the long arm of human chromosome 21 (Hu21q). The chromosome subband of 21q22.1 where five genetic loci (IFNAR1, IFNAR2, CRFB4, AF 1, and GART) are mapped is a gene-rich region and needs to be characterized in further detail. YAC D142H8 and YAC F136C5, which were characterized previously by a functional YAC expression procedure, and two new YAC clones, YAC 872B5 and YAC 876D4 located at 21q22.1 whose YAC sizes are 800 kb and 1,500 kb, respectively, were used in this study. To obtain more markers useful for making a detailed physical map of the region, a purified yeast artificial chromosome (YAC D142H8) was used to screen the 3 x 1 S cDNA library. As a result three anonymous cDNA clones (Kmy1, Kmy2, and Qorf4) were obtained, and the nucleotide sequences of Kmy1 and Kmy2 were determined. In an attempt to make a detailed physical map of the region, the locations of five known genes as well as the three new markers were determined on the four YACs by Southern blot analysis. YAC 872B5 contained all markers except GART while YAC F136C5, YAC D142H8, and YAC 876D4 contained three markers (CRFB4, IFNAR1, and IFNAR2), four markers (Kmy1, Kmy2, Qorf4 and AF 1), and four markers (Kmy1, Kmy2, Qorf4 and GART), respectively. YAC 872B5 may represent 1,500 kb of the 21q22.1 subband and half of the 3 x 1 S region, so it should be very useful for studying the relevent region of the human chromosome functionally and physically. PMID- 9339899 TI - Molecular cloning and characterization of the Saccharomyces cerevisiae SAB1 gene that suppresses a temperature-sensitive phenotype of the ARS-binding factor 1 mutant. AB - A high-copy number suppressor gene of the yeast temperature-sensitive lethal abf1 mutant was isolated and named SAB1 (suppressor of ABF1). Chromoblot hybridization and grid-filter hybridization analyses showed that the SAB1 gene was located on chromosome IV. Deletion analyses of the SAB1 plasmid revealed that the suppressor activity was contained in a 1.1 kb DNA region. The nucleotide sequence of the 1.1 kb DNA fragment was determined and turned out to be identical to that of the yeast phosphoribosylanthranilate isomerase gene (TRP1). A binding site for ARS Binding Factor 1 was located in the coding sequence of the TRP1 gene, which has been known to be a part of the B domain of yeast autonomously replicating sequence 1 (ARS1). Our results suggest that ABF1 might be important for the transcription of the yeast TRP1 gene in addition to having important roles in the stimulation of replication at the ARS1 locus. PMID- 9339901 TI - Diversity among clinical isolates of Helicobacter pylori in Korea. AB - Eleven strains of Helicobacter pylori were isolated from biopsy specimens obtained at the A-San Medical Center from December, 1995 to February, 1996. Every H. pylori positive patient was diagnosed to have chronic, erosive, or mild superficial gastritis. To determine the diversity in clinical isolates, the following were studied: total protein profile, plasmid profile, presence of cagA, and variation in DNA sequence. Protein profiles of nine isolates were similar to each other while two isolates had a 35 kDa protein which did not appear in others. The presence of cagA was detected with PCR in seven isolates (63%). Among eleven isolates, seven (63%) carried plasmids. Each isolate showed a big diversity with a PCR-based randomly amplified polymorphic DNA method. Even though all H. pylori isolates used in this study were isolated from gastritis patients at the same hospital, their molecular and biological characteristics were quite different from another showing a big diversity in H. pylori. PMID- 9339902 TI - Molecular characterization of a nonsuppressible allele (prC4) of the Drosophila purple gene. AB - Purple gene encodes 6-pyruvoyl tetrahydropterin synthase (PTP synthase) in Drosophila. The enzyme PTP synthase catalyzes the conversion of dihydroneopterin triphosphate (H2-NTP) to 6-pyruvoyl tetrahydropterin (PTP), an important intermediate for pterin compounds. The extreme purple mutant, prC4, shows a very low activity of PTP synthase. The mutant purple gene has been cloned by screening with the subgenomic library of prC4. The size and expression level of PTP synthase gene transcripts in prC4 were almost the same as those of the wild type. The genomic DNA was also examined in the purple region by Southern blot analysis, but no changes in restriction pattern could be detected. Compared with the wild type PTP synthase sequence, the mutant PTP synthase of prC4 showed three missense mutations: the replacement of alanine 7 by serine (A7S), leucine 9 by phenylalanine (L9P), and aspartic acid 168 by glycine (D168G). Significance of these mutations was discussed in relation to the formation of the oligomeric structure of PTP synthase. PMID- 9339900 TI - Characterization of the murine cyclin D2 gene: exon/intron organization and promoter activity. AB - Cyclin D2 is normally expressed in G1 and promotes progression through G1 of the cell cycle. From a murine genomic library constructed with spleen DNA, two overlapping genomic clones of cyclin D2 were isolated. These clones contain most of the exon of cyclin D2 except exon 5. Characterization of these clones revealed that murine cyclin D2 mRNA spans over 18 kb and 5 exons ranging from 149 to approximately 462 bp in length, and suggested that exon 5 may be at least >5 kb downstream from exon 4. Primer extension analysis of cyclin D2 mRNA isolated from murine activated T cells detected 5 putative sites of transcription initiation. These are located at - 499, - 417, - 391, - 373, and - 349 relative to the translation start site, which is given as + 1. No consensus sequence for TATA box existed at an appropriate position within the promotor region. Instead, several putative transcriptional factor binding sites for C/EBP, PEA3, AP2, NF-Y, Sp1, c Myc, GATA-1, AP1, v-Myb, and CREB were detected. The 5'-flanking region of the cyclin D2 gene up to nucleotide - 945 shared about 61% sequence homology between mouse and human. Functional analysis of promoter activity of the 5'-flanking region of cyclin D2 suggested that the region - 1,100 to - 805 including C/EBP, PEA3, AP2, NF-Y, c-Myc, and Sp1 may have a major positive regulatory activity for expression of cyclin D2. PMID- 9339903 TI - Fragmentation of human Cu,Zn-superoxide dismutase by peroxidative reaction. AB - We investigated the fragmentation of human Cu,Zn-superoxide dismutase (SOD) by H2O2. When Cu,Zn-SOD was incubated with H2O2, the fragmentation of protein proceeded rapidly within 1 min. The amounts of .OH radical formed in the Cu,Zn SOD/H2O2 system reached a maximum in about 3 min. This result suggested that .OH was implicated in the fragmentation step. Copper ions released from Cu,Zn-SOD were gradually increased up to 30 min after starting the incubation with H2O2 in a time-dependent manner. However, the fragmentation was not inhibited by 5 mM DTPA. The results suggested that the fragmentation of Cu,Zn-SOD by H2O2 was due to the peroxidative reaction of SOD rather than the Fenton-like reaction by free copper released from oxidatively damaged SOD. A radical scavenger, azide anion, inhibited the fragmentation of Cu,Zn-SOD and the formation of .OH whereas ethanol did not. These results indicated that azide anions had easy access inside the active channel of Cu,Zn-SOD and thus protected the damage of the enzyme by .OH radicals whereas neutral alcohols stayed outside the active channel and could hardly intercept the newly-formed .OH radicals. Thus we conclude that the fragmentation of Cu,Zn-SOD by H2O2, is due to the oxidative damage resulting from free .OH radicals generated by the peroxidative reaction of SOD. PMID- 9339904 TI - Characterization of two rice MADS box genes that control flowering time. AB - Plants contain a variety of the MADS box genes that encode regulatory proteins and play important roles in both the formation of flower meristem and the determination of floral organ identity. We have characterized two flower-specific cDNAs from rice, designated OsMADS7 and OsMADS8. The cDNAs displayed the structure of a typical plant MADS box gene, which consists of the MADS domain, I region, K domain, and C-terminal region. These genes were classified as members of the AGL2 gene family based on sequence homology. The OsMADS7 and 8 proteins were most homologous to OM1 and FBP2, respectively. The OsMADS7 and 8 transcripts were detectable primarily in carpels and also weakly in anthers. During flower development, the OsMADS genes started to express at the young flower stage and the expression continued to the late stage of flower development. The OsMADS7 and 8 genes were mapped on the long arms of the chromosome 8 and 9, respectively. To study the functions of the genes, the cDNA clones were expressed ectopically using the CaMV 35S promoter in a heterologous tobacco plant system. Transgenic plants expressing the OsMADS genes exhibited the phenotype of early flowering and dwarfism. The strength of the phenotypes was proportional to the levels of transgene expression and the phenotypes were co-inherited with the kanamycin resistant gene to the next generation. These results indicate that OsMADS7 and 8 are structurally related to the AGL2 family and are involved in controlling flowering time. PMID- 9339905 TI - Isolation of molecular markers for salt stress responses in Arabidopsis thaliana. AB - Characterization of many osmotic stress-induced genes has greatly contributed to the understanding of the physiological responses of plant cells to osmotic stress at the molecular level. In this study we constructed a subtraction library and generated 15 salt stress-inducible ESTs from this library to use as molecular markers that reflect the cellular responses to salt stress responses in Arabidopsis. The sequence analysis showed that 5 salt stress-inducible ESTs were identical to previously identified genes in Arabidopsis, 6 cDNAs were homologous to known genes found in plants as well as yeast, and 4 cDNAs were new genes. To confirm that expression of these clones are induced by salt stress, we carried out Northern blot analysis. When we examined for 15 cDNA clones, they were indeed induced by NaCl treatment. The induction level was variable among these genes ranging from approximately 2-fold to more than 50-fold. Also, Northern blot analysis revealed that these genes can be divided into three different induction patterns: early induction, late induction, and continuous induction. PMID- 9339906 TI - Effect of silkworm hemolymph on the expression of E. coli beta-galactosidase in insect cell lines infected with recombinant baculoviruses. AB - The effects of silkworm hemolymph on the expression of foreign genes by recombinant baculoviruses in cell lines were studied. The expression efficiency of beta-galactosidase by recombinant virus containing the E. coli lacZ gene at various concentrations of hemolymph and FBS was determined in BmN and Sf cell lines. The addition of hemolymph to the medium containing FBS accelerated the expression of beta-galactosidase by recombinant viruses in both cells. It was more effective in BmN cells than in Sf cells. Hemolymph was most effective in enhancing virus multiplicity under conditions of 5% FBS. PMID- 9339928 TI - Science, specialization, and the American Surgical Association. PMID- 9339929 TI - Routine pulmonary artery catheterization does not reduce morbidity and mortality of elective vascular surgery: results of a prospective, randomized trial. AB - OBJECTIVE: The authors determined whether the preoperative placement of a pulmonary artery catheter (PAC) with optimization of hemodynamics results in outcome improvement after elective vascular surgery. SUMMARY BACKGROUND DATA: The PAC commonly is used not only in patients who are critically ill, but also perioperatively in major elective surgery. Few prospective studies exist documenting its usefulness. METHODS: One hundred four consecutive patients were randomized to have a PAC placed the morning of operation (group I) or to have a PAC placed only if clinically indicated (group II). Group I patients were resuscitated to preestablished endpoints before surgery and kept at these points both intraoperatively and postoperatively. Group II patients received standard care. RESULTS: There was one death in each group. An intraoperative or postoperative complication developed in 13 patients in group I versus 7 patients in group II (p = not significant). Group I patients received more fluid than did group II patients (5137 +/- 315 mL vs. 3789 +/- 306 mL; p < 0.003). There was no significant difference in either overall or surgical intensive care unit length of stay. Only one patient in group II required a postoperative PAC. CONCLUSIONS: Routine PAC use in elective vascular surgery increases the volume of fluid given to patients without demonstrable improvement in morbidity or mortality. PMID- 9339930 TI - Laparoscopic adrenalectomy: lessons learned from 100 consecutive procedures. AB - One hundred consecutive laparoscopic adrenal procedures for a variety of endocrine disorders were reviewed. There was no mortality, morbidity was 12%, and conversions was 3%. During follow-up, none had recurrence of hormonal excess. Laparoscopic adrenalectomy is the procedure of choice for adrenal removal except in carcinoma or masses > 15 cm. OBJECTIVE: The authors evaluate the effectiveness of laparoscopic adrenalectomy for a variety of endocrine disorders. SUMMARY BACKGROUND DATA: Since the first laparoscopic adrenalectomy was performed in 1992, this approach quickly has been adopted, and increasing numbers are being reported. However, the follow-up period has been too short to evaluate the completeness of these operations. METHODS: One hundred consecutive laparoscopic adrenal procedures from January 1992 until November 1996 were reviewed and followed for adequacy of resection. RESULTS: Eighty-eight patients underwent 97 adrenalectomies and biopsies. The mean age was 46 years (range, 17-84 years). Indications were pheochromocytomas (n = 25), aldosterone-producing adenomas (n = 21), nonfunctional adenomas (n = 20), cortisol-producing adenomas (n = 13), Cushing's disease (n = 8), and others (n = 13). Fifty-five patients had previous abdominal surgery. Mean operative time was 123 minutes (range, 80-360 minutes), and estimated blood loss was 70 mL (range, 20-1300 mL). There was no mortality, and morbidity was encountered in 12% of patients, including three patients in whom venous thrombosis developed with two sustaining pulmonary emboli. During pheochromocytoma removal, hypertension occurred in 56% of patients and hypotension in 52%. There were three conversions to open surgery. The average length of stay has decreased from 3 days (range, 2-19 days) in the first 3 years to 2.4 days (range, 1-6 days) over the past 16 months. During follow-up (range, 1 44 months), two patients had renovascular hypertension and none had recurrence of hormonal excess. CONCLUSION: Laparoscopic adrenalectomy is safe, effective, and decreases hospital stay and wound complications. Prior abdominal surgery is not a contraindication. Pheochromocytomas can be resected safely laparoscopically despite blood pressure variations. Venous thrombosis prophylaxis is mandatory. The laparoscopic approach is the procedure of choice for adrenalectomy except in the case of invasive carcinoma or masses > 15 cm. PMID- 9339931 TI - Six hundred fifty consecutive pancreaticoduodenectomies in the 1990s: pathology, complications, and outcomes. AB - OBJECTIVE: The authors reviewed the pathology, complications, and outcomes in a consecutive group of 650 patients undergoing pancreaticoduodenectomy in the 1990s. SUMMARY BACKGROUND DATA: Pancreaticoduodenectomy has been used increasingly in recent years to resect a variety of malignant and benign diseases of the pancreas and periampullary region. METHODS: Between January 1990 and July 1996, inclusive, 650 patients underwent pancreaticoduodenal resection at The Johns Hopkins Hospital. Data were recorded prospectively on all patients. All pathology specimens were reviewed and categorized. Statistical analyses were performed using both univariate and multivariate models. RESULTS: The patients had a mean age of 63 +/- 12.8 years, with 54% male and 91% white. The number of resections per year rose from 60 in 1990 to 161 in 1995. Pathologic examination results showed pancreatic cancer (n = 282; 43%), ampullary cancer (n = 70; 11%), distal common bile duct cancer (n = 65; 10%), duodenal cancer (n = 26; 4%), chronic pancreatitis (n = 71; 11%), neuroendocrine tumor (n = 31; 5%), periampullary adenoma (n = 21; 3%), cystadenocarcinoma (n = 14; 2%), cystadenoma (n = 25; 4%), and other (n = 45; 7%). The surgical procedure involved pylorus preservation in 82%, partial pancreatectomy in 95%, and portal or superior mesenteric venous resection in 4%. Pancreatic-enteric reconstruction, when appropriate, was via pancreaticojejunostomy in 71% and pancreaticogastrostomy in 29%. The median intraoperative blood loss was 625 mL, median units of red cells transfused was zero, and the median operative time was 7 hours. During this period, 190 consecutive pancreaticoduodenectomies were performed without a mortality. Nine deaths occurred in-hospital or within 30 days of operation (1.4% operative mortality). The postoperative complication rate was 41%, with the most common complications being early delayed gastric emptying (19%), pancreatic fistula (14%), and wound infection (10%). Twenty-three patients required reoperation in the immediate postoperative period (3.5%), most commonly for bleeding, abscess, or dehiscence. The median postoperative length of stay was 13 days. A multivariate analysis of the 443 patients with periampullary adenocarcinoma indicated that the most powerful independent predictors favoring long-term survival included a pathologic diagnosis of duodenal adenocarcinoma, tumor diameter <3 cm, negative resection margins, absence of lymph node metastases, well-differentiated histology, and no reoperation. CONCLUSIONS: This single institution, high-volume experience indicates that pancreaticoduodenectomy can be performed safely for a variety of malignant and benign disorders of the pancreas and periampullary region. Overall survival is determined largely by the pathology within the resection specimen. PMID- 9339932 TI - Adult-to-adult living donor liver transplantation using extended right lobe grafts. AB - OBJECTIVE: The authors report their experience with living donor liver transplantation (LDLT) using extended right lobe grafts for adult patients under high-urgency situations. SUMMARY BACKGROUND DATA: The efficacy of LDLT in the treatment of children has been established. The major limitation of adult-to adult LDLT is the adequacy of the graft size. A left lobe graft from a relatively small volunteer donor will not meet the metabolic demand of a larger recipient. METHODS: From May 1996 to November 1996, seven LDLTs, using extended right lobe grafts, were performed under high-urgency situations. All recipients were in intensive care units before transplantation with five having acute renal failure, three on mechanical ventilation, and all with hepatic encephalopathy. The median body weight for the donors and recipients was 58 kg (range, 41-84 kg) and 65 kg (range, 53-90 kg), respectively. The body weights of four donors were less than those of the corresponding recipients, and the lowest donor-to-recipient body weight ratio was 0.62:1. The extended right lobe graft was chosen because the left lobe volume was <40% of the ideal liver mass of the recipient. RESULTS: Median blood loss for the donors was 900 mL (range, 700-1600 mL) and hospital stay was 19 days (range, 8-22 days). Homologous blood transfusion was not required. Two donors had complications (one incisional hernia and one bile duct stricture) requiring reoperation after discharge. All were well with normal liver function 5 to 10 months after surgery. The graft weight ranged from 490 g to 1140 g. All grafts showed immediate function with normalization of prothrombin time and recovery of conscious state of the recipients. There was no vascular complication, but six recipients required reoperation. One recipient died of systemic candidiasis 16 days after transplantation and 6 (86%) were alive with the original graft at a median follow-up of 6.5 months (range, 5-10 months). CONCLUSIONS: When performed by a team with experience in hepatectomy and transplantation, LDLT, using an extended right lobe graft, can achieve superior results. The technique extends the success of LDLT from pediatric recipients to adult recipients and opens a new donor pool for adults to receive a timely graft of adequate function. PMID- 9339933 TI - Histopathologic validation of the sentinel lymph node hypothesis for breast carcinoma. AB - BACKGROUND AND OBJECTIVE: The sentinel node hypothesis assumes that a primary tumor drains to a specific lymph node in the regional lymphatic basin. To determine whether the sentinel node is indeed the node most likely to harbor an axillary metastasis from breast carcinoma, the authors used cytokeratin immunohistochemical staining (IHC) to examine both sentinel and nonsentinel lymph nodes. METHODS: From February 1994 through October 1995, patients with breast cancer were staged with sentinel lymphadenectomy followed by completion level I and II axillary dissection. If the sentinel node was free of metastasis by hematoxylin and eosin staining (H&E), then sentinel and nonsentinel nodes were examined with IHC. RESULTS: The 103 patients had a median age of 55 years and a median tumor size of 1.8 cm (58.3% T1, 39.8% T2, and 1.9% T3). A mean of 2 sentinel (range, 1-8) and 18.9 nonsentinel (range, 7-37) nodes were excised per patient. The H&E identified 33 patients (32%) with a sentinel lymph node metastasis and 70 patients (68%) with tumor-free sentinel nodes. Applying IHC to the 157 tumor-free sentinel nodes in these 70 patients showed an additional 10 tumor-involved nodes, each in a different patient. Thus, 10 (14.3%) of 70 patients who were tumor-free by H&E actually were sentinel node-positive, and the IHC lymph node conversion rate from sentinel node-negative to sentinel node positive was 6.4% (10/157). Overall, sentinel node metastases were detected in 43 (41.8%) of 103 patients. In the 60 patients whose sentinel nodes were metastasis free by H&E and IHC, 1087 nonsentinel nodes were examined at 2 levels by IHC and only 1 additional tumor-positive lymph node was identified. Therefore, one H&E sentinel node-negative patient (1.7%) was actually node-positive (p < 0.0001), and the nonsentinel IHC lymph node conversion rate was 0.09% (1/1087; p < 0.0001). CONCLUSIONS: If the sentinel node is tumor-free by both H&E and IHC, then the probability of nonsentinel node involvement is <0.1%. The true false negative rate of this technique using multiple sections and IHC to examine all nonsentinel nodes for metastasis is 0.97% (1/103) in the authors' hands. The sentinel lymph node is indeed the most likely axillary node to harbor metastatic breast carcinoma. PMID- 9339935 TI - Factors predicting a successful outcome after pharmacologic bowel compensation. AB - OBJECTIVES: The authors determined those factors that predict a successful outcome in patients who receive pharmacologic agents to promote bowel absorption after massive intestinal resection. SUMMARY BACKGROUND DATA: Patients with the short bowel syndrome are maintained on long-term total parenteral nutrition (TPN) or more frequently considered for intestinal transplantation as part of their treatment program. The authors have administered a combination of trophic agents and a specialized diet to further enhance intestinal compensation and optimize nutrient absorption in patients with intestinal failure. METHODS: Forty-five TPN dependent adults with a jejunal-ileal remnant < or = 50 cm and a portion of colon in continuity were treated with growth hormone, glutamine, and a modified diet for 4 weeks and observed for an average of 1.8 years. RESULTS: The average age of the patients was 43 years, the average jejunal-ileal length was 23 cm, and the average length of time the patient received TPN was 4.3 years. After 4 weeks of therapy, 26 (58%) were free of TPN support. Predictors of a favorable response included greater bowel length, lower body weight, and greater bowel length-body weight ratio. At follow-up, the percentage of patients who were not receiving TPN had fallen to 40%. CONCLUSIONS: Approximately half of a group of patients, thought to have absorptive surface area inadequate to be independent of TPN support, can maintain themselves on enteral feedings after this intestinal rehabilitation program. Because of the risk, costs, and alterations in lifestyle associated with long-term TPN or intestinal transplantation or both, it seems prudent to consider a program of bowel rehabilitation with an individual patient before embarking on another therapeutic plan. PMID- 9339934 TI - Does information from axillary dissection change treatment in clinically node negative patients with breast cancer? An algorithm for assessment of impact of axillary dissection. AB - OBJECTIVE: The authors assessed the impact of axillary dissection on adjuvant systemic therapy recommendations in patients with breast cancer. SUMMARY BACKGROUND DATA: With increasing use of systemic therapy in node-negative women and the desire to reduce treatment morbidity and cost, the need for axillary dissection in clinically node-negative patients with breast cancer has been challenged. METHODS: Two hundred eighty-two women with clinically negative axillae were analyzed using a model treatment algorithm. Systemic therapy was assigned with and without data from axillary dissection. Treatment shifts based on axillary dissection data were scored. RESULTS: Twenty-seven percent of clinically node-negative women had pathologically positive nodes. Eight percent of T1a and 10% of T1b tumors had positive nodes and would have been undertreated without axillary dissection. Seven percent of premenopausal women with tumors < 1 cm and 13% with tumors > or = 1 cm had treatment changed by axillary dissection. For women 50 to 60 years of age, 10% with tumors < 1 cm, 17% with tumors 1 to 2 cm with positive prognostic features, and 4% with poor prognostic features had significant treatment shifts after axillary dissection. For clinically node negative women older than 60 years of age not eligible for chemotherapy, only 3% of those with tumors < 1 cm and none of those with tumors > or = 1 cm had their treatment changed by findings at axillary dissection. Treatment shifts based on axillary dissection were larger if the treatment algorithm allowed for more varied or more aggressive treatment options. CONCLUSIONS: Data obtained from axillary dissection will alter adjuvant systemic therapy regimen in a significant number of clinically node-negative women younger than 60 years of age and for older women eligible to receive chemotherapy. PMID- 9339936 TI - Thoracoabdominal aneurysm repair: perspectives over a decade with the clamp-and sew technique. AB - OBJECTIVES: Experience over a decade with thoracoabdominal aortic aneurysm (TAA) repair using a clamp-sew technique was reviewed to compare overall results with alternative operative methods. SUMMARY BACKGROUND DATA: Controversy continues as to the optimal technique for TAA repair, with frequent contemporary emphasis on bypass-distal perfusion methods. Proponents of this technique claim improved results compared to those of historic control subjects in the parameters of operative mortality, postoperative renal failure, and lower extremity neurologic deficit. METHODS: Over the interval from 1987 to 1996, 160 TAA repairs (type I, 32%; type II, 15%; type III, 34%; and type IV, 19%) were performed in 157 patients with a mean age of 70 years and a male-to-female ratio of 1/1. Clinical features included ruptured TAA (10%), urgent operation (22.5%), and aortic dissection (18%). Operative management used a clamp-sew technique with regional hypothermia for spinal cord (epidural cooling, since 1993) and renal protection. Variables associated with the endpoints of operative mortality or major morbidity, particularly spinal cord injury, were assessed with Fisher exact test and logistic regression; late survival was estimated with the Kaplan-Meier method. RESULTS: In-hospital mortality was 9% and was associated with operation for rupture (p < 0.005) or other acute presentation (p < 0.001). After multivariate analysis, the postoperative complication renal failure (relative risk, 6.5 [95% confidence interval, 1.8-23.6, p = 0.004]) and significant spinal cord injury (relative risk, 16.5 [95% confidence interval, 3.2-83.2, p = 0.001]) were associated independently with operative mortality. Paraparesis-paraplegia occurred in 7%, an incidence significantly (p < 0.001) less than that (18.7%) predicted for this cohort from published models. Variables associated (univariate analysis) with this complication included TAA rupture (p < 0.0001), other acute presentation or dissection (p < 0.001), prolonged (>6 hours) operation (p < 0.04), and excessive (>3 L) transfusions (p < 0.02). Operation for acute presentation or dissection (relative risk, 7.9 [95% confidence interval, 1.7 37.7, p = 0.009]) and prolonged surgery [relative risk, 7.5 [95% confidence interval, 1.5-35.3, p = 0.01]) retained independent association with paraplegia paraparesis after multivariate analysis. Dialysis was needed in 2.5%. Late survival at 1 and 5 years was 86 +/- 2.9% and 62 +/- 5.8%, respectively. CONCLUSIONS: These data compare favorably with those from contemporary reports using other operative strategies and do not support routine adoption of bypass distal perfusion as the preferred technique for TAA repair. PMID- 9339937 TI - The fibrinolytic effects of intermittent pneumatic compression: mechanism of enhanced fibrinolysis. AB - BACKGROUND AND OBJECTIVES: Intermittent pneumatic compression (IPC) is an effective form of deep vein thrombosis prophylaxis for general surgery patients. The antithrombotic effect of IPC is thought to be the result of increased venous velocity and stimulation of endogenous fibrinolysis. However, the mechanism of enhanced fibrinolytic activity and the relative effects on normal and postthrombotic veins have not been defined. The purposes of this study are 1) to quantify changes in fibrinolytic activity with IPC; 2) to study the mechanism of fibrinolytic enhancement with IPC; and 3) to evaluate whether postthrombotic patients have the same capacity for fibrinolytic enhancement with IPC as do normal subjects. METHODS: Twelve volunteers (6 normal and 6 postthrombotic) had 5 IPC devices applied for 120 minutes in random fashion, 1 per week x 5 weeks. The devices included single-chamber, sequential, foot, calf, and long-leg compression. Subjects had an indwelling antecubital venous cannula placed for blood drawn at baseline, 60, 120, and 180 minutes after IPC devices were applied. Global fibrinolytic activity (euglobulin fraction, fibrin plate assay), tissue plasminogen activator (tPA) antigen (Ag) and activity (Act), plasminogen activator inhibitor-1 (PAI-1) Ag and Act, alpha-2-antiplasmin-plasmin complexes, and von Willebrand factor (vWF) antigen were assayed. RESULTS: A striking elevation in fibrinolytic activity was noted at 180 minutes with all devices in normal subjects and postthrombotic patients (p = 0.01-0.0001); however, baseline and stimulated fibrinolytic activity was attenuated in postthrombotic patients (<0.03). The tPA-Act increased only in normal subjects (3.8 +/- 1.9%) (p = 0.057), despite a decrease in plasma tPA-Ag, which was observed in both normal subjects (-12.4 +/- 3.8%) (p = 0.009) and patients (-17.2 +/- 3.1%) (p = 0.001). PAI-1-Ag decreased in both normal subjects (-13.4 +/- 3.8%) (p = 0.007) and patients (-12.0 +/- 3.1%) (p = 0.013) with a marked reduction in PAI-1-Act in both normal subjects (p = 0.003) and patients (p = 0.004). There were no changes in vWF, and alpha-2-antiplasmin-plasmin complexes increased only in postthrombotic patients (p = 0.021). CONCLUSIONS: Stimulation of endogenous fibrinolytic activity occurs after IPC, both in normal subjects and postthrombotic patients; however, baseline and overall fibrinolytic response in postthrombotic patients is reduced. The mechanism of increased fibrinolytic activity is likely because of a reduction in PAI-1, with a resulting increase of tPA activity. PMID- 9339938 TI - Reoperation after Nissen fundoplication in children with gastroesophageal reflux: experience with 130 patients. AB - OBJECTIVE: The authors evaluate reoperation for recurrent gastroesophageal reflux (GER) after a failed Nissen fundoplication. SUMMARY BACKGROUND DATA: Nissen fundoplication is an accepted treatment for GER refractory to medical therapy. Wrap failure and recurrence of GER are noted in 8% to 12%. METHODS: Medical records of 130 children undergoing a second antireflux operation for recurrent GER from January 1985 to June 1996 retrospectively were reviewed. RESULTS: One hundred one patients (78%) were neurologically impaired (NI), 74 (57%) had chronic pulmonary disease, and 8 had esophageal atresia. Recurrent symptoms included vomiting (78%), growth failure (62%), choking-coughing-gagging (38%), and pneumonia (25%). Gastroesophageal reflux was confirmed by barium swallow, gastric scintigraphy, and endoscopy. Operative findings showed wrap breakdown (42%), wrap-hiatal hernia (30%), or both (21%). A second Nissen fundoplication was performed in 128 children. Complications included bowel obstruction (18), wound infection (10), pneumonia (6) and tight wrap (9). There were two postoperative (<30 days) deaths (1.5%). Of 124 patients observed long term, 89 (72%) remain symptom free. Eight were converted to tube feedings. Twenty-seven required a third fundoplication, and 19 (70%) were successful outcome. Two with repetitive wrap failure due to gastric atony underwent gastric resection and esophagojejunostomy. CONCLUSION: Nissen fundoplication was successful in 91% of patients. In 9% with wrap failure, a second Nissen fundoplication was successful in 72%. Reoperation is justified in properly selectedpatients. Conversion to jejunostomy feedings is suggested for neurologically impaired after two wrap failures and a partial wrap in those with esophageal atresia and severe esophageal dysmotility. Repeated wrap failure due to gastric atony requires gastric resection and esophagojejunostomy. PMID- 9339939 TI - Extended cervicomediastinal thymectomy in the integrated management of myasthenia gravis. AB - OBJECTIVE: The authors evaluated the response to extended cervicomediastinal thymectomy as a component of the integrated management of patients with myasthenia gravis in a large series of patients from a single institution. The authors evaluated the response to therapy with respect to a graded, multivariate, ordinal scale chosen to reflect the full range of the disease's manifestations. SUMMARY BACKGROUND DATA: A number of series, of varying sizes, describe the response of patients with myasthenia gravis to thymectomy primarily with respect to the bivariate endpoint of the presence or absence of "remission." These studies fail to describe the full spectrum of postoperative disease severity and have been unable to quantify definitively the influence of putative prognostic variables, nor to assess definitively the statistical significance of apparent improvements over time. METHODS: The authors evaluated 202 consecutive patients who underwent trans-sternal thymectomy for symptomatic myasthenia gravis from 1969 through 1996 at the Johns Hopkins Hospital. The response to surgery, combined with postoperative medical therapy, was evaluated by a standardized scale reflecting the full spectrum of the disease. These data were analyzed by a novel mean multivariate regression analysis, which allowed the quantification of the statistical significance of apparent responses over time and the evaluation of the independent influence of each of nine putative prognostic indicators. RESULTS: There was no perioperative mortality and a 33% perioperative morbidity. There was a marked clinical response at 6 months to 1 year after surgery that was sustained for at least 10 years thereafter. The median increment of improvement was two (of five) classes. Eighty-six percent and 83% of the patients had improved by at least one class at 5 and 10 years, respectively. These changes were statistically significant (p < 0.001). Whereas the use of extended cervicomediastinal thymectomy was associated with a greater than twofold chance of improvement, compared to conventional trans-sternal thymectomy, neither the pathologic diagnosis (presence of thymoma) nor the age at surgery proved to be negative predictors of postoperative response. CONCLUSIONS: Extended cervicomediastinal thymectomy is the procedure of choice as a component of the integrated management of myasthenia gravis, with significant improvement seen in the group as a whole, as well as in subgroups of patients with thymoma and those older than 40 years of age. PMID- 9339940 TI - Differential clinical workloads among faculty at a major academic health center. AB - OBJECTIVE: The authors analyzed patient care (1981-1995) and financial data (1991 1996) to determine if differential workloads existed at a major academic health center. SUMMARY BACKGROUND DATA: Academic health centers differ markedly from community-based medical centers, but they are required to compete with others who have a more circumscribed mission and a responsibility for providing less complex care. Changes in health care systems may lessen incentives to generate clinical revenue and may adversely affect educational and research programs. METHODS: Patient care data at the University of Michigan Health System were analyzed by discipline for level of activity from 1981 to 1995 and were compared to professional and institutional financial data from 1991 to 1995. RESULTS: Surgeons represented 11% of the total full-time physicians throughout the period of the study (94 of the 836 Medical Center physicians, 1995). They accounted for 33% of hospital admissions (11,616 of 35,101) and 16% of outpatient visits (92,364 of 568,738). Since 1981, surgeons experienced a 249% increase in total operative workload (6799-16,909 procedures), representing a 30% increase in operations/surgeon (138-180 operations). Surgical efforts in 1995 accounted for 29% of the total professional fee revenue and $240 million of the $512-million University of Michigan Hospital revenue. CONCLUSIONS: Surgeons had a greater collective and individual responsibility than did nonsurgeons for clinical activity and the financial viability of the academic health centers studied. Many proposals for financing health care delivery systems have the potential to exacerbate this differential. Restructuring of academic health centers must address this fact, lest their academic mission and scholarly activity be compromised. PMID- 9339941 TI - A multivariable risk factor analysis of the portoenterostomy (Kasai) procedure for biliary atresia: twenty-five years of experience from two centers. AB - OBJECTIVE: The authors investigated risk factors for failure after portoenterostomy for biliary atresia using univariate and multivariable methods. SUMMARY BACKGROUND DATA: Kasai's portoenterostomy has gained worldwide acceptance as the initial surgical therapy for infants with biliary atresia. Although extended survival has been achieved for many patients, factors influencing outcome have not been defined clearly. METHODS: The authors analyzed risks for failure in 266 patients treated from 1972 to 1996 by the Kaplan-Meier product limit estimate and Cox proportional hazards model. Failure was defined as death or transplant. RESULTS: Age at surgery, surgical decade, and anatomy of atretic bile ducts were identified as independent risk factors. Five-year survival was 49% and median survival was 15 years when bile drainage was achieved. Sixty-five patients had liver transplants. Mean age at transplant was 5.4 years. CONCLUSIONS: The outcome after portoenterostomy for biliary atresia is determined by age at surgery and anatomy of the atretic extrahepatic bile ducts. Liver transplant will salvage patients with failed Kasai with 10-year posttransplant survival of 71%. PMID- 9339942 TI - Recurrence-free long-term survival after liver transplantation for hepatitis B using interferon-alpha pretransplant and hepatitis B immune globulin posttransplant. AB - OBJECTIVE: The authors determined whether pretransplant reduction of hepatitis B virus (HBV) load using alpha-interferon-2b (IFN) and passive immunoprophylaxis using hepatitis B immunoglobulin (HBIg) posttransplantation can prevent HBV recurrence in patients undergoing liver transplantation (LT) for HBV cirrhosis. SUMMARY BACKGROUND DATA: Liver transplantation in patients with HBV cirrhosis is associated with a high rate of recurrence and reduced survival. In patients with evidence of replicating virus (HBV-DNA or hepatitis B e antigen [HBeAg]-positive serum or both), recurrence is nearly universal. Passive immunoprophylaxis with HBIg alone is not effective in preventing HBV recurrence posttransplant, especially in patients with evidence of active viral replication pretransplant. Higher doses of HBIg posttransplant has reduced recurrence rates to 30% to 50%. Lamivudine, a nucleoside analogue that has shown early promise, also is associated with significant HBV recurrence. The authors report a reliable method of preventing viral recurrence in patients even with evidence for active HBV replication pretransplant. METHODS: Pretransplant patients with evidence of replicating HBV were given IFN starting at 1 million IU 3 times per week subcutaneously. This dose was increased to 2 and then 3 million IU 3 times per week when patient's side effects permitted and was maintained until the patient underwent a LT. All patients were tested every 4 weeks for hepatitis B surface antigen (HBsAg), HBeAg, and HBV-DNA. When patients became negative for HBeAg and HBV-DNA, they were listed for LT. Patients that were only HBsAg positive were listed immediately and received a LT without prior IFN treatment. Post-LT, all patients began receiving HBIg 2000 IU (10 mL) daily from days 1 to 20 and then weekly for the first 2 years. After 2 years, all patients received 2000 IU (10 mL) monthly. Additional HBIg immunoprophylaxis was given during intense immunosuppression for rejection. Posttransplant serum was tested for HBsAg, HBeAg, and HBV-DNA in all patients 1 week, 1 month, and every 3 months thereafter. Liver biopsies were done at least yearly and when liver enzymes were abnormal and were always tested for HBsAg and HBcAg by immunoperoxidase. RESULTS: Thirteen patients with decompensated HBV cirrhosis were transplanted. Pretransplant, eight patients had evidence of active viral replication at the initial assessment (HBeAg or HBV-DNA-positive serum or both). All eight were successfully treated with IFN (median duration, 24 weeks; range, 8-53) and converted to a negative status before transplantation. Side effects from IFN were minimal and well tolerated, except in one patient who required 6 million IU to convert to a nonreplicating status. The five patients that were only HBsAg positive were not treated with IFN pretransplant. After surgery, HBIg given as described achieved consistently serum levels greater than 1000 IU/L. Twelve of the 13 patients are alive with normal liver function and without serologic evidence of HBV recurrence at a median follow-up of 32 months (range, 9-56 months). None have evidence of HBV recurrence as measured by serum HBsAg/HBeAg/HBV-DNA at recent follow-up. The sera of the seven longest survivors has tested negative for HBV-DNA using the polymerase chain reaction method. In addition, a liver biopsy was obtained in six of these patients, the results of which also tested negative for HBV-DNA using polymerase chain reaction. Liver biopsy specimens have been negative for the presence of HBsAg and HBcAg by immunoperoxidase staining in all 12 patients. CONCLUSION: A reduction of viral load pretransplant with IFN and posttransplant HBIg prevents recurrence of hepatitis B and permits LT for HBV cirrhosis, even in patients with evidence of replicating virus. The IFN pretransplant was well tolerated, and the small frequent dosing of HBIg posttransplant did not cause side effects while achieving serum levels > 1000 IU/L. PMID- 9339943 TI - Immediate postoperative enteral feeding results in impaired respiratory mechanics and decreased mobility. AB - OBJECTIVE: The authors set out to determine whether immediate enteral feeding minimizes early postoperative decreases in handgrip and respiratory muscle strength. SUMMARY BACKGROUND DATA: Muscle strength decreases considerably after major surgical procedures. Enteral feeding has been shown to restore strength rapidly in other clinical settings. METHODS: A randomized, controlled, nonblinded clinical trial was conducted in patients undergoing esophagectomy or pancreatoduodenectomy who received immediate postoperative enteral feeding via jejunostomy (fed, n = 13), or no enteral feeding during the first 6 postoperative days (unfed, n = 15). Handgrip strength, vital capacity, forced expiratory volume in one second (FEV1), and maximal inspiratory pressure (MIP) were measured before surgery and on postoperative days 2, 4, and 6. Fatigue and vigor were evaluated before surgery and on postoperative day 6. Mobility was assessed daily after surgery using a standardized descriptive scale. Postoperative urine biochemistry was evaluated in daily 24-hour collections. RESULTS: Postoperative vital capacity (p < 0.05) and FEV1 (p = 0.07) were consistently lower (18%-29%) in the fed group than in the unfed group, whereas grip strength and maximal inspiratory pressure were not significantly different. Postoperative mobility also was lower in the fed patients (p < 0.05) and tended to recover less rapidly (p = 0.07). Fatigue increased and vigor decreased after surgery (both p < or = 0.001), but changes were similar in the fed and unfed groups. Intensive care unit and postoperative hospital stay did not differ between groups. CONCLUSIONS: Immediate postoperative jejunal feeding was associated with impaired respiratory mechanics and postoperative mobility and did not influence the loss of muscle strength or the increase in fatigue, which occurred after major surgery. Immediate postoperative enteral feeding should not be routine in well-nourished patients at low risk of nutrition-related complications. PMID- 9339945 TI - Radiation-induced genomic instability: delayed mutagenic and cytogenetic effects of X rays and alpha particles. AB - The frequency of mutations at the Hprt locus was measured in clonal populations of Chinese hamster ovary cells derived from single cells surviving exposure to 0 12 Gy of X rays or 2 Gy of alpha particles. Approximately 8-9% of 446 clonal populations examined 23 population doublings after irradiation showed high frequencies of late-arising mutations as indicated by mutant fractions 10(2) 10(4)-fold above background. The frequency with which such clones occurred was similar for alpha-particle irradiation and X irradiation, with no apparent dose dependence for X irradiation over the range of 4-12 Gy. The molecular structure of Hprt mutations was determined by analysis by multiplex polymerase chain reaction of all nine exons. Of mutations induced directly after exposure to X rays, 75% involved partial or total gene deletions. Only 19-23% of late-arising (delayed) mutations were associated with deletions, the preponderance of these being partial deletions involving one or two exons. This spectrum was very similar to that for spontaneously arising mutations. To determine whether delayed mutations were non-clonal, the spectrum of exons deleted was examined among 29 mutants with partial deletions derived from a single clonal population. The results indicated that at least 15 of these mutants arose independently. To examine the relationship between the occurrence of delayed mutations and chromosomal instability, 60 Hprt mutant subclones isolated from a clonal population showing a high frequency of delayed mutations were serially cultivated in vitro. Of these, 14 showed a slow-growth phenotype with a high frequency of polyploid cells (10-38%) and a markedly enhanced frequency of non-clonal chromosomal rearrangements including both chromosome-type and chromatid-type aberrations. These clones also showed a 3- to 30-fold increase in the frequency of ouabain-resistant mutations; no ouabain-resistant mutants were induced directly by X irradiation. These results suggest that among clones showing a high frequency of delayed mutations there may be a subpopulation of cells that are particularly unstable; selection for the slow-growth phenotype has the effect of selecting for this chromosomally unstable subpopulation. PMID- 9339944 TI - Modular bifurcation endoprosthesis for treatment of abdominal aortic aneurysms. AB - OBJECTIVE: The authors analyzed a single group's experience treating abdominal aortic aneurysms (AAAs) with a new self-expanding, modular, bifurcated device. SUMMARY BACKGROUND DATA: Successful exclusion of AAAs by prototype devices has led to several controlled clinical trials evaluating prostheses designed and manufactured specifically for this application. METHODS: Sixteen patients (15 males, 1 female) of American Society of Anesthesiologists grade 2 through 4 and average age of 72 years had AAAs (average 57-mm diameter) treated as part of a phase I Food and Drug Administration-approved trial. RESULTS: All patients were treated successfully with no surgical conversions. No endoleaks or aneurysm enlargement was noted either predischarge by contrast computed tomography or on follow-up at 1 month by duplex ultrasound examination. At 6 months, 12 of 13 patients who were observed for this interval had no endoleaks, whereas one patient (patient 3) showed a small area of extravasation that appeared to arise from the device in an area that was traumatized at the time of deployment. One procedure-related mortality (6%) occurred in a patient who died of septic complications secondary to a gangrenous gallbladder diagnosed 1 day after the procedure. There were no device-related mortalities. Complications included two iliac artery dissections, two groin wound infections, and two transient elevations of serum creatinine. Other significant variables including median procedure length (5 hours), intensive care unit stay (1 day), hospitalization postprocedure (4.5 days), and blood loss (1100 mL) all decreased as the study progressed. Blood replacement in all but three patients was accomplished by autotransfusion or banked-autologous blood replacement. At 6-month follow-up in 13 patients, the maximum diameter of the aneurysm decreased by an average of 5.6 mm (range, 0-15 mm), and the maximal cross-sectional area decreased an average of 20.3% (range, 0-72%). CONCLUSIONS: This study suggests that endovascular prosthesis exclusion of AAAs using a self-expanding modular device may be effective in many patients who are otherwise surgical candidates for repair if further clinical studies confirm these observations. PMID- 9339946 TI - Constancy of the relative biological effectiveness of 42 MeV (p-->Be+) neutrons among cell lines with different DNA repair proficiencies. AB - An important approach to understanding the role of the various DNA repair pathways in the cellular response to DNA-damaging agents is through the study of repair-deficient mutant cell lines. In the present study we used this strategy to assess the relative importance of four of these pathways for the repair of DNA damage induced by low-linear energy transfer (LET) gamma rays and intermediate LET 42 MeV (p-->Be+) fast neutrons. The panel of hamster cell mutants that we characterized for their relative sensitivity to fast neutrons and gamma rays includes cell lines with defects in the nucleotide excision repair pathway; these can be further subdivided into mutants which are defective in nucleotide excision repair alone [UV5 (ERCC2-), UV24 (ERCC3-), UV135 (ERCC5-) and UV61 (ERCC6-)] compared to those which have an associated defect in the distinct but overlapping pathway for the repair of DNA crosslinks [UV20 (ERCC1-) and UV41 (ERCC4-)]. We also examined mutants with defects in the base excision repair pathway [EM9 (XRCC1-)] and the DNA-dependent protein kinase (DNA-PK)-mediated DNA double strand break (DSB) repair pathway [xrs5 (XRCC5-)]. None of the mutants defective in nucleotide excision repair was differentially sensitized to fast neutrons or gamma rays; in fact, the slight radiosensitivity of these mutants under aerated conditions may be secondary to their defect in nucleotide excision repair. In contrast, deficiency in the base excision repair pathway resulted in a significant primary sensitization to both types of radiation (1.95-fold to gamma rays and 1.79-fold to neutrons). Deficiency in the DSB repair pathway mediated by DNA-PK resulted in a marked, but again similar, primary sensitization to gamma rays (4.2-fold) and neutrons (5.1-fold). Thus none of the repair pathways examined here exhibited a preferential role for the repair of damage induced by low-LET compared to intermediate-LET radiations; this resulted in an essentially constant relative biological effectiveness (RBE) of approximately 2 among the cell lines studied, independent of their DNA repair proficiency. However, consideration of these data along with data published previously for high-LET alpha particles suggests that, whereas the DNA-PK pathway is important for the repair of DSBs induced by low- and intermediate-LET radiations, it becomes less important as the LET increases beyond 100 keV/microm; thus this pathway may not be involved in repairing the more complex lesions induced by densely ionizing high-LET particles. PMID- 9339947 TI - Induction and rejoining of DNA double-strand breaks in Chinese hamster V79-4 cells irradiated with characteristic aluminum K and copper L ultrasoft X rays. AB - Characteristic aluminum K (AlK) (energy of 1.5 keV) and copper L (CuL) (energy of approximately 0.96 keV) ultrasoft X rays have been used to investigate the effectiveness of the numerous low-energy secondary electrons produced by low linear energy transfer (LET) ionizing radiation. Cellular inactivation and induction and rejoining of DNA double-strand breaks (DSBs) in Chinese hamster V79 4 cells irradiated as monolayers with these ultrasoft X radiations have been studied under aerobic and anaerobic conditions. The mean cell thickness, determined by confocal laser scanning fluorescence microscopy, was used to calculate the mean dose to the nucleus of the irradiated cells. Relative to 60Co gamma rays, the relative biological effectiveness (RBE) for cellular inactivation at 10% survival is 1.7 +/- 0.1 and 2.3 +/- 0.3 for AIK and CuL ultrasoft X rays, respectively. The RBE values for induction of DSBs of 2.5 +/- 0.2 and 3.0 +/- 0.3 for AlK and CuL X rays, respectively, were determined after irradiation at 277 K using the technique of pulsed-field gel electrophoresis. Induction of DSBs is linearly dependent on dose. Oxygen enhancement ratios of 1.9 and 2.1 for cellular inactivation and DSB induction, respectively, were obtained with AIK X rays. These values are less than those for 60Co gamma radiation. The repair kinetics for rejoining of DSBs after a dose of 15 Gy is similar for both X-ray energies and 60Co gamma rays with a first half-life of 18-22 +/- 5 min. From these studies, it is suggested that induction of DSBs by low-LET radiations such as 60Co gamma rays reflects clustered damage produced predominantly by low-energy electron "track ends," which represent about 30% of the total dose. PMID- 9339948 TI - Effect of hydroxyl radical scavenging capacity on clustering of DNA damage. AB - We have shown previously that the thiol N-(2'-mercaptoethyl)-1,3-diaminopropane (WR-1065) can attenuate the formation of strand breaks associated with ionizing radiation. The mechanism of this protection is predominantly the reduction of DNA radical species which otherwise would attenuate the chemical repair of DNA radical species which are strand break precursors. We had observed that the presence of a hydroxyl radical scavenger during irradiation resulted in a decrease in the ability of WR-1065 to attenuate the formation of strand breaks. Since ionic compounds are known to affect the binding of the dicationic WR-1065 with the polyanion DNA, the effect of the scavenger was initially attributed to its polar nature having a similar effect on the interaction of WR-1065 with DNA, and not as a consequence of its ability to scavenge hydroxyl radicals. After examining additional scavengers, we now conclude that an increased hydroxyl radical scavenging capacity does attenuate the repair of strand break precursors to some extent. The probable explanation for this observation is that an increased scavenging capacity results in a greater degree of radical clustering on the DNA, and that these clusters of multiple radicals are repaired more slowly than are single radical species. PMID- 9339949 TI - Intra-arm and interarm chromosome intrachanges: tools for probing the geometry and dynamics of chromatin. AB - Many chromosome-type, exchange-type chromosomal aberrations produced by radiation are intrachanges, i.e. involve only one chromosome. It is assumed such intrachanges are formed by illegitimate reunion of two double-strand breaks (DSBs) on the chromosome. The yield of intra-arm intrachanges (acentric rings or paracentric inversions) relative to that of interarm intrachanges (centric rings or pericentric inversions) is larger than would occur if production and illegitimate reunion of DSBs were spatially random. The excess of intra-arm intrachanges is presumably due to proximity effects for illegitimate reunions, i.e. enhancement of the intrachange probability when two DSBs are formed close to one another. Radiation track structure may also play a role. Using a polymer description for "large-scale" chromatin geometry (>2 Mb), and using two alternate (rapid or slow motion) models for the way that DSBs move after they are produced, theoretical estimates are given for size distributions of intrachanges at low or high linear energy transfer (LET). The ratio of intra-arm to interarm intrachanges is derived from the size distribution and compared with data from the literature on centric rings, inversions, interstitial deletions and excess acentric fragments. Proximity effects enhance yields of intra-arm relative to interarm intrachanges at least severalfold and perhaps as much as 10-fold compared to expectations based on spatial randomness. We argue that further measurements of intra-arm and interarm intrachanges would be informative about large-scale chromatin structure and chromosome motion. Because inversions are more frequent than estimates of randomness would indicate, and are transmissible to daughter cells, their size distribution could also help characterize past exposure to high-LET radiation. PMID- 9339950 TI - The micronucleus assay of lymphocytes is a useful predictive assay of the radiosensitivity of normal tissue: a study of three inbred strains of mice. AB - Radiation sensitivity was measured by the micronucleus assay in T lymphocytes of the spleen and in fibroblasts derived from three inbred strains of mice, BALB/c, A/J and C3H/He. A linear correlation between the surviving fraction and the micronucleus frequency was obtained. There were clear differences in the radiosensitivity of T lymphocytes and of fibroblasts from the different strains of mice. T lymphocytes and fibroblasts from BALB/c mice were more radiosensitive than those from A/J and C3H/He mice. To explore the correlation between the radiosensitivity of T lymphocytes and of fibroblasts in vitro and the reaction of normal tissue to radiation, the survival ofjejunal crypt stem cells from the three mouse strains was determined using a microcolony assay. The jejunal crypt cells of BALB/c mice were more radiosensitive than those of A/J and C3H/He mice. Thus the order of radiosensitivity of T lymphocytes and fibroblasts in vitro corresponds to the sensitivity ofjejunal crypt cells in vivo. Our results suggest that determining the radiosensitivity of T lymphocytes and fibroblasts in vitro may be useful in predicting the magnitude of radiation-induced damage in the small intestine. PMID- 9339952 TI - Early damage to lung tissue after irradiation detected by the magnetic resonance T2 relaxation time. AB - We sought to determine whether nuclear magnetic resonance relaxation times of water in tissue would be useful to detect molecular damage in lung tissue within 2 weeks after irradiation. Tissue samples were obtained from the lungs of rats at various times between 1 and 14 days after exposure of a hemithorax to 20 Gy 60Co gamma irradiation. The spin-lattice relaxation time, T1, was measured by the inversion recovery method, and the spin-spin relaxation time, T2, was measured by both the Hahn spin-echo (Hahn T2) and the Carr-Purcell-Meiboom-Gill (CPMG T2) methods. The T2 of lung tissue could be divided into two components, T2 fast (T2f) and T2 slow (T2s), which reflected changes in the intracellular and extracellular water, respectively. The CPMG T2f increased significantly 3 days after irradiation (66.3 +/- 2.3 ms compared to 60.8 +/- 2.6 ms), and the CPMG T2s increased significantly 1 day after irradiation (155 +/- 11 ms compared to 138 +/ 7 ms), prior to the observation of abnormalities upon examination of the lung by light microscopy. The CPMG T2 values increased further up to 14 days after irradiation when significant increases were observed in values for T1, Hahn T2 and water content. Our results indicate that the molecular derangement in irradiated lung tissue was detected by the CPMG T2 measurement in the very early stage, and that MRI may be superior to conventional radiographs for detecting the early damage to lung tissue after irradiation. PMID- 9339951 TI - Analysis of the incidence of solid cancer among atomic bomb survivors using a two stage model of carcinogenesis. AB - A two-stage stochastic model for carcinogenesis was used to analyze the incidence of cancer of the lung, stomach and colon in the cohort of atomic bomb survivors. We fitted the model assuming that acute exposure to radiation results in the creation of initiated cells that are added to the pool of spontaneously initiated cells. In the cancers analyzed, with the exception of lung cancer in females, we found no evidence that radiation-induced initiation was dependent upon age at exposure. In contrast, we found that spontaneous initiation was dependent upon age at exposure in the cancers analyzed except stomach cancer among males. Because exposure to radiation in this cohort occurred at the same time for all members of the cohort, age at exposure is exactly correlated with birth cohort, and the dependence of spontaneous initiation on age at exposure is a reflection of the cohort effects seen in these cancers in Japan. Even without a dependence of radiation-induced initiation on age at exposure, the two-stage model can explain the temporal behavior of the excess relative risk with age at exposure and time since exposure. In particular, the model predicts that excess relative risk is highest among those exposed as children. Moreover, since radiation induced initiation is not higher among those exposed as children, the excess relative risk in this group is not due to an inherently higher sensitivity to radiation. Our biologically based approach provides another perspective on the temporal behavior of risk after acute exposure to ionizing radiation. PMID- 9339953 TI - Biological effects of inhaled 238PuO2 in beagles. AB - Beagle dogs exposed to 238PuO2 aerosols (136 dogs, 13-22 per group, mean initial lung depositions of 0.0, 0.13, 0.68, 3.1, 13, 52 and 210 kBq) were observed throughout life to determine tissues at risk and dose-effect relationships. The pulmonary retention of 238Pu was represented by the sum of two exponentially decreasing components of the initial lung deposition; about 84% cleared with a 174-day half-time; the half-time of the remainder was 908 days. The average percentages of final body burden found in lung, skeleton, liver and thoracic lymph nodes in the 30 longest-surviving dogs (mean survival 14 years) were 1, 46, 42 and 6%, respectively. Of 116 beagles exposed to plutonium, 34 (29%) developed bone tumors, 31 (27%) developed lung tumors, and 8 (7%) developed liver tumors. Although lungs accumulated a higher average radiation dose than skeleton, more deaths were due to bone tumors than to lung tumors. Deterministic effects included radiation pneumonitis, osteodystrophy, hepatic nodular hyperplasia, lymphopenia, neutropenia and sclerosing tracheobronchial lymphadenitis. Hypoadrenocorticism was also observed in a few dogs. Increased serum alanine aminotransferase, indicative of liver damage, was observed in groups with > or =3.1 kBq initial lung deposition. Estimates of cumulative tissue dose in a human exposed to airborne 238PuO2 for 50 years at a rate of one annual limit on intake each year were derived based on a comparison of the data on metabolism for humans and beagles. The 50-year dose estimates for humans are an order of magnitude lower than doses at which increased incidence of neoplasia was observed in these dogs, whereas the projected doses to humans from 50-year exposure at the annual limit of intake are of similar magnitude to those at which deterministic effects were seen in the beagles. PMID- 9339954 TI - The effect of the irradiation wavelength on the processes sensitized by protoporphyrin IX dimethyl ester. AB - The formation of triplet states of photosensitizers and singlet oxygen during reactions sensitized by protoporphyrin IX dimethyl ester (PPDME) and the products of its photo-oxidation in solution were studied by time-resolved spectroscopy. Irradiation with long-wavelength light (670 nm), which is absorbed by the products of photo-oxidation of PPDME, provides lower quantum yields of singlet oxygen than in sensitization with PPDME alone, which absorbs light with a wavelength of 630 nm. Spectroscopic measurements also confirmed the lower rate of sensitized photo-oxidation of bilirubin during irradiation with light with a wavelength of 670 nm. PMID- 9339955 TI - Predictions of mathematical models of tissue oxygenation and generation of singlet oxygen during photodynamic therapy. AB - Photodynamic therapy (PDT) is a relatively new protocol for cancer treatment which has recently been approved for limited clinical use. Traditionally, the success of treatment with PDT has been compared on the basis of total light delivery. Using the mathematical model of Henning et al. (Radiat. Res. 142, 221 226, 1995), we have determined that when oxygen is not depleted from the tissue, the concentration of singlet oxygen that is generated is directly proportional to the product of the light fluence rate (phi) and the concentration of the photosensitizer (Cs). Therefore, phiCs is an appropriate parameter for comparing the potential success of PDT protocols under these conditions. For a treatment of time t, the observed photodynamic effect resulting from singlet oxygen exposure should be directly related to phiCst. For high phiCs, the model predicts that oxygen depletion occurs within the tumor tissue. As a result, the photodynamic effect is no longer proportional to phiCst. We have expanded the model of Henning et al. to include the changes in oxygen concentration which occur within the capillary as blood flows through the tissue. Our new predictions with the mathematical model for optimal PDT treatment conditions are significantly different from those predicted by the previous models. Predictions of the model are given using parameters relevant for treatment of solid tumors with Photofrin. PMID- 9339956 TI - Induction of a particular deletion in mitochondrial DNA by X rays depends on the inherent radiosensitivity of the cells. AB - We examined whether X radiation induces a particular deletion in the mitochondrial DNA (mtDNA) of the cells of two human squamous cell carcinoma lines with different sensitivity to radiation and in a radiosensitive ataxia telangiectasia (AT) cell line. We used polymerase chain reaction (PCR) to quantify the accumulation of a particular 4977-bp deletion (delta mtDNA4977). PCR products of delta mtDNA4977 were detectable after exposure to 10 Gy in the radioresistant squamous cell carcinoma cells, 2 Gy in the radiosensitive squamous cell carcinoma cells and 1 Gy in the radiosensitive AT cells. These observations suggest that ionizing radiation induces the delta mtDNA4977 in human cells and that the radiation doses required to induce this deletion reflect the sensitivity of cells to radiation. PMID- 9339957 TI - Comments on the article "Studies of the mortality of atomic bomb survivors. Report 12, part I. Cancer: 1950-1990" by D. A. Pierce, Y. Shimizu. D. L. Preston, M. Vaeth and K. Mabuchi (Radiat. Res. 146, 1-27, 1996) PMID- 9339958 TI - The role of beta-blockers in left ventricular dysfunction and heart failure. AB - It was first reported by our group in 1975 that heart failure due to idiopathic dilated cardiomyopathy (IDC) could be improved by long term treatment with a beta blocker, starting at a low dose and continuing with a stepwise up-titration. Since then, many studies have been performed in patients with heart failure of various aetiologies and the beneficial effects of long term beta-blockade have been confirmed. About 3000 patients have been included in randomised studies in which beta-blockade, given for more than 2 months, mostly elicited significant improvements in functional class, exercise capacity, cardiac function, quality of life and/or morbidity. When started at a very low dose (one-tenth to one twentieth of the doses generally used in angina or hypertension), the treatment is well tolerated in most patients. In these studies, various types of beta blockers were used, including beta1-selective blockers and nonselective blockers with additional properties (vasodilator and antioxidative) such as metoprolol, bisoprolol, bucindolol and carvedilol. Several large studies have also reported benefits on mortality and morbidity. In the Metoprolol in Dilated Cardiomyopathy (MDC) trial, metoprolol treatment in patients with IDC resulted in a 34% reduction of the primary combined endpoint, total number of deaths and need for cardiac transplantation. In the Cardiac Insufficiency Bisoprolol Study (CIBIS), in patients with idiopathic as well as ischaemic cardiomyopathy, there was a nonsignificant 20% reduction in mortality. In the US carvedilol studies (n = 1094), also in patients with ischaemic and idiopathic cardiomyopathy, carvedilol reduced mortality by 65%, which was highly significant. A nonsignificant reduction in mortality was observed in the Australia-New Zealand (ANZ) Heart Failure Study with carvedilol. In all these studies there was a reduction in hospitalisations, with all drugs being generally well tolerated. It can thus be concluded that the beneficial effects of beta-blockers on cardiac function and morbidity have been documented in a large number of studies in selected groups of patients. The treatment has been accepted in some countries by the regulatory authorities. Larger, placebo-controlled studies are needed to convincingly demonstrate a reduction in total mortality as observed in the pooling of the 4 US carvedilol studies. Such studies are in progress for various beta-blockers, which may lead to acceptance of their routine clinical use in patients with congestive heart failure. PMID- 9339961 TI - Practical recommendations for the treatment of ascites and its complications. AB - Ascites is one of the earliest and most common complications of patients with cirrhosis, and is associated with complications such as dilutional hyponatraemia, renal dysfunction and spontaneous bacterial peritonitis. The treatment of ascites has been based on the combination of a low-sodium diet and the administration of diuretics. The reintroduction of paracentesis and the recent introduction of the transjugular intrahepatic portosystemic shunt (TIPS) are the most relevant innovations in the treatment of ascites during the past 2 decades. The development of ascites is closely related to renal disturbances of functional origin, including the hepatorenal syndrome. A new definition of hepatorenal syndrome has been proposed recently and 2 different types have been defined (type I or progressive, and type III or stable). Although no effective therapy exists for this syndrome, the use of therapeutic methods (TIPS, vasoconstrictor agents, dialysis) to temporarily improve renal function and act as a 'bridge' to liver transplantation, may be of most benefit. The use of potent and safe antibiotics has improved the resolution rate and survival of patients with spontaneous bacterial peritonitis. In addition, the use of oral antibiotics will simplify the management of this condition in the near future. Finally, prophylactic antibiotic regimens represent a major step forwards in the prevention of spontaneous bacterial peritonitis in subsets of cirrhotic patients with a great risk of developing this complication. PMID- 9339959 TI - Apoptosis: clinical relevance and pharmacological manipulation. AB - Apoptosis, often synonymously used with the term 'programmed cell death', is an active, genetically controlled process that removes unwanted or damaged cells. Suppression, overexpression or mutation of a number of genes which orchestrate the apoptotic process are associated with disease. The diseases in which apoptosis has been implicated can be grouped into 2 broad groups: those in which there is increased cell survival (i.e. associated with inhibition of apoptosis) and those in which there is excess cell death (where apoptosis is overactive). Diseases in which there is an excessive accumulation of cells include cancer, autoimmune disorders and viral infections. Deprivation of trophic factors is known to induce apoptosis in cells dependent on them for survival. This fact has been exploited in the use of antiandrogens or antiestrogens in the management of prostate or breast cancer. Haemopoietic growth factors like granulocyte macrophage colony stimulating factor (GM-CSF) or interleukin-3 prevent apoptosis in target cells and modulation of levels of these factors has been tried in the prevention of chemotherapy-induced myelosuppression. Until recently, it was thought that cytotoxic drugs killed target cells directly by interfering with some life-maintaining function. However, of late, it has been shown that exposure to several cytotoxic drugs with disparate mechanisms of action induces apoptosis in both malignant and normal cells. Physiological regulation of cell death is essential for the removal of potentially autoreactive lymphocytes during development and the removal of excess cells after the completion of an immune response. Recent work has clearly demonstrated that dysregulation of apoptosis may underlie the pathogenesis of autoimmune diseases by allowing abnormal autoreactive lymphocytes to survive. AIDS and neurodegenerative disorders like Alzheimer's or Parkinson's disease represent the most widely studied group of disorders where an excess of apoptosis has been implicated. Amyotrophic lateral sclerosis, retinitis pigmentosa, epilepsy and alcoholic brain damage are other neurological disorders in which apoptosis has been implicated. Apoptosis has been reported to occur in conditions characterised by ischaemia, e.g. myocardial infarction and stroke. The liver is a site where apoptosis occurs normally. This process has also been implicated in a number of liver disorders including obstructive jaundice. Hepatic damage due to toxins and drugs is also associated with apoptosis in hepatocytes. Apoptosis has also been identified as a key phenomenon in some diseases of the kidney, i.e. polycystic kidney, as well as in disorders of the pancreas like alcohol-induced pancreatitis and diabetes. PMID- 9339963 TI - Insulin lispro: a review of its pharmacological properties and therapeutic use in the management of diabetes mellitus. AB - Insulin lispro, a recombinant insulin analogue, is identical to human insulin except for the transposition of proline and lysine at positions 28 and 29 in the C-terminus of the B chain. The resultant reduced capacity for self-association in solution translates into more rapid absorption of insulin lispro than human regular insulin from subcutaneous sites. Maximum insulin concentrations are higher and are reached earlier with insulin lispro than with human regular insulin, and insulin concentrations return to baseline values more quickly with insulin lispro; consequently, insulin lispro has a more rapid onset and a shorter duration of glucose-lowering activity. These pharmacological properties provided the rationale for comparative clinical trials of subcutaneous insulin lispro (administered within 15 minutes before meals, preferably immediately before meals) and subcutaneous human regular insulin (administered 20 to 45 minutes before meals) in patients with type 1 diabetes (insulin-dependent diabetes mellitus) or type 2 diabetes (non-insulin-dependent diabetes mellitus) requiring premeal insulin therapy plus basal insulin therapy. Available clinical trials are well designed and results suggest that 1- and 2-hour postprandial blood glucose levels with insulin lispro are similar to or lower than those with human regular insulin; 1- and 2-hourpostprandial glucose excursions are similar to or less pronounced than those with human regular insulin. Glycated haemoglobin A values were generally similar with both agents. Continuous subcutaneous insulin infusion was associated with greater improvements in postprandial blood glucose levels and glycated haemoglobin A1 values with insulin lispro than with human regular insulin. Confirmatory data are required. The incidence of hypoglycaemia with insulin lispro was similar to or lower than that with human regular insulin. In particular insulin lispro appears to be associated with a lower incidence of night-time and severe hypoglycaemic episodes. Evidence also suggests that patients perceive their quality of life to be improved with insulin lispro compared with human regular insulin, and that satisfaction with treatment is greater with the insulin analogue. Thus, in patients with type 1 or 2 diabetes requiring premeal insulin therapy, insulin lispro appears to provide greater postprandial glycaemic control than human regular insulin without increasing the risk of hypoglycaemia. Furthermore, the reduced injection-meal interval with this agent offers greater convenience for the patient than regular human insulin. If longer term clinical experience supports these promising results it is likely that insulin lispro will offer important advantages over human regular insulin. PMID- 9339962 TI - Current guidelines on stress ulcer prophylaxis. AB - Acute uppergastrointestinal bleeding in intensive care unit (ICU) patients may occur due to peptic ulcer disease, adverse drug effects, gastric tube lesions, acute renal failure, liver failure or stress-induced gastric mucosal lesions. Gastric acid hypersecretion can be observed in patients with head trauma or neurosurgical procedures. Gastric mucosal ischaemia due to hypotension and shock is the most important risk factor for stress ulcer bleeding. Preventive strategies aim to reduce gastric acidity (histamine H2 receptor antagonists, antacids), strengthen mucosal defensive mechanisms (sucralfate, antacids, pirenzepine) and normalise gastric mucosal microcirculation (sucralfate, pirenzepine). However, the most important prophylactic measure is an optimised resuscitation and ICU regime aiming to improve oxygenation and microcirculation. All drugs approved for stress ulcer prophylaxis in Europe (H2 antagonists, antacids, pirenzepine, sucralfate) have been shown to be effective in prospective controlled randomised trials. However, due to insufficient clinical data, prostaglandins and omeprazole cannot be recommended for this use. Stress ulcer prophylaxis is indicated only in patients at risk, and not in every ICU patient. The selection of drugs today depends not only on efficacy but also on possible adverse effects and on costs. In this regard, the most cost-effective drug is sucralfate. The clinical relevance of nosocomial pneumonia due to gastric bacterial overgrowth has decreased during the past decade due to several changes in the management of critically ill patients. PMID- 9339965 TI - Fenoldopam: a review of its pharmacodynamic and pharmacokinetic properties and intravenous clinical potential in the management of hypertensive urgencies and emergencies. AB - Fenoldopam is a dopamine agonist that causes peripheral vasodilation via stimulation of dopamine 1 (D1) receptors. The efficacy of an intravenous infusion of fenoldopam in decreasing blood pressure in patients with a hypertensive urgency, including patients who developed hypertension after coronary artery bypass graft surgery, and in a small number of patients with hypertensive emergency, is similar to that of sodium nitroprusside. However, unlike sodium nitroprusside, fenoldopam also increases renal blood flow and causes diuresis and natriuresis. There is no evidence of rebound hypertension after stopping the infusion. As the tolerability profile of fenoldopam is generally similar to that of sodium nitroprusside, fenoldopam appears to be an effective alternative to sodium nitroprusside in the immediate treatment of patients who develop severe hypertension and in whom oral treatment is not practical. Fenoldopam may be particularly useful in patients who develop hypertension after coronary artery bypass graft surgery, but further studies are required to confirm its role in hypertensive emergency. PMID- 9339960 TI - Prevention of transplant rejection: current treatment guidelines and future developments. AB - In the past 2 decades, progressive improvements in the results of organ transplantation as a therapeutic strategy for patients with end-stage organ disease have been achieved due to greater insight into the immunobiology of graft rejection and better measures for surgical and medical management. It is now known that T cells play a central role in the specific immune response of acute allograft rejection. Strategies to prevent T cell activation or effector function are thus all potentially useful for immunosuppression. Standard immunosuppressive therapy in renal transplantation consists of baseline therapy to prevent rejection and short courses of high-dose corticosteroids or monoclonal or polyclonal antibodies as treatment of ongoing rejection episodes. Triple-drug therapy with the combination of cyclosporin, corticosteroids and azathioprine is now the most frequently used immunosuppressive drug regimen in cadaveric kidney recipients. The continuing search for more selective and specific agents has become, in the past decade, one of the priorities for transplant medicine. Some of these compounds are now entering routine clinical practice: among them are tacrolimus (which has a mechanism of action similar to that of cyclosporin), mycophenolate mofetil and mizoribine (which selectively inhibit the enzyme inosine monophosphate dehydrogenase, the rate-limiting enzyme for de novo purine synthesis during cell division), and sirolimus (rapamycin) [which acts on and inhibits kinase homologues required for cell-cycle progression in response to growth factors, like interleukin-2 (IL-2)]. Other new pharmacological strategies and innovative approaches to organ transplantation are also under development. Application of this technology will offer enormous potential not only for the investigation of mechanisms and mediators of graft rejection but also for therapeutic intervention. PMID- 9339964 TI - Micronised fenofibrate: a review of its pharmacodynamic properties and clinical efficacy in the management of dyslipidaemia. AB - Micronised fenofibrate is a new formulation of the fibric acid derivative fenofibrate. It is indicated for the treatment of patients with type IIa, IIb, III or IV dyslipidaemia who have failed to respond to dietary control or other nonpharmacological interventions. Micronised fenofibrate has improved absorption characteristics compared with the standard preparation, allowing a lower daily dosage and once-daily administration. The lipid-modifying profile of micronised fenofibrate is characterised by a decrease in low density lipoprotein (LDL) and total cholesterol levels, a marked reduction in elevated plasma triglyceride levels and an increase in high density lipoprotein (HDL) cholesterol levels. Consistent with the standard formulation, which is administered as 300mg daily in divided doses, the micronised preparation has demonstrated efficacy in the treatment of type IIa, IIb and IV primary dyslipidaemias but at a lower daily dosage of 200mg once daily. Because of its significant triglyceride-lowering effect, micronised fenofibrate appears to be of greatest benefit in patients with hypertriglyceridaemia (with or without hypercholesterolaemia), including patients with type 2 (non-insulin-dependent) diabetes mellitus and dyslipidaemia. In the comparisons available, micronised fenofibrate 200mg once daily was of similar efficacy to or less effective than the HMG-CoA reductase inhibitors simvastatin 20mg daily and pravastatin 20mg daily at reducing LDL and total cholesterol levels. However micronised fenofibrate produced greater improvements in triglyceride and, generally, HDL cholesterol levels than both simvastatin and pravastatin. Data on the long term tolerability of micronised fenofibrate are limited. However, data from a large short term (3-month) study have indicated that gastrointestinal disorders are the most frequent adverse events associated with therapy. Elevations in serum transaminase and creatine phosphokinase levels have been reported rarely with micronised fenofibrate. In conclusion, available data suggest that the more convenient lower once-daily dosage of micronisedfeno fibrate retains the beneficial lipid-modifying effects of the standard formulation. Further studies are required to determine whether the lipid changes achieved with micronised fenofibrate result in a reduction in cardiovascular morbidity and mortality. PMID- 9339967 TI - The validity of self-reported condom use among adolescents. AB - BACKGROUND: Research and public health interventions designed to reduce the risk of sexually transmitted diseases (STDs) often are based on self-reported condom use. Yet, validation of self-reported condom use, in particular with adolescents, has rarely been described in the literature. METHODS: Baseline data were obtained from 540 adolescents, 13-21 years of age, enrolled in a 1-year longitudinal study of health beliefs, sexual behaviors, and STD acquisition. Of the 445 participants reporting to be sexually active, 404 (90.8%) agreed to a complete physical examination, including a genital examination, with STD screening after completing the self-administered written questionnaire. Participants' written self-report of condom use was compared to histories obtained by clinicians and laboratory diagnosis of acute STDs to assess validity of written self-report. RESULTS: Complete data were available for 321 females and 77 males of whom 52 females and 5 males had laboratory evidence of 63 infections. Although three individuals who had STDs reported to be consistent users of condoms, a significant association (P < 0.05) was found between those who reported more frequent condom use with the last two partners and the absence of STDs. CONCLUSION: In this group of adolescents, self-report of condom use with the last two partners was associated with the absence of an acute STD. This finding suggests that self-reported condom use is a valid indicator of risk for STDs, with implication for those working with adolescents clinically and in research contexts. PMID- 9339966 TI - Chlamydial and gonococcal cervicitis in HIV-seropositive and HIV-seronegative pregnant women in Bangkok: prevalence, risk factors, and relation to perinatal HIV transmission. AB - OBJECTIVES: To determine the prevalence and risk factors associated with cervicitis caused by Chlamydia trachomatis and Neisseria gonorrhoeae in human immunodeficiency virus (HIV) type 1-seropositive and HIV-seronegative pregnant women in Bangkok, and the relation to perinatal HIV transmission. METHODS: As part of a multicenter perinatal HIV transmission study in an antenatal population with 2% HIV seroprevalence, endocervical swabs obtained at mid-pregnancy from a consecutive sample of 222 HIV-seropositive and 219 HIV-seronegative pregnant women at two large hospitals in Bangkok were tested for the presence of C. trachomatis and N. gonorrhoeae by DNA hybridization probe (Gen-Probe). Clinical risk factors and DNA probe results were analyzed in relation to the women's and newborns' HIV infection status. RESULTS: The prevalence of C. trachomatis was 16.2% in HIV-seropositive pregnant women and 9.1% in HIV-seronegative pregnant women (P = 0.03). The prevalence of N. gonorrhoeae was 2.7% in HIV-seropositive pregnant women and 1.4% in HIV-seronegative pregnant women (P = 0.5). The overall population prevalence estimate was 9.2% for C. trachomatis and 1.4% for N. gonorrhoeae. Women with gonococcal infection were more likely to be positive for C. trachomatis (RR(MH) = 5.2, P < 0.01). Young age (<21 years) and primigravid status were associated with C. trachomatis infection among HIV-seropositive women; history of multiple sex partners (>1) were associated with C. trachomatis infection among HIV-seronegative women. For HIV-seropositive women, primigravida status also was associated with C. trachomatis infection. The perinatal HIV transmission rates were similar for those with and without C. trachomatis (24.1% and 23.2%, P = 0.9) and among those with and without N. gonorrhoeae (20% and 23.5%, P = 1.0). CONCLUSIONS: Among pregnant women in Bangkok, C. trachomatis infection was considerably more common than N. gonorrhoeae infection and was associated with HIV infection, young age and first pregnancy (HIV-seropositive women), and multiple partners (HIV-seronegative women). Our data do not suggest an association between perinatal HIV transmission and maternal C. trachomatis or N. gonorrhoeae infection identified and treated during pregnancy. The high prevalence of C. trachomatis found using a test not readily available in Thailand emphasizes the need for improved, inexpensive ways to screen for and diagnose these sexually transmitted infections in developing countries. PMID- 9339968 TI - Partner notification for syphilis: a randomized, controlled trial of three approaches. AB - OBJECTIVE: To determine the cost and effectiveness of three approaches to partner notification for infectious syphilis. STUDY DESIGN: People with syphilis were randomly assigned to: (1) notification of partners by patients themselves within 2 days or disease intervention specialists would notify them; (2) immediate notification by intervention specialist; or (3) immediate notification by intervention specialists, who had the option of drawing blood in the field. Costs of intervention specialists' time, travel, and overhead were measured. Intention to-treat analysis measured outcomes per randomized index patient. RESULTS: From December, 1990 through March, 1993, 1,966 index patients with syphilis (primary 9%; secondary 18%; and early latent 73%) were randomized in Broward County (Ft. Lauderdale), Florida (1,191); Tampa, Florida (569); and Paterson, New Jersey (206). Index patients reported 11,272 potentially exposed partners and sufficient information to initiate investigations for 2,761. Of these, 2,236 were located, 367 had newly identified infections, and 870 others received preventive treatment. The three partner notification approaches had similar success locating partners (1.1-1.2 per index patient) and treating partners (0.61-0.67 per index). The cost was $317 to $362 per partner treated; the optimal strategy differed by study site. CONCLUSIONS: Partner notification identified many infected and potentially infected people. The cost and effectiveness of the three types of provider notification were similar. Alternative approaches are needed to reach infected partners who could not be notified. PMID- 9339969 TI - Contact tracing's price is not its value. PMID- 9339970 TI - Zenilman's anomaly reconsidered: fallible reports, ceteris paribus, and other hypotheses. AB - BACKGROUND AND OBJECTIVES: In the January-February, 1995 issue of Sexually Transmitted Diseases, Zenilman and colleagues reported a null association between incident sexually transmitted diseases (STDs) and self-reported condom use. That anomalous finding generated a flurry of letters to the editor, some of which were quite heated. This article reconsiders the Zenilman team's results. STUDY DESIGN: New statistical analyses were conducted to test two hypotheses that sought to account for the null association: (1) deviation from study protocol, and (2) differential risks of acquiring an incident STD among segments of the study population that varied by reported level of condom use. RESULTS: No support was found for hypotheses concerning deviation from study protocol and differential risk of acquiring an incident STD by level of condom use. Indeed, for respondents who reported multiple sexual partners, the analyses found increased rates of infection among those who reported more consistent condom use. CONCLUSIONS: Two of the most promising hypotheses for explaining Zenilman's anomalous findings are unsupported by reanalysis of the available empirical evidence. It is still possible that respondents who reported that they used condoms consistently differed from self-reported nonusers or inconsistent users in some way that altered their risk of acquiring an STD and thus obscured the protective effects of properly used condoms. Nonetheless, as Zenilman and others suggest, fallibility in self-reports of condom use remains the primary suspect as the cause of these anomalous results. Such fallibility may be particularly pronounced when self-reported behavioral data are collected in contexts that include strong educational campaigns or other norm-setting interventions. PMID- 9339971 TI - Risk factors for sexually transmitted disease in Harare: a case-control study. AB - OBJECTIVE: To obtain information on risk factors and health-seeking behavior of men with sexually transmitted diseases (STDs) attending primary care clinics. STUDY DESIGN: Unmatched case-control. METHODS: Cases consisted of 256 consecutive men with genital ulcer disease (GUD) and 256 with other STDs. Control subjects (N = 256) were recruited from every third man with non-STD-related complaints. All subjects were at least 15 years of age. A structured questionnaire was administered. RESULTS: Genital ulcer disease cases reported more frequent sexual intercourse with a commercial sex worker (odds ratio [OR] = 17.4; 95% confidence intervals [CI] = 7.8-40.0) and a recent new sexual contact (OR = 6.7; CI = 3.3 14.1). Similarly, STD cases reported more frequent sexual contact with a commercial sex worker (OR = 3.4; CI = 2.0-5.6) and a recent new sexual contact (OR = 7.9; CI = 3.9-16.3). Reported condom use was less than 30% with all partner types. Of all STD cases, 80% sought treatment at the primary care clinics, with 35% delaying more than 7 days before seeking treatment. CONCLUSIONS: Culturally appropriate behavioral educational programs are advocated to reduce the risk of transmission and the period for seeking treatment for all STDs. PMID- 9339972 TI - Incidence and determinants of hepatitis C virus infection among individuals at risk of sexually transmitted diseases attending a human immunodeficiency virus type 1 testing program. AB - BACKGROUND: The role of sexual transmission of hepatitis C virus (HCV) infection is still not completely understood, partly because of the lack of longitudinal studies among cohorts of HCV-negative individuals who engage in at-risk sexual behavior. GOALS: To evaluate the incidence of HCV infection in a population at risk for human immunodeficiency virus type 1 (HIV-1) infection and other sexually transmitted diseases (STD) and to identify factors associated with HCV seroconversion. STUDY DESIGN: A retrospective longitudinal study was carried out on a cohort of consecutive attendees of a voluntary HIV-1 testing and counseling program in a large STD center in Rome. All individuals undergoing at least two consecutive tests for HCV antibodies were enrolled. Clinical data and information on individual behavior were collected for all study participants. RESULTS: Between June, 1992 and December, 1994, a total of 709 individuals (12 intravenous drug users [IDU], 244 homosexuals, and 453 heterosexual non-IDUs), initially negative for HCV antibody, were tested more than once. Among these individuals, 15 HCV seroconversions occurred. The average follow-up time was 1.25 person/years (p/y) for an incidence rate of 1.69 per 100 p/y. The incidence rates by exposure category were 39.30 per 100 p/y among IDUs, 1.37 per 100 p/y among homosexual men, and 0.97 per 100 p/y among heterosexual non-IDUs. Excluding IDUs, of the 697 STD clinic attendees engaging in at-risk sexual behavior, HIV-1-positive status tended to be associated with HCV seroconversion (relative hazard = 5.48; 95% confidence interval = 0.85-35.40). The HCV crude incidence rates among HIV-1 infected patients at enrollment was 11.5%, 4.2%, and 2.4% in those with severe, moderate, and mild levels of immunosuppression, respectively (chi-square for trend = 2.38, P = 0.1). CONCLUSIONS: In this cohort, HCV infection was confirmed to be strongly associated with intravenous drug use. Nonetheless, the occurrence of two thirds of the total HCV seroconversions in non-IDU individuals engaging in at-risk behavior suggests a role of sexual practices in the transmission of the infection. Among non-IDU individuals, the risk for development of HCV infection tended to increase in those who were HIV-1 infected. PMID- 9339973 TI - Prevalence of Chlamydia trachomatis infection in a low-risk population in Hungary. AB - BACKGROUND AND OBJECTIVE: Chlamydia trachomatis is the leading cause of nongonococcal urethritis and cervicitis in women. Because of the recent increases in the numbers of new cases and severe consequences, there is an urgent demand for the introduction of sensitive and specific rapid diagnostic methods. GOAL: A multicenter examination involving seven centers was sponsored by the Hungarian Ministry of Health and Welfare in order to provide a survey of Chlamydia trachomatis in the gravid population. 6,161 women were tested between 1994 to 1995. STUDY DESIGN: The seven centers were selected with regard to different aspects, from developed and less developed areas in the capital, two large provincial towns, and various other provincial regions reflecting either an industrial or an agricultural background. The nucleic acid hybridization method (PACE 2 Gen-Probe, San Diego, CA) was introduced in this low-risk population for the examination of Chlamydia trachomatis. In one center, a further two methods, antigen detection by ELISA (SYVA, CA) and cultivation on the McCoy cell line (staining with SYVA FITC-labeled antichlamydia monoclonal antibody), were applied. RESULTS: International surveys and experience indicate that the proportion of the population threatened by Chlamydia trachomatis is above 10%. The overall average incidence of Chlamydia trachomatis cases in this low-risk gravid population was 5.74%. The data from the different centers ranged between 1.6% and 9.7%. The chlamydia-infected Hungarian gravid population is below the critical 10%, but there is one Hungarian county where the value is close to 10%. CONCLUSIONS: In this provincial, industrial area, the number of unmarried and divorced gravida in a low economic situation is disproportionately high. For this disadvantaged population, permanent Chlamydia trachomatis screening was suggested. In the other centers, screening of pregnant women for Chlamydia trachomatis and the treatment of positive cases and their partners were suggested for pathological gravida with preterm labor and preterm rupture of the membranes. PMID- 9339974 TI - Trichomoniasis in a postmenopausal woman cured after discontinuation of estrogen replacement therapy. AB - BACKGROUND: Epidemiologic and experimental data suggest that estrogen has a salutary effect on Trichomonas vaginalis. CASE: A metronidazole-allergic postmenopausal woman was cured of vaginal trichomoniasis in association with discontinuation of estrogen replacement therapy. CONCLUSION: Hormonal manipulation should be studied for the management of women with trichomoniasis who are allergic to metronidazole or who are infected with metronidazole resistant strains of Trichomonas vaginalis. PMID- 9339975 TI - Multiple condom use in commercial sex in Lamphun Province, Thailand: a community generated STD/HIV prevention strategy. AB - BACKGROUND AND OBJECTIVE: In an investigation of condom breakage in commercial sex, we found a high proportion of multiple condoms use. This study sought to ascertain the characteristics of brothel-based commercial sex workers (CSWs) and their clients; to identify the decision makers (clients and/or CSWs) active in choosing multiple condom use; and to determine whether there is an implicit hierarchy of condom use negotiation. GOALS: To identify factors associated with multiple condom use in commercial sex and to provide an understanding of how this innovation developed in this setting. STUDY DESIGN: Sixty-seven brothel-based CSWs in Lamphun Province who participated in a study of condom breakage participated in a case-control study of multiple versus single condom use, which determined CSW and client characteristics for evidence of multiple condom use. Interviews and focus groups were used to determine decision making for condom use and the contexts for multiple use. Association between characteristics of CSWs/clients and multiple condom use was analyzed using X2 for trend. RESULTS: No official program encouraged multiple condom use; this appeared to be a community devised strategy to increase protection from human immunodeficiency virus (HIV) and other sexually transmitted diseases (STDs). CSWs working in daytime brothels had significantly greater multiple condom use than those working in nighttime brothels; mean percentages + SD of multiple simultaneous use were 57.5% + 28.6 and 35.5% + 22.4, respectively (p < 0.001). Day CSWs were older, had more pregnancies, reported higher frequency of STD symptoms and history of pelvic inflammatory disease, and had more clients per day but had a lower number of sex acts per client than night CSWs. Among factors associated with multiple condoms, only age was significant. The decision to use single or multiple condoms for a sex act was primarily (78.2% in single and 79.3% in multiple) made by the CSW herself. The main reason given for multiple condom use was protection from HIV/STD. CONCLUSIONS: There is high compliance between CSWs in Lamphun province and the Ministry of Public Health-sponsored 100% condom use campaign, and CSWs are attempting to further reduce their risks of HIV/STD exposure by using multiple condoms for sex with their clients. PMID- 9339976 TI - Would you like to use one condom or two? PMID- 9339977 TI - Sexual network data help assess putative STD reporting bias. PMID- 9339978 TI - Mitochondrial DNA deletions associated with aging and presbyacusis. AB - BACKGROUND: The membrane hypothesis of aging proposes an association between reactive oxygen metabolites and aging processes. Reactive oxygen metabolites are a normal by-product of oxidative phosphorylation and are also formed under conditions of ischemia, hypoperfusion, and as a result of environmental contaminants. Among the many detrimental activities of reactive oxygen metabolites, also known as free oxygen radicals, is direct damage to mitochondrial DNA. Progressive accumulation of mitochondrial DNA damage renders cells unable to conduct oxidative phosphorylation reactions effectively, thereby leading to a bioenergetically deficient cell. Over time, mitochondrial DNA damage accumulates and leads to cellular dysfunction with subsequent organ failure, aging, and ultimately, death. This sequence forms the basis of the membrane hypothesis of aging. OBJECTIVE: To determine if the membrane hypothesis of aging may be involved in the development of presbyacusis. DESIGN: Fischer rats from 4 age groups were tested for auditory sensitivity using the auditory brainstem response. Brain, stria vascularis, and auditory nerve tissues were harvested and mitochondrial DNA was amplified to identify the highly conserved cytochrome b and ND1-16S ribosomal RNA segment of the NADH genes, as well as a 4834-base pair (bp) deletion associated with aging. SUBJECTS: Fischer rats (n=28) from 4 age groups were used: young (2-4 months [n=9]), mid-young (9-11 months [n=5]), mid-old (18 20 months [n=5]), and old (30-34 months [n=9]). RESULTS: The results demonstrate a progressive reduction in auditory sensitivity with age. The mitochondrial DNA studies identify a significant increase in the presence of the 4834-bp deletion in the aged subjects compared with the young. CONCLUSIONS: These findings raise the possibility that the 4834-bp deletion may be associated with presbyacusis, as well as with aging. PMID- 9339979 TI - Quality of life for children with otitis media. AB - OBJECTIVE: To evaluate changes in health-related quality of life for children with otitis media. DESIGN: Cohort study using a 6-item quality-of-life survey (OM 6) representing the domains of physical suffering, hearing loss, speech impairment, emotional distress, activity limitations, and caregiver concerns. SETTING: Hospital-based pediatric otolaryngology practice in a metropolitan area. PATIENTS: One hundred eighty-six children aged 6 months to 12 years (median age, 3.4 years) with chronic otitis media with effusion or recurrent acute otitis media. INTERVENTION: The OM-6 was completed at entry by the child's caregiver and again at least 4 weeks after routine clinical care. Otoscopic findings, static admittance, tympanometric width, audiometric thresholds, and ear-related global quality of life (10-point visual scale) were recorded concurrently. MAIN OUTCOME MEASURES: Test-retest reliability, construct validity, and responsiveness to longitudinal change of the OM-6 survey score (mean value of the 6 items). RESULTS: Excellent test-retest reliability was obtained for the survey score (R=0.87) and individual survey items (R> or =0.71). The median survey score was 2.8 (95% confidence interval, 2.7-3.0) of a maximum 7.0, with higher values indicating poorer quality of life. Construct validity was shown by significant correlations between the survey score and global ear-related quality of life (R= 0.64), between physical suffering and physician visits in the past month (R=0.47), and between caregiver concerns and antibiotics consumed in the past month (R=0.26). The mean change in survey scores after tympanostomy tubes was 1.7, with a standardized response mean of 1.7 (95% confidence interval, 1.4-2.0), indicating large responsiveness to change. The change score was reliable (R=0.82) and correlated well with the degree of reported clinical change (R=0.66). CONCLUSIONS: The OM-6 is a valid, reliable, and responsive measure of quality of life for children with otitis media. The brevity and ease of administration make the OM-6 ideal for use in outcomes studies, clinical trials, and routine clinical care. PMID- 9339981 TI - Prognostic factors in the treatment of lymphatic malformations. AB - OBJECTIVE: To find factors that may influence the treatment outcomes of lymphatic malformations of the head and neck in children. DESIGN: Charts of patients treated surgically for lymphatic malformations of the head and neck between 1988 and 1996 at our tertiary care children's hospital were reviewed retrospectively. Outcomes were correlated with age at presentation, associated symptoms, anatomical site (s) of involvement, extent of disease, length of time between first symptoms and surgery, completeness of removal, and histologic pattern. PATIENTS: Of 85 children treated, 74 underwent primary surgical excision at our hospital. Follow-up ranged from 6 months to 8 years, with a mean of 3 years. RESULTS: The overall recurrence rate, judged by functional or cosmetic deformity, was 22%. Two neonates died of the disease. Factors associated with a better prognosis were a single anatomical site of involvement; location in the neck, even if involving 2 sites; and the impression of completeness of resection at the time of surgery. Findings associated with a higher recurrence rate included younger age (especially neonates) and the presence of associated symptoms (ie, infection, dyspnea, dysphagia, and hemorrhage). The histologic pattern and the length of time from diagnosis to treatment were not significantly associated with the prognosis. CONCLUSIONS: We recommend aggressive, timely surgical excision for lymphatic malformations of the head and neck. The timing of surgery should be based on the child's functional and cosmetic deformity at the time of presentation and on the likelihood of complete excision, weighed against the morbidity associated with surgical excision. PMID- 9339980 TI - Chronic otitis media treated topically with ciprofloxacin or tobramycin. AB - OBJECTIVE: To evaluate the efficacy of ciprofloxacin compared with tobramycin and placebo ear drops in the treatment of chronic suppurative otitis media without cholesteatoma. DESIGN: Sixty ears (in 51 patients) were randomly divided into 3 treatment groups: ciprofloxacin hydrochloride, tobramycin, and placebo interventions. SETTING: The otolaryngology department of a university teaching hospital. INTERVENTION: All ears were treated topically for 3 weeks. MAIN OUTCOME MEASURES: Each patient received a small, numbered bottle and was instructed to instill 5 drops 3 times daily for 3 weeks. The final clinical and bacteriologic assessment was made after 3 weeks. RESULTS: The organism most commonly isolated from the ear discharge was Pseudomonas aeruginosa. Its sensitivity to ciprofloxacin and tobramycin was 94.2% and 70.6%, respectively. The clinical response was 78.9%, 72.2%, and 41.2% in the ciprofloxacin, tobramycin, and placebo groups, respectively. The bacteriologic response rate was 66.7% for the ciprofloxacin and tobramycin groups and 20% for the placebo group. Treatment with ciprofloxacin ear drops seemed to be as effective as treatment with tobramycin. CONCLUSION: While the lack of ototoxicity of ciprofloxacin was not tested in our study, this treatment may be considered as a potential topical therapy for cases of chronic suppurative otitis media. PMID- 9339982 TI - A simple method for closure of tracheocutaneous fistula in children. AB - OBJECTIVE: To review the results of a simple technique of closure of persistent tracheocutaneous fistula (TCF) in children. DESIGN: Retrospective case series. SETTING: Tertiary pediatric otolaryngology referral center. PATIENTS: Children (age, < 18 years) who underwent repair of TCF from July 1, 1991, to August 31, 1996. INTERVENTIONS: Surgical closure of persistent TCF using multilayered closure of de-epithelialized local tissue. Tracheal dissection was not performed. A thermal hemostatic scalpel was used in some cases to assist in de epithelialization and provide hemostasis without electrocautery near the airway. MAIN OUTCOME MEASURES: Success of closure and number and types of complications. RESULTS: Nine procedures were performed in 8 children. Seven (88%) of 8 primary procedures were successful, but early recurrent TCF developed in 1 patient. Revision surgery using an identical surgical technique, but maintaining endotracheal intubation for 48 hours, was successful in this patient. No complications occurred. CONCLUSIONS: This procedure is a simple, reliable method for closure of TCF in children. PMID- 9339983 TI - Histopathologic changes after pericardial patch tracheoplasty. AB - Pericardial patch tracheoplasty has been used for surgical correction of long segment congenital tracheal stenosis caused by complete tracheal rings in infants. The case histories of 2 infants with descriptions of the histopathologic changes in their respective tracheas are presented. Complete reepithelialization of the graft site with ciliated pseudostratified columnar epithelium was found, suggesting the likelihood of normal mucociliary flow in the trachea. The pericardial patches were replaced by mature scar tissue in the graft site, establishing a functional tracheal lumen. Wound healing in the trachea is reviewed. Obstruction by exuberant granulation tissue is an ongoing concern. Pericardium continues to be an important option as graft material for tracheal reconstruction in infants with long-segment congenital tracheal stenosis. PMID- 9339984 TI - Cricotracheal anastomosis for assisted ventilation-induced stenosis. AB - OBJECTIVE: To review the long-term results and our experience with cricotracheal anastomosis via a cervical approach for assisted ventilation-induced stenosis. DESIGN: A case series of 41 patients consecutively treated with cricotracheal anastomosis. SETTING: A tertiary care center and university teaching hospital. PATIENTS: Group 1 consisted of 22 patients with stenosis reaching the lower border of the cricoid cartilage that did not require resection of the cricoid cartilage. Group 2 consisted of 19 patients in whom correction of the stenosis required cricoid resection. MAIN OUTCOME MEASURES: Statistical analysis of airway patency was based on the Kaplan-Meier actuarial life table method. Incidence for the various postoperative complications was presented. Univariate analysis was performed to analyze the relationships between various factors, airway patency, and the incidence for the various complications encountered. RESULTS: The Kaplan Meier 5-year airway patency estimate was 100% in group 1 patients and 82.5% in group 2 patients. In group 2 patients, complementary treatment with dilatations in 2 patients resulted in an overall 94.8% airway patency rate. In the last patient, the airway patency was not reestablished after cricotracheal anastomosis, and a Montgomery T tube was inserted. Postoperative complications included unilateral inferior laryngeal nerve paralysis (3 patients), cervical neck abscess (2 patients), pneumothorax (1 patient), and major subcutaneous emphysema (1 patient). None of the following variables was statistically related to the airway patency or to the various complications encountered: sex, age, cause for stenosis, delay from initial injury, prior treatment, presence of a tracheotomy, number of tracheal rings resected, type of sutures used, and type of anastomosis performed. CONCLUSIONS: The data reported reemphasized that cricotracheal anastomosis with or without cricoid resection is a safe and reliable procedure for assisted ventilation-induced upper tracheal stenosis reaching and/or involving the subglottis and/or cricoid cartilage. PMID- 9339985 TI - Clinical outcome of continuous facial nerve monitoring during primary parotidectomy. AB - OBJECTIVES: To assess whether continuous facial nerve monitoring during parotidectomy is associated with a lower incidence of facial nerve paresis or paralysis compared with parotidectomy without monitoring and to assess the cost of such monitoring. DESIGN: A retrospective analysis of outcomes for patients who underwent parotidectomy with or without continuous facial nerve monitoring. SETTING: University medical center. PATIENTS: Fifty-six patients undergoing parotidectomy in whom continuous electromyographic monitoring was used and 61 patients in whom it was not used. MAIN OUTCOME MEASURES: (1) The incidence of early and persistent facial nerve paresis or paralysis and (2) the cost associated with facial nerve monitoring. RESULTS: Early, unintentional facial weakness was significantly lower in the group monitored by electromyograpy (43.6%) than in the unmonitored group (62.3%) (P=.04). In the subgroup of patients without comorbid conditions or surgeries, early weakness in the monitored group (33.3%) remained statistically lower than the rate of early weakness in the unmonitored group (57.5%) (P=.03). There was no statistical difference in the final facial nerve function or incidence of permanent nerve injury between the groups or subgroups. After multivariate analysis, nonmonitored status (odds ratio [OR], 3.22), advancing age (OR, 1.47 per 10 years), and longer operative times (OR, 1.3 per hour) were the only significant independent predictive variables significantly associated with early postoperative facial weakness. The incremental cost of facial nerve monitoring was $379. CONCLUSIONS: The results suggest that continuous electromyographic monitoring of facial muscle during primary parotidectomy reduces the incidence of short-term postoperative facial paresis. Advantages and disadvantages of this technique need to be considered together with the additional costs in deciding whether routine use of continuous monitoring is a useful, cost-effective adjunct to parotid surgery. PMID- 9339986 TI - Facial nerve function after parotidectomy. AB - OBJECTIVES: To analyze the incidence of facial nerve dysfunction following parotidectomy and to correlate this with the extent of parotid gland resection, the pathological diagnosis, and the clinical setting. DESIGN: A review of prospectively collected data from a dedicated computerized head and neck database. SETTING: Tertiary care center. PATIENTS: Between 1987 and 1995, 248 patients underwent 259 parotidectomies performed by the same surgeon (C.J.O'B.). Indications were clinical tumor (n=213) or sialadenitis (n=46). There were 235 previously untreated patients and 13 who had undergone a prior operation on that side. Facial nerve function was normal in 242 patients and abnormal before surgery in 6. Cancers accounted for 88 parotidectomies and benign disease accounted for 171. Of 213 clinical tumors, 41 (19%) were situated deep to the plane of the facial nerve. RESULTS: The facial nerve was intentionally sacrificed in 28 of 259 operations (18 total and 10 partial sacrifice). In 230 parotidectomies in which facial nerve function was normal before surgery and the nerve was preserved, the incidence of initial postoperative facial weakness was 29%. Based on the diagnosis and extent of surgery, rates of facial weakness were 16.5% and 13%, respectively, for benign and malignant tumors located in the superficial lobe and treated with limited superficial parotidectomy; 30% and 34% for sialadenitis treated with complete superficial parotidectomy and near-total parotidectomy, respectively; 31% and 100%, respectively, for benign and malignant lobe tumors treated with near-total parotidectomy; 83% for parotidectomy associated with a neck dissection; and 33% for patients who had previous parotid surgery. Permanent weakness occurred in 13 (5.6%) of 230 patients, but 10 of these 13 had simultaneous neck dissection and facial nerve dysfunction involved only the marginal mandibular branch. Recovery of normal facial movements occurred within 6 months in 46 (68%) of 67 of those with initial weakness. CONCLUSIONS: The likelihood of temporary facial weakness correlated with the extent of surgery and was especially influenced by tumor location deep to the plane of the facial nerve, previous parotid surgery, a diagnosis of sialadenitis, and the addition of neck dissection to the parotidectomy. Permanent weakness mainly affected the marginal mandibular branch when neck dissection was included. PMID- 9339987 TI - Effects of functional endoscopic sinus surgery on maxillary sinus mucosa. AB - OBJECTIVE: To compare the preoperative and postoperative changes on ciliary surface of maxillary sinus mucosa in patients treated with functional endoscopic sinus surgery. DESIGN: The maxillary mucosa of both the superolateral wall and the ostium were sampled during the operation and 6 to 12 months (mean duration, 7.6 months) after the operation. Ciliary surface was determined using scanning electron microscopy in combination with an image analyzer and was expressed in terms of ciliary area, which is the percentage of mucosal surface occupied by cilia. SETTING: The samples were taken at a hospital-based clinic. An electron microscopic study was performed at Mie University School of Medicine, Mie, Japan. PATIENTS: Sixteen patients (20 maxillary sinuses) undergoing functional endoscopic sinus surgery for treatment of chronic sinusitis. RESULTS: The mean (+/-SD) ciliary area before the surgery was 60.7%+/-28.8% and 39.9%+/-21.5% in the superolateral wall of the maxillary sinus and the ostium of the maxillary sinus, respectively. The ciliary area of the superolateral wall was significantly higher than that of the ostium (P<.001). The mean (+/-SD) postoperative ciliary area value was 74.3%+/-22.6% in the superolateral wall and 51.3%+/-16.1% in the ostium. These postoperative values were significantly higher than the preoperative values (P<.001). CONCLUSIONS: This study showed that the maxillary sinus mucosa in chronic sinusitis is capable of regeneration, and the damaged ciliated epithelium could return toward normal with the improvement of ventilation and drainage of the maxillary sinus following functional endoscopic sinus surgery. PMID- 9339989 TI - The role of dental prostheses in alveolar ridge squamous carcinomas. AB - BACKGROUND: Alveolar ridge squamous carcinomas develop in patients outside the usual constellation of risk factors. OBJECTIVE: To determine whether the use of dentures was a risk factor specific to patients with alveolar ridge carcinoma. DESIGN: Case-control method with a unique control group-a concurrent cohort of patients with head and neck cancer with primaries in the oropharynx, hypopharynx, and larynx. SETTING: Tertiary care hospital-based clinic. PATIENTS: Forty-one patients with squamous carcinomas centered on the maxillary or mandibular alveolar ridges. The control group was 175 concurrently seen patients with squamous carcinomas of the laryngopharynx for whom dental status was known. MAIN OUTCOME MEASURES: Age at diagnosis, sex, tobacco use, alcohol use, and denture use. RESULTS: Patients with alveolar ridge were more likely to be female, older, nonsmokers, and nondrinkers. The crude odds ratio of denture use in patients with alveolar ridge cancer was 2.28 (P=.03). Eliminating other confounding factors with logistic regression, the adjusted odds ratio dropped to 1.30 (P=.59). Among patients with alveolar ridge, smoking status correlated with age and gender: current smokers were on average 64.4 years old and 9 of 16 were men. Nonsmokers' average age was 79.1 years and 1 of 11 was a man. CONCLUSIONS: In this study, denture use was not an independent risk factor for alveolar ridge carcinomas. Among patients with little to no tobacco or alcohol exposure, the alveolar ridge carcinomas tended to occur in the elderly and in women. PMID- 9339988 TI - RANTES is more prevalent in bacterial than in nonbacterial maxillary sinusitis: and P-selectin is preferentially up-regulated in diseased mucosae. AB - OBJECTIVE: To investigate the relationship of the clinical appearance, histological characteristics, bacterial culturing, and messenger RNA (mRNA) expression of RANTES, interleukin 6, and interleukin 12, as well as the occurrence of endothelial adhesion molecules, in inflammatory diseased maxillary sinus mucosa in critically ill patients. DESIGN: Prospective case series. SETTING: General intensive care unit and neurosurgical intensive care unit of a tertiary care hospital. SUBJECTS: Seven critically ill patients, nasotracheally intubated or tracheotomized, who received ventilator treatment for more than 7 days and treatment with antibiotics. INTERVENTIONS: Bilateral biopsy specimens of antral mucosa were obtained at sinoscopy. Reverse transcriptase-polymerase chain reaction was used to detect the cytokine mRNAs in situ on paraformaldehyde-fixed tissue, and intercellular adhesion molecule 1, vascular cell adhesion molecule 1, E-selectin, and P-selectin were analyzed by immunochemistry on frozen sections. Sampling of secretion and tissue from the antra was performed for bacterial culturing. RESULTS: Macroscopic and histological appearance varied and showed moderate to pronounced inflammation in 6 antra. All 4 bacterially infected antra showed mRNA RANTES (P=.005). No correlation was found for interleukin 6 and interleukin 12. Up-regulation of P-selectin in all cases and sparse expression of vascular cell adhesion molecule 1 indicate that the inflammation is chronic but nonallergic in type. CONCLUSION: We find an indication that RANTES is more prevalent in bacterial sinusitis. PMID- 9339991 TI - Head and neck cancer-specific quality of life: instrument validation. AB - BACKGROUND: The disfigurement and dysfunction associated with head and neck cancer affect emotional well-being and some of the most basic functions of life. Most cancer-specific quality-of-life assessments give a single composite score for head and neck cancer-related quality of life. OBJECTIVE: To develop and evaluate an improved multidimensional instrument to assess head and neck cancer related functional status and well-being. METHODS: The item selection process included literature review, interviews with health care workers, and patient surveys. A survey with 37 disease-specific questions and the SF-12 survey were administered to 253 patients in 3 large medical centers. Factor analysis was performed to identify disease-specific domains. Domain scores were calculated as the standardized score of the component items. These domains were assessed for construct validity based on clinical hypotheses and test-retest reliability. RESULTS: Four relevant domains were identified: Eating (6 items), Communication (4 items), Pain (4 items), and Emotion (6 items). Each had an internal consistency (Cronbach alpha value) of greater than 0.80. Construct validity was demonstrated by moderate correlations with the SF-12 Physical and Mental component scores (r=0.43-0.60). Test-retest reliability for each domain demonstrated strong reliability between the 2 time points. Correlations were strong for each individual question, ranging from 0.53 to 0.93. Construct validity testing demonstrated that the direction of differences for each domain were as hypothesized. CONCLUSION: The Head and Neck Quality of Life questionnaire is a promising multidimensional tool with which to assess head and neck cancer specific quality of life. PMID- 9339990 TI - Clinical-severity staging system for oropharyngeal cancer: five-year survival rates. AB - OBJECTIVE: To improve the classification and survival estimates for patients with oropharyngeal cancer by combining cancer symptom severity and comorbidity with the current TNM cancer staging system. DESIGN: Retrospective medical record review using explicit coding criteria. SETTING: University medical center. PATIENTS AND METHODS: Two hundred ninety-six patients receiving initial treatment from January 1, 1980, to December 31, 1989. Multivariate analysis identified patient factors that had a significant impact on 5-year survival. These patient factors, symptom severity and comorbidity, were combined with cancer stage to create a composite clinical-severity staging system. MAIN OUTCOME MEASURE: Five year survival. RESULTS: The overall 5-year survival rate was 38% (111/ 296). Survival by TNM cancer stage was 67% (18/27) for stage I, 46% (24/52) for stage II, 31% (26/85) for stage III, and 32% (43/132) for stage IV (chi2=10.84; P=.001). When patients were grouped according to the clinical-severity staging system, survival rates were 70% (16 of 23) for stage A, 47% (71 of 152) for stage B, 27% (18 of 67) for stage C, and 11% (6 of 54) for stage D (chi2=34.49; P=.001). CONCLUSIONS: Survival estimates can be improved by adding carefully studied and suitably defined patient variables to the TNM cancer stage. The current TNM cancer staging system for oropharyngeal cancer is based solely on the morphologic description of the tumor and disregards the clinical condition of the patient. Cancer symptom severity and comorbidity have a significant impact on survival. Continued exclusion of patient factors leads to imprecision in prognostic estimates and hinders interpretation of clinical studies. PMID- 9339992 TI - Dominant hereditary conductive hearing loss due to an ossified stapedius tendon. AB - Familial conductive deafness is rare. This report confirms the existence of a lineage with congenital conductive hearing loss in 3 generations. The results of otologic evaluations, including pure-tone audiometry, tympanometry, acoustic reflex test, and liquid test, in 14 patients in this family were consistent with the findings of ossicular fixation in 10 patients. Tympanotomies performed in both ears of 4 patients indicated stapes ankylosis caused by ossification of the stapedius tendon. The patients gained normal hearing levels using a simple surgical procedure. PMID- 9339993 TI - Pathologic quiz case 1. Primary systemic amyloidosis in a patient with multiple myeloma. PMID- 9339994 TI - Pathologic quiz case 2. Castleman disease (giant lymph node hyperplasia). PMID- 9339995 TI - The liposhaver in facial plastic surgery. PMID- 9339996 TI - Proceedings of the 3rd WONOEP Symposium. Tromso, Norway, June 1993. PMID- 9339997 TI - Report of The International Working Group on Harmonization of Dementia Drug Guidelines. PMID- 9339998 TI - Mechanical Circulatory Support. Proceedings of the 5th International Symposium. Bad Oeynhausen, 7-9 September 1995. PMID- 9339999 TI - Proceedings of the 1996 International Symposium on Subsurface Microbiology (ISSM 96). Davos, Switzerland, 15-21 September 1996. PMID- 9340000 TI - Biotransformation and biodegradation of N-substituted aromatics in methanogenic granular sludge. AB - N-Substituted aromatic compounds are environmental contaminants associated with the production and use of dyes, explosives, pesticides and pharmaceuticals. In this article, we examine the potential of anaerobic granular sludge from anaerobic treatment systems towards the detoxification, transformation, and mineralization of nitroaromatic and azo compounds. Nitroaromatics and azo dyes with strong electron withdrawing are highly inhibitory to acetoclastic methanogenic bacteria. However, nitro and azo substituted aromatics are readily reductively detoxified in methanogenic consortia to their respective aromatic amines, which are several orders of magnitude less toxic. This reductive detoxification has allowed the successful operation of anaerobic reactors for the treatment of highly toxic aromatic compounds. In the course of the experiments it was discovered that some aromatic amines were mineralized. These results indicate that some N-substituted aromatic compounds can be completely mineralized and serve as a carbon and energy source for anaerobic bacteria. PMID- 9340001 TI - Degradation of ethylenethiourea (ETU) in oxic and anoxic sandy aquifers. AB - Ethylenethiourea is an important degradation product of ethylenebisdithiocarbamate fungicides, which are widely used in different kinds of crops. The ethylenebisdithiocarbamate group includes maneb, zineb and mancozeb. The ethylenebisdithiocarbamates are not highly toxic and degrade rapidly in the presence of moisture and oxygen, forming different compounds. One of these is the polar ethylenethiourea, which is relatively stable. Thus, this compound appears to be a potential contaminant for groundwater. Batch experiments were carried out under biotic as well as abiotic conditions to study the degradation dependence of concentration, temperature and organic matter. The decomposition of ethylenethiourea under abiotic conditions was found to be less than 5% of the degradation under biotic conditions. Further, ethylenethiourea showed to be stable over a period of 150 days at 20 degrees C in tap water as well as in batch with soil sterilized with NaN3. The degradation of ethylenethiourea depends on the concentration in the water implying first order reaction kinetics. The microbial degradation of ethylenethiourea is highly temperature dependent with aerobic Q10 between 2.9 and 4.2, and an anaerobic between 2.1 and 2.5. A minor increase in degradation rates was observed by application of nitrate and manure to the batches. The experiments show extremely complete degradation of ethylenethiourea in the presence of microbial nitrate reduction with pyrite which occurs in deeper parts of the aquifers. PMID- 9340002 TI - Microbiology in nuclear waste disposal: interfaces and reaction fronts. AB - It is now generally acknowledged that microbial populations will be present within nuclear waste repositories and that the consequences of such activity on repository performance must be assessed. Various modelling approaches--based either on mass balance/thermodynamics or on kinetics--have been developed to provide scoping estimates of the possible development of these populations. Past work has focused on particular areas of the repository which can be considered relatively homogeneous and hence can be represented by some kind of 'box' or 'mixing tank'. In reality, however, waste repositories include a range of engineering materials (steel, concrete, etc.) which are emplaced at depth in a rock formation. Strong chemical gradients--of the type which may be exploited by lithoautotrophic microbial populations--are likely to be found at the contacts between different materials and at the interface between the engineered structures and the host rock. Over the long timescales considered, solute transport processes will cause the locations of strong chemical gradients to move, forming reaction fronts. The high-pH plume resulting from the leaching of cement/concrete in some repository types is a particularly important example of such a reaction front. Redox fronts, which may occur in different areas of all kinds of repositories, also play an important role and would be locations where microbial activity is likely to be significant. In this paper, the key microbial processes expected at (or around) interfaces and fronts will be discussed, with particular emphasis on the development of quantitative models. The applicability of the models used wil be tested by considering similar fronts which can be found in natural systems. PMID- 9340003 TI - DRINK: a biogeochemical source term model for low level radioactive waste disposal sites. AB - Interactions between element chemistry and the ambient geochemistry play a significant role in the control of radionuclide migration in the geosphere. These same interactions influence radionuclide release from near surface, low level radioactive waste, disposal sites once physical containment has degraded. In situations where LLW contains significant amounts of metal and organic materials such as cellulose, microbial degradation in conjunction with corrosion can significantly perturb the ambient geochemistry. These processes typically produce a transition from oxidising to reducing conditions and can influence radionuclide migration through changes in both the dominant radionuclide species and mineral phases. The DRINK (DRIgg Near field Kinetic) code is a biogeochemical transport code designed to simulate the long term evolution of the UK low level radioactive waste disposal site at Drigg. Drigg is the UK's principal solid low level radioactive waste disposal site and has been receiving waste since 1959. The interaction between microbial activity, the ambient geochemistry and radionuclide chemistry is central to the DRINK approach with the development of the ambient pH, redox potential and bulk geochemistry being directly influenced by microbial activity. This paper describes the microbial aspects of the code, site data underpinning the microbial model, the microbiology/chemistry interface and provides an example of the code in action. PMID- 9340004 TI - Symposium on sensorimotor circuits controlling behavioral functions in birds. In honour of Prof. Dr. Jacob L. Duddeldam. PMID- 9340005 TI - Papers from the annual congress of the Swiss Society of Pediatric Surgeons associated with the Swiss Society of Pediatrics and Swiss Society of Pediatric Anesthesia. Biel, Switzerland, June 20-22, 1996. PMID- 9340006 TI - ESEEM study of the phyllosemiquinone radical A1.- in 14N- and 15N-labeled photosystem I. AB - The phyllosemiquinone radical of the photosystem I reaction center has been studied by electron spin echo envelope modulation (ESEEM) spectroscopy. A comparative analysis of ESEEM data of the semiquinone in 14N- and 15N-labeled PSI and numerical simulations demonstrate the existence of two protein nitrogen nuclei coupled to the semiquinone. One of the 14N couplings is characterized by a quadrupolar coupling constant e2qQ/4h of 0.77 MHz, an asymmetry parameter eta of 0.18, and a hyperfine coupling tensor with an almost pure isotropic hyperfine coupling, i.e. (Axx, Ayy, Azz) = (1.3, 1.3, 1.5 MHz). The second nitrogen coupling is characterized by a quadrupolar coupling constant e2qQ/4h of 0.45 MHz, an asymmetry parameter eta of 0.85, and a weak hyperfine coupling tensor with a dominant anisotropic part, i.e. (Axx, Ayy, Azz) = (-0.2, -0.2, 1.5 MHz). On the basis of a comparison of the 14N-ESEEM data with 14N-NQR and 14N-ESEEM data from the literature, the first coupled nitrogen is assigned to the indole nitrogen of a tryptophan residue. The coupling of the second nitrogen is much weaker and therefore more difficult to assign. However, the simulated spectrum best describes an amino nitrogen of a histidine, although the amide group of an asparagine or glutamine cannot be ruled out. The possible origins of teh nitrogen hyperfine coupling are discussed in terms of the amino acid residues thought to be close to the semiquinone in PSI. PMID- 9340007 TI - Cardiac high molecular weight calmodulin binding protein contains calpastatin activity. AB - A high molecular weight calmodulin binding protein (HMWCaMBP) was previously identified and purified from bovine heart cytosolic fraction [Sharma, R.K. (1990) J. Biol. Chem. 265, 1152-1157]. In this study, we report the biological function of this protein. HMWCaMBP was subjected to peptide mapping and three peptides were sequenced. Two of the three peptide sequences were shown to be highly homologous to the calpain inhibitor, calpastatin. However, the third peptide did not show homology to any known proteins. The Western blot analysis of HMWCaMBP and purified calpastatin from bovine cardiac muscle showed immunoreactivity with polyclonal antibody raised against HMWCaMBP. Furthermore, HMWCaMBP inhibited calpain II and calpain I activities in a dose dependent fashion. Our data based on sequence homology, amino acid analysis, antibody reactivity and calpain inhibition suggests that HMWCaMBP is homologous to calpastatin and may be a CaM binding form of calpastatin. PMID- 9340008 TI - Pulsed EPR structure analysis of photosystem I single crystals: localization of the phylloquinone acceptor. AB - A novel application of electron paramagnetic resonance (EPR) is reported to gain three dimensional structural information on cofactors in proteins. The method is applied here to determine the unknown position of the electron acceptor QK, a phylloquinone (vitamin K1), in the electron transfer chain in photosystem I of oxygenic photosynthesis. The unusual electron spin echo (out-of-phase echo) observed for the light induced radical pair P700.+QK.- in PS I allows the measurement of the dipolar coupling between the two radical pair spins which yields directly the distance between these two radicals. Full advantage of the information in the out-of-phase echo modulation can be taken if measurements using single crystals are performed. With such samples, the orientation of the principal axis of the dipolar interaction, i.e., the axis connecting P700.+QK.-, can be determined with respect to the crystal axes system. An angle of theta = (27 +/- 5)degrees between the dipolar coupling axis and the crystallographic c axis has been derived from the modulation of the out-of-phase echo. Furthermore, the projection of the dipolar axis into the crystallographic a,b-plane, is found to be parallel to the a-axis. The results allow for the determination of two possible locations of QK within the electron transfer chain of photosystem I. These two positions are related to each other by the pseudo C2 symmetry of the chlorophyll cofactors. PMID- 9340009 TI - A thymocyte factor SATB1 suppresses transcription of stably integrated matrix attachment region-linked reporter genes. AB - SATB1 specifically recognizes and binds to specialized genomic regions with an ATC sequence context with high base-unpairing propensity. Such base-unpairing regions (BURs) are typically identified within nuclear scaffold- or matrix attachment regions (S/MARs). SATB1 is a homeodomain protein and is predominantly expressed in thymocytes. We obtained BHK cell lines expressing low levels of SATB1 by stable transfection and investigated its effect on stably integrated MAR linked SV40 enhancer/promoter-driven luciferase reporter genes. For this study, both naturally occurring and synthetic MARs, as well as an AT-rich non-MAR control, were tested. Previous studies demonstrated that MAR sequences augment transcription of the linked reporter luciferase gene. Here, we show that SATB1 dramatically reduces the high levels of MAR-linked luciferase gene transcription. Transcription was virtually abolished for a reporter gene surrounded by two MARs at the 5' and 3' ends of the gene, which otherwise confer the highest level of transcriptional augmentation. On the other hand, SATB1 did not affect expression of an AT-rich non-MAR-linked luciferase gene or of endogenous housekeeping genes. This study shows that SATB1 acts as a strong transcriptional suppressor on a reporter gene linked to MARs when it is stably integrated into chromatin. PMID- 9340010 TI - Ca2+-signaling cycle of a membrane-docking C2 domain. AB - The C2 domain is a Ca2+-dependent, membrane-targeting motif originally discovered in protein kinase C and recently identified in numerous eukaryotic signal transducing proteins, including cytosolic phospholipase A2 (cPLA2) of the vertebrate inflammation pathway. Intracellular Ca2+ signals recruit the C2 domain of cPLA2 to cellular membranes where the enzymatic domain hydrolyzes specific lipids to release arachidonic acid, thereby initiating the inflammatory response. Equilibrium binding and stopped-flow kinetic experiments reveal that the C2 domain of human cPLA2 binds two Ca2+ ions with positive cooperativity, yielding a conformational change and membrane docking. When Ca2+ is removed, the two Ca2+ ions dissociate rapidly and virtually simultaneously from the isolated domain in solution. In contrast, the Ca2+-binding sites become occluded in the membrane bound complex such that Ca2+ binding and dissociation are slowed. Dissociation of the two Ca2+ ions from the membrane-bound domain is an ordered sequential process, and release of the domain from the membrane is simultaneous with dissociation of the second ion. Thus, the Ca2+-signaling cycle of the C2 domain passes through an active, membrane-bound state possessing two occluded Ca2+ ions, one of which is essential for maintenance of the protein-membrane complex. PMID- 9340011 TI - [Vertebral body metastases: characteristic MRI findings in epidural infiltration]. AB - PURPOSE: In case of lumbar vertebral metastasis associated with anterior epidural carcinomatous infiltration, we have observed that infiltrations tend to respect the midline. This study led to the systematic recognition of these phenomena in vertebral metastases. MATERIALS AND METHODS: 11 patients with 17 vertebral metastases and adjacent anterior epidural infiltration were reviewed retrospectively. All cases were studied by MRI. The routinely used imaging technique included spin echo (SE) T1 and T2 weighted sequences in the sagittal plane native and T1-SE without and with Gd-DTPA in the axial planes. The radiological findings of these phenomena and the anatomy were studied. RESULTS: We observed these phenomena to be uni- or bilateral in 88.3% of all cases with intraspinal anterior epidural carcinomatous infiltration, especially in that part of the vertebral body where the basal vertebral venous plexus was located. CONCLUSION: We conclude that vertebral metastases respect the midline. We interpret this fact as being due the anatomy of the vertebral body and especially its stabilization by the posterior longitudinal ligament. These findings may be helpful in the differential diagnosis of vertebral body metastases with epidural infiltration in contrast to intraspinal processes which proceed with the destruction of the vertebral body. PMID- 9340012 TI - [Dependence of axis deviation in the esophagram on computerized tomography determined wall thickness and its value in assessing depth of invasion of esophageal carcinoma]. AB - PURPOSE: In this study we compared the abnormalities of the esophageal axis seen in the esophagogram with the thickness of the esophageal wall measured by computed tomography. We have investigated, how exactly both methods assess the local tumor invasion according to the TNM criteria and whether there is a relation between the esophageal axis and the wall thickness. METHODS: In a retrospective study we examined the esophagograms of 65 tumor patients. Computed tomography examinations were available in 40 cases. Using a graphical method the wall thickness was transferred to the esophagograms under consideration of the different scales of the images. RESULTS: There is no correlation between the different types of distortion of the esophageal axis and the wall thickness in computed tomography. However, it can be demonstrated that the distortion results from specific fixation effects with the surrounding tissue. CONCLUSIONS: Both radiological methods cannot determinate the tumor invasion correctly. PMID- 9340013 TI - [Severe obliteration of the urethral lumen after wall stent implantation. An unusual radiographic finding]. AB - The treatment of urethral stricture is still a challenge for urologists. Irrespective of the treatment employed, urethral stricture recurs in about 30% of all cases. In recent years, the wall stent, originally conceived for vascular surgery, has proved to be effective for the treatment of bulbar urethral strictures. The results are good, morbidity and complications occur only occasionally. In this paper, we described the case of a young patient who suffered from complete occlusion of the prosthesis 8 months after its implantation. The low age of the patient and the X-ray features of this case are unusual. The obstruction was successfully resolved by endoscopic resection. Follow-up after 14 months revealed a mild, short stenosis of the proximal tip. PMID- 9340014 TI - [Are nonionic contrast media identical in their tolerability? Results of a double blind randomized multicenter study with iomeprol and iopromide]. AB - Since a larger number of nonionic contrast media is available for the radiologist, the question arises as to whether they differ in their clinical tolerability. A double-blind, randomized, two-group comparison of phase IV with lomeprol and lopromide was carried out at 6 hospitals involving a total of 1,200 patients with the indication for computed tomography. The contrast media doses and the flow in computed tomography of the skull, thorax, and abdomen were, depending on the centre, between 50 and 200 ml and 0.5 and 3.0 ml/s, respectively. The biostatistical evaluation of adverse events which were probably contrast medium-related produced a highly significant difference between the two contrast media in favor of lomeprol (p = 0.0005). The difference in the reactions of heat, nausea, and vomiting is of clinical relevance as such adverse events may negatively affect the examination procedure and the opacification in spiral computed tomography. PMID- 9340015 TI - [Contrast media-associated nephropathy--pathogenesis and prevention]. AB - Contrast media-associated nephrotoxicity continues to be a relevant cause of acute renal failure, especially in patients with pre-existing renal insufficiency. Alterations in renal hemodynamics and direct tubular toxicity by contrast media are the primary factors believed to be responsible for contrast media-associated nephrotoxicity. We review recent insights into the pathogenesis of this complication and summarize prophylactic strategies focussing on hydration, vasoactive pharmacological agents, and prophylactic hemodialysis'. PMID- 9340016 TI - [Castleman lymphoma: isolated abdominal involvement with an 8-year course in a child]. AB - This article describes a child with isolated, abdominally localized Castleman's disease and 8-years follow-up. Six years after surgical resection of an adrenal tumour, a multifocal disease with liver, spleen and lymph node involvement developed, which has improved for now 2 years after steroid medication. PMID- 9340017 TI - [Pulmonary artery stenosis in aggressive mediastinal fibrosis; diagnosis and 3D imaging with helical CT examination]. AB - An aggressive mediastinal fibrosis was found in a 42-year-old female, suffering from dysphagia, stabbing pain in the chest, and an unclear weight loss. In this case, the rare combination of esophageal involvement, bronchial narrowing, and pulmonary artery obstruction could easily be demonstrated with a barium study and a helical CT examination including three-dimensional reconstructions. PMID- 9340018 TI - [Monostotic Paget's disease as differential diagnosis in osteolysis of the tibia]. AB - Monostotic Paget's disease has a well known appearance in radiography. Only in exceptions is a further diagnostic procedure necessary. The value of MRI and scintigraphy is demonstrated on an osteolytic lesion of the tibia. PMID- 9340021 TI - [Comparison of film-screen combination in a contrast detail diagram and with interactive image analysis. 1: Contrast detail diagram]. AB - The following three film-screen combinations were compared: a) a combination of anticrossover film and UV-light emitting screens, b) a combination of blue-light emitting screens and film, and c) a conventional green fluorescing screen film combination. Radiographs of a specially designed plexiglass phantom (0.2 x 0.2 x 0.12 m3) were obtained that contained bar patterns of lead and plaster (calcium sulfate) to test high and intermediate contrast resolution and bar patterns of air to test low contrast resolution, respectively. An aluminum step wedge was integrated to evaluate dose-density curves of the radiographs. The dose values for the various step thicknesses were measured as percentage of the dose value in air for 60, 81, and 117 kV. Exposure conditions were the following: 12 pulse generator, 0.6 mm focus size, 4.7 mm aluminum prefilter, a grid with 40 lines/cm (12:1), and a focus-detector distance of 1.15 m. The thresholds of visible bars of the various pattern materials were assessed by seven radiologists, one technician, and the authors. The resulting contrast detail diagram could not prove any significant differences between the three tested screen film combinations. The pairwise comparison, however, found 8 of the 18 paired differences to be statistically significant between the conventional and the two new screen-film combinations. The authors concluded that subjective visual assessment of the threshold in a contrast detail study alone is of only limited value to grade image quality if no well-defined criteria are used (BIR report 20 [1989] 137-139). The statistical approach of paired differences of the estimated means appeared to be more appropriate. PMID- 9340020 TI - [Studies of image quality and dose in digital subtraction angiography]. AB - METHODS: The dose needed for high image quality in l.a. DSA, was determined by investigation of the necessary relationship between dose and image quality. This method is based on measurements of the signal-to-noise ratio and of the contrast detail detectability. It has been tested on two different systems (Philips integris V3000, Philips Diagnost 97). RESULTS: Our measurements prove that an image intensifier entrance dose of 1.1 microGy (image intensifier diameter 38 cm) is sufficient for high image quality in DSA. An increased dose value does not lead to an improvement in image information. For details larger than the pixel size the 512 matrix produces a higher signal-to-noise ratio with a higher contrast-detail detectability, whereas higher spatial resolution results by the use of a 1024 matrix. In rotational angiography, high image quality can be achieved at an image intensifier entrance dose of 1.1 microGy. In contrast to sequences without rotation, image quality is decreased through movement unsharpness. As the maximum exposure time 25 ms should be selected. The use of a 1024 matrix does not lead to a better spatial resolution in rotational angiography. PMID- 9340019 TI - [Soft tissue emphysema as manifestation of perforating diverticulitis]. AB - Pronounced soft-tissue emphysema expressing disseminated fecal abscess formation in perforated colonic diverticulitis is described. Cervical, mediastinal, and thoracic dissemination and involvement of the scrotal space and the thigh reveal the original retroperitoneal localization as well as the unusual but potential dissemination. The diagnostic methods are described. PMID- 9340022 TI - [Virtual endoscopy with post-processing helical CT data sets]. AB - PURPOSE: The purpose of this work was to test a newly developed, post-processing software for virtual CT endoscopic methods. Virtual endoscopic images were generated from helical CT data sets in the region of the shoulder joint (n = 2), the tracheobronchial system (n = 3), the nasal sinuses (n = 2), the colon (n = 2), and the common carotid artery n = 1). Software developed specifically for virtual endoscopy ("Navigator") was used which, after a previous threshold value selection, makes the reconstruction of internal body surfaces possible by an automatic segmentation process. We have evaluated the usage of the software, the reconstruction time for individual images and sequences of images as well as the quality of the reconstruction. All pathological findings of the virtual endoscopy were confirmed by surgery. RESULTS: The post-processing program is easy to use and provides virtual endoscopic images within 50 seconds. Depending of the extent of the data set, virtual tracheobronchoscopy as a cine loop sequence required about 15 minutes. Through use of the threshold value-dependent surface reconstruction the demands on the computer configuration are limited; however, this also created quality problems in image calculation as a consequence of the accompanying loss of data. CONCLUSIONS: The Navigator software enables the calculation of virtual endoscopic models with only moderate demands on the hardware. PMID- 9340023 TI - [Image reconstruction of computerized tomography pictures using functional algebra]. AB - A detailed presentation of the process for calculating computed tomograms from the measured data by means of functional algebra is given and an attempt is made to demonstrate the relationships to those inexperienced in mathematics. Suggestions are also made to the manufacturers for improving tomography software although the authors cannot exclude the possibility that some of the recommendations may have already been realized. An interpolation in Fourier space to right-angled coordinates was not employed so that additional computer time and errors resulting from the interpolation are avoided. The savings in calculation time can only be estimated but should amount to about 25%. The error-correction calculation is merely a suggestion since it depends considerably on the apparatus used. Functional algebra is introduced here because it is not so well known but does provide appreciable simplifications in comparison to an explicit presentation. Didactic reasons as well as the possibility for reducing calculation time provided the foundation for this work. PMID- 9340024 TI - [Open questions on the topic of teleradiology from the current viewpoint]. AB - A review on the theoretical challenges and potential risks of teleradiology is given. Based on the authors two years clinical experience and the current literature unsolved problems are discussed, which are to a smaller extent the technical implementation but much more the still missing regulation of various concomitant circumstances. PMID- 9340025 TI - [Endothelin]. PMID- 9340026 TI - [Measuring core body temperature--a methodological challenge?]. PMID- 9340027 TI - [The significance of endothelin for physiologic and pathophysiologic processes of the lung]. AB - The endothelins (ETs) are a family of three vasoactive peptides (ET-1, ET-2, ET 3) that were first described in 1988. ETs have a wide range of action including vasoconstriction, vasodilatation, bronchoconstriction, and mitogenesis. Two types of ET receptors, classified as ETA and ETB receptors, have been identified in gene technology. Endothelins are produced by endothelial cells, smooth muscle cells, and bronchial epithelial cells. Their vasoactive effects contribute not only to homoeostasis but ETs seem also to be involved in several pulmonary diseases. Elevated ET plasma levels have been found in patients suffering from asthma, pulmonary fibrosis, pulmonary hypertension, and acute lung injury. This review gives a short summary of the actual facts in endothelin research, focussing on the effects of ET-1 in pulmonary circulation. PMID- 9340028 TI - [Temperature measurement in the ear canal: comparison of an infrared thermometer with conventional temperature probes and evaluation of clinical factors on infrared measurement]. AB - PURPOSE: We compared the readings of the GENIUS 3000A FirstTemp infrared thermometer with a conventional temperature probe (MON-A-THERM tympanic) placed in contact with the tympanic membrane. We also systematically evaluated user dependent factors influencing the infrared readings. METHODS: In 100 postoperative patients we investigated the repeatability of the infrared measurements and the agreement with tympanic thermocouple probes in the contralateral ear regarding the degree of auditory canal contamination. In 20 volunteers we evaluated the influence of three factors: interval between consecutive measurements, dwell time in the auditory canal before reading the temperature, and positioning of the grip. Finally, we compared the infrared readings in the same ear of 20 different users who were not familiar with this new method. RESULTS: Comparison with the tympanic contact probes revealed a mean difference of -0.67 degree C (+/-0.65 degree C 2 SD). The infrared thermometer significantly underestimated the temperature of the thermocouple probes. Repeatability was +/-0.3 degree C. The presence of cerumen in the auditory canal had no influence on the infrared readings. Shortening of the interval between two consecutive readings (30 and 60 s.) led to increasing differences between the two measurements with the second reading decreasing. After positioning the infrared thermometer in the auditory canal 5 seconds before taking temperatures, the recorded temperatures were significantly lower compared to the immediate temperature recordings. Rotation of the device out of the telephone handle position led to an increasing lack of agreement between infrared thermometry and thermistor probes. In 20 inexperienced operators agreement with the thermocouple probe was -0.80 degree C (+/-0.60 degree C 2 SD) and repeatability was +/-0.6 degree C. CONCLUSION: Although easy to use, infrared thermometry requires careful handling and experienced users. To get optimal recordings, the time between consecutive measurements should not be less than 90 seconds. Recordings should be taken immediately after positioning the device in the auditory canal. Best results are obtained when the grip of the device follows the ramus mandibulae like a telephone handle. The lower readings of the infrared thermometer compared to tympanic contact-probes indicate that the obtained readings represent the temperature of the auditory canal rather than of the tympanic membrane itself. PMID- 9340029 TI - [Effectiveness, side effects and costs of postoperative pain therapy: intravenous and epidural patient-controlled analgesia (PCA)]. AB - PURPOSE: Improvement of the quality of analgesia, reduction of side effects and costs by application of epidural (PCEA) in comparison to intravenous patient controlled analgesia (PCA) in postoperative pain treatment. METHODS: 62 patients with upper abdominal surgery took part in this randomised prospective study which was approved by the local ethics committee. Epidural catheters were inserted at T 8/9 (group PCEA). General anaesthesia was performed with propofol, sufentanil 2 micrograms/kg, pancuronium, enflurane and O2:N2O = 1:2. Postoperative analgesia consisted of epidural bupivacaine 0.25% + sufentanil 2 micrograms/ml (BS). (bolus 0.05 ml/kg, lockout 10 min) in group PCEA, or of intravenous morphine (bolus 2 mg. lockout 10 min) in group PCA. The following parameters were recorded until the evening of postoperative day 4: pain intensity at rest (VASR, 1-10) and on coughing (VASH, 1-10), blood pressure, heart rate, blood gas analysis, ability to ambulate, pruritus, nausea/vomiting (PONV), patient satisfaction (0-4), time and expenses for postoperative pain treatment. RESULTS: Median VASR (1 vs 2) and VASH (3 vs 4.5) were lower, cough intensity (2 vs 1) and patient satisfaction score (4 vs 3) were higher in PCEA compared to PCA. Ability to ambulate, pruritus, PONV, haemodynamics, paO2 and paCO2 were comparable. Postoperative pain treatment with PCEA was more time-consuming (407 vs 299 min) and expensive (71 vs 40 S/day) than PCA. CONCLUSION: PCEA in comparison to PCA after major abdominal surgery provides superior analgesia with comparable side effects at approximately 80% higher costs. PMID- 9340030 TI - [Effects of human i.v. immunoglobulin on bacterial clearance and granulocyte function in endotoxinemia]. AB - PURPOSE: The therapeutic impact of intravenous immunoglobulins (ivIG) in septic patients remains controversial. Until now, the mechanisms of action have not been fully elucidated. Since polymorphonuclear neutrophils (PMN) play a key role in host defence, this study focuses on the effects of ivIG on bacterial clearance and PMN respiratory burst activity during endotoxinaemia. For this purpose, it was investigated whether ivIG improves blood clearance and organ colonisation as well as PMN functions after experimentally induced bacteraemia in rabbits. METHODS: The experiments were performed in 30 anaesthetised rabbits. To determine quantification of bacterial killing in vivo, defined numbers of exogenous Escherichia (E.) coli 1.3 x 10(8) CFU) were injected intravenously in untreated animals (n = 10) or 60 min after infusion of endotoxin (LPS: 40 micrograms/kg/h) in groups without (n = 10), and after pretreatment with ivIG (Sandoglobulin, 0.5 g/kg body weight, n = 10), respectively. Parameters monitored were rates of bacterial elimination from the blood, LPS clearance, arterial pressure, blood gases and white blood cell counts, PMN burst activity was determined using a flow cytometry assay. Samples of liver, kidney, spleen and lung were collected for bacterial counts 180 min following E. coli injection. RESULTS: Compared to controls, endotoxinaemia resulted in a prolonged elimination of the injected E. coli out of the blood associated with a significantly (p < 0.01) higher colonisation of all organs. Pretreatment with ivIG improved LPS clearance and significantly reduced bacterial colonisation of lung and kidney (p < 0.01). This was paralleled by an enhanced PMN respiratory burst activity compared to untreated animals (p < 0.05). CONCLUSION: The reduced bacterial colonisation of lung and kidney in correlation with an increased PMN bactericidal activity in endotoxinaemia suggest an improved granulocyte-dependent bacterial killing due to ivIG application. PMID- 9340031 TI - [Consent and patient education in anesthesia. Legal principles and forensic consequences]. PMID- 9340032 TI - [Jugular bulb oximetry--a new neuro-anesthesiologic standard monitoring device? Contra]. PMID- 9340033 TI - [Jugular bulb oximetry--a new neuro-anesthesiologic standard monitoring device? Pro]. PMID- 9340034 TI - [Cerebral air embolism after central venous catheter]. AB - Introduction of air into the arterial circulation can cause cerebral air embolism, leading to severe neurological deficits. A case is reported on a patient suffering from fatal cerebral air embolism after a subclavian vein catheter had been inserted. The risks associated with inserting and removing central venous catheters are described. Apart from the pathogenesis of a paradoxical air embolism in a patient with a right-to-left shunt due to a patent foramen ovale, air embolism can occur if a large amount of air traverses the pulmonary circulation. The ability of the pulmonary vasculature to filter air may be exceeded by a bolus injection of more than 30 ml air. Air embolism is suspected if acute neurological symptoms occur after inserting a central venous catheter. Echocardiography, especially transoesophageal echocardiography. Is highly sensitive in detecting air emboli in the ventricles. Treatment is effected with hyperbaric oxygen and standard measures of intensive-care medicine. PMID- 9340036 TI - [Report on the 24th Neonatal and Infant Respiratory Symposium in Aspen, Colorado, March 1997]. PMID- 9340035 TI - [Asymmetric hypertrophic cardiomyopathy in a septic patient--intraoperative preliminary diagnosis with transesophageal echocardiography]. AB - Cardiac dysrhythmias in septic patients often reflect hypovolaemia, hypokalaemia or endocarditis as cardiac manifestation of the infection, but may be indicative for underlying cardiac pathology previously undiagnosed. We report on the case of a patient with severe peritonitis, on whom transoesophageal echocardiography (TEE) had been performed intraoperatively due to progressive circulatory instability. TEE revealed first diagnosis of asymmetric hypertrophic cardiomyopathy, which complicated the features of septic syndrome. Further perioperative treatment to support the circulation was successfully adjusted on the grounds of this diagnosis. PMID- 9340038 TI - [Has evolution made a mistake? Living with hemoglobin!]. PMID- 9340037 TI - [Comment on the Mini-Symposium, "Extreme anemia in transfusion refusal"]. PMID- 9340039 TI - International Symposium on Hepatology. Beijing, China, August 12-15, 1997. Abstracts. PMID- 9340040 TI - [Structure ond regulation of gene expression of tyrosine aminotransferase and proopiomelanocortin in mammals]. PMID- 9340041 TI - [Post-clinical period of disease: metabolic aspects]. AB - Examining the biochemical processes in the rehabilitative period after diseases of vascular and inflammatory genesis showed it expedient to identify an asymptomatic postclinical period of disease as an independent stage of metabolic remission. The guideline for assessing the completeness of metabolic processes was to study such indices reflecting the nonspecific pathological and etiological essence of diseases as erythrocytic phospholipids and fatty acids, as well as indices of the lipid peroxidation-antioxidative defense system, a lysosomal enzyme beta-glucosidase, uric acid and hydroxyproline. The duration of an asymptomatic postclinical stage was found to be determined by the time of completeness of metabolic processes and to be 3-12 months after the onset of clinical recovery. PMID- 9340042 TI - [Hemorheological profiles in patients with arterial hypertension and bronchial asthma]. AB - Hemorheological parameters were studied in patients with essential hypertension (men and women) and those with bronchial asthma. The rheological findings were established as a hemorheological profile. The latter is a set of macro- and microrheological parameters under various pathological conditions. The data show that blood viscosity was increased and correlate with blood pressure in hypertensive patients. There were more significant changes in the male populations of the two groups. The main cause of decreased blood fluidity and oxygen transport efficiency under these conditions is associated with hemoconcentration, elevated plasma levels of protein and increased plasma viscosity. The findings suggest that with the concept of a hemorheological profile one can correct rheological disturbances under various clinical conditions. PMID- 9340043 TI - [Changes in hemostasis and vagal tonus in healthy individuals and patients with facial neuropathy by micro-dose heparin stimulation of nasal receptors]. AB - The effects of heparin microdoses on vagal tone (the Bayevsky heart rate variability index) and blood coagulation time were examined in 15 healthy men and women and in 14 patients with acute facial neuropathy. Nasal inhalation of heparin solution vapour (thrice every 30 sec, interrupted by a 60-80 sec interval) without changes in olfactory sensation or nasal secretion prolongs the onset and completion of blood clotting and decreased heart rate (vagal activation) in normotony volunteers. The similar procedure of distilled water nasal inhalation appears to be ineffective. PMID- 9340044 TI - [Examination of vagus nerve tonus in various animal species]. AB - Experiments on pigeons, guinea pigs, rats, cats, rabbits, and dogs showed that tachycardia occurring after ligation of vagus nerves was caused by dissection of their inhibitory tone. They failed to confirm the hypothesis of the sympathetic origin of vagotomic tachycardia. The vagus nerve through the cholinergic neurons may exert heterodirectional (inhibitory and stimulating) effects on the performance of the heart only in animals with its tone. In the absence of tone, there may be only inhibition and stimulation was only short-term as a result of cessation of the inhibitory effect--postvagal tachycardia. The earlier established idea of the inhibitory tone of the vagus nerve remains valid. PMID- 9340045 TI - [Stability of epidemic process in context of biological system]. AB - A new hypothesis of formation (activation) of an epidemic process in the interseasonal and interepidemic periods at the expense of self-regulatory mechanisms associated with variations of parasitic biosystem diversity is suggested. The period of minimum intense morbidity in its multiannual and annual dynamics is shown to be the most vulnerable span of an epidemic process. PMID- 9340046 TI - [N.A. Iudaev scientist and founder of science]. PMID- 9340047 TI - [Role of bacterial lipopolysaccharides in human diseases: evolution of outlook]. AB - The authors propose a new conception of the role of bacterial lipopolysaccharides (LPS) in human illnesses. Being a structural component of a great variety of gram negative bacteria, upon the entrance of human body bacterial LPS serve as a biological signal indicative of bacterial invasion. This activates body defenses striving to limit the inflammatory focus, to arrest dissemination of the bacteria and their cleaning up. Such response of the body is feasible only in case of adequate function of the body defenses. Functional failure of the latter leads to inadequate response of the body which can not control systemic inflammation with polyorganic lesions. PMID- 9340048 TI - [On bioethical and legal studies of use of gene therapy]. AB - The bioethical and legal issues of research developments and the use of gene therapy in Russia are discussed. Prerequisites for development of gene therapy, the trends of researches and the national specific features of the application of gene therapy (religious confessions, ethnicity, health care service) are dealt with. PMID- 9340049 TI - [Biomedical ethics as a form of professional defense of physician's personality]. AB - The past 3 years have been marked by great changes in the sociohumatarian sector of medical education in Russia. They are associated with the introduction of biomedical ethics teaching. The Meeting on Philosophic Problems of Medicine (Academy of Medical Sciences, 1996) indicates that this subject is taught at the departments of philosophy of Russia's medical higher educational establishments. At the same time there are real trends for bringing biomedical ethics to medical law and for vanishing the former subject in some special subjects. The paper analyzes the reasons for these trends and concludes that with the introduction of new biomedical technologies and the new medical legislation, it is the significance of biomedical ethics as a regulator of physician-patient relations, as a form of protection of a physician's personality which especially increases, which requires its special and individual study and teaching. PMID- 9340050 TI - [The concept of sociomedical work in Russia in the late 1990]. AB - A concept of sociomedical work as a new kind of multiprofessional activities has been worked out. The concept has been adjusted to the present-day conditions of Russia and society. The specific features of national health care services and social protection of the country's population are taken into consideration as well. The chief points of the concept are as follows: the place and role of sociomedical work among other related activities in this field, the characteristics of the social sections of persons who need care, the basic direction of the work itself, the principles of organization of this kind of work, the main ways of its implementation, the basic points, principles, and criteria of the measures to be made, the solution of problems in the staff, and financing sources. PMID- 9340051 TI - [Main results of basic researches on complex problems of medicine in 1995]. PMID- 9340052 TI - [Role of extragonadal estrogens and hormonal carcinogenesis]. PMID- 9340053 TI - [Struchkov MV is an innovator in Russian surgery (to his 90th birthday)]. PMID- 9340054 TI - [Current problems of regulation of adrenocortical function]. AB - The currently available data on the regulation of corticosteroid secretion are analyzed. ACTH is the main regulator during acute stress. It accelerates the biosynthesis of glucocorticoids at the stage of cholesterol delivery to the mitochondrial inner membrane where the latter is converted to pregnenolone by side-chain cleavage. The Steroidogenic Acute Regulatory Protein (STAR) could play a great role in the supply of mitochondria with cholesterol. Under chronic stress, adrenal hypertrophy is largely involved in higher corticosteroid formation. Neuropeptides, cytokines, and growth factors modulate adrenal function. Prolactin which plays a role as a mitogenic factor in many tissues takes an active part in the control of adrenal function and proliferation. PMID- 9340055 TI - [Radiation pneumonitis]. PMID- 9340056 TI - [The value of diagnostic methods used to assess the response to treatment in patients with limited small cell lung carcinoma]. AB - Tumor response is used as a main criterion whether or not the treatment yields an anticancer activity. The tumor response criteria are defined by WHO recommendation but little is known about the tests must be used. The aim of this paper was to compare the degree of response to the treatment of 268 patients with limited small cell lung cancer, using independently 3 methods: radiological, bronchoscopic and cytological of bronchial material. Particular categories of response (CR, PR NR and presence or absence of carcinomatous cells) were related to survival time of patients independently to method of assessment. Multivarinte Cox analysis selected 3 parameters related to 3 different methods as independent survival risk factors. We conclude that each of diagnostic method (chest x-ray, bronchoscopy, cytological examination of bronchial material yield independent information correlated with survival risk of particular patient. PMID- 9340057 TI - [Chemotherapy versus chemoradiotherapy in patients with limited small cell lung carcinoma]. AB - 62 patients with a limited small cell lung cancer were randomly qualified into two groups. 32 patients of the first group were treated only with the chemotherapy regimen, consisted of three drugs (Carboplatine, Etoposide and Vincristine administered in 6 courses, on regular, 3-weeks basis). The second group of 30 patients had been treated with the identical chemotherapy schedule, but alternatively combined with a primary site irradiation in a total dose of 40Gy, applied in parts after the chemotherapy courses 2, 3, and 4. The significantly higher proportion of a complete remission results was observed in the alternate-treatment group: 14/30 (46.7%), compared with the chemotherapy-only group: 10/32 (31%). Alternate chemoradiotherapy resulted both in the increased median remission duration time, and the increased median survival time. Only in the alternate chemotherapy group, in 14/30 patients (46.7%) the pneumotoxicity symptoms appeared, whilst no differences in other organ-specific treatment induced toxic effects were noted. PMID- 9340058 TI - [Pulmonary mycobacteriosis--diagnostic problem and prevalence in Poland (a retrospective study)]. AB - The diagnosis of pulmonary mycobacteriosis was verified in 267 patients in Poland, classified in the years 1991-1995 to group VI M. Among 267 patients, 213 were men in age 28-83 years (median 43 years) and 54 women in age 24-81 years (median 50 years). The verification of the diagnosis was based on the authors own criteria of pulmonary mycobacteriosis and colonisation. These criteria included at least two positive sputum cultures and presence or absence of the respiratory tract inflammation. Pulmonary mycobacteriosis was recognised in 199 patients (168 men and 31 women). Colonisation was diagnosed in 41 patients (23 men and 18 women). Non-tuberculous mycobacteria most often responsible for pulmonary mycobacteriosis were identified as M. kansasii, M. avium--intracellulare and M. xenopi. Majority of patients with pulmonary mycobacteriosis or with colonisation had pulmonary lesions caused by tuberculosis in the past or suffered from chronic obstructive pulmonary disease. In 63 cases, data concerning previous diseases of the respiratory tract were not available. PMID- 9340059 TI - [Levels of endothelin-I in bronchoalveolar fluid of patients with selected respiratory tract diseases]. AB - Endothelin-I (ET-I) levels in BALF of symptomatic (n = 7) and asymptomatic (n = 10) asthmatic patients, sarcoidosis (n = 10), allergic alveolitis (n = 6) and healthy volunteers (n = 6) was evaluated. In all patients BALF level of endothelin-I was assessed by radioimmunoassay. We observed that patients with symptomatic asthma had more increased amounts of ET-I in BALF in comparison with asymptomatic asthmatics, patients with sarcoidosis, allergic alveolitis and control group. CONCLUSIONS: 1. Presence of ET-I in BALF indicates that this peptide is involved in the pathogenesis of bronchial asthma, sarcoidosis and allergic alveolitis. 2. Endothelin-I is involved in bronchial smooth muscle contraction. PMID- 9340060 TI - [Changes in lung parenchyma in high resolution computer tomography of patients with type type I sarcoidosis]. AB - The aim of this study was to describe the high-resolution computed tomography (HRCT) appearances of pulmonary abnormalities in 34 cases of type I sarcoidosis according to radiographic classification--lymphadenopathy only). Abnormalities were seen in 56% cases in which the pulmonary parenchyma appeared on radiographs. Nodules (1-4 mm) were seen in 41%. Other lesions as linear network were--present in 11.8% and ground-glass opacities--in 8.8%. The opacities are clustered in the peribronchovascular spaces, with relative sparing of the lung periphery and predominate in upper and middle lung zones. This study demonstrated the usefulness of HRCT in the correct diagnosis of type I sarcoidosis. PMID- 9340061 TI - [Levels of procollagen III in bronchial alveolar lavage fluid in patients with sarcoidosis]. AB - In 24 patients with sarcoidosis without pulmonary function disturbances significantly higher concentrations of procollagen III (PIIIP) in bronchoalveolar lavage fluid (BALF) were observed in comparison with control group. Patients in II radiological stage shower higher PIIIP concentrations in BALF than patients in I stage of sarcoidosis. PMID- 9340062 TI - [Soft tissue sarcomas as a rare cause of pleural effusion]. AB - Malignant disease is the cause of about 24% of all pleural effusions. They are caused mainly by lung and breast cancer. Three cases of pleural effusion caused by very rare neoplasm, soft tissues sarcoma are presented. In two of them the lesion found in the leg was observed for 4-14 month and not connected with the presence of pleural effusion. Difficulties in the histologic diagnosis of pleural sarcoma and of differentiating this tumour from mesothelioma are also presented. PMID- 9340063 TI - [In vitro susceptibility to antifungal agents of Candida strains isolated from patients with various diseases of the respiratory tract]. AB - The aim of the study was the estimation of in vitro susceptibility to antifungal agents of yeast isolated from sputum of 70 respiratory diseases patients using the disc-diffusion method-antimycogram. The following agents were tested: amphotericin B, 5-fluorocytosine, nystatin, ketoconazole, fluconazole. Only Candida strains were isolated from sputum, 82% of them were Candida albicans. We noted differences in susceptibility to antimycotics of Candida strains. The best antimycotic in vitro was 5-fluorocytosine. 54% of isolated Candida strains were resistant to 1 or more antimycotics. PMID- 9340064 TI - [The role of bacteriologic tests for diagnosis of tuberculosis in clinics other than tuberculosis control clinics (preliminary evaluation of test methods and their results)]. AB - Broadening of bacteriological testing for tubercle bacilli presence concerns patients with chronic cough treated in out-patients clinics and in hospital wards excluding tuberculosis control wards. Bacteriological tests have been carried out in tuberculosis laboratories as ordered by doctors of out-patient clinics and hospitals. From 05.11.1993 to 30.09.1994, 3917 tests were carried out on various specimens supplied to laboratories: sputum, urine, pleural fluid, peritoneal fluid, pus from lymph nodes. In 60 specimens positive results were obtained with the use of direct method or the mycobacterial culture. Pulmonary tuberculosis was diagnosed in 23 patients, renal tuberculosis--in 2 patients, tuberculosis of bones--in 1 case, tuberculosis of the peripheral lymph nodes--in 1 case, peritoneal tuberculosis--in 1 case. All the patients underwent specialized treatment. The above presented results justify the continuation of the tests. PMID- 9340065 TI - [Pleural empyema as a complication of descending necrotizing mediastinitis]. AB - Fulminant infections of oropharyngeal origin may cause the most lethal form of mediastinitis due to extension of the infection along the neck fascial planes or by lymphatic way: descending necrotizing mediastinitis. Two cases of this rare life-threatening entity of odontogenic and tonsillar origin are reported. A pleural empyema and pericarditis as complications were observed. Bacteriologic etiology, possibilities of the early diagnosis and the aggressive treatment by means of a thoracotomy are discussed. PMID- 9340066 TI - [Chylothorax in the course of squamous cell lung cancer]. AB - Chylothorax is a rare condition which might be caused by trauma of the chest, malignant tumors, congenital malformation, or inflammation of lymphatic nodes. It occurs as a complication of malignancy in about 50% of all cases, either as the first and the only symptom or at a very late stage of the disease. We presented a case of the 62 year old male with squamous cell cancer of the right lung, treated by chemo- and radiotherapy. Leakage of lymph to the pleura complicated the clinical status of the patient. Due to the primary disease the patient was treated symptomatically, in the hospital and later on in the out-patient clinic. Actually, eleven years after being diagnosed, despite massive fibrosis of the right lung the patient is doing relativity well, working part time. PMID- 9340067 TI - [Chlamydia pneumoniae as an etiologic agent in severe pneumonias]. AB - Two cases with pneumonia are presented. The diagnosis was based on the examination of the laryngeal secretions. The presence of inclusion bodies of Chlamydia pneumoniae were found in the Hep-2 cell culture. Treatment with doxycycline and erythromycin was effective. PMID- 9340068 TI - [Histiocytosis X--report of 3 cases with different types of lung changes]. AB - Three cases of histiocytosis X were described. Diagnosis was confirmed with histological examination of the lung taken during pleuroscopy in 2 cases and- with typical HRCT lung scan in 1 case. PMID- 9340069 TI - [Synchronous bilateral carcinoma of the lung cured with surgery and radiotherapy]. AB - A case of a primary synchronous bilateral lung cancer is presented. Tumor of the left lung was treated with lobectomy, tumor of the right lung with radiotherapy. Patient survived 5-years without evidence of disease recurrence. PMID- 9340071 TI - [Pneumocystis pneumonia in patients with steroid-dependent asthma]. PMID- 9340070 TI - [Immunologic reactions in the course of lung cancer]. PMID- 9340072 TI - [Bronchial carcinoid]. PMID- 9340073 TI - [Bronchoscopic cryotherapy]. PMID- 9340074 TI - Effect of absolute and relative gap sizes in visual search for closure. AB - In three visual search experiments using completely closed squares and squares with gaps, the physical gap size and the ratio of gap size relative to contour length were systematically manipulated. In search for an open square among closed squares, the search rates were fast and independent of the physical gap size or the gap-to-contour ratio. This is consistent with previous studies (e.g., Treisman & Souther, 1985). In contrast, in search for a closed square among open squares, the gap-to-contour ratio determined the search rates, irrespective of physical gap size. The latter result suggests that visual search is based not on local contour information but on global closure information, which seems to be constructed at some higher level of visual processing, probably at the level of perceptual representations of shapes and objects (e.g., Elder & Zucker, 1993; Rensink & Enns, 1995). PMID- 9340075 TI - Patterns of eye movements during parallel and serial visual search tasks. AB - Eye movements were monitored while subjects performed parallel and serial search tasks. In Experiment 1a, subjects searched for an "O" among "X"s (parallel condition) and for a "T" among "L"s (serial condition). In the parallel condition in Experiment 1b, "[symbol: see text]" was the target, and "O"s were distractors; in the serial condition these stimuli switched roles. Displays contained 1, 12, or 24 stimuli, with both target-present and target-absent trials. RT and eye movement measures (number of fixations, saccadic error, and latency to move) indicated that search efficiency was greatest in the parallel conditions, followed by the serial condition of experiment 1a and, finally, by the serial condition of Experiment 1b. This suggests that eye movements are correlated with the attentional processes underlying visual search. PMID- 9340076 TI - Stimulus modality and stimulus-response compatibility in absolute identification. AB - Accuracy and response time (RT) were measured in the absolute identification (AI) of 10 unidimensional perfectly pairwise discriminable stimuli. One group of 20 subjects performed a visual AI task involving line segments of variable length. A second group of 20 subjects participated in an auditory task with the stimuli composed of pure tones of variable intensity. Subjects performed the task under two conditions: a spatially compatible and a spatially incompatible stimulus response mapping. Results showed greater accuracy for the visual modality and longer RT for the incompatible mapping. The experimental factors did not substantially alter the bowing observed when performance was plotted according to the ordinal position of the stimuli. The data do not support the hypothesis that the bow effect is attributable to motor programming or motor adjustment stages. PMID- 9340077 TI - Translating between orthogonally oriented stimulus and response arrays in four choice reaction tasks. AB - Three experiments examined performance of four-choice reaction tasks using stimulus and response arrays oriented along parallel or orthogonal axes. All used a procedure in which pairs of locations were precued in advance of the target stimulus. Responses were slower for orthogonal than for parallel stimulus response sets, but the pattern of relative precuing benefits was similar. Complete transfer occurred when the stimulus array was changed from an orthogonal to a parallel orientation with respect to the response array after three sessions of practice. Transfer was also evident when the orientation of the response array was changed from orthogonal to parallel with respect to the stimulus array, as long as the assignment of stimulus locations to fingers was not altered. The results suggest that coding in the four-choice task is by relative location regardless of whether the stimulus and response sets are oriented orthogonally, and that an additional transformation operation to align the frames of reference is performed for orthogonal orientations. PMID- 9340078 TI - Interkey timing in piano performance and typing. AB - In typing, when the fingers executing two successive movements are on the same hand, the time between keystrokes is longer than when the fingers are on different hands. Biomechanical limitations of the hands are thought to account for this difference. The generality of this finding was explored by investigating skilled pianists' performance of two successive notes. Experiment 1 failed to find comparable differences in timing as a function of the hands involved. Experiment 2, employing both a piano production and a typing task, replicated the previous piano performance results, and revealed that the timing differences in typing were limited to letter sequences requiring fore-aft and lateral finger movements. Experiment 3 extended this finding to piano performance. Together, these findings clarify the nature of biomechanical constraints on skilled manual performance. PMID- 9340079 TI - Proceedings of the 25th scientific meeting of the Group of Chromatography and Related Techniques of the Spanish Royal Society of Chemistry. Barcelona, Spain, 22-25 October 1996. PMID- 9340080 TI - [Powder-coated gloves. The problem has not yet been recognized]. PMID- 9340081 TI - [Complications and false diagnoses caused by powder residues. Powder-coated gloves should be banned from medical daily life]. PMID- 9340082 TI - Human T-cell receptor zeta chain gene Map position 1q23.1. PMID- 9340083 TI - Proteinase inhibitor 8 Map position 18q21.3. PMID- 9340084 TI - [Polymerase chain reaction in the diagnosis of tuberculosis]. PMID- 9340085 TI - [Lipoproteins as functional associates of protein-lipid complexes (literature review)]. PMID- 9340086 TI - [Diagnostic significance of glycolipid sialic acids in hyperlipoproteinemias]. AB - Total and lipid-bound sialic acids in the blood were measured and lipoprotein metabolism assessed in 219 men and women with primary (n = 129, coronary patients) and secondary (n = 66, patients with type II diabetes) hyperlipoproteinemias and normolipidemia (n = 24, normal subjects). The level of total sialic acids in coronary patients reliably differed from that in normal subjects or diabetics. The level of lipid-bound sialic acids was the same in coronary patients and normal subjects and reliably higher in diabetics. The results do not confirm the hypothesis about the probable diagnostic value of total and lipid-bound sialic acids as markers of coronary disease and coronary atherosclerosis. Measurement of total sialic acids may be one of the tests verifying the disorders of lipoprotein metabolism. PMID- 9340087 TI - [Follow-up of fibrin clot growth in blood plasma]. AB - The authors propose a device for following up the kinetics of the fibrin clot enlargement in the plasma. Coagulation is induced by contact activation with a fragment of an arterial wall put into the plasma. Besides the initial time of clot formation, the device helps assess the rate of clot growth. Two phases of contact-induced clotting may be distinguished. The first is slow activation of the clotting system and formation of minor amounts of fibrin, the other is rapid growth of the clot, with the linear increment of the clot size being constant during 15 min. PMID- 9340088 TI - [Use of flow cytofluorimetry in the analysis of interactions of mononuclear phagocytes with protein antigens]. AB - Flow cytofluorometry of samples stained with fluorochrome-labeled antigen may be used to study the interactions between human blood calls and antigens. Using fluorescein isothiocyanate-labeled tuberculin (after the original method), the authors found that human blood monocytes actively bind labeled tuberculin. Studies of the concentrations from 0.1 to 12,500 ng/ml showed that saturation of the cell capacity for endocytosis is attained at certain doses, after which their surface label prevails. The share of tuberculin-binding lymphocytes was appreciably lower than that of monocytes at the same doses of the antigen. Clinical application of the method for assessing the antigen-presenting function of macrophages is discussed. PMID- 9340089 TI - [Bacterial vaginosis]. PMID- 9340090 TI - [Apoptosis: an important stage in the assessment of the immune system according to the pathogenetic principle]. PMID- 9340092 TI - [Comparative analysis of 2 sanitary-bacteriological methods of detecting bacteria of the Escherichia coli group in the environment]. AB - A total of 965 parallel tests for coli bacilli were carried out using the wash out or print methods in the course of sanitary and epidemiologic inspection of different objects. The methods used in the study were equally sensitive and provided compatible results which were in good correlation. The print method with bacterial tests is more rapid (by 24 hours) and helps detect other bacteria which grow in Endo's medium; moreover, it is three times cheaper than the labor consuming wash-out method. PMID- 9340091 TI - [Immunoenzyme analysis in the determination of antibodies to main classes of cell phospholipids in human blood]. AB - A modified ELISA is proposed for detecting antiphospholipid antibodies of the IgM and IgG isotypes, including antibodies to cardiolipin, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine, and sphingomyelin in human blood serum. The degree of phospholipid saturation, its phase (lamellar or hexagonal), purity, oxidation index of the phospholipid preparation, and conditions of solid phase treatment are important factors providing a sufficiently high sensitivity and reliability of measurements. The clinical value of the method was demonstrated in examinations of 70 patients with habitual abortions. The method can be used in studies of the spectrum of antiphospholipid antibodies in different diseases involving an increase of their blood serum level. PMID- 9340094 TI - [Culture medium for isolation of causative agent of diphtheria]. PMID- 9340093 TI - [Method of polymerase chain reaction in laboratory practice]. PMID- 9340096 TI - [Development and application of software for clinical-laboratory diagnosis]. PMID- 9340095 TI - [Evaluation of the sensitivity and specificity of methods of rapid diagnosis in bacterial vaginosis]. AB - Bacterial vaginosis was diagnosed in 35.8% of 109 patients of a reproductive age complaining of discharge from the genital tract. In 33% of cases bacterial vaginosis was associated with vaginal candidiasis. The most sensitive and specific method of rapid diagnosis of bacterial vaginosis is microscopic examination of Gram-stained smears, which may be used as a diagnostic method, whose sensitivity and specificity are almost 100%. Such tests as pH-metry, amine test, "key" cells, and assessment of vaginal discharge (normal or abnormal), should be used in complex. They may be performed by the therapist screening patients to detect bacterial vaginosis. PMID- 9340097 TI - [Unit for examining an intact blood smear. Osmotic resistance of erythrocytes]. PMID- 9340098 TI - [Economics of quality control in clinical laboratories]. PMID- 9340099 TI - [Diagnosis of chlamydial and gonococcal infection using the polymerase chain reaction method]. PMID- 9340101 TI - Annual meeting of the Hungarian Society for Microbiology. Nyiregyhaza, August 21 23, 1996. Abstracts. PMID- 9340100 TI - [Method of polymerase chain reaction in laboratory diagnosis of hepatitis B and C]. PMID- 9340102 TI - On: follicular transplantation by Bernstein and Rassman. PMID- 9340103 TI - Management of acute gastrointestinal hemorrhage in anticoagulated patients. AB - Bleeding is the major risk associated with anticoagulation therapy. The gastrointestinal tract is the most common site of bleeding. Anticoagulated patients who present with acute gastrointestinal hemorrhage pose great therapeutic challenges. In patients who experience a life-threatening hemorrhage, difficult decisions must be made regarding reversal of anticoagulation, timing of endoscopy and endoscopic therapy, and when to reinstitute anticoagulation. The current literature on the approach to patients who present with major gastrointestinal bleeding while on anticoagulant therapy is reviewed. PMID- 9340104 TI - [Results of sampling of the population on the development of a healthy life style]. PMID- 9340105 TI - [Social problems of families having a child with infantile cerebral palsy]. PMID- 9340106 TI - [Medico-social problems of young pregnant women]. AB - A total of 255 women aged under 17 and 320 pregnant women aged 18 were followed up in 1994. Medicosocial aspects of pregnancy were investigated using special questionnaires, medical aspects were analyzed using medical files. This enabled us to characterize from medical and social viewpoints young women, parent families, and socio-hygienic factors of pregnancy. PMID- 9340107 TI - [Fundamentals of syphilis prevention today]. PMID- 9340108 TI - [Role of the general practitioner in the organization of the primary component of medical care]. AB - A program "Introduction of General Practitioners (Family Doctors) in the Primary Health Care System" has been developed in the Samara region. The first results are presented. PMID- 9340109 TI - [Results of an experiment on the transition to the family practice service]. AB - The activity of physicians of general profile over 2 months was analyzed in the course of experiment on transfer from the local therapeutic service to general medical practice. The results of general practitioners' work were found compatible with those of local therapeutic service. PMID- 9340110 TI - [Cost of outpatient care]. AB - Presents data on distribution of finances in public health institutions for physicians of different specialization, engaged in outpatient consultations, and analyzes the values for institutions of different types. The results may be used for economic analysis of the activities of public health institutions. PMID- 9340111 TI - [Optimizing the model of developing general practitioner service]. PMID- 9340112 TI - [Assessing the effectiveness and quality of state epidemiologic surveillance and the activities of the first level sanitary epidemiologic institutions]. AB - Validates a complex of methodological approaches to assessing the efficiency of sanitary epidemiological institutions today. Shows the specificities of such assessment for the institutions of the first-level management. Validates a demonstration model for computer processing of data on the efficiency of departments of environmental hygiene and of specialists in this field and recommends this model for regional centers of State Sanitary and Epidemiological Surveillance. PMID- 9340113 TI - [The integral bed fund of a territory]. PMID- 9340114 TI - Analysis of the activity of therapeutic and prophylactic hospitals for validating the structural and functional reorganization of public health of a territory (as exemplified by the Tambov district). AB - Analyzes the activities of inpatient treatment-and-prophylaxis institutions in the Tambov region of Russia. Shows the good reserves for improving the efficacy of hospitals, consisting in more accurate assessment of the bed funds and rational distribution of medical and paramedical staff in public health institutions. PMID- 9340115 TI - [Automated information system for management of an outpatient hospital]. AB - The development of the organization and functional model of an outpatient hospital of a therapeutic profile functioning as a department of an outpatient clinic prompted the author to create an automated information system for the collection, analysis, and storage of medical information using personal computers. This will make medicostatistical information available for heads of institutions and departments and offer variants of management decisions on improving the management and function of therapeutic outpatient hospitals. PMID- 9340116 TI - [The content analysis of medical sociological information]. AB - Discusses the application of content analysis of medical periodicals as exemplified by neurosurgery. Content analysis is an obligatory element of modern research in public health organization. PMID- 9340117 TI - [Organization of geriatric care in the Samara region under conditions of obligatory medical insurance]. AB - The authors share the experience they gained in rendering medical care to elderly and senile patients and describe the specific features of such care under conditions of obligatory medical insurance. The study was carried out in the Samara region. PMID- 9340118 TI - [Training of public health organizers]. AB - This paper continues the discussion of problems in the training and activities of public health organizers (No. 2, 1996). The author confirms that measures are to be taken by the Ministry of Health which should be aimed at fortifying the role and authority of Social Medicine and Public Health Management Departments in the reformation and management of public health. A system of public health organizers is to be created in the Russian Federation, which is to include improved training of physicians on the whole, resident training, postgraduate training on a full time basis and by correspondence, and updating cycles. The author emphasizes that a physician without special training cannot become a public health manager or member of governing body. PMID- 9340119 TI - [Knowledge control in training in social medicine]. PMID- 9340120 TI - [The problem of licensing healers and ensuring social security of patients]. AB - Development of marketing in public health, cases of poor care of the patients, inadequate legal basis for relations between healers and the state organs responsible for the quality of medical care result in an increase of the number of popular healers, both real and false. The author emphasizes the need of creating as quickly as possible the laws ensuring social protection of a patient from incompetent treatment. PMID- 9340121 TI - [Reform of university medical education in Austria in the second half of the XVIII century]. PMID- 9340122 TI - [The 150th anniversary of using narcosis in Russia]. PMID- 9340123 TI - [The Moscow physician and historian of medicine, R. A. Stanelli--author of books on Paracelsus]. PMID- 9340125 TI - A proposal to improve quality, increase efficiency, and expand access in the U.S. health care system. Board of Directors of the American Medical Informatics Association. PMID- 9340124 TI - [Medico-demographic parameters and standard of living of the Russian population]. AB - Correlation-regression analysis of medical demographic characteristics, parameters of the economic status of the country, and level of life in 1990-1994 showed a marked correlation between a drop of the level of life in Russia and deterioration of the medical demographic situation: decrease of birth rate, increase of mortality, and shortening of the expected life span. The most significant factors were decrease of the total gross product in % of 1990, decrease of the number of staff of industrial enterprises, and a lower consumption of meat and milk products in kg per capita. A single-factor exponential model proved to be the most adequate. The derived equations may be used for predicting the medical demographic parameters for Russia using the detected factors. PMID- 9340126 TI - Paclitaxel package insert. PMID- 9340127 TI - [The trauma patient: open hospital choice?]. AB - Victims of trauma have usually no choice regarding the physician or hospital they are admitted to. In order to deliver the best possible trauma care it is crucial that trauma victims first receive competent on site primary care before being admitted directly to a hospital that is sufficiently equipped and qualified to take care of their injuries. Recent literature suggests that individual outcomes, but also per-case costs of trauma patients clearly improve, when prehospital care, triage and admission to specially designed trauma centres are coordinated within regional trauma systems. This provides supporting evidence for the recent proposals by the Swiss Medical Association (FMH) in 1996 who formulated 12 statements regarding rescue services in Switzerland. In order to optimise rescue and trauma care there is an urgent need for restructuring existing systems nation wide. Trauma patients may thus to some degree lose their freedom in choosing their preferred physician or hospital. PMID- 9340128 TI - [Traffic accidents from the social medicine viewpoint--a medical history connection with present time]. AB - A historically excursus seems sometimes useful to overlook actually problems in medicine. The example of the railway accidents shows the problems of the medical appraisal with regard to the consequences of an accident. After initiation of a third party insurance in Germany 1871 with reference to the new means of transportation a lot of people tried to get a compensation after imaginary or real injury following a railway accident. Very soon physicians had to give an expert's opinion on the complaints of those concerned. 1879 Rigler and 1918 Horn published monographs regarding this problem. Apart of real impairments very often complaints had no organic reason. Oppenheim introduced 1889 the concept of "traumatic neurosis". The scientific discussion remained in the following decades and finally it was rejected until to our times. Today the terms "posttraumatic stress disorder", "psychogenic pain disorder" or "adjustment disorder" are valid. In present times the problem of the whiplash injuries takes the place of the railway accidents. Here also are either organic reasons discussed or psychogenic reactions presumed. The modern techniques in neuroimaging and neuropsychology are inconsistent and the results not generally accepted. In the individual case they are often little helpful. The medicolegal problems are still important and often reason for prolonged forensic confrontations. The discussions more than 100 years ago seem nowadays still actual. The terms have changed, the problems remain. PMID- 9340130 TI - [Are complications in cruciate ligament replacement operations with patellar tendon transplantation dependent on surgical technique and surgical timing?]. AB - HYPOTHESIS: In a retrospective study we analyzed our results of ACL reconstructions with a patellar tendon graft. We wanted to know if the complications were dependent upon timing and technique of surgery. METHOD: We reviewed 283 patients after ACL-reconstruction, who underwent an operation with bone patellar tendon graft between 1984-1993. In our study we particularly looked for complications. The overall rate of complications was 21.6% dependent on the applied technique. Infections, DVTs, limitations of movement and graft failures were the most common complications. Furthermore we analyzed the timing of operation. Arthrofribrosis was less common in the group with delayed reconstruction (6.1%) whereas in the primary reconstruction group the rate was 17.6%. For this reason we changed our management with regard to the timing of operation. Meniscal injuries were the most common additional injuries in both groups. Conservatively treated ACL-ruptures showed a high rate of mensical ruptures in combination with cartilage injuries. CONCLUSION: Because of these results we put more emphasis on patient information to achieve the optimal result and to meet the individual needs for every patient. PMID- 9340129 TI - [Intermediate term results after arthroscopic meniscus suture]. AB - QUESTION: Do results after arthroscopic meniscal repair justify the bigger expenditure and the longer rehabilitation opposite to the arthroscopic menisectomy? METHOD: An arthroscopic meniscal repair in inside-out technique was carried out by the same surgeon in 35 consecutive patients with meniscal tears (tears at the meniscocapsular junction with best prognosis after repair are not subject of this study). Rehabilitation was standardized. After a mean follow-up time of 43 months, 27 (77%) of these patients, 12 without and 15 with an additional anterior cruciate ligament (ACL) reconstruction, were evaluated personally. RESULTS: There were 4 (14.8%) reruptures (3 true reruptures, 1 "rerupture" out of the repair side) and I lesion of the N. saphenus with limited dysaesthesia. Three of the 4 reruptures occurred in patients with isolated meniscal repair, only one in patients with additional ACL-reconstruction. The functional results (Lysholm-score, knee mobility) and the subjective result were good in patients without reruptures. Postoperatively, patients were unable for work and sports for a long time, but these times being shorter for isolated meniscal repairs (5.4 and 12 weeks respectively) than for patients with an additional ACL-reconstruction (12 and 29 weeks respectively). CONCLUSIONS: In terms of knee function and osteoarthrosis, arthroscopic meniscal repair has better results than partial meniscectomy and should be preferred, therefore, especially in young patients. A sufficient number of meniscus sutures should be performed and existing ligament instabilities should be repaired. A good rehabilitation program is essential. Efforts for a better acceptance of the method in patients and family doctors are necessary. PMID- 9340131 TI - [Treatment of fresh Tossy III acromioclavicular joint dislocation by ligament suture and temporary fixation with the clavicular hooked plate]. AB - PURPOSE: Retrospective analysis of the functional and radiographic outcome of a new temporary fixation device in the surgical treatment of acromioclavicular joint dislocation stage Tossy III. MATERIAL AND METHODS: Between 1986 and 1993, 19 patients with acute acromioclavicular (AC) dislocation (stage Tossy III) were treated with the clavicle hook-plate as a temporary fixation device combined with suture of the ligaments. Active shoulder mobilization started three days postoperatively. The mean follow-up was 3.9 years (range, 2 to 9.5 years). RESULTS: The mean Constant Score was 94 points (range, 83 to 100 points). All of the patients were able to resume their occupational activities and seventeen (89.5%) could completely resume their prior program of physical fitness. Radiological signs of an AC arthritis were found in 21% of the cases, but only 1 patient had a painful AC-joint. The average distance between the superior aspect of the coracoid process and the inferior aspect of the clavicle in the stress x ray (5 kp each side) of both AC-joints was preoperatively 20.7 mm (range, 12-50 mm) comparing to 9.5 mm (range, 6-15 mm) for the operated shoulder and to 9 mm (range, 4 to 12 mm) for the nonoperated side in the follow-up. No reluxation was found. Two superficial wound infections (10.5%) were treated successfully with antibiotics without removal of the implant. Neither breakage nor loosening of the clavicle hook-plate were observed. CONCLUSION: Our mid- and longterm results confirm the value of the clavicle hook-plate as an alternative to other temporary fixation devices in the surgical treatment of acromioclavicular dislocation stage Tossy III. PMID- 9340133 TI - [Isolated dislocation of the elbow]. AB - GOALS OF THE STUDY: To analyse the results of non surgical management of isolated dislocation (without any fractures) of the elbow in adult. MATERIAL AND METHODS: We reviewed with an average follow up of 36 months 22 patients (22 elbows) who had been treated conservatively after a first episode of posterior dislocation of the elbow. RESULTS: Nineteen patients (86%) reported an excellent or a good subjective result, although 64% suffered occasionally from pain. None had presented a recurrent dislocation. At physical examination, 27% had a restricted extension of the elbow of 10 degrees and more, and 27% had some kind of chronic laxity. Fifty percent had modifications visible on the X-rays. We found no correlation between laxity and duration of immobilization. At contrary, patients who were immobilized for a longer time than 3 weeks suffered more often of a painful restricted extension of the elbow. DISCUSSION: Conservative management of posterior dislocation of the elbow has a good prognosis. Occasional pain and chronic laxity are often present but well supported. Early mobilization decreases the risk of permanent limited extension of the elbow. PMID- 9340132 TI - [Elbow fractures in elderly patients]. AB - Are the well established methods for operative treatment of fractures about the elbow and their results also applicable to elderly people with osteoporosis? In a retrospective study 23 of 32 patients with 33 operatively stabilized fractures of the distal humerus and the proximal radius and ulna, could be reviewed and personally questioned. The fracture type, the complications and the range of motion of the elbow were analysed and classified. Over 75% of the cases presented with a good to excellent range of motion with no increase of complications. 80% of the patients were satisfied with their result after surgical treatment of the fracture. Fractures of the elbow in elderly people with osteoporosis may be sucessfuly treated by ORIF. It appears important to start early with postoperative physiotherapy. Thereby good functional results can be expected without an increase of complications. PMID- 9340134 TI - [Blunt thoracic trauma with bronchial rupture]. AB - Diagnosis of bronchial rupture after blunt chest trauma may be difficult and this is demonstrated on behalf of four patients treated in our institution. Bronchoscopy is mandatory for exclusion or confirmation of a bronchial rupture. In addition spiral CT scan was found to be helpful for diagnosis and localisation of bronchial injury. Early diagnosis allows prompt surgical therapy that will avoid irreversible loss of pulmonary parenchyma. PMID- 9340136 TI - Sarcopenia and physical performance in old age. Proceedings of a workshop. Bethesda, Maryland, July 9-10, 1996. PMID- 9340135 TI - [Blunt abdominal trauma with lesion of the abdominal aorta--a case report]. AB - A 58-year old lady, involved in a head-on motor vehicle crash suffered a severe intestinal injury associated with an intimal flap lesion of the distal abdominal aorta. Thrombotic occlusion of the aortic bifurcation with clinical evidence of lower extremity ischemia was noted. The management of blunt injury to the abdominal aorta is discussed with special regard to placing prosthetic material in a potentially infected field. PMID- 9340137 TI - [A study of bilingual Galician-Castillian aphasic patients]. AB - INTRODUCTION: Aphasia in bilingual persons has some striking peculiarities. Most studies have been carried out in mono-lingual societies, where the second language has usually been learnt after immigration. In Galicia, linguistic closeness and simultaneous learning of Galician and Castilian languages permits the testing of neuropsychological theories conceived in other circumstances. OBJECTIVE: In aphasic bilingual Galician-Castilian patients we studied how this affected denomination, designation and translation, different modes of recovery, variables associated with differential affectation, the presence and influence of specific phenomena of aphasia in bilingual persons on changes in dominance and mixing of the languages. Material and methods. We selected 49 patients, 29 men and 20 women aged between 32 and 85 years old. We analyzed the variables associated with differential affectation: types of aphasia, size and site of the lesion, aetiology, age, educational level, and the presence and influence of phenomena specific to aphasia in bilingual persons. RESULTS: The test most affected was translation, to a lesser extent denomination, and finally designation. Capacity for recovery is greater in the dominant language. Translation recovers better than denomination and this better than designation. We found no changes in dominance, no selective loss of either language but have seen some interference in the non-dominant language. CONCLUSIONS: The two languages are seen to be recovered equally, as were other languages previously studied in aphasiology. PMID- 9340138 TI - [Histological prediction of cerebral tumors using SPECT with 201T1]. AB - INTRODUCTION AND OBJECTIVE: SPECT with 201Tl provides information regarding the degree of malignancy of cerebral tumours, their possible relapses, the differentiation of necrotic tissue in the tumours following chemotherapy or radiotherapy and permits differentiation into zones of various grades of histological malignancy. MATERIAL AND METHODS: We carried out a prospective analysis of the usefulness of SPECT with 201Tl for the histological prognosis of cerebral tumours. For one year 68 patients diagnosed (on CT and/or MR) as having an expansive cerebral lesion were studied. The early uptake (ICP) and retention (R) indices were calculated, and these results correlated with the morbid anatomy (AP) findings and the results obtained with surgery and stereotactic biopsy. Four patients were excluded due to lack of AP results. RESULTS: Significant differences were found between the ICP of grade I-II astrocytomas (1.34 +/- 0.52) and glioblastomas multiformes (2.56 +/- 0.57), between the ICP of meningiomas (4.53 +/- 1.68) and metastases (2.45 +/- 0.58) and between those of meningiomas and all glial tumours. With regard to IR, we saw significant differences between the figures for meningiomas (0.63 +/- 0.13) and meningiomas with malignant relapses (0.94 +/- 0.17) and between metastases (0.8 +/- 0.03) and all glial tumours. CONCLUSIONS: From our study, it may be concluded that rapid, high captation of 201Tl (high ICP) with a slow fall (high IR), is associated with a process showing malignancy on histological study (malignant relapse of meningioma, glioblastoma multiforme, metastasis), whilst high take-up (high ICP) with rapid elimination (low IR) corresponds to a benign hypervascularized tumour (meningioma). PMID- 9340139 TI - [Insomnia and somnolence in epilepsy]. AB - INTRODUCTION AND METHODS: Based on the well-known association between sleep disorders and epilepsy which share common functional elements, we investigated the existence of sleep disorders in general as seen in a series of patients who attended the electrophysiology clinic, using a simple questionnaire method dealing with sleep disorders and somnolence. RESULTS: It was seen that there was a markedly greater prevalence of sleep disorders in epileptics as compared with non-epileptic controls. It was also observed that in partial crises these disorders are more important, mainly because of the global sleep deficit and increased nocturnal waking. Control of the crises was also important since patients with long periods free of critical episodes had fewer disorders than those with poor control. CONCLUSIONS: The discrepancy between clinical critical control and sleep anomalies may be explained by the subclinical effect (which is usually underestimated) of inter-ictal discharges. PMID- 9340140 TI - [Validity and reliability of the migraine self-questionnaire Alcoi-1995]. AB - INTRODUCTION: In epidemiological investigation valid, reliable means of diagnosing the illnesses studies are essential. OBJECTIVE: To validate the self questionnaire 'Alcoi 1995' for the diagnosis of migraine. METHODS: We used the questionnaire Alcoi-1992 (which had been validated previously) to find the prevalence of migraine in homogeneous populations. After this, we selected the significant items to obtain a shorter, more specific questionnaire for diagnosis of migraine, which we called 'Alcoi 1995'. We selected a population of patients and healthy volunteers. The diagnosis of migraine (neurologist and self questionnaire) was made on criteria of the International Headache Society (IHS, 1988). RESULTS: There were 71 persons included in the study, 40 patients (5 men and 35 women those average age was 39.7 +/- 14.4 years) and 31 healthy volunteers (15 men and 16 women with an average age of 36 +/- 14.4). The sensitivity, specificity, positive and negative predictive value, and the Kappa concordance index for migraine (100%, 100%, 100%, 100% and 1); migraine with no aura (75%, 100%, 100%, 86% and 0.79) and migraine with a typical aura (100%, 88%, 63%, 100% and 0.71) were calculated. CONCLUSIONS: Our results show a high degree of validity and reliability when the self-questionnaire is used for the diagnosis of migraine. It is a tool for use in epidemiological studies which is quick and easy to use, and is cheap. PMID- 9340141 TI - [Prevalence of migraine in a homogeneous population using the self-questionnaire 'Alcoi-1995']. AB - INTRODUCTION: There are numerous studies of the prevalence of migraine and very varied findings. Amongst the many reasons for this would seem to be the use of questionnaires for screening populations, absent or incorrect validation of these questionnaires and the use of impracticable diagnostic criteria. OBJECTIVE: To determine the prevalence of migraine in a homogeneous population using the self questionnaire 'Alcoi 1995' which had previously been validated for the diagnosis of migraine. A self-questionnaire was given to all workers in the same company. Diagnostic criteria of the International Headache Society (IHS, 1988) were used for diagnosis of migraine. RESULTS: The study included 140 persons (43.6% men and 56.4% women with an average age of 26.7 +/- 4.8 years). There was a prevalence of headache and migraine (IC 95%): 90.8% (75.01-100) and 24.3% (16.3-32.3) respectively. The sex prevalence of headache and migraine was 88.5% (88.3-88.8) and 11.5% (3.4-19.4) for men and 34.2% (21.5-47) for women respectively. CONCLUSIONS: There are a large number of studies of the prevalence of migraine with wide variability in the figures obtained (1-2 at 35%). These variations depend on the population studied, age and sex sampled, diagnostic criteria used and the method employed in the survey. After validation of the questionnaire for diagnosis of migraine, this should be used in homogeneous populations before use in broad population studies. Our findings support the use of a suitably validated questionnaire as a useful method for the diagnosis of migraine in epidemiological studies. PMID- 9340142 TI - [The combined application of visual evoked potentials and the quantified electroencephalogram improve discrimination of cerebral maturity]. AB - INTRODUCTION AND OBJECTIVE: The aim of this job is to evaluate brain maturation by means of Electroencephalogram (EEG) and Visual Evoked Potentials stimulated with flash (VEP-flash) quantitative analysis techniques. MATERIAL AND METHODS: The transversal study is made on a sample of 96 subjects in which EEG and VEP flash, first isolated and then joining both, are analyzed. The selection of spectral parameters was done taking care of all the subjects were selected in the sense of maximizing brain maturation discrimination. Multivariate analysis techniques for classifying subjects were used. EEG and VEP-flash variables were selected with the linear discriminant analysis. In the EEG case the variables take into account, as a reference, either the median of the power spectrum or either the time instant in which the spectral power in every band reaches its maximum value. In the joined EEG-VEP-flash the VEP variables which give more information were related with the slopes and distances between the basic peaks of the evoked response (N1, P1 and N2) and age. For brain maturation evaluation the variables in the occipital channels are sufficient, being those of the right hemisphere the most diagnostic significative ones. CONCLUSION: The joined use of EEG and VEP-flash means an improvement in the maturative level discrimination regarding to the isolated consideration of any of them. Variables obtained from the EEG-VEP-flash are enough for brain maturation evaluation. PMID- 9340143 TI - [Perinatal differences in asphyxic full-term newborns in relation to the presence of hypoxic-ischemic encephalopathy]. AB - OBJECTIVE: To study the perinatal differences of full-term newborn infants with perinatal asphyxia in relation to their neurologic manifestations (postasphyctic encephalopathy). MATERIAL AND METHODS: Prospective epidemiologic study over perinatal asphyxia in full-term infants born in our hospital between november 1991-february 1995. Perinatal asphyxia was graded as non-severe (first minute Apgar score < or = 6 and/or umbilical artery pH < 7.20, with abnormal fetal heart rate patterns and/or meconium stained amniotic fluid, and the need for immediate neonatal resuscitation) and severe (first minute Apgar score < or = 3 and umbilical artery pH < 7.10). Hypoxic-ischemic encephalopathy was graded as mild, moderate and severe based on classification of Levene and Sarnat & Sarnat. The perinatal variables were graded as prenatal (gestationals and obstetrics), neonatal (resuscitation, general data of the newborn, and organic manifestations of asphyxia) and postneonatal (neurologic sequelae at follow-up). RESULTS: During the study period there were 3,342 full-term, live births. Perinatal asphyxia developed in 156 (31 severe and 125 non-severe). Neurologic manifestations were present in 25.6% of asphyxiated newborns: 40 cases of hypoxic-ischemic encephalopathy (mild in 30, moderate in 5 and severe in 5). The main differences between asphyxiated newborns with and without hypoxic-ischemic encephalopathy were: chronic maternal diseases, pathologic obstetric antecedents, distocic deliveries, neonatal resuscitation, severity of perinatal asphyxia, sex and weight of newborns, days and origin of admission, birth injury, extraneurologic manifestations (mainly pulmonary, digestive, haemodynamic and cardiologic) during neonatal period, and neurologic sequelae at follow-up. CONCLUSIONS: The main perinatal differences in relation to postasphyctic encephalopathy are important in the knowledgement of their pathogenic mechanism (interrelation between neurologic and extraneurologic manifestations) and their follow-up (hypoxic ischemic encephalopathy is the most important prognostic factor of neurologic sequelae in the full-term asphyctic newborn). PMID- 9340144 TI - [Neuropathy associated with arteriosclerosis]. AB - OBJECTIVE: To determine the frequency and characteristics of the neuropathy associated with arteriosclerosis. MATERIAL AND METHODS: A prospective clinical and electrophysiological study was made of 29 male patients with arteriosclerosis, in whom other causes of polyneuropathy had been excluded. RESULTS: Eleven patients complained of paresthesiae (mostly mild). In 11 patients there were signs of polyneuropathy on clinical examination. Neurophysiological studies were abnormal in 11 patients, suggesting the presence of predominantly sensitive axonal neuropathy. In five patients with paresthesiae both physical examination and electrophysiological studies were normal. In 17 patients there were changes in the somatosensory evoked potentials. The brainstem auditory evoked potentials of 27 patients were suggestive of diffuse changes in central nervous conduction, together with super-imposed focal lesions. There were no differences as regards age, signs of disease in the legs or of the involvement of widespread illness, whether they were smokers, ex-smokers or non-smokers, the number of cigarettes smoked daily or the total duration of the smoking habit between the patients with and without clinical or electrophysiological polyneuropathy. CONCLUSIONS: Approximately one third of the patients with arteriosclerosis have clinical or electrophysiological signs suggestive of predominantly sensitive axonal polyneuropathy. In some cases the patients had paresthesia but no changes were seen on physical or electrophysiological examination. The evoked potentials showed diffuse changes in central nervous conduction, and in some cases this was associated with signs of focal lesions. PMID- 9340145 TI - [Toward the federation of neurological journals in Spanish?]. PMID- 9340146 TI - [The role of documentation services in scientific production]. PMID- 9340147 TI - [The Internet list of Neurology in Spanish]. PMID- 9340148 TI - [School achievement and epilepsy]. PMID- 9340149 TI - [Myelographic diagnosis of an epidural arterio-venous fistula]. AB - INTRODUCTION: Epidural arterio-venous fistulae are a little known clinical condition. They are probably commoner than is thought, since diagnosis and angiographic demonstration are difficult. CLINICAL CASE: We present the case of a 49 year old man with the clinical features of chronic myelo-radiculopathy with episodes of intermittent medullary claudication. This shows the use of myelography, which in certain cases allows clear detection of vascular structures which might otherwise pass unnoticed. CONCLUSIONS: One should think of this condition when compatible features are found and there is no other diagnosis, since if confirmed by myelography and/or magnetic resonance prior to angiographic study, it may benefit from embolization, which is an effective treatment for this. PMID- 9340150 TI - [Unilateral agenesis of the internal carotid artery in childhood: description of a case]. AB - INTRODUCTION: Unilateral or bilateral agenesis of the internal carotid artery has not often been described in children since it is usually asymptomatic. CLINICAL CASE: We describe a case of isolated unilateral agenesis of the internal carotid artery in a child of 20 months old who presented with two episodes of status convulsives, the first after a head injury and the second when he was febrile. The anomaly was a casual finding, diagnosed on cerebral magnetic resonance when there was no signal corresponding to carotid blood flow. It was confirmed by echography-doppler and angiographic magnetic resonance. We also describe the neuro-radiological findings which led to the diagnosis of this anomaly. CONCLUSIONS: We consider that unilateral agenesis of the carotid artery was a casual finding in this case. The provisional diagnosis may be made when there is absence of vascular signals on conventional magnetic resonance. PMID- 9340151 TI - [Benign myoclonic epilepsy in childhood. A case report]. AB - INTRODUCTION: Benign myoclonic epilepsy of childhood is a rare syndrome which appears at between 4 months and 3 years of age. The prognosis is good if diagnosed and treated early. It is characterized by many short crises (usually of 3 seconds and not more than 5-10 seconds long), proximal and cephalic jerking movements without falling to the ground, and at no particular time of the day. In the EEG polygraph background activity continues and crises coincide with generalized spike and wave or multiple spike and wave discharges of 1 to 2 seconds, accompanied by isolated myoclonic movements in the neck and deltoid muscles, which persist during NREM sleep. Benign epilepsy of childhood usually responds to monotherapy with valproic acid. In our case photosensitivity appeared at 7 years of age with persistence of generalized spikes and waves during sleep. CONCLUSION: We suggest that photosensitivity may be used as an index of the clinical course, and that treatment should continue to be given until photosensitivity disappears. PMID- 9340152 TI - [Familial strio-pallido++-dentate calcification]. AB - INTRODUCTION: Calcification of the dentate nucleus of the cerebellum may be seen on cerebral CT in 0.3-0.5% of patients with no symptoms or extra-pyramidal signs. Although there are many causes, some cases show a family incidence. Clinical cases. We present two cases, mother and son, in whom the predominant clinical findings were poor language and a bilateral extrapyramidal syndrome. Laboratory tests, hormone and immunological studies were normal. In both patients cerebral CT showed bilateral calcification of the basal nuclei and dentate nuclei of the cerebellum. DISCUSSION: Our patients fulfilled the clinical and neuro-imaging criteria described in familial strio-pallido-dentate calcification. There is no relationship between the duration and intensity of symptoms, and the extension of the calcification. However, all patients with calcification show clinical features. The predominant features are poor language and a bilateral extra pyramidal syndrome. These observations indicate that in patients with calcification of the basal ganglia and dentate nuclei of the cerebellum it is necessary to study first degree relatives in order to identify the condition. PMID- 9340153 TI - [Diagnosis of centronuclear myopathy in adults]. AB - INTRODUCTION: Centro-nuclear myopathy is a congenital myopathy characterized by the presence of central nuclei on muscle biopsy. Three clinical forms have been distinguished. Classification depends on the type inherited, age of onset and degree of muscle involvement. CLINICAL CASE: We describe the case of a female patient in whom the diagnosis of centro-nuclear myopathy was made at the age of 53. The patient had not been studied previously, but was sent to us by the Department of Anaesthesia. The clinical features had first appeared in infancy. There was no family history of this disorder. Apparently this was a sporadic case. CONCLUSIONS: In the differential diagnosis of adult patients with girdle paresthesias centro-nuclear myopathy should be included. This unusual muscle disorder may be need to be considered if anaesthesia is required. PMID- 9340154 TI - [Deafness secondary to lumbar puncture]. AB - INTRODUCTION AND OBJECTIVE: Disorders of hearing have been described in patients who have undergone lumbar puncture. The lower frequencies are most affected, temporarily and with spontaneous recovery in most cases. It would seem that the cochlea aqueduct (AC) (an anatomical structure found in the internal ear) is the part involved in the pathogenesis of this infrequent complication. The object of this paper is to review the different hypothesis put forward by various authors on the subject, emphasizing the one which seems most probable. At the same time, we offer a new vision of this little known and often forgotten anatomical structure of the internal ear. The anatomy and physiology of the AC is considered, studying the part played by the labyrinth fluid when the patient undergoes an operation involving the cerebro-spinal fluid (CSF) or the subarachnoid space. CONCLUSIONS: Whenever this technique is to be used, it is essential to obtain a clinical history to find out whether the patient has, or has had, any problems of hearing, and if so of what type. All patients should be warned of the possibility of deafness, in most cases temporary, and if the patient has hydrops endolymphaticus (syndrome of Meniere) of the high risk of worsening his hearing threshold. We therefore recommend precise evaluation of the use of this technique. PMID- 9340155 TI - [Topiramate, a new antiepileptic drug]. AB - OBJECTIVE: The characteristics of the new anti-epileptic drug topiramato (TPM) are described: multiple mechanism of action, favourable pharmaco-kinetics, wide therapeutic range and is relatively well tolerated, Development. Its initial usefulness in patients with partial crises resistant to other anti-epileptic drugs has been shown. TPM is equally effective in primary generalized crises, and in patients with the Lennox-Gastaut syndrome, in whom no problem of tolerance has been seen. CONCLUSION: TPM is a valuable drug for the treatment of epilepsy. PMID- 9340156 TI - [Factors to consider in the evaluation of intellectual achievement of an epileptic child]. AB - In this paper, factors related to the intellectual achievement of epileptic children are reviewed. These should be taken into account by medical and paramedical staff when making the intellectual prognosis of each patient. The factors related to epilepsy include: The type of crisis and epileptic syndrome, the aetiology of the epilepsy, genetic factors, age of onset, duration of the disorder and frequency of crises, electroencephalographic alterations, a history of status epilepticus, the antiepileptic drugs and their metabolic effects. The most important psychosocial factors are: The school, the family and the patient's personality. The significance of the multifactorial effects is emphasized. PMID- 9340157 TI - [The effect of melatonin as anti-convulsant and neuron protector]. AB - INTRODUCTION: Melatonin is the principal hormone secreted by the pineal gland. In human beings the pineal gland is found on the posterior aspect of the midbrain. Melatonin is synthesized from tryptophan following a circadian rhythm, with low levels during the day and high levels during the night. It regulates many biological processes in relation to the cycles of light and darkness. DEVELOPMENT: Its first known function was that of inducing sleep. In experimental animals its effect as a depressor of the central nervous system and its anti convulsive action have been shown. Few studies have been done in human beings, although there is some evidence of its beneficial effect in epileptic patients, improving both the frequency of the crises and the EEG tracing. In our experience we gave melatonin to a girl with severe myoclonic epilepsy which did not respond to usual treatment, obtaining improvement in both the number of crises daily and in psycho-motor development. Several possible modes of action have been described for melatonin; increase in Gabaergic transmission at a cerebral level, where GABA is the main inhibitory neurotransmitter; interaction with benzodiazepinic cerebral receptors; it may owe its effect to the activity of its metabolites, particularly kinurenic and kinurenic acid; it may induce hormone changes in the organism. Recent studies show the marked anti-oxidant activity of melatonin. Its considerable capacity to accept free radicles resulting from biological processes has been shown and it thus acts as a cell protector. CONCLUSIONS: It seems reasonable to assume that melatonin has anticonvulsant and neuroprotector properties. Further study may define its pharmacological usefulness in these disorders. PMID- 9340158 TI - [Nicardipine and its indications in neurology]. AB - OBJECTIVE: Although the use of calcium-channel blockers is strongly supported by experimental data, its clinical application is far less clear. In this paper we intend to review the actual guidelines for using a member of this family of drugs, nicardipine. MATERIAL AND METHODS: We have carried out an intensive search for any clinical assay published with nicardipine all over the last years in different clinical entities such as subarachnoid haemorrhage, migraine, cerebrovascular pathology, etc. In many of them no clinical assays have been done or either they have too many methodological problems so that appropriate conclusions can be drawn. However most of them offer results which should have prompted the design of wider works in number of patients and follow-up periods which could offer conclusive answers to an important number of questions which remain open about the use of this class of drugs. CONCLUSIONS: We have only found a clear indication for the use of nicardipine in migraine and subarachnoid haemorrhage. In the rest of clinical entities reviewed we have found some experimental evidences but no clinical data to justify this use. It is highly recommendable that well designed clinical assays with an adequate number of patients are started in this field. PMID- 9340159 TI - [Memory and epilepsy]. AB - INTRODUCTION: The neuropsychological literature has shown the presence of memory deficits in patients with epilepsy. These alterations in memory are due to several factors, such as the aetiology of the epilepsy, age at onset of the seizures, the duration of the disorder, and the frequency and type of the epileptic seizures. DEVELOPMENT: These memory defects reflect the structural and functional damage caused by the repeated epileptic seizures and/or the anti epileptic drugs. Apart from this, neuro-surgical treatment of temporal lobe epilepsy may also lead to memory defects. These will depend on the site and side of the epileptic focus, the level of memory prior to surgery, the age of onset of the epileptic seizures, the chronological age at the time of operation and the reduction in the number of seizures after surgery. In spite of the evidence that memory is affected in patients with epilepsy, this does not mean that all people with epilepsy have these changes. CONCLUSIONS: Neuropsychological examination is of fundamental clinical importance since it allows the evaluation of cognitive function in each patient, and thus permits optimization of both the pharmacological treatment and intellectual and physical performance. PMID- 9340160 TI - [Status epilepticus]. AB - INTRODUCTION: Status epilepticus (SE) is a condition in which epileptic discharges are sufficiently prolonged or repeated so as to cause persistent changes in neurological function. DEVELOPMENT: At the present time the classification of SE seems to be incomplete and therefore provisional. However the broad subdivision into convulsive SE and non-convulsive SE is still useful in clinical practice when emergency treatment is required. The neuropathological changes are largely determined by the nature (excitory or inhibitory) and the duration of the event which originated the crisis. Tonic-clonic SE is only one of many variants of SE. However, unlike many others, it is a medical emergency which if not suitably treated may lead to permanent neurological damage. Neurone damage and death are the result of a series of parallel processes occurring at a systemic and neuronal level. Outstanding amongst the latter is the activation of a complex neurotoxic cascade. Better understanding of the physiopathological mechanisms occurring in SE would set the stage for a more logical use of treatment. CONCLUSIONS: With advances in the treatment of SE in recent years the prognosis of these patients has improved considerably. However, we should now consider the use of conventional anticonvulsant drugs alone to be insufficient, and that this should give way to polytherapy with neuroprotector agents. PMID- 9340161 TI - [Neuropsychological deterioration in Huntington's disease]. AB - INTRODUCTION: Huntington's disease (HD) is a neurodegenerative illness, autosomal dominantly inherited, that produces characteristic involuntary muscle movements (chorea) with cognitive abnormalities and personality. DEVELOPMENT: Neuropathological studies report premature cell death principally affecting the basal ganglia. The disease most commonly begins in the fourth or fifth decade of life, although in some cases, abnormalities have been documented in early childhood and as late as 50 years of age. Early cognitive deterioration in HD is not uniform, and patients with recently diagnosed HD may evidence selective deficits in learning and visuospatial/visuomotor ability, along with a relatively consistent pattern of impairment on subtests of the Wechsler Adult Intelligence Scale (WAIS). Neuroimaging reports of basal ganglia function have described striatal abnormality in HD and related with neuropsychological impairment. CONCLUSIONS: This paper provides the clinical-phenomenologic definition of the neuropsychological impairment and its associated features related with its neuropathological significance and to review the subject in the light of present day reports. PMID- 9340162 TI - [Language and aphasias]. AB - INTRODUCTION AND OBJECTIVE: Approximately 400,000 years ago men started to use language. Initially it was probably poor with few phonemes. With social evolution it became more complex, with the appearance of new phonemes and a more complete grammatical structure. The current concept of the processing of language dates, with little change, from the nineteenth century. DEVELOPMENT: With the birth of phrenology language began to be studied. This lead to the hypothesis of Wernicke, with two main areas joined by the fasciculo arcuato, which is still held to be valid with modifications by Gerchwind and Damasio, amongst others. Important advances in the study of language are due to Chomsky and his transformational grammar. This supports the universal structure of language, since one learns it following genetically determined laws. Language has three main aspects: creativity which makes both the transmitter and the receiver active participants in communication, the form from which words are constructed and the content of the message. Aphasia is an alteration in the comprehension and understanding of language, which may be the clinical expression of many different aetiologies. They help us to localize the lesion topographically. They are divided depending on the clinical signs, into motor or Broca's aphasia, in which understanding is conserved but the patient uses a language with poor grammatical structure, although the semantic content is acceptable: sensitive or Wernicke's aphasia, with inability to understand and language which is fluid but unintelligible; conduction aphasia due to limitation in the transmission of impulses from Wernicke's area to that of Broca, with acceptable understanding and fluid language and the trans-cortical aphasias where the main characteristic is indemnity of the capacity of repetition. CONCLUSIONS: The aphasias, as the expression of an alteration of language are an important support in the topographical localization of lesions, even before these can be shown on computerized tomography. PMID- 9340163 TI - [The etiology of neurological complications after cardiopulmonary bypass surgery]. AB - INTRODUCTION: Cardiopulmonary bypass (CEC) in the surgical treatment of cardiac diseases may cause the appearance of neurological damage of an intensity which varies between minor neuropsychological disorders and global cerebral anoxia. There are two mechanisms for the production of these lesions: ischaemic and embolic. The mortality associated with this type of complication is low, but morbidity may be considerable. The neurological disorders derived from CEC may be classified according to the aetiology and clinical findings. In the first group are included: severe cerebral anoxia, embolic cerebro-vascular accidents, microvascular embolias, lesions of spinal vascularization and lesions of the peripheral nerves. In the second group are: encephalic focal lesions, convulsive crises, lesions of the extra-pyramidal system, alterations in the level of consciousness and neuropsychological disorders. METHODS: Quantification of neuronal damage has been attempted by: monitoring cerebral blood flow and neurone metabolism, EEG and study of intra-operative evoked potentials, echography of the carotid, cardiac and ascending aorta, transcranial doppler, fluorescein angiography and the study of biochemical markers of neuronal and glial damage. Different studies have identified a series of factors which potentiate the risk of neurological lesions following CEC. These are: age, severe carotid disease, aortic atherosclerosis and previous cerebro-vascular haemorrhage, amongst others. An attempt is made to reduce the incidence of neurological complications by: pre operative evaluation of carotid bruits, hypothermia, careful surgical technique and the use of drugs with a neuroglial protector effect. None of these methods gives sufficiently effective protection to the central nervous system subjected to the changes involved in the use of CEC. CONCLUSION: There are still many unknown aspects of neurone pathology in these circumstances, leaving a door open to investigation. PMID- 9340164 TI - [Genetic predisposition to Alzheimer's disease]. AB - INTRODUCTION: Alzheimer's disease can present either as an early-onset or senile dementia. Its ethiology is heterogeneous but with common clinical and pathological features. DEVELOPMENT: Alzheimer's syndrome can occur with familial clustering. Some families with pre-senile dementia have shown three different genes which are associated with the pathological features. These genes carry on several mutations which have an autosomal dominant transmission: each mutation seems to be able to cause the pathological changes. The senile dementias are more common but dominant transmission has not been shown. However, risk genetic factors can play a rol. The epsilon 4 allel for apolipoprotein E has been clearly identified. CONCLUSIONS: It would be possible to use genetic tests to predict the appearance of presenile dementias but these tests are not available for the more common senile types of Alzheimer's dementia. PMID- 9340165 TI - [Neurology in Internet]. AB - OBJECTIVE: The quantity and complexity of the information to be found on the Internet makes anyone who has dared to navigate on the Net feel perplexed. To access the Net without a predetermined route to follow may prove to be a waste of time and very exasperating, as a tremendous amount of completely irrelevant information is discovered. The objective of this article is to give the reader of the REVISTA DE NEUROLOGIA an informative guide to help find useful information on the Net, and a classified list of potentially useful Internet addresses. DEVELOPMENT: We have analyzed the concept of 'search tool', and the types and characteristics of these. Specific search tools (both English and Spanish) which are currently available, are considered. We have searched the Net using various tool (Yahoo, Lycos, Altavista, Trobador and Ole) to find relevant information for the terms 'neurosciences' and 'neurology', and to verify the relevant interest of the sites on the Net thus encountered. CONCLUSIONS: We present the URL of 50 search tools and the 244 sites on the Net related to Neurology and the Neurosciences, classified according to the entity or group to which they correspond, and their ambit (Latin American/Spanish vs others). A short list of those which may be of most interest is also included. PMID- 9340166 TI - [Two literary characters in the language of neurology: Pickwick (1)]. PMID- 9340167 TI - [The Valencia Neurological Society. A 45-year-old man with epileptic crises]. PMID- 9340169 TI - [Chemoreceptors and the heart]. PMID- 9340168 TI - The legal implications of the veterinarian's role as a private practitioner and health professional, with particular reference to the human-animal bond: Part 1, The law of negligence. PMID- 9340170 TI - [Sympathetic-vagal euqilibrium and physical exercise]. PMID- 9340171 TI - [The impact of clinical trials in arterial hypertension in clinical practice]. PMID- 9340172 TI - [Clinical relevance of ischemic preconditioning]. PMID- 9340173 TI - [Secondary prevention of stroke through arterial blood pressure reduction]. AB - Large scale observational studies have conclusively demonstrated that systolic and diastolic blood pressure values are linearly related to the incidence of cerebrovascular diseases and that high blood pressure is an important risk factor for both primary and secondary development of stroke. Interventional studies have shown that blood pressure lowering by antihypertensive treatment reduces the incidence of stroke in hypertensive patients without a history of previous stroke. Whether this is the case also for the secondary prevention of cerebral ischemic attacks has not been unequivocally shown, however. The PROGRESS ("Perindopril Protection Against Recurrent Stroke Study") study has been designed and is under way to collect information on this important issue of the antihypertensive treatment, its purpose being to evaluate the blood pressure lowering effects with an ACE-inhibitor on recurrent stroke in an overall population of 6000 patients with a positive history of previous cerebral ischemic attacks or stroke. PMID- 9340174 TI - [Vasovagal syncope and increased baroreceptor activity: evidence for increased sensibility of the baroreflex and its rapid reversal with acute administration of metoprolol]. AB - In 17 patients suffering from recurrent episodes of vasovagal syncope as well as in 21 healthy subjects without clinical episodes of presyncope or syncope, we evaluated the reflex decrease in heart rate evoked by the phenilephrine test. In the syncopal patients, the measurements were taken 4-12 hours after the clinical appearance of syncope. We divided the syncopal patients as follows: 9 patients, undergoing pharmacological treatment, and 8 untreated patients (drug free arm). In the pharmacological arm of the study, an alternate, randomized administration of metoprolol (150 mg twice daily for 2 days) and verapamil (80 mg every 6 hours for 2 days) was provided. Therefore, in the pharmacological arm as well as in drug free patients, we tested again the baroreflex sensitivity, by means of iv phenilephrine bolus, 3 and 7 days after the clinical appearance of the syncopal event. The baroreflex sensitivity values were significantly higher in the syncopal group compared to the control group (21 +/- 5 vs 13 +/- 4.5 ms/mm Hg; p < 0.01). Of the two tested drugs, only the metoprolol produced a fast (day 3) decrease in baroreflex sensitivity. On the basis of measurements taken after 7 days, we noted a pattern of widespread reduction in baroreflex sensitivity values, found in both treated and untreated patients. In conclusion, patients with vasovagal syncope exhibited a more pronounced maximal parasympathetic activation compared to the control group. The high baroreflex sensitivity values were soon (day 3) reduced by metoprolol, but not by verapamil therapy; a spontaneous normalization in baroreflex sensitivity values was found 7 days after the clinical episode, regardless of therapy. PMID- 9340175 TI - [Circadian variation of sudden cardiac death in young people with and without coronary disease]. AB - Sudden cardiac death is not well known and provoking factors are yet mainly unknown. To clarify whether sudden cardiac death has a circadian rhythm in young people we have studied 40 patients < 45 years who died in Brescia between 1984 and 1993 of sudden cardiac death showing at autopsy features of coronary artery disease (CAD) and 12 patients aged < 30 years who died of sudden cardiac death without autoptic features of CAD. We observed a circadian rhythm in the hours of the morning in the two groups, more evident in patients without CAD. In patients with autoptic features of CAD, we also observed a higher rate of events during the winter months. We would like to stress the importance of the adrenergic system as a trigger able to produce the event. We believe that the role of the sympathetic nervous system is more important than other risk factors (for example platelet aggregability and blood viscosity) to precipitate sudden cardiac death, mainly because the circadian rhythm was more evident in patients without CAD. An increase of the data-base and a more detailed analysis of subgroups is necessary if we concretely want to prevent sudden cardiac death fitting antiarrhythmic therapy with circadian distribution of major events. We underline the practical impact of "chronorisk" together with the other cardiovascular risk factors. PMID- 9340177 TI - [Double-chamber right ventricle: a case report]. AB - The double-chamber right ventricle is a congenital cardiac malformation usually associated with other cardiac defects, seldom isolated and in adult subject. It is characterized by the presence of an anomalous bundle that divides the right ventricle into two chambers. The clinical and electrocardiographic signs of isolated double-chamber right ventricle are few and not specific. An echocardiographic diagnosis of isolated double-chamber right ventricle is reported. In a 18-year-old asymptomatic male with systolic murmur 2/6 at third space over the left sternal border, right ventricular hypertrophy and intraventricular conduction delay at ECG, two-dimensional echo showed an anomalous transversal muscle bundle that divided the right ventricle into two chambers, superior and inferior. Color Doppler showed a diastolic tricuspidal like flow through a paraseptal discontinuity of the bundle and a systolic jet that reached the right atrium, with a pressure gradient of 30.9 mmHg. The absence of symptoms and other cardiopathy, without significant right outflow tract obstruction, was considered as an index of a good prognosis; therefore cardiac catheterization was not advised. PMID- 9340176 TI - [Coronary angioplasty in acute myocardial infarction]. AB - Primary percutaneous transluminal coronary angioplasty (PTCA) for acute myocardial infarction (AMI) allows to obtain a higher reperfusion rate in the culprit vessel than thrombolytic therapy, reducing the incidence of death, non fatal reinfarction and recurrent ischemia. The aim of this study was to test the in-hospital and mid-term results of an early invasive strategy with PTCA in patients with AMI. Thirty-four patients with AMI underwent coronary angiography within 3 hours from the onset of symptoms. Twenty-four patients had anterior AMI and 3 were in cardiogenic shock. Three patients, 1 without significant lesions and 2 with multivessel diffuse coronary disease, were left out of the procedure, and 31 patients underwent PTCA. Twenty-six lesions were total occlusions with TIMI flow 0.A TIMI flow 1 was present in the other 5 vessels. Stent deployment was decided for 16 lesions (52%). Primary success (TIMI flow 3 with mean residual stenosis of 15 +/- 20%) was obtained in 30 patients (97%). In 1 patient recanalization of the anterior descending coronary artery was not possible due to tortuosity of the abdominal and thoracic aorta. At pre-discharge angiography a good result was confirmed in 24/25 patients. After 6 months only 1 patient (3%) underwent a new PTCA for recurrent angina. In conclusion, primary PTCA for AMI within 3 hours of symptom onset allows good in-hospital and mid-term results with a low rate of complications. PMID- 9340178 TI - [Percutaneous stenting of the renal arteries]. PMID- 9340179 TI - [A three-year long follow-up of a group of "everyday patients" in a mental health center]. AB - OBJECTIVE: This research concerns those patients who most attend the community Mental Health Centre (CSM), hereby called "everyday patients". According to a previous research (Pileggi et al., 1992) a sample of patients mostly attending the Centre had been pointed out. Basing on the number of attendances, it emerged that some of those patients (57) had been on the average attending the Centre more than twice a week and, despite being only 10% of the total number of users, they had taken on a large share of the services offered by the Centre (30%). Therefore, those patients were the ones the Centre had been working for more intensely and continuously. Three years later, the object of this research is to check the assistance and clinical destiny of such patients and compare their patterns of attendances to those ones regarding the remainder of the CSM users. The hypothesis is that "everyday patients" are assisted by different and continuous treatments and that such a procedure prevents patients from dropping out and determines a strong reduction in relapses and less frequent attendances. DESIGN: Longitudinal study on a 42 patient sample (19 males, 23 females) on therapy at CSM. SETTING: Mental Health Centre, "Saragozza" District, Sanitary Unit of Bologna. MAIN OUTCOME MEASURES: The following elements have been examined: 1) social and demographic features; 2) duration of psychiatric history; 3) clinical diagnosis according to DSM-III-R, set by patients personal psychiatrist; 4) global functioning level as examined by two psychiatrists or psychologists from the Centre, using DSM-III-R Global Functioning Scale (GFS); 5) actions carried out and patterns of using the CSM services over the past 12 months. Concordance measures among independent examiners (Cohen K) and non parametric variability measures for comparison between groups (Chi-square and Kruskal-Wallis tests) have been used. RESULTS AND CONCLUSION: Results partially confirm the original hypotheses. In particular, complicated services (psychological and pharmacological therapies and rehabilitation) are carried out for the most of "everyday patients" and much more intensely to them than to the remainder of the users. No drop-out has been found out, the global functioning level of the patients is good in most of cases and the number of necessary admissions to psychiatric wards has been reduced. However, the "attendance share" relevant to the sample of "everyday patients" is still high compared to the total number of the CSM users. Besides, discharge rate is nought. PMID- 9340180 TI - [Parental bonding as a predictive factor for the development of adult psychiatric disorders]. AB - OBJECTIVE: To asses the capacity of the Parental Bonding Instrument (PBI) to discriminate between normal subjects and clinical samples and between with different psychiatric diagnosis. DESIGN: The present paper analyzes the studies published between 1979 and 1995, which have used the PBI in normal subjects and clinical samples and have reported the respective means and standard deviations obtained on the two PBI dimensions: affection and control. Multiple comparisons were carried out between the mean scores of affection and control of: 1) samples with the same psychiatric diagnosis (intragroup comparison); 2) samples with different psychiatric diagnoses (intergroup comparison); 3) normal subjects and clinical samples. RESULTS: Of the 46 studies with normal and clinical subjects, 23 studies were selected for the analysis, reporting means and standard deviations and specifying the diagnostic criteria. Samples with the same psychiatric diagnosis had similar affection and control scores. With the exception of bipolar affective disorders and avoidant personality disorders, the prevalent parental style was for all diagnostic groups the affectionless control style. Within the affectionless control style, the PBI discriminated between panic attacks, borderline personality and drug addiction but not between schizophrenia, unipolar depression and anxiety disorder. The PBI discriminated also between normal subjects samples and samples with anxiety disorder, schizophrenia, bipolar affective disorder, personality disorder and drug addiction respectively. CONCLUSION: The results confirm previous suggestions from single studies that the perceived parental style as measured by the PBI can be considered a good predictor for the presence of psychiatric disorders excluding panic attacks, avoidant personality disorders and unipolar affective disorders. Although the different diagnostic groups do not differ in their perceived parental style (affectionless control), significant differences between some diagnostic groups within this category suggest that the PBI might have some specificity as well. PMID- 9340182 TI - [Guidelines for the preparation of CV and list of publications. To standardize the format for encouraging a more homogeneous evaluation of candidates for positions in psychiatry]. PMID- 9340181 TI - [Costs of psychiatric services: a study performed at a public mental center in the Lombardy Region, Italy]. AB - OBJECTIVE: The implementation of a simple methodology to estimate full costs of services provided by a public mental health centre. SETTING: CPS (NHS Mental Centre) Ussl 35, Magenta, Lombardy Region. METHOD: To estimate full costs of 16 types of service we followed a two step procedure. The first step was to estimate all costs attributable to the CPS. In the second one, we allocated this estimate to each type of service provided. We attributed to the CPS the following cost items: personnel, utilities (telephone, electricity, water, heating and cleaning), land & building, transports (for services provided outside the clinic) and a share of general cost of the USSL to which the CPS belongs. Full cost of each service was then calculated on the base of the yearly number of services provided and the time spent by each health professional. RESULTS: In 1995, the CPS provided 14,562 services. Total costs amounted to L 1,356 million, and more than three quarters of this amount was attributable to the personnel working at the CPS. Unit costs ranged from L 5,300 (drug administration) to L 442,400 (family therapy involving two professionals for 90 minutes) The unit cost of psychiatric visits, psychologist consultations and nurse domiciliary visits were L 105,300, L 106,600 and L 78,000, respectively. CONCLUSIONS: This approach requires accessible data and is relatively simple to manage. Some refinements are required, especially to improve the methodology for the determination and the allocation of overheads. However, we are convinced that this cost accounting procedure provides acceptable estimates of the services provided by the CPS. These estimates suggest that charges to be used to fund NHS providers may be too low, especially if fee-for-service will be the main funding source. PMID- 9340183 TI - [Psychotherapy research: why is it neglected in Italy?]. PMID- 9340184 TI - [Re-examining the hedgehogs anecdote]. PMID- 9340185 TI - [The cognitive role of epidemiology and its pitfalls]. AB - One of the most important roles of epidemiology is to seek causes of diseases. The causality in medicine is a difficult philosophical category which requires deep ontological thinking and a thorough methodological preparation. If this condition is not met, various misunderstandings and errors develop with serious consequences for clinical practice. In recent medical literature we find many controversial views which are abused by the press and cause mistrust of the public to medical science and attract patients to the dubious practices of alternative medicine. It is therefore the duty of research workers to pay more attention to modern epidemiological methods and their philosophy. The predominating pluricausal model of risk factors is recently criticized because of its excessive simplicity. reductionism and dualism which separates health from disease. The need arises of a new universal model of health where the dominant position will be held by social environmental, behavioral and psychological aspects. Modern epidemiology needs not only a correct methodology but also a proper theory. PMID- 9340186 TI - [Serotonin and treatment of mental disorders. Present status and future perspectives]. AB - Serotoninergic system is involved in the regulation of diverse biological and psychological functions and a variety of serotonin receptor subtypes represent a possible target for a new generation of medications. 5-HT receptors play an important role in both schizophrenia and depression. Modern strategies for treating schizophrenia profit from the existence of interaction between serotonin and dopamine systems. New drugs called serotonin-dopamine antagonists (SDAs) offer wider spectra of activity and lower extrapyramidal side effects liability. The principle of the SDAs is that the drug should be a potent serotonin 5-HT 2A antagonist, with slightly less potent dopamine D2 receptor-blocking properties. New pharmacological agents with great therapeutic potential and fewer side effects were recently developed also for the treatment of depression. Among these new antidepressives the serotonin selective reuptake inhibitors (SSRIs) currently play the most important role. PMID- 9340187 TI - [Metabolic sequelae of pancreatic transplantation using two different surgical techniques]. AB - BACKGROUND: By transplantation of the pancreas in diabetics type 1 long-term-term independence on exogenous insulin can be achieved. The extent of normalization of the carbohydrate metabolism can depend on the applied surgical technique. The objective of the submitted work was to compare indicators of compensation of diabetes one year after combined transplantation of the kidney and pancreas, using the method of transplantation of a segment of the pancreas with obliteration of the pancreatic duct by a polymer and the method of transplantation of the whole pancreas with drainage of the pancreatic duct into the urinary bladder. METHODS AND RESULTS: The authors examined two groups of recipients, 13 subjects each with full function of the pancreatic graft one year after transplantation where a combined transplantation of the kidney and pancreatic segment (group SP) had been performed or of the kidney and whole pancreas (group CP). The authors investigated the blood sugar level, glycated haemoglobin, intravenous glucose tolerance test, free insulin level and C-peptide as well as some indicators of the lipid metabolism and acid base balance. In both groups normal blood sugar levels were achieved, though the mean values in the course of the day were higher in group SP than in group CP (mean +/- SE 5.48 +/- 0.11 as compared with 4.98 +/- 0.09; p < 0.01). Glycated haemoglobin declined in group SP from the pretransplantation value of 9.31 +/- 0.09 to 6.40 +/- 0.10% and in group CP from 9.49 +/- 0.15 to 4.92 +/- 0.08%. In group CP the glycated haemoglobin after transplantation was significantly lower (p < 0.01), similarly as the coefficient of glucose assimilation (1.83 +/- 0.03 as compared with 1.25 +/- 0.15; p < 0.05). Indicators of the acid base balance did not differ. Recipients in group CP were however permanently treated with bicarbonate. CONCLUSIONS: With both transplantation method it is possible to achieve compensation of diabetes close to normal. The carbohydrate tolerance is however better after transplantation of the whole pancreas. PMID- 9340188 TI - [Patients with familial combined hyperlipidemia and smoking]. AB - BACKGROUND: Evaluation of the negative impact of active smoking on lipid and lipoprotein serum levels and the relationship with body mass index (BMI) and waist/hip ratio (WHR). METHODS AND RESULTS: The group was formed by 178 (77 men and 101 women), mean age 54 years (SD 6.2 attending the lipid out-patient department at the Third Medical Clinic, First Medical Faculty, Charles University Prague. Attention was paid to whether the patients were treated with hypolipidaemic agents or not. A statistically significant difference (p < 0.05) was found between smokers and non-smokers as regards total cholesterol (mean values and SD) in the group of 80 treated women (smokers 7.84 mmol/l, SD 1.21). Significantly higher values of LDL cholesterol in smokers as compared with non smokers (6.00 mmol/l SD 2.08 vs. 4.8 mmol/l, SD 1.26) were recorded in the group of treated women. Other results were also to the disadvantage of smokers (with the exception of non-treated women), though the difference was not statistically significant: non-treated female smokers had a higher LDL cholesterol (4.24 mmol/l, SD 1.42) as compared with non-treated non-smokers (5.3 mmol/1, SD 0.60). In treated men the LDL cholesterol value in smokers (5.20 mmol/l SD 1.20) were also higher than in non-smokers (4.54, SD, 1.15). Triacylglycerols in women (smokers vs. non-smokers): in the group of treated women 3.11 mmol/l, SD 4.86 vs 1.94 mmol/l SD 1.08, in the group of non-treated women 3.74 mmol/l, SD 4.77 vs. 1.94 mmol/l, SD 0.74. Triacyglycerols in men (smokers vs. non-smokers): in the treated men 3.87 mmol/l SD 3.54 vs. 2.62 mmol/l, SD 1.63, in the non-treated men 10.62 mmol/l, SD 9.86 vs. 2.86 mmol/l, SD 1.63. HDL-cholesterol in women (smokers vs. non-smokers): in the treated group 1.24 mmol/l SD 0.46 vs. 1.39 mmol/l, SD 0.35, in the group of non-treated men 1.54 mmol/l, SD 0.37, vs. 2.86 mmol/l, SD 1.64. HDL cholesterol in men (smokers vs. non-smokers): in the treated group 1.15 mmol/l, SD 0.30 vs. 1.27 mmol/l, SD 0.31, in the non-treated group 1.09 mmol/l, SD 0.40 vs. 1.15 mmol, SD 0.28. In female smokers treated and non treated and in non-treated male smokers the WHR values were higher, in treated smokers lower despite the surprisingly higher BMI. CONCLUSION: Active smoking has an adverse impact on serum lipid and lipoprotein levels in patients with familial combined hyperlipidaemia. PMID- 9340189 TI - [Trends in physical fitness in male white collar workers in the Czech population over the past 20 years]. AB - BACKGROUND: Common mortality and especially mortality on CAD is increased by low aerobic component of physical fitness. A study dealing with physical fitness changes of the Czech population has been still missing in our bibliography. The aim of the study was to evaluate physical fitness changes in the Czech male population since last 20 years. METHODS AND RESULTS: Physical fitness in 494 men in the age range of 20 - 60 years was compared to the men fitness examined 20 years ago. In the men of 20 - 29.9 years peak power (3.90 W vs 3.46 W, p < 0.01) and maximal ventilation (1.38 l.min-1 vs 1.24 l.min-1, p < 0.05) decreased. In the men 30 - 39.9 years old only peak aerobic power (37.2 ml.min-1 vs 35.7 ml.min 1, p < 0.05) slightly decreased. In the men 40 - 49.9 years old Vemax.kg-1 (1.38 l.min-1 vs 1.18 l.min-1, p < 0.001) and VO2max.kg-1 (35.6 ml.min-1 vs 32.4 ml.min 1, p < 0.001) severely decreased as well as in 50 - 59.9 years old men VEmax.kg-1 (1.23 l.min-1 vs 1.13 l.min-1, p < 0.05) and VO2max.kg-1 (32.7 ml.min-1 vs 29.4 ml.min-1, p < 0.001). CONCLUSIONS: Physical fitness of the Czech white-collar men decreased. The decline of physical fitness was related not only to lower exercise activity, but also to significant increment of body fat. Physical fitness, especially its aerobic component, decreased in Czech older men since last 20 years. PMID- 9340190 TI - [Magnesium balance in patients with spasmophilia. Relation to results of electromyography]. AB - BACKGROUND: The pathophysiological basis of spasmophilia is frequently magnesium deficiency and the therapeutic administration of magnesium salts has usually a favourable effect. However the parameters of magnesium balance are not always consistent with the results of electromyography. The objective of the present work was to test and interpret the relationship of results of these two basic diagnostic procedures indicated when spasmophilia is suspected. METHODS AND RESULTS: Thirty-three subjects (9 men and 24 women) with suspected spasmophilia were examined by non-invasive electromyography, using the technique of surface electrodes. All subjects had concurrently biochemical examinations: serum calcium and ionized calcium, serum magnesium (S-mg), magnesium in erythrocytes (ery-Mg) and magnesium in the blood haemolysate (H-Mg). In 29 patients and oral magnesium loading test was made with evaluation of the urinary Mg excretion after a constant Mg load (U-Mg). Statistical evaluation of the investigated parameters of the magnesium balance revealed a highly significant relationship between ery-Mg and U-Mg and H-Mg and ery-Mg (p < 0.005). A less close relationship was found between H-Mg and S-Mg (p < 0.05). Total and ionized calcium was in all examined subjects within the range of the arbitrary normal range. The EMG finding was positive (the finding of two and more multiplets in the ischaemic and hyperventilation test resp.) in 30 instances, i.e. in 91% of the examined subjects. In 72% there was agreement of the positivity of the EMG and magnesium deficiency (i.e. reduced values of ery-Mg and U-Mg), positivity of EMG combined with normal parameters of the Mg balance was recorded in 18%. In 6.1% of the examined subjects magnesium deficiency was confirmed combined with a normal EMG finding. CONCLUSIONS: Concurrent positivity of EMG and magnesium deficiency in 72% justifies the therapeutic administration of magnesium. In patients with a normal magnesium deficiency and positive EMG another cause of spasmophilia must be taken into consideration, incl. technical errors of interpretation of EMG results. A negative EMG associated with magnesium deficiency can suggest the central form of tetany, where magnesium treatment is also unequivocally indicated. PMID- 9340191 TI - [New drugs with positive effects on bones]. AB - The paper concerns the nontraditional treatment of osteoporosis using endogenous substances regulating bone metabolism, and also new drugs. NO in high concentrations decreases the activity of osteoclasts, scavenges superoxides which destroy connective tissue, and activates 1 alpha-hydroxylase in kidneys. Bone metabolism is effectively influenced by donors of NO or by modulators of NO synthase. Osteoclastic function is also inhibited by vitamin K. The administration of the vitamin is indicated in osteoporotic patients with proven vitamin K deficiency. Antiestrogens (tamoxifen), ipriflavon and analogues of wortmannin have antiresorptive activity. Under certain conditions parathyroid hormone (PTH) is anabolic for bone. The positive effect on bone was confirmed with the subcutaneous administration of small doses of PTH simulating physiologic pulsatile secretion, as well as the intact somatotropin-IGF-I (insulin like growth factor-I) axis. PTH is extremely useful, especially in osteoporosis induced by hypoestrinism. Somatotropin (GH) also has an anabolic effect on bone. The hormone stimulates bone metabolism with a prevalence of formation due to direct action on bone, as well as by means of IGF-I. Further growth factors with positive osteoprotic effect are TGF-beta (transforming growth factor-beta), FGF (fibroblast growth factor) and calcium conserving dihomogammalinoleic acid. Magnesium influences bone in different ways. It activates osteoblasts, increases bone mineralization, and enhances the sensitivity of target tissues (incl. bone) to PTH and 1,25(OH)2 vitamin D3, Under certain conditions however, magnesium can stimulate bone resorption. A more potent factor than magnesium is stroncium, which not only activates osteoblats but decreases the number of osteoclasts, thus abolishing bone resorption and enhancing formation. Bicarbonates are also favourable for bone. NaHCO3 together with potassium citrate stimulates osteoblasts and enhances bone mineralisation. In the review other prospective substances are also discussed. The osteoprotic effects of most of these factors were confirmed in vitro and in studies in animals, but their use in clinical practice is still a matter for investigation. Mutual interactions with classical osteoprotic drugs remain to be established. PMID- 9340192 TI - [Spontaneous remission of corticosteroid osteopenia after successful surgical treatment of Cushing's syndrome. A cross-sectional study]. AB - BACKGROUND: Osteoporosis is one of the most serious consequences of Cushing's syndrome. Only a small number of longitudinal observations on bone mineral density (BMD) in patients with treated Cushing's syndrome have been reported so far. To evaluate changes in bone mass in patients with Cushing's syndrome after surgical cure of the disease, BMD was evaluated cross-sectionally. METHODS AND RESULTS: BMD (DPX-L, Lunar) was measured in the lumbar spine and femoral neck (i.e. in skeletal areas with high proportion of trabecular and cortical bone, respectively) in 72 patients after successful surgical cure of Cushing's syndrome (8 men, 34 women before menopause and 30 women after menopause who were not on hormone replacement therapy). No other drug's interfering with skeletal metabolism were used. The reference group consisted of young healthy Czech women and/or men. The mean lumbar spine and femoral neck BMD increased to normal values within 3 and 5 years after surgery, respectively. In women after menopause, however, the significant positive relationship between BMD and time after surgery was negatively influenced by time after menopause. It is likely that several other factors contributed to the increase in bone mass in the patients (cure of hypercortisolism, recovery from hypogonadism and restoration of muscle strength). CONCLUSIONS: After surgical cure of Cushing's syndrome in premenopausal women and in men, BMD rapidly and substantially increases. The recovery is negatively affected by estrogen deficiency in postmenopause. PMID- 9340193 TI - [Effect of estrogens on cell adhesion molecules in thrombophilic states associated with high blood estrogen levels]. AB - BACKGROUND: Incidence of thromboembolism in pregnancy is relatively low regarding to many predisposing factors, inclusively the thrombophilia induced changes in coagulation. To evaluate this contradiction we presumed: 1. still not adequately evaluated role of the fibrinolytic system, 2. possible suppression of cytoadhesive molecules in bloodstream and consequently decreased activation of vascular endothelia by high levels of estrogens in the course of pregnancy. METHODS AND RESULTS: The tests of fibrinolysis (serum levels of fibrinogen, D dimers, tissue plasminogen activator (tPA) and plasminogen inhibitor 1 (PAI-1) and circulating cytoadhesive molecules (sE-selectin, sP-selectin and ICAM-1) were followed in a group of 66 pregnant women and compared with the unpaired Student's t-test with a group of 16 women after uncomplicated ovarectomy for benign disease. Results were compared with the serum 17 beta-estradiol levels. In pregnancy, significantly decreased levels of both selectins were found: sP selectin: 159 +/- 39 micrograms/l (SD) vs. 205 +/- 69 micrograms/l (p < 0.05), sE selectin: 33 +/- 12 micrograms/l (SD) vs. 43 +/- 17 micrograms/l (p < 0.05), meanwhile no significant difference in the concentration of ICAM-1 was found: 248 +/- 80 micrograms/l (SD) vs 285 +/- 101 micrograms/l (SD) (p > 0.05). In concordance, there were found significant negative correlations between serum levels of 17 beta-estradiol and sP selectin (r = -0.35, p < 0.01) and between serum levels of 17 beta-estradiol and sE-selectin (r = -0.21, p < 0.05) but not between 17 beta-estradiol and ICAM-1 (r = -0.1, p > 0.05). There were significant correlations between serum levels of 17 beta-estradiol fibrinogen (r = 0.46, p < 0.01), plasminogen activator I (PAI-1) (r = 0.38) and tissue plasminogen activator I (tPA) (r = 0.22, p < 0.05). No difference was found in serum levels of lipoprotein Lp(a) and naturally occurring inhibitor of the receptor for interleukin 1 (IL-Ira). CONCLUSIONS: This data supports the concept that the decreased serum levels of cytoadhesive molecules of sP-selectin and sE-selectin are dependent on serum estrogen levels and together with a new, estrogen induced, equilibrium of the fibrinolytic system suggest an explanation for the relatively low incidence of thromboembolic events in pregnancy. Decrease of cytoadhesive molecules may be one of explanations of favourable effects of estrogens on the development of atherosclerotic vascular changes. PMID- 9340194 TI - [Importance of determination of proliferation markers and hormone receptors in breast carcinoma]. AB - BACKGROUND: Cell cycle kinetic measures have been shown to have prognostic significance in breast cancer. The proliferative activity of tumors has been investigated with different approaches among which the use of monoclonal antibody MIB1 (against Ki-67 antigen) represents an easy and reliable means of assessing cell proliferation. The prognostic value of quantitative estrogen receptor (ER) and progesterone receptor (PR) immunohistochemical analysis has been also well established. This study was conducted in order to demonstrate our experience with an immunohistochemical detection of the markers of proliferation and hormone status in formalin-fixed paraffin-embedded samples of 1224 primary breast carcinomas. METHODS AND RESULTS: In this study, the proliferative activity of breast carcinomas was investigated using immunohistochemistry with the monoclonal antibody MIB1 (Immunotech). For a detection of ER and PR content in breast carcinomas, the antibodies ER1D5 (Dakopatts) and PR10A9 (Immunotech) were used. Immunohistochemical studies were performed on sections of formalin-fixed paraffin embedded tissues using the antigen retrieval method with a microwave oven irradiation. The MIB1 index was related to the histologic grade of ductal invasive carcinoma (p < 0.001). The study also revealed a significant correlation of the MIB1 index with ER values in grade II (p < 0.1) and grade III (p < 0.001), but not in grade I of ductal invasive carcinoma. Prognostic value of an immunohistochemical analysis of the MIB1 proliferative index, ER and PR status was determinated in this study. PMID- 9340195 TI - [The importance of calcium ions in the body]. AB - The review on the calcium ion is divided into two parts. One part contains so called basic physiological data on calcium: distribution on body fluids, tissues, its bond with proteins, daily turnover and calcium requirement in different stages of life, its absorption and excretion. In the second, special, part the author presents recent findings on calcium at a molecular and cellular level: calcium as the so-called primary messenger, types of intracellular communications, relationship with G-proteins, relationship of calcium with its regulators (vitamins D, parathormone, calmodulin). PMID- 9340196 TI - [Suggestibility, its origins and requirements]. PMID- 9340197 TI - A cluster of cytoplasmic histidine residues specifies pH dependence of the AE2 plasma membrane anion exchanger. PMID- 9340199 TI - [Digital radiology]. AB - Digital radiology is the integration of established and new digital imaging modalities into a system which enables the filmless communication of radiological information. It is becoming an indispensable tool in daily medical practice, and yields improved, faster medical communication, reduces the patient's dose and saves 307 307 archivation costs. Some knowledge of the inherent features of digital images and some open-mindedness for a change in viewing and workflow habits is necessary to make full profit of these possibilities. This paper describes digital radiography and the aspects of radiological networking based on 5 years experience with one of the very first filmless radiology departments worldwide. PMID- 9340198 TI - The role of nitric oxide in the neural control of nasal fluid production. AB - The production of nasal fluids serves an important role in the protection of the upper respiratory system, but can also be a troublesome symptom of rhinitis. The chief sources of nasal fluids are serous and mucous glandular secretion, epithelial goblet cell exocytosis, and exudation from submucosal blood vessels. This study was designed to investigate the role of nitric oxide in neurogenically mediated nasal vascular exudation and mucus secretion. A rat model of the naso nasal reflex was developed in which one nasal cavity was challenged with histamine while albumin and mucin production were measured in the continuously perfused contralateral side. Histamine challenge was associated with a significant rise in contralateral albumin and mucin content. Perfusion with a nitric oxide synthase inhibitor (L-NAME) in the nasal cavity contralateral to nasal challenge was found to block albumin leakage, but not mucin secretion, on that side. The inhibition of vascular exudation was overcome by the addition of L arginine, the natural substrate of nitric oxide synthase, to the perfusate. Treatment of the ipsilateral nasal of the ipsilateral nasal cavity with L-NAME did not significantly after the contralateral response. A high correlation was observed between albumin and mucin concentration in the perfusate. These findings indicate that NO is a mediator of the effector arm of the naso-nasal reflex that increases vascular permeability, but is not involved in the sensory nerve afferent pathway or in reflex mucin release. Further elucidation of the role of NO in nasal physiology may lead to novel pharmacotherapeutic approaches to the treatment of allergic and non-allergic rhinitis. PMID- 9340200 TI - [Value and importance of magnetic resonance tomography]. AB - After 14 years of clinical practice, magnetic resonance imaging (MRI) has gained an important place among the imaging methods. For a physician not permanently occupied with these problems, the differential indications of MRI compared to ultrasound and computertomography (CT) are difficult to understand. Knowledge of capabilities and limitations, benefits and drawbacks, contraindications, future trends, and, in an era of shortage of financial resources, knowledge of some essential economic aspects will be helpful. This publication tries to present these aspects in a compact form were singular problems of MRI diagnostics can only partly be dealt with. PMID- 9340201 TI - [The anxious patient during magnetic resonance tomography (MRI) examination. Health care economic aspects of patient education]. AB - Due to the increasing debate of limiting costs in the public health system, the use of magnetic resonance imaging (MRI) has been under discussion for several times. Diagnostic imaging by means of MRI is an expensive but informative and safe method. Instead of looking on cost-benefit analysis, it is shown how the efficiency of MRI can be increased by improving patient information about this imaging method. Because of the technical conditions, up to 40% of the patients show anxiety-related-reactions. These reactions include slight discomfort up to claustrophobic reactions and panic attacks. Therefore, correct and complete informing of patients prior to MRI examinations by the physicians is very important. The given information should include technical aspects about MRI and in addition, the physician has a chance to evaluate the patients' disposition to anxiety-related-reactions. To prepare risk patients for successful MRI technics like music, prone positioning, sedatives and relaxation exercises are available. By taking patients' anxiety into consideration and using the techniques mentioned above, unnecessary costs for unsuccessful or repeated MRI examinations can be avoided and the overall costs for clinical diagnostics will be reduced. PMID- 9340202 TI - [Interventional radiology--current status and future importance]. AB - The main methods of interventional radiology are described under clinical aspects. Treatment in peripheral arterial occlusive diseases by angioplasty and special other methods (stents, thrombectomy), as well as in bleeding management by embolization or different diagnostic and therapeutic techniques associated with oncologic diseases are performed. Minimal-invasive procedures are one of the most important courses of the practical medicine in the future. PMID- 9340203 TI - [Nuclear medicine diagnosis--principles and indications]. AB - Diagnostic procedures in nuclear medicine are functional studies of localized processes. The gathered informations are generally different to those acquired by imaging methods such as ultrasound and x-ray. Frequently, both methods are complementary. This review provides survey about the scintigraphic diagnostic procedures of following organs: Endocrine organs, especially the thyroid gland; heart, especially myocardial perfusion and viability, functional studies of the kidney, lung ventilation and perfusion, bone, bone marrow and inflammation, brain, gastrointestinal tract diagnostics and tumour scintigraphy. PMID- 9340204 TI - [Ultrasound diagnosis of rheumatic/inflammatory joint diseases]. AB - Ultrasonography is an inexpensive and readily available imaging technology for joints and surrounding soft tissues. Examination usually requires only a few minutes, is safe and can be repeated frequently because no radiation is involved. The method depends much on the training and skills of the examiner and sonographic pictures should always be interpreted in context with history and clinical findings. Therefore, the same physician, who does the physical examination, should also perform the ultrasound. Then, this method has the potential to serve as the 'rheumatologist's extended finger'. PMID- 9340205 TI - [High resolution 20 MHz ultrasound diagnosis in dermatology for noninvasive imaging of malignant melanomas]. AB - For years, high-resolution b-scan ultrasound is a well established technique in dermatology regarding the preoperative determination of thickness of skin tumors like malignant melanoma or basal cell carcinoma. Most tumors appear as echopoor areas in high-frequency sonography. Until today, a differentiation between skin tumors by means of high-frequency ultrasound is not possible. This is also true concerning the differentiation between the tumor tissue itself and its subtumoral inflammatory infiltration. Therefore, this lack of differentiation leads to the fact that sonometric thicknesses of tumors often exceed the originally measured tumor thickness in histology. By means of three-dimensional reconstruction of serial b-scan images, it is possible to determine each tumor's volume and surface as well as the topographical arrangement of echopoor areas and sonographical structures. Besides high-frequency ultrasound, additional non-invasive techniques like MRI, ultrasound- or laser-doppler, computer-aided image analysis and epiluminescence microscopy are also available for diagnosing malignant melanoma. Epiluminescence microscopy is used today as a standard technique in the diagnosis of malignant melanoma due to the possibility of differential diagnosis. Latest techniques like OCT (optical coherence tomography) are not ready yet to be applied regularly in diagnosis of tumors in the field of dermatology. PMID- 9340206 TI - [Ultrasound morphology of peripheral lymph nodes]. AB - Ultrasonography is the predominating method for the diagnosis of altered superficial lymph nodes. According to well defined criteria, the differentiation between benign and malignant nodal disease is possible with a high accuracy. The major concern of diagnosis is the characterization of enlarged lymph nodes, the staging and the long-time control of patients with tumors of the head and the neck and of patients with melanomas. Exemplary typical appearances of altered lymph nodes will be explained. Beside size and shape of the node, it is important to have a close look to its internal structure. The central echogenic hilus, the nodal cortex and their relation has to be noted and excentric hypoechoic structures must be described. According to the literature and to our own experience, it will be shown how reliable the commonly used diagnostic systems are. PMID- 9340207 TI - [Quality assurance in radiology]. AB - This review goes into important rules that users of radiology devices have to deal with. The legal requirements, the standards of the medical and common self administration are seen from the view of quality improvement, and the overlapping of the different areas are made clear. Big and important parts could not be taken into account in this review as they might be state or even community dependent. It is almost impossible for one physician to get access to all these informations. Therefore, it seems to make sense to offer a focused description which allows an overall view on the current status of norms. It is supposed to encourage further contributions to achieve reasonable and commonly accepted procedures in order to prevent mostly unobjective criticism against this important diagnostic tool. PMID- 9340208 TI - [Quality assurance in roentgen diagnosis. 1996 results and problems from the viewpoint of the Medical Section in accordance with ordinance 16 of the roentgen regulation of the Saxony Regional Medical Group]. AB - The "Arztliche Stelle gemass RoV" was established in Saxony in 1992. Structure and results of quality assurance in radiological diagnostics are described briefly. Quality checks delivered acceptable results for the majority of X-ray devices. Typical errors are shown and explained by using some X-ray pictures. PMID- 9340209 TI - [Therapy recommendations of the Drug Committee of the German Medical Association- an instrument for quality assurance in drug therapy. Drug Committee of the German Medical Association, Cologne]. AB - The growing flood of information on drug therapy, which is increasingly overwhelming the individual physician, and the simultaneous demand for high quality, individual therapy, call for guidelines recommending appropriate therapeutic measures for a given indication. The Drug Commission of the German Medical Profession, a scientific committee of the German Medical Association, has set itself the task of providing assistance in this context by elaborating scientifically-based, practice-oriented therapeutic guidelines. These guidelines were drawn up in accordance with a defined procedure and represent the consensus reached between the appropriate independent experts, the general practitioners and the members of the executive committee of the Drug Commission. This review presents the background, aims, procedures and criteria for assessment involved in the elaboration of the guidelines and attempts to define the position of the therapeutic guidelines in comparison to similar activities of other institutions in Germany. PMID- 9340210 TI - [Practice guideline--peripheral arterial occlusive disease. Drug Committee of the German Medical Association]. PMID- 9340211 TI - [Incidence and outcome of pregnancies with contraception]. AB - In 1992, 1531 women aging 25-45 years from 5 urban and rural regions were interviewed about infertility and subfecundity within the German part of a study by the European community. 1248 of them had a positive reproductive history with 3018 pregnancies. 400 (= 13.25%) occurred during contraception. This affected 296 women (= 19.6% of all women). Smoking seems to have a promotion effect. The risk of ectopic pregnancies seems to be higher in contraceptive users. The highest fetal loss was found in women using an IUD. There were no differences in the course and outcome of pregnancy between users and nonusers of contraceptives. About 40% of the women in the corresponding group reported an irregular application of the contraceptive technique. PMID- 9340212 TI - [A new development in quality management: first complete European Foundation for Quality Management assessment of a German Clinic]. PMID- 9340213 TI - [Comparison of proliferative activity in malignant lymphomas determined by immunohistologic methods with various antibodies]. AB - Proliferation Index (PI) was evaluated in a group of 48 malignant lymphomas. Comparison of 3 antibodies showed that DAKO-EPOS Anti-PCNA/HRP and MIB-1 (Immunotech) suite to routine PI evaluation. Low figures of PI were obtained when using DAKO-EPOS Anti-Ki-67/HRP which did not even discriminate between prognostic groups of malignant lymphomas. PMID- 9340214 TI - [Clinicopathologic correlation of interstitial pulmonary processes]. AB - "Not-specified interstitial" lesions represented mostly cryptogenic fibrous alveolitis, extrinsic allergic bronchioloalveolitis prevailed among "disseminated" lesions, "infiltrative" lesions were proved to be extrinsic alveolitis or disseminated carcinoma. Among more definitely formulated suppositions of the clinician, cryptogenic alveolitis was proved in more then 50% of cases often modified by extensive fibrosis, organized pneumonia, cholesterol pneumonia or minute desquamative pattern. A similar relation of diagnostic correctness was found in allergic bronchioloalveolitis suprisingly often modified again by fibrosis and cholesterol pneumonia. Valid diagnoses were found in sarcoidosis, presumed Goodpasture syndrome harboured only pneumoconiotic stigmatization. In brief: Bioptical diagnosis nearly always replaced a vague clinical evaluation whereas definite nosological formulations were proved, turned more exact or changed in equal proportions, biopsy was not found contributory but in two cases. PMID- 9340215 TI - [Alveolar adenoma of the lung (case report)]. AB - A case of a rare benign solitary pulmonary neoplasm of alveolar adenoma type in a 42-year-old man was presented. The tumor consisted of a glandular component which reacted with antibodies against cytokeratins CAM5,2, AE1-3 as well as against epithelial membrane antigen (EMA) in superficial parts of cytoplasmatic membrane, while actin and desmin were positive in the stromal component. Ultrastructurally, glandular component represented chiefly type II pneumocytes, while pneumocytes of type I occurred rarely. Stroma of the tumour consisted of myofibroblasts, fibrocytes, and collagen fibrils. PMID- 9340216 TI - [Histiocytic reaction in the lymph nodes after total hip joint endoprosthesis surgery]. AB - In two patients with hip replacement, the pelvic lymph nodes were resected during prostatectomy for carcinoma. The striking histiocytosis was present in lymph nodes. The cytoplasm of histiocytes was bulky, granular, PAS positive, with KP1, lysozyme and alfa-1-antitrypsin positivity. In polarized light microscopy, the birefringent, needle-like particles corresponding to polyethylene or polyester respectively were seen. Those changes are characteristic for reactive lymphadenopathy in regional lymph nodes following joint endoprosthesis. PMID- 9340217 TI - [Transmyocardial laser revascularization--histopathologic findings]. AB - Findings in a 56-year old man who died 3 months after laser heart surgery were described. In the site of surgery (anterior and lateral wall of left heart ventricle) patent slit-like channels we found that may or may not have been performed by laser. Channels were indistinguishable from lymphatics. In second case, a 59-year old woman died 5-days after laser revascularization. Channels filled by fibrin, cellular debris and polymorphonuclear leukocytes were found, none of them patent. PMID- 9340218 TI - [Problem-oriented training in pathology--first 5 years' experience]. AB - The article explains some of the main characteristics of problem-based learning (PBL). Five-year experience with application of PBL in pathology is described. Student appreciation of PBL is complemented by assessment in the literature. PMID- 9340219 TI - [Tests for teaching pathologic anatomy]. AB - A teaching database for pathology was prepared comprising around 1250 questions. Five answers belonged to each question, only one of them being correct. Preparation of questions was partly based on testing sets from Medical School of the University of Loma Linda, Ca., partly on those used in 2nd Department of Pathology, Masaryk University Medical School, Brono. In addition, set of computer programmes for automatic generation, printing and evaluation of tests was prepared. They were verified with Brno students. PMID- 9340220 TI - [Prognosis in breast carcinoma]. PMID- 9340221 TI - ["Crazy women deceive the imprudent man": recognition and defamation of female and male physicians, 1450 to 1700]. PMID- 9340222 TI - [Leprosy in a mural of Monreale]. PMID- 9340223 TI - [Reconstruction of the distribution of wound surgeons in the Wurttemberg kingdom. A contribution to the social history of routine surgeons in the 19th century]. PMID- 9340224 TI - [The destiny of the Latvian physician Lasar Kopelowitsch (1902-1941) and psychobiology]. PMID- 9340225 TI - [Temporary legal protection in fee conflicts. When can the social court help]. PMID- 9340226 TI - [Hygiene plans for ambulatory surgery practices]. PMID- 9340227 TI - [Surgery on risk patients in emergency and elective situations I]. PMID- 9340228 TI - [Risk assessment]. AB - The aim of preoperative risk analysis is to reduce postoperative morbidity and mortality by identification of compromised organ function which can be improved by targeted preoperative measures and problem-oriented postoperative therapy. Ideally, therefore, preoperative risk assessment, influences the timing of the surgical procedure, the choice of the surgical approach, and the postoperative management. Identification of preexisting relevant disorders that may influence the postoperative course is an essential prerequisite of risk analysis. While preoperative risk analysis today gains in importance with the increasing extent of elective surgical procedures, preoperative risk evaluation still plays a minor role in the emergency situation when the given patient-dependent risk usually has to be accepted. Objective risk evaluation depends on the general and nutritional status, the pulmonary, cardio-vascular, hepatic, and renal function, and the cooperation of the patient. These factors, however, clearly have to be seen in relation to the type and extent of the planned surgical procedure. The selection of additional tests of organ function, exceeding the standard tests required prior to any surgical intervention, must be guided by the extent of the planned surgical procedure, the physiologic alterations associated with the surgical procedure, and the suspected underlying organ dysfunction. Generally accepted multi-factorial classification systems to identify patients at risk for a wide spectrum of surgical procedures are currently not available. Using the model of esophagectomy in patients with esophageal cancer we could, however, demonstrate that a quantitative assessment of the peri- and postoperative risk based on preoperatively available physiologic parameters is possible and markedly reduces postoperative mortality when applied prospectively. The development and validation of similar risk-score systems for other surgical procedures should be considered. PMID- 9340229 TI - [The patient with respiratory problems]. AB - Despite improvements in operative and anesthesiological techniques, respiratory problems in surgical patients have been minimized but not eliminated. In addition to risks which are typical for the individual patient, the perioperative respiratory morbidity is affected by anesthesiological manipulations as well as the operation and the nature of the operation (elective versus emergency). In this paper, after describing anesthesia-associated disturbances of the respiratory situation together with worsening due to the disease in patients with COLD, techniques and methods for therapy, prophylaxis, and prognostic assessment are delineated. Two examples are given for patients with respiratory problems (abdomino-thoracic esophageal resection as a example of local trauma in patients with numerous preoperative risk factors and acute necrotizing pancreatitis to describe the sequelae of a toxic process). The essence of our discussion is that, prognostically, preoperative diagnosis is of reduced value. Only a synopsis of clinical findings together with spirometry and blood gas analysis appears to be relevant. Early mobilization in conjunction with excellent postoperative pain therapy is of utmost importance, which is equivalent to the almost routine placement of a patient controllable epidural analgesia technique. These concepts have shown in the two patient groups described that respiratory morbidity may be reduced significantly. Cooperation between surgeons and anesthesiologists, which is characterized by complete and mutually high competence on both sides, is essential for successfully managing patients at increased respiratory risk. PMID- 9340230 TI - [Surgery in patients treated with anticoagulation during emergency and elective interventions]. AB - Patients under oral anticoagulation with coumarin derivatives have a variable perioperative thromboembolic risk which necessitates continuation of their thromboembolic prophylaxis during elective and emergency surgery. Because of their better handling unfractionated heparin and low-molecular-weight heparins are used most often for this purpose. The overlapping effect of coumarin and heparin therapy requires a close daily monitoring of the clotting inhibition (partial thromboplastin time, thromboplastin time, thrombin time). In elective surgery coumarin therapy is interrupted 2-3 days preoperatively; in emergency cases vitamin K or fresh frozen plasma have to be substituted. Patients under heparin therapy or prophylaxis are easier to handle, because the effect of heparin disappears in a few hours after stopping the treatment. In the immediate postoperative phase the heparin application is interrupted for 6 h because of increased bleeding risk. It is important to take into account additional risk factors like the underlying disease, disturbances of platelet function, liver diseases and renal insufficiency. PMID- 9340231 TI - [Surgery in immunosuppressed patients with emergency or elective indications]. AB - Immunosuppressive therapy and its influence on perioperative pathophysiology present special challenges in the event of surgical intervention. Immunosuppressive agents alter the patient's response to surgical stress and infectious complications. The often masked signs, even in the case of severe infection, require a high index of suspicion to establish the diagnosis. This may result in a fatal delay of therapy. In addition, the immunosuppressed state increases the patient's susceptibility to infection and leads to an impairment of wound healing. Therefore, careful perioperative clinical monitoring of the patient and complete control of the immunosuppressive therapy are mandatory. Elective operations in immunosuppressed patients should be performed with special caution regarding the potential perioperative risks for the patient and the graft. On the other hand, if there is evidence of, for example, an acute abdominal event, a more aggressive approach is required to rapidly establish the diagnosis and institute appropriate therapy. From the surgical point of view, special emphasis should be placed on wound closure and on anastomotic sutures when operating on a patient receiving immunosuppressive therapy. PMID- 9340232 TI - [Value of preoperative blood coagulation analysis for assessment of hemorrhage risk in general surgery]. AB - Coagulation studies, i.e. platelet count, prothrombin time (PT) and activated partial thrombin time (aPTT) are commonly employed preoperatively to identify patients at risk. In a retrospective study we evaluated the usefulness of these screening tests to predict postoperative bleeding in 1447 patients with abdominal and thoracic surgery. Forty-six patients (3.2%) experienced postsurgical bleeding. 12.2% of our patients had abnormal coagulation studies. The sensitivity of abnormal coagulation studies with respect to postoperative bleeding was 23.9%. The sensitivity of the parameter "patient at risk", i.e. patients with suspected coagulopathies due to drugs or disease of the liver or kidney, was 56.5%. Thirty four out of 1008 patients without risk factors had abnormal coagulation tests but an uneventful postoperative course. Preoperative screening of PT and aPTT should be reserved for patients with known or suspected inherited or acquired coagulopathies. PMID- 9340233 TI - [Results of deep rectum resection and intersphincteric rectum excision]. AB - Low resection and intersphincteric extirpation of rectal cancer in the distal third of the rectum has become an accepted sphincter-saving method. From December 1990 to December 1994, 42 patients (17 women and 25 men) with a mean age of 67.2 years had a low resection or extirpation of the rectum at our institution. Eighteen patients received a transanal sutured anastomosis, 24 a stapler anastomosis. We had a lethality rate of 2.5% and a anastomotic insufficiency rate of 14%. PMID- 9340234 TI - [Late results of Childs-Phillips mesenteric plication for therapy and prevention of small intestine ileus]. AB - During the past 7 years 45 patients have been operated upon using the Childs Phillips method. Of those, 37 were subsequently examined for the study--7 patients had died in the meantime. None of the deaths occurred as a direct result of transmesenteric small-bowel plication. An early recurrence of intestinal obstruction occurred in 4.4% and a laparotomy was repeated. During the most recent examinations 86.5% of those patients checked had (virtually) no complaints -91.9% based upon the Visick classification. A subtotal intestinal obstruction occurred during the period of the study in 8.1% of cases, but could be conservatively treated. Up until the most recent examination there were still no instances of a late recurrence. Most intestinal obstruction recurrences are due to errors specific to the technique and are early recurrences. On the basis of our results, we are of the opinion that plication in the presence of existing peritonitis, as well as partial plication, is acceptable. PMID- 9340235 TI - [Effectiveness of preoperative diagnosis in scintigraphically cold thyroid nodule]. AB - PURPOSE: To compare the diagnostic efficacy of different imaging procedures carried out in patients with scintigraphically cold thyroid nodules. METHODS: The preoperative imaging procedures carried out in all patients operated on for cold thyroid lesions at the Department of Surgery of the University of Cologne were recorded in prospective manner for 1 year. Special attention was paid to the difference between in-patient and out-patient management. Diagnostic accuracy was assessed by comparison with the intraoperative findings and histology. RESULTS: The combination of ultrasonography and fine needle aspiration cytology proved effective. Computed tomography should be reserved for the preoperative assessment of large thyroid tumors. Repeat examinations seldom gave additional information. CONCLUSION: Standardized preoperative investigations may be performed on an out patient basis, provided attention is paid to quality assurance. PMID- 9340236 TI - [Hormone inactive neuroendocrine tumors of the pancreas]. AB - During the past 10 years (1987-1996), 842 laparotomies were performed for pancreatic or periampullary neoplasms; in 25 patients (2.9%) a neuroendocrine tumor was diagnosed. In 19 of these 25 patients (76%) a non-functioning endocrine tumor and in 6 patients (24%) a hormone-active tumor (four insulinoma, one gastrinoma, one VIPoma) was found. Of 19 non-functioning neuroendocrine tumors, 14 were malignant. The resection rate of these malignant tumors was 78.6% (11 of 14; 3 resections were palliative); in comparison, the resection rate of ductal pancreatic carcinoma in our hospital was 28.1%. The probability of 5-year survival amounts to 73% after surgical resection in malignant endocrine tumors and to 19% in ductal pancreatic carcinoma (including palliative resections). As it is not always clear whether non-functioning endocrine tumors are benign or malignant, oncological resection is recommended. Adjuvant chemotherapy seems not to be necessary. PMID- 9340237 TI - [Cronkhite-Canada syndrome. Epidemiology, symptoms, morphology and therapy based on a case report and literature review]. AB - Juvenile polyposis was first described by Cronkhite and Canada in 1955. This disease is characterized by juvenile intestinal polyps and ectodermal abnormalities. The etiology of Cronkhite-Canada syndrome (CCS), however, is still not well understood. Interestingly among patients with CCS a significant correlation (16.5%) with intestinal carcinomas has been observed. Thus, malignant transformation and/or genetic predisposition may be involved in the initiation of the disease. In the following, epidemiology, symptoms, morphology and therapy of CCS are discussed. Our examinations are based on studies reported in the literature and on a case report of a female patient who developed a colon carcinoma 2 years after initial diagnosis of CCS. PMID- 9340238 TI - [Does a shower put postoperative wound healing at risk?]. AB - A prospective randomized study was designed to determine the effect of postoperative water contact on tissue healing. A total of 121 patients undergoing open hernia repair was divided into two groups. The first group was permitted to shower postoperatively, allowing the wound to come into direct contact with the water, the second group was instructed to keep the incision dry. Water contact, wound healing and patient satisfaction were assessed. There was no difference in wound healing between the two groups and no manifest infection. PMID- 9340239 TI - [First experiences with unreamed AO intramedullary nail in treatment of femoral shaft fractures]. AB - The unreamed femoral nail (UFN) system, with its numerous proximal interlocking options, allows a minimal invasive surgical procedure for the treatment of nearly all femoral fracture patterns. Sixty-six fractures, 5 cases of osteolysis or pathologic fractures, 2 limb shortenings and 1 lengthening (monorail technique) and 3 cases of pseudarthrosis were stabilised with the UFN from July 1994 to December 1996. The fractures were analysed according to the AO classification. We found 31 polytrauma patients with an mean ISS of 21.8 and a mean PTS of 25.4. Most of the multiply injured patients (n = 26) were stabilised with the UFN primarily. Follow-up of 44 patients ranged from 4 to 18 months postoperatively. According to our clinical and radiological score the results were excellent in 34% of cases, good in 36.3%, poor in 20.4% and bad in 9%. Average fracture healing time was about 9.8 weeks. PMID- 9340241 TI - [Primitive neuroectodermal tumor]. AB - The primitive neuroectodermal tumor is a rare soft tissue neoplasm occurring in children and young adults. It derives from a carcinogeneic alteration of pluripotent neural crest cells, caused by a balanced reciprocal translocation t(11;22) (q24;q12). Treatment of this undifferentiated, extremely malignant small cell tumor is carried out in compliance with the soft tissue trail (CWS) from the German Society of Pediatric Oncology. Biopsy-proven diagnosis is followed by primary chemotherapy, which in 95% of cases leads to remission, allowing excision of the remainder of the tumor without mutilation and avoidance of intraoperative tumor cell dissemination. After excision, irradiation of the tumor site and two further sequences of chemotherapy are performed. If PNETs of the paravertebral region cause symptoms of paralysis and immediate surgery is required, postoperative chemotherapy, a second-look operation and irradiation are mandatory. Between 1986 and 1994, in cooperation with our pediatric and radiotherapy colleagues, we treated ten patients. In four patients (median age, 14 years) the PNET originated from the chest wall, in six patients from the paravertebral and retroperitoneal region. Five patients died after 20 months on average, while the remaining five patients are in full remission after 31, 46, 50, 51 and 91 months, respectively. PMID- 9340240 TI - [Intestinal schistosomiasis, a facultative precancerous condition? Review of the literature with reference to Schistosoma japonicum associated rectum carcinoma]. AB - After a latency period of 20 years, in a 39-year-old Austrian citizen of Chinese origin, a surgically removed rectal carcinoma, as well as the neighboring chronic inflammatory rectal mucosa with various degrees of dysplasia and one positive neighboring lymph node, showed helminthiasis in the histopathological examination, convincing us of a link between carcinoma and chronic helminthiasis. Whereas the etiological context between chronic infection by Schistosoma haematobium and endemic frequent urinary bladder carcinoma is considered a matter of fact, whether of not the incidence of intestinal carcinoma is increased in connection with chronic intestinal schistosomiasis is controversial. The etiological and pathogenetic link between helminthiasis and carcinoma should be considered in the same way as for other related inflammatory large-bowel diseases. In the sequence chronic inflammation-severe dysplasia, the formation of carcinoma could possibly occur. Besides a survey of trematodes parasitology and pathology, the link between rectal carcinoma and Schistosomiasis japonicum is pointed out by means of appropriate literature investigations. PMID- 9340242 TI - [Post-traumatic anterior interosseous nerve syndrome after supracondylar humerus fracture in a child]. AB - The interosseus anterior syndrome is a rare nerve compression syndrome that concerns only the motor branch of the median nerve. It is evidenced by paralysis of the M. flexor pollicis longus and M. flexor digitorum profundus II with weakness of flexion of the terminal joint of the thumb and index finger but without sensory loss. From the surgical point of view this complication has special importance because of pediatric fractures of the distal humerus. In this paper we discuss the differential diagnosis, therapy and prognosis of this nerve injury by presenting an illustrative case of a 7-year-old boy suffering a supracondylar fracture of the humerus with a post-traumatic anterior interosseus nerve syndrome. PMID- 9340243 TI - [The Vollmer hook--a new abdominal retractor]. AB - Here, we present the so-called "Vollmer retractor", which can be fixed on to the operating table. It allows an optimal transference of the traction force on to the border of the wound, both in upper and lower abdominal interventions. In this way, as shown by our recent experience, we gain an optimal view of the subdiaphragmatic area and into the small pelvis. PMID- 9340244 TI - [Percutaneous ultrasound controlled drainage of large splenic abscesses]. AB - Because of the rare incidence of splenic abscesses and bleeding, only a few cases are described in the literature on percutaneous drainage of splenic abscesses. We report on eight cases (three male, five female, average age 74.1 +/- 10.76 years) of percutaneous, sonographically controlled catheter drainage of large splenic abscesses. RESULTS: Percutaneous, sonographically controlled drainage of splenic abscesses was feasible in all eight cases in the last 5 years (trocar technique, drain: 12-16 F, abscess contents 70-750 ml). In seven of eight cases, therapy with percutaneous, sonographically controlled drainage of the splenic abscess and rinsing of the abscess cavity over several days was successful. In two cases, however, a recurrent abscess had to be drained repeatedly with sonographic control, and this was successful. In one case a colitis-related fistula prevented successful drainage of an infected subcapsular splenic hematoma. The following splenectomy, however, proved the infected hematoma to be completely drained. In this study there were no complications like bleeding, injury of pleura or colon. The advantages of percutaneous drainage are: the spleen is not removed; conservative therapy is beneficial, particularly in multimorbid patients with a high surgical risk; there is no transmission of bacteria; the method is safe and effective. PMID- 9340245 TI - [Combined pancreas-/kidney transplantation as a standard procedure in therapy of kidney failure in type I diabetic patients]. PMID- 9340246 TI - [Bile duct drainage after laparoscopy]. PMID- 9340247 TI - [Clamping the tip of the appendix in appendectomy]. PMID- 9340248 TI - [Macrophages as production sites of HIV]. PMID- 9340250 TI - [Lymphomatoid granulomatosis--remission induction with interferon-alpha 2b]. AB - HISTORY AND ADMISSION FINDINGS: A 72-year-old woman was admitted to hospital because of nonproductive cough, acrodistal sensorimotor axonal polyneuropathy, fatigue and 10 kg weight loss over the preceding 9 months. INVESTIGATIONS: Chest radiogram showed multiple round foci in both lungs. No organs other than the lungs and the peripheral nervous system were affected. Open lung biopsy revealed a pleomorphic angioinvasive lymphocytic infiltrations. DIAGNOSIS, TREATMENT AND COURSE: The findings indicated lymphomatoid granulomatosis (LG). Wegener granulomatosis was excluded on the basis of the clinical, serological and histopathological findings. Treatment with alpha-interferon (IFN-alpha) produced complete remission within 4 months. Interruption of treatment immediately resulted in a recurrence. CONCLUSION: Together with previously reported cases this patient's response to IFN-alpha suggests that IFN-alpha is a less toxic alternative to cyclophosphamide and corticosteroid administration in the early stages of LG. PMID- 9340249 TI - [The Creutzfeld-Jakob disease. A sphinx of current neurobiology]. AB - BACKGROUND: Prospective epidemiological studies are being employed to determine the incidence and possible risk factors of Creutzfeldt-Jakob disease (CJD) in five European countries in which bovine spongiform encephalopathy (BSE) occurs at different rates of incidence. PATIENTS AND METHODS: Using a voluntary reporting system throughout the Federal Republic of Germany, suspected cases of CJD were investigated and the incidence calculated. Possible risk factors in patients and control groups were obtained by questionnaire. Serum and cerebrospinal fluid samples served to delineate genetic forms and distinguish the disease from other major dementias. RESULTS: A total of 544 patients with suspected CJD, reported in Germany between 1993 and 1997, were examined. 232 (plus 27 investigated only neuropathologically) were confirmed as definite or probable, an annual incidence per million population of between 0.76 (for 1994) and 0.98 (for 1995), similar to figures from other European countries. In Great Britain, the cases of "new variant" CJD, not yet observed in Germany, were excluded from the calculation of incidence. So far, dementia in the family and handling of horn shavings have been identified as risk factors. A rise in the concentrations of neurone-specific enolase and of S100 protein as well as the demonstration of certain proteins in cerebrospinal fluid (p130/ 131 and 14-3-3, respectively) have been shown as being diagnostically superior to EEG changes. INTERPRETATION: There has so far been no increase in the incidence of CJD within Europe. However, the occurrence of the new variant in Great Britain requires long-term monitoring. The diagnostic criteria used for this can be improved by biochemical methods. PMID- 9340251 TI - [Photochemotherapy with indocyanine green in cutaneous metastases of rectal carcinoma]. AB - HISTORY AND ADMISSION FINDINGS: Six weeks before admission of a 47-year-old man with known rectal carcinoma, small nodular metastases had occurred over the front of both his thighs. Examination showed many aggregatet cuti-color or livid nodes, diameter 0.5 cm. The patient's poor general condition excluded the usual palliative measures. TREATMENT AND COURSE: Photochemotherapy of the skin metastases with indocyanine green (ICG; absorption maximum 805 nm), a non-toxic dye approved for diagnostic purposes, was undertaken on a trial basis. The dye, being bound to plasma proteins, is retained in the intravacular space. Immediately after administration of the dye (2.5 mg/kg intravenously) the skin metastases were irradiated by diode laser (lambda = 805 nm, 100 J/cm2, 3 W/cm2, radiation diameter 2 cm). This necrosed the metastases and clinically as well as histologically resulted in their complete disappearance with scarring of the treated area. CONCLUSION: This case illustrates the effectiveness of photochemotherapy with ICG against solid skin tumours of increased microvascular density. PMID- 9340252 TI - [Place of trans-myocardial laser revascularization in treatment-resistant coronary heart disease]. PMID- 9340253 TI - [Gastrointestinal complications in stem cell transplantation]. PMID- 9340254 TI - [Hospital administration's refusal to perform surgery due to previous exceeding of budget. Decision of the Gelsenkirchen Labor Court 20 December 1996]. PMID- 9340255 TI - [Unclear causes of death]. PMID- 9340256 TI - [Significance of the HbA1c values in type I diabetes mellitus]. PMID- 9340257 TI - [Pneumococcal vaccination following transplantation]. PMID- 9340258 TI - [Application of peroxyacetic acid as disinfection of large stalls using thermonebulization (a case report)]. AB - The method of application, the material properties of the disinfectant as well as the spacious circumstances decide about the effect of aerogeneous disinfection processes. Microbiological results and practical experiences clearly show that formaldehyde can be replaced by special formulations of peroxyacetic acid (PES) in the procedure of thermonebulization; on condition that a suitable technique is used. Perhaps the decay of agents but also defects in the distribution were results of a too high temperature of application. Characteristic foultry effects on the floor could be replaced by the help of Pulse-Jet-Instruments with a fog development below 80 degrees C. Beware of vaporizing PES-preparations exclusively by heat supply. PMID- 9340259 TI - [Effects of peptide YY on functions of the gastrointestinal tract]. AB - The present report gives a review about the localization, release and gastrointestinal actions of peptide YY in different animal species and in humans. Possible mechanisms of action, the physiological and pathophysiological significance of peptide YY and the role of peptide YY 3-36 are discussed. Finally, unanswered questions are specified. PMID- 9340260 TI - [Ultrasonography of the inner stifle joint in dogs with rupture of the cruciate ligaments]. AB - This paper gives an overview of the sonographic possibilities to examine the canine stifle joint with rupture of the cruciate ligament. Sonographic examination was performed in 100 surgical patients because of a lesion of the joint. Ultrasound sections are presented with corresponding sonographical findings and pathological changes. For stifle joint, a linear transducer with 7.5 Mhz proved to be useful. Both the lateral and medial meniscus could be inspected, as well as injuries of the menisci. As an indirect sign to a lesion, effusions of the joint were demonstrable quite sensitively. The cruciate ligaments could not be investigated correctly with ultrasound. Ultrasonography is used to investigate the soft tissue structures as a completion of the radiological examination. The scanning diagnostic technique is practicable with a low technical expenditure, it is not stressful for the patient and can be repeated for therapy controls. Disadvantageous is that ultrasound is prone to artefacts, caused by the thinness of the structures examined. PMID- 9340261 TI - [Case report: decreased laying performance as a contributing problem]. AB - In 2 laying hen flocks (housed in aviary systems) depressed performance, increased mortality and cannibalism were observed. Specific infectious diseases were excluded. The hygienic quality of the diet, however, based on self produced feed ingredients, was impaired. High concentrations of bacteria, moulds, and yeasts were found in the complete diet and also in the ingredients. These problems were eliminated by providing the birds with good quality feed ingredients. In a diagnostic feeding trial with a change in housing conditions hens showed a good performance. The poor feed quality, as demonstrated in this report, is regarded to be one of several factors, which have contributed to the problems observed in these flocks. Other negative environmental factors such as housing have to be eliminated. PMID- 9340262 TI - [Quality management in the serodiagnosis of European swine fever: results of comparative tests]. AB - In the course of inter-laboratory quality management comparative serological tests on classical swine fever (CSF) are conducted once per year. Results from tests carried out in 1994 and 1995 indicate that most regional diagnostic laboratories were able to classify the test sera correctly as CSF-positive, bovine viral diarrhea (BVD)-positive and negative, respectively. Difficulties were encountered in the differential diagnosis of CSF and BVD in neutralisation tests. There is a need to improve the standardization of CSF serology on the basis of a well established method in order to ensure reliability of test results and to enable comparison of results obtained from different laboratories. PMID- 9340263 TI - [Comparison of different BVD virus strains for their use in the differential diagnosis of classical swine fever--an attempt to standardize neutralization tests]. AB - Six bovine virus diarrhoea (BVD) virus strains were tested in the neutralization test for their use in the differential diagnosis in classical swine fever (CSF) serology. The aim of the investigation was to find a suitable BVD virus strain guaranteeing a safe differentiation of CSF- and BVD virus induced antibodies using permanent cell cultures (PK-15, MDBK). For test purposes the neutralizing antibody titres of 73 defined test sera were titrated against the CSF virus strain Alfort/187 as well as the BVD virus strains Grub, Paplitz, NADL, 1138/69, Stendal, 10421/Han 94). Tests were repeated fivefold. The level of mean antibody titres, the differences in titre to the homologous pestivirus strain, the standard deviation and the variation coefficient served as test criteria. The BVD virus strains Grub and NADL yielded the best results. In view of harmonization and standardization the BVD virus strain NADL in connection with a standard protocol for neutralization tests is recommended for the differential diagnosis in CSF serology. Due to the adaptation of the permanent cell-lines PK-15 and MDBK to horse serum a further source of contamination with non cytopathogenic BVD viruses under routine conditions can be excluded. PMID- 9340264 TI - [Hemoglobin derivatives in blood of cattle during during winter housing. Effect of age as well as developmental and functional conditions]. AB - Peripheral venous blood samples taken from pregnant and lactating dairy cattle, from young cattle and from new born calves as well were analysed for hemoglobin content, hematocrit, oxygen saturation, oxygen capacity, oxygen content and for the hemoglobin derivatives O2Hb, HHb, COHb, MetHb and SHb. Significant differences of mean values could be pointed out between age groups and breeds. Strong negative correlations exist between MetHb fraction and hemoglobin content in dairy cattle and in new born calves but not in young cattle. Elevated COHb and MetHb fractions were discussed in connection with erythropoesis, regeneration of erythrocyte mass and vascular adaptation. PMID- 9340265 TI - [Reducing weight with leptin]. PMID- 9340266 TI - [Angiotensin II antagonists]. PMID- 9340268 TI - [Familial combined hyperlipidemia; solution on the horizon?]. PMID- 9340267 TI - [What is known about the etiology and background of schizophrenia?]. PMID- 9340269 TI - [Nasal ventilation in treatment of acute respiratory insufficiency]. PMID- 9340270 TI - [Cryopreservation of sperm in cancer treatment-induced infertility]. PMID- 9340271 TI - [Arteriobiliary fistula as a complication of biliary surgery]. PMID- 9340272 TI - [Reflex dystrophy]. PMID- 9340273 TI - [Somatization as a clinical problem]. PMID- 9340274 TI - [Comments on articles of rheumatoid arthritis]. PMID- 9340275 TI - [Would not serppi be o.k.?]. PMID- 9340276 TI - [Per oral glucocorticoid is infective for viper bite]. PMID- 9340278 TI - [What is good research?]. PMID- 9340277 TI - [Can inhaled corticosteroid used for asthma cure allergic rhinitis?]. PMID- 9340279 TI - [Impact, quality or meditation?]. PMID- 9340280 TI - [Research physician education has started]. PMID- 9340281 TI - [EVO-funds are a shot for Finnish research]. PMID- 9340283 TI - [Research plan and follow-up of the project]. PMID- 9340282 TI - [Am I to be a researcher?]. PMID- 9340284 TI - [Medical databases, index books and library services]. PMID- 9340286 TI - [Scientific misconduct]. PMID- 9340285 TI - [The principles, planning and realization of a clinical trial]. PMID- 9340287 TI - [How to give a lecture]. PMID- 9340288 TI - [How to write a scientific paper]. PMID- 9340289 TI - [Is it worthwhile to utilize commercially research results]. PMID- 9340290 TI - [International research programs and training of researchers]. PMID- 9340291 TI - [The new biology in medical education--an essential but not a sufficient paradigm]. PMID- 9340292 TI - [Scientific research of experiences: history of ideologies and today]. PMID- 9340293 TI - [Where to get funds for research--the possibilities for getting funds as a young researcher]. PMID- 9340295 TI - The James Young Simpson Legacy, 4-6 September 1997. Abstracts. PMID- 9340294 TI - [A scholar, fund holder, insurance and tax authority]. PMID- 9340296 TI - Issue dedicated to Sir Ronald Ross. PMID- 9340297 TI - Fish Vaccinology. Oslo, Norway, June 5-7, 1996. Symposium proceedings and abstracts. PMID- 9340298 TI - Image of the month. Appendiceal and sigmoid diverticulosis. PMID- 9340299 TI - A = B < C. PMID- 9340300 TI - [Vascular pedicled transposition of the pisiform bone for treatment of lunate malacia]. AB - In a prospective study we investigated the results of 18 patients with Kienbock's disease stage II as defined by Decoulx, treated with transposition of the pedicled pisiform. In eight cases of minus variance of the ulna, a radius shortening osteotomy was performed. There was an average follow-up of 30 months, X-ray investigations were done every six months after operation. 17 patients had less pain, 14 patients showed an improved range of motion of 30 degrees. Magnetic resonance imaging proved vitalizing of the pisiforme in 16 cases. PMID- 9340301 TI - [Vascular quality of various microsurgical transplants. A histological study]. PMID- 9340302 TI - [Traumatic thrombosis of the distal ulnar artery (hypothenar hammer syndrome) in a golf player with an accessory muscle loop around Guyon's canal. Case report]. AB - Arteriography of a 34-year-old golf player with M. Raynaud symptoms of his left hand revealed filling defects in the digital arteries II to IV fingers associated with corkscrew-like configuration of the ulnar artery in Guyon's space. The preoperative workup including 50 clinical laboratory tests searching for connective tissue diseases, vasculitis, and haematological disorders was without pathological findings; more central embolic sources were excluded angio- and cardiologically. MRI-scan demonstrated an anomalous muscle at the level of the hamate hook located underneath the M. palmaris brevis forming a sling around the ulnar artery. Surgery showed the ulnar artery distal to the anomalous muscle dilated with fibrotic thickening and intraluminal thrombosis. The involved A. ulnaris segment was resected and an interpositional vein graft performed. Histopathologic sections showed fibrosis of the arterial wall with intraluminal thrombosis and elastica fragmentation indicating traumatic genesis (hypothenar hammer syndrome). We suspect intensive golf playing with the grip style and repetitive movements leading to pressure injury of the hypothenar area and the underlying ulnar artery. Contraction of the anomalous muscle belly may have additionally compressed the artery slowing down the arterial flow and accelerating the thrombosis. PMID- 9340303 TI - [A small distal osseous avulsion of the fibrocartilago palmaris of the middle finger interphalangeal joints. Results of conservative therapy]. AB - Chip avulsions of the palmar plate are divided into four degrees (Hintringer). Conservative treatment is preferred for degree I and II and in some cases in degree III. Only a short immobilization for six or seven days is necessary, then the patient will begin with active movement of the fingers. It is important to avoid hyperextension of the joint. Analysis of results in 69 cases with only conservative treatment showed very good results in 72%, good results in 20.4%, and satisfactory results in 4.5%. The results in one case (Hintringer III) was poor. PMID- 9340304 TI - [Possibilities and limits of intramedullary Kirschner wire osteosynthesis in treatment of metacarpal fractures]. AB - An alternative method for treating metacarpal fractures using intramedullary Kirschner-wire pinning is presented. This procedure does not immobilize the metacarpophalangeal joint, thus allowing early motion exercises of the affected hand which is of particular advantage in fractures of the metacarpal neck. Since May 1993, we have treated 33 patients with 37 fractures; the fifth metacarpal was involved in each case. An awl is used to prepare an opening in the cortex for insertion of two or three pre-bent K-wires which are then advanced distally from the base of the metacarpal bone. The hand is immobilized on a plaster splint for one week. Work load is increased after three weeks. With the exception of three cases, our patients achieved free movement of the fingers with anatomical alignment of the fracture site at the time of wire removal. Three cases were re operated upon due to K-wire migration or fracture displacement. Ideal indications for this procedure are distal transverse and short oblique fractures. PMID- 9340305 TI - [Intramedullary osteosynthesis of distal metacarpal fractures with curved wires]. AB - When intramedullary pinning is used to treat metacarpal fractures, as recently described by Forstner (1994) and Foucher (1995), the closed reduction technique developed by Jahss (1938) is applied in the same way as for conservative fracture treatment. It is not always possible to achieve complete anatomical reduction using this closed technique. The intramedullary pinning technique, that we have applied since 1989, involves a Kirschner wire which is bent at one end. Apart from reducing the fracture, the pre-set Kirschner wire serves as a butressing internal fixator. The elastic clamping of the wire acts as an internal wire spring splint, permitting early mobilisation. We have operated on 62 metacarpal fractures using the above-mentioned technique over a period of 6 years until 1995. Anatomic reduction was realized in 50 of 62 fractures. In the follow-up of 32 fractures, we noticed four complications: one infection, two paraesthesias, and one non-union. PMID- 9340306 TI - [Causes for reoperations after osteosyntheses of finger and mid-hand fractures]. AB - 31 patients reoperated after osteosynthesis of a metacarpal or a phalangeal fracture were reviewed over a period of three years. The cause of injury was in 18 cases a crush, in six cases a fall, in five cases a saw injury, and in two cases axial trauma. A postoperative plaster splint immobilisation over a period of at least three weeks was carried out in 29 cases. Reoperation was necessary in most of the cases because of a loss of range of motion and a nonunion. The complication rate was independent of the method of fixation, but did depend on the type of injury. Fractures associated with soft tissue injury were more likely to develop complications. PMID- 9340307 TI - [Implantation of oxidized, regenerated cellulose for prevention of recurrence in surgical therapy of carpal tunnel syndrome]. AB - Recurrences and persistent symptoms of carpal tunnel syndrome after its surgical treatment is most commonly due to inadequacies of the first procedure, recurrent tenosynovialitis and in particular adhesion formation between the median nerve and its surrounding tissue and scars. Because of the encouraging experiences with oxidized regenerated cellulose (INTERCEED) as an absorbable adhesion-barrier in abdominal surgery, we began using INTERCEED in carpal tunnel surgery. In nine patients who underwent division of the flexor retinaculum combined with epineurotomy or synovialectomy, we covered the median nerve with a monolayer of INTERCEED. According to our good experiences concerning handling, compatibility and recovery, the application of INTERCEED might potentially reduce the recurrence rate in carpal tunnel surgery. PMID- 9340308 TI - [100 years tendovaginitis stenosans de Quervain--review of the literature and personal results]. AB - One hundred years ago, Fritz de Quervain first described the surgical treatment of tendovaginitis of the first dorsal compartment. Since then, various ways of treatment have been pointed out. In a series of 72 patients treated surgically, 82% recovered completely. Postoperative complaints included irritations of the superficial branch of the radial nerve. PMID- 9340309 TI - [Rehabilitation of hand injured co-workers from the occupational medicine viewpoint (exemplified by a chassis and montage automobile industry]. AB - The majority (about 25%) of work-related accidents involves injuries to the hands and fingers. Depending on type and severity of the injury, it can have profound consequences to an individual's professional future. Due to this basic insight, hand rehabilitation must include vocational rehabilitation. PMID- 9340310 TI - Removal of PCBs by various white rot fungi in liquid cultures. AB - The ability of Phanerochaete chrysosporium, Trametes versicolor, Coriolopsis polyzona, and Pleurotus ostreatus growing in a mitogen-limited mineral medium (NMM) to degrade PCBs in a commercial, Delor 106 mixture at a concentration of 0.9 ppm was compared. The respective amount of PCBs removed from the fungal cultures within 3 weeks were 25, 50, 41 and 0%. The capacities of the individual fungal species to remove PCBs correlated to some extent with their capabilities of decolorization of NMM agar containing both Poly R-478 or Remazol Brilliant Blue R dyes. Enzyme estimations indicated that both high and relatively stable activities of Mn-dependent peroxidase, Mn-independent peroxidase, lignin peroxidase, and laccase characterized efficient PCB degraders. PMID- 9340312 TI - [Intercorrelation of psychopathological, morphologic, neurophysiologic an psychometric parameters in schizophrenic patients]. AB - The present study was designed to determine the intercorrelation between schizophrenic symptoms, brain morphology, electrophysiological and neuropsychological variables. 44 patients, who met ICD-10 criteria for schizophrenic disorder, were included. At baseline, after 3 and 6 weeks BPRS, CGI, psychometric measurements and QEEG/ERP were performed. A CT scan was performed only at the beginning of the study. Data were evaluated by a multivariate test for data with an inherent structure. One of the most interesting findings is a correlation between BPRS total score and theta EEG power at baseline as well as under treatment. In conclusion, the study suggests the usefulness of multimethodological approaches in order to optimise diagnostic procedures in schizophrenia. PMID- 9340311 TI - [The incidence of neurological disorders in tropical South America. Experience in the Bolivian lowlands]. AB - BACKGROUND: The Chiquitano tribe lives in the southern Amazonas region in Bolivia, remote from larger towns. Data on the epidemiology of neurological disorders are completely lacking. METHODS: A combined prospective-retrospective study was designed to determine the prevalence and annual incidence of major neurological diseases. In an one-year prospective study 1514 individuals (total population 5652) who consulted the general practitioner were interviewed and examined for neurological disturbances. These histories were analysed retrospectively by a neurologist and classified according the diagnosis key of the DGN. RESULTS: During a one-year period (April 1995-March 1996), 139 patients suffering from neurological diseases were seen (one-year incidence and prevalence 2.45%). Cervical and lumbosacral pain syndromes were the most common neurological problems; these were caused by sleeping in hammocks, and by hard agricultural labour. Tropical myositis (12.9%) was very frequent and the most frequent muscle disease. Epilepsy was found in 11 patients and extrapyramidal syndromes in 2 patients. Regarding epilepsy, a high dark rate is assumed because of social and cultural traditions. Strokes are rare, since many risk factors are not present. All cases of meningitis were lethal and clearly demonstrated infrastructural problems. Patients with social diseases (AIDS, drug- and alcohol addiction, injuries caused by violence) were rarely seen. CONCLUSION: In a shrinking world, and with the development of "Tropical Neurology" as a specialised discipline neuroepidemiological data are increasingly important for two reasons. First of all, they sensitise neurologists to this topic, and secondly, they can be used to estimate the need for neurologists serving the Third World's minority populations. PMID- 9340313 TI - [The importance of hypnosis in psychiatry]. AB - This article deals with the significance of hypnotherapy which was almost forgotten in recent years but is currently regaining its position as a special treatment in psychiatry. The historical overview is followed by a discussion of the nature, present forms, indication and risks of hypnotherapy. The most recent evaluation studies are also considered. The results show that the individual forms of hypnotherapy represent an effective and at the same time low-risk form of treatment when the indication is clear and the therapy is administered professionally. It should therefore be used within the framework of both in patient and out-patient psychiatric treatment. PMID- 9340315 TI - [Disturbances of body experience in schizophrenic patients]. AB - Disturbances of body experience in schizophrenia patients occur frequently. They vary phenomenologically and lack exact and distinct definitions. Their theoretical and clinical relevance remains widely unclear. This review summarises the literature on clinically relevant symptoms such as coenaesthesis and body hallucinations, disturbances of pain perception, out-of-body-experiences, dysmorphophobia and self-injuries or self-mutilation. Empirical studies on the concepts of body schema, body concept and body cathexis are reported. Many of these studies have serious methodological shortcomings. The correlation of disturbances of body experience with other psychopathology is considered. Standardised methods for assessing these disturbances are listed. Effects of body oriented psychotherapy have been suggested, but not empirically tested. Finally, the possible relevance of further research in this field is discussed. PMID- 9340314 TI - [The clientel of a psychotherapeutic polyclinic in the light of a documentation base]. AB - A documentation system for use during the diagnostical process with psychotherapy outpatients is outlined, and sociodemographic, medical and psychosocial data of newly admitted patients of a university psychotherapy outpatient department are presented. The population structure is described on the basis of data collected during the first 18 months of the implementation of the assessment system. Data are compared with those of other psychotherapeutical institutions that differ with regard to setting (outpatient vs. inpatient), health care sector (acute care vs rehabilitation) and geographic region. Sociodemographic and medical data are given. Patients' mean age is 33.5 years. Two-thirds of the patients are female. The majority is single and has higher formal education. Slightly more than half of the patients are working. Modes of admittance, somatic and psychological complaints and aspects of illness behaviour are described. Pertaining to diagnostical classification, affective disorders (34%), somatoform disorders (15%) and anxiety disorders (11%) are most frequent. Mean duration of symptoms is 6 years. However, only few patients had prior psychological treatment. Treatments most frequently selected at the time of assessment were psychodynamic psychotherapy (41%), behavioural therapy (24%), and group therapy (10%). When comparing the study population with those of other institutions, differences showed up especially in contrast to rehabilitation clinics, where older, male, and working persons as well as those with somatoform disorders and longer durations of illness are more frequent. PMID- 9340316 TI - [Health government and clinical practice: forgotten institutions?]. PMID- 9340317 TI - [Incidence of pulmonary tuberculosis: application of the capture-recapture method]. AB - OBJECTIVE: To determine as accurately as possible the incidence of pulmonary tuberculosis within Health Area 15 of the Community of Valencia during the period 1990-1993, using the capture-recapture method. METHOD: Descriptive study on the population of Health Area 15 (population: -139.903) divided into 4 large groups according to age (0-14, 15-34, 34-54 and 55+). Data was obtained from the statutory notification system of infectious disease (SNSID) and from the registry of the said area's Hospital Microbiology SERVICE: The main variable under study was the number of cases of pulmonary tuberculosis, both as notified under the SNSID and in terms of cases in the microbiology register in which M. tuberculosis was isolated. The incidence-rates were calculated by age and year of study for both registries employing the capture-recapture method. RESULTS: The mean annual incidence obtained for the SNSID register during the study period was 15.85 cases x 10(-5), and for the microbiology registry it was 23.29 x 10(-5). When the capture-recapture method was employed the mean annual incidence for the study period was 34.81 x 10(-5) (CI 95%: 31.82-39.92). In each of the years studied the number of cases identified was greater for the microbiology register than for the SNSID. Around half the cases of tuberculosis are below 34 years of age, with the larger section, and that having the highest incidence of tuberculosis being the 15 to 34 years age-group. There is no apparent upward trend in incidence rates calculated for this period. CONCLUSIONS: The data from the SNSID system on incidence of pulmonary tuberculosis within Health Area 15 of the Community of Valencia tends to underestimate the true incidence rate. The factual resources of the Hospital Microbiology Service are underutilized, considering the quantity and quality of information it can provide. The capture-recapture method is a good choice of method for measuring tuberculosis incidence. This method merits greater use within the field of epidemiology as much in order to assess the representativeness and thoroughness of surveillance systems as to identify inadequacies in their reporting and localisation of disease outbreaks. PMID- 9340318 TI - [Construction and validation of a severity of illness index of patients hospitalized in clinical areas]. AB - Severity of illness indexes for hospitalized patients are frequently used for the assessment of hospital performance. The purpose here was to develop and validate a quantitative index for clinical areas which could measure severity of illness during hospitalization period and would be easy to obtain from the information contained in a regular clinical chart. The construction included item selection and search for items weights. Experts and literature were consulted and 74 clinical records provided empirical information. The result was the proposal of an index with two alternatives: one quantitative and the other ordinal with four levels of severity. Validation included four aspects of validity, general reliability, interrater agreement and internal consistency. Sixteen specialized physicians assessed content and face validity. One hundred clinical records of discharged patients were used to assess criterion, construct validity and reliability. Results show satisfactory validity in almost all aspects. The reliability coefficient was 0.95, Kappa coefficient was 0.4 for the ordinal index and correlation coefficients over 0.93 for the quantitative one. The index is ready for current applications in this context although some suggestions for future improvements were also included. PMID- 9340319 TI - [Distribution of habits related to health in a female Canarian population]. AB - BACKGROUND: To compare several anthropometric variables, obesity and some life style (tobacco, coffee and alcohol consumption and physical activity during leisure time) in women 45 years old and older. METHODS: From the 1991 electoral roll, we obtained a population of 1221 women aged more than 45 years living in the island of Gran Canaria. A questionnaire and a physical examination including weight and height with light clothes was performed in every woman. Obesity was defined according to the Quetelet index, and weight (in kg) divided by height (in m) at square. RESULTS: Rural women aged more than 45 years old are heavier, have a larger corporal surface, have a higher Quetelet index, smoke less and do greater activity during leisure time than urban women the same age. We found no differences either in alcohol or coffee consumption. CONCLUSIONS: There are statistically significant differences between women that live in rural and urban habitats. Anthropometric variables (weight, corporal surface, Quetelet index) show higher values in rural women than in urban ones. On the other hand, tobacco consumption and sadentarism have a higher prevalence in the urban women compared to rural ones. PMID- 9340320 TI - [Comparison of 2 indexes of prenatal care and risk of preterm delivery]. AB - OBJECTIVES: A new index the adequacy of prenatal care utilization (APNCU) index, has been proposed to provide a more accurate and comprehensive measure of antenatal care use than the widely used Kessner index. To better understand the value of the two above mentioned indexes as predictors of preterm delivery, we examined their ability to predict women who will or will not deliver before 37 weeks of gestation. METHODS: A case-control study was performed, including 207 cases and 381 controls. Prenatal care was assessed on the basis of the two above mentioned indexes, both taking into account the number of prenatal care visits, the date of the first visit and gestational age. Multiple-factor adjusted odds ratios and their 95% confidence intervals were estimated using logistic regression methods. RESULTS: The Kessner index showed a lineal trend with both crude and adjusted for (the APNCU index) estimates of preterm delivery risk. The APNCU index did not show any linear trend with adjusted for (the Kessner index) estimates of preterm delivery risk. To assess whether the Kessner index added explanatory information to the APNCU index (or vice versa), the APNCU index was regressed on the Kessner index (and viceversa), and a set of residuals was computed for both indexes. In logistic regression analyses, the residuals of Kessner added meaningful information to the APNCU index, whereas the residuals of the APNCU index did not add any relevant information to the Kessner index. There results remained unchanged after controlling for several confounders. CONCLUSIONS: A variation in the definition of adequate prenatal care use changes the association between prenatal care and preterm delivery. The Kessner index showed a better ability for discriminating and predicting the risk of preterm delivery. PMID- 9340321 TI - [Problems of the use of the trend test as a dose-response measurement in epidemiology]. AB - BACKGROUND: The objective of the dose-response analysis is to reveal the presence of a progressively response to an increased dose. One of the usual ways of doing so is the carrying out of the so-called trend test. However, the fact that this test may show statistical significance does not necessarily indicate that there is a uniform response between dose and response. This study aims to examine the various situations in which the use of the trend test is not always appropriate as well as given an overview of the use of potential alternatives. METHODS: Five theoretical dose-response scenarios have been created using for supposed case control studies, in which the relationship was not always linear. The results tables obtained have been analysed, leading to the attainment of the Chi-squared trent test, as well as the calculation of the linear component by means of the logistical regression. Subsequently, other alternative methods of trends analysis, such as categorical analysis, the transformation of the findings within the logistical model and the use of risk incrementals have been employed. RESULTS: The test of trends invariably proved to be significant, despite the fact that in four of the theoretical models there did not exist a clear relationship between dose and response. This significant result could lull its readers into proposing a linear dose-response relationship which, in fact, proves false. The alternative use of incremental risk models, or categorical analysis, permits the identification of situations in which there is no relationship. CONCLUSIONS: The trend test should only be employed in cases where the data demonstrates the existence of monotonous, lineal relationship that can be deduced from the results of categorical analysis, the use of incremental risk or the adjustment of various no linear models, although, in this cases, the significance of the test may bring forward no new information. PMID- 9340322 TI - [Validity of homogeneity and independence assumptions in the application of the capture-recapture method with 2 sources of information]. PMID- 9340323 TI - Licensing midwives. PMID- 9340324 TI - [Are there guidelines for treatment of metastasis in laryngeal carcinoma?]. PMID- 9340325 TI - [Occupational cancer]. PMID- 9340326 TI - [Classification, standards and guidelines in otorhinolaryngology, head and neck surgery. Problems in UICC classification of mouth cavity and pharyngeal carcinomas]. PMID- 9340327 TI - [Ear surgery in inner ear deafness. Electronic auditory implants for inner ear deafness]. PMID- 9340328 TI - [The middle ear]. PMID- 9340329 TI - [Cochlear implants]. PMID- 9340330 TI - [Evaluating otolith function with subjective visual vertical discrimination]. PMID- 9340331 TI - [Acoustically optimized middle ear prostheses. New techniques for future research and development of improved implants]. PMID- 9340333 TI - [Guidelines/algorithms of the German Society of Otorhinolaryngology, Head and Neck surgery]. PMID- 9340332 TI - [Laryngeal cancer treatment concept in metastases]. AB - The prognosis of laryngeal carcinoma worsens dramatically with the discovery of primary or secondary lymph node metastases. Thus consistent prophylaxis and therapy of metastases is indispensable. The standard therapy of manifest cervical lymph node metastases is surgery, if possible using the approach of conservative neck dissection. In cases of a lymph node stage N3, i.e. fixated metastases with a diameter greater than 6 cm, individually adapted therapy in necessary because of the extremely poor prognosis. Elective treatment of the N0 neck is surgical, too; however, the site of the primary tumour, possibly infiltrated compartments and, accordingly, invasion of deep lymphatic vessels have to be considered. Special care is necessary if the so-called primitive glottis is invaded (arytenoid region, aryepiglottidian fold, epiglottis). In the case of distant metastases a temporary remission can be obtained with chemotherapy in most cases, but this chemotherapy should not be forced, because there exists no real chance of cure. The trend is towards less aggressive, outpatient schedules of chemotherapy. PMID- 9340334 TI - [Research in psychosocial aspects of ENT tumor surgery (excluding laryngectomy): 4-part analysis of the literature. 3: Results of research]. AB - The available literature on psychosocial aspects of head and neck cancer surgery from 1950 to 1995 was reviewed in four parts. Part 3 is a detailed description of the findings looked at from four different points of view: 1. Restrictions by illness and treatment (dysfunction, mutilation, emotional and social reactions); 2. Coping; 3. Course of illness and treatment (treatment delay, preoperative phase, hospitalization, early and later rehabilitation, tumor recurrences, terminal disease); 4. Relationships between patients and professionals (doctor patient relationships, countertransference). PMID- 9340335 TI - [Evaluating otolith function with subjective visual vertical discrimination]. AB - This report summarizes our experiences with the subjective visual vertical (SVV) as a clinical neuro-otological tool. In the SVV test, patients have to orient a dim light bar in an otherwise dark surrounding earth-vertical, using a remote control. Normal subjects in an upright position did not deviate more than 2 degrees from true vertical. After vestibular neurectomy, the SVV was consistently tilted by some 12 degrees toward the affected ear. Smaller tilts (approximately 7 degrees) of the SVV occurred in patients with spontaneous peripheral vestibular diseases. This shift in SVV disappeared within weeks to months, similar to the spontaneous nystagmus. After stapes surgery slight deviations of the SVV towards the unoperated ear were seen in about 20% of the patients, indicating a slight irritation of the otolith organs. Assessed in an upright position, the SVV thus may be regarded as reflecting tonic otolithic input differences between the two ears. Asymmetries in the shifts of the SVV induced by roll tilts of the gravito inertial vector by eccentric rotations of the subject have been proposed as a test for otolithic sensitivity. In our studies such asymmetries in the shifts of the SVV could not be induced by 26 degrees or 90 degrees roll tilts of subjects towards the affected or healthy ears. A simple clinical test to reveal unilateral otolithic sensitivity (comparable to an otolithic "caloric test") thus still has to be found. PMID- 9340336 TI - [First comparisons with laser vibrometry measurements and computer simulation of ear ossicle movements]. AB - The dynamic behavior of the ossicular chain is very complex and is frequency dependent. To date, this has still not been fully investigated or understood. There remains a lack of measurement procedures to pick up the motion of the ossicles and ear drum simultaneously with sufficient resolution. The presented paper reports simultaneous measurements with laser-Doppler vibrometry at two points of the ossicular chain of cadaver specimens. Motions not observed were derived using mechanical simulation models on a computer and then evaluating appropriate mathematical equations. Using a sound stimulus, the displacement velocities of the umbo and stapes footplate were measured, and the corresponding transfer functions were derived by Fourier transform. Results were used for verification of the computer models. In the current investigations these models were refined and allow for the detailed investigation of the dynamic behavior of the ossicular chain, facilitating the optimal design of passive and active middle ear implants. PMID- 9340337 TI - [Diagnosis and therapy of lymphoid tumors of the orbits]. AB - For the evaluation of neoplastic orbital lesions an interdisciplinary concept is crucial and includes modern imaging techniques as well as rhinosurgical approaches to the orbit. During a 14-year period 6 primary malignant orbital lymphomas and 9 cases of orbital pseudotumors were managed at our department. In most of the cases an exophthalmus with painful swelling of the eyelids, diplopia and a loss of vision were the clinical symptoms. Imaging using CT or MRI revealed homogeneous tissue formations that could not be clearly demarcated from adjacent orbital structures. By a rhinosurgical approach a biopsy was performed of each lesion without surgical complications. Following diagnosis the localized malignant lymphomas of the orbit were irradiated. In one patient systemic spread of tumor required multi-drug chemotherapy. Two patients died as the result of the sequleae of disseminated plasmocytomas. Patients with orbital pseudotumors were treated with systemic steroids. Two patients were also irradiated. No malignant transformation of any orbital pseudotumor was observed in this series. Our experiences show that careful histological evaluation of lymphoid orbital masses is necessary for correct diagnosis, especially since this has great impact on treatment and prognosis. Depending on the localization of a lesion medial orbitotomy by a rhinosurgical approach has proven to be a safe method for performing a biopsy or extirpating tumor. PMID- 9340338 TI - [Electromyography study and follow-up of bilateral recurrent laryngeal nerve paralysis after thyroid gland operations]. AB - Bilateral recurrent laryngeal nerve (RLN) palsies result in dyspnea and often require further surgery to secure an adequate airway for safe respiration. The present study was devised to determine a predictive value of laryngeal electromyography (LEMG) and if it has therapeutic consequences. In a retrospective study of 63 patients with bilateral recurrent laryngeal nerve palsies after thyroid gland surgery we compared the results of LEMG from the thyroarytenoid muscles with the spontaneous recovery rate of vocal cord mobility. A tracheostomy had to be performed in 18 patients because of unsafe dyspnea, while a unilateral lateral cord fixation had to be carried out in 20 patients. Primary thyroidectomies as treatment for benign goiters showed spontaneous normalization of unilateral or bilateral vocal cord mobility in 75% of cases versus 36% after revision thyroidectomies. Single LEMG correctly predicted prognosis in 78% of patients. In all, LEMG was found to be of value in the assessment of bilateral recurrent laryngeal nerve palsies. Findings also showed that a lateral cord fixation should be planned after a 9-month waiting period. When a patient refuses a tracheostomy lateral cord fixation can be performed at an earlier time, with the use of repeated LEMGs to predict whether or not a corrective surgery. PMID- 9340339 TI - [Sector-related grey scale analysis of video ultrasound recorded tongue movements in swallowing]. AB - Diseases of the oral cavity, floor of the mouth, and nervous system can be accompanied by disturbances in tongue movement during swallowing. These disturbances can be diagnosed by videosonography whereby the examiner has to evaluate extensive video documentation of lingual motion. It was the aim of this study to facilitate this evaluation by the application of a reproducible computer assisted quantitative analysis procedure. Video sequences of 56 healthy adults and 19 patients with dysphagias of different etiologies were analysed. A numerical estimation of swallowing movements was carried out in abstraction from the structures imaged (bolus, air, muscles of the tongue, floor of the mouth, hyoid, etc.). Intensity changes of the pixels within previously defined radial image sectors were quantified in relationship to time and depicted as sector curves. The healthy adults demonstrated a characteristic pattern of two motion maxima that appeared within almost all sector curves. These maxima represented bolus transport movements and the reset movement of the tongue. Patients with diseases of the tongue or neuromuscular changes caused by disturbances of the central nervous system showed pathological deviations on videosonography. These appeared as local or general reductions in movement, slow speed motions, repetitive swallowing or unsorted additional movements of the tongue during swallowing. PMID- 9340340 TI - [Diving ranula. A rare disease picture]. AB - The case of a 24-year-old woman with a plunging ranula is reported. Primarily, the patient noticed extensive swelling of the neck, which had developed 3 months previously. The polycystic, firmly attached tumor and the sublingual and submandibular glands of the right side of the neck were surgically removed. Pathological examination showed a retention cyst by means of a plunging ranula. The etiology, pathogenesis, clinical aspect and diagnostic procedures for the plunging ranula are described. Various therapeutic possibilities are discussed in regard to their value. We prefer surgical treatment with a histological examination to verify the diagnosis. The plunging ranula originates on the sublingual gland; therefore, its simultaneous extirpation is recommended to avoid recurrences. In extensive cases, the additional removal of the submandibular gland can be advised. PMID- 9340341 TI - [First isolation of Bilophila wadsworthia in otitis externa]. AB - Bilophila wadsworthia is an obligately anaerobic, gram-negative rod, which was first described in 1989. The natural habitat of this species seems to be the lower intestinal tract where it was recovered from about 60% of the persons investigated. Bilophila wadsworthia is mainly associated with intra-abdominal infections, but the species was also isolated as part of aerobic-anaerobic mixed infections at various extra-abdominal sites. Otological infections due to Bilophila wadsworthia have as yet not been described. In the present paper, we report a case of a patient with postoperative otitis externa following stapedectomy, probably caused by the anaerobes Bacteroides fragilis and Bilophila wadsworthia. PMID- 9340342 TI - [Meningitis. Meningitis, induced by aggressive labyrinthitis]. PMID- 9340343 TI - [Diagnosis and therapy of occult primary tumor with lymph node metastases in the area of the head and neck]. PMID- 9340344 TI - [A case of IGF-II producing liver hepatocellular carcinoma with hypoglycemia]. PMID- 9340345 TI - [A case of chorea-like movement induced during interferon treatment of chronic hepatitis C]. PMID- 9340346 TI - [A case of fluminant Mycoplasma pneumonia and bronchiolitis managed by artificial respiration]. PMID- 9340347 TI - [A case of rapid progressive glomerulonephritis with pulmonary hemorrhage]. PMID- 9340348 TI - [A case of post-kidney transplantation liver failure with hepatitis B carrier state]. PMID- 9340349 TI - [Stress proteins and autoimmune diseases]. PMID- 9340350 TI - [Partial liver transplantation]. PMID- 9340351 TI - [Thrombosis: progress of diagnosis and treatment]. PMID- 9340352 TI - [Thrombosis: relation to blood coagulation mechanism]. PMID- 9340353 TI - [Thrombosis: relation to fibrinolysis]. PMID- 9340355 TI - [Etiology of thrombosis: relationship to vascular endothelial cells]. PMID- 9340354 TI - [Etiology of thrombosis: relationship to blood platelets]. PMID- 9340357 TI - [Diagnosis of disseminated intravascular coagulation]. PMID- 9340356 TI - [Clinical test for early diagnosis of thrombosis]. PMID- 9340358 TI - [Thrombotic thrombocytopenic purpura]. PMID- 9340359 TI - [Hemolytic uremic syndrome]. PMID- 9340361 TI - [Antiphospholipid antibody syndrome and thrombosis]. PMID- 9340360 TI - [Congenital thrombotic tendency]. PMID- 9340362 TI - [Diabetes mellitus and thrombosis]. PMID- 9340363 TI - [Hyperlipidemia and thrombosis]. PMID- 9340364 TI - [Anticoagulant therapy of thrombosis]. PMID- 9340365 TI - [antiplatelet therapy of thrombosis]. PMID- 9340366 TI - [Thrombolytic therapy of thrombosis]. PMID- 9340367 TI - [Thrombin receptor in thrombosis]. PMID- 9340368 TI - [Hemostasis and adhesive proteins in thrombosis]. PMID- 9340369 TI - [Plasminogen and apolipoproteins A]. PMID- 9340371 TI - Articular fracture reconstruction. PMID- 9340370 TI - [Management of thrombosis. Discussion]. PMID- 9340372 TI - [Morphologic and functional diagnostic imaging--vertebrobasilar insufficiency]. PMID- 9340373 TI - Presidential address: high lonesome. From the Southern Association for Vascular Surgery. PMID- 9340374 TI - Genomic medicine. Internet resources for medical genetics. PMID- 9340375 TI - Genetics. PMID- 9340376 TI - [Gastrointestinal bleeding in acute intoxications with non-cauterizing chemicals]. AB - The results of esophagogastroscopy in patients with gastro-intestinal bleeding caused by acute intoxication with noncauterizing chemicals are available. Gastro intestinal bleeding was detected in 96 of 142 patients. Mallory-Weiss syndrome was detected in 45 patients (46.9%), erosions of mucosa of esophagus and stomach- in 27 (28.1%), traumatic lesions of mucosa--24 (25.0%). The curative endoscopic intervention aimed to stop the bleeding and prevent its recurrence was performed in 31 patients. PMID- 9340377 TI - [Intragastric proteolysis after vagotomy in perforative duodenal ulcer]. AB - The examination of intragastric proteolysis in patients operated on the perforative duodenal ulcer has been performed in 80 patients in the period up to 16 years after the surgery. The selective proximal vagotomy was performed in 65 of the patients, the trunk vagotomy--in 15 of the patients. In 55 patients the vagotomy was combined with ulcerrraphy, in 25 patients--with draining operations. The 30-35% decrease of the intragastric pepsin proteolysis was detected in patients after the vagotomy. There was no any signs of restoration of the suppressed proteolytic activity in the time period up to 16 years after the surgery. The draining operations lead to suppression of pepsin proteolysis in long-term period after the surgery and to its substitution with digestion with the pancreatic proteinases. PMID- 9340378 TI - [Potentialities of ultrasound examination in differential diagnosis of benign nodules and breast cancer]. AB - The results of examination of 410 patients with nodular masses of the breast are analysed. Benign tumors were present in 314 (76.6%) of the patients, cancer of the mammary gland--in 96 (23.4%) of the patients. The tumors were classified into 3 types, depending on the intensity of the ultrasound wave behind the tumor. Based on the clinical and morphological analyses it was demonstrated that the presence of the acoustic wave behind the tumor is typical for the invasive cancer of the duct, scirrous carcinoma and lobular cancer. In medullar and mucosous cancer the intensity of the ultrasound wave does not change. In case of intracystic cancer the symptom of "back magnification" is present. The sensitivity of the ultrasound in detection of mammary gland cancer is 88.5%, specificity--96.5%. The analogous parameters of mammography are: 91.0% and 97.8%. In the use of ultrasound control in fine needle aspiration biopsy (FNAB) the sensitivity and specificity of the method increases from 88.5% to 92.7%. If the diagnosis of mammary gland cancer was established with the use of ultrasound and FNAB the additional use of mammography is not obligatory. PMID- 9340379 TI - [Disputable aspects of surgical treatment of acute disseminated purulent peritonitis]. AB - The comparative clinical analysis of effectiveness of two methods of treatment of acute purulent disseminated peritonitis was performed. In the 1st group (63 patients) the semiopen method was used, in the 2nd group (45 patients)--open method was used. The rate of mortality in the 1st group was 68.3%, in the 2nd group--71.1%. The duration of in-hospital period was 53.4 +/- 3.9 days and 52.1 +/- 4.4 days, respectively. A great number of postoperative complications was noted in both groups. In contrast to other authors, no advantages of the open method over the semiopen method were found. PMID- 9340380 TI - [Diagnostic laparoscopy in abdominal injuries]. AB - The results of the treatment of 316 patients with various injuries of abdomen are summarized. 144 (45.6%) of the patients were operated on vital indications (internal bleeding, peritonitis). Additional instrumental examination was necessary in 172 (54.4%) of the patients. Urgent diagnostic laparoscopy was performed in 118 of them. This procedure is indicated in patients with the absence of the symptoms of "acute abdomen" hospitalised in less than 2 hours from the moment of penetrating injury. It is also indicated in case of a doubtful diagnosis in a blunt injury of abdomen, and in alcoholic intoxication. Three groups of the patients were distinguished: 1) patients with indications for urgent surgery (38.1%); 2) patients with indications for conservative treatment (17.8%); 3) patients with indications for out-patient follow-up (44.1%). This policy made it possible to decrease the number of diagnostic laparotomies to 11%, the period of limited disability by 5.8 + 0.3 days, decrease lethality to 6%. PMID- 9340381 TI - [Combination of appendicitis with gynecologic diseases]. AB - 782 patients with combined diseases of abdominal and retroperitoneal organs were operated on from 1970 to 1996. In 321 patients (41.0%) at the age from 15 to 80 years appendectomy in combination with gynaecologic operations were performed. The catarral appendicitis was detected in 202 patients, phlegmonous--in 113, gangrenous--in 6 patients. The rate of inflammatory genital disorders was variable: ovarian cyst rupture (183 patients), suppuration (7 patients), over winding (15 patients), purulent salpingitis (76 patients), ulchanges cyst (6 patients). The extirpation and supravaginal amputation of the uterus were performed in 34 patients. The abdominal exudate was detected in 284 patients. The oblique incision in the ilias region was used in 177 patients. In was extended in 80 patients. The median laparotomy was performed in 64 patients. The combination of appendicitis and gynaecologic diseases are commonly seen in clinical practice. PMID- 9340382 TI - [Intraoperative cholangioscopy]. AB - Intraoperative fibrocholangioscopy (FCS) was used in 446 patients with cholelithic disease and acute cholecystitis. 399 (89.5%) of the patients were hospitalized urgently. The use of FCS in patients with choledocholithiasis, obturative jaundice, cholangitis and pancreatitis is analysed. The safety and diagnostic accuracy of the method are proved. The use of FCS has provided significant improvement of the immediate and long-term results of the biliary tract surgery. PMID- 9340383 TI - [Influence of regional lympho-immunostimulation and infra-red magnetolaser therapy on structure of lymph nodes of lesser omentum after gastric surgery]. AB - The influence of regional lymphoimmunostimulation and low-intensive laser irradiation on the structure of lesser omentum lymphatic nodes short time after vagotomy and gastrotomy were studied. The lymphostimulation with T-activin decreases signs of congestion in lesser omentum lymphatic nodes and promotes the process of immunocompetent cells' migration, increases the resorptive capacity of the lymphatic system. Laser irradiation, combined with lymphoimmunostimulation, provides the protective action on the structure of lesser omentum lymphatic nodes, and stimulates differentiation of immunocompetent cells. The action of lymphoimmunostimulation is potentiated by infra-red laser irradiation in their combined use. PMID- 9340384 TI - [Combined treatment of stomach neoplasms]. AB - The results of treatment of 394 patients with gastric cancer were analysed to compare the effectiveness of surgical and two variants of combined treatment (preoperative irradiation with intensive-concentrated IKK method and dynamic DFD dose-fractioning). The advantages of combined treatment over surgical treatment are demonstrated. The 3-year survival in the combined treatment is 70.2%, in surgical treatment 34.5 +/- 6.2%. The advantages of SDF preoperative irradiation over IKK irradiation are revealed, that is proved by the increase of the 3-year survival rate (76 vs. 56.7%), and decrease in the rate of recurrence from 50 to 27.3%. The addition of metronidazol leads to increase of anticancer effectiveness, that is proved with the examination of tumor pathomorphosis and the rates of survival. PMID- 9340385 TI - [Significance of greater omentum anatomic structure]. AB - The examination of the greater omentum was performed in 102 cadavers. Most frequently the quadrangular shape of omentum was found, less frequently- triangular and appendicular shape. The anterior duplicature of the omentum receives blood supply from the right gastro-omental artery, that often has 5 omental branches. The posterior duplicature is blood supplied by spleno-omental artery, that creates the inferior arterial arch. The methods of lengthening of omentum are worked out. Satisfactory filling of the created omental flaps was proved by arteriography. PMID- 9340386 TI - [Treatment of purulent postoperative wounds with collagen and immobilised gentamycin]. AB - The results of treatment of purulent postoperative wounds in 184 patients are discussed. All the patients were operated on urgently. Various destructive diseases of the abdominal cavity organs, complicated with peritonitis, were the indication for the surgery. The abdominal cavity was drained through laparatomic wound. The wounds became suppurative with subsequent diastasis of the wound edges. Microbiologic, cytologic and clinical tests were used to estimate effectiveness of the treatment with the use of collagen sponge with immobilised gentamycin. The method has provided fastening of the healing process and shortening of in-hospital stay. PMID- 9340388 TI - [Choice of optimal anesthesia in outpatient treatment of varicose veins]. PMID- 9340387 TI - [Functional outcomes of legs obliterative endarteritis treatment by Ilizarov's method]. AB - The method of Ilizarov was used for the treatment of 94 patients with obliterative endarteritis (stage II-IV). The significant improvement of functional status of the patients has been achieved as well as the improvement of peripheral blood circulation. The rate of positive results in the long-term follow up period was 57-70%. The rate of improvement depended on the functional status of vascular system. The method of Ilizarov is indicated in patients without occlusion of iliac and femoral arteries, and the pressure index more than 0.6; the peak blood flow index more than 3.0. PMID- 9340389 TI - [Antibiotic prophylaxis of postoperative infective complications in patients with rectal and colonic cancers]. AB - The results of operative use of clindamycin and netilmycin (1st group), piperacillin (2d group), in comparison to the standard protocol (carbenicillin and its combination with gentamicin) in 103 patients operated on rectal and colonic cancer are discussed. The rate of postoperative complications was 2.3% (1st groups), 5.3% (2d groups) vs. 37.1% (control group). The authors advocate the proposed protocols of antibacterial prophylaxis of rectal and colonic cancer. PMID- 9340390 TI - [Long-term results of giant hernias treatment after obstetric and gynecologic surgeries]. AB - The author studied in 311 women long-term results of treatment of giant hernias, that were caused by urgent (52.4%) and planned (47.6%) obstetric and gynecologic operations. The original lavsan prostheses were used to cover the suprapubic abdominal wall defects that provided good long-term results with no cases of relapses, good functional and cosmetic effects, and complete restoration of capacity for work. PMID- 9340391 TI - [Non-reflux esophago-gastric anastomosis]. AB - A new method of performing esophago-gastric anastomosis in proximal gastric resection is proposed. It is based on a circular enveloping of the anastomotic zone with the "spurs" of the greater curvature of the stomach. The method was designed and experimentally tested on nonfixed cadavers, as well as on the 60 dogs. Esophago-gastroscopy, histological examination of the anastomotic zone and mucosa of esophagus were used to evaluate long-term results. The method was used in 8 patients with proximal gastric resections. Good permeability of the esophagus and lack of signs of anastomotic insufficiency were found. The method is advocated for the use in clinical practice to provide safe peritonization of anastomotic zone and prevent gastroesophageal reflux. PMID- 9340392 TI - [Method of surgical treatment of hiatal hernia]. AB - A method of hiatal hernia surgery providing an opportunity to decrease postoperative complications has been proposed. The method eliminates the development of a pathologic hiatal narrowing during diaphragmocrurorraphy. The method has been used in 65 patients. The advantages of the method are demonstrated. PMID- 9340393 TI - [Expended radical surgery of sigmoid cancer with spreading to the bladder]. PMID- 9340394 TI - [Fluorescent endoscopy in diagnosis of stomach neoplasm]. PMID- 9340395 TI - [Zollinger-Ellison syndrome]. PMID- 9340396 TI - [Contact lenses. 1: Contact lens associated complications]. AB - The contactology as frontier between ophthalmology and eye optics requires special attention by part of ophthalmologists. Contact lenses influence the anatomy, the physiology and the immunological responses of the external eye. The corneal metabolic disorders of contact lens wearers are among others, the result of a diminished oxygen supply to the cornea. These disorders are influenced by contact lens deposits, tear film dysfunction and intrinsical characteristics of contact lenses. Sensibilisation and allergic reactions are result of the wearing of contact lenses. Even toxic reactions, as a result of the disinfection systems used, may be observed Ophthalmologists must be able to recognize the contact lens induced diseases and to advice their patients properly. Therefore, regular ophthalmological control examinations are indispensable. PMID- 9340397 TI - [Eye damage caused by ultraviolet sunlight. The sun--blessing or curse?]. PMID- 9340398 TI - [Physiological protective mechanisms of the eye]. AB - The physiological mechanisms for the protection of the eye are only partly known. It becomes increasingly evident that the eye is supervised by and included in the immune system of the body. The physiological protection system of the eye consists of a mechanical and an immunological component. The lacrimal gland plays a key role for the immune system of the anterior segment. It is part of the MALT system, and the characterisation of the special ocular components of this system including possible homing receptors-will give important informations. Research of the ocular immune system has disclosed various special features, like the anterior chamber associated immune deviation (ACAID). The distribution of antigen presenting cells and its localization, but also the distribution of T- and B lymphocytes is an important factor for limiting ocular inflammation. The phenomenon of 'apoptosis' seems to have an important role in maintenance of the ACAID. The cornea is nearly free of cells, and infiltrating antigen presenting cells may prevent ACAID. Research regarding mechanisms which are used to tuned these various cells will give answers to the questions how the cornea contains its optical transparency, how corneal transplant rejection works, how the eye participates in systemic disorders and may also define the role of a possible dysfunction of antigen presenting cells of the retinal pigment epithelium in senile maculopathy. PMID- 9340399 TI - [Current status of studies on development of myopia]. PMID- 9340400 TI - [Pseudoexfoliation syndrome in patients with retinal vein branch and central vein thrombosis]. AB - BACKGROUND: Pseudoexfoliation syndrome is a frequent cause of secondary open angle glaucoma due to deposition and in situ-production of abnormal extracellular matrix proteins in the trabecular meshwork. Glaucomas itself are a known risk factor for retinal vein thrombosis. In this retrospective study the prevalence of pseudoexfoliation syndrome in patients with branch and central retinal vein thrombosis was investigated. PATIENTS AND METHODS: The records of 332 unselected patients with branch retinal vein thrombosis and 159 with central retinal vein thrombosis, which were seen in our laser out-patient clinic between January 1990 and July 1996, were analysed retrospectively. RESULTS: 6.0% of patients with branch and 6.9% of patients with central retinal vein thrombosis revealed pseudoexfoliation syndrome. The full picture of pseudoexfoliation syndrome was seen in 4.2% of patients with branch and in 4.4% of patients with central retinal vein thrombosis. The prevalence of pseudoexfoliation syndrome increased with age. Twenty percent of patients with branch and 27% of patients with pseudoexfoliation syndrome and branch or central retinal vein thrombosis. The branch retinal vein thrombosis was significantly more often localised at the disk in patients with glaucoma and in patients with pseudoexfoliation syndrome. CONCLUSIONS: Pseudoexfoliation syndrome is a risk factor for retinal vein thrombosis. Pathogenetically the secondary open-angle glaucoma caused by pseudoexfoliation syndrome could be causative for the venous thrombosis. PMID- 9340402 TI - [Optic nerve neuritis after 50 years of age: symptom of a previously undiagnosed multiple sclerosis]. AB - BACKGROUND: The onset of multiple sclerosis after the age of 50 years is rare. We report four patients with optic neuritis and oligoclonal bands or increased IgG production in the cerebrospinal fluid. Three of them subsequently developed clinically definite multiple sclerosis. PATIENTS: Four apparently healthy women, 62, 56, 50 and 50 years of age, presented with progressive visual loss, three patients in one eye and one patient in both eyes. The diagnostic work-up revealed no findings indicating an ischemic process, an infectious, systemic inflammatory, or neoplastic disease. Examination of the cerebrospinal fluid revealed oligoclonal bands and/or increased production of IgG. Visual acuity recovered during the following weeks to 8 months. Three of the 4 patients subsequently developed additional neurologic signs compatible with multiple sclerosis. CONCLUSION: Optic neuritis can be the presenting sign of multiple sclerosis even after the age of 50. PMID- 9340401 TI - [Rapid diagnosis of infectious endophthalmitis using polymerase chain reaction (PCR): a supplement to conventional microbiological diagnostic methods]. AB - BACKGROUND: Endophthalmitis is, although relatively rare, a serious intraocular infection, which could result in a loss of visual function. Therefore, the rapid diagnosis and initiation of the appropriate treatment is of critical importance. To date, approximately 60 percent of eyes with a clinical diagnosis of infectious endophthalmitis show a positive microscopic or culture result. By using the very sensitive polymerase chain reaction this number might increase. PATIENTS AND METHODS: In a series of 12 eyes with infectious endophthalmitis we have performed microscopic investigations, diagnostic culture and polymerase chain reaction in aqueous humor and vitreous in order to detect the infectious agent. RESULT: Microscopic investigations showed a positive result in the vitreous of 3 eyes. This number improved to 6 eyes using culture media. Significant less positive results were obtained in the aqueous humor. The infectious agent could be detected in the aqueous humor in all 12 eyes and in the vitreous in 9 eyes by PCR. Only in 2 eyes with a delayed endophthalmitis the vitreous was negative. CONCLUSIONS: The detection of the infectious agents was more successful using PCR compared to conventional microbiological tests. In particular, for the diagnosis of delayed endophthalmitis PCR proves to be very superior. In all cases of delayed endophthalmitis the pathogen could be detected in the aqueous humor. PMID- 9340404 TI - [Frontalis suspension with "expanded polytetrafluoroethylene (ePTFE) strips" in congenital ptosis]. AB - BACKGROUND: The Frontalis Suspension Technique is indicated in cases with minimal or no levator function. At the beginning sutures were used as sling material and after further modifications suture material was replaced by autologous or homologous fascia lata. In the last years ePTFE has proved to be a very suitable sling material. PATIENTS AND METHODS: Since ePTFE is very biocompatible, it was used for this modified frontalis suspension technique. The anterior tarsal surface is exposed and a small tunnel is created between the skin incision and the second incision superior to the brow. An 0.3 mm thin ePTFE strip is induced into the tunnel and connects the upper lid with the frontalis muscle. The ePTFE Soft Tissue Patch must be exactly adapted to the tarsus and has to be deeply sutured to the frontalis muscle below the brow incision. Since 1994 17 modified frontalis suspension procedures have been performed on 14 patients. RESULTS: The functional and cosmetic result were good in nearly all patients. No implant had to be removed during the follow up period. DISCUSSION: The new technique of frontalis suspension using a ePTFE strip guarantees a regular upper lid lifting by the axial and direct connection of the anterior tarsal surface with the frontalis muscle. PMID- 9340403 TI - [Development of corneal sensitivity after phacoemulsification with scleral tunnel incision]. AB - BACKGROUND: after anterior segment surgery the anatomical structures and in particular corneal metabolism and corneal trophic are reduced. The corneal wound healing might be monitored by measuring corneal sensitivity. MATERIAL AND METHOD: In a prospective study of 1 year corneal sensation of 26 patients, age = 72.49 +/ 11.09 years, after phacoemulsification with scleral tunnel is examined. Corneal sensitivity is measured with the Draeger aesthesiometer in the corneal center, in the 6 o'clock and 12 o'clock position before and 4 weeks, 4 months and 1 year after the operation. RESULTS: The preoperative corneal sensitivity thresholds before the operation are with 11.61 +/- 10.49 x 10(-5) N in the 12 o'clock position, in the center with 2.43 +/- 2.57 x 10(-5) N and in the 6 o'clock position with 11.86 +/0 10.37 x 10(-5) N normal. Even 1 year after surgery corneal sensitivity is markedly reduced with 257.08 +/- 306.71 x 10(-5) N, 107.77 +/- 246.76 x 10(-5) N and 82.81 +/- 140.77 x 10(-5) N in the 6 o'clock position. CONCLUSION: Even 1 year after phacoemulsification corneal sensitivity is definitely hyposensitive in the center and descretely hypotensitive in the periphery at the 6 o'clock. PMID- 9340405 TI - [Quality of vision screening in childhood]. AB - BACKGROUND: In Germany the paediatricians use three different visual acuity tests (Pictures, Rodenstock-Test, H-Test). We investigated the effectiveness of these procedures. SUBJECTS: In this study 303 children were referred by paediatricians to the study group in the Cologne Augenklinik. They were fully examined by an orthoptist and an ophthalmologist (gold standard). RESULTS: From these data the sensitivity and specificity of the tests used by the paediatricians were calculated. Only the H-Test proved to be satisfactory. CONCLUSION: The actual situation of screening for amblyopia by paediatricians in children aged 3 1/2 to 4 is unsatisfactory in Germany. Paediatricians and ophthalmologists should use a similar test for visual acuity. Continuing education of medical personnel is desirable to ensure an adequate quality of screening. PMID- 9340406 TI - [Modified radiation administration in Er:YAG laser ab externo sclerostomy]. AB - BACKGROUND: Ab externo sclerostomy enables the restrained aqueous humour drain via a microgram wide corneoscleral canal into the subconjunctival space being an alternative surgical method in glaucoma therapy. The tissue ablative infrared laser radiation (2.94 microgram) was applied until now via a quartz fiber manually pushed forward in permanent tissue contact. Our aim was the development of a new applicator tip without manual feed in order to achieve a reduction in times for construction and maintenance of the former tip as well as in the risk of an infection and mechanical damage of intraocular structures. MATERIAL AND METHOD: In pig eye experiments, the laser parameters were varied in histological and scanning electron microscopic evaluation (pulse energy: 10-60 mJ, repetition rate: 5-17.5 Hz, combinations of constant power: 200 mW). Their influence on the canal shape and the morphology of the internal ostium has been proven in special tests. RESULTS: The new development of the applicator tip (core diameter: 550 microgram) enables a conical profile of the beam (diameter 500-150 microgram) within a distance of 2-3 mm, now without pushing the fiber forward ("fixed fiber"). Smooth canal surfaces, a small homogeneous necrosis zone (25 microgram) and only few tissue ruptures have been seen histologically and on SEM. The correlations of ablation rates/-speed as well as canal characteristics are more significant to energy densities than to repetition rates. First clinical application was technically successful. CONCLUSIONS: The new applicator tip optimizes the method of Er: YAG-laser ab externo sclerostomy from the technical and surgical point of view. PMID- 9340407 TI - [Bilateral candida endophthalmitis in 2 i. v. drug-dependent patients with oral L methadone substitution]. AB - BACKGROUND: Controlled therapy programs for former i.v. heroin abusers supervised by physicians offer levomethadone (L-Polamidon) as an oral substitute for heroin to allow social stabilisation of the addict. As well they reduce the risk of bloodborne infections (e.g. HIV and hepatitis) by giving up "needle sharing". Assuming that i.v. drug abuse is one of the major risk factors for Candida endophthalmitis in young, otherwise healthy patients, in case of strict oral substitution no onset of Candida endophthalmitis should be expected. CASE REPORT: The clinical courses of two HIV-negative and primarily i.v. heroin addicted young male white patients are presented. While participating in an exclusively controlled, oral program of methadone substitution for several months, both patients developed bilateral Candida endophthalmitis. CONCLUSION: A persisting i.v. abuse, either of heroin or of L-methadone, parallel to the supposingly strict orally applied substituting drug L-methadone has definitely to be suspected leading to the Candida endophthalmitis. PMID- 9340408 TI - [Successful irradiation of ciliary processes with electrons in therapy refractory aphakic glaucoma]. AB - BACKGROUND: Successful radiation was already reported by Genee and Weitzel (1968): Electrons from a Betatron were used for irradiation of the ciliary epithelium to lower aqueous humor secretion. To our knowledge, no other paper on this topic has been published so far. PATIENT: A 26-year-old man had history of concussion trauma and cataract surgery in childhood. In adult age, cataract surgery and retinal operations were followed by numerous surgical procedures and maximal medical treatment because of a secondary glaucoma. The patient refused further surgical and medical treatment with the exception of the administered systemic carbo-anhydrase inhibitor. Because of severe side-effects caused by the acetozolamide, irradiation of the ciliary processes with electrons, emitted from a linear accelerator, was performed. The total amount of 12 Gy both on the nasal and on the temporal side of the ciliary body was fractionated in several sessions. RESULTS: During a follow-up of 22 months without medication, the mean value of i.o. pressure was 22.1 mm Hg. The patient is doing well, corrected VA is 0.8. CONCLUSION: Irradiation of the ciliary processes with electrons for lowering aqueous humor secretion was effective in an otherwise non treatable case of glaucoma. PMID- 9340409 TI - [Unilateral trigeminal nerve hypoplasia]. AB - BACKGROUND: Isolated unilateral corneal anaesthesia represents a very rare clinical entity. The underlying cause may be a hypoplasia of the trigeminal nerve. HISTORY AND CLINICAL FINDINGS: A 7 year old otherwise healthy boy presented with mixed conjunctival injection of the left eye, fluorescein-positive punctuate epithelial keratopathy of the cornea and a central corneal ulcer OS. History revealed intermittent, painless redness of the left eye since the age of 4. Trigeminal defects caused by trauma or infection could be ruled out. Tyndall's phenomena was positive. There was no corneal sensitivity on the left side and facial sensitivity was reduced in all branches of the trigeminal nerve. All other ophthalmologic examination results were normal. Magnetic resonance tomography showed a hypoplastic left trigeminal nerve. Mesenchymal syndromes could be ruled out by neuropediatric examination. THERAPY AND CLINICAL COURSE: Treatment with prednisolone and antibiotic ointment and eye patching were performed. The ulcer healed completely and artificial tear substitution was given for prophylaxis. Follow-up examinations after 4 and 6 years showed no signs of inflammation. Biomicroscopy showed only mild fluorescein-positive corneal epitheliopathy. CONCLUSIONS: In cases with painless intermittent keratoconjunctivitis, sometimes associated with corneal ulceration, in early childhood, one should consider acquired or congenital trigeminal anaesthesia. This condition requires life-long corneal ulcer prophylaxis and regular ophthalmologic exams. PMID- 9340410 TI - [Spontaneous hematocornea after keratitis of various etiology]. AB - BACKGROUND: Corneal blood staining may occur as a serious complication of persisting hyphema. In our two cases the corneal blood staining is a result of direct bleeding in the corneal stroma. HISTORY AND SIGNS: A female patient presented with a corneal blood staining through rupture of reopened vessels in interstitial keratitis of congenital syphilis after physical effort with high blood pressure. The second patient presented with a blood staining caused by a vessel errosion after corneal ulceration. THERAPY AND OUTCOME: The clearing of the blood staining is thought to be a result of the phagocytic action of the keratocytes and from a diffusion of hemoglobin into the conjunctival circulation and the anterior chamber (2). The therapeutic efforts are directed toward prevention of corneal blood staining. At the first sign of microscopic blood staining of the cornea a surgical evacuation of hyphema is necessary. In our two cases there wasn't a hyphema, so it was only possible to treated the corneal ulcer, entropion and systemic hypertension. CONCLUSION: The clinician has to wait for a spontaneous clearing, although it may take 2 or 3 years or more. In this case a penetrating keratoplasty is indicated. PMID- 9340411 TI - [Enophthalmos caused by orbital metastasis of breast carcinoma]. AB - BACKGROUND: Orbital metastatic disease usually leads to exophthalmos but rarely to enophthalmos. We report a case of a metastasis causing enophthalmos. PATIENT: A 68-year-old woman had mastectomy for breast cancer six years prior to presentation. She complained of double vision when looking sideways. The right eye showed a motility reduction in all directions and a slight ptosis. She had 4 mm enophthalmos, and the eyelids were sunk into the orbit. There were no signs of optic nerve damage. Magnetic resonance imaging showed a retrobulbar mass surrounding the optic nerve and infiltrating the muscles. The space of the orbital fat was reduced. A biopsy confirmed the diagnosis of metastatic breast carcinoma. Histologically, the connective tissue was infiltrated by lymphocytes, and the nuclei of the tumor cells where aligned in a linear "indian file" pattern. 30% of the tumor cells contained the estrogen-receptor protein, 40% the progesterone-receptor protein. The CA-15/3 and CEA levels were elevated. The patient underwent orbital radiation with 50 Gy. During the following 2 months, the enophthalmos increased to 6 mm. DISCUSSION: We suggest the following hypothesis as the cause of enophthalmos in orbital metastases: The tumor growth goes along with fibrosis. Subsequent shrinkage of the connective tissue pulls the eye back into the orbit. The ensuing elevation of tissue pressure leads to atrophy of the retrobulbar fat. The increase of tumor volume is too slow to compensate for the fat atrophy. Slowly progressive enophthalmos with reduced motility is nearly pathognomonic of metastatic scirrhous breast carcinoma. In rare cases, a diffusely infiltrating carcinoma of the gastrointestinal tract may cause a similar picture. PMID- 9340412 TI - [Intrapalpebral migration of a form stable contact lens: a rare complication in contact lens practice]. AB - PURPOSE: To demonstrate a rare complication in contactology. CASE REPORT: A 26 year-old female had "lost" her hard contact lens in the right eye 2 years previously. Afterwards, new contact lenses were not tolerated. The patient had a slight intermittent epiphora but no further complaints. She consulted her ophthalmologist for new spectacles. At presentation, there was a firm tumour without signs of inflammation in the right upper lid area. When the lid was everted a hard contact lens was found within the tarsal plate which could be easily removed in the operating room. Microbiologic investigation disclosed no bacteria. Histology showed a circumscribed papillary reaction and a chronic non specific inflammation with few eosinophils and no giant cells. Two weeks later the lens-related cavity was only slightly filled up by granulation tissue. CONCLUSIONS: After "loss" of a contact lens superior dislocation and finally tarsal implantation should be kept in mind. The process of contact lens migration reveals some interesting features: 1. Generally, it causes only minor symptoms though it may last for years. 2. Bacterial contamination rarely occurs. 3. Histologically, the inflammation is often mild or even absent. Eosinophils and giant cells are of minor or no importance indicating that allergy and giant-cell reaction play no significant role. 4. The lens related cavity probably heals slowly, possibly because of a (partial) epithelialization. PMID- 9340413 TI - [The effect of light with various wavelengths and impulse times on nocturnal suppression of N'acetyltransferase activation by serotonin in the pineal gland of the chick]. AB - PURPOSE: Chick pineal gland synthesise melatonin in circadian rhythm, with peak values in the dark phase of an imposed light-dark illumination cycle. Light is the most important environmental factor regulating the melatonin-generating system in this gland. Exposure to light causes a dramatic decline of the night time levels of the melatoninergic parameters. Effect of white and monochromatic lights of various wavelength on the night-time pineal gland serotonin N acetyltransferase (NAT) activity were examined in chicks. MATERIAL AND METHODS: Male chicks (white leghorn: 3-4-week old) were used. They were purchased on the day of hatching. All animals were offered ad libitum access to standard food and water, maintained under an ambient temperature of 27 +/- 2 degrees C (chick), 60 +/- 5% humidity, and exposed to 12 hr light: dark illumination cycle for a minimum of 8-10 days before experiments. The daytime light intensity at the surface of the animals' cages was about 150 luxes. Each experiment was done at least twice. At the beginning of the fourth hour of the dark phase of the light dark illumination cycle groups of chicks (four animals/group) were exposed to either white or monochromatic light for 5, 10, 30 or 60 min, and then killed by decapitation. In another set of experiment birds were illuminated for 5 min, returned to darkness for additional 5, 15, 60 or 120 min, and then decapitated. Decapitation was done quickly under dim red light (2 lux). Pineal glands were isolated and frozen on dry ice. The exposure of an animal to light took place in 25 x 21 cm white plastic chamber. Light produced by 5 W 14 bulbs (Osram) was passed through a cotton filter or filtered with glass, narrow band interference filters, 7 nm half-peak band-width. The spectral wavelength analysis for each interference filter was performed with the aid of Diode-Spectrophotometer, and the irradiance of the light of the three used wavelengths was measured with YSI Radiometer. The estimated peak wavelengths (Imax) of the filters were: 434 nm (blue), 548 nm (green) and 614 nm (red). The NAT activity was determined in supernatants of tissue homogenates with the radioisotopic method of Steinlechner with modifications by Nowak. RESULTS: Significant decrease of the night-time NAT activity of the chick pineal gland was observed after a 5-min pulse of either white or green light. In birds illuminated by blue light a marked reduction (by about 22%) in the pineal NAT activity was found after 10-min exposure, whereas in the group of animals that were exposed to red light a significant decline of the enzyme activity in pineal gland was produced by a 30-min pulse. The enzyme activity started to increase after 15 min in the dark and reached the control values by 1 hr (red light) or 2 hrs (blue and green lights). Two hrs and 5 min after the exposure to white light (5 min light and 120 min dark) pineal NAT activity was still significantly lower (by 16%) than the activity found in the dark control group. CONCLUSION: Suppressive effect of monochromatic (but not white) light on NAT activity was completely reversible in the pineal gland of chick. PMID- 9340414 TI - [Tissue plasminogen activator in treatment of fibrinous membranes after cataract surgery]. AB - The aim of our study is estimation of the efficacy of recombinant tissue plasminogen activator for the treatment of fibrinous membranes following cataract extraction. PATIENTS AND METHODS: The fibrinous membranes were formed, despite conventional antiinflammatory therapy, in 17 eyes after ECCE with IOL and in 3 eyes after ECCE without IOL. We injected the solution of recombinant tissue plasminogen activator-Actilyse (Boehringer Ingelheim) into the anterior chamber of 20 eyes after cataract surgery. Simultaneously, topical corticosteroid and cycloplegic therapy was continued. Routine follow-up included slit lamp examination, ophthalmoscopy and intraocular pressure measurement. RESULTS: Injection of the solution of tissue plasminogen activator into anterior chamber of 20 eyes resulted in complete dissolution of the fibrinous membranes in 18 eyes within 1 day. In 7 eyes, within a few days after injection, fibrinous membrane recurred. In four of this cases durable effect of fibrin dissolution was obtained after multiple tPA injections. Finally, expectable stable result was obtained in 15 of 20 eyes. CONCLUSIONS: Results of our study confirm high efficacy of recombinant tissue plasminogen activator in the treatment of fibrinous membranes after cataract surgery. Short term follow-up and small number of cases motivates further continuation of observation. PMID- 9340415 TI - [Use of 5-fluorouracil shortly after trabeculectomy]. AB - PURPOSE: To present own results of treatment wit 5-FU in the cases with high intraocular pressure and increased cicatrization shortly after trabeculectomy. MATERIAL AND METHODS: Subconjunctival injections of 5-FU were applied in 12 patients in whom, immediately after trabeculectomy, intraocular pressure was above 22 mmHg and conjunctiva on the filtering bleb was thickened. RESULTS: Normalization of the intraocular pressure and good function of the filtering bleb was achieved in 83.3% of patients. PMID- 9340416 TI - [Corneal astigmatism after tunnel incision for cataract extraction]. AB - PURPOSE: To evaluate the development of astigmatism after cataract extraction depending on the kind of incision MATERIAL AND METHODS: Astigmatism was examined in 107 eyes of 92 patients, who underwent cataract extraction, mostly with IOL's implantation. In 52 eyes phacoemulsification with tunnel incision, measuring 5 mm in 27 and 8 mm in 25, was performed. In the control group of 55 eyes incision with scleral flap measured 2/5 of the corneal circumference. In cases with 5 mm incisions no sutures were used, in 8 mm incisions wounds were closed with single sutures and in control group the double continuous sutures were applied. Astigmatism was measured in two days, one week, four weeks and 3 months after surgery. RESULTS: Post-operative astigmatism decreased gradually and after 3 months was average 0.14 D in the eyes without sutures, 0.68 D in those with single ones and 1.78 D in the control group. PMID- 9340417 TI - [Surgical management of intraocular lens dislocation into the vitreous cavity]. AB - PURPOSE: To present our experiences in the management of intraocular lenses (IOLs) dislocated into the vitreous cavity. MATERIAL AND METHODS: The authors report three cases with posteriorly dislocated IOLs in which pars plana vitrectomy techniques and limbal incision were used for IOLs removal. In one of these eyes perifluorocarbon liquid was injected after vitrectomy to float the IOL off the retina to the retropupillary space and transclerally sutured ciliary sulcus fixation IOL was implanted simultaneously. Anterior chamber IOLs were implanted in the two other cases. RESULTS: Best corrected postoperative visual acuity was between 5/5 and 5/10 in all eyes. No important complications were observed. CONCLUSIONS: Application of vitreoretinal microsurgical techniques allows for the safe removal of dislocated IOLs. Primary or secondary IOL re implantation gives quick visual rehabilitation. PMID- 9340418 TI - [Evaluation of posterior capsule opacification after extracapsular cataract extraction with and without implantation of intraocular lens]. AB - PURPOSE: The aim of the study was evaluation of the rate of posterior capsule opacification after extracapsular cataract extraction with and without implantation of intraocular lens. MATERIAL AND METHOD: The clinical material comprised 77 patients operated on in the years 1990-1994. Only the cases in which visual acuity exceeded 5/10 were included into the study. 107 eyes were examined, among them 75 after implantation of posterior chamber intraocular lens. RESULTS: Opacification of the posterior capsule occurred in 37 eyes, but only in 25 of 75 eyes in the patients with implantation of the artificial lens. We found that following extracapsular cataract extraction without implantation of intraocular lens, opacification of the posterior capsule occurs more frequently, especially during 1-2 years after the surgery. CONCLUSION: Cortical-nuclear cataract and not much advanced age are factors predisposing for greater rate of opacification of the posterior capsule after extracapsular cataract extraction. PMID- 9340419 TI - [Surgery of orbital tumors with the Kronlein-Berke-Reese method in material from the Ophthalmology Clinics in Balystok]. AB - PURPOSE: To analyse the results of surgical treatment of tumors located in the posterior part of the orbit. MATERIAL AND METHODS: 12 patients (5 women and 7 men, aged 10-70 years) with tumors located in the posterior part of the orbit, operated on in the years 1985-1995 in Ophtalmology Clinic in Bialystok with the use of Kronlein-Berke-Reese lateral orbitotomy method. Tissues of removed tumor were evaluated histopathologically. RESULTS: In 7 cases tumor was removed completely, including 3 cases with simultaneous optic nerve removal. In 3 other cases tumor was removed partially, whereas in 2 cases eventration of the orbit was necessary. Persistent postsurgical defects of the eyeball mobility were not found in any of the patients. In 2 patients with partially removed tumor a constant blepharoptosis was observed, which required further operation. Visual acuity in patients with preserved optic nerve remained unchanged in 6 out of 7 cases. In 1 case-completely removed cavernoma-visual acuity decreased from 5/7 to 5/16. CONCLUSIONS: Lateral orbitotomy enables successful removal of tumors located in the posterior part of the orbit. Moreover, this method is relatively safe, especially in comparison with cranial approach to tumors badly located in the orbit. PMID- 9340420 TI - [Long-term observations of ocular toxocariasis in children and youth]. AB - PURPOSE: To evaluate the clinical status and ELISA test changes in a group of children with ocular toxocariasis. METHODS: We enrolled 37 patients in the studies. The follow-up period lasted at least 3 years (3-15 years) after the diagnosis had been established. In all cases a complete ophthalmological examination and actual ELISA test were performed. We compared the clinical status in two groups of patients: one with positive and the other with negative ELISA test at the time of control examination. RESULTS: In a majority of initially positive serological patients the control ELISA test for Toxocara canis antigen was negative. In these cases various post-inflammatory lesions in the anterior and posterior pole of the eye were present. In 8 cases the ELISA test was positive, despite the absence of active inflammatory process. In 5 serologically positive patients the active inflammation was observed. In more than 50% of cases the visual acuity was decreased. CONCLUSION: Ocular toxocariasis is a long lasting, severe type of uveitis that requires long treatment and causes dramatic visual impairment. ELISA test is a sensitive method indicating the intensity of inflammation in ocular toxocariasis. PMID- 9340421 TI - [Abducent nerve palsy as the only symptom of intracavernous carotid aneurysm in a child]. AB - A case of a 10-year old child with isolated sixth nerve palsy due to giant intracavernous carotid aneurysm is presented. The aneurysm was confirmed by CT brain scan and carotid angiography. It was closed after local embolization of internal carotid artery with the use of platinum inlays. It turns out that an intracranial aneurysm can be, although very rarely, the cause of isolated sixth nerve palsy in childhood. PMID- 9340422 TI - [Unilateral exophthalmus in the course of spontaneous retrobulbar hemorrhage in a patient with arterial hypertension]. AB - In this paper we present a case of a 60-year-old patient with labile hypertension, who was admitted to hospital because of unilateral exophthalmus and loss of vision of the left eye. During the hospitalization basic laboratory investigations, USG and NMR of orbits and arteriography of internal carotid arteries were carried out. Body temperature and arterial blood pressure were monitored. Leucocytosis and high values of arterial blood pressure in the early hours of the morning were found. Accessory investigations didn't explain the cause of exophthalmus. Repeated NMR of orbits showed image characteristic for retrobulbar haemorrhage in the left orbit. During the hospitalization hypotensive, anti-inflammatory and oedema-reducing treatment was administered, without surgical intervention. This case demonstrates that modern diagnostic investigations must be preceded by precise history taking concerning the early symptoms of the disease, its course and accompanying systemic diseases. Although spontaneous retrobulbar haemorrhages occur rarely, they must be taken into consideration in differential diagnosis of unilateral exophthalmus, especially in elderly patients with atherosclerosis and hypertension. PMID- 9340423 TI - [Selected problems with ocular accommodation in children and youth]. AB - This paper intends to present some accommodative disturbances which cause difficulties in reading and should be differentiated from dyslexia. The work describes the way of evaluation of accommodative convergence to accommodation rate (AC/A ratio). Clinical forms of two types of nonrefractive accommodative convergence excess connected with high AC/A ratio, namely hyperkinetic and hypoaccommodative, are presented. In hyperkinetic type, disturbances of ocular movements coordination during reading prevail. Patients with hypoaccommodation disorders suffer from youth presbyopia. The paper describes the ways of treatment of these disorders. PMID- 9340424 TI - [A new era in treatment of retinoblastoma]. PMID- 9340425 TI - [From the therapy study to quality assurance in pediatric oncology]. PMID- 9340426 TI - [Detection of translocation t(8;14)(q24;132) in pediatric Burkitt's lymphomas using "long distance" polymerase chain reaction: a new method for diagnosis of Burkitt's lymphomas]. AB - The pediatric Non-Hodgkin's lymphomas are a heterogeneous group of malignancies of B- or T-cell origin. Approximately half of them are characterized as Burkitt's lymphomas. Typically, one of the reciprocal translocations t(8;14)(q24;q32), t(2;8)(p11;q24) or t(8;22)(q24;q11) is seen in the tumor cell, each involving the protooncogene c-myc on chromosome 8. Characteristically, in most patients the translocation occurs between the distal end of the long arm on chromosome 8 (c myc) and chromosome 14 (immunoglobulin heavy chain locus, IgH). The breakpoint regions are distributed over a wide range of more than 10 Kb on chr. 8 and over several hundred Kb on chr. 14. With standard-PCR, fragments can only be amplified to a size of about 2 Kb. The development of PCR-applications to generate long products up to 20 Kb now allows a detection of these breakpoints. Several primer pairs from different regions of the IgH-gene and the c-myc-gene were tested in each patient. Until now, 20 patients with Burkitt's Lymphoma or B-ALL characterized by L3 morphology were examined. All patients were treated according the protocols of the NHL-BFM '90 or '95 study. In 11/20 patients, recombinations between chromosomes 8 and 14 could be detected with our primer pairs. In serial dilutions of DNA from malignant cells in DNA from healthy controls, sensitivities of one malignant cell in 2 x 10(4) normal cells could be obtained. This method will now allow us to characterize the involved breakpoints more exactly and to analyze patient samples (blood, bone marrow, aphereses products and residual tumors) during or after therapy for the existence of minimal residual tumor cells. PMID- 9340428 TI - [Kinetics of myelopoietic regeneration and mobilization of CD34-positive cells within the scope of the NB90 Neuroblastoma Therapy Study]. AB - Hematological and clinical data of 14 children with neuroblastoma treated according to the German neuroblastoma therapy study NB 90 were analyzed. Therapy included 4 or 8 intensive therapy elements N1 (Etoposide 125 mg/m2 day 1-4, Vindesine 3 mg/m2 day 1, Cisplatin 40 mg/m2 day 1-4) and N2 (Vincristine 1.5 mg/m2 day 1 + 8, Dacarbazine 200 mg/m2 day 1-5, Ifosfamide 1500 mg/ m2 day 1-5, Doxorubicin 30 mg/m2 day 6 + 7) in alternating order. The hematological recovery was studied after 86 therapy elements N1/N2. G-CSF had been given in 23 therapy courses, while no cytokine was administered in 63 therapy courses. Mobilization of CD34+ cells was studied in 13 therapy courses with G-CSF. Severe myelosuppression with an absolute neutrophil count < 500/microL was noted 2-4 weeks after each therapy element. The use of G-CSF did not prevent, but shortened neutropenia. There was no difference in the number of infections nor time delay of therapy between the courses with or without G-CSF. In 11 therapy courses G-CSF was started on the day following the last chemotherapy dose (N1: day 5; N2: day 9). In 12 therapy courses G-CSF was given delayed, starting day 12 after the initiation of therapy. Kinetics of granulocyte recovery was similar in the early or delayed application of G-CSF. Neutrophil recovery after the therapy element N1 was earlier and faster compared to that of therapy element N2. The more rapid rise of the neutrophils after the N1 element was accompanied by an effective mobilization of CD34+ cells. Taking into account the limitations of this retrospective study, the data may help to optimize the application of G-CSF in a very intensive therapy study like NB90. PMID- 9340427 TI - [Beta thalassemia in Germany: molecular genetics and clinical phenotype in immigrant and in the native population]. AB - BACKGROUND: In Germany there are about 300-400 patients with homozygous beta thalassaemia who immigrated from endemic regions mostly in the Mediterranean. In the non-immigrant population beta-thalassaemia is rare with only single case reports of homozygous patients. Heterozygous beta-thalassaemia, however, is more common and must be considered in the differential diagnosis of hypochromic anemia. PATIENTS AND METHODS: Here, clinical and molecular data of 221 homozygous patients from immigrant families and 256 non-immigrant German heterozygous individuals are presented. RESULTS: Clinically, 87% (n = 192) of the homozygotes are regularly transfused and classified as thalassaemia major (TM). The other 13% (n = 29) are not (regularly) transfused and thus classified as thalassaemia intermedia (TI). There is a wide spectrum of 39 beta-globin gene mutations and even the most common three, IVSI-110G-A, NS 39, and IVSI-6 T-C occur with relatively low frequencies of 28%, 22%, and 9%, respectively. In 17/29 (58%) TI patients "mild" mutations are found that inactivate the affected gene incompletely. In 16/29 (55%) there are mutations that are associated with increased gamma-globin gene activity. alpha-Thalassaemia is rare and only found in 3/29 TI-patients. In the 256 Germans with heterozygous beta-thalassaemia there are 27 different beta-globin gene mutations. The 3 most common are Mediterranean mutations together accounting for 61%. Also-relatively common (5%; n = 13) is an otherwise rare frameshift mutation of codon 83 (FS83 delta G). The other mutations occur in < 10 individuals only. Two mutations described here are novel. One of them affects position-2 of the intron 1 splice acceptor site (IVSI-129 A G) and the other is a deletion of a single G in codon 15/16 (FS 15/16 delta G). CONCLUSIONS: Taken together, a plausible molecular pathogenesis for the observed phenotype (TM vs. TI) can be identified in most homozygous patients thus allowing for rational counselling of the affected families. In heterozygous Germans beta thalassaemia has probably been imported from the Mediterranean in about 2/3 of the cases whereas in the remaining 1/3 it has probably originated locally. PMID- 9340429 TI - [Structural and functional changes in the heart in dermatomyositis and polymyositis]. PMID- 9340430 TI - [Myasthenic cardiomyopathy]. PMID- 9340431 TI - [Clinical implications of high blood viscosity and its reduction bu photo hemotherapy]. AB - The paper is concerned with an essential blood characteristic--viscosity which is considered in relation of mechanisms responsible for its normal and abnormal characteristics, its role in the onset of circulatory disorders. High blood viscosity may contribute to development of vascular disturbances. Efficient methods of normalizing high blood viscosity are analyzed with special emphasis on photochemotherapy. Effects of ultraviolet, blue and red laser radiation on blood viscosity are outlined. PMID- 9340432 TI - [C-reactive protein in SLE males: contribution to thrombotic complications]. PMID- 9340433 TI - [Surgical treatment of postinfarction left ventricular aneurysm]. PMID- 9340434 TI - [Effectiveness of drug therapy alone or its combination with plasmapheresis in patients with severe angina pectoris]. AB - 92 patients with ischemic heart disease and severe angina pectoris received either nitro drugs and calcium antagonists (n = 24) or their combination with plasmapheresis (n = 68). The efficacy of the treatment was assessed by the number of anginal episodes, doses of nitro drugs, pre- and posttreatment dynamic ECG readings. It is shown that 75% of the patients benefited from the addition of plasmapheresis to the treatment scheme. According to dynamic ECG, positive effect of plasmapheresis occurred also in painless ischemia. PMID- 9340435 TI - [New proofs of active oxygen forms involvement in pathogenesis of bronchial asthma]. PMID- 9340436 TI - [Comparison of myocardial effects of various antirheumatic modalities in patients with rheumatoid arthritis (prospective clinico-echocardiographic study)]. AB - A prospective study was conducted of basic antirheumatic drugs (chrisanolum, myocrisin, ridaura, chlorbutin, D-penicillamine) versus nonspecific antiinflammatory treatment in 127 patients with rheumatoid arthritis (RA). The drugs were compared by their effect on myocarditis. Basic antirheumatic drugs were found much superior to nonsteroid antiinflammatory drugs. The highest response was registered for chrisanolum myocrisin and chlorbutin after 6-12 month continuous treatment. Warning: chrisanolum, myocrisin and ridaura may produce unwanted short-term effects on left ventricular contractility in developing extracardiac complications of aurotherapy. PMID- 9340437 TI - [Hereditary diseases: principles of treatment]. PMID- 9340438 TI - [Sanatorium rehabilitation of cardiosurgical patients]. AB - The authors formulate a new approach to classification and therapy of clinical and mental complications after operations on the heart. Their clinical experience with 29% patients surgically treated for cardiac diseases (aortocoronary bypass and other operations) provided proofs of a good therapeutic effect of postoperative psychological correction performed in sanatorium (in the Central Army Sanatorium, in particular). PMID- 9340439 TI - [Combined diagnosis of primary lung cancer using immunological tests]. AB - To compare the accuracy of diagnosis obtained with different diagnostic techniques the authors examined 187 patients with cancer of the lung clinically, roentgenologically, bronchologically, morphologically and immunologically. X-ray made the diagnosis of lung cancer in 85% of the examinees. This diagnosis was confirmed in 87% of the cases. Morphological verification was obtained in 84.4% and 78.2% of patients with central and peripheral cancer, respectively. Additional immunological investigations increased the proportion of accurate diagnoses up to 96.8%. It is concluded that immunological investigations are effective in complex diagnosis of lung cancer. PMID- 9340440 TI - [Endoscopical methods in the treatment of postcholecystectomy syndrome patients]. PMID- 9340441 TI - [Comparative efficacy of single Berodual inhalation in patients with bronchial asthma: nebulization against dose-adjusted aerosol]. AB - The trial included 34 patients with moderate non-atopic asthma in exacerbation. After arrest of the acute attack the patients inhaled berodual via nebulizer (a single dose of 0.5 ml -125 micrograms ipratropium bromide and 250 micrograms phenoterol) or aerosol (40 micrograms ipratropium and 100 micrograms phenoterol). As shown by body plethysmography and peak-flowmetry, nebulizer therapy was characterized by earlier onset of broncholytic effect which stood longer, more pronounced effect at the level of small and middle bronchi, absence of side effects. The authors believe that berodual inhalations through nebulizer are an effective modality in the treatment of bronchial asthma in severe exacerbation. PMID- 9340442 TI - [Current potentialities of Buscopan in clinical practice]. PMID- 9340443 TI - [Permitted for use by Russian Federation Ministry of Health]. PMID- 9340444 TI - [Kidneys involvement in multiple myeloma. Therapeutic issues]. PMID- 9340445 TI - [Diagnosis of exudative pleurisy ]. PMID- 9340446 TI - [Long-standing subfebrile condition]. PMID- 9340447 TI - [Salmonellosis peritonitis]. PMID- 9340448 TI - [Gastrointestinal tuberculosis]. PMID- 9340449 TI - [Stolt vs Evans--a clinician meets a health economist. How much is the cost of dying?]. PMID- 9340450 TI - [No perforations with an intrauterine device approved by the EU]. PMID- 9340451 TI - [Are tricyclic antidepressants unsuitable in depression?]. PMID- 9340452 TI - [A name is not enough as identity code]. PMID- 9340453 TI - [Patient in the IT-age]. PMID- 9340454 TI - [Global spreading of resistant staphylococci. The best control provided by competent specialists at infection clinics]. PMID- 9340455 TI - [Quality assurance of asthma care within primary health care in the E-county. Fewer symptoms and emergency visits with a local care program]. PMID- 9340456 TI - [Therapeutic recommendations in asthma. The specialist reads them but not allways follows them]. PMID- 9340457 TI - [Fetal cardiology. A new specialty with great diagnostic and therapeutic possibilities]. PMID- 9340458 TI - [Social medicine has "let thousand flowers bloom". Public health science should be the basis]. PMID- 9340459 TI - [Local anesthesia is effective in Cesarean section]. PMID- 9340461 TI - [Control of the hand and its grip. Memory and touch perception are crucial factors]. PMID- 9340460 TI - [Gluten intolerance in children--diagnostic routines in Sweden 1996. Great variations in celiac disease studies]. PMID- 9340462 TI - [Methicillin resistant Staphylococcus aureus. A problem also in Sweden]. AB - Outbreaks of endemic MRSA are extremely rare in Swedish hospitals. During 1983 1996 209 patients with MRSA were identified at Sahlgrenska University Hospital, Goteborg. Retrospective DNA typing showed that 51 different strains were involved. Between 1990-1996 there was an increase in the number of imported MRSA infected patients, probably a reflection of the situation in surrounding high prevalence countries. In addition four endemic outbreaks occurred during the 1990s involving a total of 121 patients, mainly in units for intensive care and urology. For effective control of MRSA, standard precautions should include the use of plastic aprons and hand disinfectants (70 per cent alcohol) when nursing patients with wounds, urinary catheter etc regardless of MRSA-status. PMID- 9340463 TI - [A pause in the history of medicine]. PMID- 9340464 TI - [Cancer: quality assurance, prevention, new therapies]. PMID- 9340465 TI - [Cell adhesion molecules]. PMID- 9340466 TI - [New positive inotropic drugs]. PMID- 9340467 TI - [Hidden alcohol in foods?]. PMID- 9340468 TI - [Recommendations for implementing bronchial provocation tests with pharmacologic substances. German Society of Pneumology--Scientific "Bronchial Provocation Tests" Study Group]. PMID- 9340469 TI - [Antibiotic resistance of enterococci in Germany]. AB - BACKGROUND: The resistance of enterococci against various antimicrobial substances including vancomycin has increased markedly. Since 1989 in the USA in particular high resistance rates against vancomycin have been observed but very few surveillance have been published in Europe. Therefore, we conducted a multicenter study in Germany to obtain information about the incidence and distribution of vancomycin and/or high-level aminoglycoside-resistant enterococci. METHODS: A total of 2046 enterococcal isolates were identified and susceptibility-testing was performed according to international guidelines. RESULTS: A total number of 90.5% of the enterococcal isolates were identified as Enterococcus faecalis and 7.8% was Enterococcus faecium. Resistance against ampicillin was detected in 56.6% of the Enterococcus faecium isolates, however, in only one Enterococcus faecalis isolate. High-level resistance against gentamycin or streptomycin was observed in 7.3% and 24.8% of the isolates, respectively. Twelve isolates showed resistance against vancomycin, however, cross resistance with teicoplanin was found in only two isolates. CONCLUSION: The rate of resistance of enterococci in Germany is still considerably lower than in the United States. Previous vancomycin therapy has been implemented as a risk factor for colonization or infection with vancomycin-resistant enterococci. Continued vigilance, decreased use of vancomycin and strict enforcement of infection control measures are appropriate measures to control the growing problem of resistant enterococci. PMID- 9340470 TI - [Therapy of acute renal failure]. PMID- 9340471 TI - [Recurrent abdominal pain]. PMID- 9340472 TI - [Value of biguanide in therapy of diabetes mellitus]. AB - BACKGROUND AND OBJECTIVES: Biguanides have been used in treatment of diabetes mellitus for over 30 years now. Due to frequent occurrence of lactic acidosis, particularly in patients with serious contraindications to biguanide therapy and in cases of non-compliance with dosage instructions, buformin and phenformin were taken off the market in most European countries at the end of the seventies. Metformin continued to be allowed, since the risk of lactic acidosis is 20 times less than with phenformin or buformin due to the different pharmacokinetic properties of the substance. Plenty of clinical experience has been gained with metformin, documented in a large number of reliable long-term studies. FINDINGS: Metformin lowers fasting blood glucose levels by an average of 25% (17 to 37%), postprandial blood glucose by up to 44.5% and HbA1c bei 1.5% (0.8 to 3.1%) Metformin reduces raised plasma insulin levels in cases of metabolic syndrome by as much as 30% and reduces the "insulin requirement" of type 2 insulin-treated diabetics by 15 to 32%. It has well documented effects on various rheological parameters. In overweight type 2 diabetics, metformin shows the same level of hypoglycaemic effect as all of the important sulfonylurea derivatives used in Europe. The active mechanism of these derivatives is, however, concentrated solely on reduction of blood glucose. This mechanism does not take into account the remaining risk constellation involved in insulin resistance. Biguanides, similarly to weight reduction, lead to a reduction of hyperinsulinaemia, which is by contrast exacerbated by sulfonylureas and, in particular, exogenous insulin. CONCLUSION: The risk of lactic acidosis can probably be eliminated entirely if dosage instructions and contraindications are observed carefully. The cause of such neglect in 83% of all cases was limited on renal function (serum creatinine > 1.5 mg%). Regarding morbidity and mortality from lactic acidosis, metformin therapy is no riskier than treatment with the sulfonylurea derivative glibenclamide, taking into account the incidence of fatal hypoglycaemias with the latter. PMID- 9340473 TI - [Transplantation of hematopoietic stem cells. I: Definitions, principle indications, complications]. AB - The transplantation of hematopoietic and lymphopoetic stem and progenitor cells has become a standard procedure for the treatment of many malignant diseases. Autologous stem cells are derived from the patient himself, allogeneic cells from an HLA-identical or HLA-compatible family or unrelated donor. Hematopoietic stem cells can be obtained from bone marrow, blood and fetal cord blood. After 3 to 5 days treatment, the granulocyte-colony stimulating factor (G-CSF) mobilizes stem- and progenitor cells from the marrow into the blood. This method is now standard in autologous transplantation and is increasingly preferred in allogeneic transplantation. The time to hematopoietic recovery is shorter with blood stem cells than with bone marrow cells. With myeloablative high dose therapy followed by stem cell transplantation, long term disease free survival is possible in many cases and great proportions of patients can be cured (see part II). Improvements of supportive care have reduced toxicity of treatment substantially, however severe complications still occur at oropharynx, gastrointestinal tract, liver, lung, skin, kidney, urinary tract and nervous system. After allogeneic transplantation immunocompetent donor cells can react with the recipients tissue. In HLA-identical donor and recipients differences in the minor histocompatibility antigens account for this graft-versus-host-reaction (GvH), which is mainly mediated by transplanted T-cells. The GvH-reaction can affect skin, liver, gut and other organs and cause clinically relevant GvH-disease (GvHD). The GvHD is more severe in HLA-mismatched or unrelated transplantations. Immunodeficiency and organ dysfunction due to GvHD may predispose infections and impair the outcome of transplantation. Unrelated cord blood stem cells may have a minor risk of inducing acute GvHD, as stem and T-cells are immature. After allogeneic stem cell transplantation, the relapse rate of leukemia or lymphoma is significantly reduced by immunoreactive cells:graft-versus-tumor (GvT) or graft-versus-leukemia effect (GvL). PMID- 9340474 TI - [Is the bronchial provocation test a provocation?]. PMID- 9340475 TI - [Hyperferritinemia in Still syndrome in the adult and reactive hemophagocytic syndrome]. AB - This report describes the fatal outcome of a case of adult onset Still's disease in a 46-year old man. The diagnosis was made according to the 1992 criteria, proposed by Yamaguchi. Nine months after the initial disease manifestations a rapid deterioration with progressive hepatosplenomegaly developed. In parallel, pancytopenia and marked hyperferritinemia could be detected. Transjugular liver biopsy revealed the presence of a hemophagocytic syndrome. The course of the disease was refractory to any form of treatment and the patient died from disseminated intravascular coagulation, hepatic and pulmonary failure. Pathogenetic mechanisms and possible associations between Still's disease and reactive hemophagocytic syndrome are discussed. PMID- 9340477 TI - [News in virology for pediatricians]. PMID- 9340476 TI - [Cost effectiveness of bisoprolol in heart failure. Economic evaluation of the Cardiac Insufficiency Bisoprolol Study (CIBIS) for Germany]. AB - BACKGROUND: The Cardiac Insufficiency Bisoprolol Study (CIBIS) demonstrates that, for patients with heart failure of different etiologies, the administration of the beta(1)-adrenoceptor blocker bisoprolol adjuvant to the standard therapy leads to a significant avoidance of hospital admissions. PHARMACOECONOMIC EVALUATION: The results of the CIBIS were evaluated pharmacoeconomically for the Federal Republic of Germany, and were restricted to direct costs only. The costs of bisoprolol medication and in-patient treatment of heart failure were considered, the latter forming the major part of costs incurred. CONCLUSION: Adjunctive therapy with bisoprolol is not only clinically beneficial to the patient with heart failure, but also economically advantageous. PMID- 9340478 TI - [Trend of circulating eosinophils in healthy children and children suffering from infectious diseases. A retrospective study]. AB - INTRODUCTION: Acute bacterial and viral infections are accompanied by a marked diminution in circulating eosinophils in the blood. This forms part of the host's physiological response to acute infection and was first studied in adults early this century. The aims of this study were to check whether eosinopenia during acute phlogosis is a phenomenon present in pediatric patients, and whether the trend is comparable to the experimental models reported; to describe the trend of circulating eosinophils in the remission process. METHODS: A retrospective study was performed in 34 children hospitalised in the Pediatric Hospital of AUSL 2 Lucca (Italy) for bacterial or viral infective diseases documented by cell culture or presumed diagnosed. Children with the following characteristics were excluded from the study: 1) blood samples collected for hemochrome analysis at times other than normal (7-8 a.m.); 2) cortisone treatment administered up to 5 days prior to blood sample and/or during hospitalisation; 3) positive personal anamnesis for manifest allergic diseases. On admittance (children during acute phase) and at the start of remission, an absolute count of circulating eosinophils was performed in these children using an automatic globule counter. Sixty-six children with non evident infective and/or inflammatory diseases were included in the study as a control group. This group was also selected in the same way as infective subjects. RESULTS: The mean number of circulating eosinophils was 288 (+/- 248) in the control group, 46 (+/- 58) in subjects at the acute phase of infective pathology and 252 (+/- 162) in infective patient during the remission phase. The difference between the two means was statistically significant. This characteristic falling and rising trend of circulating eosinophils was found in 33 of the 34 infective subjects examined. CONCLUSIONS: Eosinophil values found in control subjects are broadly in line with those reported in the literature. Eosinopenia during the course of acute infection and the early rise during remission represent a characteristic phenomenon indicating the body's "normal" response to a non-parasitic infection. Both eosinophil levels, namely in the control group (288/mm3) and in acute-phase subjects (46/mm3), should be regarded as "normal" provided they refer to the appropriate situation. The precocity and precision with which the eosinophil trend follows the phases of the infection underlines the value of the assay of these cells as a reliable parameter for monitoring acute infection. There are also indications that, in an inflammatory situation, the behaviour of circulating eosinophils may provide a practical clinical marker of the predominant lymphocyte pattern (Th1 or Th2), as well as the phase of phlogosis, active or remission. PMID- 9340479 TI - [Pattern of circulating eosinophils in allergic children suffering from infectious disease]. AB - BACKGROUND AND AIM: Both bacterial and viral acute infections are accompanied by a typical fall in the number of circulating eosinophils followed, at remission, by a rapid return to starting values. The aim of this study was to check whether this typical pattern can be observed in atopical children. MATERIALS AND METHODS: A retrospective study was performed in a group of atopical subjects (mainly asthmatic) suffering from intercurrent infectious diseases (mainly bronchopneumonia); circulating eosinophils were assayed during the acute phase of disease and during remission. The control group consisted of 21 atopic children without signs or symptoms of infectious diseases. RESULTS: The mean number of circulating eosinophils was 412.6 +/- 199.8 in the control group of allergic children 25.1 +/- 39.7 in the group with acute infectious diseases and 241 +/- 137.5 in the group of children with infectious diseases during remission. The differences between the group of children with infectious diseases and that in remission were statistically significant. The typical pattern of the fall and rise of circulating eosinophils was observed in 9 out of 10 children with acute infections. The cut-off proposed as the discriminating factor between acute phase infection and subjects in remission or without infectious diseases was 100 eosinophils/mm3. CONCLUSIONS: During the course of infective pathologies in allergic subjects there is an abrupt and significant reduction in circulating eosinophils followed, during remission, by a return to levels comparable to those in controls. Eosinophil assay therefore appears to be a useful parameter to monitor acute infection in allergic subjects. In particular, increased eosinophil levels are an early indicator that the infection has been overcome and remission is in progress. The authors repropose the hypothesis that eosinophils play a double role that alternates between activating the allergic phlogosis and eliminating non-allergic phlogosis, depending on the cytokinic context in which this occurs. PMID- 9340480 TI - [Conservative treatment of dysplastic multicystic kidney]. AB - BACKGROUND: Recent developments regarding the conservative management of multicystic dysplastic kidney disease (MDKD) have shown a reduction in the well established practice of elective surgical treatment. It has been seen that in the majority of cases the evolution of MDKD is either partial or complete atrophy. The incidence of complications due to MDKD is very low. The aim was to study the ultrasound changes of MDKD in a group of patients from 1990, since this date the elective surgical procedure had been eliminated. MATERIALS AND METHODS: This group consisted of 16 patients. The diagnosis in each patient was confirmed by ultrasound and isotopic examinations. Other investigations were performed where there was a suspect of associated contralateral renal abnormalities or when a urinary tract infection occurred. RESULTS: Ultrasound follow-up revealed: no enlargement whatsoever in the kidney size in any patient; in 10 patients a reduction in the size of the MDK; in 3 patients a complete atrophy of MDK; in 3 patients there was no change at all. Five patients were found to have contralateral renal abnormalities. Compensatory hypertrophy of the contralateral kidney was present in all patients. No complications occurred. A nephrectomy was performed only in one patient aged 6 due to the express wishes of his parents. CONCLUSIONS: The results agree with recently published literature that routine surgical removal of the MDK is unnecessary. PMID- 9340481 TI - [Nailfold capillaroscopy in normal children between 0 and 16 years of age]. AB - BACKGROUND: Nailfold capillaroscopy is a technique used in childhood and adults for morpho-functional study of peripheral microcirculation. The aim of the present study is to evaluate the evolution of the microcirculation. METHODS: Nailfold capillaroscopy was performed in eighty-six healthy children (48 boys and 38 girls) aged 5 months to 16 years. All the subjects were divided into 5 groups for age. RESULTS: A progressive morphological and dynamic development of peripheral microcirculation in the early years of life was observed. CONCLUSIONS: Therefore, nailfold capillaroscopy is a simple, noninvasive, easily repetitive method which allows to observe the physiological development and a possible pathology of naifold capillaries. PMID- 9340482 TI - [Alcohol consumption in schoolchildren. Results of a questionnaire study]. AB - BACKGROUND AND METHODS: Alcohol is the leading cause of acute poisoning in Trieste teenagers and one of the main public health problems in general population. Planning of effective prevention is limited by lack of information on alcohol drinking habits and notions of young people. MATERIALS AND METHODS: A questionnaire study has been carried out on 769 school teen-agers in Trieste and Gorizia provinces. The anonymous questionnaire concerned alcohol use and daily consumption, age and site of first approach, favourite beverages, number of acute abuse during the last year, awareness of the problem and opinion on social aspects. RESULTS: Near half of school teen-agers reported alcohol drinking. First approach occurred more frequently at home, because of curiosity or parents emulation and it was especially precocious for wine and beer (mode 8-10 yrs) respect to spirits (mode 12 yrs). In males and in both gender with increasing age higher rates of drinkers, alcohol consumption and acute poisoning were reported. More than 8 of 10 children judged the alcohol to be not a food but a drug, whose consumption is not necessary to affirm the personality. However, for more that 8 of 10 subjects alcohol warms, for about half of them it is stimulant. Only 4 of 10 were aware that its sale is forbidden by law under 16 years of age. CONCLUSIONS: Due to early initiation, family responsibility and lack of awareness of the problem, a correct alcohologic information to families and teen-agers is needed. Pediatricians can and must have a main role at this regard, but their action must be associated with reduction of our society tolerance on alcohol drinking. PMID- 9340483 TI - [Orbital cellulitis. A case report]. AB - This study describes the case of a 6 years old child, male, with orbital cellulitis and underlines the importance of an early diagnosis and therapy to avoid severe complications often present in this disease. Swelling and redness of the eyelid, pain and ophthalmoplegia are the first sign of an orbital cellulitis and they require rapid diagnostic procedure such as ultrasound and TC scan of the orbital region to evaluate the integrity of the profound orbital tissues. The child was admitted at the Department of Pediatrics, University "La Sapienza" of Rome and underwent an ultrasound, TC scan and serum exams which demonstrated the elevation of the sedimentation rate, reactive C protein and WBC plus the interesting of the profound orbital tissues. The child was treated with antibiotic and antiinflammatory therapy showing a complete recovery within 7 days. An ultrasound performed 7 days later demonstrated a complete resolution of the inflammatory process. In summary, this study would like to stress the necessity of an early diagnosis and an appropriate therapy in order to avoid the severe complications often present in children with orbital cellulitis. PMID- 9340484 TI - [Ehlers-Danlos syndrome type I. Ultrastructural study]. AB - Ehlers-Danlos syndrome comprises a very heterogeneous group of collagen diseases characterised in clinical terms by fragility and cutaneous hyperextensability, ligamentous hyperlaxity, ecchymosis, scarring, visceral and neurological manifestations. Having been described in detail by Ehlers in 1899 and Danlos in 1908, it was subsequently classified into various clinical types. At present at least 11 forms are recognised on the basis of their clinical characteristics, methods of transmission and biochemical defect; the first four types of the syndrome account for approximately 95% of cases. Almost all forms are transmitted with a dominant autosomic character. Specific genetic mutations have been ascertained whereas the biochemical defect has been identified in numerous types. Ehlers-Danlos type 1 syndrome is the most frequent and most severe form. The biochemical anomaly underlying the altered deposition of collagen fibre is still unknown and this is responsible for the "storiform" appearance of collagen fibre on ultrastructural examination. The authors have described a typical case of "Ehlers-Danlos type 1 syndrome" in which the diagnosis was confirmed by comparing clinical data and the results of ultrastructural tests which revealed the characteristic "pattern" of collagen fibres. PMID- 9340485 TI - [Determination of the gestational age]. PMID- 9340486 TI - [Transcriptional regulation of eukaryotic genes: data bases and computer analysis]. PMID- 9340487 TI - [Classification of eukaryotic transcription factors]. PMID- 9340489 TI - [Description of eukaryotic promoters in the EPD database]. PMID- 9340488 TI - [Composite regulatory elements: classification and description in the COMPEL database]. PMID- 9340490 TI - [TRRD: a database of transcription regulatory regions in eukaryotic genes]. PMID- 9340491 TI - [Integrating knowledge on transcriptional regulation of eukaryotic genes based on information from TRANSFAC, TRRD, and COMPEL databases]. PMID- 9340492 TI - [The MGL computer system--an instrument for generating, graphically representing, and analyzing regulatory genomic sequences]. PMID- 9340493 TI - [Intergenic interrelations in regulating the cell cycle: key role of E2F family transcription factors]. PMID- 9340494 TI - [Mechanisms of transcriptional regulation of erythroid specific genes]. PMID- 9340495 TI - [Transcriptional regulation of lipid metabolism genes: description in the TRDD database]. PMID- 9340496 TI - [Mechanisms of transcriptional regulation of interferon-induced genes: description in the IIG-TRRD information system]. PMID- 9340497 TI - [Mechanisms of glucocorticoid regulation and regulatory regions of genes, controlled by glucocorticoids: description in the TRDD database]. PMID- 9340498 TI - [Modeling TATA-box sequences in eukaryotic genes]. PMID- 9340499 TI - [Computer analysis of conformational features of the eukaryotic TATA-box DNA promotors]. PMID- 9340500 TI - [Detection of correlating DNA-binding positions in CREB and AP-1 family transcription factors]. PMID- 9340501 TI - [Analysis of transcriptional regulatory regions based on detecting transcription factor binding sites and studies of their mutual position]. PMID- 9340502 TI - [Genetic level of DNA sequences is determined by superposition of many codes]. PMID- 9340503 TI - A comparison of reteplase with alteplase for acute myocardial infarction. AB - BACKGROUND: Reteplase (recombinant plasminogen activator), a mutant of alteplase tissue plasminogen activator, has a longer half-life than its parent molecule and produced superior angiographic results in pilot studies of acute myocardial infarction. In this large clinical trial, we compared the efficacy and safety of these two thrombolytic agents. METHODS: A total of 15,059 patients from 807 hospitals in 20 countries who presented within 6 hours after the onset of symptoms with ST-segment elevation or bundle-branch block were randomly assigned in a 2:1 ratio to receive reteplase, in two bolus doses or 10 MU each given 30 minutes apart, or an accelerated infusion of alteplase, up to 100 mg infused over a period of 90 minutes. The primary hypothesis was that mortality at 30 days would be significantly lower with reteplase. RESULTS: The mortality rate at 30 days was 7.47 percent for reteplase and 7.24 percent for alteplase (adjusted P=0.54; odds ratio, 1.03; 95 percent confidence interval, 0.91 to 1.18). The 95 percent confidence interval for the absolute difference in mortality rates was 1.1 to 0.66 percent. Stroke occurred in 1.64 percent of patients treated with reteplase and in 1.79 percent of those treated with alteplase (P= 0.50). The respective rates of the combined end point of death or nonfatal, disabling stroke were 7.89 percent and 7.91 percent (P=0.97; odds ratio, 1.0; 95 percent confidence interval, 0.88 to 1.13). CONCLUSIONS: As compared with an accelerated infusion of alteplase, reteplase, although easier to administer, did not provide any additional survival benefit in the treatment of acute myocardial infarction. Other results, particularly for the combined end point of death or nonfatal, disabling stroke, were remarkably similar for the two plasminogen activators. PMID- 9340505 TI - Primary coronary angioplasty versus thrombolysis. PMID- 9340506 TI - Primary coronary angioplasty versus thrombolysis. PMID- 9340507 TI - Cyclosporiasis and raspberries. PMID- 9340504 TI - A comparison of continuous infusion of alteplase with double-bolus administration for acute myocardial infarction. AB - BACKGROUND: Accelerated infusion of alteplase (tissue plasminogen activator) over a period of 90 minutes induces more rapid lysis of coronary-artery thrombi than a 3-hour infusion. With two bolus doses of alteplase, further shortening the duration of administration, complete reperfusion was achieved in more than 85 percent of the patients in initial angiographic studies. We tested the hypothesis that double-bolus alteplase is at least as effective as accelerated infusion. METHODS: In 398 hospitals, 7169 patients with acute myocardial infarction were randomly assigned to weight-adjusted, accelerated infusion of 100 mg of alteplase or to a bolus of 50 mg of alteplase over a period of 1 to 3 minutes followed 30 minutes later by a second bolus of 50 mg (or 40 mg for patients who weighed less than 60 kg). The primary end point was death from any cause at 30 days. The trial was stopped prematurely because of concern about the safety of the double-bolus injection. RESULTS: Thirty-day mortality was higher in the double-bolus group than in the accelerated-infusion group: 7.98 percent as compared with 7.53 percent. The absolute difference was 0.44 percent, with a one-sided 95 percent upper boundary of 1.49 percent, which exceeded the prespecified upper limit of 0.40 percent to indicate equivalence in 30-day mortality between the two regimens. The respective rates of any stroke and of hemorrhagic stroke were 1.92 and 1.12 percent after double-bolus alteplase, as compared with 1.53 and 0.81 percent after an accelerated infusion of alteplase (P=0.24 and P=0.23, respectively). CONCLUSIONS: Double-bolus alteplase was not shown to be equivalent, according to the prespecified criteria, to accelerated infusion with regard to 30-day mortality. There was also a slightly higher rate of intracranial hemorrhage with the double-bolus method. Therefore, accelerated infusion of alteplase over a period of 90 minutes remains the preferred regimen. PMID- 9340508 TI - Cyclosporiasis and raspberries. PMID- 9340509 TI - Cyclosporiasis and raspberries. PMID- 9340510 TI - Cyclosporiasis and raspberries. PMID- 9340511 TI - Chemotherapy for AIDS-related lymphomas. PMID- 9340512 TI - Chemotherapy for AIDS-related lymphomas. PMID- 9340513 TI - Solitary extramedullary plasmacytoma. PMID- 9340514 TI - Obstructive uropathy secondary to endometriosis. PMID- 9340515 TI - Duplicate publication and correction. PMID- 9340516 TI - Birth defects among children of Persian Gulf War veterans. PMID- 9340518 TI - Should legislatures practice medicine? PMID- 9340517 TI - The Supreme Court and physician-assisted suicide--rejecting assisted suicide but embracing euthanasia. PMID- 9340520 TI - Should legislatures practice medicine? PMID- 9340519 TI - Should legislatures practice medicine? PMID- 9340521 TI - Should legislatures practice medicine? PMID- 9340522 TI - Should legislatures practice medicine? PMID- 9340523 TI - Corticosteroid injections for sciatica. PMID- 9340524 TI - Corticosteroid injections for sciatica. PMID- 9340525 TI - Corticosteroid injections for sciatica. PMID- 9340526 TI - Corticosteroid injections for sciatica. PMID- 9340527 TI - Platelet glycoprotein IIb/IIIa receptor blockade after coronary angioplasty. PMID- 9340528 TI - Platelet glycoprotein IIb/IIIa receptor blockade after coronary angioplasty. PMID- 9340529 TI - Ticlopidine and thrombotic thrombocytopenic purpura. PMID- 9340530 TI - DNA testing to refute a diagnosis of cancer. PMID- 9340531 TI - Insurance copayments and delays in seeking emergency care. PMID- 9340532 TI - Insurance copayments and delays in seeking emergency care. PMID- 9340533 TI - Black widow spider. PMID- 9340534 TI - The compromise of compassionate care. North Carolina poet's essay illuminates doctors' role in caring for the terminally ill. PMID- 9340535 TI - [Deliberation between physician and patient concerning active euthanasia at the patient's home]. AB - Three patients, one man aged 68 and two aged 67, with terminal incurable cancer, requested euthanasia. It was performed in two, the third patient eventually died without having repeated his request. There are three phases in euthanasia: orientation (the patient asks the physician whether he would be willing to assist should the need arise), organisation (the physician ensures that the necessary prerequisites are fulfilled, i.e. the patient's request must be voluntary, mature and longlasting, his suffering must be longlasting, unbearable and incurable, and another physician must have been consulted and must have prepared a written report), and the phase entered after the definitive decision to perform euthanasia has been taken. The physician should not be reluctant to bring up the subject at an early stage, as it may set the patient's mind at rest to have expressed a wish concerning suffering and the end of life. PMID- 9340536 TI - [Taking the temperature]. AB - Fever is still an important indicator of disease, and an indication for diagnosis and treatment. Accurate measurement of the temperature is therefore desirable. The infrared tympanic thermometer abolishes the necessity of axillary, oral and rectal measurement, which are less reliable, and unpleasant to the patient. PMID- 9340537 TI - [Congenital long QT syndrome]. AB - The long QT syndrome (LQTS) combines a prolonged QT interval with an enhanced risk of polymorphous ventricular arrhythmias that may lead to syncope and sudden cardiac death. It may be congenital or acquired (the latter sometimes caused by drugs). Congenital LQTS is a rare disease, usually discovered during the clinical evaluation of understood syncopes or at cardiological examination after an unexpected sudden cardiac death of a close relative. The syncope frequently occurs during physical exercise, fear or sudden loud noises. In patients with symptomatic LQTS, the mortality 10 years after the first syncope amounts to approximately 50%. A prolonged QT interval indicates abnormal repolarization or deceleration of the depolarization. An increase of the sympathetic tone, e.g. during physical exercise and emotions, causes prolongation of the QT interval. Congenital LQTS has been associated with genetic mutations, for instance on chromosomes 3 and 7. Treatment consists af administration of beta-blockers, sympathectomy and, if necessary, implantation of an automatic cardioverter/defibrillator. PMID- 9340538 TI - [Clinical thinking and decision making in practice. A patient with flank pain and anuria]. AB - A 49-year-old man was admitted because of acute anuria. He had bilateral loin pain and was said to have been suffering from renal calculi seven years earlier. He used diclofenac as a pain killer. The renal pyela were mildly dilated. Eventually it became clear that the patient suffered from retroperitoneal fibrosis. This case illustrates (a) ureteral patency does not ensure actual urine flow; (b) non-steroidal anti-inflammatory drugs (NSAIDs) are a frequent cause of renal function loss, but only if there is an additional renal disease causing increased renal prostaglandin synthesis; (c) pattern recognition and verification are clinically relevant diagnostic tools. PMID- 9340539 TI - [Reading children's temperatures with the tympanic infrared thermometer and the rectal mercury thermometer: equally good results in the emergency room]. AB - OBJECTIVE: To compare the results of reading body temperatures with a tympanic infrared thermometer and a rectal mercury thermometer in children in an emergency department. DESIGN: Prospective comparative study. SETTING: St. Elisabeth Hospital, Tilburg, the Netherlands. METHOD: In children up to 11 years of age seen in the emergency room between 1 January 1994 and 1 April 1994, the body temperature was measured with a rectal mercury thermometer as well as with a tympanic infrared thermometer. Data were collected on temperature read, clinical picture on arrival (not ill, ill, seriously ill) and appearance of the tympanic membrane (signs of acute otitis media, presence of cerumen). For the statistical comparison, the differences between the findings of the two methods were plotted against the means. The sensitivity and specificity of the results of tympanic measurement in relation to the values read rectally were determined. RESULTS: Data were collected on 213 children, of whom 19 were younger than 3 months, 46 between 3 and 12 months, and 148 between 1 and 11 years. The mean temperatures measured with the rectal and tympanic thermometers were 38.01 and 38.03 degrees C, respectively. The mean difference between the rectal and tympanic temperatures was -0.013 degree C. The correlation between the rectal and tympanic temperatures was high (r = 0.86; P = 0.0001). The results were the same in groups differing in age, severity of disease and appearance of the tympanic membrane. The sensitivity of the tympanic measurement for fever (rectal temperature > 38.0 degrees C) was 80.6% with a specificity of 92.5%. The sensitivity was 83.8% when a rectal temperature > 38.5 degrees C was taken as the criterion, with a specificity of 95.9%. CONCLUSION: The tympanic infrared measurement in children in an emergency department gave the same results as rectal measurement using a mercury thermometer. PMID- 9340540 TI - [Measurement of the body temperature of adults by rectal digital thermometer and the infrared tympanic thermometer: equally good results in the department of internal medicine]. AB - OBJECTIVE: To examine whether, in clinical practice, the infrared tympanic thermometer shows temperature readings similar to those obtained with the rectal digital thermometer, so that the former can replace the latter. DESIGN: Prospective comparative study. SETTING: Academic Hospital, Nijmegen, the Netherlands. METHOD: In 104 patients admitted to the department of medicine, body temperature was measured by both methods within approximately ten minutes. This was done on two successive days. The measurements were then analysed by plotting the difference between two measurements against their mean. Then the limits of agreement, which are the mean of the differences between the two measurements plus and minus 2 standard deviations, were determined. With both thermometers also duplicate measurements were made to study the repeatability. RESULTS: The mean difference between the 2 methods in the first measurement was 0.15 degree C (SD: 0.56), in the second measurement it was 0.07 degree C (0.52). The limits of agreement were 1.27 degrees C and -0.97 degree C for the first comparisons and 1.13 degrees C and -0.99 degree C for the second comparisons. In the duplicate measurements, the mean difference between the first and the second measurement was 0.02 degree C (0.19) in the rectal measurement, and 0.09 degree C (0.23) in the tympanic measurement. The patients found the tympanic measurements significantly less painful and unpleasant than the rectal measurement. The mean time needed for the tympanic measurements (8 s) was ten times less than for the rectal measurements (79 s). CONCLUSION: The results of this study show good agreement between the infrared tympanic thermometer and the rectal digital thermometer so that they may be regarded as interchangeable. The patients had a clear preference for the tympanic thermometer, which also took less time. PMID- 9340541 TI - [The role of the consulting physician in situations of active euthanasia]. AB - The cases are reported of two patients, a man aged 69 with a metastasized bronchial carcinoma and a woman aged 65 with a frontotemporal glioblastoma no longer responding to irradiation. Both requested active euthanasia. In both cases, euthanasia was performed by injection, after a general practitioner from the same locum group had acted as consultant. The requirements of meticulousness in handling a request for active euthanasia are concerned with the request (which has to be voluntary, thoroughly considered and constant), the suffering (which has to be protracted, unbearable and incurable), consultation and the written report. The consulting or second physician in cases of active euthanasia confirms that the requirements of meticulousness have been met. In addition, the second physician may assist the general practitioner in the detection of factors that may impair correct decision-making by the doctor or the patient. The second physician will be aided in performing these tasks if he is a member of the same locum group as the treating physician. However, if he considers himself too involved, a physician outside the locum group should be available at all times. PMID- 9340543 TI - [Clinical thermometry. I. Historical developments]. AB - Determination of human core temperature has a long history. Since Antiquity, the significance of normal and abnormal body temperatures has been the subject of various interpretations. In this respect, theories based on humoral pathology were replaced by more scientific concepts in the 19th and 20th centuries. Objective measurement and comparison could be performed only after the invention of the thermometer and the introduction of temperature scales. Sanctorius and subsequently Boerhaave and others emphasized the use of measurement of body temperature in the clinic, but its importance was not accepted generally until the late 19th century. Many physicians and scientists have contributed to the progress of thermometry; however, the creation of a firmer scientific basis for clinical thermometry is usually attributed to Wunderlich. PMID- 9340542 TI - [Syncopes during simultaneous use of terfenadine and itraconazole]. AB - A 36-year-old female was given terfenadine 120 mg/day for hay fever, and itraconazole 100 mg twice daily for mycosis. Nine days after starting these drugs, she had several episodes of syncope. The ECG showed a long QT interval and torsades de pointes. The drugs were withdrawn and the patient temporarily received an infusion of isoprenaline, after which the QT interval returned to normal and no further episodes of torsades de pointes occurred. No other causes than the two drugs were found to explain these episodes. PMID- 9340544 TI - [Clinical thermometry. II. Current dilemmas]. AB - The relevance of measuring core temperature for diagnosis and treatment of various diseases is generally acknowledged nowadays. Despite introduction of new techniques and markedly improved understanding of body temperature regulation and the pathophysiology of fever, several dilemmas remain to be elucidated in clinical thermometry. In the measurement and interpretation of body temperature, the many variables that influence core temperature, the site of temperature registration and the significance of an elevated or decreased core temperature should be taken into account. With every type of thermometer, good calibration and an adequate recording technique remain pivotal to obtain reliable and reproducible results. PMID- 9340545 TI - [Comparison of regional formularies for general practitioners]. AB - OBJECTIVE: To inventory the similarities and differences of the contents of regional general practice formularies. DESIGN: Descriptive. SETTING: Department of General Practice Medicine, University of Groningen, the Netherlands. METHOD: Through personal contacts and an appeal in the journal Medisch Contact, 14 formularies were acquired. Of these, seven recent, still valid Dutch general practice formularies were compared as to pharmacotherapy and health problems listed. RESULTS: Volume and contents of these seven formularies showed major, inexplicable differences with regard to pharmacotherapy, while the choice of the health problems listed was not argued anywhere, and not based on prevalence figures. CONCLUSION: The regional general practice formularies examined showed little similarity in the drugs and health problems listed. PMID- 9340546 TI - [A patient with alveolar echinococcosis (Echinococcus multilocularis infection]. AB - In a 45-year-old Swiss male, who had been living in the Netherlands for 20 years, alveolar echinococcosis was diagnosed. He had probably been infected during his youth in Switzerland. His illness became symptomatic more than 20 years later. The diagnosis was reached by microscopic examination of material obtained from a necrotic mass in the liver. Imaging revealed that the disease had spread diffusely throughout the liver, spleen and abdomen. Curative resection was impossible. Percutaneous drainage of the hepatic necrotic mass was complicated by a bacterial infection for which he was treated with antibiotics. Treatment with high doses of albendazole resulted in considerable improvement. The patient represents the first case of Echinococcus multilocularis infection diagnosed in the Netherlands. PMID- 9340547 TI - [Possible effectiveness of mass screening of the population for cervical cancer]. PMID- 9340548 TI - [Possible effectiveness of mass screening of the population for cervical cancer]. PMID- 9340550 TI - [Hot spots: metastases or osteoporosis?]. AB - Radionuclide bone scintigraphy is a sensitive but not a specific technique for the diagnosis of bone metastases. Three patients, a man of 67 on steroid therapy because of chronic obstructive pulmonary disease and prostate cancer, a woman of 76 on steroid therapy because of temporal arteritis and a 50-year-old man with pain in the back, had bone scintigrams showing abnormal uptake (hot spots). There were delays of months before the diagnosis of osteoporosis instead of bone metastases was made. Abnormal uptake on a bone scintigram is often non-specific and requires further evaluation using supplementary diagnostic techniques. PMID- 9340549 TI - [Gluten as a food additive in the Netherlands: cheese prohibited for colic patients?]. PMID- 9340551 TI - [The updated guideline 'Gastric complaints' of the Dutch College of General Practitioners; a reaction from a general practitioner]. AB - Three working diagnoses in gastric complaints are presented: aspecific gastric complaints, ulcer complaints, reflux complaints; these assist the general practitioner in formulating the initial diagnostic and pharmacological interventions and in evaluating the effects of therapy. The guidelines state that eradication treatment of Helicobacter pylori is indicated only in gastritis caused by H. pylori, grade IV bulbitis, a duodenal bulb malformation or peptic ulcers. PMID- 9340552 TI - [The updated guideline 'Gastric complaints' o the Dutch College of General Practitioners; a reaction from a gastroenterologist]. AB - This update has been well thought-out, and is a useful basis for further investigation in general practice. Further simplification may enhance its usefulness, however, e.g. by reducing the proposed three working diagnoses (aspecific gastric complaints, ulcer complaints, reflux complaints) to two: reflux complaints and gastric complaints. In addition, medication in the guideline might be restricted to acid inhibitory treatment and early serological determination of Helicobacter pylori infection might be included. PMID- 9340553 TI - [Diagnosis and treatment of unilocular hydatid disease (Echinococcus granulosus infection)]. AB - Unilocular hydatid disease occurs in humans after oral intake of eggs of the dog tapeworm, Echinococcus granulosus. Cysts develop mostly in the liver, but sometimes also in the lung. The diagnosis of Echinococcus infection is based on history-taking, physical examination, ultrasound and CT examination and serological testing; the diagnosis is confirmed by parasitological examination of cystic fluid. Treatment until some 15 years ago consisted in operation. Subsequently, treatment with initially mebendazole, later with albendazole or with percutaneous drainage (puncture, aspiration, injection of a scolicidal, respiration (PAIR)), or with combinations of the same, became accepted forms of management. The PAIR method is reported to give high proportions of success, low proportions of recurrence and few complications. However, adequate evaluation is not yet possible because of the short follow-up period. For the prevention of leakage it is recommended to perform the PAIR method with a transhepatic puncture under continuous ultrasonographic or CT guidance; for avoidance of recurrences, one week's pretreatment and 1-4 weeks' after-treatment with albendazole are recommended. The results of albendazole monotherapy are hard to predict and highly variable: success: 30-88% (median: 71%); failure: 22-32% (median: 25%); recurrences: 9.5-31% (median: 16%). Both albendazole therapy and the PAIR management should be followed by protracted follow-up to check regression of cysts and detect recurrences. It is not clear which treatment is the best one. PMID- 9340554 TI - [Summary of the updated guideline 'Gastric Complaints' of the Dutch College of General Practitioners]. AB - The Dutch College of General Practitioners recently updated its guideline 'Gastric complaints'. The main change concerns Helicobacter pylori infection diagnosis and treatment. In cases of functional dyspepsia with Helicobacter infection antibiotic eradication treatment is not advised. In patients with gastric complaints, three working diagnoses are presented: aspecific gastric complaints, ulcer complaints, reflux complaints. Endoscopy is given a more prominent role than in the previous version of the guideline. Criteria are given for discontinuation of unmotivated long-term use of antacid medication. PMID- 9340555 TI - [Plastic surgery as a last resort in lichen sclerosus]. AB - Lichen sclerosus, formerly called lichen sclerosus et atrophicus, is a chronic skin disease manifesting itself mostly in the perineal region and often associated with itching. Characteristic elements are well-defined depigmentation and degeneration of the skin sometimes showing haemorrhagic bullae or teleangiectases. The skin grows thinner and shrinks. Malignant degeneration is rare. The prevalence is 1:300 to 1:1000. The condition occurs more often in females than in males and more often in adults than in children. Drug treatment (symptomatic) comprises local application of corticosteroids, anaesthetics and/or sex hormones. In case of insufficient response, cryotherapy is a good alternative. Chemical and surgical neurotomy are also sometimes applied, with fairly poor results. In refractory symptoms, excision of the affected skin, possibly with transplantation using a pedicled skin flap, may lead to mitigation. PMID- 9340556 TI - [Better postoperative pain management in children by introduction of guidelines; a prospective study]. AB - OBJECTIVE: To investigate the influence of recommendations on the quality of postoperative pain management in children. DESIGN: Prospective. SETTING: University Hospital Groningen, the Netherlands. METHOD: After interdisciplinary recommendations on postoperative pain were developed, the quality of postoperative pain management was investigated before implementation (phase I; n = 50 children aged 0-14 who underwent elective surgery), three months after the implementation (phase II; n = 51), and nine months later (phase III; n = 50). Quality was defined by a pain score (for ages 0 to 4 with the 'Children's Hospital of Eastern Ontario pain scale' (CHEOPS) and for ages 4-14 with the Oucher scale) and the prescription of analgetics: kind, dose, frequency, prescription by anaesthetist and doctor on the ward. Pain was scored every 2 hours during the first 24 hours after surgery. A CHEOPS score < or = 6 an Oucher score < or = 50 was defined as adequate; higher scores were defined as inadequate. RESULTS: Pain measurement showed a statistically significant improvement of pain scores in time (phase II and III compared with phase I: odds ratio: 2.5; 95% confidence interval: 1.03-6.00; p < 0.01). Searching for factors that could be responsible for this improvement, like medication, we found no statistically significant differences in everyday practice in phase II and III compared with phase I. However, children who could score their pain by self report (Oucher) showed the best results in all 3 phases of the study. The youngest children, i.e. less than 6 months old, showed inadequate results during the whole study. The greatest improvement in time during the first 12 hours was seen in the group of children older than 6 months. The recommendations were followed more strictly in younger children, and when continuous morphine was given. CONCLUSION: Pain scores in children improved after the introduction of recommendations on postoperative pain. However, the improvement could not be attributed to factors like medication. Factors like a change in attitude towards pain could be responsible for this change. PMID- 9340557 TI - [Blue toes and kidney insufficiency]. AB - Three patients, two women aged 73 and 70 and one man aged 58 years, were known with hypertension and (or) coronary disease. They developed renal insufficiency and purple toes due to cholesterol crystal embolisation. In two of the three patients invasive procedures (femoropopliteal bypass surgery and replacement of the aortic valve, respectively) had provoked the embolisation process. Growing awareness of symptoms such as purple toes is important as the increasing use of intravascular procedures will lead to higher incidence of this syndrome with renal insufficiency as the most severe clinical complication. PMID- 9340558 TI - [European registration of medicines: pros and cons for prescriber and patient]. AB - Since the European Medicines Evaluation Agency (EMEA) in London started its activities, the registration of medicines in the European Union (EU) has gained momentum, and its relevance to national drug registration procedures is steadily increasing. Its main objectives are centralization and standardization of assessments and promoting free trade within the EU. From its early beginning the EMEA has promoted transparency and full disclosure of expert opinions. Since the EMEA is largely operating beyond the scope of the average prescriber and patient, an attempt was made at improving information, based on interviews of a number of responsible or interested persons and parties. Though the overall opinion seems favourable, the quality of the assessment reports has notably improved, and the implementation of decisions has gained speed, this has made little impact on medical practice. The approach towards collating reports of adverse drug reactions has been divisive, and feedback to reporting physicians has been largely neglected. The eventual introduction of the Eudrawatch database and further improvements in transparency will hopefully correct these initial problems. So far as legally possible national drug agencies should follow the example of EMEA by offering public disclosure of documents and expert opinions. PMID- 9340559 TI - [AIDS; new developments. I. HIV tests: time for a more active policy]. AB - Until recently, an HIV test was of limited importance for persons who had been at risk for HIV as effective drugs were not available. With the advent of a new generation of antiretroviral drugs with beneficial effect after early treatment, knowledge of the HIV serostatus has become of more importance to asymptomatic persons. There is no need for a mass screening campaign however, as there is no cure as yet. PMID- 9340560 TI - [AIDS; new developments. II. Treatment of HIV infection]. AB - The currently available anti-HIV drugs can be subdivided according to the mechanisms of action into two main groups, viz, reverse transcriptase (RT) inhibitors and protease inhibitors; the former may be subdivided into nucleoside inhibitors and non-nucleoside inhibitors of reverse transcriptase. Combination therapy is preferable to monotherapy because of resistance problems. The 'ideal' combination consists of two RT inhibitors plus one protease inhibitor. Of the RT inhibitors, zidovudine is to be preferred because it slows the development of AIDS dementia. Other RT inhibitors are didanosine, zalcitabine, stavudine and lamivudine. As regards the protease inhibitors; in view of the development of resistance, it is advised to prescribe ritonavir, indinavir or saquinavir. The antiretroviral management should be adjusted as soon as signs appear of toxicity or failure of the treatment due to resistance or poor compliance. PMID- 9340561 TI - [AIDS; new developments. III. Predictive value of the quantity of HIV-RNA for the course of the HIV infection and the effect of the treatment]. AB - Standardized tests to measure the quantity of HIV-RNA in blood have been available since early 1995. Individual differences in quantity of HIV-RNA in blood are determined mainly by differences in virus production. Most of the HIV production takes place in infected and activated CD4+ T lymphocytes. The concentration of HIV-RNA in plasma or serum is an important predictor of AIDS and death, in the early stage of the infection even the only one. High concentrations (> or = 10(4) HIV-RNA copies/ml) in the early stage of the infection increase the probability of AIDS from 2.5 to 5 times that with concentrations < 10(4) HIV-RNA copies/ml. The degree of reduction of the HIV-RNA concentration during anti-HIV therapy is of great predictive value for the clinical course. The percentage of persons in whom the amount of HIV-RNA decreases to below the limit of detectability is a good second indicator of a favourable prognosis. Consequently, anti-HIV treatment should be started as early as possible in the infection, and not postponed until the first symptoms of immunodeficiency occur; only then can the virus production be kept at a minimal level as early and as long as possible. The current guideline reads: determination of HIV-RNA concentrations in serum or plasma should be part of the standard clinical practice in monitoring HIV infected persons. Treatment is started as soon as occasioned by the quantity of HIV-RNA (limit: 10,000 copies/ml), the CD4+ cell count (limit: 500 x 10(6)/l) or the onset of HIV-related symptoms. The quantity of HIV-RNA should diminish substantially during the first three months of treatment. If this is the case, one checkup every three months will suffice. If the HIV-RNA concentration does not decrease sufficiently, resistance or poor compliance may be involved. PMID- 9340562 TI - [AIDS; new developments. IV. Changes in primary medical care due to new possibilities of treating HIV-infected patients]. AB - A combination treatment has recently become available for HIV-infected patients; it consists of two reverse transcriptase inhibitors and one protease inhibitor. This combination therapy has consequences for primary medical care as regards diagnostics, treatment, follow-up and counselling of patients with HIV and AIDS, education of patients requiring the HIV test, information of seropositive patients not yet being treated and the cooperation and task distribution between GPs and AIDS specialists. The new combination method creates new problems for patients, such as medicalization of their lives, side effects and compliance; it also affects their prospects (social reintegration, resumed occupational activity). In the medical management, the focus shifts to early start of treatment, stimulating compliance, monitoring of effect and side effects, if any, of the treatment and coping with the patients' uncertainty about durability of the effect. If the new treatment proves to be successful in the long run as well, HIV infection/AIDS will become a chronic disease, so that the needs of care and the care methods will increasingly resemble those of other chronic diseases. There will be a growing need of extramural care, leading to new requirements regarding the study of medicine and postgraduate instruction of GPs. PMID- 9340563 TI - [Deep venous thrombosis of the right leg caused by compression by an abdominal aortic aneurysm]. AB - A 67-year-old man was admitted because of suspected deep venous thrombosis of the right leg. At sonography, CT of the abdomen and aortography it became obvious that compression of the right iliac vein by an abdominal aortic aneurysm was responsible for the deep venous thrombosis. This has only been reported before twice. In the case of deep venous thrombosis or oedema of the legs an abdominal aortic aneurysm needs to be excluded. PMID- 9340564 TI - [Is the term 'registration' applicable to homeopathic medicines?]. PMID- 9340565 TI - [What prevalence of diabetes or stored carbohydrate tolerance is found when specifically looking for glycosuria at any random time of the day in addition to determining blood sugar levels?]. PMID- 9340566 TI - [Incidence of cancer near Schipol during 1988-1993]. PMID- 9340567 TI - [Longterm sedation with propofol is contraindicated in children]. PMID- 9340568 TI - [Prevention of diphtheria]. PMID- 9340569 TI - Clinical privileging for APNs. PMID- 9340570 TI - [Morphological aspects of the effect of magnesium orotate therapy on changes in the vascular walls induced by cholesterol-rich diet]. AB - Heart and blood vessel disease are one of the leading causes of death, so their prevention and therapy are very important. In the present study the effects of magnesium chloride, magnesium orotate and orotic acid were tested. New Zealand rabbits were fed with enriched (2%) cholesterol diet during 112 days: starting with day 56 all rabbits were treated with MgCl2, Mg-orotate or orotic acid (orally). Aortas, coronaries, renal and femoral arteries were removed and evaluated by morphological and morphometric methods. Atherosclerotic alterations in each vessel could be influenced moderately by Mg-chloride, quite well by orotic acid and excellently by Mg-orotate. From these results one can conclude that orotic acid and Mg-orotate have a beneficial effect in the prevention and therapy of heart and vessels diseases. PMID- 9340571 TI - [The role of oxidative stress and the preventive effect of free radical scavengers in arteriosclerosis]. AB - The role of oxidative stress in the development of arteriosclerosis is well established. This pathogenetic explanation unificates in itself the lipid and thrombotic theories. The authors summarize the most substantial literary data in this relation, they discuss in details those therapic methods, in which the natural and synthetic antioxidants are involved as preventive drugs in the development and consequences of arteriosclerosis. Thus the effects of the dihydroquinoline type antioxidants as well as those of Vitamins A, C and E are discussed partly in experimental, partly in clinical studies. The authors conclude on the basis of own and literary data that the application of antioxidants could decrease the blood vessel alterations produced by arteriosclerosis, as well as the pathological tissue alterations developed in the consequences of ischaemia. PMID- 9340572 TI - [Biologic effect of LDL binding and intracellular degradation in monocytes from patients with hypercholesterolemia]. AB - The granulocytes from elderly patients were investigated, in previous studies, with FMLP and it was found that the postreceptor signal, the inositol phosphate production and inositol phosphate dependent calcium signal were markedly reduced. It was observed that the 125I LDL binding was slightly reduced while the intracellular degradation of the LDL and endogenous cholesterol synthesis inhibitory effect was significantly decreased on monocytes of patients with non insulin dependent diabetes mellitus. It was suggested that of in patients suffering from NIDDM with hypercholesterolemia the LDL receptor numbers of monocytes are close to normal, while the post receptor signal transmission is damaged. In this study the monocytes from 12 patients with hypercholesterolemia were investigated before and after LDL treatment and were compared to the 11 age matched healthy volunteer control patients. The cells were stimulated with LDL and chemotactic peptide FMLP. The postreceptor signal mechanism in monocytes was investigated. According to the results the inositol phosphate level of the patient group decreased independently from the stimulus. The LDL induced IP3 and Ca2+ level elevation was PT resistant both in the control and in the patients group. PMID- 9340573 TI - [Mononuclear phagocytes in peripheral blood as cellular markers of lipid and lipoprotein metabolism]. AB - The functional specialization related to tissue-specific immune surveillance mononuclear phagocyte differentiation is characterized by and increased by and expression of scavenger receptors and synthesis of apo E. Apo E heterogeneity related to different patterns of the cellular uptake and metabolism of lipoproteins represents an interesting genetical model for the analysis of the interaction between cellular lipid metabolism and monocyte differentiation. In a preliminary study of the expression of differentiation dependent antigens on monocytes of hypercholesterolemic patients they observed an altered expression density of maturation associated antigens on monocytes from patients with familial hypercholesterolaemia. The abnormalities of mononuclear phagocytes may correlate with the cardiovascular state within hypercholesterolemic patients. This stimulated the authors to perform a more detailed study of the immunological phenotypes of circulating blood monocytes and their subpopulations as potential diagnostic parameters which may correlate to the atherogenic risk, status of atherosclerosis or therapy response in patients with hypercholesterolemia. PMID- 9340574 TI - [Characteristics of apolipoprotein polymorphism in a Hungarian population]. AB - One of the main risk factors of ischaemic heart disease is the dyslipidaemia. In the development of altered lipid metabolism genetic factors play a significant role. The hereditary backgrounds have to be known in order to reveal the most important possibilities in diagnostics and prevention. PMID- 9340576 TI - [Granulocyte function and lipid peroxidation in untreated patients with hyperlipoproteinemia]. AB - The fatty acid composition in free fatty acid and phospholipid fraction of plasma in untreated mildly hyperlipidemic patients were determined. The general trend was an increase in saturation in both free and phospholipid fractions of plasma in patients compared with that of healthy controls. Furthermore, arachidonic acid, precursor of formation of prostaglandins and leukotrienes was detected in significantly lower amount in plasma of mildly and untreated hyperlipidemic patients. These fatty acid abnormalities were connected with the increased lipidperoxidation in plasma lipids and in both resting and stimulated granulocytes. PMID- 9340577 TI - [Lipoprotein (a) studies in thyroid diseases]. AB - High lipoprotein (a) levels have been established as a risk factor for atherosclerosis. The physiological and pathological function of lipoprotein (a) and the effect of other factors of lipoprotein (a) plasma concentrations are not well known. The role of thyroid hormones concerning Lp(a) plasma concentrations are not fully cleared. The Lp(a) levels are genetically determined, relatively stable, while the thyroid hormones react differently. 30 hyperthyroid and 29 hypothyroid patients were studied. The thyroid status and lipid parameters were investigated. A significant correlation was observed between the cholesterol, triglyceride, LDL-cholesterol, Apolipoprotein B and the thyroid hormone levels. The HDL-cholesterol and Apolipoprotein A correlated inversely with the thyroid hormone levels. Data obtained on the Lp(a) and thyroid hormone was in contrast with those of other studies. Hyperthyroid patients were increased Lp(a) concentrations as compared those of euthyroid subjects. In severe thyreotoxicosis (especially with high levels of antithyroid antibodies) elevated Lp(a) plasma concentrations were found. In hypothyroid patients the high levels of Lp(a) were measured as described in previous studies. Authors concluded that further studies are needed to investigate this phenomenon. PMID- 9340575 TI - [Abnormal function of lipoprotein receptors in the monocytes of hypercholesteremic patients]. AB - The familial hypercholesterinemia (HCh) is as a genetically determined disorder. The genetical damage and functional abnormalities of the LDL receptors lead to familial Hch. The LDL plays an important role in cholesterol metabolism. They carry cholesterol which metabolizes through specific and scavenger LDL receptors. The ApoB100 particle of LDL binds to the receptors, internalizated, and digested, and the remaining free cholesterol regulates the intracellular cholesterol synthesis. It inhibits the key enzyme, HMG-CoA reductase and decreases the LDL receptor synthesis and increases cholesterol esterification. These mechanism can prevent the cholesterol accumulation of the cells. The aim of the present study was to clarify the activity and number of the LDL receptor, to study the LDL binding and degradation and to evaluate how the intracellular cholesterol can regulate the synthesis in patients with HCh. 58 pts with HCh and their monocytes were investigated, because the monocyte derived macrophages contained both specific and scavenger receptors. Monocytes of the pts were compared to the healthy individual controls. From the results it could be recognized--that the decreased binding to the specific LDL receptors only at 6 pts cholesterol synthesis was elevated in HCh pts group, while the synthesis inhibition induced by 50 micrograms LDL was decreased. The presented experimental results suggested that the decreased binding ability to LDL receptors is a rare cause of cholesterol abnormalities, while during the intracellular degradation process more metabolic steps can be damaged in patients with HCh. PMID- 9340578 TI - [In vitro study of endothelium-dependent relaxation and contraction of human atherosclerotic femoral artery]. AB - According to present knowledge, altered arterial reactivity associated with hypertension, atherosclerosis and hypercholesterolemia is related to impaired release of endothelial derived relaxing factor (EDRF). Impaired relaxation followed by enhanced vasoconstriction leads to a well known clinical entities such as unstable angina, acut myocardial infarction. Impairment of EDRF may also account for smooth muscle cell proliferation and migration. Aim of present study was to examine the endothelial dependent response during in vitro conditions in human femoral arteries taken from bypass operation and leg amputation. Examining the contractility, the effect was modulated with nifedipin, EDTA and pertussis toxin, respectively. Endothelium dependent relaxation to acethylcholin and histamine were markedly diminished, while those to calcium ionophore were maintained throughout the study. These results suggest that at least two or more receptor-coupled system may be involved in generation of EDRF. However, direct relaxation of femoral artery to nitrovasodilatators (nitroglycerine) were comparable between control and atherosclerotic artery. Another striking change in atherosclerotic artery was the increased sensitivity to the vasoconstrictions. To eluciadate to exact biochemical mechanism underlying the endothelial dysfunction may help to develop a new vasodilatator drug. PMID- 9340579 TI - [Intracellular signal transmission of low density lipoprotein receptors in human monocytes]. AB - The authors found that in human monocytes administered low density lipoprotein in doses of 50 micrograms had optimal inhibition of endogenous cholesterol synthesis which measured by [14C] acetate incorporation. There was not effect of pertussis toxin and phorbol myristate acetate on the inhibiton of endogenous cholesterol synthesis, whereas calcium channel blocker verapamil and phospholipase A2 inhibitor chloroquine decreased it. In contrast, the protein kinase C-stimulant phorbol myristate acetate alone had effects as LDL, but the protein kinase C inhibitor H-7 had antagonist effect against LDL. Inositol phosphate generation was induced by administration of LDL in doses of 50 micrograms, which was pertussis toxin insensitive. The calcium signal was not also pertussis toxin sensitive, while occurred an intensive protein kinase C activation by administration of LDL. In signal transduction of monocytes activated by LDL may be an important role of the opening of calcium channels and activation of two enzymes, such as phospholipase A2 and protein kinase C. PMID- 9340580 TI - [2-year results of leg amputation in Hungary based on a nationwide data base]. AB - Authors review nation-wide hospital data of amputees over two years in order to make comparison with similar data gathered about 20 years ago. Data were provided by the National Medical Records Centre and processed by own developed programmes. The quality level of data validity is slightly criticised. The cause of amputation was most often vascular disease, amputees were elderly and large majority of amputation surgery was carried out on the lower limb. The rate of below-knee amputation has gone up favourably over the 20 years, but there are large regional differences within the country. Mortality parameters remarkably exceed those of foreign countries. Regarding compromised data accuracy, still there are ways of exploiting data in favour of quality improvement of care, e.g. improve below-knee amputation rate, reduce mortality. Publication of data can be of bench-marking importance for hospitals by enabling them to compare own results with those of other hospitals, as well as develop and improve performance. PMID- 9340581 TI - [Management of hemodynamically significant fetal arrhythmias]. AB - Between January 1, 1993, and April 30, 1996, authors treated 23 fetuses with severe rhythm disturbances in their Department. The correct diagnosis was made by fetal echocardiography. They had 15 tachyarrhythmic and 8 bradyarrhythmic patients. They found hydrops fetus at 7 patients because of atrial flutter (2 fetuses), supraventricular tachycardia (4 fetuses) and severe bradycardia (1 fetus). They treated successfully 13 patients with antiarrhythmic therapy given to the mother. They had 1 intrauterine death (treated because of bradycardia) and 1 neonatal death (hydropic because of supraventricular tachycardia). The causes of severe bradycardia were maternal antibody (3 fetuses), cardiac malformation (3 fetuses) and large number of blocked atrial extrasystoles. The prognosis of fetal tachycardia is good even in cases of fetal hydrops. The prognosis of bradycardia due to heart abnormalities is poor. PMID- 9340582 TI - [Customary antidepressive treatment in Hungary: moclobemid therapy in everyday practice]. AB - Treatment data with moclobemide have been obtained from clinical practices of 91 Hungarian psychiatrists. Three hundred and twenty seven patients were included in the multicenter postmarketing surveillance study and observed during an 8 week treatment period. The mean age of the patients was 50 years, 71% were woman and 33% were outpatients. Sixty-five percent had preexisting depression. Previous antidepressant treatment for this episode had been received by 45%, with the most common reasons for change to moclobemide being inadequate response. Moclobemide was indicated in patients with mainly moderate to severe major depression and dysthymia. The frequency of comorbidity was 44%. The dosage of moclobemide was usually started at 300 or 450 mg/day and increased to 600 mg/day only in some patients. Three divided daily dosages were administered in 53% of the patients. Nine percent discontinued the treatment before the end of the observation period. 87% had no reported adverse events. The most common adverse events were sedation (3.7%), agitation (3%) and insomnia (3%). Concommitant medication from all major drug groups were taken by 250 (76.6%) patients, with no adverse interactions. The current therapeutic habits of the Hungarian psychiatrists in treatment of depression were also revealed. The findings in the normal clinical psychiatric practice suggest that moclobemide is a safe drug with excellent tolerability in the treatment of various subgroups of depressive disorders and especially useful in patients with coexisting physical illness. Moclobemide is a favourable alternative to the modern effective antidepressant agents in the Hungarian clinical practice. PMID- 9340583 TI - [The use of Wallstent in malignant constrictions of the gastrointestinal tract]. AB - Prosthesis implantation in malignant oesophageal stenosis, postoperative gastric outlet obstruction and jejunal stenosis is a quick and efficient method. The expansile metal stents are a new alternative to conventional plastic prosthesis. The Wallstent is made of surgical steel alloy elements braided in tubular fashion. The Wallstent is delivered in a small diameter device, but it expands to a much larger size after placement than the inner diameter of plastic stent. The small predilatations diameter makes implantation of the Wallstent less prone to cause complications or severe discomfort to the patient. The authors implanted seven Wallstents in six patients. One patient had oesophageal cancer, one ischaemic jejunal stenosis and four jejunal stenosis due to extraluminal tumour recurrences following total gastrectomy for cancer. The authors discuss in detail the usefulness of the Wallstent implantation in two cases with malignant disease of the oesophagus and jejunum. They remained symptoms free for 7 and 10 months after Wallstent implantation until death. Despite their higher initial cost, the metal stents are cost effective because of the absence of early and severe complications and the decrease in the hospitalization. PMID- 9340584 TI - [Androgen-producing adrenocortical adenoma in childhood. Pitfalls of differential diagnosis]. AB - A two-year-old girl presented with clitoromegaly and an abdominal mass. Diagnostic procedures including sonography, computerized tomography, scintigraphy and measurement of catecholamines in urine excluded neuroblastoma, but suspected Wilms-tumor. Before completing the steroid measurements therapy was initiated according to Wilms-tumor (preoperative cytostatic therapy followed by surgical removal of the tumor). Morphology of the tumor, the serum and urinary steroid profile proved a benign adrenocortical adenoma producing mainly delta 5-steroids including the weak androgen, dehydroepiandrosterone. PMID- 9340585 TI - [Could Helicobacter pylori cause reactive arthro-osteitis?]. PMID- 9340586 TI - [Anatomy of the brachial plexus]. AB - The early development of the brachial plexus shows that it is formed of a dorsal branch supplying the extensor muscles and a ventral branch for the flexor muscles. Although the network becomes more and more complex, a basic pattern is generally preserved. This basic pattern, as well as variants and the topography of the lateral neck triangle, is described. PMID- 9340587 TI - [Brachial plexus injuries in adults]. PMID- 9340588 TI - [Diagnosis and surgical indications of traumatic brachial plexus lesions from the neurosurgery viewpoint]. AB - The precise preoperative clinical and electrophysiological evaluation of the brachial plexus as well as an exact radiological evaluation are the keystones for the treatment of traumatic injuries of the brachial plexus. Furthermore, surgical management and prognosis of traction injuries of the brachial plexus depend on the accurate diagnosis of root avulsion from the spinal cord. Myelography, myelo computed tomography and recently magnetic resonance imaging are the main radiological methods for preoperative diagnose of cervical root avulsions. Surgical experience shows that in may cases, extraspinal findings diverge from intradural findings. Consequently, only correlation with the intradural surgical findings will allow us to define the factual accuracy of myelo-CT and MRI studies. Accuracy of the preoperative myelo-CT based diagnosis related to the intraoperative intradural findings was 85% On the other hand, MRI showed an accuracy of only 52%. Therefore, myelo-CT scans with 1 to 3 mm axial slices proves to be the most reliable method to evaluate preoperatively the presence of complete or partial root avulsion in traumatic brachial plexus injuries. However in 15% of the cases preoperative exact radiological diagnosis is unfortunately not reliable. In these special cases intraspinal surgical exposure of the cervical roots will provide the accurate diagnosis of root avulsion. Accurate clinical evaluation and exact assessment of intraspinal root avulsion simplify enormously the decision concerning the choice of donor nerves for transplantation and/or neurotization during brachial plexus surgery. PMID- 9340589 TI - [Nerve transfer (neurotization) for functional reconstruction of arm functions in cervical root avulsions]. AB - Neurotization procedures represent an important therapeutic option in patients with complete root avulsion due to traumatic brachial plexus injuries. A variety of normal donor nerves can be used, including intercostal nerves, accessory nerve, parts of the cervical plexus, phrenic nerve, and/or the contralateral C7 nerve root. The reconstructive neurotization procedure should be performed within 3-6 months following the trauma and, when successful, can lead to substantial improvement in motor function for the shoulder and upper arm and in sensory function for the forearm, hand and finger region. Neurotization can also be combined with neurolysis and/or transplantation to restore upper limb motor and sensory function in order to achieve greater therapeutic benefit for patients with posttraumatic brachial plexus lesions. PMID- 9340590 TI - [Nerve transplantation and neurolysis of the brachial plexus after post-traumatic lesions]. AB - Autologous transplantation in brachial plexus injuries has become significantly more efficacious through increased knowledge of the microtopographic anatomy of the fascicle structures and the refinement of microneurosurgical techniques. Recovery of shoulder and upper arm functions can be achieved with autologous transplantation in the majority of patients under optimized conditions. However, attempts to restore forearm, hand and finger functions in this way have been disappointing so far. Therapeutic success is primarily governed by early decision making, selection of adequate surgical strategies and intensive and longlasting postoperative management. PMID- 9340591 TI - [Pain management after post-traumatic brachial plexus lesions. Conservative and surgical therapy possibilities]. AB - The occurrence of sever pain is one of the most disabling symptoms after the traumatic lesion of the brachial plexus. Avulsion of one or more cervical roots of the brachial plexus is the main cause of severe pain, known as deafferentation pain. Lesion of the dorsal horn of the cervical spinal cord due to root avulsion may lead to important pathological changes and scarring that are responsible for the induction of pain sensations. Different medical and surgical treatment modalities have been established to relief such pain after brachial plexus injury. In contrast to drug therapy, which usually offers only limited benefit, surgical treatment over the last years has shown positive results. Coagulation of the dorsal root entry zone (DREZ) is one of the most efficient surgical treatments for these patients. Understanding of the pathophysiological changes and different pain mechanisms induced by traumatic injury of the brachial plexus is fundamental for the planning and step-wise treatment of such patients. PMID- 9340592 TI - [Historical development of muscle replacement operations in brachial paralysis]. PMID- 9340593 TI - [Shoulder arthrodesis in deltoid paralysis]. AB - After loss of active glenohumeral motion the arm cannot be positioned in space and function is impaired. Fusion of the glenohumeral joint creates a new platform for the movement of elbow, hand and fingers and leads to an improvement of function. The use of the reconstructive plate makes the operation easier. Shoulder fusion is especially suited if the function of elbow and hand is well preserved and if the patient is physically active. But fusion is definite and the options of muscle transfer operations and above elbow amputation have to be considered. PMID- 9340594 TI - [Trapezius transfer in deltoid paralysis]. AB - In most cases the genesis of brachial plexus palsy is traumatic, often because of bike accidents. If physiotherapy and neurosurgical procedures such as nerve repair do not have the desired outcome, muscle transfer operations are possible. The results of our favored transfer of the trapezius muscle to compensate paralysis of the deltoid muscle will be presented. Preoperatively radiological, clinical and electromyographic examinations are necessary. Our results are based upon the clinical and radiological check ups and the subjective assessment of the patients. Thirty-one patients (7 female, 24 male) underwent a trapezius transfer between March 1994 and December 1996. The average age was 29 years (range 18-46 years). We performed the operations using a modification of Saha's technique. With the patient in lateral decubitus position and protection of the opposite plexus, a sagital skin incision is the first step, followed by the preparation of trapezius and deltoid muscle as well as the bony parts of the shoulder (acromion, clavicle, scapular spine). The deltoid origin is cut from the lateral third of the clavicle, the acromion and the lateral half of the scapular spine. The next step is transection of the roof of the acromion and the lateral clavicle. After elevation of the remaining trapezius insertions from the clavicle and scapular spine, the proximal humerus is exposured by splitting the partly detached deltoid muscle longitudinally. Then the acromion fragment and humerus are prepared for the bone-to-bone contact. In 90 degrees of abduction the acromion fragment with its trapezius insertion is transferred and fixed to the humerus with two 4.5-mm screws. Finally the deltoid is sutured on the top of the trapezius and the skin is closed over two suction drains. Postoperatively we immobilize the operated arm in an abduction support for 6 weeks. The physiotherapy program starts on the first postoperative day with active training of elbow, hand and fingers and electrostimulation of the transferred trapezius muscle. Six weeks after the procedure we take an X-ray and start with progressive adduction of the arm. The preoperative subluxation of the humeral head was abolished in all cases. We achieved an average increase of active abduction from 7.3 degrees (range 0-45 degrees) preoperatively to 39.2 degrees (range 25 degrees-80 degrees) 1 year after the operation; the increase of forward flexion was from 20 degrees (range 0 degrees-85 degrees) to 43 degrees (range 20 degrees-90 degrees). All patients were satisfied with the improvement of stability and function of the operated shoulder. Finally we can conclude that the trapezius transfer for flail shoulder gives a satisfactory outcome regarding shoulder function and stability as well as the subjective situation of the patients. PMID- 9340595 TI - [Secondary replacement operations for reconstruction of elbow joint function after lesion of the brachial plexus]. AB - Elbow flexion plays a key role in the overall function of the upper extremity. In the case of unilateral complete brachial plexus lesion, restoration of elbow flexion will dramatically increase the patient's chances of regaining bimanual prehension. Furthermore, depending on the type of reconstruction, stability of the glenohumeral joint as well as some supination function of the forearm can be restored to a varying degree at the same time. Depending on the level of brachial plexus lesion and/or reinnervation, different reconstructive procedures are available. In order to select the best treatment option for the patient it is necessary to known the extent of the lesion of the brachial plexus and/or ventral upper arm muscles, to time the operation appropriately, to be aware of all treatment possibilities and to recall the special problems of tendon transfer for brachial plexus patients. Our concept is based on our experience with more than 1100 patients presenting a brachial plexus lesion between 1981 and 1996 and treated in our institution. There were 528 operative revisions of the brachial plexus. Some 225 patients underwent secondary muscle/tendon transfers. In 35 patients elbow flexion was reconstructed by bipolar latissimus dorsi transfer (n = 10), triceps-to-biceps transfer (n = 15), modified flexor/pronator muscle mass proximalization (n = 6) and the multiple-stage free functional muscle transfer after intercostal nerve transfer (n = 4). PMID- 9340596 TI - [What's new in the USA?]. PMID- 9340597 TI - [Hallux valgus]. PMID- 9340599 TI - [Battista Grassi, a zoologist for malaria]. PMID- 9340598 TI - Echinococcus granulosus antigen 5 may be a serine proteinase. PMID- 9340600 TI - [Battista Grassi and malaria]. PMID- 9340601 TI - [The exhibition: an itinerary of the times and the research]. PMID- 9340602 TI - Introduction: the rational for early intervention in cystic fibrosis. PMID- 9340603 TI - [Study of X-ray filter and peak kilovoltage in Fuji Computed Radiography in regard to the detection of simulated pulmonary nodules]. AB - To determine a suitable combination of X-ray filter and tube kilovoltage for Fuji Computed Radiography (FCR), to provide better detection of pulmonary nodules and reduce patient exposure, we compared observer performance with different X-ray filters and tube voltages. Radiographs were obtained with a copper filter backed by aluminium, with a tungsten filter backed by yttrium and aluminium, and with a lead filter backed by yttrium and aluminium, at both 100 kVp and 135 kVp. Observer performance in detecting simulated lung nodules, which were placed on the posterior aspect of a chest phantom, was compared using receiver operating characteristic (ROC) techniques for each combination of the X-ray filter and tube voltage. The results of the study indicated that 1) nodule detection was superior for the images obtained with 135 kVp as compared with 100 kVp; 2) approximately equal detection rates were obtained for the three X-ray filter sets, although results with the tungsten filter were slightly inferior to the other two filters; and 3) the absorbed dose may be reduced by 30% with the use of a lead filter backed by yttrium and aluminium compared with a copper filter backed by aluminium. We conclude that chest radiography with FCR should preferably be conducted with a higher kilovoltage, e.g., 135 kVp rather than 100 kVp, to ensure a higher detection rate of pulmonary nodules and in conjunction with a lead filter backed by yttrium and aluminium to reduce X-ray exposure to the patient. PMID- 9340605 TI - [The success of maternal feeding with very low birth weight premature infants, singletons and twins: a 10-year experience]. AB - There are very few reports about the feasibility of maternal milk feeding in very low birthweight preterm infants (VLBW), especially in twins. Therefore we conducted a cohort retrospective study to evaluate the feeding patterns of the 226 VLBW discharged from our neonatal intensive care unit from 1987 to 1996. Their gestational age was 30 +/- 2.6 weeks, birthweight 1166 +/- 224 g and they were hospitalized for 67 +/- 37 days (means +/- 1 SD). Of the 226 VLBW 49% were males, 39% had birthweight below 10 degrees centile for gestational age and 56% were born to non-residents in our area. There were 181 single births and 45 (20%) multiple births, of which 16 from pregnancies with 3 or more fetuses. Of the total cases 22% were discharged feeding maternal milk (MM) exclusively and 21% on mixed-feeding, maternal + formula milk (FM). Percentages were respectively 23% and 18% for single newborns, 11% and 29% for twins. Singletons and twins were discharged on FM with comparable percentages (59 and 60%). With passing years we have noticed a significant increase (chi square for linear trend < 0.01) for maternal milk feeding. In the last 2 years 49% of singletons and 38% of twins were discharged on MM, 14% and 24% on MM + FM, and only 37% and 38% on FM only. Between singletons and twins there were no statistically significant differences as far as feeding at discharge is concerned. CONCLUSIONS: most mothers, if correctly informed and encouraged, are able to breast-feed, exclusively or partially, their VLBW offspring, including twins, in the first months of life. PMID- 9340604 TI - [Atenolol: its cardiorespiratory effects at birth in a premature infant]. PMID- 9340606 TI - [Syncopal pathology in childhood (I)]. AB - Syncope is a common phenomenon, well-known to all pediatricians: it is defined as a sudden transient loss of consciousness associated with inability to maintain postural tone that is incompatible with a seizure disorder, vertigo, dizziness, coma, shock or other states of altered consciousness. The purpose of this study are to analyse the multiple causes of syncope, to determine the characteristics of pediatric patients with syncope, to define the pathophysiologic mechanisms that result in neurally mediated syncope. PMID- 9340607 TI - [Atherosclerosis in childhood: the role of obesity]. AB - A variety of studies indicates that the process of atherosclerosis begins in childhood and progresses during adulthood. Chronic obesity, inadequate caloric intake, and hypertension and smoke, are associated with an increased cardiovascular disease. The aim of this study is to investigate if the presence of some risk factors during adolescence may involve in accelerated atherosclerosis disease. 50 subjects, median age 11 +/- 0.6 SD (27 females, and 23 males) are admitted to the study. After overnight fasting we have investigated: lipoproteina A (nephelometric test), glycemia and insulin baseline and after load 120', tryglycerides, cholesterolo, apolipoproteina A, B, plasma concentrations. In addition to general medical evaluation, anthropometric measurements of weight, height, blood pressure, BMI, overnight ratio were calculated according to Tanner's charts. The means anthropometric and metabolic values in different groups were compared. One group affected with abdominal obesity state (waist-hips ratio > 0.9), the second with mid obesity condition (waist-hip ratio < 0.9). Tryglycerides, cholesterolo, insulin plasma concentrations in both groups were considered similar. However in the first group higher levels of apolipoproteina A (means 102 + 10.2 SD) and lipoproteina A were demonstrated (P = 0.03 in males, P = 0.01 Statview for Mann Whitney test). Childhood is an important period for the development of the atherosclerosis such as the presence of obesity during this time has a very high likelihood of persisting into adulthood. The severity of obesity in adults is greater in those who were obese as adolescents. In accord with other authors we have not observed abnormal tryglicerides and cholesterolo plasma concentrations, which probably are found in adulthood obesity. We believe indeed the risk factors are different in obesity of childhood, atherosclerosis may be induced by high endogenous insulin secretion and abnormal uptake of lipoprotein. However the potential consequence of excessive insulin secretion could be due in part to insulin effects on recruitment of histiocytosis cells during the development of atheroma and through the modulation of hepatic production and peripheral uptake of lipoproteins. PMID- 9340609 TI - [Adolescent emergencies: the need for active prevention to the spread of STDs (sexually transmitted diseases)]. AB - The authors suggest a program of diffusion of posters which invite young adolescents to pay attention to the symptoms of the S.T.D. They suggest also to found a telephone line where the adolescent can call for any problem or question. PMID- 9340608 TI - [Double phototherapy with Wallaby optic fibers versus conventional phototherapy. Case reports]. AB - The authors have valued the efficacy of the double phototherapy with fiberoptic Wallaby vs conventional phototherapy in 2 groups of term infants, without any complication at birth, utilized respectively as study group and control group. While conventional phototherapy produced a bilirubin reduction of 0.60 +/- 0.26% per hour (with a total reduction of 28.1 +/- 11.1%), the double phototherapy was statistically more effective (p < 0.05) then conventional phototherapy causing a bilirubin reduction of 0.73 +/- 0.28% (with a total reduction of 33.3 +/- 9.5%). At 24 hour after the interruption of the treatment 9 newborns of the study group (36%) and 7 of the control group (28%) presented a rebound effect (increase of the bilirubinemia more than 17 mumol/l), but without a statistical difference (p > 0.05). Our study shows that double phototherapy with Wallaby fiberoptic and conventional phototherapy represent a valid strategy in the treatment of the non haemolytic neonatal hyperbilirubinemia, because, compared to conventional phototherapy, double phototherapy is more effective and reduces the period of the treatment, showing a simple management of the jaundiced newborn. PMID- 9340610 TI - [Portable bladder ultrasonography: a new technic for the urological management of pediatric spinal cord damage]. AB - Noninvasive measurement of bladder volume demonstrates how a new technology can improve the management of pediatric patients with spinal cord injured. The bladder-scan was used to perform volume and post-void residual urine measurement. The aim of our study is to verify the reliability of Bladder Manager Tc 5000 in these patients. PMID- 9340611 TI - [The risk of infection after a tick bite in an endemic area. The experience of the use of an effective protocol on a cohort of Tuscan children]. AB - The authors suggest an executive protocol for the infections due to the sting of a tick in a child. They report the experience they had observing 92 children in the period between spring and autumn in the year 1996. PMID- 9340613 TI - [Subdural effusion during bacterial meningitis]. AB - Twelve children of age ranged from 4 to 34 months with Haemophilus influenzae type b meningitis treated at Meyer Hospital of Florence, were retrospectively reviewed. Eight patients had subdural effusion demonstrated with TC, RM and transfontanellar ultrasonography. All patients are cured without sequelae. PMID- 9340612 TI - [Primary acute acalculous cholecystitis in childhood: a report of 2 clinical cases]. AB - Acute acalculous cholecystitis (AAC) is not frequently encountered in adults and children whether in association with other conditions or above all in primitive form. AAC in infancy, although rare, is well recognized, but its possible presentation is not always kept in mind in considering the differential diagnosis of the acute abdomen. On the other hand, AAC has significant clinical signs and abdominal u.s. scanning usually provides evidence of diagnosis. Only early diagnosis has been shown to limit high morbidity and mortality rates for AAC, since cholecystectomy is the simple procedure of choice for treatment of AAC. We report two cases of primitive AAC one in a six and one in two half years old girls. So we consider the most important features about etiologic factors, pathogenesis, clinical signs and therapy through review of the literature and our personal experience. PMID- 9340614 TI - [The FF complex: a rare congenital malformation]. PMID- 9340616 TI - [The persistent mullerian duct syndrome with transverse testicular ectopia. A hypothesis on the role of mullerian inhibiting factor in the process of testicular migration]. AB - The authors report a rare case of persistent mullerian duct syndrome (PMDS) with transverse testicular ectopia and inguinal hernia in a 2-year-old child with family history for this syndrome. At operation the observation of a very long and thin gubernaculum and extreme mobility of both testes and uterus, which are located in the same hernial sac, allowed the Authors to propose a hypothesis to explain the role of MIF (Mullerian Inhibiting Factor) in testicular descent. Patients with PMDS present a normal outgrowth and migration phases of the gubernaculum but lack of the gubernacular regression phase. These data suggest an important function of the MIF in this phase of testicular descent. PMID- 9340615 TI - [Pseudohyperaldosteronism secondary to licorice poisoning associated with hemorrhagic gastritis]. AB - The case is described of a 6 1/2-year-old child with pseudohyperaldosteronism due to excessive and prolonged liquorice ingestion. The authors debate its differential diagnosis, its physiopathological mechanism (glycyrrhetinic acid, the active metabolite of liquorice, inhibits the conversion of cortisol in cortisone) and its unusual association with haemorrhagic gastritis never observed in the course of liquorice intoxication. PMID- 9340617 TI - [Sandifer's syndrome: a rare form of torticollis in childhood. A report of a patient]. AB - Sandifer syndrome is an uncommon clinical entity characterized by gastroesophageal reflux, torticollis and paroxysmal dystonic postures. For the wide variability in clinical expression it is diagnosed as neurological disease. We report on a 3-year-old patient who presented sudden extensions of the head and neck with tilting of the head one side and severe arching of the spine. It is presented a review of the related literature. PMID- 9340618 TI - [On the problem of modern gestosis terminology]. PMID- 9340619 TI - The value of preoperative knee aspiration: don't ask, don't tell. PMID- 9340620 TI - [Proceedings of the 10th annual session of the working group of the German Society for Pneumology. Nocturnal Respiratory and Circulatory Disorders. Freiburg, 16-18 March 1995]. PMID- 9340621 TI - [Differential diagnosis of seizures in sleep]. AB - Epilepsies and the NREM and REM parasomnias represent the most important sleep related attacks that can occur in conjunction with sleep apnoea. Most sleep related attacks arise from certain sleep stages only, which facilitates their identification. Many attacks can only be classified by thorough observation and description, since they lack special electrophysiological correlates. EEG recordings with at least 16 channels and EMGs from several muscles are necessary for differential diagnosis. Video-documentation often reveals relevant additional information confirming the diagnosis. PMID- 9340622 TI - [Multiple sleep latency test in patients with obstructive snoring]. AB - The objective of our study was to examine the effect of the n-CPAP on day tiredness of patients suffering from obstructive snoring. This effect was objectified by means of the Multiple Sleep Latency Test (MSLT). The MSLT was performed with optimal pressure at 8.00, 10.00, 12.00 and 14.00 hrs. subsequent to the control night and the third CPAP night. The latencies of falling asleep and the sleep stages were determined in accordance with the criteria of Rechtschaffen and Kales. The average latency of falling asleep before therapy was: at 8.00 hrs 9.0 +/- 14.2 min, at 14.00 hrs. 7.2 +/- 6.3 min. The following latencies of falling asleep were observed after the third CPAP night: 8.00 hrs. 14.2 +/- 6.3, 10.00 hrs. 13.4 +/- 6.4, 12.00 hrs. 13.7 +/- 6.4 hrs. 13.7 +/- 6.0 min. This means that after the therapy there was a marked tendency to longer latencies at all 4 points of measurement with significant differences at 12.00 and 14.00 hrs. A comparison of the quality of sleep before and after the therapy yielded an increase in deep sleep and a significant increase in REM density during dream sleep. MSLT enabled objectivation of improved sleep quality and of subjective decrease in day tiredness after CPAP therapy in patients with obstructive snoring. The latency in falling asleep increased at all the points of measurement. Nevertheless, interindividual differences are great, compared with the uniform subjective success of CPAP therapy achieved with these patients. PMID- 9340624 TI - [Restless legs syndrome and periodic leg movements during sleep in patients with sleep apnea--a therapeutic problem?]. AB - Restless-legs syndrome and periodic movements during sleep are associated with sleep apnoea syndrome. Similar to sleep apnoea syndrome, restless-legs syndrome and periodic movements during sleep may cause severe hyposomnia and hypersomnia. Exact diagnosis may partly fail in severe obstructive sleep apnoea syndrome if only cardiorespiratory polygraphy is performed. Simultaneous videorecordings and EMG of mm. tibialis ant. ensure diagnosis. Therapeutic regime may be difficult due to failure or side effects, however. We report on our experience in an one year follow-up of 12 patients with sleep apnoea syndrome and restless-legs syndrome and/or periodic movements during sleep. Despite adequate interdisciplinary initiation of therapy and monitoring, these patients are often subject to therapy changes, failures or side effects. There was no correlation between jerks and the complaints of the patients. PMID- 9340623 TI - [Event related potentials and neuropsychological studies in sleep apnea patients]. AB - Patients with obstructive sleep apnoea syndrome (OSAS) commonly complain about daytime sleepiness and a decline of cognitive functions. Several diagnostic tools have been established to judge objectively vigilance and cognitive impairment. Forty OSAS patients aged between 34 and 74 years were examined via several neuropsychological tests (e.g. vigilance test of the Wiener Testsystem, number connection test ("Zahlenverbindungstest-ZVT") to assess working velocity and information processing, d2-test to rate concentration on exertion) before starting continuous positive airway pressure (CPAP) therapy. In addition, visual evoked event-related potentials (ERPs) were recorded; the P3-component was evaluated. All patients subjectively stated daytime sleepiness and cognitive dysfunctions to variable degrees. Each patient showed at least one pathological result in the neuropsychological tests; vigilance impairment could be revealed only in 7 patients. P3-latencies were increased in OSAS patients when compared to age-matched controls (408.1 +/- 44.4 ms versus 373.4 +/- 32.5 ms; p < 0.03). P3 latencies and concentration on exertion showed a significant correlation with respect to the relative part of the total sleep time in which the patient's oxygen saturation was below 90% (p < 0.05). Thus it could be demonstrated that cognitive deficits in OSAS patients are a result of chronic intermittent oxyhaemoglobin desaturation rather than of a decline in daytime vigilance. ERPs represent an objective neurophysiological tool in neuropsychological examination. As they are also generated in subcortical cerebral structures they may indicate cognitive dysfunctions which cannot be evaluated by neuropsychological tests. If lead of ERPs is possible in special sleep centres they should be additionally used in the assessment of cognitive functions in OSAS patients. PMID- 9340625 TI - [Actigraphy: methodological limits for evaluation of sleep stages and sleep structure of healthy probands]. AB - Purpose of the investigation was to evaluate the differences of movement density during the sleep stages and waking. 22 diurnally active, healthy, male volunteers of mean age 30.7 (+/-Standard deviation +/- 3.3) years and a Body-Mass-Index 23.6 +/- 3.3 kg/m2 participated in the study. All subjects were recorded in the sleep lab via cardiorespiratory polysomnography and wrist actigraphy (Ambulatory Monitoring, Ardsley, USA) worn on the non-dominant hand, for two consecutive nights. The activity data, consisting of the number of zero crossings (NZC) were recorded in 1-minute periods. Sleep stages were scored visually according to standard criteria. EEG- and actigraphy data were converted to the same data format (European Feature Files). Attaching the actimetry data to the sleep stages was calculated mean NZC for every sleep stage and Wake. In spite of high differences in total individual NZC we observed that most NZC occurred during Wake. NREM 1 movement density was significantly higher in 19 recordings (86%) than in any other sleep stage. In 18 cases (82%) lowest movement density was found in NREM 3/4 with significant difference to all other sleep stages. Within 50% of the recordings were found decreasing activity in the following sequence of stages: Wake > NREM 1 > REM > NREM 2 > NREM 3/4 However, in all other cases there was a varying pattern of activity. CONCLUSION: Although there is some correlation between motor activity and sleep stages, the predictive value of actimetry data analysis in the assessment of sleep structure appeared to be limited mainly by individual movement density, especially during REM and NREM 2. PMID- 9340626 TI - [Pronounced hypersomnia in a 13-year-old patient with periodic leg movements]. AB - We report on a rare case of a thirteen-year old female with excessive daytime somnolence, which was caused by periodic leg movement. This underlines the necessity first to consider this diagnosis and second to record unconditionally the EMG derivation at minimum of the tibialis muscles in all patients with hypersomnolence. Even if carbamazepine is not the treatment of choice, in this case it was of remarkable effect. PMID- 9340628 TI - [Heart rate variability in patients with sleep associated breathing disorders]. AB - The increased mortality among patients with obstructive sleep apnoea syndrome has been explained in part by the increased incidence of pulmonary hypertension and coronary artery disease (CAD). A decreased heart rate variability has been shown to be associated with an increased mortality as well. We therefore screened 53 patients for sleep-related breathing disorders (SRBD) and heart rate variability (HRV) during the sleeping period. Standard time domain parameters were compared in a univariate multifactorial model for patients with an oxygen desaturation index (ODI) of more or less than 5 including the factors CAD, diabetes and beta blocker use. The percentage of differences between RR-intervals that differ more than 50 ms (pNN > 50: 9.0 +/- 11.1% vs 19.2 +/- 22.2%; p < 0.05) as well as the root mean square of these differences (rMSSD: 38.0 +/- 29.0 msec vs 59.2 +/- 51.5 msec; p < 0.05) were significantly decreased in patients with SRBD. These results favour HRV for inclusion in future risk stratification models in patients with sleep-related breathing disorders. PMID- 9340627 TI - [Sleep and respiration at an altitude of 6,400 m (Aconcagua, Argentina]. AB - Persons at extreme altitudes are known to experience disturbances in the regulation of ventilation and sleep structure. However, except for simulated studies using the decompression chamber, only single events of sleep or ventilation were measured so far in field studies up to an altitude of 5800 m. Modifying a portable sleep lab (Vitalog HMS 5000), we were able to conduct 7 channel polygraphy on our ascent to the Aconcagua up to an altitude of 6400 m. METHODS: In 6 climbers (age 38-62 y, 1 f, 6 m), ECG, EOG, SaO2, chest and abdominal movements, breathing and snoring sounds, body position, nasal and oral airflow were measured 4 weeks prior to the expedition at an altitude of 500 m, at base camp (4200 m) and in 3 climbers at 6400 m (2nd base camp) at the Aconcagua mountain. All participants had a repeat study at 500 m altitude 4 weeks after the expedition. RESULTS: The total number of obstructive apnoeas and hypopnoeas (OA/H) at night increased at an altitude of 4200 m in the mean of all 6 climbers from 36 to 67.7 compared to 500 m altitude, Central Apneas and Cheyne stokes (CA/CS) increased from 6.7 to 45.2. At 6400 m altitude the OA/H fell to 3 and 4 respectively in 2 climbers and CA to 1 and 2 respectively. In one climber, suffering from recurrent snoring with oxygen desaturation at 500 m altitude level, the number of OA/H and CA/CS increased further to 201 and 322, respectively, at 6400 m. Total sleep time including the REM position increased in all 6 climbers by 10% at base camp in comparison to an altitude of 500 m. Whereas the total sleep time remained constant in the 3 climbers at 6400 m altitude, the REM position declined by 10% in comparison to base camp (4200 m). However, significant fluctuations between individuals were noticed. CONCLUSION: Although significant alterations in sleep and breathing are noticeable at altitudes above 300 m, the respiratory drive in healthy subjects provides for a regular ventilation at high frequency at the extreme altitude above 6000 m. Sleep-related breathing disturbances at low altitude appear to be amplified at high altitudes. PMID- 9340629 TI - [Effect of long-term oxygen therapy on sleep architecture in patients with severe dilated cardiomyopathy and Cheyne-Stokes respiration]. AB - Patients suffering from severe heart failure may develop breathing disorders during sleep. Results may be heavy disturbances in sleep architecture, worsening of haemodynamics and of the prognosis of these patients. Causes of breathing disorders are probably instability of breathing regulation caused by hypoxaemia, hypocapnia, and prolonged blood circulation time. This study examined the influence exercised by different concentrations of continuously applied oxygen during night time on breathing disorders, oxygen saturation and sleep architecture in patients with severe heart failure (NYHA III-IV). All patients showed an improvement in sleep architecture. Total sleeping time increased significantly. Fragmentations of sleep by arousal reactions decreased, time of REM-sleep and non-REM-sleep III and IV increased significantly. PMID- 9340630 TI - [Carotid baroreflex in sleep apnea patients]. AB - 15 patients (11 male, 9 female) with obstructive sleep apnoea syndrome (OSAS) and 10 healthy age- and weight-matched control subjects volunteered to participate in the study to test circadian variations of cardiac response to activation of carotid baroreceptors. Activation of carotid baroreceptors was evoked by applying a negative pressure to both left and right carotid sinus regions, using two small separate neck-suction capsules. All experiments were performed under standard conditions every two hours. The cardiac response to carotid baroreceptors activation was significantly reduced in OSAS Patients, to a greater degree in the group of hypertensive patients. A characteristic circadian rhythm of baroreceptor reactivity persisted in all OSAS patients. Thus, OSAS patients constitute a high risk group for the development of cardiovascular diseases and pathological course of blood pressure during daytime. PMID- 9340631 TI - [Use of discontinuous long-term blood pressure measurement (Spacelabs 90207) in patients with sleep apnea--a comparison with intra-arterial data]. AB - Circadian blood pressure (BP) profile is an important determinant of cardiovascular risk. Patients with Sleep-Related Breathing Disorders (SRBD) often suffer from arterial hypertension and an altered circadian blood pressure profile. The aim of this study was to compare intraarterial blood pressure recordings with DBPR (Discontinuous Blood Pressure Recorder using Spacelabs 90207) in 20 patients with mild to moderate arterial hypertension and mild to moderate SRBD. Our results of overnight measurements show that the mean arterial pressure seems to be the most reliable value (mean difference Spacelabs-Part: +1.7 mmHg). In contrast, the systolic BP was systematically underestimated (mean: -17.5 mmHg) while the diastolic value was systematically overestimated (mean: +9.3 mmHg) by Spacelabs. Our data show that neither systolic nor diastolic BP from the Spacelabs measurement reflected the real cardiovascular load. Since the mean arterial pressure proved to be the most reliable value, this value should be used to distinguish between dipper and non-dipper in circadian BP profiles. Further, reliable noninvasive continuous measurement of the BP is required to assess the real vascular load in patients with SRBD during the night. PMID- 9340632 TI - [Increased daytime tiredness, nocturnal hypertension and sleep apnea: studies in rural general practice]. AB - Studies in a Rural General Practice: Sleep-Related Breathing Disorders (SRBD) and altered circadian blood pressure profile are related to increased cardiovascular risk. We investigated the prevalence and coincidence of both diseases in male patients from a general practice in a small community of 2500 people. Out of 409 selected patients (using a questionnaire regarding symptoms and findings of SRBD), 185 were monitored in an outpatient setting with an apnoea-screening system Mesam IV. Ambulatory blood pressure monitoring (Spacelabs 90207). Holter ECG and actigraphy were also measured. Sixty patients had a Respiratory Disturbance Index (RDI) > 10. An indication for further sleep studies was seen in 40 patients; 36.5% of daytime hypertensives were "Non-Dipper", and 47.6% of normotensives were also "Non-Dipper". Excessive daytime sleepiness (EDS) is an important symptom of SRBD, and in this investigation we noticed a large number of patients without this symptom suffering from relevant SRBD. Therefore, absence of EDS alone is not indicative for the use of ambulatory monitoring. PMID- 9340633 TI - [Computer-assisted nCPAP settings in comparison with conventional methods]. AB - CPAP with constant pressure is the method of choice to treat obstructive sleep apnoea. To find the optimal pressure, automatic systems were developed, permitting computerised titration of pressure based on respiratory patterns. One important point in the algorithm of titrating pressure is the shape of the inspiratory breath flow-time relation. It can be used to detect early inspiratory obstruction. In the present study the results of automatic CPAP titration were compared with conventional constant CPAP. In 12 patients with obstructive sleep apnoea syndrome two polysomnographies were performed after diagnosis and conventional titration. In a randomised cross-over study the adequate constant CPAP was compared with computerised variation of pressure by Autoset-CPAP. During both methods apnoeas, hypopnoeas and snoring were reduced. In the autoset mode the number of apnoeas and hypopnoeas was 30.14 +/- 17.6/sleeping time, in the constant mode it was 7.9 +/- 11.2/sleeping time (p < 0.01). The number of episodes with snoring in the autoset mode was 18.7 +/- 21.8 and in the constant pressure mode 7.9 +/- 11.2 (p = 0.054). No difference could be detected in respect of sleeping time, sleep efficiency, arousals and sleep stages. We conclude that in patients accepting constant high pressures constant CPAP remains the method of choice. Automated CPAP offers an alternative in the case of pressure intolerance beneath BiLevel-CPAP. PMID- 9340634 TI - [Comparative study of thoracic and abdominal effort, respiratory oscillatory impedance (ROI) and intrathoracic pressure in sleep apnea syndrome]. AB - Oesophageal pressure (Pes) and oronasal flow are necessary to describe upper airway obstruction in patients with obstructive sleep apnoea syndrome (OSAS), but Pes interferes with sleep. We developed a device applying an oscillating flow (20 Hz) through a nasal mask. An additional flow (2.6 l/min) is needed to reduce dead space and humidity. 24 patients (age 55.8 +/- 8.3 years, BMI 28.6 +/- 3.9, RDI 38.6 +/- 19.4, Raw 0.27 +/- 0.07 kPa/s/l) underwent polygraphy (oronasal flow, thoracic and abdominal effort, oxygen saturation, microphone, heart rate). Pes and oscillatory impedance (OI) were measured simultaneously. During snoring, hypopnoeas and apnoeas we compared Pes, OI and effort values for the detection of number and period of airway obstruction. The average Pes during habitual snoring was -3.2 +/- 0.8, during hypopnoeas -3.9 +/- 1.1 and during apnoeas -4.4 +/- 1.6 kPa. We found no significant difference in respect of the number and period of obstruction in patients with apnoeas, whereas in patients with incomplete obstruction (hypopnoea) Pes and OI were found to be more sensitive in detecting obstruction than effort (period: 27.0 +/- 9.1 sec (Pes), 29.0 +/- 4.8 sec (OI) vs. 20.0 +/- 6.8 sec (effort); number: 34.0 +/- 9.1 (Pes), 35.0 +/- 8.5 (OI) vs. 23.0 +/- 9.5 (effort). There is a significant correlation between Pes and OI (r = 0.89). OI is shown to be equally sensitive in identifying Upper Airway Resistance Syndrome as compared to Pes. This method is more convenient than conventional measurements such as Pes and it could be an alternative. PMID- 9340636 TI - [Change in bronchial hyperreactivity with nCPAP respiration in patients with sleep related respiratory disorders]. AB - Own results of 23,174 patients suffering from sleep apnoea syndrome showed that about 4 per cent of these patients have an increase of bronchial reactivity. In 60 patients with assured bronchial hyperreactivity we performed an inhalative bronchial provocation test using carbachol. Tests were performed before and after two days of treatment with nCPAP in the early morning. 23 of our 60 patients showed a decrease of PD20 FEV1 after nCPAP. The other patients did not show any significant differences. Our results demonstrate that in patients with increased bronchial reactivity, nCPAP therapy may aggravate bronchial hyperreactivity. PMID- 9340635 TI - [Incidence of cutaneous sensitization to environmental allergens in obstructive sleep apnea syndrome]. AB - A skin prick test was performed in 75 patients with an obstructive sleep apnea syndrome (OSAS) to test allergic skin reactivity to 18 common inhalant allergens. Additionally, anterior rhinomanometry was performed. Results were compared with a group of 21 patients with chronic obstructive pulmonary disease and with 198 workers of the chemical industry. A positive (> or = 4 mm wheal diameter) skin test to at least one allergene was present in 49 per cent of the OSAS patients, compared to 14 per cent in COPD patients and 28 per cent in industrial workers (p < 0.05; chi 2-test)-33 per cent (n = 25) of the OSAS patients with a sleep apnea syndrome showed skin reactivity to house dust mite (D. ph., D. f.) compared to 9.5 per cent in COPD patients and 12.6 per cent in industrial workers (p < 0.05 chi 2-test). No difference was found between anthropometric, polysomnographic, or rhinomanometric data comparing OSAS patients with positive skin reactions against house dust mites (n = 25) and those without any allergic skin reactivity (n = 38). PMID- 9340638 TI - [Effect of n-CPAP therapy on outcome of cold provocation]. AB - Our own investigations comprising 23,174 patients suffering from sleep apnoea showed that about 4 per cent of these patients suffer from a hyperreactive bronchial system. In some of these patients treatment with nCPAP causes coughing or mild dyspnoea even after having been previously asymptomatic. Loss of water and heat on the surface of bronchial mucosa may induce reversible bronchoconstriction. We examined in 60 patients suffering from obstructive sleep apnoea whether mechanical treatment with nCPAP would cause a change in bronchial reactivity. Cold air hyperventilation was used in provocation testing. Provocation tests were performed before and after a 3-day treatment with nCPAP in the early morning. In some patients with previously positive reaction, application of nCPAP alone decreased the lung function. Cold air hyperventilation challenge may be helpful to detect possible risks in patients using nCPAP, and to minimise such risks. PMID- 9340637 TI - [Bronchial hyperreactivity and nCPAP therapy]. AB - In patients with obstructive sleep apnoea (OSA) nCPAP may irritate the mucous membranes of the upper airways. We investigated in this study whether nCPAP can induce bronchial hyperreactivity (BHR). Forty-one patients (33 men, mean age 52.6 years) were treated with nCPAP due to OSA. All of them were tested for BHR with histamine ("pari-provo-Test") before and six weeks after initiation of the nCPAP therapy. Thirty-five of the patients showed BHR neither before nor after the beginning of CPAP. Six patients developed a BHR of moderate degree (PD20: 50-100 micrograms) during the study; four of these six patients were not symptomatic. The two other patients complained about more colds than usual or about noctumal cough. Both of them received inhaled steroids and a moistening system. Nobody of the enrolled patients was obliged to finish CPAP therapy due to BHR. Four patients had already a BHR before nCPAP therapy began. Most of the patients did not acquire a BHR during the first 6 weeks after nCPAP therapy had started. A BHR bronchial may develop, but in the majority it remains without clinical relevance. In patients with a BHR and OSA, the benefits of nCPAP therapy excel the potential adverse effects. PMID- 9340639 TI - [Do patients with obstructive sleep apnea syndrome treated with nCPAP therapy lose weight?]. AB - A simple therapy for obstructive sleep apnoea syndrome is weight reduction, which we always recommend before initiating and maintaining nCPAP-therapy. We documented the body weight of 123 patients (9 women, 114 men; age 53.8 +/- 10 years, initial apnoea-hypopnoea-index 40.7 +/- 22.6/hour) before and after 582 +/ 391 days nCPAP-therapy. Absolute and relative (Broca Index: weight [kg]/[height [cm] -100] x 100) body weight was 97.8 +/- 19.4 kg resp. 128.3 +/- 24.3 units before and 97.3 +/- 18.2 kg resp. 127.8 +/- 23.5 units during therapy (not significant). Body weight changes ranged from +22 to -26 kg. Only a subgroup of patients with a Broca index between 100-119 exhibited a significant weight change from 86.8 +/- 10.4 to 88.5 +/- 10.4 kg. We conclude that our recommendations to lose weight were unsuccessful in patients with obstructive sleep apnoea, although nCPAP-therapy generally improved well-being. PMID- 9340640 TI - [Physical training of patients with sleep apnea]. AB - PURPOSE: It is a common question of sleep apnoea patients in the sleep lab whether they stand a chance to decrease the symptoms and severity of their disease by physical exercise. As far as we know, there is no data about this specific question until now, even though this has been subject to speculation. A few studies, however, report on an improvement of the respiratory drive (and chemoreceptor sensitivity) after physical exercise in athletes. The aim of this study was to prove whether physical exercise in sleep apnoea patients could improve the symptoms of their disease in an open trial. METHODS: 11 Patients with mild to severe sleep apnoea syndrome (1 f, 10 m, mean age 53.8x) took part in a 6 month period of physical exercise twice a week 2 h each time under the instructions of physical therapists. Before and after the 6mo period a full PSG without CPAP or BIPAP, a bicycle exercise test with lactate profile, echocardiography, blood test, and body weight and body height measurement was performed. Statistical analysis was done using Wilcoxon ranked test and multiple regression analysis. RESULTS: There was no significant bodyweight reduction in all patients after the 6mo period of physical training, no significant difference in either basal SaO2 nor mean SaO2 and no significant improvement in physical status by the p at 4 mmol lactate on the lactate profile. Echocardiographic changes were not found; there was no significant change in the blood pressure profiles during the bicycle test. No cardiopulmonary problems including exercise induced high blood pressure were reported during the training period. There was, however, a significant decrease of the RDI (p < 0.05), but no significant change in the REM-sleep % of total sleep time (TST) and the TST itself. CONCLUSIONS: There was an improvement of the sleep apnoea syndrome correlated to a decrease of the RDI in the studied patient population due to a possible increase in the respiratory drive or a stabilised muscle tone ine the upper airways after physical exercise, as reported by other authors, because weight reduction could not be the reason in our patients. Our trial showed that the exercise does not increase the severity of symptoms of sleep apnoea by changing the REM/non REM ratio or for any other reasons. A physical training programme for sleep apnoea patients as an additional treatment should therefore be considered. PMID- 9340641 TI - [Effect of alcohol on minimal effective nCPAP pressure]. AB - Alcohol may have an aggravating effect on sleep disordered breathing. Aim of our study was to test the effect of alcohol on the required nCPAP pressure as determined by the self-adjusting nCPAP-system AutoSet. Ten male subjects (age 54 +/- 9 yrs, body mass index 37 +/- 5 kg/m2) with moderate to severe obstructive sleep apnoea (OSA) were investigated. Full polysomnography was performed on four consecutive days (control night with and without alcohol, nCPAP pressure determination by AutoSet with and without alcohol, in randomised order). Alcohol was given in a single dose of 80 proof vodka (1.5 ml per kg of body weight) one hour prior to bedtime. Alcohol to a deterioration of the respiratory disturbance index (RDI, 56 +/- 23 without vs. 66 +/- 19 with alcohol, p = 0.02), but no significant change was observed in mean or minimal oxygen desaturation, mean or maximal event duration. The 95th percentile of the AutoSet-pressure was not different with or without alcohol (10.7 +/- 2.5 vs. 10.6 +/- 2.5 cm H2O). Moderate alcohol intake in the evening need not be taken into account for CPAP pressure determination in moderate to severe OSA. PMID- 9340642 TI - [Extent of variations in minimal CPAP pressure requirements during a single night]. AB - The frequency of apnoeic events during sleep depends on posture and sleep stage. This is due to a change of upper airway size and upper airway closing pressures during different postures and sleep stages. The extent of the change of the necessary "splinting" pressure during nasal CPAP therapy for obstructive sleep apnoea is unknown. We titrated during CPAP therapy the minimal effective pressure, depending on sleep stage and posture. The necessary pressure fluctuated, depending on the patient, between 3 and 10 mbar. The highest pressure was not solely achieved during supine position and REM; some patients needed higher pressures under other circumstances. PMID- 9340643 TI - [Technical differences in various CPAP and BiLevel CPAP devices]. AB - PURPOSE: Since the first presentations of CPAP by Sullivan 1983 und BiPAP by Sanders 1990 as a successful treatment in obstructive sleep apnoea syndrome, many CPAP and BiLevel-CPAP devices have been developed by several companies around the world. Although all devices work on the same principle of continuous positive airway pressure delivered through nasal or facemasks, there are, however, significant technical differences between these devices, mainly due to different size and different maximum speed of the turbines. As far as we know, the only study concerning this technical difference in the devices was conducted by Raschke in 1995, who found significant differences in the pressure stability of the devices using a very complicated measurement model on volunteers who were breathing on the different devices. We intended to study the technical differences of the devices using a more simple technique in static conditions, but with very exact measurement. METHODS: We measured pressure stability on different inspiration flows and different pressure levels of 8 CPAP's and 4 BiLevel-CPAP's under static conditions using the "Hontzsch Exact ASD-G Messrohr ms 20201-18" flow-measurement device and the "Thommen HM 18.0020.A" pressure measurement devixe. We measured the noise emission by these devices at different frequency levels using the "Bruel and Kjer Dual Channel Real Time Frequency Analyzer", and measured the speed to reach the adjusted pressure level with the "Multimeter Phillips PM 2518X" and "Oszillograph Phillips PM3350" using the voltage change due to different working of the electric engines at different turbine speeds. We also investigated the different construction of the devices by opening them and analysing the materials used in the devices. RESULTS: The real pressure levels at an adjusted pressure of 10 mbar and at an inspiration flow of 1 Liter/sec range from 8.7 mbar to 15.6 mbar in CPAP's and from 9 mbar to 9.7 mbar in BiLevel devices. At higher inspiration flows the differences are larger, at lower flows smaller. The maximum noise emission in the 10 Hhz spectrum at a distance of 100 cm from the device at an adjusted pressure level of 10 mbar ranged from 29.6 dB to 39.9 dB in CPAP's and 30.4 dB to 42.2 dB in BiLevel CPAP's. The time to reach an adjusted pressure level of 10 mbar after closing the airway of the device ranged from 0.26 to 0.66 sec in CPAP's and from 0.46 to 0.80 sec in BiLevel devices. CONCLUSION: There are significant technical differences in the different CPAP and BiLevel devices due to different construction of the turbines and electrical engines of these devices. This should be considered when prescribing a device for a sleep apnoea patient. Not every device is suitable for every patient, and quality differences should be considered by all persons involved in the production and prescribing process. PMID- 9340644 TI - [Problems in nasal CPAP management of the obstructive apnea patient--when is a tracheotomy indicated? A case report]. AB - The use of nCPAP-ventilation has changed the rare indication for tracheotomy in patients with severe sleep apnoe syndrome. Only in cases where obstructive nasal and epipharyngeal processes are not resectable tracheotomy can be necessary. A case report shows one indication for such a surgical intervention. Increasing nasal obstruction usually can be treated successfully by operation. PMID- 9340645 TI - [The clinical use of an individually fitted nasal mask ("Freiburg Respiratory Mask") within the scope of a case report of controlled BiPAP ventilation]. AB - PURPOSE: Noninvasive mechanical ventilation with nasal or face mask using either BiPAP or IPPV (intermittent positive pressure ventilation) modes is meanwhile the standard type of mechanical ventilation instead of endotracheal intubation or tracheostomy in many patients with chronic and acute respiratory failure. However, problems occur very often in noninvasive mechanical ventilation due to mask problems with leakage of air out of the mask or erasions and necrosis of the skin by constant pressure through the mask frame. Hence, some clinical work groups developed customized molded masks mostly in cooperation with dentistry labs. These masks, however, are often very expensive and take several weeks to be manufactured. Our aim was to develop an inexpensive (less than 600 USS) and easy (in one day)-to-produce customized nasal mask, the so-called "Freiburg nasal mask". We wish to show by a case report the clinical efficacy of this customized mask in severe respiratory failure. CASE AND METHOD: A 52 y old women (BMI 19, nonsmoker) suffering from a severe hypercapnic respiratory failure (PCO2 over 100 mmHg) due to a severe kyphoscoliosis as result of a postpoliosyndrome and already on controlled nasal BiPAP with a standard respironics nasal mask, was referred to our clinic as an emergency case. After not succeeding to normalise blood gases and clinical status of the patient in mechanical ventilation on BiPAP mode and additional 2 lit/min of oxygen insufflation via the mask (patient was refusing invasive ventilation) for 12 days due to mask leakage and mask discomfort we made a mould of the patients face. Using this mould in a vacuum process the two EVA polymers Erkoflex and Erkodent were combined to build up the customized mask over the mould on the same day. In the first three days using the customized mask in the patient PCO2 decreased below 70 mmHg, on the 8th day after starting with the customized mask, blood gases almost normalised. Patient compliance in using the BiPAP device increased from 8 h a day to 16 h a day. CONCLUSION: This case shows that sometimes due to leakage of air in standard nasal masks noninvasive mechanical ventilation may fail in patients with severe respiratory failure. Therefore, in these patients customized moulded nasal or face masks should be used instead of standard masks. We think that the Freiburg nasal mask is an example of an easy to produce and nonexpensive customized mask and may be used in such patients, as this case report shows. PMID- 9340646 TI - [Effectiveness of an anti-snoring prosthesis in obstructive sleep apnea syndrome]. AB - Nasal CPAP is a well documented successful therapy for obstructive sleep apnoea syndrome (OSAS), but adverse effects are common and the compliance is 80 percent or less. We investigated the value of an oral device called "Snor Ex"). Eleven obese patients with severe OSAS were examined. Complete polysomnographic studies were performed while sleeping one night without therapy, with nCPAP and after an adaptation period with Snor Ex, respectively. The apopnoeahypopnoea index and the minimal SaO2 could be normalised for all patients using CPAP but only for three patients with Snor Ex. Nine patients tolerated nCPAP and Snor-Ex, respectively. We conclude that this oral device is a successful therapy for only few patients with OSAS. PMID- 9340647 TI - [Is SnorEx also ApneaEx? A study with a new intra-oral prosthesis as a form of therapy of obstructive sleep apnea syndrome]. AB - In recent years different kinds of dental devices have been advocated for treating sleep apnoea. In this study we report on our results with a kind of prosthesis ("Snor-Ex", DEPITA, 29336 Nienhagen) designed to relieve upper airway obstruction in sleep in pulling forward the tongue by a truss pad positioned in the posterior area of the tongue. We performed the study to test the effectiveness of the device in reducing the number of obstructive events. PATIENTS: 23 patients with OSA (22 male, age: 53.7 +/- 8.6 years, Body mass index: 31.1 +/- 3.9 kg/m2, Apnoea index: 33.5 +/- 18.4, Respiratory disturbance index: 45.6 +/- 19.7, mean apnoea duration: 20.4 +/- 4.4 sec) were included. STUDY DESIGN: Before the study was started, polysomnography was performed and the OSA associated symptoms/claims were standardised with the help of visual analogue scales (VAS). The prosthesis were made by the dental laboratory. Between the 28th and 42nd day after beginning with the study the patients had to come to the hospital for control. The effect of the therapy was documented only by a further polysomnography in patients who could sleep for at least 2.5 hours with the prosthesis. The effects of the device on changing OSA-associated symptoms and snoring were reevaluated by the above mentioned VAS. During the control the patients were divided into non-responders (NR) and responders (R) according to the results. RESULTS: The NR prevail in the study with 75% (17/23). They are characterised by inacceptable loco-regional side effects of the prosthesis, missing improvement of the state of daytime wellbeing and constant obstructive events. Only 25% of the patients are R. They locally tolerated the prosthesis, which is the precondition for long-term therapy. The severity of OSA diminished. Snoring also diminished significantly. CONCLUSION: According to our results the insufficient acceptance and the low effectivity of the SnorEx-prosthesis preclude large-scale indication for OSA patients. The prosthesis should not be prescribed without contacting a sleep lab. PMID- 9340648 TI - [Predictive value of laryngoscopy with reference to the severity of obstructive sleep apnea]. AB - The diagnosis of sleep-related breathing disorders is based primarily upon targeted history-taking and upon night-time polysomnography. Diagnostic measures should be completed by laryngopharyngoscopy (LPS). Frequently in cases of relevant obstructive sleep-related breathing disorders, a collapse of the pharyngeal lumen may be detected already in the waking state: either during spontaneous breathing or through adequate provocation (snoring manoeuvre). Furthermore, endoscopy of the upper airways will yield indirect clues as to nocturnal snoring and possible obstructive apnoeas. Although the predictive value of LPS regarding the degree of obstructive sleep-related breathing disorders seems low compared with polysomnography, it appears reasonable to perform it to exclude further relevant ENT findings. With LPS, local and regional obstacles of the kind of the intended positive pressure ventilation (narrowing of the nasal ducts, septum deviation and hyperplasia of the turbinates) are demonstrable which may warrant surgical correction. LPS, which has a low complication rate, facilitates interdisciplinary cooperation in the care for patients suffering from obstructive sleep-related breathing disorders. PMID- 9340649 TI - [Adeno-tonsillar hyperplasia and obstructive sleep related respiratory disorders]. AB - The most common cause of obstructive apnoeas in children is adenotonsillar hyperplasia. Possibilities of therapy are surgical intervention (adeniodectomy, tonsillectomy) or nasal CPAP. The decision on the most appropriate therapy must be individual. 26 children aged 2-5 years were investigated polysomnographically before therapy, after operation or with nCPAP-therapy. Polygraphy is a helpful aid in making up one's mind in favour of one of the possibilities of therapy, but also for controlling the therapeutic effect. PMID- 9340650 TI - The leukemic core binding factor beta-smooth muscle myosin heavy chain (CBF beta SMMHC) chimeric protein requires both CBF beta and myosin heavy chain domains for transformation of NIH 3T3 cells. PMID- 9340651 TI - 4th International Congress on Essential Fatty Acids and Eicosanoids. Edinburgh, Scotland, 20-24 July 1997. Abstracts. PMID- 9340652 TI - [Psychiatry, homelessness and other unpleasant topics]. PMID- 9340653 TI - [Psychiatric disorders in homeless persons--Anglo-American studies of epidemiology and management]. AB - The state of Anglo-American research on psychiatric disorders among homeless persons is summarised. Several authors reported on life-time prevalence (50-75%) and present prevalence rates (30-50%) of psychiatric disorders in this population. Alcohol-related and drug-related disorders were most frequent. Most studies, however, only included single and male subjects, while there is little information on homeless persons living with a partner or in a family. Up to now, no representative studies exist. Some recent investigations evaluated classic single case models versus case management approaches. Short-term programmes only yielded short-term effects, but linking psychiatric and social services seems to improve health service utilization by the homeless. PMID- 9340654 TI - [Homeless patients in the psychiatric clinic. Results of a 12-month study of the living conditions of psychiatric patients in an urban public health clinic]. AB - In a psychiatric hospital with 3174 admissions in a year, 31% of the mentally ill and addicted patients have no private home. These homeless patients are surviving in different ways: 1. homeless without any kind of housing; living in the streets for a long time with high risks of morbidity and mortality; 2. living in shelters or hostels; 3. residents of local homes; 4. patients still have their private homes but they are urged to leave in the near future or they are living in dangerous circumstances with high risks for health. Case-management by social workers and local political interventions are necessary to act against these high risk health problems of homeless patients in a psychiatric hospital. PMID- 9340655 TI - [Mother-child treatment in psychiatry. II: Personal experiences--treatment concepts and special problems]. AB - Characteristics of 43 inpatient treatments of mother and child as carried out up to now, are described diagnostically and under a descriptive statistics aspect. The procedure of the staff is described, and the significance of the proceedings for the development of the child is also referred too. Specific problems (mothers with infants, mothers with toddlers, pre- and postnatal psycho-pharmaceutical therapy) are highlighted by means of case reports, or cameos. PMID- 9340656 TI - [Termination of inpatient treatment as a crisis and developmental tasks]. AB - Especially in the inpatient treatment of adolescents the phase of termination is decisive, because important issues and developmental tasks are re-experienced by the young patient. Therefore a sufficient relief from symptoms cannot serve as the main criterium for termination. Instead the ego-development with all its aspects and the improvement of the capacity for emotional relationship is most important. The paper discusses some central dynamic factors of the termination phase. Most of these processes are also induced inside the therapeutic team thus leading also to a kind of separation crisis in the team. PMID- 9340657 TI - [Self determination of patients in social psychiatric practice. A medical ethics model and its practical application]. AB - To protect patients' autonomy, valid informed consent must be given prior to medical treatment. This includes disclosure of information, understanding, ability to decide freely, and competence/capacity of the patient. However, a psychiatric disorder may disturb the ability to make a choice of their own. These patients have lost their competence to give valid informed consent. In everyday practice of social psychiatry, it is of particular interest to have objective and valid criteria to identify the incompetent patient. These problems are discussed in a case report, and four criteria (understanding, reasoning, decision making and appreciation of the disorder) are presented to diagnose the patients' competence. PMID- 9340659 TI - [Sex offenders in legal treatment and their relatives: results of family therapy sessions]. AB - Members of the family of origin and spouses have been invited to take part in a cognitive-behavioral treatment programme for incarcerated sexual offenders to support the offender. Offenders arranged appointments with their spouses, mother, fathers and the therapist. A one-year follow-up reveals considerable change in denying and minimisation of responsibility, forcefullness and degree of sexual intrusiveness of both the offenders and the spouses. Both groups showed a better understanding for and a more positive attitude to the offenders' sexuality. Furthermore, both groups showed a significant increase in applying concepts of relapse prevention. PMID- 9340658 TI - [Consumption patterns and motivation for use of addictive drugs in schizophrenic patients]. AB - In particular studies conducted in the U.S. display a tendency for schizophrenics to combine hallucinogens and amphetamines, whereas other studies report on a combination of psychotropic substances with a similar range of action. Influencing negative symptoms is reported to be the motivation for consumption. A total of 222 patients with a schizophrenic disorder (F2) and addiction (F1) were examined. The main substance was alcohol (F10.1 or F10.2; 52.2%), followed by cannabis (F12; 25%), opiates (F11; 4.1%), sedatives or hypnotics (F13; 2.7%) and cocaine (F16; 0.5%). A multiple drug use (F19) is reported by 14% of them. The most frequent combination was alcohol and cannabis, whereas hallucinogens and amphetamines were only rarely combined. Actual multiple consumption was reported by 55% of the patients, while lifetime multiple consumption applied to 72%. The motivation seems to be an unspecified sedation of unpleasant affective symptoms of schizophrenia. The most frequently seen combinations do not correlate with the reports published in the literature. The great variations in motivation seem to mainly reflect the importance of the availability of the substance. PMID- 9340660 TI - [Phototherapy and lithium prophylaxis]. AB - A random sample of patients (n = 43) had been investigated to detect any difference in response of bright light therapy in lithium-treated inpatients (n = 18) suffering from depression. Only 27% of the lithium-based inpatients respond to bright light therapy, but 73% of patients respond who were not treated with lithium. We conclude that lithium therapy reduces the chance of achieving t remission of depression by bright light therapy. PMID- 9340661 TI - [Organically-induced delusional syndrome in psittacosis]. AB - We report on a case of psittacosis which presented first as an atypical pneumonia and in the further course as a paranoid syndrome with a concomitant disorder of consciousness. In contrast with reports from the literature there were no obvious neurologic signs and symptoms which would have indicated the organic nature of the illness. Therefore, the correct diagnosis was based on a careful anamnesis, clinical and psychopathological examination and confirmatory results of serological tests. PMID- 9340662 TI - [What is psychiatry? Psychiatry in German language textbooks--2]. PMID- 9340663 TI - [Harm reduction--truly a paradigm change? Some sociological comments on the contribution by Thomas Kuhlmann]. PMID- 9340664 TI - [Disintegrative psychosis with main symptoms of dysfunctional social behavior]. PMID- 9340665 TI - [Therapy of affect incontinence with paroxetine]. PMID- 9340666 TI - [Urinary retention with risperidone]. PMID- 9340667 TI - [Comment on W. Hafner-Ranabauer et al.: Quality assurance in clinical social work: how do social workers assess the success of their interventions?]. PMID- 9340668 TI - [Self-awareness of a social worker or recognition determines awareness]. PMID- 9340669 TI - [Acute abdomen in childhood and adolescence]. PMID- 9340670 TI - [Necrotizing enterocolitis (NEC)]. AB - Necrotizing enterocolitis is a serious abdominal disease in infants during the first 2 months of life. The earliest radiographic finding on frontal abdominal films is initially distension of the small bowel, secondly of the colon, and pneumatosis intestinalis. Pneumatosis intestinalis is not pathognomonic for necrotizing enterocolitis; it can occur throughout life in different abdominal diseases. It can be a serious prognostic sign for abdominal ileus, but it may also occur in non-critically ill patients. Radiographic diagnosis in childhood is achieved with a frontal film of the abdomen in the prone position, and to detect a possible perforation, a left lateral abdominal decubitus exposure is necessary. Sonography and duplex sonography are helpful in evaluating progressive changes, the clinical course and the differential diagnosis. PMID- 9340671 TI - [Intestinal emergencies in newborn infants]. AB - Imaging plays a major role in most neonatal gastrointestinal emergencies. The role may vary from helping to establish a diagnosis, to the evaluation of associated abnormalities, to surgical planning, or to therapy for some conditions like meconium ileus or meconium plug syndrome. Plain radiographs and ultrasound serve a primary imaging modalities with bowel contrast examinations, CT scan, and MR imaging playing roles in more complex cases. PMID- 9340672 TI - [Volvulus in childhood]. AB - Gastric volvulus or volvulus of the small-bowel can occasionally be found in neonates and small infants. Since volvulus is an emergency case, the radiologist must know the characteristic radiological findings and the ultrasound signs in correlation to the clinical symptoms. Two forms of gastric volvulus can be distinguished: the organoaxial type and a mesenterioaxial form. Besides an idiopathic etiology, diaphragmatic alterations can be observed in children with volvulus of the stomach. Volvulus of the small-bowel occurs in children with malrotation type I or II or with nonrotation. Bile-stained vomiting starts within the first days of life and is followed by the clinical signs of high bowel obstruction and peritonitis. Primarily in cases of gastric volvulus, an ultrasound examination can show the wrong position of the stomach or the pyloric region. In cases of small-bowel volvulus, abnormal localization of the superior mesenteric artery can be demonstrated. The plain film features an upper small bowel obstruction. Upper intestinal contrast studies may reveal the level of small-intestine obstruction. A contrast enema can rule out a concomitant colon nonrotation or malrotation. A rare form which can be misdiagnosed easily, is volvulus of the sigmoid with pathological elongation and positioning of the sigma. It appears mostly in school children with less urgent symptoms and can disappear spontaneously. A typical feature is pain in the left lower abdomen and complete obstruction in an opaque enema. PMID- 9340673 TI - [Invagination]. AB - Intussusception is the most common abdominal emergency in infancy and childhood. Most cases are idiopathic ileocolic intussusceptions; rarely is a lead point present. Abdominal ultrasound is the imaging modality of choice for the demonstration as well as the exclusion of an intussusception. The target (transverse section) and pseudokidney (longitudinal section) signs are pathognomonic sonographic findings. Simultaneous depiction of lead points or lymph nodes or the presence of an entero-enteral intussusception may lead to different appearances. When an intussusception has been diagnosed with ultrasound, further complications such as small bowel obstruction or free intraperitoneal fluid have to be excluded at the same time. In addition, the perfusion of the intussusceptum can be evaluated with color Doppler ultrasound. There is general consensus that the only contraindications for conservative reduction are bowel perforation, peritonitis and hypovolemic shock. The oldest and most widespread method is hydrostatic reduction with barium under fluoroscopic control. Pneumatic reduction under fluoroscopic monitoring has gained more and more acceptance. An alternative technique is sonographically guided hydrostatic reduction with normal saline solution. Both latter methods are reported to have success rates of 80-90% and are clearly superior to the barium technique. In our opinion ultrasound monitoring offers the most precise control of the whole reduction process, with distinct demonstration of the intraluminal structures, especially of the ileocecal valve and of a possible lead point. A complication can be recognized immediately. The primary advantage is the lack of radiation exposure. Therefore, with appropriate equipment and experience this method may be regarded as most promising in the management of intussusception in infancy and childhood. PMID- 9340674 TI - [Appendicitis in childhood]. AB - From 1989 to 1995 high-resolution ultrasonography (US) was performed in 3,546 children (age: 1-17 years) with clinically suspected appendicitis. A total of 518 patients underwent laparotomy; 420 had histologically proven acute or perforated appendicitis (prevalence 11.8%). In these children, the sensitivity, specificity and overall accuracy of US examination were 90%, 97% and 96% respectively. The use of US in clinically doubtful acute abdomen may allow earlier diagnosis of acute appendicitis; in 1995 the rate of unnecessary appendectomy was reduced to 13%. PMID- 9340675 TI - [Gynecologic origin of acute abdomen in childhood]. AB - In cases of acute abdominal pain in girls a gynecological cause must always be considered. Neoplasms and cystic adnexal lesions complicated by hemorrhage, torsion, and infarction can be diagnosed in childhood. Ovarian tumors without endocrine activity are frequently very large at the time of discovery. Intravaginal foreign bodies, inflammation, and congenital obstructive malformations are seldom found in patients with acute abdominal pain. Ultrasound has become the diagnostic method of choice. When used by an experienced examiner, its results are nearly as good as MRI. In evaluation with clinical data and serological results, an exact diagnosis can be made by ultrasound, even if the sonomorphological pattern seems confusing. Solid adnexal mass and complex malformations require MRI as a complementary diagnostic step. X-ray studies and computed tomography are less important diagnostic tools in girls with acute abdominal pain caused by gynecological disease. PMID- 9340676 TI - [Dosage values for characterization of patient exposure in computerized tomography and their significance for risk assessment]. AB - In order to evaluate patients' exposure to radiation in computed tomography, dose quantities such as the computed tomography dose index (CTDI), multiple scan average dose (MSAD) and dose free in air on the axis of rotation are used. The CTDI and the MSAD, derivable from the CTDI, are a good measure for the absorbed doses in the examined volume, but they do not take the biological sensitivity of the organs into account and do not describe the radiogenic risk that is actually relevant for patients and doctors. The dose on the axis of rotation is not an accurate measure of the effective dose and the radiogenic risk. The declaration of a limit for the dose on the axis of rotation should take the different regions into account, in order to guarantee acceptable image quality on one hand and to avoid an unnecessarily high risk on the other side. As physical dose quantities, the CTDI, MSAD and dose on the axis of rotation are useful to characterize and differentiate between programs and systems concerning radiation exposure. PMID- 9340677 TI - [Methodological approaches to quantitative evaluation of microcirculation in tissues with dynamic magnetic resonance tomography]. AB - The development of rapid magnetic resonance imaging (MRI) sequences makes it possible to detect the fast kinetics of tissue response after intravenous administration of paramagnetic contrast media (CM), reflecting the status of tissue microcirculation. In this paper, the basic physical and tracer kinetic principles of dynamic relativity and susceptibility contrast MRI techniques are reviewed. The quantitative analysis of the acquired dynamic image data is broken up into an MR specific part, in which the observed signal variations are related to the CM concentration in the tissue, and an MR independent part, in which the computed concentration-time-courses are analyzed by tracer kinetic modeling. The purpose of the applied models is to describe the underlying physiological processes in mathematical terms and thus to enable the estimation of tissue specific parameters from measured dynamic image series. Whereas the capillary permeability can be estimated from dynamic relativity contrast enhanced MRI studies, the regional blood volume as well as the regional blood flow can be determined from dynamic susceptibility contrast enhanced image series. However, since there are no intravascular but only diffusible CM available at present, the application of the susceptibility technique is currently restricted to brain tissues with intact blood brain barrier. The practical realization of both dynamic MRI techniques is demonstrated by case studies. PMID- 9340678 TI - [Multiple, disseminated hypodensities in cranial CT. Multiple disseminated ischemia in pneumococcus-induced cerebral vasculitis]. PMID- 9340680 TI - [Unilateral lung transradiency. Swyer-James or MacLeod syndrome]. PMID- 9340679 TI - [Routine thoracic image of an 86-year-old patient with tracheobronchitis. Scalloping--pressure-induced ventral vertebral erosions caused by a thoracic aortic aneurysm]. PMID- 9340681 TI - [Contrast medium magnetic resonance angiography: a revolution in vascular diagnosis?]. PMID- 9340682 TI - [Invasive or noninvasive angiography? The role of "classical" catheter angiography]. AB - Within the last decade, diagnostic and interventional angiography have been developed to a high degree of performance, due to the widespread use od DSA, the miniaturisation of the puncture trauma and the introduction sets (catheters, sheaths), the development of high-tech materials (e.g., Nitinol guidewires) and the application of non-ionic, low osmolal contrast media. The specific risks of the procedure, thereby, have been significantly reduced, but could not be totally eliminated. To evaluate vascular diseases non-invasively, special attention was attributed to the progress of colour coded duplex, (spiral) CT-angiography and (CE) MR-angiography, based on the established imaging with US, CT and MRT. The matter is question is whether or not they can substitute the role of conventional angiography as the "gold standard" of vessel imaging. Clinical validity and economic efficiency both determine the indication for the use of invasive or non invasive methods. In diagnostic procedures, there is a growing tendency for the use of non-invasive techniques, like in imaging of the abdominal and thoracic aorta, the renal, pulmonary and extra- and intra-cranial arteries. Conventional angiography is still reserved for the evaluation of small vessels of the upper and lower extremities, and vessel status in preoperative conditions (carotid, celiac trunk, mesenteric and renal arteries and aneurysms of the cerebral vasculature). Fluoroscopic guiding of catheters and contrast enhancement in interventional procedures, however, cannot be substituted by alternative techniques in the foreseeable future. PMID- 9340683 TI - [Contrast medium enhanced magnetic resonance angiography. Theory, technique and practical implementation]. AB - Contrast enhanced (CE) magnetic resonance angiography affords angiographic depiction of extended vascular territories with high quality and diagnostic value. A prerequisite is the fast acquisition of a three-dimensional gradient echo data set during the injection of a bolus of a T1-shortening contrast agent. We describe the dependence of the quality of CE-MRA on technical parameters of different MR-scanners and consider some fundamental facts and practical guidelines concerning the contrast agent injection. PMID- 9340684 TI - [Indications for contrast medium administration in MR-angiography of cerebral blood vessels]. AB - Time-of-flight MR-angiography of large volumes is limited by the occurrence of saturation effects, which lead to low signal in both veins and arteries. Alternatively to a number of other MRA-techniques, intravenous application of paramagnetic contrast media in combination with 3D-pulse sequences with or without flow refocussing allows the depiction of slow vessels in large volumes without technical extra expenses. The main intracranial indication is anatomical 3D-imaging of normal and dysplastic cerebral veins with high spatial resolution, and the additional depiction of the venous drainage in AVMs, when unenhanced MRA shows only the arteriel supply and the nidus. In large cerebral aneurysms, bridging veins and venous sinuses, partial thromboses can easily be differentiated from slow flow. Contrast-enhancing tumors can be depicted together with normal or displaced vessels. Improvements of arterial signal due to contrast media with currently used routine MRA techniques are clinically not significant. Signal loss due to spin dephasing in vessels with complex flow is not influences by contrast media. Results of contrast-enhanced MRA are determined by the timing of injection. Since arteries and veins are both imaged with high signal intensity, improvements of postprocessing procedures for secondary vessel segmentation are necessary. PMID- 9340685 TI - [Conventional magnetic resonance angiography and contrast enhanced magnetic resonance angiography of extracranial blood vessel segments]. AB - The introduction of fast gradient systems allows a reliable visualization of the extracranial carotid vessels by the magnetic resonance angiography (MRA) which meanwhile is implemented into clinical routine. By the mainly applied time-of flight (TOF) technique, vessels can be imaged without contrast agent (CA). Due to the application of ultra-fast gradient-echo-sequences, the first-pass evaluation of an intravenous bolus-injection of Gadolinium in the carotids from the aortic arch up to the skull base can be performed in less than 30 s. In this study. Advantages and disadvantages of both techniques are discussed. For a qualitatively optimal contrast enhanced MRA (CE-MRA) timing parameters like injection delay, flow rate and the adjustment of sequence parameters have to be considered in relation to the fast venous return from the sinus to the jugular veins. First, the optimal time point of the data acquisition have been determined at a model and with a computer simulation in reference to the presence of CA in the arteries. As a result, 90% of the contrast contribution is defined by 16% of the symmetrically acquired central k-space lines. A measuring protocol for clinical use was obtained by a gradual variation of spacial resolution, measuring time and CA-injection parameters and was proved in normal volunteers and patients. An exact determination of the bolus-arrival-time by means of a test bolus injection was acquired. The best qualitative results were achieved by a double-dose- injection at 2 ml/s injection rate. The temporal reserves of ultra fast sequences should be invested in the improvement of the spatial resolution. To date, further investigations related to the problem of optimal CA-application may improve the potentials of CE-MRA procedures. PMID- 9340686 TI - [MR angiography of thoracic blood vessels]. AB - Through the introduction of newly invented high-performance gradient systems to MRI, which enable for echoplanar imaging (EPI), also magnetic resonance angiography (MRA) has gained an entirely new field of applications and techniques. Ultrafast imaging techniques in MRA allow the investigation of larger vascular areas within a single breath-hold-period. Artifacts like motion induced signal misregistrations, dephasing or saturation of the vascular signal are minimized by extremely short echo times. The technique thus requires the intravenous application of a contrast media bolus, usually a gadolinium compound, which is in standard clinical use. Coordination of the bolus injection and the timing of the data acquisition is crucial for optimal results. The first pass evaluation of the contrast media resembles CTA to a certain extend. Due to the fast measurement and the high contrast in contrast-enhanced MRA (CE-MRA) new applications and indications are developed like MRA of the pulmonary vessels. The paper offers considerations and trials for optimization of thoracical CE-MRA. Besides parameter constellation also bolus-optimization is described with respect to the dedicated anatomical premises. Investigations on volunteers and on patients build a basis for suggestions of optimized CE-MRA procedures. To date, a final estimation of the clinical value of the new technique cannot be given since ongoing improvements change the optimal protocol frequently and the potential of further developments is high. PMID- 9340687 TI - [Gadolinium-enhanced magnetic resonance angiography of abdominal blood vessels]. AB - Contrast enhanced (CE) magnetic resonance angiography (MRA) provides high resolution angiograms within 20-40 sec. The technique is based on the acquisition of heavily T1-weighted three-dimensional (3D) gradient-echo data sets (FISP) with ultrashort echo-(< 2ms) and repetition times (< 5 ms) during arterial phase of an intravenously injected bolus of a T1-shortening agent such as Gd-DTPA. For MR angiography of abdominal vessels CE-MRA is better suited than "time-of flight" (TOF) and phase-contrast (PC) MRA because motional artifacts can be obviated with breath-held acquisitions. We have optimised the technique and evaluated its potential for angiography of the abdominal aorta and its branches as well as the portal vein and its tributaries. Whilst CE-MRA provides reliable diagnostic accuracy in the aorta and the proximal sections of its branches, small peripheral arteries cannot be assessed accurately. The portal vein and its tributaries can often be depicted better with CE-MRA than with conventional angiography but, like conventional angiography, CE-MRA is hampered by slow and reversed flow, conditions under which TOF or "true FISP" MRA may perform bst. We have also investigated FLASH-echo-planar imaging (EPI) hybrid techniques, a further technical development which due to shorter acquisition times of 12-15 sec. allows semi-dynamic imaging of the arterial and venous phase and provide better vessel contrast due to the use of fat-suppression. PMID- 9340688 TI - [Value of contrast-enhanced 3D magnetic resonance angiography of the renal arteries]. AB - PURPOSE: To determine the value of gadolinium-enhanced, three-dimensional breath hold Magnetic Resonance Angiography (MRA) in the assessment of the aorta and renal arteries in comparison to conventional arteriography (CA). PATIENTS AND METHODS: 49 patients were evaluated with both CA and 3D MRA. 0.3 mmol/kg BW gadolinium-DTPA was administered intravenously in a bolus, using an automated injector. A test bolus method was used for timing of the bolus and beginning of the data acquisition. The intraarterial CA was used as the gold standard. RESULTS: MRA-based assessment of renal artery stenosis was identical with CA in 31 of 45 stenoses (68.8%). Sensitivity and specificity for assessment of renal arterial disease by MRA were 84% and 96%; for clinically relevant lesions they amounted to 90% and 98%. CONCLUSION: The presented contrast-enhanced 3D MRA technique allows for the reliable assessment of renal arterial morphology and pathology. PMID- 9340689 TI - [MR angiography: abdominal veins]. AB - Conventional MR-angiography (MRA) of the abdominal veins is a noninvasive and useful technique that has been accepted in clinical routine. The aim of this presentation is to evaluate the actual status of MRA including contrast-enhanced techniques and to compare clinical results with alternative diagnostic procedures such as ultrasonography, CT, and digital subtraction angiography (DSA). MATERIAL AND METHODS: Four different MRA techniques available today are presented that can be used for examination of the abdominal veins including sequence-parameters of the 2D TOF-FLASH technique, the 2D/ 3D PC techniques, the 2D True FISP technique, and the ultrafast 3D FLASH technique following i.v. administration of Gd-DTPA. RESULTS: From data of the current literature usefulness and accuracy of MRA is demonstrated in different provinces of the abdominal veins. DISCUSSION: On the basis of specific questions that are focused on MRA of the abdominal veins the value of this method is discussed under consideration of different techniques. MRA is evaluated under consideration of critical vascular areas, the role of contrast-enhanced MRA, suitable doses of contrast material, and new hard-ware configurations in the future. Furthermore, advantages and drawbacks of conventional MRA are compared with contrast-enhanced MRA-techniques, ultrasonography, CT and DSA. PMID- 9340690 TI - [MR angiography of pelvic arteries]. AB - During the last years, magnetic resonance angiography (MRA) has become a widely used modality for intracerebral and carotid artery imaging. Due to technical limitations, the clinical impact of MRA in the iliofemoral arteries has been rather poor. New developments in MRA like ECG-triggered sequences and the occurrence of contrast-enhanced MRA has overcome most of these limitations. Therefore, a major advance in clinical use of these diagnostic tools can be predicted. This paper discussed the advantages of ECG-gated 2D-Phase contrast, ECG-gated 2D-Time-of-Flight and contrast enhanced FLASH 3D angiography sequences from a clinical point of view. 2D-PC-MRA is a robust technique, which provides an overview of the iliofemoral artery system in less than 5 minutes. Limitations are the true 2D impression of the sequence and the partial venous overlay. 2D-TOF-MRA on the other hand is time consuming, however it enables 3D reconstruction and effective venous suppression can be applied. Contrast enhanced MRA as the third sequence discussed provides high resolution images in less than 30 sec. However contrast bolus timing might be a problem. In conclusion the authors suggest a combination of 2D-PC-MRA and additional 2D-TOF sequences at questionable vascular areas as the modality of choice, due to the fact, that MRA of the iliofemoral arteries ist mostly only one step of a complete lower limb examination. Contrast MRA might become the method of choice in the future however problems with multiple contrast injections and upper limits of contrast dose have to be solved. PMID- 9340691 TI - [Phase-contrast MR angiography of the lower extremity. Comparison of methods and clinical application]. AB - PURPOSE: To investigate whether phase-contrast MRA is a clinically suited approach to examine arteries of the pelvis and lower extremities. METHODS: The study was divided into two parts, a volunteer study and patient study. Three MRA techniques-2D TOF with venous saturation, 3D magnitude contrast and 2D phase contrast with ECG triggering-were intraindividually compared in 15 volunteers and evaluated by three blinded readers. Subsequently, a total of 230 vessel segments of 45 MRA studies using ECG-triggered phase contrast were compared with intraarterial DSA. All vessel segments were scored by three blinded readers using a five-point scale with DSA serving as the gold standard. RESULTS: ECG-triggered phase contrast provided better image quality than the other MRA techniques as assessed by the Friedman test. Clinical studies demonstrated a significant correlation of DSA and MRA as assessed by the Spearman correlation and kappa statistics for individual readers. CONCLUSION: MRA of the pelvis and lower extremities may be performed with 2D ECG-triggered phase-contrast MRA within a reasonable time frame (< 30 min). MRA slabs provide orientation similar to that with DSA projections and good to very good correlation of vessel pathology as shown by kappa statistics. PMID- 9340692 TI - [Contrast medium enhanced MR angiography of peripheral blood vessels]. AB - Currently 2D time-of-flight MR angiography (MRA) is most commonly used for the evaluation of the extremities. The major limitation is their susceptibility to signal loss from intravoxel phase dispersion. This leads to overestimation in the grading of stenoses. Further difficulties are motion artifacts and the limited spatial resolution. Phase-contrast MRA is increasingly used. However, further studies are required to verify its diagnostic potential. MRA technology continues to advance rapidly. Stronger gradient fields, shorter echo times and high resolution surface coils are developed. Contrast medium can be applied under standardized conditions with MR compatible power injectors. In phantom studies, the contrast-enhanced gradient echo sequences show reduced phase dispersion, resulting in an improved grading of stenoses. Initial patient studies underline these findings. Contrast-enhanced MR angiography has the potential to replace invasive arteriography in the evaluation of peripheral arterial vessels in the near future. PMID- 9340694 TI - [Arrhythmogenic dysplasia of the right ventricle]. AB - Arrhythmogenic right ventricular dysplasia is a disease of the cardiac muscle of unknown etiology. Landmarks of this disease are the presence of muscular atrophy and replacement of ventricular myocardium by adipous and fibroadipous tissue. This disease was originally described by Fontaine et al in 1977 during surgical ablation of drug refractory ventricular tachycardias in patients without evident structural heart disease. During surgery anomalies in contractility of the right ventricle and the presence of adipous tissue were documented. Some years later, Markus et al reported the first clinical series of patients with arrhythmogenic right ventricular dysplasia. Since then, this disease has been widely recognized and must be considered in the differential diagnosis of all patients with ventricular arrhythmias originating in the right ventricle. PMID- 9340693 TI - [A rare differential diagnosis of suspicious contrast medium enhancement in breast MRI. Melanoma metastasis in both breasts]. PMID- 9340695 TI - [Familial auricular fibrillation]. AB - INTRODUCTION AND OBJECTIVE: We present two families with atrial fibrillation in 20 of 50 members, during three generations, with known cardiac rhythms, in order to communicate their infrequent existence and the most relevant clinical facts. METHOD: Clinical situation, evolution, ECG and ECHO findings, treatments and complications related with the disease are investigated. RESULTS: The presence of atrial fibrillation in 20 members is demonstrated, although one of them was on sinus rhythm at the time of the study; 3 patients had left ventricular enlargement on the ECHO study; the clinical situation was good in all patients except two who died because of complications related to the arrythmia and a third patient that had a brain stroke. The patients received different treatments because they where controlled by different physicians; the possible lethal proarrythmic effect in such cases must be taken into account. CONCLUSION: Familiar atrial fibrillation is a very infrequent arrythmia, usually well tolerated, that follows a dominant autosomic hereditary pattern. The use of antiagregants is advised because of the risk of embolism, or the use of anticoagulants in the presence of associated risk factors. Electric cardioversion has been show not be useful. The possible proarrythmic effect of some antiarrythmic agents, used in the control of cardiac frequency, must be taken into account. PMID- 9340696 TI - [Evaluation of biochemical markers of myocardial lesion after radiofrequency ablation. Value of troponin I]. AB - INTRODUCTION AND OBJECTIVES: Radiofrequency catheter ablation is the curative treatment of choice for many cardiac arrhythmias. After radiofrequency ablation there is always a localized endomyocardial necrosis, which is necessary to eliminate the arrhythmia. The volume of the necrosis may be evaluated by the rise of several biochemical markers, classically CK and CK-MB. However, the sensitivity and specificity of these markers are not optimal and are probably less than ideal for this purpose. Cardiac Troponin I (cTnI) is a newly available biochemical marker available, with a high cardiac specificity. We designed this study in order to determine the value of serum levels of several cardiac markers in patients who underwent radiofrequency catheter ablation and to establish the utility of cTnI. METHODS: We analyzed the data from 51 patients who underwent radiofrequency ablation and from 16 control patients. In respect to the ablation target, we included in the study 14 left accessory pathways, 7 right accessory pathways, 12 atrioventricular nodal reentry tachycardia, 5 ventricular tachycardia and 13 atrial flutter or fibrillation. The levels of CK, CK-MB, cTnI, and myoglobin were compared with clinical findings, ST-T wave abnormalities and the presence of arrhythmias after ablation. To evaluate the diagnostic capability for each biochemical marker we used the ROC curves. RESULTS: A pathological value of cTnI was found in 47 out of 51 (92%) patients in the ablation group. CK-MB had a lower sensitivity (63%). The sensitivity for the other biochemical markers ranged from 30% for CK to 67% for Myoglobin. The area under the ROC curve for cTnI was 0.9375, significantly superior to the other biochemical markers (0.86, 0.76, 0.75 for MB, Myoglobin, CK respectively) (p < 0.05). The lowest cTnI released was found in patients after nodal reentry tachycardia ablation and the highest after atrial flutter ablation. cTnI increased above normal values in 4 patients in the control group (patients who underwent an electrophysiological study without catheter ablation). We found a moderate level of correlation between the number of radiofrequency pulses and cardiac cTnI release (r = 0.69; p < 0.0001). The correlation was different in each target, ranging between r = 0.25 (p = NS, 0.43) for atrial flutter and fibrillation to r = 0.99 (p < 0.0001) for ventricular tachycardia. CONCLUSIONS: cTnI had the greatest sensitivity (92%) for detecting minor myocardial damage. Thus, we can conclude that the serum level of cTnI detects the minor myocardial damage produced by radiofrequency ablation. PMID- 9340697 TI - [Inflammatory response in acute myocardial infarction. Predictive values]. AB - INTRODUCTION AND OBJECTIVES: Our purpose was to investigate the significance of inflammatory acute phase response early after myocardial infarction. We also observed how these indices were influenced by trombolytic therapy. METHOD: We examined the blood samples of 200 non consecutive patients at the first day of acute myocardial infarction (155 [77%] males; mean age 65 +/- 13 years) to characterize the proteins and proinflamatory reactants profile. Results were correlated with hospital mortality. Thrombolytic therapy was administrated to 117 patients on admission and in these patients the samples were taken after the procedure. RESULTS: Overall mortality was 8%. Serum C-reactive protein (69 vs 41 mg/l), haptoglobine (237 vs 190 mg/dl), gammaglobuline (0.93 vs 0.84 g/dl), alpha 1-globuline (0.28 vs 0.23 g/dl) and alpha-2-globuline (0.7 vs 0.6 g/dl) were significantly higher in patients without trombolytic therapy. Conversely, patients who had received lytic therapy, had higher plasma concentrations of interleukin-1 beta (104 vs 40 pg/dl). The only clinical variable which was associated with mortality was a Killip class > or = 2 on admission (mortality = 21%; odds ratio = 5.2; p = 0.02). Other biochemical variables associated with a higher mortality were a white blood cell count > 10/nl (mortality = 12%; odds ratio = 5.4; p = 0.01), increased activated neutrophils > 80% (mortality = 18%; odds ratio = 5.4; p = 0.004) and C-reactive protein > 20 mg/l (mortality = 11%; odds ratio = 6; p = 0.05). Only patients with activated neutrophils > 80% on admission had a higher probability of dying during hospital stay (Exp[B] = 3.6; B = 1.2; r = 0.29; p = 0.001). CONCLUSION: The acute phase reaction in early myocardial infarction is determined by thrombolytic treatment. A high increase of activated neutrophils on patient admission is the only biochemical predictive value for hospital mortality. PMID- 9340698 TI - [Nifedipine in the treatment of moderate and severe arterial hypertension. Long term effect on arterial pressure and on the left ventricle]. AB - INTRODUCTION AND OBJECTIVES: Moderate-to-severe hypertension is less prevalent than mild hypertension, but it is responsible for more incidences of complications. Its complex treatment requires several drugs, and is often inadequate. This study assessed the efficacy and safety of nifedipine GITS (oral release osmotic system) as monotherapy, in addition to the effects on left ventricular hypertrophy, after a long term follow-up (one year). PATIENTS AND METHODS: Thirty patients with diastolic blood pressure above 105 mmHg were studied after a short placebo phase. They received nifedipine GITS as monotherapy in a single daily dose of 30 mg; dose titration was made the first three months according to response and until they reached figures equal to or below 95 mmHg. By M-Mode echocardiogram, left ventricular mass index and systolic function were calculated at the end of the placebo phase and at 3, 6, 9 and 12 months. Hematological parameters, lipid profile, electrolytes and liver enzymes were assessed at the same periods. RESULTS: In 70% of the patients the blood pressure reached values of 140-90 mmHg. In 16 patients with adequate M-mode recordings, a 12% reduction in left ventricular mass was observed without modification in systolic function. Five patients were retired: two due to adverse events and three due to different reasons (drop out, evidence for secondary hypertension). There were no changes of clinical significance in the hematological or biochemical parameters and no hypertensive crisis occurred. CONCLUSION: The monotherapy with nifedipine GITS was effective in reducing high blood pressure, induced regression in ventricular hypertrophy and showed good tolerance in one year follow-up. PMID- 9340699 TI - [Decision making in cardiology: methodology]. AB - This article reviews the methods employed in usual clinical thinking for making decisions, the problems and limitations inherent in them and claims that a more frequent utilization of the so called "Evidence Based Medicine" methods is a more valid and efficient alternative for medical decision making. We also describe the theoretical basis and strategies used in the medical decision process; the specific concepts and basic components for building decision trees are also shown. Finally, a real case is presented and approached step by step: the statement of the decision problem, its possible alternatives, the allocation of probabilities to each outcome based on the best available evidence, and the calculations of the expected values (projected usefulness, cost-effectiveness) and sensitivity analysis by means of specific software for making decisions. PMID- 9340700 TI - [The syndrome of mid-ventricular obstruction with apical aneurysm in hypertrophic myocardiopathy: presentation of a case]. AB - We report an 81-year-old woman with hypertrophic cardiomyopathy, midventricular obstruction and associated apical aneurysm partially dyskinetic. At admission she showed a lateral acute myocardial infarction with sustained episodes of uniform ventricular tachycardia and subtle cardiac physical findings. Old apical infarction was suggested by resting thallium defects in the absence of obstructive coronary disease. The ECG revealed persistent ST elevation in the anteroapical leads without Q waves at discharge. This case report represents a rare example, in a previously asymptomatic elderly woman, of a distinct syndrome within the wide clinical spectrum of hypertrophic cardiomyopathy. PMID- 9340701 TI - [Extrinsic compression of the left atrium as an infrequent and fatal form of type B aortic dissection: value of ECG]. AB - We present an infrequent case of extrinsic compression of the left atrium caused by a type B aortic dissection diagnosed by echocardiography. The transthoracic echocardiography showed the obliteration of the left atrium by a heterogeneous mass with two circular images of low echogenicity. The TEE study identified the mass as a huge hematoma surrounding the true and false lumen of the aneurysm. The case demonstrated the value of echocardiography in the diagnosis of type B aortic dissection. PMID- 9340702 TI - [Ventricular tachycardia in a patient with mid-ventricular hypertrophic myocardiopathy and apical aneurysm]. AB - A case is presented of a 77-year-old patient who was admitted with a pattern of sustained ventricular tachycardia and diagnosed with midventricular hypertrophic myocardiopathy with apycal aneurysm. Under treatment with amiodarone at low doses, the patient is asymptomatic with no recurrence of the arrhytmias at one year. The association of midventricular hypertrophic myocardiopathy with apycal aneurysm and of those with sustained ventricular tachycardia are reviewed in conjunction with their treatment. PMID- 9340703 TI - [Biventricular diverticulosis associated with rheumatic heart disease: apropos of a case]. AB - We report a case of a 58-year-old woman with rheumatic mitral stenosis scheduled for percutaneous valvuloplasty. Prior left and right ventricular angiograms showed multiple diverticula at left ventricular apical and diaphragmatic walls and right ventricular diaphragmatic wall. Chest x-ray and echocardiogram were normal. Magnetic resonance imaging was concordant with catheterization findings and ruled out other cardiac malformations. The risk of ventricular perforation changed our indication of percutaneous valvuloplasty in favor of open heart commissurotomy. PMID- 9340704 TI - [Cardiac tamponade as a clinical symptom of systemic lupus erythematosus]. AB - Pericarditis is the most frequent cardiac manifestation of systemic lupus erythematosus (SLE), but pericardial effusion causing tamponade rarely occurs, and it is even less frequent for the pericardial tamponade to be the presenting feature of SLE. We report a case of pericardial tamponade due to SLE with severe hemodynamic involvement as the clinical presentation of the disease. We comment on its clinical course and its rarity in the medical literature. PMID- 9340705 TI - [Innovations in ambulatory nephrology]. PMID- 9340706 TI - [Pathogenesis of sodium and water retention in cirrhosis of the liver]. AB - Sodium retention is the most common renal abnormality of cirrhosis and eventually leads to the formation of ascites. There is now a strong body of evidence that the arterial vasodilatation, mainly splanchnic, that occurs during liver cirrhosis is a major factor in the pathogenesis of renal sodium and water retention. The arterial vasodilatation and the subsequent hypotension stimulate a baroreceptor-mediated neurohormonal vasoconstrictor and antinatriuretic response in a attempt to compensate the relative underfilling of the circulation. The cause of the arterial vasodilatation is thought to be a circulating factor and several recent studies have implicated an excessive vascular production of nitric oxide (NO). In animal models of cirrhosis administration of NO antagonists corrects the hyperdynamic circulation and improves renal sodium excretion. The treatment of sodium retention is often difficult in liver cirrhosis and it is possible that the identification of a vasodilator substance which can be inhibited could provide a new tool in the therapeutic approach of sodium and water retention in cirrhosis. PMID- 9340707 TI - [Differentiation between glomerular and non-glomerular erythrocyturia: what is the value of differential microhematuria diagnosis?]. AB - A few years ago, a new and simple method has been proposed to help guiding the investigation of microhematuria. This method which consists in quantifying the percentage of deformed polymorphous erythrocytes in the urinary sediment using phase contrast microscopy allows to distinguish glomerular from non-glomerular erythrocytes. In this paper, we have reviewed the recent literature concerning this approach and have discussed the conclusions according to our own experience based on the analysis of 147 patients presenting with microhematuria. Our results demonstrate that this technique is still limited by the difficulty to obtain well defined cut-off values which effectively differentiate renal from urologic diseases. Thus, only extreme results showing either the total absence or the presence of a very high percentage of dismorphic erythrocytes appear to be helpful for the physician. Despite the introduction of this new method, many patients with microhematuria are insufficiently investigated. PMID- 9340708 TI - [Renal manifestations of viral diseases]. AB - Like bacterial diseases viral diseases may also be accompanied by functional renal disorders or abnormalities of the urinary sediment. Thus, to find a hematuria or an isolated proteinuria in the context of influenza or hepatitis is not rare at all. In certain cases viral affections may even be accompanied by a nephrotic syndrome. This article aims at the discussion of multiple renal disorders appearing in the context of hepatitis B and C and AIDS. PMID- 9340709 TI - [Disorders of eating behavior--early detection and treatment possibilities in general practice]. AB - According to recent epidemiological investigations, one-third of younger women perceive themselves to be overweight. A quarter show disturbed eating habits such as continuous dieting, compulsive calorie counting or ruminations about eating and body weight. The ratio of young women to men with eating disorders is 10:1. Eating disorders, like other psychosomatic disorders, have a multifactorial origin. Biological, psychological, interpersonal and sociocultural factors are of varying importance in the development of the disorder. Apart from somatic aspects, the psychological component is of particular importance not only for the assessment of severity, but also for prognosis. Because patients with anorectic features often refuse help and patients with bulimia often suffer from a sense of shame and guilt, it is the physician's chore to note symptoms relevant to the clinical picture and to outline possibilities for therapeutic intervention. Furthermore, in the treatment of eating disorders it is important to recognise personal limitations and to decide upon appropriate treatment measures in collaboration with the patient when possible. PMID- 9340710 TI - [Fever and dry cough in a construction worker from Portugal]. AB - A 33-year-old Portugese worker presented with a one-week history of nonproductive cough and fever. A presumptive diagnosis "viral infection of the respiratory tract" was made. However, because of persisting cough and fever further investigations were necessary, and finally Brucella melitensis was isolated in blood cultures. Three months before admission to the hospital the man was dressing the carcasses of a goat in Portugal and consumpted fresh goats milk cheese. Antibiotic therapy with Rifampicin and Trimethoprim/Sulfamethoxazol over 6 weeks improved the signs and symptoms of the infection. PMID- 9340711 TI - [Chronic cough without expectoration]. PMID- 9340712 TI - [Cervical cancer screening]. PMID- 9340713 TI - [Cervix cancer screening: results of a french team]. PMID- 9340714 TI - [Cervical cancer screening. False negative smears]. PMID- 9340715 TI - [Predictive oncology and familial breast cancer]. PMID- 9340716 TI - [A look at neurology]. PMID- 9340717 TI - [Therapeutic networks]. PMID- 9340718 TI - [Smoking and chronic obstructive lung disease. What will stop respiratory function deterioration?]. PMID- 9340720 TI - [Prevention]. PMID- 9340719 TI - [Cardiology and heart surgery in the dawn of the 21st century. The viewpoint of policy makers]. PMID- 9340721 TI - [XXXVth French Congress of Oral Medicine and Maxillofacial Surgery. Montpellier, 17-19 September 1997]. PMID- 9340722 TI - [The sequellae of donor sites. Functional results of the treatment of cancer of the oral cavity]. PMID- 9340723 TI - [Spinal accessory nerve and lymphatic neck dissection]. AB - Radical neck dissection was the golden standard of treatment for cervical nodes in head and neck tumors. From the seventies, the preservation of the spinal accessory nerve has become increasingly popular in order to improve the functional result of the neck dissections. The aim of this study was to assess the degree of functional disability associated with each type of neck dissection and the value of anatomical references for dissection of the spinal accessory nerve. One hundred twenty seven patients were evaluated 1 month and 1 year after radical, functional or supraomohyoid neck dissection with a questionnaire and a physical examination. Anatomical measurements of the spinal accessory nerve were performed in 20 patients. We found considerable or severe shoulder dysfunction in 7%, 34% and 51% respectively of patients in whom supraomohyoid, functional and radical neck dissections were performed. Furthermore 49% of patients having undergone a radical neck dissection had little or no symptoms. Sacrifice of the spinal accessory nerve in radical neck dissection may lead to shoulder dysfunction. A functional disability may also be associated, although in a less extent, with any neck dissection in which the spinal accessory nerve is dissected and placed in traction. There is a large variation in the degree of functional disability and pain in patients with similar neck dissections. The course of the spinal accessory nerve in the neck makes it particularly vulnerable to injury during the dissection near the sternocleidomastoid muscle and in the posterior cervical triangle. PMID- 9340724 TI - [Management of cancer of the oral cavity: advancements and limits]. PMID- 9340725 TI - [Current status and prospectives in implantology]. PMID- 9340726 TI - [Post-traumatic temporomandibular pain dysfunction syndrome and the problems raised by its imputation]. AB - Direct, certain and total imputability of trauma as the cause of mandibular joint dysfunction is often difficult to establish. The theoretically obligatory conditions are rarely met, making the expert's opinion quite difficult to establish. The complexity of the pathophysiology involved in mandibular joint dysfunction confirms however the reality of this sequella. By determining the precise lesional or functional origin of the syndrome and the latent or patent nature of the prior status it is possible to ascertain intermediary situations. PMID- 9340727 TI - [Facial reconstruction: the history of a challenge]. AB - In our society, there is a social, cultural and legal obligation to identify cadavers. If all other techniques fail to produce a presumed identity for a very deteriorated body, facial reconstruction can be the last resort. Historically, the first attempts in the XIXth century concerned famous men. Anatomists, anthropologists, and embryologists established the basic principles of the method. Paleontologists then tried to reconstruct the face of prehistoric men. For the first time in the XXth century, the Russian school, directly inspired by the American school began work concerning the victims of crimes. The development of photography, the discovery of X-rays and progress in imaging and data processing, then the development of the CT scan have all contributed to this still experimental method. PMID- 9340728 TI - [Benefits and risk of the extraction of wisdom teeth]. PMID- 9340729 TI - [Experimental research in maxillofacial surgery]. PMID- 9340730 TI - [The facial approach to the median structures of the skull base]. AB - Numerous routes of access to the medial basal structures of the cranium have been described, largely because of the wide variety of lesions observed in deep localizations. Access can be achieve via trans-sinusal, transfacial (trans sphenoidal rhinoseptal, mediofacial or Lefort I), trans-oro-pharyngeal and numerous other routes. An examination of the principals involved, their development and the technical modalities demonstrate the advantages and disadvantages of each and their specific indications. Access is particularly interesting with the frontal trans-sphenoid, Lefort I osteotomy and trans-oro pharyngeal routes. The simplicity of these non-mutilating routes provide an alternative to neurosurgical access. Their development depends on progress in imaging and microscopic surgery. Used alone or in combination, they can be an useful complement to a neurosurgical access. PMID- 9340731 TI - [Periodontal surgery and guided tissue regeneration. Advancements in outcome:- concepts--means--applications]. PMID- 9340732 TI - [The prevention of infection in oral medicine]. PMID- 9340733 TI - [Tumors of the parotid gland]. AB - Tumors of the parotid are the most frequently encountered salivary gland tumors. Knowledge of the histology and anatomy of the salivary gland is important when considering the histiogenesis of salivary gland tumors, requiring close cooperation between the pathologist and the surgeon. Most tumors are benign epithelial formations. Pleomorphous adenomas predominate. Superficial lobectomy is adequate treatment. When the tumor involves a deep lobe, total parotidectomy is indicated. Treatment of malignant tumors depends on the histology, its TNM stage and other factors. Total parotidectomy with lymphadectomy and radiotherapy are needed in case of high grade malignancy. In children, vascular neoplasias are the most frequent, followed by malignant tumors. Their histological features and treatment are the same as for adults. PMID- 9340734 TI - [Ultrasound contrast media for neurovascular applications]. AB - Ultrasound is widely used in the assessment of neurovascular diseases. In spite of its effectiveness there are considerable limitations such as low flow detection in carotid disease or limited bony windows in transcranial Doppler. One approach to overcome these limitations is the use of ultrasound contrast enhancing agents. The usefulness of ultrasound contrast enhancing agents Levovist, EchoGen and BY 963 in neurovascular applications has been evaluated. Contrast enhanced colourflow Doppler for the diagnosis of carotid disease has been investigated in three small trials and might be effective for improving the diagnostic yield in severe disease. Contrast enhanced transcranial colourflow Doppler has been relatively more widely explored also with promising results. Based on the combined findings out of these preliminary investigational trials, it appears to be reasonable to undertake larger trials for assessment of usefulness of ultrasound contrast agents for a variety of neurovascular applications. PMID- 9340735 TI - [Possible errors caused by artefacts in the Doppler color image]. AB - The practical application of colour-coded duplex sonography shows that physico technical artifacts of colour Doppler imaging may not be obvious in all cases and can lead to misinterpretation. Slice-thickness artifacts are due to restricted transversal resolution, as in the B-mode, and may cause false negative results in vascular occlusions. Specific problems in creating and processing the colour flow image interfere with lateral resolution and lead to errors in the quantification of stenoses from the colour Doppler image. Mirror image artifacts produce phantom pictures that can be easily identified as such in some cases, but may resemble recesses in the arterial wall in other cases. Phenomena of sound shadowing resulting from the same reasons as in B-mode may lead to assume vascular occlusions in cases of long-range stenoses. Artifacts of the sonication angle in case of blood vessel tortuosity may lead to misdiagnosis of a non-existent retrograde flow. The artifacts mentioned do restrict the capacity of colour coded duplex sonography for localising flow in the ultrasound image. Users of the method should therefore be familiar with these artifacts. PMID- 9340736 TI - [A new method for imaging intracranial cerebral arteries--3-dimensional transcranial duplex ultrasonography with image enhancement]. AB - AIM: A new data acquisition system for transcranial 3 D-ultrasonography was used. The present study examines the evaluation of intracranial cerebral arteries by 3 D-ultrasound and echo contrast agents in comparison to cerebral angiography. METHODS: Ten patients, who underwent diagnostic cerebral angiography, were examined without knowledge of the angiographic results. Data acquisition through the transtemporal acoustic bone window was performed with a magnetic sensor system to track the spatial orientation of the ultrasound probe while scanning the volume of interest. We used a transcranial duplex system with power mode and stored a 3 D-data set before injection of transpulmonary stable contrast medium and afterwards. RESULTS: Ipsilateral to the transducer anterior cerebral artery (ACA) in 90%, middle cerebral artery (MCA) in 90%, three or more branches of MCA in 60% and posterior cerebral artery (PCA) in 40% were analysed without injecting an echo contrast agent. After the contrast injection ACA, MCA, branches of MCA and PCA in 100% were visible. In 70% of cases the posterior and anterior communicating arteries were visualised only by means of a contrast agent. CONCLUSION: Application of a transpulmonary stable ultrasound contrast agent in combination with 3 D transcranial duplex ultrasonography significantly enhances the visualisation of intracranial arteries. PMID- 9340737 TI - [Value of Doppler color ultrasonography in diagnosis of arterial occlusive disease of the lower extremity. A direct comparison with percutaneous angiography]. AB - A Direct Comparison with Percutaneous Angiography: AIM: To compare colour-coded Doppler sonography (CCDS) with conventional angiography in severe occlusive vascular disease of the lower limb. METHODS: In 55 patients 1141 vessel segments were evaluated, 700 of them with atheromatous plaques, 270 with stenoses, 208 with occlusions and 6 with aneurysms. RESULTS: Deeper-seated vessels such as the abdominal aorta, the pelvic arteries, the superficial femoral artery at the level of the adductor canal and parts of the lower leg arteries are less accessible for direct CCDS. Many pathological changes however can be diagnosed indirectly by changes in the spectral wave form distal to the lesion. In superficial vascular segments (the common femoral artery, the profunda femoris artery, the superficial femoral artery above the adductor canal and the popliteal artery) image quality was excellent, more pathological changes were found, and the degree of stenosis was better estimated in comparison to angiography. CONCLUSION: The value of CCDS in patients with intermittent claudication is limited to those who have been examined with angiography e.g. before angioplasty, to follow-up examinations after vascular dilatation or surgery and to supplementary visualisation after angiography especially in readily accessible (superficial) vascular segments. PMID- 9340738 TI - [Ultrasonography of Achilles tendon lesions--an experimental study]. AB - AIM: In an experimental study the accuracy of ultrasound in the detection of lesions of the achilles tendon was examined. METHODS: 60 Achilles tendons were examined post mortem macroscopically, histologically and by means of ultrascan. RESULTS: All macroscopical changes were found in the sonograms. Especially changes of the form proved to be good diagnostic patterns for degenerative lesions of the tendon. At a distance of 2 cm proximal to the calcaneus histological changes could be detected with an accuracy of 73.3%. At 4 and 6 cm to the calcaneus we found similar results. CONCLUSION: A differentiation of the histological diagnosis by means of the sonographic behaviour was not possible. As a reproducible, effective and inexpensive method ultrasound examination has proved very useful in the detection of degenerative lesions of the Achilles tendon. PMID- 9340739 TI - [Ultrasound findings after shoulder dislocation]. AB - AIM: Sonography of the shoulder is an effective method for detecting tears of the rotator cuff and bone lesions. The purpose of this study was to evaluate prospectively the sonographic findings after shoulder dislocation. METHODS: Sonography was performed on 208 patients with 210 shoulder dislocations, which were all verified by radiography. RESULTS: We diagnosed 62 rotator cuff lesions (29.5%), which were all in the group with traumatic shoulder dislocation. The incidence of tears in patients with first traumatic dislocation (n = 134) was 36.5%. 12 tears (21.8%) of the rotator cuff were noted in recurrent dislocations (n = 55). 23 fractures or bony avulsions of the greater tuberosity of the humerus (11%) and 168 Hill-Sachs lesions (80%) were seen. CONCLUSION: Sonographic examination of the shoulder revealed a significant amount of information that would remain undetected without the aid of expensive and/or invasive diagnostic tools. PMID- 9340740 TI - [Transabdominal chorionic villi and placental biopsy: rapid karyotyping in the 1st-3rd trimester of pregnancy]. AB - AIM: The diagnostic value and the complication rate of transabdominal chorionic villi and placental sampling was compared with standard amniocenteses. The method ist especially helpful in cases with conspicuous fetal sonographic findings. METHODS: The results of 475 biopsies were retrospectively compared with 983 amniocenteses and chorionic villi samplings (CVS). RESULTS: 64% of chorionic villi samplings (CVS) were performed in the first, 30% in the second and 6% in the third trimester. The indications were advanced maternal age (45%) and psychological problems (14%) in the first trimester and conspicuous maternal serum markers (11%) or fetal ultrasound anomalies (12%) in the second and third trimester, respectively. 10 out of 20 aneuploid cytogenetic results were found in fetuses with sonographic anomalies. In 4 cases we found confined placental mosaicism, which was clarified by means of amniocentesis and cordocentesis. We had 8 miscarriages in a total of 475 CVS procedures; 6 in 304 before the 15th week of gestation (1.97%). The natural abortion rate in this gestational age is about 1%, the CVS-related abortion rate therefore is near 1%. CONCLUSION: Transabdominal CVS is a low risk method for rapid karyotyping during the entire pregnancy. PMID- 9340741 TI - [Ultrasonography of the normal vermiform appendix]. AB - AIM: To study the detectability and the appearance of the vermiform appendix with ultrasound in asymptomatic adults. METHODS: A prospective study was performed on 300 patients, selected without regard to age, sex and weight. We tried to visualize the appendix with a 3.5 MHz annular array and a 5 MHz linear array transducer. RESULTS: In 63% of the patients the appendix was clearly visualised. The main reasons for non-visualisation were obesity with insufficient ultrasound penetration (38%) and the caecum laying in pelvic or atypical position (24%). In 31%, no cause was apparent but we assume that the appendix often lies behind the caecum. The sonographic image showed an aperistaltic target originating at the caecum, and ending blindly. In 42% of cases the appendix was ovoid in transverse section; in 32%, round; and in 26% both forms were found within one appendix. Intraluminal gas was visible in 86%. The mean transverse diameter was 5.2 mm (min. 3 mm, max. 13 mm) and in 76% the transverse diameter was lower than 6 mm. CONCLUSION: The sonographic detectability, the appearance and the size of the normal appendix show considerable variations. PMID- 9340742 TI - [Recent developments in the area of Doppler ultrasound imaging]. AB - Developments in Doppler Sonography: Review of duplex Sonography with and without ultrasonic contrast media, power Doppler, transcranial Doppler sonography. Duplex sonography for investigating the extracranial brain circulation should always be used in conjunction with unidimensional direction sensitive Doppler sonography. PMID- 9340744 TI - Work in clinical research--without a medical degree. PMID- 9340743 TI - The science of substance abuse. PMID- 9340745 TI - Polio vaccine production. PMID- 9340746 TI - Clean air skepticism. PMID- 9340747 TI - Genetic evolution of morphology. PMID- 9340748 TI - Resonance Raman results. PMID- 9340749 TI - Weakened SIV vaccine still kills. PMID- 9340750 TI - Furor over company deal roils AMA. PMID- 9340751 TI - Stanford researcher fights dismissal in hospital spat. PMID- 9340752 TI - Government bows out of Academy case. PMID- 9340753 TI - New studies affirm BSE-human link. PMID- 9340754 TI - Near the lights of Vegas, chemists are on a roll. PMID- 9340755 TI - Males mutate more, bird study shows. PMID- 9340756 TI - Getting the brain's attention. PMID- 9340757 TI - Cocaine wreaks subtle damage on developing brains. PMID- 9340758 TI - MutT prevents leakiness. PMID- 9340759 TI - Unconscious odors. PMID- 9340760 TI - Proteins from scratch. PMID- 9340761 TI - Tobacco control: the dramatic choice. PMID- 9340762 TI - Man and his dog. PMID- 9340763 TI - Man and his dog. PMID- 9340764 TI - Flap on NEJM board over ethics articles. PMID- 9340765 TI - Need a reagent? Just sign here... PMID- 9340766 TI - Prusiner recognized for once-heretical prion theory. PMID- 9340767 TI - AIDS research chief bows out. PMID- 9340768 TI - How the malaria parasite manipulates its hosts. PMID- 9340769 TI - Fishing for answers to whirling disease. PMID- 9340770 TI - Io and the plasma torus. PMID- 9340771 TI - Reuse of B lymphocytes in germinal centers. PMID- 9340772 TI - Good pain, bad pain. PMID- 9340773 TI - BMP expression in duck interdigital webbing: a reanalysis. PMID- 9340774 TI - [Rational surgical management improves outcome in tibial plateau fractures]. PMID- 9340775 TI - [Tibial plateau fractures]. AB - The classification of tibial plateau fractures is based on morphological criteria according to AO/ASIF and Schatzker or on functional criteria according to Moore. A total of 81 acute tibial plateau fractures were operated on over a 6-year period. The patient data and the operative procedure are given (not including late results). Some 6% of ligamentous lesions were treated conservatively. Bicondylar osteosynthesis was performed in 11%, and 14% of the operations were minimally invasive procedures. In 61.7% the plateau was fixed by a buttress plate. Autologous cancellous bone from the iliac crest was implanted into the fracture, especially into depressed wedge fractures (Type AO/ASIF B3 or Moore Type IV). In 39.5% of the cases a corticocancellous wedge-shaped bone chip from the iliac crest was used. Complications were rare, and no infection was observed. Most important are the soft tissue balance, the timing of surgery, choice of longitudinal incisions directly above the lesion, and a stabilization procedure which maintains blood flow and coverage of the fracture. The operative procedures are described including tibial plateau fractures in children and the management of complications. In special situations a conservative treatment is still useful. PMID- 9340777 TI - [Arterial vascular lesions after total hip joint replacement]. AB - We report on three arterial thromboses of the external iliac artery following total hip replacement. As a result of implanted cement/spongiosa or protrusion of the acetabular component, the iliac vessels were compressed. Furthermore, we report about one intraoperative arterial vessel lesion in a 65-year-old patient during a revision operation. We recommend that in case of acute ischemic syndromes of lower limbs following total hip replacement, an angiography should be performed in order to exclude an extravascular cause of thrombosis. For therapy in those cases extra-anatomic bypasses should be preferred to thrombectomies. PMID- 9340776 TI - [Magnetic resonance tomography examination of thoracolumbar spinal fractures after fixateur interne stabilization]. AB - To analyse the possible injuries of vertebral segments, especially the disc, after unstable thoracolumbar fractures stabilised with AO internal fixator, we performed magnetic resonance imaging (MRI) of the traumatised region after implant removal. There were two aspects of disc degeneration (DD):(1) biochemical changes and (2) structural damage. MRI detects biochemical processes as one aspect of DD that is often small even in the presence of greater structural damage of the nucleus pulposus caused by fracture. None of the patients presented with structural failure of the anulus fibrosus, which is the essential structural component of the vertebral segments with regard to stability. We observed biochemical changes more often in the lower of the two fracture-adjacent discs and alterations of discal shape more often in the upper of the two, whereas loss of height concerned both discs to approximately the same degree. The supporters of upper-disc resection in thoracolumbar fractures justify their procedure among other things with the structural disc damage, such as alteration of shape and loss of height (altogether more frequent in the upper disc). Our observations that a disc with a structurally altered nucleus pulposus can be biochemically intact and can show an intact anulus fibrosus are arguments in favour of disc preservation. With regard to the upper disc, the widespread opinion that complete and regular disc damage requires a resection has to be revised. The question of whether the lower disc should be resected more often because of its greater biochemical changes cannot be answered by the present study alone. Besides the excellent static information in all anatomical structures of the vertebral column available by MRI, a repeat examination in a prone position yields dynamic information on the spinal cord in the case of suspected dorsal adhesions. PMID- 9340778 TI - [Percutaneous therapy of osteoid osteoma]. AB - Osteoid osteomas are tumors with intense clinical symptoms and extensive reactive bone changes far exceeding the volume of the lesion itself. Because of their small size they can be approached by minimally invasive surgical procedures. We treated ten symptomatic patients with osteoid osteomas (n 6 hip point, n 1 iliac bone, n 1 femoral diaphysis, n 2 tibial diaphysis) by excision of the nidus with a 3-mm Harlow-Wood needle using a percutaneous CT-guided approach. Seven patients with residual tumor were treated with either thermocautery (n 2) or sclerosis with 1 ml of 96% ethanol (n 5). Six patients had instant and constant relief (3 years' observation) of their pain. In two patients a second transcutaneous intervention was successful. Only two patients needed open resection. Compared with the invasive open resection of the tumors, sometimes even putting the stability of the femoral neck at risk, transcutaneous CT-guided enucleation of the nidus of the osteoid osteoma with additional sclerotherapy is a good alternative method, especially in the region of the femoral neck. PMID- 9340779 TI - [Effect of surgical technique on meniscus transplants. A histological, animal experiment study]. AB - After transplantation, a meniscus undergoes alterations in mechanical loading, which causes changes in its histological structure. We studied the degenerative effects on meniscus and tibial cartilage resulting from variations in the congruity or the isometric fixation of medial meniscus transplants. In three groups of five sheep each, the menisci were transplanted in three different ways, using the same operative approach. The menisci were evaluated 24 weeks after operation. In group 1, the meniscus was totally detached from its base at the capsule and refixed without changes in the congruity or isometry. This group provided the basic data. In group 2, the contralateral medial meniscus was turned upside down and transplanted. The reattachment was performed according to isometric conditions. With this technique the congruity of the tibial and femoral surface was modified. In group 3, the medial meniscus was reimplanted by choosing defined non-isometric fixation points for the anterior and posterior meniscal ligaments without changing the position of the corpus. For evaluation, the morphological alterations of meniscus and tibial cartilage were assessed by the Jackson score. The more distinct changes of the meniscus were assessed histologically by three criteria: surface cells, surface fibers and changes in the meniscus center. The highest degree of degenerative changes occurred in group 3 (score 4.5); however, considerable changes were also found in group 2 (score 3.5). Incongruous or non-isometric placement of a meniscal graft will lead to degeneration and failure of the graft. PMID- 9340780 TI - [Process and outcome quality in giant cell tumor in relation to surgical management]. AB - Giant-cell bone tumors display a locally aggressive growth pattern, frequently recur if no adjuvant treatment is given, and may potentially metastasize. By virtue of their biological behavior and typically juxta-articular localization, giant-cell bone tumors require specific surgical management. Thus, an intralesional tumor excision must be supplemented by adjuvant bone cementing, possibly combined with instillation of phenol or cryotherapy. These combined treatment modalities assure a high-quality procedure, defined as the actual way medical care is delivered, by promoting the quality of the outcome, defined as the effect of a medical procedure on the patient's state of health. PMID- 9340781 TI - [Functional, neuropsychological and social outcome of polytrauma patients with severe craniocerebral trauma]. AB - The aim of this study was to identify, in (pre-) clinically obtained data, parameters predicting the outcome of patients with multiple trauma and severe head injury. Fifty-eight patients aged 27 +/- 10 years were investigated an average of 5.8 years after the accident. The Hanover Polytrauma Score was 34 +/- 11 points, the initially assessed Glasgow Coma Scale (GCS) was 6.2 +/- 3.2 points; and the duration of coma was 15.4 +/- 14.4 days. The primary length of stay in hospital averaged 33.4 days, including 22.9 days in the intensive care unit and 20.2 days of ventilation. For a further 223 days the patients were treated at the Neurologic Clinic of Hessisch Oldendorf. Besides different neurologic deficiency symptoms, the psychometric tests showed deficits in all areas. In particular, information processing speed, concentration, recent memory and learning performance were impaired. There was free mobility of all joints in 33% of the patients. Due to injury the elbow and ankle joint developed the worst restriction. Central paralysis and heterotopic ossification also caused a restriction in joint mobility. Half of the patients were confronted with different social changes. The rate of return to work was dependent on age. Some 42% of all patients had taken up their former profession, 5% were still in training or at college, 32% were retrained to other professions, 16% were unemployed and 5% were completely retired on pension. Age, injury severity, GCS, duration of coma and duration of weaning were suitable predictors in correlation- and regression analysis. The Glasgow Outcome Scale showed good recovery and moderate disability in 53%, severe disability in 33% and persistent vegetative state in 14% of the patients. PMID- 9340782 TI - [Automobile head supports--adjustment possibilities and utilization. Results of a field study]. AB - An important detail referring to the whiplash syndrome is the relationship between the position of the head restraint and the head, because the head restraint protects the head in case of a rear-end accident. This relationship was evaluated in a representative study in 601 nonselected volunteers. A horizontal distance between the head and head restraint of maximal 9 cm was present in 69.6%. An optimal distance of 0 cm was only found in 7.4%. Just 50% of the adjustment distance was used by the car drivers. Even in completely distracted adjustments in 50% a deficit of more than 8 cm, and in 20.9% a deficit of even more than 12 cm was present. These results show that passive protection, on the one hand, is not guaranteed because of the lack of proper height adjustment. On the other, the volunteers did not use the best adjustment each time. PMID- 9340783 TI - [1996 Annual Meeting of the Orthopedic Trauma Association]. PMID- 9340784 TI - [Injuries to the growth plates]. PMID- 9340785 TI - [Saving the extremity by massive transfusion in grade III open femoral fracture with vascular lesion. A case report]. AB - The major problems in the treatment of open fractures with high blood loss are hypothermia acidosis and coagulopathy. By improving the standards of polytrauma management and using massive transfusion systems, which should help to avoid those complications, the indication for primary limb salvage under hypovolemic shock conditions is greatly increased. The following case of an grade 3 open fracture of the femur with severe soft-tissue damage, including vascular lesions, with consecutive massive transfusion, should demonstrate that the cardiac respiratory system can still be stabilized, even if a surgical solution seems impossible. PMID- 9340786 TI - [Successful surgical revascularization after 18 hour ischemia below the knee due to complete avulsion of the popliteal artery]. AB - We report the case of a 33-year-old man after knee luxation and disruption of a popliteal artery, which was missed after repositioning at the first clinic. Eighteen hours later complete discontinuity of the popliteal artery was confirmed by arteriography and an immediate reversed, end-to-end auto-venous graft was interposed under general anesthesia with anterior and posterior fasciotomy prior to blood flow restoration. Forced diuresis with a diuretic mixture and balanced fluid intake were used, and the patient was discharged from the intensive care unit on the 10th postoperative day in good condition with normal diuresis. PMID- 9340787 TI - [Value of diagnostic laparoscopy in abdominal trauma]. PMID- 9340788 TI - The Fox Chase Cancer Center and Free University Hospital Investigators' Workshop and Consensus Conference on Paclitaxel. Part 1: Lung Cancer. Puerto Rico, March 12-16, 1997. PMID- 9340789 TI - [A case of fatal poisoning with the anti-arrhythmia agents Katen and Neo Gilurytmal]. AB - The case is presented on 17-year-old student who ingested larger doses of tablets and capsules in privacy. She ingested antiarrhythmics Katen and Neo-Gilurytmal without clinical therapy by these drugs and without history of suicidal attempt. The imminent case of death was asphyxiation by aspiration of vomits from gastric contents after ingestion of excessive doses of drugs. The secondary findings as brain edema, petechiae under the serous membranes and congestion of the abdominal cavity, were relieved at autopsy. Noxae was identified by the postmortem toxicological analysis of blood, liver, kidney, gastric contents and intestinal contents. Mexiletine and prajmaline were analysed by the capillary gas chromatography with FID detection. Retention time of mexiletine was 6.66 minutes, prajmaline 15.15 minutes, respectively. Blood alcohol concentration was 0.21 mg.g 1. Concentration of prajmaline in gastric contents was 3.03 mg.g-1, mexiletine 0.11 mg.g-1, respectively. PMID- 9340790 TI - [Detection of drugs using water in high performance thin-layer chromatography]. PMID- 9340791 TI - [Immunohistochemical detection of myocardial hypoxia]. AB - Antibodies against myoglobin and fibrinogen were used for immunohistochemical investigation of myocardium in a group of 45 persons deceased of natural and violent cause. Suffocated persons showed a decrease or disappearing of myoglobin in 66% and disseminated positivity for fibrinogen in myocardial fibres in 70%. Deceased of violent cause without hypoxia had disseminated decrease or disappearing of myoglobin and simultaneous disseminated positive reaction for fibrinogen but in 14% of cases. Deceased persons with macroscopical heart infarction were used for control and had focal lack of myoglobin in 100%. Immunohistochemical investigation of myoglobin and fibrinogen can be used as a very sensitive prove of prolongated hypoxia or very intensive asphyxia when positive signs can occur after a very short period. PMID- 9340792 TI - [Questions from a medico-legal viewpoint on the planned law dealing with removal and transfer of organs (transplantation law) in West Germany]. AB - Within the framework of the planned law concerning the removal and transfer of organs (transplantation law) in the Federal Republic of Germany, and taking as a basis the draft of a prototype law by the federal states dated February 1st 1994 and the draft bill of the states of Bremen and Hessen dated June 30th 1994, the speech addresses itself primarily to the question of whether, when no consent of the decreased can be identified, and when the next-of-kin have been informed by the doctor in attendance and given an appropriate time-limit, the next-of-kin in order of succession or any close relative can object to organ removal for the purpose of transplant surgery. Because of the highly personal nature of the postmortal personal rights transferred to the next-of-kin as well as their custodial rights, and with regard to the particularly sensitive nature of this decision, it is suggested that the next-of-kin be informed not only of their right of refusal but also-in view of their responsibility toward other relatives be instructed on the meaning, substance and legal consequences of such a refusal. It is furthermore suggested that sufficient organs could be obtained by organisational measures such as requests to driving licence applicants, which would lead to a higher preparedness to donate organs, while, at the same time, sparing the next-of-kin unnecessary emotional pain. From a medical point of view, it should be recognised that the complete and irreversible loss of all brain functions, in spite of artificially maintained heart and circulatory functions, is a sure sign of death. In closing, it is pointed out that a change to the currently valid [symbol: see text] 168 StGB would mean that a violation of the right of refusal could lead to criminal prosecution. PMID- 9340793 TI - [Voice problems in some groups of professional users of voice: implications for prevention]. AB - The primary prevention of communication problems is a growing part of the professional responsibility of the speech-language therapist. Functional voice hyperfunction, which often occurs in professional users of voice, can be prevented. In this article three separate studies on voice problems of people in three different professions are reported. The subjects included 183 educators, 50 ministers and 20 singers. The data were obtained using questionnaires. The results indicate that voice problems and symptoms of voice problems occur in all three groups, that these speakers have little knowledge of the subject, that they expose themselves to high risk factors, and that they do not conserve their voices. Guidelines for instruction in prevention are provided. PMID- 9340794 TI - [Spoken and written language abilities of remedial class children placed back into mainstream education]. AB - Current literature emphasizes the role of the speech-language therapist with regard to language learning disabled children. A lack of research regarding the language abilities of the older language learning disabled child was, however, identified. The possibility of continued spoken and written language problems in aid class children who have been placed back into mainstream education, served as a motivation for this research project. The empirical survey was administered on twenty-eight Afrikaans and English speaking ex-aid class children. According to experimental aims the subjects' level of performance in spoken and written language and scholastic skills were analysed and described qualitatively and quantitatively, through the Test for Oral Language Production and the South African Written Language Test. The findings indicate that a significant percentage of the subjects evidenced spoken and written language problems and moderate difficulties in the school subjects Afrikaans and English. Meaningful correlations were found to exist between spoken and written language, and language skills and scholastic performance. Implications for dealing with the language learning disabled child are discussed, and continuation of speech language intervention with aid class children who have been placed back into mainstream education, is strongly recommended. A need for further research with regard to the older language learning disabled child is expressed in order to support and expand these results. PMID- 9340795 TI - [Changes in intraocular pressure during dialysis]. AB - We studied 29 patients on dialysis for terminal uremia, of whom one showed a dramatic rise in intraocular pressure (IOP) during dialysis. We examined the blood pressure (BP) and body weight (BW) before and after treatment in each patient and visual acuity by gonioscopic examination and optic disk status. After dialysis the statistically significant fall in BP and BW was observed in all patients, and in 28 subject an significant rise in IOP was noted. Only one patient experienced a dramatic rise in IOP, and he was the only one suffering from angular glaucoma during the last two years. All this time he was continually treated for glaucoma. After dialysis his IOP was 45 mmHg. With the adequate medication, his IOP lowered to the point when he was able to bear iridectomy which was done with argon and YAG laser. After the surgery his IOP reached the normal values. His visual acuity was good (1.0), visual field was normal, gonioscopic findings showed a narrow angle (Shaffer I, with normal pigmentation), and the optic disk had no abnormalities. It is not yet clear what was the cause of IOP rise during dialysis. It might be that the increased production of aqueous humor, which has been reported in such cases, might be caused by imbalance between blood and aqueous osmolality. If dialysis causes decrease in blood osmolality and if, at some time, blood becomes hypotonic in relation to ocular fluid, then the regular osmotic forces transfer the fluid to the ocular fluid into the eye. This case suggests that all patients on dialysis should undergo a minute ocular anamnesis and examination, and if needed administered adequate medication during the treatment. PMID- 9340796 TI - [Acute superficial thrombophlebitis--modern diagnosis and therapy]. AB - Acute superficial thrombophlebitis of the lower extremities is one of the most common vascular diseases affecting the population. Although it is generally considered as a benign disease, it can be extended to the deep venous system and pulmonary embolism. We examined 50 patients (22 males and 28 females), mean age 52.5 years. These patients were surgically treated due to acute superficial thrombophlebitis of the lower limbs that affected great saphenous vein above the knee. The diagnosis was made by palpable subcutaneous cords in the course of great saphenous vein or its tributaries in association with tenderness, erythema and oedema. Of these 50 patients, 26 were examined by duplex ultrasonography before the operation. In 20 patients duplex scanning confirmed that the process was greater than we supposed after clinical examination (77%) and in 6 patients there were no differences (23%) (Figures 1 and 2). The operation included crossectomy, ligation and resection of the proximal part of the great saphenous vein. Intraoperative findings in 38 patients showed that the level of the phlebitic process was higher than the clinical level (76%). There was no difference in 12 patients (24%). Deep vein thrombosis and pulmonary embolism were noted in 14 patients (28%) (Tables 1 and 2). Both complications were found in two patients, and 12 had one of these complications. Generally, there were 12 patients with deep venous thrombosis and 4 patients with pulmonary embolism. Only in one patient deep venous thrombosis appeared postoperatively, while all other complications occurred before surgical intervention (Scheme 1 and Table 3). The most common risk factor was the presence of varicose veins (86%). Obesity, age over 60 years, cigarette smoking are listed in decreasing order of frequency. Patients under 60 years were more likely to have complications while older patients usually followed a benign clinical course (Tables 4 and 5). There was no intrahospital mortality. Average hospitalization was 5.7 days. It was 4 days in patients without complications. After thes urgent operation that practically removed the risk of potentially fatal consequences, the patients were dismissed from hospital. New hospitalization was recommended after two weeks when the second act of surgical treatment was performed. It included stripping of the great saphenous vein and extirpation of varicose veins in the area without acute inflammation. The findings of this study confirm the general opinion that acute superficial thrombophlebitis is a very common vascular disease with usually "benign" clinical course. In its ascending form that affects the great saphenous vein above the knee it can be associated with deep venous thrombosis and pulmonary embolism. The level of phlebitic process is usually much higher than can be palpated clinically. Duplex scanning was a highly reliable, precise, fast non-invasive diagnostic method that is necessary in examining, following and making decision for operative treatment of acute superficial thrombophlebitis. If suspected complications an urgent surgical intervention should be performed. It is short and efficient, contributing to the fast recovery of the patients and their return to normal activities. PMID- 9340797 TI - [The effect of cigarette smoking during pregnancy on fetal growth]. AB - The association between maternal smoking and poor pregnancy outcome, which is well established in medical literature, has been confirmed during the study conducted in one of Belgrade hospitals. The study comprised 1011 women who gave birth to a live born baby between June 1992 and March 1993 (infants with malformations were excluded). The women were interviewed by one person during the first three days after delivery. Data were collected on smoking habits during the first, second and third trimester of pregnancy, and on potential confounders including age, education, marital status, obstetric history, height, weight before pregnancy, weight gain during pregnancy, history about diseases before and during pregnancy, housekeeping and occupational activities, data about delivery and data about alcohol consumption. By the use of factor analysis infants characteristics, taken from medical histories, were classified in two groups: I group-birth weight, birth length, head circumference and chest circumference; II group-apgar score after one and after five minutes. The rough relation of smoking to the outcome was examined first. Adjustment was made for the potential confounders by the use of multiple regression analysis. Variables associated with both smoking and birth weight or apgar score after one minute (i.e. maternal height and weight) were considered as potential confounders. Infant sex, gestational age and parity were also included as possible confounders due to their strong link with birth weight and apgar. In the sample 42% of women were smokers, and 98% of them smoked filter cigarettes. In the group of smokers 312 smoked throughout pregnancy and 111 were inconsistent smokers (those who smoked during one or two trimesters only). According to smoking habits, mothers were divided into three groups: nonsmokers, those who smoked 1-9 cigarettes per day and those who smoked 10 or more cigarettes per day (Table 1). There was a strong relationship with significant linear trend between smoking and all observed birth outcomes except apgar score (Table 2). Significant reductions in birth weight (by 205 g), birth length (by 1.28 cm), head circumference (by 0.38 cm) and chest circumference (by 0.66 cm) were found to be associated with an average daily smoking of 10 or more cigarettes after adjustment was made for potential confounders. Even a smaller number of cigarettes affected foetal growth. Infants born by mothers who throughout pregnancy smoked 1-9 cigarettes per day (mean 4.07, range 1-8) weighted significantly less (by 89 g) and had head circumference significantly smaller (by 0.23 cm) in comparison with infants born by mothers nonsmokers. Inconsistent smoking during pregnancy had no significant effect on foetal growth with the exception of smaller birth length (by 2.30 cm; p = 0.002) in infants born by mothers who smoked during the second and third trimester. It is possible that the relatively small number of women inconsistent smokers had a bearing on the results. According to the results obtained it seems that either there is no threshold for the effect of smoking on foetal growth, or it is very low. Nevertheless, since the effect of smoking is weaker if the number of cigarettes smoked is smaller it is reasonable to assume that the reduction in the number of cigarettes smoked in pregnancy would serve as prevention irrespective whether the threshold existed or not. PMID- 9340798 TI - Prevalence of various levels of obesity in individuals on diet therapy. PMID- 9340800 TI - [Extracorporeal shock-wave lithotripsy in patients with manifestations of urinary tract infection]. AB - The urinary tract infection is very frequent, especially if calculosis of the urinary tract is present. Urinary infection is widespread, and it appears during the year. The people of all ages and both sexes are affected by urinary infection. In the last few years a reliable progress in the understanding and management of urinary tract infection is achieved. Numerous articles published in professional journals are a good proof of it. The urinary tract infection is frequent and is responsible for the use of large quantities of antibiotics which provoke great costs and make other problems. The role of laboratory tests in the diagnosis of infection is predominant. The clinician is completely dependent on his collegue, a bacteriologist, with regard to the results of urine culture. It is known that microorganisms grow better if they have good nourishment. Infections of the urinary tract were always a significant problem. However, over the last few decades, they became, according to some authors, the most frequent bacterial infection in humans, requiring the frequent administration of immunosuppressive agents, corticosteroids and cytostatics; and at the same time a great number of elder people and chronic patients with reduced immunity are involved. Taking into account that significant and insignificant infections of the urinary tract are frequent in nephropathology, particularly in renal and canalicular calculosis, the aim of the study was to point to extracorporeal shock wave lithotripsy without risk of impairment of already existing infections with and without administration of antibiotic and uroantiseptic agents for prophylactic purposes. A group of 5,078 patients with calculosis of the urinary tract was studied. Extracorporeal shock wave lithotripsy was performed in all patients by Siemens lithotriptor Lithostar (Germany). In patients with calculosis of the urinary tract subjected to extracorporeal lithotripsy bacteriuria was regularly followed. A group of 1,836 (36 percent) patients with urinary tract obstruction and 3,242 (64 percent) patients without urinary tract obstruction were treated (Table 1). In 895 (18 percent) patients with urinary tract obstruction infection was serious. In 321 (6 percent) patients without urinary tract infection, serious urinary tract infection was detected (Table 2). The most frequent causes of urinary tract infection are presented in Table 3. Table 4 shows a review of patients to whom antibiotic therapy, prior to extracorporeal lithotripsy, was prescribed. Infection of the urinary tract is responsible for great morbidity. The treatment of any type of urinary tract infection must include the examination of the effect of antibiotic agents. During the treatment of urinary tract infection with calculosis resistant microorganisms are also developed because of repeated administration of antibiotics to patients in health institutions, and especially to patients with ureteral catheters. The treatment of any type of urinary tract infection must include the examination of the effect of antibiotic agents used. The fundamental aims of the treatment of urinary tract infection are: the eradication of causes of infection and concurrent prevention or optimal control of recurrent infection. As long as the patients with urinary tract calculosis are susceptible of permanent infections. It is indispensable to perform sterilization, and thereafter to remove the stone from the urinary tract, because infection of the urinary tract may cause a series of sequelae in the function of the kidney. Frequently the successful urinary sterilization with antibiotic agents cannot be achieved, and consequently, the carrying out of extracorporeal lithotripsy together with administration of antibiotics, is impossible. Good results can be obtained by a combined therapy of antibiotics and extracorporeal lithotripsy in patients with urinary tract calculosis. (ABSTRACT TRUNCATED) PMID- 9340799 TI - [Carotid body tumor]. AB - INTRODUCTION: The carotid body tumour was first described by von Haller in 1743. The first two, unsuccessfully surgically treated carotid body tumours, were done by Reinger in 1880 (his patient died), and by Maydel in 1886 (his patient developed hemiplegia). Scudder made the first successful surgical removal of the carotid body tumour in 1903. Using data from the Cologne (Germany) Medline Research Centre, surgical treatment of carotid body tumour was not reported in Yugoslav medical literature. The aim of this study is to present 6 surgically treated carotid body tumours. MATERIAL AND METHODS: Over the period from 1982 to the end of 1996, 6 patients with carotid body tumours were operated on in the Centre of Vascular Surgery of the institute of Cardiovascular Diseases of the Clinical Centre of Serbia in Belgrade. Four of them were female and two male patients, average age 43.4 years. In all cases the tumour was an asymptomatic neck mass. Color-Duplex ultrasonography and selective carotid arteriography were used to establish the diagnosis in 5 cases. The pathohistological examination of all 6 patients revealed the benign character of tumors. Patient 1. A 52-year old man. The suspicion of symptomatic carotid artery aneurysm, was the indication for urgent operation. The intraoperative finding showed a carotid body tumour which compressed carotid arteries. The subadventitial removal of the tumour was done. The patient was followed for 14 years without signs of local recidivation. Patient 2. A 38-year old man. During the operation the tumour was removed subadventitially, without clamping or injuring the carotid arteries. The patient was followed for 8 years and 3 months, and there were no signs of local recidivation. Patient 3. A 48-year old woman. Intraoperative findings showed an infiltration of the carotid arteries and tumour was removed together with parts of internal and external carotid arteries. The internal carotid artery was reconstructed using saphenous vein graft. The follow-up period was 4 years and 6 months, without signs of local recidivation. Patient 4. A 61-year old woman was operated on (neck exploration) in other hospital 4 years before the admission to our Centre. During the primary operation, an internal carotid artery was ligated without neurological consequences. Also, histological examination was performed. We removed a tumour together with the ligated internal carotid artery without its reconstruction. Three years after the operation the patient was without signs of local recidivation. Patient 5. A 40-year old woman. After subadventitial surgical removal of the tumor without clamping or injuring the carotid arteries, the patient was followed-up for 2 years and 2 months, and was without signs of local recidivation. Patient 6. A 30-year old woman was operated on (neck exploration only) in other hospital two months before the admission to our Centre. Intraoperative findings showed tumour infiltration to the carotid arteries, and therefore, internal and external carotid arteries were removed together with the tumour. The internal carotid artery reconstruction was performed using aaphonous vein graft. The early postoperative period was unremarkable. However, 48 hours after the operation cerebrovascular insult developed with hemiplegia. There was no sign of graft thrombosis. The patient was followed-up for 2 years postoperatively. There were no signs of local recidivation. The same patient had also a small asymptomatic tumour at the other side of the carotid arteries. DISCUSSION: The carotid body tumour originates from the paraganglious tissue at the carotid artery bifurcation. There are angiomatous and adenomatous forms. All of our 6 cases had adenomatous form. It grows slowly, and can compress and/or infiltrate carotid arteries and nerves. Three of our 6 cases showed signs of carotid artery compression and 3 showed infiltration to the carotid arteries. Malignant alteration of this tumour is uncommon. (ABSTRACT T PMID- 9340801 TI - [Liver damage caused by sensitivity to anti-inflammatory agents]. AB - Nonsteroidal antiinflammatory drugs may cause hepatic sensibilization and impairment. The incidence of these changes is low and referential data are scarce and related mainly to reaction which clinically corresponds to hepatic or cholestatic reaction. We present a patient who is sensitive to Diclofenac and has hepatic granuloma which developed after the use of the drug. History, clinical presentation, laboratory results, US examination of the liver and pathohistologic analysis of the hepatic biopsy sample enabled us to diagnose hepatitis with granulomatous impairment. Improvement of symptoms and signs of the disease was spontaneous and without administration of glucocorticoid drugs. PMID- 9340802 TI - [Adrenal gland hemorrhage in neonates--radiologic aspects]. AB - INTRODUCTION: The relatively large adrenal glands of the newborn are vulnerable to mechanical trauma during delivery. Great birth weight, difficult labor, perinatal hypoxia and prematurity are predisposing factors of adrenal haemorrhage. Minor adrenal haemorrhage may not cause symptoms. Massive adrenal haemorrhage is uncommon. Symptoms include anaemia and jaundice associated with a suprarenal mass. In cases with severe blood loss acute shock may develop. In 5 to 10 per cent of cases the haemorrhage is bilateral. Ultrasonography has replaced urography in the diagnosis of this condition demonstrating the site and size of the lesion and allowing an accurate follow-up. Within a month after haemorrhage the blood and necrotic adrenal tissue are resorbed and thin calcification appears at the periphery of the gland. Surgery is necessary if haemorrhagic pseudocyst is large and does not resorb spontaneously. MATERIAL AND METHODS: From 1992 to 1996, five patients with neonatal adrenal haemorrhage were treated at the University Children's Hospital in Belgrade. Two of them were females. All patients were born at term by vaginal delivery. Their birth weights ranged between 3200 and 4050 g. At hospitalization infants were aged from 6 hours to 18 days. The first symptom of adrenal haemorrhage was an abdominal mass in three patients. One of them had laparoschisis with guts and stomach protruding out; the surgeon discovered a mass in the right retroperitoneum during operation. Two patients had jaundice associated with anaemia, and sepsis another two. Ultrasonography was done in all patients. We punctured the haemorrhagic pseudocyst (diameter above 5 cm) in three patients and made cystography. Liquid components of pseudocysts were aspirated and sent to bacteriological and cytological analyses. RESULTS: The diagnosis of adrenal haemorrhage was confirmed by ultrasonography in all patients, demonstrating a right adrenal mass (unilateral in all patients), mostly hypoechoic, which displaced the right kidney. Calcification at the periphery of the pseudocyst appeared in one patient. The adrenal haemorrhage disappeared spontaneously in two patients after two months. An attempt to support the adrenal hemorrhagic pseudocystic regression by puncturing and aspirating its content in three patients was successful in one infant. The patient with laparoschisis died because of sepsis and thrombocytopenia. In a patient the haemorrhagic pseudocyst persisted (6 cm in diameter) and was surgically removed. DISCUSSION: Ultrasound is the method of choice in the diagnosis of adrenal haemorrhage, antenatally and neonatally. It also allows diagnosis of coexisting complications such as renal vein or inferior vena cava thrombosis and a proper follow-up. Puncture of pseudocyst and aspiration of liquid components may support involution of large haemorrhagic pseudocysts. If it is unsuccessful, surgery is necessary. PMID- 9340803 TI - [Health implications of obesity]. AB - The high prevalence of obesity in our society makes obesity a major and increasing health hazard. Extensive observations pointed out a connection of obesity with a variety of illnesses. The evidence shows that obesity is a major contributor to the high prevalence of cardiovascular diseases, diabetes, hypertension, dyslipidaemia, gallstone disease, osteoarthritis and hyperuricaemia in the general population. Obesity also appears to be a contributor to cancers of the colon, prostate and rectum in men and breast, ovary, uterus and biliary tract in women. Hormonal abnormalities tend to occur in both obese men and women. Obese women have menstrual irregularities more often than nonobese. Obese persons tend to have reduced pulmonary function compared with nonobese. Although there are many unresolved issues, weight control is necessary for avoiding and reducing health implications of obesity. More information is required for controlling obesity and its consequences, prior to recommending a dietary advice and treatment. PMID- 9340804 TI - [Evaluation of quality of life in oncology]. AB - Studies of quality of life are more and more becoming an integral part of cancer care in situations when treatment of cancer patients is burdened with significant toxicity, and results of survival of these patients are unsatisfactory. Modern concept of health-related quality of life measurement means consensus about its two basic features: multidimensionality of concept, and essentially subjective experience in a treated person. As a minimum, domains of physical, psychological and social are assessed, and the patient is considered to be the primary source of information. The most widely used instruments for quality of life assessment are questionnaires, especially developed for cancer patients, with supplemental items for particular diagnosis. These instruments must satisfy the basic psychometric properties-validity and reliability. Quality of life studies are focused mostly on phase III trials, where differences between treatment arms for tumour response and survival are expected to be small. Another approach is developed in parallel which, based upon quality of life measurement, initiates decisions for the treatment of a set of patients up to health policy level. In these "cost-utility" studies, efficacy and cost of the treatment, and improvement of patients' quality of life are independently estimated, and then integrated in medical decision making. PMID- 9340805 TI - Control of hemorrhage and elimination of immune inhibitors in a female patient with a severe form of von Willebrand's disease. PMID- 9340807 TI - [Diagnosis and therapy of splenic abscess exemplified by 3 unusual cases]. PMID- 9340806 TI - [Radiologic-pathologic conference. Nodular fasciitis]. PMID- 9340808 TI - [Occupational radiation exposure of assistants in the surgical operating room]. PMID- 9340809 TI - [Spiral CT of pulmonary blood vessels. Diagnostic value of 3D reconstruction]. PMID- 9340810 TI - [New developments in ultrasound diagnosis: 3-dimensional ultrasound]. PMID- 9340811 TI - [Glomus tumors of the petrous bone]. PMID- 9340812 TI - [Advantages of secured nailing]. AB - At the traumatological department of the Surgical Clinic in Prague 5-Motol up to 1986 practically all operations of the skeleton were made by the open method, i.e. under direct visual control. After the Clinic had bought an X-ray apparatus with an amplifier and pulsed skiascopy the number of closed operations increased. The time taken by the operation is shorter and the operations are less risky, in particular for patients of the more advanced age groups. At present the department prefers secured nailing of the tibia, femur and humerus. The introduction of this surgical technique caused a radical change in the surgical approach and therefore the department does not use any longer splint osteosynthesis for diaphyseal fractures of the long bones. PMID- 9340813 TI - [Surgery in the region of the ankle joint--5 years' experience]. AB - During the period January 1992-August 1996 we operated 143 patients with ankles fracture in the department for surgery Decin. An operation technic using the method of transtibial cerclage is presented. The success of the method was evaluated clinically and according to the length of work inability as well. The author emphasizes early rehabilitation in precisely performed osteosynthesis. PMID- 9340814 TI - [Surgical treatment of (selected) calcaneal fractures]. AB - The authors present the short-term results of a prospective group of 23 casualties with 28 intraarticular dislocated fractures of the calcaneus who were treated between November 1994 and May 1996 by open reposition and stabilization by means of a plate, supplemented in 22 cases by autospongioplasty. The most frequent complication as regards healing were infections which were recorded seven times, incl. one case with a deep infection, which penetrated to the bone. By subsequent treatment the infection was in all instances cured, the next follow up period being on average 5.6 months (2-8 months). Metal was extracted in 14 cases, incl. seven cases where it was part of treatment of the infection. For closure of the secondary defect in two instances a microsurgically transferred flap was needed. The patients were followed-up on average for 21 months after the primary operation (6-23 months). A total of 26 calcanei from 21 patients were evaluated. The authors compare two evaluation patterns. According to the authors in 16 instances an excellent result was achieved, a good result in 7 and a poor result in 3 patients. The authors are of the opinion that intraarticular fractures of the calcaneus should be treated by open reposition and stable synthesis. It is not necessary to operate non-dislocated and minimally dislocated fractures, while grossly comminutive fractures should be primarily indicated for arthrodesis. The surgeon must be prepared for the treatment of complications incl. a microsurgical solution of secondary defects. PMID- 9340815 TI - [External fixation in the foot and distal portion of the leg]. AB - The author focused attention on the possibility of external fixation of the foot and distal leg. He uses an external device-MIDI. The use of this type of fixation device is suitable in all types of fractures and dislocations in the region of the foot and distal leg and reduces the period of treatment with variable use of free movement in the ankle joint or free movement of the toes after application of the external fixation device. PMID- 9340816 TI - [Resection of the first rib for the upper thoracic outlet syndrome-- long-term experience]. AB - The authors describe their own experience with the surgical treatment of the upper thoracic aperture syndrome-TOS-during the period of 14 years. During this period on account of this diagnosis at the Surgical Clinic in Brno 112 patients were operated, whereby the resection of the first rib was always done by the transaxillary approach. The authors describe in detail the symptomatology, diagnosis and surgical treatment. They emphasize the advantages of the transaxillary approach. In the conclusion the authors discuss the fact that only in 45% of the operated patients the symptoms disappeared. This leads to reflections on more strict criteria for the surgical treatment of this syndrome. PMID- 9340818 TI - [Plication of the diaphragm--a method of surgical treatment of diaphragmatic paralysis in neonates and infants after heart surgery]. AB - The cause of paresis of the diaphragm after cardiosurgery is damage of the phrenic nerve. The diagnosis of paresis is based on X-ray examination, sonography and electromyography of the diaphragm. Plication of the diaphragm is indicated only in those children with paresis of the diaphragm who develop during spontaneous ventilation severe respiratory insufficiency. In the Cardiocentre of the Faculty Hospital Prague-Motol between 1983 and 1996 of 5333 children operated on account of heart disease 29 children were subjected to plication of the diaphragm, incl. five where the operation was made during the neonatal stage (17%), 20 in infant age (69%) and four were older than one year (14%). By the third day after plication 9 children (38%) could be disconnected from the respirator, by the 5th day 20 children (70%) by the 7th day 22 children (75%). In neonates and infants with postoperative paresis of the diaphragm, where spontaneous ventilation cannot be induced, plication of the diaphragm is according to the authors the method of choice. It is a rapid and safe surgical operation which reduces the period of artificial ventilation and its complications. PMID- 9340817 TI - [A rare cause of ileus and its laparoscopic treatment]. AB - Stenting is a modern method making possible to open malignant oesophageal stenosis. The technique is associated with various complications. The authors describe the management of a rare stent migration to the aboral part of the GI tract, causing an acute abdomen. Based on their own experience with dislocated stents they recommend, contrary to the literature, an active approach when migration to the aboral of the GI tract occurs. Early stent extraction even when the course is uneventful is advantageous. PMID- 9340819 TI - [Long-term results of femorocrural arterial reconstruction]. AB - The objective of the paper is the fate of femorocrural arterial reconstructions during the long-term follow-up of 197 patients operated in 1970-1993. The patients were followed up after operation at 3-12-month intervals and cumulative curves of the patency of the reconstruction and preservation of the extremity were assessed by Kaplan-Meier s method. It was revealed that 5-year cumulative patency of femorocrural reconstructions is 55% and five-year preservation of the extremity even 78%, regardless of the patency of the reconstruction. The authors did not find a difference in the cumulative curves of patency between different crural arteries. The arteria fibularis is from the aspect of preservation of the extremity equivalent to both other arteries. From the comparative result of curves of cumulative patency for the reverse autologous asphena and for cast vascular prostheses ensued that even after five years there is no substantial difference between the two types of prostheses. In case of cast prostheses we must however foresee early thrombotic obstructions and thus repeated surgery such as thrombectomies sand plastic operations of distal anastomoses. PMID- 9340820 TI - [Present possibilities and the role of imaging methods in the diagnosis of acute abdomen, particularly ileus]. AB - The authors compare the importance of modern imaging methods and simple X-rays of the abdomen for the accurate diagnosis of ileus conditions. They discuss the possibilities of different methods and emphasize the importance of series of simple X-rays of the abdomen in different positions. The objective of the recommended procedures is to examine as well as possible a group of patients with an obstruction in the small or large intestine, to localize the obstruction, assess its etiology and not merely state that on the simple X-ray the horizontal beam reveals levels. PMID- 9340821 TI - [One day surgery--personal experience]. AB - The authors present their own experience with surgery carried out in the out patient department in 1985-1996. It is a modern, effective, reliable and above all economical therapeutic method, because it maintains the standard of surgery while saving the costs of hospitalisation. It is a very attractive method for patients who want to return to work as soon as possible. As many as 87% patients were satisfied with surgical treatment of the anus and rectum. PMID- 9340822 TI - [Personal observations on the traumatology system in Israel]. PMID- 9340823 TI - [VATS in pulmonary lobectomy or pneumonectomy]. PMID- 9340824 TI - [Significance of monitoring cytokine levels in the prognosis of acute pancreatitis]. AB - The importance of cytokines as mediators and immunomodulators is increasing in all disciplines. Acute pancreatitis is a disease where the trigger mechanisms of the inflammation and thus also the influence of cytokines are very closely associated and are therefore apparent. The authors investigated on that model the levels of IL-2, IL-6, IL-8, IL-2r and neopterin in relation to the stage of the disease and time. They provided evidence in a group of 33 patients that the cytokine level indicates very accurately the stage of the disease and thus also its prognosis. For the assessment proper of the diagnosis the cytokine level is less important as the rise of the level is quite unspecific. PMID- 9340825 TI - [Antibiotic prophylaxis in thoracic surgery]. AB - On a group of patients operated during the last three months under standard precautions the authors explain their views as regards antibiotic prophylaxis in thoracic surgery. They justify not only "short-term prophylactic administration of antibiotics" but also prolonged prophylaxis with antibiotics. Useful in this respect proved Mandol, cefalosporin of the 2nd generation, which is used as a standard drug without preoperative examination of the bacterial flora in sputum. Their patients did not develop infectious complications. PMID- 9340826 TI - [Aneurysm of an anomalous common celiac-mesenteric trunk]. AB - The authors describe the successful surgical treatment of a very rare aneurysm of the common coeliacomesenteric trunk in a 68-year-old woman. They mention the symptoms, diagnostic procedure and possibilities of surgical treatment of aneurysms of visceral branches of the abdominal aorta. PMID- 9340827 TI - [Incarcerated scrotal hernia containing a sigmoid colon carcinoma]. AB - The authors describe the case-history of a 65-year-old patient with an incarcerated scrotal hernia on the left side. An unexpected finding in the sac of the scrotal hernia was the sigmoid colon with an obturating tumour. The patient was subjected to two-stage surgery with a favourable final effect. PMID- 9340828 TI - [Initial experience with angioscopy in infrainguinal reconstruction using the in situ saphenous vein technique]. AB - The authors report on their experience with lower limb revascularization using the in-situ saphenous vein bypass grafting in four patients with angioscopically assisted valvulotomy. They describe the surgical technique employed and outcome of the procedure. The discussion section examines the pros and cons of the technique of in-situ saphenous vein bypass with angioscopically assisted valvulotomy (ISB + AV) compared with the standard technique of reversed bypass (RVB). PMID- 9340829 TI - [Mesenteric venous thrombosis probably due to hereditary thrombophilia in an 18 year-old boy]. AB - The authors describe the case-history of an 18-year-old patient with an extensive venous mesenteric thrombosis. The case proved fatal despite repeated surgery as a result of relapsing gangrene of the small intestine with diffuse stercoral peritonitis. Thrombosis of the mesenteric veins is a rare disease and accounts only for 4 to 10% of all acute intestinal episodes. The cause of the disease is either idiopathic but more frequently it is associated with various types of coagulopathy. In this context the authors discuss etiological factors, symptoms, diagnosis as well as possible treatment of acute mesenteric venous thrombosis. PMID- 9340830 TI - [Laparoscopic inguinal hernioplasty--results in the initial group 2 years after surgery]. AB - The authors evaluate retrospectively after a two-year interval the results of laparoscopic inguinal hernioplasty (L.I.H.) in an initial group of 49 patients (52 hernias) operated by three surgeons using the method of transabdominal preperitoneal plastic operation (TAPP) in 1993, 1994 and 1995. They confirmed the expected favourable results, except for the very high incidence of relapses (6.7%). In view of the small number of patients other conclusions cannot be drawn except analyse the three early failures of L.I.H., indicate technical mistakes made when introducing the new operation and emphasize the necessity to adhere to the known basic technical details when performing this relatively pretentious laparoscopic operation. PMID- 9340831 TI - [Iatrogenic lesions of the ureter in women]. AB - The changing etiology of ureteral lesions is associated with the expanding spectrum of ureteroscopic, gynaecological, coloproctological and vascular operations. During 1989-1995 the authors treated 60 women with a ureteral lesion, mostly in conjunction with gynaecological operations (37 patients). The assessment of the diagnosis was sometimes delayed. The most frequent initial treatment was the establishment of percutaneous puncture nephrostomy (in 47 instances before delayed reconstruction). The authors discuss the time interval between the development of the ureteral lesion and its reconstruction. The most frequent treatment was ureterorhaphy, ureterocystoneoanastomosis or Boari's flap. PMID- 9340832 TI - [Vesicovaginal and urethrovaginal fistulae]. AB - The authors discuss treatment of vesicovaginal fistulae (VVF) in 36 patients and in six patients urethrovaginal fistulae treated in 1989-1995. The most frequent cause of VVF were iatrogenic lesions after hysterectomy. Occlusion of the fistula was performed 12x by the transvesical approach, nine times by a combined transperitoneal and vaginal approach, four times by a transvesical and transperitoneal approach, eight times by the vaginal route only and three times the authors had to make a continent derivation of urine of the sigma-rectum "pouch" type. Continence by primary operation was achieved in 86%, in urethrovaginal fistulae one reoperation was necessary. With the development of radical operations in the lesser pelvis in women the incidence of iatrogenic lesions is rising slightly, however when the technique of minimal invasive reconstruction urology is used, the prognosis of occlusion of fistulae is favourable. PMID- 9340833 TI - [Pregnancy after urinary bladder augmentation]. AB - Pregnancy after augmentation enterocystoplasty is an increasingly frequent phenomenon in women with congenital anomalies or a neurogenic urinary bladder. Possible complications during pregnancy and the period near delivery and their treatment is described by the authors, based on the case-history of a 22-year-old primipara after ileocoecal augmentation. The authors summarise the so far scarce data reported in the literature. PMID- 9340834 TI - [Should HIV-infected students be allowed to continue their studies?]. PMID- 9340836 TI - [Genetic predisposition and radiation sensitivity of normal tissue]. AB - BACKGROUND: After radiotherapy there are always some patients who develop strong acute and late reactions in normal tissues. In these patients frequently a genetic predisposition is observed. There are found DNA-repair deficiencies and changes in the regulation of the cell cycle which are responsible for the increased radiosensitivity with enhanced cell killing. METHODS: The micronucleus test and the comet assay appear to be appropriate tests in order to measure this increased radiosensitivity. Both tests are characterized by being relatively quick and simple and can be performed with small cell numbers. It is possible to study blood lymphocytes and fibroblasts with these tests. RESULTS: Both tests can predict the radiosensitivity of normal tissues especially if they are applied in combination. CONCLUSIONS: Epidemiological studies with patients after radiotherapy show evidence that the increased radiosensitivity also causes an enhanced induction of secondary tumors by ionizing radiation. This is supported by corresponding animal models. PMID- 9340838 TI - [New approach to conservative therapy of radiation injuries]. AB - PURPOSE: The improvement of tumor control rates and improved survival times of radiotherapy patients result in an increasing importance of radiation side effects in normal tissues. METHODS: Possibilities for the modulation of normal tissue reaction by stimulation of tissue regeneration, or by interference with general pathogenetic pathways which are not specific for radiation damage, are illustrated by a number of examples. RESULTS AND CONCLUSIONS: Increasing knowledge about the pathogenesis of normal tissue radiation responses are expected to significantly improve the efficacy of prophylactic means and possibilities for conservative management of side effects of radiation therapy. Novel approaches may be developed if the so-called "humoral radiation pathology" is taken into consideration in addition to the cellular effects of radiation. PMID- 9340837 TI - [Genetic predisposition and radiation sensitivity of tumors]. AB - BACKGROUND: Studies on normal tissue radiation sensitivity have demonstrated profound differences of individual sensitivities. A number of genetic syndromes associated with abnormal radiation sensitivity have been described. Significant differences have also been detected in persons without known genetic disorders. The question arises as to whether tumors originating from normal tissues with abnormal radiation sensitivity share this abnormal sensitivity and as to whether a general correlation between normal tissue sensitivity and tumor tissue sensitivity can be substantiated. METHODS: Experimental and clinical data derived from own investigations and an extensive review of the literature was used to answer the question. RESULTS: Experimental studies on normal and tumor tissues of SCID-and C3H-mice demonstrated that the 2.7-fold enhanced radiation sensitivity of SCID normal tissues is also found in SCID tumors. Clinical investigations on cervical carcinoma and breast cancer patients revealed higher local control rates in patients with more pronounced acute side effects. A weak trend towards the same relationship was found in head and neck cancer patients. Case reports on unusually severe acute radiation side effects or unexpected tumor remissions as well as few reports on radiotherapy in ataxia telangiectasia (AT) patients suggest a correlation between normal- and tumor-tissue radiation sensitivity. Studies on fibroblasts and tumor cells from the same patient support this hypothesis in soft tissue sarcoma patients, but do not so for head and neck cancer patients. Tumor cells exhibit a considerably higher variation of radiation sensitivities than normal tissue cells. CONCLUSIONS: Experimental and clinical data are compatible with the hypothesis that normal tissue radiation sensitivity predicts for tumor tissue sensitivity. However, in view of the larger heterogeneity of tumor cell radiation sensitivity as compared to normal tissue radiation sensitivity, the development of a clinically useful predictive test for tumor sensitivity based on normal cell sensitivity appears to be unrealistic. PMID- 9340839 TI - [Proliferation rate and radiosensitivity: were Bergonie and Tribondeau wrong?]. PMID- 9340840 TI - [Value of adjuvant progestagen therapy in patients with endometrial carcinoma]. PMID- 9340835 TI - [Evaluation of cancer risk through genetic analysis?]. AB - BACKGROUND: The recent literature of familial cancer, specifically related to germline mutations of RB1, p53, NF1, ATM, BRCA1, Mismatch repair genes and APC is reviewed. RESULTS AND CONCLUSIONS: Germline mutations do not relate to an increased tumor risk of any single tissue, but instead to spectra of neoplastic diseases. The genetic background plays a major role in modifying the cancer risk. Therefore, mass screening for mutations of single genes seems to be without practical value. Only in combination with an adequate and informative family history can molecular genetic analysis significantly support the care for the individual. Comparison of the data of patients inheriting germline mutations and the experience from the corresponding "knockout" mouse demonstrate that only the p53 and APC knockout mice are useful models of human disease. PMID- 9340841 TI - [Multimodal therapy or surgery alone in adenocarcinoma of the esophagus?]. PMID- 9340842 TI - [Reduction of local recurrence and distant metastases in advanced rectal carcinoma by preoperative radiotherapy--results of a randomized study by the MRC (Medical Research Council)]. PMID- 9340843 TI - [Silence is foolish]. PMID- 9340844 TI - [Should physicians be punished?]. PMID- 9340845 TI - [Emergency service in primary health care]. PMID- 9340846 TI - [Hyperkinetic disorders--also in adults]. PMID- 9340847 TI - [Dermatology in practice--practical dermatology]. PMID- 9340848 TI - [Quality assurance of psychiatric work with autistic children and adolescents. Diagnostic value of medical examination]. AB - The literature on associated medical diseases in autism is contradictory and so are the guidelines for medical routine screening. Recommendations draw on epidemiological and population-based research. It was necessary to know the diagnostic yield from patient groups referred to psychiatric clinics. 49 autistic probands were selected from a large clinic pool referred to the Department of Child Psychiatry, Haukeland Sykehus, University Hospital in Bergen, Norway, over the 25 years 1970 to 1995. Detailed analyses were performed regarding referring agent, family history, perinatal data, medical and developmental history, psychometric data, and clinical, neurological and laboratory examination. Our clinical sample deviated from the accepted characteristics of autism: All except one (98%) were mentally retarded. Yet tuberous sclerosis, fragile X syndrome and other known medical disorders said to be associated with autism were not found. More common medical disturbances were found regularly also in those with a higher level of functioning. The likelihood that our blind screening, with comprehensive laboratory examination, would yield positive results was negligible. These clinically important differences, together with a unique make-up of developmental deviances and delays, necessitated individually tailored assessment and treatment in most cases. PMID- 9340849 TI - [Traditional surgical treatment of choledocholithiasis. An analysis of a 10-year material 1980-89]. AB - During 1980-89, 224 patients, 129 women and 95 men, median age 72 years (18-96 years), were treated for common bile duct stones. 26 of the patients had remote cholecystectomy. 67 patients had additional acute cholecystitis, 37 acute cholangitis and 25 acute pancreatitis. 173 patients underwent a traditional open operation, 37 endoscopic papillotomy (EPT) and 14 were treated conservatively. No deaths occurred after elective operations in 52 patients, and one death occurred after early planned operation in 95 patients. Emergency operations and delayed operations for acute disease were encumbered with a lethality of 12%. During the last two years of the study, old septic patients were treated with papillotomy, and there was no mortality among the last 39 patients. The study shows that non septic patients with common bile duct stones can be safely treated by open operation. Old patients with severe complicated gall stone disease should be treated by endoscopic papillotomy at an early stage. PMID- 9340850 TI - [Early elective cholecystectomy in acute stone-related cholecystitis]. AB - During the period 1980 to 1989, 342 patients with acute cholecystitis, 202 women and 140 men, with median age 71 (19-100) years, were admitted to our department. The treatment strategy during the period was early planned cholecystectomy in operable stabile patients with a duration of the disease of less than 7-8 days. Seven patients (2.0%) died, three after emergency operation, three after delayed operation when conservative treatment had failed, and one after medical treatment only. None of 192 patients treated with early planned operation died, and there was no lethality among the patients below the age of 75. The stay in hospital was reduced by 5.2 days after early planned operation. Early planned cholecystectomy for acute cholecystitis is a safe and cost-effective treatment. PMID- 9340852 TI - [Paraneoplastic cerebellar degeneration. A case report]. AB - Paraneoplastic cerebellar degeneration is a rare remote effect of ovarian and breast carcinoma especially, and is characterised clinically by rapidly evolving pancerebellar symptoms. A woman aged 83 developed progressive vertigo, cerebellar ataxia, nystagmus and dysarthria. The cerebrospinal fluid showed slight mononuclear pleocytosis, elevated total protein and IgG concentrations, and oligoclonal bands. A magnetic resonance investigation performed within the first month of symptoms was normal. A left pelvic mass was found, possibly a carcinoma of the colon or the left ovary. Cancer antigen 125 was elevated in the serum and antibodies against Purkinje cells (anti-Yo antibodies) were demonstrated in the serum and cerebrospinal fluid. These results suggested a carcinoma of the ovary as primary site of cancer. Autopsy revealed a left ovarian adenocarcinoma and marked loss of Purkinje cells in the cerebellum. The case illustrates that anti Yo antibodies may serve as a marker not only for paraneoplastic cerebellar degeneration, but also for the nature of the neoplasm that caused it. PMID- 9340851 TI - [Can mortality in gallstone disease be reduced? An analysis of 1013 patients]. AB - A 10-year retrospective review of 1,013 patients with gallstone disease is analysed. The median age of the patients was 66 (18-100) years. 499 patients (49%) were admitted as emergencies. There was a significant relationship between the patient's age, complicated disease and lethality. The mortality was 1%. No patients below the age of 70 died. There was also a significant relationship between duration of the disease and mortality. Emergency operations and delayed operations for acute disease were encumbered with the highest lethality (7%), while early planned operation for acute disease and elective operations showed a lethality of 0.5 and 0.2% respectively. We advocate a more liberal attitude towards elective operations and early operative intervention in elderly patients who do not respond to medical treatment. PMID- 9340853 TI - [Attention deficit/hyperactivity. Diagnostic criteria and classification]. AB - General problems regarding diagnostic criteria and classification of attention deficit/hyperactivity disorder are discussed. A comparison of the most recent versions of the present diagnostic systems shows that they have converged and now overlap each other considerably. ICD-10 (clinical criteria) is still descriptive. DCR-10 (ICD-10's criteria for research) and DSM-IV both have operational criteria. The symptom lists of DCR-10 and DSM-IV are almost identical, and the criteria are very similar. The divisions into types, however, are very different, and none can be recommended. In clinical practice, and particularly in research, it is recommended that cases fulfill the criteria for "disturbance of activity and attention" (F90.0) in DCR-10 or "attention deficit/hyperactivity disorder, combined type" (314.01) in DSM-IV. There are considerable differences between former and present criteria. This must be considered when research results based on former criteria are examined. PMID- 9340854 TI - [Attention deficit/hyperactivity disorder in adults. Diagnostic problems]. AB - Following several reports on treatment of adults with attention deficit/hyperactivity disorder, diagnostic problems regarding the diagnosis are discussed. Symptoms and criteria have changed considerably over the years. Many of those who were given this diagnosis earlier would probably be given a different diagnosis today. Patients should be diagnosed according to the latest diagnostic criteria, but these are developed for children, and are not always adequate for adults. Knowledge regarding the manifestation of the disorder in adults is not sufficient. In order to make this diagnosis in an adult, there must be evidence of the symptoms at the time of examination and also in childhood. The latter may be difficult or impossible to demonstrate. Reliable information on the symptomatology of attention deficit/hyperactivity disorder in adulthood will not be available until children who are given the diagnosis today according to DSM-IV or the research criteria of ICD-10 have been followed up in adulthood. PMID- 9340855 TI - [Organization of epilepsy care for people with mental retardation. A situational analysis after the care reform]. AB - The special care services for the mentally retarded in Norway were closed down by law on 1 January 1991. This implied the termination of centralized care, and integration of these individuals into the local communities. To map the epilepsy service facilities for mentally retarded patients after the care reform, questionnaires were sent to all officially responsible community physicians and to all Departments of Neurology at Norwegian hospitals. 20% of the community physicians expressed the view that mentally retarded patients did not receive adequate specialist service for their epilepsy. There were wide variations within the country. 37% of the responders stated that the community care was inadequately equipped to attend to the special needs of these patients. Firstly, there seems to be a lack of professionals able to transfer their competence concerning epilepsy. Many of the neurological departments felt that the number of referrals of patients with epilepsy and mental retardation had increased since the care reform. 50% of the responders had no opinion about whether the quality of the epilepsy service was not the same for the mentally retarded as for other patients. Many did not realize the need for special measures to provide adequate medical follow-up of this group. It is concluded that education and guidance to personnel and carers should receive high priority in the further development of a comprehensive epilepsy service. The development of multiprofessional epilepsy units at all university hospitals should be facilitated, and the appointment of specially assigned epilepsy nurses at all neurological departments is recommended. PMID- 9340856 TI - [Catheter treatment of congenital heart defects. Arterial septal defects and open ductus arteriosus]. AB - The results of new methods for catheter treatment of congenital heart defects are presented. Between 1989 and 1996 closure of a patent ductus arteriosus was performed in 66 instances on 63 patients, eight of which were with coils. Three patients were treated twice, one with an additional umbrella, two with coils. The overall complete closure rate for umbrellas was 75%, after two ducts, which were initially totally occluded, recanalized. In six more patients the procedure was either aborted or indication was not present. All six ducts treated with coils as the first procedure were completely closed. One of two patients who had residual leak after previous umbrella treatment achieved complete closure after subsequent coil implantation. Closure of atrial septal defects in the oval fossa was performed using the Amplatzer septal occluder in seven children. Complete closure was achieved in all of them. There have been no complications, in particular there have been no cases of embolization in any of the groups. The results seem to indicate that coil occlusion of a persistently patent duct may be at least as good as the umbrella in terms of complete closure. So far both methods have been safe, but experience with coils is limited. The closure of atrial septal defects shows encouraging results. We will continue to offer this treatment as an alternative to open heart surgery in carefully selected patients. PMID- 9340857 TI - [Follow up after potential curative surgery of colorectal cancer. Guidelines from the Norwegian Gastrointestinal Cancer Group]. AB - In Norway, about 2,800 cases of colorectal cancer are diagnosed every year. Two thirds of the patients undergo potentially curative surgery and almost half of them develop local or distant metastases. The follow-up of colorectal cancer patients involves four strategies: Educating the patients about the disease, symptoms of relapse, and risk of hereditariness; Early diagnosis of relapse, to make curative re-surgery possible; Diagnosis of metachronous/synchronous cancer(s); Recording the results of current surgical techniques. The Norwegian Gastrointestinal Cancer Group recommend a four-year follow-up programme (every third month for two years and then twice a year) of colorectal cancer patients. It is suggested that patients treated with low anterior resection are followed regularly by means of rectoscopy and local examination (digital or by ultrasound) undertaken by specialist (surgeon or gastroenterologist). The others should be followed up mainly by general practitioners. Carcinoembryonic antigen (CEA) monitoring is suggested every third month for two years, and then every sixth month. Colonoscopy is recommended at one and four year follow-up. Patients with normal CEA levels prior to surgery should be evaluated by ultrasound of the liver every sixth month for four years. PMID- 9340858 TI - [Forensic autopsy after possible medical malpractice. A 3-year material from the Institute of Forensic Medicine in Oslo, 1993-95]. AB - According to Norwegian law, cases where medical diagnostic procedures or treatment may have caused death shall be reported to the police. During the period 1993-1995, an autopsy was performed at the Institute of Forensic Medicine, Oslo, in 76 cases of such deaths in hospital. In only one case did the police investigation result in a sentence for malpractice. The chief county medical officers react more often; in 16% of the cases, hospital procedures were criticised, or the hospitals were advised to improve hospital routines, and another 14% of the cases were reported to the Norwegian Board of Health. Thus, as expected, the chief county medical officers react more often with criticism than the police do with accusation. The appearance of the police in the hospital often implies a serious conflict of cultures. There are good arguments for specialists in forensic medicine acting as intermediaries between the health service and the judicial system. PMID- 9340859 TI - [Emergency services in Vest-Agder. A survey in 15 municipalities]. AB - Data relating to out-of-hours services in the 15 local councils in Vest-Agder county were gathered through a postal survey. Organisation, the public's accessibility, use of services, communication systems, documentation, medical equipment, doctors' competence, doctors' safety and interaction with other primary care services and secondary health care providers are described. In order to ensure that the public have access to emergency medical care during normal opening hours, there is a need for change in health legislation. Improved daytime access would lessen the pressure on the out-of-hours services. Higher priority should be given by local councils to quality improvement of these services. Some centralization of services to facilities staffed by a doctor and auxiliary personnel is advocated. There should be greater restrictions on night-time house calls than is the case today. Doctors providing out-of-hours services should meet on a regular basis to exchange views and experiences. Greater responsibility should be assumed by local councils, for the continuing education of these doctors. The Norwegian health radio network should be more user-friendly. PMID- 9340861 TI - [Treatment of acne]. AB - Acne is a disfiguring disease, occurring mainly in adolescence. It is a common disorder, affecting above 80% of adolescents in some degree. 50% wish to have treatment for the condition. Acne appears in many forms; from the more common comedonal and papulopustular types to the often devastating cystic and fulminant acne. Some patients develop disfiguring scarring and keloidal reactions, which persist for the rest of their lives. With the treatment facilities available in modern medicine, acne problems can be eliminated through topical and/or systemic treatment options. PMID- 9340860 TI - [Treatment of psoriasis in general practice]. AB - Psoriasis is a chronic skin disease which affects about 2% of the population. Several new treatment modalities have been introduced in recent years. This article focuses on topical treatment of mild and moderate psoriasis that can be successfully treated at home. It is important that general practitioners who treat most of these patients are familiar with the best treatment procedures. Phototherapy and systemic therapy, mainly by dermatologists, are described in brief. PMID- 9340863 TI - [Continuing surgical education]. PMID- 9340862 TI - [Adjuvant treatment in operable colonic and rectal cancer. A new possibility]. PMID- 9340864 TI - [Is birth control counseling free of charge?]. PMID- 9340866 TI - [Law and ethics in occupational medicine]. PMID- 9340865 TI - [The Norwegian fibromyalgia epidemics' growth--and possible decline]. PMID- 9340867 TI - [Explanations of the development of abnormalities in physiological and medical manuscripts in ancient times]. PMID- 9340868 TI - [Laparoscopy in malignancy]. PMID- 9340869 TI - [Hypophyseal tumors: diagnosis and treatment]. PMID- 9340870 TI - [Alzheimer's disease and genes]. AB - Alzheimer's disease is genetically heterogeneous. The rare familial early-onset form of the disease is caused by dominant mutations in at least four different genes. Three of these genes have now been identified and the gene for presenilin 1 (PS1) on chromosome 14 is mutated in about 75% of the families. By contrast, the common form of Alzheimer's disease has late onset and may occur as sporadic cases in the families. This form is multifactorial and the most important genetic risk factor is the E4 allele of the polymorphic apolipoprotein E gene (APOE) on chromosome 19. The E4 allele is associated with moderately or strongly increased lifetime risk of Alzheimer's disease in persons with respectively one or two copies of the gene variant. Apolipoprotein E genotyping may serve as an adjunct in the diagnostic evaluation of Alzheimer's disease, but predictive genotyping of asymptomatic persons is premature and should not be done. PMID- 9340871 TI - [Epidural glucocorticoid injection in lumbago sciatica]. AB - Within the past decades, epidural steroid injections have been used in the treatment of severe low back pain and sciatica. In reviewing papers for this article an effort is made to concentrate on those that meet commonly accepted research design criteria, such as being blinded, randomized and prospective. The risks and the advantages of the procedure are discussed. Some of the studies report an efficient reduction in low back pain and sciatica for a longer period. Risks of more serious complications are low using the right technique. However, the results are to some extent conflicting. Future correctly designed studies are necessary to clarify whether the injection should be a supplement to the established treatment of low back pain and sciatica. PMID- 9340872 TI - [Apolipoprotein E genotypes in patients investigated for dementia]. AB - Certain genotypes of apolipoprotein E (apoE, locus APOE) are associated with an increased risk for development of Alzheimer's disease. We present the distribution of the APOE alleles (E2, E3 and E4) in 50 Danish patients referred to a dementia clinic. The distribution of alleles in patients with dementia was E2: 2, E3: 36 and E4: 38; and in 12 patients without dementia E2: 1, E3: 18, and E4: 5. The frequency of E4 alleles was significantly increased (chi 2 = 42; df = 1; p < 10(-4)) among patients with Alzheimer's disease compared with a Danish control population. The study demonstrates a strong association between Alzheimer's disease and the E4 allele. No difference was found in the frequency of the E4 allele between Alzheimer and non-Alzheimer demented patients. PMID- 9340873 TI - [Contact allergy to local steroids. Contact allergy to corticosteroids among consecutively tested patients with eczema]. AB - We studied the frequency of contact dermatitis to corticosteroids in consecutive eczema patients and the relevance of positive patch test reactions. Of 2,742 patients patch tested at the Dermatology Clinic at Odense University Hospital between June 1991 and December 1995, 65 patients (47 women and 18 men) or 2.4% had a positive reaction to one or more of the corticosteroids tested. Forty (1.45%) had a positive reaction to budesonide 1% pet. in the standard series. For a one-year period 662 consecutive eczema patients were routinely patch tested with a corticosteroid series consisting of five steroid allergens, and 17 had a positive reaction to at least one of these (2.6%). Budesonide allergy was most often encountered, followed by hydrocortisone, tixocortol pivalate and hydrocortisone-17-butyrate. Betamethasone valerate, triamcinolone acetonide and clobetasol propionate rarely caused contact allergy. The corticosteroid allergy was of current relevance in about one third of the cases. Corticosteroid contact allergy is an uncommon adverse effect, in relation to the number of patients with inflammatory skin diseases who are treated with topical corticosteroids. In patients with longstanding eczematous skin disease, who do not improve or deteriorate during topical corticosteroid therapy, contact allergy to these drugs should be suspected and relevant patch tests should be performed to sort out concomitant reactions. Future treatment with corticosteroids must take into account possible cross-reactions. PMID- 9340875 TI - [Endoscopic decompression of carpal tunnel syndrome with the "Agee one-portal system"]. AB - The aim of the present study was to present the results after endoscopic carpal tunnel release in the first 30 hands operated in our department. Over a five month period we used the Agee one-portal system for endoscopic carpal tunnel release in all patients operated with a carpal tunnel release in our department. All patients were operated by one surgeon, who recorded all data and results prospectively. The patients were evaluated preoperatively and after two and eight weeks. A total of 30 hands in 25 patients were operated. There were 19 females and six males with a mean age of 50.1 years (23-80). There were no haematomas, injuries to the arcade vessels, nerve lesions or infections. All patients but one stated a good clinical result of the operation, and all patients had regained normal movement of all fingers at the two-week postoperative check-up. All patients, who were working preoperatively, were able to return to work after two to three weeks except for three patients. In conclusion, we have not experienced any complications in our series, and all patients but one had a good clinical results. We feel that the Agee one-portal system ensures carpal tunnel release with a minimum of discomfort, and we find the preliminary results in our department so promising, that we now use this technique as a routine in uncomplicated cases of carpal tunnel syndrome. PMID- 9340874 TI - [Treatment of acute apoplexy at a medical department with a specialized rehabilitation unit]. AB - The aim of this retrospective study was to describe the occurrence of acute stroke and the effect of treatment measured as mortality, length of hospital stay and discharge to the home in a medical department with a specialized rehabilitation unit. During the period 1.9.1992-31.5.1995 110 patients were discharged to their own home after transient cerebral ischaemia, 23 after subarachnoid haemorrhage, 62 after documented intracerebral haemorrhage and 574 after acute stroke due to infarction or unknown cause. The 636 patients in the last two groups had an in hospital mortality of 18%, a 30-day mortality 18% and a six month mortality of 25%. In the same group the length of hospital stay was 25.6 days and 68% were discharged to their own home. In conclusion the results of treatment of acute stroke in a medical department with a specialized rehabilitation unit were similar to those reported from acute stroke units in Denmark and abroad, but the patients admitted to our department were younger and fewer were single, which may itself reduce mortality and length of hospital stay. PMID- 9340876 TI - [Simultaneous occurrence of trigger fingers and carpal tunnel syndrome]. AB - We report an unusual case of trigger fingers and carpal tunnel syndrome occurring simultaneously; both conditions were caused by space-occupying fibrosis of the flexor tendon sheath. Following operation the patient was relieved of his symptoms. In case of simultaneously occurring trigger fingers and carpal tunnel syndrome, one should be aware that both conditions can be caused by space occupying lesions beneath the carpal tunnel. Endoscopic surgery should be avoided in these cases. PMID- 9340877 TI - [Prognostic factors in patients with disseminated germ cell tumors]. PMID- 9340878 TI - [Percutaneous dilation tracheostomy--a preliminary study]. PMID- 9340879 TI - [Combination preparations of codeine and paracetamol]. PMID- 9340880 TI - [Mortality in cystic fibrosis]. PMID- 9340881 TI - [What is normal?]. PMID- 9340882 TI - [Support and guidelines, nicotine substitutes and clonidine have positive effects on smoking cessation. Five Cochrane meta-analyses on agents used in smoking cessation]. PMID- 9340883 TI - [Cystic fibrosis and chronic Pseudomonas aeruginosa infection. Possibilities for immunologic prophylaxis and therapy]. AB - Cystic fibrosis is an autosomal recessive disease, characterised by chronic pulmonary infections, pancreatic insufficiency and increased electrolyte content of sweat. Cystic fibrosis is diagnosed in one out of 4761 children below the age of 15 years. Pulmonary infection was previously caused by Staphylococcus aureus, but after the introduction of penicillin, the mortality was reduced from 61% to 20% within the first five years of life. Today chronic pulmonary infection is primarily caused by Pseudomonas aeruginosa. P. aeruginosa infection produces an immunologically conditioned destruction of the pulmonary tissue, leading to fatal bronchiectasis. P. aeruginosa develops resistance against most antibiotics and chemotherapeutic agents except colistin. Immunological aspects concerning active and passive immunisation are discussed in the article. Until today no useable vaccine has been found, but several candidates are subjects of research. PMID- 9340884 TI - [Improved prognosis for patients with cystic fibrosis. A result of aggressive center-based treatment]. AB - We report survival data for Danish centre-treated cystic fibrosis (CF) patients, covering the period 1974-1993 using cross-sectional cumulative survival probability based on annual age-specific mortality rates. No significant differences were noted in the survival probability when patients were grouped according to sex or absence/presence of meconium ileus. The annual mortality rate for 1989-1993 was 0-1.2%. Using the age-specific mortality rate for 1989-1993, we were unable to calculate the median survival probability because the curve did not fall below 50% (age up to 45 years). It was, however, possible to show that the survival probability for a CF child born after 1989 to reach his or hers 45th birthday was 80.4% (95% confidence interval 76.5-84.6%). The probability of surviving 40 years after the diagnosis of CF is made was 83.3% (95% confidence interval 80.1%-86.6%). This is considerably higher than any other published survival probability. An aggressive anti-Pseudomonas aeruginosa treatment regimen seemed important in achieving the observed improved survival. PMID- 9340885 TI - [Dementia diagnosis in general practice]. AB - In a follow-up study eight GPs prospectively examined 36 strategically selected patients. As diagnostic tools psychometric tests, blood samples and physical examination including a CT-scan of the brain were used. After one year the surviving patients were reexamined with respect to cognitive testing, ADL function and signs of depression. A panel of experts evaluated the records. The GPs found 19 patients to be demented, five patients possibly demented and five not demented. The experts found 21 demented, three possibly demented and five not demented. After the first examination the GPs and the experts found two patients with possible reversible dementia. However, the patients did not improve concerning their cognitive function despite relevant medical treatment. The degree of dementia was estimated by the GPs as being ten patients with light, ten with moderate and two with severe dementia, while the experts respectively found eight, 13 and two with the same items. PMID- 9340886 TI - [Occupational respiratory tract allergy in trout processing workers]. AB - Out of 16 workers in a trout processing industry, ten experienced work-related cough, dyspnoea, and nasal secretion. A clinical examination was performed including specific IgE, precipitating antibodies IgG for trout and processing water, skin prick testing and peak flow monitoring. A total of four workers showed a positive allergic reaction. Processing water contained endotoxin and bacteria in high amounts. It is concluded, that work-related respiratory symptoms should be investigated and the cause at the workplace identified, so that preventive measures can be introduced. PMID- 9340887 TI - [Intestinal tuberculosis]. AB - Since the symptoms and clinical presentation of intestinal tuberculosis is nonspecific, the diagnosis is frequently delayed and may be achieved at autopsy only. Intestinal tuberculosis is very rare in Denmark, but may now be seen more often because of increasing numbers of immigrants from countries of the third world with a high prevalence of tuberculosis. A case of intestinal tuberculosis in a 28 year old Somalian female is reported. Methods of diagnosing intestinal tuberculosis are commented, and the frequent necessity of starting medical treatment before a bacteriological diagnosis is reached is emphasized. PMID- 9340888 TI - [Dyspepsia in general practice]. PMID- 9340890 TI - [Pressure pneumothorax after intercostal blockade]. PMID- 9340889 TI - [The EU campaign for olive oil is oiled with corrupt science]. PMID- 9340891 TI - [AIDS in Sweden and Denmark]. PMID- 9340892 TI - [Pernicious anemia and cancer risk]. PMID- 9340893 TI - [Antithrombotic treatment in cardiovascular diseases. A report by a working group of the Danish Cardiologic Society and the Danish Society of Clinical Chemistry]. PMID- 9340894 TI - [Reference program om contact eczema. Outlined for the Danish Dermatological Society by the Danish Contact Dermatitis Group]. PMID- 9340896 TI - [Pattern of distribution of constitutive isoforms of NO synthase in the normal prostate and obstructive prostatic hyperplasia]. AB - Nitric oxide (NO) is suggested as an important mediator for the regulation of biological processes. In the present study we tried to determine histochemically and immunohistochemically the localization and distribution of the constitutive NO-synthase Isoforms (bNOS and eNOS) of 14 normal non-obstructive and 12 hyperplastic obstructive human prostates. Differentiated nitrinergic innervation was shown for the prostate glands, fibromuscular stroma and blood vessels by NADPH-diaphorase staining and immunohistochemically with specific NOS antibodies. In the specimens with benign prostatic hyperplasia nitrinergic innervation seems to be distinctly reduced. The vascular distribution of NOS provides evidence for segmental differentiation of NO-mediated vascular regulation. The NADPH diaphorase reaction was not confirmed immunohistochemically by the specific NOS antibody in the glandular epithelium. The distribution of NO synthase shows the importance of nitric oxide in the regulation of smooth muscle tone, blood flow and secretory function in the normal and hyperplastic human prostate. PMID- 9340895 TI - [Occurrence of lymphoceles after allogeneic kidney transplantation and their therapy]. AB - We conducted a prospective evaluation of 301 consecutive renal transplants to define the prevalence and therapy of lymphoceles following renal transplantation. Using a management scheme, we identified in 52 patients (17.3%) perirenal fluid collections. Using this management scheme, we treated 47/52 patients conservatively. Only 5 (1.7%) patients required internal marsupialization. We conclude that groin lympho cysts after renal transplantation can be successfully treated with conservative methods. PMID- 9340897 TI - [Value of PSA density determination in prostate carcinoma]. AB - The PSA density was calculated in 98 patients with carcinoma of the prostate by using the weight and/or volume of the radical prostatectomy specimen. In 22 of 98 patients, PSA serum levels ranged between 4 and 10 ng/ml. Eleven patients showed PSA serum levels below 4 ng/ml and 65 patients above 10 ng/ml. Sixty-one of the 65 patients with PSA serum levels > 10 ng/ml had PSAD values above 0.15, and we therefore suspected the presence of carcinoma of the prostate according to Benson et al. Twelve of the 22 patients with PSA serum levels between 4 and 10 ng/ml had PSAD values below the cut-off value of 0.15. In addition, all 11 patients with PSA values < 4 ng/ml showed PSAD values < 0.15, a range that can be regarded as almost harmless according to Benson et al. In conclusion, it seems apparent that in patients with PSA serum levels < 10 ng/ml, no important additional information is obtained from the PSAD determination. Routine measurement of PSAD for evaluating the risk of carcinoma of the prostate in individual cases can therefore not be recommended. PMID- 9340898 TI - [Combination of Sabal and Urtica extract vs. finasteride in benign prostatic hyperplasia (Aiken stages I to II). Comparison of therapeutic effectiveness in a one year double-blind study]. AB - Therapeutic equivalence should be demonstrated in a randomised, reference controlled multicentric double blind clinical trial with PRO 160/120, a combination of Sabal- and Urtica-Extract, and Finasteride, respectively, in patients suffering from benign prostatic hyperplasia (BPH, Stage I to II according to Aiken). The study involved 543 patients, who were treated for 48 weeks with two capsules of PRO 160/120 or one capsule of Finasteride per day, in a double dummy design. Primary variable was the change of the maximum urinary flow after 24 weeks of therapy in comparison to therapy start. As secondary variables urodynamic parameters such as average urinary flow, miction volume and miction time were monitored. Urinary symptoms were recorded by the International Prostate-Symptom-Score (I-PSS, Paris 1993). Additionally, the impacts of the symptoms on quality of life had been assessed by a quality of life questionnaire according to The American Urological Association Measurement Committee (1991). An increase of the urinary flow rate could be observed in both treatment groups (1.9 ml/s with PRO 160/ 120; 2.4 ml/s with Finasteride). During the trial, the average urinary flow increased, whereas the miction time decreased in both groups in a similar extent. The miction volume did not show any relevant differences after treatment with either PRO 160/120 or Finasteride. The I-PSS decreased from 11.3 at the therapy start to 8.2 after 24 weeks and 6.5 (week 48) under PRO 160/120 and from 11.8 to 8.0 and 6.2, under Finasteride, respectively. Accordingly, life quality improved between therapy start and therapy end from 7.5 to 4.3 with PRO 160/120 and from 7.7 to 4.1 with Finasteride. In terms of safety aspects less adverse events occurred with the Sabal/Urtica-Extract as with Finasteride. Especially less cases of diminished ejaculation volume, erectile dysfunction and headache have been reported. PMID- 9340899 TI - [Is retrograde ureteropyelography necessary before kidney pelvis-plasty in childhood?]. AB - For routine evaluation of ureteropelvic junction obstruction in children, sonography, voiding cystogram, IV pyelogram and a renal scan in combination with administration of furosemide are available. Furthermore, often preoperative antegrade or retrograde ureteropyelography is performed. However, the significance of retrograde ureteropyelography in the world literature remains controversial. Therefore, we reviewed the records of 41 children who underwent a pyeloplasty in our department. In 9 children the ureter was visualized by IV pyelogram, voiding cystogram or antegrade ureteropyelography; a retrograde examination of the ureter was performed in 21 children before pyeloplasty in the OR. In 11 children the ureter was not visualized preoperatively. Retrograde ureterography neither gave additional information in any patient nor did it change the operative technique. In the children where the ureter was not visualized preoperatively, no ureteric abnormality was found in association with ureteropelvic junction obstruction. Hence, we conclude that retrograde ureteropyelography before pyeloplasty in children is not necessary, provided that sonography does not show ureteral dilatation. PMID- 9340900 TI - [Long-term outcome of surgical therapy of metastatic non-seminomatous germ cell tumor in advanced tumor stages]. AB - We present long-term results (minimum follow-up 5 years) in 145 patients with advanced non-seminomatous germ cell tumours, who underwent radical retroperitoneal lymphadenectomy (RPLA) after chemotherapy. We correlated patients' outcomes (death of disease) to different kinds of chemotherapy and to intraoperative findings. We found that patients who were treated by a modified Einhorn scheme with cisplatin, etoposide and bleomycin have a good prognosis. In all, 90% showed no evidence of disease (NED). The NED rate was significantly lower in patients who were treated by sequential alternative chemotherapy (DOD = 37%). We determined the following prognostic factors which predict a poor outcome: salvage RPLA in the case of progressive disease or tumour marker increase during chemotherapy (DOD = 89%, P < 0.0001) residual tumour in multiple organ systems (DOD = 41%, P = 0.0006) vital tumour in RPLA specimen (DOD = 53%, P < 0.0001) residual tumour mass > 5 cm (DOD = 41%, P = 0.0188). We found that histopathological findings of the primary tumour and tumour stage IIc-IIIc according to the Lugano classification have no prognostic significance for death of disease. PMID- 9340901 TI - [Radical prostatectomy as primary monotherapy in capsule penetrating prostatic carcinoma. 15 years outcome]. AB - Twenty-five patients with locally advanced prostate cancer (stage pT3pN0) underwent pelvic lymphadenectomy and radical prostatectomy and were followed up thereafter for at least 15 years. No hormonal treatment was given prior to tumor progression. Overall and disease-free 15-year survival rates were observed to be 44 and 24%, respectively. These data suggest that a cure from prostate cancer by radical prostatectomy can be expected in a quarter of patients with capsular penetration. From our results, no justification can be derived to exclude radical prostatectomy from the spectrum of treatment options for patients with capsular penetration of prostate cancer. More detailed analysis of the results depending on the local extent of the tumor and histological grade revealed distinct differences with respect to the risk of progression. Histological grade was the single most predictive parameter of progression. Out of all subgroups of patients with capsular penetration of prostate cancer, those with a poorly differentiated tumor showed the shortest progression-free interval after surgery, the highest level of overall progression and the largest proportion of tumor-related deaths. By contrast, the prognosis was only slightly influenced by the presence or absence of seminal vesicle involvement. The role of adjuvant treatment after radical prostatectomy for patients with stage pT3pN0 prostate cancer or for subgroups of them remains to be determined within the scope of prospective randomized trials. PMID- 9340902 TI - [Primary retroperitoneal fibrosis with renal involvement]. AB - We report on a patient suffering from primary retroperitoneal fibrosis with histologically proven involvement of the right kidney. The problems in making the correct diagnosis, for which in this case a laparotomy and histological examination were also necessary, are discussed. PMID- 9340903 TI - [Dorsal ligation of the penile vein--prognostic factors and long-term outcome]. AB - Poor longterm success has been reported for penile vein ligation the last few years. Therefore, we decided to re-investigate our group of 147 patients who were operated on between 1987 and 1996. All patients showed a negative response to intracavernous injection therapy at the time of diagnosis and revealed a maintenance flow > 15 ml/min, as well as a pathological venous flow with pharmacocavernosometry or pharmacocavernosography. These patients underwent ligation of all superficial dorsal veins and resection of the deep dorsal vein of the penis. An up-to-date record of the success of the operation was kept either by a renewed clinical visit or by a standardized telephone interview or questionnaire. A total of 126 patients were available here for long-term follow up. We divided the findings into three groups: complete spontaneous erection, postoperative response to cavernous autoinjection therapy and no changes in erectile competency postoperatively. The short-term success rate for these groups after 1-3 months was an outcome of 31 (24.6%), 25 (19.8%) and 70 (55.6%) patients; 86% of the cases whose results deteriorated after the initial operation success rate had this happen within the first postoperative year (p < or = 0.001). Favorable prognostic factors were preoperative erectile dysfunction of < or = 7 years, a normal CC-EMG and a maintenance flow of < or = 45 ml/min. If all three parameters were present, the long-term success rate (spontaneous erection plus response to intracavernous injection) of 30% of all patients was found to rise to 67% in this selected group of patients (p < or = 0.001). This study reveals that long-term success for unselected patients undergoing penile venous surgery is disappointing; however, careful selection of patients by certain prognostic factors can improve long-term results. PMID- 9340904 TI - [Electromyography of the sphincter vesicae externus muscle. Technique, indications and outcome]. AB - Electrophysiological examinations provide important findings for the clarification of urogenital dysfunctions. A routine electromyogram (EMG) of the external anal sphincter is used together with stimulation of the pudendal nerve to determine lesions of the somatomotor tract. An alternative is the electromyographical examination of the external vesical sphincter. This special method requires more time and sufficient experience. We see an indication for EMG of the voluntary vesical sphincter in urogenital dysfunctions that cannot be sufficiently differentiated with EMG of the external anal sphincter and also in disturbances that selectively affect the external vesical sphincter. PMID- 9340905 TI - [Interdisciplinary consensus conference on "diagnosis and therapy of testicular tumors"]. PMID- 9340906 TI - [Primary renal plasmacytoma. A case report]. AB - We describe a patient with a primary renal plasmacytoma. In the literature available, only six clinical cases have been reported. A 64-year-old patient is presented who had the clinical signs of a renal cell carcinoma. Histological examination after nephrectomy, however, revealed a plasmacytoma. Based on this case, we discuss how one should proceed in this disease. Primary renal plasmacytoma can be tentatively diagnosed preoperatively only in the presence of paraproteinemia or Bence-Jones proteinuria, as it cannot be distinguished from other renal tumors by imaging procedures. Postoperatively, further staging procedures must rule out bone involvement (solitary myeloma or multiple myeloma). Nephrectomy is required in patients with plasmacytoma only if renal complications occur. PMID- 9340908 TI - [Supplementary accident insurance]. PMID- 9340907 TI - [Diagnosis and therapy of testicular tumors]. PMID- 9340909 TI - [Ethics and insurance medicine]. PMID- 9340910 TI - [Criteria for evaluating free will in suicide]. AB - Site-related categories of suicide enable certain characteristics to be worked out which speak both for, as well as against, a voluntary decision, and can generally be viewed as indicative. However, taking other site-unrelated factors into account, there are necessarily differences which can only be conditional to the particular case in question. PMID- 9340911 TI - [Gunshot wounds--homicide, suicide or accident?]. AB - Gunshot residues on the hand are an important evidence of shooting. Different methods for sampling gunshot residues from the hands are presented and critically analyzed. The topographic methods-adhesive film and polyvinyl alcohol technique (PVAL)-have serious advantages over the cumulative methods like tape-lift or cotton swab. Case examples demonstrate the limited interpretation of results with cumulative methods. Negative results of tape-lifts or cotton swabs can also be explained by the presence of blood or dirt, positive results however don't prove shooting. The sampling method with the highest gain of gunshot residues is the PVAL technique which takes about 2 hours of sampling traces. Therefore PVAL is mostly applied post mortem. PVAL shows the distribution of gunshot residues also on bloody hands and is a powerful instrument for the reconstruction of the firing position. But the final differentiation between homicide, suicide or accident is not only based on gunshot residues, but requires a complex analysis of all findings in a case (scene, blood traces, autopsy, shot range). PMID- 9340912 TI - [Minimum requirements in medical expert assessment of occupational disability of patients with chronic nonspecific pain]. AB - Assessing working capacity in patients with chronic nonspecific pain disorders applying for disability pensions is a core tasks of practical social medicine. Traditionally the evaluation focused on the nosological classification and clinical description of the pain disorder with rather vague concepts of its consequences in terms of disabilities and handicaps. The author proposes to reverse the sequence. Following the International Classification of Impairments, Disabilities and Handicaps (WHO 1980) work incapacity can be seen as an "occupational handicap". Its qualities, severity and credibility depend on the existence of objectifyable disabilities (especially behaviour, locomotor, body disposition, dexterity and situational disabilities). These may be attributed to clinical impairments and an underlying disease process. Among impairments pain has to be assessed multidimensionally. As in clinical medicine a nosological diagnosis is a useful but neither necessary nor sufficient prerequisite for a sociomedical evaluation of pain patients. PMID- 9340913 TI - [Effect of early diagnosis and therapy on prognosis of Parkinson syndrome]. AB - Parkinson's disease is a chronic-progressive neuro-degenerative syndrome, which should be diagnosed as early as possible because of the therapeutic success in the beginning. Knowing the initial symptoms and using modern technical devices like SPECT and PET we shall be able to influence the course and the prognosis of the disease positively. The application of a current combined drug-therapy and in addition physiotherapy, speech-therapy and psychotherapy enable us to provide quality of life or to restore it. Repeated treatments in Parkinson medical centers intensify the efficacy of therapeutic efforts. Nevertheless even in early stages patients and physicians can be confronted with social medical problems like defining the degree of disability or the time for receiving a pension. The knowledge of specific disease-related findings will help to judge all these problems in accordance with the needs of the patients. PMID- 9340914 TI - [Risk of hepatitis C virus infection in medical occupations]. AB - Following the introduction of sensitive and specific tests for the detection of hepatitis C virus (HCV) infection studies could be performed to determine the risk to acquire a HCV infection in medical professions. The aim of this report is to summarize the knowledge in this field, especially regarding the risk of transmission of HC virus by injuries with infected material, such as needles. The risk to acquire a HCV infection following needlestick injuries was found to be markedly lower than the risk for transmission of hepatitis B virus infection. In 7 studies from different countries this risk has been found to be between zero to a maximum of 10% (average 2%). When the frequency of HCV infection was studied in occupational groups in hospital (nurses, technical assistants, doctors), it did not differ significantly compared to the prevalence in general population. These and other findings concerning the risk of transmission of HCV infections (between spouses, mother/newborns) clearly show that the risk of transmission of HCV infection is markedly lower than that of HBV infection. These facts should be considered in judging of applications for the recognition of HCV infection as occupational disease. PMID- 9340915 TI - [Comment on O. Huth: Accident incidence at various ages]. PMID- 9340916 TI - [Comment on B. Madea and P. Schmidt: Fatal post-traumatic alcohol delirium--right to compensation in private accident insurance]. PMID- 9340917 TI - [Comment on F. Schultz, K. Lieske: Traumatic splenic rupture--fatal thrombocyte increase after splenectomy]. PMID- 9340918 TI - [Multi-drug management after myocardial infarct]. AB - Since the results of studies done with angiotensin-converting-enzyme(ACE) inhibitors, systemic thrombolysis and primary percutaneous transluminal angioplasty (PTCA) have been published, post-myocardial infarction therapy has changed dramatically. In most european countries systemic thrombolysis as early as possible is the standard therapy. Cardiologists prefer more and more immediate revascularisation if the technical standard of the hospital is allowing an acute intervention. Pharmacological therapy is done initially with beta-blockers and platelet aggregation inhibitors, if necessary intravenously, since the results of the e.g. GUSTO- and the AIRE-study are known. If left ventricular dysfunction exists or develops, ACE-inhibitors are given after the third day post myocardial infarction. PMID- 9340919 TI - [Acetylsalicylic acid after myocardial infarct]. AB - There is no doubt about the positive influence of acetylsalicylic acid on vascular disease. The antithrombotic therapy with acetylsalicylic acid is a fundamental of secondary prevention of vascular events, especially in nonfatal myocardial infarction, instable angina pectoris, stroke and in patients with amaurosis fugax. Clinical studies done since 1990 about the effect of a low-dose therapy confirmed the experimental results, which showed that 25-50 mg acetylsalicylic acid per day are enough, to suppress platelet function as far as necessary, without influencing the protective function of prostacyclin synthesis. Side-effects can be reduced to a minimum with a reduced dosage. The wide-spread use of a low-dose acetylsalicylic acid therapy in primary prevention especially in patients at high risk has to be reevaluated in further studies. PMID- 9340920 TI - [Reducing cholesterol in cardiovascular rehabilitation--exercise versus drug therapy]. AB - Prospective epidemiological studies have proven a close link between the improvement of the lipoprotein profile and reduction of coronary heart mortality in coronary heart disease (CHD) patients. Both, regular physical activity and pharmacological intervention are capable of improving the lipoprotein profile and may independently reduce coronary stenoses and cardiac events in CHD patients. Because of the multifactorial genesis of atherosclerosis, regular physical activity is of major importance for cardiac rehabilitation and improvements of other risk factors besides the lipoprotein profile e.g. rheology, hypercoagulability, hypertension, peripheral insulin resistance and oxidative status may be expected. The combination of life-style changes e.g. physical activity, diet and smoking cessation together with pharmacological intervention, if risk factors remain high, seems to be the best therapy for patients with CHD. A sole pharmacological intervention seems to be insufficient. PMID- 9340921 TI - [Current status of drug therapy in rehabilitation after myocardial infarct]. PMID- 9340922 TI - [Significance of supraphysiologic administration of magnesium after myocardial infarct]. AB - Magnesium is an essential element for the regulation of energy dependent metabolism and transmembranous electrolyte flow. Despite the present controversial interest, magnesium has some beneficial effects on several manifestations of coronary artery disease and myocardial infarction. In supraphysiologic serum levels magnesium causes dilatation of epicardial coronary arteries and modulates better diastolic relaxation of the left ventricle. It protects the ischaemic myocardium against calcium overload, preserves the intracellular ATP and brakes the acidosis caused by anaerobic metabolism and free radicals with subsequent cell necrosis. Spontaneous depolarization of the myocardial cell decreases after magnesium administration. Supraventricular and ventricular arrhythmias become less frequent under supraphysiologic magnesium serum levels. Also magnesium inhibits platelet aggregation. The conflicting results of randomized trials examining the efficacy of magnesium supplementation have failed to establish conclusively whether magnesium is generally useful in coronary artery disease and after myocardial infarction. Although findings of experimental studies showed beneficial effects of supraphysiologic administration of magnesium before or in the early phase of ischaemia and reperfusion during acute myocardial infarction. By the way supraphysiologic magnesium application is safe, cheap and has no side effects in common dosage. PMID- 9340923 TI - [Hormone replacement in postmenopausal women with coronary heart disease. Results and clinical consequences from recent studies]. AB - Despite the comparable risk profile, women in reproductive age have less frequently cardiovascular disease than men. Hormone replacement therapy in postmenopausal women is associated with further reduction of coronary artery disease and myocardial infarction. There is good evidence, that estrogens cause these differences. Several well observed biological effects of estrogens seem to be responsible for this benefit. Estrogen replacement therapy reduces cholesterol and LDL-cholesterol levels and has also some antioxidant potential. Furthermore estrogens have effects on vasodilation of the coronary arteries. A benefit on haemostasis through fibrinogen attenuation and platelet inhibition is also described. A main problem of patients compliance during estrogen replacement therapy are postmenopausal bleeding and being afraid of breast cancer. Hormone replacement therapy is indeed not without any risk. The incidence of breast cancer is controversially discussed. Thrombosis and pulmonary infarction are described as further risks. Comparing the risks and benefits of estrogen replacement therapy concerning the reduction of cardiovascular disease, actually the objections are to be dissipated from an epidemiological point of view. Thus if the results of the ongoing large randomized studies prove, that estrogen replacement therapy reduces the risk of coronary artery disease, a challenge in primary- and secondary prevention of cardiovascular disease in women will have to occur. PMID- 9340924 TI - [Visual rehabilitation: magnifying vision aids]. AB - Many retinal diseases can be treated either with laser treatment or with vitreo retinal surgery. However, in some of these cases a severe visual loss cannot be avoided and visual rehabilitation has to be initiated. Visual rehabilitation is initially performed by fitting low vision aids. With low vision aids the patients ability to read can be restored as well as many other needs of every days life e.g. reading bus numbers, street names, bank notes or letters. In analogy with the higher life expectancy in the middle european countries the incidence of age related maculopathy will increase in the next years. Thus, the need for visual rehabilitation will increase as well. PMID- 9340925 TI - [Computer eyeglasses--aspects of a confusing topic]. AB - With the coming into force of the new Austrian Employee Protection Act the issue of the so called "computer glasses" will also gain added importance in our country. Such glasses have been defined as vision aids to be exclusively used for the work on computer monitors and include single-vision glasses solely intended for reading computer screen, glasses with bifocal lenses for reading computer screen and hard-copy documents as well as those with varifocal lenses featuring a thickened central section. There is still a considerable controversy among those concerned as to who will bear the costs for such glasses--most likely it will be the employer. Prescription of such vision aids will be exclusively restricted to ophthalmologists, based on a thorough ophthalmological examination under adequate consideration of the specific working environment and the workplace requirements of the individual employee concerned. PMID- 9340926 TI - [Contact lenses--an overview]. AB - Contact lenses are a modern possibility to correct defective sights as an alternative to glasses. They are not only used because of cosmetic reasons but also if there is optical or therapeutical indication from the medical point of view. For keratoconus and cornea globosa are hard contact lenses not only the unique efficient possibility for correction but also contemporary therapy. If cornea is irregular-for instance when there are cicatrices-an essential improvement can be achieved with hard contact lenses in comparison to glasses. PMID- 9340927 TI - [Disposable contact lenses and their complications]. AB - Disposable soft contact lenses (DSCLs) have been marketed as a safer alternative to conventional soft lenses. Extended-wear DSCLs are designed for one or two weeks of continuous use before disposal. Those for daily wear are designed for use as conventional daily wear soft lenses, with daily removal and storage for 2 to 4 weeks before disposal. Beside minor complications, such as corneal abrasion, giant papillary conjunctivitis and toxic epithelial reactions to contact lens solutions, the most serious complication occurring in contact lens users is ulcerative keratitis. Several case-control studies performed over the last years, demonstrated that disposable contact lenses were associated with a 14-fold excess risk of ulcerative keratitis compared with that for patients wearing conventional daily-wear soft contact lenses and a 13-fold excess risk compared with that for wearers of rigid gas permeable contact lenses. However, the major risk factor for corneal ulceration in contact lens wearers is overnight lens wear of 1 to 3 nights. It was estimated that 49 to 74% of cases of contact lens associated ulcerative keratitis could be prevented by eliminating overnight wear. PMID- 9340928 TI - [Multifocal intraocular lenses as an alternative in cataract surgery]. AB - The development and introduction of multifocal intraocular lenses (MIOL) should provide the patient with a pseudoaccomodation for distance and near acuity without spectacle corrections. For this study two different types of multifocal lenses have been investigated concerning their visual properties. For diffractive MIOL (3M, type 815LE) an uncorrected distance visual acuity of Snellen 0.59 +/- 0.17 was found. The near visual acuity was J 2.4 +/- 0.94 and Snellen Acuity of 0.5 was achieved in a range of defocus of -1.25 D to +4.0 D. For refractive MIOL (Allergan, type Array SSM 26NB) an uncorrected distance visual acuity of Snellen 0.79 +/- 0.17 was found. The near visual acuity was J 2.75 +/- 1.35 and Snellen Acuity of 0.5 was achieved in a range of defocus of -1.0 D to +3.5 D. With this multifocal lenses the patients reached functional results for distance vision as well as with monofocals. Because of the existing pseudo-accommodation the results for near vision lay over of those of monofocals and the need spectacle correction decrease. PMID- 9340929 TI - [Refractive laser surgery of the cornea]. AB - The importance of refractive corneal laser surgery is increasing. Since the introduction of the Excimer laser in 1990 approximately 350.000 eyes were treated worldwide until the end of 1996. Postoperative refraction is mainly determined by the amount of intended correction: correction of myopia of up to -6 dpt using photorefractive keratectomy (PRK) has a success rate surpassing 93%, in higher myopic corrections the success rate drops to 30%. The complication rate is also directly related to the amount of correction: with corrections of up to 6 dpt it reaches up to 3% and in corrections of over 9 dpt manifest scars occur in more than 10%. Better results in myopias over -7 dpt are achieved using the laser in situ keratomileusis (LASIK), which is a combination of lamellar corneal surgery and the excimer laser. With LASIK the complication rate in corrections of up to 6 dpt is higher compared to PRK. The correction of hyperopia has still a fairly low success rate with both techniques that we suppose hyperopia to be a relative contraindication for corneal laser surgery. PMID- 9340930 TI - [The intrastromal corneal ring (KeraVision Ring, ICR, ICRS). A modern method for correcting minor myopia]. AB - For the correction of low myopia several competing surgical methods have been developed over the last two decades: radial keratotomy (RK) and photorefractive keratectomy (PRK) are used clinically on a very large scale. Both techniques are basically irreversible. RK does structurally weaken the cornea and PRK has to be applied directly at the optical center. Both disadvantages can to a large extent be avoided by the recently introduced Intrastromal Corneal Ring (ICR) that is now implanted worldwide following a long experimental and clinical development phase. The device is manufactured from PMMA and is effective by volume addition in the corneal periphery, shortening of the central arc length and thereby flattening the central optical zone. The latest development uses two ICR-Segments (with an arc length of 150 degrees each) that are placed in the deep stromal layers via a small radial incision. The surgical details, refractive results and intra- and postoperative complications of both, several international studies (with up to 5 years of follow-up), and our own experiences (25 eyes, follow-up 9 months) are presented. The ICRS (KeraVision Ring) seems to offer a precise and stable method to correct low myopia. The procedure is reversible to a large extent, potentially adjustable within certain limits and carries a minimal risk only. A short comparison with RK and PRK is attempted. Although long-term follow-up is not available yet, initial results of the technique seem very promising. PMID- 9340931 TI - [Therapy of Crohn diseases--results of a Consensus Conference of the German Society of Digestive and Metabolic Diseases]. PMID- 9340932 TI - [Abdominal arteriovenous and arterio-portal fistulas: etiology, diagnosis, therapeutic possibilities]. AB - PURPOSE: Arterioportal and arteriovenous fistulas (APF, AVF) are rare vascular disorders occurring as a result of congenital vascular malformation, trauma, iatrogenic causes or neoplasms. The clinical spectrum of presentation ranges from symptom-free individuals to patients with severe portal hypertension or congestive heart failure. While the majority of patients have been treated surgically in the past, interventional radiological procedures are being performed with increasing frequency. To prove this trend we reviewed the international literature. REVIEW: We reviewed the clinical presentation and management of 79 cases reported between 1980 and 1996 in the literature and six own patients. RESULTS: Review of the literature and our own six cases show that these fistulas can be divided in intrahepatic (n = 49, 75%) and extrahepatic fistulas (n = 20, 25%). The most important causes are trauma (n = 29, 37%), iatrogenic induced by procedures (n = 21, 27%) and congenital vascular malformations (n = 10, 13%). APFs and AVFs can be diagnosed by ultrasound, computed tomography and magnetic resonance imaging. Angiography confirms the diagnosis and in many cases allows definitive interventional radiologic treatment. CONCLUSION: Management of AVFs and APFs remains a challenge. However, interventional radiologic procedures provide a safe, low-cost, and effective method for treatment. Due to these facts in the last three years we notice an increasing part of interventional procedures for treatment. PMID- 9340934 TI - [Tumor vascularity as a prognostic factor and metastasis marker in stomach carcinoma]. PMID- 9340933 TI - [Preventive vaccination for viral hepatitis]. AB - Viral hepatitis A-E belong to the most important infectious diseases worldwide. Viral hepatitis is highly endemic in most developing countries in Africa, South East Asia, and southern America; however also in industrialized countries as Germany hepatitis A, B and C represent a thread which should not be underestimated. In Germany, there are about 20,000 to 40,000 hepatitis A infections every year, most of them acquired abroad; about 50,000 new hepatitis B infections and about 5,000 to 8,000 infections with hepatitis C virus occur every year. About 500,000 individuals are chronic carriers of hepatitis B virus and roughly the same number is supposed to be chronically infected with hepatitis C virus. As possibilities for therapeutic intervention in chronic hepatitis B and C are still limited, immunoprophylactic measures are of particular importance. Passive and active immunization is available for hepatitis A and B but so far not for hepatitis C. Passive immunization by application of specific immunoglobulins gives protection which is effective within a few hours but is limited according to the amount of immunoglobulin to six to twelve months. Active immunization on the other hand induces a specific immune response starting after a delay of usually days or sometimes weeks but nevertheless lasting for at least several years. The combination of both methods, passive-active immunization, has the advantage of immediate protection due to the immunoglobulin which lasts until the active immunization induces an endogenous antibody production. PMID- 9340935 TI - [Does fecal occult blood screening result in decreased mortality from large intestine cancer?]. PMID- 9340936 TI - [Barrett syndrome and risk of carcinoma. To screen or not to screen--that is the question]. PMID- 9340937 TI - [To pace or not to pace. Biatrial stimulation--a therapeutic option in atrial fibrillation?]. PMID- 9340938 TI - [Measuring stress hemodynamics in pulmonary circulation during the inpatient phase after myocardial infarct: a rightfully "forgotten" evaluation?]. AB - In 123 consecutive patients pulmonary hemodynamics at rest and during exercise were measured 7-10 days after acute myocardial infarction and systemic thrombolysis. Right- and left-heart catheterizations were performed at the same session and repeated after 6 months. We investigated if right-heart catheterization could predict the number of stenotic or occluded coronaries, the angiographic ventricular function and/or, most important, if the patient would profit from revascularization by PTCA or coronary surgery. RESULTS: 1) Pulmonary hemodynamics at rest and during exercise did not correlate to the number of stenotic or occluded coronaries, but exercise tolerance did. 2) Correlation between ventricular function and pulmonary hemodynamics was weak. 3) Only patients with pathologically elevated pulmonary pressures at exercise showed profit from revascularization (increase of ejection fraction > 5% at control). This profit was closely correlated to the angiographic extent of ventricular damage. Consequences from our results: Individual exercise tolerance can easily be measured non-invasively. Ventricular function can be measured non-invasively as well by echocardiography. Measurement of pulmonary hemodynamics at rest and during exercise does not contribute to planning the future management of the postinfarction patient and does not replace coronarography when indicated by clinical or non-invasive parameters. PMID- 9340939 TI - [Cardiovascular abnormalities in Bavaria 1984-1991]. AB - The study presents data on cardiovascular malformations in Bavarian livebirths, born between 1984 and 1991. Cases have been ascertained retrospectively by reviewing hospital records of all children being referred to a children's hospital up to 2 years of age. The classification scheme was based on abnormalities in developmental mechanisms. Among 984,570 livebirths, 7020 cases with structural congenital heart disease were identified. The birth prevalence was 7.1 per 1000 livebirths. Between 1984 and 1991, total prevalence increased from 5.9/10(3) to 8.0/10(3). Prevalence in males was 7.3/10(3) and in females 6.9/10(3). 78.1% of all heart defects were isolated, the remaining 21.9% were associated either with chromosomal abnormalities (9.6%), non-chromosomal syndromes (1.0%), or noncardiac malformations of other organ systems (11.3%). Total fatality rate was 12.0%, with two thirds of deaths occurring within a month of birth or the following month of life. Data were compared with those of the Baltimore-Washington Infant Study. This study presents for the first time regional data on birth prevalences of congenital heart defects in Germany. The classification scheme reduces the wide spectrum of phenotype cardiovascular defects to several pathogenetic groups. The defects in each group may be related to similar causal factors. PMID- 9340940 TI - [Anatomic and functional hypoplastic left heart syndrome and its surgical Norwood and Fontan treatment]. AB - The surgical therapy of newborns with hypoplastic left heart syndrome (HLHS) is still regarded with some distrust. The complete heart conserving palliation includes not only the Norwood operation during the newborn period but also the complete separation of both circuits by the Fontan operation some time later. Our experiences with each surgical step are presented. From 1989 to 1996, 43 infants with anatomical (n = 33) or functional (n = 10) HLHS underwent the Norwood operation. Functional HLHS were: Mitralatresia with double outlet right ventricle and subaortic stenosis (n = 2), atrioventricular septal defect with hypoplastic left ventricle, subaortic stenosis, and aortic coarctation (n = 1), hypoplastic, subaortic right ventricle with restrictive ventricular septal defect and aortic hypoplasia (n = 7). The median age at operation was 15 days (5 to 182 days), mean weight was 3.3 kg (3.0 to 4.9 kg). Total operative mortality was 32% (n = 14) with 16% since 1994 (3/19 patients). Five infants (12%) died 2 weeks to 6 months later, and 2 patients underwent cardiac transplantation. Up to now, 19 out of the 22 long term survivors underwent the bidirectional cavopulmonary anastomosis (Hemi-Fontan) at a median age of 7 months (2 to 14 months). Two infants died (10%). Up to now, 12 out of the remaining 17 survivors received the total cavopulmonary anastomosis after a mean period of 12 months. All children survived, and they are now completely palliated. The longest follow up after the complete Fontan operation is 6 years. CONCLUSION: With increasing experience the results of the Norwood operation improved. The following two-stage Fontan procedure bears only a low risk and leads to good quality of life. PMID- 9340941 TI - [Closure of the persistent ductus with detachable coils]. AB - 34 patients aged 3 months to 20 years underwent transcatheter-occlusion of their patent ductus arteriosus with detachable coils (Cook). Eight patients had a residual ductus after previous implantation of a Rashkind-occluder; 7 patients had various other cardiac malformations in addition. Only 6 patients had a large ductus with a diameter between 3 and 4 mm; all had systolic-diastolic murmurs. All other patients had ductus-diameters below 3 mm; three of them had systolic diastolic murmurs, 17 had systolic murmurs, and 8 patients had no murmur at all. The ductus was closed in 24 patients using arterial access only, in 6 patients via a venous, and in 4 patients both via venous and arterial catheterization. One coil was used in 23 patients, 2 coils in 9, and 3 coils in 2 patients. There were no complications of the intervention. Within 24 hours 31 patients (93%) had complete closure of the ductus and 32 patients (94%) after 6 months. Coil embolisation of the persistent ductus is a quick, safe and cheap method to close a ductus and has clear advantages compared to an operation. PMID- 9340942 TI - [Diagnostic value of auto-capture: detection of lead dislocation]. AB - Autocapture is a newly developed automatic threshold tracking algorithm implemented in implantable pacemakers. Based on the detection of the cardiac evoked response, autocapture continuously controls the effectiveness of stimulation. Increases in threshold are immediately followed by an adaption of the pulse amplitude. Automatic threshold tests periodically lower the amplitude in order to adapt to decreased thresholds. Thus, changes in threshold can be monitored over a long period and utilized as a diagnostic tool. This case report demonstrates the diagnosis and monitoring of a microdislodgement by means of autocapture. PMID- 9340943 TI - [Biatrial stimulation in therapy of paroxysmal atrial tachycardia: a case report]. AB - A biatrial pacemaker was inserted in a 68-year-old female with paroxysmal atrial fibrillation and atrial flutter refractory to antiarrhythmic drugs based on a sick-sinus-syndrome and concomitant interatrial conduction delay. One electrode positioned in the right atrium and another electrode located in the coronary sinus were connected to a dual-chamber pacemaker. The electrode in the right atrium was connected to the atrial channel, the electrode in the coronary sinus to the ventricular channel. The pacemaker was programmed in DDD-mode with an AV delay of 30 ms. Under a chronic antiarrhythmic medication with 160 mg sotalol per day there was no evidence for recurrent episodes of atrial tachyarrhythmias in the patient's history, 24-h-Holter-ECG, nor in the memory of the pacemaker during a follow-up of 8 months. PMID- 9340944 TI - [Modification of the detection and stimulation behavior of active and passive fixed bipolar pacemaker electrodes by depot dexamethasone]. AB - In a prospective, non-randomised study, two atrial steroid-eluting screw-in leads were evaluated (CapSureFix 4068, Medtronic, n = 17; Accufix II DEC 033-812, Telectronics, n = 16); in the same way, measurements were obtained of three steroid eluting ventricular electrodes (two screw-in leads: CapSureFix 4068, Medtronic, n = 11; Accufix II DEC 033-212, Telectronics, n = 20 and one tined lead Encor DEC 033-448, Telectronics, n = 18). Measurements were performed during implantation (= acute), 7 +/- 3 days after implantation (= subacute) and during follow-up at 3 and 6 months. After atrial implantation, there were no significant differences of the stimulation thresholds (Accufix II DEC: 0.76 +/- 0.23 V CapSureFix: 0.75 +/- 0.16 V). During follow-up, a slight but not significant increase of the stimulation threshold was observed (Accufix II DEC: subacute 0.99 +/- 0.45 V; 3 months 0.79 +/- 0.43 V; 6 months: 0.84 +/- 0.45 V.-CapSureFix: subacute 0.76 +/- 0.18 V; 3 months 0.87 +/- 0.31 V; 6 months 0.88 +/- 0.32 V). The acute atrial thresholds were significantly lower in the right atrial appendage than in the right lateral wall, while there was no difference during measurements at 3 and 6 months (atrial appendage n = 20 acute 0.69 +/- 0.11 V; right lateral wall n = 13 acute 0.85 +/- 0.25 V). The ventricular Encor DEC and Accufix II DEC electrodes had similar thresholds at implantation, whereas the CapSureFix electrode showed significantly higher stimulation thresholds (Encor DEC 0.56 +/- 0.15 V; Accufix II DEC 0.53 +/- 0.13 V; CapSureFix 0.8 +/- 0.16 V). During follow-up the stimulation threshold increased significantly with each ventricular electrode (Encor DEC: subacute 0.71 +/- 0.22 V; 3 months 0.77 +/- 0.23 V; 6 months 0.8 +/- 0.28 V-Accufix II DEC: subacute 0.86 +/- 0.44 V; 3 months 0.8 +/- 0.25 V; 6 months 0.74 +/- 0.21 V-CapSureFix: subacute 0.92 +/- 0.17 V; 3 months 1.14 +/- 0.46 V; 6 months 1.12 +/- 0.32 V). With regard to the sensing of the intracardiac signals, no differences among the electrodes were detected at the atrial as well as the ventricular level. Subacutely all electrodes had a significant decrease of sensing level without changes after 3 and 6 months (atrium: CapSureFix acute 4.08 +/- 1.34 mV, subacute 3.09 +/- 0.88 mV, 3 months 2.91 +/- 1.02 mV, 6 months 3.0 +/- 1.22 mV; Accufix II DEC acute 4.34 +/- 1.49 mV, subacute 2.86 +/- 1.18 mV, 3 months 3.07 +/- 1.04 mV, 6 months 2.91 +/- 1.16 mV-ventricle: CapSureFix acute 11.55 +/- 4.5 mV, subacute 9.99 +/- 3.51 mV, 3 months 9.36 +/- 3.23 mV, 6 months 9.13 +/- 3.4 mV; Accufix II DEC acute 10.66 +/- 3.0 mV, subacute 7.49 +/- 4.04 mV, 3 months 7.25 +/- 3.64 mV, 6 months 7.52 +/- 4.1 mV; Encor DEC acute 11.65 +/- 3.9 mV, subacute 9.04 +/- 3.29 mV, 3 months 8.69 +/- 3.83 mV, 6 months 8.78 +/- 3.32 mV). The impedance of the Accufix II DEC electrode was significantly lower than the CapSureFix electrode at the time of implantation in the atrium. The ventricular electrodes with active fixation showed a decrease of impedance 7 days after implantation, which diminished during chronic follow-up. On the other hand, the Encor DEC electrode did not exhibit any change of impedance at the different times of determination. In summary, the dexamethasone depots prevented the rise of the stimulation threshold in all the atrial electrodes with active fixation. There remained a small increase of the stimulation threshold after ventricular implantation, which did not reach clinical significance. Thus, the energy saving output of 2.5 volt could be programmed in almost every patient. PMID- 9340945 TI - [The quadri-cusp aortic valve: a congenital defect, responsible for pathologic changes?]. AB - We report on a 61-year-old patient with considerable insufficiency of a quadricuspid aortic valve and coronary three vessel disease. The congenital quadricuspid aortic valve has become symptomatic due to the development of (post endocarditic) insufficiency only in the advanced stage of life. It was treated by replacement of the aortic valve and bypass myocardial revascularization. The physiopathology of quadricuspid aortic valve will be discussed. PMID- 9340946 TI - [Ball variance and fracture of a Smeloff-Cutter prosthesis 24 years after aortic valve replacement]. AB - This report documents a case of ball variance in a Smeloff-Cutter aortic prosthesis occurring 24 years after implantation. After episodes of embolic complications the patient died in acute shock. The silicone rubber ball showed several alterations including discoloration, grooving, cracking, swelling and subtotal fracture of the poppet. Terminal valvular malfunction was caused by complete thrombosis of the prosthesis. In most patients ball variance occurred during the first years after valve replacement; thus, the observed case is a very rare late complication of a ball-valve prosthesis. PMID- 9340947 TI - [Recurrent angina pectoris in stress caused by hemangioma of the heart]. AB - We present the case of a 73-year-old man with a 3-week history of recurrent exertional angina confirmed by exercise stress testing. The symptoms were caused by hemangioma situated between the pulmonary artery and the left auricle. The diagnosis was confirmed by ventriculography, coronary angiography, and CT scanning. The tumor was completely removed through a midline sternotomy without extracorporeal circulation. Histology revealed the diagnosis of a cavernous hemangioma with endothelial lined vascular channels of varying size. The patient made an uneventful recovery and remained asymptomatic during follow up. Cardiac hemangioma are rare tumors of the heart that may cause angina by interfering with the coronary blood flow. Surgery is curative for most patients. PMID- 9340948 TI - [Accessory mitral valve tissue as a rare cause of subaortic stenosis and valvular aortic insufficiency]. AB - An unusual case of subaortic stenosis and aortic regurgitation caused by accessory mitral valve tissue in a 10 year old boy is reported. Two-dimensional and Doppler echocardiography revealed the characteristic feature of a mobile, parachute-like mass in the left ventricular outflow tract pro-lapsing into the aortic valve during systole and, thus, producing a systolic pressure gradient of 70 mm Hg between the left ventricle and aorta and causing mild aortic regurgitation. The accessory valve tissue was completely excised via an aortotomy without injury to the normal mitral and aortic valves. Two dimensional echocardiography provides excellent morphological information about the relationship between the accessory mitral valve tissue and the mitral and aortic valves, respectively. Accurate preoperative evaluation by two-dimensional echocardiography facilitates the successful surgical management of this rare condition. PMID- 9340949 TI - Special conference issue. International Meeting on Nucleic Acid Vaccines for the Prevention of Infectious Diseases. Bethesda, Maryland, 5-7 February 5-7, 1996. PMID- 9340950 TI - Workshop on the control and standardization of nucleic acid vaccines. Bethesda, Maryland, 8 February 1996. PMID- 9340951 TI - [Infections and vasculitis]. AB - Vasculitides are rare diseases characterized by inflammation of blood vessels. According to diameter of the blood vessels involved in the inflammatory process, the clinical presentation and the histological appearance, different vasculitic syndromes may be distinguished. Primary vasculitides are of unknown origin while secondary vasculitides may be caused by drugs, malignancy or infection. Panarteriitis nodosa caused by chronic Hepatitis B and mixed cryoglobulinemia secondary to chronic Hepatitis C are classical examples of vasculitides triggered by infections. However, these are rare complications of chronic viral hepatitis. Patients infected by HIV frequently suffer from vasculitis, which may be caused by opportunistic infections and by defects in immune regulation. In numerous case reports, various other infectious particles have been reported to cause different forms of vasculitis, either by direct infection of endothelial cells or by induction of an immunologic process leading to blood vessel destruction. Immunologically mediated vasculitis secondary to infection may be due to a predisposing reactivity of the patient's immune system. After successful treatment of the infection, the vasculitis usually subsides. Therefore, all patients with vasculitis should be evaluated for underlying infection. PMID- 9340952 TI - [Combination therapy with remission-inducing drugs in chronic polyarthritis: 1996 update]. AB - Therapy of rheumatoid arthritis with a combination of several disease-modifying drugs aims to better control of the disease than achievable by monotherapy. Subsequent to a paper written two years ago, this publication reviews studies dealing with combination therapy issued mainly in 1995 and 1996. Most studies deal with MTX as one of the partners. Beneficial results were reported for the combination of methotrexate with antimalarials, cyclosporine or sulfasalazine. The triple combination of methotrexate with hydroxychloroquine and azathioprine is especially promising although the studies presented up to now are still insufficient for its final assessment, due to methodologic problems. Similarly, the value of the combination of sulfasalazine with injectable gold, of sulfasalazine with methotrexate and hydroxychloroquine, or of methotrexate with injectable gold is still uncertain. PMID- 9340953 TI - [Development and evaluation of a modified version of the Larsen method for evaluating roentgenologic changes in chronic polyarthritis]. AB - AIM OF THE STUDY: To develop and evaluate a modification of the Larsen scoring method aimed at a clear definition of the different grades and a better correlation of the numerical scale with the amount of joint destruction. DESCRIPTION OF THE METHOD: While the original method described by Larsen is based on a comparison with standard reference films the modification defines the different grades semiquantitatively by the amount of destruction of the joint surface: grade II is a destruction (erosion) of the joint surface of up to 25%, in grade III it is 26-50%, in grade IV 51-75% and in grade V over 75%. Grade I is characterized by soft tissue swelling and in addition subchondral osteoporosis and remains unchanged compared with the original method. 32 joints of the hands, wrists and forefeet are counted summing up to a total score of 0-160. Examples for different joints are given. EVALUATION OF THE METHOD: Standard ap-x-rays of hands, wrists and forefeet of 24 patients with early erosive rheumatoid arthritis at baseline (T0) and after 36 months (T1) were graded by two investigators (G.H. and R.R.). The difference of the scoring between both observers (inter-observer difference) was related to the difference between the two timepoints (T0 and T1) in the same patient (intra-patient-difference) by means of a hierarchical analysis of variance (ANOVA). The intra-patient variance (from T0 to T1) was 259.3, which is 8 times greater than the inter-observer-variance of 32.1. The square root of intra-patient-standard deviation (16.1) and inter-observer standard deviation (5.7) is 2.8. These data show that the progression between T0 and T1 is real. The method was easily applicable to patients in a 2-year-DMARD trial with x-rays at baseline after 6, 12 and 24 months, showing a slowing of radiologic progression after month 6 under treatment with parenteral gold and methotrexate. CONCLUSION: The modification of Larsen's scoring method is a reliable measure to assess radiologic progression in patients with RA. Possibilities for further improvement are discussed. PMID- 9340954 TI - [The "bull horn sign"--scintigraphic pattern in sternocostoclavicular hyperostosis and pustular arthro-osteitis]. AB - 27 patients with sternocostoclavicular hyperostosis (SCC) and/or pustulotic arthroosteitis (PAO) were examined with whole body scintigraphy, conventional radiography, and other imaging modalities, such as CT, MRI. 25 of 27 patients with SCCH showed a characteristic high bullhorn-like uptake of the sternocostoclavicular region with the manubrium sterni representing the skull and the inflamed sternocostoclavicular joints corresponding to the horns (= bullhorn sign). Scintigraphy revealed additional skeletal manifestations (spondylitis, sacroilitis, osteitis, periostitis) in 19 of the 27 patients with SCCH and/ or PAO. In combination with PPP or psoriasis pustulosa, the typical scintigraphic bullhorn pattern enables the diagnosis of PAO (19 patients) with high confidence. Patients with SCCH but without skin disease at the time of presentation (8 of 27 patients) may develop PPP later and, therefore, it is justified to classify them as incomplete PAO with high risk to develop other skeletal manifestations later in the course of the disease. PMID- 9340955 TI - [Comparative evaluation of a German version of the Health Assessment Questionnaire and the Hannover Functional Capacity Questionnaire]. AB - OBJECTIVE: To translate the Health Assessment Questionnaire Disability Index (HAQ) into a German version, to validate and to compare its properties with two different versions of the Hannover Functional Ability Questionnaire (HFAQ) in a German speaking population. METHODS: The test-retest reliability was tested by Pearson correlation in 32 outpatients of the Department of Rheumatology of the Medizinische Hochschule Hannover. For retesting, the questionnaire was mailed to them 1 week later. To validate the questionnaire it was administered to 110 inpatients in three different hospitals. All patients fulfilled the American College of Rheumatology 1987 revised criteria of rheumatoid arthritis (RA) or the Rome criteria of definitive inactive RA. The internal consistency was measured by Cronbach's coefficient alpha (CCA). To assess criterion validity we compared the HAQ and the two versions of the HFAQ with Keitel's test (KT) and the modified Steinbrocker classification (mSC). Construct validity was assessed by comparing these instruments with different clinical and laboratory variables. A multivariate analysis was used to identify the most important factors that are influencing the HAQ- and HFAQ-scores. RESULTS: Test-retest reliability of the HAQ was r = 0.94. CCA was 0.91 (HAQ), 0.90 (HFAQ-P) and 0.93 (HFAQ-PR). The KT Pearson correlation coefficients reached r = -0.73 (HAQ), r = +0.74 (HFAQ-P) and r = +0.71 (HFAQ-PR). The mSC correlated r = +0.75 (HAQ), r = -0.72 (HFAQ-P) and r = -0.70 (HFAQ-PR). The correlation coefficients of HAQ/HFAQ-P was r = -0.87 and of HAQ/HFAQ-PR r = -0.88. The correlations between other clinical and laboratory variables reached from r = +/-0.58 (pain/HAQ) to r = +/-0.11 (number of swollen joints/HFAQ-PR). In backward multiple regression analysis 59-64% of the variance of disability measured by the questionnaires was explained predominantly by pain (32-33%) and by range of motion (16-21%). CONCLUSION: The German version of the HAQ presented here and the two versions of the HFAQ are reliable and valid instruments for measuring functional disability in a German-speaking population with RA. The construct measured by the HAQ and both versions of the HFAQ showed a high degree of correspondence. PMID- 9340956 TI - [Hemochromatosis arthropathy--an early manifestation of genetic hemochromatosis]. AB - Recent studies have shown a high frequency of genetic hemochromatosis in the Caucasian population. In addition, the well known organ involvement of genetic hemochromatosis was evident; more than 50% of patients develop a typical arthropathy which may result in severe physical disability. Among approximately 5000 patients referred to the rheumatology outpatient clinics of Bad Nauheim and Frankfurt with different rheumatologic diagnoses, 11 patients with typical signs of hemochromatotic arthropathy were identified. In none of those cases had the diagnosis "genetic hemochromatosis" been previously established. These patients had been treated for rheumatoid arthritis and other rheumatologic disorders over several years. All showed severe organ dysfunction due to iron overload, resulting in a reduced life expectancy. This investigation shows that knowledge of the typical signs of hemochromatotic arthropathy could lead to an earlier diagnosis of genetic hemochromatosis which is necessary to prevent the complications of iron overload in those patients. PMID- 9340957 TI - [Sprain of the cervical spine: early functional vs. immobilization treatment]. AB - Neck sprains are very common injuries often treated with immobilisation of different duration. The treatment with collars was tested against physiotherapy in a prospective randomised trial. Endpoints were defined as state of health, pain and costs. Ninety-seven patients with whiplash injuries were splitted by randomisation into two groups. One group was treated with a certain scheme of physiotherapy. Another group was treated with collar immobilisation for 3 weeks. Concerning symptoms at the time of admittance, age and sex distribution the groups were comparable. Fifty healthy persons with the same age and sex distribution served as a control group. Regarding to physical state of health and pain, which were examined by valid questionnaires, significant advantages of physiotherapy after two weeks were found. After 12 weeks the physical state of health corresponds to that of the control group. No influence on psychical state of health was seen. Physiotherapy for treatment of neck sprain is highly recommended. It has clear advantages over the treatment with collars with regard to state of health and pain, and it seems to be economically favourable. PMID- 9340958 TI - [Experience of the intensive care physician and patient outcome]. AB - Critically ill patients usually exhibit a wide variety of dynamic changes in various organ systems. Several outcome studies show that the presence of a trained, full-time intensivist is associated with a reduction in patient mortality. His experience leads to a decrease in otherwise frequently occurring iatrogenic complications. It thus seems beneficial to staff the intensive care unit with trained intensivists. PMID- 9340959 TI - [Secure anastomoses of the large intestine (especially with transanal drainage]. AB - The results of colonic resections were investigated in 356 patients in a retrospective analysis. Special attention was directed to the effectiveness of a transrectal tube, leading to decompression of the anastomosis for the first five days. Under elective conditions, the overall complication-rate was 12.7%. In emergency cases, these complications were 51.5% (one third of these cases had a decompression-tube). The clinical relevant leakage-rate under elective conditions ranged to 1.7%. Postoperative mortality related to surgical complications turned out to be 1.5% under elective conditions. The emergency operations had a high mortality which ranged to approximately 15.2%. No patient with leakage of a colorectal anastomosis died when the transrectal decompression-tube was applied. Such a safety in anastomotic healing of colon and rectum anastomoses can otherwise only be achieved by using the protection of a diverting colostomy or ileostomy. The use of the transrectal decompression-tube also avoids stomal complications and the second operation. There is no indication for the decompression-tube in emergency operations with purulent or faecal peritonitis. In these cases a fundamentally different treatment is mandatory. PMID- 9340960 TI - [The role of diagnostic laparoscopy in treatment of penetrating injuries of the lower thoracic wall and anterior abdominal wall]. AB - Penetrating injuries of the lower thoracic wall and anterior abdominal wall cause difficulties in the decision for laparotomy. For gunshot wounds laparotomy without further investigations is in most cases justified, but in other penetrating traumata one should use every diagnostic modality to prevent unacceptably high negative laparotomy rates. We performed diagnostic laparoscopy (DL) on 39 patients with penetrating injuries of the anterior abdominal wall and/or lower thoracic wall. Of these 39 patients, 25 had negative and 14 positive results. We had only one false-negative finding. No false-positive result occurred. We think that DL is a very reliable diagnostic tool which requires a relatively high technology. PMID- 9340961 TI - [Acute mesenteric ischemia]. AB - Acute mesenteric ischemia is a life-threatening vascular emergency. A retrospective analysis of our patients was performed to describe the development of the various procedures of diagnostic assessment and treatment between 1970 and 1996, to show the influence on survival and to define recent standards. PATIENTS: Between 1970 and 1996, 145 patients, 75 male and 70 female, suffering from acute mesenteric ischemia, have been treated at the Department of Surgery-University Hospital Vienna. RESULTS: In most cases AMI was caused by arterial embolism (64.1%, n = 93) followed by arterial thrombosis (27.6%, n= 40). Venous thrombosis (3.5%, n = 5) and non-occlusive AMI (4.8%, n = 7) were rare events. Serum lactate level has been determined routinely in all patients having been admitted with acute abdomen since 1984 and turned out to be positive in 81.2% (mean value 9.81 (3.21-22.3) mmol/l). Abdominal x-ray gave only in some individual cases special hints to the advanced intestinal gangrene. Abdominal sonography led to the correct diagnostic assessment in 52 patients (= 35.8%). Angiography was in 92% conclusive for the diagnosis. Abdominal CT led to establish the correct diagnosis in > 80%. Our series with revascularisation (thrombectomy/embolectomy or bypass) has resulted in 73.8% patient survival with intestine having been maintained in the most favourable cases. CONCLUSIONS: Early diagnostic assessment and treatment are decisive for survival. Abdominal-CT, angiography and serum-lactate constitute quick and reliable means to provide diagnosis and to judge the stage of AMI in addition to meticulous examination of patients' history, symptoms and physical conditions. PMID- 9340962 TI - [The plug and patch repair for managing the inguinal hernia of the adult]. AB - The basic principles of the Plug and Patch Repair of inguinal hernia are positioning of a prolene plug behind the internal ring in combination with a prolene onlay patch on the fascia transversalis. From July 1994 until December 1996 243 inguinal hernias were operated in this technique in a consecutive series of 221 patients. There were 211 primary and 32 recurrent hernias. 187 operations were performed in local anesthesia and 34 patients wished general anesthesia. No intraoperative complications were noted. The average operation time was 36 +/- 14 minutes. Postoperative complications were 8 subcutaneous hematomas and 1 superficial wound infection, which healed spontaneously. The average stay in hospital was 2 +/- 2 days and the patients returned to work after 17 +/- 11 days. Patients' self assessment of postoperative pain and limitations of daily activities revealed an excellent comfort. The follow-up rate is 97% and no late complications were observed after a median follow-up time of 21 months. The Plug and Patch technique is a very promising procedure for inguinal hernia repair in adults. PMID- 9340963 TI - [Physiologic and pathologic patterns of reaction to silicone breast implants]. AB - Local morphological reaction patterns on breast implants can be of high significance as possible starting point for controversely discussed systemic immune response triggered by silicon or silicone. Therefore, the collagenous capsules of 149 explanted mammoplasty prostheses were examined macroscopically, under a scanning electron microscope and light-microscopically using antibodies to the macrophage antigen CD68, vimentin, muscle actin, and the proliferation antigen MIB1, and were then correlated with anamnestic data (implanted type of prosthesis, indication for im- or explantation). According to our examinations, the in-vivo durability of the prostheses' shells is considerably decreasing with the expansion of their surfaces. Regardless of the type of the prostheses' surface regularly a chronic-proliferating inflammation pattern could be identified in the periprosthetic capsulectomy specimens starting with a synovial metaplasia of proliferating CD-68-negative and vimentin-positive mesenchymal cells in the area surrounding the implants and ending by its transformation into a stage of dense hyaline collagenous fibrous tissue after an advanced implantation period (> 2 years). By this, the texturing of the prosthesis surface modifies only the course, but not the quality of the chronically fibrosing inflammation. Bleeding of prosthesis as well as the incorporation of the polyurethane-foam coating of different prosthesis types into the periprosthetic breast capsule lead to a significant lymphoplasmacytic infiltration, partly with participation of local vessels as defined in a "silicone vasculitis". Morphological signs of an at least local immune response are detectable in 8.3% of the examined fibrotic capsules even without a morphologically identifiable foreign-body embedding. They can be possibly referred to- as well as the complete absence of hyaline collagenous fibrous tissue in 30% of the cases-a yet not causally clarified, inter-individually different susceptibility of the implant bearers. Only the systematic registration of the above-mentioned morphological reaction patterns in a "prosthesis-passport" together with the additional clinical observation of the patients can ensure in future the realistic estimation of potential health risks caused by silicone breast implants. PMID- 9340964 TI - [Aorto-duodenal fistula--direct suture and pedicled omentum flap-plasty]. AB - Between 1985 and 1993, six patients underwent emergency operation at the Department of General Surgery, Essen University Hospitals, for a secondary aorto duodenal fistula. In all patients, a Dacron tube- or bifurcation prosthesis has been implanted 1-10 years previously to repair an arteriosclerotic aneurysm of the abdominal aorta. The main symptom of the aorto-duodenal fistula was massive gastrointestinal hemorrhage, with manifest shock in two cases. The most reliable diagnostic procedure, in addition to ultrasonography, was found to be computed angio-tomography. In three cases where there was erosion around the proximal aorto-prosthetic anastomosis, bacterial contamination was found during surgery. Direct reconstruction and pedunculated omentum plasty appear to be a safe method for closing an aorto-duodenal fistula. No patient died in the immediate postoperative period after direct reconstruction. One patient, however, died three months after surgery of myocardial infarction. Two patients suffered from fistula recurrence 1.25 and 3 years respectively after operation and died as a result. One patient died of the sequelae of a chronic obstructive pulmonary disease 3.5 years after the operation. The remaining two patients are still alive and free of complications more than 4 years after operation. Direct reconstruction and pedunculated omentum plasty appear to be a safe method for closing an aorto-duodenal fistula. PMID- 9340965 TI - [Laparoscopy-assisted colorectal surgery. Early outcome in benign and malignant diseases--a prospective study of 120 patients]. AB - We present our results with laparoscopic-assisted colorectal surgery in 120 patients during the time from January 1993 to September 1996. The types of procedures cover almost the whole spectrum of colorectal surgery. They included hemicolectomies, signmoid resections, low anterior resections, Hartmann closures, proctocolectomies and rectopexies. 127 patients were subjected to laparoscopic operation, 7 needed conversion to open surgery (7% conversion rate). Average operation time was 145 +/- 58 min, length of postoperative stay 12 +/- 4 days. Oral food intake was started at the second day postoperatively without major problems. We observed perioperative complications in 21 cases (17%). There were 5 anastomotic leaks, 4 wound infections, 1 pneumothorax and 2 postoperative bleedings, 4 patients had clinical signs of prolonged bowel paralysis, 2 patients died as a consequence of anastomotic leaks, 2 of other reasons. In the postoperative period we saw a marked faster recovery and a lower complication rate compared to our conventionally operated patients and postoperative pain was less. In cases of malignant disease no rise in rate of recurrence was observed during follow-up (average 18 months) compared to open surgery. Until the results of long-term studies are published laparoscopic procedures should still be restricted to early tumor stages and palliative procedures. PMID- 9340966 TI - [Laparoscopic rectopexy]. AB - Within 4 years 66 laparoscopic rectopexies were performed. The indications were: rectal prolapse, morphologic outlet-constipation and a combination of both. Using a modified suture rectopexy (according to Sudeck), we did not take any foreign material and resected the sigmoid in 35 patients. Conversion rate was 2%, complications that needed reoperation occurred in 9%. In the follow up period of 24.1 months in the mean (max. 50) no recurrent prolapse occurred. Incontinence was abolished or improved in 64%, outlet-constipation was improved in 85%. Especially in rectopexy the laparoscopic technique seems to be of benefit for the patient: quicker convalescence, less pain, small scars, a.o. But all these potential advantages have to be proven in prospective-if possible randomised studies. PMID- 9340967 TI - [Thoracoscopic truncal vagotomy]. AB - Abdominal re-operation in patients with recurrent peptic ulcer disease is associated with a high morbidity rate and a mortality rate of 5%. As an alternative procedure, therefore, transthoracic truncal vagotomy was early recommended as a less invasive intervention, and good results can be achieved with it. With the development of minimal invasive surgery, this procedure can now be performed via thoracoscopy and patient stress thus reduced even further. Via a left-sided thoracoscopy, the parietal pleura is incised and a 3-5 cm long segment of the distal oesophagus mobilised and dissected free. Both the posterior and anterior trunks of the vagus nerves are identified and, after applying clips, transected. In order to achieve complete vagotomy, further fine branches have to be searched out and, if found, also divided. PMID- 9340968 TI - [Transmission of methicillin-resistant Staphylococcus aureus strains by foreign patients--measures for treating patients and preventing a nosocomial infection]. PMID- 9340969 TI - [Victor Schmieden (1874-1945) and his contribution to the development of modern surgery]. AB - Victor Schmieden (1874-1945) was the second Professor on ordinary for surgery (1919-1945) at the Frankfurt University, which was newly established in 1914. Among his numerous publications, especially the following contributed to the development of modern surgery: in 1915 during the first world war he proposed the operative treatment (laparotomy) of abdominal shot-wounds, in 1920 he established the operative treatment (pericardectomy) of patients with a calcified pericarditis (armour heart). In 1926 he described the adenomas (polyps) of the colon as a precursor of colorectal cancer (dysplasia-carcinoma sequence) and proposed total colectomy to patients with polyposis coli. PMID- 9340970 TI - [Recurrent and 2nd line therapy in ovarian carcinoma: an overview of conventional systemic therapy modalities]. AB - Remarkable improvements in primary surgery and chemotherapy for advanced ovarian cancer have been achieved in the last decades. Nevertheless, the majority of patients still develop recurrent disease and ultimately die from ovarian cancer. Evaluation of efficient second-line treatment is of clinical relevance. This review re-analyses the published data of conventional systemic treatment for recurrent and refractory ovarian cancer. Patients were grouped according to prior chemotherapy (with or without platinum; with or without paclitaxel) and according to clinical platinum resistance. Platinum resistance is defined as having progressed during platinum based chemotherapy or having relapsed within 6 months after completion of primary platinum containing treatment. The most favourable results in platinum sensitive ovarian cancer were reported for platinum containing re-induction chemotherapy. Results in platinum refractory ovarian cancer were different. Both hormonal treatment consisting of either tamoxifen or GnRH analogues, and chemotherapy with topoisomerase blocking agents, taxanes, or alkylating agents did show similar results. Unfortunately, there are only limited data from prospectively randomised trials in these patients. Therefore, recommendations remain inconclusive. Retrospective comparisons may help the clinician to chose the currently best available treatment for an individual patient, however, these treatments have to be evaluated in prospectively randomised trials. The protocols of the ongoing studies in refractory or recurrent ovarian cancer of the AGO Ovarian Cancer Study Group are outlined. PMID- 9340971 TI - [Gestational diabetes--perinatal hyperinsulinism and postnatal developmental disorders]. AB - The impaired glucose tolerance in pregnancy (IGT) represents an important fact in aetiopathogenesis of insulin dependent diabetes mellitus (IDDM) and non insulin dependent diabetes (NIDDM) as well as obesitas and cardiovascular diseases in context with fetal hyperinsulinism. Prospective studies of diabetic mothers newborns are difficult by reason of health controls in different outpatient departments. The aim of this review is to claim a general glucose screening in pregnancy looking on the development of newborns in later life. In present preventive prospects were not used to decrease the morbidity in diabetes, obesitas and cardiovascular diseases without gestational diabetes screening in pregnancy. The neonatal onset and late morbidity is dependent on the quality of maternal glycemia in pregnancy measured by means of glycosylated hemoglobin and insulin the amniotic fluid. PMID- 9340972 TI - [Value of laparoscopic pelvic lymph node excision in inguinal vulvar carcinoma metastasis]. AB - We performed a laparoscopic pelvic lymphadenectomy in three patients with vulvar cancer FIGO stage II or III and positive inguinal lymph nodes. Computertomographical and lymphographical findings of pelvic lymph nodes did not correlate with the histological findings in two patients: Surgical staging up graded the clinical stage in one patient. No intra- or postoperative complications were encountered. Laparoscopic pelvic lymphadenectomy in inguinal metastasizing vulvar cancer is a valuable method with low peri- and postoperative morbidity. PMID- 9340973 TI - [Rate of proliferation as a prognostic criterion in endometrial carcinoma--an immunohistochemical analysis with the monoclonal antibody KI-S1]. AB - In a subset of 183 primarily operative treated patients of 300 patients with endometrial carcinoma an analysis of histological tumor type (n = 142), myometrial invasion (n = 115), stage (n = 114), immunohistochemically detected steroid receptor status, histologic grading (n = 152) and growth fraction, detected by immunohistochemical determination of antibody Ki-S1 (n = 130), was performed. The mean follow up was 9.1 years. By univariate analysis, age, myometrial invasion (< = 3/>3 mm), expression of steroid receptor, FIGO-stage, histologic grading and growth fraction were found to influence the overall survival rate significantly. Cox multivariate regression showed age, FIGO-stage and growth fraction to be independent predictors of overall survival. With respect to survival univariate analysis revealed progesterone receptor status, FIGO-stage, histologic grading and growth fraction as prognostic factors. By multivariate analysis FIGO-stage, histologic grading and growth fraction were found to be independent prognostic factors for survival. Multivariate and univariate analysis exhibited FIGO-stage, histologic grading and growth fraction rate to be independent predictors of a disease-free survival. Immunohistochemically detected growth fraction seems to be useful as an additional prognostic factor in endometrial cancer. PMID- 9340974 TI - Invasive breast carcinoma with granulomatous stromal response. AB - An unusual case of invasive ductal carcinoma of the breast associated with epitheloid granulomas is reported. Multinucleated giant Langhans'-type giant cells were found in the epitheloid granulomas in breast carcinoma, but there were not present in the breast tissue and axillary lymph nodes. Congo-red deposits were found haphazardly in the stroma between tumor cells and granulomas. Numerous mast cells were found surrounding granulomas. The patient lacked any clinical evidence of the systemic granulomatous disease. The presence of epitheloid granulomas, amyloid deposits and numerous mast cells in invasive breast carcinoma could be related to a host immune response towards the tumor. PMID- 9340975 TI - [The historical development of radical vaginal operation of cervix carcinoma]. AB - Reviewing the historical development of radical vaginal surgery for the treatment of cervical cancer the different variations of this technique are defined. In 1880 the first modified radical hysterectomy for cervical cancer was performed by Pawlik. Radical vaginal hysterectomy is associated with the name of Schauta who further developed and categorized the technique. The most important modifications were done by Stoeckel and Peham/Amreich. Extraperitoneal dissection of pelvic lymph nodes was combined with radical vaginal hysterectomy by Stoeckel in 1928. PMID- 9340976 TI - [Effect of progestins on the breast--protective or proliferative?]. AB - This survey reviews the published data on progesterone and progestins in normal, benign and malignant breast disease. Controversial data will be put together and the results demonstrating proliferative action and a proliferative and mitotic action on one hand and an inhibiting and protective action on the other hand will be presented. PMID- 9340977 TI - [Hysteroscopic myoma resection of submucous myomas with largely intramural components]. AB - From February 1992 to December 1995 hysteroscopical myoma resections were performed in 70 patients suffering from recurrent bleeding disorders. In 24 cases of single myoma these were submucous with their largest portion located in the uterine wall. In all cases a pretreatment was performed with 2-3 injections of GnRH-analogues. The indication for resection must be proved critical in submucous myoma with their largest portion in the uterine wall, because a higher rate of complications is described. A simultaneous sonographical or laparoscopical control is necessary. In 2 cases of large myomas second resections were performed. The hysteroscopical resection of submucous myoma with their largest position in the uterine wall is a procedure for a surgeon with much experience in the field of operative hysteroscopy. In a follow up of 5 to 52 months a normal menstruation was reached in all patients. No intra- or postoperative complications were seen, no hysterectomy had to be performed until now. The resection of submucous myoma with their largest portion in the uterine wall is a surgery without a higher complication rate when carried out by an experienced surgeon. PMID- 9340978 TI - [Williams-Richardson vaginopexy. An abdominal suspension operation in prolapse and extensive vaginal descent]. AB - Vaginopexy according to Williams and Richardson is an abdominal colposuspension by strips from external oblique aponeurosis. It is indicated in cases of severe descent or prolapse of the vagina when the ability to have intercourse has to be maintained. At the Department of Gynecology and Obstetrics, AKH Celle, between 1989 and 1996 99 patients of whom 90 had already been operated before were treated by the Williams-Richardson method. An examination of 60 patients with a follow-up time period of 6 months to 6 years showed satisfying results in 54 patients with adequate vaginal support. In 12 cases postoperative hernia of the abdominal wall occurred. Since a vicryl mesh is used in addition to the external oblique aponeurosis, this complication has no longer been observed. PMID- 9340979 TI - [Doppler score for evaluating perinatal risk]. AB - Doppler sonography now has a definite place in the surveillance of risk pregnancies. Uniform clinical management is sometimes difficult especially in borderline cases. The following study demonstrates the possibility of standardizing and systematizing Doppler results using a score. In a collective of 253 pregnant women we performed Doppler examinations in the fetal aorta, umbilical artery, middle cerebral artery, internal carotid artery. The results were divided into 4 groups and correlated to the fetal outcome. There was a highly significant worsening in prognosis regarding duration of pregnancy, birth weight and rate of cesarean sections with increasing Doppler score. In the event of pathological and highly pathological scores, the average duration of pregnancy was 23 and 48 days shorter than normal. As a result, there was a highly significant reduction in the average birth weight compared to fetuses with normal Doppler scores: by 1060.7 g in the case of a pathological score and by 1633.5 g in the case of a highly pathological score. There was a highly significant correlation concerning the rate of cesarean sections and the indication "fetal distress". The average interval between diagnosis and birth was 6.3 days in the case of pathological Doppler findings and 2.3 days in the case of highly pathological findings. The difference was highly significant. In the case of highly pathological scores all fetuses were delivered after at least 5 days, compared with after at least more than 10 days in those with only pathological Doppler findings. This reflects the fact that there is none room for discretion in case of a highly pathological flow. In summary the Doppler score allows better estimation of fetal risk and can improve fetal prognosis by special monitoring and earlier obstetric intervention. PMID- 9340981 TI - [Gynecologic oncologic information from the Internet]. PMID- 9340980 TI - [Prolongation of a bi-amniotic twin pregnancy after premature rupture of fetal membranes and umbilical cord prolapse of the first twin during the 23rd week of pregnancy]. AB - A 41-year old woman had a premature rupture of the membranes of the first twin with prolapse of the umbilical cord and the left foot in the 24th gestational week (23 + 4). The treatment consisted of bed rest in the Trendelenburg position, antibiotic prophylaxis and glucocorticoids for lung maturation. After 7 days the first twin was delivered vaginally from breech position. He died 10 hours later due to intraventricular hemorrhage. Tocolysis was administered and the umbilical cord was ligated and cut as high as possible. Seven days later a premature rupture of the membranes of the second twin and a prolapse of the fetal hand occurred. The second twin (birth weight 750 g) was delivered by cesarean section in the 26th gestational week (25 + 4) and survived without neurologic sequelae. Prolongation of pregnancy after a very premature delivery of the first twin of a biamniotic twin pregnancy can improve the neonatal outcome. PMID- 9340982 TI - [The study group for Information Processing in Gynecology and Obstetrics--Section of the German Society of Gynecology and Obstetrics e. V. (AIG): Responsibilities and current projects]. AB - The German Study Group of Informatics in Obstetrics and Gynecology supports solutions of Computer applications in Obstetrics and Gynecology. Actual projects are evaluation of software, congress organisation and improvement of communication using the word wide web. PMID- 9340983 TI - [Standards in treatment and assessment of transsexual patients. German Society of Sexual Research, Academy of Sexual Medicine and Society of Sexual Science]. PMID- 9340984 TI - [Microbiological factors determining the carrier state in droplet infections]. PMID- 9340985 TI - [The suppressive action of a magnetic-laser ray and of an electrolytic solution of sodium hypochlorite on the factors of causative agent persistence]. AB - The suppressive action of a magnet-laser ray and electrolyzed sodium hypochlorite solution on the persistence factors (antilysozyme, "anti-interferon") of Staphylococcus aureus and Neisseria gonorrhoeae is shown. The optimum conditions (time, dose, concentration) for the regulation of the persistence properties of the pathogens has been determined. The use of physicochemical factors in the proposed parameters has been shown to be effective for the therapy of purulent inflammatory diseases and the sanitation of S.aureus carriers. PMID- 9340986 TI - [The immunological consequences of congenital infections]. AB - Mice of different strains were inoculated with type A influenza virus or Mycoplasma arthritidis in the second half of pregnancy. A part of the animals born after this inoculation were characterized by a sharp retardation of growth. The study of the immune status of such animals revealed that their proliferative response to mitogenic/superantigenic factors of the infective agents introduced during pregnancy was suppressed or absent, and the cells of their immune system began to recognize syngeneic intact stimulators in the mixed lymphocytes culture as heterogeneous ones. The spleen of the experimental animals was found to contain suppressor cells, both specific and nonspecific with respect to the infective agent. After inoculation with M. arthritidis areactivity was observed only in mice, sensitive to mycoplasmal superantigen. The data thus obtained suggest that the penetration of infecting agents producing mitogenic/superantigenic factors induced changes in the immune system, contributing to the persistence of the infective agent in the host body. PMID- 9340987 TI - [Microorganism persistence in hematopoietic tissue: means for detecting it and its significance]. AB - Many microorganisms are capable of prolonged persistence in the marrow. In this study, carried out by the method of negative selection based on the treatment of mouse marrow cells with specific antimicrobial sera and complement, Mycoplasma arthritidis and L-forms of group B streptococci were found to be capable of persisting in the marrow in close association with the late category of clonogenic precursor cells, CFU-7, as well as, to a lesser extent, with late erythroid precursors, CFUe. Early colony-forming cells, CFUs-12 and PFUe, as well as granulocyto-macrophagal precursors, CFUgm, did not practically express antigens to the given infective agents on their surface. PMID- 9340988 TI - [The differentiated correction of the local immunity of the reproductive tract by using lysozyme and immunoglobulins in pregnant women with a genital infection]. AB - Pregnant women with genital infection were locally treated with lysozyme and immunoglobulins. The use of these preparations was found to normalize the characteristics of local immunity, to influence the composition of the genital microflora and to lead to the disappearance of the clinical symptoms of the disease and to a decrease in the frequency of relapses of the inflammatory process. PMID- 9340989 TI - [A nutrient medium for the isolation of escherichiae with persistence properties]. AB - The possibility of establishing the etiological and epidemiological importance of Escherichia on the basis of the determination of their persistence factors made it necessary to develop simple and, at the same time, informative methods for their indication. For this purpose a new selective medium for the isolation of Escherichia with persistence properties was developed and approved. The use of this medium permits the early detection of persistent strains in the process of their isolation from a human body and environmental objects. The nutrient medium may be used in clinics, as well as in sanitary and hygienic practice, especially in the system of microbiological screening studies. PMID- 9340990 TI - [The photometric determination of the antilysozyme activity of microorganisms]. AB - The method for the determination of the antilysozyme activity (ALA) of microorganisms, based on the photometric determination of the residual activity of enzyme with the use of Micrococcus lysodeikticus test culture after the incubation of the strain under test and lysozyme, is proposed. The new method enhances the reliability of the determination of the ALA of microorganisms due to an increase in the accuracy of the quantitative determination of ALA. The elimination of the stage of interaction between the growing bacterial culture and lysozyme and the presence of the antilysozyme factor in the supernatant fluid confirm the constructive and secretory character of antilysozyme activity. PMID- 9340992 TI - [The interaction of Escherichia coli, possessing the antilysozyme trait, with infusoria]. AB - In this work isogenic E.coli strains K-12J53 and J53pAlz60, differing in the presence of the antilysozyme characteristic, were used. In the process of joint cultivation the degradation of lysozyme in protozoa by antilysozyme-active E.coli was established. The initial culture of antilysozyme-active E.coli K-12pAlz60 was found to be slightly heterogeneous with respect to the antilysozyme characteristic; this heterogeneity increased after the interaction of E.coli with protozoa. As the result of their joint cultivation with Tetrahymena, clones with low (2 micrograms/ml) antilysozyme activity (ALA) disappeared from the population and clones with medium and high values of ALA (3, 4, 5, 6 micrograms/ml) accumulated. The dynamics of the interaction of infusoria with Escherichia cells (ALA+) on the ultrastructural level in 1-6 and 24 hours revealed the gradual increase of processes leading to the destruction of most of the bacteria (up to their complete digestion) and, at the same time, the presence, observed also at an early period, of intact bacteria, resistant to lysis, whose survival was ensured by their antilysozyme characteristic. PMID- 9340991 TI - [The determination of the fresh fecal contamination of the water of surface reservoirs]. AB - The approbation of a method for the determination of new fecal contamination of water in surface reservoirs has been carried out; in accordance with this method, the presence of new fecal contamination in water is determined by the level of manifestation of the persistence properties (antilysozyme, anticomplementary "anti-interferon" activities) of Escherichia coli, used as a sanitary indicator microorganism. PMID- 9340993 TI - [Bacterial virulence as a function of adaptation]. PMID- 9340994 TI - [A general analysis of the concepts about pathogenic and opportunistic bacteria]. PMID- 9340995 TI - [The mechanisms of the stable preservation of the causative agent of plague in the environment (new facts and hypotheses)]. PMID- 9340996 TI - [The persistence of bacterial pathogens as a result of relationships in the parasite-host system]. PMID- 9340997 TI - [The problems of the adaptation of pathogenic bacteria to the environment]. PMID- 9340998 TI - [The nonculturable state in pathogenic bacteria modelled on Salmonella typhimurium: the phenomenon and its genetic control]. AB - The data on the influence of mutation in genes, affecting vital functions of a bacterial cell, on the process of transition into the nonculturable state in S. typhimurium are presented. Four genes obtained by the authors have been identified. Mutations in these genes lead disturbances in the formation of S. typhimurium nonculturable forms. The nonculturable forms of salmonellae have been shown to retain their virulent properties and their capacity for recultivation, when inoculated into laboratory animals. PMID- 9340999 TI - [The possibility of preserving the causative agent of plague in soil in resting (nonculturable) form]. AB - The parallel study of soil specimens from the burrows of great gerbils, taken in the natural focus of infection at the period between epidemics, by the bacteriological method and with the use of polymerase chain reaction (PCR) demonstrated that, simultaneously with the negative results of inoculations into nutrient media, PCR revealed the presence of Y. pestis in 4 out of 72 soil specimens. The serological study (in the indirect hemagglutination test) revealed the presence of Y. pestis capsular antigen (fraction l) in 16 out of 75 soil specimens. The transition of Y. pestis (strain EV) into the nonculturable state was experimentally shown in sterile soil extract and in association with protozoa, algae and Euglena. Bacteriologically detected Y. pestis disappeared or were represented by a few cells after day 7, while PCR could register the presence of Y. pestis at a concentration of 10(4)-10(5) microbial cells per ml until the end of the term of observation (30 days). After the enrichment of the nutrient medium with fetal serum the complete reversion of Y. pestis nonculturable forms was observed: the concentration of bacteria grown on agar was 10(5) colony-forming units per ml. Transition into the nonculturable state was also observed in distilled water. The results of this investigation are discussed with special emphasis to the possibility of the preservation of Y. pestis in the nonculturable from during the period between epidemics as applied to the problem of plague epizootic. PMID- 9341000 TI - [The persistence of mycoplasmas in the infected organism: observations, causes and mechanisms, diagnosis]. PMID- 9341001 TI - [The dynamics of the species composition, antilysozyme activity and antibiotic resistance of the causative agents of soft-tissue surgical infection]. AB - The dynamics of the microflora in the foci of surgical infection in 54 patients was studied. Staphylococci and streptococci prevailed among the primarily isolated causative agents. In complicated forms of surgical infection of soft tissues the change of causative agents, mainly to gram-negative opportunistic microflora with a higher level of resistance to antibiotics and high antilysozyme activity, was observed more often than in noncomplicated forms of such infection. PMID- 9341002 TI - [Genetic markers in the epidemiology of bacterial infections]. PMID- 9341003 TI - [The ecological determinacy of the markers of staphylococcal persistence]. AB - In the analysis of the presence and manifestation of the capacity for the inactivation of lysozyme, complement, the bactericidal component of interferon and immunoglobulins in 354 Staphylococcus aureus cultures and 175 S. epidermidis cultures the system of exogenous factors which determine the intraspecific dispersion of the properties under study has been characterized. The combined action of these factors leads to the selection of variants, most adequate to the conditions of a specific ecological niche, which is manifested both by the convergence of phenotypic characteristics in staphylococci of different species found in similar environment and by their divergence in the representatives of a certain species, isolated from different sources. PMID- 9341004 TI - [The anticomplement activity of Neisseria gonorrhoeae]. AB - Bacteria of the species N. gonorrhoeae have anticomplementary activity whose absolute values exceed the level of this activity both in other representatives of the genus Neisseria and in microorganisms of other taxonomic groups found in the microbiocenosis of the reproductive tract. The specificity of the anticomplementary action of N. gonorrhoeae extracellular products with respect to individual components of the complement system, as well as their role in the formation of the state of seroresistance and in the determination of the effectiveness of the interaction of gonococci with neutrophilic phagocytes in the system of opsonic cooperation, have been characterized. PMID- 9341005 TI - [An analysis of the interrelation of antilysozyme activity and reproductive function in escherichiae]. AB - In vitro experiments on 72 E. coli strains showed the presence of relationship between the level of their antilysozyme activity and a number of parameters of the reproduction of these bacteria. The factor analysis revealed the integration of the antilysozyme sign with the factors which determined the inhibition of the development of bacterial populations at early stages and controlled the accumulation of bacterial biomass. The study established that the beginning of the expression of the antilysozyme sign of E. coli was associated with the transition of the culture to the phase of slow growth, and the subsequent dynamics of the level of antilysozyme activity was only partially linked with the increase of biomass. The suggestion was made that the antilysozyme factor was one of the elements of the intracellular system regulating the synthesis (and/or stabilization) of peptidoglycan of bacterial cell walls. PMID- 9341006 TI - [The ribonuclease activity of bacteria as a factor in the persistence of the causative agents of sapronotic infections]. PMID- 9341007 TI - [The characteristics of the hydrophobic properties of bacteria when interacting with blood serum]. AB - Considerable variability of the hydrophobic properties of bacterial surface was demonstrated on 132 representatives of Escherichia coli, Neisseria, staphylococci and enterococci, belonging to museum strains and isolated from clinical material. Taxonomic and pathovariant differences in their susceptibility to hydrophobic modification induced by the thermostable and thermolabile components of blood serum were experimentally established. Extracellular bacterial products were found to be capable of preventive and restorative counteraction to such hydrophobic modifications. The antilysozyme and anticomplement activities of bacteria were shown to take part in the expression of hydrophobic/hydrophilic properties in the interaction of bacteria with blood serum. PMID- 9341008 TI - [The characteristics of an experimental infection in mice induced by salmonellae with different persistence features]. AB - The comparative analysis of changes in the immunoreactivity of the body in (CBA x C57BL6)F1 mice in experimental infection induced by strains, differing in their plasmid profile and persistence properties, was carried out. All infected mice were found to have, on the whole, similar changes in lymphoid organs. The most severe course and outcome of the infectious process, accompanied by pronounced shifts of immunological characteristics and the activation of peroxide oxidation of lipids, were observed in mice inoculated with Salmonella typhimurium strain 178, having a polyplasmid profile and the average level of persistence factors activities. PMID- 9341009 TI - [The role of Staphylococcus epidermidis persistence factors in the infectious process]. AB - The species composition of microflora was determined and the etiological role of S. epidermidis was shown in some purulent inflammatory diseases (maxillary sinusitis, urethritis). The role of the complex of S. epidermidis persistence factors in the chronization of the infectious process and the formation of the resident carrier state was established. PMID- 9341010 TI - [Factors in the persistence of opportunistic microorganisms in dysbacteriosis of the gastrointestinal tract]. AB - The presence of correlation between the quantitative content of individual carboxylic acids and aromatic compounds of microbial origin in feces and the degree of the antagonistic activity of feces was proved in experiments on conventional laboratory rats receiving therapeutic doses of tetracyclin and pefloxacin for 8 days. Antagonistic activity was regarded as one of the factors of the colonization resistance of the intestine to the invasion of opportunistic microorganisms, and microbial metabolites studied in this work were regarded as factors contributing to the persistence of these microorganisms. PMID- 9341011 TI - [The role of persisting opportunistic intestinal microflora in dysbiosis in the occurrence of hepatobiliary system diseases]. AB - The state of microbiocenosis in the intestine of 89 patients with surgical pathology of biliary ducts was determined. Dysbiotic disturbances of different severity were detected in 96.6% of the examined persons. High occurrence of dysbiosis of the large intestine and the presence of the capacity for inactivating natural host resistance factors in different representatives of intestinal microflora were regarded as the cause of the development and maintenance of the inflammatory processes of the biliary ducts. PMID- 9341012 TI - [The effect of therapeutic mud on the viability and persistence properties of bacteria]. AB - The influence of therapeutic salt mud on the viability and some biological properties of bacteria, responsible for their survival in the macroorganisms, was shown. Therapeutic mud had low bactericidal properties, and enterobacteria were, on the whole, even less sensitive to these properties than staphylococci. Therapeutic mud inhibited the capacity of bacteria for inactivating complement, lysozyme and the bactericidal component of the preparation of interferon and also reduced the hydrophobic properties of bacterial cells. At the same time Escherichia were found to be more susceptible to the modifying action of the mud than staphylococci. The greatest effect on the hydrophobic properties and anticomplement activity of bacteria was observed after their incubation in mud solution. PMID- 9341013 TI - [The effect of steroid hormone preparations on the persistence and growth characteristics of staphylococci]. AB - The influence of sex steroid hormones on the antilysozyme activity and growth characteristics of Staphylococcus aureus was studied. The study revealed that in the process of the cultivation of S. aureus in the presence of the hormones the antilysozyme of these bacteria became lower and the degree of heterogeneity of the population with regard to the sign under study decreased. The nonuniform influence of the hormones on the growth characteristic of staphylococci was registered. The data obtained in this study may be used for explanation of mechanisms of the formation of the microbiocenosis of the reproductive system. PMID- 9341014 TI - [The antilysozyme activity of the causative agent in the diagnosis and prognosis of the course of gonococcal infection]. AB - Differences in the frequency and level of antilysozyme activity in Neisseria gonorrhoeae cultures isolated from patients with different forms and stages of gonococcal infection were revealed. The possibility of using the antilysozyme sign of N. gonorrhoeae for the differential diagnostics of the clinical forms of this infection and for prognosticating the chronization of the inflammatory process in newly infected patients and in cases of relapses in chronic gonorrhea patients. PMID- 9341015 TI - [The role of the persistence characteristics of microflora in the etiological diagnosis and determination of the sources of the infection of the urinary system organs in pyelonephritis in infants in the first year of life]. AB - The microbiological status of 63 infants with pyelonephritis was determined. The importance of bacterial persistence factors for the identification of the infective agent and the detection of the source of infection of the urinary tracts was evaluated. The intestinal reservoir was shown to be the source of persistent microflora causing the formation of pyelonephritis, the change of the pathogenic species and the development of isolated bacteriuria at the end of the acute stage of pyelonephritis in children under 1 year of age. PMID- 9341016 TI - Transactions of the XXVI Congress of the Scandinavian Oto-Laryngological Society. Tampere, Finland, August 8-11, 1996. PMID- 9341017 TI - [The diverse aspects of angina pectoris]. PMID- 9341018 TI - [The physiopathology of angina pectoris]. AB - The author reviews the pathophysiology of angina pectoris. The first part concerns to the regulation of coronary blood flow: 1. Determinants of myocardial oxygen consumption (preload, afterload,contractility, heart rate); 2. Factors that control the myocardial oxygen supply (perfusion pressure, coronary vascular resistance). The second part concerns the coronary insufficiency and its clinical consequences (angina pectoris, myocardium infarction, heart failure, arrhythmias, primary cardiac arrest. The third part concerns the principal pathophysiologic mechanisms of the stable angina and of the unstable angina (progression of atherosclerosis, platelet aggregation, thrombosis and/or alterations in vasomotor tone). PMID- 9341019 TI - [Angina pectoris. The extent of the problem]. AB - Angina pectoris is analysed according to the principal data of its social repercussion: frequency, gravity, chronicity, diagnostic and treatment complexity, treatment costs and genetic factor extension. Some data of the national literature defining the dimension of the problem are given and a comparative study with other diseases and other countries is made. In conclusion, the need to develop epidemiological studies of adequate dimension is supported, mostly to better define frequency of the disease (prevalence and incidence) since the hospital and the mortality data are not enough. However, from the available data, it is expected that the prevalence of ischemic heart disease, of which the angina is a frequent clinical manifestation, will increase as well as the economic costs. In fact, the costs will be the consequence of scientific and technological development and of the increasing search for health by the population can services. PMID- 9341020 TI - [The stress electrocardiogram in the evaluation of angina pectoris]. AB - The exercise tolerance test (ETT) is the ancillary test of choice for the evaluation of the patient with ischaemic heart disease. When faced with complaints suggestive of angina pectoris, ETT is useful in confirming or excluding such a diagnosis, frequently reproducing the complaints while recording the tell-tale electrocardiographic signs of ischaemia--ST segment deviation. Additionally, the ETT provides information on the severity of the disease and is also valuable for the risk stratification of future coronary events. This information as a whole can be used both in the diagnosis and in the management of the patient with coronary artery disease. PMID- 9341021 TI - [Myocardial perfusion scintigraphy for diagnosis and risk stratification in stable angina pectoris]. AB - In this paper, the indications for myocardial perfusion scintigraphy, the stress methods, the radiopharmacy and the most used methodology are reviewed. The clinical applications of myocardial perfusion scintigraphy, regarding the diagnosis and prognostic stratification in stable angina are emphasised. PMID- 9341022 TI - [Coronary angiography in the outpatient with angina pectoris]. AB - Coronary angiography remains the only definitive diagnostic procedure to assess the severity and extension of coronary artery disease. We discuss some technical aspects and the place of angiography in the risk stratification of the coronary patient comparing conservative and invasive strategies. It is suggested that a progressively more important role will be played by the general practitioner regarding indication, interpretation and therapeutic decision, sharing responsibilities with the cardiologist and the patient. With increased facilities for coronary angiography and increased ad hoc angioplasties, observed at present, the GP should also be responsible for proper selection of the reference cardiologists and hospitals. PMID- 9341023 TI - [Angina pectoris. Silent ischemia]. AB - One of the most difficult problems related to coronary artery disease is the detection and eventual treatment of silent myocardial ischemia (SMI). After defining the concept of SMI and total ischemia burden, the author approaches the pathophysiology of myocardial ischemia and focuses on the ischemic cascade. Concerning the detection of SMI the importance of exercise testing and Holter ECG is stressed. Following the classification of SMI proposed by P. F. Cohn, the author analyzes SMI type III with particular interest. He refers the prevalence of SMI in patients suffering from chronic stable angina, and focuses on the prognostic importance of SMI. Afterwards, the problem of treatment and prognostic implications is approached. The paper ends with mention of the results of the most important clinical trials in this field: CASIS, CAPE, TIBBS, ASIST, ACIP, TIBET. PMID- 9341024 TI - [Chronic angina pectoris in the patient with arterial hypertension]. AB - Reference is made to the close relationship between high prevalent arterial hypertension and coronary heart disease, including the vascular risk factor as well as the common genetic and psychosocial factors that influence the development and parallel evolution of vascular and heart diseases. The importance of assessing of the total burden of coronary heart disease risk and its general management is emphasized and some aspects of the treatment of angina pectoris in hypertensive patients are mentioned. PMID- 9341025 TI - [Angina pectoris with normal coronary arteries]. AB - The occurrence of angina pectoris in patients with normal coronary angiograms is a subject of investigation since its first description as an entity in 1967. Its etiology remains unknown but, in the absence of disease of the epicardial coronary arteries, dysfunction of the small, most distal vessels and the endothelium appear to be incriminated. In this review, the most recent knowledge and understanding of the pathophysiology, clinical presentation and management of this syndrome is presented. PMID- 9341026 TI - [The medical therapy of angina pectoris]. AB - The angina pectoris crisis should be treated with nitroglycerin S.L.. In our days as in the 19th century, the non pharmacological therapeutic approach for angina pectoris (per se and to improve the free interval between crises) is still the same. This consists of the reduction of mental and physical stress, to stop smoking, improve light exercise, reduce obesity, and control other risk factors for coronary disease. Beta blockers are the choice drugs followed by calcium antagonists and nitrates. It is recommended that no short action calcium antagonists be used. Nitrates must be given with free intervals of action to avoid tachyphylaxis. The anti-platelet therapy can not be forgotten with aspirin or ticlopidine. Patients with refractory angina pectoris should be coronariography for performed eventual revascularization process. PMID- 9341027 TI - [Angina pectoris: the lipid profile and clinical practice]. AB - After a brief historical synopsis of lipid lowering therapy and coronary artery disease, the actual issues were discussed. The problem of secondary prevention of coronary artery disease was addressed on the basis of the recently published studies. A special emphasis was given to the 4S and CARE studies and their results discussed. The problem of primary prevention in the light of the West of Scotland Study is discussed. PMID- 9341028 TI - [Angina pectoris: the possibilities for surgical treatment. Recent progress]. AB - After reviewing the history of coronary surgery the author describes the different grafts used in coronary artery bypass grafting and its results. They vary with coronary anatomy, left ventricle function, age, arterial hypertension, diabetes, etc. The rationale of the different surgical approaches are discussed, emphasizing the advantages of a close medical-surgical cooperation. PMID- 9341029 TI - [Stress]. AB - The general adaptation syndrome is discussed on the light of recent discoveries on hypothalamic peptides and of their possible influence in survival and in induction of diseases. The problem of stress in alcoholism is reviewed. The author ends with a short souvenir of Hans Selye. PMID- 9341030 TI - [Stress tests in old age. The choice of the stress protocol]. AB - OBJECTIVE: To evaluate, in an elderly Portuguese population, the diagnostic capacities of the most popular treadmill stress test protocols. DESIGN: Retrospective study of an elderly Portuguese population submitted to a stress test. PATIENTS AND METHODS: A population of 45 patients (35 male), aged 65 or more years, consecutively submitted to a stress test. The average age of the group was 67.8 +/- 2.9 years. The three protocols (Bruce, Bruce Modified and Naughton) were comparatively studied in terms of the patients capacity to execute the protocol, capacity to obtain a maximum and a diagnostic stress test, and the complications of the stress test protocol. MAIN RESULTS: The Bruce protocol was used in 19 patients, the Bruce Modified in 13 patients and the Naughton protocol in 13 patients. The three protocols did not lead to any complication. The Bruce protocol led to a larger increment in heart rate (p < 0.001) and to larger maximum rate pressure product (p < 0.05) than the Naughton one. The Bruce protocol obtained a larger number of diagnostic tests (p < 0.01) and a significantly lower number of inconclusive stress tests. The Naughton protocol led to a larger duration of the exercise tests and was not suitable for some of the elderly patients owing to the exhaustion of the protocol. The results obtained with the Bruce Modified protocol were among those of other two protocols. CONCLUSIONS: In the elderly, the stress tests are safe and useful in the diagnosis of exercise induced ischemia and in the stratification of cardiovascular risk. From the protocols studied, the Bruce protocol was the most adequate, globally speaking, for this group of patients. The Bruce protocol presented a better diagnostic capacity with no complications related to the protocol. PMID- 9341031 TI - [The effect of left ventricular dysfunction on nocturnal desaturation in patients with chronic emphysematous bronchitis and PaO2 55-70 mmHg]. AB - The possibility of nocturnal oxygen desaturation (NOD) in patients with chronic bronchitis and emphysema (CBE) even with basal hypoxemia greater that 55 mmHg is well recognised. Nocturnal hypoventilation is admitted as the main cause for this NOD. In this study we evaluate how the presence of left ventricular dysfunction (LVD) could aggravate NOD. Thirty-six patients with CBE and basal stabilised PaO2 55-70 mmHg underwent right heart catheterisation and polysomnographic study. NOD was defined as more than 30% of total sleep time with SaO2 less than 90%; LVD was defined as capillary pressure greater than 15 mmHg. Six patients were excluded from analysis because of sleep apnoea syndrome. In the remaining 30 patients (20 men, 10 women; mean age = 65.88.6 years; mean FEV1 = 0.970.31 litres; 43.316.6% predicted; mean basal PaO2 = 61.83.6 mmHg) 8 had LVD and 18 and NOD. Patients with NOD had a greater diurnal level of hypoventilation (basal PaCO2 = 44.63.8 vs. 414.1 mmHg; p = 0.025). Patients with LVD, despite identical diurnal pulmonary function, showed a significantly p < 0.05) greater degree of NOD (mean nocturnal SaO2 = 84.56.4 vs 89.52.5; minimal nocturnal SaO2= 68.517.3 vs. 79.47.8; Time spent with SaO2 < 90% = 78.833.7 vs. 43.138.7). We conclude that the presence of LVD in patients with CBE and PaO2 55-70 mmHg aggravates the intensity and the time spent with NOD, probably because of aggravation of hypoventilation or ventilation/perfusion mismatching. PMID- 9341032 TI - [Coronary artery disease, myocardial perfusion and ventricular function in Q-wave and non-Q-wave myocardial infarcts]. AB - Controversy remains in considering non-Q wave myocardial infarction (NQMI) a distinct pathophysiological entity of Q wave myocardial infarction (QMI). In order to analyze the severity of coronary artery disease, extension of myocardial scar or myocardial ischemia and ventricular function, 78 consecutive patients with QMI and 32 with NQMI, mean age 55.4 +/- 8.5, not submitted to thrombolytic therapy, were studied. Coronary angiography, exercise thallium scintigraphy and radionuclide ventriculography were performed in all at least within 3 months of a prior myocardial infarction. In the present study the occurrence of QMI was significantly more frequent in older patients than NQMI. There was no prevalence of occlusion either in the right, left circumflex or left anterior descending coronary arteries in both groups. Ejection fraction, degree of occlusion and presence of collateral circulation showed an equal prevalence in QMI and NQMI patients. A higher incidence of multivessel disease was found in NQMI that had less necrosis than QMI patients. The prevalence of exercise induced thallium-201 redistribution defects within the infarct zone was substantially higher and involved more scar segments in NQMI patients. Physiological and clinical consequences of coronary thrombosis depends on the size and the number of diseased arteries, the approach the pathophysiologic consequences of coronary disease in terms of fractal structure has been suggested. A pronounced heterogeneity in regional myocardial blood flow in a fractal branching arterial network may be responsible for the pathophysiologic differences of coronary thrombosis between Q-wave and non Q-wave infarction. PMID- 9341033 TI - [Cardiac metastases from a colonic tumor]. AB - Antemortem diagnosis of cardiac metastasis is rarely observed. A case of cardiac metastasis from colon adenocarcinoma is reported. The diagnosis of an intracavitary mass of the right atrium and vegetant mass of the tricuspid valve was possible by two dimensional transthoracic echocardiography. The authors make a brief reference to the clinical and imaging features of secondary cardiac tumors, with special emphasis on echocardiography. PMID- 9341034 TI - [Emerging and spreading infectious diseases]. PMID- 9341035 TI - [Health behaviors in Portugal. Epidemiological studies]. AB - It is necessary to evaluate habitudes and behaviours in health in Portugal. The purpose of this review is to analyse some portuguese epidemiological studies. CINDI-Portugal, ERICA-Portugal and the National Health Inquiry were national studies regarding multiple health data. Nutritional attitudes and practices have been evaluated since 1940. The National Food Study and the Study of Prevalence of Obesity in Portugal studied Portuguese nutritional attitudes. There are studies of the prevalence of eating disorders i school populations. Some research about the association between food and disease is reported. The Portuguese Food Balance is reviewed. Some methodological problems are pointed out and the results discussed. The results of Portuguese studies suggest that our country is in danger of errors in the course of consumption attitudes. PMID- 9341036 TI - [The seroprevalence for hepatitis E viral antibodies in the northern region of Portugal (among the donor population)]. AB - The authors have undertaken a study in the north of Portugal in what concerns the prevalence of antibodies for HEV donors. This study was transverse through observation and survey. A sample of 1.473 donors was randomly selected from among the population of donors of the Oporto Regional Blood Centre. This sample was representative of the northern region and stratified by district. The usual EIA method has been used to search for HEV antibodies. The results, which were repeatedly reactive, have been confirmed by Western Blot. The prevalence of antibodies for Hepatitis E was 2.5%. The test identified a possible cohort effect and the sample showed that the average age of the donors with or without antibodies was identical. No difference was observed regarding sex, distribution by districts or the presence of antibodies for HB virus. Hepatitis E is benign, without chronic state carrier and almost every patient presents acute symptomatology. The value of prevalence found as well as the absence of clinical symptomatology in the observed donors and the normal state of the hepatic markers, indicate that the use of blood and components obtained from this population should not be a problem. The recommendation proposed by the European Council does not mention the systematic research of HEV. However the epidemiological HEV survey is considered important from the transfusional point of view and as an indicator of the sanitary development of the population. Bearing in mind the developing status of Portugal in this area, it is of utmost importance to create the means to reach European standards. PMID- 9341037 TI - [The risk of seroconversion in surgeons of the hepatitis B, hepatitis C and human immunodeficiency viruses (in a specific surgical population)]. AB - The prevalence of hepatitis B surface antigen (HBs Ag) and antibody to hepatitis C virus (HCV) and human immunodeficiency virus (HIV) was determined in the serum specimens of 288 patients treated surgically in the orthopaedic department of an urban public teaching hospital. The cumulative risk of HBV, HCV and HIV seroconversion for an orthopaedic surgeon during the surgical career span was calculated. We found that 1.4%, 3.1% and 1.7% of patients were seropositive for HBsAg, HCV antibody and HIV antibody, respectively. Seropositivity was neither associated with age nor with trauma, whereas male patients had a greater likelihood of seropositivity. Risk factor assessment did not prove to be discriminating in identifying which patients may pose a potential exposure risk. This study supports the concept of universal infection control precautions for orthopaedic surgeons regardless of the patients' risk factor or serologic status. PMID- 9341038 TI - [Nosocomial infection in a pediatric intensive care unit]. AB - Patients in intensive care units (ICU) are 3 to 4 times more prone to nosocomial infection (NI) than patients in general wards owing to the severity of their pathology and the frequent use of invasive procedures. The aim of this study was to establish the incidence of NI in an ICU and the associated risk factors. PATIENTS AND METHODS: During 18 months, all patients with severity scores III and IV (Clinical Classification System) were studied, (n = 575). The admissions were mainly due to accidents (24.7%), neurological (19.1%), surgical (17.2%), respiratory (11.1%) and infectious (7.0%) disease. The mean duration of stay was 2.4 days. The mortality was 5.2%. The evaluation protocol of these patients included determination of the PRISM score, registration of every invasive procedure and daily search for clinical and laboratory signs of infection. NI was defined according to the criteria of the Centers for Disease Control. Both the intrinsic and extrinsic risk factors were analysed and in the statistical analysis the null hypothesis was rejected at the significance level of p < 0.05. RESULTS: The incidence of NI was 7.6%. The infections occurred in the respiratory tract in 20 patients, bloodstream in 8, genito-urinary tract in 2, central nervous system in 2, skin in 2, gastrointestinal tract in 2, eyes in 1 and surgical wound in 1. There were isolates in 60.6%. The mean duration of stay was longer in patients with NI (9.8 versus 1.9). The factors most closely associated with NI were higher PRISM scores, malnutrition, immunodeficiency failure of 2 or more organs, administration of antibiotic since admission or corticosteroids and simultaneous use of 3 or more invasive procedures. The risk of pneumonia was significantly increased in patients with mechanical ventilation and all the patients with bacteremia had central venous catheters. The mortality was higher in the group with NI (18.2%) than in the group without NI (4.4%). CONCLUSIONS: The incidence of NI is acceptable in our ICU. The most frequent location was the respiratory tract (52.6%). The pathogens most frequently isolated in this ICU were Gram negative rods. The risk of NI increased in more debilitated patients with more severe disease who were administered antibiotic or corticosteroids and submitted to more invasive procedures. PMID- 9341039 TI - [Post-occupational exposure HIV infection in health workers: an update and preventive measures]. AB - The author reviews the worldwide references concerning the problem of HIV infection in health care workers (HCW) after occupational exposure in its different aspects: infection risk rate (0.36-0.4%); common types of exposure (percutaneous, contact, and mucomembranous); main risk factors related to the lesion, the patient (donor) and HCW (receptor); and preventive attitude to be adopted. In what concerns prevention, the author emphasises the universal precautions (nosocomial infection prevention) and drug prophylaxis with anti retroviral drugs-reasons for administration, inclusion and exclusion criteria, and the doubts and certainties regarding the risk-benefit, dosage, duration of treatment, and follow-up. Based on current facts, a management guideline, that is in practice in Santa Maria Hospital, is proposed to challenge this controversial condition, including post-exposure drug use and its effect on hospital policy. The future perspective is described, bearing in mind all the facilities available. PMID- 9341040 TI - [Cutaneous infection with the cytomegalovirus virus in AIDS patients]. AB - Cutaneous cytomegalovirus (CMV) infection has been observed in a variety of nonspecific skin lesions. Because of this fact, its diagnosis is rare and frequently accidental. The presence of the virus has also been observed in apparently normal skin, from both a clinical and histological point of view. In this context, skin biopsy and immunohistochemistry are often the first means of diagnosis of systemic CMV infection. In 180 skin biopsies carried out on HIV patients in the Infectious and Parasitic Diseases Unit, typical histopathological findings of CMV infection in a nonspecific skin lesion were observed in only one patient. Although the patient showed no extracutaneous manifestations at this time, she died soon after this diagnosis. Because of this fact, we review the literature and discuss the difficulties and implications of the diagnosis of cutaneous CMV infection in AIDS patients. PMID- 9341041 TI - [The role of general practitioners in the control of HIV infection and in the treatment of seropositive and AIDS patients]. PMID- 9341042 TI - [Acute generalized exanthematous pustulosis due to Coxsackie B4 virus]. PMID- 9341043 TI - [Will acanthosis nigricans be a new cutaneous manifestation of human immunodeficiency virus infection?]. AB - The authors describe a clinical case of AIDS presented by three opportunistic infections (esophageal candidiasis, tuberculosis and atypical mycobacteriosis) and a dermatological manifestation--acanthosis nigricans--not described in medical literature as accompanying those entities. The exclusion of most common etiologies of acanthosis nigricans and its regression following treatment for those infections suggests that with AIDS it behaves like a paraneoplastic syndrome. Screening for HIV antibodies should be the rule whenever this dermatological manifestation is present. PMID- 9341045 TI - [Syphilis in an HIV-infected patient]. AB - We report the case of a 28-year-old female prostitute, intravenous drug user, seropositive for human immunodeficiency virus and with unusual manifestations of lues infection. The presence of plantar keratoderma, alopecia of the scalp, total loss of hair eyebrows and eyelashes with bilateral chorioretinitis is emphasised. Non-treponemal and treponemal tests for syphilis showed reactivity, but with abnormal serologic expression. The possible relationship between H.I.V. infection and the natural course of syphilis is discussed. PMID- 9341044 TI - [Verrucous herpes zoster in AIDS patients]. AB - The authors describe a case of disseminated Herpes-Zoster (VZV) in an HIV 1 positive patient with AIDS. Hyperkeratotic characteristics, acyclovir resistance and sensitivity to foscarnet of cutaneous lesions are the most important features of this example. From the casuistics of the department, the authors describe two similar cases and review the medical literature with emphasis on etiopathogenic, diagnostic and therapeutic factors of lesions caused by DNA Virus in immunocompromised hosts. PMID- 9341046 TI - [Lyell's syndrome in an AIDS patient]. AB - Toxic epidermal necrolysis (TEN), or Lyell's syndrome, is a rare cutaneous reaction, most often drug related with drugs such as sulfonamides, non-steroidal anti-inflammatory drugs, anti-convulsants and allopurinol. It is characterised by extensive mucocutaneous detachment and usually associated with severe constitutional symptoms. The mortality rate is about 20-66%. The incidence of TEN is increasing among HIV-infected patients, mostly those in an advanced stage of the disease. Although the pathophysiology of TEN is still unclear, data have been reported that support different aetiologies: an immune mechanism, an abnormal pattern of drug biotransformation and a modulating genetic susceptibility. The diagnosis of Lyell's syndrome is mainly clinical because the confirmation of the drug involved is more difficult since there are no safe and reliable skin or in vitro tests that are generally available. THe authors report a case of TEN in a patient with AIDS whose clinical characteristics and evolution are a particular example of this pathology and of the related problems of diagnosis and therapeutics. The fact that this patient presented a prior adverse drug reaction (ADR) besides Lyell's syndrome and a later cutaneous hyperreactivity, raised a complex problem: the risk of reintroducing drugs previously withdrawn due to their possible association with ADRs and the absolute need to treat active disorders. In view of the increased risk of severe cutaneous ADRs, namely TEN, in patients with AIDS, the authors point out the need to set up new guidelines concerning both an early and accurate diagnosis and treatment of these situations and the management of subsequent therapeutics. PMID- 9341047 TI - [Tuberculosis: a forgotten challenge]. AB - Tuberculosis continues to be a serious problem in public health even thought eh causative bacillus was discovered over 100 years ago. The authors present a clinical case that illustrates the current concern regarding the disease. The case concerns a child of emigrant parents, who are drug addicts and whose illness has not been fully ascertained. The child was hospitalised at three and an half months with fever, malnutrition, opisthotonos and hepatosplenomegaly. After the diagnosis of disseminated tuberculosis affecting the central nervous system, treatment was started with five antituberculosis drugs and with corticosteroids. Respiration improved favourably, but after 19 days the patient suffered a partial tonic-clonic seizure. Subsequently, hydrocephalus was observed and a shunt was applied. Bacteriological examination of the gastric aspirate showed a strain of Mycobacterium tuberculosis resistant to isoniazid and streptomycin. In the eight month of therapeutics, three antituberculosis drugs were still being administered, the shunt was still present and the patient showed a severe psychomotor retardation. The child belonged to a risk group and presented a serious form of tuberculosis with multidrug-resistance, illustrating that this group of children is particularly vulnerable and reflect transmission of this illness among adults. PMID- 9341048 TI - Proceedings of the XIth International Congress of the World Veterinary Poultry Association. Budapest, Hungary, 18-22 August 1997. PMID- 9341049 TI - Cellular adhesion molecules: regulation and functional significance in the pathogenesis of liver diseases. AB - Adhesion molecules are cell surface glycoproteins that are critical for the localization of leukocytes at sites of inflammation. This review discusses the current knowledges of adhesion molecule expression in normal liver and its upregulation on individual liver cell types during liver inflammation. Cytokines, chemokines, complement factors, and lipid-derived mediators are critical for increased gene transcription and activation of constitutively expressed adhesion molecules. The specific role of selectins, integrins, and members of the immunoglobulin gene superfamily in sinusoidal and venular leukocyte sequestration, transendothelial migration, and adherence to target cells in the liver is described. Increased understanding of these basic mechanisms of communication between resident liver cells and infiltrating leukocytes (neutrophils, lymphocytes, macrophages) not only advances our insight into the pathophysiology of hepatic inflammation but also identifies promising new targets for therapeutic interventions and expands the spectrum of diagnosis and treatment of liver diseases, including alcoholic hepatitis and cirrhosis, viral hepatitis, ischemia-reperfusion injury (transplantation, tumor resection, shock), sepsis- or endotoxin-induced liver injury, acute and chronic rejection, primary biliary cirrhosis, and primary sclerosing cholangitis. PMID- 9341050 TI - Strengthening psychiatry's dedication and commitment to compassionate care, educational excellence, and creative research. PMID- 9341051 TI - Response to the presidential address: new challenges for proven values. PMID- 9341052 TI - Microchannel electrophoretic separations of DNA in injection-molded plastic substrates. AB - Microfabricated electrophoretic separation devices have been produced by an injection-molding process. The strategy for producing the devices involved solution-phase etching of a master template on a silicon wafer, followed by electroforming more durable injection-molding masters in nickel from the silicon master. One of the nickel electroforms was then used to prepare an injection mold insert, from which microchannel chips in an acrylic substrate were mass-produced. The microchannel devices were used to demonstrate high-resolution separations of double-stranded DNA fragments with total run times of less than 3 min. Run-to-run and chip-to-chip reproducibility was good, with relative standard deviation values below 1% for the run-to-run data and in the range of 2-3% for the chip-to chip comparisons. Such devices could lead to the production of low-cost, single use electrophoretic chips suitable for a variety of separation applications, including DNA sizing, DNA sequencing, random primary library screening, and rapid immunoassay testing. PMID- 9341053 TI - Theory of optical chromatography. AB - To evaluate the performance of optical chromatography, a number of equations are theoretically derived using a ray-optics model. These mathematical formalisms are experimentally verified by determining the relationship between the velocity of motion of a polystyrene bead with respect to the intensity of an applied radiation force under the condition where there exists no applied fluid flow. The force is confirmed to be at a maximum at the focal point and to decrease with increasing distance from this position. The radiation force is verified to be proportional to the square of the particle size when the particle diameter is much smaller than the beam diameter. In addition, the radiation force is ascertained to be proportional to the laser power. These results are in excellent agreement with the proposed theoretical model, which is based on ray optics. Furthermore, by analogy with conventional chromatography, fundamental parameters such as retention distance, selectivity, theoretical plate number, and resolution are calculated, and optimum conditions for chromatographic separation are discussed. Based on the results obtained, the dynamic range can be extended by increasing laser power and decreasing flow rate. Peak broadening is primarily caused by variations in laser power and flow rate of the medium for large particles (< 1 microm). It is possible, in theory, to distinguish particles whose diameters differ by less than 1% for particles with a diameter larger than 1 microm. Three sizes of polystyrene beads are well separated at a flow rate of 20 microm s(-1) and a laser power of 700 mW. This technique is also applied to the separation of human erythrocytes. Two fractions, one consisting of cells ranging from 1.5 to 2.4 microm in diameter and another consisting of cells ranging from 3.5 to 5.7 microm in diameter, are observed. Optical chromatography is useful for separation and size measurement of particles and biological cells. PMID- 9341054 TI - Time-lag focusing MALDI time-of-flight mass spectrometry for polymer characterization: oligomer resolution, mass accuracy, and average weight information. AB - We report a polymer characterization study by matrix-assisted laser description/ionization (MALDI) on a linear time-of-flight instrument equipped with pulsed ion extraction for time-lag focusing. It is demonstrated that time lag focusing MALDI provides improved mass resolution and mass accuracy over continuous extraction instruments. Oligomer resolution is extended to a much higher mass range than that observed even by continuous extraction reflectron systems. This allows new opportunities to study the chemical composition and determine the molecular weight of individual components in a mixture of higher molecular weight polymers. It is shown that oligomer resolution can be obtained for poly(ethylene glycol) (repeat unit mass of 44) of mass up to 25,000 u and poly(styrene) (repeat unit mass of 104) up to 55,000 u. Mass measurement accuracy of 80 ppm or better is demonstrated, and the relevance to end-group analysis is shown for two derivative of poly(ethylene glycol) used as slow-release drugs. The analysis of the molecular weight distribution was investigated at several extraction pulse potentials to determine if there was an effect on the relative peak area. We found that the values of the number-average molecular weight (Mn and the weight-average molecular weight (Mw) do not change significantly for a poly(styrene) blend with oligomer masses between 2,000 and 15,000 u and a polydispersity of 1.155. The values are within the 1.6% standard deviation observed for repeat analyses at the same extraction pulse. PMID- 9341055 TI - A predictive morphometric model for the obstructive sleep apnea syndrome. AB - BACKGROUND: Mathematical formulas have been used to clinically predict whether patients will develop the obstructive sleep apnea syndrome (OSAS). However, these models do not take into account the disproportionate craniofacial anatomy that accompanies OSAS independently of obesity. OBJECTIVE: To determine the accuracy of a morphometric model, which combines measurements of the oral cavity with body mass index and neck circumference, in predicting whether a patient has OSAS. DESIGN: 6-month prospective study. SETTING: University-based tertiary referral sleep clinic and research center. PARTICIPANTS: 300 consecutive patients evaluated for sleep disorders for the first time. MEASUREMENTS: Body mass index, neck circumference, and oral cavity measurements were obtained, and a model value was calculated for each patient. Polysomnography was used to determine the number of abnormal respiratory events that occurred during sleep. Sleep apnea was defined as more than five episodes of apnea or hypopnea per hour of sleep. RESULTS: The morphometric model had a sensitivity of 97.6% (95% CI, 95% to 98.9%), a specificity of 100% (CI 92% to 100%), a positive predictive value of 100% (CI, 98.5% to 100%), and a negative predictive value of 88.5% (CI, 77% to 95%). No significant discrepancies were revealed in tests of intermeasurer and test-retest reliability. CONCLUSIONS: The morphometric model provides a rapid, accurate, and reproducible method for predicting whether patients in an ambulatory setting have OSAS. The model may be clinically useful as a screening tool for OSAS rather than as a replacement for polysomnography. PMID- 9341056 TI - Efficacy of automatic continuous positive airway pressure therapy that uses an estimated required pressure in the treatment of the obstructive sleep apnea syndrome. AB - BACKGROUND: Continuous positive airway pressure (CPAP) is effective therapy for the obstructive sleep apnea syndrome (OSAS). Automatic CPAP devices continuously adjust the positive pressure to the required levels. OBJECTIVE: To determine the efficacy of an automatic CPAP machine used with an estimated reference pressure value. DESIGN: A before-and-after, single-blind trial in which patients were randomly allocated to one of three modes of CPAP administration. SETTING: Referral-based sleep center in a public health care institution. PATIENTS: 36 outpatients with OSAS. INTERVENTION: Continuous positive airway pressure was given at a conventional fixed pressure (group 1), automatic CPAP was given at a measured reference pressure (group 2), and automatic CPAP was given at an estimated reference pressure (group 3). In group 1, the effective pressure was determined during a titration sleep study. In groups 2 and 3, the pressure interval was allowed to vary from 4 cm H2O below reference pressure to 3 cm H2O above reference pressure. In group 3, the estimated value of the reference pressure was determined according to individual anthropometric characteristics. MEASUREMENTS: Sleep studies were performed and measurements of diurnal sleepiness were obtained at each visit. RESULTS: Sleep and breathing disorders and hypersomnolence were alleviated similarly in the three groups. The apnea + hypopnea index remained abnormal in one patient in group 3 for whom the reference pressure had been underestimated. A strong negative correlation was found between the percentage of time spent below reference pressure during CPAP and the difference between the effective and estimated pressures. CONCLUSION: Automatic CPAP can be used with an estimated reference pressure without doing a titration sleep study. The positive pressure trend can be used to determine whether treatment failure is caused by an inadequate pressure setting and to determine the amount of pressure to apply. PMID- 9341057 TI - Use of low-dose oral contraceptives and stroke in young women. AB - BACKGROUND: Low-dose oral contraceptives are widely used, but there are limited data on the cerebrovascular risks associated with these medications. OBJECTIVE: To determine whether use of low-dose oral contraceptives influences the risk for stroke. DESIGN: Population-based case-control study. SETTING: Women 18-44 years of age who resided in western Washington State between 1991 and 1995. PARTICIPANTS: Patients with ischemic stroke (n = 60), hemorrhagic stroke (n = 102), and other types of stroke (n = 11) and controls identified through random digit dialing (n = 485). MEASUREMENTS: Details about oral contraceptive use and other risk factors for stroke were obtained through in-person interviews. RESULTS: The estimated incidences of hemorrhagic stroke and ischemic stroke were 6.4 and 4.3 per 100,000 women-years, respectively. Compared with women who had never used oral contraceptives (after adjustment for risk factors for stroke), current users of low-dose oral contraceptives had estimated odds ratios of 0.93 (95% CI, 0.37 to 2.31) for hemorrhagic stroke and 0.89 (CI, 0.27 to 2.94) for ischemic stroke. Compared with past users of oral contraceptives, current users had odds ratios of 1.41 (CI, 0.67 to 2.96) for hemorrhagic stroke and 1.37 (CI, 0.49 to 3.81) for ischemic stroke. For past users compared with never users, the odds ratios were 0.59 (CI, 0.30 to 1.18) for hemorrhagic stroke and 0.57 (CI, 0.25 to 1.32) for ischemic stroke. The odds ratio for hemorrhagic stroke in current users of low-dose oral contraceptives containing norgestrel or levonorgestrel was elevated (3.23 [CI, 1.24 to 8.41]). Among patients with hemorrhagic stroke, the odds ratio for aneurysmal bleeding associated with current use of low-dose oral contraceptives containing norgestrel or levonorgestrel was 4.46 (CI, 1.58 to 12.53). CONCLUSION: The overall risk for stroke and type of stroke was not increased among current users of low-dose oral contraceptives in the study population. Larger studies are needed to clarify both the relation of risk for stroke to past use of oral contraceptives and the possible association between current use of norgestrel-containing oral contraceptives and hemorrhagic stroke. PMID- 9341058 TI - Endoscopic ultrasonography, fine-needle aspiration biopsy guided by endoscopic ultrasonography, and computed tomography in the preoperative staging of non-small cell lung cancer: a comparison study. AB - BACKGROUND: Current methods for detecting mediastinal lymph node involvement with non-small-cell lung cancer can be inaccurate and are often invasive and expensive. OBJECTIVE: To assess the utility of endoscopic ultrasonography, fine needle aspiration biopsy guided by endoscopic ultrasonography, and computed tomography for the detection of metastases to the posterior mediastinal lymph nodes in non-small-cell lung cancer. DESIGN: Prospective preoperative evaluation of the diagnostic operating characteristics of these procedures. SETTING: Referral-based academic medical center. PATIENTS: 130 consecutive patients with non-small-cell lung cancer who were otherwise good surgical candidates. INTERVENTIONS: All patients had initial computed tomography of the chest; those with enlarged nodes were referred for endoscopic ultrasonography. Endoscopic ultrasonography-guided fine-needle aspiration biopsy was done on suspicious contralateral posterior mediastinal or subcarinal lymph nodes identified by ultrasonography. At surgery, lymph nodes were dissected and categorized by location and underwent histopathologic evaluation. RESULTS: 52 patients were ultimately enrolled in the study: Thirty-one had thoracotomy with mediastinal dissection, and 21 had tumors considered unresectable on the basis of preoperative evaluation. Ultrasonography without aspiration biopsy had an overall accuracy of 84% for predicting metastasis to lymph nodes; computed tomography had an accuracy of 49% (P < 0.025). Twenty-four patients had ultrasonography-guided aspiration biopsy; 14 of 24 were ineligible for surgery because cytology showed malignancy. Results of surgical pathology correlated with negative aspiration cytology results in 9 of 10 patients, the one node with false-negative results contained a 2-mm focus of cancer. The accuracy of ultrasonography-guided aspiration biopsy in diagnosing metastasis to lymph nodes was 96%; the results of this test prompted a change in management in 95% of the patients who had the procedure. CONCLUSIONS: Endoscopic ultrasonography alone or with fine-needle aspiration biopsy adds useful diagnostic information in determining metastasis to posterior mediastinal or subcarinal lymph nodes in patients with non-small-cell lung-cancer. These procedures are especially helpful in the preoperative evaluation of patients with suspicious contralateral mediastinal or "bulky" subcarinal nodes. PMID- 9341059 TI - Granulomatous disease in common variable immunodeficiency. AB - BACKGROUND: Granulomatous lesions are occasionally found in the lymphoid or solid organs of patients with common variable immunodeficiency. OBJECTIVE: To examine the clinical and immunologic conditions in patients with common variable immunodeficiency who have granulomas. DESIGN: Case series. SETTING: Large tertiary care medical center. PATIENTS: 17 hypogammaglobulinemic patients with common variable immunodeficiency whose organ or tissue biopsy samples contained noncaseating granulomas. MEASUREMENTS: Results of lymphocyte function tests. RESULTS: Eight of 17 patients had granulomas at some point before hypogammaglobulinemia was diagnosed. Sixteen of the 17 had deficient T-cell proliferation to mitogens. Although 14 patients received standard treatment with intravenous immunoglobulin, they have had substantial illness, including frequent autoimmune disease. CONCLUSIONS: Dysregulated T-cell function or macrophage activation may have been involved in formation of granulomas and increased illness in hypogammaglobulinemic patients with common variable immunodeficiency. Delay in recognition of antibody deficiency may have contributed to the severity of illness in these patients. PMID- 9341060 TI - Peripheral nerves regenerated in familial amyloid polyneuropathy after liver transplantation. AB - BACKGROUND: Liver transplantation holds promise as a treatment for familial amyloid polyneuropathy. OBJECTIVE: To determine whether peripheral nerves regenerate in patients with familial amyloid polyneuropathy after liver transplantation. DESIGN: Case report. SETTING: University hospital in Matsumoto, Japan. PATIENT: A 34 year-old-women with familial amyloid polyneuropathy who had liver transplantation and showed marked clinical improvement 3 years after surgery. MEASUREMENTS: Histopathologic examination and morphometric analysis of biopsy specimens taken from sural nerves. RESULTS: Diffuse fiber loss and amyloid deposits were seen in a biopsy specimen of the left sural nerve obtained before liver transplantation (total number of myelinated fibers, 1326/mm2 of the endoneurial area). In the biopsy specimen of the right sural nerve, which was obtained 3 years after transplantation, amyloid deposits remained but the number of myelinated fibers was markedly increased (total number of myelinated fibers, 4740/mm2). CONCLUSION: Peripheral nerves regenerated in a patient with familial amyloid polyneuropathy after liver transplantation. PMID- 9341061 TI - HLA-B27-associated cardiac disease. AB - PURPOSE: To review the available information about cardiac disease in relation to the immunogenetic marker HLA-B27 and the inflammatory disorders associated with it (seronegative spondyloarthropathies). DATA SOURCES: English-language clinical studies, case reports, and reviews published in 1995 or earlier were identified from a search of the MEDLINE database. These papers, textbooks, and other bibliographic sources were used to identify other relevant publications, enabling collection of references from 1936 through 1995 in a stepwise manner. STUDY SELECTION: Clinical studies, case reports, and topical reviews on the HLA-B27 histocompatibility locus, seronegative spondyloarthropathies, and cardiac disease were selected. DATA EXTRACTION: Information from systemic or hypothesis-driven studies, including those showing clinically and statistically meaningful associations between disorders, and additional relevant information from case reports and reviews is presented. DATA SYNTHESIS: Immunogenetic, histopathologic, clinical, and electrophysiologic evidence was examined to explore the concept of HLA-B27-associated cardiac disease. RESULTS: Relative to the general population (in which the frequency is 6% to 8%), men (but not women) with pacemakers have a significantly increased frequency of HLA-B27. Furthermore, a cardiac syndrome that consists of severe conduction system abnormalities plus aortic regurgitation is associated with HLA-B27, which was present in 67% to 88% of the patients with both of these clinical findings. This proportion of HLA-B27 is similar to that found in patients with ankylosing spondylitis and Reiter disease. Cardiac and aortic tissue both show intimal proliferation of small arteries (obliterative endarteritis) and fibrosis, as seen in tissues adjacent to afflicted joints. Both cardiac conduction abnormalities and aortic regurgitation occur in patients with various HLA-B27-related extracardiac disorders, regardless of the severity of the latter; in about 50% of patients with these cardiac findings, a diagnosis of an HLA-B57-related rheumatic disease has not previously been made. Third-degree (complete) atrioventricular block related to HLA-B57 is located within the atrioventricular node in 95% rather than the expected 20% of cases. CONCLUSIONS: The heart and the joints are both of major importance as targets for the HLA-B57 associated disease process. HLA-B57-related cardiac lesions may be found in the absence of other rheumatologic manifestations. Furthermore, a genetically linked cardiac syndrome has been defined: the combination of conduction system abnormalities and aortic regurgitation. PMID- 9341062 TI - The generalist/cardiovascular specialist: a proposal for a new training track. AB - The economic forces that are reshaping the delivery of health care in the United States have led to intense examination of the appropriate roles for specialists and generalists. Resolving this issue has profound implications for the future of U.S. health care and for the economic health of academic training centers and individual physicians. The issues are particularly intense in cardiovascular care, a field that has had dramatic success in the application of new diagnostic and therapeutic technology and rapid growth in specialist practitioners but is now under pressure to shrink its ranks. A new generalist/cardiovascular specialist training track and a parallel reduction in the number of standard fellowship training positions in cardiovascular disease may be a partial solution. The first 2 years of the proposed 5-year program would consist of training in internal medicine, the final 2 would consist of training in cardiovascular disease, and the middle year would be a flexible combination of the two. Graduates would be Board eligible in internal medicine but would have enhanced competency in cardiovascular disease. This plan may improve the balance between generalists and specialists, improve the quality of primary and specialized cardiovascular care, and strengthen departments of medicine and academic training centers while facing new economic realities. PMID- 9341063 TI - Smallpox: the triumph over the most terrible of the ministers of death. AB - More than 200 years ago, Edward Jenner performed an experiment that laid the foundation for the eradication of smallpox and transformed humankind's fight against disease. Smallpox afflicted humankind as no other disease had don; its persistence and diffusion were without parallel. The disease brought down at least three empires. Generations watched helplessly as their children succumbed to the disease or were disfigured or blinded by it. Attempts were made to contain smallpox by isolating its sufferers and, later, by using variolation with varying degrees of success. However, the definitive solution was not found until Jenner's work was done at the end of the 18th century. Milkmaids who had developed cowpox from contact with cow udders informed Jenner that they were protected from the human form of the disease; he listened to their folk wisdom and raised it to the status of scientific fact. Jenner did not discover vaccination, but he was the first to demonstrate that this technique offered a reliable defense against smallpox. It was also a reliable defense against other illnesses, such as poliomyelitis, measles, and neonatal tetanus, although this was not known in Jenner's lifetime. PMID- 9341064 TI - Tissue is the issue: is endoscopic ultrasonography with or without fine-needle aspiration biopsy in the staging of non-small-cell lung cancer an advance? PMID- 9341065 TI - Coming to grips with large databases. PMID- 9341066 TI - The prison patient. PMID- 9341067 TI - Transesophageal echocardiography-guided cardioversion: going for broke? PMID- 9341068 TI - Amiodarone and thyroid function. PMID- 9341069 TI - Amiodarone and thyroid function. PMID- 9341070 TI - Depression and primary care. PMID- 9341071 TI - Over-the-counter chromium and renal failure. PMID- 9341072 TI - Over-the-counter chromium and renal failure. PMID- 9341073 TI - Over-the-counter chromium and renal failure. PMID- 9341074 TI - Over-the-counter chromium and renal failure. PMID- 9341075 TI - Early detection of prostate cancer. PMID- 9341076 TI - Early detection of prostate cancer. PMID- 9341077 TI - The role of carotid bruit in screening for carotid stenosis. PMID- 9341078 TI - A physician's hamlet. PMID- 9341079 TI - [Biogenic decomposition of bone collagens]. AB - Soil bacteria, which are an always present component of all burial conditions, substantially contribute to the decomposition of bone collagen, i.e. even modifications of preserved bone collagen. Archaeometric data, which were derived from collagen remnants, therefore may always contain diagenetic components which disturb the biological signals and must subsequently be eliminated. To adequately assess probable diagenetic influences one should refer to our model of the biogenic steps of decomposition. The taphonomic experiments and their verifications of archaeological human bones, which were gathered from different origins and of different historical times, were done with histological, biochemical and biophysical methods. Experimentally, as well as through archaeological bone analysis, we were able to identify a "high" molecular product of decomposition, which contains (1) changes in the amino acid composition caused by specific soil conditions and (2) significant deviations of stable nitrogen and carbon isotopes. Due to decomposition procedures, isotope analysis of isolated amino acids indicate rearrangements of collagen fragments. PMID- 9341080 TI - [Serum proteins in human skeletal remains]. AB - The analysis of non-mineralbound, non-collagenous proteins from ancient Peruvian human bones with electrophoretical methods, SDS-polyacrylamide-electrophoresis and isoelectric focusing, results in molecular weight bands on the one hand and a pH-gradient distribution on the other hand, which are perhaps connected with polymorphic serum proteins. A specific protein identification is possible with immunological methods. PMID- 9341081 TI - [A comparison of trace element and isotope analysis in archeological bones (exemplified by a medieval bone series from Weingarten, Germany)]. AB - The well established methods of isotope- and trace element analysis have been used as a reliable approach in archaeometry in order to reconstruct ecological and social parameters. Both methods offer access to palaeoecological information, however, frequently only one of the methods has been applied on the onegiven material. Up to now, isotope data are especially associated to the determination of food webs and palaeocliamtes, i.e. the position of the examined subjects within an ecological context is englightened. On the other hand, trace element analyses reveal knowledge about group specific nutrition and pollution with toxic substances. For the medieval human bone series of Weingarten (Germany), both methods have been applied independently. The results suggest special emphasis of diagenesis for the interpretation of trace element data: diagenesis and its impact on the stability of biological signals as well as consequences for data interpretation, which is dependent on the analysed phase of the material (mineral or gelatine) and its preservation. PMID- 9341082 TI - [Correlations between cribra orbitalia, archeometric findings on the skeleton and habitat conditions]. AB - The interpretation of cribra orbitalia as an isolated skeletal lesion is mainly based on malnutrition, in particular in terms of iron deficiency or vitamin C deficiency. A chronic iron deficiency anemia should result in both a reduced iron content of the skeleton and an underhydroxylation of the amino acid proline with the consequence of a less stable collagen matrix of the bone. The latter holds also for a vitamin C deficiency. Trace element and amino acid analyses were independently applied to subsamples of two early medieval human skeletal series to test whether a relation between cribra orbitalia and skeletal iron content and amino acid hydroxylation was detectable. For one of these series (Nusplingen, district Balingen, Schwabische A1b), habitat specific properties have already been assumed to be responsible for an insufficient iron supply of the inhabitants. Both archaeometric approaches led to a verification of the malnutrition hypothesis, but for advanced states of the skeletal lesion only. Less affected individuals did not reveal any of the expected relationships. Possible adaptive subsistence strategies are discussed which might compensate for such unfavourable living conditions. PMID- 9341083 TI - [Intraindividual element distribution in the distal femur]. AB - Elemental analysis of archaeological bone is a useful tool for intra- and inter population research. Possibly an intraindividual elemental analysis could also reveal individual influences from the personal life history. The distal part of an archaeological femur was prepared in 48 samples (6 heights, 4 locations, compact/spongious bone). The elements Ca, P, Pb, Sr and Zn were measured by atomic-absorption-spectrometry and photometry. Statistical tests were used to study similarities and differences. In addition, a sample was taken from the middle of the diaphysis to compare these results with the concentrations of the distal part of the femur. The results show a specific distribution for each element. Similarities were only found between Ca and P. A likely interpretation of intraindividual element data is not yet possible due to the lack of the knowledge of the intraindividual element distribution. More intraindividual studies like this one are recommended. PMID- 9341084 TI - [Quantitative nutritional reconstruction by trace element data]. AB - Cemeteries without grave goods permit conclusions on the social structure of the underlying population by morphologic skeleton characteristics alone only very restrictedly; though, this social structure may manifest itself in varied nutritional habits, which mirror for their part in the trace element spectra of the skeletal material. A proportional interrelation between Sr-Ca-ratio in food and Sr-Ca-ratio in bone exists, the proportionality factor (observed ratio = OR) being known; the same goes for Ba. This linearity is not given any longer concerning the interrelation between nutritional habits, i.e. the shares of the single food stuffs, and the trace element concentrations of the food and by that of the bone. This makes the use of statistic procedures (cluster analysis) upon the initial trace element data unreliable, so reconstruction of the actual nutrition is necessary. Suitable equation systems do not only supply the wanted individual nutritional habits, but also indicate the metabolism of other trace elements. PMID- 9341085 TI - [Molecular sex determination in skeletal remains of premature and newborn infants of the Aegerten, Switzerland, burial field]. AB - The morphological and morphometrical analyses of skeletal remains usually give reliable access to the gender of mature individuals while the analyses of skeletal remains of immature individuals allocate only 70-90% of the individuals. However, the use of modern techniques like the Polymerase Chain Reaction (PCR) enables a sex identification also if fragmentary material or skeletal remains of children are investigated. The present study reconstructs the sex ratio of about 120 stillborn and neonate individuals of the burial place Aegerten, Switzerland. The morphometrical sex determination of the children suggests a large excess (about 60%) of female individuals. This finding was compared to the molecular sex identification. In order to perform a molecular sex identification aDNA was extracted from bone samples of the stillborn and neonate individuals. A standard phenol/chloroform extraction and a purification with a silica powder were carried out. Finally, the aDNA samples were amplified with a primer system for the amelogenin gene, that is located on the human sex chromosomes. PMID- 9341086 TI - [Detection of DNA single-copy sequences of prehistoric teeth. Site milieu as a factor for preservation of DNA]. AB - DNA-extracts from prehistoric roots of teeth were analyzed by PCR to investigate microsatellite-systems. At the same time the x/y-chromosome specific system Amel A/B was investigated. The samples were collected from different burial conditions of similar age. Factors leading to limited DNA-degradation are given. For the first time reproducible Quadruplex-amplification on single-copy loci of prehistoric tissue was obtained in samples from the Lichtenstein-Cave, Kr. Osterode/Harz. Positive results are explained by low temperature in the burial site. In addition the results show that desiccation allows DNA-preservation but cannot protect the DNA from damage due to microbial origin. Microorganisms can destroy DNA-structures completely. PMID- 9341088 TI - [Classification of isolated skeletal elements using aDNA typing]. AB - Analysis of ancient DNA of material found in the Lichtensteinhohle, a burial site of the Younger Bronze Age has been used for the first time to assign isolated skeletal elements to corresponding individuals. The method involved DNA typing through amplification of five Short Tandem Repeat loci which are also used in forensic genetics for the determination of kinship and identification. From all of the examined bone samples DNA was successfully extracted and amplification by means of Polymerase Chain Reaction could be carried out. For the skeletal elements allelic profiles which are specific for an individual were set up. These profiles made it possible to recognize bones belonging to one individual. Elements which were not from this individual could be excluded with certainty by aDNA analysis. PMID- 9341087 TI - [Reproducibility of aDNA typing]. AB - The reproducibility of short tandem repeat (STR) amplification of aDNA extracts is limited by means of formation of artifacts during PCR. One of these artifacts, the so called allelic-dropout (failure of the amplification of alleles), may result in false-homozygote typing of a sample, if genotyping is based on the analysis of a single amplification product. 30 tooth- and bone samples collected from 17 individuals of three burial sites were investigated in a blind test. Using the polymerase chain reaction (PCR) the two independent segregating STR loci HUMVWA and HUMTH01 were amplified. Corresponding to a mathematical estimation, multiple amplifications of each sample and genelocus were carried out. The results of this genotyping were compared intraindividually. PMID- 9341089 TI - [Determination of kinship by aDNA analysis]. AB - Molecular sex determination and aDNA typings were carried out on a group of five prehistoric individuals. Due to the burial situation and the individual ages the group was assumed to be a family consisting of the parents and three children. DNA was extracted from teeth and bone samples of all skeletons. The aDNA typings based on the PCR amplification of four different microsatellite DNA loci, so called human short tandem repeats (STR). Results of the molecular sex determinations and allele determinations are presented and compared with morphological determinations. Up to now, the individual biological kinship could be verified for three of the individuals. This is the first proof of individual biological kinship on the molecular level for prehistoric individuals. PMID- 9341090 TI - [Prospects in prehistoric anthropology?]. AB - Although Prehistoric Anthropology seems to flourish there are reasons for concern. Innovative developments as e.g. molecular techniques require rethinking of concepts and more staff and money, requirements for which Prehistoric Anthropology seems not to be well prepared to cope with. One way to drive back competition by foreign subjects could be to set up research co-operation. PMID- 9341091 TI - Anaerobic degradation of polycyclic aromatic hydrocarbons and alkanes in petroleum-contaminated marine harbor sediments. AB - Although polycyclic aromatic hydrocarbons (PAHs) have usually been found to persist under strict anaerobic conditions, in a previous study an unusual site was found in San Diego Bay in which two PAHs, naphthalene and phenanthrene, were oxidized to carbon dioxide under sulfate-reducing conditions. Further investigations with these sediments revealed that methylnaphthalene, fluorene, and fluoranthene were also anaerobically oxidized to carbon dioxide in these sediments, while pyrene and benzo[a]pyrene were not. Studies with naphthalene indicated that PAH oxidation was sulfate dependent. Incubating the sediments with additional naphthalene for 1 month resulted in a significant increase in the oxidation of [14C]naphthalene. In sediments from a less heavily contaminated site in San diego Bay where PAHs were not readily degraded, naphthalene degradation could be stimulated through inoculation with active PAH-degrading sediments from the most heavily contaminated site. Sediments from the less heavily contaminated site that had been adapted for rapid anaerobic degradation of high concentrations of benzene did not oxidize naphthalene, suggesting that the benzene- and naphthalene-degrading populations were different. When fuels containing complex mixtures of alkanes were added to sediments from the two sites, there was significant degradation in the alkanes. [14C]hexadecane was also anaerobically oxidized to 14CO2 in these sediments. Molybdate, a specific inhibitor of sulfate reduction, inhibited hexadecane oxidation. These results demonstrate that a wide variety of hydrocarbon contaminants can be degraded under sulfate-reducing conditions in hydrocarbon-contaminated sediments, and they suggest that it may be possible to use sulfate reduction rather than aerobic respiration as a treatment strategy for hydrocarbon-contaminated dredged sediments. PMID- 9341092 TI - [The effect of different high riboflavin supplements during pregnancy and lactation on performance and erythrocyte glutathione reductase activity of rats]. AB - In two experiments with 160 female Sprague-Dawley rats the influence of various dietary riboflavin supplementations during lactation and during pregnancy and lactation were examined on food intake, body mass, reproduction, hematologic profile and the erythrocyte glutathione reductase activity coefficient (EGR-AC). In the first trial rats were fed a semisynthetic, riboflavin-deficient diet, based on casein and corn starch with various riboflavin supplementations during lactation (0, 2, 4, 6, 8, 10, 12, 40, 400, 4000 mg riboflavin/kg diet). In the second experiment the rats received supplements of 1 and 20 mg riboflavin/kg diet, respectively, during pregnancy. After parturition each group was divided into three sub-groups with riboflavin supplementations during lactation of 1, 5 and 20 mg/kg diet, respectively. Both investigations ended at the 14th day of lactation. Food intake was decreased significantly by 25% and 11% in the groups without riboflavin supplementation or 1 mg riboflavin/kg diet. In the same groups body mass was reduced by 11% and 4%, respectively. With regard to the reproduction parameters the riboflavin supply influenced only the litter mass at the 14th day of lactation and only lactational supply was relevant. In both trials the results of the hematologic profile showed no differences. In riboflavin deficiency (0 or 1 mg riboflavin/kg diet, respectively) the EGR-AC was increased significantly to 1.9 and 1.8, respectively. At the supplementation of 4 5 mg riboflavin/kg diet EGR-AC reached a plateau of 1.45, which was not improved by higher supplements. Concerning the whole reproduction cycle (trial II) there was a stronger influence of the actual lactation-supply on EGR-AC, on the other hand a riboflavin deficiency in pregnancy could be compensated only partially by an optimal supply in lactation. Therefore, based on the parameter EGR-AC an optimal riboflavin supply is recommended for each part of the reproduction cycle. By means of EGR-AC also the riboflavin requirement for lactating rats was derived. Feeding a semisynthetic diet (17.4 MJ ME/kg DM, 20.8% crude protein in DM) a supplementation of 5-6 mg riboflavin/kg or a total content of 6-7 mg/kg diet is recommended. PMID- 9341093 TI - [Glycerol supplementation in broiler rations with low crude protein content]. AB - In connection with the utilization of glycerol, which could become available as a by-product of the fuel production from rapeseed the influence of glycerol feeding with rations of low crude protein content was proved. 61 male day old broiler chickens received ad libitum 8 experimental rations based on maize and soybean meal. The experimental design included 3 factors: 15 or 18% CP; supplementation of essential amino acids or not and a content of pure glycerol of 0 or 10%. During the experimental feeding of 23 days body weight gain, feed conversion ratio, N-balance and the intake and excretion of glycerol was obtained. At the end of the trial the utilization of 15N-methionine and the glycerol content of blood plasma, liver and breast muscle was estimated. Due to the low crude protein content of the rations the body weight gain and N-balance was very low (BWG day 1 to 23: 8.6 to 17 g/animal.d; N-balance day 19 to 23: 0.4 to 1.0 g N/animal.d). The supplementation of essential amino acids was the factor with the highest improving effect on the body weight gain, feed conversion ratio and N-balance. Especially at the begin of the trial animals which received 10% glycerol had have an increased feed intake. The excretion of the supplemented glycerol by excreta amounted to 26% of the intake. However, the glycerol content of the rations did not effect body weight gain, feed conversion ratio, N-balance or utilization of 15N-methionine significantly. The assumption of a saving effect on glucoplastic amino acids due to glycerol feeding could not be manifested. In the mean feeding of glycerol elevated the glycerol level in blood plasma in comparison to the basal level up to 23 times (from 0.6 to 13.6 mumol/ml) and in the breast muscle up to 19 times (from 0.4 to 7.5 mumol/g). Nevertheless, these values were lower than the basal level of glycerol in the liver (17.1 to 19.0 mumol/g), which was significantly increased by glycerol feeding to 128%. Regarding the utilization of glycerol as a by-product of the production of renewable fuels it can be concluded that on the basis of the estimated parameters 10% pure glycerol in broiler rations instead of corn starch is without adverse effects. PMID- 9341094 TI - [Determination of the passage of two markers in the gastrointestinal tract of steers using different evaluation methods: effect of the level of nutrition]. AB - This study was conducted to investigate the effects of amount of intake of a mixed diet by steers on the passage of digesta through the reticulorumen and the whole digestive tract. Six ruminally cannulated steers received a mixed diet consisting on average of 43% grass silage, 25% maize silage, 30% concentrate and 2% mineral-vitamin mix in DM. The experimental design was a repeated 3 x 3 Latin square with 21 days periods. The diet was offered twice daily (07.00 and 19.00 h) at 1, 1.5 and 2 times of estimated maintenance energy requirements. At the beginning of each period, animals received pulse doses of Titanium(IV)-oxide (TiO2) per os and Cr-EDTA intraruminally. Following marker administration, faecal marker concentrations were determined over 120 h. Passage parameters were estimated by a mono-exponential and a bi-exponential model, by models including gamma age-dependency and by a model calculating total tract mean retention time as mean value of all points on the marker-excretion curve. Passage rate of TiO2 from the reticulorumen increased with higher intakes, whereas mean retention time in the whole tract decreased. In general, results of different models were consistent across intakes. Values for passage of TiO2 were in good agreement with those reported for small particles, when similar diets were fed to cattle. Rate of passage of Cr-EDTA from the reticulorumen increased with higher intakes, and mean retention times of Cr-EDTA in the reticulorumen and in the whole tract decreased. Differences between models with or without age dependency were greater for Cr-EDTA than for TiO2. Fit to Cr-EDTA excretion curves was not satisfactory for models with gamma age dependency. Irrespective of model and marker, passage from the reticulorumen accelerated markedly, whereas retention times in the reticulorumen and in the whole tract decreased, as intake increased from maintenance energy requirements to 1.5 times maintenance. PMID- 9341096 TI - 10th National Congress of the Italian Society for the Study of Atherosclerosis. Rome, Italy, 2-3 December 1996. Abstracts. PMID- 9341095 TI - [Planning, realization and evaluation of post-marketing surveillance studies. Recommendations of the Society for Phytotherapy]. AB - Post-marketing-surveillance studies with herbal drugs usually are prospective prescription-epidemiological studies, which should allow statements on quality, efficacy and safety. Until now neither laws nor concrete normative guidelines for the methodology and the evaluation of post-marketing-surveillance studies are existing which could be used for pharmacovigilance. In the present paper guidelines for planning, realisation and evaluation are presented which should allow studies of high quality. The essential components required for the investigational plan are focussed. Also recommendations on the obligatory, optional and special components of the study protocols are made. Additionally statistical methods which allow the evaluation of the therapeutic efficacy are presented. PMID- 9341097 TI - The relevance of mechanistic data to the interpretation and extrapolation to humans of rodent carcinogenicity data. PMID- 9341098 TI - alpha2u-Globulin nephropathy and ravens: do ravens of a different feather flock together? PMID- 9341101 TI - Trouble in paradise. PMID- 9341099 TI - Doubting nongenotoxic mechanisms of renal cancer: comparing apples and oranges in the alpha2u-globulin hypothesis. PMID- 9341100 TI - Weight of evidence versus weight of speculation to evaluate the alpha2u-globulin hypothesis. PMID- 9341102 TI - Atomic legacy in the Marshall Islands. PMID- 9341104 TI - The road to justice. PMID- 9341103 TI - Solvent solution. PMID- 9341105 TI - Nucleosome transactions on the promoters of the yeast GAL and PHO genes. PMID- 9341106 TI - Ubc9p is the conjugating enzyme for the ubiquitin-like protein Smt3p. AB - At least one essential function of Smt3p, a Saccharomyces cerevisiae ubiquitin like protein similar to the mammalian protein SUMO-1, involves its posttranslational covalent attachment to other proteins. Using Smt3p affinity chromatography, we have isolated the second enzyme of the Smt3p conjugation pathway and have found that it is identical to Ubc9p, a previously identified protein that has extensive sequence similarity to the ubiquitin-conjugating enzymes (E2s) and that is required for yeast to progress through mitosis. A hallmark of E2s is the ability to form a thioester bond-containing covalent intermediate with ubiquitin (Ub). While we were unable to detect formation of a Ub approximately Ubc9p thioester, Ubc9p was found to form a thioester with Smt3p, indicating that Ubc9p is the functional analog of E2s in the Smt3p pathway and that this step is distinct from the ubiquitin pathway. Ubc9p is required for attachment of Smt3p to other proteins in vitro, suggesting that it is the only such enzyme in S. cerevisiae. These results suggest that, like ubiquitination, Smt3p conjugation may be a critical modification in cell cycle regulation. PMID- 9341108 TI - Activation of the HIV-1 long terminal repeat by nerve growth factor. AB - The brain is an important target for the human immunodeficiency virus type 1 (HIV 1). We show here that nerve growth factor (NGF), which induces neuronal differentiation and survival, causes a strong activation of the HIV-1 long terminal repeat by a Ras/Raf-dependent mechanism in PC12 cells. Mutation of the kappaB sequences contained whithin the long terminal repeat reduces NGF-mediated stimulation. NGF does not activate NF-kappaB in PC12 cells, but rather increases binding of other nuclear factors to the kappaB sequences. Furthermore, a nuclear receptor response element contributes to the stimulatory effect of NGF. The retinoids receptors have been identified as components of the nuclear binding to the nuclear receptor response element in NGF-treated PC12 cells. These results reveal the importance of neurotrophins and nuclear receptor signaling pathways as specific activators of HIV-1 gene expression in neural cells. PMID- 9341107 TI - Heat shock factor 1 represses Ras-induced transcriptional activation of the c-fos gene. AB - Heat shock factor 1, the critical molecular regulator of the stress response is conserved throughout eukaryotic organisms and activates the transcription of heat shock genes. We now show that heat shock factor 1 inhibits the expression of c fos, an immediate early gene that controls responses to extracellular stimuli for growth and differentiation. Heat shock factor 1 inhibits the transcription of the c-fos gene and antagonizes the activating effects of the signal transducing protein Ras on the c-fos promoter and on the promoter of another Ras responsive gene uPA. This property was specific for heat shock factor 1; c-fos repression was not seen with the structurally related protein heat shock factor 2. Repression involved different molecular mechanisms compared with those involved in transcriptional activation by heat shock factor 1 and specifically did not require binding to the c-fos promoter. Thus, in addition to its known role as a transcriptional activator of the cellular heat shock response, heat shock factor 1 also antagonizes the expression of Fos, a key component of the ubiquitous AP-1 transcription factor complex and as such could influence multiple aspects of cell regulation. PMID- 9341109 TI - Molybdenum cofactor biosynthesis. The plant protein Cnx1 binds molybdopterin with high affinity. AB - The molybdenum cofactor is an essential part of all eukaryotic molybdoenzymes. It is a molybdopterin (MPT) revealing the same core structure in all organisms. The plant protein Cnx1 from Arabidopsis thaliana is involved in the multi-step biosynthesis of molybdenum cofactor. Previous studies (Stallmeyer, B., Nerlich, A., Schiemann, J., Brinkmann, H., and Mendel, R. R. (1995) Plant J. 8, 751-762) suggested a function of Cnx1 in a late step of cofactor biosynthesis distal to the formation of MPT, i.e. conversion of MPT into molybdenum cofactor. Here we present the first biochemical evidences confirming this assumption. The protein Cnx1 consists of two domains (E and G) homologous to two distinct Escherichia coli proteins involved in cofactor synthesis. Binding studies with recombinantly expressed and purified Cnx1 and with its single domains revealed a high affinity of the G domain to MPT (kD = 0.1 microM) with equimolar binding. In contrast, the E domain of Cnx1 binds MPT with lower affinity (kD = 1.6 microM) and in a cooperative manner (nH = 1. 5). The entire Cnx1 showed a tight and cooperative MPT binding. Based on these data providing a common link between both domains that matches the previous characterization of plant and bacterial Cnx1 homologous mutants, we present a model for the function of Cnx1. PMID- 9341110 TI - Induction and activation of mitogen-activated protein kinases of human lymphocytes as one of the signaling pathways of the immunomodulatory effects of morphine sulfate. AB - Morphine sulfate causes immunomodulatory and immunosuppressive effects in human. In this study, the signaling pathway involved in these morphine effects was studied. Addition of morphine sulfate to human CEMx174 lymphocytic cells resulted in increased expression of mitogen-activated protein kinase cascade proteins. Morphine enhanced the cellular levels of ERK1 (44 kDa), ERK2 (42 kDa), a 54-kDa ERK, MEK1 (45 kDa), and MEKK (78 kDa). A time-dependent increase in the activated (Thr and Tyr dually phosphorylated) state of ERK1 and ERK2 was also observed. Naloxone, a morphine antagonist, reversed the observed morphine effects, implicating a micro opioid receptor-mediated process. These findings suggest that mitogen-activated protein kinases are important intermediates in signal transduction pathways initiated by morphine receptors in immune cells. PMID- 9341111 TI - Designed disulfide between N-terminal domains of lactose repressor disrupts allosteric linkage. AB - Substitution of Cys for Val at position 52 of the lac repressor was designed to permit disulfide bond formation between the two N-terminal DNA binding domains that comprise an operator DNA binding site. This position marks the closest approach of these domains based on the x-ray crystallographic structures of the homologous purine holorepressor-operator complex and lac repressor-operator complex (Schumacher, M. A., Choi, K. Y., Zalkin, H., and Brennan, R. G. (1994) Science 266, 763-770; Lewis, M., Chang, G., Horton, N.C., Kercher, M. A., Pace, H. C., Schumacher, M. A., Brennan, R. G., and Lu, P. (1996) Science 271, 1247 1254). The V52C mutation was generated by site-specific methods, and the mutant protein was purified and characterized. In the reduced form, V52C bound operator DNA with slightly increased affinity. Exposure to oxidizing conditions resulted in disulfide bond formation, and the oxidized protein bound operator DNA with approximately 6-fold higher affinity than wild-type protein. Inducer binding for both oxidized and reduced forms of V52C was comparable to wild-type lac repressor. In the presence of inducer, the reduced protein exhibited wild-type, diminished DNA binding. In contrast, DNA binding for the oxidized form was unaffected by inducer, even at 1 mM. Thus, the formation of the designed disulfide between Cys52 side chains within each dimer renders the protein operator complex unresponsive to sugar binding, presumably by disrupting the allosteric linkage between operator and inducer binding. PMID- 9341112 TI - Involvement of a specific metal ion in the transition of the hammerhead ribozyme to its catalytic conformation. AB - Previous crystallographic and biochemical studies of the hammerhead ribozyme suggest that a metal ion is ligated by the pro-Rp oxygen of phosphate 9 and by N7 of G10.1 and has a functional role in the cleavage reaction. We have tested this model by examining the cleavage properties of a hammerhead containing a unique phosphorothioate at position 9. The Rp-, but not Sp-, phosphorothioate reduces the cleavage rate by 10(3)-fold, and the rate can be fully restored by addition of low concentrations of Cd2+, a thiophilic metal ion. These results strongly suggest that this bound metal ion is critical for catalysis, despite its location approximately 20 A from the cleavage site in the crystal structure. Analysis of the concentration dependence suggests that Cd2+ binds with a Kd of 25 microM in the ground state and a Kd of 2.5 nM in the transition state. The much stronger transition state binding suggests that the P9 metal ion adopts at least one additional ligand in the transition state and that this metal ion may participate in a large scale conformational change that precedes hammerhead cleavage. PMID- 9341113 TI - Polyunsaturated fatty acid suppression of hepatic fatty acid synthase and S14 gene expression does not require peroxisome proliferator-activated receptor alpha. AB - Dietary polyunsaturated fatty acids (PUFA) induce hepatic peroxisomal and microsomal fatty acid oxidation and suppress lipogenic gene expression. The peroxisome proliferator-activated receptor alpha (PPARalpha) has been implicated as a mediator of fatty acid effects on gene transcription. This report uses the PPARalpha-deficient mouse to examine the role of PPARalpha in the PUFA regulation of mRNAs encoding hepatic lipogenic (fatty acid synthase (FAS) and the S14 protein (S14)), microsomal (cytochrome P450 4A2 (CYP4A2)), and peroxisomal (acyl CoA oxidase (AOX)) enzymes. PUFA ingestion induced mRNAAOX (2.3-fold) and mRNACYP4A2 (8-fold) and suppressed mRNAFAS and mRNAS14 by >/=80% in wild type mice. In PPARalpha-deficient mice, PUFA did not induce mRNAAOX or mRNACYP4A2, indicating a requirement for PPARalpha in the PUFA-mediated induction of these enzymes. However, PUFA still suppressed mRNAFAS and mRNAS14 in the PPARalpha deficient mice. Studies in rats provided additional support for the differential regulation of lipogenic and peroxisomal enzymes by PUFA. These studies provide evidence for two distinct pathways for PUFA control of hepatic lipid metabolism. One requires PPARalpha and is involved in regulating peroxisomal and microsomal enzymes. The other pathway does not require PPARalpha and is involved in the PUFA mediated suppression of lipogenic gene expression. PMID- 9341114 TI - Chloroquine extends the lifetime of the activated insulin receptor complex in endosomes. AB - Insulin signal transduction, initiated by binding of insulin to its receptor at the plasma membrane, activates the intrinsic receptor tyrosine kinase and leads to internalization of the activated ligand-receptor complex into endosomes. This study addresses the role played by the activated insulin receptor within hepatic endosomes and provides evidence for its central role in insulin-stimulated events in vivo. Rats were treated with chloroquine, an acidotrophic agent that has been shown previously to inhibit endosomal insulin degradation, and then with insulin. Livers were removed and fractionated by density gradient centrifugation to obtain endosomal and plasma membrane preparations. Chloroquine treatment increased the amount of receptor-bound insulin in endosomes at 2 min after insulin injection by 93% as determined by exclusion from G-50 columns and by 90% as determined by polyethylene glycol precipitation (p < 0.02). Chloroquine treatment also increased the insulin receptor content of endosomes after insulin injection (integrated over 0-45 min) by 31% when compared with controls (p < 0.05). Similarly, chloroquine increased both insulin receptor phosphotyrosine content and its exogenous tyrosine kinase activity after insulin injection (64%; p < 0.01 and 96% and p < 0. 001, respectively). In vivo chloroquine treatment was without any observable effect on insulin binding to plasma membrane insulin receptors, nor did it augment insulin-stimulated receptor autophosphorylation or kinase activity in the plasma membrane. Concomitant with its effects on endosomal insulin receptors, chloroquine treatment augmented insulin-stimulated incorporation of glucose into glycogen in diaphragm (p < 0.001). These observations are consistent with the hypothesis that chloroquine-dependent inhibition of endosomal insulin receptor dissociation and subsequent degradation prolongs the half-life of the active endosomal receptor and potentiates insulin signaling from this compartment. PMID- 9341115 TI - Transcriptional inhibition by Stat5. Differential activities at growth-related versus differentiation-specific promoters. AB - Prolactin (PRL) induces transcriptional activation of not only growth-related genes such as interferon regulatory factor-1 (IRF-1) but also differentiation specific genes such as beta-casein through a signaling cascade consisting of Janus kinases and Stat (signal transducer and activator of transcription) factors. To understand better the role of Stats in PRL signaling, we cloned rat Stat5b from a PRL-responsive T cell line Nb2. A Stat5b-specific peptide antibody was generated. In PRL receptor reconstituted COS cells cotransfected with Stat5b or Stat5a, both Stat5 proteins become tyrosine phosphorylated and bind to the IRF 1 GAS (interferon-gamma activation sequence) element in a PRL-inducible manner. Unexpectedly, both Stat5b and Stat5a inhibit PRL induction of the IRF-1 promoter, but they mediate PRL stimulation of the beta-casein promoter. Stat5-mediated inhibition was observed only at the native IRF-1 promoter and not at the isolated IRF-1 GAS element linked to a heterologous thymidine kinase promoter. Mutational analyses showed that the DNA binding activity of Stat5b is not required, but the carboxyl-terminal transactivation domain is essential for Stat5b to inhibit PRL induction of the IRF-1 promoter. These results suggest that Stat5b mediates inhibition via protein-protein interactions. In contrast, both DNA binding and transactivation domains of Stat5b are required to mediate PRL induction of the beta-casein promoter. Furthermore, a carboxyl-terminal truncated dominant negative Stat5b can reverse Stat5b inhibition at the IRF-1 promoter. These studies suggest that Stat proteins can act as not only positive but also negative regulators of gene transcription. Further, Stat5 can modulate gene expression without binding to DNA but via protein-protein interactions. PMID- 9341116 TI - Inactivation of eIF2B and phosphorylation of PHAS-I in heat-shocked rat hepatoma cells. AB - Various factors are involved in the heat shock-induced inhibition of protein synthesis. Changes upon heat shock in phosphorylation, leading to inactivation, of eukaryotic initiation factors (eIFs) eIF2 and eIF4E have been shown for several cell types. However, in mammalian cells these changes occur at temperatures of 43 degrees C or higher while protein synthesis is already affected at milder heat shock temperatures. In searching for the cause for the inhibition of protein synthesis, the regulation of eIF2 and eIF4E by additional factors was analyzed. In this respect, the activity of eIF2B was measured during and after heat shock. A very clear correlation was found between the activity of this guanine exchange factor and the levels of protein synthesis, also at mild heat shock conditions. Changes in the phosphorylation of eIF4E and of the eIF4E binding protein PHAS-I were also analyzed. Surprisingly, in H35 cells as well as in some other cell lines, PHAS-I phosphorylation was increased by heat shock, whereas in others it was decreased. Therefore, decreasing the eIF4E availability under stressful conditions does not seem to be a general mechanism to inhibit protein synthesis by heat shock. Regulation of eIF2B activity appears to be the main mechanism to control translation initiation after heat shock at mild temperatures. PMID- 9341117 TI - Tyrosine phosphorylation and relocation of SHIP are integrin-mediated in thrombin stimulated human blood platelets. AB - The SH2 domain-containing inositol 5-phosphatase, SHIP, known to dephosphorylate inositol 1,3,4,5-tetrakisphosphate and phosphatidylinositol 3,4,5-trisphosphate has recently been shown to be expressed in a variety of hemopoietic cells. This 145-kDa protein is induced to associate with Shc by multiple cytokines and may play an important role in the negative regulation of immunocompetent cells mediated by FcgammaRIIB receptor. We report here that SHIP is present in human blood platelets and may be involved in platelet activation evoked by thrombin. Platelet SHIP was identified by Western blotting as a single 145-kDa protein. Both phosphatidylinositol 3,4,5-trisphosphate and inositol 1,3,4, 5 tetrakisphosphate 5-phosphatase activities could be demonstrated in anti-SHIP immunoprecipitates of platelet lysate. Thrombin stimulation induced a tyrosine phosphorylation of SHIP, this effect being prevented if platelets were not shaken or if RGD-containing peptides were present, indicating an aggregation-dependent, integrin-mediated event. Moreover, although the intrinsic phosphatase activity of SHIP did not appear to be significantly increased, tyrosine-phosphorylated SHIP was relocated to the actin cytoskeleton upon activation in an aggregation- and integrin engagement-dependent manner. Finally, the striking correlation observed between phosphatidylinositol 3,4-bisphosphate production and the tyrosine phosphorylation of SHIP, as well as its relocation to the cytoskeleton upon thrombin stimulation, suggest a role for SHIP in the aggregation-dependent and GpIIb-IIIa-mediated accumulation of this important phosphoinositide. PMID- 9341118 TI - Site-specific mutagenesis of a recombinant anti-single-stranded DNA Fab. Role of heavy chain complementarity-determining region 3 residues in antigen interaction. AB - The heavy chain complementarity-determining region 3 (HCDR3) of the anti oligo(dT) recombinant antibody fragment, DNA-1, contributes significantly to antigen binding (Komissarov, A. A., Calcutt, M. J., Marchbank, M. T., Peletskaya, E. N., and Deutscher, S. L. (1996) J. Biol. Chem. 271, 12241-12246). In the present study, the role of separate HCDR3 residues of DNA-1 in interaction with oligo(dT) was elucidated. Based on a molecular model of the combining site, residues at the base (Arg98 and Asp108) and in the middle (Tyr101-Arg-Pro-Tyr Tyr105) of HCDR3 were predicted to support the loop conformation and directly contact the ligand, respectively. Twenty-five site-specific mutants were produced as hexahistidine-tagged proteins, purified, and examined for binding to (dT)15 using two independent methods. All mutations in the middle of HCDR3 led to either abolished or diminished affinity. Tyr101 likely participates in hydrogen bonding, while Tyr104 and Tyr105 may be involved in aromatic-aromatic interactions with the ligand. The residues Arg102 and Pro103 were not as critical as the tyrosines. It is speculated that HCDR3 interacts with the thymines, rather than the phosphates, of the ligand. A 3-fold increase in affinity was observed by mutation of Asp108 to alanine. The highly conserved Arg98 and Asp108 do not appear to form a salt bridge. PMID- 9341119 TI - A 13C nuclear magnetic resonance investigation of the metabolism of leucine to isoamyl alcohol in Saccharomyces cerevisiae. AB - The metabolism of leucine to isoamyl alcohol in yeast was examined by 13C nuclear magnetic resonance spectroscopy. The product of leucine transamination, alpha ketoisocaproate had four potential routes to isoamyl alcohol. The first, via branched-chain alpha-keto acid dehydrogenase to isovaleryl-CoA with subsequent conversion to isovalerate by acyl-CoA hydrolase operates in wild-type cells where isovalerate appears to be an end product. This pathway is not required for the synthesis of isoamyl alcohol because abolition of branched-chain alpha-keto acid dehydrogenase activity in an lpd1 disruption mutant did not prevent the formation of isoamyl alcohol. A second possible route was via pyruvate decarboxylase; however, elimination of pyruvate decarboxylase activity in a pdc1 pdc5 pdc6 triple mutant did not decrease the levels of isoamyl alcohol produced. A third route utilizes alpha-ketoisocaproate reductase (a novel activity in Saccharomyces cerevisiae) but with no role in the formation of isoamyl alcohol from alpha hydroxyisocaproate because cell homogenates could not convert alpha hydroxyisocaproate to isoamyl alcohol. The final possibility was that a pyruvate decarboxylase-like enzyme encoded by YDL080c appears to be the major route of decarboxylation of alpha-ketoisocaproate to isoamyl alcohol although disruption of this gene reveals that at least one other unidentified decarboxylase can substitute to a minor extent. PMID- 9341120 TI - Cyclic AMP-mediated inhibition of angiotensin II-induced protein synthesis is associated with suppression of tyrosine phosphorylation signaling in vascular smooth muscle cells. AB - In the present study, we have examined the effect of increased cyclic AMP (cAMP) levels on the stimulatory action of angiotensin II (Ang II) on protein synthesis. Treatment with cAMP-elevating agents potently inhibited Ang II-induced protein synthesis in rat aortic smooth muscle cells and in rat fibroblasts expressing the human AT1 receptor. The inhibition was dose-dependent and was observed at all concentrations of the peptide. To explore the mechanism of cAMP action, we have analyzed the effects of forskolin and 3-isobutyl-1-methylxanthine on various receptor-mediated responses. Elevation of cAMP did not alter the binding properties of the AT1 receptor and did not interfere with the activation of phospholipase C or the induction of early growth response genes by Ang II. Likewise, Ang II-dependent activation of the mitogen-activated protein kinases ERK1/ERK2 and p70 S6 kinase was unaffected by cAMP. In contrast, we found that increased concentration of cAMP strongly inhibited the stimulatory effect of Ang II on protein tyrosine phosphorylation. Specifically, cAMP abolished Ang II induced tyrosine phosphorylation of the focal adhesion-associated protein paxillin and of the tyrosine kinase Tyk2. These results identify a novel mechanism by which the cAMP signaling system may exert growth-inhibitory effects in specific cell types. PMID- 9341121 TI - The C terminus of cardiac troponin I is essential for full inhibitory activity and Ca2+ sensitivity of rat myofibrils. AB - Although the C terminus of troponin I is known to be important in myofilament Ca2+ regulation in skeletal muscle, the regulatory function of this region of cardiac troponin I (cTnI) has not been defined. To address this question, the following recombinant proteins were expressed in Escherichia coli and purified: mouse wild-type cTnI (WT cTnI; 211 residues), cTnI-(1-199) (missing 12 residues), cTnI-(1-188) (missing 23 residues), and cTnI-(1-151) (missing 60 residues). The inhibitory activity of cTnI and the mutants was tested in myofibrils, from which cTnI.cTnC was extracted by exchanging endogenous cardiac troponin with exogenous cTnT causing the Ca2+ sensitivity of the myofibrils to be lost. Addition of increasing amounts of exogenous WT cTnI or cTnI-(1-199) to cTnT-treated myofibrils at pCa 8 caused a concentration-dependent inhibition of the maximum ATPase activity. However, cTnI-(1-188) and cTnI-(1-151) inhibited this activity to about 75% and 50% of that of the WT cTnI, respectively. We also formed a complex of either WT cTnI or each of the mutants with cTnC, reconstituted the complex into the cTnT-treated myofibrils, and measured the Mg2+-ATPase activity as a function of pCa. We found that the cTnI-(1-188).cTnC complex only partially restored Ca2+ sensitivity, whereas the cTnI-(1-151).cTnC complex did not restore any Ca2+ sensitivity. Each cTnI C-terminal deletion mutant was able to bind to cTnC, as shown by urea-polyacrylamide gel-shift analysis and size exclusion chromatography. Each mutant also co-sedimented with actin. Our results indicate that residues 152-199 (C-terminal to the inhibitory region) of cTnI are essential for full inhibitory activity and Ca2+ sensitivity of myofibrillar ATPase activity in the heart. PMID- 9341122 TI - Expression and secondary structure determination by NMR methods of the major house dust mite allergen Der p 2. AB - There exists a strong correlation between asthma and sensitization to indoor allergens. This study reports on the secondary structure of the major house dust mite allergen Der p 2, determined using heteronuclear NMR methods. The DNA was subcloned from the yeast expression vector pSAY1 into the high yield bacterial expression vector pET21a, resulting in yields of 50 mg/liter. The recombinant protein was shown to have immunoreactivity comparable with that of the natural mite protein using competitive inhibition enzyme-linked immunosorbent assay (ELISA) and a modified monoclonal radioallergosorbent test (RAST). The secondary structure was determined by examining chemical shifts, short and long range NOESYs, JHN-HA coupling constants, and amide exchange rates. From these data, it is clear that Der p 2 is composed of beta-sheets and random coil. Based on long range distance constraints, a number of beta-strands were aligned into two three stranded, anti-parallel beta-sheets. PMID- 9341123 TI - Human homologue of the Drosophila discs large tumor suppressor binds to p56lck tyrosine kinase and Shaker type Kv1.3 potassium channel in T lymphocytes. AB - Human homologue of the Drosophila discs large tumor suppressor protein (hDlg) belongs to a newly discovered family of proteins termed MAGUKs that appear to have structural as well as signaling functions. Consistent with the multi-domain organization of MAGUKs, hDlg consists of three copies of the PDZ (PSD-95/Discs large/zO-1) domain, an SH3 motif, and a guanylate kinase-like domain. In addition, the hDlg contains an amino-terminal proline-rich domain that is absent in other MAGUKs. To explore the role of hDlg in cell signaling pathways, we used human T lymphocytes as a model system to investigate interaction of hDlg with known tyrosine kinases. In human T lymphocyte cell lines, binding properties of hDlg were studied by immunoprecipitation, immunoblotting, and immune complex kinase assays. Our results show that protein tyrosine kinase activity is associated with the immunoprecipitates of hDlg. Immunoblotting experiments revealed that the immunoprecipitates of hDlg contain p56lck, a member of the Src family of tyrosine kinases. The specificity of the interaction is demonstrated by the lack of p59fyn tyrosine kinase and phosphotidylinositol 3-kinase in the hDlg immunoprecipitates. Direct interaction between hDlg and p56lck is demonstrated using glutathione S-transferase fusion proteins of hDlg and recombinant p56lck expressed in the baculovirus-infected Sf9 cells. The p56lck binding site was localized within the amino-terminal segment of hDlg containing proline-rich domain. In addition, we show in vivo association of hDlg with Kv1.3 channel, which was expressed in T lymphocytes as an epitope-tagged protein using a vaccinia virus expression system. Taken together, these results provide the first evidence of a direct interaction between hDlg and p56lck tyrosine kinase and suggest a novel function of hDlg in coupling tyrosine kinase and voltage-gated potassium channel in T lymphocytes. PMID- 9341125 TI - UDP-glucuronosyltransferase, the role of the amino terminus in dimerization. AB - UDP-glucuronosyltransferases (UGTs) comprise an important enzyme system in mammals that is involved in detoxification of a variety of small hydrophobic compounds of both endogenous and exogenous origin. Some evidence suggests that these enzymes may function as oligomers; however, little is known about the domain of interaction or the mechanism of oligomerization. In this work, evidence for a functional dimerization between UGTs is provided by studies on mutated forms of UGT2B1. When two inactive forms of UGT2B1 were co-expressed in cell culture, catalytic activity was restored, indicating that UGT2B1 forms functional dimers. To delineate the dimerization domain, inactive fusion proteins containing the amino- or carboxyl-terminal domains of UGT2B1 were generated and expressed with active UGT2B1. Expression of a fusion protein containing only the amino terminal half of UGT2B1 with active UGT2B1 caused a reduction in UGT2B1 catalytic activity. This reduction in activity was not observed when UGT2B1 was co expressed with a fusion protein containing only the carboxyl-terminal half of UGT2B1, strongly suggesting that the amino-terminal domain is involved in dimerization. Truncation of the immediate amino terminus of UGT2B1 abolished UGT2B1 activity and dimer formation. Activity was also abolished by an L4R substitution in this region of the mature protein, which is highly conserved in the UGT family. These results indicate that UGTs can interact through their amino terminal domains to form catalytically active dimers. Possible mechanisms resulting in the formation and stabilization of the UGT2B1 dimer are discussed. PMID- 9341124 TI - Mapping of a defined neurocan binding site to distinct domains of tenascin-C. AB - Neurocan is a member of the aggrecan family of proteoglycans which are characterized by NH2-terminal domains binding hyaluronan, and COOH-terminal domains containing C-type lectin-like modules. To detect and enhance the affinity for complementary ligands of neurocan, the COOH-terminal neurocan domain was fused with the NH2-terminal region of tenascin-C, which contains the hexamerization domain of this extracellular matrix glycoprotein. The fusion protein was designed to contain the last downstream glycosaminoglycan attachment site and was expressed as a proteoglycan. In ligand overlay blots carried out with brain extracts, it recognized tenascin-C. The interaction was abolished by the addition of EDTA, or TNfn4,5, a bacterially expressed tenascin-C fragment comprising the fourth and fifth fibronectin type III module. The fusion protein directly reacted with this fragment in ligand blot and enzyme-linked immunosorbent assay procedures. Both tenascin-C and TNfn4,5 were retained on Sepharose 4B-linked carboxyl-terminal neurocan domains, which in BIAcore binding studies yielded a KD value of 17 nM for purified tenascin-C. We conclude that a divalent cation-dependent interaction between the COOH-terminal domain of neurocan and those fibronectin type III repeats is substantially involved in the binding of neurocan to tenascin-C. PMID- 9341126 TI - Detection and characterization of Sp1 binding activity in human chondrocytes and its alterations during chondrocyte dedifferentiation. AB - We have detected DNA binding activity for a synthetic oligonucleotide containing an Sp1 consensus sequence in nuclear extracts from human chondrocytes. Changes in the levels of Sp1 oligonucleotide binding activity were examined in nuclear extracts from freshly isolated human chondrocytes, from chondrocytes that had been cultured under conditions that allowed the maintenance of a chondrocyte specific phenotype on plastic dishes coated with the hydrogel poly(2-hydroxyethyl methacrylate), and from chondrocytes induced to dedifferentiate into fibroblast like cells by passage in monolayer culture on plastic substrata. It was observed that Sp1 binding was 2-3-fold greater in nuclear extracts from dedifferentiated chondrocytes than in nuclear extracts from either freshly isolated chondrocytes or from cells cultured in suspension. The Sp1 binding activity was specific, since it was competed by unlabeled Sp1 but not by AP1 or AP2. The addition of a polyclonal antibody against Sp1 to nuclear extracts from freshly isolated chondrocytes or to extracts isolated from chondrocytes cultured in monolayer decreased the binding of Sp1 by approximately 85%. However, when the same experiment was carried out with nuclear extracts prepared from cells cultured on poly(2-hydroxyethyl methacrylate)-coated plates, only a very slight inhibition of Sp1 binding was observed. When fragments of the COL2A1 promoter containing putative Sp1 binding sites amplified by polymerase chain reaction were examined, it was found that the amounts of DNA-protein complex formed with nuclear extracts from dedifferentiated chondrocytes were 2-3-fold greater than the amounts formed with nuclear extracts from freshly isolated chondrocytes or from cells cultured in suspension. Quantitation of DNA binding activity by titration experiments demonstrated that nuclear extracts from fibroblast-like cells contained approximately 2-fold greater Sp-1 specific binding activity than nuclear extracts from chondrocytes. The direct role of Sp1 in type II collagen gene transcription was demonstrated by co-transfection experiments of COL2A1 promoter-CAT constructs in Drosophila Schneider line L2 cells that lack Sp1 homologs. This is the first demonstration of Sp1 binding activity in human chondrocytes and of differences in Sp1 DNA binding activity between differentiated and dedifferentiated chondrocytes. PMID- 9341127 TI - Novel testis-specific protein-DNA interactions activate transcription of the mouse protamine 2 gene during spermatogenesis. AB - The mouse protamines are expressed exclusively in postmeiotic male germ cells and are crucial for the compaction of chromatin during the late stages of spermatogenesis. The temporal expression of the two mouse protamines is transcriptionally regulated in the testis. Recent studies have demonstrated that ubiquitous and testis-specific proteins bind to the promoter of the mouse protamine 2 (mP2) gene. We have performed in vitro transcription and mobility shift assays to characterize the functional significance of the protein-DNA interactions within 180 base pairs upstream of the mP2 transcription start site. Deletion and mutational analyses reveal two positive regulatory sequences for mP2 transcription at positions -59/-47 and -83/-72 of the mP2 promoter. The proximal element at -59/-47 binds to a novel testis-specific protein we name protamine activating factor 1 (PAF-1). PAF-1 reaches high levels in round spermatids at the time of mP2 transcription. Deletion of the -59/-47 sequence results in about a 3 fold reduction of mP2 transcription in vitro. Although the PAF-1 binding site (PAF-responsive element, PAF-RE), contains the sequence GTCA present in the cAMP responsive element and is very similar to the estrogen-responsive element, mobility shift assays revealed that neither the cAMP-responsive element modulator nor the estrogen receptor is the protein(s) binding to PAF-RE. Competition mobility shift assays reveal that the second positive regulatory element at -83/ 72 binds a Y-box-binding protein. Using in vitro transcription assays, a 5-fold decrease in mP2 transcription is seen when both the PAF-RE and this Y-box are deleted. These data suggest that the testis-specific PAF-1 and a Y-box-binding protein are needed to activate mP2 transcription in postmeiotic male germ cells. PMID- 9341128 TI - Mass spectrometric determination of the cleavage sites in Escherichia coli dihydroorotase induced by a cysteine-specific reagent. AB - Escherichia coli dihydroorotase contains six cysteines/subunit, which are potential ligands of structural and catalytic zinc metals at protein sites of the enzyme. Specific thiol reagents modify, in nondenaturing conditions only, two of these cysteines; these two residues are thought to be ligands of structural zinc. We report here on the localization of these two cysteines on the polypeptide chain through their cyanylation by 2-nitro-5-thiocyanobenzoic acid (NTCB) and the analysis by mass spectrometry of the protein adducts. This is the first study of E. coli dihydroorotase by mass spectrometry, allowing the accurate determination of the subunit molecular weight (38,695). Treatment of dihydroorotase by NTCB induced a cleavage N-terminal to the cyanylated cysteines. The resulting fragments visualized on electrophoresis gel have been N-terminal sequenced, and their masses were determined by electrospray-ionizing mass spectrometry. This allowed the identification of cysteines 221 and 265 as the two residues cyanylated by the reagent NTCB. Results from gel filtration of dihydroorotase cyanylated on the two cysteines indicate that these residues are involved in subunit interactions leading to the active dimer. Consistent with literature data, we assume that cysteine 221 and cysteine 265, along with the neighboring cysteines 263 and 268 arranged in cluster, are potential ligands of structural zinc of E. coli dihydroorotase. PMID- 9341129 TI - Deleted in colorectal carcinoma (DCC) binds heparin via its fifth fibronectin type III domain. AB - DCC (deleted in colorectal carcinoma) is a broadly expressed cell-surface receptor. Netrin-1 was recently identified as a DCC ligand in brain, but the possibility of other DCC ligands was suggested by the finding that an anti-DCC antibody (clone AF5) neutralized netrin-1-dependent commissural axon outgrowth without blocking DCC/netrin-1 interactions. Here we have searched for alternative cell-surface DCC ligands. A DCC-Ig fusion protein bound to neural and epithelial derived cell lines, indicating that these lines express ligand(s) for DCC. The cell-surface binding activity was mediated by the loop between beta-strands F and G of the fifth fibronectin type III repeat FNIII-D5. The loop included the sequence KNRR, which resembles heparin-binding motifs in other proteins. Heparinase and heparitinase treatment of cells reduced binding of DCC-Ig, suggesting that heparan sulfate proteoglycans are cell-surface DCC ligand(s). This was further supported by heparin blocking experiments and by binding of DCC Ig to immobilized heparan sulfate. The interaction between DCC-Ig and heparan sulfate/heparin, both on the surface of cells and immobilized on plastic, was blocked by the same anti-DCC antibody that blocks netrin-1-dependent commissural axon outgrowth. Taken together, these findings suggest that the DCC-Ig/heparin interaction may contribute to the biological activity of DCC. PMID- 9341130 TI - An antagonist for the leukemia inhibitory factor receptor inhibits leukemia inhibitory factor, cardiotrophin-1, ciliary neurotrophic factor, and oncostatin M. AB - The leukemia inhibitory factor receptor (LIF-R) is activated not only by LIF, but also by cardiotrophin-1, ciliary neurotrophic factor with its receptor, and oncostatin M (OSM). Each of these cytokines induces the hetero-oligomerization of LIF-R with gp130, a signal-transducing subunit shared with interleukin-6 and interleukin-11. The introduction of mutations into human LIF that reduced the affinity for gp130 while retaining affinity for LIF-R has generated antagonists for LIF. In the current study, a LIF antagonist that was free of detectable agonistic activity was tested for antagonism against the family of LIF-R ligands. On cells that express LIF-R and gp130, all LIF-R ligands were antagonized. On cells that also express OSM receptor, OSM was not antagonized, demonstrating that the antagonist is specific for LIF-R. Ligand-triggered tyrosine phosphorylation of both LIF-R and gp130 was blocked by the antagonist. The antagonist is therefore likely to work by preventing receptor oligomerization. PMID- 9341131 TI - FLICE is predominantly expressed as two functionally active isoforms, caspase-8/a and caspase-8/b. AB - Induction of apoptosis by the cell surface receptor CD95 (APO-1/Fas) has been shown to involve activation of a family of cysteine proteases (caspases). Recently, a new member of this family has been identified, designated FLICE (caspase-8/MACH/Mch5). FLICE is part of the CD95 death-inducing signaling complex and is therefore the most upstream caspase in the CD95 apoptotic pathway. A total of eight different isoforms of FLICE (caspase-8/a-h) have been described. To determine which isoforms are expressed in different cells we have generated a panel of monoclonal antibodies directed against all functional domains of FLICE. Using these antibodies we could show that only two of the FLICE isoforms (caspase 8/a and caspase-8/b) were predominantly expressed in cells of different origin. Both isoforms were recruited to the CD95 death-inducing signaling complex and were activated upon CD95 stimulation with similar kinetics. Taken together, only two of the eight published caspase-8 isoforms could be detected in significant amounts at the protein level. PMID- 9341132 TI - Dimerization of the delta opioid receptor: implication for a role in receptor internalization. AB - Dimerization of G-protein-coupled receptors has been increasingly noted in the regulation of their biological activity. However, its involvement in agonist induced receptor internalization is not well understood. In this study, we examined the ability of mouse delta-opioid receptors to dimerize and the role of receptor dimerization in agonist-induced internalization. Using differentially (Flag and c-Myc) epitope-tagged receptors we show that delta-opioid receptors exist as dimers. The level of dimerization is agonist dependent. Increasing concentrations of agonists reduce the levels of dimer with a corresponding increase in the levels of monomer. Interestingly, morphine does not affect the levels of either form. It has been shown that morphine, unlike other opioid agonists, does not induce receptor internalization. This suggests a relationship between the ability of agonists to reduce the levels of dimer and to induce receptor internalization. The time course of the agonist-induced decrease of delta-opioid receptor dimers is shorter than the time course of internalization, suggesting that monomerization precedes the agonist-induced internalization of the receptor. Furthermore, we found that a mutant delta-opioid receptor, with a 15-residue C-terminal deletion, does not exhibit dimerization. This mutant receptor has been shown to lack the ability to undergo agonist-induced internalization. These results suggest that the interconversion between the dimeric and monomeric forms plays a role in opioid receptor internalization. PMID- 9341133 TI - Dantrolene inhibition of sarcoplasmic reticulum Ca2+ release by direct and specific action at skeletal muscle ryanodine receptors. AB - The skeletal muscle relaxant dantrolene inhibits the release of Ca2+ from the sarcoplasmic reticulum during excitation-contraction coupling and suppresses the uncontrolled Ca2+ release that underlies the skeletal muscle pharmacogenetic disorder malignant hyperthermia; however, the molecular mechanism by which dantrolene selectively affects skeletal muscle Ca2+ regulation remains to be defined. Here we provide evidence of a high-affinity, monophasic inhibition by dantrolene of ryanodine receptor Ca2+ channel function in isolated sarcoplasmic reticulum vesicles prepared from malignant hyperthermia-susceptible and normal pig skeletal muscle. In media simulating resting myoplasm, dantrolene increased the half-time for 45Ca2+ release from both malignant hyperthermia and normal vesicles approximately 3.5-fold and inhibited sarcoplasmic reticulum vesicle [3H]ryanodine binding (Ki approximately 150 nM for both malignant hyperthermia and normal). Inhibition of vesicle [3H]ryanodine binding by dantrolene was associated with a decrease in the extent of activation by both calmodulin and Ca2+. Dantrolene also inhibited [3H]ryanodine binding to purified skeletal muscle ryanodine receptor protein reconstituted into liposomes. In contrast, cardiac sarcoplasmic reticulum vesicle 45Ca2+ release and [3H]ryanodine binding were unaffected by dantrolene. Together, these results demonstrate selective effects of dantrolene on skeletal muscle ryanodine receptors that are consistent with the actions of dantrolene in vivo and suggest a mechanism of action in which dantrolene may act directly at the skeletal muscle ryanodine receptor complex to limit its activation by calmodulin and Ca2+. The potential implications of these results for understanding how dantrolene and malignant hyperthermia mutations may affect the voltage-dependent activation of Ca2+ release in intact skeletal muscle are discussed. PMID- 9341134 TI - Metabolic impact of adenovirus-mediated overexpression of the glucose-6 phosphatase catalytic subunit in hepatocytes. AB - Glucose-6-phosphatase (G6Pase) catalyzes the hydrolysis of glucose 6-phosphate (Glu-6-P) to free glucose and, as the last step in gluconeogenesis and glycogenolysis in liver, is thought to play an important role in glucose homeostasis. G6Pase activity appears to be conferred by a set of proteins localized to the endoplasmic reticulum, including a glucose-6-phosphate translocase, a G6Pase phosphohydrolase or catalytic subunit, and glucose and inorganic phosphate transporters in the endoplasmic reticulum membrane. In the current study, we used a recombinant adenovirus containing the cDNA encoding the G6Pase catalytic subunit (AdCMV-G6Pase) to evaluate the metabolic impact of overexpression of the enzyme in primary hepatocytes. We found that AdCMV-G6Pase treated liver cells contain significantly less glycogen and Glu-6-P, but unchanged UDP-glucose levels, relative to control cells. Further, the glycogen synthase activity state was closely correlated with Glu-6-P levels over a wide range of glucose concentrations in both G6Pase-overexpressing and control cells. The reduction in glycogen synthesis in AdCMV-G6Pase-treated hepatocytes is therefore not a function of decreased substrate availability but rather occurs because of the regulatory effects of Glu-6-P on glycogen synthase activity. We also found that AdCMV-G6Pase-treated-cells had significantly lower rates of lactate production and [3-3H]glucose usage, coupled with enhanced rates of gluconeogenesis and Glu-6-P hydrolysis. We conclude that overexpression of the G6Pase catalytic subunit alone is sufficient to activate flux through the G6Pase system in liver cells. Further, hepatocytes treated with AdCMV-G6Pase exhibit a metabolic profile resembling that of liver cells from patients or animals with non-insulin-dependent diabetes mellitus, suggesting that dysregulation of the catalytic subunit of G6Pase could contribute to the etiology of the disease. PMID- 9341135 TI - A matrix form of fibronectin mediates enhanced binding of Streptococcus pyogenes to host tissue. AB - The pathogenic Gram-positive bacterium Streptococcus pyogenes (group A streptococcus) binds to fibronectin via protein F. In this study, we have investigated the binding properties of protein F to various multimeric tissue forms of fibronectin that appear on cell surfaces and in the extracellular matrix. We show that binding of S. pyogenes through protein F is more efficient to an in vitro-derived polymerized form of fibronectin (superfibronectin) than to soluble fibronectin immobilized in a solid phase. In addition, Chinese hamster ovary cells overexpressing the alpha5beta1 integrin produced an increased amount of a fibronectin matrix and consequently bound a higher number of S. pyogenes cells. Inhibition and direct binding assays using purified proteins demonstrated that binding to a fibronectin matrix involved both domains of protein F (UR and RD2) that have previously been implicated in interactions with fibronectin. Using intact S. pyogenes bacteria in which various domains of protein F were expressed as hybrids with the surface-exposed region of an unrelated protein, we revealed that, in contrast to the predominantly UR-mediated binding to soluble fibronectin, the maximal binding to the fibronectin matrix required RD2 in addition to UR. Since in some infections S. pyogenes may initially encounter a matrix form of fibronectin, these results suggest that UR and RD2 may be important for the initiation of streptococcal infectious processes. PMID- 9341136 TI - Identification of the cysteine residues involved in redox modification of plant plastidic glucose-6-phosphate dehydrogenase. AB - The cDNA sequences encoding cytosolic and light-modulated plastidic glucose-6 phosphate dehydrogenase (G6PDH) from potato were modified by polymerase chain reaction and subsequently overexpressed in Escherichia coli. Characterization of the recombinant enzymes showed that they closely resembled their native counterparts. Treatment with reduced dithiothreitol or glutathione led to inactivation of plastidic G6PDH, whereas the activity of the cytosolic isoenzyme was not influenced by reduction. As for the native enzyme, inactivation of recombinant plastidic G6PDH was accelerated by thioredoxin m and could be fully reversed by subsequent addition of oxidant. To identify the residues which are involved in redox regulation of plastidic G6PDH, each of the six cysteines in the mature protein sequence was exchanged separately for serine by site-directed mutagenesis. Two mutant proteins exhibited characteristics of the reduced wild type enzyme. Exchange of either Cys149 or Cys157 to serine abolished the regulatory properties, suggesting that these cysteine residues are the sites responsible for redox-mediated inactivation of plastidic G6PDH. PMID- 9341137 TI - Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. AB - Rab GTPases have been implicated in intracellular vesicle trafficking. Using the yeast two-hybrid screen, we have isolated a rat clone that interacts with Rab3A as well as with Rab1. The gene encodes a 20.6-kDa protein with two extensive hydrophobic domains and is broadly expressed in all tissues. This protein binds to prenylated Rab GTPases but not to other small Ras-like GTPases such as the Rho/Rac family. This prenylated Rab acceptor (PRA1) also binds specifically to the synaptic vesicle protein VAMP2 (or synaptobrevin II) but shows no affinity for VAMP1 or cellubrevin in both the yeast two-hybrid system and in vitro binding assays. This specificity resides, in part, in the proline-rich domain of VAMP2 as a chimera containing this domain of VAMP2 fused to VAMP1 is able to bind to PRA1. The transmembrane domain of VAMP2 is also essential as its deletion abolished binding to PRA1. Replacement of the deleted VAMP2 transmembrane domain by a CAAX prenylation signal can not restore binding to PRA1. This interaction is therefore distinct from that required for VAMP2 binding to either syntaxin or both syntaxin and SNAP-25. Deletion analysis on PRA1 indicates that the critical Rab- and VAMP2 interacting residues reside in two regions: the amino-terminal residues 30-54 and the extreme carboxyl-terminal domain. This dual Rab and VAMP2 binding characteristic suggests that PRA1 may serve to link these two protein families in the control of vesicle docking and fusion. PMID- 9341138 TI - Conditions for nucleotide-dependent GroES-GroEL interactions. GroEL14(groES7)2 is favored by an asymmetric distribution of nucleotides. AB - A still unresolved question regarding the mechanism of chaperonin-assisted protein folding involves the stoichiometry of the GroEL-GroES complex. This is important, because the activities of the Escherichia coli chaperonin GroEL are modulated by the cochaperonin GroES. In this report, the binding of GroES to highly purified GroEL in the presence of ATP, ADP, and the nonhydrolyzable ATP analogue, 5'-adenylyl beta,gamma-imidodiphosphate (AMP-PNP), was investigated by using the fluorescence anisotropy of succinimidyl-1-pyrenebutyrate-labeled GroES. In the presence of Mg2+-ATP and high [KCl] (10 mM), two GroES7 rings bind per one GroEL14. In contrast, in the presence of ADP or AMP-PNP only one molecule of oligomeric GroES can be tightly bound by GroEL. With AMP-PNP, binding of a small amount (<20%) of a second GroES can be detected. In the presence of ADP alone, a second GroES ring can bind to GroEL weakly and with negative cooperativity. Strikingly, addition of AMP-PNP to the solution containing preformed GroEL14(GroES7) complexes formed in the presence of ADP results in an increase in the fluorescence anisotropy. Analysis of this effect indicates that 2 mol of GroES oligomer can be bound in the presence of mixed nucleotides. A similar conclusion follows from studies in which ADP is added to an GroEL14 (GroES7) complex formed in the presence of AMP-PNP. This is the first demonstration of an asymmetric distribution of nucleotides bound on the 1:2 GroEL14 (GroES7)2 complex. The relation of the observed phenomena to the proposed mechanism of the GroEL function is discussed. PMID- 9341139 TI - A central role for beta-arrestins and clathrin-coated vesicle-mediated endocytosis in beta2-adrenergic receptor resensitization. Differential regulation of receptor resensitization in two distinct cell types. AB - G protein-coupled receptor (GPCR) sequestration to endosomes is proposed to be the mechanism by which G protein-coupled receptor kinase (GRK)-phosphorylated receptors are dephosphorylated and resensitized. The identification of beta arrestins as GPCR trafficking molecules suggested that beta-arrestins might represent critical determinants for GPCR resensitization. Therefore, we tested whether beta2-adrenergic receptor (beta2AR) resensitization was dependent upon beta-arrestins and an intact clathrin-coated vesicle endocytic pathway. The overexpression of either the beta-arrestin 1-V53D dominant negative inhibitor of beta2AR sequestration or dynamin I-K44A to block clathrin-coated vesicle-mediated endocytosis impaired both beta2AR dephosphorylation and resensitization. In contrast, resensitization of a sequestration-impaired beta2AR mutant (Y326A) was reestablished following the overexpression of either GRK2 or beta-arrestin 1. Moreover, beta2ARs did not resensitize in COS-7 cells as the consequence of impaired sequestration and dephosphorylation. However, beta2AR resensitization was restored in these cells following the overexpression of beta-arrestin 2. These findings demonstrate, using both loss and gain of function paradigms, that beta2AR dephosphorylation and resensitization are dependent upon an intact sequestration pathway. These studies also indicate that beta-arrestins play an integral role in regulating not only the desensitization and intracellular trafficking of GPCRs but their ability to resensitize. beta-Arrestin expression levels appear to underlie cell type-specific differences in the regulation of GPCR resensitization. PMID- 9341141 TI - The glycosylation of rat intestinal Muc2 mucin varies between rat strains and the small and large intestine. A study of O-linked oligosaccharides by a mass spectrometric approach. AB - The large glycosylated domains obtained from the rat intestinal mucin Muc2 were isolated from the large and small intestine of the inbred rat strains GOT-W and GOT-BW. The expression of the rat Muc2 in the large intestine was confirmed immunochemically and by Northern blotting. Released oligosaccharides were structurally characterized by gas chromatography-mass spectrometry (neutral and sialylated species) or by tandem mass spectrometry (sulfated species), and a total of 63 structures was assigned. The large intestinal oligosaccharides were found to be identical between the strains, while the small intestinal glycosylation differed. Until now, detailed structural analysis of oligosaccharides isolated from a single mucin core or mucin domain with different origin have not been performed, and the information of different mucin glycoforms has been limited to immunochemistry. Blood group A-determinants (GalNAcalpha1 3(Fucalpha1-2)Galbeta1-, and structures related to the blood group Sda/Cad related epitope NeuAc/NeuGcalpha1-3(GalNAcbeta1-4)Galbeta1-, were found in GOT-BW small intestine, and also in both large intestines. Blood group H-determinants and NeuAc/NeuGcalpha1-3Galbeta1- were found in all samples. Core 1 (Galbeta1 3GalNAcalpha1-), core 2 (Galbeta1-3(GlcNAcbeta1-6)GalNAcalpha1-), core 3 (GlcNAcbeta1-3GalNAcalpha1-), and core 4 (GlcNAcbeta1-3(GlcNAcbeta1 6)GalNAcalpha1- were also found in all the samples. The large intestine were enriched in sulfated oligosaccharides and the small intestine contained higher amounts of sialylated species. Sulfation were found exclusively on C-6 of GlcNAc. PMID- 9341140 TI - Gastrin and phorbol 12-myristate 13-acetate regulate the human histidine decarboxylase promoter through Raf-dependent activation of extracellular signal regulated kinase-related signaling pathways in gastric cancer cells. AB - Gastrin stimulates transcription of the human histidine decarboxylase (HDC) gene through binding to the G-protein-coupled cholecystokinin-B/gastrin receptor. We have explored the possibility that mitogen-activated protein kinase cascades play a role in mediating the effects of gastrin on transcription in a gastric cancer (AGS-B) cell line. Gastrin and phorbol 12-myristate 13-acetate (PMA) treatment of AGS-B cells was found to increase the phosphorylation of tyrosine residues of extracellular signal-regulated kinases (ERKs) 1 and 2 and increase ERK activity as determined by the in vitro phosphorylation of myelin basic protein. Reporter gene assays also demonstrated that gastrin and PMA stimulated Elk-1- and c-Myc dependent transactivation, consistent with gastrin- and PMA-induced activation of ERKs. Overexpression of wild type ERK-1 and ERK-2 or activation of endogenous ERKs using activated MEK-1 (mitogen-activated protein kinase kinase or ERK kinase) overexpression stimulated HDC promoter activity in a dose-dependent fashion. Interruption of the ERK-related pathway using expression vectors for kinase-deficient ERKs or an ERK-specific phosphatase (PAC-1) blocked gastrin- and PMA-stimulated HDC promoter activity. In contrast, inhibition of the Jun kinase pathway using an interfering dominant negative SEK-1 (stress-activated protein kinase/ERK-1) mutant did not inhibit HDC promoter activity. Furthermore, whereas gastrin stimulated phosphorylation of Shc proteins and association with Grb2, activation of the HDC promoter was not influenced by expression of dominant negative Ras (N15 or N17) proteins. However, gastrin stimulated Raf-1 kinase activity, and activation of the HDC promoter was blocked by coexpression of a dominant negative Raf-1 construct. Overall, these data demonstrate that gastrin regulates HDC transcription in a Rafdependent, Ras-independent fashion predominantly through activation of the ERK-related pathway. PMID- 9341142 TI - Adhesion and activation of human platelets induced by convulxin involve glycoprotein VI and integrin alpha2beta1. AB - We analyzed the interaction of convulxin (Cvx), a 72-kDa protein isolated from the venom of Crotalus durissus terrificus, with human platelets. Cvx is a potent platelet agonist that induces an increase in the intracellular Ca2+ concentration ([Ca2+]i), granule exocytosis and aggregation. 125I-Labeled Cvx binds specifically and rapidly to platelets at binding sites of high and moderate affinity. Platelets adhere to immobilized Cvx in a time-dependent but cation independent manner. Platelet exocytosis and aggregation induced by Cvx were inhibited by an anti-integrin alpha2beta1 monoclonal antibody (6F1) and by the Fab fragments of a polyclonal anti-glycoprotein VI (GPVI) antibody. Both the adhesion of platelets to Cvx and the Cvx-induced increase in [Ca2+]i were inhibited by anti-GPVI Fab fragments but not by 6F1. Ligand blotting assay showed that 125I-Cvx binds to a 57-kDa platelet protein with an electrophoretic mobility identical to that of GPVI. In addition, we observed the following: (i) 125I-Cvx binds to GPVI immunoprecipitated by the anti-GPVI antibody from a platelet lysate, and (ii) Cvx inhibits the binding of anti-GPVI IgG to GPVI. Taken together, these results demonstrate that GPVI behaves as a Cvx receptor and that the alpha2beta1 integrin appears to be involved in the later stages of Cvx induced platelet activation, i.e. exocytosis and aggregation. PMID- 9341143 TI - Structure of cDNAs encoding human eukaryotic initiation factor 3 subunits. Possible roles in RNA binding and macromolecular assembly. AB - The mammalian translation initiation factor 3 (eIF3), is a multiprotein complex of approximately 600 kDa that binds to the 40 S ribosome and promotes the binding of methionyl-tRNAi and mRNA. cDNAs encoding 5 of the 10 subunits, namely eIF3 p170, -p116, -p110, -p48, and -p36, have been isolated previously. Here we report the cloning and characterization of human cDNAs encoding the major RNA binding subunit, eIF3-p66, and two additional subunits, eIF3-p47 and eIF3-p40. Each of these proteins is present in immunoprecipitates formed with affinity-purified anti-eIF3-p170 antibodies. Human eIF3-p66 shares 64% sequence identity with a hypothetical Caenorhabditis elegans protein, presumably the p66 homolog. Deletion analyses of recombinant derivatives of eIF3-p66 show that the RNA-binding domain lies within an N-terminal 71-amino acid region rich in lysine and arginine. The N terminal regions of human eIF3-p40 and eIF3-p47 are related to each other and to 17 other eukaryotic proteins, including murine Mov-34, a subunit of the 26 S proteasome. Phylogenetic analyses of the 19 related protein sequences, called the Mov-34 family, distinguish five major subgroups, where eIF3-p40, eIF3-p47, and Mov-34 are each found in a different subgroup. The subunit composition of eIF3 appears to be highly conserved in Drosophila melanogaster, C. elegans, and Arabidopsis thaliana, whereas only 5 homologs of the 10 subunits of mammalian eIF3 are encoded in S. cerevisiae. PMID- 9341144 TI - Regulation of single chain urokinase binding, internalization, and degradation by a plasminogen activator inhibitor 1-derived peptide. AB - The internalization and degradation of cell-associated urokinase type plasminogen activator (uPA) through the alpha2-macroglobulin receptor/low density lipoprotein related receptor (alpha2MR/LRP) represent important steps in the control of plasmin formation. Complexes between two chain urokinase (tcuPA) and plasminogen activator type 1 are degraded rapidly whereas single chain urokinase (scuPA) is not, suggesting that alpha2MR/LRP requires specific epitopes in the serpin for effective function. We report an alternative mechanism that may contribute to this process. The binding of scuPA to LM-TK- cells that lack the uPA receptor was stimulated by the hexapeptide EEIIMD, corresponding to amino acids 350-355 of plasminogen activator type 1, which contacts the sequence RHRGGS, corresponding to amino acids 179-184 in uPA. EEIIMD increased the Bmax of scuPA binding 4-fold with the half-maximal effect achieved at a peptide concentration of 50 microM. Stimulation was dependent on the charge on the COOH-terminal amino acid but not on the NH2 terminus of the peptide. EEIIMD also stimulated the internalization and degradation of scuPA. Both the binding and internalization of scuPA in the presence of EEIIMD were blocked by recombinant, 39-kDa alpha2MR/LRP-associated protein as well as by an anti-alpha2MR/LRP antibody. EEIIMD also stimulated the binding of scuPA to purified alpha2MR/LRP. EEIIMD had no effect on the binding of tcuPA or of complexes between scuPA and its receptor. These results suggest that EEIIMD regulates the binding of scuPA with alpha2MR/LRP. These findings also suggest a potential mechanism by which scuPA can be cleared which is independent of activation by plasmin or binding to uPA receptor. PMID- 9341145 TI - Detailed studies on substrate structure requirements of glycoamidases A and F. AB - Glycoamidases (peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase, EC 3.5.1.52; also known as peptide: N-glycanases (PNGases) release N-linked oligosaccharides from glycopeptides and/or glycoproteins by hydrolyzing the glycosylated beta-amide bond of the asparagine side chain. The most widely used glycoamidases are those from Flavobacterium meningosepticum (glycoamidase F or PNGase F) and almond emulsin (glycoamidase A or PNGase A). To study the substrate structure requirement of these enzymes systematically, we synthesized >30 glycopeptides containing cellobiose, lactose, GlcNAc, and di-N,N' acetylchitobiose (CTB). The length of the peptide was varied from one to five amino acids, and glycosylamines were linked to either Asn or Gln located at different positions in the peptide, including NH2 and COOH termini. Neither enzyme could cleave cellobiose and lactose glycopeptides, indicating that the 2 acetamido group on the Asn-linked GlcNAc is important in the recognition by both glycoamidases A and F. GlcNAc peptides could be cleaved by both enzymes, albeit not as effectively as CTB glycopeptides. Neither enzyme requires the Asn-Xaa (Ser/Thr) sequence (required for N-glycosylation) for activity. Glycoamidase A could even hydrolyze a Gln-bound CTB glycopeptide, whereas the action of glycoamidase F on this substrate is minimal. While glycoamidase A could act on CTB dipeptides, glycoamidase F preferred a tripeptide or longer. The Km and Vmax values of glycoamidase A for t-butoxycarbonyl-(CTB)-Asn-Ala-Ser-OMe were 2.1 mM and 0.66 micromol/min/mg, respectively. A natural glycodipeptide, Man9-GlcNAc2 Asn-Phe, was also completely hydrolyzed by glycoamidase A. PMID- 9341146 TI - The role of glycine 99 in L-lactate monooxygenase from Mycobacterium smegmatis. AB - Glycine 99 in L-lactate monooxygenase (LMO) from Mycobacterium smegmatis was mutated to serine and threonine, and the resultant mutants were studied extensively to explore the role of this residue in maintaining monooxygenase activity and in controlling the reactivity with molecular oxygen. Both mutants were observed to lose monooxygenase activity completely and generate H2O2 and pyruvate as reaction products. However, the mutants have much lower activities than a true L-lactate oxidase. The oxygen reactivities of the reduced and semiquinone forms of the mutant enzymes were significantly different from those of wild type enzyme. These results confirm our previous suggestion that the electronic interactions in the active site are a crucial factor that governs the oxygen reactivity of the enzyme (Sun, W., Williams, C. H., Jr., and Massey, V. (1996) J. Biol. Chem. 271, 17226-17233). In addition, the mutants cause a dramatic decrease of the rate of flavin reduction by L-lactate compared with the wild type enzyme, mainly due to the much lower stabilization of the transition state. PMID- 9341147 TI - Aspartate 171 is the major primate-specific determinant of human growth hormone. Engineering porcine growth hormone to activate the human receptor. AB - It has been known for more than 4 decades that only primate growth hormones are effective in primate species, but it is only with the availability of the 2.8 A structure of the human growth hormone (hGH).hGH-binding protein (hGHBP)2 complex that Souza and co-workers (Souza, S. C., Frick, G. P., Wang, X., Kopchick, J. J., Lobo, R. B., and Goodman, H. M. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 959 963) were able to provide evidence that Arg-43 on the primate receptor is responsible. Here we have examined systematically the interaction between Arg-43 (primate receptor) or Leu-43 (non-primate receptors) and their complementary hormone residues Asp-171 (primate GH) and His-170 (non-primate hormones) in a four-way comparison involving exchanges of histidine and aspartate and exchanges of arginine and leucine. BAF/B03 lines were created and characterized which stably expressed hGH receptor, R43L hGH receptor, rabbit GH receptor, and L43R rabbit GH receptor. These were examined for site 1 affinity, for the ability to bind intact cells, and for proliferative biopotency using hGH, D171H hGH, porcine GH, or H170D porcine GH. We find that the single interaction between Arg-43 and His-170/171 is sufficient to explain virtually all of the primate species specificity, and this is congruent with the crystal structure. Accordingly, for the first time we have been able to engineer a non-primate hormone to bind to and activate the human GH receptor. PMID- 9341148 TI - Structure-function analysis of the diphtheria toxin receptor toxin binding site by site-directed mutagenesis. AB - Diphtheria toxin (DT) binds to the epidermal growth factor (EGF)-like domain of human membrane-anchored heparin-binding EGF-like growth factor (proHB-EGF), the human DT receptor (DTR). DT does not bind to mouse proHB-EGF because of amino acid substitutions within the EGF-like domain. We made 10 independent mutants, replacing a single amino acid within the EGF-like domain of human DTR/proHB-EGF with the corresponding amino acid residue in mouse proHB-EGF. The mutant proteins were transiently expressed in mouse L cells either expressing or not expressing DRAP27/CD9, and DT binding was measured. DT binding activity of GST fusion proteins containing the mutated EGF-like domain was also determined by a cell free binding assay. The largest effect was seen with E141H, and second largest effects were seen with F115Y and L127F in all of the assay systems. We conclude that Phe115, Leu127, and Glu141 are critical amino acid residues for DT binding. A computer model of the tertiary structure of the EGF-like domain of human DTR/proHB-EGF was made. The model predicts that three amino acid residues critical for DT binding activity, Phe115, Leu127, and Glu141, are all located on the same face of the EGF-like domain, suggesting that this face of DTR/proHB-EGF interacts with the receptor-binding domain of DT. PMID- 9341149 TI - Diazaborine resistance in the yeast Saccharomyces cerevisiae reveals a link between YAP1 and the pleiotropic drug resistance genes PDR1 and PDR3. AB - We have investigated the mechanisms underlying resistance to the drug diazaborine in Saccharomyces cerevisiae. We used UV mutagenesis to generate resistant mutants, which were divided into three different complementation groups. The resistant phenotype in these groups was found to be caused by allelic forms of the genes AFG2, PDR1, and PDR3. The AFG2 gene encodes an AAA (ATPases associated to a variety of cellular activities) protein of unknown function, while PDR1 and PDR3 encode two transcriptional regulatory proteins involved in pleiotropic drug resistance development. The isolated PDR1-12 and PDR3-33 alleles carry mutations that lead to a L1044Q and a Y276H exchange, respectively. In addition, we report that overexpression of Yap1p, the yeast homologue of the transcription factor AP1, results in a diazaborine-resistant phenotype. The YAP1-mediated diazaborine resistance is dependent on the presence of functional PDR1 and PDR3 genes, although PDR3 had a more pronounced effect. These results provide the first evidence for a functional link between the Yap1p-dependent stress response pathway and Pdr1p/Pdr3p-dependent development of pleiotropic drug resistance. PMID- 9341150 TI - Efficient transfer of synthetic ribozymes into cells using hemagglutinating virus of Japan (HVJ)-cationic liposomes. Application for ribozymes that target human t cell leukemia virus type I tax/rex mRNA. AB - We investigated the usefulness of ribozymes in inhibiting the expression of human T-cell leukemia virus type I (HTLV-I) gene. Two hammerhead ribozymes that were against HTLV-I rex (RR) and tax (TR) mRNA were synthesized. Both ribozymes were sequence-specific in the in vitro cleavage analysis of run-off transcripts from tax/rex cDNA. Intracellular activities of the ribozymes were studied in HTLV-I tax cDNA-transfected rat embryonic fibroblasts (Rat/Tax cells), which expressed the Tax but not Rex. Ribozymes were delivered into cells using anionic or cationic liposomes fused with hemagglutinating virus of Japan (HVJ). Cellular uptake of ribozymes complexed with HVJ-cationic liposomes was 15-20 times higher cellular uptake than naked ribozymes, and 4-5 times higher than that of ribozymes complexed with HVJ-anionic liposomes. HVJ-cationic liposomes promoted accumulation of ribozymes in cytoplasm and accelerated transport to the nucleus. Tax protein levels were decreased about 95% and were five times lower when the same amount of TR was introduced into the cells using HVJ-cationic, rather than HVJ-anionic liposomes. Inactive ribozyme and tax antisense oligodeoxynucleotides reduced Tax expression by about 20%, whereas RR and tax sense oligodeoxynucleotides had no effect. These results suggest that the ribozymes' effect against tax mRNA was sequence-specific, and HVJ-cationic liposomes can be useful for intracellular introduction of ribozymes. PMID- 9341152 TI - Biosynthesis, distinct post-translational modifications, and functional characterization of lymphoma proprotein convertase. AB - Proprotein convertases are responsible for the endoproteolytic processing of prohormones, neuropeptide precursors, and other proproteins within the constitutive and regulated secretory pathways. Cleavage occurs carboxyl terminally of basic amino acid motifs, such as RX(K/R)R, RXXR, and (R/K)R. As already available for the other known mammalian members of this enzyme family, we here define structural and functional features of human lymphoma proprotein convertase (LPC). Analysis of expression of recombinant LPC in stably transfected Chinese hamster ovary cells reveals biosynthesis of a 92-kDa nonglycosylated precursor (proLPC) and a 102-kDa endoglycosidase H-sensitive glycosylated form of proLPC. Only the latter is further processed and after propeptide removal converted into a complexly N-glycosylated mature form of LPC of about 92 kDa. Co expression experiments of truncated LPC with an active site mutant of LPC (LPCS265A) indicate that prodomain removal of LPC occurs via an autoproteolytic, intramolecular mechanism, as was demonstrated before for some of the other members of this enzyme family. Prodomain removal is shown to be required for LPC to exit the endoplasmic reticulum. As far as subcellular localization is concerned, immunocytochemical, ultrastructural, and biochemical analyses show that LPC is concentrated in the trans-Golgi network, associated with membranes, and not secreted. Carboxyl-terminal domains are critically involved in this cellular retention, because removal of both the hydrophobic region and the cytoplasmic tail of LPC results in secretion. Of interest are the observations that LPC is not phosphorylated like furin but is palmitoylated in its cytoplasmic tail. Finally, substrate specificity of LPC is similar to that of furin but not identical. Whereas for furin a basic substrate residue at position P-2 is dispensable, it is essential for LPC. For optimal LPC substrate processing activity, an arginine at position P-6 is preferred over an arginine at P-4. PMID- 9341153 TI - Delta and kappa opioid receptors are differentially regulated by dynamin dependent endocytosis when activated by the same alkaloid agonist. AB - Many alkaloid drugs used as analgesics activate multiple opioid receptors. Mechanisms that distinguish the actions of these drugs on the regulation of individual micro, delta, and kappa receptors are not understood. We have observed that individual cloned opioid receptors differ significantly in their regulation by rapid endocytosis in the presence of alkaloid drug etorphine, a potent agonist of mu, delta, and kappa opioid receptors. Internalization of epitope-tagged delta opioid receptors from the plasma membrane is detectable within 10 min in the presence of etorphine. In contrast, kappa receptors expressed in the same cells remain in the plasma membrane and are not internalized for >/=60 min, even when cells are exposed to saturating concentrations of etorphine. The rapid internalization of delta receptors is specifically inhibited in cells expressing K44E mutant dynamin I, suggesting that type-specific internalization of opioid receptors is mediated by clathrin-coated pits. Examination of a series of chimeric mutant kappa/delta receptors indicates that at least two receptor domains, including the highly divergent carboxyl-terminal cytoplasmic tail, determine the type specificity of this endocytic mechanism. We conclude that structurally homologous opioid receptors are differentially sorted by clathrin mediated endocytosis following activation by the same agonist ligand. These studies identify a fundamental mechanism of receptor regulation mediating type specific effects of analgesic drugs that activate more than one type of opioid receptor. PMID- 9341151 TI - Purification and characterization of the human SR 31747A-binding protein. A nuclear membrane protein related to yeast sterol isomerase. AB - SR 31747A, defined as a sigma ligand, is a novel immunosuppressive agent that blocks proliferation of human and mouse lymphocytes. Using a radiolabeled chemical probe, we here purified a target of SR 31747A and called it SR 31747A binding protein (SR-BP). Purified SR-BP retained its binding properties and migrated on SDS-polyacrylamide gel as a Mr 28,000 protein. Cloning of the cDNA encoding human SR-BP shows an open reading frame for a 223-amino acid protein, which is homologous to the recently cloned sigma 1 receptor. Interestingly, the deduced amino acid sequence was found to be related to fungal C8-C7 sterol isomerase, encoded by the ERG2 gene. The ERG2 gene product has been identified recently as the molecular target of SR 31747A that mediates antiproliferative effects of the drug in yeast. Northern blot analysis of SR-BP gene expression revealed a single transcript of 2 kilobases which was widely expressed among organs, with the highest abundance in liver and the lowest abundance in brain. Subcellular localization analysis in various cells, using a specific monoclonal antibody raised against SR-BP, demonstrated that this protein was associated with the nuclear envelope. When studying the binding of SR 31747A on membranes from yeast expressing SR-BP, we found a pharmacological profile of sigma 1 receptors; binding was displaced by (+)-pentazocine, haloperidol, and (+)-SKF 10,047, with (+)-SKF 10, 047 being a more potent competitor than (-)-SKF 10,047. Scatchard plot analysis revealed Kd values of 7.1 nM and 0.15 nM for (+)-pentazocine and SR 31747A, respectively, indicating an affinity of SR-BP 50-fold higher for SR 31747A than for pentazocine. Additionally, we showed that pentazocine, a competitive inhibitor of SR 31747A binding, also prevents the immunosuppressive effect of SR 31747A. Taken together, these findings strongly suggest that SR-BP represents the molecular target for SR 31747A in mammalian tissues, which could be critical for T cell proliferation. PMID- 9341154 TI - Thermus thermophilis dnaX homolog encoding gamma- and tau-like proteins of the chromosomal replicase. AB - This report identifies the dnaX homolog from Thermus thermophilis. Replicases from bacteria to humans contain subunits that are homologous to one another. These homologs are subunits of a clamp loading apparatus that loads sliding clamps onto DNA, which in turn act as mobile tethers for the replication machinery. In Escherichia coli, two of these subunits (gamma and tau) are encoded by one gene (dnaX) in nearly equal amounts by way of an efficient translational frameshift. The gamma and tau subunits form the central touchpoint that holds together two DNA polymerases with one clamp loading apparatus to form the E. coli chromosomal replicase, DNA polymerase III holoenzyme. The E. coli holoenzyme is an efficient replication machine that simultaneously replicates both strands of duplex DNA. The T. thermophilis dnaX homolog also contains a frameshift signature and produces both tau- and gamma-like proteins. Recombinant T. thermophilis tau- and gamma-like proteins, expressed in E. coli, have an oligomeric state similar to that of their E. coli counterparts and display ATPase activity that is stimulated by DNA. These results imply that T. thermophilis utilizes a DNA polymerase III holoenzyme replication machinery similar to that of E. coli. PMID- 9341155 TI - Transport of rat liver glycine N-methyltransferase into rat liver nuclei. AB - Rat liver cytosolic glycine N-methyltransferase (GNMT) catalyzes the S adenosylmethionine-dependent methylation of glycine to sarcosine. It is comprised of four identical 292-amino acid residue subunits. Recently, evidence has been provided to show that GNMT is identical to the cytosolic receptor for benzo[a]pyrene, which induces cytochrome P450 1A1 gene expression. In the present study we show that chemical modification of purified rat liver GNMT with fluorescein isothiocyanate (FITC) resulted in dissociation of the tetrameric enzyme and was accompanied by loss of enzyme activity. Amino acid sequence analysis of the FITC-labeled peptides obtained by hydrolysis of the modified protein with Staphylococcus aureus V8 protease revealed that lysines 45, 89, 92, 96, 122, and 147 were modified. Lys-122 and Lys-147 were derivatized in tetrameric, dimeric, and monomeric forms of the enzyme. Lysines 45, 89, 92, and 96 were derivatized only in monomeric GNMT, suggesting that modification of these residues resulted in GNMT dissociation. The modified monomeric GNMT was quickly transported into isolated rat liver nuclei. This transport was specific for the GNMT monomer, since neither tetramer nor dimer was able to enter the nuclei. Bovine carbonic anhydrase, similar in size to the GNMT monomer, was labeled with FITC to a similar extent but was not transported into the nuclei. Disruption of the nuclei containing fluorescein-labeled GNMT and subsequent extraction of the nuclear lysate with both high and low salt buffers recovered FITC-GNMT only in the chromatin pellet. Our study supports the suggestion of an additional function for GNMT, probably connected with regulation of cytochrome P450 1A1 gene expression. PMID- 9341157 TI - Transforming growth factor beta peptide antagonists and their conversion to partial agonists. AB - Transforming growth factor beta (TGF-beta) has been implicated in the pathogenesis of various human diseases. Synthetic TGF-beta antagonists therefore could have therapeutic utility. Here we show the development of such compounds. Three synthetic pentacosapeptides designated beta125-(41-65), beta225-(41-65), and beta325-(41-65), whose amino acid sequences correspond to the 41st to 65th amino acid residues of TGF-beta1, TGF-beta2, and TGF-beta3, respectively, inhibit the binding of 125I-labeled TGF-beta isoforms to TGF-beta receptors in mink lung epithelial cells with IC50 of approximately 0.06-2 microM. beta125-(41-65) blocks TGF-beta1-induced growth inhibition and TGF-beta1-induced plasminogen activator inhibitor-1 expression in these cells. The variants designated beta125-(41 65)W52A/D55A and beta325-(41-65)R52A/D55A, in which both Trp52/Arg52 and Asp55 are replaced by alanine residues, do not have TGF-beta antagonist activity. Multiple conjugation of beta125-(41-65) to carrier proteins enhances its antagonist activity but also confers partial agonist activity as measured by DNA synthesis inhibition. These results suggest that the (W/R)XXD motif is important for the activities of these TGF-beta peptide antagonists and that this motif may be the active site sequence of TGF-beta. PMID- 9341156 TI - Functional characterization of the ocular prostaglandin f2alpha (PGF2alpha) receptor. Activation by the isoprostane, 12-iso-PGF2alpha. AB - Prostaglandin F2alpha (PGF2alpha) is a product of cyclooxygenase-catalyzed metabolism of arachidonic acid. Recently, PGF2alpha analogs have been hypothesized to reduce intraocular pressure via relaxation of the ciliary muscle. To investigate the molecular basis of PGF2alpha receptor (FP) activation in the eye, we cloned the FP from a human ciliary body (hcb) cDNA library. The open reading frame of the hcb-FP cDNA was identical to the uterine FP cDNA. The hcb-FP appeared to be predominantly membrane-localized, as visualized by an FP-specific peptide antibody, and coupled to inositol phosphate formation when stably expressed in HEK 293 cells. Interestingly, the hcb-FP could also be activated by the F2 isoprostane, 12-iso-PGF2alpha, in addition to its cognate ligand, PGF2alpha. 12-iso-PGF2alpha was less potent (EC50 = 5 microM) than PGF2alpha (EC50 = 10 nM) in generating inositol phosphates via the hcb-FP in HEK 293 cells. Both ligands also stimulated mitogenesis in NIH 3T3 cells. Although 12-iso PGF2alpha caused a dose-dependent activation of the FP, it failed to activate the recombinant human prostacyclin receptor and caused only minimal activation of the thromboxane receptor isoforms stably expressed in HEK 293 cells. Four additional F2 isoprostanes, 8-iso-PGF2alpha, IPF2alpha-I, IPF2alpha-III, and 9beta,11beta PGF2, caused trivial, or no, activation of the FP. Consistent with these observations, only PGF2alpha and 12-iso-PGF2alpha caused rapid homologous desensitization of FP and also exhibited cross-desensitization, with PGF2alpha resulting in a maximum of approximately 60% desensitization. The human FP may thus be activated specifically, by the free radical-catalyzed F2 isoprostane, 12 iso-PGF2alpha, in addition to the cyclooxygenase product, PGF2alpha. Incidental receptor activation by isoprostanes may complement the actions of PGF2alpha in clinical syndromes where oxidant stress and augmented prostaglandin biosynthesis coincide. PMID- 9341158 TI - Functional domain mapping of the clathrin-associated adaptor medium chains mu1 and mu2. AB - The clathrin-associated adaptors AP-1 and AP-2 are heterotetrameric complexes involved in the recognition of sorting signals present within the cytosolic domain of integral membrane proteins. The medium chains of these complexes, mu1 and mu2, have been implicated in two types of interaction: assembly with the beta1 and beta2 chains of the corresponding complexes and recognition of tyrosine based sorting signals. In this study, we report the results of a structure function analysis of the mu1 and mu2 chains aimed at identifying regions of the molecules that are responsible for each of the two interactions. Analyses using the yeast two-hybrid system and proteolytic digestion experiments suggest that mu1 and mu2 have a bipartite structure, with the amino-terminal one-third (residues 1-145 of mu1 and mu2) being involved in assembly with the beta chains and the carboxyl-terminal two-thirds (residues 147-423 of mu1 and 164-435 of mu2) binding tyrosine-based sorting signals. These observations support a model in which the amino-terminal one-third of mu2 is embedded within the core of the AP-2 complex, while the carboxyl-terminal two-thirds of the protein are exposed to the medium, placing this region in a position to interact with tyrosine-based sorting signals. PMID- 9341159 TI - Kinetics of interaction of the myristoylated alanine-rich C kinase substrate, membranes, and calmodulin. AB - Membrane binding of the myristoylated alanine-rich C kinase substrate (MARCKS) requires both its myristate chain and basic "effector" region. Previous studies with a peptide corresponding to the effector region, MARCKS-(151-175), showed that the 13 basic residues interact electrostatically with acidic lipids and that the 5 hydrophobic phenylalanine residues penetrate the polar head group region of the bilayer. Here we describe the kinetics of the membrane binding of fluorescent (acrylodan-labeled) peptides measured with a stopped-flow technique. Even though the peptide penetrates the polar head group region, the association of MARCKS (151-175) with membranes is extremely rapid; association occurs with a diffusion limited association rate constant. For example, kon = 10(11) M-1 s-1 for the peptide binding to 100-nm diameter phospholipid vesicles. As expected theoretically, kon is independent of factors that affect the molar partition coefficient, such as the mole fraction of acidic lipid in the vesicle and the salt concentration. The dissociation rate constant (koff) is approximately 10 s-1 (lifetime = 0.1 s) for vesicles with 10% acidic lipid in 100 mM KCl. Ca2+ calmodulin (Ca2+.CaM) decreases markedly the lifetime of the peptide on vesicles, e.g. from 0.1 to 0.01 s in the presence of 5 micrM Ca2+.CaM. Our results suggest that Ca2+.CaM collides with the membrane-bound MARCKS-(151-175) peptide and pulls the peptide off rapidly. We discuss the biological implications of this switch mechanism, speculating that an increase in the level of Ca2+-calmodulin could rapidly release phosphatidylinositol 4, 5-bisphosphate that previous work has suggested is sequestered in lateral domains formed by MARCKS and MARCKS-(151 175). PMID- 9341160 TI - SOCS-1/JAB/SSI-1 can bind to and suppress Tec protein-tyrosine kinase. AB - Tec is the prototype of a recently emerging subfamily among nonreceptor type protein-tyrosine kinases and is known to become tyrosine-phosphorylated and activated by a wide range of cytokine stimulations in hematopoietic cells. Although Tec was recently shown to be involved in the cytokine-driven activation mechanism of c-fos transcription, it is yet obscure how Tec relays the signals from cell surface receptors to the nucleus. To identify signaling molecules acting downstream of Tec, we have looked for Tec-interacting proteins (TIPs) by using the yeast two-hybrid system. Here we report the identification and characterization of a novel protein, TIP3, which has been simultaneously identified by other groups as SOCS-1, JAB, or SSI-1. TIP3 carries one Src homology 2 domain with a sequence similarity to that of CIS. In 293 cells, TIP3 associates with Tec and suppresses its kinase activity. Interestingly, TIP3 can also down-regulate the activity of Jak2 but not that of Lyn. We propose that SOCS 1/JAB/SSI-1/TIP3 is a novel type of negative regulator to a subset of protein tyrosine kinases. PMID- 9341161 TI - Networking in the hemostatic system. Integrin alphaiibbeta3 binds prothrombin and influences its activation. AB - Prothrombin activation is a pivotal event in thrombosis and hemostasis because thrombin can mediate fibrin formation and can activate and aggregate platelets. Platelet aggregation depends upon the binding of adhesive proteins to integrin alphaIIbbeta3 on the platelet surface. In the present study, a novel interface between the blood coagulation system and platelets is demonstrated by showing that 1) prothrombin binds to alphaIIbbeta3 and 2) this interaction accelerates prothrombin activation. Prothrombin bound to purified alphaIIbbeta3 in a specific, saturable, and divalent cation-dependent manner. This interaction was inhibited by certain monoclonal antibodies to alphaIIbbeta3, by the alphaIIbbeta3 ligands fibrinogen and RGD peptides, but not by thrombin or unrelated proteins. Prothrombin also interacted with alphaIIbbeta3 on resting and stimulated platelets as demonstrated by soluble ligand binding and platelet adhesion assays. Activation of prothrombin by Factor Xa alone or Factor Xa-Va was accelerated by alphaIIbbeta3, and this enhancement was blocked by a monoclonal antibody that inhibited prothrombin binding to the receptor. Taken together, these data identify a previously unrecognized linkage between platelets and the blood coagulation system that may have a significant regulatory consequence. PMID- 9341162 TI - Pathophysiology of the MELAS 3243 transition mutation. AB - Single base substitutions of the mitochondrial genome are associated with a variety of metabolic disorders. The myopathy, encephalopathy, lactic acidosis, stroke-like episodes syndrome, most frequently associated with an A to G transition mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene, is characterized by biochemical and structural alterations of mitochondria. To investigate the pathophysiology of the mutation, we established distinct Epstein Barr virus-transformed B-cell lines for analyses that harbored 30-70% of the mutated genome. Interestingly, neither an alteration of the processing of primary transcripts nor a general impairment of individual mitochondrial protein subunit synthesis rates could be observed. Nevertheless a marked decrease of cytochrome-c oxidase activity and reduced content of mitochondrial encoded subunits in the assembled respiratory complex IV was recorded on the cell line harboring 70% mutated mtDNA. Quantitative analysis of incorporation rates of the amino acid leucine into newly synthesized mitochondrial proteins, representing the functionality of the tRNALeu(UUR) in protein biosynthesis, revealed a specific decrease of this amino acid in distinct mitochondrial translation products. This observation was supported by a variation in the proteolytic fingerprint pattern. Our results suggest that the malfunctioning mitochondrial tRNALeu(UUR) leads to an alteration of amino acid incorporation into the mitochondrially synthesized subunits of the oxidative phosphorylation system, thus altering it's structure and function. PMID- 9341163 TI - Involvement of the C-terminal domain of an ATP-binding subunit in the regulation of the ABC-type nitrate/nitrite transporter of the Cyanobacterium synechococcus sp. strain PCC 7942. AB - In Synechococcus sp. strain PCC 7942, an ATP-binding cassette transporter encoded by the genes nrtA, nrtB, nrtC, and nrtD mediates active transport of nitrate and nitrite, which is inhibited by ammonium, a preferred source of nitrogen for the cyanobacterium. One of the ATP-binding subunits of the transporter, NrtC, has a distinct C-terminal domain of 380 amino acid residues. A mutant NC2, constructed by removal of this domain using genetic engineering techniques, assimilated low concentrations of nitrate and nitrite and accumulated nitrate intracellularly, showing that the domain is not essential for the transporter activities. Assimilation of low concentrations of nitrite was only partially inhibited by ammonium in NC2 but was completely inhibited in the wild-type cells. Cells of NC2 and its derivative (nitrate reductase-less strain NC4) carrying the truncated NrtC but not the cells with the wild-type NrtC accumulated nitrate intracellularly in the presence of ammonium in medium. These findings indicated that the C-terminal domain of NrtC is involved in the ammonium-promoted inhibition of the nitrate/nitrite transporter. In the presence of ammonium, NC2 could not assimilate nitrate despite its ability to accumulate nitrate intracellularly, which suggested that reduction of intracellular nitrate by nitrate reductase is also subject to inhibition by ammonium. PMID- 9341164 TI - The DNA cleavage reaction of DNA gyrase. Comparison of stable ternary complexes formed with enoxacin and CcdB protein. AB - The potent synthetic fluoroquinolones and the natural CcdB protein encoded by the F plasmid both inhibit bacterial growth by attacking DNA gyrase and by stimulating enzyme-induced breaks in bacterial DNA. The cleavage mechanisms of these structurally diverse compounds were analyzed by purifying and characterizing stable ternary complexes of enoxacin and CcdB protein with gyrase bound to a strong gyrase binding site from bacteriophage Mu. Three differences between enoxacin- and CcdB-derived complexes were discovered. 1) Enoxacin binds to the DNA active site and alters the breakage/reunion activity of the enzyme. CcdB binds gyrase-DNA complexes but does not influence enzymatic activity directly. 2) Complexes that produce DNA cleavage with enoxacin are reversible, whereas similar complexes made with CcdB protein are not. 3) Enoxacin stimulates cleavage of both relaxed and supercoiled forms of DNA in the absence of ATP, whereas CcdB induces cleavage only after many cycles of ATP-dependent breakage and reunion. These differences in mechanisms can be explained by a model in which enoxacin induces formation of a novel "cleavable" complex, whereas CcdB protein traps a very rare "cleaved" conformation of the enzyme. PMID- 9341165 TI - The role of base flipping in damage recognition and catalysis by T4 endonuclease V. AB - The process of moving a DNA base extrahelical (base flipping) has been shown in the co-crystal structure of a UV-induced pyrimidine dimer-specific glycosylase, T4 endonuclease V, with its substrate DNA. Compared with other enzymes known to use base flipping, endonuclease V is unique in that it moves the base opposite the target site extrahelical, rather than moving the target base itself. Utilizing substrate analogs and catalytically inactive mutants of T4 endonuclease V, this study investigates the discrete steps involved in damage recognition by this DNA repair enzyme. Specifically, fluorescence spectroscopy analysis shows that fluorescence changes attributable to base flipping are specific for only the base directly opposite either abasic site analogs or the 5'-thymine of a pyrimidine dimer, and no changes are detected if the 2-aminopurine is moved opposite the 3'-thymine of the pyrimidine dimer. Interestingly, base flipping is not detectable with every specific binding event suggesting that damage recognition can be achieved without base flipping. Thus, base flipping does not add to the stability of the specific enzyme-DNA complex but rather induces a conformational change to facilitate catalysis at the appropriate target site. When used in conjunction with structural information, these types of analyses can yield detailed mechanistic models and critical amino acid residues for extrahelical base movement as a mode of damage recognition. PMID- 9341166 TI - cDNA cloning, tissue distribution, and identification of the catalytic triad of monoglyceride lipase. Evolutionary relationship to esterases, lysophospholipases, and haloperoxidases. AB - Monoglyceride lipase catalyzes the last step in the hydrolysis of stored triglycerides in the adipocyte and presumably also complements the action of lipoprotein lipase in degrading triglycerides from chylomicrons and very low density lipoproteins. Monoglyceride lipase was cloned from a mouse adipocyte cDNA library. The predicted amino acid sequence consisted of 302 amino acids, corresponding to a molecular weight of 33,218. The sequence showed no extensive homology to other known mammalian proteins, but a number of microbial proteins, including two bacterial lysophospholipases and a family of haloperoxidases, were found to be distantly related to this enzyme. By means of multiple sequence alignment and secondary structure prediction, the structural elements in monoglyceride lipase, as well as the putative catalytic triad, were identified. The residues of the proposed triad, Ser-122, in a GXSXG motif, Asp-239, and His 269, were confirmed by site-directed mutagenesis experiments. Northern blot analysis revealed that monoglyceride lipase is ubiquitously expressed among tissues, with a transcript size of about 4 kilobases. PMID- 9341167 TI - The geranylgeranyltransferase-I inhibitor GGTI-298 arrests human tumor cells in G0/G1 and induces p21(WAF1/CIP1/SDI1) in a p53-independent manner. AB - Recently we have shown that in fibroblasts (NIH 3T3 and Rat-1 cells) inhibition of protein geranylgeranylation leads to a G0/G1 arrest, whereas inhibition of protein farnesylation does not affect cell cycle distribution. Here we demonstrate that in human tumor cells the geranylgeranyltransferase-I (GGTase-I) inhibitor GGTI-298 blocked cells in G0/G1, whereas the farnesyltransferase (FTase) inhibitor FTI-277 showed a differential effect depending on the cell line. FTI-277 accumulated Calu-1 and A-549 lung carcinoma and Colo 357 pancreatic carcinoma cells in G2/M, T-24 bladder carcinoma, and HT-1080 fibrosarcoma cells in G0/G1, but had no effect on cell cycle distribution of pancreatic (Panc-1), breast (SKBr 3 and MDAMB-231), and head and neck (A-253) carcinoma cells. Furthermore, treatment of Calu-1, Panc-1, Colo 357, T-24, A-253, SKBr 3, and MDAMB-231 cells with GGTI-298, but not FTI-277, induced the protein expression levels of the cyclin-dependent kinase inhibitor p21WAF. HT-1080 and A-549 cells had a high basal level of p21WAF, and GGTI-298 did not further increase these levels. Furthermore, GGTI-298 also induces the accumulation of large amounts of p21WAF mRNA in Calu-1 cells, a cell line that lacks the tumor suppressor gene p53. There was little effect of GGTI-298 on the cellular levels of another cyclin dependent kinase inhibitor p27KIP as well as cyclin E and cyclin D1. These results demonstrate that GGTase-I inhibitors arrest cells in G0/G1 and induce accumulation of p21WAF in a p53-independent manner and that FTase inhibitors can interfere with cell cycle events by a mechanism that involves neither p21WAF nor p27KIP. The results also point to the potential of GGTase-I inhibitors as agents capable of restoring growth arrest in cells lacking functional p53. PMID- 9341168 TI - Thyroid Na+/I- symporter. Mechanism, stoichiometry, and specificity. AB - The rat thyroid Na+/I- symporter (NIS) was expressed in Xenopus laevis oocytes and characterized using electrophysiological, tracer uptake, and electron microscopic methods. NIS activity was found to be electrogenic and Na+-dependent (Na+ >> Li+ >> H+). The apparent affinity constants for Na+ and I- were 28 +/- 3 mM and 33 +/- 9 microM, respectively. Stoichiometry of Na+/anion cotransport was 2:1. NIS was capable of transporting a wide variety of anions (I-, ClO3-, SCN-, SeCN-, NO3-, Br-, BF4-, IO4-, BrO3-, but perchlorate (ClO4-) was not transported. In the absence of anion substrate, NIS exhibited a Na+-dependent leak current (approximately 35% of maximum substrate-induced current) with an apparent Na+ affinity of 74 +/- 14 mM and a Hill coefficient (n) of 1. In response to step voltage changes, NIS exhibited current transients that relaxed with a time constant of 8-14 ms. Presteady-state charge movements (integral of the current transients) versus voltage relations obey a Boltzmann relation. The voltage for half-maximal charge translocation (V0.5) was -15 +/- 3 mV, and the apparent valence of the movable charge was 1. Total charge was insensitive to [Na+]o, but V0.5 shifted to more negative potentials as [Na+]o was reduced. NIS charge movements are attributed to the conformational changes of the empty transporter within the membrane electric field. The turnover rate of NIS was >/=22 s-1 in the Na+ uniport mode and >/=36 s-1 in the Na+/I- cotransport mode. Transporter density in the plasma membrane was determined using freeze-fracture electron microscopy. Expression of NIS in oocytes led to a approximately 2. 5-fold increase in the density of plasma membrane protoplasmic face intramembrane particles. On the basis of the kinetic results, we propose an ordered simultaneous transport mechanism in which the binding of Na+ to NIS occurs first. PMID- 9341169 TI - Synaptojanin forms two separate complexes in the nerve terminal. Interactions with endophilin and amphiphysin. AB - Endophilin is a recently discovered src homology 3 domain-containing protein that is a major in vitro binding partner for synaptojanin. To further characterize endophilin, we generated an antipeptide antibody. Endophilin is enriched in the brain, and immunofluorescence analysis reveals a high concentration of the protein in synaptic terminals, where it colocalizes with synaptojanin. In vitro binding assays demonstrate that endophilin binds through its src homology 3 domain to synaptojanin, and immunoprecipitation analysis with the antiendophilin antibody reveals that endophilin is stably associated with synaptojanin in the nerve terminal. Immunoprecipitation with an antibody against amphiphysin I and II, which interact through their src homology 3 domains with dynamin and synaptojanin at sites distinct from those for endophilin, reveals a second stable complex, which includes dynamin and synaptojanin but excludes endophilin. These data demonstrate that synaptojanin is present in two separate complexes in the nerve terminal and support an important role for endophilin in the regulation of synaptojanin function. PMID- 9341170 TI - Tissue-specific regulation of mouse core 2 beta-1,6-N acetylglucosaminyltransferase. AB - Mouse kidney beta-1,6-GlcNAc-transferase (GNT) is the key enzyme for the synthesis of a glycosphingolipid (Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-6(Galbeta1 3)GalNAcbeta1-3Galalph a1-4Galbeta1-4Glcbeta1-ceramide) that contains the LeX trisaccharide epitope at its nonreducing terminus. The expression of this glycolipid in the kidney is polymorphic; it is expressed in BALB/c but not DBA/2 mice; and a single autosomal gene (Gsl5) is responsible for this polymorphism. We report here the cDNA sequence that encodes the kidney GNT of BALB/c mice, which possess a wild-type Gsl5 gene. The deduced amino acid sequence exhibits 84% identity to that of human core 2 beta-1,6-GlcNAc-transferase, which suggests that kidney GNT is a mouse homologue of human core 2 beta-1, 6-GlcNAc-transferase. The GNT mRNA is expressed abundantly in the kidney, but was not detected in other BALB/c organs or in the kidneys of DBA/2 mice by Northern blot analysis. In addition, we were able to clone and sequence another homologous cDNA from the submandibular gland. The two sequences differ only in their 5'-untranslated region. The submandibular gland type of cDNA was detected in various organs of DBA/2 mice by reverse transcription-polymerase chain reaction, which indicates that the submandibular gland type is ubiquitous and that its expression is not regulated by the Gsl5 gene. Results obtained using the long accurate polymerase chain reaction method indicate that the GNT gene is approximately 45 kilobases long, and the order of the exons from the 5'-end is exon 1 of the kidney type, exon 1 of the ubiquitous type, exon 2, and exon 3. Exons 2 and 3 are present in both transcripts, and the translated region is in exon 3. These data suggest that the expression of GNT is regulated by an alternative splicing mechanism and also probably by tissue-specific enhancers and that Gsl5 regulates the expression of GNT only in the kidney. PMID- 9341171 TI - The histone acetyltransferase activity of human GCN5 and PCAF is stabilized by coenzymes. AB - Here we report that PCAF and human GCN5, two related type A histone acetyltransferases, are unstable enzymes that under the commonly used assay conditions are rapidly and irreversibly inactivated. In addition, we report that free histone H1, although not acetylated in vivo, is a preferred and convenient in vitro substrate for the study of PCAF, human GCN5, and possibly other type A histone acetyltransferases. Using either histone H1 or histone H3 as substrates, we find that preincubation with either acetyl-CoA or CoA stabilizes the acetyltransferase activities of PCAF, human GCN5 and an enzymatically active PCAF deletion mutant containing the C-terminal half of the protein. The stabilization requires the continuous presence of coenzyme, suggesting that the acetyltransferase-coenzyme complexes are stable, while the isolated apoenzymes are not. Human GCN5 and the N-terminal deletion mutant of PCAF are stabilized equally well by preincubation with either CoA or acetyl-CoA, while intact PCAF is better stabilized by acetyl-CoA than by CoA. Intact PCAF, but not the N-terminal truncation mutant or human GCN5, is autoacetylated. These findings raise the possibility that the intracellular concentrations of the coenzymes affect the stability and therefore the nuclear activity of these acetyltransferases. PMID- 9341172 TI - Defining the minimal domain of Ku80 for interaction with Ku70. AB - The Ku protein has a critical function in the repair of double-strand DNA breaks induced for example by ionizing radiation or during VDJ recombination. Ku serves as the DNA-binding subunit of the DNA-dependent kinase and is a heterodimeric protein composed of 80- and 70-kDa subunits. We used the two-hybrid system to analyze the interaction domains of the Ku subunits and to identify possible additional partners for Ku. Screening a human cDNA library with the Ku heterodimer did not reveal any novel partners. Screening with the individual subunits, we detected only Ku70 clones interacting with Ku80 and only Ku80 clones interacting with Ku70, indicating that these are the primary partners for one another. Ku80 and Ku70 formed only heterodimers and did not homodimerize. Ku80 was restricted to interacting with just one Ku70 molecule at a time. The minimal functional interaction domain of Ku80 that interacted with Ku70 was defined. It consisted of a 28-amino acid region extending from amino acid 449 to 477. This region was crucial for interaction with Ku70, since mutation within this critical site at amino acids 453 and 454 abrogated the ability to interact with Ku70. We furthermore verified that the same region is crucial for interaction with Ku70 using in vitro co-translation of both subunits followed by an immunoprecipitation with anti-Ku70 antibodies. This interaction domain of Ku80 does not contain any motif previously recognized in protein-protein interactions. PMID- 9341173 TI - Characterization of the Rhodobacter capsulatus housekeeping RNA polymerase. In vitro transcription of photosynthesis and other genes. AB - To begin to characterize biochemically the transcriptional activation systems in photosynthetic bacteria, the Rhodobacter capsulatus RNA polymerase (RNAP) that contains the sigma70 factor (R. capsulatus RNAP/sigma70) was purified and characterized using two classical sigma70 type promoters, the bacteriophage T7A1 and the RNA I promoters. Transcription from these promoters was sensitive to rifampicin, RNase, and monoclonal antibody 2G10 (directed against the Escherichia coli sigma70 subunit). Specific transcripts were detected in vitro for R. capsulatus cytochrome c2 (cycA) and fructose-inducible (fruB) promoters and genes induced in photosynthesis (puf and puc) and bacteriochlorophyll biosynthesis (bchC). Alignment of these natural promoters activated by R. capsulatus RNAP/sigma70 indicated a preference for the sequence TTGAC at the -35 region for strong in vitro transcription. To test the -35 recognition pattern, the R. capsulatus nifA1 promoter, which exhibits only three of the five consensus nucleotides at the -35 region, was mutated to four and five of the consensus nucleotides. Although the nifA1 wild type promoter showed no transcription, the double mutated promoter exhibited high levels of in vitro transcription by the purified R. capsulatus RNAP/sigma70 enzyme. Similarities and differences between the RNAPs and the promoters of R. capsulatus and E. coli are discussed. PMID- 9341174 TI - Specificity and determinants of Sam68 RNA binding. Implications for the biological function of K homology domains. AB - Sam68, a specific target of the Src tyrosine kinase in mitosis, possesses features common to RNA-binding proteins, including a K homology (KH) domain. To elucidate its biological function, we first set out to identify RNA species that bound to Sam68 with high affinity using in vitro selection. From a degenerate 40 mer pool, 15 RNA sequences were selected that bound to Sam68 with Kd values of 12 140 nM. The highest affinity RNA sequences (Kd approximately 12-40 nM) contained a UAAA motif; mutation to UACA abolished binding to Sam68. Binding of the highest affinity ligand, G8-5, was assessed to explore the role of different regions of Sam68 in RNA binding. The KH domain alone did not bind G8-5, but a fragment containing the KH domain and a region of homology within the Sam68 subgroup of KH containing proteins was sufficient for G8-5 binding. Deletion of the KH domain or mutation of KH domain residues analogous to loss-of-function mutations in the human Fragile X syndrome gene product and the Caenorhabditis elegans tumor suppressor protein Gld-1 abolished G8-5 binding. Our results establish that a KH domain-containing protein can bind RNA with specificity and high affinity and suggest that specific RNA binding is integral to the functions of some regulatory proteins in growth and development. PMID- 9341175 TI - Serine phosphorylation-dependent association of the band 4.1-related protein tyrosine phosphatase PTPH1 with 14-3-3beta protein. AB - PTPH1 is a human protein-tyrosine phosphatase with homology to the band 4.1 superfamily of cytoskeletal-associated proteins. PTPH1 was found to associate with 14-3-3beta using a yeast two-hybrid screen, and its interaction could be reconstituted in vitro using recombinant proteins. Examination of the interaction between 14-3-3beta and various deletion mutants of PTPH1 by two-hybrid tests suggested that the integrity of the PTP is important for this binding. Although both PTPH1 and Raf-1 form complexes with 14-3-3beta, they appear to do so independently. Binding of 14-3-3beta to PTPH1 in vitro was abolished by pretreating PTPH1 with potato acid phosphatase and was greatly enhanced by pretreating with Cdc25C-associated protein kinase. Thus the association between PTPH1 and 14-3-3beta is phosphorylation-dependent. Two novel motifs RSLS359VE and RVDS853EP in PTPH1 were identified as major 14-3-3beta-binding sites, both of which are distinct from the consensus binding motif RSXSXP recently found in Raf 1. Mutation of Ser359 and Ser853 to alanine significantly reduced the association between 14-3-3beta and PTPH1. Furthermore, association of PTPH1 and 14-3-3beta was detected in several cell lines and was regulated in response to extracellular signals. These results raise the possibility that 14-3-3beta may function as an adaptor molecule in the regulation of PTPH1 and may provide a link between serine/threonine and tyrosine phosphorylation-dependent signaling pathways. PMID- 9341177 TI - Interactions between subunits of the human epithelial sodium channel. AB - The human epithelial sodium channel (hENaC) mediates Na+ transport across the apical membrane of epithelia, and mutations in hENaC result in hypertensive and salt-wasting diseases. In heterologous expression systems, maximal hENaC function requires co-expression of three homologous proteins, the alpha, beta, and gammahENaC subunits, suggesting that hENaC subunits interact to form a multimeric channel complex. Using a co-immunoprecipitation assay, we found that hENaC subunits associated tightly to form homo- and heteromeric complexes and that the association between subunits occurred early in channel biosynthesis. Deletion analysis of gammahENaC revealed that the N terminus was sufficient but not necessary for co-precipitation of alphahENaC, and that both the N terminus and the second transmembrane segment (M2) were required for gamma subunit function. The biochemical studies were supported by functional studies. Co-expression of gamma subunits lacking M2 with full-length hENaC subunits revealed an inhibitory effect on hENaC channel function that appeared to be mediated by the cytoplasmic N terminus of gamma, and was consistent with the assembly of nonfunctional subunits into the channel complex. We conclude that the N terminus of gammahENaC is involved in channel assembly. PMID- 9341176 TI - Biosynthesis of the phagocyte NADPH oxidase cytochrome b558. Role of heme incorporation and heterodimer formation in maturation and stability of gp91phox and p22phox subunits. AB - The NADPH oxidase cytochrome b558 is a membrane heterodimer comprised of a glycosylated 91-kDa subunit, gp91(phox), and a nonglycosylated 22-kDa subunit, p22(phox). The role of heme in cytochrome b558 biosynthesis was studied using succinyl acetone, an inhibitor of heme synthesis, in PLB-985 myeloid cells undergoing granulocytic differentiation. Succinyl acetone markedly reduced expression of p22(phox) and the mature 91-kDa form of gp91(phox) but not its 65 kDa high mannose precursor, in association with a profound reduction in NADPH oxidase activity. Expression of non-heme-containing cytosolic oxidase components was unaffected. The reduction in cytochrome b558 expression and NADPH oxidase activity was prevented by adding exogenous heme and was reversible upon removal of succinyl acetone. Transgenic expression of gp91(phox) in monkey COS-7 and murine 3T3 cells, both of which lacked endogenous p22(phox) mRNA, demonstrated that p22(phox) was not required for maturation of gp91(phox) carbohydrate to complex oligosaccharides. However, coexpression of transgenic p22(phox) increased the abundance of the mature gp91(phox) glycoprotein. These results suggest that heme incorporation plays an important role in cytochrome b558 assembly and provide further support for the concept that stability of p22(phox) and the mature gp91(phox) subunit is increased by heterodimer formation. PMID- 9341178 TI - Alpha-catenin can form asymmetric homodimeric complexes and/or heterodimeric complexes with beta-catenin. AB - The cadherin-based transmembrane cell-cell adhesive complex is thought to be composed of a cadherin molecule, a beta-catenin, and an alpha-catenin, which connects the complex to the cytoskeleton. The precise stoichiometry of this complex remains uncertain. We have used a series of recombinant molecules and biophysical techniques to assess the multimeric state of human alpha- and beta catenin in vitro and then visualized them by electron microscopy after rotary shadowing. Calculated solution molecular masses are 213 kDa for alpha-catenin, 73 kDa for beta-catenin, and 186 kDa for both. This suggests that alpha-catenin exists as a homodimer in solution, beta-catenin is a monomer, and when both are present, they form alpha/beta-catenin heterodimers. Co-precipitation and surface plasmon resonance assays localize the site of alpha-catenin dimerization to the NH2-terminal 228 amino acids. This region encompasses a high-affinity (Kd = 100 nM) binding site for beta-catenin that lies between residues 54 and 157. We anticipate that the oligomeric state of alpha-catenin and the relative stoichiometry of the components in the membrane adhesion complex will be dynamic and regulated by beta-catenin, cell adhesion, and probably other factors as well. PMID- 9341179 TI - Alterations in lipoprotein metabolism in peroxisome proliferator-activated receptor alpha-deficient mice. AB - The peroxisome proliferator-activated receptor-alpha (PPARalpha) controls gene expression in response to a diverse class of compounds collectively referred to as peroxisome proliferators. Whereas most known peroxisome proliferators are of exogenous origin and include hypolipidemic drugs and other industrial chemicals, several endogenous PPARalpha activators have been identified such as fatty acids and steroids. The latter finding and the fact that PPARalpha modulates target genes encoding enzymes involved in lipid metabolism suggest a role for PPARalpha in lipid metabolism. This was investigated in the PPARalpha-deficient mouse model. Basal levels of total serum cholesterol, high density lipoprotein cholesterol, hepatic apolipoprotein A-I mRNA, and serum apolipoprotein A-I in PPARalpha-deficient mice are significantly higher compared with wild-type controls. Treatment with the fibrate Wy 14,643 decreased apoA-I serum levels and hepatic mRNA levels in wild-type mice, whereas no effect was detected in the PPARalpha-deficient mice. Administration of the fibrate Wy 14,643 to wild-type mice results in marked depression of hepatic apolipoprotein C-III mRNA and serum triglycerides compared with untreated controls. In contrast, PPARalpha-deficient mice were unaffected by Wy 14,643 treatment. These studies demonstrate that PPARalpha modulates basal levels of serum cholesterol, in particular high density lipoprotein cholesterol, and establish that fibrate-induced modulation in hepatic apolipoprotein A-I, C-III mRNA, and serum triglycerides observed in wild-type mice is mediated by PPARalpha. PMID- 9341181 TI - Mutational analysis of conserved residues of the beta-subunit of human farnesyl:protein transferase. AB - The roles of 11 conserved amino acids of the beta-subunit of human farnesyl:protein transferase (FTase) were examined by performing kinetic and biochemical analyses of site-directed mutants. This biochemical information along with the x-ray crystal structure of rat FTase indicates that residues His-248, Arg-291, Lys-294, and Trp-303 are involved with binding and utilization of the substrate farnesyl diphosphate. Our data confirm structural evidence that amino acids Cys-299, Asp-297, and His-362 are ligands for the essential Zn2+ ion and suggest that Asp-359 may also play a role in Zn2+ binding. Additionally, we demonstrate that Arg-202 is important for binding the essential C-terminal carboxylate of the protein substrate. PMID- 9341180 TI - Phosphorylation of chemoattractant receptors is not essential for chemotaxis or termination of G-protein-mediated responses. AB - In several G-protein-coupled signaling systems, ligand-induced receptor phosphorylation by specific kinases is suggested to lead to desensitization via mechanisms including receptor/G-protein uncoupling, receptor internalization, and receptor down-regulation. We report here that elimination of phosphorylation of a chemoattractant receptor of Dictyostelium, either by site-directed substitution of the serines or by truncation of the C-terminal cytoplasmic domain, completely prevented agonist-induced loss of ligand binding but did not impair the adaptation of several receptor-mediated responses including the activation of adenylyl and guanylyl cyclases and actin polymerization. In addition, the phosphorylation-deficient receptors were capable of mediating chemotaxis, aggregation, and differentiation. We propose that for chemoattractant receptors agonist-induced phosphorylation regulates surface binding activity but other phosphorylation-independent mechanisms mediate response adaptation. PMID- 9341182 TI - Overexpression, purification, characterization, and crystallization of the BTB/POZ domain from the PLZF oncoprotein. AB - The BTB/POZ domain defines a conserved region of about 120 residues and has been found in over 40 proteins to date. It is located predominantly at the N terminus of Zn-finger DNA-binding proteins, where it may function as a repression domain, and less frequently in actin-binding and poxvirus-encoded proteins, where it may function as a protein-protein interaction interface. A prototypic human BTB/POZ protein, PLZF (promyelocytic leukemia zinc finger) is fused to RARalpha (retinoic acid receptor alpha) in a subset of acute promyelocytic leukemias (APLs), where it acts as a potent oncogene. The exact role of the BTB/POZ domain in protein protein interactions and/or transcriptional regulation is unknown. We have overexpressed, purified, characterized, and crystallized the BTB/POZ domain from PLZF (PLZF-BTB/POZ). Gel filtration, dynamic light scattering, and equilibrium sedimentation experiments show that PLZF-BTB/POZ forms a homodimer with a Kd below 200 nM. Differential scanning calorimetry and equilibrium denaturation experiments are consistent with the PLZF-BTB/POZ dimer undergoing a two-state unfolding transition with a Tm of 70.4 degrees C, and a DeltaG of 12.8 +/- 0.4 kcal/mol. Circular dichroism shows that the PLZF-BTB/POZ dimer has significant secondary structure including about 45% helix and 20% beta-sheet. We have prepared crystals of the PLZF-BTB/POZ that are suitable for a high resolution structure determination using x-ray crystallography. The crystals form in the space group I222 or I212121 with a = 38.8, b = 77.7, and c = 85.3 A and contain 1 protein subunit per asymmetric unit with approximately 40% solvent. Our data support the hypothesis that the BTB/POZ domain mediates a functionally relevant dimerization function in vivo. The crystal structure of the PLZF-BTB/POZ domain will provide a paradigm for understanding the structural basis underlying BTB/POZ domain function. PMID- 9341183 TI - Activation of endothelin ETA receptors induces phosphorylation of alpha1b adrenoreceptors in Rat-1 fibroblasts. AB - The effect of endothelin-1 on the phosphorylation of alpha1b-adrenoreceptors, transfected into rat-1 fibroblasts, was studied. Basal alpha1b-adrenoreceptor phosphorylation was markedly increased by endothelin-1, norepinephrine, and phorbol esters. The effect of endothelin-1 was dose dependent (EC50 approximately 1 nM), reached its maximum 5 min after stimulation, and was inhibited by BQ-123, an antagonist selective for ETA receptors. Endothelin-1-induced alpha1b adrenoreceptor phosphorylation was attenuated by staurosporine or genistein and essentially abolished when both inhibitors were used together. The effect of norepinephrine was not modified by either staurosporine or genistein alone, and it was only partially inhibited when both were used together. These data suggest the participation of protein kinase C and tyrosine kinase(s) in endothelin-1 induced receptor phosphorylation. However, phosphoaminoacid analysis revealed the presence of phosphoserine and traces of phosphothreonine, but not of phosphotyrosine, suggesting that the putative tyrosine kinase(s), activated by endothelin, could act in a step previous to receptor phosphorylation. The effect of endothelin-1 on alpha1b-adrenoreceptor phosphorylation was not mediated through pertussis toxin-sensitive G proteins. Calcium mobilization induced by norepinephrine was diminished by endothelin-1. Norepinephrine and endothelin-1 increased [35S]GTPgammaS binding to control membranes. The effect of norepinephrine was abolished in membranes obtained from cells pretreated with endothelin-1. Interestingly, genistein plus staurosporine inhibited this effect of the endothelial peptide. Endothelin-1 did not induce alpha1b-adrenoreceptor internalization. Our data indicate that activation of ETA receptors by endothelin 1 induces alpha1b-adrenoreceptor phosphorylation and alters G protein coupling. PMID- 9341184 TI - An oxidative damage-specific endonuclease from rat liver mitochondria. AB - Reactive oxygen species have been shown to generate mutagenic lesions in DNA. One of the most abundant lesions in both nuclear and mitochondrial DNA is 7,8-dihydro 8-oxoguanine (8-oxoG). We report here the partial purification and characterization of a mitochondrial oxidative damage endonuclease (mtODE) from rat liver that recognizes and incises at 8-oxoG and abasic sites in duplex DNA. Rat liver mitochondria were purified by differential and Percoll gradient centrifugation, and mtODE was extracted from Triton X-100-solubilized mitochondria. Incision activity was measured using a radiolabeled double-stranded DNA oligonucleotide containing a unique 8-oxoG, and reaction products were separated by polyacrylamide gel electrophoresis. Gel filtration chromatography predicts mtODE's molecular mass to be between 25 and 30 kDa. mtODE has a monovalent cation optimum between 50 and 100 mM KCl and a pH optimum between 7.5 and 8. mtODE does not require any co-factors and is active in the presence of 5 mM EDTA. It is specific for 8-oxoG and preferentially incises at 8-oxoG:C base pairs. mtODE is a putative 8-oxoG glycosylase/lyase enzyme, because it can be covalently linked to the 8-oxoG oligonucleotide by sodium borohydride reduction. Comparison of mtODE's activity with other known 8-oxoG glycosylases/lyases and mitochondrial enzymes reveals that this may be a novel protein. PMID- 9341185 TI - 20-Hydroxyeicosatetraenoic acid-induced vasoconstriction and inhibition of potassium current in cerebral vascular smooth muscle is dependent on activation of protein kinase C. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE), a cytochrome P450 metabolite of arachidonic acid, is a potent vasoconstrictor, and has been implicated in the myogenic activation of renal and cerebral arteries. We examined the role of protein kinase C (PKC) in the signal transduction pathway by which 20-HETE induces vasoconstriction and inhibition of whole-cell K+ current in cat cerebral vascular smooth muscle. 20-HETE induced a concentration-dependent constriction in isolated pressurized cat middle cerebral arteries (-29 +/- 8% at 1 microM). However, in the presence of an N-myristoylated PKC pseudosubstrate inhibitor peptide (MyrPsiPKC-I(19-27)), 20-HETE induced a concentration-dependent vasodilation (26 +/- 4% at 1 microM). In whole-cell voltage clamp studies, application of 20-HETE inhibited whole-cell K+ current recorded in cat cerebral vascular smooth muscle cells, an effect that was attenuated by MyrPsiPKC-I(19 27). Further evidence for the role of PKC activation in response to 20-HETE is the finding that 20-HETE increased the phosphorylation of myristoylated, alanine rich PKC substrate in cultured cat cerebral vascular smooth muscle cells in a concentration- and PKC-dependent manner. These data provide evidence that PKC is an integral part of the signal transduction pathway by which 20-HETE elicits vasoconstriction of cerebral arteries and inhibition of whole-cell K+ current in cat cerebral vascular smooth muscle. PMID- 9341186 TI - O-Glycosylation of C-terminal tandem-repeated sequences regulates the secretion of rat pancreatic bile salt-dependent lipase. AB - Amino acid sequences rich in Pro, Glu, Ser, and Thr (PEST) are common to rapidly degraded proteins (Rogers, S., Wells, R. & Rechsteiner, M. (1986) Science 234, 364-368). On pancreatic bile salt-dependent lipase (BSDL), PEST sequences are present in the C-terminal region of the enzyme to which is associated the O glycosylation. We have postulated that the O-glycosylation of BSDL may contribute to mask PEST sequences and to trigger the secretion of this enzyme instead of its delivery into a degradative pathway (Bruneau, N., and Lombardo, D. (1995) J. Biol. Chem. 270, 13524-13523). To further examine the role of the O-linked glycosylation on BSDL metabolism, rat pancreatic BSDL cDNA was stably transfected into two Chinese hamster ovary (CHO) cell lines, the CHO K1 wild-type line and the O-glycosylation defective CHO ldlD line. In these latter cells, O glycosylation can be reversibly modulated by culture conditions. Results indicate that the rate of BSDL synthesis by transfected CHO K1 or CHO ldlD cells reflects, independently of culture conditions, the amount of mRNA specific for BSDL present in these transfected cells. Nevertheless, the rate of secretion of the enzyme depends upon cell culture conditions and increases with the cell capability to O glycosylate C-terminal tandem-repeated sequences. Immunoprecipitation experiments performed on cell lysates suggested that a rapid degradation of BSDL occurred particularly when transfected CHO ldlD cells were cultured under non-permissive conditions. We further showed that BSDL secreted by CHO ldlD cells grown under non-permissive conditions that normally prevent O-glycosylation incorporated galactose and was reactive with peanut agglutinin, which recognizes the core structure of O-linked glycans. We concluded that the BSDL expressed by CHO ldlD cells grown under non-permissive conditions was rapidly degraded but a fraction of the enzyme was allowed to O-glycosylate and consequently was secreted. PMID- 9341187 TI - Identification of two tyrosine phosphoproteins, pp70 and pp68, which interact with phospholipase Cgamma, Grb2, and Vav after B cell antigen receptor activation. AB - Tyrosine phosphorylation of cellular proteins mediates the assembly and localization of effector proteins through interactions facilitated by modular Src homology 2 (SH2) and phosphotyrosine binding domains. We describe here two tyrosine-phosphorylated proteins with Mr values of 70,000 and 68,000 that interact with Grb2, phospholipase C (PLCgamma1 and PLCgamma2), and Vav after B cell receptor cross-linking. The interaction of pp70 and pp68 with PLC and Vav is mediated by the carboxyl-terminal SH2 domain of PLC and the SH2 domain of Vav. In contrast, the interaction of pp70 and pp68 with Grb2 requires cooperative binding of the SH2 and SH3 domains of Grb2. Western blot analysis demonstrated that neither pp70 nor pp68 represented the recently described linker protein SLP-76, which binds Grb2, PLC, and Vav in T cells after T cell receptor activation. Moreover, SLP-76 protein was not detected in a number of B cell lines or in normal mouse B cells. Hence, we propose that pp70 and pp68 likely represent B cell homologs of SLP-76 which facilitate and coordinate B cell activation. PMID- 9341188 TI - The prooncoprotein EWS binds calmodulin and is phosphorylated by protein kinase C through an IQ domain. AB - A growing family of proteins is regulated by protein kinase C and calmodulin through IQ domains, a regulatory motif originally identified in neuromodulin (Alexander, K. A., Wakim, B. T., Doyle, G. S., Walsh, K. A., and Storm, D. R. (1988) J. Biol. Chem. 263, 7544-7549). Here we report that EWS, a nuclear RNA binding prooncoprotein, contains an IQ domain, is phosphorylated by protein kinase C, and interacts with calmodulin. Interestingly, PKC phosphorylation of EWS inhibits its binding to RNA homopolymers, and conversely, RNA binding to EWS interferes with PKC phosphorylation. Several other RNA-binding proteins, including TLS/FUS and PSF, co-purify with EWS. PKC phosphorylation of these proteins also inhibits their binding to RNA in vitro. These data suggest that PKC may regulate interactions of EWS and other RNA-binding proteins with their RNA targets and that IQ domains may provide a regulatory link between Ca2+ signal transduction pathways and RNA processing. PMID- 9341189 TI - Cloning and expression of a gene encoding a protein obtained from earthworm secretion that is a chemoattractant for garter snakes. AB - The protein ES20, derived from earthworm shock secretion, is a vomeronasally mediated chemoattractant for garter snakes (Jiang, X. C., Inouchi, J., Wang, D., and Halpern, M. (1990) J. Biol. Chem. 265, 8736-8744). Based on its 15-residue N terminal amino acid sequence, degenerative oligodeoxynucleotide probes were synthesized and used to screen a cDNA library that was constructed in sense orientation using a Uni-ZAPTM XR vector and XL1-Blue MRF' host. A gene was cloned from a polymerase chain reaction as well as from the cDNA library. A combination of the forward degenerative primer and T7 primer was used to obtain gene-specific DNA fragments, from which probes were synthesized and successfully used in screening the cDNA library. The ES20 gene is about 700 base pairs long and encodes 208 amino residues. The ES20 gene was excised from a recombinant plasmid pSK-ES20, ligated to pQE30 expression vector, and transformed into Escherichia coli strain JM109. The selected recombinant plasmids were transformed into expression host cell, E. coli M15[pREP4]. Three transformants were selected, induced with isopropyl-1-thio-beta-D-galactopyranoside for fusion gene expression and an expressed 20-kDa fusion protein purified under denaturing conditions. This protein was refolded and gave a positive reaction against ES20-specific polyclonal antibodies. The fusion protein that had not been denatured remained as an aggregate and was an active chemoattractant for garter snakes. PMID- 9341190 TI - The homeodomain protein Arix interacts synergistically with cyclic AMP to regulate expression of neurotransmitter biosynthetic genes. AB - Transcription of the neurotransmitter biosynthetic genes tyrosine hydroxylase and dopamine beta-hydroxylase (DBH) is regulated by cell type-specific transcription factors, including the homeoprotein Arix, and second messengers, including cyclic AMP. The cis-acting regulatory sites of the DBH gene which respond to Arix and cAMP lie adjacent to each other, between bases -180 and -150, in a regulatory element named DB1. Neither Arix nor cyclic AMP analogs alone effectively stimulate transcription from the DBH promoter in non-neuronal cell cultures. However, when Arix is present together with cAMP, transcription is substantially activated. Synergistic transcription from the DBH promoter can also be elicited by cotransfection of Arix with an expression vector encoding the catalytic subunit of protein kinase A. Nuclear extracts from PC12 cells display a cAMP induced complex binding to the DB1 element, and antisera to transcription factors CREB, CREM, Fos, and Jun indicate that these proteins, or closely related family members, interact with DB1. A dominant negative construct of CREB inhibits the response of the DBH promoter to protein kinase A. These results demonstrate a synergistic interaction between a homeodomain protein and the cAMP signal transduction system and suggest that similar interactions may regulate the tissue specific expression of neuroendocrine genes. PMID- 9341191 TI - Plasma phospholipid transfer protein. Adenovirus-mediated overexpression in mice leads to decreased plasma high density lipoprotein (HDL) and enhanced hepatic uptake of phospholipids and cholesteryl esters from HDL. AB - In vitro studies have shown that plasma phospholipid transfer protein (PLTP) converts isolated human high density lipoprotein-3 (HDL3) into larger HDL particles and generates lipid-poor apoA-I containing nascent HDL. To evaluate the role of PLTP in vivo we generated recombinant adenovirus vectors containing either human PLTP cDNA (rPLTP.AdV) or the reporter luciferase cDNA as a control. After intravenous infusion of 4 x 10(7) plaque-forming units (low dose) and 4 x 10(8) plaque-forming units (high dose) of rPLTP.AdV into mice, PLTP activity in plasma increased from base-line levels of 8.4 +/- 0.2 to 108 +/- 17 and from 8.9 +/- 0.6 to 352 +/- 31 micromol/ml/h, respectively, on day 4 (both p < 0.001). Thus, both low and high doses of rPLTP.AdV led to pronounced overexpression of human PLTP in mice. On day 4 after treatment, mice treated with low and high doses of rPLTP.AdV showed decreased HDL cholesterol (-54% and -91%) and apoA-I ( 64% and -98%) (all p < 0.05). Kinetic studies revealed that the fractional catabolic rates of HDL labeled with [3H]phosphatidylcholine, [14C]phosphatidylcholine ether, [3H]cholesteryl ether, and 125I-labeled mouse apoA-I were increased by 8.5-, 8.7-, 3.8-, and 2.8-fold, respectively, in mice treated with low dose rPLTP.AdV (all p < 0.001). After injection of labeled HDL, mice treated with rPLTP.AdV showed an increased accumulation of labeled PC ether (+304%) and cholesteryl ether (+92%) in the liver (both p < 0.05). Two dimensional gel electrophoresis of plasma 5 min after injection of HDL labeled with 125I-apoA-I demonstrated increased levels of newly generated pre-beta-HDL in mice overexpressing PLTP. In conclusion, HDL remodeling mediated by PLTP generates nascent, lipid-poor apoA-I in vivo and accelerates the hepatic uptake of HDL surface and core lipids in mice treated with rPLTP.AdV. Accelerated catabolism of HDL in mice overexpressing PLTP leads to low HDL levels. Our data indicate an important role for PLTP in modulating reverse cholesterol transport in vivo. PMID- 9341192 TI - Osmotic shock stimulates GLUT4 translocation in 3T3L1 adipocytes by a novel tyrosine kinase pathway. AB - Similar to insulin, osmotic shock of 3T3L1 adipocytes stimulated an increase in glucose transport activity and translocation of GLUT4 protein from intracellularly localized vesicles to the plasma membrane. The docking/fusion of GLUT4 vesicles with the plasma membrane appeared to utilize a similar mechanism, since expression of a dominant interfering mutant of syntaxin-4 prevented both insulin- and osmotic shock-induced GLUT4 translocation. However, although the insulin stimulation of GLUT4 translocation and glucose transport activity was completely inhibited by wortmannin, activation by osmotic shock was wortmannin insensitive. Furthermore, insulin stimulated the phosphorylation and activation of the Akt kinase, whereas osmotic shock was completely without effect. Surprisingly, treatment of cells with the tyrosine kinase inhibitor, genistein, or microinjection of phosphotyrosine antibody prevented both the insulin- and osmotic shock-stimulated translocation of GLUT4. In addition, osmotic shock induced the tyrosine phosphorylation of several discrete proteins including Cbl, p130(cas), and the recently identified soluble tyrosine kinase, calcium-dependent tyrosine kinase (CADTK). In contrast, insulin had no effect on CADTK but stimulated the tyrosine phosphorylation of Cbl and the tyrosine dephosphorylation of pp125(FAK) and p130(cas). These data demonstrate that the osmotic shock stimulation of GLUT4 translocation in adipocytes occurs through a novel tyrosine kinase pathway that is independent of both the phosphatidylinositol 3-kinase and the Akt kinase. PMID- 9341193 TI - The tax protein of human T-cell leukemia virus type 1 mediates the transactivation of the c-sis/platelet-derived growth factor-B promoter through interactions with the zinc finger transcription factors Sp1 and NGFI-A/Egr-1. AB - Transcriptional up-regulation of the c-sis/platelet-derived growth factor-B (PDGF B) proto-oncogene by the Tax protein of human T-cell leukemia virus type 1 has been implicated as one possible mechanism of cellular transformation by human T cell leukemia virus type 1. In previous work, we identified an essential site in the c-sis/PDGF-B promoter, Tax-responsive element 1 (TRE1), necessary for transactivation by Tax. We also identified Sp1, Sp3, and NGFI-A/Egr-1 as the primary nuclear transcription factors binding to TRE1 which mediate Tax responsiveness. In the present work, we have investigated the mechanism(s) whereby Tax transactivates the c-sis/PDGF-B proto-oncogene. In vitro transcription assays showed that Tax was able to significantly increase the transcriptional activity of a template containing the -257 to +74 region of the c sis/PDGF-B promoter. Electrophoretic mobility shift assay analysis showed that Tax increased the DNA binding activity of both Sp1 and NGFI-A/Egr-1 using a TRE1 probe. Analysis of Tax mutants showed that two mutants, IEXC29S and IEXL320G, were unable to significantly transactivate the c-sis/PDGF-B promoter. Finally, co immunoprecipitation analysis revealed that Tax is able to stably bind to both Sp1 and NGFI-A/Egr-1. Interestingly, co-immunoprecipitation analysis also revealed that Tax mutant IEXC29S is unable to interact with NGFI-A/Egr-1, whereas Tax mutant IEXL320G is able to interact with NGFI-A/Egr-1. PMID- 9341194 TI - Altered activity of palmitoylation-deficient and isoprenylated forms of the G protein-coupled receptor kinase GRK6. AB - G protein-coupled receptor kinases (GRKs) utilize diverse mechanisms to associate with the plasma membrane and mediate phosphorylation of agonist-occupied receptors. For example, two members of this family, GRK4 and GRK6, contain C terminal cysteine residues that are palmitoylated. To address whether the activity and membrane association of GRK6 is regulated by palmitoylation, we overexpressed and characterized wild-type GRK6 and two GRK6 mutants, one with the palmitoylation sites mutated to serines (GRK6-pal-) and one containing a C terminal CAAX motif to promote geranylgeranylation (GRK6-GG). Compared with wild type GRK6, GRK6-pal- had a approximately 5-fold higher Km and approximately 2 fold lower Vmax for phosphorylating rhodopsin, whereas GRK6-GG exhibited a approximately 2-fold lower Km and approximately 14-fold higher Vmax for rhodopsin. In contrast, wild-type GRK6 and GRK6-pal- displayed similar activity toward the nonreceptor substrate phosvitin, indicating that nonpalmitoylated GRK6 is catalytically active. Wild-type GRK6 and GRK6-GG, but not GRK6-pal-, also bound significantly to phosphatidylcholine vesicles (36 +/- 3, 79 +/- 4, and 4 +/ 2%, respectively) suggesting that GRK6 activity is dependent upon its ability to interact with the plasma membrane. When assayed in COS-1 cells GRK6-pal- promoted minimal agonist-dependent sequestration of the beta2-adrenergic receptor, while sequestration was significantly increased in cells expressing either wild-type GRK6 or GRK6-GG. These data demonstrate an important functional link between the ability of GRK6 to bind to the plasma membrane, a process that appears to be regulated by palmitoylation, and its activity toward receptor substrates. PMID- 9341195 TI - Identification and functional requirement of Cu(I) and its ligands within coagulation factor VIII. AB - Coagulation factor VIII (FVIII) is a heterodimer consisting of a light chain of 80 kDa (domains A3-C1-C2) in a metal ion-dependent association with a 220-kDa heavy chain (domains A1-A2-B). The nature of the metal ion-dependent association between the heavy and light chains was investigated using atomic absorption spectroscopy, electron paramagnetic resonance spectroscopy (EPR), and site directed mutagenesis and expression of the FVIII cDNA. Whereas copper ion was not detected in intact recombinant FVIII, EDTA dissociation of the chains yielded an EPR signal consistent with 1 mol of Cu(I)/mol of active protein, supporting the hypothesis that a single molecule of reduced copper ion is buried within intact FVIII and is released and oxidized upon treatment with EDTA. Cu(I), and not Cu(II), was able to reconstitute FVIII activity from dissociated chains, demonstrating a requirement for Cu(I) in FVIII function. Three potential copper ion binding sites exist within FVIII: one type-2 site and two type-1 sites. The importance of these potential copper ion ligands was tested by studying the effect of site-directed mutants. Of the two histidines that compose the type-2 binding site, the His-1957 --> Ala mutant displayed secretion, light and heavy chain assembly, and activity similar to wild-type FVIII, while mutant His-99 --> Ala was partially defective for secretion and had low levels of heavy and light chain association and activity. In contrast, FVIII having the mutation Cys-310 - > Ser within the type-1 copper binding site in the A1 domain was inactive and partially defective for secretion from the cell, and the heavy and light chains of the secreted protein were not associated. Mutant Cys-2000 --> Ser within the A3 domain displayed secretion, assembly, and activity similar to that for wild type FVIII. These results support the hypothesis that Cu(I) is buried within the type-1 copper binding site within the A1 domain and is required for FVIII chain association and activity. PMID- 9341196 TI - The interaction between the AsiA protein of bacteriophage T4 and the sigma70 subunit of Escherichia coli RNA polymerase. AB - The AsiA protein of bacteriophage T4 binds to the sigma70 subunit of Escherichia coli RNA polymerase and plays a dual regulatory role during T4 development: (i) inhibition of host and phage early transcription, and (ii) coactivation of phage middle-mode transcription, which also requires the T4 DNA binding transcriptional activator, MotA. We report that the interaction between AsiA and sigma70 occurs with a 1:1 stoichiometry. When preincubated with RNA polymerase, AsiA is a potent inhibitor of open complex formation at the lac UV5 promoter, whereas it does not perturb preformed open or intermediate promoter complexes. DNase I footprinting and electrophoretic mobility shift analyses of RNA polymerase-DNA complexes formed at the T4 early promoter P15.0 show that AsiA blocks the initial RNA polymerase binding step that leads to the formation of specific closed promoter complexes. A contrasting result is obtained on the T4 middle promoter PrIIB2, where AsiA stimulates the formation of both closed complexes and open complexes. Therefore, we propose that AsiA modulates initial DNA binding by the RNA polymerase, switching promoter usage at the level of closed complex formation. PMID- 9341197 TI - Identification of elongin C sequences required for interaction with the von Hippel-Lindau tumor suppressor protein. AB - Elongin C is a 112-amino acid protein that is found in mammalian cells as a positive regulatory subunit of heterotrimeric RNA polymerase II elongation factor Elongin (SIII) and as a component of a multiprotein complex containing the von Hippel-Lindau (VHL) tumor suppressor protein. As a subunit of the Elongin complex, Elongin C interacts directly with the transcriptionally active Elongin A subunit and potently induces its elongation activity; in addition, Elongin C interacts with the ubiquitin-like Elongin B subunit, which regulates the interaction of Elongin C with Elongin A. As a component of the VHL complex, Elongin C interacts directly with both Elongin B and the VHL protein. Binding of the VHL protein to Elongin C was found to prevent Elongin C from interacting with and activating Elongin A in vitro, leading to the proposal that one function of the VHL protein may be to regulate RNA polymerase II elongation by negatively regulating the Elongin complex. In this report, we identify Elongin C sequences required for its interaction with the VHL protein. We previously demonstrated that the ability of Elongin C to bind and activate Elongin A is sensitive to mutations in the C-terminal half of Elongin C, as well as to mutations in an N terminal Elongin C region needed for formation of the Elongin BC complex. Here we show that interaction of Elongin C with the VHL tumor suppressor protein depends strongly on sequences in the C terminus of Elongin C but is independent of the N terminal Elongin C region required for binding to Elongin B and for binding and activation of Elongin A. Taken together, our results are consistent with the proposal that the VHL protein negatively regulates Elongin C activation of the Elongin complex by sterically blocking the interaction of C-terminal Elongin C sequences with Elongin A. In addition, our finding that only a subset of Elongin C sequences required for its interaction with Elongin A are critical for binding to VHL may offer the opportunity to develop reagents that selectively interfere with Elongin and VHL function. PMID- 9341198 TI - Lyn associates with the juxtamembrane region of c-Kit and is activated by stem cell factor in hematopoietic cell lines and normal progenitor cells. AB - Stem cell factor (SCF) is a cytokine critical for normal hematopoiesis. The receptor for SCF is c-Kit, a receptor tyrosine kinase. Our laboratory is interested in delineating critical components of the SCF signal transduction pathway in hematopoietic tissue. The present study examines activation of Src family members in response to SCF. Stimulation of cell lines as well as normal progenitor cells with SCF rapidly increased tyrosine phosphorylation of the Src family member Lyn. Peak responses were noted 10-20 min after SCF treatment, and phosphorylation of Lyn returned to basal levels 60-90 min after stimulation. SCF also induced increases in Lyn kinase activity in vitro. Lyn coimmunoprecipitated with c-Kit, and studies with GST fusion proteins demonstrated that Lyn readily associated with the juxtamembrane region of c-Kit. Treatment of cells with either Lyn antisense oligonucleotides or PP1, a Src family inhibitor, resulted in dramatic inhibition of SCF-induced proliferation. These data demonstrate that SCF rapidly activates Lyn and suggest that Lyn is critical in SCF-induced proliferation in hematopoietic cells. PMID- 9341199 TI - Molecular cloning and expression of palmitoyl-protein thioesterase 2 (PPT2), a homolog of lysosomal palmitoyl-protein thioesterase with a distinct substrate specificity. AB - Palmitoyl-protein thioesterase is a lysosomal hydrolase that removes long chain fatty acyl groups from modified cysteine residues in proteins. Mutations in this enzyme were recently shown to underlie the hereditary neurodegenerative disorder, infantile neuronal ceroid lipofuscinosis, and lipid thioesters derived from acylated proteins were found to accumulate in lymphoblasts from individuals with the disorder. In the current study, we describe the cloning and expression of a second lysosomal thioesterase, palmitoyl-protein thioesterase 2 (PPT2), that shares an 18% identity with palmitoyl-protein thioesterase. Transient expression of a PPT2 cDNA led to the production of a glycosylated lysosomal protein with palmitoyl-CoA hydrolase activity comparable with palmitoyl-protein thioesterase. However, PPT2 did not remove palmitate groups from palmitoylated proteins that are substrates for palmitoyl-protein thioesterase. In cross-correction experiments, PPT2 did not abolish the accumulation of protein-derived lipid thioesters in palmitoyl-protein thioesterase-deficient cell lines. These results indicate that PPT2 is a lysosomal thioesterase that possesses a substrate specificity that is distinct from that of palmitoyl-protein thioesterase. PMID- 9341200 TI - Calcium-stimulated phosphorylation of MAP-2 in pancreatic betaTC3-cells is mediated by Ca2+/calmodulin-dependent kinase II. AB - An understanding of the role of CaM kinase II in the pancreatic beta-cell is dependent on the identification of its cellular targets. One of the best substrates of CaM kinase II in vitro that could function in secretory events is the microtubule-associated protein, MAP-2. By immunoblot analysis, a high molecular weight protein with electrophoretic properties characteristic of MAP-2, was identified in rat insulinoma betaTC3 cells and isolated rat islets. In immunoprecipitation experiments employing alpha-toxin-permeabilized betaTC3 cells, elevation of intracellular Ca2+ or addition of forskolin, an adenylate cyclase activator, induced significant phosphorylation of MAP-2 in situ. The effect of Ca2+ was rapid, concentration-dependent and closely correlated with activation of CaM kinase II under similar experimental conditions. H-89, a specific and potent inhibitor of cAMP-dependent protein kinase (PKA), prevented forskolin-induced MAP-2 phosphorylation but had little effect on MAP-2 phosphorylation stimulated by elevated Ca2+. Phosphopeptide mapping revealed that the phosphorylation pattern observed in situ upon incubation of the betaTC3 cells with increased free Ca2+, was strikingly similar to that generated in vitro by CaM kinase II, most notably with regard to the increased phosphate incorporated into one prominent site. These data provide evidence that MAP-2 is phosphorylated by CaM kinase II in the pancreatic beta-cell in situ, and that this event may provide an important link in the mediation of Ca2+-dependent insulin secretion. PMID- 9341201 TI - A plant small heat shock protein gene expressed during zygotic embryogenesis but noninducible by heat stress. AB - A small heat shock protein (sHSP) gene from sunflower, Ha hsp17.6 G1, showed expression patterns that differ from what is known for members of this gene family. The mRNAs of this gene accumulated in seeds during late desiccation stages of zygotic embryogenesis but not in response to heat shock in vegetative tissues. The failure to respond to heat shock was independent of the developmental stage after germination and shock temperature. Nuclear run-on analyses demonstrated that transcription from the Ha hsp17.6 G1 promoter is not induced by heat shock. This agrees with the presence, in this promoter, of sequences with little similarity to heat shock elements. Our results show an evolutionary divergence, in the regulation of plant sHSP genes, which has originated stress-responsive genes and nonresponsive members within this gene family. We discuss implications for mechanisms controlling the developmental regulation of sHSP genes in plants. PMID- 9341202 TI - Cdc2 and mitogen-activated protein kinases modulate DNA binding properties of the putative transcriptional regulator Chironomus high mobility group protein I. AB - Cells of the dipteran insect Chironomus contain a high mobility group protein that is homologous to the mammalian high mobility group proteins I/Y (HMGI/Y). These proteins facilitate the assembly of higher order nucleoprotein complexes. In proliferating cells, >30% of Chironomus HMGI was found to be phosphorylated. The phosphorylation sites were mapped to Ser3, Ser22, and Ser72 and were found to be substrates for the kinases Cdc2 (and mitogen-activated protein (MAP)), MAP, and Ca2+/phospholipid-dependent protein kinase, respectively. In mitotically arrested cells, the extent of phosphorylation at Ser3 increased, whereas phosphorylation at Ser22 remained unchanged. In nondividing cells, phosphorylation at Ser3 and Ser22 was strongly reduced. The DNA binding affinity of Chironomus HMGI was not influenced by single phosphorylation at Ser3 or Ser22. In contrast, phosphorylation at both of these sites resulted in a 10-fold weakening of the binding activity and altered the mode of protein-DNA interaction. Since both human and murine HMGI/Y proteins, similarly to the insect HMGI protein, possess phosphorylation sites for Cdc2 and MAP kinases that intersperse the AT-hook DNA-binding motifs, our results may reflect a general mechanism that regulates the properties and function of this class of putative transcriptional regulators. PMID- 9341203 TI - Comparison of three members of the cysteine-rich protein family reveals functional conservation and divergent patterns of gene expression. AB - Members of the cysteine-rich protein (CRP) family are evolutionarily conserved proteins that have been implicated in the processes of cell proliferation and differentiation. In particular, one CRP family member has been shown to be an essential regulator of cardiac and skeletal muscle development. Each of the three vertebrate CRP isoforms characterized to date is composed of two copies of the zinc-binding LIM domain with associated glycine-rich repeats. In this study, we have addressed the biological significance of the CRP multigene family by comparing the subcellular distributions, biochemical properties, and expression patterns of CRP1, CRP2, and CRP3/MLP. Our data reveal that all three CRP family members, when expressed in adherent fibroblasts, associate specifically with the actin cytoskeleton. Moreover, all three CRP isoforms are capable of interacting with the cytoskeletal proteins alpha-actinin and zyxin. Together, these observations suggest that CRP family members may exhibit overlapping cellular functions. Differences between the three CRPs are evident in their protein expression patterns in chick embryos. CRP1 expression is detected in a variety of organs enriched in smooth muscle. CRP2 is restricted to arteries and fibroblasts. CRP3/MLP is dominant in organs enriched in striated muscle. CRP isoform expression is also developmentally regulated in the chick. Our findings suggest that the three CRP family members perform similar functions in different muscle derivatives. The demonstration that all members of the CRP family are associated with cytoskeletal components that have been implicated in the assembly and organization of filamentous actin suggests that CRPs contribute to muscle cell differentiation via effects on cytoarchitecture. PMID- 9341204 TI - Site-directed mutants of pseudoazurin: explanation of increased redox potentials from X-ray structures and from calculation of redox potential differences. AB - In order to understand the origins of differences in redox potentials among cupredoxins (small blue type I copper-containing proteins that reversibly change oxidation state and interact with redox partners), we have determined the structures of the native and two mutants (P80A and P80I) of pseudoazurin from Alcaligenes faecalis S-6 in oxidized and reduced forms at resolutions of 2.2 A in the worst case and 1.6 A in the best case. The P80A mutation creates a surface pocket filled by a new water molecule, whereas the P80I mutant excludes this water. Distinct patterns of change occur in response to reduction for all three molecules: the copper position shifts, Met 7 and Pro 35 move, and the relative orientations of residues 81 to 16, 18 to the amide planes of 77 and 86, all change. Systematic changes in the weak electrostatic interactions seen in the structures of different oxidation states can explain the Met 7/Pro 35 structural differences as well as some fluctuating solvent positions. Overall displacement parameters increase reversibly upon reduction. The reduced forms are slightly expanded over the oxidized forms. The geometries of the mutants become more trigonal in their reduced forms, consistent with higher redox potentials (+409 mV for P80A and +450 mV for P80I). Calculations of the differences in redox potentials, using POLARIS, reveal that a water unique to the P80A mutant is required (with correctly oriented hydrogens) to approximate the observed difference in redox potential. The POLARIS calculations suggest that the reduced forms are additionally stabilized through changes in the solvation of the copper center, specifically via the amides of residues 16, 39, 41, 79, and 80 which interact with either Phe 18, Met 86, or Cys 78. The redox potential of P80A is increased largely due to solvation effects, whereas the redox potential of P80I is increased largely due to geometrical effects. PMID- 9341206 TI - Interaction between glycosaminoglycans and immunoglobulin light chains. AB - Amyloidosis is a pathological process in which normally soluble proteins polymerize to form insoluble fibrils (amyloid). Amyloid formation is found in a number of diseases, including Alzheimer's disease, adult-onset diabetes, and light-chain-associated amyloidosis. No pharmaceutical methods currently exist to prevent this process or to remove the fibrils from tissue. The search for treatment and prevention methods is hampered by a limited understanding of the biophysical basis of amyloid formation. Glycosaminoglycans (GAGs) are long, unbranched heteropolysaccharides composed of repeating disaccharide subunits and are known to associate with amyloid fibrils. The interaction of amyloid associated free light chains with GAGs was tested by both size-exclusion high performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis experiments. The results indicated that heparin 16 000 and chondroitin sulfate B and C precipitated both human intact light chains and recombinant light chain variable domains. Although all light chains interacted with heparin, the strongest interactions were obtained with proteins that had formed amyloid. Molecular modeling indicated the possibility of interaction between heparin and the conserved saddlelike surface of the light chain dimer opposite the complementarity-determining segments that form part of the antigen binding site of a functional antibody. This suggestion might offer a new path to block the aggregation of amyloid-associated light chain proteins, by design of antagonists based on properties of GAG binding. A hexasaccharide was modeled as the basis for a possible antagonist. PMID- 9341205 TI - New inhibitors of thrombin and other trypsin-like proteases: hydrogen bonding of an aromatic cyano group with a backbone amide of the P1 binding site replaces binding of a basic side chain. AB - Highly effective thrombin inhibitors have been obtained by preparing boronic acid analogues of m-cyano-substituted phenylalanine and its incorporation into peptides. The cyano group enhances binding by several orders of magnitude. For example, Ac-(D)Phe-Pro-boroPheOH binds to thrombin with a Ki of 320 nM and the Ki of Ac-(D)Phe-Pro-boroPhe(m-CN)-OH is 0.79 nM. Protein crystal structure determination of trypsin complexed to H-(D)Phe-Pro-boroPhe(m-CN)-OH indicates that the aromatic side chain is bound in the P1 binding site and that the cyano group can act as a H-bond acceptor for the amide proton of Gly219. Enhanced binding for inhibitors containing the m-cyano group was observed for coagulation factor Xa and for the factor VIIa.tissue factor complex [Ki values of Ac-(D)Phe Pro-boroPhe(mCN)-OH are 760 and 3.3 nM, respectively]. This result is consistent with the sequence homology of these two enzymes in the P1 binding site. Two enzymes lacking the strict homology in the P1 binding site, pancreatic kallikrein and chymotrypsin, did not exhibit significantly enhanced binding. PMID- 9341207 TI - Fourier transform infrared evidence for connectivity between CuB and glutamic acid 286 in cytochrome bo3 from Escherichia coli. AB - Photodissociation of fully reduced, carbonmonoxy cytochrome bo3 causes ultrafast transfer of carbon monoxide (C triple bond O) from heme iron to CuB in the binuclear site. At low temperatures, the C triple bond O remains bound to CuB for extended times. Here, we show that the binding of C triple bond O to CuB perturbs the IR stretch of an un-ionized carboxylic acid residue, which is identified as Glu286 by mutation to Asp or to Cys. Before photodissociation, the carbonyl (C=O) stretching frequency of this carboxylic acid residue is 1726 cm-1 for Glu286 and 1759 cm-1 for Glu286Asp. These frequencies are definitive evidence for un-ionized R-COOH and suggest that the carboxylic acids are hydrogen-bonded, though more extensively in Glu286. In Glu286Cys, this IR feature is lost altogether. We ascribe the frequency shifts in the C=O IR absorptions to the effects of binding photodissociated C triple bond O to CuB, which are relay ed to the 286 locus. Conversely, the 2065 cm-1 C triple bond O stretch of CuB-CO is markedly affected by both mutations. These effects are ascribed to changes in the Lewis acidity of CuB, or to displacement of a CuB histidine ligand by C triple bond O. C triple bond O binding to CuB also induces a downshift of an IR band which can be attributed to an aromatic C-H stretch, possibly of histidine imidazole, at about 3140 cm-1. The results suggest an easily polarizable, through-bond connectivity between one of the histidine CuB ligands and the carboxylic group of Glu286. A chain of bound water molecules may provide such a connection, which is of interest in the context of the proton pump mechanism of the heme-copper oxidases. PMID- 9341208 TI - Effect of nucleotides and actin on the orientation of the light chain-binding domain in myosin subfragment 1. AB - The X-ray structure of myosin head (S1) reveals the presence of a long alpha helical structure that supports both the essential and the regulatory light chains. It has been proposed that small structural changes in the catalytic domain of S1 are amplified by swinging the long alpha-helix (the "lever arm") to produce approximately 11 nm steps. To probe the spatial position of the putative lever in various S1 states, we have measured, by fluorescence resonance energy transfer (FRET), the effect of nucleotides and actin on the distances between Cys 177 of the essential light chain A1 (which is attached to the alpha-helix) and three loci in the catalytic domain. Cys-177 (donor) was labeled with 1,5-IAEDANS. The trinitrophenylated ADP analog (TNP-ADP, acceptor) was used to measure the distance to the active site. Lys-553 at the actin-binding site, labeled with a fluorescein derivative, and Lys-83 modified with trinitrobenzenesulfonic acid served as two other acceptors. FRET measurements were performed for S1 alone, for its complexes with MgADP and MgATP, for the analogs of the transition state of the ATPase reaction, S1.ADP.BeFx, S1.ADP.AlF4, and S1.ADP.VO4, and for acto-S1 in the absence and in the presence of ADP. When the transition state and acto-S1 complexes were formed, the change in the Cys-177 --> Lys-83 distance was <1.1 A, for the distance Cys-177 --> Lys-553, the change was +/-2.5 A. These distance changes correspond to rotations by <10 degrees and approximately 25 degrees, respectively. For the Cys-177 --> TNP-ADP the interprobe separation decreased by approximately 6 A in the presence of BeFx and AlF4- but only 1.9 A in the presence of vanadate; we do not interpret the 6 A change as resulting from the lever rotation. Using the coordinates of the acto-S1 complex, we have computed the expected changes in these distances resulting from rotation of the lever. These changes were much greater than the ones observed. The above results are inconsistent with models of force generation by S1 in which the head assumes two distinct conformations characterized by large differences in the angle between the motor and the light chain-binding domain. Several alternative mechanisms are proposed. PMID- 9341209 TI - Refolding of urea-denatured tubulin: recovery of nativelike structure and colchicine binding activity from partly unfolded states. AB - Tubulin unfolding in urea proceeds via the formation of a partially unfolded intermediate state, stable in 2 M urea, that unfolds further in higher urea concentrations. The intermediate state had spectroscopic properties reminiscent of a molten globule and negligible colchicine binding activity. Refolding of totally unfolded tubulin in 8 M urea yielded an intermediatelike state characterized by partial burial of tryptophans and partial recovery of secondary and tertiary structures, although colchicine-binding activity of the protein was not regained. Further folding of this intermediatelike state, toward the native conformation, with respect to both structural and functional parameters did not occur. However, a significant percentage of colchicine binding activity and nativelike tertiary structure was recovered when refolding was initiated from partially denatured protein samples, viz., from <1.2 M urea. Thus, although high concentration of urea induced loss of structure and activity was irreversible, the conformational changes induced in restricted regions of tubulin by lower concentrations of urea, which are probably crucial for its various functional properties, could be reversed. PMID- 9341210 TI - Structural polymorphism and dynamism in the DNA segment GATCTTCCCCCCGGAA: NMR investigations of hairpin, dumbbell, nicked duplex, parallel strands, and i motif. AB - Structure and dynamism in a DNA segment d-GATCTTCCCCCCGGAA have been investigated by nuclear magnetic resonance (NMR) spectroscopy. At neutral pH, the molecule exists largely as a dumbbell, formed by the association of two hairpins with sticky ends. A small percentage of hairpin is also detectable. The stem of the dumbbell, which is 12 base pairs long, has two nicks separated by 4 base pairs. The three-dimensional structures of the dumbbell and also of a 12-mer duplex, the sequence of which is identical to that of the stem of the dumbbell, have been determined by NMR in conjunction with restrained molecular dynamics calculations. It is observed that the presence of nicks causes minor changes in the structure of the duplex. Fraying at the nicks is much less than at the ends of a regular duplex. The loop shows very few nuclear Overhauser effects, which is a reflection on the greater dynamism in its structure. At lower pH, the molecule undergoes a transition to an i-motif type of structure with two parallel stranded duplexes involving C-C+ pairing, interdigitating each other. The structure is highly stable, with a melting temperature >60 degrees C. PMID- 9341212 TI - Cleavage of the X-Pro peptide bond by pepsin is specific for the trans isomer. AB - Fluorine nuclear magnetic resonance studies of the cleavage of peptides containing a 4-fluorophenylalanine (FPhe)-Pro bond have been performed in order to determine the conformational specificity of FPhe-Pro bond cleavage by pepsin. The peptides selected were substrates of HIV protease or of avian sarcoma virus protease, both of which have been reported to be cleaved specifically at X-Pro by pepsin as well as by the corresponding viral protease enzyme. By working at 0 degrees C, it was possible to separate kinetically cleavage and cis/trans isomerization. For the case of the protease substrate, Ser-Gln-Asn-FPhe-Pro-Ile Val-Gln, cleavage was shown to be specific for the trans conformation. A value for the rate constant for hydrolysis of the trans peptide divided by the Michaelis constant, ktH/KMtrans = 0.3 min-1 mM-1 was obtained with this substrate, and the Michaelis constant appears to be considerably higher than the substrate concentration, 3.7 mM, used in the study. On a slower time scale, additional cleavages can readily be detected. For the avian leukemia virus protease substrate, Thr-Phe-Gln-Ala-FPhe-Pro-Leu-Arg-Glu-Ala, the cleavage was both slower and less specific. In addition to the primary cleavage at the FPhe Pro site, cleavage also occurs at the Ala-FPhe bond on a somewhat slower time scale. In addition to the conformational specificity of the cleavage reaction, these results indicate that pepsin is a better model for HIV protease than for avian leukemia virus protease. PMID- 9341211 TI - Synergistic inhibition of HIV-1 reverse transcriptase by combinations of chain terminating nucleotides. AB - Synergistic inhibition of HIV replication in cell culture has been reported for many combinations of reverse transcriptase inhibitors. However, the biochemical basis underlying this interaction is in most cases unknown. It has been previously shown that combinations of L-697,661 or U-90152s with AZT or ddC synergistically inhibit HIV-1 replication in cell culture. The combination of AZT with ddC is also favorable with respect to the inhibition of viral replication. However, the corresponding combinations showed no synergy in inhibiting enzyme activity when tested on conventional polymerase assays using homo- or heteropolymeric RNA and DNA as template. Data obtained suggest that amplification of the effect of chain terminators, a consequence of the high potential number of termination sites present on the template, override the synergistic effect expected for the combination of two independent nucleotide analogs. When a saturating amount of enzyme over template:primer was used, and a single site on the template was available for each chain terminator, the combination of AZTTP and ddCTP synergistically inhibited enzyme activity, whereas, as expected, the combination of AZTTP and ddTTP behaved as merely additive. Under similar conditions the combination of U-90152s and AZTTP was also synergistic. These results suggest that synergy found in antiviral assays with combinations having nucleosidic inhibitors is not related to the synergistic inhibition of reverse transcriptase and might be due to the presence in the viral population of virus strains with different sensitivity to the inhibitors. PMID- 9341213 TI - Evidence for Hoogsteen GC base pairs in the proton-induced transition from right handed to left-handed poly(dG-dC).poly(dG-dC). AB - The structure of double-helical poly(dG-dC).poly(dG-dC) is investigated at various pH values with Raman spectroscopy, absorption spectroscopy, and circular dichroism. A comparison is made between the B-form with Watson-Crick base pairing at 1 mM [Na+] and pH 7.2, the Z-form with Watson-Crick base pairing at 4 M [Na+] and pH 7.2, and a different structure at 1 mM [Na+] and pH 4.5 as well as at 150 mM [Na+] and pH 3.1. The CD spectrum of poly(dG-dC). poly(dG-dC) under the latter conditions does not show a negative band at 290 nm. The structure is a double helical structure different from the B-form and the Z-form according to circular dichroism, Raman, and absorption spectroscopic studies. The Raman spectra evidence that the structure contains Hoogsteen base pairing. This can be accommodated in the double helix when the cytosine group is protonated and the sugar-guanine conformer has adopted a C2'-endo/syn conformation. It is shown that this antiparallel-stranded Hoogsteen base paired structure can be maintained under varying conditions, balancing the decrease in pH with an increased salt concentration. It is further concluded that the proton-induced transition from a Watson-Crick to a Hoogsteen base pair is aided by a decrease of [Na+] at pH 4.5 and occurs prior to a conversion from a right-handed helix to a left-handed helix. PMID- 9341214 TI - Evidence that the DNA binding specificity of winged helix proteins is mediated by a structural change in the amino acid sequence adjacent to the principal DNA binding helix. AB - We present the first structural evidence supporting the hypothesis that the binding specificity of the winged helix DNA binding motif is mediated by residues adjacent to the alpha-helix (H3), the moiety which is primarily involved in the interaction with DNA. Using NMR to determine secondary structural elements of a winged helix family member, Genesis (formerly HFH-2), and comparing these with those found in the X-ray crystal structure of the HNF-3gamma/DNA complex [Clark, K. L., Halay, E. D., Lai, E., & Burley, S. K. (1993) Nature 364, 412-420], we show that the major differences observed occur for H3 and the region immediately prior to this. H3 in Genesis is slightly shorter than in HNF-3gamma and, in addition, we observe an extra small helix (H4) in the region between H2 and H3 which is not found in the HNF-3gamma/DNA complex. This is significant as it has been shown previously [Overdier, D. G., Porcella, A., & Costa R. H. (1994) Mol. Cell. Biol. 14, 2755-2766] that the DNA-binding specificity is influenced by amino acid residues in this region. PMID- 9341215 TI - Site-specific phosphorylation of the human immunodeficiency virus type-1 Rev protein accelerates formation of an efficient RNA-binding conformation. AB - Phosphorylation is important in the regulation of many cellular processes, yet the precise role of protein phosphorylation for many RNA-binding protein substrates remains obscure. In this report, we demonstrate that phosphorylation of a recombinant human immunodeficiency virus type-1 Rev protein promotes rapid formation of an efficient RNA-binding state. The apparent dissociation constant for ligand binding is enhanced 7-fold for the protein following phosphorylation; however, phosphate addition leads to a 1. 6-fold decrease in RNA ligand-protein complex stability. RNA ligand binding stimulates slow formation of an equally competent binding state for the unphosphorylated protein, indicating that the addition of phosphate or ligand binding promotes a similar conformational change in Rev. Phosphorylation directly alters the conformation of Rev, as revealed by modification experiments that monitor the solvent accessibility of cysteines in the protein. These biochemical properties are attributed to the addition of phosphate at one of two serine residues (Ser-54 or Ser-56) that lie within the multimerization domain adjacent to the RNA-binding helix. Glutaraldehyde-mediated cross-linking experiments revealed that phosphorylation of Rev does not affect Rev multimerization activity. The Rev protein from the less pathogenic HIV-2 isolate lacks this phosphorylation site in the amino acid sequence; thus, the described biochemical properties of the phosphorylated protein may contribute to Rev activity and possibly to HIV-1 virulence during natural infection. PMID- 9341216 TI - How stereochemistry affects mutagenesis by N2-deoxyguanosine adducts of 7,8 dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene: configuration of the adduct bond is more important than those of the hydroxyl groups. AB - Previous work has shown that the major adduct from the (+)-anti diol epoxide of benzo[a]pyrene (B[a]P), which forms at N2-deoxyguanosine [(+)-trans-anti-B[a]P-N2 dG], is capable of inducing either predominantely G --> T mutations ( approximately 95%) in a 5'-TGC-3 sequence context or predominantly G --> A mutations ( approximately 80%) in a 5'-CGT-3' sequence context. This is likely to be attributable to the major adduct being in a different mutagenic conformation in each case. In the next phase of this work, the questions to be addressed are what conformation is associated with what mutation and why? To help define what aspect of adduct structure is important to mutagenesis, the work herein reports on the mutations induced in a single sequence context by four stereoisomers of B[a]P-N2-dG: (+)-trans-, (+)-cis-, (-)-trans-, and (-)-cis-. The (+)-trans- and ( )-cis-adducts show a remarkably similar mutational pattern with G --> A mutations predominating ( approximately 80%). The (-)-trans- and (+)-cis-adducts also show a similar mutational pattern with a more even mixture of G --> T, G --> A, and G -> C mutations. Each of these adducts has an adduct bond and three hydroxyl groups at four consecutive saturated carbons in the B[a]P moiety of the adduct; the stereochemistry at these four positions differs in each of the adducts. The (+)-trans- and (-)-cis-adducts are a pair sharing the S configuration for the adduct bond, although they are a mirror image vis-a-vis the hydroxyl groups. The (-)-trans- and (+)-cis-adducts share the opposite adduct bond stereochemistry (R) but differ in the stereochemistry of their hydroxyl groups. Thus, there is a correlation suggesting that anti-B[a]P-N2-dG adduct mutagenesis is more dependent on the stereochemistry of the adduct bond than on the stereochemistry of the hydroxyl groups. PMID- 9341217 TI - Purification and characterization of DNA polymerase alpha-associated replication protein A-dependent yeast DNA helicase A. AB - A novel, eukaryotic, hexameric DNA helicase that was earlier identified as a component of the multiprotein polymerase alpha complex [Biswas et al. (1993) Biochemistry 32, 13393-13398] has been purified to homogeneity and characterized. Thus far, our studies demonstrated that helicase A shares certain unique features of two other hexameric DNA helicases: the DnaB helicase of Escherichia coli and the T-antigen helicase of the SV40 virus. The helicase activity was stimulated by yeast replication protein A (RPA) and to a lower extent by E. coli single stranded DNA binding protein (SSB). The helicase had an apparent molecular mass of 90 kDa, as determined by its mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A tryptic peptide fragment of the polypeptide was sequenced followed by a BLAST search of GenBank with the tryptic peptide sequence. The search identified a 1.8 kb open reading frame previously designated as ykl017c on chromosome XI, that codes for a 78.3 kDa (683 amino acid) polypeptide. The important features of the polypeptide sequence of helicase A included a type I ATP/GTP binding motif, and a K E E R R L N V A M T R P R R sequence at the C terminus that may be indicative of a nuclear localization signal which is required of a nuclear DNA helicase. The polypeptide sequence of helicase A appears to have homology to the DnaB helicase of E. coli (approximately 25%). The facts that these two helicases are vastly separated by evolution and retained similar structural and functional features, as demonstrated here, point to a possible significance of this limited homology. Although the amount of purified helicase A was limited, we have carried out necessary enzymatic characterization so that these data could be correlated with that of immunoaffinity-purified helicase A and recombinant helicase A expressed in heterologous systems. PMID- 9341218 TI - Yeast DNA helicase A: cloning, expression, purification, and enzymatic characterization. AB - We have cloned and expressed the yeast DNA helicase A in Escherichia coli at a high level (approximately 30 mg/L of culture) in soluble form. We describe here a simple two-step purification protocol that produces reasonable quantities of homogeneous enzyme. In denaturing gel electrophoresis the enzyme behaved as a approximately 90 kDa protein. The native structure, determined by gel-filtration studies, appeared to be hexameric and its quaternary structure was salt (NaCl) dependent. In low-salt buffers (containing 50 mM NaCl), the enzyme eluted in a single activity peak at an elution volume that appeared to correlate with a possible hexameric structure. In higher salt buffer (containing greater than 150 mM NaCl), the enzyme eluted as smaller assemblies (monomer/dimer). The recombinant helicase A was able to hydrolyze ATP or dATP with equal efficiency. The ATPase activity of the enzyme was absolutely DNA-dependent. The nucleotidase activities were comparable to those of the native enzyme. Kinetic analysis of the ATPase activity demonstrated that the Km of the enzyme was approximately 90 microM and the rate of ATP hydrolysis was approximately 20 ATP s-1 molecule-1. DNA sequences containing pyrimidine stretches were more effective activators than those containing purine stretches. However, poly(dC) appeared to be the most effective activator of the ATPase activity. The ATPase activity was inhibited by salt (NaCl) above 50 mM with a half-maximal inhibition at approximately 110 mM. It is likely that the active state of helicase A is hexameric. The helicase activity of the recombinant enzyme was stimulated significantly by the yeast replication protein A (RPA) and to a lower extent by the single-stranded DNA binding protein of E. coli (SSB). The DNA helicase migrated on a DNA template in a 5' --> 3' direction. Helicase A appeared to share a number of enzymatic characteristics including directionality, stimulation by RPA/SSB, and quaternary structure (monomer-hexamer) dynamics that are common to known replicative helicases such as DnaB helicase and the SV40 T-antigen. PMID- 9341219 TI - Differential poisoning of topoisomerases by menogaril and nogalamycin dictated by the minor groove-binding nogalose sugar. AB - The effect of DNA binding on poisoning of human DNA TOP1 has been studied using a pair of related anthracyclines which differ only by a nogalose sugar ring. We show that the nogalose sugar ring of nogalamycin, which binds to the minor groove of DNA, plays an important role in affecting topoisomerase-specific poisoning. Using purified mammalian topoisomerases, menogaril is shown to poison topoisomerase II but not topoisomerase I. By contrast, nogalamycin poisons topoisomerase I but not topoisomerase II. Consistent with the biochemical studies, CEM/VM-1 cells which express drug-resistant TOP2alpha are cross resistant to menogaril but not nogalamycin. The mechanism by which nogalamycin poisons topoisomerase I has been studied by analyzing a major topoisomerase I mediated DNA cleavage site induced by nogalamycin. This site is mapped to a sequence embedded in an AT-rich region with four scattered GC base pairs (bps) (at -10, -6, +2, and +12 positions). GC bps embedded in AT-rich regions are known to be essential for nogalamycin binding. Surprisingly, DNase I footprinting analysis of nogalamycin-DNA complexes has revealed a drug-free region from -2 to +9 encompassing the major cleavage site. Our results suggest that nogalamycin, in contrast to camptothecin, may stimulate TOP1 cleavage by binding to a site(s) distal to the site of cleavage. PMID- 9341220 TI - Klenow fragment-DNA interaction required for the incorporation of nucleotides opposite guanine and O6-methylguanine. AB - A mechanism by which the Klenow fragment of DNA polymerase I monitors the geometry of the base pairs may involve hydrogen bonds between the polymerase and the minor groove of the nascent base pair. The involvement of the 3-position of guanine in the template strand was examined by synthesizing oligodeoxynucleotides containing guanine and 3-deazaguanine and comparing the steady-state kinetics of the incorporation of all four dNTPs. The Vmax/Km decreased a significant amount (170-fold) only when dCTP was the co-substrate suggesting that a hydrogen bond exists only when the correct base pair is being replicated. This approach was also used to examine how the Klenow fragment interacts with the 3-position of the mutagenic base O6-methylguanine (O6mG). The Vmax/Km for the incorporation of dTTP opposite O6-methyl-3-deazaguanine (O6m3DG) was 1700-fold less than opposite O6mG. In contrast, a small 6-fold increase in Vmax/Km occurred for the incorporation of dCTP opposite O6m3DG relative to O6mG. This result suggests that the hydrogen bond between the Klenow fragment and O6mG is more important in the incorporation of dTTP opposite O6mG and may contribute to the mutagenicity of O6mG. PMID- 9341221 TI - Staphylococcal alpha-toxin: formation of the heptameric pore is partially cooperative and proceeds through multiple intermediate stages. AB - Staphylococcal alpha-toxin is a 293 residue polypeptide that assembles into pore forming heptamers, residues 118-140, thereby inserting to form an amphipathic beta-barrel in the lipid bilayer. Fluorometric analyses were here conducted using cysteine-substitution mutants site-specifically-labeled at positions 35 or 130 with the environmentally-sensitive fluorophore acrylodan. In conjunction with functional assays, three conformational states of the heptamer were defined, which may represent transitional configurations of the toxin molecule along its way to membrane insertion and pore formation. The first was the freshly assembled, SDS-sensitive heptamer alpha7*a, where a minor alteration in the environment of H35 with no change in the environment of the membrane-inserting stem domain was observed. In transition stage alpha7*b, the stem domain moved from a hydrophilic to a more hydrophobic environment, due to protein-protein interaction. Transition to alpha7*c involved a cooperative effect, in which residue 35 was forced by a neighboring molecule into a markedly hydrophobic environment. At this stage, the heptamers acquired SDS stability. The final pore conformation alpha7 resulted when the stem domain inserted into the lipid bilayer, an event that was driven by H35 within the respective protomer. A model thus evolved in which cooperative forces first lever H35 into a position that subsequently drives the pore-forming sequence within each respective protomer into the membrane. In accord with this model, when hybrid heptamers were formed between a functionally defective H35 substitution mutant and active toxin, only the latter inserted their pore-forming domain into the membrane. In a satisfying functional correlate, pores of reduced size were then generated. PMID- 9341222 TI - Effects of protein kinase A phosphorylation on signaling between cardiac troponin I and the N-terminal domain of cardiac troponin C. AB - During beta-adrenergic stimulation of the heart, there is a decrease in myofilament Ca2+ sensitivity mediated by the protein kinase A-(PKA-) induced phosphorylation of troponin I (cTnI). Phosphorylation, which occurs at Ser 23 and Ser 24 in an amino-terminal extension unique to cTnI, decreases the Ca2+ affinity of the amino-terminal regulatory site of cardiac troponin C (cTnC). In view of the antiparallel organization of the cTnI-cTnC complex [Krudy, G. A., Kleerekoper, Q., Guo, X., Howarth, J. W., Solaro, R. J., and Rosevear, P. R. (1994) J. Biol. Chem. 269, 23731-23735], it is not clear how the phosphorylation signal at one end of the complex affects the Ca2+ binding site at the other end. To address this question, we probed the interaction between cTnI and cTnC fragments, cTnC1-89 and cTnC90-162 (recombinant peptides corresponding to the N- and C-domains of cTnC). cTnI-Cys 5 mutant (S5C/C81I/C98S) and cTnC1-89 were fluorescently labeled with IAANS. When cTnI was phosphorylated, the affinity of Ca2+ for the cTnI-cTnC1-89 complex decreased significantly as indicated by a shift in the pCa50 value from 6.65 to 5.25. Upon phosphorylation, the affinity of cTnI for cTnC1-89 decreased by 3.8-fold in the absence of Ca2+ and 1.7-fold in the presence of Ca2+. In contrast to the case with full-length cTnC, neither cTnC1-89 nor cTnC90-162 induced significant structural changes in cTnI-Cys 5 as determined from intersite distance measurements between Cys 5 and Trp 192. Moreover, neither fragment of cTnC could significantly restore Ca2+ regulation of force generation, when exchanged into fiber bundles from which cTnC had been extracted. Our findings indicate that the transduction of PKA-induced phosphorylation signal from cTnI to the regulatory site of cTnC involves a global change in cTnI structure. PMID- 9341223 TI - Leucine 332 at the boundary between the fourth transmembrane segment and the cytoplasmic domain of Na+,K+-ATPase plays a pivotal role in the ion translocating conformational changes. AB - Mutants Gly330-->Ala, Leu332-->Ala, Leu332-->Pro, and Pro780-->Ala of the alpha1 isoform of rat kidney Na+,K+-ATPase were expressed in COS-1 cells to active site concentrations between 20 and 70 pmol per mg of membrane protein. The functional properties of the mutants were characterized by titrations of Na+-, K+-, and ATP dependencies of Na+,K+-ATPase activity as well as by a series of assays measuring the K+-dependence of the steady-state phosphoenzyme level, the kinetics of dephosphorylation under a variety of conditions, and the ADP-ATP exchange activity. In mutants Gly330-->Ala, Leu332-->Ala, and Leu332-->Pro, the molecular turnover number was reduced relative to that of the wild-type Na+,K+-ATPase, and the steady-state phosphoenzyme level was high even in the presence of several millimolar K+. At a low Na+ concentration in the absence of K+, mutants Leu332- >Pro and Gly330-->Ala displayed high ADP-ATP exchange activity and formed a high level of the ADP-sensitive phosphoenzyme (E1P). The phosphoenzyme decayed slowly following a jump in salt concentration and chase with ATP and K+. Hence, the conversion of E1P to the K+-sensitive phosphoenzyme (E2P) was inhibited in mutants Leu332-->Pro and Gly330-->Ala. In the Leu332-->Ala mutant, a high level of E2P was accumulated in the absence of K+, and the ADP-ATP exchange activity was low at low Na+ concentration in the absence of K+, but rose sharply on addition of K+. Dephosphorylation experiments indicated that in the Leu332-->Ala mutant K+ induced reversal of the phosphoenzyme interconversion, forming E1P from E2P. Leu332 is therefore a pivotal residue in the conformational change. Mutants Gly330-->Ala and Pro780-->Ala displayed reduced K+ affinities relative to the wild type, determined in three independent assays. PMID- 9341224 TI - Subtype-specific desensitization of human endothelin ETA and ETB receptors reflects differential receptor phosphorylation. AB - Endothelins regulate blood pressure in mammals through G protein-coupled receptors. Two receptor subtypes, ETA and ETB, exist which differ by their agonist profiles. Here we show subtype-specific differences in the inactivation of these endothelin receptors. Using a modified inositol phosphate accumulation assay, we found that stimulation of ETA by endothelin-1 results in sustained activation of the subtype, retaining >30% of its initial activity even 20 min after agonist administration, whereas the ETB rapidly deactivated after agonist stimulation, losing >80% of its initial activity within 5 min after endothelin application. The discrepancy in receptor inactivation is reflected by subtype specific differences in receptor phosphorylation. Whereas ETA failed to undergo ligand-induced phosphorylation, the ETB was rapidly phosphorylated in response to agonist stimulation. By contrast, the kinetics of ligand-induced internalization were essentially identical for the receptor subtypes, suggesting endothelin receptor internalization being independent of ligand-induced receptor phosphorylation. Interestingly, a strong correlation was observed between the time course of ETA inactivation and ETA internalization. Therefore, our data suggest a subtype-specific inactivation of human endothelin receptors: fast receptor phosphorylation in the case of ETB and slow receptor internalization in the case of ETA. Subtype-specific modulation of endothelin receptors may account for the short-term hypotensive effects of endothelins via rapidly down-regulating ETB receptors and the long-lasting hypertensive effects due to sustained ETA activation. PMID- 9341225 TI - Mapping of a binding site for ATP within the extracellular region of the Torpedo nicotinic acetylcholine receptor beta-subunit. AB - Using 2,8,5'-[3H]ATP as a direct photoaffinity label for membrane-bound nicotinic acetylcholine receptor (nAChR) from Torpedo marmorata, we have identified a binding site for ATP in the extracellular region of the beta-subunit of the receptor. Photolabeling was completely inhibited in the presence of saturating concentrations of nonradioactive ATP, whereas neither the purinoreceptor antagonists suramin, theophyllin, and caffeine nor the nAChR antagonists alpha bungarotoxin and d-tubocurarine affected the labeling reaction. Competitive and noncompetitive nicotinic agonists and Ca2+ increased the yield of the photoreaction by up to 50%, suggesting that the respective binding sites are allosterically linked with the ATP site. The dissociation constant KD of binding of ATP to the identified site on the nAChR was of the order of 10(-4) M. Sites of labeling were found in the sequence regions Leu11-Pro17 and Asp152-His163 of the nAChR beta-subunit. These regions may represent parts of a single binding site for ATP, which is discontinuously distributed within the primary structure of the N-terminal extracellular domain. The existence of an extracellular binding site for ATP confirms, on the molecular level, that this nucleotide can directly act on nicotinic receptors, as has been suggested from previous electrophysiological and biochemical studies. PMID- 9341226 TI - Distinct metal-thiolate clusters in the N-terminal domain of neuronal growth inhibitory factor. AB - Neuronal growth inhibitory factor (GIF), a brain-specific metallothionein-like protein (metallothionein-3), impairs the survival and neurite formation of cultured neurons. The metal distribution in isolated Cu4,Zn3-GIF is not known. In the present studies, the metal-thiolate clusters formed with monovalent and divalent metal ions in the N-terminal domain of human GIF [GIF(1-32)] were investigated. The cluster formation was followed by using electronic absorption, circular dichroism (CD), and magnetic circular dichroism (MCD), and in the case of Cu(I) complexes also by luminescence spectroscopy at 77 K. With Cu(I) ions, two well-defined clusters are formed involving the nine cysteine ligands of GIF(1 32), i.e., Cu4S9- and Cu6S9-clusters. In contrast to the Cu6S9-cluster, the Cu4S9 cluster shows a remarkable stability to air oxidation. As similar properties and spectral features have also been observed with isolated Cu4-5,Zn2-3-GIF, the presence of a Cu4-cluster in this GIF form is suggested. The studies with Zn(II), Cd(II), and Co(II) ions indicated the presence of a Me3S9-cluster in GIF(1-32). However, spectral features of these metal derivatives substantially differ from those reported for the corresponding Me3S9-cluster in the beta-domain of metallothioneins, suggesting structural differences. A large conformational flexibility of the Zn3- and Cd3-GIF(1-32) structures, characterized by short T2 proton relaxations, precluded their investigation by NMR methods. The significance of Cu- and Zn-clusters for the structure of biologically active GIF(1-32) is discussed. PMID- 9341227 TI - Roles for the two cysteine residues of AhpC in catalysis of peroxide reduction by alkyl hydroperoxide reductase from Salmonella typhimurium. AB - The catalytic properties of cysteine residues Cys46 and Cys165, which form intersubunit disulfide bonds in the peroxidatic AhpC protein of the alkyl hydroperoxide reductase (AhpR) system from Salmonella typhimurium, have been investigated. The AhpR system, composed of AhpC and a flavoprotein reductase, AhpF, catalyzes the pyridine nucleotide-dependent reduction of organic hydroperoxides and hydrogen peroxide. Amino acid sequence analysis of the disulfide-containing tryptic peptide demonstrated the presence of two identical disulfide bonds per dimer of oxidized AhpC located between Cys46 on one subunit and Cys165 on the other. Mutant AhpC proteins containing only one (C46S and C165S) or no (C46,165S) cysteine residues were purified and shown by circular dichroism studies to exhibit no major disruptions in secondary structure. In NADH dependent peroxidase assays in the presence of AhpF, the C165S mutant was fully active in comparison with wild-type AhpC, while C46S and C46,165S displayed no peroxidatic activity. In addition, only C165S was oxidized by 1 equiv of hydrogen peroxide, giving a species that was stoichiometrically reducible by NADH in the presence of a catalytic amount of AhpF. Oxidized C165S also reacted rapidly with a stoichiometric amount of the thiol-containing reagent 2-nitro-5-thiobenzoic acid to generate a mixed disulfide, and was susceptible to inactivation by hydrogen peroxide, strongly supporting its identification as a cysteine sulfenic acid (Cys46-SOH). The lack of reactivity of the C46S mutant toward peroxides was not a result of inaccessibility of the remaining thiol as demonstrated by its modification with 5, 5'-dithiobis(2-nitrobenzoic acid), but could be due to the lack of a proximal active-site base which would support catalysis through proton donation to the poor RO- leaving group. Our results clearly identify Cys46 as the peroxidatic center of AhpC and Cys165 as an important residue for preserving the activity of wild-type AhpC by reacting with the nascent sulfenic acid of the oxidized protein (Cys46-SOH) to generate a stable disulfide bond, thus preventing further oxidation of Cys46-SOH by substrate. PMID- 9341228 TI - Requirement for the two AhpF cystine disulfide centers in catalysis of peroxide reduction by alkyl hydroperoxide reductase. AB - AhpF, the alkyl hydroperoxide reductase component which transfers electrons from pyridine nucleotides to the peroxidase protein, AhpC, possesses two redox-active disulfide centers in addition to one FAD per subunit; the primary goal of these studies has been to test for the requirement of one or both of these disulfide centers in catalysis. Two half-cystine residues of one center (Cys345Cys348) align with those of the homologous Escherichia coli thioredoxin reductase (TrR) sequence (Cys135Cys138), while the other two (Cys129Cys132) reside in the additional N-terminal region of AhpF which has no counterpart in TrR. We have employed site-directed mutagenesis techniques to generate four mutants of AhpF, including one which removes the N-terminal disulfide (Ser129Ser132) and three which perturb the TrR-like disulfide center (Ser345Ser348, Ser345Cys348, and Cys345Ser348). Fluorescence, absorbance, and circular dichroism spectra show relatively small perturbations for mutations at the disulfide center proximal to the flavin (Cys345Cys348) and no changes for the Ser129Ser132 mutant; identical circular dichroism spectra in the ultraviolet region indicate unchanged secondary structures in all mutants studied. Oxidase and transhydrogenase activities are preserved in all mutants, indicating no role for cystine redox centers in these activities. Both DTNB and AhpC reduction by AhpF are dramatically affected by each of these mutations, dropping to less than 5% for DTNB reductase activity and to less than 2% for peroxidase activity in the presence of AhpC. Reductive titrations confirm the absence of one redox center in each mutant; even in the absence of Cys345Cys348, the N-terminal redox center can be reduced, although only slowly. These results emphasize the necessity for both redox-active disulfide centers in AhpF for catalysis of disulfide reductase activity and support a direct role for Cys129Cys132 in mediating electron transfer between Cys345Cys348 and the AhpC active-site disulfide. PMID- 9341229 TI - The roles of conserved carboxylate residues in IMP dehydrogenase and identification of a transition state analog. AB - IMP dehydrogenase (IMPDH) catalyzes the oxidation of IMP to XMP with the concomitant reduction of NAD+; the enzyme is activated by K+. This reaction is the rate-limiting step in de novo guanine nucleotide biosynthesis. In order to identify functionally important residues in IMPDH, including those involved in substrate and K+ binding, we have mutated 11 conserved Asp and Glu residues to Ala in Escherichia coli IMPDH. The values of kcat, Km, and Ki for GMP, XMP, mizoribine 5'-monophosphate (MMP), and beta-methylene-tiazofurin adenine dinucleotide (TAD) were determined. Five of these mutations caused a significant change (>/=10-fold) in one of these parameters. The Asp248 --> Ala mutation caused 100-fold decrease in the value of kcat and a 25-fold increase in the value of Kii for TAD; these observations suggest that Asp248 is in the NAD+ binding site. The Asp338 --> Ala mutation caused a 600-fold decrease in the value of kcat, but only a 5-10-fold increase in the values of Km for IMP and Kis for IMP analogs, suggesting that Asp338 may be involved in acid-base catalysis as well as IMP binding. The remaining three residues, Asp13, Asp50, and Glu469, appear to be involved in K+ activation; these residues may be ligands at one or more K+ binding sites. Interestingly, changes in the values of Ki for MMP correlate with changes in kcat/KmKm of IMPDH, while no such correlation is observed for GMP, XMP, and TAD. This observation indicates that MMP is a transition state analog for the IMPDH reaction. PMID- 9341231 TI - The paradigm that all oxygen-respiring eukaryotes have cytosolic CuZn-superoxide dismutase and that Mn-superoxide dismutase is localized to the mitochondria does not apply to a large group of marine arthropods. AB - The enzyme superoxide dismutase (SOD), which catalyzes the dismutation of the superoxide radical, is present in the cytosol and mitochondria of all oxygen respiring eukaryotes. The cytosolic form contains copper and zinc (CuZnSOD), whereas the mitochondrial form contains manganese (MnSOD). The latter protein is synthesized in the cytosol as a MnSOD precursor, containing an N-terminal mitochondrial-targeting sequence. CuZnSOD is sensitive toward cyanide (CN) and hydrogen peroxide (H2O2), but MnSOD is not. Assays for SOD activity in cytosol from the hepatopancreas of the blue crab, Callinectes sapidus, showed the presence of a CN/H2O2-insensitive form of SOD. No CN/H2O2-sensitive CuZnSOD was found. This unexpected phenomenon was shown to occur in all decapod crustacea (crabs, lobsters, shrimp) examined. The cytosolic and mitochondrial SODs of C. sapidus were purified by means of ion-exchange, size-exclusion, and reverse-phase HPLC. The cytosolic SOD is a homodimeric protein, which exists in a monomer-dimer equilibrium (24 kDa left and right arrow 48 kDa). The protein contains approximately 1 Mn per subunit. No copper or zinc is present. Amino acid sequence analysis identified the novel cytosolic SOD as a MnSOD precursor with an abnormal mitochondrial-targeting sequence. The mitochondrial SOD of C. sapidus is similar to the MnSOD found in other eukaryotes. N-Terminal amino sequences of mitochondrial and cytosolic blue crab MnSOD differ in several positions. The MnSODs are thus encoded for by two different genes. The paradigm that all eukaryotes contain intracellular CuZnSOD and that MnSOD occurs exclusively in the mitochondria appears not to apply to a large group of marine arthropods. PMID- 9341230 TI - Assembly of a [2Fe-2S]2+ cluster in a molecular variant of Clostridium pasteurianum rubredoxin. AB - The rubredoxin from Clostridium pasteurianum contains a single iron atom bound to the polypeptide chain by cysteines 6, 9, 39, and 42. The C42A variant of this protein has been prepared by site-directed mutagenesis and heterologous expression of the gene in Escherichia coli. The mutated protein was found to contain an unexpected chromophore that has been characterized by a variety of techniques. UV-visible absorption and resonance Raman spectra were strongly reminiscent of those of [2Fe-2S] proteins. Mossbauer spectra of the oxidized chromophore isolated in oxygen-free conditions indicated low-temperature diamagnetism resulting from antiferromagnetically coupled high-spin ferric ions. Analysis of X-ray absorption fine structure spectra yielded an Fe-Fe distance of 2.68 A. Colorimetric assays of iron and inorganic sulfide showed that the two elements are present in a 1:1 ratio. Electrospray-ionization mass spectra displayed a major component at M = 6190 Da, i.e. the molecular mass of the C42A apoprotein plus two atomic masses of iron and two atomic masses of sulfur. Taken together, these data show that a mere point mutation allows the stabilization of a binuclear [2Fe-2S] cluster in a protein that normally accommodates a mononuclear Fe(Scys)4 site. Assembly of a [2Fe-2S] cluster may occur because rubredoxin assumes a similar fold around its metal center as the [2Fe-2S] Rieske protein. Alternatively, a more extensive structural rearrangement of the polypeptide chain of the C42A rubredoxin variant may be considered as well. PMID- 9341232 TI - Raman spectroscopic and light-induced kinetic characterization of a recombinant phytochrome of the cyanobacterium Synechocystis. AB - A phytochrome-encoding cDNA from the cyanobacterium Synechocystis has been heterologously expressed in Escherichia coli and reconstituted into functional chromoproteins by incubation with either phycocyanobilin (PCB) or phytochromobilin (PPhiB). These materials were studied by Raman spectroscopy and nanosecond flash photolysis. The Raman spectra suggest far-reaching similarities in chromophore configuration and conformation between the Pfr forms of Synechocystis phytochrome and the plant phytochromes (e.g. phyA from oat), but some differences, such as torsions around methine bridges and in hydrogen bonding interactions, in the Pr state. Synechocystis phytochrome (PCB) undergoes a multistep photoconversion reminiscent of the phyA Pr --> Pfr transformation but with different kinetics. The first process resolved is the decay of an intermediate with red-shifted absorption (relative to parent state) and a 25 micros lifetime. The next observable intermediate grows in with 300 (+/-25) micros and decays with 6-8 ms. The final state (Pfr) is formed biexponentially (450 ms, 1 s). When reconstituted with PPhiB, the first decay of this Synechocystis phytochrome is biexponential (5 and 25 micros). The growth of the second intermediate is slower (750 micros) than that in the PCB adduct whereas the decays of both species are similar. The formation of the Pfr form required fitting with three components (350 ms, 2.5 s, and 11 s). H/D Exchange in Synechocystis phytochrome (PCB) delays, by an isotope effect of 2.7, both growth (300 micros) and decay rates (6-8 ms) of the second intermediate. This effect is larger than values determined for phyA (ca. 1.2) and is characteristic of a rate limiting proton transfer. The formation of the Pfr state of the PCB adduct of Synechocystis phytochrome shows a deuterium effect similar as phyA (ca. 1.2). Activation energies of the second intermediate in the range 0-18 degrees C are 44 (in H2O/buffer) and 48 kJ mol-1 (D2O), with essentially identical pre-exponential factors. PMID- 9341233 TI - Acquisition of native beta-strand topology during the rapid collapse phase of protein folding. AB - The 98 residue C-terminal domain of the cell-surface receptor protein CD2 (CD2.D1) has a beta-sandwich fold belonging to the immunoglobulin superfamily but lacking the usual disulfide bridges. Kinetic studies on the folding/unfolding of CD2.D1 reveal that folding proceeds through a rapidly formed intermediate state [Parker, M. J., & Clarke, A. R. (1997) Biochemistry 36, 5786-5794]. To characterize the structural properties of this intermediate we have performed a series of amide hydrogen exchange studies using the pH competition method, in which folding and exchange are initiated simultaneously. The complex beta-sheet topology of this molecule makes it an ideal object for examining the acquisition of backbone hydrogen bonds made between sequence-local and sequence-distant segments of the chain during folding. The pattern of protected amides in the intermediate reveal that the essential features of the beta-sheet topology of CD2.D1 are defined early in the folding pathway, before the development of intimate side chain interactions characteristic of the native state. The results are discussed in light of current issues concerning the mechanistic relevance of kinetic protein folding intermediates. PMID- 9341235 TI - Magnetic resonance studies on the active site and metal centers of Bradyrhizobium japonicum porphobilinogen synthase. AB - Porphobilinogen synthase (PBGS) is a metalloenzyme which catalyzes the asymmetric condensation of two molecules of 5-aminolevulinic acid (ALA) to form porphobilinogen. There are at least four types of PBGS, categorized according to metal ion usage. The PBGS from Bradyrhizobium japonicum requires Mg(II) in catalytic metal site A, has an allosteric Mg(II) in metal site C, and also contains an activating monovalent cation binding site [Petrovich et al. (1996) J. Biol. Chem. 271, 8692-8699]. 13C NMR and Mn(II) EPR have been used to probe the active site and Mg(II) binding sites of this 310 000 dalton protein. The 13C NMR chemical shifts of enzyme-bound product demonstrate that the chemical environment of porphobilinogen bound to B. japonicum PBGS is different from that of PBGS which contains Zn(II) rather than Mg(II) at the active site. Use of Mn(II) in place of Mg(II) broadens the NMR resonances of enzyme-bound porphobilinogen, providing evidence for a direct interaction between MnA and product at the active site. Prior characterization of the enzyme defined conditions in which the divalent cation occupies either the A or the C site. Mimicking these conditions allows Mn(II) EPR observation of either MnC or MnA. The EPR spectrum of MnC is significantly broader and less intense than "free" Mn(II), but relatively featureless. The EPR spectrum of MnA is broader still and more asymmetric than MnC. The EPR data indicate that the coordination spheres of the two metals are different. PMID- 9341234 TI - Stopped-flow kinetic investigations of conformational changes of pig kidney Na+,K+-ATPase. AB - The kinetics of Na+-dependent partial reactions of the Na+,K+-ATPase were investigated via the stopped-flow technique using the fluorescent labels RH421 and BIPM. After the enzyme is mixed with MgATP, both labels give almost identical kinetic responses. Under the chosen experimental conditions two exponential time functions are necessary to fit the data. The dominant fast phase, 1/tau1 approximately 180 s-1 (saturating [ATP] and [Na+], pH 7.4 and 24 degrees C), is attributed to phosphorylation of the enzyme and a subsequent conformational change (E1ATP(Na+)3 --> E2P(Na+)3 + ADP). The rate of the phosphorylation reaction measured by the acid quenched-flow technique was 190 s-1 at 100 microM ATP, suggesting that phosphorylation controls the kinetics of the RH421 signal and that the conformational change is very fast (>/=600 s-1). The rate of the RH421 signal was optimal at pH 7.5. The Na+ concentration dependence of 1/tau1 showed half-saturation at a Na+ concentration of 8-10 mM with positive cooperativity involved in the occupation of the Na+ binding sites. The apparent dissociation constant of the high affinity ATP binding site determined from the ATP concentration dependence of 1/tau1 was 7.0 (+/-0.6) microM, while the apparent Kd for the low affinity site and the rate constant for the E2 to E1 conformational change evaluated in the absence of Mg2+ were 143 (+/-17) microM and 2MeOGal > 2FGal congruent with Gal > 2HGal, which suggests that both polar and nonpolar residues surround the C2 locus of galactose, consistent with the observed high affinity of WBA I toward GalNAc where the acetamido group at C2 position is probably stabilized by both nonpolar interactions with the methyl group and polar interactions with the carbonyl group. The binding of C6 derivatives follows the order Gal > 6FGal > D-Fuc >> 6MeOGal congruent with L-Ara, indicating the presence of favourable polar interactions with a hydrogen bond donor in the vicinity. On the basis of these results the hydrogen bond donor-acceptor relationship of the complexation of methyl-alpha-D-galactopyranoside with the primary combining site of WBA I is proposed. PMID- 9341237 TI - Structure and function of Cerebratulus lacteus neurotoxin B-IV: tryptophan-30 is critical for function while lysines-18, -19, -29, and -33 are not required. AB - The Cerebratulus lacteus B-toxins are a family of polypeptide neurotoxins known to bind to crustacean voltage-sensitive sodium channels. We have previously shown that in the most abundant homolog, toxin B-IV, Arg-17 in the N-terminal helix and a positive charge at position 25 in the loop region are essential for function. In this report, we target a tryptophan residue at position 30, as well as lysine residues found in both the N-terminal helix and loop regions by polymerase chain reaction mutagenesis, to determine their contributions to toxin activity. Substitution of Trp-30 with a serine causes a more than 40-fold reduction in specific toxicity, whereas replacement by tyrosine and phenylalanine is well tolerated. The secondary structures of both these muteins are identical to that of the wild-type toxin as determined by circular dichroism spectroscopy. Thermal denaturation experiments also show that their conformational stabilities are intact. These results demonstrate that an aromatic residue at this position is required for toxin function. Charge neutralizing substitutions of Lys-18 and Lys 19 located in the N-terminal helix have very little effect on toxicity, suggesting the nonessentiality of these residues. Similar results are also obtained for the charge neutralizing muteins for Lys-29 and Lys-33 in the loop region. Interestingly, reduction experiments demonstrate that both K29N and W30S are more sensitive to reducing agent than wild-type B-IV, raising the possibility that the loop sequence may modulate toxin stability. PMID- 9341239 TI - The neural integrators of the mammalian saccadic system. AB - The neural velocity to position integrators transform the saccade related signal of the burst generators into an eye position related tonic signal they convey to motoneurons. They are largely confined to three heavily interconnected midbrain structures: 1) The interstitial nucleus of Cajal (NIC), 2) The nucleus prepositus hypoglossi (NPH), 3) The vestibular nuclei (VN). Integration in the horizontal and vertical planes is accomplished largely independently by the NPH-VN and the NIC-VN complexes, respectively. Cells in these regions carry a more or less intense phasic signal related to saccades and a tonic signal related to eye position. Depending on the relationship between the rate of their discharge and the position of the eyes, these cells have been further subdivided into regular or irregular, more or less sensitive, and bi-directionally or uni-directionally modulated. The present review provides a brief description of their discharge pattern and that of burst neurons and extraocular motoneurons. Then, evidence concerning the input-output connections of relevant cell classes is summarized. Finally, several modelling attempts to simulate the neural velocity-to-position integrators are presented and their verisimilitude is evaluated in the light of psychophysical, anatomical, physiological and neurological evidence. PMID- 9341238 TI - Methylation of the Escherichia coli chemotaxis receptors: intra- and interdimer mechanisms. AB - The mechanism(s) of methylation of the Escherichia coli chemotaxis receptors was analyzed by experiments involving the construction of a series of aspartate receptor variants. Truncation of five or more residues from the C-terminal end of the aspartate receptor, which prevents the methyltransferase from binding to the receptor, resulted in very low rates of methylation, indicating that the methyltransferase is activated by binding to the receptor. Coexpression of a receptor variant that is unmethylatable but able to C-terminally bind the methyltransferase resulted in much higher methylation rates for all of the truncated receptors. By preventing the possibility of subunit exchange between receptor variants, we showed that the truncated receptors were methylated via an interdimer mechanism. The interdimer methylation rates of the truncated receptors were found to be 3-fold lower than the methylation rate of the unaltered receptor, suggesting that intradimer methylation as well as interdimer methylation accounts for the methylation of the unaltered receptor. In addition, the presence of the cytoplasmic signaling proteins, which have been shown to cause receptor clustering, did not influence the rates of methylation. PMID- 9341240 TI - Cellular mechanisms of feline immunodeficiency virus (FIV)-induced neuropathogenesis. AB - The high incidence of neurologic dysfunction from human immunodeficiency virus (HIV) infection has heightened interest in neuropathogenesis of other lentiviruses, including that associated with feline immunodeficiency virus (FIV). Both HIV and FIV efficiently enter the central nervous system and cause primary neurological disease that is not attributable to opportunistic infections or systemic disease. Cells in the brain infected by FIV are similar to those observed in HIV infection, both viruses infect macrophages, microglia, and astrocytes. Although substantial neuronal loss can occur in the cortex of HIV- or FIV-infected patients, most studies agree that neurons are not infected and indirect mechanisms of neurotoxicity are postulated. This review describes recent information on the neuropathogenesis of FIV and how this information correlates with what is known about the neuropathogenesis of HIV. Although the pathogenesis of neurological dysfunction in HIV- and FIV-infected patients is far from clear, it is becoming increasingly evident that the relationship between lentivirus presence in the brain and neurological signs is not straightforward and cannot be explained by simple cytolytic infection. The observed neurologic dysfunction is likely multifactorial and complex involving an intricate web of subcellular pathways and neurotoxic factors interacting with multiple cell types. PMID- 9341241 TI - The future of associate degree nursing. PMID- 9341242 TI - The future of associate degree nursing. PMID- 9341243 TI - Isn't it time? PMID- 9341244 TI - Stay informed: informational resources for the community health nurse. PMID- 9341245 TI - Nursing creativity. PMID- 9341246 TI - NLN challenges nursing schools nationwide. PMID- 9341247 TI - Helping at-home caregivers ask for help. PMID- 9341248 TI - Reauthorization '97. The calm before the storm. Reauthorization of Higher Education Act of 1965. PMID- 9341250 TI - NLN president's acceptance speech. PMID- 9341249 TI - Life-terminating choices. A framework for nursing decision making. AB - During the 1990s, assisted suicide has become the subject of public debate and legislative action across the nation. To help nurses explore the issues involved, an NLN Task Force prepared this report. PMID- 9341251 TI - Passover. PMID- 9341252 TI - The Jagged2 gene maps to chromosome 12 and is a candidate for the lgl and sm mutations. PMID- 9341253 TI - [Significance of arterial ketone body ratio as a parameter of oxygen metabolism in cardiac surgery]. AB - Postoperative course after cardiac surgery is characterized by a progressive increased cellular oxygen demand and limited oxygen supply. It is mandatory to assess the adequacy of tissue oxygenation and to correct inadequate oxygenation rapidly in cardiac surgery. The present study was designed to evaluate the relationship between the arterial ketone body ratio (AKBR) and the status of oxygen demand and supply relationship in cardiac surgery. We measured oxygen consumption, oxygen delivery, oxygen extraction, mixed venous oxygen saturation, lactate, lactate/pyruvate (L/P) and AKBR in 43 patients undergoing open heart surgery at selected 10 periods before, during and after cardiopulmonary bypass (CPB). AKBR significantly decreased immediately after the beginning of CPB and returned to pre-CPB level more quickly than lactate and L/P did. AKBR at several periods a significant correlation with lactate and L/P at delayed periods. There was a significant correlation between AKBR and L/P during and after CPB. Furthermore, oxygen consumption, oxygen extraction, mixed venous oxygen saturation, and lactate were correlated with AKBR significantly. But there was no correlation between the parameter of oxygen metabolism and both lactate and L/P. In conclusion, it appears from these data that AKBR in cardiac surgery may be helpful as a rapid guide for estimating the degree of anaerobiosis. PMID- 9341254 TI - [Size estimation method for patch used in reconstruction of LV cavity]. AB - The reconstruction of LV cavity is accomplished by suturing a patch to the viable myocardium to exclude the infarcted area from the high LV pressures. However, there is no clear guideline to estimate the size of patch used for LV reconstruction. We have designed a new method to determine the correct patch size, and applied it in 5 cases. The suture line of the patch is at the junction of contractile (functional) and infarcted portions of LV. The patch size is determined by the length of AB, termed "a", as the base, where "point A" represents the junction on the LV anterior wall side, and "point B" the junction of the LV posterior wall side, from RAO 30 degrees projection of the left ventriculogram obtained by cardiac catheterization. In LV aneurysm, we designed the patch in the range of a/2 < l < or = pi a/2, where patch length on RAO 30 degrees is considered "l". An effort was made to reconstruct to normalize LV volume and contour by designing the patch size to be a/2 < l < a, particularly when the contractile portion was enlarged by aneurysm. On the other hand, in post AMI VSD, LV contractile portion is not enlarged in early stage. Therefore, the patch was designed in the range of a < l < or = pi a/2 to maintain LV volume. Postoperative LV volume can be calculated prior to surgery, by using the lengths of the designed patch. Postoperative analysis indicated that the actual LV volume and contour were almost identical to our estimation. This method is very useful in planning the patch size for LV reconstruction. PMID- 9341255 TI - [Clinical results of single-stage mobilization of pectoral muscle flaps and omental transposition for infected mediastinitis after open heart surgery]. AB - The purpose of this study was to retrospectively evaluate the outcome of refractory infected mediastinitis managed primarily with mobilization of pectoral muscle flaps and omental transposition. From January 1992 to December 1995, infected mediastinitis occurred in 11 (2.5%) of 447 consecutive patients. All patients required sternal debridement. The wound was thoroughly irrigated with a solution of 0.5% povidone-iodine in physiological saline after debridement and then the defect was repaired. Reconstruction of the chest wall was attained using pectoral muscle flaps in seven patients and pectoral muscle flaps and omental transposition in four. Antibiotic therapy was provided for 6 weeks or more according to the regimen in North America. No hospital deaths occurred after surgery. Significant early complications occurred in four patients. The reasons for the prolonged hospitalization were a recurrent wound infection, prosthetic valve endocarditis and saphenous vein graft pseudoaneurysm formation caused by Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-resistant Staphylococcus epidermidis (MRSE). Length of stay in ICU after surgical treatment was range 1 to 140 days (an average of 11 +/- 3 days in 9 patients without complications in ICU). Duration between surgical treatment and discharge was range 47 to 300 days (an average of 58 +/- 8 days in 7 patients without significant early complications). At the time of this report, the patients are doing well with no signs of recurrence of infection. The mean follow-up was 28.8 months (range 8 to 48 months). We conclude that single-stage mobilization of pectoral muscle flaps together with omental transposition is very usefull for managing refractory infected mediastinitis. But careful follow-up is needed after this procedure in case of MRSA-caused mediastinitis because of its tendency to recur. PMID- 9341257 TI - [Analysis of small lung nodules diagnosed by open lung or thoracoscopic biopsy]. AB - We examined clinicopathologic findings in 86 cases with peripheral lung nodules less than 30 mm in size diagnosed by open lung or video-assisted thoracoscopic surgery (VATS) biopsy. Biopsies were conducted because of the new appearance or enlargement of nodules as evidenced in a comparison with retrospective chest films in 47 patients, X-ray findings of malignancy suspicion without retrospective films in 13, enlargement of nodules after the administration of antituberculosis agents in 9, and a past history of malignancy in 17. Mean tumor size was 18.1 mm in primary lung cancer (n = 29), 16.2 mm in metastatic lung cancer (n = 13), 16.3 mm in tuberculosis (n = 18), 15.3 mm in nonspecific inflammation (n = 12), 16.7 mm in benign lung tumors (n = 7), 7.5 mm in intrapulmonary lymph node (n = 2), and 19.4 mm in others (n = 5). Among primary lung cancers with a clear N-factor, the percentage of T1N0M0 cancers was up to 72%. No significant difference was observed in either of the reasons for these biopsies and the size of nodules among diseases. To detect early lung cancer and increase the rate of cure, small pulmonary nodules that could be hardly diagnosed using bronchoscopic or needle aspiration biopsy should be diagnosed positively using VATS biopsy. PMID- 9341256 TI - [Contralateral pneumothorax after lung resection]. AB - Nine hundred and seventy-three consecutive patients were referred to our hospital for thoracotomy to treat chest diseases between January 1, 1981, and December 31, 1995. Of these patients, 20 males were readmitted within a mean of 20 months with a diagnosis of contralateral pneumothorax. Sixteen of the patients with a mean age of 28.5 years (range 16-76 years of age) had been operated on for bullous lung disease. The remaining four, with a mean age of 60.8 years (range 54-71), had been operated on for lung cancer. All of the 20 patients had received unilateral thoracotomy for lung resection. One patient had undergone pneumonectomy for lung cancer; three had undergone lobectomy; and 16 had been treated by partial lung resection. The patient who had undergone pneumonectomy was found to have contralateral pulmonary metastasis of lung cancer. In the other 19 patients, emphysematous bulla was the origin of the contralateral pneumothorax. The mean value of body mass index (BMI) of the group was 18.4 as compared to 21.7 in the patients who did not go on to develop contralateral pneumothorax, a significant difference (p < 0.05). In conclusion, postoperative contralateral pneumothorax was correlated to the existence of emphysematous changes of the lung and a significantly lower BMI. We conclude that patients with BMIs less than 20 may be at increased risk of developing postoperative contralateral pneumothorax. PMID- 9341258 TI - [A case of recurrent bronchogenic cyst 15 years after initial operation]. AB - There are few reports on the postoperative recurrence of bronchogenic cysts. We conducted a re-operation on a 57-year-old man with a bronchogenic cyst 15 years after an initial operation. His history showed an earlier operation for a bronchogenic cyst at the age of 42 at another hospital. 15 years after this initial operation, he suffered from common cold like symptoms, and was referred to our hospital, because of an abnormal shadow on his chest X-ray. A chest CT and MRI revealed an oval tumor just under the right intermediate bronchus. We suspected it was a recurrence of the bronchogenic cyst, and an operation was performed. The cyst was firmly adhered to the lung, and at the upper site of the cyst, a region adjacent to the intermediate bronchus was adhered to the bronchial wall. The histological findings were similar to those of 15 years previously. The cyst wall lined with pseudostratified columnar ciliary epithelium with muscular layer, which led to a diagnosis of a bronchogenic cyst. Congenital cysts, including bronchogenic cysts, are considered to originate in abnormal primordia. If there are remaining abnormal primordia, a recurrence of the disease can occur. PMID- 9341259 TI - [An experience of the modified Norwood's operation for hypoplastic left heart syndrome with aberrant origin of right subclavian artery and persistent left superior vena cava--the procedure without total circulatory arrest and cardiac arrest]. AB - We reported a successful case of the modified Norwood operation for a 21-day-old neonate with hypoplastic left heart syndrome (MS and AS) associated with an aberrant right subclavian artery and a persistent left superior vena cava. The modified Norwood operation was performed without total circulatory arrest and Cardiac arrest. A 4 mm Gore-Tex graft, which was anastomosed between the right carotid artery and the right pulmonary artery for systemic-pulmonary shunt, was used for cerebral perfusion during aortic arch reconstruction. Coronary perfusion was performed with a small cannula placed on the relatively large ascending aorta during anastomosis between the main pulmonary artery and the ascending aorta. Equine pericardial patch was used for aortic arch reconstruction and the ascending aorta was directly anastomosed to a part of the main PA. Postoperative course was uneventful and postoperative MRI revealed no stenosis of the aortic arch and the pulmonary artery. PMID- 9341260 TI - [A report of two cases of Carpentier-Edwards pericardial mitral valve malfunction due to neointimal overgrowth expanding to valve cusps]. AB - Two cases of Carpentier-Edwards pericardial mitral valve malfunction due to neointimal overgrowth were reported. Case one was a 51-year-old female undergone redo mitral valve replacement at nine years after first operation. Removed valve showed remarkable overgrowth of neointima expanding to the valve cusps. Case two was a 67-year-old male. A valve removed at nine years after first operation and at 1.5 years after recovery of prosthetic valve endocarditis. Removed valve also showed neointimal overgrowth expanding to the valve cusps. Although we experienced only two cases of neointimal overgrowth, these findings were considered being important in durability of Carpentier-Edwards pericardial valve. PMID- 9341261 TI - [A case report of thoracic aortic aneurysm associated with tubular hypoplasia]. AB - A 55-year-old female was diagnosed as thoracic aortic aneurysm associated with tubular hypoplasia. A saccular aneurysm occupied the aortic arch between left common carotid and left subclavian arteries, arising from the cranial wall of the tubular hypoplasia. The aneurysmectomy was performed under cardiopulmonary bypass with selective cerebral perfusion and woven Dacron graft was implanted. The post operative course was uneventful and she was discharged 32 days after the operation. Aneurysm associated with coarctation usually is thin walled, therefore an early surgical treatment should be urged. PMID- 9341262 TI - [A surgical case of aorto-pulmonary septal defect in a low weight neonate]. AB - We report a surgical case of aorto-pulmonary septal defect (APSD) in a neonate weighing 1693 gm. A male twin baby delivered after 39 weeks and 5 days of gestational period was diagnosed as APSD. RAA, PFO and PLSVC by a echocardiography and a MRI. Because of the progressive cardiac failure, operation was performed under cardiopulmonary bypass and profound hypothermic circulatory arrest at 30 days of age and weighing 1693 gm. APSD was closed completely by a Dacron patch. Postoperative course was almost uneventful except for pulmonary hypertension crisis. He recovered without brain damage. PMID- 9341263 TI - [A resected case of diffuse malignant pleural mesothelioma diagnosed by thoracoscopic biopsy]. AB - A 53-year-old male was admitted with cough and chest pain. A chest X-ray film showed left pleural effusion and a chest CT revealed irregular thickening of the pleura. Pleural fluid cytology and percutaneous needle biopsy were negative for malignancy. Thoracoscopic findings revealed fibrin network with pleural effusions and yellow-white pleural thickening, but neither nodules nor masses were found. The thoracoscopic biopsy specimen from the pleural thickening resulted in the diagnosis of malignant pleural mesothelioma. Left pleuropneumonectomy with mediastinal lymph node dissection was performed. Since detailed inspection of the pleural cavity and taking large biopsy samples under thoracoscopic examination are possible, we consider thoracoscopic biopsy to be a useful method for obtaining diagnosis of malignant pleural mesothelioma. Pleuropneumonectomy and systematic lymph node dissection of the pulmonary hilum and mediastinum were believed to be necessary for the surgical treatments. PMID- 9341264 TI - [A case of hemodialysis-associated innominate vein stenosis inducing superior vena cava syndrome]. AB - Twenty years after making an arteriovenous shunt in the left arm, a 45-year-old man on hemodialysis developed progressive swelling in the face to left arm and venous dilatation on the left anterior chest. Venogram disclosed severe stenosis of the left innominate vein at the junction of the superior vena cava, which was considered to be a primary lesion because he had no history of subclavian vein cannulation or mediastinal disease. Surgical resection of the stenotic lesion and direct anastomosis of the innominate vein resulted in a rapid recovery of the symptom of venous hypertension. This is a rare case of hemodialysis-associated large vein complication leading to superior vena cava syndrome. PMID- 9341265 TI - [A case of CABG under the cardiac arrest induced by a short acting beta-blocker without clamping the aorta]. AB - A seventy one year old woman had a coronary artery bypass grafting. No touch technique to her ascending aorta was applied due to the severely atherosclerotic aorta. Cardiac arrest was induced by a large dose of short acting beta-blocker (Esmolol) without cross-clamping the aorta under the normothermic cardio pulmonary bypass. The heart was flaccid and rotated easily while the coronary anastomoses were performed. Both of the internal thoracic arteries were grafted individually to the anterior descending artery and to the circumflex artery. The cardiac beats were resumed with the ordinary inotropic support and then the cardio-pulmonary bypass was weaned off. Her postoperative course was uneventful and the cardiac enzyme level was not elevated. Both of the grafts were revealed patent by the postoperative angiography. Esmolol had played an important roll to perform excellent anastomoses and to protect the myocardium. It was concluded that this technique could be one of the suitable modality for patients with diseased aorta and further studies should be pursued concerning Esmolol as an alternative to the conventional cardioplegia. PMID- 9341266 TI - [Changes in neutrophil counts and lymphocytes subpopulations of a ECLS instituted patient]. AB - A 4-year-old girl with tetoralogy of Fallot developed acute heart failure after ASD semiclosure. As drugs had no effect, ECLS was instituted. She gradually recovered from acute heart failure. ECLS was detached at 5 days after institution. Neutrocytopenia and lymphocytopenia became apparent during ECLS institution. The Subpopulations of T cell and NK cell decreased, and B cell subpopulation increased on the contrary during ECLS institution. This lymphocytopenia was caused by a decrease in T cell, especially CD4(+) cell numbers. It is necessary to minimize the potential for infection during ELCS institution. PMID- 9341267 TI - [DDD pacemaker implantation--cardiac pacemaker tips in the left atrium and ventricle by the left thoracotomy]. AB - Recently DDD pacemaker implantation for the children has undergone trials world wide; though regarding the approach, ways and positions of the epicardial lead, a few problems are still remained to be discussed. Now we report 9 cases (5 males, 4 females) of DDD pacemaker implantations by the left anterolateral thoracotomy approach. The 9 patients weighing 6.5 to 33 kg, were aged 11 months to 12 years (mean 6 years) of whom male 5, female 4 with degree of Block; 2 and 7. To all patients the stab-in type epicardial tips were implanted in the left atrium, the screw-in type ventricular epicardial tips were in the left ventricle by the 4th intercostal thoracotomy, and the pacemaker generators were beneath the fascia of the abdominal rectus muscle. We have no sensing and pacing failure, all pacemakers are working in the DDD mode well. PMID- 9341268 TI - [Thoracoscopic enucleation of esophageal leiomyoma]. AB - We treated four cases of thoracoscopic enucleation of esophageal leiomyoma. All four cases were asymptomatic, but either barium swallow or esophagofiberscopic examination revealed esophageal submucosal tumor. The locations of the tumors were middle and lower in one case and middle in the other three cases. All patients were intubated with a double lumen endotracheal tube under general anesthesia. Two patients required thoracotomy due to the tumor surrounding the esophageal wall in one case and severe adhesion to the esophageal mucosa in the other. The mini-thoracotomy was used in three cases. In the other two cases, we used four and three trocars, respectively. The balloon catheter, which had been inserted into the esophageal lumen, was useful for removing the tumor. The tumor was pulled up using the traction suture and dissected from the mucosa and muscular layer. After enucleation of the leiomyoma, the split muscular layer was sutured. The postoperative course was uneventful. These two patients were discharged on the 12th and 15th postoperative days, respectively. We conclude that the thoracoscopic enucleation of the esophageal leiomyoma is useful for reduction of surgical stress and is a more feasible approach for the treatment of esophageal leiomyoma. PMID- 9341270 TI - [A surgical case of intrapericardial inferior vena cava injury complicating liver injury caused by a blunt trauma]. AB - A 63-year-old man was brought to our hospital 20 minutes after sustaining blunt injury. He had been struck by a steel frame weighing about 700 kg that had fallen from a height of 2 meters. On admission, he was in cardiogenic shock with a systolic pressure of 70 mmHg and a pulse rate of 130 beats/minute. There was no apparent open wound and no brain injury was suspected. Echocardiography and chest computed tomogram showed cardiac tamponade, and abdominal computed tomogram showed liver injury. We performed emergency operation under cardio-pulmonary bypass standby. Laceration of the inferior vena cava at the right atrium junction was noticed. We had to perform IVC repair under cardio-pulmonary bypass because of massive bleeding. Liver injury was repaired after neutralization of systemic heparinazation. The patient developed respiratory, hepatic and renal failure during his postoperative course; however, he was discharged on postoperative day 39. Intrapericardial IVC injury caused by blunt trauma is a very rare event. Reconstruction can be performed successfully under cardio-pulmonary bypass. PMID- 9341269 TI - [Tricuspid valve replacement for infectious endocarditis associated with ventricular septal defect--report of three cases]. AB - We reported two male and one female patient (17, 36 and 47 years old, respectively) who presented infectious endocarditis (IE) in association with ventricular septal defect (VSD). In all cases, surgical treatment was performed in the acute stage of IE for persistent sepsis, pulmonary embolisms, and for giant vegetations. Because the tricuspid valve apparatus was severely damaged, valve replacement with the Carpentier-Edwards pericardial bioprosthesis was done and small VSD was directly closed in all cases. In one case, a complete heart block occurred, which necessitated postoperative implantation of a permanent pacemaker. All patients recovered and resumed their original social activities without the relapse of endocarditis. PMID- 9341271 TI - [Primary pulmonary malignant lymphoma of mucosa-associated lymphoid tissue (MALT) -a case report with a review of Japanese literatures]. AB - A case of non-Hodgkin malignant lymphoma of the lung was presented with a review of Japanese literatures. A 53-year-old woman was referred to our hospital because of an abnormal shadow on the roentgenogram of mass screening, with neither subjective symptoms nor abnormalities in physical examinations. Laboratory tests showed normal values. The diagnosis of pulmonary malignant lymphoma was obtained by immunohistochemical examinations. She underwent a upper lobectomy of the left lung with lymph nodes dissection of the mediastinum. The histological immunohistochemical and diagnosis of the resected tumor was primary pulmonary B cell lymphoma of mucosa-associated lymphoid tissue (MALT) without regional lymph nodes involvement. She has been doing well without any signs of recurrence for 15 months after the operation. PMID- 9341272 TI - [A case of traumatic cardiac contusion accompanied by the rupture of pericardium]. AB - We experienced a case of traumatic cardiac contusion accompanied by the rupture of the pericardium after multiple blunt trauma sustained in a traffic accident. A 26-year-old woman who had suffered from blunt chest and abdominal trauma was admitted to our hospital, being unconscious with multiple severe injuries including pelvic fracture, bilateral hemothorax, and multiple fractures in the extremities. The patient was in a shock status. We performed the transcatheter arterial embolization of the internal iliac arteries to control the bleeding, when aortography showed that the contrast media extravasated toward the left thoracic cavity. Immediately, an operation for blunt chest trauma was performed. Blood was flooding out of the ruptured pericardium because of the contusion of myocardium. The postoperative course was uneventful. Blunt chest trauma is usually accompanied by multisystem injury. Therefore, it is imperative to determine the priority of treatment based on preoperative examination in patients having multiple injuries. PMID- 9341273 TI - [A case report of left postero-lateral thoracotomy for simultaneous CABG and left lower lobectomy]. AB - Surgical management of patients with concomitant resectable lung lesions and critical cardiac disease is controversial. We report a case of concomitant pulmonary and cardiac surgery via a left thoracotomy. A 67-year-old male was admitted to our hospital complaining of recurrent bloody sputum and an abnormal shadow on chest X-ray. Chest CT and MRI showed a tumor in the left lower lobe (S10), with invasion of the diaphragm. A diagnosis of squamous cell carcinoma was obtained by transbronchial lung biopsy. The patient had a history of angina pectoris, and stress testing was positive. Coronary angiography showed 90% stenosis at segment 5, suggesting a risk of perioperative or postoperative myocardial infarction. This necessitated simultaneous surgical treatment for lung cancer and ischemic heart disease. A lobectomy of the left lower lung was performed, followed by coronary artery bypass grafting (CABG), using the great saphenous vein. The postoperative course was uneventful except for the occurrence of cholecystitis. Lung cancer and ischemic heart disease can be safely treated simultaneously via a single incision, with and benefit for selected patients. PMID- 9341274 TI - [Two resected cases of cavitary lung cancer with pulmonary aspergillosis]. AB - Two cases of cavitary lung cancer with pulmonary aspergillosis were experienced. Case 1 was a 45-year-old male. Chest X-ray and Chest CT revealed a round shadow in the thin-wall cavity of the upper lobe of the right lung. Upper lobectomy of the right lung was performed. Histologically large cell carcinoma was found to invade the entire cavity wall, and aspergillus was not detected in the intracavitary space. Case 2 was a 75-year-old male. Chest X-ray and Chest CT revealed a round shadow in the thin-wall cavity of the upper lobe S1 + 2 of the left lung. As a result of upper lobectomy of the left lung and S6 partial resection, large cell carcinoma was found to invade the entire cavity wall, and aspergillus was not detected in the intracavitary space. Only 19 cases including ours are reported about cases of lung cancer complicated by pulmonary aspergillosis at the same site in Japan. The mechanism of aspergillus infection had not been clarified in the discussions of the reported literature and nothing characteristic could be pointed out in our cases except for the assumption that the presence of cancer was a factor triggering Aspergillus implantation. PMID- 9341275 TI - [One-stage replacement of the entire thoracic aorta with aortic valve reimplantation technique--a reoperation for a Marfan patient with annulo-aortic ectasia and chronic aortic dissection of DeBakey type I]. AB - A 23-year-old man with Marfan syndrome, who had annulo-aortic ectasia and chronic aortic dissection of type I, was successfully treated. He underwent one-stage replacement of the entire thoracic aorta using a retrograde pull-through technique with aortic valve sparing reimplantation (David procedure). The descending aorta was replaced with a Hemashield graft, and then the graft was passed through within the descending aortic aneurysm in the retrograde fashion. Thus, the graft was inserted inside the descending aortic aneurysm without ligation of some of the intercostal arteries. Postoperative MRI showed complete clotting of the space between the graft and the aneurysm. David's reimplantation procedure which spares own aortic valve, requires no anticoagulant therapy in the post operative period. Neither aortic regurgitation nor pressure gradient between the left ventricle and the aorta were observed postoperatively. We conclude that the replacement of the entire thoracic aorta using a retrograde pull-through technique with an aortic valve sparing reimplantation by the single stage is useful for the selected patients with aneurysm of the entire thoracic aorta and annulo-aortic ectasia. PMID- 9341276 TI - [One stage operation for sternal turnover with preserved rectus muscle pedicles and aortic root replacement associated with Marfan's syndrome]. AB - A 40-year-old man with Marfan's syndrome had annulo aortic ectasia with Sellers grade 4 aortic valve regurgitation and Wada grade 3 pectus excavatum. Simultaneous operation was successfully performed by aortic valve composite graft insertion and sternal turnover with the rectus muscle pedicles. Following a midline skin incision, the cost-sterno complex (plastron) was dissected together with the bilateral rectus muscle pedicles, and the sternum was divided transversely through the second intercostal space. The plastron with muscle pedicles was retracted away from the anterior chest toward the abdomen and was covered by the moistened sternal bag made of polyethylene to prevent dryness and contamination during the composite graft insertion. The aortic root was replaced with a composite graft consisting of a 25 mm SJM valve and a 26 mm Hemashield graft. A short interposed 10 mm Hemashield graft was inserted between the ostia of the left coronary artery and the composite graft. The right coronary artery was reimplanted in the aortic conduit using the button technique with a doughnut pledget. This one stage method offered excellent operative exposure and enabled us to prevent possible necrosis of the sternum, infection of the mediastinal sinus, and postoperative cardiac failure resulting from chest wall compression. In this procedure, active usage of the rapid autologous transfusion system effectively reduced the total amount of blood transfusion. PMID- 9341277 TI - [Ruptured thymic cysts with mediastinal hemorrhage and hemothorax--a case report and reviews of the literature]. AB - We report a rare case with rupture of thymic cyst including mediastinal hemorrhage and hemothorax. A 68-year-old man was referred to our hospital for the treatment of right hemothorax. A chest roentgenogram following thoracentesis demonstrated a widened mediastinum. Chest computed tomograms revealed a large anterior mediastinal mass extending to the right pleural cavity and bilateral pleural effusion. Digital subtraction angiography showed a normal aorta and great vessels. A median stenotomy revealed a large encysted hematoma along the thymic cyst extending from the right anterior mediastinum into the right pleural cavity and ending with rupture. Pathological examination demonstrated that the largest thymic cyst was continued to the hematoma resulting from partial destruction of its epithelial lining. To our knowledge, only five cases including our's with ruptured thymic tumors were reported, and this is the first report of ruptured thymic cyst resulting in mediastinal hemorrhage and hemothorax. PMID- 9341278 TI - [Associations between breathing pattern during submaximal exercise and exercise capacity in patients with pulmonary emphysema]. AB - We sought to clarify the factors associated with exercise capacity in patients with pulmonary emphysema. Exercise capacities of 20 men with pulmonary emphysema were evaluated by bicycle ergometery, and the results were used to divide the subjects into two groups: high exercise capacity (n = 10) and low exercise capacity (n = 10). Pulmonary-function tests were done, emphysema scores were computed from CT scans, breathing pattern was recorded during submaximal exercise (up to 20 watts), and index of rapid shallow breathing was computed. Neither FEV1 nor airway resistance differed between the two groups, and patients with lower exercise capacity tended to have lower tidal volumes and higher values of the index of rapid shallow breathing during submaximal exercise. Functional residual capacity measured by body plethysmography and emphysema scores were inversely associated with exercise capacity. We speculate that among patients with pulmonary emphysema and a given degree of airway obstruction, a high functional residual capacity causes breathing during submaxinal exercise to be rapid and shallow, and that this rapid and shallow breathing makes ventilation inefficient, increases the work of breathing, and limits exercise capacity. PMID- 9341279 TI - [Prediction of outcome after acute exacerbation of idiopathic interstitial pneumonia]. AB - Some patients with chronic idiopathic interstitial pneumonia (IIP) experience acute exacerbations (AE). Because the precise mechanisms of AE in patients with IIP remain unclear, the treatment for AE is not established and the efficacy of steroids is controversial. Consequently, it is difficult to predict outcomes in patients with AE of IIP. We therefore studied the relationship between clinical findings, efficacy of treatment, and clinical outcome in patients with AE of IIP. Thirty-two patients were enrolled, and were divided into two groups: 10 who were alive more than one year after the onset of the AE survivors, 8 men and 2 women, and 22 who died within one year of the AE (non-survivors, 17 men and 5 women). Survivors were significantly younger than non-survivors (59.7 +/- 9.9 vs 67.5 +/- 8.2 years, respectively, p < 0.05). The values of PaCO2 measured before the AE were higher in survivors than in non-survivors (43.0 +/- 3.7 vs 38.4 +/- 4.0 torr, respectively, p < 0.05). At the onset of the AE the levels of C-reactive protein in serum were higher in survivors than in non-survivors (13.9 +/- 7.9 vs 7.3 +/- 5.8 mg/dl, respectively, p < 0.05). Chest X-ray films showed progression of ground-glass shadows in both groups when the AE occurred; the radiographic findings did not differ markedly between groups. Of the 22 non-survivors, 7 had received medication before the AE; none of the survivors had received medication before the AE. At the time of the AE all patients were treated with steroid pulse therapy, and the dose of methylprednisolone used, did not differ significantly between groups. These data suggest that three factors are closely related to responsiveness to steroid therapy and to clinical outcomes after AE in patients with IIP: 1) age at the onset on IIP, 2) respiratory status before the AE, and 3) disease activity as reflected by inflammatory reactions. PMID- 9341280 TI - [Six patients in whom exacerbation of asthma was complicated by mediastinal emphysema]. AB - We encountered 6 patients with bronchial asthma complicated by mediastinal emphysema. Their average age was 21 years. All were atopic and had histories of asthma in childhood. In 5 of the 6, mediastinal emphysema developed when the patients had upper respiratory infections. In 3, the mediastinal emphysema worsened. After insertion of subcutaneous drains to decrease intramediastinal pressure, the conditions of those 3 patients improved remarkably. The conditions of the other 3 improved after medication only. During the period of this study, 1.24% of all patients treated at Hiroshima Asa Citizen's Hospital for exacerbations of asthma had mediastinal emphysema. PMID- 9341281 TI - [Transfer of genes to human airway epithelial cells in vitro via DNA virus vectors]. AB - Transfer of genes via DNA virus vectors to human airway epithelial cells is not fully understood. We tested the effects of multiplicities of infection (moi) and incubation time on the efficiency of gene transfer by adenovirus vectors and AAV vectors in cultures of human airway epithelial cells. The cells were exposed to Ad-CMV-lacZ or AAV-CMV-LacZ for 1 to 48 hours at different moi from 0.1 to 100. Efficiency of transduction was assessed as the percentage of cells that expressed LacZ, and was measured by using X-gal staining. A dose-dependent relationship was found between vector moi and the percentage of cells that expressed LacZ. For both vectors, gene expression was greater when incubation time was longer (up to 12 hours), which suggests that the uptake of vectors into the cells is not selective. Efficiency of transduction did not differ between adenovirus and AAV vectors at the same moi. These results suggest that (1) gene transfer via DNA virus vectors increases in a moi-dependent fashion, and that (2) gene expression of the DNA virus vector may be increased by prolonged exposure of the vector to human airway epithelial cells. PMID- 9341282 TI - [Steroid resistance and lung-tissue cytokines in experimental bleomycin-induced lung fibrosis]. AB - To examine the mechanism of steroid resistance in lung fibrosis, cytokines expressed in the lung tissue of mice with bleomycin-induced lung fibrosis were studied. 1. Glucocorticoid administration (1 mg/kg/day) did not affect the grade of lung fibrosis induced by intratracheal injection of bleomycin (3.76 micrograms/g). 2. Cytokines expressed in the lung tissue were studied with the reverse-transcriptase polymerase chain reaction. Levels of promotor cytokines, such as TNF alpha, TGF beta, INF gamma, and IL-2, were significantly higher in lung tissue from the bleomycin group. The expression of these cytokines in the glucocorticoid group was low, especially the peak value. Expression of IL-4 was high in the bleomycin group, and was not inhibited in the glucocorticoid group. Expression of the down-regulator cytokine IL-10 was also high in the bleomycin group and very low in the glucocorticoid group. 3. The non-selectivity of glucocorticoids with respect to promotor and suppressive cytokines may account in part for steroid resistance in bleomycin-induced pulmonary fibrosis. PMID- 9341284 TI - [Allergic bronchopulmonary aspergillosis in a patient with no symptoms of asthma until after bronchial lavage]. AB - An asymptomatic 56-year-old man was admitted to our hospital because of an abnormal shadow on a chest X-ray film. Allergic bronchopulmonary aspergillosis was diagnosed on the basis of five findings: eosinophilia, immediate skin reactivity to Aspergillus antigen, the presence of precipitating antibodies against Aspergillus antigen, a high concentration of IgE in serum, and central bronchiectasis. He had no symptoms of asthma at the time of diagnosis, but did a few days after he underwent bronchial lavage. We speculate that the asthma attack was related to the bronchial Lavage as follows: First, drainage of mucus plugs by bronchial lavage may have exposed the bronchial epithelium, which had already been sensitized, to aspergillus antigens. Second, the scattered antigen may have dose-dependently stimulated the bronchi. Third, the infection may have increased bronchial responsiveness to the antigen. Symptoms of bronchial asthma are not necessary for the diagnosis of allergic bronchopulmonary aspergillosis. PMID- 9341283 TI - [Plasma cell interstitial pneumonia as a manifestation of multicentric Castleman's disease]. AB - A 12-year-old boy was admitted to our hospital because of abnormal shadows on a chest radiograph, slight fever, and superficial lymphadenopathy. Laboratory examination showed anemia (Hb 9.9 g/dl) and hyperimmunoglobulinemia (IgG 5469 mg/dl) without M protein. A chest CT scan showed bilateral diffuse shadows and bilateral hilar lymphadenopathy. Biopsy specimens of an inguinal lymph node and a lung showed many lymphoid follicles with germinal centers, and marked infiltration of mature plasma cells in the interfollicular area without destruction of follicular structures. The polyclonality of the plasma cells was confirmed by immunohistochemistry. The patient was not treated because these results excluded malignant disease and he was asymptomatic. At the age of 17 years, he was admitted to our hospital again because of dyspnea and a tendency to bleed. Interstitial pneumonia, hyperimmunoglobulinemia (IgG 13900 mg/dl), and anemia (Hb 6.6 g/dl) were found, along with thrombocytopenia (2.5 x 10(4)/mm3) and proteinuria. The serum interleukin-6 level was high: 177 pg/ml. Bronchoalveolar lavage fluid contained many plasma cells. Therapy with corticosteroids and immunosuppressant medication was effective. Our diagnosis was plasma cell interstitial pneumonia as a manifestation of multicentric Castleman's disease. PMID- 9341286 TI - [Mediastinal esophago-bronchogenic cyst presenting as a single mass]. AB - An abnormal shadow was found on a chest X-ray film of a 42-year-old man. The mass was 17 x 12 x 11 cm, smooth, round, homogeneous, and was seen in the left upper lung field. Chest computed tomography revealed a cystic mass in contact with the left lung, the chest wall, the esophagus, and the aorta. Magnetic resonance imaging showed a wall that divided the mass into two compartments. The cystic lesion was removed and was found to contain a yellow liquid. The larger part of the mass was histologically similar to esophageal wall and the smaller part was bronchogenic. The final diagnosis was mediastinal esophago-bronchogenic cyst. In previously reported cases, two cystic masses were connected by a canal. The findings in this case support the theory by Yoshii that this type of cyst originates from the diverticulum of the foregut near the lung bud. PMID- 9341287 TI - [Infected emphysematous bullae in a patient with diabetes mellitus]. AB - A 42-year-old woman with diabetes mellitus was admitted to our hospital because of fever, coughing, and dyspnea. Coarse crackles were audible and respiratory sounds were weak in the right lung field. Laboratory examination revealed a high erythrocyte sedimentation rate, a high level of serum C-reactive protein, a high blood sugar level, and hypoxemia. A chest roentgenogram revealed cystic lesions with fluid levels, and an infiltration shadow in the right lung field. A chest computed tomographic scan revealed many cystic lesions with fluid levels and an infiltration shadow. Our diagnosis was infected emphysematous bullae. A tube was inserted percutaneously for drainage and to allow injection of antibiotics into the cystic lesion. The cystic lesion then vanished. Percutaneous drainage and washing with antibiotics can be used to treat infected emphysematous bulla that have thick closed cystic walls. PMID- 9341288 TI - [Primary pulmonary non-Hodgkin's lymphoma in a patient with dermatomyositis]. AB - A 47-year-old man with dermatomyositis and interstitial pneumonia had been treated with prednisolone since May, 1992, and with azathioprine since April, 1993. During the sixth month of this treatment, primary pulmonary non-Hodgkin's lymphoma (T-cell, diffuse, pleomorphic) developed. Chemotherapy (vincristine and adriamycin) was begun but there was no response. An invasive lesion of the brain was seen on a CT image. Despite cranial radiotherapy, the patient died of respiratory suppression due to progressive brain disease on December 14, 1993. Primary pulmonary non-Hodgkin's lymphoma develops only rarely in patients with dermatomyositis. In this case, oncogenesis may have been related to the use of immunosuppressants. PMID- 9341285 TI - [Congenital antithrombin III deficiency associated with pulmonary thromboembolism]. AB - A 42-year-old man was admitted to the hospital because of sudden left-lateral chest pain and dyspnea. The chest roentgenogram on admission showed a string-like shadow and pleural effusion, and analysis of arterial blood gases revealed hypoxemia. Many bilateral segmental defects were seen on a radioisotope perfusion scan of the lungs, and the chest CT scan showed a low-density area in the trunk of the pulmonary artery. From these findings, pulmonary thromboembolism was diagnosed. The serum antithrombin III (ATIII) activity and the antigen value were low, and the patient's son's serum ATIII level was also low. Both the patient and his son were given the diagnosis of congenital ATIII deficiency. Gene analysis revealed two point mutations in exon 6 of the ATIII gene, so we classified this case as type IIc. The patient was treated with intravenous urokinase and heparin, and was then given warfarin. The arterial blood gas tensions improved, and the low-density area of the pulmonary artery on the CT scan disappeared, as did the pulmonary perfusion defect on the radioisotope scan. The clinical manifestations of members of 28 families with congenital ATIII deficiency reported in Japan were reviewed. Patients in whom many point mutations were detected in the ATIII gene were rare. PMID- 9341289 TI - [Levels of serum KL-6 in a patient with drug-induced pneumonitis]. AB - We describe a 67-year-old man in who serum KL-6 levels were measured during drug induced pneumonia. The patient was hospitalized, because of coughing, fever, and dyspnea on exertion after administration of Sho-saiko-to (herbal medicine). After he was hospitalized, his symptoms were relieved, and the infiltration shadow on chest X-ray films resolved, but after re-administration of Sho-saiko-to, fever and hypoxemia developed. The serum KL-6 level was again high one day after oral re-administration of the drug. However, the level of lactate dehydrogenase in serum was not high after the re-administration. After treatment with on oral steroid drug the serum KL-6 level decreased gradually, symptoms were relieved the previously high level of c-reactive protein in serum decreased, the previously high white blood cell count decreased, and radiographic findings returned to normal. The diagnosis of drug-induced pneumonia is difficult, because specific diagnostic measures have not been developed. In the present case the serum KL-6 level increased rapidly after re-administration of the drug, and therefore measurement of serum KL-6 level may be helpful in the diagnosis of drug-induced pneumonia. PMID- 9341290 TI - [Interstitial pneumonia caused by manidipine HCl]. AB - A 62-year-old woman was given a diagnosis of rheumatoid lung in 1993. She began receiving manidipine HCl (10 mg per day) on June 19, 1996 to treat hypertension. The next day fever, coughing and dyspnea developed. She was admitted to our hospital on June 28. A chest radiograph showed diffuse reticulo-nodular shadows in all lung fields and arterial blood gas analysis revealed severe hypoxemia. Administration of manidipine HCl was stopped and treatment with methylprednisolone was started. The symptoms and the radiographic evidence of infiltrates disappeared. A drug lymphocyte stimulation test for manidipine HCl was positive. We know of no previous report of pneumonia caused by manidipine HCl. PMID- 9341291 TI - [Cardiac tamponade caused by diffuse pericardial mesothelioma]. AB - A 75-year-old woman was admitted to our hospital because of dyspnea and fever. A chest roentgenogram obtained on admission showed cardiomegaly. An echocardiogram, a computed tomogram, and a magnetic resonance computed tomogram revealed a pericardial tumor and a large pericardial effusion. A tumor biopsy was done under echocardiographic guidance, and sarcomatous mesothelioma was diagnosed on the basis of histological and immunohistological studies of biopsy specimens. Pericardial fenestration followed by tumor resection gave relief from cardiac tamponade. The postoperative course was good, and the patient was discharged from the hospital on the 19th postoperative day. Four months later the pericardial mesothelioma recurred and the patient died of constrictive pericarditis. Palliative resection was useful in this case because it allowed the patient to resume activities of daily living by relieving the cardiac tamponade. PMID- 9341292 TI - Quantitative morphology of dermal elastic fibers system of the human face during aging. AB - Human skin has various distributions and arrangements of elastic fiber (EF). Previous reports did not clearly show the distribution of EF in the face skin because of various contents during aging. In this study, a color image analyzer indicated distribution of elastic, oxytalan, and muscle fibers in human face skin. During aging the muscle fiber size and the content of the EF decreased in the modiolus and inferior labial regions of the human skin, and the ratio of the EF was lower than that of oxytalan fiber measured areas. That is, the dimension of oxytalan fiber may reflect the content of EF, and muscle has a role in the distribution of the EF in human face skin. In the deeper regions, small and large EF bundles were found near the sheath of gland and muscles. Therefore, face movement might be an important aspect to maintaining the EF content of human face skin. PMID- 9341293 TI - Electron microscopy of the senile changes in lens epithelium. AB - Lens epithelia obtained from 37 patients who had undergone cataract surgery operation were examined by using transmission electron microscope. Their ages varied between 60-79 years and mean age was 70.2 years. Additionally, the lens epithelia of 14 patients, which were obtained in the same way were examined by using scanning electron microscope. Their ages varied between 58-78 years and mean age 70.7 years. In transmission electron microscopy, the normal appearing epithelial cells were intermingled with abnormals and the abnormal cells increased in number and in degree of abnormality with aging. The three dimensional structure of the interdigitating interlocking processes of lens cuboidal cells were visualised by using scanning electron microscope. PMID- 9341294 TI - The effect of magnesium on the fine structure of the golden hamster parathyroid gland in vivo and in vitro. AB - We examined the effect of magnesium (Mg) on the fine structure of the golden hamster parathyroid gland in vivo and in vitro. In the in vivo study, the principal changes in the parathyroid glands 10 and 30 min, and 1 hr after Mg injection showed a significant decrease in the area of the Golgi apparatus and cisternae of the rough endoplasmic reticulum compared with those of the control animals. The serum calcium level in the experimental animals 30 min and 1 hr after the injection was significantly low compared with that of the control animals. In the vitro study, the area of the Golgi apparatus in the Mg-treated group significantly decreased as compared with that of the control group. These changes suggest that Mg directly suppresses the synthesis of hamster parathyroid hormone in the short term. PMID- 9341295 TI - A case simultaneously presenting with a rare portal collateral pathway and left gastric venous anomaly. AB - We had the opportunity to dissect an autopsy case who had developed a rare portal collateral pathway due to increased portal pressure resulting from liver cirrhosis and simultaneous abnormal left gastric venous distribution. The portal collateral pathway consisted of a well-developed communicating branch located between the left renal vein and the left gastric vein. The left gastric vein did not merge into the portal vein, but directly entered the liver after bifurcating near the hepatic hilum. One branch had an anastomosis to the left branch of portal vein in the liver and the other distributed in the hepatic quadrate lobe. We considered this aberrant left gastric vein to be a congenital residue of the embryological left portal vein. The present case is the third Japanese case to have been described minutely in the literature, following the two cases reported by Miyaki et al. (1987). Persistence of the umbilical vein and the absence of the celiac trunk were also observed. PMID- 9341296 TI - Fluoride-resistant acid phosphatase (FRAP)-positive afferent terminals make synaptic contact with interneuronal soma in the substantia gelatinosa of the mouse spinal dorsal horn. AB - Fluoride-resistant acid phosphatase (FRAP)-reactive terminals making contact with interneuronal soma are found in the substantia gelatinosa of the mouse spinal dorsal horn. About one half of the interneuronal somata receive FRAP-positive boutons. By electron microscopy, these FRAP-positive terminals appear small, dark, slender, roundish, cap-like, ellipsoid or sinuous and electron-dense, scalloped (fan-like) contours with clear spherical synaptic vesicles of variable size, some large dense-core vesicles and mitochondria. All these features are very similar to those of capsaicin-sensitive terminals. Thus they are considered to be nociceptive primary afferent endings. Therefore, some of the FRAP-positive terminals are suggested to have a modulatory role in the nociceptive circuit in the substantia gelatinosa. PMID- 9341297 TI - Strength in numbers. PMID- 9341298 TI - Stop the madness. PMID- 9341299 TI - The sun also sets. PMID- 9341300 TI - Do something ... anything. PMID- 9341301 TI - Surprise decision. PMID- 9341303 TI - Hands-on science. PMID- 9341304 TI - Standing sober. PMID- 9341302 TI - What matters to your patients. PMID- 9341305 TI - Hantavirus pulmonary syndrome in the Americas. PMID- 9341308 TI - Sensory integration in the learning of an aiming task. AB - Results of recent research on motor control indicate that in an aiming task, visual information remains of primary importance for optimal accuracy even after extended practice. One of the points that is yet unclear is whether it is solely the dynamic visual information about the moving limb that is important for movement control. To shed some light on this issue, subjects practiced an oscilloscope aiming task. In a transfer test, the dynamic information regarding the displacement of the to-be-moved object could be withdrawn without altering the static visual information that had been available during the learning of the task. The results indicated that, after 200 trials of practice, withdrawing dynamic visual information regarding the displacement of the to-be-moved object produced a deterioration in the accuracy of the subjects' responses. This indicates that the role played by the dynamic visual information for aiming control does not diminish with practice. Moreover, although visual cues available before or after movement execution have been shown to help better plan an upcoming movement, the static cues available during movement execution do not appear to play an important role in the movement representation thought to develop through practice. PMID- 9341307 TI - Comparison of two surveys of hospitalization: the National Hospital Discharge Survey and the NHANES I Epidemiologic Followup Study. AB - OBJECTIVES: This report compares hospitalization data from the NHANES I Epidemiologic Followup Study (NHEFS) with data from the National Hospital Discharge Survey (NHDS), the benchmark for hospitalization in the United States, for men and women 35 years and older for the period 1971-87. The comparison is intended to help analysts evaluate the validity and generality of analyses based on the NHEFS. METHODS: Hospital stays per 1,000 population and average lengths of stay are compared year by year for each age-sex group and for the entire period. Regression analyses test for differences between the two surveys by age and sex, and for differences in trends over time and the effect of the Medicare program's prospective hospital payment system. RESULTS: Hospital stays per 1,000 population were lower in NHEFS than in NHDS in all age-sex groups at the beginning of the period, but the differences had almost disappeared by 1987. Lengths of stay, although somewhat longer in NHEFS, matched NHDS more closely. Differentials by age and sex were similar in the two surveys for both hospital stays per 1,000 population and length of hospital stay. With its extensive information on baseline risk factors, the NHEFS offers a unique opportunity to study determinants of hospitalization in a representative sample of U.S. adults. The evaluation presented here suggests two points for researchers who want to use the NHEFS. First, including age as a control should largely correct for differences in age distribution between NHEFS and NHDS. Second, a time trend should also be included to capture the effects of several factors that caused the count of stays to be low in the early years of NHEFS followup. PMID- 9341309 TI - Over the counter medications in the workplace and legal liability. PMID- 9341310 TI - AAOHN position statement. Natural rubber latex sensitivity. PMID- 9341311 TI - Risk of tuberculosis transmission in dentistry. Results of a retrospective chart review. AB - This pilot project was conducted for the purpose of performing a retrospective chart review on selected clients and using the results for evaluating purified protein derivative (PPD) conversion rates among the student population. The occupational risk of exposure to active tuberculosis was assessed in a large dental educational setting. Charts of clients seen in the College's Oral Medicine Clinic, referred out for health care consultation for one of several reasons potentially associated with active tuberculosis disease over a 1 year period, were reviewed. Data sources included the medical consultation log and the tuberculosis log, which were maintained by faculty in the Oral Medicine Clinic. Ninety-six clients met the authors' criteria. However, compiling data was severely hampered for two reasons: missing charts (19 of 96, or 19.8%) and non returning clients (55, or 57%). Four clients with potentially active cases of tuberculosis were identified. Follow up revealed, however, that none of these four clients was contagious when seen at the Dental Center. The protocol and definition recommended by the Centers for Disease Control and Prevention 1994 Guidelines, and the results of PPD screening and chart audit conducted by the authors, suggest that the employees and students of the College of Dentistry are at low risk for workplace exposure to active tuberculosis. PMID- 9341312 TI - Animal exposure risks. Implications for occupational health nurses. AB - 1. Diseases transmitted from animals to humans, referred to as zoonoses, pose risks for persons occupationally exposed to animals. 2. Occupational health nurses should be proactive in identifying potential occupational health risks and educating employees to protect themselves from animal exposure risks. 3. Exposure history should include exposure to birds and animals during occupational and recreational activities. 4. Primary prevention is critical, as immunizations and treatments are limited and include many associated risks. PMID- 9341314 TI - Noise exposures. Effects on hearing and prevention of noise induced hearing loss. AB - 1. Over 30 million workers are exposed to hazardous noise on the worksite. Continual exposure to high noise levels damages and destroys hearing cells within the ear, making noise induced hearing loss an irreversible impairment. 2. Hearing conservation programs are required by law for workers in industrial settings where noise exposures equal or exceed 85 dB(A). Many workers, such as those in construction and agricultural industries, are not covered by these programs. 3. Reducing noise through engineering or administrative controls is the first line of defense. When this is not sufficient, two types of personal hearing protection devices are available: passive hearing protection devices such as ear muffs, canal caps, and ear plugs, which reduce noise mechanically; and active noise reduction devices, which electronically cancel sound waves at the ear. 4. The most effective hearing protection devices are those with which the worker is most comfortable will use 100% of the time. The occupational health nurse has a major role in promoting increased use of hearing protection devices through continued contact with workers, administrators, and safety personnel. PMID- 9341313 TI - Farm worker injuries associated with bulls. New York State 1991-1996. AB - 1. Although cows greatly outnumber bulls on dairy farms, bulls account for 25% of animal related injuries in a surveillance study of agricultural injury. In addition, bull injuries are more severe. 2. Because of their size and unpredictable behavior, bulls, especially those over 18 months of age, must be handled with extreme caution. 3. Important risk factors for the observed incidents were working alone and not having an escape route. 4. Bulls should be dehorned and confined in specially designed facilities to avoid human contact during feeding, watering, exercising, or breeding. PMID- 9341315 TI - Linda Rosenstock, NIOSH director, talks about NIOSH's relationship with occupational health nursing. Interview by Eileen Lukes. PMID- 9341316 TI - Business skills for the occupational health nurse manager: conducting an effective interview. PMID- 9341324 TI - The saga of a journey ... and interludes. PMID- 9341325 TI - The North Carolina Association of Colored Graduate Nurses: a proud heritage. PMID- 9341326 TI - Holistic retention: a practical approach. AB - In this article the author discusses a retention program for students with multiple needs at Coppin State College, a historically Black institution. The author makes the point that students with multiple needs require a holistic approach if they are to be successful in schools of nursing. This approach is based on a focus of the total student being supported in a culturally sensitive, supportive environment for learning. PMID- 9341327 TI - The mystery of nursing. PMID- 9341328 TI - African American women and osteoporosis. AB - Osteoporosis is a condition that presents differing predisposing factors between European American Women and African American Women. Preventative health is commonly focused on European women because they are at higher risk. This article presents issues and facts that substantiate the need to also target African American women for osteoporosis prevention education. PMID- 9341329 TI - Increasing retention rates of disadvantaged students through a faculty development program. AB - Efforts to recruit disadvantaged students have been increasing over the past three decades. How to retain those students once they have been recruited remains a critical issue for faculty. The authors include a Faculty Development Model presently being implemented at an inner city community college to increase faculty's expertise in enhancing the skills and abilities of students from diverse backgrounds with an emphasis on the disadvantaged student. PMID- 9341330 TI - Mentoring for publication: faculty and student perspectives. AB - Mentoring can be used to teach the writing process and to encourage undergraduate minority nursing students to produce manuscripts for publication. Through a one to-one mentoring relationship with a nursing faculty member, a student is made aware of the links between writing as a critical skill for learning and acquiring the skills for professional nursing. The mentoring relationship supports the student from developing an idea for publication to actively participating in preparing a manuscript. PMID- 9341331 TI - The publication process: steps to success. AB - In this article, the author discusses the six steps anyone can use to have a successful writing career. She outlines each step and encourages her audience to participate at each level in order to be successful. The steps are: need/desire; expertise; write; peer review; submit; acceptance and/or revise and re-submit. PMID- 9341332 TI - History as communication. PMID- 9341333 TI - The founding of the Association of Black Nursing Faculty: my memories of the first five years. AB - This article reviews the organization and development of the Association of Black Nursing Faculty. Incorporated (ABNF). The author, who is also the founder, discusses her memories of the founding of the first professional organization for Black nursing faculty in the United States. PMID- 9341334 TI - Chattanooga and Cape Town: a cross continental medical/nursing partnership. AB - The author discusses her recent four-week visit to Cape Town. South Africa as a part of a multi-disciplinary, multi-racial, medical/evangelistic team to provide health related services to people in nine different sites. She also discusses her impressions of the country and the people from an African American perspective. PMID- 9341335 TI - Managing clinical experiences for minority students with physical disabilities and impairments. AB - According to the Americans with Disabilities Act of 1990, clinical training programs cannot refuse admission to individuals with certain physical limitations. That is, an otherwise qualified person who meets program requirements in spite of a handicap must not be discriminated against. Since hypertension and renal disease affect a disproportionately higher number of African Americans than others, the numbers of students with physical disabilities as well as students of color who are also adult learners are likely to increase in clinical nursing programs. This article discusses a planning model developed and implemented by nursing faculty in a baccalaureate degree nursing program at a historically Black university which was used as a guide for the inclusion in clinical practice of students experiencing physical disabilities. A case study is presented to illustrate how the model was implemented with an immunosupressed student secondary to a renal transplantation. PMID- 9341336 TI - Factors contributing to late hospice admission and proposals for change. AB - INTRODUCTION: Hospice seeks to provide high-quality, holistic end-of-life care. Most insurances will reimburse hospice care for a period of six to seven months. However, the majority of patients are not referred to hospice until they are very close to death. The purpose of this study was to examine characteristics of an urban hospice program that may be associated with inordinately late admission to hospice care. METHODS: A retrospective chart review of 100 consecutive admissions was carried out. Patient data from the referral/intake forms were reviewed and length of stay of each patient was calculated. RESULTS: The average length of stay in this hospice was 34 days. Over half of the patients (51 percent) died within 14 days, and more than one-third (35 percent) died within seven days of admission. African-Americans made up nearly one-third of the patient group (32 percent). A considerable number of patients had a non-cancer diagnosis (39 percent). More than half of the patients were referred from acute care hospitals/specialists (58 percent). CONCLUSION: The study shows that most patients were referred to this urban hospice program when they were very close to death. Three characteristics may contribute to this underutilization of the hospice benefit: (1) the large representation of African-Americans, (2) the high enrollment of patients with non-cancer diseases, and (3) the high number of referrals from acute care hospitals/specialists. Initiatives to overcome barriers to hospice care for minority populations, to better prognosticate terminal illnesses, and to educate physicians and patients about palliative care are needed so that more patients can benefit more fully from hospice care at the end of life. PMID- 9341338 TI - Hospice support for families facing multiple deaths of children. AB - The hospice team can have significant impact in guiding families through the death of a second child by recognizing the additional dynamics inherent in such a grieving process. It is vital to recognize the unresolved grief these families may face when they have not been able to grieve the first child's death adequately. The impact this unresolved grief will have on their coping abilities, anticipatory grief, and grief work when they experience the death of another child can be overwhelming. It is important to be aware of familial changes that occur when working with families who are facing the death of a second child since the addition of other children. For example, a host of additional dynamics that the family may not be familiar with or prepared to cope with may well occur, demanding specialized grief support and follow-up as provided by Hospice. PMID- 9341337 TI - Spiritual diversity: a challenge for hospice chaplains. AB - Spirituality is a critical component of the holistic mind-body-spirit model embraced by Hospice. Hospice chaplains, as part of the caregiving team, must understand their abilities and limitations in providing spiritual guidance to others who may differ in religious and spiritual beliefs. PMID- 9341339 TI - Pediatric hospice reference library. PMID- 9341340 TI - Reframing life's puzzle: support for bereaved children. PMID- 9341341 TI - Now I lay me down to sleep. Remembering Walter: a child preparing to die. PMID- 9341342 TI - Eulogy for Amanda. PMID- 9341343 TI - Hospice Northeast and Nemours Children's Clinic, Jacksonville, Florida. PMID- 9341344 TI - The Hospital for Sick Children, Toronto, Ontario, Canada. PMID- 9341345 TI - What is the Hospice Nurses Association? PMID- 9341346 TI - Asthenia in terminally ill cancer patients: a brief review. PMID- 9341347 TI - Dr. Dignity. PMID- 9341349 TI - The impact of patient anger and resentment on treatment decisions. PMID- 9341350 TI - Tailoring interventions for health behavior change in breast cancer screening. AB - PURPOSE: Health professionals continue to seek strategies to help individuals increase health-promoting behaviors and decrease those behaviors related to unhealthy lifestyles. Various health behavior theories currently guide interventions that focus on health behavior changes. Of these, a tailored approach uses individual data on beliefs and behavior to guide the content and delivery of interventions. The purpose of this paper is to explain the usefulness of theories in tailoring health promotion messages to increase health-protecting behaviors. OVERVIEW: A brief overview of current popular theories that focus on individual behavior is presented, followed by illustrations of a tailored intervention approach for breast cancer screening. Applications of the tailored intervention approach in ongoing studies to increase breast cancer screening are described. CLINICAL IMPLICATIONS: Clinical implications of the ongoing projects suggest practical and widespread utility of tailoring health-promoting messages to increase healthy lifestyles. The tailored approach may be used by a multidisciplinary team to focus on the individual's unique factors to impact behavior change rather than to deliver the same standard message to all individuals. PMID- 9341351 TI - Balancing demands of cancer surveillance among survivors of thyroid cancer. AB - PURPOSE: The purpose of this qualitative study was to explore and describe the demands of long-term cancer surveillance among survivors of thyroid cancer and how these perceived demands influence their quality of life. DESCRIPTION OF STUDY: The sample consisted of 34 participants who had undergone thyroid hormone withdrawal before body scanning for evaluation of cancer recurrence and/or metastatic disease. Participants completed two self-report instruments: 1) a Demographic Data Tool, and 2) three open-ended questions about quality of life during thyroid hormone withdrawal. The participants wrote down their answers to the demographics and open-ended questions. RESULTS: The 34 participants had a mean age of 40 years (range, 22-73 years); 85% were women and 74% were white. All were high school educated, and 32% were college graduates. Sixty-eight percent were married, and 75% were employed either full- or part-time. Participants experienced profound changes in relationship to withdrawal from thyroid hormone medication. These changes in physical well-being coupled with the potential for disease recurrence influenced their psychological and social well-being. Balancing the demands of cancer surveillance (through body scanning and thyroid hormone withdrawal) against day-to-day living had a major influence on perceptions of quality of life. CLINICAL IMPLICATIONS: Study results support other studies and clinical reports that physical symptoms related to thyroid hormone withdrawal were profound, severe, and debilitating. First, participants with thyroid cancer had to learn through their own personal experiences what physical limitations were imposed during the period of surveillance testing. Second, the physical changes and anticipation of body scanning exerted a profound effect on psychological and social well-being. Feelings of loss, anxiety, depression, and loss of concentration were very difficult to endure. Third, given the general characteristics of this population of thyroid cancer survivors who generally are younger and working, the experience of hypothyroidism can have a major impact on work schedules. Individuals who require a large amount of concentrated attention to work tasks may need to modify their work schedules during hypothyroid states. Findings suggest that individuals who are considered "cured" of their disease are a largely forgotten segment of cancer survivors. However, participants in this study continued to inform us of the process of cancer survivorship and the need for exploring quality-of-life concerns, regardless of prognosis. PMID- 9341352 TI - Recruitment of underserved women for breast cancer detection programs. AB - PURPOSE: The purpose of this correlational, nonexperimental study was to survey a sample of uninsured/underinsured women entering a no-cost, state-funded breast cancer screening program to identify factors that might enhance existing recruitment strategies and adherence to breast cancer screening. OVERVIEW: Inadequate access to breast cancer screening contributes to the high morbidity and mortality that have been documented in low-income American women with breast cancer. Two hundred and four participants in a no-cost cancer screening program at the Johns Hopkins Oncology Center's East Baltimore Community Health Program in Maryland were surveyed by telephone to identify methods to enhance recruitment strategies. The survey solicited information in areas such as demographics, healthcare practices, and recruitment sources. Knowledge about breast cancer and its treatment, perceived control over health matters, and the practice of preventive measures were low in this group. Based on the data obtained, recruitment strategies better tailored to this population were developed and implemented. CLINICAL IMPLICATIONS: Recruiters and clinicians must understand that for impoverished women, day-to-day survival takes precedence over most other issues. The possibility of a condition that may cause illness or death 10 to 20 years from now is of less importance than obtaining food, clothing, and shelter today. This program demonstrates that services such as transportation and child care are important elements of care for this population. Culturally sensitive recruitment strategies, personalized care, and vigilant follow-up are important requisites to breast cancer screening programs. A multidisciplinary team effort is critical in bringing together the elements required for a successful screening program. PMID- 9341353 TI - Cancer conferences. Can they be improved? AB - PURPOSE: Cancer conferences are required for hospital cancer program approval by the American College of Surgeons. These conferences are important educational and clinical opportunities and can influence the management of patients with cancer. Nationally, they represent an enormous expenditure of time and effort by physicians, associated healthcare personnel, and tumor registrars. The educational aspects of cancer conferences have been previously reviewed. The purpose of this investigation was to evaluate the clinical aspects of cancer conferences. DESCRIPTION OF STUDY: A questionnaire, inquiring about various elements of cases presented at six consecutive cancer conferences, was sent to 93 Illinois hospitals. These elements included presentation at conference (presenter, time of presentation), clinical aspects (symptoms, history, physical examination, and laboratory tests), pathology (TNM stage and markers), therapeutic options, and quality-of-life issues. RESULTS: The person (or persons) presenting the case was most frequently the attending physician (n = 805, 52%); followed by the pathologist (n = 427, 28%); the cancer committee chairperson (n = 318, 21%); the resident (n = 138, 9%); and other members of the multidisciplinary healthcare team (n = 525, 34%), such as the nurse practitioner or radiation therapist. Of the 1547 cases reviewed, history, physical examination, and diagnostic tests were discussed in 93%, 91%, and 93% of conference presentations, respectively. However, staging by the required TNM system, tumor markers, and quality-of-life issues were discussed in only 28%, 34%, and 38% of presentations, respectively. CLINICAL IMPLICATIONS: Although clinical characteristics were adequately documented and discussed at the cancer conferences studied, other important parameters, such as TNM staging, tumor markers, and quality-of-life issues, were less often discussed. The former topic frequencies are expected, the latter unacceptable. Although cancer conferences currently enhance patient care, these findings indicate that there is potential for improvement through discussion of TNM staging, tumor markers, and quality of life. PMID- 9341355 TI - Dietary changes after breast cancer. What should we recommend? PMID- 9341354 TI - Quality of life and breast cancer survivors. Psychosocial and treatment issues. AB - PURPOSE: This study was conducted to determine the relationship between social support, extent of breast cancer surgery, length of time since surgery, geographic location, and overall quality of life (QOL) in breast cancer survivors. Additionally, the motivational factors for survivors to volunteer in a community rehabilitation program were assessed. DESCRIPTION OF STUDY: A convenience sample of 134 survivors who had undergone mastectomies were invited to participate. Study packets including an information letter, a demographic sheet, and Ferrans and Powers Quality of Life-Cancer Version questionnaire were mailed. The return of completed forms was considered consent. RESULTS: The sample (N = 100) consisted mostly of married women (75%) who had surgery a mean of 14 years previously. Findings indicated that there is no correlation between marital status and the number or type of support persons and overall QOL. Interestingly, unmarried women were found to have a better perceived QOL than married women, although the difference was not statistically significant. There was no significant difference between extent of surgery, length of time since surgery, and geographic location with overall QOL. Thematic analysis of motivation to volunteer revealed an underlying theme of helping with sharing knowledge/providing information and giving emotional support for all age groups. Other themes were personal gain; giving back to the program, others, and God; and assisting others in avoiding a negative experience. CLINICAL IMPLICATIONS: Quality of life is a dynamic, multifaceted process through which perceptions, viewpoints, and behaviors change as a result of the various experiences throughout the survival period. These findings demonstrate that social support plays a vital role in promoting overall QOL in breast cancer survivors. The development of supportive behaviors by healthcare providers and Reach for Recovery volunteers is essential in providing this social support for breast cancer survivors. Growth in understanding and relieving the psychological stress that new cancer survivors endure is an area that warrants particular attention. Additionally, these participants indicated their need and willingness to share their experiences. Support is needed to provide survivors with the opportunity to function as volunteers. Program developers and evaluators must work together to ensure this support on a consistent basis. PMID- 9341356 TI - Life insurance policies and the terminally ill. PMID- 9341357 TI - Dolasetron. A new 5-hydroxytryptamine3 receptor antagonist. PMID- 9341406 TI - HIV transmission via breastfeeding: reflections on the issues. PMID- 9341407 TI - The power of support groups: influence on infant feeding trends in New Zealand. AB - By outlining infant feeding trends and the essential characteristics of support groups, this paper reveals how important such groups and their development have been in shaping the history of infant feeding in twentieth-century New Zealand. The paper draws, in particular, on the histories, growth, and influence of the Royal New Zealand Plunket Society, Parents Centre New Zealand, and La Leche League New Zealand. It demonstrates the importance of such middle-class groups in changing practices and attitudes within society. In general, this paper is a call for recognition of the importance of lay support groups in the improvement of health outcomes. PMID- 9341408 TI - Legal commentary: cyberspace and the lactation consultant. PMID- 9341409 TI - Nipple shields: LCs not looking for quick fix. PMID- 9341410 TI - Nipple shields: just another tool. PMID- 9341411 TI - Comparing breastfeeding and breast pumps using a computer model. AB - There is a role for computer models in increasing the understanding of milk extraction from the human teat. A computer model can be used to investigate aspects of extracting milk from the human teat which are not feasible using clinical experiments. In this paper, the behavior of the human teat during an infant suckling and with the use of a breast pump is modeled. The model is used to (1) identify the role of suction and the peristaltic motion of the tongue during suckling and (2) compare the volume of milk extracted by an infant breastfeeding with that obtained using a breast pump. Infants use a peristaltic motion of the tongue, along with some suction, to extract milk. Breast pumps use a cyclic pattern of suction only. In the model, the human teat is represented as a cylindrical porous elastic material saturated with fluid. We mimic an infant suckling by imposing both suction and a peristaltic force in the computer model of the human teat. This is compared to the effect of suction only, which models the action of breast pumps. The results demonstrate that there is an optimal time to apply the peristaltic force during the suction cycle which will increase the milk volume. The model and results may be of use in the future design of effective breast pumps. PMID- 9341412 TI - Do consumer infant feeding publications and products available in physicians' offices protect, promote, and support breastfeeding? AB - The purpose of this study was to determine if consumer infant feeding publications and products distributed by physicians' offices protect, promote, and support breastfeeding. A total of 127 physician office practices completed a mailed questionnaire that measured the types of print and nonprint materials available and policies and practices regarding these resources. Commercially produced pamphlets were available in 114 (90%) of the offices surveyed, and were twice as likely to be routinely distributed as pamphlets from nonprofit agencies and government. Many publications contained outdated recommendations about breastfeeding; the most accurate publications were available in only 29 (23%) of practices surveyed. Magazines contravening the WHO Code were widely available (91 offices; 72%) and routinely distributed (58 offices; 46%). One hundred one offices (80%) accepted free formula and 48 (38%) routinely distributed it. Few offices had a policy (n = 25; 20%) or criteria (n = 17; 13%) for selecting infant feeding resources. Those with policies were less likely to distribute commercial pamphlets. In the majority of offices surveyed, physicians' offices accepted and routinely distributed publications and products which do not "protect, promote, and support" breastfeeding. Offices are encouraged to have policies guiding the distribution of infant feeding materials. PMID- 9341414 TI - Employer attitudes toward breastfeeding in the workplace. AB - A descriptive, exploratory study of 69 male and female employers was done in a small rural community to determine their attitudes toward breastfeeding or expressing milk in the workplace. Business variables, such as experience working with women who have breastfed and knowledge of other businesses who have employed breastfeeding women, appeared to be better predictors of a positive level of support toward breastfeeding in the workplace than personal attributes, such as age, education level, and personal history with a spouse or friend who breastfed. The health care provider needs to become instrumental in promoting breastfeeding in the workplace by focusing on the positive effects on the business and providing employers with successful examples of workplace breastfeeding programs. PMID- 9341413 TI - The effect of feeding glucose water to breastfeeding newborns on weight, body temperature, blood glucose, and breastfeeding duration. AB - In order to determine the effect of feeding glucose water on breastfeeding newborns, we randomly distributed 180 normal newborns into two groups: a glucose water group (GW), fed 5% glucose solution during the first 3 days of life in addition to being breastfed; and an exclusively breastfed nonglucose water group (NGW). The following data were evaluated: weight at 6, 12, 24, 48, and 72 hours of life; temperature during the first 72 hours of life; serum glucose level at 6, 12, 24, and 48 hours; total duration of breastfeeding and age at introduction of infant formulas. In the NGW, there was a greater weight loss at 48 hours but not at 72 hours, temperatures higher than 37.5 degrees C were more frequent, and the mean serum glucose levels at 6, 12, and 24 hours were lower. This group also had more serum glucose level determinations under 2.2 mmol/l (40 mg/dL). However, no infants exhibited hypoglycemic symptoms. Infants in the GW received twice as many formulas during the first month and had a shorter duration of any breastfeeding. Our results suggest that the suppression of feedings with glucose water in the first days of life increases the probability of successful breastfeeding. However, infants who do not receive glucose water in the first few days of life may require greater supervision and close monitoring of blood glucose and body temperature, particularly in the first 24 hours of life. PMID- 9341415 TI - Dietitians in breastfeeding management: an untapped resource in the hospital. AB - Maternity care nurses were surveyed to gather data on the state of breastfeeding in Utah hospitals. A questionnaire was distributed to labor and delivery, well baby nursery, and postpartum nurses at 18 hospitals representing 86% of births in the state. The results indicated that of dietitians, physicians, and nurses, dietitians were the health care professionals least likely to provide breastfeeding information and assistance. PMID- 9341416 TI - Methadone levels in breast milk. AB - Two case reports of breastfeeding mothers on high doses of methadone and a literature review reveal that minimal transmission of methadone into breast milk occurs regardless of the mother's methadone dose. The current American Academy of Pediatrics recommendations that only women in drug treatment programs on less than 20 mg/day of methadone be advised to breastfeed should be reconsidered. PMID- 9341417 TI - Breastfeeding the infant with PKU. AB - Breastfeeding is possible with children who have phenylketonuria. Based on the decisions made by the breastfeeding mother and the infant's physician, there are multiple methods of management which can be utilized to provide breast milk for the infant with PKU. This article describes some of the methods that have been used to manage the treatment of phenylketonuria when a mother chooses to breastfeed. PMID- 9341419 TI - How North American donor milk banks operate: results of a survey: Part 2. PMID- 9341418 TI - A guide to references on drugs and breastfeeding. AB - Many of the above references will provide the necessary information to determine the safety of drugs during breastfeeding. Because interpretation of existing data varies in these references, it is prudent to consult with more than one resource. References should be routinely updated every 1 to 2 years as new drugs and literature are made available. PMID- 9341420 TI - Recent references. PMID- 9341422 TI - The holistic management of pressure sores in terminal illness. PMID- 9341421 TI - Local injection of phentolamine to treat digital ischaemic necrosis in Raynaud's syndrome. PMID- 9341423 TI - Case report challenge. PMID- 9341424 TI - Special equipment in nursing homes. PMID- 9341425 TI - Two bandaging systems. PMID- 9341426 TI - Two bandaging systems. PMID- 9341427 TI - Selection of dressings for diabetic foot ulcers. PMID- 9341428 TI - The leg ulcer resource team: a standard of care. AB - A leg ulcer resource team was formed to introduce and implement a standard of care for patients with leg ulcers. An audit of the standard following implementation showed that 82% of ulcers had been classified correctly, diagnostic tools had been used in 78% of cases and a clear rationale for care given in 86%. However, accurate identification of patient needs in the care plan occurred in only 39% of cases, and only 60% of care plans had ulcer size and site recorded. Patient education was also a problem--25% of patients did not know what type of ulcer they had and only 43% could describe care of their ulcer or identify risk factors associated with leg ulceration. PMID- 9341429 TI - Managing animal bite wounds. PMID- 9341430 TI - A comparison of two dressings in the management of chronic wounds. AB - A hydropolymer dressing (Tielle) and a hydrocolloid dressing (Granuflex) were compared in a randomised controlled clinical study involving 100 patients with leg ulcers and 99 patients with pressure sores in the community. Statistically significant differences in favour of the hydropolymer dressing were detected for dressing leakage and odour production, but no statistically significant differences were recorded in the number of patients with either leg ulcers or pressure sores who healed in each treatment group. PMID- 9341432 TI - The development of pressure sores in patients with spinal cord injury. PMID- 9341431 TI - Social support for elderly patients with chronic wounds. AB - Few attempts have been made to measure the social support received by elderly patients with chronic wounds. To focus research on these issues, an established model integrating the various roles played by social support in the adaptation of patients to stressful situations was applied. Two questionnaires were used to measure perceived social support and coping in a sample of patients with leg ulcers (N = 15, mean age 70.4 years) or diabetic foot ulcers (N = 15, mean age 63.6 years) at two time-points over a four-month period. The results indicate that there were no statistical differences between the groups. The overall levels of social support were low, with emotional support recorded most frequently. The standardised scores for types of coping indicate no unusual patterns, although the scores for logical analysis were low. However, there was considerable variation in the types of coping strategies used by individuals. PMID- 9341433 TI - Pressure sore treatment. 1. Accurate data collection. PMID- 9341434 TI - Pressure sore treatment. 2. Evidence of effectiveness. PMID- 9341468 TI - "Where have all the leaders gone?". PMID- 9341435 TI - Pressure sore treatment. 3. Clinical guidelines. PMID- 9341469 TI - Health link. PMID- 9341470 TI - A visit to Trinidad & Tobago. AB - Trinidad & Tobago are the two most southerly of the Caribbean islands off the coast of Venezuela. Trinidad is industrial, having natural deposits of oil, gas and bitumen, while Tobago is a tropical holiday island, sometimes known as Robinson Crusoe Island. The population of 1.3 million represents races from Africa, India, Europe and Asia, who live and work together harmoniously; Trinidadian women are renowned for their beauty, resulting from this rich mix. The centre of Trinidad is heavily forested and-apart from the capital, Port-of Spain-the towns are small and rural in character. Fruit grows abundantly, especially mangoes, bananas, pineapples, citrus fruits, paw-paw and coconuts. PMID- 9341471 TI - Contraceptive advice for teenagers in Finland. PMID- 9341472 TI - Meeting report. "Pregnancy": the inaugural conference of the Association for Infant Mental Health. PMID- 9341473 TI - Opinion. Clinical support is essential for midwifery students. PMID- 9341474 TI - Risks of cot death. PMID- 9341541 TI - Promoting humanity. PMID- 9341542 TI - The effectiveness of massage in preventing pressure sores: a literature review. AB - The prevention of pressure sores is a major concern of rehabilitation nurses. Through the years, several methods have been used to prevent pressure sores. One of the most commonly used methods is massage of bony prominences and pressure areas. However, according to most contemporary clinical guidelines, massage should be avoided. This article analyzes the extent to which these guidelines are based on research findings through a literature review on the effectiveness of massage in the prevention of pressure sores. The results of the studies that were analyzed led to the conclusion that massage as therapy for preventing pressure sores in subjects at risk for developing them is not recommended. PMID- 9341543 TI - Interdisciplinary documentation of patient education: how collaboration can effect change. AB - Changing healthcare trends are affecting all healthcare providers today, including those at Hillside Rehabilitation Hospital. Computerizing the interdisciplinary documentation of patient information was one change Hillside's leaders implemented to remain competitive and cost-effective. The benefits of moving from a handwritten documentation system to a computerized system were many. However, unforeseen difficulties with retrieving data became evident because the system was not developed to accommodate information about patient education. Through interdisciplinary collaboration, staff identified areas for improvement and made changes to the documentation process. They developed a "traveling card" for documenting patient education and then met a new set of challenges. The new documentation system met departmental and Joint Commission on Accreditation of Healthcare Organizations requirements for comprehensive patient education and documentation. PMID- 9341544 TI - The rehabilitation nurse in the home care setting: care of the client with HIV or AIDS. AB - People with HIV or AIDS who are experiencing pain, fatigue, sexual dysfunction, bowel and bladder dysfunction, and self-care deficits are being cared for by rehabilitation nurses in the home setting. The home care rehabilitation nurse provides instruction and care to clients, their families, and caregivers regarding physical manifestations of the disease and issues such as the importance of involving the client in household activities and activities of daily living. In addition to working with an interdisciplinary team to meet clients' needs, home care rehabilitation nurses work and consult with the generalist nursing staff to offer recommendations about rehabilitation nursing care for clients with HIV or AIDS. PMID- 9341545 TI - Stroke rehabilitation: assessing stroke survivors' long-term learning needs. AB - What are the long-term learning needs of stroke survivors who must adapt to living with stroke-related disabilities? A broad-based educational needs assessment was conducted to find answers to this question and to gain insight into the concerns and challenges facing stroke survivors who have returned to living in the community. Stroke survivors, family members, and rehabilitation healthcare professionals were surveyed; positive responses strongly indicated that there was interest in learning more about the selected topics. Although there was congruence across all groups, healthcare professionals indicated that they felt more strongly than either patients or patients' family members that stroke survivors need to learn more about the facts and statistics of strokes. Stroke survivors and family members indicated a stronger interest in learning about complementary therapies such as massage, acupuncture, and the role of food and vitamins. Time since stroke was an influencing factor for both stroke survivors and family members in their responses to how strongly they wanted to learn about various topics. The findings from this needs assessment formed the basis for an educational course that uses a holistic approach to address the physical, emotional, social, economic, and spiritual aspects of adapting to living with stroke-related disabilities. PMID- 9341546 TI - Motivation and the coping process of adults with disabilities: a qualitative study. AB - Adults with disabilities who have completed rehabilitation programs and have returned to active lifestyles are experts on the importance of motivation after an injury or an illness. This qualitative descriptive study was conducted with 9 men and 3 women who had completed a spinal cord injury rehabilitation program at a rehabilitation hospital. The subjects were asked two questions: What helped motivate you during rehabilitation to return to an active, productive life? and How did rehabilitation nurses and staff assist you with that process? An analysis of the interviews revealed five motivational categories--independence, education, socialization, self-esteem, and realization--within the specific themes of nursing and healthcare interventions. Gaining insight into the motivation of adults who have coped with disabilities effectively can help rehabilitation nurses determine how they can promote the motivation that clients need to achieve a quality lifestyle. PMID- 9341547 TI - Ice therapy for pressure sores. PMID- 9341548 TI - Promoting skin integrity: an interdisciplinary challenge. AB - The process of standardizing protocols to promote skin integrity in an interdisciplinary setting has proved to be an ongoing process. The referral team, nurses, physical therapists, nutritionists, and physicians continue to provide standardized care. Through this interdisciplinary process, the core skin care team continues to refine and develop standards to reflect current research findings. As we work together and share our knowledge, we improve the quality of care for our patients. PMID- 9341549 TI - Success or tragedy? PMID- 9341562 TI - Analysis of fast single-joint and multijoint movements in cerebellar cortical atrophy: failure of L-hydroxytryptophan to improve cerebellar ataxia. PMID- 9341563 TI - Simple partial status epilepticus and antiglycolipid IgM antibodies: possible epilepsy of autoimmune origin. PMID- 9341564 TI - The aging brain. Limitations in our knowledge and future approaches. PMID- 9341565 TI - A neurologist's perspective on the aging brain. AB - I have worked as a neurologist in old age homes and chronic hospitals for 30 years, and over this period I have seen thousands of older patients with dementias and other degenerative neurological syndromes. A high proportion of these patients have steadily and sometimes rapidly advancing dementias of the type we now tend to call Alzheimer disease (AD) (although this term was used in a much narrower sense when I started seeing such patients in the mid-sixties). PMID- 9341566 TI - Oldest-old healthy brain function. The genomic potential. AB - There will be a remarkable increase in the number of people older than 85 years- the oldest-old--in the next 50 years. This age group is especially vulnerable to increasing disabilities, many of which are the result of the aging nervous system. Among these aging changes, the most devastating and likely to have the greatest impact on our society is the development of dementia. Since dementia is present in the majority of those who live to the current maximum of the human life span, this suggests that dementia is a normal aging event. Although the exact causes of this common cognitive failure in the oldest-old are not known, there is recent evidence from genetic studies of aging and Alzheimer disease to suggest that there are a number of susceptibility genes that may modify or delay the onset of late-life brain failure. These gene families form a natural target for devising strategies for significantly delaying the onset of late-life dementia. PMID- 9341567 TI - New therapies for stroke. AB - The treatment of patients with stroke and cerebrovascular disease has entered a new era. During the 1990s there has been a revolution in technology able to define quickly, safely, and accurately stroke pathophysiological characteristics and the cardiovascular lesions that cause stroke in individual patients. Advanced brain imaging with computed tomography, magnetic resonance imaging, and newer magnetic resonance modalities, including fluid attenuating inversion recovery imaging, diffusion, perfusion, functional magnetic resonance imaging, and magnetic resonance spectroscopy, show clinicians the localization, severity, and potential reversibility of ischemia. Vascular lesions can be defined using spiral computed tomographic angiography, magnetic resonance angiography, and extracranial and transcranial ultrasonography. Cardiac and aortic sources of stroke are now better studied using transesophageal echocardiography. More sophisticated hematologic testing gives new insights into the role of altered coagulability in causing or contributing to thromboembolism. Clinicians can now recognize the key data elements needed to logically treat brain ischemia, including the following: The nature, location, and severity of cardiac and cerebrovascular lesions. The mechanism by which these lesions cause ischemia- hypoperfusion? embolism? functional changes such as vasoconstriction? The cellular and serologic components of the blood that relate to coagulability, viscosity, and blood flow. The state of the brain--normal, reversibly ichemic ("stunned"), or infarcted. With these diagnostic advances have come new treatments, new ideas about treatment, and more and new information about conventional treatments. PMID- 9341568 TI - Genetic testing for Alzheimer disease. Practical and ethical issues. AB - The dissection of the heterogeneous genetics of the Alzheimer diseases (ADs) is currently more advanced than in any other common disease. Not only are 3 uncommon autosomal dominant mutation loci identified, but universally inherited susceptibility polymorphisms associated with risk and age of onset distributions for familial and "sporadic" AD are also confirmed. The utility of testing for mutations of the amyloid precursor protein and the presenilin 1 and presenilin 2 genes conforms to strategies in common use for rare mutations. The selection of patients with very early-onset AD, especially those with family histories of the disease, will increase the possibility of diagnosis. All testing should be performed using recommended counseling procedures. Apolipoprotein E (ApoE) susceptibility polymorphisms are genetic risk factors but do not allow prediction of the age of onset of AD for cognitively normal individuals. Recent large, collaborative studies have found that when ApoE genotyping is used sequentially in diagnosis following criteria-based evaluations, the specificity of early diagnosis is significantly increased. For patients clinically diagnosed with AD who carried an ApoE epsilon 4 allele, the positive predictive value was 94% and 97% in 2 multicenter collaborative series. The utility of ApoE genotyping is reviewed and recommendations for early use in diagnosis are explained. There are ethical, social, actuarial, and legal problems currently associated with genetic testing and these concerns are also discussed. PMID- 9341569 TI - Excitotoxic neurodegeneration in Alzheimer disease. New hypothesis and new therapeutic strategies. AB - Excessive activation of N-methyl D-aspartate (NMDA) receptors by endogenous glutamate (Glu) causes excitotoxic neuronal degeneration in acute central nervous system injury syndromes such as stroke and trauma. Early attempts to link NMDA receptor hyperactivity (NRHyper) to Alzheimer disease (AD) were stymied by evidence in 3 separate species (mice, rats, and monkeys) that, with advancing age, the NMDA receptor system becomes markedly hypoactive. While this would seem to argue against a role for NMDA receptors in AD, we have recently found in animal studies that, when the NMDA receptor system is rendered markedly hypoactive, a disinhibition syndrome is triggered in which low-grade chronic excitotoxic activity (fueled by acetylcholine and Glu) is unleashed that can cause a widespread pattern of neuronal degeneration resembling that seen in AD. Therefore, we postulate that NMDA receptor hypoactivity (NRHypo) associated with advancing age may have an important contributory role in AD and that the main difference between the aging AD brain and the aging "normal" brain is that a heavier burden of certain adjunctive risk factors may be present in the AD brain that promote the NRHypo state and increase the likelihood that widespread neurodegeneration will occur. PMID- 9341570 TI - Amyotrophic lateral sclerosis. Insights from genetics. AB - Amyotrophic lateral sclerosis (ALS) is among the most dire neurological diseases. The essential clinical feature of the disorder is relentless, lethal paralysis, usually beginning in midadult years. The disease is caused by a slow, progressive loss of motor neurons in the brain and spinal cord. It usually begins focally and then spreads. In most cases, there is concurrent involvement of corticospinal (upper) and spinal (lower) motor neurons, although in some instances the spinal motor neuron features predominate. Involvement of the spinal motor neurons produces muscle denervation of the affected muscles and fasciculations, followed by muscle atrophy. When corticospinal motor neurons degenerate, the weakness is accompanied by spasticity. The mean age at onset of ALS is 55 years; the mean duration is about 4 years. The incidence of new cases is approximately 1 per 100,000 population. The total number of cases is about 5 per 100,000 population. In the United States, it is estimated that there are 20,000 to 30,000 cases. About 10% of cases are inherited as an autosomal dominant trait; familial and sporadic ALS are clinically indistinguishable. PMID- 9341571 TI - Presidential judgment. The twenty-fifth amendment. PMID- 9341572 TI - Disability in US presidents report. Recommendations and commentaries by the Working Group. The Working Group on Presidential Disability. AB - If the president of the United States must decide within minutes how to respond to a dire emergency, its citizens expect him or her to be mentally competent and to act wisely. Because the presidency of the United States is now the world's most powerful office, should its incumbent become even temporarily unable to exercise good judgment, the consequences for the world could be unimaginably far reaching. PMID- 9341573 TI - The delay in reporting symptoms of carotid artery stenosis in an at-risk population. The Asymptomatic Carotid Atherosclerosis Study experience: a statement of concern regarding watchful waiting. AB - OBJECTIVE: To examine whether patients in the Asymptomatic Carotid Atherosclerosis Study reported symptoms of cerebral and retinal ischemia promptly to the investigating team. DESIGN: Cohort study within the Asymptomatic Carotid Atherosclerosis Study, a prospective, randomized, multicenter clinical trial, with a median follow-up time of 2.7 years. SETTING: Thirty-nine clinical sites across the United States and Canada. PATIENTS: Patients with asymptomatic carotid artery stenosis (> or = 60% reduction in diameter) who experienced either a transient ischemic attack (TIA) (n = 115) or stroke (n = 127) during the follow up period, as verified by an external committee. MAIN OUTCOME MEASURE: Proportion of patients who reported cerebrovascular symptoms to a study nurse or physician within 3 days of occurrence. RESULTS: Thirty-seven patients (32.2%) experiencing TIA and 57 (44.9%) experiencing stroke reported symptoms to the study staff within 3 days of onset. For TIA, there was a statistically significant inverse association between prompt reporting and the amount of time a patient was enrolled in the study before the event occurred (48% with TIA occurring within 6 months vs 9% with TIA after year 3; P = .04). For stroke, there was a statistically significant association between prompt reporting and treatment arm (56% for the surgical vs 38% for the medical group; P = .05). For either TIA or stroke, none of the other factors examined were significantly associated with prompt reporting. CONCLUSIONS: Despite extensive education and reinforcement, fewer than 40% of all first events were reported within 3 days and fewer than 25% were reported in less than 24 hours. Frequent outpatient evaluation of high-risk patients and careful review of symptoms is necessary to determine when asymptomatic carotid artery stenosis has become symptomatic to offer appropriate forms of therapy. PMID- 9341574 TI - Features, symptoms, and neurophysiological findings in stroke associated with hyperhomocysteinemia. AB - BACKGROUND: Hyperhomocysteinemia has been shown to be a mild independent risk factor for premature atherosclerosis, and there is evidence of an increased rate of peripheral vascular occlusive disease, myocardial infarction, and stroke. OBJECTIVE: To evaluate clinical, biochemical, and neurophysiological findings in patients with ischemic stroke with and without hyperhomocysteinemia. SUBJECTS: One hundred twenty-five consecutive patients with a history of stroke and 60 healthy control subjects. METHODS: Patients were divided into those with and those without hyperhomocysteinemia, which was defined as blood levels beyond the mean total plasma homocysteine level plus 2 SDs of the healthy control group. History, symptoms, cause, patterns of infarction, biochemical data, continuous and transcranial Doppler sonography, and event-related potentials were recorded in all patients. RESULTS: Twenty-seven patients had hyperhomocysteinemia. Compared with the 98 patients without hyperhomocysteinemia, they had an increased rate of hypertension (odds ratio, 3.5; 95% confidence interval, 1.0-12.6), an increased level of uric acid (P < .007), an increased hematocrit (P < .02), a higher rate of microangiopathy (odds ratio, 2.8; 95% confidence interval, 1.1 7.2), and a trend to a higher rate of multiple infarction. Furthermore, the P3 latency of the event-related potential was significantly increased in hyperhomocysteinemia (P < .004). CONCLUSIONS: Hyperhomocysteinemia is probably an independent risk factor for stroke, with a prevalence of about 20% in all patients with a history of stroke; however, additional factors (eg, hypertension, hyperuricemia) may have an enhancing effect. There are significant differences in stroke patterns between patients with and without hyperhomocysteinemia, with a higher rate of lesions typical of cerebral microangiopathy and a trend to multiple infarctions in the former. Impairment of cognitive processing as measured by visual event-related potential is more pronounced in hyperhomocysteinemia. PMID- 9341575 TI - Periventricular white matter lucencies in patients with lacunar stroke. A marker of too high or too low blood pressure? AB - BACKGROUND: Periventricular white matter lucencies (PWML) have been described in stroke patients with arterial hypertension, hypotensive episodes, or increased nocturnal fall of blood pressure (BP). As a result of these mixed factors, the relationship between PWML and BP remains unsettled and the appropriate management of arterial BP in stroke patients with PWML is unknown. OBJECTIVE: To clarify the relationship between PWML, arterial BP, and cerebral hemodynamics. DESIGN: Cohort study followed up 6 months after index stroke. SETTING: Referral center. PATIENTS: In 41 consecutive patients with first-ever lacunar infarction, the extent of PWML detected on brain magnetic resonance images was measured. Six months after stroke, BP values were monitored during a 24-hour period and transcranial Doppler examinations were performed at rest and following the administration of acetazolamide. MAIN OUTCOME MEASURES: Correlation of cerebral hemodynamics and BP values with the extent of PWML. RESULTS: The severity of PWML varied substantially among patients, suggesting that PWML and lacunar infarctions could be due to several different mechanisms. Older age, elevated awake systolic BP, increased cerebrovascular tone, and the interaction between history of heart disease and the lowest heart rate were the strongest independent predictors of the severity of PWML. Diastolic BP and the vasodilatory capacity of the resistance vessels did not predict the severity of PWML. CONCLUSIONS: Overall, PWML are markers of systolic damage in older lacunar stroke patients with stiffer arteries. In addition, hemodynamic failure may be relevant in patients with concomitant heart disease. PMID- 9341576 TI - Lack of association of a polymorphism in the low-density lipoprotein receptor related protein gene with Alzheimer disease. AB - BACKGROUND: The apolipoprotein E epsilon 4 (ApoE epsilon 4) allele is associated with an increased risk for development of Alzheimer disease (AD). We hypothesized that polymorphisms in proteins that interact with ApoE also might have an impact on the likelihood of AD developing. We examined a polymorphism in the gene for the low-density lipoprotein receptor-related protein (LRP), because LRP is a major ApoE receptor in the brain that also mediates binding and degradation of secreted Kunitz protease inhibitor forms of amyloid precursor protein. SUBJECTS AND DESIGN: The LRP genotypes in 2 groups were studied. The first group consisted of 130 patients with probable or definite AD (mean +/- SD age, 78.2 +/- 8.9 years) and 64 nondemented, control subjects (mean +/- SD age, 81.7 +/- 12.3 years) who were primarily the spouses of the patients. The second group consisted of individuals from a population-based, epidemiologic study, including 38 cognitively impaired individuals (mean +/- SD age, 79.9 +/- 4.1 years) and 93 nondemented controls (mean +/- SD age, 78.7 +/- 4.4 years). Finally, 22 brains with a neuropathological diagnosis of AD were evaluated for neuronal loss, beta amyloid deposition, and neurofibrillary tangle number and compared with the LRP genotype. RESULTS: No genetic disequilibrium in LRP allele frequencies between controls and patients with AD or cognitive impairment was observed. No interaction between the ApoE epsilon 4 and LRP genotypes was observed in patients with AD. Moreover, the LRP genotype did not correlate with degree of neuronal loss, beta-amyloid deposition, or neurofibrillary tangle number in individuals examined using quantitative neuropathological techniques. CONCLUSION: This LRP gene polymorphism is not linked with the pathophysiological changes of AD. PMID- 9341577 TI - Bilateral temporal lobe volume reduction parallels cognitive impairment in progressive aphasia. AB - BACKGROUND: Patients with isolated aphasia in the absence of other cognitive abnormalities have been the focus of several studies during the past decade. It has been called primary progressive aphasia (PPA), and the typical features of this syndrome are marked atrophy of the left temporal lobe according to the radiological examination and a language disorder as the initial symptom. In previous studies of PPA, the selection of the patients was based mainly on linguistic symptoms. Now, when computed tomography or magnetic resonance imaging scans are part of the routine investigation of cognitive impairment and suspected dementia, the patients with lobar atrophy will be found at an earlier stage. In the present study, we used a new approach and defined the study group by selecting patients with obvious left temporal lobe atrophy, assessed by MRI, and we referred to them as patients with temporal lobe atrophy (TLA). OBJECTIVE: To identify the features that distinguish TLA from other primary neurodegenerative disorders. PATIENTS: Six patients with TLA were compared with patients with Alzheimer disease (AD), patients with frontal lobe dementia (FLD), and healthy control subjects. METHODS: The investigations included magnetic resonance imaging volumetry, single photon emission computed tomography, and neuropsychologic and linguistic evaluations. RESULTS: In the TLA group, the mean volume of the left temporal lobe was 35% smaller than the right, while in the AD and FLD groups, the atrophy was symmetrical and bilateral. In the TLA group, the absolute volumes of the temporal lobes were significantly smaller on the left side compared with the AD and FLD groups, whereas there was no difference on the right side. The cerebral blood flow pattern in TLA was asymmetric and differed from that in the other study groups. All patients with TLA had a history of progressive Wernicke type aphasia, ranging from 2 to 6 years. They showed primary verbal memory impairment but had preserved visuospatial functions. The clinical condition of all patients with TLA deteriorated during the study period; severe aphasia developed, and the patients exhibited signs of frontal lobe dysfunction. Serial volumetric measurements in 4 of 6 patients showed an annual 8% to 9% decrease of both left and right temporal lobes. CONCLUSIONS: The initial marked asymmetry in cognitive function found in patients with TLA contrasts with the general decline found in patients with AD. The bilateral degenerative process evident in patients with TLA paralleled the clinical deterioration, indicating TLA to be a non-AD lobar atrophy that develops into generalized cognitive dysfunction and dementia. PMID- 9341578 TI - Gut response. Therapy with ingested immunomodulatory proteins. AB - The recent negative results of the 2-year phase III trial of enteral myelin (Myloral), a preparation of bovine myelin given orally in multiple sclerosis (MS), warn that the efficacy of oral tolerization (ie, inducing hyporesponsiveness to a specific autoantigen) and the attractiveness of oral desensitization in animal models of MS may not translate into clinical practice. However, such failure should not deter further examination of the mouth, an immunologically interesting and patient-friendly route for immunomodulatory proteins. PMID- 9341579 TI - Research and training on the newly dead. PMID- 9341580 TI - Histologic occurrence of polystyrene sulfonates. PMID- 9341581 TI - Changes in emergency department turnaround time performance from 1990 to 1993. A comparison of two College of American Pathologists Q-probes studies. AB - OBJECTIVE: To compare the results of a 1990 College of American Pathologists Q Probes Emergency Department (ED) turnaround time (TAT) study with a similar study done in 1993 and to identify factors associated with TAT improvement. DESIGN: Participants gathered data over a 4-week period on the various times of day associated with the ordering, specimen-collection, laboratory-receipt, and result reporting stages of stat tests for potassium and hemoglobin levels, using a mail in questionnaire that also included practice parameter questions. PARTICIPANTS: Laboratories enrolled in the 1990 College of American Pathologists Q-Probes study on ED TAT and laboratories enrolled in the 1993 program ED TAT study. MAIN OUTCOME MEASURES: Components associated with shorter ED TAT and, for those participating in both the 1990 and 1993 studies, comparison with previous results. RESULTS: Six hundred fifteen hospital laboratories returned data on up to 43,521 hemoglobin and 41,989 potassium specimens. Half of these laboratories collected and reported 90% of their ED potassium results in 53 minutes or less, compared to 61 minutes or less in 1990, and reduced the corresponding median collection-to-reporting TAT for 90% of hemoglobin results from 46 minutes or less to 39 minutes or less. The fastest 10% of laboratories showed interlaboratory median order-to-report TATs of 36 and 50 minutes for potassium and hemoglobin tests, respectively. Comparisons of TATs from 277 laboratories with 1990 and 1993 data were possible. Components found to contribute statistically to improvement of ED TAT between 1990 and 1993 were laboratory control of specimen handling, rapid transport time, and monitoring. Active monitoring was particularly important when the laboratory did not control the specimen-handling process. CONCLUSIONS: Laboratories improved their control of ED TAT from 1990 to 1993 and reduced the number of TAT events exceeding 70 minutes. Internally set TAT goals, however, were not met most of the time. PMID- 9341582 TI - Protocol for the examination of specimens removed from patients with gastrointestinal lymphoma. A basis for checklists. The Cancer Committee, College of American Pathologists, and the Task Force for Protocols on the Examination of Specimens From Patients With Gastrointestinal Lymphoma. PMID- 9341583 TI - Fetal hemoglobin levels in sudden infant death syndrome. AB - OBJECTIVE: The aim of this study was to determine and compare fetal hemoglobin levels from infants dying of the sudden infant death syndrome (SIDS) with aged matched control infants dying of other causes. Similar previous studies have reported both elevated and normal levels of fetal hemoglobin in whole blood samples from infants dying of SIDS. DESIGN: Triton-acid-urea gel electrophoresis and densitometry were used to determine fetal hemoglobin levels in postmortem whole blood samples from infants dying of SIDS and from appropriately age-matched control infants. Whole blood samples were analyzed blindly and matched for postgestational age. Infant ages at death ranged from birth to less than 1 year. MAIN OUTCOME MEASURES: Fetal hemoglobin in whole blood from infants dying of SIDS and control infants. RESULTS: During the period of postnatal development most associated with SIDS cases (2 to 6 months after birth), fetal hemoglobin levels were found to be significantly elevated in postmortem whole blood samples from SIDS infants compared with gestational age-matched control infants dying of causes other than SIDS. CONCLUSION: We conclude that levels of fetal hemoglobin are elevated in postmortem whole blood of SIDS infants compared with controls. Furthermore, the apparent conflict in the literature regarding fetal hemoglobin levels in SIDS infants and controls is most likely due to variability in the control data of some studies. PMID- 9341584 TI - Intra-abdominal lymphangiomas in children and adults. Assessment of proliferative activity. AB - OBJECTIVE: Intra-abdominal lymphangiomas are rare in children and even more exceptional in adults. Because these lesions occasionally progressively enlarge, we analyzed seven adult and four pediatric cases for evidence of proliferative activity. DESIGN: Immunohistochemical analysis was performed retrospectively on representative tissue sections using antibodies to the following antigens: Ki-67, proliferating cell nuclear antigen, and p53 gene product (eight cases). DNA ploidy was examined in five cases. PATIENTS: The study group consisted of seven adult women (aged 24 to 73 years), a 3.5-year-old girl, and two boys, aged 3.5 and 9 years, the last with a recurrence at age 15. The lymphangiomas ranged from 1.7 to 23 cm in maximum size. RESULTS: Ranges of percentages of cells staining for proliferating cell nuclear antigen, Ki-67, and p53 were similar between the pediatric and adult cases. Antibody to Ki-67 stained from 0.5% to 17% of the stromal and endothelial components of the lymphangiomas. Proliferating cell nuclear antigen activity was noted in 16% to 52% of lesional cells. Reactivity was noted almost exclusively in areas of inflammation and fibroplasia. For comparison, 10% to 50% of intermixed lymphocytes stained for Ki-67 and proliferating cell nuclear antigen. There was no labeling with p53. DNA content was uniformly diploid. CONCLUSIONS: The scant staining for Ki-67 in the majority of the lesions, combined with proliferative rates that were only focally elevated, suggests that lymphangiomas in children and adults are quiescent lesions whose enlargement is due to engorgement by chyle and localized secondary inflammation rather than primary tumoral growth. PMID- 9341585 TI - Sensitivity and specificity of triphenyl tetrazolium chloride in the gross diagnosis of acute myocardial infarcts. AB - OBJECTIVE: Myocardial infarction is the most common cause of sudden death in the United States. However, the identification of early myocardial infarcts at necropsy is frequently difficult, since unequivocal gross changes of infarcts do not become apparent for 24 to 48 hours following myocardial ischemic injury. The use of dyes, such as nitro-blue tetrazolium and 2,3,5 triphenyl tetrazolium chloride (TTC), that identify the dehydrogenase-deficient infarcted myocardium has been shown, largely by animal studies, to be very helpful in the macroscopic diagnosis of such cases. Such animal studies could not be directly extrapolated to human autopsy studies, however, because of the assumption that autolysis may invalidate the histochemical assessment of myocardial enzymatic changes after death. This study was carried out to evaluate the sensitivity and specificity of TTC in the gross diagnosis of acute myocardial infarction in the human population. DESIGN: The TTC stain reactions were correlated with histologic findings in the hearts of 638 consecutive adult autopsies. RESULTS: Of the 638 hearts examined by TTC, 174 hearts stained positive for acute infarction; histology confirmed myocardial infarction in 140 hearts. Histologic examination revealed acute infarcts in 41 of the remaining 464 cases that had stained negatively with TTC. The use of TTC in the macroscopic diagnosis of acute myocardial infarcts in the human population was found to have a diagnostic sensitivity of 77.4% and a specificity of 92.6%. The predictive value of a positive test was 80.5%, and that of a negative test was 91.2%. CONCLUSIONS: The overall diagnostic efficiency of the TTC test (ie, number of patients correctly identified) was 88%. These results show that the TTC test is a reliable, sensitive, and specific adjunct in the examination of the human heart at necropsy. PMID- 9341586 TI - Comparison of atherosclerosis in alaska Natives and nonnatives. AB - OBJECTIVE: To test the hypothesis that Alaska Natives have fewer atherosclerotic lesions in the coronary arteries and aorta than nonnative Alaska residents. DESIGN: Systematic standardized collection and evaluation of coronary arteries and aortas collected at autopsy. SETTING: Forensic autopsy service in Alaska. SUBJECTS: One hundred thirty Alaska Natives and 115 Alaska nonnatives who underwent forensic autopsy between February 1989 and December 1993. INTERVENTION: None. MAIN OUTCOME MEASURES: Prevalence and extent of atherosclerotic lesions in the aortas and coronary arteries in both populations studied. RESULTS: Alaska Natives had significantly lower prevalence and extent of raised atherosclerotic lesions in the abdominal aorta and coronary arteries than nonnative Alaska residents. CONCLUSIONS: Differences in coronary heart disease mortality between Alaska Natives and nonnatives are, at least in part, the result of fewer atherosclerotic lesions in Alaska Natives. PMID- 9341587 TI - The implantable cardioverter-defibrillator. A potential hazard for autopsy pathologists. AB - The implantable cardioverter-defibrillator (ICD) is an implantable electronic device that has been proven to be safe and effective in treating various malignant tachyarrhythmias in susceptible individuals. As the use of ICDs becomes more widespread, more individuals with the implanted devices will be encountered at autopsy. Manipulation of an activated ICD can result in electrical shock. To avoid injury, pathologists must be properly prepared to deal with bodies containing activated ICDs. These devices can also provide valuable information that may be helpful in determining the cause and mechanism of death. Herein, we present information regarding the appropriate guidelines and safeguards for pathologists confronted with an activated ICD. PMID- 9341590 TI - Necrobiosis lipoidica in a diabetic intravenous drug user. AB - Necrobiosis lipoidica occurs most commonly on the lower extremities in patients with diabetes; lesions typically occur in the pretibial region. Although the pathogenesis of necrobiosis lipoidica remains unclear, both external trauma and vascular damage have been proposed as contributing to the development of this disorder. We report polarizable foreign material in small blood vessels and multinucleated histiocytes in lesions of necrobiosis lipoidica. This phenomenon occurred in a patient who was both diabetic and an intravenous drug user. PMID- 9341589 TI - The effect of smoking on the serum ionized magnesium concentration is method dependent. AB - OBJECTIVE: To investigate the effect of smoking on serum ionized magnesium concentration ([Mg2+]) determined by the NOVA and AVL Mg ion-selective electrodes (Mg ISEs). METHODS: Subjects were apparently healthy smokers (n = 30) and nonsmokers (n = 30). We determined NOVA and AVL [Mg2+] in their serum and in test solutions containing compounds increased by smoking. We also determined subjects' white blood cell and differential counts. RESULTS: For smokers, the mean values for NOVA and AVL [Mg2+] differed significantly (0.41 vs 0.52 mmol/L, respectively). We found a significant intramethod difference in NOVA [Mg2+] (0.11 mmol/L, P < .0001) between smokers and nonsmokers. A dose-dependent decrease in NOVA [Mg2+] was observed with an increase in cigarettes/day. NOVA [Mg2+] inversely correlated with white blood cell counts. There was no interference by the test compounds with either Mg ISE. CONCLUSION: Smoking may induce a serum factor, possibly related to white blood cells, that negatively interferes with the response of the NOVA Mg ISE. PMID- 9341588 TI - Renal chromophobe cell carcinoma and oncocytoma. A comparative morphologic, histochemical, and immunohistochemical study of 124 cases. AB - BACKGROUND: Renal oncocytoma has several features that overlap with other renal neoplasms, including the eosinophilic subtype of chromophobe cell carcinoma. In fact, strict criteria for renal oncocytoma have not been well defined and remain a matter of controversy. Ultrastructural studies or sophisticated methods such as flow cytometry and cytogenetic techniques can be of great use in distinguishing the two tumors, but are difficult to propose as routine methods because of their limited availability. OBJECTIVE: To further characterize the histologic criteria of these tumors, we undertook a retrospective study to define the utility of routinely available histochemical and immunohistochemical techniques. DESIGN AND SETTING: Twenty-one cases of chromophobe cell carcinoma, eosinophilic subtype, and 103 cases of oncocytoma were tested with histochemical (Perls, periodic acid Schiff, and Hale's colloidal iron) and immunohistochemical (peanut agglutinin antigen and UEA-1 for lectins; cytokeratin KL1, epithelial membrane antigen, vimentin, S100 protein, and lysozyme) staining. RESULTS: The antibodies tested and the histochemical staining using Hale's colloidal iron allowed eosinophilic chromophobe cell carcinoma to be distinguished by its characteristic reaction pattern. Seventy-six percent of the chromophobe cell carcinomas showed a microvacuolated pattern, and 89% of the renal oncocytomas showed an apical positivity with Hale's colloidal iron staining (P < .01). Peripheral cell accentuation reactivity for cytokeratin KL1 was observed in 66% of the chromophobe cell carcinoma cases, and apical cytoplasmic positivity was observed in 37% of the renal oncocytoma cases (P = .01). Significant patterns were observed with anti-epithelial membrane antigen and anti-peanut agglutinin antigen antibodies (P = .05 and P = .01, respectively). Positive reactions for vimentin, S100 protein, lysozyme, and UEA-1 were not significant characteristics. CONCLUSION: Our study demonstrated that a precise morphologic description associated with simple histochemical and immunohistochemical techniques provides sufficient criteria for a high level of discrimination between the eosinophilic subtype of chromophobe cell carcinoma and renal oncocytoma. PMID- 9341591 TI - Primary low-grade T-helper cell testicular lymphoma. AB - Non-Hodgkin's lymphoma is the most common primary testicular neoplasm of older men but is rare in young men. The vast majority of primary testicular lymphomas are intermediate- to high-grade lymphomas. Peripheral T-cell lymphomas of the testes are rare; most are large cell lymphomas. We describe an unusual case of a primary small lymphocytic, CD4+, peripheral T-cell lymphoma that presented as a Staphylococcus aureus scrotal abscess in a 30-year-old male. Clonal rearrangement of the T-cell receptor beta gene was demonstrated by Southern analysis. To our knowledge, low-grade, peripheral T-cell lymphoma of the testes has not been previously reported. PMID- 9341593 TI - Anaplastic carcinoma showing rhabdoid features combined with mucinous cystadenocarcinoma of the pancreas. AB - A 52-year-old Japanese woman presented with complaints of back pain, loss of appetite, and weight loss; her diagnosis was a mass in the pancreatic tail. A distal pancreatectomy combined with lymph node dissection was performed. The tumor measured 10.0 x 9.5 x 8.0 cm and consisted of a cystic mass and a solid area. Histologically, the cystic mass represented a typical mucinous cystadenocarcinoma, whereas the solid portion was composed of anaplastic tumor cells with round eosinophilic intracytoplasmic inclusions, as seen in malignant rhabdoid tumor of the kidney. The hyaline-like intracytoplasmic inclusions were weakly positive for periodic acid-Schiff and were resistant to diastase. There was a gradual transition between the mucinous cystadenocarcinoma and the anaplastic carcinoma showing rhabdoid features. Additionally, the tumor cells of the anaplastic carcinoma were immunoreactive to both epithelial membrane antigen and vimentin. These findings suggest that the anaplastic carcinoma with rhabdoid features developed from the mucinous cystadenocarcinoma of the pancreas. PMID- 9341592 TI - Clear cell carcinoid tumor of stomach. A variant mimicking gastric xanthelasma. AB - We report a rare case of gastric carcinoid composed entirely of clear cells with foamy cytoplasm. The patient was a 75-year-old Hispanic woman with atrophic fundic (oxyntic) gland gastritis leading to hypochlorhydria, secondary hypergastrinemia, endocrine cell hyperplasia, and multiple gastric carcinoids. Grossly, the lesion was yellow and distinct from other carcinoids in the gastric mucosa, mimicking gastric xanthelasma. Histologically, the tumor consisted of uniform clusters of polygonal cells with foamy cytoplasm. The formation of cell packets and abundant vascular stroma provided clues to its neuroendocrine nature. This was confirmed by immunoreactivity for chromogranin A and the demonstration of dense core granules ultrastructurally. PMID- 9341594 TI - Multiple myeloma arising from monoclonal gammopathy of undetermined significance in a patient with Gaucher's disease. AB - We report the case of a 64-year-old woman who, 12 years after receiving a diagnosis of Gaucher's disease with concurrent monoclonal gammopathy of undetermined significance, developed worsening thrombocytopenia and bone pain. Bone marrow biopsy at this time revealed 50% plasma cells with a serum monoclonal immunoglobulin A-lambda level of 3.2 g/L. Roentgenography revealed a lytic clavicular lesion, and a diagnosis of multiple myeloma was made. To our knowledge, this case is the first to document the evolution of a monoclonal gammopathy of undetermined significance to multiple myeloma in a patient with Gaucher's disease. The importance of investigating cytopenias and increased bone pain in such patients, perhaps by bone marrow biopsy, to rule out potentially malignant plasma cell dyscrasias is stressed. PMID- 9341596 TI - New American Association of Tissue Banks standards for semen banking. PMID- 9341595 TI - Peritoneal endometriosis, ovarian endometriosis, and adenomyotic nodules of the rectovaginal septum are three different entities. AB - OBJECTIVE: To review the histogenesis of peritoneal, ovarian, and rectovaginal endometriotic lesions. DESIGN: The comparison of morphologic, morphometric, and histochemical data observed in each type of lesion. SETTING: A university hospital department of gynecology. PATIENT(S): Patients complaining of infertility or pelvic pain with laparoscopically proved endometriosis. INTERVENTION(S): Laparoscopy was performed, and biopsy specimens from the endometriotic lesions were histologically studied. RESULT(S): Three types of endometriotic lesions must be considered: peritoneal, ovarian, and rectovaginal. Morphologic and morphometric data show similarities between eutopic endometrium and red peritoneal lesions, suggesting that these lesions are the first stage of early implantation of endometrial glands and stroma. After partial shedding, the red lesions regrow constantly. The shedding induces an inflammatory reaction, provoking scarification, and the lesions become black. The subsequent fibrosis leads to areas of white opacification that are inactive. The pathogenesis of ovarian endometriomas is a source of controversy. Although there seems to be a consensus concerning the invagination theory, there is still a contradiction between the implantation theory and the metaplasia theory. We recently showed that the mesothelium covering the ovary can invaginate into the ovarian cortex, pushing back the primordial follicles. The presence of mesothelial invagination in continuum with endometriotic tissue suggests that metaplastic histogenesis of ovarian endometriotic lesions occurs. Rectovaginal endometriotic nodules must be considered adenomyomas, consisting of smooth muscle with active glandular epithelium and scanty stroma. Immunocytochemical results show poor differentiation and hormonal independence of these lesions and indicate a close relation with their mesodermal mullerian origin. CONCLUSION(S): Peritoneal, ovarian and rectovaginal endometriotic lesions must be considered as three separate entities with different pathogeneses. PMID- 9341597 TI - Expectant management versus elective curettage for the treatment of spontaneous abortion. AB - OBJECTIVE: To determine whether the amount of intrauterine tissue was prognostic of the risk of complications associated with the management of nonviable pregnancies diagnosed in the first trimester before cervical dilatation (termed here impending abortion) with either expectant observation or elective curettage. DESIGN: Historic cohort study. SETTING: University Infertility Service. PATIENT(S): All women with nonviable pregnancies followed by the Division of Reproductive Endocrinology during a 5-year period. The patients were divided into those with significant intrauterine tissue (gestational sac > 10 mm) and those with minimal intrauterine tissue. INTERVENTION(S): Women either underwent elective curettage or were followed expectantly. MAIN OUTCOME MEASURE(S): Complication rates. RESULT(S): In 89 women with minimal tissue, no complications occurred regardless of treatment mode. In 63 women with significant intrauterine tissue, expectant management resulted in more complications (9/24) than elective curettage (1/39). In the expectant group, complications included missed abortion, septic abortion, and incomplete abortion requiring emergency curettage, with one patient requiring a transfusion. In the curettage group one uterine perforation occurred. CONCLUSION(S): In women with impending abortions and minimal intrauterine tissue, expectant treatment is safe after ectopic pregnancy has been excluded. In patients with significant intrauterine tissue, the risk of complications may be decreased by elective uterine curettage compared with expectant management. PMID- 9341598 TI - Follicular fluid vascular endothelial growth factor concentrations are elevated in women of advanced reproductive age undergoing ovulation induction. AB - OBJECTIVE(S): To determine whether follicular fluid (FF) from women of advanced reproductive age had a relative deficiency of the angiogenic cytokine vascular endothelial growth factor/vascular permeability factor. Furthermore, we sought to determine whether luteinized granulosa cells secrete vascular endothelial growth factor/vascular permeability factor in response to hypoxia. DESIGN: Retrospective cohort study. SETTING: University teaching hospital. PATIENTS: Women undergoing follicular aspiration after superovulation in preparation for IVF-ET. Women of advanced reproductive age consisted of 21 women > or = 38 years old (range, 38 to 46 years); 15 subjects < or = 30 years served as the control population. INTERVENTION(S): Granulosa cells and FF were collected by transvaginal aspiration 35 hours after hCG. Granulosa cells from two women were cultured for 24 and 48 hours in M199 + 10% fetal bovine serum in 1% O2-5% CO2-94% N2 (hypoxic) or 95% air-5% CO2 (normoxic) without or with 0.1 mol/L cobalt chloride. MAIN OUTCOME MEASURE(S): Pooled FF vascular endothelial growth factor/vascular permeability factor concentrations and media vascular endothelial growth factor/vascular permeability factor accumulation at 24 and 48 hours were determined. RESULT(S): Follicular fluid vascular endothelial growth factor/vascular permeability factor concentrations were higher in advanced reproductive age women compared with younger women (3,735 +/- 2,155 versus 2,205 +/- 952 pg/mL, mean +/- SD). Accumulation of vascular endothelial growth factor/vascular permeability factor at 24 and 48 hours was 391 +/- 54 and 744 +/- 2 pg/mL in media maintained in 5% CO2 and air. Cobalt chloride induced a marked increase in vascular endothelial growth factor/vascular permeability factor (2,008 +/- 52 pg/mL at 24 hours and 3,630 +/- 519 pg/mL at 48 hours). An intermediate but significant increase in vascular endothelial growth factor/vascular permeability factor (733 +/- 35 pg/mL at 24 hours and 2,675 +/- 864 pg/mL at 48 hours) was observed with 1% O2 compared with normoxic controls. CONCLUSION(S): After hMG and hCG administration the FF from women of advanced reproductive age showed increased vascular endothelial growth factor/vascular permeability factor concentrations compared with younger women. Increased vascular endothelial growth factor/vascular permeability factor concentrations could be consistent with a hypoxic environment within follicles of older women. PMID- 9341599 TI - Clinical relevance of the baboon as a model for the study of endometriosis. AB - OBJECTIVE: To review the value of the baboon as a model for the study of endometriosis. DATA IDENTIFICATION AND SELECTION: Studies performed at the Institute of Primate Research in Nairobi, Kenya (1990-1994), and published in peer-reviewed journals. RESULT(S): Spontaneous endometriosis was found in about 25% of the baboons, and its prevalence increased with the duration of captivity. The laparoscopic appearance, pelvic localization, and microscopic aspects of the disease were similar to endometriosis in women. Microscopic endometriosis in macroscopically normal peritoneum was rare. Sampson's hypothesis (i.e., retrograde menstruation causes endometriosis) was supported by the increased incidence of retrograde menstruation in baboons with spontaneous endometriosis, the observation that cervical occlusion could cause retrograde menstruation and endometriosis, and the finding that intrapelvic injection of menstrual endometrium caused experimental moderate to severe endometriosis similar to the spontaneous disease. During follow-up of more than 2 years, endometriosis in baboons appeared to be a progressive disease, with active remodeling between several types of lesions. Progression was stimulated by high-dose immunosuppression. Fertility was normal in baboons with minimal disease but was reduced in baboons with mild, moderate, or severe endometriosis, possibly related to an increased incidence and recurrence of the luteinized unruptured follicle syndrome. CONCLUSION(S): The baboon is a good model for the study of endometriosis. PMID- 9341600 TI - Factors associated with the quality before freezing and after thawing of sperm obtained by microsurgical epididymal aspiration. AB - OBJECTIVE: To identify whether the cause, site of ductal obstruction, and characteristics of fluid aspirates are associated with the cryosurvival and fertility after thawing of sperm obtained during reconstruction of the excurrent ducts with microsurgical epididymal sperm aspiration, vasal sperm aspiration, or both. DESIGN: Prospective study. SETTING: Andrology center at a tertiary care institution. PATIENT(S): Men undergoing reconstruction of the excurrent duct and sperm aspiration (n = 42) or microsurgical epididymal sperm aspiration (n = 11). INTERVENTION(S): Sperm were tested for an association with the cause and site of obstruction. Fertilization and pregnancy rates after sperm aspiration and intracytoplasmic sperm injection (ICSI) were evaluated for fresh and frozen aspirates. MAIN OUTCOME MEASURE(S): Motile sperm count and percentage motility after thawing. RESULT(S): The motile sperm count before freezing was significantly higher in the caput epididymis than in the corpus. The motile sperm count before freezing was related inversely to the distance from the caput where the sperm were aspirated. Sperm from clear and opaque fluid aspirates had better percent motility than those from cloudy and creamy fluid aspirates. High fertilization and pregnancy rates were achieved using both fresh and frozen epididymal sperm. CONCLUSION(S): None of the factors studied was associated with cryosurvival of aspirated epididymal or vasal spermatozoa. Because motility is low after thawing, these specimens are best used with ICSI. PMID- 9341602 TI - Epidemiologic and etiologic aspects of primary infertility in the Kashmir region of India. AB - OBJECTIVE: To assess the magnitude of primary infertility and to study its etiologic aspects in India. DESIGN: After proper randomization, 10,063 married couples were interviewed to ascertain the prevalence of primary infertility. A definitive protocol was followed to determine the etiology of primary infertility in 250 consecutive couples. SETTING: Tertiary care medical center in the Kashmir valley of India. PATIENT(S): Couples married for > or = 1 year; 250 consecutive couples attending an endocrine clinic for primary infertility. INTERVENTION(S): A logical investigative protocol was followed to identify the etiology of infertility. MAIN OUTCOME MEASURE(S): Magnitude of primary infertility in the community as well as the male, female, or combined etiology of infertility. RESULT(S): Fifteen percent of the couples interviewed had primary infertility, among whom 4.66% had unresolved infertility at the time of the survey. The etiology of infertility in 250 consecutive couples revealed a female factor in 57.6%, a male factor in 22.4%, combined factors in 5.2%, and an undetermined cause in 14.8%. CONCLUSION(S): Primary infertility is as common and distressing a problem in India as in other parts of the world. Semen abnormalities (22.4%), anovulation (17.2%), ovarian failure (8.8%), hyperprolactinemia (8.4%) and tubal disease (7.2%) are common causes of infertility. The pattern of infertility in India is the same as in other parts of the world, except that infertile couples report late for evaluation. PMID- 9341601 TI - Correlation between shrinkage of uterine leiomyoma treated with buserelin acetate and histopathologic findings of biopsy specimen before treatment. AB - OBJECTIVE: To analyze histopathologic features related to myoma volume reduction after treatment with a GnRH agonist, buserelin acetate, in needle biopsy specimens obtained before treatment. DESIGN: Prospective clinical study. SETTING: University teaching hospital. PATIENT(S): Twenty women with normal menstrual cycles and symptomatic uterine myomas. INTERVENTION(S): Buserelin acetate was administered intranasally (900 micrograms/d) for at least 16 weeks; transcervical needle biopsy of the uterine myoma was done before this treatment. MAIN OUTCOME MEASURE(S): The relation between histopathologic features (degree of cellularity, hyaline change, and collagen content) and the percent decrease in the volume of the largest myoma at 16 weeks measured by magnetic resonance imaging. RESULT(S): The correlation between the degree of hyaline change and the percent decrease in the myoma volume was significant, as was that with the proportion of collagenous tissue in the specimens. CONCLUSION(S): We predicted myoma volume reduction after treatment with a GnRH agonist by histologic analysis of the uterine leiomyoma before treatment. PMID- 9341603 TI - A comparison between two doses of flutamide (250 mg/d and 500 mg/d) in the treatment of hirsutism. AB - OBJECTIVE: To compare the effects of flutamide at 250 mg/d and 500 mg/d in the treatment of hirsutism. DESIGN: Randomized, prospective clinical study. PATIENT(S): Sixty-five patients with moderate to severe hirsutism. INTERVENTION(S): Group I (n = 35) patients were treated with flutamide, 250 mg/d, and group II (n = 30) patients were treated with flutamide, 500 mg/d, for 12 months. Baseline hormone levels, body mass index, and hirsutism scores were similar between the groups. Hirsutism score, hormone levels, and multiscreen blood chemistry were measured at 3-month intervals for 12 months. RESULT(S): The modified Ferriman-Gallwey scores for hirsutism decreased significantly at months 6 and 12 from a mean +/- SEM of 17.8 +/- 0.9 to 6.0 +/- 0.6 (P < 0.001) and 17.8 +/- 0.9 to 4.8 +/- 0.7 (P < 0.001) in group I and from 17.0 +/- 0.9 to 6.6 +/- 0.7 and 17.0 +/- 0.9 to 5.2 +/- 0.7 (P < 0.001) in group II, respectively. The reductions in hirsutism scores (mean +/- SEM) were similar in group I at 6 months (64.6% +/- 2.5%) and at 12 months (71.2% +/- 2.2%) and in group II at 6 months (62.1% +/- 3.0%) and at 12 months (70.3% +/- 3.0%). The percent reductions in hirsutiam scores at 6 and at 12 months were similar within group I (64.6% +/- 2.5% and 71.2% +/- 2.2%) and group II (62.1% +/- 3.0% and 70.3% +/- 3.0%), respectively. The decreases in hirsutism scores in the first 6 months were more significant than in the last 6 months of treatment in both groups. There were no significant differences in any of the hormone levels during therapy in either group. One patient in group II was excluded from the study because of liver dysfunction. CONCLUSION(S): This study shows that two different doses of flutamide (250 mg/d and 500 mg/d) are similarly effective in reducing hair growth. Flutamide at a dose of either 250 mg/d or 500 mg/d has no further effect after 6 months. We conclude that if flutamide is administered at a dose of 250 mg/d it may represent an effective and well tolerated treatment with reduced cost for the patient. PMID- 9341604 TI - Premature progesterone elevation spares blastulation but not pregnancy rates in in vitro fertilization with coculture. AB - OBJECTIVE: To clarify whether embryo development to the blastocyst stage may be affected by premature P elevation during controlled ovarian hyperstimulation (COH) for IVF-ET with embryo coculture. DESIGN: Retrospective study. SETTING: Tertiary care infertility center. PATIENT(S): One hundred thirty-one women undergoing 153 IVF-ET cycles with embryo coculture. INTERVENTION(S): Patients underwent COH with GnRH agonist and hMG. Embryos were cocultured up to the blastocyst stage. According to plasma P levels on the day of hCG, two groups were defined: low P (P < or = 0.9 ng/mL; conversion factor to SI unit, 3.180) and high P (P > 0.9 ng/mL). MAIN OUTCOME MEASURE(S): Blastulation (number of blastocysts/number of noncavitating embryos x 100) and pregnancy rates (PRs). RESULT(S): Blastulation rates were similar in the low and high P groups (51% and 48%, respectively). Moreover, patients included in the high P groups achieved significantly lower clinical and ongoing PRs (12% versus 29% and 7% versus 25%, respectively). CONCLUSION(S): The lack of difference in blastulation rates between the groups further supports the hypothesis that premature P elevation does not alter oocyte and embryo quality. Hence, the observed decrease in PRs is likely to reflect impaired endometrial receptivity in the high P group. PMID- 9341605 TI - Morphology and clinical outcomes of embryos after in vitro fertilization are superior to those after intracytoplasmic sperm injection. AB - OBJECTIVE: To compare embryos obtained after IVF and intracytoplasmic sperm injection (ICSI) regarding morphology and the likelihood of achieving clinical pregnancy. DESIGN: Case-control study. SETTING: An IVF unit controlling 1,310 cycles in 1996. PATIENT(S): Women having a total of 477 IVF and 475 ICSI consecutive cycles. INTERVENTION(S): Ovarian stimulation, IVF-ET, or ICSI-ET for all couples. MAIN OUTCOME MEASURE(S): Number of grade-A embryos transferred, preclinical pregnancy losses, and clinical pregnancy rates in IVF and ICSI cycles. RESULT(S): In comparison with the ICSI group, the IVF group showed significantly more grade-A embryos available for transfer (mean, 2 +/- 1.6 versus 1.8 +/- 1.5), significantly fewer preclinical pregnancy losses (1.6% versus 4%), and significantly higher clinical pregnancy rates (25% versus 19.1%). CONCLUSION(S): Embryos obtained after IVF are superior to those obtained after ICSI in relation to embryo morphology and the likelihood of achieving clinical pregnancy. PMID- 9341606 TI - Lysis of intrauterine adhesions using gynecoradiologic techniques. AB - OBJECTIVE: To present further experience with in-office lysis of intrauterine adhesions under fluoroscopic control using a specially designed catheter. DESIGN: Prospective study. SETTING: Medical school-affiliated infertility center. PATIENT(S): Seventeen infertile patients undergoing routine gynecoradiologic investigation as part of an initial infertility workup. INTERVENTION(S): The initial hysterosalpinography was performed with a commercially available uterine catheter that seals off the uterine cavity before injection of contrast. If intrauterine adhesions were diagnosed, an immediate attempt at lysis was made using the catheter's balloon tip or hysteroscopic scissors, which were inserted through the main port of the catheter. The procedures were carried out using a paracervical block or IV analgesia. MAIN OUTCOME MEASURE(S): Normal uterine cavity after lysis of intrauterine adhesions. RESULT(S): Seventeen patients underwent lysis of intrauterine adhesions. In 13 patients (9 mild, 3 moderate, and 1 severe), the adhesions were lysed successfully (81.2%). Among those, nine procedures were performed with the balloon and four with scissors. In 4 cases (2 moderate and 2 severe), lysis of adhesions was only partially successful. These procedures had to be abandoned prematurely because of patient discomfort before attempting the use of scissors (n = 1), extravasation of dye into the myometrium making visualization difficult (n = 1), and thick, fibrotic adhesions that were resistant to scissors (n = 2). CONCLUSION(S): In-office lysis of intrauterine adhesions under gynecoradiologic control can be carried out safely in the majority of patients, using minimally invasive techniques. The potential cost savings in comparison with endoscopic procedures, which require utilization of expensive operating room time, are especially relevant in today's cost-conscious managed care environment. Only failures of in-office procedures would reach the operating room under the algorithm proposed here. PMID- 9341607 TI - Description of a laparoscopic technique for treating patients with ovarian remnant syndrome. AB - OBJECTIVE: A retroperitoneal approach for laparoscopic treatment of ovarian remnant syndrome was developed. DESIGN: Clinical study. SETTING: Department of Gynecology, Friedrich-Schiller-University Jena. PATIENT(S): During a 29-month period, seven consecutive patients with ovarian remnant syndrome were treated by laparoscopy. Patients were not preselected and preoperative, intraoperative, and postoperative data were registered prospectively. INTERVENTION(S): For removal of remnant ovaries we used a laparoscopic retroperitoneal approach that included complete dissection of the pelvic course of the ureter and coagulation and dissection of the infundibulopelvic ligament and of the uterine vessels. RESULT(S): In the first patient's case, the right ureter was injured during dissection, which was initiated too far distally between ovary and external iliac vessels. Thereafter, we changed our technique to start the dissection of the ureter at the pelvic brim. No subsequent patient had an intraoperative or postoperative complication. All patients reported fewer preoperative complaints and were free of recurrence by sonographic examination. CONCLUSION(S): Using a retroperitoneal approach laparoscopic resection of a remnant ovary may be a safe and effective technique. PMID- 9341608 TI - Effect of human hydrosalpinx fluid on murine embryo development and implantation. AB - OBJECTIVE: To determine the effect of hydrosalpinx fluid-containing medium on murine embryo development and implantation. DESIGN: The development of one-, two , and four-cell mouse embryos in medium containing 5%, 10%, and 20% of human hydrosalpinx fluid was observed. Implantation rates of mouse embryos transferred into the uterine horn with hydrosalpinx fluid-containing media were determined. SETTING: Private hospital-based fertility center and IVF program. MAIN OUTCOME MEASURE(S): Percentage of embryos continuing cell division and implantation rates after ET. RESULT(S): Hydrosalpinx fluid in culture medium affected embryo development in a dose-dependent fashion. The injection of hydrosalpinx fluid containing medium into the uterine horn did not affect embryo implantation. CONCLUSION(S): Hydrosalpinx fluid negatively affects murine embryo development, but its presence in the uterine horn at ET did not affect implantation. PMID- 9341609 TI - The influence of clinical and subclinical varicocele on testicular volume. AB - OBJECTIVE: To examine the possible loss of testicular volume in infertile men with clinical and subclinical varicocele by using ultrasound (US)-derived measurements of testicular volume. DESIGN: Retrospective review of clinical and scrotal US reports. SETTING: University infertility clinic. PATIENT(S): Infertile men (n = 404) presenting for evaluation from 1992 to 1996. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Presence of clinical or subclinical varicocele, US derived measurements of testicular volume. RESULT(S): In men with clinical left or subclinical left varicocele, left testicular volume was significantly less than right testicular volume (12.9 versus 14.1 and 13.2 versus 14.7 mL, respectively). This finding was not observed in men with bilateral clinical or bilateral subclinical varicoceles or in men without varicocele. CONCLUSION(S): Our data confirm previous reports showing that a clinical left varicocele can negatively impact on left testicular volume and for the first time show that a subclinical varicocele is also associated with decreased left testicular volume. PMID- 9341610 TI - Male immunologic infertility: sperm performance on in vitro fertilization. AB - OBJECTIVE: To analyze sperm performance in a group of patients with male immunologic infertility treated with IVF-ET. DESIGN: Retrospective clinical study. SETTING: Patients attending a private IVF clinic. PATIENT(S): The study group comprised seven men with significant levels of surface-bound antisperm antibodies treated in nine IVF cycles. The control group comprised nine couples with female tubal infertility and no indication of male factor infertility treated on the same cycle. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Fertilization rate, early embryonic development, implantation, and clinical pregnancy rate (PR). RESULT(S): Forty-six (44.2%) of 104 inseminated oocytes were fertilized in the study group compared with 65 (84.4%) of 77 in the control group, which was a significant difference. Surface-bound antisperm antibodies significantly inhibited early embryonic cleavage in the study group (13 [28.3%] of 46 embryos with at least 3 blastomeres) compared with the control group (41 [63.1%] of 65 embryos, with at least 3 blastomeres). The percentage of good quality embryos (grades 1 and 2) was similar in the study and control groups (71.7% and 78.5%, respectively). The percentage of poor-quality embryos (grade 4 and two pronuclei) was higher in the study group compared with the control group (13.9% versus 9.2%, respectively); however, the difference was not significant. The implantation rate and clinical PR were lower in the study group (3% and 11%, respectively) compared with the control group (9.5% and 44%, respectively), but the difference was not statistically significant. CONCLUSION(S): The fertilization rate and early embryonic cleavage of human embryos was found to be reduced significantly in patients with high levels of surface-bound antisperm antibodies. Moreover, embryonic quality and the PR may be compromised by the presence of significant levels of surface-bound antisperm antibodies. PMID- 9341611 TI - Mechanical isolation and in vitro growth of preantral and small antral human follicles. AB - OBJECTIVE: To develop a procedure for isolating small human follicles and to determine their growth requirements. DESIGN: Preantral and early antral follicles were isolated manually, allocated randomly to experimental groups, and cultured for a few weeks. SETTING: Patients giving informed consent in hospitals. PATIENT(S): Women undergoing laparotomy or oophorectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Follicular size, E2, histology. RESULT(S): Human FSH (at a dose of 1.5 U/mL) induced antral growth of follicles, and the addition of human LH (2.5 ng/mL) to human FSH stimulated growth and antral development. Histologic studies showed that most of the early antral follicles did not contain an oocyte and already had begun to undergo atresia before culturing. Levels of E2 increased in the incubation medium as the follicles increased in size, but those levels were significantly greater when the follicles contained oocytes. CONCLUSION(S): It is possible to grow small human follicles after they have been isolated manually. To develop successfully, they require a low concentration of human LH in addition to human FSH. The rate of atresia between the preantral and early antral stages in vivo is very high; therefore, it is worthwhile to develop techniques for isolating and culturing the follicles before the antral stages. PMID- 9341612 TI - Expression of glutathione peroxidases in the adult male rat reproductive tract. AB - OBJECTIVE: To evaluate the messenger RNA (mRNA) expression of three glutathione peroxidase isoforms in the male reproductive tract and to further characterize testicular glutathione peroxidase expression. DESIGN: Analysis of glutathione peroxidase levels in untreated animals. INTERVENTION(S): 32P-labeled DNA probes were derived from known complementary DNA (cDNA) sequences for classic cellular glutathione peroxidase, phospholipid hydroperoxide glutathione peroxidase, and secretory epididymal glutathione peroxidase, and used to evaluate mRNA levels in each tissue by Northern blot hybridization. MAIN OUTCOME MEASURE(S): Glutathione peroxidase mRNA concentrations. RESULT(S): A 0.8-kb transcript was identified in liver, testis, prostate, seminal vesicle, vas deferens, and epididymis using the cDNA probe for classic cellular glutathione peroxidase. Using the probe for phospholipid hydroperoxide glutathione peroxidase, a 0.9-kb transcript was identified in the epididymis, vas deferens, prostate, seminal vesicle, and liver. In the testis, the phospholipid hydroperoxide glutathione peroxidase transcript was highly abundant and longer, measuring 1.1 kb. The phospholipid hydroperoxide glutathione peroxidase mRNA transcript was expressed in 40-, 60-, and 90-day-old rat testes, but was undetectable in testes of 10- and 20-day-old rats. Epididymal glutathione peroxidase was detected as a single 1.9-kb transcript in the caput epididymis only. CONCLUSION(S): Male rat reproductive tissues express at least three different isozymes of glutathione peroxidase. Phospholipid hydroperoxide glutathione peroxidase and classic cellular glutathione peroxidase are primarily found in testis, whereas epididymal glutathione peroxidase is expressed in the epididymis. PMID- 9341613 TI - Prolactin gene expression and prolactin protein in premenopausal and postmenopausal human ovaries. AB - OBJECTIVE: To investigate intraovarian prolactin and prolactin-receptor gene expression and to assess local prolactin synthesis with emphasis on possible differences between premenopausal and postmenopausal status. DESIGN: The RNA extracted from human premenopausal and postmenopausal tissues was subjected to reverse transcription and polymerase chain reaction by using prolactin-specific intron- and exon-spanning primers. Prolactin-receptor expression was investigated accordingly. The amplified complementary DNA fragments were analyzed by gel electrophoresis and restriction enzyme mapping. Local prolactin hormone synthesis was verified by a time-resolved immunofluorometric assay based on our monoclonal antibodies. RESULT(S): Prolactin and prolactin-receptor gene expression was observed in all analyzed human ovaries (n = 18). Several other human tissue specimens, such as lung and kidney, served as negative control tissues. Significantly elevated concentrations of prolactin were detected in cytosolic extracts of premenopausal (n = 6; mean +/- SD; 20.6 +/- 3.3 ng/g tissue wet weight) versus postmenopausal (n = 6; 3.6 +/- 3.0 ng/g tissue wet weight) ovaries. CONCLUSION(S): The human ovary not only serves as a target for endocrine prolactin action but also as a site of local prolactin hormone production. In agreement with previous reports on extrapituitary sources of prolactin, we consider prolactin as a hormone as well as an autocrine or paracrine growth or regulatory factor. Significantly increased concentrations of prolactin in premenopausal ovarian tissue verifies its role in human reproduction. PMID- 9341614 TI - Allelic polymorphism and chromosomal localization of the human oviductin gene (MUC9). AB - OBJECTIVE: To determine if the gene coding for human oviductin (the estrogen dependent, oviduct-specific glycoprotein with an affinity for the zona pellucida) shows length polymorphism in the region of tandem repeats. To determine the frequencies of the length alleles in health and disease. DESIGN: Descriptive fundamental and clinical studies. SETTING: Fertility clinic and research center, university teaching hospital. PATIENT(S): Fertile women, women with a history of ectopic pregnancy or tubal disease, and women with stage I or II endometriosis. INTERVENTION(S): Blood samples were drawn for DNA analysis. MAIN OUTCOME MEASURE(S): Length and sequence of the region of tandem repeats. RESULT(S): Four different length alleles of the human oviductin gene were identified. Their relative frequencies in pathologic cases were not statistically significant compared with those found in normal fertile women. The human genome contains a single copy of the oviductin gene located on chromosome 1p13. CONCLUSION(S): The human oviductin gene codes for a glycoprotein that shares the characteristics of epithelial mucins. Because eight epithelial mucin genes have been identified so far, we therefore propose to name this gene MUC9. The biologic function of the protein is likely to include protection of the early embryo and of the fallopian tube itself. PMID- 9341615 TI - Measurement of vitronectin content of human spermatozoa and vitronectin concentration within seminal fluid. AB - OBJECTIVE: Vitronectin previously has been extracted from human spermatozoa and messenger RNA (mRNA) encoding vitronectin localized by reverse transcriptase in situ polymerase chain reaction (PCR) to spermatocytes of human testis. In the present experiments, we have established ranges for the content of vitronectin in living human spermatozoa and vitronectin concentration within seminal fluid of human ejaculates. DESIGN: Seminal fluid was obtained by centrifugation and motile sperm selected by swim-up from men with normal and abnormal ejaculates, according to World Health Organization criteria, for vitronectin determinations. SETTING: Academic research environment. MAIN OUTCOME MEASURE(S): Seminal fluid vitronectin concentrations were measured by ELISA and sperm vitronectin content by polyacrylamide gel electrophoresis and semiquantitative Western blots. RESULT(S): Vitronectin seminal fluid concentration was 1.35 +/- 1.0 mg/mL (mean +/- SD) for normospermic samples (n = 26) and 0.78 +/- 0.4 mg/mL for azoospermic specimens (n = 6). Vitronectin sperm content ranged from 1 to 15 ng/10(6) motile cells (n = 20). Both high- and low-molecular-weight material was observed. Sperm content of vitronectin did not vary with sperm morphology. CONCLUSION(S): These results suggest that spermatozoa represent a major source of seminal fluid vitronectin, but that a secondary source exists, perhaps through transudation from serum. PMID- 9341616 TI - Failed fertilization after intracytoplasmic sperm injection: the extent of paternal and maternal chromatin decondensation. AB - OBJECTIVE: To determine the extent of paternal and maternal chromatin decondensation in unfertilized eggs after intracytoplasmic sperm injection (ICSI). DESIGN: Eggs that failed to show two pronuclei (2-PN) 48 hours after ICSI were studied at two different time intervals: at ICSI program inception (group A) and after 8 months (group B). PATIENT(S): Forty-nine patients undergoing IVF cycles. MAIN OUTCOME MEASURE(S): The unfertilized eggs were studied by chromatin staining. RESULT(S): The average fertilization rate from all ICSI cycles in these two groups was 45%. The fertilization rates in groups A and B were 35% and 59%, respectively. In group A, 65% of the unfertilized eggs were characterized by condensed sperm chromatin with 11% showing partial decondensation. In group B, only 28% of the unfertilized eggs demonstrated condensed sperm chromatin, whereas 45% were partially decondensed. In these two groups, no sperm chromatin was detected in 24% of the unfertilized eggs. The maternal chromatin remained at metaphase II in 84% of all unfertilized eggs analyzed. CONCLUSION(S): These observations suggest that the technical problem of deposition of the sperm inside the egg is not the major cause of failure of fertilization rates in ICSI cycles. Rather, it is likely to be the failure to complete both the maternal and paternal chromatin transitions that occur with normal fertilization. PMID- 9341617 TI - Pregnancy after intracytoplasmic sperm injection with sperm from a man with a 47,XXY Klinefelter's karyotype. AB - OBJECTIVE: To report the initiation of a pregnancy that was achieved by intracytoplasmic sperm injection (ICSI) with sperm from a patient with Klinefelter's syndrome. DESIGN: Case report. SETTING: University women's hospital IVF center. PATIENT(S): A couple with primary infertility and nonmosaic 47,XXY karyotype of the male partner. INTERVENTION(S): Intracytoplasmic sperm injection after ovarian stimulation and transvaginal ultrasound-guided oocyte pick-up with sperm from a hypergonadotropic man with a nonmosaic 47,XXY karyotype. MAIN OUTCOME MEASURE(S): Clinical pregnancy. RESULT(S): Despite a 47,XXY karyotype in all 50 analyzed lymphocyte metaphases, the sperm of the patient led to a clinical pregnancy with the first attempt of ICSI and intrauterine transfer of three embryos. The pregnancy stopped developing in the ninth week. Cytogenetic investigation of the abortion material revealed a numerical normal 46,XXY karyotype. CONCLUSION(S): Sperm from a patient with hypergonadotropic nonmosaic Klinefelter's syndrome, when used for ICSI, can lead to a pregnancy. PMID- 9341618 TI - Removal of surgical barriers of expanded polytetrafluorethylene at second-look laparoscopy was not associated with adhesion formation. AB - OBJECTIVE: To evaluate whether the surgical trauma required for the laparoscopic removal of a polytetrafluoroethylene (PTFE) surgical barrier would cause postoperative adhesions. DESIGN: Two case reports. SETTING: Tertiary academic medical center. PATIENT(S): Two women who had undergone myomectomy with placement of a PTFE surgical barrier and who were free of adhesions with the barrier in place. INTERVENTION(S): Removal of the PTFE barrier by laparoscopy 11 days after myomectomy. MAIN OUTCOME MEASURE(S): Adhesions at the site of removal of the PTFE barrier at the time of incidental laparoscopy several years later. RESULT(S): Adhesions were not present at the site of PTFE barrier removal. CONCLUSION(S): The surgical trauma required to remove PTFE barriers at early second-look laparoscopy was not associated with postoperative adhesions. PMID- 9341619 TI - Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. AB - OBJECTIVE: To describe the first birth achieved after intracytoplasmic sperm injection (ICSI) of cryopreserved human oocytes. DESIGN: Case report. SETTING: University of Bologna Hospital, Department of Obstetrics and Gynecology, Reproductive Endocrinology Unit, IVF and Infertility Center. PATIENT(S): One patient undergoing IVF. INTERVENTION(S): Transvaginal ultrasound-guided oocyte retrieval followed by oocyte freezing. Artificial preparation of the endometrium with E2 and P, oocyte thawing, and ICSI. RESULT(S): Four of 12 cryopreserved oocytes survived; using ICSI, 2 underwent normal fertilization but only 1 cleaved. One good-quality 4-cell embryo was transferred. A single gestation was confirmed by ultrasound at the 7th week. Amniocentesis was performed at the 16th week and demonstrated a normal female karyotype of 46,XX. After a normal pregnancy, a healthy female infant was born at the 38th week of gestation. CONCLUSION(S): The combination of ICSI and oocyte cryopreservation is a new tool in assisted reproductive technology. PMID- 9341620 TI - Medical treatment of ectopic pregnancy with methotrexate. AB - OBJECTIVE: To review our experience with low-dose IM methotrexate for the medical management of ectopic pregnancy (EP). DESIGN: Retrospective chart review. SETTING: Magee-Womens Hospital, Pittsburgh, Pennsylvania. PATIENT(S): The first 50 women treated by the resident service in whom EP was diagnosed and treated with methotrexate. INTERVENTION(S): Intramuscular methotrexate, 50 mg/m2. Serum beta-hCG was evaluated 4 and 7 days after treatment and then weekly thereafter. The dose was repeated if the beta-hCG level did not drop > or = 15% between days 4 and 7 or if a plateau or rise was noted during weekly follow-up evaluation. Surgery was performed if significant abdominal pain occurred in the presence of hemodynamic instability or signs of peritoneal irritation on physical examination. MAIN OUTCOME MEASURE(S): Resolution of the EP without surgical intervention. RESULT(S): Two patients were lost to follow-up and one was treated without a certain diagnosis of EP. Forty-three of the remaining 47 women (91.5%; 95% confidence interval, 83.5%, 99.5%) were treated successfully with methotrexate. Of these, 36 women were treated with a single dose, and 7 required a second dose. Four women were treated surgically after medical management failed. The time from initiation of treatment to cure in women who were treated successfully was 25 +/- 15 days (mean +/- SD). Thirteen patients (27.7%) made additional visits to the emergency department because of increased abdominal pain. CONCLUSION(S): As medical therapy for EP becomes common practice, familiarity with its side effects may lead to greater success rates. The decision to abandon medical treatment and proceed with surgery should be based on defined guidelines, such as the development of peritoneal signs, decreasing hemoglobin levels, or hemodynamic instability. PMID- 9341621 TI - Ultrasonographic endometrial changes after intrauterine insemination: a comparison of two catheters. AB - OBJECTIVE: To determine and compare the transvaginal ultrasonographic (US) endometrial changes immediately after IUI using the Edwards Wallace (H.G. Wallace, Limited, Colchester, Essex, UK) and Tom-Cat (Sherwood Medical, St. Louis, MO) catheters. DESIGN: Prospective study. SETTING: IVF unit. PATIENT(S) AND INTERVENTION(S): Eighty-two infertile patients underwent 112 cycles of ovulation induction with IUI. Either the Edwards Wallace catheter (group 1, n = 32) or the Tom-Cat catheter (group 2, n = 80) was used for sperm insemination. The presence of an endometrial three-layer pattern before IUI was a prerequisite for inclusion in the study. After each IUI, the endometrium was reassessed by transvaginal US. MAIN OUTCOME MEASURE(S): Ultrasonographic endometrial changes, clinical pregnancy rates (PRs), complications, and patients' complaints were compared between the two groups. RESULT(S): Total destruction of the endometrial three-layer pattern was observed in 12.5% of the cycles in group 1, compared with 50% of the cycles in group 2. Clinical pregnancies occurred in 14 (12.5%) of the 112 IUI cycles. A higher PR was achieved when the endometrial three-layer pattern was preserved after IUI. The patients in group 2 had more complaints of bleeding and pain during the procedure. CONCLUSION(S): Ultrasonographic changes after IUI suggest that the Edwards Wallace catheter is significantly less traumatic to the endometrium than the Tom-Cat catheter. Although both catheters yielded the same overall PR, there was a trend indicating that sparing the endometrial three-layer pattern from damage increases the chance of conception. PMID- 9341622 TI - Improving the intracytoplasmic sperm injection technique by transmembrane electric potential monitoring. AB - OBJECTIVE: To evaluate electrophysiologic techniques for transmembrane potential measurement during intracytoplasmic sperm injection (ICSI). DESIGN: Mature mouse oocytes were subjected to intracellular measurements of membrane potential using conventional techniques and modifications of the techniques for use with sham ICSI. During the procedure, the actual penetration of the membrane was determined subjectively or objectively with or without monitoring membrane potential. SETTING: Academic medical center. ANIMAL(S): Mouse. INTERVENTION(S): Measurement of oocyte membrane potential (Sham ICSI). MAIN OUTCOME MEASURE(S): Detectability of transmembrane potential using different electrodes and comparison of subjective and objective determination of membrane penetration. RESULT(S): Measurement of the membrane potential with the same glass pipettes used in ICSI means compromise between signal amplitude and compatibility with the conventional ICSI setup. Signal quality is related inversely to the diameter of the injection pipette, and its amplitude decreases as the concentrated electrode filling solution is replaced by physiologic solutions used in ICSI. When successful membrane penetration is suspected by visualization, measuring the potential at the tip of the injection pipette often proves otherwise. Conversely, when membrane penetration is confirmed by detection of transmembrane potential, the procedure may appear subjectively unsuccessful. CONCLUSION(S): Monitoring of transmembrane potential can be done successfully in conjunction with standard ICSI and has several potential applications. PMID- 9341623 TI - Identification of clear vesicular lesions of atypical endometriosis: a new technique. AB - OBJECTIVE: To describe a new technique for improvement of visualization and identification of clear vesicles of endometriosis. DESIGN: A new descriptive technique. SETTING: Tertiary care center. PATIENT(S): Adolescent and adult women with pelvic pain undergoing laparoscopic evaluation. INTERVENTION(S): The pelvis is filled with irrigation fluid, and then the laparoscope is submerged, with evaluation of the pelvic peritoneum through the liquid medium. MAIN OUTCOME MEASURE(S): Identification of clear lesions of endometriosis. RESULT(S): Laparoscopic visualization of clear lesions of atypical endometriosis can be facilitated with the use of liquid introduced into the pelvis in order to identify and appreciate the three-dimensional configuration of lesions and to exclude light reflection as a possible etiology. CONCLUSION(S): This method is suggested for identification of subtle lesions of endometriosis, which will aid in the establishment of the definitive diagnosis for adolescents and adults with chronic pelvic pain and will decrease the number of cases of unrecognized atypical endometriosis. PMID- 9341624 TI - A new method for freezing testicular biopsy sperm: three pregnancies with sperm extracted from cryopreserved sections of seminiferous tubule. AB - OBJECTIVE: To determine whether spermatozoa, located in the seminiferous tubules obtained by needle puncture testicular biopsy, could be cryopreserved successfully within the tubules and subsequently be used for in oocyte fertilization via intracytoplasmic sperm injection (ICSI) after the spermatozoa were removed from the thawed tubules. DESIGN: Clinical case series. SETTING: Private IVF unit. PATIENT(S): Six azoospermic patients (four obstructive, two maturation arrest). MAIN OUTCOME MEASURE(S): Survival rate of thawed spermatozoa, fertilization rate of oocytes after ICSI with spermatozoa extracted from thawed tubules and pregnancies. RESULT(S): All six patients had adequate motile spermatozoa extracted from the thawed tubule sections, and all patients achieved fertilization via ICSI with their partner's eggs. The fertilization rate was 46%, compared with 56% obtained in other previous patient cycles using fresh testicular spermatozoa. Three pregnancies resulted from five ETs. CONCLUSION(S): Cryopreservation and subsequent thawing of seminiferous tubules proved to be a simple and successful method for storage of testicular spermatozoa, reducing the need for repetitive testicular biopsies and increasing the likelihood of sperm availability on the day of oocyte retrieval. PMID- 9341625 TI - Causality and association--more than just different shades of gray. PMID- 9341626 TI - Small "time to event" studies--value? PMID- 9341627 TI - Number of embryos transferred and age of mother. PMID- 9341628 TI - ART and high risk patients! PMID- 9341629 TI - Easing the pain of death. PMID- 9341630 TI - Massive hemoptysis in a woman with seizures. AB - A 53-year-old woman presented with a productive cough, fever, chills, and night sweats of one month's duration. She reported having had lightly blood-streaked sputum initially but then experiencing massive hemoptysis (> 200 mL/2 hr). Since the onset of symptoms, she had had malaise, body aches, and a 27-lb weight loss. For the last two weeks, she had also had increasing shortness of breath and pleuritic chest pain. PMID- 9341631 TI - A pulse of 27 in a man with chronic heart disease. PMID- 9341632 TI - Long QT interval. PMID- 9341633 TI - Long QT interval. PMID- 9341634 TI - Estrogen replacement. PMID- 9341635 TI - Maimonides and mind-body medicine. PMID- 9341636 TI - Breast cancer: the high-risk mutations. AB - Mutations in the two known BRCA genes account for only 5% to 10% of breast cancer. The genes may play a role in genomic stability; their role in sporadic cancer remains unknown. Hence, genetic testing has created clinical dilemmas. Some pertain to women whose family history suggests inherited risk. Others, including the false reassurance that may be conferred by a negative finding, apply throughout the general population. PMID- 9341638 TI - Peripheral neuropathy in the nondiabetic patient. AB - The differential diagnosis includes an array of acquired and hereditary disorders. The history often offers the richest source of information, which can then direct the physical examination and electrodiagnostic testing. Further studies may include nerve biopsy and, most recently, blood testing for genetic disorders. Some neuropathies may respond to treatment; indeed, the response to treatment may be diagnostic. PMID- 9341637 TI - Acute exacerbations of chronic bronchitis. AB - Not every patient with bronchitis needs to be treated with an antibiotic. When treatment is indicated, however, the regimen should be selected carefully. A simple four-part disease classification scheme serves as a practical aid for initial assessment of the patient and as a guideline for choosing therapy. PMID- 9341639 TI - Unstable angina: integrating new treatments. AB - Although aspirin plus heparin remains the gold standard, clinicians will soon be able to select, from an array of antithrombotic strategies, the agents that best match the patient's hemodynamic status. Revascularization is not routinely indicated; it should be reserved for high-risk patients in whom medical therapy has failed, who have had a myocardial infarction, or who have other compromising factors. PMID- 9341640 TI - Prostate disease in older men: 2. Cancer. AB - Proof is lacking that early detection and treatment reduce mortality from prostate cancer. Indeed, many patients die with the tumor, not from it. In the absence of better data, the clinician must, when making treatment recommendations, weigh the biology of the disease, the potential morbidity of treatment, other factors that affect the patient's longevity, and the patient's attitude toward risk. PMID- 9341641 TI - Causes and treatment of loose bodies in the knee. PMID- 9341642 TI - Appropriateness guidelines to balance quality and cost. PMID- 9341643 TI - Pernicious vomiting in two pregnant women. PMID- 9341673 TI - The gene coding for the DOPA dioxygenase involved in betalain biosynthesis in Amanita muscaria and its regulation. AB - Genomic and cDNA clones derived from the gene (dodA) coding for DOPA dioxygenase, a key enzyme in the betalain pathway, were obtained from the basidiomycete Amanita muscaria. A cDNA library was established in the phage lambda ZapII and dodA clones were isolated using polyclonal antibodies raised against the purified enzyme. Their identity was confirmed by comparison of the deduced amino acid sequence with the sequence of several tryptic peptide fragments of DOPA dioxygenase. The gene coded for a 228-amino acid protein that showed no homology to published sequences. The coding region was interrupted by five short introns. Regulation was shown to occur at the transcriptional level; the mRNA accumulated to high levels only in the coloured cap tissue. dodA was found to be a single copy gene in A. muscaria. To our knowledge, this is the first gene from the betalain pathway to be cloned. It encodes a type of aromatic ring-cleaving dioxygenase that has not been previously described. PMID- 9341674 TI - Mutations at the human minisatellite MS32 integrated in yeast occur with high frequency in meiosis and involve complex recombination events. AB - Minisatellites are composed of tandem repetitive DNA sequences and are present at many positions in the human genome. They frequently mutate to new length alleles in the germline, by complex and incompletely understood recombination mechanisms which may operate during meiosis. In several minisatellites the mutation events are restricted to one end of the repeat array, indicating a possible association with elements that act in cis. Mutant alleles do not show exchange of flanking regions. To construct a model system suitable for further investigations of the mutation process, we have integrated the human minisatellite MS32, flanked by synthetic markers, in the vicinity of a meiotic recombination hot spot upstream of the LEU2 locus in the yeast Saccharomyces cerevisiae. Here we provide direct evidence for a meiotic origin of MS32 mutations. Mutation events were polarised towards both ends of the minisatellite and varied from simple duplications and deletions to complex intra- and interallelic events. Interallelic events were frequently accompanied by exchange of regions flanking the minisatellite. The results also support the notion that cis-acting elements are involved in the mutational process. The fact that MS32 mutant structures are similar in yeast and human shows that meiotic recombination plays a crucial role in both organisms and emphasises the usefulness of yeast strains harbouring minisatellites as a model system for the study of minisatellite mutation. PMID- 9341675 TI - Trichoderma reesei sequences that bind to the nuclear matrix enhance transformation frequency. AB - Three DNA fragments, trs1, 2 and 3, were isolated from the Trichoderma reesei genome on the basis of their ability to promote autonomous replication of plasmids in Saccharomyces cerevisiae. Each trs element bound specifically to the isolated T. reesei nuclear matrix in vitro, and two of them bound in vivo, indicating that they are matrix attachment regions (MARs). A similar sequence previously isolated from Aspergillus nidulans (ans1) was also shown to bind specifically to the T. reesei nuclear matrix in vitro. The T. reesei MARs are AT rich sequences containing 70%, 86% and 73% A + T over 2.9, 0.8 and 3.7 kb, respectively for trs1, 2 and 3. They exhibited no significant sequence homology, but were shown to contain a number of sequence motifs that occur frequently in many MARs identified in other eukaryotes. However, these motifs occurred as frequently in the trs elements as in randomly generated sequences with the same A + T content. trs1 and 3 were shown to be present as single copies in the T. reesei genome. The presence of the trs elements in transforming plasmids enhanced the frequency of integrative transformation of T. reesei up to five fold over plasmids without a trs. No evidence was obtained to suggest that the trs elements promoted efficient replication of plasmids in T. reseei. A mechanism for the enhancement of transformation frequency by the trs elements is proposed. PMID- 9341676 TI - Restriction enzyme-mediated DNA integration in Coprinus cinereus. AB - Restriction enzyme-mediated DNA integration (REMI) has recently received attention as a new technique for the generation of mutants by transformation in fungi. Here we analyse this method in the basidiomycete Coprinus cinereus using the homologous pabI gene as a selectable marker and the restriction enzymes BamHI, EcoRI and PstI. Addition of restriction enzymes to transformation mixtures results in an earlier appearance of transformants and influences transformation rates in an enzyme- and concentration-dependent manner. Low concentrations of restriction enzyme result in increased numbers of transformation rates decrease with higher enzyme concentrations. If protoplasts are made from cells stored in the cold, the transformation rates drop drastically even in the presence of low amounts of enzyme. In several transformants, plasmid integration directly correlated with the action of restriction enzyme at random chromosomal restriction sites. In some cases, restriction enzymes appear to reduce the number of integration events per transformant. Simultaneously, mutation rates can be enhanced due to the presence of restriction enzymes. Although restriction enzymes clearly promote plasmid integration into the host genome they also have cytotoxic and possibly mutagenic effects that result from processes other than plasmid integration. In consequence, for any given enzyme used in REMI mutagenesis, the enzyme concentration that gives the highest number of transformants must be defined experimentally. Such optimal transformation conditions should give the highest probability of obtaining mutations caused by a single restriction enzyme mediated integration of the selection marker. PMID- 9341677 TI - Differential display analysis of RNA accumulation in arbuscular mycorrhiza of pea and isolation of a novel symbiosis-regulated plant gene. AB - Differential RNA display was used to analyze gene expression during the early steps of mycorrhiza development on Pisum sativum following inoculation with Glomus mosseae. Seven out of 118 differentially displayed cDNA fragments were subcloned and sequenced. One fragment corresponded to part of the fungal 25S ribosomal RNA gene and a second one showed similarity to a human Alu element. The others were derived from plant genes of unknown function. One of the fragments was used for the isolation of a full-length cDNA clone. It corresponded to a single-copy gene (psam1) which is induced during early symbiotic interactions, and codes for a putative transmembrane protein. Northern and RNA dot blot analyses revealed enhanced accumulation of psam1 RNA after inoculation with G. mosseae of wild-type pea and an isogenic mutant deficient for nodule development (Nod-, Myc+). PMID- 9341678 TI - Overexpression of the protein kinase Pak1 suppresses yeast DNA polymerase mutations. AB - This article presents the identification and characterization of the PAK1 gene of Saccharomyces cerevisiae, and the biochemical characterization of the protein kinase activity that it encodes. Overexpression of the PAK1 gene product suppresses temperature-sensitive mutations of the poll (cdc 17) gene, which encodes DNA polymerase alpha. Overexpression and suppression can be achieved either by expressing PAK1 from a high-copy-number plasmid, or by GAL1-induced transcription of PAK1. Gene disruption of PAK1 indicates that it is not an essential gene. The PAK1 gene encodes a protein with a kinase consensus domain. By deletion analysis and site-directed mutagenesis, we demonstrate that the complete and active kinase consensus domain is required for suppression. A glutathione-S-transferase (GST)-Pak1 fusion protein, overproduced in bacteria, can be purified in an active form with glutathione affinity beads or by immunoprecipitation. Thus, other protein subunits of Pak1 are not required for its activity. In vitro protein kinase assays show that GST-Pak1 can autophosphorylate, and can phosphorylate casein as an exogenous substrate. The phenotype of the suppressed cdc17-1 cells indicates that Pak1 suppression is inefficient and does not restore the wild-type phenotype. Pak1 suppression requires Rad9 function, but Pak1 does not affect Rad9 function. Overexpression of PAK1 does not enhance the expression of the POL1 gene. Pak1 may function by modifying and partially stabilizing thermolabile DNA polymerases, perhaps during DNA repair, because pak1 mutant cells are caffeine sensitive. PMID- 9341679 TI - The Listeria monocytogenes iap gene as an indicator gene for the study of PrfA dependent regulation. AB - The iap gene of Listeria monocytogenes encodes the extracellular protein p60, which possesses a murein hydrolase activity necessary for septum separation. We constructed L. monocytogenes EGD strains harbouring plasmids that carry the iap gene under the control of the PrfA-regulated promoters of the L. monocytogenes genes hly, mpl, and actA. After insertional inactivation of the chromosomal iap gene in L. monocytogenes EGD, p60 synthesis was strictly dependent on PrfA. Elevated temperature (40 degrees C) enhanced synthesis of p60 in L. monocytogenes when the iap gene was under the control of the hly promoter; this appeared to be associated with increased synthesis of PrfA at this temperature. Synthesis of p60 in L. monocytogenes was significantly lower when the iap gene was placed under the control of the actA or the mpl promoter. Transcription of the iap gene was repressed in L. monocytogenes in the presence of PrfA when iap expression was under the control of the prfA promoter P2. Under the control of the hly promoter the gene produced low levels of secreted p60 in the presence of low amounts of PrfA, and this in turn led to the generation of long listerial cell filaments consisting of bacteria that had failed to separate. Overexpression of p60 in the presence of high levels of PrfA caused formation of single cells, which showed reduced viability depending on the level of secreted p60. These data suggest that the iap gene may be a valuable tool for monitoring virulence gene regulation by PrfA under in vivo conditions, without disturbing the integrity of the infected host cells. PMID- 9341680 TI - Characterization of an lrp-like (lrpC) gene from Bacillus subtilis. AB - In the course of the Bacillus subtilis genome sequencing project, we identified an open reading frame encoding a putative 16.4 kDa protein. This protein shows, respectively, 34% and 25% identity with the Escherichia coli regulatory proteins Lrp and AsnC. Phylogenetic analysis suggests that it represents a new group in the AsnC-Lrp family. Sequence comparisons, as well as immunodetection experiments, lead to the conclusion that the product of this B. subtilis lrp-like gene is a bona fide Lrp protein-the first one to be detected in gram-positive bacteria. When expressed in E. coli, the B. subtilis Lrp-like protein is able to repress, by about two-fold, the expression of the ilvIH operon which is normally regulated by E. coli Lrp, indicating functional similarity in their regulatory targets. Vegetative growth of a B. subtilis lrp-like mutant is not affected in rich medium. However, the lrp-like mutation causes a transitory inhibition of growth in minimal medium in the presence of valine and isoleucine, which is relieved by leucine. This points to a possible role in regulation of amino acid metabolism. In addition, sporogenesis occurs earlier in the lrp-like mutant than in the reference strain, implying that the B subtilis Lrp-like protein plays a role in the growth phase transition. PMID- 9341681 TI - Interaction of the regulator proteins RcsA and RcsB with the promoter of the operon for amylovoran biosynthesis in Erwinia amylovora. AB - The RcsA and RcsB proteins of Erwinia amylovora and Escherichia coli were expressed in E. coli and purified. Their DNA-binding activity was examined using a 1-kb DNA region containing the putative promoter of the ams operon of Ew. amylovora, which is responsible for the biosynthesis of the exopolysaccharide amylovoran. Mobility shift assays indicated specific binding of RcsA and RcsB to a region of 78 bp spanning nucleotide positions -578 to -501 relative to the translational start of the first open reading frame of the operon. This region includes stretches of homology to E. coli sigma 70 promoter consensus sequences and to the E. coli cps promoter region. Binding of the Rcs proteins was not found at a JUMPstart consensus, typical for various promoters of polysaccharide gene clusters. DNA-binding activity was not detected for RcsA alone and only high concentrations of RcsB were able to interact with the ams promoter in our assay. The two proteins bind cooperatively at the indicated region of the ams promoter and further evidence is provided showing that the DNA-protein complex formed involves a heterodimer of RcsA and RcsB. The specific activity of RcsA, but not of RcsB, was enhanced when the protein was expressed in E. coli at 28 degrees C, relative to expression at 37 degrees C. In addition, DNA-protein complex formation is affected by temperature. The E. coli RcsA/RcsB proteins bind to the same region of the ams promoter and are able to interact with the Rcs proteins from Ew. amylovora. PMID- 9341682 TI - Single-step conjugative cloning of bacterial gene fusions involved in microbe host interactions. AB - In vivo expression technology (IVET) is a genetic strategy for isolating genes expressed in vivo. In order to full exploit this technology, it is necessary to analyse large numbers of IVET-generated gene fusions, which must be recovered from the chromosome of host bacteria. In bacteria for which transductional methods are not available, the recovery of integrated fusion plasmids is problematic and currently limits broad application of IVET. We describe a rapid, single-step, triparental conjugative approach for recovering chromosomally integrated fusion plasmids from both Pseudomonas fluorescens and Salmonella typhimurium. This simple and broadly applicable conjugative cloning system extends the utility of the IVET approach to clinically and agronomically relevant microbes and may be employed to recover non-replicating and integrated plasmids in other systems. PMID- 9341683 TI - Mitochondrial protein synthesis is not required for efficient excision of intron aI5 beta from COX1 pre-mRNA in Saccharomyces cerevisiae. AB - Splicing of the group I intron aI5 beta from the yeast mitochondrial COX1 transcript requires at least four proteins, encoded by the nuclear genes PET54, MRS1/PET157, SUV3 and MSS18. These proteins either act directly to facilitate intron aI5 beta excision, or indirectly in some manner. One possible indirect mode of action of these nuclear gene products is in stimulation of expression of a mitochondrial protein, such as a maturase, that is necessary for intron aI5 beta excision. To test this possibility, splicing of intron aI5 beta was examined in a rho-strain, which is incapable of mitochondrial protein synthesis. A quantitative RT-PCR assay was set up to compare levels of spliced COX1 mRNA present in three strains: a wild-type rho + strain; the rho-strain 7-49b-11, which retains the entire COX1 transcription unit; and a strain bearing a null mutation in the nuclear PET54 gene. The results showed that excision of aI5 beta occurs relatively efficiently in the rho-strain, and therefore does not require any mitochondrial-encoded proteins. PMID- 9341684 TI - Hemodynamic cerebral ischemia. An appeal for systematic data gathering prior to a new EC/IC trial. PMID- 9341685 TI - Stroke unit treatment. Long-term effects. AB - BACKGROUND AND PURPOSE: We have previously shown that treatment in our combined acute and rehabilitation Stroke Unit improves outcome during the first year after onset of stroke compared with stroke patients treated in general wards. The aim of the present trial was to examine the long-term effects of the stroke unit care. METHODS: In a randomized controlled trial, 110 patients with symptoms and signs of an acute stroke were allocated to the Stroke Unit and 110 to general wards. No significant differences existed in baseline characteristics between the two groups. The outcome after 5 years was measured by the proportion of patients at home, the proportion of patients in an institution, the mortality, and the functional state assessed by Barthel Index. RESULTS: After 5 years, 38 (34.5%) of the patients randomized to the Stroke Unit and 20 (18.2%) of the patients randomized to the general wards were at home (P = .006). Sixty-five (59.1%) of the patients from the Stroke Unit and 78 (70.9%) of the patients from the general wards were dead (P = .041), while 7 (6.4%) and 12 (10.9%), respectively, were in an institution (e.g., nursing home) (P = NS). Functional state was significantly better for patients treated in the Stroke Unit. CONCLUSIONS: For the first time it is shown that stroke unit care improves long-term survival and functional state and increases the proportion of patients able to live at home 5 years after the stroke. Combined acute and rehabilitation stroke units appear to be an effective way of organizing treatment for acute stroke patients. PMID- 9341686 TI - Differences in pain medication use in stroke patients with aphasia and without aphasia. AB - BACKGROUND AND PURPOSE: While individuals with stroke are known to experience pain for a variety of reasons including premorbid conditions and stroke-specific sequelae, there are some groups of individuals with stroke, who because of aphasia, are unable to express their pain. This study investigated whether there exists an association between severity of aphasia and overall pain medication use as indicated (1) by the proportion of individuals medicated according to aphasia severity and (2) by the dosage of pain medication used according to aphasia severity. METHODS: The study involved a retrospective chart review of 207 charts of patients with stroke admitted to the Jewish Rehabilitation Hospital (JRH), Laval, Canada. Patients were classified into three groups according to level of expressive aphasia: those without aphasia, those with mild-to-moderate aphasia, and those severe aphasia. Information on medications used primarily for pain management was elicited for the first 21 days and the last 5 days of hospitalization. Any substitution, increase, elimination, or addition of pain medication during hospitalization was also monitored. RESULTS AND CONCLUSIONS: While the findings indicate that pain medication prescriptions were similar for all patients, a significantly smaller number of individuals with aphasia received pro re nata (prn) "as required" pain medication when compared with those without aphasia, for the first 21 days and for the last 5 days of hospitalization at the JRH. Similarly, when daily dose was monitored for the same time periods, individuals with aphasia were found to have received less medication for pain than those without aphasia. PMID- 9341687 TI - Patients' awareness of stroke signs, symptoms, and risk factors. AB - BACKGROUND AND PURPOSE: We sought to determine knowledge at the time of symptom onset regarding the signs, symptoms, and risk factors of stroke in patients presenting to the emergency department with potential stroke. METHODS: Patients admitted from the emergency department with possible stroke were identified prospectively. A standardized, structured interview with open-ended questions was performed within 48 hours of symptom onset to assess patients' knowledge base concerning stroke signs, symptoms, and risk factors. RESULTS: Of the 174 eligible patients, 163 patients were able to respond to the interview questions. Of these 163 patients, 39% (63) did not know a single sign or symptom of stroke. Unilateral weakness (26%) and numbness (22%) were the most frequently noted symptoms. Patients aged > or = 65 years were less likely to know a sign or symptom of stroke than those aged < 65 years (percentage not knowing a single sign or symptom, 47% versus 28%, P = .016). Similarly, 43% of patients did not know a single risk factor for stroke. The elderly were less likely to know a risk factor than their younger counterparts. CONCLUSIONS: Almost 40% of patients admitted with a possible stroke did not know the signs, symptoms, or risk factor of a stroke. Further public education is needed to increase awareness of the warning signs and risk factors of stroke. PMID- 9341688 TI - Is the EuroQol a valid measure of health-related quality of life after stroke? AB - BACKGROUND AND PURPOSE: The EuroQol measures aspects of quality of life that are highly relevant to stroke patients. It is short and simple and many stroke patients can complete the form without help. However, its validity has not been adequately assessed after stroke. We therefore assessed its concurrent and discriminant validity in a group of prospectively studied stroke survivors. METHODS: We assessed the validity of the EuroQol in a series of 152 patients with stroke who were all visited by a study nurse. The nurse gave the patients the EuroQol, the Frenchay Activities Index, a visual analogue pain scale, and the Hospital Anxiety and Depression Scale in the form of questionnaires to be self completed where possible. The nurse interviewed the patient directly to assess disability using the Office of Population Censuses and Surveys Disability scale and Barthel Index. RESULTS: The nurse assessed 152 patients; of these 92 were able to complete the EuroQol without help, the remaining 60 could only be assessed by interview. The EuroQol had reasonable concurrent validity; median scores on the relevant standard instruments varied significantly (and in the appropriate direction) for groups defined by their response to the relevant EuroQol domain. The EuroQol had reasonable discriminant validity since the responses enabled separation between patients with differing stroke syndromes and stroke severities. Accuracy for predicting outcome after stroke was good for both self-completed and interview-completed questionnaires. CONCLUSIONS: The EuroQol appears to have acceptable concurrent and discriminant validity for the measurement of health-related quality of life after stroke. It may be administered by either a questionnaire for self-completion in patients with mild to moderate stroke or by interview in patients with significant motor deficits. PMID- 9341689 TI - Are proxy assessments of health status after stroke with the EuroQol questionnaire feasible, accurate, and unbiased? AB - BACKGROUND AND PURPOSE: It is often difficult to determine the health-related quality of life (HRQoL) of stroke patients because physical and cognitive problems limit their ability to complete complex questionnaires. A proxy, such as a family member or caregiver, may be able to give an estimate of the patients' health status. We therefore examined the agreement between the HRQoL as assessed by a series of patients and that assessed by their proxies. METHODS: We studied the validity of the EuroQol in a series of 152 patients from our prospective registry of patients with first (or recurrent) stroke. We asked patients to ensure that a friend or relative (a proxy) who knew them well was available at the time of the interview. We asked each proxy to complete a EuroQol questionnaire independently on behalf of the patient. RESULTS: Proxies completed forms for 130 patients (86%). Agreement between responses from the patients and those from their proxies was better for patients who were able to self-complete the EuroQol than for patients who required the EuroQol to be administered by interview. For both groups, agreement was best for the self-care domain and worst for the domain that assessed psychological outcome. For the more severely affected patients, agreement was only fair for the pain and social functioning domains and no better than chance alone for the psychological functioning domain (kappa = 0.05, 95% confidence interval, 0 to 0.43). Patients tended to rate their own health status as better than their proxies did (P < .05). CONCLUSIONS: We found moderate agreement between responses from patients and those from their proxies for the more directly observable domains of the EuroQol. Proxy agreement was less good for the more subjective domains. In health surveys, allowing responses by a proxy increases response rate. However, the disadvantages inherent in the use of proxy responses must be considered carefully. In general, some domains of HRQoL information obtained from a proxy may be sufficiently valid and unbiased to be useable in most types of trials and surveys. PMID- 9341690 TI - Use of the Health Utilities Index with stroke patients and their caregivers. AB - BACKGROUND AND PURPOSE: Few studies currently assess the health-related quality of life of individuals following a stroke. One of the major challenges of assessing quality of life is the high likelihood that after a stroke a patient will not be able to complete such an assessment. One practical solution is to have a family caregiver complete the assessment on behalf of these individuals. This current pilot study examined the interrater reliability of having family caregivers complete the Health Utilities Index (HUI) on behalf of stroke patients. METHODS: A total of 74 patients who experienced an ischemic stroke and 37 family caregivers completed the interviewer-administered HUI (data were available for 33 pairs). The HUI is designed to produce a single summary measure of health-related quality of life, the global multiattribute utility score, as well as descriptive information on each of its attributes. Interrater reliability was measured by evaluating the percent agreement, Cohen's kappa statistics, intraclass correlation coefficients (ICCs), Pearson's R correlations, and paired t tests between the patient and caregiver responses. RESULTS: In most instances interrater reliability was acceptable, with values suggesting moderate to high agreement. The mean global multiattribute utility scores for the HUI 2 were identical for patients and caregivers (0.64 +/- 0.29), with an ICC of .72. A preponderance of patients reported decrements in several attributes of the HUI. CONCLUSIONS: These data indicate a substantial decrement in functioning in stroke patients and suggest that family caregivers can complete the HUI reliably when patients are unable to do so. PMID- 9341691 TI - Cerebral angiography practices at US teaching hospitals. Implications for carotid endarterectomy. AB - BACKGROUND AND PURPOSE: Although several clinical trials of carotid endarterectomy (CE) have been carried out in the last decade, the methods for angiographic measurement of carotid stenosis have not been standardized. How one measures carotid stenosis may affect the applicability of clinical trial results. We sought to obtain information on cerebral angiography practices at teaching hospitals in the United States. METHODS: We surveyed hospitals with an accredited radiology residency program. RESULTS: Of the 200 radiology program directors who were sent the survey, 97 responded. The angiographic complication rate was known in 68 of 97 medical centers and averaged 0.6%. The most common method being used for measurement of carotid stenosis is the NASCET method (70%). Forty-two of 97 program directors reported a decrease in the volume of angiography being performed. Of these 42, one third reported that CE was commonly being performed on the basis of noninvasive tests alone. CONCLUSIONS: The angiographic complication rate at American teaching hospitals is within the "acceptable" range. The NASCET method of stenosis measurement is the most popular among academic radiologists. The volume of cerebral angiography appears to be decreasing. How these data compare with community hospitals without an accredited radiology residency program warrants further study. PMID- 9341692 TI - Prevalence of stroke and stroke-related disability. Estimates from the Auckland stroke studies. AB - BACKGROUND AND PURPOSE: To provide estimates of the prevalence of stroke and stroke-related disability for international comparisons and for planning purposes. METHODS: Estimates of prevalence were derived from two population-based studies conducted 10 years apart in Auckland, New Zealand. The first, carried out in 1981, included information on survival and stroke-related disability to 14 years after stroke, and the second, undertaken in 1991 to 1992, included this information up to 3 years after stroke. An actuarial model was developed that took into account changes in incidence, long-term survival, and population structure. RESULTS: Overall, it was estimated that 7491 people (3793 men and 3698 women) living in Auckland (total population 945,000) in 1991 had experienced a stroke at some stage in the past. This represents an age-standardized rate of 833 per 100,000 (991 per 100,000 in men and 706 per 100,000 in women) in the population aged 15 years and older. When only those who have made an incomplete recovery are considered, prevalence falls to 461 per 100,000. Of this group, one third (173 per 100,000 population 15 years and older) required assistance in at least one self-care activity. CONCLUSIONS: Usual estimates of stroke prevalence, which include all people who have ever experienced a stroke, may overestimate by almost twofold the prevalence of stroke-related disability, since many have either recovered or have no continuing dependency related to stroke. Overall prevalence does not provide information with sufficient precision for planning and purchasing ongoing services for stroke patients. PMID- 9341693 TI - Trends in stroke incidence. The Copenhagen City Heart Study. AB - BACKGROUND AND PURPOSE: Stroke incidence has increased in some countries and decreased in others. After 20 years of intensive antihypertensive treatment the latter could be expected, and we have evaluated the sex-specific temporal trends in stroke incidence using 17 years of follow-up in the Copenhagen City Heart Study. SUBJECTS AND METHODS: Our cohort comprised 19,698 subjects living in Copenhagen, Denmark. They were invited for health examinations in the following time periods: 1976 through 1978, 1981 through 1983, and 1992 through 1994. Trends are presented for all persons who attended at least one of the two first examinations as well as the total cohort including nonresponders. Subjects between 45 and 84 years of age were followed from March 1, 1976 until March 1, 1993. Changes in age-specific stroke incidence were calculated by means of Poisson regression analysis. RESULTS: For subjects aged 45 to 64 years, no significant trends were observed, with an annual incidence rate ratio of 1.00 (95% confidence interval [CI], 0.97 to 1.03) and 1.04 (95% CI, 0.99 to 1.08) for men and women, respectively. In subjects aged 65 to 84 years a significant decrease in stroke incidence was found in men, whose annual rate ratio was 0.97 (95% CI, 0.95 to 0.99), but not in women, whose annual rate ratio was 0.98 (95% CI, 0.95 to 1.00). Throughout four observed periods the stroke incidence among men remained significantly higher than that for women. CONCLUSIONS: During the period from 1976 to 1993 there has been a decline in incidence of stroke in men and women aged 65 to 84 years that was significant only in men, whereas no changes were found for persons aged 45 to 64 years. PMID- 9341694 TI - Familial history of stroke and stroke risk. The Family Heart Study. AB - BACKGROUND AND PURPOSE: Although familial history of stroke is generally perceived to be an important marker of stroke risk, very few epidemiological studies have been published to address this hypothesis. We sought to examine whether familial history of stroke is associated with the prevalence of stroke in the Family Heart Study, a National Heart, Lung, and Blood Institute-supported multicenter study of the familial, genetic, and nongenetic determinants of cardiovascular disease in populations. METHODS: The personal and familial histories of stroke were assessed in 3168 individuals (probands) who were at least 45 years old and 29,325 of their first-degree relatives with the use of a standardized questionnaire. RESULTS: The age-, ethnicity-, and sex-adjusted stroke prevalences were 4.8%, 4.9%, and 3.9% in probands with a positive familial, paternal, and maternal history of stroke, respectively, in comparison with 2.0% in probands without any positive familial history (P < .01). The age-, ethnicity-, and sex-adjusted odds ratios (95% confidence interval) of stroke were 2.00 (1.13, 3.54) for a positive paternal and 1.41 (0.80, 2.50) for a positive maternal history of stroke. Additional statistical adjustment for the proband's history of elevated cholesterol level, cigarette smoking status, history of coronary heart disease, hypertension, and diabetes did not alter the associations. A similar pattern was seen for African Americans and European Americans. CONCLUSIONS: The increased risk of stroke among persons with a positive familial history of stroke compared with those without a familial history of stroke is consistent with the expression of genetic susceptibility, a shared environment, or both in the etiology of stroke. PMID- 9341695 TI - Risk factors for stroke due to cerebral infarction in young adults. AB - BACKGROUND AND PURPOSE: Stroke in the young is particularly tragic because of the potential for a lifetime of disablement. More than 10% of patients with stroke due to cerebral infarction are aged 55 years or younger. While a number of studies have addressed the issue of stroke mechanism in the young, quantitation of risk factors has rarely been undertaken. Given the importance of risk factor assessment in primary prevention, we aimed to assess this using case-control methodology in a hospital-based series and community-based control subjects. METHODS: A total of 201 consecutive patients with first-onset stroke due to cerebral infarction aged 15 to 55 years (mean, 45.5 years) were accrued from four teaching hospitals during 1985 to 1992 and compared with their age- and sex matched neighborhood controls. Information concerning potential risk factor exposure status was collected by structured questionnaire at interview. Stroke risks were estimated by calculating the odds ratios with multivariate logistic regression. RESULTS: Significantly increased risk of stroke was found among those with diabetes (odds ratio, 11.6 [95% confidence intervals, 1.2 to 115.2]), hypertension (6.8 [3.3 to 13.9]), heart disease (2.7 [1.1 to 6.4]), current cigarette smoking (2.5 [1.3, 5.0]), and long-term heavy alcohol consumption (> or = 60 g/d) (15.3 [1.0 to 232.0]). However, heavy alcohol ingestion (> or = 60 g) within 24 hours preceding stroke onset was not a risk factor (0.9 [0.3 to 3.4]). CONCLUSIONS: Diabetes, hypertension, heart disease, current smoking, and long term heavy alcohol consumption are major risk factors for stroke in young adults. Given that the majority of these factors are either correctable or modifiable, prevention strategies may have the potential to reduce the impact of stroke in this age group. PMID- 9341696 TI - Carotid atherosclerosis in men with low levels of HDL cholesterol. AB - BACKGROUND AND PURPOSE: A low HDL cholesterol (HDL-C) frequently occurs in conjunction with a desirable LDL cholesterol (LDL-C) and is a risk factor for coronary heart disease (CHD). Additionally, the presence of carotid atherosclerosis is a strong and independent predictor of morbidity and mortality in patients with CHD. This article describes the prevalence and correlates of sonographically detected carotid atherosclerosis in men with low levels of HDL-C and CHD but without elevated levels of LDL-C or total cholesterol. METHODS: High resolution B-mode ultrasonography was used to quantify intima-media wall thickness (IMT) in the common and internal carotid arteries and at the carotid artery bifurcation in 202 randomly selected male veterans with CHD and low levels of HDL-C who are participating in the VA HDL Intervention Trial. Ultrasonographic measurement of carotid artery wall stiffness was determined in a subset of 94 of these individuals. RESULTS: The mean maximum and single greatest carotid artery IMT measurements were 1.41 and 2.58 mm, respectively. The prevalence of ultrasound-detected carotid atherosclerosis as defined by a mean maximum IMT > or = 1.3 mm was 58.9% and by single maximum IMT > or = 1.5 mm was 87.1%. IMT was associated with increased age, lower extremity arterial disease, systolic blood pressure, and ultrasonographically measured carotid artery stiffness. CONCLUSIONS: Men with low levels of HDL-C and CHD but without elevated LDL-C or total cholesterol have a very high prevalence of ultrasound-detected carotid artery atherosclerosis. PMID- 9341697 TI - Cerebral aneurysms in the elderly in Yamaguchi, Japan. Analysis of the Yamaguchi Data Bank of Cerebral Aneurysm from 1985 to 1995. AB - BACKGROUND AND PURPOSE: The number of elderly people is markedly increasing in Japan. We have investigated the epidemiology and management outcome of cerebral aneurysms in elderly patients aged > or = 70 years. METHODS: A total of 3100 patients were enrolled in the Yamaguchi Data Bank of Cerebral Aneurysm between 1985 and 1995. Of these, 598 with ruptured cerebral aneurysms and 120 with unruptured cerebral aneurysms were elderly (ie, aged > or = 70 years). RESULTS: The number of elderly patients with cerebral aneurysms has markedly increased since 1991, and in 1995 approximately 30% of all patients with cerebral aneurysms were elderly. In cases of ruptured cerebral aneurysms, the proportion of patients with severe neurological grade did not change and that with an unfavourable outcome did not decrease throughout the 11 years. The proportion of patients with severe neurological grade in the elderly group was higher than in the younger group (< 70 years), and the management outcome of elderly patients for each neurological grade on admission was worse than that of younger patients (P < .01). However, the incidence rate of symptomatic cerebral vasospasm and rebleeding was the same for the two age groups. Eventually, 60.4% of all elderly patients with ruptured cerebral aneurysms had an unfavorable outcome. In cases of unruptured cerebral aneurysms, 63.3% of the selected elderly patients were surgically treated, and the surgical morbidity and mortality rates were 26.3% and 4.0%, respectively. These rates were nonsignificantly higher than those for younger patients. CONCLUSIONS: The number of elderly patients with cerebral aneurysms has markedly increased in Yamaguchi. Because of the unsatisfactory management outcome of ruptured cerebral aneurysms and surgical outcome of unruptured cerebral aneurysms in elderly patients during the 11-year period, we propose the treatment of unruptured cerebral aneurysms at a younger age and the use of a screening system to detect these subjects. PMID- 9341698 TI - Subcortical silent brain infarction as a risk factor for clinical stroke. AB - BACKGROUND AND PURPOSE: No prospective studies have examined the rate of symptomatic ischemic or hemorrhagic stroke in patients with subcortical silent brain infarction (SSBI) who were otherwise neurologically normal at entry into the study. This report investigates SSBI, detected by MRI, as a clinical stroke risk factor. METHODS: MRI scans were performed in 933 neurologically normal adults (30 to 81 years; mean age, 57.5 +/- 9.2 years) without history of cerebrovascular diseases who received our health screening of the brain 1 to 7 years before investigation. We obtained information of their clinical stroke onset through sending out a questionnaire for subjects. We detected SSBI (focal T2 hyperintensities larger than 3 mm with correlative T1 hypointensity), FWT2HL (focal white matter T2 hypertensity lesions similar to SSBI but without correlative T2-hypointensity), and PVH (periventricular hyperintensity) by MRI. Age, sex, family history of stroke, history of hypertension, diabetes mellitus, lipids, hematocrit, blood pressure, fasting blood sugar, smoking, alcohol habits, ischemic changes on electrocardiogram, and sclerotic changes of retinal arteries were included in the analysis. RESULTS: Incidence of SSBI was 10.6% in all subjects. No cortical infarct was detected in this series. Multiple logistic regression analysis showed that hypertension (odds ratio [OR], 4.07; 95% CI, 2.57 to 6.45), diabetes (OR, 2.41; 95% CI, 1.20 to 4.85), alcohol habits > or = 58 g/day (OR, 2.58; 95% CI, 1.50 to 4.45), retinal artery sclerosis (OR, 2.14; 95% CI, 1.32 to 2.38), and age (OR, 1.77; 95% CI, 1.32 to 2.38) were significant and independent risk factors for SSBI. For FWT2HL, hypertension (OR, 4.49; 95% CI, 2.54 to 7.96) and age (OR, 2.08; 95% CI, 1.45 to 3.00) were also independent risk factors. Risk factors for PVH were age (OR, 3.46; 95% CI, 2.23 to 5.36), hypertension (OR, 3.06; 95% CI, 1.62 to 5.78), and retinal artery sclerosis (OR, 2.25; 95% CI, 1.02 to 4.96). We found 14 brain infarctions, 4 brain hemorrhages, and 1 subarachnoid hemorrhage during observation. Annual incidence of clinical stroke was higher in the subjects with SSBI than in those without focal lesions (10.1% versus 0.77%). ORs for clinical stroke onset were 10.48 for SSBI (95% CI, 3.63 to 30.21) and 4.81 for FWT2HL (95% CI, 1.13 to 20.58). The PVH did not relate to clinical stroke onset. CONCLUSIONS: The strong association of SSBI, FWT2HL, and PVH with hypertension suggests a common underlying mechanism (presumably small-vessel vasculopathy). The SSBI showed the most significant association for clinical subcortical stroke. The FWT2HL was also a risk factor for the stroke but was less significant than SSBI. The subjects with SSBI should be considered at high risk for clinical subcortical brain infarction or brain hemorrhage. PMID- 9341699 TI - 1H-MRS differentiates white matter hyperintensities in subcortical arteriosclerotic encephalopathy from those in normal elderly. AB - BACKGROUND AND PURPOSE: White matter hyperintensities in MRI of the brain are often seen in normal elderly subjects. Radiologically, these hyperintense regions are similar to those in symptomatic patients with subcortical arteriosclerotic encephalopathy (SAE). Our aim was to discriminate white matter hyperintensities (WMH) on MRI in patients with SAE from similar appearing changes in normal elderly. METHODS: Three groups of elderly patients were studied: symptomatic patients with WMH of SAE (n = 5); asymptomatic subjects with diffuse, confluent WMH (n = 4); and elderly control subjects (n = 5). Proton density images revealed WMH in the occipital lobes. Proton magnetic resonance spectroscopy (1H-MRS) was used to acquire spectra in these hyperintensities. Metabolite concentrations were calculated from peak areas of N-acetylaspartate (NAA), creatine (Cre), and choline (Cho). RESULTS: The NAA/Cre and NAA/Cho ratios were reduced in the SAE group compared with the two asymptomatic groups (P < .05). NAA was decreased and Cho elevated in SAE compared with control subjects (P < .05). The average volumes of WMH in the SAE group (65.5 cm3) and in asymptomatic control subjects (59.4 cm3) were similar, and greater than those of the normal control group (4.0 cm3). CONCLUSIONS: Proton MRS discriminates WMH in SAE patients from those in asymptomatic elderly, suggesting differing causes of the hyperintensities. PMID- 9341700 TI - Subcortical hypoperfusion associated with asymptomatic white matter lesions on magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: We examined whether hemodynamic and metabolic abnormalities in the cerebral white matter, basal ganglia, and thalamus are associated with asymptomatic white matter lesions (WML) depicted on MR images. METHODS: A positron emission tomographic study with H2(15)O, C15O, and 15O2 was performed in eight normal control subjects without any WML (mean +/- 1 SD age, 68.5 +/- 10.2 years) and in 15 asymptomatic subjects with WML (71.3 +/- 8.5 years) to measure regional cerebral blood flow (CBF), cerebral blood volume, oxygen extraction fraction (OEF), and oxygen metabolic rate. RESULTS: In the cerebral white matter in the asymptomatic subjects with WML, significantly lower CBF (20.3 +/- 3.9 mL/100 mL per minute; P < .05) and significantly higher OEF (0.43 +/- 0.08; P < .05) were found compared with those for control subjects (23.5 +/- 2.6 mL/100 mL per minute and 0.37 +/- 0.06, respectively). The severity of WML was not related to the magnitude of hypoperfusion. In the basal ganglia, significantly lower CBF (44.9 +/- 6.9 mL/100 mL per minute; P < .01) and significantly higher OEF (0.54 +/- 0.08; P < .01) were found in the WML group than in control subjects (70.1 +/- 12.0 mL/100 mL/min and 0.39 +/- 0.03, respectively). In the thalamus, there was no significant difference in CBF and OEF between the control and WML groups. CONCLUSIONS: Hypoperfusion of the cerebral white matter and basal ganglia in asymptomatic WML subjects may be induced by the arteriosclerosis of long penetrating medullary arteries and lenticulostriate arteries but may not be directly related to the production of WML. The role of hypoperfusion in the production of WML and acceleration of its development remains to be elucidated. PMID- 9341701 TI - Activated microglial cells are colocalized with perivascular deposits of amyloid beta protein in Alzheimer's disease brain. AB - BACKGROUND AND PURPOSE: Microglial cells are present in the center of senile plaques (SPs) in Alzheimer's disease (AD) brain. Such a localization of microglial cells suggests that they are involved in the deposition or the clearance of amyloid-beta protein (A beta) in the brain. We examined their association with another type of parenchymal A beta deposit, which is termed the perivascular deposits of A beta (PA beta). METHODS: Thick sections from AD brain were stained with a three-color immunofluorescence method that labeled A beta, activated microglial cells, and vascular endothelial cells simultaneously. RESULTS: Three-dimensional observation under a laser scanning microscope confirmed that perivascular aggregates of activated microglial cells were colocalized with PA beta. CONCLUSIONS: Microglia occur in association with both SPs and PA beta, suggesting that they play important roles in the metabolism of A beta in AD brain. PMID- 9341702 TI - Endothelin in cerebrospinal fluid and plasma of patients in the early stage of ischemic stroke. AB - BACKGROUND AND PURPOSE: Endothelin 1 (ET-1), a highly potent endogenous vasoactive peptide, exerts a sustained vasoconstrictive effect on cerebral vessels. Elevation of ET-1 in plasma has been reported 1 to 3 days after ischemic stroke. Since we assumed that a much faster and more intense response may be observed in the cerebrospinal fluid (CSF) and since an increase in concentration of ET-1 in the CSF may cause constriction of cerebral vessels and eventually influence the neurological outcome, we measured ET-1 values in the CSF within 18 hours of stroke onset and compared the values with those in the plasma. METHODS: Twenty-six consecutive patients with acute stroke were clinically evaluated according to the modified Matthew Scale and underwent two repeat CT scans. Within 5 to 18 hours of stroke onset, lumbar puncture and blood samples were concomitantly obtained and tested; ET-1 levels in CSF and plasma of these patients were analyzed by radioimmunoassay and compared with the levels of a control group of patients with no neurological disease. RESULTS: The mean CSF concentration of ET-1 in the CSF of stroke patients was 16.06 +/- 4.9 pg/mL, compared with 5.51 +/- 1.47 pg/mL in the control group (P < .001). It was significantly higher in cortical infarcts (mean, 17.7 +/- 4.1 pg/mL) than in subcortical lesions (mean, 10.77 +/- 4.1 pg/mL) (P < .001) and significantly correlated with the volume of the lesion (P = .003). The correlation between ET-1 levels in the CSF and the Matthew Scale score was less significant (P = .05). Plasma ET-1 level was not elevated in any group. CONCLUSIONS: ET-1 is found to be significantly elevated in the CSF of stroke patients during the 18 hours after stroke. No elevation was demonstrated in plasma at this time period. ET-1 may be used as an additional indicator of ischemic vascular events in the early diagnosis of stroke. The dissimilarity between the CSF and plasma ET-1 concentrations may lead also to an hypothesis that there is a vasoconstrictive effect on the cerebral vessels or a neuronal effect caused by ET-1 in the mechanism of the progression of brain ischemia. PMID- 9341704 TI - S-100 protein: serum marker of focal brain damage after ischemic territorial MCA infarction. AB - BACKGROUND AND PURPOSE: Elevations of protein S-100 (S-100) in cerebrospinal fluid and serum have been reported after cerebral infarctions. The aim of our study was to evaluate the time course of serum S-100 concentrations after territorial middle cerebral artery (MCA) infarctions in correlation with clinical data and prognosis. METHODS: S-100 serum levels were serially determined in 26 patients with an acute infarction in the territory of the MCA at day 0 (within 12 hours after onset of symptoms), day 1 (24 hours after stroke onset), and days 2, 3, 4, 5, 7 or 8, and 10 after stroke and in 26 age- and sex-matched control subjects. S-100 assays were performed using a two-site radioimmunoassay technique. The clinical status was documented using the Scandinavian Stroke Scale. The functional deficit 4 weeks after stroke onset was scored by use of the modified Rankin scale. A cranial computed tomography (CCT) was performed initially and at day 4 or 5. RESULTS: Elevated concentrations of S-100 (> 0.2 microgram/L) were observed in 21 of 26 patients with MCA infarction but in none of the control subjects. S-100 levels peaked at days 2 and 3 after stroke. The S 100 concentrations in serum were significantly higher in patients with severe neurological deficits at admission, with extensive infarctions and a space occupying effect of ischemic edema as compared with the rest of the population. S 100 values were not significantly correlated with the functional prognosis. CONCLUSIONS: Presence of S-100 in serum after ischemic stroke may be due to combined leakage out of necrotic glial cells and passage through an impaired brain-blood barrier, indicating severe ischemic cell injury. Therefore, S-100 in serum can be used as a peripheral marker of ischemic focal brain damage and may be helpful for therapeutic decisions in acute ischemic stroke. PMID- 9341703 TI - S-100 protein and neuron-specific enolase concentrations in blood as indicators of infarction volume and prognosis in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Better techniques are needed to monitor infarction volume and predict neurological outcome after ischemic brain infarction. We evaluated the usefulness of serial measurements of S-100 protein versus neuron-specific enolase (NSE) in blood samples from patients with acute stroke. METHODS: Using nonisotopic sandwich immunoassays, we measured plasma concentrations of S-100 protein and NSE on admission and on days 3, 4, 7, and 14 after infarction in 44 patients (age range, 22 to 86 years; mean age, 65.1 years; 12 female, 32 male). Infarct volume was measured by volumetric CT on day 4 after ictus, and clinical outcome was assessed at discharge from hospital with the Activities of Daily Living Scale and 6 months after infarction with the Glasgow Outcome Scale. RESULTS: Peak blood levels of S-100 protein were found on day 2.5 +/- 1.3, and peak levels of NSE were found on day 1.9 +/- 0.8 after infarction. Peak plasma levels of S-100 protein correlated well with infarct volume (r = .75, P < .001) and with clinical outcome assessed with the Glasgow Outcome Scale (r = .51, P < .001). Serum levels of NSE correlated with infarct volume (r = .37, P < .05) but not with clinical outcome (r = .18, P > .05). CONCLUSIONS: The results of our study indicate that measuring blood concentrations of S-100 protein periodically in the first 10 days after cerebral infarction helps to predict infarct volume and the long-term neurological outcome more accurately than periodic measurements of blood concentrations of NSE. PMID- 9341705 TI - Transcranial Doppler ultrasound criteria for hemodynamically significant internal carotid artery stenosis based on residual lumen diameter calculated from en bloc endarterectomy specimens. AB - BACKGROUND AND PURPOSE: Transcranial Doppler (TCD) is often used in conjunction with carotid duplex ultrasonography (CDUS) to evaluate the hemodynamic significance of internal carotid artery (ICA) stenosis. We examined the sensitivity and specificity of TCD criteria for detection of a hemodynamically significant stenosis (residual lumen diameter < 1.5 mm) at the origin of the ICA. METHODS: We selected patients who underwent carotid end-arterectomy (CEA) and had preoperative TCD data available. Eighty-one patients underwent transorbital evaluation, 49 of whom also had transtemporal TCD performed. The endarterectomy specimens were removed en bloc and sectioned, and the minimal residual lumen diameter calculated by computer analysis. RESULTS: For the transorbital approach, the strongest indicators of a residual lumen diameter < 1.5 mm were reversed flow in the ipsilateral ophthalmic artery and a > 50% peak systolic velocity difference between the carotid siphons (distal ICAs) in patients with unilateral ICA origin stenosis. They were 100% specific and 31% and 26% sensitive, respectively. For the transtemporal approach in patients with a unilateral stenosis, a > 35% difference in ipsilateral middle cerebral artery (MCA) peak systolic velocity relative to the contralateral MCA or a > 50% difference in contralateral anterior cerebral artery (ACA) peak systolic velocity relative to the ipsilateral ACA were 100% specific for identifying a residual lumen diameter of < 1.5 mm. Sensitivities were 32% and 43%, respectively. Irrespective of contralateral stenosis, a > 35% difference in ipsilateral MCA peak systolic velocity relative to the ipsilateral posterior cerebral artery had a 100% specificity and a 23% sensitivity for detecting a < 1.5 mm minimal residual lumen diameter. CONCLUSIONS: Although the TCD sensitivity for detecting a hemodynamically significant stenosis is relatively low, it can be highly specific (up to 100%). We conclude that TCD enhances the specificity of highly sensitive CDUS criteria for detecting a hemodynamically significant ICA stenosis. PMID- 9341706 TI - Reproducibility of ultrasonographically determined intima-media thickness is dependent on arterial wall thickness. The Tromso Study. AB - BACKGROUND AND PURPOSE: We compared the reproducibility of B-mode ultrasonographic measurements of intima-media thickness (IMT) in various segments of the right carotid artery and examined whether measurement error was associated with IMT or cardiovascular risk factor levels. METHODS: In 1994/1995 a total of 6676 participants in the Tromso Study underwent ultrasound examination of common carotid artery IMT. Reproducibility of measurements was assessed by inviting 111 participants to a second ultrasound scan within 3 weeks of the first scan. On each occasion the subjects were examined by three sonographers. RESULTS: The mean between-observer absolute differences in IMT in the far wall of the bifurcation and the near and far walls of the common carotid-artery were 0.15, 0.10, and 0.08 mm, respectively. The corresponding within-observer differences were 0.15, 0.10, and 0.06 mm, respectively. Approximately 70% to 80% of total measurement variability was due to differences among sonographers; the rest was attributable to within-reader variability. Measurement error increased significantly with increasing IMT: the increase was more than twofold over the range of measurements. Cardiovascular risk factor levels were not associated with measurement variability when we controlled for IMT. CONCLUSIONS: We conclude that B-mode ultrasound provides reproducible estimates of the IMT in both the near and far walls of the carotid artery. Although measurement error is generally small, it increases proportionally with the level of IMT. PMID- 9341708 TI - Cerebral microembolic signals during cardiopulmonary bypass surgery. Frequency, time of occurrence, and association with patient and surgical characteristics. AB - BACKGROUND AND PURPOSE: We sought to determine the number of cerebral microembolic signals (MES) and their time of occurrence during the two most frequent types of cardiopulmonary bypass (CPB) surgery: coronary artery bypass grafting (CABG) and cardiac valve replacement (VR). Furthermore, we sought to examine the association between MES, patient characteristics, and intraoperative parameters. METHODS: Forty-two patients were studied, 15 of whom had CABG and 27 VR. Cerebral MES were detected with the use of transcranial Doppler monitoring of the right middle cerebral artery. RESULTS: Cerebral MES were detected in all patients. The number was significantly higher during VR (median, 1048) than during CABG (median, 82) (P < .001). In VR patients, 85% of the MES were detected when the heart regained effective ejection. During CABG, the highest number was detected when the aorta was cross-clamped (18%) and on release of the side clamp (13%). The numbers of MES during the period when the aorta was cross-clamped and in association with surgical procedures were not significantly different in the two patient groups. The total number of MES was inversely correlated to nasopharyngeal temperature (P < .01). CONCLUSIONS: A significantly higher number of cerebral MES were detected during VR than during CABG. The highest number occurred in VR patients when effective heart ejection was regained and in CABG patients when the aorta was cross-clamped and on release of the side clamp. The total number of MES increased at lower nasopharyngeal temperatures. Transcranial Doppler monitoring may alert the surgical team when emboli enter the cerebral circulation during CPB surgery, thus allowing preventive measures to be taken. PMID- 9341707 TI - Power Doppler imaging of carotid artery stenosis. Comparison with color Doppler flow imaging and angiography. AB - BACKGROUND AND PURPOSE: Power Doppler imaging (PDI) is a new sonographic technique that has recently been introduced for vascular application. Since the technical principles of PDI may provide increased sensitivity to visualize the continuity of blood flow in arterial stenoses, we investigated the diagnostic significance of PDI and the intermethod relationship for the measurement and classification of internal carotid artery (ICA) stenosis in comparison with both color Doppler flow imaging (CDFI) and angiography. METHODS: One hundred patients with a total of 128 ICA stenoses (50% to 69%, n = 37; 70% to 79%, n = 27; 80% to 99%, n = 64) and 12 ICA occlusions were consecutively investigated by means of PDI, CDFI, and intra-arterial angiography (n = 48). Reduction of the intrastenotic lumen was measured on longitudinal and transverse views of PDI and CDFI for the calculation of the degree of diameter and area stenosis, respectively. Angiographic stenosis was determined with the use of the North American Symptomatic Carotid Endarterectomy Trial (NASCET), European Carotid Surgery Trial (ECST), and common carotid (CC) methods. RESULTS: PDI provided significantly more excellent or good (92% versus 79%; P < .01) displays of the intrastenotic lumen than CDFI, particularly in complicated high-grade stenosis. While linear regression analysis demonstrated a high overall correlation between PDI and CDFI for diameter (r = .88; P < .001) and area stenosis (r = .79; P < .001), categorization of ICA stenosis revealed best agreement for 80% to 99% area stenoses. Since angiography frequently either underclassified (NASCET method) or overclassified (ECST, CC methods) the degree of ICA stenosis in comparison to both PDI and CDFI, the sonographic-angiographic correlation was only moderate (regression coefficients ranged from .62 to .70; P < .001). CONCLUSIONS: PDI further improves the assessment of ICA stenosis by providing better visualization of the stenotic vascular lumen than CDFI. Sonographic imaging of the stenotic plaque on both PDI and CDFI provided a direct measurement of the local degree of stenosis, while the angiographic grade of stenosis essentially depended on the method used for evaluation. PMID- 9341709 TI - Magnetic resonance angiography demonstrates vascular healing of carotid and vertebral artery dissections. AB - BACKGROUND AND PURPOSE: Dissection of the carotid and vertebral arteries is most accurately diagnosed with conventional angiography. MR techniques are sensitive for detecting the abnormalities associated with dissection but may lack specificity. We hypothesized that MR may be useful for serial monitoring of dissection and may therefore guide therapy. METHODS: All patients with angiographically proven carotid and/or vertebral artery dissection from July 1994 to June 1996 were followed for a median duration of 10.5 months. Of these 29 patients (44 vessels), 18 were concurrently evaluated with MR, and a target group of 9 patients (17 vessels) was prospectively followed with MR at 3-month intervals. RESULTS: In the 18 patients with both imaging studies at baseline, angiography revealed 30 dissected vessels while MR detected 27 (90%). In the target group of 9 patients, initial MR identified 15 of the 17 dissections diagnosed with angiography. Serial MR revealed complete healing in 5 vessels, improvement in 6 vessels, no change in 4 vessels, and worsening in 2 vessels. The radiographic features most likely to resolve were stenosis and mural hematoma, while occlusion and luminal irregularity tended to persist. Late ischemic events occurred in 2 patients, both with persistent MR evidence of dissection, one while subtherapeutic on warfarin therapy and the other occurring 1 week after warfarin was discontinued. CONCLUSIONS: MR is a reliable noninvasive method for following the vascular response to treatment and may guide the course of a clinical trial comparing medical therapies for carotid and vertebral artery dissection. PMID- 9341710 TI - A new method for quantitative regional cerebral blood volume measurements using computed tomography. AB - BACKGROUND AND PURPOSE: Knowledge of cerebral blood volume (CBV) is invaluable in identifying the primary cause of brain swelling in patients with stroke or severe head injury, and it might also help in clinical decision making in patients thought to have hemodynamic transient ischemic attacks (TIAs). This investigation is concerned with the development and clinical application of a new method for quantitative regional CBV measurements. METHODS: The technique is based on consecutive measurements of cerebral blood flow (CBF) by xenon/CT and tissue mean transit time (MTT) by dynamic CT after a rapid iodinated contrast bolus injection. CBV maps are produced by multiplication of the CBF and MTT maps in accordance with the Central Volume Principle: CBV = CBF x MTT. The method is rapid and easily implemented on CT scanners with the xenon/CBF capability. It yields CBV values expressed in milliliters of blood per 100 grams of tissue. RESULTS: The method was validated under controlled physiological conditions causing changes that were determined both with our technique and from pressure volume index (PVI) measurements. The two independent estimates of CBV changes were in agreement within 15%. CBV measurements using this method were carried out in normal volunteers to establish baseline values and to compare with values using the ratio-of-areas method for calculating both CBF and CBV from the dynamic study alone. Average CBV was 5.3 mL/100 g. The method was also applied in 71 patients with severe head injuries and in 1 patient with hemodynamic TIAs. CONCLUSIONS: The primary conclusions from this study were (1) the proposed method for measuring CBV accurately determines changes in CBV; (2) the MTT x CBF determinations are in agreement with the ratio-of-areas method for CBV measurements in normal volunteers and are consistent with other methods reported in the literature; (3) MTTs are significantly prolonged early after severe head injury, which when combined with the finding of decreased CBF and increased arteriovenous difference of oxygen indicates increased cerebrovascular resistance due to narrowing of the microcirculation consistent with the presence of early ischemia; and (4) CBV in the patient with TIAs was increased in the hemisphere with the occluded internal carotid artery, indicating compensatory vasodilation and probable hemodynamic cause. PMID- 9341711 TI - Acetylsalicylic acid increases tolerance against hypoxic and chemical hypoxia. AB - BACKGROUND AND PURPOSE: Treatment with acetylsalicylic acid (ASA) is established for secondary stroke prevention. Recent studies showed neuroprotection of ASA against glutamatergic excitants. The goal of this study was to investigate the time course of neuroprotection of ASA against indirect excitotoxicity by hypoxic hypoxia and chemical hypoxia. METHODS: Population spike amplitude (PSA) and ATP content were measured in hippocampal slices from untreated control animals (c slices) and slices prepared from animals pretreated in vivo with a single intraperitoneal injection of 20 mg/kg body wt ASA 1 to 48 hours before slice preparation (p-slices). RESULTS: Posthypoxic recovery of PSA was 30% in c-slices (15 minutes of hypoxia, 45 minutes of recovery). When c-slices were treated in vitro for 15 minutes with 20 mg/L ASA 30 minutes before hypoxia, posthypoxic recovery improved to 82 +/- 4% (mean +/- SE, P < .01). In p-slices, posthypoxic recovery of PSA improved in a time-dependent manner. With a time interval of 1 hour between in vivo pretreatment with ASA and slice preparation, posthypoxic recovery of PSA was 64 +/- 16% (P < .05). With time intervals of 6 hours, 24 hours, and 48 hours, posthypoxic recovery of PSA was 87 +/- 19% (P < .01), 59 +/- 12%, and 40 +/- 9%, respectively. Pretreatment with ASA in vitro or in vivo decreased the decline of ATP content during hypoxic hypoxia and chemical hypoxia (inhibition of succinic dehydrogenase by 3-nitropropionic acid). When extracellular glucose was reduced to 4 mmol/L, no difference was observed between c-slices and p-slices. CONCLUSIONS: We conclude that ASA is neuroprotective against hypoxic hypoxia and chemical hypoxia and delays the decline of intracellular ATP content. PMID- 9341712 TI - Cell death suggestive of apoptosis after spinal cord ischemia in rabbits. AB - BACKGROUND AND PURPOSE: After spinal cord ischemia, some neurons remain viable after an ischemic insult but may be at risk of dying during reperfusion. We searched for morphological and biochemical features of apoptosis, which is a mechanism of delayed neuronal death, in a rabbit model of spinal cord ischemia. METHODS: The infrarenal aorta of White New Zealand rabbits (n = 24) was occluded for 40 minutes using a loop tourniquet. Rabbits were killed after 12, 24, or 48 hours (n = 8 per group). The loop was placed but never tightened in sham-operated rabbits (n = 6). The lumbar segment of the spinal cord (L5 to L7) was used for morphological studies, including hematoxylin and eosin staining and a modified terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling (TUNEL) staining method. Electron microscopy was used to examine ultrastructural morphology. In addition, lumbar tissue was used for biochemical investigation of DNA laddering by agarose gel electrophoresis. RESULTS: After ischemia, the affected areas contained neurons with positive TUNEL staining. Positive neurons were located in laminae III to IX, although most were concentrated in the intermediate and ventral areas. Adjacent sections stained with hematoxylin and eosin exhibited ischemic cell changes (red and ghost neurons), while apoptotic bodies were also apparent. In addition, electron microscopy of ischemic tissue samples exhibited ultrastructural characteristics of apoptosis, including nuclear condensation and relatively normal organelle morphology. Finally, isolated DNA revealed a ladder on agarose gel electrophoresis, indicating DNA fragmentation into approximately 180 multiples of base pairs. CONCLUSIONS: Spinal cord ischemia in rabbits induces morphological and biochemical changes suggestive of apoptosis. These data raise the possibility that apoptosis contributes to neuronal cell death after spinal cord ischemia. PMID- 9341714 TI - Horseradish peroxidase as a histological indicator of mechanisms of porcine retinal vascular damage and protection with perfluorocarbons after massive air embolism. AB - BACKGROUND AND PURPOSE: This laboratory has previously shown that a second generation perfluorocarbon emulsion (PFE) reduces the severity of cerebral injury induced by air embolism during cardiopulmonary bypass (CPB). Horseradish peroxidase examines vascular permeability and was used in this study of the mechanisms of cellular protection afforded by the PFE. METHODS: Twenty domestic pigs underwent CPB with a prime of standard crystalloid or PFE (5 mg/kg) and crystalloid. After 10 minutes on CPB, a 5-mL/kg bolus of room air or saline (control) was delivered via the right carotid artery. The air insult was delivered as either a single bolus or double bolus. After 1 hour of CPB and 1 hour of spontaneous reperfusion, horseradish peroxidase was injected intravenously and circulated for 15 minutes. After euthanasia, both eyes were removed, and the retinas were isolated for histological analysis. RESULTS: Total length of retinal vessels exhibiting horseradish peroxidase extravasation was significantly less in PFE pigs (P < .05). Vascular spasm and red blood cell hemorrhages were unaffected by PFE. PFE pigs exhibited mild to moderate vascular nonperfusion and red blood cell sludging; crystalloid groups demonstrated severe nonperfusion and sludging. CONCLUSIONS: Histological staining with horseradish peroxidase indicated that mechanisms of cerebral air embolism include vascular endothelial leakage, vascular nonperfusion, and red blood cell sludging and hemorrhage. Pretreatment with PFE prevented some sequelae associated with massive air embolism and CPB. PMID- 9341713 TI - Superior neuroprotective efficacy of a novel antioxidant (U-101033E) with improved blood-brain barrier permeability in focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: The vascular endothelium and parenchyma of the brain have the potential to generate free radicals under pathological conditions, but it is unclear which of these two sites prevails in the production of free radicals and should be the primary target of therapeutic intervention. To clarify this issue, we compared the neuroprotective properties of a 21-aminosteroid (U-74389G) that acts on the microvasculature and a pyrrolopyrimidine (U-101033E), a novel antioxidant compound that has significantly improved potential to enter the brain parenchyma. METHODS: In Sprague-Dawley rats the middle cerebral artery was occluded for 90 minutes by an intraluminal filament. Local cortical blood flow was recorded by bilateral laser Doppler flowmetry throughout ischemia and 1 hour of reperfusion. Three groups of rats were studied: controls that received vehicle only and animals that received either U-74389G or U-101033E. Neurological examinations were performed daily, and infarct size was assessed histologically 7 days after ischemia. RESULTS: U-101033E reduced infarct volume significantly by 51%, whereas U-74389G led to a nonsignificant decrease in infarct volume. U 101033E improved neurological function immediately after ischemia, whereas U 74389G led to improvement only at the end of the observation period. Laser Doppler measurements showed no significant difference in local cortical blood flow among the treatment groups. CONCLUSIONS: We conclude that for treatment of transient focal ischemia, an antioxidant that crosses the blood-brain barrier might be superior to agents that predominantly act on the endothelium of the cerebral microvasculature. PMID- 9341715 TI - Inhibition of experimental vasospasm with anti-intercellular adhesion molecule-1 monoclonal antibody in rats. AB - BACKGROUND AND PURPOSE: Inflammation may play a role in delayed chronic vasospasm after aneurysmal subarachnoid hemorrhage. We investigated the role of intercellular adhesion molecule-1 (ICAM-1) and macrophage/granulocyte infiltration in the rat femoral artery model of vasospasm using systemic administration of a murine anti-ICAM-1 monoclonal antibody (MAb). METHODS: The femoral arteries (n = 72) in Sprague-Dawley rats (n = 36) were enclosed in latex pouches bilaterally. Autologous blood was injected into the pouch on one side, and saline was injected on the contralateral side. Chronic vessel narrowing was evaluated with the use of 29 rats, which were randomized into one of three groups for intraperitoneal injections: (1) anti-ICAM-1 MAb (2 mg/kg per dose, n = 10), (2) isotype-matched MAb (2 mg/kg per dose, n = 9), or (3) saline (n = 10), given at 3 hours and 3, 6, and 9 days after blood exposure. These rats were killed 12 days after blood exposure, and femoral artery lumen cross-sectional areas were determined by computerized image analysis. Saturation of ICAM-1 binding sites with this dosing schedule was evaluated by fluorescence-activated cell sorter (FACS) analysis of splenocytes. Immunohistochemical studies with objective cell counts were performed to evaluate macrophage/granulocyte infiltration at 24 hours in 7 rats, comparing anti-ICAM-1 MAb treatment (n = 4) with isotype-matched control MAb (n = 3). RESULTS: Animals treated with anti-ICAM-1 MAb showed a significant inhibition of arterial narrowing at 12 days (P = .0081), with lumen patency of 96.5 +/- 5.3% (mean +/- SEM), compared with 77.3 +/- 5.6% for isotype matched MAb and 72.2 +/- 5.3% for saline-treated controls. FACS analysis of splenocytes from animals treated with anti-ICAM-1 MAb confirmed saturation of ICAM-1 binding sites. Vessels treated with anti-ICAM-1 MAb showed a significant decrease in inflammatory cell infiltrates, with objective macrophage/granulocyte counts of 31.3 +/- 26.6 (mean +/- SEM) per high-powered field, compared with 171.4 +/- 30.7 for isotype-matched control MAb (P = .0027). CONCLUSIONS: Anti ICAM-1 MAb administered systemically starting 3 hours after blood exposure results in significant inhibition of chronic vasospasm in the rat femoral artery model and is correlated with a reduction in the number of infiltrating macrophages and granulocytes in the periadventitial region of blood-exposed arteries. We conclude that inflammatory changes associated with ICAM-1-mediated macrophage and granulocyte migration play an important role in the development of posthemorrhagic chronic vasospasm in this model. PMID- 9341716 TI - Rapid induction of vascular endothelial growth factor gene expression after transient middle cerebral artery occlusion in rats. AB - BACKGROUND AND PURPOSE: Vascular endothelial growth factor (VEGF) is a mitogen for endothelial cells and also has the potential to increase vascular permeability. Therefore, it may contribute to the recovery of brain cells from ischemic insult through potentiating neovascularization or may exacerbate brain damage by forming brain edema. However, the exact role of this protein in cerebral ischemia is not fully understood. We investigated temporal, spatial, and cellular profiles of the induction of VEGF gene expression after transient focal cerebral ischemia at both mRNA and protein levels. METHODS: We used a transient middle cerebral artery (MCA) occlusion model. Northern blot analysis was performed to assess the chronological pattern of induction and the impact of length of ischemia on mRNA expression. Western blot analysis was performed to ensure the selective detection of immunoreactive VEGF with an antibody. Temporal, spatial, and cellular changes of immunohistochemical VEGF expression were compared with different periods of reperfusion from 1 hour to 7 days after transient MCA occlusion. RESULTS: (1) Northern blot analysis revealed no detectable VEGF mRNA in the control brains. The mRNA became evident at 1 hour after reperfusion, peaked at 3 hours, and then decreased. The length of ischemia from 1 to 3 hours made no differences in the degree and temporal profile of the subsequent induction of VEGF mRNA. (2) Western blot analysis showed no band in the control brain, but two bands with molecular weights of 38 and 45 kD, corresponding to VEGF121 and VEGF165, were induced at 1 hour of reperfusion, peaked at 3 hours of reperfusion, and then decayed. (3) Neurons in the cerebral cortex of the MCA territory expressed VEGF at 1 hour after reperfusion with a peak at 3 hours and then diminished by 1 day. Pial cells of the MCA territory also expressed immunoreactive VEGF from 1 hour of reperfusion that was sustained until 3 to 7 days after reperfusion. CONCLUSIONS: Rapid induction of VEGF gene expression after transient MCA occlusion was demonstrated at both mRNA and protein levels. Cortical neurons and pial cells were the source of VEGF production in this model, but the temporal profiles of the induction between these cells were different. The early but dissociative induction of VEGF between neuronal and pial cells suggests different roles of the protein in their cells after transient MCA occlusion. PMID- 9341717 TI - Early detection of irreversibly damaged ischemic tissue by flumazenil positron emission tomography in cats. AB - BACKGROUND AND PURPOSE: Ligands for cerebral benzodiazepine receptors were used in the past to indicate the intactness of cortical neurons in subacute to chronic states after stroke and thus to differentiate among brain regions with complete or incomplete infarction and with functional deactivation. For planning acute interventional therapy, however, a marker of irreversible damage in early ischemia is needed. We studied the applicability of [11C]flumazenil (FMZ) for differentiation between tissue with and without potential of recovery in the first hours after focal experimental ischemia. METHODS: In 11 cats, cerebral blood flow, cerebral metabolic rate for oxygen, oxygen extraction fraction, and FMZ binding were studied repeatedly by positron emission tomography before, during, and up to 12 hours after transient middle cerebral artery occlusion (MCAO) (30 minutes in 2, 60 minutes in 7, and 120 minutes in 2 cats, respectively). Development of the defects in energy metabolism were compared with the defects in FMZ binding (2 to 3 hours and 8 to 9 hours after MCAO), with the pattern of disturbed glucose metabolism (determined 12 hours after MCAO), and with the size of the infarcts (determined approximately 15 hours after MCAO). RESULTS: Irrespective of the level of reperfusion, defects in FMZ binding (2 to 3 hours after MCAO) were closely related to areas with severely depressed oxygen consumption and predicted the size of the final infarcts, whereas preserved FMZ binding indicated intact cortex. Depression of glucose metabolism was in all animals larger than the defects in FMZ binding and the infarcts, indicating functional deactivation of brain areas beyond the permanent morphological damage. In addition, FMZ distribution within 2 minutes after injection was significantly correlated to flow and yielded reliable perfusion images. CONCLUSIONS: The reduction of FMZ binding early after focal ischemia reflects irreversible neuronal damage that otherwise only can be detected by multitracer studies. Our experimental data and first clinical applications suggest that FMZ has potential as an indicator of developing infarction. Since FMZ distribution additionally images perfusion, this tracer might be useful for the selection of patients who would benefit from acute therapeutic intervention. PMID- 9341719 TI - Correlation between motor impairment and infarct volume after permanent and transient middle cerebral artery occlusion in the rat. AB - BACKGROUND AND PURPOSE: There have been a number of recent reports describing the relationship between ischemic damage and various behavioral and functional measures, although there have been few studies that have demonstrated a direct correlation between functional impairment and lesion volume. The purpose of the present study was to assess functional outcome by measurement of motor impairment and to determine whether this correlated to a range of infarct volumes induced by varying the duration of focal ischemic insult in the rat. METHODS: Male Sprague Dawley rats were subjected to 0, 30, 60, of 120 minutes or permanent middle cerebral artery (MCA) occlusion by the intraluminal filament technique. Motor impairment was assessed by the accelerating rota-rod and grid-walking tests, and the brains were perfusion-fixed for histological determination of infarct volume and brain swelling 24 hours after MCA occlusion. RESULTS: Marked impairment in performance of both motor tests was recorded in the 60-minute, 120-minute, and the permanent MCA occlusion groups when compared with sham-operated rats. There were significant correlations between regional infarct volume, brain swelling, and all behavioral measurements (all r2 > .5, P < .001). CONCLUSIONS: The rota rod and grid-walking tests of motor performance provide quantitative, objective, and reproducible measures of functional impairment of rats following an ischemic insult. These impairments correlate directly with infarct volume and provide information integral to future studies evaluating the effects of potential neuroprotective agents. PMID- 9341718 TI - Extracellular nucleotide-induced [Ca2+]i elevation in rat basilar smooth muscle cells. AB - BACKGROUND AND PURPOSE: Extracellular nucleotides play an important role in the regulation of vascular tone and may be involved in cerebral vasospasm after subarachnoid hemorrhage. The objective of this study was to investigate the receptor subtypes for nucleotides and their mechanisms of [Ca2+]i mobilization in cerebral vasculature. METHODS: Rat basilar smooth muscle cells were isolated by an enzymatic method. [Ca2+]i response, a large transient peak followed by a slowly decaying plateau. The potency of nucleotides to raise [Ca2+]i was ATP gamma S > or = UDP > or = ATP approximately UDP approximately TTP, indicating that P2u receptors were expressed in the rat basilar smooth muscle cells. The effect of UTP to release Ca2+ from internal stores was reduced by pertussis toxin, by the phospholipase C inhibitor 2-nitro-4-carboxyphenyl N,N diphenylcarbamate, and by the Ca(2+)-pump inhibitor thapsigargin. The Ca2+ entry induced by UTP was partially attenuated by the receptor-operated Ca2+ channel blocker SK&F96365 and by the voltage-dependent Ca2+ channel blocker verapamil. P2 receptor antagonists suramin and, at higher concentrations, pyridoxal-phosphate-6 azophenyl-2',4'-disulphonic acid reduced the effect of UTP. CONCLUSIONS: The results are the first demonstration that nucleotides activate G protein-coupled P2u receptors to mobilize [Ca2+]i in rat basilar smooth muscle cells. PMID- 9341720 TI - Biophysical mechanisms of stroke. AB - BACKGROUND: Stroke is the third leading cause of death and the leading cause of long-term disability in the United States. Although a host of genetic, biochemical, physiological, anatomic, and histological factors have been implicated, to varying degrees, in the pathogenesis of stroke, biophysical factors are believed to play a significant role in the development, diagnosis, and therapy of stroke. The purpose of this review article is to identify, describe, and illustrate these causes and biophysical and hemodynamic mechanisms predisposing a person to stroke, which often form the basis for novel methods of diagnosis and therapy. SUMMARY OF REVIEW: This mini-review begins by describing the physical principles that govern the flow of blood through normal and stenosed carotid artery bifurcations. In addition to the tortuosity, curvature, and tensile forces of the carotid artery bifurcation, the effects of biophysical phenomena from flowing blood such as viscous forces, pressure forces, velocity, kinetic energy, momentum, impulse, shear stress, and vibrational displacements exerted by the flowing blood on the vessel wall are conducive to abnormal flow behavior and patterns, degrading the vessel wall and creating the potential for stroke. CONCLUSIONS: Recent advances in the treatment of stroke are based on increasing knowledge of its underlying biophysical mechanisms, as well as on better-publicized advances in imaging instrumentation and procedures for the management and treatment of patients. PMID- 9341721 TI - Intracranial aneurysm rupture presenting as delayed stroke secondary to cerebral vasospasm. AB - BACKGROUND: Days after aneurysmal subarachnoid hemorrhage (SAH), cerebral vasospasm can result in the delayed appearance of ischemic neurological deficit identical to that produced by other causes of stroke. Despite the well-described, "classic" presentation of SAH, up to 25% of patients are initially misdiagnosed, and the initial hemorrhage from a ruptured aneurysm will not always bring the patient to medical attention. CASE DESCRIPTIONS: We report our experience with two patients who presented with signs and symptoms of ischemic stroke resulting from cerebral vasospasm that followed unrecognized rupture of a brain aneurysm. In one case, it was the recent complaint of significant headache and a prior history of SAH that led to the correct diagnosis. In the other case, a major rebleed occurred before the accurate diagnosis was recognized. CONCLUSIONS: It is critical to make the correct diagnosis of stroke due to vasospasm so that appropriate treatment can be instituted, thrombolytic and anticoagulant therapy can be avoided, and the unsecured aneurysm can be obliterated to prevent potentially catastrophic rebleeding. PMID- 9341722 TI - Fou rire prodromique heralding a left internal carotid artery occlusion. AB - BACKGROUND: Four rire prodromique, described as pathological laughter preceding the onset of an apoplectic attack, is a rare phenomenon. CASE DESCRIPTION: A 61 year-old man manifested pathological laughter before a sudden right hemiplegia. MRI showed a left lenticular and caudate nucleus infarct with involvement of the external capsule and prerolandic area. MRA revealed a left internal carotid and middle cerebral artery occlusion. CONCLUSIONS: The clinicoanatomic correlates of this phenomenon are discussed. PMID- 9341723 TI - Symptomatic carotid artery occlusion. A reappraisal of hemodynamic factors. AB - BACKGROUND: Over the last several years evidence has accumulated that in addition to embolism, a compromised cerebral blood flow may play an important role in causing transient ischemic attacks and ischemic stroke in patients with occlusion of the internal carotid artery. This evidence is found in both clinical features and ancillary investigations, particularly measurements of cerebral blood flow. SUMMARY OF REVIEW: On the basis of 20 follow-up studies in patients with transient ischemic attacks or minor ischemic stroke associated with an occluded carotid artery, the annual risk of stroke was 5.5% (95% confidence interval [CI], 5.0% to 6.0%), and that of ipsilateral stroke (distinguished in 11 of the 20 studies) was 2.1% (95% CI, 1.6% to 2.8%). Patients with a compromised cerebral blood flow as measured by positron emission tomography, single-photon emission CT, transcranial Doppler, or stable xenon CT (six studies) have an even higher annual risk of stroke (all strokes: 12.5%; 95% CI, 8.9% to 17.6%; ipsilateral stroke: 9.5%; 95% CI, 6.4% to 14.0%). CONCLUSIONS: Because a compromised cerebral blood flow may be an important causal factor in patients with symptomatic carotid artery occlusion, medical and surgical options for treatment are reviewed in this light. PMID- 9341725 TI - Diagnosis of cerebral amyloid angiopathy: sensitivity and specificity of cortical biopsy. PMID- 9341724 TI - Transcranial Doppler ultrasound in severe carotid artery stenosis. PMID- 9341739 TI - Inhibition of IL-10 protein synthesis induces major histocompatibility complex class II gene expression in class II-deficient patients. AB - Major histocompatibility complex (MHC) class II deficiency is an inherited autosomal recessive combined immunodeficiency, characterized by a lack of constitutive expression of the human leukocyte antigen (HLA) class II genes. The patients investigated in this study are histoidentical twin brothers with a new phenotype in MHC class II deficiency. Examination of HLA-D locus genes in their fractionated peripheral mononuclear cells (MNCs) revealed an unusual and uncoordinated mRNA pattern. Here we analyzed the distribution of pro- and anti inflammatory cytokines expressed in these patients' adherent and nonadherent MNCs. We show that gene expression of IL-1 alpha, IL-1 beta, IL-6, granulocyte colony-stimulating factor, and IL-10 was induced in both cell fractions, whereas increased mRNA levels of interferon-gamma and the inducible nitric oxide synthase were exclusively detected in the patients' nonadherent MNCs. As IL-10 is known to be able to downregulate transcription of MHC class II and expression of IL-10 in the patients' MNCs was increased, we investigated the regulatory function of this cytokine. Interestingly, inhibition of IL-10 protein synthesis with IL-10 specific antisense oligonucleotide DNA (IL-10-AS-ODN) induced HLA-D locus genes in these MHC class II-deficient patients. Exposure of the nonadherent cell fraction to IL-10-AS-ODN resulted in a profound induction of a previously absent DR beta 1 and DP alpha gene expression. HLA-DQ beta mRNA levels, however, were increased in both the adherent and the nonadherent MNC population. Albeit expression of HLA-D locus genes was inducible via inhibition of IL-10 translation, surface expression of HLA class II antigens on the patients' MNCs was essentially negative. The data presented support the concept of a coordinated network of pro- and anti-inflammatory cytokine regulation and this network obviously has a significant role in the cell-type-specific regulation of MHC class II expression. PMID- 9341740 TI - Regulation of the expression of the soluble and membrane forms of the murine IL-4 receptor. AB - The actions of interleukin-4 (IL-4) in vivo are likely to be positively influenced by the expression of membrane IL-4 receptors (mIL-4R) on target cells and negatively by the concentration of soluble IL-4 receptors (sIL-4R) in the extracellular environment. Inasmuch as the two forms of the mouse IL-4R are differentially encoded by alternatively spliced mRNA transcripts, the purpose of this work was to determine how their expression is regulated by IL-4 and T cell activation and whether there is preferential expression of one type of transcript over the other. In this study, the expression of sIL-4R and mIL-4R transcripts was analyzed by a semiquantitative RT-PCR method in resting and mitogen-activated splenic cells. Irrespectively of the state of cell activation, IL-4 up-regulated the levels of both types of mRNA with similar kinetics and dose-response curves. In contrast, ConA failed to enhance the steady-state levels of sIL-4R or mIL-4R transcripts despite increased expression at the protein level, suggesting that sIL-4R expression is also regulated at levels other than transcription. Western blot analysis of supernatants of IL-4- and ConA-stimulated spleen cells substantiated the presence of sIL-4R molecules derived by translation of sIL-4R specific transcripts, thus confirming the importance of this mechanism for the generation of sIL-4R molecules in normal cells. These results indicate that the sIL-4R- and mIL-4R-specific transcripts are normally regulated in a parallel manner and further suggest that expression of both forms of the IL-4R is controlled at multiple levels (i.e., transcriptional and posttranscriptional). PMID- 9341741 TI - The p38 mitogen-activated protein kinase pathway in activated and anergic Th1 cells. AB - Stimulation of T cells through the TCR leads to activation of the mitogen activated protein kinase (MAPK) family members ERK (extracellular signal regulated kinase) and JNK (jun NH2-terminal kinase). These kinases act in synergy to increase the activity of the transcription factor AP-1 which is involved in the transcriptional upregulation of IL-2. Recently a third MAPK member, p38, has been identified. The effects of T cell activation on this pathway have not yet been elucidated. Using two murine Th1 clones, we demonstrate that the p38 pathway is induced upon anti-CD3 plus anti-CD28 crosslinking or PMA plus ionomycin stimulation. p38 activity was induced fully by anti-CD3 or PMA alone and is not enhanced by costimulation even at low levels of TCR signaling. p38 activity peaked at 20 min and was significantly decreased by 2 hr. Anergic (tolerant) Th1 cells showed decreased p38 activity as well as decreased ERK and JNK activities even though levels of these proteins remained unchanged. PMID- 9341742 TI - Costimulation with dexamethasone and prostaglandin E2: a novel paradigm for the induction of T-cell anergy. AB - In this report, data are presented which indicate that anti-CD3 mAb-stimulated human peripheral blood T-cells treated with both dexamethasone (DEX) and prostaglandin E2 (PGE2) become anergic. This anergy can be reversed by the addition of IL-2. Further, experiments were performed to investigate this T-cell anergy. The results show that addition of DEX and PGE2 to anti-CD3 mAb-stimulated T-cells inhibits the induction of p56lck but not p59fyn kinase activity nor is the tyrosine phosphorylation of PLC gamma altered appreciably. Additionally, this treatment of anti-CD3 mAb-stimulated T-cells also results in decreased tyrosine phosphorylation of ERK1, suggesting that the Ras activation pathway may be inhibited. Interestingly, the induction of T-cell anergy is reproduced when an agonist for the cAMP-independent EP3 subtype of the PGE2 receptor is substituted for PGE2. Thus, while the mechanisms responsible for the dual action of DEX and PGE2 on the induction of T-cell anergy is unknown, these data suggest that a cAMP independent mechanism may be involved. These data indicate that a state of anergy can be induced in normal human T-cells by the activation of these cells in the presence of physiologic concentrations of DEX and PGE2. PMID- 9341743 TI - Cord blood CD16+56- cells with low lytic activity are possible precursors of mature natural killer cells. AB - Human natural killer (NK) cells are defined as being membrane CD3-, CD16+, and/or CD56+ lymphocytes; however, little is known about the ontogenic development and maturational pathways of human NK cells. The functional, phenotypic, and maturational characteristics of human umbilical cord blood (CB) NK cell subsets were studied to gain insight into the ontogenic and maturational pathways of human NK cells. We have previously shown that there is a novel subset of CD16+ CD56- NK cells present in CB. Here we further demonstrate differences in the expression of the NK-associated molecules CD2, CD7, CD8, and CD25 between CB and peripheral blood (PB) NK cells and between CB NK cell subsets. Although CB NK cell subsets were deficient in or had less lytic activity against K562 cells compared to PB NK cells, CB NK cells did possess the lytic molecules perforin and granzyme B and when artificially stimulated to secrete their granules during lytic assays, were capable of lytic activity equivalent to that of PB NK cells. Regardless of differences in phenotype and function of CB NK cell subsets, short term and long-term incubation with cytokines induced functional (adult-like NK activity) and phenotypic (adult-like CD16+56+ or CD16-56+ surface antigen phenotype) maturation, respectively. Interleukin-2 (IL-2), IL-12, and IL-15, but not IL-7, interferon-gamma (IFN-gamma) nor tumor necrosis factor-alpha (TNF alpha) induced functional and phenotypic maturation of CB NK cell subsets. Interestingly, culture of CB NK cell subsets with IL-2 or IL-15 led to acquisition of predominantly a CD16+56+ phenotype, while culture with IL-12 led to acquisition of both CD16+56+ and CD16-56+ phenotypes. Both functional and phenotypic maturation were not dependent upon proliferation. Studies using neutralizing anti-IFN-gamma and anti-TNF-alpha antibodies showed that survival and phenotypic maturation upon cytokine stimulation is influenced by endogenous production of TNF-alpha but not IFN-gamma. These results demonstrate that CB NK cell subsets are functionally and phenotypically immature but are capable of maturation. Additionally, CD16+56- NK cells are implicated as possible precursors of mature CD16+56+ and CD16-56+ NK cells. PMID- 9341744 TI - Detection of CD4+CD45RO+ T lymphocytes producing IL-4 in response to antigens on Plasmodium falciparum erythrocytes: an in vitro correlate of protective immunity induced with attenuated Plasmodium falciparum sporozoites. AB - Malaria is caused by Plasmodium spp. and is one of the major infectious diseases leading to morbidity and mortality in tropical areas of the world. The model of protective immunity induced by immunization with radiation-attenuated Plasmodia sporozoites (SPZ) has become the framework for the elucidation of protective immune mechanisms and the prototype for a promising vaccine strategy. We have previously reported that although considered stage specific based on antibody and CD8+ cytolytic T lymphocyte responses directed against preerythrocytic stage antigens, in particular, the circumsporozoite protein and sporozoite surface protein 2, protective immunity induced in humans by attenuated Plasmodium falciparum SPZ may also involve CD4+ T cell responding to antigens present on parasitized red blood cells (pRBC). In this study we examined the functional role of pRBC responding CD4+ T cells by comparing in vitro pRBC-stimulated responses of CD4+ T cells from persons during preimmunity to irradiated SPZ, during induction of protection, and infection induced with SPZ. The results reported herein corroborate previously published observations that antigens associated with pRBC induce proliferative CD4+ lymphocytes responses in subjects exposed to malaria parasite-derived antigens and not malaria-naive persons; however, now we demonstrate that pRBC-proliferative CD4+ T cells did not coincide with protective immunity. Similarly, pRBC-induced IFN-gamma levels did not distinguish malaria protected from susceptible persons, although IFN-gamma was observed only in lymphocyte cultures from malaria parasite-exposed volunteers and not in lymphocyte cultures from malaria-naive persons. In contrast, we noted an increase in the IL-4-producing CD4+ T cells that also exhibited the memory phenotype, CD45RO, and an upregulated expression of CD25 in cultures from malaria protected persons as compared to malaria naive persons and subjects who became parasitemic. Hence, these observations suggest that the induction of memory CD4+ T cell subset distinguished by the expression of CD45RO and CD25 and production of IL-4 coincides with protective immune responses generated by immunization with attenuated SPZ. PMID- 9341745 TI - Spleen cell survival and proliferation are differentially altered by docosahexaenoic acid. AB - Omega-3 fatty acids have diverse health benefits that are not clearly understood. In this study we have examined the effects of the omega-3 fatty acid docosahexaenoic acid (DHA) on mitogen-activated and resting splenic lymphocytes. DHA inhibited lymphocyte proliferation, producing an apparent block or prolongation of S phase, without evidence for direct cytotoxicity. In contrast, DHA enhanced the survival of resting lymphocytes in culture without inducing cell cycling. When DHA was added at the start of culture, the survival advantage was apparent for 2 to 3 days, after which time typical lymphocyte attrition occurred. Using flow cytometry we observed that both T and B cell recoveries were increased by DHA, but there were DHA dose-dependent alterations of forward- and side scatter characteristics, with some preference for B cells, perhaps indicating altered membrane properties. Our data imply that DHA may check ongoing immune response while concurrently preserving resting lymphocytes needed for subsequent immune responses. PMID- 9341747 TI - The receptor function of CD2 in human CD2 transgenic mice is based on highly conserved associations with signal transduction molecules. AB - The activation of human T cells via CD2 in response to mitogenic monoclonal antibodies (mAbs) typically requires that one mAb is specific for an epitope within the N-terminal Ig domain of CD2 and the other for a partially hidden epitope. We have examined the proliferative response of human T cells and human CD2 (huCD2) transgenic murine T cells to two novel CD2 monoclonal antibodies, AICD2.M1 and AICD2.M2, and have partially mapped the epitopes of these and other mitogenic CD2-specific monoclonal antibodies by way of recognition of CD2:CD58 chimeric proteins possessing either the N-terminal or the membrane proximal immunoglobulin domains of CD2. To understand the molecular basis of proliferation in huCD2 transgenic murine T cells, the interactions of huCD2 with signaling proteins in murine T cells were analyzed. The transgenic huCD2 molecule was found to interact with the murine tyrosine kinases p56lck and p59fyn and the CD3 epsilon and zeta chains of the TCR/CD3 signaling complex and to coimmunoprecipitate tyrosine phosphatase activity. These molecular associations resemble the situation in human T cells and suggest that human CD2 couples to the same signal transduction pathways in humans and transgenic mice. PMID- 9341746 TI - Unmethylated CpG motifs protect murine B lymphocytes against Fas-mediated apoptosis. AB - CD40 ligand (CD40L) has been shown to increase surface Fas expression and induce B cell sensitivity to Fas-dependent CD4+ Th1 cell-mediated cytotoxicity (Th1 CMC). We investigated the role of unmethylated mitogenic CpG motifs in regulating Fas-mediated apoptosis in primary murine B cells. Unmethylated CpG motifs protected CD40L-stimulated B cells from Th1-CMC and apoptosis mediated by Fas specific antibody. Mitogenic CpG motifs downregulated Fas expression on CD40L stimulated B cells in a time-dependent fashion. These observations suggest that Fas-mediated apoptosis requires minimum upregulation of surface Fas expression and that CpG motifs protect B cells from Fas-mediated apoptosis by decreasing surface Fas expression. Thus, these results suggest a novel mechanism for induction of Fas resistance in B cells. PMID- 9341748 TI - Cholera toxin stimulates human B-cell DR antigen biosynthesis at the level of translation. AB - Cholera toxin (CT) exerts many diverse regulatory effects on cells of the immune system and is considered a potent adjuvant on gut mucosal immune responses to orally presented antigens. It has been previously described that CT induces surface DR expression in human resting B-cells. As a further step toward understanding this phenomenon, the molecular mechanisms underlying the regulation of DR expression were investigated. By the use of Western analysis, it is shown that CT increases the total levels of DR protein in highly purified human tonsillar cells. [35S]Methionine incorporation studies show that the aforementioned increase is due to de novo biosynthesis of DR protein at as early as 6 hr after CT stimulation and is completed by 24 hr. [3H]Uridine uptake experiments, nuclear transcription runoff assays, and Northern analysis show that CT does not exert its effect at a transcriptional level, indicating translational regulation. Anti-IgM, which mimics B-cell antigen binding, behaves in a manner similar to CT. The B subunit of CT (B-CT) and prostaglandin E2, either alone or in combination, do not induce DR protein biosynthesis nor do they exert any effect on the transcription of DR beta 1 gene. These results show that cAMP elevation as well as binding of B-CT to GM-1 ganglioside, by themselves, do not lead to DR protein biosynthesis suggesting that other activation pathways may be involved. PMID- 9341749 TI - Qa-1 interaction and T cell recognition of the Qa-1 determinant modifier peptide. AB - The peptide-binding properties of the nonclassical major histocompatibility complex (MHC) class 1b molecule Qa-1 were investigated using a transfected hybrid molecule composed of the alpha 1 and alpha 2 domains of Qa-1b and the alpha 3 domain of H-2Db. This allowed the use of a monoclonal antibody directed against H 2Db whilst retaining the peptide-binding groove of Qa-1b. By comparison with classical MHC class I molecules, intracellular maturation of the chimeric molecule was inefficient with weak intracellular association with beta 2 microglobulin. However, at the cell surface the hybrid molecules were stably associated with beta 2-microglobulin and were recognized by cytotoxic T lymphocyte (CTL) clones specific for the Qa-1b-presented peptide Qdm (AMAPRTLLL). A whole-cell binding assay was used to determine which residues of Qdm were important for binding to Qa-1b and CTL clones served to identify residues important for T cell recognition. Substitutions at position 1 and 5 did not reduce the efficiency of binding and had little effect on CTL recognition. In contrast, substitutions at position 9 resulted in loss of MHC class I binding. Mass spectrometric analysis of peptides eluted from immunopurified Qa-1b/Db molecules indicated that Qdm was the dominant peptide. The closely related peptide, AMVPRTLLL, which is derived from the signal sequence of H-2Dk, was also present, although it was considerably less abundant. The mass profile suggested the presence of additional peptides the majority of which consisted of eight to ten amino acid residues. Finally, the finding that a peptide derived from Klebsiella pneumoniae can bind raises the possibility that this non-classical MHC class I molecule may play a role in the presentation of peptides of microorganisms. PMID- 9341750 TI - An interleukin-7 internet for intestinal intraepithelial T cell development: knockout of ligand or receptor reveal differences in the immunodeficient state. AB - Both interleukin-7 (IL-7) and IL-7 receptor (R) gene knockout (IL-7-/- and IL-7R /-) mice were employed in order to directly investigate the importance of the IL 7 and IL-7R signaling pathway for the development of intestinal intraepithelial lymphocytes (IEL). Loss of the IL-7R-specific gene resulted in complete deficiency of the gamma delta T cell lineage with lack of V gamma 4- and V gamma 7-specific messages in the epithelium of the gastrointestinal (GI) tract in comparison to control mice of the same genetic background (approximately 40%). Disruption of the IL-7-specific gene resulted in marked, but not complete depletion of gamma delta T cells (2-3%) in IEL. Furthermore, mRNA for both V gamma 4 and V gamma 7 genes were detected in the gamma delta IEL subset of IL-7-/ mice. The subtle differences between IL-7-/- and IL-7R-/- mice suggest that although IL-7 controls most of the expansion and/or development of gamma delta IEL, another ligand binding to the IL-7R also plays a discernable role. Furthermore, alpha beta IEL developed more slowly in IL-7R-/- mice when compared with ligand knockouts; however, the frequency of IEL T cells subsequently increased with age and normal levels of CD3+ T cells expressing the alpha beta TCR were detected by 2 and 3 months of age in IL-7-/- and IL-7R-/- mice, respectively. The direct comparison of IL-7-/- and IL-7R-/- mice clearly supports the hypothesis that both IL-7 and another IL-7R binding molecule can influence the development of gamma delta T cells in the intestinal epithelium. PMID- 9341751 TI - Two separable T cell receptor signals reconstitute positive selection of CD4 lineage T cells in vivo. AB - Positive selection is an obligatory step during intrathymic T cell differentiation. It is associated with rescue of short-lived, self major histocompatibility complex (MHC)-restricted thymocytes from programmed cell death, CD4/CD8 T cell lineage commitment, and induction of lineage-specific differentiation programs. T cell receptor (TCR) signaling during positive selection can be closely mimicked by targeting TCR on immature thymocytes to cortical epithelial cells in situ via hybrid antibodies. We show that selection of CD4 T cell lineage cells in mice deficient for MHC class I and MHC class II expression can be reconstituted in vivo by two separable T cell receptor signaling steps, whereas a single TCR signal leads only to induction of short lived CD4+CD8lo intermediates. These intermediates remain susceptible to a second TCR signal for 12-48 h providing an estimate for the duration of positive selection in situ. While both TCR signals induce differentiation steps, only the second one confers long-term survival on immature thymocytes. In further support of the two-step model of positive selection we provide evidence that CD4 T cell lineage cells rescued by a single hybrid antibody pulse in MHC class II-deficient mice are pre-selected by MHC class I. PMID- 9341752 TI - Massive mammary gland infection and pregnancy-dependent mammary tumor development in mice infected neonatally with mouse mammary tumor virus (SW) but not in mice infected as adults, despite a dramatic local response. AB - Mouse mammary tumor virus (MMTV) (SW) caused a high incidence (65%) of pregnancy dependent adenocarcinomas in BALB/c(SW) mice infected as newborns by suckling their mothers' milk. These tumors were type B adenocarcinomas which developed early, at about 1 year of age. Uninfected breeding females and those infected at an age of 8 weeks by injection of virus had the same low incidence of malignant tumors (13%), and the tumors developed later (at approx. 23-24 months). The low incidence of tumors in adult-infected mice was correlated with partial infection of the mammary glands, and delayed transmission of MMTV(SW) to the offspring. Although the virus was rapidly disseminated in both types of infection, the responses of neonatally infected and adult-infected mice to MMTV(SW) infection and viral superantigen (vSAG) presentation were different. Activation by and presentation of the vSAG was impaired in mice infected neonatally, and tolerance induction by clonal deletion was delayed. Local activation was dramatic in mice infected as adults and clonal deletion followed rapidly. Although interaction between B and T cells is needed for completion of the virus life cycle and viral amplification, the strong local immune response to MMTV(SW) in adult-infected mice limits mammary gland infection, and protects them against mammary tumor development. PMID- 9341753 TI - Expression of mouse CD43 in the B cell lineage of transgenic mice causes impaired immune responses to T-independent antigens. AB - CD43 (leukosialin), a sialylated glycoprotein expressed on the surface of most hematopoietic cells, has been implicated in cell adhesion and signaling. However, its precise physiological function remains unclear. We used mouse CD43 (mCD43) immunoglobulin enhancer-transgenic (TG) mice to study the role of mCD43 in vivo. Previous work revealed that mCD43 expression on mature B cells in these mice resulted in immunodeficiency to T-dependent (TD) antigens (Ag), possibly by impairing B-T cell interactions. In the present study we have immunized the TG mice with the T-independent (TI) Ag fluorescein-(Fl) lipopolysaccharide (LPS) (TI type 1 Ag) and Fl-Ficoll (TI type 2 Ag). Surprisingly, the mCD43-Ig enhancer expressing mice were impaired in their ability to mount humoral responses to both Fl-LPS and Fl-Ficoll, and had decreased numbers of cells responding to Ag in vivo. Flow cytometric analysis was performed on peritoneal B-1 cells, a population which often plays a major role in humoral responses to TI Ag such as bacterial Ag. This analysis revealed similar B220, IgM and CD5 expression patterns for the TG and nontransgenic (NTG) B-1 cells. In addition, purified peritoneal B-1 cells from TG and NTG mice were able to respond to LPS. Stimulation of splenic B cells in vitro with Fl-LPS and Fl-Ficoll revealed that, in contrast to NTG B cell responses, TG B cell responses could not be enhanced by co-culture with T cells. However, soluble T cell factor enhancement of the TG B cell responses was normal. These data suggest that the mCD43 expression on B cells may inhibit cell interactions that are important for enhanced TI Ag responses. The anti-adhesive forces of mucins in general may thus be critical in regulating both TD and TI humoral responses. PMID- 9341754 TI - Elevated mutant frequencies in gene lacI in splenic lipopolysaccharide blasts after exposure to activated phagocytes in vitro. AB - The interaction of B lymphocytes with phagocytes is critical for shaping the humoral immune response, as well as various aspects of normal and malignant B cell development, and has therefore been studied by immunologists in great detail. However, one potential outcome of this confrontation is often neglected, namely the mutagenicity of phagocytes to B lymphocytes. We are interested in phagocyte-induced B cell mutagenesis and have conducted a feasibility study on the utility of a transgenic reporter assay to evaluate mutant frequencies in B cells that have encountered phagocytes. An in vitro co-incubation system was designed in which splenic lipopolysaccharide (LPS) blasts carrying a phage lambda derived lacI transgene were exposed to pristane-elicited peritoneal exudate cells (PEC). Mutant frequencies in LPS blasts were significantly increased (up to 6 fold) when the cells were co-incubated with PEC that had been stimulated by phorbol myristate acetate to undergo an oxidative burst. The lacI-based transgenic mutation assay proved also useful for assessing mutagenicity in vivo, as demonstrated by the detection of elevated mutant frequencies in the spleen (3 fold) and the inflammatory granuloma (4.7-fold) obtained from pristane-treated mice. We propose to utilize the lacI-based transgenic mutagenesis assay as a tool to evaluate mutational levels during normal and aberrant B cell differentiation. PMID- 9341755 TI - Biologically active, alternatively processed interleukin-1 beta in psoriatic scales. AB - The aims of the present work were to elucidate the biochemical properties of interleukin-1 beta (IL-1 beta) in psoriatic scales to get information on the processing of epidermal IL-1 beta in psoriasis, and to elucidate whether the IL-1 beta in psoriatic scales possesses biological activity. By means of ion exchange chromatography, IL-1 beta in extracts of psoriatic scales was purified to a stage where it could be analyzed with electrophoretic methods and immunoblotting. Compared to mature recombinant human IL-1 beta (Ala 117 IL-1 beta), IL-1 beta in psoriatic scales had a slightly higher apparent molecular mass and a more acidic isoelectric point, as revealed by two-dimensional electrophoresis under denaturing conditions. Isoelectric focusing under non-denaturing conditions of IL 1 beta partially purified from psoriatic scales, or from non-inflamed plantar stratum corneum (Nylander Lundqvist, E., Back, O. and Egelrud, T., J. Immunol. 1996. 157: 1699), and of mature IL-1 beta, followed by immunoblotting with IL-1 beta-specific antibodies, showed that psoriatic scales contained two components with IL-1 beta-like immunoreactivity which were isoelectric at pH 6.1 and 6.3, respectively. These components could also be detected in extracts of plantar stratum corneum, which also contained small amounts of an IL-1 beta-like component isoelectric at pH 6.9. Mature IL-1 beta was isoelectric at pH 6.9. No IL-1 beta-like biological activity could be detected in crude extracts of psoriatic scales. These extracts also contained high amounts of IL-1 receptor antagonist. Partially purified preparations of IL-1 beta from psoriatic scales, in which an apparently total separation of IL-1 beta and IL-1 receptor antagonist had been achieved, could induce expression of E-selectin in human umbilical vein endothelial cells. This activity was inhibited by antibodies specific for IL-1 beta, but not by antibodies specific for IL-1 alpha. It is concluded that psoriatic scales contain biologically active IL-1 beta, which has been processed by a mechanism which may be similar to that present in non-inflamed plantar stratum corneum, and which does not involve IL-1 beta converting enzyme. PMID- 9341756 TI - Induction of endothelial cell surface adhesion molecules by tumor necrosis factor is blocked by protein tyrosine phosphatase inhibitors: role of the nuclear transcription factor NF-kappa B. AB - Recent studies from our laboratory have indicated that protein tyrosine phosphatase (PTPase) inhibitors can down-modulate the tumor necrosis factor (TNF) mediated activation of the nuclear transcription factor NF-kappa B in ML-1a, a monocytic cell line (Singh and Aggarwal, J. Biol. Chem. 1995: 270: 10631). Since TNF is one of the major inducers of various adhesion molecules in human endothelial cells and their expression is known to require the activation of NF kappa B, we examined the effect of PTPase inhibitors on the TNF-mediated induction of intracellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1 and endothelial leukocyte adhesion molecule (ELAM)-1. Like ML 1a, human dermal microvessel endothelial cells (MVEC) treated with TNF rapidly activated (within 30 min) NF-kappa B; this effect was completely abolished by co treatment with phenylarsine oxide (PAO), a specific inhibitor of PTPase. The induction of ICAM-1, VCAM-1, and ELAM-1 by TNF in MVEC occurred within 6 h and was also completely down-regulated by PAO in a dose-dependent manner. PAO was found to be effective even when added 3 h after TNF, suggesting a rapid mode of action of this inhibitor. Besides PAO, other inhibitors of PTPase, including pervanadate and diamide, also blocked TNF-dependent NF-kappa B activation and induction of all the three adhesion proteins. Consistent with these results, the attachment of monocytes to MVEC was also blocked by the PTPase inhibitors. Thus, overall, our results demonstrate that a PTPase is involved either directly or indirectly in the pathway leading to the induction of endothelial cell adhesion molecules by TNF. Because of their role in cell adhesion, PTPase may provide a novel target of drug development for treatment of inflammation, atherogenesis, and tumor metastasis. PMID- 9341757 TI - The X-linked immunodeficiency defect in the mouse is corrected by expression of human Bruton's tyrosine kinase from a yeast artificial chromosome transgene. AB - Mutations in the gene for Bruton's tyrosine kinase result in the B cell differentiation defects X-linked agammaglobulinemia in man and X-linked immunodeficiency in mice. Here we describe the generation of two yeast artificial chromosome (YAC)-transgenic mouse strains in which high-level expression of human Btk is provided by endogenous regulatory cis-acting elements that are present on a 340-kb transgene, Yc340-hBtk. The expression pattern of the transgenic human Btk was found to parallel that of the endogenous murine gene. When the Yc340-hBtk transgenic mice were mated onto a Btk-deficient background, the xid B cell defects were fully corrected: conventional and CD5+ B-1 B cells were present in normal numbers, serum IgM and IgG3 levels as well as responses to T cell independent type II antigens were in the normal ranges. In vivo competition experiments in Btk+/- female mice demonstrated that in the conventional B cell population the Yc340-hBtk transgene could fully compensate the absence of expression of endogenous murine Btk. We conclude that in the YAC-transgenic mice Btk is appropriately expressed in the context of native regulatory sequences. PMID- 9341758 TI - Interleukin-2 receptor beta and gamma chain dysregulation during the inhibition of CD4 T cell activation by human immunodeficiency virus-1 gp120. AB - We have observed that CD4 T lymphocytes from human immunodeficiency virus (HIV) infected patients marginally express interleukin-2 receptor (IL-2R) beta and IL 2R gamma chains which are essential for IL-2 signal transduction. To analyze this observation further, we studied the influence of gp120 on the cell surface expression of IL-2R beta and IL-2R gamma by purified CD4 lymphocytes in vitro. Cross-linking of the T cell receptors of these lymphocytes initiates entry into the cell cycle as measured by CD69 and CD71 cell surface expression and [3H]thymidine incorporation. It also induces the cell surface expression of IL-2R beta and IL-2R gamma. We have shown that treatment of the CD4 T lymphocytes with HIV-1 gp120 before anti-CD3 stimulation impedes cell cycle progression as measured by reduced CD71 expression and inhibition of [3H]thymidine incorporation. Furthermore, cell surface expression of IL-2R beta and IL-2R gamma subunits, which from the functional intermediate-affinity IL-2R, are significantly inhibited. More importantly, addition of exogenous IL-2 does not restore the proliferation of the CD4 T cells treated with gp120, suggesting that cells are anergic and/or that the remaining IL-2R are not functional. This is the first study of IL-2R beta and IL-2R gamma dysregulation in the context of HIV infection and shows that CD4 is also involved in IL-2R expression. PMID- 9341759 TI - Peptide-induced T cell receptor down-regulation on naive T cells predicts agonist/partial agonist properties and strictly correlates with T cell activation. AB - Recent experiments defining T cell agonists, partial agonists and antagonists have suggested that the T cell can discriminate between subtle differences in interactions leading to T cell activation. To further understand the complexities of T cell activation, we have analyzed the requirements for the induction of a variety of effector functions using naive T cells and a variety of altered peptide ligands. Using a strong agonist peptide, massive T cell receptor (TCR) down-regulation correlated with a wide range of effector functions that were all induced above the same threshold peptide concentration. Interestingly, the kinetics of TCR down-regulation correlated with the concentration of the peptide, whereas the maximal degree of TCR down-regulation correlated with the induction of all monitored effector functions. A selected group of altered peptide ligands was also examined that were able to render target cells susceptible for lysis by effector cytotoxic T lymphocytes. The extent of TCR down-regulation induced by these peptides corresponded to the induction of a subset of effector functions. These studies have shown that the extent of TCR down-regulation defines the strength of TCR-mediated "signal 1" which correlates with the spectrum of effector functions activated within the T cell. Thus, activation of different T cell functions requires the triggering of distinct numbers of TCR. The different parameters that influence TCR down-regulation define important distinctions between our results and previously reported findings with T cell clones and may outline decisive parameters for the consequences of T cell activation in vivo. PMID- 9341760 TI - Enhanced sensitivity of P-glycoprotein-positive multidrug resistant tumor cells to complement-mediated lysis. AB - The interaction of KB-V1, a multidrug resistant (MDR) variant of the KB-3-1 human oral carcinoma, with human complement was investigated. KB-V1 cells were found to be more sensitive than KB-3-1 cells to complement-mediated lysis. Detailed analysis of the capacity of KB cells to activate human complement demonstrated that both C3b deposition and formation of the membrane attack complex (MAC) are higher on KB-V1 than on KB-3-1 cells. Furthermore, the MAC formed on KB-V1 cells, but not on KB-3-1 cells, was found to be resistant to trypsin treatment, i.e. more stably inserted into the plasma membrane. Immunofluorescence analysis by flow cytometry showed that KB-V1 cells express less decay-accelerating factor (DAF, CD55) than KB-3-1 cells. Two other complement regulatory proteins, membrane cofactor protein (MCP, CD46) and CD59 are expressed to a similar extent on both KB-V1 and KB-3-1 cells. Treatment of KB-V1 cells with neutralizing anti-P glycoprotein (P-gp) monoclonal antibodies reduced their sensitivity to complement. In addition, KB-V1 revertants which cease to express P-gp become more resistant to complement. These results indicate that multiple factors, such as reduced expression of DAF, enhanced deposition of C3b and increased binding and stability of the MAC may contribute to the increased complement sensitivity of KB V1 cells. It is suggested that P-gp is responsible for the complement-sensitive phenotype of KB-V1 cells. PMID- 9341761 TI - Expression and functions of the high-affinity IgE receptor on human platelets and megakaryocyte precursors. AB - Platelets can be activated by IgE and are therefore involved in IgE-mediated effector mechanisms against parasites and in allergic disorders. Here we show that, besides the low-affinity IgE receptor (Fc epsilon RII/CD23), platelets express the high-affinity receptor for IgE (Fc epsilon RI). Flow cytometry analysis revealed the existence of surface Fc epsilon RI on platelets with a large heterogeneity among individual donors, and a low proportion of platelets co expressing Fc epsilon RI and FC epsilon RII/CD23. Northern hybridization and reverse transcription polymerase chain reaction confirmed the presence of mRNA encoding the alpha, beta and gamma chains of Fc epsilon RI in platelets and in their megakaryocytic precursors. Cross-linking of Fc epsilon RI with monoclonal antibody (mAb) to alpha chain using either the whole molecule or F(ab')2 triggered platelet cytotoxicity for Schistosoma mansoni larvae. Anti-Fc epsilon RII/CD23 mAb significantly inhibited IgE- or Fc epsilon RI-mediated cytotoxicity, indicating down-regulatory effects of Fc epsilon RII/CD23 on Fc epsilon RI dependent functions. These results demonstrate functional properties for Fc epsilon RI on platelets and indicate unsuspected interactions between the low- and the high-affinity IgE receptors. PMID- 9341762 TI - Size-dependent effect of IgA on the IgA Fc receptor (CD89). AB - The IgA Fc receptor (FcR; CD89) is expressed on several types of cells of the myeloid cell lineage. We investigated whether different sizes of heat-aggregated IgA (aIgA) bind to CD89 and subsequently induce cellular activation. As a model we used the murine B cell line IIA1.6 transfected with CD89 or IIA1.6 cells transfected with CD89 as well as with the FcR gamma chain to study the binding of IgA to CD89. When these cells expressing CD89 were incubated with monomeric IgA, no significant binding of IgA to the cells was detectable by fluorescence activated cell sorter analysis; however, incubation of the cells with aggregated IgA resulted in 93 +/- 2% positive cells. Incubation of the cells with different sizes of IgA-containing aggregates revealed optimal binding with aggregates containing five to six molecules of IgA per aggregate. No difference was observed between the binding to CD89 of both IgA1- or IgA2-containing aggregates. Furthermore, the binding of aIgA was found to be CD89-specific, since the binding of IgA was completely inhibited by the CD89-specific monoclonal antibody My43 and no detectable binding occurred to the IIA1.6 parent cell line. Activation studies using interleukin-2 (IL-2) production as a marker, showed that the FcR gamma chain is necessary to induce cellular activation. Only cells transfected with both CD89 and the FcR gamma chain (CD89+/gamma +) enhance the IL-2 production 10 12-fold upon stimulation with aggregates of IgA. Furthermore, triggering of CD89 only results in increase of intracellular calcium concentration ([Ca2+]i) in cells co-expressing FcR gamma chain. Mutation of the tyrosine residues in the FcR gamma chain immunoreceptor tyrosine-based activation motif of the FcR gamma chain abolishes this increase in [Ca2+]i, indicating association and involvement of the FcR gamma chain in CD89-mediated signaling. PMID- 9341763 TI - The migratory behavior of murine CD4+ cells of memory phenotype. AB - Lymphocyte differentiation is connected with profound alterations in the migratory pattern of lymphocytes. Whereas naive cells predominantly recirculate through lymphoid tissues, activated lymphocytes acquire an increased preference for immigration into non-lymphoid tissues and a reduced capacity for recirculation via high endothelial venules (HEV). A variety of data had indicated that memory-related subpopulations of cells in man and sheep, classified by the low expression of the CD45RA isotype, also lack the capacity to recirculate via HEV. However, recent data in the rat called these results into question. We therefore analyzed the migration properties of murine CD4+ T cell subpopulations defined by several markers used to distinguish memory from naive CD4+ cells in mice, namely CD45RB, L-selectin and CD44. Our data clearly show that the majority of putative memory cells expressing either low levels of CD45RB, low levels of L selectin or high levels of CD44 display a strongly reduced capacity for direct entry into lymphoid tissues, including the spleen, from the blood stream. The accumulation in peripheral lymph nodes is further reduced by treatment with anti L-selectin antibody, which blocks their entry via HEV. This indicates that memory CD4+ T cells are not excluded from crossing lymph node HEV, and that the numbers of cells entering the node via this route exceed the numbers entering via the afferent lymph, at least in the absence of local inflammation. Concomitantly, a strongly enhanced localization of cells of the memory phenotype is observed in lung and liver as compared with naive cells. Trafficking to specific sites such as skin or gut mucosa is not a prominent feature of the total population of memory cells. The trafficking to lung and liver and an increased ability to bind to dendritic cells, demonstrable in in vitro adhesion assays, suggest a more sessile phenotype of most memory cells. With respect to these properties, memory cells have a surprizing similarity to fully activated lymphocytes. PMID- 9341764 TI - CD2-mediated signaling in T cells lacking the zeta-chain-specific immune receptor tyrosine-based activation (ITAM) motif. AB - T lymphocytes can be activated via the T cell receptor (TCR) or by triggering through a number of other surface structures, including the CD2 co-receptor molecule. Signaling through the CD2 molecule was shown previously to be dependent on the TCR-associated zeta-chain. Here, we show that CD2-induced activation also functions in T cells which express zeta-chains lacking a functional immune receptor tyrosine-based activation motif (ITAM). TCR-positive T cells that express only the transmembrane part of the zeta-chain protein and thus lack a functional zeta-derived ITAM readily produce interleukin (IL)-2 when cross-linked with CD2-specific monoclonal antibodies (mAb). TCR-negative T cell hybridomas expressing minimal receptors consisting of an extracellular CD25 and an intracellular zeta-chain-derived segment were effectively stimulated via CD2 specific mAb. For CD2-mediated co-stimulation of TCR-negative cells, two zeta chain-derived ITAM were sufficient to induce IL-2 when the CD2 molecules were co cross-linked with the chimeric CD25-zeta molecules. Taken together, our results show that CD2-induced signaling does not necessarily employ the zeta-chain in TCR positive cells and that CD2-dependent co-stimulation in TCR-negative cells can be mediated via two functional zeta-chain-derived ITAM. PMID- 9341765 TI - Opposite role for interleukin-4 and interferon-gamma on CD30 and lymphocyte activation gene-3 (LAG-3) expression by activated naive T cells. AB - Polarized human type 1 and type 2 T helper cells not only produce different sets of cytokines, but they also preferentially express certain activation markers, such as lymphocyte activation gene-3 (LAG-3) and CD30, respectively. In this study we have examined the LAG-3 and CD30 expression in relation to the lineage commitment of human naive CD4+ T cells, as assessed at the single-cell level of committed T cells. Purified CD45RA+ umbilical cord blood T lymphocytes were activated with phytohemagglutinin and interleukin (IL)-2 in the absence or presence of interleukin IL-4 or IL-12 and assessed for CD30 and LAG-3 expression, as well as for intracellular cytokine synthesis. Significant numbers of CD30+ cells were only found in CD4+ and CD8+ T lymphocytes of cultures primed with IL 4, which developed into cells able to produce IL-4 and IL-13 in addition to interferon (IFN)-gamma. By contrast, LAG-3 expression was strongly up-regulated in CD4+ and CD8+ T cells from cultures primed with IL-12, which developed into high numbers of IFN-gamma producers. The addition of a neutralizing anti-IFN gamma antibody to IL-12-primed CD4+ T cell cultures virtually abolished the development of LAG-3-expressing CD4+ T cells. Taken together, these data suggest that CD30 expression is dependent on the presence of IL-4, whereas LAG-3 expression is dependent on the production of IFN-gamma during the lineage commitment of human naive T cells. PMID- 9341766 TI - T cell repertoire in patients with multiple myeloma and monoclonal gammopathy of undetermined significance: clonal CD8+ T cell expansions are found preferentially in patients with a low tumor burden. AB - The T cell receptor (TCR) variable (V) gene repertoire was analyzed in patients with monoclonal gammopathy of undetermined significance (MGUS) (n = 17), multiple myeloma (MM) stage I (n = 16), MM stages II/III (n = 31) and age-matched controls (n = 27) by immunofluorescence and flow cytometry using a panel of mouse monoclonal antibodies (mAb) (n = 10) against TCR V alpha and V beta gene products. T cell expansion was defined as a value > or = thrice the normal median value for each respective TCR V mAb. Fifty-three percent of all patients displayed CD8+ expansion(s) as compared to 30% of age-matched controls (p < 0.001). Within the CD4 subset, 18% of the patients displayed T cell expansion(s) in comparison to 11% of the controls (not significant). Interestingly, the CD8+ expansion(s) were more frequently noted in patients with a low tumor burden (MGUS/MMI) (73%) as compared to those with advanced disease (MM II/III) (32% and control donors (30%) (p < 0.01). Likewise, multiple CD8+ expansions (two or more) were more common in MGUS/MM I patients than in MM II/III and controls (p < 0.01). The T cell expansions were stable over time in patients with a stable disease. A high degree of clonality of the expansions was detected by TCR CDR3 fragment length analysis, determination of J beta gene usage and nucleotide sequencing. The frequent finding of oligoclonal CD8+ T cell expansions in patients with a low tumor mass, but not in patients with advanced disease justifies further work in order to identify the relevance of expanded CD8+ T cells. In one patient with T cell reactivity against the autologous myeloma idiotype, two expansions within the CD8 population (V beta 3 and V beta 5.2 respectively) displayed no reactivity against the idiotype. Instead, idiotype recognition was confined to a CD8 non expanded V beta 22+ T cell population, with a highly restricted TCR usage (CDR3 fragment length analysis). PMID- 9341767 TI - A filarial cysteine protease inhibitor down-regulates T cell proliferation and enhances interleukin-10 production. AB - Filarial nematodes are a cause of chronic debilitating diseases in the tropics. A hallmark of filariasis is the marked down-regulation and polarization of host immune responses, yet molecular constituents of parasites causing this state have remained undefined. We describe a 17-kDa antigen (Av17) of the rodent filarial parasite Acanthocheilonema viteae, which shows amino acid homologies to cystatin C, a major cysteine protease inhibitor belonging to family 2 of the cystatin superfamily. Av17 is released by filariae in vitro. Exported molecules of A. viteae worms are shown to markedly suppress mitogen-induced T cell proliferation of mice and jirds. Av17 accounts for 45.5% of this suppressive activity in the murine system. Recombinant Av17 (rAv17), expressed in Escherichia coli, exhibits biological activity as a cysteine protease inhibitor and was used to examine the immunomodulatory effects, rAv17 induces down-regulation of murine T cell responses to mitogens, to T cell receptor cross-linking by anti-CD3 antibodies and to specific antigens, and at the same time up-regulation of interleukin-10. Hence, this filarial cystatin is a likely effector molecule of immunomodulation and a potential target for antifilarial intervention. PMID- 9341768 TI - Evidence for a CD4-associated calcium influx independent of the phosphoinositide transduction pathway in human T cells. AB - We recently showed, using human Jurkat T cell variants lacking the T cell receptor (TCR)/CD3 complex, that the lectin jacalin is able to trigger intracellular calcium increase provided that CD4 is expressed on the cell surface. Involvement of the CD4 molecule in jacalin-induced biological effects was furthermore demonstrated in differentiated U937 myelomonocytic cells expressing or not expressing CD4, and is confirmed here in human CD4-transfected mouse thymoma cells. In the present paper, we analyze the CD4-associated calcium response triggered by jacalin independently of the TCR/CD3 complex. We show that the observed calcium rise results from a direct long-lasting calcium influx from the outside without release of calcium from intracellular stores. We demonstrate that it is independent of the phosphoinositide phospholipase C transduction pathway. Moreover, we show that this peculiar calcium response can be blocked by protein tyrosine kinase inhibitors (herbimycin and genistein) giving evidence of the involvement of a protein tyrosine kinase, the best candidate of which is the CD4-associated p56lck. Altogether, our results suggest that, independently of the TCR/CD3 complex, CD4 may be involved in the triggering of a calcium signal dependent on a protein tyrosine kinase and independent of the phosphoinositide transduction pathway. PMID- 9341769 TI - Proteasome regulation of Fas ligand cytotoxicity. AB - The role of NF-kappa B in regulating FasL-mediated cytotoxicity was investigated by using lactacystin. Lactacystin is a microbial metabolite known to inhibit only the protease activity of the proteasome, which is required for NF-kappa B translocation. When activated by immobilized anti-CD3 monoclonal antibody, hybridoma T cells (5D5) degraded I kappa B beta, translocated NF-kappa B into the nucleus, transcribed immediate-early genes and the Fas ligand (FasL) gene, and expressed FasL-mediated cytotoxicity. Lactacystin strongly blocked I kappa B beta degradation and the translocation of NF-kappa B (p50/RelA heterodimer), but had little effect on the expression of the transcription factors, Oct-1 and AP-1. Moreover, lactacystin did not inhibit the nuclear translocation of NF-ATp whereas cyclosporin A inhibited the translocation of both NF-kappa B and NF-ATp. The expression of c-myc and nur77, two immediate-early genes implicated in FasL gene activation, was blocked by lactacystin. Subsequently, the expression of FasL gene and FasL-mediated cytotoxicity was inhibited. LLnL, a well-known peptide aldehyde which inhibits the protease activities of the proteasome and cysteine proteases, also inhibited NF-kappa B translocation and FasL-mediated cytotoxicity. However, these events were not inhibited by the highly specific cysteine protease inhibitor E64. These observations provide further evidence that FasL cytotoxicity is regulated by the proteasome. Furthermore, lactacystin must be added early in order to efficiently inhibit the induction of FasL cytotoxicity, indicating that the early events are critical for FasL gene activation. Our study integrates the proteasome-dependent I kappa B degradation and NF-kappa B translocation into a T cell activation cascade which results in FasL gene activation and the expression of FasL-mediated cytotoxicity. PMID- 9341770 TI - Double-positive T cell receptor(high) thymocytes are resistant to peptide/major histocompatibility complex ligand-induced negative selection. AB - To investigate negative selection events during intrathymic ontogeny, we established T cell receptor (TCR)-transgenic mice [N15tg/RAG-2-/- (H-2b)] expressing a single TCR specific for vesicular stomatitis virus nuclear octapeptide N52-59 (VSV8) in the context of the major histocompatibility complex (MHC) class I molecule, K(b). Administration of VSV8 in vivo induced apoptosis in less than 4 h, deleting the majority of immature double-positive (DP) thymocytes by 24 h. In contrast, DP TCRhigh as well as single-positive (SP) thymocytes were refractory to this death process. Moreover, DP TCRhigh cells differentiated into SP thymocytes in vitro and in vivo, maturing into functional cytotoxic T lymphocytes upon intrathymic transfer to beta RAG 2-/- recipients. Hence, negative selection processes involving MHC-bound peptide ligands are operative only prior to the late DP thymocyte stage in this MHC class I-restricted TCR transgene system. PMID- 9341771 TI - Mouse CD23 regulates monocyte activation through an interaction with the adhesion molecule CD11b/CD18. AB - CD23 is expressed on a variety of hemopoietic cells. Recently, we have reported that blocking CD23 interactions in a murine model of arthritis resulted in a marked improvement of disease severity. Here, we demonstrate that CD11b, the alpha chain of the beta 2 integrin adhesion molecule complex CD11b/CD18 expressed on monocytes interacts with CD23. Using a recombinant fusion protein (ZZ-CD23), murine CD23 was shown to bind to peritoneal macrophages and peripheral blood cells isolated from mice as well as the murine macrophage cell line, RAW. The interactions between mouse ZZ-CD23 and CD11b/CD18-expressing cells were significantly inhibited by anti-CD11b monoclonal antibodies. A functional consequence was then demonstrated by inducing an up-regulation of interleukin-6 (IL-6) production following ZZ-CD23 incubation with monocytes. The addition of Fab fragments generated from the monoclonal antibody CD11b impaired this cytokine production by 50%. Interestingly, a positive autocrine loop was identified as IL 6 was shown to increase CD23 binding to macrophages. These results demonstrate that similar to findings using human cells, murine CD23 binds to the surface adhesion molecule, CD11b, and these interactions regulate biological activities of murine myeloid cells. PMID- 9341772 TI - Human mast cells express functional TrkA and are a source of nerve growth factor. AB - Mast cells are the principal effector cells in IgE-dependent hypersensitivity reactions. Despite reports that rodent mast cells proliferate in the presence of nerve growth factor (NGF), human mast cells reportedly do not respond to this factor. To determine if human mast cells express the NGF receptors, TrkA tyrosine receptor and the low affinity NGF receptor (LNGFR), we first analyzed the mRNA expression by RT-PCR of TrkA and LNGFR in a human mast cell line (HMC-1) and in human mast cells cultured in the presence of stem cell factor. Both HMC-1 and cultured human mast cells were found to express TrkA but not LNGFR. TrkA protein was demonstrated by Western blot analysis of HMC-1 lysates. Using flow cytometric analysis and mast cell tryptase as a mast cell marker, both HMC-1 cells and cultured human mast cells were shown to coexpress tryptase and TrkA. Treatment of mast cells with NGF resulted in phosphorylation of TrkA on tyrosine residues as detected by immunoblotting with an antiphosphotyrosine antibody. Furthermore, NGF induced the immediate early gene c-fos in HMC-1 cells. HMC-1 cells and cultured human mast cells were also found to express NGF mRNA, and conditioned medium from HMC-1 cells stimulated neurite outgrowth from chicken embryonic sensory ganglia in culture. This effect was blocked by anti-NGF. Thus, mast cells express functional TrkA and synthesize NGF, suggesting a mechanism by which NGF may act as an autocrine factor for human mast cells, and by which mast cells and nerves may interact. PMID- 9341773 TI - Nephritogenic T cells use granzyme C as a cytotoxic mediator. AB - Cytoplasmic granules of cytotoxic T lymphocytes contain several proteins that may be involved in cell-mediated cytotoxicity. We have previously described nephritogenic T cell clones that are cytotoxic to cultured renal proximal tubular epithelial cells (MCT). One of these clones, M52.34.1, expresses perforin, a cytotoxic mediator. We investigated the expression of other granule-associated proteases of M52.34.1. Granzymes A and B have been extensively studied in T cell mediated cytotoxicity, and associated with tissue destruction in models of transplantation. However, the activity of other granzymes has not been as extensively investigated. We focused our studies on granzyme C. Northern blots showed very high levels of granzymes B and C mRNA expression in M52.34.1 cells 3 days following T cell activation. There was no expression of granzyme A mRNA. An antisense oligonucleotide made from the 5'-upstream region of the murine granzyme C exon 1 inhibited granzyme C mRNA expression in M52.34.1 when added at a concentration of 50 microM to the culture medium for 2 days. There was no inhibition of granzyme C mRNA expression with the sense oligonucleotide. The granzyme C antisense oligonucleotide inhibited M52.34.1 cytotoxicity to MCT at effector:target ratios of 20:1 and 40:1. M52.34.1 cells mediate inflammatory interstitial nephritis following adoptive transfer. If T cells were resuspended in 200 microM of the antisense oligonucleotide prior to subcapsular transfer, the recipient kidneys showed markedly diminished tubular cell destruction, suggesting that granzyme C can also be an important mediator of cytotoxicity in vivo. PMID- 9341774 TI - Antigen-specific CD8+ T cell subset distribution in lymph nodes draining the site of herpes simplex virus infection. AB - Inoculation with replicating virus leads to an increase in T cell numbers within lymph nodes that drain the site of infection. This increase has been associated with a nonspecific proliferation of bystander cells, with only a minority thought to be directed to the infectious agent. Such an assumption is largely based on precursor cytotoxic T lymphocyte (CTL) estimations using limiting dilution analysis. Recently, studies using more advanced molecular approaches have suggested that such functionally derived precursor frequencies considerably underestimate the proportion of T cells specific for the antigen under investigation. We have defined T cell receptor sequences characteristic of CTL populations directed to a dominant determinant of the herpes simplex virus (HSV) glycoprotein B (gB). In this investigation, we used this receptor signature as a probe to directly monitor changes occurring within lymph nodes draining the sites of active infection with HSV. We found that although lymph node CD8+ T cell numbers increase as a consequence of HSV infection, the majority of these cells are small resting cells that are not enriched for gB-specific receptors. In contrast, a significant proportion of activated T cells are highly enriched for CTL bearing gB-specific receptors. Our results are therefore consistent with a nonspecific migration of CTL precursors into the lymph nodes draining the site of infection, followed by the activation and proliferation of the antigen-specific subset that normally makes up a small proportion of the naive T cell repertoire. PMID- 9341775 TI - Blood- and skin-derived monocytes/macrophages respond to C3a but not to C3a(desArg) with a transient release of calcium via a pertussis toxin-sensitive signal transduction pathway. AB - Controversial results have been published in the past regarding the functional reactivity of monocytes (Mo) and macrophages (M phi) to the anaphylatoxin C3a and its degradation product C3a(desArg). In this study we performed binding and calcium mobilization experiments with recombinant human C3a (rC3a) and rC3a(desArg). Blood Mo displayed non-inhibitable binding of FITC-labeled rC3a (rC3aFITC) but responded to rC3a with a transient release of the intracellular calcium concentration ([Ca2+]i), whereas rC3a(desArg) was completely inactive. In contrast, binding of rC3aFITC to eosinophilic granulocytes and the mast cell line HMC-1 which have been shown previously to express C3a binding sites could be blocked by a monoclonal anti-C3a antibody. The rC3a-induced [Ca2+]i release in blood Mo was pertussis toxin (PTX)-sensitive suggesting the involvement of G proteins in the signal transduction pathway. Skin-derived Mo/M phi reacted similarly to blood Mo as no specific binding of rC3aFITC to these cells could be demonstrated, whereas an intracellular release of calcium ions in response to the anaphylatoxin was observed. Homologous desensitization to rC3a but not heterologous desensitization to rC5a was detected in further experiments. The functional effect of C3a, but not the unspecific binding of rC3aFITC to blood Mo and skin-derived Mo/M phi could be blocked by the monoclonal anti-C3a antibody. These results suggest the expression of the recently cloned G-protein-coupled receptor for C3a on human blood Mo and skin-derived Mo/M phi. However, the total number of specific C3a binding sites on these cells is distinctly lower as compared to eosinophilic granulocytes and cells of the mast cell line HMC-1. The small number of C3a receptors on Mo/M phi may be masked by a pronounced non inhibitable binding of rC3aFITC. This binding, however, may contribute to the recently described biological effects of C3a(desArg) on Mo. PMID- 9341776 TI - Antagonism of cytotoxic T lymphocyte-mediated lysis by natural HIV-1 altered peptide ligands requires simultaneous presentation of agonist and antagonist peptides. AB - Mutations in human immunodeficiency virus (HIV) cluster in cytotoxic T lymphocyte (CTL) epitopes (Phillips, R. E. et al., Nature 1991. 354: 453) and are subject to immune-mediated positive selection (Price, D. A. et al., Proc. Natl. Acad. Sci. USA 1997. 94: 1890). We studied the effects of naturally occurring mutations in the HIV-1 p17 Gag HLA A2 restricted epitope SLYNTVATL on recognition by anti-HIV CTL. Most of these naturally occurring mutants escaped killing by one CTL line and the majority acted as CTL antagonists. We also investigated whether CTL exposed to a strict antagonist peptide restricted by HLA A2 were unresponsive when exposed to targets presenting the wild-type sequence. The results show that antagonism of anti-HIV CTL killing requires the simultaneous presence of agonist and antagonist peptide. We found no evidence that CTL exposed to an antagonist received a functionally negative signal since these CTL retained an unimpaired capacity to lyse targets bearing wild-type peptide. PMID- 9341777 TI - Deviation of pancreas-infiltrating cells to Th2 by interleukin-12 antagonist administration inhibits autoimmune diabetes. AB - Nonobese diabetic (NOD) mice develop spontaneous insulin-dependent diabetes mellitus (IDDM), and the pancreas-infiltrating T cells invariably show a Th1 phenotype. We demonstrated here that the interleukin (IL)-12 antagonist (p40)2 can deviate the default Th1 development of naive T cell receptor (TCR)-transgenic CD4+ cells to the Th2 pathway in vitro. Although (p40)2 does not modify the cytokine profile of polarized Th1 cells, it prevents further recruitment of CD4- cells into the Th1 subset. To study the involvement of Th1 and Th2 cells in the initiation and progression of IDDM, we targeted endogenous IL-12 by administration of (p40)2 in NOD mice. (p40)2 administration to NOD mice inhibits interferon-gamma but not IL-10 production in response to lipopolysaccharide (LPS) or to the putative autoantigen IA-2. Serum immunoglobulin isotypes determined after (p40)2 treatment indicate an increase in Th2 and a decrease in Th1 helper activity. Administration of (p40)2 from 3 weeks of age onwards, before the onset of insulitis, results in the deviation of pancreas-infiltrating CD4+ but not CD8+ cells to the Th2 phenotype as well as in the reduction of spontaneous and cyclophosphamide-accelerated IDDM. After treating NOD mice with (p40)2 from 9 weeks of age, when insulitis is well established, few Th2 and a reduced percentage of Th1 cells are found in the pancreas. This is associated with a slightly decreased incidence of spontaneous IDDM, but no protection from cyclophosphamide-accelerated IDDM. In conclusion, deviation of pancreas infiltrating CD4+ cells to Th2 is associated with protection from IDDM. However, targeting IL-12 after the onset of insulitis, when the pancreas contains polarized Th1 cells, is not sufficient to induce an effective immune deviation able to significantly modify the course of disease. PMID- 9341778 TI - CpG-containing synthetic oligonucleotides promote B and cytotoxic T cell responses to protein antigen: a new class of vaccine adjuvants. AB - Foreign DNA has been shown to impinge on immune cell function by an as yet unidentified mechanism. We and others have demonstrated that single-stranded (ss) DNA containing the motif CpG flanked by two 5' purines and two 3' pyrimidines are mitogenic for B cells and activate macrophages to release tumor necrosis factor alpha, interferon-gamma, interleukin (IL)-6 or IL-12. Because of these pro inflammatory responses we investigated if ssDNA would serve as a potential vaccine adjuvant. Here we show that CpG-containing oligonucleotides represent a powerful adjuvant for both humoral and cellular immune responses. When ssDNA was incorporated into inocula, specific antibody titers of the IgG2 isotype were enhanced by greater than 100-fold. Primary cytotoxic T lymphocyte responses generated to either unprocessed protein antigen or major histocompatibility complex class I-restricted peptide were exceedingly strong. Evidence is also provided that oligomers directly influenced T cell receptor-triggered T cell proliferation. Thus ssDNA oligomers may serve as inexpensive and safe vaccine adjuvants and, in addition, differential effects due to sequence may allow for directed responses. PMID- 9341779 TI - Phenotypic and functional analysis of CD4+ NKRP1A+ human T lymphocytes. Direct evidence that the NKRP1A molecule is involved in transendothelial migration. AB - In this report, we show that among human CD4+ T lymphocytes 5-20% express the C type lectin molecule NKRP1A. This lymphocyte subset displays a slightly more limited T cell receptor V beta repertoire than the CD4+ NKRP1A- counterpart. CD4+ NKRP1A+ T lymphocytes are characterized by a high expression of beta 1 and beta 2 integrins, thus representing a T lymphocyte subset that can possibly adhere and migrate through vascular endothelium. Indeed, resting CD4+ NKRP1A+ lymphocytes, differently from the CD4+ NKRP1A- subset, migrated across endothelial cell monolayers in a Transwell chamber system. Pretreatment of CD4+ NKRP1A+ T lymphocytes with an anti-NKRP1A monoclonal antibody (mAb) strongly reduced transendothelial migration, suggesting the involvement of the NKRP1A molecule in the transmigration process. Furthermore, cells of the NKRP1A- Jurkat CD4+ T cell line stably transfected with NKRP1A cDNA migrated more rapidly and efficiently than either untransfected or mock-transfected Jurkat cells. Finally, mAb-mediated cross-linking of NKRP1A molecules in CD4+ T lymphocytes induced the up-regulation of the lymphocyte function-associated antigen 1 Mg(2+)-binding site as well as beta 1 and beta 2 integrin chains. Altogether, these findings suggest that the NKRP1A molecule is involved in transendothelial migration of resting CD4+ T lymphocytes. PMID- 9341781 TI - An Fc gamma receptor I (CD64)-negative subpopulation of human peripheral blood monocytes is resistant to killing by antigen-activated CD4-positive cytotoxic T cells. AB - It has been demonstrated that in monocyte/T cell co-cultures activated with recall antigens, cytotoxic T cells were generated which are able to reduce the number of antigen-presenting monocytes. In previous studies we could show that a minor subset of monocytes, the Fc gamma receptor I-negative (CD64-) monocytes, exhibits significantly higher antigen-presenting capacity than the main population of monocytes (> 90%) which are Fc gamma receptor I-positive (CD64+). Therefore, we addressed the question whether they are also differentially susceptible to T cell-mediated killing. In the present study we demonstrate that the CD64- monocyte subset is more resistant to killing by antigen-activated T cells than CD64+ monocytes, as indicated by a higher viability and recovery of CD64- monocytes. This mechanism involves CD95 (Fas) antigen, since monocyte death in co-cultures with antigen-activated T cells could be partially reduced by blocking anti-Fas monoclonal antibodies (mAb). In agreement with this finding, although CD95 antigen was expressed on CD64+ and CD64- monocytes at comparable levels, killing of CD64- monocytes by activating anti-Fas mAb was lower than of CD64+ monocytes. PMID- 9341780 TI - Glucocorticoids inhibit human antigen-specific and enhance total IgE and IgG4 production due to differential effects on T and B cells in vitro. AB - Although anti-inflammatory properties of glucocorticoids (GC) are well documented, their activity in allergic diseases is still controversial. Recently, it has been reported that GC can increase, both in vivo and in vitro, the polyclonal production of total IgE. In this study we investigated the effects of GC on the antigen (Ag)-specific IgE response in a human in vitro system with peripheral blood mononuclear cells or B cells of bee venom-sensitized individuals that allows the production of bee venom phospholipase A2 (PLA)-specific IgE and IgG4 antibodies (Ab). PLA-specific Ab were induced by simultaneously activating T cells and B cells specifically with allergen and polyclonally with anti-CD2 and soluble CD40 ligand (sCD40L) in the presence of interleukin (IL)-4. Indeed, dexamethasone and prednisolone enhanced the formation of total IgE and IgG4 in PBMC, while the production of PLA-specific IgE and IgG4 Ab was selectively inhibited in a dose-dependent manner. The suppressive effect of GC was mediated during Ag-specific stimulation and T cell-B cell interaction. This was due to GC suppressing specific T cell proliferation and cytokine production, whereas neither allergen-specific nor total IgE and IgG4 production by sCD40L/IL-4 stimulated pure B cells was affected. In contrast to GC, cyclosporine A inhibited both total and PLA-specific IgE and IgG4 secretion in peripheral blood mononuclear cells and B cell cultures. Further experiments showed that increase in nonspecific total isotype response resulted from inhibition of IL-4 uptake by cells other than B cells and sufficient availability of IL-4 to B cells for isotype switch and synthesis. Furthermore, demonstration of opposite regulatory effects of GC on specific and total isotype formation in vitro, including the inhibition of allergy-relevant Ag-specific IgE response, may contribute to a better understanding of apparently controversial observations, and explain why most allergic patients benefit from GC therapy. PMID- 9341782 TI - Marginal zone B cells exhibit unique activation, proliferative and immunoglobulin secretory responses. AB - Mouse splenic B cells can be separated based on their distinctive expression of cell surface antigens. Marginal zone (MZ) B cells are CD38high CD21high CD23low/ , while follicular (FO) B cells are CD21int CD23int and newly formed (NF) B cells are CD21dim/- CD23-. Exploiting these phenotypic distinctions, we isolated the three B cell subsets and assessed their other phenotypic differences and functional capabilities in vitro. FO B cells proliferate more than the other B cell subsets in response to either IgM or CD38 cross-linking. MZ B cells proliferate better than FO B cells when stimulated with lipopolysaccharide (LPS), sub-optimal levels of LPS and CD38 cross-linking or CD40 ligation. When NF, FO and MZ B cells were stimulated with either LPS or LPS and interleukin-4, MZ B cells secreted more IgM and IgG3 than the other two subsets. Similarly, calcium fluxes measured within MZ B cells are larger in amplitude and more sustained after B cell receptor cross-linking than those found in the other two subsets. Collectively, these results indicate that CD38high CD21high CD23low/- MZ B cells are specially suited to play a unique role in response to antigens delivered to the MZ area. PMID- 9341783 TI - Genetic modification of a carcinoma with the IL-4 gene increases the influx of dendritic cells relative to other cytokines. AB - Tumor cells genetically modified with certain cytokine genes gain immunogenic properties that allow the development of systemic anti-tumor immunity. Whether different cytokines may influence infiltration of transduced tumors by dendritic cells (DC) has not been investigated. Therefore, we analyzed the C26 murine colon carcinoma genetically modified to release interleukin (IL)-2, IL-4, IL-12, granulocyte colony-stimulating-factor (CSF) or granulocyte-macrophage (GM)-CSF for immunostaining with the monoclonal antibody NDLC145 recognizing the DEC205 determinant which, on tumor sections, is virtually restricted to DC. Infiltrating leukocytes were also characterized for expression of co-stimulatory molecules like CD54, CD86 and major histocompatibility complex class II. The intratumoral DC content was dependent on the type of transduced cytokines with C26/IL-4 being the most abundant in DEC205+ cells. The effect of IL-4 in recruiting DC did not depend on the type of tumor since it was confirmed in the TSA mammary carcinoma. In comparison with C26/GM-CSF, C26/IL-4 had more B7.2+ cells but less Ia+ cells. Furthermore, the hypertrophic skin overlaying tumors producing GM-CSF showed numerous Langerhans cells stained by NDLC145 and the draining lymph nodes showed abundance and paucity of DC in C26/GM-CSF and C26/IL-4, respectively. When injected into the ear pinna, C26/GM-CSF stimulated, whereas C26/IL-4 inhibited DC mediated priming of delayed-type hypersensitivity reaction by 2,4-dinitro-1 fluorobenzene. These findings prove that transduced cytokines differently influence DC recruitment at the tumor site and DC function in nearby tissues. Along with the other leukocytes and their secondary produced cytokines, DC create an environment in which T cells can be differently modulated. Such a phenomenon may have implications on genetic modification of tumor cells to be used as cancer vaccine. PMID- 9341784 TI - In vitro responses of human CD45R0brightRA- and CD45R0-RAbright T cell subsets and their relationship to memory and naive T cells. AB - The cellular basis of immunological memory, particularly with respect to T cells is not understood. In humans, monoclonal antibodies to CD45 have been used to identify memory (CD45R0) and naive (CD45RA) T cells. However, this identification has been called into question by various studies which suggest that high molecular weight CD45 isoforms may be re-expressed by previously activated cells. In the present study, using cultures which supported responses of naive T cells, we examined the responses of purified CD45R0brightRA- or CD45R0(-)-RAbright T cell subsets. The former subset was found to respond preferentially to recall antigens with minimal responses apparent to neo-(or non-recall)-antigens. The inverse pattern was found for CD45R0-RAbright T cells, which converted to CD45R0brightRA- after stimulation with a neo-antigen. Moreover, the two populations of T cells exhibited distinct response kinetics with a faster response evident from the CD45R0brightRA- T cells compared to the CD45R0-RAbright subset. The poor responses of CD45R0-RAbright T cells to recall antigens compared to neo-antigens suggests that this putative naive population is specifically depleted of reactive T cells following an encounter with antigen. We propose that T cell priming results in the stimulation of many CD45R0-RAbright T cells with various T cell receptor specificities from which memory T cells are selected for survival. If re-expression of higher molecular weight isoforms does occur in humans in vivo, our results suggest that R0 expression would be retained (CD45R0+RA+). Alternatively, if primed CD45R0-RAbright T cells exist, they are not prevalent in peripheral blood and thus may be sequestered within lymphoid tissues. Our data support the view that in human peripheral blood, CD45R0bright and CD45RAbright expression identify memory and naive CD4+ T cells, respectively. PMID- 9341785 TI - Priming of cytotoxic T lymphocytes by five heat-aggregated antigens in vivo: conditions, efficiency, and relation to antibody responses. AB - Mice were immunized i.p. with soluble or heat-denatured protein antigens [ovalbumin, beta-galactosidase, or recombinant E7 protein of human papilloma virus type 16 (HBV)]. Heat-denatured (100 degrees C) preparations of these proteins were able to induce cytotoxic T lymphocytes (CTL) that recognize cells expressing the respective genes, whereas native protein was either inefficient or required up to 30-fold higher doses. If the heat-treated proteins were separated into aggregated and soluble fractions by ultracentrifugation, only the aggregated fractions were able to induce specific CTL; this is probably because of the easier access to one of the major histocompatibility complex class I loading pathways for exogenous antigen. Addition of the adjuvant aluminium hydroxide (alum) to aggregated proteins abolished their ability to induce CTL; thus, a condition leading to a strong antibody response appeared to inhibit CTL induction. Interestingly, immunization with heat-denatured ovalbumin plus alum increased the IgM/IgG1 ratio compared to immunization with native ovalbumin and alum. Immunization of B6 mice transgenic for an HLA-A2/H-2K(b) hybrid gene with heat-denatured, recombinant HPV 16-E7 protein induced D(b)-restricted CTL specific for the peptide 49-57 of E7, indicating that this epitope is immunodominant over any A2-restricted E7 epitope in these mice. A whole influenza virus preparation heated to 100 degrees C or even autoclaved was still able to induce virus-specific CTL and BALB/c spleen cells heated to 100 degrees C could still cross-prime minor H-specific CTL in B6 mice, although with lower efficiency than fresh spleen cells. Thus, aggregated proteins can be considered as components for future vaccines. PMID- 9341786 TI - Priming of helper T cell-dependent antibody responses by hemagglutinin-transgenic B cells. AB - Mice expressing the hemagglutinin (HA) gene of influenza virus PR8 (H1 subtype) under the control of kappa light chain promoter and enhancer have been generated. They express HA in and on B cells, and are tolerant to HA. In vitro, only lipopolysaccharide (LPS) blasts but not resting B cells of transgenic mice can stimulate HA-specific helper T cells of HA-specific alpha/beta T cell receptor (TCR)-transgenic mice. Transfer of HA-transgenic LPS blasts into syngeneic, non transgenic recipients primes HA-specific antibody responses. Resting, small HA transgenic B cells, which were purified by fluorescence-activated cell sorting, prime lower antibody responses. Host B cells produce the HA-specific antibody response. The donor HA-transgenic B cells need to express major histocompatibility complex (MHC) class II molecules and need to be alive to induce the antibody response in the host. Most notably, the host antibody response never produces detectable levels of IgM, but only of switched IgG isotypes. Neither resting nor activated HA-transgenic B cells induce tolerance in antibody responses. These results suggest that HA-transgenic B cells, presenting both the intact antigen on the cell surface and peptides of the antigen on MHC class II, are effective inducers of helper T cell responses, and as judged by the Ig-isotype response pattern, which is mainly IgG1, of Th2 type. PMID- 9341787 TI - Presentation of a viral peptide assembled on the carbohydrate moieties of immunoglobulin does not require processing. AB - We have previously demonstrated that an immunodominant CD4 T cell epitope, HA110 120 of the hemagglutinin (HA) of the A/PR/8/34 influenza virus, enzymatically assembled on the carbohydrate moieties of self immunoglobulins (Ig) primed the precursors of peptide-specific T cells and induced efficient proliferation in vivo of naive lymphocytes from transgenic mice expressing the peptide-specific T cell receptor. Here, we show that an immuno-galacto-peptide construct, IgG-gal HA, does not require intracellular or extracellular processing to present the peptide to the specific T cells. The presentation occurs following the binding of the IgG-gal-HA construct to Fc gamma receptor on the surface of antigen presenting cells (APC), with concurrent interaction of the peptides to their neighboring major histocompatibility complex class II molecules. This mechanism of peptide presentation may harness the immune response in vivo by the engagement of APC with a low capacity of antigen processing, such as neonatal B cells. In addition, the enzymatic method of assembling various aminated compounds on the sugar moieties of Ig may offer novel perspectives on immuno-targeting of antagonist peptides, cytostatic drugs, and biologically active ligands of therapeutic use. PMID- 9341788 TI - The mannose receptor functions as a high capacity and broad specificity antigen receptor in human dendritic cells. AB - Dendritic cells, in contrast to B lymphocytes, must be able to efficiently internalize a diverse array of antigens for processing and loading onto major histocompatibility complex (MHC) class II molecules. Here we characterize the mannose receptor pathway in dendritic cells and show that mannose receptor mediated uptake of antigens results in a approximately 100-fold more efficient presentation to T cells, as compared to antigens internalized via fluid phase. Immunocytochemistry as well as subcellular fractionation revealed the localization of the mannose receptor and MHC class II molecules in distinct subcellular compartments. The mannose receptor thus functions in rapid internalization and concentration of a variety of glycosylated antigens that become available for processing and presentation. This may contribute to the unique capacity of dendritic cells to generate primary T cell responses against infectious agents. PMID- 9341789 TI - Mannose receptor-mediated uptake of antigens strongly enhances HLA class II restricted antigen presentation by cultured dendritic cells. AB - Dendritic cells (DC) efficiently take up antigens by macropinocytosis and mannose receptor-mediated endocytosis. Here we show that endocytosis of mannose receptor antigen complexes takes place via small coated vesicles, while non-mannosylated antigens were mainly present in larger vesicles. Shortly after internalization the mannose receptor and its ligand appeared in the larger vesicles. Within 10 min, the mannosylated and non-mannosylated antigens co-localized with typical markers for major histocompatibility complex class II-enriched compartments and lysosomes. In contrast, the mannose receptor appeared not to reach these compartments, suggesting that it releases its ligand in an earlier endosomal structure. Moreover, we demonstrate that mannosylation of protein antigen and peptides resulted in a 200-10,000-fold enhanced potency to stimulate HLA class II restricted peptide-specific T cell clones compared to non-mannosylated peptides. Our results indicate that mannosylation of antigen leads to selective targeting and subsequent superior presentation by DC which may be applicable in vaccine design. PMID- 9341790 TI - Monoclonal gammopathies in aging mu, kappa-transgenic mice: involvement of the B 1 cell lineage. AB - Monoclonal gammopathies (MG) are monoclonal proliferative disorders of B cells at the differentiation stage of Ig production. They can be detected in the serum, either as transient or as persistent homogenous immunoglobulin (H-Ig) components. The exact phenotype, localization, and cell lineage origin of the precursor cells of MG are unknown, but may be crucial for both the correct diagnosis and for timely efficient treatment of the malignant forms. We used for the first time transgenic (Tg) mice (Sp6; mu/kappa) to study the origin of MG. In the mu, kappa Tg mice a small proportion of B cells can still produce endogenous IgM. These cells are of B-1 cell origin. The MG in Tg mice showed a later onset and a lower frequency than those in littermate control mice, mainly due to a four times lower frequency of benign monoclonal gammopathy. The 10% of B-1 cells that were able to produce endogenous Ig led to the development of MG in a frequency that was half the number of MG found in normal littermates. None of the MG in Tg mice produced an Ig of the Tg origin and therefore it can be concluded that they originated from B-1 cells. PMID- 9341791 TI - TAP-translocated peptides specifically bind proteins in the endoplasmic reticulum, including gp96, protein disulfide isomerase and calreticulin. AB - The endoplasmic reticulum (ER) membrane-embedded transporter associated with antigen processing (TAP) associates with peptides in the cytosol and translocates these into the ER lumen. Here, MHC class I molecules bind a subset of these peptides and the remainder is either removed or degraded, or may be retained in the ER in association with other proteins. We have visualized peptide-binding proteins in the ER using radioactive peptides with a photoreactive group. Besides TAP, two proteins were identified as gp96 and protein disulfide isomerase (PDI). Calreticulin, previously found in complex with TAP, only binds glycosylated peptides. In addition, two as yet unidentified, ER luminal glycoproteins (gp120 and gp170) were visualized. The effects of peptide size and sequence on binding to the ER-resident proteins were studied by using partially degenerated peptides with photoreactive side chains. All identified proteins were able to bind peptides within the size range of peptides translocated by TAP, from 8 to more than 20 amino acids. Whereas PDI associated with all peptides tested, gp96 and gp120 showed a clear sequence preference for non-charged amino acids at positions 2 and 9 in 9mer peptides. Thus various ER proteins, other than the MHC class I heterodimer and TAP, are able to interact with peptides albeit with a different substrate selectivity. PMID- 9341792 TI - Mycobacterium bovis Bacillus Calmette Guerin infection prevents apoptosis of resting human monocytes. AB - Apoptosis plays an essential role in the development and homeostasis of multicellular organisms. Some infectious agents interfere with this programmed cell death to their own benefit. Here, we show that infection of resting human monocytes with Mycobacterium bovis Bacillus Calmette Guerin (BCG) increases monocyte viability by preventing them from undergoing apoptosis. Heat-killed BCG also prevented apoptosis, indicating that replication of BCG is not required to prevent cell death. Analysis of BCG-infected monocytes revealed an up-regulation of the A1 mRNA, whereas the bcl-2 mRNA was not up-regulated. Interestingly, preinfection with BCG renders the cells resistant to interleukin (IL)-10-induced apoptosis which may be one of the mechanisms mycobacteria use to modulate immune responses. BCG infection was also accompanied by an impairment of the capacity of monocytes to secrete IL-10 and by an induction of the capacity to secrete tumor necrosis factor-alpha, two cytokines known to induce and prevent human monocyte apoptosis, respectively. Since it has been reported that apoptosis is involved in killing of intracellular mycobacteria, the prevention of apoptosis may represent a strategy for mycobacterial survival in the infected host. PMID- 9341794 TI - Overexpression of the death-promoting gene bax-alpha sensitizes human BL-41 Burkitt lymphoma cells for surface IgM-mediated apoptosis. AB - Major regulators of programmed cell death, or apoptosis, are the members of the bcl-2 gene family. Recently, we reported that surface(s) IgM triggering of the human B lymphoma cell line BL-41 led to strong induction of bax-alpha, a death promoting member of the bcl-2 family, and subsequently to induction of apoptosis, suggesting a potential regulatory role of bax-alpha in sIgM-mediated cell death. In contrast, apoptosis-resistant subclones of this cell line showed only weak bax alpha expression, which was not inducible by sIgM cross-linking. In this study, we were able to demonstrate the functional significance of this observation. We stably transfected bax-alpha into a BL-41 subline resistant against sIgM-mediated apoptosis. Several bax-alpha overexpressing clones could be selected, which all showed enhanced sensitivity for sIgM-mediated apoptosis. In contrast, no sensitive clone could be identified in a large number of mock controls. This clearly indicates that induction of bax-alpha is a critical regulatory step, which sensitizes B cells for sIgM-mediated apoptosis. PMID- 9341793 TI - Down-regulation of LFA-1-mediated T cell adhesion induced by the HIV envelope glycoprotein gp160 requires phosphatidylinositol-3-kinase activity. AB - Human immunodeficiency virus binds to CD4+ T lymphocyte by the interaction, in part, between its gp120 envelope glycoprotein and the CD4 molecule. We and others have reported that the lipid kinase phosphatidylinositol-3-kinase (PI3-kinase) is associated with the CD4-p56lck complex and can be activated by various CD4 ligands. In a previous report we showed that the gp160 envelope down-regulates lymphocyte function-associated antigen-1 (LFA-1)-dependent adhesion between CD4+ T cells and B cells. This down-regulation was shown to be p56lck-dependent. Here we investigate the role of PI3-kinase in the inhibition of adhesion induced by gp160 binding to CD4. We found that gp160 activates the PI3-kinase of HUT78 CD4+ T cell lines in a way dependent on CD4-p56lck association, since no activation was detected when the interaction between CD4 and p56lck was disrupted. It was also shown, using different inhibitors of the PI3-kinase (wortmannin, Ly294002 and antisense oligonucleotides), that this lipid kinase was necessary for the down-regulation of LFA-1-mediated adhesion induced by gp160. These results strongly suggest that PI3-kinase activation induced by gp160 leads to down regulation of LFA-1-mediated T cell adhesion to B cells. Inhibition by gp160 of cytoskeleton rearrangement-dependent, anti-CD3-mediated T cell adhesion to B cells was blocked by neutralization of PI3-kinase activity, while inhibition of cytoskeleton rearrangement-independent, Mg(2+)-induced T cell adhesion was not. These results emphasize the role of PI3-kinase in the regulation of cytoskeleton structure. It is proposed that gp160 activates both p56lck and PI3-kinase which lead to a cytoskeleton organization unfavorable for LFA-1 function. PMID- 9341859 TI - Genetic studies of the leptin receptor gene in morbidly obese French Caucasian families. AB - Family studies have shown that in some populations up to 75% of the variation of body mass index can be explained by genetic factors. However, in humans, no major obesity gene has been identified to date. In contrast, there are a number of genetically well defined animal models for obesity. In two of those models (ob/ob and db/db), defects in the same pathway are responsible for obesity. Recently, some evidence has been found for the OB gene also being involved in human obesity. In this study we investigated the potential role of the OB receptor (OBR) in the etiology of massive obesity in humans using familial linkage analyses and case-control association studies. The typing of two microsatellite markers (D1S198 and D1S209), flanking the OBR gene, in 256 sib pairs showed no evidence for linkage with obesity. In order to be able to detect small gene effects, association studies with a 3'-UTR insertion/deletion polymorphism were carried out. The results of these analyses remained non-significant (chi 2 = 3.442, P = 0.18). However, subjects heterozygous for the insertion/deletion polymorphism showed a slight trend towards lower insulin values 30 min after an oral glucose load compared to homozygous individuals (P = 0.02). In summary, our results do not support a major role of the human OBR gene in the development of morbid obesity in our population. PMID- 9341860 TI - Characterisation of human patched germ line mutations in naevoid basal cell carcinoma syndrome. AB - Mutations in the human patched gene have recently been detected in patients with naevoid basal cell carcinoma syndrome. We have characterised a further 5 novel germ line mutations in patients presenting with multiple odontogenic keratocysts. Four mutations cause premature stop codons and one mutation results in an amino acid substitution towards the carboxyl terminus of the predicted patched protein. No obvious genotype-phenotype correlations could be interpreted, consistent with previous studies. PMID- 9341862 TI - Mutational analysis of ectopic factor VIII transcripts from hemophilia A patients: identification of cryptic splice site, exon skipping and novel point mutations. AB - Mutational analysis of the gene for clotting factor VIII is complicated by its large size, the high frequency of de novo mutations and its tissue-specific expression. In order to facilitate the search for mutations, we have used a combination of reverse transcription-polymerase chain reaction (RT-PCR) of ectopic factor VIII transcripts, PCR of genomic DNA, single-strand conformation polymorphism analysis and direct sequencing. Here we describe the characterization of seven potentially pathogenic mutations: five of them are novel and the reason for the pathogenicity of the sixth could be determined. Here cDNA analysis revealed the absence of the first 47 bp of exon 16 in approximately 80% of the processed factor VIII mRNA, likely due to activation of a cryptic acceptor splice site within exon 16. The other novel mutations reported here include the skipping of exon 19, which predicts the removal of the corresponding 39 amino acids from the A3 domain, and four missense mutations: W14G, Y620C, W1889L, and Q2087R. PMID- 9341861 TI - Screening for FMR1 and FMR2 mutations in 222 individuals from Spanish special schools: identification of a case of FRAXE-associated mental retardation. AB - Fragile X syndrome is the most common inherited form of familial mental retardation. It results from a (CGG)n trinucleotide expansion in the FMR1 gene leading to the typical Martin-Bell phenotype. Clinical features vary depending on age and sex. Expansion of a (CCG)n repeat in the FMR2 gene corresponds to the FRAXE fragile site which lies distal to FRAXA and is also associated with mental retardation, but it is less frequent and lacks a consistent phenotype. Analysis of repeat expansions in these two genes allows the molecular diagnosis of these different entities. We report here the screening of the FRAXA and FRAXE mutations in 222 unrelated mentally retarded individuals attending Spanish special schools. PCR and/or Southern blotting methods were used. We detected full mutations in the FMR1 gene in 11 boys (4.9%) and 1 boy (0.5%) with a CCG repeat expansion in the FMR2 gene. The latter shows mild mental retardation with psychotic behaviour and no remarkable physical traits. Molecular studies revealed a mosaicism for methylation in the FMR2 gene. This case supports the observation that expansions greater than 100 repeats can be partially methylated and cause the phenotype. PMID- 9341863 TI - Screening of the C677T mutation on the methylenetetrahydrofolate reductase gene in French patients with neural tube defects. AB - We report the analysis of the distribution of the C677T mutation on the methylenetetrahydrofolate reductase (MTHFR) gene in prenatally diagnosed neural tube defects (NTD) cases and controls. In contrast to previous reports, we found the same distribution in fetuses with NTD and controls, which suggests that the MTHFR C677T mutation cannot be regarded as a genetic risk factor for NTD. PMID- 9341864 TI - Isolation and characterization of new microsatellites at the nm23-H1 and nm23-H2 gene loci and application for loss of heterozygosity (LOH) analysis. AB - The nm23-H1 gene has been suggested to be a metastasis suppressor gene. Studies about the events of loss of heterozygosity (LOH) at the nm23 locus and its correlation to metastasis are controversially discussed. To optimize detection of LOH at the nm23 locus, we screened two P1 clones for additional microsatellites. Tumor and normal DNA from 37 colorectal, 16 gastric, and 8 germ cancer patients were examined for LOH. We found two new CA repeats, one 5' to nm23-H1 and another 3' to nm23-H2. Using these nm23 locus-specific CA repeats and five other chromosome 17 loci (D17S1522, D17S1566, D17S855, D17S515, and TP53), allele loss was observed in 4/32 (12.5%) patients with colon cancer, 2/14 (14.3%) with gastric cancer, and 1/7 (14%) with germ cancer. No isolated LOH of the nm23 region was observed. PMID- 9341865 TI - Molecular genetic analysis of two families with keratosis follicularis spinulosa decalvans: refinement of gene localization and evidence for genetic heterogeneity. AB - X-linked keratosis follicularis spinulosa decalvans (KFSD) is a rare disorder affecting both skin and eyes. In the two extended KFSD families analysed to date, the gene was mapped to Xp22.13-p22.2. By analyzing several new markers in this region, we were able to narrow the candidate region to a 1-Mb interval between DXS7161 and (DXS7593, DXS7105) in the large Dutch pedigree. In addition, we analyzed 23 markers in Xp21.2-p22.2 in a German family with KFSD. Haplotype and recombination analysis positioned the KFSD gene in this family most likely outside the candidate region on Xp22.13-p22.2. This finding is suggestive for genetic heterogeneity: in this pedigree there is either another locus on the X chromosome, or KFSD is transmitted here as an autosomal dominant trait with variable expression. PMID- 9341866 TI - Localisation and distance between ABL and BCR genes in interphase nuclei of bone marrow cells of control donors and patients with chronic myeloid leukaemia. AB - Quantitative measurements of the nuclear localisation of the ABL and BCR genes and the distance between them were performed in randomly oriented bone marrow cells of control donors and patients with chronic myeloid leukaemia (CML). Most ABL and BCR genes (75%) are located at a distance of 20-65% of the local radius from the nuclear centre to the nuclear membrane. A chimeric BCR-ABL gene located on a derivative chromosome 22 resulting from t(9;22)(q34;q11) [the so-called Philadelphia (Ph) chromosome] as well as the intact ABL and BCR genes of patients suffering from chronic myeloid leukaemia are also located mostly in this region, which has a mean thickness of 2 microns in bone marrow cells. We have not found any significant differences in the location of the two genes in the G1 and G2 phases of the cell cycle, nor between bone marrow cells and stimulated lymphocytes. Irradiation of lymphocytes with a dose of 5 Gy of gamma-rays results in a shift of both genes to the central region of the nucleus (0-20% of the radius distant from the nuclear centre) in about 15% of the cells. The minimum distance between one ABL and one BCR gene is less than 1 micron in 47.5% of bone marrow cells of control donors. Such a small distance is found between homologous ABL and between homologous BCR genes in only 8.1% and 8.4% of cells, respectively. It is possible that the relative closeness of nonhomologous ABL and BCR genes in interphase nuclei of bone marrow cells could facilitate translocation between these genes. In 16.4% of bone marrow cells one ABL and one BCR gene are juxtaposed (the distance between them varies from 0-0.5 micron) and simulate the Ph chromosome. This juxtaposition is the result of the projection of two genes located one above another into a plane, as follows from the probability calculation. PMID- 9341868 TI - A candidate gene for hemochromatosis: frequency of the C282Y and H63D mutations. AB - The gene whose alteration causes hereditary hemochromatosis (HFE according to the international nomenclature) was, more than 20 years ago, shown to map to 6p21.3. It has since escaped all efforts to identify it by positional cloning strategies. Quite recently, a gene named HLA-H was reported as being responsible for the disease. Two missense mutations, Cys282Tyr (C282Y) and His63Asp (H63D), were observed, but no proof was produced that the gene described is the hemochromatosis gene. To validate this gene as the actual site of the alteration causing hemochromatosis, we decided to look for the two mutations in 132 unrelated patients from Brittany. Our results indicate that more than 92% of these patients are homozygous for the C282Y mutation, and that all 264 chromosomes but 5 carry either mutation. These findings confirm the direct implication of HLA-H in hemochromatosis. PMID- 9341867 TI - Y chromosome polymorphisms in native American and Siberian populations: identification of native American Y chromosome haplotypes. AB - We have initiated a study of ancient male migrations from Siberia to the Americas using Y chromosome polymorphisms. The first polymorphism examined, a C-->T transition at nucleotide position 181 of the DYS199 locus, was previously reported only in Native American populations. To investigate the origin of this DYS199 polymorphism, we screened Y chromosomes from a number of Siberian, Asian, and Native American populations for this and other markers. This survey detected the T allele in all five Native American populations studied at an average frequency of 61%, and in two of nine native Siberian populations, the Siberian Eskimo (21%) and the Chukchi (17%). This finding suggested that the DYS199 T allele may have originated in Beringia and was then spread throughout the New World by the founding populations of the major subgroups of modern Native Americans. We further characterized Native American Y chromosome variation by analyzing two additional Y chromosome polymorphisms, the DYS287 Y Alu polymorphic (YAP) element insertion and a YAP-associated A-->G transition at DYS271, both commonly found in Africans. We found neither African allele associated with the DYS199 T allele in any of the Native American or native Siberian populations. However, we did find DYS287 YAP+ individuals who harbored the DYS199 C allele in one Native American population, the Mixe, and in one Asian group, the Tibetans. A correlation of these Y chromosome alleles in Native Americans with those of the DYS1 locus, as detected by the p49a/p49f (p49a,f) probes on TaqI-digested genomic DNA, revealed a complete association of DYS1 alleles (p49a,f haplotypes) 13, 18, 66, 67 and 69 with the DYS199 T allele, while DYS1 alleles 8 and 63 were associated with both the DYS199 C and T allele. PMID- 9341869 TI - Major histocompatibility complex (MHC) class III genetics in two Amerindian tribes from southern Brazil: the Kaingang and the Guarani. AB - Population genetic studies of the major histocompatibility complex (MHC) class III region, comprising C2, BF and C4 phenotypes, and molecular genetic data are rarely available for populations other than Caucasoids. We have investigated three Amerindian populations from Southern Brazil: 131 Kaingang from Ivai (KIV), 111 Kaingang (KRC) and 100 Guarani (GRC) from Rio das Cobras. Extended MHC haplotypes were derived after standard C2, BF, C4 phenotyping and restriction fragment length polymorphism (RFLP) analysis with TaqI, together with HLA data published previously by segregation analysis. C2 and BF frequencies corresponded to other Amerindian populations. C4B*Q0 frequency was high in the GRC (0.429) but low in the Kaingang. Unusual C4 alleles were found, viz. C4A*58, A*55 and C4B*22 (presumably non-Amerindian) and aberrant C4A*3 of Amerindian origin occurring with a frequency of 0.223 in the GRC. C4A*3 bands of homo- and heterozygous individuals carrying this variant were Rodgers 1 positive and Chido 1,3 positive, showed a C4A specific lysis type and a C4A like alpha-chain. Polymerase chain reaction studies and sequencing showed that this is based on a C4A*3 duplication with a regular C4A*3 and a partially converted C4A*0304 carrying the C4B specific epitopes Ch 6 and Ch 1,3. Associations of class III haplotypes with particular RFLP patterns were similar to those reported for Caucasoids. The previously described association between combined C4A and CYP21P deletions and the 6.4 kb TaqI fragment was not seen in these Amerindians. This fragment occurred within a regular two locus gene structure in the Kaingang, representing a "short" gene at C4 locus I. C4 and CYP21 duplications were frequently observed. The distribution of extended MHC haplotypes provides evidence for a close relationship between the KIV and KRC and a larger genetic distance between the two Kaingang groups and the GRC. PMID- 9341870 TI - A splicing mutation in RB1 in low penetrance retinoblastoma. AB - The pediatric eye-tumor retinoblastoma is widely held as a paradigm of human cancer genetics and has been a model system for both the two-hit hypothesis of dominantly inherited cancer as well as for the concept of tumor-specific loss of constitutional heterozygosity to achieve expression of the tumorigenic phenotype. Familial retinoblastoma is usually inherited as an autosomal dominant disease with high penetrance and expressivity. In a small but significant number of families, however, retinoblastoma is inherited with greatly reduced penetrance and expressivity. In these families, retinoblastoma tumors occur relatively late, are often unilateral, and unaffected carriers may exist. We have identified a mutation in such a family that exhibited extremely low penetrance and expressivity. This mutation appeared to affect splicing of the mutant allele such that both a normal length RB1 mRNA and a truncated RB1 mRNA were expressed from the same allele. PMID- 9341871 TI - High-throughput analysis of fragile X (CGG)n alleles in the normal and premutation range by PCR amplification and automated capillary electrophoresis. AB - Fragile X syndrome is caused by expansion of a (CGG)n trinucleotide repeat within the 5' untranslated region of the FMR1 gene transcript. The disease is reliably diagnosed by Southern blotting, but this method constitutes a significant workload and requires large samples. Therefore, for large research or screening projects in which a large majority of the samples will be normal, a more rapid and less expensive method is needed. We present a method for accurate, high throughput analysis of the FRAXA (CGG)n region in the normal and premutation range. The method is based on polymerase chain reaction (PCR) amplification of DNA extracted from whole blood or eluted from dried blood spots on filter-paper, followed by automated capillary electrophoresis and detection by multicolour fluorescence. This method allows a throughput of 144 samples in 48 h, with an intra-assay accuracy in size determination of 0.2-1.8 bp. We performed a blind reanalysis of samples from 30 patients, previously analysed by Southern blotting or PCR with radioactive labelling. In this study normal and premutation alleles, ranging from 28-121 (CGG)n repeats, were correctly determined with respect to number of (CGG)n repeats. All full-mutation alleles and one large premutation allele in a sample of a heterozygote failed to amplify. The method was used to determine the distribution of FRAXA (CGG)n repeats in the Danish population. PMID- 9341872 TI - AFX1 and p54nrb: fine mapping, genomic structure, and exclusion as candidate genes of X-linked dystonia parkinsonism. AB - We have mapped AFX1 and p54nrb to a yeast artificial chromosome (YAC) contig of Xq13.1 that harbors the X-linked dystonia parkinsonism (XDP) locus DYT3. AFX1 is flanked by loci DXS7116 and Il2R gamma, and p54nrb by loci DXS6673E and DXS7120. The exon-intron structure of both genes was analyzed. AFX1 is composed of three exons with most of exon 3 being untraslated. p54nrb is made up of 12 exons ranging in size from 40 bp to 1227 bp. The start codon is in exon 3 and the stop codon in exon 12. Both genes are expressed in the brain, among other tissues. AFX1 and p54nrb were excluded as candidates of DYT3 by sequencing of the exons and the flanking intronic sequences in an XDP patient and a control, and by Northern blot analysis. PMID- 9341874 TI - Molecular analysis of survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes of spinal muscular atrophy patients and their parents. AB - We have assayed deletions of two candidate genes for spinal muscular atrophy (SMA), the survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes, in 101 patients from 86 Chinese SMA families. Deletions of exons 7 and 8 of the telomeric SMN gene were detected in 100%, 78.6%, 96.6%, and 16.7%, in type I, II, III, and adult-onset SMA patients, respectively. Deletion of exon 7 only was found in eight type II and one type III patient. One type II patient did not have a deletion of either exon 7 or 8. The prevalence of deletions of exons 5 and 6 of the NAIP gene were 22.5% and 2.4% in type I and II SMA patients, respectively. We also examined four polymorphisms of SMN genes and found that there were only two, SMN-2 and CBCD541-2, in Chinese subjects. In our study, analysis of the ratio of the telomeric to centromeric portion (T/C ratio) of the SMN gene after enzyme digestion was performed to differentiate carriers, normals, and SMA patients. We found the T/C ratio of exon 7 of the SMN gene differed significantly among the three groups, and may be used for carrier analysis. An asymptomatic individual with homozygous deletion of exons 7 and 8 of the SMN gene showed no difference in microsatellite markers in the SMA-related 5q11.2-5q13.3. In conclusion, SMN deletion in clinically presumed child-onset SMA should be considered as confirmation of the diagnosis. However, adult-onset SMA, a heterogeneous disease with phenotypical similarities to child-onset SMA, may be caused by SMN or other gene(s). PMID- 9341873 TI - Autosomal recessive long-QT syndrome (Jervell Lange-Nielsen syndrome) is genetically heterogeneous. AB - Jervell Lange-Nielsen syndrome (JLNS) is a recessive disorder with congenital deafness and long-QT syndrome (LQTS 1). Mutations in the potassium-channel gene KVLQT1 (LQTS 1) have been identified in JLNS and in autosomal-dominant LQTS as well. We performed haplotype analysis with microsatellite markers in a Lebanese family with JLNS, but failed to detect linkage at LQTS 1. Moreover, using this approach, we excluded two other ion-channel genes involved in autosomal-dominant LQTS, HERG (LQTS 2) and SCN5A (LQTS 3). Our findings indicate that JLNS is genetically heterogeneous and that, in this family, an unknown LQTS gene causes the disease. PMID- 9341875 TI - Genetic variation of the 5-HT2A receptor gene and bipolar affective disorder. AB - Abnormalities of the serotonergic system have classically been associated with the origin of affective disorders through the biochemical action of therapeutic agents and their role in affective and perceptual states. In the present study, we hypothesized that genetic variation in the 5-hydroxytryptamine (serotonin) type 2A (5-HT2A) receptor gene (HTR2A) might have an effect on the aetiology of bipolar affective disorder. Four different polymorphisms in the HTR2A gene were studied in 88 patients with bipolar affective disorder and 113 healthy controls, all of Spanish origin. No significant association was observed between any of the four polymorphisms at the HTR2A locus, whether tested individually or as haplotypes, and bipolar affective disorder. The lack of association suggests that HTR2A is not a major risk factor for bipolar affective disorder. PMID- 9341876 TI - A novel double nucleotide substitution in the HMG box of the SRY gene associated with Swyer syndrome. AB - We describe a novel double nucleotide substitution in the SRY gene of a 46,XY female with gonadal dysgenesis or Swyer syndrome. The SRY sequence was analysed by both the single-strand conformational polymorphism assay and direct DNA sequencing of products from the polymerase chain reaction. A double nucleotide substitution was identified at codon 18 of the conserved HMG box motif, causing an arginine to asparagine amino acid substitution. The altered residue is situated in the high mobility group (HMG)-related box of the SRY protein, a potential DNA-binding domain. Since the mutation abolishes one HhaI recognition site, the results were confirmed by HhaI restriction mapping. No other mutations were found in the remaining regions of the gene. The corresponding DNA region from the patient's brother was analysed and found to be normal. We conclude that the SRY mutation in the reported XY female occurred de novo and is associated with sex reversal. PMID- 9341877 TI - The spatial distribution of human immunoglobulin genes within the nucleus: evidence for gene topography independent of cell type and transcriptional activity. AB - The three-dimensional positioning of immunoglobulin (Ig) genes within the nucleus of human cells was investigated using in situ hybridization and confocal microscopy. The visualization of heavy and light chain genes in B-lymphoid cells showed that the three Ig genes are differentially and nonrandomly distributed in different nuclear subvolumes: the kappa genes were found to be preferentially confined to an outer nuclear volume, whereas the gamma and lambda genes consistently occupied more central positions within the nucleus, the lambda genes being more interior when compared with the gamma genes. The data further show that these overall topographical distributions are independent of gene transcriptional activity and are conserved in different cell types. Although subtle gene movements within those defined topographical regions cannot be excluded by this study, the results indicate that tissue specificity of gene expression is not accompanied by drastic changes in gene nuclear topography, rather suggesting that gene organization within the nucleus may be primarily dependent on structural constraints imposed on the respective chromosomes. PMID- 9341878 TI - A third neurofibromatosis type 1 (NF1) pseudogene at chromosome 15q11.2. AB - Sequences related to the neurofibromatosis type 1 (NF1) gene have been identified on several human chromosomes. In the centromeric region of chromosomes 14 and 15, two NF1 pseudogenes have been described. Sequence comparison between NF1-related exons amplified from two yeast artificial chromosome clones hybridizing to chromosomal region 15q11.2 and published NF1-related sequences localized at 15q11.2 suggested that a third NF1 pseudogene resides in this chromosomal region. The previous localization of an NF1-related locus to the telomeric part of chromosome 15 could not be confirmed by us. Our findings further support pericentromeric spreading of partial NF1 gene copies at chromosome 15q11.2 during evolution. PMID- 9341879 TI - Novel germline mutations in the APC gene and their phenotypic spectrum in familial adenomatous polyposis kindreds. AB - Among 23 germline mutations identified in the APC screening of 45 familial adenomatous polyposis (FAP) patients, we have found 10 different novel frameshift mutations in 11 apparently unrelated patients. In two cases, an additional missense mutation was detected. One previously described as a causative germline mutation (S2621C), associated with a 1-bp insertion (4684insA) on the opposite allele, did not segregate with the FAP phenotype in the family and was therefore considered as being non-pathogenic. The other (Z1625H) was located 2 codons before a 1-bp deletion (4897delC). Both mutations were transmitted together from an FAP father to his affected son. The FAP phenotype of these 10 novel truncating mutations was clinically documented within their kindreds. Important variability was observed in the phenotype. Interestingly, we noted that a mutation (487insT) localized at the boundary of the 5' attenuated APC phenotype region in two unrelated families resulted in classical polyposis. A clear-cut genotype phenotype correlation could be drawn in only two instances. In one family, a 4684insA mutation led to a mild polyposis associated with early inherited osteomas and, in the family bearing the double mutation (Z1625H + 4897delC), the phenotype was obviously a 3' attenuated type. Our data illustrate the wide genetic and phenotypic heterogeneity of this condition between and within the families, making the establishment of correlations complex and any prediction in this disease difficult, although targeting the mutation site may be helpful in some specific cases. PMID- 9341880 TI - Two SRY-negative XX male brothers without genital ambiguity. AB - We report a Mexican family in which two sibs were identified as "classic" XX males without genital ambiguities. Molecular studies revealed that both patients were negative for several Y sequences, including SRY. A review of familial cases disclosed that this is the first family where a complete male phenotype was observed in Y-negative XX male non-twin brothers. These data suggest that an inherited loss-of-function mutation, in a gene participating in the sex determining cascade, can induce normal male sexual differentiation in the absence of SRY. PMID- 9341881 TI - The germinal center kinase gene and a novel CDC25-like gene are located in the vicinity of the PYGM gene on 11q13. AB - Multiple endocrine neoplasia type 1 (MEN1) is tightly linked to the muscle-type glycogen phosphorylase (PYGM) gene in 11q13. This region of the human genome contains additional disease-related loci implicated in the development of insulin dependent diabetes mellitus, familial paraganglioma type 2, spinocerebellar ataxia type 5, Bardet-Biedl syndrome and translocation t(11;17) described in B cell non-Hodgkin's lymphoma. We approached cloning of candidate disease genes from 11q13 by large-scale genomic sequencing. We obtained > 106 kb of sequence around the PYGM gene and established a transcriptional map that includes: (i) two genes previously localized to 11q13, PYGM and a zinc-finger protein (ZFM1) gene; (ii) the germinal center kinase (GCK, human B-lymphocyte serine/threonine protein kinase) gene; (iii) a novel human CDC25-like (HCDC25L) gene; (iv) a dystrophia myotonica protein kinase-like (DMPKL) gene; and (v) a novel ubiquitously expressed gene of unknown function (germinal center kinase- neighboring gene, GCKNG). PMID- 9341882 TI - Another pedigree with pure autosomal dominant spastic paraplegia (AD-FSP) from Tibet mapping to 14q11.2-q24.3. AB - The mutant in a family with autosomal-dominant spastic paresis in Northern Tibet was mapped by linkage analysis with several microsatellite markers to a gene locus at 14q11.2-q24.3, an area to which a few mutants leading to a condition with similar clinical signs have previously been mapped. The mutant observed in this pedigree probably arose de novo. Gene loci at 2p21-p24 and 15q, which have been found for other pedigrees with dominant spastic paresis, were excluded. The data in this pedigree do not contradict the hypothesis proposed by another group that there might be anticipation. PMID- 9341883 TI - X chromosome inactivation and micronuclei in normal and Turner individuals. AB - Studies on aneuploidy have shown that the X is the most frequently lost chromosome in females, and that the number of X chromosome-positive micronuclei increases with age in women. Recently, we showed that the inactive X chromosome is incorporated preferentially in micronuclei. The objectives of the current study were, firstly, to determine the incidence of X chromosome incorporation into micronuclei in males and, secondly, to determine the incidence of X chromosome incorporation into micronuclei of females with Turner syndrome. Blood samples were obtained from 18 male newborns and 35 normal adult males ranging in age from 22 to 79 years and from seven women with non-mosaic Turner syndrome aged 11-39 years. Isolated lymphocytes were cultured in the presence of cytochalasin B and 2000 binucleated cells per subject were scored for micronuclei. Cells were then hybridized with the biotinylated X centromere-specific probe, pBamX7, and visualized with fluorescein-conjugated avidin. All micronucleated cells were relocated and evaluated for the presence or absence of the X chromosome. Of the 335 micronuclei observed, 6.6% (22/335) contained an X chromosome. Analysis of variance shows a statistically significant increase, for both males and Turner females, in the number of X chromosome-positive micronuclei with age (P < 0.001). These data also show that the X chromosome is included in micronuclei from males more often than would be expected by chance (P < 0.005; chi 2 analysis, 15 df). Here we show that there is a tenfold difference in the frequency of X chromosome positive micronuclei in 46,XX females compared to 46,XY males and 45,X females, providing further support to our previous finding that the X chromosome in micronuclei is the inactive chromosome. PMID- 9341884 TI - Frequent gains on chromosome arms 1q and/or 8q in human endometrial cancer. AB - Comparative genomic hybridization (CGH) was employed to survey genomic regions with increased and decreased copy number of the DNA sequence in 15 endometrial cancers [10 cases with microsatellite instability positive (MI+) and 5 cases with MI-]. Twelve of these 15 tumors (80%) showed abnormalities in copy number at one or more of the chromosomal regions. There were no regions with frequent chromosomal losses. Conversely, 11 of 15 cases (73%) showed gains on chromosome arms 1q (8/15; 53%) and/or 8q (6/15; 40%). Concordant gains of both chromosome arms 1q and 8q were observed in all three endometrial cancers of histological grade 3. These results suggest that these two chromosomal regions may contain genes whose increased expression contributes to development and/or progression of endometrial carcinogenesis. Two cases were further analyzed by fluorescence in situ hybridization (FISH) using three probes on chromosome 1 and two probes on chromosome 8 to more accurately determine increases in copy number. We found gains of chromosome 1q to 2.9-3.6 copies per cell and on 8q to 4.4 copies per cell. PMID- 9341885 TI - Molecular analysis of dihydropteridine reductase deficiency: identification of two novel mutations in Japanese patients. AB - Mutations in the dihydropteridine reductase (DHPR) gene result in hyperphenylalaninaemia and deficiency of various neurotransmitters in the central nervous system, causing severe neurological symptoms. We studied two Japanese patients with DHPR deficiency and identified a missense and a splicing error mutation, respectively. A homozygous missense mutation (tryptophan36-to-arginine) was detected in patient 1. The mutation abolished DHPR activity according to in vitro expression studies. The DHPR mRNA in patient 2 was markedly decreased. Reverse transcription-polymerase chain reaction of the mRNA generated a cDNA fragment with a 152-bp insertion. The inserted sequence contained a termination codon, which was likely to affect the stability of the mRNA. Analysis of genomic DNA showed that the insertion was derived from putative intron 3 of the DHPR gene, and an intronic A-to-G substitution was present adjacent to the 3'-end of the inserted sequence. The nucleotide change generated a sequence similar to an RNA splice donor site and probably activated an upstream cryptic acceptor site, thus producing an abnormal extra exon. PMID- 9341886 TI - Detection and characterization of mitochondrial DNA rearrangements in Pearson and Kearns-Sayre syndromes by long PCR. AB - We used a strategy based on long PCR (polymerase chain reaction) for detection and characterization of mitochondrial DNA (mtDNA) rearrangements in two patients with clinical signs suggesting Pearson syndrome and Kearns-Sayre syndrome (KSS), respectively, and one patient with myopathic symptoms of unidentified origin. Mitochondrial DNA rearrangements were detected by amplification of the complete mitochondrial genome (16.6 kb) using long PCR with primers located in essential regions of the mitochondrial genome and quantified by three-primer PCR. Long PCR with deletion-specific primers was used for identification and quantitative estimation of the different forms of rearranged molecules, such as deletions and duplications. We detected significant amounts of a common 7.4-kb deletion flanked by a 12-bp direct repeat in all tissues tested from the patient with Pearson syndrome. In skeletal muscle from the patient with clinical signs of KSS we found significant amounts of a novel 3.7-kb rearrangement flanked by a 4-bp inverted repeat that was present in the form of deletions as well as duplications. In the patient suffering from myopathic symptoms of unidentified origin we did not detect rearranged mtDNA in blood but found low levels of two rearranged mtDNA populations in skeletal muscle, a previously described 7-kb deletion flanked by a 7-bp direct repeat and a novel 6.6-kb deletion with no repeat. These two populations, however, were unlikely to be the cause of the myopathic symptoms as they were present at low levels (10-40 ppm). Using a strategy based on screening with long PCR we were able to detect and characterize high as well as low levels of mtDNA rearrangements in three patients. PMID- 9341887 TI - Type 2 oculocutaneous albinism (OCA2) in Zimbabwe and Cameroon: distribution of the 2.7-kb deletion allele of the P gene. AB - In previous studies, we characterized a 2.7-kb interstitial deletion allele of the P gene associated with tyrosinase-positive oculocutaneous albinism (OCA2) in African Americans and Africans. In this study, we investigated the frequency of this allele among OCA2 subjects in two African countries, Zimbabwe and Cameroon. The deletion allele was most common in Zimbabwe, comprising nearly all (92%) mutant alleles, which is the highest incidence reported so far. In addition, the deletion allele was widespread but less common among OCA2 Cameroonians and accounted for 65% of the mutant alleles. PMID- 9341888 TI - The European Consortium on MEN1. Linkage disequilibrium studies in multiple endocrine neoplasia type 1 (MEN1). PMID- 9341889 TI - Expected effects of mass screening policies on the frequency of cystic fibrosis homozygotes. AB - We evaluated, by deterministic computer simulation, some effects of a screening programme for carriers of cystic fibrosis mutations. Two different selective regimes (heterozygote advantage and directional selection against recessive homozygotes) and three kinds of response to the screening were simulated. The curves describing the expected decline in the frequency of CF homozygotes allow one to predict some benefits of a screening campaign. In addition, it is shown that a strategy aimed at testing couples, rather than individuals, may become less expensive after only two generations of screening. The main source of uncertainty for a screening programme remains the selection mechanism, namely the existence of some sort of biological advantage for heterozygous carriers of CF mutations. PMID- 9341890 TI - A large family with subtelomeric translocation t(18;21)(q23;q22.1) and molecular breakpoint in the Down syndrome critical region. AB - We describe a 17-month-old infant with clinical features of Down syndrome and a normal karyotype by standard chromosomal analysis, her two uncles aged 28 and 30 years, respectively, with reduced intelligence and unusual appearance but not apparent Down syndrome, and a severely retarded 6-year-old girl with dysmorphy and epilepsy from the same family. Cytogenetic studies of patients and normal intervening relatives had been carried out at different institutions with normal results. Fluorescence in situ hybridization using whole chromosome painting and unique-copy probes (cosmids) and high-resolution banding revealed a familial subtelomeric translocation of chromosomes 18 and 21, resulting in partial trisomy 21 in the infant and her two uncles, and partial monosomy 21 in the 6-year-old girl. Cytogenetic breakpoints were located in bands 18q23 and 21q22.1, respectively. The molecular breakpoint on chromosome 21 was located between D21S211 (proximal) and D21S1283 (distal) and thus maps within the Down syndrome critical region. PMID- 9341891 TI - Different entities of proximal spinal muscular atrophy within one family. AB - The molecular analysis of the survival motor neuron (SMN) gene and several closely flanking polymorphic markers in an atypical pedigree with four patients suffering from spinal muscular atrophy (SMA) over two generations has raised new aspects concerning the etiology and the molecular spectrum of autosomal recessive SMA. Three patients in two generations show homozygous deletions of exons 7 and 8 of the telomeric copy of SMN (telSMN), thus confirming the presence of autosomal recessive SMA, with localisation on chromosome 5q12. The fourth SMA patient with mild neurogenic atrophy (confirmed by muscle biopsy and electromyography) shows no homozygous deletion of telSMN but carries a heterozygous deletion of telSMN, as can be deduced from her two affected homozygously deleted children. No intragenic mutation has been identified in the remaining telSMN. In addition, she shares only one SMA chromosome with her affected brother, is haploidentical with two healthy brothers, and has a 31-year-old healthy son, who has inherited an SMN deleted paternal chromosome and the SMN non-deleted maternal chromosome. These results suggest that this patient either has a neurogenic atrophy of a different origin or exhibits an unusual heterozygous manifestation of SMA 5q12. Interestingly, the two haploidentical telSMN-deleted affected sibs in the second generation show a strikingly discordant clinical picture indicating that, in addition to telSMN mutations, other factors influence the phenotype of SMA in the reported pedigree. PMID- 9341892 TI - New p57KIP2 mutations in Beckwith-Wiedemann syndrome. AB - Beckwith-Wiedemann syndrome (BWS) is characterized by numerous growth abnormalities and an increased risk of childhood tumors. The gene for BWS is localized in the 11p15.5 region, as determined by linkage analysis of autosomal dominant pedigrees. The increased maternal transmission pattern seen in the autosomal dominant-type pedigrees and the findings of paternal uniparental disomy reported for a subgroup of patients indicate that the gene for BWS is imprinted. Previously, we found p57KIP2, which is a Cdk-kinase inhibitor located at 11p15, is mutated in two BWS patients. Here, we screened for the mutation of the gene in 15 BWS patients. PMID- 9341893 TI - Isolation by distance in Germany. PMID- 9341894 TI - "Salvage chemotherapy" of malignant disease: importance of precisely defining the goals of treatment. PMID- 9341895 TI - Tumour regression in non-small-cell lung cancer following neoadjuvant therapy. Histological assessment. AB - In the scope of a prospective multi-centre study after neoadjuvant combined chemotherapy (carboplatin, ifosfamide, etoposide, vindesine) and radiotherapy (45 Gy) 40 resection specimens of locally advanced non-small-cell lung cancer were analysed in order to establish reproducible pathological/anatomical results of tumour regression. Resection specimens of 28 squamous cell carcinomas and 12 adenocarcinomas were investigated using serial sections of the primary lesion. The mean age of the patients was 57 years. The results were compared to spontaneous regressive changes in a control group of 50 untreated non-small-cell lung cancers. Marked scarry fibrosis in the region of the former primary tumour, concentric foci of fresh tumour necroses and surrounding foam cell clusters with transition into vascular granulation tissue could be established as characteristic features of therapy-induced tumour regression, whereas untreated carcinomas revealed necroses with adjoining vital tumour tissue. Using a three step regression system, 3 tumours could be classified as grade I (no or only slight tumour regression), 10 tumours as grade IIA (marked but incomplete tumour regression, more than 10% vital tumour tissue), 20 tumours as grade IIB (less than 10% vital tumour tissue) and 7 tumours as grade III (complete tumour regression without vital tumour tissue). After a median follow-up period of 32.3 months in patients with grade IIB or III tumour regression ("responders") the median survival time of 27.9 months was found to be significantly longer than in patients with grade I or IIA tumour regression ("non-responders") with a median survival period of 13.7 months (log-rank test, P = 0.020). The resection specimens analysed, which were obtained 7 weeks (on average) after the end of radiochemotherapy, did not show specific changes due to preoperative therapy, but quite characteristic histological alterations in the former tumour area were registered, which had been induced by combined neoadjuvant radiation and chemotherapy. The grade of therapy-induced tumour regression could be shown to be a significant prognostic factor in non-small-cell lung cancer. PMID- 9341896 TI - Augmented human-tumor-cytolytic activity of peripheral blood lymphocytes and cells from a mixed lymphocyte/tumor culture activated by interleukin-12 plus interleukin-2, and the phenotypic characterization of the cells in patients with advanced carcinoma. AB - In this study, we evaluated the ability of combination regimens of interleukin-12 (IL-12) and interleukin-2 (IL-2) to induce effective killer cells against human tumors in vitro, in peripheral blood lymphocytes (PBL) from 15 cancer patients and mixed lymphocyte/tumor culture (MLTC) cells from 16 cancer patients, and carried out a phenotypic analysis of the cells responsible for the lysis of the human tumors. The freshly prepared PBL were cultivated with IL-2 alone or IL 12/IL-2 for 10 days [lymphokine-activated killer (LAK) cell generation system]. The MLTC cells (PBL cultured with mitomycin-C-treated allogeneic G-415 tumor cells for 3 days) were further cultivated with IL-2 or IL-12/IL-2 for 7 days [cytotoxic T lymphocytes (CTL) generation system]. The cytolytic activities of the lymphoid cells cultivated with IL-12/IL-2 were significantly augmented in both the LAK and CTL generation systems, as compared with those of cells treated with IL-2 alone. In the LAK generation system, the cytolytic activities of the cells cultivated with IL-12/IL-2 were significantly decreased by the method of negative selection of CD11b- or CD56+ cells using immunomagnetic beads. The CD8(+)-depleted cells showed a slight decrease of activity. The killer cell activities of the CD4(+)-depleted cells remained unchanged. In the CTL generation system, the activity was markedly reduced by the elimination of the CD8+ or CD11b+ or CD56+ cells. The combined data suggested that IL-12/IL-2-induced killer effector cells in the LAK generation system were mainly of the natural killer (NK) type, comprising CD8-CD11b+, CD8- CD16b+, CD3-CD56+, and partly possible CD8+ CD11b- T cells. CD8+ CD11b- T cells mixed with cells of the NK type, comprising CD8-CD11b+, CD8- CD16b+ and CD3-CD56+ cells, were the population of killer effector cells induced by IL-12/IL-2 in the CTL generation system. PMID- 9341897 TI - Loss of heterozygosity of the nm23-H1 gene in human renal cell carcinomas. AB - This study evaluates the potential contribution of the nm23-H1 gene to malignant transformation in patients with renal cell carcinoma. Using specific oligonucleotide primers for the nm23-H1 microsatellite repetitive sequence, gene instability was followed by polymerase chain reaction/loss of heterozygosity assay on 54 tumor specimens and the corresponding normal tissue samples. We also determined, immunohistochemically, the relative concentration and localization of the nm23-H1 protein product. From 77.7% informative cases, DNA from 6 tumors exhibited loss of heterozygosity, regardless of the tumor stage (TNM). Out of 39 samples analyzed, 30 were negative for Nm23-H1 protein, while the others were only slightly positive. No correlation with tumor stage was found. Normal renal tissue was also negative for this protein. Our results provide the evidence for loss of heterozygosity, followed by means of microsatellite tandem-repeat polymorphism, at the nm23-H1 locus in renal cell carcinoma. However, since no correlation was found between the tumor stage or metastatic potential on the one hand, and allelic loss and specific protein expression on the other, it seems that nm23-H1 does not play a key role in the invasiveness of this tumor type. PMID- 9341899 TI - p53 is a persistent and predictive marker in advanced ovarian carcinomas: multivariate analysis including comparison with Ki67 immunoreactivity. AB - p53 mutation and p53 protein overexpression are common findings in ovarian carcinomas. In order to evaluate the prognostic significance of the p53 status and its role in metastasis, we examined 104 ovarian carcinomas, among them 83 cases with follow-up data, and 40 pairs of primary tumors and metastases, by p53 immunohistochemistry and temperature-gradient gel electrophoresis. Comparison of primary tumors and their metastases revealed identical results in 88%-90% of the cases, indicating that, in most cases, mutant p53 occurs prior to metastatic spread and remains clonally conserved. With respect to all tumors, moderate/high p53 expression was significantly more prevalent in serous-papillary types, carcinomas with high grade, and high Ki67 scores, but was not associated with age, stage, or hormone receptor status. Kaplan-Meier analysis of 83 cases, followed-up for 9-96 months, demonstrated that moderate/high p53 overexpression in the group of 66 stage T3/M1 tumors was associated significantly (P = 0.0028 and P = 0.0105) with shorter overall and recurrence-free survival. Multivariate analysis revealed that advanced clinical stage and p53 positivity were the only independent predictive variables. No significance was seen in regard to second look results and outcome of 50 patients receiving platinum-based chemotherapy. These observations show that p52 immunohistochemistry is an independent prognostic indicator at the given cut-off level, but does not reliably predict chemotherapy response. PMID- 9341898 TI - Loss of heterozygosity of the retinoblastoma gene in liver cirrhosis accompanying hepatocellular carcinoma. AB - Carcinogenesis is a multistep process. Most hepatocellular carcinoma (HCC) is preceded by liver cirrhosis, but the genetic changes involved in cirrhosis are not known well. The present study was conducted to evaluate aberration of the retinoblastoma (RB) gene in HCC and adjacent non-tumorous liver using 22 patients with chronic liver damage accompanying HCC. The specimens obtained by microdissection from paraffin-embedded tissues were analyzed using an assay based on the polymerase chain reaction for highly polymorphic nucleotide sequences of microsatellites in the RB gene. Out of 22 cases, 15 showed constitutional heterozygosity for the microsatellite markers. In 11 (73.3%) of these 15 informative cases, the primary HCC foci showed loss of heterozygosity (LOH). In 8 of these 11 doubly informative (informative and LOH-positive in primary HCC) cases, LOH was found in 20 (64.5%) of 31 microdissected non-tumorous foci. All of the non-tumorous foci showing RB loss were cirrhotic lesions but there were no foci of chronic hepatitis. The remaining 4 cases without LOH in HCC foci showed no LOH in non-tumorous lesions. In our study, LOH of the RB gene was frequently observed in liver cirrhosis surrounding tumor. PMID- 9341900 TI - Prognostic value of immunohistochemistry for p53 in primary soft-tissue sarcomas: a multivariate analysis of five antibodies. AB - Most changes of tumor suppressor p53 and its pathway involve a protein with prolonged half-life that permits immunohistochemical detection. The goal of this study was to compare the prognostic relevance of five different p53 antibodies in primary soft-tissue sarcomas (STS) with known p53 mutation status, using a multivariate Cox regression model (adjusted to tumor grading, staging, localization, tumor type, and therapy). A group of 198 primary STS of six types were investigated for p53 overexpression, using p53 antibodies DO-1, DO-7, Pab1801, Pab240, and CM-1. A positive marker frequency between 36.2% and 62.6% was detected. Out of 65 patients whose primary tumor reacted positively to all five antibodies, 52 (80%) died within the study period. Only the N-terminal binding monoclonal antibodies DO-1, DO-7 and Pab1801 showed a multivariate correlation with survival (P = 0.0014, 0.0048 and 0.02). CM-1 and Pab240 had a univariate, but not a multivariate correlation, with a confounding effect of grading. The prognostic relevance for the five p53 antibodies was: DO-1 > Pab1801 > DO-7 > CM-1 > Pab240. This is the first study that investigates multivariately the prognostic relevance of p53 immunostaining in STS. If monoclonal antibodies with an epitope in the N-terminal region of the p53 protein (DO-1, Pab1801, DO-7) are applied, p53 immunohistochemistry provides an independent prognostic marker in STS. PMID- 9341901 TI - Epstein-Barr virus in malignancies in renal transplant recipients in Japan. AB - A role for Epstein-Barr virus (EBV) in the development of malignancies including lymphomas, and carcinoma of the stomach, nasopharynx, thymus and salivary gland is suggested. It is indicated that EBV evokes polyclonal-B-cell-proliferative diseases in immunocompromised hosts, such as transplant patients, which results in monoclonal malignant lymphomas. The suppression of immune functions in these patients is thought to lead to incomplete elimination of the cells expressing EBV latent infection genes. To examine the etiological role of EBV in the development of malignancies following renal transplant in Japan, 42 malignancies in 1744 cases of renal transplant were studied for the presence and type of EBV. The polymerase chain reaction revealed that 5 malignancies were positive for EBV, all type A: 2 of 2 cases of non-Hodgkin's lymphoma (NHL), 2 of 8 cases of gastric adenocarcinoma of the common type, and 1 of 2 cases of gastric plasmacytoma. In situ hybridization revealed positive signals in the nucleus of tumor cells in 2 cases of NHL and 1 of plasmacytoma. Positive signals were found in the small lymphoid cells but not in the tumor cells in 2 cases of gastric carcinoma. On the basis of these findings, a role for EBV in the development of malignancies in renal transplant patients is unlikely except for lymphoid neoplasias. PMID- 9341903 TI - Signal transduction and mechanisms of cell death. Sixth colloquium on cellular signal transduction, 17 January 1997, Heidelberg, Germany. PMID- 9341902 TI - Reverse transcriptase/polymerase chain reaction analysis of parathyroid hormone related protein for the detection of tumor cell dissemination in the peripheral blood and bone marrow of patients with breast cancer. AB - Tumor cell dissemination in the bone marrow is an independent prognostic marker for relapse and survival for patients with primary breast cancer. Parathyroid hormone-related protein (PTHrP) is expressed in most primary tumors and bone metastases of patients with breast cancer. PTHrP acts as an autocrine growth factor for breast cancer cells in vitro and there is evidence that it is especially important for osseous metastasis. For a sensitive detection of PTHrP positive disseminated tumor cells a reverse transcriptase/polymerase chain reaction (RT/PCR) assay for PTHrP transcripts in the peripheral blood (PB) and in the bone marrow (BM) has been established. In mixing studies, the sensitivity of the reverse transcriptase/polymerase chain reaction (RT/PCR) for PTHrP was one tumor cell in 1 x 10(6) mononuclear cells. At this level of sensitivity, transcripts of PTHrP were detected in none of 30 PB samples and in 3 of 25 BM samples of healthy volunteers; there were also no transcripts of PTHrP in the PB and BM of 6 patients with benign breast lesions. The PB samples of 31 patients and the BM samples of 34 patients with predominantly early-stage breast cancer were tested for PTHrP expression along with immunocytology against cytokeratin 18 (CK18) as a standard immunological detection technique. PTHrP expression was shown in 9 of 31 patients in the PB and in 9 of 34 patients in the BM. In 30 patients, PB and BM samples were available simultaneously. There were cases of combined positive findings in the PB and the BM (4/30) and of isolated positivity in the PB (5/30) or in the BM (4/30). Compared to immunocytology, RT/PCR assay of PTHrP assay was significantly more sensitive in the peripheral blood (8/30 by RT/PCR compared to 1/30 by immunocytology). In the bone marrow there were cases of positivity for both markers (2/34), cases of isolated positivity by immunocytology for CK18 (3/34) and cases of isolated positivity for PTHrP transcripts (7/34). In conclusion the RT/PCR assay for PTHrP transcripts is a feasible and very sensitive technique for the detection of tumor cell dissemination in the PB, even in patients with early-stage breast cancer. The specificity of detection of PTHrP transcripts in the bone marrow is limited, possibly because of autochthonous expression of PTHrP in osteoblastic cells. The clinical follow-up of the subgroups of patients at risk, as defined by this assay, will show its prognostic significance for patients with breast cancer. PMID- 9341904 TI - Isozymes of nuclear protein tyrosine kinase during chemical induced hepatocarcinogenesis in rats. AB - The activity changes of isozymes of nuclear protein tyrosine kinase (nPTK) during chemically induced hepatocarcinogenesis in rats was studied. Two isozymes were isolated from rat liver by using ion exchange chromatography. These two isozymes were designated as nPTK-I and nPTK-II, and were found to be minor and major components of total nPTK respectively. These proved to be two different enzymes due to their different characteristics, such as molecular weights, electrical charges and kinetics. The specific activity nPTK-I and its percentages in total nPTK significantly increased in preneoplastic stage (week 10) and slightly elevated further in neoplastic stage (week 18). The specific activity of the nPTK II moderately increased at week 10, but compared to its value at week 10, it decrease at week 18, resulted in gradual decrease of its percentage in total nPTK. These results indicate that rat hepatic nPTK-I and nTPK-II may be related to neoplastic and preneoplastic stages of rat liver respectively. PMID- 9341905 TI - C-terminal modifications of histidyl-N-benzylglycinamides to give improved inhibition of Ras farnesyltransferase, cellular activity, and anticancer activity in mice. PMID- 9341906 TI - Probing the mechanism of action and decomposition of amino acid phosphomonoester amidates of antiviral nucleoside prodrugs. AB - The decomposition pathways in peripheral blood mononuclear cells (PBMCs) and the in vitro anti-HIV-1 activity of the structurally similar 3'-azido-3' deoxythymidine (AZT) phosphoramidates 1-6 and 3'-fluoro-3'-deoxythymidine (FLT) phosphoramidates 7-10 are reported. The AZT phosphoramidates exhibited no cytotoxicity toward CEM cells at concentrations as high as 100 microM, whereas the FLT phosphoramidates 9 and 10 had CC50 values of 95.6 and 35.1 microM, respectively. All 10 compounds exhibited no cytotoxicity toward PBMCs at concentrations as high as 100 microM and were effective at inhibiting viral replication. In particular, the AZT phosphomonoester amidate 4 displayed comparable antiviral activity to the parent nucleoside analog AZT. Mechanistic studies on the amino acid carbomethoxy ester phosphomonoester amidates revealed that their decomposition pathway differs from that of amino acid carbomethoxy ester aryl phosphodiester amidates of nucleotide prodrugs. AZT phosphomonoester amidates are internalized by lymphocytes to the same extent as AZT by a nonsaturable process. In lymphocytes, the amino acid carbomethoxy ester phosphomonoester amidates of AZT are not significantly metabolized to either AZT or the mono-, di-, or triphosphate of AZT. The amount of active anabolite, AZT-5' triphosphate, formed in PBMCs incubated with the AZT phosphomonoester amidates 3 and 4 was 2- and 3-fold less than that observed after treatment with AZT, respectively. In contrast, FLT phosphomonoester amidates are rapidly converted to FLT-5'-monophosphate by a process that is antagonized by the corresponding AZT derivative 4. These results suggest that the metabolism of aromatic amino acid carbomethoxy ester phosphomonoester amidate nucleotide prodrugs by PBMCs does not require prior conversion to the corresponding carboxylic acid before proceeding to P-N bond cleavage. PMID- 9341907 TI - Synthesis of a novel sialic acid derivative (sialylphospholipid) as an antirotaviral agent. AB - A novel sialylphospholipid (SPL) was synthesized from N-acetylneuraminic acid (NeuAc) and phosphatidylcholine (PC) by a chemical and enzymatic method and evaluated as an inhibitor of rotavirus. PC and 1,8-octanediol were conjugated by transesterification reaction of Streptomyces phospholipase D (PLD) under a water chloroform biphasic system to afford phosphatidyloctanol, which was condensed with a protected 2-chloro-2-deoxyneuraminic acid derivative by using silver trifluoromethanesulfonate as an activator in chloroform and converted, after deprotection, to SPL. Rhesus monkey kidney cells (MA-104) were incubated with simian (SA-11 strain) and human (MO strain) rotaviruses in the presence of SPL, and the cells infected were detected indirectly with anti-rotavirus antibody. SPL showed dose dependent inhibition against both virus strains. The concentrations required for 50% inhibition (IC50) against SA-11 and MO were 4.35 and 16.1 microM, respectively, corresponding to 10(3)- and 10(4)-fold increases in inhibition as compared to monomeric NeuAc. PMID- 9341909 TI - Search for DNA repair inhibitors: selective binding of nucleic bases-acridine conjugates to a DNA duplex containing an abasic site. AB - The abasic site is one of the most frequent DNA lesions generated by spontaneous or enzymatic cleavage of the N-glycosidic bond. The abasic site is also an intermediate in the nucleotide and base excision DNA repair. We examined molecules which recognize and cleave DNA at the abasic site with high efficiency. These molecules incorporate in their structure a nucleic base for abasic site recognition, an intercalator for DNA binding, and a polyamino linker for ionic interaction and DNA cleavage. Such compounds, by interfering with abasic sites in DNA, are also inhibitors of DNA repair. In order to better understand the parameters of the interaction, we carried out a UV thermal denaturation study of synthetic oligonucleotides containing the lesion both in the absence and in the presence of the drugs. A similar study was also carried out using the corresponding nonmodified oligonucleotide. The results indicate selective binding of the base-chain-intercalator conjugates to the abasic site containing oligonucleotides. PMID- 9341908 TI - Effect of a chemical modification on the hydrated adenosine intermediate produced by adenosine deaminase and a model reaction for a potential mechanism of action of 5-aminoimidazole ribonucleotide carboxylase. AB - Using the hydrated adenosine intermediate (6R)-6-amino-1, 6-dihydro-6-hydroxy-9 (beta-D-ribofuranosyl)purine (2) produced by adenosine deaminase (ADA, EC 3.5.4.4) as a starting point, the active site probe and inhibitor platform 5 (formylamino)imidazole riboside (FAIRs, 4) was designed by removal of the C6(OH)(NH2)-molecular fragment of 2 generated by the early events of the enzyme catalyzed hydrolysis. FAIRs was synthesized directly from the sodium salt of 5 amino-1-(beta-D-ribofuranosyl)imidazole-4-carboxylic acid (CAIR) along a reaction sequence involving a tandem N-formylation/decarboxylation that may have a mechanistic connection to the Escherichia coli purE-catalyzed constitutional isomerization of N5-CAIR to CAIR. The physical and spectral properties of FAIRs were elucidated, its X-ray crystal and NMR solution structures were determined, and its interaction with ADA was investigated. Crystalline FAIRs exists solely as the Z-formamide rotamer and exhibits many of the same intramolecular hydrogen bonding events known to contribute to the association of Ado to ADA. In water and various organic solvents, however, FAIRs exists as NMR-distinct, slowly interconverting Z and E rotamers. This truncated enzymatic tetrahedral intermediate analog was determined to be a competitive inhibitor of ADA with an apparent Ki binding constant of 40 microM, a value quite close to that (33 microM) of the natural substrate's K(m). The actual species selected for binding by ADA, though, is likely the minor hydroxyimino prototropic form of Z-FAIRs possessing a far lower true Ki value. As the structural features of FAIRs appear well-suited to support its use as a template for constructing active site probes of both ADA and AIR carboxylases, a variety of carbohydrate-protected versions of FAIRs suitable for facile aglycon elaborations were synthesized. The N3 alkylation, N3-borane complexation, and C4-iodination of some of these were investigated in order to assess physicochemical properties that may assist in the elucidation of mechanisms for the AIR carboxylases. The survey of these properties taken together with a reasonable mechanism for the model CAIRs-->FAIRs synthetic transformation is interpreted to support a mechanism for the purE catalyzed N5-CAIR-->CAIR biosynthetic one that involves a carboxylative sp3 rehybridization of the imidazole C4 atom rather than one possessing a dipole stabilized C4 sp2 carbanionic intermediate. PMID- 9341910 TI - Synthesis of gonadotropin-releasing hormone III analogs. Structure-antitumor activity relationships. AB - Following the observation that the activity of gonadotropin-releasing hormone III (GnRH-III) in the suppression of growth of MDA-MB-231 and MCF-7 breast cancer cells surpasses that of GnRH and other analogs thereof, analogs of GnRH-III were synthesized to investigate the structural basis for the improved antitumor activity. Compounds synthesized include analogs with changes in the central sequence in which GnRH-III differs from GnRH and in the C- and N-terminal regions. The results indicate that a salt bridge between Asp6 and Lys8 stabilizes the active conformation of GnRH-III and show the importance of the Trp7. Replacement of the C-terminal Gly-NH2 with D-Ala-NH2 was not well tolerated, but replacement with ethylamide was. Replacement of pGlu1 with Ac-D-Trp appears to have a significantly deleterious effect on a unique conformation of GnRH-III which is responsible for its binding to the receptors on cancer cell lines and the resultant antitumor activity. PMID- 9341911 TI - Potent alpha 4 beta 1 peptide antagonists as potential anti-inflammatory agents. AB - The migration, adhesion, and subsequent extravasation of leukocytes into inflamed tissues contribute to the pathogenesis of a variety of inflammatory diseases including asthma, rheumatoid arthritis, inflammatory bowel disease, and multiple sclerosis. The integrin adhesion receptor alpha 4 beta 1 expressed on leukocytes binds to the extracellular matrix protein fibronectin and to the cytokine inducible vascular cell adhesion molecule-1 (VCAM-1) at inflamed sites. Binding of alpha 4 beta 1 to VCAM-1 initiates firm adhesion of the leukocyte to the vascular endothelium followed by extravasation into the tissue. Monoclonal antibodies generated against either alpha 4 beta 1 or VCAM-1 can moderate this inflammatory response in a variety of animal models. Recently peptides containing a consensus LDV sequence based on the connecting segment-1 (CS-1) of fibronectin and cyclic peptides containing an RCD motif have shown promise in modulating leukocyte migration and inflammation presumably by blocking the interaction of alpha 4 beta 1 with VCAM-1. Here we describe novel, highly potent, cyclic peptides that competitively inhibit alpha 4 beta 1 binding to VCAM-1 and fibronectin at sub nanomolar concentrations. The structure of a representative analog was determined via NMR spectroscopy and used to facilitate optimization of peptide leads. The peptides discussed here utilize similar functional groups as the binding epitope of VCAM-1, inhibit lymphocyte migration in vivo, and are highly selective for alpha 4 beta 1. Furthermore the structure--activity relationships described here have provided a template for the structure-based design of small molecule antagonists of alpha 4 beta 1-mediated cell adhesion processes. PMID- 9341912 TI - Phenylimidazolidin-2-one derivatives as selective 5-HT3 receptor antagonists and refinement of the pharmacophore model for 5-HT3 receptor binding. AB - A possible bioisosterism between the benzamido and the phenylimidazolidin-2-one moieties has been suggested on the basis of the similarity between the molecular electrostatic potential (MEP) of metoclopramide, a D2 receptor antagonist with weak 5-HT3 receptor antagonist properties, and zetidoline, a D2 receptor antagonist. Starting from this premise, a series of phenylimidazolidin-2-one derivatives bearing a basic azabicycloalkyl or an imidazolylalkyl moiety were synthesized and evaluated for 5-HT3 receptor radioligand binding affinity ([3H] GR 43,694). In vitro 5-HT3 receptor antagonist activity was tested in the guinea pig ileum assay (GPI). A number of high-affinity ligands were shown to be potent 5-HT3 receptor antagonists in vivo as determined by inhibition of the Bezold- Jarisch reflex in the anesthetized rat. In general, the imidazolylalkyl derivatives were found to be more active than azabicycloalkyls. 1-(3,5 Dichlorophenyl)-3-[(5-methyl-1H-imidazol-4-yl)methyl]imidazoli din-2-one (58), in particular, displayed very high affinity for the 5-HT3 receptor (Ki of 0.038 nM) with a Kb of 5.62 nM in the GPI assay, being more potent than the reference compounds (ondansetron, tropisetron, granisetron, and BRL 46,470) tested. 58 showed an ID50 comparable to that of ondansetron (2.2 micrograms/kg i.v.) in the Bezold--Jarisch reflex. A molecular modeling study based on this structurally novel series of compounds allowed the refinement of previously reported 5-HT3 receptor antagonist pharmacophore models. PMID- 9341913 TI - Structure-activity relationships of (4-acylpyrrol-2-yl)alkanoic acids as inhibitors of the cytosolic phospholipase A2: variation of the substituents in positions 1, 3, and 5. AB - Derivatives of 3-(1,3,5-trimethyl-4-octadecanoylpyrrol-2-yl)propionic acid (1) and (1,3,5-trimethyl-4-octadecanoylpyrrol-2-yl)acetic acid (4) were prepared and evaluated for their ability to inhibit the cytosolic phospholipase A2 of intact bovine platelets. While replacement of one of the methyl groups in position 1, 3, or 5 of the acetic acid 4 by a benzyl residue did not influence the inhibitory potency significantly, the introduction of a dodecyl chain led to compounds which even enhanced the enzymatic activity. Stepwise elongation of the alkyl substituent in position 1 showed that the ability to inhibit the enzyme was lost when the alkyl chain exceeded a length of five carbons in case of compound 1 or six carbons in case of compound 4. Introduction of a polar functional group at the end of the 1-alkyl chain of these inactive pyrroles, however, restored or even elevated inhibitory potency. The most preferable of the polar terminal functions investigated was the carboxylic acid moiety. 6-[2-(2-Carboxyethyl)-4 dodecanoyl-3,5-dimethylpyrrol-1-yl]hexanoi c acid (65c) and 6-[2-(carboxymethyl) 4-dodecanoyl-3,5-dimethylpyrrol-1-yl]nonanoic acid (66f) were the synthesized inhibitors with the greatest potency. With IC50 values of 3.4 and 3.3 microM, respectively, they were about 3-fold more active than the standard cPLA2 inhibitor arachidonyl trifluoromethyl ketone (IC50: 11 microM). PMID- 9341914 TI - New orally active non-peptide fibrinogen receptor (GpIIb-IIIa) antagonists: identification of ethyl 3-[N-[4-[4-[amino[(ethoxycarbonyl) imino]methyl]phenyl] 1,3-thiazol-2-yl]-N-[1-[(ethoxycarbonyl)methyl]pip erid -4-yl]amino]propionate (SR 121787) as a potent and long-acting antithrombotic agent. AB - The platelet fibrinogen receptor GpIIb-IIIa is currently considered a target of choice for drugs used in the prevention and treatment of thrombosis. Ethyl 3-[N [4-[4-[amino[(ethoxycarbonyl)-imino] methyl]phenyl]-1,3-thiazol-2-yl]-N-[1 [(ethoxycarbonyl)methyl] piperid-4-yl] amino]propionate (6, SR 121787) is a new antiaggregating agent which generates in vivo the corresponding diacid 19d (SR 121566), non-peptide GpIIb-IIIa antagonist. In vitro, 19d inhibited ADP-induced aggregation of human and baboon platelets (IC50 = 46 +/- 11 and 54 +/- 6 nM, respectively), and on human platelets, 19d antagonized the binding of 125I labeled fibrinogen (IC50 = 19.2 +/- 6.2 nM). Ex vivo, 8 h after an i.v. administration of 19d (100 micrograms/kg, i.v.) to baboons, ADP-induced aggregation was strongly inhibited (more than 90%). At 8 h, the ED50 value was 24 +/- 3.3 micrograms/kg), and even 24 h after the administration of a single dose of 100 micrograms/kg of 19d, platelet aggregation was still significantly inhibited (50 +/- 6% inhibition, P < 0.05). In the same species, the oral administration of 500 micrograms/kg of 6 produced a nearly complete inhibition of aggregation for up to 8 h (ED50 at 8 h was 193 +/- 20 micrograms/kg). After an oral dose of 2 mg/kg of 6, an antiaggregating effect was still observed at 24 h (44 +/- 12% inhibition, P < 0.05). 6 was well tolerated in animals, showing that, on the basis of these studies, it is a suitable candidate for development as an orally active antithrombotic agent. PMID- 9341915 TI - Synthesis and stereochemical structure-activity relationships of 1,3 dioxoperhydropyrido[1,2-c]pyrimidine derivatives: potent and selective cholecystokinin-A receptor antagonists. AB - The synthesis and stereochemical structure--activity relationships of a new class of potent and selective non-peptide cholecystokinin-A (CCK-A) receptor antagonists based on the 1,3-dioxoperhydropyrido[1,2-c]pyrimidine skeleton are described. The most potent member of this series of eight diastereoisomers, (4aS,5R)-2-benzyl-5-[N-[(tert-butoxycarbonyl)-L-tryptophyl]-amino] - 1,3 dioxoperhydropyrido[1,2-c]pyrimidine, displays nanomolar CCK-A receptor affinity and higher than 8000-fold potency at the CCK-A than at the CCK-B receptor. As CCK A antagonist, this compound inhibits the CCK-8-evoked amylase release from pancreatic acinar cells at a low concentration, similar to that of the typical antagonist Devazepide. Highly strict stereochemical requirements for CCK-A receptor binding and selectivity have been found. The L-Trp and the 4a,5-trans disposition of the bicyclic perhydropyrido[1,2-c]pyrimidine are essential for binding potency and selectivity. PMID- 9341916 TI - Anionic- and lipophilic-mediated surface binding inhibitors of human leukocyte elastase. AB - We report the synthesis of a series of diphenylmethane-based oligomers containing anionic and lipophilic functionalities that are potent inhibitors of human leukocyte elastase (HLE). The enzyme inhibition is regulated by the size of the oligomer, as well as, the number of charges. Lipophilicity is an important element in determining potency and specificity against other basic enzymes. Compounds whose scaffolds contain three phenoxyacetic acid groups and three alkyl ethers are competitive and specific inhibitors of HLE with Ki = 20 nM. The mechanism of action of this class of compounds is believed to involve multidendate interactions with the surface of HLE near the active site which prevents substrate access to the catalytic site. PMID- 9341917 TI - 7-alkylidenecephalosporin esters as inhibitors of human leukocyte elastase. AB - A series of 7-alkylidenecephalosporins and 7-vinylidenecephalosporins, as their benzhydryl esters, have been tested as inhibitors of both porcine pancreatic elastase and human leukocyte elastase. Selected 7-alkylidenecephalosporin esters are found to be potent inhibitors of HLE. One category of new inhibitors is the 7 (haloalkylidene)cephalosporins. In contrast to previously reported cephalosporin based elastase inhibitors, these haloalkylidene cephems show optimum inhibitory activity as sulfides, rather than as sulfones. They are efficient and irreversible inhibitors. A second class of active compounds is represented by the benzhydryl ester 7-(cyanomethylidene)cephalosporin sulfone. In contrast to the activity of these new inhibitors, the benzhydryl ester of the mechanism-based beta-lactamase inhibitor, 7-[(2'-pyridyl)methylidene]-cephalosporin sulfone showed little inhibitory activity as an elastase inhibitor. 7 Vinylidenecephalosporins were also relatively poor inhibitors, although the terminally unsubstituted allene sulfide showed activity as an inhibitor of PPE. A modeling analysis suggests the 7-alkylidene substituents can be readily accommodated in the S1 pocket. A potential mechanism of inhibition is proposed. PMID- 9341918 TI - Cephalosporin 3'-phloroglucide esters and 7-(phloroglucidamido) cephalosporins as novel antibacterial agents. AB - Two series of new phloroglucide derivatives were synthesized that possessed antibacterial activities. The first series includes cephalosporin 3' phloroglucide esters 19 and 20, which were obtained by condensation of cephalosporin 16 with bioactive phloroglucides 14 and 15, respectively. They exhibited a dual mode of antibacterial action. In comparison with cephalosporins 26 and 27, bearing an acetoxy unit at the C-3' position, the bifunctional cephalosporins 19 and 20 showed a broadened spectrum of activity. Results from the consistent valence force field (CVFF) calculations indicate that the most stable conformational isomer of phenolic acid 14, holding a cis-syn-syn geometry, possessed a cavity. It provides an ideal environment to accommodate metal ions of holoenzymes. Phenolic keto acid 15, however, possessed a trans-anti-syn conformation, which allowed chelation between metal ions and the phenolic hydroxyl groups as well as the carbonyl functionalities. Our biological results show that the cavity formed in phloroglucides plays an important role. The second series includes 7-(phloroglucidamido)cephalosporins 24 and 25, which were synthesized by condensation of cephalosporin 21 with 14 and 15, respectively. Results from the CVFF calculations indicate that cephalosporin 24 also possessed a cavity. Unlike cephalosporin 3'-phloroglucide esters 19 and 20, cephalosporins 24 and 25 were found resistant to beta-lactamases from Staphylococcus aureus 95 and Pseudomonas aeruginosa 18S-H. These new compounds, however, showed notable activities against S. aureus FDA 209P, S. aureus 95, Candida albicans, P. aeruginosa 1101-75, and P. aeruginosa 18S-H. PMID- 9341919 TI - Thromboxane modulating agents. 3. 1H-imidazol-1-ylalkyl- and 3-pyridinylalkyl substituted 3-[2-[(arylsulfonyl)amino]ethyl]benzenepropanoic acid derivatives as dual thromboxane synthase inhibitor/thromboxane receptor antagonists. AB - The design of a series of dual thromboxane synthase inhibitor/thromboxane receptor antagonists based on a 3-[2-[(arylsulfonyl)amino]ethyl]benzenepropanoic acid thromboxane receptor antagonist template is described. Introduction of a 5 (1H-imidazol-1-ylmethyl), a 5-(3-pyridinyl-methyl), or a 5-(3-pyridinyloxy) substituent leads to dual agents with thromboxane synthase inhibitory activity comparable with that of dazmegrel (7). In addition, 3-pyridinylalkyl substituents also make a significant contribution to thromboxane receptor binding. Oral administration of compound 74 (5 mg/kg) to conscious dogs produces long-lasting thromboxane synthase inhibition and thromboxane receptor blockade as measured by inhibition of U46619-induced platelet aggregation ex vivo. PMID- 9341920 TI - Protein structure determination using a combination of comparative modeling and NMR spectroscopy. Application to the response regulator protein, Spo0F. AB - A practical combination of comparative modeling and NMR spectroscopy was used to generate a three-dimensional structure of the response regulator protein, Spo0F. The backbone structure obtained compares to the Spo0F Y13S mutant X-ray structure with an rmsd of 2.0 A. We provide results which suggest that structures obtained by this method are suitable for drug discovery. The results of the GRID and DOCK methods as applied to the model and X-ray structures of Spo0F are remarkably similar and tend to suggest the same design conclusions. This trend is illustrated by these same techniques applied to two experimentally derived structures of the analogous protein, CheY, which exhibit a pairwise rmsdBB on the same order as that found for the two Spo0F structures. PMID- 9341921 TI - Syntheses and biological activities (topoisomerase inhibition and antitumor and antimicrobial properties) of rebeccamycin analogues bearing modified sugar moieties and substituted on the imide nitrogen with a methyl group. AB - As a part of studies on structure-activity relationships, several potential topoisomerase I inhibitors were prepared. Different analogues of the antitumor antibiotic rebeccamycin substituted on the imide nitrogen with a methyl group were synthesized. These compounds bore either the sugar residue of rebeccamycin, with or without the chlorine atoms on the indole moieties, or modified sugar residues (galactopyranosyl, glucopyranosyl, or fucopyranosyl) linked to the aglycone via a beta- or alpha-N-glycosidic bond. Their inhibitory properties toward protein kinase C, topoisomerase I, and topoisomerase II were examined, and their DNA-binding properties were investigated. Their in vitro antitumor activities against murine B16 melanoma and P388 leukemia cells were determined. Their antimicrobial activities were tested against Gram-positive bacteria Bacillus cereus and Streptomyces chartreusis, Gram-negative bacterium Escherichia coli, and yeast Candida albicans. These compounds are inactive toward topoisomerase II but inhibit topoisomerase I. A substitution with a methyl group on the imide nitrogen led to a loss of proteine kinase C inhibition in the maleimide indolocarbazole series but did not prevent topoisomerase I inhibition. Compounds possessing a beta-N-glycosidic bond, which fully intercalated into DNA, were more efficient at inhibiting topoisomerase I than their analogues with an alpha-N-glycosidic bond; however, both were equally toxic toward P388 leukemia cells. Dechlorinated rebeccamycin possessing a methyl group on the imide nitrogen was about 10 times more efficient in terms of cytotoxicity and inhibition of topoisomerase I than the natural metabolite. PMID- 9341922 TI - 19-nor-10-azasteroids, a new class of steroid 5 alpha-reductase inhibitors. 2. X ray structure, molecular modeling, conformational analysis of 19-nor-10 azasteroids and comparison with 4-azasteroids and 6-azasteroids. AB - 19-Nor-10-azasteroids are a new class of 5 alpha-reductase inhibitors whose activity depends on the presence of the bridgehead N-10 atom conjugated with the 4-en-3-one moiety in the A ring. The X-ray structure of 19-nor-10-azasteroid 1 has been determined and it is compared with the X-ray structure of testosterone. A complete conformational analysis of these compounds has been performed, determining the number and energy of the possible conformers, as well as the molecular flexibility of the 10-azasteroidal skeleton. Thus, MM2* molecular mechanics calculations and AM1 semiempirical energy refinements revealed that 19 nor-10-azasteroids 1-3 have four possible conformations with very small energy differences and that they are very flexible molecules. The conformational analysis has been extended to testosterone (4), which also showed conformational flexibility, with three different conformations, and to 6-azasteroid 5 and 4 azasteroid 6, for which only two thermally accessible conformations have been found. Compared to 19-nor-10-azasteroids 1-3, azasteroids 5 and 6 appear to be more rigid structures. By a best fit analysis of all conformers of 1-5 with the global minimum of testosterone (4-I) it has been found that the lowest energy conformers of 1, 3, and 5 are very close to the structure of 4-I, and among the conformers of 2, the best similarity has been observed for the highest energy conformer 2-IV. PMID- 9341923 TI - Synthesis and antiviral activity of C2 analogs of enviroxime: an exploration of the role of critical functionality. AB - Enviroxime is a potent antiviral agent with broad spectrum activity in tissue culture against both rhinoviruses and enteroviruses. We have synthesized and studied a series of C2-substituted analogs in order to identify critical functionality and examine its role in antiviral activity. We have found that primary amino substitution is the most active. Ab initio calculations indicate that larger groups at C2 may provide a repulsive steric interaction at N3, and in those cases where this undesirable conformation has limited flexibility, the antiviral activity is severely reduced. Further the results show that an amino hydrogen at C2 is strongly hydrogen bonded to the N1 sulfonyl oxygen, which in the case of Enviroxime may act to enhance the activity by holding the second hydrogen in a desirable orientation for interaction at an antiviral site. PMID- 9341925 TI - Prenatal US detection of cerebral ventricular enlargement in a case of tuberous sclerosis. PMID- 9341924 TI - Synthesis, cardiotonic activity, and structure-activity relationships of 17 beta guanylhydrazone derivatives of 5 beta-androstane-3 beta, 14 beta-diol acting on the Na+,K(+)-ATPase receptor. AB - A series of digitalis-like compounds, with the lactone ring shifted from the original position through a spacer or replaced by a series of guanylhydrazone substituent-bearing chains, was synthesized and evaluated for inhibition of Na+,K(+)-ATPase and for inotropic activity. The highest Na+,K(+)-ATPase inhibition (IC50) and inotropic activity (EC50) were reached with the vinylogous guanylhydrazone 5 where a cardenolide-like polarized alpha,beta-unsaturated system and a basic guanidino group were both present at the 17 beta-position; for this compound IC50 and EC50 values were comparable to or higher than those of Thomas' parent guanylhydrazone 1, digitoxigenin, and digoxin. A substantial improvement of the desired positive inotropic activity versus the toxic arrhythmogenic concentration was not reached within this series; only a slightly better therapeutic index can be envisaged for compounds 5 and 4, even though, for the latter, to the detriment of potency, presumably because of a weaker interaction with the receptor, due to the lack of a cardenolide-like polarized system. PMID- 9341926 TI - Thermal therapy of functional dyskinesias of the alimentary tract. AB - There are described the studies performed in Italy-especially at Montecatini about activity of certain mineral waters, administered by mouth, in functional troubles of the digestive apparatus. The illnesses especially considered have been: idiopathic chronic constipation, irritable colon syndrome, biliary dyskinesias, correlated pathological conditions. The mineral waters analyzed had been: salso-sulphate-alkaline, bicarbonate, sulphate, bicarbonate-sulphate alkaline, sulphate-bicarbonate waters and others. There are reported: research methods employed, the obtained results, the possible mechanisms of effect. The hydrological favourable influences about pathophysiology of digestive motor activity and in therapeutics of correlated diseases are demonstrated. The clinical results, and particularly their duration in time, are affermatif about therapeutic usefulness of thermal treatment by mouth. It's possible to suggest and to stress their better knowledge in Medicine and their increased employment in the treatment of functional dyskinesias of the alimentary tract. PMID- 9341929 TI - Alterations of cell cycle regulatory genes in primary (de novo) and secondary glioblastomas. AB - Primary glioblastomas develop rapidly de novo through a genetic pathway characterized by amplification/overexpression of EGFR and of MDM2 genes. Secondary glioblastomas develop more slowly through progression from low grade or anaplastic astrocytoma and show a high incidence of a p53 mutation. In the present study, primary and secondary glioblastomas were analyzed for p16 deletions and CDK4 amplification by differential PCR and for loss of expression of the retinoblastoma (RB) gene by immunohistochemistry. Except for one case, alterations in the structure or expression of p16, CDK4 and RB were mutually exclusive. The overall incidence of aberrant expression of these genes coding for components of the cell-cycling-regulatory system was similar in primary (14/28; 50%) and secondary glioblastomas (9/23; 39%). However, p16 deletions were significantly more frequent in the former (10/28; 36%) than in the latter (1/23, 4%; P = 0.0075), suggesting that this alteration constitutes an additional genetic hallmark of the primary (de novo) glioblastoma. PMID- 9341930 TI - Schwann cell differentiation in Charcot-Marie-Tooth disease type 1A (CMT1A): normal number of myelinating Schwann cells in young CMT1A patients and neural cell adhesion molecule expression in onion bulbs. AB - Charcot-Marie-Tooth disease type 1A (CMT1A) is a common hereditary demyelinating neuropathy caused by a duplication of the gene for the myelin protein PMP22, resulting in overexpression of PMP22 in young patients. Although genetically well defined, the pathogenesis of the hereditary demyelinating neuropathy CMT1A is still unclear. Homology of PMP22 cDNA to the growth arrest-specific gene gas3 and experiments in vitro showing decreased proliferation in PMP22-overexpressing Schwann cells suggest a role of PMP22 in Schwann cell differentiation. Furthermore, overexpression of PMP22 in fibroblasts induces programmed cell death. In this report we applied morphometrical methods using electron micrographs and immunohistochemistry to further characterise Schwann cells in CMT1A nerve biopsy samples from CMT1A patients. We show that the total number of PMP22-expressing Schwann cells, i.e. Schwann cells that are in a 1:1 relationship with axons, was not reduced in sural nerve biopsy samples from six young CMT1A patients. We excluded non-specific secondary Schwann cell proliferation. Thus, in young CMT1A patients with increased PMP22 overexpression there seems to be no evidence for altered initial Schwann cell proliferation in achieving a 1:1 relationship to axons prior to the process of de- and remyelination. Further, using electron microscopy we found no evidence for apoptosis of Schwann cells in CMT1A. However, we provide additional support for an abnormal Schwann cell phenotype in CMT1A by showing the expression of neural cell adhesion molecule immunoreactivity in onion bulbs. Thus, the role of PMP22 in cell growth and differentiation does not lead to an altered number of myelinating Schwann cells but to altered Schwann cell differentiation in CMT1A. PMID- 9341931 TI - Microglial activation in early stages of amyloid beta protein deposition. AB - The present study was undertaken to investigate the relationship of microglial activation to amyloid beta protein (A beta) deposition, particularly at the early stage. Using single and double immunostaining methods with a panel of microglia markers and antibodies against A beta and amyloid beta protein precursor (APP), we examined the cerebrum and cerebella of both Alzheimer's disease (AD) and non demented subjects obtained at autopsy. In nondemented, middle-aged subjects that had small amounts of cerebral A beta deposits, approximately 70% of the diffuse plaques contained ramified microglia. However, no evidence of microglial activation was found in diffuse plaques in any of the non-demented subjects. Dual immunostaining of sections of cerebral cortex using antibodies against A beta and major histocompatibility complex class II antigen showed that in AD subjects, approximately 20% of total diffuse plaques contained a few, activated microglia. Most of these plaques were defined as a transitional from between diffuse and primitive plaques. Both primitive and classic plaques in the cerebral cortex of AD subjects consistently contained clusters of activated microglia. Subpial A beta deposits without neuritic changes lacked microglial activation. In the cerebellum, all of the diffuse plaques lacked microglial activation, and activated microglia in the compact plaques were not as hypertrophic as those in cerebral primitive/classic plaques. Our findings indicate that microglial reactions are absent in the early stages of A beta deposition, and it occurs during the transition from diffuse to primitive plaques, when amounts of A beta deposits and the degree of neuritic changes increase. PMID- 9341932 TI - Deposition of amyloid beta protein (A beta) subtypes [A beta 40 and A beta 42(43)] in canine senile plaques and cerebral amyloid angiopathy. AB - To clarify the immunohistochemical features of canine senile plaques (SPs) and cerebral amyloid angiopathy (CAA), the distribution of the amyloid beta protein (A beta) subtypes A beta 40 and A beta 42(43), A beta precursor protein (APP), and glial cell reaction were examined in the brains of seven aged dogs (12-18 years). A beta 42(43) was found to be deposited in all types of SPs, whereas A beta 40 was deposited only in mature (classical and primitive) plaques. CAA, which was located along parenchymal and meningeal arterioles and capillaries, consisted of both subtypes of A beta. APP was exhibited in normal and degenerative neurons and swollen neurites of mature plaques. It was, therefore, considered that A beta 42(43) in diffuse plaques might be derived from APP in neurons, while A beta 40 and A beta 42(43) in mature plaques might be generated from APP in swollen neurites in the plaque. In contrast to the case in humans, in whom deposition of A beta 40 and A beta 42(43) in the mature plaques is predominantly associated with microglial reaction, in dogs we found that it was closely associated with astroglial reaction. The present findings showed characteristics of canine SPs which are different from those of humans. PMID- 9341933 TI - Characterization of a prolonged regenerative attempt by diffusely injured axons following traumatic brain injury in adult cat: a light and electron microscopic immunocytochemical study. AB - Traumatic brain injury in animals and humans is well known to cause axonal damage diffusely scattered throughout the brain without evidence of other brain parenchymal change. This observation has prompted some to posit that such damaged axons are well positioned to mount a regenerative attempt. The present study uses an immunocytochemical marker specific for regenerating neurites to explore this issue. Further, in an attempt to expedite and enhance any potential regenerative effort, this study evaluates the efficacy of intrathecally applied nerve growth factor. Three sets of experiments were performed in adult cats. One group of animals was subjected to moderate fluid percussion brain injury and followed for 7 or 14 days post injury, with the continuous intraventricular infusion of nerve growth factor delivered by implanted osmotic pumps. These animals were compared to a second group of time-matched, sham-operated animals receiving artificial cerebrospinal fluid infusion. To assess axonal damage immunohistochemical staining for the low molecular weight neurofilament subunit (NF-L) was carried out using an NR4 monoclonal antibody. To localize axons exhibiting a regenerative response immunohistochemical staining for the growth associated protein GAP43 was employed. In sham controls, at the light microscopic level NF-L-immunoreactive axonal swellings were numerous at 7 days, but by 14 days post injury their frequency declined markedly. In contrast, GAP43-immunoreactive, disconnected reactive axonal swellings were rarely observed at 7 days but were numerous at 14 days. Ultrastructural analysis at 14 days post injury of carefully matched sections revealed reactive axons demonstrating sprouting consistent with a regenerative effort. Analysis of tissue from animals of 14 days of survival indicated that supplementation with nerve growth factor did not appear to enhance the capacity of damaged brain axons to mount a regenerative attempt. Rather, it appears that regenerative efforts seen reflect a spontaneous response. A third group of adult cats, subjected to the same injury but not subjected to osmotic pump implantation, was allowed to survive for 22-28 days. Animals in this group also demonstrated GAP43 immunoreactivity in reactive axonal swellings in the brain stem. This study demonstrates that diffusely injured axons can mount a sustained regenerative attempt that is associated with a reorganization of their cytoskeleton and accompanied by an up-regulation of growth-associated proteins. PMID- 9341934 TI - Immunocytochemical and ultrastructural study of pericapillary rosettes in amyotrophic lateral sclerosis. AB - This report concerns a comparative immunocytochemical and ultrastructural investigation on pericapillary rosettes (PR) in the lumbar spinal cords of 21 patients with amyotrophic lateral sclerosis (ALS) and 18 age-matched neurologically normal individuals. The purpose of the study was to determine the alteration of PR in relation to the neuronal loss in ALS. The PR were almost always positively immunostained for phosphorylated neurofilament, and some PR immunoreacted with antibodies to synaptophysin and beta-amyloid precursor protein. This finding suggests that axonal transport, whether fast or slow, is impaired in the terminal portion of the axon that reaches the capillaries. Some PR were also positively immunostained by the antibody against ubiquitin, anti calbindin-D 28 K antibody, anti-parvalbumin antibody and the antibody to superoxide dismutase 1. Morphometrically, the number of PR in the anterior horns and lateral column was markedly diminished in ALS compared with controls. At the ultrastructural level, the PR consisted mostly of unmyelinated degenerated axons, and were frequently found outside the basal laminae of the endothelial cell and of the astrocytic foot processes on the opposite side of the capillary, and less often in the space between the two basal laminae. The data indicate that the fate of PR is intimately associated with the neuronal loss of the anterior horn cells and with degenerative change of nerve fibers extending from their mother neurons to the capillaries. PMID- 9341935 TI - Changes in weight and compositions of major membrane components of human brain during the span of adult human life of Swedes. AB - Brain weight, total solids, protein, and major lipids have been determined in 83 female and 101 male brains from subjects 20-100 years of age. The brain weight began to diminish at 20 years of age. The brain weight at 20 years for females: 1,368 +/- 26 and for males 1,632 +/- 27 g diminished at 100 years for females to 1,100 +/- 25 and for males to 1,266 +/- 25 g, a decrease of 20% for female and 22% for male brains. The decrease in dry solids was larger during the same period, 36% for females and males. Proteins decreased by 39% in females and 37% in males. Phospholipids decreased by 42% in females and 43% in males, cholesterol by 47% and 53%, cerebroside by 46% and 58%, sulfatide by 46% and 49% and gangliosides by 28% and 30%, respectively. There is, thus, a significantly larger loss of myelin lipids than of gangliosides-the biochemical marker for neuronal membranes. The loss of myelin lipids was particularly large in female brain after 70 years of age, while the loss in male brain was linear as early as from 20 years of age. PMID- 9341936 TI - Argyrophilic grain disease: distribution of grains in patients with and without dementia. AB - In a previous study we reported on a late onset dementia which occurred in only half of the patients with argyrophilic grain disease (AgD) investigated. To find a correlation between the distribution of argyrophilic grains (ArG) and the occurrence of a late onset dementia, we examined the limbic area in 35 subjects who had ArG as the main neuropathological finding. A retrospective clinical analysis was performed by collecting information from hospital charts supplemented by standardized interviews based on DSM IV criteria for dementia. Sections from the rostral and caudal hippocampal regions, including the entorhinal/transentorhinal and parahippocampal cortex on both sides, were strained by the Gallays method. Nineteen subjects were diagnosed as demented according to these criteria; 16 were considered to have been cognitively normal. High numbers of ArG were observed in the anterior part of the CA1 subfield in all cases. However, the posterior half of CA1 was involved significantly more often and more severely in demented than in non-demented individuals (P < 0.01). Moreover, the distribution of ArG in the entorhinal/transentorhinal and parahippocampal cortex was more widespread in the group of demented patients (P < 0.05). These results show that the intellectual status of patients with AgD was related to the extension of ArG in the limbic area. We suggest that AgD is a progressive neurodegenerative disorder with early subclincial lesions in the anterior part of the hippocampal formation. To provide a more accurate clinicopathological correlation, the rostrocaudal extension of ArG in the limbic area should be evaluated in AgD cases. PMID- 9341937 TI - The monocyte-macrophage system is affected in lysosomal storage diseases: an immunoelectron microscopic study. AB - Studying peripheral blood mononuclear cells (PBMCs) has become an important diagnostic tool in lysosomal storage diseases. Previous studies revealed that B and subclasses of T lymphocytes participate in the storage process, whereas the role of circulating monocytes was not clear. In this study, the involvement of CD14+ monocytes in lysosomal diseases was investigated. Blood samples from six patients with different lysosomal storage disorders were studied, including one with late--infantile and three with juvenile neuronal ceroid--lipofuscinoses, and two with mucopolysaccharidosis type VI. CD14+ cells were separated immunomagnetically from PBMCs and studied by light and electron microscopy. In all investigated disorders, disease-specific lysosomal storage material could be found in monocytes. The ratio of affected to non-affected cells did not differ from previously reported data on lymphocytes and their subforms in these diseases. Our data were obtained by studying a small number of different lysosomal storage disorders. Nevertheless, they suggest that lysosomal storage in the monocyte-macrophage system might also be found in other forms of lysosomal diseases. PMID- 9341938 TI - Oral coenzyme Q10 administration prevents the development of ischemic brain lesions in a rabbit model of symptomatic vasospasm. AB - Treatment with oral coenzyme Q10 (CoQ10, 10 mg/kg per day for 6 days) was compared with no treatment in a previously described rabbit model of symptomatic cerebral vasospasm [Endo et al. (1988) Stroke 19: 1420-1425]. The treatment was initiated within 1-2 h after injection of autologous blood into the subarachnoid space. In CoQ10-untreated rabbits, moderate to severe neurological deficits developed, and multiple focal ischemic lesions were found in the brain regions with compromised blood supply, i.e., in the regions normally supplied by common carotid arteries which are subject to ligation in this model. CoQ10 treatment prevented the development of both the neurological deficits and histologically detectable brain tissue damage. In both CoQ10-treated and -untreated rabbits, infiltration of mononuclear cells was evident in the brain stem, although this region did not show signs of ischemic damage. The findings indicate that the histological and neurological correlates of brain tissue damage in this rabbit model of symptomatic cerebral vasospasm develop via mechanism(s) involving free radical-mediated oxidation of plasma lipoproteins. Similar mechanisms may play a role in the development of brain damage attributed to cerebral atherosclerosis. PMID- 9341939 TI - bcl-2 protein expression in astrocytomas in relation to patient survival and p53 gene status. AB - bcl-2 protein expression was characterized in a series of 58 astrocytomas from 21 pediatric and 37 adult patients. As part of a continuing attempt to define relevant prognostic factors which may predict clinical outcome, we have determined the impact of bcl-2 accumulation in malignant astrocytes on the length of patient survival. Aberrant overexpression of bcl-2 protein in tumor cells was detected in 57% (12 of 21) of pediatric and 73% (27 of 37) of the adult cases. Among pediatric patients, the median survival in months showed no relationship with the incidence of bcl-2-positive tumors. Among the adult patients, a favorable prognostic indicator was low-tumor grade (P = 0.05). bcl-2-positive tumors occurred with similar frequencies in WHO grades III and IV of malignancy. When bcl-2 expression in tumor cells was tested as a variable to predict for patient survival, the 6 patients without bcl-2 expression among 23 adult patients with grade IV tumors had a shorter median survival. The same 58 tumors had been previously analyzed for alterations of p53:4 pediatric and 16 adult tumors had p53 gene mutations. There was no significant difference in median survival related to p53 gene status. There was no relationship between bcl-2 expression and p53 gene status: approximately equal numbers of tumors with either wild-type or mutant p53 were bcl-2 negative or bcl-2 positive. bcl-2 expression is high (40 100%) among other tumors of the central nervous system which also show low malignant potential. Up-regulation of bcl-2 in malignant astrocytes or constitutive expression in some tumor types may be a factor leading to a more favorable clinical outcome. PMID- 9341940 TI - Glial fibrillary acidic protein expression in a new human glioma cell line in culture before and after xenogenic transplantation into nude mice. AB - A human glioma cell line, SA146, was initiated on precoated extracellular matrix from a stereotactic biopsy of a glioblastoma. We report modulation in the expression of glial fibrillary acidic protein (GFAP) by SA146 passed in vitro before or after xenogenic transplantation into nude mice. Immunofluorescence data show a decrease in the percentage of GFAP-expressing cells with increasing in vitro passages but a full reexpression (100% of GFAP-positive cells among vimentin-positive cells) was observed in cultures just derived from the xenotransplanted tumor. These changes are correlated with the mRNA content (Northern blot probed with a cDNA for GFAP) and with the protein level (cytoskeletal fraction analyzed by two-dimensional gel electrophoresis and Western blots probed with a monoclonal antibody). At the optimal level of GFAP expression, a large range of micro-heterogeneity in GFAP isoforms is reached for which post-translational events are clearly involved since mRNA translation in cell free system would provide at best three isomers. We suggest that SA146 would be an appropriate model to study the regulation of GFAP expression in the context of human glial tumor biology. PMID- 9341941 TI - Idiopathic hypertrophic cranial pachymeningitis mimicking multiple meningiomas: case report and review of the literature. AB - A case of idiopathic hypertrophic cranial pachymeningitis with an unusual and misleading manifestation is reported. Computed tomography scan, angiographic and magnetic resonance imaging findings were suggestive of multiple meningeal neoplasms and a correct diagnosis was made only after meningeal biopsy. This 44 year-old patient had a previous history of an ill-defined systemic disorder associating episcleritis, erythroderma nodosa and multiple peripheral arthritis. We review previous reports of idiopathic cranial pachymeningitis with emphasis on radiological investigation techniques, histopathology and possible dysimmune mechanisms of pathogenesis. PMID- 9341942 TI - A canine encephalomyelopathy with morphological abnormalities in mitochondria. AB - A progressive encephalomyelopathy of insidious onset affecting a 16-month-old dog is described. Clinically, the dog was ataxic, stumbled into objects and showed mild behavioral abnormalities. Light microscopic findings included profound degeneration and astrogliosis of the optic pathways, loss of Purkinje neurons, focal bilateral and symmetrical brain stem spongiosis and diffuse neuroaxial astrogliosis with swollen and abnormally shaped nuclei. Ultrastructurally, there were giant and bizarre mitochondria within neuronal perikarya and axons as well as diffuse loosening of the cerebral and cerebellar neuropil. These neuropathological findings resemble the mitochondrial encephalomyopathies of man. PMID- 9341943 TI - Primary cerebellar extramedullary myeloid cell tumor mimicking oligodendroglioma. AB - Extramedullary myeloid cell tumors (EMCTs) are tumors consisting of immature cells of the myeloid series that occur outside the bone marrow. Most of them are associated with acute myelogenous leukemia or other myeloproliferative disorders, and a small number occur as primary lesions, i.e., are not associated with hematological disorders. Occurrence inside the cranium is rare, and there has been only one case of primary EMCT involving the cerebellum reported in the literature. The case we report here is a blastic EMCT occurring in the cerebellum of a 3-year-old boy who had no signs of leukemia or any hematological disorder throughout the entire course. The cerebellar tumor was at first misdiagnosed as an "oligodendroglioma" because of the uniformity and "fried egg" artifact of the tumor cells. The tumor disappeared during chemotherapy consisting of 12 treatments. However, it recurred and metastasized to the cerebrospinal fluid (CSF) shortly after the therapy was completed. A diagnosis of EMCT was suspected because of the presence of immature myeloid cells in the CSF, and was confirmed by anti-myeloperoxidase and anti-lysozyme immunoreactivity of the cerebellar tumor. The patient succumbed 1 year and 3 months after the first presentation of the disease. PMID- 9341944 TI - 3rd CPA-Symposium: Neuromonitoring and Cerebral Protection. Homburg-Saar, November 1995. PMID- 9341945 TI - Angioarchitectural classification of the fungiform papillae on the dorsal surface of the bullfrog tongue. AB - In this study, the three-dimensional anatomy of the microvascular structure of fungiform papillae (FuP) on the dorsal surface of the bullfrog tongue has been investigated using scanning electron microscopy (SEM), and angioarchitectural classification has been carried out by means of the capillary loop (L. s) and intra-bridge structure (I. b). FuP from 30 sound bullfrog tongues were used. The following research methods were applied: SEM observation on microvascular cast specimens (MVCS) of bullfrog tongue's FuP were injected with synthetic resin (Mercox). Observation of MVSC showed that the bullfrog tongue FuP consisted of an ascending branch (A. b), L. s, I. b and a descending branch (D. b). Based upon SEM observations of MVCS, FuP can be classified into four types: A, B, C, D types according to A. b, L. s, I. b and D. b. A-type (none I. b) formed from A. b, L. s, D. b only and with no I. b. B-type (one I. b) consisted of A. b, L. s, D. b and one I. b. C-type (two I. b) were composed of A. b, L. s, D. b and two I. b. D type (three I. b) were composed of A. b, L. s, D. b and three I. b, A. b, L. s, I. b and D. b on each. A, B, C, D types were all same thickness. PMID- 9341946 TI - Angioarchitectural structure of the fungiform papillae on the anterodorsal surface of the rat tongue. AB - To ascertain whether the microvascular morphological differences of fungiform papillae (FuP) on the anterodorsal surface of the rat tongue are locationally and functionally related, this study aimed at examining and comparing, in greater detail, the comparative morphological characteristics of FuP. FuP were sporadically and consistently scattered among numerous filiform papillae (FiP) in three parts: the apical, central and in front of the intermolar eminence on the anterodorsal surface. We studied these by means of the microvascular cast specimen (MVCS) of FuP using scanning electron microscopy (SEM). The results obtained in this study showed that in all three parts on the anterodorsal region, the capillary bed of FuP presented as a netbasket-like and cylindrical figure with a central hole consisting of 4-5 horizontal rings of several ascending and descending roots. FuP in the central part on the anterodorsal region were relatively larger in size and more cylindrical in shape than those of both the other parts, and they play the leading part as sensory organs for the taste sense and were regarded as taste receptive organs. PMID- 9341947 TI - A quantitative backscattered electron imaging study of hypomineralization and hypoplasia in fluorosed dental enamel of deer. AB - Mineral content and distribution of fluorosed and unfluorosed (control) dental enamel of roe deer and red deer cheek teeth were analyzed using digital backscattered electron (BSE) imaging of PMMA-embedded specimens. Compared to the controls, the fluorosed enamel exhibited various aberrations resulting from a fluoride-induced disturbance of the processes involved in enamel formation. Thus, the presence of surface hypoplasias and an enhancement of the incremental pattern in the fluorosed enamel are evidence of a fluoride impact on the secretory ameloblasts, whereas a (subsurface) hypomineralization of different depth and extent is indicative of a fluoride effect on the maturation stage of amelogenesis. The marked variation in the severity of enamel hypomineralization seen along the coronocervical axis of a specimen pointed to a fluoride impact of varying intensity during this period of tooth development. Our observations further indicated that, in some locations, ameloblasts severely affected by fluoride during enamel matrix formation were able to recover from this insult and to function quite normally during the maturation stage of amelogenesis. A major advantage of the BSE imaging technique used in the present study over other methods is that it allows for a combination of micromorphological information with quantitative data on the mineralization of the analyzed tissue, which proved to be very useful for the characterization of fluoride-induced changes in dental enamel. PMID- 9341948 TI - Proliferation and apoptosis in follicles of the marmoset monkey (Callithrix jacchus) ovary. AB - Proliferation and apoptosis were studied in ovarian follicles of immature and pubertal marmosets and in mature marmosets during the follicular, periovulatory and luteal phases. Proliferation was evaluated using a Ki 67 antibody and apoptosis was assessed by in situ detection of DNA fragmentation. In the immature animals only small follicles were present, and the expression of Ki 67 was restricted to the granulosa cells of follicles localised near the medulla. There was no evidence of DNA fragmentation. In pubertal and adult animals Ki 67 expression was found in the granulosa cells of some but not all primordial and primary follicles. In the secondary and tertiary follicles immunoreactivity was localized in theca cells and granulosa cells. In atretic follicles (morphologically classified) the number of Ki 67 positive granulosa cells varied. In corpora lutea as well as in corpora lutea accessoria, staining was seen in the nuclei of some luteal cells. During all phases of the cycle, follicles from the secondary stage onwards were proliferating, whereas granulosa cells of primary follicles were only stained during the follicular phase. During all phases of the ovarian cycle apoptosis was restricted to the granulosa cells of tertiary follicles. With regard to proliferation and apoptosis, follicles exhibiting morphological signs of atresia can be classified as follows: (1) granulosa cells showing strong Ki 67 expression; (2) granulosa cells with reduced expression of Ki 67; (3) granulosa cells devoid of Ki 67 immunoreactivity and of apoptotic signs; (4) granulosa cells heavily stained for DNA fragmentation and not stained for Ki 67; (5) granulosa cells close to the antrum showing DNA fragmentation but luteinizing Ki 67 positive granulosa cells close to the basement membrane. In summary, it was shown that atresia of tertiary follicles is characterised by three consecutive stages: morphological alterations, cessation of proliferation and finally apoptosis in tertiary follicles. Thus, our results indicate that early atresia as evidenced by the morphological signs is not necessarily related to DNA fragmentation, since apoptosis is exclusively found in the granulosa cells of advanced atretic tertiary follicles. PMID- 9341949 TI - Potassium bromide and the thyroid gland of the rat: morphology and immunohistochemistry, RIA and INAA analysis. AB - The increasing environmental concentration of bromine has resulted in attempts to obtain information on its possibly deleterious effect on humans, particularly on a major target organ of this halogen i.e. the thyroid gland. In order to establish the morphological and functional effects of bromine on the thyroid, we have performed experiments on male rats which, in addition to a standard diet with an estimated iodine/bromine content, were fed for periods of 16 and 66 days with the small quantities of bromide expected to be encountered in the environment (10, 50 and 100 mg of Br-/l in drinking water). This treatment induced growth of the follicular epithelial component and microfollicular tissue rearrangement, a reduction of intrafollicular colloid, an increase in the height of the follicular cells and the number of mitoses, and it enhanced vascularization. Image analysis revealed a significant reduction in the volume of colloid, despite the accompanying rise in the number of minute follicles. The immunohistochemical positivity of the thyroglobulin fell in the microfollicular colloid of the exposed animals, although this was affected to a lesser extent in the larger follicles. The concentration of bromine in the thyroid increased with the amount of bromine intake, while at the same time the molar ratio of iodine/bromine decreased. The plasma level of T4 was lowered after both 16 and 66 days of treatment, but the T3 level only after 66 days treatment. The level of TSH did not exhibit any significant change. The observed changes, which have a parenchymatous goitre-like character, may have a direct relevance for human medicine, since the concentrations of bromide chosen in these experiments are readily encountered in the environment. PMID- 9341950 TI - Extracellular matrix composition of different regions of the knee joint cartilage in cattle. AB - Articular cartilage covering the bone ends at the joint shows different chemical composition in different regions, depending on the mechanical and biological properties of that region. Several studies have shown a relationship between the chemical composition of the cartilage and biomechanical forces. In the present study we analysed five different knee joints divided into the following regions: F1-medial and lateral border of the patellar surface, F2-patellar surface of the femur, F3-medial and lateral condyles, P-articular surface of the patella and T medial and lateral condyle of the tibia. The main glycosaminoglycan (GAG) present in these regions was chondroitin sulfate. Analysis of total GAG after digestion of the tissue with papain showed that in F2 and F3 there was a larger quantity of GAG/mg tissue, probably due to the dynamic character of the biomechanical forces in these regions. No significant differences were found for the extract and D1 fractions of the different regions. Analysis of the D4 fraction showed that the protein content was higher in the F3 and P regions than in their opposite T and F2 regions. The differences among the five regions may be a result of the non uniform presence of biomechanical forces supported by these regions. It is important to consider that the intensity and direction of stress in different parts of a tissue may influence the composition of the extracellular matrix. PMID- 9341951 TI - Articular chondrocytes and synoviocytes in culture: influence of antioxidants on lipid peroxidation and proliferation. AB - Chondrocytes and synoviocytes are the main cell types in articular joints. Articular cartilage is fed by synoviocytes via synovial fluid and has a low partial oxygen pressure. Thus, chondrocytes show oxygen radical protective mechanisms in vivo and are unprotected against these factors under common culture conditions. We investigated the influence of ascorbic acid, Fe2+, glutathione and alpha-tocopherol on lipid peroxidation and proliferation of rat articular chondrocytes and rabbit synoviocytes (HIG-82) in vitro. A combination of ascorbic acid and Fe2+ induced the production of thiobarbituric acid-reactive material as a marker of radical-mediated lipid peroxidation in homogenates and/or supernatants of cultured chondrocytes and synoviocytes. The amount of lipid peroxidation of chondrocytes was about 3-fold higher than that of synoviocytes. Ascorbic acid or Fe2+ alone had no significant influence on the production of thiobarbituric acid-reactive material. Lipid peroxidation could be abolished by addition of the radical scavenger alpha-tocopherol, whereas glutathione had no effect. 25-50 microM alpha-tocopherol decreased the ascorbic acid-(100 micrograms/ml) and Fe(2+)-(3 microM) induced lipid peroxidation to a basal level. Moreover, ascorbic acid inhibited the proliferation of rat chondrocytes and rabbit synoviocytes measured by [3H]-thymidine incorporation. Alpha-tocopherol and glutathione had no influence on the proliferation of chondrocytes but alpha tocopherol decreased the growth of synoviocytes and increased the anti proliferative effect of ascorbic acid on these cells. The importance of these findings for the use of ascorbic acid, glutathione and alpha-tocopherol in chondrocyte and synoviocyte cultures, or the influence of these molecules on the etiology and treatment of articular diseases will be discussed. PMID- 9341952 TI - Organization of collagen and elastic fibers studied in stretch preparations of whole mounts of human visceral pleura. AB - Fibers of the collagenous and elastic systems are most relevant in the double mechanical action of visceral pleura (VP), i.e. volume limitation and the generation of elastic recoil pressure. In this work we studied the organization of these fibrous components of VP in two situations: normal lungs and bullous disease. We employed histochemical methods on conventional histological slides and on thin spreads of whole mounts of visceral pleura. In addition, the scanning electron microscope was also used. According to our results, pleural function is made possible by the combination of both the elastic and collagenous fiber systems, each one having as intrinsic organizational pattern. Marked alterations of pleural bullous structure are observed with changes in lung volume. Fibers of the elastic and collagenous systems are clearly interdependent elements. Collagenous fibers are interwoven in a plaited structure that closely resembles the osiers of a wicker basket, indicating that collagen fibers allow for lung volume increase up to a point of maximal stretching of the system. The pleural contribution to lung elastic recoil pressure originates from the elastic network which turns back to its resting position when inspiratory pressures are negligible. The pleural immobility in bullous disease is associated with an almost complete absence of elastic fibers and the presence of very thick collagen fibers, suggestive of a cicatricial process, devoid of any characteristic pattern of distribution. PMID- 9341953 TI - Diversity of pituitary cells in primary cell culture. An immunocytochemical study. AB - In cell cultures of dispersed rat anterior pituitary, the specific identification of each cell type based on their staining properties and the ultrastructural features of secretory granules has proved to be unreliable. The existence of pituitary cell subtypes and the striking remodeling of the cell surface and intracellular organelles, further complicate the specific identification of pituitary cell populations. An immunocytochemical study of dissociated pituitary cells in culture was carried out to identify the cellular hormonal content by applying specific antibodies against prolactin (PRL), and growth (GH), luteinizing (LH beta), adrenocorticotrophic (ACTH) and thyrotrophic (TSH) hormones. Specifically bound IgG was exposed by the electron microscope with protein A-gold complex. Typical lactotrophs, somatotrophs and gonadotrophs are easily recognized because they retain the main features described in the pituitary tissue in situ. Other undefined groups of cells bearing small or medium round secretory granules can be identified by immunocytochemistry as PRL, GH or TSH producing cells. The latter technique was critical for the characterization of the hormonal content of secretory granules, the shape, size, electron density and cytoplasmic distribution of which differ substantially from those described in the intact gland. Cells displaying rare small oval or sharp pointed secretory granules were identified as gonadotrophs with anti-LH beta, while corticotrophs showed granules with irregular profiles not previously reported in the gland. These remarkable morphological changes appear to be related to the interruption of the flow of hypothalamic hormones and the disruption of structural and paracrine interrelationships. This investigation reveals that immunocytochemistry is essential for the specific recognition of the various pituitary cell types, and particularly of atypical cells exhibiting morphological features not found in the pituitary gland in situ. PMID- 9341954 TI - Anatomical and surgical aspects of splenic segmentectomies. AB - Based upon the anatomicosurgical segments of the spleen, suggested by DiDio and demonstrated in cadavers, classified and named by Neder (1958) and Zappala (1958, 1959, 1963), the normal segmental organization was anatomically and radiologically confirmed in 51 human spleens, after studying corrosion casts and radiograms of intraparenchymal vessels (Christo, 1959 a, b, 1960, 1962, 1963, 1993). From 1958 to 1965, pioneer segmental resections were performed successfully in 34 dogs and in 9 patients to safely remove traumatic injured splenic segments. At the same time, the overwhelming postsplenectomy infection (OPSI) became well identified. Consequently, to save normally functioning splenic parenchyma became the most important issue in the management of splenic injuries. The anatomical basis for partial splenectomy and splenic segmentectomy is discussed. The term "splenorrhaphy" was employed to designate all conservative or parenchyma saving operations of spleen based upon its vascular supply: from topical packings to splenic sutures including "cappings" and partial splenectomies. From analysis of 38 consecutive reports in 20 years, covering 4,076 patients, it was concluded that "splenorrhaphies" had been electively employed in 46% of the injuries and partial splenectomies were identified in 8.6% of these surgical interventions. However, the critical minimal mass of splenic tissue to be preserved after partial splenectomies is still to be defined. Postoperative complications directly related to "splenorrhaphies" are rare. Uncommonly performed after splenectomies, the heterotopical splenic autotransplantation has presented dubious results. Trials with nonoperative management of splenic blunt trauma injuries have been safer among children, whose spleens are predominantly transversally disrupted and have a higher relationship "capsular resistance/parenchymal bulk". Splenectomies have been most frequently the ultimate result of delayed laparotomy and underlying risks of growing blood requirements may surpass the advantages of preventing OPSI. PMID- 9341955 TI - Morphology of the swine ileum terminale. AB - The morphology of the terminal ileum was studied in 33 male adult pigs of unknown breed. The ileum ended in the cecocolic junction at an acute angle to the cecum. The terminal ileum displayed a cylindrical form (63.6% +/- 8.4 of the cases) more frequently than an ampullary (21.2% +/- 7.1) and/or an infundibular form (15.2% +/- 6.2). The ampullary form is not related to the quadrupedal position of the animal, but is related to the functional phase of the intestine at the time of death. In the mucous tunica of the terminal ileum, longitudinal folds and aggregated lymphoid nodules appeared concentrated along the antimesenteric border. PMID- 9341956 TI - Modified neck muscular system of the giraffe (Giraffa camelopardalis). AB - The muscular and skeletal systems of the long neck were morphologically examined in order to clarify their modification and their functional significance in the giraffe (Giraffa camelopardalis). The longissimus, the thoracic and cervical, spinalis and semispinalis, the cranial and caudal head oblique, and the multifidus muscles, and the nuchal ligament were observed at their origin and insertion. The atlas, axis, and the third cervical vertebra were measured and examined. The modified spinous processes provided the large attachment surface for the strong nuchal ligament and for the muscles of the axis and other cervical vertebrae, while the muscle tendons had their origin in the ventrocaudally enlarged transverse process. It is concluded that the modified muscles with their expanded belly and tendon have the functions of occupying the interspace among long vertebrae, and also of supporting the head and neck by means of their wide attachment to the altered vertebral processes. PMID- 9341957 TI - A new distinctive variation of renal arterial vascularization. AB - In addition to the usual renal arteries, a bilateral artery branching from the aorta was found connecting the aorta to both kidneys in an 82 year-old Caucasian man. By creating an additional blood supply to the kidneys this artery may have had an effect on renal perfusion. PMID- 9341958 TI - The advancement of high-dose chemotherapy and dose intensification schedules. AB - Breast cancer is the most common female cancer--one woman in 12 will have breast cancer at some stage during her life. Early-stage breast cancer is often curable; however, the prognosis is much worse in patients with multiple lymph node involvement or metastatic disease. The overall survival at five years is approximately 60% in women with positive lymph nodes, decreasing to 27%-44% when more than 10 lymph nodes are involved. After metastatic relapse, the mainstay of treatment is palliative. However, recent advances in supportive care have facilitated investigation into the use of dose-intensive chemotherapy regimens. The advancement of high-dose chemotherapy in breast cancer and results from clinical trials in both metastatic disease and the adjuvant setting are reviewed here. The true benefit of high-dose chemotherapy in breast cancer continues to be investigated. It is hoped that the results of worldwide, randomised clinical trials, due within the next three to five years, will provide a clearer indication of the value of high-dose chemotherapy, its costs and the patients whom it will benefit most. PMID- 9341959 TI - Haematological toxicities associated with dose-intensive chemotherapy, the role for and use of recombinant growth factors. AB - High-dose chemotherapy is increasingly accepted as a treatment approach in a number of tumour types. However, there are controversies surrounding its efficacy and there is a need to consider its safety. In view of this, much effort has been directed towards the provision of adequate supportive care strategies to prevent toxicities and to ameliorate myelosuppression. Severe anaemia and its associated symptoms, for example, fatigue can have a debilitating effect on a patient's quality of life and often necessitates red blood cell transfusions. Erythropoietin, a glycoprotein hormone which stimulates red blood cell production, has been established for the treatment of anaemia in patients with chronic renal insufficiency. It is currently approved in most countries for treating anaemia associated with cancer, and its role is emerging especially in patients undergoing high-dose chemotherapy. This paper gives an overview of the studies conducted to date with epoetin alfa (recombinant human erythropoietin) in patients receiving allogeneic and autologous bone marrow transplants or peripheral blood stem cells in conjunction with high-dose chemotherapy. In addition, there are some novel clinical applications for epoetin alfa: for example, in delayed anaemia, as a supportive strategy prior to high-dose chemotherapy and as a synergistic enhancer of blood progenitor cell mobilisation in combination with granulocyte-colony-stimulating factor (G-CSF). PMID- 9341960 TI - Case history. 1: the use of epoetin alfa in delayed anaemia. PMID- 9341961 TI - Case history. 2: The use of epoetin alfa before high-dose chemotherapy. PMID- 9341963 TI - Current strategies for pain control. AB - Pain is the most feared symptom for patients diagnosed with cancer. Although our understanding of cancer pain and its management has greatly improved in the past decade, an unacceptably large proportion of patients still do not receive adequate pain relief. Before commencing any form of treatment, patients must receive a thorough assessment in order to define the pain, causes and severity. The recommendations for progressing a patient from step 2 to step 3 of the WHO analgesic ladder are discussed here as well as the choice of strong opioid substitution. An overview of the benefits of considering alternative routes of administering strong opioids, such as the transdermal delivery of fentanyl (TTS fentanyl), and the use of opioid substitution in patients intolerant to the adverse effects of morphine are also included. Finally, newer approaches to relieving refractory pain, such as neuropathic and bone pain, are considered. PMID- 9341962 TI - The therapy of cancer pain and its integration into a comprehensive supportive care strategy. AB - By the beginning of the 21st century cancer will claim over nine million lives per year. Of these patients, 80%-90% will experience pain at some time during the course of their disease. The management of cancer and the associated pain syndromes is likely to cause a massive drain on resources, making it imperative that we use the resources available to maximum efficiency. In the past decade progress has been made in the understanding of cancer pain and how best to manage it. However, this area of cancer therapy continues to be surrounded by much controversy, particularly the availability and use of strong opioids. Additional measures are required to ensure that no patients are left to suffer the burden of cancer pain. These include: strongly improved education of all healthcare professionals in cancer pain and cancer pain therapy; academic affiliation- university chairs in palliative/supportive care securing education of the coming generations of medical doctors and nurses; improved legislation concerning opioids in many societies through systematic influence of politicians and healthcare decision makers; re-allocation of resources for cancer treatment programmes from the curative to the palliative/supportive treatment spectrum; re defining myths concerning cancer, pain and opioid consumption, both among healthcare workers and in societies in general. The media will play a crucial role in this process. Revising the way in which our resources are allocated and providing all patients access to individually tailored treatment care packages are strategies that could be adopted for future supportive care of cancer patients. PMID- 9341964 TI - Case history: a step up the analgesic ladder. PMID- 9341965 TI - A new method for self-regulation of slow cortical potentials in a timed paradigm. AB - A new method of slow cortical potential (SCP) biofeedback is described, in which subjects were presented with a sequence of two alternating tones. Subjects learned to adjust their SCPs with the 4-s rhythm of presented tones by producing directed SCP changes only in certain inter-tone intervals. Specifically, they learned to simultaneously produce two EEG signals: 1) positive or negative SCP shift at vertex, and 2) SCP asymmetry between the right and the left central area. After one training session, 13 healthy participants were able to differentiate significantly between the negativity and the positivity conditions; this differentiation was achieved within less than 300 ms after the discriminative signal, i.e. much faster than in previous studies employing traditional SCP biofeedback technique. However, these participants did not produce a significant hemispheric asymmetry in the first session. In the second experiment, five subjects participated in prolonged training (6 to 17 sessions). Highly significant control of SCP asymmetry over the precentral cortex was attained in four out of five participants. Advantages and disadvantages of the new method as compared with the "classical" SCP biofeedback technique are discussed. PMID- 9341967 TI - Neocortical dynamics: implications for understanding the role of neurofeedback and related techniques for the enhancement of attention. AB - For nearly 25 years, EEG biofeedback (neurofeedback) has been utilized in research and clinical settings for the treatment and investigation of a number of disorders ranging from attention deficit hyperactivity disorder to seizure disorders as well as many other established and investigational applications. Until recently, mechanisms underlying the generation and origins of EEG have been poorly understood but now are beginning to become much more clarified. Now it is important to combine the information gathered on the genesis of EEG and neocortical dynamics with the findings from neurofeedback investigations. This will help us to develop models of how neurofeedback might operate in producing the changes in EEG and in clinical symptomatology. We know that the cortex operates in terms of resonant loops between neocortical columns of cells known as local, regional, and global resonances. These resonances determine the specific EEG frequencies and are often activated by groups of cells in the thalamus known as pacemakers. There are complex excitatory and inhibitory interactions within the cortex and between the cortex and the thalamus that allow these loops to operate and provide the basis for learning. Neurofeedback is a technique for modifying these resonant loops, and hence, modifying the neurophysiological and neurological basis for learning and for the management of a number of neurologically based disorders. This paper provides an introduction to understanding EEG and neocortical dynamics and how these concepts can be used to explain the results of neurofeedback training and other interventions particularly in the context of understanding attentive mechanisms and for the management of attention deficit/hyperactivity disorders. PMID- 9341966 TI - Respiratory sinus arrhythmia versus neck/trapezius EMG and incentive inspirometry biofeedback for asthma: a pilot study. AB - This pilot study compared biofeedback to increase respiratory sinus arrhythmia (RSA) with EMG and incentive inspirometry biofeedback in asthmatic adults. A three-group design (Waiting List Control n = 5, RSA biofeedback n = 6, and EMG biofeedback n = 6) was used. Six sessions of training were given in each of the biofeedback groups. In each of three testing sessions, five min. of respiratory resistance and EKG were obtained before and after a 20-min biofeedback session. Additional five-min epochs of data were collected at the beginning and end of the biofeedback period (or, in the control group, self-relaxation). Decreases in respiratory impedance occurred only in the RSA biofeedback group. Traub-Hering Mayer (THM) waves (.03-.12 Hz) in heart period increased significantly in amplitude during RSA biofeedback. Subjects did not report significantly more relaxation during EMG or RSA biofeedback than during the control condition. However, decreases in pulmonary impedance, across groups, were associated with increases in relaxation. The results are consistent with Vaschillo's theory that RSA biofeedback exercises homeostatic autonomic reflex mechanisms through increasing the amplitude of cardiac oscillations. However, deep breathing during RSA biofeedback is a possible alternate explanation. PMID- 9341968 TI - Association of apolipoprotein allele epsilon 2 with psoriasis vulgaris in Japanese population. AB - We studied phenotypic variations of apolipoprotein E (apoE) and the corresponding allele frequencies in 100 Japanese patients with psoriasis vulgaris (PV). The phenotypes of apoE were examined using analytical isoelectric focusing followed by immunoblotting with goat anti-apoE antibody and alkaline phosphatase conjugated rabbit antigoat IgG. The phenotypic frequency of apoE3/2 in PV was significantly higher than in healthy controls, and this elevation was associated with an increased frequency of the epsilon 2 allele. Therefore, it is suggested that the apoE molecule plays an important role in the development of PV. PMID- 9341969 TI - Urocanic acid isomers: relation to body site, pigmentation, stratum corneum thickness and photosensitivity. AB - Urocanic acid (UCA), present in the stratum corneum as trans-UCA, absorbs ultraviolet (UV) radiation and isomerizes to cis-UCA. Cis-UCA has been demonstrated to initiate suppression of selected immune responses in several experimental systems. Topical application of UCA-containing products reduces UV induced erythema, but a role for endogenous UCA in photoprotection has not been reported. In this study the relationship between UCA isomers, pigmentation, minimal erythema dose (MED), and stratum corneum thickness was investigated. Pigmentation, concentration of total UCA, and the percentage present as the cis isomer was measured in 36 healthy subjects, skin type I-IV, at six UV-exposed and nonexposed body sites: forehead, chest, back, outer upper arm, inner upper arm, and buttock. The MED was determined by phototesting on buttock skin, and a punch biopsy for measurement of stratum corneum thickness was taken adjacent to the site of the phototest. The percentage of cis-UCA was significantly higher in UV exposed than on nonexposed areas. A small intraindividual variation in total UCA was found, being high on the buttock and the arm, lowest on the forehead. The subject to subject variation of total UCA was considerable at all body sites. No correlation was found between total UCA and MED, skin type, pigmentation, or stratum corneum thickness. PMID- 9341971 TI - Tape stripping of human stratum corneum yields cell layers that originate from various depths because of furrows in the skin. AB - Tape stripping of human stratum corneum is widely used as a method for studying the kinetics and penetration depth of drugs. Several factors can influence the quantity of stratum corneum that is removed by a piece of tape, such as the manner of tape stripping, the hydration of the skin, cohesion between cells, body site and interindividual differences. However, few data are available about the influence of furrows in the human epidermis on the tape-stripping technique. In this study, we investigated the efficacy of tape stripping in removing complete cell layers from the superficial part of the human stratum corneum. A histological section of skin that was tape-stripped 20 times clearly showed nonstripped skin in the furrows, indicating persistent incomplete tape stripping. Replicas of tape-stripped skin surface demonstrated that even after removing 40 tape strips the furrows were still present. We validated the tape-stripping method further with X-ray microanalysis in the mapping mode by scanning electron microscopy, using a TiO2-containing compound as a marker. TiO2 applied to the skin before the tape-stripping procedures was still present after the tenth tape strip, and was specifically located on the rims of the furrows. We emphasize that results from studies using the tape-stripping method have to be viewed from the perspective that cells on one tape strip of the stratum corneum may be derived from different layers, depending on the position of the tape strip in relation to the slope of the furrow, and such results should be interpreted with considerable caution. PMID- 9341970 TI - Stratum corneum swelling. Biophysical and computer assisted quantitative assessments. AB - The aim of this study was to characterize the swelling behaviour of the stratum corneum. Stratum corneum pieces isolated from the breast region of 20 different females were incubated in distilled water at two different temperatures (20 degrees C and 45 degrees C) for 90 min and 24 h, respectively. Half of the stratum corneum pieces were previously extracted with chloroform-methanol (2:1). The area-enlargement was photographically recorded. The thickness enlargement was determined using a confocal laser scanning microscope. The average swelling (99% confidence interval) in the area dimension at 20 degrees C was 8.4% +/- 1.4% (n = 20), which corresponded to an average swelling in the length (lateral) dimension of approximately 4.1%. The swelling in the thickness dimension was 26.3% +/- 16.3% (n = 8). The swelling was most pronounced in the thickness dimension and was complete after 90 min of water immersion (P < 0.01, n = 5). In addition, the removal of the intercellular lipids with chloroform/methanol (2:1) induced a decreased swelling in the samples (P < 0.01, n = 20). An increase in temperature of the water from 20 degrees C to 45 degrees C resulted in an increase in swelling (P < 0.01, n = 20). Taken together our results support the idea that the mechanism of stratum corneum swelling is linked to the intercellular lipid structure and hence to skin barrier function. PMID- 9341973 TI - The role of granzyme B-expressing CD8-positive T cells in apoptosis of keratinocytes in lichen planus. AB - Epidermal basal cell injury with colloid body formation is a characteristic feature of lichen planus. Infiltrated cells are thought to be responsible for the epidermal injury. Ultrastructural findings of colloid bodies are typical of apoptosis. Granzymes in cytotoxic T lymphocytes are involved in apoptosis probably together with perforin. Based on this background, we analyzed the role of granzyme B in the mechanisms of epidermal injury in lichen planus. On electron microscopy, basal and suprabasal cells showed condensed chromatin and fragmented nuclei which are typical morphological features of apoptosis. Nuclei of colloid bodies were positively stained by the in situ nick end labeling technique indicating that colloid bodies are subsequently formed in the process of apoptosis. Immunohistochemical staining showed CD8-positive infiltrating cells to contain granzyme B. Cells undergoing exocytosis also contained granzyme B. By immunoelectron microscopy, granzyme B molecules were observed to be secreted from a lymphocyte to an apoptotic keratinocyte. These findings suggest that granzyme B positive CD8 cells seem to induce apoptosis of keratinocytes in lichen planus. PMID- 9341972 TI - Keratin and involucrin expression in discoid lupus erythematosus and lichen planus. AB - In the present study, keratin and involucrin expression were studied in cutaneous lesions of discoid lupus erythematosus and lichen planus in order to gain a better understanding of the abnormal differentiation or maturation of the epidermal cells in these dermatoses. Ten specimens each from discoid lupus erythematosus and lichen planus were analyzed by immunohistochemical techniques, using a panel of monoclonal antikeratin antibodies and polyclonal anti-involucrin antibody, and five specimens each were analyzed by one- and two-dimensional gel electrophoresis and immunoblot analysis using three antikeratin antibodies. No significant difference was found between the dermatoses. The expression of differentiation-specific keratins showed a similar pattern to that in normal epidermis, and involucrin was expressed even in the lower part of the stratum spinosum. Keratins 6 and 16, which are characteristic markers of hyperproliferative states, and keratin 17 were detected in nonhyperproliferative and atrophic epidermis with hydropic degeneration and inflammatory infiltrates in the dermis. These results suggest that expression of keratins 6, 16 and 17 in discoid lupus erythematosus and lichen planus may reflect a wound healing response to the damage to the basal cell layer, or may be under the control of cytokines produced by infiltrating inflammatory cells in the dermis. PMID- 9341974 TI - Retinoic acid attenuates phospholipase C-mediated signaling in HaCaT keratinocytes. AB - Release of inositol(1,4,5)trisphosphate (Ins(1,4,5)P3) generated by phospholipase C (PLC) upon receptor stimulation plays an important role in the regulation of cell growth and differentiation. A second, completely different, signal transduction system involves retinoic acid (RA) and related derivatives. Binding to intracellular receptor sites can modulate keratinocyte growth and inhibits differentiation. The present study was aimed at characterizing possible interactions between the two signalling pathways in HaCaT keratinocytes. As determined by anion exchange chromatography and HPLC analysis, HaCaT keratinocytes treated with 1 microM RA for up to 72 h showed a marked decrease in Ins(1,4,5)P3 release upon stimulation with 10 microM bradykinin or 10 microM ionomycin. Thin-layer chromatography of phosphatidylinositol phosphates, the substrates of PLC, revealed no differences between RA-treated and untreated cells. Western blot analysis of the PLC isozymes present in HaCaT cells, PLC beta 3 and PLC gamma 1, showed no alterations in the expression of these proteins in RA-treated cells as compared to vehicle-treated controls. In addition, expression of the PLC-activating G protein G alpha q was not affected by RA treatment. Our results show that RA downregulates the PLC-mediated signaling system. The point of interference of this signal transduction crosstalk has yet to be elucidated. Our results suggest, furthermore, that RA-induced attenuation of keratinocyte differentiation might be mediated at least in part by the downregulation of Ins(1,4,5)P3 release. PMID- 9341975 TI - SCH 47112, a novel staurosporine derivative, inhibits 12-O-tetradecanoylphorbol 13-acetate-induced inflammation and epidermal hyperplasia in hairless mouse skin. AB - Protein kinase C (PKC) regulates keratinocyte growth and differentiation as well as inflammation in skin, processes which are abnormal in skin diseases such as psoriasis. 12-O-tetradecanoylphorbol-13-acetate (TPA) binds to and activates PKC. We investigated the effects of SCH 47112, a novel staurosporine derivative, which interacts with the catalytic domain of PKC, on TPA-induced inflammation and hyperplasia in hairless mouse skin and TPA-induced differentiation in cultured human keratinocytes. Dorsal mouse skin was treated with vehicle, TPA (2.0/ 2.5 nmol) or SCH 47112 followed by TPA. Epidermal thickness, and epidermal, upper dermal and deep dermal inflammation (assessed on an ordinal semiquantitative scale) were determined in biopsies taken 24 h and 48 h post-treatment. SCH 47112 (100 nmol) inhibited TPA-induced epidermal, upper dermal and deep dermal inflammation by 71%, 45% and 22%, respectively, at 24 h (n = 3, P < 0.05). TPA induced epidermal hyperplasia was inhibited by SCH 47112 (400 nmol) by 38% at 48 h (n = 3, P < 0.05). In addition, in cultured human keratinocytes, SCH 47112 inhibited TPA induction of transglutaminase. I protein, which catalyzes the formation of crosslinked envelopes. These results indicate that SCH 47112 exhibits biological activity, inhibiting TPA-induced changes in hairless mouse skin in vivo and cultured human keratinocytes in vitro, and suggest that PKC inhibitors may have a therapeutic role in inflammatory skin diseases. PMID- 9341976 TI - Decreased responsiveness of T cells to toxic shock syndrome toxin-1 in patients with severe psoriasis at active stage. PMID- 9341977 TI - Psoriatic fibroblasts enhance cornified envelope formation in normal keratinocytes in vitro. PMID- 9341978 TI - Hair cycle-dependent changes in the gene expression and protein content of transforming factor beta 1 and beta 3 in murine skin. PMID- 9341979 TI - Molecular unraveling of von Willebrand's disease: still some way to go. PMID- 9341980 TI - Epinephrine and platelet function. PMID- 9341981 TI - Iron chelation by gene therapy: a twenty-first century challenge for molecular medicine. PMID- 9341982 TI - The need for improved diagnosis of heparin-induced thrombocytopenia. PMID- 9341983 TI - Potassium channels in hypertension: homeostatic pathways to buffer arterial contraction. PMID- 9341984 TI - von Willebrand disease and quantitative variation in von Willebrand factor. PMID- 9341985 TI - Effect of low concentration of epinephrine on human platelet aggregation analyzed by particle counting method and confocal microscopy. AB - The effect of a physiologic concentration of epinephrine (Ep) on platelet activation was studied by using a novel method that simultaneously measures platelet aggregation by changes in light transmission and counts particles of various sizes by using light scattering. Detailed morphologic changes associated with activation process were studied by using confocal laser scanning microscopy (CLSM). A low concentration of Ep (20 nmol/L) corresponding to a high physiologic concentration triggered the formation of small platelet aggregates (diameter 7 to 30 microm) without any change in light transmission. The redistribution of filamentous actin (F-actin) and the expression of activated glycoprotein IIb-IIIa complex (GPIIb-IIIa), detected by PAC-1 binding, were also observed in the platelets comprising the small aggregates. Attempts were then made to detect changes in cytoplasmic ionized Ca2+ ((Ca2+)i) in individual platelets involved in the aggregate formation by CLSM with fluo-3, a Ca2+-indicating dye. Ep caused a weak (Ca2+)i increase in some individual platelets involved in the formation of small aggregates. This (Ca2+)i increase was associated with platelet aggregation, because no (Ca2+)i rise was detected in single platelets. Furthermore, platelets stimulated by Ep in the presence of RGDS or Ro 44-9883, a GPIIb-IIIa antagonist, did not form small aggregates or trigger a (Ca2+)i rise. Prior incubation with low concentrations of Ep (20, 100 nmol/L) enhanced the initial formation of small (diameter 7 to 30 microm), medium (diameter 30 to 50 microm), or large (diameter 50 to 70 microm) aggregates induced by a subthreshold concentration of adenosine diphosphate (0.25 micromol/L) as determined by the particle counting method. However, no apparent synergistic (Ca2+)i increase was observed in platelets involved in aggregate formation. From these observations the following conclusions have been reached. (1) A high physiologic concentration of Ep (20 nmol/L) is capable of triggering the formation of small aggregates, resulting in the redistribution of F-actin and the expression of activated GPIIb-IIIa complex. (2) An increase in (Ca2+)i is observed in platelets comprising the small aggregates. This increase is not related to the binding of Ep to its receptor but most likely is triggered by platelet-platelet association. (3) The characteristic potentiating effect of Ep is not due to the synergistic increase in (Ca2+)i. PMID- 9341986 TI - Alteration in the organ distribution of iron by truncated transferrin: implications for iron chelation therapy. AB - The ability of the partial molecule of transferrin, truncated transferrin (t-Tf), to act as an excretable biologic iron chelator was examined. We confirmed the observations of Zak and Aisen (Zak O, Aisen P. Biochem Biophys Acta 1985;1952:24 8) that thermolysin treatment of human transferrin produces half molecules that retain iron-binding capacity. These molecules are poorly recognized by surface receptors on either human or murine cells. Although the plasma half-life of human transferrin in mice is moderately long (40 hours), injection of t-Tf into mice results in its rapid clearance (half-life = 10 minutes). Injection of iron 59 labeled transferrin results in the deposition of iron in the major hematopoetic organs of mice such as the spleen, bone marrow, and liver. Injection of 59Fe labeled t-Tf results in the quantitative recovery of iron in the kidneys: 59Fe is retained in the kidney for substantial periods of time with little evidence of its excretion into urine. Injection of iodine 125-labeled t-Tf also results in the deposition of radioactivity in the kidneys, but 125I is rapidly excreted into the urine, where it is detected as free iodine. These results indicate that although t-Tf is directed to the kidney and filtered by the glomerulus, the molecule is reabsorbed and degraded, and iron is retained. These results have implications in the design of iron chelators. PMID- 9341989 TI - Serum vitamin E decreases in HIV-seropositive subjects over time. AB - Vitamin E is an important lipid soluble antioxidant that has a number of crucial functions including protecting lipids from oxidative damage. It also may play an important role in enhancing the immune response in subjects with Human Immunodeficiency Virus (HIV) infection. The current study measured the serum level of vitamin E in 121 HIV seropositive subjects with no prior pulmonary complications. Although the mean level was normal at 9.0 +/- 0.5 microg/ml, 22.3% of the subjects had a deficient level of less than 5 microg/ml. In addition, 42 subjects were studied longitudinally and serum vitamin E levels were determined at baseline and 12 months later. The mean serum vitamin E level in this group significantly decreased after 12 months compared with baseline levels (5.9 +/- 0.5 microg/ml compared with 9.6 +/- 0.9 microg/ml, p = 0.001). The CD4 counts also were significantly decreased after 12 months (460.6 +/- 36.0 cells/mm3 versus 390.5 +/- 37.7 cells/mm3, p = 0.032). No significant correlations were observed between the decrease in serum vitamin E and the change in CD4 count, body mass index (BMI), or serum albumin levels over the 12-month period. In conclusion, a significant portion of HIV-seropositive subjects have a deficiency in serum vitamin E early in the course of their disease. Furthermore, there is a significant decrease in serum vitamin E levels in these subjects over 12 months. PMID- 9341988 TI - Defective elimination of C3b/iC3b-coated autologous erythrocytes in patients with primary biliary cirrhosis, alcoholic cirrhosis, and ulcerative colitis. AB - Delayed in vivo elimination of autologous erythrocytes coated with immunoglobulin G has been reported in autoimmune and inflammatory gastrointestinal diseases. Our aim was to elucidate whether impairment of the macrophages was restricted to the Fc receptors of the reticuloendothelial system or whether complement receptors were also affected. We studied elimination by complement receptors of autologous erythrocytes coated with fragments of C3 and C4 in patients with primary biliary cirrhosis, ulcerative colitis, and alcoholic cirrhosis. Impaired function was seen in all patient groups as compared with function in normal subjects, both concerning the mean half-life of the injected cells and the total number of eliminated erythrocytes. Neither of these parameters correlated with the levels of C3 fragments bound to the injected autologous erythrocytes. This is the first report of defective complement receptor function in ulcerative colitis and alcoholic cirrhosis. Immunoglobulin G-dependent elimination of erythrocytes was confirmed to be lowered in all patient groups. The results suggest severe macrophage functional aberrations involving both complement receptors and Fc receptors as the basis of phagocytic defects in autoimmune/inflammatory conditions. In contrast, a general loss of macrophages might cause the functional loss in alcoholic cirrhosis. PMID- 9341987 TI - Activation of platelets by sera containing IgG1 heparin-dependent antibodies: an explanation for the predominance of the Fc gammaRIIa "low responder" (his131) gene in patients with heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a prothrombotic disorder caused by heparin-dependent IgG (HIT-IgG) that recognizes a complex of heparin and platelet factor 4 (PF4), leading to platelet activation via the platelet Fc gammaIIa receptors (Fc gammaRIIa). Not all patients who generate HIT-IgG in response to heparin develop HIT, however, possibly because of observed differences in the ability of platelets from healthy individuals to be activated by HIT sera. It is known that a polymorphism in the platelet Fc gammaRIIa plays an important role in determining platelet reactivity to murine platelet-activating monoclonal antibodies of the IgG1 subclass: homozygous arg131 ("high responder" or HR) platelets respond well, and homozygous his131 ("low responder" or LR) platelets respond poorly, respectively, to these murine monoclonal antibodies. We sought to determine whether the differing risk for HIT among patients who receive heparin, as well as the variable platelet reactivity to HIT sera, could be explained by preferential activation by HIT-IgG of platelets bearing a particular Fc gammaRIIa phenotype. We found that the LR Fc gammaRIIa gene frequency was significantly overrepresented among 84 HIT patients, compared with that of 264 control subjects (0.565 versus 0.471; p = 0.03). We studied the subclass distribution of HIT-IgG against its major antigen, heparin/PF4 complexes, and found that 55 of 61 (90%) HIT sera expressed IgG1 antibodies either alone (n = 47) or in combination with IgG2 (n = 5) or IgG3 (n = 3). We then compared the platelet-activating profile of HIT sera with murine platelet-activating monoclonal antibodies. As expected, the murine IgG1 monoclonal antibodies preferentially activated platelets from homozygous HR individuals. In contrast, however, the LR homozygous platelets exhibited the greatest reactivity to HIT sera that contained predominantly anti heparin/PF4 antibodies of the IgG1 subclass. We conclude that the significant overrepresentation of the LR (his131) gene among patients with HIT may be explained by the preferential activation of LR Fc gammaRIIa platelets by HIT antibodies of the IgG1 subclass, which is the predominant immunoglobulin subclass generated in HIT. PMID- 9341990 TI - Standardization of a single-cell assay for sensitive detection of multidrug resistance protein expression in normal and malignant cells in archival clinical samples. AB - Multidrug resistance protein (MRP), like P170, confers multidrug resistance, but its clinical relevance is uncertain, whereas P170 is an accepted cause of chemotherapy failure for which ongoing reversal trials are being conducted. Because such trials have been only modestly successful, we must investigate alternative drug resistance mechanisms such as MRP, which is poorly blocked by P170 inhibitors. The significance of MRP has remained undefined because MRP mRNA is difficult to assay in archival material, does not necessarily reflect MRP levels, and is widely expressed in normal or hematopoietic cells within tumors and bone marrow. Because conventional immunoblot or immunocytochemistry may not be sensitive enough to detect low or heterogeneous MRP expression in clinical samples, we elected to score MRP in single tumor cells by modifying our P170 assays that have proven valuable for correlating P170 expression with the outcome of pediatric cancer chemotherapy. We enhanced the signal-to-noise ratio with several peroxidase-tagged secondary antibody layers and staining refinements, standardizing the assay with MRP-negative and MRP-positive but P170-negative transfected or drug-selected controls in which MRP was quantified by immunoblot. We confirmed sensitivity by staining a very low MRP-expressing revertant line and "mixed" samples containing small numbers of positive cells; we confirmed specificity by applying two antibodies directed against separate MRP epitopes. We examined neuroblastoma, osteosarcoma, rhabdomyosarcoma, and retinoblastoma samples, identifying MRP-positive malignant cells, which were distinguishable from MRP-positive normal cells. This assay may be valuable for early diagnosis of low but potentially important MRP expression, which would allow timely application of alternative therapy, perhaps with MRP-specific blockers. PMID- 9341991 TI - Role of granulocyte elastase in indomethacin-induced gastric mucosal lesion formation in rats. AB - To investigate whether granulocyte elastase may be involved in indomethacin induced gastric mucosal injury, we examined the effects of the granulocyte elastase inhibitors ONO-5046 and L-658,758 on gastric mucosal lesion formation in rats given indomethacin. Both gastric mucosal lesion formation and gastric mucosal vascular damage were markedly attenuated in animals with leukocytopenia and in those given granulocyte elastase inhibitors. The administration of indomethacin significantly increased gastric myeloperoxidase activity, a measure of leukocyte accumulation, 3 hours after the administration of indomethacin compared with activity in animals receiving saline solution. The administration of ONO-5046 or L-658,758 significantly prevented this increase. Histologic examinations revealed submucosal edema and marked infiltration by leukocytes, as well as widespread necrosis with loss of surface epithelium. ONO-5046 and L 658,758 markedly prevented these histologic changes. Although cimetidine significantly prevented mucosal lesion formation, it did not inhibit either granulocyte elastase release from activated neutrophils in vitro or gastric accumulation of leukocytes in vivo. These results suggest that granulocyte elastase as well as gastric acid may play an important role in the pathologic process by which indomethacin induces gastric mucosal lesions. PMID- 9341992 TI - Effects of diaspirin-cross-linked hemoglobin (DCLHb) on the microcirculation of striated skin muscle in the hamster: a study on safety and toxicity. AB - Hemoglobin-based oxygen-carrying solutions are reported to exert vasoconstrictor effects and to enhance oxygen radical formation, particularly during ischemia reperfusion. This study investigates whether diaspirin-cross-linked hemoglobin (DCLHb) affects the microvascular integrity of striated skin muscle. The microcirculation model in the hamster and intravital fluorescence microscopy were applied for investigation of the microvascular changes in striated skin muscle. Hypervolemic infusion (500 mg x kg(-1), I.V.) and isovolemic exchange transfusion (3.3 gm x kg(-1) I.V.; hematocrit 30%) with DCLHb (1) led to a short-lasting (0 to 2 minutes) arteriolar constriction (approximately 20% reduction in baseline diameter), (2) significantly influenced arteriolar vasomotion, (3) increased venular red blood cell velocity by 1.5-fold (p < 0.05 vs dextran, Mr 60,000), and (4) did not enhance microvascular leukocyte-endothelium interaction or endothelial permeability. Resuscitation from severe hemorrhagic shock with autologous blood (AuB) or DCLHb (33 ml x kg(-1), I.V.) immediately restored mean arterial pressure and heart rate, whereas 6% dextran (60 kd)(Dx-60) did not return these parameters to baseline. Venular red blood cell velocity was restored to 110% of baseline after DCLHb, to 90% of baseline after AuB, and to 45% of baseline after Dx-60. Leukocyte-endothelium interaction was significantly enhanced after resuscitation with AuB and Dx-60, whereas this phenomenon was absent after DCLHb. These data demonstrate that DCLHb increases venular red blood cell velocity under both nonischemic and postischemic conditions without inducing enhanced leukocyte-endothelium interaction in the microcirculation of striated skin muscle. PMID- 9341993 TI - Effects of diaspirin-cross-linked hemoglobin (DCLHb) on local tissue oxygen tension in striated skin muscle: an efficacy study in the hamster. AB - Using the dorsal skin fold chamber model in the hamster, we analyzed local tissue partial oxygen pressure (PO2) in the striated skin muscle under nonischemic and postischemic conditions with a Clark-type multiwire oxygen surface electrode. Hypervolemic infusion (500 mg x kg(-1) I.V.) or isovolemic exchange transfusion (3.3 gm x kg(-1) I.V.; hematocrit 30%) with diaspirin-cross-linked hemoglobin (DCLHb) resulted in a slight decrease of the mean value of the local tissue PO2 (mm Hg) 1 hour after administration. Concomitantly, the frequency distribution curves of local tissue PO2 values were found to be more narrow (fewer values > 25 mm Hg and < 10 mm Hg). Resuscitation from severe hemorrhagic shock (bleeding of 33 ml x kg(-1) at 0.4 ml x min(-1)) with autologous blood (AuB), Dx-60, or DCLHb led to an increase of mean tissue PO2 values by 4.2-fold (p < 0.05 versus Dx-60), 1.9-fold, and 3.7-fold (p < 0.05 versus Dx-60), respectively, 2 hours after resuscitation. The reduction of tissue hypoxia (0-5 mm Hg) was significant only in the AuB- and DCLHb-treated animals. This study indicates that DCLHb effectively reverses tissue hypoxia after resuscitation from severe hemorrhagic shock by inducing a more homogeneous distribution of the local tissue PO2 levels. PMID- 9341994 TI - Inter-alpha-inhibitor as marker for neutrophil proteinase activity: an in vitro investigation. AB - Human neutrophil proteinases have been implicated in the pathogenesis of a wide variety of inflammatory diseases. The degradation of plasma proteins such as coagulation and fibrinolysis factors has been attributed to the excessive release of elastase in septicemia and in other conditions in which heightened proteolysis occurs. Inter-alpha-inhibitor (IalphaI) is particularly sensitive to cleavage by leukocyte proteinases. For this reason, the determination of IalphaI has been proposed as a method for evaluating plasma protein proteolysis by neutrophil enzymes. In this article we provide evidence that intact residual IalphaI can be accurately quantified by enzyme-linked immunosorbent assay (ELISA) determination without interference from fragments released from IalphaI by incubation with triggered neutrophils. We demonstrate that under these conditions IalphaI was quickly and steadily proteolyzed in a cell dose-dependent manner. Alpha-1 proteinase inhibitor (alpha1PI) partially protected IalphaI; however, the proteolysis persisted when IalphaI was incubated with stimulated neutrophils in the presence of a large relative excess of alpha1PI over the amount of elastase theoretically present in cells. For the same amount of alpha1PI, serum provided a better protection than alpha1PI alone but did not completely inhibit the IalphaI degradation. Therefore, ELISA determination of IalphaI might be useful for monitoring the in vivo activity of neutrophil proteinases in systemic proteolytic states. PMID- 9341995 TI - Single channel openings in a vascular smooth muscle cell. PMID- 9341996 TI - In search of the ideal measure of high-density lipoprotein. PMID- 9341997 TI - Another step toward standardization of methods for measuring hemoglobin A1c. PMID- 9341998 TI - Molecular diagnosis of B- and T-cell lymphomas: fundamental principles and clinical applications. AB - Molecular diagnostic assays have become routine in the evaluation of lymphoid malignancies. Both Southern transfer and polymerase chain reaction (PCR) technologies are used to assess for B- and T-cell clonality, the presence of rearrangements involving protooncogenes such as bcl-1 and bcl-2, and the monitoring of minimal residual disease. We review the fundamentals of B- and T cell ontogeny as well as the basic principles of the Southern transfer and PCR assays and their applications to the diagnosis of lymphoid malignancies. PMID- 9341999 TI - On the meaning of "sensitivity". AB - The term "sensitivity" (as applied to an analytical method's performance) has again become a subject of controversy. Certain authorities (e.g., IUPAC) define a system's sensitivity as the response curve slope (or response/dose), others (e.g., IFCC) in terms of the detection limit. Many investigators have failed to perceive the contradiction between these concepts, wrongly assuming that maximizing "sensitivity" in the first sense maximizes it in the second (i.e., that they are inversely related). The existence of different meanings for this term (when used in the present context) is a source of confusion that has, among other things, led to erroneous ideas relating to immunoassay design. Such confusion should be terminated by adoption of one or the other of the definitions. However, the definitions are not of equal merit. We advance arguments against retention of the "slope" definition, which conflicts with the word's common meaning and is meaningless as an indicator of the performance of a measuring system. PMID- 9342000 TI - The inseparable triad: analytical sensitivity, measurement uncertainty, and quantitative resolution. AB - The formal definition of sensitivity associates the term with the change in the response of a system for a small change of the stimulus causing the response, i.e., the ratio of the response of a system to the stimulus causing it. One interpretation of sensitivity associates the rate of change of the response for a small change of the stimulus as the slope of a calibration plot of response vs stimulus. An alternative interpretation associates sensitivity with the smallest value of the stimulus that can be resolved with a given degree of confidence, i.e., the detection limit. Applications of the first usage to analytical chemistry date at least to the beginning of this century; applications of the second interpretation are of more recent origin. The accompanying paper argues in favor of the second interpretation on the basis that, among other things, the "slope" interpretation conflicts with the formal definition of sensitivity and is meaningless as a descriptor of the performance of a measuring system. In this paper I offer arguments to support my belief that the slope definition of sensitivity is consistent with both formal definitions and accepted usage in analytical chemistry and, more importantly, that it is an invaluable descriptor of one of the most important characteristics of any analytical method. I include information to support my belief that proper use of the slope definition yields much more information than is available in the "detection limit" interpretation. PMID- 9342001 TI - Receiver operating characteristic plots to evaluate Guthrie, Wallac, and Isolab phenylalanine kit performance for newborn phenylketonuria screening. AB - We used ROC plots to evaluate the clinical performance of the Guthrie, Wallac, and Isolab assays for newborn phenylketonuria (PKU) screening and assessed the screening discriminatory power of these three assays by the area under the ROC plot, Youden's J index, and the likelihood ratio. The use of these plots not only allows us to pinpoint the exact cutoff value in screening, but also provides a direct comparison of these three different assays in clinical outcome performance. The optimum cutoff for the newborn PKU screening is a blood phenylalanine concentration of 0.30, 0.27, and 0.18 mmol/L for the Guthrie, Wallac, and Isolab assays, respectively. We conclude that the Wallac and Isolab kits, like the Guthrie assay, are suitable for newborn PKU screening. PMID- 9342002 TI - Quantitative polymerase chain reaction for human herpesvirus diagnosis and measurement of Epstein-Barr virus burden in posttransplant lymphoproliferative disorder. AB - Human herpesviruses can cause acute diseases such as chicken pox or mononucleosis, but also may reactivate during immunosuppression and result in severe or life-threatening illnesses such as shingles or lymphoproliferative disorders. We report the development and validation of a quantitative PCR method to measure viral burden for all eight human herpesviruses (HSV1, HSV2, VZV, EBV, CMV, HHV6, HHV7, and KSHV) in patients' samples. The method uses an internal standard that is coamplified with the viral target, allowing quantification of viral genomes in absolute terms (e.g., viral targets/mL of blood) and ruling out false-negative results. We demonstrate that transplant patients with lymphoproliferative disorder carry an EBV viral burden 3 logs higher than nontransplant patients. EBV titers in transplant patients without a lymphoproliferative disorder are between these values. This quantitative PCR method may aid in differentiating clinically significant vs latent viral burden in immunosuppressed patients. PMID- 9342003 TI - Combinations of beta chain abnormal hemoglobins with each other or with beta thalassemia determinants with known mutations: influence on phenotype. AB - Hematological and hemoglobin (Hb) data are presented for numerous patients with compound heterozygosities for different beta chain variants and for a beta chain variant with different beta-thalassemia (beta-thal) alleles. Considerable variations, which result from the type of beta chain variant and beta-thal mutation, can be noted. The comparison again emphasizes the importance of determining the diagnoses at the molecular level to aid the physician in the management of patients with different combinations of abnormalities. Simplification and commercialization of modern technology may make the introduction of this approach in some clinical chemistry laboratories possible. PMID- 9342004 TI - Direct detection of multiple point mutations in mitochondrial DNA. AB - Mitochondrial defects can be caused by mutations in nuclear or mitochondrial DNA. Large deletion/duplication and point mutations are the two major types of mitochondrial DNA (mtDNA) mutations. Comprehensive molecular diagnosis requires the analysis of multiple point mutations. We developed an effective multiplex PCR/allele-specific oligonucleotide (ASO) method to simultaneously screen multiple point mutations in mtDNA. The system involved three pairs of primers to amplify mutation "hot spots" at tRNA(leu(UUR)), tRNA(lys)/ATPase, and ND4 regions, followed by detection of point mutations with ASO probes. Over 2000 specimens were analyzed and the results were compared with those from previous studies with the PCR/restriction fragment length polymorphism method. Our data demonstrate that the multiplex PCR/ASO method is much more sensitive in the detection of low mutant heteroplasmy. It is simple and cost effective, especially if a large number of samples are to be screened for multiple point mutations. PMID- 9342005 TI - Prostate-specific antigen in serum during the menstrual cycle. AB - We previously found that prostate-specific antigen (PSA) expression in the female breast is regulated by steroid hormones and their receptors. We have now examined whether the PSA concentration in serum changes during the menstrual cycle of healthy women. Among 14 women studied, 3 had serum PSA > or = 4 ng/L; their changes in PSA content during the menstrual cycle were studied in 7 informative cycles. We found that PSA concentrations in serum are highest during the mid- to late follicular phase, drop continuously with a half-life of 3-5 days between the late follicular phase and mid-cycle, and reach a minimum during the mid- to late luteal phase. PSA changes do not correlate with changes in lutropin (LH), follitropin (FSH), or estradiol concentrations. However, PSA peaks seem to follow the progesterone concentration peaks, with a delay of 10-12 days. Sera of some volunteers were tested for their ability to upregulate PSA protein and PSA mRNA in a tissue culture system based on the T-47D breast carcinoma cell line. Only sera obtained during the mid- to late luteal phase were able to upregulate the PSA mRNA and protein. In stimulation experiments in vitro, progesterone, but not LH, FSH, estradiol, human chorionic gonadotropin, prolactin, or growth hormone, was able to upregulate PSA mRNA and protein in the T-47D cell line. These data suggest that PSA is produced in a cyclical manner during the menstrual cycle. PMID- 9342006 TI - ELISA determination of soluble urokinase receptor in blood from healthy donors and cancer patients. AB - Measurement of urokinase receptor (uPAR) in tumor extracts has prognostic value, but assay of the soluble uPAR (suPAR) in peripheral blood may offer wider applications in cancer patient management. A tumor extract uPAR ELISA was modified to eliminate nonspecific plasma protein interference, enabling specific detection of suPAR in plasma and sera with >90% recovery of added calibrator. suPAR concentrations in citrate plasma correlated with sera in 93 healthy blood donors (r = 0.84, P <0.0001), with a median value for both of 1.2 microg/L. The plasma median for 19 advanced breast cancer patients was 2.9 microg/L suPAR, and a similar increase was found for 10 advanced colon cancer patients, consistent with release of suPAR from tumors into blood. Repetitive monitoring of suPAR in cancer patients' blood may have value in assessment of prognosis and tumor recurrence. PMID- 9342007 TI - Multicenter evaluation of a second-generation assay for cardiac troponin T. AB - We report on the evaluation of the second-generation assay for cardiac troponin T (cTnT) on the Enzymun system. This new assay is completely specific for the cardiac isoform of TnT, utilizing two cardiospecific monoclonal antibodies. The assay time is reduced to 45 min. The interassay precision shows a median CV of 5.5%; 20% interassay CV was found between 0.05 and 0.1 microg/L. The cardiosensitivity of the second-generation cTnT assay in patients with ischemic myocardial injury appears equivalent when compared with the first-generation assay. We found no falsely positive results in patients with skeletal muscle damage including multitraumas, surgery patients, and marathon runners who showed highly increased values with the unspecific first-generation assay. In Duchenne disease cTnT was still increased, but to a much lower extent. cTnT remains increased in renal failure, but to a lesser degree than with the first-generation assay. The cause of this increase remains unclear. Although a cross-reactivity of skeletal muscle TnT in the second-generation assay could be excluded by our findings, minor myocardial damage or expression of the cardiac isoform of TnT in regenerating muscles cannot be ruled out in those cases with apparently falsely increased cTnT values. The second-generation cTnT assay is a step forward in the combination of cardiosensitivity and cardiospecificity in biochemical markers for diagnosis of heart disease. PMID- 9342008 TI - Evaluation of precipitation and direct methods for HDL-cholesterol assay by HPLC. AB - HDL-cholesterol (HDL-C) values measured by precipitation (sodium phosphotungstate MgCl2) and direct methods were compared with those obtained by HPLC with a new column (TSKgel Lipopropak) and an eluent (TSKeluent LP-1). The HDL-C values determined by the precipitation method were significantly (P <0.001) lower than those by the HPLC method, whereas the HDL-C values by the direct method were slightly but significantly higher (P <0.02) than those by the HPLC method. A quantitative HPLC analysis of the cholesterol concentration in HDL and non-HDL fractions in the supernatant of serum separated by precipitation reagents with different MgCl2 concentrations ranging from 7.3 to 44 mmol/L revealed that reagents with >22 mmol/L MgCl2 precipitated part of HDL as well as non-HDL lipoproteins. The HPLC method providing quantitative and qualitative information with high precision was regarded as being a reliable approach for HDL-C assay. The HPLC can be also used to evaluate alternative methods for cholesterol assay. PMID- 9342010 TI - Determination of total cholesterol in serum by liquid chromatography-isotope dilution mass spectrometry. AB - We have developed a liquid chromatography-isotope dilution mass spectrometry procedure to quantify total cholesterol in serum. A particle-beam interface was used for coupling the liquid chromatograph and the mass spectrometer. After electron impact ionization the ions m/z = 386 and m/z = 389 were used for selective ion monitoring of cholesterol and the internal standard [25,26,27 (13)C]cholesterol. The sample preparation steps required for serum materials are alkaline hydrolysis and an extraction of the cholesterol into the cyclohexane phase. Imprecision for the determination of cholesterol in control materials is typically <1.0%. The deviation from the certified reference values was <0.75% for all control materials tested. A method comparison of the results obtained by this method with those obtained by gas chromatography-isotope dilution mass spectrometry for n = 28 pooled human sera derived from samples analyzed in our routine laboratory did not show differences >2.5%. PMID- 9342009 TI - Relation between lipoprotein(a) concentrations in patients with acute-phase response and risk analysis for coronary heart disease. AB - This study investigated whether lipoprotein(a) [Lp(a)] is an acute-phase reactant that can cause important bias in risk factor analysis for coronary heart disease among patients with an acute-phase response (APR patients). To determine whether serum Lp(a) concentrations increase among APR patients, we compared the Lp(a) concentrations and apolipoprotein(a) [apo(a)] phenotypes of 100 controls with those of a random sampling of 100 APR patients. Serum Lp(a) concentration was measured by ELISA; Lp(a) phenotyping was performed by electrophoresis on sodium dodecyl sulfate-polyacrylamide gel. Lp(a) was significantly (P <0.0001) higher among APR patients (mean +/- SD 0.300 +/- 0.284 g/L) than among controls (0.118 +/- 0.193 g/L) even though the distribution of apo(a) phenotypes did not differ significantly. The 100 APR patients were grouped into 4 categories: 48 patients with infections, 25 postoperative patients, 17 patients with tumors, and 10 patients with other diseases, all of whom showed substantially higher Lp(a) values than did the controls. For the S5, S4S5, S5S5, and S4 phenotypes, the mean concentrations of serum Lp(a) were substantially higher among the APR patients. PMID- 9342011 TI - Measurement of low-density lipoprotein particle size by high-performance gel filtration chromatography. AB - We describe a new technique for measuring LDL size by high-performance gel filtration chromatography (HPGC). LDL was subjected to chromatography, and the column effluent was monitored at 280 nm. The retention time of the LDL peak was used to calculate the LDL diameter. We compared the HPGC method with gradient gel electrophoresis (GGE) on 2-10% nondenaturing polyacrylamide gels. In a group of 60 non-insulin-dependent diabetes mellitus patients, LDL size as measured by HPGC and GGE was highly correlated (r = 0.88, P <0.001). Good reproducibility, high precision, and the possibility of analyzing large series of samples are the main advantages of the automated HPGC method. Within-run and between-run CV for LDL size measured by HPGC were <0.1% and 0.2%, respectively. There was a significant inverse association between LDL size measured by HPGC and the logarithm of plasma triglycerides (r = -0.84, P <0.001), and a significant positive association with the LDL free cholesterol/protein ratio (r = 0.89, P <0.001). PMID- 9342013 TI - Early assessment of patients with suspected acute myocardial infarction by biochemical monitoring and neural network analysis. AB - Neural network analysis was applied for early diagnosis/exclusion of acute myocardial infarction (AMI), prediction of infarct size, and estimation of "time from onset of infarction." Eighty-eight patients admitted within 8 h after onset of chest pain were included. Blood samples for measurement of myoglobin, creatine kinase isoform MB, and troponin T were obtained every 30 min during the first 3 h and then after successively longer intervals. Data from 50 patients were used to train a set of neural network components of a decision support system. The performance of the system was evaluated and compared with experienced clinicians for the remaining 38 patients. The computer system detected myocardial infarction and predicted infarct size earlier than the clinicians, but did not differ significantly in terms of diagnostic sensitivity, specificity, and predictive values when disregarding time for diagnosis. With a cross-validation procedure the cumulated sensitivities of the computer system for the first five measurements were estimated to be (mean +/- 2SEM, n = 100): 0.77 +/- 0.03, 0.89 +/- 0.02, 0.94 +/- 0.02, 0.97 +/- 0.01, and 0.99 +/- 0.01, respectively, with corresponding cumulated specificities between 0.93 +/- 0.01 and 0.91 +/- 0.01. We concluded that neural network analysis of serial measurements of biochemical markers might provide useful support for the early assessment of patients with suspected AMI. PMID- 9342012 TI - Approach to maintaining comparability of biochemical data during long-term clinical trials. AB - Our objective was to design a structured approach to maintaining comparability of biochemical data during a long clinical trial. Maintaining the comparability of clinical and biochemical data obtained in long-term studies is essential, even though analytical methods in the laboratory may be changed, conventions on specimen handling and storage revised, calibration procedures updated, quality control systems replaced, and secular changes may occur. The United Kingdom Prospective Diabetes Study (UKPDS), a large randomized control trial investigating therapy for type 2 diabetes, was the setting for the study. Data were collected from 5102 subjects randomized since 1977. Our techniques included quality control, external quality assurance, direct comparison of laboratory methods when updating assays and statistical techniques for the detection of unsuspected changes in assay bias, laboratory comparisons of new with old assay methodologies, the realigning of data to current methods, the use of a suitable reference population for long-term monitoring, and rules to aid decision-making about clinical vs statistical significance. Procedures by which comparability of data is assured should be specified for all long-term trials and, where possible, comparison with reference methods should be detailed to allow results from different laboratories to be compared. PMID- 9342014 TI - Quality assessment of blood glucose testing in general practitioners' offices improves quality. AB - Measurement of blood glucose is a frequent test in Danish physicians' offices. We describe here a new design of external quality assessment wherein fresh, unstabilized whole-blood samples were analyzed by the physicians' office methods and by a hospital laboratory comparison method (glucose dehydrogenase assay, calibrated against NIST 909a). This approach was used in the offices of 171 general practitioners in our county during a 5-year period. The first survey, in 1992, revealed unsatisfactory performance in terms of our criterion that no difference between the physician's office and the hospital laboratory's single result should exceed +/-20% of the laboratory result. After initiation of a program of consultation and assistance, the proficiency testing rounds of 1994 and 1996 showed considerable improvement. Thus, whereas in 1992 12% of the values were outside the acceptance limits, in 1994 and 1996 the respective values were 4% and 3%. We believe the effect was mainly related to improvements in choice of technology, procedures, and handling of specimens. We conclude that the use of bedside instruments should be restricted to diagnosis of only severe hypo- or hyperglycemia and to monitoring glucose >5 mmo/L in already diagnosed diabetics. Our program differs from previous approaches to quality assessment and could also be useful for large hospitals' in-house proficiency testing. PMID- 9342015 TI - Kinetic assay of serum and urine for urea with use of urease and leucine dehydrogenase. AB - We describe a new kinetic assay for determining urea in serum or urine with use of urease (EC 3.5.1.5) and leucine dehydrogenase (EC 1.4.1.9). The latter enzyme is suitable for the kinetic assay of NH4+ because its Km value for NH4+ at pH 8.75 is large (approximately 500 mmol/L). Interference from endogenous NH4+ in serum or urine is obviated by subtraction of the assayed endogenous NH4+ value in a sample blank. For serum, within-assay CVs (n = 10) were 0.39-0.58%; day-to-day CVs (n = 10) were 1.56-2.30%. In urine, within-assay CVs (n = 10) were 0.86 1.15%. Analytical recovery of urea (0.893-71.4 mmol/L) added to patients' sera (urea 6.14 mmol/L) was 99.2-105.2%. The calibration curve for serum was linear through zero for urea concentrations up to 142.9 mmol/L and for urine up to 714.3 mmol/L. No influences of added ammonium ion, bilirubin, hemoglobin, ascorbic acid, or Intralipid were observed. The regression equations for this method (y) and conventional methods (x = Determiner-LUN for serum assays, Serotec UUR-R for urine) were: y = 1.016x - 0.12 mmol/L (r = 0.999, S(y/x) = 0.34 mmol/L, n = 100) for sera, and y = 1.070x - 12.6 mmol/L (r = 0.998, S(y/x) = 7.41 mmol/L, n = 100) for urine. PMID- 9342016 TI - Sensitive bioaffinity assays with individual microparticles and time-resolved fluorometry. AB - Future immunoassays and nucleic acid hybridization assays will be performed in miniaturized formats that utilize microchips or microparticles. This will require a sensitive detection technology that allows spatial resolution. By using fluorescent europium chelates and time-resolved microfluorometry, one can detect 11,000 europium molecules on individual microparticles. In a miniaturized noncompetitive immunoassay of prostate-specific antigen (PSA), we quantitatively detected 5 ng/L (0.05 amol per particle) of the analyte on an individual microparticle with excellent precision over the whole measurement range (CV <10%). Using a hybridization assay, we also could detect the deltaF508 mutation for cystic fibrosis on individual microparticles. Consequently, fluorescent lanthanide chelate labels and time-resolved microfluorometry qualify as the next generation of technology in this field. PMID- 9342017 TI - Candidate reference methods for hemoglobin A1c based on peptide mapping. AB - A reference method that specifically measures hemoglobin (Hb) A1c is an essential part of the reference system for the international standardization of Hb A1c/glycohemoglobin. We have developed a new method for quantification, based on the specific N-terminal residue of the hemoglobin beta-chains. Enzymatic cleavage of the intact hemoglobin molecule with endoproteinase Glu-C has been optimized to obtain the beta-N-terminal hexapeptides of Hb A1c and Hb A0. These peptides have been separated by reversed-phase HPLC and quantitated by electrospray ionization mass spectrometry (method A) or by capillary electrophoresis (method B). With these peptides and hyphenated separation techniques, it has been possible to overcome the insufficient resolution of currently used protein separation systems for Hb A1c. PMID- 9342018 TI - Assessment of monoethylglycinexylidide as measure of liver function for patients with chronic viral hepatitis. AB - The liver metabolizes lidocaine by oxidative deethylation to form monoethylglycinexylidide (MEGX), an analyte proposed as an index of liver function. We determined MEGX and lidocaine serum concentrations with the TDx (Abbott Laboratories) at baseline and 15, 30, 60, and 90 min after the intravenous administration of lidocaine (1 mg/kg), analyzing specimens from 12 apparently healthy volunteers and 40 patients with chronic viral hepatitis diagnosed by liver biopsy and serum tests. The patients were grouped on the basis of the histology activity index. The following laboratory tests were performed on serum specimens from all subjects: albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total bilirubin, and prothrombin time. The results showed no significant difference among the four groups for the concentrations of MEGX, lidocaine, and lidocaine/MEGX at the four time points. However, the concentrations of ALB, ALT, AST, AST/ALT, and prothrombin time were substantially different among the four groups. Thus, we conclude that assay of MEGX in our patients with chronic viral hepatitis did not contribute to the assessment of liver function when compared with apparently healthy volunteers and traditional tests of liver function. PMID- 9342019 TI - Significance of various parameters derived from biological variability of lipoprotein(a), homocysteine, cysteine, and total antioxidant status. AB - Analytical and biological components of variability and various derived indices have been determined for lipoprotein(a) [Lp(a)], homocysteine (Hcy), cysteine (Cys), and total antioxidant status (TAOS) in ostensibly healthy adult Caucasians and in stable outpatients with an increased serum Lp(a). In healthy Caucasians, average intraindividual biological CVs (CVb) were 20.0% for Lp(a), 9.4% for Hcy, 5.9% for Cys, and 2.8% for TAOS, CVbs being similar in men and women. In the outpatient group, CVbs were comparable for Hcy, Cys, and TAOS, but significantly lower for Lp(a) (7.5% vs 20.0%; P <0.0001). Moreover, a significant inverse relation between both biological and analytical CVs (CVa) and serum Lp(a) concentrations was demonstrated. We conclude that average CVa and CVb values, and hence average derived indices, are adequate for Hcy, Cys, and TAOS, whereas individual values should be used for Lp(a). PMID- 9342020 TI - Immunological measurement of transforming growth factor-beta 1 (TGF-beta1) in blood; assay development and comparison. AB - Development of a new, sensitive immunoassay for measuring transforming growth factor beta 1 (TGF-beta1) is described and compared with four commercially available TGF-beta1 immunoassays. Preanalytical conditions were evaluated. The nonlinearity found in serum or plasma is due to masking of TGF-beta1 by binding proteins in blood. Mixing TGF-beta1 with latency-associated peptide or alpha2 macroglobulin at physiological concentrations suppressed most of the TGF-beta1 signal. Plasma fibronectin showed no effect, even at concentrations exceeding its physiological range. Equilibrium concentrations computed from a model system confirmed the experimental results. Dilutional nonlinearity could be markedly reduced by an appropriately designed activation procedure that minimized the effects of reassociation between TGF-beta1 and its binding partners during restoration of a neutral pH. Plasma should be used for measuring TGF-beta1 in blood. Because serum TGF-beta1 is highly significantly correlated with the platelet count, probably most of the TGF-beta1 is released by platelet degranulation. PMID- 9342021 TI - HLA-B27 phenotyping with flow cytometry: further improvement by multiple monoclonal antibodies. AB - To establish the optimal flow cytometric method for HLA-B27 phenotyping, we compared several strategies, using three monoclonal anti-B27 antibodies (from the HLA-ABC-m3, GS145.2, and FD705 clones). We used a triple-color direct immunofluorescence assay, including a T-lymphocyte-specific antibody as an internal control and an anti-HLA-Bw4 antibody. Blood samples from >400 subjects were tested. From ROC curve analysis none of the three antibodies appeared to be suitable for use as a single typing reagent. The efficiency of the test was affected by cross-reactions with other HLA antigens, notably the HLA-B7 antigen. Preincubation with anti-B7 serum efficiently inhibited this cross-reaction and raised the test efficiency considerably. We concluded that none of the anti-B27 antibodies investigated is suitable for use as a single typing reagent. Additional typing of Bw4 is not valuable, whereas inhibition of cross-reactions due to the B7 antigen will considerably improve the performance of the test. We recommend that two different monoclonal anti-B27 antibodies be used for accurate and reliable HLA-B27 phenotyping with flow cytometry. PMID- 9342022 TI - Direct method for quantification of free malondialdehyde with high-performance capillary electrophoresis in biological samples. PMID- 9342023 TI - Oligonucleotide ligation assay for detection of apolipoprotein E polymorphisms. PMID- 9342024 TI - Measurement of Na+/K+-ATPase activity with an automated analyzer. PMID- 9342025 TI - Comparison of antinuclear antibody testing methods by ROC analysis with reference to disease diagnosis. PMID- 9342027 TI - Biological variation of superoxide dismutase in erythrocytes and glutathione peroxidase in whole blood. PMID- 9342026 TI - IMx tacrolimus II vs IMx tacrolimus microparticle enzyme immunoassay evaluated in renal and hepatic transplant patients. PMID- 9342028 TI - Measuring nitrous oxide in exhaled air by gas chromatography and infrared photoacoustic spectrometry. PMID- 9342029 TI - Extraction method and nonextracted kit method compared for measuring plasma aldosterone. PMID- 9342030 TI - Redox status of plasma homocysteine and related aminothiols in smoking and nonsmoking young adults. PMID- 9342031 TI - Urine organic acid profiling by capillary gas chromatography after a simple sample pretreatment. PMID- 9342032 TI - Rapid determination of total homocysteine in human plasma by using N(O,S) ethoxycarbonyl ethyl ester derivatives and gas chromatography-mass spectrometry. PMID- 9342033 TI - Ectopic production of creatine kinase MB: updated evaluation by mass assays. PMID- 9342034 TI - Acute bacterial prostatitis induces hematogenous dissemination of prostate epithelial cells. PMID- 9342035 TI - Misleading results with Opus Magnum hLH assay. PMID- 9342036 TI - Parameters relating to ultrasensitive prostate-specific antigen assays. PMID- 9342037 TI - Immunoassay of catecholamines and metabolites. PMID- 9342038 TI - Difference in hemoglobin-binding ability of polymers among haptoglobin phenotypes. PMID- 9342039 TI - Archenteron precursor cells can organize secondary axial structures in the sea urchin embryo. AB - Local cell-cell signals play a crucial role in establishing major tissue territories in early embryos. The sea urchin embryo is a useful model system for studying these interactions in deuterostomes. Previous studies showed that ectopically implanted micromeres from the 16-cell embryo can induce ectopic guts and additional skeletal elements in sea urchin embryos. Using a chimeric embryo approach, we show that implanted archenteron precursors differentiate autonomously to produce a correctly proportioned and patterned gut. In addition, the ectopically implanted presumptive archenteron tissue induces ectopic skeletal patterning sites within the ectoderm. The ectopic skeletal elements are bilaterally symmetric, and flank the ectopic archenteron, in some cases resulting in mirror-image, symmetric skeletal elements. Since the induced patterned ectoderm and supernumerary skeletal elements are derived from the host, the ectopic presumptive archenteron tissue can act to 'organize' ectopic axial structures in the sea urchin embryo. PMID- 9342040 TI - ladybird, a new component of the cardiogenic pathway in Drosophila required for diversification of heart precursors. AB - The embryonic heart precursors of Drosophila are arranged in a repeated pattern of segmental units. There is growing evidence that the development of individual elements of this pattern depends on both mesoderm intrinsic patterning information and inductive signals from the ectoderm. In this study, we demonstrate that two homeobox genes, ladybird early and ladybird late, are involved in the cardiogenic pathway in Drosophila. Their expression is specific to a subset of cardioblast and pericardial cell precursors and is critically dependent on mesodermal tinman function, epidermal Wingless signaling and the coordinate action of neurogenic genes. Negative regulation by hedgehog is required to restrict ladybird expression to two out of six cardioblasts in each hemisegment. Overexpression of ladybird causes a hyperplasia of heart precursors and alters the identity of even-skipped-positive pericardial cells. Loss of ladybird function leads to the opposite transformation, suggesting that ladybird participates in the determination of heart lineages and is required to specify the identities of subpopulations of heart cells. We find that both early Wingless signaling and ladybird-dependent late Wingless signaling are required for proper heart formation. Thus, we propose that ladybird plays a dual role in cardiogenesis: (i) during the early phase, it is involved in specification of a segmental subset of heart precursors as a component of the cardiogenic tinman cascade and (ii) during the late phase, it is needed for maintaining wingless activity and thereby sustaining the heart pattern process. These events result in a diversification of heart cell identities within each segment. PMID- 9342041 TI - The mouse Ulnaless mutation deregulates posterior HoxD gene expression and alters appendicular patterning. AB - The semi-dominant mouse mutation Ulnaless alters patterning of the appendicular but not the axial skeleton. Ulnaless forelimbs and hindlimbs have severe reductions of the proximal limb and less severe reductions of the distal limb. Genetic and physical mapping has failed to separate the Ulnaless locus from the HoxD gene cluster (Peichel, C. L., Abbott, C. M. and Vogt, T. F. (1996) Genetics 144, 1757-1767). The Ulnaless limb phenotypes are not recapitulated by targeted mutations in any single HoxD gene, suggesting that Ulnaless may be a gain-of function mutation in a coding sequence or a regulatory mutation. Deregulation of 5' HoxD gene expression is observed in Ulnaless limb buds. There is ectopic expression of Hoxd-13 and Hoxd-12 in the proximal limb and reduction of Hoxd-13, Hoxd-12 and Hoxd-11 expression in the distal limb. Skeletal reductions in the proximal limb may be a consequence of posterior prevalence, whereby proximal misexpression of Hoxd-13 and Hoxd-12 results in the transcriptional and/or functional inactivation of Hox group 11 genes. The Ulnaless digit phenotypes are attributed to a reduction in the distal expression of Hoxd-13, Hoxd-12, Hoxd-11 and Hoxa-13. In addition, Hoxd-13 expression is reduced in the genital bud, consistent with the observed alterations of the Ulnaless penian bone. No alterations of HoxD expression or skeletal phenotypes were observed in the Ulnaless primary axis. We propose that the Ulnaless mutation alters a cis-acting element that regulates HoxD expression specifically in the appendicular axes of the embryo. PMID- 9342042 TI - Ulnaless (Ul), a regulatory mutation inducing both loss-of-function and gain-of function of posterior Hoxd genes. AB - Ulnaless (Ul), an X-ray-induced dominant mutation in mice, severely disrupts development of forearms and forelegs. The mutation maps on chromosome 2, tightly linked to the HoxD complex, a cluster of regulatory genes required for proper morphogenesis. In particular, 5'-located (posterior) Hoxd genes are involved in limb development and combined mutations within these genes result in severe alterations in appendicular skeleton. We have used several engineered alleles of the HoxD complex to genetically assess the potential linkage between these two loci. We present evidence indicating that Ulnaless is allelic to Hoxd genes. Important modifications in the expression patterns of the posterior Hoxd-12 and Hoxd-13 genes at the Ul locus suggest that Ul is a regulatory mutation that interferes with a control mechanism shared by multiple genes to coordinate Hoxd function during limb morphogenesis. PMID- 9342043 TI - Role of GGF/neuregulin signaling in interactions between migrating neurons and radial glia in the developing cerebral cortex. AB - During neuronal migration to the developing cerebral cortex, neurons regulate radial glial cell function and radial glial cells, in turn, support neuronal cell migration and differentiation. To study how migrating neurons and radial glial cells influence each others' function in the developing cerebral cortex, we examined the role of glial growth factor (a soluble form of neuregulin), in neuron-radial glial interactions. Here, we show that GGF is expressed by migrating cortical neurons and promotes their migration along radial glial fibers. Concurrently, GGF also promotes the maintenance and elongation of radial glial cells, which are essential for guiding neuronal migration to the cortex. In the absence of GGF signaling via erbB2 receptors, radial glial development is abnormal. Furthermore, GGF's regulation of radial glial development is mediated in part by brain lipid-binding protein (BLBP), a neuronally induced, radial glial molecule, previously shown to be essential for the establishment and maintenance of radial glial fiber system. The ability of GGF to influence both neuronal migration and radial glial development in a mutually dependent manner suggests that it functions as a mediator of interactions between migrating neurons and radial glial cells in the developing cerebral cortex. PMID- 9342044 TI - Synergistic interactions between bFGF and a TGF-beta family member may mediate myogenic signals from the neural tube. AB - Development of the myotome within somites depends on unknown signals from the neural tube. The present study tested the ability of basic fibroblast growth factor (bFGF), transforming growth factor-beta1 (TGF-beta1) and dorsalin-1 (dsl 1) to promote myogenesis in stage 10-14 chick paraxial mesoderm utilizing 72 hour explant cultures. Each of these factors alone and the combination of bFGF with dsl-1 had limited to no myogenic-promoting activity, but the combination of bFGF with TGF-beta1 demonstrated a potent dose-dependent effect. In addition, bFGF enhanced the survival/proliferation of somite cells. 98% of stage 10-11 caudal segmental plate explants treated with bFGF plus TGF-beta1, exhibited myosin heavy chain (MHC)-positive cells (avg.=60 per explant), whereas only 15% of similarly treated somites responded with an average of 5 MHC-positive cells. Thus at stage 10-11, there are rostrocaudal differences in myogenic responsiveness with the caudal (more 'immature') paraxial mesoderm being more myogenically responsive to these factors than are somites. It was also discovered that 17% of stage 10-11 caudal segmental plate explants exhibited several MHC-positive cells even when cultured without added growth factors, further demonstrating a different myogenic potential of the caudal paraxial mesoderm. Stage 13-14 paraxial mesoderm also exhibited a myogenic response to bFGF/TGF-beta1 but, unlike stage 10-11 embryos, both somites and segmental plate exhibited a strong response. A two-step mechanism for the bFGF/TGF-beta1 effect is suggested by the finding that only TGF beta1 was required during the first 12 hours of culture, whereas bFGF plus a TGF beta-like factor were required for the remainder of the culture. The biological relevance of the findings with bFGF is underscored by the observation that a monoclonal antibody to bFGF inhibited myogenic signaling from the dorsal neural tube. However, a monoclonal antibody that can neutralize the three factors TGF beta1, TGF-beta2 and TGF-beta3 did not block myogenic signals from the neural tube, raising the possibility that another TGF-beta family member may be involved in vivo. PMID- 9342045 TI - Cell migration in the developing chick diencephalon. AB - We previously reported that retrovirally marked clones in the mature chick diencephalon were widely dispersed in the mediolateral, dorsoventral and rostrocaudal planes. The current study was undertaken to define the migration routes that led to the dispersion. Embryos were infected between stages 10 and 14 with a retroviral stock encoding alkaline phosphatase and a library of molecular tags. Embryos were harvested 2.5-5.5 days later and the brains were fixed and serially sectioned. Sibling relationships were determined following PCR amplification and sequencing of the molecular tag. On embryonic day 4, all clones were organized in radial columns spanning the neuroepithelium, which was composed primarily of a ventricular zone at this age. No tangential migration was seen in the ventricular zone. On embryonic day 5, most clones remained radial with many cells located in the ventricular zone; however, a few clones had cells migrating perpendicular to the radial column, in either a rostrocaudal or dorsoventral direction. The tangential migration began just beyond the basal limit of the ventricular zone. On embryonic days 6 and 7, many clones had cells migrating perpendicular to the radial column, which spanned from the ventricular to the pial surface. The migrating cells appeared to be aligned along axes that were perpendicular to the radial column. Using a combination of DiI tracing, immunohistochemistry and electron microscopy, we have determined that axonal tracts are present and are aligned with the migrating cells, suggesting that they support the non-radial cell migration. These data indicate that migration along pathways independent of radial glia occur outside of the ventricular zone in more than 50% of the clones in the chick diencephalon. PMID- 9342046 TI - MAP kinase in situ activation atlas during Drosophila embryogenesis. AB - Receptor tyrosine kinases (RTKs) and the signaling cascades that they trigger play central roles in diverse developmental processes. We describe the capacity to follow the active state of these signaling pathways in situ. This is achieved by monitoring, with a specific monoclonal antibody, the distribution of the active, dual phosphorylated form of MAP kinase (ERK). A dynamic pattern is observed during embryonic and larval phases of Drosophila development, which can be attributed, to a large extent, to the known RTKs. This specific detection has enabled us to determine the time of receptor activation, visualize gradients and boundaries of activation, and postulate the distribution of active ligands. Since the antibody was raised against the phosphorylated form of a conserved ERK peptide containing the TEY motif, this approach is applicable to a wide spectrum of multicellular organisms. PMID- 9342047 TI - The Drosophila gene morula inhibits mitotic functions in the endo cell cycle and the mitotic cell cycle. AB - In the endo cell cycle, rounds of DNA replication occur in the absence of mitosis, giving rise to polyploid or polytene cells. We show that the Drosophila morula gene is essential to maintain the absence of mitosis during the endo cycle. During oogenesis in wild-type Drosophila, nurse cells become polyploid and do not contain cyclin B protein. Nurse cells in female-sterile alleles of morula begin to become polyploid but revert to a mitotic-like state, condensing the chromosomes and forming spindles. In strong, larval lethal alleles of morula, the polytene ring gland cells also inappropriately regress into mitosis and form spindles. In addition to its role in the endo cycle, morula function is necessary for dividing cells to exit mitosis. Embryonic S-M cycles and the archetypal (G1-S G2-M) cell cycle are both arrested in metaphase in different morula mutants. These phenotypes suggest that morula acts to block mitosis-promoting activity in both the endo cycle and at the metaphase/anaphase transition of the mitotic cycle. Consistent with this, we found cyclin B protein to be inappropriately present in morula mutant nurse cells. Thus morula serves a dual function as a cell cycle regulator that promotes exit from mitosis and maintains the absence of mitosis during the endo cycle, possibly by activating the cyclin destruction machinery. PMID- 9342048 TI - Cell cycle progression, growth and patterning in imaginal discs despite inhibition of cell division after inactivation of Drosophila Cdc2 kinase. AB - During larval development, Drosophila imaginal discs increase in size about 1000 fold and cells are instructed to acquire distinct fates as a function of their position. The secreted signaling molecules Wingless and Decapentaplegic have been implicated as sources of positional information that globally control growth and patterning. Evidence has also been presented that local cell interactions play an important role in controlling cell proliferation in imaginal discs. As a first step to understanding how patterning cues influence growth we investigated the effects of blocking cell division at different times and in spatially controlled manner by inactivation of the mitotic kinase Cdc2 in developing imaginal discs. We find that cell growth continues after inactivation of Cdc2, with little effect on overall patterning. The mechanisms that regulate size of the disc therefore do not function by regulating cell division, but appear to act primarily by regulating size in terms of physical distance or tissue volume. PMID- 9342049 TI - The Drosophila sugarless gene modulates Wingless signaling and encodes an enzyme involved in polysaccharide biosynthesis. AB - We have identified and characterized a Drosophila gene, which we have named sugarless, that encodes a homologue of vertebrate UDP-glucose dehydrogenase. This enzyme is essential for the biosynthesis of various proteoglycans, and we find that in the absence of both maternal and zygotic activities of this gene, mutant embryos develop with segment polarity phenotypes reminiscent to loss of either Wingless or Hedgehog signaling. To analyze the function of Sugarless in cell-cell interaction processes, we have focused our analysis on its requirement for Wingless signaling in different tissues. We report that sugarless mutations impair signaling by Wingless, suggesting that proteoglycans contribute to the reception of Wingless. We demonstrate that overexpression of Wingless can bypass the requirement for sugarless, suggesting that proteoglycans modulate signaling by Wingless, possibly by limiting its diffusion and thereby facilitating the binding of Wingless to its receptor. We discuss the possibility that tissue specific regulation of proteoglycans may be involved in regulating both Wingless short- or long-range effects. PMID- 9342050 TI - Isoform-specific expression and function of neuregulin. AB - Neuregulin (also known as NDF, heregulin, ARIA, GGF or SMDF), induces cell growth and differentiation. Biological effects of neuregulin are mediated by members of the erbB family of tyrosine kinase receptors. Three major neuregulin isoforms are produced from the gene, which differ substantially in sequence and in overall structure. Here we use in situ hybridization with isoform-specific probes to illustrate the spatially distinct patterns of expression of the isoforms during mouse development. Ablation of the neuregulin gene in the mouse has demonstrated multiple and independent functions of this factor in development of both the nervous system and the heart. We show here that targeted mutations that affect different isoforms result in distinct phenotypes, demonstrating that isoforms can take over specific functions in vivo. Type I neuregulin is required for generation of neural crest-derived neurons in cranial ganglia and for trabeculation of the heart ventricle, whereas type III neuregulin plays an important role in the early development of Schwann cells. The complexity of neuregulin functions in development is therefore due to independent roles played by distinct isoforms. PMID- 9342051 TI - Nitric oxide, an endogenous regulator of Dictyostelium discoideum differentiation. AB - We have previously demonstrated that nitric oxide (NO)-generating compounds inhibit D. discoideum differentiation by preventing the initiation of cAMP pulses (Tao, Y., Howlett, A. and Klein, C. (1996) Cell. Signal. 8, 37-43). In the present study, we demonstrate that cells produce NO at a relatively constant rate during the initial phase of their developmental cycle. The addition of oxyhemoglobin, an NO scavenger, stimulates cell aggregation, suggesting that NO has a negative effect on the development of aggregation competence. Starvation of cells in the presence of glucose, which has been shown to prevent the initiation of cAMP pulses (Darmon, M. and Klein, C. (1978) Dev. Biol. 63, 377-389), results in an increased production of NO. The inhibition of cell aggregation by glucose treatment can be reversed by oxyhemoglobin. These findings indicate that NO is a signaling molecule for D. discoideum cells and that physiological or environmental conditions that enhance external NO levels will delay the initiation of cAMP pulses, which are essential for cell differentiation. PMID- 9342052 TI - Transmembrane grasshopper Semaphorin I promotes axon outgrowth in vivo. AB - Members of the Semaphorin family of glycoproteins play an important role in axonal pathfinding by functioning as inhibitory guidance cues. Here we provide evidence that a transmembrane form of Semaphorin (Semaphorin I), which is expressed by bands of epithelial cells in the developing grasshopper limb bud, functions as an attractive/permissive cue for the growth cones of the subgenual organ. In addition, we demonstrate that Semaphorin I is needed for initial axonal outgrowth from the subgenual organ. These results are consistent with an alternative function for a transmembrane form of Semaphorin and may explain the previously reported arrest of the proximal extension of the subgenual organ growth cones in the absence of the Ti1 pioneer pathway. PMID- 9342053 TI - A forkhead gene related to HNF-3beta is required for gastrulation and axis formation in the ascidian embryo. AB - We have isolated a member of the HNF-3/forkhead gene family in ascidians as a means to determine the role of winged-helix genes in chordate development. The MocuFH1 gene, isolated from a Molgula oculata cDNA library, exhibits a forkhead DNA-binding domain most similar to zebrafish axial and rodent HNF-3beta. MocuFH1 is a single copy gene but there is at least one other related forkhead gene in the M. oculata genome. The MocuFH1 gene is expressed in the presumptive endoderm, mesenchyme and notochord cells beginning during the late cleavage stages. During gastrulation, MocuFH1 expression occurs in the prospective endoderm cells, which invaginate at the vegetal pole, and in the presumptive notochord and mesenchyme cells, which involute over the anterior and lateral lips of the blastopore, respectively. However, this gene is not expressed in the presumptive muscle cells, which involute over the posterior lip of the blastopore. MocuFH1 expression continues in the same cell lineages during neurulation and axis formation, however, during the tailbud stage, MocuFH1 is also expressed in ventral cells of the brain and spinal cord. The functional role of the MocuFH1 gene was studied using antisense oligodeoxynucleotides (ODNs), which transiently reduce MocuFH1 transcript levels during gastrulation. Embryos treated with antisense ODNs cleave normally and initiate gastrulation. However, gastrulation is incomplete, some of the endoderm and notochord cells do not enter the embryo and undergo subsequent movements, and axis formation is abnormal. In contrast, the prospective muscle cells, which do not express MocuFH1, undergo involution and later express muscle actin and acetylcholinesterase, markers of muscle cell differentiation. The results suggest that MocuFH1 is required for morphogenetic movements of the endoderm and notochord precursor cells during gastrulation and axis formation. The effects of inhibiting MocuFH1 expression on embryonic axis formation in ascidians are similar to those reported for knockout mutations of HNF-3beta in the mouse, suggesting that HNF-3/forkhead genes have an ancient and fundamental role in organizing the body plan in chordates. PMID- 9342054 TI - Imprinting of Igf2 and H19 from a 130 kb YAC transgene. AB - A stringent test for imprint control elements is to examine their function at ectopic loci in transgenic experiments. Igf2 and H19 are part of a larger imprinting region and as a first step, we examined these reciprocally imprinted genes in transgenic experiments using a 130 kb YAC clone. After paternal inheritance, H19 was appropriately repressed and Igf2 was expressed, irrespective of copy number or genetic background. After maternal inheritance H19 was consistently expressed, albeit with some variability. The levels of H19 expression per copy of the transgene inversely correlated with Igf2 (-lacZ) expression in cis. The consistent imprinting of H19 from this YAC contrasts with the previously described imprinting of mini-H19 transgenes, which only occurs at multi-copy loci, is inconsistent, and is prone to genetic background effects. We propose a novel model in which silencing of the H19 gene is the default state and its activation after maternal inheritance is the key mechanistic event for imprinting in this region. In addition, in situ analysis of the Igf2-lacZ reporter indicates that additional mesoderm-specific enhancers are present within the YAC clone. No obvious phenotype was detected from the excess gene dosage of H19. PMID- 9342055 TI - Macrophages induce apoptosis in normal cells in vivo. AB - It is well established that macrophages have a function in scavenging apoptotic bodies from cells undergoing programmed cell death. Here we show that macrophages can also induce apoptosis of normal cells. Using injected toxic liposomes to eliminate macrophages in the anterior chamber of the rat eye, we provide direct evidence that, in vivo, macrophages induce apoptosis in normal vascular endothelial cells during programmed capillary regression. Macrophage elimination resulted in the survival of endothelial cells that normally would die and the persistence of functional capillaries. Furthermore, replacement of eliminated macrophages with bone-marrow-derived macrophages 'rescued' lack of capillary regression. Viability of the persistent target cells was demonstrated through their lack of apoptotic morphology, expression of intracellular esterases and synthesis of DNA. These results uncover a new function for macrophages in the remodeling of tissues through the induction of programmed cell death and provide direct evidence of a key role for macrophages in capillary regression. PMID- 9342056 TI - Genetic control of brain morphogenesis through Otx gene dosage requirement. AB - Understanding the genetic mechanisms that control patterning of the vertebrate brain represents a major challenge for developmental neurobiology. Previous data suggest that Otx1 and Otx2, two murine homologs of the Drosophila orthodenticle (otd) gene, might both contribute to brain morphogenesis. To gain insight into this possibility, the level of OTX proteins was modified by altering in vivo the Otx gene dosage. Here we report that Otx genes may cooperate in brain morphogenesis and that a minimal level of OTX proteins, corresponding either to one copy each of Otx1 and Otx2, or to only two copies of Otx2, is required for proper regionalization and subsequent patterning of the developing brain. Thus, as revealed by anatomical and molecular analyses, only Otx1-/-; Otx2+/- embryos lacked mesencephalon, pretectal area, dorsal thalamus and showed an heavy reduction of the Ammon's horn, while the metencephalon was dramatically enlarged occupying the mesencencephalic area. In 8.5 days post coitum (d.p.c.) Otx1-/-; Otx2+/- embryos, the expression patterns of mesencephalic-metencephalic (mes-met) markers such as En-1 and Wnt-1 confirmed the early presence of the area fated to give rise to mesencephalon and metencephalon while Fgf-8 transcripts were improperly localized in a broader domain. Thus, in Otx1-/-; Otx2+/- embryos, Fgf 8 misexpression is likely to be the consequence of a reduced level of specification between mes-met primitive neuroepithelia that triggers the following repatterning involving the transformation of mesencephalon into metencephalon, the establishment of an isthmic-like structure in the caudal diencephalon and, by 12.5 d.p.c., the telencephalic expression of Wnt-1 and En-2. Taken together these findings support the existence of a molecular mechanism depending on a precise threshold of OTX proteins that is required to specify early regional diversity between adjacent mes-met territories and, in turn, to allow the correct positioning of the isthmic organizer. PMID- 9342057 TI - Ectopic expression of the Drosophila Bam protein eliminates oogenic germline stem cells. AB - The Drosophila germ-cell lineage has emerged as a remarkable system for identifying genes required for changes in cell fate from stem cells into more specialized cells. Previous work indicates that bam expression is necessary for cystoblast differentiation; bam mutant germ cells fail to differentiate, but instead proliferate like stem cells. This paper reports that ectopic expression of bam is sufficient to extinguish stem cell divisions. Heat-induced bam+ expression specifically eliminated oogenic stem cells while somatic stem cell populations were not affected. Together with previous studies of the timing of bam mRNA and protein expression and the state of arrest in bam mutant cells, these data implicate Bam as a direct regulator of the switch from stem cell to cystoblast. Surprisingly, ectopic bam+ had no deleterious consequences for male germline cells suggesting that Bam may regulate somewhat different steps of germ cell development in oogenesis and spermatogenesis. We discuss a model for how bam+ could direct differentiation based on our data (McKearin and Ohlstein, 1995) that Bam protein is essential to assemble part of the germ-cell-specific organelle, the fusome. We propose that fusome biogenesis is an obligate step for cystoblast cell fate and that Bam is the limiting factor for fusome maturation in female germ cells. PMID- 9342058 TI - cut interacts with Notch and protein kinase A to regulate egg chamber formation and to maintain germline cyst integrity during Drosophila oogenesis. AB - Communications between the germline and the soma during Drosophila oogenesis have been previously shown to be essential for the formation of egg chambers and to establish polarity in the developing oocyte. In this report, we demonstrate that the function of a somatically expressed gene, cut, is critical for maintaining the structural integrity of germline-derived cells and their arrangement within an egg chamber. Genetic manipulations of cut activity resulted in defective packaging of germline-derived cysts into egg chambers and disintegration of the structural organization of oocyte-nurse cell complexes to generate multinucleate germline-derived cells. We also found that cut interacts genetically with the Notch gene and with the catalytic subunit of Protein kinase A gene during egg chamber morphogenesis. Since cut expression is restricted to the somatic follicle cells and cut mutant germline clones are phenotypically normal, we propose that the defects in the assembly of egg chambers and the changes in germline cell morphology observed in cut mutant egg chambers are the result of altered interactions between follicle cells and germline cells. cut encodes a nuclear protein containing DNA-binding motifs, and we suggest that it participates in intercellular communications by regulating the expression of molecules that directly participate in this process. PMID- 9342059 TI - The associations between cervicovaginal HIV shedding, sexually transmitted diseases and immunosuppression in female sex workers in Abidjan, Cote d'Ivoire. AB - OBJECTIVE: To measure the frequency and associated factors of cervicovaginal HIV shedding and to determine the impact of sexually transmitted disease (STD) treatment on HIV shedding. DESIGN: Cross-sectional study with 1-week follow-up. SETTING: Confidential clinic for female sex workers in Abidjan, Cote d'Ivoire. PARTICIPANTS: A total of 1201 female sex workers. INTERVENTIONS: STD treatment based on clinical signs. MAIN OUTCOME MEASURES: HIV serostatus; cervicovaginal HIV shedding at enrollment and at 1-week follow-up; STD status at enrollment and at 1-week follow-up. RESULTS: Cervicovaginal shedding of HIV-1 in HIV-1 seropositive women was more frequent (96 out of 404, 24%) than shedding of HIV-2 in HIV-2-seropositive women [one out of 21, 5%; odds ratio (OR), 6.2; 95% confidence interval (CI), 1.0-261]. Among 609 HIV-1-seropositive or dually seroreactive women, HIV-1 shedding was significantly more frequent in immunosuppressed women [adjusted OR (AOR), 6.3; 95% CI, 3.4-11.9; and AOR, 2.9; 95% CI, 1.6-5.0 for CD4 < 14% and CD4 14-28%, respectively, versus CD4 > 28%], and in women with Neisseria gonorrhoeae (AOR, 1.9; 95% CI, 1.2-3.0), those with Chlamydia trachomatis (AOR, 2.5; 95% CI, 1.1-5.8), and with a cervical or vaginal ulcer (AOR, 3.9; 95% CI, 2.1-7.4). HIV-1 shedding decreased from 42 to 21% (P < 0.005) in women whose STD were cured. CONCLUSIONS: These data help to explain the difference in transmissibility between HIV-1 and HIV-2 and the increased infectiousness of HIV in the presence of immunosuppression and STD. In addition, they lend biological plausibility to arguments for making STD control an integral part of HIV prevention strategies in Africa. PMID- 9342060 TI - Toxicity, efficacy, plasma drug concentrations and protease mutations in patients with advanced HIV infection treated with ritonavir plus saquinavir. Swiss HIV Cohort Study. AB - OBJECTIVE: To assess the safety, efficacy and plasma drug levels of the combination of ritonavir plus saquinavir for the treatment of advanced HIV infection. DESIGN: Multicentre pilot study. PATIENTS: Eighteen protease inhibitor naive patients, with intolerance or contraindication to reverse transcriptase inhibitors, a median CD4 cell count of 12 x 10(6)/l (range, 1-50 x 10(6)/l), and a median HIV viraemia of 5.25 log10 copies/ml (range, 4.00-6.13 log10 copies/ml). METHODS: Patients received 600 mg twice daily of both ritonavir and saquinavir. Viraemia was measured at baseline and at weeks 5, 9 and 13. Response was defined as a drop of viraemia of more than 1 log10 at week 5. Plasma drug levels were determined after at least 3 weeks of combined treatment: samples were collected before and 1, 2, and 4 h after the morning ingestion of both drugs. The protease gene was sequenced at baseline and under treatment. RESULTS: Among the 16 patients evaluable at week 5, 11 were responders, and among these patients, six remained responders at week 13 (two with undetectable viraemia). Study discontinuations were due to side-effects (n = 4), patient choice (n = 3), protocol violation (n = 1) and death (n = 1). Responders had higher drug levels than non-responders (P < 0.01 for saquinavir, P = 0.04 for ritonavir). In two non responders, development of multiple new mutations at positions 10, 20, 48, 82, 84 and 90 was observed after 5-13 weeks. CONCLUSION: The response to ritonavir plus saquinavir in advanced HIV infection is unpredictable. A minority of patients respond with disappearance of HIV viraemia. In other patients, rapid cumulative emergence of protease mutations conferring resistance to treatment cannot always be prevented by good compliance and relatively high plasma drug levels. PMID- 9342061 TI - Impact of protease inhibitors on AIDS-defining events and hospitalizations in 10 French AIDS reference centres. Federation National des Centres de Lutte contre le SIDA. AB - OBJECTIVE: To assess the clinical and economic consequences of the use of protease inhibitors in the treatment of HIV infection. DESIGN: Multicentric, observational, retrospective cohort study. SETTING: Ten AIDS reference centres in France. PATIENTS: All patients followed in each centre from September 1995 through October 1996. MAIN OUTCOME MEASURES: AIDS-defining events, death, health care resources use, administration of antiretroviral therapy. RESULTS: Data from 7749 patients in 10 centres showed a drop in hospitalization days by 35%, new AIDS cases by 35%, and deaths by 46%. In the same period, the proportion of patients receiving antiretrovirals rose from 36 to 53% including highly active antiretroviral therapy (HAART), which rose from 0.3 to 18%. Overall cost evaluation showed a slight increase of monthly treatment cost of US$ 12 per patient. Comparison of the three centres that used HAART earliest to the three centres that used it latest showed a clear benefit to early HAART with a drop in hospitalization days by 41%, new AIDS cases by 41% and deaths by 69%. The proportion of patients with HAART rose to 27% and monthly health-care cost decreased by US$ 248852 (i.e., by US$ 101 per patient per month). Late prescribing centres experienced a less marked effect with a drop in hospitalization days by 22%, new AIDS cases by 31%, and deaths by 32.5%. Proportion of patients with HAART rose to 12% and monthly health-care costs increased by US$ 113578 (i.e., by US$ 38 per patient per month). CONCLUSIONS: This study supports the extensive use of HAART in HIV-infected patients. PMID- 9342062 TI - Protease inhibitor therapy in children with perinatally acquired HIV infection. AB - OBJECTIVE: To review the short-term response and safety of protease inhibitor therapy in HIV-infected children. DESIGN: Retrospective chart review of open label protease inhibitor-containing combination therapy. SETTING: Two urban pediatric HIV centers. PATIENTS: Twenty-eight HIV-infected children were prescribed 30 protease inhibitor-containing antiretroviral therapy combinations. The median age at initiation of protease inhibitor antiretroviral therapy was 79 months. Patients had been on previous antiretroviral therapy for a mean of 45.5 months. RESULTS: Of the 28 children who completed at least 1 month of therapy, 26 experienced marked virologic and immunologic improvement (mean maximal decrease in viral load 1.90 log10 copies/ml; SD, 0.8; mean maximal rise in CD4+ lymphocytes of 279 x 10(6)/l; SD, 300 x 10(6)/l). Eleven patients achieved a viral nadir of < 400 copies/ml, and seven sustained this level of viral suppression for a mean of 6 months. Indinavir use was associated with a high incidence of renal side-effects, including two patients who developed interstitial nephritis. Two patients on ritonavir experienced a significant elevation of liver enzymes. CONCLUSIONS: Protease inhibitor therapy was associated with substantial short-term virologic and immunologic improvement in this primarily heavily pretreated cohort, with 25% maintaining a viral load of < 400 copies/ml after 6 months of therapy. There was a significant rate of adverse events. Pharmacokinetic and safety data are needed to guide aggressive antiretroviral therapy in HIV-infected children, and further treatment options are required for those failing or intolerant to the available protease inhibitors. PMID- 9342063 TI - The influence of MT-2 tropism on the prognostic implications of the delta32 deletion in the CCR-5 gene. AB - BACKGROUND: Long-term non-progression in HIV-1-infected patients has been reported to be associated with a 32 base-pair deletion (delta32) in one CCR-5 allele. The normal gene product acts as a coreceptor for HIV cell entry and is essential for infection of cells by non-syncytium-inducing and MT-2-negative HIV 1 strains. METHODS: Forty individuals were studied, all of whom had been HIV-1 seropositive for a mean of 8 years. RESULTS: Eight (20%) were heterozygous for the CCR-5 allele delta32 deletion. Six of these eight patients harboured MT-2 negative HIV-1 strains. Of these six, three were long-term non-progressors with a positive CD4 cell slope, not receiving antiretroviral treatment, whereas the other three were progressors (mean CD4 cell decline, 3.8 x 10(6)/l per month) receiving antiretroviral combination therapy. Two of the eight patients with the delta32 deletion had MT-2-positive HIV-1 strains. Both had very rapid CD4 cell decline (6.7 and 7.6 x 10(6)/l per month, respectively), despite triple antiretroviral therapy including a protease inhibitor. One of the patients with an MT-2-positive virus strain has suffered from Pneumocystis carinii bronchitis and the other from cytomegalovirus colitis. CONCLUSIONS: Disease progression may also occur in individuals with the coreceptor deficiency, especially in association with MT-2-positive HIV-1 strains. It is suggested that MT-2-positive HIV-1 enters cells through the CXC chemokine receptor-4 fusin coreceptor, thus circumventing the defective CC chemokine receptor-5 coreceptor. Various levels of expression of the wild-type CCR-5 gene and the gene with the delta32 deletion might explain variations in the disease progression in heterozygous patients with MT-2-negative HIV-1 strains. PMID- 9342064 TI - HIV-1 Tat protein exits from cells via a leaderless secretory pathway and binds to extracellular matrix-associated heparan sulfate proteoglycans through its basic region. AB - OBJECTIVE: To analyze the mechanisms of release and the extracellular fate of the HIV-1 Tat protein and to determine the Tat domain binding to the extracellular matrix. DESIGN AND METHODS: Release of Tat was studied by pulse-chase experiments with Tat-transfected COS-1 cells in the presence or absence of different serum concentrations, temperatures and drugs inhibiting the classical secretion pathway or endo-exocytosis, such as brefeldin A and methylamine. The binding of extracellular Tat to heparan sulfate proteoglycans (HSPG) was determined by using trypsin, heparin or heparinase in pulse-chase experiments, by gel shift and competition assays with radiolabeled heparin, and by heparin-affinity chromatography. The mapping of the Tat binding site to heparin was defined by functional assays of rescue of Tat-defective HIV-1 proviruses. RESULTS: Tat is released in the absence of cell death or permeability changes. Tat release is dependent upon the temperature and serum concentration, and it is not blocked by brefeldin A or methylamine. After release, a portion of the protein remains in a soluble form whereas the other binds to extracellular matrix (ECM)-associated HSPG. The HSPG-bound Tat can be retrieved into a soluble form by heparin, heparinase or trypsin. Binding to heparin is competed out by heparin-binding factors such as basic fibroblast growth factor (bFGF), and it is mediated by the Tat basic region which forms a specific complex with heparin which blocks HIV-1 rescue by exogenous Tat and allows purification of a highly biologically active protein. CONCLUSIONS: These results demonstrate that Tat exits from intact cells through a leaderless secretion pathway which shares several features with that of acid FGF or bFGF. The released Tat binds to HSPG through its basic region and this determines its storage into the ECM, as occurs for bFGF. PMID- 9342065 TI - Development of genetic vaccines for pathogenic genes: construction of attenuated vif DNA immunization cassettes. AB - OBJECTIVE: To develop a putative immunization cassette using HIV-1 vif accessory gene derived from HIV-1 clinical specimens as a component of a DNA vaccine for HIV-1. METHODS: vif genes were cloned from HIV-1-infected patients and the sequence variation present within the patients was analyzed. Prototypic genetic variants were selected and the ability of these clones to induce humoral and cellular immune responses was studied in animals. The selected protective genetic variants were biologically characterized through transcomplementation assays using primary cells infected with a vif-defective HIV-1 proviral clone. RESULTS: Analysis of vif variants from different patients revealed that vif is highly conserved with the open reading frame remaining intact in vivo. It was shown that attenuated vif clones from HIV-1-infected subjects can effectively induce both humoral and cellular responses against Vif protein in mice. Evaluation of the cellular responses in vitro using human cellular targets infected with a clinical HIV-1 isolate showed that vif clones could induce cellular responses capable of destroying the virus. CONCLUSIONS: The vif variants developed in this study exhibited non-productive phenotypes, yet were capable of inducing specific immune responses against HIV-1. These constructs could be used as part of a DNA vaccine strategy for HIV-1. This vaccine adaptation strategy could be used for the development of immunogens for any pathogen resulting in cross-reactive immunity and attenuated gene pathogenesis. PMID- 9342066 TI - HIV infection alters the production of both type 1 and 2 cytokines but does not induce a polarized type 1 or 2 state. AB - OBJECTIVE: To test the T-helper (TH)1/TH2 cytokine paradigm in HIV infection. DESIGN AND METHODS: Cytokine profiles in two separate studies of HIV patients and controls are presented: (i) a longitudinal study of HIV patients with CD4 counts > 500 x 10(6)/l tested at three timepoints compared with controls; (ii) a blinded cross-sectional study of controls and patients with high (> 500 x 10(6)/l) and low (< 500 x 10(6)/l) CD4 counts. Peripheral blood mononuclear cells (PBMC) from patients and controls were tested for the production of two type 1 [interleukin (IL)-2, interferon (IFN)-gamma] and two type 2 (IL-4, IL-10) cytokines by enzyme linked immunosorbent assay. Both spontaneous and mitogen-induced cytokine production was measured. RESULTS: HIV infection was noted to have the following effects on cytokine production: (i) it led to the in vivo activation of type 2 cytokines in a small group of individuals with high CD4 numbers characterized by the spontaneous release of IL-4 and IL-10. Longitudinal data showed high spontaneous IL-4 and IL-10 to be a consistent feature of the patient group (at each timepoint some patients were high producers) but to be variable in a given individual; (ii) HIV infection impaired the ability of PBMC to respond to stimuli (selected for their ability to optimally induce each cytokine) in terms of IL-2, IL-4 and IL-10 production in patients with both high and low CD4 cell counts; and (iii) conversely, HIV infection led to an overproduction of IFN-gamma in patients with high CD4 counts; patients with low CD4 produced normal levels of IFN-gamma. CONCLUSIONS: Our observations did not suggest polarization of the type 1/type 2 cytokine profile in HIV patients. Instead, the data suggested more complex changes to type 1/type 2 cytokine patterns in HIV infection than originally proposed by the TH1/TH2 dichotomy. PMID- 9342067 TI - Effect of recombinant human granulocyte-macrophage colony-stimulating factor on HIV-related leukopenia: a randomized, controlled clinical study. AB - OBJECTIVE: To assess the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on white blood cell (WBC) count and on the rate of opportunistic infections in a large and selected population of leukopenic HIV-positive patients compared with non-treated controls. DESIGN: Open-label, randomized, comparative clinical study. SETTING: University hospitals and AIDS centres. PATIENTS AND METHODS: One hundred and twenty-three leukopenic HIV-positive patients received recombinant human GM-CSF (300 microg subcutaneously daily for 1 week, and 150 microg subcutaneously two times weekly for 11 weeks thereafter); the control group comprised 121 non-treated leukopenic HIV-positive patients. A complete blood cell count with differential, platelet count, reticulocyte count, and CD4+ and CD8+ T-cell subset counts were performed in both patient groups at baseline and at weeks 1, 12 and 24. RESULTS: The administration of GM-CSF resulted in a significant increase of WBC count in patients compared with non-treated controls. Total leukocyte count increased by 22% at week 1 and by 65% at week 12 compared with baseline levels; a 20% increase of total leukocyte count was still present at week 24. Increases of neutrophils, eosinophils and monocytes were responsible for the majority of the increase in WBC count. Opportunistic infections occurred in 61.7% of GM-CSF-treated patients and in 72% of the patients of the control group (relative risk, 0.86; 95% confidence interval, 0.72-1.03; P = 0.123). Mild flu-like side-effects were observed in most patients receiving GM-CSF, although they were not sufficiently severe to warrant withdrawal from the study. CONCLUSIONS: GM-CSF was well tolerated and biologically active in leukopenic HIV positive patients, with a significant, although time-limited, increase of WBC count compared with non-treated patients. The administration of this growth factor should be considered in ameliorating the myelosuppression observed with some cell-cycle-specific antiviral and anti-neoplastic agents. PMID- 9342068 TI - Liposomal amphotericin B (AmBisome) compared with amphotericin B both followed by oral fluconazole in the treatment of AIDS-associated cryptococcal meningitis. AB - OBJECTIVE: Amphotericin B deoxycholate initial therapy and fluconazole maintenance therapy is the treatment of choice for AIDS-associated cryptococcal meningitis. However, the administration of amphotericin B is associated with considerable toxicity. A potential strategy for reducing the toxicity and increasing the therapeutic index of amphotericin B is the use of lipid formulations of this drug. DESIGN AND METHODS: HIV-infected patients with cryptococcal meningitis were randomized to treatment with either liposomal amphotericin B (AmBisome) 4 mg/kg daily or standard amphotericin B 0.7 mg/kg daily for 3 weeks, each followed by fluconazole 400 mg daily for 7 weeks. During the first 3 weeks, clinical efficacy was assessed daily. Mycological response was primarily evaluated by cerebrospinal fluid (CSF) cultures at days 7, 14, 21 and 70. RESULTS: Of the 28 evaluable patients, 15 were assigned to receive AmBisome and 13 to receive amphotericin B. Baseline characteristics were comparable. The time to and the rate of clinical response were the same in both arms. AmBisome therapy resulted in a CSF culture conversion within 7 days in six out of 15 patients versus one out of 12 amphotericin B-treated patients (P = 0.09), within 14 days in 10 out of 15 AmBisome patients versus one out of nine amphotericin B patients (P = 0.01), and within 21 days in 11 out of 15 AmBisome patients versus three out of eight amphotericin B patients (P = 0.19). When Kaplan-Meier estimates were used to compare time to CSF culture conversion, AmBisome was more effective (P < 0.05; median time between 7 and 14 days for AmBisome versus > 21 days for amphotericin B). AmBisome was significantly less nephrotoxic. CONCLUSIONS: A 3-week course of 4 mg/kg AmBisome resulted in a significantly earlier CSF culture conversion than 0.7 mg/kg amphotericin B, had equal clinical efficacy and was significantly less nephrotoxic when used for the treatment of primary episodes of AIDS-associated cryptococcal meningitis. PMID- 9342070 TI - Increased losses of CD4+CD45RA+ cells in late stages of HIV infection is related to increased risk of death: evidence from a cohort of 347 HIV-infected individuals. AB - OBJECTIVE: To examine changes in the distribution of CD4+CD45RA+ (naive) and CD4+CD45RO+ (memory) lymphocytes in various stages of HIV infection and the effect of these changes on disease progression. DESIGN AND METHODS: Expression of CD45RA+ and CD45RO+ on CD4+ lymphocytes was analysed by flow cytometry in a prospectively followed cohort of 300 HIV-infected individuals (median follow-up time, 2.90 years; range, 0.02-4.54 years) and in a group of 102 age- and sex matched uninfected controls. Survival analysis was performed considering AIDS development and death as endpoints. RESULTS: The median CD4+CD45RA+/CD45RO+ ratio was 1.3 (25-75% quartiles, 0.9-2.4) in controls; it was increased to 1.8 (1.1 2.5) in 40 HIV-infected individuals with CD4+ cell counts > 500 x 10(6)/l (P < 0.05); it was similar at 1.4 (0.8-2.0) in 106 HIV-infected individuals with CD4+ cell counts of 200-500 x 10(6)/l; and it was decreased to 0.9 (0.5-1.4) in 154 HIV-infected individuals with CD4+ cell counts < 200 x 10(6)/l (P < 10[-6]). When fitted in a Cox model adjusting for the total number of CD4+ cells and age a lower concentration of CD4+CD45RA+ cells was associated with an increased risk of dying. The concentration of CD4+CD45RO+ cells was not significantly associated with AIDS or death in age- and CD4+ cell count-adjusted Cox models. CONCLUSIONS: This study confirms a selective loss of memory CD4+ cells early in HIV infection followed by increased loss of naive CD4+ cells in later stages of the infection. The loss of naive CD4+ cells seems to be important in the pathogenesis of terminal HIV infection. PMID- 9342069 TI - Susceptibility to levofloxacin of Myocobacterium tuberculosis isolates from patients with HIV-related tuberculosis and characterization of a strain with levofloxacin monoresistance. Community Programs for Clinical Research on AIDS 019 and the AIDS Clinical Trials Group 222 Protocol Team. AB - OBJECTIVE: To characterize the susceptibility to levofloxacin of clinical isolates of Mycobacterium tuberculosis (MTB) obtained from patients with HIV related tuberculosis and to characterize the molecular genetics of levofloxacin resistance. DESIGN AND METHODS: Isolates from culture-positive patients in a United States multicenter trial of HIV-related TB were tested for susceptibility to levofloxacin by minimum inhibitory concentration (MIC) determinations in Bactec 7H12 broth. Automated sequencing of the resistance determining region of gyrA was performed. RESULTS: Of the 135 baseline MTB isolates tested, 134 (99%; 95% exact binomial confidence interval, 95.9-99.9%) were susceptible to levofloxacin with an MIC < or = 1.0 microg/ml. We identified a previously unrecognized mis-sense mutation occurring at codon 88 of gyrA in a levofloxacin mono-resistant MTB isolate obtained from a patient with AIDS who had received ofloxacin for 8 months prior to the diagnosis of tuberculosis. CONCLUSIONS: Clinical MTB isolates from HIV-infected patients were generally susceptible to levofloxacin. However, the identification of a clinical isolate with mono resistance to levofloxacin highlights the need for circumspection in the use of fluoroquinolones in the setting of potential HIV-related tuberculosis and for monitoring of rates of resistance of MTB isolates to fluoroquinolones. PMID- 9342073 TI - Increasing diversity of HIV-1M serotypes in French blood donors over a 10-year period (1985-1995). Retrovirus Study Group of the French Society of Blood Transfusion. AB - OBJECTIVE: Phylogenetic analysis of gene sequences of HIV-1 has led to the classification of isolates into a major group (M) of viruses, itself divided into subtypes (A to I), and a minor group (O) of rare isolates. Subtype B viruses are the most prevalent in Western countries but little is known about the dynamics of diffusion of the other subtypes in these regions. The prevalence of B subtypes and non-B subtypes in French blood donors between 1985 and 1995 was evaluated. METHODS: A retrospective study was conducted in 490 blood donors, identified as positive for antibody to HIV-1, by twelve French blood banks between 1985 and 1995. Serological subtyping was performed with a subtype-specific enzyme immunoassay, the reliability for genotyping of which has been demonstrated previously. RESULTS: Of 450 typable samples, 48 (10.7%) were non-B subtypes. Non B reactive samples were found in all of the regions. An increasing prevalence of individuals infected by non-B viruses was observed, from approximately 4% in the early period to more than 20% in 1994-1995 (P = 0.0004). Non-B viruses did not appear to be restricted to patients with direct or indirect epidemiological links to non-European populations. CONCLUSION: We observed an increasing diversity of HIV-1 strains in the population of blood donors residing in France. This stresses the necessity to broaden the surveillance of HIV-1 diversity in order to improve measures to prevent HIV-1 infections. PMID- 9342071 TI - Longitudinal evaluation of severely anemic children in Kenya: the effect of transfusion on mortality and hematologic recovery. AB - OBJECTIVE: To determine the effect of transfusion on hematologic recovery and mortality among severely anemic children during and after hospitalization in rural Kenya. DESIGN: Prospective cohort. METHODS: We collected clinical and laboratory information on all severely anemic children (hemoglobin < 5.0 g/dl) and a 33% sample of children with hemoglobin < or = 5.0 g/dl who were admitted to the pediatric ward of a rural Kenyan hospital during a 6 month study period. Children were followed during hospitalization and at 4 and 8 weeks after admission. RESULTS: Overall, 303 (25%) of the 1223 hospitalized children had hemoglobin < 5.0 g/dl, 30% of whom died during the study period. Severely anemic children who were transfused had a higher mean hemoglobin level at discharge (9.0 g/dl) than non-transfused children (5.8 g/dl, P < 0.001) and maintained a higher mean hemoglobin during the 8-week follow-up period. However, the presence of malaria parasitemia on follow-up negated the benefit of transfusion on hematologic recovery at both 4- and 8-week visits (longitudinal linear model, least square means, P > 0.05). Transfusion was associated with improved survival among children with respiratory distress who received transfusions within the first 2 days of hospitalization. CONCLUSIONS: The use of transfusion can be improved by targeting use of blood to severely anemic children with cardiorespiratory compromise, improving immediate availability of blood, and treating severely anemic children with effective antimalarial therapy. PMID- 9342074 TI - CD4 surveillance in Scotland: perspectives on severe HIV-related immunodeficiency. CD4 Collaborative Group. AB - OBJECTIVE: To analyse factors that influence the following: the square root of first CD4 cell count; which individuals are severely immunodeficient at or before the start of monitoring; progression from the date of the earlier of two consecutive CD4 counts < or = 200 x 10(6)/l (termed as CD200) to AIDS. SETTING: Scotland's HIV Immunology Laboratories and the Scottish Centre for Infection and Environmental Health. PATIENTS: A total of 1679 adult HIV patients in Scotland to 31 December 1994 who had ever had a CD4 cell count < or = 500 x 10(6)/l or who had developed AIDS without any immunological monitoring, of whom 912 had developed severe HIV-related immunodeficiency (i.e., were CD200/AIDS cases). RESULTS: Square-root first CD4 count was higher in women (by 2.1; SE, 0.5), in injecting drug users (IDU; by 1.8; SE, 0.5) and in younger patients (by 1.5 per 10 years; SE, 0.2), but reduced by 1.5 (SE, 0.1) per calendar year of recruitment, although it was relatively higher (by 3.8; SE, 0.8) for Edinburgh patients recruited in 1993-1994: at least 30% (nine out of 28) of new Edinburgh City Hospital patients in 1993-1994 with a first CD4 count of > or = 500 x 10(6)/l had seroconverted within the past 5 years. Two-thirds of non-IDU (67%; 348 out of 517) were already severely immunodeficient at or before the start of immunological monitoring, in contrast with only 31% of IDU CD200/AIDS cases. Overall, the odds on the CD200 date also being the date of first CD4 count have increased in recent times [log(e)(odds per calendar year of CD200 diagnosis), 0.14; SE, 0.05]. Analysis excluding patients whose AIDS diagnosis or follow-up time was within 1 month of the CD200/AIDS date supported a modest prolongation of the CD200/AIDS to AIDS interval for patients diagnosed with severe HIV-related immunodeficiency in the period 1989-1991 (log(e)[relative risk (RR)], -0.46; SE, 0.22). Similarly, year effects were evident on progression from CD500 to CD200/AIDS [log(e)(RR), -0.55; SE, 0.17) for CD500 cases diagnosed in 1989, and these year effects doubled in 1990-1992. CONCLUSIONS: Minimal CD4 data were hypothesis-generating about region, risk group and calendar year. Lower bound for recent HIV incidence can be derived from new patients with first CD4 cell count above 500 x 10(6)/l if seroconversion intervals are available for a proportion. PMID- 9342072 TI - Actual versus perceived HIV status, sexual behaviors and predictors of unprotected sex among young gay and bisexual men who identify as HIV-negative, HIV-positive and untested. AB - OBJECTIVES: To compare the prevalence and predictors of HIV sexual risk behavior among young gay and bisexual men who perceived themselves to be HIV-negative, HIV positive, or who were untested. DESIGN: Population-based sample of young gay and bisexual men. METHODS: Using multi-stage probability sampling, 408 gay and bisexual men aged 18-29 years in San Francisco were recruited and interviewed, and blood samples for HIV-testing from 364 participants were obtained. RESULTS: HIV prevalence was 18.7%, although 25% of the men who were HIV-positive did not know it. Thirty-seven per cent reported engaging in unprotected anal intercourse during the past year, including 59% of the men who knew they were HIV-positive, 35% of the men who perceived themselves HIV-negative and 28% of the untested men. Logistic regressions found similar predictors of unprotected intercourse for HIV negatives and HIV-positives, including sexual impulsivity, substance use, sexual enjoyment and communication problems. CONCLUSIONS: The high rates of unprotected intercourse, particularly among the HIV-positive men, attest to the urgent need for HIV-prevention interventions for young gay and bisexual men. The findings suggest that many of the important variables to target in interventions are similar for both HIV-positive and HIV-negative men. PMID- 9342075 TI - Knowledge of HIV transmission and sexual behavior of college students in Pune, India. PMID- 9342076 TI - Nimodipine plus zidovudine versus zidovudine alone in the treatment of HIV-1 associated cognitive deficits. PMID- 9342077 TI - Evaluation of contrast-enhancing brain lesions in AIDS patients by means of Epstein-Barr virus detection in cerebrospinal fluid and 201thallium single photon emission tomography. PMID- 9342078 TI - Stavudine, lamivudine and indinavir in children with advanced HIV-1 infection: preliminary experience. PMID- 9342080 TI - Regression of progressive multifocal encephalopathy with highly active antiretroviral therapy. PMID- 9342079 TI - Nonoxynol-9 as a xenobiotic with endocrine activity. PMID- 9342081 TI - Drug resistance mutations as predictors of phenotypic zidovudine resistance in HIV-1 infection. PMID- 9342082 TI - Diagnosis of toxoplasmic encephalitis in HIV-infected patients. PMID- 9342083 TI - Human herpesvirus DNA in prostate and semen from HIV-negative individuals in Italy. PMID- 9342084 TI - Large proportion of non-B HIV-1 subtypes and presence of zidovudine resistance mutations among German seroconvertors. PMID- 9342085 TI - Prevalence and distribution of foot lesions in finishing pigs in south-west England. AB - This paper gives the first estimate of the prevalence and distribution of foot lesions in finishing pigs in the south-west of England for 33 years. It was based on the examination of 4038 finishing pigs from 21 units. There were 3727 out of 3974 (93.8 per cent) pigs with at least one foot lesion. The prevalence of different lesions was: toe erosion (33.0 per cent), sole erosion (62.1 per cent), heel erosion (13.0 per cent), heel flaps (14.4 per cent), white line lesions (55.4 per cent), false sand cracks (23.9 per cent) and wall separation (11.5 per cent). The hind feet were more commonly affected than the front feet, and on each foot the lateral digits were significantly more frequently affected than the medial digits. Sole erosions, heel flaps, wall separation and false sand cracks were more frequently observed on the lateral than the medial digit. Digits with sole erosions were significantly more likely to have heel flaps, white line lesions or false sand cracks than those without sole erosions. Furthermore, there was a within digit association between overgrown hooves and toe erosion. The effects of weightbearing, gait and environment on the development of foot lesions is discussed. PMID- 9342086 TI - Prophylactic efficacy of persistent anthelmintics against challenge with drug resistant and susceptible Ostertagia circumcincta. AB - Three groups of newly-weaned Romney lambs were given either a standard oral dose of albendazole, a controlled-release capsule containing albendazole, or a standard oral dose of moxidectin. At 10, 20, 30 and 40 days after treatment, sub groups of lambs were given 10,000 infective-stage larvae of either a drug resistant or a drug-susceptible strain of Ostertagia circumcincta. The recommended oral dose of albendazole removed 32 per cent of the resistant strain and over 99.9 per cent of the susceptible O. circumcincta. The recommended oral dose of moxidectin removed 91 per cent of the resistant strain and over 99.9 per cent of the susceptible parasites. None of the lambs treated with controlled release capsules was challenged at 20 or 30 days after treatment. Twenty-one days after challenge, samples of faeces were taken to determine the presence of nematode eggs and cultured to establish the proportion of eggs developing to infective-stage larvae (L3). Abomasa were recovered after slaughter and worm burdens determined. In the lambs given controlled-release capsules only the resistant parasites were able to establish, and there were significantly fewer than in the lambs treated orally with albendazole. The proportion of the eggs from resistant parasites which developed to L3 was not reduced by the presence of the capsules. Oral moxidectin provided no protection against the establishment of the resistant strain and viable L3 were recovered after challenge with resistant parasites 10 days after treatment; however, the establishment of susceptible O. circumcincta was reduced by more than 99 per cent. The establishment of the susceptible parasites in the lambs treated with moxidectin increased with time and was not significantly lower than in the other groups by 30 days after treatment. PMID- 9342087 TI - A review of viruses affecting raptors. AB - Outbreaks of viral diseases have been diagnosed more commonly in raptors in recent years. The practice of feeding carnivorous birds with food derived from other birds exposes them directly to a wide range of potential pathogens. Some viruses which are avirulent in their natural host are known to be more pathogenic when they cross the species barrier. Compromised immunity due to stress or inbreeding may further increase the disease risk. Traditional feeding methods may need to be re-appraised and changed in view of this risk. This paper reviews the literature on viral diseases of raptors and provides additional clinicopathological observations from unpublished cases. PMID- 9342088 TI - Non-market costs of rabies policy. PMID- 9342089 TI - Extensive pleural effusion associated with diffuse fibrosing alveolitis in an aged beef cow. PMID- 9342090 TI - Bovine respiratory syncytial virus infection in lactating cows. PMID- 9342091 TI - Rabies and quarantine. PMID- 9342092 TI - Uterus unicornis in mares. PMID- 9342093 TI - Qualifications in companion animal behaviour. PMID- 9342094 TI - Mare recovers from further injury after tear repair. PMID- 9342095 TI - Nuclear medicine under inspection. PMID- 9342096 TI - Non-invasive scintigraphic monitoring of gene expression in a HSV-1 thymidine kinase gene therapy model. AB - In vivo transduction of the herpes simplex virus type-1 thymidine kinase (HSV-1 TK) gene and subsequent administration of antiviral drugs such as ganciclovir has emerged as a promising gene therapy protocol for proliferative disorders. However, the evaluation of HSV-1 TK gene expression in transduced tissue has relied on invasive techniques for detection. We now report that HSV-1 TK expression can be detected non-invasively using scintigraphy. The radioiodinated nucleoside analogue, (E)-5-(2-iodovinyl)-2'-fluoro-2'-deoxyuridine (IVFRU), becomes metabolically trapped in tumour cells transduced with the HSV-1 TK gene on a retroviral vector. Selective phosphorylation of radiolabelled IVFRU by HSV-1 TK results in elevated radioactivity in HSV-1 TK-expressing cells in vitro and in vivo relative to cells lacking the HSV-1 TK gene. Due to low non-target tissue uptake, unambiguous imaging of HSV-1 TK-expressing tumours in mice is possible with labelled IVFRU. We have monitored the process of tumour regression non invasively during ganciclovir treatment using labelled IVFRU and scintigraphy. PMID- 9342097 TI - Oestrogen receptors in meningiomas: a correlative PET and immunohistochemical study. AB - There have been a number of indications that sex hormones can affect the rate of growth of meningiomas during pregnancy. The presence of oestrogen or progesterone receptors in meningiomas and the influence of sex hormones upon cell cultures derived from human meningiomas have been extensively investigated. The results have been controversial, with most of the discussion centring upon the presence and possible role of oestrogen receptors. The aim of the present study was to assess oestrogen receptors in human meningiomas with 16alpha[l8F]fluoro-17beta oestradiol ([18F]FES) and positron emission tomography (PET). With this purpose in mind, we measured the regional brain uptake of [18F]FES in six patients with a neuroradiological and histological diagnosis of meningioma, comparing the in vivo PET data with the immunohistological analysis of oestrogen receptors performed on formalin-fixed, paraffin-embedded tissue obtained at surgery. Two analyses were used for the in vivo measurement of [18F]FES binding to oestrogen receptors: the ratio of tumour activity to that of normal tissue (T/NT), calculated 90 min after tracer injection, and the ratio between the equilibrium distribution volume (DV) in normal and pathological tissues, calculated by means of a graphical kinetic analysis. PET studies demonstrated a marked uptake of [18F]FES by the tumour in four of the six patients. Immunohistochemical assay using a manual staining method capable of detecting oestrogen receptors at a level of > 10 pmol mg(-1) of protein detected only sparse immunostaining in one of the six meningiomas. Distinct albeit weak immunostaining was demonstrated in five of the six meningiomas when the sensitivity of the immunohistochemical assay was increased to < 10 pmol mg(-1) of protein by use of an automated staining method (Bioteck 1000). Comparison of the in vivo and immunohistochemical results showed a correlation in five of the six patients, thus indicating the high sensitivity of [18F]FES for the in vivo evaluation of oestrogen receptor expression. PMID- 9342098 TI - A comparison of 111In-octreotide and 67Ga scintigraphy in malignant lymphoma. AB - We compared 111In-octreotide and 67Ga scintigraphy for staging malignant lymphoma. In 11 patients, planar imaging was performed 4 and 24 h after the injection of 111In-octreotide and 48 and 72 h after the administration of 67Ga. Radiological and clinical data were used as the 'gold standard', resulting in the identification of 26 lesions. Twenty-three (88%) of these lesions were detected by 67Ga and 18 (69%) by 111In scintigraphy. All 14 supra-diaphragmatic lesions were detected by 67Ga and 13 by 111In. In the intra-abdominal areas, only two of eight known localizations were identified by 111In scintigraphy, whereas 67Ga uptake was seen in six of them. In the inguinal regions, both tracers detected three of four lesions. Of the eight lesions of low-grade malignancy, seven were visualized by 67Ga and five by 111In-octreotide imaging. For the intermediate and high-grade lymphomas, 67Ga showed a similar detection rate (16/18 lesions), whereas 111In was only able to detect 13 lesions. We conclude that 111In octreotide provides less information than 67Ga scintigraphy and conventional staging modalities. The best results for both tracers were observed above the diaphragm. The 67Ga results were more reliable for the detection of infra diaphragmatic lesions. PMID- 9342099 TI - Evaluation of complications of kidney transplantation using artificial neural networks. AB - The aim of this study was to develop an artificial neural network (ANN) to differentiate between rejection, acute tubular necrosis (ATN) and normally functioning kidneys in a group of patients with renal transplants. The performance of ANN was compared with that of an experienced observer using a database of 35 patients' records, each of which included 12 quantitative parameters derived from renograms and clinical data as well as a clinical evaluation. These findings were encoded as features for a three-layered neural network to predict the outcome of biopsy or clinical diagnosis. The network was trained and tested using the jackknife method and its performance was then compared to that of a radiologist. The network was able to correctly classify 31 of the 35 original cases and gave a better diagnostic accuracy (88%) than the radiologist (83%), by showing an association between the quantitative data and the corresponding pathological results (r = 0.78, P < 0.001). We conclude that an ANN can be trained to differentiate rejection from acute tubular necrosis, as well as normally functioning transplants, with a reasonable degree of accuracy. PMID- 9342100 TI - A review of captopril renal scintigraphy and its effect on patient management. AB - Renal artery stenosis is an important and potentially curable cause of hypertension. Captopril renography is now recognized to have a high sensitivity and specificity in its diagnosis. Ultimately, however, the result is of little benefit if it does not lead to a change in patient management. To assess how patient management was changed following the result of a captopril renogram, we reviewed the notes of 95 patients who had undergone this test over a 5 year period to identify renal artery stenosis. Of these patients, significant renal artery stenosis was suggested in 16 (17%), of whom only 9 (56%) underwent a change in management (7 proceeding to angiography with or without angioplasty, 2 having alterations in medication). In the 67 patients who had a negative renogram, 16 (24%) had an alteration in management (13 angiography, 3 altered drug treatment). Finally, of the 12 patients who had a non-diagnostic renogram, 7 (60%) had a management change (3 angiography, 4 altered drug therapy). Our results suggest that, despite evidence from the literature that captopril renography is both sensitive and specific for renal artery stenosis, clinicians still rely on other factors when determining who has significant stenosis and, therefore, who should proceed to a further investigation or have a change in medication. Ultimately, this reduces the clinical value of the test at present. PMID- 9342101 TI - Estimation of glomerular filtration rate with 99Tc(m)-DTPA: a comparative assessment of simplified methods. AB - In 40 adult patients undergoing gamma camera renography, glomerular filtration rate (GFR) was measured using simplified 99Tc(m)-DTPA methods (i.e. a personal modification of the 'slope' method which does not require dose calibration, Gates' method and Carlsen's method) and compared to reference results (obtained using Sapirstein's formula and Russell's two-sample method with 51Cr-EDTA). Estimation of GFR from plasma creatinine (the Cockroft-Gault formula) was also carried out. Bias and imprecision of the simplified estimates were determined by the Bland-Altman method. The GFR values of the 'slope' method correlated best with the reference values (R2 = 0.88, S.E.E. = 11.3 ml min[-1]). Correlation of the two methods based on external determination with the gamma camera was no better at estimating GFR than that from plasma creatinine. Moreover, Gates' method underestimated GFR at all levels between 25 and 150 ml min(-1), while Carlsen's method overestimated at low levels and underestimated at high levels. The bias was as follows (ml): Cockroft-Gault 2.4; 'slope' -4.1; Carlsen 7.5; Gates 16.7. The imprecision was as follows (ml): 'slope' 11.8; Cockroft-Gault 16.4; Carlsen 20.5; Gates 22.8. We conclude that our modification of the slope method correlated best with the reference results, and would appear suitable for routine practice because of the small error involved. When performing sequential renal scintigraphy, it can also be used for a quick check of dubious data based on gamma camera methods. PMID- 9342102 TI - Quantification of the atrial contribution to diastolic filling during radionuclide angiography. AB - To define exactly the onset of late diastolic filling with respect to atrial contraction, the atrial contribution (AC) to left ventricular filling was quantified in 34 patients with a variety of diseases using radionuclide angiography. From the time-activity curve and its first derivative, a flow-volume loop was constructed. Using the flow-volume loop, the period between minimal flow and the moment of maximal end-diastolic counts was defined as the AC-interval and correlated with the PQ-interval on the electrocardiogram. The relative filling volumes within these time periods were closely related in all patients (r = 0.99, P < 0.0001). The correlation between the PQ-interval and AC-interval was also statistically significant (r = 0.82, P < 0.0001). In a subset of patients, the PQ interval and AC-interval were not exactly the same. In these patients, the AC interval was always longer than the PQ-interval, indicating the existence of passive diastasis flow before the onset of atrial contraction. This was more apparent in patients with low heart rates than in those with high heart rates. Despite the close correlation between the PQ-interval on the electrocardiogram and the AC-interval of the flow-volume loop, they may represent different entities. We conclude that the PQ-interval is better than the AC-interval for determining the moment of onset of atrial activity during radionuclide angiography. PMID- 9342103 TI - Simulation of the stripe sign in a scintigraphic model of the lungs. AB - Using a virtual model of the lungs, we investigated the nature of the 'stripe sign' which is sometimes encountered in pulmonary scintigraphy. A model of the segmental anatomy of the lungs was developed from a number of sources and counts generated within the phantom by Monte-Carlo simulation of photon emission. Multiple segmental and subsegmental defects were created in both lungs and submitted for blinded reporting of the 'stripe sign'. Images were resubmitted for reporting with the contralateral lung removed. The stripe sign was reported in 32 of the 117 studies performed. Nearly half of these were present in defects involving approximately 25% of a segment and the sign was most commonly seen in the lateral projection. Removal of activity from the contralateral lung abolished the sign in only 2 of 32 cases. We conclude that shine through of activity from the contralateral lung is a mechanism rarely responsible for the stripe sign. Most occurrences of the sign are due to interposition of activity from unaffected areas of the same lung between the defect and the periphery of the lung. Orientation of the segments, particularly in the lung bases, accounts for the lateral projection being the most common view in which the sign is present. PMID- 9342104 TI - Selection of collimator for rCBF studies and evaluation of triple-headed SPET using noise-resolution plots. AB - We investigated the effect of collimator selection on image quality in regional cerebral blood flow (rCBF) studies of the brain performed with 99Tc(m)-HMPAO. A triple-headed SPET system (GE/CGR Neurocam) was used, together with three sets of parallel-hole collimators - general-purpose (GP), high-resolution (HR) and ultra high-resolution (UHR). Two image quality parameters were used to describe the image quality, namely, noise and resolution. Noise was measured in experimental and Monte-Carlo simulated SPET studies of a cylinder phantom of uniform activity as the pixel root mean square error (RMS) and as the coefficient of variation (CV) of quantitative rCBF values. Resolution was measured as full-width at half maximum in experimental SPET studies of a line-source. Plots of noise versus resolution for the different collimators were obtained by varying the cut-off frequency of the Hanning filter applied in the reconstruction of transaxial slices. From these noise-resolution plots, we were able to determine which collimator gave the best resolution for a specific noise level. A lowest reasonable noise level may be established by comparison with the inter-observer CV of the quantification method. PMID- 9342105 TI - A study of dementia in adults with Down's syndrome using 99Tc(m)-HMPAO SPET. AB - Twenty-six people with Down's syndrome (DS) were investigated using 99Tc(m) hexamethylpropylene amine oxime (99Tc(m)-HMPAO) and single photon emission tomography (SPET). Dementia was diagnosed using a structured carer interview giving a deterioration score and by studying the case notes. Five subjects were clinically demented, 7 showed mild deterioration and 14 no deterioration. Increased deterioration correlated with advancing age (correlation coefficient 0.5425; P<0.02), but there was no significant difference between older (>40 years) and younger (<40 years) patients. Only one of the subjects with dementia had a regional cerebral blood flow (rCBF) abnormality that was of the dementia of Alzheimer type. Three subjects with mild deterioration and three with no deterioration had abnormal SPET scans. There was no association between the SPET abnormality and clinical dementia or with evidence of deterioration. PMID- 9342106 TI - A UK survey of nuclear medicine imaging performance using the TransBone anthropomorphic phantom. AB - In March 1993, the British Nuclear Medicine Society was approached and agreed to take part in an international nuclear medicine quality survey organized jointly by the World Health Organization and the International Atomic Energy Agency. The study involved the use of a novel anthropomorphic transmission phantom, which contained a number of abnormalities of varying contrast. The UK part of the survey was undertaken during April and May 1994. A total of 14 departments took part in the study, using a total of 25 gamma cameras. For each gamma camera tested, imaging was performed using the same camera set-up, imaging and hard copy parameters normally used for recording bone images. The test image was then reported by the usual clinical image reporter according to a fixed protocol and a 4-point scoring system. The results varied widely, with 62% of all reportable locations using all four possible scores between the centres being tested. The sensitivity and specificity ranged from 40 to 80% and 59 to 100% respectively. Calculated ROC areas ranged from 65 to 94% with a mean of 80%. The survey methodology means that a relatively large number of factors affect the overall results, of which the main factor appears to be reporting accuracy. PMID- 9342107 TI - Application of a gamma extremity monitoring system in a radiopharmaceutical dispensary. AB - A new gamma extremity monitoring system (GEMS) was used to assess finger doses of staff working in a hospital radionuclide dispensary. The system is designed to give a continuous readout of dose rate from a small probe which may be attached to a finger. It allows the contributions to radiation dose to the fingers from different parts of a procedure to be measured for the first time and the detailed pattern of radiation exposure to be determined. The dose reduction benefits of small changes in procedure and the use of syringe shields were easily demonstrated after monitoring staff for a few sessions using GEMS, which would not have been possible using thermoluminescent or other integrating dosemeters. GEMS was calibrated against 99Tc(m), 241Am, 137Cs and 131I. The main disadvantage of the system is that the response of the detector increases significantly at lower radiation energies, with that at 60 keV being approximately 70 times the response at 662 keV. In addition, the response of the current detector is nonlinear and saturates around 7000 counts s(-1), which corresponds to a dose rate of 2200 microGy h(-1) for 99Tc(m). Despite these drawbacks, GEMS can play a significant part in the analysis of finger dose patterns and assist in dose reduction. PMID- 9342108 TI - A risk assessment of multiple use of an aerosol system compared with single use for lung ventilation scans. AB - Over 300 99Tc(m)-DTPA aerosol lung ventilation scans are performed at Leicester Royal Infirmary each year. Current practice is to re-use the nebulizer circuits up to five times over a maximum of 1 week. Following a visit from the hospital infection control team, this practice was questioned and, therefore, a risk assessment was carried out. This compared the risks of multiple use of ventilation circuits with single use. A survey was also carried out to try and establish current practice in other departments. The risk assesment showed that both single and multiple use of nebulizer circuits can have a high risk. The risks associated with the former were radiation safety and/or financial, and the risk with the latter was cross-infection. The survey showed that multiple use is certainly common practice within the UK. Therefore, in coming to a decision on which practice to adopt, these risks must be considered most carefully in the context of the local environment. PMID- 9342116 TI - Spectroscopic and metabolic effects of MnCl2 and MnDPDP on the isolated and perfused rat heart. AB - RATIONALE AND OBJECTIVES: Several works have shown that the hepatobiliary magnetic resonance imaging contrast agent manganese dipyridoxyl diphosphate (MnDPDP) partly releases its metallic ion and exhibits cardiovascular effects that are supposed to arise from the free manganese ions (Mn++). In the current study, the cellular internalization of Mn by the isolated rat heart is monitored through the mechanical function of the organ and the relative broadening of the P 31 nuclear magnetic resonances. METHODS: Rat hearts were perfused with manganese chloride (MnCl2; 15 and 25 microM) or MnDPDP (25 microM). Variations of the linewidths, heights, and surfaces of phosphocreatine and adenosine triphosphate peaks were monitored. Cardiac function was monitored simultaneously through heart rate, left ventricular pressure, and coronary flow. RESULTS: Influx of Mn++ induces a significant broadening of the P-31 resonances of adenosine triphosphate and phosphocreatine because of a strong scalar paramagnetic interaction between the nuclei and the ion. Compared with MnDPDP administered at the same concentration, MnCl2 induced a more pronounced and dose-dependent line broadening as well as a coronary vasodilation. Calcium channel blockers (nifedipine and verapamil) and EDTA inhibit MnCl2 influx. Similarly, verapamil, EDTA, and DPDP reduce the alterations provoked by MnDPDP. CONCLUSIONS: The effects of MnDPDP are smaller but of the same type than those induced by MnCl2. Their inhibition by calcium channel blockers (verapamil and nifedipine) and by an excess of strong chelators such as DPDP or EDTA confirms that they originate from a partial release of Mn++ by the contrast agent. PMID- 9342117 TI - Contrast-enhanced magnetic resonance imaging in a spinal epidural tumor model. AB - RATIONALE AND OBJECTIVES: A spinal epidural tumor model was developed, using the VX-2 adenocarcinoma in rabbits, to assess the strengths and weaknesses of magnetic resonance (MR) as a cross-sectional imaging modality for the evaluation of epidural neoplastic disease. High-resolution MR images were acquired both before and after intravenous gadolinium chelate injection, assessing lesion detectability and efficacy of imaging technique. METHODS: An adenocarcinoma tumor (VX-2) was produced in the epidural space of six New Zealand White rabbits and subsequently studied on a 1.5 tesla whole body MR scanner. VX-2 tumor tissue was removed from the thigh of a carrier rabbit, minced, and screened. Under fluoroscopic guidance, 0.2 mL of the tumor preparation was then injected into the epidural space of the experimental rabbits. The injection was performed at the L5 6 level using an epidural needle and polyethylene tubing sleeved within the needle. The rabbits were imaged using a circular small parts surface coil 5 to 15 days after the epidural injection. In all six animals, one complete MR exam was obtained within the time frame of days 9 to 11. T1- and T2-weighted axial scans were obtained before contrast injection, with the T1 scans acquired both with and without fat saturation. Postcontrast T1 scans also were obtained, using fat saturation, after the injection of 0.1 and 0.3 (cumulative dose) mmol/kg gadoteridol (Gd HP-DO3A; ProHance) in all animals. The film images were interpreted in a prospective fashion by a single neuroradiologist who was masked to imaging technique and contrast dosing. The digital data was analyzed by region of interest measurement. At the end of the imaging studies, the animals were sacrificed and the epidural lesion confirmed by gross and microscopic exam. RESULTS: On a prospective masked read of the MR films, epidural tumor was depicted best on postcontrast fat saturation T1-weighted scans using a cumulative contrast dose of 0.3 mmol/kg. Substantial contrast enhancement of the tumor was observed in all instances on postcontrast scans. The precontrast T1-weighted scan was least efficacious for lesion identification and differentiation from the compressed spinal cord. Depending on the pulse sequence used, one (T2-weighted) to three (T1-weighted without fat saturation) of the lesions could not be identified prospectively on precontrast scans. Lesion growth with time after implantation was chronicled by MR imaging, accompanied by progression of symptoms. On region of interest analysis, differentiation of epidural tumor from normal cord was greatest (11.6 +/- 6.1) on postcontrast scans using a cumulative contrast dose of 0.3 mmol/kg. The level of differentiation achieved was twice that of postcontrast scans using a contrast dose of 0.1 mmol/kg (5.9 +/- 3.6). These results were superior on statistical analysis to that with all other scan techniques (P = 0.002-0.0005). Cord and tumor could not be differentiated on the basis of signal intensity, with any statistical significance, using precontrast T1 and T2 scans. The lesions were confirmed in each animal by gross and microscopic exam. On inspection of the gross specimen, the tumors were noted to be located in the epidural space and to cause cord compression. On microscopic exam, the tumor was composed of epithelial cells that were moderately pleomorphic. CONCLUSIONS: In the New Zealand White rabbit, an epidural tumor could be created consistently using the described percutaneous approach. These lesions are suitable for MR imaging studies, examining lesion detectability and efficacy of imaging technique. The lesions created in the current study could not be diagnosed prospectively in all cases on precontrast T1 and T2 scans images. Postcontrast scans were most efficacious for diagnosis and lesion delineation, with high-dose (0.3 mmol/kg) scans superior to standard dose (0.1 mmol/kg). PMID- 9342118 TI - Computed tomography characterization of renal cell tumors in correlation with histopathology. AB - RATIONALE AND OBJECTIVES: The authors distinguish the histomorphologic subtypes of renal cell tumors (RCTs) by computed tomography (CT). METHODS: In a consensus conference between radiologists, pathologists, and urologists, the CT criteria of the various subtypes of RCTs (clear cell, chromophilic cell, chromophobic cell renal carcinoma and oncocytoma) were established. Computed tomography scans of 65 resected RCTs were reevaluated independently by seven radiologists. Using a numerical scoring system, they first attempted to differentiate clear cell from nonclear cell RCTs. A further attempt then was made to classify each tumor into one of the four categories. RESULTS: The sensitivity for the diagnosis of clear cell RCT was 72.5% (213 of 294 true-positive findings) and 82% (132 of 161 true positive findings) for the nonclear cell group. For tumors more than 3 cm in diameter the sensitivities were 80.25% for the clear cell group and 80.7% for the nonclear cell group. Specific differentiation into the four subtypes was not possible. Oncocytomas were classified correctly in only 6 of 49 observations (12.2%). CONCLUSIONS: Small clear cell tumors often fail to show the CT characteristics that would permit an accurate classification. In tumors measuring 3 cm or more, differentiation between clear cell and nonclear cell types by means of CT criteria is possible. Nevertheless, as RCTs show a great variation in appearance, a differentiation into subtypes of the nonclear cell RCTs cannot be accomplished by CT. Using a uniform examination protocol and spiral scanning technique, the sensitivity of CT in the diagnosis of the subtypes of RCTs may be able to be further increased. Some tumors, especially oncocytomas, undoubtedly will remain diagnostic dilemmas. PMID- 9342119 TI - Magnetic resonance imaging of Achilles tendon in patients with rheumatoid arthritis. AB - RATIONALE AND OBJECTIVES: The authors characterize the appearance of the Achilles tendon in patients with rheumatoid arthritis and differentiate this appearance from degenerative tendinopathy in patients with chronic pain of the heel using magnetic resonance (MR) imaging. METHODS: Thirty patients with rheumatoid arthritis and 28 patients with chronic pain of the heel underwent MR imaging of the ankle and foot. Three radiologists independently assessed the MR images with respect to size, shape, and intratendinal signal characteristics of the Achilles tendon. The Achilles tendon was considered abnormal on MR imaging when intratendinous signal alterations or an anteroposterior measurement greater than 8 mm was seen. Physical examination of the Achilles tendons was accomplished in both groups. Operation confirmed the diagnosis of 13 patients in the second group with chronic pain of the heel. RESULTS: The Achilles tendon of 83% of patients with rheumatoid arthritis demonstrated various intratendinous patterns (longitudinal, reticular, nodular) of intermediate signal intensity on all pulse sequences on MR imaging. Ninety percent of patients with rheumatoid tendinopathy showed no enlargement of the anteroposterior diameter of the Achilles tendon. In addition, all patients with rheumatoid arthritis had findings compatible with an inflammation of the retrocalcaneal bursa on MR imaging, whereas none of the patients with tendinopathy associated with chronic heel pain had retrocalcaneal bursitis. All patients, however, had enlargement of the anteroposterior diameter of the Achilles tendon. Seventy-nine percent showed various intratendinous lesions of intermediate signal intensity on all pulse sequences. Twenty-one percent of patients had an enlargement of the Achilles tendon without intratendinous changes. CONCLUSIONS: Rheumatoid tendinopathy can be distinguished from degenerative tendinopathy in patients with chronic pain of the heel with MR imaging. Inflammation of the retrocalcaneal bursa and the absence of enlargement of the tendon combined with the presence of intratendinous signal alterations are characteristic findings of rheumatoid tendinopathy. PMID- 9342120 TI - Influence of soft tissue (fat and fat-free mass) on ultrasound bone velocity: an in vivo study. AB - RATIONALE AND OBJECTIVES: The authors determine the relative effect of soft tissue compartments, body fat (percent [%Bfat] and weight [Bfat kg]) and fat-free mass (FFM kg), on measurements of ultrasound bone velocity (UBV m/second). METHODS: The authors measured UBV in proximal phalanxes and body fat and fat-free mass by near infrared interactance in 40 healthy premenopausal women (mean age +/ standard deviation 28.2 +/- 3.8 years). RESULTS: Correlation study (Fisher's r to z) showed that UBV correlated negatively with %Bfat (r = -0.61, P < 0.0001), Bfat kg (r = -0.56, P = 0.0001) and marginally with body weight (r = -0.33, P = 0.0403), but did not correlate with FFM kg or H2O L (both r = -0.08, P not significant). When the correlation test was adjusted for weight and age (partial correlation), the negative correlation between UBV and %Bfat persisted (r = 0.54, P < 0.0005; and r = -0.63, P < 0.0001, respectively) and the correlation with FFM kg, adjusted for weight, became positive and significant (r = 0.55, P < 0.0005). CONCLUSIONS: These results, to our knowledge, are the first to be obtained by in vivo evaluation of UBV in relation to body fat and fat-free mass. Body fat, but not fat-free mass, was the main factor affecting UBV. This confirms the deficiency of UBV measurements, considering that obesity is a protective factor for bone mass. PMID- 9342121 TI - Prospective evaluation of bone marrow signal changes on magnetic resonance tomography during high-dose chemotherapy and peripheral blood stem cell transplantation in patients with breast cancer. AB - RATIONALE AND OBJECTIVES: The authors evaluate bone marrow signal changes on magnetic resonance (MR) imaging during high-dose chemotherapy and peripheral blood stem cell transplantation (PBSCT). METHODS: Fourteen patients with breast cancer without bone metastases underwent four sagittal MR imaging studies with T1 weighted, T2-weighted turbo spin-echo and inversion recovery sequences with short inversion time (STIR) of the lumbar spine: (1) during initial staging, (2) prior to high-dose chemotherapy after two cycles of induction chemotherapy, (3) early after PBSCT with a leukocyte exceeding 2000/microL, and (4) 6 to 8 weeks after PBSCT. Signal intensity ratios of averaged lumbar bone marrow to nucleus pulposus were measured and homogeneity was evaluated visually using a semiquantitative score. RESULTS: Compared with the initial finding (1): Signal intensity ratios were altered significantly at (2): T1-weighted: -22% +/- 14, P < 0.001; T2 weighted: -11% +/- 11, P < 0.01; STIR: +33% +/- 31, P < 0.01; Signal intensity ratios were altered significantly at (3): T2-weighted: -23% +/- 12, P < 0.001, STIR: -22% +/- 23, P < 0.01; and Signal intensity ratios were altered significantly at (4): only STIR: -16% +/- 19, P < 0.01. Homogeneity scores decreased at (3) for T1-weighted and STIR sequences (-1.6 +/- 0.5 to -2.0 +/- 0.7, P < 0.01 and -1.0 +/- 0.5 to -1.4 +/- 0.5, P < 0.01, respectively) and at (4) for the latter sequence (-1.0 +/- 0.5 versus -1.4 +/- 0.5, P < 0.01). At (4), T1-weighted images were less homogenous than initially in 3 of 14 (21%) patients. CONCLUSIONS: Magnetic resonance imaging demonstrates significant alterations of bone marrow composition during PBSCT but allows differentiation of benign therapy related changes from those known in metastatic disease after completion of PBSCT. PMID- 9342122 TI - Comparative cytotoxicity of low- and high-osmolar contrast media to human fibroblasts and rat mesangial cells in culture. AB - RATIONALE AND OBJECTIVES: The authors investigate the relative sensitivity of rat mesangial cells to iodinated contrast media (CM) and control solutions versus less differentiated cells (ie, human fibroblasts) and compare the effects of low osmolar ionic (ioxaglate) and nonionic (iopamidol) and high-osmolar ionic (diatrizoate) CM on rat mesangial cells. METHODS: The cytotoxic effects of ioxaglate and control solutions of sodium chloride and mannitol were assessed by neutral red uptake in isolated rat mesangial cells and human fibroblasts. In a second series of studies, the cytotoxic effects of ioxaglate, iopamidol, and diatrizoate (0 to 100 mg I/mL) on rat mesangial cells were compared. RESULTS: Rat mesangial cells were more sensitive to the cytotoxic effects of ioxaglate than the less differentiated human fibroblasts between 70 and 100 mg I/mL. A similar discrepancy was observed in the case of control solutions, sodium chloride, and mannitol. Ioxaglate and iopamidol induced a similar level of cytotoxicity in rat mesangial cells whereas the high-osmolar agent diatrizoate was significantly more cytotoxic. However, the calculated inhibitory concentrations of 50% of all three CM were associated with similar osmolalities, suggesting a major role for this parameter in the case of such media. CONCLUSIONS: Rat mesangial cells are more sensitive to the cytotoxic effects of CM and hyperosmolar solutions than the less differentiated human fibroblasts. High-osmolar CM are more cytotoxic than ionic and nonionic low-osmolar CM to rat mesangial cells. Ionicity seems to play no deleterious role at similar iodine concentrations because ioxaglate and iopamidol had equivalent cytotoxic effects on mesangial cells. PMID- 9342123 TI - Effect of microsphere size distribution on the ultrasonographic contrast efficacy of air-filled albumin microspheres in the left ventricle of dog hearts. AB - RATIONALE AND OBJECTIVES: The in vitro ultrasonographic contrast efficacy of air filled albumin microspheres has been found to depend on the size distribution of microspheres. The objective of the current study was to empirically describe the relationship between the size distribution of injected air-filled albumin microspheres and the in vivo contrast efficacy after lung capillary filtration in a dog model. METHODS: Twenty different air-filled microspheres with large and well-defined differences in size distribution were prepared from nine different batches of Albunex (Molecular Biosystems Inc.) and subsequently characterized by Coulter counting. The in vivo ultrasonographic contrast enhancement of these preparations was investigated with a VingMed CFM750 in closed chest model in six mongrel dogs. The observed contrast efficacy, measured as gray-level enhancement in the left ventricle (LV), was correlated to the microsphere size distribution, using both univariate and multivariate approaches. RESULTS: The results demonstrated a significant contribution to LV contrast efficacy from microspheres larger than approximately 7 microm, and a lack of contribution from microspheres smaller than approximately 7 microm. Linear relationships were found between LV contrast efficacy, and both the number concentration of microspheres between 8 to 12 microm and the total microsphere volume concentration. No significant covariance between in vivo contrast efficacy and the number concentration between 1 to 38 microm or 4 to 10 microm was observed. The multivariate model showed a significant contribution to the in vivo gray-level enhancement from microspheres in the size range 7 to 15 microm, with optimal efficacy per microsphere at approximately 13 microm. CONCLUSIONS: Large microspheres (> 7 microm), which had been expected to be trapped in the lung capillary bed, contribute most of the observed ultrasound contrast in the LV of the heart. PMID- 9342124 TI - Coil on the loop and selectively enhanced stiffness for improved control: feasibility study in the neonatal ovine model of the large patent ductus arteriosus. AB - RATIONALE AND OBJECTIVES: Several devices have been suggested and tested for interventional closure of the persistent ductus arteriosus. Coils were found effective only in small ducts due to their lack of maneuverability and physical limitation of grip forces leading to risk of embolization. The authors evaluated the feasibility to place single coils with selectively enhanced stiffness into high shunting ductus arteriosus, the coils being deployed and controlled through a veno-arterial loop in a bovine model. METHODS: "Double-cone" shaped, stainless steel coils with enhanced stiffness of the outer rings were mounted on either end on a nitinol core wire. A snap-in mechanism attaches the coil to this delivery wire and is freed by a pusher system of coiled steel wire that is advanced over the core wire. Forming a veno-arterial loop across the patent ductus allows for repositioning into the pulmonary artery or aortic catheter. A chronic lamb model of large patent ductus arteriosus (PDAs) (> or = 9 mm) was used in which ductus patency was secured by a protocol of repetitive angioplasties. Different systems (n = 10) were placed having retrieved the previous coil by a snare after definitive release. RESULTS: Placement of coils was possible in all 10 attempts. The coils were pulled back into the catheters between five and eight times before definitive release. CONCLUSIONS: The device allows controlled placement of single coils in our model of large PDAs and may lead the way to overcome previous limitations of coils. Clinical trials are warranted. PMID- 9342125 TI - Dynamic magnetic resonance imaging in the diagnosis of groin hernia. AB - RATIONALE AND OBJECTIVES: The authors determine the feasibility of dynamic magnetic resonance (MR) imaging in the diagnosis of groin hernia. METHODS: Ten volunteers and 10 patients with clinically evident and surgically proven herniations were evaluated using T1-, and T2-weighted sequences and two dynamic sequences. The visibility of anatomic structures that are crucial for the assessment and the differentiation of inguinofemoral herniations was evaluated. RESULTS: The inguinal rings could be identified in all subjects. The inferior epigastric vessels could be identified in 85%. In 10 patients, 11 hernias were found at MR imaging, whereas at surgery and physical examination 13 herniations were diagnosed (84.6%). The two hernias that were missed initially could be identified retrospectively on MR imaging. One volunteer showed a small bilateral inguinal hernia on MR imaging that could be confirmed on physical examination. CONCLUSIONS: The anatomic structures that are crucial for the assessment and the differentiation of inguinofemoral herniations can be identified prospectively with MR imaging. PMID- 9342126 TI - Effects of radiographic contrast media. PMID- 9342127 TI - Cardiovascular risk during ECT: managing the managers. PMID- 9342128 TI - The mortality rate with ECT. AB - ECT is a low-risk procedure, even in the older cardiac patient who is fast becoming the modal candidate for this therapy. To put the mortal risk with ECT in proper perspective, it is only necessary to note that ECT is about 10 times safer than childbirth, that approximately 6 times as many deaths annually in the U.S. are caused by lightning as by ECT, that two complications of psychotropic drug therapy in younger women-fatal myocardial infarction and fatal subarachnoid hemorrhage-virtually never occur with ECT, and that the death rate reported for ECT is an order of magnitude smaller than the spontaneous death rate in the general population. PMID- 9342129 TI - Diagnosis and management of ischemic heart disease in the patient scheduled to undergo electroconvulsive therapy. AB - The cardiovascular risk of electroconvulsive therapy (ECT) is a product of the stress of ECT itself and the severity and stability of coronary artery disease (CAD), as well as other cardiovascular factors. ECT itself represents a relatively low-risk procedure. Patient-specific risk can be defined by a combination of clinical evaluation and noninvasive testing, much of which is aimed at detecting the presence and staging the severity and stability of CAD. Patients at high risk of a cardiac complication include those with severe or unstable symptoms of CAD, and they should undergo extensive cardiac evaluation before ECT Patients at low risk likely need no further evaluation and can undergo ECT. Patients at intermediate risk should have careful clinical evaluation, and most likely noninvasive evaluation, which should include some form of stress testing. Medical therapy should be continued and/or maximized in all patients with CAD. It is expected that with careful screening, patients with established CAD can undergo ECT safely. PMID- 9342130 TI - Electroconvulsive therapy in patients with heart failure or valvular heart disease. AB - As the use of electroconvulsive therapy (ECT) increases, the chance of a practitioner's encountering a patient with significant heart failure, ventricular dysfunction, or valvular heart disease also increases. This article reviews the epidemiology, pathophysiology, and available data on the risk of ECT in these patients. Recommendations are made regarding evaluation and treatment of such patients. Some special situations are identified that may require a modification of routine procedures. Overall, ECT can be performed safely in most patients with underlying cardiac conditions, as long as appropriate precautions are taken to identify these patients ahead of time. PMID- 9342131 TI - Anesthetic considerations of cardiovascular risk during electroconvulsive therapy. AB - This article focuses on anesthetic considerations of cardiovascular risk for electroconvulsive (ECT) therapy. Preoperative evaluation, intraoperative management, and postoperative care are reviewed. Although the anesthetic risk to ECT patients is quite low, elderly patients or those presenting with known cardiovascular disease may be at increased risk and need special intervention or management during ECT. PMID- 9342132 TI - The effect of esmolol on ST-segment depression and arrhythmias after electroconvulsive therapy. AB - Electroconvulsive therapy (ECT) induces sympathetically mediated hemodynamic alterations that can be associated with myocardial ischemia and arrhythmia generation. Esmolol, a short-acting beta-blocker, blunts the hypertension and tachycardia seen with ECT. The purpose of this study is to determine whether esmolol use during ECT reduces the incidence of myocardial ischemia or arrhythmias after ECT. In a randomized, double-blind, placebo-controlled protocol, with each patient acting as his/her own control, the effects of esmolol on the incidence of myocardial ischemia and arrhythmias were studied using two lead Holter monitoring for at least 2 h post-ECT. Nineteen patients underwent 71 ECT treatments (34 placebo, 37 esmolol), recording 746 h of Holter data. The esmolol group had significantly reduced heart rate and mean arterial pressure immediately after ECT. There was no difference in the incidence of ECG defined ischemia post-ECT between groups, with 7 of 19 (36.8%) patients in the esmolol group showing ST-segment depression compared with 5 of 19 (26.3%) in the placebo group. There was no difference between groups in arrhythmia detection. This experiment demonstrates that (a) ECT is associated with a significant incidence of ST-segment depression, (b) esmolol blunts the sympathetic discharge during ECT, and (c) esmolol does not reduce the incidence of post-ECT ischemia or arrhythmia. PMID- 9342133 TI - Effect of esmolol pretreatment on EEG seizure morphology in RUL ECT. AB - Intravenous beta-blockers are an effective means of controlling heart rate and blood pressure during electroconvulsive therapy (ECT), but have been shown to decrease seizure duration. While the importance of seizure duration to the antidepressant response of ECT grows less certain, there is growing evidence that seizure morphology predicts the antidepressant effect of ECT. This study examined the impact of esmolol pretreatment on seizure morphology. Eighteen depressed patients (6 men, 12 women; 69 +/- 12.8 years old) received ECT with and without esmolol pretreatment in a randomized, blinded crossover design. The seizures were blindly rated for duration of motor convulsion, duration of electroencephalogram (EEG) seizure, degree of seizure regularity, and degree of postictal EEG suppression. Esmolol shortened the duration of the motor convulsion and degraded the quality of the ictal regularity. Routine administration of intravenous esmolol before ECT may cause a decrease in ictal regularity. Careful consideration should be given to the potential benefits of esmolol versus the deleterious effect on the electrophysiologic process. Esmolol may still be indicated on a case-by-case basis for extreme tachycardia or hypertension associated with ECT, and presumably poses no problem for the therapeutic effect of ECT if given after the seizure is over. PMID- 9342134 TI - Postictal neurogenic pulmonary edema: experience from an ECT model. AB - Neurogenic pulmonary edema (NPE) is thought to rarely occur after seizures. Paradoxically, NPE is frequently found (>80%) at autopsy in epileptic patients who die unexpectedly. The reason for the discrepancy between the frequency of NPE found at autopsy and that after uncomplicated seizures is unclear. The literature suggests that subclinical NPE occurs rarely after uncomplicated seizures and resolves within a few hours, but is undetected because of infrequent use of routine chest radiographs early after a seizure occurs. This pilot study examined the frequency of subclinical, radiographically confirmed NPE after electroconvulsive therapy (ECT)-induced seizures. If shown to occur, ECT-induced subclinical NPE would provide an easily reproducible model to study NPE after seizures in patients with epilepsy. Given that sudden unexplained death syndrome accounts for approximately 10% of the deaths in patients with epilepsy, an easily reproducible model for NPE would have heuristic value. We examined 12 patients undergoing ECT for depression with chest radiographs before and after ECT. In this group, only 1 of the 12 patients had subclinical NPE in their post-ECT radiograph. We conclude that subclinical NPE does not significantly occur after seizures in patients undergoing ECT and therefore, would not serve as an application for research. PMID- 9342136 TI - Electroconvulsive therapy in patients with aortic stenosis. AB - Aortic stenosis confers an increased risk of complications during procedures with general anesthesia. There are no previously reported cases of electroconvulsive therapy (ECT) in patients with this valvular defect. Two cases are described of patients with moderate to severe aortic stenosis confirmed by echocardiography in whom courses of ECT resulted in clinical improvement without untoward cardiac complications. It is concluded that ECT can be safely given to patients with aortic stenosis in whom left ventricular function is normal. PMID- 9342135 TI - Evaluation of pre-ECS antihypertensive drug administration in the attenuation of ECS-induced retrograde amnesia. AB - Two once-daily electroconvulsive shocks (ECS) produced retrograde amnesia in rats trained on a Hebb-Williams maze; Verapamil (12.5 mg/kg, i.p.) or felodipine (10 mg/kg, p.o.) administered half an hour before each ECS attenuated this ECS induced amnesia. Hence, these drugs may hold promise for the containment of amnesia induced by electroconvulsive therapy (ECT). Speculatively, one or more of several mechanisms may be involved: cerebral vasodilatation, enhancement of cholinergic tone, and inhibition of calcium-mediated impairment of neuronal function. These drugs may also act by attenuating the systolic surge in blood pressure during ECT, thereby decreasing edema due to cerebral hyperperfusion, as well as decreasing the possible transfer of potentially neurotoxic macromolecules through a putative breach in the blood-brain barrier. PMID- 9342137 TI - Safe administration of ECT in a patient with a cardiac aneurysm and multiple cardiac risk factors. AB - A 41-year-old man with a left ventricular aneurysm received electroconvulsive therapy (ECT) for treatment of depression, tolerating the procedure without any cardiovascular complications. This case suggests that ECT may be safely administered to patients with ventricular aneurysm and multiple cardiac risk factors provided that additional precautions are taken. PMID- 9342138 TI - Residual colours: a proposal for aminochromography. PMID- 9342139 TI - Robust multivariate statistics and the prediction of protein secondary structure content. PMID- 9342140 TI - N-terminal truncation mutagenesis of equinatoxin II, a pore-forming protein from the sea anemone Actinia equina. AB - The role of the N-terminal segment 1-33 of equinatoxin II, a 20 kDa pore-forming protein from the sea anemone Actinia equina, was studied by N-truncation mutagenesis. A part of this segment was classified as being amphiphilic and membrane seeking. Wild-type equinatoxin II and its mutants lacking 5, 10 and 33 amino acid residues, respectively, were produced in Escherichia coli using T7 RNA polymerase-based expression vector. Soluble recombinant proteins were isolated from bacterial lysates and assayed for their inhibition by sphingomyelin, binding to red blood cells and hemolytic activity. The N-terminal deletion of 33 amino acids resulted in an insoluble protein, while mutants lacking 5 and 10 residues expressed increased relative avidity for sphingomyelin and red blood cell membranes. Their specific hemolytic activity was decreased, however, with increasing truncation. The results suggest that the N-terminus, which has been found to be conserved in sea anemone pore-forming toxins, contributes to the solubility of the equinatoxin II, but it is not essential for binding to lipid membranes. It is very likely that the N-terminus play a role in the formation of functional pores. PMID- 9342141 TI - Relations of the numbers of protein sequences, families and folds. AB - The relations among the numbers of protein sequences, families and folds have been studied theoretically. It is found that the number of families is related to the natural logarithm of the number of sequences. The logarithmic relation should not be changed regardless of what value of the homology threshold is applied in the protein sequence comparison routines. To study the relation between the numbers of families and folds, the degenerate degree of a fold has been introduced. The degenerate degree of a fold is the number of protein families which adopt the same fold. The distribution of the degenerate degrees of folds has been found to be very likely exponential. Based on the distribution, the average degenerate degree d is calculated. The number of folds is simply equal to that of families divided by the average degenerate degree of folds. It is shown that d is an increasing function of time. The current value of d is about 2. It will continue to increase and reach the value of at least 3.3 in some years. By using the above result, the numbers of protein folds for four species have been estimated. In particular, the number of folds for human proteins is estimated to be < or =5200. PMID- 9342142 TI - Prediction of protein supersecondary structures based on the artificial neural network method. AB - The sequence patterns of 11 types of frequently occurring connecting peptides, which lead to a classification of supersecondary motifs, were studied. A database of protein supersecondary motifs was set up. An artificial neural network method, i.e. the back propagation neural network, was applied to the predictions of the supersecondary motifs from protein sequences. The prediction correctness ratios are higher than 70%, and many of them vary from 75 to 82%. These results are useful for the further study of the relationship between the structure and function of proteins. It may also provide some important information about protein design and the prediction of protein tertiary structure. PMID- 9342143 TI - Exploring the limits of nearest neighbour secondary structure prediction. AB - This paper presents a simple and robust secondary structure prediction scheme (SIMPA96) based on an updated version of the nearest neighbour method. Using a larger database of known structures, the Blosum 62 substitution matrix and a regularization algorithm, the three state prediction accuracy is increased by 4.7 percentage points to 67.7% for a single sequence and up to 72.8% when using multiple alignments. The increase in prediction accuracy with respect to the previous version can be almost entirely ascribed to the sevenfold increase in the size of the database. A more detailed analysis of the results shows that badly predicted regions of a protein sequence are randomly distributed throughout the database and that the goal of perfect secondary structure predictions by methods which use only local sequence information is illusory. PMID- 9342144 TI - Correlation between side chain mobility and conformation in protein structures. AB - Thermal factors of protein atoms as determined by X-ray crystallographic techniques show a tendency to be larger in side chains with unfavourable local conformations rather than in those displaying conformational energy minima. It follows that side chain atoms are more mobile if they are in a non-rotameric configuration and that the stereochemistry of protein structures cannot be fully assessed or simulated without consideration of thermal factors that monitor flexibility in various regions of the protein. The observations should also prove useful in protein folding and design. PMID- 9342145 TI - Modeling protein stability: a theoretical analysis of the stability of T4 lysozyme mutants. AB - Free energy calculations were conducted to determine the relative stability of the unnatural amino acid mutants of T4 lysozyme norvaline (Nvl) and O-methyl serine (Mse) and of alanine at residue 133, which is leucine in the native sequence. These calculations were performed both to assess the validity of the methodology and to gain a better understanding of the forces which contribute to protein stability. Peptides of different length were used to model the denatured state. Restraints were employed to force sampling of the side chain chi1 dihedral of the perturbed side chain, and the effect of protein repacking in response to mutation was studied through the use of different constraint sets. In addition, the convergence behavior and hysteresis of the simulations in the folded and unfolded states were determined. The calculated results agree well with experiment, + 1.84 versus + 1.56 kcal/mol for Mse-->Nvl and -3.48 versus -2.2 to 3.6 kcal/mol for Nvl-->Ala. We find that free energy calculations can provide useful insights to protein stability when conducted carefully on a well chosen system. Our results suggest that loss of packing interactions in the native state is a major source of destabilization for mutants which decrease the amount of buried nonpolar surface area and that subtle responses of the backbone affect the magnitude of the loss of stability. We show that the conformational freedom of the chi1 dihedral has a noticeable effect on protein stability and that the solvation of amino acid side chains is strongly influenced by interactions with the peptide backbone. PMID- 9342146 TI - Improving pKa calculations with consideration of hydration entropy. AB - Continuum dielectric modelling of electrostatics interactions in macromolecules provides a valuable tool in the study of structure-function relationships, but falls short of providing consistently accurate calculated pKas. It is suggested that the model can be significantly improved with the inclusion of a term that estimates the entropy associated with first hydration shell solvent ordering, with reference to computed results for cysteines in DsbA and thioredoxin, and aspartic and glutamic acids in a number of proteins. The modification is based on the geometry of charge burial and an hydration number, which is adjustable (by fit to experiment), and is uniform within each class of ionizable group studied. The potential for further development is clear within this framework, since experiment and simulation can furnish non-adjustable, ionizable group-specific, hydration numbers. PMID- 9342147 TI - Effects of single-residue substitutions on negative cooperativity in ligand binding to dihydrofolate reductase. AB - The effects of six amino acid substitutions in Lactobacillus casei dihydrofolate reductase, predominantly in the coenzyme binding site, on catalysis and on the negative cooperativity between NADPH and tetrahydrofolate binding have been determined. Replacement of Leu62, His64 or Leu54 by alanine has no effect on kcat, and produces only modest changes in negative cooperativity. Alanine substitution of His77, which interacts indirectly with the coenzyme adenine ring, leads to a doubling of the negative cooperativity and a consequent doubling of kcat. Replacement of Arg43, which interacts with the coenzyme 2'-phosphate, by alanine, or of Trp21, which interacts with the coenzyme nicotinamide ring, by histidine leads to a 20-100-fold decrease in negative cooperativity. In both mutants there is a decrease in kcat; isotope effects show that product release is largely rate-limiting in R43A, whereas in W21H hydride ion transfer is rate limiting. 1H NMR has been used to obtain information on the extent of the structural changes produced by the substitutions. This varies from very local effects in H64A to very widespread effects in W21H. These changes are used as the basis for discussion of the mechanisms of the functional effects of the various substitutions. It is suggested that residues in helix C, beta-strand 3 and the beta3-beta4 loop may be involved in the transmission of effects between the coenzyme and substrate binding sites. PMID- 9342148 TI - Identification of amino acid residues of abrin-a A chain is essential for catalysis and reassociation with abrin-a B chain by site-directed mutagenesis. AB - Abrin is a toxic protein consisting of two subunits, an enzymatic A chain (ABRaA) and a lectin-active B chain (ABRaB), linked by a disulfide bond. Site-directed mutagenesis was performed using PCR to study how the conserved amino acid residues, Tyr74, Tyr113, Glu164 and Trp198, around the active site of ABRaA are involved in enzyme catalysis, enzyme-substrate recognition and reassociation of ABRaA with ABRaB. The protein biosynthesis inhibitory activities of Y74F, Y113F and W198F were decreased moderately to that of wild type reABRaA, while that of E164Q decreased dramatically. Kinetic analysis showed that the kat of Y74F, Y113F and W198F resembled that of wild type, while the Km increased significantly. W198F did not reassociate with ABRaB to form heterodimers, while Y74F, Y113F and E164Q did. SDS-PAGE analysis of ABRaA treated with trypsin showed that reABRaA, Y74F, Y113F and E164Q survived digestion, whereas W198F was not protected from digestion. CD spectra revealed that W198F showed significant conformational changes. These observations suggest that E164 is directly involved in catalysis, and Tyr74, Tyr113 and Trp198 in substrate binding, while Trp198 also plays an important role in maintaining the conformation of ABRaA required for its reassociation with ABRaB. PMID- 9342149 TI - Heavy chain CDR3 optimization of a germline encoded recombinant antibody fragment predisposed to bind the U1A protein. AB - Previously, we described a DP-65 encoded heavy chain variable (VH) gene restriction in anti-U1A antibodies. The U1A protein (a component of the U1 ribonucleoprotein particle) is an important autoantigenic target in certain systemic lupus erythematosus (SLE) patients. Here we examined the effect of randomizing amino acids in the heavy chain complementarity determining region 3 (CDR3) of this germline encoded recombinant antibody fragment on binding to the U1A protein. A phage display library was constructed using the DP-65 VH domain with four randomized CDR3 residues and our results showed that a high frequency (10%) of the randomized mutants in the unselected library were able to bind the U1A protein. This corroborates our previous finding that this VH domain provides an appropriate structure for U1A binding, although the nature of the CDR3 residues appears crucial in determining whether or not this VH domain binds U1A. After two rounds of selection U1A binders show a consensus sequence in their randomized CDR3 residues i.e. S(K,R,S)XG, in which X is an uncharged residue. This consensus is partially present in an antibody which was derived from an SLE patient indicating that this consensus, to some extent, is also followed in vivo. Clones which match the consensus sequence obtained up to 25-fold higher affinities compared with the original clones, illustrating the importance of the VH CDR3 residues in determining the affinity of these antibodies. PMID- 9342150 TI - Postpolio muscular dysfunction: relationships between muscle energy metabolism, subjective symptoms, magnetic resonance imaging, electromyography, and muscle strength. AB - Eleven patients with previous polio were studied. The concentration of energy related metabolites and energy charge was measured from the vastus lateralis muscle, as was isometric muscle strength of knee extension. Cross-sectional area of the quadriceps femoris muscle was calculated from magnetic resonance imaging. Reinnervation was studied using macroelectromyography. Muscle weakness, pain, and newly acquired muscle weakness in the legs was estimated by the patients. The findings in the legs in which the patients experienced new loss of muscle function were compared with the stable legs. There were no significant differences between these groups in any of the objectively measured variables. Only hip pain correlated with new loss of muscle function. Creatine phosphate was decreased in 5 patients. The symptoms and subjective muscle strength did not correlate with any of the objective measurements. There were no significant relationships between energy-related metabolites and postpolio symptoms. PMID- 9342151 TI - A 31P-magnetic resonance spectroscopy and biochemical study of the mo(vbr) mouse: potential model for the mitochondrial encephalomyopathies. AB - 31P-magnetic resonance spectroscopy (31P-MRS) provides new biochemical information on mitochondrial disorders affecting brain and muscle. To elucidate the mechanisms of mitochondrial abnormalities, however, animal models are needed. We assessed the mo(vbr) (mottled viable brindled) mouse for its value in studying (1) energetics of a mitochondrial disorder and (2) 31P-MRS changes associated with mitochondrial abnormalities in vivo. The maximal activity of succinate cytochrome c reductase was significantly reduced in mo(vbr) muscle compared to controls, whereas cytochrome oxidase activity was only reduced in mo(vbr) brain. 31P-MRS of mo(vbr) brain showed an increased pH, but no changes in any metabolite ratios. The phosphocreatine (PCr) recovery rate after exercise was reduced in muscles from mo(vbr) mice, indicating impairment of oxidative metabolism. We conclude that mo(vbr) brain and muscle tissue have biochemical abnormalities consistent with mitochondrial impairment. The PCr recovery rate, measured by 31P MRS, was sensitive to the muscle abnormality. This strain is best described as having chronic mitochondrial dysfunction. PMID- 9342152 TI - Diagnostic yield of noninvasive high spatial resolution electromyography in neuromuscular diseases. AB - High Spatial Resolution electromyography (HSR-EMG), a new kind of noninvasive surface EMG based on a spatial filtering technique, was evaluated with respect to the diagnosis of neuromuscular diseases. HSR-EMG measurements were recorded from 61 healthy subjects and 72 patients with different neuromuscular diseases and analyzed quantitatively. The results indicate that a few parameters such as muscular conduction velocity, dwell time over root mean square, autocorrelation function, and chi-value are sufficient to recognize and classify specific signal alterations due to neuromuscular disorders. A diagnostic evaluation procedure calculating automatically the most probable diagnosis from the parameter results could assign the correct diagnosis to about 81% of the investigated patients and healthy subjects. Myopathic disorders were recognized with a sensitivity of 85% (specificity: 97%), neuropathic disorders with a sensitivity of 68% (specificity: 98%). We conclude that HSR-EMG shows a diagnostic validity similar to that described in literature for needle EMG. Moreover, the noninvasive technique provides the advantage of a simple and painless application. PMID- 9342153 TI - Acute corticosteroid myopathy in intensive care patients. AB - Several recent studies have attributed the occurrence of acute myopathy in intensive care unit patients to the combination of corticosteroids and neuromuscular junction blocking agents (NMBAs) used for mechanical ventilation. We present 4 patients who developed acute myopathy after administration of high doses of glucocorticoids during sedation with propofol without any NMBAs. All patients had elevated creatine kinase levels. Electrophysiological studies indicated normal motor and sensory nerve conduction velocities but reduced motor nerve response amplitudes. Needle electromyography identified abnormal spontaneous activity; motor unit potentials were polyphasic of low amplitude and short duration, characteristic of a myopathic process. Muscle biopsy demonstrated a prominent acute necrotizing myopathy in all 4 patients with a loss of thick filaments. Our observations support glucocorticoids rather than NMBAs as the main offending drug in acute corticosteroid myopathy. The predisposing factor should be the hypersensitivity of paralyzed muscles to corticosteroids regardless of the drug inducing paralysis: NMBAs or propofol. PMID- 9342154 TI - Comparison of single-fiber and macro electrode recordings: relationship to motor unit action potential duration. AB - The factors contributing to the duration of a motor unit action potential (MUAP) are believed to be well known, with both manual measurements and computer simulations agreeing with respect to MUAP durations approaching 10 ms. In this investigation, it is clearly demonstrated that use of a wide-open amplifier bandpass combined with signal-to-noise ratio enhancement results in MUAP durations approaching 30 ms recorded with either a macro or single-fiber electrode. Why the clinically recorded MUAP duration differs significantly from these physiologic durations is discussed. A hypothesis is presented whereby the major contributing factor toward MUAP duration is the total time of action potential transmembrane current flow along the muscle fiber from end-plate zone to musculotendinous junction. PMID- 9342155 TI - Effects of cutaneous histamine application in patients with sympathetic reflex dystrophy. AB - Thirty-six patients suffering from acute reflex sympathetic dystrophy (RSD) were examined in order to evaluate nociceptive C-fibers. Axon reflex vasodilatation was induced by iontophoresis of histamine and recorded (laser Doppler flux). The strength of concomitant sensation was rated on a visual analogue scale, and the quality was characterized as itching or burning pain. Skin temperature was recorded by infrared thermography. The results were compared with investigations of unaffected limbs of patients and volunteers. The histamine-induced sensation on the symptomatic side was more often burning pain than itching (P < 0.001), and skin temperature was increased on the affected limb (P < 0.001). Axon reflex vasodilatation and the strength of sensations were unaltered. In conclusion, this study rules out a significant deterioration of afferent C-fibers in RSD, but gives evidence of sensitization of nociceptive function. This nociceptive sensitization has to be taken into consideration for effective treatment of RSD. PMID- 9342156 TI - Value of sphincter electromyography in the diagnosis of multiple system atrophy. AB - It is clinically important, to distinguish between idiopathic Parkinson's disease (IPD) and multiple system atrophy (MSA) not only because of the implications for prognosis but also because urinary incontinence is often an early troublesome feature of MSA and by making the correct neurological diagnosis inappropriate urological surgery may be avoided. Onuf's nucleus in the sacral cord is the location of the anterior horn cells innervating the sphincters, and it is among central nervous system sites affected by neuronal cell loss in MSA but not in IPD. A systematic analysis of motor units recorded from the sphincter looking for changes of chronic reinnervation has therefore been used to distinguish between these conditions. Sphincter electromyography (EMG) was carried out in 126 patients with suspected MSA with review of their case notes up to 2 years later. Of those in whom a diagnosis of MSA was made, 82% had had an abnormal sphincter EMG. PMID- 9342157 TI - Partial transformation from fast to slow muscle fibers induced by deafferentation of capsaicin-sensitive muscle afferents. AB - Mechanical and histochemical characteristics of the lateral gastrocnemius (LG) muscle of the rat were examined 21 days after capsaicin injection into the LG muscle. The capsaicin caused a decrease in generation rate of twitch and tetanic tension and an increase in fatigue resistance of LG muscle. The histochemical muscle fiber profile evaluated by myosin adenosine triphosphatase and reduced nicotinamide adenine dinucleotide tetrazolium reductase methods showed an increase of type I and IIC fibers and a decrease of the type IIB in whole muscle, and a decrease of the IIA, IIX fibers in the red part accompanied by their increase in the white part. Therefore the capsaicin treatment, which selectively eliminated fibers belonging to the III and IV groups of muscle afferents, induced muscle fiber transformation from fast contracting fatiguing fibers to slowly contracting nonfatiguing ones. PMID- 9342158 TI - Directional sensibility for quantification of tactile dysfunction. AB - Examination of tactile directional sensibility, i.e., the ability to tell the direction of an object's motion across the skin, has been recommended by several authors for examination of patients with somatosensory disorders. Recent findings about the physiological mechanisms underlying directional sensibility suggested possibilities to further improve the test. In the present investigation a test was constructed that allowed a semiquantification of the directional sensibility of six body areas within 20 min. Normal values were obtained by testing healthy subjects (n = 40), and the normal values were compared to those obtained in a group of patients with tactile symptoms (n = 20). Ten of the patients had abnormal sensory conduction in one or several nerves, and they also had abnormal directional sensibility. Hence, examination of directional sensibility, according to the present protocol, provides a semiquantitative test that appears to be as sensitive as electrophysiological measurement of conduction in detecting dysfunction in tactile nerves. PMID- 9342159 TI - Skeletal muscle fiber degeneration in mdx mice induced by electrical stimulation. AB - We present an in vitro model in which mouse skeletal muscle fibers undergo degeneration by increasing the current strength of tetanic stimulation. To understand the mechanisms of muscle fiber necrosis in Duchenne muscular dystrophy patients, the process of fiber degeneration was compared between mdx and control mice. The process consisted of four steps, beginning with muscle fiber contraction and extending to onset of myofibril disruption. The four processes were not observed in fibers in Krebs-HEPES (-Ca2+) buffer, nor in the presence of L-type Ca2+ channel blockers. These results suggest that this degenerative phenomenon is regulated by intracellular Ca2+, which moved into fibers mainly through voltage-dependent L-type Ca2+ channels. With the exception of myofibril disruption, mdx mice also exhibited the three other steps, but at a significantly lower current strength than in the fibers in the control mice. We postulate that excess Ca2+ flux occurs in fibers, mainly through abnormal L-type Ca2+ channels, and that the excessively accumulated calcium results in premature degeneration of the fibers by tetanic contraction. This study would provide a clue to investigate and prevent the degeneration processes in Duchenne muscular dystrophy. PMID- 9342160 TI - Motor involvement in acute herpes zoster. AB - Motor involvement in acute herpes zoster is considered rare, but its incidence is unknown. In a sample of 40 patients with acute herpes zoster of varying severity, an abnormal electromyogram (EMG) (fibrillation, positive waves, high-frequency discharges) was found in 21 (53%), suggesting extension of inflammation to the anterior horn and/or anterior motor roots. In the majority of patients these changes were not confined to the segment invaded by the rash but were widespread, extending several segments cranially and caudally, and both ipsi- and contralaterally. In 5 (13%) patients these changes became more extensive on repeat EMG over a period of months. There was no association between severity of rash, pain, postherpetic neuralgia, and EMG changes. We conclude that widespread subclinical motor involvement is relatively common in herpes zoster, may last for months, and is easily detectable by EMG. PMID- 9342161 TI - The effect of pharmacologic erection on the dorsal nerve of the penis. AB - The dorsal nerve of the penis (DNP) is the primary source of afferent somatic input from the penis and is critical in the male sexual functions of erection and ejaculation. Using genitourinary electrodiagnostic techniques, this study was conducted to investigate the effect of pharmacologic erection on the DNP. Three tests were administered, and the changes in the DNP between flaccid, stretched, and erect states were examined. Calculated nerve conduction velocity (cNCV) measurements of the DNP increased with pharmacologic erection because mechanical straightening of the nerve allowed for a more precise measurement of nerve length. The latencies of the cortical evoked response and the bulbocavernosus reflex were not significantly changed with stretch or pharmacologic erection. In the evaluation of impotence, cNCV DNP measurements should be performed on the erect penis. PMID- 9342162 TI - n-Hexane neuropathy due to rubber cement sniffing. AB - n-Hexane neuropathy has been described after glue sniffing and industrial exposure. Onset may be subacute and reminiscent of Guillain-Barre syndrome. Although the primary pathology is axonal, electrophysiologic evaluation is frequently most remarkable for conduction slowing. We describe a patient with a severe subacute neuropathy following n-hexane exposure via glue sniffing. Although symptoms worsened after termination of exposure ("coasting"), strength gradually improved to near normal. Sources of toxic exposure should be sought in all patients with subacute demyelinating neuropathies. PMID- 9342163 TI - Four new polymorphisms in the human dystrophin gene from an Argentinian population. AB - Duchenne muscular dystrophy and its allelic disorder Becker muscular dystrophy are among the most common hereditary human pathologies (1:3500). Two thirds of the genomic alterations responsible for these diseases involve gross gene rearrangements such as deletions, and less frequently duplications. The remaining one third includes point mutations such as deletions, insertions, and substitutions. This study describes four nonpreviously reported polymorphisms in the dystrophin gene by using the polymerase chain reaction/single-strand conformation polymorphism technique and subsequent nonisotopic silver staining. PMID- 9342164 TI - POEMS syndrome in a 24-year-old man associated with vitamin B12 deficiency and a solitary lytic bone lesion. AB - We report one of the youngest cases of POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes), presenting in a 24-year-old man. A solitary lytic bone lesion and vitamin B12 deficiency were also found. We suggest that this syndrome be considered in cases of subacute polyneuropathy developing in young adults. PMID- 9342165 TI - Myasthenia gravis and Charcot-Marie-Tooth disease type 1A: an unusual combination of diseases. AB - Concurrence of myasthenia gravis (MG) and Charcot-Marie-Tooth type 1 (CMT1A) neuropathy is rare. We describe a 60-year-old woman with MG and genetically proved CMT1A. The fluctuating ocular symptoms and proximal limb weakness were markedly improved by pyridostigmine treatment. Recognition of the possible association of MG and CMT1A in the same patient is important because the therapeutic result is rewarding. PMID- 9342166 TI - Median-ulnar anastomosis to thenar, hypothenar, and first dorsal interosseous muscles: collision technique confirmation. AB - Median-ulnar anastomosis (Martin-Gruber anastomosis; MGA) is traditionally diagnosed based upon changes in compound muscle action potential (CMAP) amplitude following proximal stimulation. We describe a rare patient with a MGA innervating thenar, hypothenar, and first dorsal interosseous muscles. Proximal stimulation, however, evoked CMAPs with striking changes in morphology and area but only minimal amplitude changes, due to concomitant diagnoses of carpal tunnel syndrome and polyneuropathy. Collision studies were therefore required for diagnostic confirmation of the MGA. PMID- 9342167 TI - Magnetic resonance imaging of skeletal muscles in the polymyositis. AB - Magnetic resonance imaging of skeletal muscles was performed in 11 patients with polymyositis. Two types of muscle lesions were revealed. The first, inflammation, showed increased signal intensity on T2-weighted images and iso-intensity on T1 weighted images. The second, fatty replacement, showed increased signal intensity on both images. The coronal sections could elucidate the extension of the lesion in each affected muscle. Inflammation was relatively diffuse, while homogeneous fatty replacement tended to begin at the lower myotendinous junctions. PMID- 9342168 TI - Azathioprine hypersensitivity reactions: caution upon rechallenge. PMID- 9342169 TI - Neuroselective current perception threshold quantitative sensory test. PMID- 9342170 TI - Overview: pharmacokinetic drug-drug interactions. PMID- 9342171 TI - Role of cytochrome P450 enzymes in drug-drug interactions. AB - Many adverse drug-drug interactions are attributable to pharmacokinetic problems and can be understood in terms of alterations of P450-catalyzed reactions. Much is now known about the human P450 enzymes and what they do, and it has been possible to apply this information to issues related to practical problems. A relatively small subset of the total number of human P450s appears to be responsible for a large fraction of the oxidation of drugs. The three major reasons for drug-drug interactions involving the P450s are induction, inhibition, and possibly stimulation, with inhibition appearing to be the most important in terms of known clinical problems. With the available knowledge of human P450s and reagents, it is possible to do in vitro experiments with drugs and make useful predictions. The results can be tested in vivo, again using assays based on our knowledge of human P450s. This approach has the capability of not only improving predictions about which drugs might show serious interaction problems, but also decreasing the number of in vivo interaction studies that must be performed. These approaches should improve with further refinement and technical advances. PMID- 9342172 TI - The liver as a target for chemical-chemical interactions. PMID- 9342173 TI - Application of human liver microsomes in metabolism-based drug-drug interactions: in vitro-in vivo correlations and the Abbott Laboratories experience. PMID- 9342174 TI - Primary hepatocyte cultures as an in vitro experimental model for the evaluation of pharmacokinetic drug-drug interactions. PMID- 9342175 TI - Liver slices as a model in drug metabolism. PMID- 9342176 TI - Use of cDNA-expressed human cytochrome P450 enzymes to study potential drug-drug interactions. AB - Complementary DNA (cDNA)-expressed human cytochrome P450 enzymes provide a reproducible, consistent source of single enzymes for many types of studies. The use of single enzymes systems, relative to multienzyme systems, has distinct advantages and disadvantages depending on the specific application. cDNA expressed materials have advantages in the analysis of cytochrome P450 form selective metabolism of a drug or drug candidate. This analysis can be accomplished by direct incubation of the drug with microsomes prepared from cells expressing a single cytochrome P450 form coupled with analysis of either metabolite formation or loss of parent compound. This approach allows the unambiguous assignment of specific biotransformations to specific enzymes. However, extending these data to the balance of enzymes present in human liver microsomes can be problematic. New approaches for relating rates of metabolism for cDNA-expressed enzymes to human liver microsomes metabolism are being developed (Crespi, 1995). In addition, cDNA-expressed enzymes can be used to study the cytochrome P450 form-selective inhibition by drugs or drug candidates. This analysis is accomplished through the study of the inhibition of the metabolism of a model substrate by the drug or drug candidate. Through these analyses, apparent Ki values can be obtained and compared to Ki values for known, clinically significant inhibitors of the same enzyme. For this application, cDNA expressed, single enzyme systems have distinct advantages because of greater flexibility in the choice of model substrates and the lack of competing pathways of metabolism. Specific data for the use of cDNA-expressed CYP2C9, CYP2D6, and CYP3A4 are presented. PMID- 9342177 TI - Pharmacokinetics of drug interactions. PMID- 9342178 TI - Experimental models for evaluating enzyme induction potential of new drug candidates in animals and humans and a strategy for their use. AB - Experimental models that have application for evaluating enzyme induction potential have been described in order of increasing complexity. The main focus was on models that have had wide application thus far. However, many new models are currently being developed that may have future applications in evaluating enzyme induction potential. A strategy to evaluate the enzyme induction potential of drug candidates was outlined. This scheme uses a combination of new and established techniques to evaluate data in a stepwise manner that is appropriate to the drug's current stage of development. PMID- 9342179 TI - Metabolic drug-drug interactions: perspective from FDA medical and clinical pharmacology reviewers. PMID- 9342180 TI - Drug interactions: perspectives of the Canadian Drugs Directorate. PMID- 9342181 TI - Overview of experimental approaches for study of drug metabolism and drug-drug interactions. PMID- 9342182 TI - Phenotypes of c-Myc-deficient rat fibroblasts isolated by targeted homologous recombination. AB - Rat fibroblast cell lines with targeted disruptions of both c-myc gene copies were constructed. Although c-myc null cells are viable, their growth is significantly impaired. The absence of detectable N-myc or L-myc expression indicates that Myc function is not absolutely essential for cell viability. The c myc null phenotype is stable and can be reverted by introduction of a c-myc transgene. Exponentially growing c-myc null cells have the same cell size, rRNA, and total protein content as their c-myc +/+ parents, but the rates of RNA and protein accumulation as well as protein degradation are reduced. Both the G1 and G2 phases of the cell cycle are significantly lengthened, whereas the duration of S phase is unaffected. This is the first direct demonstration of a requirement for c-myc in G2. The G0-->S transition is synchronous, but S-phase entry is significantly delayed. The c-myc null cell lines reported here are a new experimental system in which to investigate the importance of putative c-Myc target genes and to identify novel downstream genes involved in cell cycle progression and apoptosis. PMID- 9342183 TI - Nuclear factor-kappaB/Rel blocks transforming growth factor beta1-induced apoptosis of murine hepatocyte cell lines. AB - Treatment of hepatocytes with transforming growth factor beta1 (TGF-beta1) induces growth arrest, which is followed by extensive cell death by apoptosis. Previously, we found that TGF-beta1 down-modulates nuclear factor (NF)-kappaB/Rel activity in murine B cell lymphomas, inducing apoptosis. Furthermore, p65 (RelA) deficient mice died during gestation due to apoptosis of liver cells. Here we have explored the effects of TGF-beta1 on hepatocytes, using two untransformed murine hepatocyte cell lines, AML-12 and NMH, which constitutively express classical NF-kappaB. TGF-beta1 treatment caused increased NF-kappaB binding that was followed by a dramatic decrease in NF-kappaB levels that preceded apoptosis. Ectopic c-Rel expression ablated apoptosis induced by TGF-beta1. The down regulation in NF-kappaB activity correlated with elevated IkappaB-alpha expression due to hypophosphorylation and increased IkappaB-alpha protein stability. Thus, NF-kappaB factor expression acts directly to promote liver cell survival. Furthermore, these findings characterize a novel signaling pathway for TGF-beta1 in epithelial cells involving down-regulation of NF-kappaB/Rel factors activity through posttranslational modification of IkappaB-alpha protein. PMID- 9342184 TI - Osteopontin expression in mammary gland development and tumorigenesis. AB - Osteopontin (OPN) is a secreted phosphoprotein that binds to cells via an Arg-Gly Asp sequence and to mineralized surfaces. The protein can mediate cell adhesion and is strongly implicated in transformation and tumorigenesis. We have examined the expression pattern of OPN in mouse mammary glands at different stages of postnatal development. Whereas OPN is expressed at low-to-moderate levels in mammary glands from virgin and pregnant mice, the levels of OPN mRNA are extremely high in the lactating gland, consistent with the presence of the protein in milk. Expression is highest at 2 days of lactation and declines thereafter, but it remains high through involution. OPN expression is restricted to small nests or groups of cells at 9 days of involution. These results suggest that OPN may play a specific role in the process of involution that may be distinct from its role during lactation. In mammary tumors arising spontaneously in transgenic mice expressing the oncogenes c-myc and/or v-Ha-ras under the control of the mouse mammary tumor virus promoter, the level of OPN expression is increased dramatically over that in the normal gland in these same animals. Numerous cells expressing OPN mRNA are widespread throughout the tumors. OPN protein is detectable by Western blotting in extracts from the mammary gland at 2 days of lactation and from the tumors, but not in mammary glands at other stages of development. We hypothesize that OPN is exported from most tissues and that the protein is only detectable in tissues elaborating fluids, such as the lactating mammary gland, or in pathological situations when expression of OPN is abnormally high, such as in tumors. PMID- 9342185 TI - Retinoid-mediated inhibition of cell growth with stimulation of apoptosis in aggressive B-cell lymphomas. AB - Retinoids have been shown to modulate cell growth and differentiation in a variety of human tumor cell types, but their effects on B-cell non-Hodgkin's lymphomas (NHL-B) have not been explored. In this study, all-trans retinoic acid (ATRA) in the free form and liposome-encapsulated form (L-ATRA) were used to determine effects on fresh NHL-B patient cells as well as cell lines recently established from both HIV-negative and -positive NHL-B patient biopsies. Both ATRA and L-ATRA were found to inhibit cell proliferation in NHL-B cells. However, L-ATRA was found to be superior to free ATRA in inhibiting cell proliferation of NHL-B cells and resulted in greater than 90% cell growth inhibition in a dose dependent manner. In addition, L-ATRA also induced high levels of apoptosis in NHL-B cells in vitro. To delineate the apoptotic pathways involved, the expression of the apoptosis suppressor oncogene bcl-2 was evaluated in different NHL-B cells with and without the t(14;18) chromosomal translocation. After L-ATRA exposure, more than a 50% reduction in the expression of bcl-2 protein was observed. bcl-2 message levels were also down-regulated in the L-ATRA-sensitive NHL-B cells. Bax protein levels were analyzed and found to be up-regulated in L ATRA-sensitive NHL-B cells. Similar results were observed in sensitive AIDS/lymphoma cell lines. Experiments using an RAR-alpha antagonist (RO 41-5253) showed that both the proliferation inhibition and apoptosis induced by L-ATRA could be blocked in NHL-B cells. The findings of the present study indicate that L-ATRA may possess therapeutic potential in blocking cell proliferation, inducing apoptosis, and PMID- 9342186 TI - Monocytic differentiation of HL-60 promyelocytic leukemia cells correlates with the induction of Bcl-xL. AB - Treatment of human promyelocytic leukemia HL-60 cells with phorbol esters ultimately induces the differentiation of these cells along the monocyte/macrophage lineage, whereas treatment with retinoic acid or DMSO induces granulocytic/neutrophillic differentiation. In this study, we demonstrate the disparate fates of HL-60 cells treated with the phorbol ester 12,13-phorbol dibutyric acid (PDBu) or DMSO. After DMSO treatment, HL-60 cells eventually died via apoptosis, whereas the viability of PDBu-treated cells was not affected during the same interval. The levels of the apoptosis effector proteins Bak and Bad were enhanced, whereas there was a slight down-regulation of the apoptosis suppressor protein Bcl-2 after treatment of the cells with PDBu and DMSO. Treatment with DMSO resulted in the elevation of the apoptosis effector Bax, whereas treatment with PDBu did not significantly alter the levels of this protein. However, treatment of HL-60 cells with PDBu induced the rapid expression of the apoptosis suppressor protein Bcl-xL, whereas the expression of this protein remained unaltered in DMSO-treated cells. The generality of this finding was confirmed by the induction of Bcl-xL in human myeloid U-937 cells, human peripheral blood monocytes exposed to phorbol ester, and mouse thioglycollate activated and resident peritoneal macrophages. PDBu-treated HL-60 cells remained viable for 7 days and thereafter began to die via apoptosis, with a concomitant down-regulation of Bcl-xL. In conclusion, we propose that Bcl-xL expression is associated with differentiation and survival of hematopoietic cells along the monocyte/macrophage lineage. PMID- 9342187 TI - Dexamethasone signaling is required to establish the postmitotic state of adipocyte development. AB - The clonal expansion phase of 3T3-L1 adipose conversion is a distinct mitotic period during which the initiation of differentiation occurs concomitant with a discrete set of mitotic divisions. During clonal expansion, a cocktail of adipogenic hormones, including the glucocorticoid dexamethasone, induced 3T3-L1 cells to progress from postconfluent adipoblasts to postmitotic adipocytes. It is reported here that expression of the growth arrest-associated gene 2 (gas2) discriminated reversible, postconfluent growth arrest from irreversible, postmitotic growth arrest. In the absence of dexamethasone, 3T3-L1 cells underwent mitoses but failed to establish postmitotic growth arrest, as evidenced by the persistence of elevated GAS2 mRNA. Moreover, the dexamethasone-deprived 3T3-L1 cells appeared to revert to postconfluent growth arrest, as judged by (a) their ability to reenter logarithmic growth under permissive conditions, and (b) their ability to undergo adipose conversion when subsequently challenged with a complete cocktail of adipogenic hormones. The growth potentiating factor-encoding gene, gas6, was shown to be an immediate-early target of dexamethasone. These findings reveal the requirement for dexamethasone in establishing the postmitotic state that accompanies differentiation. In addition, the results suggest that dexamethasone signaling influences the mitotic divisions of clonal expansion by determining the nature of the growth arrest state assumed upon exit from the cell cycle. PMID- 9342188 TI - Differential regulation of transcription of p21 and cyclin D1 conferred by TAF(II)250. AB - The TATA-binding protein-associated factor TAF(II)250, the largest subunit of TFIID, was first identified as the cell cycle regulatory protein CCG1. The ts13 Syrian hamster ovary fibroblast cell line, which contains a temperature-sensitive point mutation in TAF(II)250/CCG1, is arrested in G1 following a shift to the nonpermissive temperature. Here we demonstrate that the level of the D-type cyclins, in particular D1, was reduced, whereas the level of the cyclin-dependent kinase inhibitor p21 was stimulated in ts13 cells at the nonpermissive temperature. The levels of expression of cyclins A and E were not affected by temperature shift. We further show that at least part of the regulation of D1 and p21 levels in ts13 cells is mediated at the level of transcription initiation. These results suggest that the effect of the temperature-sensitive mutation TAF(II)250 on cell growth can be mediated through the differential regulation of transcription of specific cell growth regulatory genes, such as cyclin D1 and p21. PMID- 9342189 TI - Inactivation of G2 checkpoint function and chromosomal destabilization are linked in human fibroblasts expressing human papillomavirus type 16 E6. AB - Chromosomal stability was linked to G2 checkpoint function in human fibroblasts expressing the human papillomavirus type 16 E6 oncoprotein. Soon after expression of E6, cells displayed an undamaged, diploid karyotype and normal mitotic delay after gamma-irradiation. As the E6-expressing cells aged through their in vitro life span, G2 checkpoint function diminished progressively. After 30-70 population doublings, 60-86% of the E6 cells displayed defective G2 checkpoint response. This attenuation of G2 checkpoint function was also associated with radiation-resistant cyclin B1/CDK1 protein kinase activity. Numerical and structural abnormalities of chromosomes developed in unirradiated E6 cells with kinetics that mirrored the loss of G2 checkpoint function. A significant correlation between inactivation of the G2 checkpoint and acquisition of chromosomal abnormalities was found, suggesting that the G2 checkpoint represents a barrier to genetic instability in cells lacking G1 checkpoint function. PMID- 9342191 TI - The tumor suppressor gene p53 can mediate transforming growth [corrected] factor beta1-induced differentiation of leukemic cells independently of activation of the retinoblastoma protein. AB - Although the involvement of the tumor suppressor gene p53 in normal hematopoiesis is uncertain, it can give rise to differentiation signals in leukemic cells. It is not clear, however, whether differentiation merely is a consequence of the ability of p53 to arrest cell proliferation or whether hitherto unknown molecular mechanisms are responsible for the p53-mediated differentiation. To further explore the role of p53 in leukemic cell differentiation, we investigated whether transforming growth factor beta1 (TGF-beta1), a cytokine involved in cell cycle control at several levels, can cooperate with wild-type p53 to induce differentiation of monoblastic U-937 and erythroleukemic K562 cells. Indeed, wild type p53-expressing cells were found to be more sensitive to TGF-beta1-induced differentiation than control cells, lending support to the idea that p53 is of importance for differentiation induction of leukemic cells. In addition, it is shown that TGF-beta1 can suppress p53-mediated cell death, thus reinforcing the differentiation response. The cyclin-dependent kinase inhibitor p21 and the retinoblastoma protein (pRb) are downstream effectors of p53-mediated growth arrest. Therefore, the roles for these molecules in p53-mediated differentiation were examined. The p53-dependent signals of differentiation were associated with induction of p21 in both cell lines investigated. However, activation of pRb by induced hypophosphorylation and concomitant decreased growth rate on p53-mediated differentiation was observed only in U-937 cells expressing an inducible, temperature-sensitive form of p53 but not in K562 cells constitutively expressing p53. Thus, our data suggest a role for p53 in the regulation of differentiation in leukemic cells that can be independent of its ability to activate pRb and arrest cell proliferation. PMID- 9342190 TI - The homeobox-containing Engrailed (En-1) product down-regulates the expression of Pax-6 through a DNA binding-independent mechanism. AB - By in situ hybridization of quail neuroretinas, we observed that Engrailed (En-1) is expressed both in the ganglionic and the amacrine cell layers, similarly to Pax-6. Because we observed a decrease of Pax-6 expression in the neuroretina of hatched animals, we studied the effect of the chicken En-1 and En-2 proteins on Pax-6 expression. En-1 and to some extent En-2 were able to repress the basal and the p46Pax-6-activated transcription from the two Pax-6 promoters. Infection of retinal pigmented epithelium by a virus encoding the En-1 protein repressed the endogenous Pax-6, and a similar effect was observed with a homeodomain-deleted En 1. In vitro interaction indicates that En proteins are able to interact with the p46Pax-6 through the paired domain. This interaction negatively regulates the DNA binding properties of the p46Pax-6. These results suggest an interplay between En 1 and Pax-6 during the central nervous system development and indicate that En-1 may be a negative regulator of Pax-6. PMID- 9342192 TI - Novel cystatin B mutation and diagnostic PCR assay in an Unverricht-Lundborg progressive myoclonus epilepsy patient. AB - Two mutations in the cystatin B gene, a 3' splice mutation and a stop codon mutation, were previously found in patients with progressive myoclonus epilepsy of Unverricht-Lundborg type [Pennacchio et al. (1996): Science 271:1731-1734]. We present here a new mutation 2404deltaTC: a 2-bp deletion within the third exon of the cystatin B gene in an Unverricht-Lundborg patient. This mutation results in a frameshift and consequently premature termination of protein synthesis. Complete sequencing of the coding region and splice junctions of the cystatin B gene showed that neither of the two previously known mutations was present in this patient. The level of cystatin B mRNA in an immortalized cell line was found to be decreased, as had been reported for other Unverricht-Lundborg patients. The new mutation further supports the argument that defects in the cystatin B gene cause the Unverricht-Lundborg form of progressive myoclonus epilepsy. We describe a simple PCR method which can detect the 2404deltaTC deletion. This assay, together with previously described PCR assays for the other two known mutations, should prove useful in confirming clinically difficult diagnoses of Unverricht Lundborg disease. PMID- 9342194 TI - Genetic association between monoamine oxidase and manic-depressive illness: comparison of relative risk and haplotype relative risk data. AB - There have been several conflicting reports of association of monoamine oxidase (MAO) A gene polymorphisms and bipolar affective disorder. In order to determine the possible role of the MAO region in susceptibility to affective disorders in an independent sample, we have genotyped 83 probands of bipolar affective disorder families, 56 sets of parents of bipolar probands, and 84 normal controls for intronic simple sequence repeat polymorphisms of the MAO-A and MAO-B genes. For MAO-A there were no significant differences in allele frequencies between bipolar and normal control groups for both genders. However, for MAO-B there were significant differences between groups for both genders. In contrast, allele-wise haplotype relative risk analysis for the 56 bipolar proband-parent trios found no significant differences between transmitted and non-transmitted allele frequencies for MAO-A or B. These data do not support the association of MAO-A or B with bipolar affective disorder but do demonstrate that undetected population stratification can be an important source of bias in case-control studies. PMID- 9342193 TI - Absence of linkage for schizophrenia on the short arm of chromosome 5 in multiplex Canadian families. AB - A VNTR for the human dopamine transporter gene (DAT-1) has been localized to chromosome 5p15.3. Silverman et al. [1996] found evidence for genetic linkage of the D5S111 locus, located just centromeric to DAT-1, to schizophrenia and related disorders in a large Hispanic family. We evaluated five markers on 5p, including D5S111 and the DAT-1 VNTR, in five multiplex schizophrenic families, assuming autosomal dominant transmission (subjects assessed n = 122, DNAs available n = 96, individuals with schizophrenia and schizoaffective disorder n = 36, broader spectrum disorders n = 14). LOD scores were negative across all families for all markers tested, and overall LOD scores were strongly negative (<-2.0, theta = 0) across all five families for each of the markers typed. Thus, there is no evidence to support the linkage of markers in this region of chromosome 5 to schizophrenia in this sample of families. PMID- 9342195 TI - Human dopamine transporter gene polymorphism (VNTR) and alcoholism. AB - The dopamine transporter (DAT1) is responsible for taking released dopamine back up into presynaptic terminals and terminating dopaminergic activity. It has been shown that cocaine binds to the dopamine transporter and blocks dopamine reuptake in a fashion that correlates with cocaine reward and reinforcement. To determine the role of this gene in the development of alcoholism, we have used two approaches, relative risk and haplotype relative risk. The relative risk approach involved 162 alcoholic probands who were categorized into type I and type II, and 89 unrelated normal controls. In the haplotype relative risk approach, 29 trios (father, mother, and proband) were genotyped with dopamine transporter gene polymorphism. Comparison of allele frequencies between total alcoholics, subtypes of alcoholics, and normal controls were negative. The results of haplotype relative risk, differences between alleles transmitted and nontransmitted, were also negative. However, both approaches produced similar results. Therefore, we concluded that the VNTR polymorphism in DAT1 gene is not associated with alcoholism susceptibility genes in our samples. PMID- 9342196 TI - Dopamine D4 receptor exon III alleles and variation of novelty seeking in alcoholics. AB - A dysfunction of dopaminergic neurotansmission has been implicated in alcohol seeking behavior. Recently, a significant association between the seven-repeat allele (DRD4*7R) of a 16 amino acid motif in the third exon of the dopamine D4 receptor gene (DRD4) and the personality trait of novelty seeking has been reported. Our population-based association study tested the hypothesis that the DRD4*7R variant predisposes to high levels of novelty seeking, which may underlie alcohol-seeking behavior. The genotypes of the expressed DRD4 exon III polymorphism were determined in 197 German controls and 252 German alcohol dependent males, of whom 92 alcoholics completed the tridimensional personality questionnaire. We found no significant differences in the DRD4*7R frequencies between controls and alcoholics, including two subgroups (56 alcoholics with dissocial personality disorder according to ICD-10 and 89 alcoholics with severe withdrawal symptoms) with a high level of novelty seeking. The novelty-seeking scores did not differ significantly between alcoholics (including both subgroups) carrying long alleles with six or more repeats compared with those lacking long alleles. The present results do not provide evidence that the DRD4*7R allele contributes a common and relevant effect to alcohol-seeking behavior in our sample of alcoholics. PMID- 9342197 TI - Comparative studies of the CAG repeats in the spinocerebellar ataxia type 1 (SCA1) gene. AB - The CAG repeat tract at the autosomal dominant spinocerebellar ataxia type 1 (SCA1) locus was analyzed in SCA1 families and French-Acadian, African-American, Caucasian, Greenland Inuit, and Thai populations. The normal alleles had 9-37 repeats, whereas disease alleles contained 44-64 repeats. The CAG repeat tract contained one or two CAT interruptions in 44 of 47 normal human chromosomes and in all five chimpanzees examined. In contrast, no CAT interruptions were found in Old World monkeys or expanded human alleles. The number and positions of CAT interruptions may be important in stabilizing CAG repeat tracts in normal chromosomes. At least five codons occupy the region corresponding to the polyglutamine tract at the SCA1 locus in mice, rats, and other rodents. They comprise three or four CCN (coding for proline) in addition to one or two CAG repeats. PMID- 9342198 TI - Monoamine oxidase genes polymorphisms and mood disorder. AB - We investigated a genetic association between mood disorders (bipolar and unipolar) and the alleles of monoamine oxidase (MAO) A and B (MAOA and MAOB). One hundred and twelve unrelated Japanese patients (60 bipolar, 52 unipolar) and 100 controls were genotyped for three markers of MAOA and for one marker of MAOB. No statistically significant difference in the distribution of the alleles existed between cases and controls. Therefore, our results did not support the involvement of the alleles at MAOA and MAOB in the etiology of mood disorder. PMID- 9342199 TI - Ciliary neurotrophic factor null allele frequencies in schizophrenia, affective disorders, and Alzheimer's disease. AB - Ciliary neurotrophic factor (CNTF) is a cytokine that has been reported to affect cellular differentiation. Mouse CNTF knockouts have progressive motor neuron atrophy, but this protein has uncertain physiological function in humans. A naturally occurring CNTF variant in man, observed in many populations, abolishes function of the protein product, providing the opportunity to study loss of CNTF function in humans. It has been reported previously that this variant does not predispose to several neurological and neuropsychiatric disorders, including Alzheimer's disease, but findings have been more ambiguous with respect to other conditions, such as schizophrenia. We report here allele frequencies for this null mutation in populations diagnosed with mood disorders (unipolar depression, single episode or recurrent; N = 59), schizophrenia (N = 66), or Alzheimer's disease (N = 93). We found no association of the CNTF null with any of these phenotypes. There is presently no known phenotype consistently associated with either heterozygosity or homozygosity for the CNTF null allele, suggesting either that this protein does not serve a necessary function in humans or is redundant with some other protein or that any human phenotype associated with absence of CNTF is considerably more subtle than that seen in mouse. PMID- 9342200 TI - Association between dopamine D4 receptor (D4DR) exon III polymorphism and novelty seeking in Japanese subjects. AB - This study was designed to assess the association between novelty seeking and D4DR gene polymorphism in the Japanese population. The 48 bp repeat polymorphism in the third exon of the dopamine D4 receptor gene of 153 normal female students was correlated with personality feature results from the Japanese version of Cloninger's Temperament and Character Inventory. The Novelty Seeking subscale of Exploratory Excitability had a significant association with long alleles of the polymorphic exon III repeat sequence of D4DR. Our results suggest that there is an association between long alleles of the polymorphic exon III repeat sequence of D4DR and the personality traits of the Novelty Seeking subscale of Exploratory Excitability, regardless of racial differences in the frequencies of D4DR exon III repeat polymorphism. PMID- 9342201 TI - Association analysis of the 5-HT2C receptor and 5-HT transporter genes in bipolar disorder. AB - We selected 42 patients with bipolar disorder type I (BPI) and 40 healthy controls for genetic analysis of DNA polymorphisms in the serotonin receptor 2c (5-HTR2c) and serotonin transporter (5-HTT) genes. No significant associations were found in the total patient sample. However, when the individuals were divided according to gender, trends for association with both polymorphisms (P = 0.051 for 5-HTR2c and P = 0.049 for 5-HTT) in female patients were observed. These results suggest that variations in these genes may be responsible for a minor increase in susceptibility for bipolar disorder in women. PMID- 9342202 TI - Reduced subcortical brain volumes in nonpsychotic siblings of schizophrenic patients: a pilot magnetic resonance imaging study. AB - Substantial evidence suggests that nonpsychotic relatives of schizophrenia patients manifest subtle abnormalities in communication, eye movements, event related potentials, and neuropsychological processes of attention, reasoning, and memory. We sought to determine whether adult relatives without psychosis or schizophrenia spectrum diagnoses might also have structural brain abnormalities, particularly in subcortical regions found to be impaired in patients with schizophrenia itself. Subjects were six sisters of schizophrenic patients and eleven normal female controls. Sixty contiguous 3 mm coronal, T1-weighted 3D magnetic resonance images (MRI) of the entire brain were acquired on a 1.5 Tesla magnet. Cortical and subcortical gray and white matter was segmented using a semiautomated intensity contour mapping algorithm. Volumes were adjusted for total brain volumes. Adjusted gray matter subcortical volumes were significantly smaller in relatives than in controls in total hippocampus, right amygdala, right putamen, left thalamus, and brainstem. Relatives had significantly enlarged left and total inferior lateral ventricles. These results, though preliminary, suggest that some never-psychotic relatives of schizophrenic patients have abnormal brain structure. If replicated in a larger sample including both sexes, these results would suggest that the genetic liability to schizophrenia is also expressed as structural brain abnormalities. PMID- 9342203 TI - A recognisable behavioural phenotype associated with terminal deletions of the short arm of chromosome 8. AB - We report the clinical findings in 5 patients with a terminal deletion of the short arm of chromosome 8. Mild developmental delay was constantly present, in association with microcephaly in 4 of 5 patients. Facial anomalies were mild or absent. A congenital heart defect was present in 3 patients: an atrioventricular septal defect (AVSD) in 2 and an atrial septal defect type II (ASDII) with pulmonary stenosis in one. A highly similar pattern of behavioural difficulties was present in the 3 older children (8-11 years), with outbursts of aggressiveness and destructive behaviour. Follow-up in one patient showed that at the age of 16 years, these behavioural problems had largely disappeared. This observation suggests that in addition to mental retardation, microcephaly, congenital heart defect (typically AVSD), a terminal deletion of chromosome 8p may be associated with a characteristic behavioural phenotype during childhood. PMID- 9342204 TI - Cognitive, adaptive, and behavioral characteristics of Williams syndrome. AB - Williams syndrome is a genetic disorder linked to cognitive and behavioral patterns of varying consistency; this study was conducted to clarify further the strengths and weaknesses of children with Williams syndrome. Fifteen subjects with the characteristic features of Williams syndrome were evaluated using the Stanford-Binet Intelligence Scale for Children, Fourth Edition; the Vineland Adaptive Behavior Scales, Interview Edition; and the Child Behavior Checklist. Cognitive skills ranged from the Moderate Range of Mental Retardation to the Low Average range, with relative strengths in nonverbal and quantitative reasoning. Adaptive skills were delayed, with strengths in communication and socialization. Behaviorally, clinically significant levels of attention problems, borderline significant levels of social and thought problems, and significantly low levels of social contacts and structured activities were found. In contrast to the findings of many other studies of Williams syndrome, language skills and short term memory skills were weak. Children with Williams syndrome may present a more evenly developed intellectual profile, with verbal and nonverbal skills being commensurate. In conclusion, a variety of cognitive, adaptive, and behavioral patterns have been shown to be possible in Williams syndrome; therefore, a single predictable cognitive or behavioral phenotype cannot be assumed. PMID- 9342205 TI - Methylenetetrahydrofolate reductase variant and schizophrenia/depression. AB - Patients with methylenetetrahydrofolate reductase (MTHFR) deficiency often show psychiatric manifestations. Since a common variant of the MTHFR gene, T677(Ala), responsible for the thermolabile MTHFR with less than 50% specific MTHFR activity, has been reported, we examined whether the T677 allele is associated with psychiatric disorders in an unrelated Japanese population consisting of 297 schizophrenics, 32 patients with major depression, 40 patients with bipolar disorder, and 419 controls. The genotype homozygous for the T677 allele was significantly frequently observed in schizophrenics with an odds ratio of 1.9 (P = 0.0006), and in patients with major depression with an odds ratio of 2.8 (P = 0.005). Our data suggest associations of the MTHFR gene variant with schizophrenia and depression in the Japanese. PMID- 9342206 TI - Evidence for DYT7 being a common cause of cervical dystonia (torticollis) in Central Europe. AB - Adult-onset focal idiopathic torsion dystonias (AFITD), such as torticollis, have a prevalence similar to that of multiple sclerosis and usually seem sporadic. Only recently has one large AFITD pedigree "K" with autosomal dominant inheritance and reduced penetrance from Northwest Germany provided the opportunity to identify a gene locus on chromosome 18p. We have now tested the relevance of this DYT7 gene locus in a collective of 18 nuclear AFITD families from Central Europe who were genotyped with chromosome 18p microsatellites. In three families, the affected relatives did not share a chromosome 18p haplotype, suggesting locus heterogeneity in AFITD. In the remaining 15 families, significant allelic association was observed for marker D18S1098. This result suggests that DYT7 is a common cause for AFITD at least in Central Europe, that many patients are descended from a common ancestor, and that the DYT7 gene is mapped in a 4.4 centimorgan subregion of chromosome 18p. PMID- 9342207 TI - Thoracic tumors in children with neurofibromatosis-1. AB - Thoracic tumors have been infrequently reported as a complication of neurofibromatosis-1 (NF1). To determine the prevalence and clinical features of thoracic tumors seen in children with NF1, we reviewed medical records and imaging studies for a group of 260 pediatric patients with NF1 followed in a multidisciplinary NF Center. Extrapleural thoracic tumors were seen in nine patients with NF1, corresponding to a prevalence of 3.5% in this hospital-based series of patients. Pathological studies of the tumors demonstrated plexiform neurofibroma in four cases and neurofibrosarcoma in one case. The remaining four cases were suspected to be plexiform neurofibroma based on clinical features but have not been confirmed histologically. Three patients presented with symptoms of chest pain, syncope, or wheezing; six patients were asymptomatic at the time of diagnosis of the tumors. Physical findings frequently found in patients with thoracic tumors were scoliosis (especially focal scoliosis) and visible plexiform neurofibromas of the neck. We conclude that NF1 patients presenting with any of these signs and symptoms should be screened for thoracic tumors with chest X-ray and magnetic resonance imaging as needed. It is unknown whether screening asymptomatic NF1 patients with chest X-rays on a regular basis will result in an improved outcome. PMID- 9342208 TI - Enlarged Sylvian fissures in infants with interstitial deletion of chromosome 22q11. AB - Two infants with chromosome 22q11 deletion syndrome were noted to have symmetrically enlarged Sylvian fissures on cranial MRI. We compared the size of the Sylvian fissures in neuroimaging studies from 17 other subjects with del 22q11 to age-matched disease controls. The mean anterior interopercular distance was used as an index of Sylvian fissure enlargement. Symmetric enlargement of the Sylvian fissures was present in 10 of 17 subjects with del 22q11. The age incidence pattern, as well as follow-up scans in 2 patients, suggests delayed growth of the opercular region in these patients. Subjects with del 22q11 consistently had disproportionate enlargement of the left Sylvian fissure compared to the right. This observation suggests that a gene (or genes) in the deleted region affects the development of the left and right perisylvian cortex in different ways. Abnormal development of the operculum may explain some of the neurodevelopmental features that are common among individuals with 22q11 deletion syndrome. PMID- 9342209 TI - Serotonin transporter gene regulatory region polymorphism and anxiety-related traits in the Japanese. PMID- 9342210 TI - Detection of a large CTG/CAG trinucleotide repeat expansion in a Danish schizophrenia kindred. PMID- 9342211 TI - Bipolar affective disorder, chromosome 16p13.3, and recessive disease genes. PMID- 9342212 TI - The structure, regulation and function of the Janus kinases (JAKs) and the signal transducers and activators of transcription (STATs). AB - Since the discovery of their physiological roles in cytokine signalling, the Janus kinases (JAKs) and the signal transducers and activators of transcription (STATs) have attracted considerable attention, to the point that the concept of a intracellular signalling pathway, named JAK/STAT, has emerged. As originally defined, this pathway involves ligand-dependent activation of a particular class of receptor-associated tyrosine kinases, the JAK proteins, which phosphorylate themselves and receptor components, creating recruitment sites for STAT transcription factors. The STATs are phosphorylated, they dissociate from the receptor x JAK complex and translocate to the nucleus where they participate in transcriptional gene activation. Although this pathway was found initially to be activated by interferons, it is now known that a large number of cytokines, growth factors and hormonal factors activate JAK and/or STAT proteins. Recent findings have suggested that the interdependence of JAKs and STATs might not be absolute as originally thought. PMID- 9342213 TI - Characterisation of the promoter which regulates expression of a phosphoglucomutase-related protein, a component of the dystrophin/utrophin cytoskeleton predominantly expressed in smooth muscle. AB - We have recently characterised a 60-kDa muscle-specific phosphoglucomutase related protein (PGM-RP) which is expressed predominantly in adult visceral and vascular smooth muscle. Here we show that the adult vascular smooth muscle cell line PAC1, which retains the capacity to synthesise metavinculin (a marker of the contractile phenotype) also expressed PGM-RP. However, an embryonic smooth muscle cell line A10, which lacks metavinculin, expressed low levels of PGM-RP. Levels of PGM-RP increased in quiescent PAC1 and A10 cells, and were elevated in response to angiotensin II. PGM-RP is therefore a good marker of the contractile/differentiated smooth muscle phenotype. We have sequenced 1.8 kb of the human PGM-RP promoter and shown that it lacks a conventional TATA box. There are multiple transcription start sites, the most predominant of which are inside an initiator sequence (Inr), which is close to two CT boxes and a GATA element. A minimal promoter-CAT construct (p57-CAT) containing the Inr, a CT box and GATA element directed high-level chloramphenicol acetyltransferase (CAT) expression in the differentiated smooth muscle cell line PAC1, and low-level expression in the embryonic smooth muscle cell line A10. This fits well with the pattern of expression of the endogenous gene. A construct (p146-CAT) containing all of the mRNA initiation sites directed a reduced level of CAT expression, and constructs containing 1.8 kb and 3.3 kb upstream of the major transcription start site displayed even lower activity. Sequence comparisons suggest that the PGM-RP promoter evolved from the main phosphoglucomutase promoter which is active in wide range of cell types. The PGM-RP promoter may have acquired negative regulatory elements as expression of the gene became muscle-specific. PMID- 9342214 TI - Organ-specific activity of the 5' regulatory region of the glutamine synthetase gene in developing mice. AB - Glutamine synthetase (GS) converts ammonia and glutamate into glutamine. We assessed the activity of the 5' regulatory region of the GS gene in developing transgenic mice carrying the chloramphenicol acetyltransferase (CAT) gene under the control of 3150 bp of the upstream sequence of the rat GS gene to obtain insight into the spatiotemporal regulation of its pattern of expression. To determine the organ-specific activity of the 5' regulatory region CAT and GS mRNA expression were compared by ribonuclease-protection and semi-quantitative in situ hybridization analyses. Three patterns were observed: the 5' region is active and involved in the regulation of GS expression throughout development (pericentral hepatocytes, intestines and epididymis); the 5' region shows no activity at any of the ages investigated (periportal hepatocytes and white adipose tissue); and the activity of the 5' region becomes repressed during development (stomach, muscle, brown adipose tissue, kidney, lung and testis). In the second group, an additional element must be responsible for the activation of GS expression. The last group included organs in which the 5' regulatory region is active, but not in the cells that express GS. In these organs, the activity of the 5' regulatory region must be repressed by other regulatory regions of the GS gene that are missing from the transgenic construct. These findings indicate that in addition to the 5' regulatory region, at least two unidentified elements are involved in the spatiotemporal pattern of expression of GS. PMID- 9342215 TI - Characterization of a phosphoprotein whose mRNA is regulated by the mitogenic pathways in dog thyroid cells. AB - We have isolated cDNA clones encoding the dog and human forms of a novel protein whose function is still unknown. Sequence analysis indicates that dog clone c5fw protein contains 343 amino acid residues. several potential phosphorylation sites. and two of the 12 conserved subdomains (VIII and IX) that fold into a common catalytic core structure of the large family of protein kinases. Human clone c5fw shares 95% amino acid identity with its dog counterpart. We have also isolated another human-related clone c5fw sharing 70% amino acid identity with the dog sequence. We transiently expressed c-myc epitope-tagged clone c5fw protein in COS-7 cells and infected thyrocytes in primary culture with a recombinant adenovirus containing clone c5fw cDNA (adenovirus c5fw). In both experiments, a 46-kDa protein was detected and subsequently more extensively characterized. By two-dimensional gel electrophoresis and V8 protease digestion, we showed that this overexpressed protein is phosphorylated on different sites. Moreover, cells stimulated with thyrotropin or epidermal growth factor, thyrotropin and fetal calf serum increased the level of clone c5fw protein produced after infection by adenovirus containing clone c5fw. The disappearance of this 46-kDa protein after 1 h of puromycin treatment indicates that it is a labile protein. Immunofluorescence and subcellular fractionation analysis have revealed that c-myc-tagged clone c5fw was insoluble and localized mainly in the cytoplasm, in the form of granules. PMID- 9342216 TI - Repair pattern in the beta-globin gene cluster of human fibroblasts after ultraviolet irradiation. AB - We have developed a novel technique to determine repair of structurally different DNA lesions. It was used to address the question of whether DNA repair in the absence of transcription occurs in a uniformly random manner or with preferences for certain regions. Human fibroblasts were exposed to ultraviolet light (3-10 J/m2) and treated with 7.5 mM hydroxyurea to inhibit replicative DNA synthesis. During the first hours after irradiation cells were treated with 5 bromodeoxyuridine to label the regions undergoing repair, with the presumption that the regions that have been more efficiently repaired would incorporate more of the nucleoside. A 155-kb DNA sequence containing the entire human beta-globin domain was reconstructed using sequences deposited in the EMBL gene bank. Twelve uniformly long single-copy RNA probes spanning the beta-globin cluster were synthesised in vitro and immobilized on microtiter plates. They were hybridized with DNA from the irradiated cells. The amount of 5-bromodeoxyuridine, incorporated as a result of repair in the DNA fractions hybridized to the different RNA probes, was determined immunochemically using antibody to this nucleoside. By this technique we registered increased repair efficiency in the zone of the permanent scaffold attachment region at the 5'-end of the beta-globin domain during the first hours after ultraviolet irradiation. This result was confirmed and by the more conventional T4 endonuclease V technique detecting the removal of cyclobutane pyrimidine dimers. PMID- 9342217 TI - Characterisation of the rat tissue-type plasminogen activator gene promoter -- identification of a TAAT-containing promoter element. AB - Tissue-type plasminogen activator (tPA) activates plasminogen to the active protease plasmin and is implicated in many biological processes that require extracellular proteolysis. In rat ovarian cells, gonadotropins induce the tPA gene by a cAMP-dependent pathway and this induction correlates with the time of follicular rupture. We have previously identified several promoter elements within the first 621 bp of the rat tPA promoter that are important for constitutive and cAMP-induced expression of the gene, including a cAMP responsive element (CRE), a nuclear factor 1 (NF1) element, a SP1-binding site and a G+C rich box. In this report we have extended our study by analysing promoter constructs, ranging in size from 7.7 kb to 135 bp fused to the luciferase reporter gene. Transient transfection analysis of rat granulosa cells and human 293 cells, reveal that the proximal 268 bp of the promoter is enough to confer high basal and cAMP-induced expression of the gene. At position -162 to -172, between the previously identified CRE and NF1 sites, a novel TAAT-containing promoter element was identified. Mutational inactivation of the TAAT motif indicates that this element is important for both constitutive and cAMP-induced expression of the gene, and for the binding of a presumably novel nuclear factor that we have termed tPA promoter factor-1 (tPF-1). PMID- 9342218 TI - Pathway of detergent-mediated and peptide ligand-mediated refolding of heterodimeric class II major histocompatibility complex (MHC) molecules. AB - We investigated the mechanism of refolding and reassembly of recombinant alpha and beta chains of the class II major histocompatibility molecules (MHC-II) HLA DRB5*0101. Both chains were expressed in the cytosol of Escherichia coli, purified in urea and SDS, and reassembled to functional heterodimers by replacement of SDS by mild detergents, incubation in a redox-shuffling buffer and finally by oxidation and removal of detergent. Refolding was mediated by mild detergents and by peptide ligands. Early stages of structure formation were characterized by circular dichroism, fluorescence, and time-resolved fluorescence anisotropy decay (FAD) spectroscopies. We found that formation of secondary structure was detectable after replacement of SDS by mild detergents. At that stage the alpha and beta chains were still monomeric, the buffer was strongly reducing, and the folding intermediates did not yet interact with peptide ligands. Formation of folding intermediates capable of interacting with peptide ligands was detected after adjusting the redox potential with oxidized glutathione and incubation in mild detergents. We conclude that at that stage a tertiary structure close to the native structure is formed at least locally. The nature and concentration of detergent critically determined the refolding efficiency. We compared detergents with different carbohydrate headgroups, and with aliphatic chains ranging from C6 to C14 in length. For each of the detergents we observed a narrow concentration range for mediating refolding. Surprisingly, detergents with long aliphatic chains had to be used at higher concentrations than short-chain detergents, indicating that increasing the solubility of folding intermediates is not the only function of detergents during a refolding reaction. We discuss structure formation and interactions of detergents with stable folding intermediates. Understanding such interactions will help to develop rational strategies for refolding hydrophobic or oligomeric proteins. PMID- 9342219 TI - Production and structure characterisation of recombinant chromogranin A N terminal fragments (vasostatins) -- evidence of dimer-monomer equilibria. AB - Vasostatins (VS) are vasoinhibitory peptides derived from the N-terminal domain of chromogranin A, a secretory protein present in the electron-dense granules of many neuroendocrine cells. In this work we describe a method for the production in Escherichia coli of large amounts of recombinant vasostatins, corresponding to chromogranin A residues 1-78 (VS-1), and 1-115 (VS-2), and the use of these materials for structure characterisation. The masses of both products were close to the expected values, by SDS/PAGE and mass spectrometry analysis. However, their hydrodynamic behaviours in size-exclusion chromatography corresponded to that of proteins with a larger size. SDS/PAGE analysis of VS-1 and VS-2 after cross-linking with disuccinimidyl suberate indicated that both polypeptides form dimers. VS-2 was almost entirely dimeric at > 4 microM, but rapidly converted to monomer after dilution to 70 nM. The rapid dimer-monomer transition of VS-2 after dilution could be part of a mechanism for regulating its activity and localising its action. Immunological studies of VS-1 have shown that residues 37-70 constitute a highly antigenic region characterised by an abundance of linear epitopes efficiently mimicked by synthetic peptides. The recombinant products and the immunological reagents developed in this work could be valuable tools for further investigating the structure and the function of chromogranin A and its fragments. PMID- 9342220 TI - Functional expression of a tobacco gene related to the serine hydrolase family -- esterase activity towards short-chain dinitrophenyl acylesters. AB - We have recently reported the isolation of a tobacco gene, hsr 203J, whose transcripts accumulate during the hypersensitive reaction, a plant response associated with resistance to pathogens. We present and discuss here some structural and biochemical properties of the gene product. Nucleotide sequence analysis has shown that the hsr 203J gene contains an open reading frame coding for a polypeptide of 335 amino acids. The predicted amino acid sequence contains the GXSXG motif characteristic of serine hydrolases, and displays limited but significant similarity to lipases and esterases of prokaryotic origin. The hsr 203J gene was expressed in Escherichia coli, and the recombinant protein, purified to near homogeneity, was able to degrade p-nitrophenylbutyrate, a general substrate for carboxylesterases. The enzyme was unable to hydrolyze lipids, and was active on short-chain acyl esters only. The hydrolytic activity was abolished by diisopropyl fluorophosphate and a derivative of isocoumarin, as expected for a member of the serine hydrolase family. Sequence similarities between the tobacco esterase and expressed sequence tags in databases suggest the existence of members of this enzyme family in various plant species. PMID- 9342221 TI - 26-hydroxylation of ecdysteroids is catalyzed by a typical cytochrome P-450 dependent oxidase and related to ecdysteroid resistance in an insect cell line. AB - The epithelial cell line from the dipteran Chironomus tentans responds to the insect steroid hormone 20-hydroxyecdysone and the non-steroidal analogue tebufenozide by undergoing a morphogenetic and biochemical differentiation program. Long-term culture in the presence of 20-hydroxyecdysone has resulted in the selection of subclones that are resistant to the steroid but respond normally to the non-steroidal analogue. In the present study, several subclones that were resistant to the steroid hormone have been compared with steroid-sensitive subclones with respect to their capability to metabolize 20-hydroxyecdysone. Homogenates of both types of cells, when incubated with 3H-labelled steroid in the presence of NADPH, producecd 20,26-dihydroxyecdysone, which was further metabolized to two compounds, which behaved less polar than 20-hydroxyecdysone on reverse-phase HPLC. Ecdysone, a less-active hormone precursor, provided 26 hydroxyecdysone as the only product. The metabolites were identified by mass spectrometry coupled to HPLC, chromatography with authentic samples, and formation of acetonides. The structure of 20,26-dihydroxyecydsone was confirmed by 1H-NMR. The enzyme responsible for the synthesis of 20,26-dihydroxyecdysone in the Chironomus cell preparations has been characterized as a typical cytochrome P 450-dependent monooxygenase. It was a strictly microsomal enzyme, sensitive to inhibition by carbon monoxide and imidazole/triazole-based fungicides, and required NADPH for maximal activity. NADH could partly replace NADPH. The Michaelis constant (Km) for 20-hydroxyecdysone was 0.96 microM, and the maximal enzyme velocity (Vmax) was 50 pmol substrate metabolized x mg protein(-1) x min( 1). 26-Hydroxylation of 20-hydroxyecdysone was inhibited by ecdysone, an alternative substrate, and by inokosterone, a product analogue, to 50% at 1.4 microM and 0.73 microM, respectively. When various subclones were compared with respect to their in vitro rate of 20-hydroxyecdysone metabolization, those clones known to be resistant to the steroid were 'high metabolizers' (> 70% relative rate), whereas the sensitive clones were 'poor metabolizers' (< 30% relative rate). Hence, it is tempting to conclude that ecdysteroid resistance of the Chironomus cell clones is due to metabolic inactivation of the steroid hormone. PMID- 9342222 TI - Palytoxin-induced channel formation within the Na+/K+-ATPase does not require a catalytically active enzyme. AB - It has been demonstrated that palytoxin binds to and forms a channel within the Na+/K+-ATPase. To investigate whether palytoxin-induced channel formation within the sodium pump can occur independently of ATP hydrolysis and phosphorylation of the enzyme, an Asp369-->Ala mutant of the alpha1 subunit of the sheep sodium pump was produced and coexpressed with beta subunits in the yeast Saccharomyces cerevisiae. This aspartic acid residue, which during ion transport becomes phosphorylated from ATP, is essential for the function of the sodium pump. Therefore, as expected, microsomes isolated from yeast expressing the mutant sodium pump do not exhibit any ouabain sensitive ATPase activity, whereas in microsomes from yeast expressing the wild-type sodium pump, 60% of the total ATPase activity is ouabain-sensitive. Ouabain binds to yeast membranes containing either wild-type or mutant sodium pumps with similar Bmax (1.45+/-0.05 versus 1.37+/-0.02 pmol/mg) and Kd values (27.7+/-0.91 versus 29.57+/-0.93 nM), thus indicating that the mutant sodium pumps are expressed in the yeast and that the mutation does not considerably affect the conformation of the enzyme. In the presence of phosphate ouabain binds to microsomes containing the wild-type sodium pump with a Kd of 3.62+/-0.34 nM, showing that, although not necessary, phosphoenzyme formation enhances binding of the steroid. Phosphate or ATP, however, inhibit binding of ouabain to microsomes containing the mutant sodium pump with IC50 values of 78+/-3 microM and 3.0+/-0.4 microM, respectively. Despite these radical changes in the interactions of the mutant enzyme with ouabain, the interactions with palytoxin are not affected by the mutation. Palytoxin causes K+ efflux from yeast cells expressing the wild-type or mutant sodium pumps with EC50 values of 3.5+/-0.4 nM and 6.2+/-0.9 nM, respectively. Palytoxin-induced efflux from cells expressing wild-type or mutant sodium pumps occurs with similar t1/2 values of 20.3+/-2.1 min and 22.2+/-3.1 min, respectively. Ouabain inhibits K+ efflux from both cell types with IC50 values of 28+/-2 microM and 210+/-15 microM, respectively. Cells expressing the Asp369- >Ala mutants have an IC50 7.5-fold higher than that obtained with cells expressing the wild-type sodium pumps, possibly because ATP or phosphate present in the cytosol of the yeast cells influence and decrease ouabain binding to the mutant sodium pump. Thus, while ouabain binding and the associated inhibition of ion fluxes is promoted by phosphorylation of the wild-type enzyme by phosphate or ATP, palytoxin-induced channel formation is independent of phosphorylation and can be separated from the ATPase function of the sodium pump. Since ion fluxes through the sodium pump protein do not depend on ATP hydrolysis, the results suggest that the ionophores of pumps and ion channels might share common structural features. PMID- 9342223 TI - Chloroplast SecA functions as a membrane-associated component of the Sec-like protein translocase of pea chloroplasts. AB - Protein cross-linking studies with a thylakoid membrane translocation intermediate were used to demonstrate that chloroplast SecA functions as a membrane-associated component of the Sec-like ATP-dependent protein translocase of pea chloroplasts. In assays with isolated thylakoids, it was observed that translocation of the 33-kDa protein of the oxygen-evolving complex of photosystem II (OE33) decreased when the ATP concentration was low, and that the protein accumulated as a bound precursor. The bound precursor was able to be translocated into the lumen when the ATP concentration was raised, indicating that the precursor was bound to the translocation apparatus. Inclusion of apyrase in the import reaction prevented translocation but did not affect precursor binding to the membrane. When this translocation intermediate was treated with the cross linking agent disuccinimidyl suberate, a single predominant cross-linked product of 120 kDa was produced. This conjugate could be immunoprecipitated with antibodies to pea chloroplast SecA, identifying the cross-linking partner as SecA. This provides direct evidence for a functional interaction between a thylakoid precursor protein and a component of the thylakoid protein translocation apparatus. PMID- 9342224 TI - Reduced turnover of the D1 polypeptide and photoactivation of electron transfer in novel herbicide resistant mutants of Synechocystis sp. PCC 6803. AB - Two missense mutants, A263P and S264P, and a deletion mutant des-Ala263, Ser264, have been constructed in the D1 protein of the cyanobacterium Synechocystis sp PCC 6803. All were expected to induce a significant conformational change in the QB-binding region of photosystem II (PSII). Although the des-Ala263, Ser264-D1 mutant accumulated some D1 protein in the thylakoid membrane it was unable to grow photoautotrophically or evolve oxygen. Thermoluminescence and chlorophyll fluorescence studies confirmed that this deletion mutant did not show any functional PSII activity. In contrast, [S264P]D1 was able to grow photoautotrophically and give light-saturated rates of oxygen evolution at 60% of the rate of the wild-type control strain, TC31. The A263P missense mutant was also able to evolve oxygen at 50% of TC31 rates although it did not readily grow photoautotrophically. Thermoluminescence, flash oxygen yield and chlorophyll fluorescence measurements indicated that in both missense mutants electron transfer from QA to QB was significantly impaired in dark adapted cells. However, QA to QB electron transfer could be photoactivated in the mutants by background illumination. Both the A263P and S264P mutants also showed an increase in resistance to the s-triazine family of herbicides although this feature did not hold for the phenolic herbicide, ioxynil. Of particular interest was that the two missense mutants, especially S264P, possessed a slower rate of turnover of the D1 protein compared with TC31 and in vivo contained detectable levels of a 41-kDa adduct consisting of D1 and the alpha subunit of cytochrome b559. When protein synthesis was blocked by the addition of lincomycin, D1 degradation was again slower in S264P than TC31. The results are discussed in terms of structural changes in the QB-binding region which affect herbicide and plastoquinone binding and perturb the normal regulatory factors that control the degradation of the D1 protein and its synchronisation with the synthesis of a replacement D1 protein. PMID- 9342225 TI - Undersulfation of cartilage proteoglycans ex vivo and increased contribution of amino acid sulfur to sulfation in vitro in McAlister dysplasia/atelosteogenesis type 2. AB - Mutations in the diastrophic dysplasia sulfate transporter gene cause a family of chondrodysplasias including, in order of increasing severity, diastrophic dysplasia, atelosteogenesis type 2 and achondrogenesis type 1B. McAlister dysplasia is a lethal chondrodysplasia considered on the basis of minor radiographic features to be a disorder different from atelosteogenesis type 2. Here, we demonstrate that McAlister dysplasia arises from mutations in the diastrophic dysplasia sulfate transporter gene and that this disorder essentially coincides on molecular and biochemical grounds with atelosteogenesis type 2. The fetus affected by McAlister dysplasia we have studied is a compound heterozygote for mutations leading to R279W and N425D substitutions in the diastrophic dysplasia sulfate transporter. Proteoglycan sulfation was studied in epiphyseal cartilage and in chondrocyte cultures of the patient by high performance liquid chromatography of chondrotinase digested proteoglycans; a high amount of non sulfated disaccharide was observed as a consequence of the alteration of the transporter function caused by the mutations. However, sulfated disaccharides were detectable even if in low amounts, both in cultured cells and tissue. Functional impairment of the sulfate transporter was demonstrated in vitro by reduced incorporation of [35S]sulfate relative to [3H]glucosamine in proteoglycans synthesized by chondrocytes and by sulfate-uptake assays in fibroblasts. Parallel in vitro studies in a patient with achondrogenesis 1B indicated that the severity of the clinical phenotype seems to be correlated to the residual activity of the sulfate transporter. The capacity of fibroblasts to use cysteine as an alternative source of sulfate was evaluated by double-labeling experiments. Relative incorporation of [35S]cysteine-derived sulfate in the glycosaminoglycan chains was increased in the patient's cells, indicating that, in vitro, the catabolism of sulfur-containing amino acids can partially compensate for intracellular sulfate deficiency. Residual sulfation observed in proteoglycans extracted from cartilage suggests that this mechanism may be operating also in vivo. PMID- 9342227 TI - The effects of high salt concentrations on the regulation of the substrate specificity of human recombinant deoxycytidine kinase. AB - Deoxycytidine kinase (dCK) is a salvage pathway enzyme with broad substrate specificity that can phosphorylate both pyrimidine and purine deoxynucleosides, including important antiviral and cytostatic agents. The kinetic behaviour of dCK is complex with saturation curves showing negative cooperativity. In this study, we have expressed and affinity purified recombinant dCK, using the pET 9d vector system with a histidine tag-sequence and a thrombin cleavage site fused to the N terminus of the dCK coding sequence. The His-tagged protein showed essentially the same kinetic properties as the protease cleaved protein and the purified protein isolated from human spleen. However, addition of 0.2-0.4 M NaCl during the dCK reaction caused a stimulation of 2'-deoxycytidine (dCyd), and the antileukemic analog 2-chlorodeoxyadenosine (CldAdo) phosphorylation, but an inhibition of the 2'-deoxyguanosine (dGuo) phosphorylation, both with His-tagged and protease cleaved dCK. The negative cooperativity observed with dCyd was eliminated by the presence of 0.4 M NaCl so that the Hill coefficient changed from 0.6 to 1.4. In contrast, dGuo phosphorylation that initially followed Michaelis-Menten kinetics showed negative cooperativity after addition of 0.4 NaCl. The alterations in kinetic properties were not accompanied by any apparent changes in subunit structure as revealed by gel filtration. The major form of dCK eluted in the position corresponding to a dimer in the presence or absence of salt, but a minor fraction of dCK, eluting in the position of a tetramer, was diminished in the presence of salt. The mechanism for the effects of 0.4 M NaCl on dCK kinetic behaviour is not known but it is most likely due to alterations in the conformation of the active site of the enzyme. The effects described here also may explain some of the discrepancies reported in the literature on the substrate specificity of this complex enzyme. PMID- 9342226 TI - Cloning and characterization of Manduca sexta and Plutella xylostella midgut aminopeptidase N enzymes related to Bacillus thuringiensis toxin-binding proteins. AB - We report the purification, cloning and characterization of an aminopeptidase N from the midgut epithelium of Manduca sexta that binds Cry1Ab5, an insecticidal crystal protein [ICP] from Bacillus thuringiensis. Sequence information derived from this M. sexta aminopeptidase N was used for the cloning of an aminopeptidase N from the midgut brush-border membrane of Plutella xylostella, an insect species of which some populations acquired resistance against Cry1Ab5. Affinity chromatography on a Cry1Ab5 matrix was used to isolate a 120-kDa glycoprotein from the larval midgut of the lepidopteran M. sexta. On ligand blots the purified 120-kDa protein discriminates between the lepidopteran-specific Cry1Ab5 and the coleopteran-specific Cry3A delta-endotoxin. Internal amino acid sequences from the 120-kDa protein were used for the design of degenerate oligonucleotides. From a nested PCR with M. sexta midgut cDNA as template, a DNA fragment was obtained which shows similarity to prokaryotic and eukaryotic aminopeptidase N genes. This PCR fragment was used to screen cDNA libraries of larval midguts from M. sexta and P. xylostella. From the M. sexta midgut cDNA library a 2973-bp nucleotide sequence was cloned. The ORF of the sequence encodes a 942-residue aminopeptidase N (M. sexta Apn2) containing two hydrophobic regions. The NH2-terminal hydrophobic region corresponds to a secretory signal sequence and the COOH terminal hydrophobic region is typical of glycosylphosphatidylinositol (glycosyl PtdIns)-anchored proteins. Low-stringency hybridization of the P. xylostella midgut cDNA library with M. sexta apn2 probes enabled the isolation of a 3118-bp sequence with an ORF encoding a 946-residue preproprotein. This aminopeptidase N (P. xylostella Apn1) displays 61% amino acid identity to M. sexta Apn2 and contains a COOH-terminal signal peptide for glycosyl-PtdIns anchor addition. Both M. sexta Apn2 and P. xylostella Apn1 contain four Cys residues, which are highly conserved among eukaryotic aminopeptidase N molecules. Treatment of Sf9 cells expressing the P. xylostella apn1 gene with PtdIns-specific phospholipase C demonstrated that P. xylostella Apn1 is attached to the insect cell membrane by a glycosyl-PtdIns anchor. PMID- 9342229 TI - Type-1 plasminogen-activator inhibitor -- conformational differences between latent, active, reactive-centre-cleaved and plasminogen-activator-complexed forms, as probed by proteolytic susceptibility. AB - We have analysed the susceptibility of latent, active, reactive-centre-cleaved and plasminogen-activator-complexed type-1 plasminogen-activator inhibitor (PAI 1) to the non-target proteinases trypsin, endoproteinase Asp-N, proteinase K and subtilisin. This analysis has allowed us to detect conformational differences between the different forms of PAI-1 outside the reactive-centre loop and beta sheet A. Proteinase-hypersensitive sites were clustered in three regions. Firstly, susceptibility was observed in the region around alpha-helix E, beta strand 1A, and the flanking loops, which are believed to form flexible joints during movements of beta-sheet A. Secondly, hypersensitive sites were observed in the loop between alpha-helix I and beta-strand 5A. Thirdly, the gate region, encompassing beta-strands 3C and 4C, was highly susceptible to trypsin in latent PAI-1, but not in the other conformations. The digestion patterns differed among all four forms of PAI-1, indicating that each represents a unique conformation. The differential proteolytic susceptibility of the flexible-joint region may be coupled to the differential affinity to vitronectin, binding in the same region. The analysis also allowed detection of conformational differences between reactive-centre-cleaved forms produced under different solvent conditions. The digestion pattern of plasminogen-activator-complexed PAI-1 was different from that of active PAI-1, but indistinguishable from that of one of the reactive centre-cleaved forms, as the complexed and this particular cleaved PAI-1 were completely resistant to all the non-target proteinases tested. This observation is in agreement with the notion that complex formation involves reactive-centre cleavage and a large degree of insertion of the reactive-centre loop into beta sheet A. Our analysis has allowed the identification of some flexible regions that appear to be implicated in the conformational changes during the movements of beta-sheet A and during the inhibitory reaction of serpins with their target proteinases. PMID- 9342228 TI - Biosynthesis of the proteoglycan decorin -- identification of intermediates in galactosaminoglycan assembly. AB - Biosynthesis of decorin was investigated by incubating a rat fibroblast cell line with various radiolabelled protein and galactosaminoglycan precursors. The following cell-associated and distinct intermediates were isolated and identified: a pool of non-glycosylated core protein, two pools of decorin with incomplete chains, one with three sulphated disaccharide repeats and another with five or more sulphated disaccharide repeats, as well as decorin with mature chains. Results of pulse/chase experiments indicated that these pools represented discrete stages in chain growth. Treatment with brefeldin A, which blocks transport from the endoplasmic reticulum to the Golgi, resulted in accumulation of decorin with an incomplete chain containing six or seven largely unsulphated disaccharide repeats. During recovery from drug treatment, 4-sulfation reappeared earlier than 6-sulfation. The results suggest that the galactosaminoglycan assembly-line consists of separate multienzyme complexes that build only a limited section of the chain. Furthermore, brefeldin A causes segregation of compartments involved in separate stages of the assembly line. In an earlier report [Moses, J., Oldberg. A., Cheng, F. & Fransson, L.-A. (1997) Eur. J. Biochem. 248, 521-526] we took advantage of such segregation to identify and characterize a transient 2-phosphorylation of xylose in the linkage region. PMID- 9342230 TI - Tissue-specific and stage-specific expression of two silkworm ecdysone receptor isoforms -- ecdysteroid-dependent transcription in cultured anterior silk glands. AB - Previously, we isolated a cDNA clone for the ecdysone receptor B1 isoform of the silkworm, Bombyx mori (BmEcR-B1). Here we report the cloning of a cDNA that encodes the Bombyx ecdysone receptor A isoform (BmEcR-A) and mRNA expression of the two BmEcR isoforms during molting and metamorphosis. At larval-pupal transformation, mRNA expression of BmEcR-B1 was predominant in most tissues examined, including three larval tissues (midgut, epidermis, and fat body) and the wing imaginal disc. The anterior silk gland was the only tissue where BmEcR-A was predominant. These expression patterns were different from observations demonstrated in Drosophila. In the anterior silk gland, both EcR isoforms were expressed synchronously during the fifth larval instar, while expression of the A isoform preceded that of the B1 isoform by two days in the fourth instar. Precedence of BmEcR-A during the fourth instar and synchronization of both isoforms during the fifth instar were also observed in the middle and posterior silk glands, suggesting that transcription of BmEcR in the silk gland is regulated differently in these two instars. In the cultured anterior silk glands of day 0 of the fifth instar, transcription of BmEcR-A and BmEcR-B1 was induced dose dependently by more than 5 ng/ml 20-hydroxyecdysone. BmEcR-A and BmEcR-B1 mRNAs were induced within 2 h and 1 h, respectively, of the addition of 20 hydroxyecdysone. These results suggest that the increase of BmEcR mRNAs during the fifth instar is induced in vivo by a small increase in ecdysteroids. PMID- 9342231 TI - The stathmin family -- molecular and biological characterization of novel mammalian proteins expressed in the nervous system. AB - Stathmin is a ubiquitous phosphoprotein proposed to be a relay integrating various intracellular signaling pathways. Its high phylogenetic conservation and the identification of the related molecules, SCG10 in rat and XB3 in Xenopus, suggested the existence of a stathmin-related family. A systematic PCR-based approach allowed the identification of several novel mammalian sequences of which two coded for expressed members of the stathmin family; the translated RB3 sequence shares 88% amino-acid identity with that of XB3 and is thus its rat homologue, and RB3' corresponds to an alternatively spliced product of the same gene, encoding a truncated form. Within their stathmin-like domain, the alpha helix, probably responsible for coiled-coil protein-protein interactions, is conserved, as well as are two consensus phosphorylation sites; in their N terminal extension domain, two cystein residues most likely responsible for membrane attachment through palmitoylation, are present in RB3/RB3' as in SCG10. The novel identification and characterization of the corresponding proteins showed that all three are associated with the particulate, membrane-containing fraction. They furthermore display several spots of decreasing pI on two dimensional immunoblots, suggesting that they are phosphorylated in vivo. As for SCG10, RB3 mRNA is detectable only in the nervous system by in situ hybridization, but at similar levels in the newborn and the adult brain as revealed by Northern blots, whereas SCG10 expression decreases in the adult. Furthermore, RB3 mRNA is undetectable in PC12 cells, whereas SCG10 mRNA increases after treatment with nerve growth factor, inducing neuronal differentiation. In conclusion, we demonstrate here the existence of a highly conserved stathmin related family in mammals, of which each member seems to play specific roles, related to the control of cell proliferation and activities for stathmin and to that of neuronal differentiation for SCG10, the novel RB3/RB3' proteins being rather related to the expression of differentiated neuronal functions. PMID- 9342232 TI - Molecular cloning of the gene for mouse leukotriene-C4 synthase. AB - Leukotriene C4 (LTC4) synthase (LTC4S), an integral membrane protein, catalyzes the conjugation of leukotriene A4 with reduced glutathione to form LTC4, the biosynthetic parent of the additional cysteinyl leukotriene metabolites. An XmnI digested fragment of a P1 clone from a 129 mouse ES library contained the full length gene of 2.01 kb for mouse LTC4S. The mouse LTC4S gene is comprised of 5 exons of 122, 100, 71, 82 and 241 nucleotides, with intron sizes that range from 76 nucleotides to 937 nucleotides. The intron/exon boundaries are identical to those of the human genes for LTC4S and 5-lipoxygenase-activating protein (FLAP). Primer extension demonstrated a single transcription-initiation site 64 bp 5' of the ATG translation-start site. Nucleotide sequencing of 1.2 kb of the 5' flanking region revealed multiple putative sites for activating protein-2, CCAAT/enhancer-binding protein, and polyoma virus enhancer-3. Fluorescent in situ hybridization mapped the mouse LTC4S gene to mouse chromosome 11, in a region containing the genes for interleukin 13 and granulocyte/macrophage-colony stimulating factor, and orthologous to the chromosomal location of 5q35 for the human LTC4S gene. Thus, the mouse LTC4S gene is similar in size, intron/exon organization and chromosomal localization to the human LTC4S gene. Recent mutagenic analysis of the conjugation function of human LTC4S has identified R51 and Y93 as critical for acid and base catalysis of LTA4 and reduced glutathione, respectively. A comparison across species for proteins that possess LTC4S activity reveals conservation of both of these residues, whereas R51 is absent in the FLAP molecules. Thus, within the glutathione S-transferase superfamily of genes, alignment of specific residues allows the separation of LTC4S family members from their most structurally similar counterparts, the FLAP molecules. PMID- 9342233 TI - Purification of the cardiac sarcoplasmic reticulum membrane protein phospholamban from recombinant Escherichia coli. AB - Phospholamban (PLN) was expressed in Escherichia coli as a protein fusion with glutathione S-transferase (GST). GST-PLN was mostly present in the insoluble protein fraction and accounted for approximately 50% of total insoluble protein. Attempts to suppress inclusion body formation or to use GST as an affinity purification tag failed. A successful purification method is based on preparative SDS/PAGE and electrodialysis. From 1 g cells we typically purified 13.5 mg fusion protein with a PLN content of 2.8 mg. We genetically inserted an enterokinase (EK) protease site just in front of the PLN sequence and demonstrated the proteolytical liberation of PLN from the carrier protein. The approach described represents a substantial advancement in PLN expression and purification. PMID- 9342234 TI - The catalytic subunit of Dictyostelium cAMP-dependent protein kinase -- role of the N-terminal domain and of the C-terminal residues in catalytic activity and stability. AB - The C subunit of Dictyostelium cAMP-dependent protein kinase (PKA) is unusually large (73 kDa) due to the presence of 330 amino acids N-terminal to the conserved catalytic core. The sequence following the core, including a C-terminal -Phe-Xaa Xaa-Phe-COOH motif, is highly conserved. We have characterized the catalytic activity and stability of C subunits mutated in sequences outside the catalytic core and we have analyzed their ability to interact with the R subunit and with the heat-stable protein-kinase inhibitor PKI. Mutants carrying deletions in the N terminal domain displayed little difference in their kinetic properties and retained their capacity to be inhibited by R subunit and by PKI. In contrast, the mutation of one or both of the phenylalanine residues in the C-terminal motif resulted in a decrease of catalytic activity and stability of the proteins. Inhibition by the R subunit or by PKI were however unaffected. Sequence comparison analysis of other protein kinases revealed that a -Phe-Xaa-Xaa-Phe- motif is present in many Ser/Thr protein kinases, although its location at the very end of the polypeptide is a particular feature of the PKA family. We propose that the presence of this motif may serve to identify isoforms of protein kinases. PMID- 9342235 TI - Relocation of Syk protein-tyrosine kinase to the actin filament network and subsequent association with Fak. AB - Previous studies demonstrated that Syk protein-tyrosine kinase (Syk) is activated by thrombin in platelets. To elucidate the function of Syk in platelets, we have biochemically examined the intracellular location of Syk and the molecules associated with Syk, following platelet activation. In human platelets, thrombin induces the relocation of Syk to the cytoskeletal fraction presumably via Syk tyrosine phosphorylation. Relocated Syk is associated with the actin filament network, and the early phase (10-90 s) of this association can be partially inhibited by the pretreatment of platelets with cytochalasin D, an inhibitor of actin polymerization. Upon thrombin stimulation, Syk becomes associated with Fak as demonstrated by co-immunoprecipitation. The association of both kinases can be inhibited by pretreatment of platelets with cytochalasin D. Interestingly, reconstitution experiments, using COS cells transfected with various porcine Syk mutants, revealed that the kinase domain, but not the kinase activity, of Syk is required for the association of Syk with the actin filament network. These findings suggest that thrombin-induced association of Syk with Fak correlates with the state of actin polymerization, and may play an important role in platelet activation. PMID- 9342237 TI - A putative beta-tubulin phosphate-binding motif is involved in lateral microtubule protofilament interactions. AB - We have investigated the role of a putative GTP-binding beta-tubulin motif in microtubule polymerization. A peptide containing residues 126-142 of the beta tubulin subunit (peptide G) was synthesised and an antibody against it raised. Peptide G prevents the binding of GTP to tubulin and also microtubule polymerization but not the formation of vinblastine-induced tubulin spirals, suggesting that it may prevent lateral but not longitudinal tubulin-tubulin interactions. The antibody to peptide G shows little reaction with the interphase microtubule network, mitotic spindles or midbody of cultured cells, whereas it clearly reacts with vinblastine-induced paracrystals. These results suggest that this putative phosphate-binding site present in beta-tubulin could be involved in the lateral tubulin-tubulin interactions along the microtubule structure. PMID- 9342236 TI - Characterization of gelsolin truncates that inhibit actin depolymerization by severing activity of gelsolin and cofilin. AB - Gelsolin is a calcium-activated actin-binding protein with six subdomains. The N terminal (G1) domain is essential for actin-filament-severing activity while other domains within G2-3 position the protein on the filament side allowing G1 to sever. In order to generate reagents capable of competitively inhibiting endogenous gelsolin and, potentially, other actin filament regulatory protein, we expressed several truncates of gelsolin in Escherichia coli, and analyzed how they affected the in vitro activity of two different actin-binding proteins, gelsolin and cofilin. A Ca2+-sensitive truncate containing G2-6 inhibited the F actin-depolymerizing activities of both gelsolin and cofilin, while a G2-3 truncate was less effective. Using two independent assays, our results support the idea that gelsolin truncates inhibit actin filament severing and do not markedly affect actin subunit dissociation kinetics. Cosedimentation assays in the presence of calcium demonstrate that the G2-6 truncate binds to F-actin more strongly than the G2-3 truncate consistent with a protection mechanism by conformational change of F-actin and/or competitive binding to actin filaments which depends upon the presence of actin filament binding domains. PMID- 9342238 TI - The 70-kDa heat-shock protein/DnaK chaperone system is required for the productive folding of ribulose-biphosphate carboxylase subunits in Escherichia coli. AB - We have studied the in vivo requirements of the DnaK chaperone system for the folding of recombinant ribulose-bisphosphate carboxylase/oxygenase in Escherichia coli. Expression of functional dimeric or hexadecameric ribulose-bisphosphate carboxylase from different bacterial sources (including purple bacteria and cyanobacteria) was severely impaired in E. coli dnaK, dnaJ, or grpE mutants. These enzymes were synthesized mostly in soluble, fully enzymatically active forms in wild-type E. coli cells cultured in the temperature range 20-42 degrees C, but aggregated extensively in dnaK null mutants. Co-expression of dnaK, but not groESL, markedly reduced the aggregation of ribulose-bisphosphate carboxylase subunits in dnaK null mutants and restored the enzyme activity to levels found in isogenic wild-type strains. Ribulose-bisphosphate carboxylase expression in wild type E. coli cells growing at 30 degrees C promoted an enhanced synthesis of stress proteins, apparently by sequestering DnaK from its negative regulatory role in this response. The overall results indicate that the DnaK chaperone system assists in vivo the folding pathway of ribulose-bisphosphate carboxylase large subunits, most probably at its very early stages. PMID- 9342239 TI - Cloning of two putative ecdysteroid receptor isoforms from Tenebrio molitor and their developmental expression in the epidermis during metamorphosis. AB - Using the Drosophila EcR-B1 cDNA as a probe, we have cloned the putative ecdysteroid receptor from the mealworm Tenebrio molitor. We have isolated two cDNAs with different 5' termini that contain a complete open reading frame. These two cDNAs encode two proteins with distinct N-terminal regions corresponding to two isoforms. The coleopteran receptor is obviously related to the ecdysteroid receptor of other insects, but shares only 89% and 61% amino acid identities with the DNA-binding and ligand-binding domains of the Drosophila receptor, respectively. Its expression pattern has been examined in the epidermis during the last larval instar and pupal stage of T. molitor, in correlation with the hemolymph ecdysteroid titer. Hybridizations revealed two transcripts of 7 kb and 6.5 kb detected in most stages during metamorphosis and corresponding to the A and B1 isoforms. These two mRNAs are highly evident just before the rise of each ecdysteroid peak both in prepupae and in pupae. They show almost the same expression pattern in epidermis except for the second part of the pupal stage, during which only the A isoform is detected. PMID- 9342240 TI - Characterization of the embryonic globin chains of the marsupial Tammar wallaby, Macropus eugenii. AB - The embryonic hemoglobins of the marsupial Tammar wallaby (Macropus eugenii) are known to aggregate, which was shown by the finding that the Hill coefficient, h, was greater than 4.0 in the upper part of the oxygen equilibrium curve. Here, we have undertaken a detailed primary structure analysis of the Tammar wallaby pouch young hemoglobin complement, which we hoped might provide clues into the residues that cause aggregation and a high embryonic h. The Tammar wallaby embryonic hemoglobin complement is principally four major hemoglobins each with a different isoelectric point. Two early expressed hemoglobins contain the same embryonic beta-like chain, epsilon (epsilon), but two separate alpha-like chains, termed zeta and zeta prime (zeta and zeta') both of which are N-terminally blocked. The later two expressed hemoglobins contain the same adult alpha-chain, but different beta-like chains. The latest expressed hemoglobin contains the same beta-like chain, epsilon, as the two early expressed forms, but the third expressed hemoglobin contains a unique beta-like chain which we have termed omega (omega). A protein database similarity search using the first 54 N-terminal amino acids of the omega-chain showed a range of sequence identities of 57-72% to all known mammalian beta-like chains, including the other marsupial epsilon-chains. The closest identity, reflected by both the highest percentage identity and Smith Waterman score, was with the embryonic beta-chains of the aves. While the primary structures of the hemoglobins reported here do not explain the low hemoglobin oxygen affinity in embryonic marsupial blood, the finding of the similarity with the bird globin-like sequence with one of the marsupial chains has implications on mammalian globin evolution. How many other marsupials and placental mammals are harboring a bird-like globin in their embryos? PMID- 9342241 TI - Two-dimensional 1H-NMR and CD structural analysis in a micellar medium of a bovine alphaS1-casein fragment having benzodiazepine-like properties. AB - The conformation of the benzodiazepine-like decapeptide, YLGYLEQLLR, corresponding to residues 91-100 of bovine alphaS1-casein, has been examined in SDS micelles using CD, two-dimensional 1H-NMR and restrained molecular-dynamics simulation. Evidence is presented that the decapeptide adopts a rigid structure in water/SDS micellar medium, but not in water or dimethylsulfoxide. The three dimensional structure, consistent with the proton-proton distances obtained from the quantitative analysis of the two-dimensional NOEs, was generated by restrained energy minimization and molecular-dynamics simulation. In water/SDS micellar medium, YLGYLEQLLR adopts an amphipathic helicoid structure with distinct hydrophobic and hydrophilic faces. The relative disposition of the tyrosine aromatic rings was compared with that of the aromatic rings in the benzodiazepines. PMID- 9342242 TI - Evidence for a cysteine-histidine thioether bridge in functional units of molluscan haemocyanins and location of the disulfide bridges in functional units d and g of the betaC-haemocyanin of Helix pomatia. AB - In functional units d and g from the betaC-haemocyanin of the gastropod Helix pomatia, aminoacid analysis in the presence of dimethyl sulfoxide showed the occurrence of seven cysteine residues. Titration with 5,5'-dithiobis(2 nitrobenzoic acid), however, did not reveal any free thiol group. Pepsinolysis at pH 2.0 followed by amino acid analysis and partial sequencing of the cysteine containing peptides showed that six cysteine residues are involved in the formation of three disulfide bridges at corresponding positions. The results indicated that the remaining cysteine residue is linked by a thioether bridge to a histidine residue located two positions further in the sequence (H. pomatia d Cys60 His62; H. pomatia g Cys66 His68). This residue corresponds to one of the three histidine residues considered to be involved in the coordination of the copper A atom of the dinuclear copper group of the functional units. The presence of the thioether bond was further evidenced by an absorption band at 255 nm and by molecular mass determinations with electrospray mass spectrometry on a peptic peptide from H. pomatia d and on peptides obtained by proteolysis of carboxymethylated H. pomatia d with trypsin and pronase. Upon sequence analysis the cysteine-histidine thioether bridge was cleaved into didehydroalanine (polymers) and 2-thiolhistidine. A peptide with a cysteine-histidine thioether bridge was isolated from pepsinolysates of functional units c, e, f, g and h of Sepia officinalis and unit g of Octopus vulgaris, obtained from haemocyanin of the cephalopods S. officinalis and O. vulgaris. A cysteine-histidine thioether bridge, which was reported previously for tyrosinase of Neurospora crassa, thus seems to be a general feature for functional units of molluscan haemocyanins. PMID- 9342243 TI - Purification and characterization of an alcohol:N,N-dimethyl-4-nitrosoaniline oxidoreductase from the methanogen Methanosarcina barkeri DSM 804 strain Fusaro. AB - Cell-free extracts of Methanosarcina barkeri DSM 804 showed alcohol dehydrogenase activity under aerobic conditions when N,N-dimethyl-4-nitrosoaniline (NDMA) was used as an artificial electron acceptor. The NDMA-dependent alcohol dehydrogenase (NDMA-ADH) was purified to approximate homogeneity by column chromatography. It is most probably a homodimeric enzyme consisting of subunits of 45 kDa, the native molecular mass estimated by gel filtration being about 87 kDa. The purified protein had an isoelectric point of 4.3. It possesses a tightly but noncovalently bound NADP(H) cofactor. Each subunit contains 1 mol NADP(H)/mol, about 2 mol Zn2+/mol and significant amounts of magnesium. The purified enzyme preferably oxidized primary alcohols (including benzyl alcohol). NDMA-ADH from M. barkeri also catalyzed the stoichiometric dismutation of aldehydes, especially higher aliphatic aldehydes, to form equimolar amounts of the corresponding alcohol and acid without addition of an electron carrier. The enzyme did not catalyze the dehydrogenation of methanol or the disproportionation of formaldehyde and therefore is not directly involved in methanogenesis. An alignment of the N-terminal amino acid sequence of the enzyme with the sequences of other alcohol dehydrogenases from methanogenic and nonmethanogenic bacteria indicated no significant identity. Nevertheless there was a quite interesting sequence similarity in the first 30 N-terminal amino acids to plant cinnamyl alcohol dehydrogenase. NDMA-ADH from M. barkeri is a novel type of alcohol dehydrogenase in methanogenic bacteria. PMID- 9342244 TI - Pro108 is important for folding and stabilization of adrenal ferredoxin, but does not influence the functional properties of the protein. AB - The truncated mutant Met-adrenodoxin-(4-107)-peptide of bovine adrenal ferredoxin was expressed as apoprotein in Escherichia coli BL21 and could be reconstituted to the holoform by chemical or enzymatic methods. The reconstituted protein had spectroscopic, functional and redox properties similar to the Met-adrenodoxin-(4 108)-peptide of adrenal ferredoxin, into which the cluster was inserted upon expression in the same Escherichia coli strain. Rate of in vitro cluster insertion into the Met-adrenodoxin-(4-107) apoprotein was much lower than for the Met-adrenodoxin-(4-108) apoprotein under identical conditions. Comparative thermodynamic studies with the Met-adrenodoxin-(4-108)-peptide indicated that removal of Pro108 resulted in an extensive decrease of the overall stability of the protein in either oxidation state. The Met-adrenodoxin-(4-107)-peptide showed a higher sensitivity to urea denaturation and had a sensibly lower denaturation temperature, 44.8 degrees C, compared with 51.7 degrees C for mutant Met adrenodoxin-(4-108). The stability of the reduced state of both mutants is slightly lower than that of the oxidized state indicating that this protein region does not undergo major structural changes upon reduction. PMID- 9342245 TI - The Kw recombinase, an integrase from Kluyveromyces waltii. AB - Site-specific recombinases of the integrase family share limited amino-acid sequence similarity, but use a common reaction mechanism to recombine distinct DNA target sites. Here we report the characterisation of the Kw site-specific recombinase, encoded on the 2 mu-like plasmid pKWS1 from the yeast Kluyveromyces waltii. Using in vitro-translated Kw recombinase, we show that the protein is able to bind and to recombine its putative DNA target site. Recombination is conservative and the Kw target site has a spacer of seven base pairs. We show that Kw recombinase is able to mediate recombination in a mammalian cell line, thus, it has potential for use as a tool for genomic manipulation in heterologous systems. PMID- 9342246 TI - Catalytic characteristics of tryparedoxin. AB - Tryparedoxin, a thioredoxin-related protein from Crithidia fasciculata with a molecular mass of 16 kDa catalyses the reduction of a peroxiredoxin-type peroxidase, Cf21, at the expense of trypanothione [Nogoceke, E., Gommel, D. U., Kiess, M., Kalisz, H. M. & Flohe, L. E. (1997) Biol. Chem. Hoppe-Seyler 378, 827 836]. The kinetic analysis of tryparedoxin revealed an enzyme substitution mechanism. The corresponding molecular event was elucidated to be a reversible oxidoreduction of the disulfide bridge in the thioredoxin-related motif WCPPC. The amino-proximal cysteine residue of this active site was more reactive in S alkylation experiments than the distal residue. The natural substrates of tryparedoxin, trypanothione and Cf21, could only be substituted by glutathione and glutathione disulfide with considerable loss in activity. The pronounced specificity of tryparedoxin is further accentuated by low limiting Km values for Cf21 and trypanothione (2.2 microM and 130 microM, respectively, as compared to 990 microM for gluthathione disulfide and an infinite value for glutathione). Tryparedoxin can therefore be classified as a trypanothione: peroxiredoxin oxidoreductase. The reduction of tryparedoxin by trypanothione appears to be the rate-limiting step in the trypanothione-dependent hydroperoxide reduction because(a) the regeneration of reduced tryparedoxin from the tryparedoxin trypanothione complex is rate limiting (k[cat] 392 min[-1]), (b) the physiological trypanothione concentrations may not always saturate tryparedoxin, and (c) the rate constants for the net forward reaction of Cf21 are faster than those of the tryparedoxin reaction. The functional characteristics of tryparedoxin explain the limited capacity of trypanosomatids in coping with oxidative stress and qualify the enzyme as a potential target for the design of specific trypanocidal compounds. PMID- 9342247 TI - The active species of 'CO2' utilized by formylmethanofuran dehydrogenase from methanogenic Archaea. AB - Formylmethanofuran dehydrogenase from methanogenic Archaea catalyzes the reversible conversion of CO2 and methanofuran to formylmethanofuran, which is an intermediate in methanogenesis from CO2, a biological process yielding approximately 0.3 billion tons of CH4 per year. With the enzyme from Methanosarcina barkeri, it is shown that CO2 rather than HCO3- is the active species of 'CO2' utilized by the dehydrogenase. Evidence is also presented that the enzyme catalyzes a methanofuran-dependent exchange between CO2 and the formyl group of formylmethanofuran. The results are consistent with N carboxymethanofuran being an intermediate in CO2 reduction to formylmethanofuran. PMID- 9342248 TI - Inactivation kinetics of the reduced spinach chloroplast fructose-1,6 bisphosphatase by subtilisin. AB - The course of inactivation of the reduced spinach chloroplast fructose-1,6 bisphosphatase by digestion with subtilisin has been followed by the progress curve method [Tsou, C. L. (1988) Adv. Enzymol. 61, 381-436] and found to follow first-order kinetics. On the basis of the hydrolysis of the substrate, fructose 1,6-bisphosphate, at different concentrations during proteolysis by subtilisin, the first-order inactivation rate constants for the free enzyme and the enzyme substrate complex can both be determined. The ratio between the inactivation rate constants for the free enzyme and the enzyme-substrate complex indicates strong protection against subtilisin proteolysis by the substrate. It is proposed that the above ratio can be used as a quantitative measure of substrate protection for enzyme inactivation generally. As it has been found that the site of proteolysis is located in a loop region near the N-terminus and well away from the active site, the substrate protection indicates a conformation change of the enzyme away from the substrate binding site. PMID- 9342249 TI - Study of the substrate-binding properties of bovine liver adenosine kinase and inhibition by fluorescent nucleoside analogues. AB - Adenosine kinase (AK) catalyzes the phosphorylation of adenosine to AMP with ATP as phosphate donor. Intrinsic fluorescence of bovine liver AK was shown previously to be a sensitive probe to quantify the binding of substrates to the enzyme [Elaloui, A., Divita, G., Maury, G., Imbach, J.-L. & Goody, R. S. (1994) Eur. J Biochem. 221, 839-846]. AK contains two catalytic, sites: a high-affinity site, which binds adenosine and AMP selectively; and a site for ATP and ADP. In the present work, these two sites were characterized by combining the quenching of protein fluorescence induced by the binding of the ligands and the fluorescence enhancement observed upon binding of the N-methylanthraniloyl derivated nucleotides or adenosine. A new fluorescent analog of adenosine, 5'-N methylanthraniloyl-adenosine, was synthesized and shown to bind selectively to the high-affinity adenosine-binding site with an affinity similar to that of adenosine (Kd 1 microM). In contrast, 2'(3')-N-methylanthraniloyl derivatives of ATP, adenosine (5')tetraphospho(5')adenosine (Ap4A), and adenosine (5')pentaphospho(5')adenosine (Ap5A), bind to the enzyme at the ATP site. Methylantraniloyl derivatives of ATP and adenosine were used as tools for selective characterization of a series of adenosine analogues. The bisubstrate inhibitors Ap4A and Ap5A bind to the ATP site with high affinity and apparently not to the adenosine site, thus acting more as ATP analogues than true bisubstrate ligands. The binding properties of a series of adenosine analogues were strongly dependent on the structural modifications on adenosine. The analogues modified at positions 2' or 3' show similar affinities for AK as that of adenosine, whereas adenosine analogues modified at the base present a relatively low affinity for the enzyme. PMID- 9342251 TI - Activation of latent HIV-1 by Mycobacterium tuberculosis and its purified protein derivative in alveolar macrophages from HIV-infected individuals in vitro. AB - Although Mycobacterium tuberculosis (MTB) and its purified protein derivative (PPD) induce HIV in cell lines that harbor latent HIV infection, it is not known whether similar activation of HIV in primary macrophages infected with HIV occurs. This possibility was examined using alveolar macrophages (AM) obtained by bronchoalveolar lavage of HIV-infected subjects with CD4 counts <200/microl. PPD induced transcription of HIV in AM from HIV-infected subjects by reverse transcription-polymerase chain reaction (RT-PCR). PPD and MTB infection also induced HIV production in AM from these HIV-infected patients, determined by HIV p24 enzyme-linked immunosorbent assay (ELISA). Viral production in AM required short periods of cell contact with allogeneic lymphocytes. HIV was only inducible, however, in AM from subjects with detectable HIV load (one to three copies of HIV DNA/1000 cells). Thus, MTB and its PPD can induce HIV in latently infected AM. PMID- 9342250 TI - Structural aspects of the interaction of peptidyl-glycylleucine-carboxyamide, a highly potent antimicrobial peptide from frog skin, with lipids. AB - The interaction of PGLa (peptidyl-glycylleucine-carboxyamide), a 21-amino-acid residue cationic peptide, isolated from the skin of the South African clawed frog, Xenopus laevis, with model membrane systems was investigated. Our studies focussed on the importance of the difference in the phospholipid composition of bacterial and erythrocyte membranes. This is of particular interest to gain information on the specificity of membranolysis exhibited by this peptide against bacteria but not against erythrocytes. In phosphate buffer at physiological pH, as well as in the presence of the zwitterionic phosphatidylcholine and sphingomyelin. the peptide had a random structure but it adopted an alpha-helical conformation in the presence of negatively charged lipids. Furthermore, calorimetric experiments showed that PGLa had no effects on the thermotropic phase behavior of liposomes composed of the choline phosphatides, while separation of a distinct peptide-rich domain was observed for phosphatidylglycerol liposomes. In addition to the main transition of pure 1,2 dipalmitoylglycerophosphoglycerol at 40 degrees C a second transition owing to the peptide-perturbed lipid domains was found at 41 degrees C. This conclusion is supported by X-ray diffraction experiments which indicated that PGLa penetrates into the hydrophobic core of the bilayer inducing an untilting of the hydrocarbon chains as observed in the gel phase of the pure lipid. These results demonstrate that this antibacterial peptide specifically interacts with negatively charged lipid membranes, which are characteristic of bacterial membranes. This can be explained based on the structural features of PGLa. PMID- 9342252 TI - Progressive encephalopathy associated with CD4/CD8 inversion in adult FIV infected cats. AB - Experimental intravenous challenge of five adult cats with the feline immunodeficiency virus Maryland isolate (FIV-MD) was investigated for its ability to induce neurologic abnormalities associated with the onset of immunodeficiency. Five 8-month-old cats were inoculated with 1000 median tissue culture infective dose of FIV-MD isolate, with five age-matched cats serving as uninfected controls. All FIV-MD-infected cats tested positive for serum antiviral antibodies and plasma viral DNA as detected by polymerase chain reaction at 2, 4, 10, and 16 months postinfection (PI). At 10 and 16 months PI, there was a significant reduction in the CD4/CD8 lymphocyte ratio, with all cats having a CD4/CD8 ratio of 1 or less. Total protein electrophoretic analysis of cerebrospinal fluid demonstrated a significantly increased albumin quotient at 4 and 16 months PI, representing a disrupted blood-brain barrier (BBB). At 16 months PI, all cats demonstrated a preferential increase in frontal cortical slow-wave activity compared with control cats. Serial evaluation of brainstem auditory evoked potential recordings revealed a prolongation of the interpeak latencies times over the study time. At least one abnormality was found over time in visual and somatosensory evoked potential testing in three and four infected cats, respectively. Comparing lymphocyte subtype ratios with neurologic testing revealed that every FIV-MD-infected cat exhibited an abnormality in at least one neurologic functional test with a concurrent CD4/CD8 count ratio of 1 or less. Overall, this study demonstrated that FIV-MD infection in adult cats results in a delayed-onset, progressive encephalopathy that parallels the decline in the CD4/CD8 lymphocyte ratio. Compared with prior information from pediatric FIV-MD infected cats, these results indicate that age of infection influences the onset and severity of disease and may be associated with CD4 cell depletion in FIV-MD infected cats, as seen in HIV-1-infected humans. PMID- 9342253 TI - Clinical manifestations and predictors of survival in older women infected with HIV. AB - To review the natural history of HIV infection in older women, a retrospective review of women enrolled in the HIV Outpatient Program based at the Medical Center of Louisiana in New Orleans was performed. Eighty-four of the women were at least 40 years of age. Older women were more likely to be diagnosed with selected psychosocial illnesses (e.g., injection drug use, alcohol abuse, anxiety, depression, psychosis, dementia) compared with women <40 years of age. There was no association with age and other opportunistic processes or HIV related symptoms, but cervical dysplasia and chlamydia cervicitis were less common in older women. In a multivariate proportional hazards model, characteristics predictive for death among older women included a CD4 cell count <200 cells/mm3 (relative risk [RR], 2.86; 95% confidence interval [CI], 1.18, 6.86; p < .02), a diagnosis of an opportunistic process (RR, 3.25; 95% CI, 1.24, 8.55; p < .02), antiretroviral combination therapy (RR, 0.36; 95% CI, 0.12, 1.13: p < .08), and hormone replacement therapy (HRT) (RR, 0.28; 95% CI, 0.07, 1.10; p < .06). HRT should be considered in the management of postmenopausal HIV-infected women for its known documented benefits shown in populations of persons not infected with HIV. Prospective studies to better evaluate risks and benefits of HRT in HIV-infected women are warranted. PMID- 9342254 TI - Predictors of survival in HIV-infected persons with 50 or fewer CD4 cells/mm3. AB - The objective of this study was to identify prognostic factors for survival in patients with pretreatment CD4 < or =50 cells/mm3 treated with nucleoside analogs, and to develop and validate a mortality risk model based on these factors. The design of the study consisted of retrospective analysis of AIDS Clinical Trials Group (ACTG) protocols 116a, 116b/117, 155, and 118. The setting was the multicenter AIDS Clinical Trials Group. The patients were HIV-infected with pretreatment CD4 < or =50 cells/mm3 and various degrees of prior zidovudine (ZDV) use. Double-blind, three-arm randomized control trials ACTG 116a and ACTG 116b/117 compared ZDV with didanosine (ddI). ACTG 155 compared ZDV with zalcitabine or combination therapy. Our validation study, ACTG 118, compared the effects of three different doses of ddI on survival. The main outcome measures were survival and mortality. The three studies combined enrolled 699 patients with entry CD4 T-lymphocyte counts of < or =50 cells/mm3. Forty percent of patients died during follow-up, with a median survival of 19.7 months. Multivariate analysis showed shorter survival at p < 0.0001 with lower CD4 count (relative hazard [RH] = 0.98) and lower hemoglobin level (RH = 0.81). Other factors included older age (RH = 1.03), male gender (RH = 1.70), Hispanic ethnicity (RH = 1.68), and symptomatic disease stage (RH = 2.06). Our predictive mortality risk model differentiated well patients with differing risks of mortality. When the risk model was applied to ACTG 118, the validation data set, the identified prognostic factors could distinguish patients with varying risks of death (p < 0.001, stratified log-rank test). These results demonstrate that CD4 T-lymphocytes counts < or =50 cells/mm3 should not be considered a precursor of imminent death; considerable variability in survival exists in severely immunocompromised patients. Our identification of prognostic indicators for survival can aid clinicians and patients in management of their disease and researchers in design of future clinical trials. PMID- 9342255 TI - Application of an expert system in the management of HIV-infected patients. AB - A rule-based expert system, Customized Treatment Strategies for HIV (CTSHIV), which encodes information from the literature on known drug-resistant mutations was developed. Additional rules include ranking and weighting based on antiviral activities, redundant mechanisms of action, overlapping toxicities, relative levels of drug-resistance, and proportion of drug-resistant clones in the HIV quasispecies. Plasma was obtained from HIV-infected patients and the RNA was extracted. Segments of the HIV pol gene encoding the entire protease, reverse transcriptase, and integrase proteins were amplified by reverse transcriptase polymerase chain reaction (using a total of three primer pairs) and cloned. Sequencing was performed on five clones from each of two patients. When the patient's RNA sequencing data were entered into the expert program, and the information was downloaded directly into the CTSHIV program, the five most effective two, three, and four drug regimens coupled with an explanation for their choice were displayed for each patient. Thus, the CTSHIV system couples efficient genetic sequencing with an expert program that recommends regimens based on information in the current medical literature. It may serve as a useful tool in the design of clinical trials and in the management of HIV-infected patients. PMID- 9342256 TI - Survival after AIDS diagnosis in a cohort of hemophilia patients. Multicenter Hemophilia Cohort Study. AB - We studied factors affecting survival after the diagnosis of AIDS in a cohort of 1253 patients with hemophilia. The nature of the AIDS-defining condition was found to be as important as age at seroconversion and CD4+ lymphocyte level in predicting survival. A multivariate analysis yielded estimates of median survival for groups defined by age at seroconversion (0 through 15, 16 through 69), CD4+ lymphocyte count (<100 cells/microl versus > or = 100 cells/microl), and 10 AIDS defining disease groups. Estimates of median survival after a single AIDS defining condition ranged from 3 to 51 months, depending on the diseases. Median survival after a second AIDS-defining condition was about 1.5- to 2.0-fold shorter than after an initial, isolated AIDS-defining condition. HIV-related neurologic disease (i.e., AIDS dementia complex or multifocal leukoencephalopathy) was a notable exception. It correlated with the shortest estimates of median survival (3 to 9 months), and this poor prognosis was no worse for patients who had a second AIDS-defining condition. The results of this analysis were consistent in most respects with other published analyses of factors affecting survival. These findings may be useful in the clinical care of persons with AIDS and in estimating the number of persons alive who have had a particular AIDS-defining disease. PMID- 9342257 TI - High risk of HIV-related mortality is associated with selenium deficiency. AB - To determine the independent contribution of specific immunologic and nutritional factors on survival in HIV-1 disease, CD4 cell count, antiretroviral treatment, plasma levels of vitamins A, E, B6, and B12 and minerals selenium and zinc were considered in relation to relative risk for HIV-related mortality. Immune parameters and nutrients known to affect immune function were evaluated at 6 month intervals in 125 HIV-1-seropositive drug-using men and women in Miami, FL, over 3.5 years. A total of 21 of the HIV-1-infected participants died of HIV related causes during the 3.5-year longitudinal study. Subclinical malnutrition (i.e., overly low levels of prealbumin, relative risk [RR] = 4.01, p < 0.007), deficiency of vitamin A (RR = 3.23, p < 0.03), vitamin B12 deficiency (RR = 8.33, p < 0.009), zinc deficiency (RR = 2.29.1, p < 0.04), and selenium deficiency (RR = 19.9, p < 0.0001) over time, but not zidovudine treatment, were shown to each be associated with HIV-1-related mortality independent of CD4 cell counts <200/mm3 at baseline, and CD4 counts over time. When all factors that could affect survival, including CD4 counts <200/mm3 at baseline, CD4 levels over time, and nutrient deficiencies were considered jointly, only CD4 counts over time (RR = 0.69, p < 0.04) and selenium deficiency (RR = 10.8, p < 0.002) were significantly associated with mortality. These results indicate that selenium deficiency is an independent predictor of survival for those with HIV-1 infection. PMID- 9342258 TI - HIV-1 risk and vaccine acceptability in the Ugandan military. AB - Between July and October 1993, 570 19- to 22-year-old volunteers were screened for HIV-1, with a resulting seroprevalence rate of 18.3% (95% CI: 14.0%, 22.6%). A cohort of 249 HIV-1-noninfected military recruits in the Ugandan Peoples' Defense Forces was followed prospectively for up to 18 months to document rates of HIV-1 seroprevalence, seroconversion, and knowledge and attitudes related to vaccine acceptability. The HIV-1 seroincidence rate was 3.56 per 100 person-years (95% CI: 1.49, 5.62) over 309 person-years of observation. At the 3- and 12-month visits, subjects were interviewed on issues of acceptance and knowledge about vaccines, including anti-HIV vaccines in particular. More than 90% believe that HIV vaccines will not cause HIV infection, and if offered, 88% report that they would take the vaccine if they were not already infected. Nonvaccine prevention methods were considered less reliable; monogamy and condom use were considered effective by only 33.5% and 69.3% of the cohort respectively. After completing the vaccine acceptability questionnaire at the 12-month visit, subjects were offered an approved polyvalent meningococcal vaccine as an indicator of general vaccine acceptance. All subjects reported receiving at least one previous vaccination, and 95% willingly accepted the meningococcal vaccination. The Ugandan military is a stable population at substantial risk for HIV-1 infection and may be a suitable population for vaccine efficacy trials. PMID- 9342259 TI - Prevalence and risk factors for HTLV-II infection in 913 injecting drug users in Stockholm, 1994. AB - The prevalence and risk factors for acquisition of human T-cell lymphotropic virus type I and II (HTLV-I and II) were investigated in a prospective study of 913 injecting drug users (IDUs) in Stockholm in 1994. Epidemiologic data were recorded, and blood samples were tested for antibodies against HTLV-I and HTLV II; human immunodeficiency virus (HIV) types 1 and 2; and hepatitis A (HAV), B (HBV), C (HCV), and D (HDV). Positive serologic results for HTLV were confirmed by Western blot (WB) and polymerase chain reaction (PCR). Of the 905 participants with conclusive HTLV-II status, 29 (3.2%) were HTLV-II positive, and all but three were of Nordic descent. None was HTLV-I infected. One person was infected as early as 1981, before HIV had reached the IDU population in Sweden. The prevalence of HTLV-II infection was 12% among HIV-1-seropositive and 1.8% among HIV-1-seronegative participants. The overall seroprevalences were 14% for HIV-1, 0% for HIV-2, 41% for HAV, 75% for HBV, 92% for HCV, and 8% for HDV. Although amphetamine has been the main injecting drug in Sweden for several decades, heroin abuse combined with a debut of injecting drugs before 1975 was identified as the most important risk factor associated with HTLV-II infection. HAV and HIV seropositivity were also independent risk factors. PMID- 9342260 TI - Decreased survival of HTLV-I carriers in leprosy patients from the Democratic Republic of the Congo: a historical prospective study. AB - In this historical prospective study using sera stored for 22 years, we investigated the effect of HTLV-I infection on survival in a population of leprosy patients in the Democratic Republic of the Congo (formerly Zaire). We also determined the distribution of HTLV-I by subpopulation, age, and gender. Stored sera taken from a population of leprosy patients and controls in 1969 were tested for HTLV-I. Follow-up survival data on these patients were obtained in 1991. The sera collected in 1969 from 520 individuals was used to determine the prevalence of HTLV-I. Included in this number were 328 patients resident in the sanatorium. Survival and other data were available for 327 of these. A multivariate survival analysis using a logistic regression model was performed to evaluate the influence of HTLV-I status, age, type of leprosy, gender, duration of hospitalization, and ethnic group on survival. The overall prevalence of HTLV I among the 520 individuals in the prevalence study was 34%, with 37.4% in the leprosy group and 25.2% in the control group (p < 0.01). Multivariate analysis using logistic regression showed that females of the Mongo and Ngombe ethnic group taken together were significantly more likely to be infected than the other groups (OR = 3.67, 95% CI: 2.14 to 6.30). A comparison of the death rates directly standardized for age and sex showed that the rate was significantly higher for HTLV-I positive (5.5/100 person-years of observation) compared with HTLV-I negative (3.6/100 person-years of observation). A survival analysis using the Cox model showed a risk ratio of 1.4 (CI: 1.04 to 1.89) for those infected with HTLV-I. An increase in the death rate was associated with HTLV-I infection in leprosy inpatients. The decreased survival associated with HTLV-I infection may result from an increased susceptibility to a variety of diseases. PMID- 9342261 TI - Modified amplicor HIV-1 polymerase chain reaction assay in Thailand. PMID- 9342262 TI - Thalidomide use is associated with weight gain in HIV-1-positive clients. PMID- 9342304 TI - Functional bioinformatics: the cellular response database. AB - Biological Scientists function in an increasingly data rich environment. The emerging field of bioinformatics is attempting to insure that this flow of information can be structured to support the generation of significant biological hypothesis and ultimately new knowledge. To date, most of the current databases have focused on protein and nucleic acid sequence information as the principle type of data stored for further interpretation. In this paper, we describe the Cellular Response Database. This database stores functional information regarding the changes of cellular gene expression associated with various stimuli, and supports queries linking cell types, expressed genes, and inducers. The database is designed to support information-intensive queries to aid in the determination of biological function, and is flexible enough to allow the storage of a broad range of experimental data such as cytotoxicity data, immunoassays of target gene protein expression, and others. PMID- 9342263 TI - HIV-1 encodes a sequence overlapping env gp41 with highly significant similarity to selenium-dependent glutathione peroxidases. PMID- 9342305 TI - Laccase and melanin in the pathogenesis of Cryptococcus neoformans. AB - Cryptococcosis, caused by an encapsulated fungus, Cryptococcus neoformans, has emerged as a life threatening infection in HIV positive individuals and other immunocompromised hosts. The present review describes laccase and its product melanin as an important virulence factor of Cryptococcus neoformans and illustrates the approaches used in elucidating the pathogenesis of cryptococcosis. Characterization of the biochemical pathways leading to melanin synthesis is summarized using biochemical and biomolecular approaches. Melanin synthesis is dependent on a single copper-dependent enzyme, laccase. Since the mammalian host does not contain this enzyme, laccase is an attractive candidate for the study of fungal pathogenesis, as well as a drug target. The cloning of the CNLAC1 gene and construction of CNLAC1 gene knock-out strains has confirmed its role in the virulence of Cryptococcus. Also described is the role of melanin in the host-pathogen interactions. Melanin may protect Cryptococcus cells by a variety of methods including anti-oxidant or cell wall surface effects thereby offering protection against numerous effectors of cellular immunity. PMID- 9342306 TI - Fluorescence correlation spectroscopy: diagnostics for sparse molecules. AB - The robust glow of molecular fluorescence renders even sparse molecules detectable and susceptible to analysis for concentration, mobility, chemistry, and photophysics. Correlation spectroscopy, a statistical-physics-based tool, gleans quantitative information from the spontaneously fluctuating fluorescence signals obtained from small molecular ensembles. This analytical power is available for studying molecules present at minuscule concentrations in liquid solutions (less than one nanomolar), or even on the surfaces of living cells at less than one macromolecule per square micrometer. Indeed, routines are becoming common to detect, locate, and examine individual molecules under favorable conditions. PMID- 9342307 TI - IkappaB kinase: beginning, not the end. PMID- 9342308 TI - Once there were twenty. PMID- 9342309 TI - Recombinational crossroads: eukaryotic enzymes and the limits of bacterial precedents. PMID- 9342310 TI - Phosphotyrosine signaling and the single cell:metazoan boundary. PMID- 9342311 TI - Twist and shout (and pull): molecular chiropractors undo DNA. PMID- 9342312 TI - On roads not taken in the evolution of protein catalysts: antibody steroid isomerases that use an enamine mechanism. AB - Reactive immunization has emerged as a new tool for the study of biological catalysis. A powerful application resulted in catalytic antibodies that use an enamine mechanism akin to that used by the class I aldolases. With regard to the evolution of enzyme mechanisms, we investigated the utility of an enamine pathway for the allylic rearrangement exemplified by Delta5-3-ketosteroid isomerase (KSI; EC 5.3.3.1). Our aldolase antibodies were found to catalyze the isomerization of both steroid model compounds and steroids. The kinetic and chemical studies showed that the antibodies afforded rate accelerations up to a factor of 10(4) by means of an enamine mechanism in which imine formation was the rate-determining step. In light of our observations and the enzyme studies by other workers, we suggest that an enamine pathway could have been an early, viable KSI mechanism. Although this pathway is amenable to optimization for increased catalytic power, it appears that certain factors precluded its evolution in known KSI enzymes. PMID- 9342313 TI - Making chemistry selectable by linking it to infectivity. AB - The link between recognition and replication is fundamental to the operation of the immune system. In recent years, modeling this process in a format of phage display combinatorial libraries has afforded a powerful tool for obtaining valuable antibodies. However, the ability to readily select and isolate rare catalysts would expand the scope of library technology. A technique in which phage infection controlled the link between recognition and replication was applied to show that chemistry is a selectable process. An antibody that operated by covalent catalysis to form an acyl intermediate restored phage infectivity and allowed selection from a library in which the catalyst constituted 1 in 10(5) members. Three different selection approaches were examined for their convenience and generality. Incorporating these protocols together with well known affinity labels and mechanism-based inactivators should allow the procurement of a wide range of novel catalytic antibodies. PMID- 9342315 TI - Protein-protein interactions among the Aux/IAA proteins. AB - The plant hormone indoleacetic acid (IAA) transcriptionally activates early genes in plants. The Aux/IAA family of early genes encodes proteins that are short lived and nuclear-localized. They also contain a putative prokaryotic betaalphaalpha DNA binding motif whose formation requires protein dimerization. Here, we show that the pea PS-IAA4 and Arabidopsis IAA1 and IAA2 proteins perform homo- and heterotypic interactions in yeast using the two-hybrid system. Gel filtration chromatography and chemical cross-linking experiments demonstrate that the PS-IAA4 and IAA1 proteins interact to form homodimers in vitro. Deletion analysis of PS-IAA4 indicates that the betaalphaalpha containing acidic C terminus of the protein is necessary for homotypic interactions in the yeast two hybrid system. Screening an Arabidopsis lambda-ACT cDNA library using IAA1 as a bait reveals heterotypic interactions of IAA1 with known and newly discovered members of the Arabidopsis Aux/IAA gene family. The new member IAA24 has similarity to ARF1, a transcription factor that binds to an auxin response element. Combinatorial interactions among the various members of the Aux/IAA gene family may regulate a variety of late genes as well as serve as autoregulators of early auxin-regulated gene expression. These interactions provide a molecular basis for the developmental and tissue-specific manner of auxin action. PMID- 9342316 TI - Characterization of recombinant phytochrome from the cyanobacterium Synechocystis. AB - The complete sequence of the Synechocystis chromosome has revealed a phytochrome like sequence that yielded an authentic phytochrome when overexpressed in Escherichia coli. In this paper we describe this recombinant Synechocystis phytochrome in more detail. Islands of strong similarity to plant phytochromes were found throughout the cyanobacterial sequence whereas C-terminal homologies identify it as a likely sensory histidine kinase, a family to which plant phytochromes are related. An approximately 300 residue portion that is important for plant phytochrome function is missing from the Synechocystis sequence, immediately in front of the putative kinase region. The recombinant apoprotein is soluble and can easily be purified to homogeneity by affinity chromatography. Phycocyanobilin and similar tetrapyrroles are covalently attached within seconds, an autocatalytic process followed by slow conformational changes culminating in red-absorbing phytochrome formation. Spectral absorbance characteristics are remarkably similar to those of plant phytochromes, although the conformation of the chromophore is likely to be more helical in the Synechocystis phytochrome. According to size-exclusion chromatography the native recombinant apoproteins and holoproteins elute predominantly as 115- and 170-kDa species, respectively. Both tend to form dimers in vitro and aggregate under low salt conditions. Nevertheless, the purity and solubility of the recombinant gene product make it a most attractive model for molecular studies of phytochrome, including x-ray crystallography. PMID- 9342317 TI - Prp43: An RNA helicase-like factor involved in spliceosome disassembly. AB - The Saccharomyces cerevisiae genes PRP2, PRP16, and PRP22 encode pre-mRNA splicing factors that belong to the highly conserved "DEAH" family of putative RNA helicases. We previously identified two additional members of this family, JA1 and JA2. To investigate its biological function, we cloned the JA1 gene and generated alleles carrying mutations identical to those found in highly conserved regions of other members of the DEAH family. A ja1 allele carrying a mutation identical to that in the temperature-sensitive (ts) prp22-1 gene conferred ts phenotype when integrated into the genome of a wild-type strain by gene replacement. Northern analysis of RNA obtained from the ts strain shifted to a nonpermissive temperature revealed accumulation of unspliced pre-mRNAs and excised intron lariats. Furthermore, analysis of splicing complexes showed that intron lariats accumulated in spliceosomes. The results presented indicate that JA1 encodes a pre-mRNA processing factor (Prp) involved in disassembly of spliceosomes after the release of mature mRNA. We have therefore renamed this gene PRP43. PMID- 9342318 TI - Cloning of mDEAH9, a putative RNA helicase and mammalian homologue of Saccharomyces cerevisiae splicing factor Prp43. AB - Yeast splicing factor Prp43, a DEAH box protein of the putative RNA helicase/RNA dependent NTPase family, is a splicing factor that functions late in the pre-mRNA splicing pathway to facilitate spliceosome disassembly. In this paper we report cDNA cloning and characterization of mDEAH9, an apparent mammalian homologue of Prp43. Amino acid sequence comparison revealed that the two proteins are approximately 65% identical over a 500-aa region spanning the central helicase domain and the C-terminal region. Expression of mDEAH9 in S. cerevisiae bearing a temperature-sensitive mutation in prp43 was sufficient to restore growth at the nonpermissive temperature. This functional complementation was specific, as mouse mDEAH9 failed to complement mutations in related splicing factor genes prp16 or prp22. Finally, double label immunofluorescence experiments performed with mammalian cells revealed colocalization of mDEAH9 and splicing factor SC35 in punctate nuclear speckles. Thus, the hypothesis that mDEAH9 represents the mammalian homologue of yeast Prp43 is supported by its high sequence homology, functional complementation, and colocalization with a known splicing factor in the nucleus. Our results provide additional support for the hypothesis that the spliceosomal machinery that mediates regulated, dynamic changes in conformation of pre-mRNA and snRNP RNAs has been highly conserved through evolution. PMID- 9342319 TI - Structural evidence for a second sialic acid binding site in avian influenza virus neuraminidases. AB - The x-ray structure of a complex of sialic acid (Neu5Ac) with neuraminidase N9 subtype from A/tern/Australia/G70C/75 influenza virus at 4 degrees C has revealed the location of a second Neu5Ac binding site on the surface of the enzyme. At 18 degrees C, only the enzyme active site contains bound Neu5Ac. Neu5Ac binds in the second site in the chair conformation in a similar way to which it binds to hemagglutinin. The residues that interact with Neu5Ac at this second site are mostly conserved in avian strains, but not in human and swine strains, indicating that it has some as-yet-unknown biological function in birds. PMID- 9342320 TI - The crystal structure of human interferon beta at 2.2-A resolution. AB - Type I interferons (IFNs) are helical cytokines that have diverse biological activities despite the fact that they appear to interact with the same receptor system. To achieve a better understanding of the structural basis for the different activities of alpha and beta IFNs, we have determined the crystal structure of glycosylated human IFN-beta at 2.2-A resolution by molecular replacement. The molecule adopts a fold similar to that of the previously determined structures of murine IFN-beta and human IFN-alpha2b but displays several distinct structural features. Like human IFN-alpha2b, human IFN-beta contains a zinc-binding site at the interface of the two molecules in the asymmetric unit, raising the question of functional relevance for IFN-beta dimers. However, unlike the human IFN-alpha2b dimer, in which homologous surfaces form the interface, human IFN-beta dimerizes with contact surfaces from opposite sides of the molecule. The relevance of the structure to the effects of point mutations in IFN-beta at specific exposed residues is discussed. A potential role of ligand-ligand interactions in the conformational assembly of IFN receptor components is discussed. PMID- 9342321 TI - Glu-tRNAGln amidotransferase: a novel heterotrimeric enzyme required for correct decoding of glutamine codons during translation. AB - The three genes, gatC, gatA, and gatB, which constitute the transcriptional unit of the Bacillus subtilis glutamyl-tRNAGln amidotransferase have been cloned. Expression of this transcriptional unit results in the production of a heterotrimeric protein that has been purified to homogeneity. The enzyme furnishes a means for formation of correctly charged Gln-tRNAGln through the transamidation of misacylated Glu-tRNAGln, functionally replacing the lack of glutaminyl-tRNA synthetase activity in Gram-positive eubacteria, cyanobacteria, Archaea, and organelles. Disruption of this operon is lethal. This demonstrates that transamidation is the only pathway to Gln-tRNAGln in B. subtilis and that glutamyl-tRNAGln amidotransferase is a novel and essential component of the translational apparatus. PMID- 9342323 TI - Synthesis and screening of small molecule libraries active in binding to DNA. AB - Five synthetic combinatorial libraries of 2,080 components each were screened as mixtures for inhibition of DNA binding to two transcription factors. Rapid, solution-phase synthesis coupled to a gel-shift assay led to the identification of two compounds active at a 5- to 10-microM concentration level. The likely mode of inhibition is intercalation between DNA base pairs. The efficient deconvolution through sublibrary synthesis augurs well for the use of large mixtures of small, nonpeptide molecules in biological screens. PMID- 9342322 TI - Cloning of the cDNA for the TATA-binding protein-associated factorII170 subunit of transcription factor B-TFIID reveals homology to global transcription regulators in yeast and Drosophila. AB - The human transcription factor B-TFIID is comprised of TATA-binding protein (TBP) in complex with one TBP-associated factor (TAF) of 170 kDa. We report the isolation of the cDNA for TAFII170. By cofractionation and coprecipitation experiments, we show that the protein encoded by the cDNA encodes the TAF subunit of B-TFIID. Recombinant TAFII170 has (d)ATPase activity. Inspection of its primary structure reveals a striking homology with genes of other organisms, yeast MOT1, and Drosophila moira, which belongs to the Trithorax group. Both homologs were isolated in genetic screens as global regulators of pol II transcription. This supports our classification of B-TFIID as a pol II transcription factor and suggests that specific TBP-TAF complexes perform distinct functions during development. PMID- 9342324 TI - The amino-terminal region of Tyk2 sustains the level of interferon alpha receptor 1, a component of the interferon alpha/beta receptor. AB - Tyk2 belongs to the Janus kinase (JAK) family of receptor associated tyrosine kinases, characterized by a large N-terminal region, a kinase-like domain and a tyrosine kinase domain. It was previously shown that Tyk2 contributes to interferon-alpha (IFN-alpha) signaling not only catalytically, but also as an essential intracellular component of the receptor complex, being required for high affinity binding of IFN-alpha. For this function the tyrosine kinase domain was found to be dispensable. Here, it is shown that mutant cells lacking Tyk2 have significantly reduced IFN-alpha receptor 1 (IFNAR1) protein level, whereas the mRNA level is unaltered. Expression of the N-terminal region of Tyk2 in these cells reconstituted wild-type IFNAR1 level, but did not restore the binding activity of the receptor. Studies of mutant Tyk2 forms deleted at the N terminus indicated that the integrity of the N-terminal region is required to sustain IFNAR1. These studies also showed that the N-terminal region does not directly modulate the basal autophosphorylation activity of Tyk2, but it is required for efficient in vitro IFNAR1 phosphorylation and for rendering the enzyme activatable by IFN-alpha. Overall, these results indicate that distinct Tyk2 domains provide different functions to the receptor complex: the N-terminal region sustains IFNAR1 level, whereas the kinase-like domain provides a function toward high affinity ligand binding. PMID- 9342325 TI - Structure of the thrombin complex with triabin, a lipocalin-like exosite-binding inhibitor derived from a triatomine bug. AB - Triabin, a 142-residue protein from the saliva of the blood-sucking triatomine bug Triatoma pallidipennis, is a potent and selective thrombin inhibitor. Its stoichiometric complex with bovine alpha-thrombin was crystallized, and its crystal structure was solved by Patterson search methods and refined at 2.6-A resolution to an R value of 0.184. The analysis revealed that triabin is a compact one-domain molecule essentially consisting of an eight-stranded beta barrel. The eight strands A to H are arranged in the order A-C-B-D-E-F-G-H, with the first four strands exhibiting a hitherto unobserved up-up-down-down topology. Except for the B-C inversion, the triabin fold exhibits the regular up-and-down topology of lipocalins. In contrast to the typical ligand-binding lipocalins, however, the triabin barrel encloses a hydrophobic core intersected by a unique salt-bridge cluster. Triabin interacts with thrombin exclusively via its fibrinogen-recognition exosite. Surprisingly, most of the interface interactions are hydrophobic. A prominent exception represents thrombin's Arg-77A side chain, which extends into a hydrophobic triabin pocket forming partially buried salt bridges with Glu-128 and Asp-135 of the inhibitor. The fully accessible active site of thrombin in this complex is in agreement with its retained hydrolytic activity toward small chromogenic substrates. Impairment of thrombin's fibrinogen converting activity or of its thrombomodulin-mediated protein C activation capacity upon triabin binding is explained by usage of overlapping interaction sites of fibrinogen, thrombomodulin, and triabin on thrombin. These data demonstrate that triabin inhibits thrombin via a novel and unique mechanism that might be of interest in the context of potential therapeutic applications. PMID- 9342326 TI - Characterization of a genetically engineered inactivation-resistant coagulation factor VIIIa. AB - Individuals with hemophilia A require frequent infusion of preparations of coagulation factor VIII. The activity of factor VIII (FVIII) as a cofactor for factor IXa in the coagulation cascade is limited by its instability after activation by thrombin. Activation of FVIII occurs through proteolytic cleavage and generates an unstable FVIII heterotrimer that is subject to rapid dissociation of its subunits. In addition, further proteolytic cleavage by thrombin, factor Xa, factor IXa, and activated protein C can lead to inactivation. We have engineered and characterized a FVIII protein, IR8, that has enhanced in vitro stability of FVIII activity due to resistance to subunit dissociation and proteolytic inactivation. FVIII was genetically engineered by deletion of residues 794-1689 so that the A2 domain is covalently attached to the light chain. Missense mutations at thrombin and activated protein C inactivation cleavage sites provided resistance to proteolysis, resulting in a single-chain protein that has maximal activity after a single cleavage after arginine-372. The specific activity of partially purified protein produced in transfected COS-1 monkey cells was 5-fold higher than wild-type (WT) FVIII. Whereas WT FVIII was inactivated by thrombin after 10 min in vitro, IR8 still retained 38% of peak activity after 4 hr. Whereas binding of IR8 to von Willebrand factor (vWF) was reduced 10-fold compared with WT FVIII, in the presence of an anti-light chain antibody, ESH8, binding of IR8 to vWF increased 5-fold. These results demonstrate that residues 1690-2332 of FVIII are sufficient to support high-affinity vWF binding. Whereas ESH8 inhibited WT factor VIII activity, IR8 retained its activity in the presence of ESH8. We propose that resistance to A2 subunit dissociation abrogates inhibition by the ESH8 antibody. The stable FVIIIa described here provides the opportunity to study the activated form of this critical coagulation factor and demonstrates that proteins can be improved by rationale design through genetic engineering technology. PMID- 9342327 TI - Respiratory chain is required to maintain oxidized states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli cells. AB - DsbA, the disulfide bond catalyst of Escherichia coli, is a periplasmic protein having a thioredoxin-like Cys-30-Xaa-Xaa-Cys-33 motif. The Cys-30-Cys-33 disulfide is donated to a pair of cysteines on the target proteins. Although DsbA, having high oxidizing potential, is prone to reduction, it is maintained essentially all oxidized in vivo. DsbB, an integral membrane protein having two pairs of essential cysteines, reoxidizes DsbA that has been reduced upon functioning. It is not known, however, what might provide the overall oxidizing power to the DsbA-DsbB disulfide bond formation system. We now report that E. coli mutants defective in the hemA gene or in the ubiA-menA genes markedly accumulate the reduced form of DsbA during growth under the conditions of protoheme deprivation as well as ubiquinone/menaquinone deprivation. Disulfide bond formation of beta-lactamase was impaired under these conditions. Intracellular state of DsbB was found to be affected by deprivation of quinones, such that it accumulates first as a reduced form and then as a form of a disulfide-linked complex with DsbA. This is followed by reduction of the bulk of DsbA molecules. These results suggest that the respiratory electron transfer chain participates in the oxidation of DsbA, by acting primarily on DsbB. It is remarkable that a cellular catalyst of protein folding is connected to the respiratory chain. PMID- 9342328 TI - RecA tests homology at both pairing and strand exchange. AB - RecA is a 38-kDa protein from Escherichia coli that polymerizes on single stranded DNA, forming a nucleoprotein filament that pairs with homologous duplex DNA and carries out strand exchange in vitro. To observe the effects of mismatches on the kinetics of the RecA-catalyzed recombination reaction, we used assays based upon fluorescence energy transfer that can differentiate between the pairing and strand displacement phases. Oligonucleotide sequences that produced 2 14% mismatches in the heteroduplex product of strand exchange were tested, as well as completely homologous and heterologous sequences. The equilibrium constant for pairing decreased as the number of mismatches increased, which appeared to result from both a decrease in the rate of formation and an increase in the rate of dissociation of the intermediates. In addition, the rate of strand displacement decreased with increasing numbers of mismatches, roughly in proportion to the number of mismatches. The equilibrium constant for pairing and the rate constant for strand displacement both decreased 6-fold as the heterology increased to 14%. These results suggest that discrimination of homology from heterology occurs during both pairing and strand exchange. PMID- 9342329 TI - Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription in vitro. AB - RNA polymerase I (pol I) is a nuclear enzyme whose function is to transcribe the duplicated genes encoding the precursor of the three largest ribosomal RNAs. We report a cell-free system from broccoli (Brassica oleracea) inflorescence that supports promoter-dependent RNA pol I transcription in vitro. The transcription system was purified extensively by DEAE-Sepharose, Biorex 70, Sephacryl S300, and Mono Q chromatography. Activities required for pre-rRNA transcription copurified with the polymerase on all four columns, suggesting their association as a complex. Purified fractions programmed transcription initiation from the in vivo start site and utilized the same core promoter sequences required in vivo. The complex was not dissociated in 800 mM KCl and had a molecular mass of nearly 2 MDa based on gel filtration chromatography. The most highly purified fractions contain approximately 30 polypeptides, two of which were identified immunologically as RNA polymerase subunits. These data suggest that the occurrence of a holoenzyme complex is probably not unique to the pol II system but may be a general feature of eukaryotic nuclear polymerases. PMID- 9342330 TI - Growth and viability of macrophages continuously stimulated to produce nitric oxide. AB - Deregulated production of nitric oxide (NO) has been implicated in the development of certain human diseases, including cancer. We sought to assess the damaging potential of NO produced under long-term conditions through the development of a suitable model cell culture system. In this study, we report that when murine macrophage-like RAW264.7 cells were exposed continuously to bacterial lipopolysaccharide (LPS) or mouse recombinant interferon-gamma (IFN gamma) over periods of 21-23 days, they continued to grow, but with doubling times 2 to 4 times, respectively, longer than the doubling time of unstimulated cells. Stimulated cells produced NO at rates of 30 to 70 nmol per million cells per day throughout the stimulation period. Within 24 hr after removal of stimulant, cells resumed exponential growth. Simultaneous exposure to LPS and IFN gamma resulted in decreased cell number, which persisted for 2 days after removal of the stimulants. Exponential growth was attained only after an additional 4 days. Addition of N-methyl-L-arginine (NMA), an NO synthase inhibitor, to the medium inhibited NO production by 90% of all stimulated cells, partially reduced doubling time of cells stimulated with LPS or IFN-gamma, and partially increased viability and growth rates in those exposed to both LPS and IFN-gamma. However, when incubated with LPS and IFN-gamma at low densities both in the presence and in the absence of NMA, cells grew at a rate slower than that of unstimulated cells, with no cell death, and they resumed exponential growth 24 hr after removal of stimulants. Results from cell density experiments suggest that macrophages are protected from intracellularly generated NO; much of the NO damaging activity occurred outside of the producer cells. Collectively, results presented in this study suggest that the type of cellular toxicity observed in macrophages is markedly influenced by rate of exposure to NO: at low rates of exposure, cells exhibit slower growth; at higher rates, cells begin to die; at even higher rates, cells undergo growth arrest or die. The ability of RAW264.7 cells to produce NO over many cell generations makes the cell line a useful system for the study of other aspects of cellular damage, including genotoxicity, resulting from exposure to NO under long-term conditions. PMID- 9342331 TI - Conformation of coenzyme pyrroloquinoline quinone and role of Ca2+ in the catalytic mechanism of quinoprotein methanol dehydrogenase. AB - The ab initio structures of 2,7,9-tricarboxypyrroloquinoline quinone (PQQ), semiquinone (PQQH), and dihydroquinone (PQQH2) have been determined and compared with ab initio structures of the (PQQ)Ca2+, (PQQH)Ca2+, and (PQQH2)Ca2+ complexes as well as the x-ray structure of (PQQ)Ca2+ bound at the active site of the methanol dehydrogenase (MDH) of methyltropic bacteria. Plausible mechanisms for the MDH oxidation of methanol involving the (PQQ)Ca2+ complex are explored via ab initio computations and discussed. Considering the reaction of methanol with PQQ in the absence of Ca2+, nucleophilic addition of methanol to the PQQ C-5 carbonyl followed by a retro-ene elimination is deemed unlikely due to large energy barrier. A much more favorable disposition of the methanol C-5 adduct to provide formaldehyde involves proton ionization of the intermediate followed by elimination of methoxide concerted with hydride transfer to the oxygen of the C-4 carbonyl. Much the same transition state is reached if one searches for the transition state beginning with Asp-303-CO2-general-base removal of the methanol proton of the (PQQ)Ca2+O(H)CH3 complex concerted with hydride transfer to the oxygen at C-4. For such a mechanism the role of the Ca2+ moiety would be to (i) contribute to the formation of the ES complex (ii) provide a modest decrease in the pKa of methanol substrate,; and (iii) polarize the oxygen at C-5. PMID- 9342332 TI - Molding a peptide into an RNA site by in vivo peptide evolution. AB - Short peptides corresponding to the arginine-rich domains of several RNA-binding proteins are able to bind to their specific RNA sites with high affinities and specificities. In the case of the HIV-1 Rev-Rev response element (RRE) complex, the peptide forms a single alpha-helix that binds deeply in a widened, distorted RNA major groove and makes a substantial set of base-specific and backbone contacts. Using a reporter system based on antitermination by the bacteriophage lambda N protein, it has been possible to identify novel arginine-rich peptides from combinatorial libraries that recognize the RRE with affinities and specificities similar to Rev but that appear to bind in nonhelical conformations. Here we have used codon-based mutagenesis to evolve one of these peptides, RSG-1, into an even tighter binder. After two rounds of evolution, RSG-1.2 bound the RRE with 7-fold higher affinity and 15-fold higher specificity than the wild-type Rev peptide, and in vitro competition experiments show that RSG-1.2 completely displaces the intact Rev protein from the RRE at low peptide concentrations. By fusing RRE-binding peptides to the activation domain of HIV-1 Tat, we show that the peptides can deliver Tat to the RRE site and activate transcription in mammalian cells, and more importantly, that the fusion proteins can inhibit the activity of Rev in chloramphenicol acetyltransferase reporter assays. The evolved peptides contain proline and glutamic acid mutations near the middle of their sequences and, despite the presence of a proline, show partial alpha-helix formation in the absence of RNA. These directed evolution experiments illustrate how readily complex peptide structures can be evolved within the context of an RNA framework, perhaps reflecting how early protein structures evolved in an "RNA world." PMID- 9342333 TI - Participation of the nuclear cap binding complex in pre-mRNA 3' processing. AB - Communication between the 5' and 3' ends is a common feature of several aspects of eukaryotic mRNA metabolism. In the nucleus, the pre-mRNA 5' end is bound by the nuclear cap binding complex (CBC). This RNA-protein complex plays an active role in both splicing and RNA export. We provide evidence for participation of CBC in the processing of the 3' end of the message. Depletion of CBC from HeLa cell nuclear extract strongly reduced the endonucleolytic cleavage step of the cleavage and polyadenylation process. Cleavage was restored by addition of recombinant CBC. CBC depletion was found to reduce the stability of poly(A) site cleavage complexes formed in nuclear extract. We also provide evidence that the communication between the 5' and 3' ends of the pre-mRNA during processing is mediated by the physical association of the CBC/cap complex with 3' processing factors bound at the poly(A) site. These observations, along with previous data on the function of CBC in splicing, illustrate the key role played by CBC in pre mRNA recognition and processing. The data provides further support for the hypothesis that pre-mRNAs and mRNAs may exist and be functional in the form of "closed-loops," due to interactions between factors bound at their 5' and 3' ends. PMID- 9342335 TI - Molecular switch in signal transduction: reaction paths of the conformational changes in ras p21. AB - Conformational changes in ras p21 triggered by the hydrolysis of GTP play an essential role in the signal transduction pathway. The path for the conformational change is determined by molecular dynamics simulation with a holonomic constraint directing the system from the known GTP-bound structure (with the gamma-phosphate removed) to the GDP-bound structure. The simulation is done with a shell of water molecules surrounding the protein. In the switch I region, the side chain of Tyr-32, which undergoes a large displacement, moves through the space between loop 2 and the rest of the protein, rather than on the outside of the protein. As a result, the charged residues Glu-31 and Asp-33, which interact with Raf in the homologous RafRBD-Raps complex, remain exposed during the transition. In the switch II region, the conformational changes of alpha2 and loop 4 are strongly coupled. A transient hydrogen bonding complex between Arg-68 and Tyr-71 in the switch II region and Glu-37 in switch I region stabilizes the intermediate conformation of alpha2 and facilitates the unwinding of a helical turn of alpha2 (residues 66-69), which in turn permits the larger scale motion of loop 4. Hydrogen bond exchange between the protein and solvent molecules is found to be important in the transition. Possible functional implications of the results are discussed. PMID- 9342334 TI - Both N-terminal myosin-binding and C-terminal actin-binding sites on smooth muscle caldesmon are required for caldesmon-mediated inhibition of actin filament velocity. AB - It has been suggested that the tethering caused by binding of the N-terminal region of smooth muscle caldesmon (CaD) to myosin and its C-terminal region to actin contributes to the inhibition of actin-filament movement over myosin heads in an in vitro motility assay. However, direct evidence for this assumption has been lacking. In this study, analysis of baculovirus-generated N-terminal and C terminal deletion mutants of chicken-gizzard CaD revealed that the major myosin binding site on the CaD molecule resides in a 30-amino acid stretch between residues 24 and 53, based on the very low level of binding of CaDDelta24-53 lacking the residues 24-53 to myosin compared with the level of binding of CaDDelta54-85 missing the adjacent residues 54-85 or of the full-length CaD. As expected, deletion of the region between residues 24 and 53 or between residues 54 and 85 had no effect on either actin-binding or inhibition of actomyosin ATPase activity. Deletion of residues 24-53 nearly abolished the ability of CaD to inhibit actin filament velocity in the in vitro motility experiments, whereas CaDDelta54-85 strongly inhibited actin filament velocity in a manner similar to that of full-length CaD. Moreover, CaD1-597, which lacks the major actin-binding site(s), did not inhibit actin-filament velocity despite the presence of the major myosin-binding site. These data provide direct evidence for the inhibition of actin filament velocity in the in vitro motility assay caused by the tethering of myosin to actin through binding of both the CaD N-terminal region to myosin and the C-terminal region to actin. PMID- 9342336 TI - A structural census of the current population of protein sequences. AB - We examine the occurrence of the approximately 300 known protein folds in different groups of organisms. To do this, we characterize a large fraction of the currently known protein sequences ( approximately 140,000) in structural terms, by matching them to known structures via sequence comparison (or by secondary-structure class prediction for those without structural homologues). Overall, we find that an appreciable fraction of the known folds are present in each of the major groups of organisms (e.g., bacteria and eukaryotes share 156 of 275 folds), and most of the common folds are associated with many families of nonhomologous sequences (i.e., >10 sequence families for each common fold). However, different groups of organisms have characteristically distinct distributions of folds. So, for instance, some of the most common folds in vertebrates, such as globins or zinc fingers, are rare or absent in bacteria. Many of these differences in fold usage are biologically reasonable, such as the folds of metabolic enzymes being common in bacteria and those associated with extracellular transport and communication being common in animals. They also have important implications for database-based methods for fold recognition, suggesting that an unknown sequence from a plant is more likely to have a certain fold (e.g., a TIM barrel) than an unknown sequence from an animal. PMID- 9342337 TI - Preferential exclusion of sucrose from recombinant interleukin-1 receptor antagonist: role in restricted conformational mobility and compaction of native state. AB - Understanding the mechanism for sucrose-induced protein stabilization is important in many diverse fields, ranging from biochemistry and environmental physiology to pharmaceutical science. Timasheff and Lee [Lee, J. C. & Timasheff, S. N. (1981) J. Biol. Chem. 256, 7193-7201] have established that thermodynamic stabilization of proteins by sucrose is due to preferential exclusion of the sugar from the protein's surface, which increases protein chemical potential. The current study measures the preferential exclusion of 1 M sucrose from a protein drug, recombinant interleukin 1 receptor antagonist (rhIL-1ra). It is proposed that the degree of preferential exclusion and increase in chemical potential are directly proportional to the protein surface area and that, hence, the system will favor the protein state with the smallest surface area. This mechanism explains the observed sucrose-induced restriction of rhIL-1ra conformational fluctuations, which were studied by hydrogen-deuterium exchange and cysteine reactivity measurements. Furthermore, infrared spectroscopy of rhlL-1ra suggested that a more ordered native conformation is induced by sucrose. Electron paramagnetic resonance spectroscopy demonstrated that in the presence of sucrose, spin-labeled cysteine 116 becomes more buried in the protein's interior and that the hydrodynamic diameter of the protein is reduced. The preferential exclusion of sucrose from the protein and the resulting shift in the equilibrium between protein states toward the most compact conformation account for sucrose-induced effects on rhIL-1ra. PMID- 9342339 TI - Assigning folds to the proteins encoded by the genome of Mycoplasma genitalium. AB - A crucial step in exploiting the information inherent in genome sequences is to assign to each protein sequence its three-dimensional fold and biological function. Here we describe fold assignment for the proteins encoded by the small genome of Mycoplasma genitalium. The assignment was carried out by our computer server (http://www.doe-mbi.ucla.edu/people/frsvr/ frsvr. html), which assigns folds to amino acid sequences by comparing sequence-derived predictions with known structures. Of the total of 468 protein ORFs, 103 (22%) can be assigned a known protein fold with high confidence, as cross-validated with tests on known structures. Of these sequences, 75 (16%) show enough sequence similarity to proteins of known structure that they can also be detected by traditional sequence-sequence comparison methods. That is, the difference of 28 sequences (6%) are assignable by the sequence-structure method of the server but not by current sequence-sequence methods. Of the remaining 78% of sequences in the genome, 18% belong to membrane proteins and the remaining 60% cannot be assigned either because these sequences correspond to no presently known fold or because of insensitivity of the method. At the current rate of determination of new folds by x-ray and NMR methods, extrapolation suggests that folds will be assigned to most soluble proteins in the next decade. PMID- 9342340 TI - Mechanical separation of the complementary strands of DNA. AB - We describe the mechanical separation of the two complementary strands of a single molecule of bacteriophage lambda DNA. The 3' and 5' extremities on one end of the molecule are pulled progressively apart, and this leads to the opening of the double helix. The typical forces along the opening are in the range of 10-15 pN. The separation force signal is shown to be related to the local GC vs. AT content along the molecule. Variations of this content on a typical scale of 100 500 bases are presently detected. PMID- 9342338 TI - Effects of extracellular Ca2+ concentration on hair-bundle stiffness and gating spring integrity in hair cells. AB - When a hair cell is stimulated by positive deflection of its hair bundle, increased tension in gating springs opens transduction channels, permitting cations to enter stereocilia and depolarize the cell. Ca2+ is thought to be required in mechanoelectrical transduction, for exposure of hair bundles to Ca2+ chelators eliminates responsiveness by disrupting tip links, filamentous interstereociliary connections that probably are the gating springs. Ca2+ also participates in adaptation to stimuli by controlling the activity of a molecular motor that sets gating-spring tension. Using a flexible glass fiber to measure hair-bundle stiffness, we investigated the effect of Ca2+ concentration on stiffness before and after the disruption of gating springs. The stiffness of intact hair bundles depended nonmonotonically on the extracellular Ca2+ concentration; the maximal stiffness of approximately 1200 microN.m-1 occurred when bundles were bathed in solutions containing 250 microM Ca2+, approximately the concentration found in frog endolymph. For cells exposed to solutions with sufficient chelator capacity to reduce the Ca2+ concentration below approximately 100 nM, hair-bundle stiffness fell to approximately 200 microN.m-1 and no longer exhibited Ca2+-dependent changes. Because cells so treated lost mechanoelectrical transduction, we attribute the reduction in bundle stiffness to tip-link disruption. The results indicate that gating springs are not linearly elastic; instead, they stiffen with increased strain, which rises with adaptation-motor activity at the physiological extracellular Ca2+ concentration. PMID- 9342341 TI - Human deoxycytidine kinase is located in the cell nucleus. AB - Human deoxyribonucleoside kinases are required for the pharmacological activity of several clinically important anticancer and antiviral nucleoside analogs. Human deoxycytidine kinase and thymidine kinase 1 are described as cytosolic enzymes in the literature, whereas human deoxyguanosine kinase and thymidine kinase 2 are believed to be located in the mitochondria. We expressed the four human deoxyribonucleoside kinases as fusion proteins with the green fluorescent protein to study their intracellular locations in vivo. Our data showed that the human deoxycytidine kinase is located in the cell nucleus and the human deoxyguanosine kinase is located in the mitochondria. The fusion proteins between green fluorescent protein and thymidine kinases 1 and 2 were both predominantly located in the cytosol. Site-directed mutagenesis of a putative nuclear targeting signal, identified in the primary structure of deoxycytidine kinase, completely abolished nuclear import of the protein. Reconstitution of a deoxycytidine kinase deficient cell line with the wild-type nuclear or the mutant cytosolic enzymes both restored sensitivity toward anticancer nucleoside analogs. This paper reports that a deoxyribonucleoside kinase is located in the cell nucleus and we discuss the implications for deoxyribonucleotide synthesis and phosphorylation of nucleoside analogs. PMID- 9342342 TI - Control of membrane phosphatidylcholine biosynthesis by diacylglycerol levels in neuronal cells undergoing neurite outgrowth. AB - Phospholipids are the major components of cell membranes and are required for cellular growth. We studied membrane phosphatidylcholine (PtdCho) biosynthesis in neuronal cells undergoing neurite outgrowth, by using PC12 cells as a model system. When neurite outgrowth was induced by exposing PC12 cells to nerve growth factor for 2 and 4 days, the amounts of [14C]choline incorporated into [14C]phosphatidylcholine per cell (i.e., per DNA) increased approximately 5- and 10-fold, respectively, as compared with control cells, reflecting increases in the rate of PtdCho biosynthesis. [14C]choline uptake was not affected. Analysis of the three major PtdCho biosynthetic enzymes showed that the activity of CDPcholine:1,2-diacylglycerol cholinephosphotransferase was increased by approximately 50% after nerve growth factor treatment, but the activities of choline kinase or choline-phosphate cytidylyltransferase were unaltered; the cholinephosphotransferase displayed a high Km value ( approximately 1,200 microM) for diacylglycerol. Moreover, free cellular diacylglycerol levels increased by approximately 1.5- and 4-fold on the second and fourth days, respectively. These data indicate that PtdCho biosynthesis is enhanced when PC12 cells sprout neurites, and the enhancement is mediated primarily by changes in cholinephosphotransferase activity and its saturation with diacylglycerol. This suggests a novel regulatory role for diacylglycerol in membrane phospholipid biosynthesis. PMID- 9342343 TI - Activation of distinct caspase-like proteases by Fas and reaper in Drosophila cells. AB - The cytoplasmic region of Fas, a mammalian death factor receptor, shares a limited homology with reaper, an apoptosis-inducing protein in Drosophila. Expression of either the Fas cytoplasmic region (FasC) or of reaper in Drosophila cells caused cell death. The death process induced by FasC or reaper was inhibited by crmA or p35, suggesting that its death process is mediated by caspase-like proteases. Both Ac-YVAD aldehyde and Ac-DEVD aldehyde, specific inhibitors of caspase 1- and caspase 3-like proteases, respectively, inhibited the FasC-induced death of Drosophila cells. However, the cell death induced by reaper was inhibited by Ac-DEVD aldehyde, but not by Ac-YVAD aldehyde. A caspase 1-like protease activity that preferentially recognizes the YVAD sequence gradually increased in the cytosolic fraction of the FasC-activated cells, whereas the caspase 3-like protease activity recognizing the DEVD sequence was observed in the reaper-activated cells. Partial purification and biochemical characterization of the proteases indicated that there are at least three distinct caspase-like proteases in Drosophila cells, which are differentially activated by FasC and reaper. The conservation of the Fas-death signaling pathway in Drosophila cells, which is distinct from that for reaper, may indicate that cell death in Drosophila is controlled not only by the reaper suicide gene, but also by a Fas-like killer gene. PMID- 9342344 TI - Proteinase inhibitors I and II from potatoes specifically block UV-induced activator protein-1 activation through a pathway that is independent of extracellular signal-regulated kinases, c-Jun N-terminal kinases, and P38 kinase. AB - Solar UV irradiation is the causal factor for the increasing incidence of human skin carcinomas. The activation of the transcription factor activator protein-1 (AP-1) has been shown to be responsible for the tumor promoter action of UV light in mammalian cells. We demonstrate that proteinase inhibitor I (Inh I) and II (Inh II) from potato tubers, when applied to mouse epidermal JB6 cells, block UV induced AP-1 activation. The inhibition appears to be specific for UV-induced signal transduction for AP-1 activation, because these inhibitors did not block UV-induced p53 activation nor did they exhibit any significant influence on epidermal growth factor-induced AP-1 transactivation. Furthermore, the inhibition of UV-induced AP-1 activity occurs through a pathway that is independent of extracellular signal-regulated kinases and c-Jun N-terminal kinases as well as P38 kinases. Considering the important role of AP-1 in tumor promotion, it is possible that blocking UV-induced AP-1 activity by Inh I or Inh II may be functionally linked to irradiation-induced cell transformation. PMID- 9342345 TI - Conversion of cysteine to formylglycine: a protein modification in the endoplasmic reticulum. AB - In sulfatases a Calpha-formylglycine residue is found at a position where their cDNA sequences predict a cysteine residue. In multiple sulfatase deficiency, an inherited lysosomal storage disorder, catalytically inactive sulfatases are synthesized which retain the cysteine residue, indicating that the Calpha formylglycine residue is required for sulfatase activity. Using in vitro translation in the absence or presence of transport competent microsomes we found that newly synthesized sulfatase polypeptides carry a cysteine residue and that the oxidation of its thiol group to an aldehyde is catalyzed in the endoplasmic reticulum. A linear sequence of 16 residues surrounding the Cys-69 in arylsulfatase A is sufficient to direct the oxidation. This novel protein modification occurs after or at a late stage of cotranslational protein translocation into the endoplasmic reticulum when the polypeptide is not yet folded to its native structure. PMID- 9342346 TI - Abnormalities of pancreatic islets by targeted expression of a dominant-negative KATP channel. AB - ATP-sensitive K+ (KATP) channels are known to play important roles in various cellular functions, but the direct consequences of disruption of KATP channel function are largely unknown. We have generated transgenic mice expressing a dominant-negative form of the KATP channel subunit Kir6.2 (Kir6.2G132S, substitution of glycine with serine at position 132) in pancreatic beta cells. Kir6.2G132S transgenic mice develop hypoglycemia with hyperinsulinemia in neonates and hyperglycemia with hypoinsulinemia and decreased beta cell population in adults. KATP channel function is found to be impaired in the beta cells of transgenic mice with hyperglycemia. In addition, both resting membrane potential and basal calcium concentrations are shown to be significantly elevated in the beta cells of transgenic mice. We also found a high frequency of apoptotic beta cells before the appearance of hyperglycemia in the transgenic mice, suggesting that the KATP channel might play a significant role in beta cell survival in addition to its role in the regulation of insulin secretion. PMID- 9342347 TI - Mouse Eya genes are expressed during limb tendon development and encode a transcriptional activation function. AB - Vertebrate limb tendons are derived from connective cells of the lateral plate mesoderm. Some of the developmental steps leading to the formation of vertebrate limb tendons have been previously identified; however, the molecular mechanisms responsible for tendinous patterning and maintenance during embryogenesis are largely unknown. The eyes absent (eya) gene of Drosophila encodes a novel nuclear protein of unknown molecular function. Here we show that Eya1 and Eya2, two mouse homologues of Drosophila eya, are expressed initially during limb development in connective tissue precursor cells. Later in limb development, Eya1 and Eya2 expression is associated with cell condensations that form different sets of limb tendons. Eya1 expression is largely restricted to flexor tendons, while Eya2 is expressed in the extensor tendons and ligaments of the phalangeal elements of the limb. These data suggest that Eya genes participate in the patterning of the distal tendons of the limb. To investigate the molecular functions of the Eya gene products, we have analyzed whether the highly divergent PST (proline-serine threonine)-rich N-terminal regions of Eya1-3 function as transactivation domains. Our results demonstrate that Eya gene products can act as transcriptional activators, and they support a role for this molecular function in connective tissue patterning. PMID- 9342348 TI - Xenopus Zic3, a primary regulator both in neural and neural crest development. AB - Xenopus Zic3 is a Xenopus homologue of mouse Zic and Drosophila pair-rule gene, odd-paired. We show here that Zic3 has significant roles both in neural and neural crest development in Xenopus embryo. Expression of Zic3 is first detected in prospective neural plate region at gastrulation. Onset of the expression was earlier than most proneural genes and followed chordin expression. The expression was induced by blockade of BMP4 signal. Overexpression of Zic3 resulted in hyperplastic neural and neural crest derived tissue. In animal cap explant, the overexpression of Zic3 induced expression of all the proneural genes and neural crest marker genes. These findings suggest that Zic3 can determine the ectodermal cell fate and promote the earliest step of neural and neural crest development. PMID- 9342349 TI - Identification of active sites in amidase: evolutionary relationship between amide bond- and peptide bond-cleaving enzymes. AB - Mainly based on various inhibitor studies previously performed, amidases came to be regarded as sulfhydryl enzymes. Not completely satisfied with this generally accepted interpretation, we performed a series of site-directed mutagenesis studies on one particular amidase of Rhodococcus rhodochrous J1 that was involved in its nitrile metabolism. For these experiments, the recombinant amidase was produced as the inclusion body in Escherichia coli to greatly facilitate its recovery and subsequent purification. With regard to the presumptive active site residue Cys203, a Cys203 --> Ala mutant enzyme still retained 11.5% of the original specific activity. In sharp contrast, substitutions in certain other positions in the neighborhood of Cys203 had a far more dramatic effect on the amidase. Glutamic acid substitution of Asp191 reduced the specific activity of the mutant enzyme to 1.33% of the wild-type activity. Furthermore, Asp191 --> Asn substitution as well as Ser195 --> Ala substitution completely abolished the specific activity. It would thus appear that, among various conserved residues residing within the so-called signature sequence common to all amidases, the real active site residues are Asp191 and Ser195 rather than Cys203. Inasmuch as an amide bond (CO-NH2) in the amide substrate is not too far structurally removed from a peptide bond (CO-NH-), the signature sequences of various amidases were compared with the active site sequences of various types of proteases. It was found that aspartic acid and serine residues corresponding to Asp191 and Ser195 of the Rhodococcus amidase are present within the active site sequences of aspartic proteinases, thus suggesting the evolutionary relationship between the two. PMID- 9342350 TI - Convergent evolution of apolipoprotein(a) in primates and hedgehog. AB - Apolipoprotein(a) [apo(a)] is the distinguishing protein component of lipoprotein(a), a major inherited risk factor for atherosclerosis. Human apo(a) is homologous to plasminogen. It contains from 15 to 50 repeated domains closely related to plasminogen kringle four, plus single kringle five-like and inactive protease-like domains. This expressed gene is confined to a subset of primates. Although most mammals lack apo(a), hedgehogs produce an apo(a)-like protein composed of highly repeated copies of a plasminogen kringle three-like domain, with complete absence of protease domain sequences. Both human and hedgehog apo(a)-like proteins form covalently linked lipoprotein particles that can bind to fibrin and other substrates shared with plasminogen. DNA sequence comparisons and phylogenetic analysis indicate that the human type of apo(a) evolved from a duplicated plasminogen gene during recent primate evolution. In contrast, the kringle three-based type of apo(a) evolved from an independent duplication of the plasminogen gene approximately 80 million years ago. In a type of convergent evolution, the plasminogen gene has been independently remodeled twice during mammalian evolution to produce similar forms of apo(a) in two widely divergent groups of species. PMID- 9342352 TI - Evolution of gilled mushrooms and puffballs inferred from ribosomal DNA sequences. AB - Homobasidiomycete fungi display many complex fruiting body morphologies, including mushrooms and puffballs, but their anatomical simplicity has confounded efforts to understand the evolution of these forms. We performed a comprehensive phylogenetic analysis of homobasidiomycetes, using sequences from nuclear and mitochondrial ribosomal DNA, with an emphasis on understanding evolutionary relationships of gilled mushrooms and puffballs. Parsimony-based optimization of character states on our phylogenetic trees suggested that strikingly similar gilled mushrooms evolved at least six times, from morphologically diverse precursors. Approximately 87% of gilled mushrooms are in a single lineage, which we call the "euagarics." Recently discovered 90 million-year-old fossil mushrooms are probably euagarics, suggesting that (i) the origin of this clade must have occurred no later than the mid-Cretaceous and (ii) the gilled mushroom morphology has been maintained in certain lineages for tens of millions of years. Puffballs and other forms with enclosed spore-bearing structures (Gasteromycetes) evolved at least four times. Derivation of Gasteromycetes from forms with exposed spore bearing structures (Hymenomycetes) is correlated with repeated loss of forcible spore discharge (ballistospory). Diverse fruiting body forms and spore dispersal mechanisms have evolved among Gasteromycetes. Nevertheless, it appears that Hymenomycetes have never been secondarily derived from Gasteromycetes, which suggests that the loss of ballistospory has constrained evolution in these lineages. PMID- 9342353 TI - Origin and evolution of the slime molds (Mycetozoa) AB - The Mycetozoa include the cellular (dictyostelid), acellular (myxogastrid), and protostelid slime molds. However, available molecular data are in disagreement on both the monophyly and phylogenetic position of the group. Ribosomal RNA trees show the myxogastrid and dictyostelid slime molds as unrelated early branching lineages, but actin and beta-tubulin trees place them together as a single coherent (monophyletic) group, closely related to the animal-fungal clade. We have sequenced the elongation factor-1alpha genes from one member of each division of the Mycetozoa, including Dictyostelium discoideum, for which cDNA sequences were previously available. Phylogenetic analyses of these sequences strongly support a monophyletic Mycetozoa, with the myxogastrid and dictyostelid slime molds most closely related to each other. All phylogenetic methods used also place this coherent Mycetozoan assemblage as emerging among the multicellular eukaryotes, tentatively supported as more closely related to animals + fungi than are green plants. With our data there are now three proteins that consistently support a monophyletic Mycetozoa and at least four that place these taxa within the "crown" of the eukaryote tree. We suggest that ribosomal RNA data should be more closely examined with regard to these questions, and we emphasize the importance of developing multiple sequence data sets. PMID- 9342355 TI - Highly conservative reciprocal translocations formed by apparent joining of exchanged DNA double-strand break ends. AB - Chromosomal translocations induced by ionizing radiation and radiomimetic drugs are thought to arise by incorrect joining of DNA double-strand breaks. To dissect such misrepair events at a molecular level, large-scale, bleomycin-induced rearrangements in the aprt gene of Chinese hamster ovary D422 cells were mapped, the breakpoints were sequenced, and the original non-aprt parental sequences involved in each rearrangement were recovered from nonmutant cells. Of seven rearrangements characterized, six were reciprocal exchanges between aprt and unrelated sequences. Consistent with a mechanism involving joining of exchanged double-strand break ends, there was, in most cases, no homology between the two parental sequences, no overlap in sequences retained at the two newly formed junctions, and little or no loss of parental sequences (usually /= 257 micromol/liter, vs. 22 (9.2%) normals (P = 0.0005). Of those with the normal homozygous UDPGT1 genotype, the incidence of hyperbilirubinemia was similar in G 6-PD-deficients and controls (9.7% and 9.9%). In contrast, in the G-6-PD deficient neonates, those with the heterozygous or homozygous variant UDPGT1 genotype had a higher incidence of hyperbilirubinemia than corresponding controls (heterozygotes: 31.6% vs. 6.7%, P < 0.0001; variant homozygotes: 50% vs. 14.7%, P = 0.02). Among G-6-PD-deficient infants the incidence of hyperbilirubinemia was greater in those with the heterozygous (31.6%, P = 0.006) or variant homozygous (50%, P = 0.003) UDPGT1 genotype than in normal homozygotes. In contrast, among those normal for G-6-PD, the UDPGT1 polymorphism had no significant effect (heterozygotes: 6.7%; variant homozygotes: 14.7%). Thus, neither G-6-PD deficiency nor the variant UDPGT1 promoter, alone, increased the incidence of hyperbilirubinemia, but both in combination did. This gene interaction may serve as a paradigm of the interaction of benign genetic polymorphisms in the causation of disease. PMID- 9342375 TI - Prolonged production of NADPH oxidase-corrected granulocytes after gene therapy of chronic granulomatous disease. AB - Little is known about the potential for engraftment of autologous hematopoietic stem cells in human adults not subjected to myeloablative conditioning regimens. Five adult patients with the p47(phox) deficiency form of chronic granulomatous disease received intravenous infusions of autologous CD34(+) peripheral blood stem cells (PBSCs) that had been transduced ex vivo with a recombinant retrovirus encoding normal p47(phox). Although marrow conditioning was not given, functionally corrected granulocytes were detectable in peripheral blood of all five patients. Peak correction occurred 3-6 weeks after infusion and ranged from 0.004 to 0.05% of total peripheral blood granulocytes. Corrected cells were detectable for as long as 6 months after infusion in some individuals. Thus, prolonged engraftment of autologous PBSCs and continued expression of the transduced gene can occur in adults without conditioning. This trial also piloted the use of animal protein-free medium and a blood-bank-compatible closed system of gas-permeable plastic containers for culture and transduction of the PBSCs. These features enhance the safety of PBSCs directed gene therapy. PMID- 9342376 TI - Cloning and characterization of the Flavobacterium johnsoniae (Cytophaga johnsonae) gliding motility gene, gldA. AB - The mechanism of bacterial gliding motility (active movement over surfaces without the aid of flagella) is not known. A large number of nonmotile mutants of the gliding bacterium Flavobacterium johnsoniae (Cytophaga johnsonae) have been previously isolated, and genetic techniques to analyze these mutants have recently been developed. We complemented a nonmotile mutant of F. johnsoniae (UW102-09) with a library of wild-type DNA by using the shuttle cosmid pCP17. The complementing plasmid (pCP100) contained an insert of 13 kbp, and restored motility to 4 of 61 independently isolated nonmotile mutants. A 1.3-kbp fragment that encompassed a single ORF, gldA, complemented all four mutants. Disruption of the chromosomal copy of gldA in wild-type F. johnsoniae UW101 eliminated gliding motility. The predicted protein produced by gldA has strong sequence similarity to ATP binding cassette transport proteins. PMID- 9342378 TI - Operative GABAergic inhibition in hippocampal CA1 pyramidal neurons in experimental epilepsy. AB - Patch-clamp recordings of CA1 interneurons and pyramidal cells were performed in hippocampal slices from kainate- or pilocarpine-treated rat models of temporal lobe epilepsy. We report that gamma-aminobutyric acid (GABA)ergic inhibition in pyramidal neurons is still functional in temporal lobe epilepsy because: (i) the frequency of spontaneous GABAergic currents is similar to that of control and (ii) focal electrical stimulation of interneurons evokes a hyperpolarization that prevents the generation of action potentials. In paired recordings of interneurons and pyramidal cells, synchronous interictal activities were recorded. Furthermore, large network-driven GABAergic inhibitory postsynaptic currents were present in pyramidal cells during interictal discharges. The duration of these interictal discharges was increased by the GABA type A antagonist bicuculline. We conclude that GABAergic inhibition is still present and functional in these experimental models and that the principal defect of inhibition does not lie in a complete disconnection of GABAergic interneurons from their glutamatergic inputs. PMID- 9342379 TI - Loss of haloperidol induced gene expression and catalepsy in protein kinase A deficient mice. AB - The antipsychotic drug, haloperidol, elicits the expression of neurotensin and c fos mRNA in the dorsal lateral region of the striatum and produces an acute cataleptic response in rodents that correlates with the motor side effects of haloperidol in humans. Mice harboring a targeted disruption of the RIIbeta subunit of protein kinase A have a profound deficit in cAMP-stimulated kinase activity in the striatum. When treated with haloperidol, RIIbeta mutant mice fail to induce either c-fos or neurotensin mRNA and the acute cataleptic response is blocked. However, both wild-type and mutant mice become cataleptic when neurotensin peptide is directly injected into the lateral ventricle, demonstrating that the kinase deficiency does not interfere with the action of neurotensin but rather its synthesis and release. These results establish a direct role for protein kinase A as a mediator of haloperidol induced gene induction and cataleptic behavior. PMID- 9342377 TI - Probing the assembly of transcription initiation complexes through changes in sigmaN protease sensitivity. AB - The alternative bacterial sigmaN RNA polymerase holoenzyme binds promoters as a transcriptionally inactive complex that is activated by enhancer-binding proteins. Little is known about how sigma factors respond to their ligands or how the responses lead to transcription. To examine the liganded state of sigmaN, the assembly of end-labeled Klebsiella pneumoniae sigmaN into holoenzyme, closed promoter complexes, and initiated transcription complexes was analyzed by enzymatic protein footprinting. V8 protease-sensitive sites in free sigmaN were identified in the acidic region II and bordering or within the minimal DNA binding domain. Interaction with core RNA polymerase prevented cleavage at noncontiguous sites in region II and at some DNA binding domain sites, probably resulting from conformational changes. Formation of closed complexes resulted in further protections within the DNA binding domain, suggesting close contact to promoter DNA. Interestingly, residue E36 becomes sensitive to proteolysis in initiated transcription complexes, indicating a conformational change in holoenzyme during initiation. Residue E36 is located adjacent to an element involved in nucleating strand separation and in inhibiting polymerase activity in the absence of activation. The sensitivity of E36 may reflect one or both of these functions. Changing patterns of protease sensitivity strongly indicate that sigmaN can adjust conformation upon interaction with ligands, a property likely important in the dynamics of the protein during transcription initiation. PMID- 9342380 TI - The Caenorhabditis elegans seven-transmembrane protein ODR-10 functions as an odorant receptor in mammalian cells. AB - The nematode Caenorhabditis elegans exhibits behavioral responses to many volatile odorants. Chemotaxis toward one such odorant, diacetyl (butanedione), requires the function of a seven-transmembrane receptor protein encoded by the odr-10 gene. To determine directly whether ODR-10 protein is an odorant receptor, it is necessary to express the protein in a heterologous system and show that it responds to diacetyl by activation of a G protein signaling pathway. Here we demonstrate that human cells expressing ODR-10 on their surfaces exhibit a transient elevation in intracellular Ca2+ levels after diacetyl application. Volatile compounds that differ from diacetyl only by the addition of a methyl group (2,3-pentanedione) or the absence of a keto group (butanone) are not ODR-10 agonists. Behavioral responses to these compounds are not dependent on odr-10 function, so ODR-10 specificity in human cells resembles in vivo specificity. The apparent affinity of ODR-10 for diacetyl observed in human cells is consistent with the diacetyl concentration ranges that allow efficient nematode chemotaxis. ODR-10 expressed in human cells also responds to two anionic compounds, pyruvate and citrate, which are metabolic precursors used for diacetyl production by certain bacterial species. Ca2+ elevation in response to ODR-10 activation is due to release from intracellular stores. PMID- 9342381 TI - Synapsin I interacts with c-Src and stimulates its tyrosine kinase activity. AB - Synapsin I is a synaptic vesicle-associated phosphoprotein that has been implicated in the formation of presynaptic specializations and in the regulation of neurotransmitter release. The nonreceptor tyrosine kinase c-Src is enriched on synaptic vesicles, where it accounts for most of the vesicle-associated tyrosine kinase activity. Using overlay, affinity chromatography, and coprecipitation assays, we have now shown that synapsin I is the major binding protein for the Src homology 3 (SH3) domain of c-Src in highly purified synaptic vesicle preparations. The interaction was mediated by the proline-rich domain D of synapsin I and was not significantly affected by stoichiometric phosphorylation of synapsin I at any of the known regulatory sites. The interaction of purified c Src and synapsin I resulted in a severalfold stimulation of tyrosine kinase activity and was antagonized by the purified c-Src-SH3 domain. Depletion of synapsin I from purified synaptic vesicles resulted in a decrease of endogenous tyrosine kinase activity. Portions of the total cellular pools of synapsin I and Src were coprecipitated from detergent extracts of rat brain synaptosomal fractions using antibodies to either protein species. The interaction between synapsin I and c-Src, as well as the synapsin I-induced stimulation of tyrosine kinase activity, may be physiologically important in signal transduction and in the modulation of the function of axon terminals, both during synaptogenesis and at mature synapses. PMID- 9342382 TI - Response-reinforcement learning is dependent on N-methyl-D-aspartate receptor activation in the nucleus accumbens core. AB - The nucleus accumbens, a site within the ventral striatum, is best known for its prominent role in mediating the reinforcing effects of drugs of abuse such as cocaine, alcohol, and nicotine. Indeed, it is generally believed that this structure subserves motivated behaviors, such as feeding, drinking, sexual behavior, and exploratory locomotion, which are elicited by natural rewards or incentive stimuli. A basic rule of positive reinforcement is that motor responses will increase in magnitude and vigor if followed by a rewarding event. It is likely, therefore, that the nucleus accumbens may serve as a substrate for reinforcement learning. However, there is surprisingly little information concerning the neural mechanisms by which appetitive responses are learned. In the present study, we report that treatment of the nucleus accumbens core with the selective competitive N-methyl-D-aspartate (NMDA) antagonist 2-amino-5 phosphonopentanoic acid (AP-5; 5 nmol/0.5 microl bilaterally) impairs response reinforcement learning in the acquisition of a simple lever-press task to obtain food. Once the rats learned the task, AP-5 had no effect, demonstrating the requirement of NMDA receptor-dependent plasticity in the early stages of learning. Infusion of AP-5 into the accumbens shell produced a much smaller impairment of learning. Additional experiments showed that AP-5 core-treated rats had normal feeding and locomotor responses and were capable of acquiring stimulus reward associations. We hypothesize that stimulation of NMDA receptors within the accumbens core is a key process through which motor responses become established in response to reinforcing stimuli. Further, this mechanism, may also play a critical role in the motivational and addictive properties of drugs of abuse. PMID- 9342383 TI - The interaction between cytoplasmic dynein and dynactin is required for fast axonal transport. AB - Fast axonal transport is characterized by the bidirectional, microtubule-based movement of membranous organelles. Cytoplasmic dynein is necessary but not sufficient for retrograde transport directed from the synapse to the cell body. Dynactin is a heteromultimeric protein complex, enriched in neurons, that binds to both microtubules and cytoplasmic dynein. To determine whether dynactin is required for retrograde axonal transport, we examined the effects of anti dynactin antibodies on organelle transport in extruded axoplasm. Treatment of axoplasm with antibodies to the p150(Glued) subunit of dynactin resulted in a significant decrease in the velocity of microtubule-based organelle transport, with many organelles bound along microtubules. We examined the molecular mechanism of the observed inhibition of motility, and we demonstrated that antibodies to p150(Glued) disrupted the binding of cytoplasmic dynein to dynactin and also inhibited the association of cytoplasmic dynein with organelles. In contrast, the anti-p150(Glued) antibodies had no effect on the binding of dynactin to microtubules nor on cytoplasmic dynein-driven microtubule gliding. These results indicate that the interaction between cytoplasmic dynein and the dynactin complex is required for the axonal transport of membrane-bound vesicles and support the hypothesis that dynactin may function as a link between the organelle, the microtubule, and cytoplasmic dynein during vesicle transport. PMID- 9342384 TI - Disruption of syntaxin-mediated protein interactions blocks neurotransmitter secretion. AB - The membrane protein syntaxin participates in several protein-protein interactions that have been implicated in neurotransmitter release. To probe the physiological importance of these interactions, we microinjected into the squid giant presynaptic terminal botulinum toxin C1, which cleaves syntaxin, and the H3 domain of syntaxin, which mediates binding to other proteins. Both reagents inhibited synaptic transmission yet did not affect the number or distribution of synaptic vesicles at the presynaptic active zone. Recombinant H3 domain inhibited the interactions between syntaxin and SNAP-25 that underlie the formation of stable SNARE complexes in vitro. These data support the notion that syntaxin mediated SNARE complexes are necessary for docked synaptic vesicles to fuse. PMID- 9342385 TI - Protein kinase C as a signal for exocytosis. AB - We have studied signaling mechanisms that stimulate exocytosis and luteinizing hormone secretion in isolated male rat pituitary gonadotropes. As judged by reverse hemolytic plaque assays, phorbol-12-myristate-13-acetate (PMA) stimulates as many gonadotropes to secrete as does gonadotropin-releasing hormone (GnRH). However, PMA and GnRH use different signaling pathways. The secretagogue action of GnRH is not very sensitive to bisindolylmaleimide I, an inhibitor of protein kinase C, but is blocked by loading cells with a calcium chelator, 1,2-bis-(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. The secretagogue action of PMA is blocked by bisindolylmaleimide I and is not very sensitive to the intracellular calcium chelator. GnRH induces intracellular calcium elevations, whereas PMA does not. As judged by amperometric measurements of quantal catecholamine secretion from dopamine- or serotonin-loaded gonadotropes, the secretagogue action of PMA develops more slowly (in several minutes) than that of GnRH. We conclude that exocytosis of secretory vesicles can be stimulated independently either by calcium elevations or by activation of protein kinase C. PMID- 9342386 TI - Recurrent excitatory postsynaptic potentials induced by synchronized fast cortical oscillations. AB - Gamma frequency (about 20-70 Hz) oscillations occur during novel sensory stimulation, with tight synchrony over distances of at least 7 mm. Synchronization in the visual system has been proposed to reflect coactivation of different parts of the visual field by a single spatially extended object. We have shown that intracortical mechanisms, including spike doublet firing by interneurons, can account for tight long-range synchrony. Here we show that synchronous gamma oscillations in two sites also can cause long-lasting (>1 hr) potentiation of recurrent excitatory synapses. Synchronous oscillations lasting >400 ms in hippocampal area CA1 are associated with an increase in both excitatory postsynaptic potential (EPSP) amplitude and action potential afterhyperpolarization size. The resulting EPSPs stabilize and synchronize a prolonged beta frequency (about 10-25 Hz) oscillation. The changes in EPSP size are not expressed during non-oscillatory behavior but reappear during subsequent gamma-oscillatory events. We propose that oscillation-induced EPSPs serve as a substrate for memory, whose expression either enhances or blocks synchronization of spatially separated sites. This phenomenon thus provides a dynamical mechanism for storage and retrieval of stimulus-specific neuronal assemblies. PMID- 9342387 TI - HOP-1, a Caenorhabditis elegans presenilin, appears to be functionally redundant with SEL-12 presenilin and to facilitate LIN-12 and GLP-1 signaling. AB - Mutant presenilins have been found to cause Alzheimer disease. Here, we describe the identification and characterization of HOP-1, a Caenorhabditis elegans presenilin that displays much more lower sequence identity with human presenilins than does the other C. elegans presenilin, SEL-12. Despite considerable divergence, HOP-1 appears to be a bona fide presenilin, because HOP-1 can rescue the egg-laying defect caused by mutations in sel-12 when hop-1 is expressed under the control of sel-12 regulatory sequences. HOP-1 also has the essential topological characteristics of the other presenilins. Reducing hop-1 activity in a sel-12 mutant background causes synthetic lethality and terminal phenotypes associated with reducing the function of the C. elegans lin-12 and glp-1 genes. These observations suggest that hop-1 is functionally redundant with sel-12 and underscore the intimate connection between presenilin activity and LIN-12/Notch activity inferred from genetic studies in C. elegans and mammals. PMID- 9342389 TI - Design of compounds that increase the absorption of polar molecules. AB - Hydrophilic drugs are often poorly absorbed when administered orally. There has been considerable interest in the possibility of using absorption enhancers to promote absorption of polar molecules across membrane surfaces. The bile acids are one of the most widely investigated classes of absorption enhancers, but there is disagreement about what features of bile acid enhancers are responsible for their efficacy. We have designed a class of glycosylated bile acid derivatives to evaluate how increasing the hydrophilicity of the steroid nucleus affects the ability to transport polar molecules across membranes. Some of the glycosylated molecules are significantly more effective than taurocholate in promoting the intestinal absorption of a range of drugs, showing that hydrophobicity is not a critical parameter in transport efficacy, as previously suggested. Furthermore, the most effective glycosylated compound is also far less damaging to membranes than the best bile acid absorption promoters, presumably because it is more hydrophilic. The results reported here show that it is possible to decouple absorption-promoting activity from membrane damage, a finding that should spark interest in the design of new compounds to facilitate the delivery of polar drugs. PMID- 9342388 TI - Defective gamma-aminobutyric acid type B receptor-activated inwardly rectifying K+ currents in cerebellar granule cells isolated from weaver and Girk2 null mutant mice. AB - Stimulation of inhibitory neurotransmitter receptors, such as gamma-aminobutyric acid type B (GABAB) receptors, activates G protein-gated inwardly rectifying K+ channels (GIRK) which, in turn, influence membrane excitability. Seizure activity has been reported in a Girk2 null mutant mouse lacking GIRK2 channels but showing normal cerebellar development as well as in the weaver mouse, which has mutated GIRK2 channels and shows abnormal development. To understand how the function of GIRK2 channels differs in these two mutant mice, we compared the G protein activated inwardly rectifying K+ currents in cerebellar granule cells isolated from Girk2 null mutant and weaver mutant mice with those from wild-type mice. Activation of GABAB receptors in wild-type granule cells induced an inwardly rectifying K+ current, which was sensitive to pertussis toxin and inhibited by external Ba2+ ions. The amplitude of the GABAB receptor-activated current was severely attenuated in granule cells isolated from both weaver and Girk2 null mutant mice. By contrast, the G protein-gated inwardly rectifying current and possibly the agonist-independent basal current appeared to be less selective for K+ ions in weaver but not Girk2 null mutant granule cells. Our results support the hypothesis that a nonselective current leads to the weaver phenotype. The loss of GABAB receptor-activated GIRK current appears coincident with the absence of GIRK2 channel protein and the reduction of GIRK1 channel protein in the Girk2 null mutant mouse, suggesting that GABAB receptors couple to heteromultimers composed of GIRK1 and GIRK2 channel subunits. PMID- 9342390 TI - Muscarinic stimulation of synaptic activity by protein kinase C is inhibited by adenosine in cultured hippocampal neurons. AB - We have studied the effect of the cholinergic agonist carbachol on the spontaneous release of glutamate in cultured rat hippocampal cells. Spontaneous excitatory postsynaptic currents (sEPSCs) through glutamatergic alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type channels were recorded by means of the patch-clamp technique. Carbachol increased the frequency of sEPSCs in a concentration-dependent manner. The kinetic properties of the sEPSCs and the amplitude distribution histograms were not affected by carbachol, arguing for a presynaptic site of action. This was confirmed by measuring the turnover of the synaptic vesicular pool by means of the fluorescent dye FM 1-43. The carbachol induced increase in sEPSC frequency was not mimicked by nicotine, but could be blocked by atropine or by pirenzepine, a muscarinic cholinergic receptor subtype M1 antagonist. Intracellular Ca2+ signals recorded with the fluorescent probe Fluo-3 indicated that carbachol transiently increased intracellular Ca2+ concentration. Since, however, carbachol still enhanced the sEPSC frequency in bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate-loaded cells, this effect could not be attributed to the rise in intracellular Ca2+ concentration. On the other hand, the protein kinase inhibitor staurosporine as well as a down-regulation of protein kinase C by prolonged treatment of the cells with 4beta-phorbol 12 myristate 13-acetate inhibited the carbachol effect. This argues for an involvement of protein kinase C in presynaptic regulation of spontaneous glutamate release. Adenosine, which inhibits synaptic transmission, suppressed the carbachol-induced stimulation of sEPSCs by a G protein-dependent mechanism activated by presynaptic A1-receptors. PMID- 9342391 TI - Two hemoglobin genes in Arabidopsis thaliana: the evolutionary origins of leghemoglobins. AB - We cloned two hemoglobin genes from Arabidopsis thaliana. One gene, AHB1, is related in sequence to the family of nonsymbiotic hemoglobin genes previously identified in a number of plant species (class 1). The second hemoglobin gene, AHB2, represents a class of nonsymbiotic hemoglobin (class 2) related in sequence to the symbiotic hemoglobin genes of legumes and Casuarina. The properties of these two hemoglobins suggest that the two families of nonsymbiotic hemoglobins may differ in function from each other and from the symbiotic hemoglobins. AHB1 is induced, in both roots and rosette leaves, by low oxygen levels. Recombinant AHB1 has an oxygen affinity so high as to make it unlikely to function as an oxygen transporter. AHB2 is expressed at a low level in rosette leaves and is low temperature-inducible. AHB2 protein has a lower affinity for oxygen than AHB1 but is similar to AHB1 in having an unusually low, pH-sensitive oxygen off-rate. PMID- 9342392 TI - Genetic control of abscisic acid biosynthesis in maize. AB - Abscisic acid (ABA), an apocarotenoid synthesized from cleavage of carotenoids, regulates seed maturation and stress responses in plants. The viviparous seed mutants of maize identify genes involved in synthesis and perception of ABA. Two alleles of a new mutant, viviparous14 (vp14), were identified by transposon mutagenesis. Mutant embryos had normal sensitivity to ABA, and detached leaves of mutant seedlings showed markedly higher rates of water loss than those of wild type. The ABA content of developing mutant embryos was 70% lower than that of wild type, indicating a defect in ABA biosynthesis. vp14 embryos were not deficient in epoxy-carotenoids, and extracts of vp14 embryos efficiently converted the carotenoid cleavage product, xanthoxin, to ABA, suggesting a lesion in the cleavage reaction. vp14 was cloned by transposon tagging. The VP14 protein sequence is similar to bacterial lignostilbene dioxygenases (LSD). LSD catalyzes a double-bond cleavage reaction that is closely analogous to the carotenoid cleavage reaction of ABA biosynthesis. Southern blots indicated a family of four to six related genes in maize. The Vp14 mRNA is expressed in embryos and roots and is strongly induced in leaves by water stress. A family of Vp14-related genes evidently controls the first committed step of ABA biosynthesis. These genes are likely to play a key role in the developmental and environmental control of ABA synthesis in plants. PMID- 9342394 TI - Conserved gene clusters in bacterial genomes provide further support for the primacy of RNA. AB - Five complete bacterial genome sequences have been released to the scientific community. These include four (eu)Bacteria, Haemophilus influenzae, Mycoplasma genitalium, M. pneumoniae, and Synechocystis PCC 6803, as well as one Archaeon, Methanococcus jannaschii. Features of organization shared by these genomes are likely to have arisen very early in the history of the bacteria and thus can be expected to provide further insight into the nature of early ancestors. Results of a genome comparison of these five organisms confirm earlier observations that gene order is remarkably unpreserved. There are, nevertheless, at least 16 clusters of two or more genes whose order remains the same among the four (eu)Bacteria and these are presumed to reflect conserved elements of coordinated gene expression that require gene proximity. Eight of these gene orders are essentially conserved in the Archaea as well. Many of these clusters are known to be regulated by RNA-level mechanisms in Escherichia coli, which supports the earlier suggestion that this type of regulation of gene expression may have arisen very early. We conclude that although the last common ancestor may have had a DNA genome, it likely was preceded by progenotes with an RNA genome. PMID- 9342395 TI - Rapid rate of control-region evolution in Pacific butterflyfishes (Chaetodontidae). AB - Sequence differences in the tRNA-proline (tRNApro) end of the mitochondrial control-region of three species of Pacific butterflyfishes accumulated 33-43 times more rapidly than did changes within the mitochondrial cytochrome b gene (cytb). Rapid evolution in this region was accompanied by strong transition/transversion bias and large variation in the probability of a DNA substitution among sites. These substitution constraints placed an absolute ceiling on the magnitude of sequence divergence that could be detected between individuals. This divergence "ceiling" was reached rapidly and led to a decay in the relative rate of control-region/cytb b evolution. A high rate of evolution in this section of the control-region of butterflyfishes stands in marked contrast to the patterns reported in some other fish lineages. Although the mechanism underlying rate variation remains unclear, all taxa with rapid evolution in the 5'-end of the control-region showed extreme transition biases. By contrast, in taxa with slower control-region evolution, transitions accumulated at nearly the same rate as transversions. More information is needed to understand the relationship between nucleotide bias and the rate of evolution in the 5'-end of the control-region. Despite strong constraints on sequence change, phylogenetic information was preserved in the group of recently differentiated species and supported the clustering of sequences into three major mtDNA groupings. Within these groups, very similar control-region sequences were widely distributed across the Pacific Ocean and were shared between recognized species, indicating a lack of mitochondrial sequence monophyly among species. PMID- 9342396 TI - Positional preferences of polypurine/polypyrimidine tracts in Saccharomyces cerevisiae genome: implications for cis regulation of gene expression. AB - The complete genome of the baker's yeast S. cerevisiae was analyzed for the presence of polypurine/polypyrimidine (poly[pu/py]) repeats and their occurrences were classified on the basis of their location within and outside open reading frames (ORFs). The analysis reveals that such sequence motifs are present abundantly both in coding as well as noncoding regions. Clear positional preferences are seen when these tracts occur in noncoding regions. These motifs appear to occur predominantly at a unit nucleosomal length both upstream and downstream of ORFs. Moreover, there is a biased distribution of polypurines in the coding strands when these motifs occur within open reading frames. The significance of the biased distribution is discussed with reference to the occurrence of these motifs in other known mRNA sequences and expressed sequence tags. A model for cis regulation of gene expression is proposed based on the ability of these motifs to form an intermolecular triple helix structure when present within the coding region and/or to modulate nucleosome positioning via enhanced histone affinity when present outside coding regions. PMID- 9342397 TI - Evolution of orthologous intronless and intron-bearing globin genes in two insect species. AB - While globin genes ctt-2beta and ctt-9.1 in Chironomus thummi thummi each have a single intron, all of the other insect globin genes reported so far are intronless. We analyzed four globin genes linked to the two intron-bearing genes in C. th. thummi. Three have a single intron at the same position as ctt-2beta and ctt-9.1; the fourth is intronless and lies between intron bearing genes. Finally, in addition to its intron, one gene (ctt-13RT) was recently interrupted by retrotransposition. Phylogenetic analyses show that the six genes in C. th. thummi share common ancestry with five globin genes in the distantly related species C. tentans, and that a 5-gene ancestral cluster predates the divergence of the two species. One gene in the ancestral cluster gave rise to ctn-ORFB in C. tentans, and duplicated in C. th. thummi to create ctt-11 and ctt-12. From parsimonious calculations of evolutionary distances since speciation, ctt-11, ctt 12, and ctn-ORFB evolved rapidly, while ctn-ORFE in C. tentans evolved slowly compared to other globin genes in the clusters. While these four globins are under selective pressure, we suggest that most chironomid globin genes were not selected for their unique function. Instead, we propose that high gene copy number itself was selected because conditions favored organisms that could synthesize more hemoglobin. High gene copy number selection to produce more of a useful product may be the basis of forming multigene families, all of whose members initially accumulate neutral substitutions while retaining essential function. Maintenance of a large family of globin genes not only ensured high levels of hemoglobin production, but may have facilitated the extensive divergence of chironomids into as many as 5000 species. PMID- 9342399 TI - Codon usage bias and tRNA abundance in Drosophila. AB - Codon usage bias of 1,117 Drosophila melanogaster genes, as well as fewer D. pseudoobscura and D. virilis genes, was examined from the perspective of relative abundance of isoaccepting tRNAs and their changes during development. We found that each amino acid contributes about equally and highly significantly to overall codon usage bias, with the exception of Asp which had very low contribution to overall bias. Asp was also the only amino acid that did not show a clear preference for one of its synonymous codons. Synonymous codon usage in Drosophila was consistent with "optimal" codons deduced from the isoaccepting tRNA availability. Interestingly, amino acids whose major isoaccepting tRNAs change during development did not show as strong bias as those with developmentally unchanged tRNA pools. Asp is the only amino acid for which the major isoaccepting tRNAs change between larval and adult stages. We conclude that synonymous codon usage in Drosophila is well explained by tRNA availability and is probably influenced by developmental changes in relative abundance. PMID- 9342400 TI - The molecular evolution of visual pigments of freshwater crayfishes (Decapoda: Cambaridae). AB - This study examines the diverse maximum wavelength absorption (lambdamax) found in crayfishes (Decapoda: Cambaridae and Parastacidae) and the associated genetic variation in their opsin locus. We measured the wavelength absorption in the photoreceptors of six species that inhabit environments of different light intensities (i.e., burrows, streams, standing waters, and subterranean waters). Our results indicate that there is relatively little variation in lambdamax (522 530 nm) among species from different genera and families. The existing variation did not correlate with the habitat differences of the crayfishes studied. We simultaneously sequenced the rhodopsin gene to identify the amino acid replacements that affect shifts in maximum wavelength absorption. We then related these to changes that correlated with shifts in lambdamax by reconstructing ancestral character states using a maximum-likelihood approach. Using amino acid sequences obtained from five species (all were 301 amino acids in length), we identified a number of candidates for producing shifts of 4 to 8 nm in lambdamax. These amino acid replacements occurred in similar regions to those involved in spectral shifts in vertebrates. PMID- 9342401 TI - Molecular evolution of the photolyase-blue-light photoreceptor family. AB - The photolyase-blue-light photoreceptor family is composed of cyclobutane pyrimidine dimer (CPD) photolyases, (6-4) photolyases, and blue-light photoreceptors. CPD photolyase and (6-4) photolyase are involved in photoreactivation for CPD and (6-4) photoproducts, respectively. CPD photolyase is classified into two subclasses, class I and II, based on amino acid sequence similarity. Blue-light photoreceptors are essential light detectors for the early development of plants. The amino acid sequence of the receptor is similar to those of the photolyases, although the receptor does not show the activity of photoreactivation. To investigate the functional divergence of the family, the amino acid sequences of the proteins were aligned. The alignment suggested that the recognition mechanisms of the cofactors and the substrate of class I CPD photolyases (class I photolyases) are different from those of class II CPD photolyases (class II photolyases). We reconstructed the phylogenetic trees based on the alignment by the NJ method and the ML method. The phylogenetic analysis suggested that the ancestral gene of the family had encoded CPD photolyase and that the gene duplication of the ancestral proteins had occurred at least eight times before the divergence between eubacteria and eukaryotes. PMID- 9342402 TI - The evolution of the conserved ATPase domain (CAD): reconstructing the history of an ancient protein module. AB - The AAA proteins (ATPases Associated with a variety of cellular Activities) are found in eubacterial, archaebacterial, and eukaryotic species and participate in a large number of cellular processes, including protein degradation, vesicle fusion, cell cycle control, and cellular secretory processes. The AAA proteins are characterized by the presence of a 230 to 250-amino acid ATPase domain referred to as the Conserved ATPase Domain or CAD. Phylogenetic analysis of 133 CAD sequences from 38 species reveal that AAA CADs are organized into discrete groups that are related not only in structure but in cellular function. Evolutionary analyses also indicate that the CAD was present in the last common ancestor of eubacteria, archaebacteria, and eukaryotes. The eubacterial CADs are found in metalloproteases, while CAD-containing proteins in the archaebacterial and eukaryotic lineages appear to have diversified by a series of gene duplication events that lead to the establishment of different functional AAA proteins, including proteasomal regulatory, NSF/Sec, and Pas proteins. The phylogeny of the CADs provides the basis for establishing the patterns of evolutionary change that characterize the AAA proteins. PMID- 9342403 TI - Tail-to-head arrangement of a partial chicken glyceraldehyde-3-phosphate dehydrogenase processed pseudogene. AB - A chicken glyceraldehyde 3-phosphate dehydrogenase (GAPDH) processed pseudogene was identified by inverse PCR using oligonucleotide primers specific for the 5' region of the GAPDH mRNA. Molecular cloning and sequence analysis of this genomic sequence shows that the processed pseudogene is incomplete and arranged in a permuted tail-to-head order. We propose that the tail-to-head organization is the result of circularization and breakage of a GAPDH retrogene prior to chromosomal integration. PCR analysis of DNAs from quail, pheasant, and various jungle fowl, shows that the processed pseudogene was formed after the three genera diverged but prior to Gallus speciation. This is the first report of a chicken GAPDH processed pseudogene sequence. This is also the first published report of a processed pseudogene with a tail-to-head organization. PMID- 9342404 TI - Chromosomal reorganization during meiosis of Saccharomyces cerevisiae baker's yeasts. AB - The genomic constitution of two S. cerevisiae baker's yeasts and their meiotic products have been analyzed by pulsed-field gel-electrophoresis, hybridization with specific gene probes, marker segregation, and flow cytometry. The parental strains have chromosomal patterns substantially different from those of laboratory strains used as controls. This pattern is partly the result of there being more than one copy of homologous chromosomes of different size, as judged by Southern-blot hybridization carried out with specific gene probes. Flow cytometry indicated that the strains have a 2.7 C DNA content. Tetrad analysis showed disomy for some chromosomes and tetrasomy for others. When two complete tetrads were subjected to molecular analysis the results confirmed instances of segregation of homologous chromosomes of different size. However, the presence of chromosomal bands absent in the parentals and the disappearance of chromosomal bands present in the parental strains were frequently seen. This result was attributed to two different phenomena: (1) the presence of multiple Ty1 and Ty2 transposable elements which seem to undergo interchromosomal translocation together with amplification, giving rise to differences in chromosomal size; (2) the presence of multiple Y' subtelomeric regions, giving rise to asymmetrical homologous recombination and, as a consequence, differences between the size of the recombinant chromosomes and the non-recombinant parental chromosomes. Chromosomal reorganization occurs with a very high frequency during meiosis. By contrast, mitosis is very stable, as judged by the reproducible electrophoretic karyotype shown by the parental strains in successive generations. PMID- 9342405 TI - MSS11, a novel yeast gene involved in the regulation of starch metabolism. AB - Expression of the STA1-3 glucoamylase genes, responsible for starch degradation in Saccharomyces cerevisiae, is down regulated by the presence of STA10. In order to elucidate the role of STA10 in the regulation of the glucoamylase system, a multicopy genomic library was constructed and screened for genes that enhanced growth of a STA2-STA10 S. cerevisiae strain on starch media. This screen allowed us to clone and characterize a novel activator gene of STA2 (and by extrapolation, STA1 and STA3), designated MSS11. A strain transformed with multiple copies of MSS11 exhibits increased levels of STA2 mRNA and, consequently, increased glucoamylase activity. Deletion of MSS11, located on chromosome XIII, results in media-dependent absence of glucoamylase synthesis. MSS11 has not been cloned previously and the encoded protein, Mss11p, is not homologous to any other known protein. An outstanding feature of Mss11p is that the protein contains regions of 33 asparagine residues interrupted by only three serine residues, and 35 glutamine residues interrupted by a single histidine residue. Epistasis studies showed that deletion of MSS11 abolishes the activation of STA2 caused by the over-expression of MSS10, a previously identified gene. In turn, it was found that deletion of MSS10 still allows activation of STA2 by over expression of MSS11. Mss11p therefore appears to be positioned below Mss10p in a signal transduction pathway. PMID- 9342406 TI - Cloning and characterization of KlCOX18, a gene required for activity of cytochrome oxidase in Kluyveromyces lactis. AB - We describe the isolation and initial characterization of KlCOX18, a gene that is essential for the assembly of a functional cytochrome oxidase in the yeast Kluyveromyces lactis. Cells carrying a recessive nuclear mutation in this gene are respiratory deficient and contain reduced levels of cytochromes a and a3. The KlCOX18 gene has been cloned by complementation of the respective nuclear mutation, sequenced, and disrupted. KlCOX18 is located on chromosome II and contains an open reading frame of 939 base pairs. The corresponding protein exhibits 70.4% similarity to the Cox18p of Saccharomyces cerevisiae. It contains three possible membrane-spanning domains and a putative amino-terminal mitochondrial import sequence. The strain carrying a null mutation in KlCOX18 does not grow on non-fermentable carbon sources and is deficient in both cytochrome c oxidase and respiratory activity. It is proposed that KlCox18p, like its S. cerevisiae counterpart, provides an important function at a later step of the cytochrome oxidase assembly pathway. PMID- 9342407 TI - The Neurospora crassa cya-5 nuclear gene encodes a protein with a region of homology to the Saccharomyces cerevisiae PET309 protein and is required in a post transcriptional step for the expression of the mitochondrially encoded COXI protein. AB - The cya-5 nuclear mutant of Neurosopora crassa was previously shown to be deficient in cytochrome aa3, cytochrome c oxidase activity, and the immunologically detectable COXI protein. We have now demonstrated that the mitochondria of this mutant contain mRNA for the COXI protein and that COXI cannot be detected during pulse-chase labeling experiments of mitochondrial translation products. Cloning and analysis of the cya-5 gene reveal a long open reading frame capable of encoding a 1136 amino-acid protein. Sequence analysis suggests that the potential CYA-5 protein contains a mitochondrial targeting sequence at its amino-terminus. The long open reading frame also contains a 200 amino-acid region with homology to the PET309 protein, which is required for the production or stability of intron-containing coxI mRNAs, as well as the translation of mature coxI mRNAs, in the yeast Saccharomyces cerevisiae. These data suggest that the CYA-5 protein of N. crassa is required in a post transcriptional step for COXI expression, most probably for the efficient translation of coxI mRNA. PMID- 9342408 TI - Evaluation of the efficiency of sexual reproduction in restoring Podospora anserina mitochondrial DNA to wild-type. AB - A Podospora anserina mitochondrial DNA (mtDNA) rearrangement mutant, Mn19, was crossed with a deletion mutant, alphaDelta5. Ascospores (212) from random asci were tested for viability, growth and life-span phenotypes, and mtDNA inheritance. Some spore inviability was detected along with early growth arrest (at the time of spore germination) from which some isolates recovered. However, the majority had wild-type growth and life-span phenotypes. All isolates tested at the DNA level (102) had wild-type mtDNA hybridization patterns with probes that detected defects in the parents. About 20% also inherited low levels of mtDNA molecules with the rearrangement characteristic of the Mn19 parent. These results demonstrate that P. anserina has a remarkable ability, through sexual reproduction, to restore its mtDNA to wild-type, even when the parents are predominately mutant. PMID- 9342409 TI - Transcriptional initiation sites in sorghum mitochondrial DNA indicate conserved and variable features. AB - Transcriptional initiation and processing was examined for three sorghum mitochondrial DNA genes (atp6-1, atp6-2, urf209) and two open reading frames (orf265/130, orf107) to characterize sequences associated with initiation and other transcriptional strategies for this species. The 5' termini of ten transcripts were determined by primer extension, and mtRNA was capped with guanylyl transferase and annealed to anti-sense riboprobes to identify transcriptional initiation regions. Eight transcript termini were suitable substrates for guanylyl transferase, indicating the presence of one (atp6-1, atp6 2, urf209), two (orf265/130), or three (orf107) promoters for the five examples. The majority of the putative promoters were associated with single primer extension termini, while two examples exhibited two transcript-initiation sites within the promoter. Four examples were characterized by initiated transcripts without subsequent processing, indicating that processing is not obligatory. Each of the putative promoter regions included significant A/T-rich 5' regions, consistent with previous examples, but four exceptions to a consensus core YRTA sequence were identified. The anomalies (AATA, CTTA) suggest plasticity in the primary structure of the core region of higher-plant mitochondrial DNA promoters. PMID- 9342410 TI - A deviant mitochondrial genetic code in prymnesiophytes (yellow-algae): UGA codon for tryptophan. AB - The sequence of a representative mitochondrial gene COXI, encoding cytochrome c oxidase subunit I, was determined in five species that cover all the orders of the Prymnesiophyta with the exception of the Pavlovales. Through this analysis, we noticed that the 'stop' codon UGA appears frequently and, specifically, at conserved tryptophan (Trp) sites of the gene. We showed these sites were not edited in the corresponding mRNA in one of these species, Isochrysis galbana. Therefore, it is most likely that the UGA codon is used for Trp, and not as a stop codon, in prymnesiophytes. All the analyzed prymnesiophytes made a tight cluster on the COXI phylogenetic tree which includes representative species of green-algae, land plants, yellow-green algae, eustigmatophytes and a red-alga. This suggests a monophyletic origin for the prymnesiophytes. The same deviant genetic code, i.e. UGA for Trp, has also been found in the red-alga, Chondrus crispus. In spite of the fact that this red-alga and the prymnesiophytes, share the same deviant genetic code for Trp, close affinity between the two groups was not statistically supported by the phylogenetic analysis of COXI sequences. PMID- 9342411 TI - Phylogeny and expression of the secA gene from a chromophytic alga--implications for the evolution of plastids and sec-dependent protein translocation. AB - In bacteria many periplasmatic proteins are exported via the sec-dependent pathway. A homologous apparatus was found to be involved in the transport of proteins across the thylakoid membrane in plastids. In the present study additional data on the phylogeny and expression of one of the genes essential in this process, secA, is presented. For the first time, transcriptional activity of secA in the plastid was detected. When secA is used as a phylogenetic marker for plastid evolution it demonstrates a large phylogenetic distance between chlorophytic and non-chlorophytic (i.e. rhodophytic) primary plastids. This distance could be explained by assuming polyphyly for major plastid lineages. Moreover, it was found that two types of secA genes may exist in plastids. Whether or not these are involved in different protein translocation processes is presently unknown. In an attempt to identify further candidates, i.e. non photosynthesis-related proteins, for sec-dependent protein transport, an SbpA protein was detected in chromophytic plastids by the use of a peptide antibody. PMID- 9342412 TI - Synthesis and evaluation of 2-chloromethyl-3-N-substituted arylthieno (2,3 d)pyrimidin-4-ones and derivatives for central nervous system depressant activity. AB - 2-Chloromethyl 3-N-substituted arylthieno (2,3-d)pyrimidin-4-ones and derivatives were synthesized by reacting 2-amino-3-carboxanilido-4,5,6,7 tetrahydrobenzo(b)thiophenes (Iabc) with chloroacetyl chloride in dioxane and by cyclizing the open chain 3-substituted carboxanilido-2-(omega-chloroacetamido) 4,5,6,7-tetrahydrobe nzo(b)thiophenes (IIabc) under acidic condition. The compounds were characterized by their spectral data and screened for central nervous system depressant activity. The compounds IIIabc and Vabc have shown marked sedative action. PMID- 9342413 TI - Role of I1 imidazoline receptors in the sympathoinhibition produced by intracisternally administered rilmenidine and moxonidine. AB - The role of alpha 2-adrenoceptors and I1 imidazoline receptors in the cardiovascular effects of rilmenidine (CAS 54187-04-1), moxonidine (CAS 75438-57 2) and guanabenz (CAS 23256-50-0) was studied in conscious rabbits. In radioligand binding studies, rilmenidine and moxonidine are selective for I1 imidazoline receptors (vs. alpha 2-adrenoceptors) and guanabenz is selective for alpha 2-adrenoceptors (vs. I1 imidazoline receptors). Efaroxan (CAS 89197-00-2; selective for I1 imidazoline receptors) and yohimbine (CAS 65-19-0; selective for alpha 2-adrenoceptors) were used as antagonists. All drugs were injected into the cisterna magna. Guanabenz (1, 3, 10 and 30 micrograms kg-1), rilmenidine (1, 3, 10 and 30 micrograms kg-1) and moxonidine (0.03, 0.1, 0.3 and 1 microgram kg-1) dose-dependently lowered blood pressure, heart rate and the plasma concentration of noradrenaline. In the antagonism experiments, guanabenz (10 micrograms kg-1), rilmenidine (10 micrograms kg-1) and moxonidine (0.3 micrograms kg-1) were administered first; they caused equal hypotension. Injection of the agonists was followed by injection of efaroxan (0.3-1 microgram kg-1) or yohimbine (0.1-0.3 micrograms kg-1). Efaroxan and yohimbine were equieffective at antagonizing the effects of guanabenz. In contrast, efaroxan was more effective than yohimbine at antagonizing the effects of rilmenidine and moxonidine. The results show that intracisternally administered guanabenz, rilmenidine and moxonidine cause sympathoinhibition. alpha 2-Adrenoceptors are responsible for the sympathoinhibition produced by guanabenz. In contrast, I1 imidazoline receptors are involved in the sympathoinhibition caused by rilmenidine and moxonidine. PMID- 9342415 TI - Anticoagulant activity of the novel thrombin inhibitor 1-butyl-3-(6,7-dimethoxy-2 naphthylsulfonyl) amino-3-(3-guanidinopropyl)-2-pyrrolidinone hydrochloride. AB - 1-Butyl-3-(6,7-dimethoxy-2-naphthylsulfonyl)amino-3-(3-guanidin opropyl)-2 pyrrolidinone hydrochloride (CAS 173440-64-7, SPI-501), which has highly selective thrombin-inhibitory activity, caused a concentration-dependent increase in the time taken for coagulation induced by thrombin in rabbit plasma. The IC50 of SPI-501 was 1.7 mumol/l. Argipidine also prolonged coagulation time and its activity was one order of magnitude greater than that of SPI-501. SPI-501 and argipidine caused dose-dependent increases in the activated partial prothrombin time (APTT) and prothrombin time (PT) of rat plasma. When APTT and PT were measured, IC50 values of SPI-501 were 38.0 and 18.5 mumol/l and those of argipidine, 1.4 and 1.9 mumol/l, respectively. Intravenous administration of SPI 501 (10 and 30 mumol/kg) and argipidine (1 and 3 mumol/kg) prolonged both APTT and PT in rats. While SPI-501 was less potent than argipidine, the durations of the effects of both were the same. PMID- 9342414 TI - Long-term blockade of the angiotensin II receptor in renin transgenic rats, salt loaded Dahl rats, and stroke-prone spontaneously hypertensive rats. AB - These studies were designed to investigate the protective effects of the new angiotensin II receptors antagonist BAY 10-6734 (6-n-butyl-4-methoxycarbonyl-2 oxo-1[(2'-(1H-tetrazol-5-yl) -3-fluorobiphenyl-4-yl)methyl] 1,2-dihydropyridine, CAS 156001-18-2) on haemodynamic, hormonal, renal, and structural parameters in renin transgenic rats (TGR(mRen2)27), salt-loaded Dahl S and R rats, and salt loaded stroke-prone spontaneously hypertensive rats (SHR-SP) in long-term trials. Study 1: In SHR-SP the development of blood pressure, cardiac hypertrophy, and the deleterious effects of salt loading on kidney structure and kidney function was prevented by BAY 10-6734. Study 2: In salt-loaded Dahl S rats with a suppressed plasma renin activity treatment with BAY 10-6734 did not delay the increase in blood pressure but prevented cardiac hypertrophy and the increase in plasma ANP (Atrial natriuretic peptide). Study 3: TGR develop malignant hypertension associated with cardiac hypertrophy, elevated left-ventricular end diastolic pressure and increased plasma ANP. After 6 weeks of treatment with BAY 10-6734 (30 mg/kg p.o. bid) cardiac pump function was improved and cardiac hypertrophy was reversed in this angiotension dependent form of hypertension. The beneficial effects of BAY 10-6734 in these different animal hypertension models are also emphasized by a reduction in mortality. PMID- 9342417 TI - Evaluation of phototoxic properties of oral antidiabetics and diuretics. Photohemolysis model as a screening method for recognizing potential photosensitizing drugs. AB - The oral hypoglycemic drugs carbutamide, chlorpropamide, glibenclamide, glubornuride, gliclazide, glipizide, gliquidone, glisoxepide, glymidine, tolazamide and tolbutamide, and the diuretics acetazolamide, bemetizide, bendroflumethiazide, benzthiazide, benzylhydrochlorothiazide, bumetanide, butizide, chlorazanile, chlorothiazide, chlortalidone, clopamide, cyclopenthiazide, cyclothiazide, diazoxide, etozoline, furosemide, hydrochlorothiazide, hydroflumethiazide, indapamide, mefruside, metolazone, piretanide, polythiazide, trichlormethiazide, and xipamide were investigated for photohemolytic properties in vitro. Irradiation with a SOL 3 apparatus (solar simulating irradiation) revealed hemolysis in the presence of five oral hypoglycemic agents and in the presence of 19 out of the 25 tested diuretics. Photohemolysis was induced in the presence of three substances, respectively, after exposure to UVA or visible light. UVB alone did not induce phototoxic hemolysis in the presence of the tested drugs. Compared to clinical reports on photosensitivity reactions, the photohemolysis model seems a good predictive model in recognizing potential photosensitizing sulfonamides. PMID- 9342416 TI - Effects of the novel thromboxane (TXA2) receptor antagonist linotroban on inulin and para-aminohippuric acid clearances in the conscious male and female rat. AB - This paper reports the results of experiments designed to determine the effect of linotroban (CAS 141443-73-4) a selective thromboxane (TXA2) receptor antagonist with novel antithrombotic activity, on renal function in conscious male and female rats. Linotroban was administered subcutaneously at doses of 0, 6, 24, 48 and 96 mg/kg/24 h by osmotic minipumps over 6 days. Inulin (Inutest) and para aminohippuric acid (PAH) clearances were determined on the last day of linotroban treatment. These agents were delivered by a slow release tablet planted s.c. five days after the start of the linotroban treatment. Linotroban did not significantly alter renal functions, although there was a significant difference in GFR (glomerular filtration rate) for male and female rats receiving the highest dose. Linotroban is well tolerated but the difference in renal function between male and female rats at the highest dose may reflect a threshold of renal tolerance. PMID- 9342418 TI - Response of human ciliated respiratory cells to a mixture of menthol, eucalyptus oil and pine needle oil. AB - Nasal respiratory cells were harvested from 45 healthy individuals using a microcurette technique. Following harvest, cells were placed on microporous polycarbonate membranes and exposed for 30 min using a gas/liquid interface technique to a mixture of menthol, eucalyptus oil and pine needle oil (MEP), eucalyptus oil alone (ECO), and pine needle oil alone (PNO) at concentrations ranging between 0.2 and 11 g/m3. Ambient air served as control. Ciliary beat frequency (CBF) was assessed before and after exposure using video-interference contrast microscopy. Control exposure to air (n = 10) did not alter CBF (7.1 +/- 0.9 Hz before and 7.0 +/- 0.9 Hz after exposure, -1.3% decrease), whereas a dose dependent decrease of CBF following exposure to MEP (max. decrease -22.6% at a concentration of 10 g/m3, p < 0.01), ECO (max. decrease -32.5% at a concentration of 7.5 g/m3, p < 0.01) and PNO (max. decrease -56.1% at a concentration of 9.4 g/m3, p < 0.01) was observed. The data show that essential oils in high concentrations can reduce in-vitro ciliary activity of human respiratory cells not protected by a mucus layer. These effects have to be verified by in vivo investigations. PMID- 9342419 TI - Determination of diclofenac in micro-whole blood samples by high-performance liquid chromatography with electrochemical detection. Application in a pharmacokinetic study. AB - A rapid and sensitive method for the determination of diclofenac (CAS 15307-86-5) in whole blood samples by high-performance liquid chromatography with amperometric detection has been developed. This method was then used to study the pharmacokinetics of oral diclofenac sodium in the rat. The method includes a single extraction of acidified whole blood with ethyl acetate. Extracts were analyzed on a reversed-phase column eluted with a mixture of acetonitrile and 0.075 mol/l sodium acetate solution (pH 3.3) and detected amperometrically at + 1.1 V against Ag/AgCl. Retention times for diclofenac and the internal standard (naproxen) were 3.5 and 6 min, respectively. The method was linear in the range of 25 to 2000 ng/ml and the detection limit of the method was 10 ng/ml, using 100 microliters of whole blood sample. Employing this method, the oral pharmacokinetics of diclofenac in the rat was studied. Wistar male rats received an oral dose of 1, 3.2 or 10 mg/kg of diclofenac and blood samples were drawn at selected times during 12 h. After administration of diclofenac, a rapid increase of circulating concentrations was observed reaching a maximum in about 10 min. Then concentration decayed with a half-life of about 15 h. It is concluded that the method here reported is adequate for realization of pharmacokinetic studies of diclofenac in small species. PMID- 9342420 TI - Determination of a new hypoxia selective agent from 2-quinoxalinecarbonitrile 1,4 di-N-oxides in plasma by high performance liquid chromatography. AB - A high performance liquid chromatography (HPLC) method was developed and validated for the determination of 7-chloro-3-[[(N,N-dimethylamino)propyl]amino] 2-quinaxolinecarbonitri le 1,4-di-N-oxide (Q-85), a new hypoxia-selective agent, in plasma. The assay involves extraction into diethylether-dichloromethane (2:1) from plasma and subsequent analysis by reversed-phase HPLC with ultraviolet detection at 292 nm. Response was linear in the range of 0.4-20 micrograms/ml of plasma and the detection limit was 0.05 micrograms/ml. The extraction recovery was about 90%. The within and between day coefficients of variation did not exceed 6.1%. This HPLC assay has been applied to determine Q-85 in plasma samples taken during pharmacokinetic studies in mice. PMID- 9342421 TI - Toxicological profile of a low molecular weight spleen peptide formulation used in supportive cancer therapy. AB - The toxicity of Polyerga (GP-1), a mixture of low molecular weight spleen peptides, extracted from porcine spleen was examined in in vitro and in vivo experiments. The following endpoints were examined: single dose toxicity, repeated dose toxicity, reproduction toxicity, mutagenic potential, local tolerance, immunotoxicity and general pharmacology. The standard therapeutic dose is 3 x 1 ml GP-1/week. The dosage can be increased temporarily to 2 ml/d. 1 ml injection solution contains 30 micrograms oligopeptides. The LD50 was determined in rats as 3.76 ml/kg b.w. i.v. Following repeated intramuscular administration of GP-1 no toxicity appeared in rats up to a dose level of 2 ml/kg b.w./d during the 13-week treatment period. In dogs the no-effect level of GP-1 was 0.32 ml/kg b.w./d i.m. (marginal body weight reduction at 0.80 ml/kg b.w./d i.m.). In an embryotoxicity study in rats, GP-1 possessed no teratogenic properties tested at dose levels exceeding the human therapeutic dose by a factor of 1000. No mutagenic potential was observed for GP-1 in the AMES test. No local irritations were observed at the intended injection sites and at injection sites made in error. None of the in vitro and in vivo pharmacological studies revealed any effect on the parameters tested, at doses up to 500 times the clinical dose. Hence, under the present test conditions, based on a daily therapeutic dose of 1 ml GP-1/patient (approximately 0.014 ml/kg b.w./d), the therapeutic ratio is at least 50 for the most sensitive endpoint 'repeated dose toxicity' reflecting the wide margin of safety for GP-1. PMID- 9342422 TI - Pharmacodynamic parameters of ofloxacin, tobramycin and ceftriaxone and their effect on the biological properties of salmonellae. AB - The postantibiotic effect and postantibiotic sub-MIC effect of ofloxacin (CAS 82419-36-1), tobramycin (CAS 32986-56-4) and ceftriaxone (CAS 73384-59-5) on two salmonella serotypes (S. typhimurium and S. enteritidis) were studied. The influence of postantibiotic effect and postantibiotic sub-MIC effect of the antibiotics on prophage induction of the lysogenic S. typhimurium strain and on Congo red binding by both serovars as indicator of their invasive ability was examined. The postantibiotic effect was induced by exposure of the bacteria to the 2x and 4x MIC concentrations of antibiotics studied for 0.5 h. The postantibiotic effects were different; ceftriaxone induced the longest postantibiotic effect against S. enteritidis, and the 4x MIC of tobramycin induced the longest postantibiotic effect against S. typhimurium. The postantibiotic sub-MIC effects lasted longer and in the case of subinhibitory concentrations of tobramycin on S. typhimurium and ceftriaxone on S. enteritidis did not allow any regrowth. The results showed that the postantibiotic effect and postantibiotic sub-MIC effect of ofloxacin induced a prophage of a lysogenic S. typhimurium strain, and the postantibiotic sub-MIC effects of tobramycin influenced Congo red binding by S. enteritidis cells. PMID- 9342423 TI - Studies on the antifungal activity of the new imidazole antimycotic lanoconazole in infected sites. Distribution in the skin and in vitro activity in the presence of stratum corneum. AB - To answer the question why topically applied lanoconazole (CAS 101530-10-3, NND 318) is so highly effective in the treatment of dermatomycoses in both animal models and human patients, the antifungal activity of lanoconazole in infected sites was investigated. 1. Distribution of lanoconazole in rat skin: The distribution of lanoconazole within the dorsal skin of rats was examined by measurement of radioactivity and microscopic autoradiography. 24 h after dermal application of 14C-lanoconazole cream formulation, 83% of the total radioactivity in the skin was recovered from the stratum corneum, and thereafter the radioactivity decreased gradually up to 96 h. Metabolite analysis showed that more than 94% of the extractable radioactivity was lanoconazole itself after 24 and 48 h. Microautoradiograms of the skin also supported the radioactivity distribution. 2. In vitro antifungal activity in stratum corneum-containing medium: Candida albicans TIMM 2640 was incubated for 10 days at 27 degrees C in a vitamin-supplemented yeast carbon base medium containing 5 mg/ml of human stratum corneum and different concentrations of lanoconazole or bifonazole (CAS 60628-96 8, reference agent). Compared with the control culture, lanoconazole strongly inhibited fungal growth in a concentration dependent manner at concentration above 0.1 microgram/ml, resulting in a reduction of viable cell recovery to less than 50% after 10 days. The inhibitory activity of bifonazole was clearly weaker than that of lanoconazole. At concentrations of 0.1-10 micrograms/ml lanoconazole reduced keratinolytic acid proteinase activity in the culture-supernate to 40-70% of the control value, while bifonazole showed 50% reduction of the activity at a concentration of 10 micrograms/ml. These results indicate that lanoconazole is mainly distributed and retained in the stratum corneum after topical application where it exerts strong antifungal activity. PMID- 9342424 TI - Pseudomonas aeruginosa infection in embryonated hen's eggs. An alternative in vivo model for the screening of antibacterial substances. AB - Embryonated hens' eggs can be reliably infected by Pseudomonas aeruginosa in laboratory experiments. Therapy tests with the antibiotics azlocillin (CAS 37091 66-0) and gentamicin (CAS 13291-74-2) on this type of infected hens' eggs demonstrate that this test system offers a realistic alternative to septic experiments with small laboratory rodents. Chick embryos survive a lethal Pseudomonas infection when azlocillin or gentamicin in a relevant therapeutic dose are administered immediately after the infective agent. The use of Pseudomonas infected chick embryos in the screening for new antiinfectives allows, therefore, a considerable reduction of the number of laboratory rodents required. PMID- 9342425 TI - Biological activity and therapeutic potential of homologs of an Ii peptide which regulates antigenic peptide binding to cell surface MHC class II molecules. AB - An invariant chain peptide (murine Ii76-92; Ii-Key) is known to produce a 10 to 50 times baseline enhancement of the presentation of specific antigenic peptides to murine T cell hybridomas by cell surface MHC class II molecules. In order to define structure-activity relationships in Ii-key, homologs were synthesized with the following systematic variations: 1) N- and C-terminal truncations, 2) N terminal acetylation and C-terminal amidation, 3) substitutions with 13 natural amino acids in each position of the shortest, fully active peptide, and then with an additional 12 nonnatural amino acids at certain 'pharmacophore' positions, 4) substitutions with D-amino acids and N-methyl-leucine, and 5) cyclical forms. More than 160 homologs were tested for effects on antigen presentation by the murine MHC class II alleles: A(d), Ak, E(d), or Ek. For some compounds, allele specificity between E(d) and Ek exceeded 1:20. D-Amino acid and/or N-methyl leucine substitutions were accepted at some residue positions, leading to peptides with relatively long half-lives in mouse serum and low toxicities in mice. An Ii-Key homolog inhibited in vitro presentation of an internally processed hen egg lysozyme determinant to a specific T hybridoma. The best compounds can be tested in vivo for therapeutic applications: 1) immunosuppression upon release of antigenic peptide, and 2) vaccination or immunomodulation upon co-administration of a second antigenic peptide. PMID- 9342426 TI - Micromet. PMID- 9342427 TI - The leukocyte cell adhesion cascade and its role in myocardial ischemia reperfusion injury. AB - Cell-cell and cell-matrix interactions are known to be mediated by specific cell adhesion receptors expressed on the cell surface. The characterization of these cell adhesion molecules has allowed researchers to examine their roles in a variety of physiologic and pathophysiologic conditions. Numerous studies have demonstrated that myocardial ischemia-reperfusion injury is an acute inflammatory process in which leukocytes are intimately involved. In this review, we summarize the current data on the leukocyte cell adhesion cascade, focus upon studies which have demonstrated specific cell adhesion molecule interactions which mediate the leukocyte involvement in myocardial ischemia-reperfusion injury and suggest future avenues of exploration and possible clinical implications of the studies reviewed. PMID- 9342428 TI - Changes in microsomal membrane phospholipids and fatty acids and in activities of membrane-bound enzyme in diabetic rat heart. AB - Diabetes mellitus is associated with alterations in lipid metabolism and cardiac dysfunction despite an absence of coronary arteriosclerotic changes. To investigate mechanisms of cardiac dysfunction in diabetic cardiomyopathy, we studied the relation between activities of membrane-bound enzymes and surrounding phospholipids in rats with diabetes induced with a single intravenous injection of streptozotocin (65 mg/kg). We found that total phospholipid content of sarcoplasmic reticulum membrane increased significantly 8 weeks after treatment with streptozotocin owing to increases in phosphatidylcholine and phosphatidylethanolamine, a decrease in arachidonic acid, and an increase in docosahexaenoic acid in the early stage of diabetes. Sarcolemmal Na+/K(+)-ATPase activity and the number of receptors decreased in isolated cardiomyocytes of diabetic rats 8 weeks after streptozotocin administration. The Ca2+ uptake of both sarcoplasmic reticulum and mitochondria decreased simultaneously in permeabilized, isolated cardiomyocytes from diabetic rats. The depression of membrane-bound enzyme activities was correlated with alterations in phospholipids, which are closely related to the microenvironment of membrane bound enzymes and influence intracellular Ca2+ metabolism. Because these changes in phospholipids and fatty acids were reversible with insulin therapy, they are diabetes-specific and might be a cause of cardiac dysfunction in diabetes. PMID- 9342429 TI - Prolongation of life span in hypertensive rats by dietary interventions. Effects of garlic and linseed oil. AB - The effects of long-term dietary application of garlic (dried powder, 0.5% in weight of standard chow; G group) or linseed oil (2.5%; L group) as well as a combination of both interventions (L + G group) on the life span of hypertensive rats (SHR SP) was investigated. A further group fed with standard chow served as control (C). The dietary interventions were started at the age of three weeks. Besides regular measurements of the systolic arterial blood pressure (oscillometrically at the tail artery) as well as of heart rate and body weight, autopsy and histological investigations were performed. Both diets, and particularly their combination, prolonged life span significantly (mean values (days) C: 434.5 +/- 23.5; G: 453.2 +/- 16.2; L: 470.0 +/- 26.2; L + G: 494.8 +/- 39.2). There was no significant interaction of the factors garlic and linseed oil. Systolic blood pressure as measured during the compensatory stage (data used until the 39th week of life) was significantly lowered by both garlic (mean -5.8 mm Hg), linseed oil (mean -6.3 mm Hg), and their combination (mean -11.3 mm Hg). The animals died as a consequence of congestive left and right ventricular failure with ventricular hypertrophy, dilatation, myocardial fibrosis and cellular infiltration, left ventricular atrial thrombosis (in most cases), and terminal pneumonia. On the other hand, arteriosclerotic plaques and signs of cerebral stroke could not be detected. Except for the degree of hypertrophy, which was lower in the treated groups, no differences were obvious regarding the morphological findings at the time of death. There was a significant positive correlation between mean blood pressure and the degree of left ventricular hypertrophy. Furthermore, a significant negative correlation between mean blood pressure and ventricular hypertrophy on the one hand and survival on the other hand was obvious provided the total number of animals was considered, however, not within the individual groups. The same applies to the relation between the reduction of left ventricular hypertrophy and life span. The relatively slight hypotensive effect of both dietary interventions as well as the results of previous investigations speaks in favor of a substantial influence of factors independent of blood pressure. In view of controversial results and interpretations in international literature, the mechanisms involved need further study. PMID- 9342430 TI - Ca(2+)-activated K+ channels in human smooth muscle cells of coronary atherosclerotic plaques and coronary media segments. AB - The behavior of Ca(2+)-activated K+ channels of large conductance (BKCa) in smooth muscle cells, which were obtained from atherosclerotic plaque material (SMCP) and from media segments (SMCM) of human coronary arteries, were compared using the patch-clamp technique. Voltage-clamp protocols in cell-attached patches revealed the characteristic voltage-dependent activation of BKCa in both cell groups. Single-channel conduction as 216.4 +/- 16.7 pS (n = 6) in SMCP and 199.9 +/- 6.7 pS (n = 6) in SMCM in symmetrical 140 mM K+ solutions. Using outside-out patches, external perfusion with 500 microM tetraethylammonium ions caused a typical "flickery block" of the unitary current. The selective BKCa channel inhibitor iberiotoxin (50 nM) effectively blocked BKCa, channel activity. Comparing BKCa open-state probabilities (P0) at +80 mV in cell-attached patches, a highly significant difference between SMCP (P0 = 0.1438 +/- 0.1301; n = 15) and SMCM (P0 = 0.0093 +/- 0.0044; n = 15; Kruskal-Wallis test, p < 0.001) was found. In contrast to this finding, there was no significant difference in the open state probability of BKCa, between SMCP (P0 = 0.542 +/- 0.0237; n = 9) and SMCM (P0 = 0.0472 +/- 0.0218; n = 10; p = n.s.) using inside-out patches. The results show an interesting difference in the behavior of large conductance Ca(2+) activated K+ channel in SMCP compared to SMCM with a significantly higher channel activity in human smooth muscle cells obtained from coronary atherosclerotic plaque material. This finding may indicate an important functional role of BKCa channels in the development of atherosclerosis. PMID- 9342431 TI - Effects of ischemia, preconditioning, and adenosine deaminase inhibition on interstitial adenosine levels and infarct size. AB - In order to examine the relationship between local adenosine concentrations before, during, and after ischemia and the extent of ischemic myocardial damage, measurements of interstitial fluid (ISF) nucleosides were made using microdialysis probes implanted in the ischemic region of isoflurane anesthetized Micropigs undergoing 60' coronary artery occlusion (CAO) and 3 h of reperfusion (REP). Nucleoside concentrations in the dialysate collected from the microdialysis probes were used as an index of ISF levels. Dialysate nucleoside concentrations (ADO, inosine and hypoxanthine), myocardial infarct size, and myocardial blood flow (MBF) were determined in control animals (n = 6), animals preconditioned with a single 10' cycle of CAO and REP (PC, n = 6), and those treated with the adenosine deaminase inhibitor pentostatin (n = 6, 0.2 mg/Kg i.v. 30' prior to CAO). The brief PC occlusion resulted in a transient but significant increase in dialysate ADO (6.7 +/- 1.8 microM vs. 0.67 +/- 0.1 microM at baseline). Pentostatin administration had no significant effect on either dialysate nucleosides or MBF at baseline. During the 60' CAO, dialysate ADO increased in control animals. In PC animals, however, dialysate ADO during CAO was lower than control. Pretreatment with pentostatin resulted in a six-fold augmentation in dialysate ADO during the 60 min CAO when compared to the control values (110.62 +/- 30.2 microM vs. 16.31 +/- 2.1 microM at 60 min of ischemia). Pentostatin also resulted in a significant reduction in the accumulation of inosine and hypoxanthine, indicating inhibition of adenosine deaminase activity. There were no significant differences in MBF between groups at any time point. Following 3 h REP, infarct size was 35.4 +/- 5.5%, 8.1 +/- 1.5% and 8.3 +/- 1.8% of the region at risk in control, PC, and pentostatin groups, respectively. These data suggest that marked increase in ISF ADO during CAO, may be as effective in reducing INF as a modest increase in ISF ADO prior to prolonged CAO. PMID- 9342432 TI - Ischaemic preconditioning may abolish the protection afforded by ATP-sensitive potassium channel openers in isolated human atrial muscle. AB - The ATP-sensitive potassium channel (KATP channel) has been implicated in the mechanism underlying ischaemic preconditioning protection. This study based on human atrium compared the protective effects of ischaemic preconditioning with pre-operative nicorandil (a KATP channel opener with nitrate actions). We also examined the added effect of ischaemic preconditioning to that of nicorandil on ischaemic protection. The protective effects of other KATP channel openers devoid of nitrate actions were also examined. Atrial trabeculae harvested from patients undergoing routine myocardial revascularisation were divided on the basis of whether patients had been ingesting nicorandil orally preoperatively. Trabeculae were superfused with oxygenated Tyrode's solution and following stabilisation underwent 90 minutes simulated ischaemia followed by 120 minutes reoxygenation (n = 6 per group). Atrial trabeculae exposed to nicorandil underwent either no treatment (N), or ischaemic preconditioning (N + PC) using 3 minutes simulated ischaemia and 7 minutes reoxygenation prior to the 90 minutes simulated ischaemia. Similarly trabeculae not exposed to nicorandil underwent either no treatment, controls (C), or ischaemic preconditioning (PC). The experimental endpoint was recovery of contractile function presented as percentage baseline function. Further groups were examined using other KATP channels openers with and without ischaemic preconditioning. In the control group, following 120 minutes reoxygentation the recovery of function reached 28.8 +/- 3.5%. In contrast, exposure to nicorandil alone improved recovery of function (55.5% +/- 5.3) to a similar extent as PC (55.3% +/- 2.5) when compared to controls (p < 0.05, ANOVA). The addition of ischaemic preconditioning to nicorandil exposure abolished protection (29.7% +/- 3.1 ). Findings were confirmed using the other KATP channels openers. Clinically available nicorandil appears to afford ischaemic protection to isolated human atrial muscle. The addition of a short ischaemic episode to nicorandil exposure seems to completely abolish this protection. Although the mechanism underlying this effect remains unknown, we believe that this observation may have clinical implications. PMID- 9342433 TI - Myocardial length-force relationship in end stage dilated cardiomyopathy and normal human myocardium: analysis of intact and skinned left ventricular trabeculae obtained during 11 heart transplantations. AB - The Frank-Starling-mechanism (FSM) was analyzed in isolated intact and skinned human left ventricular myocardium obtained from 11 heart transplantations (normal donor hearts (NDH), n = 8; dilated cardiomyopathy (DCM), n = 11). The new technique to utilize muscle strips from normal donor hearts which were actually implanted is described in detail. METHODS: I) In electrically stimulated left ventricular trabeculae (37 degrees C, oxygenated Krebs-Henseleit solution, supramaximal electrical stimulation, frequency 1 Hz) force development was analyzed as a function of muscle length (NDH = 8; DCM = 11). II) In an additional series left ventricular myocardium was demembranized ("skinned") by Triton-X-100. At different sarcomere lengths and calcium concentrations corresponding to pCa values of 4.3, 5.5, and 8.0 force development was measured (DCM = 11; NDH = 9). RESULTS: I) Developed force increased up to an optimum as a function of muscle length in intact NDH- and DCM-myocardium. However, the relative increment of developed force after any length step was smaller in DCM than in NDH. Near "Lmax" (muscle length associated with maximum developed force) passive resting tension was considerably elevated in DCM, indicating significantly increased diastolic stiffness. II) In skinned left ventricular DCM- and NDH-myocardium developed force depended on sarcomere length with an optimum near 2.2 microns. However, a reduction of activator calcium concentration from pCa 4.3 to pCa 5.5 produces a smaller percent decline in force at short sarcomere lengths in DCM than it does in NDH. CONCLUSION: The present study shows that except for diastolic stiffness and a smaller relative force increment after any length step in DCM the Frank Starling mechanism is still present in isolated human left ventricular DCM- as in NDH-myocardium. The current study does not allow to decide whether in skinned myocardium the smaller percent decline in force after reduction of activator calcium concentrations in DCM is caused by an increased calcium sensitivity at short sarcomere lengths or decreased sensitivity at long sarcomere lengths. PMID- 9342435 TI - Tamoxifen induced apoptosis in ZR-75 breast cancer xenografts antedates tumour regression. AB - The ZR-75-1 ER positive breast cancer cell line, xenografted in female nude mice, has been used to determine the effect of tamoxifen on cell proliferation (as measured by mitosis) and cell death (as evidenced by apoptosis and necrosis). After 2 days treatment, there was a significant rise in apoptosis (p < 0.05), whereas a fall in mitosis was not apparent until 7 days (p < 0.05). Furthermore there was an increase in the apoptotic:mitotic ratio on day 7 (p < 0.05). These changes antedated tumour regression, which did not reach not significance until day 14. Tamoxifen did not increase necrosis (which significantly decreased in treated tumours once they had regressed (p < 0.01). In contrast tamoxifen treatment of xenografted MDA-MB-231 ER-negative breast cancer cells produced no significant effects on growth, apoptosis, or mitosis. This study presents clear evidence for tamoxifen inducing apoptosis in ZR-75-1 xenografts (but not MDA-MB 231 tumours). Since changes in apoptosis and mitosis antedate tumour regression, their assessment may provide the potential by which to predict tumour response to tamoxifen therapy. PMID- 9342434 TI - Effect of chronotropic and inotropic stimulation on the coronary pressure-flow relation in left ventricular hypertrophy. AB - Left ventricular hypertrophy (LVH) secondary to chronic pressure overload is associated with increased susceptibility to myocardial hypoperfusion and ischemia during increased cardiac work. The present study was performed to study the effects of chronotropic and inotropic stimulation on the coronary pressure-flow relation of the hypertrophied left ventricle of dogs and to determine the individual contributions of increases in heart rate and contractility to the exaggerated exercise-induced increases in effective back pressure (pressure at zero flow; Pzf). Ascending aortic banding in seven dogs increased the LV to body weight ratio to 7.7 +/- 0.3 g/kg compared to 4.8 +/- 0.2 g/kg in 10 normal dogs (p < or = 0.01). Maximum coronary vasodilation was produced by intracoronary infusion of adenosine. During resting conditions maximum coronary blood flow in the pressure overloaded hypertrophied left ventricle was impaired by both an increase in Pzf (25.1 +/- 2.6 vs 13.8 +/- 1.2 mmHg in hypertrophied vs normal ventricles, respectively, p < or = 0.01) and a decrease in maximum coronary conductance (slope of the linear part of the pressure-flow relation, slopep > or = linear) (8.6 +/- 1.1 vs 12.7 +/- 0.9 ml/min/mmHg, p < or = 0.01). Right atrial pacing at 200 and 250 beats/min resulted in similar rightward shifts of the pressure-flow relation in hypertrophied and normal hearts with 3.1 +/- 0.8 and 4.7 +/- 0.8 mmHg increases in Pzf in LVH and normal dogs, respectively; stepwise multivariate regression analysis indicated that the exaggerated decrease in filling pressure (10 +/- 2 vs 6 +/-2 mmHg) and decrease in left ventricular systolic pressure (45 +/- 5 vs 3 +/- 3 mmHg, p < or = 0.01) may have blunted a greater rightward shift of the pressure-flow relation produced by atrial pacing in the hypertrophied hearts. Inotropic stimulation with dobutamine (10-20 micrograms/kg/min, i.v.) resulted in minimal flow changes in normal hearts but produced a 4.4 +/- 1.5 mmHg (p < or = 0.05) rightward shift of the pressure-flow relation in hypertrophied hearts. which correlated with a greater increase in left ventricular systolic pressure (83 +/- 16 vs 18 +/- 4 mmHg. p < or = 0.05). Exercise resulted in a rightward shift in both normal and hypertrophied left ventricles, but the increase in Pzf was significantly greater in the hypertrophied hearts (15.2 +/- 0.9 vs 10.3 +/- 0.9 mmHg. p < or = 0.05). Stepwise multivariate regression analysis indicated that not only increases in left ventricular filling pressure, but also increases in heart rate and LV systolic pressure contributed to the abnormally great increase in effective coronary back pressure which results in limitation of myocardial perfusion during exercise in the pressure overloaded hypertrophied left ventricle. PMID- 9342436 TI - Ploidy level determination and quantitative chromatin pattern description in pregnancy-associated breast cancers. AB - The present study deals with the characterization of hormone-sensitivity in pregnancy-associated breast cancers (PBCs). This characterization was carried out in 22 PBCs as opposed to 88 non-pregnancy-associated breast cancers (NPBCs). For this study, we used the digital cell image analysis of Feulgen-stained nuclei to assess the type of hormone-sensitivity. In a previous study it was demonstrated that the chromatin pattern in breast cancers is related to the amounts of estrogen receptors they contain. Our results demonstrated that the quantitative description of the chromatin pattern by means of 15 parameters (relating to morphometric, densitometric, and textural features) made it possible to identify typical cell nuclei populations in the PBC and NPBC groups. The use of specific statistical analyses (principal-components and discriminant) made it possible to quantify the proportion of each cell nucleus type in the PBCs. Furthermore, of the 22 PBCs under study, 13 contained a large majority of cell nuclei whose chromatin pattern was characteristic of hormone-sensitive cells, while 5 cases contained a large majority of typically hormone-insensitive ones. The remaining 4 cases contained a relatively similar proportion of typically hormone-sensitive and insensitive cell nuclei. The quantitative chromatin pattern description thus made it possible to characterize the hormone-sensitivity level in PBCs, whereas DNA ploidy level determination did not enable any such characterization to be carried out. The chromatin pattern assay described here, which enables hormone sensitive pregnancy-associated breast cancers to be identified from hormone insensitive ones independently from biochemical assays, should help the physician regarding therapy adaptation. PMID- 9342437 TI - Estrogen sensitivity of normal human breast tissue in vivo and implanted into athymic nude mice: analysis of the relationship between estrogen-induced proliferation and progesterone receptor expression. AB - High serum concentrations of estradiol (E2) equivalent to those observed in the luteal phase of the menstrual cycle stimulate both epithelial cell proliferation and progesterone receptor (PgR) expression in normal human breast tissue xenografted into athymic nude mice. We report here the results of further investigations designed to determine whether the induction of PgR expression and proliferation require different E2 concentrations and whether proliferating cells expressed the PgR. In untreated normal breast xenografts, the PgR was virtually undetectable and proliferation was at basal levels. Progesterone (Pg) treatment alone had no effect compared to no treatment. Treatment with E2 at follicular phase serum concentrations maximally increased PgR expression but was without effect on proliferation. However, treatment with E2 at luteal phase serum concentrations, alone or in combination with Pg, significantly increased both the PgR content and the proliferation of the breast epithelium. These experimentally derived data reflected the observations made on normal breast tissue at surgical biopsy where PgR content was similar in both halves of the menstrual cycle, whereas proliferation was significantly higher in the luteal phase. Finally, using double labelling techniques, it was demonstrated that proliferating epithelial cells rarely expressed PgR in normal breast tissue obtained at surgical biopsy. These results suggest that the threshold of E2 required to induce PgR expression in normal human breast epithelial cells is lower than that required to induce proliferation and that the majority of proliferating breast cells do not express the PgR. PMID- 9342438 TI - Specimen mammography-guided fine-needle aspirates of mammographically normal fatty and dense benign tissue: analysis of cell number and type. AB - Specimen mammography-guided 20-gauge fine-needle aspirates (FNA) were obtained from normal dense or fatty tissue. Single aspirates of dense tissue yielded greater cell counts; 74% (32 of 43) had greater than 5,000 cells as compared to 20% (19 of 93) of fatty tissue FNA, p < 0.05. Dense tissue FNA also yielded greater percentages of epithelial cells; 95% (21 of 22) had greater than 50% epithelial cells as compared to 43% (17 of 40) of fatty tissue FNA, p < 0.05. Mammographic guidance toward dense tissue is suggested for clinical studies of risk assessment using FNA of normal breast tissue. PMID- 9342439 TI - The insulin receptor content is increased in breast cancers initiated by three different oncogenes in transgenic mice. AB - The Insulin Receptor (IR) is a potential oncogene for mammary epithelial cells since its content is increased in most human breast cancer specimens, and both ligand-dependent malignant transformation and ligand-dependent enhanced growth occurs in cultured breast cells overexpressing the IR. To better understand whether the IR plays a role in mammary carcinogenesis which is independent of other initiation factors, we measured IR content in transgenic mouse models of breast cancer induced by 3 known oncogenes (Wnt-1, Neu, and Ret). Insulin receptor content was measured by a specific radioimmunoassay. In normal mammary gland tissues IR content was 14.6 +/- 1.4 ng/mg of protein (mean +/- SEM, n = 6). In the 3 cancers IR content was elevated (Neu = 36.1 +/- 4.6, n = 8, p < 0.002; Wnt-1 = 38.3 +/- 2.6, n = 13, p < 0.001; and Ret = 53.6 +/- 7.1, n = 7, p < 0.001). These data indicate that IR overexpression, in addition to being a potential oncogene, is increased in mouse tumors initiated by other oncogenes, and therefore may also play a supportive role in the growth of breast cancers. PMID- 9342440 TI - Influence of diet on mammary cancer in transgenic mice bearing an oncogene expressed in mammary tissue. AB - Breast cancer is one of the most common cancers in women. The laboratory rat treated with strong carcinogen is the most commonly used animal model for study of breast cancer. Transgenic mouse lines with homologues of human breast cancer oncogenes have been developed. The transgenic mouse line TG.NK with c-neu, the human breast cancer oncogene homologue of erbB2, was evaluated to determine its suitability for study of intervention strategies to delay/prevent the development of breast cancer. There were no palpable mammary tumor masses up to 22-weeks of age, and almost all mice fed a purified diet developed palpable mammary tumors by 28-weeks of age. Nonpurified diets decreased the incidence and multiplicity, and delayed the development of mammary tumors as compared to a purified diet. Increasing the fiber content of nonpurified diet decreased the tumor incidence further. There is approximately a 19-week interval between weaning and development of palpable mammary masses to evaluate intervention strategies to delay or prevent the development of mammary cancer in the TG.NK mouse model. Fiber from nonpurified cereal ingredients appears to be highly beneficial in delaying the development of mammary cancer in TG.NK mice, and this observation is in agreement with human epidemiological findings. Therefore, the TG.NK transgenic mouse with oncogene c-neu (erbB2), appears to be a useful animal model for evaluation of dietary intervention strategies. PMID- 9342441 TI - Increased surgery-induced metastasis and suppressed natural killer cell activity during proestrus/estrus in rats. AB - We have previously reported sex- and estrous-related differences in host resistance to the metastatic development of a mammary adenocarcinoma cell line, MADB106, in the Fischer 344 (F344) rat. In other studies, we found that surgery suppressed natural killer (NK) cell activity and increased the NK-sensitive metastatic development of MADB106 tumor cells. The current study was designed to explore whether sex or estrous phase at the time of surgery impacts the degree of such deleterious effects of surgery. Such estrous effects could be related to an ongoing clinical debate regarding the importance of the timing of breast cancer surgery with the menstrual cycle in premenopausal women. Mature F344 males and cycling females underwent either experimental laparotomy with halothane anesthesia, halothane anesthesia alone, or were untreated. Five hours after surgery, animals either were injected with radiolabeled MADB106 tumor cells and assessed for lung tumor cell retention 12 hours later, or underwent blood withdrawal for in vitro assessment of NK cell activity. MADB106 tumor cells metastasize only to the lungs, and lung tumor cell retention is: a) an early indicator of the number of metastases that would develop weeks later, and b) highly sensitive to in vivo levels of NK activity. This mammary adenocarcinoma cell line is syngeneic to the inbred F344 strain of rats used in our studies, thus constituting a model for breast cancer metastasis. The results indicated that sex, estrous phase, and surgery interacted in their effects on NK cell activity and tumor metastasis. MADB106 lung tumor cell retention was increased by surgery in both sexes (2- to 3-fold) compared to the anesthesia only and control groups. This increase, however, was significantly greater in proestrus/estrus (P/E) females than in metestrus/diestrus (M/D) females. Among the control animals, females in P/E exhibited significantly less NK cytotoxic activity compared to the males, and the NK activity exhibited by females in M/D was between these two groups. Surgery suppressed NK cytotoxic activity to a similar level in all groups. Possible implications of these findings for the surgical care of women with breast cancer are discussed. PMID- 9342442 TI - Similarity between natural and recombinant human alpha-fetoprotein as inhibitors of estrogen-dependent breast cancer growth. AB - Alpha-fetoprotein (AFP) isolated from rodent amniotic fluid or human cord sera, upon incubation with a molar excess of estradiol, is converted to a form which inhibits estrogen-stimulated tissue growth. The purpose of this study was to determine whether recombinant human AFP produced in an E. coli expression system retained this function. The recombinant protein was similar to the natural protein isolated from pooled human cord sera in all functional aspects evaluated. It was detected by monoclonal and polyclonal antibodies to the natural protein. Following exposure to estradiol, it was converted to an inhibitor of estrogen stimulated growth of immature mouse uterus yielding a dose/response curve similar to that of the natural protein. It inhibited the growth of estrogen-dependent (MCF-7) but not estrogen-independent (MDA-MB-231) breast cancer xenografts with the same schedule dependency and resultant histological changes as the natural protein. Availability of large quantities of homogeneous, biologically active recombinant human AFP will facilitate further studies of structure/function, mechanism, and therapeutic potential of this agent as a regular of breast cancer growth. PMID- 9342443 TI - Pattern of dissemination and survival following isolated locoregional recurrence of breast cancer. A prospective study with more than 10 years of follow up. AB - PURPOSES: The study evaluated prognostic factors for dissemination and survival in patients with local or regional recurrence of breast cancer. Furthermore, the aim was to define subgroups of patients at different risk of developing metastases in specific anatomical sites. PATIENTS AND METHODS: The study included 140 patients with isolated local or regional node recurrence, who entered a prospective study for staging of patients with first recurrence of breast cancer in the period 1983-85. The primary treatment was a simple mastectomy; node positive patients received adjuvant radiotherapy and chemotherapy or tamoxifen. If possible, the locoregional recurrence was treated with surgery and/or radiotherapy, otherwise by systemic therapy. RESULTS: Median follow up was 10.4 years; 78 patients developed distant metastases (soft tissue, 32%; bone, 45%; viscera 40%). Median time to dissemination was 4.4 years, and the ten year dissemination rate was 72%. Median time to dissemination was 3.7 years for patients with recurrence in the regional nodes compared to 6.5 years for patients with chest wall recurrence only, p = 0.05. No specific time sequence (temporal pattern) was observed in the anatomical distribution of metastases, and the anatomical site of recurrence could not be predicted by any of the prognostic factors. At follow up, 93 patients had died. The median survival was 5.6 years and 30% were alive after 10 years. Forty-three of the 99 patients who received local therapy only did not develop metastases. Fifteen of these patients died without evidence of metastatic disease while 28 patients were still alive without distant recurrence after a median follow up time of 9.3 years (range, 6.5-11.9 years). Level of LDH and the number of positive regional nodes (NPOS) at primary diagnosis were significant independent prognostic factors for survival after recurrence. CONCLUSIONS: Approximately one third of the patients receiving local treatment only, were alive and without distant metastases up to ten years after locoregional recurrence, indicating that there is a subset of patients which may be long term survivors after local treatment only (surgery or radiotherapy). The duration of survival can be estimated by LDH and NPOS, but the model needs validation in a separate data set before clinical use. PMID- 9342444 TI - Computer simulation of a breast cancer metastasis model. AB - Recent analysis of relapse data from 1173 untreated early stage breast cancer patients with 16-20 year follow-up shows that frequency of relapse has a double peaked distribution. There is a sharp peak at 18 months, a nadir at 50 months and a broad peak at 60 months. Patients with larger tumors more frequently relapse in the first peak while those with smaller tumors relapse equally in both peaks. No existing theory of tumor growth predicts this effect. To help understand this phenomenon, a model of metastatic growth has been proposed consisting of three distinct phases: a single cell, an avascular growth, and a vascularized lesion. Computer simulation of this model shows that the second relapse peak can be explained by a steady stochastic progression from one phase to the next phase. However, to account for the first relapse peak, a sudden perturbation of the development at the time of surgery is necessary. Model simulations predict that patients who relapse in the second peak would have micrometastases in states of relatively low chemosensitivity when adjuvant therapy is normally administered. The simulation predicts that 15% of T1, 39% of T2, and 51% of T3 staged patients benefit from adjuvant chemotherapy, partially offsetting the advantage of early detection. This suggests that early detection and adjuvant chemotherapy may not be symbiotic strategies. New therapies are needed to benefit patients who would relapse in the second peak. PMID- 9342445 TI - Evaluation of the arrhythmogenicity of a low dose of acepromazine: comparison with xylazine. AB - Alteration in the arrhythmogenic dose of epinephrine (ADE) was determined in 6 healthy dogs under halothane anesthesia following the administration of xylazine at 1.1 mg/kg i.v. and acepromazine at 0.025 mg/kg i.v. The order of treatment was randomly assigned with each dog receiving both treatments and testing was carried out on 2 separate occasions with at least a 1 wk interval. The ADE determinations were made prior to drug administration during halothane anesthesia (CNTL) and then 20 min and 4 h following drug treatment. Epinephrine was infused for 3 min at increasing dose rates (2.5, 5.0, 10.0 micrograms/kg/min) until the arrhythmia criterion (4 or more intermittent or continuous premature ventricular contractions) was reached within the 3 min of infusion or the 1 min following cessation. The interinfusion interval was 20 min. There was a significant difference (P = 0.0001) in the ADE determined following acepromazine administration at 20 min (20.95 micrograms/kg +/- 2.28 SEM) compared to CNTL (6.64 micrograms/kg +/- 1.09), xylazine at 20 min (5.82 micrograms/kg +/- 0.95) and 4 h (6.13 micrograms/kg +/- 1.05), and acepromazine at 4 h (7.32 micrograms/kg +/- 0.34). No other significant differences existed (P < 0.05). In this study we were unable to show any sensitization to epinephrine following xylazine administration during halothane anesthesia, while a protective effect was shown with a low dose of acepromazine. PMID- 9342446 TI - Determination of a sedative dose and influence of droperidol and midazolam on cardiovascular function in pigs. AB - Twelve pigs were randomly assigned to 1 of 2 groups, droperidol or midazolam, to determine a sedative dose of each drug that would facilitate handling of the pigs. Each pig in the group received all of the test doses with 5-7 d between treatments (droperidol--0.1, 0.3, 0.6 mg/kg, or midazolam--0.25, 0.5, 1.0 mg/kg) and saline (3 mL), i.m. One investigator, unaware of the treatment administered, assessed the time of onset, degree, and duration of sedation. The 0.3 mg/kg dose of droperidol and 0.5 mg/kg dose of midazolam were judged to be the most suitable for sedation and produced similar degrees of sedation, although the onset and duration of sedation was significantly longer for the droperidol group. The effects of these 2 doses on heart rate, respiratory rate, systolic blood pressure, and rectal temperature were assessed in 12 pigs randomly assigned to 1 of the 2 treatments. Respiratory rate decreased significantly with droperidol at 10, 15, and 30 min. Temperature was significantly decreased at 60 min following midazolam. This study demonstrates that 0.3 mg/kg i.m. of droperidol and 0.5 mg/kg i.m. of midazolam induce adequate sedation in pigs with minimal cardiorespiratory changes. PMID- 9342447 TI - Pharmacokinetics of oral morphine sulfate in dogs: a comparison of sustained release and conventional formulations. AB - The pharmacokinetics and bioavailability (F) of single dose sustained release morphine sulfate (OSRMS) and nonsustained release morphine sulfate (NSRMS) were compared to each other and to a bolus injection of morphine sulfate (MS) intravenously (i.v.) in dogs. Beagles (n = 6) were randomly assigned to 3 treatment groups: namely, OSRMS 15 mg orally, NSRMS 15 mg orally, and 15 mg i.v. Serum samples were drawn at intervals up to 480 min following oral and 420 min following i.v. administration. Serum was analysed for morphine concentration using a radioimmunoassay. Data were analysed using non-compartmental pharmacokinetics. The only statistically significant difference between OSRMS and NSRMS was maximum serum concentration (Cmax). There were trends toward longer time to maximum serum concentration (Tmax) and longer mean absorption time (MAT) for OSRMS when compared to NSRMS, but the differences were not statistically significant (P < 0.05). Pharmacokinetic parameters for both oral formulations exhibited large variability in the rate of absorption of MS from the gastrointestinal tract. Bioavailability of both OSRMS and NSRMS was low (15% 17%). As expected, the area under the concentration vs time curve (AUC) and Cmax for the i.v. data was significantly greater than for both oral groups, and Tmax and mean residence time (MRT) were significantly less following i.v. administration. There were no statistically significant differences among the 3 treatment groups for apparent volume of distribution at steady state (Vdss) or elimination parameters. The OSRMS formulation used in this study provided equivalent bioavailability to NSRMS in dogs, accompanied by large individual variability in drug absorption. It also did not appear that the sustained release formulation provided sufficiently prolonged release of morphine sulfate from the tablet matrix in dogs to allow prolonged dosing intervals compared to NSRMS. PMID- 9342448 TI - Status of Trichinella spiralis in domestic swine and wild boar in Canada. AB - Evidence of the status of trichinellosis in Canada's national swine herd is provided from data acquired through national surveillance programs and from a prevalence study of Trichinella in wild boar and domestic swine. More than 500,000 swine tested at abattoirs in ongoing animal health surveys since 1980 and 2 national swine serological surveys (1985 and 1990) showed no evidence of Trichinella infection, except for 3 occurrences in a small infected zone in Nova Scotia. The prevalence study of domestic swine and wild boar was conducted for the prevalence of Trichinella after an epidemiological investigation of a 1993 outbreak of human trichinellosis in Ontario showed that the disease was linked to the consumption of wild boar meat originating from 2 farms in the province. Sera and tissues were collected from 391 wild boar and 216 domestic swine originating from 228 farms in Quebec, Ontario, Manitoba and Saskatchewan. The survey examined approximately 37% of the wild boar slaughtered in Canada in 1994. A pepsin-HCl digestion test of the tissues and an ELISA performed on the sera did not yield any positive results. These findings and the lack of human cases of Trichinella from the consumption of Canadian pork for nearly 2 decades suggest that the parasite has been rare in domestic swine and wild boar raised in Canada. Trichinella spiralis has only been found sporadically in swine in a small region within Nova Scotia. PMID- 9342449 TI - Development and initial evaluation of an indirect enzyme-linked immunosorbent assay for the detection of Leptospira interrogans serovar hardjo antibodies in bovine sera. AB - Outer sheath antigen from Leptospira interrogans serovar hardjo type hardjoprajitno and acetic acid extracted antigens from serovar hardjo types hardjoprajitno and hardjobovis were evaluated in an immunoassay for ability to detect hyperimmune rabbit serum to serovar hardjo. The degree of cross-reactivity with hyperimmune rabbit sera to L. interrogans serovars pomona, copenhageni, grippotyphosa, canicola and sejroe, and Leptospira biflexa serovar patoc was also measured for each antigen. All of the antigens reacted with the antiserum to L. interrogans serovar hardjo. The outer sheath antigen however, also showed wide cross-reactivity with the antisera to all of the serovars of L. interrogans tested and with the antiserum to L. biflexa serovar patoc. The acetic acid extracted antigen from either type hardjoprajitno, or type hardjobovis, showed a high degree of specificity for serovar hardjo antiserum. The hardjobovis acetic acid extracted antigen was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting, and was incorporated into an indirect ELISA for detection of anti-serovar hardjo antibodies in bovine serum. This ELISA showed a relative specificity of 100% with 156 bovine sera which were negative at a dilution of 1:100 in the microscopic agglutination test (MAT) for L. interrogans serovars hardjo, pomona, sejroe, icterohaemorrhagiae, copenhageni, canicola, and grippotyphosa. The relative sensitivity of this assay with 192 bovine sera which had serovar hardjo MAT titres of > or = 100 was 95.3% (95% confidence limit = 2.99%). The degree of cross-reactivity with 289 bovine sera which had serovar pomona MAT titres of > or = 100 (with no detectable serovar hardjo MAT titres) was approximately 1.0%. This assay was: easily standardized, scored objectively, repeatable, semi-automated and used a non-hazardous antigen that can be routinely prepared in gram amounts. PMID- 9342450 TI - Development of a monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the detection of Leptospira borgpetersenii serovar hardjo type hardjobovis antibodies in bovine sera. AB - A murine monoclonal antibody (designated M553) that binds to an epitope on whole cell antigens prepared from Leptospira borgpetersenii serovar hardjo type hardjobovis and Leptospira interrogans serovar hardjo type hardjoprajitno, was produced and incorporated into a competitive enzyme-linked immunosorbent assay for the detection of bovine antibodies to serovar hardjo. The epitope recognized by M553 was susceptible to periodate oxidation. The M553 antibody was characterized by western blot with hardjobovis whole cell antigen. This antibody does not cross-react with whole cell antigens prepared from 11 other pathogenic Leptospira serovars, or, Leptospira biflexa serovar patoc. The sensitivity estimate of the competitive ELISA was 100% with field sera (n = 165) with serovar hardjo microscopic agglutination test (MAT) titres of > or = 100. The specificity estimate was 100% with sera (n = 128) obtained from a specific pathogen free herd of cattle that were negative in the MAT at a dilution of 1:100 for serovars hardjo, pomona, sejroe, copenhageni, canicola, and grippotyphosa. The specificity estimate with field sera (n = 301) with serovar hardjo MAT titres of < 100, was 98% (95% confidence interval = +/- 1.58%). There was no cross-reactivity with field sera (n = 306) with serovar pomona titres > or = 100 and serovar hardjo titres < 100. The specificity estimate with the combined populations of sera with serovar hardjo MAT titres of < 100 (n = 735) was 99.18% (95% confidence interval = +/- 0.65%). There was a high level of agreement (kappa = 0.977) between the results of the competitive ELISA and those of the MAT. PMID- 9342451 TI - Immunization of pigs against Streptococcus suis serotype 2 infection using a live avirulent strain. AB - Streptococcus suis capsular type 2 is still an important cause of economic losses in the swine industry. At the present time, vaccination of pigs against this infection is generally carried out with autogenous bacterins and results are equivocal. In this study, the protective effect of a live avirulent S. suis type 2 strain (#1330) which had induced a good protection in mice, was evaluated in swine. The experiment was performed in triplicate using 4 week-old piglets. A total of 15 piglets were vaccinated 3 times, 15 others were vaccinated 2 times, and 15 piglets were injected 3 times with sterile Todd-Hewitt broth. Using an indirect ELISA, an increase in the IgG response to S. suis antigens was noted in 27 of the 30 vaccinated piglets. On day 21 post-vaccination, all animals were challenged intravenously with a virulent S. suis type 2 strain (#999). In the 2 vaccinated groups, 26 animals were fully protected. Only 1 out of the 15 piglets vaccinated 3 times developed mild clinical signs. In the group vaccinated twice, 3 piglets showed clinical signs and 1 of them died after the challenge. In the control group, 7 animals died out of the 11 with clinical signs of infection. In conclusion, a protective immunity was observed in swine when using strain 1330. However, more studies are needed to assess the use of a live S. suis strain in a vaccine for pigs. PMID- 9342452 TI - Prevention of edema disease in pigs by vaccination with verotoxin 2e toxoid. AB - Pigs in 2 herds with persistent problems with post weaning edema disease caused by infection with verotoxin-2e (VT2e)-producing Escherichia coli O139 were treated with a VT2e-toxoid vaccine. Treatment was performed as a randomized blind field trial with parallel treatment and non-vaccinated control groups. In 1 herd, a group of pigs was injected with adjuvant alone. Pigs were vaccinated at 1 and 3 wk of age and weaned at 4 wk of age. The effect of vaccination was measured by average daily weight gain (ADG), mortality due to edema disease within the 1st 4 wk after weaning, and weight at 3-6 mo of age. Pathological and microbiological examinations were performed on all pigs that died during the 1st 4 wk post weaning. Only pigs from which VT2e+, F18+ E. coli O139 was isolated were categorized as "death due to edema disease." The serological response to vaccination was evaluated by an indirect ELISA. Vaccination had a statistically significant effect on the level of antibodies specific for VT2e in both herds. Vaccination resulted in a statistically significant increase in ADG in the nursery period but not in the grower-finishing period. Vaccination had a statistically significant effect on mortality due to edema disease with an odds ratio of 0.039, indicating that there was almost total elimination of mortality due to the disease in the vaccine groups. PMID- 9342453 TI - Production of a baculovirus-derived gp50 protein and utilization in a competitive enzyme-linked immunosorbent assay for the serodiagnosis of pseudorabies virus. AB - The pseudorabies virus (PRV) gp50 envelope glycoprotein gene was cloned and expressed in a recombinant baculovirus. An anti-gp50 Mab (1842) recognized a protein of approximately 40 kDa in immunoblotting assays from infected insect cell lysates, while this product was not present in cells infected with wild-type baculovirus. The recombinant protein was purified by lectin affinity chromatography, utilizing lectins specific for O-linked oligosaccharides (Artocarpus integrifolia and Glycine max). Competitive (c) ELISAs, using either crude or lectin-purified antigen, were devised for the detection of antibodies to PRV in sera, and were capable of monitoring sero-conversion by day 14 post infection. Furthermore, a specificity of 100% and sensitivity of 98% (crude lysate antigen) or 96% (lectin-purified antigen) was found for a panel of 80 swine sera, using the cELISA, as compared to a serum neutralization (SN) test. These studies demonstrated that recombinant PRV gp50 protein shows promise as a cELISA antigen, for serodetection of PRV. PMID- 9342454 TI - Persistence of porcine reproductive and respiratory syndrome virus in intensive farrow-to-finish pig herds. AB - An epidemiological study of porcine reproductive and respiratory syndrome (PRRS) within pig herds was conducted in 8 intensive farrow-to-finish pig farms. Persistence of PRRS virus (PRRSV) in pig herds was demonstrated by regular postmortem examination on 2 farms for a period of 2 y. Virus isolation and serum neutralization (SN) tests were performed on the sera collected from 9 groups of pigs (10 pigs/group) of various ages on 8 pig farms. Except for 1 farm, isolation rates of PRRSV reached the highest level of 70 to 100% of pigs 6 to 8 wk of age, which coincided with the lowest levels of maternal immunity. In 1 pig herd, sows (39 in total) with SN titers of < or = 1:2, 1:4-1:8, and > or = 1:16 were designated as groups 1, 2, and 3, respectively. Sera were obtained from their progeny (3 pigs randomly selected from each litter) at various ages from 0 to 22 weeks. A positive correlation (r = 0.377, P < 0.001) between the SN titers of sows and those of their progeny (1-week-old piglets) was observed. Pigs at the age of 6 wk, only 7.9% of group 1 pigs compared to 72.4% of group 3 pigs were seropositive. A significant difference (P < 0.01) in the percentage of pigs with PRRSV viremia among the 3 groups was observed, with the lowest level found in group 3 pigs. The isolation rates of PRRSV from serum reached the maximum at the age of 9 wk for all 3 groups. The results indicated that passively acquired serum antibodies conferred a protective effect for piglets; however, loss of passive immunity at various ages of pigs produced susceptible pigs that resulted in PRRSV persistence in the pig herds. Pigs 6 to 9 weeks old were the major reservoir for PRRSV in farrow-to-finish pig herds. PMID- 9342455 TI - Performance of ELISA antigens prepared from 8 isolates of porcine reproductive and respiratory syndrome virus with homologous and heterologous antisera. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) ELISA antigens of high quality were produced using 8 different isolates of PRRSV: the European Lelystad virus (LV), the U.S. MN-1b, 89-46448, 93-44927, and 93-24025B, and the Canadian LHVA-93-3, PA-8 and GH-6 virus isolates. The performance of each of these 8 antigens and a commercial PRRSV antibody test kit (Idexx's HerdChek) were measured against antisera raised in 5 groups of 6 piglets inoculated with either LV, MN-1b, 89-46448, 93-44927, or 93-24025B. Among the 8 isolates, the 89-46448 isolate produced the broadest spectrum of antigen and resulted in earlier detection of antibodies to various North American PRRSV isolates, followed by MN 1b as the 2nd best ELISA antigen for the detection of North American PRRSV antibodies. The GH-6 and PA-8 viral antigens exhibited restricted detection of PRRSV antibodies. The LV and 89-46448 combined antigens produced the best performance for the detection of antibodies against both European and North American antigenic types of PRRSV. Using 173 panel samples collected at 11 to 60 d after intranasal inoculation with 1 of the 5 PRRSV isolates, the sensitivities of the indirect ELISA used were 73.4%, 98.3%, 90.8%, 98.3%, 83.2%, 93.1%, 77.1%, 64.2%, 98.8% and 95.9% for LV, MN-1b, LHVA-93-3, 89-46448, 93-44927, 93-24025B, PA-8, GH-6 antigens, 89-46448-LV combined antigens and Idexx's PRRSV antibody test kit, respectively. All 8 antigens gave negative results with preinfection porcine sera (n = 30); high background or nonspecific reactions were not observed with the antigens. PMID- 9342456 TI - Prevalence of coronavirus antibodies in Iowa swine. AB - Three hundred and forty-seven serum samples from 22 Iowa swine herds were screened for TGEV/PRCV neutralizing antibody. Ninety-one percent of the sera and all 22 herds were positive. These sera were then tested by the blocking ELISA test to distinguish TGEV and PRCV antibody. The ELISA test confirmed the high percentage of TGEV/PRCV positive sera. By the blocking ELISA test, 12 herds were PRCV positive, 6 herds were TGEV positive and 4 herds were mixed with sera either positive for TGEV or PRCV antibody. The results suggest a recent increase in TGEV/PRCV seroprevalence in Iowa swine most likely due to subclinical PRCV infections. PMID- 9342457 TI - Rate of decline of cortisol concentrations in ovarian follicles following ACTH treatment in the sow. AB - The rates of decline in cortisol concentrations in blood and ovarian follicular fluid were assessed in cyclic sows (n = 30) after treatment with saline or a depot form of adrenocorticotrophic hormone (ACTH). After a single injection of ACTH (0.5 iu/kg, BW, i.m.), peak cortisol values were achieved in blood within 3 to 4 h followed by a half-life net clearance rate (t1/2 of 2.40 +/- 0.29 (SE) h. The same dose of ACTH was then given at 12 h intervals from days 9 to 13 of the estrous cycle. On day 14 the concentrations of cortisol in follicular fluid were higher (P < 0.05) in ACTH-injected sows than in saline-injected controls. A t1/2 value of 37.81 h was determined for cortisol based on the decline in concentrations in follicular fluid collected on days 14, 16 and 18. This relatively slow rate of removal from developing ovarian follicles may have implications for the previously observed detrimental effects of increased cortisol concentrations on follicular development. PMID- 9342458 TI - Reduced concentrations of apolipoproteins B-100 and A-I in serum from cows with retained placenta. AB - The purpose of the present study was to evaluate apolipoprotein B-100 and A-I concentrations in cows with retained placenta. Animals used were cows with retained placenta alone (n = 10), those with both retained placenta and ketosis (n = 7), and controls (n = 10). Apolipoprotein B-100 concentrations at 2 to 4 d after parturition were significantly (P < 0.01) decreased in cows with retained placenta alone (mean +/- SD, 0.084 +/- 0.029 mg/ML of serum) when compared with those in control cows (0.154 +/- 0.022 mg/mL). Apolipoprotein A-I concentrations (0.713 +/- 0.177 mg/ML) were also significantly (P < 0.05) lower than those of controls (0.895 +/- 0.159 mg/mL). These decreases were more distinct for apolipoproteins B-100 (55% of controls) than A-I concentrations (80% of controls). Concentrations of apolipoprotein B-100 (0.071 +/- 0.032 mg/mL; P < 0.01) and A-I (0.708 +/- 0.189 mg/mL; P < 0.05) in the cows with both retained placenta and ketosis were also reduced, when compared with values in controls. Other than apolipoproteins, cows with retained placenta alone had significantly (P < 0.01) higher serum nonesterified fatty acids, and lower triglyceride concentrations. Significantly (P < 0.01) higher nonesterified fatty acids and lower triglyceride concentrations were similarly observed in cows with both retained placenta and ketosis. PMID- 9342460 TI - An eight year review of hospitalization for ovarian hyperstimulation syndrome. AB - We analyzed the etiologic factors and trends of hospitalization for ovarian hospitalization syndrome (OHSS) resulting from the use of fertility medications. From May, 1986 through April, 1994, patients hospitalized with OHSS were exclusively admitted to the University Hospital. Analysis was performed with regards to treatment method, severity of hyperstimulation, and onset of disease. Overall, 14 patients were hospitalized for a rate per cycle of 0.1% (14/14, 283). The rate of admission for patients undergoing superovulation (9/12, 945; .07%) was significantly lower than for those undergoing assisted reproductive techniques (ART) (5/1, 338; .37%). The total number of injectable gonadotropin ampules used was also higher in patients admitted following ART versus superovulation. A significantly greater number of patients presenting with late developing hyperstimulation syndrome (5/7; 71.4%) manifested severe disease as opposed to those hospitalized with early onset OHSS (1/7; 14.3%). Our data suggest that hospital admission is an infrequent event following the use of fertility medications, and patients are more likely to be hospitalized with OHSS following ART than superovulation. PMID- 9342459 TI - Dose related mucosal hyperplasia induced by Oesophagostomum dentatum infection in pigs. AB - The present work was undertaken to examine the effects of 3 different population densities of Oesophagostomum dentatum upon the development of worm induced mucosal changes in the colon following single infections. Groups of pigs were infected with single doses of 2000 (low dose), 20,000 (medium dose) or 200,000 (high dose) infective larvae, respectively. A total of 18 infected pigs (6 from each group) were examined for histopathological changes together with 3 helminth free control pigs. There was a dose related difference in the intensity of colonic lesions; and using morphometry it was observed that the mucosal crypts of pigs in the high dose group were significantly longer than those in the 2 other groups. These differences disappeared by day 25 after infection despite the presence of larvae in the mucosa of the high dose group. This phenomenon may be related to inflammatory reactions in the colon, possibly in connection with the initiation of an immunological response in sites distant from the parasite larvae. PMID- 9342461 TI - Effects of maternally administered immunoglobulin on platelet counts of neonates born to mothers with autoimmune thrombocytopenia: re-evaluation. AB - OBJECTIVE: Since immunoglobulin is transported across the placenta, maternal administration of it theoretically seems attractive as an antenatal treatment for a fetus. However, the effects of antenatally administered intravenous immunoglobulin (IVIG) on fetal platelet counts have been controversial. Our series of 11 cases of idiopathic thrombocytopenic purpura (ITP) and a review of previous reports are presented. A combined retrospective analysis to know whether maternal response to IVIG is associated with improvement of fetal thrombocytopenia was conducted. METHODS: IVIG was given to 11 steroid refractory pregnancies with ITP. Good maternal response to the therapy was defined as an increase in the platelet count to greater than 50 x 10(9)/L after completion of the IVIG infusion. Neonatal platelet counts of the umbilical cord were performed just after birth and followed-up for at least the first week of life. RESULTS: Seven of the 11 neonates had thrombocytopenia of less than 100 x 10(9)/L, and two of them had severe thrombocytopenia of less than 50 x 10(9)/L. Two out of two neonates born to mothers with a good response to IVIG were thrombocytopenic; whereas five out of nine neonates born to mothers with a poor response were thrombocytopenic. The combined retrospective analysis of our results and published reports have shown that fetal thrombocytopenia was not associated with the maternal response to IVIG but with the level of immunoglobulin G of neonates at birth. Passive thrombocytopenia was more frequently observed in neonates with normal levels of immunoglobulin G than those with elevated levels. CONCLUSION: This combined analysis confirmed the proposed hypothesis that the lack of effect of maternally administered IVIG on fetal platelet counts may mainly be attributed to the insufficient therapeutic level of transferred immunoglobulin G in the cord blood. PMID- 9342462 TI - Prevention of thromboembolic complications in women undergoing gynecologic surgery. AB - Our objective was to identify those patients particularly at risk of deep vein thrombosis (DVT) before they underwent extensive gynecologic surgery and to control if, a correct diagnostic analysis and a right pre-operative prophylaxis of patients with risk of developing DVT, was enough to improve post-operative prognosis. Of 2704 patients undergoing gynaecological surgery, 74 were pre operatively considered at risk of developing DVT. Seventy percent of the patients received pre- and postoperative heparin, while 28% of the women received only postoperative heparin. Nonetheless, seven women receiving this prophylaxis developed DVT. The final results of our study demonstrate that there is a close correlation between incidence of DVT and the presence of risk factors. This incidence can be reduced by prophylactic measures such as elastic stockings for the lower legs, early post-operatory mobilization, hematocrit and volemy control, ending with pharmacological therapy with heparin. PMID- 9342463 TI - Eligibility criteria for labor induction with prostaglandins. AB - Particular conditions exist at the end of some pregnancies which cause an increase in maternal and fetal risk. A valid alternative for these pregnancies is represented by the administration of prostaglandins, in order to obtain labor induction. The goal of our study was to define the eligibility criteria and the epidemiological characteristics that correlate most with a favorable obstetrical outcome. The study was conducted on 133 informed, consenting patients subjected to labor-induced delivery with prostaglandins E2. The mode of delivery in relationship to parity demonstrated that the pluriparous patients had fewer difficulties in labor and in its induction: of the 43 pluriparous cases, none had a cesarean section for failed induction and 95.3% delivered vaginally. One hundred percent of the patients with a Bishop score of more than 4 went into labor, as opposed to 81% of the patients with a score of less than 4. Therefore, taking into consideration the cost of the method, we retain that choosing an active position is valid, respecting the eligibility criteria for the induction of labor described above. PMID- 9342464 TI - The effect of antigamete antibodies on the success of assisted reproduction. AB - The aim of the study was to investigate the presence of antigamente antibodies in unexplained infertility patients and to prove the efficiency of IUI and IVF-ET treatments for these patients. The study includes 46 unexplained infertility patients and as controls, a group of 21 tubal infertility patients. Serum, follicular fluid and cervical mucus samples were collected from each patient and antibodies were measured with commercial ELISA kits. Twenty-two of the 46 unexplained infertility patients produced at least one of the antibodies against sperm or ovary. Fertilization rates were lower in immunological and unexplained infertility patients than in tubal infertility patients, being statistically significant. Pregnancy rates were lower in immunological and unexplained infertility patients than in tubal infertility patients after IVF-ET, but this was not statistically significant. Pregnancy rates after IUI treatment were equal in both immunological and unexplained infertility groups. AGA (antigamete antibodies) were found in 45% of unexplained infertility patients and therefore may be a possible cause of infertility. IUI and IVF-ET are successful choices for treatment of these patients. PMID- 9342465 TI - Amniotic fluid index variations after amniocentesis, amnioinfusion and amnioreduction: preliminary data. AB - We studied the relationship between the ultrasonographically measurable variations in the amniotic fluid index (AFI) and actual changes in the amniotic fluid volume induced by three differing invasive procedures: genetic amniocentesis, amnioinfusion and amnioreduction. We examined 50 patients, all between the 15th and 34th weeks of pregnancy, subdivided into three groups. The first group consisted of 33 women who underwent genetic amniocentesis, the second was of 11 patients submitted to amnioinfusion for oligohydramnios (AFI < 5 cm), and the third was composed of 6 patients affected by hydramnios (AFI > 20 cm) and treated with amnioreduction. In all cases AFI was measured before and after the invasive procedures and their variations (delta AFI) were correlated to the actual quantities of liquid infused or extracted. All the procedures gave rise to statistically significant AFI changes. After genetic amniocentesis, the mean change was from 12.0 to 10.9 cm (p < 0.005), after amnioinfusion from 3.1 to 10.6 cm (p < 0.0001) and after amnioreduction from 33.1 to 22.0 cm. (p < 0.005). However, a significant linear correlation between delta AFI and the fluid volume variations actually induced was found for amnioinfusion (y = 0.236537 + 0.031465x; R2 = 44.4%; p < 0.05) and for amnioreduction (y = -0.0584294 + 0.012008x; R2 = 89.8%. p < 0.00001). Only for amnioreduction is it possible, as proved by a multiple regression analysis, to improve the predictability of delta AFI, taking into consideration together with the quantity of fluid aspirated, the value of the preprocedure AFI (R2 = 92%; p < 0.05). PMID- 9342466 TI - Hysterectomy in women with cervical pregnancy complicated by life-threatening bleeding: a case report. AB - A 27-year-old women, gravida 2, para 1 presented with massive vaginal bleeding. After two days of bleeding from the external cervical ostium, intracervical tamponade was performed but the bleeding did not stop. At laparotomy, abdominal hysterectomy with adnexa preservation was done because of malacia tissue and life threatening hemorrhage. Pathological examination revealed an isthmic pregnancy, gravidic decidua, and chorion villi. PMID- 9342467 TI - Surgical management of leiomyomata in pregnancy. AB - A review was made of the medical records of 26 patients with uterine myomas during pregnancy between 1983 and 1992 among 12,965 deliveries. Thirteen patients underwent myomectomies before pregnancy. In three patients myomectomy was performed during pregnancy between the 12th and the 19th week of pregnancy. In ten patients myomectomy was performed during cesarean section delivery to prevent necrobiosis. Myomectomy should remain exceptional during pregnancy and it must be performed only in selected cases but is frequently used towards the end of a cesarean section. Indications for hysterectomy, on the other hand, remain limited. PMID- 9342468 TI - Non-stress test: a fifteen-year clinical appraisal. AB - During a 15-year period (Jan. 1981 to Dec. 1995) a total of 14,950 patients were delivered in our hospital. Throughout this period fetal heart rate monitoring during labor was increased from 10% up to 85%. The overall antepartum testing was also increased from 8 to 15%. In patients with significant complications in pregnancy the mean number of non-stress tests (NST's) per patient was 1.8 tests in 1981 to 4.8 tests in 1995. The average perinatal mortality was 15.2/1000, with a gradual decline. In patients who were subjected to antepartum testing the previous rate was only 3.7%/1000. Our conclusion is that the use of antepartum and intrapartum cardiotocography has increased during the last 15 years in our clinic. As a consequence a considerable decrease was noted in the overall perinatal mortality. The non-stress test is still today the first line of antepartum fetal assessment. PMID- 9342469 TI - Blood pressure peaks correlated with plasma fibronectin levels and microalbuminuria in hypertensive pregnancies. AB - A group of 32 selected hypertensive pregnant women under antihypertensive therapy, with biochemical parameters, functional parameters, plasma fibronectin levels (PLF), microalbuminuria (MA) levels and continuous 24 h blood pressure monitoring, were followed monthly until delivery and during puerperium. Also possible biochemical and clinical markers and the predictive value in the complications during PIH were attempted to be identified. There was a statistical correlation between systolic pressure peaks associated with high levels of PLF and MA in hypertensive pregnant women who may have a higher risk of pregnancy or cardiovascular complications. Continuous 24 h blood pressure monitoring in hypertensive pregnancies was found to be helpful in identifying the highest risk patients especially by reading the night peak percentages. PMID- 9342470 TI - Penetration of pesticides in the human reproductive system. Presentation of case reports. PMID- 9342471 TI - Does postprandial hypersinsulinemia contribute to hyperandrogenism in patients with polycystic ovary syndrome? AB - Twenty patients with polycystic ovarian syndrome and with or without insulin resistance, and 20 healthy women (controls) underwent an oral glucose tolerance test, which resulted in a short duration but significant increase of serum insulin levels. Serum testosterone, androstenedione and dehydroepiandrosterone sulfate levels were estimated before and 180 minutes after administration of 75 gr. dextrose. Our results, three hours after dextrose administration, showed that: (1) serum testosterone levels decreased significantly, (2) serum androstenedione levels decreased but not significantly, and (3) serum dehydroepiandrosterone sulfate levels were not altered. The observation of decreased ovarian androgen levels after induced hyperinsulinemia is very interesting, the explanation, however, is quite difficult. This unexpected ovarian androgen response needs further investigation. PMID- 9342472 TI - Minimally invasive treatment of live ectopic pregnancy. AB - Four cases of live ectopic (three tubal, one cornual) pregnancies managed by ultrasonographic puncture injection technique are described. The technique proves to be a practical, effective and conservative treatment. PMID- 9342474 TI - Transvaginal Doppler ultrasound with color flow imaging in the diagnosis of luteal phase defect (LPD). AB - OBJECTIVE: Our purpose was to determine whether color flow pulsed Doppler could predict a luteal phase defect (LDP). METHOD: Twenty-one women with regular menstrual cycles and at risk for luteal phase defect were examined by transvaginal color Doppler during the follicular and luteal phase of the menstrual cycle. Progesterone was measured on the day of the Doppler exam. Ovulation was determined from the lutenizing hormone (LH) surge. Endometrial biopsy during the late luteal phase was performed on each patient. RESULT: Six patients (28.5%) were diagnosed with luteal phase defect. Mean resistance index in patients with luteal phase defect was significantly higher only throughout the luteal phases (p = 0.02). Mean progesterone levels were significantly lower for patients with LPD than for normal women (p < 0.001). Mean pulsatility index in luteal phase deficient cycles was significantly higher throughout the follicular and luteal phases (p = 0.03). CONCLUSION: Color Doppler may aid in assessing luteal phase adequacy. Doppler indices of corpus luteum blood flow in combination to plasma progesterone may be a useful index of luteal function. PMID- 9342473 TI - Therapeutic protocol of vulvar and cervical HPV-infection. AB - BACKGROUND: The stages of HPV (Human Papilloma Virus) infection are under the control of the immune system, which is inhibited by the virus itself. Thus, at present the treatment of condyloma acuminata is based on the use of interferon (IFN). The aim of the present study was to evaluate immune system activation and clinical response to IFN therapy. In addition, in the most serious cases, medical treatment with IFN was associated with diathermocoagulation (DTC) of persistent warts. The effectiveness of the combined therapy was also assessed. METHODS: 7 women (age range: 16-52) suffering from cervix condylomata were selected for our study. All of them were injected intramuscularly with doses of 3 million UI of IFN-alpha leucocytar every three days for six weeks together with daily applications of alpha-IFN for six weeks. The women that were still ill three months after IFN therapy, were treated with DTC. In one case, another cycle of IFN treatment was necessary. RESULTS: Clinical response to IFN treatment was complete in 46 cases, partial in 20 cases and unsuccessful in 4 cases. After three months of medical therapy, 30 women were treated with DTC. After this therapy, in 21 cases, the warts were resolved. After 24 months the percentage of relapse was 37.9% when only IFN was used, and 4.51% when IFN was combined with DTC. CONCLUSIONS: These data suggest that a successful protocol for the treatment of condyloma acuminata consist of IFN therapy associated with DTC when warts persisted. PMID- 9342475 TI - Laparoscopic treatment of benign adnexial cysts. AB - The results of laparoscopic adnexial cyst excision operations performed within the last 2.5 years in our clinic are reported. Thirty-three adnexial masses thought to be benign after gynaecological examination and ultrasonographic findings were treated. Laparoscopy was done in 32 cases but laparotomy had to be performed in one case of stage IV endometriosis. The mean duration of the operations was 72.78 +/- 34.09 minutes and no major complication occurred. Pathologic examinations of the specimens were reported as benign in all cases. According to these results, laparoscopy should be the preferred method in the treatment of benign adnexial cysts. PMID- 9342476 TI - Is color Doppler necessary in the evaluation of tubal patency by hysterosalpingo contrast sonography. AB - BACKGROUND: To evaluate the diagnostic efficacy of hysterosalpingo-contrast sonography (HyCoSy) and to establish whether, in doubtful cases, the additional use of transvaginal Color Doppler can improve the tubal diagnosis reached with gray scale imaging alone. STUDY DESIGN: Ninety-six cases of unknown tubal function with infertility complaints were included. Within four weeks after sonographic hydrotubation, hysterosalpingography was performed. The diagnostic efficacy of gray scale and Color Doppler were compared with each other and against HSG. RESULTS: We calculated a sensitivity value of 82.9% and a specificity value of 86.1% for both tubes. The concordance value was of 83.5%. The total number of Color Doppler examinations compared with those of HSG showed a sensitivity of 91% for the right tube and 93% for the left tube and a specificity of 100% for both tubes. CONCLUSIONS: HyCoSy provides reproducible findings for the evaluation of uterine abnormalities and tubal patency. The additional use of Color Doppler is recommended as a supplement to gray scale imagin in cases of suspected tubal occlusion. PMID- 9342477 TI - Cervical cerclage for malformed uterus. AB - The role of cervical cerclage was evaluated in six pregnant women with anomalous uterus. Early prophylactic cerclage according to the Shirodkar and McDonald technique was done on all cases of uterine malformation (except septate uterus) with or without cervical incompetence in association with progesterone and antispastic therapy. Improvement in obstetrical outcome was noted after cerclage. Even if no doubt exists as to the need for cerclage in cases of cervical incompetence, the concept of routine prophylactic cerclage in all cases of uterine anomalies should be considered. PMID- 9342478 TI - Late abdominal pregnancy: expectant management with survival of the infant. PMID- 9342480 TI - Endometrium is not a reservoir for recurrent vaginal candidiasis. AB - The aim of this study was to determine whether the endometrium acts as a reservoir for Candida albicans in cases of recurrent vaginal candidiasis. Twenty five women with documented history of recurrent vaginal candidiasis were enrolled in the study and endometrial samples were cultured for Candida albicans. Only two patients had positive cultures for Candida albicans. Therefore, we concluded that the endometrium is not a common reservoir for Candida albicans. PMID- 9342479 TI - Cardiotocographic and Doppler velocimetric patterns, pre- and post-thoracentesis, in a case of fetal hydrothorax. AB - Fetal hydrothorax is associated with elevated perinatal mortality. Management of this condition is controversial given that in utero spontaneous resolution has been described. A case of fetal hydrothorax associated with an extralobar lung sequestration that showed pathologic cardiotocographic patterns and abnormal Doppler velocimetry indices in several fetal vascular beds in reported. All pathologic patterns improved after fetal thoracentesis. It can be concluded that monitoring fetal well-being by means of cardiotocography and Doppler velocimetry may help in timing thoracentesis in cases of fetal hydrothorax. PMID- 9342481 TI - Pregnancy in a patient with essential thrombocytosis. AB - An insurgent case of pregnancy in a patient in whom essential thrombocytosis was diagnosed five years earlier is described. Pregnancy was confirmed and therapy with platelet aggregation inhibitor was introduced. The pregnancy reached full term notwithstanding a positive result of the "Triple Test" during the 15th week of gestation. A histology exam of the placenta revealed an ischemic lesion. We retain that platelet aggregation inhibitor therapy remains an important aid in eliminating the risk of thrombosis determined by the presence of two conditions that are predisposed to these risks, such as pregnancy and essential thrombocytosis. PMID- 9342482 TI - Prolapse of the fallopian tube into the vaginal vault. PMID- 9342483 TI - Glomerular basement membrane antibodies in hantavirus disease (hemorrhagic fever with renal syndrome). AB - The prevalence of antibodies against glomerular basement membrane (GBM) antigens in sera from 47 patients with serologically verified hantavirus (Puumala serotype) infection were investigated. Antibodies were measured by immunoassays using as antigen a crude human GBM preparation and the NC1 portion of type IV collagen (Goodpasture antigen), respectively. Seventy-seven percent of the patients had IgM antibodies against the non-Goodpasture glomerular basement membrane (non-GP GBM) in their acute phase serum samples as compared to only 4% seropositivity rate in convalescent phase samples. However, there was no correlation between the non-GP GBM IgM antibody levels and the decrease in GFR as measured by Cr EDTA clearance. None of the patients had antibodies against the Goodpasture antigen. In a control group of 10 patients with influenza and parainfluenza infections serum IgM against non-GP GBM could not be demonstrated, neither in acute phase nor in convalescent phase sera. The possible pathophysiological implications of these findings are discussed. PMID- 9342484 TI - The influence of apoprotein epsilon 2 homozygosity on nephrotic hyperlipidemia. AB - In the normal population, the usual effect of the epsilon 2 allele is to decrease plasma cholesterol and to increase plasma triglyceride. We report here the association of nephrotic syndrome and the apo epsilon 2 epsilon 2 genotype in which we observed a hyperlipidemia characterized by very low levels of lipoprotein lipase activity, chylomicronemia, high levels of plasma apo B, C III, E and lipoprotein(a), very low levels of high density lipoprotein cholesterol and concentrations of cholesterol and triglyceride that are higher than expected in all the other lipoprotein fractions. When proteinuria was partially resolved and plasma albumin levels returned to normal, a residual type III hyperlipidemia was still present. These findings suggest that the combination of apo epsilon 2 homozygosity and massive proteinuria may cause considerable changes in the clearance of triglyceride rich particles probably mediated by the almost complete absence of lipolytic enzymes and a low interaction of lipoproteins with specific receptors. The apo E genotype should be investigated in nephrotic patients with chylomicronemia. PMID- 9342485 TI - Influence of cigarette-smoking on the progression of clinical diabetic nephropathy in type 2 diabetic patients. AB - Cigarette smoking was known to promote the progression of diabetic nephropathy in patients with type 1 diabetes, but its influence on the course of diabetic nephropathy in patients with type 2 diabetes had not been previously established. In a prospective follow-up study we therefore compared the progression of nephropathy in type 2 diabetic patients with or without tobacco consumption. Initiation of dialysis treatment or death of the patient were the end points of the study. 36 patients with type 2 diabetes complicated with diabetic nephropathy were included in the study, 16 smoked and 20 did not. The main outcome measures were proteinuria, arterial blood pressure, HbAlc, serum-creatinine and creatinine clearance, which were controlled at least every six months. In the smoking diabetic patients the mean (SD) creatinine-clearance decreased from 82 +/- 10 to 10 +/- 6 ml/min/1.73 m2 over a period of 62 +/- 21 months. The rate of decline of the creatinine-clearance was 1.24 +/- 0.34 ml/min/month. In the non-smoking patients the creatinine-clearance decreased from 79 +/- 8 to 9 +/- 3 ml/min/1.73 m2 within 79 +/- 27 months. The rate of decline in the creatinine-clearance was 0.99 +/- 0.35 ml/min/month (p < 0.025). HbAlc, systolic and diastolic blood pressure as well as serum cholesterol and triglycerides were not significantly different in both patient groups. Therefore, we conclude that cigarette smoking promotes the progression of diabetic nephropathy in patients with type 2 diabetes, just as it is known in type 1 diabetic patients. PMID- 9342486 TI - Nephelometry in the clinical assessment of glomerular proteinuria and tubular function in diabetic nephropathy. AB - Urinary excretion rate and total clearances of albumin, IgG, IgA and alpha 1 microglobulin, together with selectivity index and proteinuria, were determined by computerized nephelometry in 187 IDDM and NIDDM diabetic out-patients and in 39 healthy subjects in order to perform a prompt clinical assessment of diabetic nephropathy. Significant correlations between nephelometric and RIA procedures were demonstrated for the urinary excretion of albumin (p < 0.001) and total IgG (p < 0.001) in diabetic patients and healthy subjects. Nephelometry allowed us to classify diabetic patients in different stages of nephropathy: non nephropathic, normoalbuminuric with hyperfiltration, with incipient (microalbuminuric) and overt nephropathy (macroalbuminuric). Thirty consecutive subjects were analyzed within 1 h from the beginning of the procedure. A normal tubular function was demonstrated in non nephropathic, hyperfiltering and in 34% of microalbuminuric diabetic patients. On the contrary, in 66% of microalbuminuric and in 93% of macroalbuminuric patients alpha 1-microglobulin urinary levels were found above the upper normal limit. Urinary excretion of IgA was significantly increased only in macroalbuminuric diabetic patients (p < 0.001); this marker might therefore characterise the stage of overt nephropathy. Computerized nephelometry can be considered as a prompt, reproducible and high sensitive approach in the clinical evaluation of proteinuria and tubular function in diabetic renal disease. PMID- 9342487 TI - Pretransplant renal dysfunction predicts poorer outcome in liver transplantation. AB - The postoperative courses of 115 liver transplant recipients were reviewed to monitor for outcomes of acute renal failure and mortality. An analysis of baseline (preoperative) variables with particular attention to baseline renal function was accomplished to establish predictive variables for a complicated postoperative course. Acute renal failure requiring dialysis occurred in 27 cases (23%) and was associated with a prolonged ICU stay, greater infectious complications, greater hospital charges and a high mortality rate (46 +/- 11% vs. 9 +/- 3%) as compared to patients who did not experience acute renal failure. Death occurred in 20 patients (17%) overall prior to discharge. In order to assess the contribution of renal function, the population was divided arbitrarily into two groups based on preoperative serum creatinine. Group 1 (n = 50) had a preoperative serum creatinine < 1.0 mg/dl (mean +/- SD = 2.2 +/- 0.2 mg/dl) and Group 2 (n = 65) had a preoperative serum creatinine < or = 1.0 mg/dl (0.7 +/- 0.1 mg/dl). The groups experienced similar operative courses. Group 1 patients experienced significantly longer ICU stays (18 +/- 3 vs. 10 +/- 2 days), higher rates of acute renal failure requiring dialysis (52 +/- 7 vs. 5 +/- 2%), higher hospital charges (231,454 +/- 17,088 vs. 178,755 +/- 14,744 $, US) and a greatly increased mortality rate (32 +/- 1 vs. 6 +/- 1%), as compared to Group 2 patients. A multifactorial regression analysis demonstrated that of all pretransplant factors analyzed, elevation in the serum creatinine was significantly associated and was the strongest predictor of both outcomes: acute renal failure requiring dialysis (ROC = 0.89) and death (ROC = 0.83). The presence or absence of hepatorenal syndrome did not influence the results of this analysis. This study demonstrates that cirrhotic patients with renal dysfunction, as indicated by an elevated serum creatinine, experience a poor surgical outcome following liver transplantation. These patients may require special attention in the perioperative period. Alternatively, these data may influence the selection of ideal candidates for liver transplantation, where scarce resources need to be applied appropriately. PMID- 9342488 TI - Immunoglobulin G subclasses and susceptibility to allosensitization in humans. AB - Understanding the cellular basis of allosensitization is important because persistence of cytotoxic alloreactive antibodies significantly decreases the chances for receiving a second kidney graft and has a detrimental effect on graft survival rates. In this study, serum levels of IgG subclasses were compared within three groups of uremic patients with different levels of allosensitization (panel reactive antibody level > or = 70%, 10-65% and < or = 10%). All the non sensitized patients had already lost at least one graft indicating resistance to allosensitization by the previous graft. In addition, the in vitro T-cell proliferation and immunoglobulin M and G subclass production were studied after activation by pokeweed mitogen and alloantigens. The patients' demographics were comparable. The results show that all serum IgG subclass levels in the three groups were comparable and within the range of normal control. Similarly, T-cell proliferation and the in vitro production of IgM was not significantly different. The lymphocytotoxic activity present in each IgG subclass was not associated with an increase in the respective serum subclass level or the in vitro production of the same subclass in the sensitized patients. The data indicate that humoral immunity, as reflected by subclass immunoglobulin levels, is, in fact, normal, in the three groups and that sustenance of cytotoxic antibody production reflects a specific immune response controlled by factors other than intrinsic B-cell abnormality. PMID- 9342489 TI - Nephrologists' subjective attitudes towards end-of-life issues and the conduct of terminal care. AB - Decisions which determine the duration and outcome of terminal care should be influenced by patient autonomy. Studies suggest, however, that end-of-life decision-making is more complex than a single principle and that physicians may be responsible for selected aspects of terminal care independent of patient choice. To study how nephrologists' perceptions toward end-of-life issues may affect decision-making, we anonymously surveyed 125 of them. The study employed the straightforward terminology of "hastening death" rather than adopting the ambiguous term "euthanasia" or the narrow term "assisted suicide." Subjective physician profiles demonstrated that nephrologists who are less comfortable with dying patients were significantly less likely to report that they omitted life prolonging measures (p = 0.02) and more likely to report that they would not initiate measures in order to hasten death even were it legal (p = 0.04). Ninety eight percent of nephrologists reported omissions in terminal care with patient knowledge and 80% without patient knowledge. In contrast, forty-three percent of the nephrologists said that were it to become legal to initiate measures in order to hasten death, they would "never" do so. The ethical framework utilized for discontinuation of dialysis decisions incorporated medical benefit (cancer as criterion, 48%; multisystem complications, 84%; dementia 79%) and quality of life criteria. Twenty-five percent of nephrologists admitted difficulty with advance directives if the directives clashed with heir beliefs. ESRD end-of-life decision making in the USA may be altered by the subjective characteristics of nephrologists. In particular, nephrologists' level of discomfort with patient mortality is linked with their reported management of terminal patients. PMID- 9342491 TI - Increased endothelin-1 content in the platelets of hemodialysed patients. AB - Patients undergoing hemodialysis, present platelets (PLTs) with physiological dysfunction. Aggregated PLTs stimulate endothelin-1 (ET-1) secretion from endothelial cells. In turn, ET-1 abolishes PLT aggregation. The aim of this study was to determine any presence of ET-1 in the PLTs of hemodialysed patients and to compare its levels with normal subjects. Platelets, isolated from hemodialysed patients, revealed lower aggregation (76 +/- 13%) compared with healthy subjects (96 +/- 2%). Plasma ET-1 was increased in hemodialysed patients 23.5 +/- 3.2 fmol/ml, vs. 10.9 +/- 1.6 fmol/ml in normal subjects. Immunoreactive endothelin-1 was detected in the platelets of both groups. The intraplatelet ET-1 of hemodialysed patients was 13.8 +/- 3.1 fmol/mg protein, vs. 7.9 +/- 1.3 fmol/mg protein in normal individuals. Total RNA was isolated from platelets and reverse transcriptase-mediated polymerase chain reaction (RT-PCR) revealed no presence of the preproET-1 mRNA in either normal or hemodialysed platelets. This suggests that ET-1 is internalized from the plasma. In summary, platelets contain but do not express ET-1. The increased levels of ET-1 in the platelets of the hemodialysed patients may be partly responsible for their lower aggregation. PMID- 9342490 TI - Calcium salts of keto-amino acids, a phosphate binder alternative for patients on CAPD. AB - Control of hyperphosphoremia is crucial to the prevention of secondary hyperparathyroidism. Calcium salts of keto-amino acids (KAA) were employed as phosphate binders in hemodialysis patients. We wanted to assess the efficacy of these substances as quelating agents in patients under continuous ambulatory peritoneal dialysis (CAPD). Also, as an amino acid supplement, we determined their possible effect on some parameters related to nutritional status. We studied 13 patients (7 M; 6 F) with a mean age of 45.2 +/- 17 years and a mean time on CAPD of 18.4 +/- 11.4 months. None had severe secondary hyperparathyroidism and/or clinically relevant aluminium intoxication. They were not receiving calcitriol and none were using low-calcium peritoneal dialysis fluids. All were under aluminum hydroxide (AlOH3) treatment and 8 patients also received calcium carbonate. These quelating agents were withdrawn and after 21 days (wash-out period) KAA were initiated. We analyzed serum levels of bone metabolism parameters (calcium, phosphate, osteocalcin [OC], intact parathyroid hormone [iPTH], alkaline phosphatase [AP]) and nutritional parameters (total protein, albumin, pre-albumin, transferrin) in four periods: (A) during AlOH3; (B) immediately after the washout period; (C) after 1.5 months; and (D) after 3 months of KAA therapy. In 5 patients serum aluminum level was also measured in periods (A) and (D). The serum phosphate level at period (B) was significantly higher than in other periods. After 3 months of treatment phosphate levels decreased significantly (A = 1.77 +/- 0.3 mmol/l vs D = 1.48 +/- 0.2; p < 0.05). Serum calcium levels increased, while iPTH and OC decreased (p = ns). AP remained stable during the study. All nutritional parameters increased at the end of the study (p = ns). Calcium salts of keto-amino acids showed to be an effective alternative to aluminum-containing phosphate binders. They were well tolerated, without relevant side-effects. These compounds could also represent an additional source of oral amino acid supplementation with improvement of nutritional status. PMID- 9342492 TI - Large, rapid skeletal changes after parathyroidectomy. AB - We report dramatic increases in spine and hip bone mineral density six weeks following parathyroidectomy in renal patients. We have previously reported a cross-sectional association between parathyroidectomy and higher axial bone mineral density in dialysis patients. Large axial increases in bone mineral density after surgery for primary hyperparathyroidism have been recorded by others at one year postoperatively. Bone mineral density was recorded before and six weeks after parathyroidectomy. Scans were performed at the lumbar spine, hip and non-dominant radius. Values were expressed as Z-scores (standard deviations from the mean of an age- and gender-matched reference population). Large increases in lumbar were spine and femoral neck bone mineral density seen at six weeks post parathyroidectomy. Median increases were 0.57 (p = 0.006) and 0.26 (p = 0.039) Z-score units for these sites, respectively. Individual six-week increases were as large as 1.3 Z-score units at the spine and 1.0 Z-score units at the femoral neck. No significant cohort change was detected at the forearm but individual forearm changes were highly variable. Several mechanisms to explain these large rapid increases can be postulated. These include: mineralization of osteoid and/or contraction of the remodeling space. The changes illustrate the extent to which the skeleton is capable of rapid and profound change in mineral content. Our findings emphasize that the skeleton is a heterogeneous organ and that bone density should be measured at multiple skeletal sites. PMID- 9342493 TI - Massive uncomplicated vascular immune complex deposits in the kidney of a patient with systemic lupus erythematosus. AB - The case of a patient with systemic lupus erythematosus (SLE) is reported which was accompanied by renal dysfunction and massive vascular immune deposits in the kidney without active glomerular lesions. The renal biopsy showed arterioles and small arteries with circumferential periodic acid-Schiff (PAS) and Masson trichrome-positive homogenous material in the subendothelial area in the absence of thrombotic, necrotizing or inflammatory lesions. Immunofluorescence and electron microscopy examination demonstrated immune deposits in the vascular walls. Glomeruli showed only minor abnormalities with a trend to collapse. There was no improvement in renal dysfunction over a 4-year period until the patient's death, despite steroid therapy producing a decrease in disease activity. The autopsy showed similar vascular changes to those seen in the biopsy, however; glomeruli were either sclerotic or showed a trend to collapse. Massive uncomplicated vascular immune complex deposition without active glomerular lesions is rare. The present case indicates that this type of lupus vasculopathy may be a prognostic factor for the loss of renal function in SLE mediated by hemodynamic glomerular injury. PMID- 9342494 TI - Urinary beta-glucuronidase excretion in insulin-dependent diabetes mellitus: relation with diabetic nephropathy. PMID- 9342495 TI - Reply to Shoker et al. PMID- 9342496 TI - Lupus nephritis in a patient with idiopathic thrombocytopenic purpura diagnosed 11 years earlier. PMID- 9342497 TI - Sodium ferric gluconate and iron requirements in hemodialysis patients. PMID- 9342498 TI - Reply to Vleming et al. PMID- 9342499 TI - "Beer potomania" in a non-beer drinker. PMID- 9342500 TI - Renal involvement in diffuse fasciitis with eosinophilia. PMID- 9342501 TI - Clinical pharmacokinetics of irinotecan. AB - This article reviews the clinical pharmacokinetics of a water-soluble analogue of camptothecin, irinotecan [CPT-11 or 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxy-camptoth eci n]. Irinotecan, and its more potent metabolite SN-38 (7- ethyl-10-hydroxy-camptothecin), interfere with mammalian DNA topoisomerase I and cancer cell death appears to result from DNA strand breaks caused by the formation of cleavable complexes. The main clinical adverse effects of irinotecan therapy are neutropenia and diarrhoea. Irinotecan has shown activity in leukaemia, lymphoma and the following cancer sites: colorectum, lung, ovary, cervix, pancreas, stomach and breast. Following the intravenous administration of irinotecan at 100 to 350 mg/m2, mean maximum irinotecan plasma concentrations are within the 1 to 10 mg/L range. Plasma concentrations can be described using a 2- or 3-compartment model with a mean terminal half-life ranging from 5 to 27 hours. The volume of distribution at steady-state (Vss) ranges from 136 to 255 L/m2, and the total body clearance is 8 to 21 L/h/m2. Irinotecan is 65% bound to plasma proteins. The areas under the plasma concentration-time curve (AUC) of both irinotecan and SN-38 increase proportionally to the administered dose, although interpatient variability is important. SN-38 levels achieved in humans are about 100-fold lower than corresponding irinotecan concentrations, but these concentrations are potentially important as SN-38 is 100- to 1000-fold more cytotoxic than the parent compound. SN-38 is 95% bound to plasma proteins. Maximum concentrations of SN-38 are reached about 1 hour after the beginning of a short intravenous infusion. SN-38 plasma decay follows closely that of the parent compound with an apparent terminal half-life ranging from 6 to 30 hours. In human plasma at equilibrium, the irinotecan lactone form accounts for 25 to 30% of the total and SN-38 lactone for 50 to 64%. Irinotecan is extensively metabolised in the liver. The bipiperidinocarbonylxy group of irinotecan is first removed by hydrolysis to yield the corresponding carboxylic acid and SN-38 by carboxyesterase. SN-38 can be converted into SN-38 glucuronide by hepatic UDP glucuronyltransferase. Another recently identified metabolite is 7-ethyl-10-[4-N (5-aminopentanoic acid)-1-piperidino]-carbonyloxy-camptothecin (APC). This metabolite is a weak inhibitor of KB cell growth and a poor inducer of topoisomerase I DNA-cleavable complexes (100-fold less potent than SN-38). Numerous other unidentified metabolites have been detected in bile and urine. The mean 24-hour irinotecan urinary excretion represents 17 to 25% of the administered dose. Recovery of SN-38 and its glucuronide in urine is low and represents 1 to 3% of the irinotecan dose. Cumulative biliary excretion is 25% for irinotecan, 2% for SN-38 glucuronide and about 1% for SN-38. The pharmacokinetics of irinotecan and SN-38 are not influenced by prior exposure to the parent drug. The AUC of irinotecan and SN-38 correlate significantly with leuco-neutropenia and sometimes with the intensity of diarrhoea. Certain hepatic function parameters have been correlated negatively with irinotecan total body clearance. It was noted that most tumour responses were observed at the highest doses administered in phase I trials, which indicates a dose-response relationship with this drug. In the future, these pharmacokinetic-pharmacodynamic relationships will undoubtedly prove useful in minimising the toxicity and maximise the likelihood of tumour response in patients. PMID- 9342503 TI - Clinical pharmacokinetics of stavudine. AB - Stavudine (d4T) is a pyrimidine nucleoside analogue used in the treatment of human immunodeficiency virus (HIV) infection. It inhibits viral reverse transcriptase as do zidovudine (AZT), didanosine (ddI), zalcitabine (ddC) and lamivudine (3TC), which comprise the family of nucleoside HIV-reverse transcriptase inhibitors. Stavudine is currently approved by the US Food and Drug Administration for the treatment of patients who have become intolerant to or have failed to response to zidovudine, didanosine or zalcitabine therapy. Oral administration of stavudine results in maximal concentrations within 2 hours and increases linearly as doses increase. The absolute oral bioavailability is high, approaching 100%. There is evidence to suggest that stavudine does not accumulate in the plasma. It distributes into total body water and appears to enter cells by non-facilitated diffusion. Penetration into the cerebrospinal fluid occurs, as does the transfer of the drug across human placental tissue. Stavudine is cleared quickly by both renal and nonrenal processes. The pharmacokinetic properties of stavudine in children are similar to those of adults. The pharmacokinetic parameters of stavudine were not affected by simultaneous administration of didanosine. It appears that stavudine at doses < 2 mg/kg/day is most efficient at increasing CD4 + cell numbers. While stavudine is reported to be less cytotoxic than zidovudine, the principal toxicity in humans is peripheral neuropathy and appears to be related to daily, but not cumulative, doses. PMID- 9342502 TI - Clinical pharmacokinetics of nefazodone. AB - Nefazodone is a new antidepressant drug, chemically unrelated to the tricyclic, tetracyclic or selective serotonin uptake inhibitors. Nefazodone blocks the serotonin 5-HT2 receptors and reversibly inhibits serotonin reuptake in vivo. Nefazodone is completely and rapidly absorbed after oral administration with a peak plasma concentration observed within 2 hours of administration. Nefazodone undergoes significant first-pass metabolism resulting in an oral bioavailability of approximately 20%. Although there is an 18% increase in nefazodone bioavailability with food, this increase is not clinically significant and nefazodone can be administered without regard to meals. Three pharmacologically active nefazodone metabolites have been identified: hydroxy-nefazodone, triazoledione and m-chlorophenylpiperazine (mCPP). The pharmacokinetics of nefazodone are nonlinear. The increase in plasma concentrations of nefazodone are greater than would be expected if they were proportional to increases in dose. Steady-state plasma concentrations of nefazodone are attained within 4 days of the commencement of administration. The pharmacokinetics of nefazodone are not appreciably altered in patients with renal or mild-to-moderate hepatic impairment. However, nefazodone plasma concentrations are increased in severe hepatic impairment and in the elderly, especially in elderly females. Lower doses of nefazodone may be necessary in these groups. Nefazodone is a weak inhibitor of cytochrome P450 (CYP) 2D6 and does not inhibit CYP1A2. It is not anticipated that nefazodone will interact with drugs cleared by these isozymes. Indeed, nefazodone did not affect the pharmacokinetics of theophylline, a compound cleared by CYP1A2. Nefazodone is metabolised by and inhibits CYP3A4. Clinically significant interactions have been observed between nefazodone and the benzodiazepines triazolam and alprazolam, cyclosporin and carbamazepine. The potential for a clinically significant interaction between nefazodone and other drugs cleared by CYP3A4 (e.g. terfenadine) should be considered before the coadministration of these compounds. There was an increase in haloperidol plasma concentrations when coadministered with nefazodone; nefazodone pharmacokinetics were not affected after coadministration. No clinically significant interaction was observed when nefazodone was administered with lorazepam, lithium, alcohol, cimetidine, warfarin, theophylline or propranolol. PMID- 9342506 TI - Epidemiology of nonmelanoma skin cancer. AB - In summary, the cause of NMSC is multifactorial and complex. The interaction among environmental and lifestyle factors along with host factors determines the development and progression of NMSC. Despite the relatively benign nature of NMSC, rising incidence rates in an aging population will result in increased medical costs, morbidity, and mortality. Frequent exposure to UVR, fair skin, and reduced immunity, possibly related to aging, are the major risk factors. Because UVR exposure is responsible for most of the NMSC cases, the key to reducing the number of skin cancers is through prevention, mainly by reducing exposure to sunlight, and further exploration of possible chemopreventive agents. To understand better the complex causes of NMSC and to evaluate prevention strategies, further population-based research is necessary. PMID- 9342504 TI - Pharmacokinetic considerations of new insulin formulations and routes of administration. AB - There is a continuing search for improved insulin formulations in order to imitate as closely as possible the physiological pattern of insulin secretion, and thereby to minimise the complications of diabetes mellitus. The major advances achieved to date are in the area of human insulin analogue synthesis resulting from the introduction of recombinant DNA techniques, and in improved delivery systems that utilise noninvasive or minimally invasive modes of administration. To accommodate postprandial hyperglycaemia, monomeric insulin formulations have been developed, of which insulin lispro (the Lys-Pro analogue) is already approved for clinical use. These formulations have a rapid rate of absorption and, therefore, have to be administered at meal time (unlike the previous short-acting formulations). Their residence time is also about 2-fold shorter than regular human insulin; this minimises the risk of the excessive hypoglycaemic effect that characterises regular human insulin formulations. Certain proinsulin formulations with hepatoselective activity have been developed but were found to be poorly tolerated. The newer proinsulin molecules do not show hepato-selective properties. In order to generate basal insulin levels, peakless long-acting formulations have been developed, including: a soluble formulation (which upon subcutaneous administration, produces a 'depot-like' sustained release mechanism), albumin-bound insulin and cobalt-insulin hexamer formulations. To improve patient compliance the 'pen' device was developed for subcutaneous injections. Programmable infusion pumps were developed to avoid repetitive subcutaneous injections. Great efforts have been made in searching for noninvasive modes of insulin administration that will avoid the need for parenteral administration. These include: oral, colonic, rectal, nasal, ocular, buccal, pulmonary, uterine and transdermal routes of administration. Various enhancers have been tested to increase the bioavailability of each route. At present these alternative routes do not provide clinically relevant substitutes for the subcutaneous mode of administration. In conclusion, although the newer formulations provide certain advantages, there is still much to be done to further facilitate and improve insulin therapy. PMID- 9342507 TI - Immune response associated with nonmelanoma skin cancer. AB - It is now clear that UV radiation causes nonmelanoma skin cancer in at least two ways: by causing permanent changes in the genetic code and by preventing immunologic recognition of mutant cells. These are interacting rather than separate mechanisms. Damage to DNA results in disregulation of cellular proliferation and initiates immune suppression by stimulating the production of suppressive cytokines. These cytokines contribute to the loss of immunosurveillance. Ultraviolet radiation has both local and systemic immunosuppressive effects. Locally, it depletes and alters antigen-presenting LC at the site of UV irradiation. Systemic suppression results when Ts cells are induced, by altered LC, by inflammatory macrophages that enter the skin following UV irradiation, or by the action of cytokines. Damage to DNA appears to be one of the triggering events in inducing systemic immunosuppression via the release of immunosuppressive cytokines and mediators. Immunologic approaches to treating skin cancers so far have concentrated on nonspecifically stimulating immune cells that infiltrate these tumors, but induction of specific immune responses against these tumors with antitumor vaccines has received little attention as yet. Preventive measures include sun avoidance and the use of sunscreens to prevent DNA damage by UV light. Future strategies may employ means to reverse UV-induced immunosuppression by using anti-inflammatory agents, biologicals that accelerate DNA repair or prevent the generation of immunosuppressive cytokines, and specific immunotherapy with tumor antigens. New approaches for studying the immunology of human skin cancers are needed to accelerate progress in this field. PMID- 9342508 TI - Congenital syndromes associated with nonmelanoma skin cancer. AB - Syndromes associated with nonmelanoma skin cancer are part of a group of inherited disorders characterized by a wide variety of clinical manifestations including palmar and plantar pitting, calcification of the dura, jaw cysts, and skeletal abnormalities. The syndromes have in common skin lesions progressing to basal or squamous cell cancers. In addition to skin tumors, bone and visceral tumors are common. Early diagnosis, genetic counseling, and treatment of skin lesions are appropriate tools for the management of patients with these syndromes. Surgical procedures generally involve total resection combined with reconstructive efforts. PMID- 9342505 TI - Clinical pharmacokinetic considerations in the elderly. An update. AB - There are a number of areas in which advances have been made over the last few years in the area of pharmacokinetics in the elderly. There is increasing understanding of the diversity of cytochrome P450s (CYP) and the variability of the age-related decline in CYP activity. This has helped to explain some of the interindividual variability in drug metabolism with age. The importance of ethnic differences has emerged, but specific work is needed in this area in the elderly. Differences in the handling of chiral compounds has been reported but as yet no clinically important findings that may lead to a change in clinical practice have emerged. The emerging importance of extrahepatic drug metabolism, especially in the intestine, has added a new complexity to our understanding of pharmacokinetics. The issue of frailty is also discussed in this article. Whether it will be of value at the bedside has yet to emerge. Nonetheless, as a concept, recent data has supported its potential use to define those more at risk of clinically meaningful pharmacokinetic alterations. Other advances have included the appreciation that selectivity in induction and inhibition in the elderly are due to the existence of multiple CYP forms. Similarly, the role of these various enzymes in disease is also improving our clinical understanding, as exemplified in Parkinson's disease. PMID- 9342509 TI - Prevention of nonmelanoma skin cancer. Standard and investigative approaches. AB - Today, in many fields of medicine, the focus is on prevention. This discussion highlights the salient features of sunscreens, protective clothing, and education, as they relate to the hazards and potential sequelae of sun exposure and tanning parlors. It also includes information on the status of clinical and laboratory investigation in the development of agents to prevent skin cancer in high-risk individuals. PMID- 9342510 TI - Assessment and surgical treatment of basal cell skin cancer. AB - The conspicuously high rate of BCC in the population mandates that clinicians have a clear understanding of the tumor's patterns of presentation and behavior. Goals of any form of treatment include establishment of the proper diagnosis and eradication of the lesion with a reasonable aesthetic result. PMID- 9342511 TI - Squamous cell and adnexal carcinomas of the skin. AB - The diagnosis and treatment of adnexal cancers continues to pose a challenge to a wide range of clinicians. The diseases are a diverse lot, owing to the wide range of skin structures and large surface area. Early recognition and treatment are key to improved outcomes. Education of the patients as to their role in their care, especially early detection, is also of crucial importance. Further study may yield information to improve diagnosis and treatment. PMID- 9342512 TI - Mohs' micrographic surgery for nonmelanoma skin cancers. AB - The incidence of NMSC is increasing at a phenomenal rate, with a current estimated annual incidence in the United States of more than 600,000 cases. It is vitally important for all surgeons to become familiar with all modalities, surgical as well as nonsurgical, that are used to treat these skin cancers. This article was designed to inform the reader about the surgical technique called MMS (Figs. 9 and 10), and to present the facts regarding its historical evolution, actual technique, indications, and limitations. MMS has been modified and refined over a period spanning 60 years. During this time, tens of thousands of tumors have been treated, and the literature supports the fact that MMS provides the highest possible cure rates for the treatment of NMSCs. PMID- 9342513 TI - Radiation therapy for nonmelanoma skin carcinomas. AB - Radiotherapy plays an important role in the management of appropriately selected skin carcinomas. Tumors of the eyelid, nose, and ear can be treated successfully with preservation of adjacent normal tissues. Tumors located in the embryologic fusion planes of the face can be irradiated with wide margins, either as primary or postoperative therapy. Radiotherapy has a significant role in the postoperative setting for patients with high-risk features in their pathologic specimens. Patients with NECS (Merkel cell carcinoma) have a high recurrence rate after surgical excision; these patients should undergo radiotherapy after resection of the clinically evident disease. PMID- 9342515 TI - Local flap reconstruction of defects after excision of nonmelanoma skin cancer. AB - Reconstruction of facial defects poses the interesting challenge of finding the most satisfactory flap both aesthetically and functionally. It requires not just a knowledge of the flap, but an ability to think and plan in three dimensions. Not all individuals possess this; thus, what is obvious and simple to one surgeon, poses a great and worrisome problem for another. This can be made easier by considering certain rules. First, excise the lesion properly and then think about the reconstruction. The presence of the defect crystallizes the thought process. Next, consider the topographic and functional anatomy of the face, the differences in skin color, presence of hair, and the "idea" lines for position of scars. The flap options are rotation, transposition, and advancement. The latter, on occasions, may be as an island. These movements must be fitted into the aforementioned requirements. In this way the best choice usually is reached. Remember, there is always a way out using a skin graft or tissue expansion. PMID- 9342514 TI - Advanced and recurrent nonmelanoma skin cancer. AB - Advanced skin cancer presents a substantial challenge to the surgeon, who must incorporate sound oncologic principles and carefully considered reconstruction into the treatment plan. Using an anatomical format, this article discusses the treatment plans for advanced skin cancer and outlines various pitfalls. The appropriateness and advisability of other modalities are also discussed. PMID- 9342517 TI - Management of nonmelanoma skin tumors of the hand. AB - A wide variety of tumors can afflict the hand. Nonmelanoma skin tumors are common and usually benign. The most common malignant tumor of the hand is a SCC. Whether benign or malignant, these need neoplasms and are often initially symptomatic, presenting as an area of discoloration or excoriation. The tumor mass may become disfiguring or impair function. It behooves all hand surgeons to be aware of these nonmelanoma skin tumors and to secure prompt recognition and early treatment, thereby optimizing long-term results. PMID- 9342516 TI - Prosthetic rehabilitation of patients with advanced nonmelanoma skin cancer. AB - Attention to detail at all stages of treatment can ensure a successful prosthetic rehabilitation. This detail must be considered a priority at presurgery, surgery, and at every stage in fabricating the prosthesis. Teamwork between the surgeon and maxillofacial prosthodontist will ensure an optimal surgical preparation and definitive prosthesis. Evidence of interaction among team members most certainly can be encouraging to the patient. It is important that during the prosthetic phase of treatment, attention be made in terms of tissue assessment, impression making, sculpting, mold fabrication, familiarity with materials, appreciation of color, delivery of instructions, and patient education, which will ensure a satisfactory outcome. With the desire, determination, and encouragement from the restorative team to make the most of this artificial replacement, a patient can have a higher quality of life and a more normalized lifestyle. PMID- 9342518 TI - Microvascular reconstruction for cutaneous defects. AB - Advanced skin cancer of the head and neck is best managed by a multidisciplinary approach. The goal is to control the malignancy while maximizing function and aesthetics, and hence the quality of life, after therapy. As a member of the patient care team, the reconstructive surgeon brings specialized expertise in modern techniques of tissue transfer to provide the optimal restoration for patients following resection of the tumor. Microvascular surgery represents the current technical state of the art for reconstruction in these patients with advanced lesions. PMID- 9342519 TI - The avian embryo as a model in developmental studies: chimeras and in vitro clonal analysis. AB - The avian embryo is a model in which techniques of experimental embryology and cellular and molecular biology can converge to address fundamental questions of development biology. The first part of the chapter describes two examples of transplantation and cell labeling experiments performed in ovo. Thanks to the distinctive histologic and immunocytochemical characteristics of quail and chick cells, the migration and development of definite cells are followed in suitably constructed chimeric quail-chick embryos. Isotopic transplantations of neural tube portions between quail and chick, combined with in situ hybridization with a nucleic probe specific for a quail oligodendrocyte marker, allowed study of the origin and migration of oligodendroblasts in the spinal cord. Heterotopic transplantations of rhombomeres were performed to establish the degree of plasticity of these segments of the hindbrain regarding Hox gene expression, which was revealed by labeling with chick-specific nucleic probes. The second part describes in vitro cell cloning experiments devised to investigate cell lineage segregation and diversification during development of the NC. An original cloning procedure and optimal culture conditions permitted analysis of the developmental potentials of individual NC cells taken at definite migration stages. The results revealed a striking heterogeneity of the crest cell population, which appeared to be composed of precursors at different states of determination. Clonal cultures also provide a means to identify subsets of cells that are the target of environmental factors and to understand how extrinsic signals influence the development of responsive cells. PMID- 9342520 TI - Inhibition of gene expression by antisense oligonucleotides in chick embryos in vitro and in vivo. PMID- 9342521 TI - Lineage analysis using retroviral vectors. PMID- 9342522 TI - Use of dominant negative constructs to modulate gene expression. PMID- 9342523 TI - The use of embryonic stem cells for the genetic manipulation of the mouse. PMID- 9342524 TI - Organoculture of otic vesicle and ganglion. PMID- 9342525 TI - Organoculture of the chick embryonic neuroretina. PMID- 9342526 TI - Embryonic explant and slice preparations for studies of cell migration and axon guidance. PMID- 9342527 TI - Culture of avian sympathetic neurons. PMID- 9342528 TI - Analysis of gene expression in cultured primary neurons. PMID- 9342529 TI - Selective aggregation assays for embryonic brain cells and cell lines. PMID- 9342530 TI - Flow cytometric analysis of whole organs and embryos. PMID- 9342531 TI - Detection of multiple gene products simultaneously by in situ hybridization and immunohistochemistry in whole mounts of avian embryos. PMID- 9342532 TI - Cloning of genes from single neurons. PMID- 9342533 TI - Methods for detecting and quantifying apoptosis. PMID- 9342534 TI - Methods in Drosophila cell cycle biology. PMID- 9342535 TI - Single central nervous system neurons in culture. PMID- 9342536 TI - Patch-clamp recordings from Drosophila presynaptic terminals. PMID- 9342537 TI - Physiology, pathology and pharmacology of insulin secretion: recent acquisitions. AB - Recent acquisitions concerning the physiology, pathology and pharmacology of insulin secretion are reviewed. In terms of physiology, emphasis is placed on new information concerning the role of glucokinase and the identity of coupling factors in the process of glucose-stimulated insulin release. Pathological considerations concern mainly the possible participation of an inherited or acquired defect of FAD-linked mitochondrial glycerophosphate dehydrogenase in the impairment of insulin release in non-insulin-dependent diabetes. Although experimental approaches to correct such a site-specific defect have so far been unsuccessful, new therapeutic tools, especially the esters of certain nutrients, may soon be available for stimulation of proinsulin biosynthesis as well as insulin release in the diseased B cell. PMID- 9342538 TI - Is leptin the link between obesity and insulin resistance? AB - Leptin is the product of OB gene. This 16 kDa protein is produced by mature adipocytes and is secreted in plasma. Its plasma levels are strongly correlated with adipose mass in rodents as well as in humans. Leptin inhibits food intake, reduces body weight and stimulates energy expenditure. It has been suggested that leptin could be the link between obesity and diabetes. Recent experiments in rodents have shown that leptin expression in adipocytes is also regulated at short-term by hormones and nutrients. Leptin expression increases after food intake and decreases during fasting and diabetes. Insulin and glucocorticoids increase leptin expression, whereas catecholamines, via beta-adrenergic receptors and cAMP, and long-chain fatty acids (and thiazolidinediones), via PPARy, inhibit leptin expression. Leptin is a cytokine that binds to transmembrane receptors similar to the receptors of cytokine family (type IL-6), and transmit their information inside the cell, after dimerisation. A short-form of leptin receptor (with a cytoplasmic domain of 34 amino residues) has been identified in the choroid plexus. This type of receptor should be used for leptin transport across the blood-brain barrier. Then leptin binds to a long-form of leptin receptor in the hypothalamus (with a cytoplasmic domain of 302 amino residues) and decreases the production of neuropeptide Y, a neuromediator of food intake. The long-form of leptin receptor, transmits its information via the Janus Kinases (JAK) who subsequently phosphorylate transcription factors of the STAT family. Intermediary forms of leptin receptor have been identified in other tissues: liver, heart, skeletal muscles, endocrine pancreas. The role of leptin receptors in these tissues remains obscure, but is of considerable interest. Recent studies have shown that leptin inhibits insulin secretion and have anti-insulin effects on liver and adipose tissue. If these effects are confirmed, leptin could play a role similar to TNF alpha and could participate in the insulin-resistance of obesity and type II diabetes. PMID- 9342539 TI - Insulin and the prevention of insulin-dependent diabetes mellitus. AB - Insulin deficiency due to autoimmune destruction of pancreatic beta cells (insulin is an autoantigen) is responsible for insulin-dependent diabetes mellitus. Since 1923, substitutive administration of insulin has been used to treat the disease. Surprisingly, initial usage of insulin is associated with partial resumption of insulin secretion in most patients. This phenomenon is intensified by aggressive insulin therapy. When observed at a late phase of destruction, it has been interpreted as an immunomodulatory effect of insulin which is presumed to act either by masking the target of effector cells in the autoimmune reaction (beta cells at rest because of glycaemic normalisation would expose fewer antigens) or by direct action on autoreactive T lymphocytes (which are rich in insulin receptors). There could also be a direct beneficial effect on anti-apoptotic or pro-regenerative beta cells. Efficient prevention of diabetes has been achieved by administration of parenteral insulin to non-obese diabetic (NOD) mice. Certain sequences of the B chain appear to be responsible for this effect, which seems to be immunomediated. Some preliminary data from the groups of G. Eisenbarth and N. MacLaren have suggested that this effect could be obtained in man by administering small doses of subcutaneous insulin to prediabetic patients. Two trials have been under way since 1994: DPT1 (a non randomised trial concerning children and adults at high risk) in the United States, and EPP-SCIT (a randomised trial concerning children at very high risk) in Europe. Another approach has also been attempted in diabetes as well as other diseases with an organ-specific autoimmune reaction (SEP, PR) i.e. oral administration of an antigen present at the reaction site. A positive effect has been shown by the group of H. Weiner in the NOD mouse in which islet infiltration was reduced and diabetes prevented by "oral tolerisation" with insulin. Oral insulin is easy to use in therapeutic studies and is currently being administered in two trials: DPT1 (prediabetic children and adults at moderate risk) in the United States, and DIOR (recently diabetic children and adults) in France. However, recent experimental data suggest that oral administration of an antigen in certain artificial circumstances can trigger an autoimmune reaction in the animal, which would indicate that due caution should be exercised in trials involving prediabetic patients. PMID- 9342540 TI - Hypoglycaemia unawareness. AB - Hypoglycaemia unawareness, a frequent syndrome in insulin-dependent diabetes mellitus (IDDM), involves a decrease or absence of perception of specific symptoms which normally inform the subject that plasma glucose is decreasing to dangerous levels leading to neuroglycopenia. Without warning symptoms, IDDM patients are unable to take measures (e.g. eating) to prevent severe neuroglycopenia (unconsciousness). As hypoglycaemia unawareness is associated with impaired glucose counterregulation, especially reduced adrenaline responses, it can lead to severe hypoglycaemia. Various studies in animals and humans have indicated that hypoglycaemia unawareness is largely, if not entirely, secondary to increased brain glucose transport due to recurrent or chronic hypoglycaemia. In fact, meticulous prevention of hypoglycaemia largely restores the warning symptoms and adrenaline responses, at least in short-term IDDM. In long-term IDDM, recovery is less complete. Diabetologists and IDDM patients need to be familiar with hypoglycaemia unawareness and how to prevent or treat it. Intensive therapy strictly for normoglycaemia may actually increase the frequency of hypoglycaemia and hypoglycaemia unawareness. However, if intensive therapy is combined with a hypoglycaemia prevention programme, the percentage of HbA1c can be maintained below risk values for the onset or progression of complications, and the frequency of hypoglycaemia can be kept low. Under these conditions, IDDM patients can maintain the warning symptoms and adrenaline response to hypoglycaemia, ensuring a vital backup system for safe intensive therapy of IDDM. PMID- 9342541 TI - Diabetes mellitus, hypertension and ageing: the ionic hypothesis of ageing and cardiovascular-metabolic diseases. AB - Ageing in industrialised societies is associated with an increasing prevalence of hypertension, atherosclerotic vascular diseases, reduced insulin sensitivity and non-insulin-dependent diabetes mellitus (NIDDM). It has been suggested that hyperinsulinaemia/insulin resistance and/or hyperglycaemia could play a role in determining and/or exacerbating the hypertension and vascular disease associated with diabetes mellitus and ageing. Insulin-resistant states, such as essential hypertension and NIDDM, as well as "normal" ageing, are characterised by similar intracellular ionic defects, i.e. accumulation of cytosolic free calcium and depletion of free magnesium. The importance of calcium and magnesium ions in regulating cell functions is well-known. A rise in cellular free calcium and a depletion in cellular magnesium may induce cellular insulin resistance and vasoconstriction. Ionic levels quantitatively predict the extent of elevated blood pressure, fasting blood glucose, HBA1c and hyperinsulinaemic response to oral glucose challenge. We suggest that ionic disturbance might be the missing link responsible for the frequent clinical coexistence of hypertension, atherosclerosis and metabolic disorders. Ageing cells may become more vulnerable to ion disturbances, leading to possible elevation of intracellular free calcium and concurrent magnesium depletion. The "ionic hypothesis" of ageing supposes that an alteration in the cellular mechanisms which maintain the homeostasis of cytosolic calcium concentrations may play a key role in the ageing process, and that a sustained accumulation of cellular calcium and/or the depletion of cellular magnesium may also provide the final common pathway for many ageing associated diseases, including hypertension and NIDDM. PMID- 9342542 TI - Influence of metabolic and genetic factors on tumour necrosis factor-alpha and lymphotoxin-alpha production in insulin-dependent diabetes mellitus. AB - The potential role of tumour necrosis factors (TNFs) in autoimmunity and insulin dependent diabetes mellitus (IDDM) led us to determine in vitro TNF-alpha and lymphotoxin-alpha (LT-alpha, TNF-beta) production in IDDM patients according to TNF polymorphism. LT-alpha production of peripheral blood mononuclear cells (PBMC) was lower in diabetic subjects (m = 0.30 +/- 0.2 ng.10(-6) cells) than controls (m = 0.68 +/- 0.3 ng.10(-6) cells, p < 0.05), and early age-at-onset was correlated with low LT-alpha production (rs = 0.8, p = 0.0006). TNF-alpha production was the same in patients and controls, but patients with HbA1c > or = 8% had a higher TNF-alpha production (m = 3.05 +/- 1.2 ng.10(-6) cells) than those with HbA1c < 8% (m = 1.31 +/- 0.33 ng.10(-6) cells, p < 0.05). A study of the microsatellite TNFa region close to the LTA gene showed that the presence of the TNFa1 allele in HLA-(DR3) subjects was associated with increased risk of IDDM. TNFa1-positive subjects (both patients and controls) also had lower LT alpha production than other subjects. These results indicate that low LT-alpha production is an additional risk factor for IDDM and that poor glycaemic control in patients is associated with enhanced PBMC TNF-alpha production which causes an imbalance between TNF-alpha and LT-alpha production in IDDM patient. PMID- 9342543 TI - Increased prevalence of thyroid autoantibodies and subclinical thyroid failure in relatives of patients with overt endocrine disease-associated diabetes but not type 1 diabetes alone. AB - The purpose of this study was to determine the prevalence of thyroperoxidase (TPO) and thyroglobulin (Tg) antibodies, using a sensitive and specific radioimmunoassay method in a large cohort of 254 first-degree relatives of Type 1 diabetic patients with or without other autoimmune endocrinopathy, and to evaluate the predictive value of thyroid antibodies for impaired thyroid function in these groups. TPO and Tg antibodies were found at similar frequencies (12%) in the 254 relatives, and both antibodies were present in 23 cases (9%). Seven subjects displayed subclinical thyroid dysfunction without an abnormal free T4 level. Among first-degree relatives of probands with Type 1 diabetes alone, TPO or Tg antibodies were found in 8 subjects (6%), including 6 with both antibodies. The prevalence of TPO antibodies was significantly greater among relatives of TPO positive than TPO-negative probands (p < 0.01). In relatives of diabetic patients with other endocrinopathy, frequencies of TPO (20%), Tg (19%) and a combination of both antibodies (15%) were significantly higher than in relatives of Type 1 diabetic patients without endocrinopathy (p < 0.001). TSH levels were abnormal in only one relative of the group without endocrinopathy but occurred in 6 relatives of the proband with overt endocrinopathy-associated diabetes (p < 0.02) in marked association with TPO antibodies (p < 10(-4). It is concluded that relatives of probands with overt endocrine autoimmune disease-associated diabetes, unlike those of probands with diabetes alone, showed increased prevalence of thyroid antibodies and thyroid dysfunction. These results argue for a different risk of thyroid autoimmunity and clinical disease in families of diabetic patients without or with overt endocrine disease. A screening of thyroid autoimmunity is highly recommended for the latter group. PMID- 9342544 TI - Clinical aspects of diabetes secondary to idiopathic haemochromatosis in French speaking Belgium. AB - Better awareness of the clinical presentation of idiopathic haemochromatosis is a key element for early diagnosis and treatment. We retrospectively analysed the medical records of 105 patients (80 males, 25 females) diagnosed with idiopathic haemochromatosis over the past two decades in the two academic hospitals of Louvain University. Age at diagnosis was 50 +/- 12 years (mean +/- SD). Median ferritin levels were 1,803 micrograms/L-1. Cirrhosis was found at histology in 51%. Ferritin levels were significantly higher in cirrhotic than non-cirrhotic subjects (P < 0.05). Impaired glucose tolerance and diabetes were found at admission in 7 and 40% of all patients. Diabetes was more frequent when cirrhosis was present (53 vs. 25% in cirrhosis-negative patients, P < 0.05). Accordingly, cirrhosis was also more frequent in diabetic than non-diabetic patients (70 vs. 40%, P < 0.05). Diabetic subjects with cirrhosis frequently presented with symptomatic hyperglycaemia at diagnosis, had higher HbA1c levels, and were more insulin-requiring than their non-cirrhotic diabetic counterparts (P < 0.05). There was also a trend towards more frequent chronic complications of diabetes in the former group. Diabetic patients with cirrhosis had slightly higher insulin levels but lower C-peptide values (P < 0.05) than diabetic subjects without cirrhosis. Chronic phlebotomy did not affect subsequent insulin requirements. Thus, diabetes is still a frequent complication of haemochromatosis in Belgium, and its presence and severity are markedly associated with that of cirrhosis at diagnosis of idiopathic haemochromatosis. PMID- 9342545 TI - Improved metabolic control preserved beta-cell function two years after diagnosis of insulin-dependent diabetes mellitus. AB - To determine whether improved metabolic control during the first two years of insulin-dependent diabetes (IDDM) modified beta cell function, we studied 108 subjects with recent-onset IDDM diagnosed between March 1986 and April 1992 and followed up prospectively for 2 years. Two insulin regimens were used: 1) conventional insulin treatment (CIT) (1986-90, n = 67) involving a mixture of regular and intermediate insulin before breakfast and dinner; and 2) intensive insulin treatment (IIT) (1990-92, n = 41) providing regular insulin before breakfast and lunch, and a mixture of regular and long-acting insulin before dinner. Glucagon-stimulated C-peptide was determined at diagnosis and at 3, 6, 12 and 24 months. Both groups had similar clinical, metabolic and immunological characteristics at diagnosis. The IIT group had better metabolic control at any given time-point after diagnosis (mean HbA1 during follow-up in CIT: 9.86 +/- 0.28%; IIT: 8.18 +/- 0.04%; p < 0.001) (normal < 9.0%). C-peptide was increased in the IIT group 3 and 6 months after diagnosis (month 0: 0.36 +/- 0.05 nmol/l; month 6: 0.55 +/- 0.06 nmol/l; p < 0.006), but not in the CIT group (month 0: 0.39 +/- 0.04 nmol/l; month 6: 0.45 +/- 0.04 nmol/l; p = NS). Two years after diagnosis, the IIT group maintained initial C-peptide secretion (2 years: 0.37 +/ 0.04 nmol/l) whereas C-peptide was reduced in the CIT group (2 years: 0.23 +/- 0.06 nmol/l) compared to the initial value (p < 0.001) or to that of the IIT group (p = 0.017). Thus, sustained improvement in metabolic control with IIT resulted in better beta-cell function during the first two years after IDDM diagnosis. PMID- 9342546 TI - Multiple antibody status in type 1 diabetic patients and subjects at various risk with islet-cell antibodies. AB - The frequency of 37/40 kD antibodies and their association with other pancreatic humoral markers were studied in 109 recently diagnosed Type 1 diabetic patients and 116 subjects with islet-cell antibodies (ICA) at various risk for this disease (64 relatives of Type 1 diabetic patients, 23 schoolchildren with no family history of diabetes, and 29 patients with Graves' disease). At the time of diagnosis, 37/40 kD antibodies were detected in 45% of Type 1a and 77% of Type 1b diabetic patients (p = 0.03). Antibodies to glutamic acid decarboxylase (GAD) and/or 37/40 kD were present with the same frequency as ICA (86%). The frequency of 37/40 kD antibodies was not significantly different between the 3 groups at risk, in contrast with GAD antibodies which were found at a lower frequency in schoolchildren (p < 0.02). Frequencies of other pancreatic markers (ICA cross reactive with mouse pancreas and insulin autoantibodies) and the combination of ICA with at least two other markers were significantly higher in relatives than in the other groups at risk (p < 0.02). Out of 116 ICA-positive non-diabetic subjects, 10 developed diabetes. All 10 displayed 37/40kD and/or GAD antibodies during the prediabetic phase. In 8 of these 10 patients, ICA was combined with at least two other markers, whereas this association was detected in only 17 of the remaining 106 subjects who did not progress to diabetes (p < 10(-4). Thus, 37/40 kD antibodies were found in about half of Type 1 diabetic patients, and with a higher-frequency in Type 1b than 1a. In ICA-positive non-diabetic subjects, our date confirm that a combination of multiple antibodies, including GAD antibodies and 37/40 kD antibodies, can enhance the predictive value for diabetes. Comparison of ICA-positive relatives of diabetic patients, schoolchildren and patients with Graves' disease revealed distinct frequencies and combinations of markers of diabetes. This might reflect different patterns of progression among these 3 groups. PMID- 9342547 TI - Pseudohypertriglyceridaemia. AB - A mistake can be made in interpreting plasma triglyceride levels since in some cases pseudohypertriglyceridaemia may result from increased plasma glycerol due to a glycerol kinase deficit. Most automated triglyceride assays used in laboratories do not contain a negative control, i.e. a glycerol assay. We report two cases with pseudohypertriglyceridaemia due to hyperglycerolaemia and describe the clinical and biological features which suggested the diagnosis. PMID- 9342548 TI - Comment on the review by E. Cerasi, N. Kaiser and D.J. Gross (Diabetes & Metabolism, 1997, 23, 47-51) entitled "from sand rats to diabetic patients: is non-insulin-dependent diabetes mellitus a disease of the beta cell?". PMID- 9342549 TI - Metabolic investigations in humans by in vivo nuclear magnetic resonance. Recommendations of ALFEDIAM (French Language Association for the Study of Diabetes and Metabolic Diseases). AB - Although nuclear magnetic resonance (NMR) is well-established as a routine clinical imaging technique (magnetic resonance imaging or MRI), its application as a tool for metabolic investigations in humans is much less widespread in the medical community. To date, very few laboratories have combined the scientific interest, technical expertise (especially in vivo NMR spectroscopy), and facilities required to perform meaningful NMR studies of human metabolism. Among the few published reports of studies using in vivo NMR spectroscopy, diabetes mellitus, or more generally the physiology and pathophysiology of glucose homeostasis, is the dominant topic. This situation is related to the historical role of the Yale MR Center (R.G. Shulman, D.L. Rothman) in the development of in vivo NMR, and to the collaboration of NMR methodologists with the Endocrinology Section of the Yale Medical School (G.I. Shulman). Thus, in the field of diabetes and metabolism, in vivo NMR has already contributed significantly to increasing our understanding of basic physiology as well as of pathophysiological mechanisms. This "practical research note" first describes the basic principles of in vivo NMR and provides practical information about the use of this technique in metabolic investigations. The metabolic parameters accessible through in vivo NMR are then reviewed and illustrated by a few examples from the literature. PMID- 9342550 TI - The incorporation of butyrophenones and related compounds into a pharmacophore for dopamine D2 antagonists. AB - This study is an attempt to incorporate the butyrophenones, an important class of nontricyclic antipsychotic drugs, into a previously proposed pharmacophore model of tricyclic dopamine D2 receptor antagonist ligands. Conformational energy calculations were performed using the MM3-92 program on spiperone, as a representative butyrophenone, and milenperone and R48455, as related compounds with more limited conformational freedom. Twenty seven conformers were evaluated for spiperone with MM3-92 calculations and nine of these were within 1.1 kcal/mole of the global minima indicating the flexibility of the compound. A conformational analysis of twenty crystal structures of butyrophenones was also performed and six distinct conformers were represented. All of the energy minimized conformers of spiperone were superimposed in a least squares sense onto loxapine as a relatively rigid, typical D2 antagonist and a pair of mirror image conformers, which are observed in one crystal structure of spiperone, were found to be the best fit. However, it was not possible to discriminate between these two conformers since they fit the pharmacophore model equally well. The para fluoro and carbonyl group of the butyrophenones were found to correspond best to the oxygen and chlorine atoms of loxapine, respectively. The conformations of milenperone and R48455 were also consistent with the two putative biologically active forms of spiperone and the pharmacophore model. Conformational energy calculations were also performed on molindone, an antipsychotic drug in clinical use, which can be related to the butyrophenones since both have a carbonyl group adjacent to an aromatic ring. A putative biologically active form was proposed for molindone and this was related to the structure of piquindone, a rigid analog of molindone. All of the compounds were found to be entirely consistent with the pharmacophore model. However, as previously found, there is great variability in the distance between the ammonium nitrogen and the center of the relevant aromatic ring with the most extreme case in the present study being R48455 where the distance is 7.2 A. The results of the present study should also be relevant to the structures of novel, atypical antipsychotic drugs such as risperidone which appear to be analogs of the butyrophenones. PMID- 9342551 TI - Synthesis and antinociceptive activity of [D-Met2, Pro5] enkephalin [N1,5-beta-D 2,3,4,6-O-tetraacetylglycosyl]--amide and [D-Met2, Pro5] enkephalinamide. AB - Tetra-O-acetylgalactopyranosylamine and tetra-O-acetylglucopyranosylamine of D Met2, Pro5 enkephalin were designed and synthesized to enhance their membrane penetration, biological activity and resistance to proteolytic hydrolysis. Three approaches to the synthesis were attempted, which lead to a new synthetic scheme with a higher yield and enhanced ease of purification. The improved procedure involved attaching the tetra-O-acetylglycopyranosylamine to a t-Boc-Gly-Phe-Pro OH peptide, removing the t-Boc, and condensing it with t-Boc-Tyr-D-Met-OH. Biological evaluation in vivo showed that these acetylglycopyranosylamine derivatives bind to mu and delta opioid receptors in homogenate binding assays and possess analgesic activity. The analgesic potency was less than that of the parent compound D-Met2, Pro5 enkephalin. These acetylglycopyranosylamine derivatives showed enhanced lipophilicity compared to their parent compound by a partition coefficient study and they also showed greater membrane permeability, using the rabbit cornea as a model system. These derivatives also are resistant to hydrolytic enzymes as compared to the endogenous met-enkephalin when evaluated in homogenized iris-ciliary body and aqueous humor from rabbit eyes. PMID- 9342553 TI - Synthesis and QSAR of some 3-amino-2-(substituted)aminomethyl-5,6-disubstituted thieno[2,3-d]pyrimidin-4(3H)-ones as novel H1-receptor antagonists. AB - The present study describes the synthesis and quantitative structure activity relationships (QSAR) of novel 3-amino-2-(substituted)aminomethyl-5,6 disubstitutedthieno[2,3-d] pyrimidin-4(3H)-ones for their potent H1-receptor antagonist activity on the guinea pig ileum. With the IC50 values in the range of 10(-5) gms/lit, all the compounds tested were found to possess ten fold higher affinity to the H1-receptor than diphenhydramine and cetirizine, but lower than astemizole and loratidine. The sedative potential of these compounds was found to be lower than cetirizine and astemizole but comparable to loratidine. The QSAR study indicates a parabolic relationship of the biological activity mainly with the steric parameters and partly with the lipophilic parameters. PMID- 9342552 TI - Indenopyridazinone derivatives as potential antisecretory and antiulcer agents. AB - A series of substituted indenopyridazinones (4b-h) has been synthesized and tested for their antisecretory and antiulcer activity, in comparison with ranitidine, as reference drug. While the monomethoxy (4b-d), as well as the benzyloxy (4f) and the 6,9-dimethoxy (4g) derivatives were found to be devoid of overt antisecretory properties, the 9-methoxy (4e) was weakly active. The most interesting compound of this class was the 7,8-dimethoxy substituted (4h), which at an oral dose of 30 mg/kg still retains a significant activity. The dihydroderivatives of 4g,h (compounds 1g,h) were more active (1g) or comparable (1h) to the parent compounds, thus proving that the 4, 4a-double bond, which was an essential requirement in the analogues 2 and 3, is not necessary in this new series. The disubstituted derivatives (1g,h; 4g,h) were also tested as antiulcer agents, in two different models. All compounds were able to prevent haemorragic lesions induced in rats by 90% ethanol in a dose dependent manner. In the indomethacin model they still showed a significant activity, though lower than in the previous test. Attempts have been made to elucidate their mechanism of action. PMID- 9342554 TI - Ocular-specific chemical delivery systems of betaxolol for safe local treatment of glaucoma. AB - Novel ketomethoxime (BMO) and oxime (BO) analogs of betaxolol (B) were prepared through the oxidation of betaxolol, followed by quenching of the ketone with the appropriate oxyamine. The Z isomers were kinetically favored and thermodynamically more stable. Isomerization to reach an equilibrium mixture of Z/E was observed for all pure isomers in buffers. Equilibration is much faster, however in biological fluids. Ocular administration of any of the oxime derivatives, delivers betaxolol specifically to the eye tissues, with the highest concentration in the iris ciliary body. Both BMO and BO, when applied topically, showed marked reduction of intraocular pressure (IOP) in normotensive rabbits. No effect on isoproterenol-induced tachycardia in rabbits and rats were observed, even after iv. administration. Very mild eye irritation, which was less than that of betaxolol hydrochloride, was observed particularly with BMO maleate, which is an excellent candidate for safe treatment of glaucoma. PMID- 9342555 TI - Prescribing patterns for nursing home residents in the US. The reality and the vision. AB - The study of prescribing for the elderly in the US is in its infancy. Nursing home studies provide some of the most reliable data. In US nursing homes, each resident is prescribed an average of between 7.2 and 8.1 medications, with gastrointestinal agents being the most frequent. The vision for prescribing in the US involves avoidance of polypharmacy and specific groups of medications. Approaches to treatment in the US for certain conditions that can be prevented are addressed, specifically pain control, osteoporosis, stroke, cardiovascular disease, cognitive impairment and infection. PMID- 9342556 TI - Toremifene in postmenopausal breast cancer. Efficacy, safety and cost. AB - Toremifene is a chlorinated tamoxifen analogue that is indicated for the treatment of postmenopausal hormone-dependent breast cancer. It competes with estradiol for estrogen receptors and has growth-inhibitory effects on MCF-7 breast cancer cells. At concentrations < 10(-6) mol/L, this growth inhibition can be reversed by estradiol, but at higher concentrations toremifene is cytotoxic. In dimethylbenzanthracene (DMBA)-induced mammary cancer in rats, toremifene has been shown to decrease the number of new tumours and to inhibit the growth of existing tumours. Toremifene causes growth inhibition by suppressing mitosis and inducing apoptosis. The mechanism by which these events occur may involve the induction of transforming growth factor-beta 1 and inhibition of insulin-like growth factor-1 (mecasermin). Toremifene is primarily an antiestrogen, but it has some estrogen agonist properties in postmenopausal women. The latter are reflected by the fall in luteinising hormone and follicle-stimulating hormone levels and the rise in sex hormone-binding globulin levels that are associated with its use in most women. After estrogen priming, toremifene 68mg administered orally has been found to exert a similar antiestrogenic effect on the vaginal epithelium in postmenopausal women as tamoxifen 60mg. The half-life of toremifene in plasma is 5 days, and the drug is > 99% bound to plasma proteins. The main metabolites of toremifene are N-demethyl-toremifene and deaminohydroxy toremifene. Altered liver, but not kidney, function affects the pharmacokinetics of toremifene. Toremifene 60mg daily is as effective as tamoxifen 20mg daily in the treatment of postmenopausal hormone-dependent breast cancer, producing a response in about 50% of patients. Soft tissue and visceral metastases respond better to toremifene than bone metastases. Most of the adverse effects of toremifene are related to its activity at estrogen receptors and include hot flashes, vaginal discharge and nausea. Although toremifene decreases antithrombin III levels slightly, the incidence of thromboembolic complications is low. Thus far, no carcinogenic effects have been noted in humans, and preclinical data are mostly reassuring. Toremifene has favourable effects on serum lipids, and thus has potential in preventing coronary heart disease. Although toremifene is somewhat more expensive to use than tamoxifen, toremifene is an effective and well tolerated alternative to tamoxifen in the treatment of postmenopausal women with hormone-dependent breast cancer. No formal pharmacoeconomic comparisons of toremifene and tamoxifen have yet been published. Toremifene is potentially safer than tamoxifen in relation to carcinogenic effects and effects on serum lipids. PMID- 9342557 TI - Interactive wound dressings. A practical guide to their use in older patients. AB - The properties of an ideal wound dressing do not change with the introduction of new types of wound dressing, but the range of effects on wound healing increases. The number of dressings available is enormous, and the choice between them is often bewildering. Because there is still no ideal dressing for all wound types, it is necessary to get to know a few well, and to avoid switching to new therapies solely on the basis of anecdotal reports. The adoption of novel dressings should be based on scientific evidence. At present, dressings are still chosen on the basis of local traditions and personal empirical experience, together with evidence from the few double-blind, placebo-controlled trials that have been performed. In the management of ulcers, a particular wound management plan should not be changed if the ulcer being treated is decreasing in size and the patient is comfortable. The dressing should be chosen with care. The type of chronic ulcer and its appearance, the amount of exudate and the presence or absence of pain all assist in the selection of an appropriate wound dressing product. Quality-of-life aspects are important. In the elderly, good quality of life may not necessarily require complete ulcer healing, although this is naturally desirable. Dressing changes should be minimised and the ulcer should be kept moist and the surrounding skin dry. The high cost of interactive dressings is a potential disadvantage of their use. However, if the wound can be re-dressed at longer intervals and if healing occurs more quickly, their use may be cost effective and associated with less pain and a better quality of life. PMID- 9342558 TI - Current and emerging treatments for pancreatic cancer. AB - The worldwide annual pancreatic cancer death rate equals its estimated annual incidence. Surgery has been considered the only curative modality for this disease, but only 5 to 15% of patients are candidates for potentially curative resection. Evidence that postoperative adjuvant treatment improves outcome has been limited to a single randomised trial of a well tolerated split-course chemoradiation regimen. More intensive regimens have since been developed and are associated with, at best, a modest improvement in patient outcome. The potentially significant morbidity associated with pancreaticoduodenectomy, which can compromise the delivery of postoperative adjuvant chemoradiation, has led to the development of preoperative adjuvant ('neoadjuvant') chemoradiation in these patients. Although experience suggests that such an approach is feasible, its ultimate impact awaits further evaluation. Combined modality therapy has produced the most promising results in patients with unresectable or locally advanced disease. However, only modest improvements in median survival and minimal increases in long term survival have so far been achieved. This observation has encouraged many investigators to devise innovative methods of delivering therapy, including radioisotope implantation and intraoperative radiation therapy (IORT). Combined modality therapy with radioisotope implantation appears to have the greatest potential for improving local control and survival in these patients. IORT may be associated with lower morbidity than radioisotope implantation, but its impact may be limited by the radiobiological disadvantage associated with single dose boost therapy. Although new radiosensitising drugs are being tested, the problem of distant metastasis remains significant. New chemotherapeutic agents such as gemcitabine appear to have the potential to produce better results than those achieved over the last 35 years with fluorouracil. Investigations into the optimal integration of different therapeutic modalities, along with continued advances in surgery, radiation and systemic therapy, should lead to the increased use of modern multimodality interventions. In turn, this will lead us towards further improvements in outcomes for patients with pancreatic carcinoma. PMID- 9342559 TI - Hepatitis C virus infection in the elderly. Epidemiology, prophylaxis and optimal treatment. AB - Chronic infection with the hepatitis C virus (HCV) occurs throughout the world and appears to be the main cause of hepatocellular carcinoma. Studies have shown that, in areas of high endemicity, the prevalence of HCV infection is low in children but high in people aged > 60 years. Medical interventions were found to play an important role in the spread of HCV infection, because elderly patients became infected via contaminated blood transfusions or when contaminated syringes and needles were used. Maternal and sexual transmission do not appear to be the main routes of HCV infection. Interferon treatment eliminates HCV in 20 to 30% of patients with chronic HCV infection. The response to interferon therapy is usually complete in 70 to 80% of people with low levels of HCV RNA, HCV of genotype 2 and young women, but poor in elderly patients. Because liver disease can be severe in elderly patients, more effective therapies are clearly needed. PMID- 9342560 TI - Estradiol and dydrogesterone. A review of their combined use as hormone replacement therapy in postmenopausal women. AB - The focus of this review is hormone replacement therapy (HRT) with continuous oral 17 beta-estradiol (herein referred to as estradiol) 2 mg/day plus sequential oral dydrogesterone 10 or 20 mg/day for 14 days of each 28-day cycle. According to data from nonblind trials, this regimen relieves climacteric symptoms, preserves bone mineral density (BMD) and improves the cardiovascular risk profile in postmenopausal women. Increases in mean BMD in the lumbar spine of 2.4 to 6.4% have been reported after 2 years' treatment. The effect on BMD of oral estradiol plus sequential dydrogesterone was similar to that achieved with transdermal estradiol plus sequential oral dydrogesterone or with oral tibolone. Good protection against endometrial hyperplasia and cancer is provided by the dydrogesterone component. Cyclical vaginal bleeding occurs in most treatment cycles, but is generally light to moderate and the time of onset is highly predictable. Noncyclical bleeding occurs in < 10% of cycles. Mean serum high density lipoprotein-cholesterol levels are increased and low density lipoprotein cholesterol levels are decreased during treatment with oral estradiol plus sequential dydrogesterone. Insulin resistance appears to be improved. Blood pressure and bodyweight are not generally affected to any clinically important extent. Serum homocysteine levels were reported to decrease in postmenopausal women with high pretreatment levels. No data are available on the general tolerability profile of this regimen. However, the adverse events that most commonly led to discontinuation of treatment in clinical trials were typical of those associated with HRT, including vaginal bleeding headache, bloating and breast tenderness. Although the risk of breast cancer has not been specifically assessed for this regimen, it is unlikely to carry a greater risk than that of other HRT regimens. In summary available data indicate that treatment with continuous oral estradiol plus sequential dydrogesterone is effective in relieving climacteric symptoms and preserving BMD in postmenopausal women. The dydrogesterone component provides good endometrial protection and cycle control without negating the cardiovascular benefits of estradiol. Comparisons with other standard HRT regimens and long term data (including clinical end-points) are needed. In the meantime, this regimen can be regarded as an acceptable HRT option. PMID- 9342562 TI - The true yield of the small-intestinal barium study. AB - The barium examination of the small intestine is performed either as a follow through procedure or enteroclysis (small-bowel enema). The comparative diagnostic yield of these techniques is difficult to assess, mainly because of the low incidence of disorders in the small intestine. The available evidence indicates that enteroclysis is superior to the follow-through for detecting and demonstrating morphological abnormalities in the intestine. This is because the barium suspension is introduced into the intestine directly through a tube, allowing much better intestinal distension than can be achieved with the follow through. PMID- 9342561 TI - Dynamic radiology of swallowing disorders. AB - Dysphagia is a common symptom from the oral cavity, pharynx and esophagus, and its causes may be morphological or functional. A biphasic barium swallow is the best way of evaluating these patients. Using a careful clinical history, and by tailoring the examination to the individual case, the radiologist is usually able to pinpoint the cause of the patient's complaints and suggest further diagnostic procedures and treatment. PMID- 9342563 TI - Computed tomographic colonography (Virtual colonoscopy): a new method for detecting colorectal neoplasms. AB - Computed tomographic (CT) colonography is an exciting new technique that uses volumetric CT data combined with advanced imaging software to create two dimensional and three-dimensional images of the colon. The technique uses both three-dimensional images that simulate the endoluminal perspective of the colonoscope, as well as axial and reformatted two-dimensional images. The two dimensional and three-dimensional images are complementary, and in combination offer the most robust performance for the detection of colorectal polyps. Currently, CT colonographic examinations are performed in the fully cleansed and air-inflated colon using a slice thickness of 5 mm, a reconstruction interval of 3 mm, a pitch of 1.3, and 70 mA. In a blinded, prospective study of 70 patients (half with a known lesion, and half from a surveillance population with a low disease prevalence) the sensitivity for the detection of polyps of 1 cm or more is 75%, and the specificity is 90%. The most commonly encountered problems include retained colonic fluid and stool, suboptimally distended colonic segments, and long interpretation times. Many of these problems can be solved using both supine and prone imaging. It is expected that the performance of this examination will improve, and that a new era of colorectal screening will begin. PMID- 9342565 TI - Magnetic resonance cholangiopancreatography. AB - Magnetic resonance (MR) cholangiopancreatography is a new, noninvasive method of assessing the biliary tract and pancreatic duct. MRCP sequences are based on heavily T2-weighted pulse sequences, resulting in the bile ducts and pancreatic duct having very high signal intensity. Preliminary results indicate that the results of MRCP in most biliary tract diseases are similar to those of more invasive techniques of direct cholangiography, such as, endoscopic retrograde cholangiopancreatography. PMID- 9342564 TI - Dynamic rectal examination: its significant clinical value. AB - The aim of the present study was to carry out a proper correlation between patients' clinical symptoms and the radiological findings obtained by dynamic rectal examination (DRE). At DRE, the small bowel and in females the vagina are routinely opacified in addition to defecography. A prospective study of 248 consecutive patients (193 women and 55 men, ratio 3.5:1) and 14 control subjects was conducted. The parameters assessed included the anorectal angle, the position of the anorectal junction, and the total movement of the pelvic floor during squeezing and defecation. Anatomical changes as rectoceles, enteroceles and intussusceptions were also observed. Based on the findings, the following conclusions can be drawn. There is no indication for measurement of the central or posterior anorectal angle. There is no indication for measurement of the perineal ascent, perineal descent, and anorectal junction level. Anterior rectoceles occur very frequently in females, and are only of clinical relevance if the patients need digital vaginal support to facilitate defecation. DRE is a sensitive method for diagnosing enteroceles and intussusceptions. PMID- 9342566 TI - Magnetic resonance pancreatography. AB - Magnetic resonance cholangiopancreatography (MRCP) uses magnetic resonance (MR) pulse sequences in which static fluid appears bright against a low signal or dark background. When these MR images are subjected to postprocessing techniques, they produce images that resemble the pancreatograms obtained using endoscopic retrograde cholangiopancreatography (ERCP). The MRCP examination is easily performed, is noninvasive, requires no contrast injection, and has no known complications. The place of MRCP in the evaluation of disorders of the pancreas is not yet fully established, but several well-defined roles have already emerged. These include the evaluation of the pancreatic duct after failed or incomplete ERCP, and the evaluation of complete pancreatic duct obstruction. The technical aspects required for MRCP of the pancreatic duct, and the established and emerging roles for MRCP in the pancreatic duct, are described here. PMID- 9342567 TI - Magnetic resonance angiography of the visceral arteries: techniques and current applications. AB - In recent years, the techniques of contrast-enhanced magnetic resonance angiography, cine phase-contrast magnetic resonance imaging, and in-vivo magnetic resonance oximetry have been used in the diagnosis of visceral vascular diseases. The combination of these techniques can provide both anatomical and physiological information concerning the visceral circulation, and has the potential to revolutionize the work-up in patients with acute and chronic mesenteric ischemia. PMID- 9342568 TI - Hepatic magnetic resonance imaging: new techniques and contrast agents. AB - Magnetic resonance (MR) imaging is a relatively new method of examining the liver. Attempts have been made to optimize the method by developing new imaging techniques and introducing new contrast agents. The new imaging techniques have improved the image quality by shortening the examination time, reducing motion artifacts, and improving contrast-to-noise ratio. Contrast agents have improved the diagnosis of focal hepatic lesions in MR imaging in several ways. Extracellular gadolinium chelates have significantly improved the characterization of lesions, and can be optimally used as a problem-solving method for differentiating focal lesions of an unknown nature that have already been detected by other imaging modalities or by unenhanced MR imaging. Hepatobiliary and macrophage monocytic phagocytic system (MMPS)-targeted contrast agents have improved the detection of hepatic lesions. These agents are best used for preoperative evaluation of the exact number of lesions in patients with primary or secondary hepatic neoplasms. PMID- 9342569 TI - Helical computed tomography of the liver: techniques, applications and pitfalls. AB - Helical computed tomography (CT) of the liver has greatly improved both the accuracy and characterization of focal liver masses. This paper focuses on specific techniques for helical CT, including dual helical CT (acquisition of scans during both the arterial and portal venous phases of contrast enhancement), high-dose helical CT, delayed iodine CT, and helical CT angiography and portography. Techniques of generating three-dimensional helical CT angiograms from axial datasets are also discussed here. The clinical applications and results of these different techniques will also be discussed. In addition, helical CT produces or reveals some specific hepatic perfusion abnormalities that can lead to erroneous diagnoses; caveats for avoiding these pitfalls in interpretation are offered. PMID- 9342570 TI - Hepatic metastases: computed tomography versus magnetic resonance imaging in 1997. AB - The developments in computed tomography (CT) and magnetic resonance (MR) imaging that have taken place over the last two decades have dramatically increased our ability to detect and characterize focal liver lesions, and have led to the liver becoming the primary focus of interest in abdominal imaging. At the same time, advances in the medical and surgical treatment of secondary liver tumors have continued to be a challenge to these advances in radiology. It is clear that a successful outcome depends on knowledge of the size and location of the tumor burden, and accurate radiological assessment is crucial in identifying the subgroups of patients who may benefit from surgery and, at the same time, in preventing unnecessary radical surgery, with its high morbidity, in those likely to gain only a short-term benefit. The current period of limited resources, along with increased awareness of the effects of ionizing radiation, has led to competition between the two modalities, with considerable debate as to which offers the better noninvasive examination of the liver, particularly with regard to the detection and characterization of focal liver lesions. Arguments over each method's relative merits have tended to be overstated, but the parallel use of different diagnostic techniques is costly and inefficient. Each needs to be placed in an appropriate position on diagnostic pathways. PMID- 9342571 TI - Pancreatic carcinoma: applications for helical computed tomography. AB - Helical computed tomography (CT) represents a significant technical improvement over conventional (incremental) CT. This noninvasive imaging technique is now routinely used in the evaluation of patients with suspected pancreatic cancer. This paper focuses on the topics of CT technique (scanning parameters, use of intravenous contrast media, use of oral contrast media and spasmolytic agents, calculation of reformatted and projective images), as well as on the CT appearance of pancreatic carcinoma and the differential diagnosis and staging. A short comparison with other methods is included. PMID- 9342572 TI - Intraductal ultrasonography of the pancreas: development and clinical potential. AB - Intraductal ultrasound (IDUS) probes mounted with 30 MHz or 20 MHz transducers were evaluated in the diagnosis of 239 patients with pancreatic disease (including 48 cancers, 90 mucin-producing tumors, seven islet-cell tumors, two metastatic pancreatic tumors, seven serous cystadenomas, one pancreatic teratoma, three solid cystic tumors, 49 cases of chronic pancreatitis, 25 cases of focal pancreatitis, and seven cases of pancreatolithiasis). The probe was inserted via the papilla into the main pancreatic duct. In terms of resolution, IDUS at 20 MHz was able to image cystic lesions of less than 30 mm in diameter and solid lesions of less than 20 mm in diameter. With regard to vessels, IDUS was able to image the entire cross-section of the portal vein and other large veins. IDUS was useful in detecting carcinoma in situ and small tumors, in assessing the intraductal spread of the tumor and its pancreatic parenchymal invasion in mucin producing tumors of the main duct, and in assessing the indications for surgery by revealing mural nodules in mucin-producing tumors of the ductal branches. IDUS was also useful in evaluating the feasibility of partial resection of the tumor in mucin-producing tumors of the ductal branches and pancreatic islet-cell tumors, in accurately locating multiple lesions in pancreatic islet-cell cancer, and in differentiating benign from malignant cases of localized stenosis of the main pancreatic duct related to pancreatic stenting. With IDUS, the site of pancreatic stones could be identified in order to assess the need for endoscopic treatments such as stenting of the pancreatic duct and the bile duct, and the use of pulsed-dye laser treatment under pancreatoscopy for pancreatic stones. Acute pancreatitis as a complication occurred in one of the 239 patients who underwent IDUS (0.4%). An awareness of the limitations and usefulness of IDUS in evaluating pancreatic diseases can contribute to the treatment of these conditions. PMID- 9342573 TI - Interventional radiology of the biliary tract. AB - Interventional radiology provides a range of management options applicable to a broad spectrum of patients with biliary tract disorders. This review highlights the importance of these procedures, and illustrates their safety and effectiveness. Percutaneous transcatheter decompression has a well-established role in the management of patients with benign and malignant biliary obstruction. The advent of metallic stents has greatly increased the value of these techniques. Patients with biliary tract calculi can be successfully treated with a variety of percutaneous techniques, obviating surgery and providing a useful alternative to endoscopic methods. Finally, percutaneous cholecystostomy has evolved as a valuable adjunct in the treatment of calculous cholecystitis, as well as providing the definitive cure for many patients with acalculous cholecystitis. PMID- 9342575 TI - Prion diseases (transmissible spongiform encephalopathies): a review. AB - Bovine spongiform encephalopathy (BSE), or "mad cow disease", as well as Creutzfeldt-Jakob-Disease in humans, have recently been attracting public attention. The European Society of Gastrointestinal Endoscopy (ESGE) has issued guidelines on the topic (Endoscopy 1997; 29: 203-4), which were accompanied by a review article by T. Ponchon ("Transmission of hepatitis C and prion diseases through digestive endoscopy: evaluation of risk and recommended practices", Endoscopy 1997; 29: 199-202) dealing with the potential transmission of BSE and hepatitis C through gastrointestinal endoscopy. The following article on prion diseases in general provides a more in-depth overview of these disorders and their epidemiology and pathophysiology. PMID- 9342574 TI - Magnetic resonance imaging--guided abdominal interventional radiology: laser induced thermotherapy of liver metastases. AB - The aim of this study was to evaluate the clinical benefit of magnetic resonance imaging--guided laser-induced thermotherapy (LITT) for the minimally invasive treatment of liver metastases, with regard to survival rates and local tumor control. Magnetic resonance--guided LITT under local anesthesia was carried out in 134 consecutive patients aged 28-84 (mean age 69), with a total of 383 liver metastases. The major groups were liver metastases from colorectal cancer (88 patients) and liver metastases from breast cancer (20 patients), as well as metastases of miscellaneous primary tumors (26 patients). A total of 1048 laser applications were carried out. Cumulative survival times were calculated using the Kaplan-Meier method. All of the patients tolerated the procedure under local anesthesia well, and no severe complications or side effects were observed. During the follow-up period, 29 of the 134 patients treated died. The mean survival time was 35 months in the colorectal cancer group, 30 months in the breast cancer group, and 34 months in the group with miscellaneous primary tumors. The statistical assessment of the equality of survival distribution showed no significant differences between the three groups (Breslow test P = 0.35, Tarone-Ware test P = 0.49). These results suggest that in patients with liver metastases, local tumor destruction using minimally invasive percutaneous LITT under local anesthesia results in improved clinical outcomes, independently of the type of primary tumor. PMID- 9342576 TI - Attitude survey of adverse drug-reaction reporting by health care professionals across the European Union. The European Pharmacovigilance Research Group. AB - OBJECTIVES: This survey was conducted to assess the attitudes of medical practitioners in the European Union regarding their national spontaneous reporting scheme, to identify reasons for under-reporting and to determine what steps might be effective in increasing reporting rates. National spontaneous reporting schemes rely on health care professionals reporting individual cases of suspected ADRs to a central or regional agency. National schemes, however, vary considerably and reporting rates and patterns differ between member states. Accumulating evidence suggests that doctors' attitudes to national ADR reporting schemes are significant determinants of reporting rates. METHODS: A self administered questionnaire and letter of invitation was sent to a random sample of approximately 1% of medical practitioners in each of nine EU member states (Denmark, France, Ireland, Italy, the Netherlands, Portugal, Spain, Sweden and the UK). One month later, a reminder letter and a second copy of the questionnaire was sent to the non-responders (except Denmark and Italy). RESULTS: Response rates, and the percentage of responders who stated that they had reported previously an ADR, varied substantially between countries. Issues that appeared to discourage reporting included lack of availability of report forms; the address or telephone number of the reporting agency; lack of information on how to report; and not having enough time to report. Issues which did not apparently discourage reporting included concern about patient confidentiality; fear of legal liability or appearing foolish; reluctance to admit that harm had been caused to a patient; and ambition to collect and publish a personal series of cases. CONCLUSIONS: The results of this survey demonstrate some of the advantages and disadvantages of transnational, multilingual studies of this type, but indicate that there is scope for the further development of such techniques and their use on a wider basis in the EU and elsewhere. PMID- 9342577 TI - Structure and activities of hospital drug committees in Germany. AB - OBJECTIVES: Hospital drug committees have been established to ensure rational drug use. However, with regard to their structure and duties remarkable differences between European countries may exist, reflecting the differences in drug legislation and market. Our aim was to obtain information about the structure, present activities and decision-making processes of hospital drug committees in Germany and especially the role of clinical pharmacologists in these committees. METHODS: In 1995, a questionnaire with 36 items was designed and sent to all 450 hospitals in Germany with more than 400 beds. One hundred forty three returned questionnaires were evaluated. RESULTS: According to hospital size, the median value for the annual drug budget (including the cost of blood and blood-derived products) in 1993 ranged between DM 2.4 million for hospitals with less than 500 beds and DM 30.0 million for university hospitals with more than 1,000 beds. In 53.2% of drug committees, a pharmacist holds the position of chairman, followed by medical specialists (32%); (clinical) pharmacologists hold this position in only 7.7% of the general hospitals, but in almost 50% of the university hospitals. In most cases, all clinical specialities are represented in the drug committee the number of members ranging between 5 and 40 (median 12). The number of drugs included in the internal drug list, ranging between 400 in hospitals with < 500 beds and about 700 in university hospitals, strongly correlated with the number of beds and, interestingly, with the number of drug committee members. Treatment guidelines were implemented mainly for antiinfectives (87%), infusion solutions (30%), anti-emetic drugs (5-HT3-receptor antagonists, 27%) and blood and blood-derived products such as intravenous immunoglobulins (23%). However, effective control of these guidelines was only performed in about 50% of the hospitals. A drug information service was provided in most hospitals, where 95% of queries were answered by pharmacists. CONCLUSION: The results of our survey showed that German hospital drug committees vary considerably with regard to their function and control mechanisms of drug use. Most of the responders would appreciate a more intensive exchange of current problems and treatment guidelines. Although the process of pharmacotherapeutic decision making should be supported by clinical pharmacologists, experts in this field are often not involved in German hospital drug committees. PMID- 9342578 TI - How Estonian and Finnish primary care doctors rate their need for common drugs. AB - OBJECTIVE: To compare ratings of the necessity of drugs in the daily practice of experienced primary care doctors in Estonia and Finland to find out the differences and similarities in the therapeutic traditions of the two different societies. METHODS: A questionnaire was sent to all Estonian district doctors born in the 1940s and to all Finnish specialized general practitioners born in the 1940s, who then evaluated the necessity of the listed drugs on a visual analogue scale. The ratings, from 0 to 100, were entered into a computer, using a graphic tablet and a pressure sensitive pointer. RESULTS: The six most highly evaluated drugs among the Estonian respondents were digoxin, glyceryl trinitrate, aspirin, calcium-channel blockers, beta-adrenoceptor blockers and frusemide; and among the Finnish general practitioners (GPs) were penicillin, insulin, glyceryl trinitrate, beta-adrenoceptor blockers, frusemide and angiotensin-converting enzyme (ACE) inhibitors. The ratings of 15 out of 33 drugs/drug groups were very similar both in Estonia and Finland. The biggest differences between the opinions of the Estonian and Finnish doctors appeared in the ratings regarding the necessity of antacids, cimetidine, insulin, sulphonylureas, reserpine. ACE inhibitors, oral contraceptives, penicillin, metronidazole, trimethoprim, indomethacin, phenobarbital and theophylline. CONCLUSION: The revealed differences are suggested to be related to the different health care systems (different task profiles of doctors, different pharmaceutical services), different education of doctors, different availability of drugs in the past and different prices, all of which influence therapeutic traditions. PMID- 9342579 TI - Pharmacokinetics and pharmacodynamic effects of the angiotensin II antagonist valsartan at steady state in healthy, normotensive subjects. AB - OBJECTIVE: Pharmacokinetics, pharmacodynamic effects and tolerability of 200 mg valsartan, once-daily for 8 days, were investigated in 16 healthy, normotensive volunteers on a normal sodium diet. METHODS: This was a double-blind, placebo controlled, randomized crossover study. Drug concentrations in plasma and urine, angiotensin II (Ang II) concentrations in plasma, systolic (SBP) and diastolic (DBP) blood pressure, heart rate (HR) in the supine position and 3 min after passive head-up tilting, as well as safety parameters (ECG, clinical chemistry and hematology, renal water and electrolyte excretion) were measured over 24 h after the first dose (day 1) and at steady state on day 8. RESULTS: Absorption and distribution of valsartan were rapid (Cmax, 2 h; t1/2 lambda 1 < 1 h), followed by a slower terminal elimination phase (t1/2 lambda 2, 6 h) on days 1 and 8, with little accumulation in plasma (increase of 20% on day 8). Less than 10% of the dose was excreted unchanged in urine. The increase in plasma Ang II (Cmax, 6 h) was significantly enhanced at steady state. Supine SBP and DBP significantly decreased on day 8 only, by an average of -3.6 and -2.4 mmHg, respectively, versus placebo, without a concomitant increase in HR. Upon passive tilting, the increase in DBP, normally reinforced by sympathetic renin release, was slightly but significantly blunted on day 1 (-2.0 mmHg) and day 8 (-4.0 mmHg) of treatment with valsartan versus placebo. The orthostatic reflex increase in HR was slightly enhanced compared with placebo by an average of 2.8 beats min-1 on day 1 and by 2.9 beats.min-1 on day 8. Valsartan was well tolerated and had no influence on ECG, clinical laboratory parameters, and water, electrolyte and uric acid excretion. CONCLUSIONS: Pharmacokinetics of valsartan are unchanged after multiple once-daily dosing, with little (expected) accumulation in plasma. Effects of 200 mg valsartan on blood pressure in healthy subjects on a normal sodium intake are small and become more prominent after repeated dosing. Indirect evidence of AT1 blockade by valsartan is demonstrated by an increase of plasma Ang II and by a blunted DBP response to passive tilting. The decrease in blood pressure at steady state enhances the increase in plasma Ang II. Valsartan is well tolerated and is devoid of effects on water, electrolyte and uric acid excretion at 200 mg per day in healthy normotensive volunteers. PMID- 9342580 TI - Pharmacokinetic and pharmacodynamic interaction of single doses of valsartan and atenolol. AB - OBJECTIVE: Valsartan (V), a specific inhibitor of the angiotensin II receptor subtype, AT1, has been developed for treatment of hypertension. Combination therapy with a beta-adrenoceptor blocking agent might be considered in cases with insufficient efficacy of V alone. Therefore, an interaction trial was performed to evaluate the effects of co-administration of V on the pharmacokinetics of atenolol (A), and vice versa, and to monitor the pharmacodynamic response of plasma angiotensin II (ANG II) concentrations and plasma renin activity (PRA), as well as of heart rate and blood pressure, under resting and exercise conditions. METHODS: Twelve healthy, normotensive, male volunteers aged 23-46 years were treated with single doses of either 160 mg V or 100 mg A alone, or with both drugs in combination (V + A) according to a three-period crossover design. Plasma concentrations of V and A were determined using HPLC with fluorimetric and UV detection, respectively, and concentration-time profiles were established over 24 h. Plasma ANG II concentrations and PRA were monitored using specific radioimmunoassays. Heart rate and blood pressure were measured at rest and during exercise on a cycle ergometer at a workload of 2.5 W/kg-1. RESULTS: For V, mean AUC and Cmax were slightly higher when A was co-administered, the ratios of log transformed values being 1.13 and 1.22 for AUC(0-inf) and Cmax, respectively. For A, mean AUC and Cmax were slightly lower when the drug was given in combination with V. The ratios of log-transformed values in this case were 0.90 and 0.92, respectively. The sharp increase in plasma ANG II concentrations and PRA, induced by administration of V, was significantly attenuated when the drug was combined with A. In the first 12 h after drug intake, heart rate and systolic blood pressure at rest were significantly decreased when V and A were co-administered compared with treatment with V alone. V given alone did not influence heart rate or systolic blood pressure during exercise, whereas A alone and V + A led to a significant reduction in those variables. Adverse experiences reported after A and V + A could be explained by the high degree of beta-adrenoceptor blockade resulting from the administration of A. CONCLUSIONS: Co-administration of single doses of V and A does not modify the pharmacokinetics of the two drugs to a clinically relevant degree. With respect to pharmacodynamics, a single dose of A attenuates the increase in plasma ANG II and PRA in response to a single dose of V, and V has no effect on the hemodynamic response to exercise. The combined treatment with single doses of 160 mg V and 100 mg A has some additive effects on resting blood pressure in healthy, normotensive subjects. PMID- 9342581 TI - Increase or decrease of HDL-cholesterol concentrations during pravastatin treatment depending on the pre-treatment HDL cholesterol levels. AB - OBJECTIVE: The effect of pravastatin was evaluated using patient data accumulated in the data base of a hospital information system (HIS). METHODS: We selected 130 patients treated with pravastatin 10 mg per day, for a minimum period of 4 weeks. RESULTS: In the t test analysis, the reduction rates of total cholesterol (TC) and low-density lipoprotein (LDL) levels for pravastatin administration were 18%, and 27%, respectively. These values were similar to previous reports. The high density lipoprotein (HDL) level, however, did not change significantly, although previous reports have shown an elevation of HDL levels. In an attempt to explain the origin of this difference, we studied the pretreatment value dependence of the cholesterol change using regression analysis. We found that pravastatin raised the HDL level in those cases where pretreatment values were lower than 58 mg.dl-1 and reduced it for higher values. We also showed that the reductions of TC, LDL and triglyceride (TG) levels correlated positively with their pretreatment values. PMID- 9342582 TI - Use of albumin in two Spanish university hospitals. AB - OBJECTIVES: The aim of this study was to characterize the use of seralbumin, evaluating how appropriate its prescription is and what possible economic repercussions may result from inappropriate use. METHODS: We performed a prospective study that included all patients receiving albumin in two University Hospitals from October 1995 to March 1996. The reasons for albumin use were considered appropriate if they coincided with the recommendations of a panel of experts. RESULTS: During the study period, 197 patients received albumin and a total of 3208 50-ml vials (20%) were used. The internal medicine and gastroenterology services prescribed this drug the most often. The most frequent prescription motives were paracentesis in cirrhotic patients (25.9%), hypoalbuminemia (24.9%) and chronic handling of cirrhotic patients (18.6%). Only 16 prescriptions (8.1%) (corresponding to 315 vials, 9.8%) were considered appropriate. One cause of inappropriate prescribing was that colloid solutions had not previously been used in 56 (30.9%) of the 186 inappropriate prescriptions. During the study period, 74,306 ECUs were spent on inappropriate indications. CONCLUSIONS: The use of albumin in our centers is incorrect and has important economic repercussions. Some educational and informative measures must be established to change this situation. PMID- 9342583 TI - Bioequivalence, pharmacokinetic and pharmacodynamic response to combined extended release formulations of felodipine and metoprolol in healthy volunteers. AB - OBJECTIVE: The primary aim of this study was to investigate whether bioequivalence is achieved for a new fixed combination of extended-release (ER) felodipine and controlled-release (CR/ZOK) metoprolol compared with the free combination of felodipine ER metoprolol CR/ZOK. The second aim was to study whether there was an interaction in pharmacokinetics and pharmacodynamics between felodipine and metoprolol when administered as ER formulation. METHODS: Two four way cross-over studies were performed in 36 young subjects and 24 elderly subjects with frequent measurement of drug plasma concentrations, blood pressures and heart rate. The pharmacokinetic analysis included enantioselective analysis in six subjects. RESULTS: Bioequivalence between the fixed combination and the free combination was observed for the two drugs (mean difference 27%) except for a minor deviation regarding Cmax of metoprolol in the elderly. No significant interaction was shown except for a small increase (6%) of metoprolol AUC in the younger subjects. Mean plasma S-/R-enantiomer ratios were almost identical for the different treatments. Blood pressure and heart rate was significantly reduced for the fixed combination compared with felodipine ER in the younger and the elderly subjects. No significant difference regarding pharmacodynamics was detected between the fixed combination and the corresponding free combination. CONCLUSION: The fixed combination consistently provides fairly constant and effective felodipine and metoprolol concentrations after once-daily administration of one tablet. It is clinically interchangeable with the free combination of metoprolol CR/ZOK tablets and felodipine ER tablets. Finally, felodipine and metoprolol do not interact on a pharmacokinetic level when administered as the fixed combination. PMID- 9342584 TI - Metabolism of carteolol by cDNA-expressed human cytochrome P450. AB - OBJECTIVES: To determine human cytochrome P450 isoform(s) (CYPs) involved in the metabolism of carteolol, the biotransformation of the compound was investigated in vitro using ten isoforms of human cytochrome P450 expressed in human AHH-1 TK +/- cell lines. In addition, the inhibitory effects of carteolol on the activities of important CYP isoforms, namely, CYP1A2, 2C9, 2C19, 2E1, and 3A4, were examined. RESULTS: Carteolol was metabolised to 8-hydroxycarteolol by CYP 2D6 with KM and Vmax values of 183 mumoles.l-1 and 26.09 pmol.min-1.pmol-1 P450, respectively. CYP1A1, 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2E1 and 3A4 were not involved in the metabolism of the compound. CYPD6-mediated carteolol 8 hydroxylase activity was inhibited by quinidine, propranolol, nortriptyline, dextromethorphan, sparteine, bufuralol, and biperiden. Biperiden competitively inhibited the catalytic reaction with a Ki value of 0.45 mumoles.l-1. Carteolol did not affect the following catalytic reactions: CYP1A2-mediated (R)-warfarin 6 hydroxylation, CYP2C9-mediated tolbutamide methylhydroxylation, CYP2C19-mediated (S)-mephenytoin 4-hydroxylation, CYP2E1-mediated chlorzoxazone 6-hydroxylation, and CYP3A4-mediated testosterone 6 beta-hydroxylation. CONCLUSION: 8 Hydroxylation is the only cytochrome P450-catalyzed metabolic reaction of carteolol by its expressed microsomes, and CYP2D6 is the principal isoform of the enzyme involved in the catalytic reaction. Carteolol has neither stimulative nor inhibitory effects on CYP1A2, 2C9, 2C19, 2E1, and 3A4 activities. PMID- 9342585 TI - A study of the factors affecting the metabolic clearance of quinine in malaria. AB - OBJECTIVE: To assess the factors that contribute to impaired quinine clearance in acute falciparum malaria. PATIENTS: Sixteen adult Thai patients with severe or moderately severe falciparum malaria were studied, and 12 were re-studied during convalescence. METHODS: The clearance of quinine, dihydroquinine (an impurity comprising up to 10% of commercial quinine formulations), antipyrine (a measure of hepatic mixed-function oxidase activity), indocyanine green (ICG) (a measure of liver blood flow), and iothalamate (a measure of glomerular filtration rate) were measured simultaneously, and the relationship of these values to the biotransformation of quinine to the active metabolite 3-hydroxyquinine was assessed. RESULTS: During acute malaria infection, the systemic clearance of quinine, antipyrine and ICG and the biotransformation of quinine to 3 hydroxyquinine were all reduced significantly when compared with values during convalescence. Iothalamate clearance was not affected significantly and did not correlate with the clearance of any of the other compounds. The clearance of total and free quinine correlated significantly with antipyrine clearance (rs = 0.70, P = 0.005 and rs = 0.67, P = 0.013, respectively), but not with ICG clearance (rs = 0.39 and 0.43 respectively, P > 0.15). In a multiple regression model, antipyrine clearance and plasma protein binding accounted for 71% of the variance in total quinine clearance in acute malaria. The pharmacokinetic properties of dihydroquinine were generally similar to those of quinine, although dihydroquinine clearance was less affected by acute malaria. The mean ratio of quinine to 3-hydroxyquinine area under the plasma concentration-time curve (AUC) values in acute malaria was 12.03 compared with 6.92 during convalescence P = 0.01. The mean plasma protein binding of 3-hydroxyquinine was 46%, which was significantly lower than that of quinine (90.5%) or dihydroquinine (90.5%). CONCLUSION: The reduction in quinine clearance in acute malaria results predominantly from a disease-induced dysfunction in hepatic mixed-function oxidase activity (principally CYP 3A) which impairs the conversion of quinine to its major metabolite, 3-hydroxyquinine. The metabolite contributes approximately 5% of the antimalarial activity of the parent compound in malaria, but up to 10% during convalescence. PMID- 9342586 TI - Temperature dependency of the release and bioavailability of nicotine from a nicotine vapour inhaler; in vitro/in vivo correlation. AB - OBJECTIVE: To investigate the temperature dependency of the dose released and the plasma levels of nicotine from a vapour inhaler. METHODS: In an open, randomised, three-way cross-over pharmacokinetic study 18 healthy subjects inhaled nicotine for 20 min (80 inhalations) every hour for 10 h (11 administrations) at three different environmental temperatures: 20 degrees, 30 degrees and 40 degrees C. In the in vitro experiment, 5, 10, 15 and 20 l air were forced through the inhaler. With a 15 l air volume, the average amount of nicotine released was 1.44, 3.49, 4.80 and 6.99 mg at 10 degrees C, 22 degrees C, 29 degrees C and 40 degrees C, respectively. The maximum dose released at the highest temperature (40 degrees C) and the largest air volume investigated (20 l) was approximately 7.5 mg. RESULTS: In vivo peak plasma levels obtained at 30 degrees and 40 degrees C were 29.7 and 34.0 ng.ml-1, compared with 22.5 ng.ml-1 at ambient room temperature (20 degrees C). At 20 degrees C, the area under the plasma concentration-time curve (AUC) of the last dosing interval was 20.5 ng.ml-1.h. At 30 degrees C and 40 degrees C, the AUCs were 26.5 and 30.3 ng.ml-1.h, respectively. The results thus showed a mean increase of the in vivo AUC by 29% at 30 degrees C and by 48% at 40 degrees C compared with the AUC at 20 degrees C. These increases should be compared to the in vitro results, showing a mean increase of 59% and 122% respectively, at 30 degrees and 40 degrees C. The in vitro results also showed that a relatively larger fraction of the dose was released into the first 5 l of air at the higher temperatures, at 40 degrees C, about 50% of the total amount released into 20 l. CONCLUSION: It was concluded that the in vitro/in vivo discrepancy was most probably due to increased aversive effects at elevated temperatures, causing the subjects to inhale smaller puff volumes. Further, the inhaler would not produce nicotine plasma levels exceeding those achieved following cigarette smoking, even in a hot climate. PMID- 9342587 TI - Lithium intoxication in an elderly patient after combined treatment with losartan. PMID- 9342589 TI - Bioavailability of a new effervescent tablet of ibuprofen in healthy volunteers. PMID- 9342588 TI - Clinically important interaction between azathioprine (Imurel) and phenprocoumon (Marcoumar) PMID- 9342590 TI - Effects of pretreatment with clonidine, lithium and quinine on the activities of antidepressant drugs in the mouse tail suspension test. AB - The aim of the present study was the investigation of pretreatment effects with clonidine (0.06 mg/kg, intraperitoneal [i.p.]), lithium (1 mEq, i.p.) or quinine (0.5 mg/kg, i.p.) on the activities of various drugs acting on noradrenergic and/or serotonergic systems in the mouse tail suspension test. Drugs used in the present study included: the tricyclic antidepressants imipramine and dothiepin, the heterocyclic antidepressant trazodone, the 5-HT reuptake inhibitor (SSRI) fluoxetine, the atypical antidepressants mianserin and iprindole, the 5-HT1A receptor agonist ipsapirone, the 5-HT2A/2C receptor antagonist ritanserin, and the 5-HT3 receptor antagonist ondansetron. Clonidine, lithium and quinine differentially enhanced the effects of several psychotropic/drugs administered at sub-active doses. The activity of iprindole (32 mg/kg, i.p.) was not potentiated by pretreatment with clonidine, lithium or quinine. Our results suggest that lithium exerted additive effects via postsynaptic 5-HT1A receptor activation, quinine via potassium ion channel blockade of 5-HT3 receptors, while clonidine did so primarily via action at 5-HT2 receptors. PMID- 9342591 TI - Efficacy of huperzine in preventing soman-induced seizures, neuropathological changes and lethality. AB - Huperzine A (HUP) is a potent reversible inhibitor of acetylcholinesterase (AChE) that crosses the blood-brain barrier. Its ability to prevent seizures and subsequent hippocampal neuropathological changes induced by the organophosphate soman was studied in guinea pigs. Results were compared to guinea pigs treated with pyridostigmine (PYR, 0.2 mg/kg, subcutaneously). HUP pretreatment at 0.5 mg/kg, intraperitoneally, totally prevented seizures and ensured the survival of all animals for 24 h after intoxication. Hippocampal tissue was then free of any neuronal damage. Comparatively, all animals pretreated with PYR exhibited epileptic activity after soman poisoning and five of six animals died. Examination of the hippocampus of the only surviving guinea pig pretreated with PYR showed extensive neuropathological changes. Although HUP or PYR induced similar inhibitions of blood AChE activity, only HUP pretreatment led to a decrease in central AChE activity. In binding studies on guinea-pig brain homogenates, HUP had no affinity for muscarinic, alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA) and gamma-aminobutyric acid (GABA)A receptors and only a very low one for N-methyl-D-aspartate (NMDA) receptors. In conclusion, HUP, unlike PYR, protects against soman-induced convulsions and neuropathological changes in the hippocampus. This efficacy seems to be related to a protection by HUP of both peripheral and central stores of AChE. PMID- 9342592 TI - Pharmacological profile of TH-142177, a novel orally active AT1-receptor antagonist. AB - The pharmacological properties of TH-142177 (N-n-butyl-N-[2'-(1-H-tetrazole-5-yl) biphenyl-4-yl]-methyl-(N-carboxymethyl-benzylamino)-acetamide), a novel antagonist of the angiotensin II (AII) AT1 receptor, were studied in vitro and in vivo, and compared to those of losartan. In the rat isolated aorta, TH-142177 produced parallel shifts to the right of the concentration-response curves for AII-induced contractions without affecting the maximal response (pA2 = 9.07). The inhibitory potency of TH-142177 in the aorta was about three times greater than that of losartan. TH-142177 completely inhibited the specific binding of [125I]AII to AT1 receptor in rat aortic membranes (Ki = 1.6 x 10(-8) M), whereas specific [125I]AII binding to AT2 receptor in bovine cerebellum and human myocardium was not affected by concentrations of TH-142177 up to 10(-5) M. Losartan also inhibited the [125I]AII binding to rat aortic membranes (Ki = 2.2 x 10(-8) M). Following the intravenous administration to anesthetized normotensive rats, TH-142177 dose-dependently inhibited the increase in systolic blood pressure induced by an intravenous bolus injection of AII that was 1.5 times less potent than losartan. Furthermore, the oral administration of TH-142177 to conscious renal hypertensive rats exerted a dose-dependent reduction of systolic blood pressure without significantly effecting the heart rate. TH-142177 was at least three times more potent than losartan. These results demonstrate that TH 142177 is a potent and selective antagonist of AT1 receptors and by oral administration has a long-lasting antihypertensive activity. PMID- 9342593 TI - Tonic block of the Na+ current in single atrial and ventricular guinea-pig myocytes, by a new antiarrhythmic drug, Ro 22-9194. AB - Ro 22-9194 reduced the Na+ current in the atrial myocytes as well as ventricular myocytes in a tonic block fashion. Ro 22-9194 had a higher affinity to the inactivated state Na+ channels (KdI = 3.3 microM in atrial myocytes, KdI = 10.3 microM in ventricular myocytes) than to those in the rested state (KdR = 91 microM in atrial myocytes, KdR = 180 microM in ventricular myocytes), which indicated that Ro 22-9194 had a higher affinity to the Na+ channels in atrial myocytes than in ventricular myocytes. Ro 22-9194 shifted the inactivation curve in the hyperpolarized direction in both atrial and ventricular myocytes. These findings suggest that Ro 22-9194 more strongly inhibited the Na+ channel of the atrial myocytes of the diseased hearts with the depolarized membranes potentials than the Na+ channels in ventricular myocytes. PMID- 9342594 TI - Role of paraventricular and dorsomedial nuclei of the hypothalamus and central nucleus of the amygdala on muscimol-induced cardiovascular responses. AB - Gamma-aminobutyric acid (GABA) plays an important role in the central control of cardiovascular functions. Previous evidence indicates that a tonically active GABAergic system exists in forebrain structures. The purpose of this study was to examine the role of the unilateral lesion of the central nucleus of amygdala, paraventricular or dorsomedial nuclei of the hypothalamus on muscimol-induced cardiovascular responses. Electrolytic ablation of nuclei was made by a monopolar isolated electrode under a stereotaxic instrument, 3-5 days before the experiments. Effects of intracerebroventricular injections of muscimol were investigated in intact, lesioned and sham-lesioned rats. On the day of the experiments, blood pressure and heart rate recordings were carried out in male Sprague-Dawley conscious rats. Muscimol produced decreases in arterial blood pressure and heart rate. The hypotensive effect of muscimol was completely inhibited in rats with dorsomedial nucleus lesions, whereas the bradycardic effect was partially prevented. The results indicate that the dorsomedial nucleus of the hypothalamus plays an important role on muscimol-induced blood pressure and heart rate responses. PMID- 9342595 TI - In vitro calcium antagonistic and antioxidant effects of Org 13061 and its enantiomers, new potential antiatherosclerotic compounds. AB - The calcium antagonistic and antioxidant properties of a new potential antiatherosclerotic agent, Org 13061 were compared with those of its (-) and (+) enantiomers (Org 13471 and Org 13581) In vitro and with appropriate reference drugs. Org 13061 antagonized contractions induced by potassium in rabbit aortic rings with an IC50 value of 0.50 microM and reduced the maximum rate of phase 0 depolarization (Vmax) of the 'slow' calcium-mediated transmembrane action potentials in cardiac tissue (IC25 = 0.82 microM). Similarly to reference drugs, Org 13061 was more selective in reducing vascular compared to cardiac contraction. In concentrations overlapping those exerting vasorelaxant actions, Org 13061 inhibited copper ion-induced human low density lipoprotein (LDL) peroxidation (0.1-1 microM) and inhibited lipid accumulation by rat aortic smooth muscle cells in culture (1-3 microM). Higher concentrations (3 microM) modestly inhibited proliferation of these cells. The (-) enantiomer was ten times more potent than the (+) enantiomer as a vasorelaxant but was equipotent in inhibiting lipid accumulation and LDL peroxidation (eg, lag phase of conjugated dienes formation increased by 29 and 61 min and by 22 and 56 min in response to 0.3 and 1 microM (-) and (+) enantiomers, respectively). The antioxidant probucol was approximately three times more potent than Org 13061 in inhibiting lipid accumulation but was 30 times less potent in antagonizing LDL peroxidation. The classical calcium channel blocking agents were totally ineffective on lipid accumulation (1-10 microM), whereas human LDL peroxidation was slightly reduced by nifedipine (0.1-3 microM) but unaltered by diltiazem (0.1-30 microM) and verapamil (0.1-3 microM). In conclusion, the racemic Org 13061 selectively blocks voltage-operated calcium channels (VOCs) in concentrations that also exert marked antioxidant activity. The (-) enantiomer is largely responsible for calcium channel block but as antioxidants, the enantiomers are equipotent. This mixed pharmacological profile of Org 13061, not shared by known calcium channel blocking agents, may be potentially useful in the treatment of atherosclerosis. PMID- 9342596 TI - Protective effects of trimetazidine on hypoxic cardiac myocytes from the rat. AB - The electrophysiological effects of the antianginal drug trimetazidine (TMZ) were investigated in cultured rat ventricular myocytes using a substrate-free hypoxia model of ischemia. The transmembrane potentials were recorded with glass microelectrodes and the contractions were simultaneously monitored with a video motion detector. The cardiomyocytes were treated with TMZ (1-5.10(-4) M final concentration) in the bath. The untreated and the drug-treated cells were submitted either to 150 min normoxia or to 150 min hypoxia followed by 90 min reoxygenation in the absence of oxidizable substrate. In normoxic conditions, TMZ did not affect the maximal diastolic potential (MDP) but significantly lowered the plateau potential level (OS) and decreased the upstroke velocity (Vmax) and the spontaneous action potential rate (APR). Conversely, TMZ significantly increased action potential duration at 80% repolarization (APD80). Under substrate-free hypoxia, the untreated cells displayed a progressive contractile failure and an important decrease in OS and APD. In parallel, early postdepolarizations triggering high rate spikes were observed. Prolonging oxygen depletion led to the cessation of the spontaneous electrical activity and thereafter to a gradual decrease in MDP. Near normal rhythmic action potentials and contractions resumed after reoxygenation. Comparatively, the treatment by 5.10(-4) M TMZ almost completely prevented the decrease in plateau amplitude, resting membrane potential, Vmax, APD80, and rate caused by substrate-free hypoxia. Moreover, the hypoxia-induced arrhythmias and the cessation of spontaneous electromechanical activities did not occur in the presence of TMZ (5.10(-4) M). After reoxygenation, the TMZ-treated cells exhibited a higher action potential amplitude than that of the untreated cells, although the TMZ induced depressive effects on the spontaneous frequency and the Vmax persisted. In conclusion, this study shows that TMZ (5.10(-4) M) is efficient in protecting the isolated cardiac myocytes against the functional alterations induced by substrate-free hypoxia and led thus to a better recovery upon reoxygenation. The cytoprotective action may be linked, at least in part, to apparent ion channel blocking effects of the drug, which appeared in basal conditions at concentrations used in this study. PMID- 9342597 TI - Comparison of the effects of cyclosporine A and trimetazidine on Ca(2+)-dependent mitochondrial swelling. AB - Cyclosporine A (CsA) is a known potent inhibitor of pro-oxidant-induced mitochondrial swelling. In the present study we show that CsA's effect is only transient when the liver mitochondrial swelling in induced by Ca2+ plus tert butylhydroperoxide (t-BH). After an initial inhibition, swelling is worsened by CsA as evidenced by an extent of mitochondrial swelling that exceeds that of the control. Unlike CsA, trimetazidine (TMZ), an anti-ischemic drug decreases both the extent and the rate of the swelling with an IC50 value of 214 +/- 24 microM. Its inhibition effect on the initial swelling rate mimicks that of CsA but the mechanism may be independent. During long-term swelling. TMZ counteracts the worsening effect of CsA. The inhibition of swelling induced by TMZ is assessed by the fact that TMZ significantly increases the EC50 of Ca(2+)-induced mitochondrial swelling (46.6 +/- 6.0 to 85 +/- 10 microM, P < 0.01), without affecting its cooperativity. Apparently, TMZ seems to behave like trifluoperazine (TFP), a phospholipase A2 inhibitor that, under our experimental conditions, inhibits the mitochondrial swelling induced by Ca2+ and t-BH with an IC50 value of 25 +/- 10 microM. Both drugs are able to protect mitochondria from both phases (early and late) of the swelling, especially the late, which is enhanced in the presence of CsA. TFP and other phospholipase A2 inhibitors were able to displace [3H]TMZ from its mitochondrial binding sites whereas CsA was ineffective. We suggest that TMZ, like TFP, inhibits the CsA insensitive mechanism involved in the swelling process which is responsible for the worsening effect observed in the presence of CsA when the swelling is generated by Ca2+ and t-BH. PMID- 9342598 TI - Synergistic actions of pentobarbital and dihydropyridine Ca2+ antagonists on guinea pig isolated thoracic aorta. AB - In order to elucidate the mechanism(s) behind the interactions between barbiturates and Ca2+ antagonists, the effects of pentobarbital combined with three structurally diverse types of Ca2+ antagonist on CaCl2-induced contractile responses of the guinea pig thoracic aorta in Ca(2+)-free and 40 mM K+ medium and the effects of pentobarbital on Ca2+ antagonist binding to guinea pig aortic membranes were investigated. The dihydropyridine derivatives isradipine (10(-10) 10(-8) M) and nifedipine (10(-10)-10(-8) M) inhibited CaCl2-induced contractions concentration-dependently. Treatment with both pentobarbital (10(-4) M) and dihydropyridine Ca2+ antagonists (10(-9) M) shifted the CaCl2 concentration response curves to the right significantly compared with those after treatment with the Ca2+ antagonists and pentobarbital alone. However, no synergistic effects of pentobarbital (10(-4) M) with other types of Ca2+ antagonist (verapamil (10(-7) M) and diltiazem (10(-6) M)) were observed. The binding of [3H]isradipine (2 x 10(-9) M) to guinea pig aortic membranes was increased significantly by simultaneous pentobarbital treatment, but no such effect was observed with [3H]verapamil (10(-8) M) or [3H]diltiazem (2 x 10(-8) M). These findings suggest that the synergistic contractile effects of pentobarbital and dihydropyridines were, in part, due to enhancement of dihydropyridine binding to guinea pig aortic membranes (L-type Ca2+ channels) by pentobarbital and that the interactions between pentobarbital and Ca2+ antagonists may be structurally specific. PMID- 9342599 TI - 2-Methyl-thiazolidine-2,4-dicarboxylic acid as prodrug of L-cysteine. Protection against paracetamol hepatotoxicity in mice. AB - Toxic doses of paracetamol (acetaminophen) destroy the cellular defense system in hepatic tissue. The degree of the destruction can be assessed be measuring the metabolism of sulfhydryl compounds, oxygen radicals and the release of certain enzymes. Administration of 2-methyl-thiazolidine-2,4-dicarboxylic acid (CP; 1.2 mmol/kg) to mice 12 h prior to a toxic dose of paracetamol (600 mg/kg) suppressed the increase of aminotransferase activities in blood serum and the levels of reactive oxygen species in liver tissue. A protective effect of CP was also observed with respect to depletion of non-protein sulfhydryl compounds, cysteine and glycogen. The findings demonstrate that the cysteine prodrug CP is effective in preventing liver damage of a hepatotoxic dose of paracetamol in vivo. A further advantage of the new compound is the long duration of the effect of more than 12 h. PMID- 9342600 TI - Trolox-derivative antioxidant protects against methanol-induced damage. AB - This paper reports data on the effect of a new antioxidant, U-83836E, on the lipid peroxidation and antioxidant status of liver, red blood cells (RBCs) and blood serum of rats intoxicated with methanol (3.0 g/kg body weight). Methanol administration slightly increased the levels of peroxidation products in the liver, and markedly increased them in RBCs and serum. In contrast, glutathione peroxidase, glutathione-reductase activity, reduced glutathione concentration and total antioxidant status were decreased. The use of U-83836E, containing a trolox ring, appeared to be beneficial in reducing lipid peroxidation products and in partially in preventing the decrease in glutathione and antioxidant enzymes induced by methanol in liver and serum. These results show that antioxidant U 83836E may partially prevent methanol toxicity. PMID- 9342601 TI - Effects of lorazepam on film-induced differentiated emotions in healthy volunteers. AB - We studied the effects of lorazepam, a benzodiazepine, on differentiated emotions in healthy volunteers. In order to induce differentiated emotions, film excerpts were selected on the basis of the type of emotion they induced (fear, anger and for affective tone neutral film). For 6 days (D1 to D6), ten healthy volunteers received lorazepam (1 mg bid) or placebo in a randomized cross-over double-blind trial. During each treatment period, emotional induction occurred on D4, D5 and D6. One film excerpt (fear, anger or neutral) was presented each morning after relaxation. Evaluation was performed before and after each emotional induction and included questionnaires (Differential Emotions Scale and physical activation visual analog scales), and neurophysiological parameters (systolic and diastolic blood pressure, heart rate and norepinephrine levels). Globally, the film excerpts induced the predicted emotions. An analysis of variance was undertaken and revealed a significant effect of lorazepam versus placebo. On the Differential Emotions Scale and during fear induction, lorazepam induced a significantly higher increase in fear, anxiety and disgust emotions than placebo, whereas no effect was observed after anger induction. Lorazepam also induced a significantly higher increase in diastolic and systolic blood pressure with no change in heart rate, and physical activation items ("tears" and "faster breathing") without no significant change in norepinephrine. In conclusion, our results are consistent with an overall increase in emotional reactivity with lorazepam (1 mg bid) as compared to placebo. The pertinence of film-induced differentiated emotions has to be confirmed for clinical pharmacological use. PMID- 9342602 TI - Molecular genetics of craniosynostotic syndromes. AB - This article reviews recent molecular genetic findings in autosomal dominant craniosynostotic syndromes. A mutation in the homeotic gene MSX2 was the first genetic defect identified in an autosomal dominant primary craniosynostosis, i.e. in craniosynostosis type 2 (Boston type). In the more common syndromes of Crouzon, Pfeiffer, Jackson-Weiss, and Apert, mutations were found in the gene coding for fibroblast growth factor receptor (FGFR) 2. Less frequently, mutations are observed in FGFR1 and FGFR3 in some cases of Crouzon and Pfeiffer syndrome. The mutations identified in FGFR2 are located in exons 5 and 7 of the gene that code for immunoglobulin (Ig)-like chain III and the region linking Ig II and Ig III of the receptor. These domains of the receptor are important for ligand binding. Apart from Apert syndrome, identical mutations are found in the clinically distinct syndromes of Crouzon, Pfeiffer, and Jackson-Weiss. Furthermore, the same gene defect can result in a highly variable phenotype even within one family. Therefore, the clinically distinct craniosynostotic syndromes are extremes of a spectrum of craniofacial abnormalities and not nosologic entities. In Saethre-Chotzen syndrome, the gene coding for transcription factor TWIST is mutated. The disease genes identified in craniosynostotic syndromes to date either regulate transcription or are required for signal transduction and play a central role in the development of the calvarial sutures. PMID- 9342603 TI - Endothelin-like immunoreactivity in aqueous humor of patients with primary open angle glaucoma and cataract. AB - BACKGROUND: Experimental evidence suggests a role of endothelin-1 (ET) in the regulation of intraocular pressure (IOP). METHOD: Therefore, in patients undergoing cataract surgery, ET-like immunoreactivity (ETIR) was measured by radioimmunoassay in pooled samples of aqueous humor of eyes with primary open angle glaucoma (POAG) and normotensive eyes with cataract only. RESULTS: ETIR was significantly (P < 0.05) higher in patients with cataract and POAG (20.5 +/- 1.8 pg/ml, n = 12; preoperative IOP 21.4 +/- 1.1 mmHg, n = 33) than in patients with cataract only (15.8 +/- 1.6 pg/ml, n = 15; preoperative IOP 16.0 +/- 0.6 mmHg, n = 77). CONCLUSION: This finding may indicate a role of ET in POAG or ocular antihypertensive treatment, and its relevance should be further investigated. PMID- 9342604 TI - The influence of glaucoma history on graft survival after penetrating keratoplasty. AB - BACKGROUND: It was the purpose of this retrospective study to evaluate the effect of a preoperative history of glaucoma on graft survival after penetrating keratoplasty. PATIENTS AND METHODS: Six hundred and forty-six penetrating keratoplasties with generally good prognosis were analyzed retrospectively. Indications for surgery were corneal dystrophy, degeneration and scarring. Only first keratoplasties in corneas without severe vascularization or acute inflammation were included. Surface disorders, a history of herpes or Acanthamoeba keratitis were further exclusion criteria. Keratoplasties were performed only if glaucoma seemed to be controlled preoperatively. Graft survival ratios were calculated according to Kaplan and Meier, and statistical significance was evaluated by means of the log-rank test. RESULTS: With a glaucoma history the estimated 3-year graft survival rate was 71%, in contrast to 89% without such a history. This difference was statistically significant (P < 0.001). There was no difference between the groups with respect to immune reactions. With a glaucoma history, postoperative episodes of glaucoma decompensation were responsible for half of the graft failures. CONCLUSIONS: A preoperative history of glaucoma affects graft prognosis negatively, presumably through a negative influence of postoperatively elevated intraocular pressure on a vulnerable graft endothelium, and not by an increase in immune reactions. Therefore, keratoplasties in eyes with glaucoma are high-risk procedures and glaucoma has to be monitored more efficiently pre- and postoperatively. PMID- 9342605 TI - Iris pigment epithelium transplantation. AB - BACKGROUND: Iris pigment epithelium (IPE) cells and retinal pigment epithelium (RPE) cells possess the same embryonic origin. It is also known that the pigmented epithelial cells in the eye have a high transdifferentiation potential. In this study we transplanted IPE cells into the subretinal space of albino Royal College of Surgeons (RCS) rats and evaluated their influence on the degeneration of the photoreceptors. METHODS: IPE cells of Long Evans rats were isolated and pure cultures were obtained. The isolated cells were transplanted into the subretinal space of RCS rats. Light microscopic and morphometric analysis were carried out. RESULTS: The IPE transplants survived in the subretinal space and attached themselves to the Bruch's membrane. The transplanted cells were able to delay the degeneration of the photoreceptors for up to 3 months. CONCLUSION: These results suggest that IPE cells could be successfully transplanted and survive in the subretinal space. In the transplanted eyes the photoreceptors were preserved for a period of 3 months. Further studies are needed to explore the capability of IPE cells to assume the main functions of RPE cells in the subretinal space and their potential in the therapy of selective degenerative diseases of the retina. PMID- 9342606 TI - How often do patients need visual field tests? AB - BACKGROUND: This study was undertaken to determine whether the interval between visual field tests affects the ability to detect progressive glaucomatous field loss. METHODS: One hundred and nineteen retinal locations which were deteriorating significantly by > or = 1 dB/ year (untreated normal tension glaucoma patients: 6 eyes) were studied. Analysis was repeated using 'thinned' visual field tests: one test per year instead of the complete three per year over a period of 4 years. RESULTS: The 'thinned' tests identified only 45.4% of the deteriorating points over the 4-year period. Furthermore, there was a mean delay of 1.10 years in detection (P < 0.01). CONCLUSIONS: Less frequent visual field testing detects fewer progressing locations and detects them later. PMID- 9342607 TI - Vitrectomy for traction macular detachment in diabetic retinopathy. AB - BACKGROUND: A small number of eyes with proliferative diabetic retinopathy develop massive central fibrovascular membranes characterized by vitreoretinal tractions along the arcades and optic disk and retinal traction lines extending through the macula. The aim of our study was first to present the results of vitrectomy for removal of these central membranes and second to determine the correlation between preoperative parameters and postoperative visual outcome. SUBJECTS AND METHODS: We treated 28 eyes with severe central fibrovascular diabetic membranes by a modified bi-manual en bloc excision technique during vitrectomy. Preoperative examination included general status, visual acuity, slit lamp investigation, binocular funduscopy, ultrasound investigation and visual evoked potentials (VEP). Further, we analyzed intraoperative complications and postoperative anatomic and functional outcomes. RESULTS: The retinas of 27 eyes with central traction retinal detachments were reattached by surgery. With a minimum of 6 months' follow-up, the macula remained attached in 24 eyes, while the retinas were completely attached in 22 eyes. Preoperative visual acuity was defective light perception to 0.1; an increase in visual acuity to maximal 0.1 was seen in 50% of the patients postoperatively. Preoperative visual acuity of light perception was associated with no functional improvement. Preoperative ultrasound investigation gave information about the real anatomic situation of the retina, especially if funduscopy was not possible. The other preoperative parameters could not predict correctly the functional outcome of vitrectomy in diabetics with severe central fibrovascular membranes because of the damage of the optic nerve and the retina. CONCLUSIONS: The high rate of anatomical reattachment after vitrectomy in diabetic eyes with severe central fibrovascular membranes is associated with a slight improvement of function; only preoperative visual acuity of hand motions or better was associated with an improvement of function. PMID- 9342608 TI - Ocular findings in patients with autosomal dominant retinitis pigmentosa and Cys110Phe, Arg135Gly, and Gln344stop mutations of rhodopsin. AB - This report describes ocular findings obtained in four patients from three families with autosomal dominant retinitis pigmentosa (adRP) due to missense mutations in the rhodopsin gene. Phenotypes were characterized by standard ophthalmologic examinations, visual fields, electroretinography (ERG), dark adaptation, and two-color dark-adapted threshold perimetry. Two patients aged 38 and 45 years, respectively, from a family with the Cys110Phe mutation showed mild fundus changes without bone spicules as well as small arcuate scotomas in the inferior quadrants of their visual fields but displayed severe functional loss of rods and cones in the ERG. Two-color dark-adapted threshold perimetry revealed a regional type of degeneration. A 48-year-old patient with an Arg135Gly mutation had typical RP with concentrically narrowed visual fields and nondetectable ERG responses. Central visual functions were well preserved for a long time. Two color dark-adapted threshold perimetry indicated a diffuse type of retinal degeneration. An 18-year-old patient with a Gln344stop mutation has been followed for 13 years. His ERG was clearly reduced at the age of 5 years; since that time, disease progression has been very slow. Currently, there are relatively mild alterations in visual acuity, rod sensitivity, and visual fields. Our findings confirm that there is a large phenotypic variety among patients with adRP and different rhodopsin mutations. PMID- 9342609 TI - Inhibitory effects of plasminogen fragment on experimentally induced neovascularization of rat corneas. AB - BACKGROUND: Corneal neovascularization plays an important role in the pathogenesis of a number of corneal disorders. Recently a polypeptide was demonstrated, generated by the primary tumor, that inhibited angiogenesis and growth in metastases. This polypeptide is similar to a 38-kDa plasminogen fragment. METHODS: We surgically implanted into rat corneal stroma a slow-release ethylene-vinyl-acetate (EVA) copolymer pellet containing basic fibroblast growth factor (bFGF) to induce corneal neovascularization. Then we applied aqueous solution containing plasminogen fragment to the rat cornea in order to observe the degree of inhibition of angiogenesis. RESULTS: In the eyes of control rats, neovascularization from the limbus to the pellet occurred, graded 4+ in all five animals. In plasminogen fragment-treated rats, there was virtually complete inhibition of the neovascular response to the pellet. Of five treated rats, three showed no neovascularization and two demonstrated grade 1+ neovascularization. The difference in the degree of neovascularization between control and plasminogen fragment treatment was statistically significant (P < 0.05). CONCLUSION: Our studies provide the first direct evidence that rat corneal neovascularization is inhibited by instillation of plasminogen fragment. This agent may prove useful in the treatment of corneal angiogenic disorder. PMID- 9342610 TI - Retinal pigment epithelial cells: autocrine and paracrine stimulation of extracellular matrix contraction. AB - BACKGROUND: This study was carried out to examine the biological activity of contraction promoters produced by dedifferentiating retinal pigment epithelial cells (RPE) and to evaluate the importance of autocrine and paracrine effects within a semi-closed environment like the vitreal cavity. METHODS: RPE at different stages of dedifferentiation in culture were examined for their ability (a) to generate tractional forces in vitro, with and without serum stimulation, and (b) to produce and release contraction-stimulating proteins. Autocrine versus paracrine effects of cell-secreted promoters were tested by using RPE or human dermal fibroblasts (HDF) as target cells. The contraction-stimulating activity of the cell-secreted promoters was partially characterized and compared to the activity of defined promoters. RESULTS: Our study confirmed that RPE can synthesize and secrete cell-contraction-promoting factor(s) active in stimulating the development of tractional forces by RPE as well as HDF. The quantity of biological activity secreted per cell decreases with progressive dedifferentiation, yet the responsiveness of the cell to contraction promoters increases. The contraction promoter(s) synthesized by RPE is partially distinct from the promoters in serum, TGF-beta 1 and beta 2, IGF-1, ET-1 and PDGF. The contraction-promoting effects of the RPE product(s) can be completely blocked by staurosporine. CONCLUSION: De-differentiation of RPE is characterized by increasing capacity to generate tractional forces and decreasing synthetic capacity. RPE within a semi-closed system like the vitreal cavity can, theoretically, act both as promoting and active component of traction-related events (tractional retinal detachment). PMID- 9342612 TI - Retinal neovascularization in a case of macular branch retinal vein occlusion. AB - BACKGROUND: Retinal neovascularization (RNV) has never been described in cases of macular branch retinal vein occlusion (MBRVO), due to the limited amount of ischemia in this form of retinal vein occlusion. Ischemic areas as wide as 5-10 disc diameters were required by previous studies to count as ischemic cases of central or major branch retinal vein occlusion. CASE REPORT: A 56-year-old woman who had been suffering from MBRVO for 3 years presented at the posterior pole a zone of non-perfusion, extending over 7.5 disc areas, and three small tufts of RNV. RNV regressed after two subsequent laser treatments of the ischemia. Retrohyaloid hemorrhage was observed 2 months after the first treatment. CONCLUSION: Since the average diameter of the non-perfused area was about 2.75 disc diameters, this case demonstrates that small RNVs can appear in less extensive areas of ischemia than is generally believed. PMID- 9342611 TI - Early experience with intravenous immunoglobulin treatment in Wegener's granulomatosis with ocular involvement. AB - BACKGROUND: Pooled intravenous gammaglobulin (IVIg) was reported to be effective in the treatment of Wegener's granulomatosis (WG). No reports have been made on the effects of this new treatment on ocular manifestations of WG. METHOD: IVIg treatment was given to two patients suffering from WG with ocular involvement after several other treatment regimes had failed. RESULTS: Although the systemic disease was under control, the ocular symptoms of both patients worsened during and after IVIg treatment. In one case an adverse effect consisting of retinal vasculitis was noted on two occasions. CONCLUSION: Although beneficial effects of IVIg treatment on WG have been previously described, the two cases with ocular involvement presented here did not reveal any positive response. Paradoxical and unpredictable reactions cannot be ruled out. Thus, patients treated with IVIg should be closely surveyed by an ophthalmologist. PMID- 9342613 TI - Diagnosis of MALT lymphoma by conjunctival biopsy: a case report. AB - BACKGROUND: Most extranodular lymphatic tissue is found in the intestinal mucosa. Together with similarly structured lymphatic tissue at other locations it has been named mucosa-associated lymphatic tissue (MALT). Malignant transformation of such tissue to lymphoma is well known. Although MALT lymphoma has been described in tissue physiologically void of MALT, lymphoma manifestation in the conjunctiva is rare. METHODS: We report a case of a 47-year-old woman who was referred to our clinic for symptomatic treatment and evaluation of severe symptoms of dry eyes. She was thought to suffer from Sjogren's syndrome because of xerophthalmia and xerostomia, as well as massive bilateral swelling of the parotid gland. Ophthalmological examination revealed marked hyperplasia of the conjunctiva, of which a biopsy was taken. RESULTS: Histological and immunohistochemical examination of the conjunctival biopsy, together with analysis of gene rearrangement by Southern blot, led to the diagnosis of low-grade B-cell lymphoma of the MALT. CONCLUSION: The differential diagnosis of keratoconjunctivitis sicca presenting with conjunctival swelling of unknown origin should include lymphoma, especially since Sjogren's syndrome may be associated with malignant disorders of the lymphatic system. A biopsy of suspicious conjunctival changes can clarify a multisystem disease by providing a tissue diagnosis. PMID- 9342615 TI - Robert Feulgen Lecture 1997. Lipid microdomains and membrane trafficking in mammalian cells. AB - This overview summarizes the data for how epithelial cells sort and deliver proteins and lipids to the apical and basolateral cell surface domains. The basolateral pathway uses a Rab-SNARE mechanism for docking and fusion, while the apical route employs a different machinery. This latter mechanism is based on lipid microdomains, composed of clusters of sphingolipids and cholesterol, which function as rafts for apical delivery. The sphingolipid-cholesterol raft mechanism seems to be employed generally by mammalian cells to transport raft associated proteins to their post-Golgi destinations. PMID- 9342614 TI - Application of in situ hybridization, cytochemical and immunocytochemical techniques for the investigation of peroxisomes. A review including novel data. Robert Feulgen Prize Lecture 1997. AB - In situ hybridization, cytochemical and immunocytochemical techniques have contributed significantly to the understanding of the biology of peroxisomes, since they permit in situ demonstration of the sites of synthesis and distribution of peroxisomal proteins without the necessity of homogenization and subcellular fractionation of tissues or cultured cells. This article reviews the results of research on mammalian peroxisomal metabolism, biogenesis and proliferation in which morphological techniques have played a significant role in the elucidation of the biological problem. Some new data on peroxisomal heterogeneity and morphogenesis are included. The morphological methods applied have made it possible to characterize the differences in distribution of mRNAs encoding peroxisomal proteins in different tissues, as well as to monitor the marked heterogeneity in the protein composition and in the activity of specific enzymes in the peroxisomal population of single cells, or in tissues with complex organization (e.g. liver and kidney). In addition, the dynamic alterations and high plasticity of the peroxisomal compartment--partly dependent on contact of the peroxisomes to the microtubular network-are presented. PMID- 9342616 TI - Cofilin undergoes rapid dephosphorylation in stimulated neutrophils and translocates to ruffled membranes enriched in products of the NADPH oxidase complex. Evidence for a novel cycle of phosphorylation and dephosphorylation. AB - Neutrophils contain a 21-kDa phosphoprotein that undergoes rapid dephosphorylation upon stimulation of these cells with the chemoattractant N-fMet Leu-Phe (fMLP), activators of protein kinase C [e.g., 4 beta-phorbol 12-myristate 13-acetate (PMA)] or the calcium ionophore A23187. This phosphoprotein was identified as the non-muscle form of cofilin by peptide sequencing and immunoblotting with specific antibodies. Evidence is presented that in neutrophils cofilin is regulated by a continual cycle of phosphorylation and dephosphorylation, and that the phosphatase undergoes activation during cell stimulation. Experiments with a wide variety of antagonists further suggested that the protein kinase that participates in these reactions may be a novel enzyme. The kinetics of cofilin dephosphorylation in neutrophils stimulated with fMLP or PMA were very similar to those observed for superoxide (O2-) release. Immunofluorescent studies revealed that cofilin was present throughout the cytosol of resting neutrophils and underwent rapid translocation to the F-actin rich, ruffled membranes of stimulated cells. Cytochemical analysis further revealed that the ruffled membranes also contained large amounts of hydrogen peroxide (H2O2), a product of the O2-/H2O2-generating activity of stimulated neutrophils (NADPH oxidase). Cofilin is therefore well placed to participate in the continual polymerization and depolymerization of F-actin that is thought to give rise to the oscillatory pattern of H2O2 production observed under certain conditions. PMID- 9342617 TI - Characterisation of cellular infiltration and adhesion molecule expression in CBA/CaH-kdkd mice with tubulointerstitial renal disease. AB - CBA/CaH-kdkd mice develop a spontaneous and chronic tubulointerstitial renal disease which is characterised by mononuclear cell infiltration, tubular collapse and cystic dilatation of tubules. The pathogenic mechanisms of renal injury have not been fully elucidated in this model. We have analysed the nature of infiltrating cells and the expression of MHC class II antigens, cytokines and adhesion molecules in CBA/CaH-kdkd kidneys at various disease stages. Using immunohistochemical techniques we found that kdkd kidneys are characterised by abundant macrophage and dendritic cell infiltration with fewer T cells with CD4+ and CD8+ phenotypes. Interestingly, MHC class II antigens were not induced on renal tubules. The proinflammatory cytokine, TNF-alpha, was markedly enhanced in kdkd kidney (up to fourfold), whereas the T cell-specific cytokine, IFN-gamma, increased less (less than twofold). ICAM-1 and VCAM-1 were markedly overexpressed by injured proximal tubules. ICAM-2 and PECAM-1 were constitutively expressed on glomerular capillaries and vascular endothelium in normal kidneys and did not change in CBA/CaH-kdkd mice. In conclusion, tubulointerstitial nephritis in CBA/CaH-kdkd mice is characterised by prominent macrophage infiltration and abundant expression of ICAM-1 and VCAM-1 on injured renal tubules. The lack of MHC class II antigens on injured tubules suggests that the kd gene defect could generate a secondary renal inflammatory response which is characterised by prominent macrophage infiltration and a relative scarcity of T cells. PMID- 9342618 TI - Dystrophin in rod spherules; submembranous dense regions facing bipolar cell processes. AB - It is known that the retina contains the protein dystrophin in the ribbon synapse, but the ultrastructural analysis is not yet fully elucidated. Our previous study reported that dystrophin is localized under the rod cell membranes in rat retinas. In the present study, we have investigated the relationship between dystrophin-rich regions of rod cell membranes and other neuronal processes in mouse retinas with a monoclonal antibody raised against the human dystrophin C-terminus. Immunoblotting, immunofluorescence stainings, and immunoelectron microscopy were employed. Immunoblotting analysis indicated that mouse retinas possessed some of the dystrophin isoforms of approximately 260 kDa, 140 kDa, and 70 kDa molecular weight. Confocal images showed a punctate appearance in the outer plexiform layer, as previously described. Immunoelectron microscopy showed that dystrophin immunoreactive products were always observed at submembranous dense regions of the rod spherule abutting bipolar processes. These results suggest that retinal dystrophin may be closely involved in signal transmission from rods to bipolar cells. PMID- 9342619 TI - Localization of dystrophin and beta-dystroglycan in bovine retinal photoreceptor processes extending into the postsynaptic dendritic complex. AB - Dystrophin is an actin-binding protein of the membrane cytoskeleton that binds to dystroglycan, an integral membrane protein of the plasma membrane that is posttranslationally cleaved into a transmembrane dystrophin-binding beta-moiety and an extracellular laminin- and agrin-binding alpha-moiety. Mutations of dystrophin may not only cause Duchenne muscular dystrophy but may also be associated with abnormal electroretinograms assumed to result from disturbed neurotransmission between retinal photoreceptors and bipolar cells. Here we show by confocal laser microscopy and immunogold electron microscopy that dystrophin and beta-dystroglycan are colocalized in bovine rod photoreceptor synaptic complexes distal from the ribbon-containing active synaptic zones. Both proteins are restricted to a microdomain of the photoreceptor plasma membrane that forms the lateral wall of the synaptic cavity and projects with finger-like extensions into the postsynaptic dendritic complex. Within the cavity these processes eventually come into close contact with bipolar cell dendritic endings. We speculate that the dystrophin-dystroglycan complex of the cavital plasma membrane stabilizes the elaborate synaptic morphology or plays a role in the immobilization of still unknown transporters and receptors involved in certain aspects of neurotransmission to bipolar cells. A further outcome of this study is that dystrophin and dystroglycan are located along the vitread membrane surface of Muller cell endfeet where this protein complex may be important for the attachment of the retina to the basal lamina and the vitreous. PMID- 9342620 TI - Occupational health in Brazil. AB - Brazil is a recently industrialised country with marked contrasts in social and economic development. The availability of public/private services in its different regions also varies. Health indicators follow these trends. Occupational health is a vast new field, as in other developing countries. Occupational medicine is a required subject in graduation courses for physicians. Specialisation courses for university graduated professionals have more than 700 hours of lectures and train occupational health physicians, safety engineers and nursing staff. At the technical level, there are courses with up to 1300 hours for the training of safety inspectors. Until 1986 about 19,000 occupational health physicians, 18,000 safety engineers and 51,000 safety inspectors had been officially registered. Although in its infancy, postgraduation has attracted professionals at university level, through residence programmes as well as masters and doctors degrees, whereby at least a hundred good-quality research studies have been produced so far. Occupational health activities are controlled by law. Undertakings with higher risks and larger number of employees are required to hire specialised technical staff. In 1995 the Ministry of Labour demanded programmes of medical control of occupational health (PCMSO) for every worker as well as a programme of prevention of environmental hazards (PPRA). This was considered as a positive measure for the improvement of working conditions and health at work. Physicians specialising in occupational medicine are the professionals more often hired by the enterprises. Reference centres (CRSTs) for workers' health are connected to the State or City Health Secretariat primary health care units. They exist in more populated areas and are accepted by workers as the best way to accomplish the diagnosis of occupational diseases. There is important participation by the trade unions in the management of these reference centres. For 30 years now employers organisations have also kept specialised services for safety and occupational health. Although they are better equipped they are less well used by the workers than the CRSTs. At the federal level, activities concerned with occupational health are connected to three ministries: Labour, Health and Social Security. The Ministry of Labour enacts legislation on hygiene, safety and occupational medicine, performs inspections through its regional units and runs a number of research projects. The Ministry of Health provides medical care for workers injured or affected by occupational diseases and also has surveillance programmes for certain occupational diseases. The Ministry of Social Security provides rehabilitation and compensation for registered workers. In spite of a decrease in the number of accidents at work during the past 25 years, working conditions have not improved. Changes in the laws of social security in the 1970s discouraged registration and reporting of occupational injuries and diseases. In consequence death rates due to accidents increased. With the implementation of the CRSTs, the recorded incidence of occupational diseases has risen, not only because of improved diagnosis, but also because of stronger pressure from the unions and better organisation of public services and enterprises. PMID- 9342621 TI - A critical review of epidemiology studies of trichloroethylene and perchloroethylene and risk of renal-cell cancer. AB - The epidemiology studies of trichloroethylene (TCE) and perchloroethylene (PCE) as they relate to risk of renal-cell cancer are critically reviewed. The studies fall into two basic groups: cohort studies of workers who use TCE or PCE and community-based case-control studies. Issues of bias, confounding, and chance are examined in relation to the studies. There is little evidence of an increased risk of renal-cell cancer and exposure to TCE or PCE. The few studies with elevations in risk suffer from important methodologic shortcomings. Although it is virtually impossible using epidemiology data to rule out conclusively a small increase in risk of renal-cell cancer, the totality of epidemiologic evidence clearly does not support a causal association with TCE or PCE. Future studies of these chemicals must include quantitative evidence of exposure and proper methodologic design, be large-scale in nature to detect small increases in risk, and provide a coherent interpretation of all epidemiology data on these solvents and risk of renal-cell cancer. PMID- 9342622 TI - Biological monitoring of workers exposed to low levels of 2-butoxyethanol. AB - OBJECTIVES: (1) To assess the value of urinary butoxyacetic acid (BAA) measurement for the monitoring of workers exposed to low concentration of 2 butoxyethanol (BE); (2) to evaluate the in vivo effect of low occupational BE exposure on the erythrocyte lineage; and (3) to test the possible influence of genetic polymorphism for cytochrome P450 2E1 (CYP 2E1) on urinary BAA excretion rate. METHODS: Thirty-one male workers exposed to BE in a beverage package production plant were examined according to their external (BE) and internal (BAA) solvent exposure. The effect of this exposure on erythrocyte lineage [red blood cell numeration (RBC), hemoglobin (Hb), hematocrit (Htc), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), haptoglobin (Hp), reticulocyte numeration (Ret) and osmotic resistance (OR)], hepatic [aspartate aminotransferase (GOT), alanine aminotransferase (GPT)] and renal [plasmatic creatinine, urinary retinol binding protein (RBP)] parameters was also investigated. DNA purified from whole blood was used for CYP 2E1 genotyping. RESULTS: Average airborne concentration of BE was 2.91 mg/m3 (0.59 ppm) with a standard deviation of 1.30 mg/m3 (0.27 ppm). There was a relatively good correlation between external and internal exposure estimated by measuring BAA in post-shift urine samples (average 10.4 mg/g creatinine; r = 0.55; P = 0.0012). Compared with a matched control group (n = 21) exposed workers had a statistically significant decrease (3.3%; P = 0.03) in Hct while MCHC was increased (2.1%; P = 0.02). No significant difference was observed either in other erythroid parameters or in hepatic and renal biomarkers. One exposed individual exhibited a mutant allele with increased cytochrome P450 oxidative activity which coincided with a very low urinary BAA excretion. CONCLUSIONS: Single determination of BAA in post-shift urine samples can be used to assess exposure to low levels of BE. A slight but significant effect on erythroid parameters suggesting membrane damage was observed in exposed workers. The influence of the genetic polymorphism for CYP 2E1 deserves further investigation for the interpretation of urinary BAA measurements. PMID- 9342623 TI - Assessment of biological chromium among stainless steel and mild steel welders in relation to welding processes. AB - Air and biological monitoring were used for assessing external and internal chromium exposure among 116 stainless steel welders (SS welders) using manual metal arc (MMA), metal inert gas (MIG) and tungsten inert gas (TIG) welding processes (MMA: n = 57; MIG: n = 37; TIG: n = 22) and 30 mild steel welders (MS welders) using MMA and MIG welding processes (MMA: n = 14; MIG: n = 16). The levels of atmospheric total chromium were evaluated after personal air monitoring. The mean values for the different groups of SS welders were 201 micrograms/m3 (MMA) and 185 micrograms/m3 (MIG), 52 micrograms/m3 (TIG) and for MS welders 8.1 micrograms/m3 (MMA) and 7.3 micrograms/m3 (MIG). The curve of cumulative frequency distribution from biological monitoring among SS welders showed chromium geometric mean concentrations in whole blood of 3.6 micrograms/l (95th percentile = 19.9), in plasma of 3.3 micrograms/l (95th percentile = 21.0) and in urine samples of 6.2 micrograms/l (95th percentile = 58.0). Among MS welders, mean values in whole blood and plasma were rather more scattered (1.8 micrograms/l, 95th percentile = 9.3 and 1.3 micrograms/l, 95th percentile = 8.4, respectively) and in urine the value was 2.4 micrograms/l (95th percentile = 13.3). The analysis of variance of chromium concentrations in plasma previously showed a metal effect (F = 29.7, P < 0.001), a process effect (F = 22.2, P < 0.0001) but no metal-process interaction (F = 1.3, P = 0.25). Concerning urinary chromium concentration, the analysis of variance also showed a metal effect (F = 30, P < 0.0001), a process effect (F = 72, P < 0.0001) as well as a metal-process interaction (F = 13.2, P = 0.0004). Throughout the study we noted any significant differences between smokers and non-smokers among welders. Taking in account the relationships between chromium concentrations in whole, plasma or urine and the different welding process. MMA-SS is definitely different from other processes because the biological values are clearly higher. These higher levels are due to the very significant concentrations of total soluble chromium, mainly hexavalent chromium, in welding fumes. PMID- 9342624 TI - Cadmium in the blood and seminal fluid of nonoccupationally exposed adult male subjects with regard to smoking habits. AB - Blood cadmium (B-Cd) and seminal fluid cadmium (Sf-Cd) were measured in 120 adult male subjects not occupationally exposed to cadmium (Cd), comprising 42 nonsmokers (including nine former smokers) and 78 smokers. The respective median and range values were: 0.46 (0.19-1.49) microgram/l of B-Cd and 0.54 (0.17-1.67) microgram/l of Sf-Cd in nonsmokers, and 4.33 (0.49-13.33) micrograms/l of B-Cd and 0.85 (0.29-3.56) microgram/l of Sf-Cd in smokers. Both indicators showed a highly significant difference in Cd exposure between the groups (P < 0.0001), although the increase in B-Cd was considerably more pronounced than that of Sf-Cd in smokers compared with nonsmokers. The results suggest a nonlinear relationship (log Sf-Cd/log B-Cd: r = 0.501, P < 0.0001), rather than linear relationship (Sf Cd/B-Cd: r = 0.430, P < 0.0001), between the indicators. Significant correlations were found between smoking habits, i.e., the number of cigarettes per day, and an increase in B-Cd in smokers (r = 0.296, P < 0.01) and in all 120 subjects (r = 0.685, P < 0.0001), as well as between smoking habits and an increase in Sf-Cd in smokers (r = 0.378, P < 0.001) and in all 120 subjects (r = 0.488, P < 0.0001). Both indicators are necessary for evaluation of individual internal Cd dose, since they appear to differ in reflecting recent and long-term cumulative Cd exposure and/or the amount of Cd at the site(s) of its effect(s) in the body. PMID- 9342625 TI - Electrophysiological investigation of central, peripheral and autonomic nerve function in workers with long-term low-level exposure to carbon disulphide in the viscose industry. AB - OBJECTIVE: Neurotoxicity of carbon disulphide (CS2) is well known. The air concentration at the workplace at which such adverse effects can first be observed is the subject of controversial discussion. METHODS: In a cross sectional study on CS2-exposed workers peripheral motor and sensory nerve conduction studies, somatosensory evoked potentials, thermotesting and investigation of forced respiration sinus arrythmia have been carried out. The data from 222 workers exposed to CS2 in the viscose industry were evaluated and compared with data from 191 employees from the same factory with similar physical and psychological stress factors but without detectable occupational contact to neurotoxic substances. Median exposure to CS2 was below the currently valid occupational-medical threshold limit value (MAK-value) of 10 ppm. Multiple linear or multiple logistic regression analysis was used to check for statistical differences. RESULTS: Binary evaluation (comparison of exposed persons versus controls after multiple linear regression) revealed a slightly lower value in the exposed group for the motor nerve conduction velocity (MNCV, -0.76 m/s, median 48 m/s), but a long way from pathological thresholds. No dose-response relationship could be found within the exposed group for any evaluation criteria of CS2 exposure. Somatosensory evoked potentials, thermotesting and analysis of heart rate variability yielded no indication of a neurotoxic effect of CS2. CONCLUSION: Isolated decrease of MNCV in binary evaluation is, with regard to the known mechanism of CS2-neurotoxicity and the lack of a dose-response relationship, obviously not due to toxic effects. We interpret our results as showing that an adverse effect of carbon disulphide at the exposure ranges found was not detectable in the exposed group. PMID- 9342626 TI - Elimination of 1-hydroxypyrene after human volunteer exposure to polycyclic aromatic hydrocarbons. AB - The aim of this study was to estimate the kinetics of 1-hydroxypyrene (1-HP) elimination after inhalation exposure to polycyclic aromatic hydrocarbons (PAHs). Samples of inhaled and exhaled air were collected on glass fiber filters backed with tubes filled with Amberlit XAD-2 resin. The filters were extracted by cyclohexane and Amberlit--by acetonitrile. Extracts for the determination of pyrene and benzo[a]pyrene (B[a]P) concentrations were analyzed by high performance liquid chromatography (HPLC). 1-Hydroxypyrene in urine was determined after its preconcentration on a C-18 column (solid phase extraction method) using the same analytical technique. Five male volunteers were exposed for 6 h (two times, with a 1-month interval) to a PAH mixture at an aluminium plant. The volunteers were breathing at rest through facial mask equipped with a 1000-ml compensation container which allows collection of the exhaled air. Inhaled air samples were collected in the breathing zone of each volunteer. Urine samples were collected until the 71st hour after the onset of exposure. The average respiratory retention of pyrene was found to be 61%. The 1-HP elimination process could be described by one-compartment model with the half-live of 9.8 hour (95% CI 7.9-11.7 h). The simulation of 1-HP elimination in urine during a working week (4 days) indicates that the balance between absorption and elimination is achieved at the end of the second day. PMID- 9342627 TI - Assessment of exposure to carcinogenic N-nitrosamines in the rubber industry. AB - Exposures to volatile nitrosamines were measured at 24 rubber manufacturing plants from 1992 to 1995. A total of 709 exposure measurements were taken in general areas or personal breathing zones to estimate exposure according to production types (seals, joints, tyres, gloves, etc.) and production steps, from mixing to storage. Five different nitrosamines were identified. N Nitrosodimethylamine is the most frequently encountered nitrosamine and represents the most important fraction of the total nitrosamine concentration measured in a given sample. This fact is consistent with the use of rubber additives containing corresponding amine precursors. One hundred and forty-one of the 709 values exceeded the German target value (TRK) of 2.5 micrograms/m3 for all nitrosamines present from rubber vulcanisation, the only available standard for occupational nitrosamine exposures. The salt bath curing process generates particularly high nitrosamine levels, 90% of the 96 measurements being over the TRK, with many values exceeding 20 micrograms/m3. The reasons why the TRK is exceeded are generally well identified. To reduce nitrosamine emission levels it would be advisable to eliminate nitrogen oxide sources, principally by using a process other than salt bath curing, and to develop different rubber stocks that do not contain secondary aliphatic amine functional groups ("safe amines"). PMID- 9342628 TI - Nasal septum lesions and lung function in workers exposed to chromic acid in electroplating factories. AB - The objective of this study was to compare workers from chromium, nickel chromium, and zinc electroplating factories with regards to nasal septum lesions and lung function. Also investigated was the relationship between chromium levels in air and urine. A total of 189 workers from 11 electroplating factories (three chromium, six nickel-chromium, two zinc) were chosen from central Taiwan. All subjects were interviewed by constructed-questionnaire, given a nasal examination by a certified otolaryngologist and a lung function test. In the chromium factories 30.8% of the workers showed evidence of nasal septum perforations and 38.5% showed evidence of nasal septum ulcers. A Mantel extension test for trend showed that workers in the chromium factories were 31.7 times more likely to experience nasal ulcers than nickel-chromium and zinc factory workers. Those who worked in the electroplating tank area were 4.2 times more likely to experience ulcers and those with over 9 years' experience were 30.8 times more likely. A comparison of lung function adjusted for age, gender and smoking habit among workers showed that vital capacity (VC), forced vital capacity (FVC), and forced expiratory volume in 1 s (FEV1) were all significantly decreased among chromium factory workers. Because the results showed that the workers' health is being severely damaged by the harmful environment of chromium electroplating factories, the authors wish to suggest improvements in the workplace are vitally needed to ensure the safety of the workers. PMID- 9342629 TI - Differences in hand and foot psychomotor speed among 18 pairs of monozygotic twins discordant for lifelong vehicular driving. AB - The purpose of this study was to examine driving as a determinant of hand and foot psychomotor reaction times. Visual simple and choice hand and foot psychomotor reaction times were measured. The occupational driving contrast was determined by an interview reviewing every job held during each subject's lifetime. Comparison was made of psychomotor speed among 18 pairs of 39- to 62 year-old monozygotic male twins discordant for lifelong occupational driving. The mean discordance was the equivalent of 16 years of full-time driving. The twins who drove more tended to have slower hand simple and choice reaction times, although only the difference in hand-choice decision time was statistically significant (32 ms, P < 0.05). The drivers also had slower ipsilateral foot choice decision times (21 ms, P < 0.01), but on average they had faster reaction times in 8 of the 12 ipsilateral and contralateral foot measurements. The slightly longer decision times could be related to some general harmful effects of driving, possibly whole-body vibration. Faster foot movement times of drivers may be affected by practice effects of rapid lower-extremity movements in driving. PMID- 9342630 TI - Extensive lead exposure in children living in an area with production of lead glazed tiles in the Ecuadorian Andes. AB - We have determined the concentrations of lead (Pb), cadmium (Cd), and mercury (Hg) in the blood of children living in two Andean villages in Ecuador with many family-owned cottage-type industries using Pb from discarded car batteries and occasionally, utility batteries containing Cd and Hg for the production of glazed tiles. The battery metals are ground together with water to a suspension, which is applied manually onto the tiles and then fused at about 1,200 degrees C in sawdust-fired kilns. Children aged 4-15 years were recruited from the schools with the assistance of the school-teachers. Children from homes with and without tile-glazing activities were to be included. Blood metal concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS). The children had extremely high blood lead concentrations (B-Pb), which ranged between 100 and 1,100 micrograms/l (median 510 micrograms/l, n = 82). Children from families engaged in tile-glazing production had significantly higher B-Pb (median 600 micrograms/l) than those living in homes with no such activity (median 210 micrograms/l), although the B-Pb of the latter were nonetheless clearly elevated. B-Cd and B-Hg were low (medians 0.25 microgram Cd/l and 1.6 micrograms Hg/l, respectively), indicating that the exposure from utility batteries containing Cd and Hg was low. The blood hemoglobin concentrations decreased significantly with rising B-Pb, indicating an effect on the heme synthesis. This was supported by a marked increase in the blood concentration of protoporphyrins with increasing B Pb. It can be concluded that children from families with cottage industries producing glazed tiles are at risk for severe health effects due to high lead exposure. PMID- 9342631 TI - Cost-effectiveness of tumor marker detection in cancer patients. PMID- 9342632 TI - Detection of the circulating lung cancer marker LCAP with a new monoclonal antibody TRD-L1. AB - Lung Cancer Associated Protein (LCAP) is a high molecular weight glycoprotein defined by the monoclonal antibody (MAb) DF-L1 prepared against a primary adenocarcinoma of the lung. Previous studies have demonstrated that LCAP circulates at elevated levels in patients with lung cancer. However, a suitable assay for monitoring LCAP levels has not been available. The present work describes the development of a double-determinant LCAP assay using MAb TRD-L1 as the capture antibody and MAb DF-L1 as the tracer. In 60 normal subjects, the mean LCAP level was 4.8 U/ml with 2 (3.3%) subjects having values > 12 U/ml (mean + 2SDS). By contrast, 37 of 67 (55.2%) patients with lung cancer had LCAP levels > 12 U/ml. Moreover, only 14 of 203 (6.9%) patients with benign lung disease had elevated levels. LCAP levels were most commonly elevated (62.7%) in patients with adenocarcinoma of the lung and with advanced disease. These results indicate that LCAP as detected by MAb TRD-L1 is a potentially useful marker for the evaluation of patients with lung cancer. PMID- 9342633 TI - Serum gonadotropin levels in patients with germ-cell tumors of the testis: interrelations, possible cross-reactions and interpretation of beta-HCG level. AB - The concentration of FSH, LH, LTH, testosterone and beta-hCG was estimated in 177 serum samples from 86 patients with malignant germ-cell tumors of the testis. The objectives of the investigation were the following: the detection of interrelations of hypophyseal gonadotropins at different beta-hCG levels; the determination of the significance of borderline values of beta-hCG; the analysis of the effect of elevated concentrations of beta-hCG on pituitary gonadotropins: the detection of possible cross-reactions during gonadotropin determinations. The RIA method was used to estimate levels of three gonadotropins. The results revealed that there was no cross-reaction between FSH and beta-hCG at RIA assays. When the serum level of beta-hCG of tumor origin exceeded 100 U/l a subtotal inhibition of FSH secretion was observed. Pathologically increased values of beta hCG were found not only in serum with subnormal-FSH levels, but also when FSH levels were excessively elevated (exceeding 50 U/l). In the latter case the elevated beta-hCG levels could possibly be the consequence of the secretion of beta subunits by the hypophysis or a cross-reaction with LH, and not of a tumor. With values of beta-hCG over 100 U/l cross-reaction with LH occurs, so the true LH levels cannot be assessed. For an adequate interpretation of elevated values of beta-hCG in the serum (i.e. whether they are tumor-derived or not), it is necessary to have values of FSH from the same serum sample. PMID- 9342634 TI - Role of sialic acid and alkaline DNase in breast cancer. AB - Serum levels of sialic acid and alkaline DNase (ADA) were analysed in 495 blood samples collected from 170 breast cancer patients before and during/after anticancer treatment. Fifty-six healthy females were included in the study to define the cutoff values. The markers were analysed by highly sensitive spectrophotometric methods. Statistical evaluation of the data was done using Student's 't' test, paired 't' test and ROC curve analysis. The total sialic acid (TSA), lipid bound sialic acid (LSA) and ADA in sera of untreated breast cancer patients were significantly higher than in controls. ROC curve analysis revealed TSA and LSA to be useful markers for diagnosis of breast cancer. Serum levels of TSA and LSA were significantly decreased in complete responders as compared to their pretreatment values. The pretreatment ADA values showed much individual variation. However, responders showed higher levels of ADA than untreated patients. In nonresponders the values of the biomarkers were comparable with pretreatment levels. The study suggested that TSA and LSA can be helpful in the diagnosis of breast cancer. All three markers can be used for assessment of response to anticancer treatment in breast cancer patients. PMID- 9342635 TI - Histopathological and prognostic evaluation of immunohistochemical findings in colorectal cancer. AB - Many immunohistochemical studies have investigated the relationship between immunohistochemical characteristics and histopathological findings in colorectal tumors. One of the most extensively studied markers has been tissue CEA, although the prognostic significance of this and other antigens is still uncertain. The authors report results relative to three tumoral antigens (carcinoembryonic antigen, CEA; tissue polypeptide antigen. TPA, and carbohydrate antigen 19-9, CA 19-9) determined by immunohistochemical methods in tissue samples of 52 colorectal carcinomas. The relationship between the immunohistochemical characteristics of the neoplasms and the clinicopathologic parameters, as well as their influence on the prognosis of the patients, were examined. Positive CEA reaction has a significant relationship with grade of differentiation of the tumor while diffuse cellular expression of this antigen often indicates neoplasms extending beyond the intestinal wall and invading the lymph vessels. The number of tissue antigens expressed is significantly related to the extent of tumor spread through the intestinal wall. A greater incidence of recurrence and shorter disease-free interval and survival were observed in neoplasms that expressed tissue TPA antigen or more than one tissue antigens. In the present study the latter parameter has demonstrated to have independent prognostic significance for the disease-free interval. Immunohistochemical evaluation of antigens in colorectal carcinoma tissue shows a possible independent prognostic value of the antigenic heterogeneity of tumors, which could be related to their different biological behavior. PMID- 9342636 TI - Interleukin-4 blood concentrations in early and metastatic human solid neoplasms. AB - Blood levels of the immunosuppressive cytokines IL-6 and IL-10 are often abnormally high in patients with advanced cancer. However, since IL-6 and IL-10 may be produced by macrophages and TH2 cells, the evidence of abnormally high values of IL-6 and/or IL-10 may reflect hyperactivation either of the macrophage system or of TH2 cell functions. In contrast, IL-4 is almost completely produced by the TH2 lymphocytes. Therefore, evaluation of IL-4 levels could help to differentiate macrophage from TH2 cell hyperactivation. This study was performed to investigate IL-4 serum levels in a group of cancer patients in relation to the stage of disease and to the secretion of other cytokines. The study included 50 patients, 28 of whom showed distant organ metastases. Lung cancer and gastrointestinal cancers were the most frequent neoplasms in our patients. The control group consisted of 60 healthy subjects. IL-4 was measured by the Elisa method. No patient showed high levels of IL-4. No significant differences were seen between controls and cancer patients, nor between metastatic and non metastatic patients. In addition, no significant differences in IL-4 mean values were found between patients with normal or high levels of IL-6 and IL-10, or between patients with normal or low IL-2 concentrations. This preliminary study seems to exclude cancer-related abnormally high secretion of IL-4. Therefore, the high levels of IL-6 and/or IL-10 often occurring in advanced neoplastic disease would mainly depend on macrophage production. PMID- 9342637 TI - Correlation analysis of alveolar macrophage cytochemical parameters in smoking and pulmonary oncology. AB - Cytochemical examination of alveolar macrophages (AM) obtained by bronchoalveolar lavage (BAL) was performed in healthy volunteers (11 non-smokers and 11 smokers) and in 9 patients with squamous lung carcinoma (all of them smokers or ex smokers) in order to analyze its peculiarities related to the smoking habit and to lung malignancy. Assessment of non-specific esterases: alpha-naphthyl acetate esterase (ANAE) and butyrate esterase (BUT), chloroacetate esterase (CHL), acid phosphatase (AcP), intracellular glycogen (PAS reaction), lipids (Sudan black B reaction-SBB) and iron (Perl's reaction) was performed by a semiquantitative cytochemical method (1). A significant correlation was obtained between BUT and stage of squamous lung carcinoma (varying between I and IV) (r = 0.52, p < 0.05). There was a correlation between BUT and Perl's in healthy controls (r = 0.76, p < 0.05). The same type of correlation was observed in control smokers (r = 0.64, p < 0.05), in addition to a correlation between CHL and AcP (r = 0.69, p < 0.05). There was no significant BUT/Perl's correlation in patients with squamous cell lung carcinoma (r = 0.23, p > 0.05), but significant AcP/CHL correlation as was observed in control smokers (r = 0.73, p < 0.05), and a "new" type of correlation was shown to exist between ANAE and SBB (r = 0.77, p < 0.05). In spite of the unresolved nature of lung cancer, correlation analysis of cytochemical parameters in AM might have an important part in the analysis of their relative contribution to the development of smoking-related disorders and lung malignancies. PMID- 9342638 TI - Serum free beta-hCG, a biomarker for bladder carcinoma patients? PMID- 9342639 TI - The effects of blood pressure resting level and lability on cardiovascular reactions to laboratory stress. AB - Evidence that individuals with elevated resting blood pressures display excessive cardiovascular reactions to laboratory stress is regarded as implicating excessive reactivity in the pathogenesis of hypertension. However, it remains possible that this relationship is artifactual, in that resting blood pressure levels and cardiovascular reactions might both reflect intrinsic cardiovascular lability. To examine this possibility, blood pressure and heart rate were measured at rest and in response to an active laboratory stressor in 1259 men. Systolic blood pressure and heart rate reactions to the stressor were predicted by resting blood pressure. Although cardiovascular lability showed some association with both reactivity and resting blood pressure level, the resting blood pressure level-reactivity relationship survived statistical adjustment for such associations. Accordingly, the excessive cardiovascular reactions characteristic of individuals with elevated resting blood pressure would not appear to be explainable, to any substantial extent, by lability effects. PMID- 9342640 TI - The stress of Stroop performance: physiological and emotional responses to color word interference, task pacing, and pacing speed. AB - Heart rate, frequency of skin conductance responses, and self-reported anxiety were measured during performance of a computer version of the Stroop Color-Word Interference Test, and during a non-conflicting control task involving the color naming of color patches. Stroop and control stimuli were presented individually in order to vary task pacing. Subjects (N = 48) were divided into three groups assigned to self-paced, externally-paced, and fast externally-paced conditions. Performance data revealed that the relative proportion of speed and accuracy reductions which resulted from the Stroop interference varied according to task pacing and pacing speed. Stroop performance was accompanied by heightened HR levels which were sustained throughout the series. State-Anxiety scores increased after both tasks, but only among subjects who completed a large number of trials, i.e. subjects in the self-paced and fast externally-paced groups. Skin conductance responses only varied according to task order and time within series, irrespective of Stroop interference or task pacing. Overall, the results remained in accordance with an effort account of the relationship between attention and cardiac activity. They also provided indications on how the Stroop test may act as an efficient laboratory stressor. PMID- 9342641 TI - Shifts in blood volume alter the perception of posture. AB - Recent experiments have shown that somatic graviceptors exist in humans. Traditionally, extravestibular gravity information has been thought to originate from mechanoreceptors in the joints, muscles and skin. Experiments with normal, paraplegic and nephrectomized subjects revealed that the kidneys and the cardiovascular system are involved in providing truncal gravity information. The present study intends to determine the influence of shifts in body fluid, especially of the distribution of blood along the subjects' spinal (Z-) axis, on the perception of posture. To this end, the distribution of body fluids was altered by means of the technique of lower body negative and positive pressure (LBNP and LBPP). LBNP leads to venous pooling of blood in the legs, whereas LBPP prevents venous blood from pooling, increasing central volume. Changes in blood distribution were measured by segmental impedance cardiography for four body segments: the upper torso (thoracic cavity), lower torso (abdominal and pelvic region), thigh and calf. Seventeen healthy subjects (mean age: 27.3 years) participated in the experiment. They were positioned on the side (right-ear-down head position) on a tilt table which the subjects and the experimenter could tilt via remote control around an axis parallel to the subjects' visual (X-) axis. The experimenter set the initial tilt in total darkness to arbitrary angles while strictly alternating between head-up and head-down tilts. Subjects were then asked to rotate the board until they felt they were in a horizontal posture. Means and variances of eight pairs of settings were taken as a measure of the subjective horizontal posture (SHP). During LBNP (-30 mmHg), subjects perceived being tilted head-up, whereas LBPP (+30 mmHg) led them to feel tilted head-down. The results corroborate the hypothesis of an effect of the blood's mass on graviception and also indicate supplementary contributions of other visceral afferences. PMID- 9342642 TI - Optimal foetal growth in the reduction of learning and behaviour disorder and prevention of sudden infant death (SIDS) after the first month. AB - A theory is presented that a diet low in polyunsaturated fatty acids (PUFA) in the third trimester of pregnancy may delay myelination and brain maturation. This may underpin learning and behaviour disorders and sudden infant death (SIDS) after the first month, conditions that are associated with lower than average birthweight. Epidemiological evidence is reviewed showing an inverse relation between the proportion of heavy newborns (> 3500 g) and infant mortality rate. Some countries with a lower proportion of heavy newborns despite equally high standards of living and medical care have higher post-neonatal death rates. The higher rates are solely due to SIDS which has a peak mortality within 80-100 days. It is hypothesised that as this is a time when myelination peaks, SIDS may be due to maturational delay. Evidence of subtle CNS changes in brainstem structures and in the neuromuscular system supports an instability in brainstem control systems. Moderate iatrogenic dietary restriction predominates today, but a rising number of women favour a low-caloric low-fat diet especially in the third trimester when the foetus is most susceptible. This may lead to a depressed birthweight, delayed somatic growth and neuronal maturation, such as is observed in SIDS victims. The majority exposed to suboptimal conditions survive, but a few suffer SIDS; confirming post-neonatal susceptibility. Many, especially males, present minor CNS signs and learning/behaviour disorders that could be the sequelae of repeated hypoxic episodes, such as recorded in more than 80% of SIDS victims. To reduce learning/behaviour disorders and prevent death from SIDS after the first month, it is necessary to ensure optimal development by promoting foetal growth. It is advised to avoid unnecessary dieting and to favour a diet high in PUFAs, thus prolonging pregnancy and so increasing birthweight. PMID- 9342643 TI - Individual differences in auditory middle latency responses in elderly adults and patients with Alzheimer's disease. AB - Previous research has suggested that the Pb component of the middle-latency auditory evoked response (MLAER) is differentially abnormal in patients with Alzheimer's disease relative to control subjects. In the present study, this putative abnormality was examined in vertex-recorded MLAERs elicited by monaural stimulation in 14 patients with Alzheimer's disease (six females) and 22 age matched control subjects (10 females). A sex difference in Pb elicitation was revealed in control subjects; Pb area was twice as large in females than males (P < 0.05). Pb and Pa amplitudes and latencies did not differ between male and female control subjects. Comparisons of Pb between patients and controls were conducted within each sex. There was no main effect of group on Pb area, amplitude, or latency, Pa amplitude was significantly larger in patients than control subjects; there was no group difference in Pa latency. This study did not replicate previous reports of differences in Pb between patients with Alzheimer's disease and elderly control subjects. We demonstrated that Pb elicitation may be unreliable in elderly control subjects and found evidence of a possible sex difference. The effects of inter-subject variables (e.g. age, sex) must be understood more fully before MLAERs can be exploited as meaningful markers of brain dysfunction. PMID- 9342644 TI - Electrodermal relationships with personality measures of psychosis-proneness in psychotic and normal subjects. AB - The relationships between several indices of psychosis proneness and bilateral electrodermal orienting were examined in a large group of normal subjects and a heterogeneous group of schizophrenic patients. In the normal group, the Cognitive Disorganisation scale was characterised by shorter latencies and more irregular patterns of responding that may indicate greater disinhibitory processing in the regulation of attentional mechanisms. The Unusual Experiences scale and paranoia (both measures of positive or 'active' schizotypy) were associated with a predominance of left hemisphere influences in normal female controls. High Introvertive Anhedonia scores predicted non-responding status in schizophrenic patients and hypo-responsivity in normal controls in support of some previous studies. Overall the results support the specificity and validity of different measures of psychosis proneness and the utility of psychophysiological investigation beyond the psychotic disorders themselves. PMID- 9342645 TI - Association of an oculomotor delayed response task and the Wisconsin Card Sort Test in schizophrenic patients. AB - The relationship between a widely-used neuropsychological test of frontal lobe function, the Wisconsin Card Sort Test, and spatial working memory, as assessed by the oculomotor delayed response task, was examined in schizophrenic patients. Schizophrenic patients were impaired compared with bipolar and normal control subjects on both tasks and their working memory performance was significantly correlated with their WCST measures. The spatial working memory and the WCST deficits in schizophrenia may reflect disrupted circuitry mediated by the dorsolateral prefrontal system. PMID- 9342646 TI - Anxiety and respiratory patterns: their relationship during mental stress and physical load. AB - In the present study we investigated the effect of mental stress on respiration using unpleasant sounds. To compare the center output of each stimuli, subjects took part in one session divided into two phases: a mental stress test and a physical loading test. The purpose of this study was not only to investigate ventilatory response in emotions caused by mental stress and physical load, but also to determine the relationship between respiratory pattern and personality. Ten normal subjects were measured for VE (minute ventilation), VT (tidal volume), RR (respiratory rate), Vo2 (O2 consumption), Vco2 (CO2 production) and FETco2 (end-tidal CO2 concentration) on a breath-by-breath basis; the subjects were given Spielberger's State Trait Anxiety Inventory (STAI) before beginning this experiment. Unpleasant emotions caused by mental stress altered the breathing pattern. VE increase was achieved by the combination of VT and RR disregarding the subjects' personality. However, subjects with high anxiety RR increased more than VT resulting in a positive correlation between the trait anxiety score and RR. We found that a dominant RR increase was observed not only in the mental stress test but also in the physical loading test. In the physical load, there was a positive correlation between the state anxiety score and RR. These results indicate that respiratory patterns are related to personality anxiety. These findings may provide important evidence relating respiratory function to psychological aspects. PMID- 9342647 TI - Menstrual cycle, blood pressure and ischemic pain sensitivity in women: a preliminary investigation. AB - Eleven women were tested twice for ischemic pain sensitivity; once during their follicular phase (Days 4-9) and once during their mid-late luteal phase (5-10 days after ovulation) of a confirmed ovulatory cycle. Additionally, in order to examine blood pressure-related hypoalgesic effects, each had 3-4 clinic blood pressures determined during an initial screening interview and each also completed a daily symptom calendar for one complete menstrual cycle prior to testing in order to investigate relationships between 'real life' symptomatology and laboratory-induced pain sensitivity. Results revealed significantly shorter pain tolerance times and marginally shorter pain threshold times in the luteal vs. follicular phase, while verbal descriptors of pain intensity (sensory) and pain unpleasantness (affective) did not vary with cycle phase. Clinic blood pressures were positively correlated with pain threshold and tolerance times assessed during both cycle phases. Real-life physical symptom ratings were predictive of laboratory pain intensity ratings during the follicular phase and tended to predict unpleasantness ratings during both phases. These results not only confirm recent reports of greater sensitivity to ischemic pain in women during the luteal phase of their cycle, but extend the literature by demonstrating pressure-related hypoalgesic effects in women during both cycle phases. PMID- 9342648 TI - Phase transition parameters of potential thermosensitive drug release systems based on polymers of N-alkylmethacrylamides. AB - The phase separation and its thermohysteresis in dilute aqueous solutions of polymeric components of potential drug release systems (homopolymers and copolymers of N-isopropylacrylamide, N-isopropylmethacrylamide, N propylmethacrylamide, N-sec-butylmethacrylamide, and N-(2 hydroxypropyl)methacrylamide) was studied, both on heating and cooling. Plots of light transmittance vs temperature were constructed and the parameters characterizing them were correlated with polymer structures. Qualitative information was obtained on the rate of formation of the concentrated phase on heating and its disappearance on cooling. Attention has been drawn to the improper identification of the cloud-point temperature, measured at an arbitrary concentration, with the lower critical solution temperature (LCST) as is frequently found in papers dealing with biomedical applications of thermosensitive polymers. PMID- 9342649 TI - Influence of heparin coating on in vitro bacterial adherence to poly(vinyl chloride) segments. AB - End-point attached, covalently bound heparin has been shown to be effective in preventing activation of the coagulation cascade by biomaterials. Data concerning its possible influence on bacterial attachment and resistance to biomaterial associated infection are, so far, lacking. In the present work, the in vitro adherence of Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli, one isolate of each species, to plain poly(vinyl chloride) (plain PVC) and heparin coated poly(vinyl chloride) (EPA-PVC) segments was compared. Also, the influence of precoating the segments with human normal plasma for 2 h was studied. 35S-Methionine was used to radiolabel bacteria. The segments were exposed to bacterial suspensions of approximately 10(7) colony forming units (CFU) per milliliter at 37 degrees C for 0.5-6 h. Following repeated washing in phosphate buffered saline (PBS), radioactivity associated with the segments was measured. Plain PVC as compared to EPA-PVC bound significantly more cells of all three tested species. Plasma precoating significantly decreased adherence of the tested species to plain PVC but did not affect the binding to EPA-PVC. However, after precoating with human plasma, EPA-PVC compared to plain PVC showed a higher binding of S. aureus which might possibly be due to bridging effects of fibronectin or other plasma proteins, interacting with S. aureus. PMID- 9342650 TI - Fabrication of microspheres using blends of poly(ethylene adipate) and poly(ethylene adipate)/poly(hydroxybutyrate-hydroxyvalerate) with poly(caprolactone): incorporation and release of bovine serum albumin. AB - Spherical microspheres composed of polymer blends 80:20 PEAD/PCL II and 40:40:20 PEAD/P(HB-HV)/PCL II containing a range of BSA loadings have been fabricated using a single emulsion technique with solvent evaporation. 80:20 PEAD/PCL II microspheres had smooth surfaces while 40:40:20 PEAD/P(HB-HV)/PCL II microspheres consisted of a mixture of smooth surfaced, microporous and macroporous microsphere fractions. Irrespective of fabrication polymer, microspheres were produced in high yield (> 75%) and BSA incorporation had no significant effect on microsphere size distribution which ranged from 0.6 to 5 microns and from 2.1 to 50 microns for 80:20 PEAD/PCL II and 40:40:20 PEAD/P(HB-HV)/PCL II microspheres, respectively. The loss of BSA by partitioning into the aqueous phase resulted in low encapsulation efficiencies (< 14.5%). BSA release increased significantly with the-oretical percentage loading but the relationship could not be confirmed when the total cumulative release of BSA was expressed as a percentage of the actual total BSA incorporated. Significant BSA release could be detected for up to 26 days. PMID- 9342652 TI - Blood compatibility of polypropylene surfaces in relation to the crystalline amorphous microstructure. AB - Blood-contacting properties of polypropylene surfaces with different crystalline states at the surface layer were examined in terms of plasma protein adsorption and changes in cytoplasmic free Ca2+ levels in platelets. Though the wettability of polypropylene surfaces was almost constantly independent from the surface layer crystallinity and interlamellar spacing, an increase in adhesiveness was observed with decreasing surface layer crystallinity and interlamellar spacing. It is suggested that the surface properties of the sheets varied in relation to the crystalline-amorphous microstructure. Minimum magnitudes in albumin and fibrinogen adsorption were observed on the polypropylene surface with a particular surface layer crystallinity (c. 55 wt%). A decrease in interlamellar spacing resulted in enhancing albumin adsorption and diminishing fibrinogen adsorption. Transient phenomena in plasma protein adsorption were observed on their surfaces with a plasma concentration. It is considered that the polypropylene surface with a particular crystalline-amorphous microstructure reduces the denaturation of adsorbed proteins. An increase in cytoplasmic free Ca2+ levels in platelets was prevented at the polypropylene surface with a surface layer crystallinity of 55 wt%: the particular crystalline-amorphous microstructure of such apolar surfaces as polypropylenes acts to reduce platelet activation. Thus, it is concluded that the blood compatibility of polypropylene surfaces is greatly improved by controlling a crystalline-amorphous microstructure at the surface layer. PMID- 9342651 TI - Hemocompatibility studies of surface-treated polyurethane-based chronic indwelling catheters. AB - The objectives of this research were to evaluate and compare the interactions of several polyurethane-based central venous catheter materials with blood. Specifically, measurements of fibrinogen adsorption, platelet adhesion, kallikrein generation, and fibrinopeptide A (FPA) release were performed. The catheter materials examined in this study included: platinum-cured, 50 shore A durometer, barium sulfate-filled, silicone (SI); Tecoflex EG85A-B20 polyurethane (PU); PU catheters whose outer surface had been impregnated with ion beam deposited silver atoms (AgI and AgII); PU catheters coated with a hydrophilic, polyacrylic acid polymer (UC); PU catheters coated with an air-cured PTFE emulsion (CS); and PU catheters coated with an aminofunctional dimethylsiloxane copolymer (JG). The time course of fibrinogen adsorption from plasma to the SI, JG, PU, and CS materials was similar, with CS exhibiting the least amount of adsorbed fibrinogen after 1 h (65 +/- 4.7 ng cm-2) and PU the greatest (144 +/- 16.5 ng cm-2). After 90 min of contact, AgI and AgII exhibited the greatest number of adherent platelets, levels that were approximately two to three times higher than those on the other catheter materials. With the exception of UC and PU, which caused kallikrein generation levels approximately half that of the positive (glass) control, little kallikrein formation was observed for any of the materials relative to the negative control. Finally, FPA generation was greatest using the SI, CS, and PU materials, with the latter causing the production of almost four times the amount of FPA as the negative control. This preliminary assessment of the hemocompatibility of the various catheters suggests that the surface treatments did not adversely affect their interactions with blood components; further investigations of these materials are therefore warranted in order to completely characterize their behavior prior to use in clinical situations. PMID- 9342653 TI - Cell activation by the micropatterned surface with settling particles. AB - Surface topography plays an important role in cell orientation and morphogenesis. In this study, we prepared a micropatterned surface with settling particles to obtain more detailed information about the cell recognition against the microstructured surface. Core-shell type particles having a poly-(N isopropylacrylamide) (polyNIPAM) shell were prepared by seeded polymerization. Particles were settled on a polystyrene (PSt) flat dish by the spinner to prepare a micropatterned surface with settling particles. It could be seen that the polyNIPAM shell shrunk above and swelled below the LCST. For comparison, a thermosensitive flat surface was prepared by the graft polymerization of NIPAM. No morphologic change of cells contacting the both surfaces was observed with either an optical or a scanning electron microscope. Moreover, particles could move or roll on these surfaces when shaking the dishes. The weak interaction between neutrophil-like cells and the micropatterned surface with settling particles or the polyNIPAM-grafted surface was estimated by measurement of active oxygen released by cells. A little release could be observed at both 25 and 35 degrees C. The amount of released active oxygen at 35 degrees C was slightly larger than at 25 degrees C. When the temperature was suddenly changed, the dynamic changes of particle shape and size resulted in the excess release of active oxygen from cells contacting the micropatterned surface with settling particles. Meanwhile no stimulation could be observed in the polyNIPAM-grafted surface even if the temperature is suddenly changed. These results indicate that the micropatterned surface with settling particles can induce the dynamic stimulus at a patterned input mode. PMID- 9342655 TI - Capillary electrophoresis in clinical chemistry. AB - Since its introduction, capillary electrophoresis has diversified, spreading out into different specialized fields covering solutions for almost any analytical questions arising in research laboratories. In the context of clinical chemistry, results must be provided at low costs and in a clinically relevant time frame; however, the attributes which have made capillary electrophoresis such a successful tool in basic research are identical to those attracting clinical laboratories: speed (more efficient, less labor-intensive), low costs (minimal buffer consumption), small sample volume (reduced blood collection volume from patient), increased selectivity (determination of multiple solutes in one run), and versatility (detection of analytes over the wide range of molecular masses and chemical composition). Nevertheless, it should be mentioned that there are still some drawbacks at this stage to be solved in the near future, such as lack of sensitivity for many clinical applications or the constraint to measure in a sequential mode. The aim of this survey is to familiarize clinical chemists, as well as chemists, with a short introduction to capillary electrophoresis, followed by chapters reviewing prominent fields of applications and the latest developments in clinical chemistry. PMID- 9342654 TI - Characterization of partially saturated poly(propylene fumarate) for orthopaedic application. AB - A partially saturated linear polyester based on poly(propylene fumarate) (PPF) was synthesized for potential application in filling skeletal defects. The synthesis was carried out according to a two-step reaction scheme. Propylene glycol and fumaryl chloride were first combined to form an intermediate fumaric diester. The intermediate was then subjected to a transesterification to form the PPF-based polymer. This method allowed for production of a polymer with a number average molecular weight up to 1500 and a polydispersity index of 2.8 and below. The polymeric backbone structure was investigated through the use of FTIR and NMR. Kinetic studies of the transesterification allowed mapping of the molecular weight increase with reaction time. The final product was also characterized by thermal and solubility analysis. PMID- 9342656 TI - New approaches in clinical chemistry: on-line analyte concentration and microreaction capillary electrophoresis for the determination of drugs, metabolic intermediates, and biopolymers in biological fluids. AB - The use of capillary electrophoresis (CE) for clinically relevant assays is attractive since it often presents many advantages over contemporary methods. The small-diameter tubing that holds the separation medium has led to the development of multicapillary instruments, and simultaneous sample analysis. Furthermore, CE is compatible with a wide range of detectors, including UV-Vis, fluorescence, laser-induced fluorescence, electrochemistry, mass spectrometry, radiometric, and more recently nuclear magnetic resonance, and laser-induced circular dichroism systems. Selection of an appropriate detector can yield highly specific analyte detection with good mass sensitivity. Another attractive feature of CE is the low consumption of sample and reagents. However, it is paradoxical that this advantage also leads to severe limitation, namely poor concentration sensitivity. Often high analyte concentrations are required in order to have injection of sufficient material for detection. In this regard, a series of devices that are broadly termed 'analyte concentrators' have been developed for analyte preconcentration on-line with the CE capillary. These devices have been used primarily for non-specific analyte preconcentration using packing material of the C18 type. Alternatively, the use of very specific antibody-containing cartridges and enzyme-immobilized microreactors have been demonstrated. In the current report, we review the likely impact of the technology of capillary electrophoresis and the role of the CE analyte concentrator-microreactor on the analysis of biomolecules, present on complex matrices, in a clinical laboratory. Specific examples of the direct analysis of physiologically-derived fluids and microdialysates are presented, and a personal view of the future of CE in the clinical environment is given. PMID- 9342657 TI - Capillary electrophoresis for pharmacokinetic studies. AB - Different analytical techniques involving capillary electrophoresis for the determination of drugs and metabolites in biological fluids are described. Pharmacokinetic studies carried out using capillary electrophoresis are presented, as well as the in vitro metabolism investigations. The advantages and the limitations of capillary electrophoresis for pharmacokinetic studies are discussed. PMID- 9342658 TI - Guidelines in selecting ligand concentrations for the determination of binding constants by affinity capillary electrophoresis. AB - This study examined various factors that affect the selection of ligand concentrations when using affinity capillary electrophoresis (ACE) for the determination of equilibrium constants in free solution. Two groups of model systems were used in this work: the binding of nitrophenols to alpha- or beta cyclodextrin and the binding of D- or L-tryptophan to human serum albumin (HSA). Both systems gave 1:1 binding behavior in the ACE studies and good fits to previous equations derived to describe the shift in analyte mobility that occurs as the ligand concentration of the running buffer is varied. Some practical factors limiting the range of ligand levels that could be used in such studies included the relative amount of injected analyte, ligand solubility and the ligand's background signal. More fundamental factors included the size of the equilibrium constant for the system being investigated, the relative range of mobilities over which the analyte peak might be observed, the precision of the mobility measurements and the number of analytes present in the sample. Equations and graphs were developed for illustrating each of these latter items and their role in determining the range of ligand concentrations that could be used in ACE binding constant measurements. The results predicted by these equations and graphs showed good agreement with those observed experimentally, and should prove useful in optimizing ACE conditions for other solutes and ligands. PMID- 9342659 TI - Capillary electrophoresis and microdialysis: current technology and applications. AB - Microdialysis sampling has become an important method for the continuous monitoring from an in vivo environment. This technique has been used to monitor many endogenous molecules, such as neurotransmitters, as well as exogenous species such as drug substances. Microdialysis samples have traditionally been analyzed by liquid chromatographic (LC) methods to gain resolution and quantification of the molecules of interest. However, LC separations have a relatively large injection volume requirement which, as a consequence, increases microdialysis sampling times. Capillary electrophoresis (CE), with its very small sample volume requirements and high resolving power, has therefore gained popularity as an alternative to LC. Reviewed here are many of the technologies currently available for CE and examples of how this technique has been effectively applied to the analysis of microdialysis samples. PMID- 9342660 TI - Microdialysis-immunoaffinity capillary electrophoresis studies on neuropeptide induced lymphocyte secretion. AB - A micro-sampling procedure has been developed for studying lymphocyte secretion of biologically important peptides in low cell density cultures. The technique is based on microdialysis recovery of the analytes of interest coupled with immunoaffinity capillary electrophoresis separation of the microdialysis samples and laser-induced fluorescence detection. Although the technique is able to recover secreted materials only at the 5-10 cell level, the detection system has a limit of detection (LOD) in the attomole (10(-18) M) range. This degree of sensitivity indicates that the system has the potential to measure secreted products at the single cell level. An added advantage of this system over other sampling techniques is that the microdialysis probe allows continuous sampling over time. PMID- 9342661 TI - Capillary electrophoresis of recombinant proteins. AB - Many naturally occurring proteins which are used therapeutically have been cloned and expressed in large quantities in bacterial, yeast or mammalian systems. Purification of these proteins by column chromatography generates high purity products with low levels of host protein contaminants. However, isoforms of the desired protein may be present at variable concentrations. Analysis of these variant forms has been enhanced by the utilisation of capillary electrophoresis (CE), a highly efficient, widely applicable technique which is increasingly used in the field of biotechnology. The role of CE in the analysis of recombinant proteins is reviewed with respect to microcharacterisation, comparison of natural and recombinant proteins, separation of mutant or variant forms and analysis of glycoforms. Examples of these applications are described and illustrated with analysis of recombinant human albumin. The rapid development of CE, further enhancing its versatility, and its use with complementary analytical techniques is also discussed. PMID- 9342662 TI - Combination of capillary electrophoresis and matrix-assisted laser desorption ionization mass spectrometry for glycosylation analysis of a human monoclonal anti-Rhesus(D) antibody. AB - Characterization of a human anti-Rhesus(D) monoclonal antibody, developed for the treatment of Rh(D) haemolytic disease of the newborn, was performed. Capillary electrophoresis (CE) has been employed for peptide mapping of the IgG heavy chain and glycopeptide identification. The combination of the high resolution and low solvent consumption of CE and the ultrasensitive detection and precise identification properties of mass spectrometry led to a complete glycosylation analysis of the protein. Glycopeptides were easily isolated from a single injection in a 100 microns i.d. capillary of the preparative CE system and collected for molecular mass determination using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The off-line CE-MS characterization revealed the presence of different oligosaccharides linked to the unique N297-S-T glycosylation site of the IgG heavy chain. The differences between calculated and experimental masses of the glycopeptides suggested the presence of a fucosylated biantennary structure containing one or two galactose units as major oligosaccharide, together with similar species bearing a bisecting N acetylglucosamine. CE conditions were optimized to allow the MS identification of sialylated forms. PMID- 9342663 TI - Detection of a genetic variant, lysine-->glutamic acid at position 372 of human serum albumin, by capillary electrophoresis and structural identification. AB - A genetic variant of human serum albumin (alloalbumin) is detected by capillary electrophoresis (CE). Two albumin peaks, which were in the ratio of approximately one, were clearly separated. One of the peaks had the same migration time as normal albumin (Alb A) and the other (Alb X) had a longer migration time. SDS polyacrylamide gel electrophoresis of CNBr fragments (CB) of Alb X indicated that the amino acid substitution was localized in the CB5 fragment (residue 330-446) of the molecule, because of anomalous migration of CB5 in the gel. The CE mapping of the tryptic peptides from the variant CB5 revealed clearly the existence of a new peptide, and the lack of two normal peptides. The sequence analysis of the variant peptide collected by CE micropreparation showed that the N-terminus of the variant peptide corresponded to that of T49 in Alb A. The substitution site, lysine-->glutamic acid at the position 372, was revealed by sequence determination of the variant peptide purified by reversed-phase HPLC. PMID- 9342664 TI - Analysis of low-molecular-mass proteins in cerebrospinal fluid by sodium dodecyl sulfate capillary gel electrophoresis. AB - Proteins of low molecular (M(r)) in human cerebrospinal fluid (CSF) were analyzed by capillary electrophoresis in a sodium dodecyl sulfate-containing polymer solution. Under the conditions employed, peaks of beta 2-microglobulin (beta MG) (M(r): 11,700), gamma-trace protein (12,300), myelin basic protein (18,000), beta trace protein (beta TP) (23,000 to 30,000) and alpha 1-acid glycoprotein (42,000) were detected on the electropherograms. The concentrations of beta MG and beta TP were determined based on the peak area relative to that of an internal standard, Orange G, which was added at a constant amount as the front marker. It was demonstrated that their levels in CSF change under various pathological conditions in the central nervous system. PMID- 9342665 TI - Sensitive analysis of [D-Pen2,5]enkephalin in rat serum by capillary electrophoresis and laser-induced fluorescence detection. AB - A highly sensitive analytical method based on capillary zone electrophoresis (CZE) coupled with a laser-induced fluorescence (LIF) detector was explored for the analysis of [D-Pen2,5]enkephalin (DPDPE) in rat serum. DPDPE and the internal standard Phe-Leu-Glu-Glu-Ile (P9396) were extracted from serum samples with C18 solid-phase extraction disk cartridges, followed by derivatization with tetramethylrhodamine-5-isothiocyanate (TRITC) isomer G before introduction onto the capillary column. Complete resolution of DPDPE and the internal standard from other serum components was achieved within 20 min on a 140 cm x 50 microns I.D. capillary column with borate buffer (25 mM. pH 8.3). With the current method, it is possible to detect 1.3E-18 mol of DPDPE on column. The results suggest that CZE-LIF is a promising method for the sensitive and specific quantitation of therapeutic peptides in biological matrices. PMID- 9342666 TI - High-performance capillary electrophoresis of a fermentation-derived cyclic peptide analog, animal growth promoter. AB - We have developed HPCE methods for the analysis of sulfomycin (trivial name) and related compounds (code name, crude material = U82127 = I), which is an animal growth promoter derived from a fermentation beer. The crude material, I, isolated from the fermentation consisted of more than 40 components which were not completely separated by conventional HPLC. Thus, as a complementary analysis method, we have optimized HPCE conditions for I using various capillaries including uncoated, coated, and packed using various buffers. The optimized HPCE conditions were obtained with an uncoated fused-silica capillary and a buffer that consisted of 30 mM Tris-tricine, 10 mM SDS, 10 mM NaCl and 20% MeOH, pH 8.0. Using these HPCE conditions, we were able to separate the one main component collected from the HPLC effluent into two or three components. In order to identify the main components of the fermentation product, an off-line HPLC-HPCE MS analysis for I was performed. From the MALDI-TOF-MS results, the separated components collected from HPCE had different molecular masses. Four lots of I samples having different characteristics were also analyzed by HPCE to investigate lot-to-lot differences in peak profiles. The four lots of I were found to have very similar peak profiles. In this paper, I refers to the crude fermentation product and sulfomycin to the purified, major HPLC component of I. PMID- 9342667 TI - Immunoassay for thyroxine (T4) in serum using capillary electrophoresis and micromachined devices. AB - As part of our ongoing effort to develop electrophoretic assay technology for clinical diagnostics, we describe a competitive immunoassay for the determination of serum thyroxine (T4) based on electrophoresis and laser induced fluorescence (LIF). Measurements of total T4 are useful for the clinical evaluation of thyroid function. A fluorescein thyroxine conjugate was utilized in conjunction with a polyclonal antibody preparation as assay reagents. Capillary electrophoresis (CE) conditions tolerant of the direct injection of serum without extraction or other sample preparation steps were developed and used for quantitation of total T4 in serum. We have been exploring the use of micromachined devices with arrays of channels for high assay throughput. Our assay protocol was carried in a microchip format. The results illustrate that gains in speed can be additionally achieved, with the electrophoretic separation of free from bound labelled T4 being performed in about 15 s for serum samples. PMID- 9342668 TI - Temporal analysis of DNA restriction digests by capillary electrophoresis. AB - We demonstrate a facile means for temporal analysis of DNA restriction enzyme digests by capillary electrophoresis-laser-induced fluorescence (CE-LIF) detection. phi X-174 DNA was digested with HaeIII restriction enzyme under conditions that allowed the monitoring of digestion as it proceeded toward completion. Separation by a polymer solution of methylcellulose in a polyacrylamide coated capillary allowed high resolution and a high degree of reproducibility between sequential runs. At pre-selected time intervals an injection of the digest, directly from the reaction mixture, was made. Sensitive detection was achieved by using ethidium bromide as an intercalation dye and allowing intercalation to occur on-column. It is demonstrated that the course of the digestion (i.e., the creation and diminishing of fragment peaks) can be followed using this methodology. Also demonstrated is the ability to use temporal analysis to determine ideal conditions for producing a single cut within a cloning and expression vector (pET3a-PAI-1) which contains 11 potential restriction endonuclease cleavage sites. This initial attempt to follow a restriction digest on-column not only provides meaningful information for the biochemical researcher, but also furthers the use of CE a diagnostic tool for the biochemical laboratory. PMID- 9342669 TI - Direct quantification of HIV-1 RNA by capillary electrophoresis with laser induced fluorescence. AB - HIV-1 RNA was quantitated directly by capillary electrophoresis with laser induced fluorescence (CE-LIF). CE-LIF was used to analyze cellular RNA and various nucleotide complexes. A fluorescently labeled DNA probe (DNA/RNA complex) in conjunction with thiazole orange intercalator was determined to have optimal stability and sensitivity for RNA analysis. Based on this observation, a hybridization method using a HIV-specific fluorescently labeled probe with analysis by CE-LIF was developed. Plasma samples from a HIV-seropositive patient were lysed to obtain RNA, hybridized with the HIV-specific probe and analyzed by CE-LIF. As little as 19 fg (1710 copies per 1 ml of starting plasma) of HIV RNA can be reliably and quantitatively detected. CE-LIF appears to be an efficient and sensitive method to quantitatively analyze RNA from a variety of sources. PMID- 9342670 TI - Non-isocratic capillary electrophoresis for detection of DNA point mutations. AB - Capillary electrophoresis under non-isocratic conditions is reviewed here. In particular, these conditions are elicited by programming the temperature over the run, just as in temperature-programmed gas chromatography. There is at least one major instance in which this technique is particularly useful: detection of DNA point mutations in PCR-amplified DNA fragments. In this case, migration of DNA homo- and hetero-duplexes against a denaturing gradient (either thermal or chemical) is the only technique capable of developing a pattern of four zones, indicative of the presence of a point mutation in genomic DNA, and due to the formation of two homo- (Wt/Wt, M/M, where Wt = wild type and M = mutant) and two hetero- (Wt/M and Wt/M) duplexes, formed by fully denaturing and then re annealing the Wt and M filaments in solution. It is demonstrated that it is possible to obtain such gradients (which do not exist in space, i.e., along the length of the capillary, but in time), simply by producing voltage ramps, which in turn generate temperature ramps from within the capillary lumen, rather than from without, by circulating liquids and thermostats. This technique can be applied to any possible type of mutation, from low- to intermediate to high melters, i.e., from 40 degrees C up to 60-65 degrees C. Examples of separations of point mutations in the cystic fibrosis transmembrane regulator gene and in the beta-globin chain gene are shown. PMID- 9342671 TI - Fractional factorial design optimization of the separation of pilocarpine and its degradation products by capillary electrophoresis. AB - The separation of pilocarpine and its degradation products by micellar electrokinetic capillary chromatography (MECC) has been optimized by using fractional factorial design of the experiments. Critical parameters were identified in a screening design, and an optimization design was used to optimize the separation. The optimal separation method was based on a borate buffer with sodium dodecyl sulfate (SDS). It is concluded that by using fractional factorial design it is possible to improve the separation of pilocarpine, its trans epimer, isopilocarpine and their hydrolysis products, pilocarpic acid and isopilocarpic acid. PMID- 9342672 TI - Separation of components of human globulins by capillary zone electrophoresis using a linear polyacrylamide-coated capillary. AB - Eleven commercially available protein preparation from human serum were subjected to capillary zone electrophoresis (CZE) using a linear polyacrylamide-coated capillary at pH 7.4. Transferrin, complement C3 and C-reactive protein were each separated into one major peak and several minor peaks. alpha 1-Antitrypsin was separated into two major peaks and three minor peaks. alpha 2-HS-glycoprotein showed four major peaks with a leading shoulder. Haptoglobin, alpha 2 macroglobulin, alpha 1-acid-glycoprotein and prealbumin were detected as relatively wide peaks. Ceruloplasmin showed one major peak with notches, and two minor and several notched peaks. Only low density lipoprotein showed no peaks. A mixture of five of the protein preparations was separated into individual components, as well as individual isoforms. When the same mixture was analyzed by CZE using an uncoated capillary, a much poorer resolution was obtained. Application of this CZE system to albumin-depleted serum demonstrated that it is very useful for analyzing globulin components in serum. PMID- 9342673 TI - Use of capillary sodium dodecyl sulfate gel electrophoresis to detect the prion protein extracted from scrapie-infected sheep. AB - Scrapie in sheep and in goats is the prototype of a group of transmissible spongiform encephalopathies (TSE). A feature of these diseases is the accumulation in the brain of rod shaped fibrils that form from an aggregated protein that is a protease-resistant form of a modified normal host cell protein. In this study, we compared SDS gel capillary electrophoresis to conventional SDS PAGE and Western blot to detect the monomer of this aggregated protein. This prion protein was extracted from the sheep brain by homogenizing the brain stem (10%, w/v) in 0.32 M sucrose and by using a series of ultracentrifugation steps and treatment with sodium lauroyl sarcosine and proteinase K. After the final centrifugation step, the pellet was resuspended in 0.01 M Tris pH 7.4 in a volume equivalent to 0.1 ml/g of brain used. This resuspended pellet was treated with 1% SDS and 5% 2-mercaptoethanol and boiled for 10 min. The analysis was done in a Beckman P/ACE 5500 using a SDS gel capillary (eCap SDS14-200 Beckman capillary). In infected sheep brain samples, but not normal sheep, a major peak at a molecular mass of 16.1 kDa and a minor peak with a leading shoulder were observed. Since the molecular mass determined for this protein was lower than that estimated on Western blot (22.4 kDa), a Ferguson plot was made to determine if there were abberations in the molecular mass determination. After correction, the major peak was estimated to be 19.2 kDa. This has a better correlation with that determined by SDS-PAGE and Western blot. The equivalent amount of brain sample in the capillary was approximately 50 micrograms. For Western blot, the amount of brain sample was approximately 20 mg. For this assay, this is approximately 100 times less than that needed for Western blot for sheep samples. PMID- 9342674 TI - Assessment of the quality of dairy products by capillary electrophoresis of milk proteins. AB - This paper presents an overview of existing capillary electrophoretic methods for the study of milk proteins. The main methods of analysis of caseins, whey proteins and peptides are examined with particular attention to their application to the evaluation of the quality of dairy products. Aspects such as the study of protein polymorphism, evaluation of heat treatments, detection of adulteration and assessment of proteolysis are considered in detail. PMID- 9342675 TI - Detection of 8-hydroxydeoxyguanosine in K562 human hematopoietic cells by high performance capillary electrophoresis. AB - A method for the detection of 8-hydroxydeoxyguanosine by high-performance capillary electrophoresis (HPCE) was developed. Separations were performed in an uncoated silica capillary (44 cm x 75 microns i.d.) with a P/ACE system with diode-array detector. The separation of purine deoxynucleosides and 8 hydroxydeoxyguanosine was optimized with regard to pH, temperature, applied potential and hydrodynamic injection time. Optimum conditions were 20 mM borate buffer (pH 9.5), 25 degrees C, 25 kV, 20 s load and detection at 254 nm. This method allowed the detection of 8-hydroxydeoxyguanosine in the presence of a 10(5)-fold higher amount of deoxyguanosine. Isolated nuclei from K562 human hematopoietic cells were treated with 15 mM hydrogen peroxide for 2 h. The nuclei were extensively dialyzed and DNA was isolated, enzymatically hydrolyzed to the deoxynucleosides and analyzed by HPCE. DNA from hydrogen peroxide treated nuclei had a 4-fold higher content of 8-hydroxydeoxyguanosine than untreated controls. HPCE analysis of 8-hydroxydeoxyguanosine is fast and simple. Furthermore, it requires a very small sample volume, which makes it useful for biomedical and clinical applications. PMID- 9342676 TI - Capillary electrophoretic analysis of ethylene dicysteine, a precursor of the radiopharmaceutical 99mTc ethylene dicysteine. AB - L,L-Ethylene dicysteine (EC) is a ligand used in nuclear medicine for the preparation of 99mTc-L,L-EC, a tracer agent for renal function studies. A capillary zone electrophoretic method is described which allows us to monitor the purity of ethylene dicysteine, for which no other suitable method has been reported to date. A fused-silica capillary (L = 70 cm, l = 62 cm, 75 microns i.d.) was used with 10 mM sodium tetraborate, pH 10.0, as the background electrolyte. The voltage applied was 20 kV and the temperature was 25 degrees C. Benzyl alcohol served as the internal standard. The limit of detection was 0.07% and the limit of quantitation was 0.15%. PMID- 9342677 TI - Analysis of trifluoroacetic acid in lyophilized natural products by capillary electrophoresis. AB - A rapid and simple method for the capillary electrophoretic determination of residual trifluoroacetic acid in lyophilized natural products is described. The technique utilizes 2,6-naphthalenedicarboxylic acid as a separation buffer additive providing indirect ultraviolet absorption detection. Using this method, acceptable precision, accuracy, selectivity, range and linearity were achieved. PMID- 9342678 TI - Determination of aminopeptidase X activity in tissues of normo- and hypertensive rats by capillary electrophoresis. AB - Aminopeptidase X, an enzyme that degrades angiotensin I to des-asp-angiotensin I, was determined in the lung, liver, kidney, plasma, endothelium and aortic smooth muscle of the spontaneously hypertensive rat (SHR) and its normotensive control, the Wistar Kyoto rat (WKY). The enzyme activity in the lung, kidney, plasma and endothelium of the SHR was elevated and this supports an earlier suggestion that in certain critical tissues of the SHR, the degradation of angiotensin I is shunted in favour of the des-asp-angiotensin I pathway. In these tissues, the formation of pressor angiotensin II would be curtailed and that of des-asp angiotensin I enhanced. As des-asp-angiotensin I lacks direct vasopressor action, its preferential formation over that of angiotensin II could be a physiological response to the prevailing hypertension in the SHR. PMID- 9342679 TI - Capillary zone electrophoresis separation of tryptophan and its metabolites, including quinolinic acid. AB - Capillary zone electrophoresis with UV absorbance detection was used to separate tryptophan and ten of its metabolites. Run buffers of pH 4.0-10.0 were evaluated for their effect on resolution; a pH 9.6 buffer was found to give optimum separation of all components. Ethylenediaminetetraacetic acid (EDTA), which prevents complexation of some analytes with polyvalent cations, was included in the run buffer to insure good peak shape and reproducible mobilities. The resulting method was used to detect the presence of quinolinic acid in a urine sample. PMID- 9342680 TI - Simultaneous high-performance capillary electrophoresis analysis of the reduced and oxidised forms of ascorbate and glutathione. AB - We describe here a procedure for the simultaneous analysis of the oxidised and reduced forms of the major cellular hydrophillic antioxidants, ascorbic acid (vitamin C) and glutathione (gamma-L-glutamyl-L-cysteinylglycine), by high performance capillary electrophoresis. Separations are performed in uncoated fused-silica capillaries using 200 mmol/l borate pH 9.0, containing 20% (v/v) acetonitrile as the background electrolyte with fixed-wavelength UV absorbance detection at 185 nm. The influence of pH, organic solvent and other additives on the resolution of these compounds is described and we show that the optimised protocol is capable of simultaneously resolving other thiol components including, N-acetylcysteine and methyl-S-glutathione. The method is suitable for the analysis of these antioxidants in Arabidopsis and Nicotiana leaf tissue and is compatible with the use of the high ionic strength, acidic extraction solvents which are necessary to quench the redox equilibria of these labile components. PMID- 9342681 TI - High-performance capillary electrophoretic analysis of hyaluronan in effusions from human malignant mesothelioma. AB - A procedure to quantify hyaluronan in effusions from human malignant mesothelioma using a highly sensitive and reproducible high-performance capillary electrophoresis (HPCE) method is presented. Following ethanol precipitation, hyaluronan and galactosaminoglycans were degraded to delta 4,5-disaccharides with a mixture of chondroitinases ABC and AC. Heparan sulphate and proteins/glycoproteins were separated by ultrafiltration on a Centricon 3 membrane, and hyaluronan-derived disaccharides were analysed by direct injection of the filtrate into a HPCE system. Determination of hyaluronan in effusions from five healthy individuals and three patients with mesothelioma gave values comparable to those found using the HPLC method. One of the advantages of the HPCE method as compared to HPLC is the low solvent consumption. The much lower detection limit (attomole level) of the HPCE method may also allow the analysis of hyaluronan content in serum. The contribution of HPCE in diagnosis of a neoplasm, such as human malignant mesothelioma, illustrates the great potential of this technique in the field of life sciences. PMID- 9342683 TI - Paced Auditory Serial Addition Test: adult norms and moderator variables. AB - This study examined the performance of a sample of 821 healthy job applicants on the Paced Auditory Serial Addition Test (PASAT). Subjects had previously passed basic academic skills tests and physical examinations and were deemed free of cognitive impairment and medical illness. They were also motivated to perform well on cognitive tests. Gender, ethnicity, and education were not significant moderator variables in our subjects. Age and IQ did significantly affect PASAT test results. Normative data are stratified by age and WAIS-R Full Scale IQ scores to be useful to those who administer the PASAT in clinical practice. PMID- 9342682 TI - Determination of p-aminosalicylic acid and its N-acetylated metabolite in human urine by capillary zone electrophoresis as a measure of in vivo N acetyltransferase 1 activity. AB - A capillary zone electrophoresis method has been developed for the determination of p-aminosalicylic acid (PAS) and its metabolite, N-acetyl-p-aminosalicylic acid (N-acetyl-PAS), in urine. A linear relationship was observed between time normalized peak area and the concentration of the parent and metabolite with correlation coefficients greater than 0.9990. The method could be applied to the determination of PAS and N-acetyl-PAS in human urine without any sample pretreatment. A good separation of the analytes is achieved in a run time of 12 min (15 min total, including capillary wash). Using PAS as a probe for N acetyltransferase 1 activity, 20 healthy volunteers were phenotyped after oral administration of a 1 g dose. The preliminary results seem to indicate a bimodal distribution of N-acetyl-PAS/PAS molar ratios. PMID- 9342684 TI - A partial cross-validation of a Halstead-Reitan Battery malingering formula. AB - Using the Halstead-Reitan Battery profiles of 796 people, a formula for the detection of malingering was partially cross-validated to assess the false positive rate. Subjects included normals, psychiatric cases, and persons with all major types of brain disorder. The formula incorrectly designated 32% of the sample as fakers (i.e., as false positives). Of the 120 head-trauma cases, 39 (32%) obtained Fake scores, whereas 81 (67%) were correctly assessed as not malingering. The correlation of the results of the formula and the severity of the profile (as measured by the Average Impairment Rating) was 67, p < .0001. PMID- 9342685 TI - Criterion-related validity of the Geriatric Depression Scale in Alzheimer's disease. AB - The effect of cognitive impairment on the criterion-related validity of the self report Geriatric Depression Scale (GDS) was investigated in community-dwelling older adults. The GDS was administered to subjects with Alzheimer's disease (AD; n = 715) and normal cognitive function (NC; n = 93). The criteria consisted of the diagnosis of major depressive disorder and the total score on Hamilton Rating Scale for Depression, each derived from structured interview with a collateral informant to circumvent the effect of cognitive impairment on criterion measurement. Agreement between the GDS and both criteria was substantially attenuated in AD relative to NC subjects, differences primarily reflecting a lower than expected rate of item endorsement on the GDS in AD. A decrement in GDS validity coefficients was measurable across a broad range of cognitive impairment, suggesting limited utility of the scale in AD. PMID- 9342686 TI - Memory performance after head injury: contributions of malingering, litigation status, psychological factors, and medication use. AB - Impaired memory test performance can reflect a host of factors, such as head injury/postconcussive syndrome, involvement in litigation, malingering behavior, psychological distress, and medication use. Such factors are important in interpreting memory test performances in patients referred in the context of litigation. We examined memory test performance in mild head-injured patients in litigation, mild to moderate head-injured patients not in litigation, severely head-injured patients not in litigation, depressed patients, and patients with somatization disorders. Findings showed that several memory tests were useful in distinguishing probable malingerers from the other groups. There was a complex interaction among malingering status, psychological status, and medication use in the prediction of memory test results. Although nonneurological factors were related to memory impairment, litigation status alone was not predictive of memory performance. The results emphasize the need to consider nonneurological factors in the interpretation of poor memory performance in patients seen for forensic evaluation. PMID- 9342687 TI - Chronic obstructive pulmonary disease: effects of hypoxia on neurological and neuropsychological measures. AB - Eighteen patients with chronic obstructive pulmonary disease (COPD) were administered a series of pulmonary, neurological, and neuropsychological measures to test if there was an effect of COPD on neurological and cognitive functioning. Overall, there was no evidence of general dementia in this sample. Measures of immediate and delayed memory, complex attention, and speed of information processing correlated highly with arterial carbon dioxide partial pressure and, to a lesser extent, with oxygen partial pressure. Measures of language abilities, perceptual-motor functioning, and simple attention generally were not related to arterial gas pressures. A similar pattern of findings was obtained when group differences were examined between participants classified as severely hypoxic or mildly hypoxic, although group differences were mitigated by premorbid IQ differences. Hypoxia in COPD results in a relatively focused pattern of impairment in measures of memory function and tasks requiring attention allocation. The memory dysfunction may be related to involvement of limbic memory regions necessary for explicit memory. The attentional deficits were attributed to diffuse brain involvement resulting in reduced resource allocation. Early diagnosis and treatment of the hypoxia is essential. PMID- 9342688 TI - Attention deficits in stroke patients with aphasia. AB - Attentional capacity and sustained attention were investigated in 21 aphasic stroke patients and 21 non-brain-damaged patients. Attentional capacity was assessed using a series of reaction time (RT) tasks. The aphasic patients demonstrated impaired attentional capacity as shown by slower processing speed than the non-brain-damaged group (p < .01) and greater increases in RT with increased processing load (p < .05). Similar patterns were found for both verbal and spatial material. There was no significant relationship between severity of auditory comprehension deficits and attentional capacity. Sustained attention was assessed using a cognitive vigilance task requiring identification of a target letter presented infrequently over 32 minutes. Both the aphasic and the non-brain damaged group demonstrated a decline in performance with time on task as shown by a steady increase in RTs (p < .0001), but the decline was equivalent across the groups. Thus, the aphasic group did not show a specific deficit in the ability to sustain attention. PMID- 9342689 TI - Assessment of cognitive deterioration in individual patients following cardiac surgery: correcting for measurement error and practice effects. AB - Assessment of cognitive change in individual patients may be confounded by unreliability of test scores and effects of repeated testing. An index correcting for both problems is proposed and compared with change indices that do not or do not adequately deal with measurement error and practice effects. These indices were used to examine cognitive deterioration in a sample of 63 patients undergoing cardiac surgery. It was demonstrated that for test measures with a low reliability, failure to correct for measurement error resulted in overestimation of deterioration rates. For test measures with a high reliability, but showing substantial practice effects, failure to correct for practice effects resulted in underestimation of deterioration rates. With the proposed index, cognitive deterioration shortly after cardiac surgery was most frequently observed for attention and psychomotor speed, less frequently for verbal fluency, and only occasionally for learning and memory. PMID- 9342691 TI - A preliminary profile of neuropsychological deficits associated with major depression. AB - A profile of neuropsychological deficits of clinically depressed (major depression) but otherwise unimpaired individuals is presented, based on a meta analysis of all studies published since 1975 and meeting stringent methodological and sample selection criteria. Deficits are discussed separately for different cognitive areas in terms of mean size of deficit, variability between studies, variability of individual scores in depressed populations relative to that of controls, and expected proportion of depressed individuals scoring two standard deviations or more below the mean of controls. The neuropsychological deficits of individuals with major depression are shown to be consistent with a global diffuse impairment of brain functions with particular involvement of the frontal lobes. Recent neuro-imaging studies also indicating frontal dysfunction in clinical (functional) depression are referred to. Both the severity and the profile of cognitive deficiencies in depression are postulated to be similar to those seen in moderately severe traumatic brain injury. PMID- 9342692 TI - Massive and persistent anterograde amnesia in the absence of detectable brain damage: anterograde psychogenic amnesia or gross reduction in sustained effort? AB - The case of a young patient with severe and persistent anterograde amnesia of no known cause is reported. Anterograde amnesia arose within a 1-month period and has persisted for more than 1 year. Although a wide variety of neurological and neuroradiological assessments were completed (EEG, evoked potential recordings, Doppler sonography, MRI, PET), no evidence of brain damage was detected. Neuropsychologically, the patient was of high intelligence, had average to above average short-term memory, and normal retrograde memory abilities, but severe and persistent anterograde amnesia in both verbal and nonverbal domains. Furthermore, he demonstrated grossly reduced long-term concentration. It is likely that a complex chain of interacting variables can produce a syndrome that appears phenomenologically as anterograde amnesia without organically measurable correlates. PMID- 9342693 TI - Neuropsychology with Spanish speakers: language use and proficiency issues for test development. AB - The practice of neuropsychological assessment with Spanish speakers is hampered by language inadequacy in test translations and adaptations. The authors advance some ideas on how such testing materials have made their way into our field and suggest some ways to minimize such problems in the future. PMID- 9342690 TI - Neuropsychology of chronic fatigue syndrome: a critical review. AB - This article provides a comprehensive and critical review of the neuropsychological and related literature on chronic fatigue syndrome (CFS). Despite the methodological limitations observed in several studies, some consistent findings are noted. The most consistently documented neuropsychological impairments are in the areas of complex information processing speed and efficiency. General intellectual abilities and higher order cognitive skills are intact. Emotional factors influence subjective report of cognitive difficulty, whereas their effect on objective performance remains uncertain. Although the neuropathological processes underlying cognitive dysfunction in CFS are not yet known, preliminary evidence suggests the involvement of cerebral white matter. Directions for future research are outlined. PMID- 9342694 TI - Measurement of erythrocyte volumes in splenectomized horses and sham-operated horses at rest and during maximal exercise. AB - Erythrocyte volumes of thoroughbred horses were measured. The volumes of splenectomized horses and sham-operated horses 2 hr after injection of 50Cr tagged erythrocytes (at rest) and during maximal exercise were measured using the non-radioactive isotope 50Cr. Because splenic erythrocytes are released into circulation during exercise, it was estimated that the erythrocyte volumes of the sham-operated horses during maximal exercise are larger than those of the horses at rest. However, the erythrocyte volumes of the sham-operated horses at rest were about equal to those during maximal exercise. In the splenectomized horses, furthermore, erythrocyte volumes at rest and those at exercise were nearly equal. From these results, blood stored in the equine spleen is gradually mixed with circulating blood, and it was clarified that the phenomenon was completed within 2 hr. Although it is basically impossible to measure the circulating erythrocyte volume at rest using the erythrocyte tagged method, we observed that it is possible to measure the total erythrocyte volume using the 50Cr method. Also, the plasma volumes of the splenectomized horses during maximal exercise were found to be slightly smaller than those at rest. On the other hand, in the sham-operated horses, the plasma was decreased by a large quantity after maximal exercise. Therefore, it was suggested that the spleen participates in the phenomenon involving the disappearance of plasma from circulation due to exercise. PMID- 9342696 TI - The use of functional indexes to evaluate fitness in Andalusian horses. AB - The fitness of 8 Andalusian horses between 3 and 4 years of age was analysed. The animals were subjected to an exercise test on a sandy track consisting of 2 stages of different intensities. The first stage was of submaximal intensity at 4 speeds which increased progressively (4.17, 5.56, 6.94 and 8.33 m/sec.) covering a distance of 1,000 m in each level. Between each of these speeds, the horses rested for 2 min. The second stage was a maximal speed test over the same distance carried out 2 min after the ending of the maximal phase. Data of heart rate, plasma lactate concentration, velocity, PCV and pH in the blood were obtained. Maximum heart rate, maximum velocity, VLA2, VLA4, peak lactate, minimum pH and maximum PCV were considered functional indexes. A principal component enabled us to segregate horses according to their fitness and in relation to the information provided by the trainers in charge of these horses. The most discriminant variables in order to segregate horses were pHmin, VLA4, HRmax, VLA2 and Vmax. Differences between horses in relation to PCVmax were not found. The influence of each one of these functional indexes on the test exercise tolerance was discussed. PMID- 9342695 TI - Alteration of T-cell subsets in the lymph nodes from cats infected with feline immunodeficiency virus. AB - Alterations of T-cell subsets in the lymph nodes from FIV-infected cats in various clinical disease stages were examined histologically. In the early stage of infection (AP stage), follicular hyperplasia accompanied by expansion of the paracortical area was observed. Follicular involution and depletion with reduced paracortical area was observed in the ARC and AIDS stage nodes. The maximum section area of the entire popliteal lymph node was expanded significantly in the AP nodes. The paracortical area expanded in the AP nodes and decreased in the ARC and AIDS stage nodes. The cell density in the paracortical area in the AP nodes did not show a significant increase, while there was a significant reduction in the ARC and AIDS stage nodes. The lymph node CD4/CD8 ratio in the AP and ARC stages significantly decreased as compared with that of uninfected control cats, but conversion of the ratio was not seen. The estimated total numbers of CD4+ and CD8+ cells in the maximum section were increased in the AP stage but significantly decreased in the ARC and AIDS stages. Our study indicated that the lymphocyte depletion in the terminal ARC and AIDS stages of FIV infection was associated with both CD4+ cells and CD8+ cells. Findings obtained in this study might provide useful information for studying the pathophysiology of FIV infection. PMID- 9342697 TI - Suppression of glucose absorption by extracts from the leaves of Gymnema inodorum. AB - Gymnema sylvestre (GS) is one of the Asclepiad strains that grows in South-east Asia. Their therapeutic effects for treating diabetes mellitus, rheumatic arthritis and gout have been well known for a long time. However, the problem is that GS suppresses sweetness and tastes bitter. For this study, we chose Gymnema inodorum (GI) instead of GS, since it has an advantage that it does not suppress sweetness nor is it bitter in taste. In this paper, effects of glucose availability of some saponin fractions (F-I to F-IV) extracted from GI leaves, which were obtained by high-performance liquid chromatography were studied on a high K(+)-induced contraction of guinea-pig intestinal smooth muscle, O2 consumption on guinea-pig ileum, glucose-evoked transmural potential difference (delta PD) of guinea-pig everted intestine and blood glucose level in glucose tolerance tests on rats. The extracts of GI leaves suppressed the intestinal smooth muscle contraction, decreased the O2 consumption, inhibited the glucose evoked-transmural potential, and prevented the blood glucose level. Our studies suggest that the component of GI inhibits the increase in the blood glucose level by interfering with the intestinal glucose absorption process. PMID- 9342698 TI - Radioimmunoassay of saliva estrone sulfate in pregnant sows. AB - The aim of this study was to establish radioimmunoassay (RIA) for saliva estrone sulfate (E1S), and to elucidate changes in saliva E1S during pregnancy in the sow. Saliva E1S was extracted using a commercially available solid phase column, and the E1S fraction obtained was subjected to RIA. The sensitivity of the RIA was 29.7 pg/tube. The intra- and inter-assay coefficients of variation were 5.5 8.4% and 13.1-19.5%, respectively. Mean recovery for E1S added to saliva samples was as high as 99.9%. A significant positive correlation (r = 0.54, n = 69, p < 0.01) existed between saliva and plasma E1S concentrations. During gestation, the changing patterns of saliva and plasma E1S concentrations were essentially the same, and two peaks of E1S concentrations were observed, one around day 30 and another just before parturition, although E1S concentrations in saliva remained at only 2.4-38.1% (mean 11.4%) of those in plasma E1S. Thus, the present study has made it possible to measure saliva concentrations of E1S and demonstrated a high degree of positive correlation between saliva and plasma E1S concentrations. These results suggest that diagnosis of early pregnancy and of normal or abnormal fetal development could be made by measurements of E1S in saliva. PMID- 9342699 TI - Disposition of ampicillin in honeybees and hives. AB - Disposition profile of ampicillin (ABPC) among honeybees, larvae, honey and royal jelly in a hive after oral dosing to adult bees was studied. Four honeybee colonies were administered the single dose of ABPC at the rate of 30 mg/hive by addition to sugar syrup or pollen substitute (paste) for 1 day intake. The colonies received ABPC in syrup showed high drug residue levels in honey and it lasted over 14 days beyond the detection limit of residual analysis. In the hives given ABPC in paste, relatively low honey residues were found, however, the distributions of the drug in young larvae and jelly which was the food of the larvae were very low. ABPC was considered to be a promising drug for the control of American foulbrood, an important bacterial disease of honeybee larvae, because of its high antibacterial activity to the pathogen, Paenibacillus larvae, and instability of residue in honey as human food. The low distribution in young larvae, the target of the disease, threw a doubt on the efficacy of ABPC for American foulbrood control. PMID- 9342700 TI - Abnormal G1 arrest in the cell lines from LEC strain rats after X-irradiation. AB - The effect of X-irradiation of cell lines from LEC and WKAH strain rats on a progression of cell cycle was investigated. When WKAH rat cells were exposed to 5 Gy of X-rays and their cell cycle distribution was determined by a flow cytometer, the proportion of S-phase cells decreased and that of G2/M-phase cells increased at 8 hr post-irradiation. At 18 and 24 hr post-irradiation, approximately 80% of the cells appeared in the G1 phase. On the contrary, the proportion of S-phase cells increased and that G1-phase cells decreased in LEC rats during 8-24 hr post-irradiation, compared with that at 0 hr post irradiation. Thus, radiation-induced delay in the progression from the G1 phase to S phase (G1 arrest) was observed in WKAH rat cells but not in LEC rat cells. In the case of WKAH rat cells, the intensities of the bands of p53 protein increased at 1 and 2 hr after X-irradiation at 5 Gy, compared with those of unirradiated cells and at 0 hr post-irradiation. In contrast, the intensities of the bands were faint and did not significantly increase in LEC rat cells during 0 6 hr incubation after X-irradiation. Present results suggested that the radioresistant DNA synthesis in LEC rat cells is thought to be due to the abnormal G1 arrest following X-irradiation. PMID- 9342701 TI - Phylogenetic analyses of Staphylococcus based on the 16S rDNA sequence and assignment of clinical isolates from animals. AB - The nucleotide sequences of the 16S rDNA in 17 strains of 16 taxa of the genus Staphylococcus were determined. The sequences were compared phylogenetically together with the gene sequences of 10 (including 7 other species) Staphylococcus species retrieved from the DNA Data Bank of Japan. Although the primary and secondary structures of most of Staphylococcus species were very similar (homology values 96.4% or more) except for S. caseolyticus MAFF 911387T (homology values 95.4% or less), the 23 staphylococcal species were divided into 10 groups based on similarity, evolutionary distance and phylogenetic tree analysis. Nucleotide stretches in several variable domains in the 16S rDNA sequences appeared to be specific for the bacterial groups or the species. By comparing such characteristics in the sequence and phylogenies of 5 staphylococcal clinical isolates from bovine mastitis, canine and feline pyoderma, and feline urogenital syndrome with the information obtained in this study, the species level of each organism was identified. PMID- 9342702 TI - Apoptosis of murine hepatocytes induced by high doses of galactosamine. AB - Apoptosis induced by high doses of Galactosamine (GalN) was investigated in mice hepatocytes in vivo. In mice intraperitoneally (i.p.) treated with GalN 3 g/kg, the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) positive cells were first observed at 6 hr postadministration (PA). Both acidophilic bodies in hematoxylin and eosin (HE)-stained sections and TUNEL positive cells were markedly found at 24 hr PA. At 48 hr PA, cellular degeneration and necrosis of hepatocytes were prominently observed, and TUNEL positive cells were scarcely found. In the mice ip treated with GalN 1.5 g/kg, the lesion was milder than that in those treated with GalN 3 g/kg. Acidophilic bodies and TUNEL-positive cells were scarcely found at 24 hr PA, whereas they were markedly seen at 48 hr PA. In addition, a ladder-like DNA fragmentation pattern by agarose gel electrophoresis was observed most remarkably at 24 hr PA with GalN 3 g/kg and at 48 hr PA with GalN 1.5 g/kg, and less distinctly at 48 hr PA with GalN 3 g/kg. On the other hand, sGOT and sGPT activities increased prominently at 48 hr PA with GalN 3 g/kg. These results suggest that the cell death induced by high dose of GalN may be caused by apoptosis, and subsequently by necrosis in vivo. PMID- 9342703 TI - Postpartum plasma PGF metabolite profile in cows with dystocia and/or retained placenta, and effect of fenprostalene on uterine involution and reproductive performance. AB - Objectives of this study were to show postpartum plasma PGF2 alpha metabolite (PGFM) profile, to clarify whether endogenous PGF2 alpha plays a certain role in the uterine involution in cows with dystocia and/or retained placenta, and to examine the effects of fenprostalene, a long-acting PGF2 alpha analog, on the uterine involution and reproductive performance of the cows with abnormal puerperium. A group of 27 cows with dystocia and/or retained placenta showed a massive release of PGF2 alpha after parturition as indicated by a rise of plasma concentrations of PGFM, significantly higher than 33 cows with normal puerperium. The duration of the elevated plasma PGFM concentrations in the cows with abnormal puerperium was shorter than that of the normal cows. In cows with normal puerperium, those showing relatively longer duration of elevated plasma PGFM levels needed a shorter period for postpartum uterine involution than the cows showing a shorter duration of the PGFM elevation (P < 0.01), while no such relationship was observed in cows with abnormal puerperium. In field trials, an administration of an exogenous PGF2 alpha, fenprostalene, at 7 to 10 days (78 cows) or 14 to 28 days postpartum (74 cows) was found to be effective in facilitating uterine involution and resumption of ovarian cyclicity, and improved reproductive performance. It may be concluded that a large amount of PGF2 alpha is released for a relatively shorter period in cows after dystocia and/or retained placenta and the elevation of PGFM is not responsible for the uterine involution. The administration of the exogenous PGF2 alpha was shown to be effective at improving the postpartum reproductive performance of cows with abnormal puerperium. PMID- 9342704 TI - Antigenic differentiation of turkey rhinotracheitis virus strains using monoclonal antibodies and polyclonal antisera. AB - Monoclonal antibodies (mAbs) were prepared against the 8597/CV94 strain of turkey rhinotracheitis virus (TRTV). These mAbs were used to investigate antigenic relationships among three strains (8597/CV94, 1162/92 and CVL14/1 strain) of TRTV, together with polyclonal chicken and rabbit antisera to 8597/CV94 strain, and guinea pig antisera to each of the three strains. Thirty mAbs to the glycoprotein (G:3 clones), fusion (F1:6 clones), phosphorylated (P:6 clones), nucleocapsid (N:12 clones), and matrix (M:3 clones) proteins of viral antigen were obtained by cell fusion. Among these, two mAbs to F1 protein showed virus neutralizing activity. The results of ELISA test indicated that some mAbs only reacted to the 8597/CV94 strain, some reacted to 8597/CV94 and 1162/92 strains, and others reacted to all three viral strains. In neutralization tests with the three virus strains, polyclonal chicken and rabbit antisera against the 8597/ CV94 strain showed the same antibody titers. Results with four neutralizing mAbs including two previously reported mAbs [Ref. 21] indicated the titers of two mAbs (Pn2-2E and Pn3-2F) to 8597/CV94 were much higher than those to the other two viral strains. No differences were observed in the titers of the other two mAbs (Pn01-8E and Pn06-4D) against any viral strains. In cross-neutralization tests with polyclonal guinea pig antisera, there was some variations among viral strains. This work demonstrated that the Japanese isolate 8597/CV94 of TRTV is somewhat different in antigenicity from two British isolates from chickens and turkeys. PMID- 9342706 TI - Computed tomography and magnetic resonance findings in two dogs and a cat with intracranial lesions. AB - Two dogs and a cat with intracranial lesions were evaluated by both computed tomography (CT) and magnetic resonance (MR) imaging. In a dog with vestibular syndrome, better quality images of the medulla oblongata surrounded by thick bones were obtained by MR than by CT, on which the appearance of artifacts impeded the clear image of the area. In a dog with multiple brain metastases of lymphoma, contrast CT delineated lesions more clearly than MR, which was performed one week after CT. During that week dexamethasone which might affect the clarity of MR images of the lesion was administered to reduce brain edema. In a cat with meningeal syndrome of lymphocytic leukemia, only contrast MR imaging identified the width and site of the lesion. These results indicate that it is necessary to select either one of these imaging methods according to the type and site of lesions that are suspected in a particular case. PMID- 9342705 TI - Cardiovascular reflex mechanisms by topical instillation of capsaicin and distilled water into the larynx in anesthetized dogs. AB - Cardiovascular reflex mechanisms by topical laryngeal instillation of capsaicin (CAPS) or distilled water were evaluated in anesthetized chronic tracheostomized dogs. Both CAPS (10 micrograms/ml) and water instillation into the isolated upper airway caused a significant decrease in heart rate (P < 0.05) and a significant increase in blood pressure (P < 0.05) from the values before instillation under both spontaneous and controlled ventilation. The bradycardia was significantly reduced by atropine pretreatment (P < 0.05) and the hypertension was significantly decreased by phentolamine and propranolol pretreatments (P < 0.01). A higher concentration of CAPS (100 micrograms/ml) instillation considerably reduced the response to subsequent CAPS (100 micrograms/ml) instillation, whereas the response to water was sustained, indicating the desensitization of laryngeal CAPS-sensitive endings. All the reflex responses to CAPS and water were eliminated by topical anesthesia with lidocaine. It was concluded that the laryngeal cardiovascular reflex responses were mediated by the afferents such as the laryngeal CAPS-sensitive presumably C-fiber endings or water-responsive receptors and by both the parasympathetic and sympathetic nervous systems as efferents. PMID- 9342707 TI - Effect of simultaneous bilateral tibial nerve stimulation on somatosensory evoked potentials (SEP) in dogs. AB - Since somatosensory evoked potentials (SEP) by hindlimb nerve stimulation are known to ascend bilaterally in the spinal cord, it was investigated whether or not simultaneous bilateral stimulation causes facilitation or inhibition of the stimuli. In an experiment using 36 adult Beagle dogs, the difference between simultaneous bilateral stimulation and unilateral stimulation was studied as to latencies and amplitudes. No significant difference was noted. However, the detection of far-field potentials by bilateral stimulation was not effective in which case far-field potentials did not recorded by unilateral stimulation. It was therefore confirmed that simultaneous bilateral stimulation does not cause facilitation or inhibition in the relay pathways, and does not affect the latency. PMID- 9342708 TI - Apoptosis of enterocytes induced by inoculation of a strain of attaching and effacing Escherichia coli and verotoxin. AB - When verotoxin (VT)-producing attaching and effacing Escherichia coli (AEEC, serotype O5: H-) were inoculated perorally into 10-day-old rabbits, attaching of the E. coli to enterocytes and effecting of their microvillous portion were observed extensively from the ileum to the colon. Subsequent apoptotic changes of the infected enterocytes were demonstrated by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) reaction and electron microscopy. Apoptosis was also induced in cultured Vero cells by inoculation of VT extracted from the AEEC. This study clarified that VT-producing AEEC induce apoptosis of enterocytes, causing mucosal damage. PMID- 9342709 TI - Molecular cloning of a canine metallothionein cDNA. AB - A canine metallothionein cDNA obtained from the liver of a cadmium-treated beagle was cloned and sequenced. Asn at position 4 conserved among all mammalian metallothionein-1 and metallothionein-2 is replaced by Asp in the canine metallothionein cDNA clone. Because the acidic amino acid doesn't exist at either position 10 or 11 in the deduced amino acid sequence, it is supposed that this cDNA is derived from canine metallothionein-1 mRNA. Northern blot analysis using the cDNA as a probe revealed the induction of the canine metallothionein mRNA expression in the liver and kidney of a cadmium-treated beagle. Thus, the canine metallothionein cDNA obtained in the present study should provide an useful tool for the molecular investigation of metallothionein in dog. PMID- 9342710 TI - An improved canine model of subarachnoid hemorrhage using intrathecal indwelling catheters. AB - In the present study, the feasibility of intrathecal indwelling catheters in the preparation of a repeated subarachnoid hemorrhage (SAH) model in dogs, as well as chronic intrathecal administration of therapeutic agents against the ensuing cerebral vasospasm was examined. Briefly, through a small suboccipital incision, two catheters were introduced into the subarachnoid space so that their tips were positioned in the prepontine cistern. One was used to induce SAH by infusing autologous blood, and the other to administer pharmacological agents (saline and/or saline containing a dye in this study) by means of an osmotic pump. The occurrence of cerebral vasospasm was followed by angiography via the catheter placed in the vertebral artery. The obtained results show: i) the injected blood effectively formed a subarachnoid clot in the prepontine cistern, invariably leading to the occurrence of severe cerebral vasospasm of the basilar artery; ii) the fluid injected by the osmotic pump was evenly distributed in the cisterns around the brain stem; iii) on post mortem pathological examination, no injury of the brain or the major arteries ascribable to the placement of catheters was found. Therefore, the present model is considered to be useful for both the investigation of pathophysiology and therapy of cerebral vasospasm following SAH, to be more favorable from the standpoint of animal protection, and more convenient and reliable than those used until now. PMID- 9342711 TI - A polymorphism observed in the experimentally successful peptide vaccine sequence derived from Theileria sergenti piroplasm major surface antigen (p33). AB - A polymorphism in the experimentally successful peptide vaccine sequence (EVVWKEKKEVKDLDA, amino acids 134-148) derived from the 33 kDa piroplasm major surface antigen (p33) of Theileria sergenti was examined. The vaccine sequences obtained by PCR amplification and sequencing of the p33 gene from a total of 15 parasite-infected cattle blood samples collected from 4 prefectures through Hokkaido to Kumamoto revealed the two major sequences (Ikeda and Chitose stock types) either of which was identified in all samples. Since the peptide vaccine develops the parasite species- or stock-specific immunity in the animals, an application of the two major peptide sequences as cocktailed vaccine should be evaluated for a practical use of this strategy to controlling T. sergenti infection in Japan. PMID- 9342712 TI - Gangliocytoma with immature neuronal cell elements in the pituitary of a rat. AB - A spontaneous pituitary gangliocytoma with abundant, immature neuronal cell elements was found incidentally in a 109-week-old female Fischer 344 rat. The pituitary parenchyma was largely occupied by a tumor nodule with necrotic and hemorrhagic foci and cyst. The tumor was composed of mature ganglion-like (M) cells, small immature ganglion (I) cells and transitional (T) cells, with a fibrillar matrix. The I and T cells were intermingled with the M cells or were arranged in compact clusters, in which the I cells formed perivascular rosette like structures, sometimes with mitotic figures. Immunohistochemically, all types of tumor cells were positive for neuron-specific enolase, and only the M cells was positive for chromogranin A. This result may be correlated with the degree of cytodifferentiation. PMID- 9342713 TI - Restriction fragment length polymorphism analysis of Akabane virus nucleoprotein gene. AB - The nucleoprotein genes of Akabane virus S RNA segment from 21 Japanese and two Australian isolates were amplified by the reverse transcriptase-polymerase chain reaction (RT-PCR) using a primer set containing the initiation and termination codon of the gene. The RT-PCR products were sufficiently produced from the purified virion RNAs of all the isolates, and then analyzed by enzymatic digestion with 11 restriction endonucleases. Digestion with the eight restriction enzymes revealed sequence variation of the isolates. Restriction fragment length polymorphism (RFLP) profiles obtained by digestion revealed the existence of four major groups (genogroups) among the isolates. The two Australian isolates had extremely different RFLP profiles than the Japanese isolates. The data demonstrate the usefulness of analyzing the RFLP patterns to understand the genetic variability of AKA virus isolated in Japan and Australia. PMID- 9342714 TI - Quantitative measurement of antibody inhibiting reverse transcriptase activity in cats naturally infected with feline immunodeficiency virus. AB - Quantitative measurement of reverse transcriptase-inhibiting (RTI) antibodies in Japanese household cats naturally infected with feline immunodeficiency virus (FIV) was performed by poly A-linked colorimetric reverse transcriptase assay (PAC-RTA). Eight FIV-seropositive plasma samples were diluted twofold from 1:10 to 1:160 and incubated with FIV RT. Fifty percent RTI activity (RTI50) was calculated from a dose response PAC-RTA curve. The plasma of FIV-seropositive cats showed different RTI activities against two Japanese isolates and Petaluma strain. Six of eight plasma samples showed RTI activities against the Japanese isolates (subtype B), but only one showed RTI activity against Petaluma strain (subtype A). It is important to use the appropriate strain as a source of RT for detection of RTI antibody in cats. PMID- 9342715 TI - Successful laparoscopy assisted ovariohysterectomy in two dogs with pyometra. AB - Two dogs with pyometra were treated by laparoscopy assisted ovariohysterectomy. Hemostasis of the mesovarium was achieved with an ultrasonic scalpel and hemoclips. Both ovaries and the uterus were exposed via a 10-mm caudal port that was enlarged to 3 cm and the uterine cervix was excised after ligation of the uterine arteries. These cases were the first report on ovariohysterectomy for pyometra by laparoscopy assisted surgery in the veterinary field. PMID- 9342716 TI - Two-step PCR in the evaluation of antibiotic treatment for Ehrlichia platys infection. AB - We evaluated the feasibility of using the two-step polymerase chain reaction (PCR) in determining the withdrawal time of antibiotic treatment for Ehrlichia platys infection. We also present experimental evidence of a dog remaining a carrier after treatment with tetracycline. Canine infectious cyclic thrombocytopenia (CICT) was induced in 3 dogs by intravenous inoculation of blood infected with E. platys. Tetracycline was administered to one of the dogs for 2 weeks when parasitemia appeared. Although the hematologic abnormality of cyclic thrombocytopenia soon disappeared, a few parasitized platelets reappeared after the withdrawal of treatment, and the dog thus remained as a carrier. The other dogs were treated with doxycycline when parasitemic episodes first developed. The durations of antibiotic regimens were determined by the results of two-step PCR in which the 16S rDNA of E. platys was amplified from blood samples. Doxycycline was withdrawn after 8 days of treatment, and the follow-up monitoring continued for 3 weeks. The platelet counts of the 2 dogs remained within the normal range, and the etiologic agent of CICT was not found either by Giemsa staining or by the two-step PCR, indicating complete elimination of the agent. PMID- 9342717 TI - Detection of Clostridium septicum hemolysin gene by polymerase chain reaction. AB - A polymerase chain reaction (PCR) was developed for the detection of the hemolysin (alpha toxin) gene of Clostridium septicum. The PCR primers were designed from the sequence of the hemolysin gene and synthesized. A DNA fragment of 270 bp was amplified from 10 strains of C. septicum, but was not from strains of C. chauvoei, C. perfringens, C. novyi, or C. haemolyticum. When the PCR product was digested with Sau3AI, two DNA fragments of the expected 148 bp and 122 bp were recognized. The lowest detectable threshold of PCR for the hemolysin gene was 3.8 x 10(3) cells/ml. The PCR technique may be useful for rapid detection or identification of C. septicum associated with malignant edema. PMID- 9342718 TI - A staged protocol for the treatment of persistent pulmonary hypertension of the newborn. AB - The purpose of this report is to propose a staged therapeutic protocol for the treatment of persistent pulmonary hypertension of the newborn (PPHN) based on retrospective clinical experience. We analysed the clinical course of 19 term and near-term neonates with severe PPHN treated between 1991 and 1993, before the introduction of inhalational nitric oxide (NO) therapy. Basic therapy consisted of continuous sedation, and analgesia, circulatory support with dobutamine, dopamine and substitution of 5% albumin or packed red blood cells, mechanical hyperventilation, alkalisation with sodium bicarbonate and surfactant instillation. Consecutive therapy included the administration of norepinephrine, high frequency oscillatory ventilation (HFOV), Prostacyclin (PGI2) and extracorporeal membrane oxygenator (ECMO) therapy with entry criteria for each stage of treatment. From our observations we suggest that firstly, an early increase in systemic mean arterial pressure due to norepinephrine and secondly, HFOV trials are beneficial in patients with PPHN. The staged protocol shall be applicable in most neonatal intensive care units in which inhalational NO and ECMO therapies are not available and includes our present entry criteria for both therapies. PMID- 9342719 TI - Lloyd A. Horrocks: a great neurochemist of our time. PMID- 9342720 TI - Protein kinase inhibitors block neurite outgrowth from explants of goldfish retina. AB - A role for protein phosphorylation in the process of neurite outgrowth has been inferred from many studies of the effects of protein kinase inhibitors and activators on cultured neurotumor cells and primary neuronal cells from developing brain or ganglia. Here we re-examine this issue, using a culture system derived from a fully differentiated neuronal system undergoing axonal regeneration--the explanted goldfish retina following optic nerve crush. Of the relatively non-selective protein kinase inhibitors employed, H7, staurosporine and K252a were found to block neurite outgrowth, whereas HA1004 had no effect, a result which appears to rule out a critical role for protein kinase A. The more selective protein kinase C inhibitors, sphingosine, calphostin C and Ro-31-8220 were all inhibitory, as was prolonged treatment with phorbol ester and the protein phosphatase inhibitor okadaic acid. These results are in support of a role for protein kinase C in axonal regrowth. PMID- 9342721 TI - Ethanol-induced cell death by lipid peroxidation in PC12 cells. AB - Free radical generation is hypothesized to be the cause of alcohol-induced tissue injury. Using fluorescent cis-parinaric acid and TBARS, lipid peroxidation was shown to be increased in the presence of trace amounts of free ferrous ion in PC12 cells. This increase in lipid peroxidation was enhanced by ethanol in a dose dependent manner and also correlated with loss of cell viability, as measured by increased release of lactate dehydrogenase (LDH). Resveratrol, a potent antioxidant, had a protective effect against lipid peroxidation and cell death. These findings strongly suggest that ethanol-induced tissue injury and cell death is a free radical mediated process, and may be important in alcohol-related premature aging and other degenerative diseases. PMID- 9342722 TI - Effect of hypoxia and dopamine on arachidonic acid metabolism in superior cervical ganglion. AB - Superior cervical ganglion (SCG) may play a modulatory role on ventilatory control through its efferent sympathetic fibres, which innervate cells in the carotid bodies. In this study the in vivo effect of acute hypoxia versus normoxia on arachidonic acid (AA) metabolism was investigated in cat SCG. Using SCG homogenate AA was incorporated into glycerolipids of normoxic SCG in the following order: neutral glycerolipids > phosphatidylcholine (PtdCh) > phosphatidylinositol (PtdIns) > phosphatidylethanolamine (PtdE) > phosphatidylserine (PtdS) > and phosphatidic acid (PA). In vivo hypoxic treatment caused a significant decrease in incorporation of [1-14C]AA into PtdIns. Hypoxia had no significant effect on the level of AA radioactivity in diacylglycerol (DAG) as compared to control but significantly enhanced the level of arachidonoyl CoA (AA-CoA) radioactivity. It was observed that dopamine (DA) one of the most important neurotransmitter in SCG decreases AA uptake into phospholipids of normoxic SCG. In normoxic SCG, DA significantly decreased, AA incorporation into PtdCh, PtdIns and DAG. Moreover, DA decreased the level of AA-CoA radioactivity. Hypoxia and dopamine has no effect on AA metabolism in medulla oblongata isolated from the same animals. These results indicate that arachidonic acid metabolism in SCG is sensitive to hypoxia and dopamine action. Moreover, these results indicate that hypoxia inhibits selectively AA incorporation on the level of acylCoA lysophosphatidylinositol-acyltransferase. PMID- 9342723 TI - Regulation of phosphatidylethanolamine degradation by enzyme(s) of subcellular fractions from cerebral cortex. AB - Hydrolysis of 1-acyl-2-[14C]arachidonoyl-sn-glycero-3-phosphoethanolamine was studied in cerebral cortex homogenate and subcellular fractions. The enzyme(s) confined to the synaptic plasma membrane (SPM) hydrolyze(s) [14C arachidonoyl]phosphatidylethanolamine (PE) in the presence of EGTA to [14C arachidonoyl]diacylglycerol (DAG) and a small amount of [14C]arachidonic acid (AA). Degradation of PE is time-, protein- and substrate-dependent with a pH optimum of 7.8. The highest activity of PE degradation was observed in the presence of 10 mM EGTA. Under this condition GTP gamma S has no effect on PE hydrolysis. In the presence of Ca2+ ions degradation of PE was significantly lower as compared to the conditions with EGTA. However, the percentage distribution of free AA in the sum of both products of PE hydrolysis (AA + DAG) increases from 16 and 20% observed in the presence of EGTA 2 mM and 10 mM to 34% and 43% in the presence of 0.5 mM CaCl2 alone and together with GTP gamma S, respectively. Cytosolic enzymes also degrade PE in the presence of 2 mM EGTA with the formation of DAG and AA. Radioactivity in the AA represents about 80% of the total radioactivity of the products of PE degradation. The hydrolysis of PE by cytosolic enzymes is almost completely inhibited by neomycin but the hydrolysis by the SPM-bound enzyme(s) is inhibited only 70%. Other studies with quinacrine indicated that only a small pool of PE is degraded by SPM-bound Ca(2+) independent phospholipase A2 (PLA2). All of these data suggest that PE in cerebral cortex is mainly degraded by cytosolic and SPM-bound Ca(2+)-independent phospholipase C. Further studies towards a better understanding of the mechanisms of cerebral degradation and the physiological significance of Ca(2+)-independent pathways of PE hydrolysis are necessary. PMID- 9342724 TI - Effects of maturation on the phospholipid and phospholipid fatty acid compositions in primary rat cortical astrocyte cell cultures. AB - Phospholipid and phospholipid fatty acid compositional changes were studied in rat cortical astrocytes during dibutyryl cyclic adenosine monophosphate (dBcAMP, 0.25 mM) treatment starting after 14 days in culture (DIC). After 15 DIC, ethanolamine- and choline glycerophospholipid levels were increased 1.2- and 1.3 fold, respectively in treated compared to control cells. However, after 21 and 28 DIC, these levels were not significantly different between groups. Both groups had an increase in phosphatidylserine levels with increasing time in culture. Similarly, ethanolamine plasmalogen levels were transiently elevated after 21 DIC, but returned to previous levels after 28 DIC. The phospholipid fatty acid compositions for the acid stable and labile ethanolamine- and choline glycerophospholipids indicated that in dBcAMP treated cells, 20:4 n-6 and 22:6 n 3 proportions were elevated with increasing time in culture relative to control cells. As 20:4 n-6 proportions increased, there was a concomitant decrease in 20:3 n-9 proportions, suggesting an up regulation of n-6 series elongation and desaturation. In contrast, in control cells, the 20:4 n-6 proportions decreased with a corresponding increase in the 20:3 n-9 proportions. Thus, in treated cells, the cellular phospholipid fatty acid composition was dramatically different than control cells, suggesting that dBcAMP treatment may act to increase fatty acid elongation and desaturation. PMID- 9342726 TI - Synthesis of ethanolamine phosphoglycerides in rat cerebral cortex subjected in vitro to experimental hypoxia with and without hypocapnia. AB - Slices and homogenates from rat cerebral cortex were used to study the effect of hypoxia, with or without hypocapnia, on phosphatidylethanolamine synthesis. The incorporation of [1-3H]ethanolamine into the corresponding phospholipid was greatest in slices treated with pure nitrogen, intermediate when the nitrogen contained 5% CO2, and least in slices treated with 95% O2-5% CO2. The role of hypocapnia in reinforcing the effect due to hypoxia did not require the integrity of the cell because similar results were obtained by treating homogenates with pure nitrogen or nitrogen plus 5% CO2. In both cases the synthesis of phosphatidylethanolamine was abolished by the addition of EGTA and the degradation of newly synthesized phospholipid by phospholipases was similar to that obtained in controls. When the homogenate was not buffered, changes in the pH due to experimental treatment influenced the response to Ca2+ and to hypoxia plus hypocapnia. Intracellular calcium ions are thought to play a role in the response of cerebrocortical slices to N2-treatment. In fact, although the incorporation was greater in complete medium that contains 2 mM Ca2+ than in the same medium prepared without the addition of this ion, the relative increase of incorporation due to N2-treatment was greater in the medium lacking added Ca2+. PMID- 9342725 TI - Lipid-dependent modulation of Ca2+ availability in isolated mossy fiber nerve endings. AB - An enhancement of glutamate release from hippocampal neurons has been implicated in long-term potentiation, which is thought to be a cellular correlate of learning and memory. This phenomenom appears to be involved the activation of protein kinase C and lipid second messengers have been implicated in this process. The purpose of this study was to examine how lipid-derived second messengers, which are known to potentiate glutamate release, influence the accumulation of intraterminal free Ca2+, since exocytosis requires Ca2+ and a potentiation of Ca2+ accumulation may provide a molecular mechanism for enhancing glutamate release. The activation of protein kinase C with phorbol esters potentiates the depolarization-evoked release of glutamate from mossy fiber and other hippocampal nerve terminals. Here we show that the activation of protein kinase C also enhances evoked presynaptic Ca2+ accumulation and this effect is attenuated by the protein kinase C inhibitor staurosporine. In addition, the protein kinase C-dependent increase in evoked Ca2+ accumulation was reduced by inhibitors of phospholipase A2 and voltage-sensitive Ca2+ channels, as well as by a lipoxygenase product of arachidonic acid metabolism. That some of the effects of protein kinase C activation were mediated through phospholipase A2 was also indicated by the ability of staurosporine to reduce the Ca2+ accumulation induced by arachidonic acid or the phospholipase A2 activator melittin. Similarly, the synergistic facilitation of evoked Ca2+ accumulation induced by a combination of arachidonic acid and diacylglycerol analogs was attenuated by staurosporine. We suggest, therefore, that the protein kinase C-dependent potentiation of evoked glutamate release is reflected by increases in presynaptic Ca2+ and that the lipid second messengers play a central role in this enhancement of chemical transmission processes. PMID- 9342727 TI - The time course of malondialdehyde production following impact injury to rat spinal cord as measured by microdialysis and high pressure liquid chromatography. AB - This paper reports a highly sensitive, specific, and reproducible method for the analysis of malondialdehyde (MDA) from microdialysates. The microdialysates were reacted with 2-thiobarbituric acid, and the TBA adducts were separated by HPLC and detected using a fluorescence detector. Butylated hydroxytoluene was used as an antioxidant to minimize formation of artifacts. The time course of MDA production following impact injury to the rat spinal cord was obtained using this improved method. MDA concentrations in the extracellular space gradually increased from a basal level of 20 +/- 3.6 nM to 44 +/- 18.1 nM during the first 2 hr, reached a maximum of 95 +/- 19.8 nM at 5 hr, and then decreased to 36 +/- 9.5 nM at 9 hr. The findings support the hypothesis that spinal cord injury leads to increased membrane lipid peroxidation. PMID- 9342728 TI - Ion exclusion chromatography: parameters influencing retention. AB - Ion Exclusion Chromatography (IEC) finds application in the separation of a wide range of small, neutral or partially ionized molecules. In IEC, the strong as well as weak electrolytes are eluted unseparated, the first at the beginning and the latter at the end of the elution. The retention volumes of the remaining electrolytes are found to be proportional to their dissociation constant values. The dead and inner volumes of the chromatographic column can be determined from the observed dependence of retention volumes on dissociation constant values. The retention mechanism is described by the analytical equations and by the results obtained from the computer simulation of the column performance (using global thermodynamic and chromatographic equations or the Craig method). The mixed retention mechanism involving hydrophobic adsorption and screening effect is observed for weak electrolytes and aromatic compounds. Aromatic compounds are found to be retained almost solely by a reverse-phase mechanism involving interaction of the solute with the unfunctionalized regions of the stationary phase. The purpose of this paper is to survey the field. Using theoretical and experimental approaches, I show how different parameters can influence ion exclusion solute retention. Although this retention is affected by the physicochemical parameters of the sorbent, stationary and mobile phases especially, this study primarily deals with the structural solute parameters, stationary phase form and column temperature, that have had little or no discussion in literature. PMID- 9342729 TI - Elevation of platelet activating factor, inflammatory cytokines, and coagulation factors in the internal jugular vein of patients with subarachnoid hemorrhage. AB - The aim of the present study was to examine the changes of inflammatory and coagulation factors in blood of the internal jugular vein, not of peripheral vein, in patients with subarachnoid hemorrhage (SAH). The results show that while interleukin-6 (IL-6) and platelet activating factor (PAF) concentrations increased within first 4 days after SAH and remained elevated up to 14 days, interleukin-1 beta (IL-1 beta) showed a transient increase between 5-9 days after SAH and tumor necrosis factor-alpha (TNF-alpha) remained unchanged. Also different coagulation factors were increased between 5-9 days after SAH. Moreover, patients with delayed ischemic neurological deficits (DIND) displayed the highest levels of PAF and the coagulation factors, von Willebrand factor (vWF) and thrombin-antithrombin III complex (TAT). These results suggest that elevation of PAF and other inflammatory cytokines following SAH may cause the hypercoagulation state that is associated with cerebral vasospasm and internal jugular vein may be more adequate vessel for sampling blood to examine these factors. PMID- 9342730 TI - Ether lipid composition and molecular species alterations in carp brain (Cyprinus carpio L.) during normoxic temperature acclimation. AB - Carp (Cyprinus carpio L.) whole brain was used to investigate the thermal acclimation changes under normoxic conditions of three-subclasses (alkenylacyl-, alkylacyl- and diacyl-subclasses) of choline glycerophospholipids (CGP), ethanolamine glycerophospholipids (EGP) and inositol glycerophospholipids (IGP) as well as their acyl chain profiles and molecular species composition. The alkenylacyl subclass of CGP and IGP and the alkylacyl subclass of CGP and EGP varied significantly during summer (25 degrees C) acclimation compared to winter (5 degrees C). The levels of alkenylacyl and alkylacyl-CGP, alkylacyl-EGP and alkenylacyl-IGP were 17.3-, 3.7-, 3.5- and 1.3-fold higher in the summer, respectively, while the alkenylacyl EGP was moderately lower. The levels of diacyl subclasses from CGP and IGP were considerably lower in the summer to compensate for the higher proportion of alkenylacyl and alkylacyl subclasses. Significant changes of ether phospholipids and the reorganization of the molecular species composition of all lipid subclasses may be associated with the "fine tuning" of the physical properties of the cellular membranes in carp brain due to temperature acclimation. The overall acyl chain profile of the three subclasses of carp brain phospholipids showed differences in composition depending upon the subclass of the individual phospholipid. Generally the polyunsaturated fatty acid (PUFA) chain composition increased relative to monounsaturated fatty acid (MUFA) and saturated fatty acids (SFA) during winter acclimation. Docosahexaenoic acid (DHA) was richer in the winter compared to summer. However, no DHA was found in ether-containing species of IGP from either winter or summer, except for 2% in alkylacyl-IGP during the summer. The above observations suggest that the content of ether phospholipids (alkenylacyl and alkylacyl) as well as the reorganization of the molecular species composition of all phospholipids may serve to maintain a functional fluid-crystalline state to preserve the signaling functions in carp brain. PMID- 9342731 TI - Nitric oxide synthase inhibitors do not attenuate diacylglycerol or monoacylglycerol lipase activities in synaptoneurosomes. AB - Neuron-enriched cultures and synaptoneurosomal fractions from 10 day-old rat brain contain diacylglycerol and monoacylglycerol lipase activities. Glutamate and its analogs stimulate the activities of diacylglycerol and monoacylglycerol lipases in a time- and dose-dependent manner. Stimulation of diacylglycerol and monoacylglycerol lipases by glutamate or NMDA can be blocked by MK-801 (non competitive antagonist). Nitro L-arginine methyl ester and L-methylarginine have no effect on glutamate stimulated activities of diacylglycerol and monoacylglycerol lipases. Our studies suggest that synaptoneurosomal preparations from young rat brain are useful for obtaining important information on signal transduction. PMID- 9342732 TI - Alterations in gene expression associated with primary demyelination and remyelination in the peripheral nervous system. AB - Primary demyelination is an important component of a number of human diseases and toxic neuropathies. Animal models of primary demyelination are useful for isolating processes involved in myelin breakdown and remyelination because the complicating events associated with axonal degeneration and regeneration are not present. The tellurium neuropathy model has proven especially useful in this respect. Tellurium specifically blocks synthesis of cholesterol, a major component of PNS myelin. The resulting cholesterol deficit in myelin-producing Schwann cells rapidly leads to sychronous primary demyelination of the sciatic nerve, which is followed by rapid synchronous remyelination when tellurium exposure is discontinued. Known alterations in gene expression for myelin proteins and for other proteins involved in the sequence of events associated with demyelination and subsequent remyelination in the PNS are reviewed, and new data regarding gene expression changes during tellurium neuropathy are presented and discussed. PMID- 9342734 TI - Dietary alpha-linolenic acid increases the biosynthesis of the choline glycerophospholipids from [14C]CDPcholine in rat liver and kidney but not in brain. AB - The effect of feeding rats for 30 days with diets containing high levels of linoleic acid (sunflower oil, SO) or alpha-linolenic acid (perilla oil, PO) was studied in the liver, kidney and brain. The PO group showed a higher labeling of choline glycerophospholipids (CGP) in liver and kidney but no difference with the SO group in ethanolamine glycerophospholipids (EGP) labeling. The brain displayed the lowest incorporation of both precursors and no difference between the two diets. Analyses of brain CGP and EGP fatty acid composition showed that in the PO group the ratio n-6/n-3 was lower than in the SO group, mainly as a consequence of lower levels of n-6 fatty acids. The mole % of docosahexaenoate (DHA) in these lipids was the same for both groups and only triacylglycerols (TAG) displayed a higher DHA. Therefore, at least in the brain, the magnitude of fatty acid changes observed in CGP and EGP for the PO group does not affect the uptake/incorporation of the precursors into phospholipids. PMID- 9342733 TI - Opioid receptor and calcium channel regulation of adenylyl cyclase, modulated by GM1, in NG108-15 cells: competitive interactions. AB - GM1 ganglioside was previously shown to function as a specific regulator of excitatory opioid activity in dorsal root ganglion neurons and F11 hybrid cells, as seen in its facilitation of opioid-induced activation of adenylyl cyclase and its ability to dramatically reduce the threshold opioid concentration required to prolong the action potential duration. The elevated levels of GM1 resulting from chronic opioid exposure of F11 cells were postulated to cause the ensuing opioid excitatory supersensitivity. We now show that GM1 promotes opioid (DADLE)-induced activation of adenylyl cyclase in NG108-15 cells which possess the delta-type of receptor. In keeping with previous studies of other systems, this can be envisioned as conformational interaction of GM1 with the receptor that results in uncoupling of the receptor from Gi and facilitated coupling to Gs. This would also account for the observation that DADLE-induced attenuation of forskolin stimulated adenylyl cyclase was reversed by GM1, provided the cells were not pretreated with pertussis toxin. When the cells were so pretreated, GM1 evoked an unexpected attenuation of forskolin-stimulated adenylyl cyclase attributed to GM1 promoted influx of calcium which was postulated to inhibit a calcium-sensitive form of adenylyl cyclase. This is concordant with several studies showing GM1 to be a potent modulator of calcium flux. Pertussis toxin in these experiments exerted dual effects, one being to promote interaction of the delta-opioid receptor with Gs through inactivation of Gi, and the other to enhance the GM1 promoted influx of calcium by inactivation of Go; the latter is postulated to function as constitutive inhibitor of the relevant calcium channel. NG108-15 cells thus provide an interesting example of competitive interaction between two GM1-regulated systems involving enhancement of both opioid receptor excitatory activity and calcium influx. PMID- 9342737 TI - In vitro phagocytosis of Giardia duodenalis cysts by hemocytes of the Asian freshwater clam Corbicula fluminea. AB - Hemocytes of the Asian freshwater clam Corbicula fluminea, phagocytosed in vitro infectious Giardia duodenalis cysts. After 15, 30, 60, 90, and 120 min of incubation an average of 22%, 32%, 43%, 54%, and 72% of the cysts were phagocytosed by 22%, 55%, 63%, 81%, and 86% of the hemocytes, respectively. The number of hemocytes showing phagocytosis and the mean number of cysts ingested per hemocyte increased significantly over time (P < 0.01); the numbers of nonphagocytosed cysts significantly decreased (P < 0.02). Extrapolation reveals that C. fluminea can retain by phagocytosis an average of 1.6 x 10(6) G. duodenalis cysts/ml hemolymph. The phagocytic capacity of C. fluminea hemocytes indicates the applicability of this freshwater benthic bivalve for bioindication of contamination of waste waters and agricultural drainage with Giardia cysts. PMID- 9342735 TI - Activities of enzymes involved in the metabolism of platelet-activating factor in neural cell cultures during proliferation and differentiation. AB - Platelet-Activating Factor (PAF) is a potent lipid mediator involved in physiological and pathological events in the nervous tissue where it can be synthesized by two distinct pathways. The last reaction of the de novo pathway utilizes CDPcholine and alkylacetylglycerol and is catalyzed by a specific phosphocholinetransferase (PAF-PCT) whereas the remodelling pathway ends with the reaction catalyzed by lyso-PAF acetyltransferase (lyso-PAF AcT) utilizing lyso PAF, a product of phospholipase A2 activity, and acetyl-CoA. The levels of PAF in the nervous tissue are also regulated by PAF acetylhydrolase that inactivates this mediator. We have studied the activities of these enzymes during cell proliferation and differentiation in two experimental models: 1) neuronal and glial primary cell cultures from chick embryo and 2) LA-N-1 neuroblastoma cells induced to differentiate by retinoic acid (RA). In undifferentiated neuronal cells from 8-days chick embryos the activity of PAF-PCT was much higher than that of lyso-PAF AcT but it decreased during the period of cellular proliferation up to the arrest of mitosis (day 1-3). During this period no significant changes of lyso-PAF AcT activity was observed. Both enzyme activities increased during the period of neuronal maturation and the formation of cellular contacts and synaptic like junctions. The activity of PAF acetylhydrolase was unchanged during the development of the neuronal cultures. PAF-PCT activity did not change during the development of chick embryo glial cultures but lyso-PAF AcT activity increased up to the 12th day. RA treatment of LA-N-1 cell culture in proliferation decreased PAF-PCT activity and had no significant effect on lyso-PAF AcT and PAF acetylhydrolase indicating that the synthesis of PAF by the enzyme catalyzing the last step of the de novo pathway is inhibited when the LA-N-1 cells are induced to differentiate. These data suggest that: 1) in chick embryo primary cultures, both pathways are potentially able to contribute to PAF synthesis during development of neuronal cells particularly when they form synaptic-like junctions whereas, during development of glial cells, only the remodelling pathway might be particularly active on synthesizing PAF; 2) in LA-N-1 neuroblastoma cells PAF synthesizing enzymes coexist and, when cells start to differentiate the contribution of the de novo pathway to PAF biosynthesis might be reduced. PMID- 9342736 TI - Effects of IL-1 beta on receptor-mediated poly-phosphoinositide signaling pathway in immortalized astrocytes (DITNC). AB - Astrocytes are known to play multi-functional roles in support of many homeostatic mechanisms in the central nervous system including host defense mechanisms. Despite the ability of cytokines to alter gene expression and cellular activity, their effect on receptor-mediated poly-phosphoinositide (poly PI) signaling pathway has not been examined in detail. In this study, an immortalized astrocyte cell line (DITNC) was used to test the effect of IL-1 beta exposure on the poly-PI signaling pathway. Similar to primary astrocytes, DITNC cells exhibit P2-purinergic receptor response to ATP and UTP leading to transient increases in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and intracellular calcium concentration, [Ca2+]i. Upon exposure of DITNC cells to IL-1 beta (100 U/ml) for 24 hrs, an increased response to the poly-PI agonists was observed. The increase in ATP-mediated Ins(1,4,5)P3 release could not be attributed to a shift in the ATP dose or an alteration of the time profile for the release of Ins(1,4,5)P3. Since the increase in response required a lag time of 4 hr after IL 1 beta exposure, it is unlikely that this effect was due to a direct interaction of IL-1 beta with the purinergic receptor. On the other hand, an increase in ATP response could be observed in DITNC cells exposed to conditioned medium obtained after IL-1 beta treatment. It can be concluded that exposure of astrocytes to cytokines may lead to an increase in receptor-mediated poly-PI signaling activity and this may involve compounds secreted into the culture medium, e.g., the secretory phospholipase A2. PMID- 9342738 TI - Recognition of the parasite infected cell surface determinants by homologous antiserum raised against infected cell membranes. AB - Identification of neo-antigenic determinant(s) on parasite infected cell surface is important to control intracellular infections. Such determinant(s) on the surface of intact Plasmodium berghei infected erythrocytes have not been conclusively demonstrated. To generate polyclonal antiserum selectively recognizing the parasite infected cell surface determinant(s), in natural state, we have examined the efficacy of the homologous immunizations, in BALB/c mice, with the membrane rich preparation of: i) erythrocytes in vivo infected with Plasmodium berghei and, ii) macrophages in vitro infected with Leishmania donovani. Anti-infected erythrocyte membrane antiserum specifically recognized, albeit at low level, the infected cell surface as determined by flow cytometry and immunoelectron microscopy. Immunoprecipitation of radiolabeled antigens revealed at least three parasite proteins of > 205 kDa, 160 kDa and 100 kDa specifically present on infected erythrocyte surface. Normal uninfected erythrocytes did not react with the antiserum. Anti-L. donovani-infected macrophage membrane antiserum also recognized only infected macrophage surface and not the normal macrophages. Thus, the approach may find wide application in delineating disease specific determinant(s) on the infected cell surface, particularly to those where animal models are available. PMID- 9342739 TI - Monoclonal antibodies against diagnostic Anisakis simplex antigens. AB - Five monoclonal antibodies (UA2, UA3, UA5, UA6, and UA8) specific for Anisakis simplex are described. All are IgG1/kappa monoclonal antibodies, except for UA2, which is an antibody IgM/kappa. The molecular weights of the major components recognized in immunoblotting are 48 and 67 kDa (UA2); 139 kDa (UA3 and UA5; same epitope); 35, 38, and 139 kDa (UA6); and 205 kDa (UA8). UA2 was the only monoclonal antibody to recognize both components of an excretion-secretion antigen preparation and antigens in the excretory cell and esophageal glands of third-stage A. simplex larvae; antigens in the excretory cell were also recognized by UA3 and UA6. Cross-reactivity studies using a hyperimmune polyclonal rabbit serum reacting with various ascaridoid nematodes indicated that the antigens captured by our monoclonal antibodies were specific for A. simplex. Finally, comparative studies of our monoclonal antibodies and An2 (the only monoclonal antibody currently available for serodiagnosis of human anisakiasis), based on the calculation of multiples of normal activity for human anisakiasis sera, indicated that our monoclonal antibodies (and particularly UA3) recognized antigens that are good candidates for serodiagnostic purposes. PMID- 9342740 TI - Nitric oxide involvement in experimental Trypanosoma cruzi infection in Calomys callosus and Swiss mice. AB - Nitric oxide (NO) production by peritoneal macrophages was evaluated in Calomys callosus and Swiss mice during the course of infection with two strains of Trypanosoma cruzi. In C. callosus, no NO production was detected throughout the period of observation in animals infected with either parasite strain, except for a very low amount measured on day 40 in animals infected with strain M226 and on the 28th day in animals infected with strain F after in vitro stimulation with interferon gamma (IFN-gamma). Macrophages of Swiss mice produced large amounts of NO, the highest values being observed on the 40th day in mice infected with the F strain. Induced nitrogen oxide synthase (iNOS) was not detected in macrophages of infected C. callosus but was detected in mice. The i.p. inoculation of thioglycolate, bacille Calmette-Guerin (BCG) and periodate, nonspecific macrophage activators, did not induce NO production in C. callosus, but high levels were observed in Swiss mice after secondary in vitro IFN-gamma plus lipopolysaccharide (LPS) stimulation. However, H2O2 release was induced in macrophages stimulated with phorbol myristate acetate (PMA) in both experimental models. Serum NOx(NO2 + NO3) levels were low in C. callosus infected with strain M226, which was originally isolated from this animal species. Strain-F-infected animals had higher serum NOx levels in the initial period of infection, which dropped to noninfected control values on the 40th day. In Swiss mice, both strains induced the production of higher levels of NOx throughout the period of observation, with the increase being more pronounced in mice infected with the F strain. Daily treatment of F-strain-infected C. callosus with the arginine analogue L-nitro-arginine drastically reduced NOx levels, with no influence on parasitemia or mortality being observed. The results obtained suggest that C. callosus shows a distinct behavior with regard to resistance to T. cruzi infection. PMID- 9342741 TI - Induction of stress proteins in the plant trypanosome Phytomonas characias. AB - The present study of the synthesis of new proteins in plant trypanosomatids in the genus Phytomonas as a response to different types of stress demonstrates the production of a number of proteins that can be grouped into four families similar to those that appear in other organisms (heat-shock proteins). In the study of stress, Phytomonas cultures were subjected to changes in temperature from 22 degrees to 37 degrees C, deprived of glucose, grown in the presence of sodium arsenite, and treated with calcium ionophore. In addition, the culture medium was changed from Grace's medium (330 mosmol/1) to a plant-culture medium with an osmolarity of 286 mosmol/l, implying the exertion of stress during the parasite's normal biological cycle of passage from the insect vector to the plant host. The treatment with actinomycin D demonstrated that some of the mRNAs that codify these proteins are found in normal presynthesized conditions. To measure the effect of temperature on the macromolecule biosynthesis we compared the incorporation of labeled analogues ([3H]-thymidine, [3H]-uridine, and [3H] leucine) by flagellates cultured at 22 degrees C with that by parasites cultivated at 37 degrees C. PMID- 9342742 TI - Babesia bovis: identification of immunodominant merozoite surface proteins in soluble culture-derived exoantigen. AB - Babesia bovis merozoite proteins presenting as exoantigens in in vitro culture supernatants have been characterized. Bovine antisera to B. bovis exoantigens were used to immunoprecipitate [35S]-methionine metabolically labeled or lactoperoxidase-catalyzed radioiodinated B. bovis merozoite proteins. A total of 24 metabolically labeled proteins ranging in molecular weight from 24,000 to 225,000 Da and 9 radioiodinated proteins with molecular weights varying between 24,000 and 225,000 Da were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Monoclonal antibodies to B. bovis merozoite surface proteins were also used to immunoprecipitate metabolically labeled exoantigens directly from in vitro culture supernatants. These results demonstrate epitopes from at least nine merozoite surface proteins present in the exoantigen fraction, among which are the recently characterized major surface antigens 1 and 2, rhoptry associated protein 1, and spherical body protein 2. PMID- 9342743 TI - Trypanosoma cruzi in the rectum of the bug Triatoma infestans: effects of blood ingestion of the vector and artificial diuresis. AB - The population density and the percentage of different developmental stages of an established infection of Trypanosoma cruzi were determined at 40 days after the last feeding of the fourth instar in the rectum (lumen, anterior and posterior wall) of fed and unfed groups of fifth instars of Triatoma infestans. Additionally, the rectum and the Malpighian tubules were incubated in saline, inducing diuresis by addition of the diuretic hormone. The rectum contained an average of 200,000-400,000 T. cruzi. After feeding the percentages of spheromastigotes and drop-like intermediate stages were reduced from < 7% and 15%, respectively, to < 3%, but those of slender intermediate stages increased statistically significantly from < 7% to 10%. After 4 h of diuresis the in-vitro incubated isolated rectum with the four Malpighian tubules showed the same trends, indicating that diuresis rather than factors of the hemolymph or digestive products induces the development of metacyclic trypomastigotes of T. cruzi originating from epimastigotes. PMID- 9342744 TI - Seasonal differences in infestation of the perch Perca fluviatilis L. from Lake Constance with digenean trematodes. AB - Perch (Perca fluviatilis L.) caught in Lake Constance every 2 months over a period of 3 years below six towns or villages (Langenargen, Nonnenhorn, Rorschach, Romanshorn, Bottighofen, and the Lake of Uberlingen) were examined for parasites. In contrast to Diplostomum spathaceum (Rudolphi, 1819) and Tylodelphys clavata (Nordmann, 1832), Bunodera luciopercae (Muller, 1776) and Ichthyocotylurus variegatus (Creplin, 1825) showed marked seasonal differences. These differences were influenced by various factors: the different numbers of the first intermediate hosts (snails, copepods), the water temperature, the physiological state of the fish, and its way of life. PMID- 9342745 TI - The parasitism of fish from Lake Constance--a comparison of present and earlier data. AB - Commercial fish (perch, roach, bream, dace, and burbot) were caught below Langenargen in 1988-1990. Their infestation with cestodes and digenean trematodes was studied. The results were compared with earlier data. The infestation of perch with Diplostomum spathaceum and Ichthyocotylurus variegatus noted in 1988 1990 had increased as compared with that observed in 1974-1976. The infestation of perch with Triaenophorus nodulosus seen in 1990 had increased as opposed to that noted in 1988. The infestation of bream with D. spathaceum and of perch and roach with Tylodelphys clavata observed in 1988-1990 had remained the same as that seen in 1974-1976. The infestation of perch with D. spathaceum noted in 1988 1990 had decreased as opposed to that reported for 1974-1976. The infestation of bream, dace, and burbot with T. clavata noted in 1988-1990 had decreased as compared with that observed in 1974-1976. The infestation of perch with T. nodulosus seen in 1988/1989 had decreased as opposed to that noted in 1974-1976. All three cyprinids studied as well as the burbot had been less intensively infested in 1988-1990 than in 1974-1976. Only perch had been more intensively infested in 1988-1990 than in 1974-1976, especially with D. spathaceum and I. variegatus. PMID- 9342746 TI - Trypanosoma rangeli: increase in virulence with inocula of different origins in the experimental infection in mice. AB - We compared two murine models of Trypanosoma rangeli infection. The same inoculum dose and age-matched hosts were used in both cases. One group was infected with trypomastigotes obtained from passages in mice and the other, with trypomastigotes obtained from cell culture after a passage in mice. We observed that trypomastigotes obtained from the in vitro cellular infection showed increased virulence in experimental animals, with a 70% rate of death being noted in experimental mice instead of the lack of mortality seen when in vivo-derived parasites were used. The greatest levels of parasitemia and tissual lesions in the presence of the parasite also occurred when in vitro-derived parasites were used. PMID- 9342747 TI - Cell-surface carbohydrates of Entamoeba invadens. AB - Cell-surface carbohydrates of Entamoeba invadens trophozoites were analyzed using (a) a panel of highly purified lectins specific for molecules containing N acetylglucosamine or sialic acid, N-acetylgalactosamine, galactose, mannose-like residues, and fucose; (b) Escherichia coli K-12 with mannose-sensitive fimbria; (c) enzymatic digestion; and (d) scanning electron microscopy. The presence of galactose (D-Gal) and N-acetylgalactosamine (D-GalNAc) was detected in the amoeba. Previous trypsinization induced the appearance of Glycine max (SBA, specific for D-GalNAc residues)-binding sites, whereas such treatment completely abolished the ability of Ricinus communis (RCAI) and Axinalla polypoides (APP, specific for D-Gal) lectins and partially abolished that of Euonymus europaeus (EEL, specific for D-Gal) lectins to agglutinate the trophozoites. The agglutinating activity of E. coli K-12 adheans with the amoeba was markedly increased after trypsin digestion, indicating that mannose units become exposed after enzyme treatment. These findings were essentially confirmed by scanning electron microscopy. After neuraminidase treatment the parasites became strongly agglutinated with SBA and Arachis hypogaea (PNA, specific for D-Gal) and the cell interaction with Wisteria floribunda (WFH, specific for D-GalNAc) was markedly increased. These results suggest that in E. invadens trophozoites, sialic acid residues are linked to D-Gal and D-GalNAc. PMID- 9342748 TI - Miracidial host-finding in Fasciola hepatica and Trichobilharzia ocellata is stimulated by species-specific glycoconjugates released from the host snails. AB - The miracidia of Fasciola hepatica and Trichobilharzia ocellata approach their host snails Lymnaea truncatula and L. stagnalis by increasing their rate of change of direction (RCD) in increasing gradients of snail-conditioned water (SCW), and they perform a turnback swimming in decreasing gradients. Both hostfinding responses in both species were induced by glycoconjugates with a molecular weight of > 30 kDa that were sensitive to hydrolysis with pronase E and oxidation with NaIO4. Alkaline cleavage revealed that they contained carbohydrates linked O-glycosidically via serine and N-acetylgalactosamine. Miracidia clearly preferred SCW from their specific host snail versus other sympatric snail species and did not respond to water conditioned with fish, tadpoles, or leeches. Differences in the chemical characteristics of SCW from the intermediate hosts L. truncatula and L. stagnalis could be shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotting, and subsequent carbohydrate detection. The first step of purification of the effective signaling SCW components from both snail species was achieved by ion-exchange chromatography. PMID- 9342749 TI - The use of inhibitors of N-linked glycosylation and oligosaccharide processing to produce monoclonal antibodies against non-phosphorylcholine epitopes of Brugia pahangi excretory-secretory products. AB - Adult Brugia pahangi were cultured with [3H]-choline in both the absence and the presence of either tunicamycin or 1-deoxymannojirimycin (dMM), inhibitors of N linked glycosylation and N-linked oligosaccharide processing, respectively. Excretory-secretory products (ES) were recovered from the spent medium and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/fluorography. Both inhibitors were found to prevent radiolabeling of ES, a result consistent with blockage of the addition of the highly immunodominant phosphorylcholine (PC) group. Tunicamycin/dMM-treated ES were subsequently employed to immunise a mouse in an attempt to produce monoclonal antibodies (mAbs) against non-PC epitopes of ES. Three mAbs were isolated, each of which reacted with ES but not with PC. PMID- 9342750 TI - Susceptibility of dyskinetoplastic Trypanosoma evansi and T. equiperdum to isometamidium chloride. AB - Isometamidium chloride (ISM) is an effective trypanocide with curative and prophylactive activity in husbandry animals. The mode of action of ISM against pathogenic trypanosomes is not fully understood, but there is evidence in the literature that kinetoplastic topoisomerase type II is selectively inhibited by the drug. This prompted the hypothesis that dyskinetoplastic trypanosomes would express a reduced level of susceptibility to ISM. From parental Trypanosoma evansi and T. equiperdum populations we generated clones containing only dyskinetoplastic organisms and clones containing organisms with kinetoplasts. The susceptibility of these clones to ISM was quantified by in vitro assays. The susceptibility of all clones was in the same range. Minor differences in drug susceptibility found between the clones showed that the dyskinetoplastic T. evansi and T. equiperdum clones were even more susceptible to ISM than their kinetoplastic counterparts. Thus, the hypothesis that the dyskinetoplastic trypanosomes would be less susceptible to or tolerant of ISM was clearly disproved. The previously demonstrated inhibition of kinetoplastic topoisomerase type II by ISM cannot be the primary toxic effect of the drug on trypanosomes, and the mode of action of ISM needs to be reassessed. PMID- 9342751 TI - Morphometric determination of the methodological criteria for the diagnosis of intestinal neuronal dysplasia (IND B). AB - Intestinal neuronal dysplasia of the submucous plexus (IND B) is an indicator of a developmental abnormality of vegetative gut innervation. It is the mildest form of an inborn error of intestinal innervation. The diagnosis of IND B does not result in a functional conclusion or clinical recommendation but is often accompanied by oligoneuronal hypoganglionosis of the myenteric plexus or an aganglionosis of the rectum. The aim of this study was to demonstrate by morphometric means a way in which the diagnosis of IND B could be made much more reliable. In 20 control subjects, 40 IND B cases and 10 hypoganglionoses with IND B, it was shown that a specific nerve cell staining (e.g. Lactic dehydrogenase, Succinic dehydrogenase, Diaphorase reaction or an immunohistochemical nerve cell staining) was necessary for diagnosis. Cross sections of giant ganglions and cross sections with large nerve cell numbers (> 7 nerve cell profiles) were the most reliable diagnostic criteria. The morphometric examinations were performed with an optic electronic image analysis system. Biopsy serial sections of the rectum-mucosa that contained submucosa demonstrated that 30-40% of the sections contained no submucous ganglion. Sixty to 70% of the sections showed ganglia of the submucous plexus. In 100 biopsy sections in subjects with IND B, 20 +/- 5% contained giant ganglions cross sections. In the patients with hypoganglionosis of the submucous plexus, 55 +/- 4% sections had no ganglion and 18 +/- 3% had giant ganglion cross sections. The data demonstrate that for a reliable diagnosis of IND B, at least 30 sections are necessary, stained with a dehydrogenase reaction that contain a minimum of 4 giant ganglion cross sections. These data demonstrate that IND B is not a qualitative diagnosis as Hirschsprung's disease but rather a quantitative diagnosis. PMID- 9342752 TI - A comparison of different modes for the detection of p53 protein accumulation. A study of bladder cancer. AB - The aim of the present study was to evaluate different techniques for the analysis of p53 protein accumulation in human bladder cancer. The accumulation was evaluated in 23 carcinomas by immunoblotting (IB), immunohistochemistry (IHC) and flow cytometry (FCM). The results revealed that six (26%), eight (35%) and ten (43%) of the tumours were p53 protein positive by IB, IHC and FCM, respectively. Mutation analysis of the TP53 gene confirmed mutations in 8 of 9 tumours which showed increased levels of p53 protein by FCM. Our results indicate that IHC could be applied for studies of p53 protein accumulation in archival formalin fixed, paraffin-embedded bladder tumours. However, FCM is a more sensitive and objective method for the detection of p53 protein than IHC and this should be taken into account when routinely evaluating the p53 protein accumulation by IHC. PMID- 9342753 TI - Tenascin expression in elastotic cuffs of invasive ductal carcinoma of the breast. AB - We studied immunohistochemically one thousand one hundred and thirty-seven cases of primary invasive breast cancers (NST) and adjacent normal mammary glands for tenascin expression, and compared their elastic content to verify if a relationship exists between tenascin expression and elastosis. Periductal, perivascular and stromal elastosis were graded on a scale from 0 to 3 (absent to massive). All carcinomas showed tenascin expression and elastosis with various histological appearances. In the adjacent breast, teanscon was distributed around the normal ducts or with extasia and uctal hyperplasia without atypia. Digestion of the sections with elastase prior to staining resulted in a loss of the specific staining reactions in all areas where elastosis was present. Tenascin staining was observed in the mesenchyme closely surrounding the neoplastic ducts and the cancer cell nests. Stromal tenascin staining appeared stronger in those carcinomas that exhibited marked desmoplastic reactions. The highly differentiated tumours contained more elastosis in their tumour tissue than the poorly differentiated ones, whereas tenascin expression was stronger in poorly differentiated tumours than well differentiated tumours. A strong staining for tenascin was observed in the elastotic cuff. Tenascin staining did not disappear afterwards with elastase. We did not find a statistically significant correlation between tenascin expression, elastosis and prognostic factors such as size of the tumour, lymph node metastasis, tumour necrosis and age. In our study tenascin proved to be an additional element in elastotic areas even though the significance of an association between elastosis and tenascin is still unknown, as is that of elastosis itself. PMID- 9342754 TI - Linell classification of breast cancer morphology compared to histologic grading, S-phase fraction and DNA-ploidy. AB - One hundred and fifty-eight histologically verified mammary carcinomas with known mammographic doubling time (DT) were studied with special emphasis on a morphologic classification proposed by Linell et al. [8, 12, 14, 15]. The hypothesis that Linell classification of ductal carcinomas into comedo, tubuloductal and tubular carcinomas is easy to perform with small inter-observer variations, was not fully confirmed. The Linell classification was found to correlate well with conventional WHO malignancy grading, S-phase fraction and DNA ploidy. The Linell classification also correlated to surgical stage, lymph node status and DT, but not at all to tumour size. Using distant disease-free survival as an endpoint, the Linell classification gave prognostic information comparable to conventional histologic grading, seeming to be a simple, cheap and reliable method well worth trying on a larger scale. PMID- 9342755 TI - Inflammatory response in cervical intraepithelial neoplasia and squamous cell carcinoma of the uterine cervix. AB - Leukocytic infiltrates are a morphologic feature of most solid tumors, including uterine cervical intraepithelial neoplasia (CIN) and invasive carcinoma. We have studied 50 cases of CIN I, CIN II, CIN III, invasive carcinoma and normal controls in order to evaluate the inflammatory response. Two markers--CD68, a macrophage-specific marker, and ICAM-1, present on leukocytes, blood vessels and epithelial cells--were employed. Results have demonstrated similar inflammatory cell counts in normal, CIN II and CIN III lesions by both markers, and lower counts for CIN I. Invasive carcinomas demonstrated a statistically significant increase in infiltrate density by both CD68 (p < 0.002) and ICAM-1 (p < 0.05). Macrophage density by either marker did not correlate with Human Papillomavirus (HPV) presence, specific type, or evidence of co-infection with several types. We conclude that the inflammatory response to cervical intraepithelial-neoplasia is inadequate. The elevated cell counts in invasive carcinomas may reflect a reaction towards invasion rather than tumor-specific immune response. Depression of inflammation in CIN I lesions may be associated with active viral replication in these lesions. PMID- 9342756 TI - Comparative immunoreactivity of CD-68 and HMB-45 in malignant melanoma, neural tumors and nevi. AB - The monoclonal antibody CD 68 (KP 1) reacts with fibrohistiocytic and some epithelial neoplasms; its reactivity compared with that of HMB 45 in malignant melanoma (MM) and neural tumors needs further elucidation. Using a streptavidin biotin-immunoperoxidase procedure, we examined the reactivity of 65 MM (46 conventional, 1 polypoid, 6 desmoplastic [DMM], and 12 metastatic), 21 neurofibromas, 1 neurofibrosarcoma, 10 schwannomas, 1 perineurioma, 2 neurothekeomas, and 14 blue and 26 other nevi for CD-68, HMB-45-defined antigen, S 100 and neurofilament protein. A positive staining for CD 68 was observed in 38 of 42 primary, 5 of 6 DMM, and 11 of 12 metastatic melanomas; 6 of 10 schwannomas; 5 of 10 nevi with junctional component and all 14 blue nevi. All 21 neurofibromas, 1 each neurofibrosarcoma and perineurioma, both neurothekeomas, and all 12 nevi with dermal component were CD 68-negative. HBM 45 was expressed by all 44 primary, none of 6 DMM, and 7 of 12 metastatic melanomas; by none of 10 schwannomas, 6 neurofibromas, 1 neurofibrosarcoma, 1 perineurioma and 2 neurothekeomas. Both junctional nevi, 8 of 10 nevi with junctional components, 1 of 10 dermal components of junctional nevi, and 11 of 13 blue nevi were also HMB 45 positive. Except for 1 perineurioma, S 100 decorated all tumors examined. NF was immunoreactive in 1 of 45 conventional melanomas, 2 of 21 neurofibromas, 2 of 10 schwannomas, and 3 of 10 blue nevi; it was non-reactive in all polypoid, desmoplastic and metastatic melanomas; neurofibrosarcoma, perineurioma, neurothekeoma and other nevi. We conclude that the CD-68-reactivity in primary melanomas, neurofibromas, neurofibrosarcomas, perineuriomas, and nevi was similar to that of HMB 45. The significantly higher CD 68-positivity than of HMB 45 in metastatic and desmoplastic melanomas and schwannomas may be of diagnostic value. PMID- 9342757 TI - Immunohistochemical detection of vascular growth factors in angiomatous and atypical meningiomas, as well as hemangiopericytomas. AB - The arachnoideal compartment provides the vascular sources for three different tumor types rich in vessels: angiomatous meningioma, some atypical meningioma with high vascularity and meningeal hemangiopericytoma. We investigated immunohistochemically the expression and distribution of vascular mitogenes in 7 angiomatous meningiomas, 8 atypical meningiomas with high vasculature and 4 hemangiopericytomas. On the one hand it should be studied which vascular growth factors such as VPF/VEGF-1, VPF/VEGF-2, bFGF, PDGF and TGF-alpha could be responsible for the close meshwork of vessels within the tumors. On the other hand we were interested in whether or not there are differences in vascular mitogens between slowly growing angiomatous meningiomas and both other types with their increased tendency to recur. PDGF and TGF-alpha were extensively expressed in the endothelium and smooth muscle cells of the vessels, as well as tumor cells. VEGF-2 could only be found in endothelial cells of all three tumor entities. bFGF was localized in some vessels of angiomatous meningiomas and VEGF 1 revealed a very low expression with a localization comparable with VEGF-2. Moreover, uPAR was diffusely expressed in nearly all tumor cells and endothelial cells. The fact that tumor cells of hemangiopericytomas and meningiomas did not show any immunohistochemical reaction with VEGF's could indicate a lower priority of these growth factors for neovascularization in this type of neoplasm. A different expression of vascular mitogens between benign angiomatous meningiomas and atypical meningiomas as well as hemangiopericytomas with their tendency for recurrence could not be observed. The morphological evidence for extravasates of IgG-proteins, Fibrin and Fibronectin due to VPF-effects seems not to be a renouncable condition for neoangiogenesis in the tumors investigated. PMID- 9342758 TI - In vitro cytokine secretion and maturation phenotype of lymphoma-associated splenic macrophages. AB - In this study, cytokine secretion capacity and maturation phenotype of human lymphoma-associated splenic macrophages (LASM) were evaluated in a long-term culture. Sixteen spleens from malignant lymphoma patients and five control spleens were investigated. Splenic macrophages (SM) were isolated by teflon adherence and cultured for 6-48 days. Secretion of IL-1 alpha, IL-6 and TNF alpha was measured by ELISA following maximal stimulation with LPS and IFN-gamma, and cytokine mRNA expression was detected by in situ hybridization. Immunohistochemical expression of maturation-associated antigens was evaluated semiquantitatively. Cytokine secretion capacity was significantly altered in LASM which exhibited reduced TNF alpha and IL-6, but elevated IL-1 alpha secretion when compared to control SM. Alterations of cytokine secretion capacity were associated with a modification of LASM maturation phenotype, showing impaired expression of early and chronic/late inflammatory markers. These findings obtained from a long-term culture model suggest that malignant lymphomas induce lasting modifications of cytokine secretion and maturation patterns in LASM. PMID- 9342759 TI - Hepatic hilar inflammatory pseudotumor mimicking cholangiocarcinoma with cholangitis and phlebitis--a variant of primary sclerosing cholangitis? AB - Inflammatory pseudotumor (IPT) of the liver is rare. We present a case of hepatic IPT mimicking cholangiocarcinoma in which the tumor was located at the left porta hepatis. The patient was a 64-year-old man in whom abnormal liver function test results had been noted incidentally during an annual health checkup in 1993: the patient declined to go to the hospital for further examination. At the annual health checkup the following year, abnormal liver function test results were noticed again, and this time he did go to a hospital, where a hepatic mass was found. Laboratory test results were unremarkable. Based on the location of the lesion and the findings of a variety of imaging modalities, such as ultrasound and computed tomography examination, the lesion was preoperatively diagnosed as hilar cholangiocarcinoma and was surgically resected. Pathologic examination of the resected lesion, however, revealed that it was not a true tumor but an inflammatory pseudotumor with marked destructive and sclerosing cholangitis mimicking primary sclerosing cholangitis (PSC) and obliterative phlebitis. Since the location and features of the tumor in the present case are very pertinent to the relationship between IPT and PSC, we describe its clinical and histologic features and discuss the findings in relation to PSC in the context of our literature review. PMID- 9342760 TI - Case report: massive cardiopulmonary cysticercosis in a leukemic patient. AB - A 53-year-old woman who had acute leukemia and massive cardiopulmonary cysticercosis is reported. Pulmonary cysticercosis is rare and in this case may have been promoted by the immunosuppression caused by acute leukemia. PMID- 9342761 TI - Multilocular thymic cysts associated with thymoma: a case report. PMID- 9342762 TI - Clinically used antidepressant drugs. PMID- 9342763 TI - Controlled clinical trials of hypericum extracts in depressed patients--an overview. AB - In Germany, hypericum extracts are among the most widely prescribed antidepressants. Additionally, many preparations of St. John's wort are sold on the free market and one extract is even the best selling antidepressant in the country. In contrast to synthetic antidepressants, the approval procedures are not so strict, which implies that the pharmaceutical industry is not forced to conduct clinical trials suitable for licensing. Nevertheless, numerous studies on hypericum extracts including depressed patients have been published in the last 20 years. The purpose of this paper is to review these investigations in respect of methodological considerations and to draw conclusions pertaining to the proof of antidepressant efficacy. To this effect, a computer-assisted literature research was performed and manufacturers were asked to supply the author with study results. A total of 12 placebo-controlled trials with hypericum extracts were performed, mostly with positive results. Also in comparison with synthetic antidepressants (3 studies published), a similar reduction of depressive symptomatology was seen, although the comparators were not adequately dosed. No trials in severely depressed patients have been published yet. Since most studies on hypericum have methodological flaws, further studies are warranted. PMID- 9342764 TI - LI 160, an extract of St. John's wort, versus amitriptyline in mildly to moderately depressed outpatients--a controlled 6-week clinical trial. AB - Up to now, the antidepressant efficacy of the extract of St. John's wort, LI 160, has been compared to imipramine and maprotiline, demonstrating similar antidepressant efficacy in mildly to moderately depressed patients, treated either with LI 160 or the respective synthetic comparator. In the study reported here, LI 160 (total daily dose: 900 mg) was compared with the sedating tricyclic amitriptyline (total daily dose: 75 mg) in a controlled, randomized, multicentre trial. At the end of the 6-week study, the major target variable, the Hamilton Depression Scale response rate, exhibited no statistically significant difference between the groups, although a tendency for a better response rate was seen in the amitriptyline group. The secondary efficacy parameters, decreases in the total Hamilton Depression and Montgomery-Asberg scores, showed a significant advantage for amitriptyline, but only at week 6. With regard to tolerability, LI 160 was clearly superior to amitriptyline, particularly in relation to anticholinergic and Central Nervous System adverse events. Thus, 37% of the LI 160 treated patients reported adverse events, compared to 64% in the amitriptyline group. This considerable superiority in tolerability for LI 160 in relation to amitriptyline, could confer an advantage in improving compliance for antidepressant pharmacotherapy. PMID- 9342765 TI - Efficacy and tolerability of St. John's wort extract LI 160 versus imipramine in patients with severe depressive episodes according to ICD-10. AB - The special extract of St. John's wort, LI 160, exhibited a superior antidepressant efficacy compared to placebo in several controlled trials. Two further trials demonstrated a similar reduction of depressive symptomatology under LI 160 compared to tricyclics. All these trials were performed in mildly to moderately depressed patients. The present investigation was a randomized, controlled, multicentre, 6-week trial comparing 1800 mg LI 160/die to 150 mg imipramine/die in severely depressed patients according to ICD-10. The main efficacy parameter, a reduction of the total score of the Hamilton Depression Scale, proved both treatment regimens very effective at the end of the 6 week treatment period (mean values 25.3 to 14.5 in the LI 160 group and 26.1 to 13.6 in the imipramine group), but not statistically equivalent within a a-priori defined 25% interval of deviation. The analysis of subgroups with more than a 33% and 50% reduction of the HAMD total score justified the assumption of equivalence within a 25% deviation interval. This view was also supported by the global efficacy ratings from patients and investigators. Regarding adverse events, the nonrejection of the nonequivalence hypothesis denotes a superiority of the herbal antidepressant. These main result indicate that LI 160 might be a treatment alternative to the synthetic tricyclic antidepressant imipramine in the majority of severe forms of depressions. However, more studies of this type must be performed before a stronger recommendation can be made. PMID- 9342766 TI - The effect of hypericum extract on cardiac conduction as seen in the electrocardiogram compared to that of imipramine. AB - The electrocardiographic effects of high-dose hypericum extract were compared to the effects of imipramine hydrochloride on ECG recordings in a randomized, double blind, multicenter treatment study of 209 patients suffering from depression. ECGs were recorded before and after a six-week treatment period with either hypericum extract or imipramine. At the end of the study ECGs of 84 patients treated with hypericum extract and 76 patients treated with imipramine were suitable for an analysis of conduction intervals and pathological findings. In the first ECG analysis comparing high dose hypericum extract with imipramine, a prolongation of the conduction intervals PR, QRS and QTc was found for imipramine. In contrast, a small acceleration of conduction was seen for the high dose hypericum extract. The comparison of ECGs at the beginning and after six weeks of treatment showed a significant increase in first degree AV-blocks and abnormalities of repolarization under imipramine but a significant reduction of such pathological findings under treatment with hypericum extract. It should be emphasized that this favorable feature of safe cardiac activity was achieved with 1800 mg of hypericum extract. The reduction in pathological ECG features after treatment with hypericum extract may have resulted mainly from the change of medication, probably tricyclics, to hypericum extract. Our results indicate that for the treatment of patients with a pre-existing conductive dysfunction or elderly patients, high-dose hypericum extract is safer with regard to cardiac function than tricyclic antidepressants. PMID- 9342767 TI - Treatment of seasonal affective disorder (SAD) with hypericum extract. AB - Seasonal affective disorder (SAD) is a subgroup of major depression and characterized by a regular occurrence of symptoms in autumn/winter and full remission or hypomania in spring/summer. Light therapy (LT) and recently pharmacotherapy with specific antidepressants have been shown to be beneficial. Within the array of pharmacotherapy hypericum extract has also been found to be effective in a single-blind study (Martinez et al., 1994). In this 4 weeks treatment study 900 mg of hypericum was associated with a significant reduction in the total score of the Hamilton Depression Rating Scale. There was no significant difference when bright light therapy was combined with hypericum, compared to the situation without bright light therapy. Overall, hypericum was well tolerated and therefore the data suggest that pharmacological treatment with hypericum may be an efficient therapy in patients with SAD, which needs to be substantiated in further controlled studies. PMID- 9342768 TI - Hypericin and pseudohypericin: pharmacokinetics and effects on photosensitivity in humans. AB - Extracts of St. John's wort (Hypericum perforatum) are used in treatment of depression. They contain various substances with the naphthodianthrones hypericin and pseudohypericin as characteristic ingredients. These compounds were shown to cause phototoxicity in cell culture and in animals. A placebo-controlled randomized clinical trial with monitoring of hypericin and pseudohypericin plasma concentration was performed to evaluate the increase in dermal photosensitivity in humans after application of high dose hypericum extracts. The study was divided into a single dose and a multiple dose part. In the single dose period, each of 13 volunteers received in a double blind fourfold complete crossover design, either placebo, or 900, 1800 or 3600 mg of a standardized hypericum extract (LI 160) containing zero, 2.81, 5.62 and 11.25 mg of total hypericin (total hypericin is the sum of hypericin and pseudohypericin). Maximum total hypericin plasma concentrations were observed about 4 h after dosage and were 0, 0.028, 0.061 and 0.159 mg/L, respectively. Before and 4 h after drug intake, the subjects were exposed at small areas of their back to increasing doses of solar simulated irradiation (SSI, with combined ultraviolet A, UV-A, and UV-B light) and another part was exposed to selective UV-A light irradiation. Minimal erythema dose was determined 5, 20 and 68 h after irradiation. Comparison of SSI sensitivity without and with hypericum extract did not show and difference and there was no dose-related trend in light sensitivity. Sensitivity to selective UV A light was increased only after the highest dose from a minimal tanning dose of 10.8 J/ cm2 (mean) after placebo to 8.7 J/cm2 after 3600 mg extract with marginal statistical significance (p = 0.03 by one sided paired t-test). There was no correlation between total hypericin plasma concentrations and photosensitivity. In the multiple dose part, 50 volunteers received 600 mg hypericum extract t.i.d. with a daily dose of 5.6 mg of total hypericin. Comparison of UV light sensitivity before dosing with day 15 of treatment showed a slightly increased SSI sensitivity expressed by decrease of the MED from 0.17 to 0.16 J/cm2 (p = 0.005 by Wilcoxon test), and similarly, sensitivity to UV-A light increased (the mean tanning dose decreased from 9.9 to 7.8 J/cm2, p < 0.0001). This increase in cutaneous light sensitivity could be compensated by reducing irradiation time by 21%. Doses used in this study were higher than typical doses in current commercial preparations. In spite of these high doses in the double blind single dose part, frequency of side effects was equal to placebo medication and UV light sensitivity was not or only marginally increased. The study does not, however, exclude phototoxic reactions with doses above 11.25 mg of total hypericin and plasma levels above 100 micrograms/L. Furthermore, phototoxicity may be different after application of pure hypericin, since some constituents in the plant extract may exhibit protective effects. PMID- 9342770 TI - In vitro receptor binding and enzyme inhibition by Hypericum perforatum extract. AB - Hypericum perforatum L. Hypericaceae (St. John's wort), has been used since the time of ancient Greece for its many medicinal properties. Modern usage is still quite diverse and includes wound healing, kidney and lung ailments, insomnia and depression. This plant has been known to contain a red pigment, hypericin, and similar compounds, which have been assumed to be the primary active constituent(s) in this plant genus. A crude Hypericum extract was tested in a battery of 39 in vitro receptor assays, and two enzyme assays. A sample of pure hypericin was also tested. Hypericin had affinity only for NMDA receptors while the crude extract had significant receptor affinity for adenosine (nonspecific), GABAA, GABAB, benzodiazepine, inositol triphosphate, and monoamine oxidase (MAO) A and B. With the exception of GABAA and GABAB, the concentrations of Hypericum exact required for these in vitro activities are unlikely to be attained after oral administration in whole animals or humans. These data are consistent with recent pharmacologic evidence suggesting that other constituents of this plant may be of greater importance for the reported psychotherapeutic activity. Alternative pharmacologic mechanisms for Hypericum's antidepressant activity are critically reviewed and the possible importance of GABA receptor binding in the pharmacology of Hypericum is highlighted. Some of these results have been previously reported. PMID- 9342769 TI - Effects of hypericum extract (LI 160) in biochemical models of antidepressant activity. AB - Since the mechanism of the antidepressant activity of hypericum extract is not yet understood, we tested possible effects of standardized hypericum extract (LI 160) in several biochemical models relevant for the mechanism of action of other antidepressant drugs. While LI 160 was only a weak inhibitor of MAO-A and MAO-B activity, it inhibited the synaptosomal uptake of serotonin, dopamine and norepinephrine with about equal affinity-2 micrograms/ml). Moreover, subchronic treatment of rats with hypericum extract led to a significant down-regulation of beta-receptors and to a significant up-regulation of 5-HT2-receptors in the frontal cortex. The data might suggest that hypericum extract acts via similar biochemical mechanisms to other antidepressants (e.g. tricyclics). PMID- 9342771 TI - Effects of long-term administration of hypericum extracts on the affinity and density of the central serotonergic 5-HT1 A and 5-HT2 A receptors. AB - Extracts of St. John's wort, Hypericum perforatum L. (Hypericaceae), are used as a phytotherapeutic antidepressant. A number of clinical studies demonstrate that their antidepressive potency is comparable to tricyclic antidepressants (TCA). Although the therapeutic effect of hypericum extracts is well documented, very little is known about the molecular mode of action. As the improvement of the depressive symptoms with both TCA and hypericum extracts only occurs significantly after a lag phase of 10 to 14 days, it is assumed that the medication causes long-term adaptations within the central nervous system. In this context, serotonergic (5-HT) receptors are of special interest. Therefore, we investigated possible alterations in affinity and density of 5-HT1 A and 5-HT2 A receptors caused by long-term treatment of rats with St. John's wort. The brain without cerebellum and brain stem of rats, treated daily for 26 weeks with a commercially available hypericum extract (2700 mg/kg LI 160) were used for membrane preparations. Affinity (KD) and amount (Bmax) of serotonergic receptors were determined by employing receptor binding assays using 3 H-8-OH-DPAT and 3H Ketanserin as selective radioligands for the 5-HT1 A and the 5-HT2 A receptors, respectively. We found that in hypericum-treated rats the number of both 5-HT1 A and 5-HT2 A receptors were significantly increased by 50% compared to controls, whereas the affinity of both serotonergic receptors remained unaltered. The data suggest an upregulation of 5-HT1 A and 5-HT2 A receptors due to prolonged administration of hypericum extracts. These results are consistent with a modification of the expression levels of serotonergic receptors caused by synthetic antidepressants. PMID- 9342772 TI - Effects of the total extract and fractions of Hypericum perforatum in animal assays for antidepressant activity. AB - A commercially available extract of the aerial parts of Hypericum perforatum, LI 160, showed pronounced activity in selected animal bioassays. These include the forced swimming test (FST) and the tail suspension test, used to determine antidepressant activity, and tests indicating activity on the central nervous system, such as body temperature and ketamine induced sleeping time. The counteracting effects of drugs known to interfere with the central dopaminergic system strongly suggested that dopamine mediated activity is important for the activity of the extract. Dose-response experiments of the total extract and of fractions rich in flavonoids and napthodianthrones produced inverted U-shaped dose response curves. PMID- 9342773 TI - Testing the antidepressant effects of Hypericum species on animal models. AB - This paper summarizes the antidepressant effects of certain Hypericum species on animal models. Although there are many drugs in clinical use for the management of human depression, most of the antidepressant drugs have undesirable side effects, some of which may limit the daily life of patients, and therefore, more specific agents with lesser side effects are necessary as a new therapeutic modality for the rational treatment of depression. In our laboratory, we observed antidepressant activity with the alcoholic extract of H. calycinum whose effects on the central nervous system of mice are almost equal to the extract prepared from St. John's wort, H. perforatum. Other species, H. hyssopifolium ssp. elongatum var. elongatum seems to have no antidepressant activity. From these data, it can be concluded that at least some of Hypericum species may have a potential use for the treatment of depression. PMID- 9342774 TI - Biologically active and other chemical constituents of the herb of Hypericum perforatum L. AB - Phenylpropanes, flavonol derivatives, biflavones, proanthocyanidins, xanthones, phloroglucinols, some amino acids, naphthodianthrones and essential oil constituents are the natural plant products known from the crude drug of Hypericum perforatum, Hyperici herba. These compounds are discussed with respect to structural features, their concentration, biological activities and their possible contribution to the clinically demonstrated antidepressant efficacy of extracts obtained from Hyperici herba. PMID- 9342775 TI - Family planning methods: new guidance. PMID- 9342776 TI - Adenosine-dopamine interactions in the ventral striatum. Implications for the treatment of schizophrenia. AB - The ventral striatum is included in brain circuits which connect brain areas classically ascribed to the motor or to the limbic system. In fact, the ventral striatum is involved in the connection between motivationally relevant stimuli and adaptive behaviours. Dopamine neurotransmission in the ventral striatum is essential for the increase in motor activity produced by motivational, salient, stimuli, such as food or novelty or by the administration of psychostimulants. Adenosine plays a role opposite to dopamine in the striatum and adenosine agonists produce similar behavioural effects as dopamine antagonists. On the other hand, adenosine antagonists, like caffeine, produce similar effects to increased dopaminergic neurotransmission in the striatum. Specific antagonistic interactions between specific subtypes of adenosine and dopapaine receptors in the basal ganglia play an essential role in the behavioural effects of adenosine agonists and antagonists. In particular, a strong antagonistic interaction between adenosine A2A and dopamine D2 receptors seems to take place in the striopallidal GABAergic neurons which originate in the ventral striatum. Therefore, adenosine A(ZA) agonists provide a potential new treatment for schizophrenia, since the dopamine D2 receptors of the ventral striopallidal neurons appear to be involved in the antipsychotic effects of neuroleptics. In fact, in animal models, the adenosine A2A agonist CGS 21680 has a profile of antipsychotic with a low liability to induce extrapyramidal side effects. PMID- 9342777 TI - Hypnotic self administration: forced-choice versus single-choice. AB - Twenty-four men and women with insomnia, age 21-50 years, self administered hypnotics under a single-choice with placebo, single-choice with triazolam (0.25 mg), or forced-choice of placebo versus triazolam (0.25 mg) paradigm. Subjects received 4- sampling nights of placebo or triazolam in the single-choice conditions or 2 nights of each in the forced-choice condition. Then on 7 choice nights they could self administer a capsule, or not, in the single-choice conditions, or were required to choose one of two color-coded capsules in the forced-choice condition. In the single-choice conditions, subjects chose placebo 80% of nights and triazolam 77% of nights, while in the forced-choice condition triazolam was chosen on 86% of nights. Thus, the self administration of triazolam did not vary significantly between single or forced choice conditions, but that of placebo did. Placebo rate was high when it was the only alternative, but low when competing with triazolam. On sampling nights, compared to placebo, triazolam produced a significant increase in total sleep time, a reduction in latency to sleep, wake after sleep onset, and percentage stage 1 sleep. Triazolam, relative to placebo, also improved mood in the morning on some sampling nights. For subjects choosing capsules < 100% of opportunities (n = 14), on nights a capsule was chosen versus nights none was chosen (regardless of whether placebo or triazolam was the choice), self-ratings 30 min before bedtime on the Profile of Mood States vigor scale were significantly higher. PMID- 9342778 TI - Social interaction: responses to chlordiazepoxide and the loss of isolation reared effects with paired-housing. AB - This study investigated the action of chlordiazepoxide (CDP), on the social interaction (SI) of adult rats maintained in one of three housing conditions: (i) group-reared, (ii) isolated from weaning or (iii) paired during adulthood after initial isolation at weaning (pair-housed former isolates; PHFIs). Group-reared rats and PHFIs rats were housed in pairs starting 21 days prior to the first experiment. For these two groups, CDP (2.5 and 5.0 mg/kg) increased SI in unfamiliar, but not familiar (cagemate) pairings. In rats isolated throughout, SI was markedly increased and this was unaffected by CDP. Isolated rats also exhibited increased motor activity (MA) during SI tests, and the MA of both isolates and PHFIs was reduced by CDP. Finally, levels of aggression were very low except in isolates, where a relatively modest increase in aggression was reversed by either pairing or CDP. To summarise, isolation-induced increases in SI and aggression were reversed by pairing, but pairing only attenuated isolation induced increases in MA. Although CDP reduced the elevated aggression and MA of isolated rats, it had no effect on their elevated SI scores. These data question the permanence of anxiety-related, isolation-induced behavioural changes and stand in contrast to the irreversible anxiogenic profile reported for isolation reared rats in the elevated X-test. PMID- 9342779 TI - Characterization of the ethanol-like discriminative stimulus effects of 5-HT receptor agonists as a function of ethanol training dose. AB - A drug discrimination procedure was used to characterize the ethanol-like effects of a variety of 5-HT1 agonists. Previous studies found that the degree of substitution of the 5-HT1B/2C agonist TFMPP (m-trifluoromethylphenylpiperazine) depended on the training dose of ethanol. The present studies extend this initial finding to four additional 5-HT agonists with different selectivity for 5-HT1A, 5 HT1B, or 5-HT2C receptors: CGS 12066B (7-trifluoromethyl-4(4-methyl-1 piperazinyl)-pyrrolo[1,2a]quinoxaline maleate), mCPP [1-(3 chlorophenyl)piperazine diHCl], RU 24969 [5-methoxy-3(1,2,3,4-tetrahydro-4 pyridinyl]-1H-indole succinate and 8-OH DPAT [(+/-)-8-hydroxy-2-(di-n propylamino)tetralin HBr]. Separate groups of rats were trained to discriminate 1.0 g/kg (n = 7), 1.5 g/kg (n = 6) or 2.0 g/kg (n = 8) ethanol from water. Following training, three to five doses of each 5-HT agonist were tested twice in each rat. The most selective 5-HT1B agonist tested, CGS 12066B (3-17 mg/kg; IP), completely substituted for the 1.0 g/kg ethanol, but not for 1.5 or 2.0 g/kg ethanol. Likewise, the 5-HT1B/2C agonist mCPP (0.56-1.7 mg/kg; IP) completely substituted only in the 1.0 g/kg ethanol training group. The 5-HT1A/1B agonist RU 24969 (0.1-3.0 mg/kg; IP) substituted for all training doses of ethanol, although in a lower proportion of the rats tested in the 2.0 g/kg ethanol training group. Finally, the 5-HT1A agonist 8-OH DPAT (0.1-1.0 mg/kg, IP) did not substitute completely for any ethanol training dose. The results consistently show that agonists with 5-HT1B activity produce discriminative stimulus effects similar to low and intermediate, but not high, ethanol training doses. PMID- 9342780 TI - The effects of kainic acid lesions on dopaminergic responses to haloperidol and clozapine. AB - The antipsychotic drugs haloperidol and clozapine have the common action of increasing dopamine metabolism in the striatum (nucleus accumbens, caudate putamen) of the rat. Intracerebroventricular administration of kainic acid (KA) produces neuronal loss in limbic-cortical brain regions which project directly or indirectly to the striatum. In the present study, dopamine metabolism in subregions of the striatum was examined in rats with KA lesions after acute and chronic haloperidol or clozapine administration. The main findings was that the elevating effect of acute haloperidol treatment on the dopamine metabolite, DOPAC, was blocked in the nucleus accumbens shell and diminished in medial and laterodorsal caudate-putamen of the KA-lesioned rats. In addition, the elevating effects of both acute and chronic haloperidol treatment on dopamine turnover were attenuated in the laterodorsal caudate-putamen of KA-lesioned rats. The levels of dopamine, DOPAC, and HVA after chronic clozapine treatment were greater in KA lesioned than control rats. These results indicate that dopaminergic responses to haloperidol may be diminished by limbic-cortical neuropathology, while such pathology does not significantly alter dopaminergic responses to clozapine. PMID- 9342781 TI - Training-dependent biphasic effects of corticosterone in memory formation for a passive avoidance task in chicks. AB - This study investigates the functional role of corticosterone in memory formation for a passive avoidance task in the day-old chick. Whereas training chicks with a strong aversant results in an enduring memory, memory for a weak aversant which is only retained for a few hours (< 9 h) is facilitated by intracerebral corticosterone administration. We now report that only chicks trained on the strong task, a learning situation that results in a high percentage of chicks (around 80%) forming a long-term memory, showed increased post-training plasma corticosterone levels, whereas chicks trained on the weak task showed corticosterone values comparable to untrained chicks. We also questioned whether the effects of corticosterone on retention might be concentration dependent. In the weak task, intracerebral administration of 1 microgram corticosterone facilitated retention, but a higher dose failed to induce this effect. However, in the strong task, corticosterone administration at doses of 1 and 5 micrograms produced an impairment in long-term retention for the avoidance response. These results support a crucial role of corticosterone release following training on the physiological mechanisms ensuring the transition from short- to long-term memory, and provide an animal learning model for the study of the mechanisms of action of a biphasic modulation of memory formation by acute corticosterone administration. PMID- 9342782 TI - A profile of the behavioral changes produced by facilitation of AMPA-type glutamate receptors. AB - A newly developed group of benzoylpiperidine drugs that enhance AMPA-receptor gated currents ("ampakines") has been shown to improve memory encoding in rats across a variety of experimental paradigms. The present experiments were intended to i) provide a partial profile of the behavioral changes produced by ampakines, ii) test if two ampakines (BDP-12 and BDP-20) that differ significantly in their effects on AMPA receptor kinetics produce similar behavioral profiles, and iii) determine if physiological potency is reflected in behavioral potency. BDP-20 reduced two measures of exploratory activity in aged rats but increased speed of performance in a radial maze; the drug also caused substantially improved retention of spatial information. These results are similar to those obtained with BDP-12, an analog that differs from BDP-20 in its effects on ligand binding to the AMPA receptor and on the physiological responses of the receptors to glutamate. BDP-20 was approximately ten-fold more potent in behavioral effects than BDP-12, which agrees with the relative potencies of the two drugs as assessed with excised patches and excitatory synaptic responses. These findings indicate that ampakines, though differing in their effects on AMPA-receptor mediated responses, have similar effects at the behavioral level. PMID- 9342783 TI - Hypnotic and hypothermic action of daytime-administered melatonin. AB - Six healthy male volunteers (average age = 22.5 years) received orally melatonin (MLT; 3 mg or 9 mg) or placebo at 0930 hours in a randomized, single-blind, cross over study. Both doses of exogenously administered melatonin (ex-MLT) induced transient, significant suppression of core body temperature (BT) compared to the placebo condition. There was no significant difference in the degree of BT suppression among the two MLT conditions in spite of significantly higher levels of serum MLT with the close of 9 mg, suggesting that there may be a threshold level of ex-MLT inducing the hypothermic action. Daytime administered ex-MLT also induced a significant sleep-inducing effect only in the 9 mg condition, while 3 mg ex-MLT failed to produce statistical significance. These findings suggest that the sleep-inducing action of ex-MLT occurs only at relatively high doses, and this action is probably not due to its BT lowering action. The present study led us to assume that ex-MLT produce its therapeutic effect for circadian rhythm sleep disorders through induction of circadian phase-shifting preceded by an acute, transient hypothermic action rather than light entrainment after setting sleep time by induction of sleep propensity. PMID- 9342784 TI - The reduction in alcohol intake by the 5-HT1A agonist 8-OH DPAT and its attenuation by the alpha 2 adrenergic antagonist idazoxan correlates with blood glucose levels. AB - Serotonergic agents in general and the 5-HT1A agonist 8-OH DPAT in particular, reduce alcohol intake in rats and primates but the mechanism of this effect is not known. Previous studies have shown a correlation between alcohol consumption and the propensity to consume sweet substances. Indeed, certain biochemical events accompanying glucose utilization have been proposed as satiety signals in the control of feeding. Since 8-OH DPAT produces hyperglycemia, we tested the hypothesis that its effect on alcohol intake may be partly mediated through an increase blood glucose. Male Wistar rats were trained to drink a bout of 6% (w/v) alcohol using the limited access procedure which offers a daily 40-min access to alcohol and water. On consecutive test trial days separated by intervening non drug days, the amount of alcohol consumed (1 g/kg on intervening days) was measured following the administration of 8-OH DPAT (150 micrograms/kg 10 min prior to drinking) alone or in combination with the prior (20 min) injection of idazoxan (2 mg/kg), an alpha-2 adrenoceptor antagonist with hypoglycemic properties. Idazoxan attenuated the hyperglycemic effect of 8-OH DPAT and completely reversed 8-OH DPAT's inhibitory effect on alcohol intake. Idazoxan alone produced a mild hypoglycemia and stimulated alcohol intake. These results support a role for glucoregulatory processes in serotonergically-mediated changes in alcohol consumption. PMID- 9342785 TI - Cross-sensitisation with d-amphetamine following repeated intra-perifornical sulpiride infusions. AB - Infusions of the dopamine D2/D3 receptor antagonist sulpiride within the perifornical region of the lateral hypothalamus have been reported to increase locomotor activity. The current investigation examined the effect of repeated lateral hypothalamic sulpiride infusions. In experiment la, rats were placed repeatedly in an activity chamber either prior to, or following an infusion of 10 micrograms sulpiride or vehicle. Repeated infusions of sulpiride prior to, but not following exposure to the activity chamber increased locomotor activity during subsequent sessions. In experiment 1b, repeated pretreatment with intra perifornical sulpiride prior to placement within the activity chamber was found to engender a significant increase in conditioned activity when placed subsequently within the same chamber drug-free. Alternatively, pretreatment with sulpiride in the home cage was found subsequently to engender a significant increase in locomotor activity during a test session with intra-perifornical sulpiride. In experiment 2, repeated pretreatment with intra-perifornical sulpiride significantly increased the locomotor response to a subsequent systemic challenge with d-amphetamine. Animals pretreated in the home cage exhibited a moderate increase in activity over vehicle controls, while animals repeatedly pretreated immediately prior to placement in the activity chamber exhibited the largest response subsequently to d-amphetamine of any group. Experiment 3 showed that repeated sulpiride infusions either 1 mm anterior or 1 mm posterior to the perifornical region were without effect upon locomotor activity. These data are suggested to reflect an indirect action of intra-perifornical sulpiride upon the mesoaccumbens dopamine projection, via the level of the ventral tegmental area. Precise neural mechanisms are under current investigation. PMID- 9342786 TI - The NMDA antagonist, dizocilpine, enhances cocaine reinforcement without influencing mesoaccumbens dopamine transmission. AB - In order to assess the role of excitatory amino acids (EAAs) in the reinforcing property of cocaine, the NMDA antagonist, dizocilpine, or the AMPA antagonist, DNQX, were administered to animals previously trained to self-administer cocaine (1.0, 0.4 or 0.16 mg/kg per infusion). The highest doses of dizocilpine (0.1 mg/kg, IP) and DNQX (30 mg/kg, IP) significantly reduced operant responding for cocaine maintained on a fixed ratio schedule of reinforcement. However, whereas dizocilpine had no influence operant responding for food, DNQX significantly decreased lever pressing for this reinforcer. These results indicate that an NMDA antagonist produces a relatively selective enhancement of cocaine reinforcement, while an AMPA antagonist decreases cocaine self-administration only at a dose that also impairs responding for an alternate reinforcer. A parallel in vivo microdialysis study performed in freely moving rats tested the effects of dizocilpine alone and in combination with cocaine on extracellular dopamine in the nucleus accumbens shell. The results revealed that dizocilpine alone (0.1 mg/kg, IP) did not alter basal extracellular dopamine levels in the nucleus accumbens or spontaneous behavior. In addition, 0.1 mg/kg dizocilpine did not alter the increase in dopamine in the accumbens shell or behavioral hyperactivity produced by cocaine (15 or 30 mg/kg). Collectively, these findings suggest that dizocilpine enhances the reinforcing effect of cocaine without influencing dopamine transmission in the nucleus accumbens shell. PMID- 9342787 TI - PET-characterization of [carbonyl-11C]WAY-100635 binding to 5-HT1A receptors in the primate brain. AB - [carbonyl-11C]WAY-100635 is a new radioligand which can be used with positron emission tomography (PET) to provide high contrast delineation of human brain regions that are rich in 5-HT1A receptors. In the present PET study, the binding of [carbonyl-11C]WAY-100635 was characterized in the cynomolgus monkey brain. Pretreatment with each of the two reference compounds, WAY-100635 and 8-OH-DPAT, as well as the drugs buspirone and pindolo, induced a marked inhibition of [carbonyl-11C]WAY-100635 binding in the neocortex and the raphe nuclei. A preliminary Scatchard analysis yielded 5-HT1A receptor density values of the same order as those that have been reported in vitro. The study shows that [carbonyl 11C]WAY-100635 binds specifically to 5-HT1A receptors in the primate brain and has potential for determination of 5-HT1A receptor occupancy and density in psychiatric patients. PMID- 9342788 TI - THA disrupts mismatch negativity in Alzheimer disease. AB - The present study investigates the effects of 25 or 50 mg tetrahydroaminoacridine (THA), a cholinesterase inhibitor, on auditory mismatch negativity (MMN) event related response in 19 patients with Alzheimer disease (AD). MMN is produced by the deviant tones representing passive attention and automatic detection of stimulus change. In an inattentive task condition, standard (85%) and deviant (15%) tones were represented in a random order with interstimulus interval of 1 s in separate blocks. THA 25 mg had no effect on MMN in AD patients. In contrast, THA 50 mg diminished MMN in AD subjects. The results suggests that acute treatment of AD subjects with a cholinesterase inhibitor disrupts the passive detection of change in an auditory input. PMID- 9342790 TI - Insulin and/or dexamethasone regulation of lactate production and its relationship to glucose utilization by ovine and bovine adipose tissue explants incubated for 7 days. AB - This study investigated the hormonal regulation of lactate production and the lactate production/glucose utilization ratio in sheep and cow adipose tissue (AT) explants. The effects of insulin (2 mU/mL) and/or dexamethasone (DEX, 100 nM) were measured in perirenal AT of adult non-lactating non-pregnant cows (n = 5) or ewes (n = 5) given a restricted diet followed by overfeeding. The AT explants were incubated for 7 days. Under basal conditions, the lactate production was two times greater in sheep than in cow AT. Insulin increased lactate production in the two species, whereas DEX decreased it. DEX, in the presence of insulin, increased lactate production from day 3 to day 7. The lactate production/glucose utilization ratio was about two times greater in sheep than in cow AT. The presence of insulin increased, and that of DEX decreased this ratio in sheep AT, whereas it was not changed in cow AT. DEX, in the presence of insulin, increased this ratio in the two species after 4 or 5 days. PMID- 9342789 TI - Collection of pancreatic juice in experimental animals: mini-review of materials and methods. AB - This article briefly describes some anatomical details of the pancreatic duct system in dogs, cats, cattle, sheep, goats, pigs and rats that are important for the preparation of pancreatic ducts for surgery. The advantages and disadvantages of various materials used for preparing cannulas and catheters for the collection of pancreatic juice are also presented. Several techniques of pancreatic duct cannulation (Thomas', duodenal pouch and Routley's methods) and pancreatic juice collection are discussed with regards to the specificity of different animal species. The results of various collections of pancreatic juice obtained in different laboratories, and resulting from the application of specific methods for particular experimental purposes are discussed. PMID- 9342791 TI - Influence of jejunal nutrients on transpyloric flow and pyloric resistance in pigs. AB - The effects of small intestinal infusion of nutrients on the transpyloric flow and pyloric resistance were evaluated in anaesthetized pigs. Saline versus isocaloric solutions of dextrose, triglycerides and casein were infused into a jejunal loop during saline gastric loading. Antropyloroduodenal pressures were measured with a sleeve/side-hole manometric assembly and the transpyloric flow with an electromagnetic flowmeter probe. Fundic pressure was maintained constant. Although the overall gastric emptying rate was not affected by nutrients, the stroke volume of the transpyloric flow pulses was significantly increased as a consequence of larger peak flow (dextrose) or longer duration of flow pulses (triglycerides and casein). Pyloric resistance was reduced by all nutrients owing to a change in the temporal relationship between the onset of pyloric pressure events and flow pulses so that flow pulses occurred after pyloric pressure events. In conclusion, under controlled fundic pressure, nutrient infusions decrease pyloric resistance. PMID- 9342792 TI - Retention time of feed particles and liquids in the stomachs and intestines of dairy cows. Direct measurement and calculations based on faecal collection. AB - To validate a method for analysing indigestible marker excretion patterns in terms of digesta passage, the mean retention time (MRT) of long hay, ground hay and concentrate, marked, respectively, with thulium, ytterbium and dysprosium was measured in the total digestive tract (TMRT) and in the stomachs (SMRT) of four cows fed on a diet of hay in the long form (17.7 kg DM/day). The MRT of the particulate and liquid phases in the intestines was obtained after faecal particles labelled by Europium and Chromium EDTA were pulse dosed through the duodenal cannula. Following test meals, total faecal collection and spot sampling of duodenal digesta were performed at fixed intervals. TMRT were 51.7, 45.6, 40.6 h and SMRT were 39.5, 31.9 and 28.0 h, respectively, for hay, ground hay and concentrate. The MRT of the liquids in the rumen (8.7 h) was shorter than the SMRT of particles but there was no differential passage between liquids and particles after the duodenum. Intestinal MRT averaged 11 h and was partitioned into 7.5 and 3.5, respectively, for MRT in the tubular sections and the caecum proximal colon. The compartmental analysis of the faecal patterns of markers given during a test meal gives the following results. The time associated with the descending part of faecal kinetics (respectively, 25.3 and 22.9 h for hay and concentrate) is directly related to the escape of feed particles from the rumen. The delay of first appearance of markers mostly reflects transit in the post duodenal tubular sections for the concentrate. The time associated with the ascending part (respectively, 16.9 and 9.4 h for hay and concentrate) represents the time required to reduce the size of the forage particles (7 h according to the difference between TMRT of long and ground hay direct measurements) and caecal mixing (3.5 h) as well as other compartments or processes that are not clearly identified. PMID- 9342793 TI - Comparison of vitamin C bioavailability after multiple or single oral dosing of different formulations in sheep. AB - The purpose of this study was to compare the bioavailability of either multiple or single oral supplementation of different formulations of vitamin C and intra duodenal supplementation of one form of vitamin C in sheep. Formulations used in the study were (1) ascorbic acid fine powder (AA); (2) ascorbic acid coated with ethyl cellulose (EC); (3) Rovimix STAY-C (SC); (4) sodium ascorbate (SA); (5) Rovimix C (RC). The bioavailability of vitamin C formulations was assessed by the changes in plasma ascorbic acid concentrations, area under the curve (AUC) and area under the curve above its basal concentration (AUCabove) values. There was no effect of single oral supplementation on bioavailability of vitamin C. Multiple dosing over a period of 28 days of oral supplementation of all five formulations resulted in higher AUCabove values. Furthermore, multiple oral supplementation of RC increased plasma concentrations of ascorbic acids and AUC values. Single intra-duodenal supplementation of ascorbic acid resulted in significantly higher AUC when compared with oral supplementation of the same vitamin C. PMID- 9342794 TI - Proximal gastric distension modifies ingestion rate in pigs. AB - Control of food ingestion related to proximal gastric distension has never been demonstrated in pigs. The aim of this study was to demonstrate its existence. Meal duration, food intake rate and characteristics of non-ingestion periods were evaluated during the ingestion of a 500 g meal with simultaneous balloon distension of the proximal stomach. Distensions were performed either at fixed pressure levels or at fixed volume levels. Five pressure levels and five volume levels were tested in duplicate experiments in random order and on different days in each animal. Pressures equal or above 11 mmHg increased meal duration (656 +/- 12 vs 562 +/- 30 s, 11 mmHg vs control) because of a lower rate of food intake and longer period of non-ingestion. On the contrary, irrespective of the gastric bag volume, isovolumic distensions did not alter feeding behaviour. We concluded that a short term control of food intake exists in pigs. PMID- 9342796 TI - Induced differentiation of ovine foetal gonocytes after grafting in the scrotum of nude mice. AB - This work was designed to elucidate, in foetal ovine testis, whether the reason why gonocytes do not differentiate into spermatogonia and do not initiate spermatogenesis is due to inadequate foetal environment (temperature and/or hormonal balance) or if somatic testicular cell proliferation is a prerequisite for initiation of male meiosis. The development and differentiation of gonocytes were analysed by grafting 60-day-old foetal ovine tests into the scrotum of immunotolerant adult Nude mice. Forty days after grafting, nine of the ten grafted testes had survived but had not increased in weight as compared to 60-day old tests. Moreover, only one third of the graft was occupied by testicular tissue, in which the relative proportions of intertubular tissue and sex cords were not altered when compared with those of normal foetal testes. The remainder of the graft was occupied by teratoma. The total number of Leydig (-80%) cells, Sertoli (-66%) cells, gonocytes (-90%) and the total length of sex cords (-63%) per grafted testis were always significantly reduced (P < 0.02), whereas the sex cords were significantly increased in diameter (+36%; P = 0.02) as compared to those of non-grafted 60-day-old foetuses. However, in seven out of the nine testes, type A spermatogonia were obtained and in two of the seven a few type B or leptotene primary spermatocytes could be observed. The grafting of foetal testis in an adult scrotum induces differentiation of gonocytes into spermatogonia, independently of proliferation of Sertoli and Leydig cells. PMID- 9342795 TI - Effect of immunisation against leukaemia inhibitory factor on the establishment of pregnancy in sheep. AB - Leukaemia inhibitory factor (LIF), a pleiotropic cytokine, is implicated in blastocyst implantation in mice and maintains the development of ovine embryos in culture. Previously, LIF mRNA and protein were demonstrated in the endometrium throughout the oestrous cycle and early pregnancy in the ewe. In this study pregnant ewes were passively immunised against human recombinant LIF with polyclonal antibodies raised in cows by active immunisation. Ewes were immunised during two stages of early pregnancy: blastocyst development to hatching, and blastocyst elongation to implantation. Only animals passively immunised against LIF showed detectable LIF antibodies in their sera and in uterine lumina flushings by radioimmunoassay and Western blot analysis. Pregnancy was confirmed by ultrasound on day 55 and a 33.5% non-significant decrease in pregnancy rate of anti-LIF treated animals was observed, when compared to animals in control groups (untreated or treated with bovine anti-keyhole limpet hemocyanin). Cows actively immunised with recombinant human LIF and exhibiting high levels of LIF antibodies in their sera at the time of blastocyst implantation also showed a reduced pregnancy rate in comparison to control animals. Although LIF may not be obligatory for implantation in ruminants it does appear to have a role during the establishment of pregnancy. PMID- 9342797 TI - Brain, breathing and breathlessness. AB - This review attempts to summarize: (i) evidence on how man voluntarily or behaviourally (as in speech) alters breathing; and (ii) evidence on how the breathlessness induced by CO2 inhalation, is perceived. The application of new methods to study these problems, e.g. functional brain imaging and transcranial focal brain stimulation, is summarized. Studies of patients with specific neurological lesions have shed considerable light in this area. The key requirement for the ponto-medullary respiratory oscillator to be both 'intact' and 'responsive' for the perception of CO2-induced air hunger is emphasized. We are ignorant as to how the voluntary/behavioural control system interacts with the automatic system at any site above the final common pathway of the respiratory anterior horn cells in the cervical and thoracic spinal cord. The opportunities for further work are outlined. PMID- 9342798 TI - The effect of the level of ventilatory assist on the level of respiratory drive in decerebrate cats. AB - The present study was undertaken to investigate whether, independent of changes in PaCO2, ventilatory assist influences not only the pattern but also the level of the respiratory drive. The experiments were performed on decerebrate and paralyzed cats ventilated by a phrenic-driven servo respirator at three different FICO2 levels (0, 0.30, 0.05). The level of ventilatory assist was altered within the range where PaCO2 did not exceed 80 Torr. A higher FICO2 accompanied a higher level of ventilatory assist. The relationship between the minute phrenic activity and log10 PaCO2 at a given FICO2 was linear. No significant difference was found in the regression lines at different levels of FICO2. We conclude that ventilatory assist has little effect on the respiratory drive at a constant level of chemical feedback during hypercapnia. PMID- 9342799 TI - Modulation of the ventilatory increase at the onset of exercise in humans. AB - The fast initial increase in ventilation at the start of exercise is generally assumed to be of reflex origin (exercising limbs) and/or caused by a 'feedforward' mechanism increasing breathing via brainstem respiratory centres or cortical areas controlling respiratory muscles. We wanted to test whether this ventilatory increase is in part a learned response which can be modified. Eleven subjects did two 20 min low-intensity arm-cranking exercise bouts on eight different days. Seven subjects were assigned to the experimental group which performed exercise paired with an 1.5 L external dead space. Before and after their eight exercise 'training'-days, these subjects did the same exercise without dead space. At the beginning of the first post-training exercise test (without dead space), the ventilatory increase at the start of exercise (sum of the first four breaths) was significantly increased (31.1 +/- 4.1 L . min-1) compared to the pre-training test session (24.4 +/- 3.9 L . min-1). No significant change was observed in the control group. We conclude that part of the ventilatory increase at the start of exercise can be modulated and might possibly be a learned response. PMID- 9342800 TI - The cardiorespiratory response to anoxia: normal development and the effect of nicotine. AB - Maternal smoking increases the risk of the sudden infant death syndrome (SIDS) 2 4-fold. The mechanism is unknown but may be related to hypoxia responses. Recovery from hypoxic apnea by young mammals depends on gasping and bradycardia. We asked whether prenatal nicotine exposure, reported to reduce hypoxic survival in 2 day old rat pups, acted by impairing gasping or bradycardia. Pregnant rats were infused throughout gestation and 1 week postnatally with nicotine tartrate (NIC) 12 mg/kg per day or saline (CON). Maternal plasma nicotine was 134.4 +/- 42 ng/ml, significantly reducing pup body weight. Pups at 3-28 days were exposed to anoxia (97% N2/3% CO2) until gasping ceased, while breathing and heart rate were recorded. NIC and CON groups were not significantly different at any age, in baseline heart rate, respiratory rate, the time course for bradycardia, time to gasp onset, duration of gasping, or number of gasps, although most of these variables declined significantly with age. We conclude that responses to anoxia are not affected by prenatal high-dose nicotine. PMID- 9342801 TI - Day-night differences in the respiratory response to hypercapnia in awake adult rats. AB - We investigated whether resting ventilation and the hypercapnic ventilatory response vary with time of day in awake adult rats. Respiratory frequency (fR), tidal volume (VT), inspired ventilation (VI), inspiratory interval (tI), carbon dioxide production (VCO2) and abdominal temperature (Tb) were measured before and during a hypercapnic stimulus (3.5% CO2 in air) at 10:00 and 22:00 h. VCO2, Tb and mean inspiratory air flow (VT/tI) were significantly higher at 22:00 h in air. VI/VCO2 was similar at 10:00 and 22:00 h. VI was significantly elevated by hypercapnia and the response at 22:00 h was 2.3 times greater than that at 10:00 h. VT/tI was unchanged at 10:00 h but significantly increased by hypercapnia at 22:00 h. VCO2 was significantly depressed at 10:00 h but not at 22:00 h. Tb was unaffected by hypercapnia. We conclude that the metabolic and ventilatory responses to hypercapnia are dependent on the time of day. PMID- 9342802 TI - Hypoxemia and hypoxic pulmonary vasoconstriction: autonomic nervous system versus mixed venous PO2. AB - Hypoxemia interferes with hypoxic pulmonary vasoconstriction (HPV). We investigated the respective roles of the autonomic nervous system and the mixed venous PO2 (PVO2) in the attenuation of HPV by hypoxemia. Pentobarbital anesthetized dogs had their lungs separately ventilated with a dual-lumen endotracheal tube. Left (Ql) and total (Qt) pulmonary blood flows were determined using electromagnetic flow probes. HPV was initiated by ventilating the left lung with nitrogen for 5-10 min while the right lung received 100% oxygen. The animals were subsequently made hypoxemic by switching the right lung to room air ventilation (5-10 min). Two different protocol groups received either intravenous atropine during hypoxemia (group I) or intravenous propranolol prior to protocol initiation (group II). A third group of dogs (group III) had their mixed venous PO2S maintained above 30 torr during hypoxemia. In response to left lung hypoxia, Ql/Qt decreased from 44 +/- 5, 48 +/- 3 and 46 +/- 2% to 25 +/- 4, 28 +/- 2 and 26 +/- 3% in the three groups, respectively. During hypoxemia Ql/Qt increased to 50 +/- 7 and 47 +/- 3% in groups I and II. In group III dogs, Ql/Qt remained significantly decreased at 31 +/- 3%. Subsequent administration of atropine in group I had no effect on Ql/Qt. We conclude that the loss of flow diversion from a hypoxic lung during hypoxemia may be mediated primarily by a decreased in mixed venous PO2 when PVO2 is allowed to decrease to 15-20 torr. PMID- 9342803 TI - Effect of hydration on lung interstitial conductivity response to electrically charged solutions. AB - In interstitial segments of rabbit lung, we compared the flow of a solution containing cationic protamine sulfate (0.08 mg/ml) or cationic dextran (0.1%) to that of Ringer or neutral dextran solution. Also compared, were the flow of solutions containing anionic dextran (0.1 or 1.5%) to those containing neutral dextran and the flow of hyaluronidase solution (0.02%) to that of Ringer solution, at mean interstitial pressures (Pm) between -5 and 15 cmH2O. Driving pressure was set at 5 cmH2O. Cationic protamine or cationic dextran-to-Ringer flow ratio increased with Pm (presumably as hydration increased) but in nonedematous interstitium (-5 cmH2O Pm), flow ratio was 1, indicating a viscosity dependent flow. In contrast, the flow of anionic dextran solution decreased relative to that of neutral dextran; this decrease was constant with hydration, but was greater at the higher concentration of dextran. Interstitial conductivity to the flow of hyaluronidase increased with hydration. However, this behavior was absent after the flow of 1.5% anionic dextran, indicating an inhibitory effect of the higher concentration of anionic dextran on the hyaluronidase response. A negative charge in microvascular filtrate may control fluid clearance in normal interstitium, while a positive charge would enhance clearance only in edema formation. PMID- 9342805 TI - Tropical diseases: the burden and its implications. AB - The weight of disease in developing countries may be viewed as a burden, but it also provides opportunities and challenges in research, practical and altruistic action and ecological understanding. Of the classical parasitic diseases, 10 are to be found in the top 100 major diseases in the world. The burden of tropical diseases is much greater in Africa, with far fewer resources available. Infectious diseases are combined with diseases of poverty. High fertility, slow economic growth, deforestation, rapid urbanization and increased migration contribute to the recent changes in transmission and distribution of diseases observed in developing countries. The burden of diseases in tropical and developing countries remains massive. New, especially molecular, parasitological techniques and lines of research, and scientific methods for investigations and management of tropical diseases can contribute to the reduction of the burden of disease while providing a better understanding of parasites and their interactions with their hosts. PMID- 9342804 TI - Factors related to cervicitis in Qashghaee nomadic women of southern Iran. AB - In a survey of reproductive habits and cervical pathology in 1015 women of the Qashghaee nomadic tribe of southern Iran, a high prevalence (80%) of cervicitis was observed. The effects of reproductive and other habits on risk of cervicitis were therefore assessed using proportional odds modelling. Increased risks of a higher degree of cervicitis were associated with middle to late age (p = 0.02), a history of delivery of children (p = 0.001), intercourse during menses (p = 0.03) and a history of ever giving birth in a maternity hospital (p = 0.02). A lower risk was associated with increased duration of marriage (p = 0.05). The increased risk observed for history of birth in maternity hospital may reflect a prior obstetric problem which predisposes to both cervicitis and seeking the specialist obstetric care provided in the maternity hospital. Results suggest an infectious agent which may be transmitted in childbirth. PMID- 9342806 TI - Veterinary parasitology and human health. AB - Veterinary parasitology is a subdiscipline of veterinary medicine and is concerned (a) with diseases in animals caused by protozoan and metazoan parasites, (b) with parasitic zoonoses and (c) with parasites acting as disease vectors. Veterinary parasitology is related to human health and well-being in a number of ways. It plays a role in treatment, prevention and control of parasitoses in animals which serve as food sources for humans, which are used in agriculture as farm animals or which are companion animals of humans. Of direct significance to human health are activities of veterinary parasitology in prevention and control of food-, water-, vector-borne and other zoonoses, such as toxoplasmosis, cryptosporidiosis, leishmaniosis, taeniosis/cysticercosis, echinococcosis and others. In this context the evolution of veterinary parasitology, progress in the control of parasitic animal diseases, the concerns of mass-treatment of livestock against parasitoses, environment related problems and other factors are discussed. Finally, examples of current and predictable future research trends in veterinary parasitology are summarized. It is concluded that an innovative veterinary parasitology in cooperation with other disciplines will be in a position to contribute further to the improvement of human health and well-being. PMID- 9342807 TI - The future of tropical medicine in Europe. Opportunities for intersectoral cooperation. AB - The author provides a brief history of tropical medicine and describes how it has evolved since its creation at the end of the 19th century. The author proposes a working definition of tropical diseases, including their geographical distribution and their socioeconomic conditions prevailing in these regions. Currently tropical medicine is in danger of fragmenting into a number of subspecialties or being absorbed into the subspecialties of other medical sectors. The broad interdisciplinary nature of tropical medicine may be its weakness, but also its particular strength. The effective interplay between four major areas of work makes it different from other specialties. These are biomedical sciences, medical care, epidemiology and the social sciences. This intersectoral approach should also lead to stronger synergisms between scientists in Europe and institutions in the South. PMID- 9342808 TI - Scientific research partnership: north-south and south-south. AB - Scientific research can contribute much to solving many urgent global problems. However, research must be conducted in a global context and, in particular, must be extended to the countries of the South. If this is to be done, the research capacities of those countries must be considerably expanded and consolidated. Swiss strategy for promotion of research in the developing world is primarily devoted to this aim. Problems of general concern must be solved by joint efforts, which means in development-oriented, long-term, interdisciplinary research partnerships. In this context the requisite training of everybody involved can be conducted in a continuous way. Partnership is a severely taxing activity. In six points it is shown that today the greater portion of the Swiss research community does not yet meet these requirements. Despite relatively scarce funds, the time has come to review the Swiss university world in this respect also. PMID- 9342809 TI - Avascular osteonecrosis of the acetabulum. AB - OBJECTIVE: To investigate the possible occurrence of osteonecrosis in the acetabulum in patients with non-traumatic necrosis of the femoral head. DESIGN AND PATIENTS: One hundred and seventy-nine patients with non-traumatic femoral head necrosis were assessed by MRI and radiography for the presence of acetabular necrosis. Three criteria were established to differentiate between osteonecrosis and osteoarthritic changes: (1) heterogeneous morphology and irregular contours of the lesion; (2) typical demarcation lines of osteonecrosis; (3) deficient accumulation of intravenous gadolinium in the affected regions. RESULTS: In four patients histological confirmation of acetabular necrosis was obtained. The MR analysis of 22 acetabula (9.5% of those examined) showed changes which suggested osteonecrosis. No cystic lesions were demonstrated in the subchondral bone of any patient. Two cases of acetabular necrosis were found without an ipsilateral femoral head necrosis. In two patients of the 14 who had undergone total hip replacement following necrosis of the femoral head, aseptic loosening of the acetabular component was found. CONCLUSION: The study suggests that acetabular necrosis may be an accompaniment to aseptic necrosis of the femoral head. Further work is required to assess its importance in premature loosening of the acetabular element of total hip arthroplasty. PMID- 9342810 TI - Role of MRI in the diagnosis of insufficiency fractures of the sacrum and acetabular roof. AB - OBJECTIVE: To review the risk factors and the radiological appearance of insufficiency fractures of the sacrum and acetabular roof. DESIGN AND PATIENTS: Twenty patients with sacral and acetabular roof insufficiency fractures were reviewed retrospectively. There were 16 women (80%) and 4 males (age range 48-86 years, excluding an 8-year-old boy). Thirteen patients had a known tumour, and nine had received pelvic irradiation. All patients, except one who was asymptomatic, presented with low back or hip pain. In patients with a known tumor, metastases were suspected. Plain radiography (20), bone scintigrams (16), MR examinations (20), and bone densitometry (14) were performed. Nine patients also each had a CT scan. RESULTS AND CONCLUSIONS: In three cases the CT scan performed 10-25 days after onset of symptoms was interpreted as normal. MR examination performed a few days after the CT scan showed in each of these three patients a fracture line with a band of edema. Scintigraphy was very sensitive, but the H-shaped pattern of sacral uptake, specific for an insufficiency fracture, was detected in only three of 16 cases. The earliest MR sign was medullary edema, seen as early as 18 days after the onset of symptoms. On spin echo (SE) T1-weighted images (T1WI), the hypointense signal of edema could mask a fracture line. On SE T2WI the fracture line could be detected within the hyperintense edema (10 of 17 patients with examinations including SE T2WI). However, in four patients a fracture of the sacrum was not seen on T2WI, these having been obtained in the axial plane. For this reason, intravenous gadolinium was injected, revealing a fracture line in 12 of 14 examinations, or fat suppression sequences were performed, revealing a fracture line in five of five cases. The total number of fractures detected was 17 [15 fractures of the sacrum (bilateral in 10 cases) and two of the acetabular roof]. At a later stage, the edema resolved and the fracture was clearly seen. The two cases of fracture of the acetabular roof were easily recognized at MRI, particularly in the sagittal plane. PMID- 9342811 TI - MR appearance of parasymphseal insufficiency fractures of the os pubis. AB - OBJECTIVE: To clarify the MRI features of parasymphyseal insufficiency fractures of the os pubis. DESIGN AND PATIENTS: MRI was performed in four postmenopausal women with parasymphyseal insufficiency fractures. The diagnosis was confirmed with plain films in every patient. T1-weighted and T2-weighted images were obtained in four patients using a 1.5-T unit. Postcontrast T1-weighted imaging was also done in three patients. RESULTS AND CONCLUSIONS: MRI of pubic parasymphyseal insufficiency fracture characteristically demonstrates a hyperintense mass lesion with a hypointense rim on T2-weighted imaging, showing peripheral and septal enhancement after contrast administration. It is important to have this entity in mind in patients with osteoporosis, especially in patients with a history of pelvic irradiation for malignant disease, so as not to misinterpret it as a chondroid tumor or bone metastasis. PMID- 9342812 TI - Distal fibular notch: a frequent manifestation of the rheumatoid ankle. AB - OBJECTIVE: To describe the distal fibular notch, an infrequently described manifestation of rheumatoid arthritis, and to speculate on its etiology through gross dissection, histologic correlation and MR imaging. DESIGN AND PATIENTS: One hundred and twenty-one conventional ankle radiographs were obtained and reviewed in 76 patients with clinically diagnosed rheumatoid arthritis. Additional imaging of three ankles was obtained utilizing CT and MR imaging. In addition to evaluating erosive changes, note was made of the presence and location of a well defined scalloped defect along the medial border of the distal fibula. Ankle specimen dissection and histoanatomic examination was performed in an attempt to determine the exact pathogenesis of this fibular notch. RESULTS: The distal fibular notch was identified in 52 of 121 ankles (43%). Seventy-five percent of notches were syndesmotic and extended down to the horizontal ankle joint level, while 25% of notches were syndesmotic with extension below the joint. The majority of ankles (79%) demonstrated coexistent marginal erosions and/or joint narrowing. Ankle specimen dissection revealed a single-celled synovial fold within the distal tibiofibular syndesmotic recess without underlying articular cartilage extension. CONCLUSION: The fibular notch within the distal tibiofibular syndesmosis is a frequent manifestation of rheumatoid arthritis and appears to result from synovial proliferation rather than from mechanical instability. PMID- 9342813 TI - MR findings in iliotibial band syndrome. AB - OBJECTIVE: To elucidate the MR findings in iliotibial band (ITB) syndrome. DESIGN AND PATIENTS: The subjects comprised four patients (five knees) with lateral knee pain: two athletes and two non-athletes. One non-athlete was engaged in work requiring repetitive knee movement, and the other suffered from Cushing syndrome and had bilateral abnormalities. All patients were suspected of having a lateral meniscal tear prior to MR examination, but physical examination following provisional MR diagnosis warranted the final diagnosis. MR studies included fast spin echo sagittal imaging, fat-saturated fast spin echo proton density coronal imaging, and T2* radial imaging. Twelve normal volunteers were examined. RESULTS AND CONCLUSION: Fat-saturated coronal imaging demonstrated an ill-defined, high intensity area deep to the ITB. T2* radial imaging showed an identical, but less conspicuous, abnormality. The MR finding suggested soft tissue inflammation and/or edema rather than focal fluid collection in the bursae. The signal alteration predominated in the region beneath the posterior fibers of the ITB, thus supporting the current opinion that the posterior fibers of the ITB are tighter against the lateral femoral epicondyle than the anterior fibers. The ITB itself did not show any signal alteration or increased thickness. PMID- 9342814 TI - A comparison of pulse sequences in the detection of post-traumatic bone marrow abnormalities at low field strength MRI. AB - OBJECTIVE AND PATIENTS: One hundred and forty-one patients with recent joint trauma, aged 12-71 years, were imaged on a 0.2-T dedicated MRI system and evaluated for bone bruises. The most beneficial sequences were compared. DESIGN: The diagnosis of post-traumatic bone marrow abnormalities was established in 20 of 141 patients on the basis of decreased signal intensity on T1-weighted SE and GRE sequences and increased signal intensity on T2-weighted TSE and fat suppressed IRGE sequences. Signal changes within the bone marrow were evaluated and statistically correlated with normal bone. RESULTS: The highest signal alteration was found on T1-weighted SE and GRE sequences, followed by IRGE, which detected smaller differences in signal intensity. T2-weighted TSE imaging showed the least contrast. The areas with bone marrow changes were approximately equal in size on T1-weighted SE and T2-weighted TSE sequences. The same areas depicted on IRGE and GRE sequences proved to be significantly larger (P < 0.01). CONCLUSION: Using a 0.2-T dedicated system T1-weighted SE, T1-weighted GRE and IRGE sequences were most effective in detecting conspicuous bone marrow alteration, while the T2-weighted TSE sequence was inferior. GRE and IRGE imaging showed areas about 4 times larger depicting bone marrow changes. On suspicion of bone bruise, a protocol including GRE and IRGE pulse sequences could be most beneficial. PMID- 9342815 TI - The correlation between dual-energy X-ray absorptiometry measures of bone density in the proximal femur and lumbar spine of children. AB - OBJECTIVE: To assess the correlation between a pediatric patient's proximal femur and lumbar spine bone mineral density (BMD) Z-scores, and the side-to-side difference between proximal femurs. DESIGN: Three hundred and thirty-nine patients aged 2.2-17.0 years with an assortment of underlying conditions underwent dual-energy X-ray absorptiometry (DXA) measures of BMD in both proximal femurs and the lumbar spine. RESULTS: Z-scores in the proximal femur and lumbar spine correlated highly (r = 0.73, P = 0.0001), but for individual patients the difference was often significant, and increased as BMD deviated further from normal. For patients with proximal femur Z-scores of 1 to -1 the mean difference between proximal femur and lumbar spine Z-scores was 0.5; with proximal femur Z scores of less than -3 the mean difference was increased to 1.7. In conditions which symmetrically involve the lower extremities, the right and left proximal femur Z-scores differed on average by only 0.2. CONCLUSION: BMD measurements for pediatric patients are most easily interpreted by clinicians if converted to Z scores, yet these are usually available only for the lumbar spine. Age-normalized BMD assessment at more than one site is necessary to provide a more reliable, complete assessment of bone mineral status in pediatric patients. PMID- 9342816 TI - Luxatio erecta of the hip: a critical retrospective. AB - The term "luxatio erecta" has been borrowed from the shoulder to identify rare traumatic hip dislocations in which there is inferior dislocation of the femoral head and inversion of the femoral shaft. A review of the literature is presented along with an additional illustrative case. The mechanism of injury, and the radiological and physical appearance of the patient, indicate that there are two subtypes of dislocation hitherto lumped together under the single term. PMID- 9342817 TI - Medial Segond-type fracture: cortical avulsion off the medial tibial plateau associated with tears of the posterior cruciate ligament and medial meniscus. AB - We describe an unusual cortical avulsion fracture off the medial tibial plateau of the knee associated with tears of the posterior cruciate ligament and the medical meniscus. This constellation of findings is the reverse of that seen with the Segond injury complex. We postulate that the plain film diagnosis of this fracture, like the Segond fracture, is a clue to the likely presence of associated ligamental and meniscal tears, and to the mechanism of injury. PMID- 9342818 TI - Bilateral fracture dislocation of the sacroiliac joint. AB - We present a rare case of a 27-year-old man sustaining a bilateral fracture dislocation of the sacroiliac joints without disruption of the anterior pelvis, following a fall from a height. Reconstructed images in the coronal plane and three-dimensional CT images were invaluable in the diagnosis and assessment of this injury. PMID- 9342819 TI - Malignant fibrous histiocytoma at the site of a total hip replacement: review of the literature and case report. AB - Tumors developing in association with metallic implants are being reported with increasing frequency in the orthopedic literature. The authors report the tenth case of malignant fibrous histiocytoma associated with a total hip replacement. The diagnosis was made during an evaluation for rapid radiographic osteolysis surrounding the femoral component detected less than 1 year after revision of a hemiarthroplasty to a total hip replacement. The short latency period and aggressiveness of this tumor mimicking rapid osteolysis prompted a review of the literature, etiology and significance of implant-associated malignant fibrous histiocytoma. PMID- 9342820 TI - Metachronous multiple aneurysmal bone cysts. AB - We report on a 9-year-old boy with metachronous aneurysmal bone cysts involving the tibia and pubis respectively. The patient has been clinically and radiographically followed for 3 years. PMID- 9342821 TI - Mycobacterium avium infections in animals. Literature review. AB - Mycobacterium avium causes tuberculosis in chickens and other fowls but can also infect an extensive range of different animal species. The authors reviewed the available literature on this organism to show the importance of M avium infection. PMID- 9342822 TI - Salmonella typhimurium contamination from farm to meat in adult cattle. Descriptive study. AB - The aim of this work was to study the increase in hair contamination by salmonella in cattle between the farm and slaughterhouse and to explore the possible relationship between this contamination and the contamination of carcasses and of the ground beef made from these animals. Between April 1994 and May 1995, eight groups of ten cows were sampled at different stages during transportation between the farm and the slaughterhouse and on the slaughterline. For each group, one or two cows were included in each group because they had been shown to excrete Salmonella typhimurium 15 days before slaughtering. Samples were collected from the animals (faeces, hide, carcasses, lymph nodes, ears), from the environment (vehicles, cubicles, loading corridor, stunning area) and from the final product (ground beef). The hair samples as well as the environmental samples were the most frequently contaminated (26 to 69%). Eleven different salmonella serotypes were identified, with a maximum of three different serotypes per sample. The typhimurium serotype was isolated from 67% of the positive samples. For the animals leaving the farms, the frequency of hair contamination by serotype typhimurium was 8%. The step that most influenced hair contamination seemed to be the transportation to the slaughterhouse with the contamination frequency reaching 25%. The time spent by the animals in the cubicles of the waiting area of the slaughterhouse seemed to have little influence on the frequency of hair contamination. Even though the frequency of coat contamination reached 25% (for serotype typhimurium) at the beginning of the slaughterline, carcass contamination was only 1% before chilling and only involved one group of animals. In this group, hair contamination after slaughter (serotype typhimurium) reached 90% (9/10), and 80% (4/5) of the samples taken from the ground beef were positive (serotype typhimurium). No contamination was detected in the ground meat made from the other groups. PMID- 9342823 TI - Influence of stocking density on some behavioural, physiological and productivity traits of broilers. AB - In order to investigate the influence of stocking density on broiler welfare, 17,616 Ross chickens were assigned to three different treatments: T1, T2 and T3 with a final stocking density of 27, 35 and 43 kg/m2, respectively (corresponding to an initial density of 12, 16 and 20 birds/m2). Animal welfare was assessed by measuring behavioural, physiological and productivity traits. Behavioural observations included the disturbance frequency of resting birds by other birds, the duration of the lying bouts and the standing/lying ratio. The heterophil/lymphocyte ratios were assessed from blood collected before departure to the slaughterhouse. Main productivity traits were the final live weight and carcass degradation due to foot and pad dermatitis and breast blisters. Most of the observed parameters were adversely affected by the highest density (P < 0.05). Between T1 and T2, some traits tended to demonstrate that a better degree of bird welfare existed in T1 (higher standing/lying postures ratio and final live weight, lower frequency of pododermatitis and hock lesions; P < 0.05) whereas other traits showed no differences (frequency of disturbances by other birds during resting, heterophil/lymphocyte ratio). In conclusion, a stocking density of 43 kg/m2 seemed to induce poor bird welfare whereas it was not clearly demonstrated that 27 kg/m2 was better than 35 kg/m2. PMID- 9342824 TI - Survey of Babesia divergens antibody kinetics in cattle in western France. AB - Data collected from a longitudinal survey carried out over a 2-year period, in four dairy herds in western France, were used to assess Babesia divergens antibody kinetics. Farms were visited once a month. The total number of animals monitored was 236 including calves, heifers and cows of the Holstein and Normande breeds. An ELISA was used to follow the humoral response levels against Babesia divergens. When the study began, in the autumn of 1991 (200 animals), half of the animals were already seropositive (57.5%). In all four herds, the percentage of positive animals decreased during winter, and increased during spring. Antibody levels remained stable in 49 animals, high for some, very low or negative for others. Most seropositive animals showed at least one antibody peak at some time during the 2-year period, but some presented two to five. Among the calves, 61.3% showed seroconversion during the first pasture season. Similarly, 60% of the newly purchased cows showed increases in antibody levels during the 3 months after their introduction into a new herd, regardless of the initial antibody level. Only three dairy cows had expressed a clinical babesiosis, these cows were already seropositive. Clinical incidence is low in the four farms, nevertheless serological prevalence and incidence are high. PMID- 9342826 TI - Formation of flavor compounds in cheese. PMID- 9342825 TI - Morphological alterations of blood platelets induced by platelet activating factor (PAF) and partial inhibition by ketoprofen in calves. AB - The influence of platelet activating factor (PAF) was investigated in vivo on the ultrastructure of bovine platelets, and on the platelet count. The effect of an intravenous administration of ketoprofen (a non-steroidal anti-inflammatory drug) pretreatment followed by PAF infusion was also observed in a group of six healthy male Friesian calves. PAF infusion alone caused a moderate thrombocytopenia, which peaked one minute post challenge and returned to levels not significantly different from control after 30 min. Electron microscopy revealed that after PAF infusion, platelets lost their lentiform shape and became irregular, with many pseudopods. Their microtubules became impossible to distinguish. The numbers of alpha granules and dense bodies were significantly decreased. Glycogen particles became rare or even disappeared. Giant platelets occasionally appeared. The Golgi apparatus was more often visible and the number of mitochondria was significantly increased. Ketoprofen pretreatment lowered PAF-induced thrombocytopenia and decrease in the number of dense bodies. Under these conditions, the Golgi apparatus was rarely visible and giant platelets were not observed. These results showed that the morphological ultrastructure of blood platelets in bovines were modified following PAF infusion and that ketoprofen pretreatment before PAF infusion provided partial protection, limiting the extent of the morphological alterations and maintaining a normal platelet count. PMID- 9342828 TI - Gene transfer among bacteria in natural environments. PMID- 9342829 TI - Microbial production of docosahexaenoic acid (DHA, C22:6). PMID- 9342830 TI - Acetone odor and irritation thresholds obtained from acetone-exposed factory workers and from control (occupationally unexposed) subjects. AB - Sensitivity of olfaction (smell) and chemesthesis (irritation) was evaluated for 2-propanone (acetone) and 1-butanol in acetone-exposed workers (AEW; N = 32) during a workday and unexposed subjects (microES; N = 32). Irritation sensitivity was assessed using a method that relies on the ability of individuals to localize irritants on the body. When a volatile compound is inhaled into one nostril and air into the other, the stimulated side can be determined (lateralized) only after the concentration reaches a level that stimulates the trigeminal nerve (irritation); compounds stimulating olfaction alone cannot be lateralized. Intranasal lateralization thresholds offer an objective measure of sensory irritation elicited by volatile compounds. Test results indicated that neither olfactory nor lateralization thresholds for butanol differed between AEW and microES. Olfactory thresholds to acetone in AEW (855 ppm) were elevated relative to those of microES (41 ppm), as were lateralization thresholds (36,669 ppm and 15,758 ppm, respectively). Within AEW, level of occupational exposure was not correlated with thresholds. Other measures revealed that microES used more irritation descriptors than did AEW on trials where the acetone concentration was below the lateralization threshold. This is noteworthy because microES received lower concentrations of acetone to evaluate than did AEW. These results suggest that exposures to acetone induce changes in acetone sensitivity that are specific to acetone. The acetone concentrations eliciting sensory irritation using the lateralization technique were all well above current occupational exposure standards. The current study indicates that acetone is a weak sensory irritant and that sensory adaptation is an important factor affecting its overall irritancy. PMID- 9342832 TI - A sampling and analytical method for the simultaneous determination of multiple organonitrogen pesticides in air. AB - An air sampling and analytical method was developed for organonitrogen pesticides using a combined filter and XAD-2 sorbent sampler and high performance liquid chromatography-ultraviolet detection. The method was evaluated for 14 organonitrogen pesticides by National Institute for Occupational Safety and Health evaluation guidelines and procedures. Evaluation experiments addressed limits of detection and quantitation, analytical recovery, sampler capacity, sample stability, and precision and bias over a range of 12 to 240 micrograms per sample. Samples were stable when stored for up to 30 days under either ambient or refrigerated conditions. Based on the finding of this work, 10 of the 14 compounds studied (aldicarb, captan, carbaryl, carbofuran, chlorpropham, diuron, formetanate, methiocarb, oxamyl, propham) can be successfully determined simultaneously using one method with an accuracy of better than +/- 25% of the true value with 95% confidence. Two other compounds (carbendazim/benomyl, methomyl) can be measured with the same accuracy over a more limited concentration range. The remaining two compounds (propoxur, thiobencarb) may meet this criterion, but additional samples would need to be included in the data analysis. With the current data, these two compounds can be determined with an accuracy of better than +/- 27% of the true value with 95% confidence. PMID- 9342831 TI - Field testing of new aerosol sampling method with a porous curved surface as inlet. AB - A new aerosol sampling method, utilizing a porous curved surface as the sampling inlet, has recently been developed. Previous laboratory evaluations of this method have demonstrated its important features, such as low wind sensitivity and good filter collection uniformity. In this study a prototype incorporating the new method was evaluated in the field as a stationary and personal sampling device. The small sampler, utilizing a 25-mm filter is called the button aerosol sampler and was evaluated for collecting total airborne dust and fungal spores. The study was performed in nine poorly maintained inner-city houses during environmental cleanups at different cleanliness levels. The button sampler was used in parallel with the standard 37-mm closed-face filter cassette. Four collocated samplers of each type were tested at all sites as stationary samplers, and three samplers of each type were tested at two sites as personal samplers. Aerosol samples were collected on filters and analyzed using the gravimetric method for total dust and epifluorescence microscopy for fungal spores. The average particle concentration values measured with the button sampler and with the standard sampling cassette were found to correlate well within ranges of 10(1)-10(3) micrograms/m3 for total dust and 10(3)-10(5) spores/m3 for airborne fungi. The measurement results obtained with the new button sampler showed lower intersample variations of the measured concentration levels and higher uniformity of the particle deposits on the filters than those obtained with the standard cassette. PMID- 9342833 TI - Investigation and remediation of diesel converted trolley buses associated with extensive fungal growth and health complaints. AB - Fifteen bus drivers, operating diesel converted trolley buses, experienced symptoms including watery and itchy eyes, rhinorrhea, and headaches. A total of 49 buses were labeled as "problem buses" and operators refused to drive them. An investigation identified high fungal counts in some problem buses (> 70,000 colony forming units [CFU]/m3; n = 3) compared with control buses (< 220 CFU/m3; n = 4). The predominant species were Penicillium and Cladosporium (1/1). Remedial measures, including washing with a 0.17% solution of sodium hypochlorite and an ozone treatment, were not successful. Because fungal species are heat sensitive, two buses were subjected to a heat treatment of 55 degrees C for 4 hours. In one bus the fungal spores of Cladosporium appeared to be more heat sensitive than the spores of Penicillium. At this point the interior of one bus was completely renewed and another was given a formaldehyde treatment followed by heat treatments. Both strategies reduced fungal counts to 190 from > 107,000 CFU/m3 for the former and to 270 from > 71,000 CFU/m3 for the latter. Only the interior of the most heavily contaminated buses were refurbished prior to the heat treatment, which was done on all problem buses. All buses are still in active service 5 years later. The most frequent health symptoms reported by 88 exposed bus drivers were headache (36%), blocked/runny/itchy nose (26%), nausea (26%), and dry irritated throat (25%). No chronic health effects have been reported after 5 years, although some of these common fungal species are known to be opportunistic pathogens. PMID- 9342834 TI - Assessment of exposure to chemical agents and ergonomic stressors in tanneries in Kanpur, India. AB - In developing countries qualitative assessment of exposure at the workplace may be an essential tool in evaluating hazardous working conditions. This survey reports on qualitative assessment of exposure to chemicals, dust, and ergonomic stressors among 298 workers in 15 tanneries in Kanpur, India. In general, chemical exposure and dermal exposure were highest among beamhouse workers, less for workers involved in dry finishing activities, and lowest for those performing the wet finishing of hides. Dermal exposure was rated as high to very high during beamhouse activities, reflecting direct contact with wet hides and manual handling of hides in soak tanks. Relevant dust exposure was observed only during dry finishing activities. Most workers experienced severe postural load due to working in trunk flexion and rotation for more than 50% of their daily work time. In addition, manual materials handling with loads over 20 kg frequently occurred. The size of the tannery, in general a reflection of state of technology, showed no systematic influence on exposure profiles. The survey suggested that mechanization of material transfer and application of trolleys reduced the work time with trunk flexion and rotation and implied less manual lifting. The presence of local exhaust ventilation in large tanneries seemed to reduce the chemical exposure. This survey has demonstrated the importance of rapid appraisal techniques for evaluating hazardous conditions at the workplace. In developing countries this approach may facilitate occupational hygiene research and practice. PMID- 9342835 TI - Respiratory disorders, skin complaints, and low-back trouble among tannery workers in Kanpur, India. AB - In a cross-sectional survey health complaints among 418 laborers in 15 Indian tanneries were studied. Low-back pain (61%), asthma (38%), dermatitis (23%), and chronic bronchitis (14%) were the most frequently reported complaints in the 12 months prior to the survey. In general, beamhouse workers reported the highest prevalence but only chronic low-back pain was significantly elevated compared with workers in the finishing departments. When using individual exposure estimates, clear associations were presented among manual lifting over 20 kg and low-back pain (OR = 3.5) and skin exposure and dermatitis (OR = 2.6). Frequent lifting of loads was also associated with self-reported asthma. About 44% of the laborers reported at least one period of sickness absence, and 17% were involved in a serious occupational accident that required a visit to the local physician. Logistic regression analysis showed that sickness absence occurred more often in small tanneries (OR = 2.7) and also was significantly associated with low-back pain (OR = 3.3) and occupational accidents (OR = 2.2). This epidemiologic survey on health complaints in tannery workers is among the few in occupational populations in low-income countries. For many reasons these populations are easily overlooked. The results of this descriptive study indicate that there is a clear need for epidemiologic surveys in these countries to obtain information on working conditions and associated health problems. PMID- 9342836 TI - Monitoring and analysis of occupational exposure to chain saw exhausts. AB - The extent of inhalation exposure to loggers from two-stroke chain saws was measured and evaluated under various conditions. Carbon monoxide, measured by personal air monitoring and determination of carboxyhemoglobin levels of the loggers, was used as an indicator of exhaust exposure. Video recordings were made to analyze the influence of varying working conditions and the individual handling of the chain saw on the amount of pollutants inhaled. The American Conference of Governmental Industrial Hygienists biological exposure index (BEI) for carboxyhemoglobin (3.5%) was exceeded during logging in heavy timber stands. When workers were paid on a piecework basis, carboxyhemoglobinemia increased to its maximum level in the first 2-3 hours of the shift and then declined. After 8 hours carboxyhemoglobin levels were 20-30% lower compared with the maximum. Increased exhaust inhalation with short-term exposures to carbon monoxide up to 400 ppm was observed in the following conditions: (1) felling operations, (2) other operations performed in a leaning of squatting position, (3) limbing in thick tops of coniferous trees, (4) working at low wind velocity, and (5) working in thick forest stands. Maximum allowable blood concentrations for carboxyhemoglobin are exceeded in chain saw operators in logging operations. Blood sampling at the end of the workday is not always suitable for determining the highest carboxyhemoglobin levels in loggers during the shift. The exposure of chain saw operators to exhaust increases under certain conditions. PMID- 9342837 TI - Effect of personal hygiene on blood lead levels of workers at a lead processing facility. AB - The relationship between personal hygiene and blood lead levels was tested at a lead processing facility. During the workers' semiannual respirator fit test, when they were confident their hands were clean, the amount of lead on their right hands was measured. Samples were obtained by cleaning one entire hand with a wiping towel treated with a proprietary mixture of alcohol, surfactants, and ethylenediaminetetraacetic acid. Wipe samples were analyzed for total lead and then compared with the worker's blood lead level. Each worker's personal habits at rest were also observed. Workers with more than 1 year's experience had a significantly positive correlation between lead on the hand tested and their blood level. The study strongly suggests that lead on the skin ultimately enters the bloodstream. The route of entry was not investigated. Personal habits of the workers with high blood lead levels were observed to include actions that would quickly contaminate their hands shortly after washing. PMID- 9342838 TI - Synthesis of the Ca(2+)-dependent cell adhesion molecule DdCAD-1 is regulated by multiple factors during Dictyostelium development. AB - In Dictyostelium discoideum, the cadA gene encodes the cell adhesion molecule DdCAD-1, a protein of M(r) 24,000, which mediates Ca(2+)-dependent cell-cell adhesion during development. We have examined the effects of cAMP, cell-cell contact, and growth conditions on cadA expression. cadA has a unique pattern of expression, which appears to be a combination of the expression patterns of early genes and aggregation-stage genes. Expression of the cadA gene in bacterially grown cells is activated at the beginning of the developmental cycle, followed by a period of rapid DdCAD-1 accumulation. The mRNA level reaches its maximum at 9 h of development and then declines to the basal level at approximately 18 h, while the protein level remains constant after reaching its maximum at 12 h. Pulse chase experiments have demonstrated that DdCAD-1 has a significantly longer half life than the average cellular protein. Transcription of the cadA gene is stimulated by exogenous cAMP pulses, leading to a 3- to 5-fold increase in the transcription rate. In the fgdA mutant, which lacks a functional G alpha 2, cAMP fails to enhance cadA expression, suggesting that cAMP stimulates cadA transcription via a G protein-dependent pathway. However, inhibition of cell-cell contact has no effect on the synthesis of DdCAD-1. Growth conditions also have a major influence on cadA expression. Axenically grown cells produce a high level of cadA transcripts during vegetative growth. The mRNA level shows a steady decrease during development and is reduced to the basal level by 12 h. In contrast, the level of DdCAD-1 remains relatively high throughout development, suggesting that axenic growth affects the accumulation of cadA mRNA but not the stability of the protein. These results indicate that multiple mechanisms are involved to maintain a high level of DdCAD-1 during development. PMID- 9342839 TI - Simultaneous demonstration of lens regeneration from dorsal iris and tumour production from ventral iris in the same newt eye after carcinogen administration. AB - It is well known that urodeles have the most powerful regenerative capacities among vertebrates, but there is little realisation that they are extremely resistant to spontaneous or chemically induced tumours. Regeneration and carcinogenesis have been considered to be two sides of the same mechanism. Since antagonism between regeneration and carcinogenesis was expected in previous studies, the present study was intended to clarify this relationship in greater detail by changing the amounts of carcinogen stepwise. When 1 microliter nickel subsulfide solution was administered in various amounts (1 microgram/microliter approximately 40 micrograms/microliter) into lentectomized newt eyes, the delay of initiation in lens regeneration for 6 months and an increased inhibition rate of lens regeneration at 6 months were observed in proportion to the increase in carcinogen dosage. The tumour production rate increased in accordance with the increase in the amounts of carcinogen. The conspicuous result obtained in the present study was that lens regeneration from dorsal iris and tumour induction from ventral iris occurred simultaneously in the same eye after administration of moderate amounts (10 micrograms/microliter) of carcinogen. These data clearly indicated that the regenerating dorsal iris is persistently resistant to carcinogen, whereas the ventral iris, which cannot regenerate lens, is susceptible to tumour induction. This strongly suggests that the lens regeneration system in the newt has special advantages for research on the relationship between regeneration and carcinogenesis. PMID- 9342840 TI - Identification and localization of a neurally expressed member of the plakoglobin/armadillo multigene family. AB - The plakoglobin/armadillo multigene family comprises many proteins widely differing in sizes and functions which have in common a variable number of tandemly repeated arm sequences of about 42 amino acids (aa). In a search for proteins with sequence homology to the desmosomal-plaque-associated arm-repeat containing protein, plakophilin 1, we have identified a novel plakoglobin/armadillo protein. This new member of the multigene family is predominantly, if not exclusively, expressed in neural and neuroendocrine tissues, hence the name neural plakophilin-related arm-repeat protein (NPRAP). The murine cDNA codes for a protein of 1247 aa, with a predicted molecular weight of 135 kDa and a pI of 7.57. The orthologous human protein differs only in a few aa, indicative of the evolutionary stability of NPRAP. In human and murine cDNAs, we have found different transcripts of the NPRAP gene, suggesting that in each species the protein exists in at least two isoforms. The NPRA protein contains three different regions: a 528-aa amino-terminal "head" domain, including a potential coiled-coil-forming alpha-helix segment, a central domain with 10 imperfect arm-repeat units, and a 212-aa carboxy-terminal "tail" domain. By aa sequence, NPRAP is highly homologous to three proteins: p120cas, p0071 and ARVCP, which represent a distinct subgroup within the plakoglobin/armadillo family. By in situ hybridization and immunofluorescence microscopy using NPRAP-specific antibodies, we have demonstrated NPRAP and its mRNA in the perikarya of various kinds of CNS neurons in embryonic and adult mice, but minimal amounts have also been detected by immunoblot analysis in some other tissues containing neural or neuroendocrine elements. We have not seen significant enrichment of NPRAP at cell junctions or in nuclei. Possible NPRAP functions are discussed and the correlation of NPRAP synthesis with neuronal differentiation processes is emphasized. PMID- 9342841 TI - Cytokeratin expression in human arteries pertinent to intimal thickening formation in the ductus arteriosus. AB - Expression of epithelial cytokeratins type 8, 18 and 19 can be used to study smooth muscle cell differentiation during development. We studied the differentiation of smooth muscle cells in the ductus arteriosus before and during intimal thickening and compared the changes occurring in this vessel with the adjoining elastic ascending and descending aorta and the pulmonary trunk. The ductus arteriosus, a vessel connecting the pulmonary trunk and the aorta during fetal life, constricts shorty after birth and eventually closes. Effective closure occurs only in the case of well developed intimal thickening. Cytokeratin expression during fetal development was greatest in the media of the ascending aorta and pulmonary artery, while in the ductus and descending aorta cytokeratin staining was slight. These results suggest that ductus smooth muscle cells and the smooth muscle cells of the descending aorta show a more advanced differentiation as compared to the ascending aorta and pulmonary artery. At neonatal stages cytokeratin expression in the descending aorta, pulmonary artery and the ascending aorta had disappeared as was expected with increased differentiation. In the neonatal ductus arteriosus reexpression of cytokeratins was found in cell clusters in the hyaluronic acid rich environment of the intimal thickening and in the inner media. Reexpression of cytokeratins, especially when organized in clusters, may reflect changes in gene regulation. Therefore the clusters of cytokeratin positive cells in the ductus may be indicative of extensive changes, occurring during closure of this vessel in the neonatal period, in which inner media and intima are especially involved. PMID- 9342842 TI - Growth inhibition of cultured human gastric cancer cells by 9-cis-retinoic acid with induction of cdk inhibitor Waf1/Cip1/Sdi1/p21 protein. AB - The effect of 9-cis-retinoic acid (9-cis-RA) on the growth of eight gastric cancer cell lines was related to their transcription levels of mRNAs for retinoid receptors. Northern blot analysis showed that seven (TMK-1, MKN-1, -28, -45, -74, HSC-39, KATO-III) out of eight gastric cancer cell lines synthesized mRNAs for retinoic acid receptors (RARs) and retinoid X receptor-alpha (RXR-alpha). MKN-7 cells did not transcribe either RARs or RXR-alpha at the mRNA level although they appeared to have no alterations at the gene level. The growth of all of the cell lines except for MKN-7 cells was inhibited by 1 x 10(-6) M 9-cis-RA. Cell cycle distribution analysis revealed that G0-G1 arrest was not induced by exposure to 9 cis-RA in the sensitive TMK-1 and KATO-III cells or the resistant MKN-7 cells. Interestingly, 9-cis-RA temporarily increased the amount of the cyclin dependent kinase (cdk) inhibitor, Waf1/Cip1/Sdi1/p21 protein, and also reduced the amount of cdk-7, epidermal growth factor receptor (EGFR) and cyclin D1 proteins, followed by reduction in phosphorylation of the product of the retinoblastoma tumor suppressor gene (Rb) in the sensitive TMK-1 cells, but not in the resistant MKN-7 cells. These results suggest that 9-cis-RA has a cytostatic effect on gastric cancer cells that synthesize the receptor molecules through cell cycle regulatory machinery. PMID- 9342843 TI - Common defects of different retinoic acid resistant promyelocytic leukemia cells are persistent telomerase activity and nuclear body disorganization. AB - The acute promyelocytic leukemia (APL) t(15;17) rearrangement fuses the promyelocytic leukemia (PML) gene to the retinoic acid receptor-alpha (RAR alpha). There is expression of the chimeric transcript, PML/RAR alpha, in these APL cells. These clinical APL cases respond to the differentiation agent all trans retinoic acid (ATRA) with complete but not durable remissions because ATRA resistance develops. The NB4 APL cell line expresses PML/RAR alpha and responds to the growth inhibitory and differentiation-inducing signals of ATRA. To identify mechanisms responsible for ATRA resistance in APL, ATRA-resistant NB4 cell lines were derived from parental NB4 cells using different strategies. These lines were resistant to the growth inhibition and differentiation effects of ATRA. ATRA-resistant cells were isolated as a de novo resistant line from parental NB4 cells (NB4-R1), following chemical mutagenization and selection in ATRA (NB4-R2), or after chronic selection in ATRA (NB4-R3). Common defects linked to this ATRA resistance were found. When cultured in ATRA, these resistant cells still express PML, RAR alpha, and PML/RAR alpha proteins. Sequence abnormalities were not detected in the RAR alpha DNA binding domains cloned from a representative RA-resistant NB4 line. In ATRA-sensitive but not ATRA-resistant NB4 cells, ATRA down-regulated retinoid X receptor-alpha (RXR alpha) expression, a known marker of ATRA response in parental NB4 cells. Notably, engineered overexpression of RXR alpha in ATRA-sensitive NB4 cells did not block ATRA mediated growth suppression. ATRA treatment of these resistant NB4 lines did not signal a decline in telomerase activity or reorganization of PML-associated nuclear bodies, but both events occurred in ATRA-sensitive NB4 cells. These ATRA resistant NB4 lines are not fully differentiation-defective, since monocytic maturation was induced following treatment with phorbol 12-myristate 13-acetate (PMA) and 1,25 dihydroxy vitamin D3 (vitamin D3). Notably, induced monocytic differentiation of these distinct ATRA-resistant APL lines markedly repressed telomerase activity. Thus, this study suggests that persistent telomerase activity and nuclear body disorganization are linked to ATRA resistance in APL. PMID- 9342844 TI - Environmental stress and the expression of genetic variation. AB - We have started to test the effects of environmental extremes on the expression of genetic variation for traits likely to be under selection in natural populations. We have shown that field heritability may be high for stress response traits in contrast to morphological traits, which tend to show lower levels of heritable variation in nature compared with the laboratory. Selection for increased stress resistance can lead to a number of other evolutionary changes, and these may underlie trade-offs between favourable and stressful environments. Temperature extremes can have a marked influence on the heritability of life history traits. Heritabilities for fecundity can be high when parental flies are reared at low temperatures and under field conditions. The expression of genetic variation for development time is somewhat more complex when temperature extremes are considered. Populations at species margins may be ideal for studying the effects of environmental stress on evolution. PMID- 9342845 TI - Worldwide latitudinal clines for the alcohol dehydrogenase polymorphism in Drosophila melanogaster: what is the unit of selection? AB - Geographical clines may reflect the action of natural selection on genetic polymorphisms. In Drosophila melanogaster several latitudinal clines occur for many characters like allozymes, inversions and quantitative traits. The identical nature of these clines on the various continents, both on the Northern and Southern Hemispheres strongly suggests adaptation to specific stress factors. The alcohol dehydrogenase (Adh) polymorphism shows high frequencies of the S allele in tropical regions and declines with latitude. The reasons for this cline are difficult to determine because of the entanglement with other polymorphisms varying with latitude. In this paper the tentative connections with other polymorphisms like alpha-Gpdh, In(2L)t, body size and development time are reviewed with respect to the possible environmental stress factors involved. It is concluded, also from recent experiments, that the (2L)t inversion plays a dominant role in resistance to high temperature and is partly responsible for the Adh cline. Further research is aimed at the specific selective forces acting on Adh, focussing on the physiological and life history aspects. PMID- 9342846 TI - Stress and metabolic regulation in Drosophila. AB - Environmental changes that result in stress (defined here as decreased absolute viability and/or fecundity) result in extrinsic changes in metabolism that are to some extent compensated by altered gene expression. The fact that different genotypes may respond differently to environmental stress may be of key importance to the maintenance of genetic variation in metabolic traits. Here we quantify a set of metabolic characters in genetically defined lines of Drosophila melanogaster subjected to four stresses (3% acetic acid, 3% ethanol, starvation and thermal stress) in order to assess the magnitude of environmental effects and genotype x environment interactions. Genetic correlations were quantified, and many exhibit significant heterogeneity across environments. Pleiotropically related traits may exhibit the phenomenon of apparent selection, whose effects may be particularly strong in stressful environments. This transient apparent selection may have a large consequence on the maintenance of genetic variation. PMID- 9342847 TI - Phenotypic and evolutionary adaptation of a model bacterial system to stressful thermal environments. AB - We studied both phenotypic and evolutionary adaptation to various thermal environments using the bacterium Escherichia coli as an experimental model system. We determined that 42 degrees C was stressful to a bacterial clone adapted to 37 degrees C, based on reductions in both absolute and competitive fitness, as well as induction of a heat stress response. This clone was also used to found replicated populations that were propagated for thousands of generations under several different thermal regimes, including 42 degrees C. Evolutionary adaptation of the populations to 42 degrees C resulted in an increase in both absolute and relative fitness at that temperature, measured respectively as an increase in the number of descendants (and their biovolume) and in competitive ability relative to the ancestral clone. The replicated experimental lineages achieved their evolutionary improvement by several distinct pathways, which produced differential preadaptation to a non-stressful nutrient environment. Adaptation to this stressful temperature entailed neither a change in the ancestral thermal niche nor any pronounced trade-offs in fitness within the thermal niche, contrary to a priori predictions. This study system was several important advantages for evaluating hypotheses concerning the effects of stress on phenotypic and evolutionary adaptation, including the ability to obtain lineages that have evolved in controlled and defined environments, to make direct measurements of fitness and to quantify the degree of stress imposed by different environments. PMID- 9342848 TI - Ecological and evolutionary physiology of heat shock proteins and the stress response in Drosophila: complementary insights from genetic engineering and natural variation. AB - Classical adaptational and genetic engineering approaches offer complementary insights to understanding biological variation: the former elucidates the origins, magnitude and ecological context of natural variation, while the latter establishes which genes can underlie natural variation. Studies of the stress or heat shock response in Drosophila illustrate this point. At the cellular level, heat shock proteins (Hsps) function as molecular chaperones, minimizing aggregation of peptides in non-native conformations. To understand the adaptive significance of Hsps, we have characterized thermal stress that Drosophila experience in nature, which can be substantial. We used these findings to design ecologically relevant experiments with engineered Drosophila strains generated by unequal site-specific homologous recombination; these strains differ in hsp70 copy number but share sites of transgene integration. hsp70 copy number markedly affects Hsp70 levels in intact Drosophila, and strains with extra hsp70 copies exhibit corresponding differences in inducible thermotolerance and reactivation of a key enzyme after thermal stress. Elevated Hsp70 levels, however, are not without penalty; these levels retard growth and increase mortality. Transgenic variation in hsp70 copy number has counterparts in nature: isofemale lines from nature vary significantly in Hsp70 expression, and this variation is also correlated with both inducible thermotolerance and mortality in the absence of stress. PMID- 9342849 TI - High-temperature stress and the evolution of thermal resistance in Drosophila. AB - The evolution of thermal resistance and acclimation is reviewed at the population level using populations and isofemale lines of Drosophila buzzatii and D. melanogaster originating from different climatic regions. In general, ample genetic variation for thermal resistance was found within and among populations. A rough correlation between the climate of origin and thermal resistance was apparent. Acclimation at a non-lethal temperature led to a significant increase in survival after heat shock, and recurrent acclimation events generally increased survival even further. Acclimation effects lasted over several days, but this effect decreased gradually with time since acclimation. Protein studies showed that the concentration of Hsp70 in adult flies is greatly increased by acclimation and thereafter gradually decreases with time. For populations with relatively high survival at one life stage, survival often was low at other life stages. Furthermore, selection on different life stages showed that a selection response in one life stage did not necessarily result in a correlated response in another. These observations indicate that different mechanisms or genes at least in part are responsible for or are expressed at different developmental stages. Selection for increased resistance was successful despite low heritabilities for the trait. Survival and fertility were compared between acclimated and non acclimated flies, and a cost of expressing the "heat shock response" was identified in that increased survival after acclimation was accompanied by reduced fertility. The relative costs increased under nutritional stress. Metabolic rate was genetically variable but did not correlate with temperature resistance. The more resistant lines, however, often had shorter developmental time. Inbreeding reduced thermal stress tolerance of adult flies, but it did not reduce tolerance of embryos that possibly are exposed to strong natural selection for thermal stress resistance. In general, inbreeding may reduce stress resistance, and thus multiple stressful events may account for increased inbreeding depression in harsh environments. PMID- 9342850 TI - Genetic and environmental stress, and the persistence of populations. AB - Many populations of endangered species have to cope both with stressful and deteriorating environmental conditions (mostly the primary cause of the endangerment) and with an increase in homozygosity due to genetic drift and/or inbreeding in small isolated populations. The latter will result in genetic stress often accompanied by a decrease in fitness (inbreeding depression). We have studied the consequences of genetic stress, under optimal as well as stressful environmental conditions, for the fitness and persistence of small populations using Drosophila melanogaster as a model system. The results show that, already under optimal environmental conditions, an increase in homozygosity or inbreeding both impairs fitness and increases the extinction risk of populations significantly. Under environmental stress, however, these effects become greatly enhanced. More important, the results show that the impact of environmental stress becomes significantly greater for higher inbreeding levels. This explicitly demonstrates that genetic and environmental stress are not independent but can act synergistically. This apparent interaction may have important consequences for the conservation of endangered species. PMID- 9342851 TI - Adaptation and extinction in changing environments. AB - The extinction risk of a population is determined by its demographic properties, the environmental conditions to which it is exposed, and its genetic potential to cope with and adapt to its environment. All these factors may have stochastic as well as directional components. The present chapter reviews several types of models concerned with the vulnerability of small populations to demographic stochasticity and to random and directional changes of the environment. In particular, the influence of mutation and genetic variability on the persistence time of a population is explored, critical rates for environmental change are estimated beyond which extinction on time scales of tens to a few thousand generations is virtually certain, and the extinction risks caused by the above mentioned factors are compared. PMID- 9342852 TI - Environmental stress and evolution: a theoretical study. AB - A haploid population subject to recurrent deleterious mutations and two environments that provide two different profiles of selection coefficients over loci are modeled. The population is supposed to inhabit one "home" environment where it evolves the corresponding genetic constitution. One generation of the population is then exposed to the second, "foreign" environment. The decline in the mean population fitness is considered as a measure of stress in the population caused by the foreign environment. I define the relative strength of stress as 1 minus the ratio of the mean fitnesses in the foreign and home environments, and give the corresponding analytical expression. The stress strength is composed of three different contributors: the environmental component of stress, which is determined by purely external, non-genetic factors of the foreign environment (its carrying capacity), and two genetic components. The latter consists of the environment x genetic component, caused by the direct influence of the foreign environment on selection coefficients, and of the evolutionary component that is due to the adaptation of the population to the home environment. Among others, it is shown that even if the home and foreign environments were equivalent, so that both the environmental and environmental x genetic components of stress were absent, stress would still occur in the foreign environment due to the evolutionary component. The model also predicts that stressful foreign environments cause an increase in the genotypic variance of fitness. Some other features of population variability in stressful environments are discussed. A general conclusion that can be drawn from this model is that a certain environment may be claimed as stressful only if considered with respect to both a given population and the environment in which the population has evolved. PMID- 9342853 TI - Genetic variability and adaptation to stress. AB - Besides an immediate cellular adaptation to stress, organisms can resist such challenges through changes in their genetic material. These changes can be due to mutation or acquisition of pre-evolved functions via horizontal transfer. In this chapter we will review evidence from bacterial genetics that suggests that the frequency of such events can increase in response to stress by activating mutagenic response (e.g. the SOS response) and by inhibiting antimutagenic activities (e.g. mismatch repair system, MRS). Natural selection, by favoring adaptations, can also select for the mechanism(s) that has/have generated the adaptive changes by hitchhiking. These mutator mechanisms can sometimes respond very specifically, though blindly, to the challenge of the environment. Such stress-induced increases in mutation rates enhance genetic polymorphism, which is the structural component of the barrier to genetic exchange. Since SOS and MRS are the enzymatic controls of this barrier, the modulation of these systems can lead to a burst of speciation. PMID- 9342854 TI - Stress-resistance genotypes, metabolic efficiency and interpreting evolutionary change. AB - Assuming stress levels to which free-living populations are normally exposed, an association between rapid development time, a long life, success in mating and size of sexual ornaments can be predicted. Fitness at one stage of the life cycle should therefore correlate with fitness at other stages under this environmental model. Assuming that stress targets energy carriers, high-energy efficiency underlain by stress-resistance genotypes that are likely to be heterozygous is the basis of this prediction. Stress-resistance genotypes therefore have a role in promoting the energy efficiency required for organisms to accommodate a stressed world. Selection for energy efficiency to utilize heterogenous resources implies that the process of speciation should normally occur rapidly and be rarely observed. It follows that the ecological species concept is primary to other species concepts. The intensity of selection for stress resistance goes from an extreme in the highly disturbed and stressful environments of living fossils to relatively stable abiotic habitats, where specialist diversifications and adaptive radiations are likely. Between these extremes, a punctuated pattern of evolutionary change may occur in perturbed environments during a transient phase of increased resources. In abiotically benign tropical habitats where energy constraints are low, specialization of resource utilization by learning appears possible. PMID- 9342855 TI - Invariant association of ecdysis with increases in cyclic 3',5'-guanosine monophosphate immunoreactivity in a small network of peptidergic neurons in the hornworm, Manduca sexta. AB - At the end of each molt insects shed their old cuticle by performing the stereotyped behavior of ecdysis. In the moth, Manduca sexta, this behavior is triggered by the neuropeptide eclosion hormone (EH). Insights into the mechanism of action of EH have come from the identification of a small network of peptidergic neurons that shows increased cyclic 3',5'-guanosine monophosphate (cGMP) immunoreactivity at ecdysis in insects from many different orders. Here we present further evidence that strengthens the association between ecdysis and the occurrence of this cGMP response in Manduca. We found that the cGMP increases occurred at every ecdysis, although some of the neurons that showed a response at larval ecdysis did not participate at pupal and adult ecdysis. Both ecdysis and the cGMP increases only required an intact connection with the brain for the first 30 min after EH injection. Interestingly, ecdysis in debrained animals only occurred if the cGMP response had been initiated, suggesting that the onset of this response marks the time at which the central nervous system is first able to drive ecdysis. Finally, we found that the appearance of sensitivity to EH for triggering the cGMP response coincided with the time at which EH first triggers ecdysis. PMID- 9342857 TI - Maximal biomass of Arabidopsis thaliana using a simple, low-maintenance hydroponic method and favorable environmental conditions. PMID- 9342856 TI - GABAergic disinhibition changes the recovery cycle of bat inferior collicular neurons. AB - This study examines the contribution of GABAergic inhibition to the discharge pattern and recovery properties of 110 bat inferior collicular neurons by means of bicuculline application to their recording sites. When stimulated with single pulses, 74 (67%) neurons discharged one or two impulses (phasic responders), 19 (17%) discharged three to ten impulses (phasic bursters) and 17 (16%) discharged impulses throughout the entire stimulus duration (tonic responders). Bicuculline application changed phasic responders into phasic bursters or tonic responders, increased the number of impulses by 10-2000% and shortened the response latency of most neurons. When stimulated with pairs of sound pulses, the recovery cycles of these neurons can be described as: (1) long inhibition (n = 49, 45%); (2) short inhibition (n = 41, 37%); and (3) fast recovery (n = 20, 18%) based upon the 50% recovery time that was either longer than 20 ms, between 10 and 20 ms or shorter than 10 ms. Bicuculline application shortened the 50% recovery time of most neurons by 11-2350% allowing them to respond to pairs of sound pulses at very short interpulse intervals. These data demonstrate that GABAergic inhibition contributes significantly to auditory temporal processing. PMID- 9342860 TI - Epigenetic transcriptional silencing and 5-azacytidine-mediated reactivation of a complex transgene in rice. AB - Despite a growing number of reports indicating non-Mendelian inheritance of transgene expression in monocots, no detailed description of the structure and stability of the transgene exists for transformants generated by direct DNA transfer techniques, making the cause for these observations difficult to determine. In this paper we describe the complex organization of Btt cryIIIA and bar transgenes in rice (Oryza sativa L.) that displayed aberrant segregation in R1 progeny. Silencing rather than rearrangement of the bar gene was implicated because the herbicide-sensitive R1 plants had a DNA hybridization profile identical to that of the resistant R0 parent and R1 siblings. Genomic DNA analysis revealed substantial methylation of the Ubi1/bar sequences in silenced plants and, to a lesser degree, in herbicide-resistant plants, suggesting that the transgene locus was potentiated for silencing. Nuclease protection and nuclear run-on assays confirmed that silencing was due to transcriptional inactivation. Treatment of R2 progeny of silenced plants with 5-azacytidine resulted in demethylation of the Ubi1 promoter and reactivation of bar gene expression, demonstrating a functional relationship for methylation in gene silencing. These findings indicate that methylation-based silencing may be frequent in cereals transformed by direct DNA protocols that insert multiple, often rearranged sequences. PMID- 9342861 TI - Promoter analysis and expression of a phospholipase D gene from castor bean. AB - The expression of a castor bean (Ricinus communis L.) phospholipase D (PLD; EC 3.1.4.4) gene has been studied by examining its promoter activity in transgenic tobacco (Nicotiana tabacum) carrying a PLD promoter-glucuronidase transgene and by monitoring the levels of PLD mRNA in castor bean. Sequence and the 5' truncation analyses revealed that the 5' flanking region from nucleotide -1200 to -730 is required for the regulation and basal function of the PLD promoter. The PLD promoter in vegetative tissues is highly active in the rapidly growing regions such as the shoot apex and the secondary meristem producing axillary buds and vascular tissues of young leaves and stems. The PLD promoter activity in floral tissues was high in stigma, ovary, and pollen grains, but low in petals, sepals, the epidermis of anthers, styles, and filaments. The PLD promoter activity was enhanced by abscisic acid. Northern-blot analysis of PLD in castor bean showed that the PLD mRNA levels were high in young and metabolically more active tissues such as expanding leaves, hypocotyl hooks, developing seeds, and young seedlings, and they decreased in mature tissues such as fully expanded leaves and developed seeds. These patterns of expression suggest a role of PLD in rapid cell growth, proliferation, and reproduction. PMID- 9342863 TI - The massugu1 mutation of Arabidopsis identified with failure of auxin-induced growth curvature of hypocotyl confers auxin insensitivity to hypocotyl and leaf. AB - Unilateral application of indole-3-acetic acid (IAA) in a lanolin base to hypocotyls of partially etiolated seedlings of wild-type Arabidopsis thaliana induced growth curvature in a dose-dependent manner. The effects of IAA in concentrations from 1 to 1000 microM were studied, with maximum IAA-induced curvature at 100 microM. Three IAA-insensitive mutants were isolated and are all in the same locus, massugu1 (msg1). They did not undergo hypocotyl growth curvature at any of the IAA concentrations tested. msg1 is recessive and is located on chromosome 5. msg 1 hypocotyl growth is resistant to 2,4 dichlorophenoxyacetic acid (2,4-D), but the roots are as sensitive to 2,4-D as the wild type. Growth of the hypocotyl was inhibited to essentially the same extent as the wild type by 6-benzylaminopurine, abscisic acid, and 1 aminocyclopropane-1-carboxylate, an ethylene precursor. The msg1 leaves were also resistant to 2,4-D-induced chlorosis. The gravitropic response of the msg1 hypocotyl takes much more time to initiate and achieve the wild-type degree of curvature, whereas the msg1 roots responded normally to gravity. The mature plants and the etiolated seedlings of msg1 were generally wild type in appearance, except that their rosette leaves were either epinastic or hyponastic. msg1 is the first auxin-insensitive mutant in which it effects are mostly restricted to the hypocotyl and leaf, and msg1 also appears to be auxin specific. PMID- 9342865 TI - Gene expression in the pulp of ripening bananas. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products and cDNA cloning of 25 different ripening-related mRNAs. AB - mRNA was extracted from the pulp and peel of preclimacteric (d 0) bananas (Musa AAA group, cv Grand Nain) and those exposed to ethylene gas for 24 h and stored in air alone for a further 1 (d 2) and 4 d (d 5). Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products from the pulp and peel of these fruits revealed significant up-regulation of numerous transcripts during ripening. The majority of the changes were initiated by d 2, with the level of these messages increasing during the remainder of the ripening period. Pulp tissue from d 2 was used for the construction of a cDNA library. This library was differentially screened for ripening-related clones using cDNA from d-0 and d-2 pulp by a novel microtiter plate method. In the primary screen 250 up- and down-regulated clones were isolated. Of these, 59 differentially expressed clones were obtained from the secondary screen. All of these cDNAs were partially sequenced and grouped into families after database searches. Twenty five nonredundant groups of pulp clones were identified. These encoded enzymes were involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation, and several other key metabolic events. We describe the analysis of these clones and their possible involvement in ripening. PMID- 9342864 TI - Regulation of sesquiterpene cyclase gene expression. Characterization of an elicitor- and pathogen-inducible promoter. AB - The promoter for a tobacco (Nicotiana tabacum) sesquiterpene cyclase gene, a key regulatory step in sesquiterpene phytoalexin biosynthesis, has been analyzed. The EAS4 promoter was fused to the beta-glucuronidase (GUS) reporter gene, and the temporal and spatial expression patterns of GUS activity were examined in stably transformed plants and in transient expression assays using electroporated protoplasts of tobacco. No GUS activity was observed in any tissues under normal growth conditions. A low level of GUS activity was detected in wounded leaf, root, and stem tissues, whereas a much higher level was observed when these tissues were challenged with elicitors or microbial pathogens. The GUS expression pattern directed by the EAS4 promoter was identical to the induction patterns observed for the endogenous sesquiterpene cyclase genes. Neither exogenous salicylic acid nor methyl jasmonate induced GUS expression; and H2O2 induced GUS expression to only a limited extent. Although the EAS4 promoter contains cis sequences resembling previously identified transcriptional control motifs, other cis-sequences important for quantitative and qualitative gene expression were identified by deletion and gain-of-function analyses. The EAS4 promoter differs from previously described pathogen-/elicitor-inducible promoters because it only supports inducible gene expression and directs unique spatial expression patterns. PMID- 9342866 TI - Differential gene expression in ripening banana fruit. AB - During banana (Musa acuminata L.) fruit ripening ethylene production triggers a developmental cascade that is accompanied by a massive conversion of starch to sugars, an associated burst of respiratory activity, and an increase in protein synthesis. Differential screening of cDNA libraries representing banana pulp at ripening stages 1 and 3 has led to the isolation of 11 nonredundant groups of differentially expressed mRNAs. Identification of these transcripts by partial sequence analysis indicates that two of the mRNAs encode proteins involved in carbohydrate metabolism, whereas others encode proteins thought to be associated with pathogenesis, senescence, or stress responses in plants. Their relative abundance in the pulp and tissue-specific distribution in greenhouse-grown banana plants were determined by northern-blot analyses. The relative abundance of transcripts encoding starch synthase, granule-bound starch synthase, chitinase, lectin, and a type-2 metallothionein decreased in pulp during ripening. Transcripts encoding endochitinase, beta-1,3-glucanase, a thaumatin-like protein, ascorbate peroxidase, metallothionein, and a putative senescence-related protein increased early in ripening. The elucidation of the molecular events associated with banana ripening will facilitate a better understanding and control of these processes, and will allow us to attain our long-term goal of producing candidate oral vaccines in transgenic banana plants. PMID- 9342867 TI - Identification of promoter elements involved in the cytosolic Ca(2+)-mediated photoregulation of maize cab-m1 expression. AB - Changes in cytoplasmic Ca2+ levels are involved in the regulation of several plant genes. However, to our knowledge, no regions of genes or specific cis elements have been shown to be involved in the regulation of plant gene expression by cytosolic Ca2+ signaling. The maize (Zea mays) gene cab-m1, which encodes a light-harvesting chlorophyll a/b-binding apoprotein, is positively photoregulated in mesophyll cells (MC) but not in bundle-sheath cells (BSC). This gene is highly preferentially expressed in maize MC versus BSC. In situ transient expression assays have revealed that exposure of tissues to ethyleneglycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA), which chelates Ca2+, blocks the photostimulation of cab-m1 full promoter (-1026 to + 14) activity in MC of leaf segments of dark-grown maize seedlings. EGTA has no effect on expression in BSC. These results suggest that light-induced elevation of the cytosolic Ca2+ concentration in MC is required for the enhancement of cab-m1 expression in MC. Deletion of the sequence from -1026 to -360 completely abolished Ca2+ responsiveness of cab-m1 expression in MC. On the other hand, a 54 bp fragment in the 5' flanking region (-953 to -899 relative to the translation start site) conferred Ca2+ responsiveness on a -359 core promoter: reporter gene, suggesting that Ca2+ signaling is mediated via specific sequences in this short fragment. Furthermore, possible involvement of Ca(2+)-calmodulin in the signal transduction chain for regulating cab-m1 expression was suggested by the results of inhibitor experiments. PMID- 9342868 TI - Sequence analysis of the cloned glossy8 gene of maize suggests that it may code for a beta-ketoacyl reductase required for the biosynthesis of cuticular waxes. AB - The gl8 locus of maize (Zea mays L.) was previously defined by a mutation that reduces the amount and alters the composition of seedling cuticular waxes. Sixty independently derived gl8 mutant alleles were isolated from stocks that carried the Mutator transposon system. A DNA fragment that contains a Mu8 transposon and that co-segregates with one of these alleles, gl8-Mu3142, was identified and cloned. DNA flanking the Mu8 transposon was shown via allelic cross-referencing experiments to represent the gl8 locus. The gl8 probe revealed a 1.4-kb transcript present in wild-type seedling leaves and, in lesser amounts, in other organs and at other developmental stages. The amino acid sequence deduced from an apparently full-length gl8 cDNA exhibits highly significant sequence similarity to a group of enzymes from plants, eubacteria, and mammals that catalyzes the reduction of ketones. This finding suggests that the GL8 protein probably functions as a reductase during fatty acid elongation in the cuticular wax biosynthetic pathway. PMID- 9342869 TI - Anion-channel blockers interfere with auxin responses in dark-grown Arabidopsis hypocotyls. AB - Anion channels are thought to participate in signal transduction and turgor regulation in higher plant cells. The regulation of hypocotyl cell elongation is a situation in which these channels could play important roles because it involves ionic fluxes that are implicated in turgor control and orchestrated by various signals. We have used a pharmacological approach to reveal the contribution of anion channels in the regulation of the development of hypocotyls by auxins. Auxins induce an inhibition of elongation, a disintegration of the cortical cell layers, and the formation of adventitious roots on Arabidopsis thaliana hypocotyls grown in the dark. Anion-channel blockers such as anthracene 9-carboxylic acid, 4,4'-diisothiocyanatostilbene-2-2'-disulfonic acid, 4 acetamido-4'-isothiocyanato-stilbene-2-2'-disulfonic acid, and R(+) methylindazone; indanyloxyacteic acid-94, which produce little or no stimulation of hypocotyl elongation by themselves, are able to counteract the inhibition and the disintegration induced by auxins with various efficiencies. This interference appears to be specific for auxins and does not occur when hypocotyl elongation is inhibited by other growth regulators such as ethylene or cytokinins. The putative involvement of anion channels in auxin signal transduction is discussed. PMID- 9342870 TI - Two homologous low-temperature-inducible genes from Arabidopsis encode highly hydrophobic proteins. AB - We have characterized two related cDNAs (RCI2A and RCI2B) corresponding to genes from Arabidopsis thaliana, the expression of which is transiently induced by low, nonfreezing temperatures. RCI2A and RCI2B encode small (54 amino acids), highly hydrophobic proteins that bear two potential transmembrane domains. They show similarity to proteins encoded by genes from barley (Hordeum vulgare L.) and wheatgrass (Lophophyrum elongatum) that are regulated by different stress conditions. Their high level of sequence homology (78%) and their genomic location in a single restriction fragment suggest that both genes originated as a result of a tandem duplication. However, their regulatory sequences have diverged enough to confer on them different expression patterns. Like most of the cold inducible plant genes characterized, the expression of RCI2A and RCI2B is also promoted by abscisic acid (ABA) and dehydration but is not a general response to stress conditions, since it is not induced by salt stress or by anaerobiosis. Furthermore, low temperatures are able to induce RCI2A and RCI2B expression in ABA-deficient and -insensitive genetic backgrounds, indicating that both ABA dependent and -independent pathways regulate the low-temperature responsiveness of these two genes. PMID- 9342873 TI - Characterization of regions within the N-terminal 6-kilodalton domain of phytochrome A that modulate its biological activity. AB - Phytochrome A (phyA) is a red/far-red (FR) light photoreceptor responsible for initiating numerous light-mediated plant growth and developmental responses, especially in FR light-enriched environments. We previously showed that the first 70 amino acids of the polypeptide contain at least two regions with potentially opposite functions (E.T. Jordan, J.R. Cherry, J.M. Walker, R.D. Vierstra [1996] Plant J 9: 243-257). One region is required for activity and correct apoprotein/chromophore interactions, whereas the second appears to regulate phytochrome activity. We have further resolved these functional regions by analysis of N-terminal deletion and alanine-scanning mutants of oat (Avena sativa) phyA in transgenic tobacco (Nicotiana tabacum). The results indicate that the region involved in chromophore/apoprotein interactions contains two separate segments (residues 25-33 and 50-62) also required for biological activity. The region that regulates phyA activity requires only five adjacent serines (Sers) (residues 8-12). Removal or alteration of these Sers generates a photoreceptor that increases the sensitivity of transgenic seedlings to red and FR light more than intact phyA. Taken together, these data identify three distinct regions in the N-terminal domain necessary for photoreceptor activity, and further define the Ser-rich region as an important site for phyA regulation. PMID- 9342876 TI - A dynamin-like protein, ADL1, is present in membranes as a high-molecular-mass complex in Arabidopsis thaliana. AB - Dynamin, a GTP-binding protein, is involved in endocytosis in animal cells. We found that a dynamin-like protein, ADL1, is present in multiple forms in Arabidopsis leaf tissue. Subcellular fractionation experiments, together with gel filtration and nondenaturing-gel electrophoresis revealed that most of ADL1 is present as a high-molecular-mass complex of 400 to 600 kD in the membrane or pellet fraction, whereas ADL1 is present in the soluble fraction as a monomer. The subcellular distribution of ADL1 is affected by various agents such as Ca2+, cyclosporin A, GTP, and ATP. Ca2+ increases the amount of ADL1 present in the membrane fraction, whereas cyclosporin A inhibits the membrane association. Furthermore, Ca2+ and GTP change the migration pattern of ADL1 in nondenaturing polyacrylamide gels, indicating that these chemicals influence either the complex formation and/or the conformation of the ADL1 complex. Our results demonstrate that ADL1 has characteristics that are similar to Dynamin I, which is found in animal cells. Therefore, it is possible that ADL1 is also involved in biological processes that require vesicle formation. PMID- 9342877 TI - Two rhamnogalacturonide tetrasaccharides isolated from semi-retted flax fibers are signaling molecules in Rubus fruticosus L. cells. AB - Water extraction of semi-retted flax (Linum usitatissimum L.) fiber bundles yielded a mixture of pectic oligosaccharides and two acidic rhamnogalacturonide tetrasaccharides that were separated by size-exclusion chromatography. One- and two-dimensional nuclear magnetic resonance studies and fast atom bombardment-mass spectrometry experiments indicated that the two tetrasaccharides have a common primary structure, i.e. alpha-D-delta GalpA(1-->2)-alpha-L- Rhap(1-->4)-alpha-D GalpA-(1-->2)-L-alpha,beta-Rhap, with a rhamnopyranose as terminal reducing end, and a 4-deoxy-beta-L-threo-hex-4-enopyranosiduronic acid at the nonreducing end. However, the two tetrasaccharides differ by an acetyl group located at the O-3 position of the internal galacturonic acid residue. These two tetrasaccharides induce the activation of D-glycohydrolases of Rubus fructicosus L. cells or protoplasts within minutes. PMID- 9342878 TI - Jasmonic acid-dependent and -independent signaling pathways control wound-induced gene activation in Arabidopsis thaliana. AB - Plant response to mechanical injury includes gene activation both at the wound site and systemically in nondamaged tissues. The model developed for the wound induced activation of the proteinase inhibitor II (Pin2) gene in potato (Solanum tuberosum) and tomato (Lycopersicon esculentum) establishes the involvement of the plant hormones abscisic acid and jasmonic acid (JA) as key components of the wound signal transduction pathway. To assess in Arabidopsis thaliana the role of these plant hormones in regulating wound-induced gene expression, we isolated wound- and JA-inducible genes by the differential mRNA display technique. Their patterns of expression upon mechanical wounding and hormonal treatments revealed differences in the spatial distribution of the transcripts and in the responsiveness of the analyzed genes to abscisic acid and JA. A correlation can be established between sensitivity to JA and the accumulation of the transcripts in systemic tissues upon wounding. A comparative study of the wound response in wild-type and JA-insensitive coi1 mutant plants indicated that in A. thaliana wound signals are transmitted via at least two different pathways. One of them does not involve JA as a mediator and is preferentially responsible for gene activation in the vicinity of the wound site, whereas the other requires JA perception and activates gene expression throughout the aerial part of the plant. PMID- 9342879 TI - DNA mismatch repair in plants. An Arabidopsis thaliana gene that predicts a protein belonging to the MSH2 subfamily of eukaryotic MutS homologs. AB - Sets of degenerate oligomers corresponding to highly conserved domains of MutS homolog (MSH) mismatch-repair proteins primed polymerase chain reaction amplification of two Arabidopsis thaliana DNA fragments that are homologous to eukaryotic MSH-like genes. Phylogenetic analysis places one complete gene, designated atMSH2, in the evolutionarily distinct MSH2 subfamily. PMID- 9342880 TI - A water-soluble chlorophyll protein in cauliflower may be identical to BnD22, a drought-induced, 22-kilodalton protein in rapeseed. AB - A water-soluble chlorophyll protein (WSCP) in cauliflower (Brassica oleracea L.) was purified and its N-terminal sequence was determined. Forty-six of 48 residues of the sequence completely matched those of the drought-induced 22-kD protein (BnD22) in rapeseed (Brassica napus L.). All 40 sequenced residues of WSCP from rapeseed were perfectly matched to those of BnD22. Thus, WSCP may be identical to BnD22. The abundance of WSCP was increased in detached cauliflower leaves. PMID- 9342881 TI - The influence of moisture on microbial transport, survival and 2,4-D biodegradation with a genetically marked Burkholderia cepacia in unsaturated soil columns. AB - The influence of moisture on the survival, movement and degradation activity of a 2,4-D degrading bacterium, Burkholderia cepacia strain BRI6001L, genetically engineered to contain bioluminescent and lactose utilization genes, was studied in unsaturated soil columns. The distance traveled by BRI6001L was dependent on the clay content of the soil, higher clay contents being responsible for higher filtration coefficients. Long term survival, in excess of one year, was attributed to strain BRI6001L's ability to survive dry conditions. Changes in the 2,4-D biodegradation rate showed a better correlation with the BRI6001L population density than with the total viable bacterial population. At moisture levels between field capacity and 40% moisture (-33 kPa to -100 kPa) 2,4-D degradation was attributed mainly to BRI6001L. At moisture levels between 6 and 15%, 2,4-D disappearance was attributed to the indigenous microbial population, with no degradation occurring at moisture levels below 6%. Returning the moisture to above 40% led to an increase of 4 orders of magnitude in the BRI6001L population density and to a 10-fold increase in the 2,4-D degradation rate. The ability to monitor a specific microbial population using reporter genes has demonstrated the importance of controlling moisture levels for maximizing biodegradation rates in unsaturated soil environments. PMID- 9342882 TI - Genetic engineering of bacteria and their potential for Hg2+ bioremediation. AB - Ion exchange or biosorptive processes for metal removal generally lack specificity in metal binding and are sensitive to ambient conditions, e.g. pH, ionic strength and the presence of metal chelators. In this study, cells of a genetically engineered Escherichia coli strain, JM109, which expresses metallothionein and a Hg2+ transport system after induction were evaluated for their selectivity for Hg2+ accumulation in the presence of sodium, magnesium, or cadmium ions and their sensitivity to pH or the presence of metal chelators during Hg2+ bioaccumulation. The genetically engineered E. coli cells in suspension accumulated Hg2+ effectively at low concentrations (0-20 microM) over a broad range of pH (3 to 11). The presence of 400 mM sodium chloride, 200 mM magnesium chloride, or 100 microM cadmium ions did not have a significant effect on the bioaccumulation of 5 microM Hg2+, indicating that this process is not sensitive to high ionic strength and is highly selective against sodium, magnesium, or cadmium ions. Metal chelators usually interfere with ion exchange or biosorptive processes. However, two common metal chelators, EDTA and citrate, had no significant effect on Hg2+ bioaccumulation by the genetically engineered strain. These results suggest that this E. coli strain could be used for selective removal of Hg2+ from waste water or from contaminated solutions which are resistant to common treatments. A second potential application would be to remove Hg2+ from Hg(2+)-contaminated soil, sediment, or particulates by washing them with a Hg2+ chelator and regenerating the chelator by passing the solution through a reactor containing the strain. PMID- 9342883 TI - Use of a luminescent bacterial biosensor for biomonitoring and characterization of arsenic toxicity of chromated copper arsenate (CCA). AB - An arsenic oxyanion-inducible Escherichia coli chromosomal operon (arsRBC) has been previously identified. Construction of a luciferase transcriptional gene fusion (arsB::luxAB) showed that ars operon expression, plus concomitant cell luminescence, was inducible in a concentration-dependent manner by arsenic salts. The present study was conducted to evaluate the potential of the arsB::luxAB transcriptional gene fusion for use as a biosensor in monitoring the toxicity of arsenic compounds. Cultures from this gene fusion strain were exposed to increasing concentrations of the wood preservative chromated copper arsenate (CCA), as well as its constituents, sodium arsenate and chromated copper solution (CC). Analysis of luciferase activity revealed that the arsB::luxAB gene fusion was expressed in response to CCA and sodium arsenate, but not to the CC solution. The detection limit of arsenic was found to be 0.01 microgram As/ml (10 parts per billion, 10 ppb) and therefore well within the range of environmental concerns. A greater induction of luminescence by arsenate was observed when cells were limited for phosphate, as phosphate can act as a competitive inhibitor of arsenate ions. Our results suggest that the E. coli arsB::luxAB fusion strain has a promising future as a specific and sensitive biosensor for monitoring bioavailable levels and toxicity of arsenic near sites where CCA-treated wood has been used. PMID- 9342884 TI - Detection of heavy metal ion resistance genes in gram-positive and gram-negative bacteria isolated from a lead-contaminated site. AB - Resistance to a range of heavy metal ions was determined for lead-resistant and other bacteria which had been isolated from a battery-manufacturing site contaminated with high concentration of lead. Several Gram-positive (belonging to the genera Arthrobacter and Corynebacterium) and Gram-negative (Alcaligenes species) isolates were resistant to lead, mercury, cadmium, cobalt, zinc and copper, although the levels of resistance to the different metal ions were specific for each isolate. Polymerase chain reaction, DNA-DNA hybridization and DNA sequencing were used to explore the nature of genetic systems responsible for the metal resistance in eight of the isolates. Specific DNA sequences could be amplified from the genomic DNA of all the isolates using primers for sections of the mer (mercury resistance determinant on the transposon Tn501) and pco (copper resistance determinant on the plasmid pRJ1004) genetic systems. Positive hybridizations with mer and pco probes indicated that the amplified segments were highly homologous to these genes. Some of the PCR products were cloned and partially sequenced, and the regions sequenced were highly homologous to the appropriate regions of the mer and pco determinants. These results demonstrate the wide distribution of mercury and copper resistance genes in both Gram positive and Gram-negative isolates obtained from this lead-contaminated soil. In contrast, the czc (cobalt, zinc and cadmium resistance) and chr (chromate resistance) genes could not be amplified from DNAs of some isolates, indicating the limited contribution, if any, of these genetic systems to the metal ion resistance of these isolates. PMID- 9342885 TI - Introduction and PCR detection of Desulfomonile tiedjei in soil slurry microcosms. AB - The aim of this work was to test the feasibility of introducing an anaerobic microbial reductive dechlorination activity into non sterile soil slurry microcosms by inoculation with the pure anaerobic bacterial strain Desulfomonile tiedjei, which is capable of dechlorinating 3-chlorobenzoate to benzoate. To show that the bacterium was established in the microcosms we followed the expression of the reductive dechlorination activity and a molecular probe based on PCR amplification of the 16S rDNA gene was developed. However, the success of PCR amplification of the 16S rDNA gene depends on the DNA extraction and purification methodologies applied, as shown through the use of several protocols. In this study we report a DNA extraction and purification method which generates sufficient and very clean DNA suitable for PCR amplification of the D. tiedjei 16S rDNA gene. The threshold of detection was about 5.10(3) bacteria per gram of soil slurry. Introduction of D. tiedjei in soil slurry microcosms proved successful since 3-chlorobenzoate dechlorination activity was established with this bacterium in microcosms normally devoid of this dechlorination capacity. Indeed, the addition of D. tiedjei to microcosms supplemented with acetate plus formate as cosubstrate, at their respective concentrations of 5 and 6 mM, led to a total biotransformation of 2.5 mM of 3-chlorobenzoate within 12 days. After complete 3-chlorobenzoate dechlorination, the 16S rDNA gene of this bacterium was specifically detected only in the inoculated microcosms as shown by PCR amplification followed by restriction mapping confirmation. PMID- 9342886 TI - Detection of STEC and epidemiological investigations in surrounding of a HUS patient. AB - After occurrence of a case of HUS infection in a 2-year-old infant from a dairy farmer's family living near Oldenburg, investigations were performed in the infant's surrounding in order to elucidate the route of infection. Since hospitalization took place at a late stage, it was not possible to isolate EHEC from the patient's stool samples. However, E. coli O157 antibody determinations in serum were positive. Since STEC of serogroup O157 were found in faeces from the 34 dairy cows of the farm, stool samples were taken from 6 members of the child's family and examined. Non-O157 STEC could be isolated from the stools of 2 family members. Determination of other virulence factors and other characteristics such as serotype, biotype and phage type showed identity of the agent for 3 isolates (2 from animals, 1 from humans). By means of pulsed-field gel electrophoresis of the restricted DNA of the isolates and by means of RAPD PCR it was not possible to establish any differences in the band patterns. It can be assumed, therefore, that the organisms had been transmitted from animals to humans. PMID- 9342887 TI - EEG-based communication: evaluation of alternative signal prediction methods. AB - Individuals can learn to control the amplitude of EEG activity in specific frequency bands over sensorimotor cortex and use it to move a cursor to a target on a computer screen. For one-dimensional (i.e., vertical) cursor movement, a linear equation translates the EEG activity into cursor movement. To translate an individual's EEG control into cursor control as effectively as possible, the intercept in this equation, which determines whether upward or downward movement occurs, should be set so that top and bottom targets are equally accessible. The present study compares alternative methods for using an individual's previous performance to select the intercept for subsequent trials. In offline analyses, five different intercept selection methods were applied to EEG data collected while trained subjects were moving the cursor to targets at the top or bottom edge of the screen. In the first two methods-moving average, and weighted sum-a single intercept was selected for the entire 1-2 sec period of each trial. In the other three methods-blocked moving average, blocked weighted sum, and blocked recursive sum (a variation of the weighted sum)-an intercept was selected for each 200-ms segment of the trial. The results from these methods were compared in regard to their balance between upward and downward movements and their consistency of performance across trials. For all subjects combined, the five methods performed similarly. However, performance across subjects was more consistent for the moving average, blocked moving average, and blocked recursive sum methods than for the weighted sum and blocked weighted sum methods. Due to its consistent performance and its computational simplicity, the moving average method, using the five most recent pairs of top and bottom trials, appears to be the method of choice. PMID- 9342889 TI - Current status of respiratory syncytial virus (RSV) and parainfluenza virus type 3 (PIV3) vaccine development: memorandum from a joint WHO/NIAID meeting. AB - A Joint WHO/National Institute of Allergy and Infectious Diseases (NIAID) meeting on the current status of respiratory syncytial virus (RSV) and parainfluenza virus type 3 (PIV3) vaccine development was held in Bethesda, MD, from 30 September to 1 October 1996. The meeting summarized the worldwide impact of RSV and PIV3; presented the current status of development of RSV and PIV3 vaccines; and examined the applications of recombinant DNA technology to the development and characterization of vaccines and to the understanding of viral pathogenesis. PMID- 9342888 TI - The WHO Global Programme for Vaccines and Immunization Vaccine Trial Registry. AB - In 1995, the WHO Global Programme for Vaccines and Immunization established a Vaccine Trial Registry. As of September 1996, this registry included 50 WHO supported vaccine trials, of which 25 (50%) were completed studies. The vaccines most frequently tested have been against measles (9 trials), poliovirus (8 trials), cholera (8 trials), enterotoxigenic Escherichia coli (4 trials), and pneumococcus (4 trials). Nearly 80% of these trials have been conducted in developing countries, with the largest number being in Africa. Among the 25 completed trials, outcomes measured were immune response (24 trials), adverse reactions (13 trials), morbidity (4 trials), and mortality (1 trial). WHO's contributions to these studies include direct funding, assistance with study design, site visits, data analysis, vaccine procurement, and vaccine potency testing. PMID- 9342890 TI - How important are airfreight rates and vaccine packaging in cost-saving efforts for the Expanded Programme on Immunization? AB - Vaccines constitute the single most important cost factor in the Expanded Programme on Immunization (EPI) in Mozambique and in view of future new disease control initiatives, the proportional expenditure on vaccines will only increase. Airfreight may contribute up to at least 25% of the total cost of delivered vaccine. Air transport of vaccine provided by UNICEF was arranged by the vaccine supplier. As a result of a lack of control mechanisms, airfreight rates were unnecessarily high and showed considerable variation. By negotiating rates directly with the airlines, the EPI management team in Mozambique succeeded in reducing them from an average of about US$ 12 per kg to US$ 4 per kg, equivalent to an annual saving of US$ 100,000. Vaccine vials are typically packaged in one of the following types of boxes: small colourful boxes containing 5-10 vials or bigger more functional boxes containing 50-100 vials. The packaging of vials in smaller boxes can double airfreight costs compared with bigger boxes. The EPI management team for Mozambique recommends that UNICEF should take over from suppliers the arrangements for shipping vaccine and negotiate airfreight rates centrally; further, WHO should tighten current vaccine-packaging standards for net packaging volume per dose, so that packaging in uneconomical small boxes can be eliminated. PMID- 9342891 TI - The Ife South Breastfeeding Project: training community health extension workers to promote and manage breastfeeding in rural communities. AB - Reported are the results of a project to promote exclusive breastfeeding in rural communities through the training of community health extension workers in rural Nigeria. A workshop for the trainers was organized for health workers in the study area; subsequently, these trainers ran district-level training workshops. In the study area perinatal facilities, early initiation of breastfeeding has increased compared with those in the control area (P < 0.001). Also, the trained health workers had significantly better knowledge about breastfeeding than their untrained colleagues in both the study (P < 0.001) and control areas (P < 0.001), and more often recommended timely initiation and exclusive breastfeeding than the controls (P < 0.001). A multivariate analysis showed that the training programme and the study area were the only significant variables that were predictors of breastfeeding knowledge (P < 0.001). Appropriate education of health extension workers can therefore contribute significantly to the promotion of breastfeeding in rural communities. PMID- 9342892 TI - The development of a MUAC-for-height reference, including a comparison to other nutritional status screening indicators. AB - Mid-upper-arm circumference (MUAC) based on a single cut-off value for all the children less than 5 years of age has been used for many years as an alternative nutritional status index for children during famines or refugee crises, and as an additional screening tool in nonemergencies. However, it has recently been questioned whether MUAC is age- and sex-independent. After reviewing the scientific evidence underlying the use and interpretation of MUAC, a WHO Expert Committee recommended a new MUAC-for-age reference for under-5-year-olds. In some settings, however, it is difficult to assess a child's age and in such circumstances MUAC-for-height may be a good alternative. The height-based QUAC stick is a simple means of adjusting MUAC cut-offs according to height, and the MUAC-for-height reference and the construction and use of the QUAC stick are described in this article. Described also is the use of the receiver operating characteristic (ROC) curve method to evaluate the performance of MUAC, MUAC-for age, and MUAC-for-height in screening malnourished children. PMID- 9342893 TI - In vivo evaluation of sixteen plant extracts on mice inoculated with Trypanosoma brucei gambiense. AB - After examination of the drugs used by traditional practitioners in Cote d'lvoire, nine formulas prescribed in the treatment of African human trypanosomiasis (AHT) were selected for investigation. These formulas made use of 40 plants, 16 of which were studied because of their properties, as described in the literature, and their frequent use by practitioners. The plant extracts were administered, after maceration or decoction, either orally or intraperitoneally to Swiss mice that had previously been inoculated with Trypanosoma brucei gambiense (Tbg), strain MHOM/Cl/81/Dal 083. The parasitaemia in each mouse was followed for three consecutive days and compared with that in control mice, which had been given either a saline solution (SS: negative control) or well-known drugs (melarsoprol, difluoromethylornithine, and pentamidine: positive control). Our investigations led to the following conclusions. (a) None of the plant extracts revealed trypanocidal or trypanostatic activity relative to SS controls (P > 0.05). In fact, the mice that received the extracts died on the third day after inoculation, with 0% survival and an average parasitaemia of 10.8 +/- 2 x 10(7) trypanosomes/ml. (b) The treated positive controls, relative to SS, showed 100% survival and no parasitaemia (P < 0.05). Melarsoprol appeared to be active when given orally at a dose of 3.6 mg/kg body weight twice a day for 3 days. This method of testing the sensitivity of trypanosomes to plant extracts is easy and inexpensive, and could be applied to other areas of research on tropical diseases. PMID- 9342894 TI - Serotypes of group A streptococci isolated from healthy schoolchildren in the United Arab Emirates. AB - Group A streptococci (GAS) are the most frequent cause of pharyngitis in children and are a common cause of emergency room or paediatric clinic visits worldwide. This study determined the representative M and T types of GAS, and their distribution, among schoolchildren in the United Arab Emirates. Throat swabs were taken and cultured for GAS isolates during the winter of 1994-95 from 1000 children aged 5-7 years attending nine schools. Of the isolates obtained, 100 were serotyped using standard techniques. Nearly all these isolates (91%) were T typable, falling into 15 T types; the commonest being type 1 (n = 17), type 6 (n = 15), type 11 (n = 10), type 2 (n = 8), type 12 (n = 8), and type 28 (n = 8). A total of 76% of the isolates were typable for M protein, falling into 14 M types, with type 1 (n = 17), type 6 (n = 15), type 2 (n = 8), type 22 (n = 5), type 28 (n = 7), and type 75 (n = 5) predominating. Serotype clusters were found in certain classes or schools, although the number of isolates examined was too small to allow definitive epidemiological conclusions to be drawn. The ease of serotyping these isolates suggests that GAS strains in the United Arab Emirates are similar, but not necessarily related, to those commonly found in the USA and Europe, and that these may be the most prevalent strains worldwide. The relative prevalence of M type 1 is significant, as this GAS serotype is associated with serious diseases such as rheumatic heart disease, a recognized problem in the United Arab Emirates, and toxic shock syndrome, which has not yet been reported from this area. Knowledge of the prevalence of GAS serotypes, and further research on the epidemiology of streptococcal disease, will be useful should streptococcal vaccines become available. PMID- 9342895 TI - Diagnosis of disseminated mycobacterial infection: testing a simple and inexpensive method for use in developing countries. AB - With the development of the acquired immunodeficiency syndrome (AIDS) epidemic, the isolation of mycobacteria from blood has become a common problem for clinical laboratories. In this study two methods were used for the recovery of mycobacteria from blood specimens obtained from AIDS patients: (1) direct inoculation of a biphasic medium, and (2) a non-commercial lysis-centrifugation method. A total of 3 consecutive blood samples were taken at 15-minute intervals from each of 50 AIDS patients with clinical suspicion of disseminated mycobacterial disease. Mycobacterium growth was noted in 70/138 blood specimens from 30 (60%) patients. These cultures yielded Mycobacterium tuberculosis in 19 (63%) and Mycobacterium avium complex organisms in 11 (37%) patients. Cultures using the lysis-centrifugation method were positive in 54% of the patients while cultures using biphasic medium were positive in 44% (P > 0.05). The positivity for M. avium complex was higher with lysis-centrifugation (91%) than with biphasic medium (45.4%) (P < 0.05). However, the positivities for M. tuberculosis with the lysis-centrifugation method (89.5%) and direct inoculation in biphasic medium (100%) were similar (P > 0.05). The use of a non-commercial lysis centrifugation technique is inexpensive, reliable, and can be an alternative method for the diagnosis of mycobacteraemia in developing countries. PMID- 9342897 TI - The utility of DALYs for public health policy and research: a reply. AB - The WHO Advisory Committee on Health Research (ACHR), through its DALY Review Group, has recently criticized the use of disability-adjusted life years (DALYs). To suggest that the use of DALYs should be discouraged as an aid in health resource allocation may, however, be premature, since it enhances informed debate on the social values that influence resource allocation, identifies health problems that may be neglected, and points to the strengths and weaknesses of existing health information systems. PMID- 9342896 TI - Strategies for minimizing nosocomial measles transmission. AB - As a result of the highly contagious nature of measles before the onset of rash, nosocomial transmission will remain a threat until the disease is eradicated. However, a number of strategies can minimize its nosocomial spread. It is therefore vital to maximize awareness among health care staff that an individual with measles can enter a health facility at any time and that a continual risk of the nosocomial transmission of measles exists. The present review makes two groups of recommendations: those which are generally applicable to all countries, and certain additional recommendations which may be suitable only for industrialized countries. PMID- 9342898 TI - The critique of DALYs: a counter-reply. AB - The DALY Review Group of the WHO Advisory Committee on Health Research (ACHR) believes that, unless they are constructed purely as an intellectual exercise, indicators should have a function--ultimately to guide decision-making about resource allocation. Disability-adjusted life years (DALYs) obscure too much and in its present stage of development the DALY approach does not solve the problem of prioritization and of resource allocation and may yet turn out to have been a side-track. PMID- 9342899 TI - CD antigens 1996: updated nomenclature for clusters of differentiation on human cells. IUIS/WHO Subcommittee on CD Nomenclature. PMID- 9342900 TI - Primary surgery of skeletal dysgnathias. AB - Three case studies from the patient population of the University Hospital of Aachen (RWTH) are used to describe indications for primary surgical intervention in skeletal dysgnathia. Such preconditions may apply in the case of mandibulo alveolar protrusion, anomalies where there is little or no option to fixing orthodontic appliances (enamel hypoplasia or shortened crowns), severe transversal discrepancies and skeletal dysgnathia with pronounced malpositioning in the alveolar process and the teeth. The advantages are improved compliance (through the patient experiencing success at the outset of treatment) and limitation of postoperative orthodontic treatment to occlusive fine adjustments of the occlusion, resulting in an appreciable reduction in both the degree and duration of tooth movement and tissue damage. PMID- 9342902 TI - Outcome prediction of the anterior open bite. Comparison of computer and clinician analysis of cephalograms. AB - Anterior open bite (AOB) may be expected to close spontaneously in approximately 50% of Caucasian patients. Computer-derived discriminant analysis of a single cephalometric radiograph has been shown to predict closure or non-closure in 88% of patients at the pre-puberal stage, in 74% at the puberal stage, and in 94% at the post-puberal stage. In this study, the predictive capacity of the computer analysis was tested against predictions made by groups of clinicians in Belfast and Toronto. The computer analysis was carried out on the first cephalometric radiographs of a new sample of 34 open bite cases collected serially and recorded over a minimum of 2 years. Thus, the spontaneous outcome was known to the authors. The first radiographs were shown to 20 clinicians in Belfast and 22 in Toronto who were asked to predict the spontaneous outcome. The computerised discriminant analysis made correct predictions in 85% and the clinicians in 64% of the sample. There were no significant differences between the predictions of clinicians in Belfast and Toronto, but computer prediction was more accurate than all grades of clinician. The predictions of qualified orthodontists were generally more accurate than prequalified orthodontists which were more accurate than those of undergraduate dental students but the differences did not rise to the level of statistical significance. For patients at the puberal stage the predictive capacity of qualified orthodontists was less than orthodontists in training. Computer prediction of the spontaneous outcome in open bite improves clinical diagnosis. PMID- 9342901 TI - Electronic measurements of relative tongue-palate contact time. Development and testing for orthodontic functional analysis. AB - The importance of the tongue to the form of the jaws and dental arches has long been accepted. Clear-cut differences in arch width and arch height are observed between mouth and nasal breathing. Course measurements, e.g. duration of tongue contact with the gum, are not feasible with traditional measuring methods. The palatal measuring appliance presented here together with the purpose-developed storage and evaluation equipment permits for the first time continuous 24-hour measurement of tongue contact with the palate. The clinical observation is confirmed by the presented results. Nasal obstruction is associated with lower tongue-palate contact times. In our probands, these times fell by an average of 72% after forced mouth breathing. Since complex movements within the mouth cavity cannot be directly observed, functional analysis relating to the tongue position was previously impossible, at least over a longer period. The measuring device presented here is suitable for analyzing in more detail the diagnostically difficult complex of tongue movements and breathing habits. It might therefore conceivably be used to assess myofunctional disturbances and therapeutic methods. PMID- 9342903 TI - Unilateral cleft lip and palate. Relationship between morphology of the dentition and functional parameters of the tongue. AB - The relationship between orthodontic and logopedic findings was evaluated with statistical contingency analysis. The investigation was focused on selected dentomorphologic parameters and oral function/malfunction of 100 patients between 3 and 7 years of age with unilateral cleft lip and palate. For further classification into contingency tables "normal" and "abnormal" attributes were defined. Interdependencies between observed findings and their probable frequencies were calculated and represented in diagrams by means of statistical parameters such as the contingency coefficient. Scatterplots of the calculated coefficients showed a characteristic pattern corresponding to clinical experience. While overjet and transversal occlusion are strongly associated with oral functions, overbite tends not to be associated with oral functions. The contingency coefficient calculated from sagittal and transversal orthodontically relevant conditions of occlusion and incorrect tongue position as well as incorrect swallowing tends to be stronger than that of "abnormal" morphologic parameters and an incorrect tongue position during speech. The longitudinal study of lip position and the 3 investigated morphologic parameters revealed strongly associated contingency coefficients. PMID- 9342904 TI - Replantation of an inverted lower second premolar germ. AB - During the initial X-ray control of a 10-year-old female orthodontic patient, a late development and an inverted position of the lower left second premolar was seen. 4.3 years later the tooth with its follicle was replanted in the upright position, preserving the second deciduous molar; at that time the root development was at an early stage. At a follow-up control 4 years after the surgical procedure the replanted tooth had erupted and the sensibility response was normal. Its root, however, although completed, could not reach the length and especially the width (in the apical half) of the contralateral premolar. This relative reduction in width seems to follow a temporary thickening of that part of the root formed during the first period after replantation. The anamnestic data suggest that physiologic exfoliation of the deciduous molar took place. PMID- 9342905 TI - Indication and frequency of X-rays in connection with orthodontic treatment. Statement by the Deutsche Gesellschaft fur Kieferothopadie. PMID- 9342906 TI - Report on 9th Interdisciplinary Symposium of the Workshop Cleft Lip and Palate. Morphology and functions of the nose. PMID- 9342907 TI - Stroke rehabilitation in the 90s. PMID- 9342909 TI - Carotid endarterectomy: the "gold standard" for prevention of stroke from severe carotid stenosis. PMID- 9342908 TI - Anticoagulation for stroke prevention in a Medicare population with atrial fibrillation. PMID- 9342910 TI - Advances in stroke therapy: introduction to cerebral angioplasty and cerebral thrombolysis. PMID- 9342912 TI - Evolution in emergency stroke care. National and Florida stroke initiatives. PMID- 9342911 TI - Stroke prevention: a medical obligation. PMID- 9342913 TI - Emergency medical services and the acute stroke: changing the paradigm. PMID- 9342915 TI - Reduction of cancer mortality and incidence by selenium supplementation. AB - PATIENTS AND METHOD: In order to test the hypothesis that a dietary supplement of selenium (Se) may reduce cancer risk, 1312 patients with histories of basa/squamous cell carcinomas of the skin were assigned in random, double-blind fashion to daily oral supplements of either Se-enriched yeast (200 micrograms Se/day), or a low-Se yeast placebo. Patients were recruited in 1983 to 1990 and were followed with regular dermatologic examinations through, 1993 for a total of 8269 person-years of observation. Skin cancer diagnoses were confirmed histologically and plasma Se concentration was determined at 6 to 12 months intervals. All deaths and patient-reported illnesses were confirmed and documented by consultation with the patient medical care providers. RESULTS: Results showed that Se-supplementation did not significantly affect the incidences of recurrent basal/squamous cell carcinomas of the skin. However, Se treatment was associated with reductions in total cancer mortality and in the incidences of lung, colorectal, prostate and total cancers. These effects were consistent over time and between study clinics. CONCLUSION: The results strongly suggest benefits of Se-supplementation for this cohort of patients and support the hypothesis that supplemental Se can reduce risks to at least some types of cancer. PMID- 9342917 TI - TB or not TB? Increasing door-to-door response to screening. AB - Tuberculosis (TB) has made a disconcerting come-back in the United States in recent years. In 1994, Texas ranked third nationally in total number of TB cases and fifth in annual TB case rates. This is of great concern to the Texas Department of Health (TDH) and has led to the development of TB Innovative Demonstration Projects under the Tuberculosis Elimination Division of the TDH. One such project involves identifying high-risk communities by utilizing a computer-based geographic information system and then sending field-workers door to-door offering free skin testing. Because this project was so successful in identifying positive skin test reactors, numerous requests have been made to duplicate its methods. One area of improvement is to increase individual and family participation. The purpose of this article is to present a survey of the literature on nonresponse in door-to-door soliciting, analyze the project's methods of soliciting and nonresponse, and propose ways nonresponse can be decreased in future projects of this nature. PMID- 9342916 TI - Environmental health hazards: the impact on a southern community. AB - The impact of environment on health is a growing area of concern for nurses. The purpose of this study is to describe and characterize residents living near an abandoned hazardous waste site, assess their past and present exposure risk, and describe health concerns and psychosocial effects. Data were obtained by a door to-door outreach effort. Networking was also used to identify residents with health concerns. Extensive field notes were taken to record the responses of residents. A preliminary windshield survey was conducted to approximate the number of residents living in the geographic area, assess the residential community, analyze drainage pathways away from the site, and observe outdoor activity patterns. Community concerns from the residential population were collected and analyzed for recurrent themes. Significant off-site activity patterns included use of drainage ditches as play areas for children. Foraging on site activities included picking berries, crawfishing, geophagy (clay eating), and rabbit hunting. Common health problems described by residents included skin rashes, respiratory-related complaints, reproductive difficulties, and headaches. The majority of residents were concerned about the incidence of cancer in the area. Identifying the concerns of residents and indicators of health problems helps nurses begin to understand the relationship between the environment and health. PMID- 9342918 TI - Rural dwellers' cancer fears and perceptions of cancer treatment. AB - The purpose of this study was to explore differences in cancer fears and perceptions of cancer treatment among four rural groups: men with cancer, men caregivers, women with cancer, and women caregivers (N = 590). The four groups differed in their cancer fears. About half or less feared pain, nausea, body disfigurement, and sexual problems from cancer. Over two-thirds were worried about finances and decreases in quality of life. More worried about separation from loved ones than worried about death. The four groups differed only slightly in perceptions of treatment. The majority thought chemotherapy, surgery, and radiation were important treatments. A larger percentage saw nutritional interventions and biologicals as unimportant in cancer prevention or treatment. Nurses need to address fears and perceptions of cancer treatment with persons experiencing cancer and their families. In addition, these persons and their families need support for decisions regarding cancer treatment. PMID- 9342919 TI - Policy advocacy for public health practitioners: workshops on policy change. AB - This article describes two workshops on health-promoting policy change, developed and delivered to more than 1,000 public and community health practitioners across one Canadian province. The workshops were designed to increase knowledge about the process of policy change and provide opportunities for skill development. Three basic instructional methods--minilectures, individual pen-and-paper exercises, and small-group activities--were used. Workshops were delivered using a host organization arrangement, meaning organizations willing to host workshops could do so if they provided the venue and invited members of other organizations in the community. Initial evaluations indicate that practitioners find the workshops useful and that many have integrated the information into their day-to day work. To the author's knowledge, no other workshops of this nature have been delivered in this systematic way. PMID- 9342920 TI - Tried, true, and new: public health nursing in a county substance abuse treatment system. AB - The Milwaukee Target Cities (MTC) project was the only site within 19 federally funded Target Cities programs to feature a public health nursing model as its sole means of providing comprehensive health-related services to indigent substance abuse clients. We first describe MTC's implementation process, focusing on the public health nursing component, and then present a program evaluation section with selected findings from the ongoing qualitative evaluation. Initially, misunderstandings about the nurses' community-based, family-centered strategy of assuring access to health care through cross-system service linkage dogged the nurses' efforts to explain their roles and mission to federal funders, project management, coworkers, and treatment providers. In the end, after federal funding ended, public health nursing left an enduring legacy of partnerships in the county substance abuse treatment system: education about public health nursing, networking, referral processes, and resources to meet the complex health related needs of indigent substance abusers. Despite the project's many changes, the nurses (a) became specialists in substance abuse, gaining expertise and recognition in a new community, particularly with isolated subpopulations; (b) assured substance abuse clients and their families access to health-related resources through core public health nursing skills; and (c) educated project staff, administrators, providers, and clients about public health nursing. PMID- 9342921 TI - Communication types and sexual protective practices of college women. AB - Sexually transmitted disease, such as the fatal HIV disease, continues to threaten the health of young women. Adolescents have typically been inconsistent users of effective strategies that prevent the spread of disease during sexual activity. Communication practices related to disease-related sexual protection have received little attention as a variable separate from sexual protection. The purpose of this study was to explore the relationship among different types of interpersonal communication of young women and their new sexual partners and their implementation of disease-related sexual protection. A sample of 163 young women who had experienced sexual intercourse completed the Safe Sex Behavior Questionnaire. Interpersonal communication was explored using the general information-seeking (getting to know a partner), sexual self-disclosure, and specific-disease risk-information-seeking scales. These scales were combined; factor analysis revealed three subscales closely related to the original scales. Findings suggest that young women who seek specific information about their new sexual partner's disease risk status are more likely to implement sexual protective practices. These findings provide guidance to nurses who work with adolescent women in advising on aspects of interpersonal relationships that need to be enhanced to protect one's health status. PMID- 9342922 TI - Resources, stigma, and patterns of disclosure in rural women with HIV infection. AB - A growing number of cases of HIV infection are being diagnosed in rural communities especially among women. Although HIV-specific education and care delivery programs have been focused on rural areas in recent years, limited data are available on the impact of such initiatives on the lives of women with HIV infection. The purpose of this study was to examine characteristics of women with HIV disease living in rural communities. The study used a cross-sectional sample of rural women in Georgia. Data analysis indicated that although a majority of the women reported adequate resources, there was a group of women for whom resources for basic needs were not always adequate. Additionally, women with HIV who had not progressed to AIDS had greater difficulty in obtaining a number of resources. Almost half of the women felt stigmatized due to having HIV. Yet, a high percentage of these women had disclosed their HIV status to health care workers, sexual partners, and family. Study results provide insight into the needs of HIV-infected rural women from their perspective. This information can be important to nurses working in public health and community settings as they face the challenge of developing effective health care services for this population. PMID- 9342923 TI - Child-focused single home visiting. AB - Home visiting is a central, long-standing, and yet theoretically underdeveloped public health nursing process. The general aim of this study was to expand and refine a preliminary model of home visiting. A stylized field research investigation was conducted in the area of maternal-child health with one nurse in a visiting nurse association in New England. A specific type of home visiting, identified as child focused, emerged, with phases labeled as surveying and designating; selling and scheduling; approaching the home and the visit; entering the home; gaining permission to ask questions and access the infant--starting with the mother's expressed concerns; making the caregiving judgment--asking questions and weighing and examining the infant; and ending the visit. "Haunting and telling" was an additional phase for certain visits. The nurse conducted child-focused home visiting in three patterns. The single pattern is described in this article. Potential maternal, child, interactive, and environmental consequences were identified, as were factors influencing the process of maternal child home visiting. Social exchange theory emerged as useful in describing how the nurse initiated, maintained, and ended the home visiting process, and in describing attendant client consequences. PMID- 9342924 TI - Immunoreactivity of canine mammary neoplasms with monoclonal antibody CC49. AB - Monoclonal antibody (MAb) CC49 binds to human tumour-associated glycoprotein termed TAG-72. CC49 is a second-generation MAb with higher affinity to TAG-72 than the original MAb B72.3. CC49 was applied to 42 samples from different canine mammary tumours, belonging to seven different histopathological types. Immunoreactivity was detected by the use of an avidin-biotin complex immunoperoxidase method. Most sections from all types of mammary neoplasm reacted with this MAb. Normal tissue did not stain or stained only weakly. The results of this study suggest CC49 has selective immunoreactivity for a variety of canine mammary tumours, which seems superior to that reported with MAb 72.3. These results support the proposal for further study of diagnostic and therapeutic uses of CC49 in the management of canine mammary tumours. PMID- 9342925 TI - Effect of rapeseed feedstuffs with different glucosinolate content and iodine administration on gestating and lactating sow. AB - In two experiments with a total of 60 sows during late pregnancy and at 28 days of lactation, diets containing rapeseed were compared with rapeseed free diets (control). In Experiment 1 dietary content of solvent extracted rapeseed meal was 250 g/kg (10 mmol glucosinolates/kg diet), in Experiment 2 diets containing 100 g/kg rapeseed were tested (2 mmol glucosinolates/kg diet). During late pregnancy all sows received 150 micrograms supplementary iodine/kg diet. In lactation, different subgroups received different rates of iodine administration (Exp. 1:0, 100 or 1000 micrograms/kg diet; Exp. 2: 0, 150 or 300 micrograms/kg diet). Rapeseed feeds had no significant effect on feed intake, body weight of sows and rearing parameters in both experiments. There was a tendency (8%) toward lower litter weight at weaning in Exp. 1. Sow diets without supplementary iodine but containing glucosinolates (via rapeseed meal, rapeseed) caused significant reduction in thyroxine serum concentration of piglets, whereas this hormone did not change in sow serum. Thiocyanate was significantly increased in the serum of mothers. The minor increase of thiocyanate concentration of milk and piglets' serum points to negligible transfer of rapeseed glucosinolate degradation products to offspring. However, the milk iodine concentration was significantly decreased due to glucosinolates, and this seems to be the reason for impaired iodine and thyroid hormone status of piglets from sows given rapeseed feeds. PMID- 9342926 TI - Influence of feeding hay on the alopecia of breeding guinea pigs. AB - Animals with partial alopecia were seen frequently in a breeding colony of guinea pigs. No pathologic aetiology could be determined. An influence of nutrition on the density of the hair coat was considered. Breeding groups were fed one of the commercial guinea pig diets of differing composition, with or without the addition of hay. Observation occurred over a period of months and the quality of the hair coat was evaluated periodically using a semi-quantitative scoring system. More extensive and more frequent fur defects were found is guinea pigs receiving a breeding diet with a high content of crude protein (23%) and a low level of crude fibre (12%), offered hay only restrictively compared with animals receiving a diet with a lower content of crude protein (15.5%) and a higher level of crude fibre (19.5%), offered the same amount of hay. The amount of hay offered is of paramount importance. Animals fed only the diet with the higher level of crude fibre (19.5%), according to the manufacturer's instructions as a complete food, without the addition of hay, showed a less dense hair coat within 4 weeks. In our colony a group of five breeding animals and their young required 200 g of hay daily in order to improve their hair coat quality to normal. Conversely, animals receiving less hay had progressively deteriorating hair coat density. Not only the amount of hay offered, but also the accessibility for all animals plays a role in preventing alopecia in guinea pigs. In larger cages (twice the usual ground surface area) fur defects were seen when the same amount of hay (200 g) was offered in only one central area, rather than spread out evenly throughout the cage. Hair loss was observed to be the result of trichophagia between adult animals kept in the same cage. The need for crude fibre of breeding animals does not appear to be completely met by pellets rich in fibre segments. PMID- 9342927 TI - Inhibition of antigen-induced muscle contractions by inhibitors of thromboxane pathway in rat small intestine. AB - Rats were sensitized against egg albumin and the response of the longitudinal muscle from the proximal small intestine to the antigen was tested. Egg albumin (1-100 micrograms/ml) concentration-dependently induced a contraction of the longitudinal muscle in tissues from sensitized animals but not from nonsensitized animals. The response to the antigen was resistant to neuronal blockers like tetrodotoxin, atropine and hexamethonium. Inhibitors of thromboxane synthesis such as the cyclooxygenase inhibitor, indomethacin, the thromboxane synthase blocker, 1-benzylimidazole, or the combined cyclooxygenase/lipoxygenase/thromboxane synthase inhibitor, sulfasalazine, inhibited the contraction evoked by egg albumin. A similar concentration dependent inhibition of the antigen response was observed with two thromboxane A2 receptor blockers, SK&F 88046 and KW-3635. None of these blockers affected the response to the muscarinic agonist, carbachol, excluding unspecific effects of the drugs on smooth muscle contractility. The effect of antigen was reduced by the mast cell stabilizing agent, quercetin, and by the histamine H1 receptor blocker, mepyramin. These drugs, however, also inhibited the response to carbachol. When contractions were stimulated directly by the stable thromboxane derivative, carbocyclic thromboxane A2, the smooth muscle proved to be more than three orders of magnitude more sensitive to this agonist of the thromboxane pathway compared to histamine. Consequently, thromboxane A2 seems to be one of the main mediators of anaphylactically induced longitudinal muscle contractions in the rat small intestine. PMID- 9342928 TI - The quantitative effect of metoclopramide on abomasal and duodenal myoelectric activity of goats. AB - Electromyographic (EMG) recordings of the abomasal corpus, pyloric antrum and proximal duodenum were made from six goats for 2 h periods before and after administration of 0.5 mg/kg metoclopramide intravenously or intramuscularly. Analog EMG signal was transformed via a computer program to digital data. The percentage change in electrical activity was determined by comparing the electrical activity following administration of IV or IM metoclopramide with the electrical activity of the control periods for the abomasal corpus, pyloric antrum and proximal duodenum. Metoclopramide caused a significant, time-dependent increase in duodenal electrical activity following either route of administration. This increase in duodenal electrical activity coincided with peak plasma levels of metoclopramide until its decline below 100 ng/ml in plasma. There was a significant biphasic increase in electrical activity of the abomasal corpus and pyloric antrum following IM administration of metoclopramide. The first phase lasted approximately 5 min and was followed by a longer period (approximately 20 min) of diminished electrical activity. A second phase of increased electrical activity occurred approximately 40-60 min after initial IM injection of metoclopramide. It is uncertain whether this increase was drug mediated or endogenously-triggered. Similar increases in corpus and antral electrical activity were present following IV metoclopramide administration, though early increases were not statistically significant. Overall, the percentage changes in electrical activity correlated well with predicted peak plasma levels of metoclopramide only in the duodenum. This correlation was limited to approximately 5 min after IV and 15 min after IM metoclopramide administration. PMID- 9342929 TI - Selenite and selenium yeast as feed supplements for dairy cows. AB - The availability of inorganic and organic forms of selenium to dairy cows was studied by giving 25 cows supplementary selenium for 9 months either as sodium selenite or as a selenium-containing yeast product. Group I (eight cows) received 3.0 mg selenium as sodium selenite daily, group II (nine cows) received 3.0 mg selenium as the selenium yeast product, and group III (eight cows) received 0.75 mg selenium as the selenium yeast product. The total selenium contents of the ration were 0.26-0.32 mg/kg feed dry matter for groups I and II, and 0.16-0.18 mg/kg for group III. The supplement of 0.75 mg selenium daily from the yeast product maintained the selenium concentrations of whole blood and milk at the same levels as 3.0 mg selenium as sodium selenite, and 3.0 mg selenium from the yeast product increased the selenium concentration of whole blood by approximately equal to 40% and that of milk by approximately equal to 100%. The activity of glutathione peroxidase in erythrocytes of the group given selenite was not significantly different from that in either of the groups given the yeast product. The concentrations of selenium in the tissues of two cows from each group were marginal to adequate, and there was a trend for the concentrations to be higher in the tissues of the cows supplemented with the yeast product. PMID- 9342930 TI - Conceptual perspective and lexical choice in acquisition. AB - Adult speakers choose among perspectives when they talk, with different words picking out different perspectives (e.g., the dog, our pet, that animal). The many-perspectives account of lexical acquisition proposes that children learn to take alternative perspectives along with the words they acquire, and, therefore, from the first, readily apply multiple terms to the same objects or events. And adults offer children pragmatic directions about the meanings of new words and hence about new perspectives. In contrast, the one-perspective account proposes that children are able, at first, to use only one term to talk about an object or event. Evidence for the many-perspectives account comes from a range of sources: children spontaneously use more than one term for the same object (horse and chair for a toy horse); they construct novel words to mark alternate perspectives (Dalmation-dog vs. dog); they shift perspective when asked (from cat to animal, or sailor to bear for anthropomorphic characters); and they readily learn new terms for talking about already-labelled kinds. Children sometimes fail to learn new words or fail to relate them to words already known, but only in situations that lack adequate pragmatic directions. PMID- 9342931 TI - The representation of Hebrew words: evidence from the obligatory contour principle. AB - The Hebrew root morpheme typically consists of three consonants. Hebrew allows a gemination of a root consonant, but constrains its location [McCarthy, J. (1979). Formal problems in semitic phonology and morphology. Cambridge, MA; MIT Ph.D. dissertation. Distributed by Indiana University Linguistics Club. Garland Press, New York, 1985]. A gemination of a root-consonant is permitted at the end of the root (e.g., [mss]), but not at its beginning (e.g., [ssm]). Two experiments examined readers' sensitivity to the structure of the root morpheme by obtaining ratings for nonwords derived from nonroots. Root-initial gemination (e.g., [ssm]) was judged unacceptable compared to root-final gemination (e.g., [mss]) or no gemination controls (e.g., [psm]). The sensitivity to root structure emerged regardless of the position of the root in the word. These results have several implications. (1) Our findings demonstrate morphological decomposition. Hebrew speakers' ratings reflect a phonological constraint on the location of geminates. Being the domain of this constraint, the root morpheme must form a separate constituent in the representation of Hebrew words. (2) The rejection of root initial gemination supports the psychological reality of the Obligatory Contour Principle, a pivotal constraint in autosegmental phonology. (3) A sensitivity to the location of geminates presupposes a distinction between the representation of geminate and nongeminate bigrams. Such a distinction, however, requires the implementation of a symbol. Our findings converge with numerous linguistic evidence in suggesting that the representation of constituency structure is necessary to account for linguistic generalizations. PMID- 9342932 TI - Does rank have its privilege? Inductive inferences within folkbiological taxonomies. PMID- 9342933 TI - The functional anatomy of a hysterical paralysis. AB - The concept of a conversion disorder (such as hysterical paralysis) has always been controversial (Ron, M.A. (1996). Somatization and conversion disorders. In: B.S. Fogel, R.B. Schiffer & S.M. Rao (Eds.), Neuropsychiatry. Williams and Wilkins, Baltimore, MD). Although the diagnosis is recognised by current psychiatric taxonomies, many physicians still regard such disorders either as feigned or as failure to find the responsible organic cause for the patient's symptoms. We report a woman with left sided paralysis (and without somatosensory loss) in whom no organic disease or structural lesion could be found. By contrast, psychological trauma was associated with the onset and recurrent exacerbation of her hemiparalysis. We recorded brain activity when the patient prepared to move and tried to move her paralysed (left) leg and when she prepared to move and did move her good (right) leg. Preparing to move or moving her good leg, and also preparing to move her paralysed leg, activated motor and/or premotor areas previously described with movement preparation and execution. The attempt to move the paralysed leg failed to activate right primary motor cortex. Instead, the right orbito-frontal and right anterior cingulate cortex were significantly activated. We suggest that these two areas inhibit prefrontal (willed) effects on the right primary motor cortex when the patient tries to move her left leg. PMID- 9342934 TI - Polydnaviruses of hymenopteran endoparasitoids knock out the host insect immune response. AB - The insect immune system reacts against invading microorganisms and parasites with the recruitment of haemocytes and with humoral response. Cellular immune reactions involve phagocytosis, nodule formation and encapsulation by different types of haemocytes whereas insect cell-free antibacterial immunity depends on the production of a number of peptides and proteins, among which lysozyme, cecropins and attacins represent the major group of immune proteins. Polydnaviruses from certain hymenopterous parasitoids interfere with both host immunity and host development. These immunosuppressive viruses exhibit an intimate genetic relationship with the parasitoid since viral sequences are integrated within the parasitoid chromosomal DNA. The viral genes expression in parasitized host induces immunosuppression and alters development of the host insect. The parasitoids developing in the host body cavity knock out the insect immune system, inducing a decline in cellular and humoral components of the immune system so that parasitoid eggs are not recognized as foreign and thereby are not encapsulated. Polydnaviruses carrying parasitoids escape the host immune response and may develop within the insect host whereas other invaders are normally destroyed by defense factors of insect haemolymph. PMID- 9342936 TI - Effect of increased intracranial pressure on arterial peripheral and cerebral blood pressure. AB - The effect of increased intracranial pressure (ICP) on arterial peripheral (PBP) and cerebral blood pressure (CBP) was studied in 16 rabbits (New Zealand White). Changes of arterial blood pressure in peripheral and cerebral blood circulation were caused by a temporary (3 min) ICP increase. A raised ICP was obtained by infusing into the left lateral brain ventricle artificial cerebrospinal fluid at a pressure equal to 50, 100, and 150% of the initial CBP value. The experiments carried out showed that on reaching the initial CBP value, increased ICP induced Cushing response. The magnitude and duration of this reaction changed with increasing ICP, the intensity of the changes depending on the brain region in which the ICP increase was initiated. PMID- 9342935 TI - Preovulatory dynamics of ovarian steroid hormones in two mouse strains differing in the rate of meiotic maturation. AB - Immature females of two inbred strains, KE and CBA, differing in the rate of meiotic maturation, were used in this study. On the 26th day they were treated with 5 IU PMSG (0 h) and 48 h later with 5 IU hCG. Groups of females were killed at 2 h intervals starting with 46 h and ovaries were then dissected for steroid analysis by appropriate RIAs. In both strains hCG injection resulted in a sharp increase in ovarian progesterone content with a simultaneous fall in androgen and oestradiol concentrations. However, in the CBA strain, progesterone concentration started to rise earlier while androgen concentration decreased later than in the KE strain. This subtle difference could be one of the factors contributing to the different maturation rate of KE and CBA strains. PMID- 9342938 TI - Absence of acetylcholinesterase-positive nerve structure in the superficial part of the pineal gland of adult albino rats. AB - Application of the histochemical method for testing acetylcholiesterase (AChE, EC 3.1.1.7) combined with silver impregnation of the nervous tissue showed the absence of AChE-positive nerve structures in the superficial part of the pineal gland in adult male albino rats. It was carried out at the level of the light microscope. If it is assumed that the test for the presence of AChE is a form of indirect conclusion concerning the cholinergic nature of given nerve structures, then in the case of the superficial innervation of the pineal gland, the outcome of the test is negative. PMID- 9342937 TI - Effect of electrical stimulation of A and C fibres of the aortic nerves on arterial peripheral and cerebral blood pressure. AB - The effect of electrical stimulation of A and C fibres of the aortic nerves on arterial peripheral (PBP) and cerebral blood pressure (CBP) was studied in 12 rabbits (New Zealand White). The experiments have shown that selective stimulation of fibres A of the aortic nerves evokes every time depression of the arterial blood pressure in the peripheral circulation and slightly modifies pressure in the cerebral circulation. Selective stimulation of fibres C of the aortic nerves always elicits a significant decrease in arterial blood pressure in the peripheral circulation, whereas in the cerebral circulation it elicits a small decrease or a slight increase in arterial pressure. The obtained results point to a predominating role in depressor reaction of impulsation reaching the aortic arch through amyelic fibres C. The depressor reaction in the peripheral circulation is highest with simultaneous stimulation of the right and left aortic nerve during stimulation of both fibre A and C. PMID- 9342939 TI - Modulation of protein kinase C activity in NIH 3T3 cells by plant glycosides from Panax ginseng. AB - The involvement of ginsenosides in the signal cascade that stimulates cellular growth was investigated. It was found that ginsenosides Rh1 and Rh2 extracted from the root of Panax ginseng inhibited cellular proliferation in NIH 3T3 fibroblasts. Both ginsenosides Rh1 and Rh2 effectively reduced phospholipase C activity resulting in a decrease in the intracellular level of diacylglycerol, an endogenous activator of protein kinase C. The treatment of cells with Rh1 or Rh2 was thus found to reduce intracellular protein kinase C activity. We also observed that the phosphorylation of myristoylated alanine-rich C kinase substrate, one of the major substrates of protein kinase C in cells, was inhibited by the ginsenosides. Data suggest that the ginsenoside Rh1 or Rh2 exerts antiproliferative effects by inhibiting phospholipase C, which produces second messengers necessary for the activation of protein kinase C. PMID- 9342941 TI - Effects of brazilin on the altered immune functions in the early phase of halothane intoxication of C57BL/6 mice. AB - To investigate the effects of brazilin on the altered immune functions in the early phase of halothane intoxication in mice, several immune functions were investigated. Halothane was found to alter the immune functions which lead to hepatitis by autoimmune-mediated process. Based on the fact that immunomodulation at an initial step of autoimmune diseases is effective to prevent or control the diseases, in the present study the effects of brazilin on the altered immune functions in the early phase of halothane intoxication of C57BL/6 mice were investigated. By the treatment of halothane, delayed type hypersensitivity (DTH) and mitogen (ConA, LPS) induced proliferation of splenocytes were significantly increased and suppressor cell activity and mixed lymphocyte reaction (MLR) were decreased in C57BL/6 mice. But IgM plaque forming cells (PFCs) were not significantly changed. All the parameters tested were changed in homing patterns by the treatment with brazilin. But brazilin significantly increased IgM PFCs to higher than the normal level. PMID- 9342940 TI - Fractionation and chemical properties of immunomodulating polysaccharides from roots of Dipsacus asperoides. AB - A crude polysaccharide fraction (DAP-1) was prepared from roots of Dipsacus asperoides by hot water extraction and EtOH precipitation, and tested for anti complementary activity, mitogenic activity of lymphocytes, and effects on acid phosphatase and phagocytic activities of macrophages. DAP-1 showed not only anti complementary activity but also a stimulating effect on the mitogenic activity of lymphocytes. DAP-1 also significantly suppressed the phagocytic activity of macrophages. Although DAP-1 directly stimulated the mitogenecity of lymphocytes, it had no effect on lipopolysaccharide- or concanavalin A-induced mitogenic activity of lymphocytes. Periodate oxidation and pronase digestion suggested that the polysaccharide moiety in DAP-1 contributed to the expression of its anti complementary and mitogenic activities and that the protein moiety in DAP-1 was responsible for its effect on phagocytosis. DAP-1 gave three polysaccharide fractions (DAP-2, 3, and 4) by fractionation using cetyltrimethylammonium bromide. All the fractions had potent anti-complementary activity, but showed different stimulating or suppressive effects on the proliferation of lymphocytes, phagocytosis, or acid phosphatase activity. Three potent anti-complementary polysaccharides (DAP-4I-1a, DAP-4I-1b, and DAP-4IIa-1) were purified from DAP-4 by anion-exchange chromatography, gel filtration, and HPLC. DAP-4I-1a, I-1b, and IIa-1 consisted of Ara, Rha, Xyl, Gal, Glc and GlcA in a molar ratio of 1.0:0.7:1.0:18.6:22.2:nil; 1.0:0.1:0.3:19.3:26.8:nil; and 3.7:trace:0.6:26.3:5.5:1.0; respectively. Among the polysaccharides, only DAP 4IIa-1 reacted with beta-glucosyl-Yariv antigen. Methylation analysis indicated that DAP-4I-1a mainly comprised 4-linked Gal and 3-, 4-, and 6-linked Glc, whereas DAP-4IIa-1 consisted mainly of terminal Araf, 3-linked Glc, and 3,6 branched Gal. PMID- 9342942 TI - Brazilin modulates immune function mainly by augmenting T cell activity in halothane administered mice. AB - Previously we reported that brazilin, the main principle of Caesalpinia sappan, was able to improve the altered immune functions caused by halothane administration in mice. To elucidate the mechanisms of its immunomodulating activities, the effects of brazilin on the functions of T cells and splenic cellularity were investigated. Brazilin decreased splenic cellularity and IL-2 production which had been augmented in mice treated with halothane (21.5% in olive oil, 10 mmol/kg) for 4 consecutive days whereas the reduced expression of IL-2 receptors by ConA or standard IL-2 was increased by brazilin treatment. These data indicate that halothane induced a dysfunction of T cells resulting in abnormal immune responses and these altered immune functions might be improved mainly by affecting the function of T cells. PMID- 9342943 TI - Rutoside as mucosal protective in acetic acid-induced rat colitis. AB - The effect of the flavonoid rutoside on acetic acid-induced rat colitis was studied. Rats were pretreated orally with different doses of the flavonoid (10, 25, and 100 mg/kg) 48, 24, and 1 hour prior to colitis induction and examined for colonic damage 24 hours later. Colonic inflammation was characterized by gross and microscopical injury, bowel wall thickening, abolition of fluid absorption, glutathione depletion, enhanced leukotriene B4 synthesis, and increased levels of myeloperoxidase and alkaline phosphatase activities. Rutoside treatment (25 and 100 mg/kg) reduced histologic injury and prevented the increase in alkaline phosphatase activity, but it had no effect on myeloperoxidase levels or leukotriene B4 synthesis. In addition, glutathione depletion was effectively counteracted at the dose of 25 mg/kg, whereas fluid absorption was achieved at the highest dose assayed. It is concluded that rutoside has an acute anti inflammatory activity in this model which may be related to a putative direct protective effect on intestinal cells, mainly enterocytes, in which the antioxidative properties of the flavonoid may play a role. PMID- 9342944 TI - Protection of rat liver microsomes against carbon tetrachloride-induced lipid peroxidation by red ginseng saponin through cytochrome P450 inhibition. AB - A possible role of cytochrome P450 (P450) inhibition by red ginseng saponins in carbon tetrachloride (CCl4)-induced lipid peroxidation was investigated in liver microsomes prepared from male Sprague Dawley rats. The total saponin of red ginseng standardized on ginsenosides-Rb1, -Rb2, -Rc, -Rd, -Re, and -Rg1 whose composition was studied in our previous report was used in the present study. The standardized saponin of red ginseng showed inhibitory effects on P450-associated monooxygenase activities in a dose-dependent manner, particularly p-nitrophenol hydroxylase activity which has been known to represent CCl4-activating P450 2E1 enzyme. Meanwhile, silymarin enhanced the activity of P450 2E1 enzyme in liver microsomes. When the lipid peroxidation was induced by incubating rat liver microsomes with CCl4 in the presence of NADPH, the standardized saponin significantly blocked the formation of thiobarbituric acid-reactive substances at the same concentrations showing P450 inhibition in liver microsomes. Silymarin revealed more potent protection against CCl4-induced lipid peroxidation. When the lipid peroxidation was induced by FeCl3, in which metabolic activation may not be required, only silymarin could protect the lipid peroxidation in liver microsomes. Taken together, our present results indicated that the inhibitory effects of red ginseng saponin on P450 enzymes may have a critical role in CCl4 induced lipid peroxidation in rat liver microsomes and that the mechanism of hepatoprotection by red ginseng saponin may be distinct from that of silymarin. PMID- 9342945 TI - Stimulative effects of saponin from kikyo-to, a Japanese herbal medicine, on pancreatic exocrine secretion of conscious rats. AB - Our previous report stated that kikyo-to, a Japanese herbal medicine, consisting of the roots of Platycodon grandiflorum and Glycyrrhiza sp., stimulates the pancreatic exocrine secretion of conscious rats. The present study focused on the effective components of kikyo-to and the mechanism of stimuli to pancreatic secretion of rats. When 10 to 100 mg of platycodin D, a saponin from the root of Platycodon grandiflorum, was intragastrically administered, the pancreatic secretion of rats was stimulated. At the same time, the plasma CCK concentration increased. On the other hand, the stimulative effects of glycyrrhizin, a saponin from the root of Glycyrrhiza sp. were weak compared to platycodin D. The effects of 10 mg/kg of platycodin D on pancreatic secretion were inhibited by loxiglumide (50 mg/kg, i.g.), a CCK receptor antagonist. In contrast, the suppressive effect of atropine (300 micrograms/kg/h, i.v.) on pancreatic secretion was reduced by administering 10 mg/kg of platycodin D. In addition, up to 1 mM of platycodin D did not inhibit the trypsin activities in vitro. In conclusion, kikyo-to serves to stimulate pancreatic exocrine secretion mainly because platycodin D causes gastrointestinal hormones, particularly, CCK to be released from the duodenum. PMID- 9342946 TI - Bisbenzylisoquinoline alkaloids from Stephania cepharantha and their effects on proliferation of cultured cells from the murine hair apparatus. AB - Bisbenzylisoquinoline alkaloids were isolated from Stephania cepharantha Hayata and their proliferative activities on cultured hair cells from the murine skin were evaluated. Cepharanthine (1), cepharanoline (9), isotetrandrine (2), and berbamine (7) showed significant activities in the range of 0.01-0.1 microgram/ml, but had no activity on cultured keratinocytes or fibroblasts from the murine skin. PMID- 9342947 TI - Antiviral activity of natural sulphated galactans on herpes virus multiplication in cell culture. AB - A sulphated galactan (SG) with low molecular weight (app. 2800) was isolated from extracts of Cryptopleura ramosa, a red seaweed from the South American coasts. The compound was a selective inhibitor of HSV-1 and HSV-2 replication in Vero cells with 50% inhibitory concentrations (IC50) in the range 1.6-4.2 micrograms/ml and a 50% cytotoxic concentration (CC50) of 476 micrograms/ml. SG was also effective against HSV-1 in cells of neural origin such as murine astrocytes. The mode of action of SG could be ascribed to an inhibitory action on virus adsorption. Furthermore, SG did not inhibit the blood coagulation process at concentrations highly exceeding the IC50. PMID- 9342949 TI - Role of human intestinal Prevotella oris in hydrolyzing ginseng saponins. AB - The potential of intestinal bacteria to hydrolyze ginsenoside Rb1 to 20-O-beta-D glucopyranosyl-20(S)-protopanaxadiol (I) was found in 79% of the fecal specimens from 58 human subjects whose age ranged from 1 to 64 years. Following a ginsenoside-Rb1-hydrolyzing activity assay, Prevotella oris strains were then isolated as a major bacterial species possessing the potential. All the intestinal isolates converted ginsenosides Rb1 and Rd to I, ginsenoside Rb2 to 20 O-[alpha-L-arabinopyranosyl(1-->6)-beta-D-glucopyranosyl]-20(S) -protopanaxadiol (II), and ginsenoside Rc to 20-O-[alpha-L-arabinofuranosyl(1-->6)-beta-D glucopyranosyl]-20(S)- protopanaxadiol (III) like fecal microflora, but did not attack ginsenosides Re or Rg1 (protopanaxatriol-type). The isolates were susceptible to colimycin (MIC, 3.13 micrograms/ml) and then the treatment of specific pathogen free mice with colimycin (20 mg/kg/day) decreased intestinal bacterial Rb1-hydrolyzing potential from 22.1 +/- 1.2% to 4.7 +/- 2.7%, while the decreased potential was restored to 30.7 +/- 3.7% by the inoculation with P. oris isolates. These results suggest that the metabolism of protopanaxadiol saponins to metabolites I-III in the intestines seems most partly due to intestinal P.oris. In addition, the fact that neither intact ginsenoside Rb1 nor its middle metabolic derivatives but only the final metabolite I was detected at 1.0-7.3 micrograms/ml in blood after oral administration of mice with ginsenoside Rb1 (125 mg/kg) allows us to speculate that metabolites I-III are the most likely forms of protopanaxadiol saponins absorbed from the intestines. PMID- 9342948 TI - Experimental treatment of cutaneous leishmaniasis with argentilactone isolated from Annona haematantha. AB - From the hexanic extract of roots of Annona haematantha an alpha,beta-unsaturated delta-lactone was isolated and identified as argentilactone. This compound exhibited in vitro activity against various strains of Leishmania ssp. at 10 micrograms/ml. BALB/c mice infected with Leishmania amazonensis were treated four weeks after infection with argentilactone by oral or subcutaneous routes for 14 days at 25 mg/kg daily. The reference drug, N-methylglucamine antimonate, was administered by subcutaneous injections at 100 mg/kg for 14 days. In these conditions, argentilactone showed the same efficacy as the reference drug, reducing by 96% the parasite loads in the lesion and by 50% the parasite burden in spleen. PMID- 9342950 TI - Extracellular polysaccharides produced by suspension-cultured cells from Digitalis lanata. AB - Extracellular polysaccharides (ECP) were isolated in yields of up to 4 mg/ml from the culture media of suspension-cultured cells from Digitalis lanata Ehrh. ECP content was increasing continuously over the first ten days of cultivation and then stayed constant until day 20. ECP were fractionated by ion-exchange chromatography into two neutral and one acidic fractions. Further fractionation was achieved by gel-permeation chromatography (GPC). One neutral fraction was separated into two distinct fractions with average molecular weights of 160 and 70 kDa, respectively. The second neutral fraction was hetero-disperse in GPC with average molecular masses of 112, 32, and 8 kDa. Polysaccharides of all neutral fractions consisted of glucose, xylose, galactose, and arabinose. Methylation analysis indicated these fractions to contain xyloglucans besides minor amounts of highly branched arabinogalactans. Xyloglucans were, using endo-beta-(1- >4)glucanase, fragmented into subunits which were identified mainly as tri- and pentasaccharides. The acidic fraction eluated as a single peak during gel permeation chromatography with an average molecular weight of 56 kDa. Analysis of carbohydrate composition and linkage analysis indicated that this polysaccharide is an acidic arabinogalactan. 2,6-Dideoxysugars, the typical carbohydrate components of cardiac glycosides in Digitalis lanata, were not detected in ECP. PMID- 9342951 TI - Characterization and cytokine-stimulating activities of acidic heteroglycans from Tremella fuciformis. AB - Four acidic heteroglycans, T2a-T2d, were isolated from the body of Tremella fuciformis Berk. They contained 1.9%-2.9% of acetyl groups and were composed of mannose (Man), glucuronic acid (GlcA), and small amounts of xylose (Xyl), glucose (Glc), and fucose (Fuc). According to methylation analysis they had a mannan backbone consisting of 3-linked Man, and side chains containing glucosyl, mannosyl, fucosyl, xylosyl, and glucuronic acid residues. The side chains were attached through O-2, O-4, or O-6 in about 40 percent of backbone mannosyl residues. Molecular masses of the four polysaccharides were 410, 250, 34, and 20 kDa, respectively. T2a-T2d induced human monocytes to produce interleukin-1 (IL 1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) in vitro. The products of Smith degradation (T2a-S) and lithium degradation (T2a-L) of T2a and the product of deacetylation (T2b-D) of T2b also induced monocytes to secret IL-1 as efficiently as the original polysaccharides, indicating that xylosyl and glucuronic acid residues as well as acetyl groups were not important to promote the cytokine-stimulating activity. PMID- 9342952 TI - Lack of anticholinergic activity by baicalin in the isolated trachea of guinea pig. AB - The anticholinergic effect of baicalin on the trachea from asthmatic guinea pig had been described elsewhere. Using 1 x 10(-5) M bethanechol to induce contraction in the isolated trachea from non-asthmatic guinea pig, baicalin below 1 x 10(-4) M was unable to attenuate the bethanechol-induced tracheal contraction; however, 1 x 10(-4) M baicalin was demonstrated to increase the tracheal contraction in the presence of bethanechol. Baicalin (1 x 10(-4) M) was also able to induce tracheal contraction in the absence of bethanechol, and this baicalin-induced tracheal contraction was shown to be atropine-sensitive. These data suggested that the baicalin effect on the isolated trachea from non asthmatic guinea pig was not anticholinergic in nature. PMID- 9342953 TI - Oral administration of a unicellular green algae, Chlorella vulgaris, prevents stress-induced ulcer. AB - Oral administration of dry powder of Chlorella vulgaris (CVP) showed clear prophylactic effects in water-immersion restraint stress-induced and in cysteamine-induced peptic ulcer models, but not in Shay's rat model. Drugs that enhance the protective factors of ulcer formation are effective in the first two models. CVP may prevent ulcer formation mainly through the "immune-brain-gut" axis and protection of gastric mucosa by its own characteristics. PMID- 9342954 TI - Evaluation of four Narcissus cultivars as potential sources for galanthamine production. AB - Galanthamine, an alkaloid present in the Amaryllidaceae is currently undergoing clinical trials for the treatment of Alzheimer's. Common daffodils, Narcissus spp., contain galanthamine and other alkaloids. Four commercial Narcissus cultivars were evaluated as potential sources of galanthamine. Planting depths, planting densities, bulb size or flower bud removal did not affect galanthamine content. PMID- 9342955 TI - Artekeiskeanin A: a new coumarin-monoterpene ether from Artemisia keiskeana. AB - A new coumarin-monoterpene ether, artekeiskeanin A (1) and two known coumarins, dracunculin (2) and scopoletin (3) were isolated from the aerial parts of Artemisia keiskeana. The structure of 1 was determined to be 7-(trans-8 oxogeranyloxy)-6-methoxycoumarin on the basis of spectroscopic studies. PMID- 9342956 TI - Composition and antimicrobial activity of the essential oil of Cistus creticus subsp. eriocephalus. AB - The chemical composition of the essential oil of the leaves of Cistus creticus subsp. eriocephalus (Viv.) Greuter & Burdet, (Cistaceae), was investigated by GC/MS. Thirty-nine components were identified, representing 73.9% (based on % total peak area by GC) of the oil composition. The main components of the oil were alpha-cadinene (6.5%), delta-cadinene (5.6%), viridiflorol (5.4%), bulnesol (6.3%), ledol (3.8%), alpha-copaene (3.8%), beta-selinene (3.4%), cubenene (3.3%), manoyl oxide (9.9%) and 13-epi-manoyl oxide (3.4%). Antibacterial studies were carried out in vitro against Gram-positive and Gram-negative organisms. PMID- 9342957 TI - Phytochemical analysis of Geigeria alata and Francoeuria crispa essential oils. AB - Phytochemical analyses of Geigeria alata (Benth. & Hook.) and Francoeuria crispa (Forssk., Cas.) (Asteraceae) essential oils were performed. G. alata oil showed moderate in vitro cytotoxicity (IC50, micrograms/ml against tumor cells; P388: 2.0, A-549: 2.5 and HT-29: 5.0), and also showed weak anti-HIV activity. S Carvotanacetone, the major component of F. crispa oil (93.0%), was isolated and its structure was elucidated by 2D-NMR analysis. PMID- 9342958 TI - The dimensionality of the flanker compatibility effect: a psychophysiological analysis. AB - The psychophysiological approach was used to evaluate the effects of feature similarity and "intrinsic response mapping" on the flanker compatibility effect. Symbol (e.g., < > < and 1 category in 281 vs 62 of the patients (P < 0.0001); mobility and psychological distress indices also improved (P = 0.006, and P = 0.007). CONCLUSIONS: Once-daily nitrate therapy not only provides a better NYHA angina classification than multiple-dose therapy does, but also provides a better quality of life as estimated by improvement of mobility and distress indices, the most important indicators of quality of life in this category of patients. PMID- 9342965 TI - Quantitative angiographic analysis of PTCA-induced coronary vasoconstriction in single-vessel coronary artery disease. AB - Quantitative coronary angiography was applied to investigate the degree and extent of coronary vasoconstriction following percutaneous transluminal coronary angioplasty (PTCA) in single-vessel disease of segments distal to the PTCA site (n = 46) and of control segments in nonmanipulated vessels (n = 33) before PTCA, 15 minutes after PTCA, and again 10 minutes after 10 mg sublingual isosonbide dinitrate (ISDN) in 46 patients receiving neither nitrates nor calcium channel blockers prior to PTCA. Furthermore, the degree of coronary vasoconstriction was compared with ergonovine-induced vasoconstriction (n = 8) as well as in patients with and without restenosis during follow-up angiography 4 months later. PTCA induced a moderate, but significant, vasoconstriction in both distal and control segments, with a reduction in coronary diameter from 2.34 +/- 0.58 to 2.26 +/- 0.55 mm (P = 0.011) and from 2.70 +/- 0.62 to 2.60 +/- 0.65 mm (P = 0.004), respectively. No correlation between the degree of vasoconstriction on the one side and lesion severity and PTCA-induced mechanical stretch, judged by the sum of the products of inflation pressure and time, on the other side was found. Vasoconstriction was within the limits achievable with the potent vasoconstrictor ergonovine and did not differ in patients with or without restenosis. ISDN led to a significant vasodilatation in all segments. In conclusion, coronary vasoconstriction following PTCA is present in the coronary tree in a rather diffuse way. It is not associated with stenosis severity or PTCA-induced mechanical stretch, suggesting a complex underlying mechanism. ISDN-reversible vasoconstriction was within the limits achievable with ergonovine and did not differ with regard to restenosis. PMID- 9342966 TI - Left ventricular diastolic filling dynamics during isometric exertion in syndrome X assessed with Doppler flowmetry. AB - To study left ventricular diastolic properties in syndrome X, we analyzed transmitral filling dynamics during handgrip exertion. In 14 normal subjects (N), 17 with syndrome X (Syn X), 16 with single-vessel disease (SVD), and 8 with multiple-vessel disease (MVD), transmitral inflow was recorded at baseline and during handgrip (50% of maximal effort for one minute) using pulsed Doppler echocardiography. We measured early diastolic (E) and late atrial (A) inflow velocities, A/E ratio and percent change of A/E from baseline (%A/E). Blood pressure and heart rate increased to the same degree in each group during handgrip. In normal subjects, E did not change with handgrip; A (51 +/- 10 vs 54 +/- 11 cm/sec, P < 0.05) and A/E (1.16 +/- 0.22 vs 1.25 +/- 0.33, P < 0.05) increased minimally. In Syn X subjects, E decreased (51 +/- 10 vs 38 +/- 10 cm/sec, P < 0.0001), A increased (52 +/- 11 vs 60 +/- 14 cm/sec, P < 0.005), and A/E increased markedly (1.07 +/- 0.31 vs 1.68 +/- 0.51, P < 0.0001). The %A/E in Syn X and MVD were significantly larger than that in SVD and N (Syn X: 58 +/- 29%; MVD: 45 +/- 25%; SVD: 22 +/- 21%; N: 8 +/- 13%). Handgrip-induced changes in diastolic filling in syndrome X and are similar to those in MVD and more marked than in SVD. These changes are consistent with impaired ventricular relaxation and support a generalized left ventricular (LV) abnormality in syndrome X. PMID- 9342968 TI - Relationship between ultrasound assessment of arterial wall properties and blood pressure. AB - Physical properties of an artery can be described in terms of stiffness, distensibility, and compliance. Changes in these properties can predict atherosclerosis disease, but it is necessary to identify an index that is independent of changes in blood pressure. We measured common carotid artery diameter and pulsatile change with an ultrasonic phase-locked, echo-tracking system in 7 subjects whose mean brachial blood pressure had varied 15 mm Hg or more within a month. Patients taking antihypertensive agents were excluded from the study. We measured changes in arterial diameter (n = 41) at least four times during the study period and calculated the pressure-strain elastic modulus (Ep), distensibility coefficient (DC), cross-sectional compliance (CC) and stiffness parameter (beta) from inner diameter, its pulsatile change, and blood pressure. The correlation coefficients of mean blood pressure with each index are as follows: Ep, 0.53; DC, 0.58; CC, 0.63; beta, 0.21. When mean blood pressure increased 1 mm Hg, the change in each index at 100 mm Hg was as follows: Ep, 1.48 +/- 1.30%; DC, -1.05 +/- 0.97%; CC, -0.69 +/- 0.90%; beta, 0.45 +/- 1.11%. Among the four indices that measure the properties of the arterial walls, stiffness parameter beta was the least dependent on blood pressure. Thus, it appears to have usefulness as an index of arterial wall sclerosis. PMID- 9342967 TI - Physical activity is related to ankle/brachial index in subjects without peripheral arterial occlusive disease. AB - The purpose of this study was to determine whether physical activity level was directly and independently related to the ankle/brachial systolic blood pressure index (ABI) in subjects without peripheral arterial occlusive disease (PAOD). A total of 353 subjects between the age of 38 and 88 years (63.7 +/- 9.1 years; mean +/- standard deviation) who had ABI values > or = 1.00 were studied. The sample consisted of 230 men and 123 women, of whom 274 were caucasian and 79 were African-American. Subjects were characterized on blood pressure, physical activity level from validated questionnaires, anthropometry, plasma lipoprotein lipids, and smoking history. The ABI (1.16 +/- 0.13) was related to physical activity obtained from the Minnesota Leisure Time Physical Activity (LTPA) questionnaire (r = 0.413, P < 0.001). Multiple regression analysis identified race, hypertension, current smoking status, and body mass index (BMI) as cardiovascular risk factors that were independently related to ABI. After controlling for these variables, the relationship between ABI and physical activity persisted (partial r = 0.329, P < 0.001). Thus, physical activity was positively related to ABI in subjects free of PAOD, and the relationship between physical activity and ABI persisted after controlling for race, hypertension, current smoking status, and BMI. It is concluded that adopting a physically active lifestyle is associated with a reduced risk of developing PAOD. PMID- 9342969 TI - Prethrombotic state due to hypercoagulability in patients with permanent transvenous pacemakers. AB - Venous thrombosis is a relatively usual but serious complication of permanent transvenous pacing. However, the pathogenesis has not been defined. To clarify underlying abnormalities in the coagulation-fibrinolysis system in patients with permanent transvenous pacemakers, we measured serum levels of fibrinopeptide A (FPA), thrombin-antithrombin III complexes (TATs), plasmin-alpha 2 plasmin inhibitor complexes (PICs), D-dimer (D-D), beta-thromboglobulin (beta-TG), and platelet factor 4 (PF4) in 53 patients with permanent transvenous pacemakers and 10 control subjects. The patients were divided into two groups, as follows, according to the presence of mural thrombus documented along the pacing lead(s) by digital subtraction angiography and transesophageal echocardiography: Group Th (-), patients without venous route thrombus; and Group Th (+), patients with venous route thrombus. FPA and TAT levels increased significantly even in Group Th (-), and further increased in Group Th (+) compared with control subjects (FPA: 7.5 +/- 4.9, 15.3 +/- 8.8 vs 3.0 +/- 1.4 ng/mL, respectively, P < 0.05; TAT: 2.9 +/- 1.3, 4.8 +/- 2.3 vs 1.7 +/- 0.6 ng/mL, respectively, P < 0.05). There were no differences in levels of D-D, PIG, beta-TG, and PF4 among control subjects, Group Th (-), and Group Th (+). These findings suggest that the hypercoagulable state appears in patients with permanent transvenous pacemakers, even without apparent venous thrombosis. The patients with permanent transvenous pacemakers are thought to be in the prethrombotic state even if they have no venous route thrombosis. PMID- 9342970 TI - Coronary wallstent implantation by the transradial artery approach. A case report. AB - The authors report the implantation of two wallstents in a patient by use of the transradial artery approach. This approach for coronary wallstent implantation allows for intensive anticoagulation therapy with less risk of bleeding. PMID- 9342971 TI - New stenosis on proximal coronary segment after directional atherectomy. Two case reports. AB - Two patients with new coronary stenotic lesions subsequently developed proximal to the sites accepting directional coronary atherectomy (DCA) are reported. One lesion developed at the left main coronary artery and the other at the proximal segment of the left anterior descending artery. The mechanisms of the development of such new stenotic lesions after DCA were studied and procedure-related mechanical trauma over the proximal segment of the primary lesion may be the possible mechanism for such complication. PMID- 9342972 TI - Right ventricular electrode lead implantation via a persistent left superior vena cava. An improved technique. AB - Persistent left superior vena cava occurs in approximately 0.5% of the population. This may complicate pacemaker implantation by making lead insertion difficult and causing lead instability through the left cephalic vein and the subclavian vein approach. We used a wide loop technique in the right atrium and successfully advanced the lead in the right ventricle apex. A persistent left superior vena cava does not preclude successful lead placement. PMID- 9342973 TI - Unusual bleeding complications of thrombolytic therapy after cardiopulmonary resuscitation. Three case reports. AB - The authors present three case reports retrospectively casting doubt on the benefit of thrombolysis after external cardiac massage. PMID- 9342974 TI - Cardiac contusion--a diagnostic dilemma. PMID- 9342975 TI - Nitroglycerin echocardiography and dobutamine. PMID- 9342976 TI - A syndrome of ear and sinus symptoms dependent upon disturbed function of the temporomandibular joint. 1934. PMID- 9342977 TI - Jaws revisited: Costen's syndrome. AB - Although James Costen was not the first to ascribe ear pain, tinnitus, impaired hearing, and even dizziness to temporomandibular joint dysfunction, he developed an integrated and systematic approach ascribing the symptoms to dental malocclusion. He wrote extensively on it, and a few years after his original article, the term Costen's syndrome came into general use. Recently, the use of the eponym has decreased, as dental malocclusion has assumed a lesser role in explaining many of the symptoms formerly ascribed to it. PMID- 9342978 TI - Meniere's syndrome and migraine: incidence in one family. AB - Since 1992, we have applied a standard questionnaire to all our Meniere's syndrome patients. We ask about other family members affected by the symptoms and about the presence of the usual migraine symptom. Through this questionnaire we have identified a family that has some members affected by Meniere's syndrome alone, some others with associated migraine, and still others with migraine alone. Two members of this family started out with migraine and later in life developed Meniere's syndrome. The genetic transmission follows an autosomal dominant pattern for both Meniere's syndrome and migraine. We have interviewed and studied 19 affected persons from several generations of this family, who form the basis of this report. The possibility of a common autosomal dominant genetic determinant for Meniere's syndrome and migraine and its implications for the causation of Meniere's disease are discussed. PMID- 9342979 TI - Perilymphatic fistula: a Washington, DC, experience. AB - One hundred ninety-seven patients who underwent surgical repair for a presumed unilateral perilymphatic fistula were reviewed. Of those patients, 87% with vestibular symptoms reported complete or near-complete relief of their symptoms. Forty percent of the patients with sudden hearing loss had an improvement in their hearing levels. An analysis of several diagnostic tests revealed their sensitivity and specificity ratings. A review of the patients' operative records showed a marked disparity between the visual identification of an actual fluid leak during surgery and their postoperative outcome. This review supports the premise that at the present time, the patient's surgical outcome is the best way of documenting a successfully repaired perilymphatic fistula. PMID- 9342980 TI - Vestibular and optokinetic function in the normal guinea pig. AB - Vestibular and optokinetic function was quantitatively studied in the normal guinea pig through investigation of the vestibulo-ocular reflex (VOR), optokinetic nystagmus (OKN), and the visual vestibulo-ocular reflex (VVOR) by means of sinusoidal stimulation with a computer-controlled rate table (VOR and VVOR) or an optokinetic drum. The VOR exhibited high-pass filter characteristics with steady state gain achieved at 0.125 Hz. The maximum gain was 0.55 at a velocity of 60 degrees/s. The VOR was modeled by a transfer function with best fits obtained with an adaptation time constant of 12.5 seconds. The OKN showed low-pass filter characteristics with a decrease in gain for increase in stimulus amplitude. The maximum gain measured was 0.64. A fractional pole model provided a fit of these data. The VVOR exhibited a mean gain of between 0.6 and 0.7 across the stimulus bandwidth and peak velocities. A model based on a linear combination of the actual OKN and VOR gains provided an estimate of the VVOR gain. PMID- 9342981 TI - Effect of immunotherapy on serum levels of eosinophil cationic protein in perennial allergic rhinitis. AB - Eosinophil cationic protein (ECP) levels in the serum of clotted blood could reflect the rate of activation of circulating eosinophils. We investigated the serum ECP levels in patients with perennial allergic rhinitis, with special reference to the effect of immunotherapy on the serum ECP levels. Serum ECP levels in untreated patients with perennial allergic rhinitis are significantly higher than those of nonatopic volunteers. Therefore, this elevation in the untreated patients represents an ongoing inflammation occurring in allergic rhinitis. The mean serum ECP level of a 1-year immunotherapy group was significantly higher than that of the nonatopic group, and was not different from that of the untreated group. In contrast, the mean serum ECP level in patients who had more than 2 years of immunotherapy was significantly lower than that of the untreated group, and was not different from that of the nonatopic group. Additionally, serum ECP levels were significantly correlated with the duration of immunotherapy. These findings suggest that activation of circulating eosinophils decreases gradually during immunotherapy, but this inhibition becomes apparent only after 2 years of immunotherapy. The control of circulating eosinophil activation might be one of the important working mechanisms behind the clinical effect of immunotherapy. PMID- 9342982 TI - Incidence of primary ciliary dyskinesia in children with recurrent respiratory diseases. AB - The goal of the study was to evaluate the incidence of primary ciliary dyskinesia (PCD) in children suffering from recurrent respiratory tract infections (RRIs) by means of noninvasive method. Respiratory ciliated cells were collected by nasal brushing in 118 children (4.6 +/- 2.5 years) with RRIs. The ciliary beat frequency (CBF) was measured with a stroboscopic method, and when the CBF was abnormal, the ciliary ultrastructure was analyzed by a quantitative method. The CBF could be measured in 106 patients (90%) and was abnormal in 15 patients. The ciliary ultrastructure was found to be abnormal in 11 of 15 patients: PCD was diagnosed in 6 cases, and acquired ciliary defects were observed in the remaining 5 patients. Our conclusion, that PCD is rare but net exceptional (5.6%) in children with RRIs, justifies the systematic investigation of ciliated cells in such patients. For this purpose, nasal brushing can be used to sample ciliated cells even in young children. PMID- 9342983 TI - In vivo measurement of human nasal mucociliary motility using a laser light scattering instrument. AB - The mucociliary system is one of the most important airway defense mechanisms, and knowledge of the mucociliary wave frequency (MWF) is important in the understanding of this system. Employing a laser light scattering technique and a thin, flexible fiberoptic probe, we developed and tested a simple and practical device for real-time in vivo measurements of mucociliary activity in the human nose. The laser instrument is user-friendly and does not produce any discomfort to the patient. The mean +/- SE of MWF of 36 measurements in 16 normal subjects was 7.7 +/- 0.5 Hz. The mean MWF of 17 measurements in 7 patients with allergic rhinitis was 5.5 +/- 0.2 Hz (p < .005), and the mean MWF of 56 measurements in 17 patients with septum deviation was 5.8 +/- 0.2 Hz (p < .001). The instrument presented in this study might provide a new and convenient method of studying the mucociliary activity in the respiratory tract. PMID- 9342984 TI - Effects of anesthetic technique on the hemodynamic response to microlaryngeal surgery. AB - Our aim was to study the attenuation of the hemodynamic response to microlaryngeal surgery by beta-blocking agents used as support drugs to the anesthetic technique. The study was carried out in 30 patients randomly allocated to one of three groups. The control group received only anesthetic drugs. The second group received labetalol hydrochloride 0.3 mg/kg 3 minutes before induction and 0.15 mg/ kg 2 minutes prior to the start of suspension of the larynx. The third group received esmolol hydrochloride 500 micrograms/kg 3 minutes prior to induction and a continuous infusion of 300 micrograms/kg during the surgical procedure. Hemodynamic data in the three groups were compared by analysis of variance. A statistically significant difference (p < .05) was found in hemodynamic data among the two groups treated with blocking agents and the control group. The addition of beta-blocking agents to the anesthetic technique attenuates the hemodynamic response to suspension laryngoscopy. PMID- 9342985 TI - QT interval changes following neck dissection. A stratified prospective study. AB - Studies from Europe have suggested that neck dissection, especially right radical neck dissection, causes a dangerous prolongation of the QT interval. Sudden cardiac arrest due to QT prolongation has been reported following right radical neck dissection. We investigated the prevalence of QT interval prolongation following neck dissection. Electrocardiogram tracings from 45 patients who underwent different combinations of neck dissection were studied. Preoperative and postoperative tracings were interpreted by a cardiologist blinded to the patient identification of each tracing. There were 28 unilateral neck dissection patients and 17 bilateral neck dissection patients eligible for analysis. There were 7 patients in the classic right radical neck dissection group, and only 3 of these had no neck dissection on the left. Comparisons of preoperative versus postoperative corrected QT interval for all subjects did not indicate a significant change. Stratification by neck dissection type (radical, modified or selective, and carotid artery resection) or by side dissected (left, right, or both) also showed no significant differences. No malignant arrhythmias were encountered. Thus, in contrast to the European experience, our findings show no significant predictable change in the QT interval after any of the combinations of neck dissection. Head and neck surgeons should be aware of the possibility of postoperative QT interval prolongation following neck dissection, although in the absence of other risk factors it appears to be a rare occurrence. PMID- 9342986 TI - Intracranial complications of otitis media. PMID- 9342987 TI - Calcified Mucor fungus ball of sphenoid sinus: an unusual presentation of sinoorbital mucormycosis. PMID- 9342988 TI - Syndrome of inappropriate antidiuretic hormone secretion associated with head neck cancers: review of the literature. AB - In a minority of patients with malignant tumors, signs and symptoms develop that cannot be explained on the basis of the mass effect produced by the primary tumor or its metastases, or production of a hormone normally associated with the tissue type that has given rise to the malignant tumor; these peculiar symptom complexes are known as paraneoplastic syndromes, and may be divided into endocrinologic, dermatologic, hematologic, neurologic, and osteoarticular manifestations. In the head and neck region in particular, the syndrome of inappropriate antidiuretic hormone production (SIADH, or Schwartz-Bartter syndrome) is a well-recognized form of paraneoplastic syndrome that may accompany head and neck malignancies. Most of such tumors are squamous carcinomas, with lesser numbers of olfactory neuroblastomas, small cell neuroendocrine carcinomas, adenoid cystic carcinomas, and undifferentiated carcinomas; sarcoma was reported in only a single instance. The lesions associated with the development of SIADH have most often been located in the oral cavity, and less often in the larynx, nasopharynx, hypopharynx, nasal cavity, maxillary sinus, parapharyngeal space, salivary glands, and oropharynx. Key features of SIADH include serum hypo-osmolality; an unexpectedly high urinary specific gravity; an absence of edema or dehydration; normal adrenal, thyroid, and renal function; hyponatremia; and an elevation of plasma vasopressin. PMID- 9342989 TI - Nucleus 20-channel and 21-channel auditory brain stem implants. First European experiences. PMID- 9342990 TI - European auditory brain stem prosthesis. PMID- 9342991 TI - Criteria for selecting the side for cochlear implantation. PMID- 9342992 TI - Waveforms of cochlear implant-evoked auditory brain stem responses in anesthetized young children, recorded with a new preamplifier. AB - An ultra-low-noise preamplifier has been specifically designed for the conditions in lightly anesthetized young children, to record cochlear implant--evoked auditory brain stem responses (impEABRs). The preamplifier linearly filters out the signal from the radio-frequency implant-driver. The preamplifier provides simultaneous accurate records of the stimulus-current artifact and impEABR, both ipsilaterally and contralaterally. It is convenient to label electrically evoked vertex-positive peaks, eg, eII, on probable analogy with acoustically evoked potentials. In order to label all the commonly observed peaks with latency less than 5 milliseconds, it is necessary at times to use labels eIIa, eIIb, eIIIa, eIIIb, eIVa, eIVb, eVa, and eVb. The records also show two distinct nadirs, with latencies about 2.5 milliseconds and about 6 milliseconds. The details of the waveforms are preserved as the stimulus strength descends towards threshold. PMID- 9342993 TI - Duration of anticoagulant therapy after first episode of venous thrombosis in patients with inherited thrombophilia. PMID- 9342994 TI - Estrogen replacement therapy and mortality among older women. The study of osteoporotic fractures. AB - BACKGROUND: Most previous studies of estrogen replacement therapy (ERT) and mortality have focused on younger women. Recently, it has been suggested that the effect of ERT on mortality may represent a "healthy-user" effect, ie, those with healthier lifestyles having a greater likelihood of receiving ERT. METHODS: Nine thousand seven hundred four women, 65 years or older, participated; 1258 (14.1%) reported current use of ERT for at least 1 year at entry. During an average follow-up of 6.0 years, 1054 women (11.8%) died. RESULTS: After adjusting for multiple variables, mortality rate was lower among current (relative risk [RR], 0.69; 95% confidence interval [CI], 0.54-0.87) and past users (RR, 0.79; 95% CI, 0.66-0.95), mainly due to reductions in deaths due to cardiovascular disease. The protective effect of ERT was greatest among women younger than 75 years (RR, 0.55; 95% CI, 0.40-0.76) compared with women from 75 to 84 years of age (RR, 0.93; 95% CI, 0.62-1.41) and 85 years or older (RR, 1.33; 95% CI, 0.43-4.12). The RR for overall mortality was 0.95 (95% CI, 0.68-1.32) among short-term users (1-9 years) compared with 0.55 (95% CI, 0.40-0.75) among long-term users (> or = 10 years). Deaths considered unrelated to ERT tended also to be reduced in current users younger than 75 years (RR, 0.72; 95% CI, 0.49-1.06) and current long-term users (RR, 0.75; 95% CI, 0.51-1.10). CONCLUSIONS: Estrogen replacement therapy is associated with lower overall mortality rates and reduced deaths due to cardiovascular disease. Women using ERT had healthier lifestyles, and the risk for death thought to be unrelated to ERT also tended to be lower in ERT users, suggesting in part a healthy-user effect. PMID- 9342995 TI - Risk factors for 30-day mortality in elderly patients with lower respiratory tract infection. Community-based study. AB - BACKGROUND: Pneumonia is a major cause of death in the elderly, but there are few studies of risk factors for death that include both ambulatory and nursing home patients. OBJECTIVE: To assess factors associated with 30-day mortality in a population-based study of older adults with lower respiratory tract infection. METHODS: Identification of (1) a previously identified retrospective cohort of all residents of Rochester, Minn, aged 65 years or older who experienced a first episode of pneumonia or bronchitis during a calendar year and (2) the risk factors associated with 30-day mortality through review of complete inpatient and ambulatory medical records. Logistic regression was used to identify significant independent risk factors for 30-day mortality. RESULTS: A total of 413 adults aged 65 years or older were identified. The independent factors for 30-day mortality were atypical symptoms (odds ratio [OR], 4.98; 95% confidence interval [CI], 2.14-11.60), neurologic illness (OR, 3.92; 95% CI, 1.47-6.59), current diagnosis of cancer (OR, 6.2; 95% CI, 2.40-15.99), and recent or current use of antibiotics (OR, 3.13; 95% CI, 1.45-6.77). CONCLUSIONS: Malignancy and neurologic disease are well-recognized conditions that identify patients with lower respiratory tract infections who have a high risk of death within 30 days. An atypical presentation with confusion, lethargy, poor eating, or recent or current antibiotic use also identifies patients, with a high risk of 30-day mortality. PMID- 9342996 TI - Functional disability before myocardial infarction in the elderly as a determinant of infarction severity and postinfarction mortality. AB - BACKGROUND: Functional disability is a common condition among elderly patients. However, to our knowledge, its effect on outcome of myocardial infarction (MI) has not been assessed. Our objectives were to determine whether disability in the activities of daily living measured before MI is a predictor of MI severity and mortality. METHODS: Disability in activities of daily living was measured prospectively in a cohort of 222 patients who were hospitalized with acute MI. Outcome measures were severity characteristics on admission to the hospital (higher Killip class, presence of new Q waves in the first electrocardiogram, and lower systolic blood pressure), and 6-month mortality. RESULTS: Patients with disability before hospitalization were older and had more comorbidity. After adjusting for these factors and for delay in hospital arrival, disability was still significantly associated with clinical severity on admission to the hospital and with mortality (adjusted relative risk of death for patients with disability vs patients without disability, 2.01; 95% confidence interval, 1.23 3.28). Clinical severity and hospital treatment explained the higher mortality of patients with disability. When these factors were added to the previous model, the relative risk of mortality for patients with disability vs patients without disability was 1.24, and the 95% confidence interval was 0.73 to 2.12. CONCLUSIONS: Functional disability in activities of daily living before MI is an important predictor of clinical severity and mortality in elderly patients with MI. This effect is only minimally explained by the older age and higher comorbidity of patients with disability. However, higher clinical severity and lower use of treatment interventions are major determinants of their higher mortality compared with patients without disability. PMID- 9342997 TI - Adverse events after discontinuing medications in elderly outpatients. AB - BACKGROUND: Discontinuation of drug therapy is an important intervention in elderly outpatients receiving multiple medications, but it may be associated with adverse drug withdrawal events (ADWEs). OBJECTIVE: To determine the frequency, types, timing, severity, and factors associated with ADWEs after discontinuing medications in elderly outpatients. PATIENTS: One hundred twenty-four ambulatory elderly participants in 1-year health service intervention trial at the Durham Veterans Affairs General Medicine Clinic in Durham, NC, who stopped taking medications. METHODS: A geriatrician retrospectively reviewed computerized medication records and clinical charts to determine medications no longer being taken and adverse events in the subsequent 4-month period. Possible ADWEs, determined by using the Naranjo causality algorithm, were categorized by therapeutic class, organ system, and severity. RESULTS: Of 238 drugs stopped, 62 (26%) resulted in 72 ADWEs among 38 patients. Cardiovascular (42%) and central nervous system (18%) drug classes were most frequently associated with ADWEs. The ADWEs most commonly involved the circulatory (51%) and central nervous (13%) systems, and 88% were attributed to exacerbations of underlying disease. Twenty six ADWEs (36%) resulted in hospitalization or an emergency department or urgent care clinic visit. Only the number of medications stopped was associated with ADWE occurrence (adjusted odds ratio, 1.89; 95% confidence interval, 1.33-2.67). CONCLUSIONS: Most medications can be stopped in elderly outpatients without an ADWE occurrence. However, when ADWEs occur they resulted in substantial health care utilization. Practitioners should strive to discontinue drug therapy in the elderly but be vigilant for disease recurrence. PMID- 9342998 TI - Stress management and exercise training in cardiac patients with myocardial ischemia. Effects on prognosis and evaluation of mechanisms. AB - BACKGROUND: Previous studies have demonstrated that myocardial ischemia can be elicited by mental stress in the laboratory and during daily life and that ischemia induced by mental stress is associated with an increased risk for future cardiac events in patients with coronary artery disease. OBJECTIVES: To examine the extent to which ischemia induced by mental stress can be modified by exercise stress management, and to evaluate the impact of these interventions on clinical outcomes. METHODS: One hundred seven patients with coronary artery disease and ischemia documented during mental stress testing or ambulatory electrocardiographic monitoring were randomly assigned to a 4-month program of exercise or stress management training. Patients living at a distance from the facility formed a nonrandom, usual care comparison group. Myocardial ischemia was reassessed following treatment, and patients were contacted annually for as long as 5 years to document cardiac events, including death, nonfatal myocardial infarction, and cardiac revascularization procedures. RESULTS: Twenty-two patients (21%) experienced at least 1 cardiac event during a mean (+/- SD) follow up period of 38 +/- 17 months. Stress management was associated with a relative risk of 0.26 compared with controls. The relative risk for the exercise group also was lower than that of controls, but the effect did not reach statistical significance. Stress management also was associated with reduced ischemia induced by mental stress and ambulatory ischemia. CONCLUSION: These data suggest that behavioral interventions offer additional benefit over and above usual medical care in cardiac patients with evidence of myocardial ischemia. PMID- 9342999 TI - Recurrence of venous thromboembolism in patients with familial thrombophilia. AB - BACKGROUND: Treatment of patients with deep vein thrombosis and an antithrombin or protein C or S deficiency is based on case reports and personal experience. OBJECTIVE: To systematically assess the risk for recurrence of venous thromboembolism after a first episode in patients with these deficiencies, a literature review and retrospective family cohort study were performed. METHODS: For the literature review, the annual incidence of a first recurrent venous thromboembolism was assessed for each deficiency by dividing the number of venous thromboembolic events by the number of years at risk. For the family cohort study, 1- and 5-year cumulative incidences of first recurrence were calculated based on medical histories taken in relatives of consecutive patients in whom venous thromboembolism and a deficiency were diagnosed. RESULTS: For the literature review, the annual incidence of a first recurrent venous thromboembolism in patients with antithrombin or protein S deficiency ranged from 13% to 17% and 14% to 16%, respectively. For the family cohort study, the 1- and 5-year cumulative incidences of recurrent venous thromboembolism were 10% (95% confidence interval, 1%-19%) and 23% (95% confidence interval, 10%-36%), respectively. Warfarin sodium (Coumadin) prophylaxis was associated with 2 venous thromboembolic events in 141 years at risk (1.4% per year), in contrast with 19 events in 709 years at risk (2.7% per year) without prophylaxis (difference, 1.3%; 95% confidence interval, -3.5% to 1.0%). CONCLUSIONS: The annual incidence of recurrent venous thromboembolism is high during the first years following a first episode, but seems to decline thereafter. Therefore, our results challenge current practice of prescribing lifelong warfarin therapy after a first or second episode of venous thromboembolism in patients with antithrombin or protein C or S deficiency. PMID- 9343000 TI - Impact of age on the severity, course, and complications of alcohol withdrawal. AB - BACKGROUND: Early identification of alcohol-dependent patients at increased risk for severe or complicated alcohol withdrawal would improve triage and treatment. However, the role of age in predicting alcohol withdrawal outcomes has not been well studied. OBJECTIVE: To assess the impact of age on the severity, course, and complications of alcohol withdrawal. METHODS: We performed a retrospective cohort study of 284 inpatients admitted for alcohol withdrawal between September 1992 and August 1994. Outcomes included alcohol withdrawal severity measured by the revised Clinical Institute Withdrawal Assessment for Alcohol scale, quantity and duration of benzodiazepine therapy, and complications during withdrawal. RESULTS: Initial and maximal withdrawal severity scores, amount of benzodiazepine administered, and duration of benzodiazepine treatment for elevated withdrawal severity scores did not change significantly with age. However, patients aged 60 years and older had increased risk for delirium (adjusted odds ratio [OR], 4.7; 95% confidence interval [CI], 1.5-15.0; P = .008), falls (OR, 3.1; 95% CI, 0.9 11.2; P = .08), and transient dependency in 2 or more activities of daily living (OR, 5.8; 95% CI, 2.9-11.7; P < .001). As age increased, there were significant increases in length of stay (P < .001) and frequency of discharge to an extended care facility (P < .001). CONCLUSIONS: Although alcohol withdrawal severity scores and benzodiazepine requirements were similar across age groups, patients aged 60 years and older were at increased risk for cognitive and functional impairment during withdrawal. These findings support recommendations that older patients with alcohol withdrawal are best treated in closely supervised settings. PMID- 9343001 TI - Quality of care for elderly patients hospitalized with heart failure. AB - BACKGROUND: There is increasing interest in the development of explicit criteria to evaluate the quality of care for patients with heart failure. However, despite this interest, there is a paucity of information about the care of these patients in actual clinical practice across diverse sites. METHODS: We conducted a retrospective medical record review across 9 acute care hospitals in Connecticut. We selected 200 random admissions from each hospital with a principal discharge diagnosis of heart failure in 1994. Hospitals with fewer than 200 cases had 100% of cases selected. Patients with heart failure secondary to severe aortic stenosis, mitral stenosis, or medical illness were excluded. We evaluated the percentage of patients receiving appropriate treatments and interventions as defined by quality-of-care indicators derived from the Agency for Health Care Policy and Research Clinical Practice Guidelines. RESULTS: Data were abstracted from 1623 hospitalizations and the presence of heart failure was validated by chart review in 1535 (95%). In cohorts of ideal candidates for specific interventions, 832 (75%) of 1110 had a left ventricular ejection fraction documented or measured; 346 (86%) of 401 received angiotensin-converting enzyme inhibitors; 38 (14%) of 271 received target doses of angiotensin-converting enzyme inhibitors; 1359 (97%) of 1400 had counseling about medications documented; 90 (6%) of 1400 had counseling about weight documented; 980 (70%) of 1400 had counseling about diet documented; 856 (61%) of 1400 had counseling about exercise and activity documented; and 14 (11%) of 128 smokers had counseling about cessation documented. CONCLUSIONS: These data demonstrate that the documentation of left ventricular systolic function and counseling for diet, weight, activity, and smoking may provide the best opportunities to improve the hospital care of elderly patients with heart failure. The use of angiotensin converting enzyme inhibitors in appropriate patients is relatively high, indicating successful translation of trial results into clinical practice at these hospitals. PMID- 9343002 TI - Risk factors for all-cause and coronary heart disease mortality in the oldest old. The Adventist Health Study. AB - BACKGROUND: The oldest-old population (> or = 84 years of age) is growing rapidly and consumes a disproportionate amount of health care dollars. Risk factors for disease have not been extensively studied in this group. METHODS: A cohort study of non-Hispanic white Seventh-Day Adventists from California allowed follow-up for mortality from 1976 through 1988. Associations between traditional risk factors, consumption of selected foods, and both coronary heart disease (CHD) and all-cause mortality were evaluated in the oldest-old portion of this population, using proportional hazards regression analyses. RESULTS: We observed 364 cases of CHD and 1387 total deaths during 11,828 person-years of follow-up. Men had higher risk of both all-cause mortality and mortality from CHD. The relative risks (RRs) associated with diabetes mellitus were 1.51 (95% confidence interval [CI], 1.24 1.84; P < .001) for all deaths and 1.95 (95% CI, 1.38-2.76; P < .001) for mortality from CHD. The apparent effects of hypertension were small unless subjects were currently taking antihypertensive medications. Compared with those with no regular vigorous activity, subjects who exercised at least 3 times each week had RRs of death of 0.80 (95% CI, 0.70-0.91; P < .001) and 0.74 (95% CI, 0.56-0.97; P < .05) for mortality from CHD. Subjects who consumed nuts 5 times per week had RRs of death of 0.82 (95% CI, 0.70-0.96; P < .01) and 0.61 (95% CI, 0.45-0.83; P < .001) for death from CHD compared with those consuming nuts less than weekly. In men, regular consumption of donuts appeared hazardous for both all-cause mortality (RR, 1.40; 95% CI, 1.05-1.88) and mortality from CHD (RR, 2.10; 95% CI, 1.15-3.81), and consumption of beef 4 times weekly was associated with a 2-fold RR for CHD compared with vegetarians, but there was no increase in risk for women. CONCLUSIONS: Even in the oldest-old, certain traditional risk factors and dietary habits are associated with mortality. PMID- 9343003 TI - Price of adaptation--allostatic load and its health consequences. MacArthur studies of successful aging. AB - BACKGROUND: Exponential growth in the population of older adults presents clinicians with special concerns about factors affecting risks for declines in cognitive and physical functioning. OBJECTIVES: To examine the hypothesis that risks for such declines and for disease outcomes, such as cardiovascular disease, are related to differences in allostatic load, the cumulative physiologic toll exacted on the body over time by efforts to adapt to life experiences. To present an operational definition of allostatic load, along with preliminary evidence of its predictive validity in relation to salient outcomes of aging. METHODS: Data from a longitudinal, community-based study of successful aging were used to develop a measure of allostatic load based on 10 parameters reflecting levels of physiologic activity across a range of important regulatory systems. Allostatic load is the sum of the number of parameters for which the subject was rated in the highest-risk quartile. RESULTS: Higher allostatic load scores were associated with poorer cognitive and physical functioning and predicted larger decrements in cognitive and physical functioning as well as being associated with an increased risk for the incidence of cardiovascular disease, independent of sociodemographic and health status risk factors. CONCLUSIONS: Findings are consistent with the conceptualization of allostatic load as an index of wear and tear on the body, with elevations in allostatic load predicting an increased risk for a decline in cognitive and physical functioning as well as cardiovascular disease in a cohort of older men and women. From a clinical perspective, the concept of allostatic load may provide the basis for a more comprehensive assessment of major risks in the aging process. PMID- 9343004 TI - Physicians who report health insurance fraud and their practice type: health maintenance organization vs fee-for-service. PMID- 9343005 TI - Informing children of their human immunodeficiency virus infection. PMID- 9343006 TI - Patterns of disclosure and perceptions of the human immunodeficiency virus in infected elementary school-age children. AB - OBJECTIVE: To investigate the patterns of disclosure and perceptions of human immunodeficiency virus (HIV) status in a group of HIV-infected elementary school age children. DESIGN: A survey. SETTING: A referred care university hospital center. PATIENTS: All HIV-infected children born before August 31, 1985, and scheduled for ambulatory follow-up between 1984 and 1993 were eligible for the study. A total of 35 HIV-infected (21 asymptomatic and 14 symptomatic) elementary school-age children (aged 5-10 years) were examined between 1990 and 1993. MAIN OUTCOME MEASURES: Semistructured qualitative interviews were used, 1 with the children and 1 with their parents or caregivers. In addition, 3 drawings per child were also analyzed. RESULTS: Partial disclosure was observed in 14 (40%) of the children, and full disclosure of the diagnosis of acquired immunodeficiency syndrome was given to 6 (17%) of the children. Secrecy regarding serostatus was the strategy used by 15 (43%) of the parents or caregivers involving either complete nondisclosure (n = 8) or deception by means of attributing the symptoms to another condition, medical or other (n = 7). Perceived health status and clinical status differed for 11 (31%) of the children. Eight children did not identify any illness causality, and most of the others gave prelogical or concrete-logical explanations. Few children were aware of their parent's infection or disease. CONCLUSION: Human immunodeficiency virus-infected elementary school-age children were exposed to various disclosure patterns regarding their HIV infection or disease, and most children (26/35 [74%]) reported stressful experiences due to HIV regardless of the disclosure patterns. PMID- 9343007 TI - Iron deficiency in 1- to 3-year-old children. A pediatric failure? AB - OBJECTIVE: To determine the prevalence of iron deficiency and iron deficiency anemia in children aged 1 to 3 years in an urban population. DESIGN: Venous blood was measured for levels of hemoglobin, ferritin, free erythrocyte protoporphyrin, and lead in children seen for well-child visits. Children with histories of chronic illness, prematurity, blood dyscrasias, and acute illness were excluded. SETTING: The private practice offices of 4 pediatricians in the New York City area. PATIENTS: A consecutive sample of 504 children aged 1 to 3 years was included. RESULTS: More than one third (35%) of the children demonstrated evidence of iron insufficiency; 7% were iron deficient without anemia, and 10% had iron deficiency anemia. CONCLUSION: Because the association of iron deficiency anemia with mental and psychomotor impairment during the first 2 years of life no longer seems to be in doubt, the high prevalence of iron deficiency anemia found in the 1- to 2-year-old children in this study is disturbing. This suggests the need for greater efforts at the prevention of iron deficiency during the second year of life. PMID- 9343008 TI - Prevalence and features of joint hypermobility among adolescent athletes. AB - OBJECTIVE: To determine the prevalence of joint hypermobility in a group of adolescent, interscholastic athletes. DESIGN: Cross-sectional; descriptive or observational. SETTING: Free preparticipation physical examinations for sports. SUBJECTS: Two hundred and sixty-four athletes (150 male, 114 female; average age, 15.5 years) comprised the entire set of athletes who came to our clinic for free physical examinations. INTERVENTION AND MAIN OUTCOME MEASURES: We screened 264 athletes using the widely accepted Carter-Wilkinson-Beighton method, which examines range of motion at the knees, trunk, fingers, thumbs, and elbows bilaterally and employs a 0 to 9 scoring scheme (5 = hypermobile). We also used an "injury allowance," whereby if an athlete screened positive for only one side of a bilateral test but had a history of injury to the corresponding side, he or she was given an injury allowance point. RESULTS: Thirty-two scored 5 or higher, with another 2 screening positive for hypermobility by the injury allowance, for a total of 34 hypermobile athletes (12.9%). There was a highly significant difference between sexes (P < .001), with 25 female (22%) and 9 male subjects (6%) testing positive. CONCLUSIONS: The overall prevalence of hypermobility and the significant sex difference found in this group of adolescent athletes were similar to nonathletes populations of comparable age. Research on nonathletes has been relied on by many to recommend that hypermobile persons avoid strenuous physical activity; however, research on athletes is less than conclusive. Given that a significant segment of young athletes, especially females, may be hypermobile, prospective studies are warranted to investigate this question before we can justify depriving hypermobile youths of the many known benefits of regular or strenuous exercise. PMID- 9343009 TI - Outcome after exploratory laparoscopy for unexplained abdominal pain in childhood. AB - BACKGROUND: Abdominal pain in childhood is common yet frustrating when unexplained. OBJECTIVE: To describe the clinical features and outcome of 8 children (6 girls and 2 boys; mean[+/- SD] age, 13 +/- 2 years) with unexplained abdominal pain who underwent exploratory laparoscopy. SETTING: All 8 patients were examined at an academic pediatric gastroenterology center and referred for exploratory laparoscopy because of unexplained abdominal pain. Laparoscopy was offered after family agreement to pursue behavioral management if the pain and disability did not improve. RESULTS: In all 8 children, laparoscopy detected an anomaly at a site corresponding to that of the abdominal pain. Findings were adhesions in 7 children (3 colonic, 2 ileocecal, 1 gastric, and 1 appendiceal) and ovarian torsion in 1 child. At a mean follow-up of 12.6 months, the abdominal pain had completely resolved in 6 children, notably improved in 1 child, and continued unchanged in 1 child. Disability completely resolved in 2 of 3 children. CONCLUSIONS: In children with unexplained abdominal pain that is acute in onset, well described, and suggestive of peritoneal involvement, exploratory laparoscopy (1) successfully ends the cycle of abdominal pain in most cases; and (2) commonly identifies abnormalities, usually adhesions. However, whether laparoscopy, the placebo effect, or both promote the healing process is unclear. Further study is needed to develop criteria for referral for laparoscopic evaluation of unexplained abdominal pain. PMID- 9343010 TI - Effect of 2 urban emergency department immunization programs on childhood immunization rates. AB - BACKGROUND: Emergency departments (EDs) are recommended as sites for immunizing children. However, there is little information about the effect of ED immunization programs on immunization rates. OBJECTIVES: To assess the ability of 2 ED immunization programs to vaccinate children and to measure the effect of the programs on immunization rates after the ED visit and 6 months later. DESIGN: A prospective cohort study. Emergency department patients were screened for immunization status, and vaccinations were offered to patients who either were documented to be eligible or were eligible by age and had no documented records. A systematic, sequential sample of those accepting vaccinations (study patients) was compared with a systematic, sequential sample of those not vaccinated (control subjects). Telephone interviews and medical record reviews were performed 6 months after the ED visit to verify dates of immunizations. Results were weighted to reflect the sampling frames of patients screened by the 2 programs. SETTING: Two EDs in New York City (in Manhattan and the Bronx) and the surrounding primary care offices. PATIENTS: Children (aged 0-6 years) screened for immunization status by the ED immunization program during a 10-week period; these included 210 children from the Manhattan ED (106 vaccinated in the ED) and 274 children from the Bronx ED (129 vaccinated in the ED). INTERVENTION: Emergency department immunizations. MAIN OUTCOME MEASURES: Proportion of patients (vaccinated, not vaccinated, and ED population) up-to-date for immunizations (1) at the time of the ED visit, (2) 1 day later, and (3) 6 months later. RESULTS: Two thirds of the patients in each ED had Medicaid, and one tenth were uninsured. At the time of the ED visit, 20% of the vaccinated children in each ED were actually up-to-date and were unnecessarily vaccinated; 74% (Manhattan ED) and 72% (Bronx ED) of the not vaccinated children were up-to-date (the remainder were later determined to have been eligible for vaccinations). One day after the ED visit, and 6 months later, the immunization rates of the vaccinated and not vaccinated children were similar. The results of the weighted analysis were as follows: for the entire ED population screened for immunization status, compared with up-to-date rates at the time of the ED visit, rates 1 day later were 11% (Manhattan ED) and 8% (Bronx ED) higher in each ED (P < .05); and rates 6 months later were the same in the Manhattan ED and 10% lower in the Bronx ED (P < .01). Eighteen percent of all children screened for immunization status were vaccinated; 10 to 15 children were screened and 2 to 4 children were vaccinated per 8-hour ED shift. CONCLUSIONS: This ED immunization program temporarily improved the immunization rates of the ED population, but substantial personnel time was required to achieve these small gains. Urban ED immunization programs are unlikely to be cost-effective. PMID- 9343011 TI - Shared management of children with cancer. AB - OBJECTIVE: To determine the risks and benefits of university-based pediatric oncologists and community-based primary care physicians sharing the management of children with cancer. DESIGN: Physicians participating in shared management of children with cancer were surveyed, and the outcomes of the children were measured. SETTING AND PARTICIPANTS: One hundred thirty-seven community-based primary care physicians participated in the management of the 226 children with cancer in Iowa and western Illinois during the past 15 years. The survival of the 226 children was compared with that of 240 randomly selected children treated using the identical treatment protocols but treated only by pediatric oncologists. INTERVENTION: A 7-point Likert scale questionnaire was completed by 97 (71%) of the participating primary care physicians. RESULTS AND OUTCOME MEASURES: There were no differences in the survival of children using shared management compared with those treated only by pediatric oncologists. Primary care physicians believed that shared management is of economic and psychosocial benefit to patients, improves the treatment choices available to patients, does not require excessive time, and does not result in loss of practice income. The system strengthens the primary care physicians' relationships with oncologists and results in additional referrals to the university-based pediatric oncologists. It is of educational value, is personally satisfying, and provides relief from the stress associated with caring for these families. Primary care physicians would like to see this system expanded to include other children with special health care needs. CONCLUSION: The shared-management approach to care is a viable attractive option of health care provision for children. PMID- 9343012 TI - Herpes simplex virus-associated erythema multiforme in prepubertal children. AB - OBJECTIVE: To examine clinical associations, evolution of the condition, and response to treatment of erythema multiforme (EM) in prepubertal children. DESIGN: A retrospective case series evaluation of children younger than 13 years with EM. SETTING: Ambulatory care university hospital. PATIENTS: Referral patients from pediatricians serving a population of 3.2 million. INTERVENTIONS: Results of treatment of each EM episode with topical acyclovir or oral acyclovir at a dose of 25 mg/kg per day and 6-month prophylaxis with oral acyclovir at a dose of 20 mg/kg per day were evaluated. OUTCOMES: Age at EM onset, preceding illness, and number and duration of episodes during a 3-year period were recorded. RESULTS: Twelve children (7 boys and 5 girls) in whom herpes simplex virus (HSV)-associated EM developed were evaluated. Preceding lesions were herpes labialis in 8 children and herpes facialis in 2 children. Two children had no obvious HSV lesion. The mean age at onset of disease was 8.1 years, and the mean time from the preceding HSV to the onset of skin lesions was 3.9 days (range, 0 11 days). Episodes of EM lasted a mean of 10.6 days. In 9 children, the EM was recurrent, with a mean of 2.6 episodes per year. All 12 children, including those with negative viral cultures for HSV or no HSV history had HSV detected in their target lesions by polymerase chain reaction amplification of DNA obtained from skin biopsy specimens. Six of 12 children were treated with oral acyclovir at a dose of 25 mg/kg per day for 1 or more individual episodes, without reduction in the episode. Three children underwent 6-month prophylaxis with oral acyclovir at a dose of 20 mg/kg per day and remained disease free during treatment. After discontinuation of the prophylactic treatment with acyclovir, 1 child relapsed at 4 months. The other 2 children had no further episodes during a 3-year period. CONCLUSIONS: The HSV-associated EM is a recurrent disease that can be precipitated by sun exposure and does not progress to Stevens-Johnson syndrome. Childhood HSV-associated EM may be unresponsive to treatment with oral steroids or oral or topical acyclovir. Frequent recurrences of EM may be abrogated by prophylactic treatment with acyclovir. PMID- 9343013 TI - Psychosocial outcome of children evaluated for short stature. AB - OBJECTIVE: To assess the psychosocial functioning of adults who were evaluated as children for short stature and were not treated with human growth hormone. DESIGN: Inception cohort study. SETTING: Hospital-based pediatric endocrinology clinic. PARTICIPANTS: From 1975 to 1980, medical record review indicated that 181 of the children referred to our clinic for concerns about short stature were non growth hormone deficient. In 1992 and 1993, we were able to recruit 35 of these patients for a follow-up study. Eligible subjects were at least 18 years of age at the time of follow-up. MAIN OUTCOME MEASURES: Standardized self-report questionnaires assessed various domains of psychosocial adjustment. Also, a brief test of intellectual functioning was administered and subjects underwent a semistructured in-person interview to evaluate pragmatic functioning and experiences associated with short stature. RESULTS: Few significant differences between the study sample and standardization samples were found on measures of psychosocial and intellectual functioning. Within-group childhood height during the first evaluation appointment was not significantly associated with most adult measures of psychosocial adjustment. Shorter adult stature was significantly associated with lower educational achievement, lower self-esteem, and greater emotional distress. CONCLUSIONS: The absence of significant psychosocial distress or impairment in these subjects brings into question one basis for hormonal treatment for non-growth hormone deficient short stature; that short stature in childhood is likely to lead to psychological dysfunction in adulthood. The results, however, also suggest that shorter stature in adulthood may constitute a psychosocial stressor, increasing vulnerability across several domains. PMID- 9343014 TI - Spontaneous eye blinking, a measure of dopaminergic function, in children with acquired immunodeficiency syndrome. AB - OBJECTIVE: To investigate possible alterations in dopaminergic function in children with acquired immunodeficiency syndrome by evaluating spontaneous eye blink rate, a putative measure of central dopaminergic function. DESIGN: Evaluation of previously videotaped test sessions of a consecutive case series of 50 children (mean age, 5.2 years; range, 2-12 years) with acquired immunodeficiency syndrome. SETTING: Government medical research center. RESULTS: Intrarater reliability was high, expected co-variation of blink rate with age and concurrent mental activity were confirmed, and obtained rates were similar to published data. Higher blink rates, suggestive of increased dopaminergic function, were associated with more severe cortical atrophy (P < .05) and white matter abnormality (P < .05) on computed tomographic brain scans. The presence or severity of basal ganglia calcifications did not seem to influence blink rate. In addition, higher blink rates were associated with higher ratings of depressed affect (P < .05) and lower ratings of hyperactive behaviors (P < .05) during other test activities. CONCLUSIONS: The higher blink rates in human immunodeficiency virus-infected children with more severe cortical abnormalities suggest increased central dopamine activity compared with that in children without cortical computed tomographic brain scan abnormalities. Thus, as a result of structural brain abnormalities, neurotransmitter levels in children with acquired immunodeficiency syndrome may vary and this may be reflected in their socioemotional functioning. PMID- 9343015 TI - Childhood lead poisoning and vinyl miniblind exposure. AB - OBJECTIVE: To determine the contribution of vinyl miniblinds to childhood lead poisoning. DESIGN: A descriptive investigation was undertaken to estimate attributable risk among all reported childhood lead poisoning cases in North Carolina for which home environmental sampling was conducted between March and August 1996. PARTICIPANTS: Ninety-two children, aged 6 to 72 months, identified through a statewide screening program were included. Blood lead and environmental sampling test results were obtained from routine surveillance data collected for all lead-poisoned children. RESULTS: Exposure to vinyl miniblinds with dust lead levels of 100 micrograms/ft2 or more occurred for 44 (48%) of the lead-poisoned children; 25 (27%) of the children were exposed to levels of 500 micrograms/ft2 or more. Vinyl miniblinds were the predominant source (ie, other major sources of lead were not identified) for 8 (9%) of the children. Overall, the median dust lead level for vinyl miniblind field samples was 590 micrograms/ft2, and the highest level reported was 73,302 micrograms/ft2. Even new vinyl miniblinds manufactured before July 1996 contained dust lead levels that on average exceeded 100 micrograms/ft2. The levels for recently available nonleaded vinyl miniblinds were below the limits of detection. CONCLUSIONS: Vinyl miniblinds, introduced into this country 10 years ago, with sales estimated at 30 million sets a year, include brands containing lead. Although new formulations with no lead added are available, millions of children may still be at risk because a product recall has not been issued (ie, lead-contaminated vinyl miniblinds are still in general use). In addition, the risk assessment evaluations proposed in lieu of universal blood lead screening for low-risk communities could overlook children with exposure to this source. PMID- 9343017 TI - Sex assignment in the neonate with intersex or inadequate genitalia. PMID- 9343016 TI - Pediatric injury prevention. Preparing residents for patient counseling. AB - OBJECTIVES: To describe counseling practices on injury prevention and barriers to patient counseling by pediatric residents and determine whether education about injury prevention or use of educational aids promotes this activity in resident ambulatory practices. DESIGN: Cross-sectional mail survey. SETTING: Accredited US pediatric residency programs. PARTICIPANTS: Pediatric chief residents. MAIN OUTCOME MEASURE: Reported frequencies of patient counseling performed on various injury prevention topics. RESULTS: All residents reported that they were expected to educate patients and families about injury prevention in the continuity clinic setting. Almost all residents (98.5%) reported that they counseled on at least 1 injury prevention topic. On all topics except for poisoning prevention, residents were most likely to counsel patients and families on those topics about which they had received education. Additionally, those residents familiar with the American Academy of Pediatrics The Injury Prevention Program included more injury prevention topics in their counseling repertoire (P = .01). The most frequently identified barriers to counseling included lack of information about the topic and lack of time in the visit. CONCLUSIONS: Most pediatric residents counsel their patients and families on a variety of injury prevention topics. This activity is promoted by the education offered during residency training. Focused efforts should be made to educate residents about those injury topics not being taught and to address counseling barriers with educational interventions that promote prevention counseling during patient visits. PMID- 9343018 TI - Management of intersexuality. Guidelines for dealing with persons with ambiguous genitalia. AB - Following the publication of our article about a classic case of sex reassignment, the media attention was rapid and widespread, as was the reaction of many clinicians. Some wanted to comment or ask questions, but many contacted us directly or indirectly, asking for specific guidelines on how to manage cases of traumatized or ambiguous genitalia. PMID- 9343019 TI - Radiological case of the month. Varicella mimicking pulmonary metastasis in a patient with rhabdomyosarcoma. PMID- 9343020 TI - Radiological case of the month. Congenital cystic adenomatoid malformation of the lung and pulmonary blastoma. PMID- 9343021 TI - Picture of the month. Chilblains (pernio). PMID- 9343022 TI - Pathological case of the month. Retinoblastoma with invasion of the optic nerve. PMID- 9343023 TI - Pathological case of the month. Bronchopulmonary sequestration, intralobar type. PMID- 9343024 TI - Sex reassignment at birth: long-term review and clinical implications. PMID- 9343025 TI - Sex reassignment at birth. PMID- 9343026 TI - Sex reassignment at birth. PMID- 9343027 TI - Serotonin syndrome after sertraline overdose in a 5-year-old girl. PMID- 9343028 TI - Bicycle-riding circumstances and injuries in school-age children. PMID- 9343029 TI - Anticoagulation in chronic nonvalvular atrial fibrillation: a critical appraisal and meta-analysis. AB - OBJECTIVE: To assess the outcomes associated with warfarin treatment of patients with chronic nonvalvular atrial fibrillation (CNVAF) for prevention of primary stroke. DATA SOURCES: MEDLINE was searched for literature published from 1987 to August 1996. Search terms used were 'atrial fibrillation' and 'anticoagulants'. STUDY SELECTION: Five published randomized controlled trials concerning primary stroke prevention. DATA EXTRACTION: Data were pooled across trials to estimate the magnitude of the effect for each of nine reported end-points. The annual probability of occurrence of each outcome was calculated, including standard errors and Mantel-Haenszel significance tests with 95% CIs. DATA SYNTHESIS: In view of the lack of blinded assessment and documented low inter-rater reliability of soft neurological end-points, the analysis was limited to the relatively objective end-points of major strokes, fatal strokes, major bleeding and fatal bleeding. Warfarin did not reduce the incidence of fatal strokes to a statistically significant extent, nor was incidence of fatal bleeding increased significantly. Warfarin reduced the absolute annual incidence of major strokes in patients with CNVAF by 0.89%, while at the same time it increased the absolute annual risk of major bleeding incidents by 1.8%. Though small, these differences were statistically significant. CONCLUSIONS: On balance, the margin between expected benefit and harm for warfarin prophylaxis in patients with CNVAF is uncomfortably thin. These results and conclusions differ from those of a previously published meta-analysis of these same studies. PMID- 9343030 TI - Inhaled nitric oxide reduction in systolic pulmonary artery pressure is less in patients with decreased left ventricular ejection fraction. AB - OBJECTIVE: To assess whether inhaled nitric oxide decreases pulmonary artery pressure in patients with depressed left ventricular ejection fraction. DESIGN: Randomized, blinded, crossover clinical trial. SETTING: Tertiary care university referral hospital. PATIENTS: Thirty-three patients with pulmonary hypertension and left ventricular dysfunction or valvular heart disease were recruited by convenience. INTERVENTIONS: Systolic pulmonary artery pressure was measured by Doppler echocardiography during randomized inhalation of either 20 ppm or 40 ppm nitric oxide in 30% oxygen as well as during control periods without nitric oxide. MAIN RESULTS: Systolic pulmonary artery pressure was significantly (P < 0.05) decreased with 20 ppm nitric oxide (53.4 +/- 13.9 mmHg) and 40 ppm nitric oxide (53.1 +/- 14.4 mmHg) compared with either initial control (55.8 +/- 15.3 mmHg) or terminal control (56.3 +/- 15.2 mmHg) values. The regression equation for the change in systolic pulmonary artery pressure (y) as predicted by the left ventricular ejection fraction (x) alone for 20 ppm nitric oxide was y = 13.8x 2.9; R2adj = 0.30, P < 0.0001. For 40 ppm nitric oxide alone, the regression equation was y = 16.3x-3.3; R2adj = 0.25, P < 0.0001. Left ventricular ejection fraction was the most explanatory independent variable in the multivariate equation for nitric oxide-induced change in systolic pulmonary artery pressure (R2 = 0.61, P = 0.0000). The change in systolic pulmonary artery pressure was 5.1 +/- 5.2 versus 0.8 +/- 4.9 mmHg (P < 0.0000) in patients with left ventricular ejection fractions greater than 0.25, and 0.25 or less, respectively. CONCLUSIONS: These data imply that in patients with left ventricular ejection fraction of 0.25 or less, nitric oxide may not decrease systolic pulmonary artery pressure. Nitric oxide inhalation may result in a paradoxical increase in systolic pulmonary artery pressure in patients with severely depressed left ventricular ejection fraction. This effect would significantly limit the therapeutic role of nitric oxide in patients with severe heart failure. PMID- 9343031 TI - Clinical outcomes of patients more than one year following randomization in the Canadian Coronary Atherectomy Trial (CCAT). AB - BACKGROUND: The Canadian Coronary Atherectomy Trial (CCAT) assessed, in a randomized comparison, the clinical and angiographic outcomes following atherectomy with those following balloon angioplasty for the treatment of de novo lesions in the proximal one-third of the left anterior descending artery (LAD). Although the procedural success rate was somewhat higher and the postprocedure lumen larger in patients treated with atherectomy, lumen dimensions, restenosis rates and clinical outcomes were similar in the two groups at six months. To determine whether late differences emerged between the groups, clinical follow-up was obtained at a median of 18 (range 10 to 31) months after randomization. METHODS AND RESULTS: Patients were contacted monthly by telephone for the first six months. Subsequent follow-up information was obtained in 272 (99%) of the 274 randomized patients via a clinic visit or telephone interview with the patient and/or a relative. Additional information was obtained from the referring physician as required. There were no differences in adverse events between the two groups during follow-up. In patients randomized to atherectomy compared with balloon angioplasty, death occurred in 1.5% versus 2.2% (cardiac death 0.7% versus 0.7%); myocardial infarction in 5.1% versus 5.9% (Q wave 1.5% versus 1.5%); coronary bypass surgery in 13.1% versus 12.6%; and repeat target lesion intervention in 22.6% versus 21.5%. Persistent or recurrent Canadian Cardiovascular Society class III/IV angina not treated by a further intervention was present in 1.5% versus 2.2%. The combined end-point of death or nonfatal myocardial infarction occurred in nine (6.6%) versus 11 (8.1%) patients and any adverse cardiac event in 50 (36.5%) versus 53 (39.3%). Multivariate logistic regression indicated that unstable angina, reference vessel size and preprocedure minimum lumen diameter were the only variables independently associated with adverse events. CONCLUSIONS: The initial choice of directional atherectomy or balloon angioplasty had no impact on clinical outcome over a period of 18 months in this patient population. With either technique, just over 60% of patients with proximal LAD disease experienced sustained symptomatic improvement without an adverse event following a single procedure, and 80% achieved this status following a repeat percutaneous intervention. PMID- 9343032 TI - Antihypertensive effects of perindopril treatment in adult spontaneously hypertensive rats. AB - OBJECTIVE: To determine the effect of perindopril treatment and treatment withdrawal in the prevention of hypertension in adult male spontaneously hypertensive rats (SHR). ANIMALS AND METHODS: Beginning at 15 weeks of age, male SHR were treated with either distilled water (control) or different daily dosages of perindopril (1, 2 or 4 mg/kg) by gavage for 10 weeks, followed by 10 weeks of treatment withdrawal. Systolic blood pressure, heart rate and body weight of adult SHR were determined at regular intervals before, during and after the treatment withdrawal periods. At the end of the treatment withdrawal period, plasma and tissue samples were taken for measurement of noradrenaline levels. Angiotensin-converting enzyme (ACE) activity in the plasma from adult SHR and Wistar-kyoto (WKY) rats treated with perindopril 4 mg/kg for two weeks was measured by a radioassay method 6 and 24 h after treatment. RESULTS: Treatment with perindopril caused a dose-dependent lowering of blood pressure in SHR during the 10-week treatment. After withdrawal of the treatment, persistent lowering of blood pressure was found in SHR treated with higher dosages (2 or 4 mg/kg), but not in the 1 mg/kg group. There was no difference in the tissue level of noradrenaline among the control group and SHR previously treated with perindopril. In SHR and WKY treated with perindopril for two weeks, plasma level of ACE activity was reduced longer than 24 h compared with their respective controls. CONCLUSIONS: Chronic treatment of adult SHR with perindopril has a dose dependent effect on the blood pressure of these animals both during and after withdrawal of treatment, but such a treatment had no long term effects on the noradrenaline levels in various tissues. PMID- 9343033 TI - Oral propafenone for rapid conversion of recent onset atrial fibrillation--a review. AB - OBJECTIVE: To review comparative studies evaluating oral propafenone for restoring sinus rhythm in recent onset atrial fibrillation. DATA SOURCES: A MEDLINE search of the English-language literature (1966 to 1996) along with any referenced articles not identified by MEDLINE. STUDY SELECTION: Because intravenous propafenone is not marketed in Canada, only studies evaluating oral propafenone were included. Studies were selected if they compared oral propafenone with placebo or other antiarrhythmic agents for converting recent onset atrial fibrillation to normal sinus rhythm. DATA SYNTHESIS: Propafenone is often used as a first-line agent for pharmacological cardioversion of atrial fibrillation. In earlier studies, the efficacy of propafenone in restoring sinus rhythm was reported to be low with conversion rates of 6% to 62%. Many of these studies were noncomparative and often included patients with refractory, chronic atrial fibrillation or employed suboptimal doses of propafenone. More recently propafenone has been evaluated in the treatment of recent onset atrial fibrillation by using a single 600 mg oral loading dose. Success rates of 76% at 8 h and 83% at 12 h following the loading dose are reported. The incidence of atrial flutter during active treatment was similar to that with placebo, with the majority exhibiting 2:1 or greater atrioventricular conduction ratios and heart rates 150 beats/min or less. CONCLUSIONS: A single 600 mg oral dose of propafenone is highly effective at restoring sinus rhythm in patients with acute onset atrial fibrillation with few adverse effects. The small studies reviewed cannot lead to definitive conclusions about the safety of propafenone without prior administration of agents for rate control. PMID- 9343034 TI - Mitral balloon valvuloplasty in pregnancy: limiting radiation and procedure time by using transesophageal echocardiography. AB - A 35-year-old female with mitral stenosis was admitted to hospital in her 23rd week of gestation for management of disabling dyspnea and paroxysmal nocturnal dyspnea. She was in New York Heart Association functional class III and underwent successful percutaneous balloon mitral valvuloplasty with significant improvement of her symptoms. To limit radiation exposure and procedure time, transesophageal echocardiography was used in combination with pulsed fluoroscopy. The remainder of her pregnancy was uncomplicated and she gave birth to a healthy baby at 38 weeks' gestation. PMID- 9343035 TI - Unstable angina associated with sertraline. AB - An 81-year-old woman reported with chest pain occurring shortly after initiating treatment with sertraline. She had no prior history of cardiovascular disease. She developed nausea and malaise 4 h after her first dose, which resulted in avoidance of further treatment. After voluntarily reinitiating sertraline 10 days later, she again developed nausea and malaise but persisted with treatment. On the second day, her gastrointestinal symptoms were accompanied by crushing retrosternal chest pain radiating to both arms and resolving spontaneously after 10 mins. Following the third dose of sertraline, the patient experienced severe and persistent crushing retrosternal chest pain radiating to both arms. She was hospitalized with a diagnosis of unstable angina and treated with acetylsalicylic acid, intravenous heparin and nitroglycerin. The temporal relationship of chest pain onset following ingestion of sertraline is strongly suggestive of an adverse medication effect. PMID- 9343036 TI - Ticlopidine-carbamazepine interaction in a coronary stent patient. AB - Neurological symptoms occurred in association with increased carbamazepine levels because of a probable drug interaction between ticlopidine and carbamazepine in an epileptic patient undergoing coronary stenting. Physicians should be aware of this possible drug interaction. PMID- 9343037 TI - Idiopathic dilation of the right atrium: case report and survey of the literature. AB - Idiopathic dilation of the right atrium (IDRA) is a rare cardiac anomaly of unknown etiology. Whether it is an acquired or congenital lesion is controversial. A case of IDRA detected in utero and confirmed postnatally is reported. The postnatal course was complicated by the development of atrial flutter successfully treated with sotalol. A review of the literature concerning IDRA is presented. PMID- 9343049 TI - Smoking cessation interventions. PMID- 9343038 TI - Lupus-related mitral valve disease: embolic coronary occlusion as a unique cause of myocardial infarction. AB - Early diagnosis and greatly improved treatment have markedly altered the clinical evolution of systemic lupus erythematosus; the pattern of cardiac involvement in lupus has also changed. To illustrate this, a young woman who died from severe mitral valve disease, including a coronary embolus from verrucous endocarditis, is presented. Mitral valve involvement in lupus is no longer limited to the small benign lesions described by Libman and Sacks. PMID- 9343050 TI - Panic states and chest pain. PMID- 9343051 TI - Hepatitis B virus infection, hepatitis B vaccine, and hepatitis B immune globulin. AB - Hepatitis B virus (HBV) infection is a major health problem in the United States; in 1995, approximately 128,000 cases occurred. Transmission of HBV occurs primarily by blood exchange (eg, by shared needles during injection drug use) and by sexual contact. Persons infected early in life are much more likely to become chronically infected than those infected during adulthood: as many as 90% of infants infected perinatally develop chronic infection and up to 25% will die of HBV-related chronic liver disease as adults. Clinical signs of acute hepatitis occur in about 50% of infected adults but in only 5% of infected preschool-aged children. In the United States, hepatitis B vaccine is currently made by recombinant DNA technology using baker's yeast. Preexposure vaccination results in protective antibody levels in almost all infants and children (> 95%) and healthy adults younger than 40 years of age (> 90%). The most common adverse event following administration of hepatitis B vaccine is pain at the injection site, which occurs in 13% to 29% of adult and 3% to 9% of children. A comprehensive hepatitis B vaccination policy is now recommended that includes (1) routine infant vaccination; (2) catch-up vaccination of 11- to 12-year-olds who were not previously vaccinated; (3) catch-up vaccination of young children at high risk for infection; (4) vaccination of adolescents and adults based on lifestyle or environmental, medical, and occupational situations that place them at risk; and (5) prevention of perinatal HBV infection. PMID- 9343052 TI - Physician satisfaction reflects changes in health care landscape. PMID- 9343053 TI - Changing nature of physician satisfaction with health maintenance organization and fee-for-service practices. AB - BACKGROUND: Managed care practice arrangements, or health maintenance organizations (HMOs), are sufficiently mature to examine whether physicians' level of satisfaction has changed as managed care has developed. This study compares Dane County, Wisconsin, physicians' satisfaction with HMO and fee-for service (FFS) practices in 1986 with that of 1993 and examines factors that contribute to satisfaction in an HMO-dominated environment. METHODS: Cross sectional surveys were mailed to all Dane County physicians in active practice in 1986 and 1993. Physician overall support for HMO development and satisfaction with work situation was measured with single items. Overall satisfaction and clinical freedom within HMO and FFS practices were measured using statistically reliable scales. RESULTS: Significantly more physicians were supportive of the development of HMOs in 1993 than in 1986, and more than two thirds of physicians in 1993 were satisfied in their current work situation. Primary care physicians were significantly more satisfied than subspecialists across most dimensions of satisfaction. Perceived clinical freedom and satisfaction with income continued to be major predictors of satisfaction in 1993 as in 1986. While physicians' satisfaction with HMO practice remained stable, their satisfaction with FFS practice was significantly lower in 1993 than in 1986. Satisfaction with Medicare practice, which was not measured in 1986, was significantly less than with HMO or FFS practice in 1993. CONCLUSIONS: Analyses suggest that primary care physicians are more satisfied than subspecialists with their HMO practice because of their greater satisfaction with HMO-generated income and the expanded clinical freedom they have in HMO practice. An across-the-board decline in satisfaction with FFS practice may be attributable to diminishing clinical freedom resulting from indemnity carriers' increasing micromanagement of patient care. PMID- 9343054 TI - Prevalence of subtypes of low back pain in a defined population. AB - BACKGROUND: For two generations, some back care specialists have emphasized that clinical low back pain is composed of a number of specific syndromes, such as sacroiliitis or trigger points, but the prevalence of these syndromes outside of specialized clinics remains unknown. The purpose of this study was to describe the prevalence of subtypes of low back pain in a defined population. METHODS: The setting was a group model HMO with a population of 54,000. We used a formal group process involving clinicians from many disciplines, long interviews, critical appraisal of the literature, case discussions, and clinical audits to define a set of subtypes of low back pain. Trained physical therapists assessed subtypes in all patients referred for low back pain over a 9-month period, from July 1992 to April 1993. RESULTS: Of the 213 patients evaluated for low back pain, 72% had acute pain (< 3 months) and only 15% had work-related injury. After classification into subtypes, 32% had acute low back strain, 28% had radicular syndromes, 14% had chronic back strain, 10% had sacroiliac syndromes, 6% had posterior facet syndrome, and the remaining 10% included 12 different syndromes. Only about 10% had more than one clinical syndrome. CONCLUSIONS: A limited number of subtypes of low back pain make up the vast majority of low back pain seen in the population at large. Attention to subtypes may provide a way to improve primary care management of low back pain. PMID- 9343055 TI - Variations between family physicians and obstetricians in the evaluation and treatment of preterm labor. AB - BACKGROUND: The purpose of this study was to examine and compare the approaches of family physicians and obstetricians when evaluating and managing women with preterm contractions or labor. METHODS: A survey questionnaire examining physician practice characteristics was sent to a random sample of specialists in obstetrics and family practice in 10 states. Responses were received from 54% of individuals in active practice. RESULTS: When asked their three most common treatment strategies for women with preterm labor, family physicians were more likely than obstetricians to select beta-agonists and hydration. Obstetricians were more likely to include magnesium sulfate and nifedipine in their treatment plans. For women with preterm contractions and no change in cervix, obstetricians were more likely to select either of the two short-term tocolytic therapies, while family physicians were more likely to select less aggressive therapy approaches. When adjusted for the facility in which they practiced and the number of years of experience, family physicians were nearly one half as likely as obstetricians to use tocolytics to treat women who had contractions but no cervical changes. CONCLUSIONS: In general, obstetricians were more likely to select more aggressive therapy for women with premature contractions without changes in cervix. Since it is unlikely that patient preferences would influence the choice of strategies for preterm labor, it is likely that these results reflect true differences in physician practice patterns based on physician specialty. PMID- 9343056 TI - Family physicians' attitudes about and use of clinical practice guidelines. AB - BACKGROUND: The use of clinical guidelines is one strategy intended to improve health care quality, rein in costs, and standardize medical practice. Clinical guideline development has been prodigious, while less effort has been expended on the guidelines' dissemination and implementation. This study examines family physician attitudes toward and perceived uses of clinical guidelines in practice. METHODS: A survey questionnaire was sent to 978 family physicians in Upstate New York to assess their confidence in clinical guidelines developed or endorsed by organizations and the perceived usefulness of such guidelines in practice. Descriptive analyses, chi-square tests, and comparison of means (one-way ANOVA) were conducted. RESULTS: After two mailings, the response rate was 43%. Most respondents perceived clinical guidelines as effective educational tools that should improve the quality of patient care, but were concerned about their potential regulatory intrusion into practice. Solo practitioners expressed more negative attitudes regarding clinical guidelines than physicians in non-solo practices. Respondents had greater confidence in clinical guidelines developed or endorsed by their professional society, the Centers for Disease Control and Prevention, the United States Preventive Services Task Force, and the National Institutes of Health, but less in those by insurance companies or state health departments. The reported adoption rate of clinical guidelines was low. The most preferred methods for adoption were continuing medical education and practice interventions. CONCLUSIONS: Family physicians found clinical guidelines to be valuable educational tools but were divided on their potential regulatory role. If clinical guidelines are to improve quality in practice, they must be more effectively disseminated and implemented. To broaden physicians' adoption of clinical guidelines, further research into dissemination and implementation methods is warranted, along with wider endorsement of guidelines by those whom family physicians trust. PMID- 9343057 TI - Missed opportunities for prevention: smoking cessation counseling and the competing demands of practice. AB - BACKGROUND: Smoking cessation advice is an effective intervention for the control of tobacco use. The objective of this study was to assess and describe the rates of smoking status assessment and smoking cessation advice provided by physicians during ambulatory office visits with respect to physician specialty, type of visit, and number of problems addressed at the visit. METHODS: We used a cross sectional survey of patient visits to the offices of nonfederally employed, office-based physicians participating in the 1992 National Ambulatory Medical Care Survey (n = 1558). RESULTS: Physicians reported knowing the smoking status of their patients in 66% of outpatient visits. The rate of assessment was similar for generalists and specialists. Cardiologists and generalists, except for pediatricians, showed discernible rates of smoking cessation advice (medians ranging from 14% to 50%), whereas obstetrician/gynecologists and other specialists had negligible rates. For tobacco-related visits, generalists and specialists had comparable rates of cessation advice to identified smokers. For non-tobacco-related visits, generalists had higher rates than specialists (22% vs 10%; P < .001). CONCLUSIONS: Although a substantial majority of smokers are reportedly identified by physicians during ambulatory visits, a large number of identified smokers are not receiving smoking cessation counseling. Patients seen by generalists are more likely to receive smoking cessation advice. Physicians appear to prioritize smoking cessation advice based on diagnosis at the time of the visit. PMID- 9343096 TI - Descriptive epidemiology of asthma. PMID- 9343097 TI - The cellular and mediator basis of asthma in relation to natural history. PMID- 9343098 TI - Environmental factors. PMID- 9343099 TI - Towards prevention. PMID- 9343100 TI - Treatment of acute asthma. PMID- 9343101 TI - Limitations of current treatment. PMID- 9343102 TI - The link between low-LET dose-response relations and the underlying kinetics of damage production/repair/misrepair. AB - PURPOSE: To review current opinion on the production and temporal evolution of low-LET radiobiological damage. METHODS: Standard cell survival models which model repair/misrepair kinetics in order to quantify dose-response relations and dose-protraction effects are reviewed and interrelated. Extensions of the models to endpoints other than cell survival, to multiple or compound damage processing pathways, and to stochastic intercellular damage fluctuations are surveyed. Various molecular mechanisms are considered, including double strand breaks restitution and binary misrepair. CONCLUSIONS: (1) Linking dose-response curves to the underlying damage production/processing kinetics allows mechanistic biological interpretations of observed curve parameters. (2) Various damage processing pathways, with different kinetics, occur. (3) Almost every current kinetic model, whether based on binary misrepair or saturable repair, leads at low or intermediate doses to the LQ (linear-quadratic) formalism, including the standard (generalized Lea-Catcheside) dependence on dose protraction. (4) Two track (beta) lethal damage is largely due to dicentric chromosome aberrations, but one-track (alpha) lethal damage is largely caused by other mechanisms such as point mutations in a vital gene, small deletions, residual chromosome breaks, induced apoptosis, etc. (5) A major payoff for 50 years of radiobiological modelling is identifying molecular mechanisms which underly the broadly applicable LQ formalism. PMID- 9343103 TI - A charged-particle microbeam: I. Development of an experimental system for targeting cells individually with counted particles. AB - Charged-particle microbeams provide a unique opportunity to control precisely, the dose to individual cells and the localization of dose within the cell. The Gray Laboratory is now routinely operating a charged-particle microbeam capable of delivering targeted and counted particles to individual cells, at a dose-rate sufficient to permit a number of single-cell assays of radiation damage to be implemented. By this means, it is possible to study a number of important radiobiological processes in ways that cannot be achieved using conventional methods. This report describes the rationale, development and current capabilities of the Gray Laboratory microbeam. PMID- 9343104 TI - A charged-particle microbeam: II. A single-particle micro-collimation and detection system. AB - The use of a charged-particle microbeam provides a unique opportunity to control precisely, the number of particles traversing individual cells and the localization of dose within the cell. The accuracy of 'aiming' and of delivering a precise number of particles crucially depends on the design and implementation of the collimation and detection system. This report describes the methods available for collimating and detecting energetic particles in the context of a radiobiological microbeam. The arrangement developed at the Gray Laboratory uses either a 'V'-groove or a thick-walled glass capillary to achieve 2-5 microns spatial resolution. The particle detection system uses an 18 microns thick transmission scintillator and photomultiplier tube to detect particles with > 99% efficiency. PMID- 9343105 TI - Inactivation of individual mammalian cells by single alpha-particles. AB - PURPOSE: To measure clonogenic death of Chinese hamster V79 cells following exposure to a defined number of 4.3 MeV alpha-particles (track-averaged LET = 105 keV/micron). MATERIALS AND METHODS: Cells were irradiated at the radiobiological facility installed at the TTT-3 Tandem accelerator in Naples by using a 'Biostack' approach, which allows the positions of incident tracks relative to cells to be carefully determined. Subcellular structure was identified by fluorescence microscopy, while tracks were visualized by LR-115 solid state nuclear track detectors. RESULTS: Particle hits in the cytoplasm did not significantly affect cell survival, yet survival probability decreased exponentially as a function of the number of nuclear traversals. Measured probability of surviving to exactly one 4.3 MeV alpha-particle traversal in the cell nucleus was 0.67 +/- 0.10. Inactivation cross-section was substantially higher than expected from conventional survival curves. However, folding of the data with Poisson statistics showed that survival level expected if a mean of one alpha-particle goes through a nucleus is higher than the measured value after exactly one particle traversal. CONCLUSIONS: V79 cells have about 67% probability to survive a single alpha-particle traversal in the cell nucleus. Single-particle survival curves are consistent with conventional dose-survival relationships, once Poisson distribution of traversals is taken into account. PMID- 9343106 TI - Spontaneous premature chromosome condensation and mitotic catastrophe following irradiation of HeLa S3 cells. AB - PURPOSE: To study the mechanisms underlying the loss of G2/M checkpoint control which leads to mitotic catastrophe in human tumour cells following exposure to ionizing radiation. MATERIALS AND METHODS: Asynchronous HeLa S3 cells were irradiated with doses of 5, 10 and 20 Gy X-rays. Cell-cycle progression and cyclin B1 levels were measured using bivariate flow-cytometric techniques as a function of time after irradiation. As indicators of mitotic catastrophe, the appearance of spontaneous premature chromosome condensation (SPCC) and cells presenting nuclear fragmentation were analysed using microscopy. Cyclin B1 dependent kinase activity was determined in immunoprecipitates and analysed using gel electrophoresis. RESULTS: After X-irradiation of HeLa cells, delays in late S and G2 phases of the cell cycle were followed by SPCC and nuclear fragmentation, both indicative of mitotic catastrophe. The kinetics of appearance of cells that had apparently undergone mitotic catastrophe (i.e. the fraction of cells exhibiting nuclear fragmentation) was independent of the dose-dependent radiation induced division delay, while the extent of fragmentation (expressed as the number of nuclear fragments per fragmented cell) did increase with dose. Also observed was a 5-fold elevation of cyclin B1 levels in late S/G2 cells, which correlated temporally with the observed delays late in the cell cycle. Following the appearance of elevated cyclin levels, cyclin B1-associated histone H1 kinase activity showed similar increases; these increases in kinase activity occurred prior to increases in the fraction of cells exhibiting nuclear fragmentation. CONCLUSIONS: In human cells, cyclin B1 gene expression occurs in late S and G2 phases, and thus the increase observed in this protein may be due to the increased time spent by cells in these phases as a result of cell-cycle delays caused by the radiation exposure. It is possible that, under these conditions, over accumulation of cyclin B1 dilutes the mitosis-inhibitory action of the weel or other inhibitory pathways. Thus, this study presents a possible mechanism for G2/M checkpoint abrogation following ionizing radiation which may depend solely on effects associated with perturbed cell-cycle progression. PMID- 9343107 TI - Chromosomal aberrations induced in human lymphocytes by high-LET irradiation. AB - High linear energy transfer (LET) particles are more efficient than sparsely ionizing radiations in inducing chromosomal aberrations, in particular complex rearrangements. We analysed R-banded chromosome rearrangements in human lymphocytes irradiated with several ions having a wide range of LET (31.3-1435 keV/micron). The frequency of chromosome breaks unrejoined or inferred from observed rearrangements, and of complex rearrangements induced by a single particle, increased with the LET up to about 100-150 keV/micron and seemed to level off for higher LET values. Additional study was focused on damage induced by oxygen ions of three different energies. Significant cell cycle delay, and multiple chromosome rearrangements and breaks were demonstrated using Giemsa and Fluorescence-plus-Giemsa stainings, coupled with chromosome painting. Damage increased with the fluence and the LET, but at the higher LET damage decreased for fluences > 10(7) particles/cm2. Cell death and G2 block might be involved in this phenomenon. Chromosome 1 painting exhibited a high frequency of breaks and complex rearrangements, which would not have been detected using a standard staining. Complex rearrangements were induced by as few as one particle per cell nucleus and may be considered as a biological fingerprint of high-LET irradiation. PMID- 9343108 TI - G2 chromosomal radiosensitivity in fibroblasts of ataxia-telangiectasia heterozygotes and a Li-Fraumeni syndrome patient with radioresistant cells. AB - PURPOSE: To investigate whether the good discrimination we previously observed between ataxia-telangiectasia (A-T) heterozygotes and normal donors for induction of chromosome aberrations by X-rays in G2 lymphocytes is also seen in G2 fibroblasts. Also to investigate the G2 radiosensitivity of a patient with the cancer-prone Li-Fraumeni syndrome (LFS) whose fibroblasts are resistant to the lethal effects of radiation. MATERIALS AND METHODS: Fibroblasts were exposed to 0.5 Gy X-rays and harvested for metaphase analysis 90 min later. RESULTS: Four A T heterozygote cell strains were all more sensitive than seven normal controls. The LFS strain with a germline TP53 mutation was twice as sensitive as the mean control value. CONCLUSIONS: Although chromosomal, radiosensitivity is seen in A-T heterozygotes and LFS cells, the former are radiosensitive and the latter radioresistant to cell killing. Repair defects may predominate in A-T heterozygotes, inadequate genome surveillance in LFS cells. PMID- 9343110 TI - Image analysis of comet assay measurements. AB - In the last decade the 'comet assay' or 'single cell gel electrophoresis assay' has been established as a sensitive method for the detection of DNA damage and the measurement of its recovery. The results published in the literature have often been obtained with different methods for comet structure measurement. In most cases these data are not comparable with each other. Even when using similar systems for the analysis, it is difficult to obtain matching data. This presentation will describe some technical aspects of our measurement equipment and evaluation software. It focuses on necessary experimental conditions to minimize errors in obtaining such data. The software developed here allows the rapid analysis of the microscopic samples (< 2 s per image). The image analysis was designed with respect to the morphological shapes of comet cells, which were investigated with a confocal laser microscope. The system is built with standard components which are commercially available. As a measure of the amount of DNA damage the ratio of fluorescence intensity was used inside the comet tail and the fluorescence intensity of the comet head. Other parameters such as DNA content, comet area, head radius, tail length and tail moment are also determined. The reproducibility of the system has been evaluated in several experiments over a period of 5 years. PMID- 9343109 TI - Modulation of radiation-induced apoptosis by thiolamines. AB - Exposure to the thiolamine radioprotector N-(2-mercaptoethyl)-1,3-propanediamine (WR-1065) induced apoptosis in the mouse TB8.3 hybridoma after a 60-min (LD50 = 4.5 mM) or during a 20-h (LD50 = 0.15 mM) exposure. In contrast, a 20-h exposure to 17 mM L-cysteine or 10 mM cysteamine was required to induce 50% apoptosis within 20 h. Apoptosis was not induced by either a 60-min or 20-h exposure to 10 mM of the thiazolidine prodrugs ribose-cysteine (RibCys) or ribose-cysteamine (RibCyst). Thiolamine-induced apoptosis appeared to be a p53-independent process since it was induced by WR-1065 exposure in human HL60 cells. Exposure to WR-1065 (4 mM for 15 min) or cysteine (10 mM for 60 min) before and during irradiation protected cells against the induction of both DNA double-strand breaks and apoptosis, while exposure to RibCys (10 mM for 3 h) did not. Treatment with either WR-1065, cysteine, RibCys or RibCyst for 60 min beginning 60 min after irradiation did not affect the level of radiation-induced apoptosis. In contrast, treatment with either cysteine, cysteamine or RibCys for 20 h beginning 60 min after irradiation enhanced radiation-induced apoptosis. Similar experiments could not be conducted with WR-1065 because of its extreme toxicity. Our results indicate that thiolamine enhancement of radiation-induced apoptosis is not involved in their previously reported capacity to reduce radiation-induced mutations. PMID- 9343111 TI - Progression rate of radiation damage to the mouse kidney: a quantitative analysis of experimental data using a simple mathematical model of the nephron. AB - Mouse kidneys were irradiated bilaterally with a range of single or fractionated X-ray doses. After an interval of 2 weeks or 26 weeks, the animals were reirradiated with a range of single X-ray doses. The rate of development of functional kidney damage was assessed repeatedly by the 51Cr-EDTA clearance assay. The rate at which the damage is expressed was found to depend on the primary dose, on the interval between primary treatment and retreatment, and on the retreatment dose. A subset of the data was analysed using a mathematical model of nephron function. In the model, the residual activity of 51Cr-EDTA depends on the glomerular filtration rate (GFR). The GFR is related to the cellularities of three target cell populations. The filtration capacity of the glomerulus is assumed to depend on the numbers of glomerular endothelial cells and mesangial cells. The reabsorption capacity of the tubule is related to the number of tubular epithelial cells. The impact of tubulo-glomerular feedback and the reserve capacity of the kidney on residual activity is considered. The target cell populations are assumed to be of a flexible type, i.e. to consist of cells which are all both functional and self-renewing. Free parameters of the model were optimized by minimizing the residual sum of squares. With the optimized parameter values, the measured and the model-predicted rates of progression of the functional damage correspond well for a wide range of irradiation schedules. The model analysis suggests a pronounced role of tubulo-glomerular feedback in the development of functional injury in the kidney. It is concluded that the model represents a good starting point for quantitative studies of the cellular basis of radiation nephropathy. PMID- 9343112 TI - Electromagnetic millimeter waves increase the duration of anaesthesia caused by ketamine and chloral hydrate in mice. AB - BALB/c mice were injected i.p. with either ketamine 80 mg/kg or chloral hydrate 450 mg/kg. Anaesthetized mice were exposed to unmodulated electromagnetic millimeter waves at the frequency of 61.22 GHz with a peak specific absorption rate of 420 W/kg and corresponding incident power density of 15 mW/cm2 for 15 min or sham-exposed. In combination with either of the anaesthetics used, mm waves increased the duration of anaesthesia by approximately 50% (p < 0.05) in a dose (power)-dependent manner. Sham exposure to mm waves did not affect the sleeping time of mice. Pretreatment of mice with naloxone, an opioid antagonist, did not change the duration of anaesthesia caused by the corresponding chemical agent, but completely blocked or decreased the additional effect of mm waves. The data in this study indicates that exposure of mice to mm waves in vivo releases endogenous opioids or enhances the activity of opioid signalling pathway. PMID- 9343113 TI - Aging. PMID- 9343114 TI - Cerebral venous thrombosis in Behcet's disease. PMID- 9343115 TI - Cerebral infarction due to internal carotid artery dissection. PMID- 9343116 TI - Catechol-O-methyltransferase inhibition with tolcapone reduces the "wearing off" phenomenon and levodopa requirements in fluctuating parkinsonian patients. AB - BACKGROUND: More than 50% of patients with Parkinson's disease develop motor response fluctuations (the "wearing off" phenomenon) after more than five years of levodopa therapy. Inhibition of catechol-O-methyltransferase by tolcapone has been shown to increase levodopa bioavailability and plasma elimination half life, thereby prolonging the efficacy of levodopa. OBJECTIVES: The primary objective was to evaluate the efficacy of tolcapone in reducing "wearing off" in levodopa treated, fluctuating parkinsonian patients. Secondary objectives included assessment of reduction in levodopa requirements, improvement in patients' clinical status, duration of improvements, and tolerability of tolcapone. METHODS: In this multicentre, randomised, double blind, placebo controlled trial, 58 patients received placebo, 60 received 100 mg tolcapone three times daily (tid), and 59 received 200 mg tolcapone tid, in addition to levodopa/benserazide. RESULTS: After three months with 200 mg tolcapone tid, "off" time decreased by 26.2% of the baseline value, "on" time increased by 20.6% (P18; n = 32), the sensitivity of the association was 0.34, the specificity 0.93, the positive predictive value 0.85, and the negative predictive value 0.57. The diagnosic accuracy was 0.53. CONCLUSION: This association, although not sensitive, helps to select patients with high probability of Alzheimer's disease at an early stage which can be of interest for clinical and research purposes. PMID- 9343122 TI - Survey of the distribution of lesion size in multiple sclerosis: implication for the measurement of total lesion load. AB - OBJECTIVES: Quantitative measurement of lesion load on proton density or T2 weighted brain MRI in multiple sclerosis is a widely used marker of disease progression in treatment trials and natural history studies. However, it has proved difficult to obtain highly reproducible measurements. Several factors account for this, one of which is uncertainties in lesion identification, particularly very small white matter abnormalities. This paper aims to ascertain the significance of very small white matter abnormalities in the measurement of lesion load in multiple sclerosis. METHODS: All visible lesion areas identified by an experienced observer on proton density weighted spin echo brain MRI with 5 mm thick slices were measured by using a contouring technique in 15 patients with secondary progressive multiple sclerosis (SPMS) and 13 with relapsing remitting multiple sclerosis (RRMS). The size distribution of these lesions was analysed. RESULTS: 80% of the number of the lesions were smaller than 80 mm2. Lesions that were smaller than 10 mm2 (equivalent diameter <3.5 mm) made up nearly 20% of all lesions; their relative contribution to the total lesion load varied from 0.0 5.7% (mean=1.1%, median=0.65%) in individual patients, and was larger when the total lesion load was smaller (r = -0.65, P<0.001). Median lesion size was significantly smaller in the SPMS group than the RRMS group. CONCLUSIONS: The results suggest that it is prudent to identify and measure small lesions in evaluating treatment effects, and that measures are undertaken (for example, using thinner slices such as 3 mm) to improve their detection. PMID- 9343121 TI - Cerebrospinal fluid analysis differentiates between relapsing-remitting and secondary progressive multiple sclerosis. AB - OBJECTIVES: To find whether CSF analysis may differentiate between relapsing remitting and secondary progressive multiple sclerosis. METHODS: In 17 patients with relapsing-remitting and 16 patients with secondary progressive multiple sclerosis, all without current or recent relapses, albumin CSF: peripheral blood ratio, mononuclear cell number, CD4+, CD8+, and B1+ subsets, CD4+:CD8+ ratio, IgG, IgG index, IgM, IgM index, complement components C3 and C4, and C3 and C4 indices, myelin basic protein, neuron specific enolase, S100, and lactate were determined. For each measure the statistical distance measure D2 was calculated. For computation of a discriminant score variables with a P value< or =0.15 were included (two sided univariate t test). These were albumin CSF: peripheral blood ratio, mononuclear cell number, IgM, IgM index, C3, C4, neuron specific enolase, S100, and lactate. Simultaneous distributions of the variables were compared between both groups (multivariate t test) and a discriminant score was computed (linear discriminant analysis). RESULTS: The discriminant score allocated all 14 relapsing-remitting patients to the relapsing-remitting group (positive score) and 12 of 13 secondary progressive patients to the secondary progressive group (negative score). One secondary progressive patient was allocated to the relapsing-remitting group. CONCLUSIONS: Patients with relapsing-remitting or secondary progressive multiple sclerosis differ in CSF profile and CSF analysis may help to differentiate between relapsing-remitting and secondary progressive multiple sclerosis. PMID- 9343123 TI - Predictive value of clinical indices in detecting aspiration in patients with neurological disorders. AB - OBJECTIVES: (1) To evaluate the predictive value of a detailed clinical screening of aspiration in patients with neurological diseases, both with and without symptoms of dysphagia taking videofluoroscopy as the gold standard; (2) to assess the existence of risk factors for silent aspiration, measuring the cost-benefit ratio of radiological examination. METHODS: 93 consecutive patients meeting the diagnostic criteria for a neurological disease with a risk of swallowing dysfunctions (cerebrovascular accidents, brain injury, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, myotonic dystrophy, and abiotrophic diseases) underwent a detailed clinical assessment using a 25 item form to check for symptoms of dysphagia and impairment of the oropharyngeal swallowing mechanism. The 3 oz water swallow test was also performed to assess the aspiration risk. Sensitivity, specificity, positive predictive, and negative predictive values (NPV) of dysphagia, history of cough on swallowing, and 3 oz test positivity, versus videofluoroscopy documented aspiration, taken as the gold standard, were measured in all the patients and in subgroups with different neurological disorders. RESULTS: Non-specific complaints of dysphagia showed a very poor predictive value, whereas the symptom "cough on swallowing" proved to be the most reliable in predicting the risk of aspiration, with 74% sensitivity and specificity, 71% positive predictive, and 77% negative predictive value. The standardised 3-oz test had a higher predictive potential than the clinical signs, but had low sensitivity. The association of cough on swallowing with the 3 oz test gave a positive predictive of 84%, and an negative predictive value of 78%. In cases where the clinical tests failed to detect any impairment, videofluoroscopy documented only a low risk (20%) for mild aspiration. CONCLUSIONS: The association of two clinical items (such as history of cough on swallowing and 3 oz test positivity) provides a useful screening tool, the cost:benefit ratio of which seems very competitive in comparison with videofluoroscopy in aspiration risk evaluation. PMID- 9343124 TI - Bilateral reductions in hippocampal volume in adults with epilepsy and a history of febrile seizures. AB - OBJECTIVES: To examine the degree and frequency of reductions in hippocampal volume in patients with temporal lobe epilepsy with and without a history of febrile seizures. METHODS: In vivo measures of hippocampal volume were computed from three dimensional gradient echo (FLASH) images in 44 patients undergoing comprehensive evaluations for epilepsy surgery. Twenty one patients (48%) reported a history of febrile seizures. The volumes from these patients were compared with those from 23 patients without a history of febrile seizures and 34 healthy controls. RESULTS: The febrile seizure group had significant reductions in volume, both ipsilateral (30% decrease) and contralateral (15% decrease), to the EEG seizure focus. Twelve of 18 patients with febrile seizures exhibited clinically significant ipsilateral volume reductions, defined as volumes falling 2 SD below the mean obtained from the control sample. Only four of 19 patients without febrile seizures exhibited this degree of reduction. No significant correlations were found between seizure variables (for example, duration of epilepsy, seizure frequency) and ipsilateral reductions in volume. However, a significant inverse correlation (r=-0.45, P<0.05) between seizure frequency and the volume of the hippocampus contralateral to the seizure focus was found in the febrile seizure group. CONCLUSION: These results suggest that a history of febrile seizures is associated with the finding of a smaller hippocampus on the side ipsilateral to the subsequent temporal lobe focus whereas chronic factors seem to be be related to pathology contralateral to the seizure focus. PMID- 9343126 TI - Botulinum A toxin as a treatment of detrusor-sphincter dyssynergia in patients with spinal cord injury: MRI controlled transperineal injections. AB - OBJECTIVES: To correlate clinical and urodynamic findings with MRI in patients with spinal cord injury and detrusor-sphincter dyssynergia who were consecutively treated with transperineal injections of botulinum-A toxin (BTX-A) under EMG control. METHODS: Six patients with spinal cord injury and upper motor neuron bladder dysfunction associated with detrusor-sphincter dyssynergia were prospectively analysed. One hundred international units (IU) BTX-A (Botox in 1 ml normal saline without preservative) diluted 1 to 1 with 1 ml gadopentetate were injected transperineally under EMG control. MRI was started immediately after needle withdrawal. RESULTS: In all six patients gadopentetate was located in the external urethral sphincter on MRI. In no patient did traces of gadopentetate appear in the perineal musculature located in the vicinity of the external urethral sphincter. No patient developed resistance to BTX-A. All patients showed an (ongoing) improvement of their voiding function after BTX-A injections. CONCLUSIONS: Transperineal injections of BTX-A under EMG control are efficient in the release or amelioration of lower urinary tract obstruction due to detrusor sphincter dyssynergia in patients with spinal cord injury. Despite well described methods, EMG of the external urethral sphincter is difficult and it is not possible to definitively exclude false recordings of the surrounding perineal musculature. By the use of MRI it was shown that both the EMG recordings and transperineal injection method are precise. PMID- 9343125 TI - Effects of antipsychotic medication on electromyographic responses to transcranial magnetic stimulation of the motor cortex in schizophrenia. AB - OBJECTIVE: To assess the effect of antidopaminergic antipsychotic medication on the electromyographic (EMG) responses of thenar muscles to transcranial magnetic stimulation (TMS) of the motor cortex in schizophrenic patients. METHODS: A group of nine drug naive schizophrenic patients was compared with a group of nine schizophrenic patients established on neuroleptic medication. Surface EMG recordings were made from the thenar muscles while patients maintained a weak isometric voluntary contraction. TMS was applied using a 9 cm circular stimulating coil centred over the vertex. The EMG responses to up to 50 magnetic stimuli were rectified and averaged. RESULTS: There was no difference in threshold TMS strength for eliciting compound motor evoked potentials (cMEPs), or in their latency, in drug naive and medicated patients. In some patients the silent period (SP) was clearly made up of two parts and the percentage of control levels of voluntary EMG was measured in each component. During the early component of the SP there was a weaker (P<0.05) suppression of EMG in the medicated patients (mean 73.9 (SEM) 5.5% of control levels) compared with the drug naive patients (54.7 (SEM) 7.3% of control levels). This resulted in the latency of maximum suppression of voluntary EMG being longer (P<0.05) in the medicated patients (38.3 (2.4) ms) than in the drug naive patients (28.2 (0.7) ms). During the late component of the SP voluntary EMG was reduced to similar levels (P>0.05) in both medicated (48.2 (7.7)% of control levels) and drug naive (58 (7.8)% of control levels) patients. CONCLUSION: The results are discussed with reference to the disrupted inhibition seen in the early part of the SP in Parkinson's disease and drug induced parkinsonism. The future uses of motor responses to TMS as a marker for the status of antipsychotic medication are considered. PMID- 9343127 TI - Proton MRS and quantitative MRI assessment of the short term neurological response to antiretroviral therapy in AIDS. AB - OBJECTIVE: To investigate MRI and proton spectroscopy changes in five patients with HIV associated dementia complex (HADC) treated with antiretroviral therapy. METHODS: Three markers were evaluated: (1) CSF/intracranial volume ratio; (2) T2 weighted signal ratio between parieto-occipital white and subcortical grey matter; and (3) metabolite ratios from long echo time (TE=135 ms) single voxel proton spectra of parieto-occipital white matter. RESULTS: Spectroscopic changes indicated initial increases in N-acetyl/(N-acetyl + choline + creatine) ratio (NA/(NA+ Cho+Cr)) and progression of atrophy after initiation of antiretroviral therapy in four of five patients. When the neurological status of the patients subsequently deteriorated (two of five patients), the NA/(NA+Cho+Cr) ratio also declined. CONCLUSIONS: Spectroscopic changes mirror reversible neuronal dysfunction. These objective, non-invasive techniques may be used for monitoring the neurological effects of antiretroviral drug therapy in patients with HADC. PMID- 9343128 TI - Left of what? The role of egocentric coordinates in neglect. AB - OBJECTIVES: Egocentric coordinate systems centred on the trunk, head, and gaze have been investigated in a patient who displays severe extrapersonal neglect and in five control subjects. METHODS: The subjects were tested with a blind tactile exploration task in five different experimental conditions in which the role of the three distinct frames of reference was individually controlled. RESULTS: Only the trunk centred coordinates significantly influenced the performance of the patient, therefore proving of paramount importance in determining the boundaries of the neglected field. Similar results emerged from a single word reading task, in which the patient's performance improved when the stimuli were presented to the right of his body's midline. CONCLUSION: These findings point to the importance of the body centred coordinate system in determining the area of extrapersonal spatial neglect. PMID- 9343129 TI - Timing of surgery in patients with aneurysmal subarachnoid haemorrhage: rebleeding is still the major cause of poor outcome in neurosurgical units that aim at early surgery. AB - OBJECTIVE: To investigate prospectively the proportion of patients actually operated on early in units that aim at surgery in the acute phase of aneurysmal subarachnoid haemorrhage (SAH) and what is the main current determinant of poor outcome. METHODS: A prospective analysis of all SAH patients admitted during a one year period at three neurosurgical units that aim at early surgery. The following clinical details were recorded: age, sex, date of SAH, date of admission to the neurosurgical centre, whether a patient was referred by a regional hospital or a general practitioner, Glasgow coma scale and grade of SAH (World Federation of Neurological Surgeons (WFNS) score) on admission at the neurosurgical unit, results of CT and CSF examination, the presence of an aneurysm on angiography, details of treatment with nimodipine or antifibrinolytic agents, and the date of surgery to clip the aneurysm. At follow up at three months, the patients' clinical outcome was determined with the Glasgow outcome scale and in cases of poor outcome the cause for this was recorded. RESULTS: The proportion of patients that was operated on early--that is, within three days after SAH--was 55%. Thirty seven of all 102 admitted patients had a poor outcome. Rebleeding and the initial bleeding were the main causes of this in 35% and 32% respectively of all patients with poor outcome. CONCLUSIONS: In neurosurgical units with what has been termed "modern management" including early surgery, about half of the patients are operated on early. Rebleeding is still the major cause of poor outcome. PMID- 9343130 TI - Guillain-Barre syndrome in Taiwan: a clinical study of 167 patients. AB - OBJECTIVE: To identify clinical characteristics of various forms of Guillain Barre syndrome in Taiwan. METHODS: The clinical and electrophysiological data of 167 consecutive patients with Guillain-Barre syndrome admitted to Chang Gung Memorial Hospital, a general paediatric and adult hospital in Taiwan, were reviewed. RESULTS: Analysis of age distribution disclosed a high incidence (21%) among patients under the age of 10 years. Seasonal preponderance in Spring (March to May) was found. Utilizing clinical and electrophysiological data, these 167 patients with Guillain-Barre syndrome were subclassified; 82 (49%) had acute inflammatory demyelinating polyradiculoneuropathy (AIDP), 32 (19%) had Fisher syndrome (FS), and six (4%) had axonal forms of Guillain-Barre syndrome. The remaining 47 (28%) patients were unclassified. Patients with AIDP and FS had many common clinical features, including seasonal distribution, history of preceding illness, sensory abnormalities, cranial nerve involvement except for extraocular motor nerves, and albuminocytological dissociation on examination of CSF. Follow up study on 145 patients disclosed that 127 (87%) recovered satisfactorily, 14 (10%) were persistently disabled, and four (3%) died during admission to hospital. Clinical features associated with poor outcome (persistent disability or death) were requirement for mechanical ventilation, a low mean compound muscle action potential amplitude (< or = 10% of the lower limit of normal), and age greater than 40 years. CONCLUSION: Guillain-Barre syndrome in Taiwan showed a peculiar age and seasonal distribution and a high frequency of FS not seen in other series. Given that patients with AIDP and FS had many common clinical features, AIDP and FS may have similar underlying pathological mechanisms. PMID- 9343131 TI - Human second somatosensory area: subdural and magnetoencephalographic recording of somatosensory evoked responses. AB - OBJECTIVE: To investigate somesthetic functions of the perisylvian cortex. METHODS: Somatosensory evoked magnetic fields (SEFs) and somatosensory evoked potentials (SEPs) of the perisylvian cortex were recorded directly from subdural electrodes in a patient with a left frontal brain tumour. RESULTS: The most prominent SEP components after electrical stimulation of the right and left hands and the right foot were double peaked negativity recorded just above the sylvian fissure (latency 80 to 150 ms), respectively (N1a and N1b). Generator sources for the magnetoencephalographic counterparts of those peaks (N1a(m) and N1b(m)) were both localised at the upper bank of the sylvian fissure, and those of N1a(m) were more anteromedially located than those of N1b(m). CONCLUSIONS: These findings suggest the existence of at least two separate somatosensory areas within the human perisylvian cortex. PMID- 9343132 TI - Cerebrospinal fluid transthyretin: aging and late onset Alzheimer's disease. AB - The deposition of insoluble beta-amyloid protein fibrils is probably the central event in the pathogenesis of Alzheimer's disease. Cerebrospinal fluid inhibits this fibril formation, likely by the intervention of one or several proteins binding to soluble beta-amyloid protein. In vitro, transthyretin (TTR), a CSF protein, impedes amyloid fibrillogenesis. Lowered concentrations of CSFTTR could therefore be associated with Alzheimer's disease. Concentrations of TTR in CSF samples from 149 consecutive patients were assayed, using a kinetic nephelemetric method. These concentrations were correlated positively with age, but were significantly lower in patients with Alzheimer's disease. These data raise the possibility that amyloid fibril formation could be promoted in patients with late onset Alzheimer's disease by the lack of sufficient concentrations of TTR. PMID- 9343133 TI - Left ventricular dysfunction: a clue to cognitive impairment in older patients with heart failure. AB - OBJECTIVES: Cognitive impairment has been reported in middle aged patients with end stage heart failure. This cross sectional study assessed the prevalence and determinants of cognitive dysfunction in older patients with mild to moderate heart failure. METHODS: 57 consecutive patients (mean age 76.7 years) with chronic heart failure underwent physical examination, blood chemistry, urinalysis, chest radiography ECG, Doppler echocardiography, and the mini mental state examination (MMSE), mental deterioration battery, depression scale of the Center for Epidemiological Studies (CES-D), Katz activities of daily living, and instrumental activities of daily living 24 hours before hospital discharge. RESULTS: MMSE scores <24 were found in 53% of participants. The MMSE score was associated with left ventricular ejection fraction according to a non-linear correlation, so that cognitive performance was significantly lower in subjects with left ventricular ejection fraction < or =30%. The same pattern of correlation was evidenced between left ventricular ejection fraction and both the attention sub-item of MMSE and the Raven test score. In a multivariate linear regression model, after adjusting for age, sex, and a series of clinical data and objective tests, both age (beta=-0.30; P=0.038) and the natural log of left ventricular ejection fraction (beta=0.58; P=0.001) were associated with the MMSE score. CONCLUSION: Cognitive impairment in older patients with chronic heart failure is common, and independently associated with lower left ventricular ejection fraction. Given the overwhelming incidence and prevalence of heart failure in older populations, early detection of cognitive impairment in these subjects with prompt, intensive treatment of left ventricular systolic dysfunction may prevent or delay a remarkable proportion of dementia in advanced age. PMID- 9343134 TI - Corpora amylacea in hippocampal sclerosis. AB - Corpora amylacea have been reported in around 60% of hippocampal sclerosis specimens. The aim was to determine whether there are clinical and quantitative hippocampal MRI differences between hippocampal sclerosis with and without corpora amylacea. Corpora amylacea density was determined in 46 resected hippocampi of patients with temporal lobe epilepsy, using a three dimensional microscopical counting technique. Forty one hippocampi had hippocampal sclerosis. Twenty six of the 41 (63%) hippocampal sclerosis specimens contained corpora amylacea, which were found in highest numbers in the CA1 subregion of the hippocampus. Corpora amylacea density in the CA1 correlated inversely with the neuronal density in CA1. Hippocampal sclerosis with corpora amylacea had the same clinical and quantitative hippocampal MRI characteristics as hippocampal sclerosis without corpora amylacea, and did not affect seizure outcome after surgery adversely. In conclusion, formation of corpora amylacea seems to be a pathological response to neuronal cell loss in most hippocampal sclerosis specimens, with no clear clinical and quantitative hippocampal MRI correlates. PMID- 9343135 TI - A study on a new antineural antibody in a case of paraneoplastic sensory neuropathy associated with breast carcinoma. AB - Paraneoplastic sensory neuropathy is a remote effect of cancer, usually associated with small cell lung carcinoma and anti-Hu antibody. This report details the case of a 59 year old woman with a breast carcinoma and a paraneoplastic sensory neuropathy characterised by chronic asymmetric sensory neuropathy. Anti-Hu antibody was not detected in her serum; nor were other known antineuronal antibodies such as anti-Ri and Yo. However, we have found an antineural antibody that reacted to a 106 kDa mouse neural antigen which has not yet been reported. Immunohistochemically, this antineural antibody bound to the posterior grey horn. This finding suggests that this antineural antibody may play an important part in the pathogenesis of the sensory neuropathy of this patient. PMID- 9343137 TI - Of insects and eggs: a case report. AB - A middle aged woman presented with delusions of infestation and multimodal hallucinations due to an underlying glioma of the corpus callosum. After surgery, the phenomena in question changed and finally disappeared. A recurrence of the tumour caused dementia. PMID- 9343136 TI - The case of the disappearing glioma. AB - A case of glioblastoma multiforme disappearing on CT after a course of dexamethasone is reported. The possibility of mistaking this for lymphoma is discussed. There is no other case similar to this described in the literature. PMID- 9343138 TI - Panencephalopathic type of Creutzfeldt-Jakob disease associated with cadaveric dura mater graft. AB - A 52 year old man with Creutzfeldt-Jakob disease who received a cadaveric dura mater graft 99 months before the onset is reported. The prion protein gene was homozygous for methionine at the polymorphic codon 129. Neuropathological examination disclosed a panencephalopathic type of Creutzfeldt-Jakob disease which was characterised by severe involvement of the cerebral white matter and cerebellum, as well as of the cerebral cortical and deep grey matter. Thus the panencephalopathic type of Creutzfeldt-Jakob disease may occur in association with cadaveric dura mater grafts. PMID- 9343139 TI - Blink induced centrotemporal spikes in benign childhood epilepsy with centrotemporal spikes. AB - A 10 year old girl with benign childhood epilepsy with centrotemporal spikes showed centrotemporal spikes induced by blinking even in a dark room. Spikes could not be induced by photic stimulation, eye closure, eye movement, eye deviation, or passive blinks. There have been no previous reports of spikes induced by blinks in benign childhood epilepsy with centrotemporal spikes. PMID- 9343140 TI - Ischaemic myelopathy associated with cocaine: clinical, neurophysiological, and neuroradiological features. AB - Two patients with spinal infarction and one patient with the previously unreported complication of spinal transient ischaemic attack associated with cocaine misuse are reported. Spinal MRI documented an infarction in the territory of the anterior spinal artery in the first two patients and was completely normal in the patient with a transient ischaemic attack. Motor evoked potentials were abnormal in all three patients. PMID- 9343141 TI - Machado-Joseph disease presenting as severe asymmetric proximal neuropathy. AB - Despite much effort, a 74 year old man with progressive proximal weakness and sensory disturbances due to axonal neuropathy remained a diagnostic problem. Investigation of his family disclosed an additional patient with a cerebellar syndrome and a family member with mainly pyramidal features. Analysis of DNA showed a CAG repeat expansion in the Machado-Joseph disease gene in all three patients. Although not conclusively proved, we think that the neuropathy of the index case is linked to the CAG repeat expansion. Machado-Joseph disease should be considered in progressive axonal neuropathy. PMID- 9343142 TI - Skin sympathetic nerve activity in Guillain-Barre syndrome: a microneurographic study. AB - To assess autonomic dysfunction, skin sympathetic nerve activity (SSNA) of four patients with Guillain-Barre syndrome was microneurographically studied in the acute and remission phase. Autonomic symptoms such as sinus tachycardia, palmar hyperhidrosis, hypertension, and orthostatic hypotension were present in the acute phase, but all subsided during remission. Basal resting SSNA and the responses to various physical and mental stimuli were all increased in the acute phase and returned almost to normal during remission. Rate of response in sweat rate and blood flow against SSNA were kept proportionally constant during both the acute and remission phases. These findings suggest that some autonomic nerve symptoms of Guillain-Barre syndrome, particularly during the acute phase, are due to increased SSNA. PMID- 9343143 TI - Median nerve injury: an underrecognised complication of brachial artery cardiac catheterisation? AB - OBJECTIVE: To describe the local neurological complications associated with cardiac catheterisation via the right brachial artery. METHODS: A follow up study to determine the mechanism of injury and outcome of patients who sustained a high median nerve palsy after this procedure. Five right handed patients were identified in a 24 month period. Each was assessed clinically and electrophysiologically at presentation. All were followed up initially (range six to 22 months) clinically, electrophysiologically, and using components from the Chessington occupational therapy neurological assessment battery (COTNAB) functional hand assessment. RESULTS: The incidence of this complication was between 0.2 and 1.4%. Three mechanisms of injury were identified. These included direct nerve compression due to formation of antecubital fossa haematoma, direct nerve trauma, and ischaemia secondary to brachial artery occlusion. The initial neurological and nerve conduction deficits improved with time. However, all cases had persistent disability in hand function as documented clinically and on the dexterity and stereognosis subcomponent of the COTNAB test. CONCLUSION: This is an uncommon, but probably underrecognised complication. Those performing cardiac catheterisation via the right brachial artery should be aware of the potential risks of damage to the median nerve. They should evaluate hand function after the procedure and take prompt action if median nerve dysfunction is noted. Damage to the median nerve results in appreciable long term disability, which may have medicolegal relevance. PMID- 9343144 TI - Moclobemide and selegeline in the treatment of depression in Parkinson's disease. PMID- 9343145 TI - Brain biopsy and patients with atypical presentations of sporadic Creutzfeldt Jakob disease. PMID- 9343146 TI - Transient semantic amnesia: a new syndrome? PMID- 9343147 TI - Alopecia with carbamazepine in two patients with focal seizures. PMID- 9343149 TI - Successful outcome with aggressive treatment of acute haemorrhagic leukoencephalitis. PMID- 9343148 TI - Neurological manifestations of coeliac disease. PMID- 9343150 TI - Adult Niemann-Pick disease type C mimicking features of multiple sclerosis. PMID- 9343151 TI - Motor cortex stimulation does not reset primary orthostatic tremor. PMID- 9343152 TI - Frozen section in pituitary surgery. PMID- 9343153 TI - Bilateral posteroventral pallidotomy for severe antipsychotic induced tardive dyskinesia and dystonia. PMID- 9343154 TI - Pretarsal injections of botulinum toxin improve blephospasm in previously unresponsive patients. PMID- 9343155 TI - Rehabilitation of gait in Parkinson's disease. PMID- 9343156 TI - Association between HIV distal symmetric polyneuropathy and Mycobacterium avium complex infection. PMID- 9343157 TI - Primer tRNAs for reverse transcription. PMID- 9343158 TI - In vitro binding of purified murine ecotropic retrovirus envelope surface protein to its receptor, MCAT-1. AB - An amino-terminal portion of the Friend murine leukemia virus (MLV) envelope surface protein [SU, residues 1 to 236 [SU:(1-236)]] and its receptor, MCAT-1, were each purified from insect cells after expression by using recombinant baculoviruses. Friend SU:(1-236) bound specifically to Xenopus oocytes that expressed MCAT-1 with an affinity (Kd, 55 nM) similar to that of viral SU binding to permissive cells. Direct binding of Friend SU:(1-236) to purified MCAT-1 was observed in detergent and after reconstitution into liposomes. Analysis of binding demonstrated that MCAT-1 and Friend SU:(1-236) interact with a stoichiometry of near 1:1. These findings demonstrate that the amino-terminal domain from the SU of ecotropic murine retroviruses contains an MCAT-1 binding domain. PMID- 9343159 TI - Identification of envelope protein residues required for the expanded host range of 10A1 murine leukemia virus. AB - The 10A1 murine leukemia virus (MuLV) is a recombinant type C retrovirus isolated from a mouse infected with amphotropic MuLV (A-MuLV). 10A1 and A-MuLV have 91% amino acid identity in their envelope proteins yet display different host ranges. For example, CHO-K1 cells are resistant to A-MuLV but susceptible to infection by 10A1. We have now determined that retroviral vectors bearing altered A-MuLV envelope proteins containing 10A1-derived residues at positions 71 (A71G), 74 (Q74K), and 139 (V139M) transduce CHO-K1 cells at efficiencies similar to those achieved with 10A1 enveloped vectors. A-MuLV enveloped retroviral vectors with these three 10A1 residues were also able to transduce A-MuLV-infected NIH 3T3 cells. This observation is consistent with the ability of vectors bearing this altered A-MuLV envelope protein to recognize the 10A1-specific receptor present on NIH 3T3 cells and supports the possibility that residues at positions 71, 74, and 139 of the 10A1 envelope SU protein account for the expanded host range of 10A1. PMID- 9343160 TI - Immunoglobulin G is the main protective antibody in mouse vaginal secretions after vaginal immunization with attenuated herpes simplex virus type 2. AB - We investigated the protective role of antibodies in vaginal secretions of mice that were immune to vaginal challenge with herpes simplex virus type 2 (HSV-2). Unfractionated vaginal immunoglobulins from immune and nonimmune mice and affinity-purified immunoglobulin G (IgG) and secretory IgA (S-IgA) from immune secretions were adjusted to their concentrations in vivo. Wild-type HSV-2 was incubated in the immunoglobulin preparations for 15 min in vitro, followed by inoculation into vaginae of nonimmune mice. HSV-2 was neutralized by unfractionated antibody and purified IgG from immune secretions but not by unfractionated nonimmune antibody or by purified immune S-IgA. The protective effect of IgG in vivo was investigated by passively transferring purified serum IgG from immune and nonimmune donors to nonimmune recipients before vaginal challenge infection. Immune IgG significantly reduced the percentage of vaginal epithelium infected, concentrations of shed virus protein in the vaginal lumen, and illness scores, even though the viral antibody titers in serum and vaginal secretions of recipient mice at the time of challenge were only 29 and 8%, respectively, of those in actively immunized mice. Additionally, removal of vaginal secretions from immune mice 10 min before vaginal challenge with HSV-2 significantly increased the concentration of shed virus protein in the vaginal lumen after challenge. Collectively, the data indicate that IgG antibody in vaginal secretions of immune mice provides early protection against vaginal challenge infection, probably by neutralizing virus in the vaginal lumen. In contrast, S-IgA antibody contributed relatively little to immune protection of the vagina. PMID- 9343161 TI - Three distinct envelope domains, variably present in subgroup B feline leukemia virus recombinants, mediate Pit1 and Pit2 receptor recognition. AB - Subgroup B feline leukemia viruses (FeLV-Bs) evolve from subgroup A FeLV (FeLV-A) by recombining with portions of endogenous FeLV envelope sequences in the cat genome. The replication properties of FeLV-B are distinct from those of FeLV-A; FeLV-B infects many nonfeline cell lines and recognizes the human Pit1 (HuPit1) receptor, whereas FeLV-A infects primarily feline cells, using a distinct but as yet undefined receptor. Here, we demonstrate that some FeLV-Bs can also use human Pit2 (HuPit2) and hamster Pit2 (HaPit2) for entry. By making viruses that contain chimeric surface (SU) envelope proteins from FeLV-A and FeLV-B, and testing their infectivity, we have defined genetic determinants that confer host range and specific receptor recognition. HuPit1 receptor recognition determinants localize to the N-terminal region of the FeLV-B SU, amino acids 83 to 116, encompassing the N-terminal portion of variable region A (VRA). While this 34-amino-acid domain of the FeLV-B VRA is sufficient for infection of some cells (feline, canine, and human), amino acids 146 to 249 of FeLV-B, which include variable region B (VRB), were required for efficient infection in other cell types (hamster, bovine, and rat). Chimeras encoding FeLV-B VRA and VRB also infected cells expressing HaPit2 and HuPit2 receptors more efficiently than chimeras encoding only the VRA of FeLV-B, suggesting that VRB provides a secondary determinant that is both cell and receptor specific. However, viruses containing additional FeLV-B sequences in the C terminus of SU could not recognize HuPit2, implying that there is a determinant beyond VRB that negatively affects HuPit2 interactions. Thus, Pit2 recognition may drive selection for the generation of specific FeLV-B recombinants, offering an explanation for the two major classes of FeLV-B that have been observed in vivo. Furthermore, the finding that some FeLV-Bs can use both Pit1 and Pit2 may explain previous observations that FeLV-B and GALV, which primarily uses Pit1, display nonreciprocal interference on many cell types. PMID- 9343162 TI - A mutation in integrase can compensate for mutations in the simian immunodeficiency virus att site. AB - Sequences at the left terminus of U3 in the left long terminal repeat (LTR) and at the right terminus of U5 in the right LTR are important for integration of retroviral DNA. In the infectious pathogenic molecular clone of simian immunodeficiency virus strain mac239 (SIVmac239), 10 of the 12 terminal base pairs form an imperfect inverted repeat structure (5' TGGAAGGGATTT 3' [nucleotides 1 to 12] and 3' ACGATCCCTAAA 5' [nucleotides 10279 to 10268]). Nineteen different mutant forms of SIVmac239 proviral DNA with changes at one or more of the positions in each of the 12-terminal-base-pair regions were constructed. Viral replication was severely or completely compromised with nine of these mutants. Revertants appeared 40 to 50 days after transfection in two independent experiments with mutant 7, which contained changes of AGG to TAC at positions 5 to 7 in U3 and TCC to GAA at positions 10275 to 10273 in U5. Virus produced at these times from mutant 7 transfection replicated upon reinfection with only a slight delay when compared to the wild type. Sequence analysis of the LTR and integrase regions from infected cultures revealed two predominant changes: G to A at position 10275 in U5 and Glu to Lys at position 136 in integrase. Derivatives of clone 7 in which these changes were introduced individually and together were constructed by site-specific mutagenesis. Each change individually restored replication capacity only partially. However, the combination of both mutations restored replicative capacity to that of the original revertants. These results indicate that changes in integrase can compensate for mutations in the terminal nucleotides of the SIV LTR. The results further indicate that resistance to integrase inhibitors may include both integrase and LTR mutations. PMID- 9343163 TI - Molecular analysis of an enhancin gene in the Lymantria dispar nuclear polyhedrosis virus. AB - A Lymantria dispar nuclear polyhedrosis virus (LdMNPV) gene has been identified that encodes a homolog to the granulovirus (GV) enhancin proteins that are capable of enhancing the infection of other baculoviruses. Enhancin genes have been identified and sequenced for three species of GVs but have not been found in any other nuclear polyhedrosis virus to date. The LdMNPV enhancin gene is located between 67.6 and 70.1 kbp on the viral genome. Northern and primer extension analyses of viral RNAs indicate that the enhancin gene transcripts are expressed at late times postinfection from a consensus baculovirus late promoter. The LdMNPV enhancin exhibits 29% amino acid identity to the enhancin proteins of the Trichoplusia ni, Pseudaletia unipuncta, and Helicoverpa armigera GVs. All four proteins contain a conserved zinc-binding domain characteristic of metalloproteases. A recombinant virus (enhancin::cat) was constructed in which the LdMNPV enhancin gene was inactivated by insertion mutagenesis in order to ascertain the effect of the enhancin protein on viral potency. The bioassay results indicate that disruption of the enhancin gene in the LdMNPV results in a reduction in viral potency. PMID- 9343164 TI - Live, attenuated simian immunodeficiency virus vaccines elicit potent resistance against a challenge with a human immunodeficiency virus type 1 chimeric virus. AB - Three rhesus macaques, previously immunized with SIVdelta3 or SIVdelta2, each an attenuated derivative of SIVmac239, and two naive monkeys were challenged with 30,000 50% tissue culture infective doses of SHIV, an SIV/human immunodeficiency virus type 1 (HIV-1) chimeric virus bearing the dual-tropic envelope of HIV 1DH12. By several criteria, including virus isolation, serological assays, and PCR (both DNA and reverse transcriptase), SHIV levels were reduced to barely detectable levels in the circulating blood of vaccinated animals. The resistant SIV-vaccinated macaques had no preexisting neutralizing antibodies directed against SHIV, nor did they produce neutralizing antibodies at any time over a 14 month observation period following SHIV challenge. Interestingly, SIV sequences, derived from the vaccine, could be amplified from numerous tissue samples collected at the conclusion of the experiment, 60 weeks postchallenge, but SHIV specific sequences (viz., HIV-1 env) could not. These results demonstrate that live attenuated SIV vaccines provide strong long-term protection even against challenge strains with highly divergent envelope sequences. PMID- 9343165 TI - Mutation of Tp53 contributes to the malignant phenotype of Abelson virus transformed lymphoid cells. AB - Abelson murine leukemia virus transforms pre-B cells in vitro and induces rapid onset pre-B-cell lymphoma in vivo. Expression of an active v-Abl protein tyrosine kinase is required for the oncogenic functions of the virus. Despite the strong growth-stimulatory signal provided by v-Abl, the virus-induced tumors are clonal or oligoclonal, and changes in the growth and oncogenic potential of in vitro transformants occur during the derivation of the cell lines. Both of these features suggest that v-Abl expression must be complemented by changes in expression of one or more cellular genes for cells to acquire a fully malignant phenotype. Such genes could include other oncogenes or tumor suppressor genes. Among the latter is Tp53, a gene mutated in many spontaneous cancers. To determine if mutation of the Tp53 tumor suppressor gene plays a role in Abelson virus transformation, conformation-specific monoclonal antibodies were used to examine p53 expression in a panel of Abelson virus-transformed pre-B cells. Expression of mutant forms of p53 was detected in over 40% of the isolates. Sequence analysis revealed the presence of point mutations affecting the highly conserved central portion of the protein. These mutations interfered with the ability of p53 to activate transcription from a promoter containing p53 responsive elements and to induce apoptosis in response to DNA damage. In addition, cells expressing mutant forms of p53 induced a higher frequency of tumors with a more rapid course compared to transformants expressing wild-type p53. These data suggest that Tp53 is one important cellular gene involved in malignant transformation by Abelson virus. PMID- 9343166 TI - A simian virus 40 large T-antigen segment containing amino acids 1 to 127 and expressed under the control of the rat elastase-1 promoter produces pancreatic acinar carcinomas in transgenic mice. AB - The simian virus 40 large T antigen induces tumors in a wide variety of tissues in transgenic mice, the precise tissues depending on the tissue specificity of the upstream region controlling T-antigen expression. Expression of mutant T antigens that contain a subset of the protein's activities restricts the spectrum of tumors induced. Others showed previously that expression of a mutant large T antigen containing the N-terminal 121 amino acids (T1-121) under control of the lymphotropic papovavirus promoter resulted in slow-growing choroid plexus tumors, whereas full-length T antigen under the same promoter induced rapidly growing CPR tumors, T-cell lymphomas, and B-cell lymphomas. In those instances, the alteration in tumor induction or progression correlated with inability of the mutant large T antigen to bind the tumor suppressor p53. In the study reported here, we investigated the capacity of an N-terminal T antigen segment (T1-127) expressed in conjunction with small t antigen under control of the rat elastase-1 (E1) promoter to induce pancreatic tumors. The results show that pancreases of transgenic mice expressing T1-127 and small t antigen display acinar cell dysplasia at birth that progresses to neoplasia. The average age to death in these mice is within the range reported for transgenic mice expressing full length T antigen under control of the E1 promoter. These results indicate that sequestering p53 by binding is not required for the development of rapidly growing acinar cell carcinomas. In addition, we provide evidence that small t antigen is unlikely to be required. Finally, we show that the p53 protein in acinar cell carcinomas is wild type in conformation. PMID- 9343167 TI - Phosphorylation within the amino-terminal acidic domain I of the phosphoprotein of vesicular stomatitis virus is required for transcription but not for replication. AB - Phosphorylation by casein kinase II at three specific residues (S-60, T-62, and S 64) within the acidic domain I of the P protein of Indiana serotype vesicular stomatitis virus has been shown to be critical for in vitro transcription activity of the viral RNA polymerase (P-L) complex. To examine the role of phosphorylation of P protein in transcription as well as replication in vivo, we used a panel of mutant P proteins in which the phosphate acceptor sites in domain I were substituted with alanines or other amino acids. Analyses of the alanine substituted mutant P proteins for the ability to support defective interfering RNA replication in vivo suggest that phosphorylation of these residues does not play a significant role in the replicative function of the P protein since these mutant P proteins supported replication at levels > or = 70% of the wild-type P protein level. However, the transcription function of most of the mutant proteins in vivo was severely impaired (2 to 10% of the wild-type P-protein level). The level of transcription supported by the mutant P protein (P(60/62/64)) in which all phosphate acceptor sites have been mutated to alanines was at best 2 to 3% of that of the wild-type P protein. Increasing the amount of P(60/62/64) expression in transfected cells did not rescue significant levels of transcription. Substitution with other amino acids at these sites had various effects on replication and transcription. While substitution with threonine residues (P(TTT)) had no apparent effect on transcription (113% of the wild-type level) or replication (81% of the wild-type level), substitution with phenylalanine (P(FFF)) rendered the protein much less active in transcription (< 5%). Substitution with arginine residues led to significantly reduced activity in replication (6%), whereas glutamic acid substituted P protein (P(EEE)) supported replication (42%) and transcription (86%) well. In addition, the mutant P proteins that were defective in replication (P(RRR)) or transcription (P(60/62/64)) did not behave as transdominant repressors of replication or transcription when coexpressed with wild-type P protein. From these results, we conclude that phosphorylation of domain I residues plays a major role in in vivo transcription activity of the P protein, whereas in vivo replicative function of the protein does not require phosphorylation. These findings support the contention that different phosphorylated states of the P protein regulate the transcriptase and replicase functions of the polymerase protein, L. PMID- 9343169 TI - Differential effects of the splice acceptor at nucleotide 3295 of human papillomavirus type 31 on stable and transient viral replication. AB - In human papillomavirus type 31 (HPV-31), the E1--E4 and E5 open reading frames are expressed from polycistronic mRNAs. The major polycistronic mRNAs which encode E1--E4 and E5 are spliced messages which utilize a splice acceptor at nucleotide (nt) 3295 (SPA3295). Our laboratory recently developed a recombinant system for the synthesis of HPVs following immortalization of primary keratinocytes with cloned HPV-31 genomes (M. G. Frattini et al., Proc. Natl. Acad. Sci. USA 93:3062-3067, 1996). These immortalized cell lines are capable of maintaining HPV-31 DNA as episomes and induce the synthesis of virions in organotypic raft culture. In this study, we used these methods to begin an analysis of the roles of E1--E4 and E5 in HPV pathogenesis by mutating the major splice at nt 3295. Mutation of SPA3295 did not significantly alter the ability of HPV-31 genomes to replicate transiently in keratinocytes, nor did the mutation affect the immortalization potential of HPV-31. However, genomes carrying the SPA3295 mutation were not stably maintained as viral episomes, and the resulting immortalized keratinocyte cell line contained multiple, integrated copies of the mutated HPV-31 DNA. Northern analysis indicated that cell lines immortalized with the mutant HPV-31 expressed transcripts which were similar in size and abundance to wild-type messages, including those transcripts which rely on utilization of SPA3295. RNase protection and reverse transcription-PCR revealed that mutation of SPA3295 resulted in the utilization of a cryptic splice acceptor at nt 3298. These data suggest that the requirements for stable maintenance of HPV genomes are more stringent than those for transient replication and that factors which define these requirement rely on the major splice acceptor at nt 3295. PMID- 9343170 TI - Characteristics of the Pro225His mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase that appears under selective pressure of dose escalating quinoxaline treatment of HIV-1. AB - Treatment of human immunodeficiency virus type 1 (HIV-1)-infected CEM cell cultures with escalating concentrations of the quinoxaline S-2720 resulted in an ordered appearance of single and multiple mutant virus strains that gradually became resistant to the quinoxaline and other nonnucleoside reverse transcriptase (RT) inhibitors (NNRTIs). A novel mutation, Pro225His, consistently appeared in a Val106Ala RT-mutated genetic background. The contribution of this mutation to the resistance of the mutant HIV-1 RT to NNRTIs was additive to the resistance caused by the Val106Ala mutation. Interestingly, site-directed mutagenesis studies revealed that the Pro225His-mutated RT had acquired markedly greater sensitivity to bis(heteroaryl)piperazine (BHAP U-90152) (delavirdine) but not to any of the other NNRTIs. The kinetics of inhibition of the Pro225His mutant RT by the NNRTIs (including BHAP U-90152) was not substantially different from that observed for the wild-type RT. The hypersensitivity of the mutant enzyme and virus to BHAP U 90152 could be rationally explained by the molecular-structural determinants of the RT-BHAP complex, which has recently been resolved by X-ray crystallography. PMID- 9343168 TI - Identification of domains within the human cytomegalovirus major immediate-early 86-kilodalton protein and the retinoblastoma protein required for physical and functional interaction with each other. AB - The human cytomegalovirus major immediate-early 86-kDa protein (IE2 86) plays an important role in the trans activation and regulation of HCMV gene expression. Previously, we demonstrated that IE2 86 contains three regions (amino acids [aa] 86 to 135, 136 to 290, and 291 to 364) that can independently bind to in vitro translated Rb when IE2 86 is produced as a glutathione S-transferase fusion protein (M. H. Sommer, A. L. Scully, and D. H. Spector, J. Virol. 68:6223-6231, 1994). In this report, we have elucidated the regions of Rb involved in binding to IE2 86 and have further analyzed the functional nature of the interaction between these two proteins. We find that two domains on Rb, the A/B pocket and the carboxy terminus, can each independently form a complex with IE2 86. In functional assays, we demonstrate that IE2 86 and another IE protein, IE1 72, can counter the enlarged flat cell phenotype, but not the G1/S block, which results from expression of wild-type Rb in the human osteosarcoma cell line Saos-2. Mutational analysis reveals that there are two domains on IE2 86 that can independently affect Rb function. One region (aa 241 to 369) includes the major Rb-binding domain, while the second maps to the amino-terminal region (aa 1 to 85) common to both IE2 86 and IE1 72. These data show that Rb and IE2 86 physically and functionally interact with each other via at least two separate domains and provide further support for the hypothesis that IE2 86 may exert its pleiotropic effects through the formation of multimeric protein complexes. PMID- 9343171 TI - Collectin-mediated antiviral host defense of the lung: evidence from influenza virus infection of mice. AB - Collagenous lectins (collectins) present in mammalian serum and pulmonary fluids bind to influenza virus and display antiviral activity in vitro, but their role in vivo has yet to be determined. We have used early and late isolates of H3N2 subtype influenza viruses that differ in their degree of glycosylation to examine the relationship between sensitivity to murine serum and pulmonary lectins in vitro and the ability of a virus to replicate in the respiratory tract of mice. A marked inverse correlation was found between these two parameters. Early H3 isolates (1968 to 1972) bear 7 potential glycosylation sites on hemagglutinin (HA), whereas later strains carry 9 or 10. Late isolates were shown to be much more sensitive than early strains to neutralization by the mouse serum mannose binding lectin (MBL) and rat lung surfactant protein D (SP-D) and bound greater levels of these lectins in enzyme-linked immunosorbent assays and Western blot analyses. They also replicated very poorly in mouse lungs compared to the earlier strains. Growth in the lungs was greatly enhanced, however, if saccharide inhibitors of the collectins were included in the virus inoculum. The level of SP D in bronchoalveolar lavage fluids increased on influenza virus infection. MBL was absent from lavage fluids of normal mice but could be detected in fluids from mice 3 days after infection with the virulent strain A/PR/8/34 (H1N1). The results implicate SP-D and possibly MBL as important components of the innate defense of the respiratory tract against influenza virus and indicate that the degree or pattern of glycosylation of a virus can be an important factor in its virulence. PMID- 9343172 TI - Single point mutations may affect the serotype reactivity of serotype G11 porcine rotavirus strains: a widening spectrum? AB - A panel of single and double neutralization-resistant escape mutants of serotype G11 porcine rotavirus strains A253 and YM, selected with G11 monotype- and serotype-specific neutralizing monoclonal antibodies (MAbs) to VP7, was tested in neutralization assays with hyperimmune sera raised against rotavirus strains of different serotypes. Escape mutants with an amino acid substitution in antigenic region A (amino acids [aa] 87 to 101) resulting in a residue identical or chemically similar to those present at the same positions in serotype G3 strains, at positions 87 for strain A253 and 96 for strain YM, were significantly more sensitive than the parental strains to neutralization with sera against some serotype G3 strains. Also, one YM antigenic variant (YM-5E6.1) acquired reactivity by enzyme-linked immunosorbent assay with MAbs 159, 57/8, and YO-1E2, which react with G3 strains, but not with the serotype G11 parental strain YM. Cross-adsorption studies suggested that the observed cross-neutralization by the G3-specific sera was due to the sera containing antibodies reactive with the parental strain plus antibodies reactive with the epitope(s) on the antigenic variant that mimick the serotype G3 specific one(s). Moreover, antibodies reactive with antigenic region F (aa 235 to 242) of VP7 might also be involved since cross-reactivity to serotype G3 was decreased in double mutants carrying an additional mutation, which creates a potential glycosylation site at position 238. Thus, single point mutations can affect the serotype reactivity of G11 porcine rotavirus strains with both monoclonal and polyclonal antibodies and may explain the origin of rotavirus strains with dual serotype specificity based on sequence divergence of VP7. PMID- 9343173 TI - Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins. AB - Alteration of the natural tropism of adenovirus (Ad) will permit gene transfer into specific cell types and thereby greatly broaden the scope of target diseases that can be treated by using Ad. We have constructed two Ad vectors which contain modifications to the Ad fiber coat protein that redirect virus binding to either alpha(v) integrin [AdZ.F(RGD)] or heparan sulfate [AdZ.F(pK7)] cellular receptors. These vectors were constructed by a novel method involving E4 rescue of an E4-deficient Ad with a transfer vector containing both the E4 region and the modified fiber gene. AdZ.F(RGD) increased gene delivery to endothelial and smooth muscle cells expressing alpha(v) integrins. Likewise, AdZ.F(pK7) increased transduction 5- to 500-fold in multiple cell types lacking high levels of Ad fiber receptor, including macrophage, endothelial, smooth muscle, fibroblast, and T cells. In addition, AdZ.F(pK7) significantly increased gene transfer in vivo to vascular smooth muscle cells of the porcine iliac artery following balloon angioplasty. These vectors may therefore be useful in gene therapy for vascular restenosis or for targeting endothelial cells in tumors. Although binding to the fiber receptor still occurs with these vectors, they demonstrate the feasibility of tissue-specific receptor targeting in cells which express low levels of Ad fiber receptor. PMID- 9343174 TI - Resistance to a drug blocking human immunodeficiency virus type 1 entry (RPR103611) is conferred by mutations in gp41. AB - A triterpene derived from betulinic acid (RPR103611) blocks human immunodeficiency virus type 1 (HIV-1) infection and fusion of CD4+ cells with cells expressing HIV-1 envelope proteins (gp120 and gp41), suggesting an effect on virus entry. This compound did not block infection by a subtype D HIV-1 strain (NDK) or cell-cell fusion mediated by the NDK envelope proteins. The genetic basis of drug resistance was therefore addressed by testing envelope chimeras derived from NDK and a drug-sensitive HIV-1 strain (LAI, subtype B). A drug resistant phenotype was observed for all chimeras bearing the ectodomain of NDK gp41, while the origins of gp120 and of the membrane anchor and cytoplasmic domains of gp41 had no apparent role. The envelope gene of a LAI variant, fully resistant to the antiviral effect of RPR103611, was cloned and sequenced. Its product differed from the parental sequence at two positions in gp41, with changes of arginine 22 to alanine (R22A) and isoleucine 84 to serine (I84S), the gp120 being identical. In the context of LAI gp41, the I84S substitution was sufficient for drug resistance. Therefore, in two different systems, differences in gp41 were associated with sensitivity or resistance to RPR103611. Modifications of gp41 can affect the quaternary structure of gp120 and gp41 and the accessibility of gp120 to antiviral agents such as neutralizing antibodies. However, a direct effect of RPR103611 on a gp41 target must also be envisioned, in agreement with the blocking of apparently late steps of HIV-1 entry. This compound could be a valuable tool for structure-function studies of gp41. PMID- 9343175 TI - Usage of the coreceptors CCR-5, CCR-3, and CXCR-4 by primary and cell line adapted human immunodeficiency virus type 2. AB - The chemokine receptors CCR-5 and CXCR-4, and possibly CCR-3, are the principal human immunodeficiency virus type 1 (HIV-1) coreceptors, apparently interacting with HIV-1 envelope, in association with CD4. Cell lines coexpressing CD4 and these chemokine receptors were infected with a panel of seven primary HIV-2 isolates passaged in peripheral blood mononuclear cells (PBMC) and three laboratory HIV-2 strains passaged in T-cell lines. The CCR-5, CCR-3, and CXCR-4 coreceptors could all be used by HIV-2. The ability to use CXCR-4 represents a major difference between HIV-2 and the closely related simian immunodeficiency viruses. Most HIV-2 strains using CCR-5 could also use CCR-3, sometimes with similar efficiencies. As observed for HIV-1, the usage of CCR-5 or CCR-3 was observed principally for HIV-2 strains derived from asymptomatic individuals, while HIV-2 strains derived from AIDS patients used CXCR-4. However, there were several exceptions, and the patterns of coreceptor usage seemed more complex for HIV-2 than for HIV-1. The two T-tropic HIV-2 strains tested used CXCR-4 and not CCR-5, while T-tropic HIV-1 can generally use both. Moreover, among five primary HIV-2 strains all unable to use CXCR-4, three could replicate in CCR-5-negative PBMC, which has not been reported for HIV-1. These observations suggest that the CCR-5 coreceptor is less important for HIV-2 than for HIV-1 and indicate that HIV 2 can use other cell entry pathways and probably other coreceptors. One HIV-2 isolate replicating in normal or CCR-5-negative PBMC failed to infect CXCR-4+ cells or the U87MG-CD4 and sMAGI cell lines, which are permissive to infection by HIV-2 but not by HIV-1. This suggests the existence of several HIV-2-specific coreceptors, which are differentially expressed in cell lines and PBMC. PMID- 9343177 TI - Antibody-induced and cytoskeleton-mediated redistribution and shedding of viral glycoproteins, expressed on pseudorabies virus-infected cells. AB - Fluorescein isothiocyanate-labeled porcine pseudorabies virus (PrV) polyclonal antibodies were added to PrV-infected swine kidney cells in vitro at 37 degrees C. In approximately 47% of the infected cells, the addition induced passive patching and subsequent energy- and microtubule-dependent capping of all viral envelope glycoproteins, expressed on the plasma membranes of the infected cells. Further contraction and extrusion of the capped viral glycoproteins occurred in approximately 30% of the capped cells 2 h after the addition of antibodies and was accompanied by a concentration of F-actin beneath the caps. At that time, about 18% of the extruded caps were shed spontaneously into the surrounding medium. Mechanical force released 85% of the extruded caps, leaving viable cells with no microscopically detectable levels of viral glycoproteins on their plasma membranes. Experiments with PrV deletion mutants showed that viral glycoproteins gE and gI are important in triggering viral glycoprotein redistribution. Since the PrV gE-gI complex exhibits Fc receptor activity which facilitates capping, the importance of gE and gI may be partially explained by antibody bipolar bridging. PMID- 9343178 TI - Adeno-associated virus type 2-mediated transduction in primary human bone marrow derived CD34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. AB - Although the adeno-associated virus type 2 (AAV) is known to possess a broad host range that transcends the species barrier, we suggested in an earlier study that AAV infection of human cells is receptor mediated (S. Ponnazhagan et al., J. Gen. Virol. 77:1111-1122, 1996). In the present studies, we investigated the ability of AAV to infect primary human hematopoietic progenitor cells capable of multilineage differentiation. Bone marrow-derived CD34+ cells from 12 hematologically normal volunteer donors were infected with a recombinant AAV containing the beta-galactosidase gene under the control of the cytomegalovirus immediate-early promoter (vCMVp-lacZ). Whereas 15 to 80% of the cells from approximately 50% of the donors showed various levels of lacZ gene expression, the expression was undetectable in cells from the remaining donors. However, if cells from both sets of donors were stimulated with various combinations of cytokines to induce differentiation into myeloid and lymphoid lineages following AAV infection, then the level of expression of the transduced gene increased up to 20-fold over a period of 14 days. The results of virus-binding assays suggested that the observed difference between the two groups was due to the differential susceptibility of CD34+ cells to AAV infection rather than to differences in transcription and translation of the transduced gene. To corroborate these results, CD34+ cells from the two donor groups, KB (human nasopharyngeal carcinoma) cells, and M07e (human megakaryocytic leukemia) cells were infected with vCMVp-lacZ. KB cells served as a positive control for AAV infection, and M07e cells served as a negative control. Whereas abundant hybridization to the single-stranded viral DNA on Southern blots was detected in KB and CD34+ cells that were positive for lacZ gene expression, little activity was detected in M07e and CD34+ cells that did not show expression of the lacZ gene. These results suggest that the levels of expression of the putative cellular receptor for AAV vary widely in CD34+ cells from different donors. These studies have implications for the potential use of AAV vectors in human gene therapy involving primary human primitive hematopoietic stem and progenitor cells. PMID- 9343176 TI - High viral burden and rapid CD4+ cell depletion in human immunodeficiency virus type 1-infected SCID-hu mice suggest direct viral killing of thymocytes in vivo. AB - The mechanism of CD4+ cell loss in lymphoid organs is unknown. In this study, human immunodeficiency virus (HIV) infection of human fetal thymus/liver implants in severe combined immunodeficient mice was used to investigate the mechanism of HIV-induced depletion of CD4-bearing cells in vivo. The implants were assessed for depletion of CD4+ thymocytes, apoptosis, and viral burden. We detected two phases of CD4 cell depletion, an initial rapid phase and a more gradual later phase. Compared to mock-infected implants, HIV-infected implants did not demonstrate detectable increases in the levels of apoptosis while severe depletion of CD4-bearing cells was ongoing. During peak loss of CD4+ cells, high viral burden was observed, suggesting that loss of CD4+ cells in this in vivo system is due to direct killing of infected thymocytes. Increased levels of apoptosis were observed during the later phase of thymocyte depletion; however, these apoptotic cells lacked CD4. This finding suggests that a second indirect mechanism may be responsible for the destruction of CD4- CD8+ thymocytes in vivo. Taken together, these results suggest that CD4+ and CD4- cells may die by different mechanism(s). PMID- 9343179 TI - Rotavirus is released from the apical surface of cultured human intestinal cells through nonconventional vesicular transport that bypasses the Golgi apparatus. AB - Rotaviruses are nonenveloped viruses that infect enterocytes of the small intestine and cause severe infantile gastroenteritis. It was previously thought that rotavirus exits cells by lysis, but this behavior does not match the local pathogenesis of the virus. In this study, we have investigated the release of the simian rotavirus strain (RRV) from the polarized intestinal Caco-2 cells. We found that RRV is released almost exclusively from the apical pole of Caco-2 cells before any cells lyse. Using confocal laser scanning microscopy and drugs that inhibit vesicular transport, we studied the RRV transport route from the endoplasmic reticulum (ER) to the apical side of intestinal cells. We demonstrated that RRV exits from the ER through a carbonyl cyanide m chlorophenylhydrazone-sensitive vesicular transport. RRV staining was never found within the Golgi apparatus or lysosomes, suggesting that the RRV intracellular pathway does not involve these organelles. This finding was confirmed by treatment with monensin or NH4Cl, which do not affect release of RRV. Electron microscopic analysis revealed RRV containing small smooth vesicles in the apical area and free virions outside the cell in the brush border, consistent with a vesicular vectorial transport of virus. These results may provide, for the first time, a cellular explanation of the pathogenesis of rotavirus. PMID- 9343180 TI - Mutational analysis of the role of glycoprotein I in varicella-zoster virus replication and its effects on glycoprotein E conformation and trafficking. AB - The contributions of the glycoproteins gI (ORF67) and gE (ORF68) to varicella zoster virus (VZV) replication were investigated in deletion mutants made by using cosmids with VZV DNA derived from the Oka strain. Deletion of both gI and gE prevented virus replication. Complete deletion of gI or deletions of 60% of the N terminus or 40% of the C terminus of gI resulted in a small plaque phenotype as well as reduced yields of infectious virus. Melanoma cells infected with gI deletion mutants formed abnormal polykaryocytes with a disrupted organization of nuclei. In the absence of intact gI, gE became localized in patches on the cell membrane, as demonstrated by confocal microscopy. A truncated N-terminal form of gI was transported to the cell surface, but its expression did not restore plaque morphology or infectivity. The fusogenic function of gH did not compensate for gI deletion or the associated disruption of the gE-gI complex. These experiments demonstrated that gI was dispensable for VZV replication in vitro, whereas gE appeared to be required. Although VZV gI was dispensable, its deletion or mutation resulted in a significant decrease in infectious virus yields, disrupted syncytium formation, and altered the conformation and distribution of gE in infected cells. Normal cell-to-cell spread and replication kinetics were restored when gI was expressed from a nonnative locus in the VZV genome. The expression of intact gI, the ORF67 gene product, is required for efficient membrane fusion during VZV replication. PMID- 9343181 TI - Neutralizing antibodies against the V3 loop of human immunodeficiency virus type 1 gp120 block the CD4-dependent and -independent binding of virus to cells. AB - The CD4 molecule is an essential receptor for human immunodeficiency virus type 1 (HIV-1) through high-affinity interactions with the viral external envelope glycoprotein gp120. Previously, neutralizing monoclonal antibodies (MAbs) specific to the third hypervariable domain of gp120 (the V3 loop) have been thought to block HIV infection without affecting the binding of HIV particles to CD4-expressing human cells. However, here we demonstrate that this conclusion was not correct and was due to the use of soluble gp120 instead of HIV particles. Indeed, neutralizing anti-V3 loop MAbs inhibited completely the binding and entry of HIV particles into CD4+ human cells. In contrast, the binding of virus was only partially inhibited by neutralizing anti-CD4 MAbs against the gp120 binding site in CD4, which, like the anti-V3 loop MAbs, completely inhibited HIV entry and infection. Nonneutralizing control MAbs against either the V3 loop or the N or C terminus of gp120 had no significant effect on HIV binding and entry. HIV-1 particles were also found to bind human and murine cells expressing or not expressing the human CD4 molecule. Interestingly, the binding of HIV to CD4+ murine cells was inhibited by both anti-V3 and anti-CD4 MAbs, whereas the binding to human and murine CD4- cells was affected only by anti-V3 loop MAbs. The effect of anti-V3 loop neutralizing MAbs on the HIV binding to cells appears not to be the direct consequence of gp120 shedding from HIV particles or of a decreased affinity of CD4 or gp120 for binding to its surface counterpart. Taken together, our results suggest the existence of CD4-dependent and -independent binding events involved in the attachment of HIV particles to cells; in both of these events, the V3 loop plays a critical role. As murine cells lack the specific cofactor CXCR4 for HIV-1 entry, other cell surface molecules besides CD4 might be implicated in stable binding of HIV particles to cells. PMID- 9343182 TI - The adenovirus E3-10.4K/14.5K complex mediates loss of cell surface Fas (CD95) and resistance to Fas-induced apoptosis. AB - Cytotoxic T cells use Fas (CD95), a member of the tumor necrosis factor (TNF) receptor superfamily, to eliminate virus-infected cells by activation of the apoptotic pathway for cell death. The adenovirus E3 region encodes several proteins that modify immune defenses, including TNF-dependent cell death, which may allow this virus to establish a persistent infection. Here we show that, as an early event during infection, the adenovirus E3-10.4K/14.5K complex selectively induces loss of Fas surface expression and blocks Fas-induced apoptosis of virus-infected cells. Loss of surface Fas occurs within the first 4 h postinfection and is not due to decreased production of Fas protein. The decrease in surface Fas is distinct from the 10.4K/14.5K-mediated loss of the epidermal growth factor receptor on the same cells, because intracellular stores of Fas are not affected. Further, 10.4K/14.5K, which was previously shown to protect against TNF cytolysis, does not induce a loss of TNF receptor, indicating that this complex mediates more than one function to block host defense mechanisms. These results suggest yet another mechanism by which adenovirus modulates host cytotoxic responses that may contribute to persistent infection by human adenoviruses. PMID- 9343183 TI - The null mutant of the U(L)31 gene of herpes simplex virus 1: construction and phenotype in infected cells. AB - Earlier studies have shown that the U(L)31 protein is homogeneously distributed throughout the nucleus and cofractionates with nuclear matrix. We report the construction from an appropriate cosmid library a deletion mutant which replicates in rabbit skin cells carrying the U(L)31 gene under a late (gamma1) viral promoter. The mutant virus exhibits cytopathic effects and yields 0.01 to 0.1% of the yield of wild-type parent virus in noncomplementing cells but amounts of virus 10- to 1,000-fold higher than those recovered from the same cells 3 h after infection. Electron microscopic studies indicate the presence of small numbers of full capsids but a lack of enveloped virions. Viral DNA extracted from the cytoplasm of infected cells exhibits free termini indicating cleavage/packaging of viral DNA from concatemers for packaging into virions, but analyses of viral DNAs by pulsed-field electrophoresis indicate that at 16 h after infection, both the yields of viral DNA and cleavage of viral DNA for packaging are decreased. The repaired virus cannot be differentiated from the wild-type parent. These results suggest the possibility that U(L)31 protein forms a network to enable the anchorage of viral products for the synthesis and/or packaging of viral DNA into virions. PMID- 9343184 TI - A century of tobamovirus evolution in an Australian population of Nicotiana glauca. AB - The evolution over the past century of two tobamoviruses infecting populations of the immigrant plant Nicotiana glauca in New South Wales (NSW), Australia, has been studied. This plant species probably entered Australia in the 1870s. Isolates of the viruses were obtained from N. glauca specimens deposited in the NSW Herbarium between 1899 and 1972, and others were obtained from living plants in 1985 and 1993. It was found that the NSW N. glauca population was infected with tobacco mosaic tobamovirus (TMV) and tobacco mild green mosaic tobamovirus (TMGMV) before 1950 but only with TMGMV after that date. Half the pre-1950 infections were mixtures of the two viruses, and one was a recombinant. Remarkably, sequence analyses showed no increase in the genetic diversity among the TMGMV isolates over the period. However, for TMV, the genetic diversity of synonymous (but not of nonsynonymous) differences between isolates varied and was correlated with their time of isolation. TMV accumulated to smaller concentrations than TMGMV in N. glauca plants, and in mixed experimental infections, the accumulation of TMV, but not of TMGMV, was around 1/10 that in single infections. However, no evidence was found of isolate-specific interaction between the viruses. We conclude that although TMV may have colonized N. glauca in NSW earlier or faster than TMGMV, the latter virus caused a decrease of the TMV population below a threshold at which deleterious mutations were eliminated. This phenomenon, called Muller's ratchet, or a "mutational meltdown," probably caused the disappearance of TMV from the niche. PMID- 9343185 TI - Readthrough activation of early adenovirus E1b gene transcription. AB - In cells productively infected with adenovirus type 5, transcription is not terminated between the E1a gene and the adjacent downstream E1b gene. Insertion of the mouse beta(maj)-globin transcription termination sequence (GGT) into the E1a coding region dramatically reduces early, but not late, E1b expression (E. Falck-Pedersen, J. Logan, T. Shenk, and J. E. Darnell, Jr., Cell 40:897-905, 1985). In the study described herein, we showed that base substitution mutations in the globin DNA that specifically relieved transcription termination also restored early E1b promoter activity in cis, establishing that maximal early E1b expression requires readthrough transcription originating from the adjacent upstream gene. To identify potential targets of readthrough activation, a series of recombinant viruses with double mutations was constructed. Each double-mutant virus strain had the transcription termination sequences in the first exon of E1a and a deletion within the transcription control region of E1b. Early E1b expression from the double-mutant strains was more defective than that from strains containing either mutation alone, indicating that the deleted regions (positions -362 to -35) are not the target for readthrough activation. Two findings suggested that a cis-dominant property of early viral templates is important for readthrough activation. First, the early E1b defect caused by the GGT insertion was not complemented in trans by factors present in late-infected cells. Second, restoration of E1b transcription at late times occurred concurrently with viral DNA replication. Readthrough activation may help convert virion DNA into a transcriptionally competent template prior to DNA replication and late transcription. PMID- 9343187 TI - Evolutionary dynamics of genetic variation in Epstein-Barr virus isolates of diverse geographical origins: evidence for immune pressure-independent genetic drift. AB - The question whether immune pressure exerted by cytotoxic T lymphocytes (CTLs) can influence the long-term evolution of genetically stable viruses such as Epstein-Barr virus (EBV) has generated considerable scientific interest, primarily due to its important implications for the overall biology of the virus. While arguing for a role of CTLs in the evolution of viruses, it is important to differentiate between genetic variation in virus and immune recognition of these variant virus by CTLs. To assess the role of genetic selection in the long-term evolution of EBV, we have analyzed a large panel of type 1 EBV isolates from African, Southeast Asian, Papua-New Guinean (PNG), and Australian Caucasian individuals. Seven different regions of the EBV genome, which include nine CTL epitopes restricted through a range of HLA class I alleles, were sequenced and compared. Although numerous nucleotide changes were identified within these isolates, comparison of synonymous and nonsynonymous substitutions in the CTL epitope indicated that the genetic variation was generated mostly independently of immune selection pressure. Surprisingly, an inverse correlation between genetic variation within certain CTL epitopes and the frequency distribution of HLA alleles that present the CTL epitopes was seen, suggesting that the evolutionary pressures on the CTL epitopes of the virus may be toward their conservation rather than their inactivation. Furthermore, molecular evolutionary genetic analysis of nucleotide sequences revealed that viral isolates from PNG are evolving as a lineage distinct from isolates from African, Southeast Asian, and Australian Caucasian individuals. PMID- 9343186 TI - Functional involvement of polypyrimidine tract-binding protein in translation initiation complexes with the internal ribosome entry site of foot-and-mouth disease virus. AB - The synthesis of picornavirus polyproteins is initiated cap independently far downstream from the 5' end of the viral RNA at the internal ribosome entry site (IRES). The cellular polypyrimidine tract-binding protein (PTB) binds to the IRES of foot-and-mouth disease virus (FMDV). In this study, we demonstrate that PTB is a component of 48S and 80S ribosomal initiation complexes formed with FMDV IRES RNA. The incorporation of PTB into these initiation complexes is dependent on the entry of the IRES RNA, since PTB and IRES RNA can be enriched in parallel either in 48S or 80S ribosomal complexes by stage-specific inhibitors of translation initiation. The formation of the ribosomal initiation complexes with the IRES occurs slowly, is temperature dependent, and correlates with the incorporation of PTB into these complexes. In a first step, PTB binds to the IRES, and then the small ribosomal subunit encounters this PTB-IRES complex. Mutations in the major PTB-binding site interfere simultaneously with the formation of initiation complexes, translation efficiency, and PTB cross-linking. PTB stimulates translation directed by the FMDV IRES in a rabbit reticulocyte lysate depleted of internal PTB, and the efficiency of translation can be restored to the original level by the addition of PTB. These results indicate that PTB plays an important role in the formation of initiation complexes with FMDV IRES RNA and in stimulation of internal translation initiation with this picornavirus. PMID- 9343188 TI - In vitro polymerase activity of Thogoto virus: evidence for a unique cap snatching mechanism in a tick-borne orthomyxovirus. AB - The tick-borne Thogoto virus (THOV) is the type species of a new genus in the family Orthomyxoviridae. Its genome comprises six segments of single-stranded, negative-sense RNA. Each segment possesses conserved regions of semicomplementary nucleotides at the 3' and 5' termini which strongly resemble those of influenza virus. An in vitro polymerase assay based on reconstituted THOV viral cores was developed, and activity was shown to rely on an interaction between the conserved 3'- and 5'-terminal sequences and to be primer dependent. Addition of globin mRNA primed transcription, catalyzing the addition of an extra nucleotide to the transcripts, corresponding to the 5'-terminal m7G cap residue. Priming with various cap analogs suggested that THOV transcription is initiated preferentially with m7GpppAm and involves base pairing. This is the first experimental evidence of endonuclease activity in THOV as part of a unique cap-snatching mechanism. PMID- 9343189 TI - Striking conformational similarities between the transcription promoters of Thogoto and influenza A viruses: evidence for intrastrand base pairing in the 5' promoter arm. AB - In the accompanying report, we describe an in vitro polymerase assay based on reconstituted Thogoto virus (THOV) cores which provided evidence of a double stranded vRNA promoter consisting of both the 3' and 5' sequences of vRNA (M. B. Leahy, J. T. Dessens, and P. A. Nuttall, J. Virol. 71:8347-8351, 1997). This system was used to investigate further the THOV vRNA promoter structure by using short, synthetic vRNA promoters. The results obtained show that interstrand base pairing between residues 10 and 11 of the 3' promoter arm with residues 11 and 12 of the 5' promoter arm, respectively, is important for promoter activity. In addition, intrastrand base pairing between residues 2 and 3 with residues 9 and 8 of the 5' promoter arm, respectively, was shown to be involved in promoter activity, while no evidence of intrastrand base pairing between residues 2 and 9 of the 3' promoter arm was obtained. These observations are consistent with a hook-like structure in the 5' promoter arm of the THOV promoter. The THOV cores were able to transcribe an influenza A virus (FLUA) vRNA-like promoter, as well as hybrid THOV-FLUA promoters. Hence, the THOV and FLUA vRNA promoters appear to be both structurally and functionally similar. PMID- 9343190 TI - Arginine-glycine-aspartic acid-specific binding by foot-and-mouth disease viruses to the purified integrin alpha(v)beta3 in vitro. AB - The integrin alpha(v)beta3 has been shown to act as the receptor for internalization of foot-and-mouth disease virus (FMDV) (A12), with attachment being through a highly conserved RGD motif located on the G-H loop of viral capsid protein VP1. In addition, however, we have recently shown that efficient infection of culture-grown cells by FMDV (O1BFS) requires binding to cell surface heparan sulfate. In this study, we have used a solid-phase receptor binding assay to characterize the binding by FMDV to purified alpha(v)beta3 in the absence of heparan sulfate and other cell surface components. In this assay, FMDV (O1BFS) successfully replicated authentic ligand binding by cellular alpha(v)beta3 in terms of its high affinity, dependence on divalent cations, and activation by manganese ions. Virus binding to this preparation of alpha(v)beta3 was exquisitely sensitive to competition by short RGD-containing peptides (50% inhibition at < 10(-8) M peptide), and this inhibition was highly sequence specific, with the equivalent RGE peptide being at least 10(4) fold less effective as a competitor. Representative viruses of the other six serotypes of FMDV bound to alpha(v)beta3 in a similar RGD-specific manner, although significant differences in sensitivity to RGD peptides suggest that the affinity of the different FMDV serotypes for alpha(v)beta3 is influenced, in part, by the variable amino acid residues in the VP1 G-H loop on either side of the RGD. PMID- 9343191 TI - Investigation of the attenuation exhibited by a molecularly cloned chicken anemia virus isolate by utilizing a chimeric virus approach. AB - Molecular cloning of the Cux-1 isolate of chicken anemia virus (CAV), which had been passaged 173 times in cell culture, resulted in the isolation of an attenuated strain, designated cloned isolate 10, which reverted to virulence following 10 passages in young chicks (D. Todd, T. J. Connor, V. M. Calvert, J. L. Creelan, B. M. Meehan, and M. S. McNulty, Avian Pathol. 24:171-187, 1995). The attenuated cloned isolate 10 differs from the molecularly cloned pathogenic Cux-1 isolate in that it possesses a 21-nucleotide insertion within the nontranscribed region of the CAV genome and 17 individual nucleotide substitutions dispersed throughout the genome. Comparative analyses with other published CAV sequences indicated that cloned isolate 10 was unique at nine nucleotide positions and at five amino acid positions. The molecular basis of the attenuation exhibited by cloned isolate 10 was investigated by evaluating the pathogenicities of two sets of complementary chimeric viruses. These sets were produced by transfection with chimeric double-stranded replicative-form (RF) DNA equivalents that contained DNA sequences derived from cloned isolate 10 and the pathogenic cloned Cux-1 isolate. The construction of the chimeric RFs exploited the occurrence of unique EcoRI, PstI, and BamHI restriction sites, which allowed their respective circular CAV RFs to be manipulated as three restriction fragments of 0.58, 0.93, and 0.71 kbp. Examination of the levels of anemia and gross pathology in the thymuses and bone marrows of 14 day-old specific-pathogen-free chicks following infection of 1-day old chicks with the chimeric and cloned parental isolates indicated that nucleotide changes in each of the three genomic regions contributed towards attenuation. The significance of this result to the development and use of live attenuated CAV vaccines is discussed. PMID- 9343193 TI - Regulation of receptor binding affinity of influenza virus hemagglutinin by its carbohydrate moiety. AB - The hemagglutinin (HA) of the fowl plague virus (FPV) strain of influenza A virus has two N-linked oligosaccharides attached to Asn123 and Asn149 in the vicinity of the receptor binding site. The effect of these carbohydrate side chains on the binding of HA to neuraminic acid-containing receptors has been analyzed. When the oligosaccharides were deleted by site-specific mutagenesis, HA expressed from a simian virus 40 vector showed enhanced hemadsorbing activity. Binding was so strong under these conditions that erythrocytes were no longer released by viral neuraminidase and that release was significantly reduced when neuraminidase from Vibrio cholerae was used. Similarly, when these oligosaccharides were removed selectively from purified viruses by N-glycosidase F, such virions were unable to elute from receptors, although they retained neuraminidase activity. Thus, release of FPV from cell receptors depends on the presence of the HA glycans at Asn123 and Asn149. On the other hand, receptor binding was abolished when these oligosaccharides were sialylated after expression in the absence of neuraminidase (M. Ohuchi, A. Feldmann, R. Ohuchi, and H.-D. Klenk, Virology 212:77-83, 1995). These observations indicate that the receptor affinity of FPV HA is controlled by oligosaccharides adjacent to the receptor binding site. PMID- 9343192 TI - Studies of AIDS vaccination using an ex vivo feline immunodeficiency virus model: protection conferred by a fixed-cell vaccine against cell-free and cell associated challenge differs in duration and is not easily boosted. AB - Cats immunized with cells infected with a primary isolate of feline immunodeficiency virus (FIV) and fixed with paraformaldehyde were challenged with cell-free or cell-associated homologous virus obtained ex vivo. Complete protection was observed in animals challenged with cell-free virus 4 months after completion of vaccination (p.v.) or with cell-associated virus 12 months p.v. In contrast, no protection was observed in cats challenged with cell-free virus 12 or 28 months p.v. or with cell-associated virus 37.5 months p.v. Prior to the 28- and 37.5-month challenges, the animals had received a booster dose of vaccine that had elicited a robust anamnestic immune response. These results show that vaccine-induced protection against ex vivo FIV is achievable but is relatively short-lived and can be difficult to boost. PMID- 9343194 TI - Mutations in coat protein binding sites of alfalfa mosaic virus RNA 3 affect subgenomic RNA 4 accumulation and encapsidation of viral RNAs. AB - The 3'-untranslated regions (3'-UTRs) of the three RNAs of alfalfa mosaic virus (AMV) contain a specific binding site for coat protein (CP) and act as a promoter for minus-strand RNA synthesis by the purified AMV RNA-dependent RNA polymerase (RdRp) in an in vitro assay. Binding of CP to the viral RNAs is required to initiate infection. The sequence of the 3'-terminal 39 nucleotides of AMV RNA 3 can be folded into two stem-loop structures flanked by three single-stranded AUGC sequences and represents a CP binding site. Mutations in this sequence that are known to interfere with CP binding in vitro were introduced into an infectious clone of RNA 3, and mutant RNA transcripts were used as templates in the in vitro RdRp assay and to infect protoplasts and plants. Mutation of AUGC motif 2 or disruption of the stem of the 3'-proximal hairpin 1 interfered with CP binding in vitro but not with minus-strand promoter activity in vitro or replication of RNA 3 in vivo. However, hairpin 1 appeared to be essential for encapsidation of RNA 3. Reversion of three G-C base pairs in hairpin 1 had no effect on CP binding but interfered with minus-strand promoter activity in vitro and with RNA 3 replication in vivo. It is concluded that the viral RdRp and CP recognize different elements in the 3'-UTRs of AMV RNAs. Moreover, several mutations that interfered with CP binding in vitro interfered with the accumulation in vivo of RNA 4, the subgenomic messenger for CP, but not with the accumulation of RNA 3. PMID- 9343195 TI - Virus-specific, CD8+ major histocompatibility complex class I-restricted cytotoxic T lymphocytes in lymphocytic choriomeningitis virus-infected beta2 microglobulin-deficient mice. AB - Following infection with lymphocytic choriomeningitis virus (LCMV), normal adult mice generate virus-specific, major histocompatibility complex (MHC) class I restricted cytotoxic T lymphocytes (CTL) which clear the virus after intraperitoneal infection or cause death following intracranial (i.c.) infection. We have investigated the response of beta2-microglobulin-deficient (beta2m-) mice of the H-2d haplotype (KOD mice) to LCMV infection. Unlike H-2b beta2m- mice, which generate CD4+ MHC class II-restricted CTL in response to LCMV, KOD mice generate high levels of CD8+ MHC class I-restricted, virus-specific CTL. These CTL are specific for the LCMV nucleoprotein epitope (residues 118 to 126) in association with the Ld class I molecule, analogous to the CTL response in wild type mice. KOD mice are also susceptible to lethal LCM disease, with 75 to 80% of the mice dying 7 to 9 days following i.c. infection with virus. Similar to results with normal mice, lethal LCM disease in KOD mice is prevented by in vivo depletion of CD8+ T cells prior to i.c. infection. In contrast to wild-type mice, however, KOD mice cannot control LCMV and become persistently infected. Overall, these results demonstrate that beta2m is not an absolute requirement for presentation of endogenous antigen on Ld or for induction of virus-specific Ld restricted CTL in vivo. PMID- 9343196 TI - Structure-function analysis of the gE-gI complex of feline herpesvirus: mapping of gI domains required for gE-gI interaction, intracellular transport, and cell to-cell spread. AB - Alphaherpesvirus glycoproteins gE and gI form a noncovalently associated hetero oligomeric complex, which is involved in cell-to-cell spread. In the absence of gI, feline herpesvirus (FHV) gE is transport incompetent and fully retained in the endoplasmic reticulum. Here, we assess the effect of progressive C-terminal truncations of FHV gI on the biosynthesis, intracellular transport, and function of the gE-gI complex. The truncated gI proteins were coexpressed with gE in the vaccinia virus-based vTF7-3 expression system. The results were corroborated and extended by studying FHV recombinants expressing truncated gI derivatives. The following conclusions can be drawn. (i) Deletion of the cytoplasmic tail, the transmembrane region plus the C-terminal half of the ectodomain of gI, does not affect intracellular transport of gE. Apparently, the N-terminal 166 residues of gI constitute a domain involved in gE-gI interaction. (ii) A region mediating stable association with gE is located within the N-terminal 93 residues of gI. (iii) The cytoplasmic domain of gI is not essential for gE-gI-mediated cell-to cell transmission of FHV, as judged from plaque morphology. Deletion of the cytoplasmic tail of gI reduced plaque size by only 35%. (iv) Recombinants expressing the N-terminal 166 residues of gI display a small-plaque phenotype but produce larger plaques than recombinants with a disrupted gI gene. Thus, a complex consisting of gE and the N-terminal half of the gI ectodomain may retain residual biological activity. The implications of these findings for gE-gI interaction and function are discussed. PMID- 9343197 TI - Promiscuous use of CC and CXC chemokine receptors in cell-to-cell fusion mediated by a human immunodeficiency virus type 2 envelope protein. AB - The CC chemokine receptors CCR5, CCR2, and CCR3 and the CXC chemokine receptor CXCR4 have been implicated as CD4-associated cofactors in the entry of primary and cell line-adapted human immunodeficiency virus type 1 (HIV-1) strains. CXCR4 is also a receptor for T-cell-line-adapted, CD4-independent strains of HIV-2. With the exception of this latter example, little has been reported on the entry cofactors used by HIV-2 strains. Here we show that a CD4-dependent, T-cell-line adapted HIV-2 strain uses CXCR4 and, to a lesser extent, CCR3 for fusion with and infectious entry into cells. In a cell-to-cell fusion assay, the envelope protein of this virus can utilize a wider repertoire of chemokine receptors to induce fusion. These include CCR1, CCR2, CCR3, CCR4, CCR5, CXCR2, and CXCR4. Kinetic analysis indicated that cell lines expressing the receptors that support infection, CXCR4 and CCR3, form syncytia more rapidly than do cell lines expressing the other receptors. Nevertheless, although less efficient, fusion with CXCR2 expressing cells was specific, since it was inhibited by antibodies against CXCR2. The extensive use of chemokine receptors in cell-to-cell fusion has implications for understanding the molecular basis of CD4-chemokine receptor induced lentivirus fusion and may have relevance for syncytium formation and the direct cell-to-cell transfer of virus in vivo. PMID- 9343198 TI - Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity. AB - The NS5B protein of the hepatitis C virus (HCV) is an RNA-dependent RNA polymerase (RdRp) (S.-E. Behrens, L. Tomei, and R. De Francesco, EMBO J. 15:12 22, 1996) that is assumed to be required for replication of the viral genome. To further study the biochemical and structural properties of this enzyme, an NS5B hexahistidine fusion protein was expressed with recombinant baculoviruses in insect cells and purified to near homogeneity. The enzyme was found to have a primer-dependent RdRp activity that was able to copy a complete in vitro transcribed HCV genome in the absence of additional viral or cellular factors. Filter binding assays and competition experiments showed that the purified enzyme binds RNA with no clear preference for HCV 3'-end sequences. Binding to homopolymeric RNAs was also examined, and the following order of specificity was observed: poly(U) > poly(G) > poly(A) > poly(C). An inverse order was found for the RdRp activity, which used poly(C) most efficiently as a template but was inactive on poly(U) and poly(G), suggesting that a high binding affinity between polymerase and template interferes with processivity. By using a mutational analysis, four amino acid sequence motifs crucial for RdRp activity were identified. While most substitutions of conserved residues within these motifs severely reduced the enzymatic activities, a single substitution in motif D which enhanced the RdRp activity by about 50% was found. Deletion studies indicate that amino acid residues at the very termini, in particular the amino terminus, are important for RdRp activity but not for RNA binding. Finally, we found a terminal transferase activity associated with the purified enzyme. However, this activity was also detected with NS5B proteins with an inactive RdRp, with an NS4B protein purified in the same way, and with wild-type baculovirus, suggesting that it is not an inherent activity of NS5B. PMID- 9343199 TI - Adeno-associated virus vector integration junctions. AB - Vectors derived from adeno-associated virus (AAV) have the potential to stably transduce mammalian cells by integrating into host chromosomes. Despite active research on the use of AAV vectors for gene therapy, the structure of integrated vector proviruses has not previously been analyzed at the DNA sequence level. Studies on the integration of wild-type AAV have identified a common site specific integration locus on human chromosome 19; however, most AAV vectors do not appear to integrate at this locus. To improve our understanding of AAV vector integration, we analyzed the DNA sequences of several integrated vector proviruses. HeLa cells were transduced with an AAV shuttle vector, and integrated proviruses containing flanking human DNA were recovered as bacterial plasmids for further analysis. We found that AAV vectors integrated as single-copy proviruses at random chromosomal locations and that the flanking HeLa DNA at integration sites was not homologous to AAV or the site-specific integration locus of wild type AAV. Recombination junctions were scattered throughout the vector terminal repeats with no apparent site specificity. None of the integrated vectors were fully intact. Vector proviruses with nearly intact terminal repeats were excised and amplified after infection with wild-type AAV and adenovirus. Our results suggest that AAV vectors integrate by nonhomologous recombination after partial degradation of entering vector genomes. These findings have important implications for the mechanism of AAV vector integration and the use of these vectors in human gene therapy. PMID- 9343200 TI - Control of adeno-associated virus type 2 cap gene expression: relative influence of helper virus, terminal repeats, and Rep proteins. AB - Adeno-associated virus type 2 (AAV-2) gene expression is tightly controlled by functions of the helper virus as well as by the products of its own viral rep gene. Double-immunofluorescence studies of Rep and VP protein expression in cells coinfected with AAV-2 and adenovirus type 2 showed that a large proportion of these cells expressed Rep78 and Rep52 but no capsid proteins. The percentage of Rep78/Rep52- and capsid protein-positive cells was strongly influenced by the relative ratio of AAV-2 to adenovirus type 2. In contrast, nearly all cells positive for Rep68/Rep40 were also positive for capsid protein expression. Examination of p40 promoter transactivation by individual Rep proteins in the presence of adenovirus, however, showed that both Rep78 and Rep68 efficiently stimulated p40 mRNA accumulation and capsid protein expression. This strong transactivation was reliant upon the presence of terminal repeats and correlated with template amplification. In replication-deficient expression constructs, transactivation was observed only with Rep68 and was dependent on the linear Rep binding site within the left terminal repeat which was detected in the presence of high adenovirus concentrations. In the absence of any terminal repeat sequences, Rep68 expression again led to a minor transactivation of capsid protein expression which was detectable only at low adenovirus concentrations. This low level of transactivation of capsid protein expression by Rep proteins in the absence of terminal repeats resulted in a lower efficiency of capsid assembly. The data show a dominant influence of adenovirus type 2 functions on AAV-2 gene expression, a requirement for terminal repeats for strong transactivation of the p40 promoter by Rep proteins, and differential influences of Rep78 and Rep68 on AAV-2 promoters. Implications for the production of recombinant AAV-2 vectors are discussed. PMID- 9343201 TI - Effects of mutations within and adjacent to the terminal repeats of hepatitis B virus pregenomic RNA on viral DNA synthesis. AB - The viral polymerase and several cis-acting sequences are essential for hepadnaviral DNA replication, but additional host factors are likely to be involved in this process. We previously identified two sequences, UBS and DBS (upstream and downstream binding sites), present in multiple copies in and adjacent to the pregenomic RNA (pgRNA) terminal redundancy, that were specifically recognized by a 65-kDa host factor, p65. The possible roles of these two sequences in hepatitis B virus (HBV) replication were investigated in the context of the intact viral genome. UBS is contained within the terminal redundancy of pgRNA, and the 5' copy of this sequence is essential for viral replication. Mutations within the central core of UBS ablate p65 binding and selectively block synthesis of plus-strand DNA, without affecting RNA packaging or minus-strand synthesis. The DBS sequence, which is located downstream of the pgRNA polyadenylation site, overlaps the core (C) protein coding region. All mutations introduced into this site severely affected viral replication. However, these effects were shown to result from dominant negative effects of mutant core polypeptides rather than from cis-acting effects on RNA recognition. Thus, the 5' UBS but not DBS sites play important cis-acting roles in HBV DNA replication; however, the involvement of p65 in these roles remains a matter for investigation. PMID- 9343202 TI - Mutational analysis of human immunodeficiency virus type 1 (HIV-1) accessory genes: requirement of a site in the nef gene for HIV-1 replication in activated CD4+ T cells in vitro and in vivo. AB - Human immunodeficiency virus type 1 (HIV-1) accessory genes including nef, vif, and vpr are important factors that determine the replication and pathogenesis of HIV-1. The state of activation is also important for the replication of HIV-1. We evaluated the properties of nef-, vif-, and vpr-minus macrophage-tropic HIV-1(JR) CSF in primary CD4+ Th1- or Th2-like cell cultures which had been activated through CD3 molecules in the presence of interleukin-2 (IL-2) and IL-12 (Th1-like culture) or IL-4 (Th2-like culture), respectively. In activated Th1- or Th2-like cultures, replication of nef-minus HIV-1(JR-CSF) was markedly lower than that of wild-type HIV-1. Subsequent analysis by site-directed mutagenesis showed that (i) the presence of an acidic amino acid-rich domain (amino acid residues 72 to 75) in the Nef protein was critical for the enhancement of viral DNA synthesis, resulting in increased virus growth rate, and (ii) prolines that form part of Src homology 3 binding domain were not essential for viral replication. We also confirmed the importance of sites by using an HIV-1-infected animal model, the hu PBL-SCID mouse system, representing HIV-1 replication and pathogenesis in activated CD4+ T cells in vivo. These results indicate that Nef accelerates viral replication in activated CD4+ T cells. PMID- 9343203 TI - Recombinant Listeria monocytogenes vaccination eliminates papillomavirus-induced tumors and prevents papilloma formation from viral DNA. AB - Listeria monocytogenes is a gram-positive, facultative intracellular bacterium that enters the cytoplasm of infected cells and spreads directly into neighboring cells without encountering the extracellular environment. Cytoplasmic L. monocytogenes efficiently presents secreted proteins to the major histocompatibility complex class I pathway which can stimulate protective T-cell mediated immune responses. We have used a cottontail rabbit papillomavirus (CRPV) rabbit model to test the ability of recombinant L. monocytogenes strains secreting the viral E1 protein (E1-rLm) to protect outbred rabbits against CRPV- and CRPV DNA-induced tumors. CRPV infection of outbred rabbits serves as a model for oncogenic papillomaviruses since CRPV-induced papillomas progress with high frequency to malignant carcinoma. Rabbits were vaccinated with wild-type L. monocytogenes or E1-rLm and then challenged with CRPV or viral DNA. In contrast to 0% papilloma regression in control animals, 77% of E1-rLm-vaccinated rabbits generated protective immunity that controlled and induced complete regression of tumors induced by CRPV. Latent viral DNA was not detected at 71% of the papilloma regression sites examined 4.5 months postregression. E1-rLm responder rabbits were completely resistant to papilloma formation from viral DNA. In contrast to controls, peripheral blood mononuclear cells from E1-rLm responder rabbits were able to proliferate in response to in vitro E1 stimulation. These results indicate that E1-rLm immunization generated a systemic anti-CRPV E1 cell-mediated immune response which protected outbred rabbits from tumors induced by CRPV or CRPV DNA challenge. PMID- 9343204 TI - Proteolytic activation of tick-borne encephalitis virus by furin. AB - Flaviviruses are assembled intracellularly in an immature form containing heterodimers of two envelope proteins, E and prM. Shortly before the virion exits the cell, prM is cleaved by a cellular enzyme, and this processing step can be blocked by treatment with agents that raise the pH of exocytic compartments. We carried out in vivo and in vitro studies with tick-borne encephalitis (TBE) virus to investigate the possible role of furin in this process as well as the functional consequences of prM cleavage. We found that prM in immature virions can be correctly cleaved in vitro by recombinant bovine furin but that efficient cleavage occurs only after exposure of the virion to mildly acidic pH. The data suggest that exposure to an acidic environment induces an irreversible structural change that renders the cleavage site accessible to the enzyme. Cleavage by furin in vitro resulted in biological activation, as shown by a 100-fold increase in specific infectivity, the acquisition of membrane fusion and hemagglutination activity, and the ability of the envelope proteins to undergo low-pH-induced structural rearrangements characteristic of mature virions. In vivo, prM cleavage was blocked by a furin inhibitor, and infection of the furin-deficient cell line LoVo yielded only immature virions, suggesting that furin is essential for cleavage activation of flaviviruses. PMID- 9343205 TI - Synchronous replication of poliovirus RNA: initiation of negative-strand RNA synthesis requires the guanidine-inhibited activity of protein 2C. AB - We report that protein 2C, the putative nucleoside triphosphatase/helicase protein of poliovirus, is required for the initiation of negative-strand RNA synthesis. Preinitiation RNA replication complexes formed upon the translation of poliovirion RNA in HeLa S10 extracts containing 2 mM guanidine HCI, a reversible inhibitor of viral protein 2C. Upon incubation in reactions lacking guanidine, preinitiation RNA replication complexes synchronously initiated and elongated negative-strand RNA molecules, followed by the synchronous initiation and elongation of positive-strand RNA molecules. The immediate and exclusive synthesis of negative-strand RNA upon the removal of guanidine demonstrates that guanidine specifically blocks the initiation of negative-strand RNA synthesis. Readdition of guanidine HCl to reactions synchronously elongating nascent negative-strand RNA molecules did not prevent their continued elongation and completion. In fact, readdition of guanidine HCl to reactions containing preinitiation complexes elongating nascent negative-strand RNA molecules had no effect on subsequent positive-strand RNA synthesis initiation or elongation. Thus, the guanidine-inhibited function of viral protein 2C was not required for the elongation of negative-strand RNA molecules, the initiation of positive strand RNA molecules, or the elongation of positive-strand RNA molecules. The guanidine-inhibited function of viral protein 2C is required only immediately before or during the initiation of negative-strand RNA synthesis. We suggest that guanidine may block an irreversible structural maturation of protein 2C and/or RNA replication complexes necessary for the initiation of RNA replication. PMID- 9343206 TI - Analysis of the murine leukemia virus R peptide: delineation of the molecular determinants which are important for its fusion inhibition activity. AB - In previous studies, the C-terminal R peptide of the murine leukemia virus (MuLV) Env protein was shown to be a potent inhibitor of viral fusion activity. In the present study, we investigated the molecular determinants in the MuLV Env protein cytoplasmic tail which are important for the fusion inhibition activity of the R peptide. We constructed a series of mutant MuLV env genes which express Env proteins with serial truncations, internal deletions, or amino acid substitutions in the cytoplasmic tail. To analyze their cell fusion activity, we employed a quantitative fusion assay. We found that truncations of up to 7 amino acids from the C terminus of the cytoplasmic tail had no detectable effect on the lack of fusion activity of the full-length Env protein; however, further truncations resulted in a progressive increase in cell fusion activity. Studies of mutant proteins with amino acid substitutions in the cytoplasmic tail showed that Leu 627 plays an important role in fusion inhibition by the R peptide, while most of the other amino acids in the R peptide were not essential for fusion inhibition. Studies of mutant proteins with internal deletions upstream of the cleavage site in the cytoplasmic tail showed that this region is also involved in fusion inhibition by the R peptide, although only to a limited extent. The results are consistent with a model in which the MuLV R peptide exhibits its fusion inhibition activity through interaction with a cellular factor(s). PMID- 9343207 TI - DNA immunization: ubiquitination of a viral protein enhances cytotoxic T lymphocyte induction and antiviral protection but abrogates antibody induction. AB - DNA immunization can induce cytotoxic T lymphocytes (CTL), antibodies, and protection against microbial challenge. The underlying mechanisms remain obscure and must be understood to permit rational manipulation and optimization of the technique. We set out to enhance the intracellular degradation of a viral antigen, with the intent of improving antigen entry into, and presentation by, the class I major histocompatibility complex pathway. We achieved this goal by cotranslational ubiquitination of a plasmid-encoded viral antigen, lymphocytic choriomeningitis virus (LCMV) nucleoprotein (NP). We show that native NP is very stable in cell culture, while the ubiquitinated product is so rapidly degraded that it is barely detectable. This rapid degradation leads to more efficient sensitization of target cells in an in vitro cytotoxicity assay, consistent with enhanced antigen presentation, and both degradation and target cell recognition are blocked by a proteasome inhibitor. We have used the plasmid for in vivo studies and find that, remarkably, ubiquitination leads to a complete abrogation of antibody responses, presumably because the encoded protein is so rapidly and completely degraded that insufficient antigen remains to interact appropriately with B cells. In contrast, in vivo CTL induction is improved by ubiquitination of NP. That CTL are induced at all by this rapidly degraded protein may shed light on the mechanism by which CTL are induced by DNA immunization; it has been suggested that CTL induction following intramuscular DNA injection results not from antigen presentation by cells taking up and expressing the DNA but rather from uptake of soluble protein by specialized antigen-presenting cells (APC). It appears to us unlikely that the ubiquitinated protein could function in this manner, since it is so rapidly degraded in vitro and fails to induce antibodies in vivo. Finally, the ubiquitinated protein confers markedly enhanced protection against LCMV challenge. Mice immunized with a plasmid encoding NP show approximately 100-fold reductions in virus titers compared to controls, while mice immunized with a plasmid encoding the ubiquitinated NP show reductions in virus load of at least 5 x 10(4)- to 5 x 10(5)-fold. This is by far the most effective DNA vaccine that we have yet designed. Ubiquitination therefore may improve DNA immunization, but caution is warranted, since immunity to many microbes depends on induction of good humoral immunity, and we show here that this may be prevented by ubiquitination of the encoded protein. PMID- 9343208 TI - Homologous sequences in the Campoletis sonorensis polydnavirus genome are implicated in replication and nesting of the W segment family. AB - Polydnaviruses (PDVs) are double-stranded DNA viruses with segmented genomes that replicate only in the oviducts of some species of parasitic wasps and are required for the successful parasitization of lepidopteran insects. PDV DNA segments are integrated in the genomes of their associated wasp hosts, and some are nested; i.e., smaller segments are produced from and largely colinear with larger segments. To determine the internal structure of nested viral segments, the first complete nucleotide sequence of a PDV genome segment and its integration locus was determined. By restriction mapping, Southern blot, and sequence analyses, we demonstrated that the Campoletis sonorensis PDV segment W is integrated into wasp genomic DNA. DNA sequence analysis revealed that proviral segment W terminates in two 1,185-bp direct long terminal repeats (LTRs) in the wasp chromosome, while only one LTR copy is present in the extrachromosomal (viral) W. The results suggest that terminal direct repeats are a general feature of PDV DNA segment integration but that the homology and size of the repeats can vary extensively. Segment W contains 12 imperfect direct repeats of six different types between 89 bp and 1.9 kbp with 65 to 90% homology. The orientation and structure of the repeats suggest that W itself may have arisen through sequence duplication and subsequent divergence. Mapping, hybridization, and sequence analyses of cloned R and M demonstrated that these segments are nested within segment W and that internal imperfect direct repeats of one type are implicated in the homologous intramolecular recombination events that generate segments R and M. Interestingly, segment nesting differentially increases the copy number of genes encoded by segment W, suggesting that the unusual genomic organization of PDVs may be directly linked to the unique functions of this virus in its obligate mutualistic association with parasitic wasps. PMID- 9343209 TI - The human T-cell leukemia virus type 1 Rex regulatory protein exhibits an impaired functionality in human lymphoblastoid Jurkat T cells. AB - The Rex protein of human T-cell leukemia virus type 1 (HTLV-1) intervenes in the posttranscriptional regulation of proviral gene expression. Its binding to the Rex response element (XRE) present in the 3' long terminal repeat ensures the coordinate cytoplasmic accumulation of spliced and unspliced forms of viral messengers. Consequently, synthesis of viral structural and enzymatic proteins is strictly dependent on the Rex posttranscriptional activity. Here we report that synthesis of HTLV-1 envelope glycoproteins by Jurkat T cells could be detected only when they were regulated in a Rex-independent manner. Indeed, Jurkat T cells transfected with a Rex-dependent env expression vector (encompassing both the env and pX open reading frames) do not produce significant levels of envelope glycoproteins despite the production of significant amounts of Rex protein. The analysis of levels and distribution patterns of the unspliced env and of the singly spliced tax/rex transcripts suggests that the failure in envelope glycoprotein synthesis may be ascribed to a deficiency of Rex in mediating the nucleocytoplasmic transport of unspliced env RNAs in these cells. Furthermore, despite the synthesis of regulatory proteins, HTLV-1 structural proteins were not detected in Jurkat T cells transfected with an HTLV-1 infectious provirus. Conversely, and as expected, structural proteins were produced by Jurkat cells transfected by a human immunodeficiency virus type 1 (HIV-1) infectious provirus. This phenotype appeared to be linked to a specific dysfunction of Rex, since the functionally equivalent Rev protein of HIV-1 was shown to be fully efficient in promoting the synthesis of HTLV-1 envelope glycoproteins in Jurkat cells. Therefore, it seems likely that the block to Rex function in these lymphoblastoid T cells is determined by inefficient Rex-XRE interactions. These observations suggest that the acquisition of this Rex-deficient phenotype by in vivo-infected HTLV-1 T cells may represent a critical event in the lymphoproliferation induced by this human retrovirus, leading to leukemia. PMID- 9343210 TI - Transcriptional activation of the vascular cell adhesion molecule-1 gene in T lymphocytes expressing human T-cell leukemia virus type 1 Tax protein. AB - Recruitment and extravasation of T cells through the blood-brain barrier are favored by adhesion molecule-mediated interactions of circulating T cells with endothelial cells. Since a common pathological finding in human T-cell leukemia virus type 1 (HTLV-1)-associated diseases is the infiltration of HTLV-1-infected T lymphocytes into various organs, we have looked for the profile of adhesion molecules expressed by HTLV-1-transformed T cells. Flow cytometry analysis indicated that these cells were expressing high levels of vascular cell adhesion molecule 1 (VCAM-1 [CD106]), a 110-kDa member of the immunoglobulin gene superfamily, first identified on endothelial cells stimulated with inflammatory cytokines. This adhesion molecule was also expressed by T cells obtained from one patient with HTLV-1-associated myelopathy/tropical spastic paraparesis but not by activated T cells isolated from one normal blood donor. The role of the viral trans-activator Tax protein in the induction of VCAM-1 was first indicated by the detection of this adhesion molecule on Jurkat T-cell clones stably expressing the tax gene. The effect of Tax on VCAM-1 gene transcription was next confirmed in JPX-9 cells, a subclone of Jurkat cells, carrying the tax sequences under the control of an inducible promoter. Furthermore, deletion and mutation analyses of the VCAM-1 promoter performed with chloramphenicol acetyltransferase constructs revealed that Tax was trans activating the VCAM-1 promoter via two NF-kappaB sites present at bp -72 and -57 in the VCAM-1 gene promoter, with both of them being required for the Tax-induced expression of this adhesion molecule. Finally, gel mobility shift assays demonstrated the nuclear translocation of proteins specifically bound to these two NF-kappaB motifs, confirming that VCAM-1 was induced on Tax-expressing cells in a kappaB-dependent manner. Collectively, these results therefore suggest that the exclusive Tax-induced expression of VCAM-1 on T cells may represent a pivotal event in the progression of HTLV-1-associated diseases. PMID- 9343212 TI - A naturally arising mutation of a potential silencer of exon splicing in human immunodeficiency virus type 1 induces dominant aberrant splicing and arrests virus production. AB - We have isolated a naturally arising human immunodeficiency type 1 (HIV-1) mutant containing a point mutation within the env gene. The point mutation resulted in complete loss of balanced splicing, with dominant production of aberrant mRNAs. The aberrant RNAs arose via activation of normally cryptic splice sites flanking the mutation within the env terminal exon to create exon 6D, which was subsequently incorporated in aberrant env, tat, rev, and nef mRNAs. Aberrant multiply spliced messages contributed to reduced virus replication as a result of a reduction in wild-type Rev protein. The point mutation within exon 6D activated exon 6D inclusion when the exon and its flanking splice sites were transferred to a heterologous minigene. Introduction of the point mutation into an otherwise wild-type HIV-1 proviral clone resulted in virus that was severely inhibited for replication in T cells and displayed elevated usage of exon 6D. Exon 6D contains a bipartite element similar to that seen in tat exon 3 of HIV-1, consisting of a potential exon splicing silencer (ESS) juxtaposed to a purine-rich sequence similar to known exon splicing enhancers. In the absence of a flanking 5' splice site, the point mutation within the exon 6D ESS-like element strongly activated env splicing, suggesting that the putative ESS plays a natural role in limiting the level of env splicing. We propose, therefore, that exon silencers may be a common element in the HIV-1 genome used to create balanced splicing of multiple products from a single precursor RNA. PMID- 9343211 TI - An adenovirus-simian immunodeficiency virus env vaccine elicits humoral, cellular, and mucosal immune responses in rhesus macaques and decreases viral burden following vaginal challenge. AB - Six female rhesus macaques were immunized orally and intranasally at 0 weeks and intratracheally at 12 weeks with an adenovirus type 5 host range mutant (Ad5hr) simian immunodeficiency virus SIVsm env recombinant and at 24 and 36 weeks with native SIVmac251 gp120 in Syntex adjuvant. Four macaques received the Ad5hr vector and adjuvant alone; two additional controls were naive. In vivo replication of the Ad5hr wild-type and recombinant vectors occurred with detection of Ad5 DNA in stool samples and/or nasal secretions in all macaques and increases in Ad5 neutralizing antibody in 9 of 10 macaques following Ad administrations. SIV-specific neutralizing antibodies appeared after the second recombinant immunization and rose to titers > 10,000 following the second subunit boost. Immunoglobulin G (IgG) and IgA antibodies able to bind gp120 developed in nasal and rectal secretions, and SIV-specific IgGs were also observed in vaginal secretions and saliva. T-cell proliferative responses to SIV gp140 and T-helper epitopes were sporadically detected in all immunized macaques. Following vaginal challenge with SIVmac251, transient or persistent infection resulted in both immunized and control monkeys. The mean viral burden in persistently infected immunized macaques was significantly decreased in the primary infection period compared to that of control macaques. These results establish in vivo use of the Ad5hr vector, which overcomes the host range restriction of human Ads for rhesus macaques, thereby providing a new model for evaluation of Ad-based vaccines. In addition, they show that a vaccine regimen using the Ad5hr-SIV env recombinant and gp120 subunit induces strong humoral, cellular, and mucosal immunity in rhesus macaques. The reduced viral burden achieved solely with an env-based vaccine supports further development of Ad-based vaccines comprising additional viral components for immune therapy and AIDS vaccine development. PMID- 9343214 TI - Receptor-targeted recombinant adenovirus conglomerates: a novel molecular conjugate vector with improved expression characteristics. AB - To develop improved strategies for gene transfer to hematopoietic cells, we have explored targeted gene transfer using molecular conjugate vectors (MCVs). MCVs are constructed by condensing plasmid DNA containing the gene of interest with polylysine (PL), PL linked to a replication-incompetent adenovirus (endosomolytic agent), and PL linked to streptavidin for targeting with biotinylated ligands. In this report, we compare gene transfer to K562 cells by using the previously described transferrin-targeted MCV (Trans-MCV) to a novel transferrin-targeted MCV. In the novel MCV, the transferred gene (luciferase) is in the genome of recombinant replication-incompetent adenovirus (recMCV), which also acts as the endosomolytic agent. The level of luciferase gene expression was fivefold higher in K562 cells transfected with Trans-recMCV than in cells transfected with Trans MCV. Furthermore, targeted transfection with recMCV resulted in prolonged luciferase expression that declined 14 to 20 days after transfection, in comparison with Trans-MCV, where luciferase expression declined by 4 to 8 days. Moreover, targeted transfection of K562 cells with the Trans-recMCV resulted in persistent luciferase gene expression for 6 months. Analysis of luciferase gene expression in K562 single-cell clones that were subcloned 5 weeks after transfection with Trans-recMCV showed that 35 to 50% of the single-cell clones had intermediate to high levels of luciferase gene expression that was stable for 6 months, with the remaining clones showing low or no luciferase gene expression. Stable gene expression was associated with integration of adenovirus sequences into genomic DNA. PMID- 9343213 TI - Epstein-Barr virus EBNA3C represses Cp, the major promoter for EBNA expression, but has no effect on the promoter of the cell gene CD21. AB - EBNA3C is a potent repressor of transcription when bound to DNA as a fusion with the DNA binding domain (DBD) of GALA. A survey of promoters has revealed that the wild-type, unfused EBNA3C can specifically repress expression from reporter plasmids containing the Epstein-Barr virus Cp latency-associated promoter. Repression of Cp activity required amino acids 207 to 368, which encompasses a region resembling a basic DBD adjacent to a leucine zipper DNA binding motif and a site which binds to the cellular factor CBF1/RBP-Jkappa. However, amino acids 207 to 368 are dispensable when the protein is bound to DNA as a fusion with the GAL4 DBD, thus implicating this region in DNA binding. Mutation of the CBF1/RBP Jkappa binding site in EBNA3C abrogated repression, strongly suggesting that CBF1/RBP-Jkappa is necessary for targeting the viral protein to Cp. Consistent with this result, mutation of the EBNA2 response element (a CBF1/RBP-Jkappa binding site) in Cp also prevented significant repression. In addition, amino acids 346 to 543, which were previously defined as important for the repressor activity of the GAL4-EBNA3C fusion proteins, also appear to be necessary for the repression of Cp. Since repression by these fusions was not observed in all cell types, it seems likely that EBNA3C either depends on a corepressor which may interact with amino acids 346 to 543 or is modified in a cell-specific manner in order to repress. These data are consistent with EBNA3C contributing to the regulation of EBNA expression in latently infected B cells through CBF1/RBP Jkappa and another factor, but this need not directly involve EBNA2. Finally, although it has been reported that EBNA3C can upregulate CD21 in some B cells, we were unable to demonstrate any effect of EBNA3C on reporter plasmids which contain the CD21 promoter. PMID- 9343215 TI - Alterations in catalytic activity and virus maturation produced by mutation of the conserved histidine residues of herpes simplex virus type 1 protease. AB - Mutant herpes simplex virus type 1 (HSV-1) viruses were constructed to characterize the roles of the conserved histidine residues (H61 and H148) of HSV 1 protease in the regulation of catalytic activity and virus maturation. Viruses containing mutations at H61 (H61V-V711, H61Y-V715, and H61A-V730) were unable to grow on Vero cells. These mutant viruses could process neither Pra to N0 nor ICP 35cd to ICP-35ef. Transmission electron microscopy studies of H61A-V730-infected Vero cells indicated that capsid maturation is arrested at a state characterized by the predominance of large symmetrical arrays of B capsids within the nucleus. Two mutations at H148 (in viruses H148A-V712 and H148E-V728) gave rise to mutant viruses that grew with a small-plaque phenotype; one of the viruses, H148E-V728, was particularly attenuated when grown at a low multiplicity of infection. The rate of processing of Pra to N0 in infected Vero cells increased in the order H148A-V712 < H148E-V728 < parental strain HSV-1-V731. The observation that H148A V712 processes Pra to N0 and ICP-35cd to ICP-35ef, whereas H61A does not, establishes H61 as the catalytically essential conserved His assuming that HSV-1 protease, like other serine proteases, utilizes an active-site histidine residue in catalysis. Two of the mutations at H148 (viruses H148K-V729 and H148Y-V716) produced nonviable viruses. H148K-V729 processed neither Pra to N0 nor ICP-35cd to ICP-35ef, whereas H148Y-V716 processed Pra to N0 but did not process ICP-35cd to ICP-35ef. The range of phenotypes observed with the H148 mutant viruses suggests that residue 148 of the HSV-1 protease is a determinant of virus growth rate and viability because of its effects on the activity of the protease and/or the role of the protease domain in capsid assembly and DNA packaging. PMID- 9343216 TI - Immunoglobulin V(H) usage during primary infection of rhesus monkeys with chimeric simian-human immunodeficiency viruses. AB - It has been suggested that naive immunoglobulins encoded by the V(H)3 gene family interact aberrantly with human immunodeficiency virus type 1 (HIV-1) gp120 via a superantigenic epitope, causing initial expansion and eventual depletion of V(H)3 expressing B cells. However, this possibility has not been prospectively assessed during an AIDS virus infection. We determined V(H) family usage in rhesus monkeys during primary infection with chimeric viruses expressing HIV-1 envelopes on a simian immunodeficiency virus (SIVmac) backbone (SHIVs). Four SHIVs with different envelopes and pathogenicities were studied. V(H) family usage was prospectively assessed in peripheral blood mononuclear cells and lymph node cells of these monkeys by a semiquantitative PCR technique. In the first months following SHIV infection, a period of intense viral antigenemia, representation of various V(H) families increased or decreased for individual monkeys, but no single V(H) family was consistently altered. In particular, the average representation of V(H)3-bearing B lymphocytes did not change. This observation suggests that the envelope glycoprotein of HIV-1 does not selectively expand or deplete the V(H)3 repertoire of primate B cells during acute AIDS virus infection, contrary to predictions of the gp120 superantigen hypothesis. PMID- 9343217 TI - The immune system preferentially clears Theiler's virus from the gray matter of the central nervous system. AB - Infection of susceptible strains of mice with Daniel's (DA) strains of Theiler's murine encephalomyelitis virus (DAV) results in virus persistence in the central nervous system (CNS) white matter and chronic demyelination similar to that observed in multiple sclerosis. We investigated whether persistence is due to the immune system more efficiently clearing DAV from gray than from white matter of the CNS. Severe combined immunodeficient (SCID) and immunocompetent C.B-17 mice were infected with DAV to determine the kinetics, temporal distribution, and tropism of the virus in CNS. In early disease (6 h to 7 days postinfection), DAV replicated with similar kinetics in the brains and spinal cords of SCID and immunocompetent mice and in gray and white matter. DAV RNA was localized within 48 h in CNS cells of all phenotypes, including neurons, oligodendrocytes, astrocytes, and macrophages/microglia. In late disease (13 to 17 days postinfection), SCID mice became moribund and permitted higher DAV replication in both gray and white matter. In contrast, immunocompetent mice cleared virus from the gray matter but showed replication in the white matter of their brains and spinal cords. Reconstitution of SCID mice with nonimmune splenocytes or anti-DAV antibodies after establishment of infection demonstrated that both cellular and humoral immune responses decreased virus from the gray matter; however, the cellular responses were more effective. SCID mice reconstituted with splenocytes depleted of CD4+ or CD8+ T lymphocytes cleared virus from the gray matter but allowed replication in the white matter. These studies demonstrate that both neurons and glia are infected early following DAV infection but that virus persistence in the white matter is due to preferential clearance of virus from the gray matter by the immune system. PMID- 9343218 TI - Mutational analysis of the herpes simplex virus type 1 ICP0 C3HC4 zinc ring finger reveals a requirement for ICP0 in the expression of the essential alpha27 gene. AB - The herpes simplex virus type 1 (HSV-1) immediate-early (IE) protein ICP0 has been implicated in the regulation of viral gene expression and the reactivation of latent HSV-1. Evidence demonstrates that ICP0 is an activator of viral gene expression yet does not distinguish between a direct or indirect role in this process. To further our understanding of the function of ICP0 in the context of the virus life cycle, site-directed mutagenesis of the consensus C3HC4 zinc finger domain was performed, and the effects of these mutations on the growth and replication of HSV-1 were assessed. We demonstrate that alteration of any of the consensus C3HC4 cysteine or histidine residues within this domain abolishes ICP0 mediated transactivation, alters the intranuclear localization of ICP0, and significantly increases its stability. These mutations result in severe defects in the growth and DNA replication of recombinant herpesviruses and in their ability to initiate lytic infections at low multiplicities of infection. These viruses, at low multiplicities of infection, synthesize wild-type levels of the IE proteins ICP0 and ICP4 at early times postinfection yet exhibit significant decreases in the synthesis of the essential IE protein ICP27. These findings reveal a role for ICP0 in the expression of ICP27 and suggest that the multiplicity-dependent growth of alpha0 mutant viruses results partially from reduced levels of ICP27. PMID- 9343219 TI - Cytotoxic T-lymphocyte cross-reactivity among different human immunodeficiency virus type 1 clades: implications for vaccine development. AB - Despite recent advances in antiviral therapy for human immunodeficiency virus (HIV) infection, successful global intervention will require an effective vaccine. Expanding evidence suggests that cytotoxic T-lymphocyte (CTL) responses will be an important component of such a vaccine. The varying geographic distribution of HIV type 1 (HIV-1) clades, with the relative absence of clade B HIV-1 outside the developed world, is considered a major obstacle to the development of a single efficacious vaccine. An understanding of cross-reactive CTL responses between different HIV-1 clades is crucial in the design of a vaccine which will be broadly immunogenic. In this study, we examined the ability of HIV-1 Gag-, reverse transcriptase-, and Env-specific CTL clones isolated from individuals infected in the United States to recognize non-B clade viral sequences and found that all were cross-reactive with the majority of non-B clade viral sequences tested. We next studied HIV-1-specific CTL responses in African individuals infected with clade A, C, or G virus and evaluated cross-recognition of clade B virus. Of 14 persons evaluated, all demonstrated cross-reactivity with the U.S. clade B viral constructs. We conclude that significant CTL cross reactivity exists between clade B and non-B epitopes, suggesting that CTL cross recognition among HIV-1 clades is more widespread than anticipated and that a vaccine based on a single clade may be broadly applicable. PMID- 9343220 TI - VPg of tobacco etch potyvirus is a host genotype-specific determinant for long distance movement. AB - The V20 cultivar of Nicotiana tabacum was shown previously to exhibit a strain specific restriction of long-distance movement of tobacco etch potyvirus (TEV). In V20, both TEV-HAT and TEV-Oxnard strains are capable of genome amplification and cell-to-cell movement, but only TEV-Oxnard is capable of systemic infection by vasculature-dependent long-distance movement. To investigate the basis for host-specific movement of TEV, chimeric virus genomes were assembled from TEV-HAT and TEV-Oxnard. Viruses containing the TEV-Oxnard coding regions for HC-Pro and/or capsid protein (CP), two proteins that are known to be essential for TEV long-distance movement, failed to infect V20 systemically. In contrast, chimeric viruses encoding the TEV-Oxnard VPg domain of NIa were able to infect V20 systemically. The critical region controlling the infection phenotype in V20 was mapped to a 67-nucleotide segment containing 10-nucleotide differences, but only five amino acid differences, between TEV-HAT and TEV-Oxnard. In V20 coinfection experiments, a restricted strain had no effect on systemic infection by a long distance movement-competent chimeric strain, suggesting that the restricted strain was not inducing a generalized systemic resistance response. These data suggest that the VPg domain, which is covalently attached to the 5' end of genomic RNA, interacts either directly or indirectly with host components to facilitate long-distance movement. PMID- 9343221 TI - Immunopathologic changes in the thymus during the acute stage of experimentally induced feline immunodeficiency virus infection in juvenile cats. AB - The feline thymus is a target organ and site of viral replication during the acute stage of feline immunodeficiency virus (FIV) infection. This was demonstrated by histologic, immunohistologic, flow cytometric, and virologic tests. Thymic lesions developed after 28 days postinoculation (p.i.) and included thymitis, premature cortical involution, and medullary B-cell hyperplasia with germinal center formation and epithelial distortion. Alterations in thymocyte subsets also developed. Fewer CD4+ CD8- cells were detected at 28 days p.i., while an increase in CD4- CD8+ cells resulted in an inversion of the thymic CD4/CD8 ratio of single-positive cells, similar to events in peripheral blood. Provirus was present in all thymocyte subpopulations including cortical CD1(hi), CD1(lo), and B cells. The CD1(hi) thymocyte proviral burden increased markedly after 56 days p.i., coincident with the presence of infiltrating inflammatory cells. Increased levels of provirus in the CD1(lo) thymocyte subpopulation were detected prior to 56 days p.i. This was likely due to inclusion of infected infiltrating inflammatory cells which could not be differentiated from mature, medullary thymocytes. Proviral levels in B cells also increased from 70 days p.i. Morphologic alterations, productive viral infection, and altered thymocyte subpopulations suggest that thymic function is compromised, thus contributing to the inability of FIV-infected cats to replenish the peripheral T-cell pool. PMID- 9343222 TI - Polymorphisms in the CCR5 genes of African green monkeys and mice implicate specific amino acids in infections by simian and human immunodeficiency viruses. AB - CCR5, a receptor for the CC chemokines RANTES, Mip1alpha, and Mip1beta, has been identified as a coreceptor for infections by macrophage-tropic isolates of human immunodeficiency virus type 1 (HIV-1). To study its structure and function, we isolated cDNA clones of human, African green monkey (AGM), and NIH/Swiss mouse CCR5s, and we quantitatively analyzed infections by macrophage-tropic HIV-1 and SIVmac251 after transfecting human HeLa-CD4 cells with the CCR5 expression vectors. The AGM and NIH/Swiss mouse CCR5 proteins are 97.7 to 98.3% and 79.8% identical to the human protein, respectively. In addition, we analyzed site directed mutants and chimeras of these CCR5s. Cell surface expression of CCR5 proteins was monitored by using a specific rabbit antiserum and by binding the chemokine [125I]Mip1beta. Our major results were as follows. (i) Two distinct AGM CCR5 sequences were reproducibly found in DNA from CV-1 cells. The AGM clone 1 CCR5 protein differs from that of clone 2 by two substitutions, Y14N in the amino terminal extracellular region and L352F at the carboxyl terminus. Interestingly, AGM clone 1 CCR5 was inactive as a coreceptor for all tested macrophage-tropic isolates of HIV-1, whereas AGM clone 2 CCR5 was active. As shown by chimera studies and site-directed mutagenesis, the Y14N substitution in AGM clone 1 CCR5 was solely responsible for blocking HIV-1 infections. In contrast, both AGM CCR5 clones were active coreceptors for SIVmac251. Studies of DNA samples from other AGMs indicated frequent additional CCR5 polymorphisms, and we cloned an AGM clone 2 variant with a Q93R substitution in the extracellular loop 1 from one heterozygote. This variant CCR5 was active as a coreceptor for SIVmac251 but was only weakly active for macrophage-tropic isolates of HIV-1. In addition, SIVmac251 appeared to be dependent on the extracellular amino terminus and loop 2 regions of human CCR5 for maximal infection. Our results suggest major differences in the interactions of SIVmac251 and macrophage-tropic HIV-1 isolates with 19, N13, and Y14 in the amino terminus; with Q93 in extracellular loop 1; and with extracellular loop 2 of human CCR5. (ii) The NIH/Swiss mouse CCR5 protein differs at multiple positions from sequences recently reported for other inbred strains of mice. This CCR5 was inactive as a coreceptor for HIV-1 and SIVmac251. Studies of chimeras that contained different portions of NIH/Swiss mouse CCR5 substituted into human CCR5, as well as the reciprocal chimeras, indicated that the amino-terminal region and extracellular loops 1 and 2 of human CCR5 contribute to its coreceptor activity for macrophage-tropic isolates of HIV 1. Specific differences with previous CCR5 chimera results occurred because the NIH/Swiss mouse CCR5 contains a unique substitution corresponding to P183L in extracellular loop 2 that is nonpermissive for coreceptor activity. We conclude that diverse CCR5 sequences occur in AGMs and mice, that SIVmac251 and macrophage tropic HIV-1 isolates interact differently with specific CCR5 amino acids, and that multiple regions of human CCR5 contribute to its coreceptor functions. In addition, we have identified naturally occurring amino acid polymorphisms in three extracellular regions of CCR5 (Y14N, Q93R, and P183L) that do not interfere with cell surface expression or Mip1beta binding but prevent infections by macrophage-tropic isolates of HIV-1. In contrast to previous evidence, these results suggest that CCR5 contains critical sites that are essential for HIV-1 infections. PMID- 9343224 TI - Characterization of human serotype 1 astrovirus-neutralizing epitopes. AB - Astroviruses are important agents of pediatric gastroenteritis. To better understand astrovirus antigenic structure and the basis of protective immunity, monoclonal antibodies (MAbs) were produced against serotype 1 human astrovirus. Four MAbs were generated. One MAb (8G4) was nonneutralizing but reacted to all seven serotypes of astrovirus by enzyme-linked immunosorbentassay (ELISA) and immunoperoxidase staining of infected cells. Three MAbs were found to have potent neutralizing activity against astrovirus. The first (5B7) was serotype 1 specific, another (7C2) neutralized all seven human astrovirus serotypes, while the third (3B2) neutralized serotypes 1 and 7. Immunoprecipitation of radiolabeled astrovirus proteins from supernatants of astrovirus-infected cells showed that all three neutralizing antibodies reacted with VP29. MAb 5B7 also reacted strongly with VP26. A competition ELISA showed that all three neutralizing antibodies competed with each other for binding to purified astrovirus virions, suggesting that their epitopes were topographically in close proximity. None of the neutralizing MAbs competed with nonneutralizing MAb 8G4. The neutralizing MAbs were used to select antigenic variant astroviruses, which were then studied in neutralization assays. These assays also suggested a close relationship between the respective epitopes. All three neutralizing MAbs were able to prevent attachment of radiolabeled astrovirus particles to human Caco 2 intestinal cell monolayers. Taken together, these data suggest that the astrovirus capsid protein VP29 may be important in viral neutralization, heterotypic immunity, and virus attachment to target cells. PMID- 9343223 TI - Divergent transcriptional regulation among expanding human immunodeficiency virus type 1 subtypes. AB - The current AIDS pandemic represents the uneven spread of multiple genetically related subtypes (A to J) of human immunodeficiency virus type 1 (HIV-1). Notably, HIV-1 E in southeast Asia and HIV-1 C in sub-Saharan Africa are expanding faster and are likely of greater global significance than the HIV-1 B subtype prevalent in the United States and Europe. While many studies have focused on genetic variation among structural genes, we chose to conduct a comparative analysis of the long terminal repeats of HIV-1 E and HIV-1 C isolates and report subtype-specific differences in enhancer copy numbers and sequences, as well as divergent activation in response to the cellular transcriptional activators Rel-p65 and NFATc and viral Tat. This study is the first to identify functional distinctions in promoter architecture between HIV-1 subtypes and raises the possibility that regulatory divergence among the subtypes of HIV-1 has occurred. Divergent transcriptional regulation may explain some of the epidemiologically observed differences in transmission and pathogenesis and underscores the need for further comparative analysis of HIV-1 regulation. PMID- 9343225 TI - Overexpression of interleukin-4 delays virus clearance in mice infected with respiratory syncytial virus. AB - Although interleukin-4 (IL-4) expression has been implicated in vaccine-enhanced respiratory syncytial virus (RSV) disease, its role in mediating the immune response to primary RSV infection remains unclear. To assess the effect of IL-4 production on typical RSV infection, transgenic mice which either overexpress or fail to express IL-4 were challenged intranasally with RSV and their responses were compared to those of the parent strains. IL-4-deficient mice eliminated virus from the lung as quickly as did C57BL/6 controls. In contrast, mice which constitutively overexpress IL-4 showed delayed virus clearance compared with mice of the FVB/N control strain, although peak viral titers did not differ. IL-4 overexpression increased the magnitude of the subsequent antibody response. Lung lymphocytes harvested from IL-4-overexpressing mice post-RSV challenge showed diminished RSV-specific cytolytic activity compared with controls. Both IL-4 deficient and IL-4-overexpressing strains resisted rechallenge. These data imply that constitutive IL-4 expression delays or suppresses the development of a virus specific cytotoxic lymphocyte population important in clearing primary RSV infection. PMID- 9343226 TI - Structural and functional determinants in adenovirus type 2 penton base recombinant protein. AB - Discrete domains involved in structural and functional properties of adenovirus type 2 (Ad2) penton base were investigated with site-directed mutagenesis of the recombinant protein expressed in baculovirus-infected cells. Seventeen substitution mutants were generated and phenotyped for various functions in insect and human cells as follows. (i) Pentamerization of the penton base protein was found to be dependent on three amino acid side chains, the indole ring of Trp119, the hydroxylic group of Tyr553, and the basic group of Lys556. (ii) Arg254, Cys432, and Trp439, the stretch of basic residues at positions 547 to 556, and Arg340 of the RGD motif played a critical role in stable fiber-penton base interactions in vivo. (iii) Nuclear localization of penton base in Sf9 cells was negatively affected in mutants W119H or W165H, and, to a lesser extent, by substitutions in the consensus polybasic signal at positions 547 to 549. (iv) Penton base mutants were also assayed for HeLa cell binding, cell detachment, plasmid DNA internalization, and Ad-mediated gene delivery. The results obtained suggested that the previously identified integrin-binding motifs RGD340 and LDV287 were functionally and/or topologically related to other discrete regions which include Trp119, Trp165, Cys246, Cys432, and Trp439, all of which were involved in penton base-cell surface recognition, endocytosis, and postendocytotic steps of the virus life cycle. PMID- 9343227 TI - Differential recruitment of B and T cells in coxsackievirus B4-induced pancreatitis is influenced by a capsid protein. AB - Two genetically similar variants of coxsackievirus B4, CB4-P and CB4-V, cause distinct disease syndromes in mice. A multidisciplinary approach was used to examine the events occurring in situ. The CB4-P variant induced acute pancreatitis, followed by repair of the exocrine tissues, while the CB4-V variant induced chronic pancreatitis, characterized by extensive destruction of the exocrine tissues. Since CB4-V replicated more efficiently than CB4-P in vivo, the more extensive tissue injury associated with CB4-V infection could be explained as the result of a higher level of viral replication. However, the fact that CB4 V replicated more efficiently in a mouse strain that survives infection than in a strain that succumbs to infection suggests that immune-mediated mechanisms as well as viral cytolysis may contribute to pancreatic tissue injury. To address the role of the immune system in virus-induced pancreatitis, the cell types within the inflammatory infiltrate were analyzed by flow cytometry. B cells (34 to 75%) were the most abundant, followed by T cells (10 to 30%), natural killer cells (4 to 8%), and macrophages (0 to 6%). Recruitment (and perhaps proliferation) of B and T cells to the pancreatic tissues was influenced by viral strain. Differential recruitment of T and B cells may reflect altered antigenic sites between CB4-P and CB4-V. The viral sequence that affected T- and B-cell recruitment was identified as a threonine residue at position 129 of the VP1 capsid protein. PMID- 9343228 TI - Determination of the secondary structure of and cellular protein binding to the 3'-untranslated region of the hepatitis C virus RNA genome. AB - Hepatitis C virus (HCV) contains a positive-stranded RNA genome of approximately 9.5 kb. Despite the overall sequence diversity among individual HCV isolates, the 3'-end 98 nucleotides (nt) of the HCV RNA, which constitute part of the 3' untranslated region (3'-UTR), are highly conserved. This conserved region may contain the cis-acting signals for RNA replication involving possibly both viral and cellular proteins. We carried out RNase digestion studies, which revealed that this 98-nt region contains three stem-loops but may also assume alternative structures. We further performed UV cross-linking experiments to detect cellular proteins that bound to this region. A 58-kDa cellular protein (p58) was detected. Its binding site was mapped to the stem-loops 2 and 3, which are the most conserved region of the 3'-UTR. Site-directed mutagenesis studies revealed that both stem structures and specific nucleotide sequence within the two loops are important for p58 binding. Mutations that disrupted stem structures abolished protein binding, while the compensatory mutations restored its binding. This region also contains partial sequence similarity to the reported consensus binding sequence for polypyrimidine tract-binding protein (PTB) (a 57-kDa protein). The UV-cross-linked protein could be immunoprecipitated with the anti PTB antibody, and the recombinant PTB bound to the HCV 3'-UTR with the same binding specificity as p58, establishing that this protein is PTB. However, the reported PTB-binding sequence was not sufficient, but rather the entire stem loops 2 and 3 were required, for PTB binding; thus, its binding specificity is significantly different from the reported PTB-binding sequence requirement. This protein was detected in both the nuclei and cytoplasm of most mammalian cell lines tested and human primary hepatocytes. PTB may participate in the regulation of HCV RNA synthesis or translation. PMID- 9343229 TI - Rotavirus virus-like particles administered mucosally induce protective immunity. AB - We have evaluated the immunogenicity and protective efficacy of rotavirus subunit vaccines administered by mucosal routes. Virus-like particles (VLPs) produced by self-assembly of individual rotavirus structural proteins coexpressed by baculovirus recombinants in insect cells were the subunit vaccine tested. We first compared the immunogenicities and protective efficacies of VLPs containing VP2 and VP6 (2/6-VLPs) and G3 2/6/7-VLPs mixed with cholera toxin and administered by oral and intranasal routes in the adult mouse model of rotavirus infection. VLPs administered orally induced serum antibody and intestinal immunoglobulin A (IgA) and IgG. The highest oral dose (100 microg) of VLPs induced protection from rotavirus challenge (> or = 50% reduction in virus shedding) in 50% of the mice. VLPs administered intranasally induced higher serum and intestinal antibody responses than VLPs administered orally. All mice receiving VLPs intranasally were protected from challenge; no virus was shed after challenge. Since there was no difference in immunogenicity or protective efficacy between 2/6- and 2/6/7-VLPs, protection was achieved without inclusion of the neutralization antigens VP7 and VP4. We also tested the immunogenicities and protective efficacies of 2/6-VLPs administered intranasally without the addition of cholera toxin. 2/6-VLPs administered intranasally without cholera toxin induced lower serum and intestinal antibody titers than 2/6-VLPs administered with cholera toxin. The highest dose (100 microg) of 2/6-VLPs administered intranasally without cholera toxin resulted in a mean reduction in shedding of 38%. When cholera toxin was added, higher levels of protection were achieved with 10-fold less immunogen. VLPs administered mucosally offer a promising, safe, nonreplicating vaccine for rotavirus. PMID- 9343230 TI - cis-Acting signals involved in termination of vesicular stomatitis virus mRNA synthesis include the conserved AUAC and the U7 signal for polyadenylation. AB - We investigated the cis-acting sequences involved in termination of vesicular stomatis virus mRNA synthesis by using bicistronic genomic analogs. All of the cis-acting signals necessary for termination reside within the first 13 nucleotides of the 23-nucleotide conserved gene junction. This 13-nucleotide termination sequence at the end of the upstream gene comprises the tetranucleotide AUAC, the tract containing seven uridines (U7 tract), and the intergenic dinucleotide (GA), but it does not include the downstream gene start sequence. Data presented here show that upstream mRNA termination is independent of downstream mRNA initiation. Alteration of any nucleotide in the 13-nucleotide sequence decreased the termination activity of the gene junction and resulted in increased synthesis of a bicistronic readthrough RNA. This finding indicated that the wild-type gene junction has evolved to achieve the maximum termination efficiency. The most critical position of the AUAC sequence was the C, which could not be altered without complete loss of mRNA termination. Reducing the length of the wild-type U7 tract to zero, five, or six U residues also totally abolished mRNA termination, resulting in exclusive synthesis of the bicistronic readthrough mRNA. Shortening the wild-type U7 tract to either five or six U residues abolished VSV polymerase slippage during readthrough RNA synthesis. Since neither the U5 nor U6 template was able to direct mRNA termination, these data imply that polymerase slippage is a prerequisite for termination. Evidence is also presented to show that in addition to causing polymerase slippage, the U7 tract itself or its poly(A) product constitutes an essential signal for mRNA termination. PMID- 9343231 TI - Late gene expression from the Epstein-Barr virus BcLF1 and BFRF3 promoters does not require DNA replication in cis. AB - Late gene expression follows and is dependent upon lytic replication of the viral genome. Although experimental evidence is lacking, lytic viral DNA replication is believed to remove modifications or binding factors from the genome which serve to repress late gene expression during latency or the early lytic cycle. We have developed a reporter assay to begin characterizing the mechanisms that regulate late gene expression in Epstein-Barr virus (EBV). In this model system, the activities of late promoter-reporter fusions are measured following transient transfection into tissue culture cells expressing EBV during different stages of the lytic cycle. This system faithfully recapitulates late expression patterns from the endogenous virus, implicating specific cis-active sequences in the control of late gene expression. In addition, these promoters respond only indirectly to the viral immediate-early transactivator, ZEBRA. This indirect response is mediated by other viral or virally induced activities downstream of ZEBRA in the lytic cascade. In this system, late gene expression is sensitive to inhibitors of the viral DNA polymerase such as phosphonoacetic acid, although the reporters lack a eukaryotic origin of replication and are not replicated under the assay conditions. Thus, replication of the transcriptional template is not a prerequisite for expression with late kinetics, a finding inconsistent with the current models which posit a cis-active relationship between lytic EBV DNA replication and late gene expression. Rather, analysis of this system has revealed a trans relationship between late gene expression and viral DNA replication and highlights the indirect and complex link between these two events. PMID- 9343232 TI - Evidence that resolution of rotavirus infection in mice is due to both CD4 and CD8 cell-dependent activities. AB - The effector functions responsible for resolution of shedding in mice orally inoculated with the murine rotavirus strain EDIM were identified in B-cell deficient and normal BALB/c mice after monoclonal antibody (MAb) depletion of CD4 and CD8 cells. When depleted of CD8 cells, B-cell-deficient muMt mice resolved their infections more slowly than nondepleted animals, but CD4 cell depletion caused chronic, high-level shedding. This finding indicated that CD4 cell dependent immunological effectors other than, or in addition to, CD8 cells played roles in rotavirus resolution in muMt mice in the absence of antibody. The roles of CD4 and CD8 cells in resolution of rotavirus shedding were further characterized in immunologically normal BALB/c mice. Depletion of CD4 cells before EDIM inoculation resulted in rapid resolution of most shedding, but chronic, low-level shedding continued for weeks. When the CD4 cell-depleted BALB/c mice were subsequently depleted of CD8 cells, shedding levels increased significantly (P < 0.001), indicating that CD8 cells were responsible for the rapid but incomplete suppression of rotavirus shedding. Further experimentation revealed that little rotavirus antibody was made in CD4 cell-depleted BALB/c mice, and only after CD4 cells were repopulated did antibody production increase and virus shedding fully resolve. Thus, resolution of rotavirus shedding in both muMt and BALB/c mice was associated with CD4 and CD8 cell effector activities. PMID- 9343233 TI - The N-terminal side of the origin-binding domain of simian virus 40 large T antigen is involved in A/T untwisting. AB - We investigated the role of the N-terminal side of simian virus 40 (SV40) large T antigen's origin-binding domain in the initiation of virus DNA replication by analyzing the biochemical activities of mutants containing single point substitutions or deletions in this region. Four mutants with substitutions at residues between 121 and 135 were partially defective in untwisting the A/T-rich track on the late side of the origin but were normal in melting the imperfect palindrome (IP) region on the early side. Deletion of the N-terminal 109 amino acids had no effect on either activity, whereas a longer deletion, up to residue 123, greatly reduced A/T untwisting but not IP melting. These results indicate that the region from residue 121 to 135 is important for A/T untwisting but not for IP melting and demonstrate that these activities are separable. Two point substitution mutants (126PS and 135PL) were characterized further by testing them for origin DNA binding, origin unwinding, oligomerization, and helicase activity. These two mutants were completely defective in origin (form U(R)) unwinding but normal in the other activities. Our results demonstrate that a failure to normally untwist the A/T track is correlated with a defect in origin unwinding. Further, they indicate that some mutants with substitutions in the region from residue 121 to 135 interact with origin DNA incorrectly, perhaps by failing to make appropriate contacts with the A/T-rich DNA. PMID- 9343234 TI - Evolutionary variants of the human immunodeficiency virus type 1 V3 region characterized by using a heteroduplex tracking assay. AB - Syncytium-inducing (SI) variants of human immunodeficiency virus type 1 (HIV-1) are evolutionary variants that are associated with rapid CD4+ cell loss and rapid disease progression. The heteroduplex tracking assay (HTA) was used to detect evolutionary V3 variants by amplifying the V3 sequences from viral RNA derived from 50 samples of patient plasma. For this V3-specific HTA (V3-HTA), heteroduplexes were formed between the patient V3 sequences and a probe with the subtype B consensus V3 sequence. Evolution was then measured by divergence from the consensus. The presence of evolutionary variants was correlated with SI detection data on the same samples from the MT-2 cell culture assay. Evolutionary variants were correlated with the SI phenotype in 88% of the samples, and 96% of the SI samples contained evolutionary variants. In most cases the evolutionary V3 variants represented discrete clonal outgrowths of virus. Sequence analysis of the six discordant samples that did not show this correlation indicated that three non-syncytium-inducing (NSI) samples had V3 sequences that had evolved away from the consensus sequence but not toward an SI genotype. A fourth sample showed little evolution away from the consensus but was SI, which indicates that not all SI variants require basic substitutions in V3. The other two samples had SI-like genotypes and NSI phenotypes, suggesting that V3-HTA was able to detect SI emergence in these samples in the absence of their detection in vitro. V3-HTA was also used to confirm SI variant selection in MT-2 cells and to examine the possibility of variant selection during virus culture in peripheral blood cells. PMID- 9343235 TI - Poliovirus 2C region functions during encapsidation of viral RNA. AB - We have been exploring the mechanism of action of 5-(3,4-dichlorophenyl) methylhydantoin (hydantoin), an antiviral drug that inhibits the replication of poliovirus in culture. By varying the time of drug addition to infected cells, we found that the drug acts at a stage which is late in the replication cycle and subsequent to the step inhibited by guanidine. Furthermore, we detected normal levels of full-length plus-strand virion RNA in hydantoin-treated cultures. A new assembly intermediate in addition to the expected assembly intermediates was detected in drug-treated cultures. This intermediate has properties consistent with that of a packaging intermediate. Drug-resistant mutants were readily isolated. Sequence analysis of three independent drug-resistant mutants identified amino acid substitutions in the 2C coding region. Reconstruction by site-directed mutagenesis confirmed that these single mutations were sufficient to confer drug resistance. Taken together, these data suggest that the poliovirus 2C region is involved in virus encapsidation and that hydantoin inhibits this stage of replication. PMID- 9343236 TI - Dimerization of simian virus 40 T-antigen hexamers activates T-antigen DNA helicase activity. AB - Chromosomal DNA replication in higher eukaryotes takes place in DNA synthesis factories containing numerous replication forks. We explored the role of replication fork aggregation in vitro, using as a model the simian virus 40 (SV40) large tumor antigen (T antigen), essential for its DNA helicase and origin binding activities. Previous studies have shown that T antigen binds model DNA replication forks primarily as a hexamer (TAgH) and to a lesser extent as a double hexamer (TAgDH). We find that DNA unwinding in the presence of ATP or other nucleotides strongly correlates with the formation of TAgDH-DNA fork complexes. TAgH- and TAgDH-fork complexes were isolated, and the TAgDH-bound fork was denatured at a 15-fold-higher rate during the initial times of unwinding. TAgDH bound preferentially to a DNA substrate containing a 50-nucleotide bubble, indicating the bridging of each single-stranded DNA/duplex DNA junction, and this DNA molecule was also unwound at a high rate. Both the TAgH- and TAgDH-fork complexes were relatively stable, with the half-life of the TAgDH-fork complex greater than 40 min. Our data therefore indicate that the linking of two viral replication forks serves to activate DNA replication. PMID- 9343237 TI - A functional role for the conserved protonatable hairpins in the 5' untranslated region of turnip yellow mosaic virus RNA. AB - The 5' untranslated region (UTR) of the RNA of several tymoviruses contains conserved hairpins with protonatable internal loops, consisting of C-C and C-A mismatches (K. Hellendoorn, P. J. A. Michiels, R. Buitenhuis, and C. W. A. Pleij, Nucleic Acids Res. 24, 4910-4917, 1996). Here, we present a functional analysis of the 5' UTR of turnip yellow mosaic virus (TYMV) RNA, which contains two protonatable hairpins with nearly identical internal loops. Mutations were introduced in an infectious cDNA clone, and T7 RNA transcripts were used to infect Chinese cabbage plants. Different symptoms were observed for the various mutants, pointing to a functional role of the C-C and C-A mismatches in the hairpins of the 5' UTR. The replication of the virus is influenced by the mutations made, while in vitro translation studies showed that the expression of the two overlapping reading frames of TYMV is not influenced by the secondary structure of the leader. Various mutants were propagated for up to five serial passages of infection, and the sequence of the 5' UTR was determined. This resulted in virus RNA with new non-wild-type sequences that produced the wild type phenotype in infected plants. Remarkably, in all cases C-C or C-A mismatches were introduced. The internal loop of the 5'-proximal hairpin seems to be more important for the viral life cycle than that of the second hairpin. A deletion of 75% of the leader, including the two hairpins, resulted in a virus that was deficient in viral spread. Since the ratio between filled and empty capsids was changed drastically by this mutation, a role of the 5' UTR in viral packaging is proposed. PMID- 9343238 TI - Recombinant adeno-associated virus type 2 replication and packaging is entirely supported by a herpes simplex virus type 1 amplicon expressing Rep and Cap. AB - Recombinant adeno-associated virus (AAV) type 2 (rAAV) vectors have recently been shown to have great utility as gene transfer agents both in vitro and in vivo. One of the problems associated with the use of rAAV vectors has been the difficulty of large-scale vector production. Low-efficiency plasmid transfection of the rAAV vector and complementing AAV type 2 (AAV-2) functions (rep and cap) followed by superinfection with adenovirus has been the standard approach to rAAV production. The objectives of this study were to demonstrate the ability of a recombinant herpes simplex virus type 1 (HSV-1) amplicon expressing AAV-2 Rep and Cap to support replication and packaging of rAAV vectors. HSV-1 amplicon vectors were constructed which contain the AAV-2 rep and cap genes under control of their native promoters (p5, p19, and p40). An HSV-1 amplicon vector, HSV-RC/KOS or HSV RC/d27, was generated by supplying helper functions with either wild-type HSV-1 (KOS strain) or the ICP27-deleted mutant of HSV-1, d27-1, respectively. Replication of the amplicon stocks is not inhibited by the presence of AAV-2 Rep proteins, which highlights important differences between HSV-1 and adenovirus replication and the mechanism of providing helper function for productive AAV infection. Coinfection of rAAV and HSV-RC/KOS resulted in the replication and amplification of rAAV genomes. Similarly, rescue and replication of rAAV genomes occurred when rAAV vector plasmids were transfected into cells followed by HSV RC/KOS infection and when two rAAV proviral cell lines were infected with HSV RC/KOS or HSV-RC/d27. Production of infectious rAAV by rescue from two rAAV proviral cell lines has also been achieved with HSV-RC/KOS and HSV-RC/d27. The particle titer of rAAV produced with HSV-RC/d27 is equal to that achieved by supplying rep and cap by transfection followed by adenovirus superinfection. Importantly, no detectable wild-type AAV-2 is generated with this approach. These results demonstrate that an HSV-1 amplicon expressing the AAV-2 genes rep and cap along with HSV-1 helper functions supports the replication and packaging of rAAV vectors in a scaleable process. PMID- 9343239 TI - RNA aptamers specifically interact with the prion protein PrP. AB - We have isolated RNA aptamers which are directed against the recombinant Syrian golden hamster prion protein rPrP23-231 (rPrPc) fused to glutathione S transferase (GST). The aptamers did not recognize the fusion partner GST or the fusion protein GST::rPrP90-231 (rPrP27-30), which lacks 67 amino acids from the PrP N terminus. The aptamer-interacting region of PrPc was mapped to the N terminal amino acids 23 to 52. Sequence analyses suggest that the RNA aptamers may fold into G-quartet-containing structural elements. Replacement of the G residues in the G quartet scaffold with uridine residues destroyed binding to PrP completely, strongly suggesting that the G quartet motif is essential for PrP recognition. Individual RNA aptamers interact specifically with prion protein in brain homogenates from wild-type mice (C57BL/6), hamsters (Syrian golden), and cattle as shown by supershifts obtained in the presence of anti-PrP antibodies. No interaction was observed with brain homogenates from PrP knockout mice (prn p(0/0)). Specificity of the aptamer-PrP interaction was further confirmed by binding assays with antisense aptamer RNA or a mutant aptamer in which the guanosine residues in the G tetrad scaffold were replaced by uridine residues. The aptamers did not recognize PrP27-30 in brain homogenates from scrapie infected mice. RNA aptamers may provide a first milestone in the development of a diagnostic assay for the detection of transmissible spongiform encephalopathies. PMID- 9343240 TI - The role of Kupffer cell activation and viral gene expression in early liver toxicity after infusion of recombinant adenovirus vectors. AB - Systemic application of first-generation adenovirus induces pathogenic effects in the liver. To begin unraveling the mechanisms underlying early liver toxicity after adenovirus infusion, particularly the role of macrophage activation and expression of viral genes in transduced target cells, first-generation adenovirus or adenovirus vectors that lacked most early and late gene expression were administered to C3H/HeJ mice after transient depletion of Kupffer cells by gadolinium chloride treatment. Activation of NF-kappaB, and the serum levels of the proinflammatory cytokines tumor necrosis factor (TNF) and interleukin-6 (IL 6) were studied in correlation with liver damage, apoptosis, and hepatocellular DNA synthesis. While Kupffer cell depletion nearly eliminated adenovirus-induced TNF release, it resulted in a more robust IL-6 release. These responses were greatly reduced in animals receiving the deleted adenovirus. Although there were quantitative differences, NF-kappaB activation was observed within minutes of first-generation or deleted adenovirus vector administration regardless of the status of the Kupffer cells, suggesting that the induction is related to a direct effect of the virus particle on the hepatocyte. Early liver toxicity as determined by serum glutamic-pyruvic transaminase elevation and inflammatory cell infiltrates appeared to be dependent on adenovirus-mediated early gene expression and intact Kupffer cell function. Kupffer cell depletion had little effect on adenovirus-mediated hepatocyte apoptosis but did increase hepatocellular DNA synthesis. Finally, Kupffer cell depletion decreased the persistence of transgene (human alpha1-antitrypsin [hAAT]) expression that was associated with a more pronounced humoral immune response against hAAT. The elucidation of these events occurring after intravenous adenovirus injection will be important in developing new vectors and transfer techniques with reduced toxicity. PMID- 9343241 TI - Structure-based identification of an inducer of the low-pH conformational change in the influenza virus hemagglutinin: irreversible inhibition of infectivity. AB - Past efforts to employ a structure-based approach to design an inhibitor of the fusion-inducing conformational change in the influenza virus hemagglutinin (HA) yielded a family of small benzoquinones and hydroquinones. The most potent of these, tert-butyl hydroquinone (TBHQ), inhibits both the conformational change in HA from strain X:31 influenza virus and viral infectivity in tissue culture cells with 50% inhibitory concentrations in the micromolar range (D. L. Bodian, R. B. Yamasaki, R. L. Buswell, J. F. Stearns, J. M. White, and I. D. Kuntz, Biochemistry 32:2967-2978, 1993). A new structure-based inhibitor design search was begun which involved (i) the recently refined crystal structure (2.1-A resolution) of the HA ectodomain, (ii) new insights into the conformational change, and (iii) improvements in the molecular docking program, DOCK. As a result, we identified new inhibitors of HA-mediated membrane fusion. Like TBHQ, most of these molecules inhibit the conformational change. One of the new compounds, however, facilitates rather than inhibits the HA conformational change. Nonetheless, the facilitator, diiodofluorescein, inhibits HA-mediated membrane fusion and, irreversibly, infectivity. We further characterized the effects of inhibitors from both searches on the conformational change and membrane fusion activity of HA as well as on viral infectivity. We also isolated and characterized several mutants resistant to each class of inhibitor. The implications of our results for HA-mediated membrane fusion, anti-influenza virus therapy, and structure-based inhibitor design are discussed. PMID- 9343242 TI - A hypothalamic neuronal cell line persistently infected with scrapie prions exhibits apoptosis. AB - Neuronal death and vacuolation are characteristics of the CNS degeneration found in prion diseases. Relatively few cultured cell lines have been identified that can be persistently infected with scrapie prions, and none of these cells show cytopathologic changes reminiscent of prion neuropathology. The differentiated neuronal cell line GT1, established from gonadotropin hormone releasing-hormone neurons immortalized by genetically targeted tumorigenesis in transgenic mice (P. L. Mellon, JJ. Windle, P. C. Goldsmith, C. A. Padula, J. L. Roberts, and R. I. Weiner, Neuron 5:1-10, 1990), was examined for its ability to support prion formation. We found that GT1 cells could be persistently infected with mouse RML prions and that conditioned medium from infected cells could transfer prions to uninfected cells. In many but not all experiments, a subpopulation of cells showed reduced viability, morphological signs of neurodegeneration and vacuolation, and features of apoptosis. Subclones of GT1 cells that were stably transfected with the trk4 gene encoding the high-affinity nerve growth factor (NGF) receptor (GT1-trk) could also be persistently infected. NGF increased the viability of the scrapie-infected GT1-trk cells and reduced the morphological and biochemical signs of vacuolation and apoptosis. GT1 cells represent a novel system for studying the molecular mechanisms underlying prion infectivity and subsequent neurodegenerative changes. PMID- 9343243 TI - Transcription activities of human papillomavirus type 11 E6 promoter-proximal elements in raft and submerged cultures of foreskin keratinocytes. AB - Human papillomaviruses (HPVs) replicate only in differentiated squamous epithelia in warts and in epithelial raft cultures grown at the medium-air interface. Virus encoded and host transcription factors are thought to be responsible for repressing the viral enhancer and promoter located within the upstream regulatory region (URR) in the undifferentiated basal and parabasal cells while up regulating their activities in the differentiated spinous cells. Using recombinant retroviruses, we acutely transduced neonatal foreskin keratinocytes (PHKs) with a lacZ reporter gene driven by the wild-type URR of the low-risk HPV type 11 or by a URR with individual mutations in seven promoter-proximal elements, some of which have not been characterized previously. Beta galactosidase activities were detected in the submerged, proliferating PHKs and also in the differentiated spinous cells, but not in the steady-state proliferating basal cells, of stratified raft cultures. In particular, mutation of an Oct1, an Sp1, or a previously unknown promoter-proximal AP1 site severely reduced the reporter activity, whereas mutation of either of two NF1 sites flanking the Oct1 site had no effect. These results demonstrate changes in cellular transcription factor profiles under different culture conditions and begin to characterize the naturally differentiation-dependent activation of the URR. They provide one molecular explanation for the patterns of HPV expression in warts and help validate epithelial raft cultures as an important experimental system for genetic dissection of HPV regulatory elements. PMID- 9343244 TI - Complement component 3 interactions with coxsackievirus B3 capsid proteins: innate immunity and the rapid formation of splenic antiviral germinal centers. AB - Innate immunity is central to the clearance of pathogens from hosts as well as to the definition of acquired immune responses (D. T. Fearon, and R. M. Locksley, Science 272:50-53, 1996). Coxsackievirus B3 (CVB3), a human cardiopathic virus, was evaluated for the ability to activate the alternative and classical pathway of complement. CVB3 proteins interact with complement component 3 (C3, a soluble protein effector of innate immunity) after either in vitro exposure to mouse serum or in vivo murine infection and activate the alternative pathway of complement. In addition, we demonstrate that viral antigen retention and localization in germinal centers is dependent on C3, while virus antigen retention in extrafollicular regions in the spleen is not. In vivo depletion of native C3 abolished the rapid formation of virus-specific germinal centers (by day 3 post-CVB3 infection) in the absence of serum anti-CVB3 antibodies. These studies demonstrate that innate immune mechanisms, such as C3 interaction with CVB3, are essential for splenic antiviral germinal center formation in naive (antigen nonsensitized) mice resistant (C57BL/6J strain) and susceptible (A/J strain) to CVB3-induced myocarditis. PMID- 9343245 TI - Attenuated replication of human immunodeficiency virus type 1 with a didanosine selected reverse transcriptase mutation. AB - The Leu-74 to Val (Leu74Val) mutation in human immunodeficiency virus type 1 reverse transcriptase (RT) develops as a consequence of didanosine (ddI) therapy and is associated with a decreased susceptibility to ddI. In this report, we provide evidence that the ddI-associated Leu74Val mutation confers a replication disadvantage to the virus. In a series of experiments, we have shown that (i) a cloned virus with an engineered Leu74Val mutation in RT was attenuated for replication; (ii) a Val-to-Leu revertant of Leu74Val in the pNL4-3 background replicated with an efficiency similar to that of the wild-type virus; (iii) when two isolates from the same patient were compared, a clinical isolate containing mutations Leu74Val and Thr215Tyr was attenuated for replication compared to one in which the Thr215Tyr mutation alone was present; and (iv) the viruses with the Leu74Val mutation showed an 11% loss of fitness in a single passage compared to the wild-type and a mutant virus containing a Lys70Arg mutation. The loss of fitness for viruses grown in drug-free medium could result in an inability to detect a Leu74Val mutant in clinical isolates obtained post-ddI therapy. The decreased replication ability of the Leu74Val mutant virus selected by ddI therapy provides a strong rationale for the lower viral RNA levels observed with ddI therapy compared to zidovudine therapy in clinical trials. PMID- 9343246 TI - Unique double-stranded RNAs responsible for the anti-Candida activity of the yeast Hanseniaspora uvarum. AB - Killer strains of the yeast Hanseniaspora uvarum contain cytoplasmic double stranded RNAs (dsRNAs) of 4.7-kbp L and 1.0-kbp M species, which were shown to be separately packaged into icosahedral virus-like particles exhibiting RNA dependent RNA polymerase activity. The L genome of the H. uvarum L-dsRNA virion HuV-L was shown to encode a 77-kDa major capsid protein. Peptide maps of the purified HuV coat protein and the 81-kDa major capsid protein from K1 killer viruses of Saccharomyces cerevisiae revealed distinctly different peptide patterns, suggesting significant sequence divergence at the level of the capsid coding L-dsRNAs. In vitro transcripts from purified HuV-L particles showed no cross-hybridization to denatured L(A), L(B), or L(C), indicating that L from H. uvarum represents a unique L-dsRNA species. Weak, but clearly detectable cross hybridization of the 1.0-kb dsRNA of HuV-M, encoding the secreted 18-kDa anti Candida toxin, to the toxin-coding M genomes of S. cerevisiae K1, K2, and K28 killers indicated partial sequence homology among all of the M-dsRNAs tested. PMID- 9343247 TI - Hepatitis C virus nonstructural region 5A protein is a potent transcriptional activator. AB - The hepatitis C virus (HCV) nonstructural region 5A (NS5A) protein, without its 146 amino-terminal amino acids and fused to the DNA-binding domain of GAL4, strongly activates transcription in yeast and human hepatoma cells. Transcriptional activation by the HCV NS5A protein may play a role in viral replication and hepatocarcinogenesis. PMID- 9343248 TI - Mouse susceptibility to mouse hepatitis virus infection is linked to viral receptor genotype. AB - We have reported that the receptor for mouse hepatitis virus (MHV) expressed in MHV-susceptible BALB/c mice (MHVR1) has 10 to 30 times the virus-binding activity of the MHV receptor expressed in MHV-resistant SJL mice (MHVR2) (N. Ohtsuka, Y. K. Yamada, and F. Taguchi, J. Gen. Virol. 77:1683-1992, 1996). This fact indicates the possibility that the difference in MHV susceptibility between BALB/c and SJL mice is determined by the virus-binding activity of the receptor. To test this possibility, we have examined MHV susceptibility in mice with the homozygous MHVR1 gene (R1/R1 genotype), mice with the MHVR1 and MHVR2 genes (R1/R2 genotype), and mice with the homozygous MHVR2 gene (R2/R2 genotype) produced by cross and backcross mating between BALB/c and SJL mice. All 63 F2 and backcrossed mice with the MHVR1 gene (R1/R1 and R1/R2) were susceptible to MHV infection, and all 57 with the homozygous MHVR2 gene (R2/R2) were resistant. We have also examined the MHV receptor genotypes of several mouse strains that were reported to be susceptible to MHV infection. All of those mice had the MHVR1 gene. These results suggest the possibility that the viral receptor determines the susceptibility of the whole animal to MHV infection. PMID- 9343249 TI - Infection of macrophages by Theiler's murine encephalomyelitis virus is highly dependent on their activation or differentiation state. AB - Macrophages are the main targets of Theiler's murine encephalomyelitis virus (TMEV) during persistent demyelinating infection of mice. Replication of TMEV in macrophages was previously shown to depend on their activation state. Here, we show that the quality of the serum used for culture drastically influences viral entry in RAW264.7 macrophages. PMID- 9343250 TI - Replication-competent picornaviruses with complete genomic RNA 3' noncoding region deletions. AB - The genomic RNA 3' noncoding region is believed to be a major cis-acting molecular genetic determinant for regulating picornavirus negative-strand RNA synthesis by promoting replication complex recognition. We report the replication of two picornavirus RNAs harboring complete deletions of the genomic RNA 3' noncoding regions. Our results suggest that while specific 3'-terminal RNA sequences and/or secondary structures may have evolved to promote or regulate negative-strand RNA synthesis, the basic mechanism of replication initiation is not strictly template specific and may rely primarily upon the proximity of newly translated viral replication proteins to the 3' terminus of template RNAs within tight membranous replication complexes. PMID- 9343251 TI - An early, abundant cytotoxic T-lymphocyte response against Theiler's virus is critical for preventing viral persistence. AB - In genetically susceptible strains of mice, the DA strain of Theiler's virus, a picornavirus, causes a persistent infection of the white matter of the spinal cord associated with chronic demyelination. In resistant strains, on the other hand, the infection is cleared within 1 to 2 weeks. In this article, we show that Theiler's virus induces a rapid and abundant cytotoxic T lymphocyte (CTL) response in resistant C57BL/6 mice, while the response remains low throughout infection in susceptible SJL/J mice. This difference can be referred to a higher number of virus-specific CTL precursors in C57BL/6 mice. These observations indicate that the efficient induction of virus-specific CTL precursors is critical for avoiding the establishment of a persistent picornaviral infection. PMID- 9343252 TI - A second form of infectious bursal disease virus-associated tubule contains VP4. AB - Preparations of density gradient-purified infectious bursal disease virus (IBDV) were found to contain full and empty icosahedral virions, type I tubules with a diameter of about 60 nm, and type II tubules 24 to 26 nm in diameter. By immunoelectron microscopy we demonstrate that virions and both types of tubular structures specifically react with anti-IBDV serum. In infected cells intracytoplasmic and intranuclear type II tubules reacted exclusively with an anti-VP4 monoclonal antibody, as did type II tubules in virion preparations. The immunofluorescence pattern with the anti-VP4 antibody correlated with electron microscopical findings. Neither purified extracellular nor intracellular virions were labeled with the anti-VP4 MAb. Our data show that the type II tubules contain VP4 and suggest that VP4 is not part of the virus particle. PMID- 9343255 TI - Details of the arrangement of the outer capsid of rice dwarf phytoreovirus, as visualized by two-dimensional crystallography. AB - Two-dimensional crystals were obtained from purified P8, an outer capsid protein of rice dwarf phytoreovirus. A filtered image of the two-dimensional crystal, in combination with the results of biochemical analysis, revealed the unit formation of the capsid protein, a capsomere structure, which appeared to be an approximately equilateral triangle with sides of approximately 6 nm and which was composed of a trimer of P8 protein. Details of the arrangements of the outer capsid of the virus are described. PMID- 9343253 TI - Functional complementation of UL3.5-negative pseudorabies virus by the bovine herpesvirus 1 UL3.5 homolog. AB - The UL3.5 gene is positionally conserved but highly variable in size and sequence in different members of the Alphaherpesvirinae and is absent from herpes simplex virus genomes. We have shown previously that the pseudorabies virus (PrV) UL3.5 gene encodes a nonstructural protein which is required for secondary envelopment of intracytoplasmic virus particles in the trans-Golgi region. In the absence of UL3.5 protein, naked nucleocapsids accumulate in the cytoplasm, release of infectious virions is drastically reduced, and plaque formation in cell culture is inhibited (W. Fuchs, B. G. Klupp, H. Granzow, H.-J. Rziha, and T. C. Mettenleiter, J. Virol. 70:3517-3527, 1996). To assay functional complementation by a heterologous herpesviral UL3.5 protein, the UL3.5 gene of bovine herpesvirus 1 (BHV-1) was inserted at two different sites within the genome of UL3.5-negative PrV. In cells infected with the PrV recombinants the BHV-1 UL3.5 gene product was identified as a 17-kDa protein which was identical in size to the UL3.5 protein detected in BHV-1-infected cells. Expression of BHV-1 UL3.5 compensated for the lack of PrV UL3.5, resulting in a ca. 1,000-fold increase in virus titer and restoration of plaque formation in cell culture. Also, the intracellular block in viral egress was resolved by the BHV-1 UL3.5 gene. We conclude that the UL3.5 proteins of PrV and BHV-1 are functionally related and are involved in a common step in the egress of alphaherpesviruses. PMID- 9343254 TI - Susceptibility of human immunodeficiency virus type 1 group O isolates to antiretroviral agents: in vitro phenotypic and genotypic analyses. AB - We investigated the phenotypic and genotypic susceptibility of 11 human immunodeficiency virus type 1 (HIV-1) group O strains to nucleoside and nonnucleoside reverse transcriptase (RT) inhibitors and protease inhibitors in vitro. Phenotypic susceptibility was determined by using a standardized in vitro assay of RT inhibition, taking into account the replication kinetics of each strain. HIV-1 group M and HIV-2 isolates were used as references. DNA from cocultured peripheral blood mononuclear cells was amplified by using pol-specific group O primers and cloned for sequencing. Group O isolates were highly sensitive to nucleoside inhibitors, but six isolates were naturally highly resistant to all of the nonnucleoside RT inhibitors tested. Phylogenetic analysis of the pol gene showed that these isolates formed a separate cluster within group O, and genotypic analysis revealed a tyrosine-to-cysteine substitution at residue 181. Differences in susceptibility to saquinavir and ritonavir (RTV) were not significant between group O and group M isolates, although the 50% inhibitory concentration of RTV for group O isolates was higher than that for the HIV-1 subtype B strains. The study of HIV-1 group O susceptibility to antiretroviral drugs revealed that the viruses tested had specific phenotypic characteristics contrasting with the group M phenotypic expression. PMID- 9343256 TI - Persistence of recombinant adenovirus in vivo is not dependent on vector DNA replication. AB - Recombinant adenovirus vectors represent an efficient means of transferring genes into many different organs. The first-generation E1-deleted vector genome remains episomal and, in the absence of host immunity, persists long-term in quiescent tissues such as the liver. The mechanism(s) which allows for persistence has not been established; however, vector DNA replication may be important because replication has been shown to occur in tissue culture systems. We have utilized a site-specific methylation strategy to monitor the replicative fate of E1-deleted adenovirus vectors in vitro and in vivo. Methylation-marked adenovirus vectors were produced by the addition of a methyl group onto the N6 position of the adenine base of XhoI sites, CTCGAG, by propagation of vectors in 293 cells expressing the XhoI isoschizomer PaeR7 methyltransferase. The methylation did not affect vector production or transgene expression but did prevent cleavage by XhoI. Loss of methylation through viral replication restores XhoI cleavage and was observed by Southern analysis in a wide variety of, but not all, cell culture systems studied, including hepatoma and mouse and macaque primary hepatocyte cultures. In contrast, following liver-directed gene transfer of methylated vector in C57BL/6 mice, adenovirus vector DNA was not cleaved by XhoI and therefore did not replicate, even after a period of 3 weeks. Although replication may occur in some tissues, these results show that stabilization of the vector within the target tissue prior to clearance by host immunity is not dependent upon replication of the vector, demonstrating that the input transduced DNA genomes were the persistent molecules. This information will be useful for the design of optimal adenovirus vectors and perhaps nonviral episomal vectors for clinical gene therapy. PMID- 9343257 TI - Cross-clade human immunodeficiency virus (HIV)-specific cytotoxic T-lymphocyte responses in HIV-infected Zambians. AB - We have examined cross-clade HIV-specific cytotoxic T-lymphocyte (CTL) activity in peripheral blood of eight Zambian individuals infected with non-B-clade human immunodeficiency virus type 1 (HIV-1). Heteroduplex mobility assay and partial sequence analysis of env and gag genes strongly suggests that all the HIV infected subjects were infected with clade C HIV-1. Six of eight C-clade HIV infected individuals elicited CTL activity specific for recombinant vaccinia virus-infected autologous targets expressing HIV gag-pol-env derived from B-clade HIV-1 (IIIB). Recognition of individual recombinant HIV-1 B-clade vaccinia virus infected targets expressing gag, pol, or env was variable among the patients tested, indicating that cross-clade CTL activity is not limited to a single HIV protein. These data demonstrate that HIV clade C-infected individuals can mount vigorous HIV clade B-reactive CTL responses. PMID- 9343258 TI - Mutational analysis of human T-cell leukemia virus type 2 Tax. AB - A mutational analysis of human T-cell leukemia virus type 2 (HTLV-2) Tax (Tax-2) was performed to identify regions within Tax-2 important for activation of promoters through the CREB/ATF or NF-kappaB/Rel signaling pathway. Tax-2 mutations within the putative zinc-binding region as well as mutations at the carboxy terminus disrupted CREB/ATF transactivation. A single mutation within the central proline-rich region of Tax-2 disrupted the transactivation of the NF kappaB/Rel pathway. Surprisingly, this mutation, which is thought to be in a separate activation domain, was suppressed by mutations within or around the putative zinc-binding region, suggesting an interaction between these two regions. These analyses indicate that the functional regions or domains important for transactivation through the CREB/ATF or NF-kappaB/Rel signaling pathway are similar, but not identical, in Tax-1 and Tax-2. Identification of these distinct Tax-2 mutants should facilitate comparative biological studies of HTLV-1 and HTLV 2 and ultimately lead to the determination of the functional importance of Tax trans-acting capacities in T-lymphocyte transformation by HTLV. PMID- 9343260 TI - A small region of the ecotropic murine leukemia virus (MuLV) gag gene profoundly influences the types of polytropic MuLVs generated in mice. AB - The vast majority of recombinant polytropic murine leukemia viruses (MuLVs) generated in mice after infection by ecotropic MuLVs can be classified into two major antigenic groups based on their reactivities to two monoclonal antibodies (MAbs) termed Hy 7 and 516. These groups very likely correspond to viruses formed by recombination of the ecotropic MuLV with two distinct sets of polytropic env genes present in the genomes of inbred mouse strains. We have found that nearly all polytropic MuLVs identified in mice infected with a substrain of Friend MuLV (F-MuLV57) are reactive with Hy 7, whereas mice infected with Moloney MuLV (Mo MuLV) generate major populations of both Hy 7- and 516-reactive polytropic MuLVs. We examined polytropic MuLVs generated in NFS/N mice after inoculation with Mo MuLV-F-MuLV57 chimeras to determine which regions of the viral genome influence this difference between the two ecotropic MuLVs. These studies identified a region of the MuLV genome which encodes the nucleocapsid protein and a portion of the viral protease as the only region that influenced the difference in polytropic-MuLV generation by Mo-MuLV and F-MuLV57. PMID- 9343259 TI - Human immunodeficiency virus type 2 envelope glycoprotein binds to CD8 as well as to CD4 molecules on human T cells. AB - We report here that human immunodeficiency virus type 2 (HIV-2) envelope glycoprotein (gp105), but not HIV-1 gp120, can bind to CD8 molecules as well as to CD4 molecules on human T cells. This phenomenon may lead to differences in the life cycles of HIV-1 and HIV-2, and it may be related to the differences in disease manifestations of HIV-1 and HIV-2 infection, including longer survival of HIV-2-infected patients. PMID- 9343261 TI - Bovine herpesvirus 4 BORFE2 protein inhibits Fas- and tumor necrosis factor receptor 1-induced apoptosis and contains death effector domains shared with other gamma-2 herpesviruses. AB - Fas- and tumor necrosis factor receptor 1 (TNFR1)-induced apoptosis is mediated by the interaction of FADD with caspase-8. Here, we report that the bovine herpesvirus 4 (BHV4) BORFE2 gene encodes a protein that inhibits Fas- and TNFR1 induced apoptosis and contains death effector domains (DEDs). Using the yeast two hybrid system, we found that the BORFE2 protein interacts with the prodomain of caspase-8. Furthermore, we show that BHV4 BORFE2 is a member of a family of DED containing proteins that includes other gamma-2 herpesviruses, such as Kaposi's sarcoma-associated herpesvirus and herpesvirus saimiri. PMID- 9343262 TI - Export of hepatitis B virus RNA on a Rev-like pathway: inhibition by the regenerating liver inhibitory factor IkappaB alpha. AB - Nuclear export of hepatitis B virus (HBV) RNA is mediated by a specific RNA element but, in contrast to lentivirus genomic RNA, does not depend on viral proteins. We show that nonetheless, the export of HBV RNA can be blocked by competitive inhibitors of Rev-mediated lentivirus RNA export, suggesting that the export pathways of both viral species share components and might be driven by the same nuclear export machinery. HBV RNA export is also inhibited by overexpression of IkappaB alpha, as reported previously for the export of human immunodeficiency virus RNA. Since IkappaB alpha is strongly overexpressed during liver regeneration, its inhibition of HBV RNA export might contribute to elimination or silent persistence of HBV. PMID- 9343263 TI - Implication of a cis-acting element in the cytoplasmic accumulation of unspliced Borna disease virus RNAs. AB - Borna disease virus (BDV), the prototype of a new family within the order Mononegavirales, is unusual in its nuclear localization for replication and transcription and use of RNA splicing for gene expression. The BDV antigenome contains three transcription units and six major open reading frames. Multicistronic RNAs containing two introns are elaborated from the third transcription unit. Differential splicing of the two introns and cytoplasmic accumulation of the unspliced and partially spliced RNA are critical for the balanced expression of the putative matrix protein, glycoprotein, and polymerase. To investigate the mechanisms for cytoplasmic expression of unspliced and partially spliced BDV transcripts, the levels of these transcripts were measured in the cytoplasm of infected COS-7 cells and noninfected COS-7 cells transfected with plasmids containing 2.8-kb cDNA inserts representing either wild-type or mutant BDV RNA from the third transcription unit. Analysis of truncation mutations allowed the identification of a cis-acting element present within the 3' end of the BDV 2.8-kb transcript that facilitated the cytoplasmic accumulation of unspliced BDV transcripts through nucleocytoplasmic transport. The nucleocytoplasmic transport activity was not dependent on the presence of BDV proteins. Gel-shift assays revealed that the cis-acting element binds specifically to host cytoplasmic and nuclear proteins. PMID- 9343264 TI - Construction of an adenovirus type 7a E1A- vector. AB - A strategy for constructing replication-defective adenovirus vectors from non subgroup C viruses has been successfully demonstrated with adenovirus type 7 strain a (Ad7a) as the prototype. An E1A-deleted Ad7a reporter virus expressing the chloramphenicol acetyltransferase (CAT) gene from the cytomegalovirus promoter enhancer was constructed with DNA fragments isolated from Ad7a, an Ad7a recombination reporter plasmid, and the 293 cell line. The Ad7a-CAT virus particle transduces A549 cells as efficiently as Ad5-based vectors. Intravenous infections in a murine model indicate that the Ad7a-CAT virus infects a variety of tissues, with maximal levels of CAT gene expression found in the liver. The duration of Ad7a-CAT transgene expression in the liver was maximally maintained 2 weeks postinfection, with a decline to baseline activity by the week 4 postinfection. Ad7a-CAT represents the first example of a non-subgroup C E1A- adenovirus gene transfer vector. PMID- 9343265 TI - Ultrasonography screening for developmental dysplasia of the hip joint. AB - The use of ultrasonography as a complement to clinical examination will increase the reliability of the evaluation of unstable hips in newborn infants. In particular, the number of false-positive Ortolani and Barlow tests will decrease. However, the interpretation of the ultrasonogram in newborn infants has a steep learning curve with considerable risk of a high number of false-positive hips being diagnosed. Therefore, universal screening for developmental dysplasia of the hip by ultrasonography cannot yet be recommended from a cost-benefit point of view. PMID- 9343266 TI - The growing allergy problem. PMID- 9343267 TI - Pathophysiology and impact of nocturnal enuresis. AB - Nocturnal enuresis in children is not a psychogenic disorder. It is caused by a hereditary delay in maturation of the somatic mechanisms (reduction of nocturnal urine production and a normal arousal to a full bladder) which prevent the child from wetting the bed. Traditionally, doctors treating bedwetting children have used an expectant attitude, because nocturnal enuresis has been looked upon as self-limiting and harmless. According to recent research this is not true. More than 5% of children and 0.5% of the adult population report nocturnal enuresis, meaning that 10% of enuretic children will remain bedwetters for life if left untreated, and nocturnal enuresis is perceived as a shameful condition, giving a significant impairment of self-esteem at an age when an intact self-image is extremely important for an optimal development of the child's personality. Treatment should be given when the enuretic child wants to sleep dry. PMID- 9343268 TI - Malnutrition, cell-mediated immune deficiency and acute upper respiratory infections in rural Bangladeshi children. AB - A community-based longitudinal study conducted in rural Bangladesh investigated the association between nutritional status, cell-mediated immune status and acute upper respiratory infections (URI). A total of 696 children aged 0-59 months was followed prospectively for 1 y yielding 183,865 child-days' observation. Trained field workers visited each child every 4th d and collected morbidity data on symptoms suggesting URI (cough, fever, nasal discharge) for the preceding 3 d by recall. On the day of visit they examined each child reporting cough and/or fever to record the temperature, presence of nasal discharge, rate of respiration and presence of chest indrawing. Anthropometry for all children was conducted monthly. Cell-mediated immune competence was assessed by a multiple antigen skin test at baseline and thereafter every 3 months. The incidence of URI was 5.3 episodes per child-year observed. Approximately three-quarters of the study children were below -2 Z-score weight for age and height for age, and a quarter below -2 Z-score weight for height. During different test periods 9-21% of the study children did not respond to any of the test antigens. In a regression model children < -2 Z-score for weight for height had 16% [odds ratio (OR) 1.16, 95% confidence interval (CI) 1.03-1.31, p = 0.01] higher risk of developing URI. Anergic children had 20% higher risk (OR 1.20, CI 1.05-1.38, p = 0.009) of URI than immunocompetent children. The study demonstrated that wasting and depressed cell-mediated immunity (CMI), but not stunting, were associated with the incidence of URI among rural Bangladeshi children. PMID- 9343269 TI - Increased mucosal inflammatory cytokines in children with Helicobacter pylori associated gastritis. AB - The concentrations of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL 6) and IL-1-beta in tissue homogenates of gastric mucosal biopsy specimens, and in gastric juice samples from Helicobacter pylori-positive and -negative children, were determined. The study population comprised 30 children with recurrent abdominal pain attending upper gastrointestinal endoscopy. Of these patients 18 were infected with H. pylori. Cytokine concentrations in gastric biopsy homogenate supernatants and in gastric juice were measured by enzyme linked immunosorbent assay (ELISA). TNF-alpha levels in gastric juice and in gastric biopsy homogenate supernatants in patients with H. pylori-positive gastritis were found to be significantly higher than those in children without H. pylori infection. IL-6 levels were also higher in H. pylori-infected subjects, but the difference in IL-6 concentrations measured in gastric juice and biopsy homogenate supernatants did not reach statistical significance. IL-1-beta concentrations in both specimens showed no significant difference between the two groups of children. It was suggested that increased levels of inflammatory cytokines, especially TNF-alpha and IL-6 generated locally within the gastric mucosa might be implicated in the pathogenesis of H. pylori-associated gastritis in childhood. PMID- 9343270 TI - Inhaled budesonide reduces lung hyperinflation in children with asthma. AB - Previous studies have shown that asthmatics have hyperinflation as defined by larger total lung capacity. The present study was set up in order to document changes in asthma clinic, airway calibre, airway reactivity and lung volumes after budesonide treatment. After a 2-week run-in period, 28 children with moderate persistent asthma were treated in a double-blind manner either with budesonide (0.4 mg/day) or placebo for 8 weeks and, thereafter, all patients were treated with open-label budesonide for a further 20 weeks. Symptoms, bronchodilator requirements and airway calibre improved significantly after 8 weeks of treatment (p < 0.05 for each) and prolonged treatment did not cause any further improvement. Reduction in hyperreactivity was apparent only after 20-28 weeks of treatment. Total lung capacity decreased along with budesonide treatment in both groups suggesting that early introduction of an inhaled corticosteroid may be useful in the prevention of asthma-related remodelling of the lung and thoracic cage. PMID- 9343271 TI - Reference values for height, height velocity and weight in Turner's syndrome. Swedish Study Group for GH treatment. AB - As Northern Europeans are currently the tallest people in the world, specific growth charts for girls with Turner's Syndrome from this area are needed. Based on height and weight measurements from 598 girls with Turner's Syndrome (372 from the Netherlands, 108 from Denmark, 118 from Sweden) not treated with growth promoting substances and without signs of spontaneous puberty, we constructed growth charts for height-for-age, height-velocity-for-age, weight-for-age, weight for-height and Body Mass Index for age. Reference tables and regression equations for mean and standard deviation are provided allowing calculation of Standard Deviation Scores. The height and height velocity curves show a low birth length, gradual deviation from the normal percentile curves without pubertal growth spurt, and a prolonged growth until the early 20s. Mean adult height was 146.9 +/ 7.8 cm. Mean weight-for-age was lower than in normal reference children but height-adjusted weight was higher, except in infancy and early childhood. Further studies are required on the factors influencing the weight-height relationship in Turner's Syndrome. PMID- 9343272 TI - Growth hormone therapy in pre-pubertal children with Noonan syndrome: first year growth response and comparison with Turner syndrome. AB - We studied the growth-promoting effect of treatment with recombinant human growth hormone in 23 pre-pubertal children with Noonan syndrome, aged between 5.4 and 14.3 y, and all with a height < 1.4 SD for Tanner standards. The growth response and skeletal maturation after 1 y of recombinant human growth hormone treatment (0.15 U/kg/day given by daily injection) in the Noonan syndrome patients was compared with the auxological changes observed in a group of 17 girls with Turner syndrome with a comparable age and height deficit who were treated with recombinant human growth hormone in a similar way. During 1 y of treatment, the mean +/- SD height velocity increased by 4.0 +/- 1.6 cm/y in the Noonan syndrome group and by 3.6 +/- 1.3 cm/y in the Turner syndrome group. Height SDS for chronological age in the Noonan syndrome group increased by 0.53 +/- 0.46 (p < 0.001). In the Noonan syndrome patients the changes in height velocity were positively related to birthweight (r = 0.48, p < 0.05). The changes in height velocity or height SDS were not related to the age, height deficit or a delay in bone age maturation at start of treatment. In neither the patients with Noonan syndrome nor Turner syndrome was an acceleration of bone maturation found. We conclude that treatment with recombinant human growth hormone in pre-pubertal NS patients induces an increase in height velocity and height SDS comparable to that observed in Turner syndrome girls. PMID- 9343273 TI - Marfan syndrome and other systemic disorders with congenital ectopia lentis. A Danish national survey. AB - In order to elucidate demographic and nosologic characteristics of Marfan syndrome (MS) and other systemic disorders associated with congenital ectopia lentis (ECL) in Denmark, a register of affected persons was established in a nationwide retrospective study. Three hundred and forty patients with MS (180M, 160F) were registered. By January 1, 1993, the estimated prevalence rate of MS was 4.6/100,000. An estimated average birth rate of 0.96/10,000 live born was found. The mean age at death was decreased (38 +/- 18 y) mainly due to cardiovascular complications. Correspondingly, a median cumulative probability of survival of 57 y (males) and 58 y (females) was found. Only seven patients with ECL as manifestation of other systemic disorders were diagnosed in the material, i.e. three patients with homocystinuria, two patients with Weill-Marchesani syndrome, and another 2 patients with sulphite oxidase deficiency. In a further 122 cases with ECL a nosologic diagnosis could not be established due to unavailable or insufficient information. This group may comprise patients with MS and other systemic disorders as well as patients with non-systemic ECL. In retrospect, the differential diagnostic activity for patients with congenital ectopia lentis has been insufficient during the last few decades and a common protocol for diagnosis, control and treatment of ECL is needed. PMID- 9343274 TI - Membranous duodenal stenosis. AB - The experience of our 16 patients treated for membranous duodenal stenosis is reported. Their treatment and course was analysed in a retrospective study. Eight patients were operated on within the first 16 days of life and in the remaining group surgery was performed at 1 month to 4 y of age. The presenting symptom leading to diagnosis was, in all but one case, non-bile-stained vomiting. Associated malformations were found in all but four patients, mostly morbus Down. The operative procedure performed was a partial excision of the duodenal membrane and a duodenoplasty in 10 patients, a duodenojejunostomy in 5 patients, and a duodenoplasty only in 1 patient. The postoperative course was without lethal complications. One late stricture in an anastomosis occurred. We conclude that in its presentation, duodenal stenosis differs from duodenal atresia, and can often be misinterpreted, resulting in a late diagnosis, and should be reported as a separate entity. PMID- 9343275 TI - Intestinal microflora of Estonian and Swedish infants. AB - The intestinal microflora of 1-y-old healthy Estonian (n = 27) and Swedish infants (n = 29) was studied by quantitative culture of faecal samples. The major differences were high counts of lactobacilli and eubacteria in the former and increased numbers of clostridia in the latter babies. Bifidobacteria and anaerobic cocci prevailed equally in both groups, while eubacteria and enterococci were the major microorganisms in many Estonian infants and bacteroides and clostridia in many Swedish infants. The microflora of the Estonian infants was in many aspects similar to the flora prevailing in infants of western Europe in the 1960s. The results suggest a shift in the intestinal microflora among infants in western industrialized countries. PMID- 9343276 TI - The decline in the incidence of SIDS in Scandinavia and its relation to risk intervention campaigns. Nordic Epidemiological SIDS Study. AB - A prospective case-control study of sudden infant death syndrome (SIDS) in Norway, Denmark and Sweden between September 1, 1992 and August 31, 1995 comprised 244 cases and 869 matched controls. After the introduction of risk intervention campaigns, the SIDS incidence decreased from 2.3/1000 live births in Norway, 1.6 in Denmark and 1.0 in Sweden to 0.6/1000 or fewer in all the Scandinavian countries in 1995. The decrease paralleled a decline in the prone sleeping position and there was an accompanying parallel fall in total postneonatal mortality in all three countries. Thus, the risk-reducing campaigns for SIDS have been successful not only in Norway and Denmark, starting from relatively high incidences, but also in Sweden, starting from a low incidence. During the study period, a gradual increase was observed for the effects of prone sleeping, smoking and bottle-feeding as risk factors for SIDS. PMID- 9343278 TI - Quality of life in children with congenital heart defects. AB - Quality of life was measured in children with congenital heart defects (CHDs) registered in a total population of infants born live in the period 1982-91 (n = 22,810), using essential life spheres: external living conditions, interpersonal and personal conditions. In 200 children with CHD alive at the time the investigation was performed, 164 (82%) of the families answered a questionnaire addressing different dimensions of these quality of life spheres. Three subgroups of CHDs were investigated: CHDs spontaneously cured (n = 80), CHDs treated by surgery (n = 56), and CHDs with associated syndromes/malformations (n = 29). 301 (75%) out of 400 controls, matched for age and habitat (county), answered the same questionnaire. The children's ages at investigation were 2 y 2 months-12 y 2 months (median 6 y 1 month). There were no statistically significant differences between the CHD groups and the controls for overall quality of life for any of the three life spheres (p > 0.05). In children with operated CHDs and CHDs associated with syndromes/malformations, quality of life was influenced at some aspects of the external as well as at the interpersonal and personal levels. A trend existed for a higher subjective experience of quality of life in the total CHD group as well as in all the subgroups. It is speculated that this may represent development of coping mechanisms and recalibration of values of life. PMID- 9343277 TI - Behavioral adjustment in children of diabetic mothers. AB - The aim of this study was to evaluate whether maternal diabetes in pregnancy may adversely affect the children's behavioral adjustment, in a sample of 201 mothers (68 with pre-gestational diabetes, 50 with gestational diabetes, and 83 with non diabetic pregnancies) and their singleton offspring. After accounting for socioeconomic status, ethnicity and maternal attitudes, none of the Child Behavior Checklist ratings correlated significantly with maternal patient group or several indices of antepartum maternal metabolism. Child obesity, a common sequela of diabetic pregnancies, correlated positively with Internalizing Behavior problems and three narrow-band sub-scales: Somatic Complaints, Anxious/Depressed, and Social Problems. Results suggest that children of diabetic mothers are at increased risk for a variety of developmental disturbances. Screening for learning and behavioral difficulties should be made at regular pediatric visits, with follow-up evaluations warranted by positive indications, excessive weight gain, or other evolving medical concerns. PMID- 9343279 TI - The burden of nocturnal enuresis. AB - Nocturnal enuresis is a well-known "low-severity high-prevalence" condition in paediatrics, with extensive psychosocial suffering. This suffering is not always realized by paediatricians and other professionals. The aim of this study is to show that enuresis not only has an impact on the child, but also frustrates the entire family. The literature shows that nocturnal enuresis causes distress and low self-esteem for the child. It also has major social and economic implications for the family, with an increasing intolerance as the child grows older. An analysis of nine studies on the impact of successful treatment on the psychological condition of enuretic children showed improved behaviour and personality scores. In five studies the improvement in mental health was significantly related to treatment success. Timely treatment will prevent psychosocial damage, favour a normal development of the child and bring practical relief to the family. PMID- 9343280 TI - Natural odour preferences of newborn infants change over time. AB - At their first sucking contact, neonates prefer an unwashed breast to a washed one, but an amniotic fluid (AF)- treated breast over a "natural odour" breast. We examined the development of these neonatal olfactory preferences. On days 3-4 significantly more babies still selected their mother's unwashed breast (n = 21) than the washed alternative (n = 8). Preferences for natural breast odours were more pronounced for girls than boys. In a subsequent experiment comprising another 28 babies, the number of babies who selected a naturally scented (n = 9) vs an AF-treated breast (n = 19) on days 2-5 were not reliably different. However, babies who selected the natural breast had longer pre-test maternal contact and had spent more time breastfeeding. Ten babies who chose the AF breast in the latter experiment were tested in the same manner several days later; all preferred the naturally smelling breast. While preferences for AF fade after birth, responsiveness to natural breast odours may be enhanced by postnatal experience. PMID- 9343281 TI - Serum levels of magnesium at birth related to complications of immaturity. AB - Magnesium is a natural calcium channel blocker inhibiting vasoconstriction in numerous vascular beds. Magnesium sulphate given prior to birth to pre-eclamptic mothers and mothers in preterm labour has in retrospect been found to be associated with a decreased incidence of both intraventricular haemorrhage and cerebral palsy. Little is known about the effect of normal variations of serum magnesium in the very preterm baby, where morbidity is closely related to rapid vascular changes. We have analysed the absolute levels and normal variations of magnesium concentration in cord blood and during the first 3 weeks after birth for 69 infants born before 32 gestational weeks of age. The results show an inverse relation between serum magnesium at birth and gestational age. Higher levels of serum magnesium at birth within normal variations were associated with a delayed closure of the ductus arteriosus, and mild but not severe peri- and intraventricular haemorrhage. PMID- 9343282 TI - Cerebrospinal fluid plasminogen activator inhibitor-1: a prognostic factor in posthaemorrhagic hydrocephalus. AB - Intraventricular fibrinolytic enhancement with plasminogen activators is an experimental treatment for posthaemorrhagic hydrocephalus, but some infants do not respond. The objectives of this study were to investigate whether plasminogen activator inhibitor-1 is detectable in normal or posthaemorrhagic neonatal cerebrospinal fluid and whether higher neonatal cerebrospinal fluid concentrations of plasminogen activator inhibitor-1 are associated with failure of fibrinolytic therapy. Cerebrospinal fluid samples from 7 controls and 16 infants with posthaemorrhagic hydrocephalus (15 treated with exogenous fibrinolytic agents) were analysed for plasminogen activator inhibitor-1. Plasminogen activator inhibitor-1 was not detectable in any of the control samples but was detectable in all but one of the posthaemorrhagic samples, and at significantly higher levels in the treatment failures (median 94 ng ml(-1)) than in the treatment successes (median 25 ng ml(-1)). High levels of plasminogen activator inhibitor-1 in the cerebrospinal fluid are predictive of, and provide a plausible biological explanation for, failure of intraventricular fibrinolytic therapy. PMID- 9343283 TI - The value of immunoglobulin and complement levels in the early diagnosis of neonatal sepsis. AB - Serum IgG, IgG1, G2, G3, G4, IgM, C3c and C4 concentrations were measured in 24 term neonates with sepsis and 17 healthy normal neonates of similar age, sex and weight (control group). The serum IgG, IgG1, G2, G3, G4, IgM, C3c, and C4 levels were similar in the patients with sepsis and the control group (p > 0.05). In the neonates with sepsis, serum IgG, G1, G2, IgM and C4 levels were not significantly different between the 1st and 10th days, while there were significant differences for IgG3, G4 and C3c (p < 0.05). We conclude that the serum levels of IgG, IgG1, G2, G3, G4, IgM, C3c and C4 concentrations are of no value for the early diagnosis of neonatal sepsis. PMID- 9343284 TI - Hypoantigenic (HA) milk formula and blood cholesterol level in infants at 3 months of age. AB - A preliminary observation is reported concerning total and high-density lipoprotein blood cholesterol levels in 36 infants at 3 months of age fed with a hypoantigenic milk formula. The values are compared with those of 66 breastfed and 76 conventional formula-fed infants. Total and high-density lipoprotein cholesterol levels in hypoantigenic formula-fed infants (152 and 65.7 mg dl(-1), respectively) were comparable to those of breastfed infants (148 and 61.8 mg dl( 1)) and significantly (p < 0.05) higher than those of conventional formula-fed infants (130 and 42.5 mg dl(-1)). PMID- 9343285 TI - Vitamin D status does not influence the breast-milk calcium concentration of lactating mothers accustomed to a low calcium intake. AB - Plasma 25-hydroxy-vitamin D and breast-milk calcium concentration were measured at 3 months of lactation in 60 Gambian mothers accustomed to a low calcium diet, of whom 30 were consuming a calcium supplement and 30 were receiving a placebo, and in 48 British mothers. The plasma 25-hydroxy-vitamin D concentration of the Gambian women was not affected by either calcium supplementation (supplemented, 64.4 +/- 2.5 nmol l(-1); placebo, 64.9 +/- 3.5 nmol l(-1); mean +/- SE) or season. The British average was lower (53.9 +/- 3.0 nmol l(-1), p = 0.004), owing to marked seasonal effects. The breast-milk calcium concentration was lower in The Gambia (supplemented, 5.38 +/- 0.13 mmol l(-1); placebo, 5.10 +/- 0.13 mmol l(-1); British, 6.93 +/- 0.15 mmol l(-1), p < 0.0001). There was no relationship between plasma 25-hydroxy-vitamin D and breast-milk calcium concentration in any group. There was no trend towards lower breast-milk calcium concentration in women with vitamin D status towards the bottom of the normal range or in British women during the winter. This study provides no support for the hypothesis that breast-milk calcium concentration is influenced by vitamin D status or that lactating women with a low calcium intake are at particular risk of vitamin D deficiency. PMID- 9343286 TI - Screening for complement deficiency in bacterial meningitis. AB - Seventy-seven children with bacterial meningitis were screened for complement deficiency. Both the classical and the alternate pathways were normal in 75 patients. Transiently reduced total haemolytic activity of the classical pathway was documented in a boy with meningococcal meningitis. Total haemolytic activity of both the classical and the alternate pathways were reduced in another patient with pneumococcal meningitis: individual complement components determination indicated predominant activation of the alternate pathway. PMID- 9343287 TI - Formal retrospective case review and sudden infant death. AB - A review of 24 consecutive sudden infant deaths was undertaken to evaluate the importance of the various stages in the postmortem assessment of such cases. Death in three cases was caused by obvious trauma. Of the remainder, 16 were attributed to sudden infant death syndrome (SIDS), 4 to accidental asphyxia (identified by death scene examination and/or formal case review) and 1 to a lingual thyroglossal duct cyst. Three (14%) of 21 deaths thought to be SIDS after postmortem examination were attributed to asphyxia following subsequent formal case review. PMID- 9343288 TI - Tyrosinemia type III: diagnosis and ten-year follow-up. AB - Tyrosinemia type III, caused by deficiency of 4-hydroxyphenylpyruvate dioxygenase, is a rare disorder of tyrosine catabolism. Primary 4 hydroxyphenylpyruvate dioxygenase deficiency has been described in only three patients. The biochemical phenotype shows hypertyrosinemia and elevated urinary excretion of 4-hydroxyphenyl derivatives. We report the clinical and biochemical findings and the results of long-term follow-up in a new patient with this disorder presenting with severe mental retardation and neurological abnormalities. The clinical phenotype is compared with those reported in the three previously described patients. PMID- 9343289 TI - Precocious pseudopuberty due to a granulosa cell tumour in a seven-month-old female. AB - We describe a juvenile granulosa cell tumour resulting in pseudopuberty in an infant female. The progression of the clinical signs of puberty were non consonant and the diagnosis was complicated by marginally elevated serum alpha fetoprotein levels. The histological appearance of the resected tumour and binding of MIC2 antibody to tumour cells confirmed the diagnosis. PMID- 9343290 TI - Recurrent ovarian cyst and mutation of the Gs alpha gene in ovarian cyst fluid cells: what is the link with McCune-Albright syndrome? AB - Isolated peripheral precocious puberty due to recurrent ovarian cysts evokes a McCune-Albright syndrome (MAS). This syndrome associates endocrine dysfunction such as precocious puberty, polyostotic fibrous dysplasia, and "cafe-au-lait" skin lesions. We report the case of a 3-y-old girl who presented with peripheral puberty with extremely elevated oestradiol level, low LH and FSH levels, and an ovarian cyst that quickly resolved. Skeletal X-rays were normal and she had no cafe-au-lait spots. GnRH analogue treatment was ineffective. A second ovarian cyst appeared and was completely drained under ultrasonographic guidance. Molecular biological analysis performed on fluid cells revealed the Arg201-->His mutation of the Gs alpha gene described in MAS. Percutaneous aspiration of simple ovarian cyst to detect "MAS" mutation is of interest in the diagnosis of recurrent ovarian cyst. PMID- 9343291 TI - Open letter to scientific journals, the pharmaceutical industry and drugs approval authorities. PMID- 9343292 TI - Plasma thrombomodulin, plasminogen activator and plasminogen activator inhibitor levels in preterm infants with or without respiratory distress syndrome. PMID- 9343293 TI - Is fluid volume depleted in bacterial meningitis? PMID- 9343294 TI - Which hydration in children with acute meningitis? PMID- 9343295 TI - AIDS in Croatia compared with global HIV status. PMID- 9343296 TI - Psychological long-term coping with experience of disease and treatment in childhood cancer survivors. PMID- 9343302 TI - Repression of the acetyl-CoA carboxylase gene in ovine adipose tissue during lactation: the role of insulin responsiveness. AB - We have investigated the mechanisms whereby lipogenesis is markedly suppressed in adipose tissue depots of lactating sheep. Expression of the gene encoding acetyl CoA carboxylase (ACC), the flux-determining enzyme of the lipogenic pathway, is reduced approximately threefold in both omental and subcutaneous adipose tissue depots during late pregnancy and remains so into lactation when compared with non pregnant, non-lactating animals. By comparison, total ACC enzyme activity in these adipose depots is suppressed approximately 25- to 30-fold in lactation. Culture of explants from the subcutaneous depot of lactating sheep with insulin plus dexamethasone for 72 h resulted in an approximately sevenfold increase in ACC mRNA, a fivefold increase in total enzyme activity and a marked increase in the proportion of the enzyme in the active state when compared with explants cultured with no added hormones for the same period. However, there was a lag of between 32 and 48 h before marked induction of any of these parameters by insulin plus dexamethasone was observed. Induction of the alpha-tubulin gene paralleled that of the ACC gene, suggesting that cytoskeletal rearrangements are associated with the aquisition of sensitivity to insulin plus dexamethasone. These results demonstrate that the reduction in lipogenic capacity in ovine adipose tissue during lactation is related to repression of the ACC gene, both at the level of steady-state mRNA abundance and possibly at translation, as well as to suppression of the mechanisms that regulate the proportion of ACC in the active state, and these are further related to the marked insensitivity of these parameters to insulin plus dexamethasone in vitro. PMID- 9343303 TI - Long and short forms of the ovine prolactin receptor: cDNA cloning and genomic analysis reveal that the two forms arise by different alternative splicing mechanisms in ruminants and in rodents. AB - Prolactin is a pituitary hormone that binds to specific receptors in numerous tissues. Depending on the size of their cytoplasmic domain, long and short prolactin receptors (l-PR, s-PR) have been described. Up to now, s-PR were found in rodents only. We report here the cloning of full-length coding sequences for short and long ovine prolactin receptors (s-oPR, l-oPR). The only difference between s- and l-oPR coding sequences was, respectively, the presence or absence of a 39 base pair insert at the beginning of the cytoplasmic domain, with two contiguous inframe stop codons at its 3' end. Sequence comparison revealed that the alternative splicing producing s- and l-oPR was different from that of rodents, although the resulting proteins were very similar. PCR experiments on ovine genomic DNA showed that the 39 base pair insert was directly linked to the downstream exon, and separated from the upstream exon by an 800 base pair intron. Thus, the alternative splicing used a single intron with one 5' and two 3' sites. The same organization was found in bovine and caprine genomes, suggesting that this feature is general in ruminants and different from rodents, which use mutually exclusive exons to produce s-PR and l-PR. PMID- 9343304 TI - Enhancement of ovine trophoblast interferon by granulocyte macrophage-colony stimulating factor: possible involvement of protein kinase C. AB - Interferon-tau (oIFNtau), the major secretory product of ovine conceptuses between days 13 and 21 (day 0=day of estrus) of pregnancy, is implicated in the process of maternal recognition of pregnancy. Culturing of day-14 and day-16 conceptus tissues in the presence of human granulocyte macrophage-colony stimulating factor (hGM-CSF) or interleukin-3 (IL-3) produces a marked increase in oIFNtau mRNA and protein expression. Since GM-CSF and IL-3 are localized at the luminal and glandular epithelia of the ovine endometrium, maternally derived GM-CSF and IL-3 may affect conceptus production of oIFNtau in a paracrine manner. However, the molecular mechanisms by which endometrial GM-CSF and IL-3 up regulate oIFNtau production have not been defined. As an initial investigation of the signaling pathway regulating the GM-CSF induction of the oIFNtau gene, day-16 conceptuses were treated with an inducer, phorbol 12-myristate 13-acetate (PMA) and an inhibitor, calphostin C of the protein kinase C (PKC) pathway. Treatment with either 150 units/ml hGM-CSF (P<0.01) or 10 nM PMA (P<0.05) resulted in a significant increase in oIFNtau mRNA expression. Pretreatment of conceptuses with 1 microM PMA for 12 h to produce PKC-deficient tissues or treatment with 50 mM calphostin C abolished the hGM-CSF-induced increase in oIFNtau mRNA. An in vitro expression system was established for the analysis of oIFNtau gene regulatory sequences. The oIFNtau010 gene has been isolated previously and found to be the principal oIFNtau gene up-regulated during the preimplantation period. 5' Flanking regions of the oIFNtau010 gene, 2 kb and 0.8 kb, were cloned into a basic chloramphenicol acetyltransferase reporter plasmid. These oIFNtau010 promoter constructs, along with expression controls, were transfected into human choriocarcinoma cells (JAR and JEG3) and their responsiveness to hGM-CSF and second messenger system activators including PMA, calcium ionophore (A23187) and 8-bromo-cAMP were characterized. The oIFNtau010 promoter constructs were up regulated by hGM-CSF and PMA treatments (P<0.01). Combined treatment with PMA and A23187 prevented the promoter activation seen with PMA alone. The conceptus culture data, along with the results from the transfection experiments, suggest that the stimulatory effect of GM-CSF on oIFNtau is mediated through the PKC second messenger system. PMID- 9343305 TI - The regulation of mammary prolactin receptor metabolism by a retroviral envelope protein. AB - In a previous study, infection with the mouse mammary tumor virus (MMTV) was shown to increase the sensitivity of the mammary epithelium toward prolactin (PRL); furthermore, this effect could be mimicked by the binding of the MMTV envelope protein (gp52) to its cell receptor. The present work has investigated the possibility that gp52-induced changes in the PRL receptor (PRLR) were responsible for this phenomenon. In vitro, gp52 doubled the PRLR concentration in the plasmalemma of mammary epithelium without affecting the affinity. The origins of these PRLRs were twofold: first, gp52 stimulated PRLR mRNA nearly fivefold, suggesting that some of the receptors were newly synthesized. Second, there was a redistribution of PRLRs within the mammary cell: PRLRs were shifted from an internal pool to the plasma membrane. This relocation was very rapid, occurring within 30 min. There did not appear to be any contribution from alterations in PRLR degradation, since the half-life of PRLR was not affected by gp52. In summary, the MMTV increases the PRL sensitivity of mouse mammary epithelium by elevating PRLRs through both enhanced synthesis and recruitment from microsomes. PMID- 9343306 TI - 3,5-Diiodo-L-thyronine (T2) has selective thyromimetic effects in vivo and in vitro. AB - Recent data have suggested that the iodothyronine, 3,5-diiodo-l-thyronine (T2), has selective thyromimetic activity. In vivo, T2 has been shown to suppress TSH levels at doses that do not produce significant peripheral manifestations of thyroid hormone activity. Furthermore, T2 has been shown to produce smaller increments in peripheral indices of thyroid status than does T3, when doses resulting in equivalent suppression of circulating TSH are compared. We have assessed the selective thyromimetic activity of T2 in vivo and in vitro, and performed in vitro studies to assess the potential molecular basis for these selective properties. T2 was 100-fold less potent than T3 in stimulating GH mRNA levels in GH3 cells. In contrast, the iodothyronines were almost equivalent in their ability to downregulate TRbeta2 mRNA levels in this cell line. Both 3,3' diiodo-L-thyronine and thyronine exhibited no significant thyromimetic effects on either process. In vivo, doses of T2 and T3 that were equivalent in their induction of hepatic malic enzyme (ME) mRNA did not produce equivalent suppression of circulating TSH, with T2 being only 27% as effective as T3. T2 was up to 500-fold less potent than T3 in displacing [125I]-T3 from in vitro translated specific nuclear receptors (TRs) and GH3 cell nuclear extracts. Electrophoretic mobility shift assays, assessing the ability of T2 to produce dissociation of TRbeta1 homodimers from inverted palindrome T3 response elements, indicated that T2 was also 1000-fold less potent than T3 in this respect. These data confirm that T2 has significant thyromimetic activity, and that this activity is selective both in vivo and in vitro. However, there are no data to support a selective central effect, T2 being relatively more potent in stimulating hepatic ME mRNA than in suppression of TSH in vivo. The basis for this differential thyromimetic activity is not selective affinity of the different TR isoforms for T2, or divergent properties of T2 in competitive binding and functional assays in vitro. PMID- 9343307 TI - Distinct promoter sequences of two precursor genes for salmon gonadotropin releasing hormone in masu salmon. AB - Two types of genes encode salmon gonadotropin-releasing hormone (sGnRH), which is thought to act on both sexual maturation and reproductive behavior, in salmonids. We characterized the two sGnRH genes (sGnRH-I and -II) and their upstream regions in masu salmon, Oncorhynchus masou, since such information is a prerequisite for molecular approaches to salmon reproduction. The two genes have similar exon intron structures composed of four exons and three introns. Sequence analyses of the two genes showed that coding regions are highly conserved, but upstream regions are distinctively divergent. In the upstream regions, only the sGnRH-II gene has a large palindromic sequence, which has been proposed to be involved in control of transcription via estrogen receptors. In contrast, the sGnRH-I gene is missing the large palindromic sequence, but has three distinct palindromes in the upstream region. These results may suggest divergent transcription regulatory mechanisms between the two sGnRH genes in masu salmon. The differences in the upstream regions of sGnRH genes in Atlantic salmon (Salmo salar), sockeye salmon (Oncorhynchus nerka) and masu salmon are discussed with respect to the evolution of sGnRH genes in salmonid fish. PMID- 9343308 TI - The nuclear orphan receptors COUP-TFII and Ear-2 act as silencers of the human oxytocin gene promoter. AB - We have previously shown that COUP-TFII and Ear-2, two members of the nuclear orphan receptor family, are able to repress oestrogen-stimulated transcriptional activity of the human oxytocin (OT) gene promoter by binding to a site that overlaps with the oestrogen response element (ERE) present in the 5' flanking region of the gene. Although most nuclear receptor-mediated transcriptional repression conforms with the paradigm of passive repression and involves competitive binding to an activator site, active repression, i.e. silencing of basal promoter activity, has been observed in a limited number of cases. Here we show by co-transfection experiments using COUP-TFII and Ear-2 expression vectors and reporter constructs containing OT gene promoter fragments linked to the chloramphenicol acetyltransferase gene that both COUP-TFII and Ear-2 are capable of silencing basal OT gene promoter activity by 54 and 75% respectively. 5' Deletion and footprint analyses revealed two areas of functionally important interaction sites: (1) a direct TGACC(T/C) repeat overlapping the ERE and (2) a more promoter-proximal area centred at - 90 containing three imperfect direct repeats (R1-R3) spaced by four nucleotides each. Mutagenesis of reporter constructs as well as electrophoretic mobility-shift assays demonstrated that each of the three proximal repeats R1-R3 contributed to orphan receptor binding and the silencing effect. Inasmuch as the orphan receptor-binding sites are not involved in mediating basal transcriptional activity of the OT gene promoter, the observed effects are best interpreted as active repression or promoter silencing. Moreover, since COUP-TFII and Ear-2 are both co-expressed in OT-expressing uterine epithelial cells, the novel transcriptional effects described here are likely to be of functional importance in the fine-tuning of uterine OT gene expression in vivo. PMID- 9343309 TI - Expression of neurotrophin-3 in the growing velvet antler of the red deer Cervus elaphus. AB - Antlers are organs of bone which regenerate each year from the heads of male deer. In addition to bone, support tissues such as nerves also regenerate. Nerves must grow at up to 1 cm/day. The control of this rapid growth of nerves is unknown. We examined the relative expression of neurotrophin-3 (NT-3) mRNA in the different tissues of the growing antler tip and along the epidermal/dermal layer of the antler shaft of the red deer Cervus elaphus, using semi-quantitative reverse transcription-polymerase chain reaction. Expression in the tip was found to be highest in the epidermal/dermal layer and lowest in the cartilaginous layer in all developmental stages examined. These data correlate well with the density and pattern of innervation of these tissues. Along the epidermal/dermal layer of the antler shaft, expression was highest in the segments subjacent to the tip and lowest near the base, arguing for differences in the temporal expression of NT-3 in these segments. The expression of NT-3 in cells isolated from the different layers of 60-day antlers did not mirror that observed when whole tissues were used and may suggest regional specificity of NT-3 expression within antler tissues. PMID- 9343310 TI - Recognition of follicle stimulating hormone (alpha-subunit) by a recombinant receptor protein domain coded by an alternately spliced mRNA and expressed in Escherichia coli. AB - To assess the functional significance of putative proteins encoded by alternately spliced mRNA of the sheep testicular FSH receptor, a short form cDNA comprising of the first four exons (117 residues mature protein) was engineered for expression in Escherichia coli. The expressed protein of molecular mass 15 kDa was purified to homogeneity and verified by reaction with an antibody against a synthetic peptide sequence unique to the amino (N)-terminal region FSH receptor. The purified FSH receptor domain protein bound 125I-labeled hFSH in a ligand blot on polyvinylidine difluoride membranes. Further analyses by slot blot revealed high affinity of the immobilized protein with significant reaction at 10 pmol. As the immobilized receptor protein also reacted with structurally related hormones (125I-labeled LH/125I-labeled human chorionic gonadotropin), we confirmed that interaction most probably occurred via the common alpha-subunit of these glycoprotein hormones. Our results reveal that this N-terminal portion of the FSH receptor contain(s) major site(s) for hormone recognition that could be mediated via the alpha-subunit. A rabbit antibody to the receptor inhibited FSH action in receptor bearing cells, revealing the utility of such recombinant FSH receptor protein(s) for modulation of hormone action. PMID- 9343311 TI - Expression of membrane and soluble intercellular adhesion molecule-1 in Graves' disease. AB - We have investigated the in vitro expression of membrane and soluble intercellular adhesion molecule-1 (ICAM-1) by human thyroid cells from 20 patients with Graves' disease and 5 normal subjects. Membrane ICAM-1 was not detected by flow cytometry analysis in non-cultured thyrocytes from either normal or Graves' disease tissues. It appeared on thyroid cells after a 24-h culture in monolayers and showed a regular dose-dependent increase. The same results were obtained with soluble ICAM-1 (sICAM-1) in culture media from cells cultured in monolayers, vesicles or follicles. No change was obtained with different concentrations of fetal calf serum added to the media. Coculture of Graves' disease thyrocytes with autologous peripheral blood lymphocytes (PBL) or intrathyroidal lymphocytes (ITL) enhanced the expression of both membrane and sICAM-1 whatever the culture model. When normal thyrocytes were cocultured with PBL, sICAM-1 increased but with ITL sICAM-1 remained unchanged. High concentrations of gamma interferon induced an increase of both membrane and sICAM 1 in the three culture models. However the increases were greater with vesicles and follicles. Only sICAM-1 levels were raised with 0.1, 1 and 10 microM retinoic acid. These results suggest that ICAM-1 appears in culture, possibly due to mechanical effects such as adherence to plates and cell-to-cell contacts. Moreover, its expression is modulated by several factors such as cytokines or retinoic acid. Further investigations are needed to establish whether ICAM-1 is really involved in the pathogenesis of Graves' disease. PMID- 9343312 TI - Alternate splicing of the RNA for hamster type 2 3 beta-hydroxysteroid dehydrogenase/delta 5-->4isomerase. AB - Complementary DNAs encoding the hamster type 2 3 beta-hydroxysteroid dehydrogenase/delta 5-->4 isomerase were isolated from liver and kidney cDNA libraries. Nine clones were isolated containing identical coding and 3' untranslated sequences. However, six of the clones contained a 68-nucleotide stretch in the 5' untranslated region that was missing in the other three clones. Primers were designed to flank this region and the polymerase chain reaction (PCR) was performed on hamster liver and adrenal RNA. Two PCR products were amplified of the predicted molecular sizes and with the expected sequence. Primers were then designed to amplify sequences encompassing this region from hamster genomic DNA. Sequencing of the resultant PCR products demonstrated that the 68-nucleotide stretch missing in some transcripts corresponded exactly to the second of three exons identified. We conclude that the 5' untranslated region of this mRNA is transcribed from at least three exons, and that the sequence of the second of these exons is spliced out of some of the RNA transcripts. PMID- 9343313 TI - Use of a newly developed rapid microbial ATP bioluminescence assay to detect microbial contamination on poultry carcasses. AB - A newly developed rapid microbial ATP bioluminescence test (R-mATP) was shown to be an adequate means to assay the microbial load of poultry carcasses. This assay utilizes differential extraction and filtration to separate somatic from microbial ATP in a very rapid timeframe. The assay requires approximately 5 min to complete; approximately 3.5 min to sample and 90 s analytical time. Correlation coefficient (r) between aerobic colony counts and R-mATP test results (n=329) was 0.82. Post-test probabilities to correctly classify carcasses with different levels of microbial contamination were as high as 98% for samples of > or = 3.5 log aerobic CFU per ml. Given the rapidity of this assay, the R-mATP holds potential for monitoring the microbial load of carcasses at poultry processing critical control points. Other potential applications of this new version of the microbial ATP bioluminescence test are discussed. PMID- 9343314 TI - Effect of polymers on enhanced chemiluminescent assays for peroxidase and peroxidase labels. AB - Hydroxypropyl methylcellulose, hydroxyethyl cellulose, and hydroxybutyl methylcellulose stabilized light emission in a boronic acid-enhanced chemiluminescent assay for horseradish peroxidase. The stabilization of light emission was concentration-dependent and more effective with substituted boronic acid enhancers (e.g. 4-iodophenylboronic acid) than with substituted phenol enhancers (e.g. 4-iodophenol). Hydroxybutyl methylcellulose improved the linearity of the dose-response curve in a peroxidase-based antioxidant assay and stabilized light emission post-consumption of the antioxidant (Trolox). This polymer had no effect on the signal from a peroxidase label immobilized on a membrane (dot blot) or on the inside surface of a microwell in an enzyme immunoassay for thyrotropin. PMID- 9343315 TI - Amines for detection of dopamine by generation of hydrogen peroxide and peroxyoxalate chemiluminescence. AB - A range of nitrogen-containing compounds (alkyl amines, piperazines, cyclohexylamines and nitrogen heterocyclics) were investigated for generation of hydrogen peroxide from dopamine and detection by peroxyoxalate chemiluminescence. Imidazole, ethyleneurea and allantoin among the nitrogen heterocyclic compounds tested generated hydrogen peroxide from dopamine following incubation at 60 degrees C, pH 9.5-10.5, for 0-30 min. Imidazole was the most effective for generation of hydrogen peroxide, but imidazole derivatives with a primary amine side chain (histamine) or thiol (ethylenethiourea) were not effective. The presence of a ketone group (ethyleneurea, allantoin) did not hinder the reaction. Under optimal conditions (30 min incubation, 50 mmol/L imidazole) 10.5 nmol of dopamine could be detected. The cyclohexylamines tested produced low amounts of hydrogen peroxide (0.09-2.74% of light intensity with imidazole), and the piperazines and the alkyl amines tested produced no detectable hydrogen peroxide. Imidazole reacts with the phenolic groups of dopamine in a different manner from monoamine oxidase, and a reagent containing imidazole, ethyleneurea or allantoin was useful for non-enzymatic detection of dopamine by peroxyoxalate chemiluminescence. PMID- 9343316 TI - Industry--a vital partner for academic medicine. PMID- 9343317 TI - Recommendations for the reporting of tissues removed as part of the surgical treatment of cutaneous melanoma. Association of Directors of Anatomic and Surgical Pathology. AB - The Association of Directors of Anatomic and Surgical Pathology have developed recommendations for the surgical pathology report for common malignant tumors. The recommendations for cutaneous melanoma are reported herein. PMID- 9343318 TI - Parietal cell protrusions in gastric ulcer disease. AB - Oxyntic mucosal biopsy specimens from patients receiving omeprazole therapy have been described as frequently showing characteristic tonguelike protrusions of parietal cell cytoplasm (PCP) into the gland lumen. Although protrusion of parietal cell cytoplasm is believed to be associated with omeprazole therapy and has been implicated in the histogenesis of fundic gland polyps, we have observed it in a wide variety of different conditions unrelated to peptic ulcer disease or omeprazole therapy. To establish the incidence of PCP and analyze its relationship to gastritis, gland dilatation, cystic change, and fundic gland polyps, we studied 400 gastric mucosal biopsy specimens from gastric ulcer patients who were not receiving omeprazole therapy and who did not receive any medications for at least 2 weeks. Severity of each of these changes was graded on a scale of I to III. PCP was observed in oxyntic mucosal biopsy specimens from 60 (15%) patients and was associated with varying grades of chronic superficial or interstitial gastritis in 25 (Helicobacter pylori was identified in 12). Although chronic atrophic gastritis, cystic change, or fundic gland polyps were not identified in any of the cases with PCP, gland dilatation was present in 25 of 60 (42%) biopsy specimens. No consistent linear correlation was observed between increasing grades of PCP and gastritis or gland dilatation. Our findings of PCP in 15% of gastric ulcer patients who were off all medications for 2 weeks indicate that PCP is not always related to omeprazole usage. It appears to be a change encountered in a wide variety of diverse settings and, therefore, should not be used to monitor omeprazole therapy. In gastric ulcer patients, there is no linear correlation between PCP and gland dilatation or severity of gastritis. The lack of association of PCP with such cardinal features of fundic gland polyps as gland dilatation and cystic change suggests that PCP per se has little if any role in the development of such polyps. The exact clinical and functional significance of PCP remain to be established and merits further investigation. PMID- 9343319 TI - Coordinated expression of integrin alpha6beta1 and laminin in hepatocellular carcinoma. AB - The interaction between tumor cells and laminin mediated by laminin-binding integrins is critical for tumor invasion and metastasis. The aim of this study was to clarify the altered expression of laminin-binding integrins with the change of laminin deposition in hepatocellular carcinoma (HCC) in comparison with cirrhotic or normal liver by immunohistochemistry. In HCC, hepatoma cells and sinusoidal endothelial cells expressed integrins alpha1beta1, alpha2beta1, alpha3beta1, and alpha6beta1. Integrins alpha1beta1 and alpha6beta1 were detected in a continuous pattern along the sinusoids in accordance with laminin assembly. Integrins alpha2beta1 and alpha3beta1 were detected in a discontinuous pattern at these sites. Integrin alpha6beta4 was not detected. In cirrhotic liver, although integrins alpha1beta1 and alpha6beta1 as well as laminin were detected in a continuous pattern along the sinusoids, integrins alpha2beta1, alpha3beta1, and alpha6beta4 were not detected. In normal liver, although integrin alpha1beta1 was detected in a continuous pattern along the sinusoids, neither integrins alpha2beta1, alpha3beta1, alpha6beta1, alpha6beta4, nor laminin were detected. We have clarified that, of laminin-binding integrins, the localization of integrin alpha6beta1 shows the best correspondence with the localization of laminin. These results suggest that of laminin-binding integrins, integrin alpha6beta1 is very important for cell-laminin interactions in HCC. PMID- 9343320 TI - Computerized microscope morphometry of umbilical vessels from pregnancies with intrauterine growth retardation and abnormal umbilical artery Doppler. AB - Computerized microscope morphometry was used to study cross sections from the vessels of the umbilical cord in placentas of patients with intrauterine growth retardation (IUGR) that displayed either normal or abnormal umbilical arteries (UA) Doppler flow velocity waveforms (FVW). Cords from 63 eutrophic fetuses with normal Doppler (controls), 47 IUGR fetuses with normal Doppler and 32 IUGR fetuses with abnormal Doppler underwent morphometric analysis using a highly optimized microscope environment (HOME) and "CordHOME" software. IUGR with an accompanying normal Doppler versus control showed a reduction of Wharton jelly and both the total and lumen vein areas. IUGR with an accompanying pathological Doppler showed a comparable reduction in wall thickness and areas of every vessel. These findings indicate that the hypoplastic umbilical vessels are associated with an increase in placental vascular resistance that may be the consequence of underdevelopment in response to a chronic reduction in placental blood flow. PMID- 9343321 TI - Platelet-derived endothelial cell growth factor/thymidine phosphorylase immunohistochemical expression in lymphoid tissue and lymphoid malignancies. AB - The catabolic enzyme thymidine phosphorylase (TP) plays a crucial role in nucleic acid metabolism by regulating the availability of thymidine. Platelet-derived endothelial cell growth factor (PD-ECGF) is an angiogenic factor that was recently shown to be TP. The angiogenic properties of PD-ECGF/TP are attributable to a reduction of thymidine levels that results in a promotion of endothelial cell proliferation. Early studies showed a higher concentration of TP in macrophages than in parenchymal cells and in neoplastic than in nonneoplastic tissues. We examined the immunohistochemical expression of PD-ECGF/TP in reactive lymphoid tissues (lymph node and tonsil), as well as in a series of 20 cases of Hodgkin's disease and 31 cases of non-Hodgkin's lymphomas. Macrophages, sinus lining cells, and cells with dendritic morphology, of both follicular dendritic and interdigitating reticular cell type, presented a prominent nuclear and cytoplasmic positivity in reactive lymphoid tissue and in malignant lymphomas. Small lymphocytes and the neoplastic population were always negative, whereas endothelial staining was variable and showed no correlation to the type or grade of the lymphomas. In Hodgkin's disease (with the exception of the nodular lymphocyte predominance type) and some cases of high-grade non-Hodgkin's lymphomas, the positive dendritic cells formed a dense meshwork closely surrounding the neoplastic population. Our results suggest that the reported upregulation of PD-ECGF/TP activity in lymphoid malignancies is attributable to the nonneoplastic population, especially to cells of dendritic morphology. PMID- 9343322 TI - Differentiation of colonic metaplasia from adenocarcinoma of urinary bladder. AB - Colonic metaplasia and primary bladder adenocarcinoma are relatively uncommon entities that can have similar gross clinical appearances. Examples of colonic metaplasia histologically mimicking cancer have only rarely been reported. We retrospectively analyzed 38 cases of cystitis glandularis (18 cases of colonic metaplasia), 12 cases of adenocarcinoma of urinary bladder (two well differentiated, WDA), and one in situ adenocarcinoma from the surgical pathology files of Johns Hopkins Hospital. Nine patients with colonic metaplasia had widespread lesions. Two showed superficial muscularis propria involvement, mimicking adenocarcinoma; one of these cases had been diagnosed as infiltrating WDA at both an academic center and a community hospital. Dissecting mucin pools were focally seen in four cases of widespread colonic metaplasia, also mimicking cancer. One of the nine cases showed minimal cytological atypia, but no cases showed mitoses or signet ring cells. Distinguishing WDA from colonic metaplasia was the finding in WDA of infiltrative architectural pattern (two of two), extensive muscle invasion (two of two), moderate anaplasia (one of two), mitotic figures (two of two), and extensive mucinous pools (one of two). The diagnosis of adenocarcinoma in situ was based on anaplasia. Clinically, colonic metaplasia may resemble cancer. Histologically, colonic metaplasia may mimic cancer based on extensive involvement of the lamina propria, focal mucinous pools, focal muscularis propria involvement, focal mild cytological atypia, and rare mitoses. Despite overlapping features with colonic metaplasia, the diagnosis of WDA is based on the greater degree and extent of these atypical findings in cancer. PMID- 9343324 TI - Clonal analysis of gliomas. AB - In malignant gliomas, the characteristically heterogeneous features and frequent diffuse spread within the brain have raised the question of whether malignant gliomas arise monoclonally from a single precursor cell or polyclonally from multiple transformed cells forming confluent clones. Although monoclonality has been shown in surgically resected tissues, these may not include the full spectrum of patterns seen on autopsy material. Little is known about the clonality of low-grade gliomas from which malignant gliomas may sometimes arise. We sought to investigate the clonality of low-grade and malignant gliomas by using and comparing surgical and autopsy material with a Polymerase chain reaction (PCR)-based assay for nonrandom X chromosome inactivation. For that, purpose, archival surgical and autopsy material from 15 female patients (group A) (age 4 to 73 years; median, 45) with malignant gliomas (12 glioblastomas, one gliosarcoma, one anaplastic oligoastrocytoma, one gliomatosis cerebri), surgical material only from 21 female patients (group S) (age 6 to 78 years; median, 60) with low-grade and malignant gliomas (four low-grade astrocytomas, three oligoastrocytomas, two anaplastic astrocytomas, one gemistocytic astrocytoma, four oligodendrogliomas, seven glioblastomas) were analyzed. In group A, representative areas (mean = 5/patient; median = 7) were microdissected from tissue sections and assayed by PCR amplification of a highly polymorphic microsatellite marker locus of the human androgen receptor gene (HUMARA) in the presence of alpha32P with and without predigestion with a methylation-sensitive restriction enzyme (HhaI). Products were resolved by denaturing gel electrophoresis and autoradiographed. In group S, selected tumor areas were used for the assay. Each patient's normal brain tissue was used for control. The band intensity of alleles were measured by densitometric scanning. In group A, 13 of 15 cases were informative (heterozygous). The same pattern of nonrandom X chromosome inactivation was present in all areas of solid dense and moderate tumor infiltration in eight including all components of the gliosarcoma. Two of eight also showed focal loss of heterozygosity (LOH). One of 13 presented global LOH. Two of 13 showed microsatellite instability, one of which in a patient with Turcot syndrome, the other in gliomatosis cerebri. Opposite skewing patterns were seen in distant areas of gliomatosis cerebri consistent with oligoclonal derivation. Clonality remained indeterminate in one glioblastoma and in the anaplastic oligoastrocytoma because of skewed lyonization in the normal control. In group S, 19 of 21 cases were informative. Fifteen of 19 were monoclonal (four low-grade astrocytomas, one anaplastic astrocytoma, one gemistocytic astrocytoma, two oligodendrogliomas, one oligoastrocytoma, six glioblastomas). Four of 19 were indeterminate. We conclude that (1) Low-grade and malignant gliomas are usually monoclonal tumors, and extensively infiltrating tumors must result from migration of tumor cells (2) Gliomatosis cerebri may initiate as an oligoclonal process or result from collision gliomas (3) Biphasic gliomas likely arise from a single precursor cell. (4) LOH at the HUMARA locus is probably related to partial or complete deletion of an X-chromosome, which occurs in malignant gliomas during clonal evolution. PMID- 9343323 TI - MIC2, TdT, bcl-2, and CD34 expression in paraffin-embedded high-grade lymphoma/acute lymphoblastic leukemia distinguishes between distinct clinicopathologic entities. AB - We propose that 12E7 (CD99) expression, along with TdT, bcl-2, and CD34 reactivity in lymphoblastic lymphoma (LyL)/acute lymphoblastic leukemia (ALL), distinguishes this group of neoplasms from small noncleaved cell lymphomas (SNCLs) in both pediatric and adult patients, thereby narrowing the differential diagnosis of high-grade non-Hodgkin's lymphomas and acute lymphoblastic leukemias in paraffin sections. 12E7 (CD99) is one of a group of available antibodies that recognizes the product of the mic-2 gene, which was originally identified in ALL. Despite this, most clinicopathological research has focused on the reactivity of 12E7 in a subset of the small round cell tumors of childhood. Although TdT is widely used in the subtyping of blastic leukemias, its use in the distinction of high-grade lymphomas in paraffin sections has been limited. We collected 24 cases of LyL/ALL (13 B-cell and 11 T-cell) and 15 cases of SNCL from 1984 through 1993. We confirmed the diagnoses using morphology and analysis of immunologic data. We performed immunohistochemistry with the 12E7 antibody, TdT, bcl-2, and CD34 on formalin-fixed, paraffin-embedded material. The patients' ages ranged from 4 to 81 years; nine of the study patients were children. Sixteen of the 24 LyL/ALLs stained with 12E7. In contrast, none of the 15 cases of SNCL reacted with this antibody (chi-square P < .0001). A larger percentage of T-cell LyL/ALLs reacted with 12E7 than did B-cell LyL/ALLs (82% v 54%). Sixteen of 20 LyL/ALLs reacted with the anti-TdT antibody, as compared with none of 11 SNCLs (chi-square P < .0001). Six LyL/ALLs were CD34 positive (of 23), and none of the SNCLs were CD34 positive (0 of 12) (chi-square P = .0519). Bcl-2-positive cases were found among both LyL/ ALLs and SNCLs, although they were more prevalent among LyL/ ALLs (92% v 25%; chi-square P < .0001). When one considers the differential diagnosis of a high-grade lymphoma/acute lymphoblastic leukemia, positive reactions with 12E7, TdT, bcl-2, and CD34 support the diagnosis of LyL/ALL over SNCL. Moreover, we present data that suggests that evaluating for TdT in formalin-fixed paraffin embedded tissue is a more sensitive test than using either 12E7, bcl-2 or CD34 alone. PMID- 9343325 TI - Human DNA topoisomerase II-alpha: a new marker of cell proliferation in invasive breast cancer. AB - DNA topoisomerase II-alpha is the molecular target of doxorubicin, an active drug used in the therapy of breast cancer. From many in vitro studies, it is known that high levels of topo II-alpha expression correlate with drug sensitivity, and low levels of topo II-alpha correlate with drug resistance. In addition, the enzyme is known to be a marker of cell proliferation in normal tissues. Because the number of proliferating cells in a breast cancer has been shown to be prognostically important, and because doxorubicin is used in the treatment of breast cancer, we hypothesized that the measurement of topo II-alpha in breast cancer may not only give drug sensitivity information but also may yield important data on cell proliferation. In this study, formalin-fixed, paraffin embedded tissue from 30 specimens of invasive breast cancer from 20 patients were immunohistochemically stained for topo II-alpha with a mouse monoclonal antibody. For each case, a topo II-alpha index was determined that represents the number of positive-staining tumor cells divided by the total number of tumor cells counted times 100. A similar index was determined for MIB1, a known cell proliferation marker. Each case was also graded according to the modified Bloom-Richardson criteria and evaluated for c-erbB-2 amplification, hormonal status, S-phase fraction, and mitotic index. The topo II-alpha index correlates better with the MIB1 index than with the S-phase fraction or mitotic index. The topo II-alpha expression in breast cancer ranges from low (topo II-alpha index <1) to high (topo II-alpha index = 86). Amplification of c-erbB-2 was observed in 4 of 28 cases (14%) but did not correlate with high topo II-alpha indices. We conclude that measurement of topo II-alpha in invasive breast cancer can be readily performed by immunohistochemical staining, and it gives information on the number of cycling tumor cells. In addition, because the enzyme is the molecular target of doxorubicin, the expression of the enzyme may relate also to the sensitivity or resistance of the tumor to doxorubicin-based chemotherapeutic protocols. PMID- 9343326 TI - The role of p53, p21WAF1/C1PI, and bcl-2 in radioresistant colorectal carcinoma. AB - Genetic alterations in the p53 tumor suppressor gene are common in human colorectal cancers, occurring in approximately 70% of tumors. In vitro studies have shown that wild-type p53 is involved in controlling cell cycle checkpoint functions and apoptosis involved in the cytotoxic response induced by ionizing radiation and several anticancer chemotherapeutic agents. Wild-type p53 protein can transcriptionally activate the WAF gene, which encodes a cyclin-dependent kinase inhibitory protein, p21WAF1/C1PI protein, and transcriptionally repress the bcl-2 gene, which encodes an inhibitor of apoptosis. To learn more about the in vivo relationship between p53 protein and the expression of p21WAF1/C1PI and bcl-2 proteins in human colorectal cancers treated with radiation therapy, we examined the expression of these proteins by immunohistochemistry in pre irradiated biopsy specimens and surgical specimens with residual tumor of 27 patients with colorectal carcinoma. Cell proliferation was measured using Ki-67 expression in the tumor cells. The p53 protein was not detected in normal colorectal mucosa, but it was expressed in 21 of 27 (78%) of pre-irradiated tumor samples and in 19 of 27 (70%) of post-irradiated tumors. Expression of the bcl-2 protein in normal colorectal mucosa was confined to the basal epithelial cells of the crypts. Diffuse bcl-2 staining was detected in tumor cells in 13 of 27 (48%) of pre-irradiated samples and in 14 of 27 (52%) of post-irradiated samples. p21WAF1/C1PI expression was detected in 14 of 27 (52%) of pre-irradiated samples but only in 7 of 27 (26%) of post-irradiated samples. No inverse relationship between expression of p53 protein and abnormal bcl-2 expression was apparent. p21WAF1/C1PI was expressed in most nonproliferating Ki-67-negative epithelial cells at the apical tips of the crypts in normal colorectal mucosa, but not in proliferating Ki-67-positive cells of adjacent adenomatous mucosa. An inverse relationship between Ki-67 and p21WAF1/C1PI expression was observed in normal colorectal mucosa and adjacent adenomatous mucosa. After radiation therapy, p53 protein accumulation did not change among residual tumors in 18 cases (three of which were initially negative and remained negative); in four cases there was a significant increase, and five cases had a substantial decrease of p53 expression. Aberrant bcl-2 expression is not correlated with expression of p53 and does not increase significantly in post-irradiated tumor cells. p21WAF1/C1PI expression is markedly reduced in tumor cells that survive radiation therapy. PMID- 9343327 TI - Significant reduction in the rate of false-negative cervical smears with neural network-based technology (PAPNET Testing System). AB - False-negative cervical Pap smears may lead to disability or death from carcinoma of the uterine cervix. New computer technology has led to the development of an interactive, neural network-based vision instrument to increase the accuracy of cervical smear screening. The instrument belongs to a new class of medical devices designed to provide computer-aided diagnosis (CADx). To test the instrument's performance, 487 archival negative smears (index smears) from 228 women with biopsy-documented high-grade precancerous lesions or invasive cervical carcinoma (index women) were retrieved from the files of 10 participating laboratories that were using federally mandated quality assurance procedures. Samples of sequential negative smears (total 9,666) were retrieved as controls. The instrument was used to identify evidence of missed cytological abnormalities, including atypical squamous or glandular cells of undetermined significance (ASCUS, AGUS), low-grade or high-grade squamous intraepithelial lesions (LSIL, HSIL) and carcinoma. Using the instrument, 98 false-negative index smears were identified in 72 of the 228 index women (31.6%, 95% confidence interval [CI]: 25% to 38%). Disregarding the debatable categories of ASCUS or AGUS, there were 44 women whose false-negative smears disclosed squamous intraepithelial lesions (SIL) or carcinoma (19.3%; 95% CI: 14.2% to 24.4%). Unexpectedly, SILs were also identified in 127 of 9,666 control negative smears (1.3%; 95% CI: 1.1% to 1.5%). Compared with historical performance data from several participating laboratories, the instrument increased the detection rate of SILs in control smears by 25% and increased the yield of quality control rescreening 5.1 times (P < 0.0001). These data provide evidence that conventional screening and quality control rescreening of cervical smears fail to identify a substantial number of abnormalities. A significant improvement in performance of screening of cervical smears could be achieved with the use of the instrument described in this report. PMID- 9343328 TI - Solitary fibrous tumor of urinary bladder: report of two cases. AB - The clinical and pathologic features of two cases of solitary fibrous tumor arising from urinary bladder wall are described. To our knowledge, solitary fibrous tumors have not been previously reported at this site. Both tumors showed typical histologic features of solitary fibrous tumor, were CD34 immunostain positive and pursued a benign clinical course on short term follow-up. PMID- 9343329 TI - Ovarian Sertoli-Leydig cell tumor: a SRY gene-independent pathway of pseudomale gonadal differentiation. AB - Sertoli-Leydig cell tumors (SLCT) are rare sex-cord stromal tumors of the ovary composed of undifferentiated gonadal stromal cells, Leydig cells (LC), and Sertoli cells (SC), with the latter forming structures resembling fetal testicular tubules. The histogenetic basis of morphological male differentiation patterns in females is controversial. Here, we report a SLCT with intermediate differentiation in a 23-year-old woman investigated by light microscopy, immunohistochemistry for intermediate filaments, and sex steroid hormone receptors (SSHR), as well as by polymerase chain reaction (PCR) for the presence of the sex-determining region Y gene (SRY). Our investigation shows that the SCs of SLCT express progesterone and androgen receptors as well as cytokeratins and vimentin. By PCR, SLCT-derived genomic DNA lacked the SRY gene, indicating that the SLCT results from a SRY gene-independent pathway of pseudomale gonadal differentiation. The expression of progesterone receptors (PRs) in the SCs of the SLCT is in contrast to their absence in testicular SCs, but in line with their presence in ovarian granulosa and surface epithelial cells. Thus, our results provide strong evidence for a close histogenetic relationship between the SLCT and the female gonocyte-supporting cell, the granulosa cell (GC). PMID- 9343330 TI - Thymoma associated with CD4+ lymphopenia, cytomegalovirus infection, and Kaposi's sarcoma. AB - A case of thymoma with associated opportunistic infections, CD4/CD8 T-lymphocyte imbalance, low CD4-positive T-lymphocyte counts and Kaposi's sarcoma (KS) without HIV infection is reported. Cytomegalovirus inclusions were identified in the nuclei of some KS spindle and endothelial cells. It is known that KS has a high prevalence in AIDS patients and has occasionally been associated with other causes of immunosuppression. In previous studies, coexisting KS and thymoma were related to myasthenia gravis, corticosteroid treatment and excess CD8-positive T lymphocyte counts. More recently an imbalance between CD4 and CD8 positive T lymphocytes has been identified in association with thymoma. The present case suggests that there may be a relationship between thymoma, CD4-positive lymphopenia, and KS. PMID- 9343331 TI - Congenital oligodendroglioma: a case report of a 34th-gestational week fetus with immunohistochemical study and review of the literature. AB - A case of congenital oligodendroglioma occurring in a 34th-gestational week fetus is reported. The tumor was necrotic, hemorrhagic, and gelatinous. It covered the basal part of the brain, and almost the entire cerebellum was replaced by the tumor. The tumor cells had small, round, hyperchromatic nuclei and watery clear cytoplasm, and were arranged in a paved or alveolar pattern. Immunohistochemically, S100 protein, myelin-basic protein, neuron-specific enolase and Leu 7 were weakly positive for the cytoplasm, but glial fibrilliary acidic protein, synaptophysin, neurofilament, desmin, and vimentin were negative. Many tumor cell nuclei were positive for mutant p53 protein, and the labeling index was 85%. But there was no genetic alteration in exons 4 to 9 of p53 gene from the peripheral blood. The apoptosis index was 1.5%. Considering the p53 labeling index and the apoptosis index together, this congenital oligodendroglioma may be regarded as potentially malignant despite the benign morphological features. PMID- 9343332 TI - "Cervical cytology: perspectives from both sides of the Atlantic". PMID- 9343333 TI - Pap smears are financially undervalued in the United States. PMID- 9343334 TI - The cervix cancer screening program in the UK. PMID- 9343335 TI - Immunophenotypic characterization of feline Langerhans cells. AB - To carry out the characterization of feline Langerhans cells (LC), first described in 1994, we used a panel of monoclonal antibodies (MAb) known to react with human, canine and feline leukocyte membrane antigens (Ag). The immunolabeling was performed, at light microscope level, on frozen sections of feline skin and labial mucosa using an avidin-biotin-peroxidase technique, and at electron microscope level on epidermal cell suspensions using an immunogold technique. Out of the 52 MAb tested, six labeled basal or suprabasal DC cells in the frozen sections, either in epidermis or lip epithelium: MHM23 (anti-human CD18), CVS20 and vpg3 (respectively anti-canine and feline-major histocompatibility complex class II molecules), vpg5 (anti-feline leukocytes), vpg39 (anti-feline CD4) and Fel5F4 (anti-feline CD1a). These six MAb were used on suspensions, and labeled cells which showed no desmosomes or melanosomes, but contained 'zipper-like' structures similar to Birbeck granules (BG) in their cytoplasm, revealing they were LC. Consequently, feline LC are CD18-positive (CD18+), major histocompatibility complex class II-positive (Class II+), CD1a positive (CD1a+), vpg5-positive (vg5+) and CD4-positive (CD4+). This immunophenotypic and ultrastructural characterization demonstrates that feline LC share many characteristics with their human counterparts, a fact that will allow us to study the role of feline LC in certain feline diseases such as Feline Immunodeficiency Virus (FIV) infection, since it has been shown that human LC cells are HIV-permissive, and to establish an animal model for human AIDS. PMID- 9343336 TI - Elevated interleukin 6 activity in aqueous humor of cats with uveitis. AB - The purpose of this study was to assess the role of interleukin 6 (IL-6) in feline uveitis by measuring IL-6 activity in the serum and aqueous humor of cats. Serum and aqueous humor was collected from clinically normal, random source cats (n = 10); clinically normal, specific-pathogen free cats experimentally inoculated with Toxoplasma gondii strain ME49 and sampled sequentially for 20 months (n = 4); and client-owned cats with uveitis (n = 27). Interleukin 6 activity was measured in each sample. Client-owned cats with uveitis were also evaluated for evidence of present or prior exposure to T. gondii, feline leukemia virus, feline immunodeficiency virus, and feline coronaviruses. Interleukin 6 activity was non-detectable or low in serum from cats of each group. Interleukin 6 activity was not detected in aqueous humor of clinically normal cats. Interleukin 6 activity was detected in 22/27 (81.5%) aqueous humor samples from cats with uveitis, with a range of 28.9 U ml(-1)-15702.9 U ml(-1) (mean = 1911.9 U ml[-1], SD = 3946.7 U ml[-1]). Serologic evidence of exposure to T gondii, feline immunodeficiency virus, feline leukemia virus, or a coronavirus was present in 21/27 (77.8%) cats with uveitis. Interleukin 6 was detected in the aqueous humor of 18/21 (85.7%) and 3/6 (50%) of the cats with and without serologic evidence of exposure to one to the infectious diseases, respectively. Statistically significant increases in mean IL-6 activity in aqueous humor were found for cats with any evidence of infection with T. gondii, for cats with T. gondii antigen in aqueous humor and for cats with coronavirus antibody titers > or = 1:100. Aqueous humor IL-6 activity was greater than corresponding serum IL-6 activity in 21/27 cats. These results show that IL-6 is produced intraocularly in some cats with uveitis and that IL-6 may be a mediator of uveitis in cats. PMID- 9343337 TI - Induction of procoagulant activity in virus infected bovine alveolar macrophages and the effect of lipopolysaccharide. AB - Three viruses known to be associated with the bovine respiratory disease complex were evaluated in vitro for potential impact upon the procoagulant activity (PCA) of bovine alveolar macrophages (bAM). Cultures of bAM were inoculated with bovine parainfluenza virus Type 3 (PI-3), cytopathic bovine viral diarrhea virus (cpBVDV), non-cytopathic BVDV (ncpBVDV), or bovine herpes virus Type 1 (BHV-1) and incubated for several time periods (24, 48, 72, 96 h). BAM were then exposed to E. coli lipopolysaccharide (LPS), or LPS with bovine serum. The amount of PCA expressed was quantified using a chromogenic assay. Viral inoculation increased bAM expression of PCA (P < 0.01). The increase in PCA expression was larger at higher rates of viral inoculation (P < 0.01). LPS enhanced PCA expression by bAM at low rates of viral inoculation (P < 0.01). The effect of LPS-serum treatment was greater than the LPS alone (P < 0.01). At high rates of viral inoculation, LPS had no enhancing effect on PCA expression. The effect of LPS on virus inoculated bAM varied with virus type, rate of inoculation, and duration of virus exposure (P < 0.01). The results suggest that these four viruses initiate the production of PCA by bAM independently of LPS. In the field situation, an initial viral infection may induce fibrin deposition in the pulmonary alveoli prior to the establishment of a secondary gram negative bacterial infection. PMID- 9343338 TI - Antigen-specific immune responses in cattle with inherited beta2-integrin deficiency. AB - The significance of beta2-integrins for the generation of antigen-specific immune responses in vivo was studied employing the bovine model of beta2-integrin deficiency. To that end four cattle with bovine leukocyte adhesion deficiency (BLAD) and healthy age-matched controls were immunized with tetanus toxoid (TT) and rabies virus (RV) vaccines three times in monthly intervals. In addition, two animals with BLAD and three controls received a fourth vaccination 8 months after the start of the study. Proliferative responses of peripheral blood mononuclear cells (PBMC) to the antigens TT and RV as well as specific serum immunoglobulin G (IgG) titers were determined in intervals for up to 10 months after primary vaccination. Proliferative responses of PBMC to TT and RV were substantially lower in cattle with BLAD than in controls, although PBMC from cattle with BLAD were shown to have the capacity to proliferate in the response to the mitogen concanavalin A. Occurrence of antigen-specific IgG titers was delayed and they were considerably lower in cattle with BLAD compared to controls. Finally, treatment of TT- and RV-stimulated PBMC from an immunized control with different concentrations of the anti-CD18 monoclonal antibody R15.7 resulted in a dose dependent inhibition of lymphocyte proliferation to almost 100%. The results of the present study show that beta2-integrin deficiency leads to delayedand severely impaired immune responsiveness in vivo. The observations that antibody production, although considerably delayed and impaired, does occur and that apparently class-switching takes place in BLAD indicate T-cell reactivity in vivo. PMID- 9343340 TI - Cytokine profile induced by a primary infection with Ostertagia ostertagi in cattle. AB - Changes that occur in the local draining lymph nodes including, changes in cell surface markers and cytokine gene expression were studied over the first 4 weeks of a primary, Ostertagia ostertagi infection of the abomasum. Cells recovered from the abomasal lymph nodes (ABLN) after infection showed a decrease in the percentage of CD3+ cells, and an increase in the percentage of IgM+ cells and cells bearing the TcR1 marker. These changes were coincident with an increase in the proportion of activated cells (II-2R). Analysis of mitogen-stimulated ABLN cells by RNase protection assay (RPA) showed a dramatic reduction in IL-2 and IFN gamma transcription after infection. In addition, analysis of unstimulated ABLN cells by competitive RT-PCR showed a similar decrease in demonstrable levels of IL-2 mRNA, but IL-10, IL-4 and IFN-gamma mRNA levels were elevated. PMID- 9343339 TI - Memory and CD8+ are the predominant bovine bronchoalveolar lymphocyte phenotypes. AB - Bovine lymphocytes obtained by bronchoalveolar lavage (BAL) of healthy calves were simultaneously analyzed and compared to peripheral blood lymphocytes using monoclonal antibodies specific for bovine leukocyte differentiation antigens. Phenotypic differences were observed between bronchoalveolar and peripheral blood T-lymphocyte subpopulations, demonstrating selective lymphocyte migration to the bovine lung. The bronchoalveolar and peripheral blood T-lymphocyte populations, defined by expression of CD2, were similar, but bronchoalveolar T lymphocytes were predominately CD8+ while peripheral blood T cells were predominately CD4+. In addition, memory lymphocytes, characterized by low expression of CD45R and activated lymphocytes (CD25+), were found in significantly higher proportions in the bronchoalveolar compartment. The proportion of gammadelta T lymphocytes was, however, significantly higher in peripheral blood. B cells were observed in similar proportions in the bronchoalveolar compartment and peripheral blood. PMID- 9343341 TI - Bronchoalveolar cellular responses of goats following infections with Muellerius capillaris (Protostrongylidae, Nematoda). AB - The development of Muellerius capillaris in the lung of goats was associated with marked tissue damage and pronounced a cellular reaction. Using broncho-alveolar lavage, the time course of the cellular responses was studied following primary and secondary infection. During the primary infection, there was a biphasic increase in total broncho-alveolar leucocytes (an average of 294.0 +/- 137.0 cells microl[-1]) and in the absolute number of macrophages (182.0 +/- 82.0 cells ul[-1]), lymphocytes (68.5 +/- 35.0 cells microl[-1]), eosinophils (35.3 +/- 16.4 cells microl[-1]) and neutrophils (10.9 +/- 8.7 cell microl[-1]). The lung tissue reaction against worms consisted of a mild infiltration of inflammatory cells. The secondary infection resulted in significant changes in the pulmonary tissue characterised by severe inflammation, leading to widespread granulomatous formation throughout the parenchyma, hyperplasia of cells Type II and a leucocytosis in the broncho-alveolar fluids, with an anamnestic-like response by all cell types. The overall average of the total leucocytes, macrophages, lymphocytes, eosinophils and neutrophils was 529.3 +/- 347.4; 265.4 +/- 148.1; 127.3 +/- 100; 125.4 +/- 100.1 and 14.0 +/- 8.7 cells microl(-1), respectively. Secondary infection also resulted in 56% reduction of worms established in the lungs and 72.3% of L1 larval production. These data suggest that the broncho alveolar leucocyte response to infection has an immunological basis and that the alveolar macrophages, neutrophils, eosinophils, and lymphocytes may play a significant role in lung resistance against protostrongylid nematodes. PMID- 9343342 TI - Small is powerful: recollections of a microbiologist and oceanographer. PMID- 9343343 TI - Xenorhabdus and Photorhabdus spp.: bugs that kill bugs. AB - Xenorhabdus and Photorhabdus spp. are gram negative gamma proteobacteria that form entomopathogenic symbioses with soil nematodes. They undergo a complex life cycle that involves a symbiotic stage, in which the bacteria are carried in the gut of the nematodes, and a pathogenic stage, in which susceptible insect prey are killed by the combined action of the nematode and the bacteria. Both bacteria produce antibiotics, intracellular protein crystals, and numerous other products. These traits change in phase variants, which arise when the bacteria are maintained under stationary phase conditions in the laboratory. Molecular biological studies suggest that Xenorhabdus and Photorhabdus spp. may serve as valuable model systems for studying signal transduction and transcriptional and posttranscriptional regulation of gene expression. Such studies also indicate that these bacterial groups, which had been previously considered to be very similar, may actually be quite different at the molecular level. PMID- 9343344 TI - Molecular genetics of sulfur assimilation in filamentous fungi and yeast. AB - The filamentous fungi Aspergillus nidulans and Neurospora crassa and the yeast Saccharomyces cerevisiae each possess a global regulatory circuit that controls the expression of permeases and enzymes that function both in the acquisition of sulfur from the environment and in its assimilation. Control of the structural genes that specify an array of enzymes that catalyze reactions of sulfur metabolism occurs at the transcriptional level and involves both positive-acting and negative-acting regulatory factors. Positive trans-acting regulatory proteins that contain a basic region, leucine zipper-DNA binding domain, are found in Neurospora and yeast. Each of these fungi contain a sulfur regulatory protein of the beta-transducin family that acts in a negative fashion to control gene expression. Sulfur regulation in yeast also involves the general DNA binding protein, centromere binding factor I. Sulfate uptake is a highly regulated step and appears to occur in fungi, plants, and mammals via a family of related transporter proteins. Recent developments have provided new insight into the nature and control of the enzymes ATP sulfurylase and APS kinase, which catalyze the early steps of sulfate assimilation, and of the Aspergillus enzyme, cysteine synthase, which produces cysteine from O-acetylserine. PMID- 9343345 TI - Hemoglobin metabolism in the malaria parasite Plasmodium falciparum. AB - Hemoglobin degradation in intraerythrocytic malaria parasites is a vast process that occurs in an acidic digestive vacuole. Proteases that participate in this catabolic pathway have been defined. Studies of protease biosynthesis have revealed unusual targeting and activation mechanisms. Oxygen radicals and heme are released during proteolysis and must be detoxified by dismutation and polymerization, respectively. The quinoline antimalarials appear to act by preventing sequestration of this toxic heme. Understanding the disposition of hemoglobin has allowed identification of essential processes and metabolic weakpoints that can be exploited to combat this scourge of mankind. PMID- 9343346 TI - Getting started: regulating the initiation of DNA replication in yeast. AB - Initiation of DNA replication in yeast appears to operate through a two-step process. The first step occurs at the end of mitosis in the previous cell cycle, where, following the decrease in B cyclin-dependent kinase activity, an extended protein complex called the prereplicative complex (pre-RC) forms over the origin of replication. This complex is dependent on the association of the Cdc6 protein with the Origin Recognition Complex (ORC) and appears concomitantly with the nuclear entry of members of the Mcm family of proteins. The second step is dependent upon the cell passing through a G1 decision point called Start. If the environmental conditions are favorable, and the cells reach a critical size, then there is a rise in G1 cyclin-dependent kinase activity, which leads to the activation of downstream protein kinases; the protein kinases are, in turn, required for triggering initiation from the preformed initiation complexes. These protein kinases, Dbf4-Cdc7 and Clb5/6(B-cyclin)-Cdc28, are thought to phosphorylate targets within the pre-RC. The subsequent rise in B cyclin protein kinase activity following Start not only triggers origin firing, but also inhibits the formation of new pre-RCs, which ensures that there is only one S phase in each cell cycle. The destruction of B-cyclin protein kinase activity at the end of the cell cycle potentiates the formation of new pre-RCs-resetting origins for the next S phase. PMID- 9343347 TI - RNA virus mutations and fitness for survival. AB - RNA viruses exploit all known mechanisms of genetic variation to ensure their survival. Distinctive features of RNA virus replication include high mutation rates, high yields, and short replication times. As a consequence, RNA viruses replicate as complex and dynamic mutant swarms, called viral quasispecies. Mutation rates at defined genomic sites are affected by the nucleotide sequence context on the template molecule as well as by environmental factors. In vitro hypermutation reactions offer a means to explore the functional sequence space of nucleic acids and proteins. The evolution of a viral quasispecies is extremely dependent on the population size of the virus that is involved in the infections. Repeated bottleneck events lead to average fitness losses, with viruses that harbor unusual, deleterious mutations. In contrast, large population passages result in rapid fitness gains, much larger than those so far scored for cellular organisms. Fitness gains in one environment often lead to fitness losses in an alternative environment. An important challenge in RNA virus evolution research is the assignment of phenotypic traits to specific mutations. Different constellations of mutations may be associated with a similar biological behavior. In addition, recent evidence suggests the existence of critical thresholds for the expression of phenotypic traits. Epidemiological as well as functional and structural studies suggest that RNA viruses can tolerate restricted types and numbers of mutations during any specific time point during their evolution. Viruses occupy only a tiny portion of their potential sequence space. Such limited tolerance to mutations may open new avenues for combating viral infections. PMID- 9343348 TI - Making and breaking disulfide bonds. AB - It is now well established that protein folding requires the assistance of folding helpers in vivo. The formation or isomerization of disulfide bonds in proteins is a slow process requiring catalysis. In nascent polypeptide chains the cysteine residues are in the thiol form. The formation of the disulfide bonds usually occurs simultaneously with the folding of the polypeptide, which means in the endoplasmic reticulum of eukaryotes or in the periplasm of Gram-negative bacteria. In prokaryotes, the existence of redox proteins involved in the formation of disulfide bonds containing proteins has recently been revealed in the periplasm. The discovery of these redox proteins through various genetic approaches will be summarized, as well as the most recent insights regarding their biochemical and biological activities. PMID- 9343349 TI - Against all odds: the survival strategies of Deinococcus radiodurans. AB - Bacteria of the genus Deinococcus exhibit an extraordinary ability to withstand the lethal and mutagenic effects of DNA damaging agents-particularly the effects of ionizing radiation. These bacteria are the most DNA damage-tolerant organisms ever identified. Relatively little is known about the biochemical basis for this phenomenon; however, available evidence indicates that efficient repair of DNA damage is, in large part, responsible for the deinococci's radioresistance. Obviously, an explanation of the deinococci's DNA damage tolerance cannot be developed solely on the basis of the DNA repair strategies of more radiosensitive organisms. The deinococci's capacity to survive DNA damage suggests that (a) they employ repair mechanisms that are fundamentally different from other prokaryotes, or that (b) they have the ability to potentiate the effectiveness of the conventional complement of DNA repair proteins. An argument is made for the latter alternative. PMID- 9343350 TI - Genetics of the rotaviruses. AB - Genetic analyses have contributed significantly to our understanding of the biology of the rotaviruses. The distinguishing feature of the virus is a genome consisting of 11 segments of double-stranded RNA. The segmented nature of the genome allows reassortment of genome segments during mixed infections, which is the major distinguishing feature of rotavirus genetics. Reassortment has been a powerful tool for mapping viral mutations and other determinants of biological phenotypes to specific genome segments. However, more detailed genetic analysis of rotaviruses is currently limited by the inability to perform reverse genetics. Development of a reverse genetic system will facilitate analysis of the molecular mechanisms involved in various genetic, biochemical, and biological phenomena of the virus. PMID- 9343351 TI - Intracellular antibodies (intrabodies) for gene therapy of infectious diseases. AB - Intracellular antibodies (intrabodies) represent a new class of neutralizing molecules with a potential use in gene therapy. Intrabodies are engineered single chain antibodies in which the variable domain of the heavy chain is joined to the variable domain of the light chain through a peptide linker, preserving the affinity of the parent antibody. Intrabodies are expressed inside cells and directed to different subcellular compartments where they can exert their function more effectively. The effects of intrabodies have been investigated using structural, regulatory, and enzymatic proteins of the human immunodeficiency virus (HIV-1) as targets. These intrabodies have demonstrated their versatility by controlling early as well as late events of the viral life cycle. In this article, we review studies of the use of intrabodies as research tools and therapeutic agents against HIV-1. PMID- 9343352 TI - Microbial aldolases and transketolases: new biocatalytic approaches to simple and complex sugars. AB - Enzymes have become exceedingly valuable tools in organic synthesis as the reactions they catalyze generally proceed under mild conditions and in high stereo- and regioselectivity. Advances in microbiology and genetic engineering have greatly increased the availability of various enzymes. One of the most useful applications of enzyme-catalyzed chemical transformations is in the synthesis of water-soluble, polyfunctional organic molecules such as carbohydrates. As the pivotal roles that carbohydrates play in biological processes become more evident, access to these compounds becomes increasingly important. This review gives a brief overview of the use of aldolases and transketolases in the synthesis of sugars, sugar analogs, and related compounds. PMID- 9343353 TI - Interaction of antigens and antibodies at mucosal surfaces. AB - Infections often involve the mucosal surfaces of the body, which form a boundary with the outside world. This review focuses on immunoglobulin A (IgA) antibodies because IgA is the principal mucosal antibody class. IgA is synthesized by local plasma cells and has a specific polymeric immunoglobulin receptor-mediated transport mechanism for entry into the secretions. By serving as an external barrier capable of inhibiting attachment of microbes to the luminal surface of the mucosal epithelial lining, IgA antibodies form the first line of immune defense. In addition to this traditional mode of extracellular antibody function, recent evidence suggests that IgA antibodies can also function in a nontraditional fashion by neutralizing viruses intracellularly, if a virus is infecting an epithelial cell through which specific IgA antibody is passing on its way to the secretions. IgA antibodies are also envisaged as providing an internal mucosal barrier beneath the mucosal lining. Antigens intercepted by IgA antibodies here can potentially be ferried through the epithelium and thereby excreted. In addition to the polymeric immunoglobulin receptor on mucosal epithelial cells, IgA antibodies can bind to receptors on a variety of leukocytes and have been shown, in some experimental systems, to be capable of activating the alternative complement pathway, making IgA antibodies potential participants in inflammatory reactions. Although the relationship of IgA antibodies to inflammation is not entirely clear, the bias presented is that IgA is basically noninflammatory, perhaps even anti-inflammatory. According to this view, the major role of the Fc portion of IgA antibodies is to transport IgA across mucosal epithelial cells and not, as in the case of the other classes of antibody, to activate secondary phenomena of the kind that contribute to inflammation. Because of IgA's key role as an initial barrier to infection, much current research in mucosal immunology is directed toward developing new vectors and adjuvants that can provide improved approaches to mucosal vaccines. Finally, because of advances in monoclonal antibody technology, topical application of antibodies to mucosal surfaces has significant potential for preventing and treating infections. PMID- 9343354 TI - Transcriptional control of the Pseudomonas TOL plasmid catabolic operons is achieved through an interplay of host factors and plasmid-encoded regulators. AB - The xyl genes of Pseudomonas putida TOL plasmid that specify catabolism of toluene and xylenes are organized in four transcriptional units: the upper-operon xylUWCAMBN for conversion of toluene/xylenes into benzoate/alkylbenzoates; the meta-operon xylXYZLTEGFJQKIH, which encodes the enzymes for further conversion of these compounds into Krebs cycle intermediates; and xylS and xylR, which are involved in transcriptional control. The XylS and XylR proteins are members of the XylS/AraC and NtrC families, respectively, of transcriptional regulators. The xylS gene is constitutively expressed at a low level from the Ps2 promoter. The XylS protein is activated by interaction with alkylbenzoates, and this active form stimulates transcription from Pm by sigma70- or sigmaS-containing RNA polymerase (the meta loop). The xylR gene is also expressed constitutively. The XylR protein, which in the absence of effectors binds in a nonactive form to target DNA sequences, is activated by aromatic hydrocarbons and ATP; it subsequently undergoes multimerization and structural changes that result in stimulation of transcription from Pu of the upper operon. This latter process is assisted by the IHF protein and mediated by sigma54-containing RNA polymerase. Once activated, the XylR protein also stimulates transcription from the Ps1 promoter of xylS without interfering with expression from Ps2. This process is assisted by the HU protein and is mediated by sigma54-containing RNA polymerase. As a consequence of hyperexpression of the xylS gene, the XylS protein is hyperproduced and stimulates transcription from Pm even in the absence of effectors (the cascade loop). The two sigma54-dependent promoters are additionally subject to global (catabolite repression) control. PMID- 9343355 TI - Sulfur tuft and turkey tail: biosynthesis and biodegradation of organohalogens by Basidiomycetes. AB - Chlorinated aliphatic and aromatic compounds are generally considered to be undesirable xenobiotic pollutants. However, the higher fungi, Basidiomycetes, have a widespread capacity for organohalogen biosynthesis. Adsorbable organic halogens (AOX) and/or low-molecular-weight halogenated compounds are produced by Basidiomycetes of 68 genera from 20 different families. Most of the 81 halogenated metabolites identified from Basidiomycetes to date are chlorinated, although brominated and iodated metabolites have also been described. Two broad categories of Basidiomycete organohalogen metabolites are the halogenated aromatic compounds and the haloaliphatic compounds. Some of these organohalogen metabolites have demonstrable physiological roles as antibiotics and as metabolites involved in lignin degradation. Basidiomycetes produce large amounts of low-molecular-weight organohalogens or adsorbable organic halogens (AOX) when grown on lignocellulosic substrates. In our view, Basidiomycetes, as decomposers of forest litter, are a major source of natural organohalogens in terrestrial environments. Basidiomycetes are also potent degraders of a wide range of chlorinated pollutants, such as bleachery effluent from kraft mills and pentachlorophenol, polychlorinated dioxins, and polychlorinated biphenyls. The extracellular, lignin-degrading enzymes of the Basidiomycetes are involved in the oxidative degradation of chlorophenols and dioxin and can cause reductive dechlorination of halomethanes. There is no clear-cut separation between "polluters" and "clean-uppers" within the Basidiomycetes. Several genera, e.g. Bjerkandera, Hericium, Phlebia, and Trametes, produce significant amounts of chlorinated compounds but are also highly effective in metabolizing or biotransforming chlorinated pollutants. PMID- 9343356 TI - Safe haven: the cell biology of nonfusogenic pathogen vacuoles. AB - Our understanding of both membrane traffic in mammalian cells and the cell biology of infection with intracellular pathogens has increased dramatically in recent years. In this review, we discuss the cell biology of the host-microbe interaction for four intracellular pathogens: Chlamydia spp., Legionella pneumophila, Mycobacterium spp., and the protozoan parasite Toxoplasma gondii. All of these organisms reside in vacuoles inside cells that have restricted fusion with host organelles of the endocytic cascade. Despite this restricted fusion, the vacuoles surrounding each pathogen display novel interactions with other host cell organelles. In addition to the effect of infection on host membrane traffic, we focus on these novel interactions and relate them where possible to nutrient acquisition by the intracellular organisms. PMID- 9343357 TI - Transcription of protein-coding genes in trypanosomes by RNA polymerase I. AB - In eukaryotes, RNA polymerase (pol) II transcribes the protein-coding genes, whereas RNA pol I transcribes the genes that encode the three RNA species of the ribosome [the ribosomal RNAs (rRNAs)] at the nucleolus. Protozoan parasites of the order Kinetoplastida may represent an exception, because pol I can mediate the expression of exogenously introduced protein-coding genes in these single cell organisms. A unique molecular mechanism, which leads to pre-mRNA maturation by trans-splicing, facilitates pol I-mediated protein-coding gene expression in trypanosomes. Trans-splicing adds a capped 39-nucleotide mini-exon, or spliced leader transcript, to the 5' end of the main coding exon posttranscriptionally. In other eukaryotes, the addition of a 5' cap, which is essential for mRNA function, occurs exclusively as a result of RNA pol II-mediated transcription. Given the assumption that cap addition represents the limiting factor, trans splicing may have uncoupled the requirement for RNA pol II-mediated mRNA production. A comparison of the alpha-amanitin sensitivity of transcription in naturally occurring trypanosome protein-coding genes reveals that a unique subset of protein-coding genes-the variant surface glycoprotein (VSG) expression sites and the procyclin or the procyclic acidic repetitive protein (PARP) genes-are transcribed by an RNA polymerase that is resistant to the mushroom toxin alpha amanitin, a characteristic of transcription by RNA pol I. Promoter analysis and a pharmacological characterization of the RNA polymerase that transcribes these genes have strengthened the proposal that the VSG expression sites and the PARP genes represent naturally occurring protein-coding genes that are transcribed by RNA pol I. PMID- 9343358 TI - Synthesis of enantiopure epoxides through biocatalytic approaches. AB - Enantiopure epoxides, as well as their corresponding vicinal diols, are valuable intermediates in fine organic synthesis, in particular for the preparation of biologically active compounds. The necessity of preparing such target molecules in an optically pure form has triggered much research, leading to the emergence of various new methods based on either conventional chemistry or enzymatically catalyzed reactions. In this review, we focus on the biocatalytic approaches, which include direct epoxidation of olefinic double bonds as well as indirect biocatalytic methods, and which allow for the synthesis of these important chiral building blocks in enantiomerically enriched or even enantiopure form. PMID- 9343359 TI - Cell-cell communication in gram-positive bacteria. AB - In gram-positive bacteria, many important processes are controlled by cell-to cell communication, which is mediated by extracellular signal molecules produced by the bacteria. Most of these signaling molecules are peptides or modified peptides. Signal processing, in most cases, involves either transduction across the cytoplasmic membrane or import of the signal and subsequent interaction with intracellular effectors. Concentrations of signal in the nanomolar range or below are frequently sufficient for biological activity. The microbial processes controlled by extracellular signaling include the expression of virulence factors, the expression of gene transfer functions, and the production of antibiotics. PMID- 9343360 TI - Regulators of apoptosis on the road to persistent alphavirus infection. AB - Alphavirus infection can trigger the host cell to activate its genetically programmed cell death pathway, leading to the morphological features of apoptosis. The ability to activate this death pathway is dependent on both viral and cellular determinants. The more virulent strains of alphavirus induce apoptosis with increased efficiency both in animal models and in some cultured cells. Although the immune system clearly plays a central role in clearing virus, the importance of other cellular factors in determining the outcome of virus infections are evident from the observation that mature neurons are better able to resist alphavirus-induced apoptosis than immature neurons are, both in culture and in mouse brains. These findings are consistent with the age-dependent susceptibility to disease seen in animals. Cellular genes that are known to regulate the cell death pathway can modulate the outcome of alphavirus infection in cultured cells and perhaps in animals. The cellular bax and bak genes, which are known to accelerate cell death, also accelerate virus-induced apoptosis. In contrast, inhibitors of apoptotic cell death such as bcl-2 suppress virus-induced apoptosis, which can facilitate a persistent virus infection. Thus, the balance of cellular factors that regulate cell death may be critical in virus infections. Additional viral factors also contribute to this balance. The more virulent strains of alphavirus have acquired the ability to induce apoptosis in mature neurons, while mature neurons are resistant to cell death upon infection with less virulent strains. Here we discuss a variety of cellular and viral factors that modulate the outcome of virus infection. PMID- 9343361 TI - Clues and consequences of DNA bending in transcription. AB - This review attempts to substantiate the notion that nonlinear DNA structures allow prokaryotic cells to evolve complex signal integration devices that, to some extent, parallel the transduction cascades employed by higher organisms to control cell growth and differentiation. Regulatory cascades allow the possibility of inserting additional checks, either positive or negative, in every step of the process. In this context, the major consequence of DNA bending in transcription is that promoter geometry becomes a key regulatory element. By using DNA bending, bacteria afford multiple metabolic control levels simply through alteration of promoter architecture, so that positive signals favor an optimal constellation of protein-protein and protein-DNA contacts required for activation. Additional effects of regulated DNA bending in prokaryotic promoters include the amplification and translation of small physiological signals into major transcriptional responses and the control of promoter specificity for cognate regulators. PMID- 9343362 TI - Genetics of eubacterial carotenoid biosynthesis: a colorful tale. AB - Carotenoids represent one of the most widely distributed and structurally diverse classes of natural pigments, with important functions in photosynthesis, nutrition, and protection against photooxidative damage. In the eubacterial community, yellow, orange, and red carotenoids are produced by anoxygenic photosynthetic bacteria, cyanobacteria, and certain species of nonphotosynthetic bacteria. Many eukaryotes, including all algae and plants, as well as some fungi, also synthesize these pigments. In noncarotenogenic organisms, such as mammals, birds, amphibians, fish, crustaceans, and insects, dietary carotenoids and their metabolites also serve important biological roles. Within the last decade, major advances have been made in the elucidation of the molecular genetics, the biochemistry, and the regulation of eubacterial carotenoid biosynthesis. These developments have important implications for eukaryotes, and they make increasingly attractive the genetic manipulation of carotenoid content for biotechnological purposes. PMID- 9343363 TI - Enzyme activity alteration by cadmium administration to rats: the possibility of iron involvement in lipid peroxidation. AB - The specific activities of D-3-hydroxybutyrate dehydrogenase (BDH) and glutamate dehydrogenase (GDH) are reduced in the liver and kidney of rats intoxicated with 2.5 mg Cd/kg body wt and sacrificed after 24 h; conversely ketone-body concentration is strongly increased in both of these organs and blood. In the same animals a great stimulation of antioxidant enzymes glutathione reductase and glutathione peroxidase occurs. The prooxidant state induced by cadmium in liver mitochondria and microsomes is unaffected by superoxide dismutase, catalase, or mannitol, whereas it is completely blocked by vitamin E thus excluding the involvement of reactive oxygen species in this process. The mechanism by which cadmium induces lipid peroxidation has been investigated by measuring the effect of this metal on liposomes. Ninety-minute treatment of liposomes with CdCl2 does not induce any lipid peroxidation. In contrast, Fe2+ ions under the same conditions cause strong liposome peroxidation. It has also been observed that cadmium promotes a time-dependent iron release from biological membranes. When lipid peroxidation is induced by a low concentration (5 microM) of FeCl2, in place of CdCl2, the characteristics of this process and the sensitivity to the various antioxidants used are similar to those observed with Cd. From these results we conclude that the prooxidative effect of cadmium is an indirect one since it is mediated by iron. With regard to the inhibitory effect on BDH and GDH following cadmium intoxication, it does not appear to be imputable to lipid peroxidation since in vitro investigations indicate that the presence of vitamin E does not remove the inhibition at all. PMID- 9343364 TI - Release of iron from ferritin storage by redox cycling of stilbene and steroid estrogen metabolites: a mechanism of induction of free radical damage by estrogen. AB - Estrogens induce hydroxyl radical-mediated DNA and protein damage and lipid peroxidation. As part of a study of the mechanism of hydroxyl radical generation by estrogens, we investigated the in vitro mobilization of Fe2+ from ferritin by redox cycling of the stilbene or steroid estrogen metabolites diethylstilbestrol 4',4"-quinone (DESQ), equilenin-3,4-quinone (EQ), or estrone-3,4-quinone (3,4EQ). Aerobic cytochrome P450 reductase-mediated redox cycling of 35.50 microM DESQ, 0.35 microM EQ, or 3.55 microM 3,4EQ increased the reduction of succinoylated cytochrome c, a measure of superoxide radical formation, by 19-20% over control values (24.5+/-0.3 microM) in the absence of estrogen quinone substrate. Rates of Fe2+ release from horse spleen ferritin by cytochrome P450 reductase-mediated redox cycling of 35.50 microM DESQ, 0.35 microM EQ, or 3.55 microM 3,4EQ were 94.4+/-0.6, 117.2+/-9.4, or 137.7+/-19.9 pmol Fe2+/min, respectively, compared to 67.3 + 2.3 pmol Fe2+/min in the absence of estrogen substrates. Redox cycling of 35.5 microM DESQ, EQ, or 3,4EQ mediated by microsomes of hamster kidney, a target organ of estrogen-induced carcinogenesis, released 511+/-30.10, 516.91+/-22.90, or 410.27+/-28.49 pmol Fe2+/min, respectively. Corresponding values with microsomes of hamster liver, where tumors do not develop by estrogen treatment, were 272.27+/-43.10, 222.25+/-21.78, or 91.36+/-8.54 pmol Fe2-/min, respectively. Diethylstilbestrol, equilenin, and 4-hydroxyestrone do not induce detectable iron release from ferritin under these conditions. The cytochrome P450 reductase mediated redox cycling of DESQ, EQ, or 3,4EQ in the presence of iron resulted in the hydroxylation of benzoic acid by hydroxyl radical attack. These data demonstrate that redox cycling of estrogen metabolites releases Fe2+ from ferritin, which in turn generates hydroxyl radicals by a Fenton reaction. This estrogen-induced hydroxyl radical damage may contribute to tumor initiation in hormone target tissues, including breast cancer. PMID- 9343366 TI - Interactions of a high mobility group-like protein with human mitochondrial DNA. AB - A number of nuclear-encoded proteins are known to bind to the mitochondrial genome and may be involved in mitochondrial replication and in mitochondrial diseases. Mitochondrial diseases such as Kearns-Sayre syndrome (KSS) are flanked by common direct repeats. To study protein binding to these mitochondria DNA regions we used gel mobility shift binding assays. Proteins present in nuclear or in mitochondrial extracts from normal or KSS-derived fibroblasts bound to a 280 bp mitochondrial DNA fragment encompassing a deletion breakpoint in the mitochondrial genome. In addition, nuclear and mitochondrial protein bound to a nearby surrounding fragment and to a synthetic oligonucleotide with a related consensus DNA sequence. Southwestern blot analysis showed that the DNA binding ability resided in a 35-kDa protein. The amino terminal sequence of the 35-kDa protein was very similar to human high mobility group proteins. These results suggest that this protein may be involved in mitochondrial DNA deletions. PMID- 9343365 TI - Activation of liver mitochondrial phospholipase A2 by superoxide. AB - Mitochondrial damage is one of the prominent features of cell injury during oxidative stress and altered mitochondrial lipids may contribute to this damage. Lipid changes were observed when liver mitochondria were exposed to superoxide generating systems. Phosphatidylcholine and phosphatidylethanolamine contents were decreased with simultaneous formation of lysophospholipids when exposed to superoxide. Among the neutral lipids there was an increase in the level of free fatty acids. This alteration in lipid composition could be prevented by the simultaneous presence of superoxide dismutase or phospholipase A2 (PLA2) inhibitors. H2O2 did not have any effect on liver mitochondrial PLA2. This suggests that superoxide anion stimulates phospholipase A2 which is prevented by superoxide scavenging agents and PLA2 inhibitors. The products of phospholipase A2 are membrane-damaging agents which may be responsible for mitochondrial damage seen during oxidative stress. PMID- 9343367 TI - Interaction of spermine with dimyristoyl-L-alpha-phosphatidyl-DL-glycerol multilamellar liposomes. AB - Polycationic spermine interacts with the negative phosphate group of dimyristoylphosphatidylglycerol multilamellar liposomes, forming a positively charged shell around the vesicle surface. An association constant of (2.15+/ 0.45) x 10(3) M(-1) between spermine and the phospholipid groups in liposomes has been evaluated by a new and rapid enzymatic method. ESR spectra show that the effects of this polycation on liposomes are substantially different from those of cations like Ca2+ and Mg2+ and confirm the ability of spermine to induce liposome aggregation and not fusion. PMID- 9343368 TI - In vitro photoinactivation of catalase isoforms from cotyledons of sunflower (Helianthus annuus L.). AB - Catalase (EC 1.11.1.6) isoforms CAT 2 through CAT 8 were purified from peroxisomes of sunflower (Helianthus annuus L.) cotyledons and photoinactivated in vitro. Action and absorbance spectra between 380 and 727 nm wavelength showed most prominent maxima at 405 nm suggesting an inactivation mediated by light absorption of heme groups. First order kinetics of inactivation were observed for CAT 6 through CAT 8 (isoform group B), which are composed of four 55-kDa subunits. Inactivation constants ki depended on photon fluence rates in the studied range between 8.3 and 660 microE m(-2) s(-1). The maximal value of ki was about 4.0 h(-1), corresponding to a half-life of about 10 min. Heme groups and 55 kDa apoprotein moieties of group B isoforms were degraded during irradiation, but both degradation processes occurred at rates lower than those of inactivation. Quantitative evaluations contradicted the view that photoinactivation was caused by destruction or dissociation of heme but suggested apoprotein damage leading to the loss of activity. Group A isoforms CAT 2 through CAT 5, containing both 55- and 59-kDa subunits, were less photosensitive than the isoforms of group B. In addition, irradiated group A isoforms reached a low plateau of residual activity, whereas group B isoforms were inactivated completely. The 59-kDa subunits in group A isoforms were much more resistant to photodegradation than the 55-kDa subunits of group B isoforms and also much more resistant than their own 55-kDa cosubunits. Results presented here are compared with catalase photoinactivation and turnover in vivo and discussed with respect to a physiological significance of catalase isoforms in plant peroxisomes. PMID- 9343369 TI - Nitrobenzimidazoles as substrates for DT-diaphorase and redox cycling compounds: their enzymatic reactions and cytotoxicity. AB - We have synthesized a number of nitrobenzimidazoles containing nitro groups in the benzene ring and found that they acted as relatively efficient substrates for rat liver DT-diaphorase (EC 1.6.99.2), their reactivity exceeding reactivities of nitrofurans and nitrobenzenes. Nitrobenzimidazoles were competitive with NADPH inhibitors of DT-diaphorase in menadione reductase reactions, their inhibition constant being unchanged in the presence of dicumarol and being increased in the presence of 2',5'-ADP. These data indicate that the poor reactivity of nitrobenzimidazoles and other nitroaromatics in comparison to quinones could be determined by their binding in the adenosine-phosphate binding region of the NADPH-binding site, whereas quinones bind at the nicotinamide-binding pocket at the vicinity of FAD of DT-diaphorase. The reduction of 4,5,6-trinitrobenzimidazol 2-one by DT-diaphorase most probably involves reduction of 5-nitro group to 5 nitroso or 5-hydroxylamine derivative at the initial step. A certain parallelism existed between reactivities of nitrobenzimidazoles toward DT-diaphorase and their reactivities in single-electron reduction by Anabaena ferredoxin:NADP+ reductase (EC 1.18.1.2) and Saccharomyces cerevisiae flavocytochrome b2 (EC 1.1.2.3), the latter being determined by electronic factors. However, we suppose that the relatively high reactivity of polinitrobenzimidazoles toward DT diaphorase was due not only to electronic effects, but also to a sterical crowding of nitrogroups by each other. The toxicity of nitrobenzimidazoles to bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) with a moderate amount of DT-diaphorase (260 U/mg protein) is partly prevented by dicumarol. That points out to partial determination of nitrobenzimidazole cytotoxicity by their reduction by DT-diaphorase. Another important factor of nitrobenzimidazole toxicity to this cell line was oxidative stress, catalyzed by single-electron transferring enzymes. PMID- 9343370 TI - Structural investigation of Tet repressor loop 154-167: a time-resolved fluorescence study of three single Trp mutants. AB - We have studied the time-resolved fluorescence of three engineered Tet repressor (TetR) mutants bearing a single Trp residue at positions 162, 163, and 165 in the C-terminal part of the loop joining helices 8 and 9. Detailed analysis indicates that, at 20 degrees, the fluorescence decay of each Trp can be described as the sum of three exponential components with lifetimes in the 1-, 3-, and 6-ns range. Emission wavelength and temperature dependence studies are consistent with a model in which these components are due to the existence of three classes of Trp residues non-interconverting on the nanosecond timescale. Within the framework of the rotamer model, the weak temperature dependence of the lifetimes strongly suggests that the secondary structure of the loop, at least in the 162-165 range, is not altered with temperature. The equilibrium between the rotamers is characterized by an enthalpy-entropy compensation effect which strongly suggests the involvement of background structural regions of TetR in the thermodynamics of the process. The very high deltaH degrees and TdeltaS degrees observed (up to 18 kcal/ mol) should reflect the temperature-dependent conformational change of a large part of the protein which would alter the rotamer distribution of the Trp residues. Taken together, our results are consistent with the existence of (at least) two conformations of the loop and suggest a model for loop motion. PMID- 9343371 TI - The role of NAD(P)H:quinone oxidoreductase in quinone-mediated p21 induction in human colon carcinoma cells. AB - This study examines the role of NAD(P)H:quinone acceptor oxidoreductase (NQOR) (EC 1.6.99.2) in the metabolism of aziridinylbenzoquinones and the ensuing formation of reactive oxygen species in the induction of the cell cycle inhibitor p21 (WAF1, Cip1, or sdi1) in human colon carcinoma cells. The aziridinylbenzoquinones used were 2,5-diaziridinyl-1,4-benzoquinone (DZQ) and 2,5 bis(carboethoxyamino)-3,6-diaziridinyl-1,4-benzoquinone (AZQ). The cell lines used in this study, BE and HT29 human colon carcinoma cell lines, are devoid of and overexpress NQOR activity, respectively. The rate of reduction of the above quinones in BE cells proceeded at similar rates (approximately 170 nmol/min/ mg protein) and, expectedly, it was not affected by the NQOR inhibitor, dicumarol. The metabolism of DZQ in HT29 cells was largely accomplished by NQOR (approximately 94%), whereas that of AZQ was accomplished by dicumarol insensitive reductases. The metabolism of DZQ in HT29 cells was accompanied by H2O2 formation, which was approximately 10-fold higher than that ensuing from the activation of AZQ. In agreement with these data, the production of H2O2 during the activation of DZQ by purified NQOR was approximately 10-fold higher than that of AZQ. The formation of H2O2 during the metabolism of aziridinylbenzoquinones in BE cells was 24- to 57-fold lower than that in HT29 cells. At variance with HT29 cells, H2O2 formation by BE cells was insensitive to the catalase inhibitor sodium azide. The bioactivation of AZQ and DZQ in BE cells yielded O2.- and HO. as detected by spin trapping/EPR, the intensity of the former adduct being approximately 2-fold higher than that of the latter. These signals were insensitive to dicumarol. The metabolism of DZQ in HT29 cells yielded mainly HO. and a modest contribution of O2.- (ratio HO./O2.- approximately 10), whereas that of AZQ yielded a HO./O2.- approximately 2. The effect of dicumarol on the free radical pattern obtained during DZQ metabolism resulted in a strong inhibition (80%) of HO. production and a substantial increase of O2.- generation. The metabolism of DZQ and AZQ in BE cells was associated with a significant increase of p21 mRNA levels; the former quinone was approximately 2-fold more efficient than the latter. DZQ metabolism in HT29 cells led to an increase of p21 mRNA levels 15-fold higher than that observed with AZQ activation. Dicumarol did not inhibit p21 induction associated with the metabolism of DZQ in the NQOR-deficient BE cells, whereas the inhibitor decreased p21 induction in HT29 cells by approximately 30%. This modest inhibition is likely due to the low concentration of dicumarol used, which did not affect p21 constitutive levels in control experiments carried out in the absence of the quinone. p21 induction in HT29 cells was also inhibited by DTPA, a metal chelator, and N-acetylcysteine, a potent cellular anti-oxidant, suggesting that HO. may serve as an ultimate mediator for the induction. It may be surmised that the higher efficiency of DZQ in p21 induction may be related to its efficient metabolism by NQOR in HT29 cells and the associated high level of reactive oxygen species. The role of reactive oxygen species in p21 induction was further assessed upon supplementation of cells with H2O2:p21 induction in BE cells was 4-fold higher than that in HT29 cells. These findings suggest that assessment of the role of NQOR and reactive oxygen species in p21 induction requires careful consideration of the cell genotype. PMID- 9343372 TI - Interaction of fibroblast growth factor-1 and related peptides with heparan sulfate and its oligosaccharides. AB - Fibroblast growth factors (FGFs) are a family of angiogenic and mitogenic proteins that promote cell division. The binding of FGFs to the heparan sulfate of cell-surface-bound proteoglycans appears to be critical for their activity. The interaction of fibroblast growth factor-1 (FGF-1 or aFGF) using heparin lyase derived oligosaccharides from heparan sulfate was investigated. FGF-1 was also shown to protect sequences in heparan sulfate from heparin lyase digestion and protected oligosaccharide products of octasaccharide and decasaccharide size were recovered by FGF-1 affinity chromatography, suggesting that the high-affinity binding of heparan sulfate to FGF-1 resides within an octasaccharide sequence. The FGF-1 binding affinity of heparan sulfate is reduced compared to heparin presumably due to the absence of 6-sulfate groups in heparan sulfate. Inspection of the FGF-1 heparan sulfate binding domain shows that the majority of interacting amino acids are contained within a 20-amino-acid sequence that folds back upon itself (because of three turns) forming a triangular shaped cup of positive charge. The importance of FGF-1 binding site topology was investigated using three synthetic peptide mimics of the FGF-1 glycosaminoglycan (GAG) binding site. Heparan sulfate affinity chromatography and isothermal titration calorimetry, used to measure binding thermodynamics, demonstrated that a synthetic peptide analogous to the GAG binding site in FGF-1 bound tightly to heparan sulfate. A peptide containing a D-proline in place of L-proline bound with considerably reduced affinity, presumably due to the altered structure of the second turn in the binding site. A cyclic peptide, expected to be topologically most similar to the triangular GAG binding site in FGF-1, bound with the highest affinity to heparan sulfate. These data suggest the triangular topology of the GAG binding site in FGF is critical for its interaction with heparan sulfate. Analysis of known GAG binding sites in 25 proteins using the Chou-Fasman algorithm show that these sites commonly contain turns. PMID- 9343373 TI - Superoxide-dependent peroxidase activity of H48Q: a superoxide dismutase variant associated with familial amyotrophic lateral sclerosis. AB - Approximately 20% of cases of familial amyotrophic lateral sclerosis are caused by dominant mutations in the Cu,Zn superoxide dismutase. One such mutant, in which histidine #48 has been replaced by glutamine (H48Q), exhibits a novel activity. It can react sequentially with O2- and H2O2 to generate a potent oxidant at its active site, possibly Cu(II)-OH, which then can oxidize urate to the corresponding radical. This O2- -dependent peroxidase activity exerted on a substrate peculiar to motor neurons may be the toxic gain of function which leads to the deleterious consequences of this mutation. G93A, G93R, and E100G were also examined and found not to exert this O2- -dependent peroxidase activity. PMID- 9343374 TI - Distal site and surface mutations of cytochrome P450 1A2 markedly enhance dehalogenation of chlorinated hydrocarbons. AB - Chlorinated compounds such as chlorinated ethylenes and ethanes are serious environmental pollutants. In the present study, we examined whether or not a recombinant strain of Saccharomyces cerevisiae that expresses rat liver cytochrome P450 1A2 (P450 1A2) wild-type and mutant proteins can efficiently catalyze oxidative and reductive dehalogenations of trichloroethylene, pentachloroethane, and hexachloroethane. Mutations at putative heme distal and protein surface sites of P450 1A2 greatly enhanced turnover values toward those substrates under both aerobic and anaerobic conditions. For example, a Thr319Ala mutation at the putative heme distal site enhanced the degradation rate of trichloroethylene and pentachloroethane by 2- and 2.7-fold, respectively, under aerobic conditions. The Thr319Ala mutation also strongly facilitated the reaction with hexachloroethane up to 13- and 4.5-fold under aerobic and anaerobic conditions, respectively. The Thr319Ala mutation increased dechlorinated over protonated product ratios by 3-fold as well when either pentachloroethane or hexachloroethane was used as a substrate. A Lys250Leu mutation on the putative protein surface site enhanced the dehalogenation rate of hexachloroethane up to 4.8-fold under anaerobic conditions. In contrast, a Glu318Ala mutation at the putative distal site markedly decreased the activities with trichloroethylene and pentachloroethane substrates under aerobic conditions. Conserved amino acids Thr319 and Glu318 at the heme distal site have been suggested to be important in the O2 activation during monooxidation reactions of P450s. However, the present study indicates that Thr319 is likely to be an inhibitor of dechlorination of trichloroethylene and penta- and hexachloroethanes. The roles of Thr319, Glu318, and Lys250 in the catalysis with chlorinated hydrocarbons are discussed in association with reaction mechanisms. PMID- 9343375 TI - Human erythrocyte protein L-isoaspartyl methyltransferase: heritability of basal activity and genetic polymorphism for thermal stability. AB - Protein L-isoaspartyl methyltransferase (PIMT) is believed to play an important role in the disposition of age-damaged proteins by catalyzing the repair of abnormal isoaspartyl linkages resulting from the spontaneous deamidation of asparaginyl residues or isomerization of aspartyl residues. As a step toward testing the hypothesis that human disease- or age-related pathology might be associated with a deficiency in PIMT, we investigated basal activity and thermal stability of PIMT in erythrocyte lysates from 299 U.S. family members. Thermal stability was measured because it is a sensitive measure of variation in amino acid sequence. Basal activity was normally distributed with a mean+/-SD of 558+/ 43 units/ml erythrocytes. Statistical analysis of the data revealed that basal PIMT activity exhibited a high degree of heritability. Enzyme thermal stability showed a skewed bimodal frequency distribution, and segregation analysis of family member pedigrees was consistent with Mendelian inheritance of two major alleles. No DNA was available from the family samples, so we tested two additional population samples for a known Ile/Val polymorphism at codon 119 and for PIMT activity and thermal stability, using blood donated by 25 Norwegians and by 20 Koreans. Single-stranded conformational polymorphism analysis using polymerase chain reaction revealed a 100% correlation between thermal stability grouping and this polymorphism. The high thermal stability samples were all homozygous Ile, the low thermal stability samples were all homozygous Val, and the intermediate thermal stability samples were all heterozygous. Furthermore, this polymorphism was responsible, in part, for the variation observed in basal erythrocyte PIMT activity. These results will help provide a foundation for future studies aimed at correlating levels of PIMT activity, or other properties of this enzyme, with human disease. PMID- 9343376 TI - Redox-dependent conformational changes are common structural features of cytochrome c from various species. AB - Discrepant results from X-ray crystallographic and physicochemical studies on the conformations of the two redox states of cytochrome c raise important questions about the nature of redox-dependent conformational changes and whether differences are common structural features of various cytochrome c species. Comparative studies of cytochrome c from 10 species (horse, cow, sheep, pig, dog, rabbit, chicken, pigeon, tuna, and baker's yeast) in aqueous solutions were carried out using Fourier transform infrared (FT-IR) spectroscopy. The second derivative analysis revealed similar conformational changes in all 10 species upon reduction of the heme iron regardless of the differences in the amino acid sequences. The redox-dependent changes involve the amide I regions ascribed to extended beta-structure, beta-turn, and alpha-helix structures. Three species (cow, sheep, and pig) with identical amino acid sequences displayed nearly identical infrared spectra for the oxidized and reduced states, which rules out the possible contribution of experimental error. These results show unequivocally that redox-dependent conformational changes are common structural feature of various cytochrome c species and demonstrate the usefulness of FT-IR spectroscopy as a quick and inexpensive tool in comparative studies of functionally related conformational changes of proteins. PMID- 9343377 TI - Inactivation of the Kluyveromyces lactis H+-ATPase by dicyclohexylcarbodiimide: binding stoichiometry and effect of nucleophiles. AB - Dicyclohexylcarbodiimide (DCCD) inactivated the plasma membrane H+-ATPase (EC 3.6.1.35) from Kluyveromyces lactis, with a second-order rate constant of 420 M( 1) min(-1). The inhibition kinetics was apparently complex, due to degradation of DCCD with time. Neither Mg2+ nor Mg-ADP affected the inactivation of the ATPase by DCCD. In contrast, vanadate, a transition state analog of phosphate, partially protected the enzyme with a Kd of 14 microM, indicating a coupling between the DCCD-reactive site and the vanadate-binding site. The incubation of H+-ATPase with 14C-DCCD showed that the incorporation of 1.2 mol of DCCD/mol ATPase leads to complete inactivation. The hydrophobic carbodiimide reacted with the protonated form of the carboxylic group, which displayed a pKa of 7.4, strongly suggesting that the residue is in the hydrophobic environment of the membrane. Benzylamine increased the rate of inactivation by DCCD. In this case, full inactivation of the enzyme was associated with the incorporation of 2.4 mol of DCCD/mol of enzyme, indicating the opening of new reactive sites, resulting from a conformational change induced by benzylamine. PMID- 9343378 TI - Kinase activity of EnvZ, an osmoregulatory signal transducing protein of Escherichia coli. AB - EnvZ is an inner membrane protein present in Escherichia coli that is important for osmosensing and required for porin gene regulation. EnvZ is phosphorylated by intracellular ATP, and EnvZ-P phosphorylates OmpR, which then binds to the porin promoters to regulate their expression. An overexpressed, truncated form of the enzyme, EnvZ115, was used to characterize the kinase reaction in vitro. Using a filter binding assay, we report the first direct measurements of the kinase activity, including the apparent affinity for ATP of 200 microM. The phosphorylation reaction is dependent on MgCl2, and the phosphoenzyme has the expected stability of a phosphohistidine; i.e., it is stable in base and less stable in acid at room temperature. The addition of OmpR and ATP to solutions containing EnvZ resulted in an OmpR-stimulated, EnvZ-dependent ATPase activity that was not vanadate-sensitive. The in vivo kinase activity of EnvZ and two mutants that were deficient in porin expression were studied using an immune complex kinase reaction. Interestingly, a mutation located in the periplasmic domain of EnvZ exhibited kinase activity that was identical to that of the wild type enzyme, while a mutation located close to the phosphorylation site showed a significant decrease in both kinase and phosphotransferase activities. These data provide support for models of EnvZ consisting of separate sensing and kinase domains. PMID- 9343379 TI - Intracellular localization of Na,K-ATPase alpha2 subunit mutants. AB - The Na,K-ATPase is an essential plasma membrane transporter of mammalian cells composed of two subunits, alpha and beta, of which there are several isoforms. We investigated the effect of a substitution, S364P, on the subcellular localization and enzymatic activity of the wild-type alpha2 and alpha2L111R,N122D (alpha2RD) subunits. The substitutions, L111R and N122D, lower the affinity of the alpha2 subunit for the inhibitor ouabain roughly one thousand-fold (E. A. Jewell and J. B. Lingrel, J. Biol. Chem. 266, 16925-16930, 1991) and were introduced into the alpha2 subunit to distinguish its enzymatic activity from that of the endogenous alpha1 subunit of COS-7 cells. The S364P substitution is located in the ATP binding site, only five residues from the aspartyl residue which is phosphorylated during the catalytic cycle of the Na,K-ATPase. This substitution dramatically decreases the amount of enzymatic activity associated with expression of the alpha2RD subunit. Despite the fact that S364P substitution does not block association of the alpha2RD subunit with the endogenous beta1 subunit, it prevents the alpha2 and alpha2RD subunits from accumulating in the plasma membrane and results in their localization in the endoplasmic reticulum. PMID- 9343380 TI - Stability of Escherichia coli alanyl-tRNA synthetase quaternary structure under increased pressure. PMID- 9343381 TI - Genetic evidence for interaction between Cbp1 and specific nucleotides in the 5' untranslated region of mitochondrial cytochrome b mRNA in Saccharomyces cerevisiae. AB - The cytochrome b (COB) gene is encoded by the mitochondrial genome; however, its expression requires the participation of several nuclearly encoded protein factors. The yeast Cbp1 protein, which is encoded by the nuclear CBP1 gene, is required for the stabilization of COB mRNA. A previous deletion analysis identified an 11-nucleotide-long sequence within the 5' untranslated region of COB mRNA that is important for Cbp1-dependent COB mRNA stability. In the present study, site-directed mutagenesis experiments were carried out to define further the features of this cis element. The CCG sequence within this region was shown to be necessary for stability. A change in residue 533 of Cbp1 from aspartate to tyrosine suppresses the effects of a single-base change in the CCG element. This is strong genetic evidence that the nuclearly encoded Cbp1 protein recognizes and binds directly to the sequence containing CCG and thus protects COB mRNA from degradation. PMID- 9343382 TI - Role for ADA/GCN5 products in antagonizing chromatin-mediated transcriptional repression. AB - The Saccharomyces cerevisiae SWI/SNF complex is a 2-MDa multimeric assembly that facilitates transcriptional enhancement by antagonizing chromatin-mediated transcriptional repression. We show here that mutations in ADA2, ADA3, and GCN5, which are believed to encode subunits of a nuclear histone acetyltransferase complex, cause phenotypes strikingly similar to that of swi/snf mutants. ADA2, ADA3, and GCN5 are required for full expression of all SWI/SNF-dependent genes tested, including HO, SUC2, INO1, and Ty elements. Furthermore, mutations in the SIN1 gene, which encodes a nonhistone chromatin component, or mutations in histone H3 or H4 partially alleviate the transcriptional defects caused by ada/gcn5 or swi/snf mutations. We also find that ada2 swi1, ada3 swi1, and gcn5 swi1 double mutants are inviable and that mutations in SIN1 allow viability of these double mutants. We have partially purified three chromatographically distinct GCN5-dependent acetyltransferase activities, and we show that these enzymes can acetylate both histones and Sin1p. We propose a model in which the ADA/GCN5 and SWI/SNF complexes facilitate activator function by acting in concert to disrupt or modify chromatin structure. PMID- 9343383 TI - Role of UME6 in transcriptional regulation of a DNA repair gene in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae UV radiation and a variety of chemical DNA-damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of these genes is PHR1, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHR1 require an upstream activation sequence, UAS(PHR1), which has homology with DRC elements found upstream of at least 19 other DNA repair and DNA metabolism genes in yeast. Here we report the identification of the UME6 gene of S. cerevisiae as a regulator of UAS(PHR1) activity. Multiple copies of UME6 stimulate expression from UAS(PHR1) and the intact PHR1 gene. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHR1 is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UME6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHR1 mRNA, and increases the UV sensitivity of a rad2 mutant. Despite the fact that UAS(PHR1) does not contain the URS1 sequence, which has been previously implicated in UME6-mediated transcriptional regulation, we find that Ume6p binds to UAS(PHR1) with an affinity and a specificity similar to those seen for a URS1 site. Similar binding is also seen for DRC elements from RAD2, RAD7, and RAD53, suggesting that UME6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. PMID- 9343385 TI - The fission yeast protein p73res2 is an essential component of the mitotic MBF complex and a master regulator of meiosis. AB - Depending on environmental conditions, Schizosaccharomyces pombe can remain in the stationary phase or enter into either premitotic or premeiotic DNA synthesis. This decision point is known as Start. In the mitotic cell cycle, regulation of G1/S-specific gene expression is dependent upon the MBF (Mlu1 binding factor) complex, known to contain p85cdc10 and p72res1. Here we demonstrate that p73res2 controls cell cycle progression via its participation in the MBF complex, interacting directly with both p85cdc10 and p72res1. In contrast, when cells enter into meiosis, the MBF complex is disrupted, and p73res2 shifts its regulatory function towards the transactivation of genes required for meiotic progression. These observations suggest that p73res2 plays a pivotal role at Start and constitutes an example of a transcription factor involved in the control of both mitotic and meiotic progression. PMID- 9343384 TI - Elimination of defective alpha-factor pheromone receptors. AB - This report compares trafficking routes of a plasma membrane protein that was misfolded either during its synthesis or after it had reached the cell surface. A temperature-sensitive mutant form of the yeast alpha-factor pheromone receptor (ste2-3) was found to provide a model substrate for quality control of plasma membrane proteins. We show for the first time that a misfolded membrane protein is recognized at the cell surface and rapidly removed. When the ste2-3 mutant cells were cultured continuously at 34 degrees C, the mutant receptor protein (Ste2-3p) failed to accumulate at the plasma membrane and was degraded with a half-life of 4 min, compared with a half-life of 33 min for wild-type receptor protein (Ste2p). Degradation of both Ste2-3p and Ste2p required the vacuolar proteolytic activities controlled by the PEP4 gene. At 34 degrees C, Ste2-3p comigrated with glycosylated Ste2p on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that Ste2-3p enters the secretory pathway. Degradation of Ste2-3p did not require delivery to the plasma membrane as the sec1 mutation failed to block rapid turnover. Truncation of the C-terminal cytoplasmic domain of the mutant receptors did not permit accumulation at the plasma membrane; thus, the endocytic signals contained in this domain are unnecessary for intracellular retention. In the pep4 mutant, Ste2-3p accumulated as series of high-molecular-weight species, suggesting a potential role for ubiquitin in the elimination process. When ste2-3 mutant cells were cultured continuously at 22 degrees C, Ste2-3p accumulated in the plasma membrane. When the 22 degrees C culture was shifted to 34 degrees C, Ste2-3p was removed from the plasma membrane and degraded by a PEP4-dependent mechanism with a 24-min half life; the wild-type Ste2p displayed a 72-min half-life. Thus, structural defects in Ste2-3p synthesized at 34 degrees C are recognized in transit to the plasma membrane, leading to rapid degradation, and Ste2-3p that is preassembled at the plasma membrane is also removed and degraded following a shift to 34 degrees C. PMID- 9343386 TI - Reciprocal interference between the sequence-specific core and nonspecific C terminal DNA binding domains of p53: implications for regulation. AB - The tumor suppressor p53 has two DNA binding domains: a central sequence-specific domain and a C-terminal sequence-independent domain. Here, we show that binding of large but not small DNAs by the C terminus of p53 negatively regulates sequence-specific DNA binding by the central domain. Four previously described mechanisms for activation of specific DNA binding operate by blocking negative regulation. Deletion of the C terminus of p53 activates specific DNA binding only in the presence of large DNA. Three activator molecules (a small nucleic acid, a monoclonal antibody against the p53 C terminus, and a C-terminal peptide of p53) stimulate sequence-specific DNA binding only in the presence of both large DNA and p53 with an intact C terminus. Our findings argue that interactions of the C terminus of p53 with genomic DNA in vivo would prevent p53 binding to specific promoters and that cellular mechanisms to block C-terminal DNA binding would be required. PMID- 9343387 TI - Neoplastic transformation by truncated alleles of human NOTCH1/TAN1 and NOTCH2. AB - The Notch genes of Drosophila melanogaster and vertebrates encode transmembrane receptors that help determine cell fate during development. Although ligands for Notch proteins have been identified, the signaling cascade downstream of the receptors remains poorly understood. In human acute lymphoblastic T-cell leukemia, a chromosomal translocation damages the NOTCH1 gene. The damage apparently gives rise to a constitutively activated version of NOTCH protein. Here we show that a truncated version of NOTCH1 protein resembling that found in the leukemic cells can transform rat kidney cells in vitro. The transformation required cooperation with the E1A oncogene of adenovirus. The transforming version of NOTCH protein was located in the nucleus. In contrast, neither wild type NOTCH protein nor a form of the truncated protein permanently anchored to the plasma membrane produced transformation in vitro. We conclude that constitutive activation of NOTCH similar to that found in human leukemia can contribute to neoplastic transformation. Transformation may require that the NOTCH protein be translocated to the nucleus. These results sustain a current view of how Notch transduces a signal from the surface of the cell to the nucleus. PMID- 9343388 TI - Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) is required for lipopolysaccharide stimulation of tumor necrosis factor alpha (TNF-alpha) translation: glucocorticoids inhibit TNF-alpha translation by blocking JNK/SAPK. AB - The adverse effects of lipopolysaccharide (LPS) are mediated primarily by tumor necrosis factor alpha (TNF-alpha). TNF-alpha production by LPS-stimulated macrophages is regulated at the levels of both transcription and translation. It has previously been shown that several mitogen-activated protein kinases (MAPKs) are activated in response to LPS. We set out to determine which MAPK signaling pathways are activated in our system and which MAPK pathways are required for TNF alpha gene transcription or TNF-alpha mRNA translation. We confirm activation of the MAPK family members extracellular-signal-regulated kinases 1 and 2 (ERK1 and ERK2), p38, and Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK), as well as activation of the immediate upstream MAPK activators MAPK/ERK kinases 1 and 4 (MEK1 and MEK4). We demonstrate that LPS also activates MEK2, MEK3, and MEK6. Furthermore, we demonstrate that dexamethasone, which inhibits the production of cytokines, including TNF-alpha, significantly inhibits LPS induction of JNK/SAPK activity but not that of p38, ERK1 and ERK2, or MEK3, MEK4, or MEK6. Dexamethasone also blocks the sorbitol but not anisomycin stimulation of JNK/SAPK activity. A kinase-defective mutant of SAPKbeta, SAPKbeta K-A, blocked translation of TNF-alpha, as determined by using a TNF-alpha translational reporting system. Finally, overexpression of wild-type SAPKbeta was able to overcome the dexamethasone-induced block of TNF-alpha translation. These data confirm that three MAPK family members and their upstream activators are stimulated by LPS and demonstrate that JNK/SAPK is required for LPS-induced translation of TNF-alpha mRNA. A novel mechanism by which dexamethasone inhibits translation of TNF-alpha is also revealed. PMID- 9343389 TI - HOY1, a homeo gene required for hyphal formation in Yarrowia lipolytica. AB - The dimorphic fungus Yarrowia lipolytica grows to form hyphae either in rich media or in media with GlcNAc as a carbon source. A visual screening, called FIL (filamentation minus), for Y. lipolytica yeast growth mutants has been developed. The FIL screen was used to identify three Y. lipolytica genes that abolish hypha formation in all media assayed. Y. lipolytica HOY1, a gene whose deletion prevents the yeast-hypha transition both in liquid and solid media, was characterized. HOY1 is predicted to encode a 509-amino-acid protein with a homeodomain homologous to that found in the chicken Hox4.8 gene. Analysis of the protein predicts a nuclear location. These observations suggest that Hoy1p may function as a transcriptional regulatory protein. In disrupted strains, reintroduction of HOY1 restored the capacity for hypha formation. Northern blot hybridization revealed the HOY1 transcript to be approximately 1.6 kb. Expression of this gene was detected when Y. lipolytica grew as a budding yeast, but an increase in its expression was observed by 1 h after cells had been induced to form hyphae. The possible functions of HOY1 in hyphal growth and the uses of the FIL screen to identify morphogenetic regulatory genes from heterologous organisms are discussed. PMID- 9343391 TI - Constitutive expression, not a particular primary sequence, is the important feature of the H3 replacement variant hv2 in Tetrahymena thermophila. AB - Although quantitatively minor replication-independent (replacement) histone variants have been found in a wide variety of organisms, their functions remain unknown. Like the H3.3 replacement variants of vertebrates, hv2, an H3 variant in the ciliated protozoan Tetrahymena thermophila, is synthesized and deposited in nuclei of nongrowing cells. Although hv2 is clearly an H3.3-like replacement variant by its expression, sequence analysis indicates that it evolved independently of the H3.3 variants of multicellular eukaryotes. This suggested that it is the constitutive synthesis, not the particular protein sequence, of these variants that is important in the function of H3 replacement variants. Here, we demonstrate that the gene (HHT3) encoding hv2 or either gene (HHT1 or HHT2) encoding the abundant major H3 can be completely knocked out in Tetrahymena. Surprisingly, when cells lacking hv2 are starved, a major histone H3 mRNA transcribed by the HHT2 gene, which is synthesized little, if at all, in wild-type nongrowing cells, is easily detectable. Both HHT2 and HHT3 knockout strains show no obvious defect during vegetative growth. In addition, a mutant with the double knockout of HHT1 and HHT3 is viable while the HHT2 HHT3 double knockout mutant is not. These results argue strongly that cells require a constitutively expressed H3 gene but that the particular sequence being expressed is not critical. PMID- 9343390 TI - Abortive gap repair: underlying mechanism for Ds element formation. AB - The mechanism by which the maize autonomous Ac transposable element gives rise to nonautonomous Ds elements is largely unknown. Sequence analysis of native maize Ds elements indicates a complex chimeric structure formed through deletions of Ac sequences with or without insertions of Ac-unrelated sequence blocks. These blocks are often flanked by short stretches of reshuffled and duplicated Ac sequences. To better understand the mechanism leading to Ds formation, we designed an assay for detecting alterations in Ac using transgenic tobacco plants carrying a single copy of Ac. We found frequent de novo alterations in Ac which were excision rather than sequence dependent, occurring within Ac but not within an almost identical Ds element and not within a stable transposase-producing gene. The de novo DNA rearrangements consisted of internal deletions with breakpoints usually occurring at short repeats and, in some cases, of duplication of Ac sequences or insertion of Ac-unrelated fragments. The ancient maize Ds elements and the young Ds elements in transgenic tobacco showed similar rearrangements, suggesting that Ac-Ds elements evolve rapidly, more so than stable genes, through deletions, duplications, and reshuffling of their own sequences and through capturing of unrelated sequences. The data presented here suggest that abortive Ac-induced gap repair, through the synthesis-dependent strand-annealing pathway, is the underlying mechanism for Ds element formation. PMID- 9343393 TI - Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct 3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites. AB - Octamer binding and Sox factors are thought to play important roles in development by potentiating the transcriptional activation of specific gene subsets. The proteins within these factor families are related by the presence of highly conserved DNA binding domains, the octamer binding protein POU domain or the Sox factors HMG domain. We have previously shown that fibroblast growth factor 4 (FGF-4) gene expression in embryonal carcinoma cells requires a synergistic interaction between Oct-3 and Sox2 on the FGF-4 enhancer. Sox2 and Oct-3 bind to adjacent sites within this enhancer to form a ternary protein-DNA complex (Oct-3*) whose assembly correlates with enhancer activity. We now demonstrate that increasing the distance between the octamer and Sox binding sites by base pair insertion results in a loss of enhancer function. Significantly, those enhancer "spacing mutants" which failed to activate transcription were also compromised in their ability to form the Oct* complexes even though they could still bind both Sox2 and the octamer binding proteins, suggesting that a direct interaction between Sox2 and Oct-3 is necessary for enhancer function. Consistent with this hypothesis, Oct-3 and Sox2 can participate in a direct protein-protein interaction in vitro in the absence of DNA, and both this interaction and assembly of the ternary Oct* complexes require only the octamer protein POU and Sox2 HMG domains. Assembly of the ternary complex by these two protein domains occurs in a cooperative manner on FGF-4 enhancer DNA, and the loss of this cooperative interaction contributes to the defect in Oct-3* formation observed for the enhancer spacing mutants. These observations indicate that Oct-3* assembly results from protein-protein interactions between the domains of Sox2 and Oct-3 that mediate their binding to DNA, but it also requires a specific arrangement of the binding sites within the FGF-4 enhancer DNA. Thus, these results define one parameter that is fundamental to synergistic activation by Sox2 and Oct-3 and further emphasize the critical role of enhancer DNA sequences in the proper assembly of functional activation complexes. PMID- 9343392 TI - Liver-enriched transcription factors uncoupled from expression of hepatic functions in hepatoma cell lines. AB - Among the liver-enriched transcription factors identified to date, only expression of hepatocyte nuclear factor 4 (HNF4) and hepatocyte nuclear factor 1 (HNF1) is in strict correlation with hepatic differentiation in cultured rat hepatoma cells. Indeed, differentiated hepatoma cells that stably express an extensive set of adult hepatic functions express liver-enriched transcription factors, while dedifferentiated cells that have lost expression of all these hepatic functions no longer express HNF4 and HNF1. We describe a new heritable phenotype, designated as uncoupled, in which there is a spontaneous dissociation between the expression of these transcription factors and that of the hepatic functions. Cells presenting this phenotype, isolated from differentiated hepatoma cells, cease to accumulate all transcripts coding for hepatic functions but nevertheless maintain expression of HNF4 and HNF1. Transitory transfection experiments indicate that these two factors present in these cells have transcriptional activity similar to that of differentiated hepatoma cells. Characterization of the appropriate intertypic cell hybrids demonstrates that this new phenotype is recessive to the dedifferentiated state and fails to be complemented by differentiated cells. These results indicate the existence of mechanisms that inhibit transcription of genes coding for hepatocyte functions in spite of the presence of functional HNF4 and HNF1. Cells of the uncoupled phenotype present certain properties of oval cells described for pathological states of the liver. PMID- 9343394 TI - Transcriptional activation by p53 of the human type IV collagenase (gelatinase A or matrix metalloproteinase 2) promoter. AB - p53, a tumor suppressor and a transcription factor, has been shown to transcriptionally activate the expression of a number of important genes involved in the regulation of cell growth, DNA damage, angiogenesis, and apoptosis. In a computer search for other potential p53 target genes, we identified a perfect p53 binding site in the promoter of the human type IV collagenase (also called 72-kDa gelatinase or matrix metalloproteinase 2 [MMP-2]) gene. This p53 binding site was found to specifically bind to p53 protein in a gel shift assay. Transcription assays with luciferase reporters driven by the promoter or enhancer of the type IV collagenase gene revealed that (i) activation of the promoter activity is p53 binding site dependent in p53-positive cells but not in p53-negative cells and (ii) wild-type p53, but not p53 mutants commonly found in human cancers, transactivates luciferase expression driven by the type IV collagenase promoter as well as by a p53 site-containing enhancer element in the promoter. Significantly, expression of the endogenous type IV collagenase is also under the control of p53. Treatment of U2-OS cells, a wild-type p53-containing osteogenic sarcoma line, with a common p53 inducer, etoposide, induced p53 DNA binding and transactivation activities in a time-dependent manner. Induction of type IV collagenase expression followed the p53 activation pattern. No induction of type IV collagenase expression can be detected under the same experimental conditions in p53-negative Saos-2 cells. All these in vitro and in vivo assays strongly suggest that the type IV collagenase gene is a p53 target gene and that its expression is subject to p53 regulation. Our finding links p53 to a member of the MMP genes, a family of genes implicated in trophoblast implantation, wound healing, angiogenesis, arthritis, and tumor cell invasion. p53 may regulate these processes by upregulating expression of type IV collagenase. PMID- 9343395 TI - A homolog of mammalian, voltage-gated calcium channels mediates yeast pheromone stimulated Ca2+ uptake and exacerbates the cdc1(Ts) growth defect. AB - Previous studies attributed the yeast (Saccharomyces cerevisiae) cdc1(Ts) growth defect to loss of an Mn2+-dependent function. In this report we show that cdc1(Ts) temperature-sensitive growth is also associated with an increase in cytosolic Ca2+. We identified two recessive suppressors of the cdc1(Ts) temperature-sensitive growth which block Ca2+ uptake and accumulation, suggesting that cytosolic Ca2+ exacerbates or is responsible for the cdc1(Ts) growth defect. One of the cdc1(Ts) suppressors is identical to a gene, MID1, recently implicated in mating pheromone-stimulated Ca2+ uptake. The gene (CCH1) corresponding to the second suppressor encodes a protein that bears significant sequence similarity to the pore-forming subunit (alpha1) of plasma membrane, voltage-gated Ca2+ channels from higher eukaryotes. Strains lacking Mid1 or Cch1 protein exhibit a defect in pheromone-induced Ca2+ uptake and consequently lose viability upon mating arrest. The mid1delta and cch1delta mutants also display reduced tolerance to monovalent cations such as Li+, suggesting a role for Ca2+ uptake in the calcineurin dependent ion stress response. Finally, mid1delta cch1delta double mutants are, by both physiological and genetic criteria, identical to single mutants. These and other results suggest Mid1 and Cch1 are components of a yeast Ca2+ channel that may mediate Ca2+ uptake in response to mating pheromone, salt stress, and Mn2+ depletion. PMID- 9343397 TI - Mutations in genes encoding the mitochondrial outer membrane proteins Tom70 and Mdm10 of Podospora anserina modify the spectrum of mitochondrial DNA rearrangements associated with cellular death. AB - Tom70 and Mdm10 are mitochondrial outer membrane proteins. Tom70 is implicated in the import of proteins from the cytosol into the mitochondria in Saccharomyces cerevisiae and Neurospora crassa. Mdm10 is involved in the morphology and distribution of mitochondria in S. cerevisiae. Here we report on the characterization of the genes encoding these proteins in the filamentous fungus Podospora anserina. The two genes were previously genetically identified through a systematic search for nuclear suppressors of a degenerative process displayed by the AS1-4 mutant. The PaTom70 protein shows 80% identity with its N. crassa homolog. The PaMdm10 protein displays 35.9% identity with its S. cerevisiae homolog, and cytological analyses show that the PaMDM10-1 mutant exhibits giant mitochondria, as does the S. cerevisiae mdm10-1 mutant. Mutations in PaTOM70 and PaMDM10 result in the accumulation of specific deleted mitochondrial genomes during the senescence process of the fungus. The phenotypic properties of the single- and double-mutant strains suggest a functional relationship between the Tom70 and Mdm10 proteins. These data emphasize the role of the mitochondrial outer membrane in the stability of the mitochondrial genome in an obligate aerobe, probably through the import process. PMID- 9343396 TI - Retinoid-induced apoptosis and Sp1 cleavage occur independently of transcription and require caspase activation. AB - Vitamin A and its derivatives, the retinoids, are essential regulators of many important biological functions, including cell growth and differentiation, development, homeostasis, and carcinogenesis. Natural retinoids such as all-trans retinoic acid can induce cell differentiation and inhibit growth of certain cancer cells. We recently identified a novel class of synthetic retinoids with strong anti-cancer cell activities in vitro and in vivo which can induce apoptosis in several cancer cell lines. Using an electrophoretic mobility shift assay, we analyzed the DNA binding activity of several transcription factors in T cells treated with apoptotic retinoids. We found that the DNA binding activity of the general transcription factor Sp1 is lost in retinoid-treated T cells undergoing apoptosis. A truncated Sp1 protein is detected by immunoblot analysis, and cytosolic protein extracts prepared from apoptotic cells contain a protease activity which specifically cleaves purified Sp1 in vitro. This proteolysis of Sp1 can be inhibited by N-ethylmaleimide and iodoacetamide, indicating that a cysteine protease mediates cleavage of Sp1. Furthermore, inhibition of Sp1 cleavage by ZVAD-fmk and ZDEVD-fmk suggests that caspases are directly involved in this event. In fact, caspases 2 and 3 are activated in T cells after treatment with apoptotic retinoids. The peptide inhibitors also blocked retinoid-induced apoptosis, as well as processing of caspases and proteolysis of Sp1 and poly(ADP ribose) polymerase in intact cells. Degradation of Sp1 occurs early during apoptosis and is therefore likely to have profound effects on the basal transcription status of the cell. Interestingly, retinoid-induced apoptosis does not require de novo mRNA and protein synthesis, suggesting that a novel mechanism of retinoid signaling is involved, triggering cell death in a transcriptional activation-independent, caspase-dependent manner. PMID- 9343398 TI - Mutations in yeast proliferating cell nuclear antigen define distinct sites for interaction with DNA polymerase delta and DNA polymerase epsilon. AB - The importance of the interdomain connector loop and of the carboxy-terminal domain of Saccharomyces cerevisiae proliferating cell nuclear antigen (PCNA) for functional interaction with DNA polymerases delta (Poldelta) and epsilon (Pol epsilon) was investigated by site-directed mutagenesis. Two alleles, pol30-79 (IL126,128AA) in the interdomain connector loop and pol30-90 (PK252,253AA) near the carboxy terminus, caused growth defects and elevated sensitivity to DNA damaging agents. These two mutants also had elevated rates of spontaneous mutations. The mutator phenotype of pol30-90 was due to partially defective mismatch repair in the mutant. In vitro, the mutant PCNAs showed defects in DNA synthesis. Interestingly, the pol30-79 mutant PCNA (pcna-79) was most defective in replication with Poldelta, whereas pcna-90 was defective in replication with Pol epsilon. Protein-protein interaction studies showed that pcna-79 and pcna-90 failed to interact with Pol delta and Pol epsilon, respectively. In addition, pcna-90 was defective in interaction with the FEN-1 endo-exonuclease (RTH1 product). A loss of interaction between pcna-79 and the smallest subunit of Poldelta, the POL32 gene product, implicates this interaction in the observed defect with the polymerase. Neither PCNA mutant showed a defect in the interaction with replication factor C or in loading by this complex. Processivity of DNA synthesis by the mutant holoenzyme containing pcna-79 was unaffected on poly(dA) x oligo(dT) but was dramatically reduced on a natural template with secondary structure. A stem-loop structure with a 20-bp stem formed a virtually complete block for the holoenzyme containing pcna-79 but posed only a minor pause site for wild-type holoenzyme, indicating a function of the POL32 gene product in allowing replication past structural blocks. PMID- 9343401 TI - Reprogramming of telomerase by expression of mutant telomerase RNA template in human cells leads to altered telomeres that correlate with reduced cell viability. AB - Telomerase synthesizes telomeric DNA by copying the template sequence of its own RNA component. In Tetrahymena thermophila and yeast (G. Yu, J. D. Bradley, L. D. Attardi, and E. H. Blackburn, Nature 344:126-131, 1990; M. McEachern and E. H. Blackburn, Nature 376:403-409, 1995), mutations in the template domain of this RNA result in synthesis of mutant telomeres and in impaired cell growth and survival. We have investigated whether mutant telomerase affects the proliferative potential and viability of immortal human cells. Plasmids encoding mutant or wild-type template RNAs (hTRs) of human telomerase and the neomycin resistance gene were transfected into human cells to generate stable transformants. Expression of mutant hTR resulted in the appearance of mutant telomerase activity and in the synthesis of mutant telomeres. Transformed cells were not visibly affected in their growth and viability when grown as mass populations. However, a reduction in plating efficiency and growth rate and an increase in the number of senescent cells were detected in populations with mutant telomeres by colony-forming assays. These results suggest that the presence of mutant telomerase, even if coexpressed with the wild-type enzyme, can be deleterious to cells, likely as a result of the impaired function of hybrid telomeres. PMID- 9343400 TI - Chromosomal double-strand breaks induce gene conversion at high frequency in mammalian cells. AB - Double-strand breaks (DSBs) stimulate chromosomal and extrachromosomal recombination and gene targeting. Transcription also stimulates spontaneous recombination by an unknown mechanism. We used Saccharomyces cerevisiae I-SceI to stimulate recombination between neo direct repeats in Chinese hamster ovary (CHO) cell chromosomal DNA. One neo allele was controlled by the dexamethasone inducible mouse mammary tumor virus promoter and inactivated by an insertion containing an I-SceI site at which DSBs were introduced in vivo. The other neo allele lacked a promoter but carried 12 phenotypically silent single-base mutations that create restriction sites (restriction fragment length polymorphisms). This system allowed us to generate detailed conversion tract spectra for recipient alleles transcribed at high or low levels. Transient in vivo expression of I-SceI increased homologous recombination 2,000- to 10,000 fold, yielding recombinants at frequencies as high as 1%. Strikingly, 97% of these products arose by gene conversion. Most products had short, bidirectional conversion tracts, and in all cases, donor neo alleles (i.e., those not suffering a DSB) remained unchanged, indicating that conversion was fully nonreciprocal. DSBs in exogenous DNA are usually repaired by end joining requiring little or no homology or by nonconservative homologous recombination (single-strand annealing). In contrast, we show that chromosomal DSBs are efficiently repaired via conservative homologous recombination, principally gene conversion without associated crossing over. For DSB-induced events, similar recombination frequencies and conversion tract spectra were found under conditions of low and high transcription. Thus, transcription does not further stimulate DSB-induced recombination, nor does it appear to affect the mechanism(s) by which DSBs induce gene conversion. PMID- 9343399 TI - V(D)J recombination: in vitro coding joint formation. AB - Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase. PMID- 9343402 TI - The 36-kilodalton embryonic-type cytoplasmic polyadenylation element-binding protein in Xenopus laevis is ElrA, a member of the ELAV family of RNA-binding proteins. AB - The translational activation of several maternal mRNAs in Xenopus laevis is dependent on cytoplasmic poly(A) elongation. Messages harboring the UUUUUAU-type cytoplasmic polyadenylation element (CPE) in their 3' untranslated regions (UTRs) undergo polyadenylation and translation during oocyte maturation. This CPE is bound by the protein CPEB, which is essential for polyadenylation. mRNAs that have the poly(U)12-27 embryonic-type CPE (eCPE) in their 3' UTRs undergo polyadenylation and translation during the early cleavage and blastula stages. A 36-kDa eCPE-binding protein in oocytes and embryos has been identified by UV cross-linking. We now report that this 36-kDa protein is ElrA, a member of the ELAV family of RNA-binding proteins. The proteins are identical in size, antibody directed against ElrA immunoprecipitates the 36-kDa protein, and the two proteins have the same RNA binding specificity in vitro. C12 and activin receptor mRNAs, both of which contain eCPEs, are detected in immunoprecipitated ElrA-mRNP complexes from eggs and embryos. In addition, this in vivo interaction requires the eCPE. Although a number of experiments failed to define a role for ElrA in cytoplasmic polyadenylation, the expression of a dominant negative ElrA protein in embryos results in an exogastrulation phenotype. The possible functions of ElrA in gastrulation are discussed. PMID- 9343403 TI - Transcriptional activation upon pheromone stimulation mediated by a small domain of Saccharomyces cerevisiae Ste12p. AB - In the yeast Saccharomyces cerevisiae, Ste12p induces transcription of pheromone responsive genes by binding to a DNA sequence designated the pheromone response element. We generated a series of hybrid proteins of Ste12p with the DNA-binding and activation domains of the transcriptional activator Gal4p to define a pheromone induction domain of Ste12p sufficient to mediate pheromone-induced transcription by these hybrid proteins. A minimal pheromone induction domain, delineated as residues 301 to 335 of Ste12p, is dependent on the pheromone mitogen-activated protein (MAP) kinase pathway for induction activity. Mutation of the three serine and threonine residues within the minimal pheromone induction domain did not affect transcriptional induction, indicating that the activity of this domain is not directly regulated by MAP kinase phosphorylation. By contrast, mutation of the two tyrosines or their preceding acidic residues led to a high level of transcriptional activity in the absence of pheromone and consequently to the loss of pheromone induction. This constitutively high activity was not affected by mutations in the MAP kinase cascade, suggesting that the function of the pheromone induction domain is normally repressed in the absence of pheromone. By two-hybrid analysis, this minimal domain interacts with two negative regulators, Dig1p and Dig2p (also designated Rst1p and Rst2p), and the interaction is abolished by mutation of the tyrosines. The pheromone induction domain itself has weak and inducible transcriptional activity, and its ability to potentiate transcription depends on the activity of an adjacent activation domain. These results suggest that the pheromone induction domain of Ste12p mediates transcriptional induction via a two-step process: the relief of repression and synergistic transcriptional activation with another activation domain. PMID- 9343404 TI - The Mos pathway regulates cytoplasmic polyadenylation in Xenopus oocytes. AB - Cytoplasmic polyadenylation controls the translation of several maternal mRNAs during Xenopus oocyte maturation and requires two sequences in the 3' untranslated region (UTR), the U-rich cytoplasmic polyadenylation element (CPE), and the hexanucleotide AAUAAA. c-mos mRNA is polyadenylated and translated soon after the induction of maturation, and this protein kinase is necessary for a kinase cascade culminating in cdc2 kinase (MPF) activation. Other mRNAs are polyadenylated later, around the time of cdc2 kinase activation. To determine whether there is a hierarchy in the cytoplasmic polyadenylation of maternal mRNAs, we ablated c-mos mRNA with an antisense oligonucleotide. This prevented histone B4 and cyclin A1 and B1 mRNA polyadenylation, indicating that the polyadenylation of these mRNAs is Mos dependent. To investigate a possible role of cdc2 kinase in this process, cyclin B was injected into oocytes lacking c-mos mRNA. cdc2 kinase was activated, but mitogen-activated protein kinase was not. However, polyadenylation of cyclin B1 and histone B4 mRNA was still observed. This demonstrates that cdc2 kinase can induce cytoplasmic polyadenylation in the absence of Mos. Our data further indicate that although phosphorylation of the CPE binding protein may be involved in the induction of Mos-dependent polyadenylation, it is not required for Mos-independent polyadenylation. We characterized the elements conferring Mos dependence (Mos response elements) in the histone B4 and cyclin B1 mRNAs by mutational analysis. For histone B4 mRNA, the Mos response elements were in the coding region or 5' UTR. For cyclin B1 mRNA, the main Mos response element was a CPE that overlaps with the AAUAAA hexanucleotide. This indicates that the position of the CPE can have a profound influence on the timing of cytoplasmic polyadenylation. PMID- 9343405 TI - Activation of Src family kinases by hepatitis B virus HBx protein and coupled signaling to Ras. AB - The HBx protein of hepatitis B virus (HBV) is a small transcriptional transactivator that is essential for infection by the mammalian hepadnaviruses and is thought to be a cofactor in HBV-mediated liver cancer. HBx stimulates signal transduction pathways by acting in the cytoplasm, which accounts for many but not all of its transcriptional activities. Studies have shown that HBx protein activates Ras and downstream Ras signaling pathways including Raf, mitogen-activated protein (MAP) kinase kinase kinase (MEK), and MAP kinases. In this study, we investigated the mechanism of activation of Ras by HBx because it has been found to be central to the ability of HBx protein to stimulate transcription and to release growth arrest in quiescent cells. In contrast to the transient but strong stimulation of Ras typical of autocrine factors, activation of Ras by HBx protein was found to be constitutive but moderate. HBx induced the association of Ras upstream activating proteins Shc, Grb2, and Sos and stimulated GTP loading onto Ras, but without directly participating in complex formation. Instead, HBx is shown to stimulate Ras-activating proteins by functioning as an intracellular cytoplasmic activator of the Src family of tyrosine kinases, which can signal to Ras. HBx protein stimulated c-Src and Fyn kinases for a prolonged time. Activation of Src is shown to be indispensable for a number of HBx activities, including activation of Ras and the Ras-Raf-MAP kinase pathway and stimulation of transcription mediated by transcription factor AP-1. Importantly, HBx protein expressed in cultured cells during HBV replication is shown to activate the Ras signaling pathway. Mechanisms by which HBx protein might activate Src kinases are discussed. PMID- 9343406 TI - Blockade of T-cell activation by dithiocarbamates involves novel mechanisms of inhibition of nuclear factor of activated T cells. AB - Dithiocarbamates (DTCs) have recently been reported as powerful inhibitors of NF kappaB activation in a number of cell types. Given the role of this transcription factor in the regulation of gene expression in the inflammatory response, NF kappaB inhibitors have been suggested as potential therapeutic drugs for inflammatory diseases. We show here that DTCs inhibited both interleukin 2 (IL-2) synthesis and membrane expression of antigens which are induced during T-cell activation. This inhibition, which occurred with a parallel activation of c-Jun transactivating functions and expression, was reflected by transfection experiments at the IL-2 promoter level, and involved not only the inhibition of NF-kappaB-driven reporter activation but also that of nuclear factor of activated T cells (NFAT). Accordingly, electrophoretic mobility shift assays (EMSAs) indicated that pyrrolidine DTC (PDTC) prevented NF-kappaB, and NFAT DNA-binding activity in T cells stimulated with either phorbol myristate acetate plus ionophore or antibodies against the CD3-T-cell receptor complex and simultaneously activated the binding of AP-1. Furthermore, PDTC differentially targeted both NFATp and NFATc family members, inhibiting the transactivation functions of NFATp and mRNA induction of NFATc. Strikingly, Western blotting and immunocytochemical experiments indicated that PDTC promoted a transient and rapid shuttling of NFATp and NFATc, leading to their accelerated export from the nucleus of activated T cells. We propose that the activation of an NFAT kinase by PDTC could be responsible for the rapid shuttling of the NFAT, therefore transiently converting the sustained transactivation of this transcription factor that occurs during lymphocyte activation, and show that c-Jun NH2-terminal kinase (JNK) can act by directly phosphorylating NFATp. In addition, the combined inhibitory effects on NFAT and NF-KB support a potential use of DTCs as immunosuppressants. PMID- 9343407 TI - AbdB-like Hox proteins stabilize DNA binding by the Meis1 homeodomain proteins. AB - Recent studies show that Hox homeodomain proteins from paralog groups 1 to 10 gain DNA binding specificity and affinity through cooperative binding with the divergent homeodomain protein Pbx1. However, the AbdB-like Hox proteins from paralogs 11, 12, and 13 do not interact with Pbx1a, raising the possibility of different protein partners. The Meis1 homeobox gene has 44% identity to Pbx within the homeodomain and was identified as a common site of viral integration in myeloid leukemias arising in BXH-2 mice. These integrations result in constitutive activation of Meis1. Furthermore, the Hoxa-9 gene is frequently activated by viral integration in the same BXH-2 leukemias, suggesting a biological synergy between these two distinct classes of homeodomain proteins in causing malignant transformation. We now show that the Hoxa-9 protein physically interacts with Meis1 proteins by forming heterodimeric binding complexes on a DNA target containing a Meis1 site (TGACAG) and an AbdB-like Hox site (TTTTACGAC). Hox proteins from the other AbdB-like paralogs, Hoxa-10, Hoxa-11, Hoxd-12, and Hoxb-13, also form DNA binding complexes with Meis1b, while Hox proteins from other paralogs do not appear to interact with Meis1 proteins. DNA binding complexes formed by Meis1 with Hox proteins dissociate much more slowly than DNA complexes with Meis1 alone, suggesting that Hox proteins stabilize the interactions of Meis1 proteins with their DNA targets. PMID- 9343408 TI - Identification of a viral kinase that phosphorylates specific E2Fs and pocket proteins. AB - The transcription factor E2F and its regulation by pRB and related pocket proteins are central to cell cycle control in higher eukaryotes. Much of our knowledge of this regulation has come from studies using immediate-early proteins of DNA tumor viruses. Previously, we reported that the 72-kDa immediate-early region 1 gene product of the human cytomegalovirus, IE72, transactivates the dihydrofolate reductase promoter through the E2F site and that it physically interacts with E2F1 (M. J. Margolis, S. Pajovic, E. L. Wong, M. Wade, R. Jupp, J. A. Nelson, and J. C. Azizkhan, J. Virol. 69:7759-7767, 1995). In this study, we further characterized the mechanism by which IE72 modulates E2F-dependent transcription. In vitro phosphorylation reactions using gel-purified bacterially expressed proteins revealed that IE72 is a kinase that autophosphorylates and phosphorylates E2F1, -2, and -3 (but not E2F4 or -5) and the RB-related pocket proteins p130 and p107 (but not pRB). The region of IE72 spanning amino acids 173 to 197 shows a high level of homology to the ATP binding sites in over 500 kinases. The kinase-negative protein IE72deltaATP, from which this region has been deleted, cannot activate E2F-dependent transcription. The kinase activity of IE72 is also required for its ability to reduce the association of E2F4 with p107 and p130. Taken together, these data suggest that the kinase activity of IE72 is required for E2F-dependent transcriptional activation and that this is likely to result from phosphorylation of specific members of the E2F and pocket protein families by IE72. PMID- 9343409 TI - Characterization of a Holliday junction-resolving enzyme from Schizosaccharomyces pombe. AB - The rearrangement and repair of DNA by homologous recombination involves the creation of Holliday junctions, which are cleaved by a class of junction-specific endonucleases to generate recombinant duplex DNA products. Only two cellular junction-resolving enzymes have been identified to date: RuvC in eubacteria and CCE1 from Saccharomyces cerevisiae mitochondria. We have identified a protein from Schizosaccharomyces pombe which has 28% sequence identity to CCE1. The YDC2 protein has been cloned and overexpressed in Escherichia coli, and the purified recombinant protein has been shown to be a Holliday junction-resolving enzyme. YDC2 has a high degree of specificity for the structure of the four-way junction, to which it binds as a dimer. The enzyme exhibits a sequence specificity for junction cleavage that differs from both CCE1 and RuvC, and it cleaves fixed junctions at the point of strand exchange. The conservation of the mechanism of Holliday junction cleavage between two organisms as diverse as S. cerevisiae and S. pombe suggests that there may be a common pathway for mitochondrial homologous recombination in fungi, plants, protists, and possibly higher eukaryotes. PMID- 9343410 TI - O glycosylation of an Sp1-derived peptide blocks known Sp1 protein interactions. AB - The O-linked N-acetylglucosamine (O-GlcNAc) modification of proteins is dynamic and abundant in the nucleus and cytosol. Several transcription factors, including Sp1, have been shown to contain this modification; however, the functional role of O-GlcNAc in these proteins has not been determined. In this paper we describe the use of the previously characterized glutamine-rich transactivation domain of Sp1 (B-c) as a model to investigate the role of O-GlcNAc in Sp1's transcriptionally relevant protein-to-protein interactions with the TATA-binding protein-associated factor (TAF110) and holo-Sp1. When the model Sp1 peptide was overexpressed in primate cells, this 97-amino-acid domain of Sp1 was found to contain a dominant O-GlcNAc residue at high stoichiometry, which allowed the mapping and mutagenesis of this glycosylation site. In vitro interaction studies between this segment of Sp1 and Drosophila TAF110 or holo-Sp1 indicate that the O GlcNAc modification functions to inhibit the largely hydrophobic interactions between these proteins. In HeLa cells, the mutation at the mapped glycosylation site was permissive for transcriptional activation. We propose the hypothesis that the removal of O-GlcNAc from an interaction domain can be a signal for protein association. O-GlcNAc may thereby prevent untimely and ectopic interactions. PMID- 9343413 TI - Requirements for proteolysis during apoptosis. AB - The key effector proteins of apoptosis are a family of cysteine proteases termed caspases. Following activation of caspases, biochemical events occur that lead to DNA degradation and the characteristic morphological changes associated with apoptosis. Here we show that cytoplasmic extracts activated in vitro by proteinase K were able to cleave the caspase substrate DEVD-7-amino-4 methylcoumarin, while neither proteinase K nor nonactivated extracts were able to do so alone. Caspase-like activity was inhibited by the specific caspase inhibitor DEVD-aldehyde and by the protease inhibitor iodoacetamide, but not by N ethylmaleimide. When added to isolated nuclei, the activated extracts caused internucleosomal DNA degradation and morphological changes typical of apoptosis. As DNA cleavage and morphological changes could be inhibited by N-ethylmaleimide but not by iodoacetamide, we conclude that during apoptosis, caspase activation causes activation of another cytoplasmic enzyme that can be inhibited by N ethylmaleimide. Activity of this enzyme is necessary for activation of endonucleases, DNA cleavage, and changes in nuclear morphology. PMID- 9343412 TI - Xbp1, a stress-induced transcriptional repressor of the Saccharomyces cerevisiae Swi4/Mbp1 family. AB - We have identified Xbp1 (XhoI site-binding protein 1) as a new DNA-binding protein with homology to the DNA-binding domain of the Saccharomyces cerevisiae cell cycle regulating transcription factors Swi4 and Mbp1. The DNA recognition sequence was determined by random oligonucleotide selection and confirmed by gel retardation and footprint analyses. The consensus binding site of Xbp1, GcCTCGA(G/A)G(C/A)g(a/g), is a palindromic sequence, with an XhoI restriction enzyme recognition site at its center. This Xbpl binding site is similar to Swi4/Swi6 and Mbp1/Swi6 binding sites but shows a clear difference from these elements in one of the central core bases. There are binding sites for Xbp1 in the G1 cyclin promoter (CLN1), but they are distinct from the Swi4/Swi6 binding sites in CLN1, and Xbp1 will not bind to Swi4/Swi6 or Mbp1/Swi6 binding sites. The XBP1 promoter contains several stress-regulated elements, and its expression is induced by heat shock, high osmolarity, oxidative stress, DNA damage, and glucose starvation. When fused to the LexA DNA-binding domain, Xbp1 acts as transcriptional repressor, defining it as the first repressor in the Swi4/Mbp1 family and the first potential negative regulator of transcription induced by stress. Overexpression of XBP1 results in a slow-growth phenotype, lengthening of G1, an increase in cell volume, and a repression of G1 cyclin expression. These observations suggest that Xbp1 may contribute to the repression of specific transcripts and cause a transient cell cycle delay under stress conditions. PMID- 9343411 TI - Retinoid-induced chromatin structure alterations in the retinoic acid receptor beta2 promoter. AB - Transcription of the retinoic acid receptor beta2 (RARbeta2) gene is induced by retinoic acid (RA) in mouse P19 embryonal carcinoma (EC) cells. Here we studied RA-induced chromatin structure alterations in the endogenous RARbeta2 promoter and in an integrated, multicopy RARbeta2 promoter in EC cells. RA markedly increased restriction site accessibility within the promoter, including a site near the RA responsive element (RARE) to which the nuclear receptor retinoid X receptor (RXR)-RAR heterodimer binds. These changes coincided with RA-induced alterations in the DNase I hypersensitivity pattern in and around the promoter. These changes became undetectable upon removal of RA, which coincided with the extinction of transcription. Analyses with receptor-selective ligands and an antagonist showed that increase in restriction site accessibility correlates with transcriptional activation, which parallels the RA-induced in vivo footprint of the promoter. Despite these changes, the micrococcal nuclease digestion profile of this promoter was not altered by RA. These results indicate that concurrent with the binding of the RXR-RAR heterodimer to the RARE, the local chromatin structure undergoes dynamic, reversible changes in and around the promoter without globally affecting the nucleosomal organization. PMID- 9343414 TI - STAT3 serine phosphorylation by ERK-dependent and -independent pathways negatively modulates its tyrosine phosphorylation. AB - Recent studies have indicated that serine phosphorylation regulates the activities of STAT1 and STAT3. However, the kinase(s) responsible and the role of serine phosphorylation in STAT function remain unresolved. In the present studies, we examined the growth factor-dependent serine phosphorylation of STAT1 and STAT3. We provide in vitro and in vivo evidence that the ERK family of mitogen-activated protein (MAP) kinases, but not JNK or p38, specifically phosphorylate STAT3 at serine 727 in response to growth factors. Evidence for additional mitogen-regulated serine phosphorylation is also provided. STAT1 is a relatively poor substrate for all MAP kinases tested both in vitro and in vivo. STAT3 serine phosphorylation, not its tyrosine phosphorylation, results in retarded mobility of the STAT3 protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Importantly, serine 727 phosphorylation negatively modulates STAT3 tyrosine phosphorylation, which is required for dimer formation, nuclear translocation, and the DNA binding activity of this transcriptional regulator. Interestingly, the cytokine interleukin-6 also stimulates STAT3 serine phosphorylation, but in contrast to growth factors, this occurs by an ERK independent process. PMID- 9343415 TI - Coordination of the mating and cell integrity mitogen-activated protein kinase pathways in Saccharomyces cerevisiae. AB - Mating pheromone stimulates a mitogen-activated protein (MAP) kinase activation pathway in Saccharomyces cerevisiae that induces cells to differentiate and form projections oriented toward the gradient of pheromone secreted by a mating partner. The polarized growth of mating projections involves new cell wall synthesis, a process that relies on activation of the cell integrity MAP kinase, Mpk1. In this report, we show that Mpk1 activation during pheromone induction requires the transcriptional output of the mating pathway and protein synthesis. Consequently, Mpk1 activation occurs subsequent to the activation of the mating pathway MAP kinase cascade. Additionally, Spa2 and Bni1, a formin family member, are two coil-coil-related proteins that are involved in the timing and other aspects of mating projection formation. Both proteins also affect the timing and extent of Mpk1 activation. This correlation suggests that projection formation comprises part of the pheromone-induced signal that coordinates Mpk1 activation with mating differentiation. Stimulation of Mpk1 activity occurs through the cell integrity phosphorylation cascade and depends on Pkc1 and the redundant MAP/Erk kinases (MEKs), Mkk1 and Mkk2. Surprisingly, Mpk1 activation by pheromone was only partially impaired in cells lacking the MEK kinase Bck1. This Bck1 independent mechanism reveals the existence of an alternative activator of Mkk1/Mkk2 in some strain backgrounds that at least functions under pheromone induced conditions. PMID- 9343417 TI - A complex intronic splicing enhancer from the c-src pre-mRNA activates inclusion of a heterologous exon. AB - The mouse c-src gene contains a short neuron-specific exon, N1. To characterize the sequences that regulate N1 splicing, we used a heterologous gene, derived from the human beta-globin gene, containing a short internal exon that is usually skipped by the splicing machinery. Various fragments from the src gene were inserted into the globin substrate to measure their effects on the splicing of the test exon. These clones were transiently expressed in neuronal and nonneuronal cell lines, and the level of exon inclusion was measured by primer extension. Several sequences from the N1 exon region induced the splicing of the heterologous exon. The most powerful effect was seen with a sequence from the intron downstream of the N1 exon. This sequence acted as a strong splicing enhancer, activating splicing of the test exon when placed in the intron downstream. The enhancer was strongest in neuronal LA-N-5 cells but also activated splicing in nonneuronal HEK293 cells. Deletion and linker scanning mutagenesis indicate that the enhancer is made up of multiple smaller elements that must act in combination. One of these elements was identified as the sequence UGCAUG. Three copies of this element can strongly activate splicing of the test exon in LA-N-5 neuroblastoma cells. These component elements of the src splicing enhancer are also apparently involved in the splicing of other short cassette exons. PMID- 9343416 TI - c-Rel arrests the proliferation of HeLa cells and affects critical regulators of the G1/S-phase transition. AB - A tetracycline-regulated system was used to characterize the effects of c-Rel on cell proliferation. The expression of c-Rel in HeLa cells led to growth arrest at the G1/S-phase transition, which correlated with its nuclear localization and the induction of endogenous IkappaB alpha expression. These changes were accompanied by a decrease in E2F DNA binding and the accumulation of the hypophosphorylated form of Rb. In vitro kinase assays showed a reduction in Cdk2 kinase activity that correlated with elevated levels of p21WAF1 Cdk inhibitor and p53 tumor suppressor protein. While the steady-state levels of WAF1 transcripts were increased, pulse-chase analysis revealed a sharp increase in p53 protein stability. Importantly, the deletion of the C-terminal transactivation domains of c-Rel abolished these effects. Together, these studies demonstrate that c-Rel can affect cell cycle control and suggest the involvement of the p21WAF1 and p53 cell cycle regulators. PMID- 9343418 TI - Proposed mechanism for the stabilization of nuclear receptor DNA binding via protein dimerization. AB - Hepatocyte nuclear factor 4 (HNF-4) defines a new subgroup of nuclear receptors that exist in solution and bind DNA exclusively as homodimers. We recently showed that the putative ligand binding domain (LBD) of HNF-4 is responsible for dimerization in solution and prevents heterodimerization with other receptors. In this report, the role of the LBD in DNA binding by HNF-4 is further investigated by using electrophoretic mobility shift analysis. A comparison of constructs containing either the DNA binding domain (DBD) alone or the DBD plus the LBD of HNF-4 showed that dimerization via the DBD was sufficient to provide nearly the full DNA binding affinity of the full-length HNF-4. In contrast, dimerization via the DBD was not sufficient to produce a stable protein-DNA complex, whereas dimerization via the LBD increased the half-life of the complex by at least 100 fold. Circular permutation analysis showed that full-length HNF-4 bent DNA by approximately 80 degrees while the DBD bent DNA by only 24 degrees. Nonetheless, analysis of other constructs indicated that the increase in stability afforded by the LBD could be explained only partially by an increased ability to bend DNA. Coimmunoprecipitation studies, on the other hand, showed that dimerization via the LBD produced a protein-protein complex that was much more stable than the corresponding protein-DNA complex. These results led us to propose a model in which dimerization via the LBD stabilizes the receptor on DNA by converting an energetically favorable two-step dissociation event into an energetically unfavorable single-step event. Implications of this one-step model for other nuclear receptors are discussed. PMID- 9343419 TI - Amino termini of histones H3 and H4 are required for a1-alpha2 repression in yeast. AB - The Saccharomyces cerevisiae alpha2 repressor controls two classes of cell-type specific genes in yeast through association with different partners. alpha2-Mcm1 complexes repress a cell-specific gene expression in haploid alpha cells and diploid a/alpha cells, while a1-alpha2 complexes repress haploid-specific genes in diploid cells. In both cases, repression is mediated through Ssn6-Tu1 corepressor complexes that are recruited via direct interactions with alpha2. We have previously shown that nucleosomes are positioned adjacent to the alpha2-Mcm1 operator under conditions of repression and that Tupl interacts directly with histones H3 and H4. Here, we examine the role of chromatin in a1-alpha2 repression to determine if chromatin is a general feature of repression by Ssn6 Tup1. We find that mutations in the amino terminus of histone H4 cause a 4- to 11 fold derepression of a reporter gene under a1-alpha2 control, while truncation of the H3 amino terminus has a more modest (3-fold or less) effect. Strikingly, combination of the H3 truncation with an H4 mutation causes a 40-fold decrease in repression, clearly indicating a central role for these histones in a1-alpha2 mediated repression. However, in contrast to the ordered positioning of nucleosomes adjacent to the alpha2-Mcm1 operator, nucleosomes are not positioned adjacent to the a1-alpha2 operator in diploid cells. Our data indicate that chromatin is important to Ssn6-Tup1-mediated repression but that the degrees of chromatin organization directed by these proteins differ at different promoters. PMID- 9343420 TI - The basic domain of myogenic basic helix-loop-helix (bHLH) proteins is the novel target for direct inhibition by another bHLH protein, Twist. AB - In vertebrates, the basic helix-loop-helix (bHLH) protein Twist may be involved in the negative regulation of cellular determination and in the differentiation of several lineages, including myogenesis, osteogenesis, and neurogenesis. Although it has been shown that mouse twist (M-Twist) (i) sequesters E proteins, thus preventing formation of myogenic E protein-MyoD complexes and (ii) inhibits the MEF2 transcription factor, a cofactor of myogenic bHLH proteins, overexpression of E proteins and MEF2 failed to rescue the inhibitory effects of M-Twist on MyoD. We report here that M-Twist physically interacts with the myogenic bHLH proteins in vitro and in vivo and that this interaction is required for the inhibition of MyoD by M-Twist. In contrast to the conventional HLH-HLH domain interaction formed in the MyoD/E12 heterodimer, this novel type of interaction uses the basic domains of the two proteins. While the MyoD HLH domain without the basic domain failed to interact with M-Twist, a MyoD peptide containing only the basic and helix 1 regions was sufficient to interact with M Twist, suggesting that the basic domain contacts M-Twist. The replacement of three arginine residues by alanines in the M-Twist basic domain was sufficient to abolish both the binding and inhibition of MyoD by M-Twist, while the domain retained other M-Twist functions such as heterodimerization with an E protein and inhibition of MEF2 transactivation. These findings demonstrate that M-Twist interacts with MyoD through the basic domains, thereby inhibiting MyoD. PMID- 9343421 TI - The intermembrane space domain of mitochondrial Tom22 functions as a trans binding site for preproteins with N-terminal targeting sequences. AB - Mitochondrial protein import is thought to involve the sequential interaction of preproteins with binding sites on cis and trans sides of the membranes. For translocation across the outer membrane, preproteins first interact with the cytosolic domains of import receptors (cis) and then are translocated through a general import pore, in a process proposed to involve binding to a trans site on the intermembrane space (IMS) side. Controversial results have been reported for the role of the IMS domain of the essential outer membrane protein Tom22 in formation of the trans site. We show with different mutant mitochondria that a lack of the IMS domain only moderately reduces the direct import of preproteins with N-terminal targeting sequences. The dependence of import on the IMS domain of Tom22 is significantly enhanced by removing the cytosolic domains of import receptors or by performing import in two steps, i.e., accumulation of a preprotein at the outer membrane in the absence of a membrane potential (delta psi) and subsequent import after reestablishment of a delta psi. After the removal of cytosolic receptor domains, two-step import of a cleavable preprotein strictly requires the IMS domain. In contrast, preproteins with internal targeting information do not depend on the IMS domain of Tom22. We conclude that the negatively charged IMS domain of Tom22 functions as a trans binding site for preproteins with N-terminal targeting sequences, in agreement with the acid chain hypothesis of mitochondrial protein import. PMID- 9343422 TI - Role of cyclins in neuronal differentiation of immortalized hippocampal cells. AB - The proto-oncogene cyclin D1 and the neuron-specific cyclins p35 and p39 are expressed during brain maturation. To investigate the role of these cyclins in neuronal differentiation, we used a conditionally immortalized rat hippocampal cell line, H19-7, that expresses cyclin-dependent kinases 4 and 5 (cdk4 and -5). Cyclin D1, which activates cdk4 and binds but does not activate cdk5, was increased upon differentiation of the H19-7 cells. However, microinjection of either sense or antisense cyclin D1 cDNA or anti-cyclin D1 antibodies had no effect on morphological differentiation of the cells. On the other hand, neurite outgrowth was stimulated by expression of p35 or p39, both of which activate cdk5. A dominant-negative mutant of cdk5 blocked both p35- and p39-induced neurite extension as well as basic fibroblast growth factor (bFGF)-induced neuronal differentiation. However, of these cyclins, only antisense p39 prevented bFGF-induced neurite outgrowth. These studies indicate that cyclin D1 is neither necessary nor sufficient for morphological differentiation, that p35 is sufficient but not required, and that p39 is both necessary and sufficient for neurite outgrowth in the hippocampal cells. Taken together, these results represent the first demonstration of a specific role for p39 in neuronal differentiation, implicate the cyclin-activated kinase cdk5 in this process, and indicate that p39 is able to mediate neurite outgrowth in the presence or absence of cyclin D1. PMID- 9343423 TI - Inhibition of trans-retinoic acid-resistant human breast cancer cell growth by retinoid X receptor-selective retinoids. AB - All-trans-retinoic acid (trans-RA) and other retinoids exert anticancer effects through two types of retinoid receptors, the RA receptors (RARs) and retinoid X receptors (RXRs). Previous studies demonstrated that the growth-inhibitory effects of trans-RA and related retinoids are impaired in certain estrogen independent breast cancer cell lines due to their lower levels of RAR alpha and RARbeta. In this study, we evaluated several synthetic retinoids for their ability to induce growth inhibition and apoptosis in both trans-RA-sensitive and trans-RA-resistant breast cancer cell lines. Our results demonstrate that RXR selective retinoids, particularly in combination with RAR-selective retinoids, could significantly induce RARbeta and inhibit the growth and induce the apoptosis of trans-RA-resistant, RAR alpha-deficient MDA-MB-231 cells but had low activity against trans-RA-sensitive ZR-75-1 cells that express high levels of RAR alpha. Using gel retardation and transient transfection assays, we found that the effects of RXR-selective retinoids on MDA-MB-231 cells were most likely mediated by RXR-nur77 heterodimers that bound to the RA response element in the RARbeta promoter and activated the RARbeta promoter in response to RXR-selective retinoids. In contrast, growth inhibition by RAR-selective retinoids in trans-RA sensitive, RAR alpha-expressing cells most probably occurred through RXR-RAR alpha heterodimers that also bound to and activated the RARbeta promoter. In MDA MB-231 clones stably expressing RAR alpha, both RARbeta induction and growth inhibition by RXR-selective retinoids were suppressed, while the effects of RAR selective retinoids were enhanced. Together, our results demonstrate that activation of RXR can inhibit the growth of trans-RA-resistant MDA-MB-231 breast cancer cells and suggest that low cellular RAR alpha may regulate the signaling switch from RAR-mediated to RXR-mediated growth inhibition in breast cancer cells. PMID- 9343425 TI - Stat1 serine phosphorylation occurs independently of tyrosine phosphorylation and requires an activated Jak2 kinase. AB - Gamma interferon (IFN-gamma) induces both tyrosine and serine phosphorylation of Stat1. Stat1 serine phosphorylation is required for maximal transcriptional activity of Stat1. In this report, we present evidence that Stat1 tyrosine phosphorylation is not a prerequisite for Stat1 serine phosphorylation, although an active Jak2 kinase is required for both phosphorylation events. Stat1 serine phosphorylation occurs with a more delayed time course than tyrosine phosphorylation. The occurrence of serine phosphorylation without tyrosine phosphorylation suggests that serine phosphorylation takes place in the cytoplasm. Experiments performed with cells expressing either dominant-negative or constitutively active Ras protein indicated that the Ras-mitogen-activated protein kinase pathway is probably not involved in IFN-gamma-induced Stat1 serine phosphorylation. Finally, a kinase capable of correct Stat1 serine phosphorylation was detected in partially purified cytoplasmic extracts from both IFN-gamma-treated and untreated cells. PMID- 9343424 TI - Interaction and functional collaboration of p300 and C/EBPbeta. AB - Transcriptional coactivators such as p300 and CREB-binding protein (CBP) function as important elements in the transcription factor network, linking individual transactivators via protein-protein interactions to the basal transcriptional machinery. We have investigated whether p300 plays a role in transactivation mediated by C/EBPbeta, a conserved member of the C/EBP family. We show that C/EBPbeta-dependent transactivation is strongly inhibited by adenovirus E1A but not by E1A mutants defective in p300 binding. Ectopic expression of p300 reverses the E1A-dependent inhibition and increases the transactivation potential of C/EBPbeta. Furthermore, we show that C/EBPbeta and p300 interact with each other and demonstrate that the sequences responsible for interaction map to the E1A binding region of p300 and the amino terminus of C/EBPbeta. Finally, we show that the minimal C/EBPbeta binding site of p300 acts as a dominant-negative inhibitor of C/EBPbeta. These observations identify p300 as a bona fide coactivator for C/EBPbeta. C/EBPbeta is highly expressed in the myelomonocytic lineage of the hematopoietic system and cooperates with Myb to activate mim-1, a gene specifically expressed during myelomonocytic differentiation. Recent evidence has shown that Myb recruits CBP (and presumably p300) as a coactivator and, in contrast to C/EBPbeta, interacts with the CREB binding site of p300-CBP. We show that p300 not only stimulates the activity of Myb and C/EBPbeta individually but also increases the synergy between them. Thus, our results reveal a novel function of p300: in addition to linking specific transcription factors to the basal transcriptional machinery, p300 also mediates the cooperation between transactivators interacting with different domains of p300. PMID- 9343426 TI - Requirement for transcription factor IIA (TFIIA)-TFIID recruitment by an activator depends on promoter structure and template competition. AB - Different mechanisms of transcriptional activation may be required for distinct classes of promoters and cellular conditions. The Epstein-Barr virus (EBV) encoded transcriptional activator Zta recruits the general transcription factors IID (TFIID) and IIA (TFIIA) to promoter DNA and induces a TATA box-binding protein (TBP)-associated factor-dependent footprint downstream of the transcriptional initiation site. In this study, we investigated the functional significance of TFIID-TFIIA (D-A complex) recruitment by Zta. Alanine substitution mutations in the Zta activation domain which eliminate the ability of Zta to stimulate the D-A complex were examined. These Zta mutants were defective in the ability to activate transcription from an EBV-derived promoter (BHLF1) but activated a highly responsive synthetic promoter (Z7E4T). Both the number of activator binding sites and the core promoter region contribute to the requirement for D-A complex recruitment. These functionally distinct core promoters had significant differences in affinity for TBP and TFIID binding. The D-A complex-recruiting activity of Zta was found to be important for promoter selection in the presence of a competitor template. Conditions which limit TFIID binding to the TATA element or compromise the ability of TFIIA to bind TBP required activator stimulation of the D-A complex. These results indicate that D A complex recruitment is one of at least two activation pathways utilized by Zta and is the essential pathway for a subset of promoters and conditions which limit TFIID binding to the TATA element. PMID- 9343427 TI - Identification of SH2-Bbeta as a substrate of the tyrosine kinase JAK2 involved in growth hormone signaling. AB - Activation of the tyrosine kinase JAK2 is an essential step in cellular signaling by growth hormone (GH) and multiple other hormones and cytokines. Murine JAK2 has a total of 49 tyrosines which, if phosphorylated, could serve as docking sites for Src homology 2 (SH2) or phosphotyrosine binding domain-containing signaling molecules. Using a yeast two-hybrid screen of a rat adipocyte cDNA library, we identified a splicing variant of the SH2 domain-containing protein SH2-B, designated SH2-Bbeta, as a JAK2-interacting protein. The carboxyl terminus of SH2 Bbeta (SH2-Bbetac), which contains the SH2 domain, specifically interacts with kinase-active, tyrosyl-phosphorylated JAK2 but not kinase-inactive, unphosphorylated JAK2 in the yeast two-hybrid system. In COS cells coexpressing SH2-Bbeta or SH2-Bbetac and murine JAK2, both SH2-Bbetac and SH2-Bbeta coimmunoprecipitate to a significantly greater extent with wild-type, tyrosyl phosphorylated JAK2 than with kinase-inactive, unphosphorylated JAK2. SH2-Bbetac also binds to immunoprecipitated wild-type but not kinase-inactive JAK2 in a far Western blot. In 3T3-F442A cells, GH stimulates the interaction of SH2-Bbeta with tyrosyl-phosphorylated JAK2 both in vitro, as assessed by binding of JAK2 in cell lysates to glutathione S-transferase (GST)-SH2-Bbetac or GST-SH2-Bbeta fusion proteins, and in vivo, as assessed by coimmunoprecipitation of JAK2 with SH2 Bbeta. GH promoted a transient and dose-dependent tyrosyl phosphorylation of SH2 Bbeta in 3T3-F442A cells, further suggesting the involvement of SH2-Bbeta in GH signaling. Consistent with SH2-Bbeta being a substrate of JAK2, SH2-Bbetac is tyrosyl phosphorylated when coexpressed with wild-type but not kinase-inactive JAK2 in both yeast and COS cells. SH2-Bbeta was also tyrosyl phosphorylated in response to gamma interferon, a cytokine that activates JAK2 and JAK1. These data suggest that GH-induced activation and phosphorylation of JAK2 recruits SH2-Bbeta and its associated signaling molecules into a GHR-JAK2 complex, thereby initiating some as yet unidentified signal transduction pathways. These pathways are likely to be shared by other cytokines that activate JAK2. PMID- 9343428 TI - Processing of targeted psoralen cross-links in Xenopus oocytes. AB - Psoralen cross-links have been shown to be both mutagenic and recombinagenic in bacterial, yeast, and mammalian cells. Double-strand breaks (DSBs) have been implicated as intermediates in the removal of psoralen cross-links. Recent work has suggested that site-specific mutagenesis and recombination might be achieved through the use of targeted psoralen adducts. The fate of plasmids containing psoralen adducts was evaluated in Xenopus oocytes, an experimental system that has well-characterized recombination capabilities and advantages in the analysis of intermediates in DNA metabolism. Psoralen adducts were delivered to a specific site by a triplex-forming oligonucleotide. These lesions are clearly recognized and processed in oocytes, since mutagenesis was observed at the target site. The spectrum of induced mutations was compared with that found in similar studies in mammalian cells. Plasmids carrying multiple random adducts were preferentially degraded, perhaps due to the introduction of DSBs. However, when DNAs carrying site-specific adducts were examined, no plasmid loss was observed and removal of cross-links was found to be very slow. Sensitive assays for DSB-dependent homologous recombination were performed with substrates with one or two cross link sites. No adduct-stimulated recombination was observed with a single lesion, and only very low levels were observed with paired lesions, even when a large proportion of the cross-links was removed by the oocytes. We conclude that DSBs or other recombinagenic structures are not efficiently formed at psoralen adducts in Xenopus oocytes. While psoralen is not a promising reagent for stimulating site-specific recombination, it is effective in inducing targeted mutations. PMID- 9343429 TI - Protein and DNA contact surfaces that mediate the selective action of the Phox1 homeodomain at the c-fos serum response element. AB - The human homeodomain protein Phox1 can impart serum-responsive transcriptional activity to the c-fos serum response element (SRE) by interacting with serum response factor (SRF). This activity is shared with other Paired class homeodomains but not with more distantly related homeodomains. To understand the mechanism of action of Phox1 at the SRE and the basis for the selective activity of Paired class homeodomains in this context, we performed a detailed mutagenesis of the Phox1 homeodomain. We found that amino acid residues that contact the major groove of the DNA are required for SRE activation in vivo, suggesting an in vivo requirement for major-groove DNA contact by the homeodomain. In contrast, substitution of a lysine residue in the N-terminal arm of the Phox1 homeodomain appeared to abolish DNA binding without affecting activity in vivo. Certain substitutions on the exposed surfaces of helices 1 and 2, not required for DNA binding, abolished activity in vivo, suggesting that these surfaces contact an accessory protein(s) required for this activity. We also found that transfer of a single amino acid residue from the surface of Phox1 helix 1 to the corresponding position in the distantly related Deformed (Dfd) homeodomain imparts to Dfd the ability to activate the SRE in vivo. We propose that Phox1 interacts with one or more factors at the SRE, in addition to SRF, and that the specificity of this interaction is determined by residues on the surfaces of helices 1 and 2. PMID- 9343430 TI - Evolutionary conservation and predicted structure of the Drosophila extra sex combs repressor protein. AB - The Drosophila extra sex combs (esc) protein, a member of the Polycomb group (PcG), is a transcriptional repressor of homeotic genes. Genetic studies have shown that esc protein is required in early embryos at about the time that other PcG proteins become engaged in homeotic gene repression. The esc protein consists primarily of multiple copies of the WD repeat, a motif that has been implicated in protein-protein interaction. To further investigate the domain organization of esc protein, we have isolated and characterized esc homologs from divergent insect species. We report that esc protein is highly conserved in housefly (72% identical to Drosophila esc), butterfly (55% identical), and grasshopper (56% identical). We show that the butterfly homolog provides esc function in Drosophila, indicating that the sequence similarities reflect functional conservation. Homology modeling using the crystal structure of another WD repeat protein, the G-protein beta-subunit, predicts that esc protein adopts a beta propeller structure. The sequence comparisons and modeling suggest that there are seven WD repeats in esc protein which together form a seven-bladed beta propeller. We locate the conserved regions in esc protein with respect to this predicted structure. Site-directed mutagenesis of specific loops, predicted to extend from the propeller surface, identifies conserved parts of esc protein required for function in vivo. We suggest that these regions might mediate physical interaction with esc partner proteins. PMID- 9343431 TI - NeuroD1/beta2 contributes to cell-specific transcription of the proopiomelanocortin gene. AB - NeuroD1/beta2 is a basic helix-loop-helix (bHLH) factor expressed in the endocrine cells of the pancreas and in a subset of neurons as they undergo terminal differentiation. We now show that NeuroD1 is expressed in corticotroph cells of the pituitary gland and that it is involved in cell-specific transcription of the proopiomelanocortin (POMC) gene. It was previously shown that corticotroph-specific POMC transcription depends in part on the action of cell-restricted bHLH factors that were characterized as the CUTE (corticotroph upstream transcription element) (M. Therrien and J. Drouin, Mol. Cell. Biol. 13:2342-2353, 1993) complexes. We now demonstrate that these complexes contain NeuroD1 in association with various ubiquitous bHLH dimerization partners. The NeuroD1-containing heterodimers specifically recognize and activate transcription from the POMC promoter E box that confers transcriptional specificity. Interestingly, the NeuroD1 heterodimers activate transcription in synergy with Ptx1, a Bicoid-related homeodomain protein, which also contributes to corticotroph specificity of POMC transcription. In the adult pituitary gland, NeuroD1 transcripts are detected in POMC-expressing corticotroph cells. Taken together with the restricted pattern of Ptx1 expression, these results suggest that these two factors establish the basis of a combinatorial code for the program of corticotroph-specific gene expression. PMID- 9343432 TI - A domain shared by the Polycomb group proteins Scm and ph mediates heterotypic and homotypic interactions. AB - The Sex comb on midleg (Scm) and polyhomeotic (ph) proteins are members of the Polycomb group (PcG) of transcriptional repressors. PcG proteins maintain differential patterns of homeotic gene expression during development in Drosophila flies. The Scm and ph proteins share a homology domain with 38% identity over a length of 65 amino acids, termed the SPM domain, that is located at their respective C termini. Using the yeast two-hybrid system and in vitro protein-binding assays, we show that the SPM domain mediates direct interaction between Scm and ph. Binding studies with isolated SPM domains from Scm and ph show that the domain is sufficient for these protein interactions. These studies also show that the Scm-ph and Scm-Scm domain interactions are much stronger than the ph-ph domain interaction, indicating that the isolated domain has intrinsic binding specificity determinants. Analysis of site-directed point mutations identifies residues that are important for SPM domain function. These binding properties, predicted alpha-helical secondary structure, and conservation of hydrophobic residues prompt comparisons of the SPM domain to the helix-loop-helix and leucine zipper domains used for homotypic and heterotypic protein interactions in other transcriptional regulators. In addition to in vitro studies, we show colocalization of the Scm and ph proteins at polytene chromosome sites in vivo. We discuss the possible roles of the SPM domain in the assembly or function of molecular complexes of PcG proteins. PMID- 9343433 TI - The ubiquitin-conjugating enzyme Rad6 (Ubc2) is required for silencing in Saccharomyces cerevisiae. AB - It has been previously shown that genes transcribed by RNA polymerase II (RNAP II) are subject to position effect variegation when located near yeast telomeres. This telomere position effect requires a number of gene products that are also required for silencing at the HML and HMR loci. Here, we show that a null mutation of the DNA repair gene RAD6 reduces silencing of the HM loci and lowers the mating efficiency of MATa strains. Likewise, rad6-delta reduces silencing of the telomere-located RNAP II-transcribed genes URA3 and ADE2. We also show that the RNAP III-transcribed tyrosyl tRNA gene, SUP4-o, is subject to position effect variegation when located near a telomere and that this silencing requires the RAD6 and SIR genes. Neither of the two known Rad6 binding factors, Rad18 and Ubr1, is required for telomeric silencing. Since Ubrl is the recognition component of the N-end rule-dependent protein degradation pathway, this suggests that N-end rule-dependent protein degradation is not involved in telomeric silencing. Telomeric silencing requires the amino terminus of Rad6. Two rad6 point mutations, rad6(C88A) and rad6(C88S), which are defective in ubiquitin conjugating activity fail to complement the silencing defect, indicating that the ubiquitin-conjugating activity of RAD6 is essential for full telomeric silencing. PMID- 9343434 TI - gadd153/Chop10, a potential target gene of the transcriptional repressor ATF3. AB - Recently, we demonstrated that the function of ATF3, a stress-inducible transcriptional repressor, is negatively regulated by a bZip protein, gadd153/Chop10. In this report, we present evidence that ATF3 can repress the expression of its own inhibitor, gadd153/Chop10. First, ATF3 represses a chloramphenicol acetyltransferase reporter gene driven by the gadd153/Chop10 promoter when assayed by a transfection assay in vivo and a transcription assay in vitro. Second, the gadd153/Chop10 promoter contains two functionally important binding sites for ATF3: an AP-1 site and a C/EBP-ATF composite site, a previously unidentified binding site for ATF3. The absence of either site reduces the ability of ATF3 to repress the promoter. Third, overexpression of ATF3 by transient transfection results in a reduction of the endogenous gadd153/Chop10 mRNA level. Fourth, as described previously, ATF3 is induced in the liver upon CCl4 treatment. Intriguingly, we show in this report that gadd153/Chop10 mRNA is not present in areas where ATF3 is induced. Taken together, these results strongly suggest that ATF3 represses the expression of gadd153/Chop10. The mutual negative regulation between ATF3 and gadd153/Chop10 is discussed. PMID- 9343436 TI - Activation of cyclic AMP-dependent kinase is required but may not be sufficient to mimic cyclic AMP-dependent DNA synthesis and thyroglobulin expression in dog thyroid cells. AB - Thyrotropin (TSH), via a cyclic AMP (cAMP)-dependent pathway, induces cytoplasmic retractions, proliferation, and differentiation expression in dog thyroid cells. The role of cAMP-dependent protein kinase (PKA) in the induction of these events was assessed by microinjection into living cells. Microinjection of the heat stable inhibitor of PKA (PKI) inhibited the effects of TSH, demonstrating that activation of PKA was required in this process. Overexpression of the catalytic (C) subunit of PKA brought about by microinjection of the expression plasmid pC alpha ev or of purified C subunit itself was sufficient to mimic the cAMP dependent cytoplasmic changes and thyroperoxidase mRNA expression but not to induce DNA synthesis and thyroglobulin (Tg) expression. The cAMP-dependent morphological effect was not observed when C subunit was coinjected with the regulatory subunit (RI or RII subunit) of PKA. To mimic the cAMP-induced PKA dissociation into free C and R subunits, the C subunit was coinjected with the regulation-deficient truncated RI subunit (RIdelta1-95) or with wild-type RI or native RII subunits, followed by incubation with TSH at a concentration too low to stimulate the cAMP-dependent events by itself. Although the cAMP-dependent morphology changes were still observed, neither DNA synthesis nor Tg expression was stimulated in these cells. Taken together, these data suggest that in addition to PKA activation, another cAMP-dependent mechanism could exist and play an important role in the transduction of the cAMP signal in thyroid cells. PMID- 9343435 TI - Specific DNA binding of Stat5, but not of glucocorticoid receptor, is required for their functional cooperation in the regulation of gene transcription. AB - Prolactin and glucocorticoid hormone are signals which regulate the transcription of milk protein genes in mammary epithelial cells. We have investigated the molecular mechanisms by which these hormones cooperate in the induction of transcription. Both hormones activate latent transcription factors in the cytoplasm of mammary epithelial cells. Prolactin exerts its effect through binding to the extracellular domain of the prolactin receptor and through receptor dimerization. This leads to the activation of a protein tyrosine kinase (Jak2), which is noncovalently associated with the cytoplasmic domain of the prolactin receptor. Jak2 phosphorylates the signal transducer and transcription activator (Stat5) which causes its dimerization and nuclear translocation where Stat5 specifically binds to sequence elements in the promoter regions of milk protein genes. In comparison, the glucocorticoid receptor is activated by a lipophilic steroid ligand in the cytoplasm which causes allosteric changes in the molecule, dimerization, and nuclear localization. It has been demonstrated that Stat5 and the glucocorticoid receptor form a molecular complex which cooperates in the induction of transcription of the beta-casein gene. We have defined the DNA sequence requirements for this cooperative mechanism and have delimited the functional domains in Stat5 and the glucocorticoid receptor that are necessary for the functional interaction. We find that the Stat5 response element (Stat5RE) within the beta-casein gene promoter is sufficient to elicit the cooperative action of Stat5 and the glucocorticoid receptor on transcription. Activation of Stat5 through phosphorylation of tyrosine 694 is an absolute prerequisite for transcription. Deletion of the transactivation domain of Stat5 results in a molecule which cannot mediate transactivation by itself but can still cooperate with the glucocorticoid receptor. Mutated variants of the glucocorticoid receptor with a nonfunctional DNA binding domain or a DNA binding domain contributed by the estrogen receptor are still able to cooperate with Stat5 in transcriptional induction. Deletion of the ligand binding domain of the glucocorticoid receptor does not impede cooperation with Stat5, whereas deletion of the AF-1 transactivation domain does prevent cooperation. Our results indicate that the glucocorticoid receptor acts as a ligand-dependent coactivator of Stat5 independently of its DNA binding function. PMID- 9343437 TI - Tissue-specific splicing and functions of the Drosophila transcription factor Grainyhead. AB - Grainyhead belongs to a recently identified group of transcription factors which share a 250-amino-acid domain required for binding to DNA and a carboxy-terminal dimerization domain. The activities of Grainyhead and other members of the family appear to be modulated so that they can participate in different developmental processes. We have examined the structure and function of mRNAs from the Drosophila grainyhead gene and demonstrated that alternate splicing is responsible for generating a neuroblast-specific isoform of the protein. A mutation which abolishes this isoform results in pupal and adult lethality. Reporter genes containing different Grainyhead binding sites exhibit tissue specific patterns of expression that correlate with the Grainyhead isoforms, suggesting that the alternate splicing serves to alter the repertoire of target genes controlled in the neuroblasts. PMID- 9343438 TI - A functional role for death proteases in s-Myc- and c-Myc-mediated apoptosis. AB - Upon activation, cell surface death receptors, Fas/APO-1/CD95 and tumor necrosis factor receptor-1 (TNFR-1), are attached to cytosolic adaptor proteins, which in turn recruit caspase-8 (MACH/FLICE/Mch5) to activate the interleukin-1 beta converting enzyme (ICE)/CED-3 family protease (caspase) cascade. However, it remains unknown whether these apoptotic proteases are generally involved in apoptosis triggered by other stimuli such as Myc and p53. In this study, we provide lines of evidence that a death protease cascade consisting of caspases and serine proteases plays an essential role in Myc-mediated apoptosis. When Rat 1 fibroblasts stably expressing either s-Myc or c-Myc were induced to undergo apoptosis by serum deprivation, a caspase-3 (CPP32)-like protease activity that cleaves a specific peptide substrate, Ac-DEVD-MCA, appeared in the cell lysates. Induction of s-Myc- and c-Myc-mediated apoptotic cell death was effectively prevented by caspase inhibitors such as Z-Asp-CH2-DCB and Ac-DEVD-CHO. Furthermore, exposing the cells to a serine protease inhibitor, 4-(2 aminoethyl)benzenesulfonyl fluoride (AEBSF), also significantly inhibited s-Myc- and c-Myc-mediated apoptosis and the appearance of the caspase-3-like protease activity in vivo. However, AEBSF did not directly inhibit caspase-3-like protease activity in the apoptotic cell lysates in vitro. Together, these results indicate that caspase-3-like proteases play a critical role in both s-Myc- and c-Myc mediated apoptosis and that caspase-3-like proteases function downstream of the AEBSF-sensitive step in the signaling pathway of Myc-mediated apoptosis. PMID- 9343439 TI - Synergistic activation of NF-kappaB by tumor necrosis factor alpha and gamma interferon via enhanced I kappaB alpha degradation and de novo I kappaBbeta degradation. AB - Tumor necrosis factor alpha (TNF-alpha) and gamma interferon (IFN-gamma) are required for an effective immune response to bacterial infection and these cytokines synergize in a variety of biological responses, including the induction of cytokine, cell adhesion, and inducible nitrous oxide synthase gene expression. Typically, the synergistic effect on gene expression is due to the independent activation of nuclear factor kappaB (NF-kappaB) by TNF-alpha and of signal transducers and activators of transcription or IFN-regulatory factor 1 by IFNs, allowing these transcription factors to bind their unique promoter sites. However, since activation of NF-kappaB by TNF-alpha is often transient and would not activate long-term kappaB-dependent transcription effectively, we explored the effects of IFN-gamma on TNF-alpha-induced NF-kappaB activity. IFN-gamma, which typically does not activate NF-kappaB, synergistically enhanced TNF-alpha induced NF-kappaB nuclear translocation via a mechanism that involves the induced degradation of I kappaBbeta and that apparently requires tyrosine kinase activity in preneuronal cells but not in endothelial cells. Correspondingly, cotreatment of cells with TNF-alpha and IFN-gamma leads to persistent activation of NF-kappaB and to potent activation of kappaB-dependent gene expression, which may explain, at least in part, the synergy observed between these cytokines, as well as their involvement in the generation of an effective immune response. PMID- 9343444 TI - FDA sets geriatric drug use labeling deadlines. PMID- 9343440 TI - A role for c-myc in chemically induced renal-cell death. AB - A variety of genes, including c-myc, are activated by chemical toxicants in vivo and in vitro. Although enforced c-myc expression induces apoptosis after withdrawing survival factors, it is not clear if activation of the endogenous c myc gene is an apoptotic signal after toxicant exposure. The renal tubular epithelium is a target for many toxicants. c-myc expression is activated by tubular damage. In quiescent LLC-PK1 renal epithelial cells, c-myc but not max or mad mRNA is induced by the nephrotoxicant S-(1,2-dichlorovinyl)-L-cysteine (DCVC). The kinetics of DCVC-induced c-myc expression and apoptosis suggested an association between cell death and prolonged activation of c-myc expression after toxicant exposure. Accordingly, prolonged activation of an estrogen receptor-Myc fusion construct, but not a construct in which a c-Myc transactivation domain had been deleted, was sufficient to induce apoptosis in LLC-PK1 cells. Moreover, under conditions in which necrosis was the predominant cell death pathway caused by DCVC in parental cells, overexpressing c-myc biased the cell death pathway toward apoptosis. DCVC also induced ornithine decarboxylase (odc) mRNA and activated the odc promoter. Activation of the odc promoter by DCVC required consensus c-Myc-Max binding sites in odc intron 1. Inhibiting ODC activity with alpha-difluoromethylornithine delayed DCVC-induced cell death. Therefore, odc is a target gene in the DCVC apoptotic pathway involving c-myc activation and contributes to apoptosis. Finally, a structurally related cytotoxic but nongenotoxic analog of DCVC did not induce c-myc and did not activate the odc promoter or induce apoptosis. The data support the hypothesis that activation of apoptotic cell death in quiescent renal epithelial cells involves induction of c myc. This is the first study to demonstrate that c-myc induction by a specific nephrotoxicant leads to gene activation and cell death. PMID- 9343443 TI - Federal program nourishes poor elderly. PMID- 9343442 TI - Vintage care. Geriatrics resources on the Net. PMID- 9343445 TI - 'Old and gray and full of sleep'? Not always. PMID- 9343441 TI - Two pathways for removal of nonhomologous DNA ends during double-strand break repair in Saccharomyces cerevisiae. AB - During repair of a double-strand break (DSB) by gene conversion, one or both 3' ends of the DSB invade a homologous donor sequence and initiate new DNA synthesis. The use of the invading DNA strand as a primer for new DNA synthesis requires that any nonhomologous bases at the 3' end be removed. We have previously shown that removal of a 3' nonhomologous tail in Saccharomyces cerevisiae depends on the nucleotide excision repair endonuclease Rad1/Rad10, and also on the mismatch repair proteins Msh2 and Msh3. We now report that these four proteins are needed only when the nonhomologous ends of recombining DNA are 30 nucleotides (nt) long or longer. An additional protein, the helicase Srs2, is required for the RAD1-dependent removal of long 3' tails. We suggest that Srs2 acts to extend and stabilize the initial nascent joint between the invading single strand and its homolog. 3' tails shorter than 30 nt are removed by another mechanism that depends at least in part on the 3'-to-5' proofreading activity of DNA polymerase delta. PMID- 9343446 TI - From the Centers for Disease Control and Prevention. Pneumococcal and influenza vaccination levels among adults aged > or = 65 years--United States, 1995. PMID- 9343447 TI - From the Centers for Disease Control and Prevention. Missed opportunities for pneumococcal and influenza vaccination of Medicare pneumonia inpatients--12 western states, 1995. PMID- 9343448 TI - From the Centers for Disease Control and Prevention. Tuberculosis morbidity- United States, 1996. PMID- 9343449 TI - Public long-term care insurance in Japan. AB - A public long-term care (LTC) insurance program is likely to be introduced to Japan in the year 2000. A consensus on the need for more LTC resources in the rapidly aging society and dissatisfaction with the current system are some of the factors that have contributed to its introduction. Half the costs will be paid by premiums that will be levied on all those older than 40 years, and half will be covered by general taxation. The insurer will be the municipalities with a pooling mechanism at the national level to balance the differences in their demographic structure. The benefits will include institutional care, respite care, day care, home help, visiting nurses, and loan of devices. Eligibility status will be classified into 6 levels that will be determined by assessment of functional and cognitive status. However, there are few mechanisms to limit benefits and contain costs. Problems also exist in the design of the eligibility classification and in the assessment instrument. The proposed LTC insurance system highlights the need for defining what should be included in a "basic package" of LTC as an entitlement for every citizen, for an organizational mechanism and an assessment instrument to deliver services efficiently and equitably, and for physicians to work outside the traditional medical model. To what degree the Japanese public in general, and physicians in particular, is willing to deal with these issues is a challenge for the 21 st century. PMID- 9343450 TI - Outcomes for patients with stroke in managed care vs fee-for-service. PMID- 9343451 TI - Outcomes for patients with stroke in managed care vs fee-for-service. PMID- 9343452 TI - Outcomes for patients with stroke in managed care vs fee-for-service. PMID- 9343453 TI - Detection of group B streptococcal infection. PMID- 9343454 TI - Pharmacologic treatment of alcohol withdrawal. PMID- 9343455 TI - Pharmacologic treatment of alcohol withdrawal. PMID- 9343456 TI - Pharmacologic treatment of alcohol withdrawal. PMID- 9343457 TI - Opiate detoxification under anesthesia. PMID- 9343459 TI - Treatment of delusional parasitoses. PMID- 9343458 TI - Opiate detoxification under anesthesia. PMID- 9343460 TI - Treatment of delusional parasitoses. PMID- 9343461 TI - Death from irreversible pulmonary hypertension associated with short-term use of fenfluramine and phentermine. PMID- 9343462 TI - Occupational therapy for independent-living older adults. A randomized controlled trial. AB - CONTEXT: Preventive health programs may mitigate against the health risks of older adulthood. OBJECTIVE: To evaluate the effectiveness of preventive occupational therapy (OT) services specifically tailored for multiethnic, independent-living older adults. Design.-A randomized controlled trial. SETTING: Two government subsidized apartment complexes for independent-living older adults. SUBJECTS: A total of 361 culturally diverse volunteers aged 60 years or older. INTERVENTION: An OT group, a social activity control group, and a nontreatment control group. The period of treatment was 9 months. MAIN OUTCOME MEASURES: A battery of self-administered questionnaires designed to measure physical and social function, self-rated health, life satisfaction, and depressive symptoms. RESULTS: Benefit attributable to OT treatment was found for the quality of interaction scale on the Functional Status Questionnaire (P=.03), Life Satisfaction Index-Z (P=.03), Medical Outcomes Study Health Perception Survey (P=.05), and for 7 of 8 scales on the RAND 36-Item Health Status Survey, Short Form: bodily pain (P=.03), physical functioning (P=.008), role limitations attributable to health problems (P=.02), vitality (P=.004), social functioning (P=.05), role limitations attributable to emotional problems (P=.05), and general mental health (P=.02). CONCLUSIONS: Significant benefits for the OT preventive treatment group were found across various health, function, and quality-of-life domains. Because the control groups tended to decline over the study interval, our results suggest that preventive health programs based on OT may mitigate against the health risks of older adulthood. PMID- 9343463 TI - A placebo-controlled, double-blind, randomized trial of an extract of Ginkgo biloba for dementia. North American EGb Study Group. AB - CONTEXT: EGb 761 is a particular extract of Ginkgo biloba used in Europe to alleviate symptoms associated with numerous cognitive disorders. Its use in dementias is based on positive results from only a few controlled clinical trials, most of which did not include standard assessments of cognition and behavior. OBJECTIVE: To assess the efficacy and safety of EGb in Alzheimer disease and multi-infarct dementia. DESIGN: A 52-week, randomized double-blind, placebo-controlled, parallel-group, multicenter study. PATIENTS: Mildly to severely demented outpatients with Alzheimer disease or multi-infarct dementia, without other significant medical conditions. INTERVENTION: Patients assigned randomly to treatment with EGb (120 mg/d) or placebo. Safety, compliance, and drug dispensation were monitored every 3 months with complete outcome evaluation at 12, 26, and 52 weeks. PRIMARY OUTCOME MEASURES: Alzheimer's Disease Assessment Scale-Cognitive subscale (ADAS-Cog), Geriatric Evaluation by Relative's Rating Instrument (GERRI), and Clinical Global Impression of Change (CGIC). RESULTS: From 309 patients included in an intent-to-treat analysis, 202 provided evaluable data for the 52-week end point analysis. In the intent-to-treat analysis, the EGbgroup had an ADAS-Cog score 1.4 points better than the placebo group (P=.04) and a GERRI score 0.14 points better than the placebo group (P=.004). The same patterns were observed with the evaluable data set in which 27% of patients treated with EGb achieved at least a 4-point improvement on the ADAS-Cog, compared with 14% taking placebo (P=.005); on the GERRI, 37% were considered improved with EGb, compared with 23% taking placebo (P=.003). No difference was seen in the CGIC. Regarding the safety profile of EGb, no significant differences compared with placebo were observed in the number of patients reporting adverse events or in the incidence and severity of these events. CONCLUSIONS: EGb was safe and appears capable of stabilizing and, in a substantial number of cases, improving the cognitive performance and the social functioning of demented patients for 6 months to 1 year. Although modest, the changes induced by EGb were objectively measured by the ADAS-Cog and were of sufficient magnitude to be recognized by the caregivers in the GERRI. PMID- 9343464 TI - Cost-effectiveness of vaccination against pneumococcal bacteremia among elderly people. AB - CONTEXT: Clinical, epidemiologic, and policy considerations support updating the cost-effectiveness of pneumococcal vaccination for elderly people and targeting the evaluation only to prevention of pneumococcal bacteremia. OBJECTIVE: To assess the implications for medical costs and health effects of vaccination against pneumococcal bacteremia in elderly people. DESIGN: Cost-effectiveness analysis of pneumococcal vaccination compared with no vaccination, from a societal perspective. SETTING AND PARTICIPANTS: The elderly population aged 65 years and older in the United States in 3 geographic areas: metropolitan Atlanta, Ga; Franklin County, Ohio; and Monroe County, New York. MAIN OUTCOME MEASURES: Incremental medical costs and health effects, expressed in quality-adjusted life years per person vaccinated. RESULTS: Vaccination was cost saving, ie, it both reduced medical expenses and improved health, for all age groups and geographic areas analyzed in the base case. For people aged 65 years and older, vaccination saved $8.27 and gained 1.21 quality-adjusted days of life per person vaccinated. Vaccination of the 23 million elderly people unvaccinated in 1993 would have gained about 78000 years of healthy life and saved $194 million. In univariate sensitivity analysis, the results remained cost saving except for doubling vaccination costs, including future medical costs of survivors, and lowering vaccination effectiveness. With assumptions most unfavorable to vaccination, cost per quality-adjusted life-year ranged from $35 822 for ages 65 to 74 years to $598 487 for ages 85 years and older. In probabilistic sensitivity analysis, probability intervals were more narrow, with less than 5% probability that the ratio for ages 85 years and older would exceed $100000. CONCLUSIONS: Pneumococcal vaccination saves costs in the prevention of bacteremia alone and is greatly underused among the elderly population, on both health and economic grounds. These results support recent recommendations of the Advisory Committee on Immunization Practices and public and private efforts under way to improve vaccination rates. PMID- 9343465 TI - Effects of residence in Alzheimer disease special care units on functional outcomes. AB - CONTEXT: Alzheimer disease special care units (SCUs) in nursing homes are increasingly prevalent, but little is known about their effects on residents' outcomes. OBJECTIVE: To analyze the effect of SCU residence on the rates at which residents decline in functional status. DESIGN: A cohort of nursing home residents assessed at multiple points during about 1 year. Facility staff completed all assessments using the Minimum Data Set for Nursing Home Resident Assessment and Care Screening (MDS). SETTING: Medicare- or Medicaid-certified nursing facilities. PATIENTS OR OTHER PARTICIPANTS: All nursing home residents in 1993 and early 1994 in Kansas, Maine, Mississippi, and South Dakota. Serial MDS assessments of 77337 residents in more than 800 facilities, including 1228 residents in 48 facilities with SCUs. MAIN OUTCOME MEASURES: Decline in locomotion, transferring, toileting, eating, dressing, and a summary activities of daily living index; decline in urinary and bowel continence; and significant weight loss. RESULTS: No statistically significant difference was observed in the speed of decline for residents in SCUs and traditional units in any of the 9 outcomes. Residents were matched on a variety of characteristics, and subgroup analyses were performed. In none did we observe a pattern of better outcomes among SCU residents. CONCLUSIONS: Although SCUs may have provided unmeasured benefits to families and residents, it does not appear that those benefits included any slowing in the rates of functional decline experienced by individuals with dementia. PMID- 9343466 TI - Telomeres, cancer, and aging. Altering the human life span. AB - Population projections of the aging global society and its fiscal and social impact have depended on assumptions regarding the human life span. Until now, the assumption that the maximum human life span is fixed has been justified. Recent advances in cell biology, genetics, and our understanding of the cellular processes that underlie aging, however, have shown that this assumption is invalid in a number of animal models and suggest that this assumption may become invalid for humans as well. In vitro alteration of telomeres affects cellular senescence, and in vivo manipulation of genes and diet can increase maximum life span in animal models if these discoveries are extended to humans. We may soon be able to extend the maximum human life span and postpone or prevent the onset of diseases associated with aging. Such a possibility requires that we recognize a growing uncertainty in any attempt to project international health care costs into the next few decades. The costs may be significantly lower than projections, if life span increases and age-related disabilities are postponed or less severe, or perhaps higher, if life span increases without altering the onset and severity of disability. An appropriate uncertainty regarding the human life span undermines any attempt to accurately predict health costs in the next century. PMID- 9343467 TI - Effects of age, sex, and ethnicity on the association between apolipoprotein E genotype and Alzheimer disease. A meta-analysis. APOE and Alzheimer Disease Meta Analysis Consortium. AB - OBJECTIVE: To examine more closely the association between apolipoprotein E (APOE) genotype and Alzheimer disease (AD) by age and sex in populations of various ethnic and racial denominations. DATA SOURCES: Forty research teams contributed data on APOE genotype, sex, age at disease onset, and ethnic background for 5930 patients who met criteria for probable or definite AD and 8607 controls without dementia who were recruited from clinical, community, and brain bank sources. MAIN OUTCOME MEASURES: Odds ratios (ORs) and 95% confidence intervals (CIs) for AD, adjusted for age and study and stratified by major ethnic group (Caucasian, African American, Hispanic, and Japanese) and source, were computed for APOE genotypes epsilon2/epsilon2, epsilon2/epsilon3, epsilon2/epsilon4, epsilon3/epsilon4, and epsilon4/epsilon4 relative to the epsilon3/epsilon3 group. The influence of age and sex on the OR for each genotype was assessed using logistic regression procedures. RESULTS: Among Caucasian subjects from clinic- or autopsy-based studies, the risk of AD was significantly increased for people with genotypes epsilon2/epsilon4 (OR=2.6, 95% CI=1.6-4.0), epsilon3/epsilon4 (OR=3.2, 95% CI=2.8-3.8), and epsilon4/epsilon4 (OR=14.9, 95% CI= 10.8-20.6); whereas, the ORs were decreased for people with genotypes epsilon2/epsilon2 (OR=0.6, 95% CI=0.2-2.0) and epsilon2/epsilon3 (OR=0.6, 95% CI=0.5-0.8). The APOE epsilon4-AD association was weaker among African Americans and Hispanics, but there was significant heterogeneity in ORs among studies of African Americans (P<.03). The APOE epsilon4-AD association in Japanese subjects was stronger than in Caucasian subjects (epsilon3/epsilon4: OR=5.6, 95% CI=3.9 8.0; epsilon4/epsilon4: OR=33.1, 95% CI=13.6-80.5). The epsilon2/epsilon3 genotype appears equally protective across ethnic groups. We also found that among Caucasians, APOE genotype distributions are similar in groups of patients with AD whose diagnoses were determined clinically or by autopsy. In addition, we found that the APOE epsilon4 effect is evident at all ages between 40 and 90 years but diminishes after age 70 years and that the risk of AD associated with a given genotype varies with sex. CONCLUSIONS: The APOE epsilon4 allele represents a major risk factor for AD in all ethnic groups studied, across all ages between 40 and 90 years, and in both men and women. The association between APOE epsilon4 and AD in African Americans requires clarification, and the attenuated effect of APOE epsilon4 in Hispanics should be investigated further. PMID- 9343470 TI - Living longer, contributing longer. PMID- 9343469 TI - Diagnosis and treatment of Alzheimer disease and related disorders. Consensus statement of the American Association for Geriatric Psychiatry, the Alzheimer's Association, and the American Geriatrics Society. AB - OBJECTIVE: A consensus conference on the diagnosis and treatment of Alzheimer disease (AD) and related disorders was organized by the American Association for Geriatric Psychiatry, the Alzheimer's Association, and the American Geriatrics Society on January 4 and 5, 1997. The target audience was primary care physicians, and the following questions were addressed: (1) How prevalent is AD and what are its risk factors? What is its impact on society? (2) What are the different forms of dementia and how can they be recognized? (3) What constitutes safe and effective treatment for AD? What are the indications and contraindications for specific treatments? (4) What management strategies are available to the primary care practitioner? (5) What are the available medical specialty and community resources? (6) What are the important policy issues and how can policymakers improve access to care for dementia patients? (7) What are the most promising questions for future research? PARTICIPANTS: Consensus panel members and expert presenters were drawn from psychiatry, neurology, geriatrics, primary care, psychology, nursing, social work, occupational therapy, epidemiology, and public health and policy. EVIDENCE: The expert presenters summarized data from the world scientific literature on the questions posed to the panel. CONSENSUS PROCESS: The panelists listened to the experts' presentations, reviewed their background papers, and then provided responses to the questions based on these materials. The panel chairs prepared the initial drafts of the consensus statement, and these drafts were read by all panelists and edited until consensus was reached. CONCLUSIONS: Alzheimer disease is the most common disorder causing cognitive decline in old age and exacts a substantial cost on society. Although the diagnosis of AD is often missed or delayed, it is primarily one of inclusion, not exclusion, and usually can be made using standardized clinical criteria. Most cases can be diagnosed and managed in primary care settings, yet some patients with atypical presentations, severe impairment, or complex comorbidity benefit from specialist referral. Alzheimer disease is progressive and irreversible, but pharmacologic therapies for cognitive impairment and nonpharmacologic and pharmacologic treatments for the behavioral problems associated with dementia can enhance quality of life. Psychotherapeutic intervention with family members is often indicated, as nearly half of all caregivers become depressed. Health care delivery to these patients is fragmented and inadequate, and changes in disease management models are adding stresses to the system. New approaches are needed to ensure patients' access to essential resources, and future research should aim to improve diagnostic and therapeutic effectiveness. PMID- 9343468 TI - Taste and smell losses in normal aging and disease. AB - OBJECTIVE: To review the scientific literature on the alterations in the senses of taste and smell in the elderly, including causes, diagnosis, prognosis, and treatment. DATA SOURCES: Original reports and reviews obtained through MEDLINE searches from 1966 through June 1997 using the MeSH headings of "taste," "taste buds," "taste disorders," "taste thresholds," "smell," "odors," "aged," and "aging." Articles frequently cited in reference lists were also included. STUDY SELECTION: All articles were reviewed, tabulated, and summarized. DATA EXTRACTION: Criteria for extraction included data quality and validity, statistical treatment of the data, venue of publication, and relevance to clinical care. CONCLUSION: Losses of taste and smell are common in the elderly and result from normal aging, certain disease states (especially Alzheimer disease), medications, surgical interventions, and environmental exposure. Deficits in these chemical senses cannot only reduce the pleasure and comfort from food, but represent risk factors for nutritional and immune deficiencies as well as adherence to specific dietary regimens. Chemosensory decrements can lead to food poisoning or overexposure to environmentally hazardous chemicals that are otherwise detectable by taste and smell. Use of flavor-enhanced food can increase enjoyment of food and have a positive effect on food intake and immune status. PMID- 9343471 TI - Research on aging. An agenda for all nations individually and collectively. PMID- 9343472 TI - Living longer, aging better. Aging research comes of age. PMID- 9343473 TI - Aging: a global issue. PMID- 9343474 TI - A piece of my mind. Aging and caring. PMID- 9343475 TI - [A computed tomographic study of epilepsy in children: the primary diagnosis of a brain tumor]. AB - There are presented experience of computer tomographic (CT) investigation of 435 children with different epileptic syndromes, which had been verified according to international Classification of Epilepsy and Epileptic Syndromes (New Delhi [correction of New-Daily], 1989). Tumors were diagnosed in 6 cases and were manifested as resistant epileptic fits. There was grounded the necessity of including of the neuroimaging methods in complex study of children with epilepsy for optimization of diagnostics, treatment and prognosing of the course of epilepsy. There were also formulated the practical recommendations concerning carrying out of CT investigations in children with prolonged resistant epileptic syndromes. PMID- 9343476 TI - [Twin studies of myasthenia]. AB - Twin pairs with one or two myasthenic patients were selected from patients with neuromuscular pathology. 18 couples of twins were included in final selection (9 monozygotic and 9 dizygotic pairs). 4 pairs of monozygotic twins were concordant to myasthenia (MA). All dizygotic pairs were discordant to this disease. Matched concordance of monozygotic twins was 44%. Penetration of pathological gene was 61% assuming the hypothesis about monogenic heredity of MA. The coefficient of heredity was 44%. The conclusion was made about important, but not absolute role of hereditary factors in development of MA. It was necessary the presence of combination of both genetic and environmental factors for MA development. PMID- 9343477 TI - [The spectrum of spinal muscular atrophies: a population study]. AB - A population medical genetic study of spinal muscular atrophies (SMA) was carried out in 1800 individuals in 6 russian and 3 middle asian regions. 33 patients with SMA were ascertained including 29 with autosomal recessive childhood proximal SMA (SMA I-III) and 4 patients with rare SMA types. There were revealed either overlapping of diagnostic criteria of I-III types in some patients or interfamilial differences of types in 3 from 6 familial cases, (one of them with distant relatives affected as well as pronounced clinical genetic variability of rare forms of SMA (all sporadic cases). PMID- 9343478 TI - [The treatment of nocturnal enuresis in children]. AB - 367 children with nocturnal enuresis (NE) were divided into randomly selected groups. Two groups were treated with amitriptylin (the 1st group) or imipramine (the 2nd one). In the third group the treatment was differentiated and depended on coexist clinical sleep disturbances. In such cases there were used according to indications amitriptylin (including in combination with cyclodol), imipramine, diazepam. Additionally nootropic drugs (pyracetam, pantogam) were also administrated. In 3 month after the treatment 68.3% of the patients of the 3rd group still had complete remission. These results were better then in the 1st (20.9%) and in the 2nd (45.6%) groups of patients in which the treatment of NE was not depended on coexist disturbances. PMID- 9343479 TI - [Extracorporeal blood correction in the therapy of drug-resistant mental disorders]. AB - 544 patients resistant to pharmacotherapy were treated by different methods of extracorporal hemocorrection (EH). Positive changes in mental state were achieved in more than half of the patients by means of EH without any psychopharmacologic drugs. These changes were caused by antipsychotic and antidepressive effects of EH as well as by its ability to normalize biochemical, immunological and rheological blood indices PMID- 9343480 TI - [Brain bioelectrical activity in patients with the fragile X-chromosome syndrome and in their mothers]. AB - 30 patients (4-22 years old) and their 12 mothers were examined by means of method of electroencephalography (EEG). Healthy individuals of corresponding age were included in the control group. EEG changes of the same type were observed in all the patients with syndrome of fragile X-chromosome: reduction of occipital alpha-rhythm, prevalence of theta-rhythm in central-parietal and central frontal regions as well as epileptoid activity in parietal and central-parietal cortical regions. These peculiarities could be considered as the markers in syndrome's diagnostics. EEG of mothers (heterozygous carriers of mutant gene) were characterised by considerable variations: increase of rolandic rhythm's presentation in central cortical zones, accumulation of hypersynchronous EEG, increase of rhythmical theta-activity, etc. EEG of mother with shift-like schizophrenia resembled one of proband in one case. PMID- 9343481 TI - [A method for determining the mental development of children up to 3 years old- the Schedule for the Neuropsychological Examination of Infants]. AB - GNOM--is standard method of estimation of health state in babies and infants with determination of coefficient of mental development. Method's basis appears to be tasks and tests allowing to evaluate (in scores) the state of the main neuropsychiatric spheres, namely: sensory, motor, emotional, cognitive and social communicative. Method exists both in ordinary and computer variations. It was approved in process of observation of 500 children. GNOM permits to perform the screening of children in big children's contingents. It may be used by either the physicians or other medical personnel as well as by the parents too. PMID- 9343482 TI - [The classification and topical diagnosis of cerebral circulatory disorders in newborn infants]. AB - The classification of disorders of cerebral circulation (DCC) was presented on the basis of clinical-neurosonic study of 100 newborn children with the signs of DCC. There were-determined the same degrees of DCC severity as in hypoxic ischemic encephalopathy (that is light, manifested, severe ones). There weren't observed any signs of focal damages of brain in light form of DCC, although there might sometimes be found the small ischemic damages of brain and injuries of hemorrhagic type in one third of patients. In manifested DCC there were revealed the signs of focal cerebral damages which were caused by combination of hemorrhages in choroid plexus of lateral ventricles and subependymal hemorrhages, and were rarer conditioned by ischemic necrosis of these regions. It was quite characteristic for severe DCC the appearance of pronounced total cerebral disorders just after the birth which might progradiently increase. There were found parenchymatous, subarachnoidal, and intraventricle hemorrhages by means of neurosonography. There was demonstrated the significance of investigation of both muscular tonus and reflexes in newborn children for topic DCC diagnostics. PMID- 9343483 TI - [The Outpatient Pediatric Neurology Service of Moscow today]. AB - The organisation of children's neurological service was considered in terms of 25 years of medical-diagnostic work of children's consultative neurological out patient clinic (CCNC) attached to one of Moscow children's clinical hospital (Morozov hospital). The work of appropriate departments of CCNC was presented together with the consideration of the main problems of child's neurology as well as of the peculiarities of nervous system's pathology in terms of age. The close connection was emphasized between CCNC and child's neurologists of general out patient clinics as well as the role of CCNC both in coordination of their work and in the rise of their qualification. PMID- 9343484 TI - [The mental health status of urban adolescents with nondelinquent and delinquent behaviors]. AB - Mental health was examined in two representative selections of urban adolescents of 15-17 years old: with nondelinquent (n = 624) and delinquent (n = 106) behaviour. The complex of sociomedical, psychological, sociological and statistic methods was applied. High frequency of mental pathology as well as a low level of mental health were revealed in the groups examined, especially in delinquent adolescents. There were determined some differences between the groups in frequencies of separate forms of mental disorders. In nondelinquent adolescents there were most frequently observed borderline mental disorders of subclinical level as well as neuroses; there weren't observed the states of borderline intellectual insufficiency in such individuals. Pathocharacterological personal development, alcoholism, neurotic- and psychopathic-like disorders were more frequently observed in delinquent teenagers. There was determined the leading role of personal psychological behavioral factors in development of considerable mental disorders in adolescents. PMID- 9343485 TI - [The Kearns-Sayre syndrome]. PMID- 9343486 TI - [Lejeune's syndrome]. PMID- 9343487 TI - [Severe myoclonic epilepsy in infancy]. PMID- 9343488 TI - [Horseback riding as a means of treatment and rehabilitation in neurology and psychiatry]. PMID- 9343489 TI - [Depressive states at an early age]. AB - 100 children of 1-3 years old were observed. In 50 of them depressive states developed in conditions of total deprivation--the situation of orphanhood. In other 50 children the depressions were caused by partial deprivation--upbringing by schizophrenic mother (children from group with high risk of schizophrenia). Three variations of depression were found in the first group: autonomic, somatizated, and regressive-apathetic. Depressions in the second group were presented either by infantile depression or by infantile distress-syndrome in frames of schizotypic diathesis. PMID- 9343491 TI - Mechanisms of induction of skin cancer by UV radiation. AB - Ultraviolet (UV) radiation is the carcinogenic factor in sunlight; damage to skin cells from repeated exposure can lead to the development of cancer. UV radiation has been mainly implicated as the cause of non-melanoma skin cancer, although some role for UV in malignant melanoma has been suggested. The induction of skin cancer is mainly caused by the accumulation of mutations caused by UV damage. Cellular mechanisms exist to repair the DNA damage, or to induce apoptosis to remove severely damaged cells; however, the additive effects of mutations in genes involved in these mechanisms, or in control of the cell cycle, can lead to abnormal cell proliferation and tumor development. The molecular events in the induction of skin cancer are being actively investigated, and recent research has added to the understanding of the roles of tumor suppressor and oncogenes in skin cancer. UV radiation has been shown to induce the expression of the p53 tumor suppressor gene, and is known to produce "signature" mutations in p53 in human and mouse skin cancers and in the tumor suppressor gene patched in human basal cell carcinoma. The role of UV radiation in suppression of immune surveillance in the skin, which is an important protection against skin tumor development, is also being investigated. The knowledge gained will help to better understand the ways in which skin cancer arises from UV exposure, which will in turn allow development of better methods of treatment and prevention. PMID- 9343490 TI - Effect of aging and caloric restriction on intestinal sugar and amino acid transport. AB - The incidence of intestinal nutrient malabsorption increases with age. Therefore, an important question is whether there are age-related changes in intestinal nutrient absorption which may contribute to a decline in absorptive capacity. Sugar and amino acid transport per mg intestine generally decreases with age. The proximate mechanism underlying this age-related decrease in transport activity is a decrease in number of transporters per mg. This reduction in transporter number can be caused by age-related changes in cell proliferation rates which, in turn, can alter the ratio of absorptive to nonabsorptive cells. The age-related change in proliferation rates typically increases intestinal mass. There seems to be no age-related changes in the steady state levels of transporter mRNA. Aging also modestly impairs the ability of intestinal nutrient transport systems to adapt to changes in dietary conditions. Caloric restriction is the only procedure known to consistently increase the lifespan of mammals. Chronic caloric restriction markedly enhances intestinal nutrient transport per mg without affecting intestinal mass. Since body weight decreases with caloric restriction, there is a dramatic increase in intestinal absorptive capacity normalized to body weight. This suggests that an increase in intestinal nutrient absorption may be a critical adaptation to caloric restriction. There is a need to perform in vivo transport studies during senescence, to distinguish between acute and chronic effects of caloric restriction, and to identify hormones that may mediate aging and caloric restriction effects on intestinal nutrient transport. PMID- 9343492 TI - The future of asthma. PMID- 9343493 TI - La critique est aisee mais l'art est difficile. [Criticism is easy but the method is hard; comment]. PMID- 9343495 TI - Influence of age on Henoch Schonlein purpura. PMID- 9343494 TI - Non-sedating antihistamines and cardiac arrhythmia. PMID- 9343496 TI - Bisphosphonates for pain relief in reflex sympathetic dystrophy? PMID- 9343497 TI - Diesel fuel and exhaust emissions: is there a human carcinogenic risk? PMID- 9343498 TI - Total cholesterol and risk of mortality in the oldest old. AB - BACKGROUND: The impact of total serum cholesterol as a risk factor for cardiovascular disease decreases with age, which casts doubt on the necessity for cholesterol-lowering therapy in the elderly. We assessed the influence of total cholesterol concentrations on specific and all-cause mortality in people aged 85 years and over. METHODS: In 724 participants (median age 89 years), total cholesterol concentrations were measured and mortality risks calculated over 10 years of follow-up. Three categories of total cholesterol concentrations were defined: < 5.0 mmol/L, 5.0-6.4 mmol/L, and > or = 6.5 mmol/L. In a subgroup of 137 participants, total cholesterol was measured again after 5 years of follow up. Mortality risks for the three categories of total cholesterol concentrations were estimated with a Cox proportional-hazards model, adjusted for age, sex, and cardiovascular risk factors. The primary causes of death were coded according to the International Classification of Diseases (ICD-9). FINDINGS: During 10 years of follow-up from Dec 1, 1986, to Oct 1, 1996, a total of 642 participants died. Each 1 mmol/L increase in total cholesterol corresponded to a 15% decrease in mortality (risk ratio 0.85 [95% CI 0.79-0.91]). This risk estimate was similar in the subgroup of participants who had stable cholesterol concentrations over a 5 year period. The main cause of death was cardiovascular disease with a similar mortality risk in the three total cholesterol categories. Mortality from cancer and infection was significantly lower among the participants in the highest total cholesterol category than in the other categories, which largely explained the lower all-cause mortality in this category. INTERPRETATION: In people older than 85 years, high total cholesterol concentrations are associated with longevity owing to lower mortality from cancer and infection. The effects of cholesterol lowering therapy have yet to be assessed. PMID- 9343499 TI - Effect of organisational downsizing on health of employees. AB - BACKGROUND: Reduction of personnel by businesses and other organisations (organisational downsizing) is common in Europe, but little is known about its effects on the health of employees. METHODS: We used employers' records to investigate the relation between downsizing and subsequent absenteeism because of ill health in 981 local-government workers who remained in employment in Raisio, south-western Finland, during a period of economic decline (1991-95). Data were separated into three time periods: 1991, before downsizing; 1993, major downsizing in some workplaces and occupations; and 1993-95, after downsizing. We obtained data on sick leave from records kept by the occupational health-care unit in Raisio. We also investigated whether the effects of downsizing were dependent on ten other predictors of sick leave. FINDINGS: There was a significant association between downsizing and medically certified sick leave. The rate of absenteeism was 2.3 times greater (95% CI 2.0-2.7) after major downsizing, classified by occupation, than after minor downsizing. The corresponding rate ratios for musculoskeletal disorders and trauma were 5.7 (4.1 8.0) and 2.7 (1.7-4.2), respectively. The effects of downsizing by workplace depended on the age distribution of the staff. When the proportion of employees who were older than 50 years was high, downsizing increased the individual risk of absence because of ill health by 3.2-14.0 times, depending on diagnostic category. When the proportion of employees over 50 years was low, downsizing had only slight effects on health. Other risk factors that increased rates of sick leave after downsizing were age over 44 years, a large workplace, poor health before downsizing, and high income. INTERPRETATION: Downsizing is a risk to the health of employees. But this risk varies according to individual factors, such as age, socioeconomic status, and health, as well as factors related to place of work, for example, size and age structure of the staff. PMID- 9343500 TI - Monitoring the results of cardiac surgery by variable life-adjusted display. AB - BACKGROUND: Conventional assessment of the outcome of cardiac surgery usually takes the form of retrospective mortality figures and, at best, indicates an average performance over time. Summary tables conceal good and bad runs, and without risk adjustment they are difficult to interpret. We developed a refinement of the cumulative sum method that weights death and survival by each patient's risk status and provides a display of surgical performance over time. METHODS: The variable life-adjusted (VLAD) plot shows the difference between expected and actual cumulative mortality. VLAD shows whether a surgeon's performance is above or below what might be expected. This mortality-scoring system accumulates penalties for each death and rewards for every survivor, based on the inherent risk of perioperative death of each case concerned. FINDINGS: We illustrate the results of three performance reviews, displayed as VLADs. The first shows the results of an individual surgeon for 547 consecutive cardiac surgical cases. The overall mortality was 36% less than that predicted by the Parsonnet scoring system. The second displays the results for 5000 consecutive patients who underwent cardiopulmonary bypass between 1992 and 1996, divided into six contemporaneous series. The predicted mortality was 9% compared with 6% actual mortality. The third is a plot for a trainee surgeon and clearly shows how a period of poor performance was identified and then substantially improved, which would not have been revealed by conventional tables of summary statistics. INTERPRETATION: VLAD provides a graphical display of risk-adjusted survival figures for individual surgeons or units over time and could be modified to monitor performance over a range of treatments and outcomes. PMID- 9343501 TI - Fetal growth, length of gestation, and polycystic ovaries in adult life. AB - BACKGROUND: Polycystic ovaries are a common disorder associated with menstrual irregularities, subfertility, hirsutism, acne, and a range of endocrine abnormalities, including high concentrations of plasma luteinising hormone (LH) and excessive androgen production. The pathophysiology is not understood. We investigated whether the disorder originates during intrauterine life. METHODS: We examined 235 women aged 40-42 years who were born in Sheffield, UK. We related the prevalence of polycystic ovaries and the plasma concentrations of gonadotropin hormones and androgens to the women's body size at birth, and the length of gestation. FINDINGS: 49 (21%) of the women had polycystic ovaries. We defined two groups of women with the disorder, which correspond to the two groups that commonly present clinically. The first group comprised obese women who were androgenised, with higher than normal concentrations of plasma LH and testosterone. These women had above-average birthweight and were born to overweight mothers. The second group comprised women of normal weight who had high plasma LH, but normal testosterone concentrations. These women were born after term (40 weeks' gestation). INTERPRETATION: The two common forms of polycystic ovary syndrome have different origins in intrauterine life. Obese, hirsute women with polycystic ovaries have higher than normal ovarian secretion of androgens that are associated with high birthweight and maternal obesity. Thin women with polycystic ovaries have altered hypothalamic control of LH release resulting from prolonged gestation. PMID- 9343502 TI - Association of slow acetylator genotype for N-acetyltransferase 2 with familial Parkinson's disease. AB - BACKGROUND: Epidemiological studies have identified positive family history and exposure to environmental toxins as risk factors for Parkinson's disease (PD). An inherited defect of xenobiotic metabolism could result in increased susceptibility to such toxins. We investigated the frequency of functionally relevant polymorphisms in six detoxification enzymes among patients with PD to elucidate the relation between these polymorphisms and the disease. METHODS: We obtained brain-tissue samples from 100 patients with apparently sporadic PD and blood samples from 100 living patients with familial PD. For the control group, we extracted DNA from the tissue of 100 pathologically normal brains. The six enzymes analysed in these three groups were: CYP2D6, CYP2E1, NAD(P)H-menadione reductase, glutathione transferases M1 and T1, and N-acetyltransferase 2. We also investigated N-acetyltransferase 2 in 100 blood samples from patients with genetically proven Huntington's disease. We used PCR-based methods and restriction-enzyme analysis to detect polymorphisms. FINDINGS: The slow acetylator genotype for N-acetyltransferase 2 was more common in the familial PD group (69%) than in all controls (37%). Even after correction for multiple comparisons, this result remained highly significant (p = 0.002) for familial PD compared with normal controls (odds ratio 3.79 [95% CI 2.08-6.90]) and compared with Huntington's disease (2.45 [1.37-4.38], p = 0.004). The slow acetylator frequency for N-acetyltransferase 2 for sporadic PD was between that for Huntington's disease and familial PD. The frequencies of all the other polymorphisms were similar in the two study groups and the normal control group. INTERPRETATION: We found an association between the slow acetylator genotype for N-acetyltransferase 2 and familial PD. Further studies are needed to investigate the biological relevance of these findings, but slow acetylation could lead to impaired ability of patients with familial PD to handle neurotoxic substances. PMID- 9343503 TI - Lightheaded spells and hypertension. PMID- 9343504 TI - HIV-1 in semen: an isolated virus reservoir. PMID- 9343505 TI - Bioengineered skin. PMID- 9343506 TI - Increase in primary liver cancer in the UK, 1979-94. PMID- 9343507 TI - Safety of first-trimester exposure to topical tretinoin: prospective cohort study. PMID- 9343509 TI - HHV-8 and multiple myeloma in the UK. PMID- 9343508 TI - HHV-8 and multiple myeloma in France. PMID- 9343510 TI - Migraine and cerebral blood flow during centrifugation. PMID- 9343511 TI - Combined endovascular and surgical treatment of complex traumatic lesions of thoracic aorta. PMID- 9343512 TI - Moi scuppers sex-education plans in Kenya. PMID- 9343514 TI - The omnipresent still syndrome. PMID- 9343513 TI - Canadian Red Cross found negligent. PMID- 9343515 TI - How should clinical care of the aged differ? PMID- 9343516 TI - Dying, not old age, to blame for costs of health care. PMID- 9343517 TI - Myths of ageing. PMID- 9343518 TI - Access to advances in cardiology. PMID- 9343519 TI - Living a little more dangerously. PMID- 9343520 TI - Is the best yet to be? PMID- 9343521 TI - Added years, onus or bonus? PMID- 9343522 TI - Fall-induced injuries among elderly people. PMID- 9343523 TI - Statins and hypercholesterolaemia: UK Standing Medical Advisory Committee guidelines. PMID- 9343524 TI - Artificial neural networks. PMID- 9343525 TI - Effect of a disulfiram metabolite on retinaldehyde metabolism. PMID- 9343526 TI - Penicillin-resistant pneumococcal meningitis. PMID- 9343527 TI - Regional ventilation in the prone position. PMID- 9343528 TI - CARS trial: warfarin and thrombin generation. Coumadin Aspirin Reinfarction Study. PMID- 9343529 TI - Serum theophylline and menstrual asthma attack. PMID- 9343530 TI - Multiple sclerosis as a diagnosis. PMID- 9343531 TI - Radical surgery versus radiotherapy for stage Ib-IIa cervical cancer. PMID- 9343532 TI - Radical surgery versus radiotherapy for stage Ib-IIa cervical cancer. PMID- 9343533 TI - HIV-1 and AIDS awareness in a marginalised Namibian community. PMID- 9343534 TI - Malaria catastrophe in east Africa. PMID- 9343535 TI - Keeping patients informed. PMID- 9343536 TI - Meta-analyses of randomised controlled trials. PMID- 9343537 TI - Meta-analyses of randomised controlled trials. PMID- 9343538 TI - Nutrition, biosecurity key components to healthy bulls. PMID- 9343540 TI - Creativity and innovation for food animal veterinarians. PMID- 9343539 TI - Recent findings in study of papillomatous digital dermatitis. PMID- 9343541 TI - What is your diagnosis? Osteosarcoma of the spine in a dog. PMID- 9343542 TI - Geographic distribution of female and male veterinarians in the United States, 1996. PMID- 9343543 TI - Use of urine cortisol:creatinine ratio versus adrenocorticotropic hormone stimulation testing for monitoring mitotane treatment of pituitary-dependent hyperadrenocorticism in dogs. AB - OBJECTIVE: To evaluate use of urine cortisol-to-creatinine ratio (UC:C) as a means of monitoring response to long-term mitotane treatment in dogs with pituitary-dependent hyperadrenocorticism. DESIGN: Prospective uncontrolled study. ANIMALS: 101 dogs with pituitary-dependent hyperadrenocorticism. PROCEDURE: Urine samples were obtained from dogs on the morning an ACTH stimulation test was performed, and owners were asked their opinion on the health of their dog to monitor response to mitotane treatment. Urine was assayed for cortisol and creatinine concentrations, and UC:C was calculated. The UC:C was compared with post-ACTH plasma cortisol concentration. RESULTS: Post-ACTH plasma cortisol concentration was used to categorize each dog's response to mitotane treatment. The UC:C did not correlate satisfactorily with results of ACTH stimulation testing. Twenty-seven of 85 (32%) dogs would have been incorrectly considered as having received appropriate doses using UC:C. In addition, 16 dogs that received overdoses could not be distinguished from 29 dogs that received appropriate doses. CLINICAL IMPLICATIONS: UC:C does not provide a consistent, correct assessment of mitotane-induced adrenocortical destruction. The ACTH stimulation test, although more time-consuming and expensive, is recommended for monitoring response to mitotane treatment. PMID- 9343544 TI - Necrotizing encephalitis in a Yorkshire terrier. PMID- 9343545 TI - Open reduction and bone plate stabilization, compared with closed reduction and external fixation, for treatment of comminuted tibial fractures: 47 cases (1980 1995) in dogs. AB - OBJECTIVE: To compare open reduction and bone plate fixation with closed reduction and external skeletal fixation as treatment for severely comminuted fractures of the tibia. Limb alignment, fracture reduction, operating time, hospitalization time, postoperative care, time to unrestricted activity, bone healing, complications, and number of surgical procedures were considered. DESIGN: Retrospective case series. ANIMALS: 47 dogs with severely comminuted fractures of the tibia treated with open reduction and bone plate application (22 dogs) or closed reduction and external fixation (25 dogs). PROCEDURE: Medical records of all dogs included in this study were reviewed. Postoperative and follow-up radiographs were evaluated by 2 independent observers. RESULTS: Differences were not found in hospitalization time, time to unrestricted activity, or time to earliest radiographic evidence of bone healing between dogs with fractures treated with a bone plate and dogs with fractures treated with an external fixator. Fractures treated with an external fixator had more caudal malalignment, and fractures treated with a bone plate had more valgus malalignment. Malalignments were determined not to be related to clinical problems. Dogs with fractures treated with an external fixator had shorter surgery times and more recheck examinations. Dogs with fractures treated with a bone plate had more complications. CLINICAL IMPLICATIONS: Open reduction with bone plate fixation and closed reduction with external fixation were both effective for treatment of comminuted tibial fractures. External fixation was associated with shorter surgery time, but dogs required more extensive postoperative care. Bone plate fixation was associated with more complications. PMID- 9343546 TI - Small intestinal fibrosis in two horses. PMID- 9343547 TI - Clinical features of blister beetle poisoning in equids: 70 cases (1983-1996). AB - OBJECTIVE: To document clinical signs and gross pathologic changes associated with naturally acquired cantharidiasis (blister beetle poisoning) in equids. DESIGN: Retrospective study. ANIMALS: 70 equids with laboratory-confirmed blister beetle poisoning. PROCEDURE: Medical records were reviewed to obtain history, physical examination findings, feeding practices, and diagnostic test and necropsy results. RESULTS: 32 horses and 2 donkeys died from exposure to cantharidin, whereas 36 horses survived. Diet content varied, but alfalfa hay was the common component. Onset of signs of disease was rapid. Most equids had signs of gastrointestinal tract distress. Six horses had nonspecific neurologic signs. All equids dying from cantharidiasis were in shock terminally, with duration of clinical signs ranging from 3 to 18 hours. Six horses that died had no gross lesions, whereas 14 had mild to moderate erythema of gastric, small intestinal, or colonic mucosa. Only 2 horses had gastric or duodenal ulceration, and 2 had hemorrhage of the urinary bladder mucosa. One horse had cardiac muscle necrosis. Clinicopathologic data available on 10 horses included hypocalcemia, hypomagnesemia, and azotemia. Cantharidin concentrations in urine or pooled gastric-cecal contents did not always correlate with severity of disease. CLINICAL IMPLICATIONS: Blister beetle poisoning is not universally fatal in equids. Clinical signs are related to the amount of cantharidin ingested. Every horse that survived was treated aggressively. In fatal poisonings, gross lesions may be minimal or inapparent, and diagnosis must be confirmed by chemical detection of cantharidin in urine, blood, or stomach or cecal contents. PMID- 9343548 TI - Epiglottic augmentation for treatment of dorsal displacement of the soft palate in racehorses: 59 cases (1985-1994). AB - OBJECTIVE: To determine whether epiglottic augmentation, in conjunction with more traditional surgical methods, would be useful in the treatment of dorsal displacement of the soft palate in racehorses. DESIGN: Retrospective study. ANIMALS: 40 Thoroughbred and 19 Standardbred racehorses. PROCEDURE: Polytetrafluoroethylene paste was injected submucosally on the lingual epiglottic surface of each horse. In addition, sternothyrohyoideus myectomy or sternothyroideus tenectomy and staphylectomy were performed in most horses. RESULTS: Racing performance was improved after surgery in 29 of 40 (73%) Thoroughbreds and 10 of 19 (53%) Standardbreds. Twenty-nine (49%) horses won > or = 1 race after surgery. CLINICAL IMPLICATIONS: Results suggest that epiglottic augmentation, in conjunction with other surgical methods, may be an effective method of treating horses with poor racing performance attributable to dorsal displacement of the soft palate. PMID- 9343549 TI - Number of viable bacteria and presumptive antibiotic residues in milk fed to calves on commercial dairies. AB - OBJECTIVE: To assess the number of bacteria and presumptive antibiotic residues in milk fed to calves and to identify those bacteria and the antibiotic susceptibility of selected bacterial strains. DESIGN: Cross-sectional prospective study. SAMPLE POPULATION: 189 samples obtained from 12 local dairies. PROCEDURE: Samples of waste milk and milk-based fluids (eg, milk replacer, colostrum, bulk tank milk) were obtained. Cumulative number of viable bacteria was determined. Bacteria were cultured aerobically, and antibiotic susceptibility testing of selected strains was performed. Presumptive antibiotic residues were detected by use of test kits. RESULTS: Geometric mean of the cumulative number of bacteria for waste milk samples was significantly higher than for other types of milk or milk-based products. Streptococcus sp (84/165 samples) and Enterobacteriaceae (83/165 samples) were the predominant bacteria identified, followed by Staphylococcus sp (68/165 samples). Escherichia coli was the gram-negative species most commonly isolated (52/165 samples; 32%); however, none were strain O157. Salmonella sp or Mycoplasma sp were not isolated. Of 189 samples, 119 (63%) were positive when tested for beta-lactams or tetracycline by use of 2 commercially available assays. In vitro, some bacteria were resistant to commonly used antibiotics. CLINICAL IMPLICATIONS: Waste milk that has not been effectively treated (eg, pasteurization) to reduce microbial load prior to use as calf feed should be used with caution, because it may contain a high number of bacteria that may be pathogenic to cattle and human beings. Antibiotic residues that would constitute violative amounts and existence of multiple antibiotic resistant bacterial strains are concerns in calf health management and dairy food safety. PMID- 9343550 TI - Epidemiologic analysis of Mycoplasma spp isolated from bulk-tank milk samples obtained from dairy herds that were members of a milk cooperative. AB - OBJECTIVE: To determine the prevalence of Mycoplasma spp in herds that were members of a milk cooperative. DESIGN: Epidemiologic study. SAMPLE POPULATION: 267 dairy herds that were members of a milk cooperative. PROCEDURE: Bulk-tank milk samples were collected monthly during a 6-year period from all dairies in the cooperative. Samples were submitted to the cooperative's laboratory for bacterial culture for Mycoplasma spp, using direct plating. Milk samples positive for Mycoplasma organisms were speciated. RESULTS: Prevalence of positive samples varied from 1.8 to 5.8% for all species of Mycoplasma and from 1.2 to 3.1% for Mycoplasma spp known to be mastitis pathogens. One mycoplasmal species was isolated initially on 99 of 198 (50.0%) dairies, but 68 of 198 (34.3%) dairies had 2 species isolated. Mycoplasma bovis, M californicum, and M bovigenitalium were consistently isolated, but M bovis (243/499; 48.6%) was the most commonly isolated species. Acholeplasma laidlawii was more prevalent in 1989 and 1995 than other years. Mycoplasma bovigenitalium and M californicum had a seasonal distribution. Less than 50 colonies per plate were isolated for most (317/500; 63.4%) bulk-tank samples. Of the milk samples with > 100 colonies/plate, Mycoplasma bovis was isolated most frequently (73/243; 30.0%). CLINICAL IMPLICATIONS: Distribution of Mycoplasma spp varied by year, number of colonies isolated per sample, season, and herd. Therefore, it may be necessary to routinely sample bulk-tank milk, and all isolates should be speciated. Culture results from milk cooperatives should be used with other monitoring information to determine the Mycoplasma status of herds. PMID- 9343551 TI - Effects of water supplementation with selenium and vitamin E on growth performance and blood selenium and serum vitamin E concentrations in weanling pigs. AB - OBJECTIVE: To determine effects of supplementation of drinking water with selenium and vitamin E on blood selenium and serum vitamin E concentrations, growth performance, and water intake of pigs. DESIGN: Prospective controlled study. ANIMALS: 228 weanling pigs. PROCEDURE: In experiments 1 and 2, pigs were given drinking water supplemented with selenium and vitamin E, and blood selenium and serum vitamin E concentrations were measured. In experiment 3, growth performance and water intake were measured in pigs that received supplemented water for 2 or 5 weeks and in control pigs. RESULTS: In experiment 1, blood selenium concentrations were significantly increased after 7 days of supplementation, and serum vitamin E concentrations were significantly increased after 1 and 7 days of supplementation, compared with baseline concentrations. In experiment 2, blood selenium concentrations were not significantly different between treated and control pigs, and serum vitamin E concentrations were significantly increased on day 7. In experiment 3, gain-to-feed ratios were significantly higher for pigs supplemented with selenium and vitamin E for 5 weeks, but other differences were not detected. CLINICAL IMPLICATIONS: Supplementation of drinking water with selenium and vitamin E may improve the selenium and vitamin E status of weanling pigs by increasing selenium and vitamin E intake. PMID- 9343552 TI - Neurologic examination of sea turtles. AB - OBJECTIVE: To determine whether neurologic examination techniques established for use on dogs and cats could be adapted for use on sea turtles. DESIGN: Prospective controlled observational study. ANIMALS: 4 healthy Green Turtles (Chelonia mydas), 1 healthy Kemp's ridley sea turtle (Lepidochelys kempi), and 6 Green Turtles suspected to have neurologic abnormalities. PROCEDURE: Neurologic examinations were performed while sea turtles were in and out of the water and in ventral and dorsal recumbency. Mentation, general activity, head and body posture, movement and coordination, thoracic and pelvic limb movement, strength and muscle tone, and tail movement were observed. Thoracic and pelvic limb flexor reflexes and nociception, righting response, cranial nerve reflexes, clasp and cloacal reflexes, and neck, dorsal scute, cloacal and tail nociception were tested. RESULTS: Results of neurologic evaluations were consistent for healthy sea turtles. Sea turtles suspected to have neurologic abnormalities had abnormal results. CLINICAL IMPLICATIONS: Many of the neurologic examination techniques used to evaluate dogs and cats can be adapted and used to evaluate sea turtles. A standardized neurologic examination should result in an accurate assessment of neurologic function in impaired sea turtles and should help in evaluating effects of rehabilitation efforts and suitability for return to their natural environment. PMID- 9343553 TI - Developmental changes in tritium autoabsorption. AB - Developmental changes in autoabsorption of tritium emissions were examined in 30 brain regions in the rat at Postnatal Days 0, 4, 7, 10, 14, 21, and 28 and adulthood. Rats received tritiated 2-deoxyglucose in vivo. Alternate brain sections were extracted in chloroform, and autoradiographs were developed from extracted and nonextracted sections. The ratio of optical density values in extracted vs nonextracted sections was used to determine autoabsorption for each structure. Three principal temporal patterns in the development of adult levels of autoabsorption, determined by the optical density ratios, were identified: (1) a minimal increase pattern in which autoabsorption rose only slightly between birth and adulthood; (2) a plateau pattern in which a rapid early increase was followed by stable values; and (3) a late increase pattern in which autoabsorption remained relatively constant until Postnatal Day 28, with a large increase between Day 28 and adulthood. In addition, optical density ratios fluctuated during the second postnatal week in close to one-third of the structures. The data suggest that developmental events affecting the ratio of gray to white matter produce substantial local variations in the development of adult levels of autoabsorption that are distinct for each structure. To correct for autoabsorption effects in ontogenetic studies using tritium autoradiography, it is necessary to determine directly the degree of autoabsorption at a particular time point for the structure of interest. Our results indicate that the technique of in vivo administration of tritiated 2-deoxyglucose followed by chloroform extraction appears to be a sensitive and reproducible method for assessing autoabsorption at all ages. PMID- 9343555 TI - Spatial overlap of rubrospinal and corticospinal terminals with input to the inferior olive. AB - Somatosensory responses of cells in the dorsal accessory olive are suppressed following stimulation of the magnocellular red nucleus. Since the magnocellular red nucleus of the cat does not project directly to the dorsal accessory olive, the present experiments were designed to identify indirect pathways that might mediate suppression of olivary responsiveness. Wheat germ agglutinin-horseradish peroxidase was used to compare the location of magnocellular red nucleus terminals with the locations of cells providing input to the rostral dorsal accessory olive. Cells projecting to forelimb rostral dorsal accessory olive can be divided into two main groups: one group comprises a column of large cells located in the ventral caudal cuneate nucleus extending into lamina VI of C1 and C2, and a second group comprises smaller cells located in the ventral rostral cuneate nucleus. Terminations of fibers originating in the magnocellular red nucleus were found to target both groups of cells projecting to the dorsal accessory olive. Therefore, it is possible that the responsiveness of olivary cells is influenced via these terminations. Stimulation of sensorimotor cortex has also been shown to inhibit olivary responsiveness. Terminations from sensorimotor cortex target the same regions of cells that project to the dorsal accessory olive as those of the magnocellular red nucleus, and a similar, perhaps identical, anatomical substrate may serve to modulate olivary sensitivity by the two descending systems. PMID- 9343554 TI - Measurement of neuronal surface area using high-voltage electron microscope tomography. AB - High-voltage electron microscopy (HVEM) and axial tomography were used to reconstruct the three-dimensional structure of dendrites of intracellularly stained neostriatal spiny neurons. Neurons were stained using iontophoretic injection of horseradish peroxidase from an intracellular micropipet. Thick sections were cut on a vibratome, reacted with diaminobenzidine, and embedded in plastic. High-voltage electron microscopy was used to obtain high-resolution images of neuronal processes in 2- to 3-micron plastic sections cut from those blocks. Series of high-voltage electron micrographs were taken at 2 degrees increments of specimen tilt over a range of at least +/- 60 degrees, and three dimensional reconstructions were generated from the series using an R-weighted backprojection method. An interactive procedure was used to draw the dendritic profiles from slices through the reconstructed volume and to measure volumes and surface areas of the dendrites. Values obtained in this manner matched previous findings using reconstruction from serial thin sections. The HVEM-tomography method offers an alternative to the serial-thin-section method for the quantitative analysis of neuronal shape. PMID- 9343556 TI - Anatomical mapping of functional activation in stereotactic coordinate space. AB - Numerous applications have been reported for the stereotactic mapping of focal changes in cerebral blood flow during sensory and cognitive activation as measured with positron emission tomography (PET) subtraction images. Since these images lack significant anatomical information, analysis of these kinds of data has been restricted to an automated search for peaks in the PET subtraction dataset and localization of the peak coordinates within a standardized stereotactic atlas. This method is designed to identify isolated foci with dimensions smaller than the image resolution. Details of activation patterns that may extend over finite distances, following the underlying anatomical structures, will not be apparent. We describe the combined mapping into stereotactic coordinate space of magnetic resonance imaging (MRI) and PET information from each of a set of subjects such that the major features of the activation pattern, particularly extended tracts of increased blood flow, can be immediately assessed within their true anatomical context as opposed to that presumed using a standard atlas alone. Near areas of high anatomical variability, e.g., central sulcus, or of sharp curvature, e.g., frontal and temporal poles, this information can be essential to the localization of a focus to the correct gyrus or for the rejection of extracerebral peaks. It also allows for the removal from further analysis of data from cognitively-normal subjects with abnormal anatomy such as enlarged ventricles. In patients with neuropathology, e.g., Alzheimer's disease, arteriovenous malformation, stroke, or neoplasm, the use of correlated MRI is mandatory for correct localization of functional activation. PMID- 9343557 TI - Programs for visualization in three-dimensional microscopy. AB - Three-dimensional data representing biological structures can be derived using several methods, including serial section reconstruction, optical sectioning, and tomography. The investigation, comprehension, and communication of structural relationships to others is greatly facilitated by computer-based visualization procedures. We describe SYNU, a suite of programs developed for interactive investigation of three-dimensional structure and for the production of high quality three-dimensional images and animations. We illustrate the capabilities of SYNU in applications to biological data obtained by confocal light microscopy, serial section, and high-resolution electron microscopy from investigations at the cellular, subcellular, and molecular levels. PMID- 9343558 TI - A comparative fractal analysis of various mammalian astroglial cell types. AB - Camera-lucida drawings of Golgi-impregnated astroglial cells and their processes are described by the fractal dimension of their borders, which is an objective, quantitative measure of morphological complexity. Protoplasmic astrocytes from human neocortex have fractal dimensions (D) that are larger than those of fibrous astrocytes from the cat optic nerve. Marginal astrocytes from monkey cerebropontile angle have two kinds of processes: (1) short, thick processes with endfeet abutting the pial surface, with relatively high D's, and (2) very long, thin processes extending into the neuronal tissue, with very low D's. These data indicate that short astrocytic processes may have a complex surface (and have a high D), whereas long processes are rather smooth (and have a low D). A comparison between transmission electron microscopy morphometry and measures of D at the light microscopic level, performed on different parts of rabbit retinal Muller glial cells, suggests that D is strongly correlated to the surface-to volume ratio which, in part, determines the length constant of a cable for core conductance of currents. We provide data supporting the hypothesis that astroglial cell geometry is adjusted to allow for sufficient spatial buffering K+ currents, even through very long processes. PMID- 9343559 TI - Computerized three-dimensional reconstruction reveals cerebrovascular regulatory subregions in rat brain stem. AB - Three-dimensional wireframe reconstructions were used to examine the relationship between the anatomical localization of electrode sites and the cerebrovascular response which was elicited by electrical stimulation of the dorsal raphe nucleus (DRN). Reconstructions of the rat brain and DRN were done from atlas plates and from Nissl-stained coronal sections (100-micron increments). Data points were entered and three-dimensional reconstructions were performed using commercially available software and a personal computer. Display of the entire brain yielded views which obscured visualization of the DRN. The data file was edited to reduce the number of contours without affecting the display resolution of the DRN. Selective display of the DRN and electronic rotation from the coronal to a sagittal view revealed a functional organization of the cerebral blood flow responses which was not apparent in two-dimensional coronal sections. PMID- 9343560 TI - Real complete three-dimensional reconstruction of Golgi-impregnated neurons by means of a confocal laser scanning microscope. AB - Golgi-impregnated neurons of the human or animal central nervous system were studied with a confocal laser scanning microscope (CLSM). The scanning properties (optical sectioning of the specimen) offered by the CLSM and the capacity of metal granules to reflect the laser beam allow a three-dimensional reconstruction of the impregnated neurons. The volume scanned can be depicted in three different ways: (a) extended focus, i.e., a bidimensional image that contains information from all the optical sections, as if there were an extensive depth of focus; (b) a topographic representation in which the intensity of every pixel is proportional to the calculated z value (as a result, the closer the object section is to the surface, the greater the color intensity becomes); and (c) shadow representation, i.e., a pseudo-three-dimensional image. In addition, a true and complete three-dimensional reconstruction of neurons can be obtained using an extended RAM and quick elaboration (fast CPU) combined with the rotation of the reconstructed image in the different planes. High-magnification, high numerical-aperture (NA) oil immersion objective lenses with reduced working distance may present some problems in the three-dimensional reconstruction of large neurons with extensive and spreading dendritic branches. This limitation may be overcome by using a low-magnification (10 x) oil immersion lens. PMID- 9343561 TI - Measurements of motoneuron somal volumes using laser confocal microscopy: comparisons with shape-based stereological estimations. AB - Previous studies on motoneuronal morphometry have assumed the geometry and orientation of neuronal soma in estimating somal volumes based on two-dimensional measurements. In this study, the optical sectioning property of the confocal microscope was used to make direct measurements of phrenic moto-neuron somal volume. These measured volumes were compared to shape-based stereological estimates of volume. Phrenic motoneuron pools in adult rats were retrogradely labeled with fluorescent dye. Labeled motoneurons, in 150-micron-thick tissue sections, were imaged using a Bio-Rad MRC 500 confocal microscope. Somal volumes were directly measured using ANALYZE, a comprehensive image processing software package. These volumes were compared to volume estimates based on five geometrical shapes previously used to study spinal motoneurons. In the adult phrenic motoneuron pool, overestimations of somal volume up to 300% were observed in cases where the Z-axis dimensions of the neurons were not simply related to the X and Y dimensions. None of the five geometrical shapes were found to be suitable for estimating either mean or individual somal volumes of the phrenic motoneuron pool. Confocal microscopy allowed accurate reconstruction along X, Y, and Z axes, and therefore provided a more direct method of measuring motoneuron somal volumes. We conclude that significant and inconsistent errors can be introduced by using shape-based stereological methods for estimating neuronal somal volumes. PMID- 9343562 TI - Optical imaging of cytosolic calcium, electrophysiology, and ultrastructure in pyramidal neurons of organotypic slice cultures from rat hippocampus. AB - Organotypic slice cultures from rat hippocampal cortex grown in an interface between culture medium and a CO2-enriched atmosphere maintained much of the morphological connectivity characteristic of the hippocampus in situ and thinned out considerably, facilitating optical measurements of fluorescent dyes sensitive to Ca2+ in individual neurons. Pyramidal neurons of the CA3 region presented morphological features of differentiated cells, including complex dendritic arborization and large numbers of dendritic spines. The fine cytoskeletal substructure at the postsynaptic density, below the plasma membrane, and within the core of the head and neck of dendritic spines in rapidly frozen slice cultures presents the characteristic morphology previously described for Purkinje cell dendritic spines in acutely dissected cerebellar cortex slices after rapid freezing. CA3 neurons responded to intracellular current injection with a train of action potentials, spike frequency adaptation, and a slow afterhyperpolarization. These spike trains caused rapid increases in dendritic [Ca2+]i that decayed to resting levels after termination of the current pulse. Dendritic spines were clearly observed in proximal dendrites of CA3 neurons in live preparations. [Ca2+]i transients in these dendritic spines closely followed the changes observed in the main dendritic shaft. Orthodromic synaptic stimulation from the dentate hilus generated long-lasting synaptic potentials accompanied by large [Ca2+]i transient in CA3 pyramidal neurons. The [Ca2+]i response was first observed in the proximal dendrites, after which the soma exhibited a [Ca2+]i increase, returning to resting levels within 10 s after the synaptic stimulus. Slice cultures thus provide a favorable opportunity to investigate [Ca2+]i responses in individual neurons maintained in an organotypic synaptic environment, taking advantage of high-resolution optical techniques. PMID- 9343563 TI - Novel method for stereo imaging in light microscopy at high magnifications. AB - A new method for realizing direct stereoscopic (3D) views of thick microscopic sections employs multiple oblique illuminating beams and a single objective lens. Excellent 3D images are obtained in the higher magnification range, where conventional stereo microscopes no longer function. Using conventional microscope optics, significant increases in depth of focus and sharpness are demonstrated. PMID- 9343565 TI - Imaging F-actin in fixed glial cells with a combined optical fluorescence/atomic force microscope. AB - A prototype combined optical fluorescence/atomic force microscope (OFAFM) designed for use in neurobiology and related disciplines has been constructed and used to study filamentous actin (F-actin) and other cellular structures in fixed Xenopus retinal glial cells (XR1 glial cell line). F-actin was readily observed by both fluorescence and AFM. AFM images of nuclei and other cellular structures were also obtained. The OFAFM consists of an AFM with an interferometer detection mechanism mounted on an inverted optical microscope. Integration of optical and scanned probe imaging methods provides a unique and useful approach to studying glial (and other) cell structure and function. PMID- 9343564 TI - Pathfinding by neuroblastoma cells in culture is directed by preferential adhesion to positively charged surfaces. AB - Pathfinding is a fundamental behavior of migrating neuroblasts and advancing growth cones. We have analyzed this behavior in culture using mouse neuroblastoma (N1E-115) cells grown on a chemically patterned surface. The patterned surface was defined photolithographically and consisted of intersection 10-micron-wide pathways. The pathways were coated with positively charged amines and separated by regions bound with uncharged alkanes. Cells and growth cones were guided along the pathways and made choices at intersections. Whereas migrating cells made random choices at intersections, growth cones displayed a preference for advancing straight ahead. Interference reflection microscopy (IRM) revealed that pathfinding by cells and growth cones was correlated with greater overall attachment to aminated regions, although cell bodies and appendages also attached to adjacent alkanated regions. Thus guidance was not simply due to contact inhibition by alkanes; rather, it was due to "preferential" adhesion to aminated surfaces. Gray level analysis of IRM images demonstrated that focal and close contacts were made on both surfaces, indicating that preferential adhesion was not the result of tighter attachment to aminated surfaces. Fluorescent labeling of F-actin and microtubules indicated that preferential adhesion was not due to compartmentalization of these cytoskeletal structures on aminated regions. We propose that preferential adhesion involved a signal transduction mechanism that discriminated between positively charged and uncharged molecules. Such a mechanism could contribute to pathfinding by neuroblasts and growth cones along extracellular matrix proteins in vivo. PMID- 9343566 TI - Detection and estimation of mRNA levels using a nonlinear model in neurons labeled by in situ hybridization histochemistry. AB - In situ hybridization histochemistry (ISHH) is an anatomical technique used to monitor gene expression at the cellular level via detection of steady-state levels of mRNA. Previously, densitometric analysis of ISHH-generated autoradiographic material has provided a relatively quantitative measure of the level of a specific mRNA distributed in any given anatomical region. The present study details the development of a parametric modeling technique used to automate the quantitative aspects of ISHH. The ISHH experiments described here utilized a specific DNA probe complementary to mRNA molecules encoding the neuropeptide substance P and related tachykinin peptides. A nonlinear model was used to describe the dark-field intensity pattern of labeled neurons. The model's parameters were then employed in detecting individually hybridized neurons and in estimating levels of preprotachykinin mRNA and associated cellular areas. Total mRNA content was quantified by relating the intensities described by the models to those obtained from 14C autoradiographic standards. Finally, the algorithm's performance was evaluated by comparing these estimates to those obtained from manual grain counts of labeled neurons. Overall, the parametric model presented here facilitates the process of performing quantitative analysis of hybridized neurons based on predetermined and unbiased morphological criteria. PMID- 9343567 TI - Semiautomatic image analysis for grain counting in in situ hybridization experiments. AB - We have developed a computer image analysis procedure for counting autoradiographic grains in in situ hybridization experiments. The procedure automatically estimates the number of autoradiographic grains over cells and measures cell number and size so that grain density per unit cell area can be calculated. Advantages include the clear separation of grains and cells, using chromatic and spatial filters to enhance the image; the use of gray level operators to extract cells from grains; and the use of binary operators for separating apposed or partially overlapping cells and grains. Comparison of manual and automated grain counts revealed a significant correlation between human and computer estimations of grain number. However, the automatic grain counting technique consistently underestimated the number of grains when grain density was high. Measures of the fractional area occupied by grains normalized by the average area of a single grain were a better estimate at high grain densities. The procedure can be modified easily to operate on most image analyzers. PMID- 9343568 TI - Rat brain acetylcholinesterase visualized with [11C]physostigmine. AB - Physostigmine, a powerful cholinesterase inhibitor, has recently been labelled with 11C in view of its potential application for in vivo imaging of cerebral acetylcholinesterase (AChE) using positron emission tomography. Here we carried out autoradiography of the rat brain using [11C]physostigmine in order to characterize the cerebral targets of this ligand. Autoradiograms were obtained using phosphor storage plates which, compared to autoradiographic films, greatly improved the quality of 11C images. Following autoradiography, brain sections were stained for AChE activity, allowing a direct comparison of autoradiographic and histoenzymatic localizations. The distributions of 11C label and of AChE activity were found to be essentially super-imposable, both after in vivo injection of and after in vitro incubation with [11C]physostigmine. Densitometric analysis showed that radioactivity and enzymatic activity distributions were regionally correlated. The fixation of [11C]physostigmine to cerebral tissue was abolished after incubation of the rat brain sections with BW 284C51, a specific AChE inhibitor, but not after incubation with iso-OMPA, a specific inhibitor of butyrylcholinesterase. Unilateral excitotoxic lesions of the striatum that eliminated local AChE expression concomitantly reduced the binding of the ligand in the lesioned area. These results indicate that autoradiographic images of the rat brain obtained with [11C]physostigmine reflect AChE distribution, thus supporting the use of this radioligand to trace cerebral AChE activity in humans with positron emission tomography. PMID- 9343569 TI - Imaging optical reflectance in rodent barrel and forelimb sensory cortex. AB - Novel neuroimaging techniques are extending the scope for studying dynamic brain function. We have developed a system which enables the repeatable imaging of rapid function in rodent primary somatosensory cortex (S-I), based on activity related changes in its optical reflectance (intrinsic signals). The S-I cortices of anesthetized male Sprague-Dawley rats were exposed. Images were acquired with a slow-scan, cooled, charge-coupled device camera (CCD) through filters at 550, 610, and 850 nm before, during, and after contralateral stimulation (vibrissal deflection or forepaw stimulation). Images were divided by prestimulus controls and then averaged across 9-27 trials to produce maps of stimulus-related reflectance change. Optical activity had magnitude 10(-3) of baseline reflectance and consistently comprised two distinct spatiotemporal components over cortex, depending on paradigm. The diffuse signal at 610 nm begins 0.5-1 s after stimulus onset and has a duration of 4-5 s. The second signal is macrovenous and is delayed by 1 s. Similar response patterns were observed at 550 and 850 nm. Evoked potentials, recorded at sites inside and outside the zone of optical activity, confirmed the functional nature of these signals. Using a CCD we have imaged functional reflectance changes over rodent S-I which commence, peak, and extinguish over a time scale of seconds. This optical activity is consistent with the etiologies of microvascular recruitment and chromophore redox change. PMID- 9343570 TI - Evaluating and validating two methods for estimating brain structure volumes: tessellation and simple pixel counting. AB - Developments in imaging technology have made three-dimensional visualization of internal brain structures possible with excellent resolution. Since improved visualization implies improved measurement, these advances hold promise to more accurately measure the volumes of internal structures. As new technologies and techniques emerge, evaluating the relative benefits of measurement methods becomes necessary. We compared and evaluated two methods of estimating volumes from images of brain structures. One method counted pixels within a region of interest, while the other method tessellated the surface between tracings on adjacent slices. Our study assessed both measurement error for true phantom volumes and method disparity for in vivo structures in a randomly selected sample of subjects (n = 100). For our comparisons, we focused on the temporal lobe, ventricular system, and hippocampus. Bias, independence of measurement errors and maximal discrimination of individual differences are properties that are relevant to validating and evaluating measurements of cerebral structure. Pixel counting proved to be the more robust of the two methods, being less sensitive to nuisance interactions between size of object, shape, and slice thickness. Clinical and research applications of imaging techniques may have distinctive but overlapping needs when evaluating and validating new developments in imaging. PMID- 9343571 TI - Development of glomerular structure in rabbit olfactory bulb: three-dimensional reconstruction under the confocal laser scanning microscopy. AB - Confocal laser scanning imaging was used to reconstruct the three-dimensional distribution of the aggregates of olfactory receptor axons terminating within individual glomeruli in the rabbit main olfactory bulb. Two monoclonal antibodies, R2D5 and R4B12, were used to mark selectively the olfactory receptor axons. Thick coronal sections (100-200 microns in thickness) through the bulb were labeled with either of the antibodies, and then serial optical sectioning was performed to reconstruct three-dimensional optic images of the labeled axons in the glomeruli. The results revealed an intricate internal structure of the glomeruli with a mesh-like arrangement of bundles of terminal olfactory axons. Developmental study showed that primitive glomeruli lacking the internal structure of the adult form first appear in the 22-day embryo and that the characteristic internal structure of the glomeruli develops mainly during postnatal days. The confocal laser scanning image method together with specific molecular markers provides a simple tool for the three-dimensional analysis of the glomerular structure. PMID- 9343572 TI - Cordance: a new method for assessment of cerebral perfusion and metabolism using quantitative electroencephalography. AB - Increased slow-wave and decreased fast-wave activity on the electroencephalogram is common in brain dysfunction and may be caused by partial cortical deafferentation. No measure that is specific or sensitive for this deafferentation, however, has yet been reported. We studied a series of subjects with white-matter lesions undercutting the cortex and developed a method for analyzing electrical activity called "cordance" that has face validity as a measure of cortical deafferentation. Cordance is measured along a continuum of values: positive values denote "concordance," an indicator associated with normally functioning brain tissue; negative values denote "discordance," an indicator associated with undercutting lesions, low perfusion, and low metabolism. We present a series of subjects studied with magnetic resonance imaging, positron emission tomography, and single-photon emission computed tomography that demonstrate strong associations between cordance and other measures of brain structure and function. PMID- 9343573 TI - Near-microscopic magnetic resonance imaging of the brains of phenylalanine hydroxylase-deficient mice, normal littermates, and of normal BALB/c mice at 9.4 Tesla. AB - The near-microscopic resolution of the mouse brain, by magnetic resonance imaging (MRI) at 9.4 T, permits in situ examination of the entire brain and longitudinal studies of neural development. MRI can be utilized to reveal brain structure at a resolution of 100 microns in the X, Y, and Z planes of brain, to differentiate the gray from white (myelin-rich) matter, and to reveal the ventricular compartments. The present report describes the structure of normal BALB/c mouse brain as revealed by imaging at 9.4 T and by histological stains; the structure of normal brain is compared with that from a phenylalanine hydroxylase-deficient mouse mutant line (Pah(enu2)) and those from normal littermates. The brains of patients with phenylketonuria (PKU) were reported to have demyelination and other structural abnormalities revealed by magnetic resonance imaging (MRI). Therefore, high-resolution MRI was used to examine the brain of this mutant, an animal model for the study of human phenylketonuria. Our study revealed no evidence of demyelination or other abnormalities in the brains of Pah(enu2) mice. Histologically, the mutant and normal mouse brains appear similar. This is consistent with a recent study from our laboratory which demonstrated that the histology of the brain of an untreated male patient, who died with PKU at the age of 29, was similar to control brain with the exception of changes directly related to visual blindness and seizures experienced by the patient. PMID- 9343574 TI - Serial section electron tomography: a method for three-dimensional reconstruction of large structures. AB - We present a method for combining single axis tomography and serial sectioning techniques to derive a three-dimensional reconstruction of large structures at electron microscopic resolution. This serial-tomography method allows the use of sufficiently thin sections to achieve adequate resolution with electron tomography, yet enables the generation of large reconstructions with considerably fewer sections than would be required using a serial thin section reconstruction technique. Serial thick sections (1-2 microns) are cut through the structure of interest, tomographic volume reconstructions are obtained for each section from a single axis tilt series, and the resulting series of volumes are then aligned and combined to form a single large volume. The serial-tomography method is illustrated with several samples, including red blood cells, the Golgi apparatus, and a spiny dendrite of a cortical pyramidal neuron. In some of these samples, the reconstruction is compared to correlated light microscopic views. The resulting large volume reconstructions appear to represent accurately the size and shape of objects such as red blood cells and spiny dendrites. The continuity of complex, tortuous structures such as the Golgi apparatus is also maintained across serial volumes. These examples demonstrate that it is possible to align and link a series of tomographic volumes accurately and that serial-tomography is a useful method for reconstructing relatively large structures without resorting to large numbers of serial thin sections. PMID- 9343575 TI - Fast multisite optical recording of mono- and polysynaptic activity in the hamster suprachiasmatic nucleus evoked by retinohypothalamic tract stimulation. AB - Responses of the hamster suprachiasmatic nucleus (SCN) to retinohypothalamic tract (RHT) stimulation were studied in horizontal hypothalamic slices using fast multisite optical recording techniques. A 124-element photodiode detector array provided high-speed monitoring (0.5 ms/frame) of evoked neural activity in the SCN, while a larger 464-element photodiode array yielded improved spatial imaging with some loss in temporal resolution (1.6 ms/frame). Brief electrical stimulation of the optic nerves evoked a propagated compound action potential that was recorded optically as a single transient depolarization in many slice regions, including the SCN. Only within the SCN, however, was this optic tract signal followed by additional voltage-dependent optical responses which exhibited a fast and a slow depolarizing component. The initial upstroke of the fast component was Ca(2+)-insensitive and is presumed to reflect activity in presynaptic RHT afferents. The remainder of the fast depolarization and the slow depolarization were Ca(2+)-sensitive. These responses were labeled the early population excitatory postsynaptic potential (Early P.E.P.S.P.) and the Late P.E.P.S.P. respectively. The Late P.E.P.S.P. was not enhanced by K+ channel blockade, suggesting that glial depolarization is not the primary source of this component. Drugs known to suppress RHT-evoked SCN field potentials also suppressed the Early and Late P.E.P.S.P.'s recorded optically in the SCN. Unexpectedly, the Early P.E.P.S.P. was also reduced by the GABAA antagonist, bicuculline. Surface plots of normalized peak amplitudes showed that both SCN components had similar spatial distributions within the SCN, although the Early P.E.P.S.P. tended to be slightly more prominent within the medial SCN in some preparations. It is suggested that the Early P.E.P.S.P. represents firing of monosynaptically activated SCN neurons, while the Late P.E.P.S.P. reflects polysynaptic activity within the intrinsic SCN neuronal network that may be involved in the light entrainment of the circadian oscillator. PMID- 9343576 TI - Incorporation of fluorescent lipids into living rabbit hippocampal and cerebellar slices. AB - Incorporation of exogenously applied fluorescent lipids into living cells was exploited to probe cellular structure and function in living hippocampal and cerebellar slices as assessed by fluorescent imaging techniques and intracellular recording. Nitrobenzoxadiole-phosphatidylcholine (NBD-PC) and BODIPY phorbol ester, in vitro substrates of phospholipase activity and protein kinase C, respectively, were incorporated and distributed into specific cell populations. In the hippocampal slice, both probes labeled the somata and proximal dendrites of pyramidal and granule cells but were hetrogeneously distributed across the different hippocampal fields. Changes in fluorescent properties of NBD-PC in individual pyramidal cell and granule cell somata were quantified upon challenge with a muscarinic agonist known to modulate phospholipase A2 activity. In the cerebellar slice, both probes labeled Purkinje cell bodies and dendrites but only NBD-PC labeled stellate and granule cells. The cellular and functional specificity of these fluorescent lipid probes shows great promise for monitoring biochemical events in complex neuronal systems with significant spatial and temporal resolution. PMID- 9343578 TI - Double-label immunofluorescence with the laser scanning confocal microscope using cyanine dyes. AB - The laser scanning confocal microscope, when used with the krypton-argon ion laser, is well suited for the simultaneous detection of pairs of antigens by immunofluorescence. Traditionally, double-label studies have utilized secondary antibodies conjugated to fluorescein isothiocyanate (FITC), excited by the 488-nm line (blue), and to tetramethyl rhodamine isothiocyanate or Texas Red, excited by the 568-nm line (yellow). However, the use of fluorophores excited by the 488 nm line produces unsatisfactory results when tissue contains low wavelength excitable autofluorescence. In the amphibian cardiac ganglion, for example, autofluorescent granules within parasympathetic neurons obscure cell surface derived signals and prevent one from analyzing the relative position of acetylcholine receptor clusters and synaptic boutons by double-label immunofluorescence. This problem has been solved by using cyanine 3.18 (Cy3)- and cyanine 5.18 (Cy5)-conjugated secondary antibodies, which are excited efficiently by the 568-nm (yellow) and the 647-nm (red) lines and which emit in the orange/red and in the far-red, respectively, and thus by avoiding the 488-nm line altogether. The resulting images are as good or better than those obtained with FITC and Texas Red, even without consideration of autofluorescence. PMID- 9343577 TI - Identification of acutely isolated cells from developing rat cerebellum. AB - Immunocytochemical staining was used to identify nerve and glial cells from postnatal rat cerebelli in situ and following tissue dissociation. Purkinje cells were identified using antibodies for the calcium-binding proteins calbindin and PEP19. Purkinje cells isolated during the second postnatal week were 15-20 microns in diameter and relatively abundant and displayed thin perisomatic processes. These features were used to identify Purkinje cells with scanning electron microscopy, which revealed extensive membrane infoldings. Golgi and nuclear cells were identified using antibodies against rat-303 antigen. Pale, nuclear, and Purkinje cells were identified using antibodies for rat-302 antigen. Although staining for rat-302 and rat-303 was weak during the second postnatal week, we were able to identify Golgi and pale cells even after tissue dissociation. Isolated Golgi cells were 8-10 microns in diameter and fewer in number than Purkinje cells and did not counterstain with calbindin antibodies. Isolated pale cells were 8-10 microns in diameter, rare, and resistant to calbindin antibodies. Isolated neurons from cerebellar nuclei were not located with either 302 or 303 staining, suggesting that they remained in the tissue. Golgi-Bergmann cells and astrocytes were identified using antibodies for glial fibrillary acidic protein. Isolated glial cells were 12-15 microns in diameter, more numerous than Purkinje cells, and unstained with calbindin antibodies. With phase-contrast optics, glial cells appeared flatter than neuronal cell types and had acentric nuclei. These results demonstrate that specific cell types in developing rat cerebellum can be identified after acute isolation, which should facilitate analysis of their endogenous properties. PMID- 9343579 TI - Three-dimensional reconstructions of the developing forebrain in rat embryos. AB - Using a computerized three-dimensional reconstruction technique with serially sectioned rat embryos, changes in the size and form of the forebrain were studied on Embryonic Days (E) 12 (1 day after closure of the neural tube), E15, E18, and E21 (2 days before birth). During this time, the forebrain changes from a relatively simple tubular structure with thin walls surrounding a large ventricular system to a thick-walled brain with a highly convoluted but reduced ventricular system. On E12, the two components of the forebrain, the telencephalon and the diencephalon, cannot be distinguished. Considering the forebrain as a whole (the embryonic prosencephalon), its volume continually increases between E12 and E21 due to the generation, differentiation, and maturation of neurons and glia. Attention was paid to changes in the sizes of the ventricles, the neuroepithelium and the parenchyma. Volumes of the ventricles and the surrounding neuroepithelium rapidly expanded from E12 to E18 and then decreased by E21, while the volume of the parenchyma continually increased. Differential growth of the telencephalon and that of the diencephalon were compared between E15 and E21. The expansion of the telencephalon was much larger than that of the diencephalon. In the telencephalon, the volumes of the lateral ventricles and the surrounding neuroepithelium increased between E15 and E18 and decreased by E21, while in the diencephalon the volumes of the third ventricle and its surrounding neuroepithelium continually declined between E15 and E21. That observation is compatible with previous work showing that the majority of diencephalic structures develop earlier than those in the telencephalon. It is important to note that volume changes in the ventricles and the neuroepithelium are maintained in "lock-step," suggesting a close relationship between the size of the ventricle and the size of the neuroepithelium. PMID- 9343580 TI - Hyperbaric oxygen treatment in acute experimental allergic encephalomyelitis. Contribution of magnetic resonance imaging study. AB - Magnetic resonance imaging (MRI) has been performed to determine the efficacy of hyperbaric oxygen treatment (HBO) on experimental allergic encephalomyelitis (EAE) in Lewis rats. The animals were exposed for 2 x 3 h in a 24-h period at 2 ATA pure O2. Three HBO treatment protocols were used: a 10-day preventive treatment (beginning on the 1st day postimmunization), a 3-day preventive treatment (beginning on the 1st day postimmunization), and a 10-day symptomatic treatment (beginning on the 11th day postimmunization). Based on clinical and MRI observations, this study demonstrates: (i) that HBO treatment does not reduce the blood-brain barrier (BBB) disturbance and cerebral edema in EAE, (ii) that in an opposite way, it provokes reversible BBB breakdown, and (iii) that preventive HBO treatments results in modification of the course of EAE, possibly by immunosuppression effect during the initial sensitization step. PMID- 9343581 TI - Evidence for uptake of vital dye by activated rat peritoneal mast cells: an in vitro imaging study. AB - Uptake of material from surrounding medium by activated rat peritoneal mast cells (PMCs) was studied using in vitro peritoneal eluate cells, the vital fluorescent dye sulforhodamine B (SFRM-B), secretagogue compound 48/80, and an imaging technique. PMCs, which undergo different states of degranulation, are shown to possess the ability to take up (by endocytosis) SFRM-B in an activity-dependent manner. The endocytosed dye is incorporated in the granules and can be discharged into the medium when the cells are reactivated. Both the uptake and the discharge processes are calcium-dependent. The reactivity of mast cells to secretagogue is not altered by the application of the dye. SFRM-B, a negatively charged, nonspecific protein stain, displays greater photostability and less leakage than the positively charged acridine orange, and its fluorescence persists for hours, whereas acridine orange fluorescence fades within 1 min when exposed to ultraviolet illumination. The fluorescent image of the dye-loaded mast cells can be preserved overnight in a container at room temperature. SFRM-B elicits no detectable damaging influence on the activated afferent discharge of splanchnic afferent nerve fibers with mesenteric terminals. This enables the use of SFRM-B for studying the interactions between mesenteric afferent terminals and their surrounding mast cells. PMID- 9343582 TI - Application of the Cavalieri principle in volume estimation using laser confocal microscopy. AB - The Cavalieri principle, a well-established stereological technique, uses interpolation between samples to estimate volume of three-dimensional (3D) objects. Serial optical sectioning with the confocal microscope resembles certain aspects of the Cavalieri principle, albeit with no interpolation. However, reconstruction and analysis of finely spaced optical sections can be cumbersome and time consuming. Application of the Cavalieri principle to confocal sections may be advantageous in reducing the size of the data set required to obtain reliable estimates of volume. In the present study, somal volumes of phrenic motoneurons were estimated by applying the Cavalieri principle to confocal images. These estimates were compared to measurements of somal volume using on interpolation of confocal sections. Phrenic motoneurons in adult rats were retrogradely labeled with a fluorescent rhodamine dye. Confocal optical sections of 0.6 micron thickness were then obtained from 150-micron-thick spinal cord slices containing labeled neurons. These image sets were reoriented to represent transverse sections. The Cavalieri principle was applied to these confocal image sets at selected sampling intervals from 1.2 to 3.0 microns. Planimetric measurements of motoneuron somal cross-sectional area in the selected sections were made using a point-counting method. At sampling intervals less than 2.4 microns, individuals motoneuron somal volume estimates were similar for the noninterpolated confocal and the interpolated Cavalieri methods. At these sampling intervals, the distributions of motoneuron somal volumes were also similar for the two methods. At a sampling interval of 2.4 microns or greater, there was a greater variability in individual motoneuron somal volume estimates, although the population mean and median were similar to the noninterpolated confocal measurements. Therefore, a satisfactory agreement between noninterpolated confocal measurements and the Cavalieri estimates suggests that less-stringent optical sectioning parameters may suffice for individual cell volume measurements when using confocal microscopy, thus making it significantly more efficient. PMID- 9343583 TI - High-resolution anatomy from in situ human brain. AB - We have generated a spatially accurate, high-resolution three-dimensional (3D) volume of brain anatomy from cryosectioned whole human head. The head of a female cadaver was cryosectioned on a heavy duty cryomacrotome (PMV, Stockholm Sweden) modified for quantitative digital image capture. Serial images (1024(2), 24-bit) were captured directly from the cryoplaned specimen blockface in 500-micron intervals and reconstructed to a 3D data volume. Data were placed into the Talairach coordinate system to create a volume of brain anatomy for atlas reference. We resampled the volume at 500 microns along the sagittal, coronal, and horizontal planes and enhanced the images by digitally editing the background. The spatial resolution of the original digitized images provided sufficient anatomic detail to clearly delineate gray and white matter and neural structures, including major fiber pathways, subthalamic nuclei, and laminae. We developed a compact disk and controlling software program to enable the viewer to select planes of orientation, display, and copy individual to sections at higher resolution. Animation proved useful in the conveyance of system anatomy as structures are shown traversing through the neuroaxis. Postmortem cryosectioning paired with this computerized presentation allowed the complete 3D volume data to be distributed and shared as an educational, clinical, and research resource. PMID- 9343584 TI - Mouse lemur microscopic MRI brain atlas. AB - We present a three-dimensional (3D) digital atlas of a mouse lemur head at 60 micron cubic voxel isotropic resolution. It was constructed from a 3D proton magnetic resonance image (MRI) acquired at 500 MHz. It shows views of 3D volume rendering and movies of contiguous 2D plane cuts in three orthogonal directions. This MR data set was acquired using a spin-echo pulse sequence under conditions of strong T2 and significant diffusion weighting. Experimental parameters were optimized to provide strong intrinsic contrast between gray and white matter. Familiar anatomical structures are clearly identifiable. Fine fiber tracts, laminations of cortices, details of the inner ears, and layering in the lateral geniculate nuclei are all visible. Anatomical identifications of structures in representative slices selected from the atlas are presented. We also describe some difficulties and trade-offs encountered in microscopic resolution MRI. We note that this type of atlas does not suffer from the spatial distortions and slice registration problems introduced by standard histological techniques. Future in vivo longitudinal studies will provide atlases describing in detail the development of the primate brain. PMID- 9343585 TI - Dynamic imaging of purified individual synaptic vesicles. AB - The atomic force microscope (AFM) was used to directly image purified synaptic vesicles. Individual secretory vesicles (approximately 50 nm diameter) were resolved with the AFM when imaged either dry or in solution. Vesicles were observed repeatedly for periods of greater than 2 h. To ask whether the AFM can detect structural change of vesicles the osmolarity of the bathing medium was reduced from 330 to 110 mOsm. Hypo-osmotic treatment caused an expansion and flattening of the vesicles. Thus, using the AFM it is possible to resolve individual vesicles and follow changes in vesicular structure. This opens the possibility that the secretory event can be reconstituted and visualized in vitro in order to elucidate the roles of synaptic proteins in synaptic transmission. PMID- 9343586 TI - Three-dimensional reconstruction of activated columns from 2-[14C]deoxy-D-glucose data. AB - Three-dimensional (3-D) reconstruction of autoradiograms can provide new insights into the functional relationship of neural regions. To reach full potential, however, 3-D reconstruction must be both accurate and efficient. In this paper, we present a novel image matching algorithm that simultaneously aligns a set of serial sections and uses the method to reconstruct whisker barrels from the rat cerebral cortex. We initially compared several alignment techniques and found that our Multi-Set Registration (MSR) algorithm produced superior accuracy. This algorithm is based on a least-squares minimization technique and is able to simultaneously register a set of serial sections with subpixel precision (30 micron accuracy). We applied our new technique to the 3-D reconstruction of a series of autoradiograms. Our objective was to visualize and measure the 3-D metabolic (functional) shape of normal (control) and developmentally altered (plastic) C3 vibrissa columns in the first somatosensory area of the rat cerebral cortex. The plastic C3 metabolic column showed a nearly 450% increase in volume when compared to the control column. In addition, the lesion-altered C3 column-in contrast to the normal C3 column-displayed no central zone of high activity, and patches of higher metabolic activity were scattered throughout the columnar profile. This metabolic activity was not confined to the cylindrical column, but extended tangentially as radiating fingerlike projections toward neighboring barrels. PMID- 9343587 TI - Three-dimensional reconstruction of the rubrocerebellar premotor network of the turtle. AB - Neuroanatomical studies have demonstrated that the organization of the reptilian rubrocerebellar limb premotor network is similar to that of mammals. This network is composed of prominent recurrent connections among the red nucleus, lateral cerebellar nucleus and lateral reticular nucleus. In this paper the rubrocerebellar system of the turtle was three-dimensionally reconstructed to permit detailed examination of its anatomical organization. Each nucleus and its major efferent pathway was imaged and reconstructed from separate anatomical cases. Section images were used to draw tissue boundaries, mark cell positions and locate axonal trajectories. For each nucleus, drawings of section images containing labeled cells were stacked in the rostrocaudal direction using anatomical landmarks, and a graphic model of the surface was constructed using the method of triangulation. An ellipsoid of equal concentration was computed for each nucleus to ascertain their three-dimensional boundaries and location within the brainstem. To examine the entire rubrocerebellar network, a template of the turtle brainstem and cerebellum was constructed. The component nuclei of the rubrocerebellar network and their axonal projections were then spatially warped onto the template reconstruction on a section by section basis. The final three dimensional reconstruction of the turtle rubrocerebellar limb premotor network could be rotated in space, allowing proper visualization of the anatomical details of this system. Furthermore, we were able to mathematically section through the reconstruction to obtain brainstem slices with differing orientations and thickness. PMID- 9343588 TI - A landscape parametric profile approach for rat brain image analysis. AB - This paper develops the use of the landscape parametric profile as a descriptive and interpretative method of analyzing metabolic activity data obtained by neuroimaging techniques such as 2-deoxyglucose autoradiography. The method is suggested as an initial survey-type approach that can be used to map regional activity throughout the coronal planes of a rat brain and to compare relative differences in the patterns of activity between different brains. Images are acquired using preexisting image analysis software and then transformed to landscape parametric profiles derived from each matrix array. The image profile analysis tasks are done using commercially available software for matrix computations such as MATLAB. An effective way of interpreting activity differences between brains is presented by plotting the results from a randomized blocks ANOVA significance test combined with the average profiles of different subjects within the same experimental group. The method greatly reduces the computational burden associated with image averaging and statistical comparison of brains from different subjects while preserving the richness in topographic metabolic information. One significant advantage of the method is that it results in figures that are readily depicted and interpretable in black and white. PMID- 9343589 TI - Analysis of fMRI time-series revisited. AB - This paper presents a general approach to the analysis of functional MRI time series from one or more subjects. The approach is predicated on an extension of the general linear model that allows for correlations between error terms due to physiological noise or correlations that ensue after temporal smoothing. This extension uses the effective degrees of freedom associated with the error term. The effective degrees of freedom are a simple function of the number of scans and the temporal auto correlation function. A specific form for the latter can be assumed if the data are smoothed, in time, to accentuate hemodynamic responses with a neural basis. This assumption leads to an expedient implementation of a flexible statistical framework. The importance of this small extension is that, in contradistinction to our previous approach, any parametric statistical analysis can be implemented. We demonstrate this point using a multiple regression analysis that tests for effects of interest (activations due to word generation), while taking explicit account of some obvious confounds. PMID- 9343590 TI - Autoradiographic evidence that QNB displays in vivo selectivity for the m2 subtype. AB - Alzheimer's disease (AD) involves selective loss of muscarinic m2, but not m1, subtype neuroreceptors in cortical and hippocampal regions of the human brain. Emission tomographic study of the loss of m2 receptors in AD is limited by the fact that there is currently no available m2-selective radioligand which can penetrate the blood-brain barrier. We have previously reported the results of in vivo dissection studies, using both carrier-free and low specific activity [3H]QNB, which show that [3H]QNB exhibits a substantial in vivo m2 selectivity. Because of the expense of the radioligand and the long exposure time required for the X-ray film, performing a large number of direct in vivo autoradiographic studies using [3H]QNB is precluded. Therefore, we now confirm these results autoradiographically by studying the in vivo inhibition of radio-iodinated (R)-3 quinuclidinyl (S)-4-iodobenzilate ((R,S)-[125I]IQNB) binding by unlabeled QNB. In the absence of QNB, (R,S)-[125I]IQNB labels brain regions in proportion to the total muscarinic receptor concentration; in the presence of 15 nmol QNB, (R,S,) [125I]IQNB labeling in those brain regions containing predominantly m2 subtype is reduced to background levels. We conclude that QNB is m2-selective in vivo and that a suitably radiolabeled derivative of QNB, possibly labeled with 18F, may be of potential use in positron emission tomographic study of the loss of m2 receptors in AD. PMID- 9343591 TI - Novel method for stereo imaging in light microscopy at high magnifications. PMID- 9343592 TI - A probabilistic atlas of the human brain: theory and rationale for its development. The International Consortium for Brain Mapping (ICBM). PMID- 9343593 TI - The innervation of human skin studied with confocal scanning laser microscopy: a comparison between PGP 9.5 immunofluorescence and silver impregnations. PMID- 9343594 TI - Brain activation induced by the perceptual maze test: a PET study of cognitive performance. AB - We investigated with PET the cerebral activation pattern elicited by the perceptual maze test (PMT), a neuropsychological test used to evaluate organic brain injury. The PMT examines visuospatial skill, general intelligence, visually guided motor planning, and the ability to obey rules. Eight right-handed volunteers were examined with PET using the tracer [15O]butanol. Three paradigms containing the PMT, a motor control (SHAM), and a rest condition were examined twice in a randomized order. Solving the PMT caused extensive bilateral activations in the occipital lobe extending rostrally into the parietal lobe and caudally to the posterior part of the temporal lobe. Bilateral activations were also seen in the prefrontal, medial premotor, and the anterior cingulate cortex (ACC). The premotor and primary sensory motor cortices contralateral to the performing hand were also activated. Marked activations were noted in the visual system, including areas pertaining to visuospatial decoding. The previously defined functional network (ACC, prefrontal and posterior parietal cortex) for the maintenance of visuospatial attention was activated during the PMT. Extensive bilateral deactivations were seen in frontomedial, temporal, parietal, and posterior cingulate regions. This pattern may represent relatively decreased blood flow in cortical areas pertaining to sensory modalities that were not activated in the PMT. The decreased activity in these regions could also express diminished cognitive processing in neuronal systems that might interfere with the task-related performance. PMID- 9343595 TI - Retinotopic maps in human prestriate visual cortex: the demarcation of areas V2 and V3. AB - We have used PET (positron emission tomography) to chart the mapping of the retina in human occipital visual cortex and hence to locate the secondary and tertiary visual areas, V2 and V3. A group of four non-selected male volunteers was presented with dynamic stimuli that were aligned with either the vertical or the right horizontal meridians (VM or HM) from 0 degree to 29 degrees eccentricity; the vertical stimuli were restricted to either the inferior or the superior hemifields. PET scans were performed using intravenous infusion of H215O and a Siemens-CTI 953B PET scanner with 3D data acquisition. Subjects received 18 scans, divided equally among the right HM, the superior VM, and the inferior VM. Data were analyzed with SPM software. The group average result confirmed our experimental hypothesis that human occipital visual cortex has retinotopic maps similar to those of the macaque monkey. Thus human areas V2 and V3 can be defined on the basis that the border between them is formed by the HM and that the outer border of V3 is demarcated by a second representation of the VM that runs approximately parallel to the primary representation of the VM at the V1/V2 border. Furthermore, as in many mammals, the extrastriate representation of the HM is "split", such that the superior contralateral quadrant is mapped in lower V2 and V3, occupying the ventral surface of human cortex, and the inferior contralateral quadrant is mapped in upper V2 and V3, which extend over the lateral and medial surfaces of each hemisphere. After stereotaxic normalization, the position of V3 defined by retinal topography was found to correspond to that surmised from our previous PET studies employing moving stimuli. PMID- 9343596 TI - Computerized brain tissue classification of magnetic resonance images: a new approach to the problem of partial volume artifact. AB - Due to the finite spatial resolution of digital magnetic resonance images of the brain, and the complexity of anatomical interfaces between brain regions of different tissue type, it is inevitable that some voxels will represent a mixture of two or three different tissue types. Outright assignment of such "bipartial" or "tripartial" voxels to one class or another is more problematic and less reliable than assignment of "full-volume" voxels, wholly representative of a single tissue type. We have developed a computerized system for brain tissue classification of dual echo MR data, which uses a polychotomous logistic model for discriminant analysis, combined with a Bayes allocation rule incorporating differential prior probabilities, and spatial connectivity tests, to assign each voxel in the image to one of four possible classes: gray matter, white matter, cerebrospinal fluid, or unclassified. The system supports automated volumetric analysis of segmented images, has low operational overheads, and compares favorably with previous multivariate or "multispectral" approaches to brain MR image segmentation in terms of both validity (bootstrap misclassification rate = 3.3%) and interoperator reliability (intra-class correlation coefficients for all three tissue classes > 0.9). We argue that these improvements in performance stem from better methodological management of the related problems of non-Normality of MR signal intensity values and partial volume artifact. PMID- 9343597 TI - Review: does measurement of regional cerebral blood flow reflect synaptic activity? Implications for PET and fMRI. AB - The energy metabolism of the adult human brain almost completely depends on glucose. The functional coupling of regional cerebral blood flow and local cerebral glucose metabolism has been established in a wide range of experiments using autoradiographic techniques in rats, cats, and monkeys as well as double tracer techniques in humans. Glucose utilization in turn reflects neuronal activity and more specifically synaptic, mainly presynaptic, activity. The majority of glucose is needed for the maintenance of membrane potentials and restoration of ion gradients. PET as well as fMRI may be used to study changes in blood flow or flow-related phenomena in human subjects in vivo. Both techniques monitor changes of synaptic activity in a population of cells. These changes may be due to excitation or inhibition. More than 85% of cerebral glucose is used by neurons (mainly presynaptic axon terminals), while the remainder may at least partly account for metabolic processes in glial cells. Monitoring of regional cerebral blood flow with PET or fMRI thus mainly reflects neuronal and more specifically (pre-) synaptic activity. PMID- 9343598 TI - Characterizing evoked hemodynamics with fMRI. AB - We describe an implementation of the general linear model that facilitates the characterization of evoked hemodynamic responses to sensorimotor or cognitive processing, when the exact form of these responses is not known. The importance of this approach is that one can test for differential responses among tasks that may elude more conventional analyses. In particular, we suppose that an evoked response has early and late components and that a differential response may involve (i) both components to the same degree, as in a conventional "activation" or (ii) differential expression of the early and late components in two tasks, as might be seen in differential adaptation, or differences associated with the tasks (e.g., requiring and not requiring sustained attention). Using this approach we were able to demonstrate that the anterior cingulate differentiates, in terms of its response, between two motor tasks that did and did not require sustained attention. This differential response was observed even though there was no classical "activation" (i.e., there was no difference in the mean activity associated with the two conditions). It is suggested that these demonstration results point to the possibility of making greater use of the temporal resolution afforded by fast fMRI techniques. PMID- 9343599 TI - Characterizing dynamic brain responses with fMRI: a multivariate approach. AB - In this paper we present a multivariate analysis of evoked hemodynamic responses and their spatiotemporal dynamics as measured with fast fMRI. This analysis uses standard multivariate statistics (MANCOVA) and the general linear model to make inferences about effects of interest and canonical variates analysis (CVA) to describe the important features of these effects. We have used these techniques to characterize the form of hemodynamic transients that are evoked during a cognitive or sensorimotor task. In particular we do not assume that the neural or hemodynamic response reaches some "steady state" but acknowledge that these physiological changes could show profound task-dependent adaptation and time dependent changes during the task. To address this issue we have modeled hemodynamic responses using appropriate temporal basis functions and estimated their exact form within the general linear model using MANCOVA. We do not propose that this analysis is a particularly powerful way to make inferences about functional specialization (or more generally functional anatomy) because it only provides statistical inferences about the distributed (whole brain) responses evoked by different conditions. However, its application to characterizing the temporal aspects of evoked hemodynamic responses reveals some compelling and somewhat unexpected perspectives on transient but stereotyped responses to changes in cognitive or sensorimotor processing. The most remarkable observation is that these responses can be biphasic and show profound differences in their form depending on the extant task or condition. Furthermore these differences can be seen in the absence of changes in mean signal. PMID- 9343600 TI - Analysis of fMRI time-series revisited--again. AB - Friston et al. (1995, NeuroImage 2:45-53) presented a method for detecting activations in fMRI time-series based on the general linear model and a heuristic analysis of the effective degrees of freedom. In this communication we present corrected results that replace those of the previous paper and solve the same problem without recourse to heuristic arguments. Specifically we introduce a proper and unbiased estimator for the error terms and provide a more generally correct expression for the effective degrees of freedom. The previous estimates of error variance were biased and, in some instances, could have led to a 10-20% overestimate of Z values. Although the previous results are almost correct for the random regressors chosen for validation, the present theoretical results are exact for any covariate or waveform. We comment on some aspects of experimental design and data analysis, in the light of the theoretical framework discussed here. PMID- 9343601 TI - Tests for distributed, nonfocal brain activations. AB - Most approaches to detecting changes in functional brain images assume that activations are focal or very localized. However, the brain's response to cognitive of sensorimotor challenge may be spatially or anatomically distributed. In this paper we consider the usefulness of a test based on the mean sum of squares of statistical parametric maps. The performance of this test is evaluated using simulated and real data and is compared to the gamma 2 test, a test of the size of the activated region, and a focal activation test based on the intensity of local maxima. We demonstrate that the mean sum of squares test is more sensitive to nonfocal signals and propose that it could be used to complement approaches that are more sensitive to focal activations. PMID- 9343602 TI - The mind's eye--precuneus activation in memory-related imagery. AB - We examined brain activity associated with visual imagery at episodic memory retrieval using positron emission tomography (PET). Twelve measurements of regional cerebral blood flow (rCBF) were taken in six right-handed, healthy, male volunteers. During six measurements, they were engaged in the cued recall of imageable verbal paired associates. During the other six measurements, they recalled nonimageable paired associates. Memory performance was equalized across all word lists. The subjects' use of an increased degree of visual imagery during the recall of imageable paired associates was confirmed using subjective rating scales after each scan. Memory-related imagery was associated with significant activation of a medial parietal area, the precuneus. This finding confirms a previously stated hypothesis about the precuneus and provides strong evidence that it is a key part of the neural substate of visual imagery occurring in conscious memory recall. PMID- 9343603 TI - Confocal laser scanning microscopy and 3-D reconstructions of neuronal structures in human brain cortex. AB - Human brain material was studied with Lucifer yellow (LY) microinjections, indirect Texas red immunofluorescence, and confocal laser scanning microscopy (CLSM). The scanned images were transferred to a Silicon Graphics (SG) IRIS computer equipped with software for reconstructing the 3-D architecture of cells. By employing dual channel CLSM (Bio-Rad MRC 600), LY-injected cells and Texas red immunofluorescence could be studied simultaneously. Autopsy material with 2- to 48-h postmortem delays (6 control and 2 Rett's syndrome cases) as well as biopsy material (14 cases with therapy-resistant partial epilepsy--TRPE--undergoing neurosurgery) were used. In each specimen, 100-200 pyramidal and nonpyramidal neurons were visualized by LY microinjection. Single neurons were imaged and 2-D reconstructions of each neuron were made using z-projections of serial optical images; 3-D reconstructions and rotations were computed using the SG workstation, with VoxelView software from Vital Images (UK), and stored in a "neuronal library" on laser or magnetic optical disks. In Ret's syndrome cases and in patients with TRPE various abnormalities in the dendritic geometry of pyramidal and nonpyramidal cells have been found. The combination of LY injections with immunofluorescence allows the investigation of transmitter-related substances around the LY-injected cells. Using antibodies to synaptic vesicle proteins, presynaptic elements docking onto individual spines have been demonstrated. This approach may contribute to the understanding of different neurological and psychiatric disorders and may be useful in the Mapping of the Human Brain project. It may also be integrated with functional imaging by PET scan and with the human genome project. PMID- 9343604 TI - Autoradiographic evidence that quinuclidinyl 4-(bromophenyl)-2-thienylglycolate (QBPTG) displays in vivo selectivity for the muscarinic m2 subtype. AB - Alzheimer's disease (AD) involves selective loss of muscarinic m2, but not m1, subtype neuroreceptors in cortical and hippocampal regions of the human brain. Emission tomographic study of the loss of m2 receptors in AD is limited by the fact that there is currently no available m2-selective radioligand which can penetrate the blood-brain barrier. We now demonstrate the in vivo m2 selectivity of an analogue of QNB, 4-(bromophenyl)-2-thienylglycolate (QBPTG), by studying autoradiographically the in vivo inhibition of radioiodinated (R)-3-quinuclidinyl (S)-4-iodobenzilate ((R,S)-[125I]IQNB) binding by unlabeled QBPTG in rat brain. In the absence of QBPTG, (R,S)-[125I]IQNB labels brain regions in proportion to the total muscarinic receptor concentration; in the presence of 37.5 nmol of racemic QBPTG, (R,S)-[125I]IQNB labeling in those brain regions containing predominantly the m2 subtype is reduced to background levels. We conclude that QBPTG is m2-selective in vivo and that [76Br]QBPTG, or a radiofluorinated analogue, may be of potential use in positron emission tomographic study of the loss of m2 receptors in AD. In addition, a radioiodinated analogue may be of potential use in single photon emission tomographic studies. PMID- 9343605 TI - Activation of association auditory cortex demonstrated with functional MRI. AB - Activations in the temporal lobes previously observed using positron emission tomography and auditory stimuli were partially reproduced with functional MRI and echo-planar imaging at 1.5 T in six volunteers performing tone and phoneme monitoring tasks. Verbal processing compared to a tone recognition task significantly activated a cortical area located in the left anterior temporal region (P < 0.02). PMID- 9343606 TI - Activation of prefrontal cortex in children during a nonspatial working memory task with functional MRI. AB - Functional magnetic resonance imaging (fMRI) was used to examine the pattern of activity of prefrontal cortex in prepubertal children during performance of a nonspatial working memory task. The children observed sequences of letters and responded whenever a letter repeated with exactly one nonidentical letter intervening. In a comparison task, subjects monitored similar sequences of letters for any occurrence of a single, prespecified target letter. Location of activation closely approximated that observed in a recent fMRI study with adults using exactly the same task. Activation of the inferior and middle frontal gyri was reliably observed within individual subjects during performance of the working memory task relative to the comparison task. Activation increased and decreased with a time course that was highly consistent with the task manipulations and correlated with behavioral performance. To our knowledge, this study is one of the first to demonstrate the applicability of fMRI to a normative developmental population. Issues of age dependence of the hemodynamic responses of fMRI are discussed. PMID- 9343607 TI - The motion vision of the blind. PMID- 9343608 TI - Functional segregation of movement-related rhythmic activity in the human brain. AB - Multiple synaptic interconnections in the human brain support concerted rhythmic activity of a large number of cortical neurons, typically close to 10 and 20 Hz. Our present neuromagnetic data provide evidence for distinct functional roles of these spectral components in the somatomotor cortex. The sites of suppression during movement and the subsequent rebound of the 20-Hz rhythm followed, along the motor cortex, the representation of fingers, toes, and mouth, as opposed to the stable origin of the 10-Hz rhythms close to the hand somatosensory cortex. The 20-Hz activity appears to be a signature of active immobilization following movement, whereas the reactive 10-Hz signals likely reflect lack of relevant sensory input from the important upper limbs. PMID- 9343609 TI - A voxel-based method for the statistical analysis of gray and white matter density applied to schizophrenia. AB - We describe a novel technique for characterizing regional cerebral gray and white matter differences in structural magnetic resonance images by the application of methods derived from functional imaging. The technique involves automatic scalp editing of images followed by segmentation, smoothing, and spatial normalization to a symmetrical template brain in stereotactic Talairach space. The basic idea is (i) to convert structural magnetic resonance image data into spatially normalized images of gray (or white) matter density, effected by segmenting the images and smoothing, and then (ii) to use Statistical Parametric Mapping to make inferences about the relationship between gray (or white) matter density and symptoms (or other pathophysiological measures) in a regionally specific fashion. Because the whole brain sum of gray (or white) matter indices is treated as a confound, the analysis reduces to a characterization of relative gray (or white) matter density on a voxel by voxel basis. We suggest that this is a powerful approach to voxel-based statistical anatomy. Using the technique, we constructed maps of the regional cerebral gray and white matter density correlates of syndrome scores (distinct psychotic symptoms) in a group of 15 schizophrenic patients. There was a negative correlation between the score for the reality distortion syndrome and regional gray matter density in the left superior temporal lobe (P = 0.01) and regional white matter density in the corpus callosum (P < 0.001). These abnormalities may be associated with functional changes predisposing to auditory hallucinations and delusions. This method permits the detection of structural differences within the entire brain (as opposed to selected regions of interest) and may be of value in the investigation of structural gray and white matter abnormalities in a variety of brain diseases. PMID- 9343610 TI - Intersubject variability in functional neuroanatomy of silent verb generation: assessment by a new activation detection algorithm based on amplitude and size information. AB - We present an experimental evaluation of a new algorithm for the detection of activated areas in brain functional maps. The new algorithm, named HMSD, is based on a hierarchical multiscale description of the difference image in terms of connected objects. Size and magnitude of each object are simultaneously tested with respect to a bidimensional frequency distribution derived using Monte-Carlo simulations under the null hypothesis. In the present work. HMSD was applied to the analysis of a silent verb generation PET activation protocol conducted in six right-handed subjects. Applied to single-subject data. HMSD reveals activation located in the left inferior frontal gyrus in three subjects (two in the pars opercularis, one in the pars triangularis), and in the pars opercularis of the right inferior frontal gyrus in one case, the latter being combined to a crossed cerebellar activation. Overall, single-case results were consistent with the analyses of stereotactically averaged data. Despite a 2D implentation. HMSD detection performances of averaged data were better than that obtained with the 2D version of statistical parametric mapping (SPM) and comparable to that of the 3D version of SPM. PMID- 9343611 TI - Interference and facilitation effects during selective attention: an H215O PET study of Stroop task performance. AB - To investigate the functional anatomy of interference and facilitation during selective attention, we studied 15 normal subjects using the H215O positron emission tomography technique and a computer presented single-trial Stroop task for cognitive activation. Increases in regional cerebral blood flow (rCBF) were observed in a network of structures that have been previously associated with selective attention, including the anterior cingulate gyrus, the frontal polar cortex, the inferior parietal lobule, and the thalamus, as well as the lingual gyrus. Furthermore rCBF decreases (compared to control states) were observed in lateral extra-striate cortex. rCBF changes in prefrontal and extra-striate regions varied with differences in the need to modulate the influence of word and color information while subjects responded to either incongruent or congruent Stroop stimuli. These results indicate the utility of Stroop procedures for investigating the functional anatomy of selective attention. Given recent interest regarding the role of the anterior cingulate gyrus in the pathophysiology of neuropsychiatric disorders, our results also suggest that the Stroop task can serve as a reliable neurobehavioral probe for this region. The significance of these results for understanding processing mechanisms underlying selective attention is discussed within the framework of a parallel distributed processing model of Stroop task performance. PMID- 9343612 TI - Parametric analysis of functional neuroimages: application to a variable-rate motor task. AB - Positron emission tomography (PET) has proven to be a powerful tool in identifying the functional neuroanatomy underlying cognitive and sensorimotor processing. In this paper, we present a method for mathematically modeling the changes in regional cerebral blood flow (rCBF) as a function of experimental parameters using step and linear functions. PET was used to measure rCBF in six subjects who tracked a target moving with constant amplitude across a computer screen at four different frequencies. Each subject tracked the target by flexing and extending the wrist. Two scans were performed at each frequency. The data for each subject were normalized by the mean blood flow in each scan and scaled to the mean blood flow at rest. Scaled rCBF was regressed onto movement frequency to identify voxels which had either a significant linear or step function response to the frequency of movement. A group analysis was also performed to identify significant functional changes common to all subjects. Significant rCBF increases in relation to movement frequency were found in the supplementary motor area, primary motor cortex, premotor cortex, thalamus, and cerebellum and localized using the Talairach atlas. Habituation of responses was not observed. PMID- 9343613 TI - I. PET studies of memory: novel and practiced free recall of complex narratives. AB - Positron Emission Tomography (PET) with the tracer H215O was used to measure regional cerebral blood flow in 13 healthy volunteers during two experimental memory tasks, one of which was well-practiced and the other of which was novel. The materials used for the memory tasks consisted of two complex narratives (Story A and Story B from the Wechsler Memory Scale). Natural language materials were chosen because they similate experimentally the natural learning situation and permit study of the neural mechanisms by which recall memory becomes more fluid, automatic, or "rote." One week before the PET study, subjects were trained to perfect recall of Story A, while they were exposed to Story B only 60 s prior to PET data acquisition. Despite the substantial differences in level of familiarity (and in free recall performance), patterns of activation were quite similar; activations presumed to reflect recall in both tasks included frontal, inferior temporal, thalamic, anterior cingulate, and cerebellar regions. Many regions were smaller during recall of the familiar story, however, presumably reflecting greater neural efficiency due to practice. In addition, the novel task activated an additional left frontal region that is presumed to reflect more active encoding. The similarity and multiplicity of the activations in the two tasks suggest that the brain uses a multinodal general network for memory tasks such as free recall, while the differences suggest that some nodes in the network may be used for specific components of memory such as encoding and retrieval. PMID- 9343614 TI - II. PET studies of memory: novel versus practiced free recall of word lists. AB - Positron emission tomography (PET) with the tracer H215O was used to measure regional cerebral blood flow in 13 healthy volunteers while they engaged in free recall of 15-item word lists from the Rey Auditory Verbal Learning task. The study was designed so that recall of well-practiced versus novel material could be compared. One week before the PET study, subjects were trained to perfect recall of List A, while they were exposed to list B only 60 s prior to PET data acquisition. As in the companion study of free recall of complex narratives, we observed that practice tended to decrease the size of activations in regions involved in the memory component of the task; we also observed that the novel recall task produced greater activation in left frontal regions, probably due to active encoding. A commonality of other regions observed in this pair of studies, as well as other studies of memory in the literature, suggests that the human brain may contain a distributed multinodal general memory system. Nodes on this network include the frontal, parietal, and temporal cortices, the thalamus, the anterior and posterior cingulate, the precuneus, and the cerebellum. There appears to be a commonality of components across tasks (e.g., retrieval, encoding) that is independent of content, as well as differentiation of some components that may be content-specific or tasks-specific. In addition, these results support a significant role for the cerebellum in cognitive functions such as memory. PMID- 9343615 TI - Blood flow in human anterior temporal cortex decreases with stimulus familiarity. AB - In this positron emission tomography investigation of human nonverbal visual memory, we determined the anatomical distribution of changes in regional cerebral blood flow (rCBF) reflecting familiarity of visual shapes. All stimuli consisted of relatively simple, abstract outlines generated by Fourier expansions. Subjects performed the same task twice, once using unfamiliar and once using familiar stimuli. The task was a modified version of delayed nonmatching-to-sample, consisting of one sample presentation prior to tracer injection and a sequence of test trials during which images were acquired. When this task was performed with familiar instead of unfamiliar stimuli, only decreases in rCBF were observed, which were localized to the left lateral anterior temporal neocortex, the left medial temporal pole, and the rostral anterior cingulate. The correlates of familiarity measured in this human brain mapping experiment may correspond with what has been measured in single neurons in monkeys. This correspondence holds both for the type and for the localization of changes. PMID- 9343616 TI - Organized systems of care. PMID- 9343617 TI - Usefulness of intrinsic infection risk indexes as predictors of in-hospital death. AB - OBJECTIVE: Comparison of two measures of intrinsic infection risk for predicting in-hospital mortality risks among subjects undergoing general surgery: the Study on the Efficacy of Nosocomial Infection Control (SENIC) index and the National Nosocomial Infection Surveillance (NNIS) index. DESIGN: Prospective cohort study on 1483 patients admitted to the service of general surgery of a tertiary hospital. The main outcome measure was in-hospital death. Relative risks, crude and multiple-risk factor adjusted for by logistic regression analysis, and their 95% CIs were estimated. RESULTS: During follow-up, 33 patients (2.2%) died. Both the SENIC and the NNIS indexes appeared related to in-hospital mortality risk in crude data. After several confounders (age, sex, severity of illness, American Society of Anesthesiologists score, serum creatinine, serum albumin, stay at the intensive care unit, length of operation, type of surgical wound, and preoperative stay) were controlled for, the SENIC index showed a borderline significant trend with mortality (p = 0.052), whereas the trend was significant for the NNIS index (p = 0.026). The NNIS index also showed a linear trend with both crude and adjusted for (SENIC index) risk of death. The SENIC index did not exhibit any linear trend with adjusted for (NNIS index) risk of surgical wound infection. To delineate whether the SENIC index added explanatory information to the NNIS index (or vice versa), we regressed the SENIC index on the NNIS index (and vice versa) and computed a set of residuals for both indexes. In logistic regression analyses, the residuals of NNIS index added meaningful information to the SENIC index, whereas the residuals of the SENIC index did not add any relevant information to the NNIS index. These results remained unchanged after controlling for several confounders. CONCLUSIONS: Both the SENIC and the NNIS indexes are good predictors of in-hospital mortality risk. The NNIS index had greater capability for discriminating and predicting risk of dealth. PMID- 9343618 TI - Epidemiology and control of vancomycin-resistant enterococci in an adult and children's hospital. AB - BACKGROUND: The incidence of vancomycin-resistant enterococci (VRE) has reached endemic proportions in many medical centers. To initiate an effective infection control program, an understanding of the epidemiologic attributes of the genus in medical facilities is imperative. METHODS: We studied 138 consecutive cases of VRE from April through December 1995. We created a database to analyze the risk factors for patients in both an adult hospital and a children's hospital and screened all specimens, submitted for routine microbiologic analysis, for VRE. RESULTS: One hundred twenty-three cases (89%) occurred in the adult acute care hospital, and 15 (11%) occurred in the children's hospital. Eighty patients (58%) were colonized with VRE, and 58 (42%) had an infection with VRE. Eighty-three percent of all the cases of VRE were nosocomially acquired. The majority of cases occurred in the medical service. Urine was the most important clinical specimen infected or colonized. Prior use of an antibiotic, other than vancomycin, was the most important risk factor for all nosocomial cases, followed by prior vancomycin use for surgical patients and residence in a unit with other patients infected with VRE for the medical service. Direct admission from another hospital was the most important risk factor for community-acquired cases. Special microbiologic screening of cultures yielded 48% of all VRE identified. Enterococcus faecium was the predominant resistant isolate recovered. CONCLUSIONS: The control of VRE in the hospital setting is difficult for several reasons. Almost half of all patients carrying VRE would not have been identified without special microbiologic screening efforts, as would patients, admitted from the community, who are already colonized with VRE. Controlling antibiotic use both in the hospital and the community is basic for controlling these organisms. Continuous education of all staff about VRE and other nosocomially significant organisms is the key to controlling the spread of these bacteria. PMID- 9343619 TI - Microbiologic evaluation of needleless and needle-access devices. AB - OBJECTIVE: This study was carried out to determine whether needleless intravenous access devices are more likely to allow microorganisms to enter the fluid pathway than intravenous needle-access devices. METHODS: A laboratory study was conducted with two needleless and one intravenous needle-access devices and Enterococcus faecium as a bacterial challenge. Inocula of E. faecium were prepared on the basis of the numerical estimates of 1000 to 10,000 colony-forming units (CFU)/cm2 of bacterial flora on dry regions of skin (arms, legs, and hands). The septum of each access device was inoculated with 10 to 20 microliters of a 10(4) to 10(5) CFU/ml challenge suspension, which was allowed to dry on the surface of the septum. In the first part of the experiment, the needleless or needle-access cannula of each device was used to puncture the corresponding septum without previously disinfecting the top of the septum. In the second part, the contaminated septum was punctured after disinfecting the septum with a 70% isopropyl alcohol wipe. After each puncture, trypticase soy broth was flushed through the fluid pathway of the intravenous access device, collected, and cultured by the membrane filtration technique. The septum of each injection-site cap and the needleless or needle-access cannula were sampled with sterile premoistened swabs. Swabs were cultured on blood agar plates. RESULTS: The rate of fluid pathway contamination was 100% (40/40) for one of the needleless intravenous access devices and 80% (20/25) for the other when septa were contaminated with E. faecium and not disinfected before puncture. The rate for the intravenous needle-access device was 72% (18/25). When the septa of the three different devices tested were disinfected with 70% isopropyl alcohol, E. faecium was isolated on only one septum from all devices tested in part two (1/74, 1.3%). CONCLUSIONS: These laboratory studies demonstrate that there is no statistically significant difference in the rate of fluid pathway contamination between needleless and intravenous needle-access devices. However, if the septa of either needleless or needle systems are not disinfected before puncture, a high rate of fluid pathway contamination may occur. PMID- 9343620 TI - Alcohol abuse: a risk factor for surgical wound infections? AB - BACKGROUND: The incidence of postoperative surgical site infections (SSIs) is difficult to estimate because of the current trend of early discharge after surgery. Both operation-related and host factors should be taken into consideration in the prevention of SSIs. We wanted to determine the actual incidence of SSIs and evaluate the risk factors in our clinic, using an extended follow-up period of 30 days after operations. METHODS: We performed a prospective follow-up survey of SSIs over a 3.5-month period including a 1-month follow-up after discharge with written instructions and a telephone survey. The SSIs were defined according to Centers for Disease Control and Prevention criteria. Forty three patient parameters were recorded, and risk factors for SSI were sought and tested by using multiple logistic regression analysis. RESULTS: The follow-up was completed in 772 of 807 patients. The SSI rates in these patients were 5.3% in clean, 7.1% in clean-contaminated, 6.2% in contaminated, and 28.1% in dirty operations. Seventy-one percent of infections were not diagnosed until after discharge from the hospital. According to multiple logistic regression analysis, alcohol abuse (p < 0.0001), wound contamination class (p < 0.05), and operation duration of over 2 hours (p < 0.05) were independently significant risk factors for SSI. CONCLUSIONS: A major portion of SSIs are found only after follow-up is extended during the postdischarge period. Alcohol abuse is a significant risk factor for SSI and should be taken into account when determining the susceptibility of an individual patient. PMID- 9343621 TI - Novel uses of the aromagram in infection control and epidemiology. AB - BACKGROUND: To facilitate the interpretation of data used in infection control and epidemiology, a novel data presentation format (the aromagram) has been developed and modified. METHODS: Aromagrams were developed with a personal computer-based graphics application. Aromagrams were based on antimicrobial susceptibility data from all specimen submitted to the University of California San Diego Medical Center's clinical microbiology laboratory between July 1992 and December 1994. RESULTS: The aromagrams created displayed both bacterial species specific and antimicrobial agent-specific susceptibilities. Additional modified aromagrams incorporated costs of antimicrobial agents and temporal trends in susceptibility of individual species to selected antibiotics. CONCLUSIONS: The aromagram is a unique format for data presentation that can be used to illustrate antimicrobial susceptibilities (specific to both organisms and antimicrobial agents), temporal trends in susceptibility data, and antimicrobial costs. Aromagrams may be used to display data useful to infection control and epidemiology professionals and to clinicians. PMID- 9343622 TI - Adjunctive use of monthly physician questionnaires for surveillance of surgical site infections after hospital discharge and in ambulatory surgical patients: report of a seven-year experience. AB - We report our experience with the use of monthly physician questionnaires, in conjunction with traditional in-house monitoring, for surveillance of surgical site infections (SSIs) in inpatients after hospital discharge and in ambulatory surgical patients (i.e., those not requiring perioperative hospitalization) over a 7-year period (July 1988 to June 1995) involving 156,977 surgical procedures. The mean annual response rate was 73% and did not change significantly from year to year (range, 71% to 75%), but the proportion of surgical procedures covered by returned surveys increased during the study period from 75% to 81% in inpatients and from 78% to 86% in ambulatory surgical patients (p < 0.0001 for both comparisons). Of 1051 SSIs identified, 231 (22%) were identified solely by the survey: 16% of SSIs in inpatients after discharge and 66% of SSIs in ambulatory surgical patients. Of 787 cases meeting the criteria for SSI on the basis of in house surveillance and listed on returned questionnaires, 366 (47%) were not marked as SSIs by the responding surgeons. We conclude that since its implementation in 1988, monthly physician surveys at our medical center continue to contribute significantly to identification of otherwise undetected SSIs. However, monthly questionnaires should only complement, not replace, traditional in-house surveillance. PMID- 9343623 TI - Secular trends in bloodstream infection caused by antimicrobial-resistant bacteria in New Jersey hospitals, 1991 to 1995. AB - INTRODUCTION: Antimicrobial resistance among bacteria is an increasing public health problem. In 1991, New Jersey was the first state to establish statewide, hospital-based surveillance for antimicrobial-resistant bacteria. METHODS: Each month, all 96 nonfederal New Jersey hospital laboratories complete a form listing the species identity and drug susceptibility results for selected antimicrobial resistant bacteria isolated from blood cultures from hospital inpatients. Penicillin-resistant Streptococcus pneumoniae and aminoglycoside-resistant gram negative rods were studied from 1991 to 1995. Vancomycin-resistant enterococci and imipenem-resistant gram-negative rods were studied from 1992 through 1995. RESULTS: From 1992 to 1995, the vancomycin-resistant enterococci bloodstream infection prevalence rate increased from 11 to 29 per 100,000 hospital admissions (p < 0.001); the rate was higher at larger hospitals, urban and inner-city hospitals, and teaching hospitals. From 1991 to 1995, the penicillin-resistant S. pneumoniae bloodstream infection rate increased from 1.1 to 9.9 per 100,000 admissions (p < 0.001). In contrast, bloodstream infection rates did not change significantly for imipenem-resistant (12.5 during 1992 and 14.1 during 1995, p = 0.4) or aminoglycoside-resistant (8.0 during 1991 and 6.8 during 1995, p = 0.4) gram-negative rods. CONCLUSIONS: We found that vancomycin-resistant enterococci and penicillin-resistant S. pneumoniae, but neither of two groups of antimicrobial-resistant gram-negative rods, are increasing rapidly in prevalence in New Jersey. Continued monitoring and interventions to slow these increases are needed. PMID- 9343624 TI - Use of scrubs and related apparel in health care facilities. AB - There is no scientific evidence that the use of scrubs or other related apparel contributes to either the cause or the prevention of infections associated with health care facilities. However, because this type of apparel is now used so commonly as a replacement for the more traditional type of uniform, its original function as an ensemble worn by surgical personnel no longer prevails. The variety of these applications actually raises a new series of issues and challenges that would be best resolved by a multidisciplinary health care group with representatives from administration, nursing, medicine, materials management, human resources, infection control, and other affected departments. The purpose of this report is to facilitate this process. PMID- 9343625 TI - Infections in solid-organ transplant recipients. PMID- 9343626 TI - Age-related hepatitis B seroconversion rates in health care workers. AB - BACKGROUND: Protective hepatitis titers are reported for more than 90% of healthy adults who received three intradeltoid injections of vaccine. Some factors that influence seroconversion rates include age, sex, and presence of chronic diseases. METHODS: Because of work-related factors that placed them at risk of acquiring hepatitis B, 112 employees, who ranged in age from 20 to 70 years with a mean age of 39.2 years, completed the hepatitis B vaccination series between 1986 and 1993. All participants received three vaccinations. RESULTS: Hepatitis B surface antibody did not develop in 16 of 112 recipients (14.2%, 95% CI, 7.6% to 20.8%). Race, sex, and duration to antibody titer did not affect rates of seroconversion. Age greater than 50 years was associated with significantly decreased seroconversion rates (64.7%, 95% CI, 42.0% to 87.4%) compared with seroconversion rates of those younger than 50 years of age (89.5%, 95% CI 83.3% to 95.7%, p = 0.02). CONCLUSIONS: Our results indicate that when a hepatitis B immunization program is implemented, seroconversion rates are lower than published rates for healthy adults and adolescents. We recommend that seroconversion data from immunization programs for employees at risk for hepatitis B be reviewed and that postimmunization testing be considered to ensure adequate protection for those employees at highest risk for nonconversion. PMID- 9343627 TI - Epidemiology of hepatitis B vaccine acceptance among urban paramedics and emergency medical technicians. AB - BACKGROUND: The epidemiologic pattern of hepatitis B vaccination acceptance has not been thoroughly examined in medical first responders. METHODS: A blood-borne pathogen questionnaire was administered to 255 paramedics and emergency medical technicians (EMTs) in a large, urban fire department. RESULTS: The overall prevalence of hepatitis B vaccination was 78%. The most frequently cited reason for not getting vaccinated was fear of contracting the hepatitis B virus from the vaccination (26%). Vaccination scheduling difficulties (23%) and lack of time to get vaccinated (20%) were also cited. Increased age, being an EMT, and not having obtained the rank of officer were independently and significantly associated with not having been vaccinated. CONCLUSIONS: Educational campaigns for medical first responders are needed to increase vaccination compliance, with a special emphasis on older workers and EMTs. Administrative barriers, such as vaccination scheduling difficulties, should also be addressed. PMID- 9343628 TI - Who washes hands after using the bathroom? AB - Handwashing is one of the most important control measures for preventing the spread of bacteria. Although young children are taught the procedure through different types of behavior modification, its effect has not been measured in older children. We have documentation that adults and health care workers have a compliance rate of only 50% with this basic control measure. This article reports on the compliance rate, duration, and handwashing techniques used by middle and high school students after using the bathroom. PMID- 9343629 TI - Effectiveness of computer-assisted instruction in increasing the rate of universal precautions--related behaviors. AB - BACKGROUND: With widespread noncompliance to universal precautions well established, an experimental study was designed to compare the rate of universal precautions--related behaviors between nurses who participate in computer assisted instruction. This study also explored the relationship between rates of universal precautions--related behaviors and subjects' demographic and experiential characteristics and history of occupational blood-borne exposure. METHODS: Data were collected by using a questionnaire to elicit information as to subjects' demographic and experiential characteristics and history of occupational blood-borne exposure. The Universal Precautions Assessment Tool was used to gather data on rates of universal precautions--related behaviors on two groups of registered nurses with 30 subjects per group. RESULTS: By using analysis of variance, the null hypothesis was rejected. The intervention used in this study did increase universal precautions--related behaviors. Multiple regression was used to analyze the research question and none of the variables were significant. Forty (67.8%) subjects reported receiving a needlestick or cut caused by a needle or sharp that was actually or potentially contaminated with blood or body fluids. Of these exposures, only one patient was known to be HIV antibody positive. CONCLUSION: Replication studies using computer-assisted instruction interventions are needed as are studies aimed at exploring other potentially effective interventions. PMID- 9343630 TI - Preventing transmission of blood-borne pathogens: a compelling argument for effective device-selection strategies. AB - Disease transmission from percutaneous injury occurs in 2% to 40% of health care workers (HCWs) after exposure to the hepatitis B virus (HBV), in 3% to 10% after exposure to the hepatitis C (HCV) virus, and in 0.2% to 0.5% after exposure to the HIV virus. According to a recently published case-control study from the Centers for Disease Control and Prevention, the following factors increase the risk of HIV seroconversion in HCWs after percutaneous exposure to HIV-infected blood: deep injury, visible blood on the device, procedures involving needle placement directly into a vein or artery, and terminal AIDS in the source patient. Postexposure use of zidovudine by HCWs appears to reduce the risk of HIV transmission by 79%. Institutions seeking to reduce the risk of HCW seroconversion should conduct analyses of specific tasks associated with these high-risk factors, and safety interventions should be installed when tasks and devices increase the risk of seroconversion. Although this type of outcome-based strategy may not significantly reduce the total number of needlestick injuries, reducing high-risk exposures minimizes disease transmission and maximizes the cost-effectiveness of the intervention. PMID- 9343631 TI - Evaluation of the acceptability of a needleless vascular-access system by nurses. AB - BACKGROUND: Needleless intravenous-access devices have been introduced in an effort to reduce needlestick injuries and possible transmission of blood-borne pathogens to health care workers. However, there are no data on the acceptance of these devices by nursing personnel. METHODS: A survey of nursing personnel was taken at Indiana University Medical Center after introduction of a needleless intravenous device to determine their opinion after use of the needleless device. RESULTS: The majority of the nurses (72 of 94, 70%) had a favorable overall opinion of the device. Among those with a favorable opinion, 76% (55/72) responded that reduced risk of needlestick injury was the most important reason. Among those who had a negative opinion about the needleless-device system, 32% (7/22) reported that contamination risk was their major concern. Those who were trained before device use were more likely to properly use and maintain the needleless intravenous-access system. Of 89 respondents, 75.3% (67/89) believed that the initial training was adequate; however, 43% (29/67) thought that additional training after using the device for some time would have been beneficial. CONCLUSIONS: Comprehensive education programs that include training before and after device use are necessary if new needleless intravenous-access systems are to be successfully introduced and accepted by nursing personnel. PMID- 9343632 TI - Could antibiotic-resistant pathogens be cross-resistant to hard-surface disinfectants? PMID- 9343633 TI - Sexual orientation differences in cerebral asymmetry and in the performance of sexually dimorphic cognitive and motor tasks. AB - With each of the tasks in the present studies we expected to find the reported sex difference between heterosexual women and heterosexual men and we predicted a sexual orientation effect with the performance of homosexual men being similar to that of heterosexual women and different from that of heterosexual men. Study 1 aimed to replicate earlier findings by recording the performance of a group of homosexual men on a visuospatial task, the Vincent Mechanical Diagrams Test (VMDT), a dot detection divided visual field measure of functional cerebral asymmetry, and on five subtests of the Wechsler Adult Intelligence Scale (WAIS). For each task the profile of scores obtained for the homosexual men was similar to that of heterosexual women in that they scored lower than heterosexual men on the VMDT, they showed less asymmetry, and they recorded a higher Verbal than Performance IQ on the WAIS. In Study 2, a male-biased targeted throwing task favored heterosexual men while, in contrast, on the female-biased Purdue Pegboard single peg condition heterosexual men were outperformed by heterosexual women and homosexual men. On neither of these two tasks did the performances of homosexual men and heterosexual women differ. One task, manual speed, yielded neither sex nor sexual orientation differences. Another, the Purdue Pegboard assemblies condition, revealed a sex difference but no sexual orientation difference. Failure to obtain a sexual orientation difference in the presence of a sex difference suggests that the sexual orientation effect may be restricted to a subset of sexually dimorphic tasks. PMID- 9343634 TI - Understanding sexual coercion among young adolescents: communicative clarity, pressure, and acceptance. AB - Young people's understanding of sexual coercion was studied. Boys and girls (N = 191) were asked to rate scenarios depicting sexual situations according to their perceptions of communicative clarity, the extent of pressure being applied to one partner, and the acceptability of the behaviors. Judgments of communicative clarity were given more readily when there was consent rather than dissent to sex. Clear communication was readily inferred even when there were no cues that this was the case. Boundaries of behaviors that were defined as constituting "pressure" were influenced by the outcome, that is whether sex did or did not occur, as well as the behavior itself. Ratings of acceptability closely followed those of pressure, although the relationships between perceptions of pressure and acceptability were stronger for girls than for boys. In general, there were few gender differences in perceptions of pressure and communicative clarity. Of concern was the finding that, for some respondents, pressure and acceptability were unrelated to the use of either physical or emotional force. PMID- 9343635 TI - Testosterone treatment in men with erectile disorder and low levels of total testosterone in serum. AB - Since decreased serum levels of testosterone (T) do not necessarily predict good outcome of testosterone treatment for erectile disorder, the purpose, of this study was to determine which men with erectile disorder and decreased serum levels might benefit from treatment. From a sample of 31 men (mean age = 39 years), 15 (48%) with erectile disorder and decreased serum levels of T responded well after 8 weeks of testosterone treatment (100 mg of testosterone propionate in the sustained-release form given im once a week). Good treatment outcome was associated with several variables, but only high levels of luteinizing hormone (LH) and low values of the T/LH (testosterone/LH) ratio consistently emerged as significant correlates and/or predictors of effective treatment. Levels of LH above 7.5 IU/L or the values of the T/LH ratio equal to or below 0.87 nmol/IU in patients with erectile disorder and decreased serum levels of T suggest that testosterone treatment may be effective. PMID- 9343636 TI - Hypersexual desire in males: an operational definition and clinical implications for males with paraphilias and paraphilia-related disorders. AB - The longitudinal history and temporal stability of total sexual outlet (TSO) in a group of outpatient males with paraphilias (PA) and paraphilia-related disorders (PRD) was assessed. Based on extant normative data from contemporary population based surveys of sexual behavior, it was hypothesized that a persistent TSO of 7 or more orgasms/week for a minimum duration of 6 months be considered as the lower boundary for hypersexual desire in males. In almost all statistical analyses, the PA (n = 65) and PRD (n = 35) groups were not statistically different. The mean current TSO (PA, 7.4 +/- 5.7; PRD, 8.0 +/- 4.2) as well as the current average time consumed in all unconventional sexual behaviors (1-2 hr/day) were not statistically different. Unconventional sexual behaviors (i.e., related to PAs or PRDs) leading to orgasm constituted 77% of current TSO. In the combined group (n = 100), 72% (n = 72) reported a hypersexual TSO of 7 or greater. Age of onset of hypersexual TSO in the PAs (19.2 +/- 6.8 years; range 10 43) and the PRDs (21.0 +/- 8.6; range 10-46) and the duration of hypersexual TSO (PA, 11.1 +/- 11.2 years; PRD, 10.5 +/- 9.1) were not significantly different. Fifty-seven males (57%) reported a TSO of 7 or more for a minimum duration of 5 years. Clinical implications of reconceptualizing PAs and PRD as sexual desire disorders are discussed. PMID- 9343637 TI - Treatment outcome of brief couple therapy in psychogenic male erectile disorder. AB - Treatment outcome was studied in 37 couples who entered brief combined sex and relationship therapy for male erectile disorder in a specialized clinic. Treatment was completed by nearly two thirds of the couples. Significant improvements in target symptoms, questionnaire scores, including the Golombok Rust Inventory of Sexual Satisfaction, and frequency of attempts at sexual activity were recorded. Results suggest that behavioral-systems couple therapy and modified modern sex therapy offer a brief, flexible, and reproducible treatment option for men with psychogenic factors associated with erectile disorder. PMID- 9343638 TI - Coming out by south Asian gay men in the United Kingdom. AB - The process of coming out among Western gay men and women is well described. The present study is the first to explore the experiences of coming out among gay men of South Asian origin in the U.K. South Asian is defined here as originating from the Indian subcontinent. Members of a homophile organization were given a questionnaire designed to assess the experiences of coming out to family and friends and the degree of compartmentalization in their lives. Information obtained from 52 questionnaires was supplemented by detailed interviews with 9 respondents. Families and religion played important roles in the process of coming out. Sisters were most likely to be told first. Some degree of dissonance between cultural and sexual identity was noted. In addition, each step taken in revealing one's sexual orientation to friends, family, and colleagues was dictated by the strength of the relationships and the desire for intimacy. Further areas of research are highlighted. PMID- 9343639 TI - Research suggests that the sex ratio (proportion male) of siblings of some samples of male homosexuals may be high. PMID- 9343640 TI - Discussion concerning maternal testosterone levels in pregnancy. PMID- 9343642 TI - Arthroscopic meniscal repair using fibrin glue. Part II: Clinical applications. AB - Since 1984 we have arthroscopically repaired meniscal tears using a purified fibrin glue. This article describes clinical application of a fibrin-based glue to arthroscopic meniscal repair and a long-term follow-up study of 61 repaired menisci in 40 patients (average follow-up, 8 years; range, 5.1 to 11.4 years). Of these patients, 6 complained of recurrent meniscal symptoms and received partial meniscectomy. According to stepwise logistic regression analysis, the factors most strongly correlated with recurrent symptoms were insufficiency of the associated anterior cruciate ligament, repairs of fresh tears, and repairs requiring supplementary meniscal sutures (P < .05). PMID- 9343641 TI - Arthroscopic meniscal repair using fibrin glue. Part I: Experimental study. AB - An experimental study of rabbit menisci was carried out to evaluate the healing promoting properties of fibrin glue and fibrin glue-containing marrow cells. A full-thickness defect, 1.5 mm in diameter, was made within the avascular portion of the meniscus and left empty in 20 menisci (C group), filled with fibrin glue in 20 menisci (F group), and filled with fibrin glue-containing marrow cells in 20 menisci (M group). Measurements of the remaining defects and histological examinations were performed 1, 3, 6, and 12 weeks after each procedure. Overall, the remaining defects in the F group and, particularly, in the M group were significantly smaller than those in the C group at the various time points. Furthermore, the results of histological study showed earlier mature healing of the defects in the M group than of those in the F group. Our results suggest that fibrin glue, especially in a preparation containing marrow cells, may enhance meniscal healing. PMID- 9343643 TI - Chronic pain following ankle sprains in athletes: the role of arthroscopic surgery. AB - We reviewed 100 patients treated arthroscopically for symptoms of chronic ankle pain associated with sprains of the ankle. All had pain that had failed to respond to conservative treatment for at least 6 months. The pathology in 95 of the 100 ankles studied could be categorized into one of three groups: the instabilities (lateral and syndesmotic), the impingements (anterior and anterolateral), and articular lesions (chondral and osteochondral). Five patients had nonspecific osteoarthritis and/or synovitis on arthroscopy. Patients were followed-up for improvements in six categories: pain, swelling, stiffness, limping, activity, and instability. The primary outcomes of pain and activity were analyzed statistically. Patient satisfaction and return to sports were evaluated. Significant improvements were obtained for patients treated for syndesmotic instability, and anterior and anterolateral impingement. Chondral fractures in the presence of a stable ankle had good results in 75% of cases, compared with those in unstable ankles with only 33% good results. Osteochondritis dissecans was treated successfully by excision of the lesion and abrasion of the base. Patients with chronic lateral instability were treated by open repair, so only the diagnostic arthroscopic findings are reported. We concluded that arthroscopy offered little to the management of lateral instability unless there was considerable doubt regarding the diagnosis. There were minimal improvements for the patients with nonspecific diagnoses such as posttraumatic synovitis. Ankle arthroscopy may be a very useful diagnostic and therapeutic tool in patients who have not responded to conservative therapy. PMID- 9343644 TI - The ligamentum teres of the hip: an arthroscopic classification of its pathology. AB - Pathology of the ligamentum teres is rarely diagnosed. We describe the classification of the lesions seen at hip arthroscopy based on a group of 20 patients. Three groups are noted: complete rupture, partial rupture, and the degenerate ligamentum. The complete ligamentum teres rupture group had a history of either major trauma or surgery and had a high incidence of other hip pathology such as labral tears and articular damage. The partial ligamentum rupture group presented with a long history of ill-defined hip pain. Minor associated hip abnormalities were seen at arthroscopy. Degenerate ligamentum teres rupture presented with symptoms of the underlying osteoarthritis. Debridement or washout was performed but, at 2 years, patients had severe persistent symptoms or had had a joint replacement. PMID- 9343645 TI - Transplantation of fresh-frozen menisci: an experimental study in dogs. AB - Transplantation of 25 fresh-frozen medial menisci was studied in 15 adult dogs. Before implantation, the allografts were deep-frozen and stored at -70 degrees C for 6 weeks to 18 months. The animals were killed 2 to 8 months postoperatively, and their knees and transplants were examined macroscopically and histologically. Complete healing of the allografts was found in 19 knees, incomplete in 3, and healing by massive fibrovascular tissue in 3 knees. Some shrinkage of the transplants taken 2 to 4 months after the surgery was observed; however, the 6- and 8-month specimens appeared grossly normal. Histologically, all transplants displayed a characteristic decrease in the number of cells, but this was significantly less pronounced in the 6- and 8-month specimens. Some degenerative alterations were found in all transplanted knees, but was obviously less pronounced in areas covered by the allografts and in the 6- and 8-month specimens. It is concluded that the transplantation of the fresh-frozen menisci could be successful; although the transplants are subjected to a remodeling process, they appear to function normally and protect the articular cartilage. The technique of conservation by freezing at -70 degrees C offers the advantage of an effective meniscal banking. PMID- 9343646 TI - The efficacy of intra-articular morphine for postoperative knee arthroscopy analgesia. AB - This article describes two prospective, randomized, double-blind clinical trials designed to investigate this. Trial 1 compared a conventional local anaesthetic agent (100 mg bupivacaine) injected intra-articularly (i.a.) with a control (normal saline) and 1 mg of i.a. morphine. No significant difference was noted in the first 4 hours between the groups with respect to visual analogue pain (VAS) scores. However, at 6 and 24 hours, the group of patients who received 1 mg i.a. morphine recorded lower pain scores and required less supplementary analgesia. Trial 2 assessed the dose response relationship for i.a. morphine comparing 5 mg intravenous (i.v.) morphine (control) with 1 mg and 5 mg i.a. morphine. At early time points (1, 2, and 4 hours) similar VAS pain scores were recorded for both 5 mg i.v. morphine and 5 mg i.a. morphine, both significantly lower than the group receiving 1 mg i.a. morphine. At 6 and 24 hours, 5 mg of i.a. morphine produced significantly lower pain scores, less analgesic requirement, and less sleep disturbance on the first postoperative night than the other groups. It can be concluded from these two studies that 5 mg i.a. was the most effective analgesic following knee arthroscopy. PMID- 9343647 TI - Popliteomeniscal fasciculi and the unstable lateral meniscus: clinical correlation and magnetic resonance diagnosis. AB - We hypothesize that disruption of the fascicular attachments between the popliteus and lateral meniscus can result in gross instability of the meniscus producing locking of the knee. This study brings attention to the importance of the clinical examination, and the need for clinical correlation to magnetic resonance (MR) studies. We report on three patients referred with the history of mechanical locking episodes of their knee. Initial MR examinations were all read as normal before referral to our institution. On close review of these MR examinations, popliteomeniscal fascicular disruption could be seen in each case. Each of these patients had arthroscopic-repair of these meniscal detachments. At 1-year follow-up, all patients had resolution of mechanical symptoms. Each patient had confirmation of their repair with repeat arthroscopy or MR and arthrographic examinations. An anatomic specimen was used to identify the popliteus muscle and tendon, the lateral meniscus, the antero-inferior popliteomeniscal fascicle, and the postero-superior popliteomeniscal fascicle attachments. MR images of the same anatomic specimen show both superior and inferior fasciculus attachments to the capsule. MR examples of the intact and disrupted antero-inferior and postero-superior popliteomeniscal fasciculi have been correlated to anatomic specimens to help familiarize the orthopaedic surgeon with these important stabilizing structures. PMID- 9343648 TI - Difficulties with the "N + 7 rule" in endoscopic anterior cruciate ligament reconstruction. AB - In an effort to facilitate the use of interference screw fixation in the tibia, a method to orient the tibial tunnel termed the "N + 7 Rule" was prospectively applied to 60 consecutive endoscopic anterior cruciate ligament reconstructions performed by a single surgeon. Four cases were excluded because a breach in the described protocol for application of the N + 7 rule was identified at the time of the operation. Of the remaining 56 cases, 28 (50%) were fixed with an interference screw on the tibia, whereas 28 (50%) required fixation with sutures tied over a post. The 28 cases not amenable to interference screw fixation included 17 cases in which the tibial tunel was too long and 11 cases in which the tunnel was too short. It appears that the applicability of the N + 7 rule to an individual surgeon's practice may be altered by small variations in operative technique. PMID- 9343649 TI - Morphology of the axillary nerve in an anteroinferior shoulder arthroscopy portal. AB - An anteroinferior portal can be safely used in arthroscopic shoulder surgery but requires an in-depth knowledge of axillary nerve anatomy. The purpose of this report is to present the qualitative and spatial anatomy of the axillary nerve and to describe patterns of arborization that may affect safe anteroinferior arthroscopic portal placement. Measurements were taken in 42 embalmed cadaveric shoulders (20 male, 22 female). The distance from the acromioclavicular (AC) joint to the axillary nerve averaged 7.90 cm (range, 7.2 to 9.1 cm) in males and 6.37 cm (range, 5.2 to 8.1 cm) in females. We describe the axillary nerve index (distance of nerve from the AC joint/length of deltoid from AC joint) which can be used to predict the location of the axillary nerve along the anterior clavicular line (ACL). The axillary nerve index averaged 0.48 (range, 0.42 to 0.57) in males and 0.41 (range, 0.31 to 0.57) in females. Four types of morphology were noted in the axillary nerve: (1) main trunk with superior and inferior branches, (2) main trunk with superior branches, (3) main trunk with inferior branches, and (4) main trunk only. Our work supports the traditional operable safe zone for the axillary nerve. PMID- 9343650 TI - Arthroscopic synovectomy in the management of painful localized post-traumatic synovitis of the knee joint. AB - Seventy-four patients were operated on by arthroscopic synovectomy after being diagnosed with symptomatic localized post-traumatic synovitis in a 5-year period. Sixty-six patients (69 knees) could be evaluated at the end of the follow-up. The indication for surgery was severe mechanical type pain (score 3) after an evident injury nonresponding to 3 months of conservative treatment. Any other source of pain was preoperatively excluded by means of radiographs and magnetic resonance imaging, and intraoperatively by means of arthroscopic examination. The results were assessed according to a pain scale (score 0-3). Preoperatively all patients had severe pain (score 3). After a mean follow-up of 2.8 years (range, 1 to 6 years) there were 43 patients without pain (score 0), 15 with mild pain (score 1), 8 with moderate pain (score 2), and 3 with severe pain (score 3). In summary, this retrospective study appears to show that arthroscopic synovectomy should be taken into account when facing patients with painful localized post-traumatic synovitis of the knee joint. This is a diagnosis of exclusion as other internal derangements have been eliminated by magnetic resonance imaging and arthroscopic examination. PMID- 9343651 TI - Arthroscopic transglenoid multiple suture repair: 2 to 8 year results in 150 shoulders. AB - One hundred fifty-six arthroscopic transglenoid multiple suture repairs were performed for chronic anterior shoulder instability. In 150 shoulders (96% follow up), the outcome with respect to recurrence of instability and the Bankart Score was determined a minimum of 2 years and a mean of 4.1 years after surgery (range, 2 to 8.2 years). During the follow-up interval, 11 shoulders (7.3%) redislocated. Fourteen other shoulders (9.3%) had at least one episode that we interpreted as recurrent subluxation. Shoulders with a Bankart lesion and younger patients had a higher probability of recurrent instability (P < .05). We concluded that this method is most effective in shoulders without a Bankart lesion and in patients older than 25 years of age (regardless of pathology). PMID- 9343652 TI - The fate of intra-articular debris following arthroscopic anterior cruciate ligament reconstruction. AB - The debris generated during arthroscopic anterior cruciate ligament (ACL) reconstruction may be seen on postoperative radiographs. The purpose of this study was to evaluate the incidence, effects, and natural history of intra articular debris following ACL reconstruction. This retrospective review included 99 ACL reconstructed knees in 96 patients. Radiographically visible debris was present in 63% of knees (bone in 59% and metal in 4%), and 37% of knees had no visible debris. Bone was most commonly seen in the posterior compartment (95%), and metal within the intercondylar notch. There were no differences in the incidence of debris between reaming techniques, single and dual incision techniques, or between graft types. Metal debris was always associated with retrograde reaming. Complete disappearance of bone debris was noted in 71% (3 to 6 months). Of the 25% of knees that showed persistent bone debris, in 79% it had decreased in size. There was no change in the appearance of metal debris (4%). No patient experienced mechanical symptoms directly related to debris. No secondary surgeries for debridement of debris or loose body removal were required. Bone debris produced during arthroscopic ACL reconstruction appears clinically benign, and is likely to disappear by 6 months. Long-term effects are unknown. Metal debris is persistent, but not problematic over the short-term. PMID- 9343653 TI - The outcome of operatively treated anterior cruciate ligament disruptions in the skeletally immature child. AB - The purpose of this study was to evaluate the outcome of transphyseal ligament reconstruction in skeletally immature children with midsubstance anterior cruciate ligament (ACL) disruption. Five consecutive patients (mean age, 12.9 years; range, 8 to 14 years) with radiographically documented "wide" open growth plates and a minimum of 5 cm of expected remaining growth, underwent intra articular reconstruction of the ACL. Operative treatment included three ACL reconstructions using hamstring tendons and two with quadriceps patellar tendon. All involved a centrally placed 6-mm or smaller tibial drill hole through an open physis and graft placement in an over-the-top position on the femur. At an average follow-up of 7.4 years (range, 4.5 to 9.9 years), no patient had a positive anterior drawer, Lachman, or pivot shift test. On KT-1000 arthrometer testing, all patients had 3 mm or less of increased anterior-posterior displacement (mean +/- SD = 1.0 +/- 1.6 mm). Magnetic resonance imaging showed that four tibial physes had fused in a symmetric fashion and one was still open. Orthoroentgenograms showed that no patient had a significant leg length discrepancy (-0.8 mm +/- 3.4 mm). The mean increase in height postoperatively was 17.7 cm (range, 7.6 to 31.0 cm). Overall, using the International Knee Documentation Committee (IKDC) evaluation form, there were four patients with grade A and one with grade C. The one patient with a poor IKDC grade had sustained a subsequent patellar dislocation with osteochondral fracture. In conclusion, ACL reconstruction using small drill holes placed through open tibial physes does not seem to adversely affect outcome or future growth. PMID- 9343654 TI - Laser-assisted shoulder surgery. AB - The application of laser/thermal energy in arthroscopic shoulder surgery remains controversial. Laser proponents tout the benefits of coagulation and vaporization of tissue, whereas opponents cite costs, complications, and the fact that the laser has not yet shown results superior to presently available mechanical techniques. A lack of basic science studies and the aversion of many physicians to the marketing aspects of laser technology have undermined the widespread orthopaedic acceptance of laser techniques. Newer applications, such as capsular shrinkage are just now being evaluated as to effect and efficacy. Orthopaedists should be assured that, at present, they are not compromising patient care by not using laser techniques. PMID- 9343655 TI - Combined tear of the posterior cruciate and medial collateral ligaments resulting in a locked knee. AB - A case is presented of a combined tear of the posterior cruciate and medial collateral ligaments with medial collateral ligament fibers herniated through the medial capsule and occupying the medial compartment, which resulting in a locked knee. The author is not aware of any reports in the literature of this and presents this to raise awareness of another cause of knee locking. PMID- 9343656 TI - Integration of hamstring tendon graft with bone in reconstruction of the anterior cruciate ligament. AB - Anterior cruciate ligament (ACL) reconstruction using four-strand hamstring graft with round-headed, cannulated, interference (RCI) screw fixation requires osteointegration of the tendon graft. This report describes the histology at the bone-tendon junction of two specimens retrieved from patients undergoing revision surgery after traumatic mid-substance ACL graft rupture at 6 and 10 weeks after initial reconstruction. Revision was performed at 12 and 15 weeks. Integration of the graft was evident by observation of collagen fiber continuity between bone and tendon. This histology plus the low incidence of early graft failure suggest that free tendon autograft attached to bone by RCI screw allows adequate osteointegration between 6 and 15 weeks after surgery. PMID- 9343657 TI - Pseudoaneurysm as a complication of knee arthroscopy. AB - This is a report of a pseudoaneurysm of the inferior medial geniculate artery following knee arthroscopy. This case was treated successfully with embolization. PMID- 9343658 TI - Compartment syndrome of the leg after arthroscopic examination of a tibial plateau fracture. Case report and review of the literature. AB - Increasing attention has been given to the role of arthroscopy in the treatment of tibial plateau fractures. Complications of arthroscopy are infrequent but are potentially severe. We present a case of acute compartment syndrome related to arthroscopic treatment of a tibial plateau fracture in an adolescent. PMID- 9343659 TI - Acromion-splitting approach through an os acromiale for repair of a massive rotator cuff tear. AB - Os acromiale, failure of fusion of the secondary centers of ossification of the acromion process, has been noted as a contributing factor in shoulder impingement syndrome and rotator cuff tears. Treatments for symptomatic os acromiale with or without rotator cuff tears have been reported in the literature and range from excision of small fragments to fusion of larger, fragments with internal fixation and bone grafting. Generally, rotator cuff repairs have been performed when possible. We report an acromion splitting approach through an existing os acromiale to gain exposure for the repair of a massive rotator cuff tear. Subsequent to this repair, the acromion was repaired with internal fixation. Good functional use of the patient's upper extremity was obtained and the patient expressed satisfaction with the surgical outcome. The acromion splitting approach is a viable approach in patients with an os acromiale and a coexistent rotator cuff tear. PMID- 9343660 TI - Arthroscopic anterior cruciate ligament reconstruction using autogenous hamstring tendon graft without detachment of the tibial insertion. AB - This article describes a modified arthroscopic technique of anterior cruciate ligament reconstruction using quadrupled hamstring tendon graft. The autogenous semitendinosus and gracilis grafts are harvested without detachment of the tibial insertion. To obtain longer grafts, the accessory tibial insertions of the hamstring tendons are dissected. The EndoButton (Acufex Microsurgical, Andover, MA) is used for femoral fixation and two spiked staples are used for tibial fixation in a belt buckle fashion. Then the residual anterior laxity is restored by additional absorbable interference screw fixations. In this technique, more viable graft is obtained and more firm distal fixation is achieved by preservation of the tibial insertion of hamstring tendons. PMID- 9343661 TI - Arthroscopic posterior cruciate ligament reconstruction using four-strand hamstring tendon graft and interference screws. AB - We describe an arthroscopic technique for reconstruction of the posterior cruciate ligament (PCL) using a four-strand hamstring tendon graft. The femoral tunnel is drilled via the anterolateral portal and the tibial tunnel through the skin incision from the graft harvest. The graft is pulled through the tunnels with pullout sutures and fastened with interference screws. The results to date are good and the procedure can often be performed as day surgery. PMID- 9343662 TI - Arthroscopic technique: two-piece excision of discoid meniscus. AB - This article describes a new arthroscopic technique of excision of complete discoid meniscus with a tear in which the discoid is divided into anterior and posterior pieces for removal. It is easy to perform because an excellent visualization of the posterior segment is obtained after removal of the anterior piece. The extent of intrasubstance pathology can be evaluated with a transverse cut to the mid portion of the discoid, and therefore, it is easy to determine the width of the rim to be retained. Because the meniscus is divided into two pieces, a large portal is not required for removal. PMID- 9343663 TI - Modification of a PCR-based site-directed mutagenesis method. PMID- 9343664 TI - Typing of human papillomaviruses by reductional RFLP analysis of biotin-labeled PCR fragments. PMID- 9343665 TI - Long-term storage of unamplified complete PCR mixtures. PMID- 9343666 TI - Preparation of DNA from numerous individual microscopic organisms for PCR-based assays of environmental samples. PMID- 9343667 TI - Site-directed mutagenesis: a two-step method using PCR and DpnI. PMID- 9343668 TI - Improved alkaline lysis method for rapid isolation of plasmid DNA. PMID- 9343669 TI - High-yield method for isolation of lambda DNA. PMID- 9343670 TI - Efficient recovery of plasmid DNA from Erwinia herbicola with high nuclease activity. PMID- 9343671 TI - Rapid preparation and identification of insert-containing recombinant plasmid DNA. PMID- 9343672 TI - Microgranular cellulose improves dsRNA recovery from plant nucleic acid extracts. PMID- 9343673 TI - Design and use of easily made RNA size markers. PMID- 9343674 TI - RT-PCR detection of RNA viruses in stool specimens. PMID- 9343675 TI - Vector for positive selection of in-frame genetic sequences. PMID- 9343677 TI - Simplified ELISA for detecting antibodies to recombinant fusion proteins. PMID- 9343678 TI - Generation of moderate amounts of polyclonal antibodies in mice. PMID- 9343676 TI - Baculoviral transfer vectors for expression of FLAG fusion proteins in insect cells. PMID- 9343679 TI - Hybridization and detection of biotinylated oligonucleotide probes in agarose gels. PMID- 9343680 TI - Improved electrophoretic separation of polymorphic short tandem repeats in agarose gels using bis-benzimide. PMID- 9343681 TI - High-temperature sodium dodecyl sulfate polyacrylamide gel electrophoresis. PMID- 9343682 TI - Safranin O counter-staining enhances the counting of beta-galactosidase expressing cells. PMID- 9343683 TI - Improved procedure for electroporation of peptides into adherent cells in situ. PMID- 9343684 TI - Simplified agar plate method for quantifying viable bacteria. PMID- 9343685 TI - Low-voltage electric-discharge biolistic device. PMID- 9343686 TI - Modification of an in situ renaturation method for analysis of protein kinase activity with multiple substrates. PMID- 9343687 TI - Efficient 5'-end labeling of oligonucleotides containing self-complementary sequences. PMID- 9343688 TI - Detection of major histocompatibility complex class I antigens on the surface of a single murine blastocyst by immuno-PCR. PMID- 9343689 TI - Method to improve reliability of random-amplified polymorphic DNA markers. PMID- 9343690 TI - Variable-size injectable dialysis chambers. PMID- 9343691 TI - Ethidium bromide enhances transformation of E. coli with homopurine/pyrimidine rich DNA. PMID- 9343692 TI - Comments on an article by A.M. Malek et al. PMID- 9343693 TI - Expression of green fluorescent protein in Aureobasidium pullulans and quantification of the fungus on leaf surfaces. AB - A red-shifted, mutated form of the jelly-fish green fluorescent protein (GFP) under control of a TEF promoter was expressed at high levels in the filamentous fungus Aureobasidium pullulans. In the three transformants studied, all morphotypes of the fungus, including pigmented chlamydospores, expressed GFP and fluoresced brightly. Confocal microscopy showed that the intra-cellular distribution of GFP was nonuniform. When applied to leaf surfaces, the transformants were readily visible and amenable to quantification by image analysis. Thus, GFP expression, together with quantitative image analysis, may provide a powerful method for ecological studies of plant-microbe relationships in nature. PMID- 9343694 TI - n beta geo, a combined selection and reporter gene for retroviral and transgenic studies. AB - Nuclear-targeted beta-galactosidase (beta-gal) is increasingly used as a genetic cell marker in vitro and in vivo. Nuclear sequestration concentrates beta-gal and permits sensitive identification of expressing cells and/or tissues without obscuring the cytoplasmic detail necessary for analysis of cell phenotype. Here, we report the construction and testing of a nuclear-targeted version of the beta geo fusion protein that combines nuclear localization with the ability to select expressing cells with the drug G418. This new marker gene functions efficiently in retroviral vectors and will be useful in identification and isolation of cells transfected in vitro and cells expressing transgenic or gene-targeted constructs in vivo. PMID- 9343695 TI - Flow cytometric quantification of surface-displayed recombinant receptors on staphylococci. AB - Surface display of recombinant proteins on bacteria and phages has become an important tool in bioscience. To evaluate the various host systems, a great need exists for quantitative methods to determine the densities of displayed proteins and peptides on the bacteria and phage surfaces. Here we describe how a method previously applied for quantification of surface proteins on mammalian cells has been adapted for quantification of chimeric receptors surface-displayed on bacteria; in this study, the bacteria being recombinant staphylococci. The presented method takes advantage of fluorescence-activated cell sorting (FACS) technology and a new type of nonfluorescent plastic beads, similar in size (2 microns in diameter) to bacterial cells, and thus suitable for generation of calibration curves from which the number of chimeric receptors can be obtained. The method was used to estimate the number of antigenic sites on two types of recombinant staphylococci, both carrying heterologous chimeric receptors, and it was found that the recombinant Staphylococcus carnosus cells carried approximately 10(4) surface-displayed antigenic sites, while recombinant Staphylococcus xylosus exposed approximately 3 x 10(3) sites per cell. The use of the deviced method for different applications is discussed. PMID- 9343696 TI - PCR mutagenesis-based method for generation of positive controls for SSCP analysis. AB - We have developed a primer-mediated PCR mutagenesis-based method for the generation of positive controls to test the sensitivity of single-strand conformation polymorphism (SSCP) or any other PCR-based mutation screening method. This technique is based on the incorporation of a third longer primer, containing a mismatched base, into the PCR along with the two wild-type primers normally used to amplify DNA fragments for SSCP analysis. The longer mismatch primer (LMP) shares the sequence of one of the wild-type primers and also contains 5 to 10 additional bases, which include the mismatched base. The resulting PCR product is identical in length and sequence to the wild-type template with the exception that the LMP base mismatch is incorporated into nearly 100% of the product. We have observed an altered SSCP mobility pattern in all cases where positive controls have been generated using this technique. We believe that the use of such in vitro-generated controls can contribute to the interpretation of band patterns and to the optimization of experimental conditions for SSCP to facilitate maximum detection of sequence variants. PMID- 9343697 TI - Competitive oligonucleotide single-base extension combined with mass spectrometric detection for mutation screening. AB - A rapid, robust and widely applicable mutation detection scheme not requiring radioactivity or gel-based detection is introduced. It is a single-tube, competitive oligonucleotide single-base extension (COSBE) reaction using a pair of primers with the 3'-terminal base complementary to either the normal or mutant allele. Upon hybridization and addition of a polymerase and the nucleotide triphosphate corresponding to the next base after the primer, only those primers properly annealed (i.e., no 3'-terminal mismatch) are extended; products are resolved by molecular weight shifts as determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (single-scan spectrum acquisition < < 1 s). For the cystic fibrosis delta F508 polymorphism, 28-mer "normal" (N) and 30-mer "mutant" (M) primers generate 29-mer (N + I) or 31-mer (M + 1) products for homozygotes and both for heterozygotes. Since primer and product molecular weights are relatively low (< 10 kDa) and the mass difference between these are at least that of a single approximately 300-Da nucleotide unit, a low-resolution mass spectrometer is suitable for such measurements. PMID- 9343698 TI - Specific inhibition of PCR by non-extendable oligonucleotides using a 5' to 3' exonuclease-deficient DNA polymerase. AB - The Stoffel fragment of Taq DNA polymerase lacks the 5' to 3' exonuclease activity that hydrolyzes potentially blocking DNA strands during primer extension. We there-fore asked whether by using this fragment in the PCR, non extendable, base-paired oligonucleotides could inhibit amplification in a sequence-dependent manner. Model targets were chosen from the partially conserved ribosomal 16S rDNA of three bacterial species: E. coli, Bacillus subtilis and Neisseria gonorrhoea. A single pair of primers was capable of amplifying a homologous 240-bp region from all three. Two non-extendable "blocking" oligonucleotides were synthesized with sequences complementary to the inter primer regions of E. coli and B. subtilis, respectively. Both blockers were shown specifically to prevent amplification of their complementary targets, but not of the reciprocal control targets or of the reciprocal control targets or of the non complementary N. gonorrhea. Specificity was further confirmed by an internal positive control. Similar inhibition was seen with mixtures of targets in a single reaction. With intact Taq DNA polymerase, no blocking was observed. Primers and blockers targeting specific regions of N. gonorrhoea rDNA were used to confirm the requirement that blockers be directed to the inter-primer region. Sequence-dependent amplification inhibition, such as that demonstrated here, would be applicable to PCR-related strategies using primers capable of using multiple targets, where such selective inhibition could be useful. PMID- 9343699 TI - Calibration and storage of DNA competitors used for contamination-protected competitive PCR. AB - DNA fragments used as standards in competitive PCR were precisely calibrated using HPLC and commercially available DNA molecular mass markers. The accuracy of calibration was reflected by data that differed by only 2% from the mean when two independently purified and calibrated competitor preparations were compared. Highly dilute competitor solutions were stable at -20 degrees C for up to 1 year in the presence of carrier HindIII-digested lambda DNA, but progressive loss of competitor DNA with increasing storage time was observed when carrier DNA was omitted from the solution. Applying 0.2 U uracil-DNA glycosylase (UDG) per assay of remaining temperature-stable activity did not effect the ratios of synthesized products. This study describes quality management in PCR quantitation that is useful for the measurement of multidrug resistance-associated protein (MRP) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene transcripts. PMID- 9343700 TI - Copper treatment increases recombinant baculovirus production and polyhedrin and p10 expression. AB - Treatment with 2 mM CuSO4 was used to induce a Drosophila melanogaster metallothionein (Mtn) promoter that had been cloned into a recombinant baculovirus. Careful study revealed that the Mtn promoter functioned as an inducible, if somewhat "leaky" promoter within the context of baculovirus infected cells. In the process of generating a recombinant-baculovirus, it was discovered that post-transfection treatment with copper resulted in a 10-fold increase in the production of recombinant virus. This effect on virus production was specific to transfection, as treatment of infected cells with copper did not increase the production of virus. Treatment of infected cells with copper did, however, extend the period of expression of the polyhedrin and p10 proteins by at least 12 h. These findings have practical applications for the production of recombinant baculoviruses and the subsequent expression of foreign proteins using baculovirus expression vectors. PMID- 9343701 TI - Studying electric field effects on embryonic myocytes. AB - Several problems arise when electrophysiological measurements are attempted on cells exposed to an electric field. In addition to field distortion produced by the reference electrode, membrane potential measurements by conventional microelectrode or patch-clamp techniques suffer serious interference from the applied field. We describe here a novel method for measurement of cardiac myocyte response to an alternating electric field that avoids these problems by sensing the mechanical activity of the cells rather than their electrical activity. A miniature electromechanical force transducer is used for this purpose. A glass pipet is attached to the force transducer, and only this pipet makes actual contact with the cell preparation. The resistive elements of the transducer are arranged as two legs of a Wheatstone bridge. Contractile activity of the cells produces small displacements of the micropipet and a resulting change in the transducer resistances. The Wheatstone bridge output is a current signal that is detected and converted to a voltage signal by a picoammeter before amplification and recording for later analysis. The technique may find applications in a variety of experimental studies of contractile tissues. PMID- 9343702 TI - Multiplex systems for the amplification of short tandem repeat loci: evaluation of laser fluorescence detection. AB - Short tandem repeat (STR) loci are ideal markers for personal identification and for genomic mapping. Two fluorescent multiplex systems, each designed for simultaneous PCR amplification of four polymorphic STR loci (HUMCSF1PO, HUMTPOX, HUMTH01 and HUMVWFA31, and HUMF13A01, HUMFESFPS, HUMBFXIII and HUMLIPOL), were evaluated on three laser fluorescence detection instruments. Concordant DNA typing results were obtained with all three detection methods. These fluorescent multiplex STR systems offer an accurate, reproducible and versatile method of DNA profiling that is well-suited for forensic identity testing and other genetic analyses. PMID- 9343703 TI - Temperature-induced expression of human-mouse chimeric Fab. AB - The temperature-induced expression vector pHZ01 with lambda PRPL promoter for the efficient expression of human-mouse chimeric Fab was constructed. Three kinds of chimeric Fab were expressed in E. coli: anti-prostate specific antigen (PSA) chimeric Fab, anti-lysozyme (HEL) chimeric Fab, and anti-tetanus toxoid (TT) chimeric Fab. All the soluble chimeric Fabs expressed showed specific antigen binding activities. PMID- 9343704 TI - The construction and expression of deletion mutant of human pro-urokinase cDNA. AB - The human pro-urokinase mutant deleting 150-156 amino acids was constructed by overlap-extension PCR and other molecular cloning techniques. The mutant was expressed transiently in COS-7 cells and constitutively in CHO cells; the expression level is 450-500 IU/10(6) cells/24 h. SDS-PAGE and Western blotting analysis showed that the molecular weight of the expression product is 54 kDa, which is similar to that of the mature pro-UK and recombinant pro-UK (rPro-UK) expressed by full-length cDNA. Most of the products exist in the form of single chain; the percentage of single chain is much higher than that of rPro-UK. In addition, the mutant product is more resistant to proteinases and its affinity to fibrin is also improved slightly. PMID- 9343705 TI - Cloning and sequence analysis of the 5'-flanking region of goat beta lactoglobulin gene. AB - We were the first to report 5'-flanking sequences of goat beta-lactoglobulin (beta LG) gene (867 bp) including a promoter (770 bp) and part of the first exon (97 bp). The fragment was isolated from Chinese goat tissue with PCR, and then cloned and sequenced. Sequence comparison showed 94.6% homology with sheep and 88% homology with cattle, respectively. There are highly conserved transcription factor binding sites as well. It suggested that the proximal 406 bp of 5' flanking sequences of sheep beta LG possess strong tissue-specific DNAaseI hypersensitive sites. Sheep beta LG transgenes containing only the 406-bp 5' flanking sequences were efficiently expressed in the mouse mammary gland at a high level. These goat beta LG 5'-flanking sequences could possibly direct heterologous protein expression in the milk of transgenic animals. PMID- 9343706 TI - The selective recognition of antibody IgY for digestive system cancers. AB - Biological methods for cancer therapies are very important. A small and efficient target carrier is the key component for anti-cancer drugs. In our laboratory, the antibody IgY was extracted from egg yolk of a SPF hen. The SPF hen was immunized with an antigene of P110 protein which was purified from human stomach cancer MGC 803 cells. Results indicated that the antibody IgY can specifically recognize gastrointestinal system cancers. It may become an important carrier for antitumorigenic drugs. PMID- 9343707 TI - Production of recombinant goldfish growth hormone I in a baculovirus expression system. AB - The cDNA sequence encoding the growth hormone (GH) I gene of goldfish (gf GH-I) was cloned into the pSXIVVI+ X3 baculovirus transfer vector. This transfer vector, without the initiation codon (ATG) and using a SynXIV promoter to activate transcription, was constructed for a baculovirus expression system. The recombinant transfer vector with the gf GH-I insert was used to produce the recombinant baculovirus TnNPV-SX+ gf GH-I 21a. Both in vivo and in vitro approaches were used to test the expression of growth hormone I gene using TnNPV SX+ gf GH-I 21a. For in vivo studies, larvae of Trichoplusia ni were infected with the recombinant baculovirus. Four days later, growth hormone-like immunoreactivity was detected in the hemolymph of these infected larvae. For in vitro studies, insect Spodoptera frugiperda 9(Sf9) cells were infected with TnNPV SX+ gf GH-I 21a. After incubation for at least 24 hours, growth hormone-like immunoreactivity was detected in Sf9 insect cells as well as in the culture medium. Western blotting analysis of larval hemolymph and Sf9 cell contents after viral infection revealed a protein band of 22.5 kDa immunoreactive to goldfish growth hormone antiserum. This is consistent with the predicted molecular weight deduced from the cDNA of goldfish growth hormone I gene. These results, taken together, suggest that the baculovirus expression system can be used to produce the recombinant growth hormone of a fish species. PMID- 9343709 TI - Fed-batch culture strategy for high yield of baker's yeast with high fermentative activity. AB - Based on the determination of the operational conditions for batch culture and fed-batch culture of baker's yeast by process analysis and continuous culture, a correlation, which describes the relationship between fermentative activity (FA) and specific growth rate (mu), was obtained. Combining this correlation and the exponential fed-batch culture equation, a fed-batch culture strategy was developed to obtain high yield and high fermentative activity by controlling mu at different stages. The results showed that when the initial and residual sugar concentrations were controlled to be 15-30 g/L and 3-6 g/L, respectively, different stirring speeds were applied at different stages to provide optimal oxygen transfer conditions. When this proposed fed-batch culture strategy was used in a fed-batch culture, the yield of baker's yeast reached 0.432 g/g with high fermentative activity (1180 ml). Thus, the combined bed-batch culture of baker's yeast with high yield and high fermentative activity was realized. PMID- 9343708 TI - Expression of HBsAg gene in transgenic goats under direction of bovine alpha-S1 casein control sequence. AB - An expressive construct combining hepatitis B surface antigen gene and bovine alpha-S1 casein control sequence (a 16-kb fragment), lambda 106, was made successfully using DNA recombination technology. The construct was then introduced into fertilized goat eggs with a transgenic technique. As the eggs developed into adults which, in turn, began to secrete milk after parturition, ELISA assay demonstrated HBsAg in the milk, thus showing successful expression of the construct in the mammary gland of the G0 transgenic goats and the secretion of the expression product into the milk. PMID- 9343710 TI - Optical resolution of DL-alanine by using immobilized Aspergillus oryzae cells. AB - Mycelium pellets with diameters of 1-2 mm and containing abundant aminoacylase were obtained by the liquid fermentation of Aspergillus oryzae 3042. By the cross linking method with reagents of gelatin and formaldehyde, the immobilized A. oryzae cells (IAC) were prepared with much higher activity and reactive properties. The effects of factors on enzymatic activity of IAC were investigated. When substrate concentration was less than 0.15 mol/L, the Michaelis-Menten mechanism was suitable for this reaction. PMID- 9343711 TI - Study on enzyme electrode biosensor of choline. AB - Choline oxidase was immobilized at a hydrogen peroxide electrode and the enzyme electrode was used for the amperometric determination of choline. The linear range is 0-200 mg/L with a response time of 40 seconds and a 25-microliter sample injection. The relative standard deviation (RSD) is less than 1.5% in 20 assays. The enzyme membrane can be used continuously at 25 degrees C for 60 days. The recovery rate of this method is 100.3-102.3%. PMID- 9343712 TI - Long-term studies on patients with absence and bilateral spike-wave complexes: 430 patients, up to 52 years follow-up. AB - This study involved long-term changes, up to 52 yrs, in 430 patients with absence (A) attacks and bilateral spike and wave complexes (BSW). Nearly 2/3 had additional GTC attacks and only about 1/4 had A alone. Atonic attacks (7%), complex partial (CP) (4%) and other types of seizures also occurred in the group. Distinctive features of the GTC group were diffuse slow waves and 6/sec spike and wave complexes, and of the A alone were BSW as the only abnormality and later normal records. For CP or focal motor (FM) attacks, nearly all had focal spikes and these seizures appeared between 15-25 yrs of age. The background alpha rhythm with increasing age showed more rhythmicity, but lower amplitudes in all of these patients. Onset age was an important factor in that the highest peak was at 6 yrs, mainly from those with A only and a minor peak appeared at 13-14 yrs, mainly from the patients with GTC seizures. One major difference between the group with A alone and those with GTC attacks was that normal records developing with A alone usually remained normal, while those with GTC attacks usually became abnormal again. The 6/sec spike and wave complexes usually appeared after the onset of GTC attacks, often at 18-19 yrs of age. Diffuse slowing usually appeared after GTC attacks at around 16 yrs after the onset of these seizures. Possible forerunners of BSW were multifocal spikes at < 4 yrs of age, occipital or temporal spikes at 5-16 yrs and temporal spikes after 16 yrs of age. CP seizures could be predicted from the EEG by focal spikes, especially on the temporal lobe, appearing over long periods of time in records with an inconsistent BSW. Patients with A extending into adulthood were distinctive by a later onset of A and a history of GTC attacks. In general, more females were seen in this study but there were more males with A alone and more developed a normal record. Females often had a later onset of A, with GTC attacks, and the older the patient in this study the more likely the patient was female. Evidence is seen in this study for prediction of (1) future BSW by different types of focal spikes at different ages, (2) CP (or FM) seizures by focal spikes and an inconsistent BSW, (3) GTC seizures by onset age, female sex, and EEG reverting to an abnormal record after normalizing, (4) A attacks alone by onset age, male sex and finally a normalized EEG, and (5) GTC or myoclonic attacks by the presence of BSW as the only abnormality. PMID- 9343713 TI - Excessive daytime sleepiness associated with idiopathic alterations of consciousness. AB - Sleep/wake patterns were recorded by continuous 24-hour ambulatory polysomnography in 339 patients, who had episodes of altered consciousness. Patients were recorded while they were outside the hospital. From a seven-channel montage of electrodes affixed below the hairline, sleep polygraphic EEG was easily read from T3-T4, EOG from F2-F8 and EMG from T3-T6. Sleep was staged by analysis of aural signals on 60 times real time playback, augmented by continuous visual display and selected frozen frames. Patient major sleep period patterns reflected those reported for general populations. Unexpectedly, 47% of the patients took daytime naps and 44% of the nappers took more than one nap. Naps had a mean duration of 71 minutes. Those who took no naps slept significantly longer at night by 23 min. Napping reduced night sleep much more in patients who did not take CNS-acting medications. We conclude that excessive sleepiness may in part explain complaints of episodically altered consciousness. PMID- 9343714 TI - Leigh syndrome, cytochrome C oxidase deficiency and hypsarrhythmia with infantile spasms. AB - A rare patient with infantile spasms, hypsarrhythmia, cytochrome c oxidase deficiency and Leigh syndrome is reported. Although rare, infantile spasms and Leigh syndrome may occur simultaneously. Leigh syndrome should be included in the differential diagnosis of infantile spasms. PMID- 9343715 TI - Single-trial analysis of P3 in patients with generalized epilepsy. AB - The latencies and amplitudes of averaged P3, and the latencies, amplitudes and frequency components of single EEG responses to target tones were analyzed in 9 control subjects (CS group), 6 epileptics whose mean IQ was 100 (EP group) and 6 epileptics whose mean IQ was 52 (RE group), using an auditory oddball task. All of the subjects responded to the target tones correctly and there were no differences in the incidence of error in response to the target tones, or in the latencies and amplitudes of the averaged P3 among the three groups. However, the reaction times (RTs) in the RE group were significantly longer than those in the other groups (P < 0.05). Single EEG responses to target tone (single-trial ERPs) were classified into 2 types, those with and those without the P3 component. Type 1 had the P3 component and was observed in 42% of all of the responses in the RE group, significantly less than those in the CS (64%) and EP (61%) groups. The peak latencies of P3 in type 1 were similar among the three groups, but the amplitudes of P3 in type 1 in the RE group were significantly greater than those in the CS and EP groups. RTs in the RE group were significantly longer than those in the other groups, and had no correlation with the P3 latencies of type 1. There was little difference in the results of the frequency analysis among the three groups. These results suggest that all subjects in three groups recognized the target tones correctly, but they did not evaluate every target tone, since the incidence of P3 was almost 60% in the CS and EP groups, and 40% in the RE group. The characteristics of cognition and evaluation in three groups were the same, but the decrease in incidence of evaluation and the dissociation between the cognition and the response execution might be caused by impairment of the subject-environment contact mechanism, which resulted in the decrement of IQ in the RE group. PMID- 9343717 TI - Clinical EEG findings in mania. AB - Clinical EEG findings from 202 hospitalized manic patients repeated during 131 recurrences of mania were described. Results were considered in the light of current issues in the literature including the incidence of EEG abnormalities and minor variations, relationships between EEG and family history, EEG lateralization and longitudinal course of illness. The majority of patients had normal EEGs or mild nonspecific deviations compatible with effects of psychoactive medications. More definitive EEG abnormalities were observed in 16 percent. Microsleep occurred in 19 percent and small sharp spikes were found in 17 percent of those who drowsed, with lower incidences of 14 and 6 positive bursts and 6 Hz spike-and-slow-waves. Significant relationships between moderate or severe EEG abnormalities and negative familial loading were identified. Lateralized EEG abnormalities appeared in 9 percent of cases, involving the left side significantly more often than the right. With one exception EEG recordings during subsequent episodes did not suggest structural brain changes. Clinical EEG studies are useful in discriminating between primary and secondary affective disorders. They are also sensitive to effects of lithium and other psychoactive medications. The significance of EEG variations including microsleep and other atypical features continues to be elusive. Issues relating to heritability, hemispheric dysfunction and longitudinal course of illness merit further investigation. PMID- 9343718 TI - Aging, smoking and EEG coherence: a preliminary study. AB - Although the cigarette smoking habit is prevalent in young, middle aged and elderly adults, it is yet unknown whether a long term smoking history alters the aging brain and/or whether the aging brain demonstrates an altered sensitivity to acute smoking. Inter- and intrahemispheric EEG coherence was compared in 20 young (18-39 years) adults (10 smokers, 10 nonsmokers) and 20 elderly (64-81 years) adults (10 smokers, 10 nonsmokers). The acute effects of sham inhaling on a nonlighted cigarette and cigarette smoking on EEG coherence was also compared in young and elderly adult smokers. In general, elderly adults exhibited reduced interhemispheric coherence values relative to young adults and, depending on the frequency band, age effects varied with recording site. Smokers of both age groups exhibited greater interhemispheric total alpha coherence values than nonsmokers. Similar smoker status effects were found with fast alpha but this varied with recording site. Relative to sham smoking, acute cigarette smoking reduced interhemispheric slow alpha in both young and elderly smokers but reduced total alpha coherence only in elderly smokers. The results are discussed in relation to normal and pathological aging, including dementia. PMID- 9343716 TI - Epilepsy: a costly misdiagnosis. AB - Nonepileptic events misdiagnosed as manifestations of epilepsy are a significant problem. Five representative cases suggest that a diagnosis of epilepsy should be based on a careful history and analysis of the clinical event. The clinician should make a diagnosis of epilepsy only when there is definite positive support and not on the basis of poorly characterized paroxysmal episodes and a few minor and nonspecific EEG findings. Effective treatment presupposes correct diagnosis and early diagnosis appears to be correlated with better prognosis. The annual cost of nonepileptic spells misdiagnosed as epileptic can be estimated at between $650,000,000 and $4,000,000,000. Aggressive efforts to establish a definite diagnosis of spells not clearly epileptic is both medically and economically indicated. PMID- 9343719 TI - Movement-related cortical potentials and contingent negative variation in olivopontocerebellar atrophy. AB - Movement-related cortical potentials (MRCPs) and contingent negative variation (CNV) were recorded in 8 cases of olivopontocerebellar atrophy (OPCA) and 8 age matched healthy controls. The amplitude of the negative slope (NS') was smaller in the OPCA group than in the healthy control group. The amplitude of late CNV was smaller in OPCA group than in the healthy control group. These abnormalities in MRCPs and CNV were improved by intravenous infusion of thyrotropin releasing hormone. PMID- 9343720 TI - Painful knee joint prostheses: evaluation for loosening by combined radionuclide arthrography and transmission imaging. AB - Interpretation of radionuclide arthrograms (RNA) of the knee after total knee joint replacement (TKR) may be difficult because of the lack of sufficient anatomic reference information. Additional transmission images may provide the necessary reference information required for correct interpretation of RNA of the knee. METHODS: Tc-99m tin colloid (20 MBq) RNA was combined with cobalt-57 transmission imaging in six patients with painful TKRs in whom loosening of the TKR was suspected. Knee surgery was then performed on these six cases. RESULTS: RNA correctly detected loosening of the tibial component in five of six cases, but did not detect loosening of the femoral component in any of three cases. CONCLUSION: RNA combined with transmission imaging is recommended for detection of loosening of the tibial component of a TKR. RNA may not be of value for detecting loosening of the femoral component. PMID- 9343721 TI - A prospective comparison of high-resolution planar, pinhole, and triple-detector SPECT for the detection of renal cortical defects. AB - To compare the detection rate of renal cortical defects with Tc-99m dimercaptosuccinic acid (DMSA) using triple-detector SPECT, pinhole, and planar cortical scintigraphy, the authors prospectively studied 80 kidneys in 40 patients (26 males, 14 females) who ranged in age from 3 months to 26 years (mean: 7.5 years). They found single or multiple definite defects in 30 kidneys using SPECT, 23 using pinhole imaging, and 17 using planar imaging (McNemar's test, two-tailed, P < 0.001 and P = 0.03, respectively). SPECT was significantly better than pinhole imaging at demonstrating definite defects (P = 0.008). This study indicates that SPECT, and to a lesser extent pinhole, are superior to planar imaging for conclusively demonstrating renal cortical defects. PMID- 9343722 TI - Technetium-99m HMPAO labeled leukocytes as the primary diagnostic tool in a case of brain abscess. AB - The authors present a case in which Tc-99m HMPAO labeled autologous leukocytes were used to demonstrate a brain abscess in a patient undergoing evaluation for fever of unknown origin. The abscess was demonstrated on both 1-hour and 24-hour images. The positive 1-hour image led to CT and MRI studies, which are included for correlation. In addition to its previously identified role as a secondary diagnostic test in the differentiation of tumor and abscess, the authors propose that Tc-99m HMPAO is useful as a primary diagnostic tool in the identification of brain abscess. Furthermore, the authors suggest that Tc-99m HMPAO is preferable to In-111 labeled leukocytes because of its better resolution and earlier imaging characteristics. PMID- 9343723 TI - Radionuclide imaging in emphysema after lung volume reduction surgery. AB - Three dimensional (3D) surface displays of dynamic Xe-133 and Tc-99m MAA SPECT were performed to evaluate regional lung function in two patients with pulmonary emphysema before and after thoracoscopic lung volume reduction surgery. The 3D displays were reconstructed using a color-illuminated, surface-rendering technique. For the Xe-133 studies, a fusion display of 3D equilibrium (EQ) and 3 minute washout (WO) images was used to show the distribution of Xe-133 retention, which were transparently seen within the entire lung contours delineated by the EQ image. In both patients, these 3D displays allowed an overview of the localized poorly functioning lungs with Xe-133 retention or reduced perfusion with geometric realism. The location and extent were more easily comprehended on the displays compared to the slice-by-slice presentations of tomograms. Postoperatively, the displays efficiently revealed restored function in the remaining lungs. The 3D surface displays of the two SPECT procedures providing topographic information of regional lung function appears to contribute to the treatment strategy of this surgery. PMID- 9343724 TI - Metastatic pulmonary pheochromocytomas: positive I-123 MIBG SPECT with negative I 131 MIBG and equivocal I-123 MIBG planar imaging. AB - Pulmonary metastases from malignant pheochromocytoma were seen with I-123 MIBG SPECT in a 16-year-old girl but were not visualized with I-123 MIBG planar imaging. She had a left adrenalectomy for a pheochromocytoma 7 years earlier. Two small pulmonary nodules were seen on chest X-ray and CT scans. PMID- 9343725 TI - Multiple brown tumors in parathyroid carcinoma mimicking metastatic bone disease. AB - An unusual case of multiple brown tumors due to parathyroid carcinoma is reported. The patient presented with lower leg pain. Plain radiographs demonstrated multiple lytic lesions of the lower legs and a Tc-99m MDP bone scan depicted multiple areas of increased uptake suggesting skeletal metastases. Tc 99m sestamibi tumor scintigraphy showed multiple sites of tumor uptake in bones and a large area of increased uptake with a cystic component in the right lower pole of the thyroid gland. An open biopsy from the right tibial lesion revealed a brown tumor. A large parathyroid carcinoma with a necrotic cyst was removed. After parathyroidectomy and right thyroid lobectomy, the patient became free of bone pain and serum PTH levels normalized. A 9-month follow-up Tc-99m MDP bone scan demonstrated less intense uptake in the pelvis, tibia, and fibulae. Nine month follow-up tumor imaging with Tc-99m MIBI revealed disappearance of the preoperative uptake of multiple brown tumor. PMID- 9343726 TI - Intestinal lymphangiectasia: value of Tc-99m dextran lymphoscintigraphy. AB - Intestinal lymphangiectasia is a common cause of protein-losing enteropathy characterized by diarrhea, generalized edema, enteric protein loss, hypoproteinemia, and lymphopenia. Diagnosis is based on demonstration of enteric protein loss and characteristic small bowel mucosal histology. Various imaging modalities including barium studies, computed tomography, and lymphangiography have had limited clinical use. The authors report a case of intestinal lymphangiectasia in which Tc-99m dextran lymphoscintigraphy played a significant role in the patient management. PMID- 9343727 TI - Improved cardiac iodine-123 metaiodobenzylguanidine accumulation after drug therapy in a patient with Parkinson's disease. AB - A 72-year-old woman with Parkinson's disease and autonomic dysfunction underwent I-123 MIBG cardiac imaging to study sympathetic neuronal integrity. This revealed a defect in the infero-posterior wall. Tc-99m sestamibi myocardial perfusion imaging revealed no abnormalities. On follow-up I-123 MIBG SPECT, the defect was less prominent and the parkinsonian symptoms were ameliorated by drug therapy. Cardiac I-123 MIBG imaging may be a promising new method for evaluating drug therapy in patients with Parkinson's disease. PMID- 9343728 TI - Relative sensitivity of Tc-99m WBC versus In-111 WBC in a patient with Crohn disease and steroid use. AB - Numerous studies have shown that chemotaxis is affected by certain antibiotics and steroids. The authors present the case of a patient with Crohn disease relapse with multiple small-bowel fistulae and mesenteric abscesses. Whereas the Tc-99m WBC scan failed to show the intra-abdominal inflammatory foci, an In-111 WBC scan performed within a week delineated the abscesses very well, and these were later confirmed at surgery. This case is presented not only to illustrate the relative sensitivities of a Tc-99m WBC versus an In-111 WBC scan, but also to discuss the impediment to polymorphonuclear chemotaxis by steroids, which may be a contributory factor to the sensitivities of the different radiopharmaceuticals selected for detection of intra-abdominal septic foci. PMID- 9343729 TI - Extensive Ga-67 accumulation in lymph nodes and cutaneous metastases in gastric cancer. PMID- 9343730 TI - Tc-99m labeled antigranulocyte monoclonal antibody imaging for the detection of an abdominal abscess. PMID- 9343731 TI - Lumbopleural cerebrospinal fluid shunt assessment with Tc-99m DTPA scintigraphy. PMID- 9343732 TI - Bowel compression on the gallbladder mimicking a gallstone on cholescintigraphy. PMID- 9343733 TI - Forearm splints seen on bone scan in a weightlifter. PMID- 9343734 TI - False negative Tc-99m hepatobiliary scan in a patient with Osler-Weber-Rendu disease. PMID- 9343735 TI - Detection of a small gastrinoma by combined radiologic and scintigraphic techniques. PMID- 9343736 TI - Extensive extraskeletal osteosarcoma in the liver and abdomen as demonstrated by bone scintigraphy. PMID- 9343737 TI - Bone scintigraphy in an uncommon presentation of metastatic lung carcinoma. PMID- 9343738 TI - Leukemic infiltration mimicking epididymo-orchitis on scrotal scintigraphy. PMID- 9343740 TI - Malrotation of the gastrointestinal tract mimicking a bile leak. PMID- 9343739 TI - Marrow visualization on an abdominal blood pool scan. PMID- 9343741 TI - Diagnostic dilemma: lymphocele with features of a urinary leak. PMID- 9343742 TI - Scintigraphy of proximal femoral focal deficiency. PMID- 9343743 TI - Bladder displaced by stool-distended rectosigmoid colon presenting on bone scan as a pelvic soft tissue mass. PMID- 9343744 TI - Current readings in nuclear medicine. PMID- 9343746 TI - Cytoprotective activity in the gastric mucosa of rats exposed to carbon tetrachloride-induced liver injury. AB - The present study was undertaken to evaluate the cytoprotective activity in the gastric mucosa of rats subjected to CCl4-induced liver injury. Response of gastric mucosa to absolute ethanol insult or acid (pylorus ligation) after CCl4 challenge was analyzed. Intraperitoneal administration of CCl4 increased plasma AST and ALT, but liver protein and glycogen levels were decreased; in addition, gastric acid secretion was significantly increased with respect to control animals (1541 +/- 266 vs. 629 +/- 25 mu eq H+; p < 0.001). Microscopical gastric erosions were observed in 3/10 animals after CCl4 challenge. Pylorus-ligated as well as CCl4-challenged rats developed increased susceptibility to gastric lesions, compared to control (lesion indices: 4.6 +/- 0.20 vs 2.8 +/- 0.13; p < 0.05), while showing increased resistance to absolute ethanol-induced gastric damage (30.4 +/- 11.2 vs 89.7 +/- 9.7 mm, p < 0.01). PGE2 levels in the gastric mucosa were not influenced by exposure to CCl4. Ultrastructural studies revealed the presence of continuous ethanol-resistant and apparently more adherent layer of mucus in CCl4-challenged animals. Morphological evaluation revealed an increase in Alcian blue-stained mucus. A dual condition of enhanced sensitivity to HCl with increased tolerance to ethanol was observed in gastric mucosa of CCl4 treated animals. These observations could be explained by the amount and/or composition of protective mucus layer in the gastric mucosa. PMID- 9343745 TI - Arachidonic acid induces DNA-fragmentation in human polymorphonuclear neutrophil granulocytes. AB - We analyzed DNA-fragmentation in human polymorphonuclear neutrophil granulocytes (PMNs) from healthy donors after addition of exogenous arachidonic acid (AA) and eicosapentaenoic acid (EPA) by flow cytometry (propidium iodide staining of DNA and DNA strand break detection). The PMNs were incubated from 30 min up to 48 hours in RPMI 1640 which was supplemented with different concentrations of AA or EPA (5-40 microM). In contrast to EPA the addition of AA led to a significant increase in apoptosis up to 67.8% compared to the RPMI-control whereas the addition of EPA led to an inhibition of DNA-fragmentation. When the cells were incubated with MK 886 (1 microM, inhibitor of leukotriene biosynthesis) an increase in DNA-fragmentation (up to 63.3%) was observed. Conversely, in the presence of indomethacin (1 microM, inhibitor of prostanoid synthesis) an inhibition in DNA-fragmentation (up to 60.9%) occurred. Furthermore, preincubation of PMNs with pentoxifylline (1mM, phosphodiesterase inhibitor) reduced AA-stimulated DNA-fragmentation up to 43.4%. These data provide evidence for the involvement of AA and distinct AA metabolites in the regulation of apoptosis in human PMNs. PMID- 9343747 TI - Retrograde migration of starch in the genital tract of rabbits. AB - This study in a rabbit model simulates contamination with glove powder in association with a routine gynaecological examination. Large individual variations of powder contamination were found and there were no overall statistically significant differences between the control and experimental animals. The findings are supported by the observation that some but not all women develop adhesions after gynaecological surgery. Analyzes of variances indicate differences in the migration of starch particles in the genital tract with the highest amount of particles found three days after starch contamination of the vagina. Since no adhesions were observed, there would probably need to be an ongoing post surgical or post infectious inflammation in the tissue, when the starch particles are added. Starch powder from latex gloves can cause adhesions and increase the risk of latex allergy in healthcare workers. Retrograde migration in the genital tract cannot be excluded, powdered examination products should be eliminated from the gynecologicla examination room. PMID- 9343748 TI - Cytokine responses of human blood monocytes stimulated with Igs. AB - Using an in vitro system for stimulating human peripheral blood mononuclear cells (PBMC) with immobilized Ig, patterns of cytokine production as a function of different Ig classes and subclasses were elucidated. Wells were coated with IgA, IgG1, IgG2, IgG3 or IgG4. Equivalent protein content on surfaces of wells was demonstrated by a human kappa chain ELISA. Isolated human PBMC were added to Ig coated wells and incubated for 24 hrs before supernatants were assayed for cytokines. The IgG subclasses showed differences in cytokine production stimulated from PBMC, with the relative stimulation for TNF alpha being IgG2 > or = IgG3 > or = IgG1 > IgG4 and for IL-6 production, IgG2 > or = IgG3 > IgG1 = IgG4. In contrast, the relative stimulation for IL-8 was IgG1 = IgG2 = IgG3 = IgG4. IgA caused less production of TNF alpha when compared to IgG2, but similar levels of IL-8. Such differences may have important implications in the pathogenesis of immune complex mediated diseases. PMID- 9343750 TI - A rise in ionized calcium activates the neutrophil NADPH-oxidase but is not sufficient to directly translocate cytosolic p47phox or p67phox to b cytochrome containing membranes. AB - Neutrophil production of reactive oxygen species is dependent on an assembly process that involves a translocation of the cytosolic NADPH-oxidase components (p47phox; p67phox; Rac2) to a b cytochrome containing membrane. Based on the fact that an intracellular Ca2+ rise can activate the oxidase without any extracellular release of reactive oxygen species, we suggest that the oxidase can be assembled in a membrane distinct from the plasma membrane. Disintegrated cells were used to monitor Ca2+ dependent membrane binding of neutrophil cytosolic proteins. Membranes containing the b cytochrome part of the oxidase, i.e., specific granules and plasma membranes/secretory vesicles, were used in the translocation experiments. Several cytosolic proteins were found to translocate to specific granules as well as the plasma membranes/secretory vesicles, one of them being annexin I. Using antibodies in the blotting assay against the cytosolic oxidase components p47phox and p67phox, we could show that no Ca2+ dependent translocation of these cytosolic proteins occur to neither of the b cytochrome containing membranes. PMID- 9343749 TI - Study on paradoxical effects of NSAIDs on platelet activation. AB - We recently described a stimulatory effect of high doses (> 100 mumol/L) diclofenac on platelet adhesion. In this study we extend our research to the possible biochemical mechanisms of the observed effects, to other non steroidal anti-inflammatory drugs (NSAIDs) (flurbiprofen, indomethacin, acetylsalicylic acid, ibuprofen, nitrofenac and nitroflurbiprofen) and to the effect of high doses diclofenac and flurbiprofen on platelet aggregation. We observed that high doses of diclofenac and of flurbiprofen, but not of the other tested NSAIDs, increased platelet adhesion at doses ranging from 100 to 500 mumol/L, an effect completely removed by the 12-lipoxygenase-inhibitor nordihydroguaiaretic acid. Moreover, they had no pro-aggregating effect, inhibiting platelet aggregation induced by 10 mumol/L arachidonic acid and dose-dependently increasing the [Ca2+]i. Finally, whereas no basal nitric oxide release by washed platelets was detected, when platelets were incubated by 500 mumol/L diclofenac or flurbiprofen, the production of nitric oxide, as measured by amounts of nitrite released, was 4.4 +/- 0.5 and 3.8 +/- 0.4 pmol/5 x 10(8) platelets/min, respectively. Our data indicate that high doses diclofenac and flurbiprofen are promoters of the early phases of platelet activation, probably through the 12 lipoxygenase pathway. PMID- 9343751 TI - Staphylococcal culture supernates stimulate human phagocytes. AB - Phagocytes play a major role in host defense against staphylococci as well as in the pathophysiology of Gram-positive septic shock. In Gram negative sepsis, the main mediator, LPS exerts its effects as easily suspendable mediator. In Gram positive sepsis the main mediator is still not found, therefore we studied the interaction of soluble staphylococcal products with phagocytes. Staphylococcus aureus supernates (SaS) were harvested from several laboratory and clinical strains that were grown to late-log phase. These supernates upregulated CD11b/CD18 expression on human neutrophils even in a 100-fold dilution. SaS also induced the release of TNF-alpha and IL-1 beta by human monocytes. Control experiments excluded peptidoglycan, lipoteichoic acid, alpha and delta toxin, leucocidin, TSST-1 and all enterotoxins as sole mediators. Endotoxin contamination was also excluded. SaS was heat-stable; incubation for 45 minutes at 100 degrees C did not affect its activity. Compared to purified peptidoglycan and intact bacteria per bacterium, SaS had a higher potency in stimulating phagocytes. We hypothesize that there are more--yet unknown--soluble staphylococcal products which are very important in phagocyte stimulation. PMID- 9343752 TI - Validating quality indicators for hospital care. AB - BACKGROUND: Many of the indicators used to monitor the quality of hospital care are resource intensive and ineffective. Furthermore, current efforts to develop new indicators for report cards are generally directed at the evaluation of health plans and are not constructed to help providers (physician groups, hospitals, and health plans that contract to provide care to patients) find and fix problems with the quality of care at their organizations. FOUR QUESTIONS: Before using an indicator, four questions should be posed: (1) When cases identified by the indicator are examined, can one find a set of definable and preventable processes of care known to lead to a bad outcome? (2) Can a review instrument be created that will allow providers to identify which process problems are present? (3) Are there substantially more process problems in those cases identified by the indicator than in those cases not identified by the indicator, and can the sensitivity and specificity of the indicator be defined? and (4) Is the indicator primarily useful for quality improvement efforts by a provider, or is it also useful as an external measure of quality across providers? A FOUR-STEP FRAMEWORK: Four corresponding steps comprise an efficient validation method to produce indicators that detect deficiencies in an important process-outcome continuum, help produce the tools to find the deficiencies, document the efficiency of using the indicator to search for process problems, and define the appropriate use of the indicator. Use of such validated indicators, and the information about their utility, would allow providers to optimize the impact of money spent on quality improvement efforts. PMID- 9343753 TI - Pursuing clinical and operational improvement in an academic medical center. AB - BACKGROUND: An academic medical center in an increasingly competitive market, the University of California-Davis Medical Center in Sacramento started working with a consulting firm in 1995 to reduce overall operational costs and costs for the clinical processes involved in treating patients with specific conditions. ESTABLISHING THE TEAMS: Twelve operational efficiency (OE) teams and five clinical teams were commissioned, with a combined total of nearly one-half of the target cost reduction. The second wave of six clinical teams was simultaneously initiated in late spring 1996. THE IMPROVEMENT METHOD: The quality improvement process for clinical improvement teams included the review and inquiry method, which enables many pilot experiments to be conducted in parallel by work groups and coordinated by the main task team. RESULTS AND CASE STUDIES: Within six weeks of launching, the 12 OE teams achieved their goals and identified savings opportunities of more than $27 million. One OE team, medical records, had set a goal of $514,000 in cost reduction for a three-year period and achieved the first year goal of $190,000. For a clinical team on interventional cardiology, the clinical benchmark data revealed that the cost per case of providing cardiac catheterization was greater than for all three benchmark groups. These patients, including 270 patients per year, showed a possible savings through process improvement of nearly $1.4 million. From January 1996 through March 1997, the rate of occurrence of complications decreased from 5.5% to 3%. EPILOGUE: Physicians gradually accepted more responsibility and accountability for controlling and reducing costs, while maintaining their traditional role as advocates for improved patient care. PMID- 9343755 TI - The quality of dying: how can we improve care at the end of life? An interview with the IOM's Marilyn Field. Interview by Steven Berman. PMID- 9343754 TI - Improving the processes of care and outcomes in obstetrics/gynecology. AB - BACKGROUND: The obstetrics/gynecology department of York Hospital (York Health System, York, Pennsylvania) initiated a program to improve the processes of care and control costs for common women's and newborns' health care services. Twelve clinical policies were established between June 1993 and February 1995. CONDUCTING THE QUALITY IMPROVEMENT (QI) PROJECTS: Using the plan-do-check-act (PDCA) improvement cycle method, the QI group established clinical pathways for high-volume conditions or procedures known to have low rates of complications and clinical guidelines for those conditions or procedures not requiring coordinated efforts of a group of health care professionals. EXAMPLE--PYELONEPHRITIS IN PREGNANCY: The literature had indicated that the prevalence of pyelonephritis can be decreased by identifying and treating asymptomatic bacteriuria early in prenatal care. After the validity of the clinical policy was demonstrated in the resident service, the policy was extended to all private obstetric practices. Dissemination of the finding that most of the admissions for pyelonephritis were for referred patients (for whom we had no control over prenatal care) or for patients referred by private physicians who were not yet following the guidelines quickly led to complete compliance by our obstetricians and other health care providers referring patients to the York Health System. RESULTS: The 12 clinical policies resulted in the elimination of 113 admissions and 5,595 inpatient days and in the reduction of the cost of patient care by $1,306,214 for the years 1994 1995 and 1995-1996 combined, without apparent adverse effects on patient health. CONCLUSION: A voluntary clinical policies program can change the culture of a department and lead to cost-effectiveness and better quality of patient care. PMID- 9343756 TI - Risks of noise-induced hearing loss for physical education teachers. PMID- 9343758 TI - Length of disability and cost of workers' compensation low back pain claims. AB - Although information exists on the cost of workers' compensation low back pain (LBP), there is limited information on the duration of lost work time as well as the association between cost and duration. For this study, cost and duration of lost work time information were derived from a large workers' compensation company's database for 1992 LBP claims (n = 106,961). The distribution of cost was skewed, with an average cost of a claim being 20 times higher than its median. A disproportionately small percentage of the costliest LBP claims (10%) were responsible for a large percentage of the total cost (86%). The distribution of length of disability (LOD) was also skewed, with an average of 102 days and a median of zero. The average and median LOD for those claims with at least one day of compensable disability was 303 and 39 days, respectively. As a "rule of thumb," it was found that of those claimants who remain on disability at the end of n weeks, approximately 50% will be off disability at the end of 6.n weeks. Additionally, the 7% of the claims with an LOD greater than one year accounted for 75.1% of the cost and 84.2% of the total disability days. Disability days that were accrued after one year of disability accounted for 59.3% of the total number of disability days. This result suggests that other LOD estimation techniques, which may not account for disability days beyond one calendar year (e.g., the Bureau of Labor Statistics Annual Survey of Occupational Injuries and Illnesses), may result in a marked underestimation of LOD. PMID- 9343757 TI - Solvent exposure in the railroad industry. PMID- 9343759 TI - Analysis of nicotine and cotinine in the hair of hospitality workers exposed to environmental tobacco smoke. AB - The purpose of this study was to determine if hair nicotine and cotinine levels reflect relative exposure to environmental tobacco smoke (ETS) in subjects who worked in the hospitality industry, where public smoking was permitted. Hair samples from 26 subjects were analyzed by gas chromatograph/mass spectrometry techniques for nicotine and cotinine. An exposure gradient was shown for nicotine but not cotinine. Among nonsmokers, those working in bars where there are no public smoking restrictions had the highest hair nicotine levels, which were close to levels found in smokers. Nicotine measured in hair is useful as a biological marker for exposure to ETS from multiple sources. Bar workers in particular are exposed to high levels of ETS, which may adversely affect the health of nonsmokers. PMID- 9343760 TI - Vitamin B6, vitamin C, and carpal tunnel syndrome. A cross-sectional study of 441 adults. AB - As part of an ongoing study of carpal tunnel syndrome (CTS) in industry, we measured plasma concentrations of pyridoxal 5'-phosphate (PLP, a measure of vitamin B6 status) and total ascorbate (ASC, a measure of vitamin C status) in 441 adult volunteers from six industries and a university exercise study. In the entire study group and in non-vitamin users (n = 218), there were no significant differences in mean plasma PLP or ASC concentrations between controls (neither symptoms nor slowing), subjects with symptoms only, subjects with median nerve slowing only, or subjects with CTS (symptoms + slowing). In male non-vitamin users (n = 137), there were significant inverse univariate associations between plasma PLP concentration and the prevalence of pain, the frequency of tingling and nocturnal awakening, and the Phalen test result. In this same subgroup, the ASC/PLP ratio was directly associated with the prevalence of pain and nocturnal awakening, and with the frequency of pain, tingling, and nocturnal awakening. In multivariate analyses, plasma ASC concentration predicted more median nerve slowing and confirmed CTS, and vitamin or vitamin interaction variables were independent predictors of 20 CTS-related outcomes. These multivariate relationships often occurred only after adjustment for age, gender, body mass index, serum alkaline phosphatase activity, or tobacco use. We conclude that there are significant relationships between plasma vitamin levels and both components of CTS (specific symptoms and median nerve slowing). The interaction between plasma PLP and ASC appears to be particularly important with respect to symptoms. PMID- 9343761 TI - The prevalence of pulmonary and upper respiratory tract symptoms and spirometric test findings among newspaper pressroom workers exposed to solvents. AB - To investigate the relationship between exposure to organic solvents and the presence of pulmonary and upper respiratory tract mucous membrane symptoms, we conducted a cross-sectional study of 215 newspaper pressroom workers who were occupationally exposed to organic solvent and lubricant mixtures. Thirty-four compositors, who were not occupationally exposed to the solvents or lubricants, served as controls. Pressroom workers and compositors underwent spirometric testing and were also asked about the presence of cough, phlegm, hemoptysis, dyspnea, wheezing, chest tightness, nose or throat irritation, eye irritation, and sinus trouble. The spirometric results did not significantly differ between the two groups. However, the pressroom workers were significantly more likely to report pulmonary or upper respiratory tract mucous membrane symptoms than were compositors (P < 0.005). An exposure-response relationship could be demonstrated when comparing the number of solvents exposed with the total number of symptoms (P < 0.001). Similarly, an exposure-response relationship could be demonstrated when comparing the frequency of use of each of the seven solvents with the total number of symptoms (P < 0.002). Each of these findings was supported in a multivariable linear regression model that adjusted for potential confounders such as age, smoking history, and number of years in the industry. A high prevalence of these symptoms was reported even though the degree of exposure to solvents and lubricants was within the current permissible exposure limits. PMID- 9343762 TI - Mortality experience of a young petrochemical industry cohort. 1979-1992 follow up study of US-based employees. AB - This retrospective study examines the mortality patterns of a relatively young cohort of 81,746 former and current petrochemical company employees. Standardized mortality ratios (SMR) for 1979 through 1992 are generally from about unity to well below unity for major causes and numerous specific causes of death studied by gender/race/job subgroups. Findings of note include a SMR (based on incidence rates) of 1.94 (95% confidence interval [CI], 1.04 to 3.33) for mesothelioma, and a SMR of 1.81 (95% CI, 0.90 to 3.24) for chronic lymphocytic leukemia, both among males hired before 1960. All male semiskilled operatives have a 1.6-fold increase (95% CI, 1.07 to 2.29) in motor vehicle accident deaths, with declining rates since the mid-1980s. The overall SMR for acquired immunodeficiency syndrome (AIDS) is at unity (69 deaths), with excesses in technician and office worker subgroups. Four decedents with lymphoma (code 202.8 in 9th revision ICD) had AIDS as a secondary cause of death, suggesting the need to examine secondary causes when studying lymphopoietic conditions. This routine surveillance activity provides leads regarding the presence or absence of excess mortality risk. PMID- 9343763 TI - Environmental health response clinics. A survey of program options. AB - Environmental Health Response Clinics are established in response to concerns about community exposures to hazardous situations (chemical, biological, radiological). They are developed in response to a demand for "clinical services" and operate outside the usual health care financing and delivery mechanisms. Prompted by their experience in California, the authors formed a focus group to identify possible goals and services. A mail survey of occupational-environmental health professionals was then conducted to evaluate the feasibility and priority of representative goals. The analysis suggests that services should focus on the specific hazard of concern and that communication and education are essential components. The tendency to "do a general physical examination" should be eschewed. Ratings for priority and feasibility were disparate for several possible goals. In some instances, a "hands-on examination" may not be the best use of resources. Environmental health professionals may serve by direct clinical service or by advising community-based practitioners. Providing routine clinical services alone cannot meet the expectations for an environmental health response clinic. PMID- 9343764 TI - De Quervain's tenosynovitis. Stenosing tenosynovitis of the first dorsal compartment. AB - De Quervain's tenosynovitis is a disorder characterized by pain on the radial (thumb) side of the wrist, impairment of thumb function, and thickening of the ligamentous structure covering the tendons in the first dorsal compartment of the wrist. It is precisely defined as stenosing tenosynovitis of the first dorsal compartment. It is a relatively common, uncomplicated, and noncontroversial musculoskeletal disorder of the distal upper extremity. The purpose of this review is to summarize information from the medical literature on aspects of De Quervain's tenosynovitis likely to be of interest and relevant to occupational medicine practitioners. The topics covered include normal anatomy and kinesiology; history; clinical observations related to diagnosis; pathology; pathophysiology; clinical observations on etiology; descriptive epidemiology; epidemiological studies; and case management. Models for the pathogenesis of De Quervain's tenosynovitis are proposed and opportunities for future research presented. PMID- 9343765 TI - An epidemic of lung cancer due to chloromethyl ethers. 30 years of observation. AB - A cohort of 125 chemical workers was established in 1963 to investigate an epidemic of lung cancer caused by industrial exposure to chloromethyl ethers (CME). Ninety-three of the men were exposed to CME, and approximate estimates of exposure were made. Standardized mortality ratios (SMRs) were calculated for lung cancer, based on Philadelphia city rates. Over 30 years of observation, 25 of 67 deaths were due to lung cancer, with a dose-response relationship. SMRs were elevated only among 59 moderately and heavily exposed workers, peaked at 23.1 in the first decade, and then declined to 7.4 and 7.9 in later decades. The mean latency period from onset of exposure to death was 21 to 25 years and was inversely related to cumulative exposure. Three of 12 heavily exposed cases occurred in nonsmokers. Small cell carcinoma accounted for 80% of the moderately and heavily exposed cases. PMID- 9343766 TI - Carcinoma of the tonsil: prognostic factors. AB - OBJECTIVES: This study was conducted to provide a review of the prognostic factors of tonsillar carcinoma. DESIGN: A retrospective analysis. SETTING/PATIENTS: Patients with squamous cell carcinoma of the tonsil, treated in Northern Alberta, at the Cross Cancer Institute from 1975 to 1995 were analyzed using a population-based, head and neck cancer registry. There were 102 patients, 73 male and 29 female, ranging in age from 35 years to 83 years, with a mean of 60 years. The clinical stages were T1: 5 patients; T2: 27 patients, T3: 33 patients; T4: 11 patients; and Tx: 3 patients. The nodal stages were N0: 33 patients, N1: 26 patients; N2: 34 patients, N3: 7 patients; Nx: 2 patients. METHOD: The patients were treated with various modalities: surgery alone: 2 patients; surgery plus radiation: 26 patients; radiation treatment alone: 61 patients; and others: 13 patients. Patients were classified according to the UICC TNM 1992 criteria. The overall 5-year Kaplan-Meier survival in our series was 39%. The cause-specific 4-year survival was 57%. Various prognostic factors and their impact on survival were studied. RESULTS: On univariate analysis, the following factors were found to be significant. Age < 50 vs. > 50 (p = .02); endophytic growth pattern vs. exophytic growth of the primary (p = .01); ulcerated lesions vs. nonulcerated lesions (p = .000); various T stages (p = .003); clinical extension vs. no extension of primary disease (p = .02); combined modality of treatment (surgery and radiation treatment) had the best chance of survival compared to radiation treatment alone (p = .03). Nodal stages N0 vs. N+ disease (p = .2); sex of the patient, female vs. male (p = .83); and dose of radiation treatment < 5000 cGy vs. > 5000 cGy (p = .41) were found not to be significant. When the above significant factors were stratified according to the stage of the disease, only two were significant; ulcerated lesions vs. nonulcerated lesions (p = .04), and the modality of treatment chosen (e.g., radiation alone vs. radiation plus surgery) (p = .02). CONCLUSIONS: In this series of patients, combined-modity approach using surgery and radiation treatment was found to be the best way to treat carcinoma of the tonsil. However, each treatment strategy should be individualized taking into account various prognostic factors. PMID- 9343767 TI - Prevalence, density, and manifestations of oral Candida albicans in patients with Sjogren's syndrome. AB - OBJECTIVE: Various investigators have reported a high prevalence of oral Candida species in patients with salivary gland dysfunction (SGD). The purpose of this study was to assess the prevalence of oral Candida albicans, its oral manifestations, and to compare the number of colony-forming units of Candida albicans in patients with primary Sjogren's syndrome and secondary Sjogren's syndrome with the whole unstimulated salivary flow rate in each group. METHOD: An age-sex-matched group of control subjects was selected for comparison. Oropharyngeal collection of samples and culturing was performed on each subject. Quantitative cultures specific for Candida albicans were performed. RESULTS: The frequency distribution indicated that > 80% of all SS subjects were positive for Candida albicans vs. none of the controls. The most common lesion was angular cheilitis followed by chronic atrophic candidiasis. The subjects with Sjogren's syndrome also demonstrated significantly high numbers of Candida albicans colony forming units. CONCLUSIONS: This study indicates significantly higher Candida albicans colonization in patients with primary or secondary Sjogren's syndrome as compared to a control population. Candida albicans colonization was higher in patients with secondary Sjogren's syndrome than in patients with primary Sjogren's syndrome; however, the amount of Candida albicans was not universally relative to salivary flow. PMID- 9343768 TI - Effect of upper-airway passages on craniofacial growth in an animal model: a pilot study. AB - OBJECTIVE: The process of postnatal growth and development of the face and skull is of major interest to otolaryngologists. Surgery is often considered as an option for the treatment of benign and malignant tumours, traumatic facial deformities, and congenital abnormalities of the head and neck in children and adolescents. The extent of surgery and the type of reconstruction is frequently influenced by concerns about the potential effect on future craniofacial growth. Surgery is also sometimes recommended as a method to influence facial growth as in tonsillectomy and adenoidectomy for 'adenoid facies syndrome.' There are a number of different theories concerning the factors that influence the growth of the face and cranium. None of these is universally accepted. The predominant theory is the functional matrix theory. This study was designed to evaluate the validity of this theory in an animal model. A new animal model had to be developed to perform the study. This pilot study was then conducted. METHOD: A laryngotracheal separation procedure was performed on juvenile goats. This effectively eliminated the use of the upper airway by the animals, thereby removing one of the major functional matrices from the model. The animals were allowed to grow. A control group was used, and comparisons were made between the two groups. RESULTS: The results of the study suggest that the functional matrix theory is not valid in this experimental model. CONCLUSIONS: Further research is required to confirm this finding. This would have important implications for our understanding of the biology of craniofacial growth and have clinical ramifications for otolaryngologists and other clinicians with an interest in the head and neck. PMID- 9343769 TI - The integrated light endoscope: a technical simplification of nasal endoscopy. AB - OBJECTIVE: Nasal endoscopes depend on cumbersome light generators and fibre-optic cables. This results in restriction of the operator's movements, impairment of tactile sensation, and visual field limitation during the examination. More importantly, its use is difficult outside a clinic setting. A system which integrates a readily available, portable, and inexpensive light source with a nasal endoscope was tested in our department. METHOD: Twenty patients underwent endoscopic examination of their nasal cavities using this simple endoscopic system followed by the traditional light-cable technique. RESULTS: In one patient, the visual information was insufficient with the new system. In all other cases, no additional information was demonstrated by the use of light cables. CONCLUSIONS: The advantages and disadvantages of the system are discussed, as well as the future possibilities suggested by this development. PMID- 9343770 TI - Castleman's disease of the parotid gland. PMID- 9343771 TI - The fibre-optic bronchoscope as an aid to difficult tracheostomy. PMID- 9343772 TI - Carcinoma of the colon presenting as tonsillar metastasis. PMID- 9343773 TI - Cerebral tissue heterotopia in the soft palate. PMID- 9343774 TI - Skull base aspergilloma. PMID- 9343775 TI - Small-cell anaplastic carcinoma of the parotid gland. PMID- 9343776 TI - Epidemiology and etiology of hearing impairment among infants and children in a developing country. Part I. AB - OBJECTIVES: The study identifies children at risk for hearing impairment and determines the etiology, type, degree, and onset of deafness of Saudi children living in the city of Riyadh. The relationship to other anomalies is also explored, and a review of the literature is included. METHOD: This study features an extensive screening programme involving interviews, clinical and laboratory examinations, anthropometric and audiologic measurements, and family demographic data gathering of subjects both in the field and in clinics randomly selected throughout Riyadh. A control group of normal-hearing children was also selected. RESULTS: Of the 6421 children surveyed (55% male), aged 2 months to 12 years, the mean birth weight was 3050 g; the average number of siblings was 5.39; and the majority were from consanguineous families. Male children, lower birth weights, ocular problems, consanguinity, perinatal problems, lower socioeconomic level, and a family history of hearing impairment or other disease (e.g., meningitis) correlated with an increased risk of impairment in these children, as did the attendance of their mothers at an antenatal clinic. CONCLUSION: The study points to the significance of hearing impairment and its effect on communication and psychological and educational development, as well as the necessity for programmes to address these issues in children, both in management and prevention. PMID- 9343777 TI - The history of otolaryngology in the Canadian military: from first to last. PMID- 9343778 TI - The history of otolaryngology in Canada: Dalhousie University. PMID- 9343779 TI - Outcome of Perthes' disease in unselected patients after femoral varus osteotomy and splintage. AB - We evaluated 56 hips (48 patients) with Perthes' disease to compare the radiographic results of two unselected groups: one treated with femoral varus osteotomy (22 hips) and another with Thomas splint (34 hips). The patients with less than 50% femoral head involvement (Salter Group A hips) seemed to have no advantage from the operation. The angle of the femoral neck was 10 degrees less in the operative group than in the nonoperative group. In hips with more than 50% head involvement (Salter Group B), the operative method resulted in slightly better coverage and sphericity of the femoral head than the conservative method. On average, the acetabular direction was similar in both groups. The authors conclude that femoral varus osteotomy may lead to residual coxa vara and does not necessarily improve the radiographic results in limited epiphyseal involvement. Neither does the operation have an effect on the acetabular direction in severe Perthes' disease. PMID- 9343780 TI - Imaging evaluation of subluxation in Legg-Calve-Perthes disease: magnetic resonance imaging compared with the plain radiograph. AB - The aim of the study was to evaluate the advantages of magnetic resonance imaging (MRI) in determining subluxation in Legg-Calve-Perthes (LCPD) disease. Twenty-six patients with unilateral LCPD received 33 MRI and plain radiographs. For each patient, acetabulum head index (AHI) was measured on both hips (affected and unaffected) in a blinded fashion. Measurements were made from the cortical bone margin on the plain radiograph and from the cartilaginous surfaces on MRI. On the unaffected side AHI was 92.8% on the plain radiograph and 85% on MRI. On the affected side, AHI was 87% on the plain radiograph and 77% on MRI. These differences were statistically significant. With regard to the unaffected side, the femoral head should be considered subluxated if AHI is less than 86% on the plain radiograph and less than 77% on MRI. On the affected side, in 14 cases the femoral head was well-contained on both the plain radiograph and MRI. In 11 patients the femoral head was subluxated both on the plain radiograph and on MRI. In 8 patients the femoral head was well-contained on the plain radiograph but subluxated on MRI. This was due to thickening of the cartilaginous portion of the femoral head, which was clearly seen on MRI. MRI appeared to be more sensitive in determining the subluxation of the femoral head during the active phase of LCPD. PMID- 9343781 TI - Magnetic resonance imaging and early remodeling of the femoral head after femoral varus osteotomy in Legg-Calve-Perthes disease. AB - We studied 21 children with Legg-Calve-Perthes Disease with a prognostically poor development, including lateralization, poor containment, anterolateral flattering, and deformation of the femoral head, as evaluated on serial magnetic resonance (MR) imaging. These children were treated with proximal femoral varus derotation osteotomy. The sphericity of the cartilaginous and bony femoral epiphysis was evaluated postoperatively on serial radiography and MR imaging. There was an early postoperative continuous spherical remodeling over a follow-up period of 3.0 years (1.0-5.1; SD, 1.3). PMID- 9343782 TI - Intracapsular pressure in congenital dislocation of the hip. AB - Intracapsular hip joint pressure was measured in six infants with congenital dislocation of the hip (CDH) or acetabular dysplasia with hip joint instability diagnosed at an average of 4.3 months of age (range: 3-8 months). In the extension and neutral rotation position, the mean pressure was 8.9 mm Hg. After reduction, obtaining stability with the hip joints in the "frog-leg" position (i.e., maximum flexion around the axis of the neck of the femur), the mean pressure was 74.6 mm Hg. When obtaining stability with the hip joints in 20 degrees of flexion, abduction, and inward rotation, the mean pressure was 104 mm Hg, and in approximately 20 degrees of flexion, abduction, and forced inward rotation it was 160 mm Hg. We conclude that these rotational positions, often used to retain the joint in CDH or hip joint instability, induce intracapsular pressures that may cause occlusion of epiphyseal-physeal vessels and thus may be responsible for the avascular epiphyseal necrosis and growth disturbance seen in these patients. PMID- 9343783 TI - Advantages and disadvantages of various access routes in sonographic diagnosis of dysplasia and luxation in the infant hip. AB - The purpose of the study is to clarify the access route that is most suitable in sonography of the infant hip for diagnosis of luxation and dysplasia. The process of luxation gives rise to deformation of the hyaline cartilaginous preformed part as well as the osseous acetabulum. These deformations are located in the superoposterior part of the acetabulum. With a posterior exposure only the luxated head of the femur can be imaged sonographically, but the dysplastic acetabulum cannot be diagnosed. In anterior exposures the pathological changes in the superior part of the acetabulum are detected poorly or not at all. The lateral exposure can image all deformations of the acetabulum. The strictly standardized lateral sonographic exposure is hence recommended using a standardized plane of measurement. PMID- 9343784 TI - "Immunity" of Ethiopian Jews to developmental dysplasia of the hip: a preliminary sonographic study. AB - Developmental dysplasia of the hip (DDH) is one of the most common problems affecting the pediatric musculoskeletal system. The condition represents a broad spectrum of sonographic (anatomic) pathoanatomical situations, ranging from very mild dysplasia to full dislocation, manifesting clinically as stable or unstable hip joints, particularly in the neonatal period. The cause of DDH is unclear. Various theories suggest genetic, hormonal, mechanical, and geographic factors, along with hemotypology and a great variety of other factors, as causes for this condition. In the present study, sonographic criteria for diagnosing DDH according to Graf were used. Although the overall incidence of DDH was 5.9% in white neonates, it was only 0.44% (two hips, one stable and one unstable) among 450 hips of black Ethiopian newborns. Left untreated, these hips became clinically and sonographically normal. Our results could be considered to indicate a zero incidence of DDH among Ethiopian Jews in this series. PMID- 9343785 TI - Developmental dysplasia of the hip in Marfan syndrome. AB - Among 235 consecutive patients with Marfan syndrome, four cases of developmental dysplasia of the hip were found, amounting to an incidence of 2%. Harness treatment was not effective in any of the patients because of a narrow stable zone and knee laxity. One patient died of cardiac failure before further treatment could be attempted. Closed reduction and spica cast treatment was successful in all other patients. Stability was achieved within a normal period, and there were no redislocations. Consequently, the dislocated hip in patients with Marfan syndrome should be managed early with this method. PMID- 9343786 TI - Eosinophilic granuloma of the spine. AB - Twenty patients treated for eosinophilic granuloma of the spine were studied. Only 40% demonstrated the classical radiographic picture of vertebra plana. In 60% a lytic lesion of the vertebral body or the posterior elements was found. Seven patients underwent surgery; the indications were neurological involvement or failure of the biopsy to disclose the diagnosis. At an average follow-up period of 7 years, 17 patients are well and alive with no residual spinal pain, neurological compromise, recurrent disease, or extraskeletal involvement. Vertebral body collapse underwent some regeneration but did not regain full body height. In several patients this resulted in a local deformity. In patients with unifocal spinal eosinophilic granuloma, watchful observation with no treatment other than spinal support is warranted. In patients with neural involvement or multifocal lesions, a more active treatment, including surgery, may be indicated. PMID- 9343787 TI - Soft tissue behavior during limb lengthening: an experimental study in lambs. AB - The effect of femoral elongation on skeletal muscle, nerves, and vessels was studied. Three groups of five lambs were used. After the intervention, the animals were killed at 2, 3, and 4 months. A left femoral elongation of 6 cm was practiced on all of them by means of callotasis, with a distraction rate of 0.5 mm every 12 hours. The femoral elongation process was evaluated by monthly x-ray films. The nucleic acid and protein levels in the muscular tissue were quantified at the level of the elongation focus and in the control extremity. The motor conduction velocity of the sciatic nerve was measured in both posterior limbs before the intervention and immediately before the lambs were killed. The arterial blood flow of both subsequent extremities was measured at the moment of death. A histological study of quadriceps muscle, sciatic nerves, artery, and subsequent femoral vein were examined histologically at the level of the elongation focus of both extremities. After elongation, no significant differences were observed in the muscle protein and nucleic acid levels with respect to the control extremity. No significant changes of the nerve conduction velocity were observed in any animal among the different groups. The arterial blood flow of the elongated extremity showed a progressive increase, reaching its maximum value 1 month after the distraction had terminated, with subsequent normalization. This increase of the blood flow was also observed in the control extremity, suggesting a possible systemic effect. The histological study revealed a comparative thickening of the endomysium and perimysium in the elongated muscle tissue, present at the end of the distraction and which was later normalized. No histological changes of the nerve stems undergoing distraction were observed either. During elongation, the arteries showed minimal histological changes. On the other hand, the veins showed areas of endothelial damage accompanied by thrombosis phenomena, especially at the end of the distraction period. The vascular morphology presented progressive normalization after the distraction phase. PMID- 9343788 TI - Peripheral osteoarticular tuberculosis in children: tumor-like bone lesions. AB - Osteoarticular tuberculosis in children is rare in developed countries. We report three patients who were monitored between 3 1/2 years and 8 years. The primary focus was located in the distal epiphysis of the femur, distal epiphysis of the tibia, and the calcaneus. The lesions were initially radiolucent and ill defined, appearing afterward surrounded by a sclerosis halo; these features may disguise them as other infectious lesions or pseudotumoral processes. A poorly defined set of symptoms and a lack of specific signs may delay their diagnosis and treatment. Histopathological examination and culture identification are in the most accurate methods to reach a definitive diagnosis. Lesion scrapping and chemotherapy during sat least 9 months was effective in two patients. PMID- 9343789 TI - Patient variability and the design of clinical pathways after primary total hip replacement surgery. AB - Objective data are necessary for the design of clinical pathways of total hip replacement (THR) surgery. The functional recovery and timing for hospital discharge was studied in a consecutive series of 65 patients undergoing primary THR. The Modified Barthel Index (MBI) was serially measured after surgery to assess the recovery of functional independence. A MBI score of 90 out of a maximum of 100 is required before patients are fit for hospital discharge. The length of hospital stay varied from 5 days to 39 days. Fifty-eight percent of patients were fit for discharge by day 8, and 42% required 10 days or longer (mean = 14.2 days) in hospital. Patients in these two groups differed significantly with respect to age, the number of associated comorbidities, preoperative and early MBI scores, and muscle strength parameters. These data suggest that there is wide variability in patients presenting for primary THR. One single clinical pathway may not accommodate this patient variability, whereas two clinical pathways (one with a day 8 and another with an extended [day 10 +] time frame) may be more appropriate. PMID- 9343790 TI - Use of patient restraints in four Australian teaching hospitals. AB - To examine the patterns of use of patient restraints in Australian hospitals and the level of adherence to accepted guidelines, we undertook a point-prevalence study in four teaching hospitals in three different States. This involved ward inspections and review of case notes. Overall, 51 (12.5%) of the 408 people audited were being restrained with a variety of physical and chemical agents. The rate of restraint use varied from 8.5% to 18.5% between hospitals. Although the overall prevalence of restraint use increased with age, the hospital with the oldest patients used restraints least. At all hospitals, there was scant documentation in the case notes concerning the use of restraints. The prevalence of restraint use varies widely in different hospitals. As this is not explained by the patient profile, it probably reflects different philosophies of care. Documentation of the use of restraints needs to be improved in all the centres studied. PMID- 9343791 TI - Challenges for the quality movement. AB - Recent developments in the control of the performance of the health system are making it increasingly important that effective quality systems are in place. However, there is significant evidence that many quality programmes are not effective, and, in particular, resistance by many medical and other clinical staff continues. It is, therefore, important for people concerned with the implementation of quality programmes to look at the reasons why quality efforts are not meeting expectations. Areas that need attention include the willingness to apply failure analysis to programmes, recognition of the characteristics of the professional service environment, and the implications for the organizational and management context of effective quality programmes. PMID- 9343792 TI - Prevalence of obesity in surgical patients: a comparative survey in the United States and Australia. AB - STUDY OBJECTIVE: To determine and compare the prevalence of obesity in adult patients undergoing surgery in an Australian and a United States teaching hospital. DESIGN: Retrospective and prospective surveys. SETTING: Operating theatres at two university hospitals. INTERVENTIONS: Patients scheduled for surgery during two consecutive months at Royal Perth Hospital (RPH) in Perth, Western Australia, and Montefiore-University Hospital (MUH) in Pittsburgh, Pennsylvania, were studied. Age, sex, American Society of Anesthesiologists class, height, and weight data were collected from anaesthesia records. Body mass index (BMI) was calculated according to the formula: BMI = weight/height2 (kg m 2). Obesity was defined as BMI > or = 30, morbid obesity as BMI > or = 35, and overweight as BMI = 25-30. RESULTS: Data from 1604 patients were analysed. Patients ranged in age from 15 to 93 years (mean +/- SD = 52.4 +/- 20). The RPH group was slightly older (RPH = 54 +/- 20 years; MUH = 50 +/- 19 years). Men from MUH were significantly taller (MUH = 176 +/- 8 cm; RPH = 174 +/- 9 cm) and MUH women were also significantly taller (MUH = 162 +/- 8 cm; RPH = 160 +/- 8 cm) than Australian patients. Mean weight was significantly different between hospitals (RPH = 73.5 +/- 16 kg; MUH = 77.9 +/- 20 kg). Mean BMI was also significantly different between hospitals (RPH = 25.8 +/- 5; MUH = 27.3 +/- 7). The proportion of obese men (RPH = 15.7%; MUH = 21.0%), obese women (RPH = 21.9%; MUH = 30.2%), morbidly obese men (RPH = 2.1%; MUH = 6.8%), and morbidly obese women (RPH = 7.8%; MUH = 15.1%) was significantly greater in the MUH study population. However, the proportion of overweight patients was similar between countries. A greater proportion of women were found to be obese or morbidly obese in both hospitals. CONCLUSION: Obesity is more prevalent among surgical patients at MUH than at RPH, and is more common among women. The proportion of obesity seen in this survey is greater than results from general population surveys. PMID- 9343793 TI - Chris McCaffrey: a pioneer of quality in health care. PMID- 9343794 TI - Cost-effectiveness of repeat medical procedures: kidney transplantation as an example. AB - The constraints on medical-care resources can give rise to the question of the cost-effectiveness of permitting repeat medical procedures when some patients may die without undergoing even a first procedure. Using kidney transplantation as an example, this study estimates the cost-effectiveness of patients' having available the option of a repeat medical procedure in the event the first procedure fails. Specifically, the analysis examines the effect on transplant candidates of having the option of kidney retransplantation, if and when retransplantation might be needed. Data sources include the U.S. Renal Data System (USRDS) Case-Mix Severity Study, Health Care Financing Administration (HCFA) data, and a MEDLINE search. Outcome measures include life expectancy, quality-adjusted life expectancy, lifetime costs of medical care, and marginal cost-effectiveness from a societal perspective. By avoiding lifelong dialysis after graft failure, first-transplant candidates gain an average of 47 quality adjusted days with a retransplantation policy, despite the prolongation of time to first transplant by an average of 30 quality-adjusted days. The lifetime cost of medical care per first-transplant candidate is $1,210 higher with a retransplantation policy compared with the no-retransplantation policy; its societal cost-effectiveness is estimated to be $9,656 per quality-adjusted life year saved. The retransplantation policy provides the greatest improvement in quality-adjusted life expectancy for younger candidates. In the case of kidney transplantation, the cost-effectiveness of a repeat transplant, on average, compares favorably with those of other medical strategies in common practice. As resources become increasingly constrained, this study demonstrates a framework for considering the cost-effectiveness of repeat medical procedures. PMID- 9343796 TI - Incorporating future costs in medical cost-effectiveness analysis: implications for the cost-effectiveness of the treatment of hypertension. AB - It has been shown that the difference between consumption and production during life years gained should be included as a cost in cost-effectiveness analysis. In this study the authors estimate the impact of including these future costs on the cost-effectiveness of the treatment of hypertension in Sweden. The cost per quality-adjusted life year (QALY) gained changes little among young men and women due to the addition of future costs, but increases by about $14,000 for middle aged men and women and about $27,000 for older men and women. When future costs are not included, the cost per QALY gained is generally lowest among older men and women, but when future costs are included, the cost per QALY gained is generally lowest among middle-aged men and women. The authors conclude that the total resource consequences of changes in mortality should be routinely considered in cost-effectiveness analyses. PMID- 9343795 TI - The cost-effectiveness of fluconazole prophylaxis against primary systemic fungal infections in AIDS patients. AB - OBJECTIVE: To project the cost-effectiveness of fluconazole for prophylaxis against AIDS-related primary systemic fungal infections. DESIGN: A Markov model with data from the literature. PATIENTS: Hypothetical cohort of 100,000 AIDS patients. INTERVENTION: No prophylaxis, and fluconazole prophylaxis beginning when a patient's CD4 count declined to below 200/mm3, below 100/mm3, or below 50/mm3. RESULTS: The no-prophylaxis policy was associated with a discounted life expectancy of 28.20 months and direct medical costs of $36,100 per person. The < 200/mm3 strategy increased costs to $40,500 and life expectancy to 28.42 months, producing a ratio of $240,000 per year of life saved (YLS). Compared with the no prophylaxis and < 200/mm3 policies, the intermediate alternatives were less economically efficient. A reduction in fluconazole's cost from $206 to $80 decreased the ratio to $50,000 for the < 200/mm3 strategy. Doubling fungal infection incidence lowered this ratio to $96,000/YLS. CONCLUSIONS: Fluconazole prophylaxis is unlikely to be cost-effective unless its cost is lowered, or it is focused on patients in regions endemic for fungal infections. PMID- 9343798 TI - Are methods for estimating QALYs in cost-effectiveness analyses improving? AB - OBJECTIVES: The objectives of this study were to examine variations in the methods used by researchers to estimate QALYs in published cost-effectiveness analyses, and to investigate whether the methods have improved over time. DATA AND METHODS: Using a MEDLINE search, the authors identified 86 original cost effectiveness analyses, published between 1975 and 1995, that used QALYs as the measure of effectiveness. For each study, they recorded the health-state classification system, the source of the preference weights, the measurement technique, and the discount rate. The methods used were compared with the recommendations of the U.S. Panel on Cost-Effectiveness in Health and Medicine. RESULTS: Only 20% of the studies used "generic" health-state classification systems (e.g., health utilities index); 21% relied on community-based weights; 40% used formal measurement techniques (e.g., time-tradeoff method); and 88% discounted both future costs and QALYs. There was little evidence that methods had improved over time. CONCLUSIONS: The results illustrate extensive variation in the construction of QALYs in cost-effectiveness analyses and reveal that most studies have not adhered to practices now recommended by leaders in the field. There is a need for more methodologic rigor and consistency if the results of such studies are to be compared and used for purposes of allocating resources. PMID- 9343797 TI - Assessing uncertainty in cost-effectiveness analyses: application to a complex decision model. AB - A framework for quantifying uncertainty about costs, effectiveness measures, and marginal cost-effectiveness ratios in complex decision models is presented. This type of application requires special techniques because of the multiple sources of information and the model-based combination of data. The authors discuss two alternative approaches, one based on Bayesian inference and the other on resampling. While computationally intensive, these are flexible in handling complex distributional assumptions and a variety of outcome measures of interest. These concepts are illustrated using a simplified model. Then the extension to a complex decision model using the stroke-prevention policy model is described. PMID- 9343799 TI - A normative analytic framework for development of practice guidelines for specific clinical populations. AB - BACKGROUND: A central problem in practice guideline development is how to develop guidelines that appropriately account for variations in clinical populations and practice settings. Despite recognition of this problem, there is no formal mechanism for assessing what the need is for flexibility in guidelines, or for deciding how to incorporate such flexibility into recommendations. OBJECTIVE: This research sought to provide a formal basis to determine when clinical circumstances vary sufficiently that guideline recommendations should differ, how recommendations should be tailored for a specific clinical setting, and whether the benefit associated with such site-specific guidelines justifies the expense of their development. RESULTS: The authors describe an approach for estimating the maximum health benefit that developers can obtain by eliminating uncertainty about differences in the patient populations and practice settings in which a guideline will be used. This estimate, the expected value of customization, provides a mechanism to evaluate the cost-effectiveness of the development of site-specific guidelines that account explicitly for variation in clinical circumstances. Application of this method to the development of screening guidelines for human immunodeficiency virus (HIV) infection indicates that the development of site-specific guidelines potentially is cost-effective. Site specific guidelines either improve, or leave unchanged, the efficiency of HIV screening; whether they increase or decrease total expenditures and health benefits depends on the choice of a cost-effectiveness threshold, and the clinical problem. CONCLUSIONS: Development of guideline recommendations based on decision models provides a normative approach for evaluating the need for and the cost-effectiveness of site-specific guidelines that have been tailored to specific practice settings. Such site-specific guidelines can improve substantially the expected health benefit and the economic efficiency of practice guidelines. PMID- 9343800 TI - A survey of clinicians' opinions regarding the value of published decision analyses as sources of clinically useful information. AB - Published decision analyses avoid many of the practical problems thought to be contributing to the slow acceptance of clinical decision analysis. To assess clinicians' opinions regarding the usefulness of published decision analyses, 46 physicians at a large community teaching hospital judged how useful 13 proposed interventions would be in helping them make better clinical decisions. Although 48% of the respondents indicated that they clearly understood decision analysis, easy access to a published decision analysis was the lowest-ranked intervention, with 28% of the respondents indicating that it would be helpful. In contrast, 87% indicated that easy access to the latest review article, the highest-rated intervention, would be helpful. This finding suggests that the proposed practical barriers to the acceptance of clinical decision analysis are relatively unimportant. The success of efforts to foster clinical decision analysis will depend on the identification of the key factors impeding its acceptance by clinicians and the development of effective techniques to overcome them. PMID- 9343801 TI - Optimizing sampling strategies for estimating quality-adjusted life years. AB - Accurate estimation of quality of life is critical to cost-effectiveness analysis. Nevertheless, development of sampling algorithms to maximize the accuracy and efficiency of estimated quality of life has received little consideration to date. This paper presents a method to optimize sampling strategies for estimating quality-adjusted life years. In particular, the authors address the questions of when to sample and how many observations to sample at each sampling time, assuming realistically that the sample variance of quality of life is not constant over time. The method is particularly useful for the design problems researchers face when time or research budget constraints limit the number of individuals that can be surveyed to estimate quality of life. The article focuses on cross-sectional sampling. The method proposed requires some knowledge of survival in the population of interest, the approximate variances in utilities at various points along the curve, and the general shape of the quality adjusted survival curve. Such data are frequently available from disease registries, the literature, or previous studies. PMID- 9343803 TI - Application of treatment thresholds to diagnostic-test evaluation: an alternative to the comparison of areas under receiver operating characteristic curves. AB - Diagnostic tests are often evaluated by comparison of the areas under receiver operating characteristic (ROC) curves. In this study the authors compared this approach with a more direct method that takes into account consequences of a diagnosis. Data from a prospective study of diagnosis of pulmonary embolism were used for a motivating example. Using multivariable logistic regression analysis, three diagnostic models were built and compared based on their ROC curves. Although model 1 (0.706) and model 2 (0.702) had the same ROC-curve area, they performed differently when risks and benefits of subsequent decisions were considered by applying the treatment probability threshold. Models 1 and 3 (0.611) had substantially different ROC-curve areas but performed similarly taking into account the therapeutic consequences. This demonstrates that comparison of diagnostic tests using the areas under the ROC curves may lead to erroneous conclusions about therapeutic usefulness. To correspond to daily practice, it would be more appropriate to also consider the clinical implications in evaluating diagnostic tests. This is made feasible by explicit definition and application of a treatment threshold. PMID- 9343802 TI - Construct validities of the Quality of Well-Being Scale and the MOS-HIV-34 Health Survey for HIV-infected patients. AB - This research assessed the construct validities of two health-related quality-of life instruments: the Quality of Well-Being Scale (QWB) and the Medical Outcomes Study 34-item HIV Health Survey (MOS-HIV-34). A sample of 100 adult male, HIV infected patients, across six HIV disease classifications, was used as subjects. Four convergent validity measures of health-related quality of life were used: CD4 cell counts, beta-2 microglobulin levels, disease classification, and age. All convergent validity measures were significant for the QWB. Forty percent of the convergent validity comparisons with the MOS-HIV-34 were statistically significant. Because the two measures provide different perspectives on health related quality of life, both instruments appear to be useful in measuring health related quality of life in this patient population. PMID- 9343804 TI - Whose blood is safer? The effect of the stage of the epidemic on screening for HIV. AB - BACKGROUND: With improvements in HIV antibody test (ELISA) performance, the window of time between infection and seroconversion becomes a major source of error in HIV screening. The authors examined its impact on the false-reassurance rate (FRR). METHODS: Test sensitivity was modeled as the product of two factors: the inherent sensitivity (sensitivity when antibody is present) and the probability that antibody is present in infected blood. A model of HIV and AIDS incidence was used to derive an estimate of the probability of remaining in the seronegative window (pw) among those who are infected. With plausible assumptions, this probability approaches 0.03. The FRR was then estimated as a function of the probability of remaining in the seronegative window, the prevalence of HIV, and the inherent sensitivity of the ELISA test were estimated. RESULTS: The FRRs for two blood donor groups, one with an HIV prevalence of 0.004 and a typical probability of remaining in the seronegative window (pw = 0.03) and the other with a higher prevalence of 0.017 but fewer donors in the window (pw = 0.003), are equal (140 per million donors) if the blood is negative on a single ELISA test. After two negative tests or a single test that can detect antibody more reliably, however, the FRR is much higher in the group with the higher pw (= 120 per million compared with 50 per million), because the greater numbers of donors in the window more than offsets the lower prevalence. CONCLUSION: With improvements in inherent sensitivity of ELISA by virtue of technical progress or retesting, the prevalence of HIV infection may no longer play the critical role in degrading the results of blood screening. As inherent test performance improves, tests are increasingly likely to miss infected blood because of the seronegative-window error rather than because of measurement error. Window error plays a proportionally greater role during the early stages of HIV dissemination in a population where the incidence of new HIV infection is high relative to the incidence of AIDS. These findings may explain, in part, the recent observation that cases of transfusion of contaminated blood often take place in areas where AIDS epidemics have started recently. They also suggest that the traditional strategy of soliciting blood donors from low-prevalence populations may not always be optimal, unless such populations are truly low-risk. PMID- 9343805 TI - Patient preferences for thrombolytic therapy in acute myocardial infarction. AB - BACKGROUND: Despite extensive professional debate regarding the optimal thrombolytic therapy strategy in acute myocardial infarction (AMI), patient preferences have not been explored. METHODS: Preferences among patients with known or suspected coronary artery disease for treatment with tissue plasminogen activator (tPA) or streptokinase (SK) for AMI were determined using a questionnaire presenting GUSTO-1 trial and drug cost data. Preferences were based on consideration of 30-day mortality (M) alone, hemorrhagic stroke rate (SR) alone, overall preference (M + SR), drug acquisition costs, and the estimated annual costs of using a single agent to treat all AMIs. Cost-related responses were provided under payer designations of self, third-party insurance, and federal government. RESULTS: The response rate was 81% (101/125 patients). tPA was preferred by 84%, and SK by 66%, for M alone and SR alone, respectively (chi 2, p < 0.01). Overall preference (M + SR) favored tPA (78%, p < 0.01). tPA preference decreased to 43% considering drug acquisition costs under the self-pay option (p < 0.01 vs M + SR). Similar trends of lesser magnitude were also observed for the third-party and government-payer options. CONCLUSIONS: Under conditions of zero cost and consideration of mortality plus stroke-risk data, tPA were preferred overall due to its lower mortality. Introduction of drug-cost data significantly shifted the preference toward SK, particularly under the self-payer designation. Patient preferences for thrombolytic therapy in AMI indicate tradeoffs between clinical attributes and costs, and should assist in framing medical debate and decision making. PMID- 9343807 TI - Estimating CE ratios under second-order uncertainty: the mean ratio versus the ratio of means. AB - Two methods have been presented for estimating cost-effectiveness ratios under conditions of second-order (model) uncertainty: one method estimates a mean ratio of cost to effect (the "mean ratio" approach), and the other estimates a ratio of mean cost to mean effect (the "ratio of means" approach). However, the question of which estimate is theoretically correct has not been formally addressed. The authors show that the "ratio of means" approach follows directly from the theoretical foundations of cost-effectiveness analysis, has attractive internal consistency properties, and is consistent with a simple vector algebra approach to the problem. In contrast, the "mean ratio" approach has not been shown to follow from first principles, is internally inconsistent, and can prescribe economically inefficient choices. It is concluded that the "ratio of means" procedure should be preferred unless persuasive arguments are presented to the contrary. PMID- 9343806 TI - Hepatitis B immunization in a low-incidence province of Canada: comparing alternative strategies. AB - This study provides a comparative cost-effectiveness analysis of three universal immunization programs for hepatitis B virus (HBV). Using three theoretical cohorts of infants, 10-year-olds, and 12-year-olds, a universal immunization program was compared with a prenatal screening/newborn immunization program involving testing of prepartum women and immunization of newborns of HBsAg positive mothers. A Markov long-term outcome model used Manitoba data to estimate costs and health outcomes across the lifespan. The model was based on an HBV incidence rate of 19/100,000 and a discount rate of 5% and incorporated the most recent treatment advances (interferon therapy). Cost-effectiveness was calculated as the ratio of dollars spent per year of life saved, with costs determined from the perspective of a third-party payer. The universal infant-immunization program, although not cost-saving, was associated with a low, economically attractive cost-effectiveness ratio of $15,900 (Canadian) per year of life saved, a figure substantially lower than the ratios of $97,600 and $184,800 (Canadian) associated with the universal programs for 10- and 12-year-olds, respectively. Cost-effectiveness ratios were found to be sensitive to changes in immunization costs, HBV incidence rates, and the rate at which protective antibody levels are lost over time: If these variables move in the directions suggested by current trends, the authors anticipate an increasing economic appeal of universal programs well into the future. A universal program of HBV immunization for infants appears to be economically practical in regions where HBV infection rates are low and stable. PMID- 9343808 TI - The cost-effectiveness of HIV testing: accounting for differential participation rates. AB - OBJECTIVE: To understand the impact of differences in participation rates between infected and uninfected individuals on estimates of the cost-effectiveness of HIV screening. METHODS: Costs per infection detected are modeled as function of both prevalence and serostatus-dependent testing rates. Data from national surveillance surveys, seroprevalence studies, and other sources are employed to suggest the magnitude of results. RESULTS: Differential participation produces a near-doubling in the estimated cost per infection identified. This result is sensitive to assumptions regarding the benefits of screening for seronegatives. CONCLUSIONS: Voluntary HIV screening programs may incur prohibitive costs by over recruiting people at little risk of infection. Failure to account for differential participation can result in over-optimistic cost-effectiveness estimates. However, the relevance of this result--and the significance of both prevalence and participation as cost drivers--is overwhelmed by what is assumed about the benefits conferred to uninfected people by HIV screening. PMID- 9343809 TI - The power and pitfalls of simplifying assumptions. PMID- 9343810 TI - MDM policy regarding financial support of authors. PMID- 9343811 TI - Do the expected utility axioms hide flaws? PMID- 9343812 TI - Breast cancer screening at ages 40-49. PMID- 9343813 TI - Osteopontin, a coordinator of host defense system: a cytokine or an extracellular adhesive protein? PMID- 9343814 TI - Phylogenetic positions and assignment of swine and ovine corynebacterial isolates based on the 16S rDNA sequence. AB - The nucleotide sequences of the 16S ribosomal RNA gene (rDNA) of swine and ovine corynebacterial strains were determined. The sequences of the strains that identified as Corynebacterium pseudotuberculosis by their biochemical characteristics were homologous with each other. The phylogenetic position of C. pseudotuberculosis strains was closet to C. ulcerans and next closet to C. diphtheriae. The nucleotide sequence of another swine isolate, SC8, was similar to that of a recently proposed species, C. seminale, and a non-validated species, "C. glucuronolyticum," with about 0.01 to 0.02 evolutionary distances. Analysis of the predicted secondary structure of the 16S rRNA molecule agreed with the close phylogenetic relationships between C. pseudotuberculosis and C. ulcerans and between C. seminale and strain SC8. PMID- 9343815 TI - Anti-chemotactic activity of capsular polysaccharide of Cryptococcus neoformans in vitro. AB - In this study, we demonstrated the anti-chemotactic activity of the capsular polysaccharides (CPSs) isolated from each of the heavily (H)- and weakly (W) encapsulated strains of Cryptococcus neoformans in vitro. The capacity for activation of the alternative complement pathway (ACP) of cells of the two C. neoformans strains in fresh human sera was comparable to that of zymosan (insoluble control), whereas the capacity for generation of the chemotactic factor (CF) of the cells of the two strains in fresh murine sera was markedly lower in the order H- < W-strain than that of zymosan. Conversely, the capacities for ACP activation and CF generation of the CPSs were extremely lower than those of lipopolysaccharide (LPS, soluble control). When zymosan-activated murine serum was incubated with CPS, both CPSs inhibited CF activity dose dependently. When zymosan-activated serum was incubated with heat-killed cells of each strain of C. neoformans, H and W, the CF activity of the treated sera decreased significantly, suggesting that CPS per se did not affect the neutrophils directly, but CPS absorbed CF. On the other hand, both CPSs were shown to possess the O-acetyl groups in their molecules by 1H-nuclear magnetic resonance spectroscopy. The de-O acetylation of both CPSs increased the capacity for ACP activation to a level similar to that of LPS, and the de-O-acetylated CPS of both strains exhibited a lower ability to inhibit CF than did native CPS. Collectively, these results suggest that the anti-chemotactic activity of CPS accounts for its ability to absorb the CF which was mostly generated at the sites around the cell wall of whole cells via the ACP, thus suppressing the inflammatory response by preventing dispersal of CF to the extracellular space; and also that the O-acetyl group is partly, if any, involved in the mechanism for incompetence in ACP activation as well as the inhibition of CF. PMID- 9343816 TI - Intestinal distribution and intraluminal localization of orally administered Clostridium butyricum in rats. AB - Clostridium butyricum has been used as a probiotic in animals and humans for years, however, its fate in the intestine has not been clarified yet. We investigated the intestinal fate of C. butyricum using a selective medium and a monoclonal antibody after orally administering C. butyricum spores to rats. The number of C. butyricum, both viable and dead cells, in the intestinal contents were counted by enzyme-linked immunosorbent assay (ELISA) at various times after a single oral administration. The total viable number of C. butyricum was counted using a selective medium, and viable resting spores were selectively detected by treating the samples with ethanol. To investigate the intraluminal localization of the C. butyricum cells, frozen intestinal tracts were imprinted onto slides and stained with immunogold-silver. Total viable spores exceeded the number of viable resting spores by more than 10-fold from the proximal to middle of the small intestine 30 min after administration. Vegetative cells of C. butyricum were first detected in the distal small intestine after 2 hr, and vegetative growth was observed from the cecum to the colon 5 hr after administration. Dead vegetative cells were detected 9 hr after administration, and C. butyricum cells were not detected in the intestine after 3 days. The C. butyricum cells in the intestinal imprints were stained specifically by immunogold-silver staining, and proliferative cells were observed in the cecum after 3 hr. These results suggest that the administered C. butyricum germinated in the upper small intestine, grew mainly from the distal small intestine to the colon and were excreted from the rat intestine within 3 days. PMID- 9343817 TI - Deletion in the genes encoding outer surface proteins OspA and OspB of Borrelia garinii isolated from patients in Japan. AB - We detected the expression of outer surface proteins OspA and OspB, and characterized the genes encoding the two Osps of eight Borrelia garinii isolates from patients in Japan. Six of the eight strains shared a common antigenic epitope in their OspA and/or OspB proteins to monoclonal antibody P3134 against OspB, and were identified to have a conserved carboxyl terminus on their ospA and ospB genes by Southern blot hybridization. One strain, JEM4, did not express OspB protein, which was due to lack of the ospB gene. Gene cloning and sequencing analysis revealed that it had only one osp open reading frame with 819 nucleotides, which was similar to the ospA gene. The deletion of the ospB gene could be explained by a homologous recombination based on the common C-terminal sequences on the ospAB operon. PMID- 9343818 TI - A long-term survey of methicillin-resistant Staphylococcus aureus in the oral cavity of children. AB - Methicillin-resistant Staphylococcus aureus (MRSA), an indigenous bacteria in healthy people, often causes nosocomial infections. If the host human becomes compromised, MRSA can cause a serious infection. The long-term colonization of MRSA increases this risk. The purpose of this study was to demonstrate the incidence of S. aureus and MRSA colonization in the oral cavities of healthy children, and to examine the stability of identical strains of MRSA over a long term period. Fourteen children were examined in two stages (first stage: 1987-88, second stage: 1992-93). Five of the 14 children were negative for S. aureus in both stages, seven children were positive in both stages and two children were positive in only the second stage. The children who were colonized with S. aureus in the first stage always harbored the bacteria in the second stage. Of the seven children that were positive for S. aureus in both stages, three persisted in carrying MRSA. We compared two MRSA strains isolated from the same children in both stages by coagulase typing, antibiogram typing and DNA fingerprinting. In two children, the strains showed the same coagulase types, similar antibiograms and similar DNA fragment profiles. These data strongly suggest that identical strains of MRSA persisted in the oral cavities for more than five years, and that the oral cavity can serve as a reservoir for MRSA with the potential to cause nosocomial infections. PMID- 9343819 TI - Combined use of ribotyping, PFGE typing and IS431 typing in the discrimination of nosocomial strains of methicillin-resistant Staphylococcus aureus. AB - We have previously reported the phenotypic characterization of methicillin resistant Staphylococcus aureus (MRSA) clinical strains isolated in Malaya University Hospital in the period 1987 to 1989 using antibiogram, coagulase typing, plasmid profiles, and phage typing. Here, we report the analysis of the same strains with three genotyping methods; ribotyping, pulsed-field gel electrophoresis (PFGE) typing, and IS431 typing (a restriction enzyme fragment length polymorphism analysis using an IS431 probe). Ribotyping could discriminate 46 clinical MRSA strains into 5 ribotypes, PFGE typing into 22 types, and IS431 typing into 15 types. Since the differences of the three genotyping patterns from strain to strain were quite independent from one another, the combined use of the three genotyping methods could discriminate 46 strains into 39 genotypes. Thus, the powerful discriminatory ability of the combination was demonstrated. PMID- 9343820 TI - RobA-induced multiple antibiotic resistance largely depends on the activation of the AcrAB efflux. AB - RobA is a member of the XylS/AraC subfamily of DNA binding proteins, and when overexpressed, it induces multiple antibiotic resistance in Escherichia coli. In this study, we introduced a multicopy robA plasmid (pMEP1) and its derivative into OmpF mutants and an AcrAB-deficient mutant. We found that a decrease in susceptibility to multiple antibiotics in these OmpF mutants when pMEP1 was introduced did not depend on OmpF porin expression. Interestingly, a delta ompF mutant (TK007) became more sensitive when pMEP1 was introduced. Moreover, no effect of RobA on the induction of multiple antibiotic resistance in an acrA1- mutant was observed. Therefore, we conclude that the multiple antibiotic resistance induced by the overexpression of RobA largely depends on the activation of the AcrAB efflux, as well as the activation of micF. PMID- 9343821 TI - Successful development of air-dried microplates (HP-Plates) for susceptibility testing against Helicobacter pylori isolates. AB - We have successfully developed and evaluated a new susceptibility testing procedure against Helicobacter pylori strains using air-dried microplates "HP Plates" containing eight serially-diluted anti-H. pylori agents. HP-Plate wells were reconstituted by the inoculation of 100 microliters of H. pylori cell suspensions. After incubation at 37 C for 48 hr under humidified microaerophilic conditions, HP-Plates were read visually with a circular mirror. We investigated the within-day reproducibility tests of HP-Plates using the six quality control (QC) strains we proposed. Of the 20 testings, determining the minimum inhibitory concentrations (MICs) of all the QC strains fell within +/- 1 log2 dilution ranges. When 200 clinical isolates were tested with HP-Plates and compared with the results obtained with the modified broth macrodilution method of NCCLS, more than 90% of the MICs also fell within +/- 1 log2 dilution ranges. We concluded that the HP-Plate susceptibility test method is a practical and easily applicable alternative of susceptibility testing for clinical microbiology laboratories in determining the MICs of H. pylori isolates. PMID- 9343822 TI - Interferon-alpha and -gamma differentially reduce rapid immature T-cell death by contact with HIV-1 carrier cell clones in vitro. AB - The non-antigen specific rapid cytotoxic (CT) death of immature TdT+CD4+CD8+ T cells due to contact with HIV-1 carrier T-cell clones we have found recently is a novel phenomenon. The effects of interferons (IFN) on this CT reaction were studied in vitro. Treatment of the HIV-1 carrier clones, referred to as "effectors," with IFN-alpha but not IFN-gamma, or of the susceptible immature TdT+CD4+CD8+ T cells, referred to as "targets," with IFN-gamma but not IFN-alpha, for 24 hr prior to CT testing was found to reduce the CT reaction. Simultaneously, a down-regulated CD8 expression and an up-regulated antigen expression of both major histocompatibility antigen complex class I (MHC-I) and HIV-1 gp120/gp160 in the IFN-alpha treated effector (gp120+CD8+ HPB-ALL/HIV), and/or simultaneously up-regulated antigen expression of both CD8 and MHC-I in the IFN-gamma treated target (CD4+CD8+ HPB-ALL) were found to be associated with reduced CT reaction. However, altered antigen expression in the IFN-gamma treated effectors or IFN-alpha treated targets did not affect the ultimate degree of CT reaction. This study thus suggests a possible therapeutic efficacy of IFN by reducing the direct elimination of the T-cell precursors in HIV-1 infection. PMID- 9343823 TI - Triazine dyes inhibit HIV-1 entry by binding to envelope glycoproteins. AB - We have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) from the culture supernatants of CHO-Sec cells that secreted truncated 140-kDa precursor and mature 120-kDa Env glycoproteins. The concentrated culture supernatants were applied to a column coupled with cibacron blue 3GA (CB3GA) to separate albumin from the Env proteins because CB3GA, a triazine dye, has been known to have a high affinity to albumin. Unexpectedly, Env proteins as well as albumin bound to the column, and the bound Env proteins were eluted by increasing the ionic strength using KCl. Gp120 was eluted at 0.5 0.9 M of KCl, while a higher concentration (0.9-1.5 M) was necessary for the elution of gp140. The agarose gel coupled with reactive red 120 (RR120), another triazine dye with similar characteristics, also retained both Env proteins, and the bound Env proteins could be eluted in a similar manner. In addition, these agents inhibited syncytium formation caused by HTLV-IIIB and HTLV-IIIMN. Inhibition was also seen when a virus-free fusion assay between Env protein expressed in CHO cells and fluorescent labeled SupT1 cells were used. These findings indicate that triazine dyes bind to the functional regions of Env proteins of HIV-1 that play important role(s) for HIV infection. PMID- 9343824 TI - Different susceptibility of three clinically isolated strains of Cryptococcus neoformans to the fungicidal effects of reactive nitrogen and oxygen intermediates: possible relationships with virulence. AB - We investigated the susceptibility of three clinically isolated strains of Cryptococcus neoformans with different virulences to reactive nitrogen and oxygen intermediates (RNI and ROI, respectively), representing two important mediators of macrophage microbicidal activity. All mice infected with the highly virulent strain of C. neoformans, YC-11, died within 3 to 6 weeks because of rapid multiplication of the organism in the lungs and dissemination to the brain. In contrast, a weakly virulent strain, YC-13, was almost completely eradicated from the lungs and did not disseminate to the brain, leading to survival of all infected animals during the period of observation (15 weeks). The virulence of the third strain, YC-5, was intermediate between the other two strains. To examine the susceptibility of C. neoformans to the fungicidal effect of nitric oxide (NO) and superoxide anions (O2-), the organisms were exposed to these oxidants, which were chemically generated in a cell-free system. Interestingly, the number of live YC-13 yeast cells was markedly reduced after exposure to NO and O2-. In contrast, YC-11 was almost completely resistant to the killing effect of these oxidants. YC-5 showed an intermediate susceptibility. Our results demonstrate that the resistance of C. neoformans to the fungicidal effects of RNI and ROI is related to virulence, and suggest that the resistance to nitrogen- and oxygen-derived oxidants may be one of the factors to determine the outcome of infection with C. neoformans. PMID- 9343825 TI - Negative finding in cross-protective activity of Japanese Borrelia isolates against infection with three species of Lyme disease Borrelia in outbred mice. AB - Outer surface protein A (OspA) is the most promising candidate for a component vaccine against Lyme disease caused by Borrelia burgdorferi sensu lato. Active cross-protection using a whole-cell vaccine prepared from strains belonging various OspA serotypes observed in Japan and worldwide was examined. No cross protection was obtained by heterologous OspA-serotype vaccines. Since OspA is a highly variable protein expressed by Borrelia, this suggests that immunologically different OspA serotypes need to be combined for the development of an effective vaccine in Japan. PMID- 9343826 TI - Cloning and sequencing of the Vibrio parahaemolyticus fur gene. AB - A ferric uptake regulatory gene (fur) was cloned from Vibrio parahaemolyticus WP1 by a polymerase chain reaction-based technique followed by functional complementation of a fur mutation in Escherichia coli. A sequence analysis showed that, at the amino acid level, the V. parahaemolyticus Fur protein is 81% identical with the Fur protein from E. coli and over 90% identical with those of the Vibrio species. PMID- 9343827 TI - Role of curdlan sulfate in the binding of HIV-1 gp120 to CD4 molecules and the production of gp120-mediated TNF-alpha. AB - To clarify the mechanism by which curdlan sulfate (CRDS) inhibits human immunodeficiency virus (HIV)-1 infection, we examined its influence on the binding of gp120 to CD4 molecules on T cells and macrophages, as well as on the production of TNF-alpha by gp120-stimulated macrophages (which promotes HIV-1 replication). CRDS treatment of cells not only inhibited the binding of HIV-1 gp120 to CD4+ cells, but also inhibited TNF-alpha production induced by gp120. Inhibition of HIV-1 infection by CRDS may be related to these two actions. PMID- 9343828 TI - Fc receptor-mediated phagocytosis, superoxide production and calcium signaling of beta 2 integrin-deficient bovine neutrophils. AB - Fc receptor for immunoglobulin G-mediated phagocytosis, superoxide production and intracellular calcium ([Ca2+]i) signaling of complement receptor type 3 (CR3) deficient neutrophils from a heifer with leukocyte adhesion deficiency (BLAD) were compared to those of control heifers. The mean phagocytic activity of IgG coated yeasts and aggregated bovine IgG (Agg-IgG)-induced superoxide production of CR3-deficient neutrophils were 10% and 77.9%, respectively, of those of control neutrophils. The [Ca2+]i signals in CR3-deficient neutrophils stimulated with Agg-IgG or concanavalin A were different with mean peak [Ca2+]i concentrations of 78% and 41.9%, respectively, of those of control neutrophils. These findings suggest that Fc receptor-mediated neutrophil functions are closely dependent on the presence of CR3 (CD11b/CD18) on the neutrophil cell surfaces. PMID- 9343829 TI - Effects of high-fat diet and cholecystokinin receptor blockade on promotion of pancreatic ductal cell tumors in the hamster. AB - The mechanism by which high-fat diets potentiate pancreatic cancer is not known, but pancreaticotrophic hormones such as cholecystokinin (CCK) may be involved. The effect of CCK receptor blockade on carcinogenesis during the entire promotion period was investigated in Syrian Golden hamsters fed a high- or low-fat diet and treated with N-nitrosobis(2-oxopropyl)amine (3 x 10 mg/kg at weekly intervals). One-half of the hamsters fed a high-fat diet received the CCK-A receptor antagonist devazepide (25 nmol/kg/hr) for the duration of the experiment. At 39 weeks the incidence of pancreatic malignancies was significantly higher in hamsters fed the high-fat diet than in those fed the low-fat diet (p < 0.05). Tumor incidence was not changed by CCK receptor blockade. Potentiation of pancreatic cancer by a high-fat diet in hamsters does not appear to be influenced by endogenous CCK during the tumor promotion period. PMID- 9343830 TI - DHA feeding provides host protection and prevents fibrosarcoma-induced hyperlipidemia while maintaining the tumor response to araC in Fischer 344 rats. AB - Fischer 344 rats were inoculated with fibrosarcoma tumor cells and fed diets containing 5% or 10% (wt/wt) safflower oil or 10% oil containing docosahexaenoic acid (DHA). Animals were then treated with arabinosylcytosine (araC) or saline for six days. Tumor weights were highest in animals fed 10% safflower oil and treated with saline, intermediate in animals fed oil containing DHA and 5% safflower oil and treated with saline, and lowest in araC-treated animals from all diets. Plasma cholesterol and triglyceride levels correlated highly with final tumor size, regardless of diet or treatment group. Animals fed safflower oil had lower intestinal weights than those fed DHA, which histology demonstrated to be a result of differences in villus height and crypt depth. Substantial loss of bone marrow cells occurred in all dietary groups treated with araC; however, the proportion of granulocyte-macrophage precursors remaining in the DHA animals was higher than in saline-treated animals and twofold higher than in the animals fed 10% safflower oil and treated with araC. These data suggest that, even in the face of rapid tumor growth and chemotherapeutic challenge, consumption of a diet rich in DHA can slow tumor growth, prevent hyperlipidemia, enhance bone marrow cellularity, and promote intestinal growth compared with a moderate-fat n--6-rich diet. PMID- 9343831 TI - Phytoestrogen concentration determines effects on DNA synthesis in human breast cancer cells. AB - Thirteen isoflavonoids, flavonoids, and lignans, including some known phytoestrogens, were evaluated for their effects on DNA synthesis in estrogen dependent (MCF-7) and -independent (MDA-MB-231) human breast cancer cells. Treatment for 24 hours with most of the compounds at 20-80 microM sharply inhibited DNA synthesis in MDA-MB-231 cells. In MCF-7 cells, on the other hand, biphasic effects were seen. At 0.1-10 microM, coumestrol, genistein, biochanin A, apigenin, luteolin, kaempferol, and enterolactone induced DNA synthesis 150-235% and, at 20-90 microM, inhibited DNA synthesis by 50%. Treatment of MCF-7 cells for 10 days with genistein or coumestrol showed continuous stimulation of DNA synthesis at low concentrations. Time-course experiments with genistein in MCF-7 cells showed effects to be reversed by 48-hour withdrawal of genistein at most concentrations. Induction of DNA synthesis in MCF-7 cells, but not in MDA-MB-231 cells, is consistent with an estrogenic effect of these compounds. Inhibition of estrogen-dependent and -independent breast cancer cells at high concentrations suggests additional mechanisms independent of the estrogen receptor. The current focus on the role of phytoestrogens in cancer prevention must take into account the biphasic effects observed in this study, showing inhibition of DNA synthesis at high concentrations but induction at concentrations close to probable levels in humans. PMID- 9343832 TI - Content of alpha-tocopherol in blood serum of human Papillomavirus-infected women with cervical dysplasias. AB - The studies were carried out in a group of 228 female patients with normal cytological smear and 324 patients with cervical intraepithelial neoplasia. The applied method of identification, i.e., the human Papillomavirus (HPV) digene hybrid capture system, made it possible to select a control group consisting of 168 HPV-negative patients with normal Pap smear, as well as a group of 228 HPV positive patients with cervical intraepithelial neoplasia. The high-performance liquid chromatography method was employed to evaluate the level of alpha tocopherol in the blood serum of the patients who were examined. A statistically significantly lower level of alpha-tocopherol was observed in the blood serum of HPV-positive patients with cervical intraepithelial neoplasia. The risk of dysplasia was four times higher for an alpha-tocopherol level < 7.95 mumol/l. PMID- 9343833 TI - Corn oil enhances gamma-glutamyl transpeptidase-positive foci development in the presence of phenobarbital during hepatocarcinogenesis in rats. AB - We investigated the effects of type of dietary fat and phenobarbital on gamma glutamyl transpeptidase-positive foci development. Four groups of six female Sprague-Dawley rats were initiated with diethylnitrosamine (15 mg/kg) at 24 hours of age. After weaning, they were fed nutritionally complete semipurified diets containing 15% corn oil or 5% corn oil + 10% fish oil and supplemented with 5,000 ppm vitamin E with or without phenobarbital (500 ppm) for three months. Dietary fish oil significantly increased hepatic phospholipid eicosapentaenoate and docosahexaenoate concentrations and decreased arachidonate concentration compared with 15% corn oil (p < 0.05). Corn oil (15%) significantly increased hepatic prostaglandin F2 alpha concentration compared with 10% fish oil (p < 0.05). Phenobarbital significantly stimulated glutathione S-transferase activity in both dietary fat groups (p < 0.05). In the absence of phenobarbital, type of dietary fat showed no effect on hepatic gamma-glutamyl transpeptidase-positive foci development. However, in the presence of phenobarbital, 15% corn oil significantly enhanced gamma-glutamyl transpeptidase-positive foci development compared with 10% fish oil (p < 0.05). Phenobarbital showed a strong tumor promoting action in both dietary groups. In conclusion, there was an interaction between type of dietary fat and phenobarbital on gamma-glutamyl transpeptidase positive foci development during hepatocarcinogenesis in rats. PMID- 9343834 TI - Intake of selected foods and nutrients and breast cancer risk: an age- and menopause-specific analysis. AB - The relationship between selected foods and nutrients and breast cancer risk was investigated in strata of age and menopausal status using data from a case control study on breast cancer conducted between June 1991 and April 1994 in six Italian areas. Cases were 2,569 women with histologically confirmed incident breast cancer admitted to the major teaching and general hospitals of the study areas; controls were 2,588 women with no history of cancer admitted to hospitals in the same catchment area as cases for acute, nonneoplastic, nongynecological conditions unrelated to hormonal or digestive tract diseases or to long-term modifications of diet. Dietary habits were investigated using a validated food frequency questionnaire, including 78 foods or food groups. Among food groups, bread was directly and significantly related to breast cancer risk in older women and, consequently, in postmenopause, whereas the protection conferred by fish consumption was stronger in postmenopause and that exerted by raw vegetables was stronger in premenopause. Among nutrients, unsaturated fatty acids were inversely related to breast cancer risk, the association being stronger in postmenopausal and elderly women. The pattern was similar for total fats. For starch, available carbohydrates, and total proteins, no heterogeneity emerged across strata of age and menopausal status. Among micronutrients, protection diminished with increasing age for beta-carotene and calcium, whereas no heterogeneity emerged for vitamin E. Thus this age-specific analysis of the largest investigation to date on diet and breast cancer did not show any consistent pattern of breast cancer risk in relation to selected dietary factors across strata of age and menopausal status. PMID- 9343835 TI - Fibers and breast cancer risk. AB - Data from a multicenter case-control study on breast cancer conducted in Italy were used to analyze the relationship between various types of fibers and breast cancer risk. Cases were 2,569 women with histologically confirmed, incident breast cancer; controls were 2,588 women admitted to the same network of hospitals for acute, nonneoplastic, non-hormone-related diseases. Cases and controls were interviewed between 1991 and 1994 using a validated food frequency questionnaire. The data were modeled through multiple logistic regression, controlling for demographic and reproductive breast cancer risk factors. The continuous odds ratios for the difference between the upper cut point of the fourth and the first quintile of intake were 0.90 [95% confidence interval = 0.82 0.98, p (for trend) < 0.05] for cellulose and 0.94 (95% confidence interval = 0.86-1.02) for soluble fibers. The protection tended to be stronger in premenopausal women. No material association was found for noncellulose polysaccharides and lignin. This study, based on a large data set from various Italian regions, suggests that fiber intake may confer some protection against breast cancer, particularly for cellulose and also for soluble fibers, i.e., those of vegetable origin. This possible protection has been related to an influence of fibers on levels and availability of estrogens and other steroid hormones in breast carcinogenesis. PMID- 9343836 TI - Soy feeding induces phase II enzymes in rat tissues. AB - The ability of soy to induce phase II detoxification enzymes was evaluated in male Sprague-Dawley rats. Soybeans contain biologically active compounds that are known inducers of phase II enzyme activity. Rats were fed soy flour (SF) or soy protein isolate (SPI) to provide 75% of total protein as soy. Rats were given free access to food for one- and two-week periods before enzyme activity was compared with that of casein control groups (AIN-93G). Hepatic glutathione S transferase (GST) activity was significantly greater in rats fed SF for one and two weeks and in rats fed SPI for two weeks than in controls. Quinone reductase activity was significantly greater (12- to 14-fold) in the colon of rats fed SF and SPI for two weeks and in serum (1.8- to 2-fold) in the SF group at one and two weeks. Liver, kidney, and small intestine uridine 5'-diphosphate-glucuronosyl transferase activity was significantly increased in the SPI and SF groups at two weeks. A time dependence in induction of phase II enzymes was observed in several tissues. There was no significant difference in total liver glutathione in either diet group compared with controls. The data indicate that dietary soy enhances phase II enzyme activity, especially quinone reductase and uridine 5'-diphosphate glucuronosyl transferase, which could lead to protection from potentially harmful xenobiotics. PMID- 9343837 TI - Prospective study of diet and female colorectal cancer: the New York University Women's Health Study. AB - The relation between diet and female colorectal cancer was analyzed in a prospective study of 14,727 women aged 34-65 years, who were enrolled at mammographic screening clinics in New York and Florida from 1985 to 1991. They were followed through the end of 1994 (average 7.1 yrs) by a combination of direct contact through mail and telephone and record linkages with regional tumor registries, resulting in 100 incident cases of colorectal cancer. There was no overall positive or inverse association of colorectal cancer risk with intakes of total calories, total or subclasses of fat, carbohydrate, or dietary fiber, whereas there was an inverse association with total protein. Among major food groups, there was a progressive decline in risk of colorectal cancer with increasing intake of fish and shellfish (relative risk for 4th vs. 1st quartile = 0.49, 95% confidence interval = 0.27-0.89). A similar inverse association was also observed for consumption of dairy products, and this association was explained mainly by calcium, not by other nutrients, such as fat or protein. The results of the present study indicated that certain dietary components of fish or dairy products may protect against colorectal cancer, whereas the relations with red meat or total fat remained unclear. PMID- 9343838 TI - Feasibility of a randomized trial of a high-vegetable diet to prevent breast cancer recurrence. AB - Epidemiologic evidence supports the concept that diet influences risk for breast cancer and suggests that prognosis after the diagnosis of breast cancer may also be related to modifiable nutritional factors. The purpose of this study was to investigate the feasibility of a randomized trial of a high-vegetable, reduced fat, and increased-fiber diet intervention to reduce risk for recurrence among breast cancer survivors. This major change in dietary pattern was promoted through intensive telephone counseling. Participants were 93 women who had been diagnosed with breast cancer (stages I, II, and IIIA) within the previous four years and who had completed their initial treatment. We assessed adherence to the study diet using repeated 24-hour dietary recalls at 6 and 12 months and measurement of circulating carotenoid concentrations. Six months after randomization, the intervention group had significantly increased their mean intake of vegetables (+4.6 servings/day), fruit (+0.7 servings/day), and fiber (+6.4 g/1,000 kcal) and significantly reduced their intake of dietary fat (-9.9% of energy) compared with the control group. Circulating concentrations of carotenoids also increased in the intervention group. These changes persisted at the 12-month visit. Results of this study demonstrate that telephone counseling can be a useful approach in diet intervention and that breast cancer survivors can adopt and maintain a high-vegetable, reduced-fat dietary pattern. PMID- 9343839 TI - An ecological study of trends in cancer incidence and dietary changes in Hong Kong. AB - Cancer incidence rates from the Hong Kong Cancer Registry show significant increases in lung and colon cancers and decreases in nasopharyngeal cancer in both sexes from 1973 to 1992. Moreover, cervical cancer and male esophageal cancer have declined significantly, and changes in the trends of cancer of the following sites were of borderline significance: decreasing male laryngeal and female esophageal cancers and increasing prostate and female breast cancers. These changes have occurred along with dietary shifts in the population, from a diet predominantly of rice and small portions of meat, vegetables, and fish to one with larger portions of all foods but rice and eggs. The latter data were gathered from six government household surveys from 1963-64 to 1994-95. By combining the two data sets, correlation coefficients were calculated for per capita consumption patterns of eight foods (rice, pork, beef, poultry, saltwater fish, freshwater fish, fresh vegetables, and eggs) and cancer incidence data of the same year or 10 years later. Higher meat intakes were significantly and positively correlated with cancers of the colon, rectum, prostate, and female breast. The correlations also suggested that current diets were more influential than diets a decade before for cancers of the lung, esophagus, rectum, and prostate. Cancers of the nasopharynx and colon were significantly correlated with current and past diets. These results support the hypothesis that intakes of meat and its associated fat are risk factors for colon, rectal, prostate, and female breast cancers. PMID- 9343840 TI - Anthropometric risk factors for prostate cancer. AB - Cancer of the prostate is the leading cancer among American men, yet few risk factors are known. Anthropometry may help uncover potential risk factors for prostate cancer, since fat distribution, skeletal structure, and musculature may differ between men with this hormonally linked cancer and those without it. A case-control study was undertaken to determine whether anthropometric differences exist between prostate cancer cases and controls and whether such differences are associated with specific hormonal profiles. The study accrued 315 men stratified for race, age, and case/control status. Weight, height (sitting/standing), skinfold thicknesses (triceps, biceps, subscapular, suprailiac, thigh), circumferences (midarm, waist, hip, thigh), breadths (elbow, biacromial, biiliac), hormonal levels (total and free testosterone, dihydrotestosterone, sex hormone-binding globulin), bone density, and body composition were measured. Measures of upper body robustness [i.e., biacromial breadth-to-height ratio (p = 0.02) and biacromial (p = 0.05) and bideltoid (p = 0.04) breadths] were greater among controls. Strong negative associations were found uniformly between sex hormone-binding globulin levels and measures of body adiposity and musculature. Data show that prostate cancer cases exhibit a propensity toward a slight upper body skeleton, which may in itself serve as a risk factor or provide a benchmark of past nutritional and/or hormonal status and help elucidate the etiology of this disease. PMID- 9343841 TI - Acute-phase reactants and plasma trace element concentrations in non-small cell lung cancer patients and controls. AB - This study examined the effect of an acute-phase response on plasma trace element concentrations of non-small cell lung cancer (NSCLC) patients. In normal subjects (n = 13) and NSCLC patients (n = 22), fasting concentrations of albumin, C reactive protein, the trace elements iron, zinc, copper, and selenium, and their associated proteins transferrin, albumin, ceruloplasmin, and glutathione peroxidase were measured. The NSCLC patients were subdivided into two equal groups depending on whether they had a C-reactive protein concentration < 35 mg/l (Group 1) or > 35 mg/l (Group 2). Circulating zinc, iron, and transferrin concentrations were significantly lower in NSCLC Group 1 than in the control group (p < 0.05). Circulating concentrations of iron, zinc, and the binding proteins transferrin and albumin were significantly lower in NSCLC Group 2 than in the control group and NSCLC Group 1 (zinc not significantly different) (p < 0.01). In contrast circulating concentrations of copper and its binding protein ceruloplasmin were significantly increased in NSCLC Group 2 compared with NSCLC Group 1 and the control group (p < 0.01). Additionally, plasma selenium and glutathione peroxidase concentrations were significantly lower (p < 0.05) in NSCLC Group 2 than in NSCLC Group 1 and the control group. In the NSCLC patients there were significant negative correlations between concentrations of C-reactive protein and iron, transferrin, zinc, albumin, and selenium (p < 0.05). Furthermore, there were also significant positive correlations between C-reactive protein and copper (r = 0.788, p < 0.001) and ceruloplasmin (r = 0.831, p < 0.001) concentrations. The presence of an acute-phase response has implications for the interpretation of circulating trace element concentrations, the status of patients with NSCLC, and supplementation with trace elements in patients with NSCLC. PMID- 9343842 TI - [Feasibility of "Valsalva Leak Point Pressure". Prospective study]. AB - OBJECTIVE: To prospectively evaluate the feasibility of determination of the Valsalva Leak Point Pressure (VLPP). PATIENTS AND METHODS: From 1st January to 31st July 1996, 155 consecutive patients investigated for urinary incontinence with no pelvis static disorder performed Valsalva manoeuvres during cystomanometry in order to determine the VLPP. The examination was performed in the standing position at a filling volume of 200 cc with then without a vesical pressure transducer. The mean age of the patients was 54 +/- 16 years (range: 16 84 years). RESULTS: The mean maximal intensity of abdominal straining pressure measured by the intravesical transducer was 72 +/- 28 cm of water. The VLPP could not be determined in 50.4% of cases, as the abdominal straining pressure during the Valsalva manoeuvre was less than 60 cm of water. No correlation was observed between abdominal straining pressure and patient age (r = 0.13; p > 0.1). CONCLUSION: Leak Point Pressure cannot always be determined by the Valsalva method. Other techniques of progressive increase of intravesical pressure must be investigated. PMID- 9343843 TI - Clinical aspects of pulmonary embolism. AB - Pulmonary embolism is a common disease in the United States, affecting as many as 500,000 persons annually. Unfortunately, this disorder is commonly undiagnosed, resulting in significant excess morbidity and mortality. The clinical symptoms and signs caused by pulmonary embolism are nonspecific and may be confused with a variety of other cardiopulmonary disorders having similar presentations. However, accurate diagnostic tests are available for diagnosing pulmonary embolism, even in the face of coexistent cardiopulmonary disorders. This article describes the clinical characteristics of pulmonary venous thromboembolism, reviewing its typical symptoms and signs, its routine laboratory tests, and chest radiographic abnormalities. PMID- 9343844 TI - CT of pulmonary thromboembolism. AB - Conventional incremental CT has for many years been useful in the fortuitous diagnosis of pulmonary thromboembolic disease, allowing for visualization of both the central occluding thrombus and the pleuroparenchymal sequelae. Unfortunately, the slow data acquisition times precluded the inclusion of conventional CT in diagnostic algorithms for the diagnosis of this disease. The development and increasing availability of fast scanning techniques, namely helical (spiral) CT and electron-beam CT, now provide a noninvasive means of consistently and accurately demonstrating acute and chronic pulmonary arterial thrombus to the segmental level. CT has the added advantage over ventilation-perfusion scanning and pulmonary angiography of depicting unsuspected intrathoracic disease that may account for the patient's presenting illness. PMID- 9343845 TI - Magnetic resonance imaging of pulmonary embolism. AB - Magnetic resonance imaging (MRI) has the unique ability to demonstrate pulmonary emboli, venous thrombosis, and normal pulmonary arteries in a single noninvasive study. Spin echo and gradient echo pulse sequences take advantage of the natural high contrast between flowing blood and intraluminal thrombus or embolus. Magnetic resonance angiographic (MRA) techniques offer three-dimensional display of the pulmonary vasculature. Each of these techniques may be viewed in cinematic fashion to depict hemodynamic changes associated with the cardiac cycle. Clinical studies have demonstrated sensitivity in the 75% to 100% range and specificities between 42% and 90% depending on technique. MRI technology is still rapidly advancing and clinical accuracy will no doubt improve as experience with new techniques develops. At present, MRI should play a complimentary role to conventional methods of diagnosing thromboembolic disease. PMID- 9343846 TI - MR pulmonary angiography and perfusion imaging: recent advances. AB - Recent advances in MR pulmonary angiography and MR perfusion imaging are reviewed, focusing on two principal areas of technical development: (1) the availability of MR scanners equipped with enhanced gradient systems; and (2) new trends in MR angiography using gadolinium contrast agents or labeling of blood with an inversion recovery radiofrequency pulse in place of the more traditional methods using naturally flowing spins as the source of intravascular signal. These recent developments in MR have significant potential for clinical imaging of the pulmonary vasculature, particularly for the diagnosis of pulmonary embolism, and are now opening windows to functional MR imaging of the lung. PMID- 9343848 TI - Spiral CT evaluation of deep venous thrombosis. AB - Spiral CT venography is a technical innovation in vascular imaging that can optimize vessel contrast in the deep venous system and, therefore, is an accurate diagnostic tool to detect deep venous thrombosis. Compared with conventional venography, the amount of contrast material can be reduced by 80%. While using spiral CT as the primary imaging technique for the detection of pulmonary embolism, the cause of embolism can be evaluated within a short period of additional imaging time without further patient mobilization. This review outlines fundamental techniques in spiral CT venography and summarizes our clinical experience at Vienna University Medical Center. PMID- 9343849 TI - Evaluation of algorithms for the diagnosis of pulmonary embolism. AB - The development of new diagnostic tests for diagnosing pulmonary embolism (PE) has not yet resulted in any single algorithm being universally accepted. It is at least partially due to the perception by the clinicians ordering the tests that insufficient weight is given to the actual mortality and morbidity costs of pulmonary angiography in comparison to those associated with PE and its treatment. It also becomes more difficult to intuitively integrate all of the competing factors (e.g., sensitivity, specificity, costs, morbidity, mortality) for the different algorithms to choose the most cost-effective sequence. We evaluated the algorithm recommended by the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) investigators, designated by V, and compared it to pulmonary arteriography (A), MR angiography (M), and CT angiography (C) which directly visualize the pulmonary arteries. Using standard economic approaches to loss of life and morbidity to determine the possible costs, we compared the different algorithms for all possible prevalence values. The sensitivity and specificity rates used in making these comparisons were the average values reported in the literature. We found that the recommended algorithm, V, had the lowest cost, provided that economically reasonable morbidity and mortality costs were used. It seems that this algorithm provides sufficient sensitivity and specificity to compensate for the risks of angiography as compared with the other algorithms that avoid these risks. Neither M nor C can compete with this standard algorithm despite their lower mortality and morbidity costs, and the costs of A alone are too high. In the future, however, the inclusion of venous studies with M and/or C may improve the results sufficiently to become more cost-effective than V. PMID- 9343847 TI - Controversies in the use of lower extremity sonography in the diagnosis of acute deep vein thrombosis and a proposal for a unified approach. AB - During the past 10 years, lower extremity venous ultrasonography (LEUS) has essentially replaced contrast venography for the evaluation of patients suspected of acute thromboembolic disease. Along with this change in technology, the number of studies performed to rule out DVT has increased dramatically, and there has been controversy in the literature regarding the most appropriate role of LEUS. Specifically, four questions have been raised: (1) What is the appropriate role of LEUS in patients with a nondiagnostic V/Q scan?; (2) Is there a need to examine the contralateral (asymptomatic) leg of patients presenting with unilateral DVT symptomatology?; (3) What role, if any, does LEUS play in the evaluation of patients with bilateral leg symptoms?; and (4) When can a limited examination of the leg be performed? The purpose of this article is to provide a balanced review of these issues and to present an algorithm for a reasonable, integrated approach to the use of LEUS among patients suspected of having acute thromboembolic disease. PMID- 9343850 TI - Chronic thromboembolic pulmonary hypertension: the disease, the diagnosis, and the treatment. AB - Chronic thromboembolic pulmonary hypertension is a disease of unknown etiology, the diagnosis and treatment of which has changed dramatically in the past decade. Increased clinical awareness and recent developments in imaging techniques combine to promote earlier and less invasive diagnosis. Improved surgical thromboendarterectomy techniques and decreased perioperative mortality have enabled remarkable cures for most patients with this previously fatal condition. This article reviews current understanding of the disease process, imaging modalities used in diagnosis, and surgical treatment of patients with chronic thromboembolism. PMID- 9343851 TI - The first peptide-gated ion channel. AB - Patch-clamp experiments on the C2 neurone of Helix aspersa have shown that the neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) directly gates a Na+ channel. The channel is amiloride-sensitive. Activation of this channel is responsible for the fast excitatory action of the peptide. Using primers based on amiloride-sensitive epithelial Na+ channels, a complete cDNA sequence (FaNaCh) was cloned and sequenced from a Helix library. The sequence is predicted to have just two membrane-spanning regions and a large extracellular loop. When expressed in Xenopus laevis oocytes, the channel responded to FMRFamide. Taken together, these data provide the first evidence for a peptide-gated ion channel. Comparison of the properties of the expressed FaNaCh with the native neuronal channel show small differences in the sensitivities to some drugs and in channel conductance. It is not yet clear whether the native channel is a homo-oligomer or comprises other subunits. The peptide FKRFamide is an effective antagonist of FMRFamide on the expressed and neuronal channels. Nucleotide sequences encoding similar channel proteins occur in neurones of species as dissimilar as man and Caenorhabditis elegans. Some channels are thought to be associated with mechano sensation, at least one is a proton-gated channel and others may also be ligand gated channels. PMID- 9343852 TI - Actin-based vesicular transport in the first 20 min after dusk is crucial for daily rhabdom synthesis in the compound eye of the grapsid crab Hemigrapsus sanguineus. AB - In the crab Hemigrapsus sanguineus, maintained under a 12 h:12 h light:dark cycle, the amount of vesicular smooth endoplasmic reticulum (vesicular sER) in the photoreceptor cell body increases after the light is turned off. This paper demonstrates that actin filaments in the photoreceptor cell body are involved in the transport of vesicular sER towards the rhabdom. To specify the time of actin contribution to rhabdom synthesis, we disrupted the organization of actin filaments in the cell body with cytochalasin D at various time around dusk. We then measured the rhabdom size and also examined the ultrastructure of the photoreceptor cell body 3 h after extinguishing the light. When cytochalasin D was applied from either 1 h before or immediately after extinguishing the light, the rhabdom size did not increase, whereas vesicular sER accumulated in the cell body. In contrast, cytochalasin D applied to the eyes from 20 min after turning the light off did not inhibit rhabdom synthesis. These results indicate that the first 20 min after the light is turned off is particularly important for the transport of vesicular sER towards the rhabdom by the cell body actin filaments. PMID- 9343853 TI - Transfer of the heart pacemaker during juvenile development in the isopod crustacean Ligia exotica. AB - Developmental changes in heartbeat pacemaker mechanisms were examined electrophysiologically in the isopod crustacean Ligia exotica. The heartbeat of embryos and early juveniles was myogenic. The heart muscle cells were coupled electrically, and no localized pacemaker activity was found in the heart. In newly hatched juveniles, the cardiac ganglion exhibited no spontaneous activity, although stimulation of the cardiac ganglion produced excitatory junctional potentials (EJPs) in the heart muscle. The myogenic activity of the heart was reset and entrained by the EJPs evoked by ganglionic stimulation. During juvenile development, spontaneous EJPs appeared irregularly in the heart muscle. Later in development, the cardiac ganglion started rhythmic bursting, and each muscle response followed a ganglionic burst discharge and overlapped the EJPs evoked by ganglionic activity. At this point, the activity of the cardiac ganglion was suppressed by application of tetrodotoxin (TTX); however, even in old adults, both muscle activity and the heartbeat continued following TTX application. Heartbeat frequency was lower in TTX-containing saline than in normal saline. These results show that, during juvenile development, the heart pacemaker is transferred from the heart muscle to the cardiac ganglion, which becomes the primary pacemaker and entrains the heart muscle activity to a higher frequency via EJPs. PMID- 9343854 TI - A stereological comparison of villous and microvillous surfaces in small intestines of frugivorous and entomophagous bats: species, inter-individual and craniocaudal differences. AB - The extents of functional surfaces (villi, microvilli) have been estimated at different longitudinal sites, and in the entire small intestine, for three species of bats belonging to two feeding groups: insect- and fruit-eaters. In all species, surface areas and other structural quantities tended to be greatest at more cranial sites and to decline caudally. The entomophagous bat (Miniopterus inflatus) had a mean body mass (coefficient of variation) of 8.9 g (5%) and a mean intestinal length of 20 cm (6%). The surface area of the basic intestinal tube (primary mucosa) was 9.1 cm2 (10%) but this was amplified to 48 cm2 (13%) by villi and to 0.13 m2 (20%) by microvilli. The total number of microvilli per intestine was 4 x 10(11) (20%). The average microvillus had a diameter of 8 nm (10%), a length of 1.1 microns (22%) and a membrane surface area of 0.32 micron 2 (31%). In two species of fruit bats (Epomophorus wahlbergi and Lisonycteris angolensis), body masses were greater and intestines longer, the values being 76.0 g (18%) and 76.9 g (4%), and 73 cm (16%) and 72 cm (7%), respectively. Surface areas were also greater, amounting to 76 cm2 (26%) and 45 cm2 (8%) for the primary mucosa, 547 cm2 (29%) and 314 cm2 (16%) for villi and 2.7 m2 (23%) and 1.5 m2 (18%) for microvilli. An increase in the number of microvilli, 33 x 10(11) (19%) and 15 x 10(11) (24%) per intestine, contributed to the more extensive surface area but there were concomitant changes in the dimensions of microvilli. Mean diameters were 94 nm (8%) and 111 nm (4%), and mean lengths were 2.8 microns (12%) and 2.9 microns (10%), respectively. Thus, an increase in the surface area of the average microvillus to 0.83 micron 2 (12%) and 1.02 microns 2 (11%) also contributed to the greater total surface area of microvilli. The lifestyle-related differences in total microvillous surface areas persisted when structural quantities were normalised for the differences in body masses. The values for total microvillous surface area were 148 cm2g-1 (20%) in the entomophagous bat, 355 cm2g-1 (20%) in E. wahlbergi and 192 cm2g-1 (17%) in L. angolensis. This was true despite the fact that the insecteater possessed a greater length of intestine per unit of body mass: 22 mm g-1 (8%) versus 9-10 mm g-1 (9-10%) for the fruit-eaters. PMID- 9343856 TI - The effects of seasonal hypertrophy and atrophy on fiber morphology, metabolic substrate concentration and sound characteristics of the weakfish sonic muscle. AB - Male weakfish Cynoscion regalis possess highly specialized, bilateral, striated sonic muscles used in sound production associated with courtship. Androgen-driven hypertrophy of the muscles during the late spring spawning period results in a tripling of sonic muscle mass followed by post-spawning atrophy. This study examined the morphological and biochemical changes underlying seasonal changes in sonic muscle mass and the functional effects of these on contraction as measured by sound production. Sonic muscle fiber cross-sectional area (CSA) increased significantly during the period of hypertrophy and then decreased by nearly 60%. Both the CSA of the contractile cylinder and that of the peripheral sarcoplasm decreased significantly by late summer, with the peripheral ring of sarcoplasm virtually disappearing. Muscle protein content followed a similar trend, suggesting a major loss of structural elements during atrophy. Muscle glycogen and lipid content decreased precipitously in early June during the period of maximal sound production. Sound pressure level increased and sound pulse duration decreased with increasing sonic muscle mass, indicating that sonic muscle fibers contract with greater force and shorter duration during the spawning season. Neither the pulse repetition rate nor the number of pulses varied seasonally or with muscle mass, suggesting that the effects of steroids on the acoustic variables are more pronounced peripherally than in the central nervous system. Seasonal sonic muscle hypertrophy, therefore, functions as a secondary sexual characteristic that maximizes vocalization amplitude during the spawning period. PMID- 9343857 TI - Three opsin-encoding cDNAS from the compound eye of Manduca sexta. AB - Three distinct opsin-encoding cDNAs, designated MANOP1, MANOP2 and MANOP3, were isolated from the retina of the sphingid moth Manduca sexta. MANOP1 codes for a protein with 377 amino acid residues. It is similar in sequence to members of a phylogenetic group of long-wavelength-sensitive arthropod photopigments, most closely resembling the opsins of ants, a praying mantis, a locust and the honeybee. MANOP2 and MANOP3 opsins have 377 and 384 residues respectively. They belong to a related group of insect visual pigments that include the ultraviolet sensitive rhodopsins of flies as well as other insect rhodopsins that are also thought to absorb at short wavelengths. The retina of Manduca sexta contains three rhodopsins, P520, P450 and P357, with absorbance peaks, respectively, at green, blue and ultraviolet wavelengths. There is evidence that MANOP1 encodes the opsin of P520. We suggest that MANOP2 encodes P357 and that MANOP3, representing a class of blue-sensitive insect photopigments, encodes P450. PMID- 9343858 TI - Changing patterns of vasculature in the developing amphibian retina. AB - Patterns of vascularisation were examined in whole-mounted retinae from tadpole stages to adulthood in the tree frog Litoria moorei using perfusion with Indian ink. Changing cell densities in the underlying ganglion cell layer were studied in a parallel Cresyl-stained series. Throughout development, the vasculature was pan-retinal and the hyaloid vessel was prominent. In early tadpole stages, capillaries were arranged as a honeycomb, and their number increased at a rate sufficient to maintain high densities in the face of increasing retinal area; major arteries and veins condensed within the capillary network. By early post metamorphic life, the retinal vasculature was remodelled by the loss of four fifths of the capillaries; the reduction in their density was far greater than could be accounted for by continuing retinal growth. This loss resulted in a change from the honeycomb appearance to one with largely parallel vessels linked by fewer connecting ones, an arrangement that became increasingly pronounced. In post-metamorphic life, the number of branch points increased such that their density decreased only slightly in the face of considerable increases in retinal area. The density of branch points varied across the retina and changed with age. Initially, the vasculature was most dense centrally, but by mid-larval life densities were highest in two patches located in the mid-temporal and mid-nasal retina. Thereafter, the vasculature increasingly assumed gradients resembling an area centralis and visual streak, a profile that survived the vascular remodelling. The development of density gradients in the vasculature preceded that of cells in the ganglion cell layer, the latter appearing only following metamorphosis. However, in post-metamorphic life, the topographies of the retinal vasculature and cells in the ganglion cell layer were closely related. PMID- 9343859 TI - Maltose phosphorylase from Lactobacillus brevis: purification, characterization, and application in a biosensor for ortho-phosphate. AB - With the goal to obtain maltose phosphorylase as a tool to determine ortho phosphate, the enzyme from Lactobacillus brevis was purified to 98% by an expeditious FPLC-aided procedure which included anion exchange chromatography, gel filtration, and hydroxyapatite chromatography. The native maltose phosphorylase had a molecular mass of 196 kDa and consisted of two 88 kDa subunits. In isoelectric focusing two isoforms with pI values of 4.2 and 4.6 were observed. Maximum enzyme activity was obtained at 36 degrees C and pH 6.5 and was independent of pyridoxal 5'-phosphate. The apparent K(m) values with maltose and phosphate as substrates were 0.9 mmol l-1 and 1.8 mmol l-1, respectively. Maltose phosphorylase could be stored in 10 mM phosphate buffer pH 6.5 at 4 degrees C with a loss of activity of only 7% up to 6 months. The stability of the enzyme at high temperatures was enhanced significantly using additives like phosphate, citrate, and imidazole. The purified maltose phosphorylase was used as key enzyme in a phosphate sensor consisting of maltose phosphorylase and glucose oxidase. A detection limit of 0.1 microM phosphate was observed and the sensor response was linear in the range between 0.5 and 10 microM. PMID- 9343860 TI - Significance of health fitness appraisal in an aging society. AB - There is no doubt that the older population in Japan is rapidly increasing. The over-65 age group is the fastest growing age group in Japanese society. The quality of life for this rapidly growing segment of the population can no longer be ignored without disastrous consequences. The advent of an increased life expectancy has focused attention on the issue of functionality versus disability. We are all faced with the inevitable consequences of aging, yet each of us has the capacity to modify the aging process physiologically through appropriate physical activity and other preventive health measures. Therefore, with the aid of a physically healthy lifestyle, an exercise participant can be physically capable, energetic, and live actively beyond the ages of 50, 60, or even 70 years. Consequently, a key issue for successful or healthy aging would appear to be the improvement in perception of physical ability through education, as well as improvement in health-related physical fitness through a change in lifestyle involving regular exercise. In addition, it is a major responsibility of the physical education profession and related health fields to clarify and publicize the benefits, risks, and specific parameters of physical activity, and to develop an effective prescription for physical activity in programs that are age adjusted. This review discusses from this perspective the significance of health fitness appraisal in the aged society. Much more research is needed to clarify these issues in Japanese society. PMID- 9343861 TI - Fitness, diet and coronary risk factors in a sample of southeastern U.S. children. AB - The purpose of this study was to evaluate the relationship between physical fitness variables and nutrient intake to coronary risk factors (CRF) in a sample of children living in the Southeastern U.S. A total of 22 sixth-grade children of whom 10 were boys (mean age = 11.83 +/- 0.3) and 12 were girls (mean age 11.7 +/- 0.3) volunteered for this study. Results indicated that boys in comparison to girls weighed more (54.0 +/- 10.8 kg versus 42.1 +/- 8.0 kg; p < 0.05), had a higher body mass index (BMI) (23.6 +/- 2.7 versus 20.2 +/- 3.3; p < 0.05), a higher lean body mass (37.8 +/- 6.0 kg versus 30.7 +/- 3.8 kg; p < 0.01), and a higher systolic blood pressure (115.7 +/- 11.1 versus 106.4 +/- 8.1; p < .0001). There were, however, no significant gender differences in serum lipoproteins or nutrient intake. Stepwise multiple regression analyses indicated that physical fitness variables which included VO2max, one-mile run for time, grip strength, and leg strength could significantly predict resting diastolic blood pressure (DBP) (F = 3.06; p < 0.05) and percent body fat (F = 4.98; p < 0.01) in children. Analysis of food intake revealed that total and saturated fat, and carbohydrate intake could predict serum triglycerides (TG) (F = 5.18; p = 0.01) while total kilocalorie, fat, and carbohydrate intake could significantly predict percent body fat (F = 3.42; p < 0.03). These findings may be clinically relevant since both serum triglyceride levels and percent body fat were well above the 50th percentile according to U.S. norms. In summary, the present study showed that measurements of muscular strength in addition to aerobic fitness are associated with DBP and percent body fat in children. Furthermore, it is recommended that nutrient intake be used when evaluating CRF in children due to its ability to predict TG and percent body fat. PMID- 9343862 TI - Bone changes due to hyperbaric exposure. AB - Based on the hypothesis that bone calcification is promoted by loading physical pressure, changes in the microstructure of the bone under hyperbaric conditions were analyzed by imaging technology. Hyperbaric exposure was carried out for two weeks at 2 atm (equal to the pressure at a depth of water of 10 m) which was achieved using a mixed gas of helium and oxygen (He:O2 88%:12%) in which the oxygen partial pressure was maintained at constant (PO2: 0.21 bar). In image technological analysis, the growth and development of the bone were evaluated at different stages using Digital Magnification Radiography (DMR) images and based on changes in the X-ray absorption ratio. DMR images after hyperbaric exposure showed calcification in the heads of long bones (humeri, femora, and tibiae) in mice. There were also significant changes in the X-ray absorption ratio in the heads. The accumulation of 99mTc-MDP was higher in all long-bone heads after hyperbaric exposure than before exposure. These results suggest that the hyperbaric environment promotes bone calcification. PMID- 9343863 TI - Cloth color preference under the influence of menstrual cycle. AB - The aim of this study is to know the effects of the menstrual cycle on cloth color preference in 10 normal menstruating subjects. They were instructed to choose a single one out of 41 cloth colors (24 x 52 cm), preferred by themselves, every five minutes from 06:30 h to 08:00 h under the ambient temperature (Ta) of 28 degrees C. A warmer cloth color was preferred in the luteal phase compared with the follicular phase. The findings that the warmer cloth color preference occurred in the luteal phase could probably be interpreted in terms of an establishment of a higher setpoint for core temperature. PMID- 9343864 TI - Development of a new algorithm for heat transfer equation in the human body and its applications. AB - A computer program has been developed for the numerical analysis of thermal conditions within the human body. In this study, a cylindrical model, which consists of internal multi-layers, is adapted for the segment of the human body. For the present concentric cylindrical model we developed a new numerical solution method using a linear combination of the modified Bessel functions. By using the present computer program the internal tissue temperatures, heat fluxes and blood temperatures of the thigh, crus and foot segments are calculated. This paper describes a new algorithm solving heat transfer equation in the human body and the example of calculated results of the combination of thigh, crus and foot segments. The calculated results show the local characteristics of each segment in the human body. PMID- 9343865 TI - The effects of a newly designed air mattress upon sleep and bed climate. AB - In our previous study, an air mattress with three series of air cells with inflation pressure, that was increased/decreased in time interval of 20 min, did not affect sleep quality and quantity, although the relative and absolute humidity inside the bedding was kept significantly higher than that of a Futon. The purpose of this study was to confirm the effects of a newly designed air mattress upon sleep and bed climate. In this newly designed air mattress, the cell series and time interval was reduced. Six healthy female volunteers, aged 18 22, served as subjects. The experiment was carried out under two conditions: using a regular Futon (Futon), and a newly-designed air mattress with the timer and pump activated (Airmat). The room temperature and relative humidity were controlled at 22-23 degrees C and RH 50-60%, respectively. The subjects' sleep was monitored by using an EEG machine and their skin temperatures and bed climates were also measured continuously. Subjective evaluations of bed comfort and sleep were obtained before and after the recording sessions. Sleep onset latency, wake after sleep onset and the sleep efficiency index showed no significant differences between the two conditions. A significant difference was observed in the bed climate of the waist area. The temperature of the waist was lower overall under the Airmat than the Futon, while relative humidity was higher under the Airmat. Absolute humidity also tended to be higher in the Airmat. Sleep evaluation and comfort sensation were good under both conditions. Although sleep was not disturbed and subjective sleep evaluation tended to be better in Airmat, our results indicate that changing the time intervals and cell series until this air mattress level is not effective in decreasing the bed climate humidity. PMID- 9343866 TI - Subcutaneous fat distribution of the abdomen and buttocks in Japanese women aged 20 to 58 years. AB - Subcutaneous fat is an essential element in shaping the body of human beings. In this research, skinfold thickness was measured specifically in 33 regions of the human body, including the abdomen and buttocks. Based on our measurements, the subcutaneous fat distribution was assessed for several age groups. The subjects were healthy Japanese women aged 20 to 58 years. Skinfold thickness was measured using the B-mode ultrasound methods, together with anthropometric measurement. A comparison was made between the following five age groups: early 20's, late 20's, 30's, 40's and 50's. The measured values for the early 20's group were used as the standard and the relationship between increase ratio of subcutaneous fat and age was studied. Through our research, we obtained data on the subcutaneous fat distribution in each age group. The largest change was observed between the ages of the early 20's and late 20's. The skinfold thickness measurements of the abdomen and buttocks was consistently around 10 mm for the early 20's, and increased up to 23.8 mm on the rear side section for the late 20's. This result indicates that the increase ratio varied depending on the part of body. Furthermore, the changes in skinfold thickness were different in specific parts of the abdomen and buttocks among different age groups. The difference in skinfold thickness between upper and lower sections of the abdomen also becomes more pronounced with age. Skinfold thickness increased significantly between the early 20's and late 20's. Among the body regions, measurements at the rear side showed the largest change with age; averaging 11.3 mm for the early 20's compared to 33.6 mm for the 50's. The subcutaneous fat distribution on the buttock also showed the differences with age, indicating changes in body shape. Using careful measurements of the abdomen and buttocks, subcutaneous fat distribution among each age group was determined as well as the variation in changes with the aging process. PMID- 9343867 TI - Tallal/Rice debate continues. PMID- 9343868 TI - Tallal/Rice debate continues. PMID- 9343869 TI - Body-mind-spirit responses. PMID- 9343870 TI - Body-mind-spirit responses. PMID- 9343871 TI - Body-mind-spirit responses. PMID- 9343872 TI - Body-mind-spirit responses. PMID- 9343873 TI - When is a difference too different? When it makes people uncomfortable.... PMID- 9343874 TI - Designing a future. PMID- 9343875 TI - Why specialty recognition? PMID- 9343876 TI - Linking up with telehealth. PMID- 9343877 TI - Knocking on the right doors. PMID- 9343878 TI - ASHA NOMS--a case study. PMID- 9343879 TI - Ahead of the curve. Improving service with speech-language pathology assistants. PMID- 9343880 TI - Practice versus evidence in diagnostic audiology. PMID- 9343881 TI - Late blooming or language problem? PMID- 9343882 TI - Telehealth and the Internet. PMID- 9343883 TI - Research and evidence-based decision making. PMID- 9343884 TI - O tell me the truth about evidence. PMID- 9343885 TI - Health and tuberculosis in Sydney's homeless. PMID- 9343886 TI - Surveillance for tuberculosis among residents of hostels for homeless men. AB - Tuberculosis has been recognised as an important health problem among homeless persons. The New South Wales tuberculosis screening program for residents of hostels for the homeless has been in operation for several years, but has not yet been evaluated. This study reviewed the performance of the tuberculosis surveillance program (which uses mobile chest x-ray screening) between 1989 and 1993 at the five major hostels for homeless men in the eastern Sydney area. Reports of the screening x-rays and records of subsequent follow-up examinations at chest clinics were examined; information on cases detected by the screening program was compared with notifications in the same population. Of 3555 residents screened during 23 visits, 506 (14.2 per cent) were found to have an abnormal chest x-ray. However, only two active cases of tuberculosis were diagnosed as a result of the screening program, while seven cases were notified on the basis of clinical presentation. About 50 per cent of those with an abnormal chest x-ray from the screening program were lost to follow-up. Possible reasons for loss to follow-up were: long delays in making chest clinic appointments; short-stay residents changing shelters without trace; and high prevalence of severe mental illness or organic brain syndrome among residents. Raising awareness of the disease among primary health care and welfare staff who work with homeless men may be a more effective approach to improving identification of cases of active tuberculosis in this population. PMID- 9343887 TI - Work-related hand and lower-arm injuries in New Zealand, 1979 to 1988. AB - The aim of this study was to describe the epidemiology of work-related hand and lower-arm injuries in New Zealand. Nonfatal hand and lower-arm injuries were identified from New Zealand's national database of hospital admissions for the period 1979 to 1988. Thirty-seven per cent (9714) of all such injuries (26,228) were work-related. Piercing and cutting instruments (38.5 per cent) and machinery (37.2 per cent) were the two most common agents of work-related hand and lower arm injury. Specific occupations in which the number of cases was high included meat workers (n = 1020, 3.3 per 1000 employees), carpenters (n = 548, 2.2 per 1000), machine operators (n = 450, 11.9 per 1000) and sawmill workers (n = 498, 7.7 per 1000). The injury rate for meat workers, carpenter-joiners, machine operators and sawmillers increased significantly over the 10-year study period. PMID- 9343888 TI - Risk-taking behaviours in a sample of New Zealand adolescents. AB - We surveyed the risk-taking behaviour of students aged 16 years and over in two New Zealand high schools, with a particular focus on road safety, substance use, sexual behaviour and personal safety. The questionnaire was completed by 471 students, a participation rate of 99 per cent. We found that seven out of 10 students who had ridden either a bicycle or motorcycle in the previous 12 months had not always worn a helmet; that 56 per cent had driven a car without a licence; and 23 per cent had been involved in a motor vehicle crash. A lifetime incidence of 63 per cent for cigarette smoking, 34 per cent for marijuana use and 78 per cent for alcohol use was found. Forty per cent of the students reported ever having sexual intercourse. During the previous 12 months, 49 per cent of these had not always used contraceptives and 61 per cent reported not always wearing condoms as protection for sexually transmitted diseases. Twenty-five per cent had physically harmed another person and 10 per cent reported carrying a weapon with the intent of harming someone else. This study shows that adolescents are willing to provide information on risk taking and that they are engaging in high levels of health-harming behaviour. Such information is important for designing health promotion programs to address adolescent risk taking. PMID- 9343890 TI - Inappropriate use of medications in the veteran community: how much do doctors and pharmacists contribute? AB - There is widespread inappropriate use of pharmaceuticals. Problems are with the prescribing and dispensing as well as with the taking of medicines. Levels of potentially inappropriate prescribing were estimated in the population of Australian veterans and war widows. This group tends to have multiple medical conditions and therefore to rely on medication therapy. These factors, and the average age of the population (72 years), increase the possibility of drug misadventures in this group. Despite the obvious hazards, a large number of high risk prescribing situations were detected. Although some of the problems can be reduced through changes in prescribing behaviour, the solution does not lie in the hands of doctors alone. PMID- 9343889 TI - Factors associated with falling in older Adelaide residents. AB - The aim of this study was to identify characteristics that predispose older residents of Adelaide to falling. Information collected in the baseline phase of the Australian Longitudinal Study of Ageing was used to draw cross-sectional comparisons between participants who reported having fallen on at least one occasion in the previous 12 months and those participants who reported not having fallen. The baseline cohort consisted of 1947 participants aged 70 years or more, of whom 550 (28 per cent) reported having fallen at least once in the previous year. Independent risk factors for falling were: age; having left school at an early age; a worsening of vision in recent years; and histories of Parkinson's disease, fractured hip, glaucoma, stroke (including transient ischaemic attack), corns or bunions, or arthritis. The findings regarding medical histories suggest some possible opportunities for reducing the risk of falls in the elderly by managing the symptoms and risk factors of underlying conditions such as stroke and loss of vision. PMID- 9343891 TI - Accuracy of ICD-9-CM codes in hospital morbidity data, Victoria: implications for public health research. AB - Hospital morbidity data in the form of International classification of diseases, 9th revision, clinical modification codes are often used for epidemiological studies and disease surveillance. We aimed to evaluate the reliability of the Victorian In-patient Minimum Database for use in epidemiological studies and disease surveillance. Data from 1993-94 were collected, as part of a coding audit of public hospitals in Victoria, from 7052 randomly selected records. The frequency of discrepancy in any coding field was 53 per cent, and of discrepancy in the principal diagnosis, 22 per cent. New Australian national diagnosis related group (ANDRG) codes were assigned as a result of discrepancy in 13.6 per cent of cases. Discrepancy rates increased with increasing rarity of ANDRG, from 50 per cent to 56 per cent. Predictors of change in ANDRG assignment were discrepancy in the principal diagnosis, ANDRG frequency of over 0.6 per cent, more than three diagnoses, medical ANDRGs, length of stay over five days and rural hospitals. Rates of any discrepancy increased from 36 per cent in patients with one diagnosis to 94 per cent in patients with 12 diagnoses. The discrepancy rates were consistent with those of other studies. Coding discrepancy is likely to be caused by universal difficulties associated with the coding of hospital records, rather than any unique local problems. The predictors of discrepancy suggest that more complex cases are more prone to coding discrepancy. In areas where the database is less reliable, use of a supplementary data source, such as link-age studies, would improve reliability. PMID- 9343892 TI - Trends in perinatal characteristics in South Australia, 1981 to 1994, by place of residence of mother. AB - We investigated differentials and time trends in perinatal mortality and perinatal risk factors by geographic area of residence in South Australia during 1981-1994, to assess whether sociodemographic inequalities had lessened. The areas analysed were Adelaide and the country region of South Australia, with Adelaide being divided by socioeconomic status into two ares. Subjects were 267,116 singleton births of at least 400 g birthweight (or at least 20 weeks' gestation) notified to the state's perinatal data collection. Year of birth, residential area, and interactions between year of birth and residential area were analysed as predictors of perinatal risk factors and deaths. There was a statistically significant decline in the perinatal death rate in all residential areas (mainly because of a decrease in neonatal deaths), which did not vary significantly by area. The frequency of low birthweight (< 2500 g) increased in the country areas and in the lower socioeconomic areas of Adelaide, but not in the higher socioeconomic areas. Although premature births increased in all areas, the increase was less pronounced for the higher socioeconomic areas of Adelaide. By comparison, although all areas showed an increase in the proportions of mothers aged 35 years or over, the increase was larger for the higher socioeconomic areas. Australia has a national policy of reducing social inequalities in health status. Perinatal mortality rates declined in Adelaide and country residential areas from 1981 to 1994. This trend is favourable, but from the relativities of these rates by residential area, there is not compelling evidence of a reduction in inequalities. PMID- 9343893 TI - Characteristics of infants receiving prompt first diphtheria-tetanus-pertussis immunisation in an infant cohort. AB - The Centers for Disease Control in the United States have stated that studies to determine factors associated with failure to receive the first recommended dose of diphtheria-tetanus-pertussis are required. We examined an infant cohort to identify family and infant characteristics predictive of prompt first immunisation, to document changes in prompt first immunisation rates over time and to identify reasons for immunisation delay. The study sample consisted of one fifth of live births in Tasmania at risk of sudden infant death syndrome. From 1 January 1988 to 31 December 1994, families of 8011 infants (83 per cent of eligible infants) participated in a telephone interview when the infants were a median postnatal age of 11 weeks and 3 days. Prompt immunisation was defined as the report by parents of diphtheria-tetanus-pertussis vaccination before a postnatal age of 10 weeks. The proportion of cohort infants promptly immunised increased (P < 0.0001) over time from 1988 to 1994. Prompt immunisation was associated with various characteristics of the infant and family. The proportion of infants promptly immunised decreased as birth order increased and as the interpregnancy interval between the index child and his or her immediately elder sibling decreased. After exclusion of infants not promptly immunised because of illness, birth order and interbirth interval remained significant predictors of prompt immunisation, suggesting that these factors are acting to increase immunisation delay through pathways unrelated to their potential effect on infant illness rates. PMID- 9343894 TI - Future directions for clinical practice guidelines: needs, lead agencies and potential dissemination strategies identified by Australian general practitioners. AB - There has been an increasing interest in developing clinical practice guidelines for general practitioners as a means of improving health outcomes. We conducted a survey of a national random sample of Australian general practitioners in May 1995 to determine their needs, preferred formats and dissemination strategies and to identify potential lead agencies for guidelines development. Of 373 eligible general practitioners, 286 (77 per cent) returned completed questionnaires. At least 50 per cent of respondents considered guidelines in angina, psychotic illness, skin cancer and attention deficit disorder as 'extremely' or 'very' useful. However, three other topics identified as areas for future guidelines development in Better health outcomes for Australians were so rated by less than half of the general practitioners surveyed. The Australian Cancer Society and the Australian Medical Association outranked nine other organisations in terms of credibility in guidelines development. Innovative formats, including computerised medical records or text, were not highly rated, consistent with our finding that only 27 respondents (9 per cent) had Internet access. Strategies nominated as likely to increase the adoption of a guideline included a personal visit by a trained nurse, a lecture about its content or a Medicare rebate being available when a patient was managed in accordance with the guideline. Public health practitioners and nominated lead agencies are encouraged to respond to these findings and recognise potential strategies to enhance the effective dissemination of guidelines. Interventional studies are required to demonstrate and allow understanding of changes in clinical practice attributable to guidelines. PMID- 9343895 TI - Initiation rate and duration of breast-feeding in the Melbourne aboriginal community. AB - Breast-feeding is important for child health and helps to protect the child against infections. The objective of this study was to determine baseline breast feeding rates in the Melbourne Aboriginal community prior to a breast-feeding promotion project. A brief questionnaire was administered to 116 mothers of infants up to two years of age with a Melbourne metropolitan address. During their pregnancies, 99 (85.3 per cent) of the women had planned to breast-feed, and 98 (84.5 per cent, 95 per cent confidence interval (CI) 77.9 to 91.1 per cent) initiated breast-feeding. However, 10 (8.6 per cent) stopped within the first week, and seven (6 per cent) more stopped within the first four weeks. Only 50 per cent of the babies (CI 40.9 to 59.1 per cent) were still being breast-fed at three months of age and 32 per cent were still being breast-fed at six months of age (CI 23.5 to 40.5 per cent). Younger mothers were less likely to choose to breast-feed (73 per cent) than women 20 years and over (87 per cent) and were also more likely to stop breast-feeding within three months. A total of 45 (51.1 per cent) of the babies received food other than breast milk or formula earlier than the recommended minimum age of four months. These results are similar to those for the general Victorian population. They show that while most Aboriginal women choose to breast-feed, many cease breast-feeding before they had intended. PMID- 9343896 TI - Influences on infant-feeding beliefs and practices in an urban aboriginal community. AB - The Victorian Aboriginal Health Service initiated a project to increase breast feeding rates in the Melbourne Aboriginal community. The results of focus-group discussions on infant-feeding experiences and beliefs provided a wealth of information for the design of appropriate interventions. Most women wanted and expected to breast-feed. Some chose artificial feeding because of embarrassment, a belief that it is as good as breast-feeding, or perceptions that breast-feeding is painful and inconvenient. The most common reasons that women stopped breast feeding were sore nipples, worries about their supply of milk and tiredness. Lack of knowledge, hospital practices, lack of support and appropriate advice, and lack of confidence and self-esteem contributed to these problems. Disruption of the passing on of knowledge of healthy infant-feeding practices between generations is another cultural loss suffered by Aboriginal communities. Efforts to restore traditional rates of breast-feeding need to be under Aboriginal control and to take account of these influences. PMID- 9343897 TI - Environmental health conditions in remote and rural aboriginal communities in western Australia. AB - During 1994-1995 environmental health conditions of about 13,760 persons in 155 remote and rural Aboriginal communities in 20 local shires in Western Australia (WA) were surveyed. A semiquantitative questionnaire sought data about the communities and their services, including water supplies, power, sanitation and disposal of solid and liquid waste; a separate section dealt with conditions of individual dwellings. Data were recorded by experienced local workers. Thirty five communities considered to have the worst conditions were evaluated on-site by a team of senior personnel in mid-1995. Environmental health problems were prevalent and often serious: over one-third of the communities had water supply or sanitation problems and 70 per cent had housing problems, with overcrowding and substandard housing being commonplace. Thirty-six per cent had difficulties with waste water disposal, 37 per cent had no rubbish disposal, and in others, the methods of disposal were often inadequate; pests were problems in 44 per cent of communities and the hygiene and maintenance of communal toilets was unacceptable in 25 per cent. Seventy-two per cent had no on-site environmental health worker and 44 per cent had no on-site or visiting medical, nursing or health worker personnel. An action plan was developed and the highest-priority communities were targeted in a program of major works (for example, housing, drainage and sewerage) and minor works, which have been commenced. The remote area environmental health workers' program is being expanded. Increased intersectoral collaboration and enhanced community involvement in decision making have occurred as a result of this work. PMID- 9343898 TI - A community-based approach to the control of sexually transmitted diseases in the Northern Territory. AB - A program to control sexually transmitted diseases (STDs) was undertaken during a Men's Health Week in a remote Aboriginal community in Western Arnhem Land, Northern Territory. A total of 151 men aged 13 years and over who attended over a five-day period underwent a full physical examination, and first-void urine specimens were tested for the presence of leukocytes, chlamydia (by enzyme immunoassay antigen detection) and gonorrhoea (by culture and antigen detection). Blood was taken for syphilis serology from all patients and for human immunodeficiency virus (HIV) from patients with a proven STD or at the patient's request. Consent for testing was obtained from all participants. Patients with a positive urinary leukocyte test or symptoms were offered urethral swab investigations and treated empirically according to a set protocol. Patients with STDs detected by subsequent laboratory investigations were followed up and treated. The overall prevalence of one or more of syphilis, gonorrhoea or chlamydia was 17.4 per cent. No men presented with genitourinary symptoms and none was HIV-infected. In this population, STDs were an important cause of morbidity, and a community-based approach was adopted to identify infected persons. The use of urine for the detection of gonorrhoea and chlamydia was highly acceptable. Although not used in this study, polymerase chain reaction and ligase chain reaction technology will facilitate similar activities in the future. PMID- 9343899 TI - Risk factors for aboriginal low birthweight, intrauterine growth retardation and preterm birth in the Darwin Health Region. AB - Risk factors for Aboriginal low birthweight (< 2500 g), preterm birth (< 37 weeks' gestation) and intrauterine growth retardation (under the tenth percentile of Australian birthweights for gestational age) were examined in 503 live-born singletons recorded as born to an Aboriginal mother and routinely delivered at the Royal Darwin Hospital between January 1987 and March 1990. Infants born to mothers with body mass index less than 18.5 kg/m2 had five times the risk of having low birthweight and 2.5 times the risk of intrauterine growth retardation. Population-attributable risk percentages suggest that 28 per cent of low birthweight and 15 per cent of growth retardation could be attributed to maternal malnutrition. Risk percentages for maternal smoking of more than half a packet of cigarettes a day were 18 per cent for low birthweight and 10 per cent for growth retardation. For growth retardation, 18 per cent could be attributed to a maternal age under 20 years. Risk factors for preterm birth were predominantly obstetric: the population-attributable risk percentage for pregnancy-induced hypertension was 26 per cent and for other obstetric conditions was 16 per cent. For Aboriginal births in the Darwin Health Region, maternal malnutrition and smoking are key elements in the prevention of low birthweight and intrauterine growth retardation. Teenage pregnancy is an important risk for intrauterine growth retardation, and pregnancy-induced hypertension is a risk for preterm birth. PMID- 9343901 TI - The food-service industry, dietary guidelines and change. AB - The influence of the food-service industry on compliance with the Australian dietary guidelines was investigated through three separate methods of data collection and analysis: a telephone survey of 1683 randomly selected Brisbane residents; telephone interviews with 69 food-service-industry operators and 10 face-to-face interviews with key stakeholders in industry and government. Nearly 40 per cent of respondents had consumed foods prepared by the food-service industry at least once on the day before the interview, mainly from restaurants, cafes and takeaway shops, in the form of fast-food or snacks. Consumption of these foods declined with age. Those consuming foods prepared by the food-service industry ate significantly less fruit, vegetables and dairy food and were therefore less likely to comply with the dietary guidelines. Outcomes from interviews with operators in the food-service industry show that food choices offered to consumers were the result of a dynamic interaction between consumer demand and operators' own tastes and perceptions of food quality. Key informant interviews show that public health nutrition programs will have limited effect without supportive environmental changes in the food-service industry supply. An effective means of increasing the likelihood of compliance with the Australian dietary guidelines will be to encourage food suppliers in ways that address their core business concerns simultaneously with the goals of health professionals. PMID- 9343900 TI - Senior school students' experiences and opinions of school-based HIV-AIDS education. AB - This article reports the findings from the second part of a two-stage study that used both qualitative and quantitative methods to investigate the communication context of school-based HIV-AIDS education in state secondary schools in metropolitan and rural areas of New South Wales. The quantitative data are here described, focusing on a sample of 1005 Year 12 students' responses to a self administered questionnaire. The data suggest that the students strongly supported the general idea of school-based HIV-AIDS education, but found current offerings lacking in several respects. Students identified a strong need for information about how HIV and AIDS affect the body, for more information about sexually transmissible diseases other than HIV-AIDS, for people with HIV themselves and experts in the field to provide education sessions, and for more small-group discussions. Rural students and those students from schools located in the outer western suburbs of Sydney in particular reported that they had insufficient access to the modes of information that they most preferred. There were some important differences between the responses of female and male students and between the responses of students from different ethnic groups, suggesting that these factors also need acknowledgment when school-based programs are designed for young people. PMID- 9343902 TI - A self-administered questionnaire for detection of unrecognised coronary heart disease. AB - On an individual and a population basis, an increased incidence of coronary heart disease is associated with classical cardiovascular risk factors, but many cases occur in people not identified as at high risk. Conversely, many people at high statistical risk do not develop coronary disease. We used a questionnaire to identify unrecognised coronary heart disease in people attending large-scale health survey centres. Participants were required to report the presence and characteristics of any chest pain. Those returning responses consistent with myocardial ischaemia were offered treadmill exercise ECG tests. Over 18 months, 4070 questionnaires were returned. Of 475 respondents offered testing, 229 (198 male, 131 female) accepted. Thirty-two subjects (15 male, 17 female: a detection rate of 13.9 per cent of those assessed as likely on questionnaire, or 0.8 per cent of all respondents) had results consistent with significant coronary heart disease. Follow-up was available in 30 cases. There was no difference in classical risk-factor distribution (including multivariate risk percentiles: 42.4 (male) and 46.7 (female)) between those newly diagnosed with coronary heart disease and their community counterparts. More women than men were identified as suffering from unrecognised coronary heart disease, with a preponderance of younger women. Cost per case identified was A$1220. Screening by self administered questionnaire is a useful and relatively cost-effective means of identifying unrecognised coronary heart disease. PMID- 9343903 TI - Review of evidence on fluoridation. PMID- 9343904 TI - Heterotopic transplantation as a model to study the regulation of spermatogenesis; some histomorphological considerations about sperm decline in man. AB - A novel animal model (syngeneic neonatal testicular graft transplanted under the skin of the outer ear in adult inbred Fischer rats that had been castrated, hypophysectomised, and/or subjected to hormonal replacement therapy) was developed to study regulation of spermatogenesis and Sertoli cell number. Given that Sertoli cell number and testicular size are important in determining spermatozoan production rates, this model was first used to study Sertoli cell proliferation, testicular size, and establishment of germ cells. The specific objectives were to determine the developmental pattern of Sertoli cell numbers in transplanted testes and the effect of number of testes transplanted, sex of hosts, pituitary hormonal removal, and replacement on Sertoli cell number, hormonal status of the host, and establishment of germ cells. A few tubules had complete spermatogenesis at 90 days posttransplantation, indicating that Sertoli cells in some of these tubules were functional. Leydig cell structure appeared to be normal, but the density of these interstitial cells was greater than that in testes of intact rats. Although the weight of the seminal vesicles and prostate were maintained in the castrated host with transplants, both serum FSH and LH concentrations were higher than intact control rats. Leukocytic infiltration of testes was not observed in intact rats or in rats receiving neonatal testes. Although transplanted testes showed a delay in reaching the plateau value for Sertoli cell number per testis and although the value reached was lower than in intact testes, the developmental pattern of Sertoli cell proliferation (early division of cells followed by stabilized number of cells) in transplanted testes was similar to that in intact rats. Hypophysectomy reduced the growth of testicular grafts, and hormonal replacement via retransplantation to pituitary intact hosts enhances Sertoli cell proliferation and testicular growth. When two on four testes were transplanted into castrated males or ovariectomized female hosts for 65 days, there was no difference in the graft weights or Sertoli cell numbers between sexes of hosts. Four transplanted testes per rat produced more total testicular parenchyma and a greater number of Sertoli cells per testis than did two testes regardless of sex of the host. Transplantation of six or eight testes produced more total Sertoli cells/host than that found in testes of intact rats. Using hormonal therapy in hypophysectomized hosts, the testicular parenchymal weight was greater for pituitary-intact hosts and FSH-LH combination than the control media. There was no statistically significant difference among the media control, LH, FSH, and GH. This testicular transplant model has shown that the period of Sertoli cell proliferation can be delayed by hypophysectomy, that Sertoli cell number can be influenced by endogenous and exogenous hormones, and that a major component in regulation of testicular size is at the level of the testis. Hence, this model should facilitate study of experimental endocrine manipulation control and potential experimental intervention to increase Sertoli cell number, testicular size, and spermatogenesis. Regarding human sperm count decline in recent years, there appears to be no significant decline in Sertoli cell number or spermatogenic potential in a group of North American men. However, there was a significant decline in volume/man of Leydig cells and volume/man of Leydig cell cytoplasm. PMID- 9343905 TI - [Precocious menopause after antimitotic therapy. Is in vitro fertilization with oocyte donation the only appropriate response?]. AB - Advances in antimitotic treatments have improved the prognosis of cancer in young subjects. The resulting increase in life expectancy raises the question of the subject's future fertility, a question that should be posed before beginning any anticancer therapy which could lead to a gonadal failure. If oocyte donation remains the alternative indication proposed for these patients desiring a child, it is important to assess the tissue alterations in the uterus, to verify its vascularization under suitable treatment and appreciate the other alternative directions. PMID- 9343906 TI - A critique of the case for privatizing Social Security. AB - This article presents and then critiques arguments made by those advocating the privatization of Social Security. It refutes the argument that baby boomers will find themselves without Social Security pensions unless fundamental changes are made. It questions the claims that privatization would increase economic growth, reduce the federal deficit, make the nation more competitive in the global economy, protect workers against payroll tax increases, protect boomers against pension benefit cuts, and increase confidence in Social Security. It argues that for low-wage workers, returns on contributions would probably decrease and future benefits would become politically more vulnerable. PMID- 9343907 TI - Top management team characteristics and innovation in nursing homes. AB - This study examines how the demographic characteristics of the top management team in 236 nursing homes can affect the adoption of innovations. The computerization of the Minimum Data Set (MDS) is the innovation we examine, and tenure, education, and involvement in a professional society are the demographic characteristics investigated. Controlling for 10 organizational and environmental factors, the results are generally significant for each of these demographic factors. However, the results for top managers of nonchain nursing homes show a greater association between these demographic factors and innovation than the results for top managers of nursing homes belonging to a chain. We discuss these results in terms of their significance for innovation research, nursing homes, and top management. PMID- 9343908 TI - Psychometric and psychodynamic correlates of first memories in younger and older adults. AB - We conducted two studies that identified some of the psychometric and psychodynamic correlates of the first memories of younger and older adults. Collectively, these studies revealed that: (a) performance on the Weschler Adult Intelligence Scale-Revised (WAIS-R) vocabulary and digit-span backwards tasks was negatively related to the age of younger and older adults' first memories, (b) performance on the death preparation subscale of the Reminiscence Function Scale was inversely related to age of older adults' first memories, but positively related to the age of younger adults' first memories, and (c) the use of internalizing defenses as measured by the Defense Mechanism Inventory was predictive of the age of younger, but not older, adults' first memories. The implications of these data for theories of infantile amnesia and life review are discussed. PMID- 9343909 TI - Morbidity and comorbidity among Great Lakes American Indians: predictors of functional ability. AB - This article explores patterns of morbidity and comorbidity and their ability to predict functional disability among American Indian elders, using data from a sample of urban, rural off-reservation, and reservation Great Lakes American Indians age 55 and older. Higher rates are reported of a number of chronic illnesses than found in overall samples of U.S. elders. Results of multiple regression analyses predicting Instrumental Activities of Daily Living (IADLs) and Activities of Daily Living (ADLs) show age to be a consistent predictor of functional disabilities: The CMI (Index of Comorbidity) was found to be a more useful predictor of functional disability than was the simple summation of the number of chronic illnesses. PMID- 9343910 TI - The treatment decision-making process: age differences in a sample of women recently diagnosed with nonrecurrent, early-stage breast cancer. AB - This research utilizes retrospective, self-report data collected from a nonprobability sample of women recently diagnosed with nonrecurrent, early-stage breast cancer to better understand how the treatment decision-making process varies with patient age. Three important areas--context, decision-making style, and influencing factors--are examined using bivariate and multivariate analyses. Findings indicate that although patients recalled similar contextual attributes, they reported attitudes, behavior, and considerations that differed by age. Older women were less likely than their younger counterparts to have desired participation in therapy selection, sought out medical information, or considered the possibility of recurrence when making treatment decisions. PMID- 9343911 TI - Adherence to antihypertensive medications across the life span. AB - Although treatment for hypertension is readily available, poor control of hypertension is a major health problem frequently manifested in late life. Researchers believe that one of the major causes of uncontrolled hypertension is failure to take medication as directed. In this preliminary study, the medication taking behaviors of 48 adults diagnosed with hypertension, ranging in age from 35 to 87, were recorded for 2 months with credit card-sized bar-code scanners. The social-cognitive model (Park, 1992) for understanding medication adherence, which proposes that medication adherence is governed by both beliefs and cognitive factors, was used as a basis for this research. Therefore, measures of health behaviors, attitudes about health and medication taking, and cognitive function were recorded, as well as blood pressure readings. The main findings were that (a) the oldest-old and groups of middle-aged adults were the most nonadherent, whereas the young-old were more likely to adhere than the other age groups; (b) high blood pressure readings predicted adherence to antihypertensive medications; and (c) medication beliefs influenced adherence in some situations. PMID- 9343912 TI - The relationship between vision impairment and the assessment of disruptive behaviors among nursing home residents. AB - This study examines the relationship between vision impairment, defined as best corrected distance acuity, and disruptive behaviors among nursing home residents (N = 89). All data were collected from nursing home records. Vision impairment was significantly related on the bivariate level to the disruptive behavior index (r = .21; p < .05). Hierarchical regression analyses, with disruptive behaviors as the criterion and age and comorbid conditions as covariates, indicate that vision status is a significant independent contributor to disruptive behaviors among long-term care residents. Several interpretations for this observed relationship are discussed, as are implications for nursing home services and future research. PMID- 9343914 TI - Everyday ethics in dementia day care: narratives of crossing the line. AB - The purpose of this study was to gain understanding of the ethical aspects of the experience of providing day care to people with dementia. Telephone interviews were conducted to elicit phenomenological narratives of satisfying and dissatisfying experiences from staff members of a state-wide random sample of dementia day care facilities. The analysis was guided by the concept of situated ethics. Findings reveal that ethical challenges of dementia day care are embodied in the everyday incidents when participants, staff, or family members "cross the line" of acceptable behavior. An ethical hierarchy of staff responses ranges from benign manipulation to termination of day care. These findings help us understand the situated ethics of dementia day care and heighten our sensitivity to the lived experience of dementia day care staff. PMID- 9343913 TI - Prevalence of behavior disorder and disturbance to family and staff in a sample of adult day health care clients. AB - Latent class-derived prevalence estimates of behavior disorder are provided for adult day health care (ADHC) clients; informal and formal caregivers reported 11% and 14%, respectively, of these clients as engaging in severe disturbed behavior (95% confidence intervals across sources are from 7% to 18%). The prevalences, estimated for informal and formal caregivers respectively, were 12% and 16% for affective disorder, 15% and 18% for cognitive disorders, 16% and 13% for verbal vocal agitation, and 6% and 8% for socially inappropriate behavior. These rates can be contrasted with those of the institutional population which, while higher, overlap with the distribution of behavior disorder for ADHC community residents. The degree of reported disturbance to family and staff was similar across items. PMID- 9343915 TI - Grandmother involvement in child caregiving in an urban community. AB - In a community-defined, epidemiologic sample in East Baltimore, we examined grandmothers' rates of co-residence and their involvement in four parenting activities. Co-residence rates exceeded the national average. Six types of family households with grandmothers were identified, and their frequency varied by race. Neither grandmother age nor employment was associated with grandmothers' parenting involvement, although family structure was. Grandmothers who were the sole parent (21%) or co-parent with a grandfather (6.5%) were most involved in child care and had the fewest number of helpers. Grandmothers living with single mothers (41%) were the next most involved, while grandmothers in mother/father households (9%) were least involved. PMID- 9343916 TI - Care-related decision-making satisfaction and caregiver well-being in families caring for older members. AB - Families provide care and make decisions regarding care within the context of specific structural characteristics and the broader family environment. This research analyzes data from 244 adult child and spouse caregiver interviews. It examines the impact of structural variables (e.g., caregiver type, elder impairment) and family environment (adaptability, conflict, cohesion) on satisfaction with care-related decision making and caregiver well-being. Regression analysis results indicate that aspects of family environment such as adaptability and conflict are the best predictors of decision-making satisfaction are the best predictors of caregiver depression. PMID- 9343917 TI - Health and well-being of spouse caregivers and the widowed. AB - The appropriateness of the stress-coping and life transitions models for late life experiences was assessed by examining psychosocial correlates of spouse caregiving in association with widowhood. Data were from the Health Status of Older People (HSOP) survey of 1,000 people aged 65 and over living in the community in Melbourne, Australia. Caregiving was found to be a usual precursor to widowhood and may have prepared older people for widowhood. However, these two life experiences had different consequences. While widowhood appears to be the more severe transition, caring for a spouse often seriously threatens the meaningfulness of life. PMID- 9343918 TI - Management of the patient with disruptive vocalization. AB - Disruptive vocalization (DV) is a common problem in the management of cognitively and physically impaired older people. This article reports the results of a consensus meeting convened to provide guidelines for clinicians and recommendations for researchers in this difficult and little-studied behavioral problem. DV arises largely in people with cognitive impairment and generally reflects an underlying need or discomfort. A variety of factors can precipitate and aggravate DV; the key to management is appropriate identification of all possible factors and development of an individualized treatment plan. PMID- 9343919 TI - Project REACH: a program to train community-based lay health educators. AB - The major purpose of this project was to test the feasibility of recruiting and training volunteers as lay health educators who could coordinate and reinforce the educational efforts of health care providers. A committee of health care professionals designed a 16-hour program. Twenty-five volunteers from T1 religious institutions and 4 retirement communities completed the 8-week program. The program was successful in identifying, recruiting, and training volunteers from racially and religiously diverse institutions. Favorable outcomes included participants' satisfaction and success in organizing numerous educational and screening programs in their communities. PMID- 9343920 TI - An intergenerational program for persons with dementia using Montessori methods. AB - An intergenerational program bringing together older adults with dementia and preschool children in one-on-one interactions is described. Montessori activities, which have strong ties to physical and occupational therapy, as well as to theories of developmental and cognitive psychology, are used as the context for these interactions. Our experience indicates that older adults with dementia can still serve as effective mentors and teachers to children in an appropriately structured setting. PMID- 9343921 TI - Is the role of fatty acids only to provide membrane-anchor in fatty acylated proteins? AB - A large number of proteins on the eukaryotic cell surface that play an important role in cellular metabolism are covalently modified with fatty acids like palmitic and myristic acids. While some of these proteins have transmembrane spanning segments, many others do not. Early hypothesis was that this co or posttranslational modification helped in membrane-association and the fatty acyl chain provided a stable membrane anchor. We have investigated the structure of peptides with these modifications and also their interaction with membranes. Our results indicate that the fatty acylated peptides, especially when the peptide segment is not hydrophobic, do not have strong affinity for membranes. The recent observations about the dynamic nature of fatty acid acylation as well as the importance of protein-protein interactions for function in fatty-acylated proteins suggest that membrane-association may involve factors other than only the fatty acid, either myristic or palmitic. Revised models depicting the possible role of fatty acids in modulating protein-protein interaction and their dynamics is presented. PMID- 9343923 TI - Role of hydrophobic effect and surface charge in surfactant-DNA association. AB - Ethidium bromide is one of the best known DNA intercalator. Upon intercalation inside DNA, the fluorescence due to ethidium bromide gets enhanced by many orders of magnitude. In this paper, we employed ethidium bromide as a probe for studying surfactant-DNA complexation using fluorescence spectroscopy and agarose gel electrophoresis. Surfactants of different charge types and chain lengths were used and the results were compared with that of the related small organic cations or salts under comparable conditions. The cationic surfactants induced destabilization of the ethidium bromide-DNA complex at concentrations in orders of magnitude lower than that of the small organic cations or salts. In contrast however, the anionic surfactants failed to promote any such destabilization of probe-DNA complex. DNA loses its ethidium bromide stainability in the presence of high concentration of cationic surfactant aggregates as revealed from agarose gel electrophoresis experiments. Inclusion of surfactants and other additives into the DNA generally enhanced the DNA double-strand to single strand transition melting temperatures by a few degrees, in a concentration-dependent manner and at high surfactant concentration melting profiles got broadened. PMID- 9343922 TI - Mycoplasma in the spotlight of AIDS: partial biochemical characterization of mycoplasma-derived components inducing TNF alpha secretion and blast transformation. AB - We have previously demonstrated that the AIDS-associated Mycoplasma fermentans as well as Mycoplasma capricolum membranes activated bone marrow macrophages to secrete tumor necrosis factor alpha (TNF alpha) and induce blast transformation of splenic lymphocytes. Herein, we show that the membrane component of Mycoplasma capricolum capable of inducing TNF alpha secretion is a hydrophobic protein. This is supported by our findings that the TNF alpha inducing activity was eluted by a phenyl-Sepharose column in a peak distinct from bulk membrane lipids. The hydrophobic nature of the protein is indicated by the activity of the "hydrophobic protein" fraction of the membranes, and the pattern of elution obtained by the phenyl-Sepharose column. Fractionation of the M. capricolum membranes, solubilized by CHAPS (3-[(3-cholamidopropyl)-dimethylammoniol]1 propane sulfate) on a gel filtration column revealed a major peak of TNF alpha inducing activity of about 75,000 daltons, and a minor peak of about 55,000 daltons. The mitogenic activity, though spread throughout the column, peaked in the same fractions as the TNF alpha inducing activity. Both activities co-eluted by the phenyl-Sepharose column as well. However, the mitogenic activity of the membranes was much more resistant to elevated temperatures and extreme pH treatment. PMID- 9343924 TI - A novel 200 kDa plasma membrane glycoprotein with high basal tyrosine kinase activity in tumour cells. AB - We have detected a tyrosine phosphorylated 200 kDa glycoprotein (gp200) on the surface of two tumour cells of neural origin, namely A1235 glioma and A172 glioblastoma. gp200 (polypeptide mass of 165-170 kDa) has all the structural features of a growth factor receptor and it appears to display high basal tyrosine kinase activity, a characteristic associated with transforming proteins. Another interesting feature of gp200 is that it is immunologically highly related to the EGF receptor (polypeptide mass of 135 kDa), a member of the erb-B family of proteins; however, it lacks EGF binding activity. gp200 also differs from all other EGF-receptor-related oncogenic proteins, namely erb-B-2, erb-B-3 and erb-B 4 gene products, and hence appears to be yet another member of the erb-B family of proteins. This is further strengthened by the fact that both gp200 and the EGF receptor are also recognized by a conformation-specific anti-peptide antibody to the cytoplasmic domain of the beta-type PDGF receptor. In the EGF- and the PDGF receptors, this peptide epitope is cryptic and receptor phosphorylation unmasks this site [Panneerselvam K, Reitz H, Khan S A, and Bishayee S (1995) J Biol Chem 270, 7975-7979] indicating that this epitope might be important in biological message transmission. In this context, the expression of a novel EGF-receptor related 200 kDa protein with high basal kinase activity in certain tumour cells of neural origin and the fact that it contains an important peptide epitope suggest its possible role in normal and abnormal cell growth. PMID- 9343925 TI - Intracellular signalling: phosphatidylinositol lipid metabolism in cancer cells. AB - Evidence for heightened capacity for signal transduction in rat hepatoma as well as in human breast and ovarian carcinoma cells as reflected by coordinate increases in PI kinase and PIP kinase in the PI phosphorylation sequence leading to the production of second messengers IP3 and DAG is shown. The linkage of signal transduction enzymes with malignant growth is also seen as MDA-MB- 435 human breast carcinoma or ovarian OVCAR-5 cells express their proliferative capacity in tissue culture in the log phase. In both cases, quercetin inhibit cell proliferation with a decline in PI kinase activity and IP3 levels preceding the growth inhibition seen with quercetin. The elevated steady state activities of PI and PIP kinase indicate a metabolic up-regulation in signal transduction capacity of cancer cells which is down-regulated by quercetin. Since the gain in function manifested in the over-expressed capacity for signal transduction confers selective growth advantage to cancer cells, increased activities of PI and PIP kinases may be considered as sensitive targets for cancer chemotherapy. The potential of quercetin as an interceptor of intracellular signal transduction mechanisms needs to be explored. PMID- 9343926 TI - Interaction of SH3 domain of Hck tyrosine kinase with cellular proteins containing proline-rich regions: evidence for modulation by unique domain. AB - The Hck tyrosine kinase, a member of Src family, is predominantly expressed in myeloid cells. In this report we have analyzed interaction of cellular proteins with Src homology 3 (SH3) domain of Hck. For this purpose we used various GST-Hck fusion proteins comprising a part of unique region, complete unique region and/or complete SH3 domain of Hck, and glutathione S-transferase (GST). When these fusion proteins (or GST), immobilized on glutathione-agarose beads were incubated with [35S] methionine labelled cell extracts, multiple proteins which interact specifically with SH3 domain of Hck were detected by SDS-PAGE followed by autoradiography. The Hck interacting proteins could also be detected by a tandem blot binding assay in which the blot was incubated with purified fusion protein (or GST) and then the interacting proteins were identified by using antibody against GST. When a part of or complete unique domain was present along with SH3 domain, the interaction of some specific proteins was reduced several fold. These results raise the possibility of unique domain altering the properties of SH3 domain, thus modulating or restricting the interaction of SH3 domain with specific cellular proteins. This modulatory effect of unique domain was localized to 28 amino acids upstream of SH3 domain. SH3 interacting proteins were associated with serine/threonine and tyrosine kinase activities towards exogenous substrates. Most of the SH3 binding proteins were soluble in Triton X-100. Differentiation of promyelocytic leukemia cell line HL-60 into macrophage like cells resulted in appearance of novel SH3 binding proteins. Hck was detected in the eluate of WGA-Sepharose column, suggesting that it interacts with WGA binding glycoprotein (s). A rat spleen cDNA library was screened for the SH3 binding proteins by protein interaction cloning. Sequence analysis of the clones showed the presence of proline rich regions containing PPXP motifs. PMID- 9343927 TI - Membrane guanylate cyclase signal transduction system. AB - The suspect role of the receptor-mediated cyclic GMP signaling pathway was dispelled by the discovery of a membrane guanylate cyclase that was also an atrial natriuretic factor receptor. It is now established that the membrane guanylate cyclase transduction system is linked to the signaling of natriuretic factors, guanylin/enterotoxin, and emerging evidence suggests that a new neural tissue-specific subfamily of membrane guanylate cyclases exists whose mechanism of signal transduction is different from those of the other membrane cyclases. This review will briefly discuss the fascinating, albeit turbulent, history of this signal transduction field, which will be followed by its current status and finally the direction it is heading. PMID- 9343928 TI - Specific inhibition of insulin receptor dephosphorylation by a synthetic dodecapeptide containing sulfotyrosyl residues as phosphotyrosyl mimetic. AB - We have synthesized a tris-sulfotyrosyl dodecapeptide (3S-peptide-I) that corresponds to the major autophosphorylation domain within the insulin receptor beta-subunit and showed that it potently inhibited insulin receptor dephosphorylation by protein tyrosine phosphatases (PTPases) in vitro. 3S-peptide I also inhibited tyrosine dephosphorylation of a synthetic peptide by the recombinant PTPase PTP-1B, indicating that 3S-peptide-I interacts directly with PTPase, causing its inactivation. The peptide had no effect on the activity of serine/threonine phosphatases, PP-1 and PP-2A, or alkaline phosphatase. Furthermore, we found that the introduction of a N-stearyl derivative of 3S peptide-I in CHO/HIRc cells caused a significant increase in insulin-stimulated phosphorylation of the insulin receptor. In contrast, ligand-stimulated phosphorylation of epidermal growth factor (EGF) receptor in CHO cells overexpressing EGF receptors was not affected by the presence of N-stearyl-3S peptide-I. These data suggest that by inhibiting dephosphorylation of the insulin receptor in intact cells, 3S-peptide-I may specifically enhance insulin signalling. PMID- 9343929 TI - Oxidized low density lipoproteins and lactosylceramide both stimulate the expression of proliferating cell nuclear antigen and the proliferation of aortic smooth muscle cells. AB - We have previously shown that oxidized low density lipoproteins (Ox-LDL) at low concentrations (10 micrograms/ml) via activating a UDP-galactose: glucosylceramide, beta 1-->4 galactosyl-transferase (GalT-2) and producing lactosylceramide can stimulate the proliferation of aortic smooth muscle cells. In this report, we present evidence that Ox-LDL and LacCer, both can induce the expression of proliferating cell nuclear antigen (cyclin). Ox-LDL and LacCer both exerted a time-dependent stimulation of cyclin expression. Maximum increase (3 fold) in cyclin expression occurred between 30-120 min after Ox-LDL/LacCer addition and decreased thereafter. D-threo-l-phenyldecanoylamino-3-morpholino-1 propanol (D-PDMP), an inhibitor of GalT-2, inhibited cell proliferation as well as cyclin expression. This inhibitor also abrogated the Ox-LDL mediated expression of proliferating cell nuclear antigen (cyclin). In contrast, the L enantiomer of PDMP (L-PDMP) stimulated the expression of cyclin and augmented the Ox-LDL mediated expression of cyclin in these cells. Maximum increase in the expression of cyclin occurred with 20 mumole of L-PDMP and 10 micrograms of Ox LDL. This overall pattern of Ox-LDL and LacCer mediated regulation is similar to that of the c-fos protooncogenes reported previously by us. We hypothesize that the early induction of GalT-2 may serve as an "Immediate early gene" that plays a role in the signalling cascade by LacCer and involves the kinase c-fos induction and subsequent expression of cyclins. Thus, GalT-2 may play a role in the proliferative response in aortic smooth muscle cells by Ox-LDL. PMID- 9343930 TI - Differential binding properties of Gal/GalNAc specific lectins available for characterization of glycoreceptors. AB - Differentiating the binding properties of applied lectins should facilitate the selection of lectins for characterization of glycoreceptors on the cell surface. Based on the binding specificities studied by inhibition assays of lectin-glycan interactions, over twenty Gal and/or GalNAc specific lectins have been divided into eight groups according to their specificity for structural units (lectin determinants), which are the disaccharide as all or part of the determinants and of GalNAc alpha 1-->Ser (Thr) of the peptide chain. A scheme of codes for lectin determinants is illustrated as follows: (1) F (GalNAc alpha 1-->3GalNAc), Forssman specific disaccharide--Dolichos biflorus (DBL), Helix pomatia (HPL) and Wistaria floribunda (WFL) lectins. (2) A (GalNAc alpha 1-->3 Gal), blood group A specific disaccharide--Codium fragile subspecies tomentosoides (CFT), Soy bean (SBL), Vicia villosa-A4 (VVL-A4), and Wistaria floribunda (WFL) lectins. (3) Tn (GalNAc alpha 1-->Ser (Thr) of the protein core)--Vicia villosa B4 (VVL-B4), Salvia sclarea (SSL), Maclura pomifera (MPL), Bauhinia purpurea alba (BPL) and Artocarpus integrifolia (Jacalin, AIL). (4) T (Gal beta 1-->3GalNAc), the mucin type sugar sequences on the human erythrocyte membrane(T alpha), T antigen or the disaccharides at the terminal nonreducing end of gangliosides (T beta)--Peanut (PNA), Bauhinia purpurea alba (BPL), Maclura pomifera (MPL), Sophora japonica (SJL), Artocarpus lakoocha (Artocarpin) lectins and Abrus precatorius agglutinin (APA).(5) I and II (Gal beta 1-->3(4)GlcNAc)--the disaccharide residue at the nonreducing end of the carbohydrate chains derived from either N- or O-glycosidic linkage--Ricinus communis agglutinin (RCA1), Datura stramonium (TAL, Thorn apple), Erythrina cristagalli (ECL, Coral tree), and Geodia cydonium (GCL). (6) B (Gal alpha 1-->3Gal), human blood group B specific disaccharide- Griffonia(Banderiaea) simplicifolia B4 (GSI-B4). (7) E (Gal alpha 1-->4Gal), receptors for pathogenic E. coli agglutinin, Shiga toxin and Mistletoe toxic lectin-I (ML-I) and abrin-a. PMID- 9343931 TI - Vegetative tissue lectins of peanut (A. hypogaea). AB - Lectin activities in roots, nodules, stems and leaves of 1-6 week old peanut plant (A. hypogaea) were checked by erythrocyte (human and rabbit) agglutination and sugar inhibition assays. Human and rabbit erythrocyte agglutinating activities were specifically inhibited by lactose/cellobiose (SLII) and methyl alpha-mannoside (SLI) respectively. Seeds, embryos and cotyledons agglutinated neuraminidase treated human erythrocytes and that activity was inhibited by T disaccharide. In the roots of field grown plants SLI was the major activity, while nodules showed both activities (SLI and SLII). Specific activities of SLI and SLII were maximal in stem tissue and hypocotyl exhibited minimal levels. Actively growing tissues like newly emerging young leaves and elongating stem contained more SLII activity in comparison to the mature tissues. Immunological test indicated that all the vegetative tissue lectins are serologically related. PMID- 9343932 TI - Identification of amino groups in the carbohydrate binding activity of winged bean acidic agglutinin. AB - Chemical modification studies reveal that the modification of amino groups in WBA II leads to a complete loss in the hemagglutinating and saccharide binding activities. Since WBA II is a dimeric molecule and contains two binding sites, one amino group in each of the binding sites is inferred to be essential for its activity. The presence of amino group which has a potential to form hydrogen bonded interactions with the ligand, substantiates our observation regarding the forces involved in WBA II-receptor and WBA II-simple sugar interactions. PMID- 9343934 TI - Inhibition of bacterial respiration by a low-molecular weight lectin, scyllin, from Scylla serrata crab hemolymph. AB - Interaction of plant and/or invertebrate lectins with mammalian cells and different microorganisms is well known. In the present study, we have demonstrated that scyllin, a low molecular weight (MW 4000) lectin from the edible crab Scylla serrata hemolymph, purified by GalNAc-Sepharon affinity column followed by Mono-Q ion exchanger in FPLC exhibits antimicrobial activity against Bacillus cereus and Escherichia coli by inhibiting endogenous respiration as well as exogenous glucose oxidation. In both the cases oxygen consumption has been measured in an oxygraph. Scyllin has produced 50% inhibition of endogenous respiration at a concentration of 110 micrograms/ml and 125 micrograms/ml in B. cereus and E. coli respectively. It also reduced the exogenous glucose oxidation by 50% at a concentration of 12 micrograms/ml and 80 micrograms/ml respectively in B. cereus and E. coli. From the above study the mechanism of bacterial growth inhibitory property of scyllin is suggested though the other studies such as inhibition of nucleic acid biosynthesis, cell wall biosynthesis etc. to evaluate its total mode of inhibitory action are not yet obtained. PMID- 9343933 TI - O-acetyl sialic acid binding lectin as a probe for detection of subtle change on cell surface induced during acute lymphoblastic leukemia (ALL) and its clinical application. AB - A novel probe, a 9-O-acetylated sialic acid binding lectin, namely achatininH (ATNH) has been used for the detection of changes on the cell surface during acute lymphoblastic leukemia (ALL). ATNH does not agglutinate normal human erythrocytes, however it is capable of agglutinating erythrocytes and peripheral blood mononuclear cells (PBMC) of patients suffering from ALL. The differential expression of a key receptor, 9-O-acetylated sialo glyco conjugate (9-O-AcSG), on PBMC was observed using a simple lymphoproliferative assay (LA). The extent of expression of 9-O-AcSG was used as an index to distinguish ALL patients of different clinical stages and assess the probability of relapse. The amount of ATNH needed for maximum stimulation served as a tool to indirectly measure the extent of expression of 9-O-AcSG on PBMC surface. The acetylated sialo glycoconjugate was expressed at a very high concentration during acute phase of the disease. Subsequently it decreased during treatment persisted during maintenance therapy and reappeared with relapse. PBMC of normal human donors required 80 times more ATNH in comparison to the untreated acute phase ALL patients. No cross reactivity was found in non Hodgkin's lymphoma, chronic myelogenous leukemia and thalassaemia patients. PMID- 9343935 TI - Ganglioside and neurotrophic growth factor interactions in retinal neuronal and glial cells. AB - Ganglioside (GG) and neurotrophic growth factor (GF) interactions in retinal neuronal and glial cells have been very little studied. Rat retinas were mechanically separated into outer (photoreceptor or PR) and inner (other neurons, IR) halves by planar vibratome sectioning and retinal Muller glial (RMG) cells were isolated and cultured according to previously published methods. The distribution on a percent molar basis of individual GG was different between the two halves: PR were dominated by GD3 (48% total GG) and contained only trace amounts (< 4%) of complex species (GT1b, GQ); IR was more typical of mature brain tissue, exhibiting substantial amounts (approximately 25%) of more complex GG. The GG profile of RMG cells was also simple, dominated by GM3 (60%) and GD1a (20%). A single addition to the medium of 500 pM bFGF or EGF for 48 hr to cultured RMG cells led to significant increases in total GG levels of 30-40%. Such treatments by both growth factors induced increases in GM3, whereas longer exposure (96 hr) of confluent RMG to these factors additionally stimulated synthesis of more complex GG. Incubations of RMG with [3H]-glucosamine showed that GG synthesis was 2-fold stimulated by growth factors. We also tested the effect of GM3 on one of the bFGF receptor transduction pathways, namely PI-3 kinase activation. To our knowledge these data constitute the first demonstration of neurotrophic factor stimulation of GG levels in cells of CNS in vitro. Such complex interactions may have particularly important consequences for neural physiopathology. PMID- 9343936 TI - Cloning and expression of SAT-3 involved in SA-Le(x) biosynthesis: inhibition studies with polyclonal antibody against GST-SAT-3 fusion protein. AB - The SAT-3 activity (CMP-NeuAc:Gal beta 1-4GlcNAc beta 1-3 Gal beta 1-4Glc ceramide alpha 2-3 sialytransferase) involved in the biosynthesis of sialy Le(x) has been characterized in human colon carcinoma cells and embryonic chicken brains. Using RT-PCR-based strategy, we have isolated partial cDNA clones of SAT 3 from ECB and Colo-205 mRNAs. Suitable primers from sialylmotif and N-terminal sequence of human placenta SAT-3 (HP-SAT-3) were used. The 800 bp cDNA fragment encoding a region (90%) of alpha 2-3 sialyltransferase (SAT-3) catalytic domain from ECB has been expressed as a glutathione S-transferase (GST) soluble fusion protein (62 kDa) in E. coli and purified over glutathione-agarose affinity matrix. Polyclonal antibody has been produced against affinity-purified catalytic domain of SAT-3 (GST-SAT-3 fusion protein). A concentration-dependent polydonal antibody binding to native SAT-3 has also been demonstrated by measuring the residual SAT-3 activity in the enzyme fractions from Colo-205. The marked inhibition (> 80%) of SAT-3 activity and relatively less inhibition (< 20%) of SAT-4 activity (CMP-NeuAc:GgOse4Cer alpha 2-3sialyl transferase) suggests strongly the existence of two different gene products (SAT-3 and SAT-4) in human colon carcinoma Colo-205 cells and in embryonic chicken brains (ECB). PMID- 9343937 TI - Glycosyl-phosphatidylinositols in protozoa structure, biosynthesis and intracellular localisation. AB - We are investigating the structure and biosynthesis of glycosyl phosphatidylinositols (GPI) in the protozoa Toxoplasma gondii, Plasmodium falciparum, Plasmodium yoelii and Paramecium primaurelia. This comparison of structural and biosynthesis data should lead us to common and individual features of the GPI-biosynthesis and transport in different organisms. PMID- 9343938 TI - Further characterization of negative regulatory element involved in the hormonal regulation of GlcNAc-1-P transferase gene in mouse mammary gland. AB - The gene encoding UDP-GlcNAc:dolichol phosphate N-acetylglucosamine-1-phosphate transferase (GPT), the enzyme that initiates the pathway for the biosynthesis of asparagine-linked glycoproteins, is ubiquitously expressed in eukaryotic cells. However, its expression in the mammary gland is developmentally and hormonally regulated; transcription of the mouse mammary GPT gene is stimulated by the lactogenic hormones, insulin, glucocorticoid, and prolactin. Earlier, we demonstrated that a distal negative regulatory element in mouse GPT (mGPT) promoter plays an important role in developmental and hormonal control of mGPT gene expression in mammary gland (Ma J, Saito H, Oka T and Vijay IK (1996) J Biol Chem, in press). In this report, a tissue distribution of the repressor that binds the negative regulatory element was examined; a comparison of the negative regulatory element to other consensus sequences for known transcription factors is discussed. PMID- 9343940 TI - The minimal cellular genome of mycoplasma. AB - The mycoplasmas are the smallest and simplest self-replicating organisms, being built of a plasma membrane, ribosomes, and a circular double-stranded DNA molecule-the typical prokaryotic genome. The idea of using mycoplasmas as models for defining in molecular terms the entire machinery of a living cell was raised by Morowitz in 1984. The goal has been to prove the dogma of the completeness of molecular biology, that is, that the logic of life is finite, relatively simple and subject to full exploration. The recent complete sequencing of the genome of the human pathogen Mycoplasma genitalium brings us much closer to achieving this goal. The M. genitalium genome is only 580 kb long and contains only 470 predicted coding sequences(genes), as compared with 1727 in Haemophilus influenzae and about 4000 in E. coli. Thus, M. genitalium is apparently the simplest organism capable of independent life with a minimal set of genes. The drastic economization in genetic information must be associated with the parasitic mode of life of the mycoplasmas. Mycoplasmas evolved by reductive evolution from Gram-positive bacteria with low G + C genomes. During evolution the mycoplasmas have lost the cell wall and many biosynthetic systems involved in synthesis of macromolecule building blocks provided by their host. Thus, the M. genitalium genome carries only one gene involved in amino acid biosynthesis, and very few genes for vitamin and nucleic acid precursors; the lack of genes involved in fatty acid biosynthesis, leads to dependence on exogenous fatty acids, enabling the introduction of controlled variations in membrane acyl chains and the use of mycoplasmas as models in studying membrane fluidity. Moreover, the dependence of mycoplasmas on exogenous cholesterol for growth was exploited to show the role of cholesterol as a buffer of membrane fluidity. The mycoplasma genome carries the minimal set of energy metabolism genes, being content with a restricted supply of ATP needed for their parasitic mode of life. Being limited by a single permeability barrier enabled the saving of a considerable number of transport system genes. Nevertheless, these minimal organisms were shown to carry all the essential genes needed for DNA replication, transcription and translation, but even here gene saving is expressed in a minimal number of rRNA and tRNA genes. A genomic price had been paid to maintain parasitism, so that a significant number of mycoplasmal genes is devoted to adhesins, attachment organelles and variable membrane surface antigens directed towards evasion of the host immune system. PMID- 9343939 TI - Upregulation of the tracheobronchial mucin gene involves cyclic AMP response elements. AB - The cAMP response element (CRE)-binding transcription factor CREB can mediate induction of gene transcription in response to cAMP. Since the tracheobronchial mucin gene (TBM) 5'-flanking region contains CREs (located between residues -289 and -376) with an octamer-like motif (TGACGTCC), the cAMP responsiveness of the TBM CREs was investigated in human tracheal epithelial cells HBE1. These cells were isolated from non-cystic fibrosis subjects and immortalized with HPV18 genes E6 and E7 (ref. 1). HBE1 cells express a homolog of canine TBM (as demonstrated by TBM expression at the transcription and translation level). Electrophoretic mobility shift assay (EMSA) indicated that CREs provide a binding site for nuclear proteins. Transient transfection analysis [using the chloramphenicol acetyl transferase (CAT) reporter gene] and nuclear run on analysis indicated cAMP induced transcription of the TBM gene. The transcriptional activity of the HBE1 transfected cells containing CRE was selectively modulated by extracellular 8Br-cAMP in a dose-dependent manner; a 6-fold increase in activity was detected when cells were incubated for 12 hr in the presence of 2 microM vs 1 nM 8BrcAMP. Since mucin gene is over-expressed in diseases such as cystic fibrosis (CF) and asthma, the information presented here will help us understand the mechanisms involved in transcriptional regulation of mucin gene expression in disease states. PMID- 9343941 TI - Functional and biosynthetic aspects of sialic acid diversity. AB - Sialic acids comprise a large family of N- and O-substituted neuraminic acid derivatives as components of glycoconjugates. N-Glycolylneuraminic acid is formed from N-acetylneuraminic acid by the action of the CMP-N-acetylneuraminic acid hydroxylase studied in various animals. O-Methylated sialic acids originate from the action of S-adenosylmethionine-8-O-methyltransferase studied in starfish. Sialic acids are O-acetylated at diverse positions by the action of acetyl-CoA-4 O- and -7-O-acetyltransferases found in various animals and, leading to the O acetylation of sialic acid glycerol side chain, also in man. Some properties of these enzymes are described and biological implications discussed. PMID- 9343942 TI - Ceramide glycanase from rat mammary tissues: inhibition by PPMP(D-/L-) and its probable role in signal transduction. AB - A ceramide glycanase (CGase activity has been characterized from lactating rat mammary tissue which cleaves the glycosidic bond between sphingosine and the glucose chain of a glycosphingolipid (GSL) thus liberating the intact oligosaccharide chain from a GSL. The majority (65%) of the hydrolase activity was detected in the supernatant fraction when the rat mammary tissue homogenate was centrifuged at 100,000 x g. Attempts to purify the enzyme indicated that the CGase protein is of hydrophobic nature as it binds to hydrophobic columns. The enzyme has been partially purified using hydrophobic columns in tandem. The partially purified protein was found to be immunoreactive to the antibody raised against the purified clam CGase. The immunostained band corresponded to a 64 kDa protein as also found with the clam enzyme. This immuno cross-reactivity indicated probable structural similarities between CGase proteins isolated from widely separated species in the evolutionary tree. The rat CGase was found to have a specific detergent requirement for optimal activity, and the pH optimum was found to be between 5 and 6. The enzyme activity is partially heat stable. It is not a divalent cation requiring enzyme; however, the activity is totally inhibited in the presence of mercury, indicative of a sulfhydryl group in the active site of the enzyme. The rat mammary CGase activity is inhibited in the presence of both D- and L-PPMP (1-phenyl-2-hexadecanoylamino-3-morpholino-1 propanol. HCl), homologs of PDMP (1-phenyl-2-decanoylamino-3-morpholino-1 propanol. HCl), a well-known inhibitor of GlcT-1 (Ceramide: UDP-Glc Glucosyltransferase), an enzyme in the glycolipid synthetic pathway. The inhibition seems to be of a competitive nature and the same type of inhibition is also observed with clam CGase. The CGase activity was found to be highest in lactating tissue compared to the activity found in either pregnant or post lactating rat mammary tissues. Tissue survey indicated the presence of high levels of CGase in lactating rat liver, uterus, and ovary; moderate activity was detected in kidney and spleen. Both virgin and male rat mammary tissue also indicated a basic level of CGase activity. However, newborn spleen and mammary tissue showed a comparable level of activity to that found in lactating rat tissues. This report is mainly concerned with the characterization of CGase activity from a mammalian source and its importance in cellular processes. PMID- 9343943 TI - Use of isotopically radiolabelled GM3 ganglioside to study metabolic alterations in Salla disease. AB - We report the preparation of radioactive GM3 ganglioside and its use in the study of sialic acid storage disorders. For the first time GM3 was isotopically radiolabeled in three positions of the molecule: at the sialic acid acetyl group, [3H-Neu5Ac]GM3, at the C1 of the fatty acid moiety, [14C-Stearoyl]GM3, and at C3 of sphingosine, [3H-Sph]GM3. The radioactive GM3 administered to cultured human fibroblasts from a patient suffering from Salla disease was taken up by the cells and metabolized. An analysis of the distribution of radioactivity within the ganglioside metabolic derivatives showed an accumulation of free sialic acid and ceramide in the pathological cells. PMID- 9343944 TI - Sialylmotifs of sialyltransferases. AB - The sialyl moiety of sialylated glycoconjugates expressed on the cell surface are increasingly recognized as the key determinants of various biological recognition events. The transfer of sialic acid to these glycoconjugates are catalyzed by sialyltransferases, a group of 15 or more Golgi enzymes. Cloning of three sialyltransferases from this laboratory, indicated for the first time, that these enzymes are type II membrane proteins and share the topological features common to other glycosyltransferases. However, unlike the other members of the glycosyltransferase family, these enzymes showed the presence of two conserve protein domains, termed 'sialylmotifs'. This unique feature was subsequently found to be present in all the sialyltransferases cloned to-date. The larger 'L sialylmotif' consisting of 48-49 amino acids is present in the middle of the luminal catalytic domain and has, eight invariant residues, while the 'S sialylmotif' present closer to the C-terminal end of the enzyme has two invariants among a stretch of 23 amino acids. The other not-so-invariant amino acids are also conserved and their replacement is limited. The functional role of these two sialylmotifs were investigated by single-point site-directed mutagenesis using Gal beta 1, 4GlcNAc alpha 2,6-sialyltransferase (ST6Gal I) as a model. Detailed kinetic analysis of the mutants indicated that the 'L sialylmotif' contributes to the binding of the common donor substrate CMP-NeuAc, while the 'S-sialylmotif' contributes to the binding of both the donor and acceptor substrates. PMID- 9343945 TI - Developmental and topographic expression of neutral glycosphingolipids in vertebrate brain. AB - We have examined neutral glycosphingolipid (Ngsl) expression in embryonic (E), post-natal (P) and adult rodent brain employing digoxigenin immunostaining (DIG IS) and anti-Ngsl antisera (both monospecific polyclonal and monoclonal) directed toward specific carbohydrates. Several previously unknown long-chain (-CHO = or > 4) Ngsls have been identified. Four Ngsls have been purified and characterized as GgOse4Cer or GA1, Galactosyl beta 1-3globoside, Fuc alpha 1-3nLcOse4Cer or Lewis X (Le(x)) and a novel GalNAc beta 1-4GA1. A few transient bands appear at different developmental ages. Several fast migrating cerebrosides have also been identified during the early phase of active myelination and tentatively characterized as derivatives of galactosylceramide. Immunohistochemical localization of GA1, Le(x) and GalNAc-GA1 in adult rodent brain shows unique and specific cellular topographies of these carbohydrate antigens. PMID- 9343946 TI - Glycosphingolipid expression in solid tumours and transformed cell lines. AB - Glycosphingolipids are assumed to play a crucial role in cell-cell and cell substrate interactions, including cell adhesion, proliferation, differentiation and apoptosis. Furthermore, cell surface glycolipid profile changes in the so called "social disorders", such as malignant transformation. To better investigate these modifications, the ganglioside composition in different solid tumours and in two transformed cell lines was analyzed. In some of these models we also tried to correlate the pattern of gangliosides to the key enzymes involved in their metabolism. The results we obtained can be summarized as follows:(1), meningiomas with or without chromosome 22 deletion: predominance of ganglioside GD3 in the former and of ganglioside GM3 in the latter. Correlation between GM3/GD3 ratio and SAT-2 activity; (2), mammary carcinomas developed in MMTV/c-neu transgenic mice: accumulation of GM3-derived species. The different ganglioside distribution seems to correlate with the tumour size; (3), Sarcoma Galliera-strain cells SGS/3A and normal syngenic murine fibroblasts FG: transformed cells exhibit a lower activity of sialyltransferases (SAT-1, SAT-2, SAT-4) compared to normal fibroblasts, suggesting a possible correlation with the ganglioside pattern. The neuraminidase activity seems to correlate to the glycoprotein sialic acid content; (4), 3T3 normal murine fibroblasts and SVT2 transformed cells: GM3 is absent in 3T3, while it accounts for the main ganglioside species in SVT2. On the contrary, GM2 present in a large amount in normal fibroblasts, is practically absent in transformed cells. No correlation has been observed between ganglioside profile and glycosyltransferase activities so far examined. PMID- 9343947 TI - Role of phosphorylation and dephosphorylation in the functions of mannose-6 phosphate/insulin like growth factor II receptor. PMID- 9343948 TI - Role of envelope glycoproteins of bovine respiratory syncytial virus in cell fusion. AB - To investigate the requirements for bovine respiratory syncytial virus (BRSV) cell fusion, the fusion (F), the attachment (G) and the small hydrophobic (SH) glycoproteins were expressed individually or coexpressed, using the vaccinia virus-T7 polymerase transient expression system. The contribution of individual glycoproteins in cell fusion was studied by a reporter gene activation assay. Activation of a reporter gene, beta-galactosidase, was assessed by colorimetric assay of detergent cell lysates or by in situ staining. Quantitative measurements indicated much higher sensitivity compared with analysis of syncytium formation. Our results showed that expression of any individual BRSV envelope gene or coexpression of F + G genes, did not induce significant cell fusion; however, coexpression of F + G + SH genes induced extensive cell fusion. To examine the role of N-linked glycosylation, each of the four potential glycosylation sites were individually removed by mutagenesis. The fusogenic activities of these F glycosylation mutants was examined using the reporter gene activation assay. Our results showed removal of individual carbohydrate chains on F2 subunit had no significant deleterious effects on cell fusion. PMID- 9343949 TI - Cytotoxicity of anthrax lethal factor microinjected into macrophage cells through Sendai virus envelopes. AB - Lethal toxin (LT) secreted by Bacillus anthracis consists of two proteins, protective antigen (PA) and lethal factor (LF). LT causes lysis of macrophages and derived cell lines at low concentrations. PA binds to the cell surface receptors and mediates translocation of LF into cytosol of mammalian cells. Internalization of LF into cytosol by osmotic lysis of pinocytic vesicles requires high concentration of LF for cell lysis. To examine the possible cell lysis by LF at low concentration, we introduced LF directly into cytosol of J774A.1 cells through reconstituted Sendai virus envelopes. The introduction of LF lysed J774A.1 cells in a concentration dependent manner. Internalization of PA alone through virosome had no toxic effect on J774A.1 cells. In the process of cytotoxicity LF was not cleaved by cellular proteases. Unlike many protein toxins, golgi was not involved in the expression of lethal toxin activity. These results indicate that LF is the toxic component of anthrax lethal toxin and prior proteolytic processing or trafficking through golgi is not required for its activity. PMID- 9343950 TI - Regulation of DNA replication initiation in mammalian lymphocyte systems. AB - The control of DNA replication is central to the control of cell proliferation, and defects in S phase regulation have been implicated in senescence and neoplasia. To examine the regulation of DNA replication in lymphocytes, an in vitro system was developed in which lymphocyte derived proteins could regulate the initiation of DNA replication in isolated quiescent nuclei. Cytosolic extracts from mitogen or IL-2 activated lymphocytes as well as lymphoblastoid cell lines produce a factor (Activator of DNA replication; ADR) that can induce DNA synthesis in isolated quiescent nuclei, and DNA synthesis in this system is consistent with DNA replication and not repair. ADR activity is tightly associated with a protease activity and is not detectable in resting cells, but can be induced by a mechanism dependent on serine/threonine and tyrosine phosphorylation. Quiescent cells contain an ADR inhibitor which blocks DNA synthesis in isolated normal nuclei but not in nuclei from transformed cells, a potential factor in the uncontrolled proliferation of neoplastic cells. The control of cellular DNA replication is dependent on the interaction of origin sequences with specific replicative and regulatory proteins. However, mammalian origins of DNA replication are not well defined. Plasmids containing a replication origin within the human rRNA gene can act as replicative templates in our cell-free replication system, thus allowing a detailed molecular dissection of replication initiation in a completely human experimental system. PMID- 9343951 TI - Effect of cytokines on tumour cell-endothelial interactions. AB - The adherence of tumour cells to microvascular endothelium is believed to be a necessary step in their migration to sites of metastasis. It has been proposed that this process occurs when cell surface molecules on tumour cells bind to complementary sites on endothelial cells. The expression of these endothelial derived cell adhesion molecules appears to be modulated by cytokines, a broad class of protein mediators which play important roles in immune and inflammatory reactions. It has been found by ourselves and others that exposure of endothelium to some cytokines augments the adhesion of inflammatory cells as well as tumour cells in in vitro assays. We used a murine model consisting of P815 mastocytoma cells and microvascular endothelium and found that pretreatment of endothelial monolayers with TNF-alpha, IL-1, LPS or PMA augmented the number of tumour cells that attach in a dose-dependent fashion. FACS analysis showed that the change in binding was due to an increase in the expression of VCAM-1 on the surface of the endothelial cell. Methylxanthines (caffeine and theophylline) as well as "classical" calcium-mobilizing agents (ionomycin and thapsigargin) inhibited the expression of VCAM-1 in MME. We also studied the possible mechanisms of TNF-alpha signal transduction in endothelial cells. We examined the involvement of protein kinases in the TNF-alpha effect. Although we found that inhibitors of PKC could inhibit the TNF-alpha effect, our studies suggest that the "classical" PKC pathway is not completely responsible for signaling since TNF-alpha did not cause translocation of PKC to the cell membrane and its effect could not be completely mimicked by PMA. We also studied the effect of TGF-beta on the binding of tumour cells to endothelium. Exposure of endothelium to TGF-beta led to the inhibition of both basal and TNF-alpha enhanced binding of P815 cells. Inhibitors of G proteins do not abolish TGF-beta action, and PKC and PKA activators elicit an opposite effect. However, TGF-beta-mediated inhibition of both basal binding and TNF-alpha-enhanced P815 binding to endothelium is completely abolished in the presence of the protein phosphatase inhibitor okadaic acid suggesting that TGF beta elicits its effect by stimulating protein phosphatase activity. PMID- 9343952 TI - An investigation of the nature of bladder mucosal glycoconjugates and their role in interstitial cystitis. AB - The long-term objective of this study is to elucidate the role of bladder mucosal glycosaminoglycans and mucin glycoproteins in the development of interstitial cystitis and other bladder diseases. Bladder biopsies and urine samples from patients and healthy controls were analyzed for glycoconjugates by biochemical and immunochemical methods. Due to the limited availability of human bladders for research purposes, detailed analysis of rabbit bladders glycoconjugates were also carried out. Biochemical analysis of rabbit bladders indicate that while the major portion of the glycoconjugates in the urothelium is sialoglycoprotein, low levels of heparan sulfate and chondroitin sulfate are also present. The correlating immunohistochemical data show very weak staining of the rabbit bladder epithelium by antiglycosaminoglycan antibodies. In contrast, the lamina propria and muscle layers stained intensely for chondroitin sulfate and hyaluronic acid. Thus, the quantity of glycosaminoglycans associated with the bladder epithelial layer, particularly as extracellular matrix components on the luminal surface of the bladder, appears insignificant. On the other hand, several lectins and anti-epitectin (a MUC1 sialoglycoprotein) antibodies showed strong staining of the luminal surface of rabbit and normal human bladders. Further, preliminary results with anti-epitectin antibodies reveal a weaker and patchy staining of biopsy specimens from interstitial cystitis patients compared to controls. The urinary levels of glycosaminoglycans and epitectin, in interstitial cystitis patients and healthy controls were determined by chemical or immunoassays. Urinary epitectin, but not glycosaminoglycans, was decreased in interstitial cystitis patients. PMID- 9343953 TI - Alcohol's impact upon glycobiology. AB - This report reviews and illustrates ways in which some of the problems linked to excessive alcohol intake may develop from alcohol-induced alterations of eukaryotic cell surface molecules. Alcohol is the number one drug of abuse in the US, affecting at least 15 million Americans and causing annual losses of more than $80 billion and 100,000 lives. An estimated 20-40% of all persons admitted to general hospitals have alcohol-related problems and are often undiagnosed alcoholics being treated for the consequences of their drinking. Chronic alcohol related cirrhosis of the liver is the ninth leading cause of death in the US, with over 28,000 deaths annually. Alcohol has harmful effects on almost every organ system in the body, producing cardiovascular disorders, liver disease, neuropathological illness and fetal injury. The etiologic mechanisms for these effects of alcohol is a research area of considerable importance to the National Institute for Alcohol Abuse and Alcoholism. PMID- 9343954 TI - An aminopeptidase regulates LPS stimulated interleukin-8 receptor on the surface of human neutrophils. AB - A large number of inflammatory diseases are mediated by interleukin-8, an inflammatory neutrophil chemotactic agent. Since the cytokine acts through a cell surface receptor, detailed knowledge about the regulation of receptor expression is very important. We found that LPS in serum became activated and triggered the expression of IL-8 receptor by more than two folds within 30 min. After that period, the receptor attained normal level within 2 hr of SA-LPS stimulation. EDTA and bestatin could block this downregulation of IL-8 receptor. Intracellular Ca2+ level was increased till 45 min of SA-LPS stimulation and then the level was reduced. Addition of CaCl2 accelerated and depletion of Ca2+ inhibited the downregulation of the IL-8 receptor. The ligand could fully protect the loss of receptor from downregulation. It suggests that during SA-LPS stimulation, increase in intracellular Ca2+ level activates an aminopeptidase which presumably cleaves the N-terminal region of the receptor, critically essential for the function of IL-8. Thus the activated aminopeptidase regulates the functions of IL 8. The study is important for understanding the regulation of IL-8 receptor expression by LPS during bacterial infection. PMID- 9343956 TI - Importance of glycoproteins in human cancer. AB - Usefulness of cell surface glycoprotein components as markers in early detection of cancer and in monitoring progress during treatment has been evaluated. Total sialic acid (TSA), lipid bound sialic acid (LSA) and seromucoid fractions (SF) have been compared in the sera of healthy human volunteers and patients at different stages of diagnosis and treatment of leukemia, cancer of breast, cervix, and oral cavity. The levels of TSA, LSA and SF are found to be increased in cancer and is proportionate with malignancy. Their levels show decline in patients who respond well to treatment and show increase in patients with recurrence of cancer even before any clinical evidence of recurrence is available. Changes have also been noted in the glycoprotein fractions and their ratios. PMID- 9343955 TI - Collagenase-IV in human trophoblast invasion and differentiation. AB - Trophoblast cells are unique with respect to their functions and responsibilities. These cells demonstrate three sequential phenotypes, proliferation and invasion into the endometrium, differentiation to form syncytia and endocrine secretions. Equipped with these properties placental trophoblasts are endowed with a variety of functions, like implantation of the blastocyst to the endometrium, providing nutrition to the developing embryo and also transmitting extraordinary array of signals for the embryonic development. Experimental evidences and logical extrapolation suggest that these functions are precisely controlled by growth factors, cytokines and hormones produced either by the trophoblast themselves or by the utero-placental unit. Any error in this control mechanism has extremely adverse consequences. The cells also synthesize a large number of enzymes, amongst which collagenase type IV secretion is involved in digestion of underlying basement membrane necessary for the process of invasion. Our results implicate the enzyme in the functional differentiation of the trophoblast as well. Inhibitors to this enzyme inhibit trophoblast differentiation as monitored by secretion of hCG and progesterone, the two markers of trophoblastic differentiation. In contrast, BeWo cells, a choriocarcinoma cell line which does not differentiate spontaneously, undergo increased proliferation when challenged with EGF. The results indicate the possibility of invasive and differentiative phenotypes to be coupled. Exact molecular involvements in this coupling process are looked into. PMID- 9343957 TI - The first 50 pancreas transplants in South Carolina. PMID- 9343959 TI - Reflections on the SMA-1 (A.K.A., the medical history) PMID- 9343958 TI - A child with both congenital fiber type disproportion and giant congenital melanocytic nevi with malignant melanoma. PMID- 9343960 TI - Technology versus responsibility: immigrant physicians from the former Soviet Union reflect on Israeli health care. AB - About 13,000 physicians from the former Soviet Union have found themselves in the saturated medical market in Israel as a result of the latest wave of immigration. This paper examines the gap in professional attitudes and practices between Israeli and Soviet MDs and the cognitive mechanisms employed by immigrant physicians in the process of adjustment to the new medical culture. The study draws on 25 semistructured interviews with recent (about three years in Israel) immigrant doctors who were at various stages of obtaining a local medical license. Reflecting on the need to redefine themselves as professionals and to confront negative stereotypes regarding ex-Soviet doctors, many respondents stressed the strong sides of Soviet medical training and work style. In their collective self-portrait, immigrant doctors emphasized devotion to patients, clinical intuition, manual skills, and empathy, while flaws were regarded as superficial and improvable by technical training. Conversely, the alleged flaws of Israeli doctors were perceived by these informants as pertaining to the core of medicine: "Excessive dependence on technology," "lack of responsibility toward patients," and "weak preventive orientation of Israeli colleagues were repeatedly criticized. The paper sheds light on the significant conceptual differences between the Soviet and Western medical traditions and provides a vivid example of the sociocultural construction of medicine. Our findings are also indicative of the interpretative processes and coping strategies that immigrants in general may develop in saturated professional markets. PMID- 9343961 TI - Effects of specialization and client differentiation on the status of nurses: the case of AIDS. AB - This is a study of how change in the organization of work within hospitals affects the disputes over contested professional jurisdictions. We employ the natural experiment in hospital work reorganization motivated by the AIDS epidemic to empirically document the effects of specialization and client differentiation on the increased intra-organizational status of nurses. We demonstrate that specialized AIDS units represent a form of hospital reorganization in which responsibility, authority, and autonomy devolve toward nurses. Measures of organizational outcomes are derived from the aggregated evaluations of the nurses working in 40 units in 20 hospitals. Our analyses show that different organizational forms are mirrored in differences in the presence of features related to the status and autonomy of nurses. Our work provides some new sociological perspectives on nursing and the changing medical division of labor. PMID- 9343962 TI - The forms and mechanisms of stress proliferation: the case of AIDS caregivers. AB - Processes of stress proliferation are explored in a sample of informal caregivers to people with AIDS. Proliferation refers to the tendency for stressors to beget stressors. Two forms of proliferation are explored, each based on the distinction between primary and secondary stressors. Among AIDS caregivers, primary stressors are the hardships rooted in the caregiving role. Secondary stressors result from primary stressors, but arise in roles and activities outside of caregiving. One form of proliferation is the expansion of primary stressors, reflected in an increase in role overload and a growing sense of being a captive of the caregiver role. Expansion is largely driven by the course of AIDS and the elevation of demands it places on the caregiver. The second form of proliferation is the surfacing of secondary stressors in social and leisure life and in the occupational realm. This form arises from the strains imposed by the emerging caregiver role on the other roles and activities of the caregiver. It is proposed that the systematic assessment of proliferated stressors can help illuminate the dynamic connections between stress and health. PMID- 9343963 TI - Role occupancy and minority mental health. AB - Most studies of the mental health consequences of role occupancy do not consider racial/ethnic variation. Using a national sample of adults (N = 13,017), this paper examines the relationship between three role characteristics (role accumulation, role status, and role combinations) and mental health for Blacks, Mexicans, and Puerto Ricans and explores the extent to which these patterns differ from those for non-Hispanic Whites. Blacks and Puerto Ricans do not benefit from role accumulation whereas Mexicans and Whites who report a high number of roles report better psychological health than those who report few roles. All ethnic groups benefit from the spousal role but there is no consistent effect of either employment or parenthood. Membership in organizational groups benefits non-Hispanic Whites only, whereas familial roles (especially having a sibling) are related to improved mental health among all ethnic groups, except Puerto Ricans. In terms of role combinations, the psychological benefits of occupying all three adult social roles is more evident among non-Hispanic Whites and Mexican Americans compared to Blacks and Puerto Ricans. These findings are discussed in the context of their implications for sociological research which assumes that social psychological processes operate in the same manner across racial/ethnic groups. PMID- 9343964 TI - The meanings individuals attach to role identities and their implications for mental health. AB - Although several theoretical and methodological approaches have been developed for assessing the meaning of roles and role-related stressors, individuals' own understandings of the meaning of their role identities have been ignored in stress research. In this paper, I first examine the ways in which meaning has been conceptualized and assessed. I then explore the meanings individuals themselves attach to role identities and their implications for mental health. Qualitative analyses of indepth follow-up interviews with 40 people who had participated in a community panel study of mental health reveal considerable variation in the meanings they attach to spouse, parent, and worker identities. I also find that the meanings people assign to role identities are based on their perceptions of the benefits and costs of role involvement. Moreover, while most meanings are shared by men and women, there are gender differences in some meanings which reflect gender differences in the perceived benefits and costs of role involvement. Finally, quantitative analyses show that some meanings of role identities are associated with symptoms and are involved in gender differences in distress. These and other illustrative findings suggest that stress researchers would find it useful to incorporate the meanings individuals themselves attach to their role identities and devote greater attention to men's and women's perceptions of both the positive and negative aspects of their role involvement. PMID- 9343965 TI - Education and the subjective quality of life. AB - We examine whether education influences subjective quality of life. If it does, what are the mechanisms by which education affects well-being? We propose that education improves well-being because it increases access to nonalienated paid work and economic resources that increase the sense of control over life, as well as access to stable social relationships, especially marriage, that increase social support. We examine the relationship between education and a variety of indicators of subjective quality of life-depression, anxiety, anger, aches and pains, malaise, and dissatisfaction. Using two representative national samples collected in 1990 and 1995, we find that the well educated have lower levels of emotional distress (including depression, anxiety, and anger) and physical distress (including aches and pains and malaise), but they do not have lower levels of dissatisfaction. Education reduces distress largely by way of paid work, nonalienated work, and economic resources, which are associated with high personal control; but the extent to which it reduces distress by way of marriage and social support is much more modest. We contrast distress and dissatisfaction as indicators of the subjective quality of life. PMID- 9343967 TI - Food for profit, food for thought. PMID- 9343966 TI - Distress and perceived health: mechanisms of health decline. AB - Stress is a common experience in modern society, and it can affect both physical and mental health. Recognizing that not all stress is detrimental to health, this research examines the relationship between perceptions of distress and perceived health within a longitudinal framework. Using two waves of a nationally representative panel study, the National Health and Nutrition Examination Survey I (NHANES I), structural equation modeling revealed that distress leads to more negative health perceptions. In addition, perceived health was found to impact distress levels at the following wave suggesting a cycle of decline between distress and perceived health. Finally, perceived health was found to have predictive validity in determining future functional disability even when considering distress. PMID- 9343968 TI - A prudent and practical approach to the treatment of obesity. AB - Although obesity is difficult to treat, the only effective long-term strategy is an emphasis on adoption of a healthy lifestyle, including exercise, and prudent control over eating habits. Such modifications in behavior can be implemented by some patients with the help and encouragement of their physician, whereas other patients need professional, psychological intervention. VLCDs are useful to attain initial weight loss, but are not (nor were ever intended to be) a substitute for the development of a prudent lifestyle. Thus, for patients with major obesity (BMI > 30), the combination of a VLCD with behavior modification offers the best medical approach to obesity. Surgery has a role in the treatment of obesity, but only in patients with morbid obesity that have repeatedly failed other treatments, including treatments involving a serious effort at behavior modification. PMID- 9343969 TI - Obesity in Arkansas. PMID- 9343970 TI - Diagnosis and treatment of adolescents with eating disorders. PMID- 9343971 TI - Weight loss, nutrition & preventive medicine. A guide for patients. PMID- 9343972 TI - Can aspirin be replaced in the treatment of coronary artery disease? PMID- 9343973 TI - Radiological case of the month. Split peroneus brevis tendon. PMID- 9343974 TI - Special issue on aging. PMID- 9343976 TI - Free radical theory of erythrocyte aging. AB - Mature, circulating mammalian erythrocytes have a finite lifespan. The molecular mechanism that determines removal of cells from the circulation remains unknown, but probably involves recognition of senescence antigens by phagocytes, either directly or via an antibody/complement-mediated pathway. It has been proposed that the major senescence antigen in aged erythrocytes is derived from the band 3 protein, the main transmembrane glycoprotein in erythrocytes. Other possible mechanisms for red cell aging include mechanical fatigue, ATP depletion, calcium accumulation, and the generation of reactive oxygen species (ROS). ROS, which damage proteins and initiate lipid peroxidation, can be generated either inside erythrocytes through the hemoglobin oxidation pathway or outside (eg, by stimulated macrophages). The ROS theory of red cell aging has been widely accepted, yet it lacks direct supporting evidence. To test this hypothesis, two critical techniques have been established in this laboratory. First, we determine the lifespan of erythrocytes in vivo using a fluorescent cell labeling technique. Second, transgenic mice have been produced which express high levels of the human antioxidant enzymes, superoxide dismutase and glucose-6-phosphate dehydrogenase, in their erythrocytes. These two techniques will be very useful for the evaluation of the free radical theory of red cell aging. PMID- 9343975 TI - Mutation and oxidative damage of mitochondrial DNA and defective turnover of mitochondria in human aging. AB - Accumulation of somatic mutations in the mitochondrial DNA (mtDNA) is a major contributor to human aging and degenerative diseases. Rapid progress has been made in unraveling the molecular changes associated with aging. MtDNA mutations are likely early molecular events associated with human aging that may be responsible for the age-dependent decline in mitochondrial respiratory functions. Many types of mutations of the mitochondrial genome impair the function of the respiratory and oxidative phosphorylation systems. This not only results in the age-dependent decline in cellular functions, but also increases the generation of reactive oxygen species (ROS) through the respiratory chain. ROS may cause oxidative damage to mtDNA, further impairing cellular functions and thereby increasing the rate of aging. How aging is elicited by the relatively low amount (< 5%) of aging-associated mutated mtDNA in human tissues is poorly understood. Protein degradation may be a key mechanism in the formation of lipofuscin and possibly in cellular aging. The thiol proteases, critical for protein turnover and degradation, are particularly susceptible to free radical damage at their active sites. If the degradative pathway in mitochondria is defective, the mitochondrion-derived degradation intermediates accumulate within secondary lysosomes, leading to the formation of residual bodies. These degradation products may become another "stressor" to tissue cells. We therefore propose that defective mitochondrial turnover is a cause of accumulation of defective mitochondrial constituents and an important contributory factor to human aging. PMID- 9343977 TI - Regulatory mechanisms of replication growth limits in cellular senescence. AB - Normal human diploid fibroblasts cannot divide indefinitely in culture. At the end of their lifespan they withdraw from the cell cycle permanently by a process termed cellular senescence. Recent molecular studies indicate that upregulation of two inhibitors of cyclin-dependent kinases, p16 and p21, is responsible for blocking the G1/S transition in senescent cells. Although the state of senescence resembles terminal differentiation in that both exhibit irreversible growth arrest and resistance to apoptosis, other molecular changes are seen only in senescent cells. This suggests that the signal pathway specific for senescence is present in normal cells. Changes in chromosomes, such as progressive shortening of the telomeres and erosive damage by detrimental by-products in metabolism, may be the signals that trigger senescence, leading to the inactivation of cell cycle progression. On the other hand, it seems that a dominant genetic program is intrinsically preset to ensure a growth limit in the normal cell. This notion is supported by cell fusion and microcell transfer experiments which show that escaping from senescence requires recessive mutations in senescence-specific genes. Identification of these participating genes and clarification of their mode of action will provide the basis for understanding the mechanisms governing the differences between mortality in normal cells and immortality in cancer cells. PMID- 9343978 TI - Glucocorticoids and aging. AB - In this review we analyze the morphologic changes, hypothalamic-pituitary-adrenal (HPA) axis functions, glucocorticoid (GC) receptors, and steroidogenic enzyme activities in both animals and humans during aging. In rodent studies, older animals tend to show: 1) hypertrophy of adrenal zona fasciculata (ZF) cells; 2) neuronal loss in the hypothalamic area; 3) loss of GC receptors in the hippocampus; 4) raised circulating adrenocorticotropic hormone (ACTH) and GC levels, and increased release of corticotropin-releasing hormone from the hypothalamus; 5) reduced suppression of endogenous GC secretion after administration of dexamethasone; 6) decreased attenuation of response to chronic stress; and 7) increased activity of P450scc and 21-hydroxylase. According to the GC cascade hypothesis, stress and GCs facilitate the aging process in rats. Stress induces downregulation of GC receptors in the hippocampus, then impairs GC feedback on stress-induced HPA axis activation. Finally, an increase in the basal level of corticosterone and extended GC secretion following stress occurs. Because activation of the hippocampus decreases HPA axis function, the unrestrained elevation of GC concentration and the reduction in the level of GC receptors in the hippocampus may gradually weaken the feedback mechanisms and halt the response to stress. In humans, there are conflicting reports of HPA axis function during aging, so it is difficult to make a final conclusion regarding the relationship between aging and HPA axis function. PMID- 9343979 TI - Osteoporotic fracture rate, bone mineral density, and bone metabolism in Taiwan. AB - Taiwanese people have spinal bone mineral density (BMD) values similar to those of Caucasians, whereas their hip BMD values are 10% to 15% lower. In 1992, the prevalence of vertebral fractures, diagnosed according to the -3 SD morphometric criteria, was 18% for women and 12% for men older than 65 years in the major cities of Taiwan. Despite this high prevalence of vertebral fractures, the incidence of hip fractures in the elderly of both sexes was only 203 per 100,000 in 1996, which was lower than in Caucasians and similar to that in mainland Chinese. Hip and vertebral fractures are both associated with lower BMD values. The risk factors for low BMD in Taiwan include a lighter body weight and aging in both sexes, and menopause for women. An increased bone turnover rate is associated with a lower BMD in both men and postmenopausal women, although the rate seems to increase in women but decrease in men with aging. In Taipei City, daily calcium intake is relatively low (mean intake +/- SD; 640 +/- 240 mg), but the vitamin D stores seem to be generally adequate for middle-aged and elderly women. There was a significant association between a higher daily calcium intake and a higher BMD/lower bone turnover rate for women in this age group. Vitamin D receptor allelic polymorphism was not an important factor in low BMD and rapid bone turnover. PMID- 9343980 TI - Demographic characteristics and medical aspects of menopausal women in Taiwan. AB - This report describes the medical and demographic characteristics of menopausal women in Taiwan in order to provide information for consideration during future healthcare planning. The medical and demographic data analyzed were taken from officially published materials of the Government of the Republic of China on Taiwan, our own studies, and those of other researchers. In 1994, the average lifespan of men in Taiwan was 71.8 years and that of women was 77.7 years. The age of menarche was 13.6 years and the age of menopause was 49.5 years. Women aged 50 and over accounted for 18.3% of the total female population and 8.9% of the total population in Taiwan. In 1994, 68.9% of women in Taiwan aged 50 and over were married. The most frequently occurring menopausal symptoms in Taiwanese women were lumbago or low backpain (68%), fatigue (59%), impairment of memory (55%), vaginal dryness (50%), and hot flushes and sweating (49%). The spinal bone mineral density of women decreased markedly after the age of 50 years. The prevalence of vertebral fracture in women 65 years and over was 19.8%, which was higher than the 12.5% in men of the same age group. The prevalence of hypertension and coronary heart disease in women 50 years or older was also higher than in men. The most frequent sites of cancer in women in 1992 were the cervix uteri, breast, sigmoid colon and rectum, lungs, liver, stomach, thyroid, ovaries, hemopoietic and reticuloendothelial systems, and skin. There were 14,298 newly reported cases of malignant neoplasms in women in 1992. About 60% of these occurred in women aged 50 years or more. The median age of occurrence of cervix uteri, breast, and ovarian cancers was 48 to 49 years, which is very close to the menopause age. About 30% of menopausal women in Taiwan are currently living without a husband. Although 18.3% of women in Taiwan were at least 50 years old, approximately 60% of all malignant neoplasms in the female population occurred in this group. There is an urgent need for menopausal women in Taiwan to receive psychologic support and comprehensive medical care. PMID- 9343981 TI - Restoration of sexual behavior in aged male rats by intracerebral grafts of fetal preoptic area neurons. AB - A decline in sexual arousal and copulatory activity has been observed in male rats with advancing age. Grafting of fetal hypothalamic tissue into the third ventricle of aged male rats restores sexual behavior. This study investigated the effects of grafting fetal preoptic area (POA) neurons into the POA of aged male rats exhibiting decreased sexual behavior. We grafted suspensions of fetal POA neurons into the POA of 20 aged (19 to 24 months old) male rats that displayed no ejaculation. From 2 weeks after grafting, one or more series of male copulatory activity and sexual motivation tests were performed at intervals of 3 to 4 weeks. Three behavioral tests given 5 days apart were repeated for each animal in each series. Among the 20 aged rats with the fetal grafts, 15 showed improved sexual motivation, and 14 of these had copulatory activity restored to levels comparable with those of young males. Among these 14 rats, eight ejaculated during copulation. Copulatory behavior was restored between 21 and 45 days after grafting and persisted until the end of observation (2-4.5 months). Sexual performance did not improve in control aged male rats grafted with either fetal cerebral cortex neurons into the POA or POA, neurons into the ventromedial hypothalamus. The rats that received grafts of fetal POA neurons into the POA and recovered sexual performance also showed improvement or recovery to levels comparable to those in young males of serum testosterone concentrations, serum luteinizing hormone levels after castration, and the post-castration rise in luteinizing hormone. These results indicate that decreases in copulatory activity, sexual motivation, and some neuroendocrine functions in aged male rats are at least partially due to dysfunction of the POA. PMID- 9343982 TI - Reversibility of the inhibitory effect of intravesical capsaicin on the micturition reflex in rats. AB - The reversibility of the inhibitory effect of capsaicin on the micturition reflex was investigated in adult rats. The experimental group (n = 38) were given 0.5 mL of 1 mmol/L intravesical capsaicin. After 30 minutes the bladder was evacuated, then the rats were allowed to recover for various times from 0 to 24 hours before cystometrography. Control rats (n = 6) were injected with saline. Immediately after capsaicin treatment, the bladders showed detrusor hyperactivity and high intravesical pressure during cystometrography. All capsaicin-treated bladders showed final urinary retention and hematuria developed in 32 of 38. At 6 hours after intravesical instillation of capsaicin, detrusor hyperactivity was reduced and the micturition reflex gradually reappeared. By 12 hours, micturition reflexes were noted in seven of eight bladders with a volume threshold equal to that of the control group. At 24 hours, the volume threshold for the micturition reflex was significantly greater in the capsaicin-treated group than in the control group. The amplitude of detrusor contractions at 6, 12, and 24 hours showed no significant difference from that in the controls. In vitro whole bladder contractility in response to electrical field stimulation, bethanechol, and KCl also showed no significant difference between the control and the experimental groups. The bladder weight increased as the recovery period increased, indicating the presence of neurogenic inflammation. From this study we conclude that capsaicin-induced micturition reflex inhibition in rats is reversible at 12 hours and the volume threshold for eliciting the micturition reflex continues to increase up to 24 hours after capsaicin treatment. These results may provide insight into the clinical application of capsaicin in the treatment of various voiding disorders in humans. PMID- 9343983 TI - Immunoglobulin M antibody response to hepatitis C virus core protein in patients with chronic hepatitis C. AB - We retrospectively assessed the frequency and clinicopathologic and virologic significance of production of immunoglobulin M (IgM) antibody to hepatitis C virus (HCV) core protein in patients with chronic hepatitis C. Sera from 60 patients with chronic hepatitis C were tested for IgM anti-HCVcore (anti-HCc). Twenty of these patients received ribavirin plus interferon-alpha for 24 weeks, and were classified as sustained, transient, or nonresponders on the basis of alanine aminotransferase levels and the presence of HCV RNA at the end of treatment and 24 weeks later. IgM anti-HCc was detected in 21 patients. There was no correlation between the presence of IgM anti-HCc and clinical features such as sex, age, mode of transmission, serum levels of alanine aminotransferase, HCV genotype, serum HCV titer, or histologic findings. Among the patients who received ribavirin plus interferon-alpha, the mean IgM anti-HCc level before therapy was comparable between sustained (n = 10), transient (n = 8), and nonresponders (n = 2). A statistically significant decrease in IgM anti-HCc response during antiviral therapy was observed in the 18 responders who became negative for serum HCV RNA at the end of therapy. These data suggest that IgM anti-HCc is of limited clinical usefulness as a marker of chronic HCV infection. Serial testing for IgM anti-HCc may provide a marker of antiviral response. PMID- 9343984 TI - Serum estradiol level and oocyte number in predicting severe ovarian hyperstimulation syndrome. AB - Ovarian hyperstimulation syndrome (OHSS) is a relatively common and potentially life-threatening complication of ovarian stimulation, the pathogenesis of which remains unclear. To clarify the predictive values of serum estradiol levels and oocyte number in severe OHSS, and to investigate the impact of high serum estradiol levels on pregnancy outcome, we retrospectively analyzed clinical data from 431 cycles of ovarian stimulation for assisted reproduction performed from 1993 through 1995. Receiver operating characteristic plots were used to estimate the predictive power of the measured variables. The overall frequency of severe OHSS was 5.5%. Using a serum estradiol level of 3,600 pg/mL as the minimum cut off value, the sensitivity was 58%, with a specificity of 92%, a positive predictive value of 29%, and a negative predictive value of 97%. The predictive power was similar when a cut-off point of 20 oocytes retrieved was used. The two criteria together gave a sensitivity of 33%, a specificity of 92%, a positive predictive value of 40%, and a negative predictive value of 98%. One of seven oocyte donors developed severe OHSS. The pregnancy rate was higher in patients with severe OHSS than in patients who did not develop this syndrome (73.9% vs 32.5%) but the pregnancy outcomes were not significantly different. We conclude that elevated estradiol concentrations and oocyte number appear to be helpful in predicting severe OHSS, but neither parameter by itself is predictive. This syndrome is rare in the absence of luteal hCG support, either exogenous or pregnancy-derived; when it occurs, there are usually extremely high preovulatory estradiol concentrations and numerous oocytes retrieved. High serum estradiol levels are unlikely to have adverse effects on pregnancy outcome in patients with severe OHSS. PMID- 9343985 TI - Relapse of acute leukemia in childhood presenting as proptosis due to an orbital mass. AB - Extramedullary relapse of acute leukemia in sites outside the central nervous system or testes is rare. We report two children who developed proptosis due to an orbital mass as the initial presentation of leukemic relapse. The first patient was an 11-year-old girl with acute myelogenous leukemia who had undergone bone marrow transplantation 4 years previously. She had been in remission until 2 months prior to the present admission, when she developed isolated relapse of disease in the left orbit. The second patient, an 11-year-old girl with acute lymphoblastic leukemia, had been off chemotherapy for 2 years and in remission for 5 years. She suffered from concurrent orbital and central nervous system relapse. Both patients were treated with systemic chemotherapy and orbital radiotherapy. More than 1 year after the relapse, the proptosis was completely cured in both patients. PMID- 9343986 TI - Endoscopic third ventriculostomy in the management of non-communicating hydrocephalus. AB - Extracranial diversion of cerebrospinal fluid in hydrocephalus using the currently available shunting system is often unsatisfactory. We have successfully treated four patients with non-communicating hydrocephalus, with no mortality or morbidity, using a flexible endoscope to perform third ventriculostomy. These patients remained shunt independent after treatment and have obtained long-term (minimum 42 months) stabilization. Flexible endoneurosurgical management is simple and safe, and allows in situ observation and performance of biopsies. Endoscopic third ventriculostomy is now our treatment of choice for non communicating hydrocephalus. PMID- 9343987 TI - Influenza. Antigenic analysis of recent influenza virus isolates and influenza activity in the southern hemisphere. PMID- 9343988 TI - Leprosy situation in the world in 1997. PMID- 9343989 TI - Localization of manganese superoxide dismutase in the cerebral cortex and hippocampus of Alzheimer-type senile dementia. AB - Manganese-superoxide dismutase (Mn-SOD) was localized in the cerebral cortex and hippocampus of patients with Alzheimer-type senile dementia (ATD) by immunocytochemistry and the relationship of Mn-SOD with two major pathological features of ATD, i.e., senile plaques and neurofibrillar tangles (NFTs), was examined. Many astrocytes in senile plaques exhibited strong immunoreactivity for both Mn-SOD and glial fibrillar acidic protein (GFAP) on serial section analysis. This suggests that Mn-SOD scavenger system is associated with the formation of senile plaques. On the other hand, Mn-SOD immunoreactivity was not significant in NFT-loaded neurons. PMID- 9343990 TI - Non-operative management of idiopathic ovarian hemorrhage with massive intraabdominal hemorrhage. AB - Although most previous reports recommended operative treatments for ovarian hemorrhages, management of idiopathic ovarian hemorrhage with massive intraabdominal hemorrhage has not been established yet. In order to evaluate effectiveness of non-operative management for idiopathic ovarian hemorrhage, 3 patients with idiopathic ovarian hemorrhage and massive intraabdominal hemorrhage (700-1,400 ml) were managed conservatively and monitored by ultrasonography. Non operative management had successful results in two of the three patients and the other case showed a spontaneous hemostasis of ovarian bleeding at operation. As the first therapy for idiopathic ovarian hemorrhage without any severe complications, non-operative management should be chosen to avoid pelvic adhesions. 700-1,000 ml of intraabdominal hemorrhage were found to be naturally absorbed in a week by ultrasonography. This is the first to report that natural absorption of massive intraabdominal hemorrhage associated with idiopathic ovarian hemorrhage was captured in details by ultrasonography. PMID- 9343992 TI - Localization of the cortical motor area by functional magnetic resonance imaging with gradient echo and echo-planar methods, using clinical 1.5 Tesla MR imaging systems. AB - Functional magnetic resonance imaging (MRI) with gradient echo and echo-planar sequences was applied to healthy volunteers and neurological patients to evaluate the feasibility of detecting and localizing the motor cortex. Time course of the change in signal intensity by an alternate repetition of motor task (squeezing hand) and rest periods was also examined. The motor cortex was localized as the area of signal increase in 88.9% of 45 healthy volunteers by gradient echo method, which mainly reflected the cortical vein, and 83.3% of 30 healthy volunteers by echo-planar method, which mainly reflected the cerebral gyrus. Among 21 volunteers who participated in the both studies, success rate in the localization for the motor cortex was 90.5% (21 volunteers) by gradient echo method and 81% (17 volunteers) by echo-planar method. It was also shown from the time course of the change in signal intensity that signal increase in the most significantly activated area generally corresponded with the periods of the motor task, and the latency between the onset of signal increase and the onset of motor task was usually about 4 seconds. In four of 6 patients with brain tumor, the motor cortex was localized, although activated areas were displaced or distorted. The results indicate that fMRI, either with gradient echo or echo-planar sequence, is a useful method for localizing the primary motor area activated during the motor task and clinically available for noninvasive evaluation of the anatomical relation between brain tumors and the motor area before surgical therapy. PMID- 9343991 TI - Concentrations of bile and serum endotoxin and serum cytokines after biliary drainage for acute cholangitis. AB - Endotoxin contributes to cholangitis. We measured concentrations of bile and serum endotoxin and serum cytokines after biliary drainage for obstructive jaundice with or without acute cholangitis. Patients who underwent percutaneous transhepatic cholangiodrainage (PTCD) in 1995 were classified as having acute cholangitis (group A; n = 11), having a history of acute cholangitis (group B; n = 5), or not having a history of acute cholangitis (group C; n = 13). Bile endotoxin was positive (above the cut-off value) in all patients in groups A and B, and in five patients in group C. The mean concentration of bile endotoxin was significantly higher in groups A and B than in group C. After PTCD, the bile endotoxin level decreased more slowly in group A than in the other groups. Before PTCD, the mean serum levels of IL-1 receptor antagonist (IL-1ra), IL-6, and IL-8 were higher in group A than the other groups. The serum levels of IL-1ra and IL-6 before PTCD were significantly higher when the acute cholangitis was more severe. The mean serum levels of cytokines increased just after PTCD and then decreased. In group A, the serum level of IL-6 at 5 h after PTCD was significantly correlated to the endotoxin level in bile at this time. Increases in cytokines may participate in the pathophysiological changes of acute cholangitis. Biliary drainage for acute cholangitis causes improvement by decreasing the bile endotoxin level addition to decreasing bile-duct pressure, thereby preventing excess production of inflammatory cytokines. PMID- 9343994 TI - In vitro expansion of mature neutrophils from isolated peripheral blood stem cells. AB - Hematopoietic stem cells, CD34 positive cells, were isolated from the peripheral blood of three patients with malignant lymphoma, and were cultivated in suspension for 14 days in the presence of cytokines, including granulocyte colony stimulating factor, granulocyte-macrophage colony-stimulating factor, interleukin 3 and stem cell factor. The stimulation of cell proliferation and differentiation into mature neutrophils was most effective when all these cytokines were used in combination. Mature neutrophils differentiated in vitro gained the ability to ingest latex spheres and to produce hydrogen peroxide in response to phorbol myristate acetate. These findings raise the possibility that the prolonged neutropenia following high dose chemotherapy could be ameliorated by the transfusion of autologous neutrophils expanded and differentiated in vitro. PMID- 9343993 TI - Discrimination of iron deficiency anemia from other anemia by multiple discriminant analysis of routine blood count and red cell distribution width. AB - A computer aided diagnostic program, MDA-3 (Multiple Discriminant Analysis) was designed for clinical use. MDA-3 employs CBC data to analyze using a technique of two-group linear discriminant analysis (a type of multivariate analysis) of anemia and polycythemia. MDA-3 is given discriminant knowledge obtained from a database of 7 CBC items and red cell distribution width (RDW) from 14 groups. We collected 851 CBC data of hematologic abnormalities, designed MDA-3 for IDA screening and evaluated the program's efficacy. The number of cases discriminated as IDA group in the first and the second rank by MDA-3 is up to 80.7%. If false negative cases whose degree of probability is similar to the IDA group are considered as the IDA suspicion group, the diagnostic rate becomes 84.9%. This result demonstrates that MDA-3 is useful for screening of IDA. PMID- 9343995 TI - Reticulocyte maturity index reflects erythropoietin effects in hemodialysis patients. AB - Flow cytometric reticulocyte analysis is widely utilized because of its sensitivity, precision, and high throughput of analysis as compared to manual reticulocyte counting. With this automated method reticulocytes can be classified into three groups according to the fluorescence intensity that reflects maturity of reticulocytes. In this study, changes in red cell parameters, reticulocyte percentage, absolute reticulocyte count, and reticulocyte maturity index were evaluated 13 hemodialysis patients with renal anemia after single administration of recombinant human erythropoietin. The reticulocyte count and reticulocyte maturity index were elevated significantly (P < .01) 2 days after the single bolus of erythropoietin, and they reached the maximum value vn the fourth day, but other red cell indices could not detect the effects of erythropoietin during this early period. These results suggested that the reticulocyte maturity index could be an early indicator of the effect of erythropoietin in hemodialysis patients. PMID- 9343996 TI - Preoperative ultrasound mapping of the saphenous vein: prognostic value on early post operative results, a prospective study. AB - A prospective series of 92 patients had their greater saphenous vein assessed with duplex ultrasound scanning prior to planned infrainguinal bypass procedures. Sixteen (17%) bypass procedures thrombosed within the first week postoperatively. A naturally occurring optimal vein diameter was discovered: 3.5-4.2 mm at mid thigh level and 0-1.5 mm less at mid-calf level. It was significantly correlated with higher one week patency: thrombosis occurred in 11% in veins with this optimal diameter combination and in 19% in all other combination (P < 0.05). These results confirm and expand a retrospective study. PMID- 9343997 TI - Pregnancy and neonatal outcomes in unexplained recurrent/habitual aborters treated by allogenic leukocyte immunization. AB - Allogenic leukocyte immunization is a common treatment of unexplained recurrent abortions, whose immunological side effects have hardly been reported. In this study, pregnancy and neonatal outcomes of unexplained recurrent/habitual aborters, who were treated by their husbands' leukocyte immunization, are reviewed. Seventy-seven patients were treated by their husbands' leukocyte immunization at the Infertility Cline of Osaka City University Hospital from 1985 to 1995. Among the 63 pregnancies after the immunizations, 47 cases succeeded in livebirths. Ten of the 47 cases were delivered by cesarean sections. There found no possible relationship between their operative indications and leukocyte immunizations in the 10 cases. As for pregnancy complications, two patients showed liver dysfunction. However we did not find any specific side effects in the pregnancy complications. There was no specific fetal/neonatal complications to the immunotherapy but a case of neonatal thrombocytopenia caused by maternal anti-HLA IgGs. PMID- 9343998 TI - Favorable response to heparin in a pregnant woman with possible glomerular thrombosis as a complication of systemic lupus erythematosus and anti phospholipid antibody syndrome. AB - A rare case of possible glomerular thrombosis during pregnancy is reported in a patient with active systemic lupus erythematosus and the presence of anti phospholipid antibodies. Acute renal impairment was restored by administering infusions of heparin. Cesarean section was performed due to fetal distress, and resulted the live birth of a healthy infant. PMID- 9343999 TI - A case of membranous glomerulonephritis associated with gastric cancer. AB - We describe a patient with gastric cancer and membranous glomerulonephritis (MGN). The patient, a 61-year-old male, was admitted to our Hospital in May, 1996, because of proteinuria and hyperlipidemia persisting for a year. Laboratory examination filled the criteria of nephrotic syndrome and renal biopsy revealed MGN of stage II. Prednisolone therapy (40 mg/day p.o.) was started, followed by a gradual decrease in proteinuria from 4.5 g/day to 0.1 g/day. Endoscopic examination was performed because of stomach-ache revealed advanced gastric cancer of Borrmann 4. Desiring for a conservative therapy, he was discharged and moved to a hospice. In literature review, MGN is the most frequent lesion among various glomerular diseases associated with malignancy, such as the lung, stomach, and colon, particularly at an elderly ages, and sometimes antedates the detection of malignancy, as in the present case. In several cases with MGN, immune-complexes composed of tumor antigens, such as carcino-embryonic antigen, and antibodies have been reported to deposit in basement membrane of glomeruli, causing MGN. In the renal and gastric cancer tissues of the present case, the presence of three novel tumor-associated antigens, Span-1, Thomsen-Friedenreich antigen (T antigen) and F1 alpha antigen, was examined, using a immuno-peroxidase method. Although none of these three antigens were immuno-stained in the renal tissue, clinical course and literature review suggest that MGN in this patient seems to be associated with gastric cancer, which may have produced MGN-causing tumor antigens other than the three antigens. It should be emphasized that malignancy should be carefully and routinely examined in patients with MGN, particularly at elderly ages. PMID- 9344000 TI - Intraoperative color duplex sonography of basal arteries during aneurysm surgery. AB - This prospective study aimed at (1) characterizing the duplex sonographic appearance of cerebral aneurysms, (2) visualizing their location, and (3) ensuring the complete occlusion of the aneurysm as well as the patency of the basal arteries during aneurysm surgery. During 9 months 30 craniotomies for aneurysm clipping in 29 patients were monitored intraoperatively by B-mode and color-coded duplex sonography. Following craniotomy the aneurysm and the preaneurysmatic and postaneurysmatic arteries were sonographically visualized before and after clipping and removal of the spatulas. Twenty-seven (90%) of 30 aneurysms appeared as a hypoechoic structure. Together with the typical dichromatic picture in the color mode and the characteristic bidirectional flow pattern in the duplex mode, 29 (97%) of 30 aneurysms were identified and localized anatomically correctly. Eighty (99%) of 81 relevant vessels were visualized and measured with the Doppler mode. After clipping, flow was detectable in all major arteries except 3 middle cerebral artery (MCA) branches. In 1, occlusion was confirmed by postoperative angiography. In the other 2, early postoperative computed tomography showed an infarction of the corresponding MCA territories. This study demonstrated the potential of color duplex sonography to visualize and characterize cerebral aneurysms and adjacent basal arteries before and after clipping. It offers a noninvasive intraoperative method to control the patency of basal arteries and complete occlusion of the aneurysm. PMID- 9344001 TI - Degeneration of the pyramidal tracts in patients with amyotrophic lateral sclerosis. A premortem and postmortem magnetic resonance imaging study. AB - To investigate focal hyperintensity in the internal capsule (IC) on magnetic resonance images (MRIs) and its clinical significance, 80 patients with amyotrophic lateral sclerosis (ALS) and 80 sex- and age-matched normal control subjects were studied. On T2-weighted images, hyperintense foci were found in the posterior part of the posterior limb (PL) of the IC in 41 (51%) of 80 control subjects. However, no subject showed increased signal intensities on proton density-weighted images. Hyperintense foci were also observed in the posterior part of the PL of the IC on T2-weighted images in 52 (65%) of 80 ALS patients and on proton density-weighted images in 26 (65%) of 40 ALS patients; the abnormally intense foci were seen at the same anatomical location in the IC as those in the normal control subjects. On postmortem MRI, the abnormally intense foci were found in the posterior part of the PL of the IC in the formalin-fixed brains from 9 ALS patients. Three normal control subjects did not show signal intensity changes on postmortem MRI. On histological examination of 9 ALS brains, distinct myelin pallor and gliosis were found in the posterior third of the PL of the IC. Proton density-weighted images appear to be useful to distinguish neuropathological changes in the corticospinal tract of ALS patients. PMID- 9344003 TI - Microemboli on transcranial Doppler in patients with spontaneous carotid artery dissection. AB - High-intensity transient signals on transcranial Doppler sonography (TCD) are associated with atherosclerotic stenosis of the internal carotid artery. Few data exist regarding the detection of high-intensity transient signals in dissected carotid arteries. In the present study, 6 patients with spontaneous carotid dissection, defined by magnetic resonance techniques and duplex sonography, were examined by TCD. Both middle cerebral arteries were monitored simultaneously for 30 minutes. Four of the patients had ipsilateral cerebral ischemia, while 2 presented with other symptoms. High-intensity transient signals were detected in the middle cerebral artery ipsilateral to the dissection in 3 of the 4 patients with cerebral ischemia and in none of the patients with other presenting symptoms. No microemboli were found contralateral to the dissected arteries. Microemboli can be detected distally from dissected carotid arteries. The present findings support the assumption that embolism is a major cause of stroke in patients with carotid dissection, and suggest that high-intensity transient signals are more common among patients with cerebral ischemia secondary to dissection. PMID- 9344002 TI - Transcranial Doppler in 178 patients before, during, and after carotid endarterectomy. AB - From July 1991 to March 1995, 178 patients who underwent 198 carotid surgical repairs were investigated preoperatively, intraoperatively, and postoperatively by transcranial Doppler sonography (TCD). Preoperative TCD evaluation showed stenosis of the middle cerebral artery (MCA) in 4 patients (2.2%), siphon stenosis in 3 (1.6%), incomplete circle of Willis in 23 (12.9%), a decrease of mean blood flow velocity more than 70% of the basal value during digital common carotid compression in 31 (17.9%), and a critical reduction of vasomotor reactivity (no significant increase of mean blood flow velocity in the MCA during breath-holding test) in 34 (19.1%). Nine patients (5%) had surgery without preoperative angiography. In those patients the indication for surgery was based on color Doppler imaging and TCD investigations. Ninety surgical procedures were carried out under general anesthesia and 188 under locoregional anesthesia. In 37 surgeries (31.7%) a shunt was inserted. The use of a shunt was based on a decrease of mean blood flow velocity in the MCA below 50% of the basal value under general anesthesia or loss of consciousness combined with a decrease of mean blood flow velocity in the MCA higher than 70% of the basal value when locoregional anesthesia was employed. Intraoperative TCD monitoring showed a decrease of mean blood flow velocity in the MCA due to shunt malfunction in (8.3%) of 36 surgeries, turbulence of blood flow during declamping in 79 procedures (39.8%), and microembolic events in 10 patients (5%) that were related to one transient and one permanent neurological deficit. Another permanent deficit occurred in a patient without TCD signs. After surgery, TCD reliably detected an early asymptomatic occlusion of the carotid artery, hyperperfusion syndrome in 12 (6.0%), and an increase of vasomotor reactivity in 10 (29.4%) of 34 surgeries. PMID- 9344004 TI - Power Doppler compared to color-coded duplex sonography in the assessment of the basal cerebral circulation. AB - Power-based transcranial duplex sonography (p-TDS) is a new promising ultrasound technique that generates intravascular color signals from the amplitude of the echo signal. The present investigation was undertaken to determine the advantages and limitations of power Doppler in the assessment of the basal cerebral circulation compared with transcranial color-coded real-time sonography (TCCS) and contrast-enhanced transcranial color-coded real-time sonography (CE-TCCS). Thirty-eight patients without cerebrovascular diseases were examined with p-TDS and TCCS, and in 11 patients CE-TCCS studies were performed. The M1 segment could be identified in 100% by both ultrasound techniques. p-TDS visualized M2 (67/70 vs 46/70, p < 0.0001), A2 (63/70 vs 46/70, p < 0.001), and P2 (67/70 vs 44/70, p < 0.0001) segments significantly more frequently and accurately compared to TCCS. The posterior communicating artery (25/70) and P3 segments (32/70) were only detectable by p-TDS and not by conventional TCCS. In comparison with CE-TCCS, p TDS had no important advantages in the detection of intracranial vessels. In conclusion, p-TDS and CE-TCCS were superior to TCCS with regard to identification of the basal arterial circulation. Both methods permit noninvasive and reliable identification of the basal cerebral circulation. PMID- 9344005 TI - Diffusion-weighted magnetic resonance imaging characteristics of hemorrhagic transformation in experimental embolic stroke. AB - Diffusion-weighted magnetic resonance imaging (MRI) can detect ischemia within minutes of onset, but its ability to reliably detect hyperacute cerebral hemorrhage is unknown. The present study characterized diffusion-weighted, T2 weighted, and contrast-enhanced T1-weighted MRI appearances of hemorrhagic transformation within 5 hours of onset in experimental embolic stroke. Apparent diffusion coefficients and MRI signal characteristics were noted within corresponding regions of hemorrhage observed on gross pathology. Apparent diffusion coefficients were significantly increased within hemorrhagic lesions, but were still within the expected range for bland ischemia. The appearance of the hemorrhagic lesions on diffusion-weighted MRI was also very heterogeneous and not very useful for clinical screening. Other MRI modalities should be investigated, but computed tomography remains the only widely available clinical method of reliably detecting cerebral hemorrhage. PMID- 9344007 TI - Algorithm on the clinical evaluation of epilepsy. AB - Epilepsy is an important public health problem because of its prevalence in the community and the economic disadvantage associated with its chronic morbidity. Uncontrolled seizures are potentially life-threatening and have adverse psychosocial consequences. Surgery is an effective but underutilized therapy for some patients with medically refractory seizures. However, this form of treatment demands precise localization of the epileptogenic zone. In recent years there have been major advances in the development of various imaging techniques for seizure localization. The best combination of diagnostic testing with regard to cost-benefit has been debated. A rational strategy for the deployment of these techniques is discussed. PMID- 9344006 TI - Balloon angioplasty of intracranial arteries for stroke prevention. AB - Stroke from surgically inaccessible intracranial atherostenosis remains a formidable clinical challenge. While antithrombotic or antiplatelet therapy may prevent distal embolism, there is no effective program for plaque stabilization preventing progression of atherosclerotic stenosis. In patients with isolated circulations (single vertebral with absent posterior communicating arteries, single carotid with contralateral internal carotid artery occlusion, or single carotid with an absent anterior communicating artery), occlusion of the stenotic vessel may produce a low flow-mediated stroke. Fifteen patients with atherosclerotic intracranial stenoses were treated by balloon angioplasty after medical therapy with warfarin failed. Treated territories included the distal internal carotid, proximal middle cerebral, distal vertebral, and basilar arteries. Dilation was successful in all vessels, with residual stenoses averaging less than 30%. Two complications included one paramedian pontine stroke and a single vessel rupture that proved fatal. There was no recurrence of transient ischemic attacks and no restenosis at the angioplasty site over a follow-up period of more than 24 months. In this small series, balloon angioplasty of intracranial vessels provided a therapeutic option for secondary stroke prevention in highly selected patients. Further studies will be necessary to establish the efficacy and safety of endovascular treatment in larger series. PMID- 9344008 TI - Spinal leptomeningeal hemangioblastomatosis in von Hippel-Lindau disease: magnetic resonance and pathological findings. AB - A 55-year-old man with von Hippel-Lindau disease presented with quadriparesis. Multiple enhancing cervical and thoracic spinal masses were seen on magnetic resonance imaging (MRI). A rim of diffuse, nodular enhancement linking all of the discrete masses was apparent on the surface of the cervical and thoracic regions of the cord. Surgical exploration revealed multiple extramedullary-intradural and intramedullary masses, extending to and infiltrating the cord; the leptomeninges contained numerous small tumor seeds at several levels. The excised spinal masses were diagnosed as capillary hemangioblastomas, which infiltrated the pia mater. Diffuse, intense, spinal leptomeningeal enhancement on MRI associated with multiple hemangioblastomas has not been previously reported and may be referred to as spinal "leptomeningeal hemangioblastomatosis." PMID- 9344009 TI - Magnetic resonance cisternography in the diagnosis of delayed iatrogenic cerebrospinal fistula: a case report. AB - A 52-year-old woman presented with a clinical picture consistent with bacterial meningitis 3 years after functional endoscopic sinus surgery. Diagnosis of a cerebrospinal fluid (CSF) fistula was made clinically, and the site of the fistula was confirmed using magnetic resonance cisternography. The utilization of this technique in the diagnosis of CSF disorders is gaining popularity. Its usefulness in the context of other imaging modalities is discussed. PMID- 9344010 TI - Extensive reversible brain magnetic resonance lesions in a patient with HELLP syndrome. AB - A severe form of toxemia of pregnancy with microangiopathic hemolytic anemia, elevated liver enzymes, and low platelets has been called the HELLP syndrome. A patient with the HELLP syndrome developed a severe, reversible encephalopathy. Brain computed tomography and magnetic resonance imaging showed abnormalities consistent with edema limited to the posterior circulation territory. The location of the lesions and their occurrence in the HELLP syndrome support suggestions that the vulnerability of posterior structures in eclamptic encephalopathy is due to a vascular susceptibility of the posterior circulation and that endothelial cell dysfunction plays an important role in the pathogenesis of eclamptic encephalopathy. PMID- 9344011 TI - Acute bilateral infarcts of the posterior inferior cerebellar artery. AB - Acute bilateral infarcts in the territory of the posterior inferior cerebellum artery are rare and poorly documented in the literature. Thus, this report describes the clinical course and outcome in 3 patients. Although one was associated with coronary artery bypass surgery, the etiology was not known. Despite large territorial infarcts, the patients recovered to ambulation with minimal assistance. PMID- 9344013 TI - Do right by the disabled. PMID- 9344014 TI - Six myths about physician executives. PMID- 9344012 TI - Ultrasonic bruits in the circle of Willis due to a large nonfunctioning pituitary adenoma. AB - In a man with a histologically verified non-functioning pituitary adenoma with suprasellar extension, Doppler signals resembled those associated with bruits (ultrasonic bruits). These signals were detected in the anterior circulation of Willis both preoperatively and intraoperatively. The large tumor was resected subtotally via a right orbitozygomatic approach. The use of microvascular sonography for intraoperative monitoring can provide information on the potential cerebrovascular complications of surgery. No previous studies on the presence of ultrasonic bruits associated with pituitary adenomas have been reported. The clinical implications for the surgical treatment of pituitary adenomas are discussed. PMID- 9344015 TI - Consumers. Public distrust. PMID- 9344016 TI - Quality Patrol. Keep it clean. PMID- 9344017 TI - Domestic violence ... people who show up in the ER. PMID- 9344018 TI - Human resources ... shouldn't focus only on the basics of hiring and firing. PMID- 9344019 TI - Technology ... information systems are fast-growing, big-ticket items. PMID- 9344020 TI - Finance ... sharing risk with an insurer. PMID- 9344021 TI - Patient records. Who do they trust? PMID- 9344022 TI - Integration ... competing successfully by "carving out" specialties. PMID- 9344023 TI - The subtext is reform. As the quality commission hashes out its mandate, sticky questions keep coming up. PMID- 9344024 TI - Rough crossings. AB - CEOs change jobs all the time, but only a handful make the jump from not-for profit to for-profit hospital or vice versa. Some thrive in the new setting. Others crash. Personality makes the difference. PMID- 9344025 TI - My brother's gatekeeper. AB - When doctors launch their own managed care ventures, they soon find that cost control means care control. And in policing their own, they must resort to the techniques used by other health plans--gatekeepers and all. PMID- 9344026 TI - PAYOFF@InfoTech. NOW. AB - Why shouldn't your computer system earn its keep? Tracking the return on investment is simply good business discipline. But some insist that traditional cost/benefit analyses don't apply. PMID- 9344027 TI - Doc stocks. AB - Wall street's starry-eyed infatuation with practice management companies is gone, causing a market correction that's deflated stock multiples and slowed acquisitions. But don't write off the romance yet. PMID- 9344028 TI - NovaCare's big bang. AB - The no. 2 rehab provider took a dose of its own medicine--therapy that boosted productivity, slashed costs, and pushed the company into new markets. Now a consensus of analysts sees growth soaring by 20 percent a year. PMID- 9344029 TI - Stuck on a strategy. Interview by Mary Grayson. PMID- 9344030 TI - Pathways to nowhere. AB - Early in the decade, health care discovered a powerful patient care management tool: the clinical path. Leaders in the field latched onto paths as the solution to cost and quality dilemmas, but they overlooked the importance of the underlying principles of quality management. Beyond Clinical Paths, a new book from American Hospital Publishing, focuses on the issues critical to successful quality improvement through clinical paths. The following excerpt by the book's editor, Patrice L. Spath, sets the stage for a new approach to clinical quality. PMID- 9344032 TI - AIDS drugs. Dramatic twist on coverage. PMID- 9344031 TI - Rural managed care. The Maine event for small companies. PMID- 9344033 TI - Patient advocacy. Legal remedies. PMID- 9344034 TI - Hospital pulse ... May 1997. PMID- 9344035 TI - Mission guardian. More a calling than a job. Interview by Chuck Appleby. PMID- 9344036 TI - Quality patrol. A fix for drug errors. PMID- 9344037 TI - Health care consulting. When only a doc will do. PMID- 9344038 TI - Online benefits. A net for new business. PMID- 9344039 TI - Marketing to women. Clinics with a feminine touch. PMID- 9344040 TI - Nitric oxide induced by tumor cells activates tumor cell adhesion to endothelial cells and permeability of the endothelium in vitro. AB - Human fibrosarcoma HT1080 cell surface phenotype analysis revealed the expression of "cluster of differentiation 15" (CD15) antigen and to a lesser extent, of "very late antigen-4" (VLA-4). Expression of "endothelial-leukocyte adhesion molecule-1" (ELAM-1) was negligible on resting human umbilical vascular endothelial cells (HUVECs), but its expression could be induced by HT1080 conditioned medium. HT1080 cell adhesion to HUVECs was partially dependent on CD15/ELAM-1 adhesion molecules. HT1080 cell adhesion to HUVECs induced the enhancement of nitric oxide (NO) production from HUVECs. Exogenous NO and NO from HUVECs enhanced ELAM-1 expression on HUVECs, HT1080 cell adhesion to HUVECs, permeability of the HUVEC monolayer, and HT1080 cell invasion through the HUVEC monolayer. These enhancements were not induced by NO synthase inhibitor, N(G) nitro-L-arginine methyl ester (L-NAME). These results suggest that NO expression induced by tumor cells via the CD15/ELAM-1 adhesion system may contribute to enhancement of tumor cell adhesion to endothelial cells and hyperpermeability of the endothelium, facilitating tumor cell invasion. PMID- 9344041 TI - Activation of protein kinase C-alpha isoform in murine melanoma cells with high metastatic potential. AB - Metastasis is a multistep process in which protein kinase C (PKC) appears to be significantly involved. We analysed the activity and expression of classical (alpha, beta, gamma) and novel PKC epsilon isoforms in B16-F1 and B16-BL6 melanoma cells maintained under different culture conditions in vitro. We used high and low concentrations of tyrosine and phenylalanine in different media (DMEM or RPMI 1640 respectively) that affect the metastatic potential and also the proliferative capacity of the cells. We also tested a weakly metastatic amelanotic B78-H1 melanoma cell line which is unaffected by the different culture conditions. In both B16 melanoma cell lines activation of PKC alpha (without increased expression) occurred under growth conditions permissive of metastasis (DMEM). In contrast, the weakly metastatic amelanotic B78-H1 cell line showed a substantial inactivation of this isoform in the two different culture media, suggesting a specific involvement of PKC alpha in the metastatic process. Moreover, in B16 melanoma cells, novel PKC epsilon was activated under culture conditions which stimulated growth but not metastasis (RPMI 1640). In order to define the relationship between PKC activation and the metastatic process we also determined the release of cathepsin B. No correlation between PKC activity and cathepsin B release in either B16 melanoma cell lines could be demonstrated. PMID- 9344042 TI - Calcitriol enhancement of TPA-induced tumorigenic transformation is mediated through vitamin D receptor-dependent and -independent pathways. AB - We previously showed that 1alpha,25-dihydroxyvitamin D3, calcitriol, enhanced phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced tumorigenic transformation of mouse epidermal JB6 Cl41.5a cells. To determine if calcitriol regulates this enhancement through a nuclear vitamin D receptor (VDR)-dependent or -independent pathway, we used vitamin D analogs which induce biological responses by either of these mechanisms. In JB6 Cl41.5a cells, 1alpha,24 dihydroxy-22-ene-24-cyclopropyl-vitamin D3 (BT), which like calcitriol binds to VDR and regulates transcription, inhibited cell growth, stimulated expression of nonphosphorylated osteopontin (OPN), and enhanced TPA-induced anchorage independent growth (AIG, an in vitro assay which highly correlates with tumorigenicity of these cells). 25-Hydroxy-16-ene-23-yne-vitamin D3 (AT), which stimulates calcium influx but has low affinity for VDR, had moderate effects on cell growth and expression of OPN. However, it enhanced TPA-induced tumorigenic transformation, though to a lesser extent than BT, thus suggesting that a VDR independent mechanism is involved. Since 1alpha-hydroxylase activity was detected in JB6 cells, AT could be converted into 1alpha,25-dihydroxy-16-ene-23-yne vitamin D3 (V), an analog which binds with high affinity to VDR, and could subsequently enhance TPA-induced AIG. To verify whether the VDR-independent pathway is involved in calcitriol enhancement of tumorigenic transformation, two additional VDR-independent analogs, 1alpha,25-dihydroxy-lumisterol3 (JN) and 24R,25-dihydroxyvitamin D3 (AS), were tested. The analog JN, which stimulates calcium transport and cannot be further hydroxylated at 1-carbon position, increased TPA-induced AIG, while AS, which inhibits calcium influx, did not. These studies suggest that a VDR-independent pathway, perhaps stimulation of calcium influx, and a VDR-dependent mechanism, which directly affects transcription, are involved in calcitriol's enhancement of TPA-induced tumorigenic transformation in JB6 Cl41.5a cells. PMID- 9344043 TI - Regulation of desmosomal cell adhesion in human tumour cells by polyunsaturated fatty acids. AB - Desmosomes are key structures in cell-cell adhesion. In this study we examined the effect of n-6 essential fatty acids on the expression of desmoglein (Dsg), desmosomal cadherin and the formation of desmosomes in E-cadherin negative human breast, colon and lung cancer cells and melanoma cells. Electron microscopy revealed that cells cultured with gamma linolenic acid (GLA) showed increased cell-cell adhesion together with an increase in the formation of desmoglein containing desmosomes. Western blotting studies of cellular proteins demonstrated that, following culture with fatty acids, Dsg expression was modified, with the greatest increase seen after GLA treatment. Other fatty acids increased Dsg expression, but to a lesser extent. It is concluded that GLA regulates desmosome mediated cell-cell adhesion in human cancer cells, particularly in cells without E-cadherin. PMID- 9344044 TI - Inhibition by ginsenoside Rg3 of bombesin-enhanced peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane in Wistar rats. AB - The effects of concomitant use of bombesin and ginsenoside Rg3 on the incidence of peritoneal metastasis of intestinal adenocarcinomas induced by azoxymethane were investigated in male inbred Wistar rats. From the start of the experiment, rats were given weekly s.c. injections of azoxymethane (7.4 mg/kg body weight) for 10 weeks and s.c. injection of bombesin (40 microg/kg body weight) every other day, and from week 20, s.c. injections of ginsenoside Rg3 (2.5 or 5.0 mg/kg body weight) every other day until the end of the experiment in week 45. Bombesin significantly increased the incidence of intestinal tumors and cancer metastasis to the peritoneum in week 45. It also significantly increased the labeling index of intestinal cancers. Although administration of a higher dose of ginsenoside Rg3 with bombesin had little or no effect on the enhancement of intestinal carcinogenesis by bombesin, the location, histologic type, depth of involvement, infiltrating growth pattern, labeling and apoptotic indices and tumor vascularity of intestinal cancers, it significantly decreased the incidence of cancer metastasis. These findings indicate that ginsenoside Rg3 inhibits cancer metastasis through activities that do not affect the growth or vascularity of intestinal cancers. PMID- 9344045 TI - Increased matrix metalloproteinase-9 activity in human ovarian cancer cells cultured with conditioned medium from human peritoneal tissue. AB - Ovarian cancer cells disseminate by attachment to the peritoneal mesothelial cell surface of the abdominal cavity. We therefore investigated the influence of conditioned medium (CM) from human peritoneal tissues and mesothelial cells on the secretion of matrix metalloproteinases (MMPs) by ovarian cancer cells. The molecular weights of MMPs stimulating factors derived from human peritoneal tissues and mesothelial cells were estimated using microconcentrators with various cut-off membranes. Human peritoneal tissues were obtained from 12 surgical patients, and mesothelial cells were isolated from three peritoneal specimens. Exposure to CM from peritoneal tissue caused a concentration-dependent increase of the MMP-2 and MMP-9 bands in CM from NOM1 ovarian cancer cells, as shown by zymography. There was a significant difference in the increase of MMP-2 and MMP-9 (2.46-fold and 7.14-fold, respectively, at 0.4 mg/ml protein; P < 0.005). CM from mesothelial cells also significantly increased the secretion of MMP-9 by NOM1 cells. The molecular size of possible MMP-9-stimulating factors secreted by peritoneal tissues and mesothelial cells was above M(r) 100000. Further, CM of peritoneal tissues and mesothelial cells also induced the invasiveness of NOM1 cells. These findings suggest that mesothelial cells may secrete some factors which predominantly induce the MMP-9 production and increase invading cell numbers. PMID- 9344048 TI - Partnerships: challenges and rewards. PMID- 9344047 TI - Spontaneous but not experimental metastatic activities differentiate primary tumor-derived vs metastasis-derived mouse prostate cancer cell lines. AB - We previously developed an in vivo mouse prostate reconstitution (MPR) model of metastatic prostate cancer using p53 'knockout' mouse urogenital sinus tissue for retroviral transduction of ras and myc oncogenes (Thompson et al., Oncogene, 10, 869, 1995). We further demonstrated contrasting responses to transforming growth factor beta-1 (TGF-beta1) in three matched pairs of early passage cell lines derived from primary prostate tumors and lung metastases generated by this model system (Sehgal et al., Cancer Res, 56, 3359, 1996). In this study we tested these cell lines for growth potential in subcutaneous and orthotopic (dorso-lateral prostate) locations and metastatic activities in both spontaneous and experimental assays. Subcutaneous and orthotopic tumors produced by cell lines derived from metastatic lesions tended to grow less rapidly but demonstrated greater spontaneous metastatic potential than the cell lines derived from primary tumors. In contrast all cell lines produced lung colonies in an experimental metastasis assay (tail vein inoculation) with the primary tumor-derived cell lines yielding higher activities in two of three matched pair analyses. The ability of all cell lines to produce lung metastases in the experimental assay, while only the metastasis-derived cell lines retain the ability to initiate and complete the entire metastatic pathway in the spontaneous assay, suggests that intravasation may be the rate-limiting step in metastasis in this model system. PMID- 9344046 TI - Effect of fibroblast growth factor saporin mitotoxins on human bladder cell lines. AB - Mitotoxins targeted via high-affinity growth factor receptors on the cell surface are a potential means of anticancer therapy. We have evaluated the effect of a chemically conjugated (FGF2-SAP) and a fusion protein (rFGF2-SAP) mitotoxin containing FGF-2 and saporin on normal (FHs 738B1) and malignant bladder cell lines (HT1197, TCCSUP, EJ-6, and RT4). The FGF-saporins demonstrated potent cytotoxicity in malignant bladder cell lines with an ID50 range of 0.13-13.6 nM, whereas cells derived from normal fetal bladder (FHs 738B1) were less sensitive to FGF2-saporins (ID50 > 100 nM). Greater than a 100-fold difference in cytotoxicity between FGF-saporins and unconjugated saporin was observed. Assessment of cellular FGF-2 content and secretion showed that FHs 738B1 and TCCSUP contained and secreted significantly more FGF-2 compared to other cell lines tested. (125)I-FGF-2 receptor binding studies showed the presence of high affinity (pM) FGF receptors on all bladder cell lines. Cross-linking studies revealed the presence of a major receptor-ligand complex of 90 kDa on FHs 738B1 and 160-170 kDa on the other bladder cell lines. All cell lines studied, except RT4, expressed solely FGFR-1. These studies demonstrate that FGF2-saporins have antiproliferative activity on human bladder cancer cell lines. However, the number of high-affinity FGF receptors, and FGF-2 cellular content and secretion are not absolute determinants of cellular sensitivity to FGF2-saporins. PMID- 9344049 TI - Test of two views of impulsivity in hyperactive and conduct-disordered children. AB - The nature of impulsivity in hyperactive and conduct-disordered children was examined in two experiments, one involving a priming task, the other a delayed reaction time task. Four groups of children, aged 7 to 8 years and with IQs in the normal range, were recruited for study: (1) a pure hyperactive group (HA), (2) a hyperactive/conduct-disordered group (HA+CD), (3) a pure conduct-disordered group (CD), and (4) a normal control group (N). When the stimulus configuration and presentation were simple and well organized, none of the three clinical groups displayed any sign of impulsivity at the input/perceptual stage; there was no tendency to rush responding before adequate consideration of the relevant stimuli, i.e. a trading of accuracy for speed. Instead, the HA children were found to be disinhibited at the output/motor stage, i.e. failing to temporarily withhold activated responses. This deficit was found to be specific to the HA children; it was not observed in the CD and HA+CD children. PMID- 9344050 TI - Attention deficits and autistic spectrum problems in children exposed to alcohol during gestation: a follow-up study. AB - Children born to mothers who had abused alcohol throughout pregnancy had severe behavioural and intellectual problems which remained at age 11 to 14 years. Of 24 children examined, 10 had attention deficit hyperactivity disorder (ADHD) with or without developmental coordination disorder, two had Asperger syndrome, and one had an autistic-like condition not meeting the criteria for Asperger syndrome. Six of these 24 attended special schools for the mentally retarded and a further 11 were given special education, leaving only seven attending regular schools without any type of support. The children had difficulties in mathematics, logical conclusions, visual perception, spatial relations, short-term memory, and attention. Sixteen children lived in foster homes. There was a clear correlation between the occurrence and severity of the neuropsychiatric disorder and the degree of alcohol exposure in utero. PMID- 9344051 TI - Adaptive and maladaptive behaviour in children with epileptic encephalopathies: correlation with cerebral glucose metabolism. AB - In the childhood epileptic encephalopathies mental impairment is common and severe. Traditional cognitive assessment is difficult because of the low level of performance, autistic features, and the unpredictable effect of seizures. An alternative is to measure adaptive and maladaptive behaviour using instruments administered to the caregivers. Adults with different types of dementia have characteristic patterns of cortical glucose hypometabolism. Thirty-two children were studied using visual and semiquantitative analysis of 18fluorodeoxyglucose positron emission tomographic (PET) scans. The Vineland Scales and the Conners' Questionnaires were used to assess adaptive and maladaptive behaviour. The mean adaptive behaviour composite score was 37.3+/-15.6; all but one subject had a low adaptive level. A profile of relative strength in socialisation and weakness in daily living skills emerged. Up to two-thirds of children had abnormal behaviour patterns, particularly attention-deficit disorders and hyperactivity. Adaptive and maladaptive behaviour was not related to the presence or absence of focal cortical PET abnormalities. However, adaptive behaviour scores showed an inverse correlation with the degree of metabolic abnormality in the frontal lobes. PMID- 9344052 TI - Effects of intraventricular hemorrhage and hydrocephalus on the long-term neurobehavioral development of preterm very-low-birthweight infants. AB - Measures of intelligence, neuropsychological functions, academic skills, and behavioral adjustment were obtained at school-age from children born preterm with no hydrocephalus (N=29), arrested hydrocephalus (N=19), and shunted hydrocephalus (N=17), and a term comparison group (N=23). Most children also received concurrent neurological examinations and MRI brain scans. Results revealed significantly poorer neurobehavioral development in all four domains in preterm children with shunted hydrocephalus. Despite abnormal MRI findings in virtually all children with arrested hydrocephalus, significant differences between preterm children with arrested hydrocephalus and those with no hydrocephalus were largely in areas involving attentional and academic skills. Preterm children with no hydrocephalus tended to show poorer motor development relative to term children. Neurological abnormalities were restricted to children with spasticity in the arrested (N=2) and shunted (N=10) groups. These results highlight the importance of separating cases according to residual neurological and neuroimaging abnormalities in accounting for variations in the neurobehavioral development of preterm, low-birth-weight infants. PMID- 9344053 TI - Respiration patterns during feeding in Rett syndrome. AB - Feeding problems are common in Rett syndrome in which there are characteristic oropharyngeal abnormalities. This study investigated the ways in which individuals regulated their respiration accordingly, and how this affected their overall feeding ability. Respiration during feeding was studied in 28 individuals, recording nasal airflow, chest and abdominal movements, and swallow sounds. Time to first swallow was defined as that between introduction of liquid/solid on a spoon and the first swallow. Six individuals also had videofluoroscopy with simultaneous respiration monitoring. Results indicated different respiratory patterns according to the time to first swallow and neurological status; the amount of time spent in apnoea was particularly important. Videofluoroscopy showed that apnoeas occurred most often when liquid was delayed in the pharynx, but this could be overcome in subjects with a lower level of disability. The carer's estimate of the time for feeding was significantly related to the time spent in apnoea with liquids. PMID- 9344054 TI - The influence of forearm crutches on pelvic and hip kinematics in children with myelomeningocele: don't throw away the crutches. AB - Gait analysis was performed on 16 children with high-sacral-level myelomeningocele who walked with and without crutches to evaluate the influence of crutches on their unique walking pattern. All of the patients used solid ankle foot orthoses (AFOs). Deviations in coronal and transverse planes improved with assisted walking. The timing of stance phase pelvic depression and the magnitude of stance phase hip abduction improved with crutch walking. Pelvic rotation, which was seven times the normal range of motion during no-crutch walking, decreased to four times normal with crutches. Walking velocity was not significantly different between conditions. The results demonstrated that deviations in pelvic and hip kinematics are related to muscle weakness and improve with crutch use. Crutches enable the patient to transfer some weight bearing to their upper extremities which decreases the demand on weak lower extremity musculature. This allows them to maintain functional ambulation with a closer to normal gait pattern. PMID- 9344055 TI - Relations between family environment and adjustment outcomes in young adults with spina bifida. AB - Thirty-two young adults with spina bifida completed a questionnaire (Family Environment Scale) assessing their perceptions of family social environment while growing up. Additionally, subjects responded to a structured interview addressing their current employment status, residential situation, level of community mobility, and extent of social activity. Multiple logistic regression analyses were used to assess the relation between family environment and adjustment as a young adult. With this limited sample, results indicated that perceived family environment explained variance in employment, community mobility, and social activity as an adult, even beyond that explained by lesion level and intelligence. Regression coefficients showed positive relations between perceived family encouragement of independence and achievement and young adult outcomes. In contrast, perceived moral/religious emphasis of the family and degree of family involvement with intellectual/cultural activities evidenced negative relations with the measures of young adult adjustment. PMID- 9344056 TI - Variable presentation of cerebrovascular disease in monovular twins. AB - We describe twin girls with bilateral cerebrovascular disease. In one child, a diagnosis of moyamoya disease was made after presentation in infancy with an acute hemiparesis; her asymptomatic sibling was found to have significant bilateral cerebrovascular disease after neuropsychological evaluation and assessment with transcranial Doppler ultrasound. Both subjects showed a discrepancy between verbal and performance IQ and deficits on a test of frontal lobe function suggesting that these domains should be targeted in cognitive assessment. Family members of subjects with moyamoya are at risk of cerebrovascular disease. Clinical symptoms do not reliably predict disease and those at risk should be offered screening with non-invasive vascular imaging. PMID- 9344057 TI - A male with fetal valproate syndrome and autism. AB - Fetal valproate syndrome (FVS) is characterized by minor craniofacial anomalies, major organ malformations, and developmental delay. We report on a patient who has a clinical phenotype compatible with both FVS and autism. The presence of an autistic disorder in a previously reported case of FVS and similar findings in our patient suggest that a relation between this known teratogen and autism may exist. PMID- 9344058 TI - Botulinum toxin treatment in cerebral palsy: intervention with poor evaluation? PMID- 9344073 TI - Pudendal nerve somatosensory evoked potentials in paediatrics: maturation aspects. AB - Pudendal nerve somatosensory evoked potentials (PN-SSEPs) were recorded in 21 healthy children (age range: 3.3-13.3 years). The dorsal nerve of the penis/clitoris was stimulated and SSEPs were recorded at spinal L1-D12 and at cortical Cz'-Fz. Morphology, latency and amplitude of the cortical SSEPs were evaluated. A cortical response was obtained in all but two subjects. Cortical SSEPs were broader and less defined in shape in the youngest subjects. There was a progressive shortening of the latency of the P and N components during growth. Spinal responses were obtained only in 6 cases. Nine subjects also underwent tibial nerve stimulation. Pudendal and tibial SSEPs differed in their degree of maturation. PMID- 9344074 TI - A method of uroneurophysiological investigation in children. AB - Characteristics and reproducibility of bulbocavernosus reflex (BCR) and pudendal somatosensory evoked potentials (PSEP) elicited by mechanical stimulation in children were tested. Twenty-five male children aged 5-14 years without uroneurological complaints were enrolled in the study. In addition to electrical stimulation, a specially constructed electromechanical hammer triggered by an oscilloscope was used for mechanical stimulation of distal penis. All responses were detected by surface electrodes. The latencies and amplitudes of averaged as well as latencies of single BCR on single and double electrical stimuli were determined. Mechanical stimulation was described as much less unpleasant than electrical stimulation. Both mechanical/electrical stimulation elicited consistent and reproducible responses in high percentages of children (BCR: average, 80%/71%, single, 94%/100%; PSEP: 96%/96%, respectively). BCR latencies were significantly longer and PSEP amplitudes were significantly higher on mechanical stimulation. The compliance with mechanical was much better than with the electrical stimulation and the former can be recommended for clinical use. The effective mechanical stimulus delivered by a particular mechanical stimulator has a characteristic 'delay' (as to the actual point of triggering the oscilloscope ray) which influences the latency reading of responses; appropriate control data are therefore necessary. PMID- 9344076 TI - Median and tibial nerve somatosensory evoked potentials: middle-latency components from the vicinity of the secondary somatosensory cortex in humans. AB - The topography of the middle-latency N110 after radial nerve stimulation suggested a generator in SII. To support this hypothesis, we have tried to identify a homologous component in the tibial nerve SEP (somatosensory evoked potential). Evoked potentials following tibial nerve stimulation (motor + sensory threshold) were recorded with 29 electrodes (bandpass 0.5-500 Hz, sampling rate 1000 Hz). For comparison, the median nerve was stimulated at the wrist. Components were identified as peaks in the global field power (GFP). Map series were generated around GFP peaks and amplitudes were measured from electrodes near map maxima. With median nerve stimulation, we recorded a negativity with a maximum in temporal electrode positions and 106 +/- 12 ms peak latency (mean +/- SD), comparable to the N110 following radial nerve stimulation. After tibial nerve stimulation the latency of a component with the same topography was 131 +/- 11 ms (N130). Both N110 and N130 were present ipsi- as well as contralaterally. Amplitudes were significantly higher on the contralateral than the ipsilateral scalp for both median (3.1 +/- 2.4 microV vs. 1.7 +/- 1.6 microV) and tibial nerve (1.9 +/- 1.2 microV vs. 0.6 + 1 microV). The topography of the N130 can be explained by a generator in the vicinity of SII. The latency difference between median and tibial nerve stimulation is related to the longer conduction distance (cf. N20 and P40). The smaller ipsilateral N130 is consistent with the bilateral body representation in SII. PMID- 9344075 TI - Somatosensory evoked magnetic fields and potentials following passive toe movement in humans. AB - The somatosensory evoked magnetic fields (SEFs) and evoked potentials (SEPs) following passive toe movement were studied in 10 normal subjects. Five main components were identified in SEFs recorded around the vertex around the foot area of the primary sensory cortex (SI). The first and second components, 1M and 2M, were identified at approximately 35 and 46 ms. Equivalent current dipoles (ECDs) of both 1M and 2M were estimated around SI in the hemisphere contralateral to the movement toe, and were probably generated in area 3a or area 2, which mainly receive inputs ascending through muscle and joint afferents. The large inter-individual difference of 1M and 2M in terms of ECD orientation was probably due to a large anatomical variance of the foot area of SI. The third and fourth components, 3M and 4M, were identified at approximately 62 ms and 87 ms, respectively. They appeared to be a single large long-duration component with two peaks. Since the 3M and 4M components were significantly larger than the 1M and 2M components in amplitude and their ECD location was significantly superior to that of 1M and 2M, we suspected that they were generated in different sites from those of 1M and 2M, probably area 3b or area 4. Four components, 1E, 2E, 3E and 4E, were identified in SEPs, which appeared to correspond to 1M, 2M, 3M and 4M, respectively. The variation observed in the scalp distribution of the primary component, 1E, could be accounted for by the variation of the orientation of ECD of the 1M component. There was a large difference in the waveform of the long latency component (longer than 100 ms) between SEFs and SEPs. The 5E of SEPs was a large amplitude component, but the 5M of SEFs was small or absent. We speculate that this long-latency component was generated by multiple generators. PMID- 9344077 TI - H-reflex changes in the course of amyotrophic lateral sclerosis. AB - In this study the H-reflex and M-wave were evoked in a group of ALS patients, to correlate the findings with the clinical state, and to investigate whether a statistical approach for assessing H-reflex changes in the presence of a constant M-wave could be reproducible and helpful in monitoring the course of amyotrophic lateral sclerosis (ALS). The H-reflex and M-wave from the soleus muscle were evoked at different stimulus strengths in 35 patients with definite ALS during the course of their illness. The mean amplitude of the H-reflexes (H-mean) obtained in different sessions within an established range of mean M-response amplitude (M-mean) was calculated. For each patient, M-mean was made constant across sessions. H-mean showed high reproducibility and two different trends of changes which emerged in a 1 year follow-up within the population: a significant progressive increase and a steady decrease. When grouped on the basis of their H mean trend, the patients did not differ in terms of any clinical variables considered. However, the group with progressive increase of H-mean showed a better prognosis. This study has shown that H-mean is effective in assessing the clinical course of ALS and could be useful in monitoring drug effects during clinical trials. PMID- 9344078 TI - Developmental changes in the event-related EEG theta response and P300. AB - Event-related potentials (ERPs) from 50 children (6-11 years) and 10 adults were elicited by auditory passive, and by rare target and frequent non-target stimuli, and analyzed in the time and frequency domains. The latency of the maximal theta response (or the theta frequency component of the ERP) was evaluated with respect to age and scalp topography effects. The major findings were: (1) The latency of the maximal theta response decreased with increasing age in children, although for each stimulus type and location adults had shorter latencies than the children. (2) The developmental time course of latency reduction depended on the electrode location, with the most prominent reduction occurring at 8 years at Cz, and no differences between children groups obtained for the frontal site. (3) Maximal theta response latency was strongly associated with the latency of the late parietal P400-700 (P3b) component in children. The results suggest that the developmental latency decrease in P300 processes originate from a decrease in the preceding theta-related processes and may reflect a speeding of cognitive stimulus evaluation. PMID- 9344079 TI - Estimate of physiological variability of peak latency in single sweep P300. AB - Among single sweep records of event-related potentials (ERPs), the peak latency of P300, which is one of the most prominent positive peaks in the ERP obtained in the oddball paradigm, may vary depending on the conditions of the subject. In the analysis of characteristics of the variability in the peak latency, it is important to know to what extent the variability of the measured peak latency (measured variability) is actually caused by physiological factors (physiological variability). In our previous study, a method was developed for judging whether the physiological variability really exists or not, and if it does exist, the developed method extracts the physiological variability from the measured variability based on a limited number of single sweep records. In the present study, based on the P300 waveforms which were detected by blinded visual inspection of the ERP data obtained by an auditory oddball paradigm from 12 healthy adults, the physiological variability was shown to exist with a confidence level of 95% for all subjects. Furthermore, its interval estimate was calculated by subtracting noise variability from the measured variability with a confidence level of 80%, and it was found to range from 17 to 57 ms for all subjects. PMID- 9344080 TI - Dynamics of MLAEP changes in midazolam-induced sedation. AB - This study aimed at assessing the effects of midazolam (MDZ) sedation on auditory brainstem (BAEP) and middle latency (MLAEP) evoked potentials in intensive care conditions. Ten ventilated comatose patients were receiving an intravenous MDZ bolus dose (0.2 mg/kg) followed by a 2 h continuous infusion (0.1 mg/kg/h). MLAEPs and BAEPs elicited by clicks (90 dB HL + masking) were simultaneously and continuously monitored during the first 6 h and for 30 min the next morning. We found no effect of MDZ sedation on BAEPs. Only MLAEP components were modified. However, none of the patients presented any total abolition of the MLAEPs. Bolus injection led to very early alteration of cortical responses, beginning after 5 min and lasting almost 1 h (maximum Pa latency increase, 3.1 ms; maximum Pa-Nb amplitude decrease, 46%). During continuous infusion, MLAEPs remained slightly, although significantly, altered (Pa latency, +1.3 ms; Pa-Nb amplitude, 27%). The Nb wave seemed to be modified earlier and to return to normality later than the Pa wave. These findings incite a careful interpretation of MLAEP tracings acquired during the first hour following MDZ bolus injection. If possible, MDZ should be administered as continuous infusion for reliable interpretation of evoked potential changes in intensive care unit, or during surgery. PMID- 9344081 TI - Event-related potentials of the rat during active and passive auditory oddball paradigms. AB - Event-related potentials (ERPs) of the rat were recorded at the frontal, temporal and parietal areas on the skull during active and passive auditory oddball paradigms, and consisted of P1 (12.7-37.7 ms), N1 (40.0-80.6 ms), P2 (91.7-202.7 ms), N2 (183.7-246.7 ms) and P3 (265.7-462.7 ms) components. Topography and relationship to the paradigm and stimulus types were examined, and unique features were found for each component. P1, N1 and N2 were prominent frontally. However, P2 showed maximum amplitude at the parietal area. N2 and P3 were consistently present only for rare stimuli. During the passive paradigm P3 had a tendency to be greater at the parietal area, but during the active paradigm it had a longer latency and a larger amplitude than during the passive paradigm. No significant difference between the recording sites was observed for P3 latency and amplitude during the active paradigm. The relationship to the paradigm and stimulus types indicates that the rat P3 corresponds to that of the human. There are differences, however, in surface distribution of the ERP components between the rat and the human. The topographical characteristics of the rat ERP, which are possibly due to differences in brain architecture and function, should be taken into consideration when the rat is used for ERP research. PMID- 9344082 TI - Magnetic stimulation over the cerebellum in patients with ataxia. AB - We studied 20 patients with ataxia caused by various disorders using magnetic stimulation over the cerebellum. Results were compared with normal values found for 12 normal volunteers. In normal subjects, a magnetic stimulus over the cerebellum reduced the size of responses evoked by magnetic cortical stimulation when it preceded cortical stimulus by 5, 6 and 7 ms. The grand average of the ratios of the areas of conditioned responses at intervals of 5, 6 and 7 ms to those of control responses was designated the average area ratio (5-7 ms). Suppression of motor cortical excitability was reduced or absent in patients with a lesion in the cerebellum or cerebellothalamocortical pathway, but was normal in patients with a lesion in the afferent pathway to the cerebellum. Normal suppression was observed in Fisher's syndrome. The average area ratio (5-7 ms) correlated well with the severity of ataxia in patients with degenerative late onset ataxia. These results are consistent with those for electrical stimulation of the cerebellum reported previously. We conclude that magnetic stimulation over the cerebellum produces the same effect as electrical stimulation even in ataxic patients. This less painful method can be used clinically to clarify the pathomechanisms for ataxia. Two other clinical uses of this technique were that it revealed clinically undetectable cerebellar dysfunction in patients whose extrapyramidal signs masked cerebellar signs, and that the slow progression of ataxia could be followed quantitatively in patients with degenerative late-onset ataxia. PMID- 9344160 TI - Relationship between physical properties and crystal structures of chiral drugs. PMID- 9344161 TI - Specific inhibition of nitric oxide production in macrophages by phosphorothioate antisense oligonucleotides. AB - The effects of antisense oligonucleotides (ODNs) on nitric oxide (NO) production induced by lipopolysaccharide (LPS) were investigated using thioglycollate induced mouse peritoneal macrophages. Antisense phosphorothioate ODNs (S-oligo) corresponding to a sequence in the neighborhood of the AUG initiation codon of a mouse inducible nitric oxide synthase (iNOS) mRNA, which has a G-quartet motif in its antisense sequence, inhibited NO induction in a dose-dependent manner. Antisense phosphodiester ODNs (D-oligo), 5'- and 3'-terminal phosphorothioate modified antisense ODNs and control scramble and missense S-oligos had no such effect. In addition, control nonsense and two mismatched S-oligos, which include G-quartet motif in their sequences, inhibited NO induction to approximately 50% of those in the control. Antisense S-oligo showed the inhibitory effect on NO production by exposure of macrophages to various concentrations of LPS. Western blot analysis using anti-mouse inducible nitric oxide synthase (iNOS) antibody revealed that antisense S-oligo specifically removed an immunoreactive band at 130 kDa. In addition, the results of reverse transcription-polymerase chain reaction (RT-PCR) revealed that the antisense effect originated from a specific reduction of the targeted iNOS mRNA by hybridization with the antisense S-oligo. Furthermore, no ODNs affected beta-actin mRNA and tumor necrosis factor alpha (TNF-alpha) expression in macrophages stimulated by LPS. These findings demonstrated that antisense S-oligo inhibited NO production derived from iNOS expression in macrophages by an antisense mechanism, including the aptameric effect partially mediated by the G-quartet motif. PMID- 9344162 TI - A study of the relationship between the structure and physicochemical parameters of a homologous series of oxprenolol esters at various pH values and temperatures. AB - A number of beta-adrenergic blockers, including timolol and propranolol, are administered in eyedrops for the treatment of glaucoma, but their therapeutic value is limited by a relatively high incidence of cardiovascular and respiratory side effects. Because of poor ocular bioavailability, many ocular drugs are applied in high concentrations, which give rise to both ocular and systemic side effects. Methods to increase ocular bioavailability include (a) the development of drug delivery devices designed to release drugs at controlled rates, (b) the use of various vehicles that retard precorneal drug loss, and (c) the conversion of drugs to biologically reversible derivatives (prodrugs) with increased corneal penetration properties, from which the active drugs are released by enzymatic hydrolysis. A homologous series of aliphatic esters of oxprenolol were synthesized and investigated as potential prodrugs for ocular use. The stability of each O-acyl derivative was investigated in aqueous solutions over the pH range 2.2-9.0 at 37 degrees C. The observed rate constants (k[obs]), shelf-lives (t90), lipophilicities, and Arrhenius parameters were determined for each ester. A study of the relationship between the structure and physicochemical parameters of the homologous series of oxprenolol esters at various pH values and temperatures was made. PMID- 9344163 TI - Mechanistic evaluation of binary effects of magnesium stearate and talc as dissolution retardants at 85% drug loading in an experimental extended-release formulation. AB - The feasibility of producing extended-release matrix tablets with high drug loadings (80-90% w/w) containing a binary combination of magnesium stearate (MS) and talc (T) at different levels as major dissolution retardants was investigated. Matrix tablets were prepared from a granulation containing theophylline, starch, hydroxypropylcellulose, and varying amounts of MS and T. Using a 32 factorial design, the effect of MS and T levels on the physical properties and drug release characteristics of the tablets was evaluated. Response surface analysis showed that the binary combination of MS and T at levels >3% adversely affected both tensile strength and friability. A parabolic relationship was observed for the increase in time required for the release of 50% of the theophylline (t50%) with increased MS levels. Moreover, as the proportion of MS and T was increased, the release profiles became more linear. A combination of 3% MS and T provided both near zero-order release kinetics as well as a coherent matrix structure. Based on model fitting, a release mechanism combining diffusion and matrix erosion/dissolution is proposed. It may be concluded that in the development of controlled-release systems, the binary combination of MS and T at levels exceeding those conventionally used for lubrication can be employed as an inexpensive, low bulk dissolution retardant for formulations with high drug loading. PMID- 9344164 TI - Determination of rate order for degradation of drugs with nonisothermal stability experiment. AB - The inability of ordinary nonisothermal experiments to determine the rate order (n) of drug degradation is discussed on the basis of a theoretical study of simulated nonisothermal data. In ordinary nonisothermal experiments, using either r or sigma(C[experiment] C[compute])2 as the measure of goodness of fit, the rate order cannot be assessed because the same set of c-t data can be well fitted by different combinations of estimates for n and the activation energy (E). Hence, a new nonisothermal heating-and-cooling model is introduced as a revision, in which sigma(C[experiment] C[compute])2 changes significantly with various kinetic models. Therefore, all the kinetic parameters, including the rate order, can be obtained in one nonisothermal stability experiment. Furthermore, the ability to determine rate order is significantly affected by the extent of drug degradation and experimental error, though not by sampling frequency or temperature change. To demonstrate the applicability of the heating-and-cooling model, the stability and rate order of degradation of vitamin C tablets was studied by this method. PMID- 9344166 TI - Pharmacokinetics of ozagrel and its metabolites after intravenous and oral administrations. AB - The pharmacokinetics of ozagrel, a selective thromboxane A2 synthetase inhibitor, and its metabolites (M1 and M2) was investigated in rats. The plasma concentration-time profile of ozagrel was biexponential with a rapid terminal decay (t1/2beta, 0.173 and 0.160 h at doses of 15 and 45 mg/kg, respectively). Metabolites M1 and M2 appeared in plasma immediately after intravenous (iv) dosing of the parent drug. Similar patterns of metabolites were observed in plasma after oral dosing, although concentrations of M2 were higher than those of M1, indicating the metabolic conversion of ozagrel to M2 and M1. However, a saturable first-pass clearance was seen at a high dose (60 mg/kg) of oral ozagrel. When M2 was administered iv, M1 appeared in the circulation system at appreciable levels, providing the first evidence of metabolic conversion of M2 to M1 in the systemic circulation. Ozagrel was partly metabolized to M2 and M1 in rat intestinal mucosa, although the main metabolic site might be in the liver. The results indicate that the metabolic pathway of ozagrel in rats is the conversion of the parent drug to M2 and M1 and the conversion of M2 to M1. PMID- 9344167 TI - Transmucosal, oral controlled-release, and transdermal drug administration in human subjects: a crossover study with melatonin. AB - The effect of oral controlled-release (CR), oral transmucosal (buccal; TMD) and transdermal (TDD) drug delivery systems on plasma concentrations of melatonin (MT) and its principal metabolite in human subjects using a crossover, single dose design was evaluated. Twelve adult male volunteers participated in the study and received all three dosage forms on three separate occasions. All patch dosage forms were removed after 10 h of wear. Plasma concentrations of the parent drug and its metabolite, 6-sulfatoxymelatonin (MT6s) were measured by radioimmunoassay. Between-subject plasma concentrations of MT were very variable following both oral CR and TDD. Use of the oral CR system gave plasma MT profiles in some subjects that were initially similar to physiological levels, but then differed substantially from physiological in the rate of MT offset; in a few subjects, plasma MT levels remained consistently much below normal nocturnal physiological levels. Also, the ratio of metabolite to parent drug by the oral CR route was many times greater than physiological. TDD resulted in a significant delay in systemic drug levels and a gradual decline in drug delivery after patch removal, possibly due to deposition of melatonin in the skin. TDD failed to simulate the physiological plasma profile of MT (rapid achievement of steady state blood levels and rapid decline after removal of the patch; i.e., so-called "square-wave" profile). TMD provided prompt systemic drug levels with less variability than oral CR or TDD delivery. Also, plasma MT levels fell promptly and rapidly after removal of the patch. No indication of mucosal deposition was observed. TMD was able to mimic the physiological plasma profiles of both MT and its principal metabolite. PMID- 9344165 TI - Paracellular diffusion in Caco-2 cell monolayers: effect of perturbation on the transport of hydrophilic compounds that vary in charge and size. AB - We applied the principles of molecular-size-restricted diffusion within a negative electrostatic field of force to follow the changes in the aqueous pore radius of tight junctions (TJs) induced by perturbants and the accompanying influence on the permeation of neutral (urea and mannitol), cationic (methylamine and atenolol), and anionic (formate and lactate) compounds that vary in size. The perturbants included palmitoyl-DL-carnitine (PC), which opens TJs by an unknown Ca++-independent mechanism, and ethyleneglycol-bis-(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), a Ca++ chelator. Mass transfer resistances of the collagen-coated filter support and the aqueous boundary layers were factored out to yield paracellular permeability coefficients (P[P]). As viewed from the P(P) values of urea and mannitol, EGTA exhibited insignificant effects on pore size at low concentrations compared with control, and then caused a dramatic opening of the TJs over a narrow concentration range (1.35-1.4 mM). The P(P) values for urea and mannitol remained constant at >1.4 mM EGTA. However, PC produced dose-dependent responses from O to 0.15 mM that plateaued at >0.15 mM. In general, cations permeated the cellular TJs faster and anions slower than their neutral images. The effects of changes in pore size (4.6 to 14.6 A in effective radius) on the ability of these solutes to permeate the TJs were analyzed by the Renkin molecular sieving function. These studies established an experimental, theoretical, and quantitative template to assess perturbants of the TJ and define the limits, short of detrimental effects, at which the TJs may be sufficiently perturbed for maximal enhancement of permeation of solutes varying in size and charge. PMID- 9344168 TI - Binding, uptake, and transport of hypericin by Caco-2 cell monolayers. AB - The biological evaluation of hypericin in various test models is hampered by its very poor water solubility. In the present study cyclodextrin formulations and liposomal preparations were investigated for improved delivery and solubility of hypericin in aqueous buffer systems. Caco-2 cells, grown to tight monolayers on 96-well tissue culture plates as well as on Transwell polycarbonate filters, were used to study the membrane binding and the epithelial transport of hypericin. Cumulative transport of hypericin, which could not be measured without the use of cyclodextrins, in apical-to-basolateral direction from cyclodextrin-hypericin buffer solutions was 3-5% at 37 degrees C and approximately 0.12% at 4 degrees C after 5 h. After an incubation time of 1 h at 37 and 4 degrees C, 12.7% +/- 2.6% and 6.5% +/- 0.8%, respectively, of hypericin were found to be bound to or taken up by Caco-2 cells. Liposomal formulations markedly increased the solubility of hypericin in Krebs-Ringer buffer, but there was no effect observed on the binding and transport of hypericin delivered by liposomes in the Caco-2 cell model. Due to the fluorescence properties of hypericin, its interaction with the cells could be visualized by confocal laser scanning microscopy. The results indicate that a significant accumulation of the drug in the cell membrane and the cell nucleus membrane takes place. We conclude that hypericin is absorbed through the intestinal epithelium by passive transcellular diffusion and that increasing its solubility by cyclodextrin appears as a promising approach to increase its oral bioavailability for pharmaceutical formulations. PMID- 9344169 TI - Synthesis and in vitro evaluation of potential antichagasic dipeptide prodrugs of primaquine. AB - American trypanosomiasis (Chagas' disease) is an endemic parasitic disease afflicting more than 20 million people in Latin America. Currently, therapy is unsatisfactory and only two drugs are available. Primaquine, an antimalarial drug, has trypanocidal activity. Dipeptide derivatives of primaquine, Phe-Arg-PQ, Lys-Arg-PQ, and Phe-Ala-PQ, were synthesized. The choice of the peptides was based on the primary specificity of cruzipain, the major cysteine proteinase from T cruzi. The prodrugs obtained were tested on the LLC-MK2 cell culture infected with trypomastigotes forms of T. cruzi. Phe-Arg-PQ, Lys-Arg-PQ, and Phe-Ala-PQ were active in all stages. PMID- 9344170 TI - A mechanistic study of griseofulvin dissolution into surfactant solutions under laminar flow conditions. AB - The in vivo dissolution of many poorly soluble drugs is enhanced by the action of surfactants secreted into the upper gastrointestinal (GI) tract. These substances may act by solubilizing individual drug molecules into two separate liquid phases: the free aqueous phase and a micellar phase in which the drug is incorporated into a complex of two or more surfactant molecules. This complex process, micellar solubilization, was the subject of this in vitro study, wherein griseofulvin (gris) dissolution was observed in flowing surfactant solutions. Aqueous solutions of sodium dodecyl sulfate (SDS), an anionic surfactant, were pumped over a gris tablet embedded in a laminar flow device to simulate flow in the human upper GI tract. SDS solutions were well above the critical micellar concentration (cmc approximately 6-7 mM), and flow rates ranged from 4 to 7 mL/min. Gris solubility in premicellar (4 mM), near-micellar (6 mM), and micellar (>6 mM) SDS solutions was also determined. The measured solubility of gris increased linearly with SDS concentrations above the cmc. Drug solubility in SDS concentrations below the cmc was also higher than that in water. Gris diffusion coefficients were measured using pulsed-field gradient NMR spectroscopy. To determine the controlling mechanism for surfactant-enhanced dissolution, a mathematical model was developed. The model solution, an equation for drug dissolution rate, was compared with experimental data to demonstrate that drug transport away from the solid surface is the slow step in the process. Measured gris diffusion coefficients and solubility values were used as constants in the mathematical model solution and were combined to calculate an effective gris diffusion coefficient. Using these experimentally determined properties, model calculated dissolution rates were within 7% of the measured values. As hypothesized, dissolution rates were found to be directly proportional to the transport properties of the system (effective drug diffusion coefficient and fluid flow rate) as well as to the drug solubility. To further verify transport limited dissolution, the measured dissolution rates were found to be proportional to the surrounding medium flow rate to the 1/3 power, as predicted by the model dissolution rate equation. PMID- 9344171 TI - Topical delivery system for tretinoin: research and clinical implications. AB - A novel topical tretinoin gel formulation containing a patented TopiCare Delivery Compound, polyolprepolymer-2 (PP-2), was shown to significantly reduce local irritation relative to a marketed tretinoin gel preparation while maintaining clinical efficacy in the treatment of acne. Several in vitro percutaneous absorption studies were conducted with 0.025% tretinoin as a model compound to determine the possible mechanism of action of PP-2 on drug delivery into and through human cadaver skin. Results of these studies have repeatedly shown that a new topical gel formulation containing PP-2 significantly reduces tretinoin penetration while potentially enhancing epidermal deposition compared with a commercial topical gel preparation at the same tretinoin concentration. These studies further support a mechanism of action whereby PP-2 serves as a retentate for drug delivery by formation of a liquid reservoir of polymer and solubilized drug on the skin surface and in the upper layers of the skin, thereby modifying delivery of tretinoin into and through skin. This reservoir of drug and polymer was established within 15 min after topical application, and tretinoin was shown to be highly associated with PP-2. These in vitro findings provide a model by which a new tretinoin gel formulation containing PP-2 reduces irritation relative to a commercial tretinoin gel while maintaining clinical efficacy in the treatment of acne vulgaris. PMID- 9344172 TI - Improvement of the pulmonary absorption of (Asu1,7)-eel calcitonin by various absorption enhancers and their pulmonary toxicity in rats. AB - The effects of absorption enhancers on the pulmonary absorption of (Asu1,7)-eel calcitonin (ECT) and their pulmonary toxicity were examined by means of in situ pulmonary experiments. The absorption of ECT from the lungs was estimated by its hypocalcemic effect. The pulmonary membrane toxicity of absorption enhancers was evaluated by the leakage of Evans Blue from the plasma into the lungs. In the absence of absorption enhancers, a slight hypocalcemic effect was obtained following intrapulmonary administration of ECT. However, we found significant hypocalcemic effects after the ECT administration with 10 mM n-lauryl beta-D maltopyranoside (LM), 10 mM sodium glycocholate (NaGC), and 10 mM linoleic acid HCO60 (hydrogenated caster oil) mixed micelle (MM). The plasma calcium levels decreased as the amount of LM coadministered with ECT increased. In contrast, 10 mM EDTA did not improve the pulmonary absorption of ECT. Overall, a correlation between the pulmonary absorption of ECT and local toxicity was observed in the presence of these additives. However, 1 mM LM, 10 mM NaGC, and 10 mM MM improve the pulmonary absorption of ECT with low pulmonary toxicity. These findings suggest that the use of these adjuvants would be a useful approach for improving the pulmonary absorption of ECT. PMID- 9344173 TI - A comprehensive model for enrofloxacin to ciprofloxacin transformation and disposition in dog. AB - The pharmacokinetics of enrofloxacin and ciprofloxacin, its major active metabolite, were determined in dog after oral and intravenous administrations of enrofloxacin and intravenous infusion of ciprofloxacin. A comprehensive model of enrofloxacin and ciprofloxacin disposition was constructed to investigate the extent of enrofloxacin to ciprofloxacin transformation and the influence of the hepatic first-pass effect on the parent compound oral bioavailability. Plasma levels were measured using a validated HPLC method. Enrofloxacin and ciprofloxacin plasma concentration data were fitted simultaneously using a set of differential equations describing a six-compartment model (two compartments for each analyte, one for the liver, and one for the intestinal tract); it was assumed that only a fraction of enrofloxacin was metabolized to ciprofloxacin and that this conversion only occurred in the liver. The fitted parameters obtained from the model were used to calculate plasma clearances (0.729 +/- 0.212 L/h/kg for enrofloxacin, 0.468 +/- 0.094 L/h/kg for ciprofloxacin), distribution volumes (2.45 +/- 0.49 L/kg for enrofloxacin, 1.92 +/- 0.33 L/kg for ciprofloxacin), mean residence times (3.47 +/- 0.78 h for enrofloxacin, 4.20 +/- 0.82 h for ciprofloxacin), and the fractions of enrofloxacin metabolized to ciprofloxacin after intravenous and oral administrations of enrofloxacin. It was shown that enrofloxacin was largely metabolized to ciprofloxacin and that the fractions of metabolized enrofloxacin were similar after intravenous and oral administrations of enrofloxacin (40.44 +/- 10.08 and 40.17 +/- 8.33%, respectively), the hepatic first-pass effect being low (7.15 +/- 1.99%). PMID- 9344174 TI - Disposition of the selective alpha1A-adrenoceptor antagonist tamsulosin in humans: comparison with data from interspecies scaling. AB - Tamsulosin-HCl is an alpha1A-adrenoceptor antagonist that is mainly eliminated by metabolism in animals and humans and is highly bound to alpha1-acid glycoprotein in blood plasma. The disposition of the compound (0.4 mg as modified-release granules in a capsule) was determined in male volunteers, using intravenous (iv) infusion of tamsulosin-HCl (0.125 mg over 4 h) as reference treatment for the assessment of absolute oral bioavailability. Disposition parameters of iv tamsulosin in humans was compared with data predicted from animal data by interspecies scaling techniques. Levels after iv dosing in humans showed a biexponential decline, with mean half-lives (+/-SD) of 1.2 +/- 0.6 and 6.8 +/- 3.5 h, respectively. The mean systemic clearance (+/-SD) was low (viz., 48 +/- 24 mL/min). The mean volume of distribution (+/-SD) was rather small (21 +/- 6 L), and was estimated at 16 +/- 4 L in the steady state. The mean absolute oral bioavailability (+/-SD) was approximated at 100 +/- 19%. Systemic clearance in humans was poorly predictable from a logarithmic clearance versus body weight relation of rat, rabbit, and dog data. The prediction improved dramatically (accuracy 213%) when scaling was done with systemic clearance values of unbound drug, and it improved further (accuracy 59%) with the product of unbound clearance and maximum life-span potential. Also, the prediction of volume of distribution improved dramatically (accuracy 81%) after correction for differences in extent of protein binding between species. The terminal disposition half-life of 7.0 h, as predicted after integrating maximum life-span potential and protein binding in scaling of clearance, was very close to the value of 6.8 h established experimentally in humans. The present results with tamsulosin underline the importance of correction for extent of protein binding in allometric scaling of clearance and distribution volume. PMID- 9344175 TI - Evaluation of solute permeation through the stratum corneum: lateral bilayer diffusion as the primary transport mechanism. AB - Solute permeation across human stratum corneum (SC) was examined in terms of the fundamental bilayer transport properties. A mathematical model was developed to describe the macroscopic SC permeation via the interkeratinocyte lipid domain in terms of (i) the structure and dimensions of the SC, and (ii) the microscale lipid bilayer transport properties, which include the bilayer/water partition coefficient, the lateral diffusion coefficient, the interfacial transbilayer mass transfer coefficient, and the intramembrane transbilayer mass transfer coefficient. The relative importance of the diffusive resistances associated with the bilayer transport properties was evaluated with the model and experimental data. Lateral diffusion coefficients in SC lipid bilayers were calculated from 120 human skin permeability measurements, and compared with previously reported measurements made in SC-extracted lipids. Good qualitative and quantitative agreement was observed, indicating that, in the context of the model, the diffusive resistance associated with lateral diffusion is sufficient to explain the overall resistance of solute permeation through the SC. A similar analysis shows that the diffusive resistance associated with interfacial transbilayer transport is not capable of explaining the experimental permeation values, thus supporting this finding. The lateral diffusion analysis also revealed a bifunctional size dependence of transport within the SC, with a strong size dependence for small solutes (<300 Da) and a weak size dependence for larger solutes. PMID- 9344176 TI - Prediction of distribution coefficient from structure. 2. Validation of Prolog D, an expert system. AB - Prolog D is a program that formalizes, in a controllable and reproducible manner, an algorithm developed to predict distribution coefficients of ionizable compounds at a given pH and varying counterion concentrations. Its predictive power has been evaluated with experimental log D values measured under standard conditions of buffers and ionic strength. Calculations were performed with the three different options for the estimation of partition coefficients (log P) implemented in the program. Considering the diversity of test compounds as well as the present state of the art in log P and pKa predictions, Prolog D proved to be very efficient and can be used as a tool to provide lipophilicity data. Prediction patterns and correlations with the observed data are of almost equal quality for all options, permitting acceptable results for 80% of the data. PMID- 9344177 TI - Ocular penetration and bioconversion of prostaglandin F2alpha prodrugs in rabbit cornea and conjunctiva. AB - The objective of this study was to identify prostaglandin F2alpha (PGF2alpha) prodrugs that have an optimal ocular absorption profile and therefore could be potentially useful for the treatment of glaucoma. Rabbit cornea, conjunctiva, and iris/ciliary body were mounted in a flow-through chamber to evaluate the permeability and bioconversion of PGF2alpha and its prodrugs. The prodrugs tested were PGF2alpha 1-isopropyl, 1,11-lactone, 15-acetyl, 15-pivaloyl, 15-valeryl, and 11,15-dipivaloyl esters. After 4 h in the donor or acceptor compartments, the products and formation of PGF2alpha were analyzed by HPLC. Effects on intraocular pressure and ocular surface hyperemia were also determined. All prodrugs penetrated the rabbit cornea faster than PGF2alpha by 4- to 83-fold. All prodrugs penetrated conjunctiva faster than PGF2alpha, except the 15-acetyl ester prodrug, which was equally permeable. No direct correlation between drug lipophilicity and permeability across the cornea or conjunctiva was apparent. The most metabolically stable prodrug was the 1,11-lactone, followed by the 11,15 dipivaloyl, 15-pivaloyl, 15-acetyl, 1-isopropyl, and the 15-valeryl esters, the latter of which was extensively converted to PGF2alpha. A separation index for various prodrugs was calculated from the ratio of the bioavailable PGF2alpha for ocular hypotension to the bioavailable PGF2alpha for hyperemia. The highest separation index was observed for the 1,11-lactone prodrug (2.33), followed by the 11,15-dipivaloyl ester prodrug (1.80). Thus the 1,11-lactone and 11,15 dipivaloyl ester prodrugs appeared to be superior to the others in providing bioavailable PGF2alpha for ocular hypotension, while minimizing hyperemia. The favorable separation index for these compounds appeared to be due to their metabolic stability at the corneal surface and conjunctiva combined with sufficient bioavailability for ocular hypotension. PMID- 9344178 TI - Correlation between log P and ClogP for some steroids. PMID- 9344179 TI - An explanation for the variation of the sonophoretic transdermal transport enhancement from drug to drug. AB - Over the last few decades, application of therapeutic ultrasound (frequency between 1 and 3 MHz and intensity between 1 and 2 W/cm2) has been attempted to enhance transdermal transport of several drugs, a method referred to as sonophoresis. The sonophoretic enhancement of transdermal drug transport was found to vary significantly from drug to drug. In certain cases, ultrasound did not induce any enhancement of transdermal drug transport. This variation in the efficacy of sonophoresis has raised a controversy regarding its applicability as a transdermal delivery enhancer. The objective of this paper is to provide a summary of the literature data on sonophoresis and an explanation for the observed variation of the sonophoretic enhancement from drug to drug. This paper also presents an equation to qualitatively predict whether therapeutic ultrasound may enhance transdermal transport of a given drug based on knowledge of the drug passive skin permeability and octanol-water partition coefficient. PMID- 9344180 TI - Ultrastructural plasticity of excitatory synapses. PMID- 9344183 TI - Useless or helpful? The "limbic system" concept. AB - The "limbic system" is a widespread and frequently used concept in the neurosciences although it is characterized by many different and often vague or even contradictory definitions. This concept has therefore received an increasing amount of critique during the past years. An appraisal of various opinions - ranging from rejection to limited use and support - led us to suggestions concerning its appropriate use. Most importantly, the diversity of definitions and the weak empirical foundation require careful consideration as to what extent certain aspects of the "limbic system" concept can be useful heuristic tools in scientific reasoning. PMID- 9344185 TI - Management of psoriasis with calcipotriol used as monotherapy. AB - BACKGROUND: The vitamin D analog calcipotriene/calcipotriol (Dovonex/Daivonex) offers advantages over other forms of topical therapy in some patients with psoriasis. OBJECTIVE: We review the studies of the use of calcipotriol alone in the management of psoriasis. METHODS: The literature concerning topical calcipotriol therapy was reviewed. RESULTS: Calcipotriol compares well with other standard forms of topical therapy for psoriasis. Irritation of the skin may occur but is generally mild. Treatment can often be restarted after the irritation has cleared. CONCLUSION: Treatment with calcipotriol ointment, cream, or solution is effective and safe in many patients with psoriasis. PMID- 9344182 TI - A critical review of 5-HT brain microdialysis and behavior. AB - Serotonin (5-HT) has been implicated in many central nervous system-mediated functions including sleep, arousal, feeding, motor activity and the stress response. In order to help establish the precise role of 5-HT in physiology and behavior, in vivo microdialysis studies have sought to identify the conditions under which the release of 5-HT is altered. Extracellular 5-HT levels have been monitored in more than fifteen regions of the brain during a variety of spontaneous behaviors, and in response to several physiological, environmental, and behavioral manipulations. The vast majority of these studies found increases (30-100%) in 5-HT release in almost all brain regions studied. Since electrophysiological studies have shown that behavioral arousal is the primary determinant of brain serotonergic neuronal activity, we suggest that the increase in 5-HT release seen during a wide variety of experimental conditions is largely due to one factor, namely an increase in behavioral arousal/motor activity associated with the manipulation. PMID- 9344181 TI - Short-term plasticity in thalamocortical pathways: cellular mechanisms and functional roles. AB - Information reaches the neocortex through different types of thalamocortical pathways. These differ in many morphological and physiological properties. One interesting aspect in which thalamocortical pathways differ is in their temporal dynamics, such as their short-term plasticity. Primary pathways display frequency dependent depression, while secondary pathways display frequency-dependent enhancement. The cellular mechanisms underlying these dynamic responses involve pre- and post-synaptic and circuit properties. They may serve to synchronize, amplify and/or filter neural activity in neocortex depending on behavioral demands, and thus to adapt each pathway to its specific function. PMID- 9344184 TI - Vitamin D: a calciotropic hormone regulating calcium-induced keratinocyte differentiation. AB - BACKGROUND: Most biologically active forms of vitamin D3 (1,25(OH)2D3) and its analogs have functions and therapeutic potential that extend beyond those of regulating bone mineralization and intestinal calcium transport. OBJECTIVE: We attempt to provide the rationale for the effectiveness of 1,25(OH)2D3 and its analogs in dermatology. METHODS: The recent literature on the mechanisms of action of 1,25(OH)2D3 were reviewed. RESULTS: 1,25(OH)2D3 affects keratinocyte differentiation partly through its regulation of epidermal responsiveness to calcium. Calcium and 1,25(OH)2D3 interact in transcription of the genes required for differentiation and in the stability of the mRNAs produced from such genes. CONCLUSION: An understanding of these mechanisms provides a rationale for treatment of various skin diseases with 1,25(OH)2D3 and its analogs and directs development of more effective therapy. PMID- 9344186 TI - Topical application of calcipotriene and corticosteroids: combination regimens. AB - BACKGROUND: Side effects of topical corticosteroids limit their long-term use. Calcipotriene/calcipotriol (Dovonex/Daivonex) ointment is not associated with any of the side effects of corticosteroids and has been shown to thicken the skin in contrast to the cutaneous atrophy caused by topical steroids. OBJECTIVE: We attempted to determine whether the addition of calcipotriene to a regimen of topical steroids results in an improved benefit/risk ratio. METHODS: Published and unpublished data on combination regimens were reviewed. RESULTS: In long-term regimens for psoriasis, substituting calcipotriene for topical corticosteroids may result in a steroid-sparing effect. Conversely, topical corticosteroids may suppress the development of local cutaneous irritation that occurs in patients treated with calcipotriene ointment. CONCLUSION: Psoriasis regimens combining calcipotriene ointment with superpotent steroids such as halobetasol ointment can result in greater improvement and fewer side effects. PMID- 9344188 TI - Combined topical calcipotriene ointment 0.005% and various systemic therapies in the treatment of plaque-type psoriasis vulgaris: review of the literature and results of a survey sent to 100 dermatologists. AB - BACKGROUND: Plaque-type psoriasis may at times require systemic therapy. There are limited data as to whether topical calcipotriene ointment 0.005% can be used to increase the efficacy and improve the risk/benefit ratio of concurrent systemic antipsoriatic therapy. OBJECTIVE: We attempt to answer this question by means of a literature review and results of a written survey that was sent to 100 international psoriasis treatment experts. METHODS: The survey was sent to academic and psoriasis treatment center-based dermatologists who treat approximately 3000 to 4000 patients with psoriasis per month. The survey requested that dermatologists relate their experience regarding the safety and efficacy of topical, systemic, and combined topical/systemic agents in psoriasis after 8 weeks of therapy. RESULTS: The results of the survey support the experience in the literature regarding the favorable use of calcipotriene ointment combined with systemic therapy for the treatment of psoriasis. CONCLUSION: Combination therapy with calcipotriene ointment and acitretin/etretinate, cyclosporine, methotrexate, or phototherapy usually enhances efficacy while improving the risk/benefit ratio by decreasing exposure to the potentially hazardous systemic agent. PMID- 9344187 TI - Calcipotriol/calcipotriene (Dovonex/Daivonex) in combination with phototherapy: a review. AB - BACKGROUND: Calcipotriol/calcipotriene (Dovonex/Daivonex) ointment plus phototherapy has been used in the treatment of psoriasis. OBJECTIVE: We will attempt to clarify two issues: First, is there any benefit to combining the above treatment modalities? Second, is there any increased risk associated with the use of the combination? METHODS: A complete review of the literature revealed four studies dealing with the combination of calcipotriol with UVB phototherapy and three studies dealing with the combination of calcipotriol with PUVA phototherapy. RESULTS: These studies showed an advantage to the use of the combination compared with the use of either treatment alone. CONCLUSION: Topical calcipotriol enhances the effect of UVB and PUVA phototherapy. PMID- 9344190 TI - The future of vitamin D in dermatology. AB - BACKGROUND: Vitamin D3 analogs have been found to be effective in treating psoriasis. OBJECTIVE: We attempted to identify the targets and actions of vitamin D3 in the skin and to explore the availability of synthetic vitamin D3 analogs with selective actions on particular cell types or cell functions. METHODS: A review of the literature focused on the cellular targets of vitamin D3 in the skin and within the immune system. Furthermore, the use of novel vitamin D3 analogs in skin diseases other than psoriasis was reviewed. RESULTS: The vitamin D receptor has been detected in most skin cells, which means that keratinization, hair growth, melanogenesis, fibrogenesis, angiogenesis, and immune-mediated processes are potential targets for vitamin D3. Vitamin D3 analogs have been synthesized with a higher therapeutic index or a higher degree of selectivity than the natural form of vitamin D3. CONCLUSION: Vitamin D3 analogs with wide ranging clinical applications may become available for dermatology. PMID- 9344189 TI - The use of topical calcipotriene/calcipotriol in conditions other than plaque type psoriasis. AB - BACKGROUND: Topical calcipotriene ointment has been approved for the treatment of plaque-type psoriasis. OBJECTIVE: This article explores the possible use of topical calcipotriene ointment in the treatment of nail and intertriginous psoriasis, palmoplantar and pustular psoriasis, Reiter's syndrome, pityriasis rubra pilaris, and disorders of keratinization. METHODS: The recent literature is reviewed. RESULTS: Recent reports suggest that certain ichthyoses (particularly the hyperproliferative variants) and keratodermas may respond to topical calcipotriene ointment. The activity of calcipotriene relates to a dose-dependent decrease in proliferation and an increase in terminal differentiation of keratinocytes. CONCLUSION: Patients with other disorders characterized by epidermal hyperproliferation may also be candidates for treatment. The use of calcipotriene in treating congenital hyperproliferative disorders is limited by the theoretical risk of hypercalcemia from absorption of the drug after application to extensive areas of skin. PMID- 9344191 TI - Cutaneous vascular anomalies. Part I. Hamartomas, malformations, and dilation of preexisting vessels. AB - Classification of cutaneous vascular anomalies is difficult because conceptual confusion persists between vascular neoplasms and malformations. However, hemangiomas of the infancy fulfill criteria both for hyperplasia and neoplasm because they result from proliferation of endothelial cells, but often undergo complete regression. Despite these pitfalls we have classified cutaneous vascular anomalies into the following categories: hamartomas, malformations, dilatations of preexisting vessels, hyperplasias, benign neoplasms, and malignant neoplasms. In this first part of our clinicopathologic review of vascular anomalies, hamartomas, malformations, and dilatation of preexisting vessels are covered. Hamartomas include several combined vascular and melanocytic proliferations grouped as phakomatosis pigmentovascularis and the so-called eccrine angiomatous hamartoma that consists of proliferations of both eccrine glands and blood vessels. Vascular malformations result from anomalies of embryologic development, and in some of them the abnormalities of the involved vessels are more functional than anatomic, as is the case of nevus anemicus. In contrast, other cutaneous vascular malformations show striking morphologic abnormalities of the vascular structures. These anatomic vascular malformations are subdivided into the following groups: capillary, venous, arterial, lymphatic, and combined anomalies. Spider angioma, capillary aneurysm-venous lake, and telangiectases are not vascular proliferations at all, but dilations of preexisting vessels. In our opinion, most of the lesions described with the generic term of "angiokeratoma" are not authentic vascular neoplasms, but hyperkeratotic malformations of capillaries and venules or acquired telangiectases of preexisting blood vessels of the papillary dermis. Therefore the first group of these "angiokeratomas" are included in the vascular malformations section, and the second group are covered in the section of dilation of preexisting vessels. Lymphangiectases are considered the lymphatic counterpart of angiokeratomas because they result from ectasia of preexisting lymphatic vessels of the papillary dermis. PMID- 9344192 TI - Cutaneous reactions to recombinant human interferon beta-1b: the clinical and histologic spectrum. AB - BACKGROUND: Recombinant human interferon beta-1b has been recently approved for the treatment of multiple sclerosis. A significant proportion of patients treated with this medication experienced cutaneous reactions. OBJECTIVE: We describe the clinical and histologic features of cutaneous reactions to recombinant human interferon beta-1b. METHODS: Consecutive patients with cutaneous reactions to recombinant interferon beta-1b were evaluated clinically and by biopsy. RESULTS: Clinical lesions varied from subtle uninflamed sclerotic dermal plaques to erythematous plaques to cutaneous ulcers at injection sites. The nonsclerotic lesions were frequently painful. The firm plaques showed fibrosis histologically, whereas nonsclerotic inflammatory lesions demonstrated a consistent pattern of vascular thrombosis. Hematologic evaluation demonstrated platelet activation in most patients with inflammatory lesions, a feature also noted before interferon treatment in some patients. CONCLUSION: Therapy with recombinant interferon beta 1b is associated with a spectrum of cutaneous reactions and vascular thrombosis. PMID- 9344193 TI - Influence of UVB therapy on dermoscopic features of acquired melanocytic nevi. AB - BACKGROUND: Exposure to UV radiation can lead to clinical, histologic, and ultrastructural changes in acquired melanocytic nevi. OBJECTIVE: We investigated whether UVB therapy can induce changes in melanocytic nevi detectable by dermoscopy. METHODS: Eighty acquired melanocytic nevi of 13 patients (10 females, 3 males; mean age, 28 years; range, 13 to 62 years) undergoing UVB therapy were documented under standardized conditions by means of a Dermaphot apparatus before and at the end of suberythemal UVB therapy. The mean duration of therapy was 8 weeks (range, 2 to 17 weeks) and the mean total UVB dose was 1120 mJ/cm2 (range, 247 to 2771 mJ/cm2). During UV irradiation, 40 nevi were left unprotected and 40 nevi were protected from UV exposure in a randomized manner. Color dermoscopic images of nevi before and after UVB therapy were projected side by side and examined blindly by five investigators. Fifteen different features were evaluated in the nevi. RESULTS: Unprotected nevi became more irregular (p < or = 0.01) and darker brown (p < or = 0.03) by the end of the therapy, whereas the protected nevi showed no significant changes. CONCLUSION: Suberythemal UVB therapy can lead to changes in the dermoscopic image of acquired melanocytic nevi, presumably by activating melanocytes. PMID- 9344194 TI - The economic impact of psoriasis increases with psoriasis severity. AB - BACKGROUND: Psoriasis treatments are known to be costly, but little is known about the financial impact of psoriasis and the way in which it relates to the severity of the disease. OBJECTIVE: This study was performed to obtain an estimate of the treatment costs faced by patients with psoriasis. METHODS: A total of 578 anonymous mail surveys were distributed to patients with psoriasis; 318 surveys were returned (55%). Psoriasis severity was assessed with the previously validated Self-Administered Psoriasis Area Severity Index (SAPASI). RESULTS: The total and out-of-pocket expenses to care for psoriasis were correlated with psoriasis severity (r = 0.26, p = 0.0001). There were no sex (p = 0.9) or racial (p = 0.4) differences in total expenditures. Severity was correlated with how bothersome to the patient was the cost of treatment (r = 0.30, p = 0.0001), the time required for treatment (r = 0.38, p = 0.0001), and the time lost from work (r = 0.23, p = 0.0001). Lower quality of life at work and in money matters also correlated with severity of psoriasis. Higher family income was associated with less time spent caring for psoriasis and less interference with work around the home. CONCLUSION: As expected, the expenses caring for psoriasis are greater for patients with more severe disease. These costs and other financial implications are associated with lower quality of life for patients with more severe psoriasis. PMID- 9344195 TI - Prevalence of acne keloidalis nuchae in football players. AB - BACKGROUND: Acne mechanica (AM) is common in football players. Severe cases of nuchal AM may precede acne keloidalis nuchae (AKN). Factors that may be associated with the progression of nuchal AM to AKN are unknown. The prevalence of AKN in football players has not been reported. OBJECTIVE: We investigated the frequency of nuchal AM and AKN within a susceptible population and attempted to identify factors that may be associated with AKN. METHODS: Four hundred fifty three high school, collegiate, and professional football players were examined for the presence of nuchal AM or AKN. Those with positive findings completed a questionnaire regarding their disease. RESULTS: Nuchal AM was more prevalent in high school players (15.5%) than older players (1.2%). AKN was more frequent in players beyond the high school level and was found exclusively in blacks. AKN was not associated with a positive family history of AKN nor a positive personal or family history of keloid formation. CONCLUSION: AKN occurs almost exclusively in blacks. The level of football play may be associated with the development of AKN. Positive family history of AKN and positive family or personal history of keloids is not associated with AKN development. PMID- 9344197 TI - The independent pathology laboratory as a reporting source for cutaneous melanoma incidence in Iowa, 1977-1994. AB - BACKGROUND: Health care changes during the past decade have resulted in a greater proportion of cutaneous melanoma (CM) cases diagnosed in nonhospital settings, increasing the potential for cases to be missed by population-based cancer registries. OBJECTIVE: Our purpose was to assess changes in case-finding sources in Iowa from 1977 to 1994 and to determine the extent of underreporting for the State Health Registry of Iowa, a population-based cancer registry. METHODS: This study examines changing trends in the incidence of CM and compares case-finding sources (hospitals/clinics, hospital pathology laboratories, and independent pathology laboratories). A survey of dermatologists serving Iowans provides estimates of underreporting. RESULTS: During the period 1977 to 1994, invasive CM increased 82%, whereas in situ CM increased 900%. The proportion of CM cases diagnosed in independent pathology laboratories increased to 25% of all cases. A range of 10.4% to 17.1% underreporting was estimated based on the survey of dermatologists. CONCLUSION: To improve the accuracy of surveillance, population based cancer registries need to make a greater effort accessing pathology reports from nonhospital settings. PMID- 9344196 TI - Expression of the human erythrocyte glucose transporter Glut1 in cutaneous neoplasia. AB - BACKGROUND: The increased glucose uptake seen in cancer cells correlates with the expression of human erythrocyte glucose transporter (Glut1) protein in certain human malignancies. OBJECTIVE: Our purpose was to determine Glut1 expression in cutaneous neoplasms. METHODS: A polyclonal anti-Glut1 antibody (MYM) and a standard ABC immunoperoxidase technique were used to determine Glut1 expression in invasive squamous cell carcinomas (SCCs), SCC in situ, basal cell carcinomas (BCCs), melanomas, actinic keratoses (AKs), seborrheic keratoses, common acquired nevi, and scars with regenerative epidermal hyperplasia. RESULTS: All of the cases of SCC in situ, 14 of 15 (93%) of the SCC, and 13 of 15 AKs (87%) showed intense membranous staining for Glut1. Glut1 staining was present in the epidermis of 8 of 15 scars (53%) but was not detected in any BCC, even in areas of focal keratinization and squamous metaplasia. Glut1 reactivity was absent in the melanomas and seborrheic keratoses. CONCLUSION: Glut1 expression in a cutaneous lesion strongly suggests a proliferative lesion of the squamous cell type. PMID- 9344198 TI - Major suppression of pro-alpha1(I) type I collagen gene expression in the dermis after keloid excision and immediate intrawound injection of triamcinolone acetonide. AB - BACKGROUND: A keloid is a benign tumor that contains excess collagen, primarily type I collagen. A common therapy is intralesional injection of a glucocorticosteroid, such as triamcinolone acetonide (TA). Surgical excision is also common; often a glucocorticosteroid is injected weeks after excision when wound repair has already begun. OBJECTIVE: Our purpose was to determine the efficacy of TA in reducing the pro-alpha1(I) type I collagen mRNA in the dermis, when TA is injected into the wound bed immediately after surgical excision of the keloid. METHODS: Six patients with previously untreated keloids were studied. Three were treated with 10 mg/ml of TA immediately after excision of the keloid (experimental group); the other three patients were not treated with TA until 2 weeks after excision (control). Punch biopsy specimens were obtained from the TA treated sites 2 weeks after removal of the keloid and from the wounds of the control group of patients before they were treated with TA. Sections were prepared for in situ hybridization analysis of the pro-alpha1(I) collagen mRNA, as well as for histochemical analysis of collagen fibers. RESULTS: All keloids showed greatly elevated levels of pro-alpha1(I) type I collagen mRNA in the dermis. Postsurgical wounds injected with TA after removal of the keloid expressed decreased pro-alpha1(I) collagen transcripts, compared with skin not treated with TA. The collagen bundles were also thinner and less dense in the TA treated skin. CONCLUSION: Downregulation of the type I collagen gene expression is elicited by immediate TA injection after keloid excision. This suggests that prevention of recurrent keloid growth is possible if surgical excision is accompanied by immediate TA injection into the wound bed and that healing of the wound is not apparently compromised by inhibition of type I collagen gene expression. PMID- 9344200 TI - Intralesional bleomycin-mediated electrochemotherapy in 20 patients with basal cell carcinoma. AB - BACKGROUND: A new anticancer therapy, electrochemotherapy (ECT), has been introduced that entails exposing cancerous tissues to short pulses of electricity during chemotherapy. This enhances cell membrane permeability and has been shown to have potent antitumor effects in vitro in animal models and in several clinical trials, including nevoid basal cell carcinoma (BCC). OBJECTIVE: We report the effects of ECT on 20 patients with primary BCC. METHODS: Electrical pulses were delivered to 54 tumors after administration of intralesional bleomycin sulfate. RESULTS: Complete responses were observed in 53 (98%), and in the majority of these (94%) after a single treatment. No recurrences have been recorded with a mean of 18 months of observation. CONCLUSION: Although these are preliminary results, ECT appears to be an effective alternative to surgical excision for the treatment of primary BCC. PMID- 9344199 TI - Treatment of acne with a combination clindamycin/benzoyl peroxide gel compared with clindamycin gel, benzoyl peroxide gel and vehicle gel: combined results of two double-blind investigations. AB - BACKGROUND: It has previously been shown that a combination of erythromycin and benzoyl peroxide is superior to either ingredient when used alone in the treatment of acne. A clindamycin/benzoyl peroxide combination gel might have an advantage over erythromycin/benzoyl peroxide gel because the former does not require refrigeration after it is dispensed. OBJECTIVE: Our purpose was to determine the efficacy and safety of a combination clindamycin/benzoyl peroxide gel when compared with benzoyl peroxide, clindamycin, or vehicle gels. METHODS: In two double-blind, randomized, parallel, vehicle-controlled trials, patients were treated for 11 weeks with once-nightly application of one of the above preparations. Evaluations were performed at 2, 5, 8, and 11 weeks and included lesion counts and assessment of global responses and irritant effects. RESULTS: A total of 334 patients completed the study. All three active preparations were significantly superior to the vehicle in global improvement and in reducing inflammatory lesions and noninflammatory lesions. The combination gel was significantly superior to the two individual agents in global improvement and reduction of inflammatory lesions and also to the clindamycin gel in reducing noninflammatory lesions. There was no significant difference in tolerance to the active gels versus the vehicle gel. CONCLUSION: In the treatment of acne, topical clindamycin/benzoyl peroxide combination gel is well tolerated and superior to either individual ingredient. PMID- 9344201 TI - Mohs micrographic surgery for the treatment of dermatofibrosarcoma protuberans. Results of a multiinstitutional series with an analysis of the extent of microscopic spread. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is an uncommon soft-tissue tumor of the skin; its microscopic extent of invasion beyond the grossly visible tumor is frequently difficult to appreciate. Although wide local excision has been the standard treatment of DFSP, recurrence rates range from 11% to 53%. Because Mohs micrographic surgery allows the extent of excision to be tailored to the microscopic extent of tumor, we evaluated this technique for the treatment of primary and recurrent DFSP. OBJECTIVE: Our purpose was to determine the local recurrence rate and microscopic extent of spread of primary and recurrent DFSP after treatment with Mohs micrographic surgery. METHODS: The records of 58 patients with primary and recurrent DFSP treated with Mohs micrographic surgery at three institutions were reviewed and the macroscopic and microscopic extents of tumor were recorded. RESULTS: One patient with a twice-recurrent DFSP had another recurrence after Mohs micrographic surgery, for an overall local recurrence rate of 2% (zero for primary tumors and 4.8% for recurrent tumors). There were no cases of regional or distant metastases. Macroscopic tumor size ranged from 0.3 x 0.6 cm to 30 x 20 cm, whereas microscopic (postoperative) size ranged from 1.8 x 1.0 cm to 35 x 40 cm. We calculated the likelihood that a given width of excision around the macroscopic tumor would clear the entire microscopic extent of tumor. Standard wide excision with a width of 1 cm around the primary tumor would have left microscopic residual tumor in 70.7%; a width of 2 cm, 39.7%; 3 cm, 15.5%; and 5 cm, 5.2%. Even an excision width of 10 cm would not have cleared the microscopic extent of some tumors, despite taking a huge excess of normal tissue. CONCLUSION: Treatment of primary and recurrent DFSP by Mohs micrographic surgery results in a low recurrence rate because of the ability of the technique to permit the detection and excision of microscopic tumor elements in even the most asymmetric tumors. Whatever type of surgery is chosen to treat DFSP, it is necessary to assess the entire perimeter of the tumor for microscopic extension and to achieve tumor-free margins in all directions. PMID- 9344202 TI - The staged cheek-to-nose interpolation flap for reconstruction of the nasal alar rim/lobule. AB - BACKGROUND: A deep defect of the nasal alar rim or lobule may represent a unique and difficult challenge because of the lax free margin and structural support supplied by the alar rim and lobule. Traditional closure strategies, including granulation, full thickness skin grafting, or nasolabial transposition flaps may result in unsatisfactory cosmetic and functional outcomes. OBJECTIVE: This article describes our experience with the staged cheek-to-nose interpolation flap for repairing deep skin cancer excision defects of the nasal alar rim and lobule. METHODS: The staged cheek-to-nose interpolation flap was used immediately after Mohs micrographic surgery to repair 18 deep nasal alar rim/lobule defects. In 13 patients, a free cartilage graft was used to restore structural support. RESULTS: The cosmetic and functional outcomes of each repair were judged from good to excellent by patient and surgeon. No cases of infection or flap necrosis occurred. To enhance the cosmetic outcome, three patients underwent spot dermabrasion within 2 months after flap detachment. CONCLUSION: The staged cheek to-nose interpolation flap, with or without free cartilage grafts, consistently provides good to excellent cosmetic and functional outcomes when performed on properly selected deep nasal alar rim/lobule defects. PMID- 9344203 TI - The immune response in halo nevi. AB - The mechanism(s) responsible for halo nevus presents a provocative link with the immune response to melanoma. Although no direct demonstration of melanocyte killing has been observed by the immune effector cells found within the halo, the abundance of antigen-presenting cells in the regressing nevus and the presence of T lymphocytes at the site of depigmentation suggest that these cells participate in the halo phenomenon. Within the latter population of cells, evidence points to the involvement of CD8+ T cells as potential effectors in the destruction of nevomelanocytes. The break in tolerance that triggers migration and the presumed activation of these and other lymphocytes in the nevus in the apparent absence of disease remains unexplained. This brief overview reviews the evidence for the participation of the immune response in the genesis of the halo nevus. PMID- 9344204 TI - Guidelines of care for neurofibromatosis type 1. American Academy of Dermatology Guidelines/Outcomes Committee. PMID- 9344205 TI - Guidelines of care for hemangiomas of infancy. American Academy of Dermatology Guidelines/Outcomes Committee. PMID- 9344206 TI - Surgical pearl: suction syringe for epidermal grafting. PMID- 9344208 TI - Successful treatment of a persistent radiation ulcer by low power laser therapy. PMID- 9344207 TI - Sinus histiocytosis with massive lymphadenopathy: presentation as giant granuloma annulare and detection of human herpesvirus 6. PMID- 9344209 TI - Resolution of Kaposi's sarcoma associated with undetectable level of human herpesvirus 8 DNA in a patient with AIDS after protease inhibitor therapy. PMID- 9344210 TI - Lymphatic mapping for Merkel cell carcinoma. PMID- 9344212 TI - Terbinafine-induced acute generalized exanthematous pustulosis confirmed by a positive patch-test result. PMID- 9344211 TI - An unusual cause of pain after nevus excision: complex regional pain syndrome. PMID- 9344213 TI - Acquired progressive lymphangioma. PMID- 9344214 TI - Psoralen UVA therapy for linear and generalized morphea. PMID- 9344215 TI - Symmetric polyarthritis with livedo reticularis: a newly recognized manifestation of hepatitis C virus infection. PMID- 9344217 TI - Amelanotic melanoma in a black man. PMID- 9344216 TI - Antimicrobial effects of lidocaine, bicarbonate, and epinephrine. PMID- 9344218 TI - Medical student publications: a faculty mentor's perspective. PMID- 9344219 TI - Primary care in dermatology. PMID- 9344220 TI - Dermatologists as primary care givers. PMID- 9344221 TI - HIV-associated eosinophilic pustular folliculitis. PMID- 9344222 TI - Long-term low-dose cyclosporine therapy for psoriasis. PMID- 9344223 TI - Reliability of acne lesion counting. PMID- 9344224 TI - Developmental changes in pharmacokinetics of recombinant human insulin-like growth factor-I in rats. AB - Developmental changes in pharmacokinetics of recombinant human insulin-like growth factor-I (rhIGF-I) were investigated after i.v. administration to rats aged 4 and 7 weeks, as young growing rats and adult rats, respectively. rhIGF-I in the plasma declined multi-exponentially in both groups of rats. Plasma concentrations of rhIGF-I were lower at almost all the time points examined in 4 weeks old rats than 7 weeks old rats. The values of total body clearance (CL[total]) and mean residence time (MRT) indicated that rhIGF-I disappeared more rapidly in 4 weeks old rats than 7 weeks old rats at any dosage. Dose-dependent pharmacokinetics was observed in 7 weeks old rats: the higher the dosage was, the larger the value of CL(total) came to be, but not in 4 weeks old rats. The amounts of IGF-binding proteins (IGFBPs) in the plasma were assessed by determining the endogenous IGF-I, and the levels of the 150 kDa complex, a ternary complex of IGF-I with IGFPB-3 and an acid labile-subunit, were found to be lower in 4 weeks old rats than in 7 weeks old rats. In rats at 4 weeks of age, the elimination of rhIGF-I was significantly faster than for the 7 week old rats, which would be due to the lower plasma levels of IGFBP-3 in young growing rats. PMID- 9344225 TI - High responsiveness of cytosolic free calcium concentration to angiotensin II in cultured pulmonary arterial myocytes from pulmonary hypertensive rats. AB - Pulmonary arterial myocytes were cultured from normotensive and pulmonary hypertensive rats. Microfluorimetry of Ca2+ signals in fluo-3-loaded single myocytes at day 7 of culture was performed by a laser-scanned confocal imaging system. The resting level of cytoplasmic Ca2+ concentration ([Ca2+]i) in vascular myocytes obtained from hypertensive rats was higher than that in cultured myocytes obtained from normotensive rats. Angiotensin II elevated [Ca2+]i in the vascular myocytes cultured from both normotensive and hypertensive rats. However, a rise of [Ca2+]i induced by angiotensin II in the vascular myocytes obtained from pulmonary hypertensive rats was higher than that obtained from normotensive rats. On the other hand, the response of [Ca2+]i to A23187 did not differ between the vascular myocytes cultured from normotensive and hypertensive rats. The present results suggest that the resting and angiotensin II-responsive levels of [Ca2+]i in pulmonary arterial myocytes cultured from pulmonary hypertensive rats are higher than those cultured from normotensive rats. PMID- 9344226 TI - Hepatic uptake of p-nitrophenyl sulfate by transporter that acetaminophen sulfate shares for uptake: sulfate moiety as a vector for metabolite transport. AB - Hepatic uptake of the sulfate conjugate of p-nitrophenol (p-NPsul) has been studied. Uptake clearance of p-NPsul by isolated rat hepatocytes was dependent on p-NPsul concentration, suggesting carrier-mediated transport. The uptake of p NPsul by isolated rat hepatocytes was inhibited by acetaminophen sulfate (APAPsul), the uptake of which had been previously reported to be inhibited by p NPsul. The hepatic uptake of p-NPsul was also inhibited by bromosulfophthalein (BSP) or dibromosulfophthalein (DBSP), which had been reported to inhibit hepatic uptake of APAPsul in the previous study. These inhibitory effects were observed in isolated perfused liver experiments as well as in isolated hepatocyte experiments. Therefore, it was concluded that hepatic uptake of p-NPsul was mediated by a transporter, which mediates hepatic uptake of APAPsul as well as BSP or DBSP. It is suggested that sulfate moiety may have a key role as a vector for the hepatic transport of sulfate conjugate metabolites. Interaction between sulfate conjugate metabolites in hepatic uptake may also be expected. PMID- 9344227 TI - Association between hyperlipidemia and hepatic cytochrome P-450 in guinea pigs. AB - The effects of ascorbic acid (AA) or vitamin C in atherosclerosis has attracted considerable attention; however results of clinical studies are conflicting. Several studies indicate an increase in plasma triglyceride (TG) and cholesterol (CH) levels in guinea pigs (GP) that have been fed a diet containing a minimal amount of AA. Previous studies carried out in GP fed a diet devoid of AA showed a significant decrease in cytochrome P-450 level compared to GP fed high and adequate amounts; however, the level of cytochrome P-450 in the two groups were not significantly different. The enzymes that synthesize TG and CH are located in endoplasmic reticulum which is also the site for cytochrome P-450 synthesis. It is of interest to determine whether there is an association between TG and CH synthesis and cytochrome P-450 induction. Adult male Hartley GP weighing 350-400 g were fed a diet containing 2.5% (Group I), 0.1% (Group II) and 0% (Group III) AA. The food consumption and weight gain were not significantly different in different groups. After feeding the diet for four weeks, half of the animals in each group were starved. Blood was withdrawn and TG and CH were determined in the serum. TG and CH were markedly elevated in both starved and nonstarved Group III GP; however, these levels were not altered in Group 1 and Group II GP. Plasma AA showed significant differences in all three nonstarved and starved groups. Plasma alpha-lipoprotein was decreased and beta-lipoprotein was increased in Group III GP. Hepatic CH and TG were also significantly elevated in Group III GP, and Groups I and II showed no changes. TG and CH showed a negative correlation with cytochrome P-450, whereas CH and TG showed a positive correlation. We conclude that AA deficiency causes extensive hyperlipidemia, feeding high level of AA does not alter the lipid metabolism and induction ofcytochrome P-450 is inversely related to TG and CH synthesis. PMID- 9344228 TI - Preliminary evaluation of the anticonvulsant activity of a valproic acid analog: N-(2-propylpentanoyl) urea. AB - Anticonvulsant activity, lethality and neurotoxicity of a valproic acid (VPA) analog, N-(2-propylpentanoyl) urea (VPU) in comparison to its parent compound were investigated in mice. Intraperitoneally administered VPU demonstrated a higher protection than VPA in both the maximal electroshock seizure (MES) and the pentylenetetrazole (PTZ) tests exhibiting a median effective dose (ED50) of 66 and 57 mg/kg, respectively. VPU weakly blocked the effect of bicuculline and was ineffective in strychnine test. Furthermore, VPU was also active orally demonstrating an ED50 approximately 6 times higher than its ED50 by the intraperitoneal route. Based on the relatively high median lethal dose (LD50), 1553 mg/kg, VPU possesses a greater margin of safety (LD50/ED50) than did VPA. Unwanted (side) effects in terms of impairment of motor activity and neurotoxicity were assessed by the rotorod test, locomotor activity test as well as potentiation of barbiturate sleeping time. The median neurotoxic dose (TD50) as measured by rotorod test were 625 mg/kg for intraperitoneally given VPU. This finding results in higher protective index (PI = TD50/ED50) of VPU (PI = 9.5) than that of VPA (PI = 1.1) implying that, in therapeutic dose, VPU may produce less neurological side effects than did VPA. Superiority of VPU in terms of higher potency in parallel with minimal neurological deficit as assessed by rotorod test was evident throughout the observation period of 12 hours. Similar results on locomotor activity as well as potentiation of barbiturate sleeping time were obtained with VPU and VPA. Thus, VPU is preferably expected to exert minor degree of CNS depression. Taken altogether, our findings demonstrate greater anticovulsant activity for VPU than for VPA. In addition, this compound is also orally active and seems to offer a greater safety margin in parallel with lower unwanted effects in relation to its parent compound. As indicated by the animal data obtained, VPU is an attractive anticonvulsant candidate for further investigation. PMID- 9344229 TI - Determination of a new antiulcer agent, eupatilin, in rat plasma, bile, urine, and liver homogenate by high-performance liquid chromatography. AB - A high-performance liquid chromatographic method was developed for the determination of a new antiulcer agent, eupatilin, in rat plasma, urine, bile, and liver homogenate. The method involved deproteinization of biological sample with the same volume of acetonitrile. A 100 microl aliquot of the supernatant was injected onto a C18 reversed-phase column. The mobile phase employed was ammonium acetate buffer (1% ammonium acetate and 0.5% acetic acid) - acetonitrile (58:42, v/v) and the flow rate was 1.0 ml/min. The column effluent was monitored by a ultraviolet detector set at 350 nm. The retention time for eupatilin was approximately 6.5 min. The detection limits for eupatilin in rat plasma, urine, bile, and liver homogenate were 50, 50, 100, and 100 ng/ml, respectively. The coefficients of variation of the assay were generally low (below 7.46%) for rat plasma, urine, bile, and liver homogenate. No interferences from endogenous substances were observed. PMID- 9344230 TI - Gastric anti-ulcer and cytoprotective effect of l-serine in rats. AB - Effect of l-serine has been studied for its ability to inhibit gastric secretion and to protect the gastric mucosa against stress and chemically induced ulcers. Acid secretion studies were undertaken in pylorus-ligated rats with and without l serine treatment. Experimental gastric lesions were induced by hypothermic restraint stress, indomethacin and necrotizing agents including 80% ethanol, 0.2 M sodium hydroxide and 0.6 M hydrochloric acid in rats. The level of nonprotein sulfhydryl compounds and gastric wall mucus were also measured in the glandular stomach of the rats following ethanol-induced gastric lesions. The results of this study demonstrate that l-serine produces a dose-dependent inhibition of gastric acid secretions in rats. Pretreatment with l-serine also attenuated the formation of stress-, indomethacin- and necrotizing agents-induced gastric lesions. The antiulcer activity of l-serine was associated with significant inhibition of ethanol-induced depletion of nonprotein sulfhydryls and gastric wall mucus. In conclusion, this study demonstrates that l-serine possesses significant antiulcer and cytoprotective activity against various experimentally induced gastric lesions. Although the mechanism of action of l-serine requires further evaluation, the experimental observations derived from this study may have future clinical relevance and therefore deserve to be investigated thoroughly. PMID- 9344231 TI - Studies on the antisecretory, gastric anti-ulcer and cytoprotective properties of glycine. AB - Glycine, a neutral amino acid has been studied for its ability to inhibit gastric secretion and to protect the gastric mucosa against chemically and stress-induced ulcers. Acid secretion studies were undertaken in pylorus-ligated rats with and without glycine treatment. Experimental gastric lesions were induced by hypothermic-restraint stress, indomethacin and necrotizing agents including 80% ethanol, 0.2 M sodium hydroxide and 0.6 M hydrochloric acid in rats. The level of nonprotein sulfhydryl compounds and gastric wall mucus were also measured in the glandular stomach of the rats following ethanol-induced gastric lesions. The results of this study demonstrate that glycine dose dependently reduced the gastric secretions in rats. Pretreatment with glycine significantly protected animals against stress-, indomethacin- and necrotizing agents induced gastric lesions. The antiulcer activity of glycine was associated with significant inhibition of ethanol-induced depletion of nonprotein sulfhydryls and gastric wall mucus. In conclusion, this study demonstrates that glycine possesses significant antiulcer and cytoprotective activity. However, further detailed studies are warranted to establish the mechanism(s) of action, and to determine its role in the prophylaxis and treatment of gastric ulcer disease. PMID- 9344232 TI - Protection by glycine against hypoxia-reoxygenation induced hepatic injury. AB - Isolated perfused livers from rats fasted 16 h before surgery showed a strong decrease in oxygen consumption as well as hepatotoxic responses when subjected to 30 min of hypoxia (95%, N2/5% CO2) followed by 90 min of reoxygenation (95% O2/5% CO2). Toxicity was evident by a release of enzymes (LDH, GPT, GLDH) into the perfusate and by a nearly complete suppression of bile flow. Hepatic reduced gluthathione dropped to about 20% and hepatic ATP to about 50% of the initial values. Furthermore, the concentrations of thiobarbituric acid reactive (TBA) material increased eightfold in the perfusate and by 70% of the control values in the livers. Glycine added to the perfusate at concentrations of 3, 6 and 12 mmol/l prevented dose-dependently all measures of hepatotoxicity as well as the indices of lipid peroxidation induced by hypoxia/reoxygenation. A maximal and nearly complete protection was obtained with 12 mmol/l glycine. Glycine increased the bile flow of perfused livers not subjected to hypoxia and attenuated the drop of bile flow induced by hypoxia-reoxygenation. Ligation of the bile duct, however, did not influence the cytoprotective effects of glycine in hypoxia reoxygenation induced hepatic injury. In conclusion, glycine is an effective antidote against hypoxia-regoxygenation induced injury of the isolated rat liver. PMID- 9344233 TI - Induction of differentiation of U-937 cells by 2-chloro-3-amino-1,4 naphthoquinone. AB - Naphthoquinone compounds have various pharmacological effects such as antiviral, antifungal and anticancer activities. We demonstrated the differentiation of the inducing effect of a naphthoquinone derivative, 2-chloro-3-amino-1,4 nahpthoquinone (NQCA) on the human leukemia cell line U-937. When U-937 cells were treated with NQCA for 4 days, phenotypes indicative of differentiation such as nitroblue tetrazolium (NBT)-reducing activity and phagocytosis were induced. To evaluate the route of differentiation of U-937 cells induced by NQCA, we determined naphthol AS-D chloroacetate esterase and alpha-naphthyl acetate esterase activities. Four days treatment of U-937 cells with NQCA increased alpha naphthyl acetate esterase activity about 63.5% but naphthol AS-D chloroacetate esterase was not detected. These results indicate that NQCA caused differentiation of U-937 cells into macrophage-like cells. Since protein kinase C (PKC) and protein kinase A (PKA) have important roles in cell-differentiation and proliferation, we employed a PKC inhibitor NA-382 and a PKA inhibitor H-89 to examine the effects of each kinase on the differentiation of U-937 cells. The PKC inhibitor NA-382 decreased the effect of NQCA on U-937 cells, while the PKA inhibitor H-89 did not. Also glutathione (GSH) inhibited the effect of NQCA. It is concluded that the differentiation-inducing effect of NQCA on U-937 cells may be attributed to PKC activation followed by production of free radicals. PMID- 9344234 TI - Neutrophil elastase inhibitor (ONO-5046-Na) inhibits the growth of human lung cancer cell lines transplanted into severe combined immunodeficiency (scid) mice. AB - We examined the in vivo effects of ONO-5046xNa, a specific neutrophil elastase (NE) inhibitor, on the growth of 2 non-small cell lung cancer cell lines, EBC-1 and PC-3, transplanted into severe combined immunodeficiency (scid) mice. The daily intraperitoneal injection of ONO-5046xNa (50 mg/kg/day) completely suppressed the tumor growth in EBC-1, a human squamous carcinoma cell line which produces immunoreactive NE. By contrast, in PC-3, a human adenocarcinoma cell line which is unable to produce immunoreactive NE, the ONO-5046xNa treatment caused delayed growth of the tumor. These findings suggest that ONO-5046xNa may have a clinical role in preventing the growth of human non-small cell lung cancer. PMID- 9344235 TI - Cadmium and/or selenium effects on guinea pig lung PGE2, TXB2 and LTC4. AB - Male Hartley guinea pigs (480-610 g; n=5) were treated intratracheally with saline, cadmium (Cd, 0.3 mg) as cadmium chloride, selenium (Se, 0.3 or 0.06 mg) as sodium selenite or Se (0.06 mg) and Cd (0.3 mg). After 24 hours, lungs were collected and analyzed for prostaglandin (PGE2), thromboxane (TXB2) and leukotriene (LTC4) levels. Results indicated that, 0.3 mg Se and 0.06 mg Se in combination with 0.3 mg Cd increased PGE2 significantly. Selenium and Cd alone or in combination, decreased LTC4 and TXB2 significantly. PMID- 9344236 TI - Optimized differential display and reamplification parameters for silver staining. AB - Differential display (DD) is a powerful instrument to detect differences in gene expression of malignant and normal cells. An optimized silver staining protocol was used to compare mRNA expression of keratinocytes and squamous cell carcinoma cells of the head and neck. Optimal concentration for upstream and downstream primer was 0.2 microM. Best primer concentrations for reamplification were between 40 and 60 pM. With this optimized protocol nearly 50 differentially expressed transcripts have been identified and differential display can be applied safer, easier, and faster, than by radioactive labeling. PMID- 9344237 TI - Role of phosphoinositide-3-OH kinase in Ras signaling. PMID- 9344238 TI - From phosphorylase to phosphorylase kinase. PMID- 9344239 TI - Intrasteric regulation of protein kinases. PMID- 9344240 TI - Specificity in protein-tyrosine kinase signaling. PMID- 9344241 TI - Historical perspectives and new insights involving the MAP kinase cascades. PMID- 9344242 TI - Coupling transcription to signaling pathways: cAMP and nuclear factor cAMP responsive element modulator. PMID- 9344243 TI - Cyclic nucleotide-gated channels and calcium: an intimate relation. PMID- 9344244 TI - Chemoattractant receptor signaling: G protein-dependent and -independent pathways. PMID- 9344245 TI - Mitochondrial alpha-ketoacid dehydrogenase kinases: a new family of protein kinases. AB - Four mitochondrial protein kinases have been cloned. These proteins represent a new family of protein kinases, related by sequence to the bacterial protein kinases but by function to the eukaryotic serine protein kinases. Arg288 is required for recognition by BCKDK of the phosphorylation site on the E1alpha subunit of the BCKDH complex. BCKDK inhibits the dehydrogenase activity of the BCKDH complex by introducing a negative charge into the active-site pocket of the E1 component. Protein starvation of rats induces an increase in the amount of BCKDK bound to the BCKDH complex. This causes inactivation of the BCKDH complex and conserves branched-chain amino acids for protein synthesis in the protein starved state. Expression of the different PDK isoenzymes is tissue specific, and the different PDK isoenzymes are unique with respect to kinetic parameters for ATP and ADP and sensitivity to allosteric effectors (NADH, NAD+, coenzyme A, acetyl-CoA, pyruvate, and dichloroacetate). Preliminary experiments indicate that an increased amount of PDK2 protein partly explains the increase in PDK activity that occurs in rat liver in response to chemically induced diabetes. PMID- 9344246 TI - Phosphatases as partners in signaling networks. PMID- 9344247 TI - The cAMP in thyroid: from the TSH receptor to mitogenesis and tumorigenesis. PMID- 9344248 TI - Ca2+/calmodulin-dependent myosin light-chain kinases. PMID- 9344249 TI - Cascade activation of the calmodulin kinase family. PMID- 9344250 TI - Modulation of sodium and calcium channels by protein phosphorylation and G proteins. PMID- 9344251 TI - Interruption of specific guanylyl cyclase signaling pathways. PMID- 9344252 TI - Structure, function, and regulation of human cAMP-dependent protein kinases. AB - A large number of hormones, neurotransmitters, and other signaling substances that bind to G-protein-coupled cell-surface receptors have their signals converge at one sole second messenger, cAMP. The question of how specificity can be maintained in a signal-transduction system in which many extracellular signals leading to a vast array of intracellular responses are all mediated through one second-messenger system has been the subject of thorough investigation and a great deal of speculation. An increasing number of cAK isozymes, consisting of homo- or heterodimers of R subunits (RIalpha, RIbeta, RIIalpha, RIIbeta) with associated catalytic subunits (C alpha, Cbeta, Cgamma), may, at least in part, explain this specificity. The various cAK isozymes display distinct biochemical properties, and the heterogeneous subunits of cAK reveal cell-specific expression and differential regulation at the level of gene transcription, mRNA stability, and protein stability in response to a wide range of hormones and other signaling substances. The existence of a number of anchoring proteins specific to either RIIalpha or RIIbeta, and which localize cAKII isozymes toward distinct substrates at defined subcellular loci, strongly supports the idea that specific functions can be assigned to the various cAK isozymes. The demonstration that selective activation of cAKI is necessary and sufficient for cAMP-mediated inhibition of T cell proliferation, and the observation that T-cell activation is associated with redistribution and colocalization of cAKI to the TCR, is also compatible with the notion of isozyme-specific effects. PMID- 9344253 TI - Structural order of the slow and fast intrasubunit cGMP-binding sites of type I alpha cGMP-dependent protein kinase. PMID- 9344254 TI - The pseudosubstrate sequences alone are not sufficient for potent autoinhibition of cAMP- and cGMP-dependent protein kinases as determined by synthetic peptide analysis. PMID- 9344255 TI - Recent advances in the study of Ca2+/CaM-activated phosphodiesterases: expression and physiological functions. PMID- 9344256 TI - Specificity and complexity of receptor-G-protein interaction. PMID- 9344257 TI - G-protein-coupled receptors and their regulation: activation of the MAP kinase signaling pathway by G-protein-coupled receptors. AB - G-protein-coupled receptors that mediate cellular responses to a variety of humoral, endothelial-, or platelet-derived substances are able to stimulate MAP kinase activity. In transfected model systems, G-protein-coupled receptors that couple to pertussis toxin-insensitive G proteins of the Gq/11 family mediate this activation predominantly via a PKC-dependent mechanism. In contrast, activation of MAP kinase by receptors that couple to pertussis toxin-sensitive Gi proteins is PKC-independent and requires downstream activation of the low-molecular-weight G protein, Ras. This pathway can be inhibited by coexpression of peptides that sequester Gbetagamma subunits, and is mimicked by overexpression of Gbetagamma subunits. This Ras-dependent MAP kinase activation requires tyrosine phosphorylation of "docking proteins," including the shc adapter protein, and depends upon recruitment of Grb2/Sos1 complexes to the plasma membrane, thus resembling the pathway of MAP kinase activation employed by the receptor tyrosine kinases. Other molecules, including PI-3-kinases and phosphotyrosine phosphatases, probably also contribute to Gbetagamma-subunit-mediated assembly of a mitogenic signaling complex. Identification of the G-protein-coupled, receptor regulated tyrosine kinase(s), and the means by which the mitogenic signaling complex is assembled at the plasma membrane, remain subjects of further study. PMID- 9344258 TI - Rab3A-rabphilin-3A system in neurotransmitter release. PMID- 9344259 TI - The process of amputation and rehabilitation. AB - The process of rehabilitation of the amputee begins before any surgical intervention. It embodies a team concept with the patient at the core. Evaluation of the appropriate amputation level involves a number of parameters and input from all the team members. Goals are modified as the rehabilitation progresses. The involvement of family and/or significant other is crucial for a positive outcome. Whether or not the patient is a prosthetic candidate, the rehabilitation process is designed to achieve maximum independence, and an improved quality of life. PMID- 9344260 TI - The metabolic cost of lower extremity amputation. AB - Amputation surgery should be approached as the first step in the rehabilitation of a patient with a non-functioning, salvageable limb. Before performing amputation surgery, the rehabilitation team should have an understanding of outcome expectations for the individual patient. Biologic joints are energy couples. When performing amputation surgery, more proximal amputations, accompanied by the removal of more joints, decreases the ability of patients to walk and live independently. Functional outcome appears to increase with the length of the residual limb. PMID- 9344261 TI - Major foot trauma: the dilemma of reconstruction versus amputation. AB - Advances in medicine and surgery have allowed for dramatic improvements in the management of severe foot trauma. Some salvage attempts have led patients to repeated hospitalizations, extensive complications, and a poor functional outcome. The decision for an early amputation versus salvage of the foot remains a difficult one. Two case studies from Loyola University Medical Center and the current predictive indices for limb salvage are reviewed in this chapter. PMID- 9344262 TI - Tumor as an indication for amputation in the foot. AB - When a benign or malignant tumor presents itself in the foot, amputation may, at times, be necessary for complete eradication. Suspected malignancies require detailed general evaluation and oncology consultation. Biopsy is the gold standard for diagnosis. Depending on the type of tumor identified, resection margins of varying degrees may be necessary. Cases are used to illustrate the principles of tumor care. PMID- 9344263 TI - Pediatric lower extremity amputations. AB - Amputation in children is never undertaken without a great deal of thought. Even in less than optimum circumstances, salvage may be attempted because the patient is a child. This article reviews some of the congenital and acquired disorders that may be treated by amputation. The overlap of syndromes and the medical morbidities are also discussed. PMID- 9344264 TI - Amputation for infection or ischemia. AB - Successful limb salvage for infection or ischemia is dependent on the surgeon's ability to stabilize the acute process and select a level of amputation that will result in predictable healing potential and functional longevity. These patients often have multiple medical problems and significant social issues that must be considered in their overall care. This article presents current concepts in limb salvage surgery and reviews wound healing parameters to assist the surgeon in selecting the appropriate level of amputation for these patients. PMID- 9344265 TI - Toe amputations and ray resections. AB - Loss of a part of the lower extremity is an unfortunate complication of diabetes. Indications and general principles of amputation have been established. Distal limb salvage procedures include forefoot amputation alternatives, digital amputations, and ray resections. A variety of risks and complications are associated with these procedures. Postoperative management including prosthetic and accommodative therapy may enhance the successful outcomes of these procedures. PMID- 9344266 TI - Transmetatarsal and midfoot amputations. AB - This article discusses transmetatarsal and midfoot amputations, selection of level, criteria for wound healing, surgical techniques, and functional considerations. Amputations through the middle of the foot include the Lisfranc amputation at the tarsometatarsal joints and the Chopart amputation at the midtarsal joints. Both of these procedures result in the development of equinovarus deformity, and require lengthening of the Achilles tendon. Transmetatarsal amputation preserves foot function, is cosmetically acceptable and does not require a prosthesis. Satisfactory limb salvage can be achieved by the motivated patient and knowledgeable surgeon when these procedures are employed. PMID- 9344267 TI - Syme's ankle disarticulation: the transmalleolar amputation. AB - Salvage of the infected extremity in diabetic patients continues to be a complex problem. Amputation at the Syme's level helps maintain patient independence and decreases the rehabilitation expense associated with more proximal amputation. Although the wound complication rates are relatively high, the ultimate healing rate and favorable functional outcomes in this high risk patient population make this an outstanding amputation level for pedal amputation. PMID- 9344268 TI - Prostheses, orthoses, and shoes for partial foot amputees. AB - A variety of devices are available for aftercare of the partially amputated foot. The prescription of these devices depends on the extent of the amputation, subsequent muscle balance, and the willingness of the patient to comply with the use of the device. Above all, long-term follow-up care is necessary to evaluate the need and effectiveness of any orthotic prescription as well as to provide local care for excessive preulcerative keratosis associated with new focal pressure points. PMID- 9344269 TI - Long-term aftercare and prevention of further amputation. AB - This article discusses the recently instituted PACT (Prevention Amputation Care and Treatment) program now being used at the Cleveland Veterans Affairs Medical Center. This program is a multidisciplinary interventional prevention strategy that includes both physical and psychosocial components, addressing the long-term after-care for patients postamputation and preventive interventions for those at risk. PMID- 9344270 TI - The unique oral health needs of an aging population. AB - Over the last century, the number and percentage of older adults has increased dramatically. In the last 30 years, the percentage of older edentulous adults has declined significantly but the total number is expected to remain constant at 9 million until the year 2020. The increasing number of and percentage of dentate adults will have more teeth at risk for caries and periodontal disease. Many of these adults will have multiple medical problems and be taking various pharmacotherapies which will complicate oral disease and its treatment. New concepts in prevention of oral disease will be required, as will more accurate diagnostic procedures, especially to identify the at-risk older adults. PMID- 9344271 TI - Physiologic changes in the elderly. AB - A variety of age-related changes in the oral cavity and throughout the aging body can affect dental care and treatment plans. Some of these changes may be unavoidable features of senescence. Others, previously thought to be part of "normal aging," may be modifiable with lifestyle choices or may represent subclinical pathological processes. The ability to compensate for losses in system capacity varies among individuals and may result in functional changes ranging from substantial to unmeasurable. An appreciation of the intricacy of these relationships, and coordinated treatment with the primary care physician, can enhance the dental care of the aged patient. PMID- 9344272 TI - Medical history and risk assessment. AB - The "graying" of America has resulted in dentists treating increased numbers of elderly patients, 60% of whom are dentate. Since the majority of elderly persons has at least one chronic disease, this chapter addresses critical aspects of history taking and risk assessment for the geriatric dental patient. Self administered questionnaires have limitations in the geriatric population and the medical history must emphasize functional status, medication use, social support, and financial considerations. Common chronic diseases which potentially increase the risk of adverse events for the geriatric patient undergoing dental care are discussed. Effective dentist-physician communication is pivotal to the successful management of the elderly dental patient. PMID- 9344273 TI - Nutrition and oral health in the elderly. AB - Nutritional status of the aging patient is inextricably linked to his or her oral health. Various screening instruments make it easier for the dentist to determine the nutritional status of the patient. This article discusses several of those instruments, in addition to further consideration of the link between nutrition and oral health in the aging patient. PMID- 9344274 TI - The most frequently prescribed medications in the elderly and their impact on dental treatment. AB - The high prevalence of multiple chronic medical conditions in the elderly and the likelihood of multiple drug therapies dramatically increase the chance of drug specific adverse effects and drug-drug interactions. This article reviews the age associated alterations in pharmacokinetics and pharmacodynamics along with the most frequently prescribed medications for the elderly. Adverse drug reactions, drug-drug interactions and oral side effects are reviewed along with the impact these medications may have on dental treatment planning and management. PMID- 9344275 TI - Oral health promotion and prevention for older adults. AB - An oral health promotion and prevention program customized to individual needs begins with a thorough assessment of function and risk profile for dental diseases. Toothbrushes and interproximal cleaners can be selected or adapted to meet special needs of older adults. Fluoride use based on caries risk is an important adjunct to any prevention program. Other preventive agents such as chlorhexidine rinses and xylitol gum supplement the program as risk factors increase or when health and disability limit the ability to effectively perform oral hygiene procedures. Oral cancer screening examination is advocated on a regular basis for all older persons. PMID- 9344276 TI - Periodontal care. AB - Prevalence of periodontal disease increases with increasing age. Recent surveys show a decline in edentulism, resulting in an increase in the number of teeth at risk for moderate periodontal disease, with more severe disease restricted to a small portion of the population. Most evidence indicates that age associated increases in disease prevalence and severity is not caused by the process of aging, but is related to past disease history, social, and behavioral factors. Chronologic age is not a contradiction to any modality of periodontal treatment. Diagnosis and treatment of older patients may be modified by the increasing prevalence of chronic diseases, and by concomitant use of multiple medications. PMID- 9344277 TI - Root caries in the older patient: significance, prevention, and treatment. AB - Root caries is an emerging challenge to the dental professions because of the growing number of increasingly aging adults who have retained many or all of their teeth. Risk factors for developing root caries point to both intraoral and environmental factors, making the management of root caries complex and multidisciplinary. Prevention based on a composite of risk factors is the most desirable approach for management. Patients who have developed caries of the roots can be treated with remineralization strategies, recontouring techniques, intracoronal restorations of a variety of established and recently introduced materials, or extracoronal restoration. Dental professionals need to keep abreast of new approaches that are emerging for the management of root caries. PMID- 9344278 TI - Endodontic considerations in the geriatric patient. AB - Endodontic needs of today's and tomorrow's growing older adult population present increasing challenges for dental care providers. Biologic and anatomic differences in the dental tissues between older and younger patients must be understood and considered in treatment planning and performance for appropriate endodontic procedures. These differences generally do not contraindicate treatment, which, when performed correctly, will be successful in the elderly patient. PMID- 9344279 TI - Prosthodontic considerations for the older patient. AB - The elderly have both the greatest level of need for prosthodontic services of any age group, and the greatest degree of complicating dental, medical, and behavioral factors. Issues arise in daily practice of whether or not to replace a missing tooth or teeth for a patient of advanced age and a wide variety of challenges-dental/oral and others-face the dentist who is considering replacing some or all of an older person's teeth. This article focuses on clinical approaches and techniques that have proven particularly important and useful for providing prosthodontic care to the older adult. PMID- 9344280 TI - Treatment planning of the elderly implant patient. AB - The prevalence of partial and complete edentulism in the older adult patient and the predictability of specific types of dental implants obligates the dental health professional to consider the fabrication of implant-related prostheses as an alternative treatment option. Special emphasis is placed in this article on the provision of clip-bat overdentures in the treatment of the fully edentulous patient and their relative advantages compared to a fixed-hybrid prosthesis. PMID- 9344281 TI - Management of dry mouth. AB - Dry mouth is a common complaint and, when associated with salivary gland dysfunction, a significant problem. Accurate and complete diagnosis of the patient with complaints of xerostomia is essential. Management should be directed to relief of symptoms, control of oral disease, and improvement in salivary function. With a systematic approach and aggressive management, most dry mouth patients can achieve oral comfort and adequate oral function. PMID- 9344282 TI - Diagnosis and treatment of common oral lesions found in the elderly. AB - A wide variety of oral lesions are recognized in the geriatric patient. The most common lesions include neoplasia, immunologic based mucosal disease, hematological disorders, oral manifestation of systemic disease, and conditions characterized by oral or facial pain. Diagnostic and treatment considerations for leukoplakia, carcinoma, metastatic disease, candidiasis, herpes zoster, plasmacytoma, myeloma, lymphoma, several of the more common vesiculoulcerative mucosal diseases and idiopathic burning mouth syndrome are presented. PMID- 9344283 TI - Diagnosis and patient management of oral cancer. AB - With an average age of 60 at diagnosis, oral cancer is largely a disease of older adults. This article reviews the incidence, risk factors, early detection and diagnosis of oral cancer, as well as the principles of multidisciplinary management. Considerations for dental treatment planning prior to radiation therapy and surgery for oral cancer are included. PMID- 9344284 TI - Providing dental care for patients diagnosed with Alzheimer's disease. AB - Alzheimer's disease is the most common dementing illness affecting over 4 million Americans. As the population ages, dentists and other health care providers will be faced with the daunting task of managing an increasing number of people with this disease. Currently, there are no definitive medications to treat this disease, although there are a number of recent drugs which may help to alleviate some symptoms. This article reviews the current medical treatment and the dental concerns which face the dentist, patient, and family. Suggestions for dental management are given along with practical recommendations for caregivers. PMID- 9344286 TI - Peer review and Academic Radiology. PMID- 9344285 TI - Fitting the pieces together: treatment planning in the geriatric dental patient. AB - Treatment planning older patients often becomes a complex process as dental professionals, patients, family, and caregivers attempt to prioritize and balance the influences of multiple age-associated dental, systemic, and psychosocial factors. To assist clinicians in identifying and weighing numerous factors that can influence planning dental care for older patients, clinicians should be wary of relying on chronological age as a factor, but should focus on the issues of biologic age and life expectancy, which may be greater than many older adults believe. The longevity of dental interventions is another factor that is helpful to consider in determining the most appropriate treatment plan for older adults. Among many issues influencing the treatment planning process, the quality of communication between clinicians and older patients is critical, along with the influence of third parties, including families and professional caregivers. Due to the lack of objective information on the outcomes of dental care in the older population, clinicians inevitably face many situations in which there is uncertainty about the best course of therapy. Practitioners can adopt specific strategies to help minimize difficulties that may be associated with the provision of care under such circumstances. PMID- 9344287 TI - Teaching as a science. PMID- 9344288 TI - Coronary artery calcium: alternate methods for accurate and reproducible quantitation. AB - RATIONALE AND OBJECTIVES: The aim of this study was to determine a more precise and accurate method of quantitating coronary artery calcium (CAC) detected with electron-beam computed tomography (CT) in patients with low CAC scores. MATERIALS AND METHODS: Two 40-section, 3-mm-collimation, electrocardiographically gated electron-beam CT examinations of the heart were performed in each patient. Fifty patients with average scores between 2 and 100, as determined with the conventional scoring algorithm, were selected. The modified conventional scoring algorithm was compared with two techniques: calculated calcium volume and approximated calcium mass. RESULTS: The percentage difference between scans ranged from 37.2% for the conventional scoring method to 28.2% and 28.4% for volume- and mass-based methods, respectively. Increasing lesion size thresholds does not improve quantitative precision and reduces accuracy in patients with small amounts of CAC. CONCLUSION: Quantification methods based on calcification volume or mass decrease score variation compared with the conventional scoring method, and increased size threshold does not improve accuracy. PMID- 9344289 TI - Evaluation of in vivo total and regional air content and distribution in primate lungs with high-resolution CT. AB - RATIONALE AND OBJECTIVES: The authors sought to determine whether gray-scale quantitative information from high-resolution computed tomography (CT) could reliably yield estimates of lung air content and help determine changes in air content with lung inflation and deflation. MATERIALS AND METHODS: High-resolution CT images (n = 40) of lungs of two anesthetized monkeys were obtained after inflation with known air volumes. Percentage air content was calculated for each voxel, and lung volumes and patterns of air distribution were determined. RESULTS: When corrected for pressure and temperature, high-resolution CT-based volumes correlated closely with inflation volumes (r = .99; standard error = 3.4%). Patterns of regional change in air content demonstrated known patterns of ventilation. CONCLUSION: Although the high-spatial-frequency algorithm used in high-resolution CT enhances edges of structures and improves visualization of anatomic detail, gray-scale values from the same high-resolution CT data set remain a reliable index of regional lung attenuation. PMID- 9344290 TI - Extent of myocardial collateralization: determination with three-dimensional elastic-subtraction spiral CT. AB - RATIONALE AND OBJECTIVES: This study was undertaken to develop a standard that can be used to assess new high-resolution collateral zone imaging methods. MATERIALS AND METHODS: The authors performed ex vivo helical CT in seven pig hearts after microsphere studies of blood flow and coronary angiography. They compared the zones of collateralization depicted at CT and at microsphere studies. RESULTS: The extent of the collateral zone at CT, computed by using elastic subtraction, correlated well with the coronary blood flow distribution determined with microsphere analysis (r = .95). The root-mean-square error was 6.5%, which indicates good agreement. CONCLUSION: Accurate assessment of collateralization extent has become an important goal because of the discovery of agents that stimulate the growth of coronary collateral vessels. The precision of elastic-subtraction CT and its validation with respect to the blood flow distribution at microsphere analysis indicate that elastic-subtraction CT can serve as a standard for the measurement of collateralization extent. PMID- 9344291 TI - Bronchogenic carcinoma: a survey of CT protocols for staging disease. AB - RATIONALE AND OBJECTIVES: To determine whether a standard computed tomographic (CT) protocol is used in the staging of lung cancer. MATERIALS AND METHODS: A questionnaire was designed to determine what type of CT scanner is used, whether intravenous contrast material is used, how often the abdomen is scanned and at what level, and the section thicknesses used in scanning the chest and abdomen in patients with lung cancer. A total of 1,118 survey forms were mailed to members of the Society of Thoracic Radiology and to all community hospitals in the United States with at least 300 beds. RESULTS: The authors received 520 responses (47%) to the 1,118 questionnaires mailed. Of these 520 responses, 140 were from society members, 256 were from hospitals with 300-500 beds, and 124 were from hospitals with more than 500 beds. One-half of hospital respondents used helical CT scanners. Significantly more society members used helical CT scanners (P < .001). Intravenous contrast material was used to opacify mediastinal blood vessels at 449 (86%) of 520 hospitals. Intravenous contrast material was used for liver scanning at 363 (82%) of 444 hospitals, but it was used less often at hospitals in the northeast region and by society members than at hospitals in other regions (P < .001). A mixture of section thicknesses was commonly used (252 [48%] of 520 responses) for scanning the chest; a thickness of 8-10 mm was used in scanning the abdomen at most hospitals (348 [78%] of 445 responses). CONCLUSION: No CT protocol is consistently used for the examination of patients with lung cancer. Use of intravenous contrast material during chest or liver CT also is not uniform. PMID- 9344293 TI - Fellowship in medical education research: an opportunity for academic radiologists. PMID- 9344292 TI - Three-dimensional vascular reconstruction with a clinical x-ray angiography system. AB - RATIONALE AND OBJECTIVES: The authors developed a technique to produce high resolution, three-dimensional images of vasculature from a set of x-ray projections in an attempt to provide detailed anatomic representations of complex vasculature. MATERIALS AND METHODS: Projection images were acquired with a clinical angiographic system by using biplanar rotational digital subtraction angiography. The images were reconstructed with an additive algebraic reconstruction technique. RESULTS: The feasibility of the technique was tested by reconstructing three-dimensional images of several phantoms, including a wire phantom and an anatomic flow phantom. The anatomic phantom allowed replication of contrast material flow and image noise that are characteristic of patient examinations. The reconstruction procedure was then used to examine a carotid artery and a cerebral aneurysm in two patients. CONCLUSION: A method of reconstructing vasculature from x-ray angiograms has been developed and validated with geometric and anatomic phantoms. Preliminary patient applications indicate that this technique enables enhanced visualization of complex vascular relationships and structures. PMID- 9344294 TI - Evaluation of a training program for persons with SCI paraplegia using the Parastep 1 ambulation system: part 1. Ambulation performance and anthropometric measures. AB - OBJECTIVE: To describe performance parameters and effects on anthropometric measures in spinal cord injured subjects training with the Parastep 1 system. DESIGN: Before-after trial. SETTING: Human spinal cord injury applied research laboratory. PARTICIPANTS: Thirteen men and 3 women with thoracic (T4-T11) motor complete spinal cord injury: mean age, 28.8yrs; mean duration postinjury, 3.8yrs. INTERVENTION: Thirty-two functional neuromuscular stimulation ambulation training sessions using a commercially available system (Parastep-1). The hybrid system consists of a microprocessor-controlled stimulator and a modified walking frame with finger-operated switches that permit the user to control the stimulation parameters and activate the stepping. OUTCOME MEASURES: Distance walked, time spent standing and walking, pace, circumferential (shoulders, chest, abdomen, waist, hips, upper arm, thigh, and calf) and skinfold (chest, triceps, axilla, subscapular, supraillium, abdomen, and thigh) measurements, body weight, thigh cross-sectional area, and calculated lean tissue. RESULTS: Statistically significant changes in distance, time standing and walking, and pace were found. Increases in thigh and calf girth, thigh cross-sectional area, and calculated lean tissue, as well as a decrease in thigh skinfold measure, were all statistically significant. CONCLUSIONS: The Parastep 1 system enables persons with thoracic-level spinal cord injuries to stand and ambulate short distances but with a high degree of performance variability across individuals. The factors that influence this variability have not been completely identified. PMID- 9344295 TI - Evaluation of a training program for persons with SCI paraplegia using the Parastep 1 ambulation system: part 2. Effects on physiological responses to peak arm ergometry. AB - OBJECTIVE: To examine the task-nonspecific effects of functional neuromuscular stimulation (FNS)-assisted ambulation training on the physiological responses of persons with paraplegia to upper extremity exercise challenge. DESIGN: Before after trial. SETTING: Human spinal cord injury (SCI) applied research laboratory. PARTICIPANTS: Twelve men and three women with motor- and sensory-complete thoracic-level SCI (T4-T11), mean age 28.2 +/- 6.8yrs (range, 21.1 to 45.2yrs), mean injury duration 3.7 +/- 3.0yrs (range, 7 to 8.8yrs). INTERVENTION: Thirty two sessions of FNS ambulation training using a commercial six-channel system (Parastep 1). This system is composed of a microprocessor-controlled electrical stimulation unit and a walking frame outfitted with finger switches that allow the user to independently control the system and stimulation parameters. OUTCOME MEASURES: Peak and subpeak physiological responses to arm ergometry testing and upper extremity strength measures, obtained before and after the FNS ambulation training. RESULTS: Statistically significant increases in peak values for time to fatigue, peak power output, and peak VO2 (all p < .001). Heart rate was significantly lower throughout subpeak levels of arm ergometry after the ambulation training (p < .05). Values of upper extremity strength were not significantly altered after training. CONCLUSIONS: FNS ambulation by persons with SCI paraplegia results in task-nonspecific training adaptations. Central cardiovascular adaptations were indicated as the primary source of these beneficial alterations in exercise responses. PMID- 9344296 TI - Evaluation of a training program for persons with SCI paraplegia using the Parastep 1 ambulation system: part 3. Lack of effect on bone mineral density. AB - OBJECTIVE: To determine if the bone mineral density loss seen after spinal cord injury (SCI) is reversed by a walking program using the Parastep 1 system. DESIGN: Before-after trial. SETTING: Human SCI applied research laboratory. PARTICIPANTS: Thirteen men and 3 women with thoracic motor- and sensory-complete SCI, mean age 28.8yrs, mean duration postinjury 3.8yrs. INTERVENTION: Thirty-two functional neuromuscular stimulation (FNS) ambulation training sessions using a commercially available system (Parastep 1). This system consists of a microprocessor-controlled stimulator and a modified walking frame with finger operated switches that permit the user to control the stimulation parameters and activate the stepping. OUTCOME MEASURE: Bone mineral density at the femoral head, neck, and Ward's triangle measured using a Lunar DP3 dual-photon densitometer. RESULTS: No significant change in bone mineral density was found using repeated measures analyses of variance. CONCLUSIONS: Axial loading combined with muscle stimulation and resistive exercise does not result in significant changes in bone mineral density in persons with complete paraplegia. PMID- 9344297 TI - Evaluation of a training program for persons with SCI paraplegia using the Parastep 1 ambulation system: part 4. Effect on physical self-concept and depression. AB - OBJECTIVE: To determine whether persons with spinal cord injury (SCI) paraplegia who participated in an electrical stimulation walking program experienced changes in measures of physical self-concept and depression. DESIGN: Before-after trial. SETTING: Human SCI applied research laboratory. PARTICIPANTS: Volunteer sample of 12 men and 3 women with SCI paraplegia, mean age 28.75 +/- 6.6yrs and mean duration of injury 3.8 +/- 3.2yrs. INTERVENTION: Thirty-two FNS ambulation training sessions using a commercially available system (Parastep 1). The hybrid system consists of a microprocessor-controlled stimulator and a modified walking frame with finger-operated switches that permit the user to control the stimulation parameters and activate the stepping. OUTCOME MEASURES: The Tennessee Self-Concept Scale (TSCS) and the Beck Depression Inventory (BDI) were administered before and after training. Only the Physical Self subscale of the TSCS was analyzed. After training, individual interviews were performed to assess participants' subjective reactions to the training program. RESULTS: A repeated measures analysis of variance indicated that desired directional and statistically significant changes occurred on the Physical Self subscale of the TSCS (F(1,14) = 8.54, p < .011) and on the BDI (F(1,14) = 5.42, p < .035). CONCLUSIONS: Subsequent to the ambulation training program there were statistically significant increases in physical self-concept scores and decreases in depression scores. PMID- 9344298 TI - Evaluation of a training program for persons with SCI paraplegia using the Parastep 1 ambulation system: part 5. Lower extremity blood flow and hyperemic responses to occlusion are augmented by ambulation training. AB - OBJECTIVE: To test whether 12 weeks of exercise conditioning using functional neuromuscular stimulation (FNS) ambulation alters the resting lower extremity blood flow and hyperemic responses to vascular occlusion in subjects with paraplegia, and to determine whether an association exists between limb flow and lower extremity fat-free mass. DESIGN: Pretest, posttest. SETTING: Academic medical center. PARTICIPANTS: Subjects with chronic neurologically complete paraplegia. INTERVENTION: Thirty-two sessions of microprocessor-controlled ambulation using electrically stimulated contractions of lower extremity muscles and a rolling walker. OUTCOME MEASURES: Subjects underwent quantitative Doppler ultrasound examination of the common femoral artery (CFA) before and after training. End-diastolic arterial images and arterial flow-velocity profiles obtained at rest and after 5 minutes of suprasystolic thigh occlusion were computer-digitized for analysis of heart rate (HR), CFA peak systolic velocity (PSV), CFA cross-sectional area (CSA), flow velocity integral (FVI), pulse volume (PV), and CFA (arterial) inflow volume (AIV). RESULTS: Significant effects of training on CSA (p < .0001), FVI (p < .05), computed PV (p < .001), and computed AIV (p < .01) were observed. Resting HR was lower following training (p < .05). The change for resting PSV approached but did not reach significance (p = .083). Analysis of postocclusion PV and AIV showed significant effects for conditioning status (p values < .01), postcompression time (p values < .0001), and their interaction (p values < .01). At 1 minute after occlusion, the posttraining AIV response was 78.2% greater in absolute magnitude and 17.4% more robust when expressed as a percentage change from its resting value than before training. Significant correlations were found between thigh fat free mass and both AIV and PV (p values < .05). CONCLUSION: Exercise training using FNS ambulation increases the resting lower extremity AIV in individuals with paraplegia and augments the hyperemic response to vascular occlusion. Improved posttraining blood flow is attributable both to vascular structural changes and upregulation of vascular flow control mechanisms. Limb mass is associated with the volume of arterial blood flow. PMID- 9344299 TI - Mortality after spinal cord injury: an 11-year prospective study. AB - OBJECTIVE: To identify the relative risk of mortality after spinal cord injury (SCI) as a function of level of psychosocial, vocational, and medical adjustment. DESIGN: A prospective design was used: data on life adjustment was obtained at one time (1985), with subsequent survival status ascertained 11 years later (1996). Logistic regression was used to identify the relative risk of mortality given the level of adjustment on a number of predictor variables. SETTING: All participants were selected from outpatient files of a Midwestern university hospital. PARTICIPANTS: A total of 345 participants with SCI completed study materials in 1985 (a 78% response rate), 330 of whom could be definitively classified in 1996 as either survivor or deceased. Of these 330 participants, 84% were alive in 1996 (n = 278) and the other 16% were deceased (n = 52). MAIN OUTCOME MEASUREMENTS: The Life Situation Questionnaire (LSQ) was used to measure nine primary predictors related to life adjustment after SCI, including employment status and eight predictor scales: Medical Instability, Adjustment, General Satisfaction, Emotional Distress, Dependency, and Poor Health. The LSQ was also used to generate data on 34 individual items that were used in exploratory predictive analyses. RESULTS: All but one of the 8 primary adjustment predictors from 1985 significantly predicted 1996 mortality status. Dependency and low overall satisfaction were among the most significant predictors of mortality. CONCLUSIONS: Overall quality of life is important to the longevity of people with SCI, and comprehensive rehabilitation programs are needed to promote a level of life adjustment that maximizes longevity after SCI. PMID- 9344300 TI - Spouses of spinal cord injury survivors: the added impact of caregiving. AB - OBJECTIVE: To better understand the needs of spouses who provide care to spinal cord injury (SCI) survivors, by comparing their self-perceptions and complaints with those of their partners with disabilities and with those of spouses who do not provide care. DESIGN: Survey, including demographics, health concerns questionnaire, and administration of the Center for Epidemiologic Studies Depression Scale (CES-D), the Perceived Stress Scale (PSS), the Life Satisfaction Index (LSI-Z), and the Quality of Life and Individual Needs Questionnaire. SETTING: Two British SCI treatment centers, serving a defined population-based catchment area. PARTICIPANTS: One hundred twenty-four spouses of a longitudinally followed sample of SCI survivors, all of whom had been injured 23 or more years when the study was conducted in 1993. OUTCOME MEASURES: Scores on the above standardized tests, and responses to survey questions. RESULTS: Spouses had more depressive affect (p < .001) than their partners with disabilities, as measured by the CES-D. On the PSS, they exhibited no significant differences. Compared with spouses who were not caregivers, the caregiving spouses reported more physical stress (p = .005), emotional stress (p = .011), burnout (p = .007), fatigue (p = .002), and anger and resentment (p = .029). On the CES-D, they had more symptoms of depressive affect (p = .004) and somatic depression (p = .005). CONCLUSIONS: Spouses of long-term SCI survivors who fulfill a caregiving role report more symptoms of stress and depression than their partners with disabilities and other spouses who are not caregivers. PMID- 9344301 TI - Validity of the functional independence measure for persons with traumatic brain injury. AB - OBJECTIVE: Replicate and extend studies of the construct validity of the Functional Independence Measure (FIM) for persons with traumatic brain injury (TBI). DESIGN: A cross-sectional study of admissions to acute rehabilitation evaluated 6 months to 5 years after discharge. SETTING: An inpatient brain injury rehabilitation unit in a large, academic medical center. SUBJECTS: Ninety-five patients with primary diagnosis of TBI stratified by time postdischarge. MAIN OUTCOME MEASURES: Prediction of (1) average daily minutes of assistance and (2) supervision required in comparison to the Sickness Impact Profile (SIP) and SF 36. RESULTS: The FIM was highly predictive of minutes of assistance (83% accuracy), supervision (82% accuracy), and the need for either type of assistance (78% accuracy). Prediction was only minimally improved by measures of neurobehavioral impairment. The accuracy of the FIM was superior to the SIP and SF-36. CONCLUSIONS: Results provided substantial support for the validity of the FIM as a measure of functional independence for persons with TBI. The importance of supervision as a type of assistance required after TBI was evident, with the FIM highly predictive of this need, as well. PMID- 9344302 TI - Risk of seizure recurrence after the first late posttraumatic seizure. AB - OBJECTIVE: To determine the incidence and risk factors for seizure recurrence after the onset of late posttraumatic seizures (ie, seizures occurring more than 7 days after injury). DESIGN: Longitudinal cohort design. SETTING: Level 1 trauma center. PATIENTS: Sixty-three moderately to severely head-injured adults who developed late posttraumatic seizures during the course of their participation in a randomized, placebo-controlled study of the effectiveness of prophylactic phenytoin (Dilantin) for prevention of posttraumatic seizures. MAIN OUTCOME MEASURES: Time from the first unprovoked late seizure to time of seizure recurrence. RESULTS: The cumulative incidence of recurrent late seizures was 86% by approximately 2 years. However, the frequency of recurrent seizures varied considerably across subjects: 52% experienced at least five late seizures, and 37% had 10 or more late seizures within 2 years of the first late seizure. The relative risk of recurrence was highest in patients with a history of acute subdural hematoma and prolonged coma (ie, longer than 7 days). CONCLUSIONS: When late seizures develop after severe head injury, the probability of recurrence is high, which suggests that patients be treated aggressively with anticonvulsant medication after a first unprovoked late seizure. PMID- 9344303 TI - Promoting recovery in chronic aphasia with an interactive technology. AB - OBJECTIVE: To assess chronic aphasic patients' responses to resumption of therapy using an innovative, computer-based treatment system. DESIGN: Patients were assessed pretreatment and posttreatment using standardized assessment tools. Pretreatment and posttreatment performance score means were computed and compared, with statistical significance of the differences established using a one-tailed, matched t test. SETTING: The work was conducted at (1) a Veterans Affairs medical center participating in treatment research and (2) a regional aphasia center delivering therapy services for reimbursement. PATIENTS: Chronic aphasic patients (n = 23) from 6 months to more than 15 years postonset were enrolled in the study. They included a wide range of types and severities of aphasia, and all had received traditional speech-language therapy services earlier. INTERVENTIONS: All patients were treated in 1-hour clinical sessions by speech-language pathologists using the designated computer-based treatment system. All but one of the patients had access to the computer-based treatment system at home for practice between clinical therapy sessions. MAIN OUTCOME MEASURES: The outcome measures used were (1) the Porch Index of Communicative Ability (PICA), (2) the Boston Naming Test (BNT), (3) the Western Aphasia Battery (WAB), and (4) the Boston Diagnostic Aphasia Examination (BDAE). RESULTS: The majority of patients improved significantly in multiple modalities as assessed by these instruments. CONCLUSIONS: Specific measures of language function can be broadly, positively, and significantly influenced by computer-based language therapy in chronic aphasia. PMID- 9344304 TI - Speed of finger tapping and goal attainment after unilateral cerebral vascular accident. AB - OBJECTIVE: To determine (1) if speed of finger tapping is bilaterally slow after an acute unilateral cerebral vascular accident (CVA) and (2) if speed of finger tapping and grip strength are related to achieving rehabilitation goals during the first few weeks after stroke. DESIGN: Prospective inception cohort study. STUDY SETTING: Medical center and neurological institute. PARTICIPANTS: Fifty-one patients with unilateral CVAs. MAIN OUTCOME MEASURES: Documentation of goal attainment at discharge and bilateral measures of speed of finger tapping and grip strength. RESULTS: Speed of finger tapping and grip strength were often bilaterally below normal limits after an acute unilateral CVA, with the contralateral hand most affected. Speed of finger tapping, but not grip strength, in the ipsilateral hand was associated with achieving rehabilitation goals. Speed of finger tapping in the contralateral hand as well as bilateral grip strength was not related to achievement of rehabilitation goals. CONCLUSIONS: Motor findings suggest that bilateral cerebral dysfunction may be common after an acute unilateral CVA. The speed of finger movement in the hand ipsilateral to the lesion may reflect the degree to which the so-called "unaffected" cerebral hemisphere has in fact maintained its functional integrity. As such, it may be a useful behavioral marker for predicting goal attainment during early stages of neurorehabilitation. PMID- 9344305 TI - Gait pattern alteration by functional sensory substitution in healthy subjects and in diabetic subjects with peripheral neuropathy. AB - OBJECTIVE: To evaluate the ability of diabetic and nondiabetic individuals to learn to use a lower extremity sensory substitution device to cue gait pattern changes. DESIGN: Case-control study. SETTING: Gait laboratory. PARTICIPANTS: Thirty diabetic persons and 20 age- and education-matched nondiabetic controls responded to advertisements for study participation. INTERVENTION: Participants walked on a treadmill at three speeds (1, 2, and 2.5mph) with auditory sensory feedback to cue ground contact greater than 80% duration of baseline. MAIN OUTCOME MEASUREMENTS: The variables measured included gait cycle (steps per minute) and number of times per minute that any step during a trial exceeded 80% duration of ground contacted compared with a measured baseline step length for each speed. RESULTS: Persons in both groups were able to rapidly and significantly alter their gait patterns in response to signals from the sensory substitution device, by changing their gait cycles (nondiabetic group, F(17,124) = 5.27, p < .001; diabetic group, F(5,172) = 3.45, p < .001). Post hoc analyses showed early gait cycle modification and error reduction among both groups. The nondiabetic group learned to use the device significantly more quickly than the diabetic group during the slow (1mph, t = 3.57, p < .001) and average (2mph, t = 2.97, p < .05) trials. By the fast (2.5mph) ambulation trial, both groups were performing equally, suggesting a rapid rate of adjustment to the device. No technical failures from gait trainer malfunction occurred during the study. CONCLUSIONS: Diabetic persons with neuropathy effectively used lower extremity sensory substitution, and the technology is now available to manufacture a durable, effective lower extremity sensory substitution system. PMID- 9344306 TI - Frozen shoulder: correlation between the response to physical therapy and follow up shoulder arthrography. AB - OBJECTIVE: To study the correlation between improvement of shoulder motion and shoulder joint space capacity determinated by arthrography. DESIGN: Case series. SETTING: General community hospital. PATIENTS: Twelve patients with clinically diagnosed frozen shoulder without rotator cuff tear. All subjects were divided as "primary" and "secondary" according to spontaneous onset or not, and "acute" or "chronic" depending on whether duration of disease was less than 2 months or longer. INTERVENTIONS: Outpatient rehabilitation programs, including physical modalities, exercise intervention, and regular weekly outpatient clinic follow up. MAIN OUTCOME MEASURES: Shoulder range of motion (ROM) and joint space capacity in shoulder arthrography. RESULTS: In acute patients, the joint space capacity increased significantly after treatment (t = 2.82; p < .05). Increased joint space capacity was most significantly correlated with improvement in external rotation (r = .77, p < .05), followed by abduction (r = .43, p > .05), but was poorly correlated with flexion and internal rotation. In chronic patients, both primary and secondary groups, there was no obvious joint space capacity increase despite significant shoulder motion improvement. Follow-up arthrograms showed the reappearance and/or enlargement of the axillary recess and smoother capular margins in all the patients except one chronic case (disease duration for 1 year). These findings were more obvious in acute than in chronic patients. CONCLUSIONS: For frozen shoulder, generally described as "adhesive capsulitis," the adhesion was reversible in the acute stage. The increase of joint space capacity was significant and was correlated with improvement of external rotation. In chronic patients, ROM restoration occurred independent of change in joint space capacity, which increased slightly. The stretching of other contracted soft tissues around the shoulder, in addition to the adhesive capsule, may contribute to the recovery of chronic frozen shoulder. PMID- 9344307 TI - Cumulative trauma disorders in the upper extremities: reliability of the postural and repetitive risk-factors index. AB - OBJECTIVE: This study addresses test-retest reliability of the Postural and Repetitive Risk-Factors Index (PRRI) for work-related upper body injuries. This assessment was developed by the present authors. DESIGN: A repeated measures design was used to assess the test-retest reliability of a videotaped work-site assessment of subjects' movements. SUBJECTS: Ten heavy users of video display terminals (VDTs) from a local banking industry participated in the study. SETTING: The 10 subjects' movements were videotaped for 2 hours on each of 2 separate days, while working on-site at their VDTs. MAIN OUTCOME MEASURE: The videotaped assessment, which utilized known postural risk factors for developing musculoskeletal disorder, pain, and discomfort in heavy VDT users (ie, repetitiveness, awkward and static postures, and contraction time), was called the PRRI. The videotaped movement assessments were subsequently analyzed in 15 minute sessions (five sessions per 2-hour videotape, which produced a total of 10 sessions over the 2 testing days), and each session was chosen randomly from the videotape. The subjects' movements were given a postural risk score according to the criteria in the PRRI. Each subject was therefore tested a total of 10 times (ie, 10 sessions), over two days. The maximum PRRI score for both sides of the body was 216 points. RESULTS: Reliability coefficients (RCs) for the PRRI scores were calculated, and the reliability of any one session met the minimum criterion for excellent reliability, which was .75. A two-way analysis of variance (ANOVA) confirmed that there was no statistically significant difference between sessions (p < .05). Calculations using the standard error of measurement (SEM) indicated that an individual tested once, on one day and with a PRRI score of 25, required a change of at least 8 points in order to be confident that a true change in score had occurred. The significant results from the reliability tests indicated that the PRRI was a reliable measurement tool that could be used by occupational health practitioners on the job site. PMID- 9344309 TI - Rehabilitation of Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is the most common cause of acute neuromuscular paralysis in developed countries. GBS is a significant cause of new long-term disability for at least 1,000 persons per year in the United States, and more elsewhere. Given the young age at which GBS sometimes occurs and the relatively long life expectancies following GBS, it is likely that at least 25,000 and perhaps 50,000 persons in the US are experiencing some residual effects of GBS. Approximately 40% of patients who are hospitalized with GBS will require admission to inpatient rehabilitation. For GBS persons necessitating admission to inpatient rehabilitation, the requirement of prior ventilator support most strongly predicts an extended length of stay on inpatient rehabilitation. Other issues that affect rehabilitation are dysautonomia, cranial nerve involvement, and various medical complications associated with GBS. Deafferent pain syndrome is common in the early stages of recovery. Multiple medical complications, including deep venous thrombosis, joint contractures, hypercalcemia of immobilization, and decubitii, may develop in the early stages of recovery and interfere with the rehabilitation program. Anemia is a frequent finding in the first few months of illness but does not appear to interfere with functional recovery. Therapy should not overfatigue the motor unit, which has been associated with paradoxical weakening. Little is known of the long-term implications of the disability caused by GBS. Work similar to that performed for postpolio syndrome and spinal cord injury should be started in the rehabilitation setting. PMID- 9344310 TI - Herpes zoster polyradiculopathy. AB - Herpes zoster infection, resulting from reactivation of the dormant varicella zoster virus in the dorsal root ganglia, usually causes a painful dermatomal vesicular rash. Rarely, associated peripheral motor weakness is present, the mechanism of which is unclear. Three patients are reported who had focal limb muscle weakness associated with zoster infection. Physical and occupational therapy played a key role in motor function recovery of the patients, yet emphasis on the rehabilitation of postherpetic motor weakness is lacking in the literature. Physiatrists evaluating patients with limb muscle weakness following herpes zoster infection should be alert to this condition. The clinical syndrome of herpes zoster radiculopathy and the rehabilitation of these patients are discussed. PMID- 9344308 TI - Muscle metabolism changes with training in the nonamputated limb after vascular amputation: interest of phosphorus 31 NMR spectroscopy. AB - OBJECTIVE: To determine by 31P nuclear magnetic resonance (NMR) spectroscopy the efficacy of training in improving aerobic metabolism of calf muscle in nonamputated limb after recent vascular amputation; to assess the possible associated microcirculatory changes; and to evaluate the need for noninvasive monitoring techniques during training in the nonamputated limb after recent vascular amputation. DESIGN: Prospective study, before and after training. Subjects served as their own controls and were compared with a control group. SETTING: Rehabilitation center of a university hospital. PATIENTS: Ten unilateral vascular amputated patients were included with ankle systolic index between 0.5 and 0.8 in the nonamputated limb, and 10 control subjects without cardiovascular disease or risk factors of atherosclerosis with ankle systolic index of >.95. INTERVENTION: Walking with prosthesis at self-selected velocity over increasing walking distance, arm training at a workload of 60% of a maximal arm test, and analytical exercises of the nonamputated leg (dynamic contractions against low resistance). Subjects received training as inpatients, 5 days a week. MAIN OUTCOME MEASURES: Before and after training, ankle systolic index, forefoot transcutaneous oxygen tension (TcPO2) and veno-arteriolar reflex, and digital plethysmography of the second toe with reactive hyperemia test were studied. Changes in calf muscle pH, phosphocreatine (PCr), and inorganic phosphate (Pi) were measured by 31P NMR spectroscopy at rest and during a plantar flexion-type incremental protocol. RESULTS: There was no significant difference in ankle systolic index (.63 +/- .10 vs .64 + .07) or in TcPO2 (42 +/- 11 vs 44 +/- 10mmHg), and there was reappearance of veno-arteriolar reflex in 3 cases, of a plethysmographic signal in 2 cases, and of the positivity of the reactive hyperemia test in 3 cases. No differences were found with 31P NMR spectroscopy at rest before and after training. At the same workload (1 watt) the difference of the ratio (PCr/(PCr + Pi)) of rest to effort (PCr depletion) was significantly increased in the amputated patients (.423 +/- .159 vs .145 +/- .058; p < .01). This difference of ratio was lower after training (.360 +/- .158 vs .423 +/- .159; p < .05). The pH was less acid between the two periods. CONCLUSION: Vascular monitoring with systolic index and TcPO2 is necessary to follow and to prevent serious ischemia of the nonamputated limb. Claudication is often not detected because of early exhaustion during walking. Training after recent vascular amputation improves the skeletal muscle oxidative capacity. PMID- 9344311 TI - Delayed traumatic cerebral aneurysm after brain injury. AB - Traumatic aneurysms (TAs) are an unusual etiology for late neurological deterioration after traumatic brain injury (TBI) and represent less than 1% of all cerebral aneurysms. TAs most often are diagnosed acutely but may be delayed in presentation. To increase awareness of this serious but treatable condition when diagnosed early, we report a delayed TA after a motor vehicle accident. The patient experienced a seizure on day 46 postinjury while in rehabilitation and demonstrated persistent lethargy and hemiparesis. Neuroimaging revealed a large, ruptured left pericallosal artery TA, which was surgically clipped. The patient completed his rehabilitation course and was eventually discharged home with family. Among TBIs, TAs are associated with penetrating injuries and skull base or anterior cranial fossa fractures. Associated mortality is high, especially if rupture has occurred. Although TAs are rare, the clinician should be vigilant in the at-risk patient. PMID- 9344312 TI - Exploring the basis for Tai Chi Chuan as a therapeutic exercise approach. AB - For many centuries Tai Chi has been a martial art form, practiced primarily in Oriental cultures. For the past 300 years this movement approach has been used as an exercise form, practiced by millions of Chinese elderly people. To date, virtually no information exists about the therapeutic elements of this intriguing movement sequence. This article provides a historical review of existing documentation of reputed Tai Chi benefits. The 108 "forms" of Tai Chi Chuan are reduced to 10 composite forms for ease of application of these forms to older individuals within a reasonable time frame. An effort is set forth to identify the potential therapeutic elements within these forms. PMID- 9344313 TI - Statistical methods in rehab literature. PMID- 9344314 TI - Flat back and postpolio syndromes. PMID- 9344315 TI - Are we just specialists or is it time for us to broaden our horizons in treating the cancer patient? PMID- 9344316 TI - The epidemiology of soft tissue sarcoma. AB - Soft tissue sarcoma (STS) accounts for approximately 1% of all cancers diagnosed annually in the United States. Population-based data from Connecticut covering the years 1935-1989 have shown an increasing incidence of STS in both genders, with a greater increase among men than women. The recent increase in acquired immune deficiency syndrome-related Kaposi's sarcoma does not explain the upward trend in STS, dating back decades. Etiologic heterogeneity is suggested by epidemiologic variations that have been observed by subsite and cell type. Among the environmental factors associated with STS are external radiation therapy, Thorotrast, arsenical pesticides and medications, phenoxyherbicides, dioxin, vinyl chloride, immunosuppressive drugs, alkylating agents, androgen-anabolic steroids, human immunodeficiency virus, and human herpes virus type 8. In addition, STS occurs excessively among persons with certain heritable states including retinoblastoma, Li-Fraumeni syndrome, Gardner's syndrome, Werner's syndrome, nevoid basal cell carcinoma syndrome, neurofibromatosis type 1, and some immunodeficiency syndromes. These risk factors account for a minority of STS cases but provide leads for further epidemiologic and interdisciplinary studies into the genetic and environmental determinants of various forms of STS. PMID- 9344317 TI - Immunohistochemical and molecular genetic approaches to soft tissue tumor diagnosis: a primer. AB - During the past two decades we have witnessed the identification of an expanding list of immunohistochemical and molecular markers linked to histopathologically defined subtypes of tumors. These markers offer new insights and approaches to the classification of tumors with important prognostic and/or therapeutic implications. We review the potentially diagnostic immunohistochemical and molecular markers of soft tissue tumors (STTs). The immunohistochemical markers reviewed include vimentin, cytokeratin, desmin, HHF35, S100, myoD1, alpha1 antitrypsin, vascular markers (factor VIII, CD31, CD34), MIC2, and others. The potentially diagnostic chromosomal translocations and associated genes identified in STT include Ewing's/PNET t(11;22)(q24;q12)(FLI1;EWS), t(21;22)(q22;q12)(ERG; EWS); t(7;22)(p22;q12)(ETV1;EWS); desmoplastic small round cell tumor t(11;22)(p13;q12)(WT1;EWS); extraskeletal myxoid chondrosarcoma t(9;22)(q22;q12) (TEC(CHN);EWS); malignant ectomesenchymoma t(11;22)(q24;q12)(FLI1;EWS); alveolar rhabdomyosarcoma t(2;13)(q35;q14)(PAX-3;FKHR); t(1;13) (p36;q14)(PAX-7;FKHR); myxoid and round cell liposarcoma t(12;16)(q13;p11)(CHOP;TLS(FUS)); synovial sarcoma t(X;18)(p11;q11)(SSX1&2;SYT), and others. The nature, utility, and limitations of these markers in diagnostic settings are explored. PMID- 9344318 TI - Surgical management of soft tissue sarcomas. AB - Surgical therapy is an essential component of the treatment for soft tissue sarcomas. Although multimodality treatment has become more common for sarcomas, to achieve local control and potential cure, surgical resection of the primary tumor is necessary. The selection of the overall treatment regime for soft tissue sarcomas is predicated on tumor histology, grade, prior treatment, and to some extent on anatomic location. The surgical principles important for local tumor control are complete en bloc excision of the tumor, and pathology confirmation of tumor-free margins. The operative procedures vary with the anatomic location of the primary tumor, but the principle of pathologic free margins remains. Nonamputative resection is possible in the majority of extremity soft tissue sarcomas. Surgical resection of metastatic disease has been of value in selected patients. PMID- 9344319 TI - Treatment of cardiac tumors by orthotopic cardiac transplantation. AB - Primary soft tissue tumors of the heart usually cannot be excised with adequate margins. Orthotopic heart transplantation (OHT) could allow complete resection of cardiac tumors and has been performed in selected patients. However, most are not transplanted because of the high risk of tumor recurrence or metastasis and the possible enhancement of tumor growth by immunosuppressive drugs. Six patients with soft tissue cardiac tumors have been transplanted at the Columbia Presbyterian Medical Center: one paraganglionoma and one fibroma (both benign), and four malignant primary sarcomas. In all cases, there was no preoperative evidence of metastasis. In all but one case, the tumor was completely resected with adequate margins at the time of transplantation. One sarcoma patient who had not received preoperative or postoperative chemotherapy died suddenly 2.6 months after operation with no evidence of tumor. One patient is alive at 38 months with diffuse metastatic disease; two patients were treated preoperatively with intensive doxorubicin-based chemotherapy (one of whom also received postoperative external-beam radiotherapy to a positive surgical margin) and are tumor free 16 and 6 months after transplantation. Our experience compares favorably with the worldwide results of OHT for cardiac tumors (an additional 13 patients). PMID- 9344320 TI - Prognostic factors for local control of sarcomas of the soft tissues managed by radiation and surgery. AB - During the past decade, local control of primary sarcomas of the extremities by radiation and conservative surgery has supplanted more radical compartmental resections or amputations. Reviews of others and our published data show that the probability of achieving local control is highly dependent on achieving negative surgical margins. Other factors, such as pathological grade and size, histopathology, and concomitant chemotherapy may also affect local control, to a much lesser extent, although these are strongly correlated with the likelihood of distant metastatic disease. Appreciation of the importance of these different prognostic factors has been fundamental to the development of the current rationale for sarcoma management. PMID- 9344321 TI - Isolation limb perfusion with tumor necrosis factor alpha and chemotherapy for advanced extremity soft tissue sarcomas. AB - The unique property of high dose recombinant tumor necrosis factor alpha (rTNF alpha) is to activate and selectively destroy the tumor-associated microvasculature. For the systemic application of rTNF alpha it has been shown that the maximum tolerated dose (MTD) is 10 times less than the effective dose in animals. The main toxicity corresponds to systemic inflammatory response syndrome with a decrease in vascular resistance and hypotension. We found that it is possible to administer rTNF alpha at 10 times the MTD in an isolated limb perfusion (ILP) system with heart-lung machine, for locally advanced extremity soft tissue sarcomas. One hundred forty patients received an ILP with high-dose TNF alpha. In 55 patients treated with the combination of high-dose rTNF alpha + interferon-gamma + melphalan an overall objective response rate of 87% with 36% complete responses was observed; it was 81% and 28%, respectively, in a group treated with TNF alpha and melphalan (n = 85). Angiographic and immunohistological studies showed the selective and early damage of the sarcoma associated microvasculature preceded by the upregulation of adhesion molecules and intratumoral leak of von Willebrand factor. Tumor invasion by platelets and, in some cases, by polymorphonuclear cells, appeared within hours after the application of rTNFa long before the lysis of the tumor. Thus, ILP with high-dose TNF alpha and chemotherapy seems to act through a dual targeting: TNF hits the tumor associated vasculature, and chemotherapy attacks the tumor cells. Therefore, ILP with TNF is a new option in the management of locally advanced soft tissue sarcoma of the extremities. PMID- 9344322 TI - Adjuvant therapy of sarcomas of soft tissue. AB - Adjuvant chemotherapy for rhabdomyosarcomas, osteosarcomas, and Ewing's sarcomas is established, but remains controversial in other adult sarcomas. Of the 14 reported adjuvant studies, the largest study from the European Organization for Research and Treatment of Cancer showed a significant improvement in local control for adjuvant chemotherapy in nonextremity sarcomas, but not overall survival benefit. Two studies show a significant overall survival advantage for chemotherapy. A meta-analysis of individual patient data by the Medical Research Council Cancer Trials Office reported a trend but no significant difference in survival (although differences were significant for disease-free survival and local control). Adjuvant chemotherapy should not be offered for low-grade lesions because of their low probability of metastatic spread, nor should it be offered for small (< 5 cm) high-grade sarcomas because of their good prognosis. Oncologists should discuss the adjuvant data with patients who have soft tissue sarcomas. Patients informed of a difference in disease-free survival but a 4% absolute difference in survival may or may not wish to receive adjuvant chemotherapy. PMID- 9344324 TI - Chemotherapy for advanced sarcoma: therapeutic decisions and modalities. AB - In patients who have advanced soft tissue sarcoma that is no longer localized, systemic chemotherapy is the most reasonable option for treatment. The decision to treat or to use experimental or conventional agents should be based on the clinical assessment of anticipated net benefit in quality of life as well as the remote possibility of complete remission or even cure. Asymptomatic elderly patients with relatively stable disease might best be treated with watchful waiting; whereas younger excellent-performance-status patients should be offered the opportunity of participating in phase II or phase I studies of newer drugs and intensification regimens. Of the currently available single agents, only doxorubicin (or epirubicin) and ifosfamide show response rates greater than 20%; both show a definite dose-response relationship. Dacarbazine shows particular activity in uterine leiomyosarcomas. Combination chemotherapy regimens such as doxorubicin-ifosfamide show a higher response rate, but may be more toxic. New agents are needed. The current progress in our understanding of the molecular biology of sarcomas, and our expanded comprehension of the mechanism of action of cytotoxic drugs and the biology of drug resistance is cause for optimism. PMID- 9344323 TI - The role of chemotherapy in the management of non-metastatic operable extremity osteosarcoma. AB - Original articles published between 1991-1996 were selected according to specified criteria, and reviewed to provide answers to nine important questions about the role of chemotherapy in the management of non-metastatic extremity osteosarcoma: (1) Does adjuvant chemotherapy improve survival? (2) Are the results of the Rosen T10 protocol reproducible in different settings? (3) Is chemotherapy with two of the most active drugs (DOX/DDP) an effective adjuvant treatment, and comparable to other multiagent regimens? (4) Does histological response to neoadjuvant chemotherapy correlate with reduced local recurrence and/or improved survival? (5) Does a change of chemotherapy for patients whose tumors show a poor histological response to chemotherapy improve survival? (6) Does chemotherapy given before surgery (neoadjuvant) improve survival? (7) Are certain drugs, or their method of administration (route, duration, total dose, dose intensity, pharmacokinetics) important in determining outcomes? (8) Can new agents such as Ifosfamide be incorporated into intensive multi-agent chemotherapy, and does this improve pathological response and/or survival? (9) Can dose intensity of treatment be increased with G-CSF? The brief answers to questions 1-3 and 7-9 are "Yes"; question 4 "Yes, but may be changing"; and questions 5, 6 "Not proven," and these are expanded in the text. Future directions for treatment of osteosarcoma are covered under the headings identification of new agents, dose intensification, circumvention of drug resistance, immunotherapy, and insulin-like growth factor. PMID- 9344325 TI - Drug resistance in the treatment of sarcomas. AB - The antineoplastic action and development of drug resistance are reviewed for chemotherapeutic agents used in the treatment of sarcoma, including alkylating agents (cyclophosphamide, ifosphamide, dacarbazine), platinum compounds (cisplatin, carboplatin), the anthracycline compound doxorubicin, the topoisomerase II inhibitor etoposide, and the taxanes (paclitaxel, taxotere). Drug resistance mechanisms discussed include changes in intracellular glutathione and metallothione levels, increased aldehyde dehydrogenase levels, enhanced DNA repair and protection from apoptosis (for alkylating agents); increased 0-6 alkyltranferase levels (for dacarbazine); multidrug resistance 1- and multidrug resistance associated protein-mediated drug export from cells (anthracyclines, taxanes); and structural alteration of microtubules (taxanes). PMID- 9344326 TI - The future of sarcoma treatment. AB - The last two decades have shown exciting and dramatic improvements in the management of osteogenic sarcoma, while progress in soft tissue sarcomas has lagged behind considerably. Osteogenic sarcoma treatment has been a model of multidisciplinary collaboration. Orthopedic surgeons working together with medical and pediatric oncologists have improved disease-free survival while improving limb salvage rates and limb function. In contrast, the care of soft tissue sarcoma remains fragmented among many disciplines and specialties. Medical and pediatric oncologists, radiation oncologists, and a myriad of surgical specialists are all involved in the care of soft tissue sarcomas, and significant treatment (usually surgical) often occurs before referral to a center. Significant variation in managment leads to considerable difficulty in assessing the effects of treatment on outcome. Improvement in soft tissue sarcoma management will occur only when physicians recognize the need to centralize care to appropriate physicians within referral centers where patients can be treated on standardized cooperative protocols. PMID- 9344329 TI - Increased expression of functional Fas-ligand in activated T cells from patients with systemic lupus erythematosus. AB - The Fas ligand induces apoptosis upon binding to Fas/APO-1 (CD95) bearing target cells. Activation induced cell death (AICD) in T cells is mediated by upregulation of Fas ligand on the cell surface membrane upon crosslinking of the TCR. AICD is considered to be essential for the elimination of autoreactive T cells in the peripheral blood. To elucidate possible abnormalities in the process of AICD in human SLE, we studied the expression and function of Fas ligand in polyclonal T cell lines from patients with SLE, patients with other rheumatic diseases and normal controls. SLE T cells expressed on their surface significantly higher amounts of Fas ligand compared to the two control groups. Stimulation of the cells with anti-CD3 mAb lead to further increase in surface membrane Fas ligand expression in all three groups with SLE expressing the highest amounts. The percentage of increase was though lower in SLE T cells than in normal T cells or disease control cells. The T cells were examined for Fas ligand-mediated cytotoxicity in a 51Cr release assay using Fas-expressing normal T cells as target cells. There was no difference in SLE and control T cells with regard to specific 51Cr lysis, indicating that the Fas ligand expressed by the SLE T cells is functional. Our data show that activated T cells from patients with SLE express high amounts of functional Fas ligand with intact TCR-mediated upregulation. This could account for the high apoptotic rates that have been observed in lymphocytes from patients with SLE. PMID- 9344327 TI - Profiles of pulp infiltrating lymphocytes at various times throughout feather regeneration in Smyth line chickens with vitiligo. AB - Smyth line (SL) chickens develop a spontaneous, autoimmune, posthatch loss of pigment cells (vitiligo) in regenerating feather tissue. Smyth line vitiligo (SLV) is associated with lymphocyte infiltrations prior to and throughout the development of the disorder. It was the purpose of this study to determine the type, relative amounts, and proportions of pulp-infiltrating lymphocytes at various times throughout the growth of regenerating feathers. Feathers were plucked from 8-week-old chickens with and without SLV. Feather pulp cell suspensions were prepared when the regenerating feathers were 2, 3, 4, and 6 weeks of age. Cells were fluorescently labeled using a panel of mouse monoclonal antibodies specific for chicken lymphocytes. Both T and B cells infiltrated the feather pulp of chickens with SLV. T cell levels remained elevated throughout the 6 weeks of feather growth, while B cell levels steadily declined to control levels over the same time. The pulp-infiltrating cells were primarily T cells with an alphabeta T cell receptor expressing the Vbeta1 gene (TCR2+). The ratio between CD4+ and CD8+ cells was 1.42 and 0.75 in 2- and 6-week-old regenerating feathers from chickens with autoimmune SLV, respectively. In non-vitiliginous chickens this ratio was always near 1. These data suggest that TCR2+ T cells play an important role in SLV. CD4+ cells may play a recruiting/activating role, whereas CD8+ cells may have cytotoxic activity specifically directed against melanocytes. Additionally, this is the first report demonstrating the infiltration of B cells into the feather pulp of vitiliginous chickens. These B cells may directly/indirectly contribute to melanocyte destruction in SLV. PMID- 9344328 TI - Antibodies to the tyrosine phosphatase-like protein IA-2 are highly associated with IDDM, but not with autoimmune endocrine diseases or stiff man syndrome. AB - Antibodies to the 40 kD antigen (identified as tyrosine phosphatase IA-2) and glutamate decarboxylase (GAD65) are strongly associated with insulin dependent diabetes mellitus (IDDM). However, antibodies to GAD (GADA) can appear in the absence of IDDM, particularly in stiff man syndrome (SMS) and in some individuals with autoimmune polyendocrine syndrome type II (APS II) and organ specific autoimmune diseases. The aim of this study was to compare the specificity of IA-2 antibodies (IA-2A) and GADA for IDDM by determining their frequency in different patient groups. IA-2A were present in 64/114 (56%) IDDM patients and 9/19 (47%) APS II patients with IDDM but in only 4/28 (14%) SMS patients. 1/24 (4%) APS II patients without IDDM and 1/113 (0.9%) patients with organ specific autoimmune disease had low level IA-2A. In contrast GADA were present in 77/114 (68%) IDDM patients and 17/19 (89%) APS II patients with IDDM, but also in 25/28 (89%) SMS patients, 5/24 (21%) APS II patients without IDDM and 22/113 (19%) patients with organ specific autoimmune diseases. Furthermore, within the group of new onset IDDM, IA-2A seemed to be associated with ICA and age: 63% of ICA positive IDDM patients had IA-2A (74% had GADA) increasing to 77% in the group below 20 years of age (69% for GADA). Our results demonstrate that IA-2A may be more specific for IDDM than GADA, as the latter are also present in patients with SMS, APS II without IDDM and organ specific autoimmune diseases. IA-2A were less frequent in older patients with IDDM than GADA or ICA. A combination of IA-2A and GADA detected 84% of total and 93% of ICA positive IDDM patients. PMID- 9344330 TI - The effect of linomide, an immunoregulator in experimental autoimmune diseases, on humoral antibody responses in mice. AB - Linomide (quinoline-3-carboxamide), a well tolerated, orally administered compound was recently shown to be effective in the prevention and treatment of several autoimmune diseases in experimental animal models. We have investigated its effect on specific humoral immune responses directed to T-cell-dependent soluble or particulate antigens and to a T cell-independent antigen in several mouse strains. Linomide administered after antigen priming did not affect primary and secondary antibody responses directed to T-cell particulate antigens (SRBC) or soluble antigens given with or without complete Freund's Adjuvant (CFA). Linomide treatment given prior to antigen priming did not affect the antibody response to a soluble antigen (TNP-KLH) given with an adjuvant. In contrast, dose dependent down regulation of primary antibody responses was observed when T cell dependent (BSA-dextran) or T-cell-independent (TNP-Ficoll) antigens were administered in an immunogenic form without adjuvant after starting Linomide treatment. The primary anti-SRBC antibody response was also suppressed by high dose Linomide given prior to immunization although normal secondary responses were retained. It is worth noting that no immunosuppressive effects on antibody responses were found at low dose ranges which effectively reversed T cell dependent autoimmune manifestation. PMID- 9344331 TI - Humoral and cellular immune response to elastin in patients with systemic sclerosis. AB - The humoral immune response against elastin and collagen was studied in parallel with the delayed type hypersensitivity (DTH) reaction to elastin and the percentage of lymphocyte subpopulations in peripheral blood in 20 patients with systemic sclerosis (SSc). An increase of anti-elastin antibodies of all subclasses was found with a significant prevalence of IgE and IgA antibodies. The profile of anti-collagen type I and type IV antibodies showed an increase of IgE isotypes. In 25% of the patients (5 out of 20) positive DTH reactions to elastin were observed as compared to the negative skin reactions in all control individuals. At the same time a significant hyporeactivity to common bacterial and mould antigens was found in 40% of the patients (versus 16% in the control group) which could be an explanation for the low incidence of positive anti elastin DTH reaction. The DTH hyporeactivity in SSc cases was in contrast with the increased percentage of CD4 T cells (58.4 vs. 42.0) and increased CD4/CD8 ratio (2.5 vs. 1.5) in the peripheral blood of the patients. This finding together with the increased IgE antibodies to elastin and collagen type I and type IV might suggest a possible shift of the immune balance towards the Th2 type of immune response. This is in line with the increased CD8+CD57+ cells which correlated with the highest number of other parameters studied - disease duration, total skin score, IgE anti-elastin antibodies, IgG anti-collagen type I antibodies, CD4/CD8 ratio and CD19 B cells. The results of this study demonstrated the existence of both humoral and cell-mediated immune response against elastin in SSc patients. However, we could not define whether this was an essential part of pathogenetic mechanisms or a secondary phenomenon reflecting the extent of the damage of connective tissue. PMID- 9344332 TI - Antinuclear antibodies in children with chronic nonspecific complaints. AB - Children who are chronically complaining nonspecific symptoms such as headache, fatigue, abdominal pain, and low grade fever are commonly seen in daily pediatric outpatient clinics. Some of them are unable to go to school and are diagnosed as school refusal by physicians or educational staff. On the other hand, there are children who do not fulfill any criteria of collagen diseases and whose anti nuclear antibodies (ANA) are found to be positive. Some of these children have chronic nonspecific complaints. We prospectively studied the prevalence of ANA in children who visited a pediatric outpatient clinic because of chronic nonspecific complaints. Surprisingly, 74 out of 140 symptomatic children (52.4%) were positive for ANA, while only 5 out of 82 healthy control children (6.1%) were positive (p < 0.0001). 39 of 74 ANA positive patients (52.1%) have low ANA titers < or = 1:80, nevertheless 36 patients (47.9%) have high ANA titers > or = 1:160. ANA fluorescent patterns were homogeneous and speckled in 75.3%, speckled in 17.6% and others in 6.8%. ANA positive patients tended to have general fatigue and low grade fever, while gastrointestinal problems such as abdominal pain and diarrhea and orthostatic dysregulation symptoms were commonly seen in ANA negative patients. Children who were unable to go to school more than 1 day a week were seen significantly more in ANA positive patients than in negative patients. Autoantibody analysis using Western immunoblot revealed that 26 out of 63 ANA positive sera (41.3%) had antibodies to the 62 kD protein which had not been previously noticed. These data suggest that autoimmune mechanism may play a role in childhood chronic nonspecific symptoms. We therefore propose a new disease entity of the autoimmune fatigue syndrome in children. When chronically complaining children visit a pediatric out-patient clinic, immunological approaches should be considered before they are discriminated as school refusal or having psychogenic disorders. PMID- 9344333 TI - Evidence for the viral aetiology of IDDM. PMID- 9344334 TI - DNA immunization with a bovine rotavirus VP4 gene induces a Th1-like immune response in mice. AB - Immunization with naked plasmid DNA effectively induces both humoral and cell mediated immunity to vaccine antigens and can confer protection against numerous infectious diseases. To explore the potential use of DNA immunization to induce rotavirus-specific immune responses, we used plasmid DNA encoding the VP4 gene of bovine rotavirus (BRV). Intrasmuscular injection of the plasmid encoding the VP4 gene into C57BI/6 mice induced cell-mediated immunity as measured by cytokine production. Although DNA immunization did not induce a detectable BRV-specific antibody response, DNA-immunized animals were primed for antibody production and a cellular immune response. Following viral inoculation, the immunized animals displayed an enhanced number of BRV-specific antibody-secreting cells and cytotoxic activity. The immune response induced by DNA immunization alone or followed by viral inoculation was biased toward IFN-gamma production (Th1-like). CD4+ lymphocytes were the major source of IFN-gamma production in the spleen following DNA immunization. In contrast, a balanced cytokine production was observed in the spleens of animals receiving whole virus. These experiments showed that DNA immunization with a gene encoding the VP4 protein of BRV stimulated a Th1-like immune response in mice, and this bias in the immune response persisted following exposures to whole virus. PMID- 9344335 TI - Protective immunity elicited by vaccination with DNA encoding for a B cell and a T cell epitope of the A/PR/8/34 influenza virus. AB - Numerous reports have demonstrated that immunization with plasmids bearing influenza virus hemagglutinin (HA) or nucleoprotein (NP) genes elicits humoral and cellular protective responses. Herein we describe the generation of a plasmid (pVH-TB) encoding for a VH region of a self-Ig in which both the major B cell epitope HA150-159 and the immunodominant CD4 T cell epitope HA110-120 of HA of the A/PR/8/34 influenza virus were genetically inserted in the CDR2 and CDR3 loops, respectively. Our results demonstrate unequivocally that i.m. injection of pVH-TB plasmid in BALB/c mice elicited specific cellular and humoral immune responses able to protect against infection with lethal doses of A/PR/8/34 influenza virus. PMID- 9344336 TI - Detection of antibodies to a recombinant gag protein derived from human endogenous retrovirus clone 4-1 in autoimmune diseases. AB - To investigate whether human endogenous retroviruses (HERV) contribute to autoimmune diseases, we prepared a recombinant p30gag protein derived from clone 4-1 of the HERV family, using a baculovirus-vector system. This p30gag protein (CA41B) was approximately 30 kDa, as expected, and reacted with antibodies for p30gag purified from both murine and feline leukemia virus. This result suggested that the antigenic determinant for p30gag was well conserved in CA41B. Analysis of serum antibodies to p30gag in patients with autoimmune diseases was done by Western blotting. CA41B detected anti-p30gag antibodies in 48.3% of systemic lupus erythematosus (SLE) patients, 35.0% of Sjogren's syndrome (SS) patients, and 33.3% of mixed connective tissue disease (MCTD) patients, whereas no anti p30gag antibodies were found in healthy subjects. This suggested that HERV p30gag or other retroviral p30gag proteins possessing the same antigenic determinant as CA41B may play a role in these diseases. Although detection of antibodies to HERV p30gag in autoimmune diseases is indirect evidence that HERV proteins are involved, this study showed that patients with autoimmune diseases have antibodies to HERV p30gag using a recombinant HERV protein rather than synthetic peptides based on HERV or retroviral proteins of other species. PMID- 9344337 TI - Kinetics study of human retrovirus antigens expression in T lymphocytic cell lines by indirect immunofluorescence assay. AB - Indirect immunofluorescence assay (IFA) is a well-accepted assay for the confirmation of human retrovirus infection. Fluctuations in HIV-1 antigen expression in infected E-B2 cells depending on several factors have been reported. Cells kept in log phase expressed the highest levels of viral antigen. Thus, we studied the time kinetics of IFA positivity in MT-2 (HTLV-I), MO-T (HTLV II), CEM, and H9 (HIV-1) cell lines. Uninfected T cell line, HT, was used as nonspecific control. Reference HTLV-I/II and HIV-1 serum panels from the Centers for Disease Control and Prevention (CDC) were tested by conventional IFA procedure on slides of each cell line made on different days. On the second day after subculture, HTLV-I strongly positive sera reacted on MT-2 and MO-T cells with a bright pericytoplasmic fluorescence pattern. Weakly positive sera showed a faint staining from the fifth day on, when all the sera showed the highest degree of fluorescence. With HIV-1 cell lines, sera predominantly reacted with a diffuse cytoplasmic pattern, although some sera showed a granular and pericytoplasmic capping staining. The highest degree of fluorescence was found at 3-5 days after subculture. We demonstrated that the sensitivity of the IFA for the detection of antibodies against human retroviruses depended on the day when the slides were assayed and on the serum antibody titer. The fifth day was the most appropriate for HTLV-I/II and HIV-1/H9 systems, whereas for HIV-1/CEM, the fourth day was better. Furthermore, the intensity of the immunofluorescence pattern differed with the antibody titers and the level of antigens expressed on the four cell lines studied. The IFA, improved in our laboratory, proved to be very sensitive, specific, and rapid and could be used as a supplementary/confirmatory assay for retrovirus infection. PMID- 9344338 TI - Characterization of several pseudorabies viral strains by virus-neutralization test using monoclonal antibodies. AB - In the present study, five mouse monoclonal antibodies (MAbs) to the pseudorabies virus (PRV) Yamagata-81 strain were produced. The MAbs were used in cross neutralization tests and cross-indirect enzyme-linked immunosorbent assay (ELISA) against three PRV viral strains isolated in Argentina and another four obtained from the United States, Japan, France, and Sweden. Four of five MAbs needed the presence of complement to produce or enhance neutralization activity. No differences were observed by ELISA. The MAbs showed different neutralizing activity against PRV strains, suggesting phenotypic heterogeneity among them. PMID- 9344340 TI - Characterization of paracellular penetration routes. PMID- 9344339 TI - Immunoglobulin M antibody response to measles virus following natural virus infection, primary vaccination, and reexposure to the virus. AB - Evaluation of the measles virus ELISA kit (Merck) to detect specific IgM as an indicator of primary measles antibody response was carried out. A modification of the manufacturer's cutoff value interpretation was introduced to allow for equivocal results in addition to positive and negative ones. With this modification, the test assayed gave an overall reproducibility of 96.16%. The IgM seropositivity rate for seroneutralization-confirmed measles cases was 100% for naturally infected measles subjects and 90% for primary measles vaccinated subjects. Individuals with positive neutralizing antimeasles antibodies in close contact with a confirmed measles case gave the following measles IgM ELISA results: 54.54% negative, 9.09% positive, and 36.36% equivocal, showing a booster with IgM antibody response on reexposure to the virus. Positive subjects with neutralizing antimeasles antibodies without recent contact with a measles case gave negative IgM results. IgM seropositivity was strongly associated with IgG seroconversion and clinical measles (p < 0.0001). The technique assayed performed adequately for the confirmation of both measles natural infection and primary vaccination and for the differentiation of primary and secondary antibody response, taking into account the modification in the cutoff value interpretation introduced and providing that the serum samples are obtained between days 5 and 30 after onset of rash. PMID- 9344341 TI - Introduction of Davida Y. Teller 1997 recipient of the Friedenwald Medal. PMID- 9344342 TI - First glances: the vision of infants. the Friedenwald lecture. PMID- 9344343 TI - Pathology of macular lesions from subnanosecond pulses of visible laser energy. AB - PURPOSE: To demonstrate how current theories regarding ultrashort laser pulse effects may apply to ocular tissue, a prospective clinicopathologic study of macular lesions from ultrashort laser pulses compared the pathologic effects with the clinical and fluorescein angiographic appearance of the laser lesions. METHODS: Ninety-femtosecond, 3-picosecond, and 60-picosecond laser pulses, throughout a range of energies, were delivered to the retina of Macaca mulatta. Clinical examination and fluorescein angiography were performed at 1 hour in all eyes and 24 hours after exposure in selected eyes. Eyes were enucleated at 1 or 24 hours after lesion placement. The structure and extent of retinal lesions were scored for comparison with the clinical findings. RESULTS: Focal retinal pathologic appearance correlated well with a clinically visible lesion observed 24 hours after laser delivery. Retinal lesions were small foci of retinal pigment epithelium (RPE) and retinal disruption, without choriocapillaris involvement. Lesions that contained focal RPE vacuoles or lifting of the RPE also demonstrated leakage, in fluorescein angiographic studies. Suprathreshold laser delivery frequently caused focal columns of retinal injury and intraretinal hemorrhages from retinal vessel bleeding, with no rupture of choroidal blood vessels. CONCLUSIONS: The retinal response to ultrashort laser pulses at moderate energy followed a pattern of focal damage from laser-induced breakdown without significant thermal spread. PMID- 9344344 TI - Prostaglandins alter extracellular matrix adjacent to human ciliary muscle cells in vitro. AB - PURPOSE: This study investigates the possibility that prostaglandins (PGs) induce changes in extracellular matrix (ECM) adjacent to ciliary muscle cells. METHODS: Human ciliary smooth muscle cells were grown to confluence in monolayer cultures and were treated with PGF2alpha, 11-deoxy-PGE1, or PhXA85 (the nonesterified analogue of PhXA41) for 12 to 72 hours. The amount of collagens type I, III, and IV in the cultures was determined, using sandwich enzyme immunoassays. The distributions of these collagens were assessed in the PG-treated cultures by immunocytochemistry. RESULTS: Twenty-four-hour treatment with 20 nM PGF2alpha, 11 deoxy-PGE1, or PhXA85 reduced the amount of collagen type I in extracts of the cell layer by 65+/-10%, 56+/-7%, and 46+/-7%, respectively, when compared with levels of those substances in vehicle-treated cultures. In similar fashion, collagen type III in cell layer extracts was reduced by 41+/-5%, 33+/-9%, and 3+/ 5%, respectively. When the concentration of PGs was increased to 200 nM, the amount of type III collagen in the cell layer extracts was reduced by 93+/-7%, 99+/-1%, and 99+/-1%, respectively. Changes in type IV collagen in cell layer extracts after treatment with 20 nM PGs were not statistically significant. When the concentration of PGF2alpha, 11-deoxy-PGE1, or PhXA85 was increased to 200 nM, the amount of collagen type IV in the cell layer extract increased by 101+/-16%, 14+/-5%, and 89+/-11%, respectively. There were minimal changes in the staining pattern for collagen type I after 24-hour treatment with 20 nM PGs. When the PG concentration was increased to 200 nM, there were reductions in the density of collagen type I fibrils and clumping of collagen type III immunoreactive elements. The delicate lacework of collagen type IV immunoreactivity was replaced by bundles or clumps of heavy immunoreactive strands, separated by areas without immunoreactivity. These changes were present in cultures exposed to 20 nM PGs and were marked when PG concentration was increased to 200 nM. CONCLUSION: These results indicate that PGs can induce substantial changes in the ECM around ciliary smooth muscle cells in vitro. These data support the possibility that changes in ciliary muscle ECM may contribute to increased uveoscleral outflow facility after topical PG administration. PMID- 9344345 TI - Cell-mediated immune response and stability of intraocular transgene expression after adenovirus-mediated delivery. AB - PURPOSE: To evaluate the role of cell-mediated immunity in the stability of ocular adenovirus-mediated transgene expression. METHODS: Adenovirus (4 x 10(6) pfu) containing lacZ (Ad.CMVlacZ) was injected intravitreally or subretinally into one or both eyes of immunocompetent (+/+) and immunocompromised (nu/nu) CD-1 mice. Control eyes received injections of saline. Additional +/+ mice received subretinal injections of Ad.CMVlacZ with coadministration of 200 microg of human immunoglobulin (Ig) G or CTLA4Ig by intraperitoneal, intravitreal, or subretinal injection. The mice were killed at various times after injection, and their eyes were examined histologically and immunohistochemically. RESULTS: LacZ expression was extended from 1 week to more than 5 weeks in the corneal endothelium, iris, and trabecular meshwork of nu/nu mice compared with time of expression in +/+ mice when adenovirus was administered intravitreally. In contrast, subretinal injection resulted in only a minimal increase in transgene stability in nu/nu mice compared with that in +/+ mice. Neither systemic nor intraocular administration of IgG or CTLA4Ig affected the stability of lacZ expression in the retina or retinal pigment epithelium after subretinal injection in +/+ mice. Immunoglobulin G and CTLA4Ig enhanced the stability of transgene expression in the trabecular meshwork. CONCLUSIONS: A T-cell-mediated immune response appears to play a role in the loss of adenovirus-mediated lacZ expression after intravitreal but not after subretinal injection. This result implies that the subretinal space is an immune-privileged site and a favorable site for gene transfer. PMID- 9344346 TI - Cultivation of rabbit corneal epithelial cells in serum-free medium. AB - PURPOSE: To establish conditions for cultivation, serial growth, and normal differentiation of corneal epithelial cells in serum-free medium (SFM). METHODS: Rabbit corneal epithelial cells were co-cultured with lethally treated 3T3-cell feeder layers. Instead of serum, medium was supplemented with serum albumin, hormones, and other additives. Cell growth was quantitated spectrophotometrically with a new rhodamine-B staining protocol with a sensitivity range of 5 X 10(3) to 1 x 10(5) cells/cm2. Keratin expression was analyzed by immunostaining or sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cell extracts. RESULTS: In SFM, without growth factors, cells grew no more than six to eight doublings, but when 10 ng/ml epidermal growth factor were added, serial transfer was possible, and epithelial cells grew to up to 18 to 20 doublings (three cell passages). Two cell colony types were seen: One type was composed of nonstratified proliferating cells, and the other of stratified cells expressing high levels of the differentiation-linked keratins K3 and K12. Confluent cultures formed a four- to five-layer stratified epithelium whose suprabasal cells were stained with anti K12 antiserum. Acidic and basic fibroblast growth factors and epidermal growth factor reduced the expression of keratins K3 and K12. Transforming growth factor alpha and epidermal growth factor led to the highest stimulation of cell proliferation. Limbal, peripheral, and central corneal epithelial cells showed similar clonal growth abilities, but colony size was larger for cells derived from limbal epithelium. CONCLUSIONS: These SFM conditions support the serial transfer, normal differentiation, and formation of typical corneal epithelium by cultured corneal epithelial cells and are useful in studying and assaying a variety of cytokines and compounds that modulate corneal epithelial cell proliferation and differentiation. PMID- 9344347 TI - Blood-borne signals that induce anterior chamber-associated immune deviation after intracameral injection of antigen. AB - PURPOSE: Anterior chamber-associated immune deviation (ACAID) is elicited by an antigen-specific signal that escapes the antigen-containing eye and travels through the blood to the spleen. Two types of ACAID-inducing signals have been described: those associated with blood-borne monocytes, and a soluble factor found in serum. The authors sought to understand the basis for the existence of two distinct types of ACAID-inducing signals. METHODS: Different kinds of antigens (soluble, cell associated, particulate) were injected into the anterior chamber (AC) of normal, presensitized, and immunodeficient mice. In addition, peritoneal exudate cells were pulsed in vitro with different kinds of antigen in the presence of transforming growth factor beta and then evaluated for the ability to induce ACAID in naive (nonsensitized) as well as T- and B-cell deficient recipients. RESULTS: Among antigens injected into the AC, inert particulate antigens could not induce ACAID, but soluble and cell-associated (minor histocompatibility) antigens generated cell-associated ACAID-inducing signals. In contrast, antigens injected into the AC of presensitized mice generated ACAID-inducing signals that were soluble and located in the plasma fraction of blood. All ACAID-inducing signals created in vitro with soluble, particulate, or cell-associated antigens induced ACAID in vivo. CONCLUSIONS: Cell associated ACAID-inducing signals are generated in naive mice regardless of the kind of antigen, and these signals arise from mobile intraocular antigen presenting cells. However, when antigen is injected into the AC of presensitized mice, a soluble signal emerges, perhaps derived from T cells that enter the antigen-containing eye. Together, these signals dictate that subsequent exposures to ocular antigen will not evoke immunogenic inflammation. PMID- 9344348 TI - A new method of culturing and transferring iris pigment epithelium. AB - PURPOSE: To optimize a culture technique and transfer iris pigment epithelial (IPE) cells for cellular studies in vitro. METHODS: Porcine iris tissues were obtained, and IPE cells were isolated and cultured at high densities by plating them in the form of drops. Spherically shaped structures containing a high concentration of cells were formed after 7 to 10 days of culture. Cells were subcultured by transferring spheres to new culture dishes without employing enzymatic dissociation. The purity of IPE cells was determined by pigmentation and cytokeratin labeling. Proliferation was assessed by incorporation of 5-bromo 2'-deoxyuridine. Cellular structure was analyzed under the light and electron microscopes and function was assayed by rod outer segment phagocytosis. RESULTS: Iris pigment epithelial cells, when cultured at high densities, tended to form elevated spherical structures containing viable cells. The cultured cells were pigmented and showed positive labeling with a monoclonal cytokeratin antibody. The IPE cells proliferated and migrated from the spheres to form monolayers. Cells originating from the transferred spheres also continued to proliferate and to migrate in a similar manner to the originally cultivated cells to form monolayers after 7 to 10 days. These cells were able to phagocytose rod outer segments. CONCLUSIONS: This new method provides a simple method of culturing a large quantity of IPE cells. The high yield of pure IPE cells and the ease of transfer provide an ideal means to study them at the cellular level. PMID- 9344350 TI - Which method of flicker perimetry is most effective for detection of glaucomatous visual field loss? AB - PURPOSE: The authors compared the efficacy of two different forms of flicker perimetry: temporal modulation perimetry (TMP), which measures contrast thresholds for a fixed temporal frequency, and critical flicker frequency (CFF), which measures the highest frequency for which flicker is detected at a fixed contrast. METHODS: The authors compared 16 patients with early to moderate glaucomatous visual field loss with 16 age-matched normal controls. Flicker stimuli consisted of 2 degrees diameter targets of 2 seconds in duration, presented in 44 locations throughout the central 30 degrees visual field. Flicker was presented within a cosine envelope to avoid temporal transients. For TMP, contrast sensitivity thresholds were measured for 8-Hz sinusoidal flicker; CFF thresholds were measured for a stimulus of 100% contrast. RESULTS: The results indicate that TMP and CFF produced similar test-retest reliability in normals. CFF had slightly better reliability in glaucoma patients. Receiver operating characteristic analysis revealed that TMP could provide better separation of normals and glaucoma patients than did CFF. Similar findings were obtained when the thresholds for both procedures were converted to Z scores. CONCLUSIONS: Both methods of flicker perimetry testing provide acceptable test-retest reliability, and both can distinguish normal subjects from glaucoma patients. However, TMP is more effective in separating normal subjects from glaucoma patients than CFF, suggesting that TMP is the method of choice for detecting glaucomatous damage using flicker perimetry. PMID- 9344349 TI - Modulation by neurogenic acetylcholine of nitroxidergic nerve function in porcine ciliary arteries. AB - PURPOSE: To determine whether nitroxidergic, cholinergic, and vasoactive intestinal polypeptide (VIP)-mediated nerves participate in the regulation of porcine ciliary arterial tone and to analyze the mechanisms underlying the neuronal interaction. METHODS: Changes in isometric tension were recorded in helical strips of the arteries, which were stimulated by transmurally applied electrical pulses or nicotine. The presence of perivascular nerve fibers containing reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase, acetylcholinesterase, and VIP immunoreactivity were determined histologically. RESULTS: Transmural electrical stimulation (2, 5, and 20 Hz) and nicotine produced a relaxation of the arterial strips denuded of the endothelium and contracted with prostaglandin F2alpha. The response was not influenced by timolol but was abolished by oxyhemoglobin and methylene blue. N(G)-nitro-L arginine, a nitric oxide (NO) synthase inhibitor, abolished the neurogenic relaxation, and L-arginine restored the response. Physostigmine inhibited, but atropine potentiated, the neurogenic response. The relaxation was attenuated by acetylcholine but was not influenced by VIP. There were nerve fibers and bundles containing NADPH diaphorase, acetylcholinesterase, and VIP immunoreactivity in the adventitia of ciliary arteries. CONCLUSIONS: Porcine ciliary arteries are innervated by NO synthase-containing nerves that liberate NO, possibly as a neurotransmitter on excitation to produce muscular relaxation. Nitroxidergic nerve function is inhibited by acetylcholine released from cholinergic nerve, possibly because of impaired production or release of NO. VIP does not seem to function as a neurotransmitter or a modulator. PMID- 9344351 TI - Color vision defect type and spatial vision in the optic neuritis treatment trial. AB - PURPOSE: To describe the types of color vision defects present in the acute phase of the disease and 6 months into recovery in the 438 participants of the Optic Neuritis Treatment Trial. METHODS: Patients meeting strict eligibility criteria were seen within 8 days of the onset of symptoms and then at regular follow-up visits. At the first and 6-month visits (and subsequent annual visits), spatial vision (acuity, contrast sensitivity), visual fields, and color vision were measured. Farnsworth-Munsell 100-hue tests were scored by a variant of the method of quadrant analysis described by Smith et al (Am J Ophthalmol. 1985; 100:176 182). RESULTS: Most persons show mixed red-green (RG) and blue-yellow (BY) color defects (one type predominating, accompanied by a lesser defect of the other type). BY defects tend to be slightly more common in the acute phase of the disease, with slightly more RG defects at 6 months. Persons may shift defect type over time. Defect type was not related to any of the spatial vision measures at either test time or to treatment group; however, severity of color defect was related to both spatial vision measures and treatment group. CONCLUSIONS: Contrary to common clinical wisdom, optic neuritis is not characterized by selective RG defects. Color defect type cannot be used for differential diagnosis of optic neuritis. PMID- 9344352 TI - Current keratoconus detection methods compared with a neural network approach. AB - PURPOSE: Four videokeratographic methods for keratoconus detection were compared with a neural network approach. METHODS: A classification neural network for keratoconus screening was designed to detect the presence of keratoconus (KC) or keratoconus suspects (KCS); a separate cone severity network graded the severity of conelike topography patterns consistent with KC or KCS. Three hundred TMS-1 examinations (Tomey) were randomly divided into training and test sets. Ten topographic indexes were network inputs. Nine categories were used: normal, astigmatism, KC, KCS, contact lens-induced warpage, pellucid marginal degeneration, photorefractive keratectomy, radial keratotomy, and penetrating keratoplasty. KC was subdivided into KC1 (mild), KC2 (moderate), and KC3 (advanced). There were three outputs for the classification network (KC, KCS, and OTHER); target output values of 0 = OTHER, 0.25 = KCS, 0.5 = KC1, 0.75 = KC2, and 1.0 = KC3 were used for the severity network. RESULTS: The best-trained classification network had 100% accuracy, specificity, and sensitivity for the test set. The severity network had mean outputs (+/-standard deviation) of OTHER = 0.02+/-0.02, KCS = 0.21+/-0.05, KC1 = 0.52+/-0.17, KC2 = 0.74+/-0.12, and KC3 = 0.91+/-0.15. The severity network output for all categories was well correlated to the keratoconus prediction index (R = 0.892, P < 0.0001). The classification network had an overall accuracy and specificity significantly better (P < or = 0.005) than the Klyce/Maeda keratoconus index (KCI) test, the Rabinowitz test (K & I-S), and simulated keratometry (average Sim K). However, there were no significant differences in keratoconus sensitivity between the classification network, KCI, and K & I-S. The sensitivity and specificity of average Sim K were significantly worse than those of the other tests. The classification network had significantly better sensitivity (P < 0.001) and specificity (P = 0.025) for KCS detection than the K & I-S. CONCLUSIONS: The neural networks completely distinguished KC from KCS and from topographies that resembled KC. The network approach equaled the sensitivity of currently used tests for keratoconus detection and outperformed them in terms of accuracy and specificity. PMID- 9344353 TI - Calcium homeostasis of isolated single cortical fibers of rat lens. AB - PURPOSE: To investigate the calcium homeostasis in single fiber cells isolated from rat ocular lens cortex and to quantify the changes in the concentration of free intracellular calcium [Ca2+]i during the process of disintegrative globulization. METHODS: Individual fiber cells from the cortex of the adult rat lens were isolated by treatment with trypsin in ion-free buffered sucrose. The isolated fiber cells were loaded with the acetoxymethyl esters of Fluo-3 or Calcium Green-2, or with Fluo-3 and Fura Red, and changes in [Ca2+]i of single cortical fibers were measured using a microfluorometer. The time course of increase of [Ca2+]i in fiber cells exposed to Ringer's solution was measured, and the effects on the increase of [Ca2+]i of calcium channel blocker, verapamil, Na Ca exchange inhibitors Ni2+ and Zn2+, and protease inhibitor, leupeptin, Na+-free and K+-free media and Ca2+-containing isotonic sucrose solution, were investigated. RESULTS: In Hepes sucrose solution (containing approximately 1.5 microM Ca2+), the isolated fiber cells maintained stable values of [Ca2+]i at 99.6+/-10 nM (n = 32). Exposure of the isolated fibers to Ringer's solution (containing 2 mM Ca2+) led to a monoexponential increase of [Ca2+]i at a rate of 0.12 min(-1). This increase in [Ca2+]i was accompanied by disintegration of the isolated fibers into discrete but resealed globules. Changes in [Ca2+]i, monitored by using a two-dye ratiometric method using Fura Red and fluo-3, showed a progressive increase in [Ca2+]i in fibers exposed to Ringer's solution, preceding globulization. The [Ca2+]i in the globules in Ringer's solution, determined using Calcium Green-2, was 3.6+/-0.7 microM (n = 23). Compared with that in fibers in Ringer's solution, the rate of increase of [Ca2+]i in fibers was much slower in the presence of 50 microM verapamil (0.047 min[-1]), in Na+ free (0.086 min[-1]) and in K+-free (0.062 min[-1]) Ringer's solution, or when the fibers were suspended in Hepes-sucrose solution, containing 2 mM Ca2+ (0.046 min[-1]). After 30 minutes, the [Ca2+]i of fiber cells exposed to Ringer's solution, containing 2 mM Ni2+ (574.7+/-29 nM; n = 7) or Zn2+ (402.6+/-77 nM; n = 7) was significantly lower (P < 0.001) compared with that in fiber cells exposed to Ringer's solution alone (1995+/-461 nM, n = 10). In Ringer's solution, leupeptin delayed globulization without significantly affecting the increase in [Ca2+]i. The [Ca2+]i of fiber cells isolated from outer and inner cortex and suspended in Hepes-sucrose was comparable; however, after 15 minutes of exposure to Ringer's solution, [Ca2+]i in fibers from the outer cortex was approximately three times higher than [Ca2+]i in those from the inner cortex. CONCLUSIONS: Exposure to high (millimolar) concentrations of calcium in the external medium leads to an increase in [Ca2+]i of isolated individual fiber cells, which precedes disintegrative globulization. The protective effects of Na+-free and K+ free solutions on globulization appear to be due to a lower rate of increase of [Ca2+]i. Part of the calcium influx may be mediated by L-type calcium channels and by Na-Ca exchange, operating in reverse. Proteolytic inhibitors do not affect the increase in [Ca2+]i but delay globulization by inhibiting calcium-mediated proteolysis. The isolated fiber cells and the disintegrated globules maintain a 100- to 300-fold gradient of calcium across their plasma membranes. PMID- 9344354 TI - Role of small GTP-binding proteins in lovastatin-induced cataracts. AB - PURPOSE: To investigate the biochemical mechanisms involved in the cataract induced by lovastatin, a commonly used cholesterol-lowering agent. METHODS: The effects of lovastatin on lens transparency and on lens epithelial cell proliferation and structure have been investigated using organ-cultured rat lenses and cultured epithelial cells from human and rabbit lenses, respectively. Lens histologic and morphologic changes were recorded microscopically. Small GTP binding protein profiles were determined by [alpha-32P] GTP overlay assays. RESULTS: Rat lenses organ cultured for 7 days with lovastatin, a 3-hydroxy-3 methylglutaryl CoA reductase inhibitor, developed frank subcapsular opacity. Lens epithelial cells (both human and rabbit) demonstrated extensive morphologic changes and inhibition of proliferation when treated with lovastatin. Histologic sections of lovastatin-treated lenses showed partial to complete degeneration of the central epithelium, distortion of elongating epithelial cells, and extensive vacuole formation in the equatorial regions of the cortex. Supplementation of the medium with DL-mevalonic acid (a precursor of isoprenoids whose synthesis is inhibited by lovastatin) prevented the lovastatin-induced changes in whole lenses or in lens epithelial cell cultures, whereas supplementation with cholesterol had no such effect. GTP-binding proteins accumulated in the soluble fractions of lovastatin-treated lens epithelial cells. This was consistent with a blockade in isoprenylation preventing normal association with membranes. CONCLUSIONS: The findings suggest that impairment of the function of small GTP-binding proteins, due to a lovastatin-induced blockade in their isoprenylation, affects lens cell structure and proliferation in tissue culture and induces lens opacity in organ culture. These findings are consistent with the proposed roles of small GTP binding proteins as molecular switches that regulate fundamental cellular processes, including growth, differentiation, and maintenance of cell structure. PMID- 9344355 TI - Apoptotic cell death in rabbit lens after lens extraction. AB - PURPOSE: To determine whether apoptosis or necrosis of lens epithelial cells occurs after lens extraction. METHODS: Lens extraction was performed on 24 rabbit eyes. The authors then performed terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and transmission electron microscopy on the eyes 1 to 10 days after surgery. RESULTS: The postoperative lens was demarcated into three regions by the adhesion between the incised edge of the anterior capsule and the posterior capsule: the adhesive region, the intracapsular region, and the central portion of the posterior capsule. By day 2, TUNEL-positive cells with morphologic characteristics of necrosis were detected in the intracapsular region. Beginning on day 5, TUNEL-positive cells with morphologic characteristics of apoptosis and necrosis were detected in the adhesive region. Apoptotic cell death in this region was detected among the myofibroblast-like cells. Lens epithelial cells that had extended onto the central portion of the posterior capsule began to diminish on day 8 or 9, some of them morphologically demonstrating necrotic changes. CONCLUSIONS: Apoptosis of lens epithelial cells occurs in the process of wound healing and reepithelialization after lens extraction, leading to secondary cataract. If the induction of apoptosis is better understood, protocols might be developed that could prevent reepithelialization through apoptosis, thus delaying or preventing secondary cataracts. PMID- 9344356 TI - The initial complication rate of phacoemulsification in India. AB - PURPOSE: This study was designed to investigate the feasibility of teaching experienced surgeons to perform phacoemulsification in India, a cataract-endemic area. Complications occurring during surgery and the first postoperative day were documented and evaluated. METHODS: During a 1-month period, at the Aravind Eye Hospital in Madurai, India, the first 100 consecutive cataract operations performed by each of three experienced surgeons (a total of 300 cases), using phacoemulsification were prospectively evaluated. Multiple logistic regression was used to identify factors associated with intraoperative and postoperative complications. RESULTS: The mean age of patients was 57.4+/-9.3 years. The median best corrected preoperative visual acuity was 20/80. Mean surgical and phacoemulsification times were 15.8+/-3.7 minutes and 2.2+/-1.5 minutes, respectively. Complications occurred in 65 (21.7%) eyes. The most common was a rent in the posterior capsule, occurring in 40 (13.3%) eyes. There were significant variations in complication rate and in surgical time among the surgeons. The risk of experiencing a complication decreased as the number of phacoemulsifications performed increased. An increased risk of complications was associated with worse preoperative visual acuity and increasing patient age. CONCLUSIONS: With each successive case, the chances of experiencing a complication decreased 1%. Acceptable results were obtained within 1 month of performing the first phacoemulsification. PMID- 9344357 TI - Choroidal blood flow during isometric exercises. AB - PURPOSE: To investigate the response of choroidal blood flow in the foveal region of the human eye to increases in mean perfusion pressure (PPm = mean ophthalmic artery pressure - intraocular pressure; IOP) induced by isometric exercises. METHODS: Using laser-Doppler flowmetry, changes in velocity (ChBVel), number (ChBVol), and flux (ChBF) of red blood cells in the choroidal vascular system in the foveal region of the fundus were measured in both eyes of 11 normal subjects (ages 18 to 57 years) during isometric exercises. RESULTS: During 90 seconds of squatting, PPm increased by an average of 67%, from 46 to 77 mm Hg. This resulted in a significant increase of 12% in ChBFm (the mean of ChBF during the heart cycle), mainly caused by an increase in ChBVelm. A further increase in PPm to a value approximately 85% above baseline resulted in a 40% increase in ChBFm. A significant negative correlation was found between the changes in ChBVelm and ChBVolm, during squatting. CONCLUSIONS: Previous studies have demonstrated that during isometric exercise, blood pressures in the ophthalmic and brachial arteries rise in parallel. These observations and the current results indicate that an increase in PPm up to 67% induces an increase in choroidal vascular resistance that limits the increase in choroidal blood flow to approximately 12%. This regulatory process fails when PPm is further increased. PMID- 9344358 TI - Light exposure induces ubiquitin conjugation and degradation activities in the rat retina. AB - PURPOSE: To evaluate the consequences of light exposure on retinal ubiquitin (Ub) conjugation and degradation. METHODS: Two-month-old Long Evans pigmented rats were exposed to constant light (180 foot-candles) or were left in complete darkness for 18 hours. Rats used for cyclic light and diurnal rhythm experiments were removed from their light cycles at different times (24-hour clock): 0700 (before the light was turned on), 1000 (3 hours into the light cycle), 1000D (continued in the dark cycle), 1900 (before the light was turned off), 2200 (3 hours into the dark cycle), and 2200L (continued in the light cycle). The retinas were examined for Ub conjugation, adenosine triphosphate-Ub-dependent degradation, levels of Ub messenger RNA, and localization of Ub immunocytochemistry. RESULTS: There was a statistically significant increase in Ub conjugation and degradation in retinas isolated from light-exposed animals compared with degradation in retinas of dark-adapted animals. However, no significant differences were observed in the levels of Ub messenger RNA from cyclic light, or light-exposed or dark-adapted retinas, suggesting that light stress-induced changes do not reflect increased transcriptional activity. The daily variations observed in Ub conjugation and degradation suggest that these processes are probably the result of a circadian rhythm. Results of immunohistochemical studies revealed that Ub and its conjugates were uniformly distributed throughout the retinal cell layers in light- and dark-adapted retinas. However, in light-exposed retinas, a strong positive immunoreactivity was observed in the inner retina, specifically in horizontal and ganglion cells. CONCLUSIONS: These results suggest that light exposure may play a role in inducing Ub-conjugating activity in certain retinal cells. Furthermore, the results support the hypothesis that Ub is a stress protein that plays an important role in protecting cells under stress conditions. PMID- 9344359 TI - Evidence for photoreceptor changes in patients with diabetic retinopathy. AB - PURPOSE: To determine whether the rod and cone photoreceptors are affected in patients with diabetic retinopathy. METHODS: Twelve patients with diabetes and varying levels of retinopathy and nine age-similar control observers participated in this study. Two-color (500 versus 650 nm) dark-adapted thresholds were measured as a function of retinal eccentricity. Full-field flash electroretinograms were obtained using brief, high-intensity flashes. Dark adapted rod-isolated (Wratten 47B filter) and light-adapted cone-isolated (Wratten 26 filter) electroretinographic responses were measured as a function of flash intensity. The a-wave data were fitted with a model based on photopigment transduction to obtain values for the parameters of Rmax (the maximal response) and log S (sensitivity). Standard clinical 30-Hz flicker electroretinographic responses were also measured. RESULTS: Psychophysically measured dark-adapted thresholds were elevated primarily at eccentricities of 5 degrees and 10 degrees from the fovea. Analysis of rod and cone a-wave data showed that Rmax was normal in most of the patients, but log S was reduced. Analysis of b-wave and oscillatory potential parameters showed rod and cone postreceptoral abnormalities, including changes in the rod-isolated semisaturation constant (log k), cone-mediated 30-Hz flicker, and cone-isolated oscillatory potentials. The electrophysiological results were not significantly correlated with blood glucose or glycosylated hemoglobin level. CONCLUSIONS: The results provide evidence for rod and cone receptoral and postreceptoral deficits in patients with diabetic retinopathy. The photoreceptor changes are primarily in the log S (sensitivity) parameter and are attributed to transduction abnormalities. PMID- 9344360 TI - Identification of the retinal pigment epithelium protein RET-PE2 as CE-9/OX-47, a member of the immunoglobulin superfamily. AB - PURPOSE: To identify the retinal pigment epithelium (RPE) surface antigen recognized by the monoclonal antibody RET-PE2. METHODS: A lambda bacteriophage complementary DNA (cDNA) expression library, representing the rat RPE cell line RPE-J, was constructed and screened with the RET-PE2 monoclonal antibody. Transient transfections of the RET-PE2 cDNA, immunofluorescence stainings of tissue sections or cultured cells, and Western blot analyses of tissue and cell detergent extracts served to prove that the protein resulting from expression of the cDNA is the RET-PE2 antigen. RESULTS: Three independent cDNAs were cloned that shared overlapping sequences. Sequence alignment with EMBL database entries revealed identity to the published cDNA of CE-9/OX-47, a member of the immunoglobulin superfamily. One of the clones encoded the entire open reading frame of CE-9. The expression pattern of the RET-PE2 antigen matched that of CE 9, which is widely expressed. Chinese hamster ovary cells transiently transfected with the RET-PE2 cDNA produced a membrane-localized protein that was recognized by RET-PE2 and CE-9 antibodies. CONCLUSIONS: The antibody RET-PE2 recognizes the CE-9/OX47 gene product, a transmembrane protein of the immunoglobulin superfamily. Contrary to results reported earlier, RET-PE2 immunoreactivity is widely distributed among different rat tissues--kidney, liver, and testis. In epithelia other than the adult RPE, it is confined to the basolateral plasma membrane. Its apical polarization in the RPE of adult rats supports earlier findings that some proteins that are basolateral in other epithelia exhibit reversed polarity in the RPE. PMID- 9344361 TI - Immunochemical localization of the Batten disease (CLN3) protein in retina. AB - PURPOSE: Batten disease, also known as juvenile ceroid-lipofuscinosis and CLN3, is an autosomal recessively inherited disorder that results in blindness due to retinal degeneration. The CLN3 gene has been identified, but the function of the protein that this gene encodes is unknown. Experiments were conducted to determine where the CLN3 protein is localized in the mouse retina. Localization should provide a clue in evaluating potential functions of this protein. METHODS: Using oligonucleotide primers based on the reported human CLN3 cDNA sequence, the mouse cDNA nucleotide sequence was determined from products of the reverse transcriptase-polymerase chain reaction and 3' rapid amplification of cDNA ends. A synthetic 20-amino-acid peptide corresponding to an internal hydrophilic region of the predicted amino acid sequence of the mouse CLN3 protein was used to immunize rabbits. The resulting antiserum was used in immunoblot analysis of mouse retina homogenates and in electron microscopic immunocytochemical labeling of mouse retina sections. RESULTS: The peptide antibody labeled a single protein band of approximately 50 kDa on immunoblots of mouse retina homogenates. No labeling was detected with homogenates from human retinas. The antibody specifically labeled mitochondria of Muller cells and inner retinal neurons. Little labeling was observed in mitochondria of the photoreceptor cells. Mitochondria of other cell types, including the retinal pigment epithelium and choroidal cells, were not labeled. CONCLUSIONS: The retinal CLN3 protein appears to be localized almost exclusively in the mitochondria, but was detected only in certain cell types. Batten disease is characterized by massive lysosomal accumulations of a small inner mitochondrial membrane protein (subunit c of ATP synthase). The mitochondrial localization of the CLN3 protein suggests that it may play a role in the normal processing of subunit c. PMID- 9344362 TI - Diets enriched in docosahexaenoic acid fail to correct progressive rod-cone degeneration (prcd) phenotype. AB - PURPOSE: Results of a previous study show abnormal plasma lipids in progressive rod-cone degeneration (prcd)-affected dogs, with lower docosahexaenoic acid (DHA; 22:6n-3) and cholesterol levels but no differences in other plasma fatty acids, lipids, triglycerides, and fat-soluble vitamins. There is also an increase of the DHA precursor 22:5n-3, so that the ratio of 22:5n-3 to 22:6n-3 is higher in affected than in normal dogs. Because DHA is the predominant esterified fatty acid in rod outer segment (ROS) phospholipids, these findings suggest a possible causal association between abnormal plasma lipid levels and retinal degeneration. In the current study, dietary supplements rich in 22:6n-3 were used to determine whether plasma, liver, and rod outer segment phospholipid composition can be altered to modify the prcd disease phenotype. METHODS: prcd-affected and normal control dogs were given DHA-enriched supplements for short (7- and 25-day) and long (21-week) periods, and the fatty acid composition of plasma, liver, and rod outer segment phospholipids were examined. In the long-term study, electroretinography and morphology were used to assess modification of the retinal degeneration phenotype. RESULTS: Administration of DHA-enriched supplements resulted in increases in plasma DHA and n-3 polyunsaturated fatty acids and in decreases in some n-6 fatty acids in normal and prcd-affected dogs. Similar increases in DHA and n-3 fatty acids were observed in the liver, but affected dogs had significantly higher levels at all supplementation time points examined. In contrast, the ROS of affected dogs had statistically lower (approximately 20%) DHA levels, and these levels could not be increased with dietary supplementation. The disease phenotype could not be modified by DHA enriched supplements. CONCLUSIONS: Regardless of the sustained three- to fourfold elevation in plasma and liver DHA that occurs as the result of supplementation, the ROS DHA levels remain unchanged, and the prcd disease phenotype is not modified by the dietary manipulation. These findings could be the result of a reduction in the synthesis of DHA-containing phospholipids in the retinas of affected dogs; or, alternatively, there could be a reduction in DHA uptake, transport, or storage within the retinal pigment epithelium-photoreceptor complex. PMID- 9344363 TI - Type XII collagen contributes to diversities in human corneal and limbal extracellular matrices. AB - PURPOSE: To characterize diversities in the extracelhtlar matrices (ECMs) of the corneal and the surrounding limbal epithelium and stroma. METHODS: Immunohistochemical analyses were employed for screening monoclonal antibodies (mAbs) developed against ECM components of the human corneal epithelial basement membrane (BM) zone. In the current study, mAb BM8 was used as the monospecific probe to characterize its antigen (AgBM8) immunochemically, and to immunoselect a complementary DNA (cDNA) clone encoding AgBM8. Direct biochemical and cDNA sequence analyses were performed for the further characterization of AgBM8. An indirect colloidal gold-conjugated antibody technique was employed for immunoelectron microscopic analysis to study the distribution of AgBM8 in the corneal ECMs. RESULTS: The protein AgBM8, isolated from rabbit corneal stromal and epithelial tissues, was identified as the long-splice variant form of type XII collagen based on its size (approximately 340 kDa disulfide-linked subunits), the presence of collagenous domain(s) and a noncollagenous domain of approximately 300 kDa in its subunit structure, and its internal amino acid sequences. The identity of AgBM8 was further confirmed from the amino acid sequence (517 amino acids) deduced from the sequence of a cDNA immunoselected with mAb BM8. Immunofluorescence analyses indicated that the long form of type XII collagen is present in the ECMs of corneal stroma and in the sclera, as well as in the corneal epithelial BM zone but is absent in the limbal and conjunctival epithelial BM zones. It was not detectable in the subepithelial loose connective tissues in the limbus and in the bulbar conjunctiva. Immunoelectron microscopic analyses indicated that the long variant form of type XII collagen is present in corneal epithelial BM, Bowman's membrane, and the interfibrillar matrix of the corneal stroma. In the stroma, colloidal gold was distributed along the collagen fibrils with a periodicity of 150 to 200 nm. CONCLUSIONS: The long variant form of human type XII collagen, a member of the fibril-associated collagens with interrupted triple helices, referred to as FACITs, contributes to the differences in the BM zones of the cornea and limbus. Although many of the dense connective tissues in adult animals contain the short variant form of type XII collagen, human corneal stroma, the BM zone, and the sclera contain the long variant form as the predominant form of type XII collagen. In the corneal stroma, type XII collagen may be organized along the collagen fibrils in a uniform head-to-tail pattern. PMID- 9344364 TI - Astrocytes increase barrier properties and ZO-1 expression in retinal vascular endothelial cells. AB - PURPOSE: Diabetic retinopathy and other diseases associated with retinal edema are characterized by increased microvascular leakage. Astrocytes have been proposed to maintain endothelial function in the brain, suggesting that glial impairment may underlie the development of retinal edema. The purpose of this study was to test the effects of astrocytes on barrier properties in retinal microvascular endothelial cells. METHODS: Bovine retinal microvascular endothelial cells were exposed to conditioned media from rat brain astrocytes. Transendothelial electrical resistance (TER) was determined on 24-mm Transwell (Cambridge, MA) polycarbonate filters with the End-Ohm device (World Precision Instruments, Sarasota, FL). ZO-1 protein content was quantified by microtiter enzyme-linked immunosorbent assay. RESULTS: Astrocyte-conditioned medium (ACM) significantly increased TER (P < 0.0001) and ZO-1 content (P < 0.01). Both serum containing and serum-free N1B defined ACM increased ZO-1 expression, but heating abolished the effect. Serum-free ACM decreased cell proliferation by 16%. CONCLUSIONS: Astrocytes release soluble, heat-labile factors that increase barrier properties and tight junction protein content. These results suggest that astrocytes enhance blood-retinal barrier properties, at least in part by increasing tight junction protein expression. Our findings suggest that glial malfunction plays an important role in the pathogenesis of vasogenic retinal edema. PMID- 9344365 TI - Suppression of nuclear factor kappa B and CD18-mediated leukocyte adhesion to the corneal endothelium by dexamethasone. AB - PURPOSE: To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFkappaB). METHODS: Cultured bovine corneal endothelium was stimulated for 6 hours by 40 micron/ml tumor necrosis factor alpha (TNFalpha). Dexamethasone was added 1 hour before TNFalpha stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37 degrees C, and then neutrophils were activated by N-formyl-methionyl-leucyl phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkappaB after TNFalpha simulation was determined by confocal immunocytochemistry using an anti p65 antibody. RESULTS: Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFalpha (451.4+/-45.4 cells/mm2, n = 16) compared to control (156.7+/-27.3 cells/mm2, n = 16, P < 0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6+/-25.1 cells/mm2, n = 8, P < 0.01) and by pretreatment with 10(-7) M dexamethasone (207.9+/-31.5 cells/mm2, n = 10, P < 0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFkappaB was located in the cytoplasm in unstimulated cells; however, the addition of TNFalpha caused NFkappaB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFkappaB translocation into the nucleus. CONCLUSIONS: Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFkappaB entry into the nucleus. PMID- 9344366 TI - Treatment of major depression: selection of initial drug. PMID- 9344367 TI - Declaration of treatment failures. PMID- 9344368 TI - Augmentation strategies for treatment of unipolar major depression. PMID- 9344369 TI - Switching strategies for the treatment of unipolar major depression. PMID- 9344370 TI - Selection of the initial drug(s) in the treatment of bipolar disorder, depressed phase. PMID- 9344371 TI - Treatment of bipolar disorder, depressed phase augmentation/switching strategies. PMID- 9344372 TI - Selection of initial treatment for bipolar disorder, manic phase. PMID- 9344374 TI - The psychopharmacological treatment of nonmajor mood disorders. PMID- 9344373 TI - Algorithms for bipolar mania. PMID- 9344375 TI - Depression in patients with somatic diseases. PMID- 9344376 TI - Treatment of depression in the elderly. PMID- 9344377 TI - The role of the patient in treatment decisions. PMID- 9344378 TI - The decision to use ECT: for whom? When? PMID- 9344379 TI - Light therapy for depressive disorders: indications and efficacy. PMID- 9344380 TI - Special treatment issues: maintaining and discontinuing psychotropic medications. PMID- 9344400 TI - The role of estrogen in the treatment and prevention of dementia: introduction. PMID- 9344401 TI - Current concepts in the pathogenesis of Alzheimer's disease. AB - Alzheimer's disease (AD) affects a large proportion of the increasingly aging population of this country, with prevalence rates as high as 47% for those >85 years old and a total annual cost approaching $70 billion. There is no currently validated test for detection of dementia of the Alzheimer type (DAT). Because of this and the insidious onset of the disease, the diagnosis may be missed by primary care physicians. Cerebral extracellular beta-amyloid deposition as senile plaques and intraneuronal neurofibrillary tangles appear to represent critical processes in the development of AD; however, whether and the extent to which these may also occur in nondemented aging is uncertain. Tangles occur primarily in medial temporal lobe structures (hippocampus, entorhinal cortex, and amygdala), and tangle density correlates with dementia severity. Plaques are diffusely distributed throughout the cerebral cortex, and are the neuropathologic hallmark of the disease. Aging is the primary risk factor for AD. After controlling for differential life expectancy, female sex still appears to be an additional risk factor. There may be a genetic component, in some cases based on family and twin studies. Allelic variation in the apolipoprotein E (Apo E) gene located on chromosome 19 represents another important risk factor. However, the diversity of gene mutations apparently responsible for the various forms of AD suggest that the disease is genetically heterogeneous. AD may be conceptualized as an imbalance between neuronal injury and repair. Oxygen free radicals may be involved in the cross-linking process of beta-amyloid aggregation, and antioxidants may represent a potential intervention. There may be a role for heavy metals in the pathogenesis of AD, but this remains controversial. Work continues toward possibly a cure or prevention, but more likely palliation, of AD, and the results of trials of anti-inflammatory agents, estrogen, and antioxidant therapy are anticipated in the near future. PMID- 9344402 TI - Estrogen, cognition, and a woman's risk of Alzheimer's disease. AB - Alzheimer's disease affects women more often than men, and women with this form of dementia show greater naming (semantic memory) deficits during the course of their illness. Gonadal steroids exert organizational and activational effects on central nervous system neurons and influence brain function in other important ways. Several estrogenic actions are potentially relevant to Alzheimer's disease, and it is hypothesized that one consequence of estrogen deprivation after the menopause is a higher risk of this dementing disorder. In healthy women without dementia, estrogen may enhance cognitive performance, especially in the domain of verbal memory, although the magnitude of such effects is small. Several small treatment trials of estrogen replacement in women with Alzheimer's disease, however, suggest that estrogen's effects on cognition could be larger in this population and may be most apparent on tasks of semantic memory. Analyses in voluntary cohorts associate postmenopausal estrogen replacement therapy with a lower risk of subsequent Alzheimer's disease. In 3 recent epidemiologic studies, information on postmenopausal estrogen use was collected prospectively; while inconclusive, findings raise the possibility that postmenopausal estrogen replacement reduces a woman's risk of subsequent dementia. New information from basic research and from large randomized treatment studies, cohort studies, and case-control studies is needed to resolve important unanswered clinical issues. PMID- 9344404 TI - Estrogen replacement therapy and cognitive function in postmenopausal women without dementia. AB - The epidemiologic evidence for an association between estrogen and cognitive function among healthy postmenopausal women remains controversial. Equivocal findings may be explained, in part, by differences in the methodologic approaches of these studies. Overall, the evidence for a positive relationship comes primarily from randomized clinical trials. These trials suggest an acute effect on specific tests of recent verbal memory and tasks incorporating concept formation and reasoning. The potential long-term effects of estrogen in slowing or delaying the age-related decline in cognitive function require further study. More data are needed to determine the effects of estrogen replacement therapy on cognitive function, independent of changes in mood and depressive symptoms. In addition, evidence suggests that progesterone may mitigate the beneficial effects of estrogen on mood. Research should be undertaken to determine the interactive effects of estrogen and progesterone on cognitive function. Lastly, there should be continued investigation by both epidemiologic and basic neuroscientific studies to further elucidate the specific cognitive domains that may respond to estrogen. PMID- 9344403 TI - Role of estrogen replacement therapy in memory enhancement and the prevention of neuronal loss associated with Alzheimer's disease. AB - Recent evidence supports a role for estrogens in both normal neural development and neuronal maintenance throughout life. Women spend 25-33% of their life in an estrogen-deprived state and retrospective studies have shown an inverse correlation between dose and duration of estrogen replacement therapy (ERT) and incidence of Alzheimer's disease (AD), suggesting a role for estrogen in the prevention and/or treatment of neurodegenerative diseases. To explore these observations further, an animal model was developed using ovariectomy (OVX) and ovariectomy with estradiol replacement (E2) in female Sprague-Dawley rats to mimic postmenopausal changes. Using an active-avoidance paradigm and a spatial memory task, the effects of estrogen deprivation were tested on memory-related behaviors. OVX caused a decline in avoidance behavior, and estrogen replacement normalized the response. In the Morris water task of spatial memory, OVX animals showed normal spatial learning but were deficient in spatial memory, an effect that was prevented by estrogen treatment. Together these data indicate that OVX in rats results in an estrogen-reversible impairment of learning/memory behavior. Because a plethora of information has been generated that links decline in memory related behavior to dysfunction of cholinergic neurons, the effects of estrogens on cholinergic neurons were tested. We demonstrated that OVX causes a decrease in high affinity choline uptake and choline acetyltransferase activity in the hippocampus and frontal cortex; ERT reverses this effect. Further, we showed that estrogens promote the expression of mRNA for brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), 2 neurotrophic substances that have been shown to ameliorate the effects of age and injury on cholinergic neurons. Tissue culture models were used to evaluate whether estrogen treatment increases the survival of neurons when exposed to a variety of insults. 17-beta-Estradiol (beta E2) protects cells from the neurotoxic effects of serum deprivation and hypoglycemia in human neuroblastoma cell lines. We have also observed that 17 alpha-estradiol (alpha-E2), a weak estrogen, shows neuroprotective efficacy in the SK-N-SH cell line at concentrations equivalent to beta-E2. Finally, we have observed that tamoxifen, a classic estrogen antagonist, blocks only one-third of the neuroprotective effects of either alpha-E2 or beta-E2. Collectively, these results indicate that estrogen is behaviorally active in tests of learning/ memory; activates basal forebrain cholinergic neurons and neurotrophin expression; and is neuroprotective for human neuronal cultures. We conclude that estrogen may be a useful therapy for AD and other neurodegenerative diseases. PMID- 9344405 TI - Estrogen and the treatment of Alzheimer's disease. PMID- 9344406 TI - Potential role for estrogen replacement in the treatment of Alzheimer's dementia. AB - In light of evidence that estrogen replacement therapy (ERT) might affect cholinergic function, we examined possible effects of ERT on clinical and cognitive responses to the cholinesterase inhibitor tacrine in women with Alzheimer's disease (AD). In a previously reported 30-week, randomized, double blind, placebo-controlled, multicenter clinical trial, 14.5% of 318 women with evaluable data had been receiving ERT prior to randomization. Patients were randomly assigned to receive placebo or one of three ascending dosages of tacrine (maximum dosages of 80 mg/day, 120 mg/day, or 160 mg/day). Women completing the trial receiving ERT and tacrine improved more than women not receiving ERT who were randomized to tacrine or to placebo as assessed by cognitive (p <0.01), clinical (p = 0.02), caregiver (p = 0.006), and mental status (p = 0.07) ratings. Using an intent-to-treat analysis, they improved significantly on cognitive ratings (p = 0.01). These results provide evidence that prior and continuing ERT may enhance response to tacrine in women with AD. Randomized trials are needed. PMID- 9344407 TI - BIACORE analysis of histidine-tagged proteins using a chelating NTA sensor chip. AB - While BIACORE instruments are routinely used for kinetic measurements and for the determination of binding constants, the immobilization of a ligand onto the sensor chip surface has to be individually optimized for every system. We show here that the histidine (His) tag, routinely used in protein purification and in detection is an ideal tag for immobilization, despite the intrinsically low affinity between an immobilized metal ion and the His tag. This is due to strong rebinding effects caused by the high surface density of immobilized Ni2+ nitrilotriacetic acid (NTA) on the chips used here. The immobilization of the ligand can be adjusted to a low level using the same chip, such that mass transport limitation and rebinding of the analyte to the immobilized ligand is minimal. Nine different proteins with different numbers of His tags were tested for stable binding to the Ni2+-NTA surface. Most proteins with one His tag dissociate very rapidly from the Ni2+-NTA surface, and the KD for the interaction between His tag and Ni2+-NTA was estimated to about 10(-6) m at neutral pH. In contrast, two His tags are usually found to be sufficient for stable binding. The kinetics of the chaperonin system of Escherichia coli GroEL and GroES were analyzed as a model using this system and found to be very similar to those obtained with covalently immobilized ligands. The sensor chip can be reused many times, because of the powerful regeneration methods. The ligand can be freshly immobilized after each cycle, thus eliminating potential denaturation upon regeneration as a source of error. PMID- 9344408 TI - Rat liver theta-class glutathione S-transferases T1-1 and T2-2: their chromatographic, electrophoretic, immunochemical, and functional properties. AB - A method was established for simultaneously isolating Theta-class glutathione (GSH) S-transferases (GSTs) T1-1 and T2-2 as homogeneous proteins from rat (r) liver cytosol. The established method of using an 8-aminooctyl Sepharose 4B column to separate rGSTT1-1 from rGSTT2-2 at the final stage of their purification was a modification of the method previously reported for the isolation of rGSTT2-2 (Hiratsuka et al., J. Biol. Chem., 265, 11973-11981, 1990). Specific substrates used for purification of the Theta-class rGSTs were dichloromethane for T1-1 and 5-sulfoxymethylchrysene for T2-2. rGSTsT1-1 and T2-2 existed at a ratio of 1:7 at a total concentration of 0.5% of that of the cytosolic protein. Purified rGSTsT1-1 and T2-2 were separated as single bands at 28 and 26.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and as single peaks at retention times of 36 and 34 min, respectively, by reverse phase partition high-performance liquid chromatography on a microBondasphere column eluted with a linear gradient of acetonitrile in water containing trifluoroacetic acid. Western blot analysis indicated that rabbit antisera raised against rGSTsT1-1 and T2-2 intensely reacted with the corresponding antigens, but showed no detectable reactivity with the different isoforms of Theta-class rGSTs as well as with representative hepatic rGSTs of other classes. The Theta-class rGSTs showed higher GSH peroxidase activity than rGSTA1-2 toward hydroperoxides of cumene, arachidonic acid, and linoleic acid. Cumene hydroperoxide was a better substrate for rGST T1-1 than for rGST T2-2, while the fatty acid hydroperoxides were the better substrates for rGST T2-2 than for rGST T1-1. PMID- 9344409 TI - A continuous colorimetric assay for rhinovirus-14 3C protease using peptide p nitroanilides as substrates. AB - Human rhinovirus encoded 3C protease is an attractive target for antiviral drug development. However, lack of a convenient and selective assay for 3C protease has been a hindrance in characterization of this enzyme and evaluation of a large number of potential inhibitors. In the present study we describe development of a simple, continuous colorimetric assay for this enzyme using peptide p nitroanilides (pNA) as substrates. Several peptides mimicking the native 3C cleavage site of HRV-14 polyprotein have been synthesized with an N-acylated p nitroaniline at position P1' and examined as substrates for the purified 3C protease. In these peptides, amino acids downstream from the original cleavage site have all been replaced with a chromophoric p-nitroaniline moiety which is directly linked to the bond undergoing enzymatic cleavage, thereby generating a new cleavage site Gln-pNA for the enzyme. Hydrolysis of these pNA peptides by 3C at the newly formed scissile bond releases free p-nitroaniline which is yellow colored and can be continuously monitored at a visible wavelength. Kinetic parameters of 3C protease toward these peptides have been measured and analyzed. In addition, the pNA peptides have been modeled within the active site of the 3C protease to investigate the ability of the pNA group to act as a replacement for Gly-Pro in the prime side. The selectivity and applicability of this assay and its advantages over the previously described methods have been demonstrated and discussed. Since multiple tests can be performed simultaneously in one microtiter plate, the assay is ideal for evaluation of a large number of samples. PMID- 9344410 TI - Characterization of single-cell migration using a computer-aided fluorescence time-lapse videomicroscopy system. AB - Single-cell assays of cell migration, while yielding dynamic measurements of cell position and morphology, are predominantly limited by the time required for data collection and analysis. Computer-aided fluorescence time-lapse videomicroscopy (CAFTiV) was developed in order to facilitate the tracking and rapid examination of large numbers of motile cells. The system combines time-lapse videomicroscopy with epifluorescence capability, which allows full automation of image capture, sorting, and analysis due to the low background in the fluorescence images. Utilizing the CAFTiV system, data analysis time was reduced from over 125 h to less than 1 labor minute. In addition, fluorescence imaging permits cell tracking in small-volume chambers (<100 microL), which is useful should the addition of expensive reagents be required. It is anticipated that the ability to characterize both biochemical and biophysical properties responsible for cell movement will be enhanced by this methodology. PMID- 9344411 TI - Measurement of histidinohydroxylysinonorleucine and hydroxyproline in skin collagen by reversed-phase high-performance liquid chromatography after 9 fluorenylmethyl chloroformate labeling. AB - A novel, highly sensitive method to quantify histidinohydroxylysinonorleucine (HHL), a trifunctional type of cross-link in skin collagen, was developed. HHL in skin hydrolysates labeled with 9-fluorenylmethyl chloroformate (FMOC-Cl) was separated by reversed-phase high-performance liquid chromatography. Mass spectrometric analysis revealed that two FMOCs were bound to two primary amino acid residues, histidine and hydroxylysine, but not to lysine residue in one HHL molecule. Hydroxyproline was simultaneously measured to express the molar ratio of HHL to collagen. The detection range of HHL was from 1 to 10 pmol and that of hydroxyproline from 1 to 50 pmol. A 6-mm punch-biopsied human skin sample contained 0.40 to 0.69 mol of HHL per one molecule of collagen. This sensitive method is useful as it is rapid and can be used to examine the aging process or the change of HHL content in skin collagens of various pathologic states. PMID- 9344412 TI - Preparation of enantiomerically pure L-7-azatryptophan by an enzymatic method and its application to the development of a fluorimetric activity assay for tryptophanyl-tRNA synthetase. AB - The reaction of D,L-7-azatryptophan (D,L-7AW) with tryptophanyl-tRNA synthetase (TrpRS), adenosine triphosphate (ATP), and Mg2+ in the presence of inorganic pyrophosphatase results in the formation of a highly fluorescent l-7AW-adenylate complex. Detection of this complex is based on its enhanced fluorescence at 315 nm excitation and 360 nm emission after the addition of ATP. This stereoselective reaction was used to develop an activity assay for TrpRS using commercially available racemic D,L-7AW. The assay can be used to determine the activity of TrpRS from samples which contain less than 1 nmol of enzyme in 250 microL of sample. Thus the enzyme activity can be assessed without resorting to a radioactive assay of tRNATrp acylation. A secondary use of the stereoselective assay was for confirming the presence of pure L-7AW, D-7AW, or mixtures of the two enantiomers. D-7AW and L-7AW were prepared by reacting D,L-7AW with chloroacetic anhydride to form N-chloroacetyl-D,L-7AW (ClAc-7AW) followed by stereospecific proteolytic digestion of ClAc-7AW using carboxypeptidase A to produce the free L-7AW. The L-7AW could be separated from unreacted N chloroacetyl-7AW by reverse-phase HPLC. The TrpRS-based assay was able to unambiguously discriminate between the two enantiomers of 7AW. The assay was then used to identify which enantiomer of 7AW was present in resolved fractions of the tripeptide L-lysyl-D,L-7-azatryptophyl-L-lysine. Digestion of the resolved tripeptides with protease enzymes produced the free L or D enantiomer of 7AW, which was easily identified using the TrpRS assay procedure. PMID- 9344413 TI - Perils of partitioning: A case study of flavins and flavokinase. AB - Partitioning is a common procedure for the separation of two solutes from a solution based on their differential solubility in an immiscible solvent. This has been widely used to quantitate riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide in aqueous samples by extraction with water-saturated benzyl alcohol. Here we report that the partitioning of riboflavin and FMN is affected by the presence of each other in a concentration-dependent manner, thus rendering this procedure unsuitable for quantitation. Direct quantitation of FMN formed in assays for flavokinase from Vigna radiata shows that kinetic analyses using a partition-based assay lead to erroneous conclusions. PMID- 9344414 TI - Inhibition, reactivation, and determination of metal ions in membrane metalloproteases of bacterial origin using high-performance liquid chromatography coupled on-line with inductively coupled plasma mass spectrometry. AB - High-performance liquid chromatography coupled on-line with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was used for the characterization of metal ions in several metalloproteases of bacterial origin. The different components of the bacterial extracts were separated on a size-exclusion column. The eluent of the HPLC system was continuously transported to the ICP-MS system for rapid, reproducible, and sensitive analyses of trace elements in the metalloproteases. Two different membrane proteases from Bacillus cereus and Pseudomonas aeruginosa were characterized to be zinc metalloproteases using enzymological methods and HPLC-ICP-MS. The zinc content was determined to be three molecules of zinc per protein molecule for the B. cereus protease and one molecule of zinc per protein molecule for the P. aeruginosa protease. For another purified protease, a periplasmic alanyl aminopeptidase of P. aeruginosa, the lack of protein-bound metal ions could be clearly determined-a confirmation that this main aminopeptidase of P. aeruginosa belongs to the cysteine protease family. The presence of nonionic detergents can influence the distribution of trace elements during the HPLC separation. Therefore, the use of these substances should be avoided during enzyme purification for metal analyses or they should be exchanged later for zwitterionic and ionic detergents with more strongly dissociating properties. PMID- 9344415 TI - An autocatalytic expression system for regulated production of recombinant protein in mammalian cells. AB - An autocatalytic inducible mammalian expression system was established. This system is composed of two sets of vectors: one carries a selectable marker gene and the other carries a bicistronic expression unit consisting of a target gene, an internal ribosomal entry site, and a tetracycline-controlled transactivator gene. Both the selectable marker and the bicistronic unit are controlled by the tetracycline-responsive promoter (PhCMV*-1). When the two vectors are cotransfected into host cells, only a single selection round in the absence of tetracycline is necessary to generate clones expressing the target gene. The expression level is high and easily regulated by tetracycline. Combination with a gene amplification system may allow further enhance the foreign gene expression. Because the PhCMV*-1 promoter is relatively independent of cellular regulation signals, this system is expected to work in a wide variety of cell types. PMID- 9344416 TI - Determination of pheomelanin by measurement of aminohydroxyphenylalanine isomers with high-performance liquid chromatography. AB - We describe an improved method for the analysis of pheomelanin in biological samples. The method is based on a chemical degradation of the melanin polymer and HPLC analysis of specific degradation products. Hydriodic hydrolysis provides 4 amino-3-hydroxyphenylalanine (AHP) and 3-amino-l-tyrosine (AT) which are detected with an electrochemical detector. We have examined each step of the analysis and the results are presented in this paper. First the samples are hydrolyzed for 16 h. AT and AHP are then isolated from the hydrolysates by ion-exchange chromatography and then separated and quantitated by HPLC and electrochemical detection. The method shows good reproducibility with a total imprecision below 5.6%. The linearity of the method was shown from 0 to 490 ng AT and 0 to 850 ng AHP per sample, using a melanoma cell suspension (27 mg protein/ml) with up to 24 fold dilutions of the original sample. For cultured "normal" human melanocytes a minimal amount of 0.1 mg protein is sufficient for analysis of pheomelanin in the samples. This method provides the opportunity to study the composition of the formed melanin in cell lines, cultured in different growth media. PMID- 9344418 TI - A reference method for the analysis of aldosterone in blood by high-performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry. AB - A high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (HPLC-APCI-MS/MS) reference method for the quantitation of aldosterone in serum and plasma has been developed. Samples were extracted with dichloromethane/diethyl ether, containing flumethasone as internal standard (IS). Chromatography was performed on a phenyl column using 50 mm ammonium formate (pH 7.1)/methanol (50/50, v/v) as mobile phase. Analysis was in negative ionization mode by selected reaction monitoring (aldosterone m/z 359.2 --> 331.2; IS m/z 455.0 --> 379.0). The assay was linear over the range 15-500 pg/mL, with limits of detection and quantitation of 10 and 15 pg/mL, respectively. Imprecisions of the assay at 15, 20, 150, and 450 pg/mL were 18.5, 8. 8, 10.6, and 9.5%, respectively. The accuracy of the method ranged from 93.1 to 98.9% with absolute recoveries between 84.0 and 91.3% (aldosterone) and 88.0 and 92.3% (IS). We present a case study of a patient admitted, with suspected primary hyperaldosteronism, on the basis of a high radioimmunoassay (RIA) aldosterone concentration. The results suggest that RIA was unreliable, causing unnecessary patient discomfort and a costly 6-day hospital stay. The specific HPLC-API-MS/MS assay described offers the sensitivity and accuracy required to assess abnormal aldosterone production in hypertensive patients. PMID- 9344417 TI - A protease-free assay for peptidyl prolyl cis/trans isomerases using standard peptide substrates. AB - Peptidyl prolyl cis/trans isomerases (PPIases) are ubiquitous and abundant enzymes catalyzing peptide bond cis/trans isomerization adjacent to proline in peptides and proteins. An uncoupled protease-free assay of PPIase activity has been developed using the standard tetrapeptide substrates of the proteolytically coupled test system. Differences in the UV/vis absorption spectra of cis and trans conformations of Suc-Ala-Xaa-Pro-Phe-(Y-) anilide (Xaa = Ala, Leu, Phe; Y = 4-nitro, 2,4-difluoro) were exploited to monitor the time course of the cis/trans isomerization subsequent to a solvent jump from 0.47 M LiCl/trifluoroethanol into aqueous solution. The utility of the assay has been demonstrated by the determination of the Michaelis-Menten constants of cytosolic cyclophilin (Cyp18) and of the proteolytically sensitive FK506-binding protein-like PPIase SlyD from Escherichia coli. Furthermore, similar inhibition constants were estimated for the reversible inhibition of human Cyp18 by cyclosporin A (CsA) with both the proteolytically coupled and the novel uncoupled PPIase assay. PMID- 9344420 TI - Introduction to papers presented at: the twenty fifth anniversary symposium of the Columbia University Seminar on Appetitive Behavior. PMID- 9344419 TI - Mapping of recombinant hemoglobin using immobilized trypsin cartridges. AB - A tryptic mapping procedure has been developed for a recombinant hemoglobin (rHb1.1) using an immobilized trypsin cartridge. Apohemoglobin is passed through the trypsin cartridge and the products of the digestion are captured directly onto an in-line C18 reversed-phase column. The peptides are then separated using a gradient elution. This new procedure is rapid and reproducible and can be fully automated. The total time of analysis is less than 2 h. The mapping of apohemoglobins produced an unexpected isomerization of two peptides: beta8,9 (K66 K82) and alpha8,9 (K61-K90). It appears that the isomerization may occur through transpeptidation followed by proteolysis at a newly generated site next to the site of ligation. This mapping procedure can be a useful tool for research and routine analysis of proteins. PMID- 9344421 TI - The coming of genetics in the control of ingestion. AB - The study of appetitive behavior in relation to obesity is now being enriched by molecular-genetic findings in experimental animals with genetic obesity and associated "feeding disorders". The opinion is expressed that the mutual enrichment of the disciplines of behavior and molecular biology will place the study of human ingestive behavior on a firm scientific base. PMID- 9344422 TI - Neuroscience and appetitive behavior research: 25 years. AB - Neuroscience techniques have made major contributions to the understanding of appetitive behavior. Highlights in six areas are summarized to illustrate progress during the 25 years of the Columbia Appetitive Behavior Seminar: (1) discovery of angiotensin and aldosterone in the control of thirst and salt appetite; (2) electrophysiological decoding of chemoreceptive information in the brain; (3) a new foundation in the hypothalamus built on peptides, such as neuropeptide Y and galanin, interacting with monoamines and steroids in the control of appetite for macronutrients; (4) discovery of numerous peptides that mediate and integrate satiety, such as cholecystokinin, insulin, leptin and enterostatin, and other systems that suppress eating during illness; (5) better understanding of appetite suppressant drugs, and (6) exploration of a circuit that translates hypothalamic signals into behavioral action through connections to brainstem reflex arcs and forebrain instrumental response systems. PMID- 9344424 TI - Learned controls of ingestive behaviour. AB - During the 25 year history of the Columbia Appetitive Seminar there have been many notable developments in ingestive behavior research. One area of rapid progress concerns learned controls of feeding behavior. During the 1960's and 1970's most research related to food learning focused on conditioned flavor aversions. While it was assumed that animals also learned to prefer foods based on their positive nutritive consequences, there were few experimental demonstrations of this effect. Examples often cited involved animals learning to prefer a flavor associated with recovery from illness or a vitamin deficiency. There were isolated reports in the 1960's of nutrient infusions increasing flavor preference and acceptance, but it wasn't until the 1970's that nutrient-based learning was firmly established, and not until the 1980's and 1990's investigated in detail. This brief review highlights some of the major findings of nutrient based learning. Other important aspects of food learning (social, cultural, ecological, environmental) are not discussed here. PMID- 9344423 TI - Metabolic and hormonal controls of food intake: highlights of the last 25 years- 1972-1997. AB - The six major research advances in metabolic and hormonal controls of food intake that have altered the direction or have broadened the scope of the field in the last 25 years are discussed. The advances selected are: (1) GI processes and meal termination-the CCK pathway; (2) Brain insulin hypothesis; (3) Glucose-dependent processes in periphery, plasma, and brain including the transient declines in blood glucose signaling meal initiation; (4) Fatty acid oxidation in the liver; (5) Behavioral and metabolic patterns; and (6) New pathways from molecular genetics and molecular biology-the OB protein pathway. PMID- 9344425 TI - Who is in charge? Animal vs experimenter control. AB - In the 1920s Curt Richter (1927) stated that the central problem for psychology was to discover the determinants of the initiation and termination of bouts of behavior. Ignoring this challenge, experimentation in animal psychology has been dominated by the session paradigm in which animals work in brief sessions for a resource of which they have been deprived. In this open economy, no behavioral strategy of the animal can meet its demand, and the beginnings and ends of bouts are controlled by the experimenter; thus, Richter's problem cannot be addressed. In contrast, in a free-feeding, closed economy, the animal controls the initiation and termination of feeding and can regulate its intake, and bout patterns can be observed. If the paradigm is modified to simulate a habitat where resources are distributed discontinuously and the animal must work to discover and procure access to a commodity before it can be used, behavioral strategies allowing the animal to regulate its intake while tending to maximize the ratio of benefits to costs are revealed. We offer an answer to Richter's question based on a cost/benefit analysis of feeding behavior in this foraging paradigm. We show that the time and energy costs of resource acquisition and resource consumption are powerful determinants of the pattern of resource use, and that they have different and independent effects. The former costs are reduced by reducing the frequency of initiating bouts, and the latter costs, by altering the rate and amount of consumption. Further, the time window of these relations is much longer than expected from analyses in the session paradigm. We conclude that the recurrent nature of behavior is due to the discontinuous distribution of resources rather than to cycles of physiological depletion and repletion, and that the determinants of bout initiation and termination lie in the economics of the allocation of time and effort to different resources and activities. PMID- 9344426 TI - Eating disorders: the last 25 years. AB - During the past 25 years there has been measurable progress in the understanding and treatment of the three eating disorders-anorexia nervosa, bulimia nervosa and binge eating disorder. This progress has occurred in four areas-diagnosis, measurement, etiology and treatment. The introduction of diagnosis into the field of eating disorders, which involved a new emphasis on measurement, facilitated studies of epidemiology, etiology and treatment of what have become discrete clinical disorders. The greatest progress has been with bulimia nervosa, in which new behavioral and pharmacological treatments have reversed the pessimism with which the treatment of this disorder had initially been viewed. PMID- 9344427 TI - Eating and the American Zeitgeist. AB - This article traces the development of the science of eating in the United States from the experiment of Cannon and Washburn in 1912 until the founding of the Appetitive Seminar at Columbia in 1972. The interplay between the intrinsic aspects of science that depend on the sequence of ideas, techniques, and results, and the extrinsic influences of the Zeitgeist is emphasized. PMID- 9344428 TI - Food intake and meal patterns of one year old infants. AB - Nutrient intakes and meal patterns of 8.6 to 15.6 month old infants were investigated by analysing data collected by the caregivers of 29 infants. The caregivers maintained a 7-day diary which included everything the infant ate and other factors including people present during meal time and time of meals. Daily intake and meal size increased as age increased for the infants. A high level of variability in meal size was found, however, the variability in daily intake was much lower indicating an ability to adjust intake at meals to maintain a relatively stable daily intake. Infants were found to be responsive to their stomach contents, however, the circadian rhythm of intake was absent and social facilitation of intake was blunted. It is suggested that these differences are due to the fact the infants have not yet learned to respond to the social and environmental factors which markedly influence the intake of adults. PMID- 9344429 TI - Habituation of facial muscle responses to repeated food stimuli. AB - We have shown in a series of studies that the human salivary response habituates to repeated presentation of gustatory cues. Parallel animal research has shown that mouthing and food acceptance also habituate. Facial expressions represent a complex response pattern in animals and humans that may provide an objective measurement of motivation to eat. The current study assessed whether facial muscles (orbicularis oris region, risorius region, and zygomaticus region) that regulate mouthing habituate to repeated presentations of a small amount (3 kcal) of a pleasant tasting food stimulus. Participants were randomly assigned to groups that received ten presentations of the same or changing taste stimuli. Following these trials all subjects were presented a novel dishabituating stimulus followed by the habituating stimulus. Integrated facial electromyography EMG responses over each region were measured for a 50 second period following the taste stimulus. Results showed differential habituation rates across the two groups, with participants in the repeated taste group showing decreases in contrast to the changing taste group who did not decrease. The participants in the repeated taste group showed dishabituation of these responses. These results systematically replicate basic animal research, and extend the number of responses related to eating in humans that habituate to repeated food presentations. PMID- 9344430 TI - Feather pecking in domestic chicks: its relation to dustbathing and foraging AB - Feather pecking is a serious problem in poultry housing, as it may lead to feather damage, injuries and even mortality. We tested predictions of the two prevalent hypotheses claiming that feather pecking is related to dustbathing and foraging, respectively. Forty-two groups of 30 laying hen chicks, Gallus gallus domesticuswere reared in pens with a slatted floor. Access to sand as a dustbathing substrate and straw as a foraging substrate was varied between groups. The rate of feather pecking was measured in early development up to week 7. The provision of a sand area did not prevent the chicks from developing high rates of feather pecking that caused injuries. Chicks that had access to sand from day 10 showed higher rates of feather pecking than chicks that had access to sand from day 1. The provision of straw to chicks that had developed high rates of feather pecking led to a decrease in this behaviour. Chicks that could use both sand and straw from day 1 on did not show high rates of feather pecking, and no injuries were observed in these groups. There was no significant difference in dustbathing activity between housing conditions characterized by high or low rates of feather pecking. On the other hand, foraging activity was inversely related to the rate of feather pecking, and the occurrence of feather pecking could be delayed from week 4 to week 7 by postponing procedures that led to changes in foraging behaviour. In conclusion, the results show that the presence of an appropriate substrate for dustbathing does not prevent domestic chicks from developing feather pecking. On the other hand, housing conditions that promote foraging behaviour are effective in reducing and preventing feather pecking.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344431 TI - Evolution of sex differences in microhabitat choice and colour polymorphism in Idotea baltica AB - We studied microhabitat choice of colour morphs, causes of sex differences in microhabitat use and colour polymorphism in Idotea balticaa marine isopod living mainly on the brown alga Fucus vesiculosusThe colour morphs differ in frequencies between the sexes and appear to be cryptic on the visually heterogeneous FucusIn this study, no colour-morph-dependent preference for visually matching microhabitats was found. However, in all three experiments conducted, females were found more often on the lower parts of the Fucus than males. The microhabitat choice of the sexes was directed by some character of Fucus itself, not by preferred height within the plant. However, the sexes did not choose differently between upper and lower parts of Fucus as food. The food choice and substrate choice correlated in males but not in females, implying that microhabitat and feeding preferences are more tightly associated in males. We propose that the stronger preference for the less exposed lower parts of Fucus as microhabitat and the lack of correlation between microhabitat and substrate choice in females can be explained in terms of a greater investment in anti predator protection in females than in males. Thus, the sexual difference in microhabitat choice would ultimately result from different strategies maximizing reproductive success in males and females. We suggest that the sexual differences in coloration and colour morph frequency in I. baltica are explained as an adaptation to sex differences in patterns of habitat use.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344432 TI - Self-organizing nest construction in ants: individual worker behaviour and the nest's dynamics AB - We examine nest construction in the ant Leptothorax tuberointerruptus at two levels: (1) the building behaviour of individual workers and (2) the collective properties (temporal and spatial) of the structures they create. We also explore, for the first time explicitly, the linkage between these two levels. Leptothorax tuberointerruptus nests occur in flat cavities which provide the roof and the floor of their dwelling places. Hence, they construct only a peripheral encircling wall, breached by one or more entrance passageways. The wall is constructed brick by brick. This facilitates experimental estimation of the probabilities of individual workers picking up and depositing building material in response to different stimuli. We incorporate both the qualitative and quantitative behavioural rules that works employ during building into a mathematical model. This model confirms that a surprisingly small and simple set of behavioural rules are not only sufficient for wall construction but also for the formation of one or more nest entrances. In addition, this model predicts that the nests of these ants are likely to exhibit interesting dynamics, in which, for example, the tendency to build a new larger nest may lag behind growth of the population that the nest has to house. We present experimental evidence that suggests that this prediction is valid.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344433 TI - Aggression among female lapwings, Vanellus vanellus AB - Social monogamy is the most common pair bond in birds and one hypothesis for its prevalence is that already mated females ('residents') prevent other females from establishing a pair bond with their mates ('competition for male parental care' hypothesis). To investigate this hypothesis we experimentally induced aggressive behaviour in resident female lapwings by presenting a female dummy conspecific, and a male dummy as control, near their nests. Females attacked both dummies. However, the female dummy was attacked more often than the male during the 5-min trials. Attacks on the female dummy were mostly on the ground (88%, N=27 resident females) whereas the male dummy was attacked either by aerial dives (53%) or on the ground (47%, N=24 resident females). Frequency of attacks on the female dummy decreased over the incubation period, whereas there was no such trend with the male dummy. These results suggest that female lapwings attempt to prevent their mates from attracting a new mate and thus try to monopolize their parental care. Other competing hypotheses for the explanation of aggressive behaviour in female lapwings are also discussed but were not supported by our data.1997The Association for the Study of Animal Behaviour PMID- 9344434 TI - Parental state and offspring recognition in the biparental cichlid fish Pelvicachromis pulcher AB - Alloparental care was investigated in the biparental West African cichlid, Pelvicachromis pulcherNon-breeding adults typically consumed young conspecifics but this trait was inhibited in both sexes during reproductive attempts. Alien conspecific young were accepted into the brood if they were of a similar age/developmental stage to the parents' own young but not if they were much older or much younger. If not accepted they were consumed by whichever adult located them. Parents separated from their brood for up to 4 days accepted their young on reunion but separation for more than 4 days resulted in the young being consumed. This latter response occurred if chemical stimuli from the young were available during the separation but not if visual stimuli were available. In this latter case parental responsiveness was maintained. Both sexes of this externally fertilizing species appeared to have the same information about their young and showed the same changes in responsiveness and the same discriminatory abilities.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344435 TI - Evidence for dominant wild female chimpanzees investing more in sons AB - Parents are expected to invest more resources in the offspring gender that promises more grandchildren. In a variety of vertebrate species skewed sex ratio at birth and differential parental investment in sons and daughters have been documented. Wild chimpanzees, Pan troglodytesliving in the Tai National Park, Cote d'Ivoire, were followed for 15 years. This community followed the typical species pattern in that males showed natal philopatry and the sex ratio at birth was almost 1:1. An analysis of 33 inter-birth intervals revealed that dominant females invested about 2 years more in sons, whereas subdominant females invested about 11 months more in daughters. The first difference is significant. Sons of dominant females had higher survival than other youngsters. The benefit of such a facultative investment is discussed. The absence of such a differential investment by mothers in other chimpanzee populations is compatible with an explanation based on variations within the female-biased dispersal pattern in this species and the possible role of maternal condition.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344436 TI - Push or pull: an experimental study on imitation in marmosets AB - A laboratory experiment was conducted in order to explore the possibility of imitation, that is, response learning by observation, in marmosets, Callithrix jacchusInexperienced individuals were allowed to observe a skilful model that demonstrated one of two possible techniques (pushing or pulling a pendulum-door) to get food from inside a wooden box. Their initial manipulative actions, performed when exposed to the box in a subsequent test, were compared with those of naive control subjects (non-observers). The observers showed less exploratory behaviour than the non-observers and, more importantly, some showed a strong tendency to use the demonstrated opening technique in the initial test phase. This initial preference disappeared in the course of five test sessions and the observers converged towards the simpler, alternative solution that was generally preferred by the non-observers. Despite fundamental individual differences in the observer group and the failure to find a significant group effect, the results indicate that marmosets are capable of learning simple motor skills through conspecific observation.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344437 TI - Temporal selectivity of evoked vocal responses of Batrachyla antartandica (Amphibia: Leptodactylidae) AB - The advertisement call of the leptodactylid frog Batrachyla antartandica from southern Chile consists of a train of brief percussive tone pulses whose energy is centred at about 2 kHz. To gain an understanding of the temporal features that are essential for call recognition, playback experiments were conducted with 11 males. Subjects were presented with a synthetic imitation of this signal and variants for which different temporal call parameters were modified systematically. The number of pulses, pulse rate and latency of evoked vocal responses (EVRs) to stimuli having high pulse repetition rates (i.e. 8 and 16 pulses/s) were significantly weaker relative to responses to stimuli having an equal number of pulses but lower pulse rates. A similar, non-significant tendency was observed for a series of stimuli with different pulse rates for which the total stimulus duration was held constant. EVRs also decreased significantly for stimuli having long pulse durations (i.e. 48 and 96 ms) relative to stimuli comprising shorter pulses. No significant differences were observed between EVRs to stimuli for which pulse rise and fall times were varied from 1-20 ms. Responses to calls comprising trains of 10 pulses were weaker compared with stimuli having fewer pulses per train. The selective EVRs of B. antartandica for different temporal parameters contributes to an understanding of the mechanisms involved in call recognition and stress the relevance of temporal processing of sound by males for the emergence of specific patterns of vocal behaviour in anurans.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344438 TI - Worker reproduction and social hierarchies in Leptothorax ants AB - Kin selection theory predicts the existence of potential conflict between queen and workers and among workers concerning the production of males in insect societies with a single, once-mated queen. We investigated the occurrence of reproductive conflict and worker reproduction in both single- and multi-queen colonies of 10 species of the ant genus LeptothoraxIn contrast to previous observations in related species, workers only infrequently engaged in aggressive interactions and did not lay large numbers of eggs in colonies containing queens. Allozyme analyses of queens, workers and males suggest that the contribution of workers to the males produced in colonies with queens is indeed minimal, at least in L. unifasciatusWhen the queens died or were experimentally removed from the colonies, in most species dominance interactions among workers became significantly more frequent and one or several high-ranking workers started to lay eggs. Workers with an increased number of ovarioles per ovary apparently had a reproductive advantage over workers with normal ovaries.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344439 TI - Toad tadpole aggregation behaviour: evidence for a predator avoidance function AB - Two sets of experiments were conducted to determine whether aggregation behaviour, commonly observed in tadpoles of the common toad, Bufo bufohad a predator avoidance function. In the first set of experiments, the distribution of toad tadpoles, of single or mixed sibship, was monitored in large artificial pools, one with fish chemical cues and one without. Aggregation behaviour was determined on the basis of the distribution of the tadpoles under these treatments by using two indices of cohesion: variance/mean ratio and a newly developed swarming index. The two indices were highly correlated. Tadpole groups were more cohesive (1) in the presence of fish chemical cues and (2) with other individuals from a single sibship. In the second set of experiments, tadpoles in different densities and distributions were presented to fish predators in a floating arena which allowed strike rate to be monitored but prevented the fish from capturing any tadpoles. Total strike rate per group increased with increasing group size, but strike rate per individual decreased, implying individuals gain from being in a larger group through dilution, but the group as a whole loses. These results are discussed in terms of selfish herd or cooperative group theories on the behaviour of aggregations.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344440 TI - Do Tengmalm's owls see vole scent marks visible in ultraviolet light? AB - Scent markings (urine and faeces) of small mammals are visible in ultraviolet (UV) light. Diurnal kestrels, Falco tinnunculususe them as a cue to find areas of food abundance. We studied whether vole-eating, nocturnal Tengmalm's owls, Aegolius funereuscan see vole scent marks using UV-vision. In a laboratory experiment, 14 young (less than 6 months old) and 14 adult (more than 6 months old) owls were individually given a choice between four adjacent arenas: (1) an arena with vole urine and faeces in UV light; (2) an arena with vole urine and faeces in visible light; (3) a clean arena in UV light; and (4) a clean arena in visible light. Owls did not prefer any of the four arenas. Our results suggest that Tengmalm's owls probably do not use UV light as a cue to detect vole scent marks.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344441 TI - Field experiments on duration and precision of grey and red squirrel spatial memory AB - Field experiments on squirrel spatial memory under naturalistic conditions showed that grey squirrels, Sciurus carolinensisare capable of relocating precise spatial goals using short range visual cues. These goals were in the form of buried food, similar to natural cache sites, and were relocated to within 5 cm in 62.5% of all attempts after a retention interval of 20 days. Some recall could be demonstrated after more protracted retention intervals (43 and 62 days). These results suggest that grey squirrels' spatial memory is accurate enough to support a successful retrieval strategy. Application of one of the same methods to a population of red squirrels, Sciurus vulgariswhich is not as reliant on stored food, suggested that their spatial memory is less long lasting as predicted by the adaptive specialization hypothesis of cognitive function.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344442 TI - Badge size in collared flycatchers predicts outcome of male competition over territories AB - The evolution of conspicuous coloration is often hypothesized to be driven by sexual selection, where colour traits may function as honest signals of individual abilities in male contest competition and female choice. However, game theory models suggest that colourful badges (i.e. energetically cheap signals) may have no function in sexually selected contests, because the value of the contested resource is too high relative to the costs of fighting. We investigated this assertion by experimentally staging male contests over nest sites (a crucial resource for attracting females) in old (>/=2 years) male collared flycatchers, Ficedula albicollisMales with a relatively large white forehead patch (i.e. a condition-dependent plumage trait displayed in male contests) enjoyed a competitive advantage in disputes over experimentally vacated territories. No other measured morphological variable predicted the outcome of such a dispute. Furthermore, the winners of the disputes acquired a female more quickly than did the losers. Thus, our results suggest that the white forehead patch of male collared flycatchers may function as a badge of status that is also used in sexually selected contests over resources. We suggest that this is because the value of the contested territory may be relatively low compared with the cost of fighting when alternative vacant sites exist in the neighbourhood.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344443 TI - Effect of strange male odour on parental care in lactating female mice AB - This paper analyses the behaviour of lactating female outbred mice, Mus musculus domesticusin the presence of male conspecific odours. When olfactory cues were left in the environment by a sexually naive adult male, a potentially infanticidal animal, the mother took longer to reach her litter following 30 min of separation. Odours left by the sexual partner, by an unknown male of parental status, or by a young naive male did not modify the mother's behaviour, compared with the control situation (absence of male odour). The number of ultrasonic calls of pups varied according to the characteristics of the male but did not modify the behaviour of the dam. Females took longer to reach pups on day 8 of lactation than on days 4 or 12. We suggest adaptive reasons why females take longer to reach pups when the situation is more risky. We tested the hypothesis that the loss of an 8-day-old litter is more expensive, in term of the mother's future reproductive success, than the loss of younger and older litters. Females conceived a new litter within a few days (the inter-birth interval varied according to the age of the litter previously removed) but, even though no difference in size and weight of litters was recorded, females that had the litter removed on day 8 postpartum (compared with days 4 and 12) suffered from a higher mortality rate in the next litter. We suggest that the time the mother takes to reach her pups in the presence of a potentially infanticidal male could represent a measure of parental investment.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344444 TI - Foraging currencies for non-energetic resources: pollen collection by bumblebees AB - Animals sometimes forage for resources whose benefits are not energy-based. In such cases, the benefits and costs of foraging cannot be directly compared, making it difficult to evaluate the currency of fitness maximized by the animal's behaviour. For bees, pollen represents such a resource, because they collect pollen as the sole protein source for developing larvae, rather than as a source of energy. This study compared three currencies that could be maximized during pollen collection by bumblebees (Bombus spp.) foraging on different lupin species (Lupinus spp.): pollen collected per inflorescence, pollen collection rate (gross benefits/time) and pollen collection efficiency (gross benefits/costs). Bees visiting lupin inflorescences begin foraging low on an inflorescence and then move upward through a gradient of increasing pollen availability. To maximize each currency, we predicted that a bee would begin foraging at a different position along a lupin inflorescence, depending on the relative influences of the time and energy costs associated with handling flowers and flight. Based on comparisons of observed and predicted starting positions for seven situations, maximization of gross pollen-collection efficiency predicted observed behaviour better than either pollen collected per inflorescence or gross rate of pollen collection. Such maximization of pollen-collection efficiency would enhance a bee's lifetime input of pollen to her colony, as has been demonstrated for nectar collecting bees. Hence, the economics of foraging for non-energetic resources do not differ qualitatively from foraging for energy.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344445 TI - Spatial patterns of shiny cowbird brood parasitism on chestnut-capped blackbirds AB - Shiny cowbirds, Molothrus bonariensisparasitized a high frequency (average 48%) of nests in five colonies of chestnut-capped blackbirds, Agelaius ruficapillusin Argentina. Two distinct egg morphs occurred, spotted and immaculate white eggs, as well as a few very lightly spotted intermediate eggs. The clear differences between morphs, combined with considerable variation in spotting pattern between spotted morph eggs, made it possible to visually match eggs into groups that were probably laid by single females. Eggs attributed to a single female were more similar in size and shape than eggs attributed to different females. Using egg dimensions and colour patterns to infer spatial patterns of laying by individual females indicated that: (1) individual brood parasites often laid several eggs in the same colony, (2) females also laid eggs in more than one colony, and (3) several females laid eggs in each colony, and often in the same host nests, ruling out the notion that parasites defend exclusive territories with respect to host nests. Multiple cowbird eggs per host nest invariably resulted from several females laying in the same host nest: egg morphs and visual comparisons of spotting patterns at 14 nests indicated that individual females never laid more than a single egg in the same host nest. Experimental parasitism of nests with spotted and white morph eggs, combined with observations of naturally parasitized nests, demonstrated that chestnut-capped blackbirds accept all morphs of cowbird eggs. Since other important host species reject white eggs, however, the adaptive maintenance of the white morph is difficult to explain.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344446 TI - Social interaction alters attraction to competitor's odour in the mouse Mus spretus Lataste AB - When animals defend territories that are large and structurally complex, scent marks alone are unlikely to be reliable signals of a resident's dominance and competitors should require initial proof through direct interaction. This was tested using freshly captured Mus spretus which occupy large non-overlapping ranges in grassland but are strongly attracted to substrate odours from unfamiliar competitors. Choice tests measured time spent investigating and chewing to gain access to paired nestboxes when the entrances were blocked with mesh. Experiment 1 established that mice of both sexes were more strongly attracted to their own odour than to a clean site. Experiment 2 examined choice between the subject's own odour and that of an unfamiliar same-sex competitor both before and after meeting the competitor in a neutral (clean) arena. Prior to interaction, males exerted much effort to gain access to both their own and their unfamiliar competitor's odour. Once relative dominance had been established through agonistic interaction, subordinates avoided their dominant competitor's odour in favour of their own while dominants continued to be attracted to both. There was little aggressive competition between unfamiliar females and relative status did not affect their attraction to a competitor's odour. Females tended to be more attracted to a competitor's odour than to their own prior to interaction but showed less attraction to a competitor's odour post-interaction. A third experiment showed that the odour of an unfamiliar male was more attractive than that from an unfamiliar female, especially to males. The consequences of these responses for maintaining spatial dispersion in this species are discussed.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344447 TI - Stronger territorial responses to frequency modulated coos in collared doves AB - Playback experiments were used to investigate the perception of frequency variations in perch coos by collared doves, Streptopelia decaoctoTerritorial males responded more strongly to modulated than to unmodulated coos. This effect was seen whether all three elements of the coo, or just the first element, were modulated. Modulated coos differed from unmodulated coos in two ways, first by an increased average frequency, and second, by the presence of a discrete change in frequency. We show that it was the change in frequency that was responsible for the level of response to modulated coos. The stronger responses to modulated coos are interpreted as a result of the receiver rating the sender as a stronger competitor.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344448 TI - Patch departure decisions by spice finches foraging singly or in groups AB - The marginal value theorem predicts that when resources are clumped in space, a forager can maximize its rate of intake by deciding to leave a patch when its current feeding rate falls below the average for the habitat. A group version of the model predicts that when rate-maximizing group members share a patch, they should leave sooner, and each with less gain, than single animals exploiting the same patch. We tested these predictions in the laboratory by measuring patch departure decisions of spice finches, Lonchura punctulataexploiting food patches alone or in groups of three under two habitats that require different travel times. As predicted, group members left the patch sooner and with fewer seeds than single foragers. Unlike the model's assumptions, however, birds did not share the patch equally, and their exploitation curves could not be simply derived from those of single foragers. Grouping decreased the effect of travel time on patch exploitation. Moreover, within each group the bird expected to leave first delayed its departure although it collected fewer seeds than the others. This delayed departure could aim to maintain group membership. We noted an increased variability in seed number collected by group members compared with single foragers, which could be a cost of group foraging.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344449 TI - Definitive evidence for cuticular pheromones in a cricket AB - The Orthoptera include many species established as important model systems in the study of animal behaviour, particularly in relation to communication and mating systems. Although most interest has focused on auditory communication, increasing circumstantial evidence suggests that there may be a widespread additional communication channel in the form of cuticular contact pheromones. Using the field cricket, Gryllus bimaculatuswe conducted a behavioural assay which demonstrated that males can distinguish the sex of conspecifics using such a channel. Male response to females (courtship song) was completely abolished by using an organic solvent to remove cuticular hydrocarbons and associated compounds from a stimulus female. It was subsequently restored by painting the washed female with the dissolved extract. This technique controls for the possibility, inherent in previous tests, that the lack of response to washed body parts might be due to the washing process itself. The composition of the cuticles of males and females was analysed using gas chromatography. This revealed that the two sexes differ markedly in the quantities of the majority of the compounds found in the cuticular extract that had previously been shown to be used in mate recognition. This suggests that mate recognition is likely to be due to the relative concentrations of several cuticular compounds, rather than a single 'sex pheromone'. It supports previous assertions of the existence of contact pheromones in the Orthoptera, suggesting that they may be widespread in this group.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344450 TI - Testing chemical defence based on pyrrolizidine alkaloids AB - Pyrrolizidine alkaloids are considered the primary defence mechanism in aposematic ithomiine butterflies and arctiid moths. Despite evidence that pyrrolizidine alkaloids are effective against some invertebrate predators, proof for a protective function of pyrrolizidine alkaloids against vertebrate predators is fragmented. The present work shows that the pyrrolizidine alkaloid monocrot aline is unpalatable to the pileated finch, Coryphospingus pileatusand that the unpalatability is learned through association with a specific colour pattern (blue stripes). In a series of trials, using mealworms as model prey, birds rejected those to which pyrrolizidine alkaloid solution had been applied topically but accepted prey devoid of the alkaloid. Subsequent offerings of prey with pyrrolizidine alkaloid and a painted blue-striped pattern led to consistent rejections by the experimental birds. Birds were then offered blue-striped painted larvae without pyrrolizidine alkaloids ('mimics'), which were rejected at levels similar to the previous trial. The predators learned to recognize the prey as unpalatable items based on their experience in the previous encounters. These results provide evidence for the protective capacity of the pyrrolizidine alkaloid against a vertebrate predator and supports the role of these chemicals in aposematism in the Lepidoptera.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344451 TI - The predatory response of a stalking spider, Plexippus paykulli, to camouflage and prey type AB - When approaching prey, a stalking predator should consider trade-offs between the probabilities of early detection (by the prey, before the strike), spontaneous departure (of prey, before the strike), prey escape (following the strike) and interference (by rivals or predators). In this study we tested the response of a jumping spider, Plexippus paykullito a background with two different camouflaging properties, and two different prey types (maggots versus adult house flies). Spiders jumped towards adult house flies from greater distances on a non camouflaging background, but background colour had no effect on jumping distance when the prey were maggots. Spiders stalking both prey types approached more slowly when camouflaged. Our experiments suggest that jumping spiders may be responding to changes in the trade-off relationships between the probabilities of early detection, spontaneous departure, escape and interference.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344452 TI - Division of labour in a lower termite: the majority of tasks are performed by older workers AB - Division of labour among workers was investigated in the lower termite, Reticulitermes fukienensisWorkers were separated into three age groups based on size, small workers being the youngest, medium workers intermediate and large workers the oldest. Workers were then compared in behavioural assays for the degree to which they would carry out specific tasks, which included: (1) foraging related tasks; (2) care of eggs, larvae and the queen; and (3) some other important behaviours including burying corpses, alarm-giving and time spent stationary. All tasks were performed by two or all three of the size-groups of workers. Hence evidence does not support the hypothesis of tasks being discretely allocated among different instars in termites and this having evolved towards the extreme of one caste for every task. The oldest workers (i.e. large workers) carried out the highest frequencies of all tasks investigated. This contrasts with the social Hymenoptera, where younger workers specialize in some tasks (especially brood and queen care). The results suggest a new pattern for social insects for division of labour among workers.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344453 TI - Sexually selected vigilance behaviour of the grey partridge is affected by plasma androgen levels AB - In the grey partridge, Perdix perdixvigilance and calling activity are two sex dimorphic behaviours that are critical for mate choice. To ascertain whether circulating levels of testosterone directly affect vigilance (i.e. the occurrence of upright alert posture), we compared vigilance scores of testosterone-implanted versus control males both during the normal activity of the flock and after the passage of a raptor silhouette; in the latter case, the calling activity was also recorded. Hormone-treated males were more vigilant than controls in both experimental situations. Vigilance was correlated with calling rate. Testosterone seems to act as a link, relating conspicuous behaviours involved in sexual selection to male quality and physical condition, because of the costs of having high levels of both signalling and androgens.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344454 TI - Distance and the presentation of visual stimuli to birds AB - Artificial visual stimuli in the form of photographs, video sequences and computer-generated images are increasingly being used to explore the visual world of birds but their use is controversial as it is still not clear whether birds see them in the same way that humans do. While differences between bird and human colour vision may be one problem with using such artificial images, another and potentially even more important difficulty is the distance at which stimuli are presented. An experiment is described in which hens, Gallus gallus domesticuswere trained to move towards one of two real objects viewed at two different distances. Even for real objects, discrimination levels were better when the hens were allowed to view the stimuli from 5-25 cm than when they were forced to choose at 120 cm and this correlated with their ability to transfer to photographs of the same objects at different distances. In a colour discrimination at a short distance, five out of seven hens showed 100% correct responses when first shown photographs of real objects that they had previously learnt to discriminate. The results suggest that photographs can be used as substitutes for real stimuli but that care should be taken over the distance at which they are presented. The results are discussed in relation to the visual behaviour of birds and differences in functioning of their frontal and lateral visual fields.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344455 TI - Using randomization techniques to analyse fluctuating asymmetry data AB - No Abstract Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344456 TI - Fluctuating asymmetry, mate choice and experimental designs AB - No Abstract Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344457 TI - Experimental design and the signalling properties of fluctuating asymmetry AB - No Abstract Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour PMID- 9344459 TI - Molecular cloning, sequencing, and expression in Escherichia coli of mouse flavin containing monooxygenase 3 (FMO3): comparison with the human isoform. AB - The sequence of mouse flavin-containing monooxygenase 3 (FMO3) was obtained from several clones isolated from a mouse liver cDNA library. The nucleotide sequence of mouse FMO3 was 2020 bases in length containing 37 bases in the 5' flanking region, 1602 in the coding region, and 381 in the 3' flanking region. The derived protein sequence consisted of 534 amino acids including the putative flavin adenine dinucleotide and NADP+ pyrophosphate binding sites (characteristic of mammalian FMOs) starting at positions 9 and 191, respectively. The mouse FMO3 protein sequence was 79 and 82% identical to the human and rabbit FMO3 sequences, respectively. Mouse FMO3 was expressed in Escherichia coli and compared to E. coli expressed human FMO3. The FMO3 proteins migrated with the same mobility ( approximately 58 kDa) as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The expressed FMO3 enzymes (mouse and human forms) were sensitive to heat and reacted in a similar manner toward metal ions and detergent. Catalytic activities of mouse and human FMO3 were high toward the substrate methimazole; however, in the presence of trimethylamine and thioacetamide, FMO-dependent methimazole oxidation by both enzymes was reduced by greater than 85%. Other substrates which inhibited methimazole oxidation were thiourea and thiobenzamide and to a lesser degree N,N-dimethylaniline. When probed with mouse FMO3 cDNA, FMO3 transcripts were detected in hepatic mRNA samples from female mice, but not in samples from males. FMO3 was detected in mRNA samples from male and female mouse lung, but FMO3 message was not detected in mouse kidney sample from either gender. Results of immunoblotting confirmed the tissue- and gender-dependent expression of mouse FMO3. PMID- 9344458 TI - A structural requirement of zinc for the folding of recombinant link protein. AB - We have cloned and ligated a full-length bovine link protein (LP) in the pMAL-c2 vector and overexpressed it in fusion with maltose-binding protein (MBP) in Escherichia coli. We have demonstrated dose-dependent binding of MBP/LP to biotinylated hyaluronan in a dot blot assay. A greater percentage of the expressed fusion protein was soluble, monomeric, and undegraded when the growth temperature was lowered, the growth medium was supplemented with zinc, and metal chelators were omitted from the lysis buffers. Similar effects were observed when we tested the effects of lower growth temperature and zinc supplementation on another construct consisting of MBP in fusion with the first proteoglycan tandem repeat of LP. Our results suggest zinc may be necessary for the folding and disulfide bond formation of recombinant LP. In addition, a greater amount of monomeric MBP/LP produced at 27 degrees C with zinc supplementation bound to biotinylated hyaluronic acid-binding region of aggrecan than MBP/LP produced at 27 or 37 degrees C without zinc. This suggests that recombinant LP may have a conformational requirement for zinc necessary for binding to aggrecan. Factor Xa cleavage of MBP/LP expressed in the presence of zinc yielded much more intact LP product than cleavage of MBP/LP expressed without zinc. These data indicate a structural role of zinc that allows MBP/LP to fold in a manner such that it is resistant to proteolytic degradation. PMID- 9344460 TI - Differential effects of heparin and glucose on structural conformation of human alpha1 antitrypsin: evidence for a heparin-induced cleaved form of the inhibitor. AB - alpha1 Antitrypsin (alpha1AT) is the archetypal member of the serpin superfamily. Current knowledge of its inhibitory mechanism does not provide for any heparin induced enhancement of serine proteinase inhibition. Since previous results have shown that an apparently altered alpha1AT form may be purified from the plasma of insulin-dependent diabetics by means of heparin-affinity chromatography, in the present work the possibility was tested that heparin at various concentrations modifies the structural conformation and function of human alpha1AT in the absence and presence of glucose, used at concentrations of 15 mM to mimick mild hyperglycemic conditions. Heparin was observed to bind strongly to alpha1AT, causing maximal enhancement of tryptophan fluorescence emission at 50 microg/ml, mostly in the presence of glucose. Circular dichroism spectra revealed that heparin with glucose caused the most relaxed, ordered structure of the inhibitor with increased heat stability. Modification in conformation was accompanied by loss of inhibitory activity, as demonstrated by the inability of alpha1AT to block bovine trypsin in the specific assay and by alterations of its immunological properties. However, despite inactivation, in the presence of heparin-both with and without glucose-alpha1AT was still able to bind trypsin, as revealed by inhibitor-to-proteinase complexes visible in both SDS- and nondenaturing electrophoreses. These complexes showed the same feature regardless of trypsin concentration and differed from those formed at a molar excess of the inhibitor in the absence of heparin, since they underwent rapid, intense fragmentation accompanied by complete loss of the secondary structure of the inhibitor. Even in the absence of trypsin, cleavage of alpha1AT was also observed to occur at both Val321- and Glu344- in the primary sequence of the inhibitor in the presence of 50 microg/ml heparin, with and without glucose. These results suggest that heparin binding to alpha1AT causes profound structural changes in the molecule, involving both the expulsion of the reactive site out of the molecule plane and a relaxed, heat-stable form of the inhibitor, rendered a substrate for the proteinase. Although glucose apparently does not affect alpha1AT functioning, it does enhance the effects of heparin on the alpha1AT structure. The possibility is discussed that, while heparin and glucose binding occurs at different sites on alpha1AT, glucose favors heparin binding by inducing a partially relaxed form in the inhibitor. Differences in structure and charge between the two substances account for both different individual effects on alpha1AT and the predominance of the effects of heparin. PMID- 9344461 TI - Naturally occurring variants of human milk bile salt-stimulated lipase. AB - Analysis of milk samples from a number of lactating women revealed molecular variants of bile salt-stimulated lipase (BSSL) of both lower and higher molecular mass than that commonly occurring. In contrast to previous observations, we report on individuals having only a variant of lower mass, both one of lower and one of common mass, or both one of lower and one of higher mass of the lipase. From two individuals we purified the lower molecular mass BSSL variant and characterized it. The amount of lipase in the milk of these two individuals was considerably less than average (mean of 10 women with BSSL of the most common molecular mass). The BSSL variant of lower mass showed the same bile salt activation, pH dependency, temperature stability as those most commonly occurring. We could localize the difference in mass to the large O-glycosylated repeat sequence close to the C-terminus of the protein. With respect to all characteristics studied, the BSSL variant of higher mass was also similar to that most commonly ocurring. Again, the difference in mass could be localized to the repeat region of the protein. Hence, it appears as if the repeat region, normally carrying 16 repeats of 11 amino acids each, varies in size between individuals. PMID- 9344462 TI - Expression of protein kinase C-beta promotes the stimulatory effect of phorbol ester on phosphatidylethanolamine synthesis. AB - Stimulation of phosphatidylethanolamine (PtdEtn) synthesis by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) has reportedly been found only in hepatocytes expressing the alpha-, betaII-, epsilon-, and zeta-PKC isozymes. In contrast, stimulation of phosphatidylcholine synthesis by PKC activators, known to be mediated by PKC-alpha, is widespread in mammalian cells. In this work, various cell lines exhibiting characteristic differences in their PKC systems were used to determine the role of specific PKC isozymes in the mediation of PMA effect on PtdEtn synthesis. In NIH 3T3 fibroblasts, which express high levels of PKC-alpha but none of the beta (betaI or betaII) isoforms, PMA did not stimulate PtEtn synthesis. In contrast, in Rat-6 fibroblasts overexpressing PKC-betaI, 10-100 nM PMA considerably (1.7- to 2.6-fold) enhanced PtdEtn synthesis. In wild-type or multidrug resistant MCF-7 human breast carcinoma cells, which express PKC-alpha and PKC-betaII (to varying extents) but not PKC-betaI, PMA had only small or no effects on PtdEtn synthesis. In contrast, in MCF-7 cells overexpressing PKC-alpha, and as a consequence also expressing the betaI- and betaII-PKC isoforms, PMA effectively stimulated the synthesis of PtdEtn. Finally, in HL60 human leukemia cells, which contains PKC-betaII as the major PKC isoform, PMA again stimulated PtdEtn synthesis. The results establish that while stimulation of PtdEtn synthesis by PMA occurs only in selected cell lines, this phenomenon is not restricted to hepatocytes. Furthermore, the data indicate that expression of either PKC-betaI or PKC-betaII, but not PKC-alpha, correlates with the effect of PMA on PtdEtn synthesis. Overall, these observations strongly suggest that regulation of PtdEtn and PtdCho synthesis by PMA involves separate PKC isozymes, i.e., PKC-beta and PKC-alpha, respectively. PMID- 9344463 TI - ESR identification of free radicals formed from the oxidation of catechol estrogens by Cu2+. AB - Catechol estrogens are genotoxic, indirectly through redox cycling mechanisms leading to oxidative DNA damage and directly by formation of quinone-DNA adducts. Previously, we demonstrated that Cu2+ can oxidize estradiol (E2) catechols, establishing a copper redox cycle leading to the formation of DNA strand breaks. The goal of this study was to use electron spin resonance techniques to identify the free radical intermediates formed. The 2- and 4-OH catechols of E2 and ethinyl estradiol (EE) were oxidized to semiquinone intermediates, stabilized by Mg2+, when incubated with Cu2+. The 4-OH-EE semiquinone decayed more slowly than the 2-OH-EE semiquinone. Using the spin trap alpha-(4-pyridyl-1-oxide)-N-tert butylnitrone, 4-OH-E2 plus Cu2+ generated hydroxyl radicals at a greater rate than 2-OH-E2 plus Cu2+. Formation of hydroxyl and methyl radical adducts was detected, using 5,5-dimethyl-1-pyrroline-N-oxide as the spin trap, when 2-OH-E2 was incubated with Cu2+ and 1% dimethyl sulfoxide. This was inhibited by the Cu1+ chelator bathocuproinedisulfonic acid and catalase. These data demonstrate that the oxidation of estrogen catechols by Cu2+ leads to a Cu-dependent mechanism of hydroxyl radical production via a hydrogen peroxide intermediate and suggest a mechanism for estrogen-associated site-specific DNA damage and mutagenesis. PMID- 9344464 TI - Functional studies of yeast-expressed human heart muscle carnitine palmitoyltransferase I. AB - Long-chain fatty acids are the primary source of energy production in the heart. Carnitine palmitoyltransferase I (CPT-I) catalyzes the first reaction in the transport of long-chain fatty acids from the cytoplasm to the mitochondrion, a rate-limiting step in beta-oxidation. In this study, we report the functional expression of the human heart/skeletal muscle isoform of CPT-I (M-CPT-I) in the yeast Pichia pastoris. Screening of a human heart cDNA library with cDNA fragments encoding the rat heart M-CPT-I resulted in the isolation of a single full-length human heart M-CPT-I cDNA clone. The clone has an open reading frame of 2316 bp with a 5' untranslated region of 38 bp and a 256-bp 3' untranslated region with the poly(A)+ addition sequence AATAAA. The predicted protein has 772 amino acids and a molecular mass of 88 kDa. Northern blot analysis of mRNAs from different human tissues using the human M-CPT-I cDNA as a probe revealed an abundant transcript of approximately 3.1 kb that was only present in human heart and skeletal muscle tissue. Expression of the human M-CPT-I cDNA in P. pastoris, a yeast with no endogenous CPT activity, produced an 80-kDa protein that was located in the mitochondria. Isolated mitochondria from the M-CPT-I expression strain exhibited a malonyl-coenzyme A (CoA)-sensitive CPT activity that was detergent labile. The I50 for malonyl-CoA inhibition of the yeast-expressed M-CPT I was 69 nM, and the Kms for carnitine and palmitoyl-CoA were 666 and 42 microM, respectively. The I50 for malonyl-CoA inhibition of the heart enzyme is 30 times lower than that of the yeast-expressed liver CPT-I, and the Km for carnitine is more than 20 times higher than that of the liver CPT-I. This is the first report of the expression of a heart CPT-I in a system devoid of endogenous CPT activity and the functional characterization of a human heart M-CPT-I in the absence of the liver isoform and CPT-II. PMID- 9344466 TI - Rat harderian gland porphobilinogen deaminase: characterization studies and regulatory action of protoporphyrin IX. AB - Properties of purified porphobilinogen deaminase (PBG-D; EC 4.3.1.8) from rat harderian gland are here presented. The enzyme behaves as a monomer of Mr 38 +/- 2 kDa and is optimally active at pH 8.0-8.2. Its activation energy, determined by an Arrhenius plot, is 76.1 kJ/mol. Initial velocity studies showed a linear progress curve for uroporphyringen I formation and a hyperbolic dependence of the initial rate on substrate concentration, indicating the existence of a sequential displacement mechanism. Apparent kinetic constants, Km and Vm, calculated at 37 degrees C and pH 8.0 were 1.1 microM and 170 pmol/min mg, respectively. The pH dependence of the apparent kinetic parameters revealed the ionization of residues with pKAES and pKBES of 7.4 +/- 0.1 and 8.6 +/- 0.1, respectively, and a pKE value of 8.0 +/- 0.1. Incubation of PBG-D with 5.0 mM N-ethylmaleimide and 5.0 mM 5,5'-dithiobis(2-nitrobenzoic acid) at pH 8.0 led to inhibitions of 70 and 50%, respectively. The effect of pH, as well as the effect of thiol reagents, on enzyme activity strongly suggests the involvement of cysteine residue(s) in the mechanism of uroporphyrinogen I biosynthesis, in both the catalytic reaction and the substrate binding. Rat harderian gland PBG-D activity decreased with increasing concentrations of protoporphyrin IX, reaching a 40% inhibition at the in vivo concentration of the porphyrin and 7 microM PBG. Even at saturating concentrations of substrate, inhibition by protoporphyrin was not completely reversed. So, accumulated porphyrin may act as an regulator of PBG-D activity in rat harderian gland. PMID- 9344465 TI - Interaction of matrix metalloproteinases-2 and -9 with pregnancy zone protein and alpha2-macroglobulin. AB - The binding of matrix metalloproteinases-2 and -9 to pregnancy zone protein and alpha2-macroglobulin was studied. The binding was demonstrated by formation of dimeric as well as tetrameric complexes of pregnancy zone protein and by the formation of alpha2-macroglobulin complexes with fast and intermediate mobility in native gel electrophoresis. The complex formation was confirmed by the use of 125I-labeled matrix metalloproteinase-2. The cleavage sites in the "bait" regions following formation of high-molecular-weight complexes of matrix metalloproteinases with the alpha-macroglobulins were determined by protein sequence analysis. Pregnancy zone protein was cleaved at Thr693-Tyr694 and alpha2 macroglobulin at Gly679-Leu680 and Arg696-Leu697 by matrix metalloproteinase-2. Matrix metalloproteinase-9 cleaved alpha2-macroglobulin at the same site as matrix metalloproteinase-2, but cleavage of pregnancy zone protein was at Leu753 Ser754. The sequences of the bands, visualized in the SDS gel, of approximately 90 and 165 kDa or higher molecular weight complexes were the same. This indicates that the matrix metalloproteinases cleaved the inhibitors with or without binding to them. The present results suggest that matrix metalloproteinases-2 and -9 may interact with pregnancy zone protein and alpha2-macroglobulin in vivo. PMID- 9344467 TI - Purification and characterization of UBP6, a new ubiquitin-specific protease in Saccharomyces cerevisiae. AB - Ubiquitin-specific protease-6 (UBP6) in Saccharomyces cerevisiae was expressed in Escherichia coli and purified from the cells using 125I-labeled ubiquitin-alphaNH MHISPPEPESEEEEEHYC as a model substrate. The purified UBP6 behaved as a 58-kDa under both nondenaturing and denaturing conditions, indicating that the enzyme comprises a single polypeptide. It was maximally active at pH levels between 8.5 and 9, but showed little or no activity at pH below 7 and above 9.5. As with other UBPs, its activity was strongly inhibited by sulfhydryl-blocking reagents, such as N-ethylmaleimide, and by ubiquitin-aldehyde. In addition to the model substrate, UBP6 hydrolyzed ubiquitin-alphaNH-protein extensions, such as the ubiquitin-alphaNH-carboxyl extension protein of 80 amino acids and ubiquitin alphaNH-dihydrofolate reductase, but not poly-His-tagged diubiquitin. It was also capable of releasing free ubiquitin from branched polyubiquitin chains that are ligated to proteins through epsilonNH-isopeptide bonds, although to a limited extent. These results suggest that UBP6 may play an important role in the generation of free ubiquitins and certain ribosomal proteins from ubiquitin ribosomal fusion proteins as well as in deubiquitination of certain polyubiquitinated proteins targeted for degradation by the 26S proteasomes. PMID- 9344468 TI - Subunit fusion confers tolerance to peptide insertions in a virus coat protein. AB - An octapeptide sequence called Flag was inserted into the bacteriophage MS2 coat protein at two different locations and its effects on protein folding and virus assembly were determined. Assays of the translational repressor and capsid assembly functions of the recombinants show that when the peptide is inserted at its N-terminus coat protein folds properly into the form that binds RNA (i.e., the dimer), but is defective for capsid assembly. On the other hand, a recombinant protein which is expected to display the Flag insertion as a surface loop does not fold correctly and, as a consequence, is proteolytically degraded. Genetic fusion of the two subunits of the coat dimer results in a protein considerably more tolerant of these structural perturbations and mostly corrects the defects accompanying Flag peptide insertion. Increased resistance of the single-chain coat protein to urea denaturation indicates that the fused dimer is substantially more stable than wild type. Covalent joining of subunits of oligomers probably represents a general strategy for engineering increased protein stability. PMID- 9344470 TI - Multiple levels of regulation of Escherichia coli succinyl-CoA synthetase. AB - Concentrations of GDP, which are expected to bind to the catalytic site and inhibit the autophosphorylation of succinyl-CoA synthetase (SCS) when NTP is used as a substrate, were found to increase the level of phosphoenzyme formed. The ability of GDP to do so is dependent upon the presence of a protein distinct from SCS. The effector protein could be separated from SCS by ammonium sulfate fractionation. Reconstitution experiments show that the protein inhibits SCS, that the inhibition is relieved by GDP, and that the inhibitor recognizes both Escherichia coli and eukaryotic forms of SCS. The inhibitor is itself regulated by the conditions used to grow the bacteria and in a manner that appears distinct from that of SCS. PMID- 9344469 TI - Escherichia coli flavodoxin sepharose as an affinity resin for cytochromes P450 and use to identify a putative cytochrome P450c17/3beta-hydroxysteroid dehydrogenase interaction. AB - Flavodoxin Sepharose (Fld Sepharose), a reagent originally developed to demonstrate an interaction between native Escherichia coli Fld and cytochrome P450c17, has been synthesized, using highly expressed (7 micromol Fld/liter E. coli culture) recombinant E. coli Fld, for use as an affinity resin for microsomal cytochromes P450. As a test of the specificity of Fld Sepharose, we have examined the utility of this resin for purification of P450c17 and P450c21 from a relatively crude mixture of solubilized adrenocortical microsomal proteins. Chromatography of this mixture on Fld Sepharose resulted in a threefold enrichment of cytochrome P450 specific content without spectrally detectable P450 denaturation. Electrophoretic and immunoblot analyses of fractions eluted from the Fld Sepharose column revealed the presence of P450c17 and P450c21, both of which were sufficiently pure, after SDS-PAGE, for identification by N-terminal sequence analysis. Intriguingly, a major protein copurifying with P450c17 and P450c21 was identified as 3beta-hydroxysteroid dehydrogenase (3beta-HSD) which was subsequently found not to directly bind Fld Sepharose. Purified bovine 3beta HSD covalently linked to Sepharose can bind recombinant bovine P450c17, an interaction which is partially disrupted upon mild heat denaturation of P450c17 or by the nonionic detergent Emulgen. This interaction, however, does not appear to affect P450c17 hydroxylase and lyase activities as measured in vitro. From these results, we propose that 3beta-HSD and P450c17 may associate, perhaps as part of a steroidogenic complex, in the endoplasmic reticulum. PMID- 9344471 TI - Hydrophobic ion pairing as a method for enhancing structure and activity of lyophilized subtilisin BPN' suspended in isooctane. AB - The use of enzymes in low water environments permits reactions to occur that are difficult or impossible in aqueous solution. In this manner, proteases can be used to form, rather than hydrolyze, ester and amide linkages. Presumably, the native-like structure of the enzyme must remain intact for catalysis to transpire. However, little is known regarding the integrity of the overall structure of lyophilized proteins suspended in organic media. In this study, the structural changes that occur during the freeze-drying process and those effected by suspension in the organic solvent were examined. Using Fourier-transform infrared spectroscopy, the secondary structure of lyophilized subtilisin BPN' was monitored and correlated to the level of enzymatic activity when suspended in isooctane. In addition, the ability of ionic detergents to stabilize subtilisin BPN' via ion pairing was evaluated. It was found that subtilisin unfolds to some degree during lyophilization, whether it is ion paired or not. Furthermore, there are structural changes observed when the enzyme is placed in isooctane, although the effects are less with ion-paired subtilisin. This higher level of retention of secondary structure results in increased enzymatic activity. PMID- 9344472 TI - In situ properties of Helicobacter pylori aspartate carbamoyltransferase. AB - The kinetic and regulatory properties of aspartate carbamoyltransferase (ACTase) of the human pathogen Helicobacter pylori were studied in situ in cell-free extracts. The presence of enzyme activity was established by identifying the end product as carbamoylaspartate using nuclear magnetic resonance spectroscopy. Activity was measured in all strains studied, including recent clinical isolates. Substrate saturation curves determined employing radioactive tracer analysis or a microtiter colorimetric assay were hyperbolic for both carbamoyl phosphate and aspartate, and there was no evidence for substrate inhibition at higher concentrations of either substrate. The apparent Km were 0.6 and 11.6 mm for carbamoyl phosphate and aspartate, respectively. Optimal pH and temperature were determined as 8.0 and 45 degrees C. Activity was observed with the l- but not the d-isomer of aspartate. Succinate and maleate inhibited enzyme activity competitively with respect to aspartate. The carbamoyl phosphate analogues acetyl phosphate and phosphonoacetic acid inhibited activity in a competitive manner with respect to carbamoyl phosphate. With limiting carbamoyl phosphate purine and pyrimidine nucleotides, tripolyphosphate, pyrophosphate, and orthophosphate inhibited competitively at millimolar concentrations. Ribose and ribose 5 phosphate at 10 mm concentration showed 20 and 35% inhibition of enzyme activity, respectively. N-Phosphonoacetyl-l-aspartate (PALA) was the most potent inhibitor studied, with 50% inhibition of enzyme activity observed at 0.1 microM concentration. Inhibition by PALA was competitive with carbamoyl phosphate (Ki = 0.245 microM) and noncompetitive with aspartate. The kinetic and regulatory data on the activity of the H. pylori enzyme suggest it is a Class A ACTase, but with some interesting characteristics distinct from this class. PMID- 9344473 TI - A common duplication in the lysyl hydroxylase gene of patients with Ehlers Danlos syndrome type VI results in preferential stimulation of lysyl hydroxylase activity and mRNA by hydralazine. AB - Patients with Ehlers Danlos Syndrome type VI (EDS VI) are biochemically characterized by a deficiency of lysyl hydroxylase (LH), an enzyme that hydroxylates lysine residues required in the formation of stable crosslinks in collagen biosynthesis. Recently, in 19% of 35 EDS VI families, a duplication of seven exons in the LH gene has been identified as a common cause of EDS VI. We have observed that in fibroblasts from patients with this duplication mutation, administration of hydralazine, an iron-chelating agent, and ascorbate, a cofactor for LH activity, stimulates LH activity and its mRNA significantly more than in other EDS VI patients who do not have this duplication. Administration of these reagents, either singly or in combination, to fibroblasts from five patients homozygous for the duplication stimulated the low basal level of LH activity (<20% of normal) by five- to sixfold (hydralazine +/- ascorbate) and by twofold (ascorbate alone) at 72 h. This paralleled the increase in the steady-state level of mRNA for LH measured in similarly treated fibroblasts from four of these patients. In contrast, the activity of LH in fibroblasts from six other EDS VI patients and the mRNA from four of these patients who did not have the duplication were increased only three- to fourfold by hydralazine +/- ascorbate. The mechanism for the preferential stimulation of LH activity and mRNA by hydralazine in the EDS VI cells carrying the duplication is unknown, but it could be attributed to the presence of, for example, an enhancer sequence within the duplicated region of the LH gene. PMID- 9344474 TI - Heterologous expression, purification, and properties of diol dehydratase, an adenosylcobalamin-dependent enzyme of Klebsiella oxytoca. AB - Recombinant adenosylcobalamin-dependent diol dehydratase of Klebsiella oxytoca overexpressed in Escherichia coli was purified to homogeneity. The enzyme has a low solubility and was extracted from the crude membrane fraction with 1% Brij 35 in a high recovery. Subsequent chromatography on DEAE-cellulose resulted in 4.9 fold purification of the enzyme in an overall yield of 65%. The enzyme thus obtained showed specific activity comparable to that of the wild-type enzyme of K. oxytoca. The apparent molecular weight determined by nondenaturing gel electrophoresis on a gradient gel was 220,000. The enzyme consists of equimolar amounts of the three subunits with apparent Mr of 60,000 (alpha), 30,000 (beta), and 19,000 (gamma). Therefore, the subunit structure of the enzyme is most likely alpha2beta2gamma2. The recombinant enzyme was also separated into components F and S upon DEAE-cellulose chromatography in the absence of substrate. Components F and S were identified as the beta subunit and alpha2gamma2 complex, respectively. Apparent Km for adenosylcobalamin, 1,2-propanediol, glycerol, and 1,2-ethanediol were 0.83 microM, 0.08 mM, 0.73 mM, and 0.56 mM, respectively. The three genes encoding the subunits of diol dehydratase were overexpressed individually or in various combinations in Escherichia coli. The alpha and gamma subunits mutually required each other for correct folding forming the soluble, active alpha2gamma2 complex (component S). Expression of the beta subunit in a soluble, active form (component F) was promoted by coexpression with both the alpha and gamma subunits, probably by coexistence with component S. These lines of evidence indicate that each subunit mutually affects the folding of the others in this heterooligomer enzyme. PMID- 9344475 TI - Inhibition of oxidation of low-density lipoprotein by a novel antioxidant, BO 653, prepared by theoretical design. AB - 2,3-Dihydro-5-hydroxy-2,2-dipentyl-4,6-di-tert-butyl-benzofuran (BO-653) is a novel antioxidant synthesized by theoretical designing based on the previous experimental findings and consideration. The antioxidant activities of BO-653 against the oxidative modification of low-density lipoprotein (LDL) induced by free radicals were studied. BO-653 was consumed faster than endogenous alpha tocopherol and inhibited the formation of lipid hydroperoxides, which was observed during the consumption of alpha-tocopherol. Doxyl stearic acids incorporated into LDL as spin probes competed with the antioxidants in scavenging radicals. It was found that the efficacy of radical scavenging by alpha tocopherol became smaller as the radical went deeper into the interior of LDL particle, whereas that by BO-653 did not change. Ascorbic acid in the aqueous phase spared alpha-tocopherol efficiently during oxidation. On the other hand, the sparing effect of ascorbic acid for BO-653 was not remarkable, unlike that for alpha-tocopherol, which implied different locations of radicals derived from BO-653 and alpha-tocopherol within the LDL particle. It was concluded that BO-653 protected LDL from oxidative modification efficiently by scavenging peroxyl radicals and by reducing alpha-tocopheroxyl radicals and that this novel antioxidant might act as a potent inhibitor of development of atherosclerosis. PMID- 9344476 TI - Protein expression, characterization, and regulation of CYP4F4 and CYP4F5 cloned from rat brain. AB - We previously reported the cDNA cloning of three new forms of P450, CYP4F4, CYP4F5, and CYP4F6, from a rat brain cDNA library. In the present study, we expressed CYP4F4 and CYP4F5 in Escherichia coli using the pCWOri expression vector with a modification of their N-terminal amino acid sequences and the incorporation of a C-terminal [His]4 tag to aid in purification. CYP4F5 recombinant protein was purified to a specific content of 7.7 nmol/mg protein from the membrane fraction of E. coli and showed omega-hydroxylation activity toward leukotriene B4 (LTB4), a chemical mediator of inflammation. On the other hand, the solubilized membrane fraction of CYP4F4-expressed recombinant protein catalyzed the omega-hydroxylation of prostaglandin A1, prostaglandin E1, and 6 trans-LTB4 as well as LTB4. The effects of the peroxisome proliferator, clofibrate, on mRNA expression of CYP4F4, 4F5, and 4F6 were studied by Northern blot analysis. The expression levels of the mRNA of these CYP4Fs were shown to be reduced by clofibrate in liver. PMID- 9344477 TI - Patterns of sentence comprehension in aphasia: a consideration of three hypotheses. AB - Three hypotheses concerning the functional source of aphasic patients' difficulty comprehending semantically reversible sentences were tested using declarative sentences in active and passive voice and sentences with center-embedded relative clauses. Each of the three hypotheses is predicated on relative patterns of impairment and sparing of patient performance on these (and other) sentence types, yet the three hypotheses make somewhat different predictions about performance patterns across these types. Results from 5 Broca's aphasic patients were not consistent with the predictions of the linguistically motivated Trace Deletion Hypothesis or of a hypothesis based on an impairment involving grammatical morphemes. The hypothesis that aphasic comprehension impairments reflect a general limitation of working memory capacity was given partial support by the ordinal pattern of difficulty for a mixed group of 10 patients, but failed to account for patterns obtained from individual patients. Results are interpreted as having relevance for methodological as well as theoretical aspects of research on aphasic sentence comprehension. PMID- 9344478 TI - Follow-up study of a right- and a left-hemispherectomized child: implications for localization and impairment of language in children. AB - Two hemispherectomized girls, one operated on the right, the other on the left, were followed from time of surgery until 9 and 10 years of age and compared with respect to course of language acquisition following surgery. At conclusion of follow-up, receptive and expressive language, phoneme perception and production, and sentence processing of the two hemispherectomized children were compared with those of two control groups of similar age, one developing language normally, the other language-impaired. The left-hemispherectomized child's abilities were similar to those of the language-impaired children; the right-hemispherectomized child's abilities resembled those of the language-normal children. Implications for localization of developmental anomalies in language-impaired children are discussed. PMID- 9344479 TI - Cerebral hemispheric control of speech during the intracarotid sodium amytal procedure: An acoustic exploration. AB - In this study we investigated lateralized control of speech during the intracarotid amobarbital procedure. Vowel segments were extracted from recordings made during two separate amobarbital procedures and involving two patients. Subjects were right-handed, presented with focal left mesial temporal epileptogenic foci. Age of onset of seizure disorders was 1.5 years for one subject and 16 years for the other. Recorded pre- and postinjection speech samples were digitized. Analyses were conducted on formants 1 and 2 (F1, F2) measures to determine the extent of formant fluctuation in the time course of the IAP. Preliminary results showed, for these two cases, that the left hemisphere is involved in the control of both F1 and F2 and the right in the control of F2 only. The data reveal the potential of coupling the IAP procedure and the acoustical analysis of speech in the study of cerebral control of speech. PMID- 9344480 TI - The relation of planum temporale asymmetry and morphology of the corpus callosum to handedness, gender, and dyslexia: a review of the evidence. AB - Asymmetry of the planum temporale in relation to handedness, gender, and dyslexia is reviewed. The frequency of rightward asymmetry is rather higher than are estimates of the proportion of right hemisphere speech representation in the general population. Conversely, the frequency of leftward asymmetry is lower than the proportion of the population with left hemisphere speech. Neuro-anatomic asymmetry may relate more to handedness than to language lateralization. There are suggestions that neuroanatomic asymmetry is reduced in females compared to males but the data are inconclusive. Reports concerning handedness and gender differences in callosal structure are conflicting but, as with planum asymmetry, any effect of handedness is as likely to relate to degree as to direction of handedness. It has been reported that the plana are more often symmetrical in size or larger on the right side among dyslexics than controls but this has not always been found. However, greater frequency of atypical (a)symmetry of the planum in dyslexia would be consistent with the absence of a factor which, when present, biases the distribution of planum asymmetry toward the left (and handedness towards the right) as hypothesized by Annett (1985). Studies of the size of the corpus callosum in dyslexia have produced conflicting findings. PMID- 9344482 TI - PROTOANEMONIN-INDUCED CYTOTOXIC EFFECTS IN EUGLENA GRACILIS AB - The changes that protoanemonin, an antimicrobial agent of natural origin, brought about in the alga Euglena gracilis were studied with light and electron microscopy and with flow cytometry. The compound proved lethal for the alga at a dose of 5x10(-5)m. At the sublethal dose of 3.5x10(-5)m it caused: marked inhibition of growth; increase in the average cell volume; inhibition of cytokinesis and induction of coenocytic organisms; loss of the flagellum and stigma. Most nuclei were arrested in the G2/M phase of the cellular cycle. The chloroplasts were still well organized, but there was a conspicuous decrease in carotenoids and chlorophylls a and bwith a corresponding increase in pheophytins. The multifarious alterations seen in Euglena are discussed on the basis of a possible interaction between this lactone and several enzymatic systems.Copyright 1997 Academic Press Limited Copyright 1997Academic Press Limited PMID- 9344481 TI - Categorical speech perception in cerebellar disorders. AB - Keele and Ivry (1991) considered the cerebellum an "internal clock" responsible for temporal computations both in the motor and in the perceptual domain. These authors, therefore, expected that the processing of durational parameters of the perceived acoustic speech signal such as voice onset time (VOT) depends upon the cerebellum as well. However, a preliminary investigation of Ivry and Gopal (1992) revealed unimpaired phoneme-boundary effects in cerebellar patients along a continuum of monosyllabic stimuli with systematically varied VOT (/ba/-/pa/). Since the energy of the aspiration noise provides additional cues for the discrimination of voiced and voiceless stops, the present study used a series of disyllabic stimuli differing in a purely durational parameter. Both controls and patients with unilateral cerebellar lesion identified the endpoints of this continuum in nearly all instances as the counterparts of a minimal pair (Boten, /bo:tn/, "messengers" versus Boden, /bo:dn/, "floor"). Subjects with bilateral pathology of the cerebellum, in contrast, did not show a comparable phoneme boundary effect. Our data corroborate the hypothesis of the cerebellum as an internal clock and implicate a role of this structure in speech perception. PMID- 9344483 TI - PPRAS1PPRAS2 AND PPRAP1 GENES, MEMBERS OF A RAS GENE FAMILY FROM THE TRUE SLIME MOLD PHYSARUM POLYCEPHALUM ARE DEVELOPMENTALLY REGULATED AB - The expression patterns of two true ras genes, Ppras1 and Ppras2and one rap gene, Pprap1were examined in four Physarum polycephalum developmental stages: uninucleate amoebae, plasmodia (multinucleate syncytia), spherules (a vegetative, dormant stage) and fruiting bodies. Ppras1 and Pprap1 are expressed in all stages examined with the maximum levels of their transcripts in amoebae and fruiting bodies, respectively, and the minimum levels in plasmodia, whereas the Ppras2 transcript is only detectable in amoebae and fruiting bodies. The results obtained indicate that P. polycephalum is an organism possessing a developmentally regulated ras gene family and presents a convenient system to study the role of ras/rap genes in control of growth and differentiation of lower eukaryotic organisms.Copyright 1997 Academic Press Limited Copyright 1997Academic Press Limited PMID- 9344484 TI - The effect of retrieval cues on visual preferences and memory in infancy: evidence for a four-phase attention function. AB - Bahrick and Pickens (1995) proposed a four-phase model of infant attention, suggesting that recent memories are expressed as a visual preference for novelty, intermediate memories as a null preference, and remote memories as a preference for familiarity. The present study tested a hypothesis generated from this model that a retrieval cue would increase memory accessibility and shift visual preferences toward greater novelty to resemble more recent memories. Results confirmed our predictions. After retention intervals associated with remote memory, previously observed familiarity preferences shifted to null preferences, whereas after a retention interval associated with intermediate memory, the previously observed null preference shifted to a novelty preference. Further, a second experiment found that increasing the exposure to the retrieval cue could shift the familiarity preference to a novelty preference. These findings support the four-phase model of infant attention and suggest that novelty, null, and familiarity preferences lie along a continuum and shift as a function of memory accessibility. PMID- 9344485 TI - Working memory and children's mental addition. AB - Two experiments investigated the extent to which children's mental arithmetic is constrained by working memory rather than their arithmetical competence. A span procedure was used to measure the limit on English- and German-speaking children's ability to add together pairs of multidigit numbers. The children's ages ranged from 7 years 7 months to 11 years 5 months. Spans for mental addition were higher when the numbers to be added were visible throughout calculation than when they were not, consistent with a working memory constraint. Variation in addition span with children's age and with difficulty of the arithmetical operations approximated to a linear function of the speed of adding integers. A similar speed/span relationship has previously been observed for counting span, an artificial task designed to load working memory by combining separate processing and storage subtasks. We conclude that the natural task of mental addition, which combines processing and storage as intrinsic components, reflects working memory in a similar way. Results were remarkably similar both between cultures and across age groups, consistent with the notion of working memory as a general-purpose resource with dynamics that are indifferent to the detailed nature of operations. PMID- 9344486 TI - Children's affective responses, cognitive appraisals, and coping strategies in response to the negative affect of parents and peers. AB - Although research has linked difficulties in parent mood functioning to developmental problems in children, little work has examined why such a link occurs. Following current social-cognitive perspectives on children's cognitive appraisals to negative parent affect, it was hypothesized that children would show more intense affective responses, less confidence, and less active coping strategies in response to parent, as opposed to peer, negative affect. In the current study, young children (N = 39) were read experimental vignettes portraying peers and parents in either happy, sad, or angry emotional states. Children were then interviewed about their affective responses, cognitive appraisals, and coping strategies to each vignette. Beyond experiencing more negative affective responses to parent, compared to peer negative affect, children felt they could do little to help themselves when faced with paternal distress and frequently indicated they would engage in avoidant coping strategies (e.g., hiding) to make themselves feel better when confronted with parent sadness. This study has implications for more industrious future research, as well as intervention projects that involve assisting children who live in households marked by high levels of negative adult affect. PMID- 9344487 TI - Phonological skill and articulation time independently contribute to the development of memory span. AB - One hundred twenty 6- to 10-year-olds were administered three measures of speed of processing, three memory span tasks, three tasks assessing phonological skill, and three articulation tasks assessing the speed with which they could say familiar stimuli aloud. Regression analyses revealed that performance on the span tasks was predicted by performance on the phonology and articulation tasks but not by age or performance on the processing speed tasks. Results are discussed in terms of the processes responsible for age-related change in memory span. PMID- 9344488 TI - Dissociation between features and feature relations in infant memory: effects of memory load. AB - Four experiments examined the effects of the number of features and feature relations on learning and long-term memory in infants. Three-month-olds learned to activate a mobile composed of either two or three kinds of blocks that differed in color, the figures displayed on them, and the figures' colors. Twenty four hours later, infants trained with two objects discriminated feature recombinations but those trained with three objects did not. Even infants trained with three objects discriminated novel features, however, indicating that they remembered the individual features but not the relations among them. A subsequent experiment revealed that this dissociation between features and relations was induced by differential accessibility to memory rather than an encoding failure. We conclude that the size of the memory load selectively constrains infants' long term memory for relational information. These results suggest that in infancy, as in adulthood, features and relations are psychologically distinct and that memory organization parallels the organization of perceptual processing. PMID- 9344489 TI - Children's analogical reasoning about natural phenomena. AB - This report investigates children's analogical reasoning in a physics task, using an analogy generated by the children rather than by the experimenter. A total of 127 elementary school children took part in three related studies. Children learned to predict the behavior of a balance scale. Later, they were asked to solve a force interaction problem. Two versions of the balance scale training were devised: version A suggested an incorrect solution to the target problem (negative analogy), and version B suggested a correct solution to the target problem (positive analogy). In Study 1, 9- to 10-year-olds showed spontaneous transfer in both training conditions. In Study 2, 7-year-olds did not show any transfer in the positive analogy condition. Study 3 revealed that the lack of transfer in younger children was not due to a failure either to notice the analogy or to perform the mapping. Instead, 7-year-olds transferred only selected aspects of the correct solution. PMID- 9344490 TI - Prethymic expression of a transgenic TCR beta chain on a precursor of T-cells. AB - Mice carrying a rearranged TCR Vbeta 8.2 transgene express the Vbeta protein on the vast majority of peripheral T-cells. The bone marrow and peripheral blood, as well as other lymphoid organs of both untreated animals and animals depleted of T cells by neonatal thymectomy and/or injection from birth of monoclonal anti-TCR antibodies, contain a small population of cells that express low levels of the Vbeta transgene product, but no T-cell or other detectable lineage-specific phenotypic markers. When such TG-bearing BM cells are purified and injected directly into the non-TG thymus, they show the phenotypic maturation sequences of intrathymic T-cell development and, subsequently, mature TG-bearing peripheral T cells. However, this population failed to support long-term recovery from lethal irradiation. Both Vbeta 8.2 TG and CD3delta mRNA transcripts are strongly expressed in the cell population, but no CD3gamma, CD3epsilon, CD3zeta, CD4, CD8beta, pre-Talpha, or RAG-1 transcript was detected. The transgene-encoded TCR component is not bound to the cell membrane exclusively by a phosphatidylinositol linkage. The data show that the fully rearranged TCR transgene and transcripts for at least one of the associated CD3 components, CD3delta, can be expressed on a subpopulation of BM and PBL cells that has not passed through the thymus. The phenotypic characteristics of this cell population resemble those described for the earliest thymocyte described by others. The TG protein molecule in this model may provide a specific developmental marker for a prothymocyte lineage subset that lacks pluripotential properties. PMID- 9344491 TI - Role for CD40-mediated activation of c-Rel and maintenance of c-myc RNA levels in mitigating anti-IgM-induced growth arrest. AB - CD40 crosslinking on B cells activates NF-kappaB and stress-activated protein kinase (SAPK) pathways. Since CD40 crosslinking rescues WEHI 231 B cells from anti-IgM-induced apoptosis, those pathways were likely candidates to be involved. Indeed, both signaling cascades predominated in anti-IgM-treated WEHI 231 cells, treated concurrently with anti-CD40 to rescue them from apoptosis. Crosslinking of CD40 activated the NF-kappaB proteins c-Rel and p50, but had no influence on their cytoplasmic steady state level. However, in contrast to-and even in the presence of-anti-IgM-mediated signals, engagement of CD40 resulted in a prolonged nuclear translocation of c-Rel, thereby allowing the formation of active NF kappaB complexes. Consistent with this, the upstream regulatory element of the c myc promoter, known to be regulated by NF-kappaB, was differently regulated after BCR ligation vs BCR plus CD40 crosslinking. The level of c-myc RNA was rapidly downregulated after BCR engagement, but persistent in the presence of CD40 signaling. PMID- 9344492 TI - The lectin jacalin specifically triggers cell signaling in CD4+ T lymphocytes. AB - The lectin jacalin was shown to specifically stimulate CD4(+) lymphocytes. This lectin, which presents a peptide highly similar to a sequence of the HIV external glycoprotein, interacts with CD4 and is able to inhibit in vitro HIV infection. Since jacalin binds also CD8, its mitogenic specificity cannot exclusively be attributed to its interaction with CD4. We therefore hypothesized that the lectin could trigger signals specifically associated with CD4. Here we show that jacalin triggers IL2 gene transcription only in CD4(+) lymphocytes. In parallel, we show that numerous proteins are tyrosine phosphorylated in this cell subset while only a restricted number of them are phosphorylated in CD8(+) cells. Moreover, we show that the tyrosine kinase p56lck, which is associated with both CD4 and CD8, is activated only in CD4(+) lymphocytes, making this lectin a good model for the study of cell signaling triggered in this restricted subpopulation. PMID- 9344493 TI - Inhibition of airway inflammation in rDer f 2-sensitized mice by oral administration of recombinant der f 2. AB - Recombinant Der f 2 (rDer f 2) is a promising new allergen expected to improve the diagnosis and immunotherapy of house dust mite allergy and to further immunological studies. To evaluate the hyposensitizing activity of rDer f 2 to mite allergy, we examined the effect of its oral administration on allergic inflammation in A/J mice immunized with mite allergens. A/J mice immunized with rDer f 2 alone or rDer f 2 + crude mite extract were orally given 0 (control), 0.01, 0.1, or 1 mg/day of rDer f 2 for 4 weeks, followed by an antigen inhalation challenge. Twenty-four hours after rDer f 2 inhalation, control animals experienced severe leukocyte influx into the airway. The infiltrating cells were mainly neutrophils, with some eosinophils and lymphocytes. The concentrations of IL4, IFNgamma, and soluble ICAM-1 in the bronchial alveolar lavage fluid increased twofold compared with values before rDer f 2 inhalation. In contrast, inflammation was significantly suppressed in mice given 1 mg/day of rDer f 2 orally for 4 weeks and partially suppressed in those fed 0.1 mg/day of the antigen. Plasma anti-rDer f 2 antibody levels were unchanged by oral rDer f 2 treatment. Mite extract inhalation challenge provoked neutophilia in rDer f 2 + mite-sensitized control mice, and this allergic reaction tended to decrease in sensitized mice fed 1 mg/day of rDer f 2 orally for 4 weeks. We conclude that rDer f 2 has hyposensitizing activities and may be useful in immunotherapy for house dust mite allergy. PMID- 9344494 TI - Cocaine inhibits human endothelial cell IL-8 production: the role of transforming growth factor-beta. AB - Cocaine use is associated with modulation of a broad range of biological functions including the capacity to influence cytokine production in murine and human immunoeffector cells. Little is known, however, regarding the effects of cocaine on endothelial cell cytokine production. Because the vascular endothelium actively participates in acute and chronic inflammatory responses and interleukin 8 (IL-8) is one of the key cytokines involved in the inflammatory process, modification of the production of IL-8 by vascular endothelial cells may interfere with the host response to infection or tissue injury. We investigated the effect of cocaine on endothelial cell IL-8 production. Conditioned supernatant from EA.hy 926 cells were evaluated by ELISA following in vitro cocaine exposure. Cocaine decreased IL-8 production in a dose-responsive manner, and this reduction correlated with down-regulation of IL-8 mRNA expression. Cocaine also increased the production of TGF-beta by EA.hy 926 cells and anti-TGF beta abrogated the cocaine-mediated decrement of IL-8 production, indicating that cocaine down-regulates endothelial IL-8 production by increasing TGF-beta. Our findings suggest that the immunomodulatory effects of cocaine may be mediated, in part, by modification of endothelial-derived cytokine production. PMID- 9344495 TI - Expression of somatostatin receptor subtype 2 mRNA in human lymphoid cells. AB - We analyzed the mRNA expression of somatostatin receptor subtypes 1 to 5 (SSTR1 5) in human lymphoid cell lines, human peripheral blood lymphocytes (PBL), and human lymphatic leukemia cells, using the reverse transcription-polymerase chain reaction method. In human lymphoid cell lines, SSTR2 mRNA expression was clearly detectable, and there was no evidence of SSTR1 mRNA expression. SSTR2 mRNA was barely detectable in PBL from healthy individuals but was clearly detectable in EB virus-transformed lymphocytes. Lymphocytes from some of the leukemic patients showed elevated SSTR2 mRNA expression. SSTR2 mRNA expression in PBL was upregulated upon stimulation by PHA. SSTR3 mRNA was also observed in all the cell lines examined, although in one cell line, the expression was weak. Some cell lines showed little or no SSTR4 or 5 mRNA expression. The expression pattern of SSTR2 mRNA suggests that this receptor may have some important roles in lymphocyte activation, development, and/or tumorgenesis. PMID- 9344496 TI - A killer cell protective antigen expressed by MHC-unrestricted killer hybridomas. AB - We have established MHC-unrestricted killer hybridoma cell lines. A monoclonal antibody (mAb K2D) described in this study destroyed some of these killer hybridoma cell lines when added to their cultures. This destruction of cells required cell-to-cell contact, indicating that the killer hybridoma cells were killed by others of the same killer hybridoma(fratricide). Based on the facts that the killer hybridoma used did not express FcgammaRIII, and that F(ab')2 fragments of mAb K2D also induced the cell destruction, antibody-dependent cellular cytotoxicity (ADCC) was excluded from a possible mechanism underlying this fratricidal killing. The molecular characteristics of the antigen recognized by mAb K2D and the results obtained by sequential immunoprecipitation led us to conclude that mAb K2D recognizes the Ly-49G2 molecule already described as a member of the Ly-49 family of NK cell inhibitory receptors. Thus, these results suggest that the killer hybridoma cells are protected by the inhibitory signals delivered by Ly-49G2 from killing each other. In addition, we found that mAb K2D recognizes an additional Ly-49-like antigen other than Ly-49G2 on the killer hybridoma, which may reflect the complexity of the Ly-49 family proteins. PMID- 9344497 TI - Differential effects of LPS and CD40 ligand stimulations on the induction of IL 12 production by dendritic cells and macrophages. AB - In this experiment, we have examined the IL-12 production of murine macrophages (Mphi) and dendritic cells (DC) in response to LPS and CD40 ligand (CD40L) stimulations. Splenic Mphi and DC were purified by sorting on the basis of Mac-1 and CD11c expression and stimulated with LPS or Chinese hamster ovary cells expressing CD40L. The results showed that the ligation of CD40 induced the enhancement of IL-12 p40 mRNA accumulation and IL-12 production in DC as well as in Mphi; however, neither the accumulation of IL-12 p40 mRNA nor the production of bioactive IL-12 was detected in DC stimulated with various concentrations of LPS, although Mphi produced IL-12 on LPS stimulation. There was a remarkable difference in the expression of LPS receptor CD14 between Mphi and DC. Mphi evidently expressed CD14 but the CD14 expression of DC was quite low. Possible mechanisms of the failure in IL-12 production of DC are discussed. PMID- 9344498 TI - Role of interferon-gamma in the priming of decidual macrophages for nitric oxide production and early pregnancy loss. AB - We have previously shown that both priming and triggering signals were needed for nitric oxide production by decidual macrophages and that nitric oxide was responsible for embryo wastage. In this study, we investigated the role of IFN gamma as the primary signal for macrophage activation in early embryo loss. IFN gamma-deficient (GKO) and heterozygous F1 control mice were injected with lipopolysaccharide (LPS) at day 7 of gestation. The results showed that the GKO mice were more resistant to LPS-induced embryo loss than the wild type. This suggested that IFN-gamma was needed for LPS-induced embryo resorption and that decidual macrophages from pregnant GKO mice were not primed and could not be activated when given LPS. Further, the results showed that IFN-gamma mRNA was simultaneously expressed in the same embryos that also expressed mRNA markers for macrophage activation (TNF-alpha and iNOS), indicating that macrophage activation could be a consequence of IFN-gamma production. Similarly, we investigated the role of IL-12 as a switch cytokine capable of eliciting TH1-associated cytokine production including IFN-gamma. The results showed that IL-12 mRNA expression was correlated with IFN-gamma expression and macrophage activation. In this in vivo study, we showed for the first time that spontaneously increased decidual IFN gamma expression is detrimental to embryo survival. PMID- 9344499 TI - Ligation of CD2 provides a strong helper signal for the production of the type 2 cytokines interleukin-4 and -5 by memory T cells. AB - In this study, we investigated the efficiency of costimulatory signaling provided by anti-CD2 monoclonal antibodies (mAbs) for the production of type 1 (IL-2 and IFN-gamma) and type 2 (IL-4, IL-5, and IL-10) cytokines by human T cells. We cultured purified human T cells (freshly isolated from blood) with immobilized anti-CD3 mAb, either alone or in combination with anti-CD28 or anti-CD2 mAbs. When compared with the standard costimulatory signal anti-CD28, anti-CD2 mAbs (9 1 plus 9.6) were shown to be more potent costimulators of IL-4 production and to have similar activity for IL-5 production, but to be less potent for costimulation of IL-2, IL-10, and IFN-gamma production. IL-4 production was completely inhibited by cyclosporin A or by blocking IL-2 activity and its receptor. IL-4 and IL-5 were produced by CD45RO+ T cells but not by CD45RA+ T cells, indicating that anti-CD2 in this system costimulated type 2 cytokine production by differentiated memory cells. In the presence of IL-12, the cytokine profile was shifted to high IFN-gamma and IL-10 production and IL-4 and IL-5 production were slightly inhibited. Our data thus suggest that CD2 ligation plays an important role in the upregulation of Th2-like T cell activity (especially IL 4 production), but they also show that this effect is strongly modulated by IL 12, resulting in predominant IL-10 and IFN-gamma production instead. PMID- 9344500 TI - Prostaglandin E2 and IL-4 provide naive CD4+ T cells with distinct inhibitory signals for the priming of IFN-gamma production. AB - We investigated the effects of prostaglandin E2 and IL-4 on the acquisition of cytokine-producing ability by naive CD4(+) T cells in human umbilical cord blood. The presence of PGE2 or IL-4 at primary stimulation inhibited the production of IFN-gamma at secondary stimulation, and the combination of these stimuli resulted in cooperative effects. During primary stimulation with anti-CD3, the intracellular cAMP level was elevated in PGE2-treated cells but not in IL-4 treated or control cells. The signal provided by PGE2, but not by IL-4, was inhibited with RpcAMP, indicating that it was mediated by cAMP. After differentiation into Th2-like cells, cAMP levels in PGE2- and IL-4-treated cells were not different. Our results suggest that both PGE2 and IL-4 play important roles with distinct mechanisms in inhibiting the priming of IFN-gamma production of naive CD4(+) T cells. PMID- 9344501 TI - Activation-induced T cell death: resistance or susceptibility correlate with cell surface fas ligand expression and T helper phenotype. AB - Activated T cells undergo apoptosis when the Fas-antigen (Apo-1, CD95) is ligated by Fas ligand molecules (FasL) or agonistic anti-Fas antibodies. Restimulation of T lymphocytes via the TCR/CD3 complex induces activation-induced cell death (AICD). AICD and Fas-induced cell death are causally related since TCR-induced AICD at least in part depends on Fas/FasL interactions. Thus, restimulation of T cells leads to FasL gene transcription and surface expression. Membrane-bound or secreted FasL molecules then bind to Fas receptors on the same cell or on a neighbor cell to trigger the death signaling cascade. We have compared Fas mediated apoptosis and AICD in a panel of human T cell clones. While all clones were killed by anti-Fas mAb, several clones were resistant to AICD triggered by anti-TCR/CD3 mAb or superantigen. The pattern of TCR-induced protein tyrosine phosphorylation was comparable in AICD-resistant and -susceptible clones, as was the induction of FasL mRNA. However, significant differences were observed at the level of FasL surface expression which was induced in AICD-susceptible but not in AICD-resistant clones. Cytokine profiles of CD3-stimulated clone cells support the recent observations that AICD sensitivity is restricted to the Th1 subset. However, AICD-resistance is not only associated with the classical Th2 phenotype. PMID- 9344502 TI - Semi-quantitative polymerase chain reaction analysis of cytokine and cytokine receptor gene expression during thymic ontogeny. AB - Semi-quantitative, polymerase chain reaction (PCR) is used to uncover the patterns of cytokine transcription in the mouse thymus from day 14 to day 20 of gestation, a time period which includes many of the important events in thymic ontogeny. Interleukin 4 (IL-4), IL-7 and interferon gamma (IFN-gamma) mRNA is abundant from fetal day (Fd) 14-16, corresponding with the period of rapid proliferation of immature thymocytes in vivo. As the level of mRNA for these cytokines diminishes, the induction and increased expression of IL-3 and IL-2 occurs. The transcription of these cytokines correlates temporally with the period of proliferation-dependent phenotypic differentiation between Fd 16 and 20. The thymic epithelium (TE)-derived cytokines including IL-1alpha, IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) begin to be transcribed between Fd 14-15 and show peak mRNA abundance from Fd 16-20. IL-5, tumour necrosis factor alpha (TNF-alpha) and LT (lymphotoxin or TNF-beta) constitute a fourth group of cytokines, along with the IL-4 receptor (IL-4R), which are transcribed at an even level throughout the fetal period. The IL-2 receptor beta chain (IL-2Rbeta) and IL-10 show abundant mRNA from Fd 14-20 and have a peak level of mRNA content on Fd 16. Taken together, these studies uncover complex, overlapping patterns of cytokine gene expression. The mRNA abundance and pattern of expression of each cytokine or cytokine receptor may indicate the relative contribution that it makes to different stages of fetal thymic ontogeny. PMID- 9344503 TI - Modulation by lipopolysaccharide of inflammatory cytokine production by two T cell lines. AB - In this study, the modulation of inflammatory cytokine production by lipopolysaccharide (LPS) in two T cell lines, RL-male-1 and L5178Y-ML was examined. Both cell lines produced interleukin 1 (IL-1) activity in response to LPS, which was largely independent of the presence of serum. The IL-1 activity induced was neutralized by an anti-IL-1beta antibody, but not by an anti-IL 1alpha antibody. Induction of IL-1alpha and beta was also confirmed by polymerase chain reaction of reverse-transcribed mRNA (RT-PCR), although in RL-male-1 cells, IL-1alpha mRNA was constitutively expressed and somewhat enhanced by LPS. RT-PCR analysis revealed that these cell lines also upregulated tumour necrosis factor alpha (TNF-alpha) and IL-1 receptor antagonist mRNAs in response to LPS, although RL-male-1 cells expressed TNF-alpha mRNA constitutively. Flow cytometric analysis showed that, although these cells expressed Thy-1 antigen, they hardly expressed CD14 and gammadelta T cell receptor. In conclusion, LPS modulated inflammatory cytokine production in these T cell lines. PMID- 9344505 TI - Endothelial cell proliferation regulated by cytokines modulates thrombospondin-1 secretion into the subendothelium. AB - Endothelial cells activation and proliferation are induced by cytokines and modulated by adhesive molecules of the extracellular matrix. In a previous study, we showed that IL-1beta and TNF-alpha inhibited thrombospondin-1 (TSP) secretion by human umbilical vein endothelial cells. This work was carried on by studying the effects of other cytokines [interleukin 6 (IL-6), IL-8, basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta)] known to modulate endothelial cell proliferation. We show here that TSP incorporation into the subendothelial matrix is inversely proportional to cell density. Proliferative cytokines such as IL-6 and bFGF inhibit TSP secretion, whereas the anti-proliferative TGF-beta enhances it. IL-8 has no effect either on cell proliferation or TSP secretion. Thus, the modulation of TSP secretion by these cytokines is apparently due to changes in cell proliferation. Therefore, when studying the effects of cytokines on TSP secretion, it is important to correlate the concentration of subendothelial matrix TSP to the cell density. PMID- 9344504 TI - Recombinant human IL-2 is cytotoxic to oligodendrocytes after in vitro self aggregation. AB - Interleukin 2 (IL-2) is a cytokine produced by activated T cells that plays a crucial role in the immune response and exerts multiple functions in different tissues including the nervous system. The present study was carried out to investigate the effect of recombinant human IL-2 (rhIL-2) on the survival of differnet glial cells type and of cortical neurons in culture. The results demonstrate that rhIL-2 is selectively cytotoxic to oligodendrocytes only if preincubated in aqueous solution (1200 U/ml) for at least two days before being added to the culture. Other glial and neuronal cells were unaffected. The cytotoxic effect was temperature- and concentration-dependent occurring only when rhIL-2 was reincubated at 37 degrees C and was dose-dependently neutralized by antibodies raised against IL-2 indicating that an immunoreactive IL-2 like compound is the active principle. Polyacrilamide gel electrophoresis under native (PAGE) and denaturing (SDS-PAGE) conditions and gel filtration analysis of the rhIL-2 incubated solution suggest that rhIL-2 is cytotoxic to oligodendrocytes only after association into soluble high weight molecular aggregates. PMID- 9344506 TI - CD19-selected B lymphocytes synthesize, secrete and migrate in the presence of IL 8. TNF-alpha and gammaIP-10 are also B lymphocyte migratory factors. AB - B lymphocytes are responsible for antigen uptake and presentation, as well as antibody production. These reactions require close cell-to-cell contact between B lymphocytes and monocytes. In this study we demonstrate that interleukin 8 (IL 8), gamma-immune protein 10 (gammaIP-10) and tumour necrosis factor alpha (TNF alpha) all induce a significant chemokinetic response of human B lymphocytes. Among the cytokines tested, rIL-8 was the strongest B lymphocyte migratory factor with a migratory index (MI) of 2.03+/-0.32, (P<0.002), followed by rTNF-alpha (MI=1.89+/-0.17, P<0.001) and rgammaIP-10 (MI=1.63+/-0.17, P<0.001). We did not observe B lymphocyte migration towards rIL-1alpha, rIL-2, rIL-4, rIL-10, interferon gamma (rINF-gamma) or transforming growth factor beta (rTGF-beta). Furthermore, we report that human B lymphocytes have a constitutive IL-8 mRNA expression and protein secretion in vitro. Resting as well as stimulated B lymphocytes secrete on average 1.5 ng IL-8/ml medium/24 h (2x10(6) B lymphocytes). Our data indicate a possible mechanism by which B lymphocytes make contact with other cells, during immuno-inflammatory processes. PMID- 9344508 TI - Beta-amyloid fragment potentiates IL-6 and TNF-alpha secretion by LPS in astrocytes but not in microglia. AB - The effect of a peptide homologous to the biologically active fragment of beta amyloid 25-35 (beta 25-35) was studied on interleukin 6 (IL-6) and tumour necrosis factor (TNF-alpha) secretion induced by lipopolysaccharide (LPS) in primary rat astrocytes and microglia. Twenty-four hour exposure to LPS (50 ng/ml) induced IL-6 and TNF-alpha both in astrocytes and in microglial cells, while the effect of beta 25-35 (50 microM) per se was negligible in both cell types. In microglial cells, the application of beta peptide did not alter the production of either cytokine induced by LPS. However, beta 25-35 strongly amplified the production of both IL-6 and TNF-alpha in astrocytes. These findings confirm the complex interaction between cytokines and amyloidogenesis in Alzheimer's disease and indicate that astrocytes rather than microglia respond to the beta amyloid fragment, suggesting that these cells may be actively involved in cytokine mediated events in AD. PMID- 9344507 TI - Spontaneous and inducible production of leukaemia inhibitory factor by human bone marrow stromal cells. AB - Leukaemia inhibitory factor (LIF) acts on the growth and differentiation of haematopoietic cells. By using a specific enzyme-linked immunosorbent assay for human LIF, we demonstrate that human bone marrow stromal cells produce LIF. LIF synthesis is enhanced in a dose-dependent manner after stimulation with lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMAS). LIF production in response to PMA is PKC-dependent since the two PKC inhibitors sphingosine and staurosporine markedly diminished it. Interleukin 1alpha (IL 1alpha), IL-1beta, IL-3, IL-6, IL-8, tumour necrosis factor (TNF-alpha) and SCF (both at 10 ng/ml) stimulate LIF production. By contrast macrophage colony stimulating factor (M-CSF), granulocyte (G)-CSF, GM-CSF, basic fibroblast growth factor (bFGF), platelet-activating factor (PAF), protaglandin E2 (PGE2), leukotriene B4 (LTB4), and leukotriene C4 (LTC4) did not. These results suggest that bone marrow stromal cells might represent a major source for the cytokine regulated local production of LIF inside human bone marrow. PMID- 9344509 TI - Adverse effects of tumour necrosis factor in cyclophosphamide-treated mice subjected to gut-derived Pseudomonas aeruginosa sepsis. AB - To evaluate the role of tumour necrosis factor (TNF) in gut-derived sepsis, mice were given Pseudomomas aeruginosa strain D4 by bacterial suspension in their drinking water during which time ampicillin (200 mg/kg) was given to disrupt the normal indigenous bacterial flora. Cyclophosphamide was additionally administered to induce bacterial translocation of the P. aeruginosa that had colonized the gastrointestinal tract, and thereby to cause gut-derived sepsis. In this model, TNF-alpha was detected in serum from the next day after the second cyclophosphamide administration, increasing to level of 3 ng/ml in lethal conditions. Average serum TNF-alpha level was significantly higher in mice with bacteraemia than in those without bacteraemia. Treatment with 0.8 microg/kg of recombinant human TNF-alpha (rhTNF-alpha) did not affect the mortality, whereas administration of either 4 and 20 microg/kg of rhTNF-alpha significantly increased the mortality rate in comparison with saline-treated mice. Bacterial counts in liver and blood were significantly higher in 20 microg/kg of rhTNF alpha treated mice than in saline-treated mice. Treatment with murine anti-TNF alpha monoclonal antibody significantly reduced the mortality from septic infection. We conclude that TNF-alpha may facilitate bacterial translocation and causes deterioration of gut-derived sepsis due to P. aeruginosa in mice. PMID- 9344510 TI - TIMP-3 induces cell death by stabilizing TNF-alpha receptors on the surface of human colon carcinoma cells. AB - Matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) regulate the structural integrity of the extracellular matrix (ECM). Constitutive expression of human TIMP-3 in human DLD colon carcinoma cells renewed serum-responses and inhibited tumour formation in nude mice. To elucidate the mechanism of TIMP-3-mediated tumour suppression, we compared parental DLD and TIMP-3 expressing DLD cells (TIMP-3/DLD), finding them to be significantly different. TIMP-3/DLD cultures have fewer mitotic cells, are delayed in G1, and die after serum starvation. TIMP-3/DLD conditioned media activates cell death on fibroblast cells. The cell death induced by serum starvation and conditioned media was inhibited by 70%, in the presence of neutralizing tumour necrosis factor alpha (TNF-alpha) antibody. TIMP-3/DLD whole cell lysate contained p55 TNF alpha receptor, while vector/DLD lysate had p55 TNF-alpha receptor and p46 soluble TNF-alpha inhibitor. Vector/DLD conditioned media had p46, while no soluble TNF-alpha receptor was detected in TIMP-3/DLD conditioned media. In addition, FACS analysis revealed that TIMP-3/DLD cells have more TNF-alpha surface binding sites, suggesting a direct correlation between TIMP-3 expression and surface receptors. The mechanism of tumorigenic reversion induced by TIMP-3 in DLD cells may involve protection of receptors from the proteolytic activity of MMPs. Putative TIMP-3-mediated inhibition of MMPs restores the TNF-alpha p55 signalling pathway and the carcinoma cell is killed by autocrine TNF-alpha. Thus, DLD cells have specific ECM MMPs that cleave cytokines and cytokine receptors. TIMP-3 specifically inhibits MMPs involved in receptor shedding. PMID- 9344511 TI - Protective effects of granulocyte colony-stimulating factor on endotoxin shock in mice with retrovirus-induced immunodeficiency syndrome. AB - Mice with retrovirus-induced murine acquired immunodeficiency syndrome (MAIDS) were hypersensitive to lipopolysaccharide (LPS)-induced lethal shock accompanied by marked elevations of systematic interleukin 1beta (IL-beta) and interferon gamma (IFN-gamma) after LPS challenge. Pretreatment with 10 microg of recombinant human granulocyte colony-stimulating factor (rhG-CSF) protected MAIDS mice from hypersensitivity to LPS-induced lethal shock and this protection was concomitant with suppression of IFN-gamma production. PMID- 9344513 TI - Compact Fiber Optic Dynamic Light Scattering System AB - Fiber optic dynamic light scattering (FODLS) is fast developing as a technique for sizing particles in concentrated dispersions. Fiber optic systems have the potential to be made into online particle size measuring equipment in process industries. Two different laser light sources were used to study monodisperse polystyrene latex dispersion with different particle sizes and volume fractions. The apparent diffusion coefficients were a function of particle size, volume fraction, and signal to noise ratios. The apparent diffusion coefficient data showed that FODLS probes collective diffusion for smaller particles and self diffusion for larger particles. It was observed that for a polydisperse concentrated dispersion, both self and collective diffusion were probed by FODLS. This means that the apparent diffusion coefficient can be used only as a relative quantity. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344512 TI - Tumour necrosis factor alpha promoter gene polymorphism in sarcoidosis. AB - Biallelic polymorphisms in the promoter region of the TNF-alpha gene (TNFA) and in the first intron of the TNF-beta gene (TNFB) have been associated with variation in TNF-alpha production and with susceptibility to severe diseases. Among other functions, TNF-alpha plays a pivotal role in regulatory aspects of granuloma formation and sustenance. In sarcoidosis, a systemic granulomatous disorder of unknown aetiology, the clinical course of the disease has been associated with the patient's individual capacity of spontaneous TNF-alpha production by alveolar macrophages. We determined the TNFA and TNFB polymorphisms in 101 patients with pulmonary sarcoidosis and 216 healthy blood donors. A highly significant shift to the more uncommon TNFA2 allele was found in the Lofgren syndrome patient group, which represents the acute form of the disease with frequent spontaneous remission. The results show that gene frequencies of the TNFA gene variation are significantly different within the clinical forms of sarcoidosis, indicating that genetic predisposition for TNF-alpha production may play a role in the pathogenesis of the disease. PMID- 9344514 TI - A Comparison between Hydrophobically End-Capped Poly(ethylene oxide) with Ether and Urethane Bonds AB - Two models of HEUR (hydrophobically modified ethylene oxide urethane) associative polymers (AP) are compared. The two polymers are polyethyleneoxides simply end capped with dodecyl groups, one through an ether bond H25C12-O-(CH2CH2O)304 C12H25 (AP14, Mw = 13,700) and the other through an urethane bond H25C12-NHCOO (CH2CH2O)304-CONH-C12H25 (AP14NCO, Mw is about the same as for AP14). The results indicate that this subtle difference in polymer architecture dramatically influences the initial association and the clouding of the polymer in water, so that AP14NCO starts to aggregate earlier but has a higher clouding temperature. However, at more elevated polymer contents (above 2-3 wt%) the differences in solution behavior and transport dynamics between the two polymers seems less significant. These conclusions are drawn through fluorescence, dynamic light scattering, NMR self-diffusion, turbidimetry, and viscosity measurements. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344515 TI - Probing the Sol-Gel Conversion in the Tetraethoxysilane/Alcohol/Water System with the Aid of Diffusion-Controlled Fluorescence Quenching AB - The sol-gel conversion in the tetraethoxysilane/alcohol/water system can be probed by means of the diffusion-controlled fluorescence quenching of Py* by Cu2+ ions. In the course of tetraethoxysilane hydrolysis, the viscosity of the system increases, thereby causing the quenching rate to decrease and the mean lifetime of the excited state to elongate dramatically as the system approaches the gel point. Incorporating the equation proposed by Vogelsberger et al. (1992) for the time-dependence of the viscosity of gelating systems into the Smoluchowsky kinetic equation, an equation has been proposed to describe the observed time dependence of the Py* lifetime. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344516 TI - Interaction of o-Aminophenol and o-Nitrophenol with Copper, Zinc, Molybdenum, and Chromium Ferrocyanides AB - Removal of o-aminophenol and o-nitrophenol from aqueous solution through adsorption on copper zinc, molybdenum, and chromium ferrocyanides were studied in pH range 2-10 at 27°C. At pH 7.0 o-nitrophenol adsorbed more than o aminophenol on all the metal ferrocyanides studied. The Langmuir type of adsorption is followed in the concentration range of 10(-3) to 10(-4) M of o aminophenol and o-nitrophenol solutions. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344517 TI - Formation of Surface Active Gelatin by Covalent Attachment of Hydrophobic Chains AB - Surface active gelatin was formed by covalent attachment of hydrophobic groups to gelatin molecules. The modification was carried out at various degrees of attachment and with various chain lengths. These modified gelatins (MGs) were synthesized in dry DMSO by a simple and rapid method. The new method leads to high yields and allows high degrees of modification. The MGs, which have various hydrophobicities, have better surface activity than the native gelatin, as determined by surface tension reduction. The surface tension reduction is correlated to the hydrophobicity of the modified molecule, which was determined by a fluorescent probe. It appears that both the increase in the number of the hydrophobic groups and the increase in the chain lengths lead to decreased surface tension. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344518 TI - Calculation of the Static Permittivity of Suspensions from the Stored Energy AB - It is shown that the static permittivity of suspensions of charged particles in electrolyte solution can be deduced from the field-induced change of the Gibbs free energy stored outside the double layer. This energy is related to the changes of the electrolyte concentration around the particle and is obtained solving a purely static problem. The method is first verified deducing well-known results corresponding to suspensions of spherical particles. It is then used to calculate the static permittivity of suspensions of spheroidal particles, leading to a new result that cannot be analytically obtained using the classical approach. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344519 TI - Adsorption of Polycations on Clays: A Comparative in situ Study Using 133Cs and 23Na Solution Phase NMR AB - 23Na solution phase NMR has been evaluated as an in situ probe to study the adsorption of tetramethylammonium (TMA+) and two polycations, FL17 ([(Me2NCH2CHOHCH2)n]n+Cln) and Magnafloc 1697, ([(CH2CHCH2N(Me)2CH2&Cmacr;HCH2)n]n+Cln), onto clays in aqueous suspensions containing 2.5 mass% low iron Texas bentonite. The NMR data shows the effectiveness of the organocations at displacing Na+ from the bentonite surface. This information has been correlated with that obtained from particle-size and electrophoretic measurements in aqueous solution, together with information from adsorption isotherms. These results have been compared to those obtained in parallel studies using 133Cs solution phase NMR. FL17 and 1697 both exhibited high affinity adsorption isotherms on Na+- and Cs+-clay, whereas the adsorption of TMA+, which represents the cationic portion of the polymers was of lower affinity. Na+-bentonite adsorbed almost twice the amount of polycation required to fulfill the cation-exchange capacity (CEC) of the bentonite. The electrophoretic and particle size data indicated significant differences in the size of the polycation/clay flocs and the amount of polymer adsorbed on the external faces of the flocs in the presence of Na+- and Cs+-exchange ions. Correlation of this data with the NMR results suggests that the Na+ bentonite/polycation flocs are large, of low density, and that the polycation is concentrated in the interior while the Na+-ions occupy exchange sites on the external faces. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344520 TI - A Physicochemical Study of Polypyrrole-Silica Nanocomposites by Inverse Gas Chromatography AB - Three polypyrrole-silica nanocomposites were characterized by inverse gas chromatography (IGC) at 60 and 80degreesC using both linear and branched alkane probes. The IGC data for these materials were compared with those obtained for the ultrafine silica sol and polypyrrole (PPy) bulk powder reference materials. The London components of the surface energies (gammasd) of the nanocomposites were in the range 115-225 mJ m-2 at 60degreesC. These values are much higher than those obtained for either the polypyrrole powders or the colloidal silica sol, which suggests that the nanocomposites have significant microporosity. The gammasd values for the nanocomposites decrease in the order chloride-doped PPy > sulfate-doped PPy > tosylate-doped PPy, which is the same trend as that found for the heat of adsorption (DeltaHa) of a given n-alkane. DeltaHa values of the linear and branched alkanes are up to three times higher than the heats of vaporization, which confirms that the nanocomposites (and, to a lesser extent, the polypyrrole powders) have high adsorption capacities for these solutes. For a series of alkane isomers (e.g., C7H16 or C8H18), the difference between the DeltaHa values for the nanocomposites and the corresponding polypyrrole powders decreased as the degree of branching for these probes was increased. These observations suggest that the more sterically hindered probes are excluded from the microporous interior of the nanocomposites. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344521 TI - The Effect of Neutral Polymer and Nonionic Surfactant Adsorption on the Electroacoustic Signals of Colloidal Silica AB - The effects of adsorbed neutral polymer and nonionic surfactant on the electroacoustic signals of silica particles have been determined. The dynamic mobility of the particles was measured using an AcoustoSizer (Matec Applied Sciences) during the addition of poly(vinyl alcohol) (PVA) and nonyl phenol ethoxylate (C9phiEN) to the system. Large changes in the dynamic mobility of the coated particles were observed. A theoretical treatment was developed to describe the electroacoustic behavior of an elastic gel layer of uncharged adsorbed polymer and provided a reasonable fit to the experimental dynamic mobility data. This analysis provided information about the thickness and elasticity of the adsorbed layers. The adsorbed layer thickness for PVA compared well with results of other techniques reported in the literature and appeared to be influenced by changes in pH. C9phiEN adsorbed in a relatively dense layer. For both PVA and C9phiEN, the thickness of the inner adsorbed layer remained approximately constant, while the outer layer became thicker with increasing concentration and molecular weight. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344522 TI - Adsorption of Dextrin at Mineral/Water Interface AB - The adsorption mechanism of dextrin on aqueous minerals such as fluorite, apatite, galena, magnetite, gamma-alumina, and graphite was studied by adsorption experiments, zeta potential measurements, and FT-IR studies. Depending on the nature of the mineral surface, dextrin was found to interact in three different ways viz. by chemisorption, physisorption, or hydrophobic-hydrophobic interaction. The adsorption density of dextrin was found to be pH dependent. Maximum adsorption of dextrin was obtained around the pH at which the mineral surface is highly hydroxylated. The mechanism of dextrin interaction with the surface metal hydroxy sites, ( identical withMeOH), was found to proceed via chemical complexation. A linear relationship was observed between the adsorption density of dextrin and the pH of maximum surface hydroxylation. Zeta potential measurements have indicated the possibility of dextrin adsorption by electrostatic interaction under the conditions where mineral surface and dextrin are oppositely charged. Furthermore dextrin was found to adsorb on hydrophobic minerals such as graphite by hydrophobic-hydrophobic interaction. However, the magnitude of adsorption by electrostatic and hydrophobic interaction was found to be very marginal compared to that of chemical complexation. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344524 TI - Adsorption of Benzoic Compounds onto Stainless Steel Particles AB - Equilibrium experiment was conducted to investigate the factors determining the adsorption of benzoic acid (BA) and its derivatives, m- and p-hydroxy BA, onto SUS316L stainless steel particles of 8-10 &mgr;m diameter and under 100 mesh. Adsorption isotherms of these benzoic compounds were determined in the presence of 0.05 M NaCl at pH 4 and 30°C. The adsorptions of the these compounds were described well by a Langmuirian model for both adsorbents. When the maximum number of the benzoic compound adsorption sites was expressed on the basis of unit surface area (N, mol/m2), the N values were relatively constant, while the greatest value of the affinity (K, ml/&mgr;mol) was obtained for p-hydroxy BA, although its value was in the same range as that of the other two adsorbates. Diffuse-reflectance Fourier transform infrared spectra of the fine adsorbent (8 10 &mgr;m diameter) after equilibration suggest that the adsorption mainly takes place through the carboxyl group of the adsorbate-stainless steel surface interaction for all adsorbates, whereas concomitant interaction occurs in part with participation of the phenolic hydroxyl group for p-OH BA adsorbate, accounting for the difference in adsorption properties. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344523 TI - Surface Chemical Analysis and Electrokinetic Properties of Synthetic Spherical Mixed Zinc-Cadmium Sulfides AB - In this work, we analyze the surface and bulk chemical composition, as well as the crystal structure, of colloidal spherical particles of Zn-Cd mixed sulfides of different Zn/Cd ratios. The particles were obtained by precipitation from solution according to the method described by Wilhelmy and Matijevic [Colloids Surfaces 16, 1 (1985)]. Transmision electron microscopy of the particles show that their average diameter ranges from 50-60 nm (when the synthesis is carried out at 50degreesC) up to 150-200 nm (for a temperature of 70degreesC). Atomic absorption analysis of the twelve samples obtained indicated that the bulk Zn/Cd ratio increases with aging temperature; the same behavior is found when the concentration of Cd(NO3)2 used in the synthesis is decreased. Similarly, the bulk proportion of Zn in the particles is higher the longer the growth time. EDX microanalysis was also performed on all the samples; although this technique is not a bulk (but rather surface) analytical tool, the fact that it gives information down to a depth of approximately 500 A from the surface makes the type of information obtained with EDX comparable to atomic absorption. Although the overall Zn/Cd trends are reproduced by EDX data, these are not as sensitive as atomic absorption. The surface composition of three selected samples (M3, 50 min growth time, 50degreesC, 0.52 mM Cd2+ in the growing solution; M8, 100 min, 60degreesC, 0.52 mM; M12, 100 min, 70degreesC, 0.52 mM) was determined by XPS spectra od Cd 3d5/2, Zn 2p3/2, and O 1s electrons, for the three samples. The sequence of variation of the Zn/Cd ratio of M3, M8, and M12 particles agrees qualitatively with that found by atomic absorption or EDX; the fact that no detectable Cd is found in sample M12 suggests that the particles have a nonhomogeneous composition that changes from the core to the surface layer. The analysis of O 1s electrons allows to reach the conclusion that the surface oxidation changes in the order M3 > M8 > M12, i.e., the particles are more oxidized the larger the amount of cadmium on their surface. This is confirmed by electric conductivity determinations in aqueous suspensions of the samples, both in the presence of natural light and in the dark, as a function of time. These data, together with crystal structure determinations by XRD, suggest that, when the growth temperature is 50-60degreesC, the particles contain a ZnS (sphalerite) nucleus covered by a layer of mixed, hexagonal Zn-CdS and a surface layer of cubic ZnS. When the aging temperature is 70degreesC, the ZnS core is surrounded by a shell containing cubic ZnS and amorphous CdS. The surface electrical properties of the particles in aqueous suspensions were analyzed by electrophoresis: the effect of pH on the electrophoretic mobility, and in particular the pH value at which the mobility is zero (isoelectric point or pHiep) confirms the conclusions obtained from our previous surface chemical analysis concerning the surface oxidation of the particles. The effect of lattice ions (Zn2+, Cd2+, S2-) in solution on the mobility (and hence on the surface charge) of the particles is very significant: the latter ions are able to find easily their way to the surface of the sulfides and change to a large extent the overall pH-dependence of the mobility and specifically the values of the pHiep. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344526 TI - Spatial Distribution of Chemical Components in Aerosol Particles as Determined from Secondary Electron Yield Measurements: Implications for Mechanisms of Multicomponent Aerosol Crystallization AB - Secondary electron yield measurements were used to determine the distribution of chemical components in multicomponent aerosol particles formed by crystallization from aqueous solution droplets. Yield measurements were made by measuring the charge acquired by a beam of particles as they passed through an electron beam (100-600 eV energy) inside a high-vacuum apparatus. Yields were sufficiently different for certain compounds that measurements made on two-component particles could be used to obtain information on the spatial distribution of components. Variations in chemical composition as a function of depth beneath the particle surface were ascertained from the energy dependence of the measured yields, since the electron penetration depth, and therefore the probe depth, increases with electron energy. The results for mixed NaCl-NH4Cl particles indicate that the distribution of components within these particles is relatively homogeneous, while measurements made on mixed NaCl-NaNO3 particles are indicative of particles having a heterogeneous core-shell morphology. Results for mixed Na2SO4-(NH4)2SO4 particles are ambiguous, probably because of the complexity of the phase diagram of this system. It appears that the mechanism by which aerosol particles crystallize, and therefore the resulting distribution of chemical components within particles, is strongly dependent on particle composition and environmental variables. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344525 TI - Low-Frequency Dielectric Dispersion from Ion Permeability of Membranes AB - Low-frequency dielectric behavior of membrane/electrolyte systems was studied with artificial membranes made from thin plastic films and NaCl electrolyte of various concentrations. A low-frequency dielectric dispersion or relaxation, which is associated with membrane pores and membrane permeability, was observed and investigated. A qualitative explanation is proposed. Membrane structure characteristics, such as thickness, pore size, and number, were found to affect significantly the low-frequency dielectric dispersion, which indicates some potential applications for low-frequency dielectric measurements in membrane- or tissue-related areas. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344527 TI - Mercury Penetration and Snap-off in Lenticular Pores AB - During mercury porosimetry experiments two of the most basic mechanisms responsible for mercury intrusion (drainage) into and retraction (imbibition) from porous media are meniscus intrusion into constrictions and thread snap-off in narrow pores, respectively. The present work consists of an experimental and theoretical study of these phenomena in model lenticular capillaries, that is, capillaries with lens-shaped cross section. The reason for this apparently strange choice is that several porous media of interest, such as sandstone reservoir rocks (and also model pore networks etched in glass plates and used in numerous experimental studies of multiphase flow in porous media) have pores of such type. The critical pressures for mercury intrusion and snap-off in lenticular pores are measured experimentally and expressed in dimensionless form as functions of the pore aspect ratio (ratio of pore width to pore depth) and the contact angle. Analytical mathematical relationships are also developed for the calculation of these critical pressures. In the case of mercury penetration, very good agreement between experiment and theory is observed over the whole region of pore width to pore depth aspect ratio. In the case of mercury snap-off, very good agreement is observed for small and medium values of pore aspect ratio. A sizable discrepancy in the case of snap-off is observed in pores of large aspect ratio, and this is caused by the considerable deviation (in this case) of the pore shape of the experimental models from the lenticular one used in the theoretical calculations, as well as by the strong effect of the pressure of residual air on the values of measured pressures. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344528 TI - Surface Element Integration: A Novel Technique for Evaluation of DLVO Interaction between a Particle and a Flat Plate AB - A novel method, the surface element integration (SEI), is developed to determine the van der Waals and electrostatic double layer interactions between a particle and an infinite flat plate from the corresponding interactions per unit area between two infinite flat plates. Comparison with the Hamaker expression for nonretarded van der Waals interaction reveals that the new technique gives the exact interaction energy between a spherical particle and a flat plate. Available analytical expressions for the electrostatic double layer interaction energy between two infinite flat plates, based on the linearized Poisson-Boltzmann equation, are used in SEI to obtain the sphere-flat plate interaction energy. These sphere-flat plate interaction energies determined using SEI are compared with the corresponding interaction energies obtained from a detailed numerical solution of the Poisson-Boltzmann equation based on finite element analysis. The comparisons reveal that SEI scales the flat plate interaction to the corresponding sphere-flat plate geometry exactly, while the scaling based on the conventional Derjaguin approximation technique grossly overpredicts the interaction energy for small particles and low electrolyte concentrations. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344529 TI - Surface-Enhanced Raman Spectroscopic Study of Isomeric Formylthiophenes in Silver Colloid AB - Surface enhanced Raman (SER) spectra of 2- and 3-formylthiophenes (2FT and 3FT) in silver colloids are compared with their normal Raman spectra in bulk and in acetonitrile solution. Experimental results indicate adsorption of 2FT on metal surface through the sulfur and the oxygen atoms with the ring plane vertical to the metal surface, whereas the ring plane of the 3FT molecule is inclined to the surface. Moreover, the SER spectrum of 2FT molecule predicts the existence of syn form of this molecule in the adsorbed state because of large enhancement of the carbonyl stretching mode. Concentration dependence of relative enhancement infers the monolayer formation on metal surface at a specific concentration, implying strong evidence of chemisorption of the adsorbates. The excitation profile of C identical withC symmetric stretching mode of 2FT suggests classical plasmon resonance as the main contributor to surface enhancement. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344530 TI - Preparation of Magnetic Inks: Adsorption of Macromolecular Dispersion Agents on Magnetic Metallic Iron Pigments and Electrical Charge Formation AB - The interaction between metallic iron pigments (MPs) and the actual constituents of magnetic ink is examined by first determining the adsorption isotherms and analyzing by FTIR the type of bonding that is established between the constituents and MP. Then, following the approach described in a previous paper using model compounds, the formation of electrical charges on MPs is examined through acoustophorometric (Electrokinetic amplitude or ESA) measurements. It is shown that the variation in ESA is related to charge transfer processes and reflects the MP-constituent interactions. Finally, adsorption competitions are shown and the importance of the method of preparation (order of mixing of the ingredients of the ink) is illustrated. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344531 TI - Characterization and CO2 Adsorptivity of Acid-Washed and Cation-Exchanged Natural Mordenites AB - A Japanese natural mordenite was modified by acid washing as well as cation exchange. Crystal structure, porosity, and active sites of the modified natural mordenites were characterized, and their CO2 adsorptivity were examined. Contraction as well as expansion of the mordenitic unit cell in the natural zeolite and distortion of zeolite structure by acid washing are plausibly shown by XRD. Long-term washing by nitric acid induces enlargement of micropore volume, increasing the micropore dimension but decreasing the large pore dimension over the mesoporous range. The degree of dehydration has an important role in CO2 adsorption. Metal ionic sites affect both the irreversible CO2 adsorption and the CO2 equilibrium adsorption at low concentration. Li+ ion-exchange brings about a great CO2 adsorption but reduces the CO2-surface interaction due to molecular size effect. Ca2+ ion-exchange has its main effect in the enhancement of CO2 desorption energy. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344532 TI - Estimation of Gelatin Layer Thickness on Polystyrene Particles by a Viscometric Study AB - A viscometric method has been used to determine the layer thickness of gelatin adsorbed on polystyrene (PS) particles. The layer thickness was found to be 28 nm regardless of the diameter of PS particles. The value of Einstein's coefficient, alpha0 for the relative viscosity-volume fraction relation was estimated to be 2.63. Since the PS latex particles were spherical, the value of alpha0 indicates that even at high shear rates, some aggregation still occurred in the dispersion. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344533 TI - The Effect of Head-Group on Selective Counterion Binding to Cationic Surfactants AB - Selectivity coefficients for the competitive adsorption of chloride, bromide, and iodide at the air/solution interface have been measured by a high accuracy ion flotation technique using the surfactant cations dodecylammonium, dodecylmethylammonium, dodecyldimethylammonium, and dodecyltrimethylammonium. Selective binding was found to depend on the nature of the head group, with selectivity increasing from primary to quaternary ammonium ions. Results are consistent with counterion binding arising from contact adsorption by mutual dehydration of head groups and counterions. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344534 TI - Cationic Surfactant Adsolubilization of 2-Naphthol and Naphthalene with Titanium Dioxide Having Dodecyl Chain AB - Adsolubilization of 2-naphthol and naphthalene by cationic surfactant-adsorbed layers formed on titanium dioxides with or without a dodecyl chain was investigated. The cationic surfactants used were dodecyltrimethylammonium bromide and 1,2-bis(dodecyldimethylammonio) ethane dibromide (2RenQ). It was found that the adsolubilized amounts of 2-naphthol and naphthalene increase and reach a maximum and then decrease for both surfactants and titanium dioxides with or without the dodecyl chain, where in the absence of surfactants the incorporated amount of naphthalene on the titanium dioxide with the dodecyl chain is markedly large. The adsolubilized amounts of 2-naphthol and naphthalene were enhanced with the titanium dioxide with the dodecyl chain by adsorption of surfactants, in particular 2RenQ. The admicellar partitioning coefficients also showed that naphthalene is adsolubilized preferentially rather than 2-naphthol and the surface treatment with the dodecyl chain enhances the adsolubilization for two adsolubilizates. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344535 TI - The Drosophila ovarian tumor gene is required for the organization of actin filaments during multiple stages in oogenesis. AB - The ovarian tumor gene is required during both early and late stages of oogenesis. Mutations produce a range of phenotypes, including agametic ovarioles, tumorous egg chambers, and late stage oogenic arrest. We demonstrate that each of these phenotypes is associated with specific aberrations in actin distribution. In the earliest case, ovarian tumor mutations cause actin filaments to accumulate ectopically in the fusome. This correlates with abnormal fusome morphology and arrested germ cell development in the germaria. Similarly, ovarian tumor function is required for the localization of actin that is essential for the maturation of ring canals. This defect gives rise to tumorous egg chambers in which germ cell numbers and morphology are profoundly aberrant. We also confirm that ovarian tumor is required for the formation of the nurse cell cytoplasmic actin array that is essential for the nonspecific transport of cytoplasmic contents to the oocyte during late oogenesis. Our data suggest that at this stage ovarian tumor controls the site where actin filaments initiate. Taken together, these studies suggest that the diverse ovarian tumor mutant phenotypes derive from the mislocalization of actin filaments, indicating a role for this gene in organizing the female germline cytoskeleton, and that the misregulation of actin can have profound effects on germ cell division and differentiation. PMID- 9344536 TI - Origin and differentiation of supernumerary midline glia in Drosophila embryos deficient for apoptosis. AB - Drosophila embryos deficient for programmed cell death produce 9 midline glia (MG) in addition to the wild-type complement of 3.2 MG/segment. More than 3 of the supernumerary MG derive from the MGP (MG posterior) lineage and the remainder from the MGA/MGM (MG anterior and middle) lineage. There is one unidentified additional neuron in the mesectoderm of embryos deficient for apoptosis. The supernumerary MG are not diverted from other lineages nor do they arise from an altered pattern of mitosis. Instead, these MG appear to arise from a normally existing pool of 12 precursor cells, larger than anticipated by earlier studies. During normal development, MG survival is dependent upon signaling to the Drosophila EGF receptor. The persistence of supernumerary MG in embryos deficient for apoptosis does not alter the spatial pattern of Drosophila EGF receptor signaling. The number and position of MG which express genes dependent upon EGF receptor function, such as pointed or argos, are indistinguishable from wild type. Genes of the spitz group are required for Drosophila EGF receptor function. Surviving MG in spitz group/H99 double mutants continue to express genes characteristic of the MG, but the cells fail to differentiate into ensheathing glia and are displaced from the nerve cord. PMID- 9344538 TI - The region encoded by the alternatively spliced exon IIIA in mesenchymal fibronectin appears essential for chondrogenesis at the level of cellular condensation. AB - Fibronectin in the extracellular matrix of tissues acts as a substrate for cell adhesion and migration during development. Heterogeneity in the structure of fibronectin is largely due to the alternative splicing of at least three exons (IIIB, IIIA, and V) during processing of a single primary transcript. Fibronectin mRNA alternative splicing patterns change from B+A+V+ to B+A-V+ during chondrogenesis. In this report, immunohistochemical analysis demonstrates that while fibronectin protein containing the region encoded by exon IIIB is present throughout the limb at all stages of development, fibronectin protein containing the region encoded by exon IIIA disappears from cartilaginous regions just after condensation in vivo and in high-density mesenchymal micromass cultures in vitro. Treatment of mesenchymal micromass cultures prior to condensation with an antibody specific for the region encoded by exon IIIA disrupts the formation of cellular condensations and inhibits subsequent chondrogenesis in a dose- and time dependent manner. Furthermore, microinjection of the exon IIIA antibody into embryonic chick limb primordia in vivo results in malformations characterized by smaller limbs and loss of limb skeletal elements. These results strongly suggest that the presence of the region encoded by exon IIIA in mesenchymal fibronectin is necessary for the condensation event that occurs during chondrogenesis. PMID- 9344537 TI - Expression of betacellulin and epiregulin genes in the mouse uterus temporally by the blastocyst solely at the site of its apposition is coincident with the "window" of implantation. AB - In the mouse, the process of implantation is initiated by the attachment reaction between the blastocyst trophectoderm and uterine luminal epithelium that occurs at 2200-2300 h on day 4 (day 1 = vaginal plug) of pregnancy. Several members of the EGF family are considered important in embryo-uterine interactions during implantation. This investigation demonstrates that the expression of two additions to the family, betacellulin and epiregulin, are exquisitely restricted to the mouse uterine luminal epithelium and underlying stroma adjacent to the implanting blastocyst. These genes are not expressed during progesterone maintained delayed implantation, but are rapidly switched on in the uterus surrounding the implanting blastocyst following termination of the delay by estrogen. These results provide evidence that expression of betacellulin and epiregulin in the uterus requires the presence of an active blastocyst and suggest an involvement of these growth factors in the process of implantation. PMID- 9344539 TI - Genes that induce apoptosis: transcriptional regulation in identified, doomed neurons of the Drosophila CNS. AB - Hormones and trophic factors provide cues that control neuronal death during development. These developmental cues in some way regulate activation of apoptosis, the mechanism by which most, if not all, developmentally programmed cell deaths occur. In Drosophila, apoptosis can be induced by the expression of the genes reaper, grim, or head involution defective. We demonstrate that prior to the death of a set of identifiable doomed neurons, these neurons accumulate transcripts of the reaper and grim genes, but do not accumulate transcripts of the head involution defective gene. Death of these doomed neurons can be suppressed by two manipulations: by increasing the levels of the steroid hormone 20-hydroxyecdysone or by decapitation. We have investigated the impact that these two manipulations have on reaper expression. Steroid treatment prevents the accumulation of reaper transcripts, whereas decapitation results in the accumulation of lower levels of reaper transcripts that are not sufficient to activate apoptosis. These data demonstrate that in vivo, reaper, and grim transcripts accumulate coordinately in a set of identified doomed neurons prior to the onset of apoptosis. These observations raise the possibility that products of the reaper and grim genes act in concert in postembryonic neurons to induce apoptosis. That reaper transcript accumulation is regulated by the steroid hormone titer and by the presence of the head is evidence that developmental factors control programmed cell death by regulating the expression of genes that induce apoptosis. PMID- 9344540 TI - Mesodermal guidance of pioneer axon growth. AB - Pioneer axons in insect legs are experimentally accessible model systems for the molecular identification and cellular localization of guidance cues regulating the path of axon growth. A detailed study of the Fe2 pioneer axons in the legs of the cockroach was performed to examine the diversity of guidance mechanisms. A detailed microscopic analysis of the axons at various points in their trajectory indicates that the Fe2 axons grow on a mesodermal substratum which contains the cues guiding their growth along a stereotyped path. An identified pair of muscle pioneer cells (MPC) are likely to play an important role in enabling the Fe2 growth cones to respond to mesodermal guidance cues. The addition of heparan sulfate, heparitinase, and phosphatidylinositol-specific phospholipase C to the medium perturbs the in situ path of growth of the Fe2 axons and the location of the MPC in cultured embryos. This indicates a role for heparan sulfate proteoglycans and glycosylphosphatidylinositol-anchored proteins in axon guidance. When these results are compared to those of similar experiments performed on the well-characterized Ti1 axons, they indicate significant differences in the mechanisms that are used for axon guidance. The Fe2 neurons are a good model for elucidating the mechanisms used to guide axon growth on nonmuscle mesodermal substrates often encountered in the periphery of vertebrate embryos. PMID- 9344541 TI - Serotonin plays an early role in the metamorphosis of the hydrozoan Phialidium gregarium. AB - Hydrozoan larvae normally metamorphose in response to an obligate external environmental cue. Application of certain artificial chemical stimuli will also induce metamorphosis. These chemicals and their inhibitors have been used to define and order some of the signal transduction events involved in this process. Results from this study show that exogenous application of serotonin (5-HT) will induce metamorphosis and that 5-HT immunoreactive cells are present in larvae when they are competent to metamorphose. The 5-HT inhibitors ketanserin, clozapine, and 5,7-DHT prevent metamorphosis from occurring as a response to a natural inducing stimulus. Additionally, 5-HT signaling occurs prior to both an influx of external Ca2+ from seawater and activation of protein kinase C, two other steps in the metamorphic signal transduction pathway. The neuropeptide LWamide, previously shown to induce metamorphosis in a related hydrozoan, Hydractinia echinata, also induced metamorphosis in Phialidium. When larvae were cotreated with LWamide and the 5-HT antagonist ketanserin, settlement occurred but was not followed by polyp morphogenesis. These results are used to present a model for the action of 5-HT during metamorphosis in Phialidium gregarium. PMID- 9344543 TI - Distinct neuronal lineages of the ascidian embryo revealed by expression of a sodium channel gene. AB - The ascidian larva contains tubular neural tissue, one of the prominent anatomical features of the chordates. The cell-cleavage pattern and cell maps of the nervous system have been described in the ascidian larva in great detail. Cell types in the neural tube, however, have not yet been defined due to the lack of a suitable molecular marker. In the present work, we identified neuronal cells in the caudal neural tube of the Halocynthia embryo by utilizing a voltage-gated Na+ channel gene, TuNa I, as a molecular marker. Microinjection of a lineage tracer revealed that TuNa I-positive neurons in the brain and in the trunk epidermis are derived from the a-line of the eight-cell embryo, which includes cell fates to epidermal and neural tissue. On the other hand, TuNa I-positive cells in the more caudal part of the neural tissue were not stained by microinjection into the a-line. These neurons are derived from the A-line, which contains fates of notochord and muscle, but not of epidermis. Electron microscopic observation confirmed that A-line-derived neurons consist of motor neurons innervating the dorsal and ventral muscle cells. Isolated A-line blastomeres have active membrane excitability distinct from those of the a-line derived neuronal cells after culture under cleavage arrest, suggesting that the A line gives rise to a neuronal cell distinct from that of the a-lineage. TuNa I expression in the a-line requires signals from another cell lineage, whereas that in the A-line occurs without tight cell contact. Thus, there are at least two distinct neuronal lineages with distinct cellular behaviors in the ascidian larva: the a-line gives rise to numerous neuronal cells, including sensory cells, controlled by a mechanism similar to vertebrate neural induction, whereas A-line cells give rise to motor neurons and ependymal cells in the caudal neural tube that develop in close association with the notochord or muscle lineage, but not with the epidermal lineage. PMID- 9344542 TI - A novel KH-domain protein mediates cell adhesion processes in Drosophila. AB - Adhesion of cells to one another and to extracellular matrices has major roles in morphogenetic processes during development. One important family of cell adhesion receptors are the integrins, which in Drosophila have crucial functions in at least two adhesion-mediated developmental events: embryonic muscle attachment and adhesion of the wing epithelia. We have cloned and characterized a gene (struthio) that is expressed in embryonic mesodermal and muscle cells, including cardioblasts, and epidermal muscle attachment sites in a pattern that is reminiscent of the expression pattern of the PS integrins. Maternal and zygotic transcripts are produced by this gene and encode similar proteins with two alternative carboxy tails. Both proteins contain identical KH domains, a protein sequence motif that is found in numerous proteins that interact with RNA. The struthio protein is highly homologous in a region including the KH domain to the mouse quaking and C. elegans gld-1 proteins, two developmentally important genes. Somatic homozygous clones of an embryonic lethal mutation in this gene (stru1A122) cause wing blisters and flight impairment, phenotypes which are associated with PS integrin subunit mutations. Thus, the struthio gene encodes a putative RNA-binding protein that appears to regulate some aspects of Drosophila integrin functioning. PMID- 9344544 TI - Rap1 overexpression reveals that activated RasD induces separable defects during Dictyostelium development. AB - One of the Dictyostelium ras genes, rasD, is expressed preferentially in prestalk cells at the slug stage of development and overexpression of this gene containing a G12T activating mutation causes the formation of aberrant multitipped aggregates that are blocked from further development (Reymond et al., 1986, Nature, 323, 340-343). The ability of the Dictyostelium rap1 gene to suppress this abnormal developmental phenotype was investigated. The rap1 gene and G12V activated and G10V negative mutant forms of the rap1 gene were independently linked to the rasD promoter and each construct used to transform M1, a Dictyostelium cell line expressing RasD[G12T]. Transformants of M1 that expressed Rap1 or Rap1[G12V] protein still formed multitipped aggregates, but most tips were able to complete development and form fruiting bodies. Cell lines showing this modified phenotype were designated ME (multitipped escape). The rap1[G10V] construct did not modify the M1 phenotype. These data suggest that overexpression of RasD[G12T] has two effects, the formation of a multitipped aggregate and a block in subsequent differentiation and that the expression of Rap1 or Rap1[G12V] reverses only the latter. Differentiation of ME cells in low density monolayers showed the identical low level of stalk and spore cell formation seen for M1 cells under the same conditions. Thus the cell autonomous defect in monolayer differentiation induced in the M1 strain was not corrected in the ME strain. Cell type-specific gene expression during the development of M1 cells is dramatically altered: prestalk cell-specific gene expression is greatly enhanced, whereas prespore-specific gene expression is almost suppressed (Louis et al., 1997, Mol. Biol. Cell, 8, 303-312). During the development of ME cells, ecmA mRNA levels were restored to those seen for Ax3, and tagB mRNA levels were also markedly reduced, although not to Ax3 levels. cotC expression in ME cells was enhanced severalfold relative to M1, although levels were still lower than those observed during the development of Ax3. The low expression of car1 mRNA during early development of the M1 strain remained low during the development of ME cells. These data are consistent with the idea that the expression of RasD[G12T] affects two independent and temporally separated events and that only the later defect is reversed by rap1. PMID- 9344545 TI - Conserved anterior boundaries of Hox gene expression in the central nervous system of the leech Helobdella. AB - Molecular developmental studies of fly and mouse embryos have shown that the identity of individual body segments is controlled by a suite of homeobox containing genes called the Hox cluster. To examine the conservation of this patterning mechanism in other segmented phyla, we here describe four Hox gene homologs isolated from glossiphoniid leeches of the genus Helobdella. Based on sequence similarity and phylogenetic analysis, the leech genes Lox7, Lox6, Lox20, and Lox5 are deemed to be orthologs of the Drosophila genes lab, Dfd, Scr, and Antp, respectively. Sequence similarities between Lox5 and Antp outside the homeodomain and phylogenetic reconstructions suggest that the Antennapedia family of Hox genes (as defined by Burglin, 1994) had already expanded to include at least two discrete Antp and Ubx/abdA precursors prior to the annelid/arthropod divergence. In situ hybridization reveals that the four Lox genes described in this study are all expressed at high levels within the segmented portion of the central nervous system (CNS), with variable levels of expression in the segmental mesoderm. Little or no expression was seen in peripheral ectoderm or endoderm, or in the unsegmented head region (prostomium). Each Lox gene has a distinct anterior expression boundary within one of the four rostral segments, and the anterior-posterior (AP) order of these expression boundaries is identical to that reported for the orthologous Hox gene products in fly and mouse. This finding supports the idea that the process of AP axis differentiation is conserved among the higher metazoan phyla with respect to the regional expression of individual Hox genes along that axis. One unusual feature of leech Hox genes is the observation that some genes are only expressed during later development -- beginning at the time of terminal cell differentiation -- whereas others begin expression at a much earlier stage, and their RNA ceases to be detectable shortly after the onset of expression of the 'late' Hox genes. The functional significance of this temporal disparity is unknown, but it is noteworthy that only the two 'early' Hox genes display high levels of mesodermal expression. PMID- 9344547 TI - A cellular stress model for the differential expression of glial lysosomal cathepsins in the aging nervous system. AB - Activation of the endosomal-lysosomal system and altered expression of various lysosomal hydrolases have been implicated in several senescence-dependent neurodegenerative disorders and occurs, to a lesser extent, in the course of normal brain aging. The progressive accumulation of autofluorescent, peroxidase positive astrocytic granules represents a highly consistent biomarker of aging in the vertebrate CNS. The sulfhydryl agent cysteamine greatly accelerates the accumulation of these glial inclusions in situ and in primary brain cell cultures. We previously determined that these glial inclusions are derived from abnormal mitochondria which undergo fusion with lysosomal elements in a complex autophagic process. In the present study, we demonstrate that cysteamine suppresses cathepsin B mRNA levels and immunoreactive protein in cultured astroglia, whereas cathepsin D mRNA and protein levels are significantly augmented by CSH exposure in these cells. Moreover, cathepsin D (but not cathepsin B) exhibits robust colocalization to the red autofluorescent inclusions. Concordant with our in vitro observations, cathepsin B immunoreactivity is prominent in the hypothalamic ventromedial nucleus which accumulates few autofluorescent glial inclusions during aging and is relatively inapparent in the heavily granulated hypothalamic arcuate nucleus. Conversely, cathepsin D is prominent in the aging arcuate nucleus where it colocalizes to the autofluorescent inclusions and exhibits scant immunoreactivity in the adjacent ventromedial nuclear complex. In senescent astroglia, oxidative stress may down regulate the cathepsin B gene as part of a concerted cellular stress (heat shock) response. Glial cathepsin D, on the other hand, resists stress-related inhibition and may play an important role in disposing of oxidatively modified mitochondria in the aging and degenerating nervous system. PMID- 9344546 TI - Equilin, a principal component of the estrogen replacement therapy premarin, increases the growth of cortical neurons via an NMDA receptor-dependent mechanism. AB - Regulation of both the outgrowth and the survival of neurons involved in cognitive function can have a significant impact on the function of neural networks involved in memory and other cognitive processes. Results of this investigation demonstrated that 17beta-estradiol and the estrogenic steroids estrone, estriol, mestranol, and equilin induced significant increases in cortical nerve cell growth. Of the neurotrophic estrogenic steroids, equilin was most efficacious. We therefore conducted an extensive analysis of equilin-induced neurotrophism. Equilin induced highly significant increases in the growth of both the macro and micro features of cortical nerve cell morphology. The growth promoting effects of equilin were present in both serum-containing and serum-free media, indicating that the growth-promoting effect of equilin is direct and not dependent upon factors present in serum. Analysis of the regional selectivity of equilin-induced neurotrophism in the cerebral cortex demonstrated that equilin significantly increased the growth of neurons from the frontal, temporal, and occipital regions, with neurons from the parietal region also influenced, though more modestly. We pursued the mechanism of equilin-induced neurotrophism and found that the growth-promoting effects of equilin were completely abolished in the presence of the glutamatergic NMDAreceptor antagonist AP5. Equilin is a major component of Premarin, the leading prescribed pharmaceutical for estrogen replacement therapy for postmenopausal women in the United States. Results of this investigation have the potential of influencing the application and design of therapeutic agents for the prevention of cognitive decline in estrogen deficient women and for the prevention of Alzheimer's disease in postmenopausal women, a group that comprises a large sector of the population, the size of which will continue to grow in the coming decades. PMID- 9344548 TI - Increased expression of cathepsin D in retrosplenial cortex of MK-801-treated rats. AB - Single administration of a high dose of an uncompetitive NMDA receptor antagonist dizocilpine maleate (MK-801)-results in transient neuronal vacuolization and cell death in retrosplenial cortex in rodents. In this study expression of cathepsin D (CatD), a major lysosomal aspartic protease, was investigated in brains of female rats treated with 1, 5, or 10 mg/kg of MK-801. Northern blot analysis demonstrated that the CatD mRNA level was moderately increased in retrosplenial cortex 24 h-7 days after the treatment. Concomitantly, increased CatD immunoreactivity was observed, predominantly in the degenerating neurons in layer III of retrosplenial cortex. Neuronal response was spatially distinguished from glial reactivation marked by increased mRNA and protein levels of glial fibrillary acidic protein, as demonstrated by Northern blot and immunohistochemistry in retrosplenial cortex 24 h-7 days after MK-801 treatment. These data suggest that activation of the lysosomal proteolytic system of neurons may play a role in MK-801-evoked neurodegeneration. PMID- 9344549 TI - In vitro phosphorylation of cytoskeletal proteins in the rat cerebral cortex is decreased by propionic acid. AB - In the present study we demonstrate that propionic acid (PA), a metabolite that accumulates in large amounts in propionic acidemia, is able to decrease in vitro incorporation of [32P]ATP into neurofilament subunits (NF-M and NF-L) and alpha- and beta-tubulin. Considering that the endogenous phosphorylating system associated with the cytoskeletal fraction contains cAMP-dependent protein kinase (PKA), Ca2+/calmodulin protein kinase II (CaMKII), and protein phosphatase 1 (PP1), we first assayed the effect of the acid on the kinase activities by using the specific activators cAMP and Ca2+/calmodulin or the inhibitors PKAI or KN-93 for PKA and CaMKII, respectively. Results demonstrated that the acid totally inhibited the stimulatory effect of cAMP and interfered with the inhibitory effect of PKAI. In addition, PA partially prevented the stimulatory effect of Ca2+/calmodulin and interfered with the effect of KN-93. In addition, we demonstrated that PA totally inhibited in vitro dephosphorylation of neurofilament subunits and tubulins mediated by PP1 in brain slices pretreated with the acid. Taken together, these results demonstrate that PA inhibits the in vitro activities of PKA, CaMKII, and PP1 associated with the cytoskeletal fraction of the cerebral cortex of rats. This study suggests that PA at the same concentrations found in tissues from propionic acidemic children may alter phosphorylation of cytoskeletal proteins, which may contribute to the neurological dysfunction characteristic of propionic acidemia. PMID- 9344550 TI - Transient damage to the axonal transport system without Wallerian degeneration by acute nerve compression. AB - The aim of this study was to examine whether acute nerve compression damages an axonal transport system based on microtubules and how the fibers recover after the compression. A 5-mm segment of the tibial nerve of male wistar rat was compressed with a specially designed clip. Functional recovery was assessed using Tibial Nerve Functional Index (TFI). Rats were sacrificed each day from Day 0 to Day 2 and every 2 days between Day 4 and Day 10. For immunohistochemical analysis of the tibial nerve, the proximal uncompressed, the middle compressed, and the distal uncompressed segments of each section were assessed under immunofluoroscent microscopy for anti-dynein, anti-tubulin, and anti neurofilament antibodies staining. In rats whose tibial nerve was compressed by 25 g/mm2 of pressure for 5 min, staining of dynein and mirotubules in the compressed portion were obscure on Days 4-8, suggesting that the microtubules based axonal transport system was temporarily damaged, while neurofilaments were retained. In contrast, in the distal portion, anti-neurofilament staining showed no abnormality throughout the experimental period, indicating that Wallerian degeneration did not occur. We conclude that acute nerve compression can cause transient damage to the axonal transport system in nerve fibers without Wallerian degeneration. PMID- 9344551 TI - New growth of axons in the cochlear nucleus of adult chinchillas after acoustic trauma. AB - This study determined the effect of acoustic overstimulation of the adult cochlea on axons in the cochlear nucleus. Chinchillas were exposed to an octave-band noise centered at 4 kHz at 108 dB sound pressure level for 1.75 h. One chinchilla was never exposed to the noise, and several others had one ear protected by an ear plug or prior removal of the malleus and incus. Exposure of unprotected ears caused loss of inner and outer hair cells and myelinated nerve fibers, mostly in the basal half of the cochlea. Cochlear nerve fiber degeneration, ipsilateral to the exposed ears, was traced to regions of the cochlear nucleus representing the damaged parts of the cochlea. In silver impregnations of a deafferented zone in the posteroventral cochlear nucleus, the concentration of axons decreased by 43% after 1 month and by 54% after 2 months. However, by 8 months, the concentration of thinner axons, with diameters of less than 0.46 microm, increased by 46-90% over that at 2 months. The concentration of axons with larger diameters did not change. Between 2 and 8 months small axonal endings appeared next to neuronal cell bodies. This later increase of thinner axons and endings is consistent with a reactive growth of new axons of relatively small diameter. The emergence of small perisomatic boutons suggests that the new axons formed synaptic endings, which might contribute to an abnormal reorganization of the central auditory system and to the pathological changes that accompany acoustic overstimulation. PMID- 9344552 TI - A nonimmunosuppressant FKBP-12 ligand increases nerve regeneration. AB - The immunosuppressant drugs FK506 and cyclosporin A inhibit T-cell proliferation via a common mechanism: calcineurin inhibition following binding to their respective binding proteins, the peptidyl prolyl isomerases FKBP-12 and cyclophilin A. In contrast, FK506, but not cyclosporin A, accelerates nerve regeneration. In the present study, we show that the potent FKBP-12 inhibitor V 10,367, which lacks the structural components of FK506 required for calcineurin inhibition, increases neurite outgrowth in SH-SY5Y neuroblastoma cells and speeds nerve regeneration in the rat sciatic nerve crush model. In SH-SY5Y cells, V 10,367 increased the lengths of neurite processes in a concentration-dependent (between 1 and 10 nM) fashion over time (up to 168 h). Daily subcutaneous injections of V-10,367 accelerated the onset of clinical signs of functional recovery in the hind feet compared to vehicle-treated control animals. Interdigit distances (between the first and fifth digits) measured on foot prints obtained during walking showed an increase in toe spread in V-10,367-treated rats compared to vehicle-treated controls. Electron microscopy demonstrated larger regenerating axons distal to the crush site in the sciatic nerve from V-10,367-treated rats. Quantitation of axonal areas in the soleus nerve revealed a shift to larger axonal calibers in V-10,367-treated rats (400 or 200 mg/kg/day); mean axonal areas were increased by 52 and 59%, respectively, compared to vehicle-treated controls. FKBP-12 ligands lacking calcineurin inhibitory activity represent a new class of potential drugs for the treatment of human peripheral nerve disorders. PMID- 9344553 TI - Chiasmatic specificity in the regenerating mammalian optic nerve. AB - The mammalian central nervous system is capable of regenerating; however, there is no evidence that the regenerating axons can navigate along their normal pathways and reestablish topographically organized projections: essential for functional return of vision. Here retinal ganglion cells in the opossum Monodelphis were birthdated with tritiated thymidine on the sixth postnatal day (P6), before being lesioned in the temporal retina at P8. Retrograde tracing with horseradish peroxidase injected into the ipsilateral optic tract at P24 showed that the temporal crescent had reformed behind the retinal lesion. By comparisons of cell and thymidine counts from lesioned and control regions of retina, it was estimated that about 40% of the normal number of ganglion cells are able to regenerate into the ipsilateral optic tract following a lesion in the temporal retina at P8. A clear line of decussation (separation of ipsilateral and contralateral projections) reformed in the lesioned temporal retina and regenerating ganglion cells labeled with DiI were turned at appropriate points on passing through the optic chiasm. This is evidence of chiasmatic specificity with regard to lesioned retinal ganglion cells regenerating into the ipsilateral optic tract. PMID- 9344554 TI - Differential spine loss and regrowth of striatal neurons following multiple forms of deafferentation: a Golgi study. AB - Golgi-Cox method and morphometric analyses were used to study the plasticity of striatal medium spiny I neurons in 6-month-old C57BL/6N mice after unilateral or bilateral lesion of the cerebral cortex or combined lesions of the ipsilateral cerebral cortex and intralaminar thalamus. In adult mouse, unilateral lesions of the cerebral cortex did not result in a net gain or loss of linear dendritic length in a randomly selected population of striatal medium spiny I neurons. In addition, there was a well-defined time course of striatal spine loss and replacement occurring after a unilateral cortical lesion. By day 3 postlesion the average 20-microm dendritic segment had lost 30% of the unlesioned control spine value, reached its nadir, lost 45.5%, at 10 days postlesion, and recovered to 80% of unlesioned control levels by 20 days postlesion. The recovery of spines was blocked by a secondary lesion on the contralateral cortex but not on the ipsilateral intralaminar thalamus. These data suggest that striatal medium spiny I neurons of adult mice have a remarkable capacity for plasticity and reactive synaptogenesis following a decortication. The recovery of spine density is primarily induced by axonal sprouting of survival homologous afferent fibers from the contralateral cortex. PMID- 9344555 TI - Morphological plasticity induced in the phrenic nucleus following cervical cold block of descending respiratory drive. AB - Morphological plasticity occurs in the phrenic nucleus within hours following an ipsilateral C2 spinal cord hemisection. The plasticity has been associated with the unmasking of a latent respiratory pathway (the crossed phrenic pathway) which allows recovery of the hemidiaphragm paralyzed by the hemisection during a reflex known as the crossed phrenic phenomenon. This study tests if the plasticity is induced by the generalized effects of spinal cord trauma or the more specific effect of interrupting the main descending respiratory drive to phrenic motoneurons. Electron microscopic quantitative morphometric analysis of the phrenic nucleus neuropil was carried out on four Sprague-Dawley rats (200-250 g) sacrificed 4 h following unilateral reversible cold block of the descending bulbospinal respiratory drive at the second cervical segment of the spinal cord (C2). The data from four sham-operated control animals were compared with those of the experimental group. The following morphological alterations were documented in cold block animals compared to controls: (1) a significant increase in the number of multiple synapses (i.e., terminals with synaptic active zones contacting two or more postsynaptic profiles in the same plane of section), (2) a significant increase in the number of dendrodendritic appositions, and (3) a significant increase in the length of symmetric and asymmetric synaptic active zones. The above changes are similar to the changes induced in the phrenic nucleus following C2 hemisection. We conclude therefore, that injury to the spinal cord is not a requirement for this type of morphological plasticity in the phrenic nucleus, but rather the induced changes are activity-dependent and are likely caused by the interruption of the descending bulbospinal respiratory drive to the phrenic nucleus. PMID- 9344556 TI - GABA and glutamate levels in the substantia nigra reticulata following repetitive cerebral ischemia in gerbils. AB - Repetitive cerebral ischemia produces more severe damage than a similar single duration insult. We have previously shown that, in gerbils, damage in the substantia nigra reticulata (SNr) is seen with repetitive insults rather than a single insult. We have also shown that there is a progressive decrease in the extracellular GABA in the striatum in the days preceding such damage, speculating that a loss of GABA may be in part responsible for this damage. This study evaluates the GABA levels in the SNr in animals exposed to repetitive ischemic insults. Each animal received a total of three ischemic insults of 3-min duration at hourly intervals. In vivo microdialysis was carried out to analyze the GABA and glutamate dialysate levels on Days 1, 3, 5, 7, and 14 following the ischemic insult. In the control and treated (ischemic) animals, there was a significant increase in the GABA levels with the introduction of nipecotic acid on Days 1, 3, 5, and 14. However, on Day 7 there was a significant attenuation in the GABA response to nipecotic acid in the treated animals in comparison to the controls. The glutamate levels in the treated animals were similar to the control animals on Days 1, 3, 5, and 7. However, on Day 14 the glutamate levels were significantly lower than on previous days. Our experiments for the first time measure extracellular glutamate and GABA responses in the SNr in animals exposed to repetitive ischemic insults. Our experiments show that there is a significant decrease in the GABA concentrations at a time when ischemic damage is developing in this region. This confirms our hypothesis that a decrease in GABA may be one factor contributing to neuronal damage during the period following repetitive ischemic insults. Further, the rebound increase in GABA levels on Day 14 with a concomitant fall in glutamate levels would indicate that reparative processes are still active in the 2 weeks following the insult. PMID- 9344557 TI - Expression of Fos, Jun, and Krox family proteins in Alzheimer's disease. AB - Apoptosis is an active process of cell death characterized by distinct morphological features and is often the end result of a genetic program of events, i.e., programmed cell death (PCD). There is growing evidence supporting a role for apoptosis and/or PCD in Alzheimer's disease (AD), based on DNA fragmentation studies and recent findings of increased levels of inducible transcription factors (ITFs) such as c-Jun in AD brains. We have characterized the expression of a large range of ITFs (c-Fos, Fos B, Fos-related antigens, c Jun, Jun B, Jun D, Krox20, and Krox24) using multiple antisera in AD postmortem hippocampi and compared this with human control hippocampi as well as Huntington's disease hippocampi and human epilepsy biopsy tissue. We found little evidence of nuclear expression of any ITF except c-Jun in the human postmortem tissue, compared with nuclear staining in biopsy tissue. We found some evidence for increased levels of c-Jun and Krox24 protein and krox24 mRNA in the CA1 region of AD hippocampi, suggesting that PCD may be involved in the pathogenesis of AD. In general, staining characteristics of ITFs varied with different antisera directed against the same protein, indicating the need for caution when interpreting results. PMID- 9344558 TI - Immunohistochemical study of GABA(A) receptor beta2/3 subunits in the hippocampal formation of aged brains with Alzheimer-related neuropathologic changes. AB - In AD, it is hypothesized that one factor contributing to the vulnerability of neurons is a delicate balance of excitatory and inhibitory inputs. To examine this hypothesis we have initiated a number of studies examining the role of the excitatory neurotransmitter glutamate and the inhibitory neurotransmitter gamma aminobutyric acid (GABA) in the neurodegeneration of AD. As an initial investigation into the GABAergic system in AD, we employed immunocytochemical techniques and examined the distribution and density of the GABAA receptor subunits beta2/3 within the hippocampus of 13 subjects with a clinical diagnosis of AD and 6 nondemented elderly subjects. Collectively, these 19 subjects presented with a broad range of pathologic severity (i.e., Braak stages I-VI). Density measurements of nine hippocampal regions demonstrated highest levels of beta2/3 immunolabeling in the inner molecular layer of the dentate gyrus > CA1 > CA2, while the lowest levels were found in the granular layer of the dentate gyrus < or = CA4 < CA3 field. Despite these regional variations no significant difference in the mean density of beta2/3 immunolabeling was observed when comparing the pathologically mild (stages I and II), moderate (stages III and IV), and severe (stages V and VI) groups. These data suggest that in the hippocampus receptor subunits associated with GABAergic neurotransmission are relatively maintained even until the terminal stages of the disease. PMID- 9344559 TI - High dose baclofen is neuroprotective but also causes intracerebral hemorrhage: a quantal bioassay study using the intraluminal suture occlusion method. AB - Agonists of the GABA-A receptor are neuroprotective after experimental stroke, but studies of GABA-B agonists have contradicted each other. To further investigate whether GABA-B agonists may be neuroprotective, we devised a quantal bioassay using the intraluminal occlusion method of inducing reversible cerebral ischemia. Subjects underwent middle cerebral artery occlusion for varying amounts of time, ranging from 5 to 90 min. Behavioral outcome was measured 48 h later with a quantal observational scale: score of abnormal given for any one of asymmetric forepaw flexion on tail lift, asymmetric grip, circling, reduced exploration, seizures, or death. To the grouped response data the logistic equation was used to find the ED50, the duration of occlusion that caused one half of the subjects to be abnormal. To find the potency ratio for each drug, we divided the ED50 for treatment by that for vehicle. We administered baclofen, a GABA-B agonist, intraperitoneally 5 min after the onset ofischemia. Baclofen (20 mg/kg) was neuroprotective (potency ratio of 3.0, P < 0.05), but a lower dose (10 mg/kg) was not. However, both doses of baclofen caused significantly more intracerebral hemorrhages than control. In awake animals, both baclofen doses caused significant increases in mean arterial pressure, but no changes in other cardiorespiratory variables. The glutamate antagonist MK-801, the GABA-A agonist muscimol, and hypothermia were all protective using the bioassay (potency ratios ranging from 1.5 to 3.0). We conclude that although baclofen (20 mg/kg) may be neuroprotective, its utility is complicated by postischemic hypertension and cerebral hemorrhages. PMID- 9344560 TI - Influence of ischemia and reperfusion on the course of brain tissue swelling and blood-brain barrier permeability in a rodent model of transient focal cerebral ischemia. AB - Brain swelling is a serious complication associated with focal ischemia in stroke and severe head injury. Experimentally, reperfusion following focal cerebral ischemia exacerbates the level of brain swelling. In this study, the permeability of the blood-brain barrier has been investigated as a possible cause of reperfusion-related acute brain swelling. Blood-brain barrier disruption was investigated using Evans Blue dye and [14C]aminoisobutyric acid autoradiography in a rodent model of reversible middle cerebral artery (MCA) occlusion. Acute brain swelling and cerebral blood flow (CBF) during ischemia and reperfusion were analyzed from double-label CBF autoradiograms after application of the potent vasoconstrictor peptide endothelin-1 to the MCA. Ischemia was apparent within ipsilateral MCA territory, 5 min after endothelin-1 application to the exposed artery. Reperfusion, examined at 30 min and 1, 2, and 4 h, was gradual but incomplete within this time frame in the core of middle cerebral artery territory and associated with significant brain swelling. Ipsilateral hemispheric swelling increased over time to a maximum (>5%) at 1-2 h after endothelin-1 but was not associated with a significant increase in the ipsilateral transfer constant for [14C]aminoisobutyric acid over this time frame. These results indicate that endothelin-1 induced focal cerebral ischemia is associated with an acute but reversible hemispheric swelling during the early phase of reperfusion which is not associated with a disruption of the blood-brain barrier. PMID- 9344561 TI - The distribution of beta-amyloid precursor protein in rat cortex after systemic kainate-induced seizures. AB - In the current study we employed immunohistochemical techniques to identify neuronal and glial cells in specific brain areas that modulate beta-amyloid precursor protein (betaAPP) synthesis following kainate-induced seizures. In addition, antibodies directed against the FOS protein, which is generated by activation of the immediate early gene c-fos and is temporally associated with ongoing seizure activity, were used to identify transneuronal pathways activated after kainate-induced seizures (KIS). It was therefore possible to correlate the appearance of activated neuronal pathways identified by FOS-like immunoreactivity (LI) and PAPP-LI in alternate sections. In addition, we employed immunohistochemical procedures to characterize morphological changes in neuronal and glial cells following kainate-induced seizures in both young and adult rats. Our results demonstrate a specific pattern of FOS-LI induced by kainate injection. In older animals FOS-LI spreads out from limbic cortical regions, including the piriform and entorhinal cortex, to other cortical regions, including the parietal and somatosensory cortices. Seizures were associated with decrease in neuronal betaAPP-LI in both young and adult rats, whereas glial betaAPP-LI markedly increased. The increase in betaAPP-LI glia was far more extensive in adult than in young rats and the anatomical distribution of betaAPP LI glia was grossly correlated with FOS-LI. The spread of betaAPP-LI follows seizure-activated transsynaptic pathways. It is likely that the sequence of events following kainate injection is initially triggered by c-fos gene expression, which is rapidly followed by modulation of betaAPP synthesis in parallel to, or preceding, morphological changes of both microglia and astrocytes. The present study, which extensively characterized early changes in c fos expression and betaAPP-LI in glia following kainate-induced seizures, is a potentially useful animal model for the in vivo study of numerous facets of betaAPP synthesis and the possible role of such processes in Alzheimer's disease. PMID- 9344562 TI - Ultrastructural changes in the deep cortical pyramidal cells of infant rats with inherited hydrocephalus and the effect of shunt treatment. AB - Pathological changes in the cortical gray matter in infantile hydrocephalus vary with the age at onset and may not be reversible with shunt treatment. We have used electron microscopy to investigate the sequence of pathological change and the effect of shunt treatment on layer VI pyramidal cells from infant H-Tx rats with inherited early-onset hydrocephalus. Tissue was prepared from the frontal and visual cortex of control and hydrocephalic rats at 4, 11, and 21 days after birth, together with 21-day rats previously treated with ventriculosubcutaneous shunts at 4-5 or 10-11 days after birth. Both cortical regions gave similar results but the effects were more severe in the visual cortex. In the early stages of hydrocephalus, the pyramidal cells were in clusters with fewer mature dendrites and less cytoplasmic organization than those in control rats, and some neuronal processes were vacuolated. In intermediate hydrocephalus the changes were more severe, with vacuolated cytoplasm, fewer cytoplasmic organelles, frequent swollen processes, and infrequent synapses. In advanced hydrocephalus at 21 days, many neurons showed degenerative changes, with edematous Golgi and dilated endoplasmic reticulum, distorted mitochondria, and single ribosomes. The neuropil contained many spongy areas with distended profiles. Shunt treatment prevented most of the changes if carried out at 4 days. Shunt treatment at 11 days also gave a dramatic recovery at the cellular level, but there were more immature pyramidal cells and edematous processes in the neuropil than in the 4 day-treated rats. The changes in hydrocephalus are consistent with progressive neuronal damage, which is largely prevented by early shunt treatment. PMID- 9344563 TI - Plasma protein extravasation induced in the rat dura mater by stimulation of the parasympathetic sphenopalatine ganglion. AB - It has been proposed that migraine could result from a neurogenic inflammation of the dura mater. According to this theory, inflammation could be initiated by an axon reflex of nociceptive nerve fibers, but the trigger of this axon reflex remains poorly understood. Previous works have shown that parasympathetic agonists can activate mast cells and/or sensory C-fibers, inducing pain and inflammation. The aim of the present work was to determine whether the activation of intracranial parasympathetic nerve fibers could trigger an inflammatory mechanism within the rat dura mater. Activation of the intracranial parasympathetic system was achieved by electrical stimulation of the sphenopalatine ganglion (SPG). The development of a neurogenic inflammation was estimated either by microscopic examination or by quantitative measurement of plasma protein extravasation (PPE) in the dura. To determine the respective roles of the parasympathetic and sensory innervations, two groups of rats were pretreated either with atropine or with capsaicin. Stimulation of the SPG induced a PPE increase of about 200% in the stimulated side on the dura mater. Extravasated material was mainly concentrated around small blood vessels. This extravasation was significantly reduced by capsaicin pretreatment and completely abolished by atropine. Infusion of carbachol in the common carotid artery induced PPE in the ipsilateral dura comparable to that induced by electrical stimulation of the SPG. This extravasation was also blocked by atropine infusion. These data indicate for the first time that the parasympathetic nervous system can trigger a neurogenic inflammation in the dura via muscarinic cholinergic receptors. Sensory C-fibers seem to play a role in this phenomenon. With respect to the potential autonomic imbalance described in the etiology of various types of vascular headaches, such a mechanism could be important in inducing attacks. PMID- 9344564 TI - Ultrastructural changes of sympathetic neurons following neurotrophin 3 antiserum treatment in young rat. AB - We have previously demonstrated that neurotrophin-3 antiserum administration to rats during the first 2 postnatal weeks results in a massive reduction of neurons in the superior cervical ganglion. In the present study, an ultrastructural analysis was undertaken to elucidate the mechanism by which neurotrophin-3 deprivation causes neuronal death. Newborn and 4-week-old rats were injected with either neurotrophin-3 antiserum or normal rabbit serum or used without injection. Superior cervical ganglia from each animal were examined by routine electron microscopy. Most neurons in the ganglia from untreated rats had a large and round nucleus with one or two nucleoli. Chromatin within the nucleus was evenly distributed. A double-layer nuclear membrane could be distinguished and the cytoplasm contained abundant organelles. Treatment with neurotrophin-3 antiserum for 24 h in neonates resulted in chromatin clumping in the nucleus of many neurons. The nuclear membrane became rough and occasionally folded. In the cytoplasm, the Golgi apparatus was disrupted. Three days after treatment, these changes became more obvious. The chromatin in the nucleus was often aggregated and marginalized. Vacuolation was present in many membranous organelles throughout the cytoplasm. Although neurotrophin-3 antiserum given to 4-week-old rats had little effect on overall neuronal numbers (Tafreshi, Zhou, and Rush, unpublished), a few neurons, undergoing either apoptotic or cytolytic cell death, were identified 7 days later. Most affected neurons were located near small blood vessels or capillaries and were associated with numerous nonneuronal cells. The debris of degenerating neurons were surrounded by the processes of glia cells. These findings support the view that loss of endogenous neurotrophin-3 following neutralization with specific antibody leads to activation of apoptotic pathways within the affected neurons. However, the presence of neurons dying as a result of cytolysis suggests that other mechanisms may also be involved. PMID- 9344565 TI - Quantitative analysis of microglial reaction to a cortical excitotoxic lesion in the early postnatal brain. AB - This study was designed to quantify the microglial response following an injection of N-methyl-D-aspartate (NMDA) into the sensorimotor cortex of 6-day old rats. After survival times ranging from 10 h to 28 days, cryostat sections were processed for the demonstration of microglial cells by means of tomato lectin histochemistry. The injection of NMDA caused an extensive primary lesion involving the neocortex, the rostral hippocampus, and rostral thalamus. In addition, secondary retrograde/anterograde degeneration was also observed in the ventrobasal (VB) complex of the thalamus. Microglial reactivity was already present at 10 h postlesion and restricted to areas of neuronal degeneration. Quantitative analysis was performed on digitized images using NIH Image software and a Macintosh computer. The method is based on densitometric ratios, referred to as the "reactivity grade," between the ipsilateral lesion side and the contralateral control side. Measurements were made to determine a possible increase in the number of microglial cells as well as an increase in lectin binding. The analysis showed that microglial reactivity in areas of primary degeneration peaked at 3 days postlesion, when it was significantly (P < 0.01) higher in comparison to saline-injected litter mates. Microglial response in the cerebral neocortex, showing the highest reactivity grade, as well as in other areas of primary degeneration, returned to control levels by Day 7. Microglial response in the VB complex also peaked at Day 3 (P < 0.05) but maintained this level of reactivity until 7 days postlesion (P < 0.01). PMID- 9344566 TI - Insulin-like growth factor-1 (IGF-1) improves both neurological motor and cognitive outcome following experimental brain injury. AB - We evaluated the efficacy of insulin-like growth factor-1 (IGF-1) in attenuating neurobehavioral deficits following lateral fluid percussion (FP) brain injury. Male Sprague-Dawley rats (345-425 g, n = 88) were anesthetized and subjected to FP brain injury of moderate severity (2.4-2.9 atm). In Study 1, IGF-1 (1.0 mg/kg, n = 9) or vehicle (n = 14) was administered by subcutaneous injection at 15 min postinjury and similarly at 12-h intervals for 14 days. In animals evaluated daily for 14 days, IGF-1 treatment attenuated motor dysfunction over the 2-week period (P < 0.02). In Study 2, IGF-1 (4 mg/kg/day, n = 8 uninjured, n = 13 injured) or vehicle (n = 8 uninjured, n = 13 injured) was administered for 2 weeks via a subcutaneous pump implanted 15 min postinjury. IGF-1 administration was associated with increased body weight and mild, transient hypoglycemia which was more pronounced in brain-injured animals. At 2 weeks postinjury (P < 0.05), but not at 48 h or 1 week, brain-injured animals receiving IGF-1 showed improved neuromotor function compared with those receiving vehicle. IGF-1 administration also enhanced learning ability (P < 0.03) and memory retention (P < 0.01) in brain-injured animals at 2 weeks postinjury. Taken together, these data suggest that chronic, posttraumatic administration of the trophic factor IGF-1 may be efficacious in ameliorating neurobehavioral dysfunction associated with traumatic brain injury. PMID- 9344567 TI - Chronic spinal nerve ligation induces changes in response characteristics of nociceptive spinal dorsal horn neurons and in their descending regulation originating in the periaqueductal gray in the rat. AB - We studied whether a chronic neuropathy induced by unilateral spinal nerve ligation changes the response characteristics of spinal dorsal horn wide-dynamic range (WDR) neurons or their periaqueductal gray (PAG)-induced descending modulation. Experiments were performed in rats with behaviorally demonstrated allodynia induced by spinal nerve ligation and in a group of nonneuropathic control rats. The stimulus-response functions of WDR neurons for mechanical and thermal stimuli and the modulation of their peripherally evoked responses by electrical stimulation of the PAG were determined under pentobarbital anesthesia. The results showed that neuropathy caused a significant leftward shift in stimulus-response functions for mechanical stimuli. In contrast, stimulus response functions for noxious heat stimuli in the neuropathic limb were, if anything, shifted rightward, although this shift was short of statistical significance. In neuropathic rats, PAG stimulation produced a significantly stronger attenuation of spinal neuronal responses induced by noxious heat in the unoperated than in the operated side. At the intensity that produced attenuation of noxious heat stimuli, PAG stimulation did not produce any significant change in spinal neuronal responses evoked by mechanical stimuli either from the operated or the nonoperated hindlimb of the neuropathic rats. Spontaneous activity of WDR neurons was higher in the operated side of neuropathic rats than in control rats. Afterdischarges evoked by peripheral stimuli were observed in 1/16 of the WDR neurons ipsilateral to spinal nerve ligation and not at all in other experimental groups. The WDR neurons studied were not activated by innocuous or noxious cold stimuli. The results indicate that spinal nerve ligation induces increased spontaneous activity and enhanced responses to mechanical stimuli in the spinal dorsal horn WDR neurons, whereas noxious heat evoked responses are not significantly changed or if anything, attenuated. Moreover, the inhibition of noxious heat stimuli by PAG stimulation is attenuated in the neuropathic side. It is proposed that the observed changes in the response characteristics of the spinal dorsal horn WDR neurons and in their descending modulation may contribute to the neuropathic symptoms in these animals. PMID- 9344568 TI - Critical periods of basic fibroblast growth factor and brain-derived neurotrophic factor in the development of the chicken cochleovestibular ganglion in vitro. AB - The temporal roles of brain-derived neurotrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) in the development of sensory neurons have been studied in a cell culture preparation which models normal embryonic inner ear development (normocytic). Previous studies showed that FGF-2 stimulated migration and differentiation of ganglion cells for the first 2 days in vitro, but after 5 days led to degeneration, implicating other factors in their later development. To see if BDNF could be such a factor, otocysts were explanted from white leghorn embryos at the time when ganglion cell precursors normally start migrating from the otic epithelium. Cultures were grown in a defined medium, either with or without human recombinant FGF-2 for 2 days or with BDNF. On Day 3, FGF-2 was replaced either with BDNF in defined medium or with defined medium only. Measurements of neuroblast migration and neurite outgrowth were made by time lapse imaging in living cultures. In cultures receiving BDNF on Day 3, cell migration and neurite outgrowth from the explant increased for more than 3 weeks but not in cultures receiving only defined medium from Day 3. Cultures did not survive more than 3-4 days when receiving either BDNF in defined medium or defined medium alone from the first day. A neutralizing antibody to BDNF inhibited neuronal migration and neurite outgrowth, and it also blocked the effects of exogenous BDNF. BDNF did not enhance the effects of FGF-2 by interacting with it. These experiments defined a temporal sequence in which FGF-2 acts early in development, while BDNF affects a later stage. PMID- 9344569 TI - Sensory and motor denervation influence epidermal thickness in rat foot glabrous skin. AB - Denervation in man often results in shiny, dry, thin skin. A previous study has shown that the epidermis of glabrous skin in the rat becomes approximately 40% thinner within 1 week following sciatic nerve transection, but which nerve fiber type or types influence epidermal thickness is unknown. In this study, we compared the effects on the epidermis of selective sensory, motor, and sympathetic denervation. Protein gene product 9.5 and calcitonin gene-related peptide immunocytochemical staining were used to determine the extent of denervation of epidermis, dermis, and sweat glands in the footpads. Epidermal thickness of the glabrous plantar skin of the foot was measured. To verify the specificity and reliability of each animal model, the relevant regions of the peripheral nervous system were examined by light or electron microscopy or both. Epidermal thickness decreased significantly following sciatic nerve transection (58% of control, P < 0.05) and dorsal root ganglionectomy (59%; P < 0.05). The thickness also decreased following lumbar ventral rhizotomy (61%; P < 0.01), destruction of lumbar spinal motor neurons (66%; P < 0.05), and botulinum toxin induced paralysis of the tibialis anterior and gastrocnemius muscles (70%; P < 0.05). A slight decrease followed dorsal rhizotomy (84%; P < 0.01). In contrast, no significant alterations in epidermal thickness were detected following sham operation and sympathectomy. Epidermal thinning was paralleled by reductions in the amounts of transcripts for glyceraldehyde-3-phosphate dehydrogenase and beta actin. These results suggest that selective loss of both sensory and motor fibers to the hind limb can contribute to reducing epidermal thickness in rat foot glabrous skin. PMID- 9344570 TI - Spinal cord injury and anti-NGF treatment results in changes in CGRP density and distribution in the dorsal horn in the rat. AB - Spinal cord injury (SCI) results in chronic pain states in which the underlying mechanism is poorly understood. To begin to explore possible mechanisms, calcitonin gene-related peptide (CGRP), a neuropeptide confined to fine primary afferent terminals in laminae I and II in the dorsal horn of the spinal cord and implicated in pain transmission, was selected. Immunocytochemical techniques were used to examine the temporal and spatial distribution of CGRP in the spinal cord following T-13 spinal cord hemisection in adult male Sprague-Dawley rats compared to that seen in sham controls. Spinal cords from both hemisected and sham control groups (N = 5, per time point) were examined on postoperative day (POD) 3, 5, 7, 14, and 108 following surgery. Sham operated rats displayed CGRP immunoreaction product in laminae I and II outer, Lissauer's tract, dorsal roots, and motor neurons of the ventral horn. In the hemisected group, densiometric data demonstrated an increased deposition of reaction product that was statistically significant, in laminae III and IV, both ipsilateral and contralateral to the lesion that extended at least two segments rostral and caudal to the hemisection site by POD 14, and remained significantly elevated as long as POD 108. Since upregulation alone of CGRP would occur in an acute temporal window (by 2 to 3 days following spinal injury), these results are interpreted to be invasion of laminae III and IV by sprouting of CGRP containing fine primary afferents. Intrathecal delivery of antibodies against purified 2.5S nerve growth factor for 14 days to the hemisected group resulted in CGRP density in laminae I through IV that was significantly less than that seen in untreated or vehicle treated hemisected groups and to sham controls. These data indicate changes in density and distribution of CGRP following spinal hemisection that can be manipulated by changes in endogenous levels of NGF. These observations suggest possible strategies for intervention in the development of various pain states in human SCI. PMID- 9344571 TI - Abnormalities in the development of the tectal projection from transplants of embryonic occipital cortex placed in the damaged occipital cortex of newborn rats. AB - We have examined the degree of precision in the topographic arrangement of the tectal projection developed by homotopic transplants of embryonic occipital cortex and tried to determine whether the development of the corticotectal projection is exclusively dependent on environmental cues or is also controlled by intrinsic factors. Transplants of embryonic (E16) occipital cortex were grafted into various areas of the occipital cortex (Oc1 or Oc2) of newborn rats and the organization of the tectal projection arising from the transplants was subsequently examined by injecting different neurotracers into the transplants. Our results indicate that in most cases the laminar and tangential distributions of the tectal projections from the transplants were abnormal. Indeed, whatever the location of the transplant in the host occipital cortex and whatever the placement of the injection into the transplant, a hybrid distribution of the tectal labeling was found, reminiscent of the pattern observed following tracer deposits in both Oc1 and Oc2 in intact animals. Since the grafts were composed of cells of both Oc1 and Oc2 embryonic origin, it is likely that the hybrid pattern of efferents reflects the heterogeneity of the embryonic origin of the cells composing the graft. These findings provide evidence that the development of the topographic distribution of neocortical efferents is not only dependent on factors extrinsic to the cortex and further indicate that even within one single cortical region, the occipital cortex, different areas (Oc1 vs Oc2) are not totally interchangeable. These findings might have important implications in transplantation experiments aiming at the reconstruction of damaged neocortical circuitry where a precise "point-to-point" reconstruction of the circuitry is expected. PMID- 9344573 TI - Tirilazad mesylate improves survival of rat and human embryonic mesencephalic neurons in vitro. AB - The survival rate of embryonic dopamine (DA) neurons after transplantation to the striatum is only 5-20%. Therefore, mesencephalic tissue from several donors needs to be implanted in a parkinsonian patient to induce a therapeutic improvement. Lazaroids are a group of neuroprotective compounds which inhibit lipid peroxidation. Previously, two lazaroids (U-74389G and U-83836F) have been found to improve the survival of both cultured and grafted rat DA neurons. The only lazaroid approved for human use is tirilazad mesylate. The objective of the present study was to explore the effects of tirilazad mesylate on DA neuron survival in cultures of rat ventral mesencephalon and its capacity to promote the in vitro cell viability of embryonic rat and human mesencephalic tissue, treated and dissociated in the same way as in clinical trials. After 7 days in vitro, the number of tyrosine hydroxylase-immunopositive, presumed DA neurons was 140% higher in rat cultures treated with 0.3 microM tirilazad mesylate than that in control cultures. Rat and human cell suspensions supplemented with tirilazad mesylate maintained a high degree of viability for several hours longer than control suspensions. These results indicate that tirilazad mesylate promotes the survival of both rat and human embryonic mesencephalic neurons in vitro. Tirilazad mesylate can be administered clinically and may become a useful tool for increasing survival of grafted DA neurons in patients, thereby reducing the needed quantity of human donor tissue. PMID- 9344572 TI - Transplantation of fetal neocortex ameliorates sensorimotor and locomotor deficits following neonatal ischemic-hypoxic brain injury in rats. AB - Ischemic brain injury in neonates can result in the degeneration of cortical and subcortical areas of brain and is associated with neurologic deficits. One approach to restoring function in conditions of ischemic brain injury is the use of neural transplants to repair damaged connections. This approach has been shown to reestablish neural circuitry and to ameliorate associated motor deficits in models of neonatal sensorimotor cortex damage. In this study, we utilized the Rice et al. rodent model of neonatal ischemic-hypoxic (IH) brain injury to assess whether transplantation of fetal neocortical tissue can promote functional recovery in tests of sensorimotor and locomotor ability throughout development and as adults. We show that animals that received neocortical grafts 3 days following the IH injury performed significantly better as adults on two measures of motor ability, the Rota-Rod treadmill and apomorphine-induced rotations, than did control animals that received sham transplants after the IH injury. Transplants were identifiable in 72% of the animals 10-12 weeks after implantation. Histochemical studies revealed that while the transplanted tissue did not establish normal cortical cytoarchitecture, cells and fibers within the grafts stained for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH d), choline acetyl transferase (ChAT), cholecystokinin (CCK), and glial fibrillary acidic protein (GFAP). These results suggest that transplantation of fetal neocortical tissue following IH injury in the neonatal period is associated with amelioration of motor deficits and that the grafted tissue demonstrated a neurochemical phenotype that resembled normal neocortex. This approach warrants continued investigation in light of potential therapeutic uses. PMID- 9344574 TI - GABA receptor agonist promotes reformation of the striatonigral pathway by transplant derived from fetal striatal primordia in the lesioned striatum. AB - Striatal lesions are known to cause the anterograde transneuronal degeneration of the substantia nigra pars reticulata (SNr) neurons in consequence to loss of GABAergic inhibitory striatonigral efferents. The present study was undertaken to examine whether long-term intraventricular administration of the GABA agonist muscimol could promote reformation of the striatonigral pathway arising from transplants by rescuing host SNr neurons from transneuronal death in rats with striatal ischemic lesions. Compared to nongrafted rats with striatal lesions, (i) a prominent axonal projection from the transplants to the ipsilateral substantia nigra, (ii) a significant increase in number of survived neurons in the ipsilateral SNr, and (iii) a significant reduction in number of apomorphine induced turning behaviors were found in grafted animals with muscimol infusion, but not in those without muscimol administration. These findings suggest that preservation of the host target neurons for grafted cells may increase an efficacy of cerebral implants in establishment of the host-graft fiber connections, possibly, leading to functional restoration. PMID- 9344575 TI - Effect of stereotaxic intrastriatal cografts of autologous adrenal medulla and peripheral nerve in Parkinson's disease: two-year follow-up study. AB - Studies in nonhuman primates with experimental parkinsonism have shown that intrastriatal cografts of autologous adrenal medulla and peripheral nerve yield greater behavioral improvement and graft survival than do adrenal medulla grafts alone. To test these observations, five patients with advanced Parkinson's disease were selected to receive unilateral intrastriatal adrenal medulla intercostal nerve cografts. They were evaluated using the Core Assessment Program for Intracerebral Transplantation (CAPIT) protocol. Three of these patients also underwent quantitative motor testing for the measurement of upper limb bradykinesia (movement time; MT). Following right flank adrenalectomy, cografts consisting of small fragments of adrenal medullary tissue and minced intercostal nerve were stereotaxically implanted into three targets in the right striatum using computerized tomography guidance. Surgery was uneventful and postoperative magnetic resonance imaging revealed accurate placement of the grafts. No morbidity was encountered. Results of 24 months of clinical and quantitative motor assessments postoperatively are reported. Total UPDRS motor scores in the "off" state improved from a mean preoperative score of 39.5 to 32.1 at 3, 29.7 at 6, 27.6 at 9, 28.5 at 12, 31.4 at 18, and 26.5 at 24 months after surgery. Total timed motor test scores during the "off" state improved 17.9% at 6, 23.3% at 9, 18.2% at 12, 38.2% at 18, and 34.9% at 24 months postoperatively compared to baseline. Movement time showed statistically significant improvement (repeated measures ANOVA, P < 0.05) in the left arm (contralateral to surgery) in all three patients tested. These results indicate that stereotaxic intrastriatal implantation of autologous adrenal medulla-peripheral nerve cografts can be performed safely and clinical improvement from this procedure is sustained for a period of 24 months. The clinical improvement was paralleled by improvement in objective, quantitative motor testing. PMID- 9344576 TI - Effect of flupirtine on Bcl-2 and glutathione level in neuronal cells treated in vitro with the prion protein fragment (PrP106-126). AB - Flupirtine, trade name Katadolon, is a centrally acting nonopioid analgesic that has recently been found to display cytoprotective activity in vitro and in vivo on neurons induced to undergo apoptosis. This report shows that the PrP106-126 fragment of the prion protein, which is the likely etiological agent for a series of encephalopathies, is toxic to cortical neurons in vitro. Simultaneously, PrP106-126 influences the molecular GSH content and the bcl-2 expression in neurons. Significant toxicity (32% reduction in cell viability) was observed at a concentration of 50 microM of the peptide after 9 days of incubation, while at higher concentrations toxicity increased to 70%. Neurotoxicity was greatly reduced following coincubation with 1 to 3 microg/ml flupirtine. Concomitant with PrP106-126-mediated cytotoxicity, glutathione (GSH) content fell by > 70% with respect to untreated controls. This decrease in GSH level was strongly blocked by flupirtine under incubation conditions that reduce cell toxicity. In addition to normalizing GSH content, flupirtine induced the expression of the anti apoptotically acting proto-oncogene bcl-2. Based on these in vitro data and on the favorable pharmacokinetic profile of the drug, we strongly suggest that flupirtine may prove useful for treatment of patients with prion disease. PMID- 9344578 TI - Editorial PMID- 9344577 TI - Expression of floor plate in dispersed mesencephalic cultures: role in differentiation of tyrosine hydroxylase neurons. AB - The availability of sufficient numbers of dopaminergic neurons for transplantation has been an important issue. Recently, it has been shown that the ventral floor plate (FP4-positive) cells and the transcription factor HNF-3beta are important in the signals that terminate proliferation and produce differentiation of the dopaminergic phenotype. In this study, dispersed mesencephalon from embryonic rats at Day 11 postcoitus (E-11), 1 day prior to the birth of TH cells, were cultured for 48 h and 1 week to evaluate TH neuronal differentiation and/or proliferation in vitro. The number of TH cells increased 14x between 48 h and 1 week in culture. In dispersed E-14 cultures, the presence of FP4 and HNF-3beta markers was demonstrated using immunohistochemistry. The majority of FP4-positive cell clusters were associated with TH neurons, suggesting that floor plate cells may have participated in TH neuron differentiation in culture. Antisense oligonucleotide probe for HNF-3beta mRNA added daily to cultured E-14 cells blocked the HNF-3beta expression, but had no effect on the FP4 or TH expression. These studies suggest a potentially important role for floor plate cells in the differentiation of TH cells, and differentiation and/or proliferation of TH cells in dispersed cultures of E-11 is demonstrated. PMID- 9344579 TI - The price of independence. PMID- 9344580 TI - The effect of the cyclin-dependent kinase inhibitor olomoucine on cell cycle kinetics. AB - The effect of the cyclin-dependent (CDK) inhibitors olomoucine and roscovitine on cell kinetics was studied. To this end, nonsmall cell lung cancer (NSCLC) cell line MR65 and neuroblastoma cell line CHP-212 were pulse labeled with bromodeoxyuridine (BrdUrd) and chased in culture medium, to which various concentrations of olomoucine or roscovitine were added. A dose-dependent inhibition of the G1/S-phase and G2/ M-/G1 transitions was observed. Furthermore, S-phase progression was also inhibited in a dose-dependent manner. Similarly, roscovitine, another CDK inhibitor with a 10-fold higher efficiency for both CDK1 and CDK2 as compared to olomoucine, showed the same effects at a 10-fold lower concentration. At the highest tested doses both olomoucine (200 microM) and roscovitine (40 microM) induced a complete cell cycle block in both cell lines, paralleled by the appearance of apoptotic figures. In these cultures a decrease in CDK1 protein level was found as shown by Western blotting. Bivariate CDK1/DNA analysis confirmed these observations and showed that a subpopulation of cells with characteristics of apoptosis became CDK1 negative. The presented data suggest that cyclins and CDKs are involved at an important nodal point shared by pathways regulating cellular proliferation and apoptosis. PMID- 9344581 TI - Inhibition of retinoic acid receptor function in normal human mammary epithelial cells results in increased cellular proliferation and inhibits the formation of a polarized epithelium in vitro. AB - The expression of retinoic acid receptor-beta (RAR beta) mRNA is absent or down regulated in a majority of breast cancers, suggesting that loss of retinoic acid receptor function may be a critical event in breast cancer carcinogenesis. We developed an in vitro system to investigate whether the loss of retinoic acid receptor (RAR) function might affect the proliferation and structural differentiation of normal cultured human mammary epithelial cells (HMECs). Utilizing a truncated retinoic acid receptor (RAR)-alpha construct exhibiting dominant-negative activity against retinoic acid receptor isoforms alpha, beta, and gamma (DNRAR), we inhibited normal retinoic acid receptor function in HMECs. Suppression of RAR function in HMECs resulted in reduced growth inhibition mediated by all-trans-retinoic acid (ATRA). Moreover, the doubling time of HMECs expressing the DNRAR was significantly shortened, associated with a decrease in the percentage of cells in G1 and an increase in the percentage of cells in S phase relative to controls. In addition, HMECs expressing the DNRAR cultured in prepared extracellular matrix exhibited a loss of extracellular matrix-induced growth arrest and formation of a polarized ductal epthelium. Our results suggest that ATRA and RARs may play an important role in regulating the proliferation of HMECs and in promoting differentiation. PMID- 9344582 TI - Acetylcholine receptor formation in mouse-chick chimera. AB - This study investigated possible interactions between motoneurons and somitic derived muscle cells in the formation of neuromuscular synapses in the myotome. The peculiarities of the neuromuscular synaptic pattern in chick and mouse embryos provided a model for studying the achievement of synaptogenesis between chick motoneurons and mouse muscle cells. In chick embryo, initial AChR clustering occurs well before innervation of the myotome, whereas in mouse embryo nerve axons invade the myotome extensively before the appearance of AChR clusters. Our approach was to replace somites from a chick host embryo with those derived from mouse donor embryos. We show that muscle cells from mouse myotome can differentiate in the chick embryo environment and form neuromuscular contacts with chick motor axons. Host axons invaded in ovo differentiating mouse myotome at a time when they had not yet reached the host myotome. This particular ingrowth of motor nerves was attributable to the mouse transplant since use of a quail somite did not produce the same effect as the mouse somite, which suggests that developing mouse muscles specifically modify the time course of chick axogenesis. The synaptic areas formed between chick motor axons and mouse myotubes developed according to the mouse pattern. Both the timing of their appearance and their morphology correlated perfectly with events in mouse synaptogenesis. These results indicate the important role played by postsynaptic membrane in controlling the first steps of AChR formation. PMID- 9344583 TI - Autofluorescence of live purple bacteria in the near infrared. AB - We have developed a novel microscope with which to study the fluorescence of cells in the near-infrared region (lambda = 750-2500 nm). For one of its first applications we report on the autofluorescence of live purple bacteria, Rhodospirillum rubrum, and suggest that the autofluorescent component is bacteriochlorophyll. The rapid fading of the autofluorescence of fixed bacteria and of purified bacteriochlorophyll suggests that the live bacteria are able to regenerate their pigment with a time constant of approximately 20 s. PMID- 9344584 TI - Involvement of p53 and WAF1/CIP1 in gamma-irradiation-induced apoptosis of retinoblastoma cells. AB - Retinoblastoma is an intraocular malignancy of childhood, which is very radiosensitive. gamma-irradiation, however, induces side effects, and the precise mechanisms of tumor cell death after the treatment remain unknown. In this study, we demonstrated that gamma-irradiation induced apoptosis (programmed cell death) in human retinoblastoma cell lines Y79 and WERI-Rb-1. The expression levels of p53, tumor suppressor gene product, and its-associated protein, WAF1/CIP1, were both up-regulated, and function of p53 was remarkably activated after the treatment. Moreover, apoptosis was induced in the absence of gamma-irradiation by overexpression of the WAF1/CIP1 gene in both retinoblastoma cells. These results indicate that the transfer of the WAF1/CIP1 gene may have potential for the treatment of human retinoblastomas instead of gamma-irradiation. PMID- 9344585 TI - Reduced epidermal expression of a PG-M/versican-like proteoglycan in embryos of the white mutant axolotl. AB - Axolotl embryos have previously been used to study neural crest cell migration. In embryos of the normal wild type, neural crest cells migrate subepidermally to form pigment cells. In the trunk of the white mutant embryo, these cells are unable to migrate, possibly due to an inherited delay in the maturation of the local extracellular matrix. The present investigation reveals a reduced incorporation of [35S]sulfate into PG-M/versican-like proteoglycans synthesized in epidermal explants from the dorsal trunk of white mutant embryos during stages pertinent to migration. This is the major form of proteoglycans in the subepidermal matrix, where they are assembled in large disulfide-stabilized supramolecular complexes. The reduction in [35S]sulfate incorporation is not due to qualitative differences between wild-type and white mutant proteoglycans but is paralleled by a reduced expression of mRNA for the core protein of the PG M/versican-like proteoglycan. We conclude that a reduced amount of these proteoglycans is produced by the white mutant embryo during the period critical for migration. PMID- 9344586 TI - Effects of intermittent or continuous gravitational stresses on cell-matrix adhesion: quantitative analysis of focal contacts in osteoblastic ROS 17/2.8 cells. AB - The relationship between cell morphology and cell metabolism and the role of mechanical load in bone remodeling is well known. Mechanical stimulation induces changes in the shape of osteoblasts, probably mediated by reorganization of focal contacts. We studied the influence of gravity (Gz) variations occurring during parabolic flight on osteoblast focal adhesion of ROS 17/2.8 osteosarcoma cells subjected to 15 or 30 parabolic flights. Significant flight-induced shape changes consisted of decreased cell area associated with focal contact plaque reorganization. Identical durations of continuous mechanical stress induced by centrifugation (2 Gz) or clinorotation (Gz randomization) had no major effect on cell focal adhesion. ROS 17/2.8 G2/M synchronization by treatment with nocodazole inhibited the flight-induced decrease in adhesion parameters. We concluded that ROS 17/2.8 cells are sensitive to Gz switches and that their adaptation is at least dependent on microtubule function. PMID- 9344587 TI - Alpha 6 beta 4 and alpha 6 beta 1 integrins associate with ErbB-2 in human carcinoma cell lines. AB - Growth factors modulate integrin-mediated cell adhesion and motility, and their receptors are thought to share proteins that mediate intracellular signaling with integrin receptors. The crosstalk between these receptors is thought to play a relevant role in transformation and tumor progression. To highlight possible interactions between growth factors and cell adhesion receptors we investigated whether integrins associate with tyrosine kinase receptors in tumor cells. By affinity chromatography and Western blot analyses of purified immune complexes, we studied the association of laminin receptors (alpha 6 beta 1 and alpha 6 beta 4) with ErbB-2 tyrosine kinase in human carcinoma cell lines. We demonstrated that the alpha 6 beta 4 and alpha 6 beta 1 integrins coprecipitated with ErbB-2 in lysates from carcinoma or NIH3T3 cells overexpressing ErbB-2. Integrin mediated activation of ErbB-2 receptors suggested that this association is functionally meaningful. Indeed, carcinoma cells treated with a monoclonal antibody to the alpha 6 integrin subunit showed a ligand-dependent increase of ErbB-2-phosphorylated molecules coprecipitated with integrins and an increased DNA synthesis. The interaction between growth factor receptors and integrins was also studied in NIH3T3 cells overexpressing alpha 6 beta 4 receptors and ErbB-2 protein. We report that cells overexpressing both receptors, but not those overexpressing a crippled ErbB-2, showed enhanced proliferation rates and invasiveness, further suggesting that alpha 6 beta 4 integrin and ErbB-2 receptor interaction might contribute to generate a more malignant phenotype in carcinoma cells. PMID- 9344588 TI - Microvilli elongate in response to hydrogen peroxide and to perturbations of intracellular calcium. AB - Using scanning electron microscopy and fluorescence microscopy, we have found that apical microvilli of diverse cell types, including nonepithelial cells, elongate in culture in response to the oxidative stress of hydrogen peroxide. The microvilli induced in culture on retinal pigment epithelial cells display a 30-nm axial periodicity similar to that described for stable microvilli of intestinal brush border. Microvilli can also be induced to elongate by chelating intracellular Ca2+ and by the Ca(2+)-uptake inhibitor thapsigargin. Thus a response of microvillar protrusion occurs widely and may be related to depletion of intracellular calcium stores. PMID- 9344589 TI - Stable transfection of U937 cells with sense or antisense RXR-alpha cDNA suggests a role for RXR-alpha in the control of monoblastic differentiation induced by retinoic acid and vitamin D. AB - Although retinoic acid (RA) has been known for many years to be a modulating agent that plays a role in generating both granulocytes and monocytes, the molecular mechanism underlying this role has not been defined in the monoblast lineage. In particular, the part played by the retinoid X receptors (RXRs), which are members of the steroid/thyroid hormone nuclear receptor family, has not been explored. In this study, therefore, the human monoblastic leukemia cell line U937 has been used as a model system to investigate the role of one of the RXRs, RXR alpha, in monoblast differentiation. RXR-alpha mRNA was present in untreated U937 cells, and levels increased after induction of differentiation with phorbol ester. The same was found for RXR-beta mRNA. Using plasmids containing sense or antisense RXR-alpha sequences under the control of an inducible promoter, we generated stably transfected cell lines which expressed either increased or decreased levels of RXR-alpha, respectively. The sense cell lines (U alpha S and its clonal derivative alpha G2S) showed increased sensitivity to RA, while the antisense cell lines (U alpha A and its clonal derivative alpha B5A) showed decreased sensitivity to RA, as demonstrated by growth inhibition and by regulation of an RA-responsive reporter gene. Both U alpha A and alpha B5A also failed to respond to another modulating agent, 1 alpha,25 dihydroxycholecalciferol (DHCC), but only U alpha S and not alpha G2S showed an enhanced response to DHCC. The combination of RA and DHCC together inhibited growth of both sense and antisense cell lines. In addition, alpha G2S exhibited increased expression of CD11b and CD54, while alpha B5A cells showed increased expression of CD102, suggesting that RXR-alpha has a role in regulating expression of cell adhesion molecules in U937 cells. These results demonstrate that RXR-alpha has a role in mediating growth inhibition and cell adhesion during myelomonocytic differentiation, and suggest that different species of heterodimers involving RXR-alpha may control the acquisition of different features of mature monocyte/macrophage function. PMID- 9344590 TI - The motility-associated proteins GAP-43, MARCKS, and CAP-23 share unique targeting and surface activity-inducing properties. AB - Local regulation of the cortical cytoskeleton controls cell surface dynamics. GAP 43 and MARCKS are two abundant cytosolic protein kinase C substrates that are anchored to the cell membrane via acyl groups and interact with the cortical cytoskeleton. Each of them has been implicated in several forms of motility involving the cell surface. Although their primary sequences do not reveal significant homologies, GAP-43, MARCKS, and the cortical cytoskeleton-associated protein CAP-23 (in the following, the three proteins will be abbreviated as GMC) share a number of characteristic biochemical and biophysical properties and an unusual amino acid composition. In this study we determined whether GMC may be related functionally. In double-labeling immunocytochemistry experiments GMC accumulated at unique surface-associated structures, where they codistributed. In transfected cells GMC induced the same range of characteristic changes in cell morphology and cell surface activities, including prominent blebs and filopodia. These activities correlated with local accumulation of transgene and had characteristic features of locally elevated actin dynamics, including loss of stress fiber structures, accumulation of beta-(cytosolic) actin at cell surface protrusions, and dynamic blebbing activity. Analysis of appropriate deletion and fusion constructs revealed that the surface accumulation pattern and cell surface activities were correlated and that minimal structural requirements included acylation-mediated targeting to the cell membrane and the presence of a predominantly GMC-type sequence composition. Based on these experiments and on the results of previous studies on GAP-43, MARCKS, and CAP-23, we propose that GMC may define a class of functionally related proteins whose local accumulation promotes actin dynamics and the formation of dynamic structures at the cell periphery. Superimposed on these general properties, differences in the regulation of membrane association and binding properties of effector domains would confer individual properties to each of these proteins. PMID- 9344591 TI - Effects of protein kinase C alpha overexpression on A7r5 smooth muscle cell proliferation and differentiation. AB - Smooth muscle cell differentiation and proliferation are increasingly seen to be intimately tied to the etiology of atherosclerosis and hypertension. To determine the role of PKC alpha in the regulation of smooth muscle cell differentiation and proliferation, the rat embryonic smooth muscle cell line A7r5 was transfected with an expression vector containing the full-length PKC alpha cDNA. Neomycin resistant clones which exhibited increased PKC alpha levels compared to wild-type cells were selected. The A7r5 cells overexpressing PKC alpha had altered morphology and decreased growth rates compared to wild-type cells and cells transfected only with the neomycin resistance gene. Electrophoretic mobility shift assays showed that nuclear extracts from overexpressing clones gave a different pattern of protein-DNA binding to an AP-1 consensus oligonucleotide compared to wild-type cells. In contrast to the growth characteristics of these clones, their levels of cell differentiation marker proteins such as vinculin and desmin were not affected by PKC alpha overexpression. Moreover, the smooth muscle specific differentiation marker alpha-actin was markedly reduced, while beta actin levels were found to remain unchanged. Northern blot analysis confirmed that alpha-actin downregulation occurred at the RNA level. Western blot analysis revealed that A7r5 cells have five different PKC isoforms and that these isoform protein levels were not changed by PKC alpha overexpression. These findings suggest that PKC alpha regulates growth and differentiation of A7r5 smooth muscle cells and that these changes might result from altered expression/function of AP 1 transcription factors. PMID- 9344592 TI - Selective monoclonal antibody recognition and cellular localization of an unphosphorylated form of connexin43. AB - A sequence-specific monoclonal antibody directed against the gap junction protein connexin43 (Cx43) is shown here to be specific for the unphosphorylated form of this protein. In tissues and cultured cells containing different phosphorylated and unphosphorylated forms of Cx43, the antibody detected only the latter as shown by Western blotting of native and alkaline phosphatase-treated samples. Immunohistochemically, this monoclonal antibody did not recognize gap junctions in the vast majority of cultured cardiac myocytes, where nearly all detectable Cx43 is phosphorylated. In contrast, it was able to detect some intracellular Cx43 in tracheal smooth muscle cells and an epithelial cell line (Cl-9 cells), producing patterns of labeling consistent with those seen using a polyclonal antibody that recognizes both phosphorylated and unphosphorylated forms of Cx43. Immunostaining of gap junctions in the cultured cells indicates that both phosphorylated and unphosphorylated Cx43 are present in some assembled gap junctions, suggesting that assembled junctions do not contain exclusively the phosphorylated form of the protein. Annular gap junctions, believed to form as part of the pathway for internalization and degradation of gap junctions, were only occasionally and sparsely labeled by the monoclonal antibody, indicating that complete protein dephosphorylation is not required for uptake and degradation of gap junctions. Furthermore, the ability of this antibody to recognize only unphosphorylated Cx43, and not any of the phosphorylated forms present in the tissues and cell types examined, suggests that a unique phosphorylation site, perhaps present in the epitope recognized by this antibody, must be phosphorylated prior to phosphorylation of Cx43 at other sites. PMID- 9344593 TI - Overexpression of antioxidant enzymes in transgenic mice decreases cellular ploidy during liver regeneration. AB - Reactive oxygen species (ROS) and antioxidant enzymes have been implicated in control mechanisms of cellular growth and proliferation. We investigated the influence of levels of endogenous antioxidant enzymes on liver regeneration in transgenic mice overexpressing human Cu,Zn-superoxide dismutase (SOD) and intracellular glutathione peroxidase (GP1) as a model system. After a two-thirds partial hepatectomy (PH), no significant difference was observed in rate of liver mass restoration among nontransgenic, SOD, and GP1 mice. In contrast, the level of polyploidization was significantly reduced in transgenic animals after PH, with a concomitant increase in 2N nuclei. The portion of 8N nuclei after 72 h reached 33.1, 15.8, and 22.1%, whereas the portion of 2N nuclei reached 7.5, 13.8, and 12.3% in nontransgenic, SOD, and GP1 mice, respectively. A similar effect was observed in another model of liver proliferation, during normal development around weaning time. Measurements of ROS production during PH indicate that overexpression of SOD leads to the decreased production of O2- and elevation of H2O2. Unexpectably, overexpression of GP in transgenic mice also results in increased production of H2O2 in hepatocytes. Finally, our data demonstrate that levels of endogenous antioxidant enzymes might influence the rate of hepatocyte polyploidization during liver proliferation. PMID- 9344595 TI - Reconstruction of peritoneal-like structure in three-dimensional collagen gel matrix culture. AB - The peritoneum is a serous membrane consisting of different kinds of cells and extracellular matrix components (ECM). The aim of the present study was to develop a three-dimensional (3D) in vitro culture system for possible investigation of pathological conditions of the peritoneum. Human omental mesothelial cells (MC) and endothelial cells from the umbilical vein (EC) were cultivated either on (MC) or in (EC) a preformed type I collagen matrix. In 3D culture mesothelial cells showed their phenotypical in vivo characteristics and the synthesis of a new basal membrane (BM). Endothelial cells developed vessel like structures, produce a BM and express E-selectin after TNF-alpha stimulation. This 3D culture system presents extended possibilities for analyzing mesothelial and endothelial cell behavior as well as the cell-cell and cell-matrix interactions involved in several pathological processes in the peritoneum. PMID- 9344594 TI - Localization of procollagen I in the lysosome/endosome system of human fibroblasts. AB - A significant amount of newly synthesized collagen is degraded intracellularly rather than secreted, but there is controversy about whether this process occurs in the lysosomes. We addressed this problem using confocal microscopy and immunofluorescence imaging to study the distribution of procollagen I in the Golgi and the lysosome/endosome system of cultured human fibroblasts. Cells were incubated under basal conditions and then permeabilized and exposed to fluorescently tagged probes for procollagen, Golgi markers (Helix pomatia binding protein or beta-coatamer protein), and lysosome/endosome markers (cathepsin B or LAMP-2). Strong signals for procollagen codistributed with the Golgi and lysosome/endosome markers. Of note, many structures were positive for procollagen and lysosome/endosome markers but not for Golgi markers. When cells were incubated with the proline analog cis-hydroxyproline, which inhibits correct triple helix formation and increases intracellular degradation, the amount of procollagen codistributing with the lysosome/endosome markers increased greatly. Similar results were obtained in I-cells, which do not have functioning lysosomal hydrolases. These findings strongly indicate that the lysosome/endosome system participates in the intracellular degradation of newly synthesized procollagen and that trafficking of procollagen to the lysosome/endosome system does not depend on the cells having active lysosomal hydrolases. We present a model that integrates our findings with other work and resolves inconsistencies in the literature. This model postulates the existence of three separate degradation paths for newly synthesized procollagen. In addition to the endosome/lysosome system, degradation also takes place in the proximal region of the secretory pathway such as the endoplasmic reticulum, cis-Golgi network, or cis-Golgi and in a distal region of the secretory pathway such as the trans-Golgi or trans-Golgi network. PMID- 9344596 TI - Neurofibromin colocalizes with mitochondria in cultured cells. AB - Mutations in neurofibromatosis type 1 target the gene coding for neurofibromin. While neurofibromin is able to accelerate the rate of GTP hydrolysis by cellular Ras proteins, its biological function is not well understood. To gain information regarding its function, the intracellular localization of neurofibromin was analyzed in cultured cell lines using polyclonal antisera raised against four neurofibromin-specific peptides, three from the carboxyl terminus and one from the amino terminus. In methanol-fixed cells distinct rod-like structures distributed throughout the cytoplasm were recognized by the antisera. Similar structures were seen with each antiserum, including affinity-purified antibodies, and in each of the cultured cell lines tested. Similar structures were seen in paraformaldehyde-fixed cells. Double staining experiments showed that these structures colocalize with mitochondria, but not with actin, beta-tubulin, or endoplasmic reticulum. When actin or tubulin structures within the cell were disrupted by separate antimitotic drugs, these stained structures retained their shape. Neurofibromin association with mitochondria was confirmed biochemically when highly purified mitochondrial fractions from bovine heart tissue were shown in Western analysis to contain neurofibromin. This association might be helpful in predicting identification of some of the cellular proteins with which neurofibromin interacts. PMID- 9344597 TI - Effects of cyclin D1 overexpression on G1 progression-related events. AB - In previous studies (W. Jiang, S. M. Kahn, P. Zhou, Y. (J. Zhang, A. M. Cacace, A. S. Infance, Y. Doi, R. M. Santella, and I. B. Weinstein 1993, Oncogene 8, 3447 3457) we reported that stable overexpression of cyclin D1 in R6 rat embryo fibroblasts shortens the G1 phase and impairs growth control. In the present study we examined the effects of cyclin D1 overexpression on other events involved in the G1 to S progression, utilizing the overexpressor cell line R6 ccnD1. We found that when compared to R6 control cells, serum-starved quiescent R6-ccnD1 cells had not only increased levels of the cyclin D1 protein but also increased levels of the cyclin E protein. The latter protein was complexed to phosphorylated cyclin-dependent kinase 2 (CDK2). However, in quiescent serum starved R6-ccnD1 cells this cyclin E-CKD2 complex lacked in vitro kinase activity due to the presence of a heat-stable inhibitory activity, apparently reflecting the inhibitory effects of the CDK inhibitors (CDKIs) p21WAF1 and p27KIP1. Serum stimulation of the quiescent R6-ccnD1 cells was associated with a loss of this inhibitory activity and a decrease in the levels of the latter two proteins, as the cells progressed through the G1 phase. On the other hand, serum stimulation of the control R6 cells was associated with both induction of cyclin E and increased levels of phosphorylated CDK2 proteins and decreased levels of p21WAF1 and p27KIP1, as the cells progressed through the G1 phase. Thus, even though overexpression of cyclin D1 can induce the expression of cyclin E and phosphorylated CDK2, premature activation of cyclin E-CDK2 kinase activity in quiescent cells or during progression through G1 appears to be blocked by CDKIs. Nevertheless, the R6ccnD1 cells have a shorter G1 phase than the control cells presumably due to the high levels of both cyclin D1 and cyclin E. Taken together, these results indicate that overexpression of cyclin D, which is frequently seen in human tumors, can have complex effects on the expression of other genes that control cell cycle progression. PMID- 9344598 TI - Thioredoxin expression and localization in human cell lines: detection of full length and truncated species. AB - Thioredoxin (Trx) is an intracellular multifunctional 12-kDa protein with a reduction/oxidation (redox) active disulfide constitutively expressed by most cells of the human body. Trx can also be released by cells such as lymphocytes upon activation or oxidative stress exposure and exert a cocytokine and cytoprotective activity. In addition, a truncated 10-kDa form of Trx has been reported. In order to better understand the function of full-length and truncated Trx, we have produced, for the first time, specific monoclonal antibodies, which can discriminate between the two forms. Using these novel antibodies, designated alpha Trx1 to alpha Trx4, a panel of cell lines derived from human B and T lymphocytes, monocytes, granulocytes, and melanomas was analyzed by immunochemical techniques. The cellular distribution differed between the two forms. All lines contained full-length Trx, also located to a minor extent on the cell surface. One exception was the melanoma cell line FM28.4, which did not show any Trx expression. Truncated Trx was present in most cells in minimal amounts only, whereas the monocytic cell lines THP-1 and U-937 expressed high amounts on the cell surface, as shown by flow cytometric analysis of living cells and confocal laser-scanning microscopy. The biological importance and function of the short versus long forms of Trx as detected by the antibodies are discussed. PMID- 9344599 TI - Acquisition of meiotic competence is related to the functionality of the phosphoinositide/calcium signaling pathway in the mouse oocyte. AB - The meiosis resumption process has been related to spontaneous cytoplasmic InsP3 dependent calcium oscillations in fully grown mouse oocytes. Our purpose was to determine whether the acquisition of meiotic competence during the growth phase of oogenesis was associated with that of Ca2+ oscillations and whether these oscillations were dependent on the phosphoinositide cycle. We used confocal laser scanning microscopy to image free calcium ions in fluo-3/AM-loaded oocytes recovered from 12- to 26-day-old mice for 15 min following follicular release. As expected, oocytes isolated from 12-day-old mice were totally incompetent to undergo GVB in vitro, whereas the GVB rate increased progressively with mouse age and oocyte diameter. The percentage of oocytes exhibiting spontaneous calcium oscillations and that of oocytes resuming meiosis were similarly correlated with the female age, with incompetent oocytes failing to exhibit spontaneous Ca2+ oscillations. It is noteworthy that regardless of the stage of growth, thapsigargin induced an ooplasmic calcium release from the InsP3-sensitive stores when it was added to the culture medium. However, intracytoplasmic microinjection of InsP3 induced a shorter sequence of Ca2+ oscillations in 12-day-old mouse oocytes than in 15-day-old mouse oocytes and, whereas InsP3 increased the GVB rate at 15 days, it was unable to induce GVB at 12 days. These data lead us to conclude that the acquisition of meiotic competence is related to the functionality of the InsP3 pathway and, correspondingly, to the oocyte's ability to generate spontaneous cytoplasmic InsP3-dependent calcium oscillations. PMID- 9344600 TI - Dynamic organization of DNA replication in one-cell mouse embryos: relationship to transcriptional activation. AB - We have analyzed the spatial and temporal relationship between transcription and replication sites during the first cell cycle in mouse embryos. Embryos were microinjected with both 5-bromouridine-5'-triphosphate and digoxygenin-11 deoxyuridine-5'-triphosphate to visualize transcription and replication sites respectively. We detected six different phases of replication during S phase and dated the onset of zygotic transcription at the end of the S phase. Using confocal microscopy, we showed that there is essentially no colocalization of replication and transcription sites at this stage of development. Moreover, studies on aphidicolin-treated embryos demonstrated that inhibition of DNA replication does not hinder transcriptional activation at the 1-cell stage. PMID- 9344601 TI - Laminin chain expression by chick chondrocytes and mouse cartilaginous tissues in vivo and in vitro. AB - We have observed that laminins are expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and adult mouse knee joint by the methods of in situ hybridization, immunohistochemistry, Western blotting, and immunoprecipitation. From in situ hybridization using similar sized RNA probes for different mouse laminin chains, mRNAs for the alpha 1, alpha 2, beta 1, beta 2, and gamma 1 chains were expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and the articular cartilage cap and epiphyseal growth plate of adult mouse knee joint. Through the use of chain-specific antibodies, staining for laminins was observed in the cytoplasm of chondrocytes from chick embryo sternum, mouse limb bud, and adult mouse knee joint. Western blot analysis confirmed the presence of laminin chains in the cells and sternal tissues. Cultured chick embryonic sternal chondrocytes expressed laminin mRNAs in the proliferating stage (2-3 days of culture) but the level increased in the aggregated cells during the maturation stage (5-7 days of culture). Comparable data were also obtained after immunostaining the cells. Thus, laminins are expressed in significant amounts by chondrocytes and may have an important role in cartilage development. PMID- 9344602 TI - Influence of cycloheximide-mediated downregulation of glucose transport on TNF alpha-induced apoptosis. AB - Enhancement of cellular sensitivity to TNF alpha-induced apoptosis by cycloheximide (CX) has been attributed to its quality as an inhibitor of protein synthesis, presumably by prevention of the synthesis of short-lived death antagonists. CX is also known to interfere with glucose transport, which in turn influences cell death. Hexose uptake, expression of glucose transporter (Glut) mRNAs and proteins, and other related factors were therefore examined upon induction of apoptosis with TNF alpha and CX in breast cancer cell lines. In the early phase of apoptosis, a dramatic decrease in glucose transport was observed, preceded by stimulation of Glut 1 and 3 mRNAs. Transport downregulation was also detectable upon incubation with CX alone, albeit to a lesser extent. With the doses used, TNF alpha had no such effect. Protein synthesis was inhibited to the same degree in TNF alpha/CX-treated apoptotic cells compared to viable CX-treated cells. Diminished hexose uptake was associated with decreased Vmax, while Glut affinity remained unaffected. As there was no evidence for changes in total cellular Glut content or for Glut translocation from the plasma membrane, a diminished intrinsic activity of Gluts must be postulated. In conclusion, CX is proposed to contribute to TNF alpha-induced apoptosis predominantly by interference with glucose transport; the exact nature of this effect remains to be elucidated. PMID- 9344603 TI - Susceptibility of human sperm to in situ DNA denaturation is strongly correlated with DNA strand breaks identified by single-cell electrophoresis. AB - Susceptibility of mammalian sperm DNA to low pH- or heat-induced denaturation in situ has shown very strong dose-response relationships with animal and human exposure to chemical and physical toxicants and also fertility potential. In this study, 23 human semen samples representing a wide range in percentage (7-86%) of sperm exhibiting abnormally high susceptibility of DNA in situ to denaturation were studied for the integrity of their DNA using alkaline comet assay (single cell microgel electrophoresis, pH 10.0). The percentage of comets observed for these samples ranged from 5 to 95%; these data correlated strongly with the percentage of sperm with increased DNA denaturability (r = 0.973; P < 0.001). Labeling of 3' ends of nicked DNA sites with 5-bromo-2'-deoxyuridine 5' triphosphate (BrdUTP) followed by tagging with FITC-BrdUTP monoclonal antibody and flow cytometry also indicated significantly strong correlations of BrdUTP incorporation with both abnormal susceptibility of DNA to denaturation (r = 0.859, P < 0.001) and comet assay (r = 0.812, P < 0.001). The relationship among susceptibility of sperm chromatin to acid denaturation in situ, BrdUTP incorporation, and formation of comets suggests that DNA fragmentation monitored by these assays may have important physiological relevance in terms of sperm quality and fertility potential. PMID- 9344604 TI - PKC regulation of microfilament network organization in keratinocytes defined by a pharmacological study with PKC activators and inhibitors. AB - The modulation by PKC activators and inhibitors of adhesion, spreading, migration, actin cytoskeleton organization, and focal complex formation in keratinocytes attaching to type I collagen was studied. Two actin microfilament networks, stress fibers and cortical actin, could be distinguished on the basis of cellular distribution and opposite regulation by growth factors, tyrosine kinase inhibitors, and PKC activators. Stress fiber formation was stimulated by growth factors and by PMA (100 ng/ml) and these stimulations were blocked by tyrosine kinase inhibitors (0.3 mM genistein and 1 microM herbimycin A). By contrast, the cortical network occurred in quiescent cells, was unaffected by tyrosine kinase inhibitors, and was broken down after PKC activation by PMA. Spreading, migration, and actin polymerization were completely blocked while adhesion efficacy was significantly decreased by three specific PKC inhibitors. Half-inhibition of migration was obtained with 0.025, 1, and 3 microM concentrations of calphostin C, chelerytrine chloride, and D-erythrosphingosine, respectively, which are concentrations close to those known to inhibit the PKC kinase function in vitro. Paxillin clustering, which was observed even in the presence of tyrosine kinase inhibitors, disappeared only when actin polymerization was completely impaired, i.e., in cells treated with PKC inhibitors or with both tyrosine kinase inhibitors and PMA, which indicated that focal complex formation was highly dependent on microfilament reorganization. The analysis of these data underscores a major regulation function of PKC in the molecular events involved in growth factor and adhesion-dependent regulation of microfilament dynamics. PMID- 9344605 TI - Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines. AB - Mature podocytes are among the most complex differentiated cells and possess a highly branched array of foot processes that are essential to glomerular filtration in the kidney. Such differentiated podocytes are unable to replicate and culturing of primary podocytes results in rapid growth arrest. Therefore, conditionally immortalized mouse podocyte clones (MPC) were established, which are highly proliferative when cultured under permissive conditions. Nonpermissive conditions render the majority of MPC cells growth arrested within 6 days and induce many characteristics of differentiated podocytes. Both proliferating and differentiating MPC cells express the WT-1 protein and an ordered array of actin fibers and microtubules extends into the forming cellular processes during differentiation, reminiscent of podocyte processes in vivo. These cytoskeletal rearrangements and process formation are accompanied by the onset of synaptopodin synthesis, an actin-associated protein marking specifically differentiated podocytes. In addition, focal contacts are rearranged into an ordered pattern in differentiating MPC cells. Most importantly, electrophysiological studies demonstrate that differentiated MPC cells respond to the vasoactive peptide bradykinin by changes in intracellular calcium concentration. These results suggest a regulatory role of podocytes in glomerular filtration. Taken together, these studies establish that conditionally immortalized MPC cells retain a differentiation potential similar to podocytes in vivo. Therefore, the determinative steps of podocyte differentiation and process formation are studied for the first time using an inducible in vitro model. PMID- 9344606 TI - Monitoring uptake of ellipticine and its fluorescence lifetime in relation to the cell cycle phase by flow cytometry. AB - Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazole) is a green fluorescent plant alkaloid that inhibits DNA topoisomerase II activity and possesses pharmacologic activity toward both murine and human leukemias in vivo. In this flow cytometric study, the uptake of ellipticine was monitored as a function of cell volume and cell cycle phase in viable human promyelocytic (HL-60) cells costained with the DNA fluorochrome Hoechst 33342. Uptake of ellipticine was time and dose dependent; however, drug content was quantitatively similar in all phases of the cell cycle when normalized for DNA content or similar to cell size when correlated with cell volume. The fluorescence lifetime values of ellipticine in HL-60 cells, as analyzed by novel flow cytometric analysis, reached a plateau when the intra-cellular ellipticine intensity was still rising with increasing drug concentration. Since the free drug and the different subcellular ellipticine complexes, including DNA and RNA, had different lifetime values, the changes in the lifetime values appear to reflect differing proportions of unbound drug to that bound to different cellular constituents in the cells. Further development of phase-sensitive flow cytometry will provide for multiple lifetime determinations so that quantitation of drugs bound to the different cellular components can be performed along with the simultaneous determination of total drug uptake and cell cycle position. Such analyses should provide useful information for the design of drugs with greater affinity for cytotoxic targets. PMID- 9344608 TI - Ultrastructural nucleolar alterations induced by an ametantrone/polyr(A-U) complex. AB - In the present study we examined the ultrastructural modifications as well as the precise distribution of DNA and RNA in RT4 cell nucleoli following a 3-h exposure to nontoxic or toxic doses of ametantrone (AMT), poly(adenylate-uridylate) (polyr(A-U), or an AMT/polyr(A-U) combination. While distribution of nucleic acids within the various nucleolar components is not modified following all treatments, the nucleoli exhibit several ultrastructural changes: redistribution of the nucleolar components, decrease in the number of fibrillar centers, and increase in the size of the fibrillar centers. The relative frequencies of the test agents to induce the apparition of nucleoli of compact type are AMT/polyr(A U) > AMT approximately polyr(A-U) > sham-treated, while the abilities of the test agents to induce the nucleolar segregation are AMT/polyr(A-U) approximately AMT > polyr(A-U) > sham-treated cells. These ultrastructural changes are characteristics of drugs that intercalate into DNA and inhibit rDNA transcription as well as rRNA processing and release of nascent preribosomes from the nucleolus. PMID- 9344607 TI - Modulation of heat-shock protein 70 (HSP70) gene expression by sodium butyrate in U-937 promonocytic cells: relationships with differentiation and apoptosis. AB - The administration of sodium butyrate at 0.75 mM induced the functional differentiation of U-937 human promonocytic leukemia cells with negligible cell mortality. However, the drug rapidly caused cell death with characteristics of apoptosis when used at concentrations of 5 mM and above. In addition, butyrate stimulated the expression of the stress-responsive heat-shock protein 70 (HSP70) gene when applied at both differentiation-inducing and apoptosis-inducing concentrations. The induction of HSP70 by butyrate was inhibited by the simultaneous addition of cAMP-increasing agents (dibutyryl cAMP or the combination of forskolin plus theophylline). However, these agents did not prevent differentiation and only partially reduced apoptosis. Moreover, the DNA topoisomerase II inhibitor etoposide, which provoked U-937 cell differentiation and apoptosis with the same or greater efficiency than butyrate, failed to stimulate HSP70 expression. Finally, it was observed that cAMP-increasing agents also abrogated the induction of HSP70 and reduced the apoptosis caused by cadmium chloride, a typical inducer of the stress response. Taken together, these results indicate that HSP70 expression is not required for differentiation of promonocytic cells, as earlier proposed, and that butyrate probably triggers the stress response in these cells. PMID- 9344609 TI - Prolactin inhibits EGF-induced DNA synthesis in mammary epithelium via early signaling mechanisms: possible involvement of protein kinase C. AB - Prolactin and prolactin agonists inhibited EGF-induced DNA synthesis in mammary epithelium, whereas other pituitary hormones had no effect on EGF-induced DNA synthesis. The inhibitory effect of prolactin was seen for EGF and TGF-alpha, but not for IGF-I or cholera toxin. Autoradiography indicated that prolactin decreased the ability of EGF to induce cells to progress to S phase of the cell cycle, and time course studies indicated that the effects of prolactin were not due to an altered timing of DNA synthesis induction. Prolactin addition within 30 min of adding EGF was necessary to inhibit EGF-induced DNA synthesis. Conditioned media from prolactin-treated cells from which prolactin had been neutralized with the extracellular domain of the prolactin receptor had no effect on EGF-induced DNA synthesis, suggesting that the effect was due to prolactin, not an autocrine factor induced by prolactin. Prolactin induced a rapid association of protein kinase C with the membrane fraction of NMuMG cells, as well as increased threonine phosphorylation of the EGF receptor. Protein kinase C inhibitors eliminated most of the inhibitory effect of prolactin on EGF-induced DNA synthesis. The protein kinase C inhibitor Calphostin C restored high-affinity EGF binding in prolactin-treated cells and reversed the inhibitory effect of prolactin on EGF-induced EGF receptor tyrosine phosphorylation. PMID- 9344610 TI - TGF-alpha-induced breakdown of stress fibers and degradation of tropomyosin in NRK cells is blocked by a proteasome inhibitor. AB - Treatment of NRK cells with TGF-alpha in the presence of serum initiates disassembly of cytoskeletal stress fibers and suppresses the synthesis of tropomyosin isoforms (TMs) 1, 2, and 3 but not TMs 4 and 5 (Cooper et al., Cancer Res. 47, 4493-4500, 1987). In order to determine how the loss of tropomyosin is induced and what role it plays in cytoskeletal disruption, the turnover of tropomyosin was studied in the presence of the transforming growth factor and protease inhibitors. Cells were pulse-labeled with [35S]methionine and chased in the absence or the presence of the growth factor. It was found that TMs 1, 2, and 3 are degraded at about twice the rate of TMs 4 and 5 in control cells and that the rate of degradation of TMs 1-3 is accelerated by the growth factors. Degradation of TMs in control and growth factor-treated cells is blocked by a membrane-permeable inhibitor of cysteine proteases (LLnL) that acts upon calpains and proteasomes, and the cells maintain a flattened shape with a normal complement of stress fibers. Application of inhibitors that block calpains but not proteasomes does not block TM degradation. Treatments (suspension culture or cytochalasin B) that disrupt stress fibers without application of the growth factors also accelerate TM degradation, suggesting that acceleration of TM degradation is a consequence of its release from stress fibers during their breakdown. The normally more rapid turnover of the TM isoforms 1-3 that are lost in the phenotypically transformed cells could serve to facilitate the cytoskeletal reorganization that follows the activation of signal transduction pathways by the transforming growth factors observed in this study or during other rearrangements of the cytoskeleton such as occur during cell migration or mitosis. PMID- 9344611 TI - Glucocorticoids stimulate growth of human papillomavirus type 16 (HPV16) immortalized human keratinocytes and support HPV16-mediated immortalization without affecting the levels of HPV16 E6/E7 mRNA. AB - We investigated the effects of the glucocorticoids hydrocortisone and dexamethasone on human papillomavirus type 16 (HPV16)-mediated human cell carcinogenesis using normal human keratinocytes (HKc) and HKc immortalized by transfection with HPV16 DNA (HKc/HPV16). Normal HKc did not require glucocorticoids for proliferation. In contrast, growth of early passage HKc/HPV16 strictly required these hormones, although glucocorticoid dependence became less stringent during in vitro progression. Glucocorticoid dependence was acquired by HKc early after immortalization with HPV16 DNA, and glucocorticoids were required for efficient HKc immortalization. However, treatment of HKc/HPV16 with hydrocortisone or dexamethasone did not increase the steady-state levels of HPV16 E6/E7 mRNA or protein. Firefly luciferase activity expressed under the control of the HPV16 upstream regulatory region and P97 promoter increased by about fourfold following dexamethasone treatment of HeLa, but only twofold in HKc/HPV16, and less than twofold in SiHa. However, all of these cell lines expressed sufficient endogenous glucocorticoid receptors to allow for a dexamethasone response of the mouse mammary tumor virus promoter. These results indicate that mechanisms other than a direct influence by glucocorticoids on HPV16 early gene expression may contribute to the striking biological effects of these steroids on HPV16-mediated human cell carcinogenesis. PMID- 9344612 TI - A complete epithelial organization of Caco-2 cells induces I-FABP and potentializes apolipoprotein gene expression. AB - The culture of Caco-2 cells on plastic support impairs the expression of several genes involved in lipid metabolism. We describe culture conditions that permit the expression of the I-FABP gene and better expression of the apolipoprotein A I, C-III, and A-IV genes. Basal lamina deposited on filters as well as the nature of nutrients on the apical side differentially modulated mRNA expression of I FABP, APOBEC-1, and apolipoprotein genes. Growing cells on a filter led to functional polarization, illustrated by a secretion of apo B at the basal side, which induced the expression of the I-FABP, APOBEC-1, and apo A-IV genes and highly increased the expression of the apo C-III gene. Moreover, basal lamina deposited on the filter enhances the mRNA expression of apo A-I. Apo C-III and A IV mRNA levels were decreased when cells were grown on a filter covered with basal lamina in the presence of a medium deprived of protein and lipid on the apical side, whereas these conditions had no effect on I-FABP, apo A-I, and APOBEC-1 mRNA levels. The addition of lipid micelles on the apical side had various effects, according to the genes. Caco-2 cells cultured under the conditions described here closely resembled enterocytes and represent a useful tool for studying the regulation of genes involved in lipid metabolism. PMID- 9344613 TI - Prostaglandin H synthase expression is variable in human colorectal adenocarcinoma cell lines. AB - The expression of prostaglandin H synthases can be induced by many stimuli and is likely to be important in control of the cell cycle. The analysis of prostaglandin H synthase-1 and -2 expression in colon adenocarcinoma cell lines is a useful model system for studying the function of the prostaglandin H synthases, especially with regard to proliferation and adhesion. Prostaglandin H synthase-1 protein is not found in any of eight human colon adenocarcinoma cell lines. Expression of prostaglandin H synthase-2 is variable for the eight cell lines: three constitutively expressed active protein, four did not express this gene at all, and one had mRNA but no active protein. Thus, five colorectal adenocarcinoma cell lines exhibit "null" expression of prostaglandin synthase-2. The three cell lines with constitutive expression of prostaglandin H synthase-2 produce PGE2. Prostaglandin E2 production could be inhibited by aspirin and NS398 without inhibiting proliferation, while direct addition of prostaglandin E2 inhibits proliferation. Adhesion to collagen IV and fibronectin was stronger in those cell lines that expressed prostaglandin H synthase-2. The constitutive expression of prostaglandin H synthase-2 is associated with increased adhesion to extracellular matrix components and a potential inhibition of proliferation through the production of prostaglandin E2. The absence of PGH synthase-2 expression in some cell lines may result from the original tumor's need to inactivate these associated functions. Our evidence suggests that PGH synthase-2 is a possible candidate for a tumor suppressor gene at 1q23-qter. PMID- 9344614 TI - 1-(Carboxymethylthio)tetradecane attenuates PDGF- and TPA-induced c-fos mRNA expression and increases the formation of phosphatidylethanolamine with a shift from less to more polar molecular species in C3H/10T1/2 cells. AB - 1-(Carboxymethylthio)tetradecane caused C3H/10T1/2Cl8 and C3H/10T1/2Cl16 to incorporate 10 times more [32P]Pi into diacylphosphatidylethanolamine than control. This 3-thia fatty acid caused a shift in incorporation of 32P radioactivity into phosphatidylethanolamine species from species with long to species with short HPLC elution times. The increase in 32P-labeling was parallelled by a change in the apparent mass of phosphatidylethanolamine to a higher proportion of molecular species with short elution times than with long elution times. 1-(Carboxymethylthio)tetradecane caused loss of molecular species containing stearoyl groups. These results indicate that culturing the cells with 1-(carboxymethylthio)tetradecane causes looser packing and an increase in fluidity of the diacylphosphatidylethanolamine molecules in the membranes. 12-O tetradecanoyl phorbol-13-acetate (TPA), platelet-derived growth factor (PDGF)-BB, or PDGF-AA stimulation of 1-(carboxymethylthio)tetradecane-treated cells resulted in decreased maximal levels of c-fos mRNA expression, indicating attenuation of signal transduction. Compared to cells not treated, the levels of both PDGF-alpha and PDGF-beta receptors were lower while GTPase-activating protein and phospholipase C-gamma levels were not altered in C3H/10T1/2Cl8 and C3H/10T1/2Cl16 cells cultured in the presence of 1-(carboxymethylthio)tetradecane. Our data demonstrate that 1-(carboxymethylthio)tetradecane-mediated changes in phospholipid structure and composition may affect PDGF- and TPA-mediated c-fos gene regulation in fibroblasts. PMID- 9344615 TI - Cleavage of beta 4 integrin by matrilysin. AB - Overexpression of the matrix metalloproteinase matrilysin and the absence of beta 4 integrin are two features characteristic of human prostate carcinoma. In the following study we demonstrate that the beta 4 integrin, but not the alpha 6 or beta 1 integrin subunits, is cleaved by matrilysin in vitro. A specific fragment of 90 kDa is generated using matrilysin, which is not observed with other proteases. Two putative cleavage sites for matrilysin within the extracellular domain of the beta 4 integrin at residues 107 (isoleucine, prior to the ligand binding region) and 417 (leucine, prior to cysteine-rich region) are identified by sequence comparisons with known matrilysin substrates. The selective cleavage of the beta 4 integrin by matrilysin may partly explain the loss of beta 4 integrin expression in invasive prostate carcinoma. PMID- 9344616 TI - In vivo analysis of nuclear protein traffic in mammalian cells. AB - We have developed an experimental system to study nucleocytoplasmic traffic of proteins in living mammalian cells. Toward this goal, substrates were generated that contain several copies of Aequorea victoria green fluorescent protein (GFP). To follow facilitated transport across the nuclear envelope we created reporter proteins that carry different nuclear localization sequences (NLSs). The expression of reporter genes was controlled by an inducible promoter. Transiently transfected HeLa cells were employed to follow the sorting of fluorescent reporter proteins. When NLS-GFP fusions were located in HeLa cells, we found that direct fusion of the NLS derived from SV40 T-antigen to GFP prevented nuclear accumulation of the protein. However, insertion between NLS and GFP of different linkers encoding small amino acid residues produced reporter proteins that were competent for nuclear import. Taken together, we have generated unique tools for the characterization of nuclear protein import in dividing mammalian cells. PMID- 9344617 TI - G1-phase arrest is not a prerequisite for encystment in Physarum. AB - Amoebae of the Myxomycete Physarum polycephalum form resistant, walled cysts when the food bacteria in a culture have been consumed. No G1 phase has been detected in the vegetative amoebal cell cycle, most of which comprises the G2 phase. Mature cysts are also in G2, but it has been reported that a G1 phase of roughly 24 h, followed by an S phase, is obligatory prior to encystment. We used flow cytometry to determine the distribution of DNA contents in amoebal cultures at intervals during vegetative growth and encystment. In all cultures, the cells were predominantly in G2 phase, and the percentage of cells with G1 DNA content remained very low. We conclude that an extended G1 phase of 24 h did not occur in our cultures and cannot be a prerequisite for encystment. PMID- 9344618 TI - Different kinetics of senescence in human fibroblasts and peritoneal mesothelial cells. AB - Senescence has been reported for a wide variety of human cell types. In cultures of human fibroblasts the process is due to a percentage of the cells becoming senescent at each passage rather than all the cells entering senescence simultaneously at the end of the life span. By measuring the percentage of fibroblasts which are still cycling at each passage, a rate of decline in the growth fraction, which mirrors the rate of senescence, can be obtained. However, such an analysis has never been undertaken in multiple cell types using the same method to identify cycling cells. It is thus unknown if the rate of senescence is the same or different in cultures of different human cell types. To answer this question the rates of decline in the cycling fractions were simultaneously measured in two cultures of human cells (AGO7086A, peritoneal mesothelial cells; and 2DD, human dermal fibroblasts) which have practically identical in vitro life spans. 2DD fibroblasts showed a rate of decline of 0.89% cycling cells per population doubling when the data obtained were fitted to a simple linear equation. However, AGO7086A gave a decline of approximately 2.2% per population doubling. Thus mesothelial cells enter senescence significantly faster than fibroblasts (P < 0.001). This decline in the growth fraction was accompanied by an increasing fraction of mesothelial cells which retained detectable endogenous beta-galactosidase activity at pH 6. Such activity has previously been shown to be associated with senescent human fibroblasts. These findings suggest that the process of senescence has common features in different cell lineages but that the rate of the process can differ markedly between them. PMID- 9344619 TI - In Memoriam: HUGH DAVSON 1909-1996 AB - No abstract Copyright 1996 Academic Press Limited PMID- 9344620 TI - Risk characterization: A bridge to informed decision making. AB - Regulatory decisions should be made in the most expert and informed way since they are precipitated by real and perceived threats to human health, under the glare of public scrutiny. In 1994, the National Research Council (NRC) reported that the U.S. Environmental Protection Agency's (USEPA's) overall approach to assessing risks is fundamentally sound, but the Agency must more clearly establish the scientific and policy basis for risk estimates and better communicate the associated uncertainties. On March 21, 1995, USEPA issued a risk characterization policy and guidance. In this policy, an effective risk characterization must fully and clearly characterize risks and disclose the scientific analysis, uncertainties, assumptions, and science policy that underlie decisions throughout the risk assessment process. A number of regulatory reform bills which required risk characterization as part of all Federal risk assessments were introduced by the 104th Congress. The purpose of this workshop was to familiarize Society of Toxicology members with: (1) key elements to be considered in risk characterization and (2) new advances in risk characterization addressed by Federal and State agencies, industry, academia, NRC, and Presidential/Congressional Commission on Risk Assessment and Risk Management. Furthermore, the main objective was to engage the audience in discussing the proper role of science in risk assessment-risk management interface to make informed decisions in the face of scientific uncertainty. PMID- 9344621 TI - Toxicity of divinylbenzene-55 for B6C3F1 mice in a two-week inhalation study. AB - Divinylbenzene (DVB) is a crosslinking monomer used primarily for copolymerization with styrene to produce ion-exchange resins. The toxicity of inhaled DVB was investigated because of the potential for worker exposure and the structural similarity of DVB to styrene, a potential carcinogen. Male and female B6C3F1 mice were exposed to 0, 25, 50, or 75 ppm DVB for 6 hr/day, 5 days/week for up to 2 weeks. Six mice/sex/dose group were killed after 3, 5, and 10 exposures and six mice/sex in the 75 ppm group were killed 7 days after 10 exposures. The most severe effects occurred in the nasal cavity and liver, with less severe effects occurring in the kidneys. In the nasal cavity olfactory epithelium acute necrosis and inflammation were present at early time points followed by regeneration, architectural reorganization, and focal respiratory metaplasia by 7 days after the last exposure. Olfactory epithelial changes were concentration-dependent with extensive involvement at 75 ppm and peripheral sparing at 25 ppm. There was also necrosis and regeneration of olfactory associated Bowman's glands as well as the lateral nasal (Steno's) glands. Hepatocellular centrilobular (CL) necrosis was observed only in the 75 ppm dose group and was similar to that caused by styrene. A time-dependent progression was observed, characterized by CL degeneration after 1 exposure, necrosis after 3 and 5 exposures, and chronic inflammation with CL karyomegaly after 10 exposures and 7 days after the 10th exposure. Hepatic GSH levels were decreased in a dose dependent manner. In the kidneys, transient tubular damage was observed in some male mice exposed to 75 ppm, and appeared to be a response to DVB-induced tubular epithelial injury. PMID- 9344622 TI - Coincubation of rat renal proximal tubules with hepatic subcellular fractions potentiates the effects of para-aminophenol. AB - Treatment of rats with para-aminophenol (PAP) (300 mg/kg ip) produced decreases in renal nonprotein sulfhydryl (NPSH) content, oxygen consumption, and adenine nucleotide concentrations 2-4 hr following administration. In contrast, incubation of rat renal tubules with up to 1 mm PAP for 4 hr produced inconsistent changes in renal tubules. This discrepancy suggested that extrarenal metabolism of PAP may be involved in PAP bioactivation and nephrotoxicity. We designed the present studies to test the hypothesis that hepatic metabolism of PAP potentiates the effects of PAP on renal tubules. Incubation of renal tubules with 0.5 mm PAP and 10 mg protein from hepatic postmitochondrial supernatant (S9 fraction) in the absence of glutathione (GSH) for 4 hr did not alter renal oxygen consumption or adenine nucleotide metabolite concentrations. We observed no changes when we incubated tubules with 0.5 mm PAP and 1 mm GSH in the absence of hepatic S9 fraction. However, incubation of renal tubules with 0.5 mm PAP, 1 mm GSH, and 10 mg hepatic S9 protein for 4 hr significantly decreased renal oxygen consumption and adenosine triphosphate and total nucleotide concentrations. These data suggest that the effects of PAP in renal tubules may be potentiated by enzymatic metabolism of PAP, possibly involving oxidation and GSH conjugation. From experiments using hepatic microsomes or cytosol instead of S9 fraction, we found that changes were produced when we incubated tubules with PAP in the presence of hepatic microsomes, but not cytosol. These data suggest that hepatic microsomal metabolism of PAP may contribute to the production of changes in renal tubules in vitro. PAP-induced changes in renal proximal tubules were prevented when we included a beta-nicotinamide adenine dinucleotide phosphate (NADPH) generating system in the incubation medium. The protective effect of NADPH persisted when microsomes were inactivated by incubation with 1 aminobenzotriazole, a cytochrome P450 inhibitor. These data suggest that cytochrome P450-dependent oxidation is not involved in the production or prevention of PAP-induced changes in renal tubules. The enzyme(s) responsible for PAP bioactivation and the mechanism(s) by which NADPH protects renal tubules from PAP-induced decrements in oxygen consumption and adenine nucleotide concentrations are currently unclear. PMID- 9344623 TI - Use of sequentially administered stable lead isotopes to investigate changes in blood lead during pregnancy in a nonhuman primate (Macaca fascicularis). AB - The effects of pregnancy on the flux of lead from maternal bone were investigated in five females from a unique colony of cynomolgus monkeys (Macaca fascicularis) which had been dosed orally with lead (approximately 1100-1300 microg Pb/kg body wt) throughout their lives (about 14 years). Through the use of stable lead isotopes 204Pb, 206Pb, and 207Pb, it was possible to differentiate between the lead contributed to blood lead from the skeleton and the lead contributed from the current oral dose. Blood samples and bone biopsy samples taken before, during, and after pregnancy were analyzed for lead (total and stable isotope ratios) by thermal ionization mass spectrometry. Through the use of end-member unmixing equations, the contribution to blood of lead from maternal bone during pregnancy was estimated and compared to the contribution of lead from maternal bone before pregnancy. A 29 to 56% decrease in bone lead mobilization in the first trimester was followed by an increase in the second and third trimesters, up to 44% over baseline levels. In one monkey, the third-trimester increase did not reach baseline levels. In a single low-lead monkey, a similar decrease in the first trimester was followed by a 60% increase in the third trimester, indicating that a similar pattern of flux is seen over a wide range of lead concentrations. Analysis of maternal bone and fetal bone, brain, liver, and kidneys confirmed a substantial transplacental transfer of endogenous lead. Lead concentrations in fetal bone often exceeded maternal bone lead concentrations. From 7 to 39% of the lead in the fetal skeleton originated from the maternal skeleton. PMID- 9344624 TI - Physiologically based pharmacokinetics and the dermal absorption of 2 butoxyethanol vapor by humans. AB - It has generally been assumed that the skin contributes only minor amounts to the total uptake of solvent vapors, relative to the respiratory tract. Contrary to this assumption, the widely used glycol ether solvent, 2-butoxyethanol (BE), has been reported to be more effectively absorbed through the skin (75% of the total uptake) than through the lungs of humans (Johanson and Boman, 1991, Br. J. Ind. Med. 48, 788). The possibility that the finger prick blood sampling technique used in the Johanson and Boman study was confounded by locally high concentrations of BE at the site of absorption was suggested using a previously developed PBPK model (Corley et al., 1994, Toxicol. Appl. Pharmacol. 129, 61). The current study was conducted to verify the PBPK analysis and to determine whether or not the skin was the major site for absorption of BE vapor by exposing one arm from each of six human volunteers to 50 ppm 13C2-BE vapor for 2 hr. To evaluate the potential consequences of blood sampling techniques, samples were taken from both the unexposed arm (catheter; during and after exposure) and the exposed arm (finger prick; end of the exposure only) for analysis of both BE and its major metabolite, butoxyacetic acid (BAA). Butoxyacetic acid is responsible for the hemolysis observed in toxicity studies with laboratory animals. Humans, however, are significantly less sensitive to this effect. The concentration of BE in the finger prick blood samples averaged 1500 times higher than the corresponding concentration in venous blood sampled from a catheter installed in the unexposed arm at the end of the exposure. Blood BAA levels were generally within a factor of 4 of each other for the two techniques and, therefore, was considered a better indicator of systemic absorption. Urine was collected for 24 hr and analyzed for the following metabolites found in rat metabolism studies: free and conjugated BE, BAA, ethylene glycol (EG), and glycolic acid (GA), with only BAA detected in the human urine. More importantly, urinary BAA was found to be extensively conjugated ( approximately 67%) with glutamine, confirming recent reports. These results, coupled with PBPK modeling of worst-case exposure scenarios (no clothing, 100% of the body was exposed), demonstrated that no more than 15-27% (low-to-high relative temperatures and humidities), not 75%, of the total uptake of BE could be attributed to the skin of humans during simulated 8 hr exposures to the ACGIH TLV concentration of 25 ppm. Even less of the total uptake was attributed to the skin during simulations of exercise with whole-body exposures (5-9%) or by more realistic exposures of only the arms and head (1-8%). As a result, humans are unlikely to reach hemolytic concentrations of the metabolite BAA in blood following vapor exposures to BE. PMID- 9344625 TI - Dose response for the stimulation of cell division by caffeic acid in forestomach and kidney of the male F344 rat. AB - Caffeic acid (CA, 3,4-dihydroxycinnamic acid), at 2% in the diet, had been shown to be carcinogenic in forestomach and kidney of F344 rats and B6C3F1 mice. Based on its occurrence in coffee and numerous foods and using a linear interpolation for cancer incidence between dose 0 and 2%, the cancer risk in humans would be considerable. In both target organs, tumor formation was preceded by hyperplasia, which could represent the main mechanism of carcinogenic action. The dose response relationship for this effect was investigated in male F344 rats after 4 week feeding with CA at different dietary concentrations (0, 0.05, 0.14, 0.40, and 1.64%). Cells in S-phase of DNA replication were visualized by immunohistochemical analysis of incorporated 5-bromo-2'-deoxyuridine (BrdU), 2 hr after intraperitoneal injection. In the forestomach, both the total number of epithelial cells per millimeter section length and the unit length labeling index of BrdU-positive cells (ULLI) were increased, about 2.5-fold, at 0.40 and 1.64%. The lowest concentration (0.05%) had no effect. At 0.14%, both variables were decreased by about one-third. In the kidney, the labeling index in proximal tubular cells also indicated a J-shaped (or U-shaped) dose response with a 1. 8 fold increase at 1.64%. In the glandular stomach and in the liver, which are not target organs, no dose-related effect was seen. The data show a good correlation between the organ specificity for cancer induction and stimulation of cell division. With respect to the dose-response relationship and the corresponding extrapolation of the animal tumor data to a human cancer risk, a linear extrapolation appears not to be appropriate. PMID- 9344626 TI - Comparison of olestra absorption in guinea pigs with normal and compromised gastrointestinal tracts. AB - Female guinea pigs (12/group) were given a single dose of [14C]olestra by gavage after consuming either 3% poligeenan in tap water (Compromised group) or just tap water (Normal group) for 5 weeks. A Sentinel group (N = 2) was given 3% poligeenan for 5 weeks. Ten sentinel animals were killed 1 day before and 10 1 day after the other animals were dosed with [14C]olestra and their gastrointestinal tracts were examined by histology. The Compromised and Normal animals were endoscoped just before dosing with [14C]olestra. Urine and feces were collected continuously and CO2 was collected for 7 days after dosing. The samples were analyzed for 14C and urine was also analyzed for [14C]sucrose. Animals (3/group) were killed 1, 3, 7, and 21 days after dosing, and tissues were collected and assayed for 14C. Tissue lipids were extracted, fractionated by high pressure liquid chromatography, and analyzed for [14C]olestra by liquid scintillation. Animals fed poligeenan showed mucosal edema, congestion, ulceration, and fibrin deposition within the distal colon and rectum. Histology revealed inflammation, epithelial degeneration, and multifocal ulceration of the cecum, distal colon, and rectum. The gastrointestinal mucosae of nonpoligeenan fed animals were normal. No [14C]olestra was detected in liver lipids and no [14C]sucrose was found in the urine for any animal in the Normal or Compromised groups, indicating that intact olestra was not absorbed. The amount, distribution, and elimination of absorbed 14C did not differ between guinea pigs with normal and compromised gastrointestinal tracts. The poligeenan-treated animals displayed mucosal damage similar to that seen in human inflammatory bowel diseases; therefore, these results suggest that patients with inflammatory bowel conditions will not absorb olestra to any greater extent than normal healthy people. PMID- 9344627 TI - The duplication cycle in Aspergillus nidulans. AB - The duplication cycle encompasses the spectrum of events required for the growth and division of individual cells within a fungal hyphae. Recent advances in understanding the mechanisms which underlie nuclear division and cellular morphogenesis in the filamentous fungus Aspergillus nidulans have shown that in many respects, the duplication cycle differs significantly from the cell cycles of both budding and fission yeast. The purpose of this review is to summarize these advances and to highlight the fundamental differences between the duplication cycle and the yeast cell cycles. In addition, it is argued that the duplication cycle is controlled by cellular regulatory networks which integrate the processes of nuclear division, cellular morphogenesis, and cell growth with each other. Functional dissection of these networks should help to reveal features that are unique to the hyphal mode of growth. PMID- 9344628 TI - Complementation analysis of carotenogenic mutants of Mucor circinelloides. AB - Mutants of the filamentous fungus Mucor circinelloides altered in the synthesis of beta-carotene (genotype car) have been isolated by direct inspection of the color shown by the colonies derived from mutagenized spores. The mutants were analyzed for the carotenoid content in darkness and light and studied with respect to complementation in heterokaryons made by spheroplast fusion. The results revealed the existence of at least four complementation groups. The carB mutants are white and accumulate phytoene. The carR mutants are red and accumulate lycopene. The carP mutants are most likely disturbed in the synthesis of phytoene: two of them are white and do not accumulate carotenoids; another carP mutant is yellowish, probably because it is leaky. There are three yellowish mutants which belong to one or more complementation groups different from carB, carR, and carP. Two white mutants obtained in a single mutagenization step failed to complement with the carR and the carP mutants. The wild-type and the carB and carR mutants tested for M. circinelloides showed similar photoinduction. PMID- 9344629 TI - Null alleles of creA, the regulator of carbon catabolite repression in Aspergillus nidulans. AB - CreA is the major regulatory protein involved in carbon catabolite repression in Aspergillus nidulans. Previously we have reported the molecular characterization of a number of in vivo selected mutant alleles and showed that they were unlikely to represent total loss of function alleles (Shroff et al., 1996) and that a deletion of the creA gene and surrounding DNA has an extremely severe effect on morphology under both carbon catabolite repressing and carbon catabolite nonrepressing conditions (Dowzer and Kelly, 1991). Here we present an analysis of in vivo selected creA mutations with an extreme morphological phenotype and show that some of these alleles would be predicted to result in no functional CreA. The most extreme of these alleles resulted in a truncation of the protein within the first zinc finger. Precise gene disruptions, leaving the flanking sequences intact, show essentially the same phenotype as this truncated allele. Thus, a strain containing a null allele is viable, and the leaky-lethal phenotype of previous deletion alleles (Dowzer and Kelly, 1991) must be due to the deletion of additional 3' genomic sequence. A strain containing an allele that results in a deletion of the final 80 amino acids shows reduced sensitivity to carbon catabolite repression for a number of systems, thus localizing a region of the protein involved in repression. Surprisingly, the phenotypically most extreme allele studied is not a null allele, but results in an amino acid substitution that would disrupt the zinc finger region and abolish binding to DNA. This is the only allele that produces a full-length protein, predicted to be nuclear localized, but which completely abolishes DNA binding. The phenotype may be more extreme than the null alleles due to the nuclear located CreA protein titrating interacting proteins. PMID- 9344630 TI - Cerato-ulmin, a hydrophobin secreted by the causal agents of Dutch elm disease, is a parasitic fitness factor. AB - Dutch elm disease is caused by the aggressive Ophiostoma novo-ulmi and the nonaggressive O. ulmi. Both secrete the protein cerato-ulmin (CU). To determine what role CU plays in the pathology of Dutch elm disease, we constructed a CU overexpression mutant of the nonaggressive O. ulmi H5. Stable integration of a single copy of the cu gene from the aggressive O. novo-ulmi into the genome of the nonaggressive isolate resulted in increased secretion of CU protein. Trials with American elm, Ulmus americana, suggested no alteration of virulence of this overexpressing transformant. Using aggressive and nonaggressive wild types, the cu overexpressing mutant, and our cu- mutant (Bowden et al., 1996), we have demonstrated that CU production is correlated with an altered phenotype and more hydrophobic and adherent yeast-like cells. Our results also demonstrate that CU has a role in protecting infectious propagules from desiccation. These biological roles for CU would affect transmission of Dutch elm disease, and we therefore propose that this hydrophobin acts as a parasitic fitness factor. PMID- 9344631 TI - Expression of two closely linked hydrophobin genes of Coprinus cinereus is monokaryon-specific and down-regulated by the oid-1 mutation. AB - A protein with characteristic properties of a fungal hydrophobin (CoH1) was isolated from the monokaryotic stage of the basidiomycete Coprinus cinereus. A cosmid clone containing the corresponding gene (coH1) was identified using a cDNA probe derived by RT-PCR. Hybridization and sequence analysis identified a second gene, coH2, just 4.1 kb downstream of coH1 encoding a hydrophobin (CoH2) with 64% sequence identity. Both coH1 and coH2 are subject to developmental regulation. They are expressed in vegetative monokaryotic cells but not in the asexual oidia produced on the surface of monokaryons. Transcripts of the genes were barely detected in dikaryotic mycelium and were absent from fruit bodies. Loss of aerial growth due to a mutation known as oid-1 was correlated with lack of both hydrophobins. PMID- 9344632 TI - Expression of angiotensin type-1 (AT1) and type-2 (AT2) receptor mRNAs in the adult rat brain: a functional neuroanatomical review. AB - The discovery that all components of the renin-angiotensin system (RAS) are present in the central nervous system led investigators to postulate the existence of a local brain RAS. Supporting this, angiotensin immunoreactive neurons have been visualized in the brain. Two major pathways were described: a forebrain pathway which connects circumventricular organs to the median preoptic nucleus, paraventricular nucleus, and supraoptic nucleus, and a second pathway connecting the hypothalamus to the medulla oblongata. Blood-brain barrier deficient circumventricular organs are rich in angiotensin II receptors. By activating these receptors, circulating angiotensin II may act on central cardiovascular centers via angiotensinergic neurons, providing a link between peripheral and central angiotensin II systems. Among the effector peptides of the brain RAS, angiotensin II and angiotensin III have the same affinity for the two pharmacologically well-defined receptors: type 1 (AT1) and type 2 (AT2). When injected in the brain, these peptides increase blood pressure, water intake, and anterior and posterior pituitary hormone release and may modify memory and learning. The cloning of AT1 and AT2 receptor cDNAs has revealed that these receptors belong to the seven transmembrane domain receptor family. In rodents, two AT1 receptor subtypes, AT1A and AT1B, have been isolated. Using specific riboprobes for in situ hybridization histochemistry, recent studies mapped the distribution of AT1A, AT1B, and AT2 receptor mRNAs in the adult rat and found a predominant expression of AT1A and AT2 mRNA in the brain and of AT1B in the pituitary. Very limited overlap was found between the brain expression of AT1A and AT2 mRNAs. In several functional entities of the brain, such as the preoptic region, the hypothalamus, the olivocerebellary system, and the brainstem baroreflex arc, the colocalization of receptor mRNA, binding sites, and angiotensin immunoreactive nerve terminals suggests local synthesis and expression of angiotensin II receptors. In other areas, such as the bed nucleus of the stria terminalis, the median eminence, or certain parts of the nucleus of the solitary tract, angiotensin II receptors are likely of extrinsic origin. The neuronal expression of AT1A and AT2 receptors was demonstrated in the subfornical organ, the hypothalamus, and the lateral septum. By using double label in situ hybridization, AT1A receptor expression was localized in corticotropin releasing hormone but not in vasopressin containing neurons in the hypothalamus. The information is discussed together with functional data concerning the role of brain angiotensins, in an attempt to provide a better understanding of the physiological and functional roles of each receptor subtype. PMID- 9344633 TI - Central hypertensive effects of aldosterone. AB - The soluble mineralocorticoid receptor bound to an agonist acts as a transcription factor for several genes relevant to ion transport by kidney and colon epithelial cells and is a major regulator of electrolyte and fluid homeostasis. Mineralocorticoids, the most prominent of which is aldosterone, also influence the activity of nonepithelial target cells, including vascular smooth muscle cells, by altering intracellular ion transport and content. Evidence is summarized for mineralocorticoid modulation of neuronal activity in a center or centers within the brain, probably in the periventricular area of the anterior hypothalamus, where information on electrolyte, fluid, and cardiovascular status is received and integrated, resulting in alterations in central sympathetic efferent activity. These functions are distinct from central aldosterone effects on salt appetite and peripheral trophic effects on cardiovascular tissue. The isolated mineralocorticoid receptor binds several adrenal steroids, including aldosterone and the major glucocorticoids, with equal affinity. Ligand specificity for the mineralocorticoid receptor differs between tissues, including different organs in the brain. Specificity is conferred extrinsically by the 11 beta-hydroxysteroid dehydrogenase enzymes in transport epithelia, but mechanisms for mineralocorticoid ligand specificity have not been completely defined in the brain. The functional interaction between the mineralocorticoid receptor bound to different ligands and between the mineralocorticoid and glucocorticoid receptors is complex and as yet unresolved. Evidence is presented for the de novo synthesis of adrenal corticosteroids in the brain which may, by paracrine regulation of central control mechanisms, be relevant for certain clinical and experimental forms of hypertension characterized by low circulating levels of mineralocorticoids which respond to mineralocorticoid receptor antagonists. PMID- 9344634 TI - Effects of nitric oxide on neuroendocrine function and behavior. AB - Nitric oxide (NO) is an unusual chemical messenger. NO mediates blood vessel relaxation when produced by endothelial cells. When produced by macrophages, NO contributes to the cytotoxic function of these immune cells. NO also functions as a neurotransmitter and neuromodulator in the central and peripheral nervous systems. The effects on blood vessel tone and neuronal function form the basis for an important role of NO on neuroendocrine function and behavior. NO mediates hypothalamic portal blood flow and, thus, affects oxytocin and vasopression secretion; furthermore, NO mediates neuroendocrine function in the hypothalamic pituitary-gonadal and hypothalamic-pituitary-adrenal axes. NO influences several motivated behaviors including sexual, aggressive, and ingestive behaviors. Learning and memory are also influenced by NO. Taken together, NO is emerging as an important chemical mediator of neuroendocrine function and behavior. PMID- 9344635 TI - Vigabatrin versus carbamazepine and phenytoin in kainic acid-treated pubescent rats. AB - The effects of the administration of two doses (1,000 and 1,500 mg kg-1) of gamma vinyl-GABA (GVG), have been tested in pubescent rats, systemically injected with kainic acid (KA). The changes in spontaneous behaviour before KA injection, the behavioural and epileptic manifestations (Wet Dog Shakes, Limbic Seizures and Status Epilepticus) and the lethality rate caused by KA were taken into account and compared to those observed in controls and in carbamazepine (CBZ) or phenytoin (PHT) treated animals. While GVG appeared to reduce the incidence of the epileptic manifestations and the subsequent mortality, particularly when higher doses of the drug were used, CBZ exerted a proconvulsant action and PHT did not substantially modify the parameters considered. Moreover, GVG, but not CBZ and PHT, induced remarkable sedative effects which disappeared within 48 h. The different anticonvulsant profile of GVG, CBZ and PHT were correlated to their different modes of action, since GVG acts by enhancing the inhibitory GABA mediated processes, while CBZ and PHT act by reducing the excitatory processes. PMID- 9344636 TI - Effect of amiloride A Na+/H+ exchange inhibitor on cardioprotective effect of ischaemic preconditioning: possible involvement of resident cardiac mast cells. AB - The present study was designed to investigate the effect of amiloride, a Na+/H+ exchange inhibitor on cardioprotective effect of ischaemic preconditioning in isolated rat heart. Four episodes of ischaemic preconditioning and amiloride (174 microM) treatment markedly decreased the release of lactate dehydrogenase (LDH) and creatine kinase (CK) in coronary effluent and infarct size in hearts subjected to 30 min of global ischaemia followed by 120 min of reperfusion. Amiloride (174 microM) administered during ischaemic preconditioning (Amiloride in Ischaemic Preconditioning), produced no marked effect on LDH and CK release and infarct size. Ischaemic preconditioning and amiloride treatment significantly reduced ischaemia/reperfusion induced release of mast cell peroxidase (MPO). Four episodes of pH (20 mm of ammonium chloride) preconditioning also produced cardioprotection and decreased ischaemia/reperfusion induced release of MPO. It is interesting to note that a significant increase in release of MPO was observed immediately after ischaemic preconditioning and the release was inhibited by amiloride. Moreover, similar increase in MPO release was noted immediately after pH preconditioning. These findings tentatively suggest that ischaemic preconditioning and pH preconditioning produced cardioprotective effect by activating Na+/H+ exchange and consequent degranulation of resident cardiac mast cells. Amiloride administered during ischaemic preconditioning attenuated the cardioprotective effect of ischaemic preconditioning. PMID- 9344637 TI - Effects of supplementation with iloprost on the cardioprotection by BW755C (a dual inhibitor of cyclooxygenase and lipoxygenase enzymes) in myocardial reperfusion injury. AB - The effect of BW755C (a dual inhibitor of cyclo- and lipoxygenase enzymes) alone and in combination with iloprost (a PGI2 analogue) on myocardial reperfusion injury was investigated in anaesthetised open-chest dogs. The left anterior descending coronary artery was occluded for a period of 40 min followed by reperfusion for 3 h. Dogs were administered either saline, BW755C (10 mg kg-1 slow bolus) or BW755C plus iloprost (100 ng kg-1 min-1 for 75 min) just prior to reperfusion. The haemodynamic data showed significant reduction in MAP and both LV peak-positive and peak-negative dP/dt following reperfusion in the saline treated group, along with a delayed recovery of LVEDP. These changes were accompanied by significant depletion of myocardial ATP and glycogen stores. Administration of BW755C prevented the haemodynamic changes and replenished the HEP stores. Although coadministration of iloprost with BW755C afforded early normalisation of LVEDP and LV peak positive dP/dt, but MAP and LV peak negative dP/dt remained significantly depressed. Therefore, it might be concluded from this study that supplementation of BW755C with iloprost may have deleterious haemodynamic effects on the reperfused myocardium, particularly in the doses used. PMID- 9344638 TI - Attenuation in rat brain nitric oxide synthase activity in the coarctation model of hypertension. AB - The correlation of brain nitric oxide synthase (NOS) activity with renal hypertension has been investigated in rats. NOS activity was measured by oxyhaemoglobin and by conversion of radioactive arginine to citrulline. There was significant elevation of blood pressure (54% increase) along with left ventricular hypertrophy (26% greater than the control) 8 weeks after coarctation. The brain NOS activity was also significantly reduced in coarctated animals (45% of the control value). Treatment with captopril (angiotensin converting enzyme inhibitor) or centhaquin (centrally acting antihypertensive agent) led to significant reduction of left ventricular hypertrophy and a marked recovery in the brain NOS activity (to 92% and 135% of the control, respectively). Nifedipine (a calcium channel blocker) also brought about normalization of blood pressure but the left ventricular hypertrophy was not prevented. The brain NOS activity in the nifedipine treated group also showed a significant trend of recovery (to 73% of the control NOS activity). The results suggested that there is an inverse correlation between brain NOS activity and blood pressure level. PMID- 9344639 TI - Proapoptotic effect of atorvastatin on stimulated rabbit smooth muscle cells. AB - The in vitro and in vivo activity of atorvastatin and other 3-hydroxy-3 methylglutaryl coenzyme A (HMGCoA) reductase inhibitors (fluvastatin, pravastatin and simvastatin) has been investigated. Atorvastatin, fluvastatin, pravastatin and simvastatin caused a significant and dose-dependent (0.1-50 microM) reduction in cell multiplication of vascular smooth muscle cells (SMC) in cultures associated with the retardation of cycling cells in the G1 and G2/M compartments at 24 h, a phenomenon leading to apoptosis (programmed cell death) in several experimental in vitro models. The effects on the cell cycle resulted in a strong inhibition of cell proliferation at 48 h, followed by apoptosis when incubation was prolonged to 72 h as assessed by nuclei morphology and cytofluorimetric analysis of DNA. The apoptotic effect observed for the statins is completely prevented by the addition of exogenous mevalonate at a 100 microm concentration. in vivo SMC proliferation was stimulated by applying a silastic collar to the outside surface of carotid arteries in normocholesterolemic rabbits in the presence of an anatomically intact endothelium. The positioning of the collar promoted apoptosis in control vessels as assessed by Terminal Deoxynucleotidyl Transferase-dUTP-Biotin Nick-End Labeling (TUNEL) assay. The pre-treatment with 5 mg kg-1 per day of atorvastatin before collar insertion strongly increased the number of TUNEL-positive cells, suggesting a pro-apoptotic effect of HMGCoA reductase inhibitors also in vivo, even though cell DNA rearrangement still needs to be excluded. No apoptotic signal was detectable in sham operated arteries with no collar in either control or atorvastatin-treated rabbits. These data indicate that HMGCoA reductase inhibitors effect on the arterial wall may involve the modulation of both cell proliferation and programmed cell deaths supporting a possible role of statins in the prevention of early lesion and restenosis. PMID- 9344640 TI - Microcirculatory disturbances of the pancreas in cerulein-induced acute pancreatitis in rats with reference to L-arginine, heparin, and procaine treatment. AB - Local microcirculatory dysfunction within the pancreatic gland might be an important factor in the conversion of oedematous to necrotizing pancreatitis. Therapeutic agents, improving the pancreatic blood flow, might be valuable in acute pancreatitis treatment. An influence of nitric oxide, heparin and procaine treatment on microcirculatory values in acute pancreatitis (AP) in rats was investigated. Acute pancreatitis was induced by i.p. injection of cerulein in four doses of 15 microg kg-1 each at 1-h intervals. The rats with pancreatitis were divided into five groups, 12 animals each. One group remained without treatment, four groups were treated i.p. either with NO synthase inhibitor L-NNA (2x25 mg kg-1 or heparin 2x2.5 mg kg-1 or L-arginine 2x100 mg kg-1 or procaine 2x25 mg kg-1. Five control groups, ten animals each, received saline, L-NNA, heparin, L-arginine or procaine only. Five hours after the first ceruleine injection microcirculatory values within the pancreas were measured by means of laser Doppler flowmetry. Acute pancreatitis caused a significant drop of microcirculatory value to 37% of the basal value. The L-NNA administration resulted in a further insignificant reduction of the pancreatic blood flow to 34%. An improvement of microcirculation was observed in rats with pancreatitis receiving heparin (76%) and L-arginine (72%). Procaine had no effect on microcirculatory disturbances within the pancreas in rats with pancreatitis. Cn induced acute pancreatitis (AP) causes microcirculatory deterioration within the pancreas. Heparin and nitric oxide donor, L-arginine, might be considered as therapeutic agents, improving the diminished pancreatic tissue perfusion observed in acute pancreatitis. Procaine does not improve the pancreatic blood flow in acute pancreatitis. PMID- 9344642 TI - High rate of intestinal absorption of the phospholipid anlaogue 1-dodecyl 2-[1 14C] octanamido-sn-2-deoxy-glycero-3-phosphocholine in the rat. AB - The phospholipid analogue with two short fatty chains, 1-dodecyl-2-[1-14C] octanamido-sn-2-deoxy-glycero-3-phosphocholine ([14C] phospholipid analogue), with a non-hydrolyzable bond at position 2 of the glycerol, is an inhibitor of phospholipase A2. It was obtained after chemical synthesis and 0.5 micromol was solubilized in Na+ taurocholate with an equimolar amount of 1-octadecanoyl 2 [3H]eicosatetraenoyl-sn- glycero-3-phosphocholine which is the current substrate of phospholipases A2. Both molecules were introduced into the duodenum of rats in order to follow their captations by intestinal mucosa cells for 30, 60 or 90 min. The [14C] phospholipid analogue was poorly split by phospholipases A2 (pancreatic juice and intracellular enzymes). It disappeared from the intestinal contents (87% of the dose gone in 90 min) as rapidly as the tritiated lecithin (81%) but this was later split by the phospholipases at a higher rate. PMID- 9344641 TI - Glibenclamide sensitivity of neural relaxation of the rabbit sphincter of oddi. AB - In this article we studied whether the nitrergic relaxation of the rabbit sphincter was sensitive to glibenclamide. Field stimulation relaxed the sphincter of Oddi rings after incubation with atropine (1 microM) and guanethidine (4 microM) with threefold and fourfold increases in tissue guanosine 3':5'-cyclic monophosphate and adenosine 3':5'-cyclic monophosphate contents, respectively. These changes were blocked by 30 microM NG-nitro-L-arginine methyl ester. Glibenclamide (0.1-10 microM) attenuated the field stimulation-induced relaxation and completely abolished the relaxation produced by 0.1 microM cromakalim. We conclude that nitrergic relaxation of the rabbit sphincter of Oddi comprises a mechanism sensitive to glibenclamide. PMID- 9344644 TI - Contractile effects of porcine galanin(1-29)-NH2 on the rat isolated gastric fundus: mediation by potassium ions. AB - Galanin (3-300 nM) evoked reproducible concentration-dependent contractions of rat isolated gastric fundus strips, EC50 of the peptide equalled 16.7 nM (6.2 39.2) and the slope of the concentration-response curve was 34.8 (24.0-45.7). The maximal response (Emax) to carbachol (30 nM) was not affected by the absence of the potassium ions in the bathing solution. On the contrary the Emax to galanin (300 nM) was decreased by almost 95% by the use of the potassium-free buffer. Re exposure of the muscle strips to potassium containing bathing medium reversed the inhibition by about 35%, yet the value remained significantly lower than that of the control. Apamin (1 and 2 microM), glybenclamide (10 microM), clofilium tosylate (10 microM) did not significantly influence the Emax to carbachol. Apamin or glybenclamide did not affect the contractile action of galanin, while clofilium attenuated the Emax to the peptide in a concentration-dependent manner, the EC50 of the agent being 9.44 microM (164 nM-541 microM). It was concluded that the potassium ions play a modulatory role in gastric smooth muscle contraction following galanin receptor stimulation, probably by interacting with the extracellular calcium influx. PMID- 9344643 TI - Effect of cytochalasin D on systemic and local anaphylaxis in a murine model. AB - We investigated the effect of cytochalasin D on anaphylaxis. Cytochalasin D dose dependently inhibited systemic anaphylaxis induced by compound 48/80 in rats. Especially, cytochalasin D inhibited compound 48/80-induced systemic anaphylaxis 100% with a dose of 1 microg g-1 body weight (BW). Cytochalasin D significantly inhibited serum histamine levels induced by compound 48/80. Cytochalasin D (10( 1) microg g-1 BW) also inhibited local anaphylaxis activated by anti dinitrophenyl (DNP) IgE to 79.6+/-1.8%. Cytochalasin D dose-dependently inhibited histamine release from the rat peritoneal mast cells activated by compound 48/80 or anti-DNP IgE. The level of cAMP in rat peritoneal mast cells, when cytochalasin D was added, transiently and significantly increased about fourfold compared with that of basal cells. Our studies provide evidence that cytochalasin D will be beneficial in the treatment of anaphylaxis. PMID- 9344645 TI - Tachykinins as peripheral modulators of primary afferent nerves and visceral sensitivity. PMID- 9344646 TI - Positional cloning of novel skin-specific genes from the human epidermal differentiation complex. AB - The epidermal differentiation complex, located on human chromosomal band 1q21, contains at least 20 genes expressed during epidermal differentiation. We constructed a 1.2-Mb YAC contig spanning the SPRR and S100 gene clusters. Restriction mapping and FISH confirmed the colinearity of the contig with the genomic restriction map (A. Volz et al., 1993, Genomics 18:92-99). However, the YAC clones revealed several additional restriction sites not previously detected in genomic DNA, presumably due to CpG methylation. Making use of cDNA selection, we have identified three novel cDNAs, all of which map to the SPRR/IVL region. All three transcripts are expressed at high levels in normal and psoriatic skin, but not in cultured keratinocytes or in a variety of cell lines and human tissues. The molecular cloning of this region provides a valuable tool for identifying additional epidermal differentiation genes and for elucidating the relationship between chromatin structure and gene expression during terminal differentiation. PMID- 9344647 TI - ARVD4, a new locus for arrhythmogenic right ventricular cardiomyopathy, maps to chromosome 2 long arm. AB - Autosomal dominant arrhythmogenic right ventricular dysplasia (ARVD; MIM 107970) is a genetically heterogeneous cardiomyopathy, which often causes sudden death in juveniles and athletes. Two disease loci were previously mapped respectively to 14q23-q24 (ARVD1) and to 1q42-q43 (ARVD2). A third possible locus was assigned to 14q12-q22. We report here on a linkage study performed on three independent families with recurrence of ARVD characterized by localized involvement of the left ventricle. In these families the disease appears to be transmitted with three polymorphic DNA markers of the chromosome 2 long arm, showing a maximum lod score of 3.46 at theta = 0 for the marker D2S152. The multipoint linkage analysis suggests that the novel ARVD locus, provisionally named ARVD4, maps to 2q32. 1 q32.3, within the chromosomal region including markers D2S152, D2S103, and D2S389. PMID- 9344648 TI - Fine structure of the murine leptin receptor gene: splice site suppression is required to form two alternatively spliced transcripts. AB - The fine structure of the murine leptin receptor gene (Lepr) is described. Duplicated ligand binding domains (conserved among cytokine receptors) are found in eight exons (coding exons 3 to 6 and 8 to 11). Thus, it is possible that a single leptin receptor molecule could have two functional ligand binding domains. The transmembrane region of Lepr is in coding exon 16 while the juxtamembrane JAK docking site is in coding exon 17. For all membrane-bound forms, the transcript must include 17 invariant exons and 1 alternatively spliced 3' terminal exon. The transcript encoding the soluble receptor (Re) includes 14 coding exons and an alternatively spliced 3' terminal exon. We have identified two splice variants (Rc and Re) for which there are no intervening sequences between the two final exons. This unusual juxtaposition of exons requires that splice donor sites at the 5' end of the respective terminal exons be ignored in the production of these splice variants. We suggest that splice site suppression is responsible for the formation of two of the alternatively spliced forms of the mouse Lepr gene. The juxtaposition of two coding exons separated by a consensus splice donor sequence is the structural substrate for this mode of alternative splicing. We present evidence that the Rc form is expressed in human tissues while the Re form, the soluble receptor, is not expressed. PMID- 9344649 TI - A sequence-ready physical map of a region of 12q24.1. AB - We developed a sequence-ready map of a part of human chromosome 12q24.1. We utilized a number of sequence-tagged site (STS) markers from 12q24.1 to screen large insert bacterial chromosome libraries and a chromosome 12-specific cosmid library. The clones were assembled into contiguous sets (contigs) by STS-content analysis. Contigs were extended by obtaining end sequences of bacterial clones, generation of additional STSs, rescreening the libraries, and screening the additional clones for the presence of STSs. The resulting contig covers nearly 2 Mb of DNA and provides an average marker resolution of 16 kb. Based on the STS content, we developed fingerprints of a subset of clones. The STS content and fingerprint data allowed us to define a minimal tiling path of clones. These clones are being used to sequence this part of chromosome 12. This contig contains the Ataxin 2 gene, and it covers the interval harboring the gene responsible for Darier disease. PMID- 9344650 TI - Identification of putative transmembrane receptor sequences homologous to the calcium-sensing G-protein-coupled receptor. AB - The sensing of extracellular calcium is a general paradigm for regulating diverse cellular functions in many tissues. A calcium-sensing receptor (Casr) belonging to the metabotropic glutamate family of G-protein-coupled receptors (GPCR) that transduces the effects of extracellular calcium in the parathyroid gland as well as other tissues has been identified. The diversity of GPCR families and the recent finding of calcium sensing in cells lacking the known Casr suggest the existence of additional receptors related to Casr. By polymerase chain reaction (PCR) amplification and screening of genomic libraries, we have identified multiple Casr-related sequences (Casr-rs) in the mouse. Using primers designed to regions of the first and third intracellular loops of Casr, we initially PCR amplified a 497-bp Casr-related sequence (Casr-rs1) with high homology to Casr. The deduced protein sequence of Casr-rs1 is 63% similar and 40% identical to Casr over the available transmembrane region. We screened a mouse genomic library with a Casr-rs1 probe and identified two additional Casr-related sequences (Casr-rs2 and Casr-rs3). In the predicted transmembrane domain, Casr-rs2 and Casr-rs3 are 95% identical to Casr-rs1. We mapped Casr-rs1 to mouse Chromosome (Chr) 7 by interspecific backcross analysis, whereas the known Casr localizes to mouse Chr 16. By fluorescence in situ hybridization, Casr-rs2 also localized to mouse Chr 7 and Casr-rs3 mapped to mouse Chr 4. We were able to distinquish Casr-rs1 from Casr-rs2 by PCR using specific primers, suggesting that they are distinct genes clustered on Chr 7. By RT-PCR, we identified additional Casr-rs transcripts in mouse kidney, brain, testis, embryo, and MC3T3-E1 osteoblasts, but not in lung or liver. The homologous sequence in mouse kidney, embryo, and MC3T3-E1 osteoblasts, designated Casr-rs4, has a deduced amino acid sequence that is 100% similar and 97% identical to that of Casr-rs1. The sequence amplified from mouse brain, Casr rs5, has a deduced protein sequence that is 96% similar and 92% identical to that of Casr-rs1. Our findings establish the existence of a novel multimembered family of Casr-related sequences in the mouse which may encode receptors that transduce responses to diverse extracellular cations. PMID- 9344651 TI - Structure and expression of the mouse L23mrp gene downstream of the imprinted H19 gene: biallelic expression and lack of interaction with the H19 enhancers. AB - The human L23 (mitochondrial)-related protein gene, located 40 kb downstream of the imprinted H19 gene, is biallelically expressed. We have cloned and characterized its mouse homolog, L23mrp, which maps to the conserved syntenic region on mouse chromosome 7. The promoter of L23mrp is a CpG island that is transcribed ubiquitously, but at different levels, in different fetal tissues. Allele-specific expression analysis revealed that both parental alleles are equally active. Since the enhancers located between H19 and L23mrp had been shown to be involved in the imprinted expression of Ins-2, Igf-2, and H19, we asked whether they also influence L23mrp. Analysis of mice with a targeted deletion of the enhancers demonstrated that they were not disrupted in the expression of L23mrp. These findings indicate that L23mrp is functionally insulated from the Ins-2/Igf-2/H19 domain in terms of both imprinting and enhancer action. PMID- 9344652 TI - Chromosome localization and structure of the murine cyclin G1 gene promoter sequence. AB - Cyclins play an essential role in the control of the cell cycle. In this study the murine cyclin G1 gene expression, structure, and chromosomal localization were examined. Genes with high homology to murine cyclin G1 were detected in various mammals, including human, monkey, rat, dog, cow, and rabbit, but not in yeast or chicken. Cyclin G1 gene was expressed in all murine tissues examined, with the highest levels in cardiac and skeletal muscle. A 10,366-bp genomic DNA fragment encompassing the promoter region and the 5'-flanking region of the gene was cloned and sequenced. Three putative binding sites for the myocyte enhancer factor-2 family of transcription factors were revealed. Furthermore, an upstream p53-binding site was localized to nucleotides -252 to -233 and a new putative p53 binding site was identified in the first intronic region at nucleotides 275 to 294. By fluorescence in situ hybridization, the cyclin G1 gene was mapped to mouse chromosome 11B1.1. This region is homologous with human chromosome 5q31 q32, consistent with the recent mapping of the human cyclin G1 gene to chromosome 5q32-q34. Localization of murine cyclin G1 will facilitate determination of gene linkage and the identification of synteny groups in mammals and of DNA elements in or near this gene that mediate its tissue expression or development-specific pattern of expression. PMID- 9344653 TI - Structure of the human type XIX collagen (COL19A1) gene, which suggests it has arisen from an ancestor gene of the FACIT family. AB - Type XIX collagen is a newly discovered member of the FACIT (fibril-associated collagens with interrupted triple helices) group of extracellular matrix proteins. Based on the primary structure, type XIX collagen is thought to act as a cross-bridge between fibrils and other extracellular matrix molecules. Here we describe the complete exon/intron organization of COL19A1 and show that it contains 51 exons, spanning more than 250 kb of genomic DNA. The comparison of exon structures of COL19A1 and other FACIT family genes revealed several similarities among these genes. The structure of exons encoding the noncollagenous (NC) 1-collagenous (COL) 1-NC 2-COL 2-NC 3-COL 3-NC 4 domain of the alpha1(XIX) chain is similar to that of the NC 1-COL 1-NC 2-COL 3-NC 3 domain of the alpha2(IX) chain except for the NC 3 domain of alpha1(XIX). The exons encoding the COL 5-NC 6 domain of alpha1(XIX) are also similar to those of the COL 3-NC 4 domain of alpha1(IX) chain. Previously, COL19A1 was mapped to human chromosome 6q12-q14, where COL9A1 is also located. Likewise, the present work shows that the mouse Col19a1 gene is located on mouse chromosome 1, region A3, where Col9a1 has also been mapped. Taken together, the data suggest that COL19A1 and COL9A1 (Col19a1 and Col9a1) were duplicated from the same ancestor gene of the FACIT family. Three CA repeat markers with high heterozygosity were found in COL19A1. These markers may be useful for linkage analysis of age-related inheritable diseases involved in eyes and/or brain. PMID- 9344654 TI - Mouse chromosomal locations of nine genes encoding homologs of human paraneoplastic neurologic disorder antigens. AB - The paraneoplastic neurologic disorders (PND) are a rare group of neurologic syndromes that arise when an immune response to systemic tumors expressing neuronal proteins ("onconeural antigens") develops into an autoimmune neuronal degeneration. The use of patient antisera to clone the genes encoding PND antigens has led to new insight into the mechanism of these autoimmune disorders. The tumor antigens can now be grouped into three classes: (1) neuron-specific RNA binding proteins, (2) nerve terminal vesicle-associated proteins, and (3) cytoplasmic signaling proteins. To understand better the evolutionary relatedness of these genes and to evaluate them as candidates for inherited neurological disorders, we have determined the mouse chromosomal locations of nine of these genes-Hua, Hub, Huc, Hud, Nova1, Nova2, Natpb, Cdr2, and Cdr3. These data suggest that the Hua-Hud genes arose from gene duplication and dispersion, while the other genes are dispersed in the genome. We also predict the chromosomal locations of these genes in human and discuss the potential of these genes as candidates for uncloned mouse and human mutations. PMID- 9344655 TI - Chromosomal mapping of the human and murine orphan receptors ERRalpha (ESRRA) and ERRbeta (ESRRB) and identification of a novel human ERRalpha-related pseudogene. AB - The estrogen-related receptors ERRalpha and ERRbeta (formerly ERR1 and ERR2) form a subgroup of the steroid/thyroid/retinoid receptor family. ERRalpha and ERRbeta are homologous to the estrogen receptor and bind similar DNA targets; however, they are unable to activate gene transcription in response to estrogens. We have used interspecific backcross analysis to map the murine Estrra locus to chromosome 19 and Estrrb to mouse chromosome 12. Using fluorescence in situ hybridization, we have mapped the human ESRRA gene to chromosome 11q12-q13 and the human ESRRB gene to chromosome 14q24.3. In addition, we report the isolation of a processed human ERRalpha pseudogene mapping to chromosome 13q12.1. To our knowledge, this represents the first report of a pseudogene associated with a member of the nuclear receptor superfamily. PMID- 9344656 TI - Identification of two novel human putative serine/threonine kinases, VRK1 and VRK2, with structural similarity to vaccinia virus B1R kinase. AB - A cDNA library enriched for human fetal-specific liver genes was constructed by suppressive subtractive hybridization. EST fls223 generated from this library was found to represent a novel putative serine/threonine (Ser/Thr) kinase. A full length clone isolated for this gene encodes a protein of 396 amino acids. The amino acid sequence has 40% identity over 305 amino acids with the B1R Ser/Thr protein kinase of vaccinia virus. This gene has therefore been named VRK1 (vaccinia virus B1R kinase related kinase). VRK1 was also found to have sequence identity (62.0% over 481 nucleotides) to a database EST. A full-length clone for this EST was isolated and sequenced. Conceptual translation predicts a protein of 508 amino acids that, like VRK1, has similarity to B1R kinase (38.7% identity over 300 amino acids). This gene has been named VRK2. Comparison of VRK1 with VRK2 indicates that they encode structurally related putative Ser/Thr protein kinases. Northern analysis shows that expression of both genes is widespread and elevated in highly proliferative cells, such as testis, thymus, and fetal liver. B1R kinase is reported to be essential for DNA replication of vaccinia virus. The similarity of VRK1 and VRK2 to B1R indicates that these genes may have similar functions. PMID- 9344657 TI - Genetic structure and chromosomal mapping of MyD88. AB - The myeloid differentiation (MyD) marker MyD88 was initially characterized as a primary response gene, upregulated in mouse M1 myeloleukemic cells in response to differentiation induced by interleukin-6. Subsequent analysis revealed that MyD88 possesses a unique modular structure, which consists of an N-terminal "death domain," similar to the intracellular segments of TNF receptor 1 and Fas, and a C terminal region related to the cytoplasmic domains of the Drosophila morphogen Toll and vertebrate interleukin-1 receptors. In this report we describe the cloning and gene structure of mouse MyD88. The complete coding sequence of mouse MyD88 spans five exons, with the first exon encoding the complete death domain. Zooblot analysis revealed that MyD88 is an evolutionarily conserved gene. MyD88 was localized to the distal region of mouse chromosome 9 by interspecific backcross mapping. The human homolog (hMyD88) was mapped to chromosome 3p22-p21.3 by PCR analysis of a human chromosome 3 somatic cell hybrid mapping panel. Northern blot analysis revealed widespread expression of MyD88 in many adult mouse tissues, and RT-PCR studies detected MyD88 mRNA in T and B cell lines and differentiating embryonic stem cells. The broad expression pattern demonstrates that mouse MyD88 expression is not restricted to cells of myeloid lineage as was originally believed. PMID- 9344658 TI - Sequence-based exon prediction around the synaptophysin locus reveals a gene-rich area containing novel genes in human proximal Xp. AB - The human Xp11.23-p11.22 interval has been implicated in several inherited diseases including Wiskott-Aldrich syndrome; three forms of X-linked hypercalciuric nephrolithiaisis; and the eye disorders retinitis pigmentosa 2, congenital stationary night blindness, and Aland Island eye disease. In constructing YAC contigs spanning Xp11. 23-p11.22, we have previously shown that the region around the synaptophysin (SYP) gene is refractory to cloning in YACs, but highly stable in cosmids. Preliminary analysis of the latter suggested that this might reflect a high density of coding sequences and we therefore undertook the complete sequencing of a SYP-containing cosmid. Sequence data were extensively analyzed using computer programs such as CENSOR (to mask repeats), BLAST (for homology searches), and GRAIL and GENE-ID (to predict exons). This revealed the presence of 29 putative exons, organized into three genes, in addition to the 7 exons of the complete SYP coding region, all mapping within a 44-kb interval. Two genes are novel, one (CACNA1F) showing high homology to alpha1 subunits of calcium channels, the other (LMO6) encoding a product with significant similarity to LIM-domain proteins. RT-PCR and Northern blot studies confirmed that these loci are indeed transcribed. The third locus is the previously described, but not previously localized, A4 differentiation-dependent gene. Given that the intron-exon boundaries predicted by the analysis are consistent with previous information where available, we have been able to suggest the genomic organization of the novel genes with some confidence. The region has an elevated GC content (>53%), and we identified CpG islands associated with the 5' ends of SYP, A4, and LMO6. The order of loci was Xpter-A4 LMO6-SYP-CACNA1F-Xcen, with intergenic distances ranging from approximately 300 bp to approximately 5 kb. The density of transcribed sequences in this area (>80%) is comparable to that found in the highly gene-rich chromosomal band Xq28. Further studies may aid our understanding of the long-range organization surrounding such gene-enriched regions. PMID- 9344659 TI - The uroguanylin gene (Guca1b) is linked to guanylin (Guca2) on mouse chromosome 4. AB - Uroguanylin is an endogenous ligand of the intestinal receptor guanylate cyclase C (GC-C). Both uroguanylin and the related peptide ligand guanylin bind to GC-C and stimulate an increase in cyclic GMP, inducing chloride secretion via the cystic fibrosis transmembrane conductance regulator. We describe the cloning of the complete mouse uroguanylin gene (Guca1b) and show that Guca1b is tightly linked to the mouse guanylin gene on chromosome 4. The two genes are structurally similar, being composed of three short exons; the uroguanylin gene spans 2.4 kb and the guanylin gene spans 1.7 kb. Uroguanylin mRNA is most prominent in proximal small intestine, whereas guanylin mRNA is predominantly expressed in distal small intestine and colon. The upstream promoter sequence of the mouse uroguanylin gene contains a canonical TATA element at the site of transcription initiation and consensus binding sites for several known transcription factors, including HNF-1 and Sp1 within the first 1 kb. Although the gene structure and coding sequences of uroguanylin and guanylin are similar, the 5' flanking sequences and patterns of expression of these two genes in the intestine are different. It is likely that uroguanylin and guanylin represent gene duplications that have evolved to allow overlapping and complementary patterns of expression in the intestine. PMID- 9344660 TI - Multiple functional copies of the RBM gene family, a spermatogenesis candidate on the human Y chromosome. AB - The RBM (RNA-binding motif) gene family on the human Y chromosome encodes proteins with an RNA-binding domain. Its exclusive expression in germ cells and its partial deletion in some azoospermic or severely oligospermic males provide evidence of a role for RBM genes in spermatogenesis. There are approximately 30 RBM genes, found on both arms of the Y chromosome. Two RBM cDNA clones with slightly different sequences have been reported. To investigate the number of functional genes, we studied RBM expression by use of RT-PCR of RBM transcripts and by characterizing numerous RBM cDNA clones. A total of 27 RT-PCR and 19 cDNA clones were sequenced. Whereas the RT-PCR clones pointed to the existence of at least six RBM subfamilies (RBMI to RBMVI), the cDNA clones indicated that only RBMI is actively transcribed and encodes functional proteins. A total of six RBMI genes were identified, which produce four polypeptides due to some silent base substitutions. The transcripts of each gene are alternatively spliced to generate protein isoforms with three or four SRGY boxes, thus greatly increasing the complexity of the products of the RBM gene family. We also provide evidence suggesting that a 5-bp deletion in a previously reported RBM cDNA clone represents a processing irregularity. PMID- 9344661 TI - Genetic relationships of the genes encoding the human proteasome beta subunits and the proteasome PA28 complex. AB - Genomic clones were obtained for the genes encoding the beta subunits of the human proteasome and for the associated proteasome activators PA28alpha and beta (PSME1 and PSME2, respectively). Fluorescence in situ hybridization was used to map the gene encoding the beta subunit PSMB3 (beta3 hs, HsC10-II) to chromosome band 2q35, PSMB2 (beta4 hs, HsC7-I) to band 1p34.2, and PSMB4 (beta7 hs, HSBpros 26) to band 1q21. Genes encoding the alpha and beta subunits of the PA28 complex were found closely linked on chromosome band 14q11.2, within 1 Mb of the beta proteasome locus PSMB5 (beta5 hs, MB1, X). These data complete the mapping of the human proteasome beta subunit loci. With the exception of the genes encoding the PSMB9 and PSMB8 (LMP2 and LMP7, respectively) subunits, the beta genes were not closely linked in the human genome. Both PSMB2 and PSMB4 mapped to a region of chromosome 1 that is proposed to be paralogous to other regions of the human genome where beta proteasome genes map: chromosome 6 containing the major histocompatibility complex (MHC) and chromosome 9. The independent regulation of expression of all of these genes, implied by this study, is consistent with a key role for proteasome assembly in coordination of the complex. PMID- 9344663 TI - The structure and chromosome location of the human chondroadherin gene (CHAD). AB - The cDNA sequence of the human chondroadherin gene was cloned using PCR-based techniques. The gene encodes a protein of 359 amino acids, of which the first 21 amino acids represent a putative signal peptide sequence and which possesses 11 leucine-rich repeats flanked by cysteine-rich regions. The cDNA possesses a 5' untranslated region of 149 bp, a coding region of 1080 bp including the stop codon, and a 3' untranslated region of 561 bp terminating in a poly(A) tail. The cDNA hybridizes with a single messenger RNA of 1.9 kb, which is present in chondrocytes at all ages. Analysis of genomic DNA revealed that the chondroadherin gene possesses two introns, both of which reside within the coding region. The first intron has a length of about 2.3 kb and separates the codons for lysine(258) and phenylalanine(259). The second intron has a length of about 0.5 kb and splits the codon for tryptophan(314). This genomic organization results in exon 1 encoding the signal peptide, the amino-terminal cysteine-rich region, and the first 9 leucine-rich repeats; exon 2 encoding the last 2 leucine rich repeats and part of the carboxy-terminal cysteine-rich region; and exon 3 encoding the remainder of the carboxy-terminal cysteine-rich region. The gene does not possess a TATA box prior to its transcription start site. Isolation of a cosmid clone spanning the chondroadherin gene enabled its chromosome location to be established. The gene was shown to reside at chromosome 17q21.33. PMID- 9344662 TI - Analysis of the intron-exon organization of the human multidrug-resistance protein gene (MRP) and alternative splicing of its mRNA. AB - Overexpression of multidrug-resistance protein (MRP) and P-glycoprotein confers similar but not identical multidrug-resistance phenotypes. However, unlike P glycoprotein, which comprises two membrane-spanning domains (MSDs) and two nucleotide-binding domains, MRP contains a third NH2-proximal MSD, a feature now identified in several other ATP-binding cassette transmembrane transporters. MRP is located on chromosome 16 at band 13.1 close to the short-arm breakpoint of the pericentric inversion associated with the M4Eo subclass of acute myeloid leukemia. We have defined the intron-exon structure of MRP and characterized a number of splicing variants of MRP mRNA. The gene spans at least 200 kb. It contains 31 exons and a high proportion of class 0 introns, alternative splicing of which results in significant levels of variant transcripts that maintain the original open reading frame of MRP mRNA. Analyses of the conservation of intron exon organization and protein primary structure suggest that the MRP-related transporters evolved from a common ancestor shared with the cystic fibrosis transmembrane conductance regulator, by fusion with one or more genes encoding polytopic membrane proteins. PMID- 9344664 TI - Identification and characterization of the gene encoding a new member of the alpha-crystallin/small hsp family, closely linked to the alphaB-crystallin gene in a head-to-head manner. AB - alphaB-Crystallin is a member of the alpha-crystallin/small heat shock protein (hsp) family and under various neuropathologic conditions accumulates in reactive astrocytes and degenerating neurons. In the 5'-flanking region of the alphaB crystallin gene on human chromosome 11q22-q23, where a constitutive DNase I hypersensitive site is located, we identified a gene transcribed in the opposite direction. Analysis of its mRNA structure by RT-PCR and 5'/3'RACE revealed that this gene is composed of two exons and encodes a new member of the alpha crystallin/small hsp family. This gene was designated the HSPB2 gene by the HMGW Nomenclature Committee. The complete genomic structure of the rat homologue was also determined. Northern blot analysis revealed that the HSPB2 gene is expressed preferentially in skeletal muscle and heart but not in the lens, while the neighboring alphaB-crystallin gene is highly expressed in all three tissues. The two related genes are arranged in a head-to-head manner with an intergenic sequence of less than 1 kb, raising a possibility of shared regulatory elements for their expression. PMID- 9344665 TI - Human bZIP transcription factor gene NRL: structure, genomic sequence, and fine linkage mapping at 14q11.2 and negative mutation analysis in patients with retinal degeneration. AB - The NRL gene encodes an evolutionarily conserved basic motif-leucine zipper transcription factor that is implicated in regulating the expression of the photoreceptor-specific gene rhodopsin. NRL is expressed in postmitotic neuronal cells and in lens during embryonic development, but exhibits a retina-specific pattern of expression in the adult. To understand regulation of NRL expression and to investigate its possible involvement in retinopathies, we have determined the complete sequence of the human NRL gene, identified a polymorphic (CA)n repeat (identical to D14S64) within the NRL-containing cosmid, and refined its location by linkage analysis. Since a locus for autosomal recessive retinitis pigmentosa (arRP) has been linked to markers at 14q11 and since mutations in rhodopsin can lead to RP, we sequenced genomic PCR products of the NRL gene and of the rhodopsin-Nrl response element from a panel of patients representing independent families with inherited retinal degeneration. The analysis did not reveal any causative mutations in this group of patients. These investigations provide the basis for delineating the DNA sequence elements that regulate NRL expression in distinct neuronal cell types and should assist in the analysis of NRL as a candidate gene for inherited diseases/syndromes affecting visual function. PMID- 9344667 TI - Detailed physical analysis of a 1.5-megabase YAC contig containing the MXI1 and ADRA2A genes. AB - The distal long arm of chromosome 10 harbors genes of biomedical interest such as MXI1, a putative tumor suppressor gene, and those encoding the adrenergic receptors alpha2A (ADRA2A) and beta1 (ADRB1). As part of a physical and genetic study of this genomic region, we constructed a 1.5-Mb YAC contig mapping to 10q25 that contains MXI1 and ADRA2A as well as a number of STSs. Rare cutting restriction site analysis of overlapping YACs allowed fine mapping of these genes and markers along the contig and revealed the presence of four CpG islands. MXI1 and ADRA2A appear to be about 600 kb apart, whereas ADRB1 is separated from ADRA2A by a distance larger than previously reported. PMID- 9344666 TI - PMS2-related genes flank the rearrangement breakpoints associated with Williams syndrome and other diseases on human chromosome 7. AB - The human PMS2 mismatch repair gene and a family of at least 17 other related genes (named human PMSR or PMS2L genes) have been localized to human chromosome 7. Human PMS2 has been mapped previously to 7p22 and shown to be causative in hereditary nonpolyposis colon cancer (HNPCC), but the human PMS2L genes have not been positioned in the context of the physical or genetic map of chromosome 7. In this study we have used various mapping methodologies to determine the precise location of the human PMS2L genes at 7q11.22, 7q11.23, and 7q22. Within 7q11.23, human PMS2L genes were found to be present at at least three sites as part of duplicated genomic segments that flank the most common rearrangement breakpoints in Williams syndrome. PMID- 9344668 TI - Molecular cloning and chromosomal localization of PD-Ibeta (PDNP3), a new member of the human phosphodiesterase I genes. AB - Phosphodiesterase I (EC 3.1.4.1)/nucleotide pyrophosphatase (EC 3.6.1.9) enzymes are a family of type II transmembrane proteins that catalyze the cleavage of phosphodiester and phosphosulfate bonds of a variety of molecules, including deoxynucleotides, NAD, and nucleotide sugars. The human genes for two members of this family have been cloned and designated PC-1 (PDNP1) and PD-Ialpha/autotaxin (PDNP2). We have now cloned the third member of this family from a human prostate cDNA library and designated it human phosphodiesterase-Ibeta (PD-Ibeta). The PD Ibeta cDNA contains a 2625-bp-long open reading frame which encodes an 875-amino acid protein. COS-7 cells transfected with an expression vector, pBK-CMV, containing PD-Ibeta cDNA had high phosphodiesterase I activity compared to the mock-transfected cells. By using in situ hybridization to human metaphase chromosomes, we have assigned the locus for the PD-Ibeta (PDNP3) gene to the q22 region of human chromosome 6. PMID- 9344669 TI - Genomic imprinting in the rat: linkage of Igf2 and H19 genes and opposite parental allele-specific expression during embryogenesis. AB - Igf2 and H19 are closely linked imprinted genes in both mice and humans that are expressed from opposite parental alleles. In this study we demonstrate that these two genes are also closely linked in the rat, with the H19 gene mapping to within 145 kb of Igf2 on rat chromosome 1. We identified polymorphisms in H19 and Igf2 transcripts in two inbred rat strains and determined the expression of the parental alleles in F1 offspring. The H19 gene was shown to be expressed exclusively from the maternal allele in all fetal and neonatal tissues. Monoallelic expression of Igf2 from the paternal allele was found in all tissues except the leptomeninges and choroid plexus. Igf2 in the choroid plexus was monoallelic at days 13.5 and 15.5 of gestation with a switch to biallelic expression by day 18.5, demonstrating a loss of imprinting after the choroid plexus has differentiated. Biallelic expression of Igf2 was observed in the leptomeninges at all fetal and neonatal stages analyzed. These studies demonstrate conservation of imprinting of two closely linked genes transcribed from opposite parental alleles in a species other than human or mouse. A comparative approach between different species will be important in defining the mechanisms that regulate the tissue-specific expression of imprinted genes. PMID- 9344670 TI - Exon-intron organization of the human dystrophin gene. AB - Analysis of the exon-intron organization of the human dystrophin gene has been hampered by its enormous size. By using a YAC-based exon mapping approach and long PCR, we have succeeded in defining the size of the gene and its organization. Our results, compared with data on the distribution of deletion breakpoints by intron, elucidate the topography of the intragenic deletion-prone regions. Within the central high-frequency deletion region, the small, 6.6-kb, intron 49 shows a much higher density of deletion breakpoints than intron 44, which was previously believed to coincide with the most mutable zone of the gene. On the other hand, in the proximal part of the gene, deletion breakpoints do not preferentially occur in a few introns, but are spread over a large DNA segment containing introns 2 to 42. PMID- 9344671 TI - Sequence and chromosomal assignment of human BAPX1, a bagpipe-related gene, to 4p16.1: a candidate gene for skeletal dysplasia. AB - Homeobox-containing genes play an important role in both body axis determination and specific organ development. They have increasingly been found to be mutated in human birth defect disorders. Sequences of two bagpipe-related genes in the mouse, Nkx-3.1 and Nkx-3.2, have already been reported. Nkx-3.1 was isolated from the prostate, and its human homolog NKX-3.1 has also been described. Mouse Nkx 3.2, or bapx1, has also been isolated, and its expression in the visceral mesoderm and embryonic skeleton in the mouse has been described. We report here the human BAPX1 sequence and its localization to chromosome region 4p16.1 and suggest BAPX1 as a candidate for disorders of skeletal development that might map to 4p16.1. PMID- 9344672 TI - Genomic organization, 5'-flanking region, and chromosomal localization of the human RGS3 gene. AB - RGS3 is the largest member of a recently discovered family of proteins (RGS proteins) that appear to function as negative regulators of heterotrimeric G protein signaling. Seventeen mammalian RGS proteins have been identified by cloning or by comparison to expressed sequence tags, and several of these proteins have been shown recently to function as GTPase-activating proteins for G protein alpha subunits. Despite the intense interest in RGS proteins as physiological regulators of G-protein signaling, there is little understanding of the structure and regulation of mammalian RGS genes. Using long-distance PCR, we amplified and characterized the entire coding and 5'-untranslated region of the human RGS3 gene. The coding region of the human RGS3 gene spans 14.7 kb and contains six exons, and the 5'-untranslated region spans 3.2 kb and contains two exons. Mapping of the exons revealed that the RGS domain, conserved among all RGS proteins, was encoded by three exons, while the unique amino-terminal domain of RGS3 was encoded by a single exon. Comparison of the location of the intron-exon boundaries of the human RGS3 gene to that of the human RGS2 gene, the only mammalian RGS gene described previously, revealed a remarkable similarity, providing the first conceptual support for a common ancestral mammalian RGS gene. 5'-RACE analysis was used to map the transcription start site 517 bp upstream of the translation start site, and anchored PCR was performed to amplify 1.0 kb of genomic DNA upstream of the transcription start site. Analysis of the 5'-flanking region revealed the presence of many potential regulatory elements, the presence of an initiator (Inr) element overlapping the transcription start site, and the absence of a TATA or a CCAAT box at the usual positions. By radiation hybrid mapping, the RGS3 gene was assigned to human chromosome 9q31-q33. This study is the first to elucidate the structure, chromosomal location, and regulatory sequences of the RGS3 gene, and it establishes the genetic basis for RGS3 gene research in humans. PMID- 9344673 TI - Molecular cloning and characterization of the human mitochondrial NADH:oxidoreductase 10-kDa gene (NDUFV3). AB - The human gene for the 10-kDa flavoprotein subunit of the mitochondrial NADH:ubiquinone oxidoreductase (Complex I) was completely cloned and sequenced. The so-called NDUFV3 gene contains three exons, spanning 20 kb. The open reading frame contains a 34-codon import sequence and a 74-codon mature protein sequence. A database search revealed close homology to bovine and rat protein sequence but not to any other known protein. Northern blot analysis showed that the NDUFV3 gene is ubiquitously expressed. The NDUFV3 gene was assigned by FISH to a single location on chromosome 21q22.3 and might contribute to the Down syndrome phenotype. PMID- 9344674 TI - Genomic sequence and organization of the human gene for cytochrome c oxidase subunit (COX7A1) VIIa-M. AB - Cytochrome c oxidase (COX, EC 1.9.3.1), the last component of the mitochondrial electron transfer chain, is built up by 13 polypeptides; 3 of them are encoded by the mitochondrial genome while the 10 smaller subunits are encoded by the nuclear genome. Several nuclear-encoded subunits occur in two different tissue-specific isoforms, a constitutive "L"-form and an "M"-form specific for skeletal and heart muscle. In this article, we describe the genomic sequence and organization of the human gene for COX subunit VIIa-M (COX7A1) located on chromosome 19q13.1 and compare it to its bovine homologue. The coding region of the gene extends over 1.45 kb of genomic sequence, organized in four exons. Intron-exon boundaries are well conserved between cattle and humans. Although it is a gene for a tissue specific isoform, it has some features of a housekeeping gene: it is located in a CpG island, like its bovine homologue, and no TATA or CCAAT boxes were found in the 5' flanking sequence. Southern hybridization of COX7A1 to human genomic DNA revealed no pseudogenes. Putative binding sites for ubiquitous transcription factors like Sp1 and specific expression in skeletal as well as in heart muscle have been found. In contrast to the bovine gene, the human gene contains putative binding sites for nuclear respiratory factor 2 (NRF-2), which is implicated in the activation of other respiratory enzymes. Therefore, the human and the bovine genes, although well conserved in their coding regions, could differ in the tissue-specific regulation of gene expression. Knowledge of the gene structure will facilitate the analysis of the involvement of subunit VIIa in mitochondrial myopathies and may provide clues to the function of this subunit in a multicomponent enzyme. PMID- 9344675 TI - Cloning, structural organization analysis, and chromosomal assignment of the human gene for the neurosecretory protein VGF. AB - The Vgf gene was originally identified as a 2.7-kb cDNA fragment isolated from nerve growth factor-treated PC12 cells by differential display against PC12 cells. It is transcribed solely in subpopulations of neuroendocrine cells in vivo and it is induced by neurotrophins in target cells in vitro. The single-copy human VGF gene was isolated from a genomic library. The gene spans approximately 6 kb and contains two exons. The entire VGF protein is encoded by exon 2, while exon 1 contains only 5'-untranslated sequence. The structural organization of the human gene is similar to that described for the rat Vgf gene (S. R. J. Salton et al., 1991, Mol. Cell. Biol. 11: 2335-2349) and both the translated and the untranslated regions show a high degree of sequence homology to the rat gene. Northern blot analysis revealed a single transcript of approximately 2.7 kb that was detected only in mRNA preparations from brain. The gene was assigned to chromosome 7q22 by fluorescence in situ hybridization. PMID- 9344676 TI - The rapidly evolving Pem homeobox gene and Agtr2, Ant2, and Lamp2 are closely linked in the proximal region of the mouse X chromosome. AB - The Pem gene encodes a homeodomain-containing protein expressed in reproductive tissue that may function as a transcription factor regulating spermatogenesis and sperm maturation. We have mapped the Pem gene to the proximal end of the mouse X chromosome, placing it within the Hprt region. Based on the mapping of Pem and other loci in three separate Mus musculus x Mus spretus backcross panels, we established the order of markers within this segment of the Hprt region as: Agtr2 Pem-Ant2-DXMit50-Lamp2-DXMit49. In contrast to some other regions of the X chromosome, which have been rearranged during the evolution of mammals, we show that the order of gene loci within this Hprt region is conserved in mice and human. The finding that the mouse Ant2 and Pem loci are tightly linked suggests that human ANT2 may be useful as a marker for isolating the human PEM gene, which has been impervious to cloning by conventional hybridization methods because of its rapid evolution. PMID- 9344677 TI - Characterization of a KRAB family zinc finger gene, ZNF195, mapping to chromosome band 11p15.5. AB - We report the cDNA sequence of the zinc finger gene, ZNF195, which maps to chromosome 11p15.5. ZNF195 contains an N-terminal KRAB domain and 14 tandemly repeated Kruppel type zinc finger motifs at its C-terminus. Northern analysis shows expression of ZNF195 in adult heart, brain, placenta, skeletal muscle, and pancreas with a predominant transcript size of 4.3 kb. There is little expression in adult lung, liver, and kidney. In fetal lung, liver, kidney, and brain, the predominant transcript is 3.5 kb. Fetal brain also expresses a 4.3-kb transcript. RT-PCR analysis shows that two exons, 4a, which contains an inverted Alu sequence, and 4b, are differentially spliced and absent from the major transcript. PMID- 9344678 TI - The Novel Epithelial-Specific Ets Transcription Factor Gene ESX Maps to Human Chromosome 1q32.1 PMID- 9344680 TI - Assignment of the human equilibrative nucleoside transporter (hENT1) to 6p21.1 p21.2. PMID- 9344679 TI - Radiation hybrid mapping of human ADAM10 gene to chromosome 15. PMID- 9344681 TI - Localization of the gene for a serine protease with IGF-binding domain (PRSS11) to human chromosome 10q25.3-q26.2. PMID- 9344682 TI - Assignment of the mouse Hsp25 and Hsp105 genes to the distal region of chromosome 5 by linkage analysis. PMID- 9344683 TI - MEETING REPORT PMID- 9344684 TI - Guidelines for human gene nomenclature (1997). HUGO Nomenclature Committee. PMID- 9344685 TI - Rules and guidelines for mouse gene nomenclature: a condensed version. International Committee on Standardized Genetic Nomenclature for Mice. PMID- 9344686 TI - Androgen receptor (AR) immunoreactivity in rat pudendal motoneurons: implications for accessory proteins. AB - Pudendal motoneurons in male rats are located in two sexually dimorphic motoneuronal pools: the spinal nucleus of the bulbocavernosus (SNB) and the dorsolateral nucleus (DLN). SNB motoneurons innervate sexually dimorphic muscles bulbocavernosus (BC) and levator ani (LA) and the sexually monomorphic external anal sphincter (EAS) muscle. DLN motoneurons innervate either the sexually dimorphic ischiocavernosus (IC) muscle or the sexually monomorphic external urethral sphincter (EUS) muscle. Previous observations indicate that the size of BC, LA, and IC motoneurons in males is sensitive to adult androgen manipulations, whereas the size of EAS and EUS motoneurons is not, raising the question of whether this difference in androgen sensitivity among pudendal motoneurons reflects a difference in androgen receptor (AR) expression. AR immunocytochemistry using the PG-21 antiserum was carried out on spinal cord tissue from normal adult male rats in which specific pudendal motoneuronal subpopulations were identified with retrograde markers. Over 90% of BC, LA, and IC motoneurons displayed AR immunoreactivity in their nuclei. Among motoneurons in the SNB, significantly fewer EAS motoneurons had AR-positive nuclei, which may contribute to the reported failure of EAS motoneurons to morphologically respond to changes in androgen levels. However, within the DLN, despite the fact that IC but not EUS motoneurons are reported to respond to androgen with an increase in soma size, IC and EUS motoneurons had the same proportion of AR-positive nuclei. These results indicate that androgen receptors, while necessary, are not sufficient to confer androgen sensitivity to cells. PMID- 9344687 TI - Central administration of chicken gonadotropin-releasing hormone-II enhances courtship behavior in a female sparrow. AB - Like most vertebrates, birds have two forms of gonadotropin-releasing hormone (GnRH). Chicken GnRH-I (cGnRH-I) is released at the median eminence to elicit gonadotropin release; chicken GnRH-II (cGnRH-II) is thought to be non hypophysiotropic and its function is unclear. Both forms are hypothesized to act as neurotransmitters in the control of reproductive behavior. In the present study, we implanted chronic cannulae aimed at the third ventricle in female white crowned sparrows (Zonotrichia leucophrys gambelii) to test the effects of both forms of GnRH on copulation solicitation, a female courtship behavior. This behavior can be elicited in captive, estrogen-primed females by playing a recording of male song. We quantified the behavioral response to recorded song 30, 60, and 90 min after intracerebroventricular infusion of cGnRH-I, -II, or saline. cGnRH-II, but not cGnRH-I, increased solicitation behavior compared with saline 30 min after infusion. Under control conditions, responses to the playback diminish from the 30-min to the 90-min time point. Responses after cGnRH-II infusion followed a similar pattern, whereas after cGnRH-I, there was no significant response decrement. cGnRH-I appears to maintain the level of solicitation seen at 30 min after infusion. Our results suggest a behavioral role for cGnRH-II that may be independent of cGnRH-I. PMID- 9344688 TI - Deficits in reproductive behavior in diabetic female rats are due to hypoinsulinemia rather than hyperglycemia. AB - These studies determined whether deficits in reproductive behavior observed in streptozotocin (STZ)-induced diabetic female rats are caused by hyperglycemia or loss of insulin. Female Sprague-Dawley rats were ovariectomized and made diabetic by a single ip injection of STZ (75 mg/kg). Reproductive behavior was measured 12 days after the onset of hyperglycemia following the injection of estrogen and progesterone in doses known to restore reproductive behavior in nondiabetic rats. Rats in which STZ produced diabetes showed significantly reduced receptive and proceptive sexual behaviors. Normalization of blood glucose levels either by restricting diet or by phlorizin treatment failed to restore reproductive behavior in diabetic animals. However, even doses of insulin which were not fully effective in correcting peripheral hyperglycemia were able to prevent the STZ induced behavioral deficit. No changes in general activity were observed in any experimental group as assessed by open field activity. The density of the norepinephrine transporter, as measured by [3H]nisoxetine binding, was reduced in the cortex but not in the brain stem, hypothalamus, or hippocampus of diabetic animals. Insulin treatment prevented the loss of cortical [3H]nisoxetine binding, and even partial normalization of blood glucose restored cortical [3H]nisoxetine binding to control levels. These findings suggest that diabetes-induced reproductive deficits are due to hypoinsulinemia and cannot be corrected simply by the normalization of blood glucose, whereas reductions in the density of cortical norepinephrine transporter result from hyperglycemia. PMID- 9344689 TI - Social environment and steroid hormones affect species and sex differences in immune function among voles. AB - Testosterone has bipotential effects on male fitness; that is, it both suppresses immune function and maintains characteristics important for reproductive success. Presumably, these effects of testosterone may be more pronounced among polygynous species because testosterone concentrations are generally higher among polygynous than monogamous males. The present study examined sex and species differences in cell-mediated immunity among four arvicoline rodents. The role of mating system and sex steroids in sex differences in immune function was examined in individually housed polygynous meadow (Microtus pennsylvanicus) and montane (M. montanus) voles and monogamous prairie (M. ochrogaster) and pine (M. pinetorum) voles in Experiment 1. No sex differences in splenocyte proliferation were observed among the four species and circulating testosterone concentrations did not correlate with immune function of individuals within each species. The contribution of social isolation to these results was examined in Experiment 2, in which meadow and prairie voles were housed individually, or with same- or opposite-sex conspecifics in either pairs or groups of four per cage for 28 days. Overall, prairie voles exhibited more robust immune responses than meadow voles when housed in pairs or in same-sex groups. Sex differences in immune function were also apparent; male meadow voles had higher immune responses than female conspecifics when housed in pairs, whereas female prairie voles had higher responses than male conspecifics when housed in same-sex pairs. Circulating sex steroid hormones and corticosterone appear to mediate some, but not all, of the changes in immune function evoked by differential housing conditions. Taken together, these results suggest that social factors have significant effects on immunity and should be considered in studies of sex differences in immunity at both proximate and ultimate levels. PMID- 9344690 TI - Thinking about networks in the control of male hamster sexual behavior. AB - Motivated social behaviors such as mating are controlled by a complex network of limbic nuclei. Concepts of network organization derived from computational neuroscience may aid our understanding of the links between the neuroanatomical circuitry and what is represented by the anatomy. Research in my laboratory uses mating behavior in the male Syrian hamster as a model to elucidate how chemosensory and steroid cues are integrated in the brain. An interaction of odors and hormones is required for mating in this species. These two essential stimuli are transmitted through separate parallel pathways in the limbic system. The functional organization of the hamster mating behavior circuit is characterized by distributed representation, divergent and convergent neural pathways, and recurrent feedback. Odors and hormones have different modes of action on this neural network. While chemosensory cues stimulate the input units of the network, steroids facilitate behavior through the hidden units. In this manner, steroids appear to create a permissive environment for subsequent activation by odor cues. PMID- 9344691 TI - The effects of testicular tissue and prehatching inhibition of estrogen synthesis on the development of courtship and copulatory behavior in zebra finches. AB - As in many mammalian and avian species, testicular androgens or their metabolites activate courtship and copulatory behaviors in adult male zebra finches. However, studies of sexual differentiation of these behaviors and related anatomical structures provide conflicting results. For example, posthatching estradiol can both masculinize courtship and the neural structures involved in song in females and inhibit the development of masculine copulation in males. These and other results have led to the hypotheses that (1) testicular androgens are converted to estradiol in the brain of developing males, and estradiol serves to masculinize the song system, whereas (2) estradiol secretion by the female ovary allows feminine rather than masculine copulatory behavior to develop. Treating embryonic zebra finches with the estrogen synthesis inhibitor fadrozole causes functional testicular tissue to develop in genetic females. The present study investigated the effects of such treatment on the development of singing and copulatory behavior as well as song system anatomy in males and females. While exogenous testosterone facilitated the display of sexual behaviors in adult males, the testicular tissue in females had no masculinizing effect on the production of audible courtship song or copulation. Their song control nuclei were also not masculinized, even in individuals lacking ovarian tissue. In contrast, embryonic inhibition of estrogen synthesis in males significantly stimulated song production. These results suggest that while manipulations of steroid hormone exposure can influence the display of sexual behaviors, gonadal secretions may not be required for normal sexual differentiation of the song system in zebra finches. PMID- 9344692 TI - Lordosis in male rats: effect of dorsal raphe nucleus cuts. AB - The efferents and/or afferents of the dorsal raphe nucleus (DRN) were transected by several types of cut in castrated male rats, and lordosis behavior was observed after implantation with Silastic tubes containing estradiol. Throughout the behavioral tests, low incidences of lordosis were observed in control male rats without brain surgery or with a sham operation. In contrast, all male rats with a horizontal circle cut at the ventral area of the DRN displayed lordosis, and the mean lordosis quotient (LQ) was higher than that in control rats, while rats with a horizontal cut at the dorsal area of the DRN did not. Furthermore, mean LQs in male rats with an anterior half-circle horizontal cut at the ventral area of the DRN were higher than those in control groups. A posterior half-circle cut at the ventral area had no effect. In addition, male rats with a half-dome cut located anterior to the DRN showed a high LQ score, but rats with a posterior half-dome cut did not. These results suggest that anterior and anteroventral neural fibers of the DRN are involved in the lordosis inhibiting mechanism in male rats. PMID- 9344693 TI - Mucosal tolerance: a two-edged sword to prevent and treat autoimmune diseases. AB - Mucosal administration of autoantigens results in the development of a state of peripheral immunological tolerance. Depending upon the dose of antigen administered, anergy/deletion of antigen-specific T cells (higher doses) and/or selective expansion of cells producing immunosuppressive cytokines (TGF-beta, IL 4, and IL-10) (lower doses) are two major mechanisms in mucosal tolerance induction. Mucosal tolerance is more effective after nasal compared to oral administration of antigens at the same dose. A large series of studies have demonstrated that mucosal tolerance by oral or nasal antigen administration effectively prevents several experimental disease models (EAE, EAMG, EAN, EAU, IDDM, and CIA). Mucosal antigen administration is superior in prevention to treatment of autoimmune diseases. To broaden the effectiveness of mucosal tolerance, a conjunction of tolerogens with cytokines/CTB might enhance suppression of clinical disease. Based on experimental experience with mucosal tolerance, trials in humans are ongoing in MS, RA, and uveitis. However, mucosal tolerance induction is related to the route of antigen administration (oral, nasal, parentetal), type of antigen (whole protein, peptide, altered peptide), and timing with regard to disease onset and may represent a two-edged sword. In particular, the risks of worsening an ongoing autoimmune disease by mucosal antigen administration have been incompletely addressed. Here we give an overview on some recent developments in this field where, however, much more studies are needed to define an ultimate and safe procedure. PMID- 9344694 TI - Substance P: a regulatory neuropeptide for hematopoiesis and immune functions. PMID- 9344695 TI - T-Cell epitope mapping the PorB protein of serogroup B Neisseria meningitidis in B10 congenic strains of mice. AB - T-cell epitope mapping the meningococcal serotype 15 PorB protein performed in this study in three congenic strains of mice with B10 genetic background revealed at least three murine T-cell epitopes (55-72, 163-180, and 226-261), located in the highly conserved putative transmembrane regions of Neisserial porins. Proliferation assays with popliteal lymph node cells derived from mice immunized with the PorB protein or with synthetic 18-mer peptides showed that epitope 163 180 immunized only in the H-2d haplotype, epitope 55-72 could be presented by both H-2f and H-2s molecules, while the 226-261 region covered by three overlapping peptides could be efficiently recognized in context of all three MHC class II haplotypes studied. Inhibition experiments with blocking I-Aalpha- and I Ealpha-specific mAb showed that peptide 163-180 was presented by I-Ad and peptide 244-261 was presented by both I-Af and I-As. In addition, evidence was obtained that peptide 226-243 was presented in context of H-2d or I-As haplotypes and peptide 55-72 was presented in context of I-Af and I-As loci. Finally, the Norwegian outer membrane vesicle vaccine, but not the purified PorB protein, could recall responses in mice immunized with synthetic peptides corresponding to the 226-261 region. Altogether, these results suggest that T-cell epitopes identified on the serotype 15 PorB protein, particularly those presented by several MHC class II molecules (e.g., 226-261), could have important implications for the development of meningococcal vaccines. PMID- 9344696 TI - CD5+ and CD5- B1-like lymphocytes in healthy guinea pig. AB - Spleen, lymph node, and peripheral blood lymphocytes from healthy guinea pigs (gp) were examined for their ability to produce polyreactive autoantibodies to a battery of self-antigens and to cryptic determinants (phosphatidylcholine) on bromelain-treated mouse red blood cells (Br-MRBC). The mouse monoclonal antibody (Mab) 8BE6 anti-gp pan-T (CD5) marker was used for identification of CD5+ B1 cells by the plaque-forming assay (PFC), immunofluorescence, complement-mediated cytotoxicity, and immunocytochemistry. The detection of CD5+ cells by the 8BE6 Mab depended on the method used. They were better demonstrated by cytolysis and immunocytochemistry than by FACS analysis. By the latter method, the level of the CD5+ B cell subpopulation was associated neither with the age of the gp nor with the organ examined. Similarly wide ranges of PFC were detected in untreated or LPS-treated animals regardless of age and organ. The vast majority of the LPS stimulated IgM antibody-secreting B lymphocytes reacting with the Br-MRBC, and those producing natural autoantibodies, did not bind the 8BE6 Mab. PMID- 9344697 TI - Selective expansion of gammadelta T cells among liver-derived lymphocytes of AIDS patients with disseminated Mycobacterium avium complex infection. AB - The aim of this study was to investigate a potential expansion of gammadelta T cells in AIDS patients in response to disseminated infection with Mycobacterium avium complex. Liver-derived lymphocytes and peripheral blood lymphocytes were obtained from 5 AIDS patients with disseminated Mycobacterium avium complex infection and 10 HIV-infected patients without mycobacterial disease. Control patients included 14 HIV-negative patients without bacterial disease and 4 HIV negative patients with disseminated Mycobacterium tuberculosis infection. The percentage of gammadelta T cells among CD3+ T lymphocytes was determined by flow cytometry. gammadelta T cells were markedly enhanced in liver biopsies but not among peripheral blood lymphocytes of AIDS patients with disseminated M. avium complex infection (median liver gammadelta/CD3: 26%) as compared to HIV-infected control patients without mycobacterial disease (median liver gammadelta/CD3: 2.3%; P < 0,005). Disseminated infection with M. tuberculosis in HIV-negative patients leads to a similar expansion of gammadelta T cells among liver-derived CD3+ lymphocytes (median gammadelta/CD3: 15.5%) as compared to control patients without mycobacterial disease (median gammadelta/CD3: 2.15%; P < 0,001). PMID- 9344698 TI - Inflammatory cells and MHC class II antigens expression in prostate during time course experimental autoimmune prostatitis development. AB - The degree of lymphocytic infiltration alongside the phenotype of the infiltrating cells and MHC class II expression were studied in rats during a time course experimental autoimmune prostatitis (EAP) development. Inflammatory foci per square millimeter were scarce at day 7 after first immunization (FI) and were composed of few mononuclear cells. The number of inflammatory foci per square millimeter increased at day 14 and remained with slight variations at days 21 and 28 after FI. The number of mononuclear cells per square millimeter increased on day 14, diminished slightly on day 21 and reached the highest level on day 28. All these infiltrates were constituted by CD4 and CD8 T cells whereas only few macrophages were present. Mast cells were also present reaching maximum levels on day 7 after FI and then decreased. MHC class II antigens were found in epithelial cells during EAP development. IA showed a similar pattern in all periods analyzed whereas IE showed a modulating behavior, reaching the highest expression on day 21 after FI. In this experimental model, the differential expression of MHC class II antigens could modulate the immune response during EAP development. PMID- 9344699 TI - Complement activation in severe Plasmodium falciparum malaria. AB - We determined indices of plasma complement activation (C3, C4, Bb, C4d, iC3b, and SC5b-9), levels of tumor necrosis factor (TNF) and interleukin-6, and the APACHE II score in 23 patients with complicated Plasmodium falciparum malaria. On admission, plasma concentrations of Bb, SC5b-9, and C4d were markedly increased compared to healthy control subjects (n = 24) (4.5 +/- 1.9 vs 1.5 +/- 0.6 mg/L; 1125.7 +/- 496.9 vs 183.2 +/- 76.5 microg/L; and 15.7 +/- 5.7 vs 7.2 +/- 1.4 mg/L, P < 0.01 for all). In contrast C3 and iC3b concentrations were decreased (631.4 +/- 247 vs 947.3 +/- 243.2 and 105 +/- 17.9 vs 151.3 +/- 14.5 mg/L; P < 0.01 for both). Plasma C4 concentrations in malaria were not different from normal controls. Plasma Bb, C3, and iC3b levels normalized on day 7 of treatment, whereas SC5b-9 and C4d levels remained elevated. A significant correlation between elevated TNF levels and Bb (r = 0.507) and SC5b-9 (r = 0.448, P < 0.01 for both) and a negative correlation between iC3b and SC5b-9 and TNF levels existed (r = -0.537 and r = -0.466, P < 0.01 for both). In addition, a significant correlation between C3 and iC3b (r = 0.689) and C4 and C4d (r = 0.737) existed. However, no relation between clinical disease severity and complement fragments existed. The results demonstrate that both the classical and the alternative pathways of the complement system are profoundly activated in complicated malaria. PMID- 9344700 TI - Development of a rapid whole blood flow cytometry procedure for the diagnosis of X-linked hyper-IgM syndrome patients and carriers. AB - The CD40 ligand expressed on activated T cells plays a pivotal role in B cell proliferation and differentiation. Mutations in the CD40 ligand gene, which alter its expression on the surface of activated T cells, are associated with the X linked form of Hyper-IgM syndrome (XHIM). A rapid and simple, three-color whole blood flow cytometry procedure was developed for maximal expression and detection of the CD40L on the surface of in vitro activated CD4+ T cells. Approximately 90% of in vitro activated CD4+ T cells obtained from healthy controls expressed the CD40L compared to only 5% of in vitro activated CD4+ T cells obtained from the XHIM patients. The CD40L was expressed on approximately 50% of the in vitro activated CD4+ T cells obtained from the mothers of XHIM patients, consistent with a diagnosis of their carrier status. This is the first report of a whole blood procedure adapted for routine clinical use which is able to detect abnormal CD40L expression in XHIM patients and carriers. PMID- 9344701 TI - Diagnostic significance of antibodies to glutamic acid decarboxylase in Japanese diabetic patients with secondary oral hypoglycemic agents failure. AB - Some non-insulin-dependent diabetes mellitus (NIDDM) patients are positive for antibodies to glutamic acid decarboxylase (anti-GAD), and they tend to develop insulin deficiency. The aim of this study was to evaluate the prevalence of anti GAD in NIDDM with secondary failure of sulfonylurea agents (NIDDM-SF) and to investigate the diagnostic significance of seropositivity for anti-GAD in NIDDM SF patients by evaluating human leukocyte antigen (HLA)-DRB1 alleles concurrently. The prevalence of anti-GAD in NIDDM-SF, NIDDM, and new-onset (within 1 year after onset) insulin-dependent diabetes mellitus (IDDM) was 9.3% (39/420), 3.1% (12/392), and 65.0% (13/20), respectively. Pancreatic beta cell function deteriorated in NIDDM-SF patients positive for anti-GAD. HLA-DRB1 allele typing revealed that NIDDM-SF patients positive for anti-GAD were significantly associated with DRB1*0901 (RR = 2.81, P < 0.01), which is one of the susceptible alleles to IDDM. Shorter interval before development of secondary failure and insulin deficiency were significantly associated with the presence of DRB1*0901 (P < 0.05) in NIDDM-SF patients positive for anti-GAD. In conclusion, nearly 10% of NIDDM-SF patients are positive for anti-GAD, suggesting that an autoimmune mechanism might play an important role in the pathogenesis of NIDDM-SF patients. In addition, a combination of serological marker (anti-GAD) and genetic marker (HLA-DRB1) is useful for predicting clinical course of NIDDM patients with secondary failure of sulfonylurea agents. PMID- 9344702 TI - Primary hormonogenic sites as conserved autoepitopes on thyroglobulin in murine autoimmune thyroiditis: role of MHC class II. AB - A few synthetic peptides corresponding to amino acid sequences on human thyroglobulin (Tg) have been reported to induce moderate thyroiditis or activate mouse Tg (MTg)-primed T cells to transfer thyroiditis in mice susceptible to experimental autoimmune thyroiditis. Using three pairs of 12-mer peptides (1-12, 2549-2560, 2559-2570), with thyroxine (T4) or noniodinated thyronine (T0) at the conserved, hormonogenic site 5, 2553, or 2567 respectively, we reported that iodination was not required for a Tg hormonogenic site to be a thyroiditogenic autoepitope. To determine the relative importance of MHC class II and T cell receptor (TCR) repertoire, we compared two EAT-susceptible k and s (CBA and A.SW) haplotypes and their respective MHC-identical strain (C57BR and SJL) with approximately 50% genomic deletion of TCR Vbeta genes. Whereas k and s strains develop MTg-induced EAT, vigorous immunization with peptides containing T4 or T0 at either 5 or 2553, but not at 2567, led to mild (10-20%) thyroiditis only in some mice of either k strain. TCR Vbeta gene differences played a minor role. T cell responses to all peptide pairs were quite similar in CBA and C57BR mice, and both hT0(2553) and hT4(2553) reciprocally primed and stimulated their T cells. In adoptive transfer, SJL mice were somewhat more responsive to peptide activation than A.SW but much weaker than k strains. By comparing T4- and T0-containing peptides in different haplotypes, we show further that antigenicity of conserved hormonogenic sites is intrinsic, dependent more on amino acid sequence and binding to appropriate class II molecules and less on TCR repertoire or iodination of T0. PMID- 9344703 TI - Signals transduced through the CD4 molecule interfere with TCR/CD3-mediated ras activation leading to T cell anergy/apoptosis. AB - It has been previously demonstrated that the occupancy of CD4 molecules by the HIV-1 envelope glycoprotein gp120 results in marked inhibition of T cell receptor CD3 complex (TCR/CD3) activation-induced IL-2 secretion. To elucidate the mechanism of inhibitory effects of gp160 on T cell signaling, we have investigated the intracellular biochemical events and biological output in response to anti-CD3 mAb activation of purified peripheral blood CD4+ T cells from healthy donors with and without prior exposure to HIV-1 gp160. Pretreatment with gp160 resulted in marked inhibition of tyrosine phosphorylation of p59(fyn), PLC-gamma1, ras activation, and TNF-alpha secretion in anti-CD3 mAb activated CD4+ T cells, and a subset of CD4+ cells underwent activation-induced cell death. The data presented here provide insight into the mechanism by which the interaction of HIV-1 envelope glycoproteins with CD4 molecules may alter TCR/CD3 activation-induced signal transduction resulting in anergy and apoptosis with consequent functional deficiency of CD4+ T cells. PMID- 9344704 TI - Inhibition of the progression of multiple sclerosis by linomide is associated with upregulation of CD4+/CD45RA+ cells and downregulation of CD4+/CD45RO+ cells. AB - In a recent double-blind, phase II study, conducted in our department, we showed that Linomide-treated MS patients had significantly less active lesions (in serial monthly MRI tests) and a tendency for clinical stabilization. Here we present the immunological evaluation of the patients who participated in this study and propose a novel mechanism by which Linomide downregulates autoreactivity. Peripheral blood leukocytes (PBLs), serum, and CSF samples were obtained at two to four time points over the 6 months of the trial. Flow cytometric analysis (FACS) of the CD5/CD19, CD4/CD8, CD14/CD3, CD16/CD3, CD45RA/CD4, and CD45RO/CD4 surface markers on PBLs was performed and the levels of the IL-1beta, IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma, and IL-2R were also examined. White blood counts of Linomide-treated patients were consistently elevated throughout the treatment period (P = 0.002-0.04). Cytokines levels in serum and CSF were highly fluctuating and we could not detect any clear trend as a result of Linomide treatment. FACS analysis showed that Linomide treatment significantly increased the percentage of the CD4+/CD45RA+ cells (from 35.5% at baseline to 42.3% at week 24; P = 0.02), and decreased CD4+/CD45RO+ lymphocytes (62.6% at baseline vs 53.7% at week 24, P = 0.02). Linomide also induced a transient increase in the NK-cells, the NK 1.1 cells, and the CD5 B-cells (P = 0.02). Upregulation of naive CD45RA T-lymphocytes and parallel downregulation of memory CD45RO cells seems to be one of the main mechanisms by which Linomide inhibits MS activity and may represent an alternative immunomodulating approach for the treatment of MS and autoimmunity in general. PMID- 9344706 TI - Cloning and characterization of murine thyroglobulin cDNA. AB - Thyroglobulin is used to induce in mice experimental autoimmune thyroiditis (EAT), a model for Hashimoto thyroiditis. Because murine thyroglobulin is a more potent inducer of EAT than heterologous thyroglobulins, it has been hypothesized that it contains unique pathogenic epitopes. The validation of this hypothesis has been hampered by the lack of the murine thyroglobulin sequence. To identify murine-specific areas in thyroglobulin, we cloned, by reverse transcriptase PCR, and sequenced the complete murine thyroglobulin cDNA. This encodes a polypeptide of 2748 amino acids that is 73.5 and 71.8% identical to bovine and human thyroglobulin, respectively. Six regions are unique to each species. We also analyzed through EpiMer the sequences able to bind to the I-Ek major histocompatibility allele and, therefore, function as T cell epitopes. EpiMer analysis showed seven murine-specific T cell epitopes in thyroglobulin. The availability of the complete murine thyroglobulin sequence should promote the understanding of the pathogenesis and immunoregulation of EAT. PMID- 9344705 TI - Low-level production of interleukin-13 in synovial fluid and tissue from patients with arthritis. AB - Rheumatoid arthritis (RA) is a chronic, aggressive disease characterized by inflammatory cells in the synovial tissue (ST) and synovial fluid (SF). Interleukin (IL)-13 inhibits the production of proinflammatory cytokines, chemokines, and hematopoietic growth factors by activated human monocytes. The aim of this study was to determine the production of IL-13 in various forms of arthritis. The presence of IL-13 in RA was found to be low, in that 18 of 26 RA SF samples and 10 of 14 RA peripheral blood (PB) samples had nondetectable levels ( LEW and LEW > ACI small bowel transplant models. Animals were divided into treatment groups: I, none; II, Allotrap (20 mg/kg/day on Days 0 4); III, cyclosporine (CsA; 10 mg/kg/day on Days 0-4); IV, Allotrap + CsA (as in groups II and III); V, Allotrap (40 mg/kg/day every other day on Days -19 to 4); VI, Allotrap + CsA (as in groups III and V); VII, Allotrap + CsA (as in groups III and V, with Allotrap administered intragraft Days 0-4). The animals were sacrificed at the time of graft rejection (defined by dusky, necrotic stoma and increased stomal output). Peripheral blood, spleen, native bowel, and allograft intraepithelial and lamina propria lymphocytes were harvested and mixed lymphocyte culture (MLC) reactivity against self, donor, and third-party splenocytes was assessed. Statistical analysis was performed by ANOVA with Dunnett's t for multiple comparisons against a control as a post hoc test. We found a very slight, but significant prolongation of graft survival in with treatment protocol V for both strain combinations. In addition, MLC response of splenocytes to donor antigen was decreased with combined CsA and Allotrap, but not with Allotrap alone. We conclude that Allotrap decreases response to alloantigens, and slightly, but significantly prolongs graft survival in the hihgly immunogenic small bowel transplant model. PMID- 9344718 TI - Phase-directed therapy and cardiac xenograft survival. AB - Xenotransplant rejection is facilitated not only by T cell upregulation but also by endothelial activation and B cell/antibody mechanisms, which standard immunosuppression is unable to overcome and xenorejection ensues. However, therapy directed specifically at each phase of xenorejection may improve xenograft survival. To study this we used a heterotopic cardiac xenotransplant mode (Syrian hamster to Lewis rat). Controls had no immunotherapy. Xenorecipients received cyclosporine to restrict T cellular response/development or cyclophosphamide, an antiproliferative, to reduce xenoreactive clones and antibody/complement injury, or anti-TNF antibody to alter cytokine cascades and endothelial activation/inflammation. Further xenorecipients received combinations. While single modalities alone did not enhance survival, combinations appeared to be at least additive in vivo, suggesting that therapy directed at specific phases of xenorejection may prove useful. PMID- 9344719 TI - Towards a General Theory of Biological Signaling AB - Current models of biological communication point at evolutionary mechanisms of particular signal types. Those that present complete models look at the signals' equilibrium values and their evolutionary stability, and require two simultaneous equations: an equation that describes the signaler's fitness as a function of the signal and of the recipients' response, and a simultaneous equation that represents the fitness of recipients. This paper examines the effect of different signal types, such as handicaps, amplifiers, camouflage, mimicry etc, on the first equation. By considering parameters that affect the evolution of signals this paper first constructs a general model of biological signaling. Different signal types are then characterized by different sets of limiting assumptions. As a result, the fitness of a signaler of each signal type is represented by a unique equation that is a mathematical derivation of the general signaling model. This analysis enables a natural division of signals into groups and subgroups that share similar assumptions and properties. It shows the importance of signal design, and points at three methods by which signals may be reliable: by trade offs between cost and benefits, by design and by convention. Copyright 1997 Academic Press Limited PMID- 9344720 TI - Allelopathy in Spatially Distributed Populations AB - In a homogeneously mixing population of E. coli, colicin-producing and colicin sensitive strategies both may be evolutionarily stable for certain parameter ranges, with the outcome of competition determined by initial conditions. In contrast, in a spatially-structured population, there is a unique ESS for any given set of parameters; the outcome is determined by how effective allelopathy is in relation to its costs. Furthermore, in a spatially-structured environment, a dynamic equilibrium may be sustained among a colicin-sensitive type, a high colicin-producing type, and a "cheater" that expends less on colicin production but is resistant. Copyright 1997 Academic Press Limited PMID- 9344721 TI - Electric Potential and Concentration of Ion Species in the Proximity of a Cell Membrane: ab initio Calculations AB - The mathematical model of ion transport in the field of electric forces and diffusion gradients is based on particle conservation equations and the Poisson equation which governs the distribution of the electric potential. Designed for evaluating the currents through passive ion channels in cellular membranes, the model is formulated in a two-dimensional approximation assuming rotational symmetry. Three ionic species are considered in the present study: sodium, calcium and chlorine. The program developed for the numerical solution is based on a semi-analytical approximation suggested by Gummel & Scharfetter. The present contribution reports calculations of the steady-state distributions of ionic species induced by an electric field due to a negative potential drop across the cellular membrane, which represent initial conditions for future calculations of the time development of the ion flux through the calcium channel. The numerical calculations were performed from the moment of potential application up to the time 6.2 MUs, when a new steady state is established. At this stage, the maximum values of the sodium and calcium concentrations at the membrane surface are about 22% and 47% above the initial values of 145 mM and 1 mM, respectively, while the chlorine concentration is approximately 18% below. The electric field at the membrane surface is about 6.7x10(4) and decreases exponentially with distance. The length of the boundary layer is about 40 A. Since the model is based on fundamental principles, it can be used for the quantitative solution of any problem possessing rotational symmetry where a continuum approach is applicable and some elementary conditions are fulfilled, such as equilibrium (or at least quasi-equilibrium) of the particle energy distributions with respect to time and electric field, and the assumption that the transport coefficients are defined as functions of the field and their numerical values are known. Copyright 1997 Academic Press Limited PMID- 9344722 TI - Assessment of g-dependent Cellular Gravitaxis: Determination of Cell Orientation from Locomotion Track AB - Movement of cells in the gravity field is principally affected in two ways: velocity and orientation. Experimental observation of gravitaxis in large cell populations can document the velocity and orientation of swimming tracks, but orientations of individual cells are not represented at low magnifications. Cell orientations may depart from track orientations due to superposition of sedimentation on cellular propulsion. Here, we show that determination of the sedimentation rate in addition to cell track parameters allows a reconstitution of cell orientation employing geometric principles. Published and original cellular data indicate that gravitactic orientation of cells swimming in the gravity field is superior to that suggested from the experimental tracks. Similar conclusions apply to cells which walk or glide along substrate surfaces. Calculation of cell orientation coefficients provides a basis for determinations of the acceleration-dependence of gravitaxis and for quantitative tests on physical and/or physiological principles of cellular gravitaxis. Copyright 1997 Academic Press Limited PMID- 9344723 TI - Evolutionarily Stable Reproductive Strategies in Sexual Organisms: III. The Effects of Lottery Density Dependence and Pollen Limitation AB - This paper extends our previous work on modelling, within a single framework, the allocation of resources to reproduction vs. survival and the male vs. female components of reproduction in perennial plants. We derive the evolutionarily stable strategy (ESS) results under pollen limitation for both hermaphroditic and dioecious plant populations held stable through density-dependent juvenile recruitment. Pollen limitation affects female reproductive allocation in our model because there is post-flowering provisioning of offspring. We find that pollen limitation is unimportant to the ESS reproductive allocation and sex allocation so long as there are enough seeds to fill the empty sites left by the death of adults. To the extent that the relationships between gamete output and resource investment are linear for both sexes or sex functions, the separate treatment of reproductive and sex allocation in modern life-history and sex allocation theories is adequate. In such cases, the ESS sex allocation is exactly what is found in traditional sex allocation theory, and the ESS reproductive allocation of hermaphrodites or females in a dioecious species maximizes the amount of resources allocated to reproduction during an average lifespan, an analogue of the usual maximization principle in life-history theory modified to include the possibility of pollen limitation and extended seed maturation. The ESS reproductive allocation of males in a dioecious species maximizes lifetime pollen production, independent of pollen limitation and the female's resource allocation. Copyright 1997 Academic Press Limited PMID- 9344724 TI - A Reaction-Diffusion Model can Account for the Anatomical Pattern of the Cardiac Conal Valves in Fish AB - The conus arteriosus of fish bears a variable number of swallownest-like valves arranged in transverse rows. We propose that a reaction-diffusion mechanism coupled with the specific growth dynamics of the conus arteriosus, and probably, with a system of positional information, can account for the pattern formation of the conal valves in these primitive vertebrates. The reaction-diffusion mechanism, according to our hypothesis, would determine the regular arrangement of the places where the endocardial cells are activated and they transform into mesenchymal cells. These cells populate the subendocardial space, giving rise to the primordia of the conal valves. We have carried out simple computer simulations, based on reaction-diffusion/lateral inhibition models, which were able to generate all the anatomical diversity of the fish conal valves allowing changes only in the growth rate of the structure. Furthermore, we have found similarities between the sequential morphologies generated by our simulations and those actually occurring in embryos of Scyliorhinus canicula. We have derived three testable predictions from our hypothesis: (1) the number of discrete areas of epithelial-mesenchymal transition should correlate with the definitive number of valves in the conus arteriosus; (2) the small valves intercalated between the large ones should develop later than these latter; and (3) the longitudinal growth rate of the conus arteriosus in fish should be proportional to the number of transverse rows of valves. Copyright 1997 Academic Press Limited PMID- 9344725 TI - Doing What Everybody Does? A Procedure for Investigating Behavioural Synchronization AB - Behavioural synchronization means that the behaviour of several individuals is related in time. They may show the same behaviour either at the same time (synchrony) or explicitly at different moments (anachrony). Here, a procedure is presented, which not only allows quantitative investigation of behavioural synchronization in groups of any size, but also a separate evaluation with respect to each individual and behaviour pattern. The method distinguishes between the purely descriptive degree of concurrence and the degree of synchrony, which takes into account synchrony or anachrony occurring by chance on the basis of particular behaviour frequencies. Some recommendations for the application of the procedure are given and its limitations discussed. Copyright 1997 Academic Press Limited PMID- 9344726 TI - Dynamics of Forest Insect Density: Bifurcation Approach AB - Six basic phase portraits differing in character and in the number of their equilibrium behaviour regimes were suggested in the classification of forest insect population dynamics. The portraits graphically illustrate six types of dynamic behaviour of numbers of the system "phytophage-entomophage": two types of stable equilibrium dynamics (stationary and oscillatory) and four types corresponding to outbreaks of mass reproduction-fixed, permanent, reversive and true outbreaks, respectively. In this paper we consider the single parameter model, with its parametric domains realizing a wide spectrum of phase dynamics, both basic and transitional. The changes in the phase portraits are accompanied by the bifurcations in the model. The results of these analyses are in agreement with the "stability principle of mobile ecological systems" and may supply its parameter substantiation at the model level. Copyright 1997 Academic Press Limited PMID- 9344727 TI - Chaos in Glycolysis AB - Glycolysis occurs in almost every living cell as part of the energy metabolism. It forms a complex dynamical system, and might thus be capable of exhibiting complex phenomena. Simple oscillations have been observed frequently in suspensions of intact cells and in cell extracts, but only as transients. We have obtained sustained simple and complex oscillations in glycolysis of cell-free yeast extract in a flow-reactor. Sustained oscillations enable a powerful, proven method of dynamical system theory to unravel the kinetics and make it possible to observe chaos. Chaos was predicted from models long ago but has not previously been observed experimentally. We report the first experimental observation of unforced chaotic oscillations in glycolysis. Copyright 1997 Academic Press Limited PMID- 9344728 TI - Modelling Rooting Depth and Soil Strength in a Drying Soil Profile AB - A combined root growth and water extraction model is described that simulates the affects of mechanical impedance on root elongation in soil. The model simulates the vertical redistribution of water in the soil profile, water uptake by plant roots, and the effects of decreasing water content on increasing soil strength and decreasing the root elongation rate. The modelling approach is quite general and can be applied to any soil for which a relation can be defined between root elongation and penetrometer resistance. By definition this excludes soils that contain a large proportion of continuous channels through which roots can grow unimpeded. Root elongation rate is calculated as a function of the penetrometer resistance which is determined by the soil water content. Use of the model is illustrated using input data for a sandy loam soil. The results confirm reports in the literature that the depth of water extraction can exceed the rooting depth. The increase in mechanical impedance to root growth due to this water extraction restricted the maximum rooting depth attained, and this limited the depth of soil from which a crop could extract water and nutrients. This study highlighted the lack of published data sets for single crop/soil combinations containing both the strength/root growth information and the hydraulic conductivity characteristics necessary for this type of model. Copyright 1997 Academic Press Limited PMID- 9344729 TI - Modeling effects of Photoadaption on the Photosynthesis-Irradiance Curve AB - In this article, I analyse how photoadaption affects the photosynthesis irradiance curve in phytoplankton. Four parameters are presumably affected by photoadaption: (1) the size of a photosynthetic unit; (2) the number of photosynthetic units per cell; (3) the average turnover time of a photosynthetic unit; and (4) the chlorophyll-specific absorption cross-section. Prezelin's well known conceptual model of photoadaption deals with variation in size and number of photosynthetic units, but not with the other two parameters. Prezelin's model predicts that the photosynthesis-irradiance curve is differentially affected by variations in size or number of photosynthetic units. By contrast, my analysis shows that if photoadaptional variation in the turnover time is also accounted for, only the cellular chlorophyll concentration is relevant; variation in the size vs. number of photosynthetic units is immaterial. The photoadaptional response of the chlorophyll-specific absorption cross-section allows a simple description consistent with experimental data. I use this description to derive a single expression for the rate of photosynthesis as dependent on irradiance and the cellular chlorophyll concentration. This model for the photosynthesis irradiance-curve accounts for the photoadaptional alterations of all four parameters. Copyright 1997 Academic Press Limited PMID- 9344730 TI - Mortality During Dispersal an the Stability of a Metapopulation AB - We use a coupled map lattice to investigate the dynamics of a system of populations linked by dispersal, when dispersal incurs an additional mortality cost. Considering a single isolated population first, we show analytically that imposing an additional mortality term can stabilise the non-trivial steady state only under certain conditions. We demonstrate algebraically that in the absence of mortality during dispersal, linking a number of similar populations does not affect whether or not the equilibrium will be stable, although it can affect the nature of any unstable dynamics. Adding a fixed mortality rate during dispersal has a strong stabilising effect on system dynamics. We show analytically that for any combination of intrinsic reproduction parameters, a range of dispersal rates and dispersal mortalities can be found which together act to stabilise the equilibrium. Our results are shown numerically to be robust against a number of perturbations. Hence dispersal mortality has a strong stabilising effect on dynamics. In natural systems, some losses during long-distance dispersal are likely, and so we suggest that this factor could be an important determinant of the strength of observed population fluctuations. Copyright 1997 Academic Press Limited PMID- 9344731 TI - "First step" negative feedback accounts for inhibition of fast neurotransmitter release. AB - This paper is concerned with feedback inhibition of neurotransmitter release by the neurotransmitter itself. We put forward the idea that, similar to multistep biochemical processes, feedback inhibition acts on the initial step in the chain of events that lead to release. Using experimental results carried out on glutamatergic synapses in crayfish, we show that the "first step" hypothesis can account for all experimental results. Our modeling suggests that the biochemical implementation of this inhibition involves the formation of a second messenger, whose production is triggered by binding of transmitter to the autoreceptor. We argue that the autoreceptor is a key part of the release-inducing machinery. PMID- 9344732 TI - Computer model: investigating role of filopodia-based steering in experimental neurite galvanotropism. AB - Since early in this century developing axons and dendrites in culture have been reported to grow along electric field lines. It is only in the last score of years, however, that evidence suggests developing neurites actually orient in response to the electrical stimulus. We are interested in how an imposed electric field appears to speed neurite outgrowth in a field-related direction. We ask the question whether enhanced outgrowth in one direction results from streamlining outgrowth in that direction or from differentially catalysing the rate of outgrowth. Evidence for possible mechanisms of such neurite galvanotropism includes an electric field-dependent redistribution of filopodia, the finger-like structures that extend from the growing neurite tip. Using simple rules based on filopodia-mediated substrate sampling and orientation of extending neurites in vitro, we have built a computer model to test the streamlining theory. This in silico model of non-branching neurite outgrowth in two dimensions possesses the capacity to apportion its sampling efforts relative to a fixed reference representing the orientation of the field lines of a steady uniform electric field. Our model suggests that simple outgrowth patterns observed for experimental neurite galvanotropism-deflected and enhanced neurite growth toward the negative electrode and reduced neurite growth directed toward the positive electrode-may be simulated by tipping the balance of filopodia in the direction of the negative electrode. The existence of an analogous pattern-generating interaction between an applied electric field and extending neuronal processes would suggest a role for endogenous fields arising from naturally occurring potential gradients in developing organisms. PMID- 9344733 TI - Dynamics of the emergence of genetic resistance to biocides among asexual and sexual organisms. AB - A stochastic, agent based, evolutionary algorithm, modeling mating, reproduction, genetic variation, phenotypic expression and selection was used to study the dynamic interactions affecting a multiple-gene system. The results suggest that strong irreversible constraints affect the evolution of resistance to biocides. Resistant genes evolve differently in asexual organisms compared with sexual ones in response to various patterns of biocide applications. Asexual populations (viruses and bacteria) are less likely to develop genetic resistance in response to multiple pesticides or if pesticides are used at low doses, whereas sexual populations (insects for example) are more likely to become resistant to pesticides if susceptibility to the pesticide relates to mate selection. The adaptation of genes not related to the emergence of resistance will affect the dynamics of the evolution of resistance. Increasing the number of pesticides reduces the probability of developing resistance to any of them in asexual organisms but much less so in sexual organisms. Sequential applications of toxins, were slightly less efficient in slowing emergence of resistance compared with simultaneous application of a mix in both sexual and asexual organisms. Targeting only one sex of the pest speeds the development of resistance. The findings are consistent to most of the published analytical models but are closer to known experimental results, showing that nonlinear, agent based simulation models are more powerful in explaining complex processes. PMID- 9344734 TI - General equation of steady-state enzyme kinetics using net rate constants and its applicaiton to the kinetic analysis of catalase reaction. AB - The steady-state velocity equation is derived for the general reaction scheme containing n kinds of enzyme species connected by a network of reversible reaction steps. The general equation is represented by the net rate constants as well as the true rate constants of the individual reaction steps. Using a general equation, equations for simpler schemes can easily be derived. Furthermore, the velocity equation expressed by net rate constants is useful for understanding the dynamic state of any reaction be it simple or complex. The generality of the presented equation is tested in a complex reaction involving Bacillus stearothermophilus catalase I. In addition, the applicability of the general equation in analysing the reaction specificity and dynamic state of the reaction is shown. PMID- 9344735 TI - Theoretical description of the polymerase chain reaction. AB - Taking into account expressions for the efficiency of the polymerase chain reaction (PCR) recently deduced from enzymological considerations, and making use of a continuous model based on the law of mass action, closed form solutions are derived that enable a complete description of the standard and quantitative competitive PCR methods. The resulting behaviour is in reasonable agreement with that from a previous, empirical, discrete approach; but the latter is nonetheless shown to overestimate the amplification yield by as much as 30% due to the weak assumption of a constant efficiency during the cycle duration. The present formalism will facilitate the implementation of accurate fitting procedures of experimental data to manage quantification. PMID- 9344738 TI - Parallel overlap assembly for the construction of computational DNA libraries. AB - Algorithms for computing with DNA currently require the construction of pools of molecules in which each distinct molecule represents a different starting point for the calculation. We have begun building such pools using the technique of parallel overlap assembly that is already used for the generation of diversity in biologically useful combinatorial search techniques such as gene shuffling. Unlike these applications, a pool in a molecular computer must be complete, containing all possible strands, and ordered, having minimal contamination from incorrectly assembled DNA. We present an experiment in which parallel overlap assembly is used to construct a computational pool and an experiment in which this pool is used to solve the NP-complete maximal-clique problem. PMID- 9344739 TI - The preferential mode analysis of DNA sequence. AB - After reviewing approaches to the nucleotide correlation of DNA sequences the preferential mode analysis method is emphasized and discussed in detail. The preferred modes and poor modes in coding regions, as well as in introns, 5'-caps and 3'-tails are found through the statistical analysis of sequence data of all kinds of species in GenBank. The relation between the preferential mode analysis and informational parameter method is deduced. It is discovered that in higher species the coding sequences preferentially use the strong-weak bond (strong bond=C,G; weak bond=A, T) language and many noncoding regions (introns, 5'-caps, 3'-tails) use purine-pyrimidine language. The application of different languages in coding and noncoding sequences is a result of evolution, and it may be related to the functional differences in these two regions. Furthermore, we find that many preferential triplets in coding sequences can be expressed in a form of (* W S) (W=A,T; S=C,G), which may be explained by its relation to t-RNA abundance. The systematic change of some mode contents with evolution has also been found. PMID- 9344740 TI - Improving the efficiency of the genetic code by varying the codon length--the perfect genetic code. AB - The function of DNA is to specify protein sequences. The four-base "alphabet" used in nucleic acids is translated to the 20 base alphabet of proteins (plus a stop signal) via the genetic code. The code is neither overlapping nor punctuated, but has mRNA sequences read in successive triplet codons until reaching a stop codon. The true genetic code uses three bases for every amino acid. The efficiency of the genetic code can be significantly increased if the requirement for a fixed codon length is dropped so that the more common amino acids have shorter codon lengths and rare amino acids have longer codon lengths. More efficient codes can be derived using the Shannon-Fano and Huffman coding algorithms. The compression achieved using a Huffman code cannot be improved upon. I have used these algorithms to derive efficient codes for representing protein sequences using both two and four bases. The length of DNA required to specify the complete set of protein sequences could be significantly shorter if transcription used a variable codon length. The restriction to a fixed codon length of three bases means that it takes 42% more DNA than the minimum necessary, and the genetic code is 70% efficient. One can think of many reasons why this maximally efficient code has not evolved: there is very little redundancy so almost any mutation causes an amino acid change. Many mutations will be potentially lethal frame-shift mutations, if the mutation leads to a change in codon length. It would be more difficult for the machinery of transcription to cope with a variable codon length. Nevertheless, in the strict and narrow sense of coding for protein sequences using the minimum length of DNA possible, the Huffman code derived here is perfect. PMID- 9344742 TI - Estimating the entropy of DNA sequences. AB - The Shannon entropy is a standard measure for the order state of symbol sequences, such as, for example, DNA sequences. In order to incorporate correlations between symbols, the entropy of n-mers (consecutive strands of n symbols) has to be determined. Here, an assay is presented to estimate such higher order entropies (block entropies) for DNA sequences when the actual number of observations is small compared with the number of possible outcomes. The n-mer probability distribution underlying the dynamical process is reconstructed using elementary statistical principles: The theorem of asymptotic equi-distribution and the Maximum Entropy Principle. Constraints are set to force the constructed distributions to adopt features which are characteristic for the real probability distribution. From the many solutions compatible with these constraints the one with the highest entropy is the most likely one according to the Maximum Entropy Principle. An algorithm performing this procedure is expounded. It is tested by applying it to various DNA model sequences whose exact entropies are known. Finally, results for a real DNA sequence, the complete genome of the Epstein Barr virus, are presented and compared with those of other information carriers (texts, computer source code, music). It seems as if DNA sequences possess much more freedom in the combination of the symbols of their alphabet than written language or computer source codes. PMID- 9344743 TI - Nucleosomes: a solution to a crowded intracellular environment? AB - The emergence of eukaryotes was accompanied by two major events that concern their genome and are of crucial significance when considered in terms of macromolecular crowding: (i) a substantial increase in the amount of DNA, and (ii) its confinement within a defined space. The resulting highly crowded environment would have strongly promoted DNA self-assembly processes, leading to extremely condensed and thermodynamically stable DNA aggregates. Such structural transitions have indeed been observed in vitro, as well as in virtually all cellular systems in which a nucleosomal assembly is absent. In this appear we raise the hypothesis that upon transition from prokaryotic systems to eukaryotes, the nucleosomes were rendered essential in order to negate extensive DNA condensation processes that would have resulted from excluded volume effects. By suppressing such processes, the nucleosomes act to maintain and regulate the conformational space of the DNA, thus enabling conformational flexibility and reversible structural modulations. PMID- 9344744 TI - Kinetic analysis of chemical or enzymic reactions: an algorithm for the determination of the initial velocity of product formation by the use of a taylor series in reaction time. PMID- 9344745 TI - Validation of protein models from Calpha coordinates alone. AB - The Protein Data Bank contains a number of structures for which only the coordinates of the Calpha atoms have been deposited. Although many tools are available for the validation of all-atom protein models, hardly any of these can be used to assess the quality of models for which only Calpha coordinates are available. Two rapid and simple tests to assess the quality of the Calpha backbone of a protein model are described, one based on the distribution of Calpha-Calpha distances, and the other on the two-dimensional distribution of the angles and dihedrals formed by sequential Calpha atoms. Expected distributions were derived by analysing a set of 1343 high-resolution, all-atom protein models. The distance criterion is useful to discriminate between refined and unrefined models, whereas the angle/dihedral criterion can be used to discriminate between normal and possibly problematic Calpha models. The method has been applied to a set of 88 Calpha-only models from the Protein Data Bank. The tracing of two of the models that are outliers in this analysis has recently been shown to be incorrect. Other applications of the method are discussed. PMID- 9344746 TI - Role of upstream activation sequences and integration host factor in transcriptional activation by the constitutively active prokaryotic enhancer binding protein PspF. AB - PspF, the transcriptional activator of the pspA operon of Escherichia coli, which belongs to the enhancer binding protein (EBP) family of sigma54 activator proteins, is constitutively active in an in vitro transcription assay. PspF protein, together with RNA polymerase holoenzyme containing sigma54, is required for in vitro transcription from the pspA promoter. EBP proteins are typically subject to regulation either by post-translational modification or interaction of a specific ligand with an N-terminal regulatory domain. However, unlike other members of the EBP family, PspF lacks this domain. pspA is positively regulated by IHF in vitro, and this regulation is dependent on the topology of the DNA; a linear template is much more dependent on IHF than a supercoiled template. EBP binding to upstream activating sequences (UAS) in their target promoters is mediated by the C-terminal domain which contains a helix-turn-helix DNA-binding motif. A mutant PspF protein lacking the C-terminal DNA-binding domain is active in vitro, although at much higher concentrations than the wild-type protein. In vitro transcription from pspA templates missing one or both of the UAS sites is reduced relative to wild-type templates, but is still appreciable; however, IHF acts as a negative regulator of pspA transcription on these mutant templates. Thus, PspF bound to non-specific sequences upstream of the pspA promoter can activate pspA transcription, but this activation is inhibited by IHF. These data, taken together, support the model that a precise promoter geometry is necessary for IHF to positively regulate transcription and that IHF may act to prevent activation from inappropriately spaced upstream sites. PMID- 9344747 TI - Release factor RF3 abolishes competition between release factor RF1 and ribosome recycling factor (RRF) for a ribosome binding site. AB - The dependence of the rate of ribosomal recycling (from initiation via protein elongation and termination, and then back to initiation) on the concentrations of release factor RF1 and the ribosome recycling factor (RRF) has been studied in vitro. High RF1 concentration was found to reduce the rate of ribosomal recycling and the extent of this reduction depended on stop codon context. The inhibitory effect of high RF1 concentrations can be reversed by a corresponding increase in RRF concentration. This indicates that RF1 and RRF have mutually exclusive and perhaps overlapping binding sites on the ribosome. Addition of release factor RF3 to the translation system abolishes the inhibitory effect of high RF1 concentration and increases the overall rate of ribosome recycling. These data can be explained by a three-step model for termination where the first step is RF1-promoted hydrolysis of peptidyl-tRNA. The second step is an intrinsically slow dissociation of RF1 which is accelerated by RF3. The third step, catalysed by RRF and elongation factor G, leads to mobility of the ribosome on mRNA allowing it to enter a further round of translation. In the absence of RF3, RF1 can re-associate rapidly with the ribosome after peptidyl-tRNA hydrolysis, preventing RRF from entering the ribosomal A-site and thereby inhibiting ribosomal recycling. The overproduction of RF1 in cells deficient in RRF or lacking RF3 has effects on growth rate predicted by the in vitro experiments. PMID- 9344748 TI - A folded monomeric intermediate in the formation of lambda Cro dimer-DNA complexes. AB - The folding, dimerization and DNA binding equilibria of the bacteriophage lambda Cro repressor have been characterized. Comparison with four engineered variants shows that a folded monomeric species is substantially populated under conditions used for the formation of dimer-DNA complexes. Although Cro dimers are the only DNA-bound species observed in electrophoretic mobility shift assays, cooperativity in Cro-DNA binding isotherms shows that the predominant free protein species is monomeric at nanomolar concentrations. Micromolar dissociation constants for Cro dimers have been measured in the absence of DNA by sedimentation equilibrium and gel filtration chromatography. Denaturation of Cro dimers in the 10 to 100 micromolar concentration range by guanidine hydrochloride (GdnHCl) is well modeled as a two-state process, with folded dimers and unfolded monomers as the only significantly populated species. However, linear extrapolation of this composite unfolding and dimer dissociation free energy predicts a nanomolar dissociation constant in the absence of denaturant. This extrapolation is clearly inconsistent with the DNA binding and hydrodynamic measurements. Our interpretation of these results is that the monomeric species detected in DNA binding and hydrodynamic experiments is predominantly folded. The stability of the folded monomeric species can be calculated as the difference between the dimerization free energy determined from hydrodynamic measurements and the folding free energy extrapolated from GdnHCl denaturation. The calculated stability of the Cro F58W monomer is greater than that of the wild-type Cro monomer. Thus, residue 58, which makes critical intermolecular contacts across the dimer interface, is also involved in intramolecular stabilization of the monomeric intermediate. PMID- 9344749 TI - DNA repair of UV photoproducts and mutagenesis in human mitochondrial DNA. AB - The induction and repair of DNA photolesions and mutations in the mitochondrial (mt) DNA of human cells in culture were analysed after cell exposure to UV-C light. The level of induction of cyclobutane pyrimidine dimers (CPD) in mitochondrial and nuclear DNA was comparable, while a higher frequency of pyrimidine (6-4) pyrimidone photoproducts (6-4 PP) was detected in mitochondrial than in nuclear DNA. Besides the known defect in CPD removal, mitochondria were shown to be deficient also in the excision of 6-4 PP. The effects of repair defective conditions for the two major UV photolesions on mutagenesis was assessed by analysing the frequency and spectrum of spontaneous and UV-induced mutations by restriction site mutation (RSM) method in a restriction endonuclease site, NciI (5'CCCGG3') located within the coding sequence of the mitochondrial gene for tRNALeu. The spontaneous mutation frequency and spectrum at the NciI site of mitochondrial DNA was very similar to the RSM background mutation frequency (approximately 10(-5)) and type (predominantly GC>AT transitions at G1 of the NciI site). Conversely, an approximately tenfold increase over background mutation frequency was recorded after cell exposure to 20 J/m2. In this case, the majority of mutations were C>T transitions preferentially located on the non transcribed DNA strand at C1 and C2 of the NciI site. This mutation spectrum is expected by UV mutagenesis. This is the first evidence of induction of mutations in mitochondrial DNA by treatment of human cells with a carcinogen. PMID- 9344750 TI - Residues of the cytoplasmic domain of MotA essential for torque generation in the bacterial flagellar motor. AB - The MotA protein of Escherichia coli is a component of the flagellum that functions, together with MotB, in transmembrane proton conduction. MotA and MotB are believed to form the stator of the flagellar motor. They are integral membrane proteins; MotA has a large (ca 22 kDa) domain in the cytoplasm, and MotB a much smaller one (ca 3 kDa). Recent work suggests that cytoplasmically located parts of MotA and/or MotB might be present at the active site for torque generation in the motor. To test the proposal that the cytoplasmic domain of MotA functions in torque generation, and to identify the amino acid residues most important for function, we have carried out a mutational analysis of this domain. Using random mutagenesis, many mutations of cytoplasmic residues of MotA were isolated, which either abolish or impair torque generation. In most cases the residues affected are not conserved, and many of the replacements involve loss or gain of a proline residue, which suggests that these mutations disrupt function by altering the protein conformation rather than by directly affecting residues of an active site. Using site-directed mutagenesis, the conserved residues in the cytoplasmic domain of MotA were replaced, either singly or, in the case of charged residues, in various combinations. The results identify four residues of MotA that are important for motor function. These are Arg90 and Glu98, located in the cytoplasmic domain, and Pro173 and Pro222, located at the interface between the cytoplasmic domain and the membrane-spanning domain. Possible roles for these residues in torque generation are discussed. PMID- 9344751 TI - Myosin head configuration in relaxed fish muscle: resting state myosin heads must swing axially by up to 150 A or turn upside down to reach rigor. AB - The arrangement and shape of myosin heads in relaxed muscle have been determined by analysis of low-angle X-ray diffraction data from a very highly ordered vertebrate muscle in bony fish. This reveals the arrangement and interactions between the two heads of the same myosin molecule, the shape of the resting myosin head (M.ADP.Pi) assuming a putative hinge between the myosin catalytic domain and the light chain binding-domain, and the way that the actin-binding sites on myosin are arrayed around the actin filaments in the bony fish muscle A band cell unit. The results are discussed in terms of possible force-generating mechanisms. Changes in myosin head shape or tilt have been implicated in the mechanism of force generation. The myosin head arrangement, including perturbations from perfect helical symmetry, has all heads oriented roughly the same way up (there is only a small range of rotations around the head long axis). X-ray data do not define the absolute polarity of the myosin head array. The resting head rotation is either similar to (65 degrees difference) or opposite to (115 degrees difference) the rotation in the rigor state. If the rotations are similar, probably the more likely possibility, then the average relative axial displacement of the inner and outer ends of the heads from the resting state to rigor is about 140 to 150 A. If (less likely) the resting head rotation is opposite to rigor, then the heads would need to turn over (i.e. rotate about 115 degrees around their own long axes) and the mean relative axial displacement from relaxed to rigor would only be 20 to 30 A. PMID- 9344752 TI - Partially folded states of the capsid protein of cowpea severe mosaic virus in the disassembly pathway. AB - The different partially folded states of the capsid protein that appear in the disassembly pathway of cowpea severe mosaic virus (CPSMV) were investigated by examining the effects of hydrostatic pressure, sub-zero temperatures and urea. The conformational states of the coat protein were analyzed by their intrinsic fluorescence, binding of bis(8-anilinonaphthalene-1-sulfonate) (bis-ANS) and susceptibility to trypsin digestion. CPSMV could be disassembled by pressure at 2.5 kbar. Intrinsic fluorescence and hydrodynamic measurements showed that pressure-induced dissociation was completely reversible. Virus pressurization in the presence of ribonuclease revealed that viral RNA was not exposed, since it was not digested by the enzyme, suggesting the maintenance of protein-nucleic acid interactions under pressure. When the temperature was decreased to -10 degrees C under pressure, CPSMV disassembly became an irreversible process and in this condition, viral RNA was completely digested by ribonuclease. These results suggest a relationship between protein-RNA interactions and CPSMV assembly. Bis ANS binding and trypsin digestion of coat proteins revealed that they assume a different conformation when they are denatured by low temperatures under pressure or than when they are denatured by urea at atmospheric pressure. The results indicate that the coat proteins can exist in at least four states: (1) The native conformation in the virus capsid; (2) bound to RNA when the virus is dissociated by pressure at room temperature, assuming a conformation that retains the information for reassembly; (3) free subunits in a molten-globule conformation when the virus is dissociated by low temperature under pressure; and (4) free subunits completely unfolded by high concentrations of urea. PMID- 9344753 TI - Mutant ATP-binding RNA aptamers reveal the structural basis for ligand binding. AB - The solution structure of the ATP-binding RNA aptamer has recently been determined by NMR spectroscopy. The three-dimensional fold of the molecule is determined to a large extent by stacking and hydrogen bond interactions. In the course of the structure determination it was discovered that several highly conserved nucleotides in the binding pocket can be substituted while retaining binding under NMR conditions. These surprising findings allow a closer look at the interactions that determine stability and specificity of the aptamer as well as local structural features of the molecule. The binding properties of ATP binder mutants and modified ligand molecules are explored using NMR spectroscopy, column binding studies and molecular modeling. We present additional evidence and new insights regarding the network of hydrogen bonds that defines the structure and determines stability and specificity of the aptamer. PMID- 9344754 TI - Charges, hydrogen bonds, and correlated motions in the 1 A resolution refined structure of the mating pheromone Er-1 from Euplotes raikovi. AB - A detailed description is given of the structure of the small protein mating pheromone Er-1 at atomic resolution. Emphasis is placed on the locations of charges and hydrogen bonds. The model includes all the protein atoms, anisotropic displacement parameters, four disordered side chains, 22 water molecules, a disordered ethanol, and "riding" hydrogen atoms. Analysis of the model revealed that this dense crystal is perfused by hydrogen-bonding networks of solvent and protein atoms. The termini of helices are capped by hydrogen bonding to solvent and protein atoms, and to symmetry-related molecules. An examination of the valencies and charges of the hydrogen-bonding groups suggests that three of the "water" molecules capping the C termini of two helices, and one other, may instead be NH4 ions. Water molecules mediate all but one of the interhelical hydrogen bonds, and many of the lattice interactions. Regions of the molecule where the atomic vibrations deviate from isotropy are identified. There is almost no overall libration of the molecule allowed by the packing, but the side-chains vibrate relative to the backbone. Four side-chains display alternate conformations. Indirect evidence is presented that the switches between their conformations are correlated and driven by protonation of acidic side-chains. These structural features are discussed in the context of function and stability. Equipped with the analysis of the model, we review the course and results of the refinement of the model against 1 A X-ray diffraction data to a crystallographic R-factor of 12.92%. PMID- 9344755 TI - Nitric oxide attenuates reoxygenation-induced ICAM-1 expression in coronary microvascular endothelium: role of NFkappaB. AB - Enhanced leukocyte adhesion has been shown to occur in post-ischemic reperfused hearts due to the upregulation of specific cell-surface adhesion molecules. Therefore, we investigated the influence of 4 h of reoxygenation after 20 h of hypoxia on ICAM-1 induction in primary cultures of rat coronary microvascular endothelial cells (CMEC). ICAM-1 surface expression as well as oxygen free radical formation were measured by flow cytometry. Changes in ICAM-1 mRNA levels were assessed by Northern blot and activation of NFkappaB and AP-1 signalling were analysed by electrophoretic mobility shift assays (EMSA) in CMEC lysates. Although hypoxia alone did not affect cell-surface ICAM-1 expression, 4 h of reoxygenation induced a significant upregulation of ICAM-1. ICAM-1 mRNA could not be found after hypoxia alone, but could be detected as early as 1 h following reoxygenation. Unlike AP-1, the activation of which could be detected in CMEC lysates following hypoxia alone, NFkappaB binding activity was induced only following reoxygenation, concurrent with an increase in the formation of reactive oxygen species (ROS). A proteasome inhibitor, nor-Leu (25 microM) inhibited NFkappaB activation by reoxygenation and ICAM-1 expression. Blockade of endogenous nitric oxide (NO) synthesis in CMEC with L-nitroarginine (10 microM) accentuated post-reoxygenation ICAM-1 expression. Finally, an exogenous NO donor, S-nitrosoacetyl-penicillamine (SNAP, 100 microM), suppressed the generation of ROS upon reoxygenation, and blocked the activation of NFkappaB and the upregulation of ICAM-1. Thus, ICAM-1 upregulation in CMEC primary cultures is not induced by hypoxia alone, but appears shortly after reoxygenation in the absence of exogenous cytokines or inflammatory cells. Because upregulation of AP-1 through hypoxia alone did not affect ICAM-1 expression, we conclude that redox sensitive NFkappaB activation triggers ICAM-1 upregulation. NO inhibits reoxygenation-specific ICAM-1 upregulation, most likely by diminishing oxidative stress that leads to NFkappaB activation. PMID- 9344756 TI - The changing metabolic role of fatty acids in increasingly complex organisms: an evolutionary perspective. PMID- 9344758 TI - Contractile failure in chronic doxorubicin-induced cardiomyopathy. AB - The mechanisms for the progression of the anthracycline-induced cardiomyopathy to contractile failure has not been defined. In vitro, doxorubicin (DOX) appears to modify calcium-mediated excitation-contraction coupling, which depresses cardiac contractility. This study characterizes the onset of contractile failure associated with the development of DOX-induced cardiomyopathy. Rabbits were treated with DOX (1 mg/kg i.v. twice weekly, 12-18 doses; DOX-treated group) and compared with a pair-fed Control group infuse with saline vehicle. The severity of the cardiomyopathy was determined by numerically-scored histopathology. Myocardial contractility was determined in thin fiber bundles from right ventricular (RV) papillary muscles and left atria that were removed and mounted on a force transducer in oxygenated Krebs-bicarbonate buffer (pH=7.4 at 30 degrees C) to record the amplitude (DT) and maximum rate (+dT/dt ) of isometric tension. Myofibrillar and calcium loading properties were determined by the calcium and caffeine-activated tension responses respectively in chemically permeabilized fibers. With the onset of the cardiomyopathy (score <2) DT at low frequency (0.5 Hz) was depressed (0.61+/-0.01 mN/mg; n=14) compared to Control (0.93+/-0.09 mN/mg; n=15). Contractility at higher rates (1 Hz) was not different in this DOX-treated and Control groups. Maximum calcium and caffeine-activated force and the pCa to half-maximum force of permeabilized fibers were comparable in DOX-treated and Control groups. The loss of contractility of the DOX-treated group was related to reduction in sarcoplasmic reticulum calcium release channel density, as determined by Bmax for 3H-ryanodine binding in cardiac microsomal membrane fraction. Post-rest potentiation of contractility, as well as frequency dependent (0.25-1.5 Hz) and post-extrasystolic potentiation of contractility were preserved in the DOX-treated group. In vitro, DOX depressed post-rest potentiation of contractility. Thus, the onset of contractile failure of the DOX induced cardiomyopathy is characterized by effects consistent with disordered calcium-mediated excitation-contraction coupling and these effects are qualitatively different than in vitro effects of DOX. PMID- 9344759 TI - Myocardial infarction and regulatory myosin light chain. AB - Myosin from cardiac muscle consists of two heavy chains and two pairs of light chain. Regulatory myosin light chain (RMLC) is phosphorylated by a Ca2+ and calmodulin dependent myosin light chain kinase. The impact of experimental myocardial infarction on cardiac RMLC was studied. The left anterior descending coronary artery of rabbits was ligated. Three, 7 and 14 days later the animals were euthanized, sections of the heart were frozen in liquid nitrogen and later subjected to 2-dimensional electrophoresis. Isoelectric focusing was carried out at a pH range of 4.5-5.4. Reproducible patterns of protein separation showed four spots with proteins of phosphorylatable regulatory light chains shifted to a more negative pH as compared to essential light chain. We investigated changes in phosphorylation of RMLC in infarcted heart muscle. As compared to sham operated animals, a decline in phosphorylation of RMLC was present in both infarcted and non-infarcted portions of the left ventricle; the latter was significant 7 days following the onset of ischemia. In contrast, the decline in percent phosphorylation in the infarcted area was not significant. The amount of RMLC decreased significantly in the infarcted portion. A highly significant reduction in the percent of viable cardiomyocytes accompanied the decline in phosphorylation. There was a significant correlation of RMLC following administration of isoproterenol, 7 and 14 days following onset of ischemia. Only faint traces of essential atrial myosin light chain (ALC-1) were present in the non-infarcted portion of the left ventricle. No correlation was found between percent phosphorylation and the amount of RMLC (density) following infusion of saline or isoproterenol. Isoproterenol significantly increased percent phosphorylation without altering the amount of RMLC protein. We conclude that myocardial infarction profoundly affects regulatory myosin light chain phosphorylation in the infarcted and non-infarcted areas of the myocardium and that RMLC plays a significant part in myocardial contractility. PMID- 9344757 TI - Subcellular distribution of ankyrin in developing rabbit heart--relationship to the Na+-Ca2+ exchanger. AB - Ankyrins are a multigene family of proteins that function as adapters between the cytoskeleton and trans-membrane proteins, such as ion channels. Previous studies have shown the linkage between ankyrin and ionic transport proteins such as Na+ K+ ATPase, voltage-dependent Na+ channels and Ca2+ channels. In the present study, we have investigated the subcellular distribution of ankyrin and its relationship to the Na+-Ca2+ exchange protein in immature and adult rabbit ventricular myocytes. Isolated single cardiomyocytes from neonatal, juvenile and adult rabbit hearts were examined by immunofluorescence labeling techniques, using antibodies against ankyrin and the Na+-Ca2+ exchanger. We found that in neonatal rabbit cardiac myocytes, ankyrin labeling was mainly present at the Z disk, whereas the Na+-Ca2+ exchanger was only present on the peripheral sarcolemma. At 2 weeks of age, ankyrin labeling was still predominantly observed at the level of the Z disks as well as in the partially developed T-tubules. In the adult cells, however, ankyrin and the Na+-Ca2+ exchanger seem to be co localized within T-tubules and at the costamere region of the peripheral sarcolemma. Immunogold labeling studies at the higher resolution electron microscopic level using cyrosection tissues of rabbit heart at different ages confirm these findings. These results indicate that the distribution pattern of ankyrin and the Na+-Ca2+ exchanger changes with development in rabbit ventricular myocytes. PMID- 9344760 TI - Changes in thyroid state affect pHi and Nai+ homeostasis in rat ventricular myocytes. AB - We have tested the hypothesis that thyroid state may influence both the flow of cellular Ca2+ and the myofilament response to Ca2+ by effects on intracellular pH (pHi) and Na+ (Nai+). Single cardiac myocytes isolated from hypothyroid, euthyroid and hyperthyroid animals were loaded with fura-2/AM (Cai2+ probe), BCECF/AM (pHi probe) or SBFI/AM (Nai+ probe). Compared with hypothyroid animals, myocytes isolated from hyperthyroid rat hearts demonstrated a significant: (1) increase in extent of shortening; (2) decrease in the time to peak contraction; (3) increase in the peak amplitude of the fura-2 fluorescence ratio; (4) decrease in pHi (DeltapHi=0. 19+/-0.05); and (5) increase in Nai+ (DeltaNai+=2.88+/-0.55 mM). We have also compared pHi in Langendorff perfused hypo- and hyperthyroid rat hearts using NMR. We have found that hyperthyroid hearts are 0.15+/-0.03 pH units more acidic than hypothyroid hearts. Analysis of mRNA levels demonstrated that hyperthyroidism increased expression of both the Na+/Ca2+ exchanger and Na+/H+ antiporter, and decreased expression of Na+ channel mRNAs. These changes appear partially responsible for the observed changes in Nai+ and pHi. Our results provide the first evidence that changes in cardiac contractility associated with altered thyroid state not only involve effects on Ca2+, but may also involve changes in the response of the myofilaments to Cai2+mediated by altered pHi and Nai+. PMID- 9344761 TI - Selective induction of the creatine kinase-B gene in chronic volume overload hypertrophy is not affected by ACE-inhibitor therapy. AB - Hypertrophied and failing myocardium has been shown to undergo creatine kinase (CK) isoform switching, resulting in increased MB and BB components. We tested the hypothesis that chronic volume overload hypertrophy due to mitral regurgitation in the dog causes CK isoenzyme switching and that this could be reversed by angiotensin converting enzyme inhibitor therapy. Thirteen adult mongrel dogs had mitral regurgitation induced by mitral valvular chordal rupture: six were treated with ramipril for 4 months and seven were untreated for 4 months. Twelve dogs were sham-operated: six received ramipril for 3 months and six were untreated. Left ventricular end-diastolic volume increased from 58+/-4 to 104+/-10 ml in untreated (P<0.001) and from 55+/-3 to 91+/-6 ml in treated dogs (P<0.01) as LV mass/volume ratio decreased in both untreated (1. 60+/-0.07 to 1.13+/-0.08 g/ml, P<0.001) and treated dogs (1.44+/-0.06 to 1.20+/-0.08 g/ml, P<0.01). CK-MB isoform was 7.4+/-1.1% in normal shams and increased to 13.5+/ 1.9% and 18.1+/-3.0% in both treated and untreated mitral regurgitation dogs; respectively (P<0. 05). Steady state CK-B mRNA increased three-fold in treated and untreated dogs with mitral regurgitation (P<0.003) compared to normals, while CK-M mRNA expression did not differ in all groups. Thus, chronic volume overload hypertrophy of mitral regurgitation induces CK isoform switching by selective induction of the CK-B gene, and ramipril therapy does not affect this isoform switch. This may reflect a response to increased diastolic stress and more efficient energy utilization in the volume overloaded myocardium. PMID- 9344762 TI - Neural crest is involved in development of abnormal myocardial function. AB - Around 85% of embryos homozygous for the splotch (Sp2H) allele (Sp2H/Sp2H), a Pax3 mutation, develop persistent truncus arteriosus (PTA), a defect related to the cardiac neural crest. These embryos die by 14.5 days post coitum. In an investigation of the cause of lethality in these embryos, we used digital video imaging microscopy to examine beating embryonic hearts in situ at 13.5 dpc. The hearts of Sp2H/Sp2H embryos with PTA clearly showed poor function when compared with normal litter mates. Contractile force was examined in detergent-skinned ventricular muscle strips from Sp2H/Sp2H embryos at ages 12.5 and 13.5 dpc. There was no significant difference in the maximum force or in myosin content between Sp2H/Sp2H and control groups, indicating no significant dysfunction of the contractile apparatus in hearts from Sp2H/Sp2H embryos. Ca2+ transients were examined in enzymatically-dissociated ventricular myocytes and were significantly reduced in defective hearts, indicating that reduced cardiac function in Sp2H/Sp2H embryos with PTA was due to impaired excitation-contraction (EC) coupling. Ca2+ currents were examined using the perforated patch clamp technique. The magnitude of the Ca2+ current was found to be reduced by approximately 3.2 fold in Sp2H/Sp2H hearts with PTA compared to normal. Since the sarcoplasmic reticulum is sparse or absent in the embryonic heart, the impaired EC coupling was due to the reduction in Ca2+ current. These observations suggest that neural crest abnormalities result in a defect in EC coupling, causing depressed myocardial function and death in utero from cardiac failure. Interestingly, Sp2H/Sp2H hearts without PTA had normal EC coupling. These results indicated that impaired EC coupling was secondary to the Pax3 mutation. The findings in this report indicate an important role for the neural crest in the development of normal myocardial function, and represent the first demonstration of impaired excitation-contraction coupling in a genetically-defined embryonic mammalian model of a cardiac structural defect. PMID- 9344763 TI - Basic FGF enhances calcium permeable channel openings in adult rat cardiac myocytes: implication in the bFGF-induced increase of free Ca2+ content. AB - Basic fibroblast growth factor (bFGF) has been implicated in the changes in gene expression that, under pathological conditions such as ischemia or volume overload, lead to adult cardiomyocyte hypertrophy. In many tissues, one of the first events following cell activation by growth factors is an enhancement of the intracellular free calcium concentration, generated by fluxes from internal storage compartments and through channels of the plasma membrane. The present study was undertaken to determine whether cardiac myocytes isolated from adult rat ventricles express Ca2+-permeable channels activated by bFGF. Using the cell attached mode of the patch-clamp technique, we observed that bFGF (from 0.1-10 nM) induced an increase of fast burst openings, mediated by Ca2+-permeable channels with low conductance (15 pS) and voltage-independence. Inside-out patch clamp experiments revealed that inositol 1,4,5-trisphosphate (5 microM) enhanced the opening of Ca2+-permeable channels with similar properties as the bFGF induced channels, indicating that IP3 may be a second messenger of this process. Confocal fluorescence imaging of intracellular free calcium provided direct evidence that bFGF induced an increase of cytoplasmic and nucleoplasmic free Ca2+ concentrations which were generated, in part, by Ca2+ influx through the plasma membrane. In conclusion, this study supports the presence, in the plasma membrane of adult cardiac myocytes, of messenger-activated calcium channels which could play key roles in the calcium-dependent pathways that are activated in response to growth factors. PMID- 9344764 TI - Point mutations in mitochondrial DNA of patients with dilated cardiomyopathy. AB - Mitochondrial (mt) DNA mutations are hypothesized to be involved in the pathogenesis of dilated cardiomyopathy, because the mtDNA encodes 13 polypeptides that are essential for oxidative phosphorylation, upon which the heart relies for energy. To test this hypothesis, we amplified the mitochondrial genome by long PCR and then used restriction analysis and direct sequencing to examine 58 unrelated patients with dilated cardiomyopathy and 49 controls for the detection of point mutations. The results demonstrated that point mutations were significantly more frequent in the mtDNA of patients than in that of controls (173 in 58 patients v 54 in 49 controls, 2=16.51, P<0.001). In addition to normal variants and mutations common to both patients and controls, 43 mutations were identified only in patients. All but four of these mutations were homoplasmic. Mutations involving the evolutionarily conserved residues of cytochrome c oxidase subunit I, NADH dehydrogenase 5, tRNAAla and tRNAArg were identified. As many as 13 point mutations were found in an 8-month-old patient. In conclusion, there exist significantly more point mutations in mtDNA of patients than in controls, suggesting that multiple mutations may exert a cumulative effect on heart function. Thus, by altering the function of respiratory enzyme subunits or tRNAs, mtDNA point mutations could be relevant for the pathogenesis of dilated cardiomyopathy. PMID- 9344766 TI - Telomerase activity during cardiac development. AB - Telomerase is a ribonucleoprotein involved in maintaining telomere length in stem cells and immortal and actively dividing cells. We report here for the first time that telomerase is developmentally regulated in the normal rat heart. When we compared rat hearts at different developmental stages, we found that telomerase activity decreased to 20% of the fetal level by 5 days after birth, and was undetectable by 20 days after birth. These results indicate that the rate of cardiomyocyte proliferation decreases dramatically soon after birth in the rat. Of several non-cardiac tissues examined, telomerase activity was highest in fetal and adult rat liver, suggesting that there is an active mechanism for maintaining long telomeres in liver tissue at all stages. The disappearance of telomerase activity in the rat heart at the time that cardiomyocytes become terminally differentiated suggests that telomerase downregulation is important in the permanent withdrawal of cardiomyocytes from the cell cycle. PMID- 9344765 TI - Repeated catecholamine surges alter cardiac isomyosin expression but not protein synthesis in the rat heart. AB - To assess the role of intermittent beta-adrenergic stimulation on alpha-myosin heavy chain expression and cellular hypertrophy, we studied the effect of intermittent dobutamine on myosin heavy chain isoform distribution and protein synthesis in the heterotopic rat heart preparation. This model allows the analysis of a pharmocologic stimulus in isolation from the mechanical load on the myocardium induced by the drug. Intermittent administration of dobutamine resulted in elevated alpha-MHC levels (75 +/- 12%) compared to control (55 +/- 10%; X +/- s.e.; P<0.05) transplanted hearts. This effect was not altered by alpha-receptor blockade with terazosin (72 +/- 8%). Intermittently pacing the transplanted hearts at the same rate as observed with dobutamine alone, also elevated alpha-MHC levels (70 +/- 5%). In contrast, total protein synthesis in the transplanted hearts was not altered with any of the drug or pacing interventions compared to control hearts. These data suggest that intermittent beta-receptor stimulation and/or intermittent increased heart rate contribute to altered patterns of myosin heavy chain expression. However, increases in cardiac mass and protein synthesis are probably mediated by hemodynamic factors rather than catecholamine stimulation. PMID- 9344767 TI - Competition between lactate and fatty acids as sources of ATP in the isolated working rat heart. AB - Fatty acid oxidation is generally considered the major source of energy in the heart, although lactate oxidation can be a major contributor to ATP production, depending on the concentration and availability of other competing substrates. In this study, isolated working rat hearts were used to directly determine the relationship between lactate and fatty acid oxidation to overall ATP production from exogenous sources. A range of lactate from 0.5 to 8.0 mM lactate was added to hearts perfused with buffer containing 5.5 mM glucose, and either 0.4 or 1.2 mM palmitate over a 100 min period. Rates of glycolysis, glucose oxidation, lactate oxidation, and palmitate oxidation were determined. In the presence of 0.5 mM lactate and 0.4 mM palmitate, lactate oxidation provided 17% of the ATP production and palmitate oxidation provided 68%, with the remainder coming from glucose oxidation and glycolysis. In the presence of 0.4 mM palmitate, an increase in lactate from 0.5 to 8.0 mM increased the steady state rates of lactate oxidation from 1239+/-236 to 5247+/-940 nmol/min/g dry weight, respectively. The contribution of lactate oxidation to total ATP production increased to 37%, with palmitate oxidation now contributing only 52% of the total ATP produced. At 8.0 mM lactate and 1.2 mM palmitate, lactate oxidation contributed 13% of the total ATP production, while palmitate oxidation contributed 81%. This data demonstrates that under near physiological conditions of lactate (0.5 mM) and fatty acids (0.4 mM), the preferred energy substrate of the heart remains to be fatty acids, and that only at high levels of lactate, such as can be observed during exercise or severe stress, does lactate oxidation become a significant source of ATP production. PMID- 9344768 TI - Effects of chronic beta-adrenergic receptor stimulation in mice. AB - The goal of the present study was to determine the effects of chronic beta adrenergic receptor stimulation with isoproterenol (ISO) on cardiac tissue, systemic trophic changes and on beta-adrenergic receptor desensitization in mice. Mice (n=36) received continuous ISO (30 microg/g/day) via osmotic minipump for 13 days. Left ventricle (LV)/body weight (BW) ratio was increased by 27% in ISO v control (CON) mice. The extent of cardiac hypertrophy induced by chronic ISO was offset in part by concomitant increases in body weight, which were greater in ISO than CON mice (22 v 8%), and occurred with increases in both muscle mass and brown fat to BW ratios. Histological analysis of mice revealed a three-fold increase in subendocardial interstitial connective tissue with no evidence of acute cellular necrosis or chronic inflammation. Acute i.v. ISO challenges induced dose-dependent increases in LV fractional shortening (FS) and ejection fraction (EF) using echocardiography (9 MHz), which were attenuated after chronic ISO, i.e. physiological desensitization was observed. Cellular mechanisms of beta adrenergic receptor desensitization included decreases in beta-adrenergic receptor density (-49%) and decreased basal (-45%) and ISO-stimulated (-61%) adenylyl cyclase activities. Lesser decreases in forskolin-stimulated adenylyl cyclase activity (-16%) and adenylyl cyclase mRNA levels for both type V (-17%) and type VI (-23%) isoforms were observed following chronic ISO. Thus, chronic ISO (30 microg/g/day) induced cardiac hypertrophy without cellular necrosis, increased weight gain and clear physiological desensitization in mice, with more extensive biochemical mechanisms than expected from simple catecholamine-specific (homologous) desensitization. PMID- 9344769 TI - Role of the sarcoplasmic reticulum in contraction and relaxation of immature rabbit ventricular myocytes. AB - Previous indirect studies of newborn hearts have suggested a diminished functional role of the SR and a greater dependency upon trans-sarcolemmal Ca2+ fluxes to directly elicit contraction and promote relaxation. We tested the hypothesis that the SR in newborn rabbit hearts is functionally incompetent by measuring contraction and relaxation in ventricular myocytes isolated from the hearts of 1-2-day-old (newborn), 10-12-day-old (juvenile) and >150-day-old (adult) rabbits. Electrically stimulated twitch characteristics were compared to those elicited by the rapid application of 10 mm caffeine in the presence and absence of functional sarcolemmal Na-Ca exchange (disabled using a Na+- and Ca2+ free extracellular solution). During steady state, electrically-induced contractions were lower in amplitude in newborn and juvenile compared to adult myocytes (2.9+/-0.5 and 3.4+/-0.3 v 8.5+/-0.9% of resting cell length, respectively; n=24-29) and relaxation was slower in immature myocytes (t0.75 values: newborn 250+/-20; juvenile 240+/-10; adult 130+/-20 ms, n=14-21). Contrary to our hypothesis, caffeine triggered sufficient SR Ca2+ release from immature myocytes to elicit contractions of similar magnitude to adults (newborn 12.8+/-1. 1; juvenile 14.0+/-0.9; adult 15.0+/-1.6% of resting cell length, n=25 29). The amplitude of indo-1 Ca2+ transients during steady-state twitch was 36+/ 12% of the maximal caffeine-induced Ca2+ transient in newborns (n=6) and 59+/-4% in adults (n=6). Caffeine slightly prolonged relaxation in adult myocytes (t0. 75=200+/-30 ms), but accelerated relaxation in newborn and juvenile myocytes (t0.75=180+/-20 and 150+/-30 ms, respectively). When both the SR and Na-Ca exchanger were disabled, the rate of relaxation (attributable to the sarcolemmal Ca2+-ATPase and mitochondrial Ca2+ uniporter) of newborn and juvenile myocytes was significantly faster than in the adults (1660+/-210 and 3030+/-180 v 4530+/ 310 ms, respectively; n=14-21). We conclude that neonatal and adult rabbit ventricular myocytes have comparable SR Ca2+ load, but neonatal cells exhibit smaller fractional SR Ca2+ release during steady-state contractions and greater Ca2+ removal by sarcolemmal Na-Ca exchange during relaxation. PMID- 9344770 TI - Cobra venom cardiotoxin induces perturbations of cytosolic calcium homeostasis and hypercontracture in adult rat ventricular myocytes. AB - The effects of Cobra venom cardiotoxin (CTX) on the cellular morphology, twitch amplitude and intracellular calcium ([Ca2+]i) of the ventricular myocytes were studied. [Ca2+]i and twitch amplitude were determined with a fluorometric ratio method using Fura-2/AM and Calcium Green-1 as calcium indicators, and a videomicroscopic technique, respectively. Addition of 0.001-1 microM CTX led to a time-dependent loss of rod shaped cells, beginning at 1 min, and remaining stable by 20 min. CTX 1 microM initially caused a transient augmentation in amplitude of the electrically induced-[Ca2+]i transient and twitch amplitude in the single cardiac myocyte. This was followed by a prolongation in duration of [Ca2+]i. Eventually, cells became inexcitable and abruptly underwent contracture, and [Ca2+]i remained elevated. In the absence of electrical stimulation, 1 microM CTX induced a Ca2+ spike followed by a sustained elevation of [Ca2+]i, an effect different from that of 40 mm KCl or 10 mm caffeine, which caused a transient elevation in [Ca2+]i. Digital imaging microscopy of Calcium Green-1 fluorescence revealed that the increase in [Ca2+]i was accompanied by changes in cell shape without leakage of fluorescence dye in the early stage after administration of the toxin. In the absence of [Ca2+]o, the initial [Ca2+]i spike was reduced, but the second phase of elevation of [Ca2+]i still occurred. In addition, experiments using Mn2+ quench technique suggested that Ca2+-influx was induced by CTX, and that both ryanodine and thapsigargin, known to deplete Ca2+ from its intracellular pool, abolished the second phase of the elevation of [Ca2+]i. The effects of cardiotoxin were abolished by 10 mM Ni2+ and 10 mM -Ca2+-o, but not by 5 microM verapamil. In conclusion, the observations indicate that CTX causes an initial increase followed by a second sustained elevation in [Ca2+]i, which is accompanied by changes in cell shape-from rod to round-and hypercontracture. The initial [Ca2+]i spikes were attributed to the extracellular Ca2+ influx, while the second [Ca2+]i elevation was related to internal Ca2+ release. The high [Ca2+]i may be responsible for hypercontracture and cell death. Further studies are needed to verify it. PMID- 9344771 TI - The effects of hypertrophy and diabetes on cardiac pyruvate dehydrogenase activity. AB - Alterations in substrate selection and utilisation are characteristics of heart failure of different etiologies and these changes may be involved in the development of contractile dysfunction. Regulation of pyruvate dehydrogenase (PDH) is crucial in determining the relative contribution of glucose oxidation to energy production; however, the role of PDH in the development of heart failure has not been clarified. In this study, we present a reliable and simple method for assaying both the active and total forms of PDH (PDHa and PDHt respectively) in cardiac tissue, and have compared the effects of pressure overload hypertrophy and diabetes on PDH activity. PDHa and PDHt were measured in extracts of hypertrophied hearts after 5 weeks of pressure overload or in hearts after 7 weeks following induction of diabetes. There was no significant change in PDHt in the hypertrophied group, but the fraction of PDH in the active form significantly decreased from 61+/-1% in controls to 36+/-1% (P<0.05). Following diabetes, there was a decrease in the ratio of PDHa:PDHt from 60+/-3% to 11+/-1% (P<0.0001) and PDHt activity -6.2+/-0.9 to 2.7+/-0.4 micromol/min/g wet weight (P<0.02)]. This study reports for the first time that (i) concomitant with the development of compensated hypertrophy, there is a decrease in the fraction of PDH in the active form; and (ii) in the diabetic heart, there is marked decrease in total PDH activity in addition to a decrease in the fraction of PDH in the active form. These results indicate that myocardial substrate delivery to the mitochondria may be impaired in both hypertrophy and diabetes, which may lead to the energy depleted state observed in heart failure. PMID- 9344772 TI - Low concentrations of hydrogen peroxide improve post-ischaemic metabolic and functional recovery in isolated perfused rat hearts. AB - The aim of the present study was to test the hypothesis that low concentrations of hydrogen peroxide (H2O2) have a beneficial effect on post-ischaemic myocardial recovery. Functional and metabolic measurements were performed in isolated buffer perfused rat hearts exposed to 30 min perfusion with 0 (control group A), 25, 50, 100 or 200 microM H2O2 or 30 min global ischaemia followed by 30 min reperfusion with 0 (control group B), 25, 50 or 100 microM H2O2. Catalase (200 U/ml) was added as scavenger during reperfusion with 25 microM H2O2. Non-ischaemic perfusion: All concentrations of H2O2 induced an immediate vasodilatation, which was maintained in the 50 microM group, but it was followed by vasoconstriction in the 100 and 200 microM group. Left ventricular developed pressure (LVDP) was significantly increased at the end of perfusion in the 50 microM group compared to the control group. Exposure to 100 and 200 microM H2O2 significantly decreased LVDP and increased end-diastolic pressure. ATP was reduced in the 100 microM group. Post-ischaemic perfusion: Exposure to 25 microM H2O2 caused improved coronary flow during the first 20 min of reperfusion compared to the control group (accumulated coronary flow; 235.5 +/- 10.8 v 172.7 +/- 8.6 ml). LVDP was significantly higher in the 25 microM group compared to the control (59.8 +/- 10.2 v 22.1 +/- 7.3 mmHg), and end-diastolic pressure was significantly lower (32.1 +/- 19.6 v 78.8 +/- 2.2 mmHg) at the end of reperfusion. Improved recovery was not observed in the group exposed to 25 microM H2O2 plus catalase. Treatment with 25 microM H2O2 caused significantly improved recovery of tissue ATP and creatine phosphate. In conclusion, the present study showed that exposure to 25 microM H2O2 improved post-ischaemic recovery in hearts subjected to global ischaemia. PMID- 9344774 TI - NADPH-oxidase-dependent superoxide production by myocyte-derived H9c2 cells: influence of ischemia, heat shock, cycloheximide and cytochalasin D. AB - Extracellular oxygen radicals produced by H9c2 rat heart cells in monolayer cultures during ischemia and subsequent reoxygenation were monitored using the luminol-horseradish peroxidase-enhanced chemiluminescence technique. As expected, the photon count diminishes during ischemia but again rapidly attains normal values following reoxygenation. In the presence of superoxide dismutase, this photon emission is repressed, as is also the case in the presence of diphenylene iodonium, a specific inhibitor of NADPH-oxidase activity. Thus, the conclusion seems justified that H9c2 rat heart cells in monolayer cultures produce superoxide radicals extracellularly due to an NADPH oxidase-like action. In order to characterize this extracellular superoxide-generating system, we determined its sensitivity to increased temperatures, inhibition of protein synthesis and perturbations of cytoskeletal structures. Heat shocks result in a delayed inactivation of the NADPH oxidase activity followed by recovery, the kinetics of which depend on the imposed heat shock temperature. This inactivation is independent of protein synthesis and actin cytoskeletal structures, but the recovery of the enzyme's activity is dependent on these entities. PMID- 9344773 TI - Proliferating cell nuclear antigen (PCNA), DNA synthesis and mitosis in myocytes following cardiac transplantation in man. AB - Cardiac transplantation is characterized by rejection, myocyte loss, interstitial and replacement fibrosis, and loading abnormalities. These modifications contribute to enhance mural and muscle cell stress, activating reactive growth processes in myocytes and interstitial cells. However, it is unknown whether cell cycle related gene product, such as PCNA, and DNA synthesis are stimulated under these conditions. Therefore, 62 endomyocardial biopsies obtained from 17 patients who underwent cardiac transplantation were examined for the immunocytochemical detection of PCNA protein in myocyte and non-myocyte nuclei. In addition, tissue samples were labeled in vitro with bromodeoxyuridine (BrdU) to document ongoing DNA synthesis. The presence of mitotic images in myocytes and non myocytes were also examined. Biopsies were collected from 1-768 days after surgery. Histologic examination of tissue sections documented that PCNA labeling involved nearly 30% of myocyte nuclei in all patients. Similar percentages of PCNA labeling were detected in interstitial cells, lymphocytes and mononuclear infiltrates. DNA synthesis in myocytes and connective tissue cells was observed in nine and 14 subjects, respectively. BrdU positive lymphocytes and mononuclear infiltrates in 13 cases. Three mitotic figures in myocyte nuclei were identified. PCNA, BrdU labeling and mitosis were not detected in eight myocardial samples obtained from control hearts. These results suggest that the evolution of the transplanted heart involves the expression of a gene which is implicated in DNA replication. The presence of ongoing DNA synthesis and mitosis support the notion that proliferation of myocytes and non muscle cells may be a component of ventricular remodeling after cardiac transplantation. PMID- 9344775 TI - Neonatal rat cardiomyocytes possess a large population of stable microtubules that is enriched in post-translationally modified subunits. AB - Microtubules (MTs) may be involved in several differentiation-specific cardiomyocyte functions, including myofibril organization, hypertrophic cell growth, and the negative regulation of heart cell contraction. The functional characteristics of neonatal rat heart cell MTs, which possess subsets that are enriched with either detyrosinated or acetylated tubulin subunits, were analysed. When compared with their non-myocyte neighbors, cardiomyocyte MTs were found to be more resistant to high concentrations (33 microM) of nocodazole (over 10-30 min), cold treatments (10 min to 8 h), or calcium (50-100 microM for 1-10 min, using detergent-extracted cells). These stable MTs were correlated with the modified MT population. Myocytes treated with 10 microM taxol for 4 h showed elevated levels of modified tubulin but only moderate MT bundling or rearrangement, while after an overnight treatment significant changes in intensity and distribution were noted. In contrast, non-myocytes on the same coverslip showed much greater MT rearrangements, but with only a moderate increase in the immunostaining intensity for modified MTs. Finally, hapten labeled tubulin that was microinjected into the myocytes showed incorporation rates that were similar to those of the non-myocytes in this study as well as to fibroblast rates determined in other studies, suggesting that the modified MTs in cardiomyocytes may be only moderately stable. Thus, a significant subset of MTs in neonatal cardiomyocytes are post-translationally modified, are resistant to depolymerization by a variety of agents, but are still turning over. These MTs may help define myocyte function during heart development. PMID- 9344776 TI - Beneficial effects of the 21-aminosteroid U 74389G on the ischemia-reperfusion damage in pig hearts. AB - 21-Aminosteroids (Lazaroids), acting as free radical scavengers and as membrane stabilizers, proved to have beneficial effects in various pathological conditions. In the present study we explored the effectiveness of one of these compounds, U 74389 G, in protecting pigs myocardium against the ischemia reperfusion damage induced by transient coronary occlusion achieved by clampling the left anterior descending coronary artery. Animals were divided into three groups: control, untreated and treated. Control animals were operated but not subjected to ischemia-reperfusion; the untreated group underwent to ischemia reperfusion without pharmacological treatment; while the treated group received the aminosteroid (4 mg/kg) before coronary occlusion and at the time of reperfusion. Specimens of myocardial tissue and blood samples were taken for morphological and biochemical studies. In the ischemic-reperfused myocardium of the untreated animals, the dominant morphological features were neutrophil infiltration, intercellular edema and severe swelling of mitochondria. All these alterations, notably neutrophil infiltration, were attenuated by aminosteroid treatment. As for the biochemical findings, the changes in adenine nucleotides and nucleosides levels, thus the reduction of energy charge, were reversed in the treated, but not in the untreated group. Myocardial concentration of malondialdehyde, which was undetectable in the control group, was raised in all the animals after reperfusion, but this effect was significantly less marked with aminosteroid treatment. In addition, the higher myocardial content of ascorbic acid and the reduced serum potential peroxidation exhibited by the treated animals compared with untreated group indicate an enhanced antioxidant protection induced by aminosteroid administration. On the other hand, the serum levels of myoglobin, cardiac troponin I and creatine kinase MB isoenzyme suggest the ability of the aminosteroid to attenuate the modifications of membrane permeability induced by ischemia-reperfusion injury. All these results lead to the conclusion that aminosteroid treatment, at least in the conditions of the present study, is effective in reducing the morphological and biochemical alterations occurring in ischemic-reperfused myocardium. PMID- 9344777 TI - Hypertrophy decreases cardiac KATP channel responsiveness to exogenous and locally generated (glycolytic) ATP. AB - This study tests the hypothesis that glycolytic regulation of KATP channel activity is altered in myocardial hypertrophy. Left ventricular (LV) subendocardial myocytes were isolated from cats with normal or left ventricular hypertrophied hearts (LVH). Saponin-permeabilized open cell-attached patch configurations of normal and LVH cells were exposed to an exogenous ATP consuming system containing hexokinase and 2-deoxyglucose. Phosphoenol pyruvate (PEP, substrate for the last ATP producing step in glycolysis) was applied extracellularly; ADP was present. In both cell types, KATP channels were activated in the absence of PEP, inhibited when PEP was added and activated again when PEP was removed, indicating the cells retained metabolic integrity and generated ATP in the proximity of their KATP channels. Single channel conductance in the absence of PEP was similar (70 pS, normal; 66 pS, LVH). However, LVH KATP channels showed enhanced activity (P0=0.50+/-0.03); normal (0.41+/-0.03) in PEP absence (P<0. 05). PEP responsiveness was reduced in LVH, with IC50, PEP increased to 23 microM; (11 microM normal). Lactate failed to activate KATP channels in both cell types. The concentration-P0 response curves obtained during exposure of open cells to exogenous ATP also revealed reduced responsiveness to ATP of LVH KATP channels (IC50, ATP=283 microM LVH; 93 microM normal). Our data indicate myocardial hypertrophy increases the maximal activity of KATP channels in the absence of ATP and reduces their responsiveness to ATP, including locally generated glycolytic ATP. These alterations in metabolic regulation of myocardial electrophysiology may contribute to diversity of action potential shortening in hypertrophied hearts during acute ischemia. PMID- 9344778 TI - Hemorrhage activates myocardial NFkappaB and increases TNF-alpha in the heart. AB - The heart is a tumor necrosis factor (TNFalpha) producing organ. Locally (v systemically)-produced TNFalpha likely contributes to myocardial dysfunction via direct suppression of myocardial contractile function, the induction of myocardial apoptosis, and the genesis of cardiac hypertrophy. Although recent studies have demonstrated increased myocardial TNFalpha following endotoxemia, it remains unknown whether shock, in the absence of sepsis, activates myocardial nuclear factor kappa B (NFkappaB, a TNFalpha transcription factor) and/or increases TNFalpha in the heart. To study this, rats were hemorrhaged and resuscitated, after which hearts were harvested and analysed for evidence of NFkappaB activation (electrophoretic mobility shift assay) and assayed for TNFalpha levels. Hemorrhage and resuscitation activated NFkappaB and resulted in a dramatic increase in myocardial TNFalpha. This study constitutes the initial demonstration that hemorrhagic shock activates the signaling mechanisms which culminate in increased myocardial TNFalpha. Indeed, this may have important clinical implications, since hemorrhage is a frequent complication of both iatrogenic and accidental trauma, as well as a potent instigator of multiple organ failure. PMID- 9344779 TI - Myocardial oxygen tension in isolated erythrocyte-perfused rat hearts and comparison with crystalloid media. AB - The isolated heart, typically perfused with crystalloid media equilibrated with >/=95% O2 to ensure adequate myocardial oxygen tension, is commonly used to study cardiac function. When hemoglobin is available for oxygen transport, equilibration with 21% O2 is considered adequate to meet metabolic demands. This study presents the measurement of myocardial pO2 in isolated hearts perfused with an erythrocyte suspension. Baseline myocardial pO2 in erythrocyte-perfused hearts was 16.4+/-3.5 mmHg (mean+/-s.e.). When compared to previous measurements of myocardial pO2 in isolated hearts perfused with crystalloid media, the use of erythrocyte suspensions resulted in a 10-fold lower level of myocardial pO2, while avoiding very low and high values. The standard use of 95% oxygen with crystalloid results in myocardial levels of oxygen far above those usually found in the presence of hemoglobin and room air. PMID- 9344780 TI - The novel heat-stable enterotoxin subtype gene (ystB) of Yersinia enterocolitica: nucleotide sequence and distribution of the yst genes. AB - The gene (ystB) encoding the novel subtype of the heat-stable enterotoxin (Y-STb) was cloned from the chromosome of a clinical isolate of Yersinia enterocolitica 84-50 (serotype O:5, biotype 1A) and the nucleotide sequence was determined. The ystB contained 216 base pairs that encoded a protein of 71 amino acid residues. The C-terminal 30 residues of the precursor protein exactly corresponded to the amino acid sequence of the Y-STb toxin, purified from the culture supernatant of the wild strain. Homology search revealed that there are 76.9% nucleotide sequence similarity between ystB and the Yersinia kristensenii ST gene, and 73.5% with the Y. enterocolitica prototype sequence of yst (ystA). When tested with the PCR generated ystB specific probe, 36 of 304 Y. enterocolitica strains from 18 countries hybridized with the probe. All the ystB probe positive strains belonged to biotype 1A and mostly to the so-called non-pathogenic serotype O:5, O:6, O:7,8 O:7,13 and O:10, while ystA was predominantly found among the pathogenic serotypes (78.5%). Out of 36 ystB gene positive strains, 18 were clinical origin from six countries, which were also positive in the suckling mice assay suggesting that ystB may play an important role in the pathogenesis, and the so called non-pathogenic serotypes could be virulent for human. PMID- 9344782 TI - Studies on the role of plasminogen activator in systemic infection by virulent Yersinia pestis strain C092. AB - Plasminogen activator is an outer membrane protease of Yersinia pestis encoded by the pla gene on plasmid pPst. Pla of the KIM-10 strain of Y. pestis appears to be required for the virulence from a subcutaneous (sc) but not an intraperitoneal (ip) or intravenous (iv) route of infection in mice. However, other strains of Y. pestis are highly virulent by the sc route yet lack pPst and pla. In this study, the pPst- Pestoides F strain was lethal to mice inoculated sc, with an LD50 (3 cfu), equal to that of C092, a virulent pPst+ strain. To analyse further the role of Pla in invasive infection, isogenic derivatives of C092, including one harboring pla with a frameshift mutation and another cured of pPst, were made. Although the ip LD50 of pPst- C092 and of the pla mutant were nearly identical to that of the wild type, the subcutaneous LD50 of the cured and mutant strains were 4 to 6 logs greater than that of wild type. Thus, pPst appears to be required for development of a lethal infection by some strains after sc inoculation but not after direct ip inoculation. Pla-associated virulence did not appear to be mediated by interference with the phagocyte chemoattractant C5a, as shown by the lack of correlation of C5a production with susceptibility to Y. pestis in C5a+ and C5a- congenic mice. In a footpad model of the early host response to subcutaneous infection, pPst- C092 proliferated at the subcutaneous injection site to a similar extent as did the wild type parent strain, and elicited a similarly large, local inflammatory response. However, the wild type was present at higher concentrations at more distant sites such as the popliteal lymph node and spleen. PMID- 9344781 TI - The hemolytic and complement-activating properties of pneumolysin do not contribute individually to virulence in a pneumococcal bacteremia model. AB - The virulence of pneumococcal capsular type 2 strain D39 and derivatives with mutations in the pneumolysin gene were examined in a mouse bacteremia model. In CBA/N-XID mice D39 is known to exhibit exponential growth in the blood until the death of the mice at 24 to 36 h. In contrast, PLN, a pneumolysin-deficient derivative of D39, reaches a plateau in growth that is maintained for several days. The growth patterns of D39 and PLN observed in CBA/N-XID mice were also observed in C3H/HeJ and C3H/HeOuJ mice, but not in 129/SvJ and C57BL/6J mice. These results demonstrate that the effect of pneumolysin on bacteremia is dependent on the genetic background of the mice. D39 derivatives with point mutations which abolish the cytotoxic or complement-activating properties of pneumolysin did not have major individual effects on virulence in CBA/N- XID and C3H/HeOuJ mice. A derivative with mutations affecting both the cytotoxic and complement- activating properties resulted in a modest, yet statistically significant, increase in survival time of i.v. challenged CBA/N-XID mice. However, the effect was less marked than that seen with PLN. These findings suggest that the virulence effects of pneumolysin in bacteremia must be due in part to properties other than hemolysis and complement fixation. PMID- 9344783 TI - Cell-mediated HTLV-I infection of a cervix-derived epithelial cell line. AB - There is considerable evidence that sexual transmission of human T-cell leukemia virus-I (HTLV-I) is mediated by virus-infected lymphocytes in genital tract secretions. However, it is not clear whether infection occurs through lesions in the genital tract epithelium or takes place via an intact epithelium. We have carried out experiments to test the hypothesis that sexual transmission of HTLV-I is initiated by lymphocyte-mediated infection of intact genital tract epithelia. To examine this question we added either free virus or HTLV-I producing MT-2 cells to cultures of a cervix-derived epithelial cell line, MS751. Although free virus did not infect MS751 cells, MS751 cells which had been coincubated with MT 2 cells became infected. These cultures produced about 50 pg/ml of HTLV-I p24 antigen per 10(6) cells over a 24 h period on the sixth day following exposure to donor T-cells. Proviral DNA could be detected in target MS751 epithelial cells by PCR. Infection of epithelia could be blocked, in a dose-dependent manner, by the sulfated polysaccharides dextran sulfate, heparin, and fucoidan, and by the enzymes fucosidase and mannosidase, but not by a number of other agents that were tested. Since MT-2 cells were observed to attach to the epithelial monolayer, we examined the ability of agents to inhibit adhesion. Adherence was inhibited by the same agents that inhibited infection. Based on these findings, we hypothesize that sexual transmission of HTLV-I may involve lymphocyte-mediated infection of genital tract epithelia and that lymphocyte adhesion to the epithelium is a critical event in transmission of HTLV-I. We speculate that a sugar moiety on the epithelium, possibly mannose or fucose, may be involved in adhesion of T-cells to epithelial cells. As sulfated polysaccharides block both adhesion and productive infection of the epithelium, these compounds might be used as active ingredients in a vaginal formulation to help prevent HTLV-I transmission. PMID- 9344784 TI - Some properties of nicked Vibrio vulnificus hemolysin. AB - Vibrio vulnificus, an opportunistic human pathogen, secretes the 50 kDa single chain hemolysin. When incubated with an exocellular protease from this vibrio, the 50 kDa hemolysin was cleaved in some peptides joined with the disulfide bond(s); the 40 kDa fragment and the small fragment(s) undetectable in SDS-PAGE. The nicked hemolysin induced comparable hemolysis through the same process as that of the intact toxin. However, the nicked hemolysin was found to be more stable against inactivation due to autoaggregation, so that it formed a larger precipitation zone in the single radial immunodiffusion test using the antiserum against the intact hemolysin. These results suggest that V. vulnificus hemolysin is modified to be a more hydrophilic protein by nicking, while it is not accompanied by loss of the hemolytic activity. PMID- 9344785 TI - Aeromonas spp. possess at least two distinct type IV pilus families. AB - Type IV pili have been purified from strains of most of the Aeromonas species associated with gastroenteritis (A. veronii biovar sobria, A. hydrophila, A. trota and A. caviae). They appear to be a related family (molecular mass of pilin 19 to 23 kDa) with a tendency to bundle-formation. Hence, we have designated them 'bundle-forming pili' (Bfp). A type IV pilus biogenesis gene cluster (tapABCD) recently cloned from a strain of A. hydrophila, however, encoded a 17 kDa pilin which differed significantly in its N-terminal amino acid sequence from the Bfp pilins. This paper describes the cloning of part (tapA and approximately 20% of tapB) of a homologous pilin gene cluster from a Bfp-positive strain of A. veronii biovar sobria, and presents evidence that the entire pilin gene cluster (tapABCD) is present in this strain. The predicted N-terminal amino acid sequence of the pilin encoded by the A. veronii biovar sobria tapA differed markedly from the corresponding sequence of its Bfp pilin, and those of the Bfp purified from other Aeromonas strains and species. Probing with tapA and tapD genes showed that these Bfp-positive Aeromonas strains also possessed the Tap gene cluster. TapA proteins of A. veronii biovar sobria and A. hydrophila shared 53% identity and 63% homology. We conclude that Aeromonas species are potentially able to express at least two distinct families of type IV pili (Bfp and Tap). PMID- 9344786 TI - Pseudomonas aeruginosa entry into Caco-2 cells is enhanced in repairing wounded monolayers. AB - Human respiratory cells participating in the repair of epithelial wounds have been shown to be highly susceptible to Pseudomonas aeruginosa adherence. To ascertain whether such susceptibility is a common feature of different repairing epithelial cells, Caco-2 cell monolayers were chemically injured, reincubated for 48 h to partially repair and exposed to bacteria. Cells edging the wounds that spread and migrate to re-establish cell confluence were called 'repairing cells' while cells far from the wounds were called 'non-repairing cells'. By light microscopy, bacteria were seen to adhere to and to enter into both repairing and non-repairing cells. The percentage of intracellular bacteria in repairing cells was significantly higher than in non-repairing cells. The higher susceptibility of repairing monolayers to bacterial entry was confirmed by the gentamicin exclusion assay. P. aeruginosa entry into Caco-2 cells was greatly enhanced in non-injured confluent monolayers treated with EDTA to disrupt intercellular junctions. As tight junction disfunctions have been described during the wound repair process, we speculate that exposure of basolateral receptors to bacterial ligands may account for the enhancement of P. aeruginosa internalization by repairing monolayers. PMID- 9344787 TI - Electronic Spectroscopy of UO AB - Spectra for gas-phase uranium oxide have been analyzed to obtain molecular constants and provide a partial atlas of the visible and near infrared bands for the isotopomers 238U16O, 238U18O, 235U16O, and 235U18O. Nineteen bands of the 238U16O, 238U18O, and 235U18O isotopomers were rotationally analyzed. With few exceptions, Omega assignments were unambiguously determined from observations of the first lines in at least two rotational branches. Accurate term values and rotational constants are reported. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344788 TI - New Band Systems of the YbCl Molecule AB - The thermal emission spectrum of the YbCl molecule, lying in the 3800-6000 A region, has been photographed for the first time at a reciprocal linear dispersion of 7.3 A/mm. More than 196 violet degraded bands have been recorded on a 2-m plane grating spectrograph in first order and assigned to the vibrational schemes of six systems, viz. A, B1, B2, C1, C2, and D. While the systems A, C1, C2, and D consist of single headed bands, the systems B1 and B2 consist of double headed bands. The vibrational analysis performed suggests that these systems arise from the ground state (2Sigma+) of YbCl molecule. The systems C1-X, C2-X, and D-X have been analyzed for the first time. More precise vibrational constants for the relevant states have been determined. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344789 TI - High-Resolution Infrared Study of the Fundamental Bands of Deuteroiodoacetylene AB - The high-resolution infrared spectrum of deuterated iodoacetylene has been measured in the region 200-2700 cm-1. In addition to all five fundamentals, the bands 2nu40 and 2nu50 have been identified. The measurements were carried out by using a Fourier transform spectrometer at room temperature with instrumental resolution of about 0.0020 cm-1. The ground state rotational constants B0 = 0.0970742961(72) cm-1, D0 = 1.38482(20) x 10(-8) cm-1, and H0 = -1.47(10) x 10( 15) cm-1 have been determined by combining 462 ground state combination differences from the same bands mentioned above with accurate MW data from the literature. In addition, the molecular constants for all the fundamental levels have been obtained. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344790 TI - Fourier Transform Microwave Spectroscopy of 13C-Substituted CCS Radicals AB - The rotational spectral lines of the 13C isotopic species of CCS (13CCS, C13CS, and 13C13CS) have been observed using a Fourier transform microwave spectrometer in combination with a pulsed-discharge nozzle. The hyperfine-resolved JN = 10-01, JN = 21-10, and JN = 32-21 transitions have been observed in the 11-, 22-, and 33 GHz regions, respectively, with an accuracy of about 5 kHz. The observed transition frequencies for 13CCS and C13CS are analyzed simultaneously with millimeter-wave data, and the hyperfine interaction constants for both species are determined accurately. Astronomical implications for these radicals are discussed. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344791 TI - Absolute Line Intensities in the 2nu3 Band of 16O12C32S AB - The strengths of 100 lines in the 2nu3 band of 16O12C32S have been measured at high resolution in the spectral range 4069-4118 cm-1, using a tunable difference frequency laser spectrometer. These intensities were obtained by fitting Voigt profiles to the measured shapes of the lines. The vibrational transition moment [(2.141 ± 0.020) x 10(-2) D] and the absolute intensity (16.19 ± 0.24 cm-2 atm-1 at 296 K) of the 2nu3 band of 16O12C32S are determined from these linestrength measurements. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344792 TI - Interaction of Vibrational Fundamental and Combination States of Ethylene in the 3 &mgr;m Region AB - High-resolution IR spectra of ethylene cooled in a molecular jet have been investigated revealing perturbations of some rovibrational lines of nu9 and nu11 fundamentals due to resonant Coriolis interactions. In the range of J up to 4, two combination states, nu3 + nu8 + nu10 and nu6 + nu8 + nu10, were shown to be the strongest local perturbers. Borrowing of dipole moments due to third-order Coriolis interaction was observed and the appropriate rovibrational lines of the combination bands were assigned. Modeling of the observed spectra was achieved using the power expansion of the Hamiltonian and the dipole moment operator in the frame of tensorial formalism of the symmetry point group of the molecule. The coupling of 32 vibrational combination states with the nu9 and nu11 fundamentals via cubic anharmonicity and Coriolis interaction was considered. Many new coupling parameters have been determined. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344793 TI - Dispersed Fluorescence Spectroscopy of the ZnC2H5 Free Radical AB - Dispersed fluorescence spectra of the zinc monoethyl radical are presented. These have allowed vibrational frequencies of several modes in the ground electronic state of this molecule to be determined for the first time. In particular, substantial activity in both the Zn-C stretch and the Zn-C-C bend has been observed, with the frequencies for these modes being deduced as 387 and 180 cm-1, respectively. With the aid of the dispersed fluorescence data it has also proved possible to make reasonable assignments for several bands in the excitation spectrum. ZnC2H5 was prepared in these experiments by a dc electrical discharge through an argon/diethyl zinc mixture in a supersonic nozzle. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344794 TI - First Spectroscopic Observation of the CdC2H5 Radical AB - We report the first spectroscopic observation of the monoethyl cadmium free radical, CdC2H5, using laser-induced fluorescence spectroscopy. This molecule was prepared in a supersonic jet by reaction of cadmium atoms with an ethyl precursor in a recently developed dual-channel electrical discharge nozzle. The laser excitation spectrum of CdC2H5 consists of a single, unresolved, band attributed to the A-X origin transition. However, the dispersed fluorescence spectrum obtained by laser-pumping the origin transition does show some vibrational structure. A short progression in the Cd-C stretching mode has been observed giving a frequency of 280 cm-1 for this mode. Comparison with the related molecule ZnC2H5 shows that the increase in reduced mass is not sufficient to account for the much lower metal-carbon stretching frequency in CdC2H5 and the evidence therefore points to a substantially weaker metal-carbon bond in the latter. The absence of vibrational structure in the excitation spectrum of CdC2H5 is attributed to the depopulation of excited vibronic levels by predissociation. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344795 TI - Measurements of H216O Linestrengths and Air-Induced Broadenings and Shifts in the 815-nm Spectral Region AB - The linestrengths for 40 absorption lines of H216O water vapor that were located between 813 and 820 nm were measured; most of these lines were selected for their potential usefulness in laser remote measurements of atmospheric humidity using the differential absorption lidar technique. The air-induced pressure-broadening coefficients were also measured for 32 of these lines and the air-induced pressure shift coefficients were measured for 29 lines. These spectroscopic parameters were derived from spectra obtained with an AlGaAs diode laser and two long-path absorption cells. Collisional narrowing effects were observed and were accurately described by a Galatry profile. Comparisons were made with previous experimental work or theoretical calculations as available. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344796 TI - The Rotational Spectrum of CBrClF2 (Halon BCF): II. The Lowest Excited Vibrational States and Nuclear Quadrupole Coupling Tensors AB - The analysis of the millimeter-wave rotational spectrum of the well-known halon molecule CBrClF2 was extended to rotational transitions in the excited vibrational states v9 = 1 (307 cm-1) and v5 = 1 (337 cm-1). The two states are appreciably coupled by a- and b-axis Coriolis interaction which was treated explicitly and spectroscopic constants were determined from measurements at frequencies from 121 to 330 GHz and J" from 18 to 111. The present work, together with our previous study (1996. J. Mol. Spectrosc. 177, 240-250), results in accurate constants in the rotational Hamiltonian for the four lowest vibrational states of C79Br35ClF2 and C81Br35ClF2. The double nucleus hyperfine structure in low-J transitions of the two 35Cl isotopomers of CBrClF2 was also measured in the region 3-14 GHz with a supersonic beam, cavity Fourier transform microwave spectrometer. All constants in both the inertial and the principal nuclear quadrupole coupling tensors have been determined for both Br and Cl nuclei. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344797 TI - High-Resolution Study of the Infrared Spectrum of H3SiI in the Regions 330-680 and 1070-1360 cm-1: Accurate Determination of the Ground State Constants AB - High-resolution FTIR spectra of H3SiI have been recorded in the regions 330-680 cm-1 (nu3/nu6) and 1070-1360 cm-1 (2nu6). A detailed rovibrational study was carried out for the nu3 and nu6 fundamental bands, 2nu6-/+2 and 2nu60 overtone bands, and two hot bands, 2nu6+/-2 - nu6+/-1 and 2nu60 - nu6. A local resonance between the v6 = 2 and v2 = v3 = 1 states was observed. Ground state combination differences deduced from the nu3, nu6, 2nu6-/+2, and 2nu60 bands allowed us to obtain accurate B0, DJ0, and DJK0 constants. The nu6, 2nu6+/-2 - nu6+/-1, and 2nu6-/+2 bands were used for the experimental determination of the A0 and DK0 constants, whereas the hot band 2nu60 - nu6 served to make the internal calibration coherent. Ground state differences DeltaK(J) = E0(J, K) - E0(J, K - 3) were calculated with K up to 12. By a least-squares fit, we obtained the following results:A0 = 2.8426037(14) cm-1 and DK0 = 2.75840(99) x 10(-5) cm 1.Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344798 TI - Study of the v3 = 1 State of 80SeF6 by Fourier Transform Spectroscopy AB - The Fourier transform infrared spectrum of monoisotopic 80SeF6 has been recorded in the 760-792 cm-1 region with an effective resolution of ca. 2.3 x 10(-3) cm-1. The 80SeF6 sample was prepared by burning monoisotopic 80Se powder (99.2%) in an excess of fluorine. The analysis of infrared transitions of the nu3 band enabled the determination of parameters of the Hamiltonian developed up to the third order and the fourth order. The standard deviation obtained is equal to 4 x 10( 4) cm-1 for the third-order development and 3.2 x 10(-4) cm-1 for the fourth order development. In the two analyses, 2900 lines were assigned and fitted. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344799 TI - The Microwave Spectrum of the Methanol Dimer for K = 0 and 1 States AB - The rotational spectrum of (CH3OH)2 has been observed in the 8 to 24 GHz region with a pulsed-beam Fabry-Perot cavity Fourier-transform microwave spectrometer. Previously we demonstrated that each transition of the a-type R(J), Ka = 0 is split into 15 states of the 16 theoretically expected states by tunneling motions. Here we show that the K = 1 states are split into the 16 expected states through the assignment of the Ka = 1 a-type transitions and DeltaKa = 1 b-type transitions. The internal-rotation analysis of the two inequivalent methyl groups presented here was guided by the previous experimental observations and theory for multidimensional tunneling, which predicts 16 tunneling components for each R(J) transition from 25 distinct tunneling motions. The effective barrier to internal rotation for the donor methyl group of (CH3OH)2 is V3 = 183.0 cm-1, and is one-half of the value for the methanol monomer (370 cm-1), while the barrier to internal rotation of the acceptor methyl group is 120 cm-1, one-third of the methanol monomer. The structure of the methanol dimer complex is similar to that of water dimer with a hydrogen bond distance of 1.96(2) A and tilt of the acceptor methanol of 77(2)degrees from the O-H-O axis (one standard deviation uncertainty). This structure shows good agreement with the angular orientation of the methyl groups derived in the internal-rotation analysis. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344800 TI - Exchange Energy on Alkali Diatomic Molecules: Period Effects AB - The exchange energy in the long-range near-dissociation region is analyzed for the ground electronic states of homo- and heteronuclear alkali diatomic molecules. The analysis is based on the use of the functional form Ae-ar to represent this exchange term. A period effect is observed in the values of both parameters and the exponent parameter a appears to approach an asymptotic limit for the heavier molecules. The results obtained in this study also suggest that some care must be taken in using even this function to represent the exchange energy. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344801 TI - Equilibrium Structure and Spectroscopic Constants of Dichloroethyne: An ab Initio Study AB - High-level ab initio calculations with large basis sets are reported for dichloroethyne, ClCCCl. Based on CCSD(T)/cc-pVQZ results, an empirically corrected theoretical equilibrium geometry is derived: re(CC) = 1.2025(6) A and re(CCl) = 1.635(5) A. Correlated harmonic (CCSD(T)/cc-pVQZ) and anharmonic (MP2/TZ2Pf) force fields provide theoretical predictions for the fundamental vibrational wavenumbers and many other spectroscopic constants for the four most important isotopomers of dichloroethyne. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344802 TI - Analysis of the Infrared Spectra of the Fundamentals nu3 and nu6 of 12CD3I and 13CD3I AB - The high-resolution infrared spectra of the lowest fundamental bands nu3 and nu6 of 12CD3I and 13CD3I have been measured using a Fourier transform spectrometer. The bands are analyzed on one hand by taking into account the Coriolis resonance nu3/nu6 and on the other hand without this Coriolis effect. The molecular constants obtained for the two vibration modes are introduced and a discussion of the influence of the Coriolis interaction to the parameter set is shortly outlined. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344803 TI - High-Resolution Coherent Raman Spectra of Vibrationally Excited 14N2 and 15N2 AB - At an effective resolution of 0.001 cm-1, we measured coherent anti-Stokes Raman spectra of electrically discharged 14N2 and 15N2 in the electronic ground state X1Sigmag+, specifically Q branches of bands with Deltav = 1 up to v' = 8 for 14N2 and v' = 7 for 15N2, and O and S branches of the fundamental band of 15N2. Account is taken of small wavenumber shifts due to pressure, AC Stark, and |chi|2 interference effects. Separate fits of the Q-branch data of each isotopic variant, combined with selected data from the literature, yield term coefficients Ykl and Ukl or potential-energy coefficients cj that reproduce wavenumbers of measured spectral lines generally within 0.004 cm-1. The value of the harmonic vibrational parameter omegae is 2358.5402(4) cm-1 for 14N2 and 2278.7913(7) cm-1 for 15N2. Efforts to combine spectral data of both isotopic variants to distinguish adiabatic and nonadiabatic effects arising from incomplete separation of electronic and nuclear motion in N2 were unsuccessful. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344804 TI - High-J Rotational Transitions of NNO Measured with the NAIR Terahertz Spectrometer AB - A BWO-based terahertz spectrometer has been constructed and tested by measuring some high-J rotational transitions of NNO in the frequency region from 620 to 730 GHz. The observed line center frequencies confirm the previously reported ones obtained by FTIR spectroscopy. The high sensitivity of the present spectrometer allowed us to record spectra of singly substituted isotopomers, 15NNO, N15NO, NN18O, and NN17O, in natural abundance. The rotational and centrifugal distortion constants have been revised. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344805 TI - Microwave Spectrum, Nuclear Quadrupole Coupling Constants, and Structure of Bromodifluoromethane AB - The microwave spectrum of bromodifluoromethane, CHBrF2 (Halon 1201) has been studied for the first time from 7 to 40 GHz. A least-squares analysis of the observed c-type transition frequencies gave rotational and centrifugal distortion constants and components of the bromine nuclear quadrupole coupling constant tensor in the principal axes system as follows: A = 10199.7186(62) MHz, B = 2903.4150(26) MHz, C = 2360.1521(23) MHz, DeltaJ = 0.660(14) kHz, DeltaJK = 2.87(11) kHz, DeltaK = 8.95 kHz, deltaJ = 0.1344(24) kHz, deltaK = 3.22(15) kHz, chiaa = 521.281(92) MHz, chibb - chicc = -38.32(9) MHz, and |chiac| = 187.1(26) MHz for the 79Br species; A = 10199.5567(54) MHz, B = 2876.5588(20) MHz, C = 2342.3796(18) MHz, DeltaJ = 0.652(12) kHz, DeltaJK = 2.77(9) kHz, DeltaK = 8.21(61) kHz, deltaJ = 0.1300(19) kHz, deltaK = 2.97(13) kHz, chiaa = 435.61(10) MHz, chibb - chicc = -32.08(8) MHz, and |chiac| = 148.5(29) MHz for the 81Br species. The structural parameters are calculated from all these rotational constants and the electronic properties of the carbon-bromine bond in bromodifluoromethane are evaluated from the observed nuclear quadrupole coupling constants. These molecular properties are compared with those of other related molecules. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344806 TI - Millimeter-Wave Spectrum of the NO Dimer AB - The pure rotational spectrum of the NO dimer, (14N16O)2, has been studied in the 225-400 GHz millimeter-wave region by means of direct absorption in a supersonic jet expansion. Previously, only 4 rotational transitions of (NO)2 had been measured in the microwave region. To these, we have added 66 new millimeter-wave transitions with values of J ranging from 4 to 16 and of Ka from 2 to 7. Most transitions were observed as single lines, but a few showed partially resolved hyperfine structure. The analysis of these data, in combination with the previous microwave transitions and an extensive set of infrared (nu1 band) measurements, has resulted in a greatly improved set of parameters for the ground state of the NO dimer, including the first sextic distortion parameters. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344807 TI - Sub-Doppler Infrared Spectra and Torsion-Rotation Energy Manifold of Methanol in the CH-Stretch Fundamental Region AB - The infrared spectrum of jet-cooled methanol in the CH-stretch fundamental region has been investigated by two sub-Doppler laser techniques: optothermally detected molecular-beam electric resonance and direct-absorption slit-jet spectroscopy. With the aid of microwave-infrared double resonance and ground state combination differences, 27 subbands in the frequency range 2967 to 3027 cm-1 have been assigned to the nu2 fundamental. Perturbation systems in the K' = 0 E, -1 E, and 2 E symmetry subbands have been analyzed to yield matrix elements of 0.013, 0.041, and 0.75 cm-1, respectively. The A-E torsional tunneling splitting for J = 0 of the nu2 vibration of -3.26 cm-1 is of opposite sign and a factor of three smaller in magnitude than the ground state value of +9.12 cm-1. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344808 TI - Study of the 41Piu, 51Piu, 61Piu, and 61Sigmau+ States of K2 by Polarization Labeling Spectroscopy AB - The polarization labeling spectroscopy technique is used to study excited singlet ungerade states of K2 in the energy range 26 400-30 200 cm-1 above the bottom of the ground state potential. Three states, 4-6(1)Piu, are characterized by sets of Dunham coefficients. The 6(1)Sigmau+ state with an unusual shape of the potential curve is also partially analyzed. Comparison with theoretical calculations is done. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344809 TI - Fourier-Transform Microwave Spectroscopy of the Argon-Diacetylene van der Waals Complex AB - The rotational spectrum of the argon-diacetylene van der Waals complex, produced in a supersonic molecular beam at 1 K, has been observed with a Fourier-transform microwave spectrometer. We observed 22 a-type rotational transitions with Ka up to 3. Three rotational constants, five centrifugal distortion constants, and one higher-order centrifugal distortion constant were determined precisely by least squares analysis. The complex is shown to have a planer T-shaped structure with C2v symmetry. The structural analysis provides that the Ar atom is located 3.68 A from the center of mass of diacetylene. Force constants for the van der Waals vibrations were determined from the centrifugal distortion constants. It has been found that this complex has a much steeper and more harmonic intermolecular potential than the argon-acetylene complex. Copyright 1997 Academic Press. Copyright 1997Academic Press PMID- 9344810 TI - The NCN 211310 Hot Band Measured by Infrared Laser Magnetic Resonance Spectroscopy PMID- 9344812 TI - Millimeter-Wave Spectra of ArDCN Observed with the Pulsed-Jet Expansion Technique PMID- 9344811 TI - Ar and Self-Pressure Broadening Coefficient of the R(11), 5;gn3 Line of 12C2H2 PMID- 9344813 TI - Accurate Equilibrium Structure and Electric Dipole Moment of HC9N: Predictions on the Basis of Large-Scale Coupled Cluster Calculations PMID- 9344814 TI - Spectroscopic Constants of Degenerate Vibrations of a C3upsilon Molecule Eligible for ab Initio Calculations of a Force Field PMID- 9344815 TI - A Determination of the Vibrational Anharmonicity of PD in Its X3Sigma;ms State PMID- 9344817 TI - Rotational Structure of the IR Spectra of Single SF6 Molecules in Liquid 4He and 3He Droplets PMID- 9344818 TI - High-Resolution Measurement of the nu2 Band of H15NO3 PMID- 9344816 TI - A Dispersed Fluorescence Investigation of the Low Frequency Vibrations of MgCCH(&Xtilde;2Sigma;pl) PMID- 9344819 TI - Cortical sites of sustained and divided attention in normal elderly humans. AB - Human brain mechanisms subserving attention have been assigned to prefrontal, midfrontal, and posterior parietal cortices, as well as to the anterior cingulate and the thalamus. To map these mechanisms in the brain, most studies have used selective attention tasks; few studies have mapped the brain under sustained or divided attention. The present study was designed to create maps of regional activity associated with sustained and divided attention using two different sensory modalities: visual checkerboard stimulation and vibrotactile stimulation of the right hand. Five cerebral PE-tomograms of 15O-labeled water uptake were acquired from 16 elderly healthy subjects during sustained or divided attention to the frequency of stimulation. To locate active brain regions, the t-statistic map of relative changes in cerebral blood flow was coregistered to the subjects' averaged brain MR images and to the standard Talairach brain coordinate system. Attention was associated with activity in two sites, the right middle frontal gyrus (Brodmann area 46) and the right inferior parietal lobule (Brodmann area 40). The frontal site was more active when the subjects attended to the visual stimulus and when the attention was divided, while the parietal site was more active during attention to the vibrotactile stimulus and during simple sustained attention. Our observations are consistent with the hypotheses (1) that the right posterior parietal attention center subserves attention to several sensory modalities and (2) that a cortical network of specific neuronal sites subserves both sustained and divided attention. These hypotheses must be tested in further studies. PMID- 9344820 TI - Anatomic localization and quantitative analysis of gradient refocused echo-planar fMRI susceptibility artifacts. AB - Functional magnetic resonance imaging (fMRI) techniques, such as echo-planar imaging, can permit rapid, sensitive, whole-brain measurements of local blood flow-induced MR signal changes seen during cognitive paradigms. Changes in blood oxygenation due to mismatch of flow and oxygen metabolism cause dynamic variations in microscopic susceptibility effects, leading to the blood oxygenation level-dependent (BOLD) signal measured by fMRI techniques. A related static macroscopic susceptibility effect is known to cause artifacts that attenuate the MR signal, leading to "blind spots" in some regions of brain adjacent to bone and air sinuses. The anatomical location, spatial extent, and magnitude of signal loss artifact are quantitated for a common whole-brain fMRI technique. Resting gradient-echo EPI studies were obtained in four healthy volunteers. Signal loss was primarily localized to inferior frontal regions (medial orbital gyri and gyrus rectus) and to inferior lateral temporal lobe (including part of fusiform gyrus) bilaterally. Increased echo time (TE) uniformly produced larger artifacts. The orientation of acquired slices and choice of phase-encoding direction influenced the location, shape, and extent of the artifacts. Regions of the brain with severe artifact may have attenuated activation signal, with potential implications for the design and interpretation of fMRI studies targeting activations in these areas. PMID- 9344821 TI - Fast and localized event-related optical signals (EROS) in the human occipital cortex: comparisons with the visual evoked potential and fMRI. AB - Localized evoked activity of the human cortex produces fast changes in optical properties that can be detected noninvasively (event-related optical signal, or EROS). In the present study a fast EROS response (latency approximately 100 ms) elicited in the occipital cortex by visual stimuli showed spatial congruence with fMRI signals and temporal correspondence with VEPs, thus combining subcentimeter spatial localization with subsecond temporal resolution. fMRI signals were recorded from striate and extrastriate cortex. Both areas showed EROS peaks, but at different latencies after stimulation (100 and 200-300 ms, respectively). These results suggest that EROS manifests localized neuronal activity associated with information processing. The temporal resolution and spatial localization of this signal make it a promising tool for studying the time course of activity in localized brain areas and for bridging the gap between electrical and hemodynamic imaging methods. PMID- 9344822 TI - FMRI studies of the supplementary motor area and the premotor cortex. AB - Brain activation patterns associated with three motor tasks, differing in the mode of movement selection, were studied in seven right-handed subjects, using functional magnetic resonance imaging (fMRI). The tasks consisted of sequences of finger movements in which the next finger was selected (i) according to a fixed sequence (FIX), (ii) in response to an external sensory cue (RAND), or (iii) on the basis of free, internal selection (SELF). Periods of hand relaxation (REST) alternating with the tasks served as a control. Functional maps resulting from comparison of the motor tasks with REST reveal activation in primary sensorimotor cortex, medial and lateral premotor areas, cingulate cortex, and parietal cortex. The task activation level, defined as the percentage MR signal increase for each task relative to REST, and the differential activation, defined as the percentage MR signal increase for RAND and SELF relative to FIX, were calculated in each area. All areas showed a higher activation level for RAND and SELF than for FIX. A significant difference in activation level or differential activation between SELF and RAND was found in the posterior part of the superior frontal sulcus, in a part of the premotor cortex on the lateral brain surface, in the anterior cingulate motor cortex, and in the posterior part of the superior parietal cortex. The high-resolution and single-subject approach, provided by fMRI, allowed the distinguishing of multiple foci in medial frontal areas, premotor cortex, and parietal cortex, reflecting the functional heterogeneity of these areas suggested by previous studies. PMID- 9344823 TI - Simultaneous measurement of DeltaR2 and DeltaR2* in cat brain during hypoxia and hypercapnia. AB - One of the most important issues in blood-oxygen-level-dependent (BOLD)-based brain functional magnetic resonance imaging is the understanding of the vascular structures that are responsible for the signal changes observed. The T2*-related signal changes observed during variations in susceptibility-induced magnetic field gradients are a function both of non-refocusable mechanisms, such as diffusion, and of refocusable effects such as field inhomogeneities. Conversely, T2-related signal changes are only a function of non-refocusable effects. It has been suggested that T2-weighted images could be less sensitive to blood susceptibility changes in a macrovascular environment than T2*-weighted images and could thus be more accurate in identifying the "activation" of the parenchyma rather than "draining vein" effects. In this study we use hypoxia and hypercapnia challenges in cats to provide a change in blood deoxyhemoglobin concentration (as a model for classic BOLD changes and not as a model for neuronal activation). A combined gradient echo and spin echo echo-planar-imaging (EPI) pulse sequence was used to map DeltaR2 (i.e., Delta(1/T2)) and DeltaR2* (i.e., Delta(1/T2*)) changes during the challenges. Our experiments demonstrate that: (i) the acquisition of T2-weighted EPI data does not in itself differentiate signal changes in the parenchyma from those occurring in regions around larger vessels, but that (ii) the simultaneous acquisition of T2- and T2*-weighted images could be useful in identifying microvascular regions in gray matter by analyzing the ratio DeltaR2/DeltaR2*. This value seems independent of the degree of deoxyhemoglobin concentration change, but is related to properties of the vascular environment. We suggest a possible application of the results to the study of brain function in humans. PMID- 9344824 TI - Involvement of primary motor cortex in motor imagery: a neuromagnetic study. AB - Functional brain imaging studies have indicated that several cortical and subcortical areas active during actual motor performance are also active during imagination or mental rehearsal of movements. Recent evidence shows that the primary motor cortex may also be involved in motor imagery. Using whole-scalp magnetoencephalography, we monitored spontaneous and evoked activity of the somatomotor cortex after right median nerve stimuli in seven healthy right-handed subjects while they kinesthetically imagined or actually executed continuous finger movements. Manipulatory finger movements abolished the poststimulus 20-Hz activity of the motor cortex and markedly affected the somatosensory evoked response. Imagination of manipulatory finger movements attenuated the 20-Hz activity by 27% with respect to the rest level but had no effect on the somatosensory response. Slight constant stretching of the fingers suppressed the 20-Hz activity less than motor imagery. The smallest possible, kinesthetically just perceivable finger movements resulted in slightly stronger attenuation of 20 Hz activity than motor imagery did. The effects were observed in both hemispheres but predominantly contralateral to the performing hand. The attempt to execute manipulatory finger movements under experimentally induced ischemia causing paralysis of the hand also strongly suppressed 20-Hz activity but did not affect the somatosensory evoked response. The results indicate that the primary motor cortex is involved in motor imagery. Both imaginative and executive motor tasks appear to utilize the cortical circuitry generating the somatomotor 20-Hz signal. PMID- 9344825 TI - Multimodal image coregistration and partitioning--a unified framework. AB - This paper presents a method for the coregistration and partitioning (i.e., tissue segmentation) of brain images that have been acquired in different modalities. The basic idea is that instead of matching two images directly, one performs intermediate within-modality registrations to two template images that are already in register. One can use a least-squares minimization to determine the affine transformations that map between the templates and the images. By incorporating suitable constraints, a rigid body transformation which directly maps between the images can be extracted from these more general affine transformations. A further refinement capitalizes on the implicit normalization of both images into a standard space. This facilitates segmentation or partitioning of both original images into homologous tissue classifications. Once partitioned, the partitions can be jointly matched, further increasing the accuracy of the coregistration. In short, these techniques reduce the between modality problem to a series of simpler within-modality problems. These methods are relatively robust, address a number of problems in image transformations, and require no manual intervention. PMID- 9344826 TI - Psychophysiological and modulatory interactions in neuroimaging. AB - In this paper we introduce the idea of explaining responses, in one cortical area, in terms of an interaction between the influence of another area and some experimental (sensory or task-related) parameter. We refer to these effects as psychophysiological interactions and relate them to interactions based solely on experimental factors (i.e., psychological interactions), in factorial designs, and interactions among neurophysiological measurements (i.e., physiological interactions). We have framed psychophysiological interactions in terms of functional integration by noting that the degree to which the activity in one area can be predicted, on the basis of activity in another, corresponds to the contribution of the second to the first, where this contribution can be related to effective connectivity. A psychophysiological interaction means that the contribution of one area to another changes significantly with the experimental or psychological context. Alternatively these interactions can be thought of as a contribution-dependent change in regional responses to an experimental or psychological factor. In other words the contribution can be thought of as modulating the responses elicited by a particular stimulus or psychological process. The potential importance of this approach lies in (i) conferring a degree of functional specificity on this aspect of effective connectivity and (ii) providing a model of modulation, where the contribution from a distal area can be considered to modulate responses to the psychological or stimulus-specific factor defining the interaction. Although distinct in neurobiological terms, these are equivalent perspectives on the same underlying interaction. We illustrate these points using a functional magnetic resonance imaging study of attention to visual motion and a position emission tomography study of visual priming. We focus on interactions among extrastriate, inferotemporal, and posterior parietal regions during visual processing, under different attentional and perceptual conditions. PMID- 9344827 TI - Editorial AB - No abstract Copyright 1997 Academic Press Limited PMID- 9344828 TI - Molecular mechanisms of glucocorticoid action in asthma. PMID- 9344829 TI - Recombinant human DNase (rhDNase) influences phospholipid composition, surface activity, rheology and consecutively clearance indices of cystic fibrosis sputum. AB - Cystic fibrosis (CF) is caused by mutation in the gene for the CF transmembrane conductance regulator which leads to massive, abnormally viscous, purulent sputum, chronic destructive endobronchitis and early death. Purified recombinant human (rh) DNase can digest extracellular DNA and its inhalation in these patients significantly improves lung function. To evaluate the poorly understood mechanisms, saliva protected sputum from patients treated with and without rhDNase were evaluated. Therapy with rhDNase resulted in a soluble sputum fraction that had a higher percentage of phosphatidylethanolamine, a phospholipid present mainly in cellular membranes, a much lower percentage of phosphatidylcholine, but a higher surface activity. The rigidity was significantly lower and the ratio of viscosity in proportion to elasticity increased. All these data are consistent with an increased clearability of the sputum by coughing, but not by mucociliary activity. Thus the interaction of inhaled rhDNase with the purulent mucus and the endobronchial inflammatory processes may induce changes that result in rheological properties favoring clearance of sputum by cough. PMID- 9344830 TI - Functional characterization of receptors for cysteinyl-leukotrienes in sheep trachealis muscle. AB - Cysteinyl-leukotrienes (CysLTs: LTC4, LTD4 and LTE4) are inflammatory mediators which significantly contribute to the airway obstruction in asthma. At least two distinct receptor subtypes exist for cysteinyl-leukotrienes, the CysLT1- and CysLT2-receptor. The purpose of this study was to test whether sheep trachealis muscle is a useful preparation for further characterization of CysLT2-receptors, previously implicated in contraction of human pulmonary veins. Leukotriene C4 and leukotriene D4 evoked contractile responses, leukotriene C4 being significantly more potent than leukotriene D4, whereas leukotriene E4 failed to elicit contractions. The response to leukotriene C4 exhibited tachyphylaxis upon repeated administration. There were no significant effects of epithelial denudation, NO-synthesis inhibition (L-NAME) or cyclooxygenase inhibition (indomethacin) on the responses to cysteinyl-leukotrienes or cholinergic agonists. Neither was responsiveness to different agonists changed by overnight storage. The responses to leukotriene C4 and leukotriene D4 were markedly potentiated when their metabolism was inhibited by S-hexyl glutathione and L cysteine. The selective CysLT1-antagonist ICI 198,615 had no significant effect on these responses. However, the combined CysLT1- and CysLT2-antagonist BAY u9773 competitively antagonized leukotriene C4 and leukotriene D4 (pA2 values of 7.0 and 6.8 against leukotriene C4 and leukotriene D4, respectively). The findings support that leukotriene C4 and leukotriene D4 act predominantly on CysLT2 receptors in sheep trachea. PMID- 9344831 TI - The effect of hydrogen peroxide on hypoxia, prostaglandin F2 alpha and potassium chloride induced contractions in isolated rat pulmonary arteries. AB - We have investigated the action of the product of the enzyme NADPH oxidase; hydrogen peroxide (H2O2), on the first phase of the hypoxic contraction, prostaglandin F2 alpha (PGF2 alpha)-induced contractions and potassium chloride (KCl)-induced contractions, in isolated rat pulmonary arteries in a wire myograph. Both concentrations of H2O2 (0.03 and 0.5 mM) produced initial contractions, and the higher concentration of H2O2 produced a significant inhibition of both the priming concentration of PGF2 alpha (5 microM) and the hypoxic contraction (P < 0.01 for both contractions). These effects were shown to be reversible, with contractions of a similar size to control values being seen to both PGF2 alpha (5 microM) and hypoxia following washout of H2O2 (P > 0.1 for both contractions). H2O2 (0.03 mM) was shown to have no significant effect upon either contraction (P > 0.1 for both contractions). H2O2 (0.5 mM) was also shown to have a significant inhibitory effect upon the efficacy (Emax) of the PGF2 alpha and KCl concentration-response curves (P < 0.01 for both contractions). This inhibition was again shown to be reversible. The higher concentration of H2O2 (0.5 mM) is clearly shown to be having a dual action, producing an initial contraction followed by inhibition of contractions to both PGF2 alpha and KCl. The mechanism by which H2O2 produces vasoconstriction is unclear, but it is suggested that H2O2 may inhibit the release of Ca2+ ions from intracellular stores as this is a common link between the modes of action of these two contractile agents. In addition to this, as an elevation in intracellular Ca2+ from intracellular stores appears to be a prerequisite for hypoxic pulmonary vasoconstriction (HPV), then this apparent mode of action of H2O2 could play an important role in the regulation of HPV and suggests a possible role for NADPH oxidase or a similar oxidoreductase as an oxygen sensor. PMID- 9344832 TI - Salmeterol inhibits the allergen-induced mononuclear cell proliferation and downregulates GM-CSF release and HLA-DR expression by monocytes. AB - beta 2-adrenoceptor agonists are known to attenuate several functions of human mononuclear cells in response to activation. Since monocytes and lymphocytes play a major pathogenetic role in allergic asthma, this study was designed to evaluate the hypothesis that salmeterol, a beta 2-adrenoceptor agonist with a long duration of action, could inhibit in vitro the allergen-induced activation of blood mononuclear cells (BMCs). BMCs were collected from subjects sensitized to Dermatophagoides pteronyssinus (Der p) and incubated with a purified Der p allergen extract to evaluate the ability of salmeterol to downregulate; (a) BMC proliferation; (b) IL-2 receptors (r) and HLA-DR surface antigen (ag) expression; (c) cytokine release. Der p induced a significant BMC proliferation (P < 0.01), associated with increased expression of HLA-DRag and IL-2r (P < 0.05) and with enhanced release of IL-2, GM-CSF, IL-1 beta, TNF-alpha and IFN-gamma (P < 0.01, each comparison). Salmeterol (10(-8)-10(-6) M) significantly inhibited, in a dose dependent manner, the Der p-induced BMC proliferation, reducing (at 10(-7) M) HLA DRag expression on monocytes and GM-CSF release (P < 0.05, each comparison). These data demonstrate that beta 2-adrenoceptor-mediated suppression of allergen induced BMC activation is associated with inhibition of cytokine release and of surface molecule expression, which are involved in the interaction between T lymphocytes and antigen-presenting cells. PMID- 9344833 TI - Changing the oxygen tension alters the ability of bronchodilators to protect against methacholine-induced challenge in bovine isolated bronchial rings. AB - The ability of atrial natriuretic peptide, salbutamol, sodium nitroprusside and isosorbide dinitrate to protect against challenge with methacholine in bovine isolated bronchi was compared in different O2 tensions. Perfusing the Krebs Henseleit solution with gas mixtures containing 95% O2 (hyperoxia), 20% O2 (approximately normoxia) and 0% O2 (hypoxia) produced O2 tensions in the organ baths of 524, 147 and 26 mm Hg, respectively. In hyperoxia, pre-incubation of atrial natriuretic peptide at concentrations of 3 x 10(-7) M and 10(-6) M significantly attenuated responses to methacholine, whereas in normoxia, these concentrations of atrial natriuretic peptide had no effect. Furthermore, in hypoxia, 3 x 10(-7) M and 10(-6) M atrial natriuretic peptide significantly enhanced responses to methacholine. Salbutamol, at concentrations of 3 x 10(-7) M and 10(-6) M significantly attenuated responses to methacholine in hyperoxia, whereas in normoxia and hypoxia, pre-incubation of salbutamol did not alter the methacholine response. Pre-incubation of 10(-5) M sodium nitroprusside significantly attenuated methacholine-induced contractions in hyperoxia and when the oxygen tension in the gas mixture was lowered to 20% or 0%, the ability of sodium nitroprusside to protect against methacholine challenge was enhanced. In hyperoxia, isosorbide dinitrate, at the 10(-4) M level, evoked a rightward shift of the methacholine response curve. Lowering the oxygen tension to either 20% or 0% enhanced the protectant effect of isosorbide dinitrate, with the effect being greater in 20% O2. Thus, the effect of these bronchodilators on methacholine induced challenge in hyperoxia O2 differed from those in normoxia and hypoxia, although the direction of the changes varied among the agents used. This suggests that the responses evoked by bronchodilators in 95% O2 may not necessarily predict those in the physiological range of oxygen tensions and that the relative effectiveness of bronchodilators may vary between normoxic and hypoxic conditions. PMID- 9344834 TI - Colchicine does not ameliorate bleomycin-induced pulmonary injury in hamsters. AB - We have evaluated the effect of colchicine, a potential antifibrotic drug, on bleomycin-induced pulmonary inflammation in hamsters. Pulmonary injury was induced by a single intratracheal (IT) instillation of bleomycin (bleo). Four groups of male Syrian hamsters each received one of four treatments: (1) IT bleo and twice daily intraperitoneal (IP) injections of colchicine (col) starting one day before IT instillation of bleo (bleo-col); (2) IT bleo and IP injections of saline (bleo-sal); (3) IT saline and IP colchicine (sal-col); and (4) IT saline and IP saline (sal-sal). Animals were sacrificed 28 days after IT treatment. Lung injury was evaluated histologically, biochemically and by analysis of bronchoalveolar lavage (BAL) fluid. Treatment of hamsters with colchicine did not ameliorate the bleo-induced lung injury, as determined by a semiquantitative morphological index that assesses the severity and extent of the lung injury on a scale of 0-3. Lung hydroxyproline measurements and BAL fluid cell count were also similar in bleo-col compared to bleo-sal hamsters. Colchicine did not prevent the bleo-induced restriction expressed by volume displacement. These results indicate that colchicine does not ameliorate the bleo-induced lung inflammation and fibrosis and call for further controlled investigations to justify the use of this drug in pulmonary disorders. PMID- 9344835 TI - The role of calcium in the regulation of apoptosis. AB - Apoptosis (programmed cell death) has gained widespread attention due to its roles in a variety of physiological and pathological processes, yet precisely how apoptosis is regulated by external and internal cues remains unclear. Work from our laboratories and others has implicated alterations in intracellular Ca2+ in apoptosis, and more recent work has defined particular biochemical processes that are targeted by Ca2+ in apoptotic cells. This review will summarize the role of Ca2+ in apoptosis within the context of what is known about the core components of the effector machinery for apoptosis. PMID- 9344836 TI - Benzoquinones inhibit the expression of inducible nitric oxide synthase gene. AB - Benzoquinones inhibited the production of NO induced by LPS and gamma-IFN in C6 glia cells. The mechanistic studies showed that benzoquinones inhibited the expression of iNOS mRNA through inactivation of NF kappa B. Benzoquinones may be a useful candidate for the development of a drug to treat disease due to iNOS gene over-expression. PMID- 9344837 TI - Up-regulation of integrin beta 3 expression by cyclic stretch in human umbilical endothelial cells. AB - The effect of uni-axial cyclic mechanical stretch on the expression of the adhesion protein integrin was investigated. Human umbilical endothelial cells (HUVECs) cultured on fibronectin coated silicon membranes were subjected to uni axial cyclic stretch. The level of expression of integrin beta 3 mRNA was found to be increased and peaked at 4 hours in response to cyclic stretch using a semiquantitative RT-PCR method. The increased level of the integrin mRNA from stretched HUVECs remained higher than that from non-stretched controls. The amount of integrin beta 3 also increased and peaked at 12 hr. Immuno-fluorescent microscopy revealed that the amount of integrin beta 3 adhesions increased in stretched HUVECs compared with that in non-stretched HUVECs. These results suggest that uni-axial cyclic stretch up-regulates the expression of integrin beta 3. This increase in integrin beta 3 may enhance the adhesiveness to the substratum and contribute to the protection of HUVECs against being peeled off from the vessel wall. PMID- 9344838 TI - On the regulation and function of human polo-like kinase 1 (PLK1): effects of overexpression on cell cycle progression. AB - The human protein kinase Plk1, a member of the polo-like kinase family, is known to function at mitosis. Here we show that the relative specific activity of Plk1 increases in mitosis, that Plk1 is specifically phosphorylated during mitosis, and that phosphatase treatment reduces mitotic Plk1 activity to interphase levels. To identify domains involved in the regulation of Plk1 activity, deletion mutants of Plk1 were constructed and their activities examined. Deletion of the extreme C-terminus of Plk1 substantially increased kinase activity, indicating that the C-terminus harbors an inhibitory domain. Finally, the consequences of over-production of wild-type and mutant Plk1 protein were analyzed, using transient transfection assays. Cells overexpressing Plk1 protein were able to enter mitosis and establish an apparently normal bipolar spindle. In contrast, progression through mitosis was transiently delayed, and cytokinesis appeared to be disturbed, as reflected by a significant increase in large cells with multiple, often fragmented nuclei. These results are relevant to recently proposed roles for Plks during both entry into and exit from mitosis. PMID- 9344839 TI - Molecular cloning of a novel brain-specific serine protease with a kringle-like structure and three scavenger receptor cysteine-rich motifs. AB - In order to find serine proteases specifically expressed in brain, we designed degenerate mixed primers for consensus sequences of serine protease domains. By PCR utilizing the primers, we have cloned a novel sequence from reverse transcripts of total RNA of mouse brain and used it as a probe to screen a mouse brain cDNA library. Overlapping cDNAs encoding a precursor of a novel brain specific serine protease (BSSP-3) were cloned. DNA insert of the longest clone consisted of 2614-bp with an entire open reading frame encoding a secretory/membrane-anchored precursor protein consisting of 761 amino acids (AA) which may be processed to yield an active enzyme of 245 AA. As found in known serine proteases, BSSP-3 enzyme domain contained a catalytic triad which consists of AA residues essential for the enzyme activity. In the upstream region of the enzyme domain that resides at C-terminus of the precursor protein, there are, from N-terminus to downstream, a sequence similar to a kringle structure and three repetitive ones highly similar to the scavenger receptor cysteine-rich (SRCR) motifs. Northern blot analysis demonstrated that mBSSP-3 mRNA was specifically expressed in the mouse brain, lung and kidney. We concluded that a novel brain serine protease, BSSP-3, is a new member of kringle and SRCR superfamilies. PMID- 9344840 TI - Nucleotide sequence and genetic complementation analysis of lep from Azotobacter vinelandii. AB - The lep of Azotobacter vinelandii is an 852-base-pair open reading frame (ORF) which encodes a protein of 284 amino acid residues. The translated protein shares 75% homology with leader peptidase I isolated from Pseudomonas fluorescens and 37% homology with leader peptidase I isolated from Escherichia coli. Five highly conserved regions found in the family of leader peptidase I proteins are conserved in A. vinelandii Lep. The putative membrane topology of the protein seems similar to that of E. coli leader peptidase I based on the hydrophobicity analysis of the predicted amino acid sequence. Southern blotting analysis of the A. vinelandii chromosome by probing with lep specific DNA revealed that lep is present as a single copy per the chromosome. A multicopy plasmid carrying A. vinelandii lep could complement a temperature sensitive lep mutant of E. coli strain IT41, suggesting that we have identified the functional copy of lep present on A. vinelandii genome. PMID- 9344841 TI - IFN-gamma upregulates anti-apoptotic gene expression and inhibits apoptosis in IL 3-dependent hematopoietic cells. AB - IFN-gamma is a cytokine which functions in a wide range of biological activities by inducing a number of early and delayed genes. In murine IL-3-dependent cell lines BAF-B03 and 32D, IFN-gamma upregulated bag-1 and bcl-xL gene expression. These cells revealed prolonged cell survival against IL-3-deprivation by IFN gamma stimulation. In contrast, human myeloma cell line RPMI8226, despite expression of IFN-gamma receptor, showed neither induction of their expressions nor prolonged cell survival against serum starvation-induced apoptosis by IFN gamma stimulation. Gene-transfer-mediated overexpression of BAG-1 protein in BAF B03 cells led to prolonged cell survival against IL-3-deprived apoptosis compared with control BAF-B03 transfectants, indicating that levels of BAG-1 expression are crucial for cell survival in BAF-B03 cells. Taken together, these studies suggest that induction of anti-apoptotic gene expression is a crucial factor for the anti-apoptotic function of IFN-gamma in IL-3-dependent immature hematopoietic cells. PMID- 9344842 TI - Mutational analysis reveals that an array of GCAAG/CTTGC motifs between sprit promoter sequences for RNA polymerase III is essential for neural BC1 RNA transcription. AB - BC1 RNA is expressed from an identifier (ID) sequence by RNA polymerase III (Pol III) and occurs in neural cells as a ribonucleoprotein particle (BC1 RNP). On the BC1 RNA gene, between the Pol III promoter A and B boxes, there is a region which contains short inverted repeats, including three GCAAG/CTTGC motifs. We found that a nuclear protein binds specifically to this region and, using an in vitro transcription system, demonstrated that point mutations within these motifs markedly inhibit BC1 RNA transcription. These results suggest that the GCAAG/CTTGC motif region and its binding protein may play a role in the transcription of BC1 RNA. Moreover, we demonstrated that transcription is repressed by a concomitant molar excess of BC1 RNA and that the BC1 RNA transcribed by this system forms an RNP with nuclear protein(s), suggesting some interaction of BC1 RNA with transcription factor(s). PMID- 9344843 TI - Erythropoietin and IL-3 induce tyrosine phosphorylation of CrkL and its association with Shc, SHP-2, and Cbl in hematopoietic cells. AB - The present study demonstrates that erythropoietin (Epo) and IL-3 induce tyrosine phosphorylation of the SH2/SH3-containing adapter protein CrkL and its transient association with tyrosine-phosphorylated SHP-2, Shc, and Cbl in a murine IL-3 dependent cell line, 32D, expressing the Epo receptor (EpoR). In these cells, CrkL was constitutively complexed with the guanine nucleotide exchange factor C3G, which was found to coimmunoprecipitate with Shc from Epo- or IL-3-stimulated cells. Studies using cells expressing mutant EpoRs showed that the Epo-induced tyrosine phosphorylation of CrkL is dependent on the membrane-proximal EpoR cytoplasmic region involved in the activation of Jak2 as well as the C-terminal 145 amino acid region which is required for tyrosine phosphorylation of SHP-2 and Shc. It was further revealed that CrkL is recruited to the tyrosine phosphorylated EpoR, most likely through its interaction with tyrosine phosphorylated Shc and SHP-2. These results suggest that CrkL is involved in the signaling pathways from the receptors for Epo and IL-3, most likely by modulating the activity of the Ras family GTPases through its interaction with C3G. PMID- 9344844 TI - The roles of valine 208 and histidine 211 in ligand binding and receptor function of the ovine Mel1a beta melatonin receptor. AB - Site-directed mutagenesis was used to study two residues, valine 208 and histidine 211, in transmembrane domain 5 of the ovine Mel1a beta melatonin receptor. A series of 4 mutants were constructed (V208A, V208L, H211F, H211L), and each engineered to contain a FLAG-epitope. Immunocytochemistry demonstrated that all the mutants were expressed in COS-7 cells at levels comparable to the FLAG-epitope tagged wild-type Mel1a beta receptor (approximately 120 fmol/mg protein). Ligand binding revealed however that all mutants had reduced affinities for 2-[125I]-iodomelatonin (Kd wild-type 139 pM, Kd mutants 320 to 989 pM). Competition studies, with a series of melatonin analogues, identified a probable interaction between histidine 211 and the 5-methoxy group of melatonin. The wild type receptor and both valine 208 mutants displayed a dose-dependent melatonin mediated inhibition of cyclic AMP levels in HEK293 cells, with IC50 values in the same rank-order as their melatonin binding affinities. Both H211F and H211L, however, did not display any melatonin mediated effects and may suggest that histidine 211 is critical for melatonin mediated receptor activation. PMID- 9344845 TI - Inhibition of human breast epithelial HBL100 cell proliferation by a dextran derivative (CMDB7): interference with the FGF2 autocrine loop [corrected]. AB - Fibroblast growth factor 2 (FGF2) has been shown to be an autocrine growth factor in human breast epithelial cells HBL100. Here we studied the effects of one dextran derivative (CMDB7) on this autocrine loop. CMDB7 caused a dose-dependent decrease of HBL100 growth in serum-free medium. [3H]thymidine uptake in HBL100 cells and Balbc/3T3 cells by exogenous FGF2 was inhibited by CMDB7. Receptor binding assays with radio-iodinated FGF2, IGF1, EGF showed that CMDB7 only displaced the binding of 125I-FGF2 in a dose dependent manner. Scatchard analysis revealed that the presence of CMDB7 reduced 78% and 82 % FGF2 binding capacity to its high and low affinity receptors respectively without altering the affinites of binding sites. These results suggest that CMDB7 exert its antiproliferative action on HBL100 cells by interfering with FGF2 autocrine loop. PMID- 9344846 TI - Establishment of two morphologically distinct PC12 cell lines resistant to 25-OH cholesterol toxicity. AB - Two novel populations of spontaneous PC12 cell mutants resistant to a toxic concentration of 25-OH-cholesterol (5 microg/ml, 12.5 microM) were isolated and designated as R25R and F25R based on cell morphology. R25R consisted of round cells that were morphologically similar to the parent PC12 cells, and responded to nerve growth factor by extending neurites. F25R was a group of process-bearing flat cells that did not assume a neuronal morphology in the presence of nerve growth factor. These two cell lines also acquired some cross-resistance toward other cholesterol oxides. Nerve growth factor induced prominent voltage-dependent calcium currents in parent PC12 cells and in R25R, but not in F25R. Further experiments indicated that the parent PC12 cells, R25R and F25R exhibited different properties when challenged with a variety of toxic insults, including amphotericin B, serum withdrawal and beta-amyloid protein treatment. PMID- 9344847 TI - Covalent glycoinositolphospholipid binding to hemoglobin: a new post translational modification of Hb occurring in hyperinsulinism with concomitant hypoglycemia. AB - In this work a novel hitherto unrecognised minor hemoglobin (Hb) fraction, which we detected previously in hemolysates of erythrocytes exposed to a high concentration of insulin under hypoglycemic conditions, both in vivo and in vitro, is analysed. The modification of Hb in HbA1x was shown to be due the addition of glycoinositolphospholipid (GPI) to the C termini of both beta polypeptide chains. A structurally related minor Hb fraction was identified in erythrocytes exposed in vitro to insulin-mimetic agent, trypsin. To our knowledge this is the first demonstration of such a modification of Hb, as well as the first demonstration of post-translational GPI binding to proteins in response to insulin. The mechanism proposed for GPI-Hb formation is briefly described. PMID- 9344848 TI - Cloning and characterization of novel CIS family genes. AB - We have reported two JAK-signaling modulators, CIS (cytokine-inducible SH2 protein) and JAB (JAK2 binding protein), which are structurally related. Here we cloned three additional CIS family genes (CIS2, CIS3, and CIS4) on the basis of an expression sequence tag (EST) database search. We also found at least two additional candidates of this gene family in the database. These genes were induced by erythropoietin and granulocyte-macrophage colony stimulating factor in certain hematopoietic cell lines. The SH2 domain and a C-terminal 40 amino acid region, designated the CIS homology domain (CH domain), are highly conserved in this family, while the N-terminal regions of these proteins share little similarity. A yeast two-hybrid assay and in vitro and in vivo binding assays revealed that in addition to JAB, CIS3 bound to the JAK2 tyrosine kinase domain (JH1), although the interaction of CIS3 with the JAK2-JH1 domain was much weaker than that of JAB. Transient expression of JAB and CIS3, but not other CISs, strongly inhibited leukemia inhibitory factor (LIF)-induced STAT3-reporter gene activation in 293 cells. Furthermore, constitutive overexpression of JAB and CIS3 in M1 leukemia cells prevented LIF-induced differentiation and growth arrest. Although the physiological function remains to be investigated, CIS family genes could play a role in the negative regulation of cytokine signaling by interacting with specific targets. PMID- 9344849 TI - Glucose-stimulated insulin secretion from rat islets of Langerhans is independent of mitogen-activated protein kinase activation. AB - The role played by mitogen-activated protein kinases (MAPKs) in the regulation of insulin secretion from adult rat islets of Langerhans was investigated by examining the effects of glucose, forskolin and 4beta phorbol myristate acetate (PMA) on islet MAPK activity and by measuring insulin secretion from islets in response to these agonists after inhibition of MAPK by PD 098059 (PD). Glucose (20mM) had a small (<2-fold) stimulatory effect on MAPK activity in isolated islets, and this was potentiated by forskolin (10 microM) and PMA (500nM), which also significantly stimulated MAPK activity at 2mM glucose. Pretreatment of islets with 50 microM PD inhibited MAPK activity, but had no effect on secretory responses to glucose, forskolin and PMA. These results suggest that although MAPK may be activated by insulin secretagogues in adult rodent islets, this can be dissociated from the exocytotic release of insulin. PMID- 9344850 TI - Identification of a novel Ca2+-stimulated S6-kinase in rat liver. AB - Extracellular calcium addition transiently stimulated two S6 peptide kinase activities in isolated rat hepatocytes. Mono Q chromatography revealed that the activities eluting at 0.15 M NaCl and 0.18 M NaCl were stimulated 4-fold and 2 fold, respectively. The kinase stimulated by calcium was a 40000-Mr S6 peptide kinase, as demonstrated by partial purification from whole liver. The protein kinase did not crossreact with antibodies directed against the N- or C-terminal part of p70 ribosomal S6 kinase (p70(S6K)) and the C-terminal part of p90 ribosomal S6 kinase (p90(rsk)). Following digestion of 40000-Mr S6 peptide kinase with trypsin, six peptides were sequenced. There was no similarity with the sequences of p70(S6K) and p90(rsk). Moreover, the obtained sequences could not be identified in the SwissProt or EMBL-genebank databases, suggesting that 40000-Mr S6 peptide kinase probably represents a novel protein kinase. PMID- 9344851 TI - Ascorbic acid is a stimulatory cofactor for mitochondrial glycerol-3-phosphate dehydrogenase. AB - Mitochondrial glycerol 3-phosphate dehydrogenase (mGPDH) from control and scorbutic guinea pig brain, liver and skeletal muscle and from normal rat liver was stimulated several fold by l-ascorbic acid (AA). The amount of AA that gave half-maximal stimulation of guinea pig brain mGPDH was 7.1 microM. At concentrations of AA higher than 500 microM, mGPDH activity decreased to nearly the same activity as in the absence of AA. D-Ascorbic acid, erythorbic acid, was equally potent in the activation of washed mitochondrial mGPDH activity. The AA activation of mGPDH was completely inhibited by 50 microM EGTA, but could be fully restored by the sequential addition of 100 microM Ca2+. The AA activation of mGPDH was likewise completely inhibited by iron specific chelators, bathophenanthrolinedisulfonic acid, desferrioxamine, and 1,10-phenanthroline, but the activation could not be restored by the addition of excess Ca2+. In the absence of AA, mGPDH activity was not inhibited by either EGTA or the iron chelators and Ca2+ addition had no effect on the activity. The iron-sulfur protein, succinic dehydrogenase (complex II), was not significantly different in brain mitochondria from control or scorbutic guinea pigs, and was not activated by the subsequent addition of AA. In the presence of AA, succinic dehydrogenase activity was not affected by either bathophenanthrolinedisulfonic acid or EGTA. The results suggest that mGPDH is a probable site of action of AA in the related glucose-coupled insulin release from pancreatic islets. PMID- 9344852 TI - Mice lacking the guanylyl cyclase C receptor are resistant to STa-induced intestinal secretion. AB - Heat-stable enterotoxin (STa) is an important causative agent of diarrheal disease throughout the world. STa is known to bind specifically to receptors in the intestine, provoking intense intestinal secretion. Binding of STa, or of the mammalian endogenous ligands guanylin and uroguanylin, activates the guanylyl cyclase C receptor (GC-C); the resulting elevation of cGMP levels stimulates chloride secretion via CFTR. We have generated knockout mice which completely lack the GC-C receptor. These mice are viable and show no obvious alteration in intestinal fluidity. However, GC-C null mice are refractory to the secretory action of STa, proving that the GC-C receptor is necessary for the diarrheal response induced by STa. PMID- 9344853 TI - 2,6-Bis((3,4-dihydroxyphenyl)-methylene)cyclohexanone (BDHPC)-induced apoptosis and p53-independent growth inhibition: synergism with genistein. AB - Estrogen binds to two classes of proteins in the cells, the high-affinity estrogen receptor (ER) as well as a low affinity estrogen type II binding site (EBS-II). Methyl p-hydroxyphenyllactate (MeHPLA) is an endogenous ligand for EBS II. Binding of MeHPLA to EBS-II has a growth regulatory effect in estrogen responsive cells, and levels of MeHPLA are decreased in breast cancer due to degradation by a specific esterase. 2,6-bis((3, 4-dihydroxyphenyl)-methylene) cyclohexanone (BDHPC) is an esterase-resistant analogue of MeHPLA which binds irreversibly to EBS-II and inhibits growth of breast cancer cells. In the present study, we analyzed the mechanism of growth inhibition by BDHPC. Treatment with BDHPC resulted in accumulation of cells in G1 phase and apoptosis. The G1 accumulation was not dependent on a functional p53 gene. The G1-specific growth inhibition by BDHPC was found to act synergistically with the G2/M-specific inhibition induced by the tyrosine kinase inhibitor genistein, suggesting this drug combination could be effectively used in cancer treatment. PMID- 9344854 TI - The sleep inducing factor oleamide is produced by mouse neuroblastoma cells. AB - Cis-9,10-octadecenoamide (oleamide) was isolated from the cerebrospinal fluid of sleep-deprived mammals and shown to induce sleep in rats. The enzyme catalyzing the hydrolysis of the amide bond of oleamide as well as of anandamide, the putative endogenous ligand of cannabinoid receptors, was purified from rat liver, cloned, shown to be expressed also in brain and named fatty acid amide hydrolase (FAAH). The enzymatic synthesis of oleamide from oleic acid and ammonia by rat brain microsomes has been also described. However, no evidence has been reported so far on the neuronal origin of oleamide, necessary in order to postulate for this compound a role as a neuromodulator. Here we show for the first time that oleamide is produced by a neuronal cell type and that its biosynthesis in intact neurons is not likely to occur through the direct condensation of oleic acid and ammonia. A lipid metabolite was extracted and purified from mouse neuroblastoma N18TG2 cells through a sequence of chromatographic steps and characterized as oleamide by means of gas chromatography/electron impact mass spectrometry (GC/EIMS). The amount of oleamide, as estimated by GC analyses carried out in comparison with known amounts of synthetic oleamide, was 55.0+/-09.5 pmols/10(7) cells, compared to less than 0.7 pmol/10(7) cells for anandamide in the same cells. When N18TG2 cells were prelabeled with [14C]oleic acid and the lipids extracted and purified, a radioactive component with the same chromatographic behavior as oleamide was found whose levels: (1) were not significantly influenced by stimulation with ionomycin; (2) were slightly increased by incubation with FAAH inhibitor phenyl-methyl-sulphonyl-fluoride (PMSF); (3) appeared to correlate with [14C]oleic acid incorporation into phospholipids but not with free [14C]oleic acid levels. N18TG2 cell membranes were shown to contain an enzymatic activity catalyzing the synthesis of oleamide from oleic acid and ammonia. This activity was inhibited by FAAH selective inhibitors arachidonoyltrifluoromethylketone and methylarachidonoylfluorophosphonate, as well as by an excess of anandamide, and by PMSF at the same concentration which increased oleamide formation in intact cells. These data suggest that a FAAH-like enzyme working "in reverse" may be responsible for the formation of oleamide in cell-free preparations but not in whole cells. PMID- 9344855 TI - Presenilin 1 binds to amyloid precursor protein directly. AB - Mutations in the presenilin genes are associated with early onset familial Alzheimer's disease and lead to accumulation of beta-amyloid peptide in the brain of patients, suggesting that presenilin abnormalities induce pathological processing of amyloid precursor protein (APP) in Alzheimer's disease. For the understanding of pathogenesis in this type of familal Alzheimer disease, it is important to know whether presenilins are directly involved in the metabolism of beta-amyloid or not. To test whether presenilin 1 (PS1) directly binds to APP, we performed two-hybrid interaction assays between these proteins in yeast cells by using bait plasmids for normal and mutant PS1 and prey plasmids for APP fragments corresponding to the different molecular portions. Positive interaction was observed in any combination between PS1 bait plasmids and APP prey plasmids. Therefore, our results show that PS1 binds to APP directly and suggest that the PS1 protein itself is involved in the metabolism of beta-amyloid peptide. PMID- 9344856 TI - Epidermal growth factor stimulates the tyrosine phosphorylation of SHPS-1 and association of SHPS-1 with SHP-2, a SH2 domain-containing protein tyrosine phosphatase. AB - SHPS-1 is a 120 kDa glycosylated receptor-like protein that contains immunoglobulin-like domains in its extracellular region and four potential tyrosine phosphorylation for SH2 domain binding sites in its cytoplasmic region. Epidermal growth factor (EGF) stimulated the rapid tyrosine phosphorylation of SHPS-1 and subsequent association of SHPS-1 with SHP-2, a protein tyrosine phosphatase containing SH2 domains, in Chinese hamster ovary cells overexpressing human EGF receptors. In the cells overexpressing SHPS-1, the tyrosine phosphorylation of SHPS-1 was more evident than that observed in parent cells. However, overexpression of SHPS-1 alone did not affect the activation of MAP kinase in response to EGF. These results suggest that SHPS-1 may be involved in the recruitment of SHP-2 from the cytosol to the plasma membrane in response to EGF. PMID- 9344857 TI - A novel ligand for an SH3 domain of the adaptor protein Nck bears an SH2 domain and nuclear signaling motifs. AB - Nck is a small protein composed of Src homology regions (SH) 2 and 3, paralleling the adaptors c-Crk and Grb2/Ash, but its function remains enigmatic. To clarify Nck signaling, a human brain cDNA library was searched for targets of the SH3 moiety of Nck. A novel molecule detected therefrom (referred to as Nck-, Ash- and phospholipase Cgamma-binding protein 4) contained proline-rich sequences and, through the function of one of them, interacted with the middle SH3 domain of Nck. A NAP4 fusion peptide exhibited an affinity for Nck, Ash and phospholipase Cgamma in whole cell lysates. NAP4 also had an SH2 domain, which could bind to activated EGF receptor. These intermolecular interactions imply the intricacy of Nck-mediated signaling around the receptor protein-tyrosine kinases. In addition, NAP4 bore a putative nuclear localization signal and a Q-run/P-run composite, both characteristic of nuclear proteins, and might therefore relate to the presence of Nck in the cellular nucleus. PMID- 9344858 TI - c-Src activates both STAT1 and STAT3 in PDGF-stimulated NIH3T3 cells. AB - Treatment of cells with PDGF and EGF specifically induces STAT1 and STAT3, which became phosphorylated on tyrosine residues to form homo and heterodimers: in these configurations they translocate into the nucleus where they act as transcription activators. However little is known about the activation of STATs in growth factor receptor signal transduction. Recently it has been shown that v Src modulates the tyrosine phosphorylation of STAT3 but not of STAT1. Here we report that the cellular Src tyrosine kinase is involved in the activation of both STAT1 and STAT3 in PDGF stimulated NIH3T3 cells. Both tyrosine phosphorylation and DNA binding activity of STAT1 and STAT3 are up-regulated in c Src overexpressing cells, while we observe the opposite phenomenon in cells overexpressing the dominant negative Src. Furthermore, our results show that STAT1 co-immunoprecipitates with c-Src, suggesting that the activation of STATs by Src occurs via a direct interaction. Taken together, these data suggest that c Src is involved in activation of both STAT1 and 3 in PDGF signal transduction. PMID- 9344859 TI - Reconstitution of highly expressed human heart muscle carnitine palmitoyltransferase I. AB - The human heart muscle carnitine palmitoyltransferase I (M-CPTI) gene was expressed at high levels from a strain of the methylotrophic yeast Pichia pastoris containing approximately 24 copies of the expression vector. Levels of M CPTI were more than ten-fold higher than previously reported by our group with a single-copy strain (Arch. Biochem. Biophys., in press) and were sufficient to perform reconstitution studies on the membrane protein, a key step in purification and structural analysis of the enzyme. Solubilization of yeast mitochondria containing M-CPTI in 5% Triton X-100 abolished M-CPTI activity. The detergent-inactivated M-CPTI was then reconstituted by removal of the detergent in the presence of phospholipids. The reconstituted proteoliposomes exhibited M CPTI activity of 2.4 nmol palmitoylcarnitine formed/mg protein/min, a recovery of 23% of the activity present in the starting mitochondrial preparation. The malonyl-CoA sensitivity of the reconstituted reactivated M-CPTI was 88%. This is the first demonstration of direct reactivation of malonyl-CoA-sensitive M-CPTI activity from solubilized materials from any organism. Previously, M-CPTI was presumed to be irreversibly inactivated by detergents. PMID- 9344860 TI - Regulated expression of 5'-deleted mouse GLUT4 minigenes in transgenic mice: effects of exercise training and high-fat diet. AB - Fourteen kb murine GLUT4 minigene (= -7395 GLUT4) contains DNA sequence that confers tissue specific, exercise-induced up-regulation of the GLUT4 gene in skeletal muscle and high-fat diet induced-down-regulation in white adipose tissue. To identify the DNA sequences required for regulated expression, we generated GLUT4 minigene transgenic mice harboring 3237, 2000, 1000, and 442 bp of 5'-flanking region, all exons and introns, and 1 kb of 3'-flanking sequence of the mouse GLUT4 gene. The -3237-, -2000-, and -1000-GLUT4 constructs were expressed in a tissue-specific manner identical to the endogenous GLUT4. Exercise induced up-regulation and high-fat diet-induced down-regulation of these constructs also paralleled those of the endogenous GLUT4 gene. In contrast, the 442 GLUT4 construct was expressed substantially in skeletal muscle (gastrocnemius and quadriceps) and heart, but was only expressed very weakly in white adipose tissue and was not expressed in brown adipose tissue. Furthermore, this -442 GLUT4 construct failed to respond to exercise or a high-fat diet in either muscle or adipose tissue. These results indicate that brown and white adipocyte-specific enhancer(s) and exercise- and high-fat diet-responsive elements are located between bases -1000 and -442 of the murine GLUT4 5'-flanking region. PMID- 9344862 TI - Inhibition of lipopolysaccharide and cytokine mixture-mediated hepatocyte nitric oxide synthesis by dimethyl sulfoxide. AB - Treatment and pretreatment of hepatocytes with 2% dimethyl sulfoxide (DMSO) inhibited lipopolysaccharide and cytokine mixture (LPS/CM)-mediated NO synthesis in hepatocytes without any obvious effects on cell viability. DMSO at concentrations of 0.5-4% stimulated DNA replication and increased albumin secretion in LPS/CM-treated hepatocytes. Genisein, a inhibitor of protein tyrosine kinase (PTK), inhibited LPS/CM-mediated NO synthesis in hepatocytes. These results suggest that PTK is critical for hepatocyte NO synthesis, and DMSO inhibited NO synthesis may be associated with prevention of LPS/CM-induced PTK activation in hepatocytes. PMID- 9344861 TI - cDNA cloning of a human dishevelled DVL-3 gene, mapping to 3q27, and expression in human breast and colon carcinomas. AB - dishevelled (Dsh) is a member of the segment polarity gene family in Drosophila which plays an important role in the early developmental patterning processes. A human homologue of Dsh (DVL-1) has recently been described. Here, we report the cloning of a second human homologue of Dsh (called DVL-3) by cDNA library screening. The human DVL-3 gene encodes a predicted 716 amino acid protein which exhibits 98% amino acid identity with mouse Dvl-3 and 49% with Drosophila Dsh. DVL-3 was mapped to 3q27. The expression of DVL-3 mRNA was detected in 30 human cell lines and 2 primary cell cultures. DVL-3 mRNA was detected in normal human breast tissues (n = 4) and tumours (n = 25). Statistically, there was no difference in DVL-3 mRNA level between normal breast tissues and tumours. In human colorectal samples, DVL-3 was expressed equally in matched normal tissues, polyps and tumours. The data indicates that DVL-3 is widely expressed in human cells and supports the notion of a new developmental gene family for dishevelled which may have a widespread role in signal transduction. PMID- 9344863 TI - Amino acid residues in both the DNA-binding and ligand-binding domains influence transcriptional activity of the human peroxisome proliferator-activated receptor alpha. AB - We have investigated the basis of the lack of activity of a natural variant human peroxisome proliferator-activated receptor alpha, hPPARalpha6/29. A subcloning approach was used to change the four variant amino acids in the hPPARalpha6/29 sequence, individually and in combination, to those found in an active human PPARalpha. Individual amino acid "back mutations" were unable to confer on hPPARalpha6/29 the ability to be activated by peroxisome proliferators in a transient transfection assay. Although hPPARalpha6/29 was able to bind specifically to DNA in the presence of the retinoid X receptor alpha (RXRalpha), the complete restoration of receptor transcriptional activity required two separate back mutations of the hPPARalpha6/29 sequence, namely amino acid 123 in the DNA binding domain, and amino acid 444 close to the C-terminus. This suggests that sequences in the PPARalpha DNA binding domain influence other receptor functions besides DNA binding. PMID- 9344864 TI - Molecular cloning of cDNA and genomic DNA for human 25-hydroxyvitamin D3 1 alpha hydroxylase. AB - The 25-hydroxyvitamin D3 1 alpha-hydroxylase (1 alpha-hydroxylase) is a cytochrome P450 enzyme that catalyzes the conversion of 25-hydroxyvitamin D3 to 1 alpha,25-dihydroxyvitamin D3. This enzyme plays an important role in calcium homeostasis. Here we report the molecular cloning of cDNA and gene for human 1 alpha-hydroxylase. The cDNA clone was obtained from a human kidney cDNA library by cross-hybridization with a previously cloned rat cDNA probe. The cDNA consists of 2469 bp and encodes a protein of 508 amino acids that shows 82.5% sequence identity with the rat enzyme. A computer-aided homology search revealed that 1 alpha-hydroxylase shares a relatively high homology with vitamin D3 25 hydroxylase (about 40% amino acid identity). Northern blot analysis showed that the 2.5-kb mRNA is most abundant in kidney. The gene for human 1 alpha hydroxylase spans approximately 6 kb, is composed of nine exons, and is present as a single copy. This molecular cloning makes it possible to investigate the genetic mechanism of diseases related to calcium metabolism, including vitamin D dependency rickets type I. PMID- 9344865 TI - Overexpression, purification, and use of a soluble human interleukin-4 receptor alpha-chain/Ig gamma 1 fusion protein for ligand binding studies. Characterization of ligand binding to soluble IL-4 receptor alpha-chain by surface plasmon resonance measurements and by microtiter-plate-based ELISA with biotinylated IL-4. AB - The pleiotropic cytokine IL-4 transmits cellular signals mainly via the IL-4 receptor complex, with the alpha-chain as the high affinity binding subunit. Here we describe the overexpression of a soluble IL-4R alpha-chain (sIL-4R) as a fusion to immunoglobulin gamma 1 heavy chain, consisting of the H-CH2-CH3 domains, in baby hamster kidney cells. The dimeric fusion protein named sIL-4R:E gamma 1 was purified from culture supernatant by protein-A affinity chromatography, yielding up to 10 mg/l homogenous protein which was highly stable. The antibody-like features of the sIL-4R:E gamma 1 fusion protein allowed immobilization on a biosensor matrix for surface plasmon resonance measurements by direct amine coupling as well as immobilization on microtiter plates coated with protein A for displacement binding. Kinetic parameters (kon and koff) for binding of IL-4 or the antagonistic mutant IL-4(Y124D) to the sIL-4R:E gamma 1 fusion protein on the chip as determined with the BIAcore instrument showed a high affinity binding with KD = 239 +/- 35 pM and KD=148 +/- 33 pM, respectively. The extremely high kon rate and the relatively slow koff rate for both ligands highlighted the limits of the BIAcore technology. The binding affinity as calculated in displacement binding studies with biotinylated IL-4 was similar for IL-4 and IL-4(Y124D) (IC50=1.1nM), thus offering a simple alternative for initial characterization of IL-4 mutants. PMID- 9344866 TI - A cluster of four novel human G protein-coupled receptor genes occurring in close proximity to CD22 gene on chromosome 19q13.1. AB - In our search for novel human galanin receptor (GALR) subtypes, human genomic DNA was PCR amplified using sets of degenerate primers based on conserved sequences in human and rat GALR. The sequence of one of the subcloned PCR products revealed homology to a sequence in the 3' region of the human CD22 gene following a BLAST search of GenBank's database. A search for open reading frames (ORF) in the non coding CD22 sequence resulted in identification of two novel putative intronless genes, GPR40 and GPR41. The recent submission of sequence overlapping the downstream CD22 sequence revealed a possible polymorphic insert containing a third intronless gene, GPR42, sharing 98% amino acid identity with GPR41, followed by a fourth intronless gene, GPR43. Thus, the GPR40, GPR41, GPR42, and GPR43 genes, respectively, occur downstream from CD22, a gene previously localized on chromosome 19q13.1. The four putative novel human genes encode new members of the GPCR family and share little homology with GALR. PMID- 9344867 TI - Asp278 of human beta-adrenergic receptor kinase 1 is essential for phosphorylation activity. AB - Asp278 of beta-adrenergic receptor kinase 1 (betaARK1) was suggested to play a key role in substrate recognition of beta2-adrenergic receptors in our previous study, in which a three-dimensional model of betaARK1 was studied in comparison with a crystal structure of PKA-PKI5-24 complex. In the present study, to confirm the molecular recognition mechanism at Asp278 of betaARK1, two mutants of betaARK1, D278R and D278A, were designed based on molecular modeling studies and produced by Sf-9 cells. As predicted by the molecular modeling study, the mutants showed no kinase activities while wild type betaARK1 phosphorylated beta2 adrenergic receptors in a concentration-dependent manner. These results strongly suggest the involvement of Asp278 in substrate recognition by betaARK1. The results also suggest a high reliability of the three-dimensional model of betaARK1. PMID- 9344868 TI - High pressure modulation of Escherichia coli DNA gyrase activity. AB - Elevated pressures greater than 551 bar were found to inhibit the DNA supercoiling activity of Escherichia coli DNA gyrase. A large fraction of the inhibitory effect could be reproduced by preincubation of the enzyme at high pressure prior to enzymatic assay at 1 bar. It is proposed that elevated pressure influences gyrase structure, most likely by promoting the dissociation of its subunits, however, it is also possible that effects on enzyme activity exist. PMID- 9344869 TI - The inhibition of synthesis of a beta-chemokine, monocyte chemotactic protein-1 (MCP-1) by progesterone. AB - The control of chemokines in reproductive tissues has not been well characterised. Progesterone plays a major part in many reproductive processes and an interaction between progesterone and the immune system has been postulated. MCP-1 is a beta chemokine that attracts and activates macrophages, controls vascular smooth muscle cells, and can modulate T helper cell cytokine production. MCP-1 may also play a role in reproductive processes such as ovulation and parturition. MCP-1 synthesis is stimulated by the transcription factor NF-kappa B and is inhibited by glucocorticoid but the relevance of progesterone control in reproductive tissue is unknown. The effects of progesterone on the production in both choriodecidual cells and a breast cancer cell line T47D, which expresses an oestrogen insensitive progesterone receptor, were investigated. A synthetic progestin (medroxyprogesterone acetate) inhibits choriodecidual cell production of MCP-1; this inhibition was reversed by the antiprogestin RU486. MCP-1 release from T47D cells can be stimulated by IL-1 and this production is inhibited by progesterone with an ED50 of less than 10(-9) M. A glucocorticoid (dexamethasone) had no effect on MCP-1 release in this system, suggesting that glucocorticoid receptor-mediated responses were impaired under these conditions. These results demonstrate that an indirect effect of progesterone on the immune system is possible in reproductive tissues, whereby the initial effect of progesterone on epithelial or fibroblast cells would be transmitted to leukocytes. PMID- 9344870 TI - Stimulation of Ca2+ release-activated Ca2+ channels as a potential mechanism involved in non-genomic 1,25(OH)2-vitamin D3-induced Ca2+ entry in skeletal muscle cells. AB - As in other vitamin D target cells, activation of voltage-dependent Ca2+ channels (VDCC) mediates the fast, non-genomic, 1,25(OH)2D3 stimulation of Ca2+ influx in skeletal muscle cells (SMC). 1,25(OH)2D3 has also been shown to rapidly induce the release of Ins(1,4,5)P3 in SMC. Experiments were performed to investigate whether Ca2+ release-activated Ca2+ channels (CRAC) also participate in the mechanism by which 1,25(OH)2D3 regulates Ca2+ entry into these cells. In cultured chick SMC loaded with Fura-2/AM the hormone (10(-12) - 10(-8) M) induced a rapid (30 sec) followed by a sustained (up to 5 min) increase in intracellular Ca2+ concentration ([Ca2+]i) associated to Ca2+ mobilization from internal stores and influx of extracellular Ca2+, respectively. Thus, the initial, transient, 1,25(OH)2D3-dependent increment in [Ca2+]i could be observed in Ca2+-free medium and was abolished by the PLC inhibitor U73122. Readdition of Ca2+ to cells that had undergone the initial 1,25(OH)2D3-induced [Ca2+]i rise in Ca2+ free medium resulted in a fast increment in [Ca2+]i indicating the existence of a hormone activated CRAC entry pathway. The sustained phase of the Ca2+ response to 1,25(OH)2D3 was only partially (60%) suppressed by nifedipine, whereas lanthanum (10 microM) completely abolished the hormone effects. Accordingly, depletion of intracellular Ca2+ stores by thapsigargin reproduced 1,25(OH)2D3-induced Ca2+ influx, inhibiting any further response to the sterol. 1,25(OH)2D3 increased the rate of quenching of Fura-2 fluorescence by Mn2+, indicating activation of Mn2+ permeable channels. Altogether, these results provide the first evidence involving CRAC channels in the rapid modulation of Ca2+ entry in animal cells by 1,25(OH)2D3. PMID- 9344871 TI - Expression of c-myc is down-regulated as mouse mammary epithelial cells become confluent. AB - We have investigated the expression profile of c-myc in the mammary gland. During pregnancy when the gland is actively growing c-myc mRNA was present, while in the differentiated lactating gland no c-myc mRNA was detected. This correspondence between the differentiation state and c-myc mRNA levels in the mouse mammary gland in vivo was paralleled by HC11 mouse mammary epithelial cells in vitro. Firstly, the endogenous c-myc gene was suppressed in confluent compared to growing HC11 cells. In addition, treating the cells with lactogenic hormones did not induce c-myc expression. Secondly, a stably transfected c-myc-CAT reporter construct was similarly down-regulated. Furthermore, using this transfection model, we demonstrate that the mechanism(s) involved in regulating c-myc expression must act through the P1 and P2 core promoter and exon 1. Finally, we demonstrate that suppression of c-myc expression occurs when HC11 cells growth arrest as they become confluent. PMID- 9344872 TI - High-density lipoprotein from hypercholesterolemic animals has peroxidized lipids and oligomeric apolipoprotein A-I: its putative role in atherogenesis. AB - Oxidized lipoproteins have been involved in the pathogenesis of atherosclerosis and atherosclerotic lesions contain oxidized low density lipoprotein. Conversely, the presence of oxidized high density lipoprotein (HDL) in vivo has not been clearly established. Oxidation of HDL in vitro models produces an increase in peroxidized lipids and the appearance of apolipoprotein A-I (apo A-I) oligomers. We investigated the oxidative status of HDL in an in vivo model: the hypercholesterolemic chicken. The HDLs from control and hyperlipemic animals were analyzed for the content of lipid peroxides employing spectroscopic and fluorescence techniques, for the level of apo A-I oligomerization, and for susceptibility to in vitro oxidation. HDL from hypercholesterolemic chickens was more peroxidized (as detected by fluorescence), had higher amount of oligomeric apo A-I, and was oxidized to a greater extent by uv irradiation than that of control animals. We speculate that apo A-I oligomerization could be a key step in the atheroma formation. PMID- 9344873 TI - Comparison of ectopic osteoinduction in vivo by recombinant human BMP-2 and recombinant Xenopus BMP-4/7 heterodimer. AB - Two micrograms recombinant human bone morphogenetic protein-2 (rhBMP-2) and 2 microg recombinant Xenopus bone morphogenetic protein-4/7 heterodimer (rxBMP 4/7ht) were implanted into a calf muscle pouch in rats using atelopeptide type I collagen (CL) solution as the carrier. Three weeks later induced bone in the rhBMP-2 + CL group (group A; n=5) and the rxBMP-4/7ht + CL group (group B; n=5) was investigated radiologically and histologically. Bone trabeculum and marrow were induced in all cases of both groups. In group A the area of radioopaque oval shadows was 5.78 mm2, wider than 4.64 mm2 in group B. The radoopacity in group A was 74.6, higher than 57.6 in group B. Histometry of the microscopic views showed that the trabecular area was 0.92 mm2 in group A and 0.22 mm2 in group B, while the trabecular percentage occupied in the overall lump area was 18.07% in group A and 5.51% in group B. The trabecular form in group A was more massively coral than in group B. This study indicated that rhBMP-2 has a higher ectopic osteoinduction ability in vivo than rxBMP-4/7ht. PMID- 9344874 TI - Visualization of mitochondria with green fluorescent protein in cultured fibroblasts from patients with mitochondrial diseases. AB - cDNAs for green fluorescent protein (GFP) and for a GFP fusion protein containing the presequence of human ornithine transcarbamylase (pOTC-GFP) were transfected into cultured human fibroblasts. GFP cDNA gave diffuse fluorescence throughout the cytoplasm and the nucleus, whereas pOTC-GFP cDNA gave mitochondria-associated fluorescence. Fluorescent mitochondrial structures could be classified into five patterns: thread-like mitochondria, fine thread-like ones, rod-like ones, granular ones, and granular ones with weak cytosolic fluorescence. pOTC-GFP mutants resulted in a loss of mitochondrial fluorescence and an appearance of weak fluorescence throughout the cytoplasm. pOTC-GFP cDNA was transfected into fibroblasts from patients with various mitochondrial diseases. Higher ratios of fibroblasts with granular mitochondria and those with fine thread-like ones were observed in a patient with Reye's syndrome and a patient with Kearns-Sayre syndrome. Weak cytosolic fluorescence was sometimes observed in fibroblasts from these patients. This method will be useful to analyze mitochondrial structural alterations and disorders of mitochondrial protein import. PMID- 9344875 TI - Xenopus FK 506-binding protein homolog induces a secondary axis in frog embryos, which is inhibited by coexisting BMP 4 signaling. AB - FK 506-binding protein (FKBP) is an immunosuppressant mediator in mammals, but its endogenous physiological function has yet to be determined. Here we report a Xenopus homolog of FKBP, which is expressed at early stages of development. Injection of synthesized Xenopus FKBP mRNA, as well as murine constitutively active calcineurin, induced a secondary axis in Xenopus embryos, while an FKBP mutant which does not bind to calcineurin did not. This secondary-axis-inducing effect was inhibited when FKBP was coinjected with Xmad 1 or XBMP 4 mRNA. These results suggest that FKBP modifies BMP 4 signalling by recruiting calcineurin and may have an important role in axis formation during Xenopus development. PMID- 9344876 TI - Intracellular location of SNAP-25 in human neutrophils. AB - Exocytosis plays an essential role in the physiological functions of human neutrophils. Although SNAP-25 is considered to play a key role in vesicle membrane fusion, it has been detected almost exclusively in the neuronal system. Using different specific antibodies to SNAP-25, we have identified in the membrane fraction of resting human neutrophils an immunoreactive band with the same molecular mass observed in brain homogenates. Immunoblot analysis of subcellular fractions of neutrophils revealed that SNAP-25 protein was found in the granule membrane fraction, but not in the cytosolic and plasma membrane fractions. Granule localization for neutrophil SNAP-25 was further demonstrated by confocal and immunoelectron microscopy. Furthermore, SNAP-25 was mainly located in the morphologically defined neutrophil peroxidase-negative granules, which are mobilizable upon cell activation. In addition, the protein was specifically cleaved by botulinal neurotoxin A, as observed in brain homogenate. These findings reveal the presence of SNAP-25 in the granule membranes of human neutrophils. PMID- 9344877 TI - Isolation of the embryonic form of smooth muscle myosin heavy chain (SMemb/NMHC B) gene and characterization of its 5'-flanking region. AB - To examine the molecular mechanisms that regulate the expression of the SMemb/NMHC-B gene, a nonmuscle myosin heavy chain isoform predominantly expressed in fetal aorta, we have isolated and characterized the 5'-flanking region of the rabbit SMemb/NMHC-B gene. Transient transfection experiments demonstrated that 105 base pairs of 5'-flanking sequence was necessary to direct high level transcription in C2/2 cells, vascular smooth muscle cells derived from rabbit aorta. An essential cis-regulatory element was localized between -100 and -91 base pairs from the transcription start site based on the results that replacement mutagenesis within this region significantly reduced promoter activity. Sequence of this region is completely conserved between mouse and rabbit and fits no known DNA binding consensus. Gel mobility shift assays revealed that a specific DNA-protein complex was formed at this site with nuclear extracts from C2/2 cells, which can be competed by H-2Kb CCAAT box but not by Hsp70 CCAAT box or other CCAAT-containing sequences. We conclude that expression of the SMemb/NMHC-B gene is regulated through an interaction between a sequence element located at -100 and a distinct member of CCAAT-binding proteins. PMID- 9344878 TI - Polymorphic expression of multidrug resistance mRNA in lung parenchyma of nonpregnant and pregnant rats: a comparison to cystic fibrosis mRNA expression. AB - Multidrug resistance (MDR1b) and cystic fibrosis transmembrane conductance regulator (CFTR) proteins are members of the "ATP-binding cassette" superfamily of transporters. They are associated with chloride channel activities and ATP secretion and have complementary patterns of expression in several organs. In the rat uterus, CFTR expression is replaced by MDR1b expression during pregnancy. We have studied whether expression of MDR1b and CFTR also vary in the lung during pregnancy. No variations in MDR1b or CFTR mRNA levels during pregnancy were detected. However, there was an unusual degree of variation in MDR1b mRNA expression in lung parenchyma between animals in both the control group and the pregnant group. If present among humans, polymorphic expression of MDR1 in lung parenchyma may explain part of the differences in lung symptomatology observed in the CF patients carrying the same mutation. PMID- 9344879 TI - Rab11 is associated with transferrin-containing recycling compartments in K562 cells. AB - 3'RACE PCR was used to survey Rab transcripts synthesized by the human hematopoietic K562 cell line. Among the identified GTP-binding proteins, Rab11 was discovered. This result was unexpected since Rab11 previously had been found associated with polarized and secretory cells. Rab11 mRNA was abundant compared to that for other Rabs in K562 cells; protein levels represented 0.05-0.1% of total membrane protein. Localization of Rab11 using confocal immunofluoresence microscopy revealed extensive overlap with transferrin in recycling and/or exocytic compartments and suggests that Rab11 in non-polarized and non-secretory cells may play a role in the trafficking and recycling of internalized proteins. PMID- 9344880 TI - The tumor suppressor p53 is a negative regulator of estrogen receptor signaling pathways. AB - The estrogen receptor (ER) is a ligand-dependent transcription factor which regulates growth, development, differentiation and reproduction. To test the hypothesis that the diverse effects of the ER could be mediated by interacting with other transcription factors/oncogenes, the present study assessed its interaction with the tumor suppressor p53. p53 is a transcription factor which is involved in cell cycle regulation and apoptosis. We found that the wild-type p53 physically interacted with ER in vivo and repressed the estrogen-activated transcriptional activity. However, p53 mutants had no or reduced repression effect, depending on the sites of mutation. These findings suggest that p53 can cross talk with the ER in hormone-activated signaling pathways in cells. PMID- 9344881 TI - Beta 2-microglobulin induces stromelysin production by human synovial fibroblasts. AB - beta 2-Microglobulin (beta 2-m) is a major constituent of amyloid fibrils in hemodialysis-associated amyloidosis (HAA), a serious complication in patients on long-term hemodialysis. The most distinctive pathological feature of HAA is the deposition of amyloid fibrils with subsequent articular inflammation and destruction. However, the pathological role of beta 2-m is not well known at present. We investigated the effects of beta 2-m on the production of proteinases from synovial fibroblasts isolated from patients with rheumatoid arthritis. beta 2-m stimulated synovial fibroblasts to produce stromelysin, a neutral matrix metalloproteinase (MMP-3). The production of MMP-2 and of a tissue inhibitor of metalloproteinase-1 (TIMP-1) were not enhanced by beta 2-m-treated synovial fibroblasts. Stromelysin is capable of degrading several components of the extracellular matrix and believed to be the key enzyme causing articular destruction in inflammatory joint diseases. Our results suggest a novel role for beta 2-m in articular inflammation and destruction mediated by stromelysin in HAA. PMID- 9344882 TI - Apoptosis in gastric epithelial cells is induced by Helicobacter pylori and accompanied by increased expression of BAK. AB - Carriage of the bacterium H. pylori in the human stomach is associated with evidence of increased epithelial cell apoptosis. This may be of significance in the etiology of gastritis, peptic ulcers, and neoplasia. The ability of H. pylori to directly induce epithelial apoptosis was examined in vitro by fluorescence and electron microscopy, flow cytometry, and DNA fragmentation ELISA. The induction of apoptosis by H. pylori was time and concentration-dependent and inhibited by preventing direct bacterial-epithelial cell contact. Apoptosis was accompanied by increased expression of Bak, with little change in expression of other Bcl-2 family proteins. The expression of Bak was also increased in gastric biopsies from patients colonized by H. pylori. Thus, H. pylori induces gastric epithelial cell apoptosis, by a Bak-dependent pathway. PMID- 9344883 TI - Up-regulation of angiotensin type 2 receptor mRNA by angiotensin II in rat cortical cells. AB - The present experiment demonstrates that the exposure of angiotensin II (AII) produced an up-regulation of the AT2 receptor mRNA level in rat cortical cells. AII (10(-9)-10(-5) M) exerted a marked increase of AT2 receptor mRNA in a dose dependent manner. The maximum increase was observed at 3 hr of AII stimulation and lasted 3 hr. The up-regulation of AT2 receptor mRNA was antagonized by PD123319, an AT2 receptor antagonist, but not by SC-52458, an AT1 receptor antagonist, thus suggesting that the increase in AT2 receptor mRNA is mediated via AT2 receptor. This increase is blocked by serine/threonine phosphatase inhibitor okadaic acid, but not by the phosphotyrosine phosphatase inhibitor sodium vanadate, thus suggesting the involvement of serine/threonine phosphatase in this process. Protein kinase C inhibitor, H-7 and calphostin C, did not inhibit the AII-induced up-regulation significantly. In addition, calcium ionophore, A23187 had no effect. These findings suggest that the AT2 receptor mRNA expression by AII is regulated by the activity of serine/threonine phosphatase in the cortical neurons. This observation is also the first example concerning the regulation of AT2 receptor within the brain. PMID- 9344884 TI - The neurotoxic effects of ochratoxin-A are reduced by protein binding but are not affected by l-phenylalanine. AB - Recent in vivo investigations indicate that the mycotoxin ochratoxin A (OTA) is a neurotoxicant during prenatal stages. In line with in vivo data, in our embryonic chick brain and neural retina cell cultures the markers for neuritic outgrowth and differentiation (NF68 and 160 kDa, MAP2 and MAP5) were especially negatively affected. In vivo OTA is nearly completely bound to serum constituents. In our culture system binding of OTA to BSA evoked a significant shift of the concentration-effect relationships in meningeal and brain cell cultures. As a result of the albumin binding the OTA IC5 and IC50 values of all parameters increased by nearly the same value (about 15-fold in brain and 32-fold in meningeal cell cultures). One of the mechanisms responsible for OTA toxicity is thought to be the competitive inhibition versus Phe of Phe-dependent enzymes. Therefore, in addition, we investigated the effects of l-phenylalanine (Phe) and its influence on OTA toxicity in brain and neural retina cell cultures. Phe itself was found to differently affect brain and neural retina cell cultures. However, in both cultures OTA toxicity is not diminished by Phe. Therefore, our data indicate that at least in our cultures competition with Phe-dependent processes does not play a role in OTA toxicity. PMID- 9344885 TI - Cytokine production by human airway epithelial cells after exposure to an air pollution particle is metal-dependent. AB - Despite the many epidemiological studies supporting the contention that ambient air pollution particles can adversely affect human health, there is no clear agreement as to a biologically plausible mechanism which can explain the acute mortality and morbidity associated with exposure to particles less than 10 microm in size. We tested the hypothesis that metals present in an air pollution particle can induce the synthesis and expression of the inflammatory cytokines IL 8, IL-6, and TNFalpha. A residual oil fly ash (ROFA) containing the transition metals vanadium, nickel, and iron was used as a model emission source air pollution particle. Normal human bronchial epithelial (NHBE) cells were exposed for either 2 or 24 hr to 0, 5, 50, or 200 microg/ml ROFA. Concentrations of IL-8, IL-6, and TNF-alpha proteins were measured with commercially available ELISA kits. mRNA for these same cytokines was quantified by RT-PCR. NHBE cells exposed to ROFA produced significant amounts of IL-8, IL-6, and TNF, as well as mRNAs coding for these cytokines. Cytokine production was inhibited by the inclusion of either the metal chelator deferoxamine (1.0 mM) or the free radical scavenger dimethylthiourea (1.0 mM). In addition, vanadium containing compounds, but not iron or nickel sulfates, mimicked the effects of intact ROFA. These results demonstrate that metals present in ROFA may be responsible for production and release of inflammatory mediators by the respiratory tract epithelium and suggest that these mediators may contribute to the toxic effects of particulate air pollutants reported in epidemiology studies. PMID- 9344886 TI - Pharmacokinetics and metabolism of dichloroacetate in the F344 rat after prior administration in drinking water. AB - The effect of prior administration of dichloroacetate (DCA) in drinking water on the pharmacokinetics of DCA in male F344 rats was studied. Rats were provided with DCA in their drinking water at 0.2 and 2.0 g/liter for 14 days and then challenged with iv bolus iv or gavage doses of [14C1,2]DCA, 16 hr after pretreatment withdrawal. The blood concentration-time profiles of DCA and the disposition of 14C was characterized and compared with controls. The effect of pretreatment on the in vitro metabolism of DCA in hepatic cytosol was also evaluated. Pretreatment caused a significant increase in the blood concentration and AUC0-->infinity of DCA (433.3 versus 2406 microg ml-1 hr). Pharmacokinetic analysis indicated that pretreatment significantly decreased total body clearance (267.4 versus 42.7 ml hr-1 kg-1), which was largely due to decreased metabolism since only modest differences in the urinary clearance of DCA were observed. Pretreatment significantly decreased the formation of 14CO2 after both iv and oral doses of [14C]DCA. The decrease in CO2 formation was also observed after pretreatment with DCA at 0.2 g/liter. Pretreatment also increased the urinary elimination of DCA and several metabolites, particularly glycolate. The in vitro experiments demonstrated that DCA pretreatment inhibited the conversion of DCA to glyoxylate, oxalate, and glycolate in hepatic cytosol. These results indicate that DCA has an auto-inhibitory effect on its metabolism and that pharmacokinetic studies using single doses in naive rats will underestimate the concentration of DCA at the target tissue during chronic or repeated exposures. PMID- 9344887 TI - Comparison of inflammatory lung responses in Wistar rats and C57 and DBA mice following acute exposure to cadmium oxide fumes. AB - Inhalation of cadmium oxide (CdO) is a significant form of human exposure to cadmium (Cd). Furthermore, there is epidemiological and experimental data relating Cd inhalation with lung cancer. Animal studies indicate that rats are more susceptible to Cd-induced lung cancer than mice, but interstrain sensitivity differences to Cd-induced pulmonary inflammation or carcinogenesis have not been addressed in either species. We compared pulmonary inflammatory processes in Wistar Furth (WF) rats with those in C57 and DBA mice exposed to freshly generated CdO fumes in nose-only inhalation chambers. Animals were exposed to 1 mg Cd/m3 for 3 hr and terminated immediately or 1, 3, and 5 days after exposure. Control animals were exposed to air/argon furnace gases. Cd-induced lung injury was assessed by bronchoalveolar lavage fluid (BALF) analyses, histopathology, and immunohistochemical detection of cell proliferation. Inhalation of CdO resulted in pulmonary inflammatory processes that varied widely across species and strains. C57 mice responded with faster and greater influx of neutrophils and proliferation of alveolar macrophages, type II epithelial cells, and bronchiolar epithelial cells compared to DBA mice or WF rats. DBA mice retained a greater percentage of inhaled Cd in the lungs and presented higher levels of BALF protein than C57 mice or rats. In comparison to mice, WF rats responded with a more transient inflammatory response in BALF parameters and higher degree of acute inflammation in lung tissue. The more pronounced proliferation of alveolar and bronchiolar epithelial cells observed in C57 mice might indicate higher susceptibility of this mice strain to Cd-induced lung carcinogenesis compared to DBA mice or WF rats. Furthermore, the present results of fewer inflammatory cells and lower proliferation of epithelial cells in DBA mice in association with our previous observation of higher Cd-induced metallothionein protein in this strain suggest that DBA might be less susceptible to the pulmonary carcinogenic effects of inhaled Cd than C57 mice or WF rats. We conclude that mice might not necessarily be more resistant than rats to the carcinogenic effects of inhaled Cd, since intraspecies susceptibility differences are strongly suggested by the present data. An extrapolation of this conclusion is that genetic variations in the human population may determine individual sensitivity differences to inhaled Cd. PMID- 9344888 TI - Subchronic/chronic toxicity of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) in rats. Part I. Design, general observations, hematology, and liver concentrations. AB - Groups of 20 male and 20 female adult Sprague-Dawley rats were given five different doses of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD). Total doses for males and females (30.9/18.5, 370/222, 2222/1333, 6667/4000, and 10000/6000 microg/kg) were divided into four daily loading doses and six biweekly maintenance doses. Positive controls were administered 2,3,7, 8 tetrachlorodibenzo-p-dioxin (TCDD, total dose 70/41.9 microg/kg). Liver concentrations, as determined by GC/MS, reflected quite accurately the calculated dose ratios. The dosing period was 13 weeks, after which half of the rats were necropsied and the rest provided with an off-dose period of another 13 weeks. Body weight gain was dose-dependently reduced throughout the study. Mortality occurred dose-dependently, starting on Day 22 and continuing until the end of the off-dose period. Mortality rates at the end of the off-dose period were 90 and 40% for males and 60 and 10% for females in the two highest dose groups. Clinical signs and necropsy findings suggested that the cause of death was related to wasting (early deaths), gastrointestinal and nasal hemorrhage (between Days 64 and 126), or anemia (late deaths, after Day 111). Prothrombin times were prolonged intermittently, mainly at the highest dose of HpCDD. Platelet counts were dose-dependently decreased at the two highest doses of HpCDD and in the TCDD treated group. This study demonstrates that the relative potency derived from acute toxicity studies is the same as that observed in this subchronic/chronic toxicity study of HpCDD and TCDD, confirming the validity of 0.007 as the toxic equivalency factor (TEF) for HpCDD, which is in good agreement with the international TEF of 0.01. PMID- 9344889 TI - Subchronic/chronic toxicity of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) in rats. Part II. Biochemical effects. AB - Groups of 20 male and 20 female rats were given five different oral doses of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) or one dose of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) divided into four daily loading doses and six biweekly maintenance doses. The dosing period was 13 weeks, after which half of the rats were necropsied and the rest assigned to an off-dose period of another 13 weeks. At the end of the dosing period, liver ethoxyresorufin O-deethylase (EROD) activity was dose-dependently increased starting at the lowest dose (7- to 10-fold) with maximum induction (50- to 100-fold) at the middle or second highest dose. There was a slight reversibility of this effect in HpCDD-treated rats, particularly at lower doses, and a pronounced reversibility in TCDD-dosed rats, both in accordance with respective toxicokinetics. The activity of phosphoenolpyruvate carboxykinase in liver was dose-dependently decreased (up to 60%) at the two or three highest doses of HpCDD and also in the TCDD dosage group. Liver tryptophan 2,3-dioxygenase activity was decreased at the two highest doses of HpCDD (up to 41%), particularly in females. Serum tryptophan concentrations were elevated in rats found moribund due to wasting. There was a dose-dependent decrease in serum glucose concentrations (up to 30%) at the end of the dosing period. Serum thyroxin (T4) concentrations showed a dose-dependent decrease (78% at the highest dose) beginning in the middle dose for HpCDD and in the TCDD dosage group. Serum triiodothyronine (T3) concentrations were only slightly affected, except that they were somewhat decreased in moribund animals. The results demonstrate that similar biochemical changes occur in rats after single as after multiple dosing with HpCDD and TCDD. Based on these endpoints, the relative potency of HpCDD after subchronic exposure is in agreement with the international toxic equivalency factor (I-TEF) of 0.01 and, more specifically, with a TEF of 0.007 based on LD50 values in the same strain of rats. PMID- 9344890 TI - Chlorpyrifos, parathion, and their oxons bind to and desensitize a nicotinic acetylcholine receptor: relevance to their toxicities. AB - The nicotinic acetylcholine receptor (nAChR) of the electric organ of the electric ray. Torpedo sp., the richest source of nAChR, with similar structure and pharmacology to the mammalian skeletal muscle nAChR, carries several binding sites for different ligands. Incubation of Torpedo membrane-bound nAChRs with the agonist carbamylcholine (Carb) stimulated the binding of [3H]thienyl cyclohexylpiperidine ([3H]TCP), which binds to the receptor's noncompetitive antagonist binding site in its ionic channel, with high affinity (Kd of 196 nM). The agonist-stimulated binding of [3H]TCP (i.e., binding to activated nAChRs) was inhibited in a concentration-dependent manner by four organophosphate (OP) anticholinesterases, chlorpyrifos oxon (CPO), chlorpyrifos (CPS), parathion (PS), and paraoxon (PO) with IC50 (concentration that inhibits 50% of the effect) values of 5, 150, 200, and 300 microM, respectively. The binding of CPO was totally reversible. The OPs had no effect on equilibrium binding of [alpha 125I]bungarotoxin ([alpha-125I]BGT) to the receptor's acetylcholine (ACh)-binding site, but preincubation of the membranes with the OPs increased this site's affinity for Carb. In absence of agonist, 100 microM of the OPs increased the binding of [3H]TCP by two- to fivefold with the following order of decreasing potency: PS > CPO > CPS > PO. The data suggest that in addition to inhibition of acetylcholinesterase, these OPs bind to a site on the nAChR that is different from the sites that bind ACh or TCP and that this binding induces nAChR desensitization. The relevance of this direct action of OPs on nAChRs on their acute toxicities is discussed. PMID- 9344891 TI - In utero exposure to low doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin alters reproductive development of female Long Evans hooded rat offspring. AB - Prenatal administration of a single dose of 1 microg TCDD/kg induces malformations of the external genitalia and subfertility in female offspring (L. E. Gray, Jr., and J. S. Ostby (1995) Toxicol. Appl. Pharmacol. 133, 285-294). A cross-fostering study indicated that in utero but not lactational TCDD exposure (1 microg TCDD/kg on gestational Day 15) induces cleft phallus, vaginal thread formation, and reduced ovarian weight. TCDD treatment on the 15th day of pregnancy at 0, 0.05, 0.20, or 0.80 microg TCDD/kg delayed vaginal opening at 0.80 microg/kg in the progeny. A persistent vaginal thread was displayed by 27% of the progeny at 0.20 and 92% at 0.80 microg TCDD/kg. These effects did not appear to result from abnormal ovarian function during prepubertal development; neither serum estradiol levels nor ovarian estradiol production were reduced in 21- or 28-day-old progeny of dams exposed to 1 microg TCDD/kg. In addition, partial to complete clefting of the phallus was displayed in TCDD-treated rats (10% at 0.20 and 60% at 0.80 microg TCDD/kg) and these dosage levels also increased the length of the urethral slit, increased distance from the urethral opening to the tip of the phallus, and decreased distance from the urethral opening to the vaginal orifice. Although fertility rates were normal, time-to pregnancy was delayed by treatment with 0.80 microg TCDD/kg. When necropsied at 20 months of age, females from the TCDD-dose groups displayed histopathological alterations of the reproductive tract. In summary, administration of TCDD at dosage levels of 0.2, 0.8, and 1.0 microg/kg produces morphological reproductive alterations in female rat offspring as a consequence of in utero exposure. PMID- 9344892 TI - Metabolism of ethyl carbamate by pulmonary cytochrome P450 and carboxylesterase isozymes: involvement of CYP2E1 and hydrolase A. AB - The lung is highly susceptible to ethyl carbamate (EC)-induced tumorigenesis. Our goal in this study was to investigate the in vitro isozyme-selective metabolism of EC in lung microsomes by cytochrome P450 and carboxylesterase enzymes. Our results showed that incubations with EC produced significant reduction in p nitrophenol (PNP) hydroxylation and N-nitrosodimethylamine (NDMA) demethylation; there were no alterations in 7-pentoxyresorufin- and 7-ethoxyresorufin O dealkylase activities. Reaction of microsomes with an inhibitory CYP2E1 antibody and subsequent reaction with EC abolished the EC-induced diminution in NDMA demethylase activity. Carboxylesterase activity, as assessed by hydrolysis of p nitrophenyl acetate, was significantly decreased in microsomes incubated with EC. Reactions with EC in conjunction with the carboxylesterase inhibitors, paraoxon (PAX) or phenylmethylsulfonyl fluoride (PMSF), abolished the EC-induced decrease in carboxylesterase activity; PAX is a broad-spectrum carboxylesterase inhibitor, whereas PMSF is a specific inhibitor of hydrolase A, a carboxylesterase isozyme. Incubations of EC in combination with either PAX or PMSF exacerbated the EC induced reduction in PNP hydroxylase and NDMA demethylase activities. Alterations in immunodetectable CYP2E1 protein levels were not apparent in microsomes incubated with EC alone, but the amounts were decreased in reactions with EC in conjunction with either PAX or PMSF. Immunoblotting with antibodies for the carboxylesterase isozymes, hydrolase A and B, revealed loss of immunodetectable hydrolase A in microsomes incubated with EC, PAX, or PMSF. However, immunodetectable hydrolase B was only decreased in microsomes reacted with PAX but not with PMSF or EC. These findings correlated with our covalent binding data, which showed that levels of binding of [14C-ethyl]EC to lung microsomes were significantly higher in incubations conducted in conjunction with PAX or PMSF, compared with control levels. Antibody inhibition of the CYP2E1 enzyme significantly reduced the extent of binding. Our results demonstrated that EC metabolism in lung microsomes, as estimated from magnitudes of covalent binding, is mediated by the P450 isozyme CYP2E1 and the carboxylesterase isozyme hydrolase A. PMID- 9344893 TI - Do endogenous volatile organic chemicals measured in breath reflect and maintain CYP2E1 levels in vivo? AB - The effect of trans-1,2-dichloroethylene (DCE), an inhibitor of cytochrome P450 (P450) 2E1 (CYP2E1), on the composition and quantity of volatile organic chemicals (VOCs) expired in the breath of male F-344 rats was determined in parallel with hepatic P450 activity and content. Hepatic microsomes were prepared from groups of rats prior to dosing and at 2, 5, 12, and 24 hr postdosing with DCE (100 mg/kg ip), and total P450 content and the activity of CYP2E1 was determined. Breath was collected from parallel groups of rats predose and at several intervals that encompassed the time points for rats euthanized for microsome preparation. Over 100 breath components were identified by GC/MS and quantitated by GC/FID. The overall change in the profile of breath VOCs resulting from administration of DCE was striking. An increase of approximately 1000% was measured in the mass of non-DCE-derived VOCs exhaled 4-6 hr after dosing, but there was no increase in hepatic lipid peroxidation. In addition to hexane, short chain methyl ketones were particularly affected, and percentage increases in response to inhibition were inversely related to chain length, with acetone and 2 butanone > 2-pentanone >> 2-hexanone > 2-heptanone. There were no statistically significant decreases in total content of P450, but the activity of CYP2E1 was diminished about 65% at 2 and 5 hr after DCE treatment. However, 24 hr after inhibitor administration the total mass of VOCs expired was only marginally elevated above baseline and CYP2E1 activity was not significantly different from that of untreated rats. The compounds most markedly increased upon inhibition of CYP2E1 are also excellent inducers of that isozyme, and this finding is consistent with the hypothesis that these chemicals are important to the normal homeostasis of CYP2E1. The increase in breath components observed following inhibition of CYP2E1 suggests that VOCs in breath can reflect the activity of that isozyme in vivo. PMID- 9344894 TI - Effects of antioxidants and Ca2+ in cisplatin-induced cell injury in rabbit renal cortical slices. AB - Effects of antioxidants, reactive oxygen species (ROS) scavengers, and Ca2+ on cisplatin-induced renal cell injury were studied in rabbit renal cortical slices in vitro. Cisplatin induced LDH release and lipid peroxidation, inhibition of PAH uptake, and GSH depletion. These changes were significantly prevented by thiols (DTT and GSH), antioxidants (DPPD and BHA), and an iron chelator (deferoxamine). Superoxide dismutase partially reduced the cisplatin-induced LDH release without affecting the lipid peroxidation and the GSH depletion. Catalase did not affect the LDH release and the lipid peroxidation induced by cisplatin. Hydroxyl radical scavengers prevented the lipid peroxidation, whereas they did not alter the LDH release, the inhibition of PAH uptake, and the GSH depletion induced by cisplatin. Removal of Ca2+ or addition of EGTA to the incubation medium did not alter cisplatin effects on LDH release and lipid peroxidation. Buffering intracellular Ca2+ with quin-2/AM or inhibition of intracellular Ca2+ release with TMB-8 significantly reduced the cisplatin effect on LDH release without any effect on the lipid peroxidation and the GSH depletion. Ruthenium red attenuated the LDH release, the lipid peroxidation, and the inhibition of PAH uptake mediated by cisplatin. La3+ prevented the cisplatin effect on the LDH release, whereas it did not affect the lipid peroxidation, the inhibition of PAH uptake, and the GSH depletion by cisplatin. These results suggest that cisplatin induces a lethal cell injury by lipid peroxidation-dependent and -independent mechanisms and that the cell injury and the lipid peroxidation by cisplatin are iron dependent. In addition, the data indicate that the Ca2+ released from intracellular stores, but not the Ca2+ moved from extracellular space, plays a role in the cisplatin-induced cell injury independent of lipid peroxidation. PMID- 9344895 TI - Induction of apoptotic cell death by particulate lead chromate: differential effects of vitamins C and E on genotoxicity and survival. AB - Certain hexavalent chromium compounds are documented human carcinogens. Exposure of cells to particulate forms of chromium results in cell-enhanced dissolution of particles in the extracellular microenvironment and chronic production of chromium oxyanions, which are taken up by the cell through an anion transport system and are genotoxic and clastogenic. It was previously shown that apoptosis is the mode of cell death of nearly all of the Chinese hamster ovary cells (CHO AA8 cell line), which die after high-dose, short-term treatments with soluble sodium chromate. In this report the mode of cell killing by particulate lead chromate and of low-dose continuous treaments of soluble sodium chromate designed to mimic conditions of ionic chromate uptake after lead chromate exposure was examined. CHO-AA8 cells were treated for 24 hr with doses of sodium chromate or lead chromate which cause a 50% decrease in survival in colony-forming effeciency assays. Longer treatments (up to 72 hr) at the same doses did not decrease survival further than the 24-hr exposure. Morphological changes indicative of apoptosis, as well as internucleosomal DNA fragmentation, were detectable by 24 hr after treatment with lead chromate or soluble sodium chromate. All of the cells killed by treatments with lead chromate particles underwent apoptosis as the mode of cell death and this was accurately modeled in cell culture by continuous treatments with low-dose soluble sodium chromate. Exposure of cells to hexavalent chromium compounds causes a spectrum of DNA damage which can be selectively altered by pretreatment of cells with antioxidant vitamins prior to chromium exposure. Here we show that ascorbate and alpha-tocopherol markedly inhibited the chromosomal aberrations induced by both particulate and soluble chromate compounds, even though chromium adduct levels were not decreased by either vitamin pretreatment. Cell survival assays showed that ascorbate, but not alpha-tocopherol, protected cells from apoptosis induced by sodium chromate. The results differentiate chromium-induced apoptosis from both chromosomal damage and adduct levels and suggest that other lesions sensitive to ascorbate but not tocopherol are the proximal inducing signal for chromium-induced apoptosis. PMID- 9344896 TI - Hepatic foci in rats after diethylnitrosamine initiation and 2,3,7,8 tetrachlorodibenzo-p-dioxin promotion: evaluation of a quantitative two-cell model and of CYP 1A1/1A2 as a dosimeter. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent hepatic tumor promoter in female rats. We used a quantitative, stochastic initiation-promotion model based on R. B. Conolly and J. S. Kimbell (Toxicol. Appl. Pharmacol. 124, 284-295, 1994) to analyze initiation-promotion results from a previously published study (H. C. Pitot et al., Carcinogenesis 8, 1491-1499, 1987) within the context of a negative selection model of tumor promotion. In this model, two types of initiated cells (called A and B cells) are produced by DEN initiation. Visually excellent correspondence between model predictions and data (i.e., foci/cm3 liver and percentage of liver occupied by foci) are obtained when TCDD is described as having dose-responsive effects on division and death (apoptotic) rates of these two cell types. For A cells, both the division and the death rates increase while the difference between division and apoptotic rates decreases. For B cells, the difference between division and apoptotic rates increases, primarily due to a decrease in the apoptotic rate. We also linked these alterations in cell kinetics to a pharmacokinetic model for TCDD incorporating a five subcompartment model of the liver acinus with induction of CYP1A1 and 1A2 genes in the subcompartments. Alterations in A cell kinetics correlate with effects of TCDD in the region most sensitive to induction (subcompartment 5-centrilobular region); B cell dynamics correlate with induction in subcompartments 3-5 (centrilobular and mid-zonal regions). In summary, these modeling exercises show that (1) the two-cell model, without presuming effects of TCDD on the mutation rate of normal hepatocytes, reproduces the data of Pitot et al. (1987) and (2) induction of CYP1A1/1A2 in different regions of the hepatic acinus can be used as a general correlate of these presumed changes in cell growth kinetics. PMID- 9344897 TI - Mercury uptake by LLC-PK1 cells: dependence on temperature and membrane potential. AB - The purpose of this study was to investigate the mechanism of inorganic mercury (Hg) uptake in LLC-PK1 cells, a renal tubular epithelial cell line, and to compare the results with those reported previously by us in rat renal cortical epithelial (RCE) cells in primary culture. The LLC-PK1 cells were cultured for 3 12 days, incubated with 1 microM HgCl2 in Hanks' balanced salt solution at 4 or 37 degrees C for 30 min, and washed with phosphate-buffered saline containing BAL to remove the cell membrane-bound Hg. The uptake of Hg was higher in nonconfluent cultures than in confluent cultures and higher at 37 than at 4 degrees C. In confluent culture (Day 8) Hg uptake at 4 degrees C was only 27% of that at 37 degrees C. The initial accumulation of Hg (5 min) from different concentrations of HgCl2 (0.5-50 microM) was linear and did not show a tendency toward saturation, suggesting that a carrier-mediated process was not involved. Pretreatment of cells with 10 microM FCCP, a metabolic inhibitor and a proton ionophore, 0.5 mm DIDS, an anion transport inhibitor, or 0.5 mM ouabain, a Na+/K+ ATPase inhibitor, resulted in 72, 60, and 57% reduction in Hg uptake, respectively. Furthermore, replacement of 137 mm NaCl in the incubation medium with 137 mM KCl or LiCl or 274 mM mannitol caused 30, 45, and 87% reduction in Hg uptake, respectively. These results suggest that in LLC-PK1 cells, as in RCE cells, Hg uptake is inversely related to cell density and is influenced by membrane fluidity, membrane potential, and HCO3-/Cl- transporter. PMID- 9344898 TI - Structure-activity relationship for two lipoxygenase inhibitors and their potential for inducing nephrotic syndrome. AB - In a study of structure-activity relationship with drug-induced nephropathy two lipoxygenase inhibitors, the N-hydroxyurea derivative 70C ((E)-N-{3-[3-(4 fluorophenoxy) phenyl]-1-(R, S)-methylprop-2-enyl}-N-hydroxyurea) and the N hydroxamic acid analogue 360C ((E)-N-{3-[3-(4-fluorophenoxy) phenyl]-1-(R, S) methylprop-2-enyl}-N-hydroxamic acid), were administered to rats. 70C and 360C were dosed to female Wistar rats at 100 mg/kg po daily for 7 days. Another group of rats was given a single intravenous bolus dose of puromycin aminonucleoside (PAN) at 100 mg/kg. Urine samples were collected from all groups during the study and plasma samples were collected after 7 days. Kidneys were excised and fixed for examination by electron microscopy. 70C- and PAN-treated groups both showed early changes in the glomeruli, in which the visceral cells appeared enlarged and showed varying degrees of foot process loss. This foot process loss was associated with decreases in total plasma protein and albumin and increases in the plasma cholesterol, triglycerides, creatinine, and urea were recorded. Marked proteinuria was observed in both the 70C and PAN groups. The foot process loss together with increased proteinuria, hypoalbuminemia, hypercholesterolemia, and lipemia are all characteristic of the human condition, Minimal Change Nephrotic Syndrome. All the biochemical and morphological investigations showed that 360C treated rats were similar to the control group, suggesting that the hydroxyurea moiety of 70C is responsible, either directly or indirectly, for the induction of the nephrotic syndrome seen in rats. PMID- 9344899 TI - Acute lung failure induced by tricyclic antidepressants. AB - Overdosing of several drugs, such as tricyclic antidepressants, salicylates, and opiates, is known to induce effects like those seen in patients with adult respiratory distress syndrome. By exposing isolated perfused and ventilated rat lungs via the perfusate to six different tricyclic antidepressants (amitriptyline, nortriptyline, imipramine, desipramine, mianserine, and maprotiline), we investigated possible effects on ventilation (conductance and dynamic compliance), lung perfusion flow, and edema formation. The effects of these substances were pronounced and appeared within 15 min after exposure. Amitriptyline was studied in greater detail and caused a dose-related (0.01-1.0 mM) reduction in ventilation and perfusion flow. At the highest drug concentration pronounced lung edema was observed. Morphological studies were conducted with a transmission electron microscope. The microscopic preparations showed dose-related edema (amitriptyline 0.1 and 1.0 mM). The effects noted in our experimental studies are similar to those described in patients who have taken an overdose of tricyclic antidepressants. This emphasizes the possibility of a noncardiogenic edema component in these patients. PMID- 9344901 TI - LETTERS TO THE EDITOR PMID- 9344902 TI - Paramyxovirus fusion (F) protein and hemagglutinin-neuraminidase (HN) protein interactions: intracellular retention of F and HN does not affect transport of the homotypic HN or F protein. AB - To investigate a possible intracellular coassociation of the paramyxovirus simian virus 5 (SV5) and human parainfluenza virus type 3 (HPIV-3) fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins in a living cell, without resorting to chemical crosslinking and antibody coimmunoprecipitation, we tagged the cytoplasmic N-terminus of SV5 HN with a RRRRR motif and HPIV-3 HN with a RRR motif for endoplasmic reticulum (ER) retention. In addition, we tagged the cytoplasmic C-terminus of SV5 and HPIV-3 F with a KK motif. The RRR- or RRRRR tagged HN molecules were coexpressed in mammalian cells together with the homologous wt F proteins, and the KK-tagged F molecules were coexpressed with the homologous wt HN proteins, and in each case the transport of the wt F or HN molecules was investigated. The data suggest that an association of F and HN of sufficient affinity to alter the transport of the reporter molecule does not occur intracellularly in the ER or the Golgi apparatus. PMID- 9344900 TI - Inhibition of carbamyl phosphate synthetase-I and glutamine synthetase by hepatotoxic doses of acetaminophen in mice. AB - The primary mechanisms proposed for acetaminophen-induced hepatic necrosis should deplete protein thiols, either by covalent binding and thioether formation or by oxidative reactions such as S-thiolations. However, in previous studies we did not detect significant losses of protein thiol contents in response to administration of hepatotoxic doses of acetaminophen in vivo. In the present study we employed derivatization with the thiol-specific agent monobromobimane and separation of proteins by SDS-PAGE to investigate the possible loss of specific protein thiols during the course of acetaminophen-induced hepatic necrosis. Fasted adult male mice were given acetaminophen, and protein thiol status was examined subsequently in subcellular fractions isolated by differential centrifugation. No decreases in protein thiol contents were indicated, with the exception of a marked decrease in the fluorescent intensity, but not of protein content, as indicated by staining with Coomassie blue, of a single band of approximately 130 kDa in the mitochondrial fractions of acetaminophen-treated mice. This protein was identified by isolation and N terminal sequence analysis as carbamyl phosphate synthetase-I (CPS-I) (EC 6.3.4.16). Hepatic CPS-I activities were decreased in mice given hepatotoxic doses of acetaminophen. In addition, hepatic glutamine synthetase activities were lower, and plasma ammonia levels were elevated in mice given hepatotoxic doses of acetaminophen. The observed hyperammonemia may contribute to the adverse effects of toxic doses of acetaminophen, and elucidation of the specific mechanisms responsible for the hyperammonemia may prove to be useful clinically. However, the preferential depletion of protein thiol content of a mitochondrial protein by chemically reactive metabolites generated in the endoplasmic reticulum presents a challenging and potentially informative mechanistic question. PMID- 9344903 TI - Activation of the envelope proteins by a metalloproteinase enables attachment and entry of the hepatitis B virus into T-lymphocyte. AB - Previously, we identified an HBV binding factor (HBV-BF), a 50-kDa serum glycoprotein which interacts with HBV envelope proteins and which is also located in the membrane of normal human hepatocyte (A. Budkowska et al. (1993) J. Virol. 67, 4316). Here we show that HBV-BF is a neutral metalloproteinase which shares substrate specificity and properties with a newly described family of membrane type matrix metalloproteinases. HBV-BF treatment of the HBV resulted in the cleavage of the N-terminal part of the middle HBV envelope protein at the pre S2(136-141) amino acid sequence VRGLYF/L (containing a single arginine cleavage site). HBV-BF affected the reactivity of the large HBV protein with pre-S1 specific MAbs, probably inducing the conformational change of the pre-S1 domain. The HBV-BF-digested virus remained morphologically intact with unchanged S antigenic determinants. The structural modifications of the viral envelope proteins induced by HBV-BF enabled cell membrane attachment and viral entry into the T-lymphocyte. Both processes were blocked by the metalloproteinase inhibitor 1,10 phenanthroline. Thus, the host-dependent proteolytic activation of the envelope proteins seems to be essential for the HBV entry into the cell. HBV-BF under a membrane bound or a secreted form could be (one of) the molecule(s) responsible for the HBV proteolytic activation. PMID- 9344904 TI - Differential effects on HIV-1 gene regulation by EBV in T lymphocytic and promonocytic cells transduced to express recombinant human CR2. AB - A panel of human hematopoietic cell lines was genetically engineered to express recombinant complement receptor 2 (CR2 or CD21), which is also the Epstein-Barr virus (EBV) receptor. The panel was composed of SupT1, J1.1, and U1.HIV cells. The latter is a promonocytic cell line, whereas the other two are T lymphocytic cell lines. J1.1 and U1.HIV cells are latently infected by human immunodeficiency virus type 1 (HIV-1). These three cell lines were transduced with a murine leukemia virus (MLV)-based retroviral vector system. CR2 was efficiently and consistently expressed on the cell membranes, conferring enhanced susceptibility to EBV infection. The efficient expression of recombinant CR2 in cell lines of hematopoietic origin allowed for study of the interaction between EBV infection and HIV-1 gene regulation in suitable cell-culture models. The effects of EBV and HIV-1 coinfection results were cell-type dependent. In the two T lymphocytic cell lines, HIV-1 expression was rapidly and persistently down-regulated by EBV. Conversely, in the promonocytic cell line U1.HIV-CR2, HIV-1 expression was transiently enhanced by EBV. The EBV and HIV-1 coinfection result in U1.HIV-CR2 cells is potentially important, as the activation of HIV-1 gene expression in monocyte-like cells may play a crucial role in the mechanism of CD4+ T cell depletion by apoptosis. Therefore, the U1.HIV-CR2 cell line may represent a useful cell-culture system to study the synergism between EBV and HIV-1 in inducing apoptosis in primary CD4+ T cells. PMID- 9344905 TI - HsN3 proteasomal subunit as a target for human immunodeficiency virus type 1 Nef protein. AB - HIV-1 Nef protein is important for pathogenicity, but its biochemical function remains obscure. To clarify its role, a yeast two-hybrid system (ths) screening was utilized to identify Nef cellular partners. Of 79 yeast clones harboring cDNAs for putative Nef binding proteins, 27 (34%) contained the coding region for HsN3 proteasomal subunit. HsN3 behaved as bona fide Nef partner in ths control crosses. Nef-HsN3 interaction was confirmed by in vitro binding experiments. In particular, recombinant Nef was able to capture the HsN3 subunit from a natural proteasome preparation. In Nef, the interacting region was mapped within aa 34 143, which span the structured portion of the protein, including the SH3-binding domain. In HsN3, Nef-binding portion was restricted to aa 73-249, and the tract 219-249-reminiscent of SH3 domain N-terminal 3/5ths-was shown to be essential, though not sufficient. Attempts to purify a Nef-HsN3 complex from transfected COS7 cells were unsuccessful. However, Nef was found to markedly downregulate intracellular levels of both a coexpressed HsN3 and the endogenous simian homologue. These results suggest that Nef, by binding to a subunit, might alter proteasome function in infected cells. PMID- 9344906 TI - The HIV-1 matrix domain of Gag is required for Vpu responsiveness during particle release. AB - HIV-1 viral protein U (Vpu) facilitates virus particle release. To determine whether Gag is sufficient for generation of a target for Vpu-mediated particle release, we expressed HIV-1 Gag protein in the absence of the other viral genes. The resulting particles were still Vpu responsive. Mutational analysis of Gag indicated that the matrix domain (MA) is required for Vpu responsiveness. However, additional mutations in other domains of Gag, which affect the formation of stable virus particles, also abrogate Vpu responsiveness on total Gag release. Coexpression of the wild-type gag gene and a gag mutant lacking the MA domain renders the MA- mutant Vpu responsive. This indicates that Gag molecules lacking MA are still incorporated into particles through association with wild-type Gag molecules and that the resulting composite particles are sufficient for Vpu mediated exit. PMID- 9344907 TI - The enigma of pX: A host-dependent cis-acting element with variable effects on tombusvirus RNA accumulation. AB - Tomato bushy stunt virus (TBSV) is a small isometric virus that contains a single stranded RNA genome with five major genes. In this study, we have analyzed the importance of an additional small sixth open reading frame (ORF) of 207 nucleotides, designated pX, which resides at the 3' end of the genome. Bioassays showed that deletions or additions of nucleotides at the 5' end of the pX gene that were designed to disrupt the ORF, or site-specific inactivation of its start codon, all gave rise to TBSV mutants which were unable to accumulate to detectable levels in cucumber or Nicotiana benthamiana protoplasts. Although these results suggested a role for the putative pX protein, introduction of a premature stop codon in the pX gene had no strong negative effect. However, a comparable mutation that affected the same nucleotides without changing the predicted amino acid sequence greatly reduced RNA accumulation. Therefore, we hypothesize that cis-acting RNA sequences within the pX gene, rather than the predicted protein influence genome accumulation. The requirement of the cis acting pX ORF sequences appears to be host-dependent because comparisons revealed that subtle pX gene mutations that prohibited accumulation of TBSV RNA in cucumber or N. benthamiana, failed to interfere substantially with replication in Chenopodium quinoa protoplasts or plants. Irrespective of the host, the cis acting pX gene sequences were dispensable on replicase-deficient RNAs that require helper TBSV for replication in trans. In addition, the pX gene was not essential for in vitro translation of replicase proteins from genomic RNA. These results suggest that neither translation nor polymerase activity of the replicase proteins require pX gene sequences. However, it is possible that very early in the replication cycle of genomic RNA in vivo, the pX gene cis-acting element is essential for some other unidentified function which involves interaction with one or more host components whose composition varies slightly between different plants. PMID- 9344908 TI - Induction of intracellular membrane rearrangements by HAV proteins 2C and 2BC. AB - Hepatitis A virus (HAV) is distinguished from other picornaviruses by its slow and relatively poor, noncytopathic growth in cultures of mammalian cells. The 2C and 2BC proteins of HAV have been implicated in the determination of virus growth in cultured cells. The homologous proteins from other picornaviruses, such as poliovirus, have been demonstrated to exhibit multiple activities, such as RNA binding, nucleotide binding and NTPase, and membrane binding and reorganization. At least some of these activities are required for viral RNA replication. We report here that HAV 2C and 2BC proteins, like their poliovirus counterparts, can induce rearrangement of intracellular membranes and directly or indirectly interact with membranes. Therefore, the inefficient replication properties of HAV are not consequences of the inherent ability of 2C (2BC) to interact with membranes. The effect of 2C (2BC) protein sequences derived from a cell culture adapted (cc) strain of HAV was compared with that of corresponding protein sequences from either a wild-type (wt) strain of HAV or a faster replicating cytopathic (cp) strain. The analysis demonstrated that mutations acquired in wt virus during adaptation to cell culture do not change dramatically either the ability of these proteins to associate with membranes and induce membrane alterations or the specific architecture of the induced membrane structures. On the other hand, 2C, but not 2BC, protein from the cp strain of HAV induced different membrane structures. PMID- 9344909 TI - Determinants of substrate specificity in the NS3 serine proteinase of the hepatitis C virus. AB - Processing of the nonstructural polyprotein of the hepatitis C virus (HCV) requires the serine-type proteinase located in the amino-terminal domain of NS3. To identify residues within NS3 determining substrate specificity, a mutation analysis was performed. Using sequence alignments and three-dimensional structure predictions, amino acids assumed to be important for specificity were replaced and the enzymes were tested in an intracellular trans-processing assay for their effects on cleavage of an NS4B-5B substrate. For some of the substitutions at positions 133, 134, 135, 136, 138, 152, 155, 157, and 169, slightly reduced processing efficiencies were observed but in no case was the substrate specificity altered. In contrast, substitutions of the phenylalanine at position 154 resulted in a modified cleavage pattern, suggesting an important role for this residue in substrate specificity. To substantiate this assumption, a panel of NS4B-5B substrates carrying different P1 residues at the NS4B/5A site were tested for cleavage by these altered proteinases. We found that substitution of Phe-154 by alanine, by valine, and particularly by threonine generated enzymes with the following affinities for aliphatic P1 residues: C > L > I > V for 154 F -> A, C = L > I > V for 154 F --> V and L > C > I > V for 154 F --> T. Neither leucine nor isoleucine nor valine was accepted by the parental NS3 proteinase, showing that Phe-154 is an important determinant for substrate specificity. Furthermore, we present evidence that Ala-157 plays an additional but minor role for this property. PMID- 9344910 TI - Comparison of the rotavirus gene 6 from different species by sequence analysis and localization of subgroup-specific epitopes using site-directed mutagenesis. AB - The nucleotide sequence of gene 6 encoding the rotavirus major capsid protein VP6 of EDIM strain (EW) was determined and compared to that of 20 previously reported strains with known subgroup specificities. Multiple alignments of amino acid sequences exhibited a high level of sequence conservation (87 to 99.2%). Site specific mutagenesis experiments were undertaken to localize regions involved in subgroup specificity. Amino acid positions 305, 315, and a region 296-299 (or 301 for equine strain H-2) were identified as contributing to subgroup epitopes. A single amino acid mutation at position 305 or 315 was sufficient to change the subgroup specificity of EW VP6 protein from non I/II to subgroup I- or subgroup II-like, respectively. Mutation at these sites may be another important mechanism for subgroup variation, along with gene reassortment. PMID- 9344911 TI - Deletion mutants of the herpes simplex virus type 1 UL8 protein: effect on DNA synthesis and ability to interact with and influence the intracellular localization of the UL5 and UL52 proteins. AB - The herpes simplex virus type 1 (HSV-1) helicase-primase, an essential component of the viral DNA replication machinery, is a trimeric complex of the virus-coded UL5, UL8, and UL52 proteins. An assembly of the UL5 and UL52 subunits retains both enzymic activities, and the UL8 protein has been implicated in modulating these functions, facilitating efficient nuclear uptake of the complex and interacting with other viral DNA replication proteins. To further our understanding of UL8, we have constructed plasmids expressing mutant proteins, truncated at their N- or C-termini or lacking amino acids internally, under the control of the human cytomegalovirus major immediate-early promoter. Deletion of 23 amino acids from the N-terminus or 33 from the C-terminus abolished the ability of UL8 to support DNA replication in transient transfection assays. None of the UL8 mutants tested exhibited a strong dominant negative phenotype in the presence of the wild-type product, although some inhibition of replication was observed with mutants lacking 165 N-terminal or 497 C-terminal amino acids. The ability of the UL8 mutants to facilitate efficient nuclear localization of UL52 in the presence of coexpressed UL5 was examined by immunofluorescence. Selected mutants were also expressed by recombinant baculoviruses and tested for interaction with UL5 and UL52 in immunoprecipitation assays. The replicative ability of the mutants was found to correlate with their ability to localize UL52 to the nucleus, but not their interaction with UL5 and UL52. This property precluded the identification of any region of UL8 important for its presumed nuclear functions. PMID- 9344913 TI - Intracellular processing of pseudorabies virus glycoprotein M (gM): gM of strain Bartha lacks N-glycosylation. AB - Genes encoding homologs of the herpes simplex virus type 1 UL10 product, glycoprotein M, are conserved in all herpesviruses investigated so far. Recently, we identified pseudorabies virus (PrV) gM as a 45-kDa structural component of purified virions. A gM-PrV mutant could be propagated in cell culture, albeit at lower titers and with delayed penetration kinetics. Thus, gM has a nonessential but modulatory function in PrV infection. PrV gM is modified by addition of an N linked glycan at a consensus sequence located between the predicted first and second hydrophobic region of the protein. This N-glycosylation site is conserved in all gM homologs sequenced so far, indicating an important functional role. To analyze intracellular processing of PrV gM, Western blot analyses were performed. In PrV-infected cells, mature 45-kDa gM as well as 33- and 35-kDa precursor forms were detectable. Presumably dimeric 90- and 70-kDa proteins were also observed. The 33- and 35-kDa proteins represent nonglycosylated and glycosylated precursors as shown by endoglycosidase digestions. Investigation of several PrV strains revealed that the UL10 product of PrV strain Bartha, an attenuated virus used as vaccine, was not modified by N-glycosylation. Sequence analysis showed that the N glycosylation consensus sequence was altered from NDT to NDA, which resulted in loss of the N-glycosylation signal. To our knowledge, this is the only gM homolog identified so far which is not N-glycosylated. To investigate whether this form of the protein is functionally competent, the UL10 gene of strain Bartha was inserted into PrV strain Kaplan by substitution of the wild-type UL10 gene. The resulting recombinant expressed a UL10 protein lacking N-glycans. In vitro replication analyses did not reveal any difference in virus production, but plaque size and penetration kinetics were slightly reduced. In summary, we show that wild-type gM is modified by N-glycosylation at one conserved site. However, although this site is highly conserved throughout the herpesviruses, loss of N glycans due to mutation of the consensus sequence had only a minor effect on propagation of PrV in cell culture. PMID- 9344914 TI - X-I and X-II open reading frames of HTLV-I are not required for virus replication or for immortalization of primary T-cells in vitro. AB - In contrast to other retroviruses of the oncovirinae subgroup, the primate and bovine leukemia viruses (HTLV, STLV, and BLV) encode genes in the X-region of the genome, between the env gene and the 3' long terminal repeat. In HTLV-I, two overlapping open reading frames (ORFs) in the distal half of the X-region encode tax and rex genes, while two ORFs (X-I and X-II) in the proximal half of this region potentially encode proteins designated p12(XI) (or rof) and p30(XII) (or tof). The biological functions and mechanisms of tax and rex have been studied extensively whereas the roles of the other ORFs have not yet been established. To identify possible functions for ORFs X-I and X-II, an infectious molecular clone of HTLV-I and a mutant provirus lacking these ORFs were compared with respect to virus replication, gene expression, and ability to immortalize primary T-cells. When transiently transfected into 293 cells, both intact and deleted proviruses directed the synthesis of virus mRNAs and proteins that were quantitatively and qualitatively identical. These viruses were also indistinguishable in their abilities to infect and replicate in DBS-FRhL cells, which are permissive for HTLV-I propagation. Immortalized T-cell lines were established after cell-free or coculture methods for infection of activated, human peripheral blood or cord blood lymphocytes with each of the cloned viruses. The growth kinetics, cytokine dependence, and cell surface markers of the infected T-cell cultures were similar for each provirus clone. Thus, ORFs X-I and X-II are not essential for virus infectivity, replication, gene expression, or T-cell immortalization in vitro. PMID- 9344912 TI - Transcriptional analysis of walleye dermal sarcoma virus (WDSV). AB - Walleye dermal sarcoma virus (WDSV) is a complex retrovirus associated with dermal sarcomas of walleye that develop and regress on a seasonal basis. WDSV contains, in addition to gag, pol, and env, three open reading frames (ORFs) designated ORF A, ORF B, and ORF C. The polymerase chain reaction technique was used to amplify and clone cDNAs representing subgenomic viral mRNAs isolated from developing (fall) and regressing (spring) tumors. Nine different singly or multiply spliced viral transcripts were identified and all were found to utilize a common 5' leader sequence. This leader sequence is spliced to the pol/env junction or downstream of env to generate singly spliced transcripts. Multiply spliced transcripts contain the 5' leader, the pol/env junction, and sequences derived from the 3' end of the genome. One multiply spliced transcript was isolated with the potential to encode the full-length ORF A protein. In addition, WDSV produced mRNAs that utilize alternative splice acceptor sites which would allow synthesis of five variant forms of the ORF A protein. In contrast, the ORF B protein is postulated to arise from a singly spliced transcript with the potential to encode the entire open reading frame. Spliced subgenomic transcripts representing ORF C mRNAs were not identified, suggesting that ORF C may be encoded from the full-length viral genomic transcript. We estimate that at least a 100-fold lower amount of the accessory/regulatory subgenomic transcripts exists in developing vs regressing tumors. These results demonstrate that WDSV undergoes an elaborate pattern of mRNA splicing similar to that of other complex retroviruses. PMID- 9344915 TI - Intact eukaryotic initiation factor 4G is required for hepatitis A virus internal initiation of translation. AB - The requirements for optimal activity of the hepatitis A virus (HAV) internal ribosome entry segment (IRES) differ substantially from those of other picornavirus IRESes. One such difference is that, to date, the HAV IRES is the only one whose efficiency is severely inhibited in the presence of the picornaviral 2A proteinase. Here we describe experiments designed to dissect the mechanism of proteinase-mediated inhibition of HAV translation. Using dicistronic mRNAs translated in vitro, we show that the HAV IRES is inhibited by the foot-and mouth disease virus Lb proteinase, as well as by the human rhinovirus 2A proteinase. Furthermore, using mutant Lb proteinase, we demonstrate that proteolytic activity is required for inhibition of HAV IRES activity. Translation inhibition correlated closely with the extent of cleavage of the one identified common cellular target for the 2A and Lb proteinases, eukaryotic initiation factor (eIF) 4G, a component of the eIF4F cap-binding protein complex. Total rescue of HAV IRES activity was possible if purified eIF4F was added to translation extracts. In contrast, if the added eIF4F contained cleaved eIF4G, no rescue of HAV IRES activity was evidenced. Thus the HAV IRES requires intact eIF4G for activity. This is unique among the picornavirus IRESes studied to date and may help explain why HAV does not inhibit host cell translation during viral infection. PMID- 9344916 TI - The locus of Epstein-Barr virus terminal repeat processing is bound with enhanced affinity by Sp1 and Sp3. AB - EBV DNA contains G-rich, repeat regions that are involved in rearrangement and recombination events including terminal repeat (TR) processing and the EBNA-2 deletion in the EBV strain P3HR-1. Cellular proteins, called terminal or tandem repeat binding proteins (TRBPs), recognize sequences at the junctions of these recombination events. In this study, using antibody supershift assays and expression of recombinant proteins, we show that Sp1 and Sp3 are the sequence specific components of TRBP and that Ku is the nonspecific binding component. Sp1 binds other recombinogenic regions of EBV DNA, but Sp3 does not bind to the large internal repeat. The sequence GGGGTGGGG, a low affinity site for Sp1 and Sp3, is the minimal binding site within terminal repeat binding site 1 (TRBS1). However, 3' flanking sequences in the sequence GGGGTGGGGCATGGGG augment binding of Sp1 and Sp3 so that their affinity of binding is increased approximately twofold relative to a classical high-affinity Sp1 site. EBV lytic cycle induction does not alter the abundance or binding activity of any of the three identified components of TRBP. Sp1 and Sp3 may act in trans to promote EBV terminal repeat processing and possibly other viral and cellular recombination events. PMID- 9344917 TI - The site-specific integration system of the temperate Streptococcus thermophilus bacteriophage phiSfi21. AB - The temperate bacteriophage phiSfi21 integrates its DNA into the chromosome of Streptococcus thermophilus strains via site-specific recombination. Nucleotide sequencing of the attachment sites identified a 40-bp identity region which surprisingly overlaps both the 18-terminal bp of the phage integrase gene and the 11-terminal bp of a host tRNAArg gene. A 2.4-kb phage DNA segment, covering attP, the phage integrase, and a likely immunity gene contained all the genetic information for faithful integration of a nonreplicative plasmid into the attB site. A deletion within the int gene led to the loss of integration proficiency. A number of spontaneous deletions were observed in plasmids containing the 2.4-kb phage DNA segment. The deletion sites were localized to the tRNA side of the identity region and to phage or vector DNA with 3- to 6-bp-long repeats from the border region. A similar type of deletion was previously observed in a spontaneous phage mutant. PMID- 9344918 TI - The 5' region of the human papillomavirus type 31 upstream regulatory region acts as an enhancer which augments viral early expression through the action of YY1. AB - Cis-elements which control human papillomavirus early gene expression have previously been localized to sequences in the upstream regulatory region (URR) which are proximal to the E6 open reading frame. These elements include an enhancer element which functions preferentially in keratinocytes as well as promoter elements. The function of the remaining approximate 500-bp region of the URR in regulating viral expression in the high risk papillomaviruses has been largely uncharacterized. In HPV 6, a negative regulator of early expression, or silencer, has been identified in this 5' region of the URR. In this study, we have investigated the role of the 5' portion of the HPV 31 URR in regulating viral expression. Sequences in this region were found to exert a minimal negative effect on homologous or heterologous promoters. In contrast, a 128-bp sequence within this region was found to exhibit enhancer activity on heterologous and homologous promoters. This enhancer also augmented the activity of the previously identified minimum keratinocyte enhancer. The cellular factors, YY1 and TEF-1, were determined by DNase I footprint analysis and electrophoretic mobility shift assays (EMSAs) to bind the 128-bp enhancer. The binding of YY1 to two of these sites was found to be important for enhancer activity. PMID- 9344919 TI - Failure to complement infectivity of EBV and HSV-1 glycoprotein B (gB) deletion mutants with gBs from different human herpesvirus subfamilies. AB - Glycoprotein B (gB) is conserved among the herpesvirus family which infects a broad range of species. To investigate the functional homology of human alpha herpesviruses, beta-herpesviruses, and gamma-herpesviruses gB proteins, complementation studies were performed with gB genes from each subfamily member using EBV gp110 (EBV gB homologue) and HSV-1 gB null mutants. Neither the alpha herpesvirus HSV-1 gB gene nor the beta-herpesvirus HCMV gB gene were able to complement the gp110 null mutant. Conversely, neither the beta-herpesvirus HCMV gB or the gamma-herpesvirus EBV gp110 gene were able to complement HSV-1 gB null mutants. To further investigate functional domains of EBV gp110 and HSV-1 gB, gB gp110 chimeric proteins were constructed. Surprisingly, none of the chimeric proteins were able to complement either HSV-1 gB null mutants or EBV gp110 null mutants. These results demonstrate that there is not sufficient functional homology between the different gBs to allow complementation in other subfamily members of the herpesvirus family. PMID- 9344964 TI - The three-dimensional hydrodynamics of tadpole locomotion. AB - Tadpoles are unusual among vertebrates in having a globose body with a laterally compressed tail abruptly appended to it. Compared with most teleost fishes, tadpoles swim awkwardly, with waves of relatively high amplitude at both the snout and tail tip. In the present study, we analyze tadpole propulsion using a three-dimensional (3D) computational fluid dynamic (CFD) model of undulatory locomotion that simulates viscous and unsteady flow around an oscillating body of arbitrary 3D geometry. We first confirm results from a previous two-dimensional (2D) study, which suggested that the characteristic shape of tadpoles was closely matched to their unusual kinematics. Specifically, our 3D results reveal that the shape and kinematics of tadpoles collectively produce a small 'dead water' zone between the head-body and tail during swimming precisely where tadpoles can and do grow hind limbs--without those limbs obstructing flow. We next use our CFD model to show that 3D hydrodynamic effects (cross flows) are largely constrained to a small region along the edge of the tail fin. Although this 3D study confirms most of the results of the 2D study, it shows that propulsive (Froude) efficiency for tadpoles is overall lower than predicted from a 2D analysis. This low efficiency is not, however, a result of the high-amplitude undulations of the tadpole. This was demonstrated by forcing our 'virtual' tadpole to swim with fish like kinematics, i.e. with lower-amplitude propulsive waves. That particular simulation yielded a much lower Froude efficiency, confirming that the large amplitude lateral oscillations of the tadpole do, indeed, provide positive thrust. This, we believe, is the first time that the unsteady flow generated by an undulating vertebrate has been realistically modelled in three dimensions. Our study demonstrates the feasibility of using 3D CFD methods to model the locomotion of other undulatory organisms. PMID- 9344920 TI - Adenovirus infection inactivates the translational inhibitors 4E-BP1 and 4E-BP2. AB - Infection with many viruses results in the selective shutoff of host protein synthesis. A common target for virus interference with host protein synthesis is the cap-binding protein complex, eIF4F. The large subunit of the complex, eIF4G, is cleaved upon picornavirus (except cardiovirus) infection. Infection with adenovirus and influenza virus causes dephosphorylation of the cap-binding subunit, eIF4E. Recently, it has been shown that infection with poliovirus or encephalomyocarditis virus activates 4E-BP1, which is a specific inhibitor of eIF4E. Here we show that early in adenovirus infection, 4E-BP1 and its related protein 4E-BP2 are phosphorylated and hence inactivated. This is not consistent with a role of 4E-BPs in adenovirus-induced shutoff, but could explain the increase in protein synthesis reported early in infection. Phosphorylation of 4E BP1 and 4E-BP2 is consistent with earlier findings in adenovirus-infected cells on the activation of the protein kinase p70(S6k), whose phosphorylation lies on the same pathway as 4E-BPs, by E1A. Findings similar to those described here were reported for 4E-BP1 by D. Feigenblum and R. J. Schneider (1996, Mol. Cell. Biol. 16, 5450-5457). PMID- 9344967 TI - Cardiovascular effects of arginine vasotocin in the rainbow trout Oncorhynchus mykiss. AB - The physiological functions of the neurohypophyseal hormone arginine vasotocin (AVT) in teleosts are not clear. In the present studies, the sites and mechanisms of action of AVT on the rainbow trout Oncorhynchus mykiss cardiovascular system were examined in unanesthetized instrumented fish, perfused organs and isolated vessels. Injection of AVT (1, 10 or 100 pmol kg-1 body mass) into trout with dorsal aortic cannulas produced a modest, but dose-dependent, increase in dorsal aortic pressure (PDA). Bolus injection of AVT (100 pmol kg-1 body mass), or continuous infusion (6.7 pmol kg-1 min-1), into trout instrumented with dorsal aortic, ventral aortic and central venous cannulas and a ventral aortic flow probe significantly increased PDA as well as ventral aortic (PVA) and central venous (PVEN) blood pressure. Bradycardia accompanied the rapid rise in PVA while gill resistance (RG) increased. Maximum response to the AVT bolus was reached within 13­21 min and the response decayed slowly over the ensuing 90 min. AVT infusion (6.7 pmol kg-1 min-1) significantly increased PVEN and mean circulatory filling pressure and decreased unstressed blood volume, whereas venous compliance was unaffected. These in vivo studies indicate that AVT increases venous tone, thereby mobilizing blood from the unstressed compartment into the stressed compartment. This increases PVEN, which increases venous return and helps maintain, or slightly elevate, cardiac output. This, combined with an elevated RG and slightly elevated systemic resistance (RS), increases both PVA and PDA; however, the rise in PDA is mitigated by a disproportionate increase in RG relative to RS. In vitro, the effects of AVT are consistent with in vivo responses. AVT increased vascular resistance in the perfused gill and perfused trunk and contracted isolated vascular rings from both rainbow and steelhead trout. The general order of sensitivity of isolated vessels to AVT was (in decreasing order): anterior cardinal vein, celiacomesenteric artery, ductus Cuvier, efferent branchial artery, ventral aorta and coronary artery. Extracellular Ca2+ accounted for over 70 % of the tension in the AVT-contracted efferent branchial artery, but only 57 % of the tension in the anterior cardinal vein. Vascular AVT receptor sensitivity (EC50) in vitro ranged from 0.3 to 6 nmol l-1 and was similar to the estimated ED50 for the dose-dependent increase in PDA in vivo (approximately 1 nmol l-1). AVT was not inotropic in paced ventricular rings nor did it exhibit vasorelaxant activity in perfused organs or vascular rings. These results show that AVT is a potent vasoconstrictor in trout and that its two primary cardiovascular targets are the systemic veins and the branchial vasculature. PMID- 9344969 TI - The steroid pheromone 4-pregnen-17,20ss-diol-3-one increases fertility and paternity in goldfish. AB - Previous studies in goldfish (Carassius auratus) showed that the oocyte maturation-inducing steroid 4-pregnen-17,20ss-diol-3-one (17,20ssP) functions after release as a pheromone that increases male serum gonadotropin II (GtH II) concentration, milt (sperm and seminal fluid) volume and sexual activity, effects hypothesized to increase male reproductive success in the sperm competition of multi-male spawnings. The present study tested this hypothesis by determining whether overnight exposure to 17,20ssP increases fertility. In pair spawnings, 17,20ssP-exposed males fertilized a greater percentage of eggs than did control males, apparently because 17,20ssP-exposed males had more releasable sperm at the onset of spawning. Microsatellite DNA paternity analysis showed that 17,20ssP exposed males also fertilized more eggs in competitive spawnings involving one control male and one 17,20ssP-exposed male. This effect of 17,20ssP on competitive fertility could be due to demonstrated increases in spawning activity, milt volume, duration of sperm motility and proportion of motile sperm. However, it appears that a change in sperm quality is a major component of the pheromonal effect because, in competitive in vitro fertilizations, sperm from 17,20ssP-exposed males fertilized more eggs than did sperm from control males. The results indicate that the response to pheromonal 17,20ssP is a major determinant of reproductive success in male goldfish. PMID- 9344970 TI - Do flatfish feed like other fishes? A comparative study of percomorph prey capture kinematics. AB - The kinematics of prey capture in two bilaterally asymmetrical pleuronectiform flatfish species (Pleuronichthys verticalis and Xystreurys liolepis) and two symmetrical percomorph species (Lepomis macrochirus, a centrarchid, and Cheilinus digrammus, a labrid) were compared to test the hypothesis that flatfish have distinct prey-capture kinematics from those quantified for other percomorph fishes. Size-matched individuals of both flatfish species were video-taped feeding using a high-speed video system. Cephalic displacement and timing variables were quantified and compared with data from similarly sized L. macrochirus and C. digrammus previously collected by other researchers using similar experimental methodology. Nested multivariate analyses of variance indicated that there was no significant difference in prey-capture kinematics between flatfish and non-flatfish taxa, but that prey-capture kinematics did differ among the four taxa. Multiple nested analyses of variance revealed that the taxa differed in 7 of 11 kinematic variables. Post-hoc tests and comparisons with other fish taxa suggest that individuals of P. verticalis possess an unusual combination of prey-capture kinematics including large hyoid depression, large neurocranial rotation, large upper jaw protrusion and small gape. Previous research has suggested that this combination of traits is associated with suction based prey capture. Correspondingly, the ram­suction index calculated for P. verticalis is more negative (indicating a greater use of suction) than that calculated for the other taxa. When homologous kinematic variables are compared across these four taxa, flatfish do not appear to have similar prey-capture kinematics. However, both flatfish species are distinct from the two symmetrical percomorph species in their asymmetrical jaw movements. PMID- 9344973 TI - Work and power output in the hindlimb muscles of Cuban tree frogs Osteopilus septentrionalis during jumping. AB - It has been suggested that small frogs use a catapult mechanism to amplify muscle power production during the takeoff phase of jumping. This conclusion was based on an apparent discrepancy between the power available from the hindlimb muscles and that required during takeoff. The present study provides integrated data on muscle contractile properties, morphology and jumping performance that support this conclusion. We show here that the predicted power output during takeoff in Cuban tree frogs Osteopilus septentrionalis exceeds that available from the muscles by at least sevenfold. We consider the sartorius muscle as representative of the bulk of the hindlimb muscles of these animals, because this muscle has properties typical of other hindlimb muscles of small frogs. At 25 degrees C, this muscle has a maximum shortening velocity (Vmax) of 8.77 +/- 0.62 L0 s-1 (where L0 is the muscle length yielding maximum isometric force), a maximum isometric force (P0) of 24.1 +/- 2.3 N cm-2 and a maximum isotonic power output of 230 +/- 9.2 W kg-1 of muscle (mean +/- S.E.M.). In contrast, the power required to accelerate the animal in the longest jumps measured (approximately 1.4 m) is more than 800 W kg-1 of total hindlimb muscle. The peak instantaneous power is expected to be twice this value. These estimates are probably conservative because the muscles that probably power jumping make up only 85% of the total hindlimb muscle mass. The total mechanical work required of the muscles is high (up to 60 J kg-1), but is within the work capacities predicted for vertebrate skeletal muscle. Clearly, a substantial portion of this work must be performed and stored prior to takeoff to account for the high power output during jumping. Interestingly, muscle work output during jumping is temperature dependent, with greater work being produced at higher temperatures. The thermal dependence of work does not follow from simple muscle properties and instead must reflect the interaction between these properties and the other components of the skeletomuscular system during the propulsive phase of the jump. PMID- 9344975 TI - Acute regulation of glucose uptake in cardiac muscle of the American eel Anguilla rostrata. AB - We investigated the effects of anoxia and contractile activity on glucose uptake and the intracellular location of hexokinase in cardiac muscle of the American eel Anguilla rostrata. Uptake of 2-deoxyglucose (2-DG) by ventricle strips at 15 °C was increased by 45 % by anoxia and by 85 % by contractile activity over basal conditions. The anoxia- and contraction-induced increase in basal 2-DG uptake was inhibited completely by 25 µmol l-1 cytochalasin B, suggesting that facilitated glucose transporters are involved. Maximal activity of hexokinase in whole homogenates (approximately 10 µmol min-1 g-1 tissue) was 200 times higher than the maximal rate of 2-DG uptake measured in vitro (46 nmol min-1 g-1 tissue). Only 20­25 % of hexokinase activity was localized to the mitochondrial fraction, and this was not altered by perfusion of the hearts with anoxic media. It is therefore unlikely that anoxia-induced stimulation of 2 DG uptake is mediated by intracellular translocation of hexokinase. As in the case of mammalian muscle, glucose 6-phosphate is a potent inhibitor of hexokinase in eel cardiac muscle (IC50=0.44 mmol l-1). In summary, anoxia and contractile activity significantly increase 2-DG uptake in cardiac muscle of American eels, and glucose transport may be rate-limiting for glucose utilization. Increased utilization of glucose during anoxia or contractile activity may involve the recruitment of facilitative glucose transport proteins to the cell surface of myocytes or an increase in the intrinsic activity of glucose transporters already residing at the cell surface. PMID- 9344977 TI - Metabolic importance of Na+/K+-ATPase activity during sea urchin development. AB - Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+ ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+ ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development. PMID- 9344979 TI - Fish foot prints: morphology and energetics of the wake behind a continuously swimming mullet (Chelon labrosus Risso). AB - The structure of the wake behind a continuously swimming mullet was analysed qualitatively and quantitatively by applying two-dimensional particle image velocimetry. A detailed analysis of the flow pattern and of the swimming movements of the fish allowed us to derive a kinematic explanation of the flow pattern as well as an estimate of the relative contributions of the body and the tail to thrust production. During active propulsion, the undulatory swimming fish shed a wake consisting in the medio-frontal plane of a rearward, zigzagging jet flow between alternating vortices. The fish shed one vortex per half tailbeat when the tail reached its most lateral position. Part of the circulation shed in the vortices had been generated previously on the body by the transverse body wave travelling down the body. This undulatory pump mechanism accounted for less than half of the energy shed in the wake. The remainder was generated by the tail. The vortex spacing matched the tailbeat amplitude and the stride length. PMID- 9344980 TI - Fatigue of mouse soleus muscle, using the work loop technique. AB - The function of many muscles requires that they perform work. Fatigue of mouse soleus muscle was studied in vitro by subjecting it to repeated work loop cycles. Fatigue resulted in a reduction in force, a slowing of relaxation and in changes in the force-velocity properties of the muscle (indicated by changes in work loop shape). These effects interacted to reduce the positive work and to increase the negative work performed by the muscle, producing a decline in net work. Power output was sustained for longer and more cumulative work was performed with decreasing cycle frequency. However, absolute power output was highest at 5 Hz (the cycle frequency for maximum power output) until power fell below 20% of peak power. As cycle frequency increased, slowing of relaxation had greater effects in reducing the positive work and increasing the negative work performed by the muscle, compared with lower cycle frequencies. PMID- 9344981 TI - Survival of energy failure in the anoxic frog brain: delayed release of glutamate. AB - This study investigated the relationship between energy failure and neurotransmitter release in the frog (Rana pipiens) brain during 1-3 h of anoxia. Unlike truly anoxia-tolerant species, the frog does not defend its brain energy charge. When exposed to anoxia at 25 degrees C, there is an immediate fall in brain ATP levels, which reach approximately 20% of normoxic levels in approximately 60 min. The frog, nevertheless, survives another 1-2 h of anoxia. At 100 min of anoxia, there is an increase in extracellular adenosine concentration, probably originating from the increased intracellular adenosine concentration caused by the breakdown of intracellular ATP. Increases in the levels of extracellular glutamate and GABA do not occur until 1-2 h after ATP depletion. This response is quite unlike that recorded for other vertebrates, anoxia-tolerant or anoxia-intolerant, where energy failure quickly results in an uncontrolled and neurotoxic release of excitatory neurotransmitters. In the frog, the delay in excitotoxic neurotransmitter release may be one of the factors that allow a period of survival after energy failure. Clearly, energy failure by itself is not a fatal event in the frog brain. PMID- 9344982 TI - Voltage sensing in jellyfish Shaker K+ channels. AB - The S4 segment of the jellyfish (Polyorchis penicillatus) Shaker channel jShak1 contains only six positively charged motifs. All other Shaker channels, including the jellyfish Shaker channel jShak2, have seven charges in this segment. Despite their charge differences, both these jellyfish channels produce currents with activation and inactivation curves shifted by approximately +40 mV relative to other Shaker currents. Adding charge without changing segment length by mutating the N-terminal side of jShak1 S4 does not have a pronounced effect on channel activation properties. Adding the positively charged motif RIF on the N-terminal side of K294 (the homologue of K374 in Drosophila Shaker, which is a structurally critical residue) produced a large positive shift in both activation and inactivation without altering the slope of the activation curve of the channel. When IFR was added to the other side of K294, there was a small negative shift in activation and fast inactivation of the channel was prevented. Our results demonstrate that K294 divides the S4 segment into functionally different regions and that the voltage threshold for activation and inactivation of the channel is not determined by the total charge on S4. PMID- 9345009 TI - Dendritic cells: unique leukocyte populations which control the primary immune response. PMID- 9345010 TI - Cytoplasmic localization of a functionally active Fanconi anemia group A-green fluorescent protein chimera in human 293 cells. AB - Hypersensitivity to cross-linking agents and predisposition to malignancy are characteristic of the genetically heterogeneous inherited bone marrow failure syndrome, Fanconi anemia (FA). The protein encoded by the recently cloned FA complementation group A gene, FAA, has been expected to localize in the nucleus as based on the presence of sequences homologous to a bipartite nuclear localization signal (NLS) and a leucine repeat motif. In contrast to this expectation, we show here that a functionally active FAA-green fluorescent protein (GFP) hybrid resides in the cytoplasmic compartment of human kidney 293 cells. In accordance with this finding, disruption of the putative NLS by site directed mutagenesis failed to affect both subcellular localization and the capacity to complement hypersensitivity to the cross-linking agent mitomycin C in FA-A lymphoblasts. Furthermore, the N-terminal part of FAA with the putative NLS at amino acid position 18 to 35 showed no nuclear translocation activity when fused to GFP, while the first 115 N-terminal amino acids appeared to be indispensable for the complementing activity in FA-A cells. Similarly, mutagenesis studies of the putative leucine repeat showed that, even though this region of the protein is important for complementing activity, this activity does not depend on an intact leucine zipper motif. Finally, fusion of the NLS motif derived from the SV40 large T antigen to FAA could not direct the hybrid protein into the nucleus of 293 cells, suggesting that FAA is somehow maintained in the cytoplasm via currently unknown mechanisms. Thus, like the first identified FA protein, FAC, FAA seems to exert its function in the cytoplasmic compartment suggesting FA proteins to be active in a yet to be elucidated cytoplasmic pathway that governs hematopoiesis and protects against genomic instability. PMID- 9345011 TI - Functional correction of Fanconi anemia group A hematopoietic cells by retroviral gene transfer. AB - Fanconi anemia (FA) is an autosomal recessive genetic disorder characterized by a variety of physical anomalies, bone marrow failure, and an increased risk for malignancy. FA cells exhibit chromosomal instability and are hypersensitive to DNA cross-linking agents such as mitomycin C (MMC). FA is a clinically heterogeneous disorder and can be functionally divided into at least five different complementation groups (A-E). We previously described the use of a retroviral vector expressing the FAC cDNA in the complementation of mutant hematopoietic cells from FA-C patients. This vector is currently being tested in a clinical trial of ex vivo hematopoietic progenitor cell transduction. The FA-A group accounts for over 65% of all FA cases, and the FAA cDNA was recently identified by both expression and positional cloning techniques. We report here the transduction and phenotypic correction of lymphoblastoid cell lines from four unrelated FA-A patients, using two amphotropic FAA retroviral vectors. Expression of the FAA transgene was adequate to normalize cell growth, cell-cycle kinetics, and chromosomal breakage in the presence of MMC. We then analyzed the effect of retroviral vector transduction on hematopoietic progenitor cell growth. After FAA transduction of mutant progenitor cells, either colony number or colony size increased in the presence of MMC. In addition, FAA but not FAC retroviral transduction markedly improved colony growth of progenitor cells derived from an unclassified FA patient. FAA retroviral vectors should be useful for both complementation studies and clinical trials of gene transduction. PMID- 9345012 TI - Enhanced green fluorescent protein as selectable marker of retroviral-mediated gene transfer in immature hematopoietic bone marrow cells. AB - The further improvement of gene transfer into hematopoietic stem cells and their direct progeny will be greatly facilitated by markers that allow rapid detection and efficient selection of successfully transduced cells. For this purpose, a retroviral vector was designed and tested encoding a recombinant version of the Aequorea victoria green fluorescent protein that is enhanced for high-level expression in mammalian cells (EGFP). Murine cell lines (NIH 3T3, Rat2) and bone marrow cells transduced with this retroviral vector demonstrated a stable green fluorescence signal readily detectable by flow cytometry. Functional analysis of the retrovirally transduced bone marrow cells showed EGFP expression in in vitro clonogenic progenitors (GM-CFU), day 13 colony-forming unit-spleen (CFU-S), and in peripheral blood cells and marrow repopulating cells of transplanted mice. In conjunction with fluorescence-activated cell sorting (FACS) techniques EGFP expression could be used as a marker to select for greater than 95% pure populations of transduced cells and to phenotypically define the transduced cells using antibodies directed against specific cell-surface antigens. Detrimental effects of EGFP expression were not observed: fluorescence intensity appeared to be stable and hematopoietic cell growth was not impaired. The data show the feasibility of using EGFP as a convenient and rapid reporter to monitor retroviral-mediated gene transfer and expression in hematopoietic cells, to select for the genetically modified cells, and to track these cells and their progeny both in vitro and in vivo. PMID- 9345013 TI - High-titer retroviral vectors containing the enhanced green fluorescent protein gene for efficient expression in hematopoietic cells. AB - Retroviral vectors constitute the most efficient system to deliver and integrate foreign genes into mammalian cells. We have developed a producer cell line that yields high titers of amphotropic retroviral vectors carrying the enhanced green fluorescent protein (EGFP) gene, a codon humanized, red-shifted variant of the green fluorescent protein (GFP) gene, which can be used as a selectable marker. We have used a hybrid vector that has been shown to efficiently drive gene expression in hematopoietic cells. Virtually all murine and human cell lines and primary human hematopoietic cells tested were transduced with varying efficiency after incubation with vector-containing supernatants. Human CD34(+) cells obtained from cord blood or aphereses products were transduced using a protocol that involves daily addition of vector-containing supernatants for 6 consecutive days. At day 6, up to 16% of the cells expressed EGFP, as assessed by flow cytometry. Sorted EGFP-expressing cells were able to produce fluorescent hematopoietic colonies. EGFP's main advantages are its fast flow cytometry determination and the possibility of cell sorting and simultaneous evaluation of the transduction efficiency along with other phenotypic markers. PMID- 9345014 TI - Induction of vascular endothelial growth factor by hypoxia is modulated by a phosphatidylinositol 3-kinase/Akt signaling pathway in Ha-ras-transformed cells through a hypoxia inducible factor-1 transcriptional element. AB - Tumor angiogenesis, the development of new blood vessels, is a highly regulated process that is controlled genetically by alterations in oncogene and tumor suppressor gene expression and physiologically by the tumor microenvironment. Previous studies indicate that the angiogenic switch in Ras-transformed cells may be physiologically promoted by the tumor microenvironment through the induction of the angiogenic mitogen, vascular endothelial growth factor (VEGF). In this report, we show Ras-transformed cells do not use the downstream effectors c-Raf-1 or mitogen activated protein kinases (MAPK) in signaling VEGF induction by hypoxia as overexpression of kinase-defective alleles of these genes does not inhibit VEGF induction under low oxygen conditions. In contrast to the c-Raf 1/MAP kinase pathway, hypoxia increases phosphatidylinositol 3-kinase (PI 3 kinase) activity in a Ras-dependent manner, and inhibition of PI 3-kinase activity genetically and pharmacologically results in inhibition of VEGF induction. We propose that hypoxia modulates VEGF induction in Ras-transformed cells through the activation of a stress inducible PI 3-kinase/Akt pathway and the hypoxia inducible factor-1 (HIF-1) transcriptional response element. PMID- 9345015 TI - Transcriptional behavior of LCR enhancer elements integrated at the same chromosomal locus by recombinase-mediated cassette exchange. AB - Efficient integration of transgenes at preselected chromosomal locations was achieved in mammalian cells by recombinase-mediated-cassette-exchange (RMCE), a novel procedure that makes use of the CRE recombinase together with Lox sites bearing different spacer regions. We have applied RMCE to the study of the human beta-globin gene Locus Control Region by integrating at the same genetic locus in MEL cells, a LacZ gene driven by the human beta-globin promoter linked to HS2 and HS3 alone or in combination with HS4. Expression studies at the cell population level and in individual cells before and after induction of differentiation with hemin or DMSO show that the presence of these enhancers is associated with variegated patterns of expression. We were able to show that the LCR fragments tested act by controlling both the probability of expression and the rate of transcription of the linked beta-globin promoter. Both of these factors were also dependent on the state of differentiation of the MELc and on the presence of a second transcription unit located in cis. The ability to manipulate by RMCE constructs integrated into chromosomes should help in the creation of complex, rationally designed, artificial genetic loci. PMID- 9345016 TI - Induction of apoptosis without differentiation by retinoic acid in PLB-985 cells requires the activation of both RAR and RXR. AB - Retinoic acid (RA) induces differentiation, followed by apoptosis in acute promyelocytic leukemia (APL) cells, both in vitro and in patients. One problem in understanding these mechanisms is to distinguish molecular events leading to differentiation from those leading to apoptosis. We have identified a leukemic cell line, PLB-985, where RA directly induces apoptosis with no morphologic, genetic, or cell-surface marker evidence of differentiation. These cells differentiate following dimethyl sulfoxide (DMSO), but not RA, treatment. Two color flow cytometry showed no alteration of the cell cycle after RA treatment, and cell-surface marker analysis of CD11a, CD11b, and CD13 showed no modulation typical of differentiating cells. RNA expression of myeloblastin and transglutaminase, genes regulated by RA-induced differentiation in NB4 cells, was unchanged by RA treatment. Instead, RA induced apoptosis, as shown by typical apoptotic morphological features, genomic DNA laddering, and positive labeling in the TUNEL assay. We found that induction of apoptosis in this model requires a different pattern of retinoid receptor binding and transcriptional activation than is seen in APL cells. As previously described, treatment with retinoid receptor-selective ligands showed that stimulation of RAR alone is sufficient to induce differentiation and apoptosis in NB4 cells, and that stimulation of RXR has no effect on the parameters analyzed. In PLB-985 cells, on the other hand, apoptosis was induced only upon costimulation of both RAR and RXR. Stimulation of either receptor alone had no effect on the cells. Consistent with these findings, bcl-2 RNA and protein levels were downregulated after stimulation of both RAR and RXR, but not with an RAR-specific ligand alone, as in NB4 cells. The expression of several other bcl-2 family members (bcl-X, ich-1, bax, bag, and bak ) and retinoid receptors (RARalpha, RXRalpha, and RXRbeta) was not affected by treatment with RAR- and/or RXR-activating retinoids; RARbeta RNA was undetectable before and after retinoid treatment. Thus, our cell model provides a useful tool in determining the genetic events mediating apoptosis as a response to RA, unobscured by events implicated in differentiation. PMID- 9345017 TI - Interleukin-10 counterregulates proinflammatory cytokine-induced inhibition of neutrophil apoptosis during severe sepsis. AB - Neutrophils play a key role in the pathophysiology of septic multiple organ dysfunction syndrome (MODS) through excessive release of toxic granule components and reactive oxygen metabolites with consequent tissue destruction. The increase of senescent neutrophils during sepsis indicates a potential breakdown of autoregulatory mechanisms including apoptotic processes to remove activated neutrophils from inflammatory sites. Therefore, neutrophil apoptosis of patients with severe sepsis and its regulatory mechanisms were investigated. Spontaneous neutrophil apoptosis from patients with severe sepsis was significantly reduced in comparison to healthy individuals. Cytokines detected in the circulation during sepsis (tumor necrosis factor-alpha [TNF-alpha], interferon-gamma [IFN gamma], granulocyte colony-stimulating factor [G-CSF], granulocyte-macrophage colony-stimulating factor [GM-CSF]) inhibited neutrophil apoptosis in both groups, though the effect was more distinct in neutrophils from healthy humans. Addition of lipopolysaccharide (LPS) to neutrophils from healthy humans markedly (P < .05) reduced apoptosis which was partially restored through addition of anti TNF-antibody. Interleukin-10 (IL-10) counteracted (P < .05) inhibition of neutrophil apoptosis induced by LPS, recombinant human (rh) TNF-alpha, rhIFN gamma, rhG-CSF, and rhGM-CSF, whereas rhIL-4 or rhIL-13 were ineffective. Reduced neutrophil apoptosis during sepsis was concomitant with increased tyrosine phosphorylation, while IL-10 markedly inhibited tyrosine phosphorylation in LPS stimulated neutrophils. These results identify proinflammatory cytokines and IL 10 as strong regulators of spontaneous neutrophil apoptosis during sepsis. Inhibition as well as acceleration of neutrophil apoptosis seems to be associated with alterations of signal transduction pathways. PMID- 9345019 TI - Treatment of polycythemia vera: the use of hydroxyurea and pipobroman in 292 patients under the age of 65 years. AB - Nonradiomimetic drugs, hydroxyurea (HU) and pipobroman (Pi), were administred to relatively young subjects with polycythemia vera (PV) in an attempt to decrease the leukemogenic risk observed in patients treated with 32P. Clinical safety, hematological efficacy, risk of carcinoma or leukemia, and frequency of progression to myelofibrosis have not yet been defined in long-term studies, and no comparative studies of HU and Pi have been conducted. Since 1980, 292 patients with PV diagnosed before the age of 65 years were randomized to receive treatment with HU (25 mg/kg/d, followed by low-dose maintenance) or Pi (1.2 mg/kg/d, followed by low-dose maintenance). Patients were followed until death or until May 1997. Drug tolerance was often poor; leg ulcers and buccal aphthous ulcers (with HU) and gastric pain and diarrhea (with Pi) sometimes required treatment change, mainly in the HU arm. Hematological stability, especially in terms of platelet count, was very often insufficient with HU (45% of cases), but the risk of thrombo-embolic event was similar in both arms. Actuarial survival was similar in the two arms and shorter than that of the reference population. The risk of leukemia was approximately 10% at the 13th year, with no significant difference between the two arms. The risk of carcinoma (when excluding the skin cancers) was similar in both groups. There was a high risk of progression to myelofibrosis in the patients treated by HU, which was significantly higher than with Pi. PMID- 9345018 TI - Stimulation of hematopoiesis by amifostine in patients with myelodysplastic syndrome. AB - The aminothiol, amifostine (Ethyol; U.S. Bioscience, West Conshohocken, PA), is a cytoprotective agent that ameliorates the toxicities of anticancer therapy. In vitro, amifostine promotes the formation and survival of primitive hematopoietic progenitors derived from myelodysplastic bone marrow (BM) specimens. To evaluate the hematological effects of amifostine, 18 patients with myelodysplastic syndrome (MDS) and one or more refractory cytopenias received treatment with amifostine in a Phase I/II study. Four cohorts received intravenous treatment with 100, 200, or 400 mg/m2 amifostine three times a week, or 740 mg/m2 weekly for three consecutive weeks followed by 2 weeks observation. Nonresponding patients received a second course of therapy at the next higher dose level depending upon drug tolerance. Bone marrow (BM) progenitor growth was assessed before treatment and after day 21. Diagnoses included refractory anemia (7), refractory anemia with ringed sideroblasts (5), refractory anemia with excess blasts (RAEB) (4), and RAEB-in transformation (RAEB-t) (2). Single- or multi lineage hematologic responses occurred in 15 patients (83%) treated with the three-times-a-week dose schedule. Fourteen patients had a 50% or greater increase in absolute neutrophil count with amifostine treatment (range, 426 to 11,348/microL). Platelet count increased in 6 (43%) of 14 patients with thrombocytopenia (absolute increase, 16, 000 to 110,000/microL), and 5 of 15 red blood cell transfusion-dependent patients had a 50% of greater reduction in transfusion needs. Assayable hematopoietic progenitors increased in 13 of 15 evaluable patients; including CFU-GEMM (12), BFU-E (8), and CFU-GM (6). Amifostine doses less than or equal to 200 mg/m2 were well tolerated, whereas grade II nausea, vomiting, and fatigue was limiting at higher doses. Three patients with excess blasts before enrollment experienced an increase in BM blast percentage and two patients had evolution to acute leukemia that persisted after treatment withdrawal. We conclude that amifostine administered at doses 4 months) transplanted with bone marrow cells transduced with high-titer virus. Southern blotting confirms the presence of the unrearranged 5.1-kb human beta-globin gene-containing provirus in 2 of these mice. In addition, long-term expression of the transferred gene is seen in 2 mice at levels of 5% and 20% that of endogenous murine beta-globin at 6 and 8 months posttransplantation. We further document stem cell transduction by the successful transfer and high-level expression of the human beta-globin gene from mice transduced 9 months earlier into irradiated secondary recipient mice. These results demonstrate high-level, long-term somatic human beta-globin gene transfer into the hematopoietic stem cells of an animal for the first time, and suggest the potential feasibility of a retroviral gene therapy approach to sickle cell disease and the beta thalassemias. PMID- 9345025 TI - Normal platelets and megakaryocytes are produced in vivo in the absence of thrombopoietin. AB - Thrombopoietin (TPO) has been established as the major regulator of megakaryocyte and platelet production. In vitro and in vivo studies have demonstrated that TPO affects both megakaryocyte proliferation and maturation. In vitro, TPO has been reported to be essential for full development of megakaryocytes and platelets. These studies are in contrast to results observed in vivo in mice deficient in the TPO or c-mpl gene (TPO-/- and c-mpl-/-). Both TPO-/- and c-mpl-/- mice exhibit a 90% reduction in megakaryocyte and platelet levels. But even with this small number of circulating platelets, these mice do not have any excessive bleeding. Ultrastructural analysis indicates that platelets and megakaryocytes present in the knockout mice are morphologically normal. Characterization of platelet function shows that platelets from knockout mice are functionally identical to the wild-type platelets as measured by upregulation of 125I fibrinogen binding to platelets in response to adenosine diphosphate (ADP) stimulation and by platelet attachment to the immobilized extracellular matrix proteins, collagen and von Willebrand factor (vWF). These results demonstrate that in vivo, TPO is required for the control of megakaryocyte and platelet number but not for their maturation. Other factors with megakaryocytopoietic activity may be able to compensate for the maturational role of TPO and lead to the formation of normal megakaryocytes and platelets in TPO-/- and c-mpl-/- mice. PMID- 9345026 TI - Complex regulation of CDK2 during phorbol ester-induced hematopoietic differentiation. AB - Phorbol myristate acetate (PMA) treatment of U937 human leukemic cells results in late G1 cell cycle arrest and terminal monocyte/macrophage-like differentiation. The PMA-induced G1 arrest involves a marked decrease in cdk2 activity, which correlates with total cdk2 dephosphorylation. Here, we show that the levels of cyclin A mRNA and protein markedly decrease during PMA-induced differentiation of U937 cells. In contrast, the level of cyclin E protein remains unchanged and in a complex with cdk2 during the entire course of PMA treatment. During the PMA induced differentiation, cyclin E-associated cdk2 activity drops markedly. Furthermore, the amount of p27(Kip1) protein associated with cyclin E/cdk2 greatly increases 24 to 72 hours after PMA treatment. The absence of changes in p27(Kip1) mRNA levels by Northern blot suggest that the levels of this protein are controlled by posttranscriptional or posttranslational mechanism(s). These results show that the mechanisms mediating PMA-induced G1 arrest are complex. The inhibition of cdk2 activity is associated with (1) a decrease in cyclin A protein levels, (2) inactivation of cdk2 complexes, and (3) upregulation of p27(Kip1) protein. PMID- 9345027 TI - Leptin stimulates the proliferation of murine myelocytic and primitive hematopoietic progenitor cells. AB - Leptin, the product of obese gene, was originally identified as a factor regulating body-weight homeostasis and energy balance. The present study has shown that leptin acts on murine hematopoiesis in vitro. In the culture of bone marrow cells (BMC) of normal mice, leptin induced only granulocyte-macrophage (GM) colony formation in a dose-dependent manner, and no other types of colonies were detected even in the presence of erythropoietin (Epo). Leptin also induced GM colony formation from BMC of db/db mutant mice whose leptin receptors were incomplete, but the responsiveness was significantly reduced. The effect of leptin on GM colony formation from BMC of normal mice was also observed in serum free culture, and comparable with that of GM-colony-stimulating factor (CSF ). Although leptin alone supported few colonies from BMC of 5-fluorouracil (5-FU) treated mice in serum-free culture, remarkable synergism between leptin and stem cell factor (SCF ) was obtained in the colony formation. The addition of leptin to SCF enhanced the SCF-dependent GM colony formation and induced the generation of a number of multilineage colonies in the presence of Epo. When lineage (Lin) Sca-1(+) cells sorted from BMC of 5-FU-treated mice were incubated in serum-free culture, leptin synergized with SCF in the formation of blast cell colonies, which efficiently produced secondary colonies including a large proportion of multilineage colonies in the replating experiment. In serum-free cultures of clone-sorted Lin-c-Kit+Sca-1(+) and Lin-c-Kit+Sca-1(-) cells, although synergism of leptin and SCF was observed in the colony formation from both cells, leptin alone induced the colony formation from Lin-c-Kit+Sca-1(-), but not Lin-c-Kit+Sca 1(+) cells. These results have shown that leptin stimulates the proliferation of murine myelocytic progenitor cells and synergizes with SCF in the proliferation of primitive hematopoietic progenitors in vitro. PMID- 9345028 TI - Thrombopoietin is synthesized by bone marrow stromal cells. AB - We have previously characterized stromal progenitor cells contained in fetal bone marrow by fluorescence-activated cell sorting (FACS) using the differential expression of CD34, CD38, and HLA-DR, and found that a small number were contained within the CD34(+) cell fraction. In the present study, the frequency of stromal progenitors in both the CD34+ and CD34- subpopulations from samples of fetal and adult bone marrow was approximately one in 5,000 of the mononuclear cell fraction. Using multiparameter single-cell sorting, one in 20 fetal bone marrow cells with the CD34+, CD38-, HLA-DR-, CDw90+ phenotype were clonogenic stromal progenitors, whereas greater than one in five single cells with the CD34 , CD38-, HLA-DR-, CDw90+ phenotype formed stromal cultures. We found that cultures initiated by hematopoietic and stromal progenitors contained within the CD34+ fraction of bone marrow cells formed mixed hematopoietic/stromal cell cultures that maintained the viability of the hematopoietic progenitor cells for 3 weeks in the absence of added hematopoietic cytokines. We characterized some of the hematopoietic cytokines synthesized by stromal cultures derived from either CD34+ or CD34- bone marrow cells using reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of interleukin-3 (IL-3), stem cell factor (SCF), CD34, Flt3/Flk2 ligand (FL), and thrombopoietin (TPO) mRNA sequences. We found ubiquitous expression of TPO mRNA in greater than 90% of stromal cultures initiated by either CD34+ or CD34- cells, and variable expression of SCF, FL, and CD34 mRNA. In particular, SCF and CD34 mRNA were detected only in stromal cultures initiated by CD34+ bone marrow cells, although the differences between CD34+ and CD34- stromal cells were not statistically significant. IL-3 mRNA was not found in any stromal cultures. An enzyme-linked immunosorbent assay (ELISA) of soluble SCF and TPO present in culture supernatants demonstrated that biologically significant amounts of protein were secreted by some cultured stromal cells: eight of 16 samples of conditioned media from stromal cultures initiated by fetal and adult bone marrow contained more than 32 pg/mL SCF (in the linear range of the ELISA), with a median value of 32 pg/mL (range, 9 to 230), while 13 of 24 samples of conditioned media had more than 16 pg/mL TPO (in the linear range of the ELISA), with a median of 37 pg/mL (range, 16 to 106). Our data indicate that stromal cultures initiated by single bone marrow cells can make FL, SCF, and TPO. Local production of early-acting cytokines and TPO by stromal cells may be relevant to the regulation of hematopoietic stem cell self renewal and megakaryocytopoiesis in the bone marrow microenvironment. PMID- 9345029 TI - Chronic thrombocytopenia is induced in dogs by development of cross-reacting antibodies to the MpL ligand. AB - The MpL ligand (ML) is a potent stimulus for thrombocytopoiesis. To create an in vivo model of ML deficiency, we injected dogs with a recombinant human ML (rhML) to determine whether cross-reacting antibodies would develop and cause thrombocytopenia. RhML was administered subcutaneously for 8 weeks to three normal dogs (mean platelets, 197 +/- 5.5 x 10(3)/microL). Within 5 days their platelet counts were twice baseline and greater than 4 times baseline by day 21. Then, uniformly, chronic thrombocytopenia developed. At 1 week after terminating rhML, mean platelets were 0.5 times baseline and at 2 months 0.25 times baseline. Early in treatment, marrow biopsies showed increased megakaryocyte number and ploidy, which decreased as platelets declined. Paralleling these changes, high titer anti-rhML antibodies developed. Autologous 51Cr-labeled platelet recovery and survival measurements indicated that the thrombocytopenia was principally due to decreased production. Infusion of plasma from the thrombocytopenic dogs into two normal dogs and one dog previously made thrombocytopenic with rhML caused platelet counts to fall gradually. These studies show that dogs with anti-rhML antibodies develop thrombocytopenia, presumably because the cross-reacting antibodies neutralize endogenous canine ML. The results strongly suggest that ML plays an essential role in maintaining normal platelet levels. PMID- 9345030 TI - Activation of the mitogen-activated protein kinase pathway is involved in and sufficient for megakaryocytic differentiation of CMK cells. AB - Activation of the mitogen-activated protein (MAP) kinase pathway has been associated with both cell proliferation and differentiation. Constitutively activated forms of Mek (MAP kinase/Erk kinase) and Erk (MAP kinase) have been previously shown capable of inducing differentiation or proliferation in nonhematopoietic cells. To specifically examine the role of Erk activation in megakaryocytic growth and development, we activated the MAP kinase pathway by the transfection of constitutively activated Mek or Erk cDNA into a human megakaryoblastic cell line, CMK, by electroporation. The CMK transfectant clones that expressed constitutively activated Mek or Erk showed morphologic changes of differentiation. Transfected cells also showed expression of mature megakaryocytic cell surface markers. The MAP kinase pathway was also activated by treatment of the hematopoietic cells with a cytokine that activates Erk. The treatment of CMK cells with stem cell factor (SCF ) caused MAP kinase activation and induced differentiation by the expression of mature megakaryocytic cell surface markers. The effects of the SCF treatment were inhibited by pretreatment with a specific inhibitor of the MAP kinase pathway, PD98059. In this report, we conclude that activation of the MAP kinase pathway was both necessary and sufficient to induce differentiation in this megakaryoblastic cell line. PMID- 9345031 TI - Characterization of stromal progenitor cells enriched by flow cytometry. AB - The progenitors for cells of bone, cartilage, fat, and muscle are thought to be derived from mesenchymal stem cells but despite extensive study of stromal cell differentiation, neither mesenchymal stem cells or the more committed, tissue specific progenitors have been well-characterized. In this study we used flow cytometry to isolate from fetal rat periosteum a population of small, slowly cycling cells with low cytoplasmic granularity (S cells) that display stem cell characteristics. On plating, S cells exhibited a 90% higher labeling index with [3H]-thymidine compared to unsorted cells and when grown in culture generated cartilage, adipocyte, and smooth muscle phenotypes, in addition to bone. Only the S-cell population showed extensive self-renewal of cells with osteogenic potential. Electron microscopy showed that S cells have high nuclear:cytoplasmic ratios with large condensed nuclei and a paucity of cytoplasmic organelles. Freshly sorted suspensions of immunocytochemically stained S cells did not express differentiation-associated markers such as type I, II, and III collagens, alkaline phosphatase, or osteopontin. However, after attachment, S cells became immunopositive for collagens I, II, III, osteopontin, and also for the cell surface receptor CD44, which mediates cell attachment to hyaluronan and osteopontin. These studies show that viable osteogenic precursor cells with the stem cell characteristics of self-renewal, high proliferative capacity, and multipotentiality can be enriched from heterogeneous stromal cell populations with simple flow cytometric methods. These cells may be useful for regeneration of stromal tissues. PMID- 9345032 TI - Generation of virtually pure and potentially proliferating dendritic cells from non-CD34 apheresis cells from patients with multiple myeloma. AB - Defects in immune response are often reported in patients with multiple myeloma (MM). Because dendritic cells (DCs) are key effectors in promoting cellular immunity and are potential vectors for immunotherapy, we have evaluated the ability of MM patients' apheresis cells to generate DCs in short-term cultures. We report here the obtaining of a virtually pure population of DCs (89.7% +/- 6%, n = 18) after culturing adherent apheresis cells for 7 days with granulocyte macrophage colony-stimulating factor (GM-CSF ) and interleukin-4 (IL-4). These cells exhibited all the phenotypic characteristics (CD1a+, HLA-DR+, CD80+, CD40+, CD14-) and the MLR stimulating capacity of mature DCs. The number of DCs reached 12. 1% of the initial apheresis cell number put into culture. As DC precursors involved in this model were CD34(-) cells, the unabsorbed cells resulting from clinical-grade CD34 purification were a reliable source of DCs, even after freezing. The proliferation of DC precursors could be increased 10-fold by adding IL-3 and tumor necrosis factor-alpha together with GM-CSF and IL-4. Thus, CD34- apheresis cells from patients with MM offer an interesting source for generating pure, functional, and potentially proliferating DCs. PMID- 9345033 TI - Successful reconstitution of human hematopoiesis in the SCID-hu mouse by genetically modified, highly enriched progenitors isolated from fetal liver. AB - Highly purified CD34++CD38-Lin- hematopoietic progenitors isolated from human fetal liver were infected with the murine retroviral vector, MFG nls-LacZ, which encodes a modified version of the Escherichia coli beta-galactosidase gene. Progenitors that were cocultured with the packaging cell line could reconstitute human bone marrow or thymus implanted in SCID-hu mice. Expression of the beta galactosidase gene was observed in primitive and committed clonogenic progenitors, mature myeloid, B-lineage cells, and T-lineage cells for up to 4 months after injection into SCID-hu mice. Furthermore, hematopoietic reconstitution by genetically modified progenitor cells could be achieved by the injection of the cells generated from as few as 500 CD34++CD38-Lin- cells, suggesting efficient retroviral gene transfer into fetal liver progenitors. PMID- 9345034 TI - Signal transduction in human hematopoietic cells by vascular endothelial growth factor related protein, a novel ligand for the FLT4 receptor. AB - We have recently identified a novel ligand of the vascular endothelial growth factor (VEGF) family termed VEGF-related protein (VRP), which specifically binds to the FLT4 receptor. To characterize the signaling events after VRP engagement of its cognate receptor in hematopoietic cells, a population of human erythroleukemia (HEL) cells, termed HEL-JW, expressing high levels of FLT4 receptor was isolated. Stimulation of HEL-JW cells with VRP alone and in combination with the c-kit ligand/stem cell factor increased cell growth. VRP induced tyrosine phosphorylation of various proteins, including the FLT4 receptor. Further characterization of these tyrosine phosphorylated molecules revealed that Shc, Grb2, and SOS form a complex with the activated FLT4 receptor. HEL-JW cells also expressed RAFTK, a recently identified member of the focal adhesion kinase family. RAFTK was phosphorylated and activated upon VRP treatment, and there was an enhanced association of this kinase with the adaptor protein Grb2. Furthermore, the c-Jun NH2-terminal kinase (JNK), involved in growth activation and shown to mediate RAFTK signaling in other cell types, was activated by VRP stimulation. We also observed that VRP treatment of HEL-JW cells resulted in the phosphorylation of the cytoskeletal protein paxillin. This treatment resulted in an increased association of paxillin with RAFTK, which was mediated by the C-terminal region of RAFTK. These studies indicate that VRP stimulation induced the formation of a signaling complex at its activated receptor as well as activation of RAFTK. VRP-mediated activation of RAFTK may facilitate signal transduction to the cytoskeleton and downstream to the JNK pathway in FLT4-expressing blood cells. PMID- 9345035 TI - Development of osteoclasts from embryonic stem cells through a pathway that is c fms but not c-kit dependent. AB - Osteoclasts are hematopoietic cells essential for bone resorption. To study the derivation of these interesting cells, we developed a stepwise culture system where stromal cells promote embryonic stem (ES) cells to differentiate into mature osteoclasts. Three phases to this differentiation process include (1) induction of hematopoiesis, along with the generation of osteoclast precursors, (2) expansion of these precursors, and (3) terminal differentiation into mature osteoclasts in the presence of 1alpha,25-dihydroxyvitamine D3 . Although the transition of ES cells to the hematopoietic lineage was not blocked by an antibody to c-fms, later phases were dependent on a signaling through this transmembrane receptor as indicated by the finding that anti-c-fms treatment of cells in the second and third phases reduced the number of osteoclasts produced by 75% and more than 99%, respectively. Blockade of signaling through another tyrosine kinase-type receptor, c-kit, did not affect any stages of osteoclastogenesis, although generation of other hemopoietic lineages was reduced to less than 10% of untreated. When small numbers of ES cells were directly cultured under conditions that promote osteoclast differentiation, tartrate resistant acid phosphatase-positive multinucleated cells were observed at the edge but not inside of colonies. This suggests that some types of cell-cell interactions may inhibit development of mature osteoclasts. The culture system developed here provides an important tool for osteoclast biology. PMID- 9345036 TI - Adhesion and migration are differentially regulated in hematopoietic progenitor cells by cytokines and extracellular matrix. AB - The conditions that control the migratory status of hematopoietic progenitor cells on extracellular matrix (ECM) and that decide whether a cell migrates or adheres are incompletely understood. We analyzed the migratory behavior of murine hematopoietic progenitor cells factor-dependent-cell-paterson (FDCP)-mix and purified lin-Sca1+ bone marrow cells on ECM. We found that migration on fibronectin (Fn) or laminin (Lam) becomes dependent on beta1-integrins if a surface restraint force is introduced by tilting the ECM-coated culture vessels. Under these conditions, migration specifically occured on Fn and Lam, and was not detected on collagen IV-, hyaluronate-, or bovine serum albumin- coated surfaces. Migration depended on the continuous presence of hematopoietic cytokines interleukin-3 (IL-3), granulocyte colony-stimulating factor (G-CSF), macrophage CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF), or stem cell factor (SCF), whereas other cytokines, such as IL-8, macrophage inflammatory protein-1alpha, macrophage-chemotactic and activating factor, and erythropoietin resulted in very little or no migratory response. IL-3 induced migration was synergistically enhanced by other CSFs, but was completely inhibited by addition of transforming growth factor-beta1. In contrast to firm local adhesion of previously cytokine depleted progenitors that was rapidly inducible within 1 hour after exposure to cytokines, preincubation on Fn matrix for 4 to 6 hours was required before cytokines could induce migration. A sudden increase of cytokine concentration reversibly inhibited migration and induced a fully adhesive state; this effect could be prolonged by consecutive stimulation with heterologous cytokines. Whereas cytokines activated resting progenitor cells to migrate on ECM, cell migration speed was regulated by Fn concentration. These results indicate that beta1-integrin-mediated progenitor cell adhesion and migration are differentially regulated by external stimuli and suggest that this regulation corresponds to different activation states of beta1-integrins in hematopoietic progenitor cells. PMID- 9345037 TI - Mechanisms of stem cell factor and erythropoietin proliferative co-signaling in FDC2-ER cells. AB - Studies of hematopoietic progenitor cell development in vivo, ex vivo, and in factor-dependent cell lines have shown that c-kit promotes proliferation through synergistic effects with at least certain type 1 cytokine receptors, including the erythropoietin (Epo) receptor. Presently, c-kit is shown to efficiently support both mitogenesis and survival in the FDCP1 cell subline, FDC2. In this system, mitogenic synergy with c-kit was observed for ectopically expressed wild type Epo receptors (wt-ER), an epidermal growth factor (EGF) receptor/Epo receptor chimera, and a highly truncated Epo receptor construct ER-Bx1. Thus, the Epo receptor cytoplasmic box 1 subdomain appears, at least in part, to mediate mitogenic synergy with c-kit. In studies of potential effectors of this response, Jak2 tyrosine phosphorylation was shown to be induced by Epo, but not by stem cell factor (SCF). In addition and in contrast to signaling in Mo7e and BM6 cell lines, in FDC2-ER cells SCF and Epo each were shown to rapidly activate Pim 1 gene expression. Recently, roles also have been suggested for the nuclear trans factor GATA-1 in regulating progenitor cell proliferation. In FDC2-ER cells, the ectopic expression of GATA-1 had no detectable effect on Epo inhibition of apoptosis. However, GATA-1 expression did result in a selective and marked inhibition in mitogenic responsiveness to SCF and to a decrease in c-kit transcript expression. These studies of SCF and Epo signaling in FDC2-wt-ER cells serve to functionally map the ERB1 region as a c-kit-interactive domain, suggest that Pim1 might contribute to SCF and Epo mitogenic synergy and support the notion that SCF and Epo may act in opposing ways during red cell differentiation. PMID- 9345038 TI - Sensitization of hematopoietic stem and progenitor cells to trimetrexate using nucleoside transport inhibitors. AB - Antifolates such as methotrexate (MTX) and trimetrexate (TMTX) are widely used in the treatment of cancer and nonmalignant disorders. Transient, yet sometimes severe myelosuppression is an important limitation to the use of these drugs. It has previously been shown that clonogenic myeloid progenitors and colony-forming units-spleen are resistant to antifolates, suggesting that myelotoxicity occurs late in hematopoietic development. The goal of this study was to define the mechanisms by which primitive hematopoietic cells resist the toxic effects of antifolate drugs. To test the hypothesis that myeloid progenitors may salvage extracellular nucleotide precursors to resist TMTX toxicity, a defined liquid culture system was developed to measure TMTX toxicity in expanding progenitor populations. These in vitro experiments showed that both human and murine progenitors can resist TMTX toxicity by importing thymidine and hypoxanthine from the serum. As predicted from these findings, several drugs that block thymidine transport sensitized progenitors to TMTX in vitro, although to differing degrees. These nucleoside transport inhibitors were used to test whether progenitors and hematopoietic stem cells (HSCs) could be sensitized to TMTX in vivo. Treatment of mice with TMTX and nitrobenzylmercaptopurineriboside phosphate (NBMPR-P), a potent transport inhibitor, caused significant depletions of clonogenic progenitors within the bone marrow (20-fold) and spleen (6-fold). Furthermore, NBMPR-P administration dramatically sensitized HSCs to TMTX, with dual-treated mice showing a greater than 90% reduction in bone marrow repopulating activity. These studies demonstrate that both myeloid progenitor cells and HSCs resist TMTX by using nucleotide salvage mechanisms and that these pathways can be pharmacologically blocked in vivo using nucleoside transport inhibitors. These results have important implications regarding the use of transport inhibitors for cancer therapy and for using variants of dihydrofolate reductase for in vivo selection of genetically modified HSCs. PMID- 9345039 TI - In vivo characterization of recombinant von Willebrand factor in dogs with von Willebrand disease. AB - Hereditary von Willebrand factor (vWF ) deficiency in Dutch Kooiker dogs, which have undetectable levels of vWF, causes spontaneous hemorrhage of mucosal surfaces similar to the clinical picture of von Willebrand disease in humans. Therefore, we used this canine model to study the in vivo effects of a new recombinant von Willebrand factor (rvWF ) preparation containing all species of vWF multimers compared with a rvWF fraction containing only low molecular weight multimers (LMW-rvWF ) and with a plasma-derived factor VIII/vWF concentrate (pdvWF ). In the vWF-deficient dogs, the half-life of vWF:Ag was 21.6 and 22.1 hours for rvWF, 7.7 hours for pdvWF, and 9 hours for LMW-rvWF; in vivo recovery of vWF:Ag was 59%, 64%, and 70% for rvWF, 33% for pdvWF and 92% for LMW-rvWF; in vivo recovery of RCoF was 78%, 110%, and 120% for rvWF, and 25% for pdvWF. Both rvWF and pdvWF caused increases in factor VIII, which were sustained even when vWF:Ag had decreased to nearly undetectable levels and only monomeric or dimeric species were detectable on agarose gels. At the dosages used, no effect was seen on bleeding time, but the rate of blood flow from cuticle wounds was reduced after a single bolus administration of rvWF. The rvWF was able to control a severe nose bleed in one dog. PMID- 9345040 TI - Association of tissue factor pathway inhibitor with human umbilical vein endothelial cells. AB - Tissue factor pathway inhibitor (TFPI) is a serine protease inhibitor of the extrinsic coagulation system, synthesized in endothelial cells, which has recently been shown to play an important role in the regulation of activated coagulation factors at the endothelial cell surface. In the present study we investigated the subcellular localization and metabolism of TFPI in human umbilical vein endothelial cells (HUVEC). Immunocytochemical labeling of HUVEC with anti-TFPI showed specific labeling associated with the cell surface and with many intracellular organelles including the Golgi complex. Further characterization of these organelles was performed by colocalizing the anti-TFPI with 3-(2, 4-dinitroanilino)'-amino-N-methyldipropylamine (DAMP; to demonstrate low pH), mannose phosphate receptor (endosomes), and LAMP 1 (late endocytic compartments). TFPI also colocalized with antibodies to the human transferrin receptor, a marker for early endocytic, recycling compartment. Endogenous TFPI colocalized with biotin in intracellular vesicles during endocytosis after biotinylation of the cell surface, which indicated that TFPI was being co internalized with the surface biotin. The binding of exogenously added 125I-TFPI increased linearly to HUVEC over the concentration range of 0 to 32 nmol/L without saturation, the binding was not affected by up to a thousand-fold molar excess of unlabeled TFPI, and heparin inhibited the binding dose dependently. An intact C-terminal domain was important for the interaction between TFPI and the cell surface of HUVEC, because less than 10% of a C-terminal truncated form of TFPI (TFPI1-161) was bound after addition of equimolar concentrations of full length TFPI. Exogenously added 125I-TFPI was not degraded in HUVEC during 4 hours at 37 degrees C. The presence of TFPI in endocytic and recycling compartments support the hypothesis that endogenous, membrane-anchored TFPI could be internalized for subsequent recycling back to the cell surface. PMID- 9345041 TI - Thrombin stimulation of platelets induces plasminogen activation mediated by endogenous urokinase-type plasminogen activator. AB - Gene knockout mice studies indicate that urokinase-type plasminogen activator (u PA) is importantly involved in fibrinolysis, but its physiologic mechanism of action remains poorly understood. We postulated that platelets may be involved in this mechanism, as they carry a novel receptor for u-PA and a portion of the single-chain u-PA (scu-PA) intrinsic to blood is tightly associated with platelets. Therefore, plasminogen activation by platelet-associated u-PA was studied. When washed platelets were incubated with plasminogen, no plasmin was generated as detected by plasmin synthetic substrate (S2403) hydrolysis; however, after the addition of thrombin, but not other agonists, platelet-dependent plasminogen activation occurred. Plasminogen activation was surface-related, being inhibited by blocking platelet fibrinogen receptors or by preventing plasminogen binding to the thrombin-activated platelet surface. U-PA was identified as the only plasminogen activator responsible and enrichment of platelets with exogenous scu-PA significantly augmented plasminogen activation. These findings appeared paradoxical because thrombin inactivates scu-PA. Indeed, zymograms showed inactivation of scu-PA during the first hour of incubation with even the lowest dose of thrombin used (1 u/mL). However, this was followed by a thrombin dose-dependent (1 to 10 u/mL) partial return of u-PA activity. Reactivation of u-PA was not due to the direct action of thrombin, but required platelets and was found to be related to a platelet lysosomal thiol protease, consistent with cathepsin C. In conclusion, a new pathway of plasminogen activation by platelet-associated endogenous or exogenous scu-PA was demonstrated, which is specifically triggered by thrombin activation of platelets. These findings may help explain u-PA-mediated physiological fibrinolysis and have implications for therapeutic thrombolysis with scu-PA. PMID- 9345042 TI - Anti-CD43 inhibits monocyte-endothelial adhesion in inflammation and atherogenesis. AB - Recruitment of blood monocytes into tissues is a central event in the inflammatory response and in atherogenesis. The mechanisms leading to monocyte adhesion and migration through endothelium are not completely defined. We recently reported that MAb L11, against the leukocyte sialomucin CD43, blocks T lymphocyte binding to lymph node and Peyer's patch high endothelial venules (HEV) and inhibits T-cell extravasation from the blood into organized secondary lymphoid tissues. We have now assessed the ability of L11 to inhibit monocyte endothelial (EC) interactions and trafficking. L11 blocks binding of WEHI78/24 cells, a murine monocytoid cell line, to inflamed lymph node HEV and inhibits recruitment of monocytes and neutrophils to thioglycollate-inflamed peritoneum. Because monocyte adhesion to the endothelium and diapedesis in lesion-prone regions of the vasculature is among the earliest events in atherogenesis, leading to formation of lipid-laden foam cells, the ability of L11 to block monocyte recognition of aortic endothelial cells was assessed in a novel ex vivo assay of monocyte binding to intact rabbit aortic endothelium. Cholesterol feeding of rabbits induces enhanced aortic adhesiveness for monocytes and WEHI78/24 monocytoid cells, and this adhesion is inhibited by L11. The inhibitory effect of L11 is additive with that of a cocktail of anti-L-selectin and anti-alpha4 and beta2 integrin monoclonal antibodies. Thus, CD43 represents a novel target for manipulation of monocyte recruitment in inflammation and atherogenesis. PMID- 9345043 TI - Fibrin regulation of interleukin-8 gene expression in human vascular endothelial cells. AB - Recent studies in our laboratory, as well as others, have suggested that fibrin can regulate cell function in vitro and likely control inflammation in vivo by acting as a potent cell activator. This has led us to hypothesize that during tissue and vascular injury, fibrin can enhance leukocyte recruitment by inducing vascular endothelial cell expression of leukocyte chemotactic factors. To begin to test this hypothesis, we developed an in vitro model of in situ fibrin polymerization on human umbilical vein endothelial cell culture (HUVEC) and determined the ability of fibrin to induce HUVEC expression of the potent leukocyte chemotactic factor interleukin-8 (IL-8). Our initial studies showed that fibrin induced IL-8 expression in a time- and dose-dependent fashion. Fibrin induced IL-8 expression in HUVEC could be seen as early as 2 hours post-fibrin stimulation. Additionally, fibrin concentrations as low as 30 microg/mL stimulated a detectable level of IL-8 antigen expression from HUVEC. We also showed that this fibrin induced IL-8 had the identical molecular weight and similar antigenic identity as recombinant and monocyte derived IL-8. Northern blot analysis showed that the IL-8 antigen increase seen in fibrin treated HUVEC was due to fibrin induced elevation of steady state mRNA expression in HUVEC. These data clearly support our hypothesis that fibrin is a potent vascular endothelial cell (VEC) activator that can directly contribute to leukocyte recruitment and activation by inducing leukocyte chemotactic factor expression from VEC. PMID- 9345044 TI - Increased enzymatic activity of the T-cell antigen receptor-associated fyn protein tyrosine kinase in asymptomatic patients infected with the human immunodeficiency virus. AB - The immune system of patients infected with human immunodeficiency virus (HIV) is in a state of chronic activation; however, the nature of HIV-related immune activation is unknown. As normal T-cell activation involves early tyrosine phosphorylation induced by the T-cell antigen receptor-associated src-family protein tyrosine kinase p59(fyn(T)) (Fyn), we examined a potential role for this kinase in HIV-related immune dysfunction. We determined the relative specific kinase activity of Fyn in lysates of peripheral blood mononuclear cells from 47 normal control individuals tested negative for HIV-1 and -2, human T-cell lymphotropic virus Type I, hepatitis B virus (HBV), hepatitis C virus (HCV), and syphilis; 14 asymptomatic HIV-infected patients having near-normal CD4+ T-cell counts (350 to 980 CD4+ cells/microL); 4 patients with symptomatic acquired immunodeficiency syndrome (AIDS) (<30 CD4+ cells/microL); 13 patients having chronic infection with HBV (6 patients) or HCV (7 patients); and 6 patients with systemic lupus erythematosis (SLE). All patients with asymptomatic HIV disease were shown to have a profound increase (mean increase of 19-fold; range threefold to 56-fold increase; p = 1.33 x 10(-9)) in the relative specific kinase activity of Fyn compared to uninfected controls or patients with hepatitis or SLE. In contrast, patients with AIDS had an Fyn-specific kinase activity that was much less affected (mean increase of threefold; range onefold to sevenfold increase; p = 1.30 x 10(-5)). It was further shown that HIV infection affects the Fyn specific kinase activity in CD8+-enriched cells, suggesting abnormal Fyn activity in both CD8+ as well as CD4+ T lymphocytes. Initial results implicate a role for the CSK protein tyrosine kinase as responsible for the abnormal Fyn kinase activity observed in HIV-infected patients. These data indicate early and chronic activation of Fyn as a unique HIV-related effect that has the potential to be diagnostic for early HIV infection and/or may serve as a prognostic indicator for advancement to full-blown AIDS. More importantly, sustained activation of the protein tyrosine kinase associated with T-cell antigen receptor function may result in, or contribute to, the immunopathogenic effects associated with HIV infection. PMID- 9345045 TI - Regulated expression of the Eph-related receptor tyrosine kinase Hek11 in early human B lymphopoiesis. AB - Members of the large Eph family of receptor tyrosine kinases (RTKs) display temporally and spatially restricted expression patterns during embryogenesis, suggesting a role in various developmental processes. We have begun to investigate the expression of members of this receptor family during human hematopoiesis, in particular B lymphopoiesis. Expression of Eph RTKs in cells of the B-lymphoid lineage was assessed by using degenerate oligonucleotide primers based on stretches of conserved nucleic acid sequences in members of the Eph family. First, the content of Eph-family RTKs was assessed in freshly sorted fetal bone marrow pro-B cells. This population was found to harbor transcripts of the Hek8 and Hek11 members of this gene family. Subsequent analysis of expression of these genes in B cells representing various differentiation and ontogenic stages showed that the Hek8 transcript is constitutively present in all fetal and adult B-lineage cells, with high levels of expression in peripheral blood B cells. In contrast, the Hek11 transcript was exclusively found in fetal bone marrow pro-B cells and pre-B cells, but not in more mature fetal B-lineage cells. All adult B-lineage cells, from early pro-B cells to end-stage plasma cells, lacked Hek11 transcripts. The developmentally regulated expression of Hek11 during fetal B lymphopoiesis suggests a role for this gene in pre/pro-B cell expansion and/or differentiation and defines a difference in progenitor B cell populations isolated from fetal versus adult human bone marrow. PMID- 9345046 TI - Molecular mimicry between the rabies virus glycoprotein and human immunodeficiency virus-1 GP120: cross-reacting antibodies induced by rabies vaccination. AB - The 160-170 sequence of human immunodeficiency virus (HIV)-1 gp120 mimics a nicotinic receptor-binding motif of rabies virus glycoprotein and snake neurotoxins. This sequence has been proposed to be involved in the binding of HIV 1 gp120 to the acetylcholine binding sites of nicotinic receptors. By using biomolecular interaction analysis (BIA) technology we have found that HIV-1 gp120 can bind to detergent-extracted nicotinic receptor from fetal calf muscle. The binding is inhibited by nicotine and by a synthetic peptide reproducing the gp120 160-170 sequence. The molecular mimicry between gp120 and rabies virus glycoprotein is confirmed by cross-reacting antibodies. We have found that vaccination against rabies can induce the production of anti-HIV-1 gp120 antibodies in humans. The cross-reacting antibodies are directed to the gp120 sequence involved in the mimicry with the rabies virus glycoprotein. The cross reactivity between the rabies virus and HIV-1 has important implications in transfusion medicine. Moreover, the presence of cross-reacting antibodies between the nicotinic receptor binding site of rabies virus glycoprotein and a fragment of HIV-1 gp120 strengthens the hypothesis about the possible role of nicotinic receptors as potential receptors for HIV-1 in the central nervous system. PMID- 9345047 TI - Fas-independent apoptosis of activated T cells induced by antibodies to the HLA class I alpha1 domain. AB - In addition to their major function in antigen presentation and natural killer cell activity regulation, HLA class I molecules may modulate T-cell activation and proliferation. Monoclonal antibodies (MoAbs) that recognize distinct epitopes of HLA class I molecules were reported to interfere with T-cell proliferation. We show here that two MoAbs (mouse MoAb90 and rat YTH862) that bind to an epitope of the alpha1 domain of HLA class I heavy chain induce apoptotic cell death of activated, but not resting, peripheral T lymphocytes. Other reference anti-HLA class I antibodies specific for distinct epitopes of the alpha1 (B9.12.1), alpha2 (W6/32), or alpha3 (TP25.99) domains of the heavy chain decreased T-cell proliferation but had little or no apoptotic effect. Apoptosis shown by DNA fragmentation, phosphatidylserine externalization, and decrease of mitochondrial transmembrane potential was observed whatever the type of T-cell activator. Apoptosis did not result from Fas/Fas-L interaction and distinct though partly overlapping populations of activated T cells were susceptible to Fas- and HLA class I-mediated apoptosis, respectively. Induction of apoptosis did not require HLA class I cross-linking inasmuch as it could be observed with monovalent Fab' fragments. The data indicate that MoAb90 and YTH862 directed against the alpha1 domain of HLA class I trigger apoptosis of activated T lymphocytes by a pathway which does not involve Fas-ligand. PMID- 9345048 TI - A monocyte conditioned medium is more effective than defined cytokines in mediating the terminal maturation of human dendritic cells. AB - Mature human dendritic cells can be generated in substantial numbers from nonproliferating progenitors in human blood using a two-step protocol. T cell depleted mononuclear cells are first cultured with granulocyte-macrophage colony stimulating factor and interleukin-4 (IL-4) and then exposed to monocyte conditioned medium (MCM). The dendritic cells generated using this approach are rendered terminally mature and are the most potent antigen presenting cells identified to date in humans. We sought to characterize factors in MCM that induce the terminal differentiation of dendritic cells. MCM contained substantial, although varying, quantities of several factors including tumor necrosis factor-alpha, IL-1beta, IL-6, and interferon-alpha. However, none of the four factors, individually or in various combinations, could fully substitute for the MCM to generate irreversibly differentiated dendritic cells. The yields, percentage of cells expressing the mature phase marker CD83, and mixed leukocyte reaction-stimulatory function were lower when defined cytokines were used in the place of MCM. Therefore, the full maturation of dendritic cells, because it entails changes in many known cell and molecular properties, requires a number of different cytokines that are released in tandem from appropriately stimulated monocytes. We propose that MCM-matured dendritic cells will be the most effective adjuvants for immunotherapy in vivo. PMID- 9345049 TI - Stem cell factor enhances interleukin-2-mediated expansion of murine natural killer cells in vivo. AB - The administration of low dose interleukin-2 (IL-2) results in a selective expansion of natural killer (NK) cells in vivo, and promotes the differentiation of NK cells from hematopoietic precursor cells in vitro. We have previously shown that stem cell factor (SCF ), the ligand to the c-kit tyrosine kinase receptor, enhances IL-2-induced NK cell proliferation and differentiation in vitro. Here, we investigated the effects of SCF plus IL-2 delivered to mice in vivo. Eight week-old C57BL/6 mice were treated with a continuous subcutaneous infusion of IL 2 (1 x 10(4) IU/d) plus a daily intraperitoneal dose of SCF (100 microg/kg/d), IL 2 alone, SCF alone, or vehicle alone for 8 weeks. The in vivo serum concentration of IL-2 ranged between 352 +/- 12.0 pg/mL and 606 +/- 9.0 pg/mL, achieving selective saturation of the high affinity IL-2 receptor, while the peak SCF serum concentration was 296 +/- 13.09 ng/mL. Alone, the daily administration of SCF had no effect on the expansion of NK cells. The continuous infusion of IL-2 alone did result in a significant expansion of NK1.1+CD3- cells compared to mice treated with placebo or SCF. However, mice treated with both SCF and IL-2 showed an increase in the absolute number of NK cells that was more than twofold that seen with IL-2 alone, in the spleen (P A mutation was found 11 bp upstream of exon 3. In our screening of 102 chromosomes from unrelated individuals, this base-pair substitution was not found, indicating that it was not a polymorphism. mRNA analysis revealed that splicing involved a cryptic splice site, located 71/70 bp upstream of exon 3, resulting in generation of mRNA with an insert of 69 nucleotides. In addition, a small amount of a transcript with a deleted exon 3 and a very low level of wild type transcript were detected. Translation of the extended transcript resulted in an androgen-receptor protein with 23 amino acid residues inserted between the two zinc clusters, displaying defective DNA binding and defective transcription activation. PMID- 9345100 TI - CFTR gene mutations in men with bilateral ejaculatory-duct obstruction and anomalies of the seminal vesicles. PMID- 9345101 TI - The role of MMAC1 mutations in early-onset breast cancer: causative in association with Cowden syndrome and excluded in BRCA1-negative cases. AB - Cowden syndrome (CS) is an autosomal dominant disorder associated with the development of hamartomas and benign tumors in a variety of tissues, including the skin, thyroid, breast, endometrium, and brain. It has been suggested that women with CS are at increased risk for breast cancer. A locus for CS was recently defined on chromosome 10 in 12 families, resulting in the identification of the CS critical interval, between the markers D10S215 and D10S541. More recently, affected individuals in four families with CS have been shown to have germ-line mutations in a gene known as "PTEN," or "MMAC1," which is located in the CS critical interval on chromosome 10. In this study, we report three novel MMAC1 mutations in CS and demonstrate that MMAC1 mutations are associated with CS and breast cancer. Furthermore, we also show that certain families and individuals with CS do not have mutations in the coding sequence of MMAC1. Finally, we did not detect MMAC1 mutations in a subpopulation of individuals with early-onset breast cancer, suggesting that germ-line mutations in this gene do not appear to be common in this group. PMID- 9345102 TI - De novo rearrangements found in 2% of index patients with spinal muscular atrophy: mutational mechanisms, parental origin, mutation rate, and implications for genetic counseling. AB - Spinal muscular atrophy (SMA) is a relatively common autosomal recessive neuromuscular disorder. We have identified de novo rearrangements in 7 (approximately 2%) index patients from 340 informative SMA families. In each, the rearrangements resulted in the absence of the telomeric copy of the survival motor neuron (SMN) gene (telSMN), in two cases accompanied by the loss of the neuronal apoptosis-inhibitory protein gene . Haplotype analysis revealed unequal recombination in four cases, with loss of markers Ag1-CA and C212, which are near the 5' ends of the SMN genes. In one case, an interchromosomal rearrangement involving both the SMN genes and a regrouping of Ag1-CA and C212 alleles must have occurred, suggesting either interchromosomal gene conversion or double recombination. In two cases, no such rearrangement was observed, but loss of telSMN plus Ag1-CA and C212 alleles in one case suggested intrachromosomal deletion or gene conversion. In six of the seven cases, the de novo rearrangement had occurred during paternal meiosis. Direct detection of de novo SMA mutations by molecular genetic means has allowed us to estimate for the first time the mutation rate for a recessive disorder in humans. The sex-averaged rate of 1.1 x 10(-4), arrived at in a proband-based approach, compares well with the rate of 0.9 x 10(-4) expected under a mutation-selection equilibrium for SMA. These findings have important implications for genetic counseling and prenatal diagnosis in that they emphasize the relevance of indirect genotype analysis in combination with direct SMN-gene deletion testing in SMA families. PMID- 9345103 TI - Localization of the congenital dyserythropoietic anemia II locus to chromosome 20q11.2 by genomewide search. AB - Congenital dyserythropoietic anemias (CDA) are genetic disorders characterized by anemia and ineffective erythropoiesis. Three main types of CDA have been distinguished: CDA I and CDA III, whose loci have been already mapped, and CDA II (MIM 224100), the most frequent among CDAs, which is transmitted as an autosomal recessive trait and is known also as "HEMPAS" (hereditary erythroblast multinuclearity with positive acidified serum). We have recruited a panel of well characterized CDA II families and have used them to search for the CDA II gene by linkage analysis. After the exclusion of three candidate genes, we ob-tained conclusive evidence for linkage of CDA II to microsatellite markers on the long arm of chromosome 20 (20q11.2). A maximum two-point LOD score of 5.4 at a recombination fraction of .00 was obtained with marker D20S863. Strong evidence of allelic association with the disease was detected with the same marker. Some recombinational events established a maximum candidate interval of approximately 5 cM. PMID- 9345105 TI - Evidence for locus heterogeneity in Puerto Ricans with Hermansky-Pudlak syndrome. AB - Hermansky-Pudlak syndrome (HPS) consists of ocu-locutaneous albinism, a platelet storage-pool deficiency, and ceroid lipofuscinosis. In a recent report on the cloning of an HPS gene, all 22 Puerto Rican HPS patients were homozygous for a 16 bp duplication in exon 15. This presumably reflected a founder effect for the HPS mutation in Puerto Rico. Nevertheless, we ascertained two individuals from central Puerto Rico who lacked the 16-bp duplication, exhibited significant amounts of normal-size HPS mRNA by northern blot analysis, and had haplotypes in the HPS region that were different from the haplotype of every 16-bp-duplication patient. Moreover, these two individuals displayed no mutations in their cDNA sequences, throughout the entire HPS gene. Both patients exhibited pigment dilution, impaired visual acuity, nystagmus, a bleeding diathesis, and absent platelet dense bodies, confirming the diagnosis of HPS. These findings indicate that analysis of Puerto Rican patients for the 16-bp duplication in HPS cannot exclude the diagnosis of HPS. In addition, HPS most likely displays locus heterogeneity, consistent with the existence of several mouse strains manifesting both pigment dilution and a platelet storage-pool deficiency. PMID- 9345104 TI - Familial nontoxic multinodular thyroid goiter locus maps to chromosome 14q but does not account for familial nonmedullary thyroid cancer. AB - Thyroid goiter is a common condition that is often associated with iodine deficiency. Familial forms of goiter in areas not known to feature iodine deficiency are much less common. We have performed a genomic search on a single large Canadian family with 18 cases of nontoxic multinodular goiter in which 2 individuals also had papillary lesions highly suggestive of papillary carcinoma. A locus on chromosome 14q (MNG1 [multinodular goiter 1]) has been identified, with a maximal two-point LOD score of 3.8 at D14S1030 and a multipoint LOD score of 4.88 at the same marker, defined by D14S1062 (upper boundary) and D14S267 (lower boundary). The gene encoding thyroid-stimulating hormone receptor (TSHR), which is located on chromosome 14q, is outside the linked region. To determine the role of this gene in familial nonmedullary thyroid cancer (NMTC), we studied 37 smaller pedigrees each containing at least two cases of NMTC. Analysis by both parametric and nonparametric methods indicates that only a very small proportion of familial NMTC (point estimate 0.001, support intervals 0-.6 under a dominant model) is attributable to MNG1. PMID- 9345106 TI - Mutations in the TIGR gene in familial primary open-angle glaucoma in Japan. PMID- 9345107 TI - Progressive ataxia due to a missense mutation in a calcium-channel gene. AB - We describe a family with severe progressive cerebellar ataxia involving the trunk, the extremities, and speech. The proband, who has prominent atrophy of the cerebellum, shown by magnetic resonance imaging, was confined to a wheelchair at the age of 44 years. Two sons have episodes of vertigo and ataxia that are not responsive to acetazolamide. Quantitative eye-movement testing showed a consistent pattern of abnormalities localizing to the cerebellum. Genotyping suggested linkage to chromosome 19p, and SSCP showed an aberrant migrating fragment in exon 6 of the calcium-channel gene CACNA1A, which cosegregated with the disease. Sequencing of exon 6 identified a G-->A transposition in one allele, at nucleotide 1152, resulting in a predicted glycine-to-arginine substitution at codon 293. The CAG-repeat expansion associated with spinocerebellar ataxia 6 was not present in any family members. This family is unique in having a non-CAG repeat mutation that leads to severe progressive ataxia. Since a great deal is known about the function of calcium channels, we speculate on how this missense mutation leads to the combination of clinical symptoms and signs. PMID- 9345109 TI - Childhood cancer and neural tube defects. PMID- 9345108 TI - Evidence that the penetrance of mutations at the RP11 locus causing dominant retinitis pigmentosa is influenced by a gene linked to the homologous RP11 allele. AB - A subset of families with autosomal dominant retinitis pigmentosa (RP) display reduced penetrance with some asymptomatic gene carriers showing no retinal abnormalities by ophthalmic examination or by electroretinography. Here we describe a study of three families with reduced-penetrance RP. In all three families the disease gene appears to be linked to chromosome 19q13.4, the region containing the RP11 locus, as defined by previously reported linkage studies based on five other reduced-penetrance families. Meiotic recombinants in one of the newly identified RP11 families and in two of the previously reported families serve to restrict the disease locus to a 6-cM region bounded by markers D19S572 and D19S926. We also compared the disease status of RP11 carriers with the segregation of microsatellite alleles within 19q13.4 from the noncarrier parents in the newly reported and the previously reported families. The results support the hypothesis that wild-type alleles at the RP11 locus or at a closely linked locus inherited from the noncarrier parents are a major factor influencing the penetrance of pathogenic alleles at this locus. PMID- 9345110 TI - Loss of heterozygosity or: how I learned to stop worrying and love mitotic recombination. PMID- 9345111 TI - "Mistakes happen": somatic mutation and disease. PMID- 9345112 TI - Understanding human cancer in a fly? PMID- 9345113 TI - DNA variation and language affinities. PMID- 9345114 TI - Longitudinal relation between endogenous testosterone and cardiovascular disease risk factors in middle-aged men. A 13-year follow-up of former Multiple Risk Factor Intervention Trial participants. AB - The present study examined lifestyle and behavioral correlates of the change in total testosterone over 13 years in 66 men aged 41-61 years who were former participants of the Multiple Risk Factor Intervention Trial (MRFIT) at the Pittsburgh, Pennsylvania, center. The authors also determined in these men if changes in total testosterone are related to changes in cardiovascular disease risk factors. The mean total testosterone level was 751 (standard deviation, 248) ng/dl at baseline and decreased by 41 (standard deviation, 314) ng/dl during follow-up. The correlation between measures was r = 0.44 (p < 0.001). In multivariate analysis, higher type A coronary-prone behavior score, greater pack years of cigarette smoking, and the MRFIT special intervention group were associated with larger decreases in total testosterone. Age, body weight, weight change, leisure time activity level, and alcohol intake were not related to the change in total testosterone. The decrease in endogenous testosterone was associated with an increase in triglycerides and a decrease in high density lipoprotein cholesterol in multivariate analysis controlling for obesity and other lifestyle covariates. There was little relation between change in testosterone and change in total and low density lipoprotein cholesterol or blood pressure. This longitudinal study confirms a gradual decline in total testosterone levels with advancing age in older men and provides evidence that lifestyle and psychosocial factors are related to this decline. Decreases in endogenous testosterone levels with age in men are associated with potentially unfavorable changes in triglycerides and high density lipoprotein cholesterol. PMID- 9345116 TI - Risk factors for hospitalization in a cohort with type 1 diabetes. Wisconsin Diabetes Registry. AB - The authors investigate the postonset hospitalization rate and risk factors during 1987-1994 in Wisconsin, in a population-based, incidence cohort followed from diagnosis of Type 1 diabetes mellitus at ages 0-29 (n = 577). The overall rate was 8.9 +/- 0.60 (standard error) per 100 person-years of diabetes, whereof 5.7 was due to hyperglycemia, 1.9 to hypoglycemia, and 1.3 to other and undetermined causes. Major risk factors for hospitalization were longitudinally measured glycosylated hemoglobin level (rate ratio = 1.5 per 2% increase, 95% confidence interval 1.4-1.7), black/other race (rate ratio = 1.9, 95% confidence interval 1.0-3.6), diagnosis in a non-university-based setting (rate ratio = 1.9, 95% confidence interval 1.2-3.2), female sex (rate ratio = 1.5, 95% confidence interval 1.0-2.4 at age 11), age in males (rate ratio = 0.6, 95% confidence interval 0.4-0.8 per 5-year increase), and public or no insurance up to 18 months postdiagnosis (rate ratio = 2.2, 95% confidence interval 1.1-4.4). For individuals less than 18 years, "black/other race" was replaced in the model by "having other than two biologic parents in the home" (rate ratio = 2.0, 95% confidence interval 1.1-3.5). Hence, hospitalization is common in children, adolescents, and young adults with diabetes, primarily for problems with glycemic control. PMID- 9345115 TI - Lycopene and myocardial infarction risk in the EURAMIC Study. AB - A multicenter case-control study was conducted to evaluate the relations between antioxidant status assessed by biomarkers and acute myocardial infarction. Incidence cases and frequency matched controls were recruited from 10 European countries to maximize the variance in exposure within the study. Adipose tissue needle aspiration biopsies were taken shortly after the infarction and analyzed for levels of carotenoids and tocopherols. An examination of colinearity including all covariates and the three carotenoids, alpha-carotene, beta carotene, and lycopene, showed that the variables were sufficiently independent to model simultaneously. When examined singularly, each of the carotenoids appeared to be protective. Upon simultaneous analyses of the carotenoids, however, using conditional logistic regression models that controlled for age, body mass index, socioeconomic status, smoking, hypertension, and maternal and paternal history of disease, lycopene remained independently protective, with an odds ratio of 0.52 for the contrast of the 10th and 90th percentiles (95% confidence interval 0.33-0.82, p = 0.005). The associations for alpha- and beta carotene were largely eliminated. We conclude that lycopene, or some substance highly correlated which is in a common food source, may contribute to the protective effect of vegetable consumption on myocardial infarction risk. PMID- 9345117 TI - Epidemiology of acute low back injury in employees of a large home improvement retail company. AB - Acute low back injuries are described in a cohort of about 31,000 material handlers employed in all Home Depot, Inc., retail stores in California from 1990 through 1994. With over 87 million work hours, incidence density rates, rate ratios, and confidence intervals are given by age, sex, length of employment, and job-lifting requirements. Injuries are further described by lost work days, activity at time of injury, work restrictions, and time frames. The unadjusted low back injury rate per million work hours was 1.6 times higher for men compared with women, and rates were highest for those less than 25 years of age, those with less than 2 years of current job experience, and employees with the greatest materials lifting and handling job requirements. These findings in unadjusted rates and rate ratios persisted when each was adjusted through a Poisson regression model, with the exception of sex. The adjusted risk ratio for males was reversed with significantly higher risk in females when the rate ratio was adjusted for age, lifting intensity, and length of job experience. Injuries were most commonly associated with lifting activities and, while injury occurrence was highest from 10 a.m. to 4 p.m., rates were greatest during those hours when the store was closed to retail activities. Merchandise stocking that requires heavy and frequent materials handling is done during these hours. Fewer injuries than expected were reported on weekends, days with considerably less materials handling activities. PMID- 9345118 TI - Measles incidence, case fatality, and delayed mortality in children with or without vitamin A supplementation in rural Ghana. AB - Data on measles incidence, acute case fatality, and delayed mortality were collected on 25,443 children aged 0-95 months during the course of a community based, double-blind, placebo-controlled, randomized trial of vitamin A supplementation in rural, northern Ghana between 1989 and 1991. Measles vaccine coverage in these children was 48%. The overall estimated measles incidence rate was 24.3 per 1,000 child-years, and acute case fatality was 15.7%. There was not significantly increased mortality in survivors of the acute phase of measles compared with controls (rate ratio = 1.22, 95% confidence interval (CI) 0.65 2.30). Reported incidence rates and case fatality were higher in families with low paternal education, in the dry season, and in unvaccinated children, and case fatality was higher in malnourished children. There was no sex difference in incidence, but acute case fatality was somewhat higher in girls than boys (adjusted odds ratio = 1.3, 95% CI 0.9-2.1). Measles incidence was lower in vitamin A-supplemented groups (23.6 per 1,000 child-years) than in placebo groups (28.9 per 1,000 child-years), but this difference was not statistically significant (p = 0.33). Among 946 measles cases in clusters randomized to receive vitamin A or placebo, there was no marked difference in acute measles case fatality between vitamin A-supplemented and placebo groups (15.4% vs. 14.5%, respectively). The biologic effects of vitamin A supplemented on the subsequent clinical manifestations and severity of measles need further elucidation. PMID- 9345119 TI - Role of viral load in heterosexual transmission of human immunodeficiency virus type 1 by blood transfusion recipients. Transfusion Safety Study Group. AB - Eighteen transfusion recipients infected with human immunodeficiency virus type 1 (HIV-1) were followed prospectively with their 19 long-term sexual partners from 1986 to 1993 in California, Florida, and New York. Follow-up included clinical, behavioral, immunologic, serologic, and virologic evaluations. Two partners were already infected when seen 18 and 34 months after sexual contact began following the infectious transfusion. Four of 17 initially seronegative partners seroconverted during 23 person-years of observation. The recipient's clinical status, mononuclear cell subset variations, and time trend in CD4+ counts had no association with transmission. Individual plasma HIV-1 ribonucleic acid (RNA) loads were stable during observation, and sexual transmission was not attributable to an upward trend or transient burst in viremia. However, recipients who transmitted HIV-1 to their sexual partners had higher mean viral RNA levels than did nontransmitting recipients (4.3 vs. 3.6 log10 copies/ml; p = 0.05). Although this series was small, the prospective observations suggest that viral load was the only characteristic in the recipient that contributed to heterosexual infectiousness. PMID- 9345120 TI - Estimation of human immunodeficiency virus (HIV) seroincidence among repeat anonymous testers in San Francisco. AB - The authors approximated human immunodeficiency virus (HIV) seroincidence in a population of men who have sex with men and who sought repeated anonymous HIV testing in San Francisco in 1995. The number of seroconversions and person-years of observation were estimated using the date and result of the current test and the self-reported date and result of the previous test. Estimates for HIV seroincidence (2.8 per 100 person-years, 95% confidence interval 2.3-3.4) and predictors of seroconversion were similar to those estimated from a prospective study of men who have sex with men conducted in San Francisco at the same time. While the limitations of self-reported data in a self-selected population are recognized, data from repeat testers may provide a practical surveillance tool. PMID- 9345121 TI - Vector diagnostics in dementia derived from Bayes' theorem. AB - This paper introduces the concept of vector diagnostics. In contrast to the conventional approach where one diagnosis takes precedence, the authors propose an alternative strategy that addresses the clinical reality of comorbidity and multiple diagnoses for an individual. Based on a Bayesian approach, the probability distribution for the etiologically heterogeneous dementia diagnoses is estimated from the Canadian Study of Health and Aging database. These data were collected between February 1991 and May 1992. This method facilitates the establishment of a probability for more than one diagnosis within a given individual. By analyzing the correspondence between diagnostic groups, it is demonstrated that some clinical diagnoses are not reliably distinguished on the basis of the considered subset of symptoms and signs. As a consequence, the conventional diagnostic categories might require revision. The resulting probabilistic algorithm allows for the mining of existing epidemiologic databases for patterns of signs and symptoms that characterize emerging diagnostic categories which might better account for the heterogeneity of the dementia subtypes and individual variability. PMID- 9345122 TI - Examination of "early mortality exclusion" as an approach to control for confounding by occult disease in epidemiologic studies of mortality risk factors. AB - Methods for the estimation of the effects of chronic disease risk factors on mortality continue to be an area that generates confusion and controversy. In response to the frequently observed U- or J-shaped relations between risk factors and mortality, some authors suggest that subjects dying during the first k years of follow-up (where k is some positive number less than the total length of follow-up) be excluded from statistical analyses. By excluded, the authors mean completely removed from the data set. The rationale is that persons dying during the first k years are likely to have a preexisting occult disease that confounds the relation between the risk factor under study and mortality. Excluding persons dying during the first k years of follow-up purportedly reduces this confounding. However, the authors are aware of no demonstration that this procedure effectively accomplishes its goal. They show that excluding subjects who die during the first k years of follow-up does not necessarily lead to a reduction in bias in the estimated effect of a risk factor on mortality when this relation is confounded by the presence of occult disease. Moreover, it is possible for such exclusion to exacerbate the confounding due to preexisting disease. Thus, excluding subjects dying during the first k years of follow-up is not necessarily an effective strategy for dealing with confounding due to occult disease. Investigators are encouraged to pursue alternative methods. PMID- 9345123 TI - Re: "Risk factors for non-Hodgkin's lymphomas in acquired immunodeficiency syndrome (AIDS)". PMID- 9345124 TI - Re: "Cancer incidence near radio and television transmitters in Great Britain". PMID- 9345125 TI - Re: "Serum transferrin saturation, stroke incidence, and mortality in women and men. The NHANES I Epidemiologic Followup Study". PMID- 9345144 TI - Not arguing, just asking about April case report. PMID- 9345145 TI - In favor of semirapid expansion. PMID- 9345143 TI - Comment about making outcome of treatment more predictable. PMID- 9345146 TI - The use of lingual appliances for correction of bimaxillary protrusion (four premolars extraction). AB - This is the case report of a 30-year-old actress who presented with a Class I bimaxillary protrusion. Because of esthetic concerns related to her public career, she was treated with a lingual appliance. An extraction of four premolars resulted in a greatly improved facial profile. PMID- 9345147 TI - Infraorbital abscess from orthodontic headgear. PMID- 9345148 TI - Treatment of a Class II malocclusion with impacted maxillary central incisors. PMID- 9345149 TI - Nonextraction treatment of a Class II, division 1 malocclusion with headgear and functional appliances. PMID- 9345150 TI - Three-dimensional effects in retraction appliance design. AB - The clinical importance of the three-dimensional effects of the force systems supplied by appliance designs used for retraction has long been appreciated. However, quantification of these force systems is not as well known. In this work, a numerical method is used to provide quantitative insight into three dimensional effects for typical appliance designs. One problem that occurs clinically is the axial rotation of a single rooted tooth as a result of the forces being applied by the retraction device on the tooth's buccal surface. An out-of-plane preactivated bend can be used to counteract this rotation. The proposed numerical method can accurately determine the force systems resulting from this out-of-plane preactivation as well as the in-plane force systems. It is shown that the out-of-plane effects are independent of the in-plane behaviour so that the usual forces and moment to force ratios are maintained. PMID- 9345151 TI - Continuous arch wire closing loop design, optimization, and verification. Part I. AB - In Part I, a systematic approach to closing loop design for use in continuous arch wires is presented. The design process uses Castigliano's theorem to derive equations for moment-to-force ratio (M/F) in terms of loop geometry. The equations are used to optimize designs by optimizing M/F to produce tooth movement via translation. Further refinements are performed with finite element simulations of designs. In Part II, predicted results are verified experimentally. The result of this process is a new design, the Opus loop, which is capable of delivering a nonvarying target M/F within the range of 8.0 to 9.1 mm inherently, without adding residual moments via twist or bends (commonly gable bends) anywhere in the arch wire or loop before insertion. The resulting precise force systems delivered with nonvarying M/F can move groups of teeth more accurately to achieve predetermined anteroposterior treatment goals for esthetics and/or stability. In Part II the experimental results show that the loops must be bent accurately to achieve their design potential. The negative impact on M/F of various dimensional changes to the loop design are presented. Experimental data are presented illustrating the improved performance of the new design over standard available designs. Suggested applications of the design for varying anchorage requirements are presented, along with a case report in which rigorous protraction requirements were met. PMID- 9345152 TI - Changes in jaw movement and jaw closing muscle activity after orthodontic correction of incisor crossbite. AB - The possible influences of the direction of occlusal loading delivered to the incisors in the sagittal direction during chewing on jaw movement and jaw closing muscle activity were investigated. Ten healthy children with crossbite of one or two incisors on the right side were selected. Each subject chewed a piece of chewing gum on the right side, and jaw displacements and electromyographic signals from the posterior temporalis and superficial masseter muscles on the ipsilateral side were sampled simultaneously. After orthodontic correction of the incisor crossbite relationship, identical records were taken. The inclinations of the gliding contacts for each posterior tooth in the lateral jaw excursion position were consistent before and after the treatment. The posttreatment records showed broader jaw movement patterns in the frontal view and faster jaw movement velocity in the lateral direction at a level close to the habitual maximum intercuspation position, when compared with the pretreatment records (P < 0.05). The duration of the muscle activity and the incidence of the silent periods of the masseter muscle during chewing significantly decreased after the treatment (P < 0.05). The current results give a neurophysiologic rationale for explaining the significance of orthodontic treatment in improving lowered masticatory efficiency in the way that the change in direction of the occlusal load achieved by tooth movement influences on the periodontal sensory input, which, in turn, modifies the trigeminal motor output and thus, eventually, jaw muscle activities. PMID- 9345153 TI - The functional matrix hypothesis revisited. 4. The epigenetic antithesis and the resolving synthesis. AB - In two interrelated articles, the current revision of the functional matrix hypothesis extends to a reconsideration of the relative roles of genomic and of epigenetic processes and mechanisms in the regulation (control, causation) of craniofacial growth and development. The dialectical method was chosen to analyze this matter, because it explicitly provides for the fuller presentation of a genomic thesis, an epigenetic antithesis, and a resolving synthesis. The later two are presented here, where the synthesis suggests that both genomic and epigenetic factors are necessary causes, that neither alone is also a sufficient cause, and that only the two, interacting together, furnish both the necessary and sufficient cause(s) of ontogenesis. This article also provides a comprehensive bibliography that introduces the several new, and still evolving, disciplines that may provide alternative viewpoints capable of resolving this continuing controversy; repetition of the present theoretical bases for the arguments on both sides of these questions seems nonproductive. In their place, it is suggested that the group of disciplines, broadly termed Complexity, would most likely amply repay deeper consideration and application in the study of ontogenesis. PMID- 9345154 TI - Orthodontic graduate education survey 1983-1994. PMID- 9345155 TI - An estimation of craniofacial growth in the untreated Class III female with anterior crossbite. AB - The literature has little to say regarding the normal growth and development of untreated individuals with Class III malocclusion or anterior crossbite. In part, this paucity of information is because of the relatively low prevalence of these characteristics in European-American populations and the need, recognized by the lay public and health professionals, for treatment of these conditions. Given the absence of true longitudinal data, this study attempts to estimate the growth of the untreated individual with Class III malocclusion and anterior crossbite by evaluating large samples of untreated subjects at distinct developmental stages. Initially the morphologic characteristics of 2074 Japanese female patients who had anterior crossbite were evaluated cephalometrically before treatment. On the basis of the cephalometric analysis, all subjects who did not have a Class III molar relationship were excluded from further analysis, leaving a sample of 1376. The subjects then were classified into seven groups (120-256 subjects per group) according to Hellman's stages of dental development. Descriptive statistics for 28 measurements were calculated. The results of this study imply that the maxilla in Japanese females maintains a retruded relationship to the cranial base and does not become less retrusive with time. In contrast, the mandible is protrusive even in the late deciduous dentition and becomes more protrusive with time, making the discrepancy between the upper and lower jaws progressively more severe. Dental compensations in both arches become increasingly evident as development progresses, and the underlying skeletal and dentoalveolar imbalances also are reflected in the soft tissue profile. PMID- 9345156 TI - Condyle displacement associated with premolar extraction and nonextraction orthodontic treatment of Class I malocclusion. AB - This study assessed condyle position change with premolar extraction and nonextraction orthodontic treatment in Class I malocclusions. Axially corrected pretreatment and posttreatment tomograms were obtained in 22 extraction and 13 nonextraction cases. Tomographic images were randomized and blinded for joint space measurement. A total of 27 linear anterior, superior, and posterior joint spaces were obtained from each tomogram and averaged. Comparisons of pretreatment and posttreatment joint spaces between groups were done by t test (pool variance estimate) with p < 0.05. Left and right anterior joint spaces were significantly increased during orthodontic treatment of the nonextraction group. No other significant changes in condyle position were determined in either group. There were no significant correlations between mean joint space changes with length of Class II elastic wear. There was no significant difference in condyle position change with extraction space closure using closing arch wires compared with elastic chain. PMID- 9345157 TI - Efficacy of intraarch mechanics using differential moments for achieving anchorage control in extraction cases. AB - A prospective survey was conducted to test the hypothesis that maximum anchorage can be achieved in the maxillary arch by controlling forces and moments using intraarch mechanics while retracting canines into first premolar extraction sites. The sample consisted of 24 patients (mean age 18 years, 9 months) who required the extraction of two maxillary first premolars, with or without extractions in the mandibular arch. Movements of the first molars, canines, and incisors were evaluated with 6 cephalometric variables and 10 study model variables. T tests were used to assess differences between pretreatment and postretraction tooth positions. Cephalometrically, the maxillary first molars (left and right sides combined) moved mesially ONLY 0.7 mm (SD 0.43; p < 0.008). All other cephalometric variables showed no significant differences between the two time points. From the study models, the molars moved mesially ONLY 0.5 mm on both the right and left sides (right side SD = 0.43 and left side SD = 0.38; p < 0.005), while the canines were retracted on average 5.8 mm on the right side and 5.6 mm on the left. The molars and canines showed significant mesiopalatal and distolingual rotations, respectively. Many of the study model and cephalometric variables were significantly correlated to one another. This study questions the need to use adjunctive appliances, which directs a distal force to the posterior teeth, if horizontal molar anchorage control is a treatment objective. By controlling forces and moments, using intraarch mechanics while retracting maxillary canines into first premolar extraction sites, minimal molar anchorage loss occurred. PMID- 9345158 TI - Lower arch perimeter preservation using the lingual arch. AB - The purpose of this investigation was to determine whether the placement of a mandibular lingual arch maintained arch perimeter in the transition from the mixed to the permanent dentition, and if so, whether it was effective at preventing mesial migration of first permanent molars, or whether this migration still occurred en masse, by increased lower incisor proclination. Thirty patients were randomly assigned to either a treatment group (N = 14, mean age = 11.5 years) or a control group (N = 16, mean age = 11.3 years). Study models, cephalograms, and tomograms of the patients, taken at the beginning and at the end of the study period, were examined. Statistically significant differences between groups were found for positional changes of mandibular first molars and incisors, and changes in arch dimensions. The results indicate that the lingual arch can help reduce arch perimeter loss, but at the expense of slight mandibular incisor proclination. PMID- 9345159 TI - Scanning devices for digitizing images. PMID- 9345160 TI - Litigation, legislation, and ethics. Patient noncooperation: contributory negligence or mitigation of damages? PMID- 9345161 TI - Measurement of airway patency. A manual for users of the Toronto systems and others interested in nasal patency measurement. PMID- 9345162 TI - Optimising the investigation of meningococcal disease. PMID- 9345164 TI - Peer review: reform or revolution? PMID- 9345163 TI - Climate change--thinking widely, working locally, acting personally. PMID- 9345165 TI - Diagnostics in developing countries. PMID- 9345166 TI - Determining prognosis after acute myocardial infarction in the thrombolytic era. PMID- 9345167 TI - Mortality associated with HIV-1 infection over five years in a rural Ugandan population: cohort study. AB - OBJECTIVE: To assess the impact of HIV-1 infection on mortality over five years in a rural Ugandan population. DESIGN: Longitudinal cohort study followed up annually by a house to house census and medical survey. SETTING: Rural population in south west Uganda. SUBJECTS: About 10,000 people from 15 villages who were enrolled in 1989-90 or later. MAIN OUTCOME MEASURES: Number of deaths from all causes, death rates, mortality fraction attributable to HIV-1 infection. RESULTS: Of 9777 people resident in the study area in 1989-90, 8833 (90%) had an unambiguous result on testing for HIV-1 antibody; throughout the period of follow up adult seroprevalence was about 8%. During 35,083 person years of follow up, 459 deaths occurred, 273 in seronegative subjects and 186 in seropositive subjects, corresponding to standardised death rates of 8.1 and 129.3 per 1000 person years. Standardised death rates for adults were 10.4 (95% confidence interval 9.0 to 11.8) and 114.0 (93.2 to 134.8) per 1000 person years respectively. The mortality fraction attributable to HIV-1 infection was 41% for adults and was in excess of 70% for men aged 25-44 and women aged 20-44 years. Median survival from time of enrollment was less than three years in subjects aged 55 years or more who were infected with HIV-1. Life expectancy from birth in the total population resident at any time was estimated to be 42.5 years (41.4 years in men; 43.5 years in women), which compares with 58.3 years (56.5 years in men; 60.5 years in women) in people known to be seronegative. CONCLUSIONS: These data confirm that in a rural African population HIV-1 infection is associated with high death rates and a substantial reduction in life expectancy. PMID- 9345168 TI - HIV antibody assay that gave false negative results: multicentre collaborative study. AB - OBJECTIVE: To identify false negative results arising from the use of a commercial kit to detect antibody to HIV-1 and HIV-2 between July 1995 and March 1996. DESIGN: The 56 laboratories in the United Kingdom that were using the assay were asked to retrieve and retest specimens with an alternative assay for HIV-1 and HIV-2. Details of false negative results were obtained and these serum samples further investigated. SUBJECTS: 24,181 patients tested with the assay who were reported as being negative for HIV antibody. An additional 497 patients were confirmed as HIV positive with the assay. RESULTS: Serum samples of 20,973 of the patients were retested, and four patients were found to have had false negative results with the kit; three further patients were found to have had false negative results in the course of other laboratory testing. The seven patients with false negative results with the kit were of diverse risk group and HIV-1 subtype. Four had evidence of recent HIV infection. CONCLUSION: The commercial kit had a sensitivity of 99.2% (497/501), or less if the additional three patients with false negative results were taken into account. PMID- 9345169 TI - Epidemiology and clinical management of meningococcal disease in west Gloucestershire: retrospective, population based study. AB - OBJECTIVE: To study changes in the epidemiology and management of meningococcal disease in one health district during a period of high local incidence of disease. DESIGN: Prospective case ascertainment and data collection over 14 years, with retrospective analysis of cases. SETTING: West Gloucestershire (population 320,000). SUBJECTS: Residents developing meningococcal disease between 1 January 1982 and 31 December 1995. RESULTS: 252 cases of invasive meningococcal disease were identified, of which 102 (40%) were officially notified and 191 (76%) were confirmed by culture from a deep site. The observed disease incidence of 5.6/100,000/year was about 2.7 times the national incidence (as measured by either statutory notifications or reference laboratory reports). The period 1983-90 was characterised by a prolonged localised outbreak due to serogroup B serotype 15 sulphonamide resistant (B15R) strains. General practitioners gave benzylpenicillin before hospital admission to 18% of patients who presented with meningococcal disease in the first half of the study period and to 40% who presented in the second half. The overall case fatality rate was 6.7% (17/252). Four deaths were directly or indirectly related to lumbar puncture. Of 120 patients whose lumbar puncture yielded meningococci, nine (8%) showed no abnormality on initial examination. CONCLUSIONS: Neither laboratory records nor formal notifications alone can give an accurate estimate of the incidence of meningococcal disease. Because of the dangers of lumbar puncture, the frequency of misleading negative initial findings, and the advent of new diagnostic techniques, the need for samples of cerebrospinal fluid should be critically questioned in each case of suspected meningococcal disease. PMID- 9345171 TI - Audit of process of antenatal screening for sickle cell disorders at a north London hospital. PMID- 9345170 TI - Audit of prenatal diagnosis for haemoglobin disorders in the United Kingdom: the first 20 years. AB - OBJECTIVES: To audit services for prenatal diagnosis for haemoglobin disorders in the United Kingdom. DESIGN: Comparison of the annual number of cases recorded in a United Kingdom register of prenatal diagnoses for haemoglobin disorders, with the annual number of pregnancies at risk of these disorders, by ethnic group and regional health authority. The number of pregnancies at risk was estimated using data on ethnic group from the 1991 census and data from the United Kingdom thalassaemia register, which records the number of babies born with thalassaemia. SETTING: The three national prenatal diagnosis centres for haemoglobin disorders. SUBJECTS: 2068 cases of prenatal diagnosis for haemoglobin disorders in the United Kingdom from 1974 to 1994. MAIN OUTCOME MEASURES: Utilisation of prenatal diagnosis by risk, ethnic group, and regional health authority. Proportion of referrals in the first trimester and before the birth of any affected child. RESULTS: National utilisation of prenatal diagnosis for haemoglobin disorders was around 20%. During the past 10 years it has remained steady at about 50% for thalassaemias and risen from 7% to 13% for sickle cell disorders. Utilisation for sickle cell disorders varies regionally from 2% to 20%. Utilisation for thalassaemias varies by ethnic group. It is almost 90% for Cypriots and ranges regionally for British Pakistanis from 0% to over 60%. About 60% of first prenatal diagnoses are done for couples without an affected child. Less than 50% of first referrals are in the first trimester. CONCLUSIONS: National utilisation of prenatal diagnosis for haemoglobin disorders is far lower than expected, and there are wide regional variations. A high proportion of referrals are still in the second trimester and after the birth of an affected child. The findings point to serious shortcomings in present antenatal screening practice and in local screening policies and to inadequate counselling resources, especially for British Pakistanis. PMID- 9345172 TI - Intranasal chlorhexidine resulting in anaphylactic circulatory arrest. PMID- 9345173 TI - Effects on birth weight and perinatal mortality of maternal dietary supplements in rural Gambia: 5 year randomised controlled trial . AB - OBJECTIVE: To test the efficacy in terms of birth weight and infant survival of a diet supplement programme in pregnant African women through a primary healthcare system. DESIGN: 5 year controlled trial of all pregnant women in 28 villages randomised to daily supplementation with high energy groundnut biscuits (4.3 MJ/day) for about 20 weeks before delivery (intervention) or after delivery (control). SETTING: Rural Gambia. SUBJECTS: Chronically undernourished women (twin bearers excluded), yielding 2047 singleton live births and 35 stillbirths. MAIN OUTCOME MEASURES: Birth weight; prevalence of low birth weight (< 2500 g); head circumference; birth length; gestational age; prevalence of stillbirths; neonatal and postneonatal mortality. RESULTS: Supplementation increased weight gain in pregnancy and significantly increased birth weight, particularly during the nutritionally debilitating hungry season (June to October). Weight gain increased by 201 g (P < 0.001) in the hungry season, by 94 g (P < 0.01) in the harvest season (November to May), and by 136 g (P < 0.001) over the whole year. The odds ratio for low birthweight babies in supplemented women was 0.61 (95% confidence interval 0.47 to 0.79, P < 0.001). Head circumference was significantly increased (P < 0.01), but by only 3.1 mm. Birth length and duration of gestation were not affected. Supplementation significantly reduced perinatal mortality: the odds ratio was 0.47 (0.23 to 0.99, P < 0.05) for stillbirths and 0.54 (0.35 to 0.85, P < 0.01) for all deaths in first week of life. Mortality after 7 days was unaffected. CONCLUSION: Prenatal dietary supplementation reduced retardation in intrauterine growth when effectively targeted at genuinely at-risk mothers. This was associated with a substantial reduction in the prevalence of stillbirths and in early neonatal mortality. The intervention can be successfully delivered through a primary healthcare system. PMID- 9345174 TI - Evaluation of computer support for prescribing (CAPSULE) using simulated cases. AB - OBJECTIVE: To evaluate the potential effect of computer support on general practitioners' prescribing, and to compare the effectiveness of three different support levels. DESIGN: Crossover experiment with balanced block design. SUBJECTS: Random sample of 50 general practitioners (42 agreed to participate) from 165 in a geographically defined area of Oxfordshire. INTERVENTIONS: Doctors prescribed for 36 simulated cases constructed from real consultations. Levels of computer support were control (alphabetical list of drugs), limited support (list of preferred drugs), and full support (the same list with explanations available for suggestions). MAIN OUTCOME MEASURES: Percentage of cases where doctors ignored a cheaper, equally effective drug; prescribing score (a measure of how closely prescriptions matched expert recommendations); interview to elicit doctors' views of support system. RESULTS: Computer support significantly improved the quality of prescribing. Doctors ignored a cheaper, equally effective drug in a median 50% (range 25%-75%) of control cases, compared with 36% (8%-67%) with limited support and 35% (0-67%) with full support (P < 0.001). The median prescribing score rose from 6.0 units (4.2-7.0) with control support to 6.8 (5.8 to 7.7) and 6.7 (5.6 to 7.8) with limited and full support (P < 0.001). Of 41 doctors, 36 (88%) found the system easy to use and 24 (59%) said they would be likely to use it in practice. CONCLUSIONS: Computer support improved compliance with prescribing guidelines, reducing the occasions when doctors ignored a cheaper, equally effective drug. The system was easy to operate, and most participating doctors would be likely to use it in practice. PMID- 9345175 TI - Social phobia: epidemiology, recognition, and treatment. PMID- 9345176 TI - ABC of palliative care. Principles of palliative care and pain control. PMID- 9345177 TI - Global climate change: the potential effects on health. PMID- 9345178 TI - Hyperglycaemia after acute stroke. Other models find that hyperglycaemia is not independent predictor. PMID- 9345179 TI - Hyperglycaemia after acute stroke. May occur as result of neuroendocrine response. PMID- 9345180 TI - Hyperglycaemia after acute stroke. Participants required for trial of treatment with glucose and insulin. PMID- 9345181 TI - More money is needed to care for patients with cancer. PMID- 9345182 TI - Cancer self help groups are underused. PMID- 9345183 TI - Case-control study of sudden infant death syndrome in Scotland. Income level or bed sharing would confound any effect of previous use of mattress. PMID- 9345184 TI - Case-control study of sudden infant death syndrome in Scotland. Multiple statistical comparisons used are confusing. PMID- 9345185 TI - Case-control study of sudden infant death syndrome in Scotland. Risk of bed sharing was not sufficiently examined. PMID- 9345186 TI - Giving thyroid hormones to clinically hypothyroid but biochemically euthyroid patients. Supporting authors' views would be unwise. PMID- 9345187 TI - Giving thyroid hormones to clinically hypothyroid but biochemically euthyroid patients. Distinguishing hypothyroid symptoms from common non-specific complaints is difficult. PMID- 9345188 TI - Giving thyroid hormones to clinically hypothyroid but biochemically euthyroid patients. Long-term treatment is being used. PMID- 9345189 TI - Diagnosing pulmonary embolism. PMID- 9345190 TI - Pneumococcal vaccine campaign based in general practice. Further prospective randomised controlled trial is necessary. PMID- 9345191 TI - Pneumococcal vaccine campaign based in general practice. Expert advice is unclear. PMID- 9345192 TI - Pneumococcal vaccine campaign based in general practice. Datasheet for vaccine contradicts authors' recommendations. PMID- 9345193 TI - Pneumococcal vaccine campaign based in general practice. Small audit showed that only 14% of patients were offered pneumococcal vaccine. PMID- 9345194 TI - Managing eye conditions in general practice. PMID- 9345195 TI - Medical managers. Doctors need training in management skills. PMID- 9345196 TI - Medical managers. BMA is also working to develop role of clinicians in management. PMID- 9345197 TI - Some orthopaedic findings in ninety-eight cases of hemophilia. 1936. PMID- 9345198 TI - Pathogenesis, early diagnosis, and prophylaxis for chronic hemophilic synovitis. AB - The most common clinical manifestation of hemophilia is intraarticular bleeding (hemarthrosis). Although any joint can be involved, the articulations most frequently affected are the knees, elbows, and ankles. Hemorrhages may occur spontaneously, or as a result of trauma. Because hemophilia is a genetically determined coagulation disorder, the bleeding incidences may start early in a child's life. After several hemarthroses, hemophilic synovitis will develop and, because of the profusion and fragility of vessels within the tissue, will lead to a vicious cycle of bleeding-synovitis-bleeding. If the synovitis is not controlled, cartilage damage will occur. hemophilic arthropathy then will become apparent clinically and radiographically, causing functional impairment of the affected joint. Prophylaxis and early diagnosis of acute hemarthrosis are essential to prevent the development of hemophilic synovitis. Hemarthroses must be treated aggressively if hemophilic synovitis is to be prevented. Continuous prophylaxis from age 2 to 18 years has been claimed to reduce the incidence of chronic hemophilic synovitis and joint damage, which remain the main causes of disability in the life of the patient who has hemophilia. PMID- 9345199 TI - Conservative treatment of hemarthrosis for prevention of hemophilic synovitis. AB - Acute hemarthroses are probably the most frequent type of bleeding in the patient with hemophilia. Delayed and/or inadequate treatment can trigger a series of pathologic changes within the joint leading to a painful and disabling arthropathy. Despite the advent of prophylactic treatment with factor concentrates, the majority of patients in the world have no access to even on demand factor replacement. Care for all patients involves a team approach led by the hematologist but including input from orthopaedic surgeons and physiotherapists. Optimal treatment involves a combination of factor replacement, rest, ice, and supervised rehabilitation. In certain cases, joint aspiration may be considered. In developing countries, where factor concentrates are in short supply, such bleeding episodes usually are treated by physical means alone or with the addition of cryoprecipitate or fresh frozen plasma. After successful resolution of such episodes by whatever means, the events leading to the bleeding episode and its subsequent management should be considered within the setting of the treating unit. Such debriefings should aim to provide counsel regarding any appropriate lifestyle modifications and, where necessary, treatment should be arranged to minimize the risk of additional episodes. PMID- 9345200 TI - Prophylactic transfusion for hypertrophic synovitis in children with hemophilia. AB - A 7- to 9-month protocol of prophylactic transfusion was used to treat 33 joints in 19 children with severe hemophilia (< 1 U/dL Factors VIII or IX) and hypertrophic synovitis. The overall rate of hemarthrosis was reduced, but only 36% (12 of 33 joints) achieved a good result (defined as 0-0.5 bleeding episodes per month and decreased synovial hypertrophy 1 year after completing treatment). Age and severity of arthropathy at initiation of treatment did not affect the result. The degree of synovial hypertrophy and involvement of the knee joint showed an adverse trend, but these factors did not achieve statistical significance. The number of episodes of breakthrough bleeding during the first 6 weeks of therapy was significantly associated with a poor result. Based on the results of this study, a trial of transfusion therapy is recommended for recurrent hemarthroses and synovitis in patients with hemophilia, but the duration of thrice weekly treatment has been increased and the duration of prophylaxis has been reduced in selected cases. PMID- 9345201 TI - Intraarticular dexamethasone in advanced chronic synovitis in hemophilia. AB - From 1988 to 1966, 34 patients with advanced chronic hemophilic synovitis (25 Grade III and nine Grade IV) were treated with intraarticular injections of long acting dexamethasone (sodium phosphate of dexamethasone plus acetate of dexamethasone) in cycles of three injections with 3-week intervals between each injection with 6-month rest intervals between cycles for as many as three cycles, depending on the evolution of each case. All patients had chronic severe synovitis, axial deformity, muscular atrophy, and diminution of range of movement. There were 31 knees, two ankles, and two shoulders. Subjective and objective evaluations were done grouping the results in good, fair, and poor according to grade of patient satisfaction, presence of synovitis and pain, range of movement, and limitation of activities of daily living. In the subjective results there were 19 good results, 12 fair results, and four poor results, and in the objective evaluation there were 22 good results, nine fair results, and four poor results at an average followup of 1.5 years. The use of intraarticular dexamethasone as an alternative in the short to medium term for treatment of advanced chronic hemophilic synovitis with pain and limitation of function before doing an invasive surgical treatment is proposed. PMID- 9345202 TI - Chemical synoviorthesis for hemophilic synovitis. AB - For many years, Rifampicin has been used empirically for the treatment of hemophilic chronic synovitis with encouraging results. A study was performed in which Rifampicin was shown to reduce the inflammation of joints affected by hemophilic synovitis. A clinical study was performed on 48 hemophilic patients (48 joints). Seventeen elbows, eight knees, and 23 ankles were treated. The mean age of the patients was 6 years (range, 4-23 years) and the mean followup was 29 months (range, 24-53 months). Overall, 40 excellent results and eight good results were obtained. The average number of weekly injections of Rifampicin was 3.06 (range, 1-10 injections). Eight patients experienced pain on the first injection, which subsided gradually with the subsequent procedures. Synoviorthesis with Rifampicin seems to be a good method for the treatment of hemophilic synovitis, especially in small joints (elbows and ankles) and in younger children. PMID- 9345203 TI - Radioactive synoviorthesis in patients with hemophilia with factor inhibitor. AB - In nine patients with hemophilia and factor inhibitor (six with hemophilia A; three with hemophilia B), 19 joints were treated with radioactive synoviorthesis using Au-198. Ages ranged from 3 to 40 years. Synoviorthesis was performed when the antibody titer was low (< 10 Bethesda units), thus making hemostasis possible by factor administration for 2 to 4 days. On five occasions, radioactive synoviorthesis was performed simultaneously with tolerance induction according to the Malmo protocol. A bleeding free interval of more than 6 months was obtained in 11 joints, six of which remained bleeding free for more than a year. At long term followup (range, 18-182 months) five joints were rated good, one joint was fair, and 11 joints were poor. Although the results are inferior to those for patients with hemophilia without inhibitor, radioactive synoviorthesis should be considered because of its ease of performance and the definite decrease in joint bleeding frequency that it brings about. This is of particular interest in patients with hemophilia caused by factor inhibitor who otherwise are difficult to treat. PMID- 9345204 TI - Physiotherapy for prevention and treatment of chronic hemophilic synovitis. AB - Hemophilia is a hereditary lifelong bleeding disorder affecting males. The nature of the condition predisposes the person to bleed intraarticularly and intermuscularly. Without intervention with replacement factor therapy and physiotherapy, the consequences can lead to chronic synovitis and severe joint hemarthropathy. Physiotherapeutic interventions are available that may help to prevent and treat the sequelae of recurrent hemarthrosis. No particular method of treatment has been shown to eradicate hemorrhages; however, there are protocols that can help. The use of physiotherapy techniques including electrotherapy, joint care, and exercise are extremely important, especially in developing countries where blood products are scarce. PMID- 9345205 TI - Hemophilic synovitis of the knee and the elbow. AB - A prospective study from 1974 to 1996 was done to determine optimal treatment for chronic hemophilic synovitis of the knee and synovitis of the elbow. Sixty-five patients with synovitis affecting 65 knee joints and 40 patients who had synovitis of the elbow (44 elbows), despite a 3-month trial of prophylactic substitution therapy, were treated by synovectomy. Radiation synovectomies (Au 198 synoviorthesis) were done on 38 knees, open surgical synovectomy on 18, and nine had an arthroscopic procedure. Radioactive gold synoviorthesis was performed on 29 elbows, and 15 had a resection of the radial head and partial open synovectomy. Synovectomy (by any method) significantly reduced bleeding episodes, but did not halt the radiographic deterioration of the joints. It is thought that radiation synovectomy is the best choice for patients with persistent synovitis of the knee and synovitis of the elbow unresponsive to a 3-month trial of prophylactic factor replacement. If two to three consecutive synoviortheses with 3 to 6 months intervals had been ineffective, or when the radiographic score is more than two points, an open synovectomy is indicated. PMID- 9345206 TI - Orthotic management of the knee in patients with hemophilia. AB - The knee joint is the most frequently affected articulation in patients suffering from hemophilia. Before the availability of active coagulation factors, orthotic use was aimed at corrective functions. Orthotics in the modern era play a protective and preventative role, and they are used in the treatment of hemarthroses and in joints affected by chronic synovitis. Because levels of availability of clotting factors and orthotic material vary extensively world wide, an overview is provided whereby each group of therapists can adapt themselves to the principles. PMID- 9345207 TI - Arthroscopy for chronic hemophilic synovitis of the knee. AB - Between 1988 and 1995, 32 knee joints (29 patients) with hemophilic arthropathy underwent arthroscopy. The spectrum of procedures ranged from resection of fibrous plicae to synovectomy. Technical difficulties appeared in cases of scarred fixed patella, pronounced posterior tibia subluxation, and severe fibrous ankylosis. All operations done between 1988 and 1991 (23 operations; 21 patients) were reviewed retrospectively. The mean age of the patients in this series was 30 years and the mean followup was 5 years. On subjective evaluation, 13 operations achieved a definite improvement, five showed slight improvement, and two had no improvement (three operations were excluded). Arthroscopic surgery, as a relatively low risk technique, combined with early functional rehabilitation, can be used to achieve satisfactory results in patients with hemophilic arthropathy. PMID- 9345208 TI - Hamstring release for fixed knee flexion contracture in hemophilia. AB - Sixteen hamstring tenotomies and posterior capsulotomies were assessed retrospectively in 10 patients with hemophilia. The average age of the patients was 17 years (range, 16-24 years). The main indication for surgery was a fixed knee flexion contracture of 30 degrees to 45 degrees, associated with repeated hemarthroses, and failure of conservative treatment after 6 months. A posterior transverse capsulotomy and a Z shaped hamstring tenotomy were performed. The postoperative treatment consisted of repeated stretching exercises for a 6-month period. An average decrease of 25 degrees in the amount of fixed knee flexion contracture was obtained (range, 10 degrees-40 degrees). Followup for an average of 9.5 years showed 11 good, four fair, and one poor result regarding their joint scores. It is concluded that hamstring release is an effective surgical procedure for fixed knee flexion contracture in hemophilia. It seems to reduce the incidence of hemarthrosis relating to such a deformity. PMID- 9345209 TI - Orthotics and rehabilitation for chronic hemophilic synovitis of the ankle. An overview. AB - The ankle is the second most affected joint in hemophilia. Recurrent bleeding leads to chronic synovitis. Prevention of chronic synovitis should start with prophylactic replacement therapy or on demand treatment of recurrent hemarthrosis. Attention must be given to the function of the ankle joint. When instability is present, an extensive range of exercises is important. An orthotic or shoe adaption may be useful during the rehabilitation process. Viscoheels worn in shoes seem to reduce the bleeding frequency and pain. PMID- 9345210 TI - Gait analysis of the hemophilic ankle with silicone heel cushion. AB - Shock absorption becomes very important in damaged joints with destroyed cartilage and progressive muscular imbalance as occurs in hemarthropathy. The effects of silicone heel cushioning on the ankle motion of hemophilic patients in different stages of hemarthropathy of the ankle joints was measured using an ultrasound motion analysis system. It is concluded that silicone heel cushioning has no influence on ankles in the late stage of hemarthropathy. Silicone heel cushioning will lead to uncontrolled changes of the ankle joint in the early hemarthropathic ankle, involving the tibiotalar and the subtalar joints. The angular velocity of the ankle is increased producing higher acceleration at the ankle joint. The higher angle acceleration is related to higher joint loading uncontrolled by the muscles. The resulting uncoordinated motion can cause ligamentous overloading, strains, and a higher probability of joint bleeding. Therefore, silicone heel cushioning or other shock absorbing devices that return the energy immediately to the foot are not useful for prevention and treatment of chronic hemophilic synovitis and may cause additional deterioration of the joint. PMID- 9345211 TI - Orthopaedic surgery in hemophilic patients with human immunodeficiency virus. AB - Patients registered at the author's hemophilia center between 1982 and 1994 were studied to establish whether major orthopaedic surgical procedures accelerate the fall of CD4 lymphocyte counts of patients with hemophilia who are infected with the human immunodeficiency virus, and whether patients who had surgery had different rates of development of acquired immune deficiency syndrome or death when compared with patients who did not have surgery. The patients were divided into four groups: Group 1, 22 patients who were human immunodeficiency virus positive undergoing orthopaedic surgery; Group 2, 89 patients who were human immunodeficiency virus positive not undergoing orthopaedic surgery; Group 3, 18 patients who were human immunodeficiency virus negative undergoing orthopaedic surgery; and Group 4, 135 patients who were human immunodeficiency virus negative not undergoing orthopaedic surgery. There was no significant difference between the rates of decline of CD4 lymphocyte counts for patients who were human immunodeficiency virus positive who underwent surgery when compared with human immunodeficiency virus positive patients who did not undergo surgery, nor was there any significant difference between the two human immunodeficiency virus negative groups. There were no significant differences in the rate of development of acquired immune deficiency syndrome or mortality rates between patients who had surgery and those who did not. PMID- 9345212 TI - Therapeutic options in the management of hemophilic synovitis. AB - There is a complex relationship between recurrent bleeding, synovitis, and the development of arthritis in the patient with hemophilia. There are many options available for the treatment of recurrent bleeding and hemophilic synovitis, indicating that none works very well. Conservative treatment, including replacement of the missing clotting factor for 3 to 6 months, intermittent steroids, immobilization, and physical therapy should be tried before synovectomy is indicated. Synovectomy can be achieved through an open procedure, arthroscopically, or by injection of a radioactive material into the joint. Radioactive synovectomy is indicated in patients with inhibitors to the clotting factor, patients with advanced human immunodeficiency virus and advanced hepatitis, and in those patients with multiple joint involvement. Arthroscopic synovectomy is the procedure the authors recommend for the knee and ankle joints, although open synovectomy offers an excellent alternative. The greatest risk to these procedures is a decreased range of motion, and this is most problematic in the young child who cannot cooperate with a program of physical therapy. PMID- 9345213 TI - Surgical treatment of postoperative deltoid origin disruption. AB - Although it is well recognized that deltoid disruption after shoulder surgery is associated with poor function, little information is available regarding results of surgical treatment for this problem. Twenty-four patients underwent direct repair or rotational deltoidplasty reconstruction of a detached muscle origin after shoulder surgery. The original surgical procedure was rotator cuff repair in 12, acromioplasty in four, and lateral acromionectomy with or without rotator cuff repair in eight. The average duration of symptoms before deltoid reconstruction was 17 months. The mean followup was 39 months (range, 13-84 months). Twelve patients reported moderate to severe pain, whereas 12 had minimal pain. Two patients required a shoulder fusion for intractable pain. Overall, one (4%) excellent, seven (29%) good, and 16 (67%) unsatisfactory results were observed. A poor outcome was associated with a prior lateral acromionectomy, involvement of the middle deltoid, a massive rotator cuff tear with weakness in external rotation, and a residual postoperative defect larger than 2 cm. In select cases, repair or deltoidplasty can improve function and pain. PMID- 9345215 TI - Russell's sign. Subtle hand changes in patients with bulimia nervosa. AB - Bulimia nervosa is a common eating disorder, affecting between 1% to 10% of adolescent girls and college aged women. Because excessive weight loss and amenorrhea are not significant features, as they are in anorexia, bulimia is much harder to diagnose. Orthopaedic surgeons have a unique opportunity to detect one of the few physical signs of the disease, which is skin lesions, consisting of abrasions, small lacerations, and callosities on the dorsum of the hand overlying the metacarpophalangeal and interphalangeal joints. These nondescript dorsal lesions are caused by repeated contact of the incisors to the skin of the hand that occur during self induced vomiting. This finding, known as Russell's sign, may be seen by orthopaedic surgeons during examinations for other reasons. Because eating disorders are recognized as a component of the female athlete triad of osteoporosis, amenorrhea, and eating disorders and because orthopaedic surgeons routinely care for female athletes susceptible to these disorders, recognizing this sign and its implications may have profound influence on the patient's musculoskeletal system and general health. PMID- 9345214 TI - Herbert screw fixation for scaphoid nonunions. An analysis of factors influencing outcome. AB - A retrospective review of 160 cases of scaphoid nonunion treated by internal fixation using a Herbert screw with bone grafting was conducted at an average followup of 24 months. Definite radiographic union was achieved in 90% of cases. Based on Cooney's clinical scoring system, 80 cases had an excellent result, 37 had a good result, 33 had a fair result, and 10 had a poor result. Failure of union was related to the existence of avascular changes of the proximal fragment, instability of the fracture fragment, the prolonged delay in surgery, and the location of the fracture site. In the united scaphoids, the lengthy period of postoperative immobilization, the existence of osteoarthritis, and the prolonged delay in surgery were significant factors in the patient's functional outcome. Overall, the results do not support the view that a residual flexion deformity of the scaphoid is less likely to yield a satisfactory outcome, although it seems worthwhile to correct excessive angulation at the time of repair to promote an anatomic union, thereby preventing early arthritis. A bone graft with internal fixation using a Herbert screw and a shorter period of immobilization may give a satisfactory functional result when the nonunion is treated before the onset of arthritic changes in the wrist. PMID- 9345216 TI - Rotation osteotomies for osteonecrosis of the femoral head. AB - Twenty consecutive rotation osteotomies for idiopathic necrosis of the femoral head with an average followup of 6.5 years were reviewed. The original technique used a nail plate for rotation and fixation of the fragments and proved to be reliable for precision of rotation, osteotomy fusion, and absence of mechanical or vascular complications. There were 16 anterior extension Sugioka osteotomies with 52 degrees average rotation, and four posterior flexion Kempf osteotomies with 77 degrees average rotation. The status of 18 surgically treated hips after 5 years was seven failures, two fair results, and nine satisfactory results. Rotation osteotomies are recommended only when the necrotic zone of the femoral head can be removed from the major weightbearing zone of the acetabulum, when the hip is in extension. In Sugioka's anterior rotation, the necrotic zone, although unloaded in extension, usually remains in contact with the acetabular major bearing zone (40 degrees around apex) in hip flexion. Thus, it is recommended only for Ficat Stage 2 nonflattened heads. In Kempf's posterior rotation, the necrotic zone is unloaded in hip extension and flexion, so Stage 3 is not a contraindication for this osteotomy. In addition, osteotomies are not recommended when the necrosis extends deeper than the proximal third of the femoral head. In these large necroses, there may be an overloading of the healthy part of the rotated head, resulting in its secondary mechanical deterioration. If these conditions are fulfilled, rotation osteotomy of the proximal femur may, in patients younger than 45 years of age, delay for a decade the degradation of hips with idiopathic osteonecrosis. PMID- 9345217 TI - Quantifying osteoarthrotic hip incongruence. An approach to optimizing osteotomies. AB - Osteoarthrosis of the hip may be treated by osteotomy, but surgeons report variable results, and there is no consensus regarding which method to use in choosing the type of osteotomy. The authors defined three biomechanical measures of hip incongruence (characteristic point locus, joint space, and contact region) and developed a two-dimensional frontal plane model to compute joint incongruence over the joint range of motion during normal activities of daily living. The preoperative measures were calculated for 38 patients who had undergone osteotomy at least 5 years earlier. The authors calculated the measures throughout a functional range of motion after 13 stimulated varus or valgus osteotomies. A logistic regression analysis determined which, if any, of the three measures, in conjunction with other clinical variables, correctly predicted outcome. The average values for the characteristic point locus, joint space, and contact region measures ranged from 0.260 cm to 2.127 cm, 0.963 cm2 to 9.327 cm2, and 0.063 cm to 4.230 cm, respectively. Unimodal behavior between two of the three measures (joint space and contact region) and osteotomy angle were observed, suggesting these two would be the most useful in predicting an optimal osteotomy. The most significant independent variable predicting clinical outcome was the joint space measure. This supports the potential of an optimization approach for determining the best angle for a hip osteotomy. PMID- 9345219 TI - Three-dimensional kinematics of the human knee with intracortical pin fixation. AB - Knee motion was measured with an instrumented spatial linkage (accuracy, linear +/-500 microns; angular, +/-0.5 degree) fixed with intracortical Kirschner wires in five healthy male volunteers (five knees, judged clinically to be normal). This technique allows an accurate description of the relative angular and linear movements between tibia and femur without the effect of skin movement relative to the bone and without the effect of changing muscle volume. Motion of the tibia relative to the femur was described in terms of three clinically meaningful rotations and three translations between full extension and 60 degrees flexion: (1) abduction and adduction: 3.4 degrees +/- 1.2 degrees; (2) internal and external rotation: 10.6 degrees +/- 2.8 degrees, representing screw home motion; (3) anterior and posterior: 5.2 +/- 1.7 mm, representing roll back phenomenon; (4) proximal and distal: 1.2 +/- 2.7 mm; and (5) medial and lateral: 1.1 +/- 2.6 mm. PMID- 9345218 TI - A fully implantable motorized intramedullary nail for limb lengthening and bone transport. AB - This article describes an intramedullary nail that contains a fully implantable motorized programmable sliding mechanism for limb lengthening and bone transport that reduces the risk of infection, discomfort, and scarring usually associated with the external fixators used for the same purpose. Twelve patients were treated surgically with the new system. Eleven patients had unilateral femur shortening between 3 and 7.5 cm, and one patient had a 12-cm defect after tumor resection. In all patients with femur shortening the leg length discrepancy was corrected completely. In the case of bone defect the segment transport worked well without any problems. There was no infection and no axial deformity. Immediately after chemotherapy, delayed bone formation was seen. In two early cases of limb lengthening a technical problem led to replacement of the motor. PMID- 9345220 TI - Tarsal tunnel syndrome. Outcome of surgery in longstanding cases. AB - Cases of longstanding (median, 60 months) tarsal tunnel syndrome were decompressed surgically in 14 female and four male patients. Patients reported intermittent dysesthesia, paresthesia, or anesthesia at the medial plantar aspect of the foot. Symptoms were aggravated by physical activities. Previous trauma was noted in four patients. Tinel's sign was positive in 16 patients. Magnetic resonance imaging was performed in 10 patients but was conclusive in only two. At surgery, the posterior tibial nerve or one of its branches was found to be entrapped in 15 patients. Entrapments were observed isolated or in combination within the fascial septa (n = 5), varicose veins (n = 6), scar tissues (n = 4), tenosynovitis and edema (n = 1), or within the abductor hallucis muscle (n = 1). Two neuromas were excised. In three patients no obvious entrapments were found. Clinical followup was performed a median 18 months after surgery. Relief of symptoms was reported as long as 1 year after surgery. All symptoms were relieved in 11 (61%) patients. Three (17%) patients with previous trauma had relatively severe pain after surgery and were considered to have failed results. Surgical decompression was beneficial in most patients with longstanding tarsal tunnel syndrome. PMID- 9345221 TI - Short-term external support promotes healing in semirigidly fixed fractures. AB - The effect of different postoperative treatments on the healing of rabbit femoral shaft fractures fixed semirigidly by intramedullary nailing was investigated clinically, radiologically, and mechanically. A unilateral transverse midfemoral fracture was performed on all rabbits and was treated with an intramedullary nailing without previous reaming. When the operated limb was immobilized postoperatively by a splint with the knee in extension for 1 week (Group 2), nine of 10 fractures healed. When immobilized in extension for 7 weeks (Group 3), eight of 10 healed. When immobilized in flexion for 7 weeks (Group 4), five of 10 fractures were clinically healed within 7 weeks. When the limb was not immobilized by external support (Group 1), only one of 10 healed. The radiologic healing correlated with the clinical healing in each group. There were no statistically significant differences among the groups in the quantity of callus or in the biomechanical properties of the healed fractures. The results indicate that even a short term additional external support of the limb with the knee in extension was advantageous to the healing of femoral shaft fractures fixed semirigidly by intramedullary nailing. PMID- 9345222 TI - Total knee arthroplasty infections associated with dental procedures. AB - Total knee arthroplasties are at risk for hematogenous seeding secondary to procedures that create a transient bacteremia. To define the risk of infection associated with dental surgery, a retrospective review of the records of 3490 patients treated with total knee arthroplasty by the authors between 1982 and 1993 was performed. Sixty-two total knee arthroplasties with late infections (greater than 6 months after their procedure) were identified, and of these, seven infections were associated strongly with a dental procedure temporally and bacteriologically. These seven cases represented 11% of the identified infections or 0.2% of the total knee arthroplasty procedures performed during this period. In addition, among 12 patients referred for infected total knee arthroplasties from outside institutions, two infections were associated with a dental procedure. Five of the nine (56%) patients had systemic risk factors that predisposed them to infection, including diabetes and rheumatoid arthritis. All dental procedures were extensive in nature (average, 115 minutes; range, 75-205 minutes). Eight of the patients received no antibiotic prophylaxis. One patient had only one preoperative dose. Infections associated with dental procedures may be more common than previously suspected. Eight of these patients had no prophylactic antibiotics, and one had inadequate coverage. The authors think that patients with a total knee arthroplasty who have systemic disease that compromises host defense mechanisms against infections and who undergo extensive dental procedures should receive prophylactic antibiotics. A first generation cephalosporin, given 1 hour preoperatively and 8 hours postoperatively would provide the best prophylaxis against the organisms identified in this study. PMID- 9345223 TI - Ewing's sarcoma of the foot. AB - The results of treatment were reviewed in 16 patients (10 male and six female) who had Ewing's sarcoma of the foot from 1954 through 1992. Mean age was 17 years (range, 10-42 years). The tumor involved the metatarsals (six patients), phalanges (four), calcaneus (three), navicular (one), talus (one), and calcaneus and phalanx (one). Seven patients had metastatic disease at the time of diagnosis, and only one of these patients survived. None of the patients with pulmonary metastasis at presentation survived. Nine patients had localized disease at the time of diagnosis, and eight survived. In the overall series, nine of the 16 patients were alive at followup (eight survived at least 5 years). Diagnosis was established at an average of 14 months from the onset of symptoms: 7 months in forefoot tumors and 22 months in hindfoot tumors. None of the six patients who had a resection had local failure. Seven of the 10 patients with forefoot lesions survived, and two of the six patients with hindfoot lesions survived. Treatment of Ewing's sarcoma of the foot by local control with radiation or operation and systemic control with chemotherapy is recommended. Survival appears to be better in patients who present with localized disease and forefoot lesions. Survival is worse in patients who present with metastatic disease. Surgical treatment appears to have an important role in local control and survival. PMID- 9345224 TI - Bone bonding ability of bioactive bone cements. AB - The bone bonding ability of three types of bioactive bone cement A, B, and C consisting of glass or glass ceramic powder and bisphenol-alpha-glycidyl methacrylate resin was evaluated. Type A contained MgO-CaO-SiO2-P2O5-CaF2 glass powder; Type B, MgO-CaO-SiO2-P2O5-CaF2 glass ceramic powder; and Type C, MgO free CaO-SiO2-P2O5-CaF2 glass powder. Rectangular plates (2 x 10 x 15 mm) of Types A, B, C, and polymethylmethacrylate cements were implanted into the tibial metaphyses of male rabbits and the failure load measured by mechanical failure testing (detaching test) 10 and 25 weeks after implantation. The failure loads of Types A, B, C, and polymethylmethacrylate cements were respectively, 29.52, 41.48, 28.22, and 0.29 N at 10 weeks and 33.42, 41.27, 33.64, and 0.20 N at 25 weeks. Examination of the bone cement interface revealed that all the bioactive bone cements achieved direct bone contact with the bone. These results showed that all three types of bioactive bone cement have the ability to bond to bone, and the cement containing glass ceramic powder revealed higher bonding strength than did those containing glass powder. PMID- 9345226 TI - Graft healing after anterior cruciate ligament reconstruction in rabbits. AB - Anterior cruciate ligament reconstruction with patellar tendon was performed in 50 rabbits (two groups of 25 animals) by the outside-in (Group I) and the inside out (Group II) techniques. Five animals from each group were sacrificed at different times (2 weeks, 1, 3, 6, and 9 months). Histologic analysis showed that the intraarticular part of the graft was morphologically similar to a normal ligament in both groups at 9 months. In Group 1, a newly formed bone-graft junction along the tunnel walls was observed inside the femoral tunnel. At 6 months, this junction resembled a direct type junction. The old bone-tendon junction showed an early disappearance of the fibrocartilage and was differentiated as a direct junction only at 9 months. In Group 2, at the site of the old bone-tendon junction a fibrocartilaginous layer was present during the whole process of remodeling, and at 6 months this area resembled a direct junction. These observations would suggest that when the junction is placed inside the tunnel (outside-in technique) the process of remodeling is more dramatic and slower than when it is placed at the intraarticular exit of the tunnel (inside-out technique), probably because of the formation of a new bone graft junction along the tunnel walls that partially unload the old junction. PMID- 9345227 TI - In vivo study of stainless steel and Ti-13Nb-13Zr bone plates in a sheep model. AB - A sheep study was performed to compare the in vivo performance of bone plates of 316L stainless steel and a new titanium alloy, titanium + 13% niobium + 13% zirconium (Ti-13Nb-13Zr), which had been subjected to a diffusion hardening treatment to produce a blue, wear resistant surface. Bone plates and screws of stainless steel and diffusion hardened Ti-13Nb-13Zr were implanted in adult sheep, in one group (with unosteotomized femurs) for 16 weeks, and in the other (with osteotomized femurs) for 8 weeks. At harvest, the diffusion hardened Ti 13Nb-13Zr devices had superior fixation strength, with greater screw torque out strength and fewer loose screws. In the osteotomized animals, the femurs with diffusion hardened Ti-13Nb-13Zr plates had higher torsional strength after removal of the implants; however, the difference was not statistically significant. In the unosteotomized animals, the torsional strength of the femurs was identical for both materials. There was a slightly reduced incidence of infection (bacterial adhesion) for the sheep with diffusion hardened Ti-13Nb-13Zr implants. In a parallel in vitro study, the magnetic resonance imaging compatibility of Ti-13Nb-13Zr was significantly superior to that of stainless steel. This indicates that diffusion hardened Ti-13Nb-13Zr may be an attractive alternative material for osteosynthesis. PMID- 9345225 TI - Slowly progressive osteoarthritis after tibial valgus osteotomy in young beagle dogs. AB - A slowly progressive osteoarthritis model in the skeletally immature canine knee joint is described. Forty-four young female beagle dogs were chosen as experimental animals. In 15 dogs, a 30 degrees valgus angulation of the right tibia was created by operation. Fourteen dogs underwent sham operation. Fifteen dogs served as control subjects. Alterations in the knee joints were evaluated macroscopically and histologically 7 and 18 months after operation. Seven months after surgery, two of seven beagles that had valgus osteotomy had a lesion with discoloration of cartilage in the medial condyle of the femur. Eighteen months after operation, five of the eight dogs that had valgus osteotomy showed fibrillation of the femoral and tibial cartilages. Mankin's scoring of the knee joint cartilages indicated statistically significant changes as compared with control subjects 7 and 18 months after surgery. Biomechanical analysis revealed shift of the mechanical axis toward the lateral compartment of the knee by the valgus osteotomy, patellofemoral malalignment, and inclination of the tibiofemoral joint line. These biomechanical alterations brought about the most severe cartilage lesions to the medial condyle of the femur and the patellofemoral joint. Cartilage fibrillation took more than 7 months to develop. Thus, this model offers a slowly progressive, well standardized, and reproducible method for the study of early changes of osteoarthritis in young beagle dogs. PMID- 9345228 TI - Displacements of the tibial tuberosity. Effects of the surgical parameters. AB - A three-dimensional computer model is used, based on the finite element method, to investigate the effects of 1-, 1.5-, and 2-cm tibial tubercle elevations and of 0.5- and 1-cm medial displacements of the tuberosity, performed with different bone shingles. Patellar kinematics and patellofemoral interface peak pressure, between 45 degrees and 135 degrees of passive knee flexion, are compared for these different surgical parameters with those of a normal knee not surgically treated. The shingle lengths of 3, 5, 7, and 10 cm have little influence on the results. Augmenting tubercle medializations decrease the lateral peak pressure but result in an overpressure of the medial facet that is 154% of the normal peak value. With knee flexion between 45 degrees and 60 degrees, increasing tubercle elevations decreases later and medial peak pressures. With flexion of more than 60 degrees, increasing elevations decrease the lateral peak pressure, but they augment and even cause overpressure on the medial facet. An overpressure on the lateral facet also is seen in midrange knee flexion (75 degrees-90 degrees) for all tubercle elevation values. Increasing tubercle elevations and medializations appear to be the predominant parameters from a biomechanical point of view. PMID- 9345229 TI - The Nicholas Andry Award-1996. The molecular pathology of osteogenesis imperfecta. AB - A systematic analysis of the molecular pathology of osteogenesis imperfecta was undertaken in 200 cases. The findings indicate that molecular defects of Type I collagen are the major cause of this disease. The mild form of osteogenesis imperfecta is caused by quantitative anomalies of Type I collagen. The other forms of the disease, which are more severe, are caused by quantitative and qualitative anomalies of Type I collagen. The mutant Type I collagen molecules are secreted poorly and are susceptible to intracellular and extracellular degradation with loss of normal and mutant collagen chains. The mutant molecules severely impair the formation of the extracellular matrix causing an abnormal architecture of dermis and bone. The molecular pathology was correlated with the clinical, radiologic, and pathologic features. As a result, the clinical classification was expanded and a new biochemical classification of osteogenesis imperfecta was developed. PMID- 9345230 TI - Back pain in an 8-year-old girl. PMID- 9345231 TI - Rehabilitation of an anterior cruciate ligament. PMID- 9345232 TI - Magnetic resonance imaging of the musculoskeletal system. Part 8. The spine, section 2. AB - Magnetic resonance imaging has revolutionized the noninvasive evaluation of degenerative disc disease and its complications. Compared with computed tomography and computed tomographic myelography, magnetic resonance allows specific determination of the nature of disc protrusions and other degenerative related soft tissues about the spine. Magnetic resonance offers the most complete evaluation of specific degenerative disorders including degenerative facet disease, spondylolysis, spondylolisthesis, spontaneous lumbar epidural hematomas, and juvenile discogenic disease. PMID- 9345233 TI - Stable vortices within vein cuffs inhibit anastomotic myointimal hyperplasia? AB - OBJECTIVES: Interposition vein cuffs improve the patency of below-knee ePTFE arterial grafts, and there is evidence that they do so, at least in part, by modifying the distribution of myointimal hyperplasia (MIH) at the distal anastomosis. Alteration of local haemodynamics is one of the mechanisms which might be involved. The purpose of this study was to characterise the local haemodynamics within an interposition vein cuff. MATERIAL AND METHODS: Flow patterns have been analysed in a laboratory model of cuffed anastomosis and compared with observations made in patients by cine intra-arterial digital subtraction angiography (IA DSA) and dynamic colour duplex scanning. RESULTS: In contrast to non-cuffed anastomoses in which the flow is predominantly laminar, cuffed anastomoses are associated with the formation of a coherent vortex. CONCLUSION: High frictional forces or shear stress exerted upon the arterial wall by the vortex could explain the beneficial effect of a cuff upon anastomotic MIH, in which case the optimal configuration of small vessel anastomoses would be that which most effectively promotes the formation of this type of vortex. PMID- 9345234 TI - Safe and cost-effective approach to carotid surgery. AB - OBJECTIVE: To evaluate the safety and cost effectiveness of carotid surgery performed altering the perioperative protocol in an attempt to decrease resource utilisation. SETTING: Department of vascular surgery in a large metropolitan teaching hospital in northern Italy. DESIGN: Prospective, non-selective study. MATERIALS AND METHODS: Three hundred and eighty carotid procedures were performed in 1995 on 343 patients (274 males, 69 females, mean age 68.2 years, range 47-86 years). The most important cost containment measures, were: (i) limiting the use of contrast arteriography to cases of dubious ultrasonographic diagnosis; (ii) routine use of loco-regional anaesthesia; (iii) postoperative admission to an intensive care unit (ICU) only in selected cases; (iv) early postoperative discharge where possible. RESULTS: Mortality was 0.26% and neurological morbidity 1.58%. General anaesthesia was required in eight patients (2.1%), and only seven patients (1.8%) were admitted postoperatively to the ICU. Arteriography was performed in 56 cases (14.7%). The average hospital stay was 5 days with a global cost of 43,036 ECU, as compared with a cost of 6764 ECU for patients treated traditionally with routine arteriography, general anaesthesia and routine ICU admission. CONCLUSIONS: Selective use of arteriography and ICU, routine use of loco-regional anaesthesia and reduced hospital stay make it possible to lower the cost of carotid surgery without sacrificing quality. PMID- 9345235 TI - Transcranial Doppler microembolus detection in the identification of patients at high risk of perioperative stroke. AB - OBJECTIVES: Perioperative ischaemic stroke is the leading cause of morbidity and mortality associated with carotid endarterectomy (CEA). The aim was to test the hypotheses that the detection of microembolic ultrasonic signals (MES) with transcranial Doppler ultrasound (TCD) during and after the operation may be of value in identifying patients at increased perioperative stroke risk. DESIGN: Open prospective case series. PATIENTS AND METHODS: Eighty-one consecutive patients undergoing CEA with TCD monitoring. Preoperative, intraoperative and interval postoperative TCD monitoring of the middle cerebral artery (MCA) ipsilateral to the operated carotid artery. On-line pre- and intraoperative MES counting and blinded off-line analysis of postoperative MES counts. End-points were any focal neurological deficit and death at 30 days postoperatively. RESULTS: MES were detected in 94% of patients intraoperatively and 71% of cases during the first postoperative hour. MES counts ranged from 0 to 25 per operative phase (range of median counts 0-8) and from 0 to 212 per hour postoperatively (range of median counts 0-4). Eight cases (10%) developed postoperative MES counts greater than 50/h. Five of these eight cases evolved ischaemic neurological deficits in the territory of the insonated MCA, indicating a strong association between frequent postoperative microembolism and the development of early cerebral ischaemia (chi 2 = 34.2, p < 0.0001). Intraoperative MES were not associated with clinical outcome measures. CONCLUSIONS: MES counts of greater than 50/h in the early postoperative phase of carotid endarterectomy are predictive of the development of ipsilateral focal cerebral ischaemia. PMID- 9345236 TI - Antibodies to cardiolipin may increase the risk of failure of peripheral vein bypasses. AB - OBJECTIVES: To assess the association between antibodies to cardiolipin and infrainguinal vein graft patency. MATERIALS AND METHODS: Plasma levels of antibodies to cardiolipin, haemostatic factors, lipids and the smoking marker carboxyhaemoglobin were determined preoperatively and 6 weeks postoperatively in 80 patients undergoing infrainguinal vein bypass surgery. Bypass patency was assessed by ankle blood pressure measurements and ultrasound duplex scanning at 1 week, 6 weeks, 3, 6, 9 and 12 months. A localised increase in the graft peak systolic velocity by a factor of 2.5 or more was considered to indicate a significant stenosis. RESULTS: Antibodies to cardiolipin were identified in seven (9%) patients preoperatively. In four of these seven patients the bypasses thrombosed within 3 months after surgery and another two developed stenoses. At 6 months the primary bypass patency, i.e. patency without stenosis, was 14% (95% confidence interval (CI) 0-33%) in patients with antibodies to cardiolipin, as opposed to 57% (95% CI 45-69%) in patients without these antibodies (log rank test: p = 0.03). Diabetes mellitus was also associated with a reduced 6 months primary bypass patency (38% (95% CI 16-60%) vs. 58% (95% CI 45-71%), p = 0.006). A Cox regression analysis showed that both the presence of antibodies to cardiolipin and diabetes independently contributed towards predicting the overall risk of bypass failure. CONCLUSION: Antibodies to cardiolipin were identified in 9% of patients undergoing infrainguinal vein bypass surgery and appeared to be associated with increased risk of bypass failure. PMID- 9345237 TI - A review of the management of abdominal aortic aneurysms in patients following cardiac transplantation. AB - OBJECTIVES: To describe the presentation, preoperative assessment and postoperative management of patients presenting with an infrarenal abdominal aortic aneurysm following a previous cardiac transplant. METHODS: The case histories of three patients have been examined and a literature review performed. CONCLUSIONS: The majority of patients developing aortic aneurysms had undergone cardiac transplantation for ischaemic cardiomyopathy and thus require detailed assessment of cardiac function preoperatively to exclude accelerated coronary artery disease in the graft. Full invasive cardiac monitoring during surgery is mandatory to maintain haemodynamic stability in patients with a dennervated heart and to avoid postoperative renal failure. The higher incidence of pulmonary and wound complications are discussed, together with protocols for maintaining adequate immunosuppression. Finally, data supporting a higher prevalence and more rapid expansion of aortic aneurysms in immunosuppressed patients is considered and evidence-based recommendations made regarding aneurysm screening in these patients. PMID- 9345238 TI - A paper for debate: vein versus PTFE for critical limb ischaemia--an unfair comparison? AB - INTRODUCTION: There is a widely held view that vein grafts for infrainguinal arterial reconstruction perform much better than prosthetic conduits, the best of which seems to be PTFE. Many randomised studies have been conducted which confirm this opinion, but is the difference as large as it is thought to be? One interesting feature of published trials is that the results for obligatory PTFE (when no vein is available) were much worse than the results for randomised PTFE grafts. The only way to explain this is that these groups of patients were not similar, and there are probably other factors which contribute to the difference in results when vein and PTFE grafts are compared. MATERIALS AND METHODS: A consecutive series of 109 femoro-infrapopliteal grafts undertaken for critical limb ischaemia was analysed to see the difference between vein and PTFE with vein cuff grafts. RESULTS: Vein grafts were superior to PTFE grafts when the whole cohort was included (p = 0.0038); however, there was no significant difference when the patients were stratified for inflow and runoff status. CONCLUSIONS: The difference between vein and PTFE has probably been exaggerated in the past, due to differences in risk factors and in the extent of arterial disease between the two groups of patients. The advantage of vein becomes more significant with time. PMID- 9345239 TI - Post-ischaemic organ dysfunction: a review. AB - OBJECTIVES: The aim of this review is to consider the pathophysiology of ischaemia-reperfusion in organs that may be affected by either its local or remote consequences. Potential therapeutic strategies are also considered. DESIGN: A general discussion of the biochemical (including oxygen free radicals, complement, cytokines) and cellular events (endothelial cells, neutrophils) responsible for the mediation of reperfusion injury is presented, with special consideration of the organ-specific differences affecting the myocardium, central nervous system, gut, liver, kidney and skeletal muscle. Similarly, events which promote remote organ injury are described. CONCLUSIONS: Although it is recognised that prolonged ischaemia results in tissue and organ damage, the concept of reperfusion-induced tissue injury, defined as tissue damage occurring as a direct consequence of revascularisation, is relatively recent. Such events may increase the morbidity and mortality of patients undergoing vascular reconstruction, trauma surgery and transplantation. A clear understanding of the factors responsible for its development is therefore vital if protocols that reduce its impact are to be developed. PMID- 9345240 TI - Transfemoral treatment for iliac occlusive disease with endoluminal stent-grafts. AB - OBJECTIVES: Percutaneous treatment of iliac artery occlusive disease has replaced open vascular reconstruction for several indications. A balloon angioplasty with or without stent is not an option in the presence of infrainguinal extension of the disease. The authors describe a technique that allows the construction of an aorto- or iliofemoral graft through a single groin incision, using a 4 mm PTFE graft, anchoring it proximally with a Palmaz stent and dilating both to the desired diameter. DESIGN: Retrospective non-randomised study. MATERIALS AND METHODS: Nineteen procedures were performed in 16 patients mainly because of ischaemic rest pain, often with trophic skin changes or minor gangrene. Three patients had a bilateral procedure. Twelve patients had one or more associated procedures: 10 distal bypasses, one thrombectomy, one reimplantation of a distal bypass on the iliofemoral graft, one contralateral profundaplasty and two stents of the contralateral common iliac artery. RESULTS: Two patients died, one of small bowel ischaemia and the other of a myocardial infarction. During the mean follow-up of 8.8 months, two graft thromboses occurred. In another patient bilateral stenting of a residual stenosis was necessary. CONCLUSIONS: Our experience shows that the reported technique is feasible. Whether the procedure is truly "less invasive" and the long-term results acceptable remains to be shown. PMID- 9345241 TI - Glutaraldehyde-tanned bovine carotid artery graft for infrainguinal vascular reconstruction: 5-year follow-up. AB - OBJECTIVE: To assess the long-term patency of a modified biological conduit, the glutaraldehyde-tanned bovine carotid artery, in above-knee infrainguinal arterial reconstruction. PATIENTS AND METHODS: Prospective follow-up of a cohort of 58 above-knee femoropopliteal grafts in 55 patients. Graft patency was assessed at yearly intervals with doppler ankle pressure measurements. RESULTS: The median follow-up period has been 67 months. Nine grafts occluded within 30 days of surgery and a further 19 graft closures have been observed in the follow-up period. The overall cumulative primary graft patency at 1, 3 and 5 years was 70%, 61% and 56%, respectively. If the 30-day graft failures are excluded, the primary graft patency rises to 83%, 74% and 68% at 1, 3 and 5 years, respectively. Six limbs have been amputated, four above the knee and two below the knee. There were no graft aneurysms and no graft infections. CONCLUSION: Results indicate that the modified bovine carotid artery graft with an above-knee anastomosis does not seem to be inferior to PTFE, but is inferior to reversed vein. Modified biological conduits offer a reasonable alternative to synthetic grafts for infrainguinal arterial reconstruction and appear to maintain acceptable long-term mechanical stability. PMID- 9345242 TI - Subintimal angioplasty of infrapopliteal occlusions in critically ischaemic limbs. AB - OBJECTIVE: To review the outcome of subintimal angioplasty of infrapopliteal artery occlusions in critically ischaemic limbs. DESIGN: Retrospective review. MATERIALS: Twenty-eight consecutive limbs with critical ischaemia that had undergone subintimal angioplasty of infrapopliteal occlusions. RESULTS: There were 32 infrapopliteal artery occlusions in 28 critically ischaemic limbs in 27 patients. The median (range) patient age was 81 (48-88) years. Seventeen limbs (61%) were ulcerated, seven (25%) were gangrenous and four (14%) had rest pain only. Twenty-five (89%) procedures were to a single calf vessel, and three (11%) procedures were to multiple calf vessels. The median (range) length of the occlusions was 7 (2-30) cm. The initial technical success rate was 27/32 (84%). There were three minor complications--one groin haematoma, one vessel perforation and one distal embolus. There were no limbs lost as a result of the procedure itself and the 30-day mortality was zero. The 12-month actuarial haemodynamic and symptomatic patencies (including initial failures) were 53% and 56%, respectively. The 12-month limb salvage rate was 85% and patient survival was 81%. CONCLUSION: We conclude that subintimal angioplasty in patients with infrapopliteal artery occlusions and critical ischaemia is safe, effective, and offers a low-risk alternative to distal reconstructive surgery. PMID- 9345243 TI - Clostridium difficile colitis after aortic surgery. AB - OBJECTIVE: To determine the incidence and outcome of Clostridium difficile colitis (CDC) following aortic surgery. DESIGN: Retrospective clinical study, and case-note review. PATIENTS: Of 180 patients undergoing aortic surgery for either aneurysmal or occlusive disease between 1 September 1994 and 31 August 1996 (24 months), 15 (8.4%) developed CDC. There were 12 male and three female patients of median age 65 (range 46-84). RESULTS: Two patients died from multiple organ failure in association with CDC, one of whom underwent negative relaparotomy for suspected ischaemic bowel because the diagnosis of CDC had not been entertained. Previously identified risk factors for CDC comprised: age > 65 (eight); renal impairment (four); chronic obstructive airways disease (seven); coexistent malignancy (three); admission from another hospital (four); H2 antagonist therapy (13); ITU (nine); and/or HDU care (14). Diarrhoea commenced a median of 9 (range 5-26) days, and CDC, was diagnosed a median of 14 (range 10-26) days after operation. All patients received intravenous Cefuroxime, originally prescribed as prophylaxis, for a median of 6 (range 3-16) days prior to onset of CDC. Two patients received 1 additional antibiotic; one received 2; two received 3; and one received 4 prior to onset of CDC. CONCLUSIONS: CDC is a common and potentially serious complication of vascular, and in particular, aortic surgery. Although such patients often possess several risk factors for CDC, colitis frequently follows prolonged 'prophylactic' cephalosporin administration, which should therefore be avoided. PMID- 9345244 TI - Solitary intrarenal aneurysm. PMID- 9345245 TI - Surgical approach in the treatment of arterial aneurysms associated with Behcet's disease. PMID- 9345246 TI - Endovascular management of a non-penetrating traumatic axillary artery occlusion. PMID- 9345247 TI - Arterio-venous fistula following above-knee amputation. PMID- 9345248 TI - Understanding shame in adults: retrospective perceptions of parental-bonding during childhood. AB - The association between perceptions of parental-bonding style during childhood and moral affect of shame at young adulthood were examined with 264 women and 140 men (mean age [+/- SD] = 20.4 +/- 1.6 years old). Shame affect was significantly positively related to fear of negative evaluation by others and social avoidance, and negatively related to recalled parental care in one's childhood. Multiple regression analyses indicated that maternal protectiveness, paternal care, fear of negative social evaluation, and social avoidance were significant predictors of shame, explaining 41% of the variance. Results support object relations theory, which states that shame is a moral affect associated with social evaluation apprehension and may have developmental implications for one's parental relations. PMID- 9345249 TI - The effects of subliminal symbiotic stimulation on free-response and self-report mood. AB - Research has shown that subliminal presentation of MOMMY AND I ARE ONE (MIO) can help improve adaptive functioning. Two experiments tried to determine whether changes in mood, especially free-response mood, could help explain these findings. In one experiment, 20 men were randomly assigned to receive either a subliminal MIO or control stimulus. Results showed predicted effects on a free response and no effects on a self-report mood measure. In the other experiment, 54 male subjects randomly received one of three subliminal stimuli. They evidenced the same pattern of mood results. Sentential semantics were shown to be relevant to the obtained results. Ascending threshold and 150 forced-choice discrimination trials demonstrated that subjects could not report stimulus content. It was concluded that MIO effects were attributable to unconscious processing of the entire message and that free-response mood may partly mediate these effects. Suggestions for future research were offered. PMID- 9345250 TI - Latent variable models of functional somatic distress. AB - Latent variable models of functional somatic symptoms were estimated for a sample of 686 family medicine patients. Symptom items from the NIMH Diagnostic Interview Schedule were selected to approximate diagnoses of fibromyalgia syndrome (FMS), chronic fatigue syndrome (CFS), and irritable bowel syndrome (IBS). Confirmatory factor analysis demonstrated that hypothesized latent variables of somatic depression, somatic anxiety, FM-like, CF-like, and IB-like syndromes fit the observed covariations better than models hypothesizing fewer latent variables. Results offer tentative confirmation of functional somatic syndromes as discrete entities and suggest that relaxing the diagnostic criteria for somatization may identify individuals with distress limited to a single functional system. PMID- 9345251 TI - Negative symptoms in stroke patients and length of hospital stay. AB - The purpose of this study was to assess whether the presence and severity of psychiatric symptoms in stroke patients correlate with their length of stay (LOS) in a rehabilitation unit, with special emphasis on the role of negative symptoms (NS). Twenty-three stroke patients, consecutively recruited from the inpatient rehabilitation unit, were evaluated on admission with the Mini-Mental State Examination (MMSE), the Positive and Negative Symptom Scale (PANSS), the Hamilton Depression Rating Scale (HDRS), the Scale for Assessment of Negative Symptoms (SANS), and the Functional Independent Measure (FIM). NS scores significantly correlated with LOS, with SANS total score being the most informative, and the attentional impairment subscale the least. The group of patients with pronounced NS stayed in the hospital twice as long as patients with the score on the NS subscale of PANSS below 16. These two groups did not differ in their cognitive performance or in the positive symptom subscale of PANSS scores. Total FIM score on admission was lower and HDRS scores higher in patients with pronounced NS. However, these differences, unlike those of LOS, have not reached statistical significance. The presence and severity of NS in stroke patients are associated with a longer hospital stay. Identification and treatment of NS might lead to a faster discharge from rehabilitation unit. PMID- 9345252 TI - Expectations and motives for alcohol use in a psychiatric outpatient population. AB - The purpose of the study was to assess alcohol expectancies and motives of psychiatric outpatients with and without comorbid current or lifetime substance use disorders. Seventy-five psychiatric outpatients with diagnoses of mood disorders, anxiety disorders, and substance use disorders were administered the Alcohol Effect Expectancy Questionnaire-Abridged Version and the Drinking Motives Measure. Results demonstrated that the internal reliabilities for the two scales were comparable with those reported for these measures in the general population. Psychiatric outpatients with a history of comorbid substance use disorders reported greater expectancies and motives for using alcohol than did patients with no such history. In addition patients with comorbid alcohol and drug use disorders, and only comorbid alcohol use disorder, showed significantly greater expectancies and motives for alcohol use than patients with only comorbid drug use disorders and patients with no history of comorbid substance use disorder. We discuss the implications of the findings for role of expectancies and motives in the maintenance and treatment of substance abuse in psychiatric patients. PMID- 9345253 TI - Substance use disorders in a geriatric psychiatry outpatient clinic: prevalence and epidemiologic characteristics. AB - This study was conducted to determine the prevalence of substance use disorders in a geriatric psychiatry outpatient clinic. The overall prevalence for any substance use disorder was 20% (N = 28). The prevalence of benzodiazepine dependence was 11.4% (N = 16); the prevalence of alcohol dependence was 8.6% (N = 12); and the prevalence of prescription narcotic dependence was 1.4% (N = 2). These findings suggest that substance use disorders in the geriatric psychiatry outpatient population exist to a significantly greater extent than previously reported. Descriptive statistics were used to characterize patients with benzodiazepine dependence, alcohol dependence, and no substance use disorder. These groups were compared on demographic and clinical variables using one-way analysis of variance (ANOVA) and chi-squared statistical techniques. Clinicians working in comparable outpatient settings may be in a better position to prevent, detect, and treat substance use disorders in their patients as a result of increased awareness of its epidemiologic characteristics in this population. PMID- 9345254 TI - Individual and community-level variation in intensity and diversity of service utilization by homeless persons with serious mental illness. AB - This study examines individual client- and community-level sources of variation in service use among clients entering 18 community treatment programs for homeless mentally ill persons as part of a national demonstration project. Assessment data on 1,828 clients were used to evaluate the relationship of a) individual client characteristics and b) site of entry, to both the intensity and diversity of service use. Hierarchical multiple regression was used to identify the relative importance of client characteristics and site of entry. Client characteristics explained only 2% to 3% of the variance in service use. Inter site variation accounted for 2 to 3 times as much of the variance. Inter-site differences account for substantially more of the variance in service use among homeless persons with mental illness than individual client characteristics. Further studies are needed to identify specific community-level factors that account for these variations. PMID- 9345255 TI - Failure of fusion of the septum pellucidum and the heterogeneity of schizophrenia. PMID- 9345256 TI - A comparison of neuropsychological deficits in familial schizophrenics, nonfamilial schizophrenics, and normal controls. PMID- 9345257 TI - Is positive placebo response in chronic schizophrenia investigator-dependent? PMID- 9345258 TI - Lifetime prevalences of nine common psychiatric/personality disorders in female domestic abuse survivors. PMID- 9345259 TI - Phospholamban: a protein coming of age. AB - Phospholamban is a major regulator of the kinetics of cardiac contractility, through its ability to regulate the function of the cardiac SR Ca(2+)-pump and thus the SR Ca2+ load. In vitro expression studies have provided significant information on the structure/function of the phospholamban/Ca(2+)-pump interaction. Furthermore, the generation of genetic animal models with altered phospholamban expression levels have permitted a through understanding of the physiological role of this regulatory phosphoprotein. Future studies aimed towards crystallization of phospholamban and the SR Ca(2+)-ATPase in their native SR environment may provide clues to their tertiary and quaternary structures and may further elucidate the mechanisms underlying the phospholamban regulatory effects in vivo. PMID- 9345260 TI - Interleukin-15 stimulates C2 skeletal myoblast differentiation. AB - Interleukin-15 (IL-15) is a cytokine which is highly expressed in skeletal muscle, and which stimulates muscle protein accretion in cultured skeletal muscle fibers. Using parental C2 skeletal myoblasts, no significant effects of IL-15 on skeletal muscle differentiation were observed. To test the hypothesis that IL-15 may stimulate skeletal muscle differentiation if the strong differentiation inducing effects of autocrine insulin-like growth factor (IGF) production were inhibited, a C2 myoblast subline (C2-pBP4) was stably transfected with an expression vector for rat IGF binding protein-4 (IGFBP-4). Differentiation responses to autocrine and exogenous IGFs in C2-BP4 myoblasts were reduced 3- to 4-fold in C2-BP4 cultures compared to C2-pLXSN cultures, a subline transfected with a control plasmid. Addition of IL-15 to C2-pBP4 myoblasts doubled the number of differentiated muscle cells which arose. These findings indicate that IL-15 can stimulate myogenic differentiation in conditions in which the strongly differentiative effects of the IGFs are inhibited. The differentiative activity of IL-15 may be of physiological significance in conditions in which IGF concentrations are low or in which the IGFs are sequestered by binding proteins. PMID- 9345261 TI - The CD40 ligand directly activates T-lymphocytes via tyrosine phosphorylation dependent PKC activation. AB - The activation of B-lymphocytes depends critically on the interaction of the CD40 receptor with its ligand. Here, we provide evidence that the CD40 ligand (CD40L) also functions as a direct stimulatory molecule for T-lymphocytes. Activation of T-lymphocytes via CD40L induces tyrosine phosphorylation of cellular proteins including PLC gamma. Tyrosine phosphorylation of PLC gamma correlates with an IP3 and Ca(2+)-release and an activation of PKC. Inhibition of src-like tyrosine kinases by Herbimycin A prevents these activation events suggesting a crucial role of tyrosine phosphorylation in T-lymphocyte activation via CD40L. PMID- 9345262 TI - Purification and partial amino acid sequence of a novel protein of the reticulocalbin family. AB - Binding proteins in neuronal membranes for a phospholipase A2 with presynaptic neurotoxicity have been purified. Three polypeptides of 87, 65, and 50 K Da were obtained from the synaptic membrane fraction of guinea pig brain utilizing an immobilized crotoxin (a phospholipase A2) column. For large scale purification, porcine brain was used instead, and two polypeptides of 50 and 18 K Da were found. The 65 and 18 K polypeptides may represent hitherto unidentified components of the crotoxin-binding proteins. Partial N-terminal amino acid sequence and a partial sequence for an internal peptide fragment have been determined for the 50 K polypeptide. Search of protein data bank reveals that this polypeptide or protein is a novel member of the reticulocalbin family of calcium-binding proteins. PMID- 9345264 TI - Generation and characterization of mice lacking gastrin-releasing peptide receptor. AB - Gastrin-releasing peptide (GRP) is a mammalian bombesin-like peptide which is widely distributed in the central nervous system as well as in the gastrointestinal tract. GRP binds to its high affinity receptor (GRPR) to elicit a wide spectrum of biological effects on behavior, digestion, and metabolism. To define the in vivo function of GRPR, we generated GRPR null mutant mice by gene targeting. The intracerebroventricular administration of GRP caused hypothermia in wild-type mice, but not in mutant mice. The GRPR deficient mice showed significantly increased locomotor activity during the dark period, and social responses scored by sniffing, mounting, and approaching behaviors against an intruder. Aggressive scores such as fighting and biting were not altered in the mutant mice. These phenotypes were observed in mice generated from two independent ES cell clones and backcrossed to a C57BL/6J background. The GRPR deficient mice should be useful for studying the bombesin system in vivo. PMID- 9345263 TI - Platelet-activating factor stimulates calcium-dependent activation of protein tyrosine kinase Syk in a human B cell line. AB - In ASK.0 B lymphoblastoid cells, platelet activating factor (PAF) induced a rapid increase in Syk protein-tyrosine kinase activity which was insensitive to pertussis toxin (PTX) but was abolished by the phopholipase C inhibitor, U73122. In parallel, PAF-induced Ca2+ mobilization was also insensitive to PTX and was almost completely inhibited by U73122. Incubation of ASK.0 cells with the compounds that increase intracellular Ca2+ (i.e., the ionophore A23187, thapsigargin which releases Ca2+ from internal store) mimicked the effect of PAF on Syk kinase activity. Loading cells with the intracellular Ca2+ chelator, bis (O-aminophenoxy)-ethane-N,N,N',N'-tetraacetoxymethyl ester (BAPTAAM), completely inhibited the activation of Syk kinase in response to PAF, thapsigargin and ionophore. These results suggest that intracellular free Ca2+ seems to be critical for PAF-induced activation of Syk kinase in human B lymphoblastoid cells. PMID- 9345265 TI - Promoter characterization of the rat Na+/I- symporter gene. AB - The Na+/I- symporter is the molecule that mediates active iodide uptake in the thyroid gland. A 16.4 kb genomic DNA fragment of the rat Na+/I- symporter gene (rNIS) was isolated, restriction mapped and a 2 kb region immediate 5' to the ATG site was sequenced. The transcription start site for rNIS was mapped to -98 nucleotide (nt) relative to the ATG translation initiation site. A series of 5' genomic DNA fragments ranging from 233 bp to 8 kb were tested for promoter activity in both thyroid and non-thyroid cells. Our results indicate that the DNA regulatory elements within 8 kb of the 5' flanking region of rNIS are not sufficient to confer thyroid-selective transcription. However, consistent with the clinical observation that NIS expression is reduced in thyroid tumors, the rNIS promoter activity is suppressed in ret/PTC1 transformed NIH3T3 cells, compared to non-transformed NIH3T3 cells. PMID- 9345266 TI - GTP loading of farnesylated p21Ras by insulin at the plasma membrane. AB - Insulin promotes the phosphorylation and activation of farnesyltransferase (FTase) in a time- and a dose-dependent manner. Increased FTase activity results in a larger pool of farnesylated p21Ras and allows for enhanced GTP loading. Insulin significantly increases the pool of farnesylated p21Ras from 20-25% in quiescent 3T3-L1 fibroblasts to approximately 70%, most of which is targeted to the plasma membrane. Furthermore, insulin promotes GTP loading of plasma membrane and not cytosolic p21Ras. The half-life of plasma membrane-associated farnesylated p21Ras is approximately 6 hours, and is identical in control and insulin-treated cells. We have also observed a direct correlation between the amounts of farnesylated p21Ras at the plasma membrane and the magnitude of insulin-induced GTP loading of p21Ras. PMID- 9345267 TI - Stationary phase-specific mRNAs in Escherichia coli are polyadenylated. AB - Polyadenylation of Escherichia coli specific mRNAs has so far been studied primarily during the exponential phase of growth. As part of an investigation of the polyadenylation of E. coli mRNAs in different physiological contexts, we studied mRNA polyadenylation in stationary phase by preparing a cDNA library from stationary phase RNA using oligodeoxythymidylate primers and analyzing the nucleotide sequence of cDNA clones corresponding to the stationary phase-specific genes, rpoS, bo1A, and dps. The sites of polyadenylation were found to be primarily in the 3'-untranslated region, either at the putative rho-independent transcription termination site (dps) or at several different sites upstream of the putative rho-independent terminator. A few examples of polyadenylation within the coding regions were also found, suggesting that nucleolytic degradation often preceded polyadenylation. In contrast to the poly(A) tracts characteristic of exponentially growing cells, many of the uncoded poly(A) tracts associated with stationary phase mRNA were interspersed with other nucleotide residues. The observation of post-transcriptional polyadenylation of specific stationary phase mRNAs in E. coli, some of which are transcribed by the RNA polymerase associated with sigma, demonstrates that mRNA polyadenylation is not confined to the exponential phase of growth. PMID- 9345268 TI - Posttranscriptional regulation of MRP/GS-X pump and gamma-glutamylcysteine synthetase expression by 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2 chloroethyl)-3-nitrosourea and by cycloheximide in human glioma cells. AB - Treatment of human glioma A172 cells with 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethy-3-nitrosourea (ACNU) for 2 to 4 hr resulted in a 2- to 3 fold increase in steady-state levels of multidrug resistance-associated protein (MRP) and gamma-glutamylcysteine synthetase (gamma-GCS) mRNA. Nuclear run-on assays revealed a less than 0.5-fold increase in transcription rates of these genes under the same treatment conditions, suggesting that posttranscriptional regulation plays an important role for the increased mRNA levels. In the absence of ACNU, rates of MRP and gamma-GCS mRNA degradation were similar in A172 cells as determined by incubating cells with the RNase inhibitor, Actinomycin D. ACNU treatments resulted in increased MRP mRNA stability. Induction of MRP and gamma GCS mRNA by ACNU apparently did not require de novo protein synthesis as determined by the use of protein synthesis inhibitor cycloheximide (CHX). However, CHX alone could induce accumulation of gamma-GCS mRNA, also by posttranscriptional mechanism. Taken together, these results demonstrate that (i) posttranscriptional regulation is primarily involved in the induction of MRP and gamma-GCS expression by ACNU and CHX in human glioma cells; and (ii) despite the fact that these two genes have been reported to be frequently co-expressed, their responses to the treatments of RNA and protein synthesis inhibitors are not the same. PMID- 9345269 TI - Comparison of exocytotic mechanisms between acetylcholine- and catecholamine containing vesicles in rat pheochromocytoma cells. AB - The molecular mechanisms of exocytosis from two types of secretory organelles, synaptic-like microvesicles and secretory vesicles, were compared by measuring acetylcholine (ACh) and catecholamine (CA) release from a newly isolated PC12 subclone, PC12-C3 which contains a high level of Ach. Digitonin-permeabilized PC12-C3 cells released both transmitters with similar Ca(2+)-dependency. Ca(2+) evoked Ach and CA release from permeabilized cells were increased in the presence of MgATP, suggesting the existence of a MgATP-dependent priming step prior to the Ca(2+)-triggered fusion step in both ACh release and CA release. The non hydrolyzable analogue of GTP guanosine 5'-(gamma-thio)triphosphate (GTP gamma S), produced both ACh and CA release from permeabilized cells in the absence of Ca2+. Pretreatment with a phorbol ester which activates protein kinase C, potentiated depolarization-induced ACh and CA release from unpermeabilized cells. These results indicated that exocytosis from two distinct vesicle populations are mediated by the same basic molecular mechanisms. PMID- 9345270 TI - Angiotensin II injection into mice increases the uptake of oxidized LDL by their macrophages via a proteoglycan-mediated pathway. AB - Angiotensin II (Ang-II) has been shown to possess several atherogenic properties including its ability to induce macrophage-mediated oxidation of LDL and to form Ang-II-modified LDL which is taken up by macrophages at enhanced rate. Oxidized LDL (Ox-LDL) is also taken up by macrophages at enhanced rate via several scavenger receptors, leading to macrophage cholesterol accumulation. In the present study we examined the effect of Ang-II on the uptake of Ox-LDL by peritoneal macrophages derived from Balb/c mice (MPM). Intraperitoneal injection of Ang-II (10(-7) M, once daily for a period of 2 days) to the mice resulted in an increased Ox-LDL uptake up to 60%, in comparison to macrophages from placebo treated mice. Similar results were obtained when Ang-II (10(-7) M) was injected to the mice twice a week for a period of three months. This Ox-LDL uptake was Ang II dose-dependent. The cellular uptake of acetylated-LDL (Ac-LDL), another ligand for scavenger receptors, however, was not affected by Ang-II injection to the mice. Furthermore, preincubation of the MPM with the monoclonal antibody, anti CD36, reduced macrophage uptake of Ox-LDL in Ang-II-treated mice by only 11%. Ang II administration to mice resulted in a 60% increase in the macrophage cellular proteoglycan content. Chondroitinase treatment of MPM decreased Ox-LDL cellular uptake by 20% and by 38% in placebo-treated and Ang-II-treated cells, respectively. We thus conclude that Ang-II administration to mice enhances their macrophage Ox-LDL uptake via its stimulating effect on cellular proteoglycan content and this process can lead to foam cell formation and atherosclerosis. PMID- 9345271 TI - Stimulation of TK1 lymphoma cells via alpha 4 beta 7 integrin results in activation of src-tyrosine- and MAP-kinases. AB - The lymphocyte integrin alpha 4 beta 7 is a cell surface adhesion receptor involved in initiating lymphocyte homing to gut-associated/mucosal lymphoid tissues by binding the mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Other known ligands are vascular cell adhesion molecule-1, fibronectin, and the alpha 4 integrin chain itself. Here, we demonstrate that stimulation of the alpha 4 beta 7 integrin through its alpha 4 subunit (mAb R1-2), beta 7 subunit (mAb M293), or the combinatory epitope (mAb DATK32) enhances tyrosine phosphorylation of several cellular proteins in the murine TK1 lymphoma cell line. The two src kinases p56lck and p59fyn were identified as possible mediators and substrates of the detected tyrosine phosphorylation. Furthermore, we observed activation of the MAP-kinases ERK1/2. PMID- 9345272 TI - Gene cloning and characterization of maleate cis-trans isomerase from Alcaligenes faecalis. AB - Maleate cis-trans isomerase, which catalyses the conversion of maleate to fumarate, was purified and characterized from Alcaligenes faecalis IFO13111. The molecular weight of maleate isomerase was estimated as 60 kDa, consisting of a 28 kDa dimer as shown by gel-filtration chromatography and SDS-PAGE analysis. Kinetic studies showed that the Michaelis constant for maleate was 4.0 x 10(-5) M. The reverse reaction (fumarate to maleate) activity of the enzyme was detected even though it was quite weak. The maleate isomerase gene (maiA) was cloned by hybridization using the oligonucleotide DNA probes designed on the basis of the determined N-terminal amino acid sequences of the purified enzyme. The determined DNA sequence of the maiA gene contains an open reading frame which encodes a 254 amino-acid sequence. The amino acid sequence of the maiA gene product shows no significant homology to any amino acid sequences in the protein data base. PMID- 9345273 TI - Decreased retinoylation in NIH 3T3 cells transformed with activated Ha-ras. AB - Retinoylation (retinoic acid acylation) is a post-translation modification of proteins occurring in a variety of mammalian cell lines and in vivo. To gain further knowledge of the role of retinoylation we studied it in NIH 3T3 cells and NIH 3T3 cells transformed by an activated Ha-ras oncogene (NIH Ha-ras-3T3 cells). In serum-free medium retinoic acid (RA) inhibited growth of NIH 3T3 cells but did not inhibit growth of NIH Ha-ras-3T3 cells. After incubation with [3H]RA, the level of retinoylated protein in NIH 3T3 cells was about 1.5-fold greater than in NIH Ha-ras-3T3 cells. On one-dimensional polyacrylamide gel electrophoresis, both the rate and the extent of retinoylation were greater in NIH 3T3 cells. We detected about 40 retinoylated proteins in NIH 3T3 cells by two-dimensional polyacrylamide gel electrophoresis. Only about 15 proteins were retinoylated, but at reduced levels, in NIH Ha-ras-3T3 cells. These results suggest that the activated ras oncogene inhibits retinoylation. This inhibition may in turn be related to the loss of other RA responses of NIH 3T3 cells, including growth inhibition, retinoic acid catabolism, down-regulation of fibronectin biosynthesis, and induction of tissue-type transglutaminase, which are not seen to the same extent in NIH Ha-ras-3T3 cells. PMID- 9345274 TI - Carotenoid pigments of an antarctic psychrotrophic bacterium Micrococcus roseus: temperature dependent biosynthesis, structure, and interaction with synthetic membranes. AB - Pigmentation in a psychrotrophic M.roseus was found to be increased when the bacteria were grown at 5 degrees C as compared to its pigmentation at 25 degrees C. In addition more polar pigments were synthesised at low temperature. The pigments were identified as bacterioruberins and were demonstrated to bind to synthetic membranes of phosphatidylcholine with almost equal affinity, irrespective of the polarity of the pigments. PMID- 9345275 TI - Glycated albumin stimulates fibronectin and collagen IV production by glomerular endothelial cells under normoglycemic conditions. AB - Albumin modified by Amadori glucose adducts, formed in increased amounts in diabetes, stimulates the synthesis of matrix by renal glomerular mesangial cells and has been causally linked to the pathogenesis of diabetic nephropathy. However, the effect of glycated albumin on the biology of glomerular endothelial cells, which elaborate a basement membrane that undergoes thickening in diabetes, has not been investigated. We used well-characterized rat glomerular endothelial cells to examine the influence of glycated albumin on the synthesis of extracellular matrix proteins by these cells in culture. Concentrations of glycated albumin that are present in clinical specimens stimulate fibronectin and collagen IV production by glomerular endothelial cells, and this effect is operative under normoglycemic conditions. These results support the hypothesis that increased glycated albumin contributes to glomerular basement membrane thickening in diabetes. PMID- 9345276 TI - Mutations of the p53 gene in human lung cancer from chromate-exposed workers. AB - We examined p53 mutations in 20 cancer samples from 19 chromate workers with lung cancer by Polymerase chain reaction-Single strand conformation polymorphism analysis and direct sequencing. Six missense mutations were identified in 4 (20%) of the 20 chromate lung cancer samples. Fewer mutations were found in the patients with lung cancers who had been exposed to chromate than in those who had not. However, the pattern of p53 mutations in lung cancer patients exposed to chromate differed from that of common lung cancers in 3 respects. There were no apparent G to T transversions, which are common base changes in lung cancers. Half of the mutational sites (3/ 6) had changes of AT base-pairs, and 2 of 4 mutational tumor samples had double missense mutations. Our results suggested that chromate exposure may induce point mutation of the p53 gene. PMID- 9345277 TI - The release mechanism of platelet-activating factor during shear-stress induced platelet aggregation. AB - We previously reported that 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet-activating factor, PAF), released from activated platelets stimulated with thrombin plus collagen, is associated with platelet microparticles. In the present study, we found that PAF is concentrated and associated with microparticles released from high shear stress-induced activated platelets. The total amount of PAF released from 3 x 10(8) platelets under high shear stress (108 dyne/cm2) was 3.2 +/- 0.6 x 10(-15)mol (n = 5, mean +/- S.D.). Eighty percent of the PAF released from the platelets was recovered in the microparticle fraction after ultracentrifugation in the presence of albumin. Under high shear stress, PAF was not released from platelets within 3 minutes, although microparticles were released. In conclusion, microparticles released from activated platelets in the rheological condition of high shear stress are major carriers of PAF. PMID- 9345278 TI - Phenotypic consequences after restoration of lymphopenia in the diabetes-prone BB/OK rat. AB - Besides other factors, lymphopenia (ll) is essential for development of insulin dependent diabetes mellitus in the BB rat. It is unknown which phenotypic consequence may have a non-lymphopenia. Therefore, the region containing the lymphopenia gene of the BB/OK rat was replaced with that of the non-lymphopenic (LL) and hypertensive rat, SHR (11 cM, D4Mit6-LL-Npy-Spr). The resulting congenic strain, BB.LL, did not develop diabetes up to an age of 30 weeks and was non-T lymphopenic. The SHR region did not influence the blood pressure of the BB.LL rat, but influenced obviously the motor activity, body weight, serum lipids, and the 24h urine excretion of urea, sodium, and potassium. PMID- 9345279 TI - Advanced glycation endproducts stimulate mitogen-activated protein kinase and proliferation in rabbit vascular smooth muscle cells. AB - Advanced glycation end products of bovine serum albumin (AGEs-BSA) exhibited biphasic effects on the proliferation of cultured rabbit vascular smooth muscle cells (VSMCs) in terms of [3H]thymidine incorporation and cell number count; a stimulatory effect was observed at 1-10 micrograms/ml and an inhibitory effect at more than 20 micrograms/ml, while it inhibited [3H]thymidine incorporation even at 1-10 micrograms/ml in cultured bovine vascular endothelial cells (VECs). Transient activation of p42 mitogen-activated protein kinase (MAPK) with a peak at around 5 min and a subsequent sustained phase was induced by AGEs-BSA in VSMCs, but not in VECs. The dependence of MAPK activation on AGEs-BSA dose was correlated with that of VSMCs proliferation. PMID- 9345280 TI - Isolation and characterization of the yeast mRNA capping enzyme beta subunit gene encoding RNA 5'-triphosphatase, which is essential for cell viability. AB - The yeast Saccharomyces cerevisiae mRNA capping enzyme is composed of two subunits of alpha (52 kDa, mRNA guanylyltransferase) and beta (80 kDa, RNA 5' triphosphatase). We have isolated the alpha subunit gene (CEG1) by immunological screening. In this report, with the aid of partial amino acid sequences of purified yeast capping enzyme, we isolated the gene, designated CET1, encoding the S. cerevisiae capping enzyme beta subunit. Amino acid sequence analysis revealed that the gene encodes for 549 amino acids with a calculated M(r) of 61,800 which is unexpectedly smaller than the size estimated by SDS-PAGE. Gene disruption experiment showed that CET1 is essential for yeast cell growth. The purified recombinant CET1 gene product, Cet1, exhibited an RNA 5'-triphosphatase activity which specifically removed the gamma-phosphate from the triphosphate terminated RNA substrate, but not from nucleoside triphosphates, confirming the identity of the gene. Interaction between the Cet1 and the Ceg1 was also studied by the West-Western procedure using recombinant Ceg1-[32P]GMP as probe. PMID- 9345281 TI - Induction of tissue-type plasminogen activator (tPA) and type-1 plasminogen activator inhibitor (PAI-1) as early growth responses in rat hepatocytes in primary culture. AB - Early growth response with respect to tissue-type plasminogen activator (tPA) and type-1 plasminogen activator inhibitor (PAI-1) gene expression was studied in rat hepatocytes in primary culture. The genes for tPA and PAI-1 could be categorized as a delayed early growth response (DER) gene and an immediate early growth response (IER) gene, respectively. The expression of tPA was much higher in growth-promoting than in static culture conditions (i.e., cultured at low density and/or on a collagen-coated dish), and that of PAI-1 was regulated in the opposite direction. Experiments using dibutyryl cAMP (dbcAMP) and H-89 showed that the cAMP/A-kinase system might be involved in the induction of the early growth response of tPA and in the augmentation of PAI-1 mRNA induction by dbcAMP. These fibrinolytic components, whose expression is closely associated with hepatocyte growth, may play important roles in pathophysiological events in the liver such as liver regeneration. PMID- 9345282 TI - Delayed Ca2+ response to glucose in diabetic GK rat. AB - The spontaneously diabetic non-obese GK (Goto-Kakizaki) rat exhibits high basal plasma glucose and insulin levels and poor glucose-induced insulin secretion, which makes it a suitable model for non-insulin dependent diabetes mellitus, NIDDM. The aim of this study was to investigate the handling of cytosolic free Ca2+ concentration ([Ca2+]i), the key regulator of insulin secretion, in GK rat single pancreatic islets. For this purpose the influence of high glucose (16.7 mM) and arginine (20 mM) on [Ca2+]i was studied in GK and Wistar rat islets, which served as controls. The data obtained suggest that glucose which through its metabolism generates ATP needed for closure of the KATP channels and membrane depolarization, induces a delayed [Ca2+]i response in the GK rat pancreatic islet. This delay in [Ca2+]i response is likely to result from a defective metabolism of glucose in the diabetic islet. PMID- 9345283 TI - Actinomycin D amplifies site-specific DNA cleavage induced by neocarzinostatin. AB - Neocarzinostatin (NCS) is an antineoplastic antibiotic causing DNA cleavage. Actinomycin D (ActD) is an antineoplastic antibiotic, which does not cleave DNA strands. We examined the mechanism of ActD-mediated amplification of NCS-induced DNA cleavage using 32P-labeled DNA fragments obtained from the human c-Ha-ras-1 proto-oncogene. NCS plus glutathione caused DNA cleavage at thymine and adenine residues in the absence of ActD. The addition of ActD enhanced the DNA cleavage at the sites at which the lesions on opposite strands are staggered 2 bases in a 3' direction, particularly at the 5'TCT-3'.3'-AGA-5', 5'-TGT-3'.3'-ACA-5' and 5' ACT-3'.3'-TGA-5' sequences, suggesting that ActD amplified double-stranded DNA cleavage. The mechanism of ActD-mediated amplification of NCS-induced DNA cleavage can be explained by assuming that binding of ActD to DNA changes the DNA conformation to allow NCS to bind to DNA at the specific sequences. PMID- 9345284 TI - mtDNA mutations confer cellular sensitivity to oxidant stress that is partially rescued by calcium depletion and cyclosporin A. AB - The complete mechanism by which pathogenic mtDNA mutations cause cellular pathophysiology and in some cases cell death is unclear. Oxidant stress is especially toxic to excitable nerve and muscle cells, cells that are often affected in mitochondrial disease. The sensitivity of cells bearing the LHON, MELAS, and MERRF mutations to oxidant stress was determined. All were significantly more sensitive to H2O2 exposure than their nonmutant cybrid controls, the order of sensitivity was MELAS > LHON > MERRF > controls. Depletion of Ca2+ from the medium protected all cell lines from oxidant stress, consistent with the hypothesis that death induced by oxidant stress is Ca(2+)-dependent. A potential downstream target of Ca2+ is the mitochondrial permeability transition, MPT, which is inhibited by cyclosporin A. Treatment of MELAS, LHON, and MERRF cells with cyclosporin A caused significant rescue from oxidant exposure, and in each case significantly greater rescue of mutant than control cells. The pronounced oxidant-sensitivity of mutant cells, and their protection by Ca2+ depletion and CsA, has potential implications for both the pathophysiological mechanism and therapy of these mitochondrial genetic diseases. PMID- 9345285 TI - The role of CD8+ Th2 lymphocytes in the development of smoking-related lung damage. AB - There is increasing evidence for the existence of different subsets of CD8+ T lymphocytes in humans, according to their cytokine secretion profile. Resistance or susceptibility to infections and the outcome of some inflammatory processes may depend on the lymphokine profile which predominates. We show one consequence of the switching of a host CD8+ T cell response from the Th1 effector function to the Th2 pattern in relation to the exposure to a common toxicant and its pathogenetic implications. Chronic obstructive bronchitis is a pulmonary disease characterized by airway inflammation with predominance of CD8+ T lymphocytes, mucus hypersecretion, repeated airway infections, and decline in lung function. Though smoking-related, it affects only a portion of smokers. The results of this study, comparing the functional characteristics of CD8+ T cell clones from smokers with the disease, unaffected smokers and healthy individuals, indicate that the smokers who have a predominance of CD8+ T lymphocytes of the Th2 phenotype may be predisposed to develop more severe smoking-induced lung damage, with chronic airway inflammation, repeated infections and persistent airflow obstruction. PMID- 9345286 TI - A stable partly denatured state of trypsin induced by high hydrostatic pressure. AB - The effect of hydrostatic pressure on the unfolding of trypsin was studied by fluorescence spectroscopy under pressure from 1 to 7000 bar. It was found that, at pH 3.0 or pH 7.3, a stable partly denatured state of trypsin was obtained when the applied pressure was about 6.5 kbar. This transient denatured state did not show any enzymatic activity and was different from that denatured by 8 M urea or high temperature in both intrinsic fluorescence spectrum and 8-anilino-1 naphtalene sulfonate (ANS) binding, having some obvious characteristics of 2 molten globule state of protein. It was also found that the formation of this partly denatured state of trypsin was temperature dependent. Energenic values of the process were also given. PMID- 9345287 TI - Effects of PTH on PTHrP gene expression in human osteoblasts: up-regulation with the kinetics of an immediate early gene. AB - Parathyroid hormone-related peptide (PTHrP) is a local regulator of human bone turnover, which shares some sequence homology with the systemic hormone parathyroid hormone (PTH). The two proteins exert cellular effects through interaction with a common G protein-coupled PTH/PTHrP receptor on target cells. Whilst the PTH gene has a relatively simple structure the PTHrP gene is complex, alternative splicing of which can generate multiple mRNA species encoding PTHrP of 139, 141 and 173 amino acids. To date little is known regarding the extent to which PTH and PTHrP interact to modulate bone cell function. In this study we have used the quantitative technique of Real-Time polymerase chain reaction (PCR) to investigate the ability of PTH to induce PTHrP expression in SaOS-2 cells and in primary human osteoblasts. In addition, we have used the semi-quantitative techniques of PCR followed by Southern analysis and scanning densitometry to investigate the effects of PTH(1-34) on expression of mRNA species encoding the three PTHrP isoforms. We report a 50 fold increase in PTHrP mRNA expression 30 min after treatment with 100 ng/ml human recombinant PTH (1-34) in SaOS-2 cells, and a 38 fold rise in human osteoblasts 45-90 min post-PTH treatment. mRNA species encoding for PTHrP 1-139, 1-141 and 1-173 were all induced in human osteoblasts 45 min after exposure to PTH. Whilst the 1-139 mRNA species exhibited a sustained expression, both the 1-141 and 1-173 isoforms showed a biphasic induction with a second peak 6 hr post PTH treatment. These data demonstrate that PTH induces expression of the PTHrP gene in both SaOS-2 and primary human osteoblasts with the kinetics of an immediate early gene. Up-regulation of the PTHrP gene in response to PTH may be an important physiological mechanism by which this systemic factor effects a localised response in bone. PMID- 9345288 TI - Bioconjugation of tumor necrosis factor-alpha with the copolymer of divinyl ether and maleic anhydride increasing its antitumor potency. AB - To enhance the therapeutic usefulness of antitumor cytokines in vivo, we synthesized bioconjugated tumor necrosis factor-alpha (TNF-alpha) with divinyl ether and maleic anhydride copolymer (DIVEMA), which has intrinsic antitumor activity as a synthetic biological response modifier. The degree of modification could be controlled by the addition of 2,3-dimethylmaleic anhydride (DMMAn), which binds to amino groups of TNF-alpha by changing the pH. In addition, the specific activity of DIVEMA-TNF-alpha was hardly decreased in vitro. DIVEMA-TNF alpha showed a marked antitumor effect compared to native TNF-alpha without any side effects such as sudden death, body-weight reduction, and decrease in platelet count on mice bearing solid tumors. These results suggest that DIVEMA is a useful polymeric modifier for-bioconjugation of TNF-alpha in order to increase its antitumor activity, and multifunctionally bioconjugated TNF-alpha may be a potentiated antitumor agent for therapeutic use. PMID- 9345289 TI - Extraction and stabilization of mammalian CDP-diacylglycerol synthase activity. AB - CDP-diacylglycerol synthase, also known as CTP: phosphatidic acid cytidylyltransferase (EC 2.7.7.41), is thought to be the rate-limiting enzyme in the synthesis of the inositol phospholipids, phosphatidylglycerol and cardiolipin. Its role in inositol phospholipid synthesis suggests its potential as a regulator of signal transduction as well. Although the mammalian cDNA for the synthase has recently been cloned, attempts to purify this enzyme from a mammalian source have been unsuccessful due to its lability in detergents. We report here the extraction and stabilization of CDP-diacylglycerol synthase from rat liver. Using a buffer containing 2M KCL, we were able to extract virtually all of the activity from microsomal membranes. This extract was stable indefinitely at -72 degrees C and for at least 24 hrs at 4 degrees C. Incubation at room temperature for 24 hours resulted in the loss of mor than half the activity. All detergents tested destroyed the activity. The activity was dependent on both substrates (phosphatidic acid and CTP) as well as on MgCl2, and inhibited by the product, CDP-diacylglycerol. Addition of GTP enhanced the activity approximately 2 fold, and bovine serum albumin increased activity by 6 fold. PMID- 9345290 TI - Role of protein kinase C-alpha in activation of ecto-5'-nucleotidase in the preconditioned canine myocardium. AB - We have reported that activation of protein kinase C (PKC) increases ecto-5' nucleotidase activity, which may contribute to the infarct size-limiting effect of ischemic preconditioning. Since we have reported that Ca(2+)- and phospholipid sensitive PKC is activated due to ischemic preconditioning, we further tested 1) whether PKC-alpha or -beta is translocated to the cellular membrane of the preconditioned canine myocardium, and 2) whether activation of PKC contributes to the increase in ecto-5'-nucleotidase activity via phosphorylation-dependent mechanisms. Four times of 5 minutes coronary occlusion separated by 5 minutes of reperfusion (ischemic preconditioning) translocated PKC-alpha to the cellular membrane in the canine hearts, although PKC-beta, -delta, -epsilon, and -zeta were not translocated. The activity of Ca(2+)- and phospholipid-sensitive PKC increased, which was attenuated by the removal of either Ca2+ or phosphatidylserine. Ecto-5'-nucleotidase was also activated in the preconditioned myocardium compared with control. Inhibition of PKC due to GF109203X blunted the activation of myocardial ecto-5'-nucleotidase. Okadaic acid (an inhibitor of phosphatase) enhanced the increases in ecto-5'-nucleotidase activity due to preconditioning, and this enhancement was blunted by GF109203X. We conclude that ischemic preconditioning activates PKC-alpha, and thus ecto-5'-nucleotidase. PMID- 9345291 TI - Molecular cloning and characterization of a novel TBP-1 interacting protein (TBPIP):enhancement of TBP-1 action on Tat by TBPIP. AB - The human immunodeficiency virus-1 (HIV-1) protein Tat, encoded by one of the HIV regulatory genes, tat, is reported to be essential for HIV gene expression and replication in infected cells. Observations suggest that several cellular factors cooperate with Tat in this process. Tat binding protein-1 (TBP-1) is reported to be one such cellular factor that specifically suppresses Tat-mediated transactivation of HIV replication in vitro. Here we have cloned a novel factor, TBP-1 interacting protein (TBPIP) from the mouse, which interacts with mouse TBP 1. TBPIP contains several kinase phosphorylation sites and co-localizes with TBP 1 in vivo. The fact that Tat activity is altered synergistically by the TBP-1 and an additional TBP-1 binding protein has not been reported before. We provide evidence that expression of TBPIP enhances the inhibitory action of TBP-1 on Tat mediated transactivation in vitro. Our results suggest that TBPIP may have a key role in suppressing the Tat-mediated transactivation. PMID- 9345292 TI - The tyrosine residue at 1250 of the insulin-like growth factor I receptor is required for ligand-mediated internalization. AB - The twin tyrosine residues at 1250 and 1251 of the insulin-like growth factor I receptor (IGF-IR) are missing in the corresponding homologous region of the insulin receptor. In this unique region, the tyrosine at 1251 (Y1251) is essential for both transforming and antiapoptotic activities of the IGF-IR, while Y1250 is dispensable for either of these functions. We show here that a receptor with a mutation at Y1250, but not at Y1251, has lost the ability for ligand mediated internalization when the mutant receptors are overexpressed in R- cells, derived from a mouse embryo with a targeted disruption of the IGF-IR gene. These results provide evidence that each twin tyrosine at Y1250 and Y1251 of the IGF-IR separately exerts different roles in biologically important signal transductions. PMID- 9345293 TI - An "in vitro" system simulates in membranes the antibacterial mechanism postulated for the action of isoxazolylnaphtoquinoneimine in Staphylococcus aureus. AB - The 2-hydroxy-N-(3,4-dimethyl-5-isoxazolyl)-1,4-naphthoquinone-4-imine (Q1) revealed good activity against Staphylococcus aureus. Q1 in contact with the bacteria experimented reduction evidenced by changes in its spectrum of absorption simultaneously with loss of colour. During the first 4 hours of incubation, oxygenation restored the original spectrum. Treatment with sodium borohydrure reduces irreversibly Q1. Redox-reaction "in vitro" was detected between Q1 and NADH in the presence of diaphorase. The environment of the probable site of action of Q1 was simulated using an artificial membrane system, instead of S. aureus membranes. Q1 interacts with lisophosphatidylcholine micelles following a cooperative binding model. The kinetics of Q1-reduction was increased by lipid micelles incorporated with the antibacterial compound. PMID- 9345294 TI - Solution structure by site directed tryptophan fluorescence in tear lipocalin. AB - The solution structure of the G strand of human tear lipocalin was deduced by site directed tryptophan fluorescence (SDTF). The fluorescent amino acid, tryptophan, was sequentially substituted for each native amino acid in the sequence of the G strand. The fluorescent properties resolved alternating periodicity as predicted for beta sheet structure, twists in the beta sheet, strand orientation in the lipocalin cavity, and the relative depth of residues in the cavity. A distribution of microstates with various orientations of dipoles in the side chain environments of the G strand revealed mobility on the nanosecond time scale. SDTF is broadly applicable to most proteins and will complement x-ray crystallography, site directed spin labeling by electron paramagnetic resonance (EPR), and nuclear magnetic resonance (NMR) in the determination of solution structure. PMID- 9345295 TI - Molecular characterization, expression in Escherichia coli, and epitope analysis of a two EF-hand calcium-binding birch pollen allergen, Bet v 4. AB - Birch pollen belongs to the most potent elicitors of Type I allergic reactions in early spring. Using serum IgE from a birch pollen allergic patient, two cDNA clones (clone 6 and clone 13) were isolated from a birch pollen expression cDNA library constructed in phage lambda gt11. Clone 6 encoded a 9.3 kD two EF-hand calcium-binding protein, designated Bet v 4, with significant end to end sequence homology to EF-hand calcium-binding allergens from weed and grass pollen. Recombinant Bet v 4, expressed as beta-galactosidase fusion protein, reacted with serum IgE from approximately 20% of pollen allergic individuals. Depletion of allergenbound calcium by EGTA treatment lead to a substantial reduction of IgE binding to Bet v 4, indicating that protein-bound calcium is necessary for the maintenance of IgE-epitopes. The greatly reduced IgE-binding capacity of clone 13, a Bet v 4 fragment that lacked the 16 N-terminal amino acids, indicated that the N-terminus contributes significantly to the proteins IgE-binding capacity. By IgE-inhibition experiments it was demonstrated that recombinant Bet v 4 shared IgE-epitopes with natural Bet v 4 and a homologous timothy grass pollen allergen. Recombinant Bet v 4 may therefore be considered as a relevant crossreactive plant allergen, which may be used for diagnosis and treatment of patients suffering from multivalent plant allergies. PMID- 9345297 TI - Regulation of superoxide anion radical-superoxide dismutase system in the avian thyroid by TSH with reference to thyroid hormonogenesis. AB - This study shows that superoxide dismutase is present in the thyroid gland of pigeons as a constitutive enzyme serving as an antioxidant against oxygen toxicity. Exogenous administration of thyrotropin induced thyroidal superoxide dismutase with a simultaneous burst in superoxide anion radical levels during the initial phase of hormone treatment. The superoxide radical generated was completely scavenged by SOD during the late phase of TSH-treatment, presumably as an adaptive measure to check the oxygen burst. TSH failed to augment serum T3 levels, although the thyroxine level in the serum was elevated. The peak level of SOD activity profile in the thyroid gland correlated very well with the peak level of thyroxine concentrations in the serum of pigeon. It is reasonable to postulate that the thyroidal SOD in homeotherms serves a dual role, firstly as a strategic antioxidant enzyme to protect the thyroid gland against the degenerative influence of toxic oxyradicals and secondly to provide H2O2 for thyroid hormone biosynthesis. Our results confirm the previous observations that TSH is mainly thyrotropic in birds and that it has no influence on the peripheral activation of thyroxine to triiodothyronine by stimulating the extra thyroidal 5' deiodinase activity. PMID- 9345298 TI - Functional elements in molecular chaperone alpha-crystallin: identification of binding sites in alpha B-crystallin. AB - alpha-Crystallin, the predominant eye lens protein with sequence homology to small heat shock proteins, acts like a molecular chaperone by suppressing the aggregation of damaged crystallins and proteins. To gain an insight into the amino acid sequences in alpha-crystallin involved in chaperone-like function, we used a cleavable, fluorescent, photoactive, crosslinking agent, sulfosuccinimidyl 2 (7-azido-4-methylcoumarin-3-acetamido)-ethyl-1,3' dithiopropionate (SAED), to derivatize yeast alcohol dehydrogenase (ADH) and allowed it to complex with bovine alpha-crystallin at 48 degrees C. The complex was photolyzed and reduced with DTT and the subunits of alpha-crystallin, alpha A- and alpha B-, were separated. Fluorescence analysis showed that both alpha A- and alpha B crystallins interacted with ADH during chaperone-like function. Tryptic digestion, amino acid sequencing, and mass spectral analysis of alpha B crystallin revealed that APSWIDTGLSEMR (57-69) and VLGDVIEVHGKHEER (93-107) sequences were involved in binding with ADH. PMID- 9345296 TI - A synthetic peptide, FN-C/H-V, from the C-terminal heparin-binding domain of fibronectin promotes adhesion of PMA stimulated U937 cells. AB - Hematopoietic cells differentially bind to the C-terminal heparin-binding domain of fibronectin depending on the activation state of integrin alpha 4 beta 1. In this study, we have identified a synthetic peptide derived from the C-terminal heparin-binding domain of fibronectin that promotes adhesion of PMA-treated U937 cells (a monocytic cell line) in a dose-dependent manner. A peptide (FN-C/H-V; residues Gln1892 to Gly1910) was active to inhibit adhesion of PMA-treated U937 cells to the 29-kDa fragment comprising the C-terminal heparin-binding domain of fibronectin. A peptide with scrambled version of FN-C/H-V lost the inhibitory activity on the adhesion. Furthermore, the IgG-conjugated FN-C/H-V promoted the adhesion of PMA-treated U937 cells to an extent comparable to that of the 29-kDa fragment. The adhesion of PMA-treated U937 cells on IgG-conjugated FN-C/H-V was inhibited both by anti-alpha 4 beta 1 antibody and by glycosaminoglycans including chondroitin sulfate and heparan sulfate. The other peptide, FN-C/H-II, was also a weak adhesion-promoting domain. These results suggest that the amino acid sequence defined by peptide FN-C/H-V contributes to the main adhesion promoting activity of the 29-kDa fragment of fibronectin to stimulated U937 cells. The regulation of interactions of alpha 4 beta 1 integrin and glycosaminoglycans with ligands in fibronectin may have important implications for the migration and function of U937 cells. PMID- 9345299 TI - Catalysis of guanine nucleotide exchange on eIF-2 by eIF-2B: is it a sequential or substituted enzyme mechanism? AB - The mechanism of action of the eukaryotic initiation factor eIF-2B in catalyzing the exchange of guanine nucleotides bound to eIF-2 is uncertain--evidence having been adduced for a sequential mechanism and for a substituted enzyme mechanism. Data purporting to support a substituted enzyme mechanism have been analysed and shown to be ambiguous and equally consistent with a sequential mechanism. Suitable rate constants for a sequential mechanism involving the transient formation of the quaternary complex eIF-2.eIF-2B.GDP.GTP are suggested. PMID- 9345300 TI - The pkI gene encoding pyruvate kinase I links to the luxZ gene which enhances bioluminescence of the lux operon from Photobacterium leiognathi. AB - Partial 3'-end nucleotide sequence of the pkI gene (GenBank accession No. AF019143) from Photobacterium leiognathi ATCC 25521 has been determined, and the encoded pyruvate kinase I is deduced. Pyruvate kinase I is the key enzyme of glycolysis, which converts phosphoenol pyruvate to pyruvate. Alignment and comparison of pyruvate kinase Is from P. leiognathi, E. coli and Salmonella typhimurium show that they are homologous. Nucleotide sequence reveals that the pkI gene is linked to the luxZ gene that enhances bioluminescence of the lux operon from P. leiognathi. The gene order of the pkI and luxZ genes is-pk1-ter--> R&R"-luxZ-ter"-->, whereas ter is transcriptional terminator for the pkI and related genes, and R&R" is the regulatory region and ter" is transcriptional terminator for the luxZ gene. It clearly elicits that the pkI gene and luxZ gene are divided to two operons. Functional analysis confirms that the potential hairpin loop omega T is the transcriptional terminator for the pkI and related genes. It infers that the pkI and related genes are simply linked to the luxZ gene in P. leiognathi genome. PMID- 9345301 TI - Impact of mutations at different serine residues on the tyrosine kinase activity of the insulin receptor. AB - Insulin binding to its receptor activates a cascade of signaling events which are initiated by tyrosine autophosphorylation of the receptor and activation of the tyrosine kinase activity towards the insulin receptor substrates. In addition to phosphorylation at tyrosine residues a serine phosphorylation of the insulin receptor is observed. Neither the functional significance of serine phosphorylation of the receptor nor the location of relevant regulatory sites has been determined exactly so far. We studied potential functions of serine residues in human insulin receptor (HIR) with respect to its ability to undergo insulin stimulated autophosphorylation. Using site directed mutagenesis of HIR we exchanged serine to alanine at 13 different positions in the HIR beta-subunit. Sites were chosen according to the criteria of known serine phosphorylation sites (1023/25, 1293/94, 1308/09), conserved positions in hIR, hIGF-1 receptor, hIRR, and dIR (962, 994, 1037, 1055, 1074/78, 1168, 1177/78/82, 1202, 1263, 1267). All HIR mutants were expressed in HEK 293 cells and basal and insulin stimulated autophosphorylation were determined. We found that the exchange of serine to alanine at position 994 and at position 1023/25 increased insulin stimulated receptor autophosphorylation significantly (147% +/- 12% and 129% +/- 6% of control, p < 0.01, n = 7), while all other exchanges did not significantly alter insulin stimulated HIR autophosphorylation. The data suggest that the serine residues at position 994 as well as 1023/25 might be part of inhibitory domains of the insulin receptor. PMID- 9345302 TI - Synthetic genes specifying periodic polymers modelled on the repetitive domain of wheat gliadins: conception and expression. AB - In order to optimise new polypeptide based biomaterials, we developed a procedure for producing homoblock polypeptides using recombinant DNA technology. Synthetic genes encoding periodic polypeptides modelled on the consensus sequence of wheat gliadins (a family of wheat storage proteins) were devised to be expressed in Escherichia coli. The construction strategy followed allows the construction of three genes encoding 8, 16, and 32 copies of the PQQPY module. The optimal expression conditions in the enterobacteria were established and a convenient purification procedure was shown to be useful in recovery of sizable amounts of strictly periodic polypeptides. The identities of the synthesized polypeptides were assessed using positive cross reactions to antibodies raised against a synthetic decapeptide (PQQPYPQQPA) and amino acid composition was determined as well. PMID- 9345303 TI - Trafficking kinetics of the insulin-regulated membrane aminopeptidase in 3T3-L1 adipocytes. AB - In fat and muscle cells insulin causes the marked translocation of the glucose transporter GLUT4 from its intracellular location to the plasma membrane. We and others have discovered an insulin-regulated membrane aminopeptidase (designated IRAP) that colocalizes with intracellular GLUT4 and also translocates markedly in response to insulin. This study describes the trafficking kinetics of IRAP in 3T3 L1 adipocytes. By means of a surface biotinylation method, the half-time for the increase in IRAP at the plasma membrane in response to insulin was found to be 2 min. The increase was completely blocked by the phosphatidylinositol 3-kinase inhibitor, wortmannin. In insulin-treated cells, biotinylated IRAP, initially at the plasma membrane, equilibrated with the intracellular pool with a half-time of 2 min. Thus, IRAP continuously recycles. Finally, vesicles isolated from the intracellular membranes with antibodies against IRAP and GLUT4 showed the same protein composition. In conjunction with results in the literature, these findings indicate that IRAP and GLUT4 traffic through the same intracellular compartments. PMID- 9345304 TI - A conditional version of the Ets transcription factor Erm by fusion to the ligand binding domain of the oestrogen receptor. AB - The fusion of a wide range of proteins to the ligand-binding domain of nuclear receptors has been shown to impart ligand-dependent inducible activity of the resulting chimera. Transcriptional regulators of the ETS family are involved in both normal and oncogenic processes. In order to address the role of Erm, a "PEA3 subgroup" member of this family, we generated a chimera between Erm and the widely used ligand-binding domain of the oestrogen receptor (ER). The chimera, ErmER, consists of Erm protein fused at its C-terminal end to the ER domain. We show that ErmER displays a ligand-dependent transcriptional activity on ets responsive elements. The efficiency of ErmER mediated transactivation is modulated by the hormone concentration while its weak leakiness is reduced by using the steroidal anti-oestrogen EM-139. Our results define ErmER as the first conditional version of an Ets transcription factor, providing a useful tool to decipher Erm biological role and to identify potential Erm target genes. PMID- 9345305 TI - Isolation and characterization of mitochondrial DNA-less lines from various mammalian cell lines by application of an anticancer drug, ditercalinium. AB - Since ethidium bromide was not effective in mouse cell lines for isolating mitochondrial DNA (mtDNA)-less cells (rho zero cells), we examined whether an anticancer drug, ditercalinium (DC), which has been shown to exclude mtDNA from mouse cell lines, could be effective in various mouse and human cell lines. We found that after DC treatment rho zero cells could be isolated from all cell lines of mouse or human origin tested. Moreover, these rho zero cells maintained ability to receive exogenously imported mtDNA and allow its replication and gene expression. These observations suggest that DC eliminates mtDNA from mouse and human cells without affecting the property to receive exogenous mtDNA. Therefore, DC could be applicable to cell lines expressing various differentiated phenotypes for studying whether mtDNA plays a significant role in expression of phenotypes by manipulating mtDNA elimination and reintroduction. PMID- 9345306 TI - cDNA cloning and mRNA expression of the human adrenoleukodystrophy related protein (ALDRP), a peroxisomal ABC transporter. AB - We have cloned the cDNA containing the complete coding region of the human adrenoleukodystrophy related (ALDR) gene. The 2220-bp open reading frame encodes a 740-amino-acid polypeptide with a predicted molecular weight of 83.3 kDa. The human ALDR protein displays high similarity (62.8% identical amino acid residues) to the human adrenoleukodystrophy (ALD) gene. Analysis of ALDR expression revealed the presence of ALDR mRNA in a variety of human tissues, predominantly in brain and heart. This expression pattern is different from all other known peroxisomal ABC-transporters. Defects in the ALD gene are the primary cause of adrenoleukodystrophy, a demyelinating disorder of the central nervous system. The ALD protein (ALDP) and the ALDR gene product are peroxisomal membrane proteins belonging to the superfamily of transporters containing an ATP-binding cassette (ABC-transporters). All known peroxisomal ABC-transporters represent only one half of a functional transporter. They are expected to form dimers either as a homodimer or as a heterodimer. ALDRP is a potential dimerization partner of ALDP or other peroxisomal ABC-transporters. The ALDR gene is a candidate for a modifier gene, accounting for the strikingly varying clinical courses of ALD observed even within a family. PMID- 9345307 TI - Renal elimination of islet amyloid polypeptide. AB - Six healthy volunteers showed significantly higher plasma islet amyloid polypeptide levels following an oral glucose tolerance test compared to fasting levels. The urine IAPP concentration before and after the OGTT was comparable to that in plasma. Reverse phase HPLC and radioimmunoassay analysis of urine samples revealed a single IAPP-immunoreactive peak. Before hemodialysis, the plasma levels of IAPP and C-peptide, but not of insulin, were significantly elevated in eight fasting patients with chronic renal failure, compared to eight healthy matched control subjects. After hemodialysis, there was a tendency for decreased IAPP levels compared to before dialysis. In summary, elevated levels of plasma IAPP were found in patients with chronic renal failure and the peptide is eliminated by hemodialysis. Furthermore, immunoreactive IAPP is normally present in the urine. These results suggest that IAPP is, at least in part, renally eliminated from the plasma by excretion (glomerular filtration and/or tubular secretion. PMID- 9345308 TI - Broad ligand specificity of the transcriptional regulator of the Bacillus subtilis multidrug transporter Bmr. AB - The expression of the Bacillus subtilus multidrug-efflux transporter Bmr can be induced by two of its structurally dissimilar substrates, rhodamine 6G and tetraphenylphosphonium, through their direct interaction with the transcriptional regulator BmrR (Ahmed et al., J. Biol. Chem. 269, 28506). Here, by screening a chemical library, we identified four additional ligands of BmrR inducing Bmr expression at micromolar concentrations. BmrR ligands, although sharing a positive charge and moderate hydrophobicity, are structurally very diverse. At the same time, not all hydrophobic positively charged compounds, including many structural analogs of the inducers, induce Bmr expression, thus suggesting that local chemical interactions and not merely physical properties of the ligands are important for their recognition by BmrR. These results confirm that this soluble protein, like the membrane transporter it regulates, has a uniquely broad substrate specificity. PMID- 9345309 TI - Differential effects of interleukin-1 beta, interleukin-2, and interferon-gamma on the inducible expression of CYP 1A1 and CYP 1A2 in cultured rabbit hepatocytes. AB - The effects of interleukin-1 beta, interleukin-2 and interferon-gamma and their combinations were investigated on induced cytochrome P 4501A of cultured rabbit hepatocytes considered 72 h after plating. Without apparent cellular toxicity, these cytokines provoke a significant decrease in TBZ- and BNF-induced P4501A1/ 2 expression. However specific patterns of action are revealed: IL-1 beta is the most potent cytokine in regard to CYP1A1/2 mRNA suppression whereas IL-2 exerts repressive effects only on P4501A1 induced expression. Although being a strong inhibitor of induced enzymatic activities and protein contents, IFN-gamma exhibits only a weak influence on CYP1A1/2 mRNAs with the exception of its association with interleukins. All these results suggest that IL-1 beta and IL-2 promote mainly transcriptional repression mechanism whereas IFN-gamma would stimulate a post-transcriptional suppressive pathway. PMID- 9345310 TI - Differential expression of mammalian TRP homologues across tissues and cell lines. AB - Mammalian homologues of the Drosophila trp gene have been invoked as the structural basis for the currents associated with capacitative Ca2+ entry (CCE) in many cell types. Trp homologues are members of a large protein family that may associate as channel subunits providing an explanation for the functional diversity of store-operated channels observed in these cells. However, there is little information as to which of these genes are co-expressed at the cellular level. We have examined the tissue specific expression of five mammalian trp genes and determined which are co-expressed in five different cell lines. The results show tissue- and cell-specific co-expression of multiple trp forms. This implies that the subunit composition of a particular CCE channel may vary depending on the cell type. PMID- 9345311 TI - Dissociation of nitric oxide from soluble guanylate cyclase. AB - Kinetic studies of soluble guanylate cyclase complexed with nitric oxide prove that NO dissociation in the presence of the substrate GTP and Mg2+ is as much as 50 times faster than in their absence. In the presence of those two reagents the dissociation rate constant is k(obs) = 0.04 +/- 0.01 s-1 at 20 degrees C, which is by far the fastest NO dissociation rate constant ever reported for a ferrous heme protein. Extrapolated to 37 degrees C, this corresponds to a half life of about 5 s for NO dissociation from soluble guanylate cyclase at physiological conditions, which is presumably fast enough to account for deactivation of the enzyme in biological systems. Dissociation rate constants are also reported for a variety of other reagent conditions. PMID- 9345312 TI - Neuropeptide Y enhances ATP-induced formation of inositol phosphates in chromaffin cells. AB - Bovine chromaffin cells contain high affinity NPY binding sites coupled through a pertussis toxin-sensitive G protein to inhibition of cAMP accumulation. NPY alone does not alter [3H]inositol phosphate formation from [3H]phosphoinositides in these cells. Increasing NPY concentrations, in the presence of ATP (300 microM), produced a dose-dependent enhancement in [3H]-inositol phosphate formation, EC50 = 3.2 nM. Inclusion of the selective NPY-Y1 receptor antagonist BW1229 (1 microM) produced a marked decrease in NPY potency (EC50 = 3.3 microM). The Y1 receptor agonist, [Leu31, Pro34]-NPY, was equally effective with NPY, whereas NPY18-36, a Y2 receptor agonist, was much less effective. Inclusion of NPY with ATP also produced an enhancement in the release of intracellular Ca2+. The ability of NPY to enhance both [3H]inositol phosphate formation and the release of intracellular Ca2+ was pertussis toxin-insensitive. NPY action on bovine chromaffin cell receptor(s) appears to facilitated by different G proteins: one which can inhibit cAMP accumulation via a pertussis toxin-sensitive process and another which can enhance ATP activation of the inositol phosphate signaling pathway by a pertussis toxin-insensitive process. PMID- 9345313 TI - Chaperonin genes of the Synechocystis PCC 6803 are differentially regulated under light-dark transition during heat stress. AB - Transcriptional startpoints of the two heat inducible chaperonin genes of Synechocystis PCC 6803 were mapped within the conservative CIRCE element and proved to be identical irrespective of the temperature treatment. Finding of an ORF encoding for a potential CIRCE binding repressor (HrcA) further suggests that both groEL-analogs are regulated in a CIRCE-dependent manner. In contrast to the expectations, the chaperonin twins are differentially expressed under light-dark transition during heat stress. Not the light per se, but rather the photosynthetic electron transport appears to be accountable for the regulatory differences. Our findings support the hypothesis that multiple chaperonins play different physiological roles under stress conditions. PMID- 9345314 TI - Phenol sulfotransferase pharmacogenetics in humans: association of common SULT1A1 alleles with TS PST phenotype. AB - The phenol sulfotransferases (PSTs) catalyze the sulfation of both small planar phenols and phenolic monoamines. Three highly homologous PST genes, SULT1A1, SULT1A2, and SULT1A3, are known to exist in humans. The prototypic biochemical phenotype associated with the enzyme encoded by SULT1A1 is the thermal stable (TS) sulfation of 4 microM 4-nitrophenol (TS PST activity). Biochemical pharmacogenetic studies have demonstrated that individual variation in both TS PST activity and thermal stability in humans are inherited. As a step toward understanding molecular mechanisms responsible for the genetic regulation of PSTs in humans, we report here common SULT1A1 nucleotide polymorphisms that are associated with phenotypic variation in both platelet TS PST activity and thermal stability. When 905 human subjects were phenotyped for platelet TS PST activity and thermal stability, activity varied more than 50-fold, and thermal stability varied over 10-fold. DNA was isolated from the blood of 33 of these subjects selected on the basis of "extreme" TS PST phenotypes: high activity and high thermal stability; low activity and low thermal stability; or low activity and high thermal stability. These 33 subjects were genotyped for SULT1A1 by PCR amplification and sequencing of the entire open reading frame (ORF) as well as approximately 1 kb of intron DNA sequence. One common allele, SULT1A1*2, was uniformly associated with both very low TS PST activity and low thermal stability. The allele frequency of SULT1A1*2 in a randomly selected population sample of 150 Caucasian blood donors was 0.31 (31%), indicating that approximately 9% of this population would be homozygous for that allele. PMID- 9345315 TI - RpoS dependent overexpression of carotenoids from Erwinia herbicola in OXYR deficient Escherichia coli. AB - Carotenoid synthesis in Escherichia coli, when transformed with plasmid containing a carotenoid gene cluster from Erwinia herbicola (pPL376), is regulated by RpoS. When the plasmid was transformed into E. coli mutants that were oxyR minus, the intracellular carotenoid concentration dramatically increased from that observed in an oxyR plus allele. The higher carotenoid concentration in these mutants correlated with an increase in rpoS transcription as indicated by beta-galactosidase activity from a rpoS::lacZ promoter fusion. This indication of a higher concentration of carotenoids correlated with an increased resistance to hydrogen peroxide and near-ultraviolet radiation (310-400 nm; near-UV). PMID- 9345316 TI - Inhibition of inducible nitric oxide synthase expression and stimulation of the endothelial formation of nitric oxide most likely accounts for the protective effect of 2-(allylthio)pyrazine in a murine model of endotoxemia. AB - The lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS) in the vascular wall accounts, at least in part, for the severe hypotension in endotoxemia. The present study investigated whether 2 (allylthio)pyrazine (2-AP), an antioxidant, affects the LPS-induced expression of iNOS in rat aortic rings and the LPS-induced mortality in mice. 2-AP prevented the LPS-induced attenuation of contractions to phenylephrine, formation of cyclic GMP, and expression of iNOS in aortic rings without endothelium and caused endothelium-dependent nitric oxide-mediated relaxations. The mortality of mice receiving a lethal bolus of LPS was decreased by 2-AP, and this effect was associated with a reduced serum nitrite and nitrate level. These findings suggest that agents which inhibit the expression of iNOS but stimulate the formation of endothelium-derived nitric oxide may be of therapeutical value for the treatment of endotoxemia. PMID- 9345317 TI - Phenylarsine oxide inhibits insulin activation of phosphatidylinositol 3'-kinase. AB - Two early events downstream of insulin receptor autophosphorylation that are necessary for activation of glucose transport in adipocytes appear to be: (1) The tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) which (2) recruits and activates phosphatidylinositol 3'-kinase (PI3'-K). Phenylarsine oxide (PAO) has long been known to inhibit glucose transport, without inhibiting insulin receptor auto- or substrate phosphorylation. However, the PAO-sensitive site downstream of these early regulatory eventshas not been identified. Here we provide evidence that exposure of 3T3-L1 adipocytes to PAO inhibits PI3'-K activation, but it does not decrease either IRS-1 tyrosine-phosphorylation or the recruitment of PI3'-K to IRS-1 after insulin stimulation. PAO is also shown to inhibit PI3'-K activity in vitro. Therefore, since PI3'-K activation is essential for insulin stimulation of glucose transport, our results demonstrate that PI3'-K is a PAO-sensitive target of the insulin signaling pathway regulating glucose transport. PMID- 9345318 TI - Spatial orientation of tissue-type plasminogen activator bound at the melanoma cell surface. AB - Human melanoma cells produce tissue-type plasminogen activator (tPA) which is bound to the cell surface where it effectively mediates generation of plasmin. The present study is focused on analysis of involvement of the tPA domains in binding of the enzyme to the cell surface. The extent of plasminogen activation by tPA of melanoma cells was measured using an immunocapture assay. The activator anchored to solid surface via monoclonal antibodies directed to the individual domains of the activator exhibited variable enzymatic activity. The tPA was the most effective when bound by the antibodies against kringle-1 or kringle-2. Accessibility of the epitopes within cell surface-bound tPA was probed by the same set of monoclonal antibodies. FACS analysis showed that the epitopes within the finger/growth factor domain one part of the kringle-2 domain and the active site epitope were the most exposed. The kringle-1 domain epitope and the protease region epitope appeared partially exposed. Full-length melanoma-derived tPA and three recombinant domain-deletion variants of tPA were compared for their capacity to bind to the melanoma cells. The estimated IC50 value for the melanoma derived tPA was 2.3 +/- 0.25 microM. Comparable IC50 values were found for the tPA variant lacking the finger domain (3.6 +/- 0.6 microM) as well as for the variants consisting only of the kringle-2 and protease domains (7.5 +/- 0.45 microM). In contrast the value found for a tPA variant lacking the kringle-2 domain was > 100 microM. The consistent results obtained by the three different experimental approaches provide evidence that tPA binds to melanoma cells via its kringle-2 domain but binding sites within kringle-1 domain and protease domain may support the interaction. The finger domain did not contribute to the binding. PMID- 9345319 TI - Chiral discrimination of 2'-deoxy-L-cytidine and L-nucleotides by mouse deoxycytidine kinase: low stereospecificities for substrates and effectors. AB - The effects of four kinds of 2'-deoxy-L-nucleoside 5'-triphosphates and L-ATP, which are enantiomers of natural D-dNTPs and D-ATP, on deoxycytidine kinase (dCK) partially purified from mouse leukemic P388 cells were investigated. Only L-dCTP did not act as a phosphate donor while other L-dNTPs and L-ATP showed 15-30% of the activity of the corresponding D-dNTP or D-ATP. L-dCTP inhibited dCK non competitively with 2'-deoxycytidine (D-dCyd) and competitively with phosphate donor D-ATP. These inhibitory effects of L-dCTP on dCK were similar to the results of earlier studies using D-dCTP. Thus, L-dCTP was shown to be capable of serving as a feedback inhibitor for dCK instead of D-dCTP. Mouse dCK was also able to phosphorylate L-dCyd, as demonstrated in the case of human dCK. The present results suggest that the chirality of not only dCyd as the substrate but also nucleotides as the substrate or effector is not strictly discriminated by dCK. PMID- 9345320 TI - Novel cancer chemopreventive effects of a flavonoid antioxidant silymarin: inhibition of mRNA expression of an endogenous tumor promoter TNF alpha. AB - In this study we describe exceptionally high protective effects of silymarin, a flavonoid antioxidant isolated from milk thistle, against 12-O tetradecanoylphorbol 13-acetate (TPA)- and okadaic acid (OA)-caused tumor promotion in SENCAR mouse skin. Pre-application of silymarin to that of TPA in 7, 12-dimethylbenz(a)anthracene (DMBA)-initiated mouse skin resulted in almost complete protection in terms of tumor incidence (85%) as well as multiplicity (94%). In OA-caused tumor promotion studies, application of silymarin prior to that of OA in DMBA-initiated mouse skin resulted in a complete protection against tumorigenicity. We next assessed the effect of silymarin on TPA- and OA-caused induction of mRNA expression of tumor necrosis factor alpha (TNF alpha) which is an endogenous tumor promoter and a central mediator of tumor promotion in vivo in the case of both TPA and OA tumor promotion. Topical application of silymarin on mouse skin prior to that of TPA or OA resulted in a highly significant to complete inhibition in a dose-dependent manner against both TPA- and OA-caused induction of TNF alpha mRNA expression in mouse epidermis. These results indicate that silymarin exerts novel chemopreventive effects against tumorigenicity by inhibiting endogenous tumor promoter TNF alpha. Additional studies are warranted in other tumor models to further evaluate the cancer chemopreventive effect of silymarin and to define the involvement of TNF alpha as a molecular target for such an effect. PMID- 9345321 TI - Serum leptin levels are associated with hyperinsulinemia independent of body mass index but not with visceral obesity. AB - To examine the relationship between leptin levels and visceral obesity or plasma insulin levels, we studied serum leptin levels, fat distribution assessed by CT scan, and plasma insulin levels during 75 g oral glucose load in 100 Japanese men. Regression analysis adjusted by age and body mass index (BMI) showed leptin levels to be associated with visceral fat area(V)(p = 0.003), subcutaneous fat area(S)(p < 0.0001), and V + S(p < 0.0001), but not with V/S ratio(p = 0.897). By regression analysis adjusted by age, BMI, and V + S, serum leptin levels were still highly and positively correlated with plasma insulin levels during 75 g oral glucose load (p < 0.001), insulin resistance index(p < 0.001), and beta cell function index(p = 0.009) in homeostasis model assessment. These data suggest that hyperinsulinemia, but not visceral obesity, may be regulators of serum leptin levels independent of BMI. PMID- 9345322 TI - Inhibition growth of multidrug resistant KBV200 cells by MDR1 antisense RNA. AB - Acquisition of resistance to multiple drugs of tumor cell caused by overexpression of the MDR1 gene is one of major obstacles in cancer chemotherapy. We have attempted to reverse the multidrug resistance (MDR) phenotype by treating vincristine (VCR) and adriamycin (ADM) resistant KBV200 cells with MDR1 antisense RNA. Retroviral vector expressing the antisense RNA was transfected into KBV200. In the transfected cells, a stable expression of antisense RNA and a reduction of cellular MDR1 mRNA could be detected by RT-PCR, and a reduction of MDR1 specific P-glycoprotein (P-gp) was also detected by Western blot, whereas an increase of the drug concentration in the cells was detected by FACS. The IC50 of transfected cells to VCR and ADM was reduced by 65 and 47%. This study demonstrates that antisense RNA can increase the sensitivity of tumor cells to anticancer drug by decreasing the expression of the MDR1 gene. This strategy may be applicable to cure cancer patients with P-gp mediated MDR phenotype. PMID- 9345323 TI - Inositol-1,3,4,5-tetrakisphosphate binding sites in control and ras-transformed NIH/3T3 fibroblasts. AB - Inositol-1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4) binding properties were investigated in NIH/3T3 fibroblasts and its ras-transformant (DT cells), in which inositol tetrakisphosphates induce Ca2+ influx. [3H]-Ins(1,3,4,5)P4 bound to membranes of both types of cells with Kd values of 10.6 and 8.6 nM, respectively. The rank order of inositol polyphosphates for displacing [3H]Ins(1,3,4,5)P4 in DT cells was Ins(1,3,4,5)P4 > inositol-1,3,4,5,6-pentakisphosphate > inositol hexakisphosphate > inositol-1,4,5-trisphosphate. This order is similar to that reported in two Ras-GTPase-activating proteins, GAP1IP4BP and GAP1m, which are also the Ins(1,3,4,5)P4 binding proteins. Northern blot analysis revealed that NIH/3T3 and DT cells expressed mRNA species that were hybridizable with GAP1m cDNA. These results suggest that parental and ras-transformed NIH/3T3 fibroblasts possess GAP1-like proteins, which may be responsible for triggering inositol tetrakisphosphate-dependent Ca2 influx. PMID- 9345324 TI - Retinal 5-methoxytryptamine and 5-methoxyindole-3-acetic acid in the rat and quail: diurnal rhythms and interspecies differences. AB - Endogenous 5-methoxytryptamine (5MT) and its biosynthetic oxidation product, 5 methoxyindole-3-acetic acid (5MIAA), were successfully identified and measured in the retina of the rat and quail by gas chromatography/electron-capture negative ion chemical ionization mass spectrometry (GC/EC-NICI-MS). In the rat retina, diurnal rhythms of 5MT and 5MIAA, with high levels at mid-light and opposite to that of melatonin, were observed. In the quail, high levels of retinal 5MT and 5MIAA were found at mid-dark, and in phase to that of melatonin. Biosynthetic pathways for retinal 5MT and 5MIAA in the rat and quail were discussed in relation to the diurnal rhythms observed. Our results indicate that the biosynthesis and physiological functions of retinal 5MT and 5MIAA could be species dependent. PMID- 9345325 TI - Impact of adding concurrent chemotherapy to hyperfractionated radiotherapy for locally advanced non-small cell lung cancer (NSCLC): comparison of RTOG 83-11 and RTOG 91-06. AB - A hyperfractionated radiation therapy (HFX RT) trial (1.2 Gy twice daily, b.i.d.) (HFX) for non-small cell lung cancer (NSCLC) showed that 69.6 Gy resulted in better survival than did lower total doses (Radiation Therapy Oncology Group, RTOG 83-11) and that cisplatin concurrent with irradiation improved local control and survival over RT alone (Radiation Therapy Oncology Group, RTOG 91-06). Concurrent combination chemotherapy and HFX could improve both local and systemic control. In a phase II trial (RTOG 91-06) for inoperable NSCLC, two cycles of PE were used [cisplatin 50 mg/m2 intravenously (i.v.) days 1 and 8, etoposide 50 mg orally (p.o.) b.i.d., 75 mg/day if body surface area (BSA) < 1.7 m2, days 1-14] starting on day 1 of HFX (69.6 Gy) and repeated on day 29. HFX/PE was compared with HFX (69.6 Gy) from an earlier phase II trial (RTOG 83-11). Seventy-six patients treated with HFX/PE and 203 patients who received HFX alone were compared for toxicity, response, survival, and patterns of failure. The rates of grade 4 nonhematologic toxicity were similar (3.0% for HFX/PE, 3.0% for HFX), but grade 4 hematologic toxicity occurred only with HFX/PE 56.6%. Three (3.9%) HFX/PE patients had fatal toxicity (2 pulmonary, 1 renal); 1 HFX patient had fatal esophageal toxicity. Response and metastasis rates were similar for the two treatments, but infield (p = 0.054) and overall (p = 0.04) progression-free survival rates were better with HFX/PE. Median survivals were 18.9 months with HFX/PE and 10.6 months with HFX. Two-year survival rates were 36% for HFX/PE and 22% for HFX (p = 0.014). The differences in survival between HFX/PE and HFX remained borderline statistically significant (p = 0.0593) in the multivariate model, which included weight loss, Karnofsky performance status (KPS), sex, and stage. HFX/PE is an effective regimen in patients with inoperable NSCLC, although it is considerably more toxic, and is undergoing a comparison in a three-arm randomized phase III study against induction cisplatin/vinblastine plus standard once-daily RT and against cisplatin/vinblastine concurrent with standard RT. PMID- 9345326 TI - Postoperative radiation therapy in non-small cell lung cancer. AB - One hundred seventy-three patients with the diagnosis of non-small cell lung cancer (NSCLC) were treated with surgery and postoperative radiation therapy (RT) at the Radiation Oncology Center, Washington University, St. Louis. All patients had been retrospectively reviewed and restaged according to the new American Joint Committee (AJC) staging classification. Seventeen patients were stage I, 69 were stage II, 74 were stage IIIA, and 2 patients were stage IIIB. In 11 patients, accurate staging could not be determined. Indications for postoperative RT were positive surgical margins (32 patients), metastatic hilar nodes (78 patients), and metastatic mediastinal nodes (62 patients). Locoregional control for stages I, II, and IIIA was 85, 75, and 85%, respectively. Five-year actuarial survival was 35% for stage I and 20% for stages II and IIIA. Patients with N0 disease (positive margins) had 5-year survival of 25%, whereas patients with N1 and N2 disease had a 5-year survival of 20%. PMID- 9345327 TI - The role of chemotherapy in head and neck cancer. AB - We studied the effect of cytoreductive chemotherapy in head and neck cancer and analyzed it in terms of efficacy, remission rates, and duration, as well effect on survival. Single-agent chemotherapy, which formerly was used as a palliative therapy in recurrent and metastatic disease, had little affect on survival. More recently, multi-agent chemotherapy trials have shown significantly higher response rates, but this success has not translated into an added survival benefit. These findings led to the introduction of multi-agent chemotherapy into the induction (neoadjuvant) clinical setting. In these clinical circumstances, better objective response rates were found, particularly in the previously untreated patient. Although this therapy has resulted in better control of local disease, the impact on survival is not yet clear. Adjuvant chemotherapy is most useful in patients who have a high risk of relapse. Therapy appears to decrease its incidence, particularly at distant sites. Finally, chemoradiation trials have shown that this treatment provides a survival advantage, but at the cost of a significant increase in toxicity. PMID- 9345328 TI - Transplantation in patients with multiple myeloma: a multicenter comparative analysis of peripheral blood stem cell and allogeneic transplant. AB - We performed a multicenter comparative analysis of autologous peripheral blood stem cell transplantation (PBSCT) and allogeneic bone marrow transplantation (alloBMT) in multiple myeloma. Forty-eight consecutive patients received either PBSCT (24 patients) or alloBMT (24 patients) at one of three institutions in the study group. Preparatory regimens consisted of melphalan and total body irradiation (TBI) or melphalan alone in the PBSCT group. The alloBMT group received one of four regimens: cyclophosphamide and TBI; cyclophosphamide, VP-16 and 1,3-bis(2-chloroethyl)-1-nitrosourea (CVB); busulfan and cyclophosphamide (BU/CY) and total marrow irradiation (TMI); or melphalan and TBI. Procedure related mortality was 12.5% for the PBSCT group and 25% for the alloBMT group. With a median follow-up for survivors in the PBSCT and alloBMT groups of 11 months (range, 4-46) and 15 months (range, 2-84 months), respectively, there was no significant difference in median overall survival (33.5 versus 38.6 months, p = 0.7637) or event-free survival (16.7 versus 31 months, p = 0.8450). There was, however, a plateau in survival at 40% in the alloBMT group. No plateau in survival was seen in the PBSCT group. Clinical relapses occurred as late as 39 months posttransplant. Patients have survived up to 28 months postrelapse. PMID- 9345329 TI - Solitary extramedullary plasmacytoma five years after successful cardiac transplantation: case report and review of the literature. AB - We document the occurrence of a solitary extramedullary plasmacytoma (SEP) in a cardiac transplant patient. The diagnosis of plasma cell malignancy was confirmed by histopathologic and immunohistochemical examination of a nodular skin lesion. A complete systemic evaluation showed no evidence of metastatic disease. The patient was treated locally with radiation therapy (RT), but disseminated multiple myeloma developed 4 months after diagnosis. A variety of tumors have been reported to develop in the cardiac or renal transplant recipient, although plasma cell malignancies are rare. To our knowledge, this is the first reported case of an SEP in an organ transplant recipient. PMID- 9345330 TI - Skeletal response to clodronate in prostate cancer with bone metastases. AB - Bone metastases, together with generalized bone resorption, represent the main complication in patients with advanced prostate cancer, and palliative treatments are required to delay the progression of the metastases and improve the quality of life of these patients. For this reason, the bisphosphonate clodronate was administered to 18 patients (clodronate group) from a total of 30, all of whom were receiving complete androgenic blockade; the remaining 12 formed the control group. Transiliac bone biopsies were taken at the beginning of the study and 6 months later to determine the effect of the bisphosphonate on the skeleton. The results were assessed by bone histomorphometry and showed, although without statistical significance between the groups, an antiresorptive effect of the clodronate expressed as the eroded surface/bone surface and as the osteoclast number/bone surface. However, the bone volume also decreased after 6 months of treatment. Similarly, osteoid formation decreased as indicated by the osteoid surface and by the osteoid volume, probably due to the effect of the drug on the osteoblasts. The mineralization rate was apparently slightly retarded in the clodronate group, although to a lesser degree than in the control group. The results confirm the antiresorptive effect of clodronate and its detrimental effect on osteoblast activity. PMID- 9345331 TI - The clinical impact of teniposide in the treatment of elderly patients with small cell lung cancer. AB - Teniposide (VM26) has been claimed to be active with a moderate toxicity in elderly patients affected by small-cell lung cancer (SCLC). Twenty-two patients with SCLC older than 65 years received VM26 as first-line chemotherapy at a dose of 60 mg/m2 on 5 consecutive days every 3 weeks. Age distribution ranged from 67 to 80 years (median 72 years). Fourteen patients were men and eight were women. Twelve patients had limited disease (LD) and ten extensive disease (ED). One patient (LD) had a complete response, and four (3 LD, 1 ED) achieved a partial response for an overall response rate of 22.7% (95% CI 6-40%). The most frequent toxicity was myelosuppression: 20 and 15% of patients had grade 3 leukopenia and thrombocytopenia, respectively. Our results seem to suggest that VM26 by this schedule is moderately effective in elderly patients with SCLC, and it cannot be recommended as a routine treatment. PMID- 9345332 TI - The location of the prostatic apex on retrograde urethrography and its relationship to the bottom of the ischial tuberosities. AB - To determine the proportion of patients undertreated if the inferior border of the prostate field is set at the bottom of the ischial tuberosities, we reviewed the ports of 80 patients with prostate cancer who had retrograde urethrography as part of simulation for radiation therapy. For the 75 evaluable urethrograms, the mean distance from the top of the urethrogram cone to the bottom of ischial tuberosities was 1.38 cm (range, -0.48-2.90 cm). A comparison of the inferior border defined by the bottom of the ischial tuberosities and retrograde urethrography showed that 47 of 75 (62.7%) patients would have been undertreated if a margin of 1.5 cm was employed, and the prostatic apex was thought to be directly above the urethrogram cone. If the apex was thought to be 1 cm above the cone, six of 75 (8.0%) patients would have been undertreated, using a margin of 1.5 cm. Although previously published reports have established that using the bottom of the ischial tuberosities as the inferior border of the prostate field results in 10-45% undertreatment, our findings, when adjusted for the variability of prostatic apex location and margin of normal tissue employed, indicate that only 8% may actually be undertreated. PMID- 9345333 TI - Retrospective comparison of infusional 5-fluorouracil, doxorubicin, and mitomycin C (modified FAM) combination chemotherapy versus palliative therapy in treatment of advanced gastric cancer. AB - About one-third of patients with gastric cancer are unresectable at the time of diagnosis. Their median survival is < 6 months, with a grave prognosis. The purpose of this study was to assess the efficacy of a modified FAM (mFAM) regimen in advanced gastric cancer. We retrospectively reviewed the clinical records of 409 advanced gastric cancer patients who had not received curative surgery. Among 409 patients, 202 patients were treated with an mFAM regimen (infusional 5-FU + doxorubocin + mitomycin-C), and 207 patients received no chemotherapy (control group). No differences were found in clinical parameters between the two groups. The 1-year survival rates were 34.1% for the mFAM-treated group and 22.5% for the control group (p = 0.0135). In subset analysis, a higher 1-year survival rate was demonstrated in patients with mFAM and palliative surgery. Of the 154 evaluable patients in the mFAM-treated group, the response rate was 17.5%. In these patients, median response duration was 30 weeks, and progression-free survival was 23 weeks. Overall toxicity of mFAM regimen was relatively tolerable and reversible. In conclusion, FAM combination chemotherapy, which has been used as a standard therapy, prolonged survival after modification of the administration schedule and combination with palliative surgery. A prospective randomized study is warranted to confirm this conclusion from our retrospective study. PMID- 9345334 TI - Phase II study of combined levamisole with recombinant interleukin-2 in patients with advanced malignant melanoma. AB - Adoptive immunotherapy (AI) with interleukin-2 (IL-2) and lymphokine-activated killer cells (LAK) is an antineoplastic modality in which immune-activated cells are administered to a host with advanced cancer in an attempt to mediate tumor regression. Levamisole (LEV), an immune stimulant, has been suggested to have therapeutic effectiveness in a variety of cancers. After a phase I trial of recombinant IL-2 plus LEV, a phase II trial of this combination was conducted in patients with advanced malignant melanoma. Nineteen patients were entered in the trial. They received IL-2 at 3 x 10(6) U/m2 subcutaneously daily x 5 plus LEV 50 mg/ m2 orally three times daily (p.o. t.i.d.) x 5. Patients were reevaluated at four-week intervals. None of the patients achieved a partial or complete regression (PR, CR). The median time to treatment failure (refusal, progression, or off study due to toxicity) was 56 days. Grade IV toxicities included vomiting (3 patients), lethargy (1 patient), and musculoskellar pain (1 patient). This regimen is not recommended for further testing in patients with advanced malignant melanoma. PMID- 9345335 TI - Induction intraarterial chemotherapy for T4 breast cancer through an implantable port-catheter system. AB - Eleven patients with T4 breast cancer received induction intraarterial chemotherapy (IACT) as the first step in multidisciplinary therapy. The IACT agents (epirubicin and mitomycin C), were delivered weekly in the outpatient department by bolus injection through an implantable port-catheter system. A modified technique of port-catheter system implantation was used. The precise localization of the catheter was dually confirmed by angiography and dye test. The effectiveness of the treatment was evaluated by clinical appearance, image study, and microscopic examination. A 91% response rate was obtained, and the lesions were resectable in < or = 8 weeks. No obvious systemic toxicity resulted from the IACT. Our results show that weekly IACT by bolus injection through a port-catheter system for treating locally advanced T4 breast cancer is feasible and efficacious. PMID- 9345336 TI - Combination chemotherapy with cisplatin, carboplatin, and etoposide in advanced malignancy: a phase I trial. AB - Cisplatin and carboplatin are platinum-based chemotherapeutic agents with broad antitumor activity and significantly different toxicity profiles. They are commonly used in combination with etoposide (VP-16) in chemotherapeutic regimens. We conducted a phase I trial using the combination of cisplatin, carboplatin, and etoposide in advanced malignancy, aimed at delivering a higher dose intensity of active platinum species while taking advantage of their nonoverlapping toxicities. Etoposide was added because of its synergistic action with platinum compounds. The initial chemotherapy regimen consisted of carboplatin 180 mg/m2 on day 1, cisplatin 70 mg/m2 on day 1, and etoposide 60 mg/m2 on days 1-3. Dose was escalated based on toxicity observed at each level and separately for patients with a previous history of chemotherapy and for those with no prior treatment. Thirty-six patients were entered in the study, and 33 were evaluable. Hematologic toxicity was dose limiting. Grade 3-4 leukopenia was noted in 22 of 33 (66%) patients and grade 3-4 thrombocytopenia was noted in 16 of 33 (48%). No serious bleeding complications occurred. There was one treatment-related death due to neutropenic sepsis. Nonhematologic toxicity was mild and not dose limiting. Ototoxicity and nephrotoxicity were minimal. No complete responses (CR) occurred. Nine of 33 (27%) patients had objective responses, including 3 patients with adenocarcinoma of the esophagus or gastroesophageal junction who had failed prior chemotherapy. Fifteen of 33 (45%) patients had stable disease. The maximum tolerated dose varied for patients who had received prior chemotherapy and for those who were previously untreated. For further studies, the recommended dosing for previously untreated patients is carboplatin 300 mg/m2 on day 1, cisplatin 70 mg/m2 on day 1, and etoposide 105 mg/m2 on days 1-3. The recommended dosing for patients with a history of prior chemotherapy is carboplatin 220 mg/m2 on day 1, cisplatin 70 mg/m2 on day 1, and etoposide 75 mg/m2 on days 1-3. The combination of cisplatin, carboplatin, and etoposide merits further testing in phase II trials. PMID- 9345338 TI - Hepatocellular carcinoma presenting as a pancreatic head mass: report of an unusual case. AB - Hepatocellular carcinoma (HCC) spreading to the pancreas is rare and, when it does occur, is most often detected as a late finding at autopsy. We report the first case of HCC initially presenting as a pancreatic mass. Computed tomography (CT) scan showed a 7-cm mass in the dome of the liver and a 3-cm pancreatic head mass causing biliary and duodenal obstruction. Palliative surgery was performed to bypass the pancreatic obstruction. During the operation, fine needle aspiration (FNA) was performed on the pancreatic mass. The liver mass was sampled by percutaneous ultrasound-guided biopsy. Postoperatively, the patient's obstructive symptoms were relieved. The histologic studies showed that both the liver and the pancreatic masses consisted of poorly differentiated HCC that stained positive for alpha-fetoprotein. The rarity and the implications of this presentation are discussed. PMID- 9345337 TI - Miliary osteosarcomatosis with associated hypocalcemia. AB - We report the case of a patient with a rare miliary variant of osteosarcoma associated with symptomatic hypocalcemia and review the literature regarding the pathogenesis of multifocal osteosarcoma, treatment options, and the associated hypocalcemia. PMID- 9345340 TI - Mitomycin C, cisplatin, and 5-fluorouracil for advanced and/or recurrent head and neck squamous cell carcinomas. AB - The combination of cisplatin (CDDP 100 mg/m2 on day 1) and 5-fluorouracil (5-FU 1,000 mg/m2 continuous intravenous (i.v.) infusion days 1-5) is the most widely used chemotherapy regimen for the treatment of advanced head and neck carcinomas, with a response rate of 70-90% but with a survival and a duration of response which are not impressive. Most patients relapse in < or = 2 years and die of cancer. We evaluated the activity of a CDDP (90 mg/m2 on day 1), 5-FU (900 mg/m2/120 h continuous i.v. infusion from day 1), and mitomycin C (MMC 6 mg/m2 on day 1) regimen in advanced or recurrent head and neck squamous cell carcinoma (HNSCC). Fifty-six patients were treated and evaluated for response and toxicity: 5 (9%) complete responses (CR) and 36 (64%) partial responses, (PR) were observed (response rate 73%). The median duration of response was 12 months, and median survival was 15 months. At a median follow-up of 14 months, the estimated overall survival at 1 year was 65%; at 2 years, it was 35%. Grade 3-4 toxicity was noted in 14 patients, mostly hematologic; overall toxicity required a dose-intensity decrease in 20.2% of all cycles. No treatment-related deaths occurred. The regimen showed a good response rate and an encouraging median duration of response with a good tolerability profile. PMID- 9345339 TI - Phase I study combining tumor necrosis factor with interferon-alpha and interleukin-2. AB - We evaluated the effects of the addition of escalating doses of tumor necrosis factor (TNF) to two fixed doses and schedules of a combination of interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) to determine the maximum tolerated dose of this three-cytokine combination and its feasibility as an outpatient regimen. Eighteen patients with metastatic cancer were enrolled. Each course consisted of 3 consecutive weeks of treatment with IFN-alpha 9 x 10(6) IU/m2/day intramuscularly (i.m.) or subcutaneously (s.c.) days 1, 3, and 5 each week for 3 weeks plus IL-2 continuous infusion 1 x 10(6) IU/m2/day (group A) or 3 x 10(6) IU/m2/day (group B) days 1-5 each week for 3 weeks. TNF was administered only during the first week of each course intravenously (i.v.) for 2 h on days 1-5. The dose of TNF was escalated (40, 80, 120 micrograms/m2) in cohorts of 3 patients. The most common side effects were fever, chills, and fatigue in all patients. Grade 3-4 toxicity included anemia (3 patients), thrombocytopenia (1 patients), arrhythmia (2 patients), pulmonary edema (3 patients),- and weight loss (1 patient). Five patients withdrew from study due to toxicity. The combination of the three cytokines is feasible as an outpatient regimen in one of the following combinations: (a) TNF 80 micrograms/m2/day as 2-h infusion on days 1-5 + IL-2 1 x 10(6) IU/m2/day continuous infusion on days 1-5 for 3 weeks + IFN alpha 9 x 10(6) IU/m2/day s.c. or i.m. on days 1, 3, and 5 for 3 weeks, or (b) TNF 40 micrograms/m2/day as a 2-h infusion on days 1-5 + IL-2 3 x 10(6) IU/m2/day continuous infusion on days 1-5 for 3 weeks + IFN-alpha 9 x 10(6) IU/m2/day s.c. or i.m. on days 1, 3, and 5 for 3 weeks. PMID- 9345341 TI - Second line chemotherapy with ifosfamide as outpatient treatment for advanced bladder cancer. AB - We have carried out a phase II study in advanced or metastatic transitional cell carcinoma of the bladder. Eligible patients had unresectable bladder cancer, previously treated with one line of systemic chemotherapy. Treatment consisted of ifosfamide 1000 mg/sm in a 2-hour infusion for 5 consecutive days from d.1 to d.5. Mesna was administered intravenously at a 20% of the ifosfamide dosage before ifosfamide and orally at 40% after 4 and 8 hours from the ifosfamide infusion. Twenty patients entered the study and received a total of 62 cycles: the treatment resulted feasible on an outpatient basis, with mild toxicity. Only one partial response was observed. With this dose and schedule, ifosfamide appeared less effective than in a previous report at higher doses. Toxicity was acceptable. PMID- 9345342 TI - Long-term results in patients with advanced epithelial ovarian carcinoma treated with a combination of cisplatin, doxorubicin, and cyclophosphamide. AB - From 1984 to 1988, thirty-nine untreated patients with epithelial ovarian cancer received Cisplatin 50 mg/m2, Doxorubicin 50 mg/m2, and Cyclophosphamide 750 mg/m2 (CAP), at 3 weekly intervals. All patients had FIGO stage III or IV tumors except 2 patients with stage IIb and IIc, respectively. After initial surgery 23 patients had residual disease > 2 cm in diameter. Twenty-five patients (64%) were evaluable for response to chemotherapy. Objective responses were observed in 13 out of 25 patients (52%, 95% confidence intervals (CI), 32.42% to 71.58%), 6 patients had a cCR (24%) and 7 had a cPR (28%). Seventeen out of the 39 patients (44%) had a second-look laparotomy. A pCR was achieved in 5 out of 17 patients (29%); a pPR was obtained in 8 patients (47%). Median duration of survival was 41,5 months (range 2-107+); median duration of time to failure was 21 months (range 2-107+). Median disease-free survival was 86 months (range 3,5-107+). Eleven patients (28%) are alive and 9 patients (23%) are free of recurrence at median follow-up of 86 months. Only 4 of 11 long-term survivors had a pCR. In univariate analysis, histology, clinical response to chemotherapy, and the presence of ascites at the time of diagnosis, achieved a significant correlation with survival and time to failure (TTF); in addition, TTF was significantly affected by pathological response to induction chemotherapy. The only important predictors of disease-free survival (DFS) were tumor grade and stage of disease. In multivariate analysis, the presence of ascites was the only significant prognostic factor with respect to survival and TTF. Our study confirms the effectiveness of CAP in the treatment of epithelial ovarian cancer and the relatively poor long term prognosis. PMID- 9345344 TI - American Radium Society, 1916-1995: years of distinction. Hartman Centennial Lecture. 77th annual American Radium Society meeting. Paris, France. May 1, 1995. PMID- 9345343 TI - Irinotecan hydrochloride (CPT-11) resistance identified by K-ras mutation in patients with progressive colon cancer after treatment with 5-fluorouracil (5 FU). AB - OBJECTIVE: To determine the prognostic role of a K-ras mutation in tumor tissue of patients with refractory colon cancer who received irinotecan hydrochloride (CPT-11). METHODS: DNA was extracted from paraffin-stored tumor tissue of 35 patients with progressive colon cancer failing treatment with 5-fluorouracil who subsequently received CPT-11 (100 mg/m2 i.v. per week x 4 weeks with 2 weeks off per course). The first exon of the K-ras gene was amplified by polymerase chain reaction by using K-ras-specific primers followed by mutant enrichment sequencing. Survival differences of patients with a K-ras mutation were compared with those of patients with a normal K-ras status. RESULTS: A total of 21 patients had a normal K-ras sequence and 14 patients had a K-ras mutation [GAT, n = 7; TGT, n = 3; and GCT, AGT, GTT, GAC (codon 13), n = 1 each]. Median survival of patients with a normal ras sequence from time of treatment with CPT-11 was 332 days compared with 169 days for patients with a K-ras mutation (p = 0.0036). No differences in age, sex, cancer stage, surgical treatment, or chemotherapy treatment were observed. CONCLUSION: Determination of the presence of a K-ras mutation may predict survival in patients with progressive colon cancer after treatment with 5-fluorouracil who receive CPT-11. PMID- 9345345 TI - Breast cancer: a systemic or local disease? AB - Since ancient times, controversy has centered on whether breast cancer is a systemic or local disease at inception. In recent years, support for the systemic hypothesis has come from randomized clinical trials showing that variations in locoregional therapy have no impact on breast cancer mortality. Yet at first glance, the results of the breast cancer screening trials seem to refute this hypothesis. These trials indicate that screening reduces breast cancer mortality by 30% in postmenopausal women. This could be interpreted to mean that 30% of postmenopausal breast cancers metastasize relatively late in their natural history and are therefore amenable to cure with early diagnosis and timely extirpation. Closer scrutiny of the results of the screening trials, however, shows that screening changes the slope of the breast cancer survival curves (so that survival is prolonged) but does not cause the curves to plateau. Thus, screening may simply increase the time to recurrence and death, rather than eliminate the risk of death for a fraction of postmenopausal women with breast cancer. One may speculate that the timely extirpation of the primary tumor reduces the burden of micrometastatic disease, allowing the host's own defense to exert an effect. Alternatively, the benefit of screening is perhaps the result of early administration of systemic therapy, when the burden of micrometastatic disease is low. Regardless, the results of the screening trials are not necessarily inconsistent with the systemic hypothesis. Longer follow-up of women enrolled in the screening trials may prove useful in better understanding the natural history of breast cancer. PMID- 9345346 TI - Taxol-induced cellulitis after extravasation: a rarely reported event. PMID- 9345347 TI - When one more Pap smear is one too many. PMID- 9345348 TI - The value of repeat Pap smear at the time of initial colposcopy. AB - OBJECTIVE: The objective was to determine if repeating a Pap smear at the time of an initial colposcopy has sufficient clinical benefit to justify its clinical and financial costs. METHODS: The records were reviewed of all patients who had an initial colposcopy at Queens Hospital Center between 1984 and 1995. Data were gathered regarding the referral cytology, the cytology done at the time of colposcopy, and the results of any biopsies which were taken. The terminology for cytology and histology done prior to 1989 was adjusted to the Bethesda classification system. A repeat Pap smear was defined as clinically valuable if it would have changed the patient's management, i.e., if it suggested more advanced disease than the referral Pap and that the disease was not identified on the colposcopically directed biopsy. RESULTS: Two thousand nine hundred sixty nine records were reviewed. In 139 cases, no Pap smear was repeated at the time of colposcopy. Of the remaining 2830 women, only 1347 (47.6%) showed exact correlation between their referral Pap smear and the Pap done at the time of colposcopy. In another 1016 (35.9%), the Pap at colposcopy was within one grade of the referral Pap. In 312 women, the Pap at the time of colposcopy was a higher grade than the referral Pap. However, in 236, the higher grade of disease was detected by the colposcopically directed biopsy. Of the remaining 76 women, 58 had a normal biopsy, but their Pap at the time of colposcopy showed low-grade squamous intraepithelial lesions (44) or high-grade squamous intraepithelial lesions (HGSIL) (14). Seventeen others had a biopsy showing low-grade dysplasia while the Pap at the time of colposcopy showed HGSIL. In 1 patient, the repeat Pap showed malignant cells while the biopsy showed a high-grade lesion. Based on the triage protocols at our institution, this means that a repeat Pap at the time of colposcopy would have indicated a cone biopsy in 31 patients (1.1%) and more careful follow-up of another 44 patients (1.6%). Skipping the repeat Pap smear would not have resulted in any missed cancers. In our series of 2830 patients, the cost savings of skipping the repeat smear would have been $68,580 or $24.23 per patient. On a national level, skipping the repeat smear would save more than $24,000,000 annually. CONCLUSION: Using current triage protocols at our institution, repeating the Pap smear at the time of an initial colposcopy would have changed the management in 2.7% of patients and indicated a conization in only 1.1% of patients. It is doubtful that this justifies its cost and the potential detrimental effects on the colposcopic examination. PMID- 9345349 TI - Estrogen and progesterone receptor expression in postmenopausal tamoxifen-exposed endometrial pathologies. AB - Assessment of receptor levels in tamoxifen-exposed endometrial pathologies may indicate endometrial cells potential for interaction with tamoxifen. To assess this assumption, we analyzed estrogen receptor (ER) and progesterone receptor (PR) expression by an immunohistochemical technique in endometrial specimens with benign hyperplasia, benign polyps, and carcinoma obtained from postmenopausal breast cancer patients treated with tamoxifen (study group) and from age-matched healthy postmenopausal women treated with estrogen replacement therapy (control group I) and not treated with estrogen replacement therapy (control group II). Overall gland and stromal ER expression of benign endometrial hyperplasia and of benign endometrial polyps was significantly higher in control groups I and II than that obtained from the study group (endometrial hyperplasia: P = 0.0274 and 0.00093, respectively, and P = 0.00003 and 0.00001, respectively; benign endometrial polyps: P = 0.02889 and 0.00596, respectively; and P = 0.00228 and 0.00005, respectively), while there were no differences between the two control groups. Overall gland and stromal PR expression was nearly similar in all the three groups (P = NS). There was no correlation between the length of tamoxifen treatment and the presence of ER and PR in various endometrial pathologies in the tamoxifen-treated patients. The significantly lower ER expression in most benign endometrial pathologies obtained from postmenopausal tamoxifen treated patients may further support the weak estrogen-like effect of tamoxifen on the endometrium in the menopause. PMID- 9345350 TI - The accuracy of frozen section by tumor weight for ovarian epithelial neoplasms. AB - This study evaluated the effect ovarian weight has on the accuracy of frozen sections in serous and mucinous ovarian tumors. The study group included 294 patients who had an initial frozen section (189 serous and 105 mucinous tumors) at surgery. The pathology reports were separated into subgroups (benign, borderline, or malignant). Tumors were broken down into three weight categories: < or = 450 g, > 450 to < or = 1360 g, and > 1360 g. In each weight category, accuracy, sensitivity, specificity, and positive and negative predicative values were calculated on frozen sections. The mean weight of the ovarian tumors was 1042 g. As the weight increased in serous tumors, the sensitivity fell from 96.2 to 93.8 to 75%, respectively, in each weight category. The same trend was noted with mucinous tumors as sensitivity fell from 91.7 to 87.5 to 66.7%, respectively. With an increase in the size of ovarian tumors, a decrease in the sensitivity of frozen section was observed. With tumors greater than 1360 g, sensitivity was only 69%. Twenty-three percent of ovarian tumors revealing borderline diagnosis at frozen section were malignant on the final pathology report, with the greatest misclassification in > 1360-g mucinous tumors (50%). For patients with large ovarian tumors, consideration should be given to performing staging at the time of the initial laparotomy. PMID- 9345351 TI - The relevance of angiogenesis in benign and malignant epithelial tumors of the ovary: a quantitative histologic study. AB - The ability of a tumor to grow and eventually to infiltrate adjacent tissues requires a sufficient blood supply. Many malignant neoplasms have been shown to induce neovascularization. We asked whether it is possible to characterize ovarian epithelial tumors on the basis of vascularization and whether vascularization can be used to differentiate between benign and malignant neoplasms. We examined 14 cases of benign cystadenomas and 18 carcinomas. The microvessels were identified by immunohistochemical staining of endothelial cells for factor VIII. They were counted within the most vascular area of a tumor (neovascular "hot spot") on a x 100 and a x 400 field. Mean vessel counts were 51.64 [standard error of the mean (SEM), 5.7] in the benign cystadenomas and 131.05 (SEM, 6.7) in the group of carcinomas at a x 100 magnification. The mean microvessel counts per x 400 field were 14.4 (SEM, 1.9) in the benign and 33.7 (SEM, 3.45) in the malignant tumors investigated. These differences were significant (t test for independent samples, P < 0.001). Since our study shows significantly fewer small blood vessels in the benign cystadenomas than in the malignant tumors, the histologically determinable vascular density may be the basis for imaging blood flow by means of color ultrasound. PMID- 9345352 TI - Angiogenesis in cervical neoplasia: microvessel quantitation in precancerous lesions and invasive carcinomas with clinicopathological correlations. AB - Recently, angiogenic properties have been shown in preinvasive cervical lesions. Our goal was to determine the angiogenesis in cervical intraepithelial neoplasia (CIN) and the relationship between microvessel counts, histopathological parameters, and clinical outcome in invasive cervical carcinoma. One hundred thirty-eight cervical specimens were evaluated; among these 20 were designated normal epithelium, 20 low-grade CIN, 40 high-grade CIN, and 58 invasive carcinoma. Histological sections immunostained for CD31 were quantitatively evaluated for microvessel density. The tumor proliferation rate was determined by the Ki-67 Labeling Index. Comparison of microvessel counts from normal epithelium with those from CIN and invasive carcinoma showed significant increases in precancerous lesions and invasive cancer (P < 0.0001). Microvessel density was found to be associated with the overall survival in women with invasive carcinoma (P < 0.01). There was a significant correlation of microvessel density (P < 0.05) with relapse-free survival in patients with regional lymph node metastasis. A Cox stepwise regression analysis revealed microvessel density, together with depth of invasion, regional lymph node status, and vascular invasion, to be a strong independent prognostic indicator for overall survival in patients with clinical stage IB cervical carcinoma. PMID- 9345353 TI - Outcomes after cervical cold knife conization with complete and incomplete excision of abnormal epithelium: a review of 699 cases. AB - A retrospective analysis was undertaken of 699 cone biopsies performed at the Royal Women's Hospital in Melbourne from 1966 to 1992. In 572 cases (82%), abnormal epithelium was assessed as having been completely excised, and in 127 (18%) excision was incomplete. There were no significant differences in age, parity, cytology, histology, or indications for conization between patients in whom excision was incomplete and those in whom complete excision was achieved. Of the patients whose cone biopsy histology showed complete excision of abnormal epithelium, 96.7% were found to have been cured of disease on the basis of normal follow-up or normal histology of hysterectomy specimens. The overall cure rate after incomplete cone biopsy was found to be 77%, but was influenced by the site of incomplete excision. The cure rate for incomplete excision at the ectocervical margin was 86%; incomplete excision at the endocervix was 68% and only 40% if excision was incomplete at both edges. Cone biopsy undertaken for cervical intraepithelial neoplasia is likely to be curative when the lesion is completely excised, but recurrent disease may occur and adequate follow-up is an essential part of patient management after conization, regardless of histological findings in the conization specimen. Most cases of incompletely excised cervical intraepithelial neoplasia will also be cured, especially if the incomplete margin is ectocervical, and cytological and colposcopic follow-up may be an acceptable alternative to repeat cone biopsy or hysterectomy in the management of such cases. PMID- 9345355 TI - Epidermal growth factor activates protein kinase C in the human endometrial cancer cell line HEC-1-A. AB - Previous studies from this laboratory have shown that epidermal growth factor (EGF) and the tumor promoter, phorbol myristate acetate (PMA), are mitogenic in the endometrial cancer cell line HEC-1-A. Since the effects of EGF have been shown to be mediated by the protein kinase C (PKC) pathway transduction system, we examined the possibility that the EGF-responsive signal in the endometrial cancer cell line HEC-1-A involves protein kinase C activation. HEC-1-A cells were grown to confluency in 100-mm dishes and maintained in a serum-free medium for 24 hr prior to treatment. The cells were treated with EGF at varying time intervals (0.25 to 60 min) and concentrations (0.1 to 200 ng/ml). The cells were then lysed, homogenized, and centrifuged at 105,000g for 1 hr at 4 degrees C. The supernatant was chromatographed on DEAE-Sephacel columns. The membranous pellet was resuspended in 5 ml of lysis buffer containing 1% Nonidet P-40 and also chromatographed on DEAE-Sephacel columns separately. The eluates were collected and assayed for protein kinase C activity by determining the amount of 32P transferred from [gamma-32P]ATP onto histones in the presence of the phospholipids, phosphatidylserine, and diolein. Our results show that the cytoplasmic and membrane fraction of the HEC-1-A cell line contained phosphotransferase activity which displayed kinetic characteristics typical of the protein kinase C enzyme. The optimal incubation time for protein kinase C activation in the cytosol by EGF was 5 min (30-fold stimulation). The protein kinase C activity was increased when the cell lines were incubated with increasing concentrations of EGF. Enzyme saturation was seen at a concentration of 10 ng/ml of EGF (4.5-fold stimulation). Western blot analysis confirmed the presence of the PKC enzyme in the cytosol and membranes of our cancer cell line. These results suggest that EGF, at least partially, exerts its effects on the endometrial adenocarcinoma cell line by activating protein kinase C through increased breakdown of phosphatidyl inositol (PI). The PI cascade appears to be an important signal transduction system mediating the growth stimulatory effects of EGF on endometrial carcinoma. PMID- 9345354 TI - High-intensity intravenous cyclophosphamide and cisplatin, interim surgical debulking, and intraperitoneal cisplatin in advanced ovarian carcinoma: a pilot trial with ten-year follow-up. AB - PURPOSE: This trial was undertaken to study the effect of intensified intravenous cyclophosphamide/cisplatin and interim surgical debulking, followed by intraperitoneal cisplatin on surgically defined complete remission rate and survival in advanced ovarian cancer. PATIENTS AND METHODS: Forty patients with stage IIB through IV ovarian cancer were entered and 36 were evaluable for response and survival and approximately 10 years. Following a first laparotomy for diagnosis and debulking, the patients received two cycles, spaced 28 days apart, of intravenous cisplatin 30-40 mg/m2/day with hypertonic saline for 4 to 5 days and cyclophosphamide 200 mg/m2/day for 5 days. A second laparotomy was done to further debulk remaining cancer and to place an intraperitoneal catheter. Four cycles of intraperitoneal cisplatin at 50 or 100 mg/m2 were administered 21 days apart and followed by a third laparotomy to define response and plan any further therapy. RESULTS: The surgically confirmed complete response rate was 47% and median survival is 68.3 months for this group. Ten of the 17 patients (58.8%) relapsed following complete response at a median of 19.5 months (range, 5-98). Both aggressive chemotherapy and surgery seemed to play a role in inducing this high complete response rate. Traditional prognostic factors, including stage and diameter of largest residual disease, had little apparent effect on likelihood of complete response or survival, whereas tumor grade had a more significant effect on survival. Nadir fever was experienced by 33% of patients but peripheral neuropathy was dose limiting. CONCLUSION: In the context of recent data failing to support any clinical benefit to modest increases in dose escalations of cisplatin or carboplatin, in this trial the high complete response rate suggests that the multimodality approach (i.e., interval surgical debulking and intraperitoneal cisplatin) is worthy of further study. The high relapse rate among complete responders and the unacceptable neurotoxicity also suggest that modifications could improve the results. The use of newer agents and further intensification (substituting carboplatin for cisplatin and the use of paclitaxel) with stem cell support are two examples. PMID- 9345356 TI - Incidence of atypical glandular cells of uncertain significance in cervical cytology following introduction of the Bethesda System. AB - OBJECTIVE: To establish the frequency of the atypical glandular cells of uncertain significance (AGCUS) category, and its subcategories, as defined by the Bethesda System (TBS). METHODS: Our computerized records of cervical/vaginal cytology specimens submitted from January 1, 1993, through December 31, 1995, were retrospectively reviewed for specimens diagnosed as AGCUS. When appropriate, our subcategory of "AGCUS favor premalignant/malignant lesion" was further qualified as "favor endocervical adenocarcinoma in situ" or "suspicious for endometrial carcinoma." The number of specimens and patients diagnosed for each subcategory were grouped by calendar year. Differences in frequency between time periods were tested for statistical significance using chi 2 analysis. RESULTS: AGCUS was diagnosed in 1181 of 177,715 submitted specimens (0.66%). The frequency of subcategories was as follows: "favor reactive" (65%), "unable to further classify" (30%), "favor premalignant/malignant" (2.9%), "suspicious for endometrial carcinoma" (1.9%), and "favor endocervical adenocarcinoma in situ" (0.4%). From 1993 to 1995 there was an increase in the rate of diagnosis of AGCUS (0.55 to 0.73%; P < 0.001) and a decrease in the percentage of specimens with AGCUS subclassified as "favor premalignant/malignant" (6.2 to 0.5%; P < 0.001). Other subcategories showed no significant change in frequency over this time period. The rate of biopsy-proven preinvasive or invasive lesions in AGCUS patients also showed no significant change from year to year over this time period. CONCLUSION: The AGCUS diagnosis can be anticipated at a low but consistent rate from a cytology laboratory using TBS. Any comparison of laboratories should take into consideration the change in reporting frequencies that occurs as part of the "learning curve" following introduction of TBS reporting. Uniform diagnostic criteria and additional reports with large numbers of cytologic specimens will be needed to establish the expected frequency of AGCUS and its subcategories. PMID- 9345357 TI - The role of surgical cytoreduction in Stage IV endometrial carcinoma. AB - OBJECTIVE: To evaluate the impact of surgical cytoreduction in patients (pts) with Stage IV endometrial adenocarcinoma. METHODS: We performed a retrospective chart review of all pts with Stage IV endometrial carcinoma treated at our institution from January, 1977, to February, 1995. RESULTS: Fifty-five patients who underwent surgery as part of their primary treatment for Stage IV endometrial carcinoma were identified. They were divided into three groups: Group I consisted of 24 pts (44%) who underwent optimal surgical cytoreduction (diameter of largest residual tumor nodule < or = 2 cm); Group II contained 21 pts (38%) who underwent suboptimal surgical cytoreduction (> 2 cm residual disease); Group III consisted of 10 pts (18%) who had unresectable carcinomatosis and had no cytoreduction at all. There were no statistically significant differences between the three groups with respect to median age at diagnosis, tumor grade, histologic subtype, or the presence of extra-abdominal metastases. The median survival rates for the three groups were I, 31 months; II, 12 months; and III, 3 months (P < 0.01). Within Group I, there was no statistically significant difference in survival between the 8 pts who were found at laparotomy to have metastatic disease < or = 2 cm and the 16 pts who initially had metastatic disease > 2 cm and were subsequently cytoreduced to optimal status. On multivariate analysis only the extent of surgical cytoreduction had prognostic significance on survival. CONCLUSION: The prognosis of Stage IV endometrial carcinoma is poor. However, the data in this series suggest that aggressive tumor cytoreduction may improve survival in these patients. PMID- 9345359 TI - Carcinosarcoma of the uterus: a clinicopathological multicenter CTF study. AB - The purpose of this paper is to retrospectively analyze the clinical and pathological data of 118 cases of uterine carcinosarcoma. Prognostic factors and management were evaluated. chi 2 and log-rank tests were performed. Surgical stage at time of surgery was the most important prognostic factor (P < 0.0001); in stage I-II disease depth of myometrial invasion and lymphatic/vascular space involvement were significantly related to outcome, whereas other factors were not useful. In stage I-II patients postoperative radiation did not improve 5-year disease-free survival. Survival curves in patients with advanced disease treated with cisplatin-containing regimens or with doxorubicin (without cisplatin) containing regimens were overlapping. PMID- 9345358 TI - Long-term follow-up of the first randomized trial using neoadjuvant chemotherapy in stage Ib squamous carcinoma of the cervix: the final results. AB - OBJECTIVE: To determine if three courses consisting of 50 mg/m2 cis-platinum, 1 mg/m2 vincristine, and 25 mg/m2 bleomycin (day 1-3) at 10-day intervals can improve survival before Wertheim-Meigs + radiotherapy. MATERIAL: Two hundred five unselected stage Ib patients (having tumors > 2 cm in diameter) were divided into two groups at random: (1) The group control consisted of 103 patients (56 bulky, > 4 cm diameter) treated with Wertheim-Meigs (if the tumor was resectable with free surgical margins) + adjuvant radiotherapy to whole pelvis (extended field radiation was used only in patients with paraaortic lymph node metastases). When the tumor was unresectable, a surgical staging was performed and radiotherapy was the chosen treatment. (2) Neoadjuvant (102 patients, 61 bulky) had neoadjuvant chemotherapy and then the same treatment as the control patients. RESULTS: After 67 (31-102) months of follow-up, no difference was seen in tumors > 2 and < 4 cm in both groups (C = 77% vs N = 82%), but statistically significant differences were seen in survival and disease-free survival, in bulky tumors, and between patients with neoadjuvant chemotherapy + Wertheim-Meigs + radiotherapy (80%) and the control (61%). This was due to an increased operability that was substantially improved in bulky tumors in the neoadjuvant chemotherapy group (61/61, 100%) vs control (48/56, 85%; P < 0.01). After 7 years of follow-up, the outcome of the unresectable bulky control group of patients is significantly worse (14%) than that of the resectable group (69%; P < 0.001). With regard to recurrences, a significant decrease in pelvic failures in the neoadjuvant chemotherapy group was observed (P < 0.001). Survival was improved in bulky resectable cases (N = 81% vs C = 69%, P < 0.05). Pathological findings for the surgical specimens revealed differences between both groups because all the risk factors such as parametrial and lymph node metastases, tumor bulk, and vascular embolism had been decreased (P < 0.001). CONCLUSION: Neoadjuvant chemotherapy can improve survival because of increased operability with free survival margins and a decrease in pathologic risk factors in unselected, bulky (> 4 cm diameter) stage Ib patients. PMID- 9345360 TI - Fibroblasts stimulate human ovarian cancer cell invasion and expression of 72-kDa gelatinase A (MMP-2). AB - OBJECTIVE: The host-tumor interactions and tumor stroma may participate in the regulation of invasive behavior of tumor cells. In order to better understand the human ovarian cancer invasion we explored the possibility that normal fibroblasts could participate in the control of the spread of human ovarian cancer. RESULTS: A 3.5-fold increase (from 2.83 +/- 0.97 to 10.2 +/- 3.43%) in human ovarian cancer cell (Ovcar-3) invasion through a reconstituted basement membrane was noted when normal fibroblasts (CRL 1295) were added to the invasion chambers in conjunction with tumor cells. Conditioned medium from either fibroblasts or Ovcar 3 also enhanced the in vitro invasion of Ovcar-3 by 2- to 2.5-fold (from 2.83 +/- 0.97 to 5.71 +/- 3.5 and to 7.15 +/- 1.2%, respectively) compared to nonstimulated control cells. Zymographic analysis and assays of mRNA for the 72 kDa matrix metalloproteinase (MMP-2) showed that Ovcar-3 cells alone produced very low levels of MMP-2; the expression of this gelatinase was detectable in zymography only with stimulation by incubation of these cells with conditioned media from either fibroblasts or ovarian cancer cells themselves. Interestingly, MMP-2 activity was increased also in fibroblasts when using either ovarian cancer cell-conditioned (to 178 +/- 67%) or fibroblast-conditioned medium (to 215 +/- 61%) and the gene expression for MMP-2 was similarly increased in both fibroblasts and Ovcar-3 cells when using either fibroblast-conditioned medium or ovarian cancer cell-conditioned medium from 1.00 +/- 0.25 to 2.20 +/- 0.50 and 1.86 +/- 0.10 in fibroblasts and from 1.00 +/- 0.26 to 1.60 +/- 0.34 and 2.15 +/- 0.30 in Ovcar-3 cells, respectively. CONCLUSIONS: These results show that interplay between tumor cells and normal cells in the control of invasion and secretion of proteolytic enzymes may involve not only paracrine but also autocrine elements. Thus, such interactions are possible and may play an important role in the spread of cancer. PMID- 9345361 TI - Accuracy and safety of laparoscopic lymphadenectomy: an experimental prospective randomized study. AB - INTRODUCTION: The goal of this study was to investigate the accuracy and safety of bilateral pelvic and paraaortic lymphadenectomy performed via transperitoneal laparoscopy (LS) compared to laparotomy (LT) in a porcine model. MATERIALS AND METHODS: Fifteen adult, female hogs underwent LS and 15 underwent LT. A complete pelvic and paraaortic lymphadenectomy was performed in each animal by an experienced surgeon. Lymph nodes were counted by a pathologist in each case. Operative times were reviewed and included all procedures performed. The intraoperative complications were noted. Four weeks after the lymphadenectomy, the animals underwent exploratory laparotomy, and intraperitoneal adhesions were quantified. RESULTS: Thirty animals were evaluable. The average total number of lymph nodes retrieved by LS was 16.9 +/- 3.8, which was not statistically (P = 0.77) different from 16.5 +/- 4.9 nodes in LT. The average operating time in LT was 60 +/- 16 min compared with 128 +/- 24 min in LS. Twenty-eight animals were evaluable for adhesion formation. The average adhesion scores observed in anterior abdominal wall (P = 0.0006), paraaortic (P = 0.0005), right (P = 0.015), and left (P = 0.0324) iliac areas after LS were uniformly lower than after LT. DISCUSSION: This study indicates that laparoscopic pelvic and paraaortic lymphadenectomy is a safe and effective procedure. The node yield is similar for both approaches. The transperitoneal laparoscopy pelvic and paraaortic lymphadenectomy may not induce the degree of adhesion formation associated with laparotomy. PMID- 9345362 TI - Second-look laparotomy for ovarian cancer provides reliable prognostic information and improves survival. AB - From 1990 to 1995, 120 consecutive patients with stage IIIC and IV ovarian carcinoma underwent surgical cytoreduction to < or = 1-cm residual disease followed by platinum-based chemotherapy. At the conclusion of chemotherapy all patients who were clinically disease free and whose CA-125 was < 35 were offered a second-look operation that obtained at least 100 tissue specimens. Of 107 patients who qualified for second look, 78 underwent the procedure. Forty-three (55.1%) had negative pathology, 20 (25.6%) were microscopically positive, and 15 (19.2%) had gross disease. Patients with positive findings received individualized salvage therapy. Patient age (P = 0.01) and the number of implants at primary surgery (P = 0.004) correlated with second-look results. Twelve (27.9%) of the patients with negative pathology have recurred. Eleven of these patients had metastatic disease > or = 10 cm at primary surgery (P = 0.003). Patients refusing second look had a median survival of 39.1 months. Approximately 60% of patients who underwent second look remain alive. Stepwise logistic regression selected two covariates significantly affecting survival: the number of implants at primary surgery (P = 0.0130) and performance of a second look (P = 0.0103). Using the protocol described in a population of optimally resected patients with advanced-stage ovarian cancer, second-look laparotomy can impact positively on survival. Patients with > 10-cm metastatic disease at primary surgery and negative second-look findings should be the focus of future protocols for consolidation chemotherapy. PMID- 9345363 TI - Pure embryonal rhabdomyosarcoma of the fallopian tube. AB - Rhabdomyosarcoma is a neoplasm of childhood which commonly arises in the genitourinary tract. Reported locations include the bladder, prostate, paratestis, vagina, uterus, cervix, and ovary. Rhabdomyosarcomas have been reported to occur in the fallopian tube only as a component of a malignant mixed mullerian tumor. We present a case of pure embryonal rhabdomyosarcoma of the fallopian tube in a 17-year-old. The diagnosis was confirmed by immunohistochemical stains. The strongest evidence for the primary location of this pure embryonal rhabdomyosarcoma was the gross appearance of the tumor at laparotomy. Additionally, rhabdomyosarcomas arising from adjacent organs have never been reported to grow into the fallopian tubes. PMID- 9345364 TI - Cervical sarcoma botryoides and ovarian Sertoli-Leydig cell tumor. AB - The case of a woman who developed a cervical sarcoma botryoides tumor at age 14 years and a right ovarian Sertoli-Leydig cell tumor with alpha-fetoprotein production at 27 years is presented. The sarcoma botryoides was a stage 1b, 4-cm, polypoid ectocervical tumor treated by radical hysterectomy and bilateral pelvic lymphadenectomy. The Sertoli-Leydig cell tumor was a stage 1a, 145-g mass removed piecemeal by right oophorectomy. Histologically, it was an intermediate Sertoli Leydig cell tumor with a heterologous element composed of an endometrioid-like yolk sac tumor which was producing alpha-fetoprotein. There was no histological similarity between the two tumors. The patient is alive without evidence of disease, 16 years after diagnosis of her sarcoma botryoides and 3 years after her Sertoli-Leydig cell tumor. This is, to our knowledge, the third known association between these two rare gynecological tumors. The basis of the association remains unknown. PMID- 9345365 TI - Squamous cell carcinoma of the cervix metastatic to the spleen--case report. AB - Squamous cell carcinoma of the cervix metastatic to the spleen is an uncommon occurrence which has only been reported in small numbers in autopsy series. We present a case of a 47-year-old patient with a Stage IIb carcinoma of the cervix, treated with radiotherapy in 1990. Four years after completion of primary treatment she presented with a voluminous left hypochondrium and epigastrium mass. An exploratory laparotomy was performed and a splenic cyst 19 cm in diameter was found. The pathological examination revealed metastatic squamous cell carcinoma of the cervix in the spleen. Peritoneal washings were positive for malignant cells, due to incidental rupture of the cyst capsule. The patient received six courses of chemotherapy with a palliative intent and is alive, without further evidence of disease, 15 months posttreatment. To our knowledge this is the only case reported in the literature of squamous cell carcinoma of the uterine cervix metastatic to the spleen, diagnosed clinically as the only site of distant spread. PMID- 9345366 TI - Respiratory failure from metastatic choriocarcinoma: a survivor of mechanical ventilation. AB - A patient with respiratory failure from metastatic choriocarcinoma was treated with mechanical ventilation while receiving chemotherapy with etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine. The patient recovered from respiratory failure with the assistance of standard mechanical ventilation using low tidal volumes. The patient has sustained clinical remission with normal respiratory function. Mechanical ventilation with low tidal volumes and a pressure-targeted approach should be considered in the patient who develops early respiratory failure from metastatic choriocarcinoma. PMID- 9345367 TI - Epidemiology of cervical cancer in Taiwan. PMID- 9345368 TI - Central nervous system (CNS) metastasis from ovarian carcinoma. PMID- 9345369 TI - Donald H Silberberg, MD. PMID- 9345370 TI - In vitro studies of glial cells: what can we learn about demyelinating diseases? AB - Acquired demyelinating diseases of the central and peripheral nervous systems comprise an important group of neurologic diseases of unknown etiology and incompletely understood pathogenesis. Cultures of glial cells are proving highly useful in investigating the role of both antibodies and cytokines in the pathogenesis of these disorders. While there clearly is need for comparative studies employing more complex systems and using patient derived tissues, glial cell cultures provide important advantages by allowing researchers to characterize the effect of cytokines and growth factors on specific cell types in controlled conditions. PMID- 9345371 TI - The search for virus in multiple sclerosis brain. AB - Plaque-periplaque areas from MS brain tissue were explanted and propagated in tissue culture. The same in vitro techniques that successfully rescued measles virus from SSPE brain, papovavirus from PML brain, and HSV from normal human trigeminal ganglia, were applied. MS brain cells were also inoculated into chimpanzees, multiple rodent species, and embryonated hens eggs. No neurologic disease developed in experimentally infected animals, and no cytopathic effect was observed in explanted cells, or after cocultivation or fusion of MS brain cells with indicator cells. Further analysis of explanted and cocultivated cells by indirect immunofluorescence with various antiviral antisera prepared against viruses associated with post-infectious encephalomyelitis, as well as antisera to other ubiquitous viruses, failed to detect viral antigen. Finally, attempts to detect a latent enveloped virus in MS brain cells by 'superinfecting' MS brain cells in culture with vesicular stomatitis virus (VSV) did not reveal a VSV non neutralizable fraction. Nevertheless, since oligoclonal bands (OGBs) in the CSF of patients with chronic infectious diseases of the CNS are directed against the causative agent, it is likely that OGBs in MS CSF are antibody directed against the agent or antigen that triggered disease. Although the relevant antibody may be scarce relative to irrelevant antibody in MS CSF, and only small amounts of an MS-specific antigen may be present in brain, this report provides a rationale for strategies proposed in our companion report by Owens et al which will allow detection of an MS-specific antigen or its cognate RNA in brain. PMID- 9345372 TI - Strategies to identify sequences or antigens unique to multiple sclerosis. AB - Chronic inflammatory and infectious diseases of the central nervous system (CNS) are characterized by increased IgG and oligoclonal bands (OGBs) in the brain and cerebrospinal fluid (CSF). The OGBs in CNS infectious diseases of known cause have been shown to be directed against the pathogenic agent. In multiple sclerosis (MS), the antigenic specificity of the OGBs is unknown, but could be directed against an infectious agent, an autoantigen, or both. In a molecular approach to identify antigens specific for MS, we constructed directional cDNA expression libraries with mRNA extracted from chronic and acute MS plaques and periplaque white matter. The libraries were: (1) screened to identify clones whose expression products react with MS CSF, but not with CSF from other infectious and inflammatory diseases of the CNS; (2) subtracted by hybridization to mRNA from normal human brain white matter and differentially screened to detect unique MS transcripts; and (3) used as template in polymerase chain reactions to amplify, clone, and sequence IgG heavy and light chain variable regions (VH and VL, respectively) expressed in MS plaques. Analysis of the VH and VL IgG repertoire in MS brain may identify disease-relevant IgG sequences that can be assembled into functional antibodies using recombinant phage technology. Such recombinant antibodies will be useful to probe brain tissue to identify antigens unique to MS. PMID- 9345373 TI - Multiple sclerosis: prospects for remyelination. PMID- 9345374 TI - MRI studies of multiple sclerosis: implications for the natural history of the disease and for monitoring effectiveness of experimental therapies. AB - The use of magnetic resonance imaging (MRI) in multiple sclerosis (MS) has increased in our understanding of the natural history of the disease course and has provided and important tool for the analysis of new experimental therapies. Studies using MRI as well as pathological studies of MS indicate that the first event in the development of a new MS lesion as seen on T2 weighted images is disruption of the blood brain barrier (BBBD) which can be demonstrated by areas of increased signal on T1 weighted images done after the administration of gadolinium DTPA. When GdDTPA enhanced MRIs are used to monitor disease activity in patients with mild relapsing remitting MS, a considerable degree of disease activity is observed in clinically stable patients. These findings indicate that MS is an active and progressive disease in most patients even during the earliest phases of the disease and before significant clinical disability has occurred. MRI is also an important tool in evaluating new therapies. Using simple baseline vs treatment designs evidence for an effect of a new treatment on MRI parameters such as Gd-DTPA enhanced measure of BBBD can be achieved using a small study cohort and over a short duration. Together these advances should lead to more rapid progress in the understanding of MS and in identifying new treatments. PMID- 9345375 TI - Immunomodulation with linomide: possible novel therapy for multiple sclerosis. AB - Linomide, a synthetic quinoline carboxamide, has the ability to stimulate various lymphocyte subpopulations. We have shown its inhibitory effect on the clinical and histological signs of acute and chronic relapsing EAE. In these models linomide induces suppression of lymphocyte response to antigens, production of autoantibody, antigen presentation to specific T-cell lines and Mac-I expression, and induces activation of NK and suppressor-inducer cells. We have subsequently shown its inhibitory effect on clinical and MRI signs of patients with secondary progressive multiple sclerosis. Results of a double blind, placebo controlled, short term pilot study with p.o. linomide, showed a significant effect on the clinical disability scale (EDSS) (P = 0.045) and on the mean total number of new lesions in serial monthly MRI scans (P = 0.021). The increase of CD45Ra, CD8 and CD16 positive cells in linomide treated patients may indicate the importance of suppressor-inducer, suppressor and NK cells for the inhibition of the autoimmune response in the disease. PMID- 9345376 TI - Transcription of myelin basic protein promoted by regulatory elements in the proximal 5' sequence requires myelinogenesis. AB - Myelination in the central nervous system requires synthesis by oligodendrocytes of enormous amounts of lipids and proteins for incorporation in the developing myelin membranes. To approach the regulatory events coordinating the transcriptional activation of the genes that encode myelin proteins, we examined control of the myelin basic protein (MBP) locus. MBP plays a major role in myelin compaction. During development, MBP is already expressed in mature non myelinating oligodendrocytes. Here we show that, in transgenic animals in which the E. coli lacZ reporter gene is under the control of increasingly large portions (256, 1900 and 3200 bp) of the MBP promoter, 5' of the initiation of transcription site, reporter gene expression was initiated after myelin formation had started. This delayed expression of the transgene compared to MBP, strongly suggests that premyelinating expression is dependent on regulatory elements located outside of the 3200 bp sequence studied, while expression occurring at the time of myelin formation is dependent on the proximal promoter sequence. PMID- 9345377 TI - Possible association between multiple sclerosis and the human T cell leukemia virus (HTLV)-related endogenous element, HRES-1. AB - In the present study we searched for an association between the human endogenous retroviral element HRES-1 and multiple sclerosis (MS). Fragments of this endogenous retrovirus were amplified for subsequent examination by single strand conformational analysis. We did not find HRES-1 markers exclusively linked with MS and only the two already known polymorphisms, which define three alleles of HRES-1, were detected. However, we found a significant difference in the distribution of these alleles between a group of 87 MS patients and a control group of 158 healthy individuals (P = 0.014). There were no differences in the distribution of the HRES-1 allelic forms between MS patients with a relapsing remitting course and patients with chronic progressive MS. Our results provide evidence of an association between HRES-1 and MS. Possible explanations for this are discussed. PMID- 9345378 TI - Primary progressive multiple sclerosis: a distinct syndrome? AB - Multiple sclerosis (MS) has long been recognised to have both a relapsing remitting and progressive course. More recently patients with progressive disease have been further sub-divided into those with a progressive course from onset (primary progressive MS) and those with progressive decline following an initially relapsing-remitting period (secondary progressive MS). Diversity in MS may not however be restricted to clinical course. There is growing evidence that the subgroups of MS also differ with respect to clinical features, epidemiology, pathogenesis, genetics and neuroimaging appearances. In this review we outline the criteria variously applied in the classification of MS patients, addressing the need for a clear nomenclature. We evaluate the proposition that primary progressive MS has a profile distinct from other MS categories, contrasting the separate differential diagnoses and examining the implications for future therapeutic trials. PMID- 9345379 TI - Comparative evaluations of neuroperformance and clinical outcome assessments in chronic progressive multiple sclerosis: I. Reliability, validity and sensitivity to disease progression. Multiple Sclerosis Study Group. AB - There remains controversy regarding the most sensitive and valid outcome assessments to use in multiple sclerosis (MS) clinical trials. A double blind, placebo controlled, parallel group multicenter clinical trial to evaluate the clinical efficacy of cyclosporine A in chronic progressive MS incorporated several major clinical and performance outcome assessment modalities and a large sample size, both of which provide a unique opportunity to explore the relationship among MS disease status and the various outcome measures over time. The measures included a structured neurological examination, the Kurtzke Functional System scales and Expanded Disability Status Score, and the Incapacity Status Scale from the MS Minimal Record of Disability, the Harvard Ambulation Index, and neuroperformance testing. A test-retest reliability index, principal component analyses and a signal-to-noise ratio metric were used to comparatively evaluate the reliability, validity and sensitivity to disease progression of the various outcome assessments. The goal was to provide a rational basis for selection of behavioral outcome assessments in future MS clinical trials by identifying the primary dimensions of MS measured by the candidate outcome assessments and providing an objective basis for selecting tests that are most sensitive to MS disease and its progression over a two year trial period. We conclude that the components of the major clinical and performance measures show excellent reliability and cross validation. Principal component analyses of all outcome assessments yielded six primary underlying factors for describing disease status in chronic progressive MS that included lower extremity/pyramidal dysfunction, cerebellar/brainstem and upper extremity dysfunction, somatosensory dysfunction, visual dysfunction, mental or intellectual dysfunction and bowel/bladder problems. Signal-to-noise ratios indicated that upper and lower extremity composites of neuroperformance test items provided the most sensitive indicators of MS disease progression in the placebo group over the 2 year trial period. PMID- 9345380 TI - Desferrioxamine in chronic progressive multiple sclerosis: a pilot study. AB - Chronic progressive Multiple Sclerosis is refractory to many conventional treatments. We performed a pilot study testing desferroxamine (DFO) as a candidate in the treatment of chronic progressive Multiple Sclerosis. DFO was given daily by 8 h subcutaneous infusions at a dose of 2 grams daily for 7 days, followed by 1 gram daily for 7 days. Eighteen of 19 individuals completed the full dose of 21 grams. One patient was unable to complete the course due to nausea. No acute deterioration of neurological status was seen during the administration of DFO. No worsening of vision or hearing was noted except that the one patient who was unable to tolerate the medication had a transient reduction in hearing. All patients had a local redness at the injection site. None of the patients had any sudden worsening during or shortly after the treatment. This pilot study suggests that DFO is relatively well tolerated by Multiple Sclerosis patients when given in a short course of therapy. PMID- 9345381 TI - Energy cost of exercise in multiple sclerosis patients with low degree of disability. AB - In 10 patients (five females) suffering from multiple sclerosis with mild degree of disability, (EDSS ranging from 0 to 2) and in 10 age and sex matched control subjects we investigated lung function, respiratory muscles strength and cardiorespiratory response to incremental exercise in order to assess the metabolic cost of exercise. In the absence of any impairment of lung volumes and flows and in- and expiratory maximal mouth pressures, at peak of exercise oxygen consumption (VO2max = 1886 +/- 145 ml/min) and workload (Wmax = 137 +/- 9.8 watts) were slightly diminished in patients, as compared with controls (VO2max = 2246 +/- 196 ml/min and Wmax = 164 +/- 14.7 watts). These findings were associated with an increased heart rate (HR) and reduced oxygen pulse (VO2/HR) at the same workloads. During the whole exercise, however, the slope of the linear relationship between VO2 and work exhibited by the patients, amounting to 9.9 +/- 0.6 ml/min/watt, was similar to that of the controls (10.9 +/- 0.42 ml/min/watt). Incidentally, both at rest and during exercise, the patients showed a significantly greater minute ventilation (VE) due to a faster respiratory rate, associated with an augmented dead space (P < 0.05). We conclude that an increase of metabolic cost of exercise does not occur in multiple sclerosis patients with mild disability, suggesting a lack or a low degree of spasticity and/or ataxia elicited by the effort. Thus, their exertional capacity appears to be limited mainly by a poor training. The tachypnea observed in these patients at rest and during exercise was unexpected and the reason for adopting such a pattern of breathing is unclear. PMID- 9345382 TI - Neuropsychological assessment in multiple sclerosis: methodological issues and concerns. AB - A brief overview of research findings in five key areas pertaining to the study of neuropsychological aspects of multiple sclerosis is presented. The areas covered are: (1) general and specific cognitive deficits (2) prevalence of these deficits (3) attempts to correlate these deficits with clinicopathological features (4) attempts to investigate the relationship between deficits and abnormalities detected on magnetic resonance scanning and (5) longitudinal studies of cognitive deficits and MS. Based on this review, methodological issues that continue to hinder comparison between research reports, and across research centres, are outlined. Problem areas identified are (1) patient selection (2) selection of control subjects (3) lack of consensus regarding test selection (4) problems with statistical procedures (5) the relationship between depression and cognition (6) problems with measuring disability and MR abnormalities and (7) lack of longitudinal data. PMID- 9345383 TI - Investigational drug therapies of treatment of multiple sclerosis. AB - The licensing of interferon beta-1b dramatically changed the treatment of multiple sclerosis (MS) in the United States. Although it was the first therapeutic agent shown to affect the natural course of the disease, interferon beta-1b is not appropriate for all patients and is far from being a cure. Several other promising therapies now under study include immunosuppressive and immunomodulatory drugs to limit inflammation; oral administration of myelin to induce tolerance; monoclonal antibodies designed to deliver targeted immunotherapy; potassium channel blockers to facilitate conduction along demyelinated axons; and glial growth factors to promote remyelination. Clinical trials of potential therapeutic agents have proliferated in the past decade in conjunction with rapid advances in our understanding of the immunologic basis of MS. Some investigational therapies are associated with problematic toxicities, others benefit only a minority of patients, and many are still in the early stages of development. Nevertheless, because current therapeutic options are limited, and because the history of MS therapy is one of disappointment and frustration, it is essential that legitimate, scientifically based advances be widely disseminated to the neurologic community. This article reviews some of the most promising current and investigational therapies for MS. PMID- 9345384 TI - Oligoclonal expansion of gamma delta T cells in cerebrospinal fluid of multiple sclerosis patients. AB - We have used a PCR based method to analyse TCR gamma chain repertoire and clonality of gamma delta T cells in the CSF and blood of II MS patients. Samples collected from nine patients with other neurological diseases were used as a control. Five controls had central nervous system inflammation and four had non inflammatory processes. We have observed a decreased percentage of gamma delta T cells expressing TCR gamma with V gamma 9 and J gamma P fragments in the CSF samples in comparison with the blood. We did not final clonal expansion of the gamma delta T cells in any control case. Clonal expansion of gamma delta T cells occurred in five of II MS cases in the CSF but not in the blood. Two of these clones expressed TCR gamma rearranged with V gamma 9 and J gamma 1 fragments, two others used V gamma 10 and J gamma P1, and one used V gamma 9 and J gamma P fragments. We found no correlation between clonality and clinical state of patients, duration of the disease or number of cells in CSF. Our study provides additional evidence for the possible role of the gamma delta T cells in the MS pathogenesis. PMID- 9345385 TI - Survey of herpes simplex virus infections of the central nervous system, including acute disseminated encephalomyelitis, in the Kyushu and Okinawa regions of Japan. AB - We analysed data from 27 patients with herpes simplex virus (HSV) infections of the central nervous system (CNS) found in a 1990-1992 survey in Kyushu and Okinawa, Japan. Patients ranged in age from one year to 70 years, with peaks seen in the 20s and 50s. Temporal lobe-limbic encephalitis was the most common HSV infection (13 patients), followed by meningitis (5), diffuse encephalitis (4), disseminated encephalomyelitis (ADEM) (3) and brain stem encephalitis (2). Another three patients with non-herpetic, non-paraneoplastic acute limbic encephalitis were presented. Our study indicates that HSV infection can course ADEM, although temporal lobe-limbic encephalitis or meningitis are more common. The early diagnosis of HSV-related ADEM is important because of the efficacy of the timely administration of corticosteroids. PMID- 9345386 TI - Multiple sclerosis and HTLV-I associated myelopathy/tropical spastic paraparesis are two distinct clinical entities. AB - Multiple sclerosis (MS) and HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) can overlap in their clinical features and thereby cause difficulties for clinicians in relation to diagnosis and therapy. However, epidemiological biochemical, immunological, virological and radiological studies point to a number of significant differences. Recent comparative neurophysiological data, including blink reflex studies, obtained in these disorders, is briefly reviewed here and provides additional evidence of difference. The abnormal blink reflex in patients with MS consist of prolonged latencies and absences of R1 and R2 responses and are mainly due to demyelinating lesions around the pans. In contrast, in HAM/TSP the blink reflex abnormalities frequently include an unusual early response, R1k, which is probably a consequence of interneuronal hyperexcitability around the brainstem. Thus these findings provide further support for our contention that HAM/TSP and multiple sclerosis are distinctly different both as clinical entities and in their underlying pathomechanisms. PMID- 9345387 TI - Long-term cultured astrocytes inhibit myelin formation, but not axonal growth in the co-cultured nerve tissue. AB - The chronic demyelinated plaque of multiple sclerosis (MS) is characterised by a loss of oligodendrocytes, astrogliosis, and incomplete or no remyelination which probably results in part from the suppressive effects of gliotic astrocytes on myelin formation. We explanted mouse cerebella on astrocyte cultures which had been maintained for 2 to 12 weeks and assessed the myelination in the cerebellar tissue at 18 days after explanation. Myelination occurred vigorously in the tissue explanted on 2- to 4-week-old astrocytes, but was poorer in the tissue explanted on astrocytes older than 4 weeks. No myelin sheath was formed on 12 week-old astrocytes, although axons developed equally as well as those in the tissues explanted on 2-week-old astrocytes. As astrocytes were maintained longer, they became fibrous and immunostained more deeply with anti-glial fibrillary acidic protein antibody, being analogous to astrogliosis. These findings imply that astrogliosis in chronic demyelinated lesions of MS may potentially block remyelination. PMID- 9345388 TI - Interferon-beta in the treatment of multiple sclerosis. Symposium proceedings. Brussels, Belgium, September 23-24, 1994. PMID- 9345389 TI - Interferon beta in the cytokine network: an anti-inflammatory pathway. AB - Interferons are cytokines and thus fulfil a vital role in communication between cells in their microenvironment. Type I interferons, the group to which interferon beta (IFN-beta) belongs, share several structural and functional properties by which they distinguish themselves from type II interferon or IFN gamma. In particular, IFN-beta can be produced by many different cells while IFN gamma is an exclusively lymphocytic cytokine, i.e. a lymphokine. IFN-beta is functionally linked to other cytokines as it can induced by some of them (e.g. interleukin I) and as its actions can be potentiated or antagonized by other cytokines. Such interactions can take place at several levels, e.g. at the level of signal transduction and transcription activation. Of potential interest for the role of IFN-beta in multiple sclerosis is its ability to function as a deactivator of mononuclear phagocytes, and hence as an inhibitor of inflammation. PMID- 9345390 TI - Antagonism of interferon beta on interferon gamma: inhibition of signal transduction in vitro and reduction of serum levels in multiple sclerosis patients. AB - Interferon gamma (IFN-gamma acts as a mediator of multiple sclerosis (MS) exacerbations through a number of biological effects, such as induction of major histocompatibility class II complexes (MHC-II), macrophage activation and potentiation of tumor necrosis factor (TNF-alpha). The clinical efficacy of interferon beta (IFN-beta) therapy in reducing exacerbations of relapsing remitting MS has been related to antagonistic effects on various activities of IFN-gamma, including MHC-II gene induction. However, there is no model to explain such antagonistic effects of IFN-beta and IFN-gamma, and the two cytokines are also known to act synergistically against viruses and in the induction of MHC-I. We show that IFN-beta does inhibit an immediate molecular event of IFN-gamma, namely activation and DNA binding of the transcription factor Stat1. We propose a model of direct interference of the IFN-gamma and IFN-alpha,beta signal transduction pathways accounting for antagonistic effects on some genes, which in turn activate MHC-II transcription, as well as for synergistic effects on other genes. In addition, study of MS patients treated with natural IFN-beta shows that IFN-beta significantly reduces serum levels of IFN-gamma while increasing IL-4, strongly suggesting that IFN-beta also controls the relative activation of TH1- and TH2-type T lymphocytes. PMID- 9345391 TI - Biologic effects of interferons: relevance to multiple sclerosis. AB - Recombinant interferon beta has established efficacy for relapsing-remitting MS, but the mechanisms of action in the disease are not well understood. Interferons (IFNs) mediate biologic effects by receptor-mediated gene activation. Binding by IFNs to high-affinity surface receptors results in physophorylation and activation of two cytoplasmic tyrosine kinases. This leads to activation of latent transcription factors in cell cytoplasm that translocate to the nucleus, where activated transcriptional elements interact with the interferon-stimulated response element (ISRE), leading to transcription of the interferon-stimulated genes (ISGs). IFN's biologic effects are mediated by function of the ISGs. The biologic effects of IFNs have been classified as antiviral, antiproliferative and immunomodulatory; effects are complex, however, because there are two separate types of IFN, four separate varieties of type I IFN and approximately 30 known interferon-regulated genes. Known effects of IFN that may plausibly relate to therapy efficacy in MS are discussed. PMID- 9345392 TI - Early treatment trials with interferon beta in multiple sclerosis. AB - Recent reports of the successful treatment of relapsing-remitting MS with interferon beta-1b (IFN-beta) have ushered in a new era of immunotherapy. In a sense, this was the result of a remarkable conjunction of molecular biotechnology, immunology and clinical research, resulting in the first therapeutic agent to alter the course of this previously untreatable disease. In more concrete terms, the use of IFN-beta in MS was the logical outcome of a series of clinical trials of natural and recombinant IFNs carried out over the past decade, and of concurrent laboratory research suggesting that the effects of the IFNs in MS are mediated by immunoregulatory rather than antiviral or antiproliferative mechanisms. It is now known that the proinflammatory cytokines IFN-gamma and tumor necrosis factor alpha (TNF-alpha) are probably involved in the pathogenesis of MS lesions. In contrast, type I IFNs (alpha and beta) tend to inhibit the activity of IFN-gamma and to prevent disease activity. The earliest controlled studies of natural IFN-alpha and IFN-beta, carried out in the early 1980s, led to the phase III clinical trial of systemic recombinant IFN-beta (Betaseron), recently completed in the United States and Canada. In patients treated with high-dose IFN-beta there were significant reductions in relapse rate and in the appearance of new lesions on magnetic resonance imaging (MRI). The US Food and Drug Administration approved Betaseron for treatment of relapsing remitting MS in 1993, and it is now in widespread clinical use. A trial of another recombinant IFN-beta, given by intramuscular injection once a week, was also recently completed. The results of this study are awaited with great interest because the primary end point was progression of disability rather than relapse rate. Meanwhile, recombinant IFN-alpha was reported to prevent relapses and MRI changes in a small but well-designed trial. In this paper, the early clinical studies and some of the immunological developments leading to the use of IFN-beta in MS are reviewed. PMID- 9345393 TI - Magnetic resonance imaging as an outcome measure in the treatment of multiple sclerosis: results of the interferon beta trial. PMID- 9345394 TI - Early treatment of multiple sclerosis with Rebif (recombinant human interferon beta): design of the study. AB - Two recent properly controlled trials performed in patients with relapsing remitting MS have demonstrated that interferon beta substantially improves the evolution of the disease. The results of the Optic Neuritis Treatment Trials suggest that early treatment of patients with isolated optic neuritis may delay the conversion to clinically definite MS (CDMS). Moreover, clinical trials in MS and in immune-mediated diseases indicate that better results are obtained in patients in the early phases of the disease. We present the design of a multicentre, randomized, double-blind, placebo-controlled study of recombinant interferon beta (Rebif-Serono) in patients with a first attack suggestive of MS. The primary objective of the study is to investigate the efficacy of Rebif, administered subcutaneously at the dose of 8 million international units (MIU) once a week for 2 years, on the risk of developing CDMS. PMID- 9345395 TI - Human recombinant interferon beta in the treatment of relapsing-remitting multiple sclerosis: preliminary observations. AB - An open comparative, randomized trial with recombinant human interferon beta (r hIFN-beta) involving 72 patients with clinically definite and/or laboratory supported relapsing-remitting MS is in progress at the University 'La Sapienza' and at the S. Camillo Hospital of Rome. After a 6 month period of clinical and magnetic resonance imaging (MRI) observation (baseline findings), patients are randomly assigned to one of two treatment groups receiving 3 or 9 MIU of recombinant human IFN-beta (r-hIFN-beta) self-administered by subcutaneous injection three times a week for 6 months. All patients are examined by MRI with and without gadolinium (Gd-DTPA) every 4 weeks for the entire duration of the study (12 months). The main aim of the study is to test the hypothesis that r hIFN-beta may halt or slow the progression of the disease by showing a significant reduction in MRI activity. This will be achieved by comparing pre and post-treatment periods. As an additional clinical end point, the exacerbation rate during these two periods will be compared. This study began in June 1993 and the final analysis of MRI data is planned for the spring of 1995. PMID- 9345396 TI - Interferon alpha treatment of relapsing-remitting multiple sclerosis: long-term study of the correlations between clinical and magnetic resonance imaging results and effects on the immune function. AB - Interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) are proinflammatory cytokines which may be involved in the pathogenesis of MS. IFN alpha counteracts many of the proinflammatory actions of IFN-gamma and TNF-alpha. We treated 20 patients with relapsing-remitting (RR) MS with 9 MIU of recombinant IFN-alpha-2a (rIFN-alpha) (n = 12) or placebo (n = 8) intramuscularly every other day for 6 months. Clinical exacerbations or new or enlarging lesions at serial MRI occurred in 2/12 rIFN-alpha-treated and in 7/8 placebo-treated patients (P < 0.005). Only one new MRI lesion was detected in the rIFN-alpha group, while 27 new or enlarging lesions were detected in placebo group (P < 0.01). Baseline lymphocyte IFN-gamma (19.10 +/- 7.12 U ml-1) and TNF-alpha (18.05 +/- 5.34 pg ml 1) production significantly decreased to 3.03 +/- 0.66 (P < 0.04) (for IFN-gamma) and to 5.78 +/- 0.90 (P < 0.04) (for TNF-alpha) after rIFN-alpha treatment. IFN gamma and TNF-alpha production was unchanged in the placebo group. rIFN-alpha was tolerated without drop-outs or serious side-effects, but fever, malaise, fatigue (interfering with daily activities in two patients) and leukopenia frequently occurred. High-dose chronic systemic rIFN-alpha might reduce clinical and MRI signs of disease activity in RRMS. The changes in cytokine production suggest that the effect is probably mediated by a down-regulation of proinflammatory cytokine. PMID- 9345398 TI - Human interferon omega--a review. AB - A single functional gene in the human genome codes for interferon omega (IFN omega), a monomeric glycoprotein distantly related in structure to IFN-alpha and IFN-beta, but unrelated to IFN-gamma. IFN-omega is secreted by virus-infected leukocytes as a major component of human leukocyte interferon. The human class I IFN receptor complex which mediates the biological activity of IFN-alpha and IFN beta also binds IFN-omega. Its specific activity in a standard in vitro antiviral assay system is 4 x 10(8) U mg-1; potent antiviral activity against several DNA and RNA viruses has been demonstrated. IFN-omega inhibits proliferation of a variety of tumor cell lines in vitro. The protein stimulates natural killer cell activity, enhances expression of major histocompatibility complex class I (but not class II) antigens and inhibits proliferation of lymphocytes stimulated with mitogens or allogeneic cells. IFN-omega is unrelated to IFN-alpha, -beta and gamma in its antigenic properties, as it does not cross-react with antisera or monoclonal antibodies in immunoassays or antiviral neutralization bioassays. Antibodies induced in patients by long-term IFN-alpha 2 therapy that block IFN alpha 2 activity do not inactivate IFN-omega. As IFN-omega, like other human IFNs, has a species-restricted biological activity, evaluation of its therapeutic potential will have to await clinical trials. PMID- 9345399 TI - Recombinant human interferon beta in the treatment of relapsing-remitting and secondary progressive multiple sclerosis. AB - Two multicenter, randomized, double-blind, placebo-controlled phase III studies to evaluate the safety and efficacy of 6 and 12 MIU of recombinant human interferon beta (r-hIFN-beta) (Rebif) given subcutaneously three times per week, in patients with MS, are described. In one study, patients with relapsing remitting multiple sclerosis are treated for 24 months; the primary efficacy end point is the number of exacerbations. In the other study patients with secondary progressive multiple sclerosis are treated for 36 months; the primary efficacy end point is time to sustained progression in disability as determined by the Expanded Disability Status Scale (EDSS). Both studies are conducted in centers in Europe, Australia and Canada. PMID- 9345397 TI - Effect of interferon gamma on T lymphocytes from patients with multiple sclerosis. AB - A central role in the complex immunology of MS is played by activated T lymphocytes. Proper antigenic stimulation, adequate major histocompatibility complex (MHC) coactivation and multiple cytokine signals regulate T-cell activation via the generation of intracellular calcium (Ca2+) transients necessary to up-regulate the genes controlling lymphocyte growth and differentiation. Interferon gamma (IFN-gamma) exerts an autocrine/paracrine control of T-lymphocyte activity and is able to co-mediate most demyelinating events occurring in MS patients. The mechanisms by which IFN-gamma exerts its effects include increased expression of MHC class II molecules on the surface of glial cells and stimulation of macrophage/microglial cell production of molecules toxic to myelin. Most intracellular events regulating these processes in lymphocytes are, however, still unknown. We have reported that in the majority of MS patients T lymphocytes (mainly CD4+) exposed to IFN-gamma show a Ca2+ influx whose ion permeability and pharmacological properties differ from those of all Ca2+ influxes so far described. This IFN-gamma-activated Ca2+ influx, which is probably sustained by a cationic channel, was observed in the majority of patients with MS, but only in a limited number of subjects affected by other neurological, active immune-mediated diseases or healthy control subjects. Moreover, the presence of the influx correlates with clinical and radiological evidence of disease activity and induces T-lymphocyte proliferation even when cells are suboptimally stimulated by activatory stimuli. We conclude that this new IFN-gamma-activated Ca2+ influx seems to be highly specific for MS (especially during its active phase), and could be important for the intracellular regulation of cells involved in demyelination. The presence of the influx in T lymphocytes from MS patients could account for the reported temporal association between infections and clinical relapses. PMID- 9345401 TI - Guidelines for early treatment trials in patients presenting with clinical and paraclinical abnormalities which put them at high risk for conversion to multiple sclerosis. European Charcot Foundation Working Group for Treatment Trials. AB - Several recent clinical trials have shown that interferon beta, a rather well tolerated therapy, can favourably alter the pathological disease activity in MS, decrease the frequency of relapses and slow down the progression of the disability. Clinical and biological observations suggest that immune abnormalities in MS can be corrected more easily during the first stages of the disease, and thus early treatments may be more effective. In addition, the results of the Optic Neuritis Treatment Trial suggest that administration of a therapy immediately after the first attack may delay the occurrence of subsequent signs and/or symptoms and, by the same token, the conversion to clinically definite MS. Moreover, we know that the clinical activity of the disease during the early stage is strongly predictive of the subsequent evolution. For all these reasons, it seems thus justified to evaluate whether treatment with effective and well-tolerated drugs can retard the long-term disability and improve the quality of life of patients who present with clinical signs and paraclinical tests which put them at high risk of developing MS. PMID- 9345400 TI - Interferon beta-1b in secondary progressive multiple sclerosis--outline of the clinical trial. AB - This manuscript describes the outline of a double-blind, placebo-controlled, (European), multicentre phase III study to evaluate the safety and efficacy of 8 MIU of interferon beta-Ib given subcutaneously every other day for 3 years in patients with secondary progressive multiple sclerosis. The primary efficacy variable of this trial is the time to confirmed neurological deterioration as documented by the Expanded Disability Status Scale. The essentials of the study design are presented, including the rationale for the performance of the study and the selection of both clinical and magnetic resonance imaging outcome parameters. PMID- 9345402 TI - Data handling in clinical trials: an ongoing debate. AB - Data handling is a crucial phase in controlled clinical trials. In this paper we conceive the process of data handling in a broad sense, encompassing monitoring, analysis and publication of data. We discuss basic ethical principles underlying adequate data handling and suggest some modifications of the Good Clinical Practice guidelines. As to data monitoring, we underline the corporate responsibilities and argue in favour of an independent Data and Safety Monitoring Board (DSMB). Data analysis should be performed in independent institutions, and there is a moral obligation to perform an interim analysis. Regarding accessibility of data, the raw data should be made available to participating researchers and independent scientific institutions, and both positive and negative results should be published, in agreement with a Report on Drug Information by the World Health Organization. To meet these requirements, commercial sponsors should have initiatives for independent DSMBs, interim analysis and complete data availability, and conceive corresponding contracts. The investigators can improve the quality of data handling by forming an Investigators Liaison Panel that represents them in the contacts with the sponsor. Finally, ethics committee and drug regulatory agencies should see to it that newly gained insights are being put into practice. PMID- 9345403 TI - Interferon beta in the treatment of multiple sclerosis--closing remarks. PMID- 9345404 TI - Treatment of relapsing-remitting multiple sclerosis with natural interferon beta: a multicenter, randomized clinical trial. AB - Preliminary results of a multicenter, randomized study on the effect of natural interferon beta (n-INFN-beta) in relapsing-remitting MS are reported. Sixty patients received either no treatment or n-IFN-beta (9 MIU three times a week, subcutaneously). After 6 months, the n-IFN-beta-treated group showed a 58% median reduction in the number of active lesions detected on monthly magnetic resonance imaging scans as well as a 38% reduction in the exacerbation rate as compared with the control group. Side-effects were mild and self-limiting. The study continues according to the protocol. PMID- 9345405 TI - Introductory remarks: immunosuppressive and immunomodulating drugs, where and how do they act? AB - Immunotherapies used in multiple sclerosis are reviewed. The mechanisms of action supporting their clinical application are described according to our current understanding. Immune treatments can be divided into three groups: (1) cytostatic and cytotoxic immunosuppressants which block the cell cycle of immunocompetent cells and/or provoke their deletion; (2) immunomodulators which specifically interfere in cellular or humoral immune mechanisms; (3) immunoregulators which restore immunodeficient states but have also immunosuppressive properties at the same time. Numerous attempts have been made to correct almost all abnormal immune mechanisms underlying disease progression. The results of those clinical trials are reviewed. Recent studies have demonstrated that a severe, unspecific and sustained immunosuppression markedly downregulates the clinical and pathological activity of the disease. Unfortunately, serious delayed adverse effects prevent a long-term administration. Recent specific immunomodulators with an acceptable toxicity have provided modest but unquestionable benefit on the attack rate and MRI lesion burden. No clear effect on progression has been demonstrated to date. There is still much work to be done to improve the efficacy of our current therapies on the attack rate and particularly on the progression. Several promising new compounds are already under evaluation. Another approach is combination therapies which will certainly become critical in MS like in other autoimmune diseases. PMID- 9345406 TI - Interferon beta-1b: latest published results, 1995. AB - Interferon beta-Ib (Betaseron) has been shown in a randomized, double-blind, placebo-controlled study to reduce relapse rate, relapse severity and MRI progression in patients with relapsing-remitting MS. The recently published Betaseron extension trial and studies from the National Institutes of Health provide additional evidence suggesting an important treatment effect. The major new findings from these studies are reviewed in this paper. Other findings from these recent studies merit attention, however. Specifically, neutralizing antibody formation was seen in 38% of patients receiving high-dose Betaseron at 3 years. With the development of these antibodies, there was no longer clinical evidence that the drug remained effective. This observation must be considered carefully when initiating Betaseron therapy. The Betaseron trial suggests that MRI is an imperfect predictor of clinical disease activity. Of particular interest was the finding that patients receiving low-dose Betaseron developed 'confirmed treatment failure' sooner than placebo-treated patients, despite a clear treatment benefit on MRI-detected evidence of subclinical disease activity. PMID- 9345407 TI - Interferon beta treatment in multiple sclerosis: the European clinical trials. PMID- 9345409 TI - Management of relapsing/remitting multiple sclerosis with copolymer 1 (Copaxone). AB - Copolymer I (Copazone) was evaluated in a multicenter, placebo-controlled, double blind trial at 11 universities. Two hundred and fifty-one relapsing-remitting ambulatory MS patients were randomized to receive 20 mg of copolymer I or placebo by daily subcutaneous injection for approximately 30 months. At conclusion, the copolymer I group had 32% fewer relapses (P = 0.002) and significantly more were relapse-free (P = 0.035). Significantly, more patients were receiving copolymer I had improved during the study, while more patients on placebo showed neurological decline (P = 0.001). There were few side effects and no drug related laboratory abnormalities. Copolymer I is being considered by North American and European regulatory agencies for approval as commercially available agent for the control of multiple sclerosis. PMID- 9345408 TI - Interferon beta treatment for multiple sclerosis: persisting questions. AB - Doctors Noseworthy and Summerfield have reviewed the results of placebo controlled, double-masked, clinical trials of interferon beta-1b (IFNB-1b, Betaseron) and interferons beta-1a (IFNB-1a, Avonex,) in patients with relapsing multiple sclerosis (MS). IFNB-1b, administered subcutaneously every other day to ambulatory patients with relapsing-remitting MS, significantly reduces annual exacerbation rate and percentage change from baseline MRI T2-weighted total lesion area. There is also evidence that IFNB-1b reduces the number of new gadolinium-enhancing lesions in patients with relapsing-remitting MS. IFNB-1a (Avonex), administered intramuscularly every week to patients with relapsing MS, significantly reduces annual exacerbation rate, the number and volume of new focal gadolinium-enhanced T1-weighted lesions, and slows the accumulation of physical disability over time. However, questions remain regarding the efficacy of these treatments, the significance of neutralizing antibody formation, indications for therapy, adverse events, optimal dose, and route of administration. Design limitations of the Phase III trial of IFNB-1b have been reviewed elsewhere, and a critical review of the results of the Phase III trial of IFNB-1a should be undertaken after the publication of the clinical and imaging results of that study. PMID- 9345410 TI - Methylprednisolone treatment in multiple sclerosis: effect of treatment, pharmacokinetics, future. AB - High dose intravenous methylprednisolone treatment is effective and safe in the treatment of relapses in multiple sclerosis, but the long term effects are unclear. Pharmacokinetics are almost unknown, but may be very important for the understanding of the clinical and paraclinical effects. In view of what is known now, IVMP should have a prominent place in basic and clinical MS research. PMID- 9345411 TI - Mitoxantrone immunotherapy in multiple sclerosis. AB - Mitoxantrone, a cytotoxic agent recently developed, was subsequently found a very potent immunosuppressor. Experimental data in experimental allergic encephaloymyelitis demonstrated a dramatic suppression of both active and passive forms. Immune effects concern cellular and humoral components and are particularly persistent. B cell subset is preferentially deleted. Suppressor cells are relatively spared and suppression becomes dominant. In cancer therapy, the main advantages of mitoxantrone are a definitely better immediate tolerance and very low delayed adverse reactions (carcinogenicity, teratogenicity, impact on reproductive organs). Given its major immunosuppressive activity and its better tolerance, mitoxantrone was a potential candidate for multiple sclerosis therapy. Several clinical trials have confirmed the remarkable efficacy of mitoxantrone to reduce both attack and progression rates. Unfortunately the cardiotoxicity was found more frequent than expected and limits the maximum cumulative dose to 120 mg/m2. Mitoxantrone, when employed properly, may be useful in patients with frequent and disabling excerbations and/or rapidly progressing disability. It must be kept in mind that multiple sclerosis is a chronic disease, and that the benefit is limited to the period of administration of any treatment. PMID- 9345412 TI - Guidelines for MRI monitoring of the treatment of multiple sclerosis: recommendations of the US Multiple Sclerosis Society's task force. AB - In relapsing-remitting and secondary progressive multiple sclerosis (MS), MRI activity on monthly brain scans is a sensitive primary outcome measure in short term exploratory treatment trials. Because conventional MRI findings have a limited correlation with disability, the primary outcome in definitive trials should be clinical, although MRI is useful in providing an index of pathological progression. In trials aimed at preventing evolution from a clinically isolated syndrome to MS, MRI findings should be used in the entry criteria. The likely pathological substrates of irreversible disability are demyelination and axonal loss. Putative MR markers for these pathologies appear to relate more closely to disability than conventional MRI findings. Further technical developments should lead to improved quantitation, pathological specificity and clinical correlations. PMID- 9345413 TI - A phase II trial of anti-CD4 antibodies in the treatment of multiple sclerosis. AB - In multiple sclerosis (MS) myelin damage is the result of a chronic inflammatory process mediated by CD4 positive T helper/effector cells. In experimental allergic encephalomyelitis (EAE), the animal model of MS, treatment with anti-CD4 antibodies can prevent the onset of disease. Natural history studies have demonstrated that gadolinium enhanced magnetic resonance imaging (MRI) of the brain is more sensitive and objective in assessing inflammatory disease activity in MS than clinical monitoring, so that less patients and shorter studies suffice to reach the same statistical power as compared to trials using clinical outcome parameters. In this paper we describe the design of an exploratory trial of chimeric monoclonal anti-CD4 antibodies in the treatment of MS. For this study we chose the number of active MS lesions on monthly gadolinium enhanced MRI scans as the primary outcome measure. PMID- 9345414 TI - Development of cladribine treatment in multiple sclerosis. AB - Cladribine is a new type of drug with properties of selective lymphocyte suppression that appear to favorably alter the clinical course of progressive multiple sclerosis (MS). The history of the development of cladribine treatment in chronic progressive MS is discussed, and the application of cladribine treatment to progressive multiple sclerosis in a double-blind, placebo crossover study is reviewed. Cladribine selectively targets both resting and dividing lymphocytes and may be able to destroy the activated lymphocytes that induce CNS demyelination, thus producing stabilization or improvement in chronic MS. Although the role of cladribine has not yet been fully defined, additional studies are underway to evaluate the efficacy and safety of cladribine in both progressive MS and relapsing-remitting MS. PMID- 9345415 TI - Linomide reduces the rate of active lesions in relapsing-remitting multiple sclerosis. PMID- 9345416 TI - Intravenous immunoglobulin G therapy: effects of acute and chronic treatment in multiple sclerosis. AB - High dose intravenous immunoglobulin (IVIG) exerts several effects on the immune system that could have a beneficial influence on the disease processes in multiple sclerosis (MS). IVIG may be useful in treatment of acute exacerbations, in prevention of new relapses, and in promotion of remyelination. Presently, the clinical evidence of effect of IVIG in MS is based on the results of small open trials, some of which, however, have been encouraging. Confirmation of a beneficial effect of IVIG must await the results of placebo-controlled, double blind trials currently ongoing in several centers. If effective, IVIG administration would be a valuable supplement to the existing treatment of MS. PMID- 9345417 TI - T cell vaccination in multiple sclerosis. AB - T cell responses to myelin basic protein (MBP) are implicated to play an important role in the pathogenesis of multiple sclerosis (MS). These MBP autoreactive T cells are found to undergo in vivo activation and clonal expansion in patients with MS. They accumulate in the brain compartment and may reside in the brain lesions of patients with MS. As MBP-reactive T cells potentially hold a central position in initiation and perpetuation of the brain inflammation, specific immune therapies designed to deplete them may improve the clinical course of the disease. In this paper, the therapeutic potential of T cell vaccination in the treatment of MS is discussed in context of its immunological and clinical effect. The results of our phase one clinical trial indicate that T cell vaccination with inactivated MBP autoreactive T cells induces specific regulatory T cell network of the host immune system to deplete circulating MBP reactive T cells in a clonotype-specific fashion. The immunity induced by T cell vaccination is clonotype-specific and long-lasting. Our longitudinal clinical evaluation further suggests a moderate reduction of rate of clinical exacerbation, disability score and the brain lesions (measured by magnetic resonance imaging) in vaccinated patients, as compared to matched controls. Our study should encourage further investigation on the treatment efficacy of T cell vaccination and further improvement for its clinical application. PMID- 9345418 TI - CAMPATH-IH in multiple sclerosis. AB - In a pilot study, seven patients with multiple sclerosis were treated with CAMPATH-IH which targets the CD52 antigen present on lymphocytes and monocytes. There was a substantial reduction in disease activity as measured by gadoliunium enhancing lesions on MRI. Encouraged by this result a further seven patients have been treated with CAMPATH-IH; four also received anti-CD4 antibody. Lymphopaenia developed rapidly and was sustained for at least one year. In 12 patients, the first infusion of antibody was characterised by significant exacerbation or re awakening of pre-existing symptoms lasting several hours. These clinical effects of antibody treatment correlated with increased levels of circulating cytokines. Peak levels of tumour necrosis factor alpha (TNF alpha) and interferon gamma (IFN gamma) occurred at 2 h whereas the rise in interleukin-6 (IL-6) was significantly delayed and peaked at 4 h after starting antibody treatment. The neurological symptoms could not be attributed directly to pyrexia and were not provoked (in one patient) by an artificial rise in temperature. In the remaining two patients, a single pre-treatment with intravenous methylprednisolone (500 mg) prevented both the transient increase in neurological symptoms and the cytokine release. Our results suggest that soluble immune mediators contribute to symptom production in multiple sclerosis by directly or indirectly blocking conduction through partially demyelinated pathways. PMID- 9345419 TI - Long term recombinant interferon alpha treatment in MS with special emphasis to side effects. AB - Twenty relapsing-remitting (RR) clinically definite MS patients were treated with 9 MIU intramuscular recombinant interferon alpha-2a (rIFNA) (Roferon-A, Roche) (n = 12) or placebo (n = 8) every other day for 6 months and followed up for a further 6 months after stopping treatment. Numbers of active lesions at MRI and of patients with clinical-MRI signs of disease activity and lymphocyte interferon gamma production, which were decreased during treatment, returned to values similar to baseline and placebos after stopping treatment. rIFNA chronic therapy seems therefore needed in order to maintain drug efficacy. Side effect profile was monitored, too, for over 1 year in the same 20 patients plus 25 additional RR MS patients. Besides the typical side effects of type I interferon therapy (fever, fatigue, depression, lymphopenia, hepatic enzyme elevation), occurrence of serum autoAbs was noted in 30% patients (in 60% antinuclear and in 80% antithyroid autoAbs). In two patients rIFNA treatment was stopped, in one case for antithyroid autoAbs and hypothyroidism, in the other for antinuclear autoAbs and a five-fold increase of ALT. A careful monitoring of serum autoAbs and of signs of thyroid or liver damage must always precede and accompany longterm type I IFN therapy. PMID- 9345420 TI - Treatment of relapsing-remittent multiple sclerosis with recombinant human interferon-alfa-2a: design of a randomised, placebo-controlled, double blind trial in Norway. AB - A multicentre, randomised, double-blind, placebo controlled study to evaluate the efficacy and safety of 4.5 and 9.0 MIU recombinant human interferon alfa-2a (Roferon-A) given thrice weekly in patients with relapsing-remittent multiple sclerosis is described. The patients are treated for 6 months followed by a 6 months drug-free period. The primary objective is to determine new disease activity analysed by monthly MRI with gadodiamide (GdDTPA-BMA, Omniscan). The study is conducted at eight centers in Norway and is completed in January 1996. PMID- 9345421 TI - Inhibitors of tumor necrosis factor-alpha: promising agents for the treatment of multiple sclerosis? AB - Tumor necrosis factor (TNF)-alpha is a critical inflammatory mediator of experimental autoimmune encephalomyelitis and multiple sclerosis, and may therefore be a useful target for immunotherapy. Therapeutic strategies aimed at TNF include pharmacological inhibitors of TNF synthesis and/or processing and biological inhibitors of TNF effects. Several anti-TNF agents are currently being tested in multiple sclerosis in pilot clinical trials. PMID- 9345422 TI - Emerging treatments in multiple sclerosis: azathioprine and mofetil. AB - Global immunosuppression instead of focused selective or specific immunomodulating strategies may still be relevant in diseases with chronic and broad immune dysregulation such as multiple sclerosis (MS). Among classical or new immunosuppressive drugs, two of them, both inhibiting purine synthesis, show an attractive profile for MS treatment. Azathioprine (AZA) is the most anciently and widely used global immunosuppressive drug in MS. Despite founded initial fears, it can be stated today that AZA is usually well tolerated and compatible with normal daily activities, that it requires minimal monitoring and does not significantly increase the risk of cancer induction after 5 years of continuous usage at the conventional 2.5 mg/kg daily dose. The only two presently available well conducted trials of AZA in ambulatory patients with relapsing-remitting MS show marginally significant beneficial results of AZA treatment on relapse frequency and disability. Some preliminary data on brain MRI are also promising. Mycophenolate mofetil (MMF) affects mainly the desired cell types, with a good safety profile, a rapidly reversible activity, and an absence of mutagenic effect and chromosome breakage. However, it remains to be shown that promising experimental results can be converted into significant clinical results in MS. It is presently demonstrated for AZA and it is presumable for MMF that neither drug is able to cure MS. However, it can be anticipated that either drug in combination with other strategies such as recombinant beta interferon could represent a significant adjunct for the therapeutic control of MS, at least in early ambulatory relapsing-remitting MS. Presently, the choice between the old, no longer 'sexy', but well-known drug as AZA and a young, appealing, but still to be better evaluated drug (notably for the long run) as MMF is a matter of personal, community, industrial and scientific inclination. PMID- 9345423 TI - The human Clara cell protein: biochemical and biological characterisation of a natural immunosuppressor. AB - The Clara cell protein, the human counterpart of rabbit uteroglobin, exerts an anti-inflammatory action by interfering in different ways with the cytokine network. Firstly, CC16 behaves like an anti-cytokine by downregulating the production of IFN-gamma, IL-1 and TNF-alpha by stimulated leukocytes. The extent of inhibition depends on the inducing agent (being maximal when IL-2 is used as inducer) and varies with the applied concentration of CC16. Secondly, the protein reduces the antiviral activity and the augmentation of phagocytosis induced by IFN-gamma. In both cases (inhibition of production and biologic activity) there is a 50% reduction in the presence of 10 ng/ml CC16. The natural and IFN-gamma enduced cytotoxicity of NK-cells however, are enhanced by the presence of CC16, indicating a more complex interaction of CC16 with the immune-system. The immunosuppressive properties make CC16 a promising agent for the treatment of inflammatory reactions and auto-immune diseases. PMID- 9345424 TI - Remyelination of the central nervous system. AB - The three typical stages in the clinical course of multiple sclerosis (relapse, persistent disability and progression) can be explained on the basis of inflammation, demyelination and failure of repair leading to axon degeneration and astrocytosis. Strategies are being evaluated for limiting the inflammatory process using immunological treatments and these may have unexpected dividends in promoting endogenous remyelination. Increasing knowledge on glial lineages and axon-glial interactions needed for stable myelination also offer the prospect for enhancing remyelination through growth factor therapy and cell implantation. PMID- 9345425 TI - MS clinical trial design for the future. AB - The Clinical Outcomes Task Force has been challenged to develop new easily administered composite outcomes that are more sensitive, highly reliable, properly validated, and measure more effectively the broad spectrum of independent dimensions of MS. The Task Force on Use of MRI in MS Clinical Trials has already provided important position statements and recommendations for the use of magnetic imaging technologies in MS clinical trials. It appears that TIWGd + activity and change in T2W lesion burden will be most useful as outcomes in patients who recently have experienced frequent relapses. It is anticipated that improved composite outcomes and more powerful statistical methods will facilitate improved predictive validity for MR imaging techniques. If we are successful in meeting these challenges, we should be able to conduct future definitive clinical trials more expeditiously and with fewer patients. PMID- 9345426 TI - Combinations of drugs. AB - As a result of recent therapeutic trials, recombinant Interferon beta (rIFN beta) Ib and -Ia as well as Copolymer I (COP I) and to some extent unspecific immunosuppressants have been accepted as partially efficient treatments of relapsing-remitting MS. In view of partially effective single treatments, the question arises if combination of two or even more-agents could improve efficacy without increasing side effects. The theoretical background of possible combinations is discussed. The selection of combination partners should be based on their proven efficacy as single treatment, on their mode of action and their distinct target in the pathogenetic cascade of events and on their specific side effect profile. Combination regimens selected in this way should undergo evaluation in short term early phase II studies and if the results are positive enter phase III studies. The use of surrogate markers (especially MRI-findings) may help to accelerate this process. Combinations that could enter phase II and III studies in the near future are rIFN beta with unspecific immunosuppressants, e.g. Azathioprine or Methotrexate and rIFN beta with COP I. Combinations including agents just entering phase I and II studies as a single treatment may be more promising for the future. PMID- 9345427 TI - Geographic correlation of multiple sclerosis with tick-borne diseases. AB - An arboviral theory of multiple sclerosis (MS) is presented. Although high MS rates correlate with the distribution of certain population, high rates also correlate with the distribution of Ixodes genus tick viruses. These ticks and viruses are globally distributed by polar-migrating seabirds which are important food sources for island and coastal communities with high MS. Investigation of tick-borne viruses, especially those found in seabirds, in MS is warranted. PMID- 9345428 TI - Interferon beta-1b effects on cytokine mRNA in peripheral mononuclear cells in multiple sclerosis. AB - IFN-beta reduces the number and severity of exacerbations of multiple sclerosis (MS), presumably by modifying immune regulation. We used semiquantitative polymerase chain reaction (RT-PCR) to measure mRNA levels for cytokines before and after IFN beta-1b therapy. mRNA was extracted from mononuclear cells of nine healthy controls and 31 patients with MS. Before therapy, IL-10 and leukemia inhibitory factor (LIF) mRNA levels were elevated in stable MS compared to active MS. Twenty four hours after IFN beta-1b treatment, mRNA levels for IL-1, IL-2, IL 4, IL-6, IL-10, IL-12, IL-13, IFN-gamma, TNF-alpha and LIF had not changed. At 1 week, TNF-alpha mRNA increased and IL-10 and LIF mRNA rose in 75% of patients. IL 2, IL-4, IL-12, IL-13 and IFN-gamma did not change. At 3 months, cytokine mRNA returned to baseline levels. mRNA for the IFN-induced antiviral enzyme, 2,5-OAS, rose by 24 h, peaked at 1 week, and remained elevated thereafter. Serum triglycerides and liver enzymes rose after therapy. Increased SGPT at 3 months correlated with TNF-alpha mRNA levels, suggesting that cytokines may cause some side effects of IFN beta-1b. Baseline cytokine mRNA levels reflect disease activity, but the therapeutic effect of IFN beta-1 b does not appear to be explained by changes in cytokine mRNA levels. PMID- 9345429 TI - T cell response to two immunodominant proteolipid protein (PLP) peptides in multiple sclerosis patients and healthy controls. AB - Multiple sclerosis (MS) is a demyelinating disease of the central nervous system in which autoimmune T lymphocytes reacting with myelin antigens are believed to play a pathogenic role. Since HLA binding is involved in the selection of T cell responses, we have examined PLP peptide binding to HLA DR2, an HLA allele frequently found in MS patients. Both PLP 40-60 and PLP 89-106 show significant, high affinity binding to HLA DR2. We then tested whether responses to PLP peptides 40-60 and 89-106 are elevated in multiple sclerosis patients compared to matched controls. We also analysed T cell responses to MBP 87-106, which is considered to be the immunodominant region of MBP in humans. Here we demonstrate heterogenous T cell responses to PLP 40-60, PLP 89-106 and MBP 87-106 in both MS patients and controls. The overall number of TCL and the HLA restriction of those TCL did not vary significantly in the two groups. PLP 40-60 specific cytolytic TCL were increased in MS patients, whereas healthy controls had increased percentages of cytolytic TCL responding to PLP 89-106 and MBP 87-106. Although the data presented here shows heterogenous responses in T cell numbers, differences in numbers and specificity of cytolytic cells could be involved in the pathogenesis of autoimmune demyelinating disease. PMID- 9345430 TI - Counteracting effect of IFN-beta on IFN-gamma-induced proliferation of human astrocytes in culture. AB - Recent clinical trials have shown that interferon beta (IFN-beta) is effective in reducing exacerbations in relapsing-remitting MS, while interferon gamma (IFN gamma) precipitates the relapses. To investigate mechanisms underlying the beneficial effects of IFN-beta and the detrimental effects of IFN-gamma in MS, cell growth-regulatory effects of IFNs were examined in astrocyte-enriched cultures isolated from fetal brains of 12-20 weeks' gestation. Treatment with IFN gamma (50 or 500 IU ml-1) stimulated significantly the proliferation of astrocytes in 6 out of 9 culture series examined, while IFN-beta (50 or 500 IU ml 1) inhibited the astrocytic proliferation in 3 out of 9 cultures, and IFN-alpha (50 or 500 IU ml-1) did not affect the proliferation IFN-beta and to a lesser degree IFN-alpha reduced the astrocytic proliferation induced by IFN-gamma treatment in 8 out of 9 culture series. The counteracting effect of IFN-alpha/IFN beta against IFN-gamma-induced astrocytic proliferation was verified by the DNA content distribution analysis of propidium iodide-labeled cells. The antagonistic effect of IFN-alpha/IFN-beta on the growth-promoting activity of IFN-gamma in cultured human astrocytes suggests that interferons serve as growth regulators of astrocytes at sites of reactive gliosis lesions of MS. PMID- 9345431 TI - The evolutionary relationship among Caucasian MS patients and controls. AB - Previous observations suggest that the mitochondrial (mt)DNA may confer susceptibility to multiple sclerosis (MS). However, the proportion of affected individuals and the range of contributing mtDNA abnormalities are unknown. To help clarify this question, we analyzed the first hypervariable D-loop sequences of the mtDNA in a group of randomly selected Caucasian MS patients, in MS patients with prominent optic neuritis (PON) and in controls. Phylogenetic analysis of these D-loop sequences revealed that individuals in both groups of patients are generally scattered in the Caucasian phylogeny. However, a small cluster of unrelated MS patients identified by this analysis suggests that a maternal lineage with MS relevant mtDNA sequences may exist, and merits a more comprehensive study. PMID- 9345432 TI - A comparison of clinical and laboratory measures of spasticity. AB - Clinical evaluation of spasticity was performed in lower extremities in 35 ambulatory multiple sclerosis patients and compared with the soleus stretch reflex and the Hoffman reflex. There was no relation between the muscle tone score of dorsiflexion of the foot and the biomechanical/electrophysiological parameters. In contrast, the Achilles tendon reflex score was significantly related to the amplitude (rho = 0.411, P < 0.05) and the slope of the stretch reflex (rho = 0.523, P < 0.01). The clinical examination at the ankle joint revealed 33% normal reflex examinations but only 7% normal muscle tone examinations. In contrast, the number of normal examinations of patellar reflex and muscle tone at the knee joint were similar. It is concluded that the muscle tone score overestimates the amount of spasticity because of changes in the non reflex properties of the spastic extremity and that a reflex score should be used as a clinical measure of spasticity. In addition, biomechanical/electrophysiological evaluation of spasticity at the ankle joint relates to the over-all total muscle tone and reflex scores of lower extremities in this group of MS patients. PMID- 9345433 TI - Characterization of JC virus DNA amplified from urine of chronic progressive multiple sclerosis patients. AB - Thirty-seven chronic progressive multiple sclerosis (MS) patients, 20 of whom were taking cyclosporine, were examined for excretion of JC virus (JCV) in the urine. Polymerase chain reaction (PCR) amplification of DNA in urinary cell extracts detected JCV in 30% of the MS urines. In the cyclosporine treated group four of 20 (20%) excreted JCV, whereas in the untreated group seven of 17 (41%) excreted JCV. Thus, cyclosporine treatment did not enhance urinary excretion of the virus. A control group consisting of an unselected series of 89 patients donating urine in a general medical clinic and 16 healthy volunteers showed 41% with detectable urinary JCV. Thirty-three percent of the control females excreted JCV (18/54), as did 49% of the control males (25/51). Although the percentage of MS patients excreting detectable virus was not increased compared to the control group, the presence of JCV in the urine provides a convenient source of the virus for further characterization. Genotyping of DNA fragments amplified from the VP1 region indicates mainly the presence of JCV Type 1 in these chronic progressive MS patients. This is also the type that predominates in the control group. An apparent recombinant between Type 1 and Type 3 (African) within the VP1 region, tentatively designated Type 1/3 (or Type 4), was found in both the MS group and the controls. A larger series of MS patients that includes relapsing/remitting disease will be required to determine whether the genotype profile of JCV excreted in the urine of MS patients differs significantly from controls. PMID- 9345434 TI - Human T lymphotropic virus type I (HTLV-I)-specific T helper cell responses from HTLV-I seronegative patients with chronic myelopathy and MS in Japan. AB - Human T lymphotropic virus type I (HTLV-I) is a human retrovirus etiologically linked to Adult T cell leukemia (ATL) and HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP). Although most HAM/TSP patients have high anti-HTLV I antibody titers in their sera, HTLV-I infected but seronegative patients with neurological diseases have been reported. To clarify whether seronegative, HTLV-I related neurological disease may exist, we have developed a method that measures the production of interleukin-2 (IL-2) from HTLV-I synthetic peptide-stimulated peripheral blood lymphocytes (PBL) of HTLV-I infected persons. This method is sensitive enough to detect exposure to HTLV-I before seroconversion or even before detection by PCR. We examined 12 patients with chronic progressive myelopathy and eight patients with multiple sclerosis (MS) in central Japan, where the prevalence rate of HTLV-I is between one and four percent among asymptomatic blood donors, using the IL-2 production assay. None of them were positive by the assay, suggesting seronegative HTLV-I myelopathy is very rare among patients with chronic progressive myelopathy and MS in Japan. PMID- 9345435 TI - Failure to detect measles virus RNA, by reverse transcription-polymerase chain reaction, in peripheral blood leucocytes of patients with multiple sclerosis. AB - We have examined peripheral blood leucocytes (PBLs) from 17 multiple sclerosis patients, two patients with rheumatoid arthritis, one case of acute childhood measles and one case of subacute sclerosing panencephalitis, as well as 19 healthy adult controls for measles virus (MV) RNA, by the technique of reverse transcription-polymerase chain reaction. MV nucleocapsid gene specific primers were used to amplify all PBL-derived cDNA samples. These proved to be negative with the exception of the sample derived from the acute measles case. Selected cases were examined further, using fusion gene and matrix gene specific primers. MV RNA could not be detected. PMID- 9345436 TI - Biological assessment and MRI monitoring of the therapeutic efficacy of a monoclonal anti-T CD4 antibody in multiple sclerosis patients. AB - An initial group of 21 patients plus a second group of 14 patients with active multiple sclerosis (MS) (18 progressive and 17 relapsing-remitting forms) were treated with a murine monoclonal anti-T CD4/BF5 antibody as part of a phase I open trial. Tolerance was relatively good: minor general side-effects occurred in 22 patients only upon the first mAb infusion. One year later, functional disability was stabilised in only six of the 35 patients and after 2 years in two patients only (among 21). One year after treatment, nine of the 17 relapsing remitting patients were relapse-free. CD4 counts decreased dramatically 2 h after treatment. These counts were back to baseline counts at 3 months. A transient increase was found in IL-6 and TNF alpha levels 2 h after treatment, which probably accounts for the observed side effects. Cell adhesion molecule levels were not modified. Serial MRI scans were performed in the second group of 14 patients. In all of these patients, lesion modifications were observed in the three scans performed prior to treatment. Yet, no changes in the lesions were noted on the MRI scans performed over the following 3 months. These findings demonstrate the feasibility of this treatment insofar as it induced a marked CD4 lymphocyte depletion. However, it did not seem to stabilise the evolution of the disease--although one must be careful in drawing such conclusions in a phase I trial--or to curb the evolution of MRI-documented lesions. PMID- 9345438 TI - Clinical and magnetic resonance imaging predictors of disability in primary and secondary progressive multiple sclerosis. AB - The role of magnetic resonance imaging (MRI) in predicting disability in multiple sclerosis (MS) remains unclear. In this study 21 patients with primary and secondary progressive MS were reviewed 5 years following a serial MRI study of 6 months duration. In the secondary progressive group (n = 11) there was a significant relationship between the occurrence of enhancing lesions and clinical relapses during the initial 6 months and increase in disability 5 years later. For both groups change in disability over the initial study period was predictive of outcome. These results suggest that the presence and frequency of gadolinium enhancement (a marker of inflammation) and changes in disability over a short period are predictive of future deterioration in progressive patients. PMID- 9345437 TI - Magnetic resonance imaging of epilepsy in multiple sclerosis: a case control study. Implications for treatment trials with 4-aminopyridine. AB - A case-control study of epilepsy in multiple sclerosis (MS) is presented using magnetic resonance (MR) imaging to semiquantitatively assess cortical-subcortical lesion load. In this sample of 13 pairs of cases with MS and epilepsy and controls with MS without epilepsy we found statistically higher cumulated cortical-subcortical lesion loads in the cases than in the controls (Wilcoxon, P = 0.036). Total lesion loads (cortical-subcortical plus deep white matter loads) did not differ significantly (P > 0.1) between cases and controls. The relative risk for seizures as determined by the odds ratio of a cortical-subcortical lesion load of > or = 20 was 8.8 (chi 2 = 5.23, P 0.025), the odds ratio of a large (> 1 cm) cortico-subcortical lesion was 4.7 (chi 2 = 4.9, P < 0.05), while the 2 MR criteria combined show an odds ratio of 19.2 (chi 2 = 8.0, P < 0.005). We conclude that: first, the presence of cortical-subcortical lesions in part accounts for the occurrence of seizures in MS patients; second, due to the substantial overlap of MR imaging scores between cases and controls the ultimate use of these MR imaging findings in the management of individual patients or in the organizations of trials should depend on the expected benefit of the treatment. If the benefit is only moderate or not known a cautious approach with exclusion of cases showing a substantial cortical-subcortical lesion load on MR imaging seems appropriate in trials with drugs, like 4-aminopyridine, that lower the epileptic threshold. PMID- 9345439 TI - Acute unilateral papillitis versus retrobulbar neuritis: relation to multiple sclerosis. AB - Prospectively referred patients with unilateral acute optic neuritis (ON) (n = 223; aged 12-57; 158 women), either idiopathic or part of clinically definite multiple sclerosis (CDMS), were systematically examined by the same physician. We analysed whether the 161 patients with retrobulbar neuritis and the 62 patients with papillitis differed from each other clinically or according to paraclinical tests. The following characteristics were observed in retrobulbar ON respectively papillitis: median age 33 and 33 years, women 70% and 73%, clinically definite MS 30% and 27%. Abnormal results in retrobulbar ON and in papillitis (indicated in brackets) did not differ significantly and were found as follows: cerebral MRI in 56% (63%), VEP from the eye with acute ON in 82% (88%), VEP from the eye without acute ON in 38% (33%), SEP from median nerves in 9% (10%), SEP from tibial nerves in 22% (22%) and biotesiometry in 32% (27%). In the CSF, oligoclonal bands were present in 42% (53%), increased IgG-index in 40% (44%) and increased leucocyte count in 39% (29%). The HLA-DRI5 tissue type was present in 47% (43%). There were no significant differences between retrobulbar ON and papillitis when the idiopathic cases and cases with clinically definite MS were analysed separately. Our data document that unilateral retrobulbar ON and papillitis are both part of the MS spectrum and not different from each other with regard to clinical and paraclinical parameters, indicating that the two groups can be pooled in future treatment trials. PMID- 9345440 TI - Comparison of tirilazad mesylate (U-74006F) and methylprednisolone sodium succinate treatments in experimental allergic encephalomyelitis in the guinea pig. AB - The effects of the non-glucocorticoid 21-aminosteroid, tirilazad mesylate (U 74006F), on MRI and clinical findings in guinea pigs with experimental allergic encephalomyelitis were compared to treatment with methylprednisolone sodium succinate (MPSS). A dose response experiment for U-74006F was performed 1, 3 and 10 mg/kg/day i.p. on day 0-12 after immunization. Additionally, the 3 mg/kg/day i.p. dose was extended to 24 and 35 days. MPSS was given in three different protocols at doses ranging from 0.8 to 3.2 mg/kg/day. Abnormalities in T2 weighted images were assessed as measures of edema and inflammation and gadolinium-DTPA enhanced T1-weighted images were used to determine blood-brain barrier integrity. U-74006F improved the clinical status at doses of 3 and 10 mg/kg. For example, maximum clinical score was halved at 10 mg/ kg/day (P < 0.01). The presence of gadolinium-DTPA in the parenchyma was also decreased at 3 and 10 mg/kg/day U-74006F although maximum MRI scores were decreased only in the 10 mg/kg U-74006F group. Clinical disease suppression seen with 3 mg/kg treatment on days 0-12 reverted to control at > 24 days of dosing. MPSS treatment considerably worsened the clinical outcome of EAE. Mean clinical scores for vehicle and the highest MPSS dose were 0.94 +/- 0.66 versus 2.64 +/- 1.49 (P < 0.05). The combination of decreased T2-weighted abnormalities, clinical signs and gadolinium-DTPA permeation in the U-74006F treated animals suggested protection of the blood-brain barrier without the severe glucocorticoid effects associated with steroid therapy. PMID- 9345441 TI - Myogenic and central neurogenic factors in fatigue in multiple sclerosis. AB - Short episodes of electrical stimulation were applied to the right quadriceps muscle of patients with multiple sclerosis (MS) and healthy subjects at different times during 60 sec sustained voluntary muscle contractions at 0 to 100% levels of maximal voluntarily generated joint torque. The amplitude of electrically induced increments of torque (delta T) has been shown to depend upon both the level of muscular contraction and time from the beginning of the contraction. The dependence of delta T upon the time from the beginning of contraction has been assumed to reflect muscle fatigue. Patients with MS demonstrated an apparent involvement of central neurogenic mechanisms in fatigue manifested as a drop in muscle torque during sustained contractions at 75 and 100% levels when electrical stimulation was able to induce considerable increments in muscle torque. These patients also demonstrated a dependence of delta T upon the contraction level suggesting that they did not produce maximal voluntary contraction torque in the pre-trial. Fatigue in MS is due to central, neurogenic factors and does not seem to involve any myogenic factors such as might be related to secondary muscle changes due to the long-standing disorder. The subjective feeling of tiredness ('fatigue') may be related to a dissociation between central motor commands ('effort') and their mechanical consequences. PMID- 9345442 TI - Viral infection at the blood-brain barrier in multiple sclerosis:--an ultrastructural study of tissues from a UK Regional Brain Bank. AB - Although viral infections are often invoked as environmental factors in the aetiology and pathogenesis of multiple sclerosis (MS) it is only recently that a specific, indirect, cytokine-mediated mechanism for triggering of relapses during viral infections has been demonstrated. It is not yet clear however whether this indirect mechanism can account for all reported viral associations with the aetiopathogenesis of MS. A direct causal role of central nervous system (CNS) viral infection in MS has largely been discounted following repeated failures to demonstrate virus within the oligodendrocyte-myelin unit. In the light of increasing evidence of blood-brain barrier (BBB) dysfunction in MS and to further explore the issue of possible viral involvement in MS, an ultrastructural search for viruses was undertaken in the CNS microvasculature, in autopsy and biopsy tissue from human CNS primary demyelinating diseases, including MS (20 cases), idiopathic monophasic CNS demyelinating disease (Mdemy, four cases) and metabolic or immunopathological demyelinating disease (two cases). For comparison, tissues from CNS viral disease in which demyelination is a major feature (nine cases) were examined in the same way. Control CNS tissues (nine cases) from a range of other neurological and non-neurological diseases were also examined. Outside the MS and Mdemy groups, morphological evidence of virus associations with the BBB were found only in the acute and subacute viral encephalitides (three cases subacute sclerosing panencephalitis, one case of Herpes encephalitis) and in one case of disseminated Cytomegalovirus infection. In a small proportion of MS and Mdemy cases, particles resembling either adenovirus (one case of MS) or paramyxovirus (one case of MS, one case of Mdemy) were found in the vicinity of microvessels. In each case a different cell type or extracellular compartment was involved and an exact correlation between the virus particles and the demyelinating lesions could not be demonstrated. Furthermore, corroborative clinical or laboratory evidence of current CNS infection in these primary demyelinating disease cases was available only from the single positive Mdemy case and not from the two cases of MS. This and other previously published evidence from MS (which implicated a Coronavirus) and other diseases highlights the potential vulnerability to viral infection of cells associated with the BBB. Furthermore it is concluded that the detection rate of such infections in pathological tissue could underestimate their true frequency. A possible role of transient virus-BBB interactions in triggering focal inflammation, BBB breakdown and demyelination in some cases of MS and parainfectious demyelinating disease cannot be discounted. PMID- 9345443 TI - The new partnership in multiple sclerosis: relations with industry. PMID- 9345444 TI - Recent advances in the pharmacological control of experimental allergic encephalomyelitis (EAE) and the implications for multiple sclerosis treatment. AB - The autoimmune, cell-mediated condition experimental allergic encephalomyelitis (EAE) is the representative model for the inflammatory central nervous system disease MS. EAE has been extensively employed to determine the efficacy of pharmacological agents that may be of ultimate use in the treatment of MS. A wide variety of drugs has been examined for activity in EAE but, over the last decade, three groups of compounds have emerged with clear and reproducible ability to modify significantly the onset and progression of the disease. The immunosuppressants, the modulators of catecholamine activity and the antineoplastic agents have convincingly altered the course of EAE and, as a consequence, provided understanding of the mechanisms of disease expression and offered further insight into the pathogenesis of MS. The article stresses the usefulness of EAE as a model to identify prospective pharmacological treatments for MS and, in particular, considers those compounds subsequently assessed for their ability to interfere with the progression of the human disease. PMID- 9345445 TI - Multiple sclerosis progression in a natural history study: predictive value of cerebrospinal fluid free kappa light chains. AB - OBJECTIVES: To determine the relationship between baseline CSF immunologic abnormalities and MS disease progression; To determine progression rates in an untreated, recently-diagnosed MS sample using several validated clinical measures. BACKGROUND: CSF immune abnormalities are common in MS but have not been linked to disease progression. DESIGN METHODS: Thirty-six patients with definite (n = 28), probable (n = 2), or possible (n = 6) MS were studied prospectively. Baseline CSF was analyzed for free kappa and lambda light chains, myelin basic protein, IgG synthesis rate, and IgG index. MS patients were entered into the study within 5 years of symptom onset and examined semiannually for as long as 53.4 months (median length of follow up 38.9 months). MS progression was defined as sustained worsening on the following clinical measurement instruments: the Expanded Disability Status Scale (EDSS), the Ambulation Index (AI), the 9 Hole Peg Test (9HT) and the Box and Blocks test (BBT). Kaplan-Meier estimates of disease progression were calculated and the relationship between baseline CSF values and disease progression was determined using Cox proportional hazards regression models. RESULTS: By 36 months, 33% (95% CI = 10.3, 55.7) of patients had progressed on EDSS and 49.7% (95% CI = 27.7, 71.7) had progressed on at least one of the outcome measures. Patients with CSF free kappa chain levels in the upper quartile had a significantly higher risk of progression on EDSS (Hazard Ratio 3.78; p = 0.04) and 9HT (Hazard Ratio 10.77, p = 0.04). CONCLUSIONS: In this study, CSF free kappa light chains predicted subsequent physical deterioration in prospectively evaluated MS patients. If this is confirmed by larger studies, then CSF free kappa light chains could serve as a target for intervention in therapeutic trials. PMID- 9345446 TI - Cardiovascular regulation in multiple sclerosis. AB - Traditional assessments of autonomic nervous system function have depended on invasive and complex procedures. Vagal power, which is the respiratory component of heart rate variability (HRV) is an alternative and non-invasive measure for indexing autonomic nervous control of the heart. In the current study, 18 multiple sclerosis (MS) and 20 healthy subjects matched with respect to age, education and intelligence served as subjects. The MS group showed significantly lower vagal power during natural and paced breathing than healthy subjects. Importantly, heart rate did not differ between the two groups. If MS patients exhibit abnormalities in mechanisms mediating cardiac parasympathetic control, the impact on quality of life and vulnerability to adverse cardiac events need to be further evaluated. The results of this study may have implications with respect to the feasibility of using HRV as both a diagnostic and prognostic tool for evaluating parasympathetic nervous system dysfunction and in providing valuable information for developing more effective treatment and rehabilitation strategies. PMID- 9345447 TI - Laryngeal Uhthoff's phenomenon: a case report. AB - An Uhthoff-like phenomenon was recently observed in a patient with clinically definite MS who experienced transient dysphonia brought on by exertion and relieved by cooling. The patient's dysphonia was felt to be related to intermittent temperature-dependent conduction block associated with a demyelinating plaque in the region of the left nucleus ambiguus. We have termed the patient's intermittent dysphonia 'laryngeal Uhthoff's phenomenon'. PMID- 9345448 TI - The Brief Repeatable Battery of Neuropsychological Tests for Multiple Sclerosis: a preliminary serial study. AB - The Brief Repeatable Battery of Neuropsychological Tests (BRB-N) for MS consisting of the selective reminding, 10/36 spatial recall, symbol digit modalities, paced auditory serial addition (PASAT) and word list generation tests is a sensitive measure of early cognitive impairment in MS patients. We administered it to 19 chronic stable MS patients every 60 days for 120 days to examine variability. The mean coefficient of variation for the tests ranged from 18% to 22%. A significant practice effect was seen in the PASAT results (P < 0.05) using the Wilcoxon signed rank test. These results suggest that cognitive fluctuations analogous to motor fluctuations may occur in MS patients and that the BRB-N may be useful in clinical trials of agents expected to alter cognitive function in MS patients if test-retest variability and practice effects are taken into account. Further study is warranted. PMID- 9345449 TI - Clinical and subclinical neurological involvement in children of conjugal multiple sclerosis patients. AB - We report the occurrence of clinically definite multiple sclerosis in an offspring of a couple with conjugal multiple sclerosis. Extensive investigation of all members of this family, which includes two additional asymptomatic children, eliminated the possibility of alternative neurological diagnoses. All family members were studied with magnetic resonance imaging (MRI), evoked potentials, and human leukocyte antigen (HLA) typing. An asymptomatic child had subtle white matter abnormalities on MRI, suggesting subclinical neurological involvement. This study documents the third case of multiple sclerosis in the child of conjugal multiple sclerosis patients and provides the first report of MRI lesions in an asymptomatic offspring of the same parents. Neurodiagnostic and immunogenetic investigations of such rare family clusters may contribute to the elucidation of the pathogenesis of multiple sclerosis. PMID- 9345450 TI - Expression of IFN-gamma, IL-4, and TGF-beta in multiple sclerosis in relation to HLA-Dw2 phenotype and stage of disease. AB - Multiple sclerosis (MS) is associated with upregulation of both proinflammatory (interferon-gamma, IFN-gamma) and immunosuppressive (transforming growth factor beta, TGF-beta) cytokines. To examine a possible relation between the MS-related HLA haplotype Dw2 and cytokine profiles, we used in situ hybridization with labeled cDNA oligonucleotide probes to detect transcripts of the T helper type 1 (Th 1) cell related IFN-gamma, the Th2 cell related interleukin-4 (IL-4) and of TGF-beta in blood and cerebrospinal fluid (CSF) mononuclear cells from 62 patients with MS. Compared to patients with other neurological diseases and healthy controls, MS patients had elevated numbers of IFN-gamma, IL-4 and TGF beta mRNA expressing cells in blood and further augmented in CSF. Although several HLA-Dw2-positive individuals showed very high numbers of cells expressing these cytokines, no significant difference was found in comparison with Dw2 negative patients. However, expression of IL-4 and TGF beta mRNA was significantly increased in patients with shorter duration and minor disability and, for IL-4, in patients still in the relapsing-remitting phase compared to patients with secondary chronic progressive MS. Surprisingly, these changes which favour a beneficial, disease-downregulating effect of IL-4 and TGF-beta in MS, were found to be confined to HLA-Dw2-positive patients. Our findings suggest that the HLA phenotype does not influence the overall level of immune reactivity in MS, but may distinguish subgroups characterized by particular cytokine expression patterns. PMID- 9345451 TI - An evaluation of tumor necrosis factor microsatellite alleles in genetic susceptibility to multiple sclerosis. AB - We have analyzed the distribution of tumor necrosis factor (TNF) a and -b microsatellite alleles in HLA-DQ and -DR typed Swedish patients with multiple sclerosis (MS) (n = 122) and ethnically matched control subjects (n = 178). We found significant differences in the frequencies of TNFa and TNFb alleles between patients and controls. TNFaII was significantly associated with MS. This was also the case for the combination of TNFaII with TNFb4. However, TNFaII (alone or in combination with TNFb4) did not show any disease association independent of DQA1*0102/ DQB1*0602/DR2, whereas the previously reported strong association with HLA-DQA1*0102/DQB1*0602/DR2 in Scandinavian populations was confirmed. Therefore the association of TNFaII (and TNFb4) is most likely secondary to the increase of DQA1*0102/DQB1*0602/DR2 in MS patients. The proportion of TNFa6 positive individuals was lower among DR2-negative MS patients than among DR2-negative controls (P = 0.08). Since the presence of the TNFa6 allele correlates with low TNF alpha production in response to lipopolysaccharide, it could be speculated that DR2-negative MS patients have an increased risk of being high TNF alpha producers in response to exogenous stimuli. PMID- 9345452 TI - PCR typing of two short tandem repeat (STR) structures upstreams of the human myelin basic protein (MBP) gene; the genetic susceptibility in multiple sclerosis and monosymptomatic idiopathic optic neuritis in Danes. AB - We investigated two short tandem tetranucleotide (TGGA) repeat polymorphisms upstreams of the myelin basic protein (MBP) gene. The region was amplified by the polymerase chain reaction (PCR) and the two repeat systems were separated by cutting with the restriction enzyme NlaIII. The lengths of the DNA fragments were analyzed by vertical electrophoresis in polyacrylamide gels followed by silver staining. We compared the DNA fragment frequencies of the two MBP regions in 34 patients suffering from multiple sclerosis and in 78 suffering from monosymptomatic idiopathic optic neuritis to those in 200 healthy controls. We found no significant differences between the MBP fragment frequencies in either of the patient groups and in the control group. PMID- 9345453 TI - Viruses and multiple sclerosis. PMID- 9345454 TI - Mechanisms of damage and repair in multiple sclerosis--a review. AB - Pathological features of MS include perivascular inflammation and demyelination with oligodendrocyte loss; in addition, attempts at remyelination are often unsuccessful and may culminate in astrocytic scarring. One approach to investigating the biological principles underlying these processes is to use in vitro systems to analyse single-cell behaviour as well as cell-cell interactions. This paper reviews such data concerned with cell injury and repair which illuminate both demyelination and remyelination. In tissue culture oligodendrocytes are susceptible to injury via cell-mediated and humoral mechanisms. Substances including complement and tumour necrosis factor are capable of killing rat oligodendrocytes in vitro; surface complement activation also initiates a number of intracellular processes within oligodendrocytes as well as providing ligands for phagocytic interactions. The reasons for oligodendrocyte complement activation are discussed, but it appears that species differences exist when extrapolating these data to humans. Myelination and remyelination can also be studied both in vitro and in vivo using defined cell populations. Results from these studies may eventually help to explain some pathological features of MS, including astrocytosis and factors governing the limits of remyelination. PMID- 9345455 TI - Increased risk of multiple sclerosis after late Epstein-Barr virus infection: a historical prospective study. AB - An association between infectious mononucleosis (IM) and MS has been proposed. In a historical prospective study we used records from the Danish State Serum Institute on heterophile antibody (HA) tests for IM performed in all Danish patients over a number of years. Included in the analysis were 6853 HA-positive persons analyzed from 1968 to 1978 (except 1975) and 12,886 HA-negative per sons analyzed in the years 1968, 1969, 1970 and 1978. A search for these persons in the central nationwide Danish Multiple Sclerosis Registry (DMSR) was performed. Among the HA-positive persons 16 cases of MS which met the diagnostic criteria were found with onset of MS after the year of the HA test and before follow-up on 1 January 1991. The expected number for a Danish population, matched by sex, age and year at start of observation, was 5.70 (P < 0.05), the risk ratio being 2.81. No patient had developed MS before contracting IM. Among the HA-negative persons 12 were registered with onset of MS after the year of the HA test and before follow-up, the expected number being 10.47 (P > > 0.05). Although Epstein-Barr virus is not suggested in itself to be the cause of MS, we propose that it is a co-factor in the pathogenesis of this disease. PMID- 9345456 TI - B-lymphoblastoid cell lines from multiple sclerosis patients and a healthy control producing a putative new human retrovirus and Epstein-Barr virus. AB - On several occasions we have observed retrovirus-like particles (RVLPs) by transmission electron microscopy (EM) of cultured T cells from a patient with MS. Later we established spontaneously formed B-lymphoblastoid cell lines (LCLs) from a patient with an MS-like disease and from another patient with MS who had a reactivated Epstein-Barr virus (EBV) infection. Both LCLs were found by EM to produce RVLP and EBV particles. Reverse transcriptase (RT) assays were positive in purified viral material from both LCLs. To substantiate these findings we initiated an intensified culturing procedure and were able to establish LCLs from 5 out of 21 consecutive MS patients and 1 out of 13 consecutive healthy controls. All LCLs were found to produce both RVLP and EBV particles by EM. Whether the putative new retrovirus(es) and EBV have any causal relationship to MS is still not known, but the findings support this possibility. PMID- 9345457 TI - Expression of endogenous retroviruses in blood mononuclear cells and brain tissue from multiple sclerosis patients. AB - The aim of the present study was to examine whether there is an abnormal expression of certain endogenous retroviruses in MS patients. For this purpose samples of peripheral blood mononuclear cells were obtained from 22 MS patients, a corresponding number of age and sex-matched healthy donors and five patients with other diseases affecting the central nervous system. In addition, brain specimens of macroscopic normal white and gray matter from four MS patients and a similar number of controls were included in the study. Using an enzymatic amplification technique, we found expression of the endogenous retroviral sequences, HRES-1, HERV-K10 and ERV3 in most samples of peripheral blood mononuclear cells from MS patients and controls without obvious differences between these two groups. In contrast, composite transcripts of ERV3 and a zinc finger sequence were more frequently detected in healthy donors than in MS patients. At present, the possible significance of this is uncertain. The retroviral element 4-1 was not transcribed or only transcribed at a very low level in peripheral blood cells of controls and MS patients. Transcripts of various endogenous retroviruses were also detected in the brain samples, but a different pattern was not apparent in the MS group as compared with controls. Aspects concerning a possible association between endogenous retroviruses and autoimmunity are considered. PMID- 9345458 TI - Is LM7 a new human retrovirus or a mycoplasma virion? AB - A putative retrovirus called LM7 was recently isolated from a patient with MS. This retrovirus was detected in LM7 and LM711 cultured human leptomeningeal cells. In the present work, nucleic acids from LM711 cell culture supernatants were purified and subjected to avian myeloblastosis viral (AMV) reverse transcriptase and to random polymerase chain reaction (rPCR) in order to characterize the genomic material of LM7 virions. Analysis of reverse transcription products allowed the detection of an approximately 14 kb ribonucleic acid in all LM711 cell culture supernatants. However, sequencing of rPCR-amplified molecules as well as RNA blotting data showed essentially that all tested cells producing LM7 particles were infected with mycoplasma. Moreover, purification of LM7 particles onto a linear sucrose density gradient established that the 14 kb nucleic acid was always associated with the 1.19-1.21 g ml-1 sucrose fractions, which are known to correspond to the buoyant density of mycoplasma. In addition, no viral genomic RNA was detected in the 1.17 g ml-1 sucrose fraction containing the low reverse transcription activity. These results strongly suggest that microscopic images and serological data could be related to mycoplasma and/or to a virion associated with the bacteria. The LM7 particle might be a new and additional enveloped virus able to infect Mycoplasma hyorhinis hosts. Thus, for instance, it would be presumptuous to assert, with our current understanding, that the LM7 virion is one of the causal agents of MS in humans. PMID- 9345459 TI - A hybrid between a resistant and a susceptible strain of mouse alters the pattern of Theiler's murine encephalomyelitis virus-induced white matter disease and favors oligodendrocyte-mediated remyelination. AB - Theiler's murine encephalomyelitis virus (TMEV) produces a chronic disease in its natural host, the mouse, characterised by primary inflammatory demyelination of the spinal cord. This viral infection is considered a very good model for human MS because the pathogenesis of myelin injury is mediated through the host immune response. Susceptibility and/or resistance to the demyelinating disease depend on multiple genes both in and outside the major histocompatibility complex (MHC). The pathological lesions in animals with different degrees of susceptibility vary in both their severity and in their ability to become remyelinated. In general, animals with intermediate levels of susceptibility show the best potential for remyelination. Most crosses of susceptible animals with resistant strains carrying the H-2b haplotype are resistant with only a couple of exceptions. One such exception is the (SJL/J x C57L/J)F1 hybrid, which is susceptible to the disease. To study whether the resistant genotype of C57L/J mice could modify the phenotypic expression of pathological lesions characteristic of the highly susceptible SJL/J mouse, we performed a light microscopical and ultrastructural study of the spinal cord of both parental strains and their F1 progeny. We focused particularly on the relationship between severity of inflammation, and especially macrophage infiltration, and the subsequent remyelinating potential of lesions. The results show a dramatic difference between the ability to remyelinate lesions by infected SJL/J mice vs similarly infected (SJL/L x C57L/J)F1 hybrids, and suggest an important influence by resistant genes in modulating the phenotypic expression of disease, including the ability to stimulate oligodendroglia-mediated remyelination. PMID- 9345460 TI - Depression before and after diagnosis of multiple sclerosis. AB - Depression was examined in 45 patients evaluated within 2 months of diagnosis of MS. At the time of testing, 40% of the MS sample met the diagnostic criteria for major depression, 22% had adjustment disorder with depressed mood and 37% showed no evidence of mood disorder. Personal and family history of depression in patients with MS was also examined and compared with a sample of patients with chronic low back pain (CLBP) who were matched for age, gender, marital and employment status and current level of depression. Fifty-two per cent of patients with MS reported experiencing a depressive episode before the onset of MS compared with 17% of patients with CLBP (P < 0.001). Sixteen patients with MS (35%) reported family history (parent or sibling) of treatment for depression compared with seven (15%) of patients with CLBP (P < 0.05). MS patients with a history of depression reported more initial symptoms than MS patients without a history of depression. Clinical and theoretical implications of the findings are discussed. PMID- 9345461 TI - Experimental allergic encephalomyelitis in non-human primates: diffusion imaging of acute and chronic brain lesions. AB - Diffusion imaging and T2-weighted magnetic resonance imaging were performed on 16 monkeys with experimental allergic encephalomyelitis (EAE), a model of the human demyelinating disease MS. The purpose of this study was to determine whether local changes in diffusion image intensity could be correlated with the formation of acute and chronic demyelinating lesions. Diffusion image analysis was restricted to the internal capsule of the brain because of its anatomic orientation of fiber pathways. Acute inflammatory EAE lesions were large and monophasic, as visualized by T2-weighted MRI, and were accompanied by a decrease in the diffusion MR image signal with the diffusion-sensitizing gradient in all three orthogonal directions (n = 27 brain regions, P < 0.005). Chronic demyelinating lesions were preceded by multiple inflammatory attacks, as visualized by MRI, and by a decrease in diffusion MR image signal with the diffusion-sensitizing gradient in the two orthogonal directions perpendicular to the fibers of the internal capsule (n = 18 brain regions, P < 0.005). However, for the chronic group, there was no significant change in the diffusion MR image signal with diffusion-sensitizing gradient parallel to the fibers of the internal capsule at the terminal scan, suggesting little change in the water diffusion within the nerve fibers. These results suggest that diffusion imaging holds promise for measuring subtle changes in water diffusion due to different types of brain damage. PMID- 9345462 TI - A phase III trial of intramuscular recombinant interferon beta as treatment for exacerbating-remitting multiple sclerosis: design and conduct of study and baseline characteristics of patients. Multiple Sclerosis Collaborative Research Group (MSCRG). AB - The design and conduct of a randomized, double-blinded, placebo-controlled, multicenter, phase III study of recombinant interferon beta-1a (IFN-beta-1a) as treatment for exacerbating-remitting MS are described, as are baseline characteristics of the study population. The purpose of the study was to determine if 6.0 x 10(6) IU (30 micrograms) of IFN-beta-1a, administered by weekly intramuscular (i.m.) injections, was effective in delaying the onset of sustained disability. The primary outcome measure was time to onset of treatment failure, defined as a worsening on the Kurtzke Expanded Disability Status Scale (EDSS) of greater than or equal to 1.0 point compared with baseline, persisting for at least 6 months. An intent-to-treat design was used. The primary outcome measure was analyzed using the Mantel-Cox log-rank statistic and Kaplan-Meier survival curves. Secondary outcomes included quantitative measures of upper and lower extremity function, neuropsychological test performance, functional and quality of life assessments and several measures derived from annual brain MRI studies. Entry criteria included prestudy exacerbation rates of at least 0.67 per year and EDSS scores of 1.0-3.5. A total of 301 MS patients were randomly assigned to receive weekly i.m. injections of IFN-beta-1a or placebo. The average age of the study population at entry was 37 years; 92% were Caucasian and 73% were women. The mean prestudy disease duration was 6.5 years, mean prestudy exacerbation rate was 1.2 per year and the mean EDSS score was 2.3. The randomization yielded well-balanced treatment arms. Various aspects of the study are discussed, including: (1) the decision to focus study design on sustained disability; (2) the rationale for the treatment regimen; (3) measures taken to assure the reliability of the primary outcome measure; and (4) a description of the secondary outcome measures. PMID- 9345463 TI - Insulin-like growth factor I treatment reduces clinical deficits and lesion severity in acute demyelinating experimental autoimmune encephalomyelitis. AB - Our goal was to test the effects of insulin-like growth factor I (IGF-I) treatment on clinical deficits, lesion number and lesion size in acute demyelinating experimental autoimmune encephalomyelitis (EAE) induced in Lewis rats with an emulsion containing guinea pig spinal cord. In this EAE model, there is severe immune-mediated demyelination, which resembles that seen in actively demyelinating MS lesions. On day 12-13 after EAE induction, a total of 23 pairs of rats with the same mild degree of tail and hind limb weakness were given either intravenous IGF-I or placebo twice daily for 8 days. The daily IGF-I dose used in the first trial was 200 micrograms (about 0.6 mg kg-1) and in the second and third trials was 1 mg (about 3.0 mg kg-1). IGF-I treatment reduced permeability of the blood-spinal cord barrier to Evans blue-albumin. Maximum clinical deficit scores of IGF-I-treated rats were significantly lower and treated rats recovered faster than controls. IGF-I treatment produced significant reductions in weight loss and hind limb weakness. Treatment also improved treadmill walking, stride length and climbing performance. Morphometric analysis showed that spinal cord inflammatory lesions were significantly smaller and fewer in IGF-I-treated rats. The higher IGF-I dose produced a greater reduction in clinical and pathological deficits. We conclude that IGF-I treatment promotes clinical recovery by reducing EAE-induced blood-spinal cord barrier changes and the associated immune-mediated inflammatory lesions. Our results suggest that IGF I may be useful in treating patients with multiple sclerosis and other demyelinating diseases. PMID- 9345464 TI - Immune and non-immune actions of interferon-beta-Ib on primary human neural cells. AB - Systemic interferon-beta-Ib (IFN-beta-Ib) reduces the frequency of clinical exacerbations and the number of magnetic resonance imaging (MRI)-defined lesions in patients with relapsing-remitting MS. The basis for this clinical effect is not understood. While IFN-beta-Ib has been demonstrated to have antiproliferative and immunomodulatory effects on the systemic immune system, its actions on neural cells could also contribute to its therapeutic efficacy. In this study, we have examined possible immune and non-immune effects of IFN-beta-Ib on CNS-derived primary human cells. With respect to immune-related effects, application of IFN beta-Ib did not decrease basal expression of HLA-DR on astrocytes or microglia, and it reduced the IFN-gamma-enhanced HLA-DR expression on adult human astrocytes only at high concentrations (1000 IU ml-1); IFN-beta-Ib at all concentrations tested did not reduce the IFN-gamma-enhanced HLA-DR expression by fetal astrocytes or adult microglial cells. In contrast, but in correspondence with the literature, the IFN-gamma-enhanced HLA-DR expression on a glioma cell line was attenuated by IFN-beta-Ib in a dose-dependent manner. With respect to non-immune effects, the number of adult human oligodendrocytes and their state of morphological differentiation were not affected by IFN-beta-Ib. Proliferation of the mitotically active fetal human astrocytes, however, was reduced by IFN-beta Ib treatment. Lactate dehydrogenase assays revealed that IFN-beta-Ib was not toxic to neural cells, including adult oligodendrocytes and fetal human neurons. We conclude that IFN-beta-Ib lacks efficacy in down-regulating HLA-DR expression by primary human neural cells and that regulation of MHC class II antigens is unlikely to be a mechanism for its beneficial effect in MS. Finally, the lack of toxicity of IFN-beta-Ib on human neural cells is important for a drug that will probably be used widely. PMID- 9345465 TI - Transient immunomodulation by intravenous methylprednisolone treatment of multiple sclerosis. AB - The extent and duration of immunomodulation induced by high-dose corticosteroid treatment of clinical relapse of multiple sclerosis was investigated. Ten patients treated with a 5 day course of intravenous methylprednisolone (IVMP) (500 mg daily) were studied. Circulating lymphocyte subpopulations and mitogen induced interleukin 2 (IL-2) and gamma-interferon (gamma-IFN) production were determined immediately before initiation of therapy (day 1), during therapy (24 h after first dose, day 2) and at 24 h and 1 week post therapy (days 6 and 12 respectively). T-cell subpopulation (CD3, CD4, CD8, CD4CD45RA, CD4CD45RO) levels fell within 24 h of initiation of therapy, rebounded above pretreatment levels at day 6 and normalised 1 week post therapy. Despite a reduction in total T-cell numbers during treatment, the gamma delta T-cell subpopulation was not significantly altered. HLA-DR expression on B cells and monocytes declined transiently on day 2 to approximately 50% of pretherapy levels. IL-2 and gamma IFN production were reduced during therapy but returned to baseline levels by 24 h post therapy. The effects of IVMP on lymphocyte distribution and function appear to be short-lived and, therefore, may not be responsible for the rapid improvement associated with this form of treatment. PMID- 9345466 TI - A study of multiple sclerosis patients with magnetic resonance spectroscopic imaging. AB - Eleven patients with clinically definite MS and three healthy controls were investigated by magnetic resonance spectroscopic imaging. The data sets were analysed for all voxels containing white matter only. We classify these voxels in healthy controls as normal white matter (NWM), and in MS patients as normal appearing white matter unaffected by MS lesions (NAWM) or white matter affected by MS lesions. The spectra belonging to the voxels were analysed for content of cholines, creatines and N-acetylaspartate (NAA), and compared as a group. It was found that lesions differ from white matter in chemical composition and, moreover, that normal-appearing white matter differs from healthy white matter. Specifically, levels of NAA are lower in patients. There seems to be a linear relation between the composition of white matter and the expanded disability status scale value for the patient. The presence of lactate could not be established, and no unambiguous differences were found between patients with relapsing-remitting and relapsing-progressive disease. PMID- 9345467 TI - Mitochondrial DNA mutations in multiple sclerosis. AB - The presence of mitochondrial DNA mutations, including eight of those frequently associated with Leber's hereditary optic neuropathy (LHON), was investigated by sequencing and restriction endonuclease analysis in randomly selected patients with MS. Class I LHON mutations with primary pathogenic significance for blindness were not detected in any of the MS patients studied. A trend was observed for higher frequency of class II LHON mutations with unknown pathogenic significance in the MS patients than in the controls. Specifically, the mutation at position 4216 and its associated simultaneous mutations occurred with a higher frequency. Eleven of the 53 patients (20.8%) were positive for at least two (4216 and 4917 or 13,708) or three (4216, 13,708, 15,257) simultaneous class II LHON mutations, while 7 of the 74 controls (9.5%) carried simultaneous mutations (P = 0.036). Earlier studies reported the occurrence of either the 11,778 or 3460 LHON type mutations in MS patients with a positive LHON pedigree and/or with a disease course predominantly involving the optic nerves. The mutations we detected did not correlate with the severity of visual loss in either LHON or MS, rather they seemed to be present in randomly selected MS patients. We conclude that the mutations with primary pathogenic significance for blindness are not shared between LHON and randomly selected MS. However, the presence of further mitochondrial mutations cannot be excluded in MS. The increased incidence of the simultaneous class II LHON mutations in MS patients (and LHON) vs controls may indicate that certain sets of mitochrondrial DNA mutations/variants are associated with and predispose to MS, a possibility which needs to be investigated further. Alternatively, a biological disadvantage may be associated with the coexistence of the mutations detected. PMID- 9345468 TI - Outcomes assessment in multiple sclerosis clinical trials: a critical analysis. AB - The feasibility and precision of clinical trials for the treatment of MS must be improved. Subsequent to the approval by the Food and Drug Administration of the United States of interferon beta-Ib as a safe and effective, though not curative, treatment for relapsing-remitting MS, the testing of other agents in this disease has been undertaken or is anticipated. This report summarises the discussions and recommendations of an international workshop held to review critically the elements of current MS therapeutic trials and to identify the most important aspects of clinical evaluation, study design and data analysis that would allow agents for MS to be tested as accurately, rapidly and economically as possible. While acknowledging the many uncertainties about the pathophysiology and natural history of MS, the workshop participants made recommendations about the preferred components to be used in the design of trials which may be different depending on the treatment goal and agent studied. It was concluded that the formulation of a useful clinical trial design must be based on specific guidelines for clinical scales and imaging for which task forces were recommended and subsequently appointed. PMID- 9345469 TI - Case finding for epidemiological surveys of multiple sclerosis in United States communities. AB - In regard to prevalence surveys of MS in US communities, this article considers the relative merits of case finding through hospitals, physicians, MS service organisations, neurology practices, death certificates, chronic care facilities and media announcements and lay referrals. The current value of hospitals and non neurological practitioners for case finding is questionable, given the changes in clinical practice with respect to MS and the data collection problems inherent with surveys of physicians. We suggest giving priority to the following case finding methods: manually reviewing patient records of neurology practices, screening patient rolls of local MS service organisations and surveying chronic care facilities. In some US communities, these methods may need to be augmented by also surveying internists, general practitioners and family practitioners. Capture-recapture methodology offers a way to evaluate the completeness of case ascertainment. PMID- 9345470 TI - Locating the motor cortex on the MRI with transcranial magnetic stimulation and PET. AB - Transcranial magnetic stimulation with a focal coil was used to map the cortical representation of a hand muscle in four healthy subjects. In each subject, the three-dimensional locations of the magnetic stimulation positions and about 400 positions on the surface of the head were digitized. The amplitude-weighted center of gravity of each subject's map was found, and a line perpendicular to the local head surface was projected inward. The digitized heads were registered with the subjects' MRIs using the scalp contours. The coordinate transformations yielded by this process were used to map the stimulation positions and the perpendicular line into the MRIs. Brain areas imaged with positron emission tomography (PET) and 15O-labeled water, activated by movement of the same muscle, were registered with the MRIs using the brain contours. In all cases, the magnetic stimulation lines encountered the surface of the brain at the anterior lip of the central sulcus and ran along the precentral gyrus a few millimeters anterior to the central sulcus, coming within 5-22 mm of all the PET activation maxima. This technique demonstrates the accuracy of transcranial magnetic stimulation for locating the primary motor area. PMID- 9345471 TI - Improved methods for image registration. AB - We report a system for PET-MRI registration that is improved or optimized in several areas: (1) Automatic scalp/brain segmentation replaces manual drawing operations, (2) a new fast and accurate method of image registration, (3) visual assessment of registration quality is enhanced by composite imaging methods (i.e., fusion) and (4) the entire procedure is embedded in a commercially available scientific visualization package, thereby providing a consistent graphical user interface. The segmentation algorithm was tested on 17 MRI data sets and was successful in all cases. Accuracy of image registration was equal to that of the Woods algorithm, but 10 times faster for PET-PET and 4 times faster for PET-MRI. The image fusion method allows detection of misalignments on the order of 2-3 mm. These results demonstrate an integrated system for intermodality image registration, which is important because the procedure can be performed by technicians with no anatomic knowledge and reduces the required time from hours to about 15 min on a modern computer workstation. PMID- 9345472 TI - High-resolution random mesh algorithms for creating a probabilistic 3D surface atlas of the human brain. AB - Striking variations exist, across individuals, in the internal and external geometry of the brain. Such normal variations in the size, orientation, topology, and geometric complexity of cortical and subcortical structures have complicated the problem of quantifying deviations from normal anatomy and of developing standardized neuroanatomical atlases. This paper describes the design, implementation, and results of a technique for creating a three-dimensional (3D) probabilistic surface atlas of the human brain. We have developed, implemented, and tested a new 3D statistical method for assessing structural variations in a data-base of anatomic images. The algorithm enables the internal surface anatomy of new subjects to be analyzed at an extremely local level. The goal was to quantify subtle and distributed patterns of deviation from normal anatomy by automatically generating detailed probability maps of the anatomy of new subjects. Connected systems of parametric meshes were used to model the internal course of the following structures in both hemispheres: the parieto-occipital sulcus, the anterior and posterior rami of the calcarine sulcus, the cingulate and marginal sulci, and the supracallosal sulcus. These sulci penetrate sufficiently deeply into the brain to introduce an obvious topological decomposition of its volume architecture. A family of surface maps was constructed, encoding statistical properties of local anatomical variation within individual sulci. A probability space of random transformations, based on the theory of Gaussian random fields, was developed to reflect the observed variability in stereotaxic space of the connected system of anatomic surfaces. A complete system of probability density functions was computed, yielding confidence limits on surface variation. The ultimate goal of brain mapping is to provide a framework for integrating functional and anatomical data across many subjects and modalities. This task requires precise quantitative knowledge of the variations in geometry and location of intracerebral structures and critical functional interfaces. The surface mapping and probabilistic techniques presented here provide a basis for the generation of anatomical templates and expert diagnostic systems which retain quantitative information on intersubject variations in brain architecture. PMID- 9345473 TI - Autoradiographic evidence that 3-quinuclidinyl-4-fluorobenzilate (FQNB) displays in vivo selectivity for the m2 subtype. AB - Alzheimer's disease (AD) involves selective loss of muscarinic m2, but not m1, subtype neuroreceptors in cortical and hippocampal regions of the human brain. Emission tomographic study of the loss of m2 receptors in AD is limited by the fact that there is currently no available m2-selective radioligand which can penetrate the blood-brain barrier. We now demonstrate the in vivo m2 selectivity of a fluorine derivative of QNB (FQNB), by studying autoradiographically the in vivo inhibition of radioiodinated (R)-3-quinuclidinyl (S)-4-iodobenzilate ((R,S) [125I]IQNB) binding by unlabeled FQNB. In the absence of FQNB, (R,S)-[125I]IQNB labels brain regions in proportion to the total muscarinic receptor concentration; in the presence of 30.0 nmol of racemic FQNB, (R,S)-[125I]IQNB labeling in those brain regions containing predominantly the m2 subtype is reduced to background levels. We conclude that FQNB is m2-selective in vivo and that [18F]FQNB or a closely related analogue may be of potential use in positron emission tomographic study of the loss of m2 receptors in AD. PMID- 9345474 TI - The effect of varying stimulus rate and duration on brain activity during reading. AB - The effect of the presentation rate and exposure duration of visually presented words on brain activity was investigated using positron emission tomography. Subjects either read aloud or silently mouthed the names of words. In regions associated with early visual analysis, activity increased with both rate and duration; in regions associated with response generation, activity increased with increasing rate but was unaffected by duration; and in regions associated with word recognition, activity decreased with increasing duration. The variable responses of different brain regions illustrate the functional segregation of these regions. Of particular interest was the dissociation between activity in the posterior fusiform gyri and that in the medial lingual gyrus--in the former, activity increased with rate and duration but the latter was unaffected by either variable. This finding suggests that word processing in the lingual gyrus during reading is distinct from that in the posterior fusiform gyri. A further observation was that during reading aloud, when subjects can hear the sound of their own voice, the response in the primary auditory cortices increased with stimulus rate, demonstrating that subjects process the sound of their own voice in a qualitatively similar way to words spoken by another. PMID- 9345475 TI - Cortical activation in the human brain during lateral saccades using EPISTAR functional magnetic resonance imaging. AB - The location of the human cortical substrate underlying simple horizontal saccadic eye movements was investigated using echoplanar functional magnetic resonance imaging (fMRI) in young healthy volunteers. Echoplanar imaging with signal targeting and alternating radiofrequency (EPISTAR), a novel perfusion technique, measured signal intensity changes in one to four contiguous 10-mm slices centered to include both striate cortex and putative frontal eye fields during horizontal saccade and fixation conditions. Subtraction images of self paced visually guided saccadic versus fixation conditions showed bilateral marked and statistically significant localized signal increases in the precentral region (Brodmann areas 4, 6) and peristriate cortex (areas 17, 18, 19) and qualitative increases in the superior medial frontal region, as identified by a Talairach Tournoux generalized template in the brain slices that were scanned. Additional parietal activation occurred during a target-guided saccade task. Our data (i) support the localization of the human FEF, as identified by simple, nonexploratory saccadic eye movements, in the precentral motor strip and premotor cortex, (ii) show individual variability in the exact anatomical location of saccade-related activations, and (iii) confirm that the EPISTAR technique can demonstrate localized signal increases during a behavioral task. PMID- 9345476 TI - Spatial activation maps--bioelectric signals related to 3D recorded movements. AB - The analysis of multidimensional neurophysiological data poses difficulties for the scientist in understanding the often complex inherent data relations. This is particularly the case when neuromuscular cell discharge parameters are to be related to body segment movement characteristics in freely behaving animals or humans. The understanding of such data is greatly simplified if the recorded data can be visualized in an adequate way, and relevant data relations are thus highlighted. This report describes a rationale for display and qualitative analyses of muscle discharge patterns in relation to three-dimensionally recorded limb or body movements. The method is illustrated by creating spatial activation plots and spatial activation maps of the electromyographic activity in the first interosseal muscle in the human hand, but this rationale for data presentation can be applied to most excitable tissues that are activated in relation to body or limb movements. PMID- 9345477 TI - Isolating the mnemonic component in spatial delayed response: a controlled PET 15O-labeled water regional cerebral blood flow study in normal humans. AB - In this study we attempted to elucidate the neural network involved in maintaining spatial information over short delays by means of the PET 15O-labeled water method for measuring regional cerebral blood flow. To isolate the mnemonic component of delayed response processing we designed a control task isomorphic to the experimental task (in which the subject remembered the location of four targets in an array over a 7 s delay) and controlled for spatial encoding. Fourteen normal subjects participated in the study. Regional cerebral blood flow (rCBF) data were analyzed using statistical parametric mapping, a data-driven approach which canvasses the whole brain for significantly activated pixels in the experimental vis-a-vis the control task, and a hypothesis driven region of interest approach involving comparisons of control and experimental conditions using normalized rCBF data. We found convergent evidence for a spatially distinct but functionally related network in which dorsolateral prefrontal cortex and extrastriate occipital cortex were activated by the experimental task vis-a-vis the control task, as well as evidence for superior parietal and supplementary motor area cortical activation. These findings suggest that anterior components of the network may be involved in mnemonic rehearsal functions, while posterior components may store critical perceptual attributes of the memoranda. PMID- 9345478 TI - PET evidence for an amodal verbal working memory system. AB - Current models of verbal working memory assume that modality-specific representations are translated into phonological representations before entering the working memory system. We report an experiment that tests this assumption. Positron emission tomography measures were taken while subjects performed a verbal working memory task. Stimuli were presented either visually or aurally, and a visual or auditory search tasks, respectively, was used as a control. Results revealed an almost complete overlap between the active memory areas regardless of input modality. These areas included dorsolateral frontal, Broca's area, SMA, and premotor cortex in the left hemisphere; bilateral superior and posterior parietal cortices and anterior cingulate; and right cerebellum. These results correspond well with previous research and suggest that verbal working memory is modality independent and is mediated by a circuit involving frontal, parietal, and cerebellar mechanisms. PMID- 9345479 TI - A new technique for quantitative imaging of cerebrovascular reserve capacity using a double injection method with N-isopropyl-p-[123I]iodoamphetamine. AB - Estimation of local cerebrovascular reserve capacity is an important aspect of the management of patients with cerebrovascular occlusive disease. Regional normal values of cerebrovascular reserve capacity have not yet been established. Recently, Mori et al. (1994) introduced a method to quantitatively measure cerebral blood flow twice in 30 min after a double injection of N-isopropyl-p [123I]iodoamphetamine using a background subtraction method. We utilized this double injection method in tandem with acetazolamide (20 mg/kg, intravenous injection) in 10 normal volunteers to produce a map of quantitative cerebrovascular reserve capacity using single photon emission computed tomography. The normal regional cerebrovascular reserve capacity was: frontal lobe, 51.0 +/- 19.8%; temporal lobe, 53.6 +/- 12.4%; parietal lobe, 69.3 +/- 31.8%; basal ganglia, 85.1 +/- 42.7%; cerebellum, 49.6 +/- 17.9% (mean +/- SD). One-way analysis of variance showed that the cerebrovascular reserve capacity did not differ significantly among structures. This new imaging technique and normal values of regional cerebrovascular reserve capacity may provide information about the pathophysiology and surgical indication in patient with occlusive cerebrovascular disease. PMID- 9345480 TI - Visualizing cortical activation during mental calculation with functional MRI. AB - Cortical activation during arithmetic calculation (silent subtraction by sevens) was compared to that observed during a control condition for which subjects were required to count forward by ones. Nine normal subjects underwent 1.5-T functional magnetic resonance imaging while performing these tasks. All subjects showed bilateral premotor, posterior parietal, and prefrontal cortex activation during serial calculation. There was a large degree of individual variation in activation outside of these areas. These results confirm the role of posterior parietal cortex in arithmetic calculation and implicate other regions, including prefrontal cortex. PMID- 9345481 TI - Brain activity related to the perception of illusory contours. AB - We have addressed the question of whether the brain's capacity to resolve an ambiguous retinal image depends upon the activity of early visual areas or whether it involves the investment of the received image with higher order cognitive hypotheses. To resolve the issue, we have used the technique of positron emission tomography to detect increases in regional cerebral blood flow (rCBF) in the brains of humans while they perceive the simple figures described by Schumann (1900) and by Kanizsa (1979). These figures produce striking percepts of surfaces or contours variously described as illusory, subjective, cognitive, or anomalous because they depend upon the brain's ability to complete the figures. If such completion is due to higher order cognitive processes or a combination of higher order and early areas, then, one might expect areas of increased rCBF outside the occipital lobe when subjects perceive these figures. However, if completion is mediated entirely by early visual areas, then the increases in rCBF will be restricted to these regions. Our results show that the perception of subjective contours is associated with significant activity in early visual areas only, particularly in area V2, leading us to conclude that the occipital cortex can contribute to the perception of these stimuli without higher order cognitive influence specific to the completion task. PMID- 9345482 TI - MRI-guided flumazenil- and FDG-PET in temporal lobe epilepsy. AB - In temporal lobe epilepsy (TLE) patients without lesions, major hippocampal sclerosis, or atrophy on magnetic resonance imaging (MRI), the localizing power of [11C]flumazenil (FMZ) and 2-[18F]fluoro-2-deoxy-D-glucose (FDG) was compared using high-resolution positron emission tomography (PET) studies and individually coregistered MRI scans. Following complete clinical, neuropsychological, and electrophysiological evaluation, benzodiazepine receptor density was assessed using the FMZ equilibrium method. Thirty minutes later, interictal FDG-PET was performed under resting conditions. PET images were matched to three dimensionally coregistered, T1-weighted MRI. Each temporal lobe (TL) was divided into 12 volumes of interest. The regional FMZ data were normalized with respect to average cortical values. For each patient the right-left asymmetries of rCMRGlc and normalized FMZ data were calculated. In 7 to 10 patients, mesial TL structures showed reduced FMZ binding, with a decrease by at least 10% in the affected TL. Reductions of 10% or more of rCMRGlc usually were more widespread than FMZ reductions and often involved lateral temporal cortex. The regions of most pronounced disturbances are not necessarily identical in both methods. Three patients had a complex correspondence of lateralization with PET, neuropsychological, and EEG data. In 4 patients, lateralization was less clear from EEG or neuropsychological results but was still consistent with lateralization by PET. In 3 of 10 patients, however, major discrepancies were found. These data suggest that the combination of neuropsychological testing, EEG, and MRI-guided FMZ- and FDG-PET will help to select patients with clearly defined epileptogenic foci especially in mesial TLE. Even in cases without MRI lesions, TL epileptic foci can be lateralized with consistency across the methods; FMZ-PET shows the pathologic focus more circumscribed than FDG-PET. PMID- 9345483 TI - Brain activation induced by estimation of duration: a PET study. AB - Duration information about a visual stimulus requires processing as do other visual features such as size or intensity. Using positron emission tomography, iterative H215O infusions, and statistical parametric mapping, we investigated the neural correlates of time processing. Nine normal subjects underwent six serial rCBF. Three tasks were studied: (a) A temporal generalization task (D task) in which the subjects had to judge (by pressing one of two keys) whether the duration of the illumination of a green LED was equal to or different from that of a previously presented standard; (b) An intensity generalization task (I task) in which the judgment concerned the intensity of the LED; and (c) A control task (C task) in which the subjects had to press one of the two keys at random in response to LED illumination. A significant increase in rCBF during the D task, compared to that during the C task, was observed in right prefontal cortex, right inferior parietal lobule, anterior cingulate cortex, vermis, and a region corresponding to the left fusiform gyrus. A significant increase in rCBF during the I task, compared to that during the C task, was observed in right prefontal cortex, right inferior parietal lobule, right extrastriate cortex, anterior cingulate cortex, left inferior parietal lobule, vermis, and two symmetrical regions corresponding to the fusiform gyri. No significant activation was observed in the D task when compared to that in the I task. We propose that these cortical maps are best explained by the recruitment of visual attention and memory structures, which play a major role in prospective time judgements as indicated by behavioral studies. The data also suggest that the temporal dimension of a visual stimulus is processed in the same areas as other visual attributes. PMID- 9345484 TI - Adult age differences in regional cerebral blood flow during visual world identification: evidence from H215O PET. AB - We used H215O PET to investigate adult age differences in regional cerebral blood flow (rCBF) during the performance of a visual word identification task. The study participants were 20 healthy, right-handed men: 10 young adults between 18 and 27 years of age, and 10 older adults between 63 and 75 years of age. The word identification task comprised six blocks of test trials representing four task conditions; subjects responded manually. The task conditions varied with regard to whether semantic retrieval was required (e.g., word/nonword discrimination vs simple response to each stimulus) and with regard to the difficulty of visual encoding (e.g., words presented normally vs words with asterisks inserted between adjacent letters). Each subject performed all six trial blocks, concurrently with each of six H215O PET scans. Analyses of quantitative CBF data obtained from the arterial time-activity curve demonstrated a significant age-related decline in global CBF rate. Analyses of the changes in rCBF between task conditions indicated that retrieval of semantic information sufficient to distinguish words from nonwords is mediated by a ventral occipitotemporal cortical pathway. Specific areas within this pathway were also associated with visual encoding processes. Several rCBF activations were significantly greater for young adults than for older adults, indicating an age-related decline in processing efficiency within this ventral occipitotemporal pathway. Although the performance data demonstrated a greater age-related slowing for visual encoding than for semantic retrieval, these age-related performance changes were not associated with corresponding changes in rCBF activation. PMID- 9345485 TI - Spatial pattern analysis of functional brain images using partial least squares. AB - This paper introduces a new tool for functional neuroimage analysis: partial least squares (PLS). It is unique as a multivariate method in its choice of emphasis for analysis, that being the covariance between brain images and exogenous blocks representing either the experiment design or some behavioral measure. What emerges are spatial patterns of brain activity that represent the optimal association between the images and either of the blocks. This process differs substantially from other multivariate methods in that rather than attempting to predict the individual values of the image pixels, PLS attempts to explain the relation between image pixels and task or behavior. Data from a face encoding and recognition PET rCBF study are used to illustrate two types of PLS analysis: an activation analysis of task with images and a brain-behavior analysis. The commonalities across the two analyses are suggestive of a general face memory network differentially engaged during encoding and recognition. PLS thus serves as an important extension by extracting new information from imaging data that is not accessible through other currently used univariate and multivariate image analysis tools. PMID- 9345486 TI - A new in vivo method for quantitative analysis of stroke lesions using diffusion weighted magnetic resonance microscopy. AB - Using three-dimensional diffusion-weighted MR microscopy and a rat model of focal cerebral ischemia, we evaluated the statistical characteristics of two parameters: absolute stroke volumes and change in stroke volumes over 6 h of middle cerebral artery (MCA) occlusion. In all rats, the absolute stroke volumes increased linearly over the 6-h MCA occlusion time period. On average, stroke volume growth rate was 2.1 +/- 0.5%/h. Sample size power analysis of our data demonstrated that to demonstrate a 10% reduction of the 6-h volumes, sample size per group would require 29 animals (these calculations are based on alpha = 0.05, beta = 0.20 using normal approximation). A similar 30% reduction of stroke volume at 6 h poststroke (from approximately equal to 200 to 140 mm3) would, in our "slope model," translate into a reduction of stroke growth rate from the normal + 11.25 mm3/h (150 to 200 mm3 over 4 h) to 7 mm3/h (150 to 178 mm3 over 4 h); power analysis in this case demonstrated that sample size is reduced to 15 animals per group (these calculations are based on alpha = 0.05, beta = 0.20 using normal approximation). We conclude that from a statistical standpoint our study demonstrates that stroke growth rate might be a more suitable parameter for evaluating the effect of treatment in both clinical and experimental stroke trials. PMID- 9345488 TI - Normalizing counts and cerebral blood flow intensity in functional imaging studies of the human brain. AB - Image intensity normalization is frequently applied to eliminate or adjust for subject or injection global blood flow (gCBF) and other sources of nuisance variation. Normalization has several other positive effects on the analysis of PET images. However, the choice of an intensity normalization technique affects the statistical and psychometric properties of the image data. We compared three normalization procedures, the ratio approach (regional (r)CBF/gCBF), histogram equalization, and ANCOVA, on both PET count and flow data sets. The ratio method presents the proportional increase of regions, the histogram equalization method offers the relative ranking of intensities over the image, and the ANCOVA method provides statistical deviations from an expected linear model of regional values from the subject's gCBF. The original study used 33 normal subjects in a standard subtraction paradigm. The normalization methods were evaluated on their ability to remove extraneous error variation, induce homogeneity of intersubject variation, and remove unwanted dependencies. In general, the normalization modified the subtraction image more than the individual condition images. All three methods worked well at removing the dependency of rCBF on gCBF in count and flow images. For count data, the three methods also reduced the amount of error variation equally well, improving the signal to noise ratio. For flow data, the histogram equalization and ratio methods worked best at reducing statistical error. All three methods dramatically stabilized the variance over the image. PMID- 9345487 TI - A multivariate analysis of evoked responses in EEG and MEG data. AB - This paper presents a multivariate analysis of evoked responses and their spatiotemporal dynamics as measured with electro- or magnetoencephalography. This analysis uses standard techniques (ManCova) to make possible statistical inference about differential responses, after the data have been transformed using singular value decomposition. The generality of this approach is limited only by the assumptions implicit in the general linear model and can range from simple analyses like Hotelling's T2 test (in comparing evoked responses among different conditions) to complex analyses of a multivariate regression type (e.g., characterizing the response components associated with a behavioral or psychophysical parameter). To illustrate the technique we have characterized time dependent changes (both within and between trials) in magnetic fields, evoked by self-paced movements. Our illustrative analysis showed that movement-evoked components were less prone to adaptation than premovement components, suggesting that functionally distinct (preparatory and early executive) biomagnetic signals show differential adaptation. PMID- 9345489 TI - Individual functional anatomy of verb generation. AB - Examination of the individual functional anatomy of language is of particular interest in clinical neurology to explain the variability of aphasic symptoms after focal lesions and to avoid damage of language-related brain areas by surgery. For a silent verb generation task, we examined whether activation PET with 3D data acquisition, multiple replication of conditions, and coregistration with MRI provides results that are consistent and reproducible enough to be useful clinically. Visual analysis was performed on PET-MRI fusion images, including renderings of the brain surface. Quantitative analysis was based on volumes of interest. In seven right-handed normals, activation of the triangular part of the left inferior frontal cortex [Brodman area (BA) 45] was the most significant finding that was present in each subject. Two subjects showed minor anatomical variants of the ascending or horizontal ramus of the sylvian fissure that were associated with the least activation of BA 45. In the left hemisphere the other frontal gyri, the superior temporal and posterior part of the middle temporal gyrus, and the paracingulate gyrus were also significantly activated. There was significant bilateral cerebellar activation, but it was significantly more intense on the right than on the left side. The consistency and high interindividual reproducibility of these findings suggest that this technique may be useful for clinical assessment of language-related areas. PMID- 9345490 TI - Focal cortical blood flow activation is regulated by intrinsic cortical cholinergic neurons. AB - We evaluated the cholinergic mechanism underlying focal cortical vascular response to neuronal activation, using positron emission tomography for use on animals to measure cerebral blood flow and glucose metabolism activation upon vibrotactile stimulation in cats. Bromopyruvate, which blocks acetylcholine synthesis through inhibition of the production of acetyl CoA, was injected into the cerebral cortex and basal forebrain as well as the sphenopalatine ganglion, all of which have been confirmed to supply cholinergic terminals to the cerebral cortex. Although glucose metabolism was preserved, indicating that the neuronal activities were enhanced, cerebral blood flow increase during cortical neuronal activation was abolished by bromopyruvate injection into only the cerebral cortex and not other cholinergic systems. We conclude that the cholinergic intrinsic neurons control the focal cerebral blood flow increase in response to neuronal activation. PMID- 9345491 TI - The evolution of optical signals in human and rodent cortex. AB - The time course of optical intrinsic signals was examined in order to characterize the evolution of response in human and rodent cortex. Both subtraction/ratio and principal component analyses were used to construct time course curves. The time course began at a prestimulus baseline, responded with a finite delay, overcompensated, reduced to a maintenance level, and then disappeared. The magnitude, spatial involvement, and principal components demonstrated similar time-course curves both in human and in rodent. For acute stimuli, peak response was reached between 2 and 3 s and returned to baseline by 6 s poststimulation. The shape of the time-course curve is consistent with the need to satisfy neuronal demand and the contributions of vascular smooth muscle properties to the response behavior. The temporal delays and nonlinear phenomena observed in the time-course curves are consistent with a hydraulic model of neurovascular supply/demand behavior. PMID- 9345492 TI - Is multivariate analysis of PET data more revealing than the univariate approach? Evidence from a study of episodic memory retrieval. AB - In a functional imaging study of cued paired associate retrieval, in which the strength of association between pair members was systematically varied, we predicted increased right frontal activity as a function of weakening semantic linkage. An initial univariate analysis found the opposite effect, with greater right frontal activity during recall of strongly linked paired associates. This unexpected result led us to perform a multivariate analysis of covariance (MANCOVA), an approach which proved more informative. This analysis showed that the most significant source of task-related variance was accounted for by a nonlinear relationship not predicted by the prior hypothesis and not revealed by the standard univariate approach. This application of the MANCOVA supports the assertion that multivariate analysis can provide an important adjunct to univariate approaches like statistical parametric mapping (SPM). New perspectives engendered by the MANCOVA still allow for statistical inference but are not constrained by explicit hypotheses about specific task-dependent effects. PMID- 9345493 TI - Functional magnetic resonance imaging of human visual cortex during face matching: a comparison with positron emission tomography. AB - Cortical areas associated with the perception of faces were identified using functional magnetic resonance imaging (fMRI). T2*-weighted gradient echo, echo planar MR images were obtained using a modified 1.5-T GE Signa MRI. In all nine subjects studied, performance of a face-matching task was associated with a region of significantly increased MR signal in the ventral occipitotemporal cortex, extending from the inferior occipital sulcus to the lateral occipitotemporal sulcus and fusiform gyrus. Smaller and more variable signal increases were found in dorsolateral occipitoparietal cortex near the intraparietal sulcus. Signal decreases were found in the angular gyrus and posterior cingulate cortex. Single-subject fMRI analyses revealed discrete areas of activation with well-defined borders. Group analyses of spatially smoothed fMRI data produced results that replicated most aspects of previous studies of face processing using positron emission tomography (PET). These results show that PET and fMRI identify functional areas with similar anatomical locations. In addition, fMRI reveals interindividual variation in the anatomical location of higher-level processing areas with greater anatomical precision. PMID- 9345494 TI - Functional magnetic resonance image analysis of a large-scale neurocognitive network. AB - Many "higher-order" mental functions are subserved by large-scale neurocognitive networks comprising several spatially distributed and functionally specialized brain regions. We here report statistical and graphical methods of functional magnetic resonance imaging data analysis which can be used to elucidate the functional relationships (i.e., connectivity and distance) between elements of a neurocognitive network in a single subject. Data were acquired from a normal right-handed volunteer during periodic performance of a task which demanded visual and semantic processing of words and subvocalization of a decision about the meaning of each word. Major regional foci of activation were identified (by sinusoidal regression modeling and spatiotemporal randomization tests) in left extrastriate cortex, angular gyrus, supramarginal gyrus, superior and middle temporal gyri, lateral premotor cortex, and Broca's area. Principal component (PC) analysis was initially undertaken by singular value decomposition (SVD) of the "raw" time series observed at 170 activated voxels. This revealed a large functional distance (negative connectivity) between visual processing systems and all other brain regions in the space of the first PC. SVD of a matrix of fitted time series, and a matrix of six sinusoidal regression parameters estimated at each activated voxel, were developed as less noisy (more informative) alternatives to SVD of the "raw" data. Canonical variate analysis of denoised data was then used to clarify functional relationships between the major regional foci. Visual input analysis systems (extrastriate cortex and angular gyrus) were colocalized in the space of the first canonical variate (CV) and significantly separated from all other brain regions. Semantic analysis systems (supramarginal and temporal gyri) were colocalized and significantly separated in the space of the second CV from the subvocal output system (Broca's area). These results are provisionally interpreted in terms of underlying hemodynamic events and cognitive psychological theory. PMID- 9345495 TI - Reproducibility of PET activation studies: lessons from a multi-center European experiment. EU concerted action on functional imaging. AB - PET activation studies are performed widely to study human brain function. The question of reproducibility, reliability, and comparability of the results of such experiments has never been addressed on a large scale. Recently, 12 European PET centers performed the same cognitive activation experiment in a European Union funded concerted action. The experiment involved a standardized and validated cross-lingual experimental and control task involving verbal fluency. Each center contributed at least 6 subjects. In total there were 77 subjects and 247 scans in each of the two conditions, giving 494 scans in total. We have analyzed each center's dataset and pooled datasets using statistical parametric mapping. We present results that address the consistency of these analyses, discuss the factors that influence their sensitivity, and comment on a number of related methodological issues. We used a MANOVA to test for center, condition, and centre by condition effects and found a strong condition and center effect and weaker interactions. The main effect determining reproducibility was the overall sensitivity of the experiment, to which the scanner and number of scans contribute in a major way, with a marked advantage for 3D scanners and a large field of view. An important conclusion is that data from different centers can be pooled to improve the reliability of results, which is of particular importance for studies in patients with rare conditions. PMID- 9345496 TI - Functional activation during an auditory comprehension task in patients with temporal lobe lesions. AB - Functional magnetic resonance imaging (fMRI) was used to map regional brain activation during an auditory comprehension task in two normal controls and two patients with left temporal lobe lesions. Activity in the superior temporal and angular gyrus regions was detected in all normal subjects. In the patients, the spatial distribution of activation ipsilateral to the lesions differed from the pattern observed in contralateral cortex or in control subjects. These studies highlight the potential of fMRI for mapping abnormal functional anatomy in the human brain. PMID- 9345498 TI - Functional connectivity in auditory-verbal short-term memory in Alzheimer's disease. AB - Alzheimer's disease (AD) patients show normal patterns of regional cerebral blood flow during performance of subspan verbal memory tasks, but appear to allocate new brain regions when performing supraspan recall tasks. To understand the relationships among these interconnected brain regions, their functional connectivity was considered using principal components analysis (PCA). AD patients and controls were scanned in four conditions: visual fixation only and visual fixation with one-word recall, three-word recall, and eight-word recall. When applied to the data from the three recall task conditions (i.e., eliminating fixation), PCA revealed a significant first component accounting for 75.8% of the variance in the control data and 66.8% of the variance in the patient data. The one-word recall condition had a high positive component score, and the eight-word condition a high negative score. The correlations of individual voxels on this factor (i.e., spatial modes) were very similar between the patients and the controls. The overall similarity in the results of the PCAs between the patients and the controls suggests that they have similar functional connectivity. This would suggest that, at least in the early course of the dementia, the functional CNS organization of verbal memory systems remains normal. PMID- 9345497 TI - Nonlinear regression in parametric activation studies. AB - Parametric study designs can reveal information about the relationship between a study parameter (e.g., word presentation rate) and regional cerebral blood flow (rCBF) in functional imaging. The brain's responses in relation to study parameters might be nonlinear, therefore the (linear) correlation coefficient as often used in the analysis of parametric studies might not be a proper characterization. We present a noninteractive method, which fits nonlinear functions of stimulus or task parameters to rCBF responses, using second-order polynomial expansions. This technique is implemented in the context of the general linear model and statistical parametric mapping. We also consider the usefulness of statistical inferences, based on F fields, about similarities and differences of these nonlinear responses in different groups. This approach is illustrated with a 12-run H215O PET activation study using an auditory paradigm of increasing word presentation rates. A patient who had recovered from severe aphasia and a normal control were studied. We demonstrate the ability of this new technique to identify brain regions where rCBF is closely related to increasing word presentation rate in both subjects without constraining the nature of this relationship and where these nonlinear responses differ. PMID- 9345499 TI - Imaging motor-to-sensory discharges in the human brain: an experimental tool for the assessment of functional connectivity. AB - We present a new approach to studying functional connectivity in the human brain. This approach is based on the observation that when we engage in motor activity, a discharge corollary to the motor command is sent from motor to sensory structures. Thus, as long as movement-related sensory input is either prevented or masked, modulation of neuronal activity in sensory structures would indicate the presence of functional connectivity between the motor and the sensory regions. Using positron emission tomography, such a central interaction between motor and sensory regions can be assessed by measuring regional changes in cerebral blood flow (CBF) in sensory regions. In this paper, we describe the experimental design and the results of two studies of corollary discharges, namely those generated during eye movements and speech. In these studies, a graded approach was used to establish the relationship between the number of eye movements or utterances and CBF in visual or auditory regions, respectively. Significant covariations between the number of movements and CBF in sensory regions were found, thus indicating the presence of functional connectivity between motor and sensory regions. In addition, interregional CBF covariations were computed and the effect of removing the intersubject variance on these covariations was evaluated. The corollary-discharge-based approach to studying functional connectivity is discussed in the context of more traditional computational approaches to network analysis in functional brain imaging. PMID- 9345500 TI - Functional basis of memory impairment in multiple sclerosis: a[18F]FDG PET study. AB - Structural neuroimaging has been used to correlate lesional patterns with the cognitive profile of patients with multiple sclerosis (MS), especially for "frontal" dysfunction. However, a clear-cut anatomical explanation has yet to be found for the long-term memory deficit which is a hallmark of MS cognitive impairment. We have used PET to measure regional cerebral glucose metabolism (rCMRglc) in a group of 15 MS patients with involvement of verbal and/or spatial long-term memory. These patients were compared with 10 normal controls and 13 MS patients unimpaired on all neuropsychological tests. Relative to the controls, MS patients with memory deficits showed a significant bilateral reduction of rCMRglc in the hippocampus, cingulate gyrus, thalamus, associative occipital cortex, and cerebellum. Direct comparisons between patients with memory deficits and the group of unimpaired MS patients showed a metabolic reduction in the left thalamus and in both hippocampi. Seven of the memory-impaired patients also had neuropsychological signs of frontal dysfunction. These patients were compared with patients who had isolated memory deficit. Here we observed a further metabolic reduction in a number of brain regions including bilateral prefrontal cortex, inferior parietal cortex, and basal ganglia. Our findings indicate that hypometabolism of thalamic and deep cortical gray structures of the temporal lobe is associated with episodic memory dysfunction in MS. On the other hand, pathological performance on tests designed to assess frontal functions was associated with widespread reduction of glucose metabolism. PMID- 9345501 TI - The trouble with cognitive subtraction. AB - In this paper we present a critique of pure insertion. Pure insertion represents an implicit assumption behind many (but not all) studies that employ cognitive subtraction. The main contention is that pure insertion is not valid in relation to the neuronal instantiation of cognitive processes. Pure insertion asserts that there are no interactions among the cognitive components of a task. It is possible to evaluate and refute this assumption by testing explicitly for interactions using factorial experimental designs. It is proposed that factorial designs are more powerful than subtraction designs in characterizing cognitive neuroanatomy, precisely because they allow for interactions and eschew notions like pure insertion. In particular we suggest that the effect of a cognitive component (i.e., an effect that is independent of other components) is best captured by the main (activation) effect of that component and that the integration among components (i.e., the expression of one cognitive process in the context of another) can be assessed with the interaction terms. In this framework a complete characterization of cognitive neuroanatomy includes both regionally specific activations and regionally specific interactions. To illustrate our point we have used a factorial experimental design to show that inferotemporal activations, due to object recognition, are profoundly modulated by phonological retrieval of the object's name. This interaction implicates the inferotemporal regions in phonological retrieval, during object naming, despite the fact that phonological retrieval does not, by itself, activate this region. PMID- 9345502 TI - Brain representation of active and passive movements. AB - During active and passive (driven by a torque motor) flexion and extension of the right elbow, regional cerebral blood flow (rCBF) was measured in six healthy, male volunteers using positron emission tomography and the standard H2(15)O injection technique. During active as well as during passive movements of the right elbow there were strong increases in rCBF, identical in location, amount, and extent in the contralateral sensorimotor cortex. There were activations during both conditions in the supplementary motor area (stronger and more inferior in the active condition) and inferior parietal cortex (on the convexity during active movements and in the depth of the central sulcus during passive movements). During active movements only, activations of the basal ganglia and the cingulate gyrus were found. Brain activations during motor tasks are largely related to the processing of afferent information. PMID- 9345503 TI - Time-varying activation of different cytoarchitectonic areas of the human SI cortex after tibial nerve stimulation. AB - We followed cortical activation in eight healthy adults after electric stimulation of the left tibial nerve at the ankle. The recordings were made noninvasively with a whole-scalp neuromagnetometer. The first cortical activation peaked in different subjects at 37-45 ms in the foot area of the right (contralateral) primary somato-sensory (SI) cortex, with mean source current orientation perpendicular to the longitudinal fissure. The current orientation changed within the next 5 ms counterclockwise in all subjects, with a mean rotation of 64 degrees. A two-dipole time-varying model, with two dipoles differing by 28-119 degrees in orientation but less than 1 cm in location in the right SI cortex, explained the signal pattern satisfactorily during the first 100 ms. We suggest that the observed field patterns reflect sequential activation of different cytoarchitectonic areas in the foot SI cortex and imply considerable differences in the structural organization between the foot and the hand SI cortices. The initial activation is considered to take place in area 3b facing the interhemispheric fissure, and the later source, due to the systematic rotations of the field patterns, is assumed to reflect activation of area 5 in the anterior wall of the marginal ramus of the cingulate sulcus. PMID- 9345504 TI - A stereotaxic template atlas of the macaque brain for digital imaging and quantitative neuroanatomy. AB - A stereotaxic brain atlas of the longtailed macaque (Macaca fascicularis) is presented in a format suitable for use as a template atlas of the macaque brain. It includes most of the brain segmented to show the boundaries of landmark structures such that every point in the brain can be represented by a unique set of coordinates in three-dimensional space and ascribed unambiguously to one and only one primary structure. More than 400 structures are represented, including 360 volumetric structures, which constitute the substance of the brain, and 50 superficial features. To facilitate use with ventriculography, magnetic resonance imaging, and other noninvasive imaging techniques, the stereotaxic space is referenced to internal landmarks, viz., the anterior commissure and posterior commissure; the center of the anterior commissure at the midline is the origin of the stereotaxic axes. Reference of stereotaxis to this bicommissural space facilitates structural comparison with human brain atlases, which are commonly referenced to the biocommissural line. It also facilitates comparison of brains of different nonhuman primate species by providing a template brain against which to compare size and internal variability. Thirty-three coronal sections at 1-mm intervals from the spinomedullary junction to the rostral extreme of the caudate nucleus show most structures of the hindbrain, midbrain, and subcortical forebrain. Separately, four side views and 16 coronal sections show cortical structures. Structures are represented by outlines of their boundaries and labeled according to NeuroNames, a systematic English nomenclature of human and nonhuman primate neuroanatomy. Abbreviations are based on a protocol designed to facilitate cross-species comparisons. Instructions are provided for: (1) locating sites from the Template Atlas in the conventional stereotaxic space of an experimental animal, (2) locating sites identified by conventional stereotaxis in the Template Atlas, and (3) using the Template Atlas to collate, compare, and display image information (e.g., labeled cells, recording sites, stimulation sites, lesions) from multiple animals. PMID- 9345505 TI - Simplified reference tissue model for PET receptor studies. AB - The reference tissue model allows for quantification of receptor kinetics without measuring the arterial input function, thus avoiding arterial cannulation and time-consuming metabolite measurements. The model contains four parameters, of which the binding potential (BP) is the parameter of interest. Although BP is robust, convergence rates are slow and the other parameters can have large standard errors. To overcome this problem, a simplified reference tissue containing only three parameters was developed. This new three-parameter model was compared with the previous four-parameter model using a variety of PET studies: [11C]SCH 23390 (D1 receptor) and [11C]raclopride (D2 receptor) in humans, and [11C]SCH 23390, [11C]raclopride and [11C]RTI-121 (dopamine transporter) in rats. The BP values obtained from both models were essentially the same for all cases. In addition, the three-parameter model was insensitive to starting values, produced stable results for the other parameters (small standard errors), and converged rapidly. In conclusion, for the ligands tested the three parameter model is a better choice, combining increased convergence rate with increased stability. PMID- 9345506 TI - Auditory selective attention: an fMRI investigation. AB - In the present experiment, 25 adult subjects discriminated speech tokens ([ba]/[da]) or made pitch judgments on tone stimuli (rising/falling) under both binaural and dichotic listening conditions. We observed that when listeners performed tasks under the dichotic conditions, during which greater demands are made on auditory selective attention, activation within the posterior (parietal) attention system and at primary processing sites in the superior temporal and inferior frontal regions was increased. The cingulate gyrus within the anterior attention system was not influenced by this manipulation. Hemispheric differences between speech and nonspeech tasks were also observed, both at Broca's Area within the inferior frontal gyrus and in the middle temporal gyrus. PMID- 9345507 TI - Functional magnetic resonance imaging of motor activation in the human cervical spinal cord. AB - Functional magnetic resonance imaging (fMRI) at 1.5 T of a 30-mm segment of the human spinal cord, centered at the seventh cervical cord segment, showed mean blood-oxygenation-dependent contrast changes in image intensity of 4.8% associated with a unilateral hand-closing task in normal human volunteers. The observed locale of activation in the ipsilateral intermediate and ventral gray matter of the cervical cord contains motoneurons, corticospinal axonal terminations from the hand area of the brain motor cortex, and capillaries supplying the spinal neurons. This noninvasive observation of focal activation within the human spinal cord is consistent with neuronal cooperation over more than one cord segment and suggests that fMRI of the human central nervous system may have wider clinical applications outside of the brain. PMID- 9345508 TI - Cerebral vasomotion: a 0.1-Hz oscillation in reflected light imaging of neural activity. AB - Imaging of scattered and reflected light from the surface of neural structures can reveal the functional architecture within large populations of neurons. These techniques exploit, as one of the principal signal sources, reflectance changes produced by local variation in blood volume and oxygen saturation related to neural activity. We found that a major source of variability in the captured light signal is a pervasive low-frequency (0.1-Hz) oscillation which apparently results from regional cerebral blood flow. This signal is present in brain parenchyma as well as the microvasculature and exhibits many characteristics of the low-frequency "vasomotion" signals observed in peripheral microcirculation. Concurrent measurements in brain with a laser Doppler flow meter contained an almost identical low-frequency signal. The presence of the 0.1-Hz oscillation in the cerebral microcirculation could underlie a portion of the previously described characteristics reported in reflected-light imaging studies. The prevalence of the oscillatory phenomena in the brain raises substantial temporal sampling issues for optical imaging and for other visualization techniques which depend on changes in regional cerebral blood dynamics, such as functional magnetic resonance imaging. PMID- 9345509 TI - Neural activation during covert processing of positive emotional facial expressions. AB - Lesion studies indicate distinct neural systems for recognition of facial identity and emotion. Split-brain experiments also suggest that emotional evaluation of a stimulus can occur without conscious identification. The present study tested a hypothesis of a differential neural response, independent of explicit conscious mediation, to emotional compared to nonemotional faces. The experimental paradigm involved holding in mind an image of a face across a 45-s delay while regional cerebral blood flow was measured using positron emission tomography. Prior to the delay, a single face was presented with an explicit instruction to match it to one of two faces, photographed at different angles from the target face, presented at the end of the delay. Repeated blood flow measures were obtained while subjects held happy or neutral faces in mind or during a neutral control fixation condition without initial face presentation. The representation of emotional faces over a delay period, compared to either the nonemotional or the fixation condition, was associated with significant activation in the left ventral prefrontal cortex, the left anterior cingulate cortex, and the right fusiform gyrus. The findings support our hypothesis of a differential neural response to facial emotion, independent of conscious mediation, in regions implicated in the processing of faces and of emotions. PMID- 9345510 TI - Quantitative comparison of functional magnetic resonance imaging with positron emission tomography using a force-related paradigm. AB - The intention of our study was to compare functional magnetic resonance imaging (fMRI) with positron emission tomography (PET). We used the same force-related motor paradigm for both techniques, which allows for quantification of stimulus intensity. Regional cerebral blood flow (rCBF) was determined with PET in six male subjects (age 30 +/- 3) using the slow bolus injection technique and oxygen 15-labeled water. Scans were collected during six different conditions: at rest and during repetitive Morse key press at 1 Hz, with the right index finger at a range of different forces. In a second series of experiments fMRI data were acquired under similar conditions in six volunteers in a single slice parallel to and 51 +/- 3 mm dorsal to the anterior and posterior commissure (AC-PC). A conventional 1.5-T clinical magnetic resonance (MR) system and the FLASH technique were used. The data obtained in both series of experiments were subjected to the same statistical analyses. Statistical parametric maps (SPM) were generated by two different approaches: a correlation between peak force and rCBF or fMRI signal and using a categorical comparison of force exerted with rest. SPMs were coregistered with anatomical MR images. PET and fMRI measurements demonstrated activation in the primary motor cortex (M1) and posterior supplementary motor cortex in all subjects. Correlation analysis demonstrated foci in the M1 in four subjects with PET and in only one subject with fMRI. Locations of activation peaks differed by 2 to 8 mm between imaging methods. The relationship between fMRI signal or rCBF and peak force was logarithmic. The maximum increase in fMRI signal was 5.0% +/- 0.9 at 60% of the maximum voluntary contraction while the corresponding increase in rCBF was 13.7% +/- 1.2. The ratio of percentage rCBF change to percentage fMRI signal change was very similar across all force levels. The high degree of correspondence between PET and fMRI data provides good cross-validation for the two techniques. PMID- 9345511 TI - The role of the thalamus in "top down" modulation of attention to sound. AB - By correlating rCBF with rate of presentation of tones we used PET to identify brain regions where auditory signals elicited a transient neural response. In one condition volunteers were asked to attend to the tones and ignore visual signals, while in the second condition they were asked to attend to the visual signals and ignore the tones. Activity in primary auditory cortex and adjacent areas was strongly correlated with rate of tone presentation, but this relationship was not affected by the direction of attention. In only one area, the right midthalamus, was the response to tones modulated by attention. In this area responses to tones occurred when attention was directed to sound, but not when attention was directed to visual stimuli. There is considerable evidence that the EEG evoked response to tones (N100/Nd response) is strongly modulated by attention and arises in auditory cortex. The ERP is the sum of activity from many sources. The amplitude of this response reflects not only the amount of activity in these sources, but also the degree of synchrony between them. The difference between these typical ERP results and our result from PET could be resolved if we assume that, in our paradigm, attention did not increase the amount of neural activity in auditory cortex, but rather the degree of synchrony between many sources. The signal in the thalamus, which we observed only when volunteers were attending to the tones, might provide the basis for this synchrony. PMID- 9345512 TI - Asymmetry in the human motor cortex and handedness. AB - Handedness is one of the most obvious functional asymmetries, but its relation to an anatomical asymmetry of the hand representation area in the motor cortex has not been demonstrated. This would be a crucial test for the hypothesis of structure-function correlation in cortical motor organization. Using magnetic resonance morphometry, we show for the first time that the depth of the central sulcus is related to handedness. In right-handers, the left central sulcus is deeper than the right, and vice versa in left-handers. Macrostructural asymmetry is complemented by a microstructural left-larger-than-right asymmetry in neuropil volume (i.e., tissue compartment containing dendrites, axons, and synapses) in Brodmann's area 4. These asymmetries suggest that hand preference is associated with increased connectivity (demonstrated by an increased neuropil compartment in left area 4) and an increased intrasulcal surface of the precentral gyrus in the dominant hemisphere. PMID- 9345514 TI - Support and coordination of neuroscience and informatics research in Europe: research in the field of neuroscience under European Union programs. PMID- 9345513 TI - Detecting activations in PET and fMRI: levels of inference and power. AB - This paper is about detecting activations in statistical parametric maps and considers the relative sensitivity of a nested hierarchy of tests that we have framed in terms of the level of inference (voxel level, cluster level, and set level). These tests are based on the probability of obtaining c, or more, clusters with k, or more, voxels, above a threshold u. This probability has a reasonably simple form and is derived using distributional approximations from the theory of Gaussian fields. The most important contribution of this work is the notion of set-level inference. Set-level inference refers to the statistical inference that the number of clusters comprising an observed activation profile is highly unlikely to have occurred by chance. This inference pertains to the set of activations reaching criteria and represents a new way of assigning P values to distributed effects. Cluster-level inferences are a special case of set-level inferences, which obtain when the number of clusters c = 1. Similarly voxel-level inferences are special cases of cluster-level inferences that result when the cluster can be very small (i.e., k = 0). Using a theoretical power analysis of distributed activations, we observed that set-level inferences are generally more powerful than cluster-level inferences and that cluster-level inferences are generally more powerful than voxel-level inferences. The price paid for this increased sensitivity is reduced localizing power: Voxel-level tests permit individual voxels to be identified as significant, whereas cluster-and set-level inferences only allow clusters or sets of clusters to be so identified. For all levels of inference the spatial size of the underlying signal f (relative to resolution) determines the most powerful thresholds to adopt. For set-level inferences if f is large (e.g., fMRI) then the optimum extent threshold should be greater than the expected number of voxels for each cluster. If f is small (e.g., PET) the extent threshold should be small. We envisage that set-level inferences will find a role in making statistical inferences about distributed activations, particularly in fMRI. PMID- 9345515 TI - Neuroinformatics: opportunities across disciplinary and national borders. PMID- 9345516 TI - Molecular mechanisms of synaptic transmission and its regulation: application to models of cognitive functions. PMID- 9345517 TI - The multidimensional database and neuroinformatics requirements for molecular and cellular neuroscience. PMID- 9345518 TI - Recognizing faces by dynamic link matching. PMID- 9345519 TI - From ion channels to networks and behavior: modeling and biological experiments in interaction. PMID- 9345520 TI - Analysis of information processing in the nervous system using a database of identified neurons. PMID- 9345521 TI - Neuroinformatics as explanatory neuroscience. PMID- 9345523 TI - Analyzing 3D images of the brain. PMID- 9345522 TI - What will save neuroscience? PMID- 9345524 TI - Endophrenology: new statistical techniques for studies of brain form. Life on the hyphen in neuro-informatics. AB - The interweaving here of statistics, image analysis, and neuroscience is a fine example of how neuro-informatics is more than the concentration of its constituent disciplines. Each of the component tools of the morphometric synthesis--the deformation model, Procrustes shape coordinates, the thin-plate spline--gains greatly in power in the context of the others, and enhances, too, great recent strides in the instrumentation leading to the raw image data itself (MR scanner physics, multiple stains, new contrast agents). The modern morphometric tool kit seems better matched to the description of gross brain variation than we had any reason to expect. Now the new tools can be exploited to produce remarkably sharper new findings. At the same time, the demands of neuro informatics will press the toolmakers to provide equivalently powerful new techniques in areas presently less developed, such as cortical form or correlations of images with parametric experimental designs or with clinical histories. PMID- 9345525 TI - The developing European computerized human brain database for all imaging modalities. PMID- 9345526 TI - Identifying revolutionary computing technologies. PMID- 9345527 TI - Information management for complex systems: a case study in genomics. AB - Genome mapping and sequencing projects push the frontiers of molecular biology and genetics toward information science. I will try to demonstrate that the information management characteristics of genome research may apply to other large-scale systematic studies of complex systems such as the brain. PMID- 9345529 TI - Linking databases in biotechnology. PMID- 9345528 TI - Electronic collaboratories and digital libraries. PMID- 9345530 TI - Technical foundations and pitfalls of clinical fMRI. AB - Magnetic resonance imaging (MRI) has become an established and invaluable tool in the diagnosis of numerous diseases through its ability to show pathologic contrast in images of soft tissue. More recently, MRI has found application in the study of organ function, principally in the brain and heart. This article deals with MRI imaging of brain function and describes some of the techniques that allow physiological parameters such as cerebral blood volume, cerebral blood oxygenation, and cerebral perfusion to be determined. Additionally, some of the potentially confounding influences in these experiments are discussed. PMID- 9345531 TI - Functional magnetic resonance neuroimaging data acquisition techniques. AB - Functional MRI studies of human brain require rapid data acquisition techniques, to map dynamic changes with sufficient anatomical coverage. In addition, task activation studies employing functional MRI require a high scan sensitivity, in order to discriminate the small, activation-related signal changes from background noise. An overview is given of current fast scan methods, which are sensitized to detect susceptibility changes related to task-induced changes in blood oxygenation. Sources of artifacts are discussed, as well as their effect on image quality. PMID- 9345533 TI - Clinical functional image analysis: artifact detection and reduction. AB - Rapid improvements in functional magnetic resonance neuroimaging technology have resulted in impressive advances in our understanding of structure/function relationships in the human brain. The application of this new technology to the understanding of human brain disease is currently limited by difficulties in extracting task-related signal change from signal intensity time series that have been contaminated by artifacts arising from various intrinsic and extrinsic sources. Effects induced by interscan head motion are a major source of these artifacts. The correction of these artifacts by registration of pairs of reconstructed images has been a focus of research for the past few years and there are now a number of effective means to compensate for this source of noise. This paper discusses issues concerning the prevention and correction of interscan head motion as well as other sources of error variation in fMRI time series. PMID- 9345532 TI - Modeling for intergroup comparisons of imaging data. AB - Intergroup comparisons pose unique challenges in the analysis of functional imaging data. Imperfections in intersubject stereotaxis can give rise to artifactual results and make it particularly important to allow for intersubject differences in task-related changes when formulating statistical models. Because intergroup comparisons generally involve inferences about the populations from which the subjects were drawn rather than inferences about the particular subjects themselves, subjects must be treated as random rather than fixed effects in the statistical model. These requirements, when combined with the need to adjust for multiple spatial comparisons, result in low statistical power when the number of subjects in each group is small. Functional imaging studies to identify differences between groups generally require many more subjects than other types of functional imaging studies and require careful advance planning to maximize the likelihood of reaching meaningful conclusions. PMID- 9345534 TI - Clinical fMRI: implementation and experience. PMID- 9345535 TI - The visual deficit theory of developmental dyslexia. AB - Dyslexia is an impairment in reading that can result from an abnormal developmental process in the case of developmental dyslexia or cerebral insult in the case of acquired dyslexia. It has long been known that the clinical manifestations of developmental dyslexia are varied. In addition to their reading difficulties, individuals with developmental dyslexia exhibit impairments in their ability to process the phonological features of written or spoken language. Recently, it has been demonstrated with a variety of experimental approaches that these individuals are also impaired on a number of visual tasks involving visuomotor, visuospatial, and visual motion processing. The results of these studies, as well as the anatomical and physiological anomalies seen in the brains of individuals with dyslexia, suggest that the pathophysiology of developmental dyslexia is more complex than originally thought, extending beyond the classically defined language areas of the brain. Functional neuroimaging is a useful tool to more precisely delineate the pathophysiology of this reading disorder. PMID- 9345536 TI - fMRI applications in schizophrenia research. AB - fMRI has unique potential in the study of psychiatric patients, particularly in characterizing individual variations and changes over time. We have performed four studies of patients with schizophrenia, using three different fMRI acquisition protocols: (1) 3-D echo-shifted FLASH, a multishot volumetric approach; (2) 3-D PRESTO, a hybid of multishot and echo-planar imaging (EPI) methods that also acquires true volumetric data; and (3) a whole-brain isotropic, multislice EPI technique. Patients were studied during sensorimotor activation and during a novel "N back" working memory paradigm. In general, patients show normal sensorimotor activation responses, although motor cortical activation tends to be less completely lateralized. Prefrontal activation during working memory tends to be reduced in patients with schizophrenia even when performance is normal. A major potential confound in studying this patient population with fMRI is the effect of motion. We propose several methodological standards to address this problem, including comparisons of motion corrections parameters, voxel variances, and the use of an "internal activation standard." PMID- 9345537 TI - Functional MRI and the study of OCD: from symptom provocation to cognitive behavioral probes of cortico-striatal systems and the amygdala. AB - Functional magnetic resonance imaging (fMRI) first appeared in 1991. Since that time there has been a burgeoning use of the technology by psychiatric researchers and neuroscientists. Our group first used fMRI to study obessive compulsive disorder (OCD) with a symptom provocation paradigm and then moved to the use of circuitry-specific cognitive-behavioral probes. The techniques we utilized for the symptom provocation study remain valid today, but have been supplemented by a wide array of new tools. Functional MRI continues to be a rapidly developing technology which could become the gold standard for neuroimaging research in psychiatry. With this in mind, this paper focuses on the past, present, and future applications of fMRI to one model illness, namely OCD. We examine the strengths and limitations of our initial OCD symptom provocation study and then evaluate the use of fMRI with cognitive-behavioral probes of cortico-striatal circuitry and limbic (amygdala) circuitry. We conclude with a brief summary of foreseeable developments which will influence the implementation of fMRI for psychiatric neuroscience in general. PMID- 9345538 TI - Functional MRI applications in clinical epilepsy. AB - Functional MRI holds great promise as a diagnostic tool in presurgical evaluation of patients with epilepsy. Recent research has used fMRI for localization of the seizure focus by tracking interictal spikes and by observing blood flow changes during seizure onset. Localization of the language-dominant hemisphere with fMRI has been well-validated in normal volunteers and left-hemisphere-dominant epilepsy patients, but is less developed for patients with mixed- or right hemisphere language dominance. Intrahemispheric mapping of sensory and motor functions with fMRI is fairly well established, and progress is being made in mapping higher-level cognitive functions, particularly language. Little research has convincingly demonstrated hippocampal function in the normal brain during memory processing, and applications of memory mapping in epilepsy are still lacking. While the clinical use of fMRI in epilepsy needs substantially more study, this is nonetheless a very promising technique that may dramatically change the presurgical diagnostic regimen for these patients. PMID- 9345540 TI - Combining spatial extent and peak intensity to test for activations in functional imaging. AB - Within the framework of statistical mapping, there are up to now only two tests used to assess the regional significance in functional images. One is based on the magnitude of the foci and tends to detect high intensity signals, while the second is based on the spatial extent of regions defined by a simple thresholding of the statistical map, a test that is more sensitive to extended signals. The aim of this paper is to combine the two tests into a single test that is more sensitive to a wider range of signals. This combined test is based on an analytical approximation of the distribution of these two parameters (size and height) and is applied in the context of statistical maps. The risk of error in noise-only 2D or 3D volumes is assessed under a wide range of experimental conditions obtained by varying both the resolution of the map and the threshold at which clusters are defined. In addition, we have investigated this new test on simulated signals, and applied it to an experimental PET dataset. The experimental risk of error is close to the predicted one, and the overall sensitivity increases when analyzing a volume containing different types of signals. PMID- 9345539 TI - Applications of dynamic susceptibility contrast magnetic resonance imaging in Neuropsychiatry. AB - Functional neuroimaging has assumed an important role in the cognitive and clinical neurosciences. Recently, substantial progress has been made toward developing functional magnetic resonance imaging techniques for the examination of cerebral hemodynamic changes that accompany brain function and toward earlier and better diagnosis of brain disease. Dynamic susceptibility contrast (DSC) MRI offers unique information about cerebral hemodynamics both at rest and in response to brain activation. We review the clinical applications of DSC MRI and present our experience with this modality in the evaluation of patients with neuropsychiatric disorders. Our experience suggests that DSC MRI may afford new insights into the diagnosis and treatment of cognitive disorders. PMID- 9345541 TI - Database challenges and solutions in neuroscientific applications. AB - In the scientific community, the quality and progress of various endeavors depend in part on the ability of researchers to share and exchange large quantities of heterogeneous data with one another efficiently. This requires controlled sharing and exchange of information among autonomous, distributed, and heterogeneous databases. In this paper, we focus on a neuroscience application, Neuroanatomical Rat Brain Viewer (NeuART Viewer) to demonstrate alternative database concepts that allow neuroscientists to manage and exchange data. Requirements for the NeuART application, in combination with an underlying network-aware database, are described at a conceptual level. Emphasis is placed on functionality from the user's perspective and on requirements that the database must fulfill. The most important functionality required by neuroscientists is the ability to construct brain models using information from different repositories. To accomplish such a task, users need to browse remote and local sources and summaries of data and capture relevant information to be used in building and extending the brain models. Other functionalities are also required, including posing queries related to brain models, augmenting and customizing brain models, and sharing brain models in a collaborative environment. An extensible object-oriented data model is presented to capture the many data types expected in this application. After presenting conceptual level design issues, we describe several known database solutions that support these requirements and discuss requirements that demand further research. Data integration for heterogeneous databases is discussed in terms of reducing or eliminating semantic heterogeneity when translations are made from one system to another. Performance enhancement mechanisms such as materialized views and spatial indexing for three-dimensional objects are explained and evaluated in the context of browsing, incorporating, and sharing. Policies for providing the system with fault tolerance and avoiding possible intellectual property abuses are presented. Finally, two existing systems are evaluated and compared using the identified requirements. PMID- 9345542 TI - Frequency variation of a pattern-flash visual stimulus during PET differentially activates brain from striate through frontal cortex. AB - We evaluate regional cerebral blood flow (rCBF) in 19 healthy elderly subjects, mean age 64 +/- 11 (SD, years), during a passive visual stimulus in which pattern flash frequency was parametrically manipulated. Using goggles with a grid of red lights imbedded into each lens, we performed five positron emission tomography (PET) H2(15)O water scans on each subject at alternating (left to right eye) flash frequencies of 0, 1, 4, 7, and 14 Hz. We found a biphasic rising and falling rCBF response in the striate cortex (7 Hz peak) and left anterior cingulate (4 Hz peak), 1 Hz activation in left middle temporal gyrus (V5), monotonically increasing rCBF in posterior areas (lateral and inferior visual association areas, Brodmann 18 and 19), and monotonically decreasing rCBF in anterior areas (frontal, cingulate, and superior temporal) predominantly in right hemisphere. We suggest the striate rCBF changes at all frequencies primarily reflect lateral geniculate input, the middle temporal activation at 1 Hz reflects perception of apparent motion, and the posterior extrastriate rCBF monotonic increase represents a neural response to increasing luminance intensity and form and color complexity that occur as pattern-flash frequency increases. The anterior monotonic rCBF decrease may represent active cross-modal functional suppression of brain areas irrelevant for processing the passive visual stimulus. Pattern-flash rCBF responses were highly reproducible (no series effect), more so in posterior than in anterior brain regions. The reproducibility and systematically changing rCBF responses to this passive stimulus suggest that it could be successfully used as a disease probe to evaluate neural function and drug effects in cognitively impaired patients. PMID- 9345543 TI - Visuomotor transformations for reaching to memorized targets: a PET study. AB - Positron emission tomography (PET) was used to identify cortical and subcortical regions involved in the control of reaching to visual targets. Regional cerebral blood flow (rCBF) was measured in eight healthy subjects using H2(15)O PET during the performance of three different tasks. All tasks required central fixation while a 400-ms target was flashed every 5 s at a random location around a virtual circle centered on the fixation target. Additional instructions differed according to the task: (i) visual detection of the target without overt responses; (ii) immediate pointing to the most recent target in the sequence, and (iii) pointing to the previous target in the sequence. By design, the two motor tasks differed in the cognitive processing required. In each trial of immediate pointing, the spatial location of only the most recent target needed to be processed. In each trial of pointing to the previous, instead, while the most recent target was stored in memory for the movement of the next trial, the previous target had to be retrieved from memory to direct the current movement. Limb trajectories were comparable between the two motor tasks in terms of most spatiotemporal parameters examined. Significant rCBF increases were identified using analysis of covariance and t statistics. Compared with visual detection there was activation of primary sensorimotor cortex, ventrolateral precentral gyrus, inferior frontal gyrus in the opercular region, supramarginal gyrus, and middle occipital gyrus, all these sites in the hemisphere (left) contralateral to the moving limb, and cerebellar vermis, during both immediate pointing and pointing to the previous. During immediate pointing there was additional activation of left inferior parietal lobule close to the intraparietal sulcus, and when compared with pointing to the previous, dorsolateral prefrontal cortex bilaterally. During pointing to the previous, instead, there was additional activation of supplementary motor cortex, anterior and midcingulate, and inferior occipital gyrus in the left hemisphere; superior parietal lobule, supramarginal gyrus, and posterior hippocampus in the right hemisphere; lingual gyri and cerebellar hemispheres bilaterally; anterior thalamus; and pulvinar. The activation of two partially distinct cerebral networks in these two motor tasks reflects the different nature of signal processing involved. In particular, the specific activation of intraparietal sulcus and prefrontal cortex in immediate pointing appears characteristic of a network for visuospatial working memory. By contrast, the corticolimbic network engaged in pointing to the previous could mediate spatial attention and the sequence of encoding, recording, and decoding of spatial memories required by a dual task with two competing targets. PMID- 9345544 TI - Mapping histology to metabolism: coregistration of stained whole-brain sections to premortem PET in Alzheimer's disease. AB - The association between [18F]fluorodeoxyglucose positron emission tomography (FDG PET) counts obtained 8 h before death and neurofibrillary tangle (NFT) staining density in a patient with Alzheimer's disease (AD) was evaluated. In our patient FDG-PET counts were globally decreased with a greater focal deficit in the left medial temporal region independent of volume loss. After death, whole-brain sections derived from cryomacrotome sectioning were stained for NFTs by the Gallyas method and elastically warped into their native space enabling registration with premortem FDG-PET data. Gallyas staining density was localized to the paralimbic cortex of the basal forebrain, medial temporal, and orbital frontal regions. The poor correlation between NFT staining density and hypometabolism on FDG-PET implicates alternate mechanisms underlying the metabolic defect in AD. PMID- 9345545 TI - Comparison of fluorescent voltage-sensitive dyes for multisite optical recording in hamster cerebral cortex by measurement of bicuculline-induced epileptiform events. AB - Two fluorescent voltage-sensitive dyes, RH795 and DI-2-ANEPPQ, were compared for in vivo multisite optical recording from the gustatory insular cortex of the golden Syrian hamster, the first reported use of DI-2-ANEPPQ in a mammalian brain preparation. The exposed cortex of the anesthetized hamster was stained with a 500 microM solution of either dye, and the cortical surface was imaged onto a 124 element photodiode array by an epifluorescence microscope equipped with a 2.5x objective (0.08 na) and appropriate excitation and emission filters for each dye. Background fluorescence was recorded with amplifiers in DC-coupled mode, and the bathing solution changed to one containing 100 microM bicuculline methiodide. Large, widespread epileptiform events were recorded optically, and with a surface electrode, within 2 min. Three experiments were carried out with each dye. The 10 recorded events from each experiment (and five detector records from each of these 10) having the largest signals were selected for comparison. Five measures were derived from the data: (1) The ratio of peak signal fluorescence to background fluorescence (delta F/F), (2) signal-to-noise power ratio (S/N), (3) root mean square noise (RMS noise), (4) an amplitude ratio (AR) consisting of the signal height divided by RMS noise, and (5) the peak value of the surface electrode record (SER). The results indicate a twofold increase in delta F/F and a fivefold increase in S/N (twofold increase in AR) with DI-2-ANEPPQ. No significant difference was found between dyes in the RMS noise or SERs. In addition, signals did not decline detectably over time with DI-2-ANEPPQ, but declined about 25%/h with RH795. PMID- 9345546 TI - Transients, metastability, and neuronal dynamics. AB - This paper is about neuronal dynamics and how their special complexity can be understood in terms of nonlinear dynamics. There are many aspects of neuronal interactions and connectivity that engender the complexity of brain dynamics. In this paper we consider (i) the nature of this complexity and (ii) how it depends on connections between neuronal systems (e.g., neuronal populations or cortical areas). The main conclusion is that simulated neural systems show complex behaviors, reminiscent of neuronal dynamics, when these extrinsic connections are sparse. The patterns of activity that obtain, under these conditions, show a rich form of intermittency with the recurrent and self-limiting expression of stereotyped transient-like dynamics. Despite the fact that these dynamics conform to a single (complex) attractor this metastability gives the illusion of a dynamically changing attractor manifold (i.e., a changing surface upon which the dynamics unfold). This metastability is characterized using a measure that is based on the entropy of the time series' spectral density. PMID- 9345547 TI - Anterior cingulate and motor network metabolic impairment in progressive supranuclear palsy. AB - Progressive supranuclear palsy is the prototype of subcortical dementia. Using positron emission tomography and statistical parametric mapping, we compared the glucose metabolic pattern obtained in this subcortical dementia to that observed in elderly healthy controls and in Alzheimer's disease, the prototype of cortical dementia. Progressive supranuclear palsy was characterized by a relative decrease of metabolism in anterior cingulate, adjacent supplementary motor area, precentral cortex, middle prefrontal cortex, midbrain tegmentum, globus pallidus, and ventrolateral and dorsomedial nuclei of thalamus. The data in progressive supranuclear palsy highlight predominant metabolic impairment in brain structures engaged in response selection, in attention for action, and in motor networks. PMID- 9345548 TI - Empirical analyses of BOLD fMRI statistics. I. Spatially unsmoothed data collected under null-hypothesis conditions. AB - Temporal autocorrelation, spatial coherency, and their effects on voxel-wise parametric statistics were examined in BOLD fMRI null-hypothesis, or "noise," datasets. Seventeen normal, young subjects were scanned using BOLD fMRI while not performing any time-locked experimental behavior. Temporal autocorrelation in these datasets was described well by a 1/frequency relationship. Voxel-wise statistical analysis of these noise datasets which assumed independence (i.e., ignored temporal autocorrelation) rejected the null hypothesis at a higher rate than specified by the nominal alpha. Temporal smoothing in conjunction with the use of a modified general linear model (Worsley and Friston, 1995, NeuroImage 2: 173-182) brought the false-positive rate closer to the nominal alpha. It was also found that the noise fMRI datasets contain spatially coherent time signals. This observed spatial coherence could not be fully explained by a continuously differentiable spatial autocovariance function and was much greater for lower temporal frequencies. Its presence made voxel-wise test statistics in a given noise dataset dependent, and thus shifted their distributions to the right or left of 0. Inclusion of a "global signal" covariate in the general linear model reduced this dependence and consequently stabilized (i.e., reduced the variance of) dataset false-positive rates. PMID- 9345549 TI - Empirical analyses of BOLD fMRI statistics. II. Spatially smoothed data collected under null-hypothesis and experimental conditions. AB - In the companion to this paper (E. Zarahn, G. K. Aguirre, and M. D'Esposito, 1997, NeuroImage, 179-197), we describe an implementation of a general linear model for autocorrelated observations in which the voxel-wise false-positive rates in fMRI "noise" datasets were stabilized and brought close to theoretical values. Here, implementations of the model are tested for use with statistical parametric mapping analysis of spatially smoothed fMRI data. Analyses using varying models of intrinsic temporal autocorrelation and either including or excluding a global signal covariate were conducted upon human subject data collected under null hypothesis as well as under experimental conditions. We found that smoothing with an empirically derived impulse response function (IRF), combined with a model of the intrinsic temporal autocorrelation in spatially smoothed fMRI data, resulted in a map-wise false-positive rate which did not exceed a 5% level when a nominal alpha = 0.05 tabular threshold was applied. Use of other models of intrinsic temporal autocorrelation resulted in map-wise false positive rates that significantly exceeded this level. fMRI data collected while subjects performed a behavioral task were used to examine (a) task-dependent global signal changes and (b) the dependence of sensitivity on the temporal smoothing kernel and inclusion/exclusion of a global signal covariate. The global signal changes within an fMRI dataset were shown to be influenced by the performance of a behavioral task. However, the inclusion of this measure as a covariate did not have an adverse affect upon our measure of sensitivity. Finally, use of an empirically derived estimate of the IRF of the system was shown to result in greater map-wise sensitivity for signal changes than the use of a broader (in time) Poisson (parameter = 8 s) kernel. PMID- 9345550 TI - Another neural code? AB - This paper presents the conjecture that functional integration may be mediated by the mutual induction and maintenance of stereotyped spatiotemporal patterns of activity (i.e., transients) in different neuronal populations. In contradistinction to temporal and rate coding models of neuronal interactions, transient coding considers that transactions among neuronal systems use transient dynamics that are distributed in a structured way over both space and time. In contrast to synchronization models, transient coding does not depend on interactions at the same frequencies, in different parts of the brain, but involves covariations among different frequencies and can therefore be considered a more general form of coding. Using an analysis of the correlations among the spectral density of neuromagnetic signals, measured at different cortical regions, this hypothesis was confirmed. For example high (gamma)-frequency oscillations in the prefrontal cortex are associated with low (20 Hz)-frequency oscillations in the parietal cortex. The results are consistent with transient coding and suggest that transient dynamics endure for at least 40-200 ms. Transient coding means that correlations (rate coding) and coherence (synchrony) are neither complete nor sufficient characterizations of neuronal interactions. Although temporal coding, rate coding, and synchrony are important aspects of neuronal interactions, the results speak to further integrative neuronal mechanisms of a more general nature. PMID- 9345551 TI - Modulation of human cortical rolandic rhythms during natural sensorimotor tasks. AB - We studied modulation of cortical neuromagnetic rhythms in association with left and right median nerve stimulation, during rest, finger movements, and passive tactile hand stimulation, in seven healthy, right-handed adults. In the rest condition, the amplitude of the rhythmic sensorimotor activity decreased immediately after the median nerve stimuli and increased above the prestimulus level within 0.4 s afterward, especially in the 7- to 25-Hz band. The rebound occurred 100-300 ms earlier for 20 (7-15)-than for 10 (15-25)-Hz activity. Suppressions and rebounds were strongest in the contralateral sensorimotor hand area for the 20-Hz, but not for the 10-Hz, activity. The maximum rebound was on average 22-34% stronger in the left than in the right hemisphere. Active exploration of objects abolished rebounds of both 10- and 20-Hz signals in the contralateral hemisphere and markedly diminished them ipsilaterally. Finger movements without touching an object and passive tactile stimulation produced a weaker effect. The sensorimotor rhythms thus show a characteristic suppression and subsequent rebound after electrical median nerve stimulation. The rebound is left-hemisphere dominant in right-handed subjects and its suppression reveals bilateral cortical activation during both motor tasks and passive tactile stimulation, especially for explorative finger movements. PMID- 9345552 TI - Cognitive subtractions may not add up: the interaction between semantic processing and response mode. AB - Determining the areas of brain activity associated with cognitive processing has typically relied on the use of a subtraction paradigm, which is based on the premise that the neural processes underlying behavior are additive. If the additivity assumption is valid then brain regions associated with a semantic processing task should be the same regardless of how participants make a response. To investigate this proposition, participants underwent six PET scans, in which they made semantic or letter word judgments, responding "yes" or "no" in three different modes: mouse-clicking, spoken response, or silent thought. Analyses showed an increase in regional cerebral blood flow associated with semantic processing in the left inferior frontal cortex, anterior cingulate, and right cerebellum for all three response conditions. However, there was a significant interaction: the greatest increase was observed in the mouse-click condition and the weakest change seen with silent thought. Moreover, other areas of the brain were uniquely activated for each response mode. The results indicate that different areas of the brain were recruited for semantic processing depending on how participants had to organize their responses. Implications for the additivity assumption and methods of analysis to be used in conjunction with the subtraction technique are discussed. PMID- 9345553 TI - Activation of human V5 complex and rolandic regions in association with moving visual stimuli. AB - We recorded magnetoencephalographic responses from seven healthy humans during the presentation of stationary and rotating radial gratings. Rotations lasting 1 s evoked movement-specific sustained activity in the parieto-occipitotemporal border area, in agreement with the activation of the V5 complex specialized for the analysis of movement. The source areas of the movement-specific sustained fields were transiently active 100-130 ms after the onsets of both rotating and stationary stimuli, suggesting that movement-related cortical areas respond to any transient changes in the visual environment. Transients were evoked also in other brain areas 60-200 ms after onsets of both stimuli. Four subjects displayed additional motion-related sustained activity in the rolandic region. Sustained activity continued after the stimulus movement in several subjects during perception of the movement aftereffect. The transient activity may evoke visual attention while sustained activity of the V5 complex may be related to the conscious perception of movement. PMID- 9345554 TI - In vivo evidence for the involvement of dopamine-D2 receptors in striatum and anterior cingulate gyrus in major depression. AB - The dopaminergic system is a candidate neurotransmitter system thought to be involved in the pathogenesis of depression. This study addresses the issue whether the antidepressant efficacy of serotonin reuptake inhibition is related to changes in the cerebral dopaminergic system. Cerebral dopamine-D2 receptors were characterized in 13 patients with major depression using the dopamine-D2 receptor antagonist iodobenzamide and single photon emission tomography. Dopamine receptor binding was assessed twice, before and during serotonin reuptake inhibition. An increase in dopamine-D2 receptor binding during serotonin reuptake inhibition was found in striatum and anterior cingulate gyrus in treatment responders, but not in nonresponders. The increase in dopamine-D2 receptor binding correlated significantly with clinical recovery from depression as assessed with the Hamilton depression scale (r = 0.59 for right and left striatum respectively, P < 0.05; r = 0.79 for the anterior cingulate gyrus, P < 0.05 after Bonferroni correction). Qualitatively similar correlations were observed in the precentral gyrus, the medial frontal gyrus, the inferior frontal gyrus, and the frontal part of the opercular gyrus, but these correlations failed to reach statistical significance after correction for the effects of multiple testing. No such correlations were found in the superior frontal gyrus, the orbitofrontal gyrus, the gyrus rectus, the superior parietal gyrus, or the superior temporal gyrus. The data strengthen the concept that the striatum and the anterior cingulate gyrus are involved in mood regulation. Dopamine-D2 receptors may constitute a central role in this domain. PMID- 9345555 TI - Cognitive conjunction: a new approach to brain activation experiments. AB - This paper introduces the concepts and procedures of "cognitive conjunction," a new approach to designing and analyzing cognitive activation experiments. Cognitive conjunction compliments categorical approaches such as cognitive subtraction and requires a specific form of statistical inference that involves the conjunction of several hypotheses. While cognitive subtraction studies are designed such that a pair of tasks differ only by the processing component(s) of interest, cognitive conjunction studies are designed such that two or more distinct task pairs each share a common processing difference. The neural correlates of the process of interest are then associated with the common areas of activation for each task pair. There are two main advantages of cognitive conjunction relative to cognitive subtraction. The first is that it provides a greater latitude for selecting baseline tasks because it is not necessary to control for all but the component of interest. The only constraint on selecting the baseline is that the component of interest is the only process that differs in each task pair. The second advantage is that cognitive conjunction does not depend on "pure insertion"--the assumption that the addition of an extra processing component in the activation task has no effect on the implementation of processes that are also engaged by the baseline task. The differences between the design and the statistical analysis of experiments based on cognitive subtraction, cognitive conjunction, and factorial designs are illustrated with a study of phonological retrieval. Cognitive conjunction analysis indicates that irrespective of whether subjects name words, objects, letters, or colors, there is activation of the left posterior basal temporal lobe, the left frontal operculum, the left thalamus, and the midline cerebellum. PMID- 9345556 TI - MRI and PET coregistration--a cross validation of statistical parametric mapping and automated image registration. AB - Coregistration of functional PET and T1-weighted MR images is a necessary step for combining functional information from PET images with anatomical information in MR images. Several coregistration algorithms have been published and are used in functional brain imaging studies. In this paper, we present a comparison and cross validation of the two most widely used coregistration routines (Friston et al., 1995, Hum. Brain Map. 2: 165-189; Woods et al., 1993, J. Comput. Assisted Tomogr: 17: 536-546). Several transformations were applied to high-resolution anatomical MR images to generate simulated PET images so that the exact (rigid body) transformations between each MR image and its associated simulated PET images were known. The estimation error of a coregistration in relation to the known transformation allows a comparison of the performance of different coregistration routines. Under the assumption that the simulated PET images embody the salient features of real PET images with respect to coregistration, this study shows that the routines examined reliably solve the MRI to PET coregistration problem. PMID- 9345557 TI - Factors that influence effect size in 15O PET studies: a meta-analytic review. AB - The PET literature is growing exponentially, creating a need and an opportunity to perform a meta-analytic review consolidating the published information. This study describes the use of effect size as an index in PET studies and discusses how this measure can be used for comparing findings across studies, laboratories, and paradigms. In comparing studies across laboratories it is essential to know how the methods employed affect the results and conclusions drawn. This study also compared effect size for two different methods of tracer delivery in 15O PET studies ([15O]H2O bolus injection versus inhalation of [15O]CO2), whether averaged versus single-scan conditions were used, and the data analytic strategy employed. The effect sizes observed across studies were consistently large with a median effect size of 8.55, indicating that the phenomena investigated in 15O PET studies are strong. The largest peak activation reported in a study was found to be affected by variability in sample size, data analytic strategy, and repeat versus single-scan conditions. However, the impact of these factors was not examined on smaller or less intense peaks. Minimal standards for reporting statistical results are discussed. PMID- 9345558 TI - Near-infrared optical detection of sequential brain activation in the prefrontal cortex during mental tasks. AB - To examine the spatiotemporal differences of brain activation during mental tasks, changes in the oxygenation and hemodynamics in two regions of the prefrontal cortex were measured simultaneously by near-infrared spectroscopy (NIRS). Subjects were eight healthy adults who attempted to solve three different mathematical problems. The behavior of concentration changes in oxy-, deoxy-, and total hemoglobin in one brain region varied with the time course (more than 10 min). This suggested that regional brain activity varied during the performance of the mental task. In each single subject, the pattern of these changes varied with each problem, and this variation differed from subject to subject. When NIRS traces in two regions were compared, it was seen that activated regions moved alternatively: when in one region total hemoglobin that had first increased returned to the resting level, in the other it started to increase. These region dependent temporal variations of brain activity might reflect mental processes. It is thus concluded that NIRS has the potential for imaging the sequence of brain activation. PMID- 9345560 TI - [Chromosome mosaicism and intrauterine growth retardation]. PMID- 9345559 TI - Simultaneous recording of epileptiform discharges by MEG and subdural electrodes in temporal lobe epilepsy. AB - Spontaneous epileptiform discharges were recorded by whole head magnetoencephalography (MEG) and subdural electrodes simultaneously from two patients with medically intractable temporal lobe epilepsy. In one patient whose epileptiform discharges emerged from the lateral temporal lobe, simultaneously recorded MEG could estimate equivalent current dipole reliably near the tumor. The amplitude of the dipole was in proportion not only to the amplitude of epileptiform discharge but also to the number of subdural electrodes involved. In the other patient, MEG detected only a small proportion of epileptiform discharges, even when they were recorded by subdural electrodes from the mesial temporal lobe. It is concluded that the amplitude and the depth of epileptiform discharges would largely affect the sensitivity of dipole localization by MEG. PMID- 9345561 TI - [Study of plasma acylcarnitines using tandem mass spectrometry. Application to the diagnosis of metabolism hereditary diseases]. AB - BACKGROUND: L-carnitine is known to transport long chain fatty acids through the mitochondrial membrane but also to export accumulated acyl-CoA's as acylcarnitine esters. Acylcarnitine identification in body fluids allows the diagnosis of mitochondrial inborn errors especially fatty oxidation defects. Tandem mass spectrometry represents a new method for isolation and identification of acylcarnitines in plasma or in blood spotted onto filter paper (Guthrie cards). MATERIAL AND METHODS: In order to validate our method, we studied 30 plasmas from children affected with 15 different inborn errors of metabolism and five amniotic fluids from fetuses affected with several organic acidurias. Fourty-six samples from children at risk for mitochondrial fatty oxidation disorders have been analyzed. We developed a method of tandem mass spectrometry with liquid secondary ion mass spectrometry using deuterated acylcarnitines as internal standards. RESULTS: This method is very sensitive (detection limit = 2 microM). In all affected patients specific acylcarnitine signals corresponding to the metabolic block were constantly found. This confirms the diagnosis and validates the method. Among the 46 at risk children, four defects of long chain fatty acid oxidation were identified. CONCLUSION: This new method is of great interest especially for the long chain fatty acid oxidation defects. These defects are very difficult to diagnose with classical methods as urinary organic acid profiling. A small amount of plasma (100 microL) or blood spotted onto paper is required. The acylcarnitine profile allows a rapid diagnosis if a dedicated apparatus is available. PMID- 9345562 TI - [Primary immunodeficiency in Tunisia: study of 152 cases]. AB - BACKGROUND: Primary immunodeficiencies are rare immunopathological disorders. A multidisciplinary study group was set up in Tunis in 1988 and has since identified 152 cases of such diseases. We herein present our series and compare it to the international registries. POPULATION AND METHODS: Over a period of 8 years (April 1988-April 1996), 295 children suffering from recurrent infections were investigated; primary immunodeficiency was confirmed in 152 out of them. The immunological investigation included a study of specific and/or non specific humoral and cellular immunity. RESULTS: These 152 patients belonged to 129 families among which 70 were consanguine (54%). Familial primary immunodeficiency occurred in 23 of them. In 39 families (30%), one or more deaths occurred during early childhood. In more than half of the cases (89 cases), the immunological investigations revealed a cellular or combined immunodeficiency with a majority of ataxia-telangiectasia syndromes (53 cases), T cell activation immunodeficiencies (12 cases) and HLA class II deficiency (nine cases). A predominant antibody defect was observed in 35 patients with a majority of agammaglobulinemia (11 cases) and hyper-IgM syndromes (11 cases). A defect of non specific cellular immunity was found in 18 cases (11.8%) including seven cases of chronic granulomatous disease and five cases of leukocyte adhesion deficiency. Three children (1.9%) were deficient in the complement system. Deaths occurred so far in 37 patients (24.3%). CONCLUSIONS: Primary immunodeficiencies are relatively frequent in Tunisia, probably because of the high rate of consanguinity among the general population. The distribution of the different groups of primary immunodeficiencies is characterized by high frequency of ataxia telangiectasia and hyper-IgM syndrome and scarcity of severe combined immunodeficiencies and Wiskott-Aldrich syndrome. PMID- 9345563 TI - [Evaluation of the ambulatory treatment of acute diarrhea in infants. Reseau interhospitalier d'evaluation des pratigues medicales dans les affections courantes de l'enfant]. AB - BACKGROUND: Gastroenteritis remains a common and expensive illness. Oral rehydration solutions (ORS) have been shown to be effective in the prevention and treatment of dehydration, the prime cause for diarrhea-related morbidity and mortality. OBJECTIVE: To evaluate the ambulatory management of acute gastroenteritis in infants, and particularly practices concerning oral fluid therapy. METHODS: This prospective, multicenter study included 326 infants (mean age: 10 +/- 6 months), examined in a hospital for acute gastroenteritis, with or without dehydration. RESULTS: Before admission, 81% had previously been examined by a practitioner, and 89% of these practitioners had written a prescription. This prescription included ORS in 35% and was not different according to the age of the infant. Pediatricians prescribed ORS more frequently than general practitioners (respectively 58% vs 29%; P < 0.001). The failure rate of ORS prescription was 25% (two parents did not observe, ten children refused to drink, and eight stopped treatment because of vomiting). Lactose-free milks were prescribed in 46% of infants and the observance was 82%. At least one drug was prescribed in 94% of infants, with a mean of 2.6 drugs per infant; one antibiotic was prescribed in 33% of infants. Infants were admitted to hospital without any previous consultation in 18%, on the parents' initiative but after at least one previous medical examination in 52%, and on the physician's initiative in 30%. Thirty-three percent were dehydrated; one infant died and two had sequellae. CONCLUSION: The use of ORS remains insufficient. Efforts to improve use of ORS should be expanded beyond physician education. PMID- 9345564 TI - [Sickle cell anemia in children: value of hydroxyurea in severe forms]. AB - BACKGROUND: Sickling of red cells in patients with sickle cell anemia causes painful vaso-occlusive crises (VOC). Hydroxyurea (HU) has been shown to increase HbF production and therefore has the potential to prevent these crises in adult patients. This work aimed to confirm its clinical efficiency in children. PATIENTS AND METHODS: Since 1993, eight children and adolescents (5-16 years old) with hemoglobinopathy (HbS > 65%) were given HU (14 to 27 mg/kg/day) for a mean duration of 10 months. We did a retrospective study about different data, especially inpatient days for VOC, pain intensity during these crises, individual transfusion requirements and HbF level. RESULTS: As the HbF levels increased, we observed a reduction of the inpatient days for VOC, of the pain intensity during these crises and of the mean number of units transfused per month. CONCLUSION: HU is the first clinically acceptable drug shown to prevent painful crises in adults but also in children with sickle cell anemia. However, its effects and potential toxicity are still unknown and the other therapeutic possibilities have to be considered before starting a long-term treatment with HU. PMID- 9345565 TI - [Acute respiratory insufficiency revealing myasthenia gravis: apropos of 3 cases]. AB - BACKGROUND: Myasthenia gravis is usually revealed by a ptosis or a diplopia. A respiratory muscle weakness often occurs during the course but an acute respiratory failure as initial feature is unusual. CASE REPORTS: Three girls, aged 8, 10 and 14 years, were hospitalised in an intensive care unit, along a 15 year-period, for an acute respiratory distress. The first two children suffered from skeletal and bulbar muscle weakness. The third, admitted with the diagnosis of unexplained pneumonia, was complaining of skeletal and bulbar muscle weakness for the last 18 months. Myasthenia gravis was confirmed with electromyography, and detection of the acetylcholine-receptors antibodies in all three cases. CONCLUSION: Any unexplained acute respiratory distress must lead to search for skeletal and bulbar muscle weakness, specially after muscular exercise or at the end of day, manifestations which characterize myasthenia gravis. PMID- 9345566 TI - [Superior sagittal sinus thrombosis and nephrotic syndrome: favorable outcome with low molecular weight heparin]. AB - BACKGROUND: Nephrotic syndrome is known to be associated with thrombosis but rarely of cerebral vessels. CASE REPORT: A 3-year old child with steroid dependent nephrotic syndrome was hospitalized for drowziness followed by a left hemiparesis. The CTscan showed a superior sagittal sinus thrombosis. The child completely recovered after treatment by low molecular weight heparin (LMWH). CONCLUSION: LMWH could be used for preventing and/or treating thrombosis associated with nephrotic syndrome. Nevertheless, controlled studies are necessary for assessing its efficacy and absence of risk in children. PMID- 9345567 TI - [Acute pyelonephritis and subcapsular hematoma revealing nephroblastoma at the age of 15 years]. AB - BACKGROUND: Nephroblastoma' the most common renal tumor in children between 1 and 5 years, occurs rarely in the oldest child. CASE REPORT: A 16-year-old teenager suffered from acute pyelonephritis caused by Klebsiella pneumoniae. Renal ultrasonography showed a left subcapsular hematoma; the CT scan confirmed the finding and also showed renal scarring. However, a second CT scan showed pulmonary nodules suggestive of metastasis, a diagnosis that was confirmed by needle biopsy of pulmonary lesions. Recovery was obtained after chemotherapy and nephrectomy with a 3-year-follow-up. CONCLUSION: This nephroblastoma was particular because its development in an adolescent, its association with acute pyelonephritis and subcapsular hemorrhage. PMID- 9345568 TI - [Urogenital tuberculosis in children]. AB - BACKGROUND: Tuberculosis of the uro-genital tract is uncommon in both sexes before puberty. CASE REPORTS: Case 1. A 10 year-old boy suffered from chronic cystitis. Urine examination showed pyuria without bacteria and absence of mycobacterium tuberculosis on direct smears and culture. Intravenous pyelography showed left ureterohydronephrosis and a non functioning right kidney with calcifications. A right nephrectomy was performed which showed caseous lesions with granulomas. A triple specific therapy failed to completely cure the vesicoureteral lesions. Case 2. A 9 year-old boy was admitted for the suspicion of appendicitis. Renal ultrasonography showed features of abscesses located to the right kidney and the liver. Laparotomy showed their tuberculous origin which was confirmed by histological examination and positive search for Mycobacterium tuberculosis in urine. The course was favorable under specific treatment. CONCLUSIONS: Diagnosis of tuberculosis of the uro-genital tract may be difficult leading to a delayed treatment and sequellae. PMID- 9345569 TI - [Imerslund's disease. Clinical and biological aspects. Apropos of 6 cases]. AB - BACKGROUND: Imerslund syndrome, a recessive autosomal disease, initially described by Imerslund and Grasbeck in 1960, associates megaloblastic anemia and proteinuria. CASE REPORT: We report on six cases, studied in five different families. All patients (mean age: 3.5 years) had clinical symptoms of anemia, three had malabsorption, proteinuria was present in five, at the time of diagnosis. Hemogram and decreased serum vitamin B12 levels were consistent with the diagnosis in all cases. Intra-muscular injections of cyanocobalamine was instituted on a life-time basis and the long term prognosis is good. CONCLUSION: The diagnosis should be evoked when the three typical features are present: macrocytic anemia, decreased serum B12 level and proteinuria. It will be confirmed by the bone marrow megaloblastic aspects and the Schilling test findings. PMID- 9345570 TI - [Teratogenic effects of vitamin A and its derivates]. AB - Retinoids, the synthetic derivates of vitamin A, have a key role on cellular differentiation and developmental tissue specificity. Their effects are mediated by nuclear receptors which transactivate homeobox genes. They are teratogenic to animals and they all induce similar malformations dependent on the dose and the duration of exposure. This is a review of the teratogenic effects of vitamin A and its synthetic derivates--isotretinoin, acitretine and topical retinoids--in humans. High dose vitamin A have a potent teratogenic effect and are therefore contra-indicated during pregnancy. Isotretinoin is responsible for a syndrome including malformations of the central nervous system, heart and thymus, together with craniofacial defects. The incidence rate is high and comparable to thalidomide (ie, 25%). This high teratogenic potency justifies a strict limitation of such a prescription in women susceptible to become pregnant. Acitretine, which replaces etretinate because of its long half life of 120 days, might also be teratogenic in humans. In addition, it may be back transformed into etretinate, thus contraindicating pregnancy for 2 years after withdrawal. Finally, despite a low percutaneous resorption, available data on the use of retinoids as topicals are limited and their use during pregnancy is therefore not recommended. Although they are efficient in skin diseases, the use of retinoids in women of the child bearing age is very limited because of their potent teratogenic effect. PMID- 9345571 TI - [Treatment of neonatal convulsions]. AB - Three important measures are concommitantly recommended in the treatment of neonatal seizures. 1) The maintenance of vital functions by supplying oxygen and using mask ventilation in addition to cardiac monitoring and placing an intravenous line. 2) The search for an etiology through metabolic and infectious laboratory tests. In many cases this will lead to an etiological treatment. 3) The specific treatment of the seizures: intravenous or intrarectal diazepam is the first line treatment. Phenobarbital, then phenytoin and clonazepam are used when diazepam is unsuccessful. Indications and duration of a secondary prophylactic treatment are controversial and must be discussed on the basis of the electroencephalographic data and the risk factors for epileptic seizures during the neonatal period and at 3 months of age. PMID- 9345572 TI - [Idiopathic disseminated infection by BCG or atypical mycobacteria]. AB - Disseminated Bacille Calmette-Guerin (BCG) infection following vaccination with live BCG may occur in children without any well-defined immune deficiency. Children with unexplained disseminated infection due to environmental non tuberculous mycobacteria (NTM) have also been reported. Several lines of evidence suggested that such idiopathic BCG and NTM infections may represent different complications of the same autosomal recessive inherited immune disorder. Accordingly, gamma-interferon receptor deficiency was recently identified in three kindreds with NTM and BCG disseminated infection. This article reports the current knowledge on idiopathic BCG or NTM disseminated infections, and the underlying inherited immune deficiencies. PMID- 9345573 TI - [Psychological effects of parental separation on children]. AB - A study on the psychological consequences of parental separation on children was performed among 3,098 pupils of first year secondary school during the 1995-1996 academic year in the department of Isere. The children family situation was compared with the results of personality test of Coopersmith (SEI). The main characteristics of the children, in particular family condition such as parental death or parental separation are in agreement with the French national data. Among the children with separated parents, 29% were less than 3 years old when their parents broke up. In 85% of the cases the child lived mainly with his/her mother, in 9% of the cases with his/her father, in 4% of cases in a joint-custody arrangement, in 1% of the cases in an other person's home. The overall SEI scores were good. There was a significant overrepresentation of girls with extreme scores. Compared with the scores of the children of unseparated parents, the average SEI scores were low for children with separated parents and for children with one dead parent, but differences were observed between girls end boys. The age of children at the time of the separation did not influence the SEI score and there was little influence of the father-child regular contacts. PMID- 9345574 TI - [Radiological case of the month. Gorham disease of costovertebral localization]. PMID- 9345576 TI - [Meckel's diverticulum and acute intestinal intussusception in older children]. PMID- 9345575 TI - [Deep capillary hemangioma of orbit in an infant]. PMID- 9345577 TI - [Moebius syndrome in newborn infants]. PMID- 9345578 TI - [Burns by hot tap water: news and methods of prevention]. PMID- 9345579 TI - [Partial 3q-trisomy (q22-qter) monosomy 13q (q32-qter) by translocation between paternal chromosome 3 and 13]. PMID- 9345580 TI - [Orbital regional anesthesia for postoperative analgesia after eye enucleation in children]. PMID- 9345581 TI - [Anti-phospholipids and vena cava thrombosis in pediatrics]. PMID- 9345582 TI - [Weight and height of 6-year-old schoolchildren in Haute-Garonne in 1991 and Sempe's references]. PMID- 9345583 TI - [Congenital candidiasis: early diagnosis and treatment with fluconazole]. PMID- 9345584 TI - [Is surgery of popliteal cysts in children necessary? Study of a series of 128 cases]. PMID- 9345585 TI - Positron emission tomography: background, possibilities and perspectives in neuroscience. AB - Positron Emission Tomography (PET) is a method for determining biochemical and physiological processes in vivo in a quantitative way. This includes the measurement of the pharmacokinetics of labeled drugs and the measurement of the effects of drugs and/or therapy on metabolism. Also deviations of normal metabolism can be measured and insight in biological processes responsible for diseases can be obtained. To accomplish these measurements a multi-disciplinary team is necessary for the expertise in the fields of radionuclide production, radiochemistry, pharmacy, medicine, nuclear imaging and data analysis. PMID- 9345586 TI - Recent and future evolutions in NeuroSPECT with particular emphasis on the synergistic use and fusion of imaging modalities. AB - Recent and future evolutions in neuroSPECT apply to radiopharmaceuticals techniques and the synergistic use of different imaging modalities in the work-up of neurological disorders. The introduction of Technetium labelled perfusion tracers, which could pass the intact blood-brain barrier, together with the implementation of the tomographic principle, by making the conventional gamma camera rotating, enabled estimation of regional cerebral blood flow and indirectly of local brain metabolism. In addition at present Thallium-201 and Tc 99m sestaMIBI allow functional detection of viable tumor tissue, without interference from previous surgery or radiotherapy as seen using CT-scan or MRI. In neurology this has led to the recognition of SPECT by the American Academy of Neurology (Therapeutics and technology subcommittee) as an established or promising tool in major neurological disorders such as dementia, stroke and epilepsy, while other domains such as brain oncology are considered investigational. With regard to radiopharmaceuticals, recent evolutions mainly include the development of mostly Iodine-123 labelled receptor ligands, some of which are already commercially available. For instrumentation advances consist e.g. of multidetector systems equipped with fanbeam collimators, attenuation and scatter correction or coincidence detection. Given the present role for nuclear neurology it may be expected that these additional radiopharmaceutical and technical innovations will continue to stimulate the development of SPECT of the brain. The synergistic use of several imaging techniques such as CT, (functional) MRI, source imaging, SPECT and PET represents a multimodal holistic approach to probe cerebral functions for research and clinical purposes. Clinical indications, in which this synergistic use is illustrated include e.g. support of the clinical diagnosis of dementia of the Alzheimer type, presurgical ictal detection of seizure focus, detection of acute ischemia and differential diagnosis between radiation necrosis and brain tumor recurrence. The synergistic use of imaging modalities, optimally applied using image fusion, allows to overcome the intrinsic limitations and to enhance the specific advantages of the different approaches as it leads to increased precision and accuracy, as well for spatial anatomofunctional correlation as for quantification. PMID- 9345587 TI - Is positron emission tomography useful in stroke? AB - Positron emission tomography (PET) has been widely used in the study of stroke and related cerebrovascular diseases. It has shown the various stages leading to cerebral infarction and defined the significance of the ischaemic penumbra. PET scan can predict the clinical outcome of patients with acute ischaemic stroke. Several types of diaschisis can also be demonstrated by PET. They reflect different pathophysiological changes in supratentorial infarcts. Post-apoplectic seizures are shown to increase the ischaemic damage in the affected cerebral hemisphere. PET has contributed also to the concept of multi-infarct dementia, although the significance of chronic ischaemia in the pathogenesis of vascular dementia has not been fully investigated. PMID- 9345589 TI - Co-registration of PET and MRI in different courses of MS using Cobalt-55 as a Calcium-tracer. AB - Multiple Sclerosis (MS) is an auto-immune central nervous system (CNS) inflammation. At this moment, MRI is the most accurate paraclinical test in MS to monitor disease activity, although poorly correlated with clinical impairment. PET using Co-55 as a Ca-tracer may visualize Co-transport across the neuronal membrane, Ca-mediated inflammatory processes and passive leakage through a breach in the blood-brain barrier. Co-registration of MRI and Co-PET may actually allow identification of clinically active lesions. MRI and Co-PET were performed as described elsewhere. Based on a statistic parametric mapping (SPM-96)-software package, MRI and Co-PET were superimposed. A semi-automated technique was used to count the MS-lesions. We included four groups of eight MS-patients with relapsing remitting (RR), primary progressive (PP), progressive relapsing (PR) and secondary progressive (SP) courses and eight healthy volunteers. MS was assessed in terms of impairment using Kurtzke's Expanded Disability Status Scale (EDSS) and Scripps Neurological Rating Scale (NRS). Co-PET displayed focal uptake throughout the MS brain, both in the grey and white matter. All four patients groups (as compared to controls) demonstrated a more inhomogeneous distribution of Co-spots with a tendency to show clustering, most evident in RR-MS. SPM analysis revealed an essentially different distribution pattern of MS spots on MRI and Co-PET. (Merging of) Co-PET and MRI may eventually form complementary tools for identifying clinically relevant lesions, thus providing a more reliable secondary outcome measure in MS. PMID- 9345588 TI - 55Co-PET in stroke: relation to bloodflow, oxygen metabolism and gadolinium-MRI. AB - BACKGROUND: Several studies have shown the feasibility of Co-isotopes (55Co and 57Co) in imaging of neuronal damage in stroke, multiple sclerosis, cerebral tumors and trauma. These studies indicate that Co-isotopes allow visualization of brain pathology related to inflammatory processes, reactive gliosis and cell death. Until now, it is not clear if 55Co accumulation occurs in the core of infarction or in the penumbra. Therefore, in the present study, we compared 55Co PET with functional parameters such as cerebral bloodflow (rCBF) using C15O2, oxygen metabolism (rCMRO2) using 15O2 and cerebral bloodvolume (CBV) using C15O in PET and with the anatomical parameter Gd-MRI. PATIENTS AND METHODS: Seventeen patients (11 male; 6 female) age 43 to 84 (mean 69) with middle cerebral artery (mca) stroke, as proven by CT or MRI, were examined with 55Co-PET (0.5-1.0 mCi 55CoCl2), C15O2-, 15O2- and C15O-PET in one session 0-30 days after stroke-onset. Regions of infarction were defined by rCMRO2 being smaller than 65% or rCBF below 45% of the contralateral value and were subsequently superimposed on the cobalt scan. To compare the Cobalt uptake with the Gd-MRI, a realignment program was used that matches the MRI with the bloodflow images. Clinical status was established using the Orgogozo stroke scale at admission and at discharge (at least 6 weeks after admission) and the Barthel index. RESULTS: Eight patients showed a positive Co-PET scan and were used for further analysis. It appeared that Co accumulates in areas with a diminished oxygen metabolism and with a preserved bloodflow. We found Co-uptake in only a part of the Gd enhanced brain tissue with a tendency to be located peripherally or outside the Gd demarcated brain tissue. CONCLUSION: The results of the present study suggest that Co accumulates into infarcted brain tissue with a rather preserved flow independently of blood-brain barrier breakdown. PMID- 9345590 TI - Positron emission tomography and brain tumours. AB - The first applications of positron emission tomography (PET) for the study of brain tumours appeared early in the development of this technology. New trends in these particular PET applications tend to take into account the histological heterogeneity of these tumours and the necessity to integrate PET data in their surgical management. Better knowledge on PET tracers behavior in brain tumours should lead to new clinical uses, in particular in the promising field of cancer treatment evaluation. PMID- 9345591 TI - Positron emission tomography in parkinsonism. AB - Positron emission tomography provides researchers and clinicians with information on cerebral blood flow, metabolic pathways or neurotransmission systems, which may help to understand the physiopathology of movement disorders. Studies of the nigrostriatal dopaminergic pathway and data on cerebral glucose metabolism contribute to discriminate between parkinsonian syndromes. New techniques for image processing and statistical analysis, and brain activation studies better emphasize dysfunction of cortico-subcortical neuronal networks responsible for movement disorders, or functional modification after therapeutic action. PMID- 9345592 TI - Positron emission tomography in dementia. AB - Positron emission tomography (PET) studies in dementia suffer from methodological differences. Most PET studies concern resting state metabolism in Alzheimer's disease and its correlations with neuropathology and neuropsychology. Only few clinically based PET studies on sensitivity and specificity of specific findings in specific dementing disorders have been reported. The role of PET in dementia for routine clinical purposes seems limited at present. Future studies should evaluate the potential of PET in preclinical diagnosis and to study treatment effects. PMID- 9345593 TI - PET studies in epilepsy. AB - Positron emission tomography (PET) plays a major role in the pre-surgical evaluation of patients with refractory partial epilepsy. In this review we discuss the limitations of PET data, the possibilities of fluorodeoxyglucose PET studies, neuroreceptor PET studies and alternatives, such as SPECT and MRI. PMID- 9345595 TI - Nutritional significance of phytic acid and phytase. AB - In the nutrition of monogastric animals phytate-P represents a poorly available source of phosphorus, especially in the case of diets low in phytase activity. Similarly the bioavailability of different minerals and trace elements is considerably reduced by phytate complexes. High concentrations of Ca increase the anti-nutritive effect of phytic acid on mineral and trace element bioavailability and thus impede the action of phytase. This effect can in part be compensated by an increased supply of vitamin D. There is also evidence for protective functions of phytic acid such as the prevention of the formation of free radicals, the delaying of post prandial glucose absorption, the decrease in plasma cholesterol and triglycerides as well as a change in the carry over of heavy metals. The basic mechanisms by which phytic acid may exert these effects are still not clear. In several studies reported in the literature, evidence for the nutritional significance and ecological importance of microbial phytase for pigs and poultry has been given. As the monogastric organism contains no or only negligible amounts of endogenous phytase in the stomach and small intestine, it is therefore dependent on plant or microbial phytase. Plant phytase, e.g. from rye, triticale, wheat or, in smaller amounts from barley, and supplemented Aspergillus-phytase display cumulative effects. PMID- 9345594 TI - Imaging of receptors in clinical neurosciences. AB - This article deals with the question why should one determine receptors in the brain with positron and single photon emission tomography (PET and SPECT, respectively). Radiopharmaceuticals for a wide variety of receptors are available now. Receptors studies with PET and SPECT have thus far focused on the following issues: occupancy during drug treatment, quantification in neuropsychiatric diseases and visualizing specific pathology. Far most studies on receptor occupancy are concerned with antipsychotic treatment in schizophrenic patients. During treatment with classical antipsychotic drugs more than 65% of the receptors are occupied in both responding and drug therapy resistant patients. High occupancy appeared to be linked to frequent occurrence of extrapyramidal symptoms (parkinsonism). Atypical antipsychotic drugs, producing no parkinsonism, are effective even when less than 50% dopamine D2 receptors are blocked. Quantification of receptor density in vivo depends very much on the mathematical models used, and is often very difficult and thus far of little clinical use. Finally, receptors can be used as markers for specific neurons or other cells. In the caudate nucleus dopamine receptors are localized in neurons that degenerate in Huntington's disease, so early in the disease decreases in receptor binding can be shown. In Alzheimer disease decreases in neural receptor and increases in omega-3 receptors are observed indicating degeneration and inflammation, respectively. PMID- 9345596 TI - Transformation of nivalenol by gastrointestinal microbes. AB - The capacity of the gastrointestinal microflora of pig, cow, and chicken to metabolize nivalenol (NIV) and deoxynivalenol (DON) was studied both in vivo and in vitro. Before feeding NIV to pigs, no metabolites of NIV or DON were formed in anaerobic incubates of the toxins with the pigs feces. However, after one week on a diet containing 2.5 or 5 ppm NIV, nearly all excreted NIV in feces had been de epoxidated in five of six pigs. After three weeks on the NIV diet also the sixth pig had acquired this ability. Deoxynivalenol was also de-epoxidated when incubated in vitro with the microorganisms that formed de-epoxy-NIV in vivo. Anaerobic incubation of NIV and DON with cow rumen fluid produced de-epoxides of both toxins in a high proportion. No de-epoxide of NIV, but another unidentified metabolite was found in feces from chicken fed 2.5 or 5 ppm NIV for three weeks. PMID- 9345597 TI - Non starch polysaccharide hydrolyzing enzymes as feed additives: detection of enzyme activities and problems encountered with quantitative determination in complex samples. AB - Chromogenic substrates, an agar diffusion assay and viscosity reduction were used to estimate beta-glucanase and xylanase activities in water soluble extracts of different feedstuffs and digesta supernatants. The dinitrosalicylic acid reducing sugar method was employed to calibrate results from different methods based on international units (IU, glucose equivalents). The detection of dye release from chromogenic substrates was a suitable method, allowing the detection of 0.05 IU of enzyme activity per ml of extract, although measurements in digesta supernatants were limited in linearity (0.1-0.5 IU/ml supernatant). With the agar diffusion assay the detection of enzyme activity was possible over a wider concentration range (extracts: 0.05-1 IU/ml, digesta supernatants: 0.1-1 IU/ml), but visual evaluation led to inaccurate measurement. Accuracy can be improved by computer based evaluation of digital images. The use of viscosity reduction produced linear standard curves from 0.01 to 0.5 IU/ml in feed extracts, but reliability of measurements depended on modification of substrates. Quantification of enzyme activities was influenced by matrix effects of complex samples. Cereal dependant differences were found in various extracts of feed mixtures and cereal extracts. Digesta supernatants partly inhibited enzyme activity, depending on the origin of the sample. Interaction of substrates with digesta components varied between methods. The sensitivity of the methods is comparable, however, all methods require specific calibrations to account for matrix- and enzyme specific effects. PMID- 9345598 TI - Protein degradability, amino acid composition, trypsin inhibitor and urease activity of raw and heat-treated fullfat soybean. AB - The effects of different heat treatments were studied on chemical composition, protein degradability, amino acid composition, trypsin inhibition and urease activity. Three lactating Holstein cows fitted with rumen cannulae were used. Fullfat soybean was prepared employing different forms of heat treatment: dry extrusion at 150 degrees C for 25 s (treatment I); wet-extrusion at 95 degrees C for 30 min (treatment II); toasted soybean at 105 degrees C for 30 min (treatment III) extracted soybean meal (treatment IV); and untreated soybean (treatment V). The incubation times were 0, 2, 4, 8, 16, 24 and 48 h. Samples of raw and heat treated soybean before incubation and the undegraded fraction after 4 and 16 h of incubation were analyzed for amino acids. The results showed that heat treatments did not modify chemical composition, but significantly reduced the content of trypsin inhibition and urease activity, as well as decreased protein degradability. The dry extrusion technique was comparatively the most effective. Amino acid content was not significantly influenced by different techniques, but the quantity of amino acids escaping degradation in the rumen increased. PMID- 9345601 TI - Research of psychosocial issues of children with craniofacial anomalies: progress and challenges. PMID- 9345599 TI - Antisense insulin-like growth factor I transferred into a rat hepatoma cell line inhibits tumorigenesis by modulating major histocompatibility complex I cell surface expression. AB - We have established a hepatocarcinoma cell line (LFCI2 A) that produces voluminous tumors when injected subcutaneously into syngeneic Commentry rats. These neoplastic cells express both insulin-like growth factors (IGF) I and II. When transfected with an episomal cassette-expressing IGF-I antisense RNA, the modified LFCI2 A cell lines become poorly tumorigenic and, when injected subcutaneously, are associated with inhibition of the growth of the parental tumoral cells and/or induction of regression of established tumors. By contrast, cell lines isolated after transfection with the IGF-II antisense-expressing vector were as tumorigenic as the parental cell lines. The results are discussed in terms of protective immunity induced by the tumoral cells transfected by the IGF-I antisense vector. In the transfected hepatocarcinoma cells that do not produce IGF-I, the expression of major histocompatibility complex class I antigen was increased at least 4-fold compared with parental cells. The introduction of these cells in vivo induced a tumor-specific immunity that was associated with CD8 T cells. PMID- 9345600 TI - Plasmovirus: replication cycle of a novel nonviral/viral vector for gene transfer. AB - We recently described a novel nonviral/viral vector for gene transfer, the plasmovirus (Noguiez-Hellin P, Robert-le Meur M, Laune S, et al. C R Acad Sci Paris, Sciences de la Vie. 1996;319:45-50; Noguiez-Hellin P, Robert-le Meur M, Salzmann J-L, et al. Proc Natl Acad Sci USA. 1996;93:4175-4180). Plasmoviruses are plasmids capable of expressing all the viral genes required for generating infectious particles and packaging a defective genome containing a transgene. Transfected as plasmids, plasmoviruses transform the transduced cells into packaging cells that release infectious replication-defective retrovirus vectors (RV) containing a transgene, which are capable of infecting nearby cells. We previously showed that such a vector can efficiently "propagate" the transgene after transfection. Here we examine in greater detail the different steps of plasmovirus replication in vitro in human (143 B TK-) and murine (NIH 3T3 TK-) cells. Molecular-biological analysis revealed plasmovirus-coded protein expression starting from 24 hours post-transfection, followed by the detection of infectious RV 48 hours post-transfection. The gag proteins were correctly processed in the released particles. Electron microscopic analysis revealed typical type C particles. Nonintegrated plasmovirus DNA was not toxic for the cells and could be detected for at least 14 days post-transfection. While the transfected gag gene and the transgene could also be detected throughout this period, we observed that env-coded proteins decreased after 72 hours post transfection. Nevertheless, the production of RV resulted in the propagation of the transgene in the culture, with stable integration of plasmovirus proviral DNA into the host genome of infected cells. We show that this propagation results in a major improvement in therapeutic efficacy using an HSV1-TK transgene and ganciclovir treatment, when compared to that of plasmovirus constructs that cannot propagate. Altogether, these results demonstrate the functionality of this gene transfer method and suggest that improvements in the vector design enhance its efficacy. PMID- 9345602 TI - Presurgical and postsurgical mental and psychomotor development of infants with sagittal synostosis. AB - OBJECTIVE: The current study compared the mental and psychomotor development of infants with nonsyndromic sagittal synostosis (SS) with a demographically matched comparison group without congenital defects. Within the SS group, we tested the hypothesis that age of cranial release would be inversely correlated with mental development. DESIGN: The design was prospective and longitudinal. Participants were assessed at 4, 12, and 24 months of age. SETTING: The study was conducted in a craniofacial clinic at an urban children's hospital. PARTICIPANTS: Participants were 19 infants with SS (consecutive craniofacial program referrals) and 19 demographically matched comparison infants recruited from the community. One infant with SS did not attend the 24-month assessment. MAIN OUTCOME MEASURES: Mental and Psychomotor Indices from the Bayley Scales of Infant Development were the primary outcome measures. Subdomains of development were created using Kohen Raz scoring procedures. All measures were determined a priori. RESULTS: Repeated measures MANOVAs revealed no statistically significant differences in the developmental trajectories of the two groups. None of the SS group infants received Mental Development Index (MDI) scores in the mentally retarded or borderline range of intellectual functioning (i.e., below 78). An inverse correlation (r = -.30) was found between the age at surgery and Bayley growth curve coefficients; however, this association was not statistically significant (p = .10, one-tailed). CONCLUSIONS: Results are consistent with previous studies of the mental and psychomotor development of infants with nonsyndromic craniosynostoses in relation to normative test data. The relation between surgery age and developmental outcome merits further study in a larger sample with a greater range of surgery ages. PMID- 9345603 TI - Observed social interaction patterns in adolescents with and without craniofacial conditions. AB - OBJECTIVE: This study examined social interactions of adolescents in a natural environment (school lunch room) to determine if there were identifiable differences in social behavior between children with and without craniofacial conditions (CFC). DESIGN: This was an observational study comparing social interaction skills of children with CFC to peers without craniofacial conditions. SETTING: The observations were conducted in the respective school lunch rooms of the adolescents with CFC. PARTICIPANTS: Clinical subjects were 13 adolescents (4 male) with various craniofacial conditions (5 cleft lip and palate) and 12 (4 male) peers without CFC present in the same lunch room. MAIN OUTCOME MEASURES: An unknown observer obtained 45 minutes of structured observational data on subject initiations, responses, nondirected comments, and extended conversations over two to three lunch room periods. Data was coded on the Epson HX-20 for type, frequency, and duration of social contact. Specific measures included: subject initiations and responses, peer initiations and responses, conversations events, and nondirected comments. RESULTS: Statistically significant differences were found between CFC and comparison subjects (CS) on each social interaction variable measured. CS initiated more contacts, received positive responses more frequently, and engaged in longer conversations than CFC subjects ([F (1,24) = 14.1, p < .01; F (1,24) = 67.2, p < .001; F (1,24) = 5.50, p < .05]. CS were approached by and responded appropriately to peers more often [F (1,24) = 28.1, p < .001; F (1,24) = 43.2, p < .001]. Subjects with CFC were more likely to produce nondirected comments (N = 7, x = 0, p < .01). CONCLUSIONS: A significant number of children with CFC behaved differently than their peers in a natural, daily occurring situation. They were often at the periphery of the group, observers rather than participants in conversation. PMID- 9345604 TI - Personality attributions based on speech samples of children with repaired cleft palates. AB - OBJECTIVE: This study examined whether or not assumptions made about personality characteristics based on speech samples differed for children with repaired cleft palates (CP) versus unaffected children. DESIGN: Audiotapes of speech samples were presented in random order to blind raters. PATIENTS/PARTICIPANTS: The subjects were 20 children (10 females, 10 males) with repaired CP and 16 control (i.e., unaffected) children (8 females, 8 males). All children were 8 to 12 years of age, Caucasian, living in the St. Louis area, and lower-middle to upper-middle class. The raters were 20 (13 females, 7 males) 6th grade Caucasian students who attend a private school in the area. SETTING: Raters heard tapes in a group setting, but with individual headphones, in their school's cafeteria. MAIN OUTCOME MEASURE: Each speech sample was rated (7-point Likert scale) by each student rater on a variety of personality characteristics based on the "Big Five" personality factors. RESULTS: A factor analysis of the items revealed a two factor solution, although the factors were highly negatively correlated. No significant differences were found between ratings for the CP sample and the control sample for either factor scale (ANOVA, p = .93; p = .67). Similarly, when the two factors were combined to form a single factor, no significant differences were found between the ratings for the CP sample and the control sample (ANOVA, p = .79). CONCLUSIONS: Overall, it does not appear that children differentially associated personality characteristics based on speech to children with repaired CP versus unaffected children, in the absence of visual input. PMID- 9345605 TI - Facial and speech relationships to behavior of children with clefts across three age levels. AB - OBJECTIVE: This study was conducted to examine relationships of facial and speech ratings to behavior at three ages. DESIGN: Multiple correlations to control for gender, IQ, and socioeconomic status (SES). Partial correlations between speech or facial ratings and behavior. SETTING: A university hospital interdisciplinary cleft/craniofacial center. PATIENTS: Sixty-five children with CP or CLP and no other anomalies seen at ages 6, 9, and 12 years. INTERVENTIONS: Speech ratings, facial ratings, hearing assessment, and SES ratings, were collected at ages 6, 9, and 12. MAIN OUTCOME MEASURE: The Behavior Problem Checklist was used and internalizing problems were expected. RESULTS: Less severe speech problems associated with behavioral inhibition were noted at age 9 years (p < .01). Greater facial disfigurement was correlated with greater inhibition at age 12 (p < .001). CONCLUSIONS: Longitudinal data indicates different associations of speech and facial variables were made to behavior at different ages (9 and 12). No significant relationships were identified at age 6. PMID- 9345606 TI - Self-perceived facial appearance and psychosocial adjustment in preadolescents with craniofacial anomalies. AB - OBJECTIVE: To identify aspects of psychosocial adjustment related to the self perceived facial appearance of preadolescents with craniofacial abnormalities. DESIGN: Concurrent relationships were evaluated using a within-group correlational design. PARTICIPANTS: Participants were 24 patients, aged 11 to 13, of a major craniofacial center and their parents who were contacted by telephone and agreed to participate. MAIN OUTCOME MEASURES: Self-report and parent-report questionnaires assessing psychosocial adjustment. RESULTS: Self-perceived facial appearance was positively correlated with global self-worth, self-perceived social acceptance, and number of same-sex close friends, and negatively correlated with loneliness, parent-rated social problems, and parental advice/support and concern (all p's < .05 or better). CONCLUSIONS: Dissatisfaction with facial appearance was associated with peer relationship problems and low global self-esteem, but not with other aspects of self-concept or other types of adjustment problems. PMID- 9345607 TI - Psychological research of children with craniofacial anomalies: review, critique, and implications for the future. PMID- 9345608 TI - Comparison of nasopharyngeal growth between patients with clefts and noncleft controls. AB - OBJECTIVE: This study was a comparison of the cephalometric growth characteristics of the nasopharyngeal structures between UCLP and noncleft controls. METHOD: Eighty patients with complete unilateral cleft lip and palate (UCLP group) and 82 noncleft controls (NCC group) were assigned to four developmental stages (i.e., stage 1, at 4 years; stage 2, at 8 years; stage 3, at 12 years; and stage 4, at 17 years of age). Measurements on the anteroposterior and the vertical dimensions were derived from reference lines and points of nasopharyngeal structures on the lateral cephalograms. RESULTS: The results showed that there were no growth differences between the two groups at any stages in the regions of cranial base and cervical vertebrae, and that growth of the posterior maxilla in the UCLP group was significantly less at any stage in both A P and vertical dimensions than in the NCC groups. As well, the nasopharyngeal triangle (Ho-At-PMP) in the groups showed almost parallel increase with stage, though with short vertical dimension in the UCLP group, and the soft palate length in the UCLP group was significantly less at stages 2, 3, and 4 compared to that in the NCC group. The adequate ratio (soft palate length/pharyngeal depth) in the UCLP group tended to decrease and was significantly less at stage 4 compared to that in the NCC group. CONCLUSIONS: These results indicate that the growth of the cranial base and the upper cervical vertebrae is independent of the effect of clefts or of surgeries on clefts, and that the growth inhibition at the posterior maxilla results in morphologic disharmony of upper nasopharyngeal structures. This could be a potential factor for the reappearance of velopharyngeal incompetence at a later age. PMID- 9345609 TI - Mandibular asymmetry in noncleft and unilateral cleft lip and palate individuals. AB - OBJECTIVE: The purpose of this study was to retrospectively investigate mandibular asymmetry in unilateral cleft lip and palate individuals (UCLP) in relation to chronologic age and in relation to lower facial asymmetry. DESIGN: The longitudinal records of 34 UCLP individuals and 142 controls treated in the Department of Orthodontics, Eastman Dental Center, Rochester, NY, were included in the study. Posteroanterior and oblique cephalometric radiographs were analyzed for lower facial asymmetry and mandibular asymmetry, respectively. Mandibular asymmetry in UCLP was analyzed relative to three age groups (6-10, 11-14, and 15 or greater) and compared to controls. Moreover, mandibular asymmetry was analyzed relative to lower facial asymmetry. RESULTS: UCLP individuals showed no significant differences in mandibular asymmetry compared to controls. In addition, no significant correlation was found between mandibular asymmetry and lower facial asymmetry in UCLP. CONCLUSIONS: The degree of mandibular asymmetry in UCLP appears not to be the major contributing factor to the lower facial asymmetry noted on these individuals. Possible cranial-base/temporal-region anomalies may be involved in unilateral cleft lip and palate and be responsible of the asymmetry noted in the lower facial skeleton. PMID- 9345610 TI - Craniofacial morphology in patients with Kallmann's syndrome with and without cleft lip and palate. AB - OBJECTIVE: Kallmann's syndrome is characterized by the association of hypogonadotropic hypogonadism and anosmia or hyposmia. The principal endocrine defect of hypogonadotropic hypogonadism is a failure to secrete luteinizing hormone-releasing hormone (LHRH), resulting in underdevelopment of the pituitary gonadotropes and an inability to synthesize and release luteinizing hormone and follicle-stimulating hormone. The purpose of the present investigation was to describe the dentition and the craniofacial morphology in patients diagnosed with Kallmann's syndrome. DESIGN: The sample consisted of 11 patients, 2 of whom also had bilateral cleft lip and palate. Radiographic investigations, including cephalometry, were performed. Comparisons were made to normal individuals and to cleft lip individuals without Kallmann's syndrome. RESULTS: Dentition: tooth agenesis occurred more frequently in patients with Kallmann's syndrome. Craniofacial morphology: Increased mandibular inclination and mandibular angulation were seen in Kallmann patients. When clefting also occurred, extreme retrognathism of both maxilla and mandible was seen, a deviation which seemingly worsened during growth. The anterior cranial base and the sphenoid bone showed an altered morphology in one of the patients with Kallman's syndrome. CONCLUSIONS: An early diagnosis of Kallmann's syndrome is very important because the prognosis for endocrine treatment thereby improves, and therefore, it is recommended that the sense of smell be evaluated in patients with the craniofacial morphology described. PMID- 9345611 TI - Craniofacial morphology of conotruncal anomaly face syndrome. AB - OBJECTIVE AND DESIGN: The conotruncal anomaly face syndrome (CTAF) comprises congenital heart disease and dysmorphic face, and is frequently associated with cleft palate or hypernasality. There have been many discussions about the overlap with velocardiofacial syndrome (VCF). The aim of this study was to clarify the craniofacial characteristics of CTAF patients by clinical examination, and photogrammetric and cephalometric analyses, and to clarify the differences compared to published data on VCF. RESULTS: The facial features of CTAF included hypertelorism, small palpebral fissures, upward slanting of palpebral fissures, bloated eye lids, low nasal bridge, small mouth, open mouth at rest, and malformed auricles. Cephalometric features included bialveolar protrusion, small gonial angle, backward rotation of the mandibular ramus, and labial inclination of the maxillary incisors. An acute cranial base angle was also noted. These results differed from those of VCF. There were, however, no obvious pathognomonic findings for the differential diagnosis between CTAF and VCF. CONCLUSIONS: Considering these findings, use of CATCH 22, the inclusive classification of cardiac anomalies, cleft palate, and dysmorphic face may be of value for the clinical understanding in these patients. PMID- 9345612 TI - Comparative survey of osteotomized and nonosteotomized BCLP patients. AB - OBJECTIVE: At Hannover Medical School, treatment of BCLP patients was revised and updated in 1980. The objective of the present study was to evaluate the differences in treatment outcome between BCLP patients treated after the revised concept including infant orthopedics, and BCLP patients who received osteotomy in addition to surgical and orthodontic treatment during childhood. PATIENTS: Nine of 48 BCLP patients born between 1980 and 1983 received surgical and orthodontic treatment according to the Hannover concept. They were compared to 9 of 68 adolescent and adult patients from Hannover without this protocol, who underwent maxillary osteotomy and consecutive orthodontic treatment. MAIN OUTCOME MEASURES: Comparison of the two groups was made at the end of active orthodontic treatment by cast analysis and lateral cephalometrics to evaluate sagittal, transverse, and vertical changes. RESULTS AND CONCLUSIONS: No patient treated using the revised protocol showed characteristics of skeletal angle class III at any stage of investigation. No indication for osteotomy was found in this group. All patients with osteotomy had skeletal angle class III resulting from insufficient midfacial growth. Sagittal and vertical skeletal relations were successfully improved by osteotomy. PMID- 9345613 TI - Sagittal position of the left and right maxillary segment in children with cleft lip and palate. AB - OBJECTIVE: The purpose of the present study was to investigate left-right differences in the sagittal position of the maxillary segments in children with cleft lip and palate. METHOD: The sample consisted of children with operated cleft lip or cleft lip and alveolus [CL/CLA (n = 16) mean age, 9.3 yr], operated unilateral cleft lip and palate [UCLP (n = 27) mean age, 9.1 yr], and operated bilateral cleft lip and palate [BCLP (n = 17) mean age, 9.5 yr]. Computed tomography (CT) horizontal slices of the maxilla were obtained and used to determine the sagittal position of the left and right segment of the maxilla in relation to the mandibular rami and the cranial base. Significant effects were analyzed with multivariate analyses of variance (MANOVA). RESULTS AND CONCLUSIONS: It was concluded that, in contrast to children having CL/CLA or UCLP, children with BCLP showed left-right differences in the sagittal position of the maxillary segments. The segment on the left side was more posteriorly positioned compared to the right side. Because the same results were obtained in relation to the mandibular rami as well as in relation to the cranial base, it can be assumed that the position of these rami are not affected by the different types of oral clefts. PMID- 9345614 TI - Continuity of care: University of Iowa Cleft Lip/Palate Interdisciplinary Team. AB - INTRODUCTION: Patient satisfaction with the continuity of care is an important aspect of care delivery that may be evaluated to assess team effectiveness. This study was conducted to evaluate the number of patients seen by each team member, and the patients' perceived importance of continued care by the same health care provider during their overall treatment. METHODS: A survey was constructed by the members of the Cleft Lip/Palate Team at the University of Iowa Hospitals and Clinics. One hundred and thirty-eight subjects were invited to participate during consecutive clinic days over a 5-month period; 101 subjects (73.1%) responded. RESULTS: The survey revealed that the percentage of patients seen by each member of the team ranged from 92.1% (93/101) for the surgeon to 24.8% (25/101) for the genetic counselor. Strong agreement with continuity of care by health professionals ranged from 85.6% (83/97) for the surgeon, 63.7% (58/91) for the orthodontist, 63% (17/27) for the psychologist, to 23.1% (19/82) for the audiologist. The percentages dropped somewhat if the patients thought that they may have to wait longer for their next appointment. CONCLUSION: There is a strong preference for continuity of health care. PMID- 9345616 TI - Effect of alveolar bone grafting in the mixed dentition on maxillary growth in complete unilateral cleft lip and palate patients. AB - OBJECTIVE: This study was conducted to evaluate the effects on facial growth of alveolar bone grafting in the mixed dentition for patients with UCLP. DESIGN: Retrospective cephalometric study. SETTING: Craniofacial Treatment and Research Centre, The Hospital for Sick Children, Toronto, Ontario, Canada. PATIENTS: The 58 patients participating in the study had a history of complete unilateral cleft lip and palate, all of which were repaired by the same plastic surgeon. INTERVENTIONS: Twenty-one patients received an iliac-crest alveolar bone graft at a mean age of 10.3 years, while 37 did not receive an alveolar bone graft. Lateral cephalometric radiographs were obtained on all patients at two different times:at a mean age of 9.4 years (prior to bone grafting in the grafted group) and at a mean age of 15.2 years. MAIN OUTCOME MEASURES: All radiographs were traced and digitized by the same person, using cephalometric computer software. Superimposition and cephalometric analysis was undertaken to investigate the differences between the two groups in the 5.6-year experimental period. A two-way analysis of covariance was used for evaluation of the statistical significance of the results. RESULTS: No statistically significant differences were found in 14 of the 15 cephalometric measurements performed. Harvold's maxillary unit length was statistically significantly shorter in the grafted group, although a lack of correlation with angular measurements and inherent problems with this specific measurement raise doubts in this finding. CONCLUSION: Mixed dentition bone grafting does not affect subsequent vertical and A-P development of the maxilla in complete unilateral cleft lip and palate patients during the first several postoperative years. PMID- 9345615 TI - Testing for interaction between maternal smoking and TGFA genotype among oral cleft cases born in Maryland 1992-1996. AB - OBJECTIVE: Infants born in Maryland between June 1992 and June 1996 were used in a case-control study of nonsyndromic oral clefts to test for effects of maternal smoking and a polymorphic genetic marker at the transforming growth factor alpha (TGFA) locus, both of which have been reported to be risk factors for these common birth defects. DESIGN AND SETTING: Cases were infants with an oral cleft ascertained through three comprehensive treatment centers, with additional ascertainment through a registry of birth defects maintained by the Maryland Health Department. Controls were healthy infants. Medical history information on infants and mothers were collected, along with DNA samples. PATIENTS, PARTICIPANTS: Among 286 cases contacted (72% ascertainment), there were 192 nonsyndromic isolated oral clefts (106 M; 86 F) available for this case-control study. MAIN OUTCOME MEASURES: The largest group of 149 Caucasian nonsyndromic cases and 86 controls was used to test for association with maternal smoking and genotype at the Taq1 polymorphism in TGFA. RESULTS: While this modest sample had limited statistical power to detect gene-environment interaction, there was a significant marginal increase in risk of having an oral cleft if the mother smoked (odds ratio = 1.75, 95% CI = 1.01 to 3.02). We could not demonstrate statistical interaction between maternal smoking and TGFA genotype in this study, however, and the observed increase in the C2 allele among cases was not statistically significant. CONCLUSIONS: We could not confirm either the reported association between oral clefts and TGFA genotype or its interaction with maternal smoking. However, these data do show an increased risk if the mother smoked during pregnancy, and this effect was greatest among infants with a bilateral cleft and no close family history of clefts. PMID- 9345617 TI - Carbohydrates and glycoconjugates. PMID- 9345618 TI - Oligosaccharide structures: theory versus experiment. AB - Recently, the interdependency of theoretical and experimental approaches in the structure determination of oligosaccharides has been confirmed. More accurate simulations are possible because of the advances in software and computers. Meanwhile, improvements in NMR techniques permit the measurement of numerous structural and dynamical parameters, either for the free state or for carbohydrate ligands bound to receptors. Several crystal structures of isolated or protein-complexed oligosaccharides give new clues for modeling the intermolecular forces that drive the interactions. PMID- 9345619 TI - Cell-surface carbohydrate recognition by animal and viral lectins. AB - Many animal and viral lectins are specific for monosaccharides found in particular glycosidic linkages, or for larger oligosaccharide structures. Recent crystal structures of complexes between these proteins and receptor fragments have provided insights into the recognition of linkage isomers and oligosaccharide conformation. PMID- 9345620 TI - New folds of plant lectins. AB - Among the crystal structures of lectins determined recently, three--snowdrop lectin, jacalin and amaranthin--represent new lectin families. Their polypeptide folds share remarkably similar features and consist exclusively of beta structure. Autonomously folded beta-sheet subdomains, inter-related by a pseudothreefold symmetry, assemble to form beta-prism or beta-barrel structures which are stabilized by a hydrophobic core. PMID- 9345621 TI - Structural and sequence-based classification of glycoside hydrolases. AB - The diversity of oligo- and polysaccharides provides an abundance of biological roles for these carbohydrates. The enzymes hydrolysing these compounds, the glycoside hydrolases, therefore mediate a wealth of biological functions. Glycoside hydrolases fall into a number of sequence-based families. The recent analysis of these families, coupled with the burgeoning number of 3D structures, provides a detailed insight into the structure, function and catalytic mechanism of these enzymes. PMID- 9345622 TI - Mechanism of catalysis by retaining beta-glycosyl hydrolases. AB - Recent structural studies provide a fresh look at the catalytic mechanism of polysaccharide hydrolysis by retaining beta-glycosyl hydrolases. Highlights include insights into saccharide ring distortion, both upon binding and during the course of catalysis, and evidence for the regulation of the pKa of key catalytic residues. PMID- 9345623 TI - Enzyme-catalyzed formation of glycosidic linkages. AB - Significant progress has recently been achieved in the use of glycosidases and glycosyltransferases as synthetic tools. Glycosidases have been used to synthesize trisaccharides with a reasonable overall yield, as well as high mannose neoglycoconjugates. Studies on glycosyltransferases have defined reaction mechanisms and demonstrated reasonable substrate tolerance of these enzymes. Effective methodology for the synthesis of defined glycoproteins has also been demonstrated. PMID- 9345624 TI - Biological significance of lipid polymorphism: the cubic phases. PMID- 9345626 TI - Phasing methods for protein crystallography. AB - The phase problem has been formulated as one of constrained global minimization and it leads to the minimal principle, which is the theoretical basis of 'Shake and-Bake'--an algorithm for the automatic solution of the phase problem, ab initio. A related approach (termed 'Half-Baked') that, as in Shake-and-Bake, alternates phase refinement in reciprocal space with density modification in real space, has recently been formulated. The traditional techniques of direct methods have been integrated with isomorphous replacement and anomalous scattering and the first applications have been made. The accurate measurement of a limited number of structure invariants by means of multiple beam X-ray diffraction has become a reality. A feasibility study shows that, in combination with the direct method, the ability to measure the values of the structure invariants experimentally will strengthen existing techniques. PMID- 9345627 TI - The benefits of atomic resolution. AB - After a long gestation, the elucidation of the crystal structures of proteins at atomic resolution is now maturing. The use of such data for both refinement and structure solution is advancing space. The necessary technology is generally available, in terms of data collection, computing hardware and software. The structures appearing in the literature mainly relate to demonstration projects on native proteins. The importance of these alone is already obvious. Biologically significant results, in terms of ligand complexes and prosthetic groups, are just starting to emerge. PMID- 9345628 TI - Synchrotron radiation applications to macromolecular crystallography. AB - Progress has been rapid in the development and application of four different types of macromolecular crystallographic experiment at synchrotron hard X-ray sources: multiwavelength anomalous diffraction; studies of crystals with very large unit cell dimensions; structure determination at atomic or near-atomic resolution; and time-resolved studies. The results illustrate the interplay between the advanced technical capabilities available at new beamlines and more challenging scientific issues. PMID- 9345629 TI - Crystallization of membrane proteins. AB - Five new membrane protein structures have been determined since 1995 using X-ray crystallography: bacterial light-harvesting complex; bacterial and mitochondrial cytochrome c oxidases; mitochondrial bc1 complex; and alpha-hemolysin. These successes are partly based on advances in the crystallization procedures for integral membrane proteins. Variation of the size of the detergent micelle and/or increasing the size of the polar surface of the membrane protein is the most important route to well-ordered membrane protein crystals. The use of bicontinuous lipidic cubic phases also appears to be promising. PMID- 9345630 TI - Insights into biomolecular function from small-angle scattering. AB - Recent advances in neutron and X-ray sources and instrumentation, new and improved scattering techniques, and molecular biology techniques, which have permitted facile preparation of samples, have each led to new opportunities in using small-angle scattering to study the conformations and interactions of biological macromolecules in solution as a function of their properties. For example, new instrumentation on synchrotron sources has facilitated time-resolved studies that yield insights into protein folding. More powerful neutron sources, combined with molecular biology tools that isotopically label samples, have facilitated studies of biomolecular interactions, including those involving active enzymes. PMID- 9345632 TI - Time-resolved mid-infrared spectroscopy: methods and biological applications. AB - Recent developments in time-resolved infrared spectroscopy have paved the way to probe transient intermediates with a high degree of functional group specificity on timescales as short as femtoseconds. This capability has been exploited in studies of biophysical phenomena ranging from protein folding/unfolding to ligand migration in proteins. PMID- 9345631 TI - Scanning force microscopy under aqueous solutions. AB - Merely ten years after its invention, the scanning force microscope is becoming a powerful method to investigate the structure and dynamics of biological molecules under aqueous environments. From the visualization of transcription in real time to the mechanical manipulation of individual proteins, the advances made during the past year open up a vast number of exciting applications of this technique in biology. PMID- 9345633 TI - Solution NMR spectroscopy beyond 25 kDa. AB - Improvements in NMR instrumentation, higher magnetic field strengths, novel NMR experiments and new deuterium-labeling strategies have significantly increased the scope of structural problems that can now be addressed by solution NMR methods. To date, a number of structures of proteins of 30 kDa have been solved using multidimensional 15N,13C,2H NMR techniques, and this molecular weight limit will probably be surpassed in the near future. PMID- 9345634 TI - Probing molecular motion by NMR. AB - Recently developed solution NMR methods for measuring 2H, 13C, and 15N spin relaxation, coupled with biosynthetic isotopic enrichment, permit the characterization of backbone and sidechain dynamical properties of proteins on picosecond/nanosecond and microsecond/millisecond timescales. Theoretical interpretations of the relaxation data provide insights into the biophysical and functional properties of proteins. PMID- 9345636 TI - Carbohydrates and glycoconjugates. PMID- 9345635 TI - Femtosecond spectroscopy of photosynthetic light-harvesting systems. AB - Observing the elementary steps of light-harvesting in real time is now possible using femtosecond spectroscopy. This, combined with new structural data, has allowed a fairly complete description of light-harvesting in purple bacteria and substantial insights into higher plant antenna systems. PMID- 9345637 TI - Biophysical methods. PMID- 9345638 TI - Insulin action on metabolism. PMID- 9345639 TI - Insulin resistance. PMID- 9345641 TI - Fatty acids and beta-cell function. PMID- 9345640 TI - The adipose tissue/central nervous system axis. PMID- 9345642 TI - Beta-cell ontogeny: growth and death. PMID- 9345643 TI - Insulin production: from gene to granule. PMID- 9345644 TI - Surrogate beta cells. PMID- 9345645 TI - Beta-cell replacement. PMID- 9345646 TI - Immunology of IDDM. PMID- 9345647 TI - Prediction and prevention of IDDM. PMID- 9345648 TI - Epidemiology and genetics of diabetic complications. PMID- 9345649 TI - Pathophysiology of diabetic complications. PMID- 9345650 TI - Growth factors and diabetic nephropathy: kidney structure and therapeutic interventions. PMID- 9345651 TI - Diabetic neuropathies. PMID- 9345653 TI - Hypoglycaemia and glucose sensing. PMID- 9345652 TI - Cardiovascular complications of diabetes. PMID- 9345654 TI - New insulins and other possible therapeutic approaches. PMID- 9345655 TI - Defining safe drinking water. PMID- 9345656 TI - Defining and explaining race effects. PMID- 9345657 TI - A new insight into pregnancy loss. PMID- 9345658 TI - Condoms and urinary tract infections: is nonoxynol-9 the problem or the solution? PMID- 9345659 TI - Drinking water turbidity and pediatric hospital use for gastrointestinal illness in Philadelphia. AB - Recent outbreaks have demonstrated that serious infectious gastrointestinal illness related to drinking water supplies remains a problem in the United States. The magnitude is unknown, but children, the elderly, and immunocompromised individuals are considered at highest risk. We examined the association between daily measures of drinking water turbidity and both emergency visits and admissions to Children's Hospital of Philadelphia for gastrointestinal illness, controlling for time trends, seasonal patterns, and temperature. We found that an interquartile range increase in turbidity levels in Philadelphia drinking water was associated with a 9.9% increase [95% confidence limits (CL) = 2.9%, 17.3%] in gastrointestinal emergency visits for children age 3 years and older 4 days later. For children age 2 years and younger, an association was found with a lag of 10 days (5.9% increase; 95% CL = 0.2, 12.0). For admissions, a similar pattern was seen. For children over 2 years old, an increase of 31.1% (95% CL = 10.8%, 55%) was seen with a lag of 5-6 days. For younger children, an increase of 13.1% (95% CL = 3.0, 24.3) was seen 13 days later. This association occurred in a filtered water supply in compliance with current federal standards. PMID- 9345660 TI - Socioeconomic status and health in blacks and whites: the problem of residual confounding and the resiliency of race. AB - A large number of epidemiologic studies have focused on racial/ethnic differences, particularly between blacks and whites. Because health endpoints and racial categorizations are associated with socioeconomic status, investigators generally adjust for socioeconomic indicators. The intention is usually to control for confounding, thereby making groups comparable and excluding socioeconomic status as an alternative explanation to hypotheses of innate physiologic differences. A threat to the validity of these analyses is therefore the presence of residual confounding. We identify four potential sources of residual confounding in this analytical design: categorization of socioeconomic status variables, measurement error in socioeconomic indicators, use of aggregated socioeconomic status measures, and incommensurate socioeconomic indicators. Using simulations and examples from the literature, we demonstrate that the effect of residual confounding is to bias interpretation of data toward the conclusion of independent racial/ethnic group effects. Investigators often refer to possible "genetic" differences on the basis of models that control for socioeconomic status. We propose that such conclusions on the basis of this analytical strategy are generally unwarranted. Racial/ethnic differences in disease are a pressing public health concern, but the current approach does not often provide a basis for inference about putative biological factors in the etiology of this disparity. PMID- 9345661 TI - Accounting for pregnancy dependence in epidemiologic studies of reproductive outcomes. AB - The aim of this paper is to evaluate the contribution of hierarchical mixed models to the analysis of epidemiologic studies of environmental exposure and reproductive outcomes. We have re-analyzed, with a logistic-normal mixed model, four studies investigating the relation between the frequency of spontaneous abortions and paternal or maternal environmental exposures. The data include multiple pregnancies for some women. The fitted models allow for between-woman variation of the propensity for spontaneous abortion, by including a random intercept in the logistic model to adjust for within-woman correlations on pregnancy outcomes. We have discussed and implemented two estimation methods, maximum likelihood and Bayesian inference. We found similar values in the various epidemiologic studies of the between-woman variance of the intrinsic risk of spontaneous abortion. The size of this variance corresponds to a substantial variability in risk between women. Indeed, the risk of spontaneous abortion calculated for "nonexposed" pregnancies, that is, with mother's age, birth order, tobacco consumption, and maternal environmental exposure equal to the referent class, can vary, according to this model, from 2% to 17%. PMID- 9345663 TI - Does immunosuppressive ultraviolet radiation explain the latitude gradient for multiple sclerosis? AB - Multiple sclerosis is regarded as an autoimmune disease. The autoimmune process is thought to be triggered by early-life exposure to viral/bacterial antigens that share key peptide sequences with myelin protein (the target of autoimmune attack in multiple sclerosis). It has long been known that the incidence of multiple sclerosis is positively correlated with latitude, particularly in Caucasian populations. There is no agreed explanation for this latitude gradient, however. Ultraviolet radiation level is negatively correlated with latitude. Recent evidence suggests that ultraviolet-B is immunosuppressive, affecting particularly T-cell activity and delayed-type hypersensitivity. We hypothesize here that the latitude gradient of multiple sclerosis may reflect differential ultraviolet-induced suppression of autoimmune activity, particularly since the autoimmune profile of multiple sclerosis is characterized by disturbances of those T-cell-related activities that are specifically affected by ultraviolet-B. We propose some specific tests of this hypothesis. PMID- 9345662 TI - Condom use and first-time urinary tract infection. AB - We evaluated the effects of condom use, lubricated condom use, and spermicide use on risk of acquiring first urinary tract infection in a case-control study of sexually active college women ages 18-39 years. Cases (N = 144) were women with first urinary tract infection that was confirmed by culture recruited at the student health service; controls (N = 286) were women without a history of urinary tract infection who were randomly sampled from all women enrolled at the university. Participants completed a self-administered questionnaire regarding type and frequency of condom use during the previous 2 weeks. Condoms and spermicides usually were used in combination with each other or oral contraceptives. After adjusting for frequency of vaginal intercourse, using unlubricated condoms compared with using no birth control method strongly increased the risk of first urinary tract infection (odds ratio = 29.1; 95% confidence interval = 3.1-1,335). Using a lubricated condom (with or without spermicide in the lubricant) or a spermicidal cream or gel with an unlubricated condom was associated with two- to eightfold risk of first urinary tract infection. Unlubricated condom use was strongly associated with risk of first urinary tract infection, but this effect was largely neutralized by using a spermicidal cream or gel with the unlubricated condom or by using a lubricated condom. PMID- 9345664 TI - Dioxin exposure and cancer risk: a 15-year mortality study after the "Seveso accident". AB - Dioxin (2,3,7,8-tetrachlorodibenzo-para-dioxin, or TCDD) is a powerful carcinogen in experimental animals, whereas the evidence in humans is limited. We examined cancer mortality from 1976 to 1991 among residents of Seveso, Italy, which was highly contaminated after an industrial accident. The area was divided into zones with decreasing exposure to dioxin (A = highest, B = lower, R = lowest). The population of a surrounding noncontaminated area was used as a reference group. Zone A was small (11,516 person-years); in that zone, we saw a moderate increase in mortality from digestive cancer among women [relative risk (RR) = 1.5; 95% confidence interval (CI) = 0.5-3.5]. In zone B, we also saw excesses at digestive sites (83,610 person-years), 10 years after the accident. Women had an increased mortality from stomach cancer (RR = 2.4; 95% CI = 0.8-5.7), and men had increased mortality from rectal cancer (RR = 6.2; 95% CI = 1.7-15.9). Hematologic neoplasms were increased. The highest risks were seen in zone B for leukemia in men (RR = 3.1; 95% CI = 1.3-6.4), multiple myeloma in women (RR = 6.6; 95% CI = 1.8-16.8), and Hodgkin's disease in both genders (RR = 3.3; 95% CI = 0.4-11.9 in men; and RR = 6.5; 95% CI = 0.7-23.5 in women). Soft tissue sarcoma was elevated only among zone R males (256,408 person-years; RR = 2.1; 95% CI = 0.6-5.4). We found no increase for all-cancer mortality or major specific sites (for example, respiratory among males, breast among females). The specific excesses that we observed were not explained by bias or confounding, and their association with dioxin exposure is plausible. The follow-up is continuing. PMID- 9345665 TI - Family history and risk of fatal prostate cancer. AB - To examine the relation between fatal prostate cancer and family history of prostate cancer in a first-degree relative, we analyzed data from a prospective mortality study of 481,011 men with no history of cancer at enrollment in 1982. During 9 years of follow-up, 1,922 deaths from prostate cancer occurred. Results from Cox proportional hazard models showed that family history of prostate cancer was related to fatal prostate cancer [rate ratio (RR) = 1.60; 95% confidence interval (CI) = 1.31-1.97]; men with two or more affected relatives had a greater than threefold increase in risk (RR = 3.19; 95% CI = 1.51-6.71). Men whose relatives were diagnosed with prostate cancer before age 65 years (RR = 2.03; 95% CI = 1.33-3.09) had a greater effect of family history than men whose relatives were diagnosed at older ages (RR = 1.50; 95% CI = 1.17-1.91). Rate ratios did not increase with decreasing age of the study participants. The 60% increase in risk for men with at least one affected relative is lower than that reported in previous studies. PMID- 9345666 TI - Dietary fiber and colorectal cancer risk. AB - We conducted a population-based case-control study among different ethnic groups in Hawaii to evaluate the role of various types and components of fiber, as well as micronutrients and foods of plant origin, on the risk of colorectal cancer. We administered personal interviews to 698 male and 494 female Japanese, Caucasian, Filipino, Hawaiian, and Chinese cases diagnosed during 1987-1991 with adenocarcinoma of the colon or rectum and to 1,192 population controls matched to cases by age, sex, and ethnicity. We used conditional logistic regression to estimate odds ratios, adjusted for caloric intake and other covariates. We found a strong, dose-dependent, inverse association in both sexes with fiber intake measured as crude fiber, dietary fiber, or nonstarch polysaccharides. We found inverse associations of similar magnitude for the soluble and insoluble fiber fractions and for cellulose and noncellulosic polysaccharides. This protective effect of fiber was limited to fiber from vegetable sources, with an odds ratio of 0.6 (95% confidence interval = 0.4-0.9) and 0.5 (95% confidence interval = 0.3 0.7) for the highest compared with the lowest quartile of intake for men and women, respectively. We found associations of the same magnitude for soluble and insoluble vegetable fiber, but no clear association with fiber from fruits or cereals. This pattern was consistent between sexes, across segments of the large bowel (right colon, left colon, and rectum), and among most ethnic groups. The effect of vegetable fiber may be independent of the effects of other phytochemicals, since the effect estimates remained unchanged after further adjustment for other nutrients. Intakes of carotenoids, light green vegetables, yellow-orange vegetables, broccoli, corn, carrots, bananas, garlic, and legumes (including soy products) were inversely associated with risk, even after adjustment for vegetable fiber. The data support a protective role of fiber from vegetables against colorectal cancer, which appears independent of its water solubility property and of the effects of other phytochemicals. The data also indicate that certain vegetables and fruits may be protective against this disease through mechanisms other than their fiber content. PMID- 9345667 TI - The waiting time distribution as a graphical approach to epidemiologic measures of drug utilization. AB - The emergence of large, computerized pharmacoepidemiologic databases has enabled us to study drug utilization with the individual user as the statistical unit. A recurrent problem in such analyses, however, is the overwhelming volume and complexity of data. We here describe a graphical approach that effectively conveys some essential utilization parameters for a drug. The waiting time distribution for a group of drug users is a charting of their first prescription presentations within a specified time window. For a drug used for chronic treatment, most current users will be captured at the beginning of the window. After a few months, the graph will be dominated by new, incident users. As examples, we present waiting time distributions for insulin, ulcer drugs, systemic corticosteroids, antidepressants, and disulfiram. Appropriately analyzed and interpreted, the waiting time distributions can provide information about the period prevalence, point prevalence, incidence, duration of use, seasonality, and rate of prescription renewal or relapse for specific drugs. Each of these parameters has a visual correlate. The waiting time distributions may be an informative supplement to conventional drug utilization statistics, and possibly also a useful screening tool for unusual prescribing patterns. PMID- 9345668 TI - Mortality in current and former users of clozapine. AB - Clozapine (Clozaril), a tricyclic dibenzodiazepine, causes fewer extrapyramidal side effects than do other antipsychotic drugs. Because it can induce agranulocytosis, however, clozapine is indicated only for schizophrenia that is not responsive to other therapies. To describe the drug's effects on mortality, we compared rates of various causes of death in 67,072 current and former clozapine users. We linked data from a national registry of clozapine recipients to the National Death Index and Social Security Administration Death Master Files, obtained death certificates, and calculated mortality rates for underlying causes of death using standardization to adjust for age, sex, and race. During 1991-1993, there were 396 deaths in 85,399 person-years for patients ages 10-54 years. Mortality was lower during current clozapine use than during periods of non-use. Mortality from suicide was decreased in current clozapine users by comparison with past users [rate ratio (RR) = 0.17; 95% confidence interval (CI) = 0.10-0.30]. During clozapine use, there were elevations in mortality rates for less common causes of death, including pulmonary embolism (RR for current exposure compared with past clozapine use = 5.2) and respiratory disorders (RR = 2.9). Clozapine appears to reduce mortality in severe schizophrenics, mostly by decreasing suicide rates. PMID- 9345670 TI - Determinants of risk of spontaneous abortions in the first trimester of pregnancy. AB - Several factors, such as socioeconomic status, obstetrical and menstrual history, and contraceptive methods, have been associated with risk of spontaneous abortion. We conducted a hospital case-control study to analyze risk factors for spontaneous abortion during the first trimester. Cases were 782 women admitted for spontaneous abortion. Controls included 1,543 women who gave birth at term in the same hospitals. Adjusted odds ratios (ORs) for spontaneous abortion were 0.9 and 0.6, respectively, for women reporting 7-11 and > or = 12 years of schooling, compared with women reporting < 7 years of education. A history of pelvic inflammatory disease increased the odds ratio fivefold [OR = 5.1; 95% confidence interval (CI) = 1.0-26.2]. The OR for spontaneous abortion was 1.7 (95% CI = 1.4 2.1) in women reporting previous spontaneous abortions. PMID- 9345669 TI - Is infant immunization a risk factor for childhood asthma or allergy? AB - The Christchurch Health and Development Study comprises 1,265 children born in 1977. The 23 children who received no diphtheria/pertussis/tetanus (DPT) and polio immunizations had no recorded asthma episodes or consultations for asthma or other allergic illness before age 10 years; in the immunized children, 23.1% had asthma episodes, 22.5% asthma consultations, and 30.0% consultations for other allergic illness. Similar differences were observed at ages 5 and 16 years. These findings do not appear to be due to differential use of health services (although this possibility cannot be excluded) or con-founding by ethnicity, socioeconomic status, parental atopy, or parental smoking. PMID- 9345671 TI - From the Shy trial defense: a leak, or red herring? PMID- 9345672 TI - Conflict of interest. PMID- 9345673 TI - Alcohol, coronary heart disease, and mortality. PMID- 9345674 TI - Polychlorinated biphenyls, chlordanes, and the etiology of non-Hodgkin's lymphoma. PMID- 9345675 TI - Epidemiology and the Internet. PMID- 9345676 TI - Identifying deaths using the Social Security Administration Death Master Files. PMID- 9345677 TI - Identifying deaths using the Social Security Administration Death Master Files. PMID- 9345678 TI - Chiropractic in Sweden: a short description of patients and treatment. AB - BACKGROUND: Most information on chiropractors and chiropractic emanates from North America. With an increasing number of chiropractors in Europe, it is important to produce relevant documentation concerning the European practice of chiropractic. OBJECTIVES: To describe the typical Swedish chiropractic patient and the treatment he/she receives. DESIGN: Chiropractors interviewed 10 consecutive patients using a standardized questionnaire. OUTCOME VARIABLES: Age, sex, previous treatment by chiropractor, area and duration of complaint, area and type of treatment and number of return visits (truncated data). RESULTS: Of the 86 chiropractors who could take part in the study, 78% participated. They each collected information on 10 consecutive patients (n = 628, response rate 73% of target sample), altogether 1858 return visits. Most patients were aged between 25 and 64 yr, with no difference in numbers between genders. Typically, they sought care for low back pain of up to 1 month's duration, were treated with spinal manipulation and received 2-3 treatments. Swedish chiropractic patients thus seem to seek care relatively early and undergo a short-lasting treatment program. However, a larger number of treatments was given to patients with problems of longer duration; patients who had not previously consulted a chiropractor presented with more long-lasting problems. CONCLUSIONS: The findings are in line with present-day concepts that emphasize the concurrent needs to avoid both the development of chronicity and long-lasting, invasive clinical intervention. Chiropractic care seems not to be a first choice therapy for those who have not previously received chiropractic care. PMID- 9345680 TI - Interexaminer reliability in physical examination of the neck. AB - BACKGROUND: There are numerous clinical tests used in the evaluation of patients with symptoms arising from the cervical spine. It is necessary to use clinical tests with high validity and reliability. Previous studies of reliability of clinical tests used in the evaluation of the cervical spine have come to various conclusions, most of which suggest low reliability. This might be explained by differences between examiners in performance and where the limit of normality is placed. OBJECTIVE: To evaluate the interexaminer reliability of clinical tests used in everyday clinical work, where the examiners base their evaluations on a comparison between left and right sides. STUDY DESIGN: A total of 50 volunteers were examined by two physiotherapists. The interexaminer reliability of clinical tests included in the physical examination of patients with symptoms from the cervical spine was evaluated. METHODS: Two physiotherapists independently examined volunteers. RESULTS: An acceptable reliability was found for two of 10 clinical tests. CONCLUSION: When it is possible to compare left and right sides, it is possible to show acceptable reliability for some clinical tests. Reliability studies most often find low reliability, perhaps because of bias; clinical tests are not standardized. In future studies, greater efforts should be taken to reduce bias. PMID- 9345679 TI - Side effects of chiropractic treatment: a prospective study. AB - OBJECTIVES: To investigate whether the characteristics of unpleasant sid effects after spinal manipulative therapy coincide with those obtained in a previous study. DESIGN: A prospective interview survey using standard questionnaires. SETTING: Sixty-six Swedish private practices of chiropractic (response rate, 78%). SUBJECTS: Ten consecutive patients per chiropractor (625 patients, 73% of target sample; 1858 recorded visits). INTERVENTION: Spinal manipulation. MAIN OUTCOME MEASURES: Self-reported unpleasant reactions, time of onset, duration and severity of symptoms. RESULTS: Reactions to spinal manipulation are common and benign. They typically arise and disappear shortly after treatment (usually gone the day after treatment). The most common reactions are local discomfort in the area of treatment (two thirds of reactions), followed by pain in areas other than that of treatment, fatigue or headache (10% each). Nausea, dizziness or "other" reactions are uncommonly reported (< 5% of reactions). Reactions are most commonly reported by women and (for both genders) at the beginning of the treatment series. Patients with long lasting problems are more likely to report treatment reactions; however, patients with no prior experience of chiropractic care do not report more reactions than patients previously treated by chiropractors. CONCLUSION: Common and uncommon reactions to chiropractic spinal manipulation have been identified, are to a large degree foreseeable and seem to be predominantly physiological in nature. PMID- 9345681 TI - Application and evaluation of the kappa statistic in the design and interpretation of chiropractic clinical research. AB - The Kappa statistic (K) is becoming more widely used in chiropractic clinical research. Although the calculation of K is straightforward, assumptions made regarding the acquisition and interpretation of data require serious consideration before conclusions regarding the final outcome can be made. Of particular interest is the application of K for the interpretation of validity and reliability. In this regard, I argue against the use of K as a summary statistic. I present here a description of K as a function of observer agreement (P(o)), without which K has little meaning, and show how a knowledge of this function should direct the design and, therefore, the interpretation of chiropractic clinical research. Only when all concerns regarding experimental design are either addressed directly or compensated for in the analysis can interpretations be made regarding validity or reliability. Lacking such attention, K becomes an indicator of experimental design rather than an indicator of validity or reliability. PMID- 9345683 TI - Clinical consequences of spina bifida occulta. AB - OBJECTIVE: To study data concerning the pathogenesis, frequency of occurrence, clinical manifestations and associated abnormalities of spina bifida occulta (SBO) and re-evaluate the clinical importance of the lesion. DATA SOURCES: International journal articles indexed through Medline, and specific related texts, the majority of which were published after 1989. Key indexing terms used were spina bifida occulta, tethered cord syndrome and spondylolysis. RESULTS: The reported frequency of occurrence of SBO varies widely, depending largely on the age groups included in a particular study. The most accurate estimate of occurrence rate is 17% of examined spines. There is a significant association of some cutaneous stigmata, most notably hypertrichosis, with midline posterior arch defects. An increasing amount of evidence links SBO with a number of specific anomalies and clinical syndromes, including intraspinal lipoma, tethered cord syndrome, genitourinary dysfunction, increased incidence of disc pathology, lumbar spondylolysis, foot deformities and syringomyelia. A questionable association exists with epilepsy. A supposed link between constipation and SBO is lacking sufficient data to support it. CONCLUSIONS: SBO may be associated with pathology and significant sequelae, although the majority of lesions pose no clinical threat. The predictive value for adverse sequelae in a particular lesion is difficult to assess; however, multilevel occurrence and more expansive involvement in a given segment seem to be associated with higher risk of sequela. The treatment for SBO with progressive neurologic deficit is surgical intervention; however, reversal of the deficit is unusual and a halting of neurologic deterioration is a more realistic goal. Early diagnosis of this lesion, before the age of 3 yr, is associated with better surgical outcomes. PMID- 9345684 TI - Protruded lumbar intervertebral nucleus pulposus in a 12-year-old girl who recovered after nonsurgical treatment: a follow-up case report. AB - OBJECTIVE: To discuss the nonsurgical treatment of a lumbar intervertebral disc protrusion with obvious neurological irritation in a juvenile patient. CLINICAL FEATURES: A 12-yr-old girl suffered from low back and right leg pain that came on after a lengthy walk. There was a limp on the right, with pain also radiating to the right leg. A diagnosis of lumbar disc herniation was made after advanced imaging methods were applied. INTERVENTION AND OUTCOME: The girl underwent special spinal rotational manipulation, epidural injection and, later on, physical exercises. She did not respond to the manipulation as adult patients with similar signs do; in her case, response did not occur until nearly 1 month of hospitalization. A follow-up study with computed tomography, plain and dynamic radiology and thermography found that the protruded nucleus pulposus was not changed in size and position and the functional recovery of her lumbar spine was far from fulfilled until 8 months after being dismissed from the hospital. CONCLUSION: Juvenile patients suffering from lumbar disc protrusion are rare and seldom undergo conservative treatments, especially when obvious neurological irritation occurs. This case shows the difficulties in managing the juvenile patient with conservative care. Doctors or specialists should always be cautious about manipulating juvenile patients with too much force. PMID- 9345682 TI - Changes in brain function after manipulation of the cervical spine. AB - OBJECTIVE: To ascertain whether manipulation of the cervical spine is associated with changes in brain function. DESIGN: Physiological cortical maps were used as an integer of brain activity before and after manipulation of the cervical spine in a large (500 subjects), double-blind controlled study. SETTING: Institutional clinic Participants: Adult volunteers. INTERVENTION: Five hundred subjects were divided into six comparative groups and underwent specific manipulation of the second cervical motion segment. Blinded examiners obtained reproducible pre- and postmanipulative cortical maps, which were subjected to statistical analysis. MAIN OUTCOME MEASURES: Brain activity was demonstrated by reproducible circumferential measurements of cortical hemispheric blind-spot maps before and after manipulation of the second cervical motion segment. Twelve null hypotheses were developed. The critical alpha level was adjusted in accordance with Bonferroni's theorem to .004 (.05 divided by 12) to reduce the likelihood of wrongly rejecting the null hypothesis (i.e., committing a Type I error). RESULTS: Manipulation of the cervical spine on the side of an enlarged cortical map is associated with increased contralateral cortical activity with strong statistical significance (p < .001). Manipulation of the cervical spine on the side opposite an enlarged cortical map is associated with decreased cortical activity with strong statistical significance (p < .001). Manipulation of the cervical spine was specific for changes in only one cortical hemisphere with strong statistical significance (p < .001). CONCLUSIONS: Accurate reproducible maps of cortical responses can be used to measure the neurological consequences of spinal joint manipulation. Cervical manipulation activates specific neurological pathways. Manipulation of the cervical spine may be associated with an increase or a decrease in brain function depending upon the side of the manipulation and the cortical hemisphericity of a patient. PMID- 9345685 TI - Abdominal aortic aneurysms: clinical diagnosis and management. AB - OBJECTIVE: To review the presentation, diagnosis and management of aortic aneurysms. Case reports and a brief topic review are presented. CLINICAL FEATURES: Three cases of aneurysm that were diagnosed in a chiropractic office are discussed. An aneurysm is defined as an abnormal dilation of the aorta as a result of atherosclerosis, genetic predisposition and/or acquired biochemical alterations in the wall of the aorta. The "classic triad," hypotension, back pain and a pulsatile abdominal mass are present in only 50% of those people with ruptured abdominal aortic aneurysms (AAA). Large unruptured aneurysms are quite often asymptomatic and are often found incidentally on physical or X-ray examination. History, palpation, auscultation and imaging are all helpful in diagnosing AAAs, and all are readily available in a chiropractic office. INTERVENTION AND OUTCOME: Surgical intervention is generally considered appropriate in AAAs > 5 cm in diameter. All patients recovered after surgical repair of the aneurysm. CONCLUSION: Chiropractors can perform simple diagnostic procedures to differentially diagnosis and screen for AAAs. Such screening measures may yield a statistical decrease in deaths caused by rupture of aortic aneurysms. PMID- 9345686 TI - The dynamics of composite licensing boards from the chiropractic perspective. PMID- 9345687 TI - The cervical subluxation and regional cerebral blood flow. PMID- 9345688 TI - Safety in chiropractic practice. Part II: Treatment to the upper neck and the rate of cerebrovascular incidents. PMID- 9345689 TI - Safety in chiropractic practice. Part II: Treatment to the upper neck and the rate of cerebrovascular incidents. PMID- 9345690 TI - Asymmetrical hemispheric control of visual-spatial attention in adults with attention deficit hyperactivity disorder. AB - As neuropsychological mechanisms for attention have been hypothesized to be located in the right hemisphere of the brain, several investigators have begun to conceptualize attention deficit hyperactivity disorder (ADHD)-related attentional deficits as involving right-hemispheric abnormalities. The authors evaluated and compared adult patients diagnosed with ADHD with a non-ADHD group of patients using a chronometric visual-spatial attention task that is sensitive to hemispheric differences in efficiency of information processing. Reaction times across different cuing conditions, cue-target delays, and visual fields were assessed. When participants' attention was misdirected with cues in the right visual field and attention had to be switched to a target on the left visual field, there was a longer delay among ADHD adults than non-ADHD adults, specifically when the interval between the cue and target was 800 ms as compared with 100 ms. This specific pattern of dysfunction was interpreted as a difficulty with maintaining attention possibly associated with anterior attention mechanisms in the right hemisphere. PMID- 9345691 TI - Concurrent eyeblink classical conditioning and rotary pursuit performance: implications for independent nondeclarative memory systems. AB - The authors examined whether 2 nondeclarative tasks, simple eyeblink classical conditioning (EBCC) and rotary pursuit (RP), would interfere with each other when performed simultaneously. In Experiment 1,100 participants were assigned to 1 of 5 groups: paired EBCC/RP, unpaired EBCC/RP, paired EBCC as a single task, unpaired EBCC as a single task, and RP as a single task. Participants in the paired EBCC/RP group showed significantly greater acquisition of conditioned responses than did participants in the unpaired EBCC/RP group, and the unconditioned eyeblink response was similar in both groups. Comparisons of the paired EBCC/RP and paired EBCC-as-a-single-task groups indicated no differences in trials to criterion, but on some measures the single-task group conditioned better. Controls introduced in Experiment 2 did not change this pattern. Results provide some evidence for the lack of interference between EBCC and RP. PMID- 9345693 TI - Syntactic and semantic processing in schizophrenic patients evaluated by lexical decision tasks. AB - Two lexical-decision tasks with 500-ms stimulus-onset asynchrony were conducted with 34 schizophrenic patients. This group consisted of 24 schizophrenic patients with thought disorder (TD) and 10 schizophrenic patients without thought disorder (NTD), 14 psychiatric controls (depressive illness), 20 hospitalized controls, and 20 normal controls. One lexical-decision task with semantic relations (related vs. unrelated, Experiment 1) and 1 task with syntactic relations (congruent vs. incongruent; Experiment 2) were used to evaluate processing of different lexical information. In Experiment 1, although all control groups and NTD schizophrenic patients showed semantic priming, TD schizophrenic patients did not. In Experiment 2, all groups showed a significant syntactic effect. These findings provide evidence for an abnormality in semantic processing and the preservation of syntactic processing in TD schizophrenic patients, thus suggesting a deficit in the processing of semantic information under certain conditions when compared with normal syntactic processing. PMID- 9345692 TI - Comparing the difficulty of letter, semantic, and name fluency tasks for normal elderly and patients with Parkinson's disease. AB - Research on the effect of Parkinson's disease (PD) on verbal fluency has produced conflicting results. In this study, 88 PD patients with no dementia, 11 PD patients with questionable mental status, 15 PD patients with dementia, and 46 elders free from mental disorder were administered a variety of semantic, letter, and name fluency tasks. The results revealed that, contrary to popular assumption, semantic fluency was not always superior to letter fluency. Rather, verbal fluency was influenced by the nature of the individual categories. Interestingly, the relative difficulty of many categories was fairly stable across groups. The results also indicated that the individual fluency tasks were differentially sensitive to the mental status of the PD patients. Overall, the findings suggest that closer attention to the nature of the tested categories may help clarify the inconsistent effects of PD on verbal fluency. PMID- 9345694 TI - Automatic versus controlled semantic priming in schizophrenia. AB - Schizophrenic individuals (n = 31), including paranoid and nonparanoid diagnostic subgroups, and normal controls (n = 20) participated in a semantic priming experiment involving a single-choice lexical decision task. For the automatic priming blocks, a 260-ms stimulus onset asynchrony (SOA) was used; for the controlled priming blocks, 1,000-ms SOA was used. The paranoid subgroup showed significantly less priming than did the control group. The nonparanoid subgroup showed a decrease in priming compared with the control group that approached significance. There was an increased priming effect for the controlled compared with the automatic priming condition; this difference was not modulated by participant group. Nonsignificant semantic priming (equal to 0) occurred only for schizophrenic subgroups and only in automatic priming conditions. PMID- 9345695 TI - Fluency and memory differences between ischemic vascular dementia and Alzheimer's disease. AB - This study compared 32 patients with ischemic vascular dementia (IVD) to 32 patients with probable Alzheimer's disease (AD) on select language and verbal memory tests. The IVD and AD patients were individually matched on the basis of age, dementia severity, years of education, and gender. The IVD patients had poorer verbal fluency, but better free recall, fewer recal intrusions, and better recognition memory than the AD patients. Relationships between the neuropsychological measures and radiological indexes of cortical and subcortical pathology were also examined. Number of infarcts, white-matter lucency, and ventricular enlargement correlated with some of the neuropsychological measures; cortical atrophy correlated with most of the measures. The findings suggest that neuropsychological deficits in IVD may be related to dysfunction of frontal subcortical circuits, although an associated degenerative cortical process may also be involved. PMID- 9345697 TI - Executive functions in multiple sclerosis: an analysis of temporal ordering, semantic encoding, and planning abilities. AB - Previous studies have consistently demonstrated impairments in conceptual reasoning and set-shifting abilities in patients with multiple sclerosis (MS). Other executive functions have been less frequently examined. We compared 44 MS patients and 48 demographically matched controls on a temporal-ordering and semantic-encoding task and on a test of planning (Tower of Hanoi). Compared with controls, MS patients experienced deficient semantic encoding and planning but unimpaired temporal-order memory. For both tasks, post hoc analyses indicated that chronic-progressive MS patients contributed most to the group differences. A combination of poor planning and slowed information-processing speed was hypothesized to have contributed to MS patients' impaired Tower of Hanoi performance. Further research is needed to explore the possible relationship between semantic-encoding and planning deficits in MS and social and occupational disabilities. PMID- 9345698 TI - Indirect influence of modality on direct memory for words and their modality: closed-head-injured and control participants. AB - Twenty closed-head-injured (CHI) patients and 28 control participants were tested on recall and recognition of words. In addition, memory for modality (i.e., visual vs. auditory) of word presentation was measured directly (i.e., recognition) and indirectly (i.e, by its influence on word and modality recognition). As predicted, the CHI patients were impaired relative to controls on all of the direct memory tasks; that is, word recall, word recognition, and modality judgment. However, the CHI and control groups did not differ significantly on the magnitude of the modality effect (i.e., facilitation due to correspondence of modality in learning and test). The findings are interpreted in the theoretical framework that distinguishes between item (i.e., words) and source (i.e., modality) memory and between direct and indirect measures of memory. PMID- 9345696 TI - Perseverative behavior in Alzheimer's disease and subcortical ischemic vascular dementia. AB - Perseverative behavior has not been extensively studied in patients with dementia. In this study, perseverative behavior was elicited with the dementia version of the Graphical Sequence Test. A control group and participants with Alzheimer's disease (AD) and subcortical ischemic vascular dementia (IVD) were studied. A factor analysis revealed a 3-factor model consisting of perseverations related to semantic knowledge, motor functioning, and a third, intermediary factor. IVD participants made more total perseverations than did AD participants. Perseverations made by AD participants were correlated with deficits on tests of semantic knowledge, whereas the perseverations made by IVD participants were correlated with motor and frontal systems tests. Results are consistent with the view that perseverative behavior is hierarchically arranged in terms of specific levels of cognitive complexity and the overall pattern of cognitive deficits associated with each type of dementia. PMID- 9345699 TI - Developmental instability and cerebral lateralization. AB - On the basis of prior studies of handedness, it was predicted that variations from modal asymmetry scores on cognitive tasks, in either direction from the mean, would be associated with an elevated incidence of classic markers of developmental instability (minor physical anomalies and fluctuating anatomic asymmetries). University students (N = 146) were administered 4 tasks that typically reveal functional asymmetries: the fused rhymed words dichotic listening task, the line bisection task, the chimeric faces task, and the cartoon faces task. A composite measure of developmental instability was computed from minor physical anomalies and fluctuating asymmetries. Participants with greater evidence of developmental instability had more atypical lateralization scores, deviating more from the sample mean, in either direction. Directional asymmetries were unrelated to developmental instability. These results suggest that developmental instability influences variation in the lateralization of cognitive skills as well as handedness. PMID- 9345700 TI - Individual differences in lateralization: effects of gender and handedness. AB - Male and female left- and right-handers participated in 3 experiments designed to investigate 3 components of performance asymmetry in lateralized tasks. Experiment 1 used a consonant-vowel-consonant (CVC) identification task measuring quantitative differences in hemispheric abilities and hemispheric control and qualitative differences in hemispheric strategies. The quantitative data revealed that left-handers have a smaller performance asymmetry than do right-handers and that both groups have the same degree of increased accuracy when stimuli are presented bilaterally. Handedness affected the qualitative measures of men, not of women. Experiment 2 used nominal and physical letter-matching tasks with bilateral presentations and measured the flexibility of callosal function. The results suggest that left-handers have less flexible interhemispheric communication than do right-handers and show no effect of gender. Experiment 3 used a chair identification task indexing hemispheric arousal bias. Left-handers tended to have more aroused right than left hemispheres, whereas the distribution of right-handers was centered around 0 arousal bias. Intertask analyses revealed a relationship between arousal bias and metacontrol, where individuals with more aroused right hemispheres tended to use a right-hemisphere strategy in the bilateral condition of the CVC experiment. Intercorrelations between measures from the experiments revealed only a limited relationship between metacontrol patterns in the CVC task and a measure of callosal flexibility in the physical letter-matching task. The results are discussed in the context of the relationships between dimensions of hemispheric asymmetry. PMID- 9345701 TI - Developmental sex differences in verbal learning. AB - Although sex differences in verbal learning and memory have been reported in adults, much less is known about when these sex differences emerge and how they develop. In this study, 401 boys and 410 girls between the ages of 5 and 16 years were administered the California Verbal Learning Test--Children's Version. Sex differences were found at all age levels. Girls performed better than boys on all of the immediate and delayed recall trials and on the delayed recognition trial. Girls were also more likely than boys to use a semantic clustering strategy and displayed more effective long-term memory mechanisms. Boys made more intrusion errors and displayed greater vulnerability to interference between the 2 test lists. Because boys had higher mean scores on Wechsler Intelligence Scale for Children--Revised Vocabulary, the observed female superiority in verbal learning could not be attributed to sex differences in overall word knowledge. PMID- 9345702 TI - Role of the hippocampus in sex differences in verbal memory: memory outcome following left anterior temporal lobectomy. AB - The authors examined the neural and cognitive bases for sex differences in verbal memory in 57 patients who underwent left anterior temporal lobectomy (ATL) for the treatment of intractable seizures. On the California Verbal Learning Test (D. C. Delis, J. H. Kramer, E. Kaplan, & B. A. Ober, 1987), women recalled more words than men both before and after surgery, regardless of the extent of hippocampal damage. Extent of hippocampal sclerosis was related to memory loss in both men and women. Women's superiority in verbal memory appears to result in part from their use of an efficient encoding strategy. Women were more likely than men to use semantic clustering both before and after ATL, and sex differences in word recall were attenuated after scores were adjusted for semantic clustering. There was no effect of ATL on semantic clustering. Taken together, these results suggest that sex differences in verbal memory are not due to differences in the integrity of the left hippocampus. PMID- 9345703 TI - Neuropsychological abnormalities among HIV-infected individuals in a community based sample. AB - The purpose of this study was to determine the nature and extent of neuropsychological abnormalities among HIV-infected individuals and to examine the interrelationships between measures of cognitive functions and the factors that predict neuropsychological abnormalities. The study focused on cross sectional data gathered in a multidisciplinary research clinic form 200 HIV infected (HIV +) men and women recruited from primary medical care settings. Composite scores representing six cognitive domains were derived from the neuropsychological test data. Scores of memory, fluency, spatial, and frontal functions could be predicted by independent assessment of participants' verbal and psychomotor speed abilities. Basic verbal ability itself was predicted by education, race, and handedness, whereas speed was predicted by age, CD4+ cell counts, and a lifetime history of major depression. This model of effects is consistent with the hypothesis that psychomotor slowing is central to mild cognitive disorder in HIV infection and that such changes are associated with markers of the severity of systemic infection. PMID- 9345704 TI - Reliability, performance characteristics, construct validity, and an initial clinical application of a visual object learning test (VOLT). AB - Whereas verbal learning has received considerable attention by cognitive neuropsychology, spatial object learning has been more resistant to study. The paucity of visual learning data has hampered attempts to clarify if visual learning has unique features with specialized neural substrates. In schizophrenia, severe verbal learning impairment has been established, but lack of comparable visual learning measures has thwarted the dissociation of verbal and visual abilities. The Visual Object Learning Test (VOLT) was developed to examine aspects of visual-spatial learning and memory in a manner analogous to available verbal tests. Studies were performed to establish normative performance characteristics, convergent and divergent validity, and the sensitivity of the VOLT to detection of individual differences in normal (through sex and age) and pathologic variability (through persons with schizophrenia). The results indicated excellent internal consistency, convergent and divergent validity, and sensitivity to the effects of aging and pathology. Persons with schizophrenia were impaired in both learning and retention. The authors conclude that memory impairment in schizophrenia may not be specific to verbal learning. PMID- 9345705 TI - Test-retest stability of the California verbal learning test in older persons. AB - One hundred fifty-one healthy older persons were administered the California Verbal Learning Test (CVLT) on 2 occasions, an average of 1.30 years apart. Means for age and education were 69.63 years (SD = 6.46) and 14.91 years (SD = 2.54), respectively. Stability coefficients ranged from .24 on number of perseverations to .76 for both List A total recall and long-delay free recall. Only 5 CVLT scores demonstrated a significant change on retest, and the overall magnitude of improvement was small. To assist clinicians in the process of detecting significant change on retest, the authors provide standard error of difference values and 90% confidence intervals for 22 CVLT scores. PMID- 9345706 TI - The impact of thrombopoietin and related Mpl-ligands on transfusion medicine. PMID- 9345707 TI - The computer or electronic crossmatch. PMID- 9345708 TI - Umbilical cord blood as a source of progenitor cells to reconstitute hematopoiesis. PMID- 9345709 TI - Consent for transfusion: is it informed? AB - Informed consent for transfusions presumes that the patient has been "informed," meaning they have been given sufficient information to make an intelligent choice, and that they "consent," meaning that the patient is competent and free to consent. Although this may be oral or written, the latter is preferable for documentation and legal proof. This may either be as a chart note, or form. In any case, the informed transfusion recipient should be given a description of the procedure in lay terms, told of the expected benefit, including the outcome without the transfusion, and what reasonably foreseeable adverse consequences may occur. The individual should have the opportunity to ask questions of the physician performing the informed consent process. There should be time to provide the information and a discussion; and the patient must be able to comprehend all of this. If this process is performed by the physician talking to his/her patient, then, the patient is likely to be informed about, and consent to, elective transfusions. Once obtained and documented, that consent should be presumed to be valid with a course of therapy for transfusions. If some event significantly changes some aspects of the information on which the patient relied in consenting and now should be aware of, then, the patient should again give consent for transfusion, after being informed of this new information. PMID- 9345710 TI - Liquid cold storage of platelets: a revitalized possible alternative for limiting bacterial contamination of platelet products. PMID- 9345711 TI - The alloimmune thrombocytopenic syndromes. AB - We have summarized five thrombocytopenic syndromes caused by platelet-reactive alloantibodies. Increased awareness of these five syndromes, together with the greater availability of highly-specialized laboratory methods to detect and to characterize platelet-reactive alloantibodies, will lead to their more frequent diagnosis. It is important for the clinician to consider unusual alloimmune thrombocytopenic disorders in clinical settings in which alloantigens that could be limited to just one family could cause important disease (ie, NAT caused by a private alloantigen; theoretically, PAT or TAT related to directed donations of blood or bone marrow). These considerations underscore the need for serological investigations to involve the family members rather than to rely on standard platelet typing donor pools. PMID- 9345712 TI - Changes in some red blood cell and clinical laboratory parameters in young and old Beagle dogs. AB - Routine laboratory and lipid peroxidase parameters in the blood of Beagle dogs under 1 year of age (7 males, 7 females) and over 9 years of age (7 males, 7 females) were compared. The mean corpuscular haemoglobin (MCH) and haemoglobin (Hb) values, plasma total protein (TP) and globulin concentrations in the older dogs were significantly higher than in the younger ones by 13%, 6%, 10% and 15%, respectively. The plasma inorganic phosphate concentration, however, was 30% lower in the older dogs than in the young ones. The concentration of malondialdehyde (MDA) was 30% higher, while the glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities were around twice as high in the older dogs as in the young ones. However, in the older males the concentration of reduced glutathione (GSH) was 43% and in the older females 9% less than in the young ones. Extremely high levels of MDA and low levels of GSH were only found in the older males. GSH-Px and SOD activities were significantly higher in the older male dogs than in the young ones. The activities of these enzymes were highest in the older females. From our results, we suspect that old dogs, especially males, are particularly exposed to the harmful effects of free radicals and lipid peroxidation. PMID- 9345713 TI - Serum fructosamine: a reference interval for a heterogeneous canine population. AB - Eighty-nine healthy dogs (44 males, 45 females) of different breeds, 1-12 years of age, living under varied feeding and environmental conditions, were sampled to evaluate a reference interval for serum fructosamine using a nitroblue tetrazolium photocolorimetric method. The analytical assay was evaluated by calculation of within-run and between-day variations. The results were approximately normally distributed and the calculated reference interval was 192.6-357.4 mumol/L (mean 275.0 mumol/ L, standard deviation 41.2 mumol/L). No significant differences attributable to sex or age were observed. This reference interval is wider than those previously reported in less heterogeneous groups of dogs and in those from other geographical zones. The fructosamine values in serum from 3 diabetic dogs all exceeded the upper limit of the reference interval. PMID- 9345714 TI - Failure of lipopolysaccharides to directly trigger the chemiluminescence response of isolated equine polymorphonuclear leukocytes. AB - Divergent results have been reported on the effects of lipopolysaccharides (LPS) on the activation of equine polymorphonuclear leukocytes (PMN). We therefore attempted to determine whether LPS alone can stimulate equine PMN or whether plasma factors are necessary. PMN were isolated from citrated blood on a discontinuous density gradient of Percoll. The luminol (10(-3) mol/L)-enhanced chemiluminescence (CL) of 1.25 x 10(6) cells was measured after addition of Escherichia coli LPS (0.001-10 micrograms/ml) alone or after incubation in autologous plasma (1 h, 37 degrees C). After direct stimulation with LPS, there were random variations of CL in 16 horses that were not reproducible from one sample to the next for the same horse. LPS which had been incubated in plasma gave a dose-dependent stimulation of the CL of the PMN, which did not occur if the plasma had been heat inactivated (1 h, 56 degrees C). These results indicated a role for plasma factors, which were unlikely to be cytokines, as there were no monocytes or lymphocytes in the plasma incubated with LPS, but might have been complement fragments or LPS ligands, such as LPS binding protein. Studies using specific antibodies against these factors are needed to clarify this question. PMID- 9345715 TI - Multiple anthelmintic resistance in Haemonchus contortus on a sheep farm in Kenya. AB - Multiple resistance to albendazole, thiophanate, levamisole and orally administered invermectin was detected in an isolate of Haemonchus contortus in sheep on a farm where benzimidazole resistance had already been identified. Following a faecal egg count reduction test, this was confirmed by both critical and controlled anthelmintic tests. Different groups of sheep infected naturally or given an experimental infection with the benzimidazole-resistant isolate were treated with the recommended doses of various anthelmintics. Compared to the control group, the percentage reductions in the faecal egg counts of sheep treated with albendazole, thiophanate, levamisole and ivermectin varied between 38.2% and 79.1% and the residual worm counts between 27.3% and 57.5%. The results indicate the presence of multiple anthelmintic resistance in this isolate of H. contortus. Sheep treated with closantel showed 100% reductions in faecal egg and worm counts, indicating that this drug was very effective against the population of H. contortus on the farm. PMID- 9345716 TI - The efficacy of closantel and rafoxanide against fenbendazole- and levamisole resistant Haemonchus contortus in small ruminants. PMID- 9345718 TI - Light microscopy of the enteric nervous system of horses with or without equine dysautonomia (grass sickness): its correlation with the motor effects of physostigmine. AB - Light microscopy was undertaken on sections from the caudal flexure of the duodenum and the terminal ileum proximal to the ileocaecal fold in 5 control horses, 5 horses with acute grass sickness (AGS), and 5 horses with chronic grass sickness (CGS). With the exception of the ileal submucous plexus of the CGS group, the AGS group had the lowest number of neurons as measured using a subjective scoring scheme. The proportion of abnormal neurons in the AGS group was similar in both plexuses and both regions, whereas the values for the CGS group were much higher in the duodenal region than in the ileal region. The motility of tissue adjacent to that used for histology was measured isometrically in vitro. The increase in the rate of contractions following exposure to physostigmine was greatest for the AGS group, both from the duodenal and from the ileal region. The latency was longest for the AGS group, suggesting that the material from this group had the least number of active cholinergic neurons. The studies with physostigmine thus indicated that the most severe functional damage occurred in cases of AGS. These findings confirm that extensive damage occurs in the enteric neurons in equine grass sickness. There was good correlation between the functional cholinergic responses and the extent of neuronal degeneration. PMID- 9345717 TI - Nephrotoxicity in goats caused by dosing with a water extract from the stems of Narthecium ossifragum plants. AB - Seven goats were given a single dose of an aqueous extract derived from 30 g (wet weight) of Narthecium ossifragum per kg liveweight. Their serum creatinine and urea concentrations increased to day 5 but then fell to normal by day 10. Serum magnesium increased to day 4 and decreased to normal by day 9. Their serum calcium concentration was lower than normal on days 4, 5 and 6. Histopathological examination of the kidneys of goats killed or found dead 2, 4, 6, 8, 11 or 16 days after dosing revealed tubular epithelial cell degeneration and necrosis. Regeneration of the tubular epithelium and signs of interstitial fibroplast proliferation and fibrosis could be seen in animals killed on days 8, 11, 16 and 42. No signs of liver damage were observed in 3 goats dosed with the insoluble plant material from 40 g (wet weight) Narthecium ossifragum per kg liveweight. The total dose was divided into three doses, which were given intraruminally within 7 h. The activities of aspartate aminotransferase, gamma glutamyltransferase and glutamate dehydrogenase remained within the normal range in all 10 goats after dosing. PMID- 9345720 TI - Validation of microsatellite markers for routine horse parentage testing. AB - A parallel testing of 4803 routine Quarter Horse parentage cases, using 15 loci of blood group and protein polymorphisms (blood typing) and 11 loci of dinucleotide repeat microsatellites (DNA typing), validated DNA markers for horse pedigree verification. For the 26 loci, taken together, the theoretical effectiveness of detecting incorrect parentage was 99.999%, making it extremely unlikely that false parentage would fail to be recognized. The tests identified incorrect parentage assignment for 95 offspring (2% of cases). Despite fewer loci, DNA typing was as effective as blood typing and, in parentage exclusion cases, provided more systems to substantiate the genetic incompatibility. Five offspring presented potential genetic incompatibilities with their parents in only a single microsatellite system, but the parentage exclusions could not be confirmed with discordant results at additional loci. Two of these five incompatibilities could be explained as consequences of a null allele and three as fragment size increases or decreases (putative mutations). Provided that an exclusion assignment was based on at least two systems of genetic incompatibility, such rare genetic events did not lead to false exclusions. Notwithstanding the near 100% effectiveness estimations for either typing panel alone to identify incorrect parentage, this validation test showed an actual effectiveness of 97.3% for blood typing and 98.2% for DNA typing. The DNA-based test, however, may feasibly achieve higher efficacy than reported here by adding selected systems to the parentage test panel. PMID- 9345719 TI - The effect of intravenous administration of WEB 2086 on PAF-induced platelet aggregation in healthy Friesian calves. AB - The in vivo ability of the specific PAF-antagonist WEB 2086, a thienotriazolodiazepine, to inhibit platelet-activating factor (PAF) in cattle was investigated by in vitro determination of platelet aggregation curves. WEB 2086 was infused intravenously into a group of 5 healthy male Friesian calves in a dose of 3 mg/kg over 1 min. The resultant inhibition peaked between 30 min and 1 h after administration of WEB 2086. The inhibition was significantly reduced after 3 h and became non-significant after 6 h, but maximal pre-treatment aggregation had not been restored by 24 h after the injection of WEB 2086. These results confirm previous results obtained in vitro and suggest that WEB 2086 is a potent antagonist of PAF activity in calves. They also suggest that further clinical studies with WEB 2086 in cattle are desirable. PMID- 9345721 TI - Genetic mapping of five human chromosome 4 orthologues to bovine chromosomes 6 and 17. AB - Five loci that map to human chromosome 4 (HSA4) were selected to expand the bovine comparative linkage map. Loci included b-casein (CSN2), basic fibroblast growth factor (FGF2), immunoglobulin J chain (IGJ), interleukin 2 (IL2) and microsomal triglyceride transfer protein (MTTP). Polymorphisms for each locus were identified by either polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or single-strand conformational polymorphism (SSCP) analysis. The bovine genes for CSN2, IGJ and MTTP were mapped by linkage analysis to chromosome 6; FGF2 and IL2 mapped to chromosome 17. These data refine a position of chromosomal evolution to a small region between FGF2 and the previously mapped complement I factor (IF). PMID- 9345722 TI - Molecular characterization of major histocompatibility complex (B) haplotypes in broiler chickens. AB - In Leghorn (laying) chickens, susceptibility to a number of infectious diseases is strongly associated with the major histocompatibility (B) complex. Nucleotide sequence data have been published for six class I (B-F) alleles and for class II (B-L beta) alleles or isotypes from 17 Leghorn haplotypes. It is not known if classical B-L or B-F alleles in broilers are identical, at the sequence level, to any Leghorn alleles. This report describes molecular and immunogenetic characterization of two haplotypes from commercial broiler breeder chickens that were originally identified by serology as a single haplotype, but were differentiated serologically in the present work. The two haplotypes, designated BA4 and BA4variant, shared identical B-G restriction fragment length polymorphism patterns, but differed in one B-L beta fragment that cosegregated with the serological B haplotype. Furthermore, the nucleotide sequences of the highly variable exons of an expressed B-L beta II family gene and B-F gene from the two haplotypes were markedly different from each other. Both the B-L beta II family and B-F gene sequences from the BA4 haplotype were identical to the sequences obtained from the reference B21 haplotype in Leghorns; however, in the BA4 haplotype the B-L beta 21 and B-F21 alleles were in linkage with B-G alleles that were not G21. The nucleotide sequences from BA4variant were unique among the reported chicken B-L beta II family and B-F alleles. PMID- 9345723 TI - Linkage of the gene for equine combined immunodeficiency disease to microsatellite markers HTG8 and HTG4; synteny and FISH mapping to ECA9. AB - Equine combined immunodeficiency disease (CID) is caused by homozygosity for an autosomal recessive gene. To identify linked markers for the disease, we studied a family segregating for the equine CID gene. A stallion and 19 of his CID affected offspring were tested for marker segregation at 23 microsatellite DNA loci. His CID-affected offspring inherited only one of his two alleles at the HTG8 and HTG4 loci, namely HTG8-186 and HTG4-124, respectively. Lod scores for linkage to the CID gene using a theta of 0.01 were 5.34 for HTG8 and 2.37 for HTG4. The apparent genotypes also suggested linkage disequilibrium between the HTG8-186 allele and the gene for CID. The gene for the DNA protein kinase catalytic subunit (DNA-PK) was recently suggested as a candidate gene for equine CID. A defect of this gene causes a disease in mice that is similar to equine CID. Therefore, we investigated whether this gene might be associated with the microsatellite markers. Analysis of a somatic cell hybrid panel demonstrated synteny of DNA-PK with HTG4 and HTG8 (Kentucky Synteny Group 3). Fluorescence in situ hybridization (FISH) studies demonstrated that DNA-PK is located on horse chromosome ECA9p12. This work supports the hypothesis of DNA-PK as the probable cause of equine CID. PMID- 9345724 TI - Extensive genomic conservation of cattle microsatellite heterozygosity in sheep. AB - We report the evaluation of 1036 bovine microsatellite primer pairs for their suitability as linkage markers in sheep. Approximately 58% (605/1036) of bovine primer pairs amplified a locus in sheep. Sixty-seven per cent (409/605) of amplified loci were detected as polymorphic. Marker heterozygosity, allele number and range of allele sizes were significantly lower in sheep than cattle sampled in this study. However, median fragment size was similar. These data suggest that high-resolution comparative linkage maps between closely related species can be constructed relatively efficiently. PMID- 9345725 TI - Random amplified polymorphic DNA markers in crosses between inbred lines of Rhode Island red and White Leghorn chickens. AB - Reciprocal crosses and backcrosses were conducted between inbred Rhode Island Red and White Leghorn chickens differentiated for egg production and egg quality traits. Random amplified polymorphic DNA (RAPD) markers distinguishing inbred lines were detected. Twenty-two polymorphic bands were found from screening 120 single 10-mer random primers of which two were consistent with sex-linked markers. Of 90 pairwise two-point linkage analyses completed for the autosomal markers, four close linkages (8.2 cM to 14.9 cM) were significantly different from zero. PMID- 9345726 TI - Characterization of the erythrocyte superoxide dismutase allozymes in the deer Cervus elaphus. AB - The cDNA sequences for Cu,Zn superoxide dismutase from two Cervus elaphus subspecies, North American wapiti and European red deer, were determined. The derived amino acid sequences showed two differences: residue 8 was Leu in wapiti and Met in red deer and residue 25 was His in wapiti and Asn in red deer. The extra positive charge at position 25 in the wapiti isoform accounted for its greater mobility towards the cathode during non-denaturing electrophoresis, a procedure widely used in the genetic analysis of deer. There was no difference in specific activity between the two Cu,Zn superoxide dismutase isoforms, but the wapiti isoform was slightly more susceptible to heat denaturation. PMID- 9345727 TI - Polymorphic microsatellite loci in the European rabbit (Oryctolagus cuniculus) are also amplified in other lagomorph species. AB - Six polymorphic microsatellite markers developed for the European wild rabbit (Oryctolagus cuniculus) were amplified with 20 other species of lagomorphs, representing both commercially important species and species important from a conservation perspective. Successful amplification of a number of these loci has provided an important set of new molecular markers for this mammalian order. PMID- 9345728 TI - Genetic relationship between equine apolipoproteins A4 and A1. AB - Genetic polymorphism of equine apolipoprotein (APO) A4 was investigated using two dimensional electrophoresis in four horse breeds, including Japanese native horses. A linkage relationship between the equine APOA4 and APOA1 structural loci was assumed from the segregation data of these loci in one family line of the Japanese Hokkaido native breed. PMID- 9345729 TI - A PCR polymorphism detected in the goat alpha s2-casein gene. PMID- 9345730 TI - Syntenic assignment of thrombomodulin (THBD) and phosphatidylinositol-specific phospholipase C (PLC-II) to bovine chromosome 13. PMID- 9345732 TI - Dinucleotide repeat polymorphisms at seven anonymous microsatellite loci cloned from the European harbour seal (Phoca vitulina vitulina). PMID- 9345731 TI - Equine dinucleotide repeat loci LEX034-LEX048. PMID- 9345733 TI - Rapid genotyping of a pig minisatellite using long PCR. PMID- 9345734 TI - Nine novel chicken microsatellite loci and their utility in other Galliformes. PMID- 9345735 TI - Three newly detected alloantigens in the U blood group system of horses. PMID- 9345736 TI - A polymorphic dinucleotide microsatellite in rainbow trout (Onchorhynchus mykiss): OmDIAS1. PMID- 9345739 TI - Chicken solute carrier family 4 anion exchanger member 1 (SLC4A1): microsatellite polymorphism and mapping. PMID- 9345738 TI - Linkage mapping of a bovine alpha-lactalbumin pseudogene (LALBAps) using a LINE associated polymorphism. PMID- 9345737 TI - PCR-based diagnostic tests for the endogenous retroviral elements ev-B1 and ev-B5 of chickens. PMID- 9345740 TI - A pentanucleotide repeat polymorphism maps progesterone receptor (PGR) to chicken chromosome 1. PMID- 9345741 TI - A HpaII polymorphism in goat mitochondrial DNA. PMID- 9345742 TI - Two bovine polymorphic microsatellite loci (PZ251 and PZB2F). PMID- 9345743 TI - A polymorphic Y chromosome microsatellite locus in cattle. PMID- 9345744 TI - BanII RFLP at the apolipoprotein E (APOE) locus in baboons. PMID- 9345745 TI - PCR/RFLP marker in the canine transducin-gamma gene (GNGT1). PMID- 9345746 TI - A PstI RFLP at the porcine POU domain class 1 transcription factor 1 (POU1F1) gene. PMID- 9345747 TI - Detection of intronic sequence in the bovine ITGB2 gene. PMID- 9345748 TI - Detection of a DdeI PCR RFLP within intron 1 of the porcine MYOD1 (MYF3) locus. PMID- 9345749 TI - An EcoRI polymorphism at the beta-nerve growth factor (NGFB) locus in cattle. PMID- 9345750 TI - Unusual lactam formation occurring in the synthesis of a biotinylated T-antigen serine derivative. AB - Synthesis of the biotinylated T-antigens, linked to a serine by an alpha (7 alpha) or a beta (7 beta) 2-acetamido-2-deoxy-D-galactoside bond, is described. These derivatives were needed for the detection of a specific endogenous lectin at the surface and/or on the migration pathway of melanoma cells. In the course of the synthesis, an unusual lactam formation was observed with the beta anomer of the azido-disaccharide 5 beta. PMID- 9345751 TI - Synthesis of L-ribofuranosyl C-nucleosides. AB - C-Nucleosides, 4-amino-8-(beta-L-ribofuranosyl)pyrazolo[1, 5-a]-1,3,5-triazine (12) and 4-amino-7-(beta-L-ribofuranosyl)-5H-pyrrolo[3,2-d]pyrimidine (L-9 deazaadenosine, 22), were synthesized from the key intermediate, 3-dimethylamino 2-(2,3-O-isopropylidene-5-O-trityl-L-ribofuranosyl) acrylonitrile (8), which was prepared from L-xylose in 11 steps. PMID- 9345752 TI - A simple access to lactose-derived building blocks required in glycoconjugate synthesis. AB - Lactose was readily transformed into thexyldimethylsilyl (3,4-O-isopropylidene beta-D-galactopyranosyl)-(1-->4)-beta- D-glucopyranoside (5); this compound served as intermediate for the generation of partially O-protected lactose building blocks required in oligosaccharide and glycoconjugate synthesis. Thus, from 5 via per-O-benzoylation, desilylation, trichloroacetimidate formation, glycosylation of the Lemieux spacer, and acid-catalyzed de-O-isopropylidenation methoxycarbonyloctyl (2,6-di-O-benzoyl-beta-D-galactopyranosyl)- (1-->4)-2,3,6 tri-O-benzoyl-beta-D-glucopyranoside (12) was obtained. Regioselective benzoylation of 5 with benzoyl cyanide under various conditions afforded 3-O- (13), 2,3,2'-O- (14), 3,2'-O- (16), and 2,2'-O-unprotected (17) lactoside, respectively. De-O-isopropylidenation of 16 gave thexyldimethylsilyl (6-O-benzoyl beta-D-galactopyranosyl)-(1-->4)-2, 6-di-O-benzoyl-beta-D-glucopyranoside (18), an important 2',3',4'-O-unprotected lactose derivative. Fucosylation of 13 and then de-O-isopropylidenation afforded thexyldimethylsilyl 2,6-di-O-benzoyl-beta-D galactopyranosyl)-(1-->4)-[(3,4-di-O-acetyl-2-O- benzoyl-alpha-L-fucopyranosyl) (1-->3)]-2,6-di-O-benzoyl-beta-D- glucopyranoside (21), an important fucosyllactose building block. PMID- 9345753 TI - Structural elucidation of acidic fungal polysaccharides isolated from the cell wall of genera Cylindrocladium and Calonectria. AB - The structure of acidic fungal polysaccharides, isolated from the cell wall of Cylindrocladium and Calonectria species, has been investigated by chemical analysis, methylation and reductive cleavage analyses, and 1D and 2D 1H and 13C NMR spectroscopy. The polysaccharides have an idealized repeating unit: [formula: see text] linked to a small mannan core (< 5%), with n = 3 for Cylindrocladium penicilloides and C. quinqueseptatum and n = 5 for Calonectria theae, C. crotalariae, and C. colhounii. PMID- 9345754 TI - Anhydro sugar and linkage contributions to circular dichroism of agarose and carrageenan, with conformational implications. AB - The geometry-dependent linkage contributions to the circular dichroism (CD) of agarose and carrageenan were determined by subtracting the monomeric CD from the CD of the polymers. For this purpose, the CD of methyl 3,6-anhydro-alpha-D galactopyranoside, and of other anhydro sugars, was measured. Application of empirical quadrant rules indicates the observed CD to be that expected for the helical conformations of agarose and carrageenan as derived from diffraction data and modeling calculations. The difference in CD sign in the two polymers arises from a translocation of the beta-D-galactose O-2 atom from one quadrant to the neighboring quadrant of the C-5-O-5-C-1 ether chromophore of the preceding anhydro sugar residue, demonstrating the unusually high spatial resolution of conformational analysis that can be achieved with CD under favorable circumstances. PMID- 9345756 TI - Structure of a sulfated xylogalactan from the calcareous red alga Corallina pilulifera P. et R. (Rhodophyta, Corallinaceae). AB - The structure of a sulfated polysaccharide isolated from the calcareous red alga Corallina pilulifera was studied by methylation analysis before and after desulfation or Smith degradation, as well as by 1D and 2D 1H and 13C NMR spectroscopy. The polysaccharide was shown to consist of D-galactose, L galactose, 2-O-methyl-L-galactose, 3-O-methyl-L-galactose, 6-O-methyl-D galactose, D-xylose, and sulfate in a molar ratio of 29:20:5:2:1:20:23. Its agaran-like backbone built up of alternating 3-linked beta-D-galactopyranose and 4-linked alpha-L-galactopyranose residues bears single beta-D-xylopyranosyl substituents at position 6 of beta-D-galactose residues, whereas sulfate and O methyl groups occupy positions 2 and 3 of alpha-L-galactose and position 6 of beta-D-galactose residues. PMID- 9345755 TI - Structural characterisation of galactoglucomannan secreted by suspension-cultured cells of Nicotiana plumbaginifolia. AB - Galactoglucomannan (GGM) from cultures of Nicotiana plumbaginifolia has Man:Glc:Gal:Ara:Xyl in 1.0:1.1:1.0:0.1:0.04 ratio. Linkage analysis contained 4- and 4,6-Manp, 4-Glcp, terminal Galp and 2-Galp, small amounts and terminal Arap and terminal Xylp, and approximately 0.03 mol acetyl per mol of glucosyl residue. Treatment with alpha- and beta-D-galactosidases showed that the majority of the side-chains were either single Galp-alpha-(1-->residues or the disaccharide Galp beta-(1-->2)-Galp-alpha-(1-->linked to O-6 of the 4-Manp residues of the glucomannan backbone. Analysis of the oligosaccharides generated by endo-(1-->4) beta-mannanase digestion confirmed that the GGM comprises a backbone of predominantly alternating-->4)-D-Manp-beta-(1-->and-->4)-D-Glcp-beta-(1-->branch ed at O-6 of 65% of the 4-Manp residues. The major oligosaccharide identified was D-Glcp-beta-(1-->4)-[D-Galp-beta-(1-->2)-D-Galp-alpha-(1-->6)]-D-Man p-beta-(1- >4)-D-Glcp-beta-(1-->4)-[D-Galp-alpha-(1-->6)]-D-Manp -beta-(1-->(27%), and most of the other oligosaccharides produced in significant quantities were based on this structure. PMID- 9345757 TI - Synthesis of lactosyl phosphate diester derivatives of nucleosides. AB - Derivatives of the lactosyl phosphate diester and the lactosyl thiophosphate diester of 1-(beta-D-arabinofuranosyl)cytosine (Ara-C) and 9-(beta-D arabinofuranosyl)adenine (Ara-A) were synthesized by condensation of 3 hydroxypropyl lactoside and a protected Ara-C or Ara-A via H-phosphonate methodology and phosphoramidite methodology, respectively. PMID- 9345759 TI - Enhancement of ColE1-type replication promoted by the Gp alpha initiator protein of bacteriophage P4. AB - Gp alpha, the phage P4 specific replication protein, increases in vitro replication of pCN51, a pBR322 based replicon, by a factor of two. This effect is dependent on DNA polymerase I and requires transcription by host RNA polymerase. Electron microscopic analysis of replicating intermediates indicates that pCN51 replication occurred from the same origin and with the same directionality in the presence and in the absence of Gp alpha. These results reveal that Gp alpha can influence the replication of an heterologous replicon and show that this effect occurs in ColE1-type replicons without altering the normal pattern of initiation. Further analysis of replicating intermediates shows an increase in the average size of the ColE1-type replication 'bubble' obtained in the presence of Gp alpha. It is proposed that Gp alpha interacts with the ColE1 replisome complex at an early replication stage. PMID- 9345760 TI - Brucella Omp2a and Omp2b porins: single channel measurements and topology prediction. AB - Brucella usually carry two highly homologous genes (omp2a and omp2b) for porin like proteins. In several B. abortus biovars the omp2a gene has a large deletion compared to other Brucella omp2's. In this study we have measured Omp2 pore activity in planar bilayers. Omp2b exhibits well-defined trimeric channel activity whilst Omp2a forms monomeric pores of variable size which are smaller than Omp2b. No sequence homology exists between Omp2 and porins of known structure, so hydrophobic moment analysis has been used to model their membrane topology. From this it appears likely that the deletion removes the crucial L3 internal loop. PMID- 9345761 TI - Antibody-mediated selection of a Mycoplasma gallisepticum phenotype expressing variable proteins. AB - A variant phenotype of Mycoplasma gallisepticum S6 was isolated from an in vitro antibody-culture system utilizing metabolism-inhibiting antibodies against the 64 kDa lipoprotein (LP64). M. gallisepticum populations grown in medium alone or medium containing normal rabbit serum maintained expression of the parental phenotype. This paper describes the identification of proteins which undergo variable expression. Several of these were integral membrane proteins, with estimated molecular masses of 91, 43, 41, 38, 37, and 18 kDa, which were expressed in the variant phenotype but not in the parental phenotype. Three proteins (LP64, p63 and p47) were expressed in the parental phenotype, but not in the variant phenotype. The data suggest that the interaction of specific immunoglobulins with target epitopes resulted in the selection of a subpopulation of organisms expressing an alternative array of membrane proteins which, lacking the target epitopes, was able to escape the metabolism-inhibiting effects of the specific antibodies. PMID- 9345762 TI - Construction of a double hha hns mutant of Escherichia coli: effect on DNA supercoiling and alpha-haemolysin production. AB - A double hha hns Escherichia coli mutant was constructed. The effect of the single hns mutation and of the double hha hns mutation on the expression of the alpha-haemolysin determinant of plasmid pANN202-312 was assessed. Whereas the hns mutant moderately increased expression of the toxin, the double hha hns mutant strongly enhanced transcription of the hly operon and hence expression of the toxin. This suggests that both Hha and H-NS proteins participate in the modulation of the expression of the toxin. The enhancement of haemolysin expression in the double mutant could not be correlated to a global alteration of DNA topology: DNA preparations of a reporter plasmid isolated from this mutant gave a topoisomer distribution similar to that of the parental strain. PMID- 9345763 TI - The C-terminal ligand-binding domain of Clostridium difficile toxin A (TcdA) abrogates TcdA-specific binding to cells and prevents mouse lethality. AB - We have investigated the ability of a recombinant protein (REP231), derived from Clostridium difficile toxin A C-terminal domain, to protect against toxin A (TcdA) intoxication in vitro and in vivo. REP231 was cloned, expressed and purified by thyroglobulin affinity chromatography, and demonstrated identical binding properties to TcdA. Immunofluorescence experiments and in vitro cytotoxicity assays using mouse teratocarcinoma cells F9 showed that specific binding of TcdA to F9 cells through its C-terminal domain is essential for producing cytotoxic effects. TcdA binding and cytotoxicity was inhibited by REP231 and a monoclonal antibody directed against the C-terminal domain. Toxin B did not bind to F9 cells and was consequently inactive in cytotoxicity assays. Inhibition studies with lectins and a Le(x)-specific antibody supported earlier findings that a terminal galactose is part of the bound saccharide but excluded Le(x) as a receptor for TcdA. Mice immunised with REP231 were protected against a threefold lethal dose of TcdA. Thus, REP231 appeared to be a suitable candidate to develop an alternative therapeutic agent, which is able to neutralise carbohydrate-mediated TcdA binding and might act as a vaccine. PMID- 9345764 TI - Arrangement of the vanA gene cluster in enterococci of different ecological origin. AB - Glycopeptide-resistant enterococci (vanA) isolated from infections in humans, from non-hospitalized humans, from sewage, from animal feces and from meat products in Germany (20 Enterococcus faecium and one Enterococcus hirae) were investigated for the arrangement of the genes in the vanA gene cluster by means of overlapping PCR with five primer pairs. In 20 of these strains, the vanA gene clusters were uniform which suggests a horizontal spread among different ecosystems. In one clinical isolate a rearrangement was detected in the vanY-vanZ region. PMID- 9345765 TI - Evidence of rare codon clusters within Escherichia coli coding regions. AB - It is known that there is a high occurrence of rare codons at the start of coding region. Here it is shown that although the remainder of the gene is likely to contain a relatively low number of rare codons, rare and non-rare codons do not form a random sequence. It is apparent that throughout the coding region there is a higher than expected number of rare codon clusters. For example once a rare codon has occurred there is a greater chance than expected of the next six codons containing another rare codon. This non-random distribution implies that rare codons may have an as yet unidentified biological role. PMID- 9345766 TI - Blepharismin produced by a protozoan Blepharisma functions as an antibiotic effective against methicillin-resistant Staphylococcus aureus. AB - A ciliated protozoan, Blepharisma japonicum, produces a photosensitive red pigment, blepharismin (BLR). This study showed that the pigment inhibits the growth of Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA) resistant to arbekacin (ABK), which is the most effective aminoglycoside antibiotic against MRSA and used world wide. Although the minimum inhibitory concentration (MIC) of BLR to the ABK-resistant MRSA strain was 6.25 micrograms/ml in dark, it was decreased to 1.25 micrograms/ml by irradiation with white light of 65 W/m2 for 30 min, suggesting that the antibacterial activity of BLR is photoactivated. Our findings suggested that the antibacterial activity of BLR in dark is due to inhibition of protein synthesis. In addition, we found that BLR is bactericidal and enhances synergistically the antibacterial activity of ABK. PMID- 9345767 TI - Function and regulation of the acid and neutral trehalases of Mucor rouxii. AB - Two different trehalose-hydrolysing activities, known as acid or non-regulatory trehalases, and neutral or regulatory trehalases, have been recognised in a number of fungal species. The true role of these apparently redundant hydrolases remained obscure for many years. However, recent evidence suggests that neutral trehalases would be specialised in the mobilisation of cytosolic trehalose, while acid trehalases would only hydrolyse extracellular trehalose. Results obtained with Mucor rouxii, a Zygomycete initially thought to possess only neutral trehalase activity, reinforced this hypothesis. M. rouxii grows efficiently in trehalose as the sole carbon source. Trehalose-grown or carbon-starved cells exhibit a high trehalase activity of optimum pH 4.5, bound to the external surface of the cell wall, in contrast with the neutral (pH 6.5) trehalase, which occurs in the cytosol. Other differences between the neutral and the acid trehalases are the temperature optimum (35 degrees C and 45 degrees C, respectively) and thermal stability (half-life of 2.5 min and 12 min at 45 degrees C, respectively). The neutral trehalase, but not the acid trehalase, is activated in vitro by cAMP-dependent phosphorylation, stimulated by Ca2+, and inhibited by EDTA. It shows maximal activity at germination and decreases as growth proceeds. In contrast the activity of the acid trehalase is totally repressed in glucose-grown cultures and increases upon exhaustion of the carbon source, and is strongly induced by extracellular trehalose. PMID- 9345768 TI - Cloning and sequencing of a second laccase gene from the white-rot fungus Coriolus versicolor. AB - A second laccase gene, CVLG1, was isolated from Coriolus versicolor. CVLGI encodes a precursor protein of 526 amino acids which contains a 23-amino acid signal sequence, and the coding region is interrupted by 11 introns. The number of potential N-glycosylation sites in this product is 12 and the greatest among that of polyporales laccases. Moreover, this protein shares about 70% homology with other polyporales laccases. Genomic Southern analysis showed that C. versicolor laccases are encoded by more than four genes including CVLG1 and a transposed allele of this gene. PMID- 9345769 TI - Evidence for natural transformation of Bacillus subtilis in foodstuffs. AB - The effect of foodstuffs on the natural transformation of Bacillus subtilis was investigated. As examples of complex food matrices milk with various fat contents as well as chocolate milk were used. The frequencies of transformation varied with the fat content and ranged between 3.8 X 10(-4) and 1.4 X 10(-3). Highest frequencies of about 3 X 10(-3) were observed in chocolate milk with 1.5% fat. Development of competence was observed in chocolate milk, resulting in maximal transformation frequencies upon incubation for 10-12 h at 37 degrees C. PMID- 9345770 TI - Cloning and characterization of genes encoding an enzyme which oxidizes dimethyl sulfide in Acinetobacter sp. strain 20B. AB - Acinetobacter sp. strain 20B was isolated based on the ability to utilize dimethyl sulfide as the sole sulfur source. Since strain 20B oxidized indole as well as dimethyl sulfide, indigo production by recombinant Escherichia coli clones carrying Acinetobacter DNA was used as a selection for cloning genes encoding dimethyl sulfide oxidation genes. The gene encoding an indole-oxidizing enzyme was also found to oxidize dimethyl sulfide. The dimethyl sulfide-oxidizing enzyme genes consisted of six open reading flames designated dsoABCDEF. The deduced amino acid sequences of dsoABCDEF were homologous with those of the multicomponent phenol hydroxylases. DsoABCDEF oxidized dimethyl sulfide to dimethyl sulfoxide, and dimethyl sulfoxide to dimethyl sulfone. PMID- 9345771 TI - Identification and characterization of a Treponema pallidum subsp. pallidum gene encoding a DNA adenine methyltransferase. AB - The nucleotide sequence of a DNA adenine methyltransferase gene (dam) from Treponema pallidum has been determined. Southern blot analysis of T. pallidum chromosomal DNA indicated that this gene is present as a single copy. The dam gene encodes a 303 amino acid protein whose deduced sequence has significant homology with DNA (N6-adenine) methyltransferases. T. pallidum Dam can be assigned to group alpha DNA amino methyltransferases based on the order of nine conserved motifs that are present in the protein. Digests of T. pallidum chromosomal DNA performed with isoschizomer restriction endonucleases (Sau3AI, DpnI, and MboI) confirmed the presence of methylated adenine residues in GATC sequences (Dam+ phenotype). PMID- 9345772 TI - A double-stranded RNA mycovirus in Botrytis cinerea. AB - In wild-type Botrytis cinerea CVg25 strain we have detected the presence of extrachromosomal genetic elements corresponding to double-stranded RNA molecules. These genetic elements have been designated L, M1 and M2 with molecular sizes of 8.3, 2.0 and 1.4 kb, respectively. The visualization by electron microscopy of mycelium ultrathin sections from B. cinerea CVg25 showed the presence of isometric virus-like particles of about 40 nm in diameter. Linear sucrose gradient centrifugation of mycelium-free extracts was done to determine if the double-stranded RNAs were associated with virus-like particles. The gradient profile obtained at 260 and 280 nm revealed a major peak that was analyzed by both agarose-gel electrophoresis and electron microscopy. It was observed that only the L-double-stranded RNA molecule copurified with isometric virus-like particles. These virus-like particles had a similar morphology and size as those detected by electron microscopy in the mycelium sections. These results suggest that only the L-double-stranded RNA would be encapsidated. PMID- 9345773 TI - A universal post-embedding protocol for immunogold labelling of osmium-fixed, epoxy resin-embedded tissue. AB - The authors have found that double etching of epoxy-embedded, ultrathin sections with 3% sodium metaperiodate rendered epitope expression comparable to that obtained with either saturated or half-saturated sodium metaperiodate solutions. In contrast to either of the two more concentrated oxidizing solutions, double etching with 3% sodium metaperiodate neither bleached the specimens nor generated holes in the plastic resin. PMID- 9345774 TI - Atomic force microscopy of collagen molecules. Surface morphology of segment-long spacing (SLS) crystallites of collagen. AB - Three-dimensional structures of laterally aggregated bundles of collagen molecules, segment-long-spacing (SLS) crystallites, were imaged by atomic force microscopy (AFM) in atmosphere. The overall image of the type I collagen SLS in a height-amplified mode was semi-cylindrical, approximately 300 nm in length, with two bands of elevation near both N- and C- ends of the molecule. Its 'cross sectional' profile (across the molecular axis) was a smooth arch. The 'axial' profile (along the molecular axis) had two prominent peaks approximately 250 nm apart, corresponding to the two bands of elevation. There were several minor peaks between these two prominent peaks. The elevation near both ends may be due to the presence of covalently bound sugars near both N- and C- ends of the helical part of type I collagen alpha chains. The AFM images of SLS presented here indicate that the type I collagen molecule is not uniform in diameter and has two bulged parts within its triple helix. PMID- 9345775 TI - Particle size does not affect the rate of intracellular routing for ligands internalized by non-adsorptive pinocytosis. AB - The rate at which three different species of a fluid phase marker (horseradish peroxidase) reached the lysosomal compartment of mouse LM fibroblasts was compared. Initial observations suggested that the rate was a function of particle size. Adjustment of the concentrations of the various species of HRP to equal numbers of physical particles negated the apparent affect. Thus, particle size does not affect the intracellular routing of a ligand entering fibroblasts by non adsorptive pinocytosis. PMID- 9345776 TI - Electron microscopic observations on the normal development of Trichinella spiralis from muscle larvae to adult worms in BALB/c mice with emphasis on the body wall, genital organs and gastrointestinal organs. AB - Ultrastructural changes during the development of Trichinella spiralis from muscle larvae to adult worms are described with emphasis on the body wall, genital organs and gastrointestinal organs. The cuticle of the body wall and hindgut underwent the first molting at 14 h post-infection (PI). The esophagus cuticle never molted. The hypodermal gland was formed in the lateral cords by 30 h PI. The genital primordium developed to either male or female genitals, accomplishing their sexual maturity and fertilization by 3 days PI. Glycogen disappeared during the development. PMID- 9345777 TI - Isolation of intact collagen fibrils from healing ligament. AB - The inability to isolate intact collagen fibrils has limited the study of their growth and structure. Although intact fibrils have been isolated from echinoderms and from embryonic chick tissues, no method has previously succeeded in isolating intact collagen fibrils from a postfoetal vertebrate tissue. Having previously observed that gentamicin weakens rat tail tendon, we hypothesized that gentamicin may weaken interfibrillar bonds and that intact collagen fibrils might be isolated from tissue treated with gentamicin. In this study medial collateral knee ligaments of Sprague Dawley rats were transected and then harvested 24, 48 or 96 h postoperatively. These specimens were placed in gentamicin or phosphate buffered saline for 72 h, vortexed for 1 h, incubated in gentamicin for an additional 24 h, and vortexed again for 1 h. Negatively stained specimens were examined with a transmission electron microscope. The phosphate-buffered saline specimens yielded only broken fibrils. The gentamicin specimens yielded both broken and intact fibrils. The latter and tapering ends and consisted of molecules orientated such that their amino termini pointed toward the tip and their carboxy termini pointed toward a short central region where the molecular polarity reversed. PMID- 9345779 TI - Shimming a high-resolution MAS probe. AB - A systematic and efficient approach to shimming a high-resolution, magic angle sample spinning probe is introduced. The method takes into account the different symmetries of the normal shim coils and the MAS experiment. PMID- 9345778 TI - Rotational correlation times of internuclear vectors in a DNA duplex with G-A mismatch determined in aqueous solution by complete relaxation matrix analysis of off-resonance ROESY (O-ROESY) spectra. AB - Complete relaxation matrix analysis of the off-resonance ROESY (O-ROESY) spectra is presented and demonstrated for a synthetic DNA duplex with two G-A mismatches, d(GCTGTC-GAAAGC)2, in solution. The internuclear distance and the rotational correlation time of the internuclear vector could be garnered simultaneously using complete relaxation matrix analysis of O-ROESY, by which spin diffusion effects could be accommodated. Correlation times in the terminal and the mismatched regions were significantly reduced compared to those in other regions, indicating the conformational flexibility of the mismatched pair. The average structure obtained by restrained molecular dynamics simulation with inclusion of variations of the rotational correlation times also indicated a general tendency of the mismatched and contiguous bases to flip to the outside of the double strand. Off-resonance ROESY combined with the complete relaxation matrix analysis method may offer an alternative way to investigate the structures and dynamics of biological macromolecules. PMID- 9345780 TI - Photoreactions of ruthenium(II) and osmium(II) complexes with deoxyribonucleic acid (DNA). AB - The design of Ru(II) and Os(II) complexes which are photoreactive with deoxyribonucleic acid (DNA) represents one of the main targets for the development of novel molecular tools for the study of DNA and, in the future, for the production of new, metal-based, anti-tumor drugs. In this review, we explain how it is possible to make a complex photoreactive with nucleobases and nucleic acids. According to the photophysical behaviour of the Ru(II) compounds, two types of photochemistry are expected: (1) photosubstitution of a ligand by a nucleobase and another monodentate ligand, which takes place from the triplet, metal-centred (3MC) state; this state is populated thermally from the lowest lying triplet metal to ligand charge transfer (3MLCT) state; (2) photoreaction from the 3MLCT state, corresponding to photoredox processes with DNA bases. The two photoreactivities are in competition. By modulating appropriately the redox properties of the 3MLCT state, an electron transfer process from the base to the excited complex takes place, and is directly correlated with DNA cleavage or the formation of an adduct of the complex to DNA. In this adduct, guanine is linked by N2 to the alpha-position of a non-chelating nitrogen of the polyazaaromatic ligand without destruction of the complex. Different strategies are explained which increase the affinity of the complexes for DNA and direct the complex photoreactivity to sites of special DNA topology or targeted sequences of bases. Moreover, the replacement of the Ru(II) ion by the Os(II) ion in the photoreactive complexes leads to an increased specificity of photoreaction. Indeed, only one type of photoreactivity (from the 3MLCT state) is present for the Os(II) complexes because the 3MC state is too high in energy to be populated at room temperature. PMID- 9345781 TI - Effects of fractionated 5-aminolevulinic acid administration on tissue levels of protoporphyrin in vivo. AB - The photodynamic therapy (PDT) of malignant tissues can be achieved via the administration of 5-aminolevulinic acid (ALA), which is naturally converted to the photoreactive substance protoporphyrin IX (PP). This study compares bolus with fractionated ALA dosing in order to determine whether one of these methods results in a higher tissue concentration of PP. Mice bearing a subcutaneously implanted colon-26 tumor were treated with ALA (200 mg kg-1), given intravenously either as a single bolus or as three equally divided doses at 50 min intervals. Tissue samples of tumor, kidney, skin, liver, skeletal muscle, colon and plasma were obtained 2, 3, 4 and 6 h later for the analysis of PP concentrations. Fractionated dosing results in significantly higher concentrations of PP at 4 and 6 h for kidney, 3 and 6 h for skin, 3 h for colon and 6 h for liver. In contrast, fractionated dosing has no significant effect on the PP concentrations of muscle and plasma. Fractionated dosing results in a significantly greater PP concentration in the tumor at 3 h relative to that observed for the bolus dose. However, from a consideration of the time of PP measurement, it is concluded that fractionated dosing may not cause a significant increase in the PP concentration in colon-26 tumors relative to that observed for the bolus dose. PMID- 9345782 TI - Fluorescent lipid probes 12-AS and TMA-DPH report on selective, purinergically induced fluidity changes in plasma membranes of lymphoid cells. AB - The effect of extracellular ATP (ATPex) on the anisotropy of 1-[4 (trimethylamino) phenyl]-6-phenyl-hexa-3,5 triene (TMA-DPH) and 12-anthroyloxi stearic acid (12-AS) fluorescence was investigated in Balb/C mouse thymocytes and in JY human lymphoblasts. These cells have been shown recently to be sensitive and resistant to ATPex, respectively, in terms of cellular responses. Extracellular ATP (1 mM) induced a time-dependent elevation in the emission anisotropy of both probes (indicating an increased lipid packing density) in the plasma membrane of thymocytes. The maximal effect, at 37 degrees C, was observed between 20 and 60 min after ATPex administration, and followed by a gradual decrease of fluorescence anisotropy at longer times (60-180 min). ATPex did not change membrane fluidity of thymocytes below the phase transition temperature (at 18 degrees C). Oxidized ATP (oATP), a selective antagonist of P2z purinoreceptors, blocked the ATPex-induced decrease in membrane fluidity. Low ATPex concentrations (100-300 microM)--which are known to induce distinct signals (changes in membrane potential and intracellular Ph)--slightly fluidized the plasma membrane of thymocytes. This effect was partially blocked by quinine, a blocker of Ca(2+)-activated K+ channels. Neither 12-AS nor TMA-DPH showed any change in their emission anisotropy upon ATPex-treatment in the plasma membrane of the resistant human JY lymphoblast cells. No other signalling event (membrane potential change, Ca2+ response) is elicited by ATPex in this cell line. These data suggest that the changes in the membrane fluidity are likely consequences of specific, purinoreceptor-mediated signalling events, such as hyper-or depolarization of the plasma membrane or Ca2+ influx. These signals may induce changes in the conformation or lateral organization of membrane proteins, perturbing protein-lipid interactions, as well. PMID- 9345783 TI - Single strand breaks and mutagenesis in yeast induced by photodynamic treatment with chloroaluminum phthalocyanine. AB - Photodynamic treatment of the yeast Kluyveromyces marxianus with the sensitizer aluminum phthalocyanine results in loss of clonogenicity. In this paper the effect of this treatment on DNA of this yeast was investigated by searching for single strand breaks and forward mutations. Using the alkaline step elution technique it was found that illumination of the yeast in the presence of aluminum phthalocyanine resulted in an increase in single strand breaks. These could, partially, be repaired by post-incubating illuminated cells in growth medium. At comparable survival levels, photodynamic treatment with aluminum phthalocyanine induced fewer single strand breaks than X-ray treatment. By using a medium containing 5-fluoroorotic acid, mutants in the uracil biosynthetic pathway were selected. Photodynamic treatment resulted in a light dose dependent increase of the mutation frequency. The observed mutagenicity of photodynamic treatment of the yeast with phthalocyanine was lower than the mutagenicity of UVC and X-ray treatment at equal colony forming capacity, indicating that photodynamic treatment is the least mutagenic of those treatments. It is concluded that photodynamic treatment of K. marxianus results in DNA damage. Saccharomyces cerevisiae rad14 and rad52 mutants were used to determine the effect of the nucleotide excision repair and recombinational repair pathways, respectively, on survival after photodynamic treatment. Our data indicate that DNA damage is not the main determinant for cell killing by photodynamic treatment and that the type of damage induced is apparently not subject to RAD14- or RAD52 controlled repair. PMID- 9345784 TI - Pharmacokinetic and phototherapeutic properties of axially substituted Si(IV) tetradibenzobarreleno-octabutoxyphthalocyanines. AB - Three Si(IV)-tetradibenzobarreleno-octabutoxyphthalocyanines (TDiBOPcs) bearing different axial ligands on the metal ion were studied for their tumour-localizing and-photosensitizing properties after i.v. injection via a Cremophor emulsion (0.35 mumol kg-1 b.w.) to Balb/c mice bearing an intramuscularly implanted MS-2 fibrosarcoma. In all cases, the maximum tumour accumulation of the photosensitizer (0.8-1.9 nmol g-1 of tissue) was found at 24 h after injection. The efficiency and selectivity of tumour targeting appeared to be dependent on the nature of the axial ligands; optimal values of these parameters were obtained in the case of the bis(trihexyl-siloxy)-substituted Si(IV)-TDiBOPc, which gave a 7-9 tumour/muscle ratio of phthalocyanine concentration at 24-48 h after injection. The extent of tumour response to PDT treatment was correlated with the concentration of the photosensitizer in the tumour tissue: upon 740 nm irradiation (180 mW cm-2, 200 J cm-2) at 48 h after injection of 0.35 mumol kg-1 of Si(IV)-TDiBOPc-C6H13, the tumour growth exhibited a delay of about 7 days. PMID- 9345785 TI - Spectral variations of laser-induced tissue emission during in vivo detection of malignancies in the female genital tract. AB - A laser-based fluorescence-guided biopsy system has been developed for the screening and early detection of malignancies in the female inner/outer genital tract. Fluorescence spectra were recorded during in vivo exposure of normal and malignant tissue to He-Cd laser (442 nm) radiation. Spectral distribution of tissue natural fluorescence allowed for the development of simple algorithms, based on spectral intensity variations. A subsequent index of discrimination between normal and various malignant tissues has been calculated. This study focuses on the variability of the experimental data and the possible sources of spectral variations. These results suggest that monitoring of this index during colposcopy could enhance selective detection of the malignant tissue, reducing the risk of leaving pathologic tissue untreated during standard exploratory surgical procedures. PMID- 9345786 TI - Biophoton emission of the human body. PMID- 9345787 TI - Sensitivity of aerobic spore-forming species of the genus Bacillus to the pteridine compound O129. AB - Discs containing the pteridine compound O129 at various concentrations may be useful in differentiating the following closely related species belonging to the genus Bacillus, viz. B. subtilis, B. licheniformis and B. pumilus; B. polymyxa and B. macerans; together with B. cereus and its subspecies B. cereus var. mycoides. At concentrations of 10 micrograms O129, the ATCC type strain of B. subtilis was resistant whereas B. licheniformis and B. pumilus were sensitive. However, B. subtilis was sensitive to O129 at 150 micrograms. Similar reactions differentiated the ATCC type strains of B. polymyxa and B. macerans. The ATCC type strain of B. cereus was resistant to O129 at both 150 micrograms and 10 micrograms, but its subspecies B. cereus var. mycoides (ATCC 28) was partially sensitive at 150 micrograms concentration. When clinical isolates of B. cereus var. mycoides were subsequently tested, this partial sensitivity was found to be a variable characteristic. PMID- 9345788 TI - Mycoflora and mycotoxins in adzuki and mung beans produced in Ontario, Canada. AB - This is the first report on the detection of fumonisin B1 (FB1) in Fusarium infected adzuki bean (Phaseolus angularis) and mung bean (Phaseolus aureus). The infected beans had either a mouldy appearance or a distinct discoloration. Seed coats of infected adzuki beans changed from dark red to light red and those of mung beans from green to dark or brownish green. Fusarium spp. isolated from mouldy and discoloured beans included F. avenaceum, F. culmorum, F. equiseti, F. graminearum, F. moniliforme, F. oxysporum, F. solani, F. sporotrichoides and a few unidentified species. Healthy beans without any apparent discoloration and diseased beans with discoloration and mouldy appearance were analysed for mycotoxins. Diacetoxyscripenol, deoxynivalenol and T-2 toxin (T-2) were not detected in either healthy or discoloured adzuki and mung bean samples by thin layer chromatography (TLC). FB1 was detected by TLC in discoloured adzuki and mung bean samples but not in the healthy samples. TLC results were confirmed by high performance liquid chromatography (HPLC). The quantification of FB1 by HPLC revealed that discoloured adzuki and mung bean samples contained 261 +/- 43.8, and 230 +/- 21.6 micrograms g-1 of FB1, respectively. This investigation emphasizes the need for more detailed research in dealing with possible mycotoxin contamination in various foodstuffs including legumes. PMID- 9345789 TI - Fermentation product analysis in the identification of black-pigmented bacteria from periodontitis. AB - A quantitative procedure is described for the analysis of fermentation products of eight representative black-pigmented Gram-negative anaerobic bacterial strains (Porphyromonas gingivalis 381, Porphyromonas gingivalis ATCC 33277, Prevotella intermedia ATCC 25611, Prevotella nigrescens ATCC 33563, Prevotella melaninogenica ATCC 25845, Prevotella denticola ATCC 33185, and Prevotella loescheii ATCC 15930) from oral sites in humans, using gas-liquid chromatography. This procedure for the identification of clinical isolates was carried out and the results were in agreement with those obtained by other chemical, biochemical and serological assays. The isolates were classified as Prevotella intermedia, Prevotella nigrescens and two serotype groups of Porphyromonas gingivalis, based on the quantity of fatty acids. PMID- 9345790 TI - Accumulation of insulin-gold particles in the oral apparatus of Tetrahymena after insulin pretreatment (imprinting). AB - The oral apparatus and body ciliature of untreated control cells bind insulin gold very rarely. From 1 day to 1 week after insulin pretreatment (hormonal imprinting) an enormous quantity of insulin-gold was bound by the oral cilia, completely filling the region, while the insulin-gold on the body ciliature is scattered. The binding was specific for insulin, since polyethylene glycol (PEG) gold was not bound at all. The results call attention to the binding and the increasing role of hormonal (insulin) imprinting, and particularly to the marked role of the oral region in this binding. The roles of mucocyst extrusion and specific binding by receptors are discussed. PMID- 9345791 TI - Gene-culture coevolution and sex ratios: II. Sex-chromosomal distorters and cultural preferences for offspring sex. AB - Cultural preferences for the sex of offspring may produce behavior, such as female infanticide, sex-selective abortion and sex-selective parental investment, which alter the sex ratio in a population. Empirical evidence suggests that some genetic sex-ratio distorters are located on the sex chromosomes. Interactions between cultural preferences and sex-linked sex-ratio distorters are examined. Criteria for the spread of cultural preferences and sex-chromosomal distorter alleles are derived analytically, and the coevolution of preferences and distorters is examined through numerical iteration. Evolutionary equilibria and trajectories of gene-culture interactions involving sex-chromosomal distorter alleles may produce severely male- or female-biased primary sex ratios and adult sex ratios in populations. Adult sex ratios, primary sex ratios, allele frequencies and the prevalence of cultural preferences in the population are sensitive to initial conditions and cultural transmission parameters. During the coevolutionary process phenoallelic association is observed in many cases and is associated with unusual dynamics. PMID- 9345792 TI - Modelling the feline leukemia virus (FeLV) in natural populations of cats (Felis catus). AB - A compartmental model was built in order to study the circulation and impact of Feline Leukemia Virus (FeLV) in populations of domestic cats. The model was tested with data from a long-term study of several feline populations. The study of stability shows that FeLV is maintained in the population with a stable equilibrium and a slight reduction of population size. Estimation of the transmission rate allows us to make a comparison with the values previously estimated in the literature. We compare the impact of mass vaccination or removal programmes in controlling FeLV infection, and conclude that vaccination is more efficient. PMID- 9345793 TI - Peter Smith talks to Mike Grace. PMID- 9345794 TI - Needlestick injuries. PMID- 9345795 TI - Prophylactic removal of impacted third molars. PMID- 9345796 TI - Non evidence-based fluoride prescribing recommendations. PMID- 9345797 TI - The pattern of splint usage in the management of two common temporomandibular disorders. Part I: The anterior repositioning splint in the treatment of disc displacement with reduction. AB - OBJECTIVE: To examine whether the anterior repositioning splint is suitable treatment for temporomandibular joint disc displacement with reduction and to determine the most appropriate pattern of usage. DESIGN: Prospective random control clinical trial. SETTING: Dental school clinic unit. SUBJECTS: Three groups of patients were treated, wearing the splint either during the day or at night or all the time. RESULTS: 69% of patients could be classed as improvers, by subjective and objective assessments, at final review (3 months after treatment with an anterior repositioning splint). 88% of patients who wore the splint for 24 hours per day improved over the 3-month period; this improvement was statistically significant when compared with the other two groups. CONCLUSIONS: An anterior repositioning splint is an appropriate method of treatment for disc displacement with reduction. Patients should wear the splint 24 hours a day. PMID- 9345798 TI - Computer controlled infusion of propofol for conscious sedation in dental treatment. AB - OBJECTIVE: To assess a drug delivery system that can rapidly achieve and maintain a constant blood concentration of Propofol (2,6 di-isopropyl phenol) which, in subanaesthetic doses, is an effective intravenous sedative for treating anxious or handicapped patients in dentistry. DESIGN: The clinical use of a computer controlled infusion system to induce and maintain conscious sedation with propofol was prospectively studied. Based on a 3-compartment pharmacokinetic model, the system calculates the initial bolus dose and infusion rates to achieve a user-selected target blood concentration. SETTING: Amsterdam Center for Special Dental Care. SUBJECTS: 89 patients attending for dental treatment. RESULTS: Treatment could be performed within 2 minutes after the onset of the infusion. The median therapeutic target blood propofol concentration was 2.5 micrograms/ml and the median recovery time was 9 minutes. Transient oversedation (38 procedures) could easily be treated by decreasing the target concentration. No adverse cardiorespiratory effects resulted from propofol sedation. Venous blood propofol concentrations were measured in 25 anxious patients. The kinetic data set used in this study underestimated the distribution and elimination of propofol in our patients. CONCLUSIONS: Computer controlled infusion of propofol can provide satisfactory and safe conscious sedation in dental patients. PMID- 9345799 TI - The quality of impressions for crowns and bridges received at commercial dental laboratories. AB - OBJECTIVE: To determine the quality of impressions for crown and bridge work made in general dental practice. DESIGN AND SETTING: All impressions for crown and bridge work which had been sent to four commercial dental laboratories in the UK were assessed by two examiners, each laboratory being visited on two occasions. MATERIALS AND METHODS: 290 cases which had been received by the laboratories on the days of the visits were assessed for a number of factors related to quality. There was no selection or rejection--all impressions received were examined. RESULTS: Flexible plastic trays were used for the majority of working impressions for crown and bridge work in general dental practice (72%), many had been re-used (> 13%), defects in the recording of the prepared teeth were common, and cross infection control was not routine. CONCLUSIONS: Quality standards for impressions for crown and bridge work in general dental practice in the UK are a cause for concern if the sample of cases seen in this study is typical. PMID- 9345800 TI - Angiotensin converting enzyme (ACE) inhibitors and their implications for the dental surgeon. AB - Angiotensin enzyme converting (ACE) inhibitors are a widely prescribed group of drugs used in the management of hypertension and heart failure. Several unwanted effects are associated with ACE inhibitors and this paper highlights those significant to the dental surgeon. Of particular concern is the problem of angioedema, which can be life threatening. Three case reports are presented that illustrate this problem and the management is discussed. PMID- 9345802 TI - Thinking the unthinkable--what next? AB - In December 1996, the BDA's NHS Policy Group ran a one day workshop called 'Thinking the unthinkable' in front of an invited audience from all sectors of the profession. The idea of the day was to do some preliminary forward thinking on professional issues and perhaps create a new awareness within the Association of what is happening in the rest of healthcare and to learn some lessons from them. PMID- 9345803 TI - [The HELLP syndrome as a form of late gestosis]. PMID- 9345801 TI - Fifty glorious years. AB - Since the second world war, dentistry has undergone momentous changes, not only in clinical and mechanical developments but also in changes in people's perceptions of what care their dentist can offer them. Dental graduates today will be surprised at the great difference between their own education and that of their 1946 counterparts. This is one dentist's record of the last 50 years in dentistry and how certain advancements have affected his working life. PMID- 9345804 TI - [Hormone replacement therapy and risk of breast carcinoma]. PMID- 9345806 TI - [Maternal mortality in the Czech Republic in 1995]. PMID- 9345807 TI - [The role of prenatal diagnosis in the decreasing incidence of congenital defects in the pediatric population of the Czech Republic from 1989 to 1995]. PMID- 9345805 TI - [Indomethacin in obstetrics: benefits and risks]. PMID- 9345808 TI - [Demographic aspects of induced abortion in the Czech Republic: analysis of the the 1990-1995 period]. PMID- 9345809 TI - Vaccine preventable diseases in the United States, 1996. PMID- 9345810 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 9345811 TI - Symptomatic and asymptomatic hemorrhagic cysts of the vocal folds [corrected]. PMID- 9345812 TI - Malfunctioning microdebrider: should it always be replaced? PMID- 9345813 TI - Stickler syndrome. PMID- 9345814 TI - Central auditory evaluation of patients with spasmodic dysphonia. AB - Spasmodic dysphonia is a focal laryngeal dystonia characterized by inappropriate contractions of the intrinsic laryngeal musculature. The prevalence of associated neurological findings has led to detailed investigation of the central nervous system. Previous research revealed latency abnormalities in patients' auditory brainstem responses. The present study further investigated central auditory findings in patients with spasmodic dysphonia, including brainstem and cortical function. Fourteen normal-hearing patients with spasmodic dysphonia were tested using the auditory brainstem response (ABR) and SCAN-A test of central auditory processing. The ABR estimated brainstem transmission time and evaluated auditory pathway integrity at a high stimulus rate. SCAN-A assessed the auditory cerebral cortex. Implications of these findings are discussed. We found no ABR abnormalities in subjects with spasmodic dysphonia. Positive SCAN-A findings were negligible. The ABR findings contradict previous reports. PMID- 9345815 TI - Progressive sensorineural hearing loss, subjective tinnitus and vertigo caused by elevated blood lipids. AB - The otologist frequently sees patients with progressive sensorineural hearing loss, subjective aural tinnitus and vertigo with no apparent cause. Elevated blood lipids may be a cause of inner ear malfunction on a biochemical basis. To establish the true incidence of this condition, all new patients (4,251) seen during an eight-year period were evaluated; of these, 2,332 patients had complaints of inner ear disease. All had a complete neurotologic examination, appropriate audiometric and vestibular studies and imaging, and blood tests including lipid phenotype studies. Hyperlipoproteinemia was found in 120 patients (5.1%). Most patients were found to be overweight and had additional coexisting conditions such as diabetes mellitus. Treatment with vasodilators and a 500 calorie, high-protein, low-carbohydrate diet yielded improvement of symptoms in 83% of patients within five months of initiation of treatment. PMID- 9345816 TI - Unusual localization of Castleman's disease: report of the first case in the nasopharynx. AB - We describe the first case of isolated nasopharyngeal Castleman's disease mimicking juvenile angiofibroma. Castleman's disease may appear as a local or generalized tumor-like condition, usually in the chest or abdomen, and may involve both the lymph nodes and non-nodal tissues. Since the nasopharyngeal roof is the residence of the pharyngeal tonsils (adenoids) which are rich in lymphoid tissues, such an appearance is predictable. It is emphasized that careful interpretation of radiographs may help to distinguish Castleman's disease from other tumor conditions, such as lymphoma, neurogenic tumor, or even angiofibroma, etc. But the exact diagnosis must be made on the basis of histologic confirmation. In addition to histologic classifications, clinical distinction between the localized and multicentric forms is important in selecting appropriate management. Surgical excision is the first choice and is curative in cases of localized Castleman's disease, but provides little benefit for cases of the multicentric form because of systemic manifestations and rapid deterioration. Thus, antineoplastic agents and steroids may offer an alternative form of therapy for patients with the multicentric form. PMID- 9345817 TI - Rhinophyma: treatment with CO2 laser. AB - Rhinophyma is an acne rosacea which primarily affects the midface of elderly men, and causes disfigurement as well as obstruction. There are numerous ways of treating this condition and, in our institution, a CO2 laser is the treatment of choice. PMID- 9345818 TI - The role of acoustic rhinometry in nasal provocation testing. AB - Geometric changes of the nasal airway in response to allergen challenge were measured by acoustic rhinometry (AR) and the sensitivity of the method was compared with that of rhinomanometry. Ten asymptomatic patients who suffered from ragweed allergic rhinitis were challenged out of season. The use of a custom-made noninvasive nasal adapter was an important feature of the measurement technique. A dose-dependent decrease in nasal cross-sectional area was found at and posterior to the entrance to the nasal valve. Both rhinometric and rhinomanometric methods were equivalent in sensing the changes in nasal patency due to allergen exposure (p = 0.73). Acoustic rhinometry, however, was simpler, more quickly performed and more comfortable for the subjects than was rhinomanometry by body plethysmography. AR is an alternative objective method for measurement of nasal mucosal responses, as in allergen challenge. PMID- 9345819 TI - Medical liability. PMID- 9345820 TI - Medical Injury Compensation Reform Act: Good or Bad? PMID- 9345821 TI - Physical activity and energy balance--modifiable lifestyle factors for breast cancer? PMID- 9345822 TI - Cancer in Ireland, 1994. PMID- 9345823 TI - Epilepsy and pregnancy. PMID- 9345824 TI - Urinary tract infection and contraceptive method. PMID- 9345825 TI - New developments in the treatment of psoriasis. PMID- 9345826 TI - Defining clinical signs. PMID- 9345827 TI - The behavioural phenotypes: implications for research and clinical practice. PMID- 9345828 TI - Laparoscopic management of tubal ectopic pregnancies--the Irish experience. AB - Ectopic pregnancies are being dealt with increasingly via the laparoscope on account of several advantages over conventional surgery. We commenced a Laparoscopic Ectopic service at the National Maternity Hospital in November 1995. This paper reviews the first 8 months of the service and is the first series of its kind in Ireland. All confirmed Ectopic pregnancies (EPs) between November 1995 and June 1996 are included in the study. Demographic details, diagnosis, operative details, analgesic requirements and outcome are reviewed. There were 56 tubal EPs giving an incidence of 1 per 81 live births. 35 were managed laparoscopically and were noted to have lower morbidity, analgesic requirement, reduced convalescence, shorter inpatient stay and thus lower cost. We have shown that EPs can be managed successfully via minimal access surgery in an Irish setting. In fact 22 of the last 24 cases of EPs were managed via the laparoscope. PMID- 9345829 TI - Stress in medical students. AB - The object of this study was to examine the main stressors experienced by students in an Irish medical school and their effects on the attitude of the students towards their training. It also determined the students' knowledge of how they could receive help and their attitudes towards seeking such help. Data was collected by an anonymous self-report questionnaire distributed to a fifth year medical school class in the Hilary Term of the academic year. These 63 students had chosen medicine as a career mainly because of vocational and academic factors and almost two thirds of them had always wanted to do medicine. However four were no longer happy with that choice. Fifty-four percent of them had felt like making a complaint on at least one occasion, but did not do so. The perceived problems were mainly verbal in nature. 41% said that this had affected their attendance. The main source of perceived mistreatment was consultant staff. Rates of perceived racial and sexual discrimination were low. Other stressors included examinations, financial issues and family issues. 52% of students did not know the process by which they could make a complaint and 30% felt that seeking help or advice from staff would be damaging to their future career. This study analyses these issues and suggests ways of addressing them. PMID- 9345830 TI - Prevention of ovarian cancer: a survey of the practice of prophylactic oophorectomy by consultant gynaecologists in Ireland. AB - The aim of this study was to investigate attitudes to prophylactic oophorectomy among practicing consultant gynaecologists in Ireland. An anonymous questionnaire was sent to 90 practicing consultants. A total of 68 replies were received (76%). Of these, the number who said they would remove apparently normal ovaries at the time of abdominal hysterectomy from premenopausal women in age groups < 35, 35 39, 40-44, 45-49 and > 49 years was 0 (0%), 0 (0%), 4 (6%), 29 (43%), and 46 (68%) respectively; and from postmenopausal women 60 (88%). Only 2 (3%) routinely considered oophorectomy when performing a vaginal hysterectomy. The majority of respondents said that (i) they would prescribe hormone replacement therapy in premenopausal oophorectomised women (98.5%); (ii) they did not consider unilateral oophorectomy to have a role in the prevention of ovarian cancer (84%); and (iii) they routinely discussed the question of prophylactic oophorectomy with their patients preoperatively (82%). Only 19 (27%) believed that the established figure of 10-15% of ovarian cancers could be prevented by oophorectomy at the time of hysterectomy for benign disease. 43 (63%) would perform prophylactic oophorectomy as a primary surgical procedure in women with a strong family history of ovarian carcinoma. PMID- 9345831 TI - Cervical exploration for primary hyperparathyroidism--A 25 year experience. AB - Parathyroidectomy should be considered in every patient with hypercalcaemia and primary hyperparathyroidism even if the symptoms are vague. Cervical exploration is a safe operation with very satisfactory results. Our experience in 214 patients over 25 years shows permanent postoperative normocalcaemia in 95% of cases with a complication rate of 2.8%. All patients with primary HPT, regardless of age or the severity of symptoms should be candidates for cervical exploration. PMID- 9345832 TI - Parasuicide and general practice: a pilot study. AB - General Practitioners from Cork City and its environs were sent a questionnaire regarding their experience of parasuicide in the previous twelve months. Replies were received from 133 of the 185 GPs. 189 individuals, accounting for 212 episodes of parasuicide, were seen by 78 doctors, indicating a lower level of repetition than that found in hospital-referred cases. Almost a third of doctors saw no cases, just over one fifth saw one episode and the same proportion dealt with two. A small number of general practitioners saw many cases. Regarding management, 128 (60%) were referred to Casualty, 31 of whom were also referred for psychiatric care. Thirty percent were referred directly for psychiatric care. While only fourteen were retained within general practice without referral, 40% of the GPs felt that, ideally, acts of parasuicide should be retained with more specialised advice being obtained. Furthermore, 88.1% believed that management of parasuicide should form part of an integral part of post-graduate or continued general practitioner medical training. Clearly, GPs are willing to play a more active role in the management of parasuicide. PMID- 9345833 TI - Audit of an anticoagulant clinic: doctor and patient knowledge. AB - Oral anti-coagulation with warfarin is increasingly required in the prophylaxis and treatment of vascular thrombosis and embolism. Unless the degree of anti coagulation is maintained in the narrow therapeutic range either serious bleeding or failure to prevent thromboembolism may occur. Complications may occur in up to 31% of patients. We randomly sampled 50 patients attending an anticoagulant clinic and interviewed them. We found the PTR between 2.0-4.0 in 70% patients. Their records indicated that they attended 0.9 +/- 0.5 times per month, but the patients themselves said that they had 2.4 +/- 1.7 visits per month, lasting on average 1.9 +/- 0.7 hours per visit. The mean duration of therapy was 4.3 +/- 5.4 years [range 1 month to 26 years]. Many patients perceived that they had received no education about warfarin (23%) while the majority 67% of the remainder said their doctor had educated them. Concomitant aspirin was avoided by 74% patients but 14% considered it safe in combination with warfarin; 49% patients believed that alcohol was safe in combination with warfarin. When asked about the colours and strengths of warfarin tablets, 37% of our sample were completely correct, 9% were completely incorrect and 54% were partly correct. In 16% patients they could not describe their current therapy. As doctors may adjust warfarin dosage for patients in terms of tablet colour, we asked a sample of junior doctors about the colours or strengths of warfarin tablets: 10% were completely correct, one doctor knew none of the colours or strengths and the remainder had a partial knowledge. These studies suggest that the majority of patients on warfarin are cautious about therapy and are safe in their practices. However, we feel that a significant minority may be at risk from complications because of inadequate knowledge. We suggest that improving patient understanding by education may reduce complications and lead to more stable control of anticoagulant therapy. PMID- 9345834 TI - Endometrial stromal sarcoma of the heart. PMID- 9345835 TI - Henoch-Schonlein purpura associated with adenocarcinoma [correction of adenoma] of the stomach. PMID- 9345836 TI - Appropriate use of cardiac resources needed. PMID- 9345838 TI - A new task for the information literacy training by the university libraries: electronic media and computer networking. AB - Education and training of traditional information literacy, for librarians as well as for teachers and students, is no more adequate for current demands, even if well established and not over-sophisticated. The present electronic era in the information media and technology has not been fully adopted by all its users, although all of them acknowledge its high efficiency for learning, teaching, and research. Since the year 1993/94 the members of the Institute of Scientific Information (ISI) qualified in librarianship and medicine, prepared a course for pregraduate medical students (group A) and another one for postgraduates in biomedicine (group B). Since the year 1995/96 a course has been established for bachelor studies (C). Special courses or demonstrations corresponding to the demands of those users have been organized for teachers (group D) and for members of the ISI (group E). Group A (30 students/yr, a 4 hours course in "Medical Informatics") received theoretical and practical training in principles of bibliography oriented to searching in basic biomedical databases. Group B (60-80 st/yr, 24-28 hrs/yr in "The Craft of Science") was trained in principles of scientific communication and research data presentation incl. intensive workshops with databases. Group C (50 st/yr, 30 hrs/yr in "Scientific Information") received similar training as group B but with a more practical accentation. Group D (4 persons/yr, 20 hrs/yr) and E (15 p/yr, 30 hrs/yr) proposed their topics themselves. Besides, all the groups received intense training in Internet access. The lasting feed-back contact with the majority of participants of the above courses, mainly with researchers and postgraduate students, have confirmed the advantageousness of these ISI activities. PMID- 9345837 TI - Decrease in 'pure generic' prescribing. PMID- 9345839 TI - Mechanisms of biostimulating effects of therapeutic laser in vivo. Review article. AB - The beneficial biostimulating effects induced by therapeutic laser in involved tissues was checked by a number of experimental as well as clinical works. Most of them particularly pay their attention to the description of the basic disease; there is, however, considerable imprecision in the technical characterization of the laser appliances used. Only the wave length and the power of the source are typically characterized. Further principal data are, however, missing, such as the power density, energy density, etc. These communications also suggest that their authors are not always completely familiarized with the mechanism of the stimulating action low-power laser beams in tissues. Thus, the purpose of the present communication is to summarize the existing knowledge concerning this type of treatment with considering possibilities of its use in clinical practice. PMID- 9345841 TI - How do we teach anatomy? AB - Two approaches to anatomy teaching have been studied and compared. We have concentrated on the teaching time spent for all parts of anatomy and we have compared external recommendations with the teaching time actually given to the subject at our Faculty of Medicine. In addition to that, various opinions on informational and educational aims in medical anatomy teaching are presented. The comparison mentioned above can be used to select the most useful way in which anatomy may be taught, in the light of current curricular emphases and pressures, and in the context of sound educational principles. PMID- 9345840 TI - Influence of phenytoin on cortical epileptic afterdischarges. PMID- 9345842 TI - Relationship of medicine and physician toward patient at the advent of 3rd millennium. PMID- 9345843 TI - Feline poxvirus infection. A case report. AB - Poxvirus infection of a domestic cat is reported. The clinical signs consisted of skin lesions only, which healed within two and a half months. Histopathology revealed cytoplasmatic inclusion bodies typical of pox virus infection. Virus particles morphologically related to the genus orthopoxvirus were detectable in the embedded skin tissue and in skin scraping by electron microscopy. No specific lesions were observed in the chick embryo chorioallantoic membrane inoculated with an extraction from skin scabs of the cat. PMID- 9345844 TI - Human genome project Europe. PMID- 9345845 TI - Time to reassess chiral aspects of beta-adrenoceptor antagonists. PMID- 9345846 TI - Improved therapies for Parkinson's disease: life beyond dopamine D2/D3 receptor agonists. PMID- 9345847 TI - Re-classification of NSAIDs. PMID- 9345848 TI - Re-addressing relative selectivities. PMID- 9345849 TI - Changing the countenance of pharmacology courses in medical schools. PMID- 9345850 TI - Tumour necrosis factor alpha and interleukin 1 signalling: do MAPKK kinases connect it all? AB - The potent pro-inflammatory cytokines tumour necrosis factor alpha (TNF-alpha) and interleukin 1 (IL-1) are capable of triggering biologically similar effects through activation of the same set of transcription factors. Based on recent findings it is now becoming evident that certain members of the mitogen-activated protein kinase kinase (MAPKK) kinase protein family serve to integrate the individual signal transduction pathways that are initiated by the two cytokines into an array of parallel and common signalling cascades. The link between the receptor proximal, signal-specific intracellular events and the common MAPKK kinases appears to be made by a new class of proteins known as TNF receptor associated factors (TRAFs). Here, Jorg Eder describes how TNF-alpha and IL-1 use different, pathway-specific TRAFs to activate the same MAPKK kinase-controlled cascades. PMID- 9345851 TI - A protein-mediated mechanism for the DNA sequence-specific action of topoisomerase II poisons. AB - Chemical agents able to interfere with DNA topoisomerases are widespread in nature, and some of them have outstanding therapeutic efficacy in human cancer and infectious diseases. DNA topoisomerases are essential enzymes that govern DNA topology during fundamental nuclear metabolic processes. Topoisomerase interfering compounds can be divided into two general categories based on the mechanism of drug action: poisons and catalytic inhibitors. In past years, investigations of the DNA sequence selectivity of topoisomerase II poisons have identified structural and molecular determinants of drug activity, and indicated that the drug receptor is likely to be at the protein-DNA interface. Moreover, the available results indicate that the biologically relevant DNA-binding activity of topoisomerase poisons is basically protein-mediated and this is discussed in this issue by Giovanni Capranico and colleagues. This suggests that topoisomerase poisons may represent a useful paradigm for small compounds able to bind to protein-DNA interfaces in a site-selective manner, thus increasing the affinity of DNA-binding proteins for specific genomic sites. PMID- 9345852 TI - Clinical and molecular aspects of motoneurone diseases: animal models, neurotrophic factors and Bcl-2 oncoprotein. AB - Animal models of motor neurone disease (MND) are being increasingly used for screening molecules with clinical potential. A number of different treatments to decrease the progression of neuronal cell loss have been proposed; these include: Bcl-2 (B-cell leukaemia oncogene-2), neurotrophic factors, glutamate receptor inhibitors and Ca2+ channel antagonists. In this review Yves Sagot, Richard Vejsada and Ann C. Kato focus on the effects of neurotrophic factors and Bcl-2, both of which have been shown to prevent cell death in various experimental paradigms. Studies performed in animal models of MND have confirmed the potential of these molecules to support motoneurone survival. Some of them have been shown to act in synergy and these results are discussed in the context of molecular mechanisms leading to collaborative and synergistic activities, and also with respect to presumptive subpopulations of motoneurones, which express diverse receptors for neurotrophic factors. Finally, the current status of clinical trials for amyotrophic lateral sclerosis using neurotrophic factors will be discussed, as well as recent reports that neurotrophic factors can exert adverse effects on neuronal survival. PMID- 9345854 TI - [Spirapril in the therapy of hypertension] [In Process Citation] PMID- 9345853 TI - Adenosine A2A receptor antagonists as new agents for the treatment of Parkinson's disease. AB - There is now good reason to believe that blockade of the adenosine A2A receptor could be of value in the treatment of Parkinson's disease. Peter J. Richardson, Hiroshi Kase and Peter G. Jenner review the actions of this receptor in the striatum, emphasizing its ability to modulate the neuronal activity of striatal GABA-releasing output neurones, and showing that recently developed A2A receptor antagonists are capable of reducing the disabling effects of nigral cell degeneration in primates. They conclude that such antagonists may be useful as novel therapeutic agents for the treatment of Parkinson's disease. PMID- 9345895 TI - Multiple human endogenous retrovirus (HERV-K) loci with gag open reading frames in the human genome. AB - Human endogenous retrovirus HERV-K is present in 25-30 copies per haploid human genome. At least one of these loci is capable of producing full-length Gag protein, high amounts of which have been detected in germ cell tumors and derived cell lines. The latter display HERV-K Gag-encoded retroviral particles. Here, we employed the protein truncation test (PTT) in combination with a monochromosomal hybrid mapping panel to identify the human chromosomes harboring HERV-K gag genes with an open reading frame for Gag protein. Eight human chromosomes were found to contain intact HERV-K gag genes. PTT results were corroborated by partial sequencing of subregions from different HERV-K gag genes. The high number of HERV K Gag open reading frames supports the idea of retroviral sequences retaining a biological benefit in the human genome. PMID- 9345896 TI - A high resolution GBG-banded karyotype of the Atlantic bottlenose dolphin, Tursiops truncatus: generation of an ideogram, and NOR localization by fluorescence in situ hybridization. AB - A high resolution karyotype was prepared using GBG-banded chromosomes from kidney epithelial cell lines of the Atlantic bottlenose dolphin, Tursiops truncatus. The karyotype was used to generate a banded ideogram of T. truncatus that will facilitate cytogenetic analyses essential for comparison of dolphin chromosomes with those of potentially related terrestrial vertebrates. Fluorescence in situ hybridization analysis of the karytype using a rodent (Geomys bursarius) 28S ribosomal DNA (rDNA) probe allowed localization of four nucleolar organizer (NOR) regions to the short arms of two chromosome pairs. FISH mapping, using DNA probes, is dependent on a pre-existing banded ideogram, and provides the means to initiate physical mapping of the dolphin genome. PMID- 9345897 TI - Complex FISH probes for the subtelomeric regions of all human chromosomes: comparative hybridization of CEPH YACs to chromosomes of the Old World monkey Presbytis cristata and great apes. AB - We have generated a human subtelomere probe panel, utilizing well characterized CEPH YACs, for the investigation of human chromosome pathology and evolution through fluorescent in situ hybridization (FISH). Region-specific FISH probes will be extremely valuable for detecting cytogenetically cryptic telomere abnormalities. Here, we present the first comparative mapping study (with 29 subtelomere probes and 6 chromosome paints) to the Old World monkey Presbytis cristata, followed by hybridizations to the great apes, gorilla and orangutan, when rearrangements were detected. We observed that the position of telomere associated genomic sequences has been only moderately conserved during primate evolution. YAC 364f9, specific for the subtelomeric long arm of human chromosome 3, contains an evolutionary inversion breakpoint that was involved in independent chromosome rearrangements in P. cristata and gorilla. PMID- 9345898 TI - EST00444 (D13S308E) maps to 13q12.1. PMID- 9345899 TI - In situ hybridisation mapping of genomic clones for five human respiratory chain complex I genes. AB - Using human and bovine short cDNA sequences as probes we screened human cosmid and P1 libraries for components of the complex I multi-subunit enzyme of oxidative phosphorylation. We isolated genomic recombinants encoding cI-B8 (gene NDUFA2), cI-B14 (gene NDUFA6), cI-B14.5a (gene NDUFA7), cI-ASHI (gene NDUFB8) and cI-23kD (gene NDUFS8). Genomic versions of these genes have not been previously cloned in the human although they are represented as anonymous entries in public cDNA databases. By using the derived genomic clones for in situ hybridisation studies we determined the following chromosome locations: NDUFA2, 5q31; NDUFA6, 21q22; NDUFA7, 20p13; NDUFB8, 12q21; NDUFS8, 3q28. PMID- 9345900 TI - Assignment of the gene for very-long-chain acyl-CoA dehydrogenase (Acadvl) to mouse chromosome band 11B2-B5 by in situ hybridization. PMID- 9345901 TI - Sex chromosome aneuploidy in sperm-derived pronuclei, motile sperm and unselected sperm, scored by three-color FISH. PMID- 9345902 TI - Molecular cloning and mapping of a human cDNA (PA2G4) that encodes a protein highly homologous to the mouse cell cycle protein p38-2G4. AB - We have identified a novel human gene with strong homology to the mouse Pa2g4 cell cycle gene. This novel gene (called PA2G4) belongs to a gene family with members in several chromosome regions: 3q24-q25, 6q22, 9q21, 12q13, 18q12, 20p12 and Xq25. A composite cDNA of 1697 nucleotides was isolated. The sequence of this cDNA predicts a protein of 394 amino acids. The deduced amino acid sequence of this human protein shows very strong homology to the mouse protein p38-2G4. The cDNA analyzed probably corresponds to a functional copy found at 12q13. PMID- 9345903 TI - Genome composition in Venezuelan spiny-rats of the genus Proechimys(Rodentia, Echimyidae). I. Genome size, C-heterochromatin and repetitive DNAs in situ hybridization patterns. AB - The genome sizes of the Venezuelan spiny-rats Proechimys guairae guairae (2n = 48) and P. trinitatis (2n = 62) were evaluated and proved to be 12.5 +/- 0.5 pg and 12.6 +/- 0.3 pg respectively, the highest so far recorded among mammals; also the C-heterochromatin (32.7%, Coefficient of Variation [CV] 3.8 and 35.8%, CV 4.4) and GC (44.2%, CV 2.7 and 43.6%, CV 2.9) contents are very high. Highly repetitive (rep) DNAs were isolated from restriction enzyme digested genomic DNAs of both species. The intra- and inter-specific chromosomal allocations of these rep DNAs were analyzed by direct and cross-hybridizations. Results show that the two genomes harbour several rep DNA families which show both species-specificity and interspecific relatedness in their in situ hybridization patterns. The rep DNA families show an equilocal distribution at both the pericentromeric areas of all chromosomes and in the whole arms of two pairs of the uniarmed group, suggesting co-evolution of the rep DNAs. P. g. guairae BamHI digested DNA, when cloned and sequenced, proved to consist of a long "composite" unit (1,239 bp) containing two copies of each of 75-bp and 110-bp internal subrepeats. Karyotype restructuring between P. g. guairae and P. trinitatis, mainly due to Robertsonian changes, was accompanied by slight intra- and intergenomic movements of the putative satellite DNA families within stable genome sizes and C-heterochromatin contents. We discuss the findings obtained in Proechimys in the light of those regarding the kangaroo rat, the pocket gopher and the house mouse; they support the idea that karyotype restructuring could be the expression of molecular driven events of rep DNA amplification and homogenisation through non-homologous chromosomes. PMID- 9345905 TI - Assignment of ACTBP8 (alias ACTBP2) within or close to human chromosome band 6q13 using a radiation hybrid panel. PMID- 9345904 TI - PCP4 maps between D21S345 and P31P10SP6 on chromosome 21q22.2-->q22.3. PMID- 9345906 TI - Assignment of FGF12, the human FGF homologous factor 1 gene, to chromosome 3q29- >3qter by fluorescence in situ hybridization. PMID- 9345907 TI - M33, a mammalian homologue of Drosophila Polycomb localises to euchromatin within interphase nuclei but is enriched within the centromeric heterochromatin of metaphase chromosomes. PMID- 9345908 TI - Assignment of fragile site 8E (FRA8E) to human chromosome band 8q24.11 adjacent to the hereditary multiple exostoses 1 gene and two overlapping Langer-Giedion syndrome deletion endpoints. PMID- 9345909 TI - Genomic structure of the human PLCD1 (phospholipase C delta 1) locus on 3p22- >p21.3. AB - A large-scale sequencing analysis of genomic DNA in the vicinity of homozygous deletion on chromosome 3p found in a lung cancer cell line disclosed that the gene encoding phospholipase C delta 1 (PLCD1) is located just distal to the region removed by the deletion We report the genomic structure of this gene, which consists of 15 exons and spans about 22 kb, and its precise localization to chromosome 3p22-->p21.3. PMID- 9345910 TI - Assignment of p53 binding protein (TP53BP2) to human chromosome band 1q42.1 by in situ hybridization. PMID- 9345911 TI - Assignment of mitotic arrest deficient protein 2 (MAD2L1) to human chromosome band 5q23.3 by in situ hybridization. PMID- 9345912 TI - Experimental assessment of the detection limit of genomic amplification by comparative genomic hybridization CGH. AB - Artificial amplicons of known size, constructed by use of YACs featuring human 8p12 and 12q13, were analyzed by comparative genomic hybridization (CGH). A minimum of 15 Mb of overrepresented DNA sequences could be detected. The sensitivity is (1) not dependent on the chromosome site and (2) related to the size of the amplicon, decreasing with decreasing size. PMID- 9345913 TI - Fertility investigations in the F1 hybrid and backcross progeny of cattle (Bos taurus) and yak (B. grunniens) in Mongolia. AB - Investigations conducted in Mongolia into the sterility of the male khainag, an F1 hybrid animal resulting from crossing cattle (Bos taurus, 2n = 60) with yaks (B. grunniens, 2n = 60), are reported. Reduced numbers of spermatogonia appear to characterise the testicular tubules of the khainag, and despite the identical cytological appearance of the two parental karyotypes, synaptic anomalies are seen at meiotic prophase in primary spermatocytes. The female khainag is fertile and can be backcrossed to cattle or yak bulls to produce a B1 backcross animal, the ortoom. Further backcrossing of ortoom females to cattle or yaks will yield a B2 backcross animal, the usanguzee. The impression is gained of better meiotic pairing in the backcross animals than in the khainag. The "Haldane Rule" is followed perfectly by the cattle x yak hybrid; namely, sterility is confined to the male. PMID- 9345914 TI - Mapping the distribution of the telomeric sequence (T2AG3)n in rock-wallabies, Petrogale (Marsupialia: Macropodidae), by fluorescence in situ hybridization. I. The penicillata complex. AB - The eight Petrogale (rock-wallaby) species of the penicillata complex have a variable rate of karyotypic evolution, with species differing from the ancestral karyotype by two to six rearrangements. The distribution of the predominant vertebrate telomeric sequence (T2AG3)n was examined by fluorescence in situ hybridization (FISH) to determine if this sequence is retained during centric fusion events or is involved in other rearrangements. In all submetacentric chromosomes derived by centric fusions, the telomeric sequence was identified at or near the centromere, indicating that the (T2AG3)n sequence is consistently retained. In two acrocentric chromosomes, derived by centromeric transpositions from submetacentric fusion chromosomes, an interstitial signal was observed at the presumed site of the fusion. This represents the identification of a novel mechanism by which the (T2AG3)n sequence may become interstitial. Other interstitial telomeric signals were identified just below the centromere of chromosome 1 and interstitially on chromosome 4 in all eight species of the penicillata complex. These may be related to, respectively, the formation of euchromatic short arms on chromosome 1 and a more ancient rearrangement of chromosome 4 within marsupials. PMID- 9345980 TI - International Society for Child and Adolescent Injury Prevention. PMID- 9345981 TI - The history of childhood accident and injury prevention in England: background to the foundation of the Child Accident Prevention Trust. PMID- 9345982 TI - America's experiment with motor vehicle safety regulation. PMID- 9345983 TI - Injury prevention: an uphill battle. PMID- 9345984 TI - What's in a name? Comments on the use of the terms 'accident' and 'injury'. PMID- 9345985 TI - Accident prevention--injury control--injury prevention--or whatever? PMID- 9345986 TI - Sensory deficit and the risk of pedestrian injury. AB - OBJECTIVES: To examine the association between sensory deficit and the risk of child pedestrian-motor vehicle collisions. SETTING: The Auckland region of New Zealand. METHODS: A community based case-control study was conducted. Cases (n = 190) were all children (< 15 years) killed or hospitalised as a result of a pedestrian injury occurring on a public road between 1 January 1992 and 1 March 1994. Controls (n = 479) were a random sample of the child population. RESULTS: The risk of pedestrian injury for children whose parents reported abnormal vision was over four times that of children with reported normal vision (odds ratio = 4.25, 95% confidence interval 1.68 to 10.8). The risk of injury for children whose parents reported abnormal hearing was close to twice that of children with reported normal hearing (odds ratio = 1.73, 95% confidence interval 0.83 to 3.61). CONCLUSIONS: Children with sensory deficits constitute a high risk group for pedestrian injuries. Paediatricians caring for children with sensory impairments should be aware of this increased risk. PMID- 9345987 TI - Gender differences in injuries among rural youth. AB - GOAL: This paper presents injury data from the first year of a three year longitudinal study of risk taking behaviors among adolescents. SAMPLE: Study subjects were a cohort of 758 rural students from Maryland's Eastern Shore who were in the eighth grade in 1987. METHODS: Students completed a 45 minute, self administered survey in which they reported numbers of injuries experienced in the past year, risk taking behaviors, anger expression, delinquency, alcohol and drug use, physical exercise, work experience, and level of parental supervision. In addition, students had their height and weight measurements taken by trained research staff and completed a self rating of pubertal development using Tanner drawings. RESULTS: Slightly more than half (53.2%) of the boys and over one third (37.7%) of the girls reported experiencing one or more medically attended injuries during the last year. Poisson regression analyses were conducted to estimate the extent to which gender differences in injuries could be accounted for by adolescent behaviors. Gender effects became non-significant when adjustments were made for risk taking, school discipline problems, and exercise frequency. Gender differences in injuries were reduced but remained significant when substance use, employment, and anger were controlled. Poisson regression analyses were conducted separately for males and females to assess whether factors associated with injuries were similar across genders. For boys, risk taking, anger, and school discipline problems were significantly related to number of injuries. Boys with a low body mass index and late pubertal development (mean ratio 3.09), as well as those with high body mass index and early pubertal development (mean ratio 2.16), reported greater numbers of injuries than average boys. For girls, substance use, cruising, risk taking, anger, and exercise frequency were significantly associated with injuries. Girls with an early onset of menses reported, on average, twice the number of injuries than those who were on time. Girls with high body mass index who were late in their pubertal development reported, on average, five times more injuries than other girls. CONCLUSIONS: Although gender is a significant risk factor for injuries, certain behaviors like risk taking, school related delinquency, and physical exercise partially explain the higher number of injuries among adolescent males in this study. For both males and females, indicators of pubertal and physical development are important factors to consider in studies of injuries during early adolescence. PMID- 9345988 TI - Head injuries in helmeted child bicyclists. AB - OBJECTIVE: To determine the characteristics and the severity of head and facial injuries to helmeted child bicyclists, and whether the helmet contributed to the injury, and to study factors related to bicycle accidents. DESIGN: Retrospective review of two case series. Children sustaining head injury while not wearing helmets were studied as a form of reference group. SETTING: Large paediatric teaching hospital. SUBJECTS: 34 helmeted child bicyclists and 155 non-helmeted bicyclists, aged 5-14 years. MAIN OUTCOME MEASURES: Number of injuries, type of injuries, injury severity score, deaths, and accident circumstances. RESULTS: 79% of the head injuries of the helmeted child group were mild and two thirds of these had facial injuries. Children in the helmet group were in a greater proportion of bike-car collisions than the no helmet group and at least 15% of the helmets were lost on impact. There were no injuries secondary to the helmet. CONCLUSIONS: Most of the head injuries sustained by the helmeted children were of mild severity and there was no evidence to suggest that the helmet contributed to injury. Nevertheless, consideration should be given to designing a facial protector for the bicycle helmet and to improvement of the fastening device. PMID- 9345989 TI - Patterns of injury in children and adolescents presenting to a South African township health centre. AB - OBJECTIVES: To describe the patterns and causes of childhood injury presenting to a South African township health centre in 1991. DESIGN: Retrospective review of clinic held case notes. SETTING: Typical South-African urban township within Greater Johannesburg. SUBJECTS: 695 subjects aged 0-19 years presenting as a direct result of injury. RESULTS: Overall rates of presentation for injury were 6297/100,000/year (95% confidence interval 5463 to 7131); 35% of injuries were caused by violence, 14% by traffic, and 51% by other unintentional causes (such as falls and sport injuries). Males had higher rates of presentation than females for violent (p < 0.001) and unintentional injuries (p < 0.01), but rates were similar for traffic injuries. The highest rates were for injuries caused by violence in 15-19 year-old males and were 9319/100,000/year. CONCLUSIONS: Rates are lower than in more developed countries. However, they appear to represent the more severe end of the spectrum of injury severity. The rates are similar for those below age 10 years and higher for those above age 10 years compared with severe injury rates in other studies. These data are likely to underestimate true rates. The risk of injuries caused by violence increase with age and these injuries are more serious than those due to other causes. Males are at higher risk for all types of injury except traffic injury. PMID- 9345990 TI - Determinants of modern health care use by families after a childhood burn in Ghana. AB - OBJECTIVES: This study examined determinants of modern health care use by families after their child aged 0-5 years sustained a burn injury in the Ashanti Region of Ghana. METHODS: A community based survey of children aged 0-5 years was conducted in 50 enumeration areas in the region. Mothers of all children with scars as evidence of a burn were selected for a follow up interview using a standard questionnaire two to three months later. Determinants of health care use were investigated through a multivariate logistic regression using interview responses from mothers of 617 children for whom report on some treatment was given. RESULTS: Overall, 48% of the burned children were taken to a modern health facility for treatment. Of those taken to a modern health facility, 68% were sent within 24 hours of the burn event. Factors with large adjusted odds ratios for modern health care use included wound infection, burns covering 6% or more of the body surface, and third degree burns. Compared with scalds, children with contact and flame burns were less likely to be taken to a health facility, as were burns to rural children, and those given first aid treatment at home. CONCLUSIONS: It is concluded that families, particularly rural residents, should be educated about appropriate health care seeking practices after a burn. PMID- 9345991 TI - Injury prevention program in primary care: process evaluation and surveillance. AB - OBJECTIVES: To carry out process evaluation and surveillance in a community oriented primary care program for injury prevention among children 0-2 years old (n = 306). SETTING: Mother and child health clinic in a defined area of Western Jerusalem. METHODS: An injury prevention program was integrated into the routines of the mother and child health clinic. The program consisted of injury surveillance and counselling using a developmental approach, regarding car safety and the prevention of falls, burns, suffocation, poisonings, cuts, drowning, and electrocution. Process evaluation and surveillance were based on records integrated into the child's personal file in the clinic. RESULTS: Process evaluation indicated that counselling coverage was 73% in the 0-5 month age group and decreased to 48% in the second year of life. The mean number of topics discussed with the parents was 6.6 (out of nine) for the 0-5 months age group, 13.6 (out of 18) for the 6-11 month group, and 15.7 (out of 18) for those 1-2 years old. Injury surveillance activities were complete for 66% of the children, incomplete in 32%, and not done in 2%. CONCLUSIONS: The results indicate that it is feasible to integrate an injury prevention program into primary care, and that process evaluation is important in detecting problems and improving performance of the program's activities. PMID- 9345992 TI - Energy damage and the 10 countermeasure strategies. 1973. PMID- 9345993 TI - Methodologic issues in injury case-control studies. AB - In this paper some methodological problems particularly relevant to case-control studies of injury are illustrated by reference to previous childhood injury case control studies. In contrast to studies of disease, where 'person time' constitutes the observational experience of interest, in injury studies person time engaged in a particular activity to often more appropriate. The implications for the definition of the study base are discussed. The potential for hospital admission bias in injury case-control studies is considered along with potential strategies for avoiding it. The importance of errors in exposure measurement, including those arising from inappropriate induction time assumptions, are illustrated. Finally, the potential for bias resulting from the combination of etiologically unrelated injury outcomes into a single outcome measure is illustrated and discussed. PMID- 9345994 TI - The Child Accident Prevention Trust (UK). PMID- 9345996 TI - 'Interdisciplinary' and 'multidisciplinary' are not synonymous. PMID- 9345995 TI - The role of health education in childhood injury prevention. PMID- 9345997 TI - Sweden's pioneering child accident programme: 40 years later. PMID- 9345998 TI - Deregulation and product safety. PMID- 9345999 TI - Regulating risk to children. PMID- 9346000 TI - Non-intentional asphyxiation deaths due to upper airway interference in children 0 to 14 years. AB - OBJECTIVE: This study was undertaken to identify avoidable, contributing factors associated with non-intentional asphyxiation deaths due to upper airway interference in children 0 to 14 years. DESIGN: Historical population based incidence study. METHODS: All postneonatal and childhood deaths by asphyxiation from 1985 to 1994, using appropriate ICD-9-CM codes, were compiled from the Victorian government legislated paediatric mortality surveillance system. Recent cases were identified from the State Coroner's Office. Case definition included children under 15 years who died from upper airway interference such as facial occlusion, head and neck entrapment, rope or cord strangulation, or foreign body. RESULTS: Of the identified 42 deaths, eight (19%) were caused by a foreign body in the airway, five (12%) were due to facial occlusion, 16 (38%) were due to ropes and similar material (seven were homemade rope swings), and 13 (31%) were caused by entrapment (seven were in cots or beds). The average annual rate for asphyxiation deaths by all causes for children 0 to 14 years was 4.7 million. Infants under 1 year had a rate of 20.1/million, while the rate for 10 to 14 year olds dropped to 2.0/million. CONCLUSION: Rope swings and rope material are inherently dangerous and frequently prove fatal, especially for older children. For infants, environmental factors are important; in particular food and bedding. Prevention strategies need to be developed that include obligatory standards for the design and manufacture of products for children, appropriate labelling and warnings, and education for children, their carers, and health care professionals. PMID- 9346001 TI - Evaluation of the Think First head and spinal cord injury prevention program. AB - OBJECTIVE: Evaluation of the impact of the Think First head and spinal cord injury prevention program on knowledge, attitudes, and behavior of 11-15 year old students toward injury risks and preventive strategies. SETTING: Three junior high and three senior high schools in rural and urban areas of Washington state. METHODS: Questionnaire survey before intervention, two weeks and three months after intervention to assess knowledge, attitude, and self reported behavior change. Observations of students as they left school property to determine bicycle helmet and seat belt use. RESULTS: Little impact on attitudes and no consistent change in knowledge or self reported behaviors. Too few students rode bicycles to accurately assess helmet use; no consistent change in seat belt use. CONCLUSION: The Think First program appears to have little impact on changes in knowledge, self reported behavior, or observed behavior. Other strategies to decrease injuries in adolescents may be more successful. PMID- 9346002 TI - Children and bicycles: what is really happening? Studies of fatal and non-fatal bicycle injury. AB - OBJECTIVES: The objectives of the study were to ascertain the causes of accidents, injuries, and deaths in children who ride bicycles. Fatality and injury rates were also studied in order to compare with other studies. METHODS: Two studies of children were undertaken in children aged less than 15 years. In the first (retrospective fatality study), children who died as a result of a bicycle incident during the period 1981-92 were reviewed. In the second (prospective injury study) data were obtained prospectively between April 1991 and June 1992 about children who were injured while riding a bicycle and treated at a public hospital in Brisbane. RESULTS: Study 1: fatality rates for boys were twice those for girls. The rate was highest for boys of 14 years in the metropolitan area at 6.23/100,000. All deaths involved vehicles, and the majority involved head injury or multiple injuries including head injury. Study 2: similar numbers of children were injured at onroad and off-road locations. Faculty riding was described by the rider or caregiver as the cause in 62.5% of cases. The most common time of injury was between 3 and 6 pm on both school and non-school days. Only 5.5% of all incidents involved a moving vehicle. CONCLUSIONS: Bicycle riding by children is a common cause of injury, particularly for boys. Equal numbers of injuries occurred on the road as at other locations. Faulty riding caused most accidents. Injury prevention for bicycle riders should involve not only compulsory wearing of helmets, but should also include education and training about safe riding habits, separation of motorised vehicles from bicycles, modified helmet design to incorporate facial protection, and improved handlebar design. PMID- 9346004 TI - Fatal and hospitalized agricultural machinery injuries to children in Ontario, Canada. AB - OBJECTIVES: To assess rates and patterns of agricultural machinery injuries in farm children in order to both determine priorities and develop strategies for injury control in this population. METHODS: Coroners' files and hospital discharge data were examined for Ontario farm children aged 0-19 who had agricultural machinery injuries over a five year period ending 31 March 1990. Injury rates were described by age, sex, geographic region, type of machinery, and mechanism of injury. Common patterns of injury deserving of priority for prevention were then identified and described. RESULTS: 283 machinery injuries to children were identified. Injury rates were 116 and 25/100,000/year for boys and girls respectively. Boys were at increasing risk relative to girls as their ages increased. Young children were at greatest risk for fatal injury. There is a prominent summer peak in occurrence. The farm tractor was the machine most commonly associated with these injuries (33.2%), and entanglement, usually of clothing, was the mechanism cited most often (36.3%). The case fatality ratio (ratio of hospitalizations:deaths) was generally low whether assessed by machinery type or by mechanism of injury. This provides an indication of the lethality of these injuries. Common patterns associated with injury risk included: (1) inadequate supervision of small children; (2) permitting children to be in the area of moving or unguarded machinery; (3) allowing children to accompany workers using farm machinery; and (4) having children performing work related tasks inappropriate for their age. CONCLUSIONS: Machinery related injuries are not uncommon in farm children and have a high case fatality rate. These rates changed little over the five year study period. Feasible strategies for prevention of these injuries, four of which are presented here, need to be developed and implemented by public health professionals working in cooperation with members of the agricultural industry. PMID- 9346003 TI - Biosocial variables and auditory acuity as risk factors for non-fatal childhood injuries in Greece. AB - OBJECTIVES: To examine whether biosocial variables and auditory acuity are risk factors for injuries among children. SETTING: Children with injuries who presented at the emergency clinics of one of the two university hospitals for children in Athens, Greece between December 1993 and April 1994. METHODS: 144 children aged 5-14 years, residents of Athens, were brought to the emergency clinics for a moderate to severe injury. For each of these children one hospital control, matched for age and sex, and one classmate control similarly matched were identified. A standard interview form was completed for all 432 children and acouometric and tympanometric examinations were performed in each of them. Analysis was done through conditional logistic regression. RESULTS: The likelihood of an accident was higher in children of younger fathers (odds ratio (OR) = 0.7, p = 0.04), children of mothers with non-professional jobs (OR = 1.9, p = 0.03) as well as in children of higher birth order (OR = 1.7, p = 0.01), in those with predominantly other than parental daily supervision (OR = 2.6, p = 0.001), and those with a history of previous accident (OR = 1.3, p = 0.002). Somatometric factors, school performance, use of corrective eyeglasses and subnormal auditory acuity were not found to be risk factors, but auditory imbalance and abnormal tympanograms were positively related to the risk of childhood injury (OR = 2.6, p = 0.02; and OR = 2.3, p = 0.08 respectively). CONCLUSIONS: the findings of this study underline the importance of attentive supervision and safety training of children living in modern cities; they also suggest that children with auditory imbalance and history of an accident are at higher injury risk and they should be targeted with specific intervention programs. PMID- 9346005 TI - Young on-road motorcyclists in New Zealand: age of licensure, unlicensed riding, and motorcycle borrowing. AB - OBJECTIVES: The study aimed to determine the prevalence of unlicensed riding and motorcycle borrowing among young motorcyclists, and to document their perceptions of how they would be affected if the minimum age of licensure were raised. METHODS: Motorcycling was investigated as part of the Dunedin Multidisciplinary Health and Development Study, a broad longitudinal study of the health, development, attitudes, and behaviours of a birth cohort. Young motorcyclists, who had ridden on-road during the year before their interview at age 18 years, completed a computer administered questionnaire containing questions about licensure, riding frequency, and motorcycle borrowing. RESULTS: Of the 217 motorcyclists identified, 36% were licensed, 54% had ridden once a month or less frequently, and 72% had usually ridden a borrowed motorcycle during the one year recall period. Significantly more licensed than unlicensed riders and owners than borrowers reported higher exposure and significantly more licensed than unlicensed riders were owners. Most licensed riders (86%) had ridden on public roads before licensure, and many (54%) thought that they would have been much affected by a higher minimum age of licensure. CONCLUSIONS: More stringent enforcement of existing licensing regulations, tougher penalties for breaching graduated driver licensing restrictions, raising the minimum age for motorcycle licensure, and prohibiting the sale or lending of motorcycles to unlicensed riders are possible injury prevention strategies. PMID- 9346006 TI - Attitudes to and use of baby walkers in Dublin. AB - OBJECTIVES: To identify the rate of baby walker use, parental attitudes, and associated injuries. DESIGN: Parents of babies attending clinics for developmental assessment were surveyed by self administered questionnaire about their use, attitudes, and history of injuries associated with walkers. SETTING: Dublin, Ireland. SUBJECTS: Parents of 158 babies. RESULTS: Fifty five per cent of the sample used a walker. The main reasons for doing so included babies' enjoyment of them and the fact that the walker was used for an older sibling. Although none of the users listed safety concerns as a reason to stop using the walker, non-users (45%) did so; 12.5% of the users had at least one walker related injury. CONCLUSIONS: Parents of babies who use a walker perceive them as beneficial. However these babies are placed at unnecessary risk. It behoves all health professionals and child carers to alert parents to these dangers and the sale of walkers should be reviewed. PMID- 9346007 TI - Young children in traffic. 1970. PMID- 9346008 TI - The evaluation of community based injury prevention activity: the UK perspective. PMID- 9346009 TI - The National SAFE KIDS Campaign (USA). PMID- 9346010 TI - A review of educational and legislative strategies to promote bicycle helmets. PMID- 9346011 TI - Head injuries in helmeted child bicyclists. PMID- 9346012 TI - Children shooting guns: a failure in product design. PMID- 9346013 TI - The Quebec Sports Safety Board: a governmental agency dedicated to the prevention of sports and recreational injuries. PMID- 9346014 TI - Effectiveness of injury prevention counseling. PMID- 9346015 TI - Primary care counselling for injury prevention: where is the evidence? PMID- 9346016 TI - Who's prepared for advocacy? Another inverse law. AB - OBJECTIVES: To examine the characteristics of parents responding to a petition calling for greater efforts to ensure the safety of children as pedestrians and to contrast factors predictive of advocacy with risk factors for child pedestrian injury. SETTING: The Auckland region of New Zealand. METHODS: Parents participating in the Auckland Child Pedestrian Injury Study, a community based case-control study, were invited to support a series of recommendations based on the study results, by signing and returning a petition that was to be delivered to the New Zealand Minister for Transport. Characteristics of petitioners were determined by linking their petition responses to the study questionnaires using an unique identifier. The characteristics of petitioners and nonpetitioners were summarised using odds ratios. RESULTS: 31% of parents signed and returned the petition; 19% were parents of cases and 36% were parents of controls. The sociodemographic groups whose children were at the lowest risk of pedestrian injury were the most likely to return the petition. Children in the most disadvantaged socioeconomic group and children of Pacific Island parents were at greatest risk of injury but the parents of these children were the least likely to respond to the petition. CONCLUSIONS: The frequency with which parents advocate for child safety varies inversely with the need for it. Models of health promotion based on community ownership and empowerment alone are unlikely to address the steep socioeconomic gradients in childhood injury mortality. PMID- 9346017 TI - The contribution of physicians to childhood injury prevention in France. AB - OBJECTIVES: The objective of this study was to determine what injury control interventions are currently carried out by physicians and to examine how these interventions could be more effective. SETTING: Surveys were conducted among the three main groups of physicians who provide primary care to children in France- private practice pediatricians (PPPs), well-child clinic pediatricians (WCCPs), and general practitioners (GPs). METHOD: A representative sample of each of the three groups of physicians were interviewed by telephone, using a computer assisted telephone interview system, in December 1993 or February 1994. RESULTS: Responses demonstrated that most physicians felt they could play an important part in injury prevention but that many had inadequate knowledge of injury related mortality rates in children. Most PPPs and WCCPs usually provided counseling on safety in relation to developmental changes in children. Few physicians gave recommendations about appropriate first responses to emergencies. Printed material, designed for parent education, was provided by many PPPs and WCCPs, but was usually absent from the offices of GPs. Participation in group education sessions was common among WCCPs but rare among PPPs and GPs. Many physicians expressed skepticism regarding the efficacy of their interventions in injury control. CONCLUSION: A number of recommendations are made to those in government agencies or elsewhere who could help physicians to improve childhood injury prevention, for instance by regular publication of data on childhood injury mortality, counseling about parent education on this subject, and first aid in emergencies. PMID- 9346018 TI - How do practice nurses see their role in childhood injury prevention? AB - OBJECTIVES: To assess the knowledge of unintentional injury epidemiology, the attitudes towards, and current practices in injury prevention among practice nurses. SETTING: Practice nurses employed by general practitioners in Nottinghamshire, United Kingdom. METHOD: A postal questionnaire was sent to all practice nurses on the Family Health Services Authority list (n = 322) with questions covering sociodemographic details, occupational details, unintentional injury epidemiology, attitudes towards the injury prevention activities suggested by a government report as part of the role of the primary health care team, and current practices in injury prevention. RESULTS: A response rate of 71.1% was achieved. More than 50% knew that unintentional injuries were the most common cause of death in childhood. A similar per cent knew the site of most fatal injuries in the under 1 and 5-16 year age groups. More than two thirds correctly identified a range of risk factors for unintentional injury. However, only two fifths of nurses believed they could be effective in preventing injuries. There were considerable gaps between attitudes and practice for most activities. The activities most commonly undertaken include displaying posters and leaflets (69.4%), giving advice on prevention (51.1%), and advice on first aid (45.0%) during injury consultations. CONCLUSIONS: Most practice nurses hold positive attitudes towards injury prevention activities, but fewer undertake these activities regularly. The activities most commonly undertaken employ an educational model. Further research is needed on the barriers to practice nurses undertaking more injury prevention work, the effectiveness of systems to overcome such barriers, and the effectiveness of these injury prevention activities. PMID- 9346019 TI - General practitioners' attitudes to child injury prevention in the UK: a national postal questionnaire. AB - OBJECTIVE: To survey the level of interest and involvement in child injury prevention among general practitioners and their practice teams, and to identify factors associated with current interest. DESIGN: Postal survey of a random sample of United Kingdom (UK) medical practitioners. SETTING: Medical practices throughout the UK. SUBJECTS: 957 general practitioners (50% of the total sample) who responded to the survey questionnaire. OUTCOMES: Answer to questions about role in injury prevention. RESULTS: Despite a response rate of only 50%, this study is the largest to examine the role of general practitioners in child injury prevention. Seven hundred and twenty five (77%) of the respondents considered injury prevention to be part of the general practitioner's role, but only 260 (28%) felt that they did enough in this area. Time was cited as the most significant limiting factor. Women doctors, rural practitioners, members of the Royal College of General Practitioners, and doctors with previous personal experience of serious accidents all had more positive attitudes to injury prevention as a routine part of their activities (p < 0.05). Practices providing first aid training for staff were also associated with an interest in injury prevention. The most appropriate times for offering prevention advice were thought to be during child health surveillance clinics and during treatment of an accident. CONCLUSIONS: Awareness about injury prevention opportunities might be improved by emphasising the roles of individual team members and by better addressing the training needs of the whole team. PMID- 9346020 TI - The Lidkoping Accident Prevention Programme--a community approach to preventing childhood injuries in Sweden. AB - OBJECTIVES: In Sweden about 100 children 0-14 years die from accidental injuries every year, roughly 40 girls and 60 boys. To reduce this burden the Safe Community concept was developed in Falkoping, Sweden in 1975. Several years later a second programme was initiated in Lidkoping. The objectives of this paper are to describe the programme in Lidkoping and to relate it to changes in injury occurrence. SETTING: The Lidkoping Accident Prevention Programme (LAPP) was compared with four bordering municipalities and to the whole of Skaraborg County. METHODS: The programme included five elements: surveillance, provision of information, training, supervision, and environmental improvements. Process evaluation was based mainly on notes and reports made by the health planners, combined with newspaper clippings and interviews with key people. Outcome evaluation was based on information from the hospital discharge registry. RESULTS: In Lidkoping there was an on average annual decrease in injuries leading to hospital admissions from 1983 to 1991 of 2.4% for boys and 2.1% for girls compared with a smaller decline in one comparison area and an increase in the other. CONCLUSION: Because the yearly injury numbers are small there is a great variation from year to year. However, comparisons over the nine year study period with the four border municipalities and the whole of Skaraborg County strengthen the impression that the programme has had a positive effect. The findings support the proposition that the decrease in the incidence of childhood injuries after 1984 could be attributed to the intervention of the LAPP. Nevertheless, several difficulties in drawing firm conclusions from community based studies are acknowledged and discussed. PMID- 9346022 TI - The New Zealand graduated driver licensing system: teenagers' attitudes towards and experiences with this car driver licensing system. AB - OBJECTIVES: This study examined the attitudes of teenagers towards the New Zealand graduated driver licensing system (GDLS), and the extent to which it affected them. METHOD: Teenagers, who are members of a longitudinal study of a birth cohort, were interviewed at 15 years of age when the GDLS was first introduced and before they had begun licensure, and again at 18 years of age after they had experience with this licensing system. RESULTS: At both ages the majority (over 70%) agreed with the driving restrictions imposed by this system. After experience with the restrictions, however, significantly more reported being affected a lot by them, than had expected to be at age 15. This was especially true of the restrictions on the carrying of passengers and the night time curfew (10 pm - 5 am). However, few reported that they were affected by the alcohol restriction. Sixty eight per cent of those with a graduated licence reported breaking at least one of the conditions, most frequently carrying passengers. Very few were penalised by the police for this. CONCLUSIONS: Generally these young drivers were positively disposed towards the driving restrictions, but noncompliance was common. A full evaluation of all aspects of this licensing system is recommended. PMID- 9346021 TI - Injury surveillance in children--usefulness of a centralised database of accident and emergency attendances. AB - OBJECTIVE: To assess the usefulness of a centralised injury database in monitoring progress towards nationally set health targets for the reduction of childhood injuries. SETTING: West Glamorgan County, Wales. METHODS: Analysis was undertaken of data held in the West Glamorgan injury database which amalgamates population data with data from the three hospital units covering a population of 370,000. All first attendances due to a new injury in children aged 0-14 occurring in 1993 were analysed, with subgroup analysis for injuries occurring in the home and injuries resulting in fractures. Standardised injury ratios were compared with the distance travelled, car ownership, and Townsend index of deprivation at the ward level, using multiple linear regression. RESULTS: A total of 10,117 first time visits due to injuries were recorded, representing a rate of 182 injuries/1000 children aged 0-14 in West Glamorgan County. Distance from home to the accident and emergency departments was inversely correlated with total injury attendances, and injuries occurring at home, but not with injuries resulting in fractures. Visit rates for any type of injury were not associated with local car ownership rates or deprivation indices. CONCLUSIONS: Proximity to accident and emergency departments is a strong determinant of the use of the service by children with overall injuries, and injuries occurring at home. The lack of a significant association between travel distance and injuries resulting in fractures suggests that it is more meaningful to use a centralised database of accident and emergency department attendances to monitor the more severe spectrum of childhood injuries in assessing progress towards national targets for their reduction. The absence of an association between severe injuries and local socioeconomic factors suggests that national targets for the reduction of socioeconomic differentials in childhood injuries may need to be reassessed. These databases are also useful in generating information to direct preventive strategies and to target resources to areas of greatest need. PMID- 9346023 TI - Engineering safety on the road. PMID- 9346024 TI - Cost effective ways to make walking safer for children and adolescents. PMID- 9346025 TI - Falls from heights: a childhood epidemic in an urban area. 1971. PMID- 9346026 TI - Children can't fly: a program to prevent childhood morbidity and mortality from window falls. 1977. PMID- 9346027 TI - CAPFSA--facing the problem of childhood injury in South Africa. Child Accident Prevention Foundation of Southern Africa. PMID- 9346028 TI - Making injury prevention a top social issue: the Canadian Injury Prevention Foundation. PMID- 9346030 TI - Epidemiology, and what then? PMID- 9346029 TI - Children and bicycles. PMID- 9346031 TI - Epidemiology and product safety--opportunities and limitations. PMID- 9346032 TI - Where have all the programmes gone? PMID- 9346034 TI - Cautionary notes on teaching water safety skills. PMID- 9346033 TI - Drowning prevention in children: the need for new strategies. PMID- 9346035 TI - 'What I said' versus 'what you heard': a comparison of physicians' and parents' reporting of anticipatory guidance on child safety issues. AB - OBJECTIVE: Unintentional injuries are the number one cause of death for infants. Many of these injuries could be prevented if parents took additional safety precautions. In this study physicians' and parents' perspectives regarding the part that physicians play in educating first time parents about child safety issues were compared. METHODS: All pediatricians and family physicians in London, Ontario were surveyed by mail (68% return rate) regarding their practices, attitudes, and beliefs related to parent education about child safety issues. A sample of 114 first time mothers, including 38 each with 6, 12, and 18 month old infants, completed a telephone interview. All parents had physicians who had returned questionnaires. RESULTS: There was good correspondence between parents' and physicians' judgments about the safety issues most often covered, and what role physicians should adopt regarding parent education about child safety issues. In addition, they both agreed that parents seldom seek out safety information by asking questions. Relative to parent reports, however, physicians significantly overestimated the time they spent on safety issues and the degree of their direct involvement in communicating this information. The best predictor of time spent by physicians on safety issues was their rating of the importance of assuming the role of parent educator. The best predictor of parents asking questions about child safety was their rating of the adequacy of physicians' responses to previously asked questions. CONCLUSIONS: The results suggest that both physicians and parents contribute to undermine communication about child safety during well-baby visits. PMID- 9346036 TI - Water safety training as a potential means of reducing risk of young children's drowning. AB - OBJECTIVES: To determine the effects of training in swimming and water safety on young preschool-children's ability to recover safely from a simulated episode of falling into a swimming pool. DESIGN: Randomized trial of 12 or eight weeks' duration water safety and swimming lessons for children 24 to 42 months old. OUTCOME MEASURES: Swimming ability, deck behavior, water recovery, and swimming to side after jumping into pool were measured before, during, and after the training program. RESULTS: 109 children completed the study (61 in the 12 week group, 48 in the eight week group). The average age was 34.2 months, 54% were male. Swimming ability, deck behavior, water recovery, and jump and swim skills improved over baseline levels in both groups. By the end of training, the 12 week group improved more than the eight week group only in swimming ability. Improvements in water recovery and jump and swim skills were associated positively with changes in swimming ability. CONCLUSIONS: Swimming ability and safety skills of young preschool children can be improved through training. Such programs may offer some protection for children at risk of drowning and there was no indication that this program increased the risk of drowning. However, pool fencing, other barriers around water, and parental supervision still remain the most important prevention strategies to reduce drowning in young children. PMID- 9346037 TI - Surveillance of pediatric injury hospitalizations in Southern California. AB - OBJECTIVES: This study was designed to determine the incidence and causes of injury hospitalizations/fatalities to children less than 15 years of age. SETTING: Central Orange County, California. DESIGN: Cases were identified through a population based hospital and coroner's office surveillance system. SUBJECTS: The sample consisted of children 0-14 years of age who were residents of the study area and sustained an injury between 1 January 1991 and 31 December 1992 resulting in hospitalization or death. RESULTS: Over the two year study period, 1361 children 0-14 years of age were hospitalized or died as a result of injury. This represents a crude annual injury rate of 318/100,000 children. Rates were highest for children less than 5 years--this age group sustained the highest rate for eight of nine specific causes of injury. Falls were the leading cause of hospitalizations for all ages. Pedestrian injuries were more common among children 1-4 years and 5-9 years, while bicycle injuries were more common among older children. CONCLUSIONS: This study, one of the first population based studies in a Southern California urban/suburban community, found lower rates of injury hospitalization than studies conducted over a decade ago. These lower rates may reflect changes in hospitalization trends and/or injury prevention programs. Comparisons with more recent studies in inner city communities in the north east also show regional differences in rates and causes. Injury prevention efforts should particularly address the higher injury rates among children less than 5 years of age. This study also illustrates the need for regional and local data to guide injury control. PMID- 9346038 TI - Cycling to school--a significant health risk? AB - OBJECTIVES: The risk of injury to children riding bicycles has been previously documented. However, the specific risk arising from the use of bicycles as a mode of transportation to and from school is unknown. This study examines the incidence of bicycle related injuries among school age children. METHODS: A comprehensive prospective injury registration system was established in Stavanger, Norway. Data were obtained from this system to identify bicycle related injuries occurring from 1990-3 to children aged 10-15. The incidence of injuries was computed for two groups of children: (1) children cycling to school and (2) children cycling for other purposes. RESULTS: 352 children received medical treatment for bicycle related injuries, 12.6/1000 bicycle riders; 108 (30%) of the 352 children were injured while cycling to or from school. The incidence of bicycle related injuries was significantly higher for boys than girls. Seventy seven per cent of the injuries occurred in a non-collision accident, 9% in a collision with another bicycle, and 14% in a collision with a motor vehicle. Twenty per cent of the injured children sustained upper head injuries and 13% required inpatient treatment. Average maximum abbreviated injury severity (MAIS) score was similar for the injuries sustained during travel to/from school and other injuries. CONCLUSIONS: Bicycle related injuries occurring during travel to or from school are a significant contributor to the total incidence of bicycle related injuries. Increased attention among parents, school officials, public health officials, and medical professionals should be paid to this health risk. PMID- 9346039 TI - Adult accompaniment and the risk of pedestrian injury on the school-home journey. AB - STUDY OBJECTIVE: To quantify the effect of adult accompaniment on the risk of pedestrian injury on the school-home journey. DESIGN: A community based case control study. SETTING: The Auckland region of New Zealand. PARTICIPANTS: Cases (n = 54) were all children killed or hospitalized as a result of a pedestrian injury occurring on the school-home journey between 1 January 1992 and 1 March 1994. The response rate for the case group was 98%. Controls (n = 157) were a random sample of all children who walk to and from school in the study region. The response rate for the control group was 100%. MAIN RESULTS: Adult accompaniment on the school-home journey was associated with a reduced risk of injury (odds ratio (OR) 0.36, 95% confidence interval (CI) 0.04 to 1.66). This effect persisted after controlling for age, sex, and socioeconomic status (OR 0.31, 95% CI 0.07 to 1.49). CONCLUSIONS: Adult accompaniment on the school-home journey may have the potential to significantly reduce child pedestrian injury rates. The effect of adult accompaniment may have important implications for the interpretation of child pedestrian exposure studies. PMID- 9346040 TI - Motor vehicle occupant injuries in children 2 years and younger: a comparison between Western Australia and New South Wales 1982-92. AB - OBJECTIVES: To compare the age specific rate of passenger injury and associated restraint use for children 2 years and younger in the state of Western Australia (WA), with the state of New South Wales (NSW), Australia for the period 1982-92. SETTING: The states of WA and NSW, Australia. METHODS: A descriptive retrospective study of child passenger injuries in WA and NSW was conducted for the period 1 January 1982 to the 31 December 1992. The data provided information about the injured child, such as sex and restraint use, the driver, vehicle, and collision factors, such as time and posted speed limit. RESULTS: A total of 2280 children aged 0 to 2 years were injured in motor vehicle collisions during the study period. Of these children, 653 were from WA and 1627 from NSW. Both the injury and mortality rates were higher in WA compared with NSW over the study period. However 80% and 79% of child passengers injured in WA and NSW, respectively, were restrained at the time of injury. Thus both the sex of the driver and the year the motor vehicle was manufactured best predicted the likelihood of a child being restrained. CONCLUSIONS: As at 1992, WA's population age specific passenger injury rate for children 0-2 years was more than twice the rate in NSW. The comparable rate of reported restraint use by injured children 0 2 years in WA and NSW suggests that non-use of restraints cannot be singled out as the most likely cause of WA's comparatively high rate of injury. It is difficult to determine whether the disparity in rates could be explained by the child passenger's exposure to crash risk factors, as little is known about child passenger levels of exposure to these factors. Further research is needed to address this issue. PMID- 9346042 TI - Bicycle ownership, use, and injury patterns among elementary schoolchildren. 1971. PMID- 9346043 TI - Kidsafe Australia. PMID- 9346044 TI - Tractors, motorcycles, ATVs: inconsistencies in legislation for child safety. Examples from New Zealand. PMID- 9346045 TI - Child and adolescent injury control: what to do when the conference is over. PMID- 9346041 TI - Can we prevent accidental injury to adolescents? A systematic review of the evidence. AB - OBJECTIVES: As part of the Department of Health strategy The Health of the Nation, a systematic review of published and unpublished literature relating to the effectiveness of interventions in reducing accidental injury in the population aged 15-24 years was carried out. METHODS: The literature was reviewed under the standard setting headings of road, work, home, and sports and leisure, and graded for quality of evidence and strength of recommendation using a scale published in the UK national epidemiologically based needs assessment programme. RESULTS: The most effective measures appear to be legislative and regulatory controls in road, sport, and workplace settings. Environmental engineering measures on the road and in sports have relatively low implementation costs and result in fewer injuries at all ages. There is little evidence that purely educational measures reduced injuries in the short term. Community based approaches may be effective in all age groups, and incentives to encourage safer behaviour hold promise but require further evaluation. The potential of multifactorial approaches seems greater than narrowly based linear approaches. CONCLUSIONS: Few interventions to reduce injury in adolescents have been rigorously evaluated using good quality randomised controlled trials, and where such evidence is available, fewer have been shown to be definitely worthwhile. Many studies relied on surrogate measures rather than actual injury rates, and substantial issues relating to the efficacy or implementation of preventive measures in adolescent and young adult populations remain unresolved. PMID- 9346046 TI - Parental supervision: a popular myth. PMID- 9346047 TI - Is there more to parental supervision than political incorrectness? PMID- 9346048 TI - Annual incidence of unintentional injury among 54,000 children. AB - OBJECTIVE: To enhance the case definition of unintentional injuries in childhood by applying an objective severity measure to fatal and non-fatal cases. DESIGN: A descriptive prospective epidemiological study of a defined resident childhood population (< 16 years of age) for a one year period, 1990. SETTING: Newcastle upon Tyne, England. Child population estimate for 1990 was 54,400. SUBJECTS: Resident children who died, were admitted to local hospitals, or attended local accident and emergency departments. OUTCOME MEASURES: Using recognised severity scoring systems (for example the injury severity score, trauma score) injuries were classified as severe, moderate, or mild. RESULTS: There were six deaths, 904 admissions, and 11,682 accident and emergency department attendances. All deaths, 25% of admissions, and 1% of accident and emergency attenders were classified as severe. The underlying determinants of severe injuries are different than those for all other injuries (for example age, social class). A comparison with a local survey in 1986 showed a 26% rise in hospital admissions, but no significant rise in the frequency of severe or moderately injured children. Comparisons with other international data showed higher rates of injury admissions and attendances for England, but no significant differences in the frequency of severe injuries. CONCLUSIONS: Objective severity scoring enhances the case definition of unintentional injuries in childhood by allowing for the identification, and, therefore, the more reliable ascertainment of severely injured children. This more completely ascertained set of population cases increases the accuracy of comparisons of injury frequency over time and by place, and, in addition, enhances our basic understanding about the epidemiological characteristics of childhood unintentional injury. PMID- 9346049 TI - Incidence and distribution of injury among schoolchildren aged 11-15. AB - OBJECTIVES: To measure the incidence and age and sex distribution of self reported experience of injuries in the preceding 12 month period among a representative national sample of Scottish schoolchildren and to validate the findings against other data sources. DESIGN: Self completed questionnaire administered in schools, April-June 1994. SUBJECTS: 4710 pupils aged 11, 13, and 15 years drawn from a representative sample of 270 classes with returns from 224 classes (83.2% completion rate). OUTCOME MEASURES: Number, type, site, and severity of injuries reported. RESULTS: 41.9% of pupils reported a medically attended injury, with injury incidence significantly higher in boys than in girls. Using the abbreviated injury scale (maximum abbreviated injury score) one third of injuries were either moderate or severe. CONCLUSION: The incidence and distribution of self reported injury is consistent with estimates based on other data sources thus confirming the utility of this method of injury surveillance in this age group. PMID- 9346050 TI - Teaching safety: evaluation of a children's village in Maryland. AB - OBJECTIVES: The purpose of this study was to evaluate Children's Village, a life safety education facility for children. SETTING: The study took place in Washington County, Maryland, a rural county. METHODS: Eight elementary schools with 20 second grade classrooms (410 students aged 7 and 8) were selected to participate. Using a quasiexperimental design, tests were administered to two cohorts of children before (pretest) and after (post-test) they attended the Children's Village during 1993-4. Parent and teacher surveys were also completed after the program. RESULTS: Among children who attended in December 1993-January 1994, there was a significant improvement in average test scores between the pretest (58% correct) and post-test (78%). Among children who attended in April 1994, there also was a significant improvement in test scores between pretest (74%) and post-test (85%). Among parents, 70% reported that their child learned a great deal at Children's Village and 33% reported having made changes in their home as a result. The parent survey also revealed that 25% of children and 35% of adults did not always wear their seat belts, and 74% of children did not always wear bicycle helmets. Teachers' responses to the program were generally positive. CONCLUSIONS: Children's Village brought together an extensive network of community leaders, parents, and teachers dedicated to safety education of children. The curriculum had a positive impact on children's knowledge and, to a lesser extent, on parents' safety practices. Program impact could be enhanced by more emphasis on automobile restraints and helmets (behaviors that parents reported were not consistently practiced) and by expanding the village services to parents as well as children. Others considering creating similar programs need to identify community leaders willing to commit the time, effort, and resources required to develop and sustain such programs. PMID- 9346051 TI - Effects of North Carolina's mandatory safety belt law on children. AB - OBJECTIVES: To assess the effect of the North Carolina law mandating that all front seat passengers use a safety belt on children 4 through 15 years of age. METHODS: North Carolina collision reports, completed by local police or the state highway patrol for crashes with greater than $500 worth of damage, were analyzed using time series analysis on the monthly percentage of deaths and serious injuries between January of 1980 and February of 1994. RESULTS: Following the 1985 implementation of the law, children 4 to 15 years of age experienced a 42% decline in deaths and serious injuries. CONCLUSIONS: The mandatory safety belt law in North Carolina has been associated with a decline in deaths and serious injuries. Additional research in needed to assess the seat belt behaviors of this age group as well as the specific effects of seat belt use using outcome measures more precise than those available in police crash reports. PMID- 9346052 TI - Information through television: does it promote child safety? AB - OBJECTIVES: First, to evaluate whether a local campaign to prevent childhood injuries increased parents' inclination to follow eight television programmes broadcast nationwide, and second, to assess whether parents reached by a local campaign benefitted more from the television programmes than those not reached by the campaign. METHODS: Before the television programmes were broadcast, all families with preschool children living in a typical Swedish municipality (the intervention area) received a letter from the head of the child health services encouraging them to watch the programmes. The local campaign also included face to-face information and advice on childhood injuries at all day care centres and child health centres in the intervention area. After all the programmes had been broadcast, telephone interviews were conducted with one parent from 77% of all 1699 households with at least one preschool child in the intervention area, and with 87% of a random sample of 144 parents from other, similar municipalities. RESULTS AND CONCLUSIONS: The local campaign increased parents' inclination to follow the programmes. No significant association was found, however, between the number of programmes followed and measures undertaken in the homes as a direct consequence of the programmes. Nor was a significant association found between the number of programmes viewed and parents' attitudes towards risks. A local campaign may increase parents' awareness of information provided by the mass media on childhood injuries. PMID- 9346053 TI - Purchasing a cycle helmet: are retailers providing adequate advice? AB - OBJECTIVES: The aim of this study was to examine the selling of cycle helmets in retail stores with particular reference to the adequacy of advice offered about the fit and securing of helmets. METHODS: All 55 retail outlets selling cycle helmets in Christchurch, New Zealand were studied by participant observation. A research entered each store as a prospective customer and requested assistance to purchase a helmet. She took detailed field notes of the ensuing encounter and these were subsequently transcribed, coded, and analysed. RESULTS: Adequate advice for helmet purchase was given in less than half of the stores. In general the sales assistants in specialist cycle shops were better informed and gave more adequate advice than those in department stores. Those who have good advice also tended to be more good advice also tended to be more active in helping with fitting the helmet. Knowledge about safety standards was apparent in one third of sales assistants. Few stores displayed information for customers about the correct fit of cycle helmets. CONCLUSIONS: These findings suggest that the advice and assistance being given to ensure that cycle helmets fit properly is often inadequate and thus the helmets may fail to fulfil their purpose in preventing injury. Consultation between retailers and policy makers is a necessary first step to improving this situation. PMID- 9346054 TI - Cross cultural child injury prevention awareness. AB - PURPOSE: The purpose of this study was to determine how injury prevention awareness of children ages 3, 4, and 5, based on recognition of hazards in pictures differs in the United States, Belgium, East Germany and West Germany. METHODS: Children from these four countries were presented with 10 different pictures. Each picture represented a common injury producing situation to which children are exposed in traffic, home, and recreation. RESULTS: Results indicate that for pictures relating to home hazards, less than 22% of children from Belgium (21.5%), West Germany (4.7%), and the United States (20.3%) clearly recognized the essential hazards in the pictures, whereas over 40% of the East German children clearly recognized these dangers. A higher proportion of the children from all countries recognized the traffic hazards. Only 23.9% of all children had a clear recognition of the playground situations. The child's age had a bearing on ability to recognize hazards overall. CONCLUSIONS: Children need to be provided with better injury prevention education at an early age, especially those from West Germany. PMID- 9346055 TI - The Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP) in the UK: a pilot study. AB - OBJECTIVES: To assess the feasibility, strengths and weaknesses, and preventive utility of the Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP) in a paediatric setting in the UK. DESIGN: Implementation and operational evaluation of CHIRPP. SETTING: A paediatric accident and emergency department in Glasgow, Scotland, UK. METHODS: CHIRPP forms were used to collect and analyse data on the circumstances, mechanisms, and types of injuries in 2516 children (age range 0-13 years) presenting to the accident and emergency department over the period of 1 April 1993 to 31 January 1995. The strengths and weaknesses of CHIRPP were assessed by direct observation, discussion with staff, operation of the CHIRPP software, and scrutiny of the output. RESULTS: After initial technical problems, CHIRPP ran smoothly. Although parental compliance was high, staff compliance was low, and this resulted in a low capture rate. Tabulations indicated the potential of the system for identifying both hazardous environments and vulnerable population subgroups at whom specific preventive measures can be targeted. Specific proposals for enhancing the efficiency and preventive utility of CHIRPP in this setting were formulated. CONCLUSIONS: CHIRPP offers hospitals, public health departments, and government agencies in the UK a promising tool for planning national, regional, and local injury prevention. PMID- 9346056 TI - Dog bites: how big a problem? AB - OBJECTIVE: To estimate the magnitude of the dog bite problem in the US. METHODS: Data on dog bites were gathered as part of a 1994 national telephone survey of 5,238 randomly dialed households. Data were weighted to provide national estimates. RESULTS: The weighted total number of dog bites was 4,494,083 (estimated incidence = 18/1,000 population); of these, 756,701 persons sustained bites necessitating medical attention (incidence rate = 3/1,000). Children had 3.2 times higher medically attended bite rates than adults (6.4/1,000 children v 2/1,000 adults). CONCLUSIONS: More attention and research needs to be devoted to the prevention of dog bites. Potential prevention strategies include: educational programs on canine behavior, especially directed at children; laws for regulating dangerous or vicious dogs; enhanced animal control programs; and educational programs regarding responsible dog ownership and training. Unfortunately, the relative or absolute effectiveness of any of these strategies has not been assessed. Continuing surveillance for dog bites will be needed if we are to better understand how to reduce the incidence of dog bites and evaluate prevention efforts. PMID- 9346057 TI - Theory and methods of epidemiologic study of home accidents. 1963. PMID- 9346058 TI - Limitations of child injury data from the CPSC's National Electronic Injury Surveillance System: the case of baby walker related data. AB - OBJECTIVES: The US Consumer Product Safety Commission's National Electronic Injury Surveillance System (NEISS) is a primary source for children's consumer product injury surveillance data in the US. Differing interpretations of the emergency department based NEISS baby walker data by various parties prompted this detailed examination, reclassification, and analysis of the NEISS data to explain these discrepancies. METHODS: Case selection was performed by searching the NEISS 1982-91 database for the baby walker product code and various text strings for children less than 24 months old. False negative and false positive cases were identified and reclassified. Adjusted population rates were computed and the types and locations of hospitals contributing to the sample were examined. RESULTS: One per cent false positive and 4% false negative misclassification rates were observed. In 1991, two children's hospitals reported 14% of the baby walker related injuries, though these hospitals made up just 2% of the sample frame. Through random allocation, one state currently contains four acute care hospitals and the only two children's hospitals reporting to the NEISS system. These six hospitals contributed 18% of the walker cases whereas the state represents only 3% of the US infant population. CONCLUSIONS: Misclassification in NEISS baby walker reports is minimal, with false negatives outweighing false positives. For trend analysis of product related injuries at the frequency of occurrence observed for baby walkers, NEISS suffers from low sensitivity due to sampling error. For children's injuries, NEISS' estimates have been affected by children's hospitals coming in and out of the sample and currently reflects a random geographic imbalance because one state contributes both of the reporting children's hospitals. To overcome these problems improved multiple product coding, a unique baby walker code, and stratification of children's hospitals in an enlarged NEISS sample is recommended. PMID- 9346059 TI - Validity of self reported data on injury prevention behavior: lessons from observational and self reported surveys of safety belt use in the US. AB - OBJECTIVES: To examine the validity of self reported data on safety belt use and to consider the implications for research on injury prevention behaviors. METHODS: 1992 and 1993 self reported data on safety belt use were obtained from the Behavioral Risk Factor Surveillance System and observational data were obtained from the National Highway Traffic Safety Administrations for 49 states in 1992 and 50 states in 1993. The ratio of self reported to observed belt use was calculated for each state, and linear regression models were used to examine the association between the two methods. RESULTS: There was variation between states, but the overall median ratio of self reported to observed safety belt use was 1.05 in 1992 (interdecile range 0.87-1.36) and 1.02 in 1993 (interdecile range 0.87-1.31). Self reports were substantially higher in southern states and in states with the lowest levels of observed use. Linear regression models indicated a moderately strong association between state estimates using both methods. For every percentage point increase in self reported data in 1993, observed safety belt use increased by 0.95 percentage point. CONCLUSIONS: In the aggregate, self reported estimates were only 2% to 5% higher than observed estimates. This is a substantial improvement from previous studies. This is probably due to the increased prevalence of safety belt use and the declining effects of social desirability on self reported use. In general, the validity of self reported estimates of socially desirable injury prevention behaviors will be higher when the actual prevalence of the behavior is higher, but lower when this is not true. PMID- 9346060 TI - SAFE KIDS Canada. PMID- 9346061 TI - Limitations of NEISS child injury data. PMID- 9346062 TI - Prevention is the key. PMID- 9346063 TI - Remarks from the Haddon Memorial Plenary Session. PMID- 9346064 TI - Health status measurement: the special case of children and youth. PMID- 9346065 TI - An update from the business meeting in Melbourne. PMID- 9346066 TI - Injury control in developing nations: what can we learn from industrialized countries? PMID- 9346067 TI - Injury control in developing countries: context more than content is crucial. PMID- 9346068 TI - Injury mortality among children and teenagers in the United States, 1993. PMID- 9346070 TI - Injuries and their relation to potential hazards in child day care. AB - OBJECTIVES: To prospectively determine the incidence rate of injuries that required medical attention among children in day care and to identify possible hazards related to these injuries. SETTING: King County, Washington. METHODS: Prospective cohort study of children in a sample of licensed day care facilities. RESULTS: From 1 July 1992 to 30 June 1993, 53 medically attended injuries were reported by 133 day care sites; incidence rate 1.9 per 100,000 hours of day care attendance. The rate of injury in 91 small family day care homes was essentially the same as that in 42 larger day care centers; relative rate 1.0 (95% confidence interval 0.6 to 1.9). Injuries that required sutures accounted for 39% of the cases, while 17% required a cast, splint, or sling. No child was hospitalized. Sixty nine sites were inspected and all had potentially correctable physical hazards, with a median of 15 hazards per site (range 7 to 26). These potential hazards had little relationship to the risk of injury and a case-by-case review identified only two injuries that might have been prevented by a more energy absorbent playground surface. CONCLUSIONS: The incidence of medically attended injuries found in this study is consistent with other studies from the United States. Most injuries were minor and had little relation to physical hazards at day care locations. PMID- 9346069 TI - Height and surfacing as risk factors for injury in falls from playground equipment: a case-control study. AB - OBJECTIVES: Despite the widespread promotion of safety standards no epidemiological studies have adequately evaluated their effectiveness in preventing injury in falls from playground equipment. This study evaluated the effectiveness of the height and surfacing requirements of the New Zealand standard for playgrounds and playground equipment. SETTING: Early childhood education centres and schools in two major cities in the South Island of New Zealand. METHODS: Data were collected on 300 children aged 14 years or less who had fallen from playground equipment. Of these, 110 (cases) had sustained injury and received medical attention, while 190 (controls) had not sustained injury requiring medical attention. RESULTS: Logistic regression models fitted to the data indicated that the risk of injury being sustained in a fall was increased if the equipment failed to comply with the maximum fall height (odds ratio (OR) = 3.0; 95% confidence interval (CI) 0.7 to 13.1), surfacing (OR = 2.3; 95% CI 1.0 to 5.0), or safe fall height (OR = 2.1; 95% CI 1.1 to 4.0) requirements. Falls from heights in excess of 1.5 metres increased the risk of injury 4.1 times that of falls from 1.5 metres or less and it was estimated that a 45% reduction in children attending emergency departments could be achieved if the maximum fall height was lowered to 1.5 metres. CONCLUSIONS: Although the height and surfacing requirements of the New Zealand standard are effective in preventing injury in falls from playground equipment, consideration should be given to lowering the maximum permissible fall height to 1.5 metres. PMID- 9346071 TI - Can child fatalities in house fires be prevented? AB - OBJECTIVES: To analyse all child deaths in house fires in Scotland between 1980 and 1990. METHODS: Retrospective study of all child house fire fatalities based on the 'sudden death' investigation instigated by the procurator fiscal in whose jurisdiction the death occurred. The necropsy, toxicology, police, and fire brigade reports were examined in each case. RESULTS: There were 168 child deaths occurring in 118 house fires. In the 0-5 years age group 40% of deaths occurred in fires started as a direct result of the actions of children. The careless disposal of smoking materials was the most frequent cause of fatal fires killing older children. Upholstery and bedding were common materials of first ignition, accounting for over half the incidents. The majority of children were dead before the arrival of the emergency services and most died as a result of the inhalation of smoke. CONCLUSIONS: This survey emphasises the importance of 'self escape' which, particularly in the case of young children, requires the assistance of adult carers. The number of fires started as a result of children playing with sources of ignition raises important questions of supervision and the provision of a safe environment. There is, we contend, a need to highlight the importance of individual behaviour and responsibility while recognising the need to develop measures that are relevant to, and effective in, a particular socioeconomic context. PMID- 9346072 TI - Airgun injuries in New Zealand, 1979-92. AB - OBJECTIVES: To describe the epidemiology of serious airgun injury in New Zealand. METHODS: Cases were selected from the New Zealand Health Information Service's hospital inpatient morbidity data files for the period 1979 to 1992 inclusive. RESULTS: There were 718 airgun related injuries resulting in 1.56 injuries/100,000 population/year. Males and 10-14 year olds had higher than average rates of injury. The majority of the incidents were unintentional. There has been a marked decline in injury rates since 1989. CONCLUSIONS: Airgun injuries, while not as serious as powder firearm injuries, account for a significant personal and societal burden. The results suggest that strategies aimed at controlling these injuries, especially those pertaining to children, are in need of review. PMID- 9346074 TI - Regional variation in homicide rates of infants and children. AB - GOAL: This study examines regional correlates of homicide rates for infants and children for the states of America. SAMPLE: The sample consists of all homicide victims in the 48 continental, contiguous states of America in both 1980 and 1990 from the ages of 0-14. METHODS: The homicide rates of infants aged 0-1 and children aged 1-4 and 5-14 were correlated with social indicators for the American states in 1980 and 1990. RESULTS: In 1980, children aged 1-14 had higher homicide rates in the more generally violent, urban, and socially disorganized states. However, no correlates for the homicide rate of infants were identified, suggesting that different theories may be required to explain their deaths. The analyses were repeated using data for 1990, at which time crime rates and fewer medical facilities were more weakly associated with the homicide rate of infants. CONCLUSIONS: The results suggest that different sociological theories may be required to account for the regional variation in the homicide rates of infants from those used for explaining the variation in the rates of children. PMID- 9346073 TI - Hazards of baby walkers in a European context. AB - OBJECTIVES: To identify conditions related to baby walker injuries in a Greek population. DESIGN: Analysis of all baby walker related injuries recorded during a 12 month period by the childhood injury surveillance system established in one of the two teaching hospitals for children serving the population of Athens. SETTING: Emergency clinics of A Kyriakou Children's Hospital in Athens, Greece. SUBJECTS: 49 babies with baby walker related injuries brought to the emergency clinics during the period May 1994 to April 1995. RESULTS: The incidence of these injuries was 16 per thousand person years of users, or 3.5 per thousand babies per year. More boys than girls were brought to the hospital for these injuries and the incidence density was highest during the ninth and 10th month of age. Falls from heights, particularly stairs, were the most frequent cause of baby walker related injuries, especially among younger babies. The majority of these injuries were of minor severity, but three babies had bone fractures and one had a second degree facial burn. Six babies required hospitalization and for seven others, a follow up visit was needed. The higher proportion of hospitalization among girls than boys raises the possibility that boys with minor injuries are more frequently brought to the hospital. CONCLUSIONS: Baby walkers impart a significant risk of injury from a consumer product that provides no clearly identifiable benefit. As most baby walker injuries happen on stairs, modifications in product design are required to reduce these injuries. Moreover, parents should be forcefully advised of the risks and predisposing conditions, if baby walkers are to be used at all. PMID- 9346075 TI - Golf cart related injuries in a North Carolina island community, 1992-4. AB - OBJECTIVES AND METHODS: The use of electric golf carts for roadway transportation is increasing in many regions of the United States, but injuries associated with the operation of these vehicles have not been previously described. In response to reports of golf cart related injuries in a North Carolina island community, we reviewed ambulance call report (ACR) information to identify and describe all injuries related to golf cart operation in this community in 1992-4. We also conducted telephone interviews with the subset of injured people who consented to be contacted. SETTING: Bald Head Island, North Carolina. RESULTS: Twenty two people were included in the case series, and 55% of these provided interview information to supplement ACR data. Fifty nine per cent of the 22 injured people were injured when they fell from a moving golf cart; of those injured in this manner, all with available information on seating position were passengers (rather than drivers). Eighty six per cent received immediate medical treatment at a mainland hospital. Thirty two per cent of injury incidents occurred among children aged 10 or younger. Forty per cent of injured adults were known to have been drinking alcohol before their injuries occurred, while alcohol was not known to have been involved in any of the children's injuries (in terms of drinking either by children or by accompanying adults). CONCLUSIONS: In settings where golf carts are used for road transportation, their users and traffic safety officials should be aware of potential safety hazards associated with the use of these vehicles, and installation of appropriate occupant restraints should be considered seriously. PMID- 9346077 TI - Lidkoping Accident Prevention Programme: what was the impact? PMID- 9346076 TI - Teenagers' attitudes towards bicycle helmets three years after the introduction of mandatory wearing. AB - OBJECTIVES AND SETTING: To address helmet wearing by 13-17 year olds this study posed the following research questions: 'Do education programs continue to be necessary even after the community wearing rate has increased?' and 'Are helmet laws more effective in encouraging wearing among certain age groups?' Victoria was the first place in the world to introduce bicycle helmet legislation. Experiences in Victoria therefore provide a good model for the introduction of similar legislation in other areas. This study is the first to examine teenagers' attitudes towards helmet wearing after the introduction of compulsory helmet wearing legislation. METHODS: A survey of 1240 year 9 and year 10 students, aged 13-17 years, from 14 secondary schools in the outer south eastern suburbs of Melbourne, was conducted in September 1993. Information about bicycle use, helmet wearing, and attitudes towards helmets was obtained by a self report questionnaire. RESULTS: Bicycles are a popular form of wheeled recreation/self transport among teenagers. 65% of teenagers reported that they owned a helmet but only one third wore a helmet the last time they rode a bicycle. Fewer than 25% of students always wore a helmet when they rode a bicycle, despite compulsory helmet wearing legislation. Major factors leading to teenagers not wanting to wear a helmet were appearance and comfort. Both safety considerations and parental pressures were factors that influenced a teenager to wear a helmet. CONCLUSIONS: The major areas that need to be addressed are low helmet wearing rates; the low priority given to safety issues compared with comfort and peer acceptance; an ignorance of the need for helmets in all riding situations; and a perception that the legislation would not be enforced. PMID- 9346078 TI - The SF-36 health survey: a valid measure of changes in health status after injury. AB - OBJECTIVES: The aim of this study is to evaluate the criterion validity and responsiveness to changes over time of the Medical Outcome Study Short Form 36 (MOS SF-36) measure. METHODS: A consecutive sample of 775 patients 16 to 78 years treated for an unintentional injury at the hospital or emergency clinic in Drammen, Norway was selected for the study. Data about activity restrictions and health status measured by SF-36 were obtained by a postal questionnaire 6-10 weeks after the injury. A follow up survey was sent 24-28 weeks later to all who reported activity restriction at the time of the first survey. Fifty two of these replied (63%). RESULTS: 469 patients responded to the survey questionnaire and of these, 82 experienced some restriction of activity. These scored lower (p < 0.01) on all eight SF-36 health dimensions (physical functioning, social functioning, role limitation (physical), role limitation (emotional), bodily pain, mental health, vitality, and general health) than the 387 patients without activity restriction. Scores on physical functioning, social functioning, role limitation (physical), bodily pain, and vitality significant improved (p < 0.01) among the 52 patients who were followed up. Scores on the other dimensions, however, showed no significant changes over time. CONCLUSION: The MOS SF-36 appears to be a valid instrument, responsive to changes in health status over time among unintentionally injured adult people. Thus it may be possible to use the SF-36 to describe changes in health due to injury. The applicability of this or similar measures for injured children remains to be established. PMID- 9346080 TI - The exposure of young children to accident risk as pedestrians. AB - Pedestrian road accidents show a marked peak for children aged 5, 6 and 7 years with boys twice as involved as girls at these ages. Howarth et al (1974) described a framework in which measures of exposure were defined and related to the accident statistics to obtain estimates of absolute levels of risk for different categories of pedestrian in different traffic situations. The present paper describes a survey of children's exposure carried out to provide suitable data for this quantitative analysis. We interviewed a representative sample of Nottingham schoolchildren about their journeys in the previous 24 hours and recorded the number of roads crossed and the traffic densities of these roads. The measures of exposure obtained are presented in relation to the accompaniment of children on their journeys, the type of area in which they live, and time of day. Risk was assessed by relating exposure measures both to the national and local accident statistics. The analysis provides estimates of the risk to children of different ages and sex in their normal pattern of road crossing and in crossing roads of different traffic density and indicates that the accident statistics alone considerably underestimate the degree of risk to children under the age of eight. Interviews with a sample of the parents of the children suggest that children may provide a more accurate measure of their exposure than do their parents. PMID- 9346082 TI - A modest proposal. PMID- 9346081 TI - Safekids. PMID- 9346079 TI - Preventing childhood unintentional injuries--what works? A literature review. AB - AIM: The aim of this paper is to report on a systematic review of the world literature to provide information about the most effective forms of health promotion interventions to reduce childhood (0-14 years) unintentional injuries. The findings are of relevance to policy makers at a local or national level, to practitioners and researchers. METHODS: The relevant literature has been identified through the use of electronic databases, hand searching of journals, scanning reference lists, and consultation with key informants. RESULTS: Examples of interventions that have been effective in reducing injury include: bicycle helmet legislation, area wide traffic calming measures, child safety restraint legislation, child resistant containers to prevent poisoning, and window bars to prevent falls. Interventions effective in changing behaviour include bicycle helmet education and legislation, child restraint legislation, child restraint loan schemes, child restraint educational campaigns, pedestrian education aimed at the child/parent, provision of smoke detectors, and parent education on home hazard reduction. For the community based campaigns, the key to success has been the sustained use of surveillance systems, the commitment of interagency cooperation and the time needed to develop networks and implement a range of interventions. Education, environmental modification and legislation all have a part to play and their effect in combination is important. CONCLUSION: The design of evaluations in injury prevention needs to be improved so that more reliable evidence can be obtained. Better information is needed on process, so that successful strategies can be replicated elsewhere. There is also a need for literature reviews on effectiveness to be updated regularly and for their findings to be widely disseminated to policy makers, researchers, and practitioners. PMID- 9346083 TI - Postmarketing surveillance of injury countermeasures. PMID- 9346084 TI - Closing remarks from the Haddon Memorial Plenary Session. PMID- 9346085 TI - Child and adolescent injury control activities: reports from the field. PMID- 9346086 TI - Etiological studies of traumatic injury: are we measuring the right outcome? PMID- 9346087 TI - Risk taking disease. PMID- 9346088 TI - Children falling from a height in London. AB - OBJECTIVES: To determine the frequency and geographical distribution of children falling from a height in London and to suggest possible causes and preventative measures. METHODS: All relevant cases attended by the Helicopter Emergency Medical Service (HEMS) in a three and a half year period were reviewed and the locations related to the boroughs. The rates, per 1,000 resident children, were compared with socioeconomic indices for the boroughs concerned. In addition, a survey was undertaken of window fittings and maintenance in high rise flats close to one particular incident. RESULTS: A total of 90 incidents were attended involving 91 patients (64 male, 27 female) of whom five died. HEMS attends approximately one third of incidents involving serious trauma. In the study period the maximum frequency was 0.2 fallers per 1,000 resident children, occurring in three boroughs. In three boroughs there were no fallers. There was no overlap in socioeconomic indices between the five boroughs with the highest fall rates and the five with the lowest. The building survey found a high incidence of faulty window catches, a slow response rate for repair, and a lack of safety advice for residents. CONCLUSIONS: The frequency of falling is related to urban deprivation, poor maintenance, and lack of safety information. A combination of regulation and targeted education could substantially decrease deaths and injuries in children from this cause. PMID- 9346089 TI - A population based case-control study of agricultural injuries in children. AB - OBJECTIVES: To identify preventable risk factors related to agricultural injuries occurring to children on family farms. SETTING: A geographically defined central region of Wisconsin, USA with nearly 1800 family dairy farms. METHODS: A two year, population based incidence study of occupational injuries among farm residents was conducted. For cases, trained staff abstracted information on the nature, severity, and treatment of the injury from the patient's medical record. Staff also administered a telephone questionnaire to cases and controls, usually answered by parents. RESULTS: There were 60 cases of farm residents younger than 18 years who sought care for acute agriculture related injuries. Farms on which uninjured children lived served as controls (n = 102). Multivariate analyses of 16 different variables revealed three significantly related to injuries to children: hours worked per week (odds ratio (OR) = 1.05; 95% confidence interval (CI) = 1.01 to 1.08); presence of disabled safety device (OR = 2.64; 95% CI = 1.10 to 6.35); and feeding cows by grazing (OR = 0.22; 95% CI = 0.06 to 8.83). CONCLUSIONS: Interventions designed to reduce the risk of agricultural injuries to farm children should acknowledge the participation of children as productive workers on the farm. Although education has been the standard method for encouraging safe practices in farm work, additional approaches, such as limiting the number of hours a child works, avoiding the disabling of safety devices, and using specific methods of managing cows, should also be adopted to minimize injury risks to farm children. PMID- 9346090 TI - Safety measures taken by Norwegian mothers. AB - OBJECTIVES: To identify predictors of the adoption of safety measures by mothers of 2 year old children. SETTING: 26 municipalities in the county of Sogn and Fjordane, and four municipalities in the county of More and Romsdal in Norway. METHODS: Data was collected by questionnaires mailed to all mothers of 2 year olds in the 30 municipalities (response rate 70.7%, n = 1233). Information was obtained on socioeconomic variables, the child's injury history, adoption of safety measures, and variables describing mother's health related beliefs (parent health locus of control) and the value of health (health value). RESULTS: Income, municipality of residence, age of the mother, and marital status were significantly associated with the reported adoption of safety measures. High income and older, married mothers were positively associated with the adoption of safety measures. CONCLUSIONS: The significant effect of income on the adoption of safety measures, underlined by the fact that safety measures were less often adopted by young single mothers, may indicate that the implementation of structural measures such as loan schemes and subsidies, are necessary to increase the adoption of child safety measures. The lack of association between education and social cognitive beliefs, respectively, and the adoption of safety measures, offer less optimism for traditional health education initiatives. PMID- 9346091 TI - A statewide survey of hazards in child care centers. AB - OBJECTIVES: The purpose of this study was to determine adherence to selected recommended safety standards in North Carolina child care centers. METHODS: A self administered questionnaire eliciting information about safety practices in child care was mailed to a randomly selected sample of 409 North Carolina child care centers. RESULTS: One hundred and ninety five usable questionnaires were returned from child care centers in 75 counties. Results indicated that all of the standards included in the state's child regulations were being adhered to by at least 80% of the centers. However, adherence to recommended standards not included in the state's regulations was quite variable, with one standard implemented by less than 5% of the centers. The lowest rates of adherence were found for standards specifying that resilient surface material be used under playground equipment (4%) and that certain foods that may present a choking hazard to small children not be served (27%). CONCLUSIONS: Many hazards not addressed in North Carolina child care regulations are present in child care centers. Some safety standards are not adhered to due to lack of knowledge or limited resources. Inclusion of national standards in state child care regulations appears to reduce, but not eliminate, the likelihood of hazards being reported. Further research should include on-site inspections and attention to safety in family child care. PMID- 9346092 TI - Risk factors for childhood poisoning: a case-control study in Greece. AB - OBJECTIVES: To identify child or family related risk factors for unintentional childhood poisoning in Greece and to explore whether product specific poisonings might have special features that make them amenable to preventive interventions. SETTING: A case-control study was undertaken in Athens, Greece in 1995. Cases were 100 consecutive children brought with poisoning to the emergency clinics of the two university affiliated children's hospitals. For every case two age, gender, and hospital matched controls were chosen from among children brought to the outpatient clinics of these hospitals on the same date. METHODS: All children and their guardians were interviewed by the same person using a standard questionnaire that covered demographic, socioeconomic, behavioral, and past injury characteristics. Information was also obtained concerning type and conditions of poisoning for cases. Statistical analysis was undertaken by modeling the data using conditional logistic regression. RESULTS: Socioeconomic factors were not important risk indicators in these data but children living with other than both parents were at increased risk (odds ratio (OR) = 4.7, p = 0.08), as were children with a history of previous poisoning that required medical care (OR = 5.1, p = 0.05). Unintentional poisonings caused by chewing or swallowing cigarettes were concentrated in families where both parents were smokers. CONCLUSIONS: Absence of a parent appears to be associated with increased likelihood of childhood poisoning. The importance of product accessibility is underlined by the concentration of tobacco poisoning among children of parents who were both smokers. In the cultural context of this study, sociodemographic factors do not appear to represent demonstrable risk factors. Instead, control of childhood poisoning should be concentrated on safe packaging, storage, and disposal of potentially hazardous products. PMID- 9346094 TI - Developing a regional network for preventing injuries of children and adolescents: the Region X experience. PMID- 9346093 TI - Child pedestrian and bicyclist injuries: results of community surveillance and a case-control study. AB - OBJECTIVES: To describe the dimensions of childhood pedestrian and bicyclist injuries in Long Beach, California, and to identify risk factors for these injuries. POPULATION: Long Beach residents aged 0-14 years who were involved in an auto versus pedestrian or bicyclist incident that resulted in a hospital visit and/or police response, between 1 September 1988 and 31 August 1990. METHODS: Cases were identified retrospectively using hospital charts, police records, and coroner's reports; demographic, clinical, and situational information were abstracted from the same. A nested case-control study was conducted to examine the street environments where children were injured, and to identify environmental risk factors at these case sites. RESULTS: 288 children comprised the sample population. Midblock dart-outs emerged as the single most common type of incident. Most incidents happened on residential streets, but the risk of injury was greatest on larger boulevards, and tended to cluster by region within the city. Adjusted odds ratios show that case sites had a larger proportion of traffic exceeding posted speed limits, and were also four times more likely to be near a convenience store, gas station, or fast food store than control sites. CONCLUSIONS: The findings of this study suggest three possible routes for the prevention of childhood pedestrian and bicyclist injuries: education, law enforcement, and environmental modification. PMID- 9346095 TI - Evaluation of a systematic approach for identifying injury scenarios. Kids'n' Cars Teams. AB - OBJECTIVE: To assess the effectiveness of a new multidisciplinary method for reconstructing the causal sequences that lead to child pedestrian injuries. SETTING: Subjects were 5-12 year old residents of Chicago, Illinois, USA, presenting for care due to pedestrian injury at one pediatric trauma center. METHODS: The interactions of medical, child, psychosocial, and traffic factors contributing to the injury were analysed. For 142 cases, information about the victim, his/her family, the injury site, and the activities just before the injury, was used in a structured manner by a multidisciplinary team to produce injury scenarios. Each scenario comprised a list of contributing factors, an estimate of the importance of each, and a narrative description of the causal sequence leading to the injury event. Face validity was assessed by two outside teams that performed a structured review of a subsample of cases (n = 11). Reliability was evaluated by comparison of the results of parallel teams assessing the same cases (n = 14). Process consistency and bias were assessed by analysis of the correlations of factor-importance rating patterns between members and over time. RESULTS: The outside team's agreement scores were based on a 1-5 Likert scale; these showed a mean of 3.6 and median of 4.0. Parallel teams consistently showed agreement greater than 85% on global attributes of cases. Intraclass correlation coefficient scores showed fair or better agreement for all classes of contributors, and excellent agreement for more than one third. Rating pattern analyses showed strong agreement by team members. Agreement did not increase over the period of the study. CONCLUSIONS: This causal sequence reconstruction method has acceptable face validity, reliability, and internal consistency. Although labor intensive and thus costly, it can produce unique, rich information for understanding injury causation and for guiding the search for promising interventions. PMID- 9346097 TI - The Swedish National Safety Promotion Program. PMID- 9346098 TI - Hand searching Injury Prevention. PMID- 9346096 TI - Developmental risk factors for childhood pedestrian injuries. PMID- 9346099 TI - Misleading airbag alarms. PMID- 9346100 TI - Injury research and violence: what's our contribution? PMID- 9346101 TI - Bicycle helmets--are they up to standard? PMID- 9346102 TI - Bicycle helmets reduce head injuries and should be worn by all. PMID- 9346103 TI - A breakthrough in gun control in Australia after the Port Arthur massacre. PMID- 9346104 TI - Bicycle helmet use among American children, 1994. AB - OBJECTIVE: To estimate ownership and use of bicycle helmets among children in the US in 1994. METHODS: As part of a 1994 national telephone survey of 5,238 randomly dialed households, adult respondents reported data on bicycle helmet ownership and helmet use among 1,645 child bicyclists. Data were weighted to provide national estimates. RESULTS: It is estimated that 72.7% of children 5-14 year olds ride bicycles, that is, 27.7 million child bicyclists. Of the bicyclists, 50.2% have a helmet and 25.0% reportedly always wore their helmet when cycling. Reported helmet ownership and use increased with income and educational level and decreased with age. Among regions of the US, those with the highest proportion of states with helmet use laws in 1994 also had the highest proportion of helmet use among children. Among child bicyclists who had been seen by a health care provider in the preceding 12 months, 43.9% of those counseled to wear a bicycle helmet were reported to comply compared with 19.1% of those seen by a provider but not so counseled (p < 0.001). CONCLUSIONS: To meet the year 2000 objective of 50% of bicyclists wearing helmets, use among American children will have to double. Concerted and increased efforts to promote the wearing of bicycle helmets are necessary. PMID- 9346105 TI - Smoke alarm use: prevalence and household predictors. AB - OBJECTIVE: To determine the prevalence of smoke alarm use among families with children and to identify household factors that predict the absence of a smoke alarm. DESIGN: Cross sectional analysis of data collected in the September and November 1995 Omnibus Survey, conducted by the Office of Population Censuses and Surveys in the UK. SUBJECTS: A random sample of British households. Interviews were completed with 4,043 householders. The response rate was 78%. RESULTS: 29% of British households do not have a smoke alarm and smoke alarms were absent in 20% of households with children under 15 years. A smoke alarm was absent in 41% of privately rented homes compared with 17% of owner occupied homes. Living in private rental accommodation was the strongest household predictor of the absence of a smoke alarm (odds ratio = 3.25, 95% confidence interval 1.94 to 5.42). Householders who had heard of National Fire Safety Week or the TV smoke alarm advertising campaign were significantly more likely to have a smoke alarm. The apparent effect of these campaigns was greatest in families with children. CONCLUSIONS: Smoke alarm use has continued to increase but a substantial proportion of British homes still do not have smoke alarms. Homes at greatest risk of residential fire are the least likely to have an alarm. Health professionals may be able to increase smoke alarm use among families with children, by counselling families about the benefits of smoke alarms. They may also be effective in this regard by lobbying local councils, houseing associations, or private landlords to install alarms in all properties and by advocating for national legislation. PMID- 9346106 TI - Behavior and injury in urban and rural adolescents. AB - OBJECTIVES: This study investigates the consistency of factors associated with adolescent injury in separate urban and rural samples. SAMPLES: Adolescents, 11 17 years old, in public schools in urban and rural Maryland (n = 2,712). METHODS: Separate bivariate and logistic regression analyses were conducted for each sample to determine individual and environmental factors associated with major and minor injuries experienced in the previous year. RESULTS: Multivariate analyses revealed that, for both samples, the probability of a major injury was highest for boys and, among both boys and girls, for those who played several team sports. Among rural youth, other significant covariates of both major and minor injuries were a tendency to engage in risky behavior and to use alcohol. For urban youth, being white, carrying a weapon for protection, attending an unsafe school, and working for pay were also significant covariates. Interactions were important and complex. CONCLUSIONS: The consistency of predictive factors, such as multiple sports team participation and risky and aggressive behaviors in completely different physical environments, underscores the need to address the contexts of heightened injury risk that some adolescents create wherever they live by playing sports and/or behaving in an antisocial, aggressive manner. Moreover, the perception of lack of safety in schools and neighborhoods is associated with increased injury rates, suggesting the need for policy interventions to target social environments as well as behavior. PMID- 9346107 TI - Work patterns and occupational hazard exposures of North Carolina adolescents in 4-H clubs. AB - OBJECTIVES: This study documents sex differences in work patterns, injuries, and hazard exposures among adolescents in homes, farms, and other work sites. METHODS: 14 to 17 year old 4-H club members were asked to complete self administered questionnaires regarding their lifetime experience of work, hazard exposure, and injuries. RESULTS: Of 323 respondents, more than two thirds had ever worked paid jobs. Fifty seven per cent were injured during non-farm work and hazards were part of the non-farm work environment for 54% of the respondents. Males were more likely to work in hazardous conditions, including operating heavy equipment on farms or construction sites. Almost three quarters of the teens who worked on farms reported being injured there and 100% were exposed to at least one farm hazard. CONCLUSIONS: Adolescents perform jobs at homes, farms, or other work sites where they are exposed to numerous safety hazards. Prevention efforts should target specific hazards youths are exposed to rather than the general work site. PMID- 9346108 TI - Firearm ownership and storage practices in Pennsylvania homes. AB - OBJECTIVE: To determine the household prevalence of firearms in Pennsylvania, and describe the storage practices for these weapons. DESIGN: A statewide telephone survey of 3,620 Pennsylvania adults selected from households by random digit dialing in 1994. MAIN OUTCOME MEASURES: Firearm ownership and storage practices were computed by household characteristics using logistic regression. RESULTS: The prevalence of firearm ownership was 37% (95% confidence interval = 35.4 to 38.6). Ownership of firearms was significantly higher for white residents, households with annual income of $20,000 or more, those in rural counties, and those with children and adolescents. Of the households with firearms, 23% contained a single firearm, the majority of which were handguns (40%) or rifles (40%); 76% had two or more firearms, with 57% reporting one handgun or more and 83% reporting one rifle or more. Storage of firearms in 72% of households involved two or more of these barriers: (1) taken apart; (2) trigger lock applied; (3) kept in a locked place; (4) unloaded; (5) no other ammunition; (6) locked ammunition; 6% stored at least one of their firearms with none of these barriers. The strongest predictor of storing a firearm with fewer than two protective barriers was households with no children or adolescents. CONCLUSIONS: Firearms are present in a large number of Pennsylvania homes. Many of these homes also contain children. To reduce the potential risks of firearms, optimal methods of storage of firearms in the home need to be determined. PMID- 9346109 TI - A community based approach to bicycle helmet use counts. AB - OBJECTIVE: Bicycle helmet use has become an important measure of the effectiveness of bicycle safety programs and the effectiveness of helmet legislation. Accounts of analytical comparisons of observation site selection methods are scarce. This report addresses this gap by reporting the relative effectiveness and costs of two alternative approaches to the selection of observation sites for helmet use counts. METHODS: The community based (COBA) method of site selection entailed asking community informants to identify locations frequented by young bicycle riders. In the bicycle club/map (CLMA) method, site selections were based on recommendations from club members of sites at which cyclists were likely to be found and through examination of maps, keying on local features. These alternative site selection methods were compared in terms of their overall and cost effectiveness in locating youth riders. RESULTS: Despite fewer observer hours and fewer sites in a sparsely populated rural county, the COBA method yielded greater numbers of riding youth and from 1.9 to 4.6 times more youth riders per observer hour than did the CLMA method in two densely populated suburban counties. In addition, costs per youth rider observed associated with the COBA method were 2.9 to 7.0 times lower than those associated with the CLMA method. CONCLUSIONS: Community based site identification is both more efficient in locating youth riders and more cost effective. PMID- 9346111 TI - Emergency Medical Services for Children: it could save a child's life. PMID- 9346110 TI - Demographics of alpine skiing and snowboarding injury: lessons for prevention programs. AB - OBJECTIVE: To establish the demographics of ski injury in relation to age, gender, and perceived cause during a representative season to identify potential injury prevention strategies. SETTING: Blackcomb Mountain, a world class ski resort in British Columbia, Canada. METHODS: Data were collected from the lift ticket records and from ski patrol injury reports for one season, November to May 1991-2. RESULTS: There were 720,066 skier and snowboarder day visits counted by the mountain's lift ticket records, with a total of 2,092 injury reports (incidence 2.91 per 1,000 day visits). Of those with significant injuries (those requiring physician care), 1,210 (58%) were male. The highest injury rate was among children (age 7-12) and teens (age 13-17) with incidences of 3.18 and 3.34 significant injuries per 1,000 skier days, respectively. Head and face injuries constituted 17% and 22% of injuries, respectively in these groups. Overall 22% of head and face injuries were severe enough to cause loss of consciousness or clinical signs of concussion. This was the body region injured most frequently in males. For females over 7 years of age, the knee was the most common site of injury. For youths, the incidence of injuries during school organized activities was 25% higher than during other outings. CONCLUSIONS: The vulnerability of school group participants suggests special education is warranted. The high incidence of head injuries, particularly among young males, needs to be addressed. In light of the high proportion of this group who already wear helmets, the role of helmets in both protection and possible causation of head injury needs objective research. PMID- 9346112 TI - Fractures caused by bicycling. PMID- 9346113 TI - Bicycle helmet use. PMID- 9346139 TI - Scoliosis induced by asymmetric lordosis and rotation: an experimental study. AB - STUDY DESIGN: An experimental study of the influence of intrinsic muscle imbalance on the spinal column of a growing rabbit. OBJECTIVES: To create an in vivo experimental model of scoliosis for comparison with human scoliosis. SUMMARY OF BACKGROUND DATA: There is evidence that asymmetric lordosis may produce scoliosis and that there is muscle imbalance in scoliotic patients. METHODS: Surgical tethering of the spinous apophysis and transverse apophysis of rabbits was performed at three upper levels on the same side of the spine to simulate dominance of one side of the paravertebral musculature over the other. RESULTS: All animals exhibited scoliosis that was convex toward the side opposite that receiving surgery. Radiography showed the curve to increase with time. Postmortem examination of vertebrae revealed structural alterations similar to those produced in human scoliosis. CONCLUSIONS: Intrinsic muscle imbalance in the spinal column of the experimental growing animal may produce scoliosis with characteristics similar to those of human idiopathic scoliosis. PMID- 9346140 TI - Stabilizing function of trunk flexor-extensor muscles around a neutral spine posture. AB - STUDY DESIGN: This study examined the coactivation of trunk flexor and extensor muscles in healthy individuals. The experimental electromyographic data and the theoretical calculations were analyzed in the context of mechanical stability of the lumbar spine. OBJECTIVES: To test a set of hypotheses pertaining to healthy individuals: 1) that the trunk flexor-extensor muscle coactivation is present around a neutral spine posture, 2) that the coactivation is increased when the subject carries a load; and 3) that the coactivation provides the needed mechanical stability to the lumbar spine. SUMMARY OF BACKGROUND DATA: Theoretically, antagonistic trunk muscle coactivation is necessary to provide mechanical stability to the human lumbar spine around its neutral posture. No experimental evidence exists, however, to support this hypothesis. METHODS: Ten individuals executed slow trunk flexion-extension tasks, while six muscles on the right side were monitored with surface electromyography: external oblique, internal oblique, rectus abdominis, multifidus, lumbar erector spinae, and thoracic erector spinae. Simple, but realistic, calculations of spine stability also were performed and compared with experimental results. RESULTS: Average antagonistic flexor-extensor muscle coactivation levels around the neutral spine posture as detected with electromyography were 1.7 +/- 0.8% of maximum voluntary contraction for no external load trials and 2.9 +/- 1.4% of maximum voluntary contraction for the trials with added 32-kg mass to the torso. The inverted pendulum model based on static moment equilibrium criteria predicted no antagonistic coactivation. The same model based on the mechanical stability criteria predicted 1.0% of maximum voluntary contraction coactivation of flexors and extensors with zero load and 3.1% of maximum voluntary contraction with a 32 kg mass. The stability model also was run with zero passive spine stiffness to simulate an injury. Under such conditions, the model predicted 3.4% and 5.5% of maximum voluntary contraction of antagonistic muscle coactivation for no extra load and the added 32 kg, respectively. CONCLUSIONS: This study demonstrated that antagonistic trunk flexor-extensor muscle coactivation was present around the neutral spine posture in healthy individuals. This coactivation increased with added mass to the torso. Using a biomechanical model, the coactivation was explained entirely on the basis of the need for the neuromuscular system to provide the mechanical stability to the lumbar spine. PMID- 9346141 TI - Axial rotation strength in seated neutral and prerotated postures of young adults. AB - STUDY DESIGN: To determine the trunk-twisting capability from neutral and prerotated postures, a study was designed to measure torque generated in isometric and isokinetic activities of 50 young adults. OBJECTIVES: To determine the isometric and isokinetic axial rotation strengths of male and female subjects in neutral and asymmetric postures and to quantify the effect of velocity of rotation on the isokinetic trunk strength profile. METHODS: A specially designed axial rotation tester was employed using specially written modular software for data collection and analysis. The isometric strengths were measured in neutral, 15 degrees, and 30 degrees prerotated trunk postures. The isokinetic strength was measured in activities starting from the neutral position to fully rotated and from a fully rotated position to neutral positions at 10 degrees, 20 degrees, and 40 degrees per second angular velocity. The data obtained were subjected to multivariate and univariate analyses of variances with multiple comparisons and multiple regression analyses. RESULTS: All study participants were significantly stronger in isometric twisting activities than in the isokinetic activities. In isometric activities, participants were 20-25% weaker in prerotated postures when twisting in the direction of prerotation and were approximately 30% stronger in the opposite direction. For isokinetic activities, the trunk rotation from neutral to asymmetric positions produced lesser torques compared with torques from rotated positions to the neutral position. The torque-producing capability declined with increasing velocity of activity. CONCLUSION: This study adds to the data base of rotational strength. PMID- 9346142 TI - Analysis of harvest morbidity and radiographic outcome using autograft for anterior cervical fusion. AB - STUDY DESIGN: Retrospective study of 184 autologous iliac crest bone grafts used for anterior cervical fusion in 144 procedures. OBJECTIVES: To evaluate the effect of autologous iliac crest bone graft harvest site on operation and recovery and to identify patients at risk for harvest morbidity. SUMMARY OF BACKGROUND DATA: Although autologous iliac crest bone graft is considered the most successful grafting material, concerns about harvest morbidity provide a rationale for considering allograft. Data about the use of autograft therefore would assist spinal surgeons in selecting the appropriate substrates for fusion after anterior cervical decompression. METHODS: Statistical analysis based on patient gender, smoking history, obesity, and medical or pharmacologic risk factors for wound healing was used to evaluate morbidity after patient interviews and examinations. Limited assessment of radiographic outcome also was performed. RESULTS: A second operation because of donor site morbidity was performed in four patients (2.8%), but only one (0.7%) with meralgia paresthetica had permanent sequelae. Superficial wound infection or dehiscence occurred in 5.6% of patients, with a disproportionate number of women, obese patients, and those with medical risk represented. Protracted wound symptoms of pain and poor cosmesis were reported in 2.8% and 3.5% of patients, respectively, and also were found in a significant number of female and obese patients. Evidence of fusion was present in 97% of cases. CONCLUSION: Autologous iliac crest bone graft harvest results in minimal major morbidity when regional anatomy is respected and careful technique is observed. The identification of patients at risk for minor complications suggests that allograft may be appropriate in these patients; however, prospective comparison is required to identify whether graft material or technical factors determine fusion success and relative benefit. PMID- 9346143 TI - Scoliosis and developmental theory: adolescent idiopathic scoliosis. AB - STUDY DESIGN: Statistical analysis of maturity and asymmetry criteria in adolescent idiopathic scoliosis. OBJECTIVES: To explore the hypothesis that scoliosis is a manifestation of developmental destabilization under physiologic stress. BACKGROUND: Advances in genetics and theoretical biology have broadened the understanding of morphogenesis and the controlling mechanisms for the development of the adult form. The morphologic genome can be viewed as a cybernetic control system, and the homeobox genes can be viewed as the master controls for the specific subroutines. Deformity will occur from a faulty "program" or a disturbance in the running of that program. An early and subtle indication of such an occurrence is failure of bilateral symmetry. METHODS: An analysis of variance in Cobb angle, age at diagnosis, and apical site in 327 girls with spinal curves 5 degrees or greater according to prospectively maintained scoliosis screening records was performed. Dermatoglyphics were compared in 114 female control individuals and 164 female patients with adolescent idiopathic scoliosis (minimum Cobb angle, 10 degrees), and then by subdivision into school screening (n = 86) and general clinic referrals (n = 78). RESULTS: Girls from the screening program (from the years 1979-1990) had statistically significant associations between age and apical vertebra, suggesting an age-deformity relationship independent of maturation and growth spurt. Girls with adolescent idiopathic scoliosis from a later cohort, both screened and unscreened, had increased directional asymmetry, as previously reported. There was a statistically significant increase in fluctuating asymmetry in general clinic referrals when compared with school screening referrals. CONCLUSIONS: Developmental instability can explain adolescent idiopathic scoliosis as part of wider developmental theory without the necessity of a disease process in the etiology. The differences between screening and general clinic referrals suggest the need for natural history studies, and treatment protocols also should consider the provenance of the individuals described. PMID- 9346144 TI - Pedicle screw instrumentation of the thoracic spine in idiopathic scoliosis. AB - STUDY DESIGN: A prospective study of the accuracy of thoracic pedicle screw placement in patients with idiopathic scoliosis. OBJECTIVES: To evaluate the accuracy of thoracic pedicle screw placement in the surgical management of idiopathic scoliosis and to establish its risks and benefits. SUMMARY OF BACKGROUND DATA: Lumbar pedicle screw instrumentation has proven to be reliable and effective in the surgical management of scoliosis. No reports exist on the accuracy and benefits of pedicle screw instrumentation of the thoracic spine in scoliosis surgery. METHODS: One hundred and twenty thoracic pedicle screws in 32 consecutively treated patients with idiopathic scoliosis were investigated immediately after surgery by computed tomography scans that were analyzed by three examiners. RESULTS: Thirty (25%) of the screws penetrated the pedicle cortex or the vertebral body anterior cortex. Ten screws (8.3%) penetrated the medial cortex of the pedicle by an average of 1.5 mm and a maximum of 3.0 mm. Seventeen screws (14.2%) penetrated laterally by an average of 2.1 mm. There were two cases of caudad penetration. Three screws penetrated the anterior vertebral cortex, of which two also penetrated the pedicle cortex. Also, one of these three screws was replaced because of its direct proximity to the thoracic aorta. There were no neurologic complications. The correlation between the pedicle cortical penetration rate and the preoperative Cobb angle, vertebral rotation or level, or site of screw insertion was statistically insignificant (P > 0.05). Curve correction in the cases of mainly hook instrumentation averaged 52.5% versus 59.2% in the cases of mainly screw instrumentation. This difference was statistically insignificant (P > 0.05). CONCLUSIONS: Pedicle or vertebral body cortical penetration occurred with 25% of the screws but with no neurologic compromise. Curve correction was slightly greater than with hooks, but not to a statistically significant extent. PMID- 9346146 TI - The effects of comorbidity and other factors on medical versus chiropractic care for back problems. AB - STUDY DESIGN: This study is a cross-sectional analysis of adults in the United States who reported at least one back-related visit to a health care professional during a 2-week reference period. OBJECTIVES: To estimate and compare the effects of comorbidity and other factors on self-reported use of medical and chiropractic care for back problems in the United States. SUMMARY OF BACKGROUND DATA: Although back pain is the second most frequent primary symptom reported by patients seeking medical care and the most frequent primary symptom among chiropractic patients, there is a dearth of research on the predictors of chiropractic and medical care among back pain patients. METHODS: Data from the 1989 National Health Interview Survey were used to perform a cross-sectional analysis of adults who sought care for a back-related condition. The primary predictor variables included comorbidity and associated disability, sociodemographic variables, and back-problem-related variables. Weighted logistic regression modeling was performed to estimate odds ratios adjusted for the effects of covariates. RESULTS: Of the 4790 adults with reported back problems, 931 sought health care for their back condition during the 2-week reference period. Adults with disabling comorbidities and back-related restricted-activity days were relatively less likely to use chiropractic care than primary medical care. Those who were male, high-school educated, single, employed, and with more than nine doctor visits during the previous 12 months were relatively more likely to use chiropractic care than primary medical care. CONCLUSIONS: The presence of comorbidity-related or back-related disability, as well as other factors, affect the type of care sought for back conditions among adults in the United States. PMID- 9346145 TI - Chronic low back pain rehabilitation programs: a study of the optimum duration of treatment and a comparison of group and individual therapy. AB - STUDY DESIGN: Eighty-four patients with chronic low back pain were treated using cognitive behavioral principles on a pain management program. Outcome data were collected at four points: 10 weeks before treatment, immediately before and immediately after treatment, and 6 months after treatment. In part 1 of the study, patients were assigned randomly to group or individual treatment contexts. In part 2 of the study, patients were assigned randomly to programs of 15, 30, or 60 hours duration. OBJECTIVES: To identify the differences in outcome between programs that treated patients as part of a group and those that treated patients individually and the effects of duration of treatment on outcome. SUMMARY OF BACKGROUND DATA: Cognitive behavioral programs have been shown to be an effective means of managing chronic low back pain. The literature is concerned with group programs, however, the duration of which vary widely. METHOD: Psychological and functional variables were measured before and after treatment and at the 6-month follow-up visit. Changes in these variables were measured, and comparisons were made between group and individual programs and between 15-, 30-, and 60-hour programs. RESULTS: Data analysis showed a significant, beneficial effect of intervention in terms of the majority of variables; however, these changes were generally independent of whether patients were treated as part of a group or individually and whether patients completed a 15-, 30-, or 60-hour program. CONCLUSIONS: Cognitive behavioral rehabilitation programs have been demonstrated to be an effective means of reducing psychological distress, of changing cognition, and of improving the function of patients with chronic low back pain; however, the length of program and whether patients were treated individually or as part of a group did not affect outcome. This finding has clinical and economic implications. PMID- 9346147 TI - A population-based study of reoperations after back surgery. AB - STUDY DESIGN: Longitudinal follow-up study of back surgery reoperations using an administrative database. OBJECTIVES: To identify population-based rates and factors that determine the need for reoperation after back surgery. SUMMARY OF BACKGROUND DATA: Reoperation after lumbar surgery has poorer results than the initial surgery, yet the population-based incidence and determinants of reoperation are not known. Reported rates of reoperation are derived from retrospective case series and range from 4% to 15%. There are conflicting data on the rate of reoperation after different types of initial surgery. METHODS: All patients who had back surgery in the Province of Ontario (population 10,000,000) between April 1990 and March 1991 were identified using hospital discharge abstracts and an ICD-9 code algorithm. Patients who had undergone prior surgery were excluded. Patients were observed from the index operation to subsequent readmission and reoperation with a maximal time to follow-up examination of 4 years. Basic demographic information and information regarding diagnoses, surgery performed, complications, comorbid factors, reoperation diagnosis, and surgery type were obtained. Patients were divided into surgical treatment groups, and their subsequent reoperations were identified. Multivariate analysis using proportional hazards modeling was conducted. RESULTS: The index surgery group consisted of 4,722 patients, of whom 449 (9.5%) underwent reoperations in the follow-up period. Complications from surgery were significantly higher in the fusion and fusion with decompression groups. The reoperation rate was not significantly different among individual surgery groups. Diagnosis, operation performed, complications after the index surgery, comorbid conditions, and sex did not predict the need for spine reoperation. Younger age was predictive of the likelihood of reoperation (P = 0.04) CONCLUSION: The incidence of reoperation after back surgery is independent of diagnosis and type of surgery performed. Despite different anatomic reasons for surgical intervention, the success of different types of surgery are not influenced by the factors identified in this study. More extensive surgery does not prevent nor predispose a patient to the need for further surgery. PMID- 9346148 TI - The use of intrathecal morphine for analgesia after posterolateral lumbar fusion: a prospective, double-blind, randomized study. AB - STUDY DESIGN: A prospective, randomized, double-blind, placebo-controlled study was undertaken to evaluate the efficacy and safety of intrathecal morphine for postoperative analgesia after posterolateral lumbar fusion. OBJECTIVES: To compare the early postoperative analgesia in patients who receive a single dose of intrathecal morphine intraoperatively with that of patients using a patient controlled analgesia pump only. SUMMARY OF BACKGROUND INFORMATION: Although intrathecal morphine is used as an analgesic in a variety of medical and surgical conditions, very little has been published on its use after posterior lumbar spine surgery. Because the thecal sac is readily available during these procedures, the addition of a single injection of morphine before wound closure can be done with technical ease. If its efficacy and safety can be verified, then it could serve as a useful adjuvant to the postoperative analgesia regimen. METHODS: Sixty-eight consecutive patients undergoing posterolateral lumbar fusion were randomly assigned to two groups. The experimental group was injected intrathecally with morphine 30 minutes before wound closure, and the control group was similarly injected with a placebo of normal saline solution. All patients were connected to an on-demand patient-controlled analgesia pump to provide any additional necessary analgesia. Their use of the patient-controlled analgesia pump was tabulated by counting the number of demands and the actual amount of morphine delivered. Additionally, a visual analog scale was used to assess pain levels at pre-established regular intervals. RESULTS: The visual analog scale measurements were significantly lower for the intrathecal morphine group initially, but they surpassed those of the control group after 24 hours. Likewise, the number of patient-controlled analgesia pump demands and the amount of narcotic delivered initially were significantly lower in the experimental patients, but again reversed after the first postoperative day. The late rebound in pain and patient-controlled analgesia pump use did not reach statistical significance. There were no significant complications related to the analgesia. CONCLUSIONS: Intrathecal morphine can be safe and efficacious as an early postoperative analgesic after lumbar fusion when respiratory monitoring is used. PMID- 9346149 TI - Surgical outcome of 438 patients treated surgically for lumbar spinal stenosis. AB - STUDY DESIGN: A retrospective, follow-up study. OBJECTIVES: To investigate the overall outcome of surgery for lumbar spinal stenosis and to investigate the preoperative factors affecting outcome. SUMMARY OF BACKGROUND DATA: The success rates of surgical intervention for lumbar spinal stenosis vary, and few preoperative factors have been found to be significantly correlated to surgical outcome. METHODS: A total of 438 patients (183 women, 255 men) who underwent decompressive surgery for lumbar spinal stenosis were re-examined and evaluated for outcome 4.3 years after surgery. Outcome was based on subjective disability, which was assessed using the Oswestry low back pain questionnaire. The preoperative data (clinical documentation, length of laminectomy, and radiographs) were collected from patient records that had been stored in the hospital. Preoperative factors affecting outcome were reported. RESULTS: The mean value of the Oswestry disability score of these 438 patients was 34 +/- 18 (women, 36.3 +/- 17; men, 32.3 +/- 18; P < 0.05). Age did not influence general outcome. The proportion of good to excellent outcomes of all 438 patients was 62% (women, 57%; men, 65%). Diabetes, hip joint arthrosis, and preoperative fracture of the lumbar spine seemed to be associated with poor outcome. The ability to work before or after surgery and a history of no prior back surgery were predictive of good outcome. CONCLUSION: The results suggest that clear myelographic stenosis and no prior surgical intervention, no comorbidity of diabetes, no hip joint arthrosis, and no preoperative fracture of the lumbar spine are factors associated with a good outcome in surgical management of lumbar spinal stenosis. PMID- 9346150 TI - Extradural sensory rhizotomy in the management of chronic lumbar radiculopathy: a minimum 2-year follow-up study. AB - STUDY DESIGN: Fifty-one consecutive patients who underwent extradural sensory rhizotomy for chronic radiculopathy after lumbar surgery were reviewed retrospectively. OBJECTIVES: To determine the effectiveness of sensory rhizotomy in the management of chronic radiculopathy in patients selected by extensive imaging techniques and selective nerve root sheath injections. SUMMARY OF BACKGROUND DATA: Results of more central ablative procedures for chronic benign pain problems have been disappointing, with variable reports of pain relief. METHODS: Fifty-one patients were reviewed. All patients underwent extensive evaluation to exclude reversible structural lesions, and all had the diagnosis of chronic radiculopathy confirmed by results of clinical and electrophysiologic examination. Selective nerve root sheath injections under fluoroscopic guidance confirmed the symptomatic nature of the segments. All blocks were repeated at least once. All patients underwent selective sensory rhizotomy or, in some cases, complete rhizotomy. After rhizotomy, 37 patients were available to be observed at selected time intervals for a minimum of 2 years. Clinical results were determined by the presence or absence of pain relief (visual analog scale), sensory and motor deficits, narcotic analgesic usage, and the patient's estimation of the effectiveness of the procedure. RESULTS: At 6 months after surgery, all 51 patients and the outcomes of their surgery were available for review. Fifty-five percent of patients rated were believed to have good or excellent outcomes, whereas the remainder had poor or failed outcomes. For the minimum 2-year follow-up period (range, 2-4.2 years), 37 patients were available for review. At final follow-up examination only 19% of the patients maintained good or excellent outcomes. CONCLUSIONS: The results of the rhizotomy procedures deteriorated over time. Possible reasons for the failure, other than temporal deterioration, were anatomic factors and lack of specificity of diagnostic techniques, specifically selective nerve root sheath injection. At this point rhizotomy cannot be recommended with any confidence whatsoever in the setting of chronic lumbar radiculopathy after lumbar surgery. PMID- 9346152 TI - The effect of intraoperative hip position on maintenance of lumbar lordosis: a radiographic study of anesthetized patients and unanesthetized volunteers on the Wilson frame. AB - STUDY DESIGN: The effect of intraoperative hip position on maintenance of lumbar lordosis was evaluated radiographically in 13 anesthetized patients and 14 unanesthetized volunteers positioned on a Wilson frame (MDT Corp., Torrance, CA). OBJECTIVES: To evaluate the effect of hip position on total and segmental lumbar lordosis in patients and volunteers in standardized positions: standing and with hips extended and flexed on a Wilson frame. SUMMARY OF BACKGROUND: Preservation of lordosis during instrumented lumbar fusion is critical for maintenance of normal sagittal alignment. It is customary to extend the hips on certain positioning devices to maximize lordosis maintenance. However, little information exists concerning the degree to which this actually affects lumbar lordosis. Further, the question of how individuals are specifically affected intraoperatively by differences of position on the same device remains unanswered. METHODS: Preoperative standing and intraoperative lateral lumbar spine radiographs with patients' hips in standardized flexed and extended positions were obtained (n = 13). Similar radiographs were obtained of asymptomatic volunteers (n = 14). Lumbar lordosis (L1-S1) and intervertebral body angles at each level were measured. Data were analyzed for changes in total and segmental lordosis between standing and intraoperative positions for all subjects. RESULTS: In the patient group, 95% of preoperative standing lordosis was maintained with the patients' hips extended. With hips flexed from 19 degrees to 48 degrees (mean, 33 degrees), 74% of lordosis was maintained. In the volunteer group, 98% of standing lordosis was maintained with volunteers' hips extended; with their hips flexed 20 degrees to 36 degrees (mean, 28 degrees), 86% of lordosis was maintained. CONCLUSIONS: Hip flexion was associated with a significant decrease in lordosis in patients and volunteers. Positioning in maximal hip extension optimizes lordosis preservation. While other devices have been shown to have specific effects on lordosis, the Wilson frame can permit easy adjustment of the lumbar sagittal contour to facilitate either preservation or reduction in lordosis. PMID- 9346151 TI - Vertebral metastases: a critical appreciation of the preoperative prognostic tokuhashi score in a series of 71 cases. AB - STUDY DESIGN: The utility of the Tokuhashi score was assessed in a retrospective study in 71 patients with vertebral metastases. OBJECTIVES: To study the importance of the site of the primary tumor as a parameter in the preoperative prognostic Tokuhashi score. SUMMARY OF BACKGROUND DATA: A preoperative score composed of six parameters, each rated from zero to two, has been proposed by Tokuhashi for the prognostic assessment of patients with metastases to the spine. METHODS: Seventy-one patients with vertebral metastases were studied. There were 34 cases of thyroid cancer metastases, 28 cases of renal cancer metastases, and nine cases of metastases of unknown origin. In each patient, a local and a systemic tumor search were performed. Patients were divided into groups based on the primary site of the tumor, and each group was analyzed separately. RESULTS: In cases of vertebral metastases of thyroid cancers, surgery to excise single metastases was found to provide good results, as was palliative surgery of multiple metastases. Vertebral metastases of renal tumors were rarely single, and the results of palliative surgery were less satisfactory. Vertebral metastases of unknown primary tumors had a poor outcome, regardless of whether surgery was excisional or palliative. The median survival period in patients with metastases of unknown primary tumors was significantly shorter than that in patients with renal or thyroid cancer metastases. CONCLUSION: The Tokuhashi preoperative score is successful as a prognostic tool. However, it attributes the same one-point rating to metastases of renal cancer and to those of unknown primary tumors. In the case of metastases of unknown primary tumors, this rating is too high and should be reduced to 0. PMID- 9346153 TI - Subacute paraparesis induced by venous thrombosis of a spinal angiolipoma: a case report. AB - STUDY DESIGN: A case report of spinal extradural angiolipoma, a rare tumor that can cause spinal cord compression, is presented with a complete review of the literature related to this disorder. OBJECTIVES: To discuss venous thrombosis involving the angiolipoma in the development of subacute paraparesis. SUMMARY OF BACKGROUND DATA: This case shows that venous thrombosis of a spinal angiolipoma can precipitate the subacute onset of paraparesis. METHODS: Medical history, physical findings, and the results of imaging and histopathologic studies were analyzed to elucidate the pathogenesis of the patient's subacute onset of paraparesis. A bilateral T3-T7 laminectomy was performed, and although the tumor was extremely hemorrhagic, it was mobilized easily off the compressed dura to achieve resection. RESULTS: The postoperative course was uneventful. One month after her surgery, the patient's myelopathic symptoms had resolved, and the she was able to return to work. CONCLUSION: Because the prognosis after surgical management of these lesions is favorable, the diagnosis of thrombosis involving a spinal angiolipoma should be considered in the differential diagnosis of subacute spinal cord compression. PMID- 9346155 TI - Spine epidural steroids for patients with lumbar spinal stenosis. AB - The use of epidural steroid injections to relieve sciatic pain from spinal stenosis is extremely variable and controversial. Drs. Cohen and Kostuik take the position that most studies do not support their use and highlight the potential complications. Dr. Rydevik believes that epidural steroids might be considered as a nonsurgical alternative, especially in elderly patients where surgery carries greater risk. PMID- 9346154 TI - Gouty arthropathy of the lumbar spine: a case report and review of the literature. AB - STUDY DESIGN: A patient with hyperuricemia developed symptoms from lateral recess stenosis attributed to gouty arthropathy of a lumbar facet joint. OBJECTIVE: To present the diagnosis and management of gouty arthropathy of the lumbar spine in one individual. SUMMARY OF BACKGROUND DATA: The symptoms and treatment of a patient with intra-articular gout of a lumbar facet are presented and contrasted with other cases of spinal extra-articular gout found in the literature. METHODS: A patient with hyperuricemia reported back pain and symptoms consistent with lateral recess stenosis. Conservative treatment failed, and, after further evaluation, a successful decompressive laminectomy was performed. Pathology revealed intra-articular urate crystal deposition. RESULTS: This patient's unilateral S1 radiculopathy corresponded with magnetic resonance and computed tomography studies documenting unilateral lateral L5-S1 lateral recess stenosis secondary to intra-articular gouty arthropathy. As anticipated, the serum uric acid also was elevated. Since surgical decompression with unilateral laminotomy was performed, the patient has been symptom-free for 2 years. CONCLUSION: Although rare, gouty arthropathy of the lumbar facet joint should be considered in all patients with neurologic symptoms and known or suspected gout. Optimization of pharmacologic treatment is indicated for patients suspected of having gouty neuropathy. Surgical decompression is indicated if conservative management with Indocin, nonsteroidal anti-inflammatory agents, and allopurinol fails to reverse neurologic dysfunction. PMID- 9346156 TI - Predictive validity of an injury score among high school basketball players. AB - A number of strategies have been investigated in an attempt to identify those individuals most likely to be injured during participation in sports activity. One such strategy identified in the literature involved computing an "injury score" via a logistic regression equation using measures of structural symmetry to predict the likelihood of athletic injury. The purpose of this study was to investigate the predictive value of the injury score among high school basketball players. Following the establishment of reliability of measures, injury scores were calculated for 62 high school basketball players (34 females, 28 males) before the start of the season. Lower extremity injuries sustained while playing basketball were recorded throughout the season. The predictive value of the injury score equation was determined by calculating sensitivity, specificity, and positive and negative predictive values. The sensitivity and specificity were calculated to be 16.7% and 66.1%, respectively. The positive and negative predictive values were calculated to be 5.90% and 88.1%, respectively. These results indicate that the injury prediction score investigated was not a valid means of predicting injury in high school basketball players. Limitations, possible implications of these findings, and ideas for future related research are presented. PMID- 9346157 TI - Improved walking economy in patients with peripheral arterial occlusive disease. AB - The effect of exercise rehabilitation on the oxygen cost of ambulation in patients with peripheral arterial occlusive disease (PAOD) was evaluated with specific emphasis on the effects of exercise rehabilitation on the slow component of VO2. Because the slow component of VO2 represents an increase in VO2 despite constant-intensity exercise, it can profoundly affect the relative energy cost of exercise in individuals with a low functional capacity. Twenty-six patients with intermittent claudication performed treadmill walking at 2.0 mph/0% grade for 20 min or until maximal claudication pain before and after 4 months of rehabilitation. The slow component of VO2 during the treadmill test was defined as the difference between the end-exercise VO2 and the VO2 observed at minute 3. Ankle/brachial systolic pressure index (ABI) was measured before and immediately following the exercise test. Rehabilitation consisted of 3 d x wk(-1) of treadmill walking for 15-30 min at 60-70% of VO2peak. The slow component of VO2 and end-exercise VO2 at pretraining (0.75 +/- 0.90 and 11.12 +/- 2.10 mL x kg[-1] x min[-1]) were significantly reduced after 4 months of exercise rehabilitation ( 0.07 +/- 1.11 and 10.07 +/- 1.80 mL x kg[-1] x min[-1]; P < 0.05). Exercise rehabilitation also significantly (P < 0.05) increased the post-exercise ABI (pre rehabilitation = 0.36 +/- 0.26, post-rehabilitation = 0.43 +/- 0.25). These data suggest that 4 months of exercise rehabilitation: 1) improves walking economy in PAOD patients because of a decreased slow component of VO2, and 2) increases post exercise ABI. PMID- 9346158 TI - Lower extremity alignment and risk of overuse injuries in runners. AB - A group of 304 runners enrolling in a marathon training program had alignment measurements performed and completed a questionnaire on training practices and injuries over the previous 12 months. The alignment measures consisted of arch index (AI), heel valgus (HV), knee tubercle-sulcus angle (TSA), knee varus (KV), and leg-length difference (LLD). Results indicated few consistent statistical associations between these alignment measures and risk of injuries, either bivariately or multivariately: left AI with hamstring injuries; right AI with shin injuries; right HV with back injuries; left TSA with ankle injuries; KV with hip injuries; and LLD with back, ankle, and foot injuries. A few statistically significant relationships were also found between other training and anthropometric factors and injuries: mileage with hamstring injuries; interval training with shin injuries; hard surfaces with back and thigh injuries; shoe use patterns with foot and overall injuries; and body mass index with heel injuries. We conclude that lower-extremity alignment is not a major risk factor for running injuries in our relatively low mileage cohort; however, prospective studies are necessary to confirm or refute these findings. PMID- 9346159 TI - Cardiovascular response to sudden strenuous exercise: an exercise echocardiographic study. AB - We studied the response in left ventricular (LV) internal dimensions and posterior wall thickness during the performance of sudden strenuous exercise without warm-up (SSE) and sudden strenuous exercise with warm-up (SSEw) in 15 healthy, untrained college-aged males (26 +/- 5.0 yr). Measurements of left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic dimension (LVESD), stroke dimension (SD = LVEDD-LVESD), fractional shortening (FS = SD/EDD) and posterior wall thickness (PWT) were obtained from continuous 2-D targeted on M-mode echocardiography. Continuous EKG and blood pressure were obtained at rest and during the final 10 s of SSE (30 s of upright leg cycle ergometry of 400 W at 80 rpm). SSEw was preceded by warm-up exercise (6 min of graded leg cycle exercise of 2-min stages initial load 30 W, increasing in 30-W increments at 60 rpm, followed immediately by SSE). Our findings revealed that there were no significant differences in the LV internal dimensions (LVEDD, LVESD, FS, PWT), HR max, RPP max, ECG, and MAP max between SSE and SSEw. Sudden strenuous exercise without warm-up is not associated with a reduced LV function. The results of this study are contradictory to previous findings that have suggested that SSE is associated with transient global left ventricular (LV) dysfunction. PMID- 9346160 TI - Arterial morphology and blood volumes of rats following 10-14 weeks of tail suspension. AB - We hypothesized that the structure of systemic arteries would be altered following 10-14 wk of hindlimb unloading (tail suspension) in female Sprague Dawley rats. Tail suspension resulted in atrophy of the soleus muscle (P < or = 0.01) but no significant differences in the mass of the extensor digitorum muscle, heart, or adrenal glands. In anesthetized rats, there was no difference between groups in arterial pressure (approximately 60 mm Hg). The corresponding maximal (topical papaverine) external diameter (ED) of femoral arteries (N = 5 per group) was reduced (P < or = 0.05) in tail suspended (TS, 511 +/- 47 microm, mean +/- SD) compared with cage sedentary (CS, 615 +/- 89 microm) and food restricted weight-paired (FR, 643 +/- 61 microm) groups. Neither hematocrit, red cell, plasma, nor total blood volume differed among groups. Following systemic vasodilation with papaverine, progressive arterial inflation with liquid silicon rubber (Microfil) revealed a reduction in both ED and distensibility of the femoral artery (P < or = 0.05). To determine the effects of tail suspension on systemic arterial morphology, the vasculature of additional rats was perfusion fixed at 80 mm Hg during vasodilation. Cross sections (thickness, 8 microm) of the carotid, axillary, iliac, and femoral arteries were then evaluated. Whereas the internal diameter of femoral arteries was smaller in TS than in CS (P < 0.05), no differences were observed for other vessels among groups. Further, arterial wall thickness increased systemically (overall, P < 0.05; carotid, 24%, P < 0.01; femoral, 28%, P < 0.01) following tail suspension. These findings illustrate adaptation in the structure of conduit arteries to prolonged tail suspension, with diameter altered regionally and wall thickness increased systemically. We suggest that chronic changes in activity patterns can influence arterial structure. PMID- 9346161 TI - Muscle-specific creatine kinase gene polymorphism and VO2max in the HERITAGE Family Study. AB - This study examined the association between a DNA polymorphism in the muscle specific creatine kinase (CKMM) gene and VO2max in the sedentary state, as well as its response (deltaVO2max) to a standardized 20-wk endurance training program. The subjects were 160 biologically unrelated Caucasian parents (80 women, 80 men) and 80 biologically unrelated adult offspring of the HERITAGE Family Study. The CKMM polymorphism was detected by PCR and digestion with the NcoI restriction enzyme. VO2max was measured during maximal cycle ergometer tests. VO2max was 2119 +/- 45 mL x min(-1) (mean +/- SE) or 26 +/- 0.4 mL x kg(-1) x min(-1). Both sexes had a significant (P < 0.05) increase in the deltaVO2max (women = 283 +/- 20 mL x min[-1] and men = 363 +/- 25 mL x min[-1]). Allele and genotype frequencies were not significantly different (P > 0.05) between sexes. Age and sex adjusted VO2max was significantly (P = 0.007) associated with the CKMM genotype in the parents, whereas no association (P > 0.05) was observed in the offspring. DeltaVO2max values adjusted for age, sex, VO2max, and body mass were characterized by genotype differences in both parents (P = 0.0004) and offspring (P = 0.0025). A significantly (P < 0.05) lower deltaVO2max to endurance training was detected in both parents and offspring homozygotes for the rare allele. The genotype accounted for at least 9% of the variance in deltaVO2max. These results indicate that the NcoI polymorphism in the 3' untranslated region of the muscle-specific creatine kinase gene is associated with the deltaVO2max to endurance training. PMID- 9346162 TI - Carbohydrate affects natural killer cell redistribution but not activity after running. AB - This randomized, double-blind, placebo-controlled study was designed to determine the influence of carbohydrate supplementation on the natural killer cell response to 2.5 h of high-intensity running (76.7 +/- 0.4% VO2max). Thirty experienced marathon runners (VO2max 53.4 +/- 1.0 mL x kg[-1] x min[-1], age 41.5 +/- 1.4 yr) were randomized into carbohydrate supplement (N = 17) and placebo (N = 13) groups. Subjects rested for 10-15 min before a blood sample at 0715, and then ingested 0.75 L of carbohydrate beverage (Gatorade) or placebo. At 0730, subjects began running at 75-80% VO2max for 2.5 h and drank 0.25 L of carbohydrate or placebo fluid every 15 min. Immediately after the 2.5 h run (1000), another blood sample was taken, followed by 1.5 h, 3 h, and 6-h recovery samples. Carbohydrate supplementation versus placebo had a significant effect on the pattern of change in glucose, cortisol, and the blood concentration of natural killer cells ([F (4,25) = 3.79, P = 0.015], but not natural killer cell activity following 2.5 h of intensive running. PMID- 9346164 TI - Blood lactate and perceived exertion relative to ventilatory threshold: boys versus men. AB - The purpose of this study was to examine blood lactate (BLa) levels and ratings of perceived exertion (RPE) in nine boys (10.5 +/- 0.7 yr) and nine men (25.3 +/- 2.0 yr) during exercise relative to ventilatory threshold (VT). VT and VO2max were determined during a graded exercise test on a cycle ergometer. On three additional days each subject exercised for 10 min at either 80, 100, or 120% of the VO2 at VT. Capillary BLa levels and RPE were assessed at minutes 5 and 10 of each trial. VO2max averaged 47.7 +/- 5.4 and 50.2 +/- 6.2 mL x g(-1) x min(-1) in the boys and men, respectively (P > 0.05). VT expressed as %VO2max was 67.2 +/- 3.5% in the boys and 67.3 +/- 4.9% in the men (P > 0.05). BLa levels ranged from 2.0 +/- 0.7 to 4.7 +/- 0.9 mmol x L(-1) in the boys and from 2.6 +/- 0.5 to 8.2 +/- 2.1 mmol x L(-1) in the men across the three intensities. Corresponding RPE values ranged from 11.2 +/- 1.8 to 16.2 +/- 2.2 in the boys and from 10.2 +/- 1.2 to 15.8 +/- 1.7 in the men. A group x time x intensity interaction (P < 0.05) indicated that BLa in the men increased more so across time and intensity. There were no significant group difference or interactions involving RPE during exercise. Setting exercise intensity relative to VT did not abolish child-adult differences with respect to submaximal BLa levels. Despite maintaining lower BLa levels, RPE values were similar between boys and men. PMID- 9346163 TI - Effect of casting on forearm resistance vessels in young men. AB - The aim of this study was to determine whether physical deconditioning, induced by 6 wk of forearm casting, and the conditioning effects of recovery from casting would elicit changes in the response of forearm resistance vessels to substances that modulate vascular resistance. Forearm blood flow (FBF) responses to intrabrachial infusion of NGmonomethyl-L-arginine (LNMMA) and norepinephrine (NE) were examined in six subjects recovering from Colles', scaphoid, or metacarpal fractures within 72 h of cast removal and again after a 6-wk recovery period. Vascular responses were also examined in six noncasted controls. The FBF responses of casted and control subjects did not differ, and no changes occurred over the 6-wk study period. These results suggest that the effect of forearm casting and recovery from casting do not greatly influence control of vascular tone, including the basal activity of the NO dilator system in vivo. PMID- 9346165 TI - Exercise effect on canine and miniswine cardiac catecholamines and enkephalins. AB - Chronic exercise changes cardiac function and responsiveness to autonomic control. Catecholamines and enkephalins are neuroendocrine transmitters involved in autonomic regulation and signaling. We hypothesized that intermittent increased sympathetic stimulation caused by exercise training would decrease cardiac catecholamine and enkephalin content. Dogs and miniswine were exercise trained and hearts extracted for catecholamine and enkephalin measurements. Atrial catecholamine content is greater than ventricular content in dog heart which is in keeping with the greater atrial neuronal density. In contrast, porcine epinephrine content was evenly distributed across heart sections and norepinephrine content was greater on the right side than the left. Changes in miniswine and dog heart catecholamine content after exercise training were different. Canine cardiac norepinephrine content decreased and porcine norepinephrine content increased. This indicates a difference in cardiac adrenergic control in miniswine versus canines. Methionine-enkephalin (met-enk) distribution across canine heart is uniform, unlike the miniswine, where the atria contain more than the ventricles. Proenkephalin processing produces four met-enk sequences and one met-enk-arg-phe; despite this, ventricles of both species contain more met-enk-arg-phe immunoreactivity than met-enk. Therefore, proenkephalin processing is incomplete in heart tissue. Exercise training in the dog resulted in decreased cardiac met-enk, decreased left atrial met-enk-arg-phe, and increased ventricular met-enk-arg-phe. Porcine cardiac enkephalin concentration was unchanged by training. The changes in the enkephalins may be explained by changes in proenkephalin processing and/or release. Met-enk-arg-phe is particularly good at modulating vagal stimulation of the canine heart. The changes in tissue content seen after exercise training may be a result of the exercise-induced change in autonomic tone to the heart. These data suggest species dependent changes in autonomic regulation. PMID- 9346166 TI - Measuring general levels of physical activity: preliminary evidence for the Physical Activity Questionnaire for Older Children. AB - This article reports three studies that investigated psychometric properties of the Physical Activity Questionnaire for Older Children (PAQ-C). The PAQ-C is a guided self-administered 7-day recall measure designed to assess general physical activity levels during the school year for children in grades four and higher. Study one, with 215 students ranging in age from 9 to 15 yr, found the PAQ-C had acceptable item and test score characteristics such as item distribution, corrected item-total correlations, and internal consistency. Study two, involving 84 students ranging from 9 to 14 yr, indicated acceptable levels of test-retest reliability for both males (r = 0.75) and females (r = 0.82) after 1 wk. The third study used Generalizability theory to investigate the reliability for using the average of either two or three PAQ-C scores collected during fall, winter, and spring seasons. Based on the responses of 200 students ranging from 8 to 16 yr, generalizability coefficients exceeded 0.80 for either the average of two or three responses for both younger (<13 yr) and older subjects. In all three studies, the PAQ-C demonstrated acceptable internal consistency and males were significantly more active than females. These results provide preliminary support for the PAQ-C as a cost efficient method of assessing general levels of children's physical activity during the school year. PMID- 9346167 TI - Lifetime exercise and disk degeneration: an MRI study of monozygotic twins. AB - Participation in some competitive sports has been shown to increase disk degeneration; however, the long-term effects of recreational physical activities are unclear. We investigated the effects of endurance exercise and power sports on disk degeneration in monozygotic male twins with contrasting lifetime exercise histories. The effects of endurance exercise were studied in 22 discordant twin pairs (mean lifetime frequencies of 3.9 vs 1.1 times/wk), and the effects of power sports were investigated in 12 discordant pairs (2,300 vs 200 h of weightlifting). The age range of the twins was from 35 to 69 yr. No differences in MRI findings between co-twins discordant for endurance exercise were found at any of the spinal regions. Subjects with more power sport involvement had greater disk degeneration in the T6-T12 region (P < 0.03), but similar findings were not present in the lumbar spine. Controlling for recalled back injuries, occupational loading, smoking, and driving did not significantly affect the results. No signs of beneficial or harmful effects of lifetime endurance exercise on disk degeneration were seen. Increased power sport participation was associated with slightly greater disk degeneration in the lower thoracic spine, but not in the lumbar spine. PMID- 9346168 TI - Aerobic capacity and cognitive performance in a cross-sectional aging study. AB - In a population unselected for aerobic fitness status, aerobic fitness (VO2max) and its interaction with age were used to predict performance on several cognitive measures known to be affected by chronological age. It was hypothesized that, in particular, cognitively demanding tasks would be sensitive to aerobic capacity. Healthy subjects between 24 and 76 yr of age (N = 132) were recruited from a larger study into determinants of cognitive aging (Maastricht Aging Study MAAS). All participants took part in a submaximal bicycle ergometer protocol and an extensive neurocognitive examination, including tests of intelligence, verbal memory, and simple and complex cognitive speed. Participants engaged more hours a week in aerobic sports and felt healthier than the nonparticipants of the same age did. No group differences were found in the basic anthropometric characteristics height, weight, and BMI. Two of four subtasks that reflect complex cognitive speed (Stroop color/word interference and Concept Shifting Test) showed main and interaction effects with age of aerobic capacity in a hierarchical regression analysis, accounting for up to 5% of variance in parameter score after correction for age, sex, and intelligence main effects. These findings fit well within a moderator model of aerobic fitness in cognitive aging. They add to the notion that aerobic fitness may selectively and age dependently act on cognitive processes, in particular those that require relatively large attentional resources. PMID- 9346169 TI - Ballistic movement performance in karate athletes. AB - Nine male karate athletes and 13 untrained men did maximal voluntary isometric (MVC) and ballistic elbow extension actions, the latter unloaded (L0) and against a load equal to 10% MVC (L10). The karate group achieved greater (P < 0.05) isometric (32%) and ballistic action peak torque with L0 (30%) and L10 (40%). With L10 the ratio of ballistic action to isometric action, peak torque was 13% greater in the karate group, indicating a load specific training adaptation. With L0 the corresponding ratio did not differ significantly between groups. Ballistic action peak rate of torque development (51%, 51%) and peak acceleration (15%, 9%) with L0 and L10, respectively, were greater in the karate group. In contrast, peak velocity and movement time did not differ significantly between groups. Electromyographic recordings of agonist triceps and antagonist biceps were made during the isometric and ballistic actions. Since ballistic actions (L10) were initiated from a preloaded condition, the occurrence and duration of premovement agonist depression were monitored. In ballistic actions there were no group differences in agonist activation, the ratio of ballistic to isometric action agonist activation, or antagonist coactivation. Premovement agonist depression occurred infrequently in both groups, with no group differences. It is concluded that karate athletes have enhanced elbow extension ballistic performance, but it could not be related to amplified agonist activation, altered antagonist activation, or more frequent occurrence of agonist premovement depression. PMID- 9346170 TI - Water running with and without a flotation vest in competitive and recreational runners. AB - The purpose of this study was to determine whether competitive and recreational runners would replicate land training intensity during water immersion (WI) running with (V) and without (NV) a flotation vest and during treadmill running (Tm). Seven female competitive runners (CR) and seven female noncompetitive runners (NR) were asked to replicate preferred land training intensity characteristic of a 45-min run under three conditions (Tm, V, and NV). When 20 min submaximal runs at the preferred land training intensity were performed for Tm, V, and NV conditions, CR were able to elicit a similar submaximal VO2 for all three conditions. In contrast, the NR group had a significantly (P < 0.05) lower VO2 (27%), HR (23%), VE (26%) and %VO2max (27%) during V versus Tm condition. During the NV condition, NR had a significantly lower VO2 (13%), %VO2max (13%), and a higher RPE compared with Tm running, and a significantly higher VO2 (16%), HR (15%), VE (24%), %VO2max (15%) and RPE compared with the V condition. Competitive runners were able to achieve training intensities similar to land training for WI running with or without a flotation vest. However, recreational runners failed to replicate land training pace, where intensity was significantly lower during WI running without a vest and lowest with a vest, despite efforts to maintain a similar level of exertion. PMID- 9346172 TI - Reproducibility of ballistic movement. AB - The reproducibility of ballistic actions performed on a custom-made apparatus was assessed in six untrained men who reported to the laboratory two times over a span of 10 d, on each occasion performing three isometric maximal voluntary contractions (MVC, elbow extension) and 10 ballistic elbow extension movements in each of two conditions: unloaded (L0) and with a load equal to 10% MVC (L10). For both the average of the trials and the best trial, method errors and Pearson correlation coefficients (r) were calculated for isometric peak torque and 10 (5 at each of L0 and L10) selected ballistic movement parameters (peak torque, peak rate of torque development, movement time, peak velocity, and peak acceleration). Based on method errors obtained and given an alpha level of 0.05 and power of 0.9, needed sample sizes were calculated for treatment effects ranging from 5 to 20%. Method errors for isometric peak torque were 5.1 and 3.1% for best and average of trials, respectively. For the 10 ballistic parameters, the corresponding method errors averaged 7.1 +/- 5.4% (SD) and 6.8 +/- 3.6%. The largest method errors were found in peak torque and peak rate of torque development with L0. The expected high negative correlation between method errors and r values was not found. For 5, 10, and 20% treatment effects, 4, 6, and 9 of 10 best trial measures met the sample size criterion of N < or = 20. For the average of trials, 3, 8, and all 10 met the criterion. With the exception of rate of torque development, the reproducibility of the described ballistic measures is acceptable for longitudinal training studies and cross-sectional group comparisons. PMID- 9346171 TI - Acute effects of ski waxing on pulmonary function. AB - The objective of this study was to determine whether pulmonary function is acutely affected by moderate exposure to ski waxing. Ten healthy nonsmoking young adult volunteers were exposed to 45 min of ski waxing in a small unventilated room. The exposure occurred in pairs with one individual performing the waxing while the other overlooked the waxing process. During the period of waxing, two pairs of cross-country skis were waxed with a paraffin wax and then scraped and brushed, and two pairs of cross-country skis were waxed with a fluorinated wax and then brushed. Spirometry and single-breath carbon monoxide lung diffusion capacity (DLCO) were measured immediately before and after exposure to ski waxing, and again 5-6 h after waxing. A subset of five subjects repeated the measurements on a separate day without receiving exposure to ski waxing. Data were analyzed with repeated measures ANOVA. Exposure to ski waxing induced no significant changes in spirometry and DLCO measurements. We conclude that moderate exposure to ski waxing has no significant acute effect on lung function. PMID- 9346173 TI - Swimmers' compliance with training prescription. AB - The purpose of this study was to determine how closely competitive swimmers complied with their coaches' prescriptions when training in squads. A training session early in the buildup phase of the season was observed for each of 24 coaches who had been randomized to two groups: an experimental, high-intensity, low-distance program (E) and a control (usual) program (C). Swim distances, rest durations, and swim durations for at least one set of prescribed repetitions (reps) were recorded for each of 47 swimmers (87 sets, 429 reps) in E and for 49 swimmers (79 sets, 402 reps) in C. The pace of each rep, expressed as percent of the swimmer's current personal best pace for the distance of the rep, represented observed intensity. There was almost perfect agreement between the prescribed and observed swim distances for the set of reps (Spearman r = 0.99 in both groups). Prescribed and observed rest intervals were also closely matched in E and C (Spearman r = 0.87 and 0.77 respectively). Four coaches in E and 10 coaches in C prescribed intensity subjectively as easy, moderate, hard, or race-pace; mean +/- SD observed intensities (%) for their swimmers were 79 +/- 6, 81 +/- 3, 91 +/- 2, and 93 +/- 5, respectively. The relationship between these coaches' subjective training prescription and individual swimmers' interpretations of these intensities was poor (Cohen's kappa = 0.39). Nine coaches in E prescribed intensity as percent of personal best pace. Although the mean prescribed and observed intensities for their swimmers were similar (89 +/- 4 and 90 +/- 7, respectively), the relationship between individual values was poor (Pearson r = 0.30). We conclude that swimmers complied with prescribed distances and rest intervals but were less effective in judging the intensity of swim training. We recommend that coaches monitor training intensity more closely. PMID- 9346174 TI - Risk factors and hormone levels in patients with serous and endometrioid uterine carcinomas. AB - We performed a multi-institutional, incident case-control study of 328 endometrioid and 26 serous carcinomas to assess whether risk factors and circulating hormone levels in women with serous carcinoma differ from the expected profile for endometrial carcinoma We also evaluated exposures potentially related to endometrial cancer risk, anthropometric measurements, and circulating levels of sex hormones and related carrier proteins. Histopathologic specimens were reviewed without knowledge of the other data. As expected, a statistically significant association was observed for high body mass index (BMI) (relative risk, 3.5) and use of menopausal estrogens (relative risk, 2.4) in the endometrioid carcinoma cases, whereas serous carcinomas were not strongly associated with these factors. Smoking and oral contraceptive use decreased risk for both tumor types. For five of six sex hormones tested, age-adjusted mean serum levels in patients with serous carcinoma were significantly lower than those in women with endometrioid carcinoma. After adjustment for BMI, these differences were narrowed, but levels of albumin-bound estradiol and estrone remained significantly lower in the serous cases. Age and BMI-adjusted levels of sex hormone-binding globulin were significantly higher in patients with serous carcinoma than in women with endometrioid carcinomas. In conclusion, risk factors and sex hormone levels in patients with uterine serous carcinoma seem to differ from those in women with endometrioid carcinoma, suggesting that there may be at least two different pathways of endometrial carcinogenesis. PMID- 9346176 TI - Conventional bladder wash cytology performed by four experts versus quantitative image analysis. AB - Bladder wash cytology provides superior results for the detection of bladder malignancies than does voided urine analysis. Image analysis systems have been developed for quantification of cytologic features. In this study, routine bladder wash cytology is compared with an automated image analysis system (QUANTICYT). We studied a random set of 100 bladder wash samples from a population of 1614 patients in follow-up after bladder cancer. Four experienced pathologists interpreted the same 100 Papanicolaou-stained slides. Cytologic and image analysis results were compared for prediction of a cystoscopic lesion, histologic abnormalities, and tumor recurrence. After application of receiver operating characteristic curves, prediction of a cystoscopic lesion by cytology and image analysis was comparable. Both the image analysis system and the cytologic examination detected all of the high-grade lesions. Image analysis was superior to cytologic analysis for the prediction of tumor recurrence after normal findings at cystoscopic examination. PMID- 9346175 TI - Immunohistochemical study of TGF-alpha, TGF-beta1, EGFR, and IGF-1 expression in human breast carcinoma. AB - Localization of growth factors such as transforming growth factor alpha (TGF alpha) and beta1 (TGF-beta1), insulin-like growth factor 1 (IGF-1), and epidermal growth factor receptor (EGFR) in breast cancer tissue is controversial. We immunohistochemically investigated expression patterns of these growth factors and EGFR along with estrogen receptor (ER) status in 36 breast carcinomas (21 invasive ductal, 11 invasive lobular, 4 noninvasive ductal) and compared the results with those found in 10 fibroadenomas. Twenty-four of 36 carcinomas and all of the 10 fibroadenomas showed positivity for ER. TGF-alpha was immunoreactive in all of the carcinomas and fibroadenomas. TGF-beta1 was negative in all of the invasive ductal carcinomas and positive in all of the fibroadenomas and in five lobular carcinomas. EGFR was regularly expressed preferentially in the myoepithelial cells of mammary ducts in the fibroadenomas and in nontumorous glands. Six of the 36 carcinomas were positive for EGFR. Those tumors were negative for ER (P < .001). There was IGF-1 expression in all of the cases of carcinoma and fibroadenoma. We conclude that TGF-alpha is expressed abundantly in invasive and intraductal breast carcinomas and in fibroadenomas. EGFR expression significantly correlates with negative ER status in breast carcinoma. In breast carcinoma, IGF-1 is broadly expressed by the tumor as well as by stromal cells and might act as a growth stimulator in endocrine, paracrine, and autocrine manners. PMID- 9346177 TI - Apoptosis, bcl-2 protein, and Fas antigen in thymic epithelial tumors. AB - We examined 35 thymic epithelial tumors by immunohistochemical techniques to investigate the extent of apoptosis detected by in situ end-labeling of fragmented DNA and the expression of bcl-2 and Fas. There were 22 thymomas (4 medullary, 9 mixed, 3 predominantly cortical, 6 cortical), 3 well-differentiated thymic carcinomas (WDTCs), and 10 high-grade thymic carcinomas (HGTCs), according to the Muller-Hermelink histologic classification. The HGTC showed the highest apoptotic index of the tumor cells. The apoptotic indices of the thymomas and WDTCs were significantly lower than those of the HGTCs (P < .001), but there was no significant difference among each type of thymoma and the WDTCs. bcl-2 protein was expressed in the tumor cells of the medullary-type thymomas and of the HGTCs but not in the other types of thymoma or WDTC. Fas antigen was negative in the HGTCs but was predominantly expressed in thymomas and WDTCs with advanced clinical stages (P < .05). From the immunostaining pattern, WDTC might be more likely to belong to the class of thymoma than to the category of HGTC. PMID- 9346178 TI - A prognostic model of survival in surgically resected squamous cell carcinoma of the lung using clinical, pathologic, and biologic markers. AB - The biologic behavior of tumoral cells plays a significant role in the progression of the neoplasia, because 30 to 35% of patients with Stage I squamous cell carcinoma relapse. The present study was designed to determine whether age, pathologic parameters, DNA ploidy, and a cell proliferation index (the area of nucleolar organizer regions, AgNOR), could be used to predict survival in patients who undergo resection for limited squamous cell carcinoma of the lung. For histopathologic analysis, the parameters of histologic grading, pleural involvement, vascular invasion, and residual disease were considered. The cell proliferation index was evaluated by mitotic index, AgNOR quantification, and DNA ploidy by means of digital image analysis. Fifty-two patients (median age, 60 yr +/- 8.6 yr) were staged according to the TNM staging system. Cox univariate analysis showed that stage, residual disease, vascular invasion, histologic grading, DNA ploidy, and AgNOR were significant predictors of survival. Many of the univariate predictors of cancer death, however were eliminated when Cox multivariate models were computed. The variable that exhibited the most robust predictive value for overall survival was AgNOR. We conclude that measurement of cell proliferation might serve as a prognostic marker in squamous cell carcinoma of the lung. PMID- 9346179 TI - Clear cell tumor of the lung: an immunohistochemical and ultrastructural study supporting a pericytic differentiation. AB - Clear cell tumor ("sugar tumor") of the lung is a rare benign lesion with unclear histogenesis. It is composed of large cells with a clear cytoplasm rich in glycogen, blended with an abundant network of sinusoid-type vessels. We report two cases of sugar tumor, one of these lacking clearly demonstrable glycogen storage. In both, the tumor cells lacked keratin expression and were positive for vimentin and HMB 45, an antibody recognizing perivascular or myoid cell proliferation such as lymphangioleiomyomatosis and angiomyolipoma. The tumor cells were also immunoreactive for an endothelial cell marker, CD 34, but negative for Factor VIII or smooth muscle actin. Intercellular deposition of basal-like material was immunostained with Type IV collagen. At ultrastructural examination of one of these cases, tumor cells showing features of pericytes or poorly differentiated perivascular leiomyocytes encased in basement material were observed in close association with endothelial cells; their cytoplasm contained numerous membrane-bound glycogen and pinocytic vesicles. We conclude that on the basis of immunohistochemical and ultrastructural phenotype, sugar tumor presents pericytic features and that glycogen storage is not a constant feature of these benign tumors. PMID- 9346180 TI - The effect of decalcification on in situ hybridization. AB - In situ hybridization (ISH) for mRNA polyadenylated sequences was performed on 25 non-neoplastic and neoplastic decalcified, paraffin-embedded tissues using a poly d(T) oligonucleotide probe to assess the efficacy of this molecular diagnostic tool on decalcified tissue samples. Three commercially available decalcifying agents were used, including one EDTA-based solution (Versenate) and two hydrochloric acid-based solutions (S/P Decal, RBD). Before decalcification, the tissues were fixed in formalin for 6, 24, and 72 hours, respectively. The results of ISH performed on decalcified tissues were compared with the results from the nondecalcified control samples for each tissue using a numeric scoring system (0, negative; 4, strong positivity equal to control; 5, stronger than control). There was generally excellent reactivity when using Versenate (mean, 4.15), good reactivity with S/P Decal (mean, 3.17), and fair-to-poor reactivity with RBD (mean, 1.69) (all P values < .0001). The length of time in formalin did not affect the outcome of ISH on these tissues. We conclude that ISH can be performed with success on decalcified, paraffin-embedded tissues when using Versenate, an EDTA-based agent. Although accurate results might be obtained with S/P Decal and RBD, caution should be exercised when using these two hydrochloric acid-based solutions because they might produce false-negative results. PMID- 9346181 TI - Intrasinusoidal bone marrow involvement by splenic lymphoma with villous lymphocytes: a helpful immunohistologic feature. AB - Splenic lymphoma with villous lymphocytes (SLVL) is a chronic monoclonal B-cell lymphoproliferative disorder characterized by massive splenomegaly and typical villous lymphocytes in the peripheral blood (PB). The diagnosis of SLVL relies on blood smear examination, phenotypic features, and marginal zone involvement of the spleen. The histologic pattern of bone marrow (BM) involvement has not been well characterized. We report four cases associated with a peculiar intrasinusoidal BM involvement. This intrasinusoidal pattern was highlighted by immunostaining that also showed the cytoplasmic projections of villous lymphocytes within routinely fixed and decalcified BM biopsy specimens. Therefore, a simple immunohistochemical analysis of BM involvement would help to identify SLVL. Combined with cytologic and immunophenotypic evaluation of marrow and blood smears, this intravascular pattern might be helpful in differentiating SLVL from hairy cell leukemia and its variant. Whether this peculiar intravascular pattern combined with cytologic evaluation represents a practical alternative to the diagnostic splenectomy must be confirmed by extensive studies focusing on this immunohistochemical criterion. PMID- 9346182 TI - bcl-2 protein expression in gastric remnant mucosa and gastric cancer 15 or more years after partial gastrectomy. AB - Partial gastrectomy is a risk factor for subsequent gastric cancer. The genetic alterations associated with malignant transformation, however, are poorly understood. Ninety-eight biopsies from 22 patients with benign gastric mucosa (BGM) at least 15 years after gastrectomy and resected specimens from 13 patients with postgastrectomy stump cancer (GC), were evaluated for immunohistochemical expression of bcl-2 oncogenic protein and correlated with the presence of dysplasia and subtypes of intestinal metaplasia (IM), categorized using high-iron diamine-alcian blue and alcian blue-periodic acid-Schiff stains. In BGM patients, 91% had chronic gastritis with atrophy, 18% showed complete (Type I) IM, 36% showed incomplete (Type II) IM, and 45% Type III IM. Twelve biopsy specimens from nine BGM patients showed mild-to-moderate dysplasia. Increased bcl-2 expression was present in 27% of BGM patients, with a significant association with increasing grade of IM: 20% in specimens with Type I IM, 30% with Type II, and 40% with Type III (P = .01). bcl-2 overexpression was more often present in the area of the anastomosis than in the body or fundus (P = .06). Of GC patients, 15% had Type II IM and 85% Type III IM. Moderate-to-severe dysplasia was present in adjacent benign mucosa in 46%. bcl-2 was present in 54% of GCs, and increased expression was detected in the adjacent benign mucosa in 60%. bcl-2 expression did not correlate with the presence or degree of dysplasia in either BGM or GC patients. bcl-2 protein is frequently expressed in GC. Increased expression is observed in mucosa adjacent to tumor and, to a lesser extent, in biopsy specimens of BGM, often associated with Type III IM. These findings suggest a possible role for the bcl-2 proto-oncogene in malignant progression. PMID- 9346183 TI - Solitary fibrous tumor of soft tissue: a report of 15 cases, including 5 malignant examples with light microscopic, immunohistochemical, and ultrastructural data. AB - We describe 15 soft tissue solitary fibrous tumors (SFTs) occurring in patients 24 to 78 years old (average, 50.6 yr). Ten tumors were benign and arose in the head and neck area (three tumors), thigh (two), vulva (two), upper arm (one), lower leg (one), and retroperitoneum (one). Five tumors were histologically malignant and arose in the thigh (two), abdominal wall (one), buttock (one), and retroperitoneum (one). All of the tumors were grossly well circumscribed. The benign tumors measured from 2 to 10 cm (average, 4.8 cm) and the malignant ones from 3 to 5.5 cm (average, 4.3 cm) in greatest diameter. Microscopically, the benign tumors showed areas of hypercellularity with variable amounts of collagenous and myxoid stroma; one had amianthoid fibers. The malignant tumors were composed of cytologically atypical cells enmeshed in a collagenous or myxoid extracellular matrix. Ultrastructural study of three benign and three malignant tumors showed fibroblastic differentiation; one benign tumor showed myofibroblastic differentiation. Immunohistochemically, all of the tumors examined were immunoreactive for vimentin, and seven of nine were positive for CD34, including all of the malignant ones. There was focal staining for muscle actin in two benign tumors and for Leu-7 in one benign tumor; there was no staining for cytokeratin, desmin, S-100 protein, epithelial membrane antigen, or smooth muscle actin in any of the examined tissues. Follow-up was available for eight patients for 6 to 21 months (average, 12 mo). No tumor recurred locally or metastasized. The SFTs reported herein support the experiences of others who recently described these tumors in the somatic soft tissues. In addition, our series highlights the occurrence of malignant SFTs in the soft tissues. SFTs should be separated from other spindle cell sarcomas, with which they can be confused. PMID- 9346185 TI - Techniques in human airway inflammation: differences in plastic-embedded and snap frozen sections for CD3, CD4, and CD8 immunostaining of bronchial biopsy specimens. AB - Today, the quantification of inflammatory cells in human airway biopsies might be facilitated by better morphologic resolution provided by special resin (plastic) embedding techniques. The present study compares the numbers of CD3-, CD4-, and CD8-positive cells in glycolmethacrylate-embedded versus snap-frozen biopsy specimens of normal bronchial mucosa in 10 patients with various pulmonary diseases. In general, larger numbers of CD3-, CD4-, and CD8-positive cells were counted in snap-frozen specimens than in plastic-embedded ones. Loss of antigenic properties during storage of plastic-embedded tissue (blocks) might have contributed to the weak correlation between both methods. An additional study showed that the number of CD3-, CD4-, and CD8-positive cells decreased significantly within a few months after embedding in glycolmethacrylate. Therefore, we recommend processing glycolmethacrylate-embedded specimens as soon as possible. For standard evaluation of established inflammatory cell parameters such as CD3, CD4, CD8, and EG2, frozen tissue is preferable because of the ease of the method and its reliable cell counting. Because glycolmethacrylate-embedded tissue shows superior morphologic resolution, under strict rules, this method seems attractive for the study, in particular, of cell-cell and cell-matrix relationships. PMID- 9346184 TI - Immunohistochemical diagnosis of typhus rickettsioses using an anti lipopolysaccharide monoclonal antibody. AB - A monoclonal antibody directed against an epitope on the lipopolysaccharide of typhus-group rickettsiae was developed for the purpose of detecting this heat stable, proteinase-resistant antigen in formalin-fixed, paraffin-embedded tissues. Rickettsia prowazekii organisms were identified in endothelium and macrophages in sections of the brains of three Egyptian men who died of epidemic louse-borne typhus in Cairo during World War II and in the brain from a recent case of typhus fever acquired in Burundi. R. typhi organisms were identified in endothelial cells from a fatal case of murine typhus and in experimentally infected mice. This approach is applicable not only to the study of archival tissues and experimental animal models but also could be used to establish a timely diagnosis of typhus-group rickettsiosis by immunohistochemical examination of cutaneous biopsies of rash lesions during the acute stage of illness. PMID- 9346186 TI - Primary osteorhabdomyosarcoma (malignant mesenchymoma) of bone: a case report and review of the literature. AB - Primary malignant mesenchymoma of bone is a rare neoplasm consisting of two or more unrelated malignant mesenchymal components other than fibrosarcoma or malignant fibrous histiocytoma. The literature reports fewer than 15 cases, most of which were composed of osteosarcoma and liposarcoma. We report an exceedingly rare case of primary malignant mesenchymoma of bone composed of osteosarcoma and rhabdomyosarcoma (osteorhabdomyosarcoma), arising in the right proximal tibia of a 21-year-old woman. We review the literature and compare primary malignant mesenchymoma of bone with dedifferentiated chondrosarcoma and conventional intramedullary osteosarcoma. PMID- 9346187 TI - Hepatoid carcinoma of the lung with production of alpha-fetoprotein and abnormal prothrombin: an autopsy case report. AB - Alpha-fetoprotein (AFP) is a useful tumor marker for the diagnosis of hepatic and testicular tumors. Several cases of AFP-producing lung cancer have been reported. We present here a patient with AFP-producing primary lung carcinoma, which showed high values of serum AFP (100,000 ng/mL). The concanavalin A nonbinding fraction rate of AFP was 15%. Gross and microscopic features of the lung carcinoma bore a striking resemblance to those of hepatocellular carcinoma. According to the histologic classification of lung tumor, this case was large cell carcinoma with prominent hepatoid differentiation. Immunohistochemically, we detected AFP in the cytoplasm of the neoplastic cells. We also detected another useful tumor marker for the diagnosis of hepatocellular carcinoma, i.e., des-gamma-carboxy prothrombin (protein induced by vitamin K absence or the absence of antagonist-II [PIVKA-II]), in serum using an enzyme immunoassay and in tumor cells by immunohistochemical analysis. PMID- 9346216 TI - Current status of chemotherapy in gynecologic cancer. PMID- 9346188 TI - Nodular pulmonary immunoglobulin light chain deposits with coexistent amyloid and nonamyloid features in an HIV-infected patient. AB - Isolated nodular pulmonary amyloidosis is a rare condition characterized by localized deposits of immunoglobulin (Ig) light chain amyloid. Nonamyloid nodular light chain deposits in lungs can occur in systemic light chain deposition disease. Both amyloid and nonamyloid light chain deposits have been described at separate sites in the same or different organs but rarely in lungs. We report the clinical, radiologic, and pathologic findings in a drug user infected with the human immunodeficiency virus who had multinodular pulmonary Ig light chain deposits consisting of both amyloid and nonamyloid granular morphologic features. The deposits, closely associated with numerous plasma cells, had a unique histochemical and ultrastructural profile, with intermixed Congo red-positive fibrillar amyloid and Congo red-negative granular nonamyloid components. Immunohistochemical and immunoelectron microscopic studies showed reactivity of both the fibrillar and granular deposits for kappa and lambda light chains but not heavy chains. There was no evidence of restricted clonality of local or bone marrow plasma cells, serum or urine monoclonal protein, or secondary causes of amyloidosis. The amyloid deposits (but not the nonamyloid deposits) were reactive with antibody to amyloid rho component. There was no staining for other types of amyloid, i.e., amyloid A or transthyretin. The relationship between pulmonary amyloidosis, infection with the human immunodeficiency virus, and illicit drug use is unknown. We conclude that the nodular pulmonary light chain deposits with both amyloid and nonamyloid morphologic features are related to local plasma cell proliferation and that the fibrillar and nonfibrillar components most likely result from different conformations of the Ig light chains. PMID- 9346217 TI - ICON 2 and ICON 3 data in previously untreated ovarian cancer: results to date. AB - The second International Collaborative Ovarian Neoplasm study (ICON 2), was a large, international randomized study of cyclophosphamide/doxorubicin/cisplatin (CAP) versus single-agent carboplatin in patients with previously untreated ovarian cancer. Patients in the CAP arm received 500 mg/m2 cyclophosphamide, 50 mg/m2 doxorubicin, and 50 mg/m2 cisplatin. Carboplatin was dosed to an area under the concentration-time curve of 5. Chemotherapy was given every 3 weeks for a total of 6 months for each regimen. Results of a 1995 interim analysis in 1,377 patients showed that overall, the total dose received in the CAP group was greater than 75% of the planned dose, with 75% of patients receiving greater than 87%, greater than 80%, and greater than 83% of the planned doses of cisplatin, doxorubicin, and cyclophosphamide, respectively. Of carboplatin patients, 75% received greater than 1,450 mg/m2 (of a median planned dose of 1,800 mg/m2). There were significant differences in grades 3/4 toxicity between the two arms: leukopenia occurred in 34% of CAP patients versus 10% of carboplatin patients, alopecia in 70% versus 3%, nausea and vomiting in 21% versus 9%, and mucositis in 21% versus 0%, respectively. However, thrombocytopenia was more frequent in the carboplatin group (16% v 7%). At the 1996 analysis, 1,526 patients had been entered, 1,498 had been randomized, and 740 had progressed or died. The interim conclusions were that although the more toxic CAP regimen may improve progression free survival by a small amount, there was no evidence of any survival benefits or difference within the subgroups. Given that a sufficient number of patients had been accrued to ICON 2 and that starting accrual for ICON 3 markedly slowed accrual to ICON 2, the trial was closed. The full analysis of ICON 2 should be available in the summer of 1997. ICON 3 was designed to consider the role of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) in previously untreated ovarian cancer. Patients received paclitaxel at a dose of 175 mg/m2 (3 hour infusion) in combination with carboplatin dosed to an area under the concentration-time curve of 5 (based on chromium ethylenediamine tetraacetic acid) or 6 (based on calculated creatinine clearance). The control arm was either CAP or carboplatin. Patients were randomized 2:1 in favor of the control arm. In all, 1,070 patients have been entered to date. At the first planned interim analysis, in April 1996 in 434 patients, it was too early to provide efficacy data. The plan was to continue accruing to the trial to a maximum of 2,000 patients, with review in mid-1997, when the events for analysis will be virtually doubled and the data can be interpreted in light of the results of the Intergroup study (European Organization for Research and Treatment of Cancer, the National Cancer Institute of Canada, and the Scottish study) and Gynecologic Oncology Group trial 132. PMID- 9346218 TI - Carboplatin plus paclitaxel in the treatment of gynecologic malignancies: the Cleveland Clinic experience. AB - To examine the toxicity profile and antineoplastic activity of carboplatin (area under the concentration-time curve of 4 to 7.5) plus 3-hour infusional paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) (135 or 175 mg/m2) in women with advanced gynecologic malignancies, we retrospectively reviewed the experience of the Gynecologic Cancer Program at The Cleveland Clinic with this combination chemotherapy regimen. To date, 92 patients (median age, 67 years) have received a total of 460 courses (median number per patient, six) of this two drug combination. The initial paclitaxel dose was 175 mg/m2 and the carboplatin area under the concentration-time curve was > or = 5 in 72% and 73% of patients, respectively. The major toxicity was neutropenia (grade 4 in 9% of patients), resulting in two febrile episodes and a single septic death. Grade 4 thrombocytopenia and grade 3 peripheral neuropathy were noted in one and two patients, respectively. Twelve patients (13%) experienced at least one episode of paclitaxel-associated hypersensitivity, but all were able to continue with the treatment program. Of the 62 patients with ovarian cancer or primary peritoneal carcinoma with carbohydrate antigen-125 levels > or = 60 U/mL before the initiation of chemotherapy, 74% exhibited a > or = 90% decline in the tumor marker following treatment. We conclude that the combination of carboplatin and 3 hour infusional paclitaxel can be administered in the outpatient setting with a highly acceptable toxicity profile and with major activity in patients with ovarian cancer and primary carcinoma of the peritoneum. PMID- 9346219 TI - Efficacy and safety of the combination paclitaxel/carboplatin in patients with previously treated advanced ovarian carcinoma: a multicenter French Groupe des Investigateurs Nationaux pour l'Etude des Cancers Ovariens phase II study. AB - The French Groupe des Investigateurs Nationaux pour l'Etude des Cancers Ovariens (GINECO) conducted a multicenter phase II study of carboplatin and paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) to evaluate the efficacy and side effects of this combination in pretreated advanced ovarian cancer. Patients with progressive ovarian carcinoma during or after platinum-based chemotherapy received paclitaxel 175 mg/m2 intravenously over 3 hours followed by intravenous carboplatin over 30 minutes every 4 weeks. The dose of carboplatin was calculated using a projected area under the concentration-time curve of 5 mg/mL x min. Of the 50 patients entered, 50 were evaluable for toxicity and 42 for response. There were eight complete and 10 partial responses, for an overall response rate of 43% (95% confidence interval, 28% to 56%). Overall response rates in platinum refractory patients and in those with early (> or = 3 and < 12 months) and late (> or = 12 months) relapse was 28%, 33%, and 71%, respectively. Median response duration, progression-free survival, and overall survivals were 8, 6, and 14 months, respectively. The most frequent and severe toxicity was myelosuppression. Grades 3 and 4 neutropenia occurred in 30% and 23% of cycles, and granulocyte colony-stimulating factor was administered in 6%. Only one case of neutropenic fever was observed. Grades 3 and 4 thrombocytopenia occurred in 3% and 1% of cycles, respectively. Alopecia and moderate nausea or vomiting were frequent. Transitory peripheral neuropathy was present in 45% of patients but was severe in only one patient. One early death was observed due to progressive disease and possibly to therapy. The combination of paclitaxel 175 mg/m2 as a 3-hour infusion and carboplatin dosed to an area under the concentration-time curve of 5 is an effective therapy in patients previously treated with platinum-based chemotherapy and may be administered safely to outpatients who relapse after one or two lines of chemotherapy. PMID- 9346220 TI - Paclitaxel (175 mg/m2 over 3 hours) with cisplatin or carboplatin in previously untreated ovarian cancer: an interim analysis. AB - The side effects of cisplatin (75 mg/m2) in combination with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) (175 mg/m2 over 3 hours) are expected to be more severe and frequent than those of carboplatin (area under the concentration-time curve of 5) in combination with the same dose of paclitaxel, but the combinations are expected to be equally effective. A disadvantage of the cisplatin-based regimen is that patients need to be admitted to the hospital. The carboplatin regimen can be administered to outpatients. We tested both combinations administered every 3 weeks in a randomized phase III study in patients with previously untreated epithelial ovarian cancer. An interim analysis for toxicity was performed in 145 patients shortly after study closure. We observed a difference in the incidence of nausea, vomiting, and neurotoxicity favoring the women treated with the carboplatin regimen, but this regimen caused more myelotoxicity. Maturation of the study is awaited before survival data can be analyzed and final conclusions can be drawn. PMID- 9346221 TI - Dose-escalated paclitaxel in 1-hour infusion with a fixed dose of cisplatin in previously untreated advanced ovarian cancer: a phase II trial of the Spanish Group for Ovarian Cancer. AB - This phase II trial was planned to study the efficacy and toxicity of a fixed dose of cisplatin plus paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) given over 1 hour with intrapatient dose escalation. Patients with advanced epithelial ovarian cancer (stages IIB-IV); Eastern Cooperative Oncology Group performance status < or = 2; normal renal, liver, and bone marrow function; and evaluable residual disease after debulking surgery were accrued. Paclitaxel was given over 1-hour infusion and dose was escalated from 175 to 200 and 225 mg/m2 if nadir neutrophil counts were > or = 1000/microL, platelets were > or = 100,000/microL, and neurotoxicity was less than grade 2. Cisplatin was given after paclitaxel at a fixed dose of 80 mg/m2. Six courses at 3-week intervals were planned. From May 1995 to August 1996, 68 patients were entered. Paclitaxel could not be escalated in six patients, another six received up to 200 mg/m2, and 45 received 225 mg/m2. Three hundred seventy-five courses were given: 27.7% at 175 mg/m2, 19.2% at 200 mg/m2, and 53.1% at 225 mg/m2. All patients were evaluable for toxicity, and 67 were evaluable for response. Thirty-five patients had a complete clinical response (51.4%), 20 had a partial response (29.4%), six had stable disease (8.9%), and six progressed on therapy (8.9%). Overall response rate was 80.8 (95% confidence interval, 71.3% to 90.1%). Second-look laparotomy was performed in 32 patients, and 20 of them (62.5%) had a pathologic complete remission. Grade 3 or 4 neutropenia was seen in 26 patients (38%), but only one had fever. Severe thrombocytopenia was not seen. Peripheral neurotoxicity (grade 1, 39.7%; grade 2, 42.6%; and grade 3, 8.8%) was dose-limiting. It is too early to report on time to progression and survival, and these data are not yet available. This combination of cisplatin with escalating doses of paclitaxel is feasible and very active, but the high incidence of peripheral neurotoxicity may limit its use. PMID- 9346222 TI - Carboplatin/paclitaxel versus cisplatin/paclitaxel as first-line chemotherapy in advanced ovarian cancer: an interim analysis of a randomized phase III trial of the Arbeitsgemeinschaft Gynakologische Onkologie Ovarian Cancer Study Group. AB - Since publication of the results of the Gynecologic Oncology Group III study, the combination of cisplatin/paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) has been widely adopted as standard treatment for advanced ovarian cancer. Further attempts to optimize first-line chemotherapy with platinum and taxanes include the substitution of cisplatin with carboplatin, individualization of the carboplatin dose by calculating it according to the area under the concentration-time curve, and reduction of paclitaxel infusion duration. These attempts have led to the initiation of several phase I/II trials evaluating the combination of carboplatin/paclitaxel. The promising results of these small studies have prompted the initiation of three phase III trials comparing carboplatin/paclitaxel with the standard combination of cisplatin/paclitaxel. The interim analysis after 15 months' accrual of the prospectively randomized German Arbeitsgemeinschaft Gynakologische Onkologie (AGO) study is presented here. As of January 1997, 518 of 660 planned patients have been recruited. The interim analysis is based on data from 449 evaluable patients. The preliminary data indicate that hematologic toxicity occurred more frequently in arm A (carboplatin/paclitaxel), whereas nonhematologic toxicity occurred slightly more frequently in arm B (cisplatin/paclitaxel). Dose-intensity analysis did not reveal cumulative dose reductions or an increased need for colony-stimulating factors over subsequent courses in both arms. Forty-four patients with measurable disease following surgery completed chemotherapy and were evaluable for response, which remains blinded at this time and is reported for the group as a whole. So far, there have been 18 complete responses (41%) and 15 partial responses (34%), for an overall response rate of 75%. Retrospective comparison reveals no significant difference in response rates between patients in the cisplatin/paclitaxel arm of Gynecologic Oncology Group III and those in the Arbeitsgemeinschaft Gynakologische Onkologie study. Overall, this interim analysis did not reveal any reason for an early termination of this study. Accrual is ongoing and is expected to be completed this year. PMID- 9346223 TI - Paclitaxel in platinum-resistant ovarian cancer patients. Argentine Multicenter Taxol Group. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company; Princeton, NJ) is an antineoplastic agent that inhibits microtubular function and has shown efficacy in several solid tumors, mainly ovarian tumors, in which 20% to 40% response rates in previously treated patients were observed. We conducted a study to assess survival, response rate, and toxicity associated with paclitaxel treatment in patients with advanced ovarian cancer resistant to platinum therapy. Between September 1994 and November 1996, 38 patients were admitted for study and 37 were evaluable. All had disease progression or relapse within 1 year of receiving platinum-containing first-line chemotherapy. Mean age was 59 years (range, 30 to 75 years), all had bulky disease, and 18 showed increased carbohydrate antigen 125 at admission. They were treated every 3 weeks with paclitaxel 175 mg/m2 as a 3-hour infusion, preceded by standard premedication. Response rate was 51.3%, with a median response duration of 10.0 months and a median survival rate of 16.8 months. Mild to moderate hematologic toxicity was observed with only one episode of grade 4 neutropenia, without fever. Gastrointestinal toxicity was moderate and peripheral neuropathy was mild, except for two patients who had concomitant pathologies or previous treatment, which might have caused some neuropathy. We concluded that paclitaxel given as a 3-hour infusion was easily administered for ambulatory treatment, with mild to moderate toxicity and promising results based on rate and duration of response as well as survival. PMID- 9346224 TI - Paclitaxel with carboplatin versus paclitaxel with carboplatin alternating with cisplatin as first-line chemotherapy in advanced epithelial ovarian cancer: preliminary results of a Hellenic Cooperative Oncology Group study. AB - Ninety previously untreated patients with advanced epithelial ovarian cancer (International Federation of Gynecology and Obstetrics stages IIC, III, and IV) were randomized, after initial cytoreductive surgery, to receive paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) 175 mg/m2 as a 3-hour infusion with either carboplatin at an area under the concentration-time curve of 7 (group A) or carboplatin at an area under the concentration-time curve of 7 on courses 1, 3, and 5, alternating with cisplatin 75 mg/m2 on courses 2, 4, and 6 (group B). Treatment was given every 3 weeks, up to a total of six courses. Sixty one patients (33 and 28 patients in groups A and B, respectively) had residual disease after the initial cytoreductive surgery. Patients with measurable or evaluable disease had a high overall response (82% v 57%). A 52% and 39% complete response rate for groups A and B, respectively, with no statistically significant difference between the groups was also observed. With a median follow-up of 12 months (range, 0.33 to 24 months), 29 patients have progressed (18 and 11 in groups A and B, respectively), and 13 have died (seven and six, respectively). Median time to progression was 20.36 months (range, 0.20 to 23.54 months) for group A, whereas this has not yet been reached for group B. Median survival has not yet been reached, but there is no significant difference between the two groups (P = .6972). Treatment was generally well tolerated. Grade 3 and 4 neutropenia was 20% and 32% for groups A and B, respectively, while grade 3 and 4 thrombocytopenia was 4% and 7%, respectively, with no significant difference between the two groups. In conclusion, both combinations seem very active for the treatment of advanced epithelial ovarian cancer and are associated with acceptable toxicity. PMID- 9346225 TI - Salvage weekly paclitaxel in recurrent ovarian cancer. AB - The objectives of this study were to determine the toxicity and phase II dose of weekly paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) administration and to describe our initial experience with salvage weekly intravenous paclitaxel in women with advanced recurrent ovarian carcinoma. We conducted a phase I trial of paclitaxel administered as a 1-hour infusion in advanced ovarian cancer patients, using doses of 40 to 100 mg/m2/wk. As a follow up study, we retrospectively reviewed the medical records of 45 patients with advanced, recurrent epithelial ovarian cancer treated between January 1, 1996, and October 30, 1996, with single-agent weekly intravenous paclitaxel (60 to 100 mg/m2, 1-hour infusions). Response for patients with measurable disease was based on improvements in physical examination or a greater than 50% reduction in the perpendicular diameters of all lesions in serial computed tomography. Since many of the patients did not have measurable disease, some evaluations of response to therapy were based on decline in serum carbohydrate antigen-125 according to published criteria. In our phase I study, 18 patients received 194 weekly courses of therapy at doses of 40, 50, 60, 80, and 100 mg/m2. The dose-limiting toxicity was defined as two or more patients with treatment delay or grade 3 toxicity (National Cancer Institute common toxicity criteria) at the same dose level. Treatment was delayed in two of three patients (for neutrophil counts < 1,500/microL at the 100 mg/m2 dose level); therefore, a phase II dose of 80 mg/m2/wk is recommended. No patient required hospitalization for neutropenia/fever. In the retrospective cohort review, the median patient age was 55 years (range, 32 to 79 years). All patients were heavily pretreated with multiple systemic chemotherapy regimens, including paclitaxel, with a median of four regimens (range, one to eight) before receiving weekly paclitaxel. Patients received a median of nine cycles of weekly paclitaxel (range, three to 37), with a median interval of 8 months (range, 1 to 32 months) between the last paclitaxel treatment and the institution of weekly therapy. Response was noted in 13 of 45 (28.9%) patients, with a median of seven treatments to achieve response. Chemotherapy was generally well tolerated, with treatments completed on a weekly schedule and only one hospitalization for nadir fever. We conclude that weekly intravenous paclitaxel is an active and well-tolerated regimen in heavily pretreated women with recurrent ovarian carcinoma. Prior therapy with paclitaxel does not preclude response to this regimen. A phase II trial of weekly paclitaxel in paclitaxel-refractory patients is under way. PMID- 9346227 TI - Paclitaxel, estramustine, and etoposide in the treatment of hormone-refractory prostate cancer. AB - We previously developed a novel and effective therapy for hormone-refractory prostate cancer using the agents estramustine and etoposide. Although neither of these agents alone is effective in the treatment of advanced, hormone-refractory prostate cancer, we predicted their activity when used in combination based on preclinical assays, and then demonstrated their effectiveness in a phase I-II clinical trial, where they were shown to produce a 50% complete and partial response rate in patients with bidimensionally measurable disease. In preclinical studies, we had demonstrated that estramustine and etoposide interact with the nuclear matrix, which is the site of DNA replication. Expanding these investigations, we determined that paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), a microtubule inhibitor, interacts with estramustine and etoposide, and the combination of these three agents had significant preclinical activity against androgen-independent prostate cancer cells. These studies led us to conduct a phase II clinical trial of paclitaxel, estramustine, and etoposide in patients with hormone-refractory prostate cancer. Preliminary results demonstrate that this is an active regimen, with 57% of patients demonstrating a response to therapy as measured by a decrease in pretreatment prostate-specific antigen levels of greater than 50%. PMID- 9346226 TI - Phase I trial of paclitaxel, carboplatin, and methotrexate with granulocyte colony-stimulating factor and leucovorin in advanced transitional cell carcinoma. AB - Advanced transitional cell carcinoma (TCC) of the urothelial tract is usually fatal despite high response rates to platinum-based chemotherapy regimens. Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) has demonstrated marked single-agent activity in TCC, and combinations of carboplatin and paclitaxel have been well tolerated in other solid tumors. Methotrexate is also active in TCC. Due to unexpectedly severe myelosuppression and mucositis when methotrexate and paclitaxel were combined, we undertook a phase I trial of paclitaxel, carboplatin, and escalating doses of methotrexate with granulocyte colony-stimulating factor and leucovorin support in advanced TCC to determine the feasibility of this combination. Nineteen previously untreated patients with locally advanced or metastatic TCC were eligible. Median age was 62 years. In sequence, paclitaxel 200 mg/m2 (3-hour infusion), carboplatin dosed to an area under the concentration-time curve of 6 mg/mL x min, and methotrexate 10 mg/m2, increasing in 10-mg/m2 increments, were administered on day 1 every 21 days. Granulocyte colony-stimulating factor 300 microg/d or 480 microg/d (in patients <60 kg or >60 kg, respectively) was administered on days 2 through 11 and leucovorin 15 mg orally every 6 hours for 3 days. At this time, the methotrexate dose has been escalated to 50 mg/m2. There were no dose-limiting toxicities in cycle 1. Sixty-eight cycles have been administered (range, one to eight cycles; median, three cycles). Significant hematologic toxicity including neutropenic sepsis (two episodes) occurred in subsequent cycles, but was infrequent. The major nonhematologic toxicity was neuropathy. Sixteen patients are evaluable for response. One patient has achieved a complete response, seven are partial responders, seven have stable disease, and one progressed on therapy. The overall response rate is 50% (95% confidence interval, 25% to 75%). The combination of paclitaxel, carboplatin, and methotrexate holds promise to be well tolerated and active in advanced TCC. PMID- 9346228 TI - A trial of outpatient paclitaxel and carboplatin for advanced, recurrent, and histologic high-risk endometrial carcinoma: preliminary report. AB - The purpose of this study was to evaluate the combination of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and carboplatin in patients with endometrial cancer known to be resistant to standard therapy. Subjects were taken from three groups: (1) recurrent or persistent disease following surgery and/or radiation, (2) advanced disease at diagnosis, and (3) high-risk histology. The combination of carboplatin (pharmacologically dosed at an area under the concentration-time curve of 5) and paclitaxel (135 to 175 mg/m2 over 3 hours) was given intravenously every 4 weeks for eight courses. Data about response, overall and progression-free survival, and toxicity were collected. Response and toxicity were evaluated by physical examinations, x-ray films, and blood tests. Twenty patients have participated to date, including eight considered evaluable for response. Due to limited follow-up, survival and progression-free intervals are not yet assessable. Of patients with measurable disease, five of eight (63%) have had significant reduction in the size of evaluable tumor masses, constituting a partial response. Although two patients had clinical and radiographic complete responses, occult disease was found at surgery. There were no complete responders. Fifteen patients had grade 3 or 4 hematologic toxicity, but none had neutropenic fever or hospitalization for sepsis. One patient was taken off study for grade 3 neuropathy. There was one possible treatment-related death. In this preliminary report, this combination is active against tumors of the endometrium, with acceptable levels of toxicity. Further follow-up will be required to determine the duration of response and whether progression-free and overall survival are influenced by treatment with these drugs. PMID- 9346229 TI - Paclitaxel in salvage therapy for germ cell tumors. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) as a single agent showed antitumor activity in patients with germ cell tumors resistant to combination cisplatin-containing chemotherapy and has been included in two risk directed first-line combination salvage programs. Patients with relapsed germ cell tumors originating from a testis primary site were treated in a phase I/II trial of conventional-dose paclitaxel (given at 175, 215, and 250 mg/m2 dose levels), ifosfamide, plus cisplatin. Ten (63%) of 16 assessable patients achieved a complete response, eight (50%) of which remain durable at a median follow-up of 16 months. The 250 mg/m2 dose of paclitaxel was chosen for the phase II trial and accrual continues. In a second trial, patients with cisplatin-resistant germ cell tumors and unfavorable prognostic features (previous incomplete response to first line chemotherapy or an extragonadal primary site) were treated with a dose intensive program consisting of rapid recycling of paclitaxel plus ifosfamide followed by carboplatin plus etoposide with stem cell support. The target carboplatin dose was based on the Calvert formula and escalated from an area under the concentration-time curve of 12 to 32 mg/mL min among patient cohorts. Ten (56%) of 18 assessable patients achieved a complete response, and seven (39%) remain in complete response at a median follow-up of 11 months. Hematologic toxicity was moderately severe, but no treatment-related deaths have occurred. Accrual to the trial continues, and pharmacology studies for carboplatin are being correlated with the target area under the concentration-time curve by the Calvert formula. PMID- 9346230 TI - Future directions in the chemotherapy of ovarian cancer. AB - Clinical research in the treatment of epithelial ovarian cancer has entered a new phase. A pivotal Gynecologic Oncology Group study demonstrated that paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) plus cisplatin was superior to the previous standard therapy of cyclophosphamide plus cisplatin. Confirmatory trials in Europe have either been completed or are nearing an interim analysis. However, many questions still remain regarding what constitutes optimum chemotherapy in ovarian cancer. Clinical trials are addressing issues regarding the optimum use of paclitaxel, including dose, schedule, and duration of therapy. Several clinical trials throughout the world are comparing cisplatin plus paclitaxel versus a combination of carboplatin plus paclitaxel. In addition, the role of high-dose chemotherapy in ovarian cancer is being studied in important clinical trials in previously untreated patients or in patients who have had a response to initial therapy. These trials of high-dose chemotherapy with hematologic support will define the role, if any, of dose intensity either as part of initial therapy or as part of consolidation. A recent study has also demonstrated that chemotherapy with intraperitoneal cisplatin was superior to intravenous cisplatin in patients with optimal stage III ovarian cancer. However, additional confirmatory trials are needed to define the role of intraperitoneal therapy when combined with paclitaxel. A series of new agents have been identified with substantial activity in recurrent ovarian cancer, including topotecan, oral etoposide, gemcitabine, vinorelbine, and encapsulated doxorubicin. Future clinical trials will be addressing the incorporation of these agents into the up-front treatment of patients with advanced ovarian cancer. PMID- 9346231 TI - Eliminating all obstacles: regulated proteolysis in the eukaryotic cell cycle. PMID- 9346232 TI - Parsing the heart: genetic modules for organ assembly. PMID- 9346233 TI - KSHV and Kaposi's sarcoma: the end of the beginning? PMID- 9346234 TI - Reprogramming chemotaxis responses: sensory neurons define olfactory preferences in C. elegans. AB - Different olfactory cues elicit distinct behaviors such as attraction, avoidance, feeding, or mating. In the nematode C. elegans, these cues are sensed by a small number of olfactory neurons, each of which expresses several different odorant receptors. The type of behavioral response elicited by an odorant could be specified by the olfactory receptor or by the olfactory neuron in which the receptor is activated. The attractive odorant diacetyl is detected by the receptor protein ODR-10, which is normally expressed in the AWA olfactory neurons. The repulsive odorant 2-nonanone is detected by the AWB olfactory neurons. Transgenic animals that express ODR-10 in AWB rather than AWA avoid diacetyl, while maintaining qualitatively normal responses to other attractive and repulsive odorants. Animals that express ODR-10 simultaneously in AWA and AWB have a defective response to diacetyl, possibly because of conflicting olfactory inputs. Thus, an animal's preference for an odor is defined by the sensory neurons that express a given odorant receptor molecule. PMID- 9346235 TI - Nuclear translocation of extradenticle requires homothorax, which encodes an extradenticle-related homeodomain protein. AB - We show that homothorax (hth) is required for the Hox genes to pattern the body of the fruit fly, Drosophila melanogaster. hth is necessary for the nuclear localization of an essential HOX cofactor, Extradenticle (EXD), and encodes a homeodomain protein that shares extensive identity with the product of Meis1, a murine proto-oncogene. MEIS1 is able to rescue hth mutant phenotypes and can induce the cytoplasmic-to-nuclear translocation of EXD in cell culture and Drosophila embryos. Thus, Meis1 is a murine homolog of hth. MEIS1/HTH also specifically binds to EXD with high affinity in vitro. These data suggest a novel and evolutionarily conserved mechanism for regulating HOX activity in which a direct protein-protein interaction between EXD and HTH results in EXD's nuclear translocation. PMID- 9346236 TI - The homeobox gene GBX2, a target of the myb oncogene, mediates autocrine growth and monocyte differentiation. AB - The homeobox gene GBX2 was identified as a target gene of the v-Myb oncoprotein encoded by the avian myeloblastosis virus (AMV). GBX2 activation by c-Myb requires signal transduction emanating from the cell surface while the leukemogenic AMV v-Myb constitutively induces the GBX2 gene. Mutations in the DNA binding domain of AMV-Myb render it independent of signaling events and concomitantly abrogate the collaboration between Myb and CCAAT Enhancer Binding Proteins (C/EBP), which are involved in granulocyte differentiation. Ectopic expression of GBX2 in growth factor-dependent myeloblasts induces monocytic features and independence from exogenous cytokines, reflecting distinct features of AMV-transformed cells. Our results suggest that Myb or factors it interacts with contribute to hematopoietic lineage choice and differentiation in a signal transduction-dependent fashion. PMID- 9346237 TI - Multiple female reproductive failures in cyclooxygenase 2-deficient mice. AB - Cyclooxygenase (COX) is the rate-limiting enzyme in the synthesis of prostaglandins (PGs) and exists in two isoforms, COX-1 and COX-2. In spite of long-standing speculation, definitive roles of PGs in various events of early pregnancy remain elusive. We demonstrate herein that the targeted disruption of COX-2, but not COX-1, in mice produces multiple failures in female reproductive processes that include ovulation, fertilization, implantation, and decidualization. Using multiple approaches, we conclude that these defects are the direct result of target organ-specific COX-2 deficiency but are not the result of deficiency of pituitary gonadotropins or ovarian steroid hormones, or reduced responsiveness of the target organs to their respective hormones. PMID- 9346238 TI - F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex. AB - We have reconstituted the ubiquitination pathway for the Cdk inhibitor Sic1 using recombinant proteins. Skp1, Cdc53, and the F-box protein Cdc4 form a complex, SCFCdc4, which functions as a Sic1 ubiquitin-ligase (E3) in combination with the ubiquitin conjugating enzyme (E2) Cdc34 and E1. Cdc4 assembled with Skp1 functions as the receptor that selectively binds phosphorylated Sic1. Grr1, an F box protein involved in Cln destruction, forms complexes with Skp1 and Cdc53 and binds phosphorylated Cln1 and Cln2, but not Sic1. Because the constituents of the SCF complex are members of protein families, SCFCdc4 is likely to serve as the prototype for a large class of E3s formed by combinatorial interactions of related family members. SCF complexes couple protein kinase signaling pathways to the control of protein abundance. PMID- 9346239 TI - A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p. AB - In S. cerevisiae, the G1/S transition requires Cdc4p, Cdc34p, Cdc53p, Skp1p, and the Cln/Cdc28p cyclin-dependent kinase (Cdk). These proteins are thought to promote the proteolytic inactivation of the S-phase Cdk inhibitor Sic1p. We show here that Cdc4p, Cdc53p, and Skp1p assemble into a ubiquitin ligase complex named SCFCdc4p. When mixed together, SCFCdc4p subunits, E1 enzyme, the E2 enzyme Cdc34p, and ubiquitin are sufficient to reconstitute ubiquitination of Cdk phosphorylated Sic1p. Phosphorylated Sic1p substrate is specifically targeted for ubiquitination by binding to a Cdc4p/Skp1p subcomplex. Taken together, these data illuminate the molecular basis for the G1/S transition in budding yeast and suggest a general mechanism for phosphorylation-targeted ubiquitination in eukaryotes. PMID- 9346240 TI - Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. AB - Growth factors can promote cell survival by activating the phosphatidylinositide 3'-OH kinase and its downstream target, the serine-threonine kinase Akt. However, the mechanism by which Akt functions to promote survival is not understood. We show that growth factor activation of the PI3'K/Akt signaling pathway culminates in the phosphorylation of the BCL-2 family member BAD, thereby suppressing apoptosis and promoting cell survival. Akt phosphorylates BAD in vitro and in vivo, and blocks the BAD-induced death of primary neurons in a site-specific manner. These findings define a mechanism by which growth factors directly inactivate a critical component of the cell-intrinsic death machinery. PMID- 9346241 TI - The IkappaB kinase complex (IKK) contains two kinase subunits, IKKalpha and IKKbeta, necessary for IkappaB phosphorylation and NF-kappaB activation. AB - Recently we purified a 900 kDa cytokine-responsive IkappaB kinase complex (IKK) and molecularly cloned one of its subunits, IKKalpha, a serine kinase. We now describe the molecular cloning and characterization of IKKbeta, a second subunit of the IKK complex. IKKbeta is 50% identical to IKKalpha and like it contains a kinase domain, a leucine zipper, and a helix-loop-helix. Although IKKalpha and IKKbeta can undergo homotypic interaction, they also interact with each other and the functional IKK complex contains both subunits. The catalytic activities of both IKKalpha and IKKbeta make essential contributions to IkappaB phosphorylation and NF-kappaB activation. While the interactions between IKKalpha and IKKbeta may be mediated through their leucine zipper motifs, their helix-loop-helix motifs may be involved in interactions with essential regulatory subunits. PMID- 9346242 TI - GRASP65, a protein involved in the stacking of Golgi cisternae. AB - NEM prevents mitotic reassembly of Golgi cisternae into stacked structures. The major target of NEM is a 65 kDa protein conserved from yeast to mammals. Antibodies to this protein and a recombinant form of it block cisternal stacking in a cell-free system, justifying its designation as a Golgi ReAssembly Stacking Protein (GRASP65). One of the two minor targets of NEM is GM130, previously implicated in the docking of transport vesicles and mitotic fragmentation of the Golgi stack. GRASP65 is complexed with GM130 and is tightly bound to Golgi membranes, even under mitotic conditions when both are heavily phosphorylated. These results link vesicle docking, stacking of Golgi cisternae, and the disruption of both of these interactions during mitosis. PMID- 9346243 TI - Structure of the substrate binding domain of the thermosome, an archaeal group II chaperonin. AB - The crystal structure of the substrate binding domain of the thermosome, the archaeal group II chaperonin, has been determined at 2.3 A resolution. The core resembles the apical domain of GroEL but lacks the hydrophobic residues implied in binding of substrates to group I chaperonins. Rather, a large hydrophobic surface patch is found in a novel helix-turn-helix motif, which is characteristic of all group II chaperonins including the eukaryotic TRiC/CCT complex. Models of the holochaperonin, which are consistent with cryo electron microscopy data, suggest a dual role of this helical protrusion in substrate binding and controlling access to the central cavity independent of a GroES-like cochaperonin. PMID- 9346244 TI - Encapsidated conformation of bacteriophage T7 DNA. AB - The structural organization of encapsidated T7 DNA was investigated by cryo electron microscopy and image processing. A tail-deletion mutant was found to present two preferred views of phage heads: views along the axis through the capsid vertex where the connector protein resides and via which DNA is packaged; and side views perpendicular to this axis. The resulting images reveal striking patterns of concentric rings in axial views, and punctate arrays in side views. As corroborated by computer modeling, these data establish that the T7 chromosome is spooled around this axis in approximately six coaxial shells in a quasi crystalline packing, possibly guided by the core complex on the inner surface of the connector. PMID- 9346245 TI - A gene essential for de novo methylation and development in Ascobolus reveals a novel type of eukaryotic DNA methyltransferase structure. AB - Molecular mechanisms determining methylation patterns in eukaryotic genomes still remain unresolved. We have characterized, in Ascobolus, a gene for de novo methylation. This novel eukaryotic gene, masc1, encodes a protein that has all motifs of the catalytic domain of eukaryotic C5-DNA-methyltransferases but is unique in that it lacks a regulatory N-terminal domain. The disruption of masc1 has no effect on viability or methylation maintenance but prevents the de novo methylation of DNA repeats, which takes place after fertilization, through the methylation induced premeiotically (MIP) process. Crosses between parents harboring the masc1 disruption are arrested at an early stage of sexual reproduction, indicating that the activity of Masc1, the product of the gene, is crucial in this developmental process. PMID- 9346268 TI - Evaluation of threshold criteria for the nasal histamine challenge test in perennial allergic rhinitis. AB - Nasal reactivity to histamine was determined in patients with perennial allergic rhinitis and in control subjects. A histamine titration method delivered by a metered dose pump was used. Stuffiness, itching, and the number of sneezes were recorded, nasal secretions measured, and nasal airway resistance was recorded by active anterior rhinomanometry. Increased nasal reactivity to histamine was observed among rhinitic patients and inversely correlated with the severity of nasal symptoms. A 3-fold increase of post-saline nasal airway resistance (NAR) best differentiated the nasal responses to histamine in rhinitic patients from those in control subjects. A histamine dose of < or = 2.5 microg provoked a 3 fold increase in NAR, strongly suggesting moderate or severe symptomatic rhinitis in most cases. Nasal provocation techniques might be a useful tool for objectively assessing disease severity and response to treatment in perennial allergic rhinitis. PMID- 9346269 TI - Sulphasalazine therapy in chronic uveitis of children with chronic arthritis. AB - Four children with chronic arthritis (3 juvenile rheumatoid arthritis and 1 juvenile ankylosing spondylitis) and poorly controlled chronic uveitis, were given sulphasalazine (SASP) therapy for a mean period of 3.3 years. Three patients showed an excellent response, as evidenced by a reduction of inflammatory cells in the anterior chamber of the eyes and improvement of visual acuity. The response occurred after a mean of 7.7 weeks. These data suggested SASP therapy may have a role in the treatment of chronic anterior uveitis in children with chronic arthritis. PMID- 9346270 TI - Effect of histamine on lecithin content in broncho-alveolar lavage fluid of rat. AB - Deficiency of surfactant in alveoli leads to increased resistance to breathing. Histamine is a mediator in allergic respiratory diseases. Though the bronchoconstrictor effect of histamine is well recognised, histamine may have additional actions that contribute to pathogenesis in these diseases. The present study aimed to observe the effect of histamine on lecithin, a major component of alveolar surfactant. Lecithin content in broncho-alveolar lavage (BAL) fluid of healthy adult male rats was estimated by enzymatic method using Boehringer Mannheim kits. Lecithin content in these control animals was compared with that in three groups of healthy adult male rats following subcutaneous administration of 0.06 mg of histamine diphosphate at 10 minutes, 30 minutes and 60 minutes intervals, respectively. A significant reduction in lecithin levels in BAL fluid was observed up to one hour after administration of histamine. The results indicate a possible additional action of histamine in the pathogenesis of allergic respiratory diseases. PMID- 9346271 TI - Immunologic reactivity on one year follow-up of subjects without allergy to Hymenoptera stings. AB - We studied Hymenoptera stings in 72 pest-control operators without any previous systemic reactions to Hymenoptera stings, and investigated their venom-specific IgE levels in serial specimens collected over one year. At the initial evaluation, venom-specific IgE was present in 25 (34.7%) of 72 pest-control operators, and venom-specific IgE titer significantly decreased as the time interval from the last sting increased (p < 0.001). In most cases, venom-specific IgE disappeared less than 3 years after the last sting. On the other hand, the ratio of subjects with positive CAP for venom-specific IgE was significantly increased with an elevation of total serum IgE level (p < 0.001). After the one year follow-up, venom-specific IgE titer in the 25 subjects with positive CAP decreased significantly (p = 0.026). Total serum IgE level modified the decline significantly (p = 0.011), but the time interval from the last sting did not. In elevated total IgE level (>250 IU/ml), the decline of venom-specific IgE tended to be slow. PMID- 9346272 TI - High prevalence of antibody against hepatitis A virus in an institution for the mentally handicapped. AB - Hepatitis A virus infection constitutes a world-wide public health problem, predominantly in developing countries. Mentally handicapped children, due to their incapacity for looking after themselves, comprise one of the high risk groups for hepatitis A virus infection. The aim of the present study was to determine the prevalence of hepatitis A virus antibody (anti-HAV) among the children and adults in the Institute for the Mentally Handicapped located in Nonthaburi, Thailand. The prevalence of anti-HAV IgG antibody was 92%. Immunity acquired against HAV was shown to increase in direct proportion to the age. To prevent future outbreaks of hepatitis A, water supply, sanitary conditions and personal hygiene should be improved at this and similar institutions. Furthermore, persons new to the institution (patients and staff) should be screened for anti-HAV and vaccinated with hepatitis A vaccine if nonimmune. PMID- 9346273 TI - Immune status in congenital infections by TORCH agents in pregnant Thais. AB - A cross-sectional, sero-epidemiological survey of the prevalence of antibodies to TORCH agents during various stages of gestation revealed an overall rate of 13-15 percent having antibodies to Toxoplasma gondii; 85-87 percent, to rubella ; 79-81 percent, to herpes simplex virus (HSV); 100 percent, to cytomegalovirus (CMV); 82 86 percent, to human herpes virus type 6 (HHV-6); 1-2 percent, to hepatitis C virus (HCV). None of human T lymphotropic virus type I (HTLV-I) antibody was detected, and a prevalence of hepatitis B surface antigen (HBsAg) was 6 percent. Although a tendency was noted towards an increase of antibody detection to each TORCH agent as gestation progressed, a statistically significant increase in antibodies titer and specific IgM antibody was found with regard to CMV. These results suggest an increase in CMV infection or reactivation during pregnancy whereas an increase in the other TORCH infections was not obvious. PMID- 9346274 TI - Simplified, rapid diagnosis of respiratory syncytial virus from clinical specimens. AB - DOT ELISA was compared with RT-PCR and tissue culture to detect RSV from nasopharyngeal aspirates. DOT ELISA had diagnostic sensitivity and specificity of 65.62% and 93.92%, respectively. The results indicate that DOT ELISA can be used for screening detection of RSV from clinical specimens and is suitable for small laboratories in the provincial areas of developing countries. PMID- 9346275 TI - International clinical trials of HIV vaccines: II. phase I trial of an HIV-1 synthetic peptide vaccine evaluating an accelerated immunization schedule in Yunnan, China. AB - A Phase 1, double-blind, placebo controlled trial was conducted in Longchuan County, China, to evaluate the safety and immunogenicity of a prototype HIV-1 synthetic peptide vaccine in a target population at risk for HIV infection, and to establish the infrastructure for future large-scale HIV vaccine efficacy trials. Subjects were randomly assigned to receive 100 microg or 500 microg of vaccine or alum placebo, and were given three injections at an accelerated 0, 1, and 2 month schedule. The vaccine was well tolerated with no significant local or systemic reactions observed in any subjects. Fifty-five percent (100 microg dose) and 64% (500 microg dose) of subjects who received the vaccine produced binding antibody to the immunogen as determined by ELISA. However, HIV-1 (MN) neutralizing antibody was detected in only 23% (3/13) of subjects with detectable HIV-1 specific binding antibody. It was concluded that this prototype HIV-1 synthetic peptide vaccine was well tolerated, safe and immunogenic, and that a 0, 1, 2 month schedule was not as effective in stimulating HIV-1 specific neutralizing antibodies compared with previous trials utilizing a 0, 1, 6 month schedule. Finally, this trial demonstrated that well-designed HIV vaccine trials can be performed at this clinical trials site in Yunnan, China, and that this site should be considered for conducting larger safety, immunogenicity and efficacy trials of candidate HIV vaccines. PMID- 9346276 TI - Parasites elicited cross-reacting antibodies to Opisthorchis viverrini. AB - Two batches of crude antigens extracted from adult Opisthorchis viverrini worms were compared. One was derived from adult worms harvested from the livers of laboratory infected hamsters and another was obtained from worms sedimented from the faeces of opisthorchiasis patients following treatment with Praziquantel. SDS PAGE and Coomassie brilliant blue staining revealed that the two preparations had similar protein components of which the predominant ones were the 17-18 kDa doublet. The antigens were used in an indirect ELISA for the detection of antibodies against O. viverrini in the sera of four groups of patients, ie. patients with opisthorchiasis (group 1), patients with mixed infections of O. viverrini and other parasites (group 2), patients with other parasitic infections (group 3), and normal-heathy, parasite-free individuals (group 4). The sensitivity of the test was high (91-92%), regardless of the batch of the antigen used. However, its specificity was relatively low (70-80%). Cross-reaction was observed with patients infected with Paragonimus heterotremus, Schistosoma spp.; Taenia spp.; Trichinella spiralis; Strongyloides stercoralis; hookworms; Plasmodium spp.; hookworms and Plasmodium spp.; S. stercoralis, Blastocystis hominis and yeast; and hookworms, Ascaris lumbricoides, Trichuris trichiura and P. falciparum. Western blot analysis revealed that sera of patients infected with these heterologous organisms contained antibodies reactive to O. viverrini antigenic components ranging from Mr 15.5 to 144. PMID- 9346277 TI - GTP-binding-protein-coupled receptor kinases--two mechanistic models. AB - Six vertebrate protein kinases (G-protein-coupled receptor kinases; GRKs) that regulate the function of G-protein-coupled receptors (GPCRs) were recently cloned; several distinct properties set them apart from conventional second messenger regulated protein kinases. It appears that GRKs bind GPCR* through two separate sites: a high-affinity site, which involves intracellular loops of the activated receptor, and the lower-affinity site, encompassing the phosphorylation region. The high-affinity interaction may involve complementary structural elements of GRKs and GPCRs* rather than precise amino acid alignment, thus allowing broad and overlapping specificities of these kinases, in spite of differences in the sequences of GPCRs. In addition, GRK structures are modified by several posttranslational modifications, including phosphorylation, autophosphorylation, prenylation, carboxymethylation, and palmitoylation, probably affecting properties of these enzymes. While GRKs phosphorylate and inactivate receptor molecules which are engaged in G-protein activation, controversy surrounds whether GRKs might be activated and phosphorylate unstimulated GPCRs, leading to a desensitization of a larger population of the receptors. In this review, mechanistic aspects of GPCR* phosphorylation related to the distinct properties, regulation and modes of action of GRKs are described. PMID- 9346278 TI - Specificity of serine proteinase/serpin complex binding to very-low-density lipoprotein receptor and alpha2-macroglobulin receptor/low-density-lipoprotein receptor-related protein. AB - Very-low-density lipoprotein receptor (VLDLR) and alpha2-macroglobulin receptor/low-density-lipoprotein-receptor-related protein (alpha2MR/LRP) are multifunctional endocytosis receptors of the low-density lipoprotein receptor family. Both have been shown to mediate endocytosis and degradation of complex between plasminogen activators and type-1 plasminogen-activator inhibitor (PAI-1) by cultured cells. We have now studied the specificity of binding and endocytosis by VLDLR and alpha2MR/LRP among a variety of serine proteinase/serpin complexes, including various combinations of the serine proteinases urokinase-type and tissue-type plasminogen activators, plasmin, thrombin, human leukocyte elastase, cathepsin G, and plasma kallikrein with the serpins PAI-1, horse leukocyte elastase inhibitor, protein C inhibitor, C1-inhibitor, alpha2-antiplasmin, alpha1 proteinase inhibitor, alpha1-antichymotrypsin, protease nexin-1, heparin cofactor II, and antithrombin III. Binding was estimated with radiolabelled ligands in ligand blotting analysis and microtiter well assays. Endocytosis was estimated by measuring receptor-associated protein (RAP)-sensitive degradation of radiolabelled complexes by Chinese hamster ovary cells transfected with VLDLR cDNA and by COS-1 cells, which have a high endogenous expression of alpha2MR/LRP. We found that the receptors bind with high affinity to some, but not all, combinations of plasminogen activators and thrombin with PAI-1, protease nexin-1, protein C inhibitor, and antithrombin III, while complexes of many serine proteinases with their primary inhibitor, i.e. plasmin/alpha2-antiplasmin complex, do not bind, or bind with a very low affinity. Both the serine proteinase and the serpin moieties contribute to the binding specificity. The binding specificities of VLDLR and alpha2MR/LRP are overlapping, but not identical. The results suggest that VLDLR and alpha2MR/LRP have different biological functions by having different binding specificities as well as by being expressed by different cell types. PMID- 9346279 TI - Mycothiol-dependent formaldehyde dehydrogenase, a prokaryotic medium-chain dehydrogenase/reductase, phylogenetically links different eukaroytic alcohol dehydrogenases--primary structure, conformational modelling and functional correlations. AB - Prokaryotic mycothiol-dependent formaldehyde dehydrogenase has been structurally characterized by peptide analysis of the 360-residue protein chain and by molecular modelling and functional correlation with the conformational properties of zinc-containing alcohol dehydrogenases. The structure is found to be a divergent medium-chain dehydrogenase/reductase (MDR), at a phylogenetic position intermediate between the cluster of dimeric alcohol dehydrogenases of all classes (including the human forms), and several tetrameric reductases/dehydrogenases. Molecular modelling and functionally important residues suggest a fold of the mycothiol-dependent formaldehyde dehydrogenase related overall to that of MDR alcohol dehydrogenases, with the presence of the catalytic and structural zinc atoms, but otherwise much altered active-site relationships compatible with the different substrate specificity, and an altered loop structure compatible with differences in the quaternary structure. Residues typical of glutathione binding in class-III alcohol dehydrogenase are not present, consistent with that the mycothiol factor is not closely similar to glutathione. The molecular architecture is different from that of the 'constant' alcohol dehydrogenases (of class-III type) and the 'variable' alcohol dehydrogenases (of class-I and class II types), further supporting the unique structure of mycothiol-dependent formaldehyde dehydrogenase. Borders of internal chain-length differences between this and other MDR enzymes coincide in different combinations, supporting the concept of limited changes in loop regions within this whole family of proteins. PMID- 9346280 TI - Basic residues in the 74-83 and 191-198 segments of protein kinase CK2 catalytic subunit are implicated in negative but not in positive regulation by the beta subunit. AB - Protein kinase CK2 is a ubiquitous pleiotropic serine/threonine protein kinase whose holoenzyme is comprised of two catalytic (alpha and/or alpha') and two non catalytic, beta-subunits. The beta-subunit possesses antagonist functions that can be physically dissected by generating synthetic fragments encompassing its N terminal and C-terminal domains. Here we show that by mutating basic residues in the 74-77 and in the 191-198 regions of the alpha-subunit, the negative regulation by the beta-subunit and by its N-terminal synthetic fragment CK2beta (1-77), which is observable using calmodulin as a substrate for phosphorylation, is drastically reduced. In contrast, the positive regulation by a C-terminal, CK2beta-(155-215)-peptide is unaffected or even increased. Moreover, the basal activity of alpha mutants K74-77A, K79R80K83A, and R191R195K198A toward specific peptide substrates is stimulated by the beta-subunit many fold more than that of alpha wild type, while extrastimulation by beta mutant D55L56E57A, observable with alpha wild type, is abolished with these mutants. These data support the conclusion that down regulation by the acidic residues clustered in the N terminal moiety of beta is mediated by basic residues in the 74-83 and in the 191 198 sequences of the alpha-subunit. These are also implicated in substrate recognition consistent with the concept that the N-terminal acidic region of the beta subunit operates as a pseudosubstrate. In contrast, another CK2alpha mutant, V66A, is more sensitive to inhibition by either beta-subunit or its N-terminal, CK2beta-(1-77)-peptide, while its stimulation by the C-terminal peptide, CK2beta (155-215), is comparable to that of alpha wild type. These observations suggest an indirect role of Val66 in conferring to the alpha-subunit a conformation less sensitive to down regulation by beta-subunit. PMID- 9346281 TI - Specific alpha-galactosidase inhibitors, N-methylcalystegines--structure/activity relationships of calystegines from Lycium chinense. AB - An examination of the roots of Lycium chinense (Solanaceae) has resulted in the discovery of 14 calystegines, a cycloheptane bearing an amino group and three hydroxyl groups, and two polyhydroxylated piperidine alkaloids. Calystegines A7 and B5, in addition to the previously known calystegines A3, A5, A6, B1, B2, B3, B4, C1, C2 and N1, were isolated and determined as 1alpha,2beta,4alpha-trihydroxy nortropane and 1alpha,2alpha,4alpha,7alpha-tetrahydroxy-nort ropane, respectively. L. chinense also had two polyhydroxytropanes bearing a methyl group on the nitrogen atom, unlike the previously reported nortropane alkaloids. They were established as N-methylcalystegines B2 and C1, and their N-methyl groups were found to be axially oriented from NOE experiments. 1Beta-amino 3beta,4beta,5alpha-trihydroxycyclohepta ne was also present in L. chinense and may be a biosynthetic precursor of the calystegines that occur in this plant. Two polyhydroxypiperidine alkaloids, fagomine and 6-deoxyfagomine, were isolated. Calystegine B2 is a potent competitive inhibitor of almond beta-glucosidase (Ki = 1.9 microM) and coffee bean alpha-galactosidase (Ki = 0.86 microM), while N methylcalystegine B2 was a more potent competitive inhibitor of the latter enzyme (Ki = 0.47 microM) than the parent compound but showed a marked lack of inhibitory activities towards most other glycosidases. Since this compound is a very specific inhibitor of alpha-galactosidase and inhibits rat liver lysosomal alpha-galactosidase with a Ki of 1.8 microM, it may provide a useful experimental model for the lysosomal storage disorder, Fabry's disease. The addition of a hydroxyl group at C6exo, as in calystegines B1 and C1, enhances the inhibitory potential towards beta-glucosidase and beta-galactosidase but markedly lowers or abolishes inhibition towards alpha-galactosidase. Hence, the N-methylation of calystegine C1 did not enhance its inhibition of alpha-galactosidase. The chemical N-methylation of calystegines A3 and B4 markedly enhanced inhibition of coffee bean alpha-galactosidase, with Ki values of 5.2 microM and 36 microM, respectively, but almost eliminated their inhibitory potential towards beta glucosidase and trehalase, respectively. Thus, methylation of the nitrogen atom significantly altered the specificity of the inhibitors. PMID- 9346282 TI - Proteinase A, a storage-globulin-degrading endopeptidase of vetch (Vicia sativa L.) seeds, is not involved in early steps of storage-protein mobilization. AB - Proteinase A is a papain-like cysteine endopeptidase of vetch (Vicia sativa L.) which was assumed to initiate storage-globulin breakdown just after the onset of seed germination. This enzyme was purified from cotyledons of vetch seedlings. On gelatin-containg SDS gels, active proteinase A migrated with an apparent molecular mass of 21 kDa, whereas after heat denaturation its molecular size on SDS/PAGE was 29 kDa. Although proteinase A is capable of hydrolyzing storage globulins in vitro it could not be localized in the protein-body fraction of cotyledons from germinating seeds. cDNA clones encoding proteinase A precursor have been obtained by PCR. The precursor is composed of an N-terminal signal sequence followed by a propeptide, the region encoding mature proteinase A, and a C-terminal KDEL sequence. Mature proteinase A with a derived molecular mass of 25,244 Da does not have the KDEL sequence. The derived amino acid sequence of the proteinase A precursor is 78.2% identical to sulfhydryl-endopeptidase (SH-EP), a cysteine endopeptidase from germinating Vigna mungo seedlings. Northern blot analysis indicated that proteinase A mRNA appears de novo in cotyledons of 1-day germinated vetch seeds, where its amount increases up to day 6. No proteinase A mRNA was detected in other vetch organs, not even in the embryo axis, which contains stored globulins. By means of antibodies raised against the purified and against recombinantly produced proteinase A, the 29-kDa bands of mature proteinase A were detected in cotyledon extracts of 6-day-germinated seeds when globulin degradation has already far proceeded. The reported data do not agree with the proposed triggering role of proteinase A in storage-globulin breakdown during germination. PMID- 9346283 TI - Structure of HOE/BAY 793 complexed to human immunodeficiency virus (HIV-1) protease in two different crystal forms--structure/function relationship and influence of crystal packing. AB - Human immunodeficiency virus 1 (HIV-1) protease is a prime target in the search for drugs to combat the AIDS virus. The enzyme functions as a C2-symmetric dimer, cleaving the gag and gag-pol viral polyproteins at distinct sites. The possession of a twofold axis passing through the active site, has led to the design of C2 symmetrical inhibitors in the form of substrate-based transition-state analogs. One of the most active compounds of this class of inhibitors is HOE/BAY 793, which contains a vicinal diol central unit [Budt, K.-H., Hansen, J., Knolle, J., Meichsner, C., Paessens, A., Ruppert, D. & Stowasser, B. & Winkler, I. (1990) European Patent application EP0428,849; Budt, K.-H., Hansen, J., Knolle, J., Meichsner, C., Ruppert, D., Paessens, A. & Stowasser B. (1993) IXth International Conference on AIDS; Budt, K.-H., Peyman, A., Hansen, J., Knolle, J., Meichsner, C., Paessens, A., Ruppert, D. & Stowasser, B. (1995) Bioorg. Med. Chem. 3, 559 571.] The structure of this inhibitor bound to HIV-1 protease, in two different crystal forms, has been solved at 0.24-nm resolution using X-ray crystallography. In both forms, the details of the inhibitor-protease interactions revealed an overall asymmetric binding mode, especially between the central diol unit and the active-site aspartates. The main binding interactions comprise several specific H bonds and hydrophobic contacts, which rationalize many of the characteristics of the structure/activity relationship in the class of vicinal diol inhibitors. In a general analysis of the mobility of the flap regions, which cover the active site and participate directly in binding, using our structures and the HIV protease models present in the Brookhaven databank, we found that in most structures the flexibility of the flaps is limited by local crystal contacts. However, in one of the structures presented here, no significant crystal contacts to the flap regions were present, and as a result the flexibility of the inhibitor bound flaps increased significantly. This suggests that the mobility and conformational flexibility of the flap residues are important in the functioning of HIV-1 protease, and must be considered in the future design of drugs against HIV protease and in structure-based drug design in general. PMID- 9346284 TI - Multiheme cytochromes from the sulfur-reducing bacterium Desulfuromonas acetoxidans. AB - Two new multiheme cytochromes were isolated from the anaerobic sulfur reducing bacterium Desulfuromonas acetoxidans. They have monomeric molecular masses of 50 and 65 kDa and contain six and eight hemes, respectively. Visible and EPR spectroscopies, in the as-isolated (oxidised) cytochromes, show the presence of only low-spin hemes in the 50-kDa cytochrome, and of high-spin and low-spin hemes in the 65-kDa cytochrome. The EPR spectra of the native 65-kDa cytochrome indicate multiple heme-heme interactions, including integer-spin systems as judged by parallel-mode EPR. The 50-kDa cytochrome has a complex redox pattern, as shown by EPR redox titrations, and contains one heme with unusual characteristics. Both cytochromes cover an extremely wide range of reduction potentials, which go from +100 mV to -375 mV for the 50-kDa cytochrome, and +185 mV to -235 mV for the 65-kDa cytochrome. The two cytochromes were tested for hydroxylamine oxidoreductase activity and polysulfide reductase activity, but neither displayed any activity. In contrast, it was found for the first time that the previously characterised cytochrome c551.5, from the same bacterium is very active in the reduction of polysulfide, which suggests that it acts as a terminal reductase in D. acetoxidans. PMID- 9346285 TI - The ordered phosphorylation of cardiac troponin I by the cAMP-dependent protein kinase--structural consequences and functional implications. AB - The pattern of phosphorylation of adjacent serine residues in several peptides based on the N-terminal region of human cardiac troponin I has been analysed by PAGE and 1H NMR spectroscopy to identify the products. With cAMP-dependent protein kinase, Ser24 is rapidly phosphorylated, and subsequent much slower phosphorylation of Ser23 occurs only after phosphorylation of Ser24 is almost complete. Monophosphorylation of the peptide at Ser23 was not detected at any time. On replacement of Arg22 with Ala or Met the sole phosphorylation target was Ser23, phosphorylation being considerably slower than for Ser24 in the wild-type peptide, while diphosphorylation could not be detected after prolonged incubation. The results emphasise the importance of the N-terminal sequence RRRSS for the function of cardiac troponin I and imply that in human cardiac muscle unstimulated by adrenaline, troponin I is phosphorylated on Ser24. Comparative two-dimensional NOESY data indicate that in the diphosphorylated form at physiological pH values, specific structural constraints are imposed on the N terminal peptide region. These constraints result in the effective screening of the two phosphate groups from each other by the arginine residues N-terminal to the serine pair and stabilisation of the structure in the region of residues 25 29, which is adjacent to a site of interaction between troponin I and troponin C. These conformational changes presumably underlie the decrease in calcium sensitivity of the myofibrillar ATPase that occurs after adrenaline intervention. PMID- 9346286 TI - Studies on the NusB protein of Escherichia coli--expression and determination of secondary-structure elements by multinuclear NMR spectroscopy. AB - The product of the nusB gene of Escherichia coli modulates the efficiency of transcription termination at nut (N utilization) sites of various bacterial and bacteriophage lambda genes. Similar control mechanisms operate in eukaryotic viruses (e.g. human immunodeficiency virus). A recombinant strain of E. coli producing relatively large amounts of NusB protein (about 10% of cell protein) was constructed. The protein could be purified with high yield by anion-exchange chromatography followed by gel-permeation chromatography. The protein is a monomer of 15.6 kDa as shown by analytical ultracentrifugation. Structural studies were performed using protein samples labelled with 15N, 13C and 2H in various combinations. Heteronuclear three-dimensional triple-resonance NMR experiments combined with a semi-automatic assignment procedure yielded the sequential assignment of the 1H, 13C and 15N backbone resonances. Based on experimentally derived scalar couplings, chemical-shift values, amide-exchange data, and a semiquantitative interpretation of NOE data, the secondary structure of NusB has classified as alpha helical, comprising seven alpha helices. PMID- 9346287 TI - Chemical, spectroscopic and structural investigation of the substrate-binding site in ascorbate peroxidase. AB - The interaction of recombinant ascorbate peroxidase (APX) with its physiological substrate, ascorbate, has been studied by electronic and NMR spectroscopies, and by phenylhydrazine-modification experiments. The binding interaction for the cyanide-bound derivative (APX-CN) is consistent with a 1:1 stoichiometry and is characterised by an equilibrium dissociation binding constant. Kd, of 11.6 +/- 0.4 microM (pH 7.002, mu = 0.10 M, 25.0 degrees C). Individual distances between the non-exchangeable substrate protons of APX-CN and the haem iron were determined by paramagnetic-relaxation NMR measurements, and the data indicate that the ascorbate binds 0.90-1.12 nm from the haem iron. The reaction of ferric APX with the suicide substrate phenylhydrazine yields predominantly (60%) a covalent haem adduct which is modified at the C20 carbon, indicating that substrate binding and oxidation is close to the exposed C20 position of the haem, as observed for other classical peroxidases. Molecular-modelling studies, using the NNM-derived distance restraints in conjunction with the crystal structure of the enzyme [Patterson, W. R. & Poulos, T. L. (1995) Biochemistry 34, 4331-4341], are consistent with binding of the substrate close to the C20 position and a possible functional role for alanine 134 (proline in other class-III peroxidases) is implicated. PMID- 9346288 TI - Identification of a putative histidine base and of a non-protein nitrogen ligand in the active site of Fe-hydrogenases by one-dimensional and two-dimensional electron spin-echo envelope-modulation spectroscopy. AB - The active H-cluster of the Fe-hydrogenases from Megasphaera elsdenii and Desulfovibrio vulgaris (strain Hildenborough) has been investigated with one- and two-dimensional pulsed EPR spectroscopy. In both complexes the coordination of a nitrogen-containing ligand was found. The unusual quadrupole interaction parameters (D. vulgaris: quadrupole coupling constant, K = 1.20 MHz, asymmetry parameter eta = 0.32, M. elsdenii: K = 1.23 MHz, eta = 0.25) indicate a non protein type of nitrogen and are consistent with cyanide as ligand to the H cluster. The additional interactions measured on the EPR signal of the inactivated H-cluster in D. vulgaris hydrogenase are consistent with an imidazole interaction similar to that found in Rieske-type iron-sulfur clusters. Since a His residue near the putative H-cluster binding motif of Cys residues, His371, is the only conserved His in Fe-hydrogenases, it is a likely candidate for the base that accepts the proton in the heterolytic cleavage of molecular hydrogen. The inactivation of the enzyme is accompanied by direct binding of the imidazole ring to the H-cluster. PMID- 9346289 TI - Site-directed mutagenesis and halophilicity of dihydrolipoamide dehydrogenase from the halophilic archaeon, Haloferax volcanii. AB - A homology-modelled structure of dihydrolipoamide dehydrogenase from the halophilic archaeon, Haloferax volcanii, has been generated using the crystal structure of the enzyme from Pseudomonas fluorescens. Analysis of the halophilic enzyme structure identified a potential K+-binding site comprising four co ordinated glutamate residues (E423 and E426 from each monomer) at the subunit interface of the dimeric protein. Whilst E426 is conserved throughout non halophilic dihydrolipoamide dehydrogenases, E423 is only present in the halophilic enzyme. Four site-directed mutations of the Haloferax dihydrolipoamide dehydrogenase have been made (E423D, E423Q, E423S, and E423A) and the recombinant mutants expressed and characterised. From an analysis of their kinetic properties, salt-dependent activities and thermal stabilities, it is concluded that this site has an important influence on the halophilicity of the enzyme. The findings support the view that the arrangement and interaction of the negatively charged amino acids are as important as the total net charge in determining the adaptation of proteins to high salt concentrations. PMID- 9346290 TI - Activation of progelatinase B (proMMP-9) by active collagenase-3 (MMP-13). AB - Human progelatinase B was activated by collagenase-3 in a time-dependent fashion. Activation proceeded through an intermediate form of Mr 86,000 to the final active form of Mr 82,000. N-terminal amino acid sequence determination demonstrated that the Glu40-Met41 peptide bond was initially hydrolysed followed by cleavage of the Arg87-Phe88 peptide bond releasing the rest of the propeptide domain which was accompanied by the achievement of maximal enzymatic activity as revealed using a quenched fluorescent substrate. Kinetic analysis of activation revealed that the rates were dependent on the concentration of the proenzyme as well as active collagenase-3. Active gelatinase B did not contribute to the activation rate of the proenzyme initiated by collagenase-3 and our results indicate that progelatinase B activation proceeds via bimolecular cleavage with collagenase-3 involving sequential cleavage of the propeptide in two steps. The activation rates were not dependent on C-terminal domain interactions between progelatinase B and collagenase-3, as assessed using wild-type and C-terminal deletion mutants of both enzymes. Since elevated levels of both gelatinase B and collagenase-3 have been observed in arthritis and breast cancer pathology these enzymes may well form a proteolytic cascade in these diseases which allows rapid turnover of the extracellular matrix. PMID- 9346291 TI - Study of fatty acid specificity of sunflower phospholipase D using detergent/phospholipid micelles. AB - The fatty acid specificity of phospholipase D purified from germinating sunflower seeds was studied using mixed micelles with variable detergent/phospholipid ratios. The main advantage of this approach is that since the substrate is integrated in the detergent micelles, comparisons can be made between the kinetic constants of a wide range of phosphatidylcholine (PtdCho) compounds with various fatty acid contents. Phospholipase D is subject to interfacial activation as it is most active on water-insoluble substrates. It is not active on sphingomyelin and only slightly on lysophosphatidylcholine. By fitting the curves based on the experimental kinetic data, the interfacial dissociation constant of phospholipase D, the maximum hydrolysis rate Vm and the kinetic constant Km(B), were determined with the micellar substrate. The specificity of various substrates was examined by comparing the Vm/Km(B) values, and it was noted that sunflower phospholipase D is most active on medium-chain fatty PtdCho compounds. With long-chain natural phospholipids, the specificity of phospholipase D was slightly dependent on the level of fatty acid unsaturation. The pure enzyme was able to hydrolyse the sunflower phospholipids present in mixed detergent micelles but not the phospholipids integrated in the natural sunflower oil body structure. We concluded, however, that during the germination of sunflower seeds, phospholipase D might be involved in the degradation of oil bodies, since other factors present in crude seed extracts may make phospholipids accessible to the enzyme. PMID- 9346292 TI - Mossbauer study of 4-hydroxybutyryl-CoA dehydratase--probing the role of an iron sulfur cluster in an overall non-redox reaction. AB - 4-Hydroxybutyryl-CoA dehydratase from Clostridium aminobutyricum catalyzes the dehydration of 4-hydroxybutyryl-CoA to crotonyl-CoA. Although dehydration is an overall non-redox reaction, the enzyme contains FAD and Fe-S clusters. Previous work has shown that the Fe-S clusters are difficult to reduce and therefore unlikely to be redox-active in catalysis. Here, Mossbauer spectroscopy has been used to characterise the Fe-S clusters in active as well as in air-inactivated enzyme. In zero magnetic field at 80 K and 4.2 K, the spectra of active dehydratase consisted mainly of one species (95%) with quadrupole splitting, deltaE(Q) = 1.00 mm s(-1) and isomer shift, delta = 0.43 mm s(-1). Magnetically perturbed Mossbauer spectra indicated a spin of zero. In the presence of 6 mM crotonyl-CoA, the spectra remained unchanged. Taken together, the data show that there are [4Fe-4S]2+ in the enzyme, most probably two clusters/homotetramer, that the four iron atoms in each cluster are coordinated in an identical fashion, and that there is no direct interaction with substrates. We therefore infer that the Fe-S clusters serve a structural rather than a catalytic role in 4-hydroxybutyryl CoA dehydratase. In air-inactivated enzyme (10% residual activity), a new doublet appeared (58%) with deltaE(Q) = 0.72 mm s(-1), delta = 0.32 mm s(-1) and S = 0. The assignment of this subspectrum to [3Fe-4S]+ clusters, based on the typical Mossbauer parameters, is contradicted by the finding of spin zero for the species. One possible explanation could be spin-coupling of two [3Fe-4S]+ clusters in close proximity. PMID- 9346293 TI - Isolation and characterization of D-threonine aldolase, a pyridoxal-5'-phosphate dependent enzyme from Arthrobacter sp. DK-38. AB - D-Threonine aldolase is an enzyme that catalyzes the cleavage of D-threonine into glycine and acetaldehyde. Its activity was found in several genera of bacteria such as Arthrobacter, Alcaligenes, Xanthomonas, and Pseudomonas, but not in yeasts or fungi. The enzyme was purified to homogeneity from one strain, Arthrobacter sp. DK-38. The enzyme appeared to consist of a single polypeptide chain with an apparent molecular mass of 51 kDa. This enzyme, as well as L threonine aldolase, requires pyridoxal 5'-phosphate (pyridoxal-P) as a coenzyme. Unlike other pyridoxal-P enzymes, D-threonine aldolase also requires a divalent cation such as Co2+, Ni2+, Mn2+, or Mg2+ for its catalytic activity. The enzyme completely lost its activity in the absence of either pyridoxal-P or a divalent cation. A divalent cation was also essential for the thermal stability of the enzyme. The metal-free enzyme tends to become thermally unstable, resulting in the irreversible loss of its catalytic activity. The enzyme is strictly D specific for the alpha-position, whereas it cannot distinguish between threo and erythro forms at the beta-position. Thus, D-threonine and D-allothreonine act as substrates of the enzyme, but their kinetic parameters are different; the Km and Vmax values are 3.81 mM and 38.8 micromol x min(-1) x mg(-1) toward D-threonine, and 14.0 mM and 102 micromol x min(-1) x mg(-1) toward D-allothreonine. respectively. The aldolase reaction is reversible, and the enzyme is therefore able to produce nearly equimolar amounts of D-threonine and D-allothreonine through C-C bond formation between glycine and acetaldehyde. The enzyme also acts, in the same manner, on several other D-beta-hydroxy-alpha-amino acids, including D-beta-phenylserine, D-beta-hydroxy-alpha-aminovaleric acid, D-beta-3,4 dihydroxyphenylserine, and D-beta-3,4-methylenedioxyphenylserine. PMID- 9346294 TI - Sphingosylphosphorylcholine activates an amiloride-insensitive Na+-H+-exchange mechanism in GH4C1 cells. AB - The effect of sphingosylphosphorylcholine (SphPCho) on the intracellular pH (pHi) in GH4C1 cells was investigated. SphPCho evoked a very slow increase in basal pHi. In cells acidified with nigericin, SphPCho induced a rapid alkalinization of the cells. The effect was inhibited in a Na+-free buffer solution, but was insensitive to ethylisopropyl amiloride, a potent inhibitor of Na+-H+ exchangers (NHE). Reverse transcription and PCR showed that the predominant isoform of the antiport expressed in GH4C1 cells is NHE-1. The rate of alkalinization after stimulation with propionate, and after addition of Na+ to cells acidified with NH4Cl, was enhanced in cells treated with SphPCho. The initial rate of alkalinization after addition of Na+ to acidified cells treated with SphPCho gave an apparent Km value of 15 +/- 2 mM for Na+. The Vmax value was 9 +/- 2 mM H+/min. The effect was insensitive to ouabain, staurosporine and bafilomycin A. However, the SphPCho-evoked alkalinization was abolished in cells treated with 2 deoxy-D-glucose. The effect was not due to the charge of the molecule, as stearylamine increased pHi in Na+-containing and Na+-free buffer. The results show that SphPCho may activate Na+-H+ exchange, and that this effect is mediated via an amiloride-insensitive exchange mechanism. PMID- 9346295 TI - Metabolism of 14C-labelled 5-nitro-1,2,4-triazol-3-one by rat liver microsomes- evidence for the participation of cytochrome P-450. AB - In the present study, we synthesized 14C-labelled 5-nitro-1,2,4-triazol-3-one (NTO) and investigated its hepatic metabolism by dexamethasone-induced murine hepatic microsomes. Under the nitrogen atmosphere, 5-amino-1,2,4-triazol-3-one was the only detected metabolite of NTO. The microsomal nitroreductase activity was dependent on NADPH, totally inhibited by carbon monoxide and partially inhibited by oxygen. In aerobic conditions, beside a low amount of amine, the major metabolite formed is the 5-hydroxy-triazolone, urazole. This compound resulted from the oxidative denitrification of NTO, which produced equivalent amount of nitrite. This reaction, like the nitroreductase activity, was dependent on NADPH and totally inhibited by carbon monoxide. Both nitroreduction and oxidative denitrification were inhibited by imidazole-related inhibitors: miconazole and methimazole, and to a less extent by N-octylamine. The microsomal denitrification was induced by the treatment of rats with dexamethasone and phenobarbital. The microsomal reductase activity is present in untreated rat microsomes, and recovered with various inducers. The results of this study indicate the role played by cytochrome P-450 in the metabolism of NTO, supported by its transformation with reconstituted cytochrome P-450 systems. PMID- 9346296 TI - Characteristics of protein-kinase-C- and ADP-ribosylation-factor-stimulated phospholipase D activities in human embryonic kidney cells. AB - Phospholipase D (PLD) activity in human embryonic kidney (HEK) cells is stimulated by phorbol-ester-activated protein kinase C (PKC) and by membrane receptors, the latter apparently acting via the GTP-binding proteins, ADP ribosylation factor (ARF) and Rho. In the present study, performed in cell-free preparations, we have characterized and compared the regulation of HEK cell PLD activity by the stable GTP analogue, guanosine 5'-O-[gamma-thio]triphosphate (GTP[S]), and the phorbol ester, phorbol 12-myristate 13-acetate (PMA). In digitonin-permeabilized HEK cells, prelabeled with [3H]oleic acid, GTP[S] and PMA caused an approximately threefold concentration-dependent increase in the formation of [3H]phosphatidylethanol, measured in the presence of ethanol. Neomycin, which is known to complex with the PLD cofactor, phosphatidylinositol 4,5-bisphosphate, decreased basal and GTP[S]- or PMA-stimulated PLD activities with similar sensitivity. GDP and its analogue, guanosine 5'-O-[beta thio]diphosphate, inhibited the stimulatory effect of GTP[S], whereas the PMA response was prevented by the nonselective PKC inhibitor, staurosporine, but not vice versa. PLD stimulation by GTP[S], but not by PMA, was markedly reduced upon cytosol depletion and reconstituted by purified recombinant ARF1. In HEK cell membranes, addition of purified recombinant ARNO, a guanine-nucleotide-exchange factor for ARF1. potentiated the GTP[S]-stimulated PLD activity. PLD stimulation by PMA in HEK cell membranes required MgATP and was largely prevented by the selective PKC inhibitors Goe 6976 and bisindolylmaleimide I. Immunoblot analysis demonstrated that both conventional PKC (alpha, beta, gamma) and atypical PKC isozymes (zeta, tau) were present in HEK cell membranes. The results indicate that phorbol ester stimulation of PLD activity in HEK cells apparently occurs by a phosphorylation-dependent mechanism involving membrane-associated PKC isozymes but not ARF proteins, the major targets of GTP[S]' action. PMID- 9346297 TI - Differential expression of the vegetative and spore-bound hydrophobins of Trichoderma reesei--cloning and characterization of the hfb2 gene. AB - The hfb2 gene encoding the hydrophobin HFBII of the filamentous fungus Trichoderma reesei was isolated by heterologous hybridization using the vegetative hydrophobin I, hfb1, gene of T. reesei as a probe. The hfb2 gene codes for a typical fungal secreted hydrophobin of 71 amino acids containing eight cysteine residues. The amino acid similarity towards HFBI is 69%. The HFBII protein was isolated from the fungal spores by extraction with trifluoroacetic acid/acetonitrile solution, and by bubbling from the lactose-based culture medium. Expression of the hfb1 and hfb2 genes is divergent. hfb1 expression was only observed in vegetative cultures on glucose-containing and sorbitol containing media. It was not expressed on media containing complex plant polysaccharides, cellulose, xylan, cellobiose or lactose, whereas hfb2 was highly expressed in vegetative cultures on these media. Expression of hfb2 was also strongly induced by N and C starvation, by light and in conidiating cultures. PMID- 9346298 TI - A kinetic study of triple-helix formation at a critical R x Y sequence of the murine c-Ki-ras promoter by (A,G)- and (G,T) oligonucleotides. AB - The kinetics of triplex formation between the oligonucleotides d(AGGGAGG GAGGAAGGGAGGG) (20AG), d(TGGGTGGGTGGTTGGGTGGG) (20GT) and a 29-bp polypurine polypyrimidine sequence located in the c-Ki-ras promoter (D) was studied by electrophoretic experiments in 50 mM Tris/acetate, pH 7.4, 50 mM NaCl, 5 mM MgCl2. Rates of triplex formation were determined at three different temperatures (20 degrees C, 37 degrees C and 45 degrees C), under pseudo-first order conditions obtained by using the triplex-forming oligonucleotide (TFO) 500-fold in excess over the target duplex (5 nM). Measurements at TFO/target ratios of 20 and 100 were also carried out. At 37 degrees C the pseudo first-order constants, k(obs), were 18.9 x 10(-5) s(-1) for 20AG and 13.0 x 10(-5) s(-1) for 20GT, yielding association half-lives of 1 h and 1.5 h, respectively. Second-order association constants were found to be in the order of 10(2) M(-1) s(-1): these are slightly lower if compared with those measured for triplex formation by polypyrimidine (C,T) oligonucleotides (10(3) M(-1) s(-1)) [Maher, L. J., Dervan, P. B. & Wolf, B. J. (1990) Biochemistry 29, 8820-8826; Xodo, L. E. (1995) Eur. J. Biochem. 228, 918-926; Bates, P. J., Dosanjh, H. S., Jenkins, T. C., Laughton, C. A. & Neidle, S. (1995) Nucleic Acids Res. 23, 3627-3632] but dramatically lower when compared with duplex recombination from complementary strands (10(6) M(-1) s(-1)) [Craig, M. E., Crothers, D. M. & Doty, P. (1971) J. Mol. Biol. 62, 383 401; Porschke, D. & Eigen, M. (1971) J. Mol. Biol. 62, 361-381]. Dissociation rate constants, k(-1), were indirectly obtained from equilibrium constants (Kd) and found to be, at 37 degrees C, 6.7 x 10(-7) s(-1) and 5.4 x 10(-6) s(-1) for 20AG and 20GT, respectively. From the rate constants obtained at 20 degrees C, 37 degrees C and 45 degrees C we estimated activation energies of triplex formation between D plus 20AG and D plus 20GT of respectively 134 +/- 29 and 88 +/- 21 kJ/mol. Moreover, the activation energies for the reaction of triplex dissociation were 385 +/- 50 kJ/mol for 20AG and 330 +/- 42 kJ/mol for 20GT. Decreasing the TFO/target ratio from 500 to 100 or 20, we observed a concomitant decrease of the association rate, in keeping with the finding that triplex formation occurs through a bimolecular process. We found that the effect of salt on triplex formation is rather complex, as, the addition of 2 mM spermidine boosted the binding rate of 20GT, but slightly reduced that of 20AG; the increase of NaCl from 50 mM to 100 mM or 150 mM decreased the rate of triplex formation. Finally, the biological implications of the kinetic behaviour exhibited by the two triplex-forming oligonucleotides specific for the c-Ki-ras promoter are discussed. PMID- 9346299 TI - Mutational analysis of a surface area that is critical for the thermal stability of thermolysin-like proteases. AB - Site-directed mutagenesis was used to assess the contribution of individual residues and a bound calcium in the 55-69 region of the thermolysin-like protease of Bacillus stearothermophilus (TLP-ste) to thermal stability. The importance of the 55-69 region was reflected by finding that almost all mutations had drastic effects on stability. These effects (both stabilizing and destabilizing) were obtained by mutations affecting main chain flexibility, as well as by mutations affecting the interaction between the 55-69 region and the rest of the protease molecule. The calcium-dependency of stability could be largely abolished by mutating one of its ligands (Asp57 or Asp59). In the case of the Asp57-->Ser mutation, the accompanying loss in stability was modest compared with the effects of other destabilizing mutations or the effects of (combinations of) stabilizing mutations. The detailed knowledge of the stability-determining region of TLP-ste permits effective rational design of stabilizing mutations, which, presumably, are also useful for related TLP such as thermolysin. This is demonstrated by the successful design of a stabilizing salt bridge involving residues 65 and 11. PMID- 9346300 TI - Phosphorylation mutants of p53 show differential complex formation with putative dehydrogenase Tms1 of fission yeast. AB - The yeast tms1 gene was originally identified as a multi-copy suppressor of a lethal growth arrest caused by expression of a tumour mutant cDNA of p53 in fission yeast. The tms1 gene product (Tms1) was found to form stable complexes with p53 in yeast and in vitro; using purified recombinant proteins, the interaction was mapped to the C-terminal region of p53. This part is known to be modified by several protein kinases resulting in a transition of p53 from a latent to an activated state capable of transactivating various cellular genes involved in growth suppression or apoptosis. Since there is evidence for an evolutionary conservation of a Tms1-related protein in mammals, the effect of the phosphorylation status of the C-terminus of p53 on Tms1/p53 complex formation in vitro has been investigated. Whereas mutants changing the cdc2 phosphorylation site at position 315 of human p53 had only little effect on Tms1/p53 complex formation, we found that mutants involving the protein kinase CK2 site at position 392 showed a significantly decreased relative affinity for the Tms1 protein. The same result was obtained by using a C-terminal fragment of p53 which was phosphorylated by purified protein kinase CK2, suggesting that the complex formation of p53 with cellular C-terminal binding proteins like Tms1 impairs regulation by phosphorylation. PMID- 9346301 TI - The primary structure of the split-Soret cytochrome c from Desulfovibrio desulfuricans ATCC 27774 reveals an unusual type of diheme cytochrome c. AB - The complete amino acid sequence of the unusual diheme split-Soret cytochrome c from the sulphate-reducing Desulfovibrio desulfuricans strain ATCC 27774 has been determined using classical chemical sequencing techniques and mass spectrometry. The 247-residue sequence shows almost no similarity with any other known diheme cytochrome c, but the heme-binding site of the protein is similar to that of the cytochromes c3 from the sulphate reducers. The cytochrome-c-like domain of the protein covers only the C-terminal part of the molecule, and there is evidence for at least one more domain containing four cysteine residues, which might bind another cofactor, possibly a non-heme iron-containing cluster. This domain is similar to a sequence fragment of the genome of Archaeoglobus fulgidus, which confirms the high conservation of the genes involved in sulfate reduction. PMID- 9346302 TI - Lipoxygenase-2 oxygenates storage lipids in embryos of germinating barley. AB - Besides the pre-existing lipoxygenase (LOX-1) present in quiescent grains, a new lipoxygenase (LOX-2) is induced in embryos of germinating barley [Holtman, W. L., Van Duijn, G., Sedee, N. J. A. & Douma, A. C. (1996) Plant Physiol. 111, 569 576]. The fact that LOX-1 and LOX-2 form different products after incubation with linoleic acid, the (9S)- and (13S)-hydroperoxides, respectively [Van Aarle, P. G. M., De Barse, M. M. J., Veldink, G. A. & Vliegenthart, J. F. G. (1991) FEBS Lett. 280, 159-162; Doderer, A., Kokkelink, I., Van der Veen, S., Valk, B. E., Schram, A. W. & Douma, A. C. (1992) Biochim. Biophys. Acta 1120, 97-104], and differ in temporal expression, suggests different physiological functions for the two isoenzymes at the onset of germination. We aimed to obtain more information about these functions by studying the substrate and product specificities of both isoenzymes. Analyses of the products formed from linoleic acid confirmed that LOX 1 oxygenated at C9, and LOX-2 at C13. When testing more complex substrates, it was found that both LOX-1 and LOX-2 were capable of metabolizing esterified fatty acids. Km values from both isoenzymes for free fatty acids were much lower than for esterified fatty acids (7-35-fold for LOX-1 versus 2-8-fold for LOX-2). Interestingly, LOX-1 showed significantly higher Km values for esterified fatty acids than did LOX-2. This was reflected by analyses of the products formed from di- and tri-linoleoylglycerol; LOX-2 formed higher amounts of oxygenated polyunsaturated fatty acids within the esterified lipids than did LOX-1, with a corresponding larger extent of oxygenation. In order to identify potential endogenous substrates, we analyzed free and esterified lipids in total lipid extracts from barley after different periods of germination for LOX-derived products. The results indicated that esterified fatty acids were preferentially metabolized by LOX-2 activity. Analysis of the positional specificity within the lipids after alkaline hydrolysis revealed that only (13S)-hydroxy derivatives were formed, indicating the in vivo action of LOX-2. These data show that LOX-2 is capable of oxygenating storage lipids and suggest that during the onset of germination LOX-2 may be involved in oxygenation of esterified polyunsaturated fatty acids in barley seeds. We suggest that the oxygenation of these lipids precedes the onset of their catabolism and that the degradation product, (9Z,11E,13S)-13-hydroxy-octadecadienoic acid, serves as an endogenous substrate for beta-oxidation and therefore as a carbon source for the growing barley embryo. PMID- 9346303 TI - Structural requirements of phospholipase C delta1 for regulation by spermine, sphingosine and sphingomyelin. AB - We studied the relationship between sphingomyelin, calcium, spermine and sphingosine in regulation of phospholipase C (PLC) delta1 activity. Inhibition of PLC delta1 by sphingomyelin was promoted by spermine and Ca2+ and was partially abolished by sphingosine. The effect of sphingosine and spermine entirely depended on Ca2+. In the absence of Ca2+, no effect of these substances on PLC delta1 activity was observed. Using deletion mutants and active fragments of PLC delta1 generated by limited proteolysis, we have studied the structural requirements of the enzyme for regulation by these compounds. The deletion mutant of PLC delta1 lacking the first 58 amino acids and the mutant lacking the entire pleckstrin homology (PH) domain were fully active in the detergent assay, and their activities were affected by spermine, sphingosine, Ca2+ and sphingomyelin to the same extent as the native enzyme. The limited proteolysis of PLC delta1 generated two fragments of 40 kDa and 30 kDa, which formed a stable active complex. The relationship between Ca2+ concentration and enzymatic activity was almost identical for the native PLC delta1 and the proteolytic complex. The activity of the proteolytic complex formed by the 40 kDa and 30 kDa peptides was not affected by spermine and sphingosine. Sphingomyelin inhibited the complex slightly less than the native PLC delta1, and this inhibition was not promoted by spermine. These observations suggest that for activation of PLC delta1 by spermine and sphingosine, the region spanning domains of high conservation, named X and Y, must be intact. In contrast, the PH domain and the intact spanning region of the X and Y domains are not essential for inhibition of PLC delta1 by sphingomyelin. PMID- 9346304 TI - Ornithine carbamoyltransferase from the extreme thermophile Thermus thermophilus- analysis of the gene and characterisation of the protein. AB - The ornithine carbamoyltransferase (OTC) gene from Thermus thermophilus was cloned from a lambda-ZAP genomic library. An ORF of 903 bp was found coding for a protein of Mr 33,200. The coding region has a very high overall G+C content of 68.0%. T. thermophilus OTC displays 38-48% amino acid identity with other OTC, the most closely related proteins being OTC from the archaeon Pyrococcus furiosus and from Bacillus subtilis. The enzyme was expressed in Escherichia coli and purified to homogeneity using a thermoshock followed by affinity chromatography on delta-N-phosphonoacetyl-L-ornithine-Sepharose. The native enzyme has an Mr of about 110,000, suggesting a trimeric structure, as for most anabolic OTC from various organisms. T. thermophilus OTC exhibits Michaelis-Menten kinetics for carbamoyl phosphate and ornithine with a Km(app) of 0.10 mM for both substrates. The pH optimum was dependent on ornithine concentration with an optimum at pH 8 for ornithine concentrations around Km values. Higher concentrations shift the optimum towards lower pH. The optimal temperature was above 65 degrees C and the activation energy 39.1 kJ/mol. The enzyme is highly thermostable. In the presence of its substrates the half-life time was several hours at 85 degrees C. Ionic and hydrophobic interactions contribute to the stability. The expression of T. thermophilus OTC was negatively regulated by arginine. PMID- 9346305 TI - Heparin binding of protein-C inhibitor--analysis of the effect of heparin on the interaction of protein-C inhibitor with tissue kallikrein. AB - The non-specific serine-protease inhibitor protein-C inhibitor (PCI) inactivates its target enzymes by forming stable 1:1 complexes. Heparin stimulates most PCI/protease reactions, but interferes with the inhibition of tissue kallikrein by PCI by a hitherto unknown mechanism. In this study we analyzed the inhibitory effect of heparin on the tissue-kallikrein-PCI interaction. Free PCI and tissue kallikrein x PCI complexes but not free tissue kallikrein bound to heparin Sepharose, implying that the inhibitory effect of heparin cannot be caused by a tissue-kallikrein-heparin interaction. Heparin did not dissociate tissue kallikrein x PCI complexes, making it unlikely that in the presence of heparin PCI becomes a substrate for, rather than an inhibitor of, tissue kallikrein. However, heparin-bound PCI, which was able to form complexes with 125I-urokinase, did not form complexes with 125I-tissue-kallikrein. This suggests that the inhibitory effect of heparin is either based on the neutralization of positive charges in the PCI molecule, which might be required for the interaction of PCI with the acidic protease tissue kallikrein, or on a change in reactivity of PCI upon heparin binding, making heparin-bound PCI no longer a tissue-kallikrein inhibitor. Neutralization of basic amino acids in the PCI molecule by glutamic acid, which prevented in a dose-dependent way the inhibitory effect of heparin, did not have any effect on the tissue-kallikrein-PCI interaction. Therefore, direct involvement of basic amino acid residues present in the heparin-binding site of PCI in the tissue-kallikrein-PCI interaction can be excluded. Heparin binding might rather cause a change in reactivity of PCI (e.g. by inducing a conformational change or by steric interference), thereby preventing its interaction with tissue kallikrein. PMID- 9346306 TI - Identification, isolation and biochemical characterization of a phosphopantetheine:protein transferase that activates the two type-I fatty acid synthases of Brevibacterium ammoniagenes. AB - Upon heterologous expression of the Brevibacterium ammoniagenes type-I fatty acid synthase FAS-A in Escherichia coli, only the pantetheine-free apoenzyme is synthesized. Activation of FAS-A to its holoform was achieved by transformation with a second B. ammoniagenes gene, PPT1, encoding a type-I FAS-specific phosphopantetheine transferase. PPT1 was identified as a coding sequence located immediately downstream of the second FAS gene present on the B. ammoniagenes genome, fasB. Due to this linkage, PPT1 was part of the cloned fasB DNA region and, consequently, FAS-B but not FAS-A was synthesized as holoFAS in E. coli. PPT1 encodes a protein of 153 amino acids and has a calculated molecular mass of 16,884 Da. The PPT1 gene product contains 25% identical and 42% conserved amino acids compared with the type-II acyl-carrier-protein-activating enzyme of E. coli. Although there is essentially no intergenic region between fasB and PPT1, the PPTase gene is autonomously expressed in E. coli if flanked by 200 bp of its endogenous 5' DNA. The structural independence of Ppt1p was confirmed immunologically, as specific antibodies react with the purified PPTase but not with FAS-B. Overexpression and purification of the His-tagged Ppt1p allowed the in vitro activation of apoFAS-A. This holoenzyme synthesis requires, in addition to Ppt1p, CoA and Mg2+ and leads to a specific FAS activity comparable to that of natural B. ammoniagenes FAS-A. The reactivity of the in vitro-activated FAS-A was verified by the optical FAS assay and by analysis of its in vitro products. In agreement with the known overall colinearity of B. ammoniagenes FAS-B and the Saccharomyces cerevisiae FAS1 and FAS2 gene products, a PPT1-like sequence is also observed at the C terminus of FAS2. However, in contrast to B. ammoniagenes PPT1, this sequence is an integral part of the yeast FAS2 gene. Thus, activation of type-I fatty acid synthases may be accomplished by distinct trans-acting PPTase enzymes and by intrinsic cis-acting PPTase domains. PMID- 9346308 TI - Sulfite stimulates the ATP hydrolysis activity of but not proton translocation by the ATP synthase of Rhodobacter capsulatus and interferes with its activation by delta muH+. AB - Sulfite stimulates the rate of ATP hydrolysis by the ATP synthase in chromatophores of Rhodobacter capsulatus. The stimulated activity is inhibited by oligomycin. The activation takes place also in uncoupled chromatophores. The activation consists in an increase of about 12-15-fold of the Vmax for the ATP hydrolysis reaction, while the Km for MgATP is unaffected at 0.16+/-0.03 mM. The dependence of Vmax on the sulfite concentration follows a hyperbolic pattern with half maximum effect at 12 mM. Sulfite affects the ability of the enzyme in translocating protons. Concomitant measurements of the rate of ATP hydrolysis and of ATP-induced protonic flows demonstrate that at sulfite concentrations of greater than 10 mM the hydrolytic reaction becomes progressively uncoupled from the process of proton translocation. This is accompanied by an inhibition of ATP synthesis, either driven by light or by artificially induced ionic gradients. ATP synthesis is totally inhibited at concentrations of at least 80 mM. Sulfite interferes with the mechanism of activation by delta muH+. Low concentrations of this anion (< or = 2 mM) prevent the activation by delta muH+. At higher concentrations a marked stimulation of the activity prevails, regardless of the occurrence of a delta muH+ across the membrane. Phosphate at millimolar concentrations can reverse the inhibition by sulfite. These experimental results can be simulated by a model assuming multiple and competitive equilibria for phosphate or sulfite binding with two binding sites for the two ligands (for sulfite K1S = 0.26 and K2S = 37 mM, and for phosphate K1P = 0.06 and K2P = 4.22 mM), and in which the state bound only to one sulfite molecule is totally inactive in hydrolysis. The competition between phosphate and sulfite is consistent with the molecular structures of the two ligands and of the enzyme. PMID- 9346307 TI - Extensive random mutagenesis analysis of the Na+/K+-ATPase alpha subunit identifies known and previously unidentified amino acid residues that alter ouabain sensitivity--implications for ouabain binding. AB - Random mutagenesis with ouabain selection has been used to comprehensively scan the extracellular and transmembrane domains of the alpha1 subunit of the sheep Na+/K+-ATPase for amino acid residues that alter ouabain sensitivity. The four random mutant libraries used in this study include all of the transmembrane and extracellular regions of the molecule as well as 75% of the cytoplasmic domains. Through an extensive number of HeLa cell transfections of these libraries and subsequent ouabain selection, 24 ouabain-resistant clones have been identified. All previously described amino acids that confer ouabain resistance were identified, confirming the completeness of this random mutagenesis screen. The amino acid substitutions that confer the greatest ouabain resistance, such as Gln111-->Arg, Asp121-->Gly, Asp121-->Glu, Asn122-->Asp, and Thr797-->Ala were identified more than once in this study. This extensive survey of the extracellular and transmembrane regions of the Na+/K+-ATPase molecule has identified two new regions of the molecule that affect ouabain sensitivity: the H4 and the H10 transmembrane regions. The new substitutions identified in this study are Leu330-->Gln, Ala331-->Gly, Thr338-->Ala, and Thr338-->Asn in the H4 transmembrane domain and Phe982-->Ser in the H10 transmembrane domain. These substitutions confer modest increases in the concentration of cardiac glycoside needed to produce 50% inhibition of activity (IC50 values), 3.1-7.9-fold difference. The results of this extensive screening of the Na+/K+-ATPase alpha1 subunit to identify amino acids residues that are important in ouabain sensitivity further supports our hypothesis that the H1-H2 and H4-H8 regions represent the major binding sites for the cardiac glycoside class of drugs. PMID- 9346309 TI - Characterisation of macrophage inflammatory protein-5/human CC cytokine-2, a member of the macrophage-inflammatory-protein family of chemokines. AB - A human monocyte-activating CC chemokine has been identified based on sequences in an expressed sequence tag (EST) cDNA database. The protein shows highest sequence identity to the macrophage inflammatory protein (MIP) group of chemokines, particularly MIP-3 (76.7%) and MIP-1alpha (75.4%), and has been named MIP-5. Model building confirms that the protein has a similar three dimensional structure to other chemokines, but has an additional third disulphide bond. Northern blot analysis and reverse-transcriptase PCR show that the mRNA for MIP-5 is expressed at a high levels in liver, intestine and in lung leukocytes. MIP-5 induces chemotaxis of human monocytes, T-lymphocytes and, to a lesser degree, eosinophils at nanomolar concentrations; it has no effect on neutrophil migration. In receptor-binding assays, MIP-5 shows IC50 values of 12 nM for competition with 125I-MIP-1alpha for binding to CC-chemokine receptor (CCR)1, and 2.5 nM for competition with 125I-MCP-3 for binding to CCR3. It shows no ability to compete with ligand for binding to the two interleukin (IL)-8 receptors (CXC chemokine receptors 1 and 2) or to CCR2, CCR4 or CCR5. Consistent with this binding data, MIP-5 was only able to induce calcium fluxes in CHO cells stably transfected with CCR1 or CCR3. PMID- 9346310 TI - Cloning and expression of the fadH gene and characterization of the gene product 2,4-dienoyl coenzyme A reductase from Escherichia coli. AB - The fadH gene coding for an NADPH-dependent 2.4-dienoyl-CoA reductase from Escherichia coli has been cloned by the polymerase chain reaction. This gene is located at 67.65 min on the E. coli chromosome. The complete open reading frame contains 2019 bp coding for the processed protein of 671 amino acid residues, with a calculated molecular mass of 72.55 kDa, which lacks the N-terminal methionine. Construction and expression of the plasmid pNDH, which contained the fadH gene under the control of the T7 promoter, resulted in a 110-fold increase in the reductase activity above the level detected in E. coli cells containing the control vector. The kinetic parameters of the purified reductase were determined to be 50 microM and 2.3 microM for the Km values of NADPH and 2-trans, 4-trans-decadienoyl-CoA, respectively, and 16 s(-1) for the k(cat) value. Analysis of the kinetic data revealed that the reaction catalyzed by this enzyme proceeds via a ping-pong mechanism. The observed dissimilarity between the E. coli and mammalian 2,4-dienoyl-CoA reductase sequences suggests that they have evolved from distinct ancestral genes. Sequence analysis also suggests that the N terminal part of the E. coli reductase contains the FAD-binding domain whereas the NADPH-binding domain is located in the C-terminal region of the protein. PMID- 9346311 TI - Biosynthesis of the proteoglycan decorin--transient 2-phosphorylation of xylose during formation of the trisaccharide linkage region. AB - Phosphorylation of decorin was investigated by incubating a rat fibroblast cell line with radiolabelled phosphate and carbohydrate precursors. There was a transient phosphorylation of the linkage-region saccharides in intracellular decorin prior to assembly of the galactosaminoglycan chain. Phosphorylation gradually increased from xylosylated, galactosyl-xylosylated to galactosyl galactosyl-xylosylated core protein where all trisaccharide stubs were phosphorylated. Addition of the first glucuronate residue was accompanied by rapid dephosphorylation. Brefeldin A treatment resulted in segregation of galactosaminoglycan synthesis and dephosphorylation. Enzymatic degradation of brefeldin-A-arrested immature proteoglycan with incomplete galactosaminoglycan chain [Moses, J., Oldberg, A., Eklund, E. & Fransson, L.-A. (1997) Eur. J. Biochem., in the press] by using chondroitin AC lyase and chondro-glycuronidase, followed by beta-galactosidase treatment, demonstrated the sequence galactosyl galactosyl-phosphoxylose. The xylose was resistant to direct periodate oxidation, but sensitive after treatment with alkaline phosphatase, showing that the phosphate was located at C2 of xylose. The transient 2-phosphorylation of xylose may be involved in intracellular transport and/or in the control of modifications of the glycan chain. PMID- 9346312 TI - Partial characterization and enrichment of a membrane-bound sialidase specific for gangliosides from human brain tissue. AB - Gangliosides, constituents of surfaces of vertebrate cells, modulate important cellular functions. Ganglioside-specific sialidases that possibly control these processes have been observed in a number of tissues, but their characterization has proved difficult due to their low abundance and lability. Here we describe the partial isolation and characterization of a ganglioside sialidase from human brain grey matter. After membrane extraction with octylglucoside, the enzyme was purified about 1300-fold by ion-exchange, affinity and gel-permeation chromatographies. Although PAGE still showed several protein bands, specific photoaffinity labelling with iodinated 5-N-acetyl-9-(4-azidosalicoylamido)-2,9 dideoxy-2,3-didehydrone uraminic acid identified a single polypeptide of 60 kDa likely to contain the active site of the sialidase. In the presence of 0.4% octylglucoside, the purified sialidase desialylated gangliosides G(M3), G(D1a), G(D1b) and G(T1b), but was inactive towards G(M1), G(M2), colominic acid, sialyl (alpha2-3)-lactose, 2-(4-methylumbelliferyl)-neuraminate, or the glycoprotein fetuin. The ganglioside sialidase activity was strongly inhibited by 2-deoxy-2,3 didehydro-N-acetylneuraminic acid, heparin and heparan sulfate. Because of its substrate and inhibitor profiles, the purified enzyme resembles the activity characterized previously in the plasma membrane of human neuroblastoma cells, but is distinct from a lysosomal activity. The purified brain sialidase thus appears to function in the selective desialylation of gangliosides with terminal sialic acid residues. PMID- 9346313 TI - Molecular cloning and functional expression of an insect high-affinity Na+ dependent glutamate transporter. AB - Excitatory amino acid transporters in the central and peripheral nervous systems of insects are thought to assist in maintaining glutamate concentrations in the resting synapse below the activation threshold of glutamate receptors. We have isolated a cDNA from the caterpillar Trichoplusia ni which encodes a high affinity Na+-dependent glutamate transporter, designated TrnEAAT1. The deduced amino acid sequence shows strong identity with known members of the vertebrate Na+- and K+-dependent amino acid transporter family. Expression of the insect transporter mRNA was predominantly localized in the caterpillar brain. The function of the TrnEAAT1 protein was analyzed in cultured insect cells using a baculovirus expression system. Cells infected with the recombinant virus were found to exhibit a 50-fold increase in ability to accumulate labeled L-glutamate compared to mock-infected cultures, and this activity was shown to be Na+ dependent. Transport activity was further demonstrated by chromatographic identification of various glutamate analogues accumulated by infected cells. Various glutamate uptake inhibitors were used to outline the pharmacological properties of the cloned transporter and to compare it with known mammalian transporters. Despite the significant differences between insect and vertebrate physiology, the characteristics of the respective transporters were found to be remarkably similar. PMID- 9346314 TI - Sequence-specific antiproliferative effects of antisense and end-capping-modified antisense oligodeoxynucleotides targeted against the 5'-terminus of basic fibroblast-growth-factor mRNA in coronary smooth muscle cells. AB - Basic fibroblast growth factor (bFGF), a potent mitogen for arterial smooth muscle cells, has been shown to play a fundamental role in the pathogenesis of arteriosclerosis and restenosis by stimulating the proliferation of vascular smooth muscle cells. We found that partially phosphorothioate-modified 15-residue antisense oligodeoxynucleotides complementary to bFGF mRNA at 0.1-2.0 microM block growth and division of cultured human and bovine coronary smooth muscle cells in a dose-dependent manner. The effect is sequence specific at low (0.1-0.5 microM) nontoxic concentrations. It is associated with inhibition of expression of pericellular and intracellular bFGF, with a decreased de novo synthesis of bFGF and is partly reversible by the addition of exogenous (recombinant) bFGF. The antisense effect lasts 48-72 h and diminishes thereafter. If the antisense oligodeoxynucleotide medium is replaced by an oligonucleotide-free medium after 24 h, the [3H]thymidine incorporation rate returns to control levels. Under the same conditions, the corresponding sense oligodeoxynucleotide exerts negligible nonspecific inhibitory actions. The antiproliferative potency of the 15-residue antisense oligodeoxynucleotide is markedly enhanced by adding 3-4 nonbase-pairing guanosine residues at the 5'- and 3'-termini of the 15-residue antisense oligonucleotide. The data implicate bFGF in the process of smooth muscle cell proliferation and an effective and specific antiproliferative potency of bFGF specific antisense oligonucleotides. The results point to possible new therapeutic strategies for the use of antisense methodology in the suppression of post-angioplasty restenosis. PMID- 9346315 TI - Molecular cloning and expression of bothrojaracin, a potent thrombin inhibitor from snake venom. AB - Bothrojaracin is a potent and selective thrombin inhibitor that has been isolated from the venom of Bothrops jararaca. It does not interact with the catalytic site of the enzyme but binds to both anion-binding exosites 1 and 2 resulting in a potent inhibition of thrombin activity towards fibrinogen and platelets [Zingali, R. B., Jandrot-Perrus, M., Guillin, M. C. & Bon, C. (1993) Biochemistry 32, 10794 108021. Bothrojaracin is a 27-kDa protein composed of two disulfide-linked polypeptide chains, A and B, of 15 kDa and 13 kDa, respectively. The sequences of A and B chains determined by molecular cloning exhibit a high degree of identity with other snake venom lectin-like proteins. In contrast to other ligands that interact with thrombin exosite 1, the amino acid sequence of bothrojaracin does not contain an acidic sequence similar to the C-terminal tail of hirudin. Expression of functional bothrojaracin was achieved in COS cells upon transfection with two pcDNA3 vectors containing the complete cDNAs. Recombinant bothrojaracin, which was secreted into the medium, was able to bind to and inhibit thrombin. When expressed alone, the B chain formed inactive dimers that were secreted into the culture medium. In contrast, no bothrojaracin-related protein was detected in conditioned media from cells transfected with the A chain. PMID- 9346316 TI - 69-kDa and 100-kDa isoforms of interferon-induced (2'-5')oligoadenylate synthetase exhibit differential catalytic parameters. AB - The (2'-5')oligoadenylate synthetase represents a family of interferon-induced proteins which when activated by double-stranded (ds)RNA polymerizes ATP into 2' 5'-linked oligomers with the general formula pppA(2'p5'A)n, where n > 1, which for convenience are referred to as 2-5A. We studied here the influence of pH, dsRNA concentration and time on oligomeric composition of 2-5A synthesized by purified 69-kDa and 100-kDa isoforms of (2'-5')oligo(adenylate) synthetase. In optimal conditions for activity, the 69-kDa form synthesized higher oligomers of 2-5A molecules whereas the 100 kDa form synthesized preferentially dimeric molecules, which are known not to be functional for the activation of RNase L. This difference does not reflect a differential affinity of the enzymes for the preformed 2-5A dimer, which is found to be a very poor substrate for both enzymes. This latter strongly suggests that the mechanism of elongation is more likely processive. Moreover, we show that both isoforms have efficient nucleotidyl-transferase activity and provide evidence that, in optimized conditions, GTP can be used alone as substrate by these enzymes to generate pppG2'p5'G. Our results clearly demonstrate that the 69-kDa and 100-kDa forms of (2'-5')oligoadenylate synthetase manifest various differential catalytic activities, and favor the hypothesis that these enzymes might have other functions in the cell besides those in the 2-5A system. PMID- 9346317 TI - The effect of high pressure on thermolysin. AB - The effects of high pressure on thermolysin activity and spectroscopic properties were studied. Thermolysin showed distinct pressure-induced activation with a maximum observed at 200-250 MPa for a dipeptide amide substrate and at 100-120 MPa for a heptapeptide substrate. By examining the pressure dependence of the hydrolytic rate for the former substrate using a high pressure stopped-flow apparatus as a mixing device under elevated pressures, the activation volume of the reaction was -71 ml mol(-1) at 25 degrees C. Delta V++ was accompanied by a negative activation expansibility and a value of -95 ml mol(-1) was obtained at 45 degrees C. A prolonged incubation of thermolysin under high pressure, however, caused a time-dependent deactivation. These changes due to pressure were monitored by several spectroscopic methods. The fourth-derivative absorbance spectrum showed an irreversible change, mostly in the tyrosine and tryptophan regions, at a pressure higher than 300 MPa. Intrinsic fluorescence and circular dichroism measurements of thermolysin in solution also detected irreversible changes. All these measurements indicated that a change occurred at higher pressures and are explained by a simple two-state transition model accompanied by a large, negative change in the volume of reaction. PMID- 9346318 TI - Identification of a region required for binding to presecretory protein in Bacillus subtilis Ffh, a homologue of the 54-kDa subunit of mammalian signal recognition particle. AB - Bacillus subtilis Ffh protein is a homologue of the 54-kDa subunit of mammalian signal recognition particle (SRP54). It contains three highly hydrophobic regions (h1, h2, and h3) in the C-terminal methionine-rich domain (M-domain). Two of the hydrophobic regions, h2 and h3, are essential for small cytoplasmic RNA (scRNA) binding [Kurita, K., Honda, K., Suzuma, S., Takamatsu, H., Nakamura, K., & Yamane, K. (1996) J. Biol. Chem. 271, 13,140-13,146]. Using purified presecretory proteins and mutant Ffh proteins, we identified a region required for presecretory protein binding in B. subtilis Ffh. Deletion of this region, which consisted of residues Ser311-Gly362 of B. subtilis Ffh, including a hydrophobic sequence (h1), reduced precursor binding activity. In contrast, deletions of residues Leu121-Lys279, Lys364-Met446, or Leu338-Ser397 of B. subtilis Ffh did not. We also analyzed the mutant B. subtilis Ffh proteins, FfhQQQR and FfhQQQQ having wild-type residues 398-401 (Arg-Arg-Lys-Arg) replaced with Gln3Arg and Gln4, respectively. FfhQQQR bound to both scRNA and presecretory protein. Although the FfhQQQQ mutation prevented binding to scRNA, binding to the precursor was not affected. FfhQQQR restored the growth of B. subtilis DF46 strain in which ffh gene expression is regulated by an inducible promoter in the absence of an inducer, whereas FfhQQQQ did not. These results indicate that the region including h1 is required for B. subtilis Ffh to bind to presecretory protein. The results also suggest that scRNA is required for the complete function of the B. subtilis SRP-like particle in vivo, although this protein is intrinsically capable of binding a signal peptide free from scRNA. PMID- 9346319 TI - Breast carcinoma epithelial cells express a very low-density lipoprotein receptor variant lacking the O-linked glycosylation domain encoded by exon 16, but with full binding activity for serine proteinase/serpin complexes and Mr-40,000 receptor-associated protein. AB - Very-low density lipoprotein receptor (VLDLR) belongs to the low-density lipoprotein receptor family of endocytosis receptors. It binds a variety of different ligands, including apolipoprotein E, Mr-40,000 receptor-associated protein (RAP), and some serine proteinase/serpin complexes. We previously demonstrated the occurrence of two forms of VLDLR in SDS/PAGE, migrating with Mr 105,000 and Mr 130,000, respectively [Heegaard, C. W., Simonsen, A. C. W., Oka, K., Kjoller, L., Christensen, A., Madsen, B., Ellgaard, L., Chan, L. & Andreasen, P. A. (1995) J. Biol. Chem. 270, 20,855-20,869]. We now demonstrate that these two forms correspond to forms with the absence (type-II) and presence (type-I) of the O-linked glycosylation domain encoded by exon 16, respectively. We show that the two forms have the same binding affinity to RAP and serine proteinase/serpin complexes. Using reverse transcription and PCR, we demonstrate that the splice variation giving rise to the two forms is highly cell specific. In particular, we demonstrate that human breast carcinomas express predominantly or exclusively the variant lacking exon 16. By immunohistochemistry, we demonstrate that VLDLR is mainly expressed by the epithelial cancer cells in these carcinomas. The VLDLR variant expressed by epithelial cancer cells could function in the clearance of cell-surface-associated serine proteinase/serpin complexes in breast carcinomas. PMID- 9346320 TI - Lipopolysaccharides of Helicobacter pylori serogroups O:3 and O:6--structures of a class of lipopolysaccharides with reference to the location of oligomeric units of D-glycero-alpha-D-manno-heptose residues. AB - Lipopolysaccharides (LPS) from antigenically different strains assigned to serogroups O:3 and O:6 of Helicobacter pylori were isolated as water-soluble material of high Mr and as water-insoluble gels of low Mr. Chemical and spectroscopic analyses of the soluble LPS and oligosaccharides liberated from the water-insoluble gels led to proposed structures with Lewis (Le) antigen determinants terminating regular repeating units of different types, linked in turn to inner core regions of invariable structure. The O:6 LPS has two populations of related molecules with chains of 3-linked D-glycero-alpha-D-manno heptose residues similar to those in the MO19 strain, one with and the other without a single terminal Lewis (Le(y)) epitope. In contrast, in the O:3 LPS, Lewis (Le(x) and Le(y)) epitopes terminate a partially fucosylated N acetyllactosaminoglycan, but a heptan chain similar to that in the O:6 LPS was shown to connect the outer chains to the inner core. These LPS provide examples of the molecular mimicry of cell-surface glycoconjugates. Structural variations of LPS between strains, and differences in some aspects of structure within strains, between high Mr and low Mr LPS indicate a class of LPS whose mechanisms of biosynthesis lead to overall architectures different from those characteristic of most LPS from enteric bacteria. PMID- 9346321 TI - Mass determination, subunit organization and control of oligomerization states of keyhole limpet hemocyanin (KLH). AB - Analytical dark-field scanning transmission electron microscopy (STEM) of freeze dried unstained specimens of keyhole limpet hemocyanin (KLH; from Megathura crenulata, a prosobranch gastropod) gave a molecular mass of 400 kDa for the subunit of KLH1 and of 345 kDa for the subunit of KLH2, which confirms our published values from SDS/PAGE. Within the 400-kDa KLH1 subunit we identified, by limited proteolysis, isolation of fragments and N-terminal sequencing, eight distinct 45-60 kDa functional domains (termed 1a through 1h) and determined their sequential arrangement. The KLH1 domains differ biochemically and immunologically from each other and from the previously characterized seven domains of KLH2 (termed 2a through 2g). Our partial amino acid sequences suggest that a domain, equivalent to the C-terminal domain 1h, is missing in KLH2. This deficiency is believed to be genuine and not an artifact of the subunit preparation procedure, since STEM measurements of the native didecamers yielded a mass difference of about 800 kDa between KLH1 and KLH2 (8.3 MDa versus 7.5 MDa), correlating with 20 copies of a functional 1h domain. It was also shown that the KLH1 didecamer can be rapidly split (minutes) into an almost homogeneous population of stable decamers by increasing the pH of the Tris/saline stabilizing buffer (routinely pH 7.4), which contains 5 mM CaCl2 and 5 mM MgCl2, to pH 8.5. Reformation of the didecamers occurred more slowly (days) upon dialysis against the pH 7.4 stabilizing buffer. Addition of 100 mM calcium and 100 mM magnesium ions to the pH 7.4 stabilizing buffer leads to the more rapid (overnight) formation of didecamers together with a significant number of previously unobserved KLH1 multidecamers, which could be structurally distinguished from the established multidecamers of KLH2. PMID- 9346322 TI - Functional characteristics of heterologously expressed 5-HT receptors. AB - Over the past 10 years, molecular cloning has revealed the presence of 15 serotonin (5-hydroxytryptamine; 5-HT) receptor subtypes, which can be subdivided in seven subfamilies. Except for the 5-HT3 receptors, which are ligand-gated ion channels, all 5-HT receptors belong to the superfamily of G-protein-coupled receptors. The large multiplicity of 5-HT receptor subtypes has been suggested to be a direct result of the evolutionary age of the 5-HT system. Molecular information on G-protein-coupled 5-HT receptors is currently available for several mammalian species as well as for a limited number of invertebrate species (insects, molluscs). The aim of this review is to give an overview of all cloned 5-HT receptor subtypes belonging to the superfamily of G-protein-coupled receptors with specific emphasis on the pharmacological and signaling properties of the receptors upon expression in several heterologous expression systems. PMID- 9346323 TI - Repeated DOI and SR 46349B treatments do not affect elevated plus-maze anxiety despite opposite effects on cortical 5-HT2A receptors. AB - We report the consequences of a 4-day treatment (b.i.d) with the 5-HT2A,2B,2C receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI, 1.5 mg/kg) or the selective 5-HT2A receptor antagonist trans-4-[(3Z)3-(2 dimethylaminoethyl)oxyimino-3-(2-fluorophe nyl)propen-1-yl]phenol hemifumarate (SR 46349B, 7.5 mg/kg) on (i) anxiety-related behaviour in an elevated plus-maze, and (ii) specific [3H]ketanserin binding at central 5-HT2A receptors, in Roman rats. Neither DOI nor SR 46349B pretreatment affected the behaviour in the open arms of the elevated plus-maze; however, DOI pretreatment promoted discrete changes in the closed arm entries. The Bmax value of [3H]ketanserin binding at cortical 5-HT2A receptors was decreased by repeated DOI pretreatment. Conversely, Bmax, but also KD, values were increased by SR 46349B pretreatment. Thus, changes at central 5-HT2A receptors may occur without there being changes in anxiety related behaviour in the elevated plus-maze. PMID- 9346324 TI - Effects of monoamine oxidase and catechol-O-methyltransferase inhibition on dopamine turnover: a PET study with 6-[18F]L-DOPA. AB - The consequences of monoamine oxidase and catechol-O-methyltransferase inhibition on the effective turnover of dopamine were investigated using 6-[18F]L-3-4 dihydroxyphenylalanine (6-[18F]L-DOPA) and positron emission tomography. The effective dopamine turnover was expressed as the ratio between the rate of reversibility of 6-[18F]L-DOPA trapping (k[loss]) and the rate of uptake of 6 [81F]L-DOPA (Ki) in the striatum of normal cynomolgus monkeys. The monkeys received 6-[18F]L-DOPA scans, untreated or after pretreatment with either the peripheral catechol-O-methyltransferase inhibitor nitecapone; the peripheral and central catechol-O-methyltransferase inhibitor tolcapone; the monoamine oxidase inhibitors deprenyl or pargyline; a combination of tolcapone and the monoamine oxidase inhibitors. Tolcapone alone or combined with the monoamine oxidase inhibitors produced a significant decrease in the dopamine turnover (55 to 65%). Neither nitecapone nor monoamine oxidase inhibition alone produced significant changes. These results may have implications for the use of central catechol-O methyltransferase inhibitors added to routine levodopa therapy in parkinsonian patients. PMID- 9346325 TI - Sub-anesthetic doses of bupivacaine or lidocaine increase peripheral ICS-205 930 induced analgesia against inflammatory pain in rats. AB - Intraplantar co-administration of sub-anesthetic doses of bupivacaine (2.5 microg) or lidocaine (7.5 microg) increased the dose- and time-dependent analgesic effects of the 5-HT3 receptor antagonist, 3-a-tropanyl-1-H-indole-3 carboxylic ester (ICS-205 930) (1-100 microg; 50 microl) against inflammatory pain induced by hindpaw inoculation with complete Freund's adjuvant. The effects of bupivacaine were greater than lidocaine at all doses of ICS-205 930 tested. These findings may reflect facilitation of ICS-205 930 effects through negative allosteric modulation by bupivacaine and lidocaine of peripheral 5-HT3 receptors involved in nociceptive processing. PMID- 9346326 TI - The muscarinic toxin 3 augments neuropeptide mRNA in rat striatum in vivo. AB - The selective M4 muscarinic receptor toxin, MT3, was used in vivo to evaluate the role of M4 receptors in cholinergic inhibition of neuropeptide mRNA expression in striatonigral neurons. Unilateral injection of the muscarinic toxin 3 (0.04-4 nmol) into the dorsal striatum of chronically-cannulated rats elevated basal levels of preprodynorphin, substance P and preproenkephalin mRNAs in the ipsilateral dorsal striatum as revealed by quantitative in situ hybridization. Pretreatment with muscarinic toxin 3 also augmented amphetamine (2.5 mg/kg, i.p.) stimulated preprodynorphin and substance P expression in the dorsal striatum in a manner similar to that observed after the muscarinic antagonist, scopolamine. Since muscarinic toxin 3 has a much greater affinity for muscarinic M4 receptors than for other subtypes, it is possible that muscarinic toxin 3, by interacting with the muscarinic M4 subtype, regulates basal and/or dopamine-stimulated striatal neuropeptide gene expression. PMID- 9346328 TI - Endothelin contributes to the hemodynamic effects of vasopressin in spontaneous hypertension. AB - Changes in blood pressure, cardiac output, and total peripheral conductance evoked by intravenous infusions of [Arg8]-vasopressin (vasopressin) were recorded before and after pretreatment with bosentan, a non-selective endothelin antagonist, in conscious unrestrained spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). The presser effects of vasopressin were exaggerated in SHR compared to WKY. Pretreatment with bosentan failed to change hemodynamic responses of WKY to vasopressin, but it blunted the increases in blood pressure and the decreases in conductance evoked by vasopressin in SHR. In contrast, bosentan failed to change cardiac output responses of SHR to vasopressin. Except at the highest dose of vasopressin, bosentan abolished the exaggerated pressor responsiveness of the SHR to vasopressin. The results suggest that endothelin contributes to the exaggerated pressor responsiveness of SHR to vasopressin, and that this effect is exerted at the level of the resistance vessels and not on factors that regulate cardiac output. PMID- 9346327 TI - Contribution of endopeptidase 3.4.24.15 to central neurotensin inactivation. AB - The tridecapeptide, neurotensin elicits naloxone-insensitive analgesia after its intracebroventricular administration in mice. We used this central pharmacological effect to assess the putative contribution of the endopeptidase 3.4.24.15 to central inactivation of the peptide. By means of combinatorial chemistry, we previously designed the first potent endopeptidase 3.4.24.15 inhibitor. This agent, Z-(L,D)Phe psi(PO2CH2)(L,D)Ala-Lys-Met (phosphodiepryl 21), is shown here to behave as a fully specific endopeptidase 3.4.24.15 inhibitor, as demonstrated by the absence of effect on a series of other exo- and endopeptidases belonging to various classes of proteolytic activities present in murine brain membranes. Furthermore, central administration of phosphodiepryl 21 drastically prolongs the forepaw licking latency of mice tested on the hot plate and injected with sub-maximally active doses of neurotensin. Altogether, our results demonstrated that, in addition to endopeptidase 3.4.24.16, endopeptidase 3.4.24.15 likely contributes to the physiological termination of the neurotensinergic message in murine brain. PMID- 9346329 TI - Distinct receptors mediate pituitary adenylate cyclase-activating peptide- and vasoactive intestinal peptide-induced relaxation of rat ileal longitudinal muscle. AB - Relaxant responses to pituitary adenylate cyclase-activating peptide (PACAP)-27, PACAP-38 and vasoactive intestinal peptide (VIP) were examined in rat ileal longitudinal muscle. PACAP-27 was much more potent than PACAP-38 and VIP, with PACAP-38 and VIP being equipotent. The relaxation induced by each of the peptides was unaffected by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) (10[-4] M), tetrodotoxin (10[-6] M) or atropine (10[-6] M). Pretreatment with apamin (10[-6] M) abolished the relaxations induced by PACAP-27, but not those induced by PACAP-38 or VIP. Pretreatment with neuropeptide Y (NPY) (10[-7] M) inhibited relaxations induced by VIP, but not those induced by PACAP-27 or PACAP 38. No cross-desensitization between PACAP-27 and VIP could be revealed. In conclusion, distinct receptors mediate PACAP- and VIP-induced relaxations of rat ileal longitudinal muscle. At least three different types of receptors may exist: (1) a PACAP-27 preferring receptor coupled to apamin sensitive Ca2+-dependent K+ channels, (2) a PACAP specific receptor activated by both PACAP-27 and PACAP-38 but not by VIP and (3) a VIP specific receptor regulated by NPY by yet unknown mechanisms. PMID- 9346330 TI - Effects of neuropeptide FF on intestinal motility and temperature changes induced by endotoxin and platelet-activating factor. AB - Several effects of bacterial endotoxins involve an opioid pathway and neuropeptide FF is an endogenous peptide known to modulate opioid activity, mainly in the central nervous system. The aim of this study was to investigate in rats the role of central neuropeptide FF receptors in intestinal motor disturbances and body temperature changes induced by endotoxins and platelet activating factor (PAF), a major endotoxin mediator. Rats were fitted with intestinal electrodes, an intraperitoneal thermistor probe and an intracerebroventricular (i.c.v.) cannula for long-term use. E. coli endotoxin (100 microg/kg, i.v.) disrupted the cyclic pattern of intestinal migrating myoelectric complexes and induced a biphasic increase in body temperature while PAF (25 microg/kg, i.p.) disrupted the migrating myoelectric complexes and induced hypothermia for about 2 h. The neuropeptide FF analog, (1 DME)Y8Fa (D-Tyr D-Leu[N-Me]-Phe-Gln-Pro-Gln-Arg-Phe-NH2) administered i.c.v. 40 and 100 microg/kg reduced the duration of migrating myoelectric complex disruption induced by endotoxin and PAF and abolished the PAF-induced hypothermia. Only at the dose of 100 microg/kg did (1 DME)Y8Fa change the biphasic endotoxin-induced hyperthermia into a monophasic increase. Naloxone (1 mg/kg, s.c.) reduced only the duration of migrating myoelectric complex disruption induced by endotoxin. These results indicate that central neuropeptide FF modulates the intestinal motor disturbances and changes in body temperature induced by endotoxin and PAF. Its action against endotoxin may involve an anti-opioid pathway whereas its action against PAF does not. PMID- 9346331 TI - Effects of SCA40 on bovine trachealis muscle and on cyclic nucleotide phosphodiesterases. AB - While UK-93,928 (1-[[3-(6,9-dihydro-6-oxo-9-propyl-1H-purin-2-yl)-4-ethoxyphenyl] sulfonyl]-4-methylpiperazine; 5 nM-5 microM) was devoid of relaxant activity, benzafentrine, isoprenaline, levcromakalim and SCA40 (6-bromo-8 methylaminoimidazo[1,2-a]pyrazine-2-carbonitrile) each relaxed histamine (460 microM)-precontracted bovine isolated trachealis. Each of these relaxants was antagonised by a K+-rich (80 mM) medium. Except in the case of levcromakalim, nifedipine (1 microM) offset this antagonism. Charybdotoxin (100 nM) antagonised isoprenaline in a nifedipine-sensitive manner but did not antagonise SCA40 or benzafentrine. Iberiotoxin (100 nM) did not antagonise SCA40. Acting on tissue precontracted with carbachol, SCA40 potentiated isoprenaline but did not potentiate sodium nitroprusside. While levcromakalim (1 and 10 microM) induced hyperpolarisation, SCA40 (1 and 10 microM) induced little change in the membrane potential of bovine trachealis. In trachealis preloaded with 86Rb+, levcromakalim (1 and 10 microM) promoted efflux of the radiotracer while SCA40 (1 and 10 microM) had no effect. Tested as an inhibitor of isoenzymes of cyclic nucleotide phosphodiesterase, SCA40 was most potent against the type III, less potent against the type IV and least potent against the type I isoenzyme. It is concluded that neither inhibition of phosphodiesterase type V nor the promotion of BKCa channel opening explains the tracheal smooth muscle relaxant activity of SCA40. This compound relaxes bovine tracheal smooth muscle mainly by inhibiting phosphodiesterase isoenzyme types III and IV. PMID- 9346332 TI - Evidence against nitrergic neuromodulation in the rat vas deferens. AB - Electrical field stimulation (60 V, 1 ms, single pulses or 20 s trains of 1-10 Hz) of the nerve terminals within the rat vas deferens produced biphasic contractions in preparations oriented to measure either longitudinal or circular muscle contractions. In confirmation of earlier reports, these contractions were blocked by tetrodotoxin (1 microM). The initial fast purinergic contraction was dominant in prostatic halves of the vas deferens while the second slower noradrenergic contraction was greater in epididymal halves. Although previous studies have shown nitric oxide synthase immuno-positive nerves in the vas deferens, electrical field stimulation-induced contractions were unaffected by L arginine, sodium nitroprusside, N-nitro-L-arginine methyl ester (L-NAME) or superoxide dismutase in concentrations up to I mM. In concentrations above 1 mM, L-NAME reduced the size of the field stimulation-induced contractions but this effect could not be reversed by either L-arginine or sodium nitroprusside. Furthermore, L-arginine, sodium nitroprusside and L-NAME did not affect the contractions induced by exogenous application of noradrenaline (10 microM), ATP (1 mM) or BaCl2 (1-10 mM). We conclude that nitric oxide does not act as a neuromodulator in isolated preparations of rat vas deferens. PMID- 9346333 TI - Relaxation of the ovine isolated iris sphincter by adenosine receptor agonists: lack of effect of adenosine A1 and A2 receptor antagonists. AB - The effects of adenosine receptor ligands on the tone of the ovine isolated iris sphincter were investigated, and adenosine analogues were found to relax the carbachol-contracted tissue in a concentration-dependent manner with an order of potency of 5'-N-ethylcarboxamidoadenosine > or = 2-(p-(2 carboxyethyl)phenylethylamino)-5'-N-ethylcarboxamidoadenos ine (CGS 21680) > or = N6-cyclopentyladenosine > adenosine, consistent with activation of an adenosine A2A receptor. However, these responses were not inhibited by the non-selective adenosine A1/A2 receptor antagonist 8-p-sulphophenyltheophylline (50 microM), the selective adenosine A2A/A1 receptor antagonist N-[2-(dimethylamino)ethyl]-N methyl-4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3- dipropyl-1H-purin-8 yl)benzenesulphonamide (PD 115,199) (0.1 microM) or the non-xanthine adenosine A2A receptor antagonist (4-(2-[7-amino-2-(2-furyl) [1,2,4]-triazolo[2,3 a][1,3,5]triazin-5-yl-amino]ethyl)phenol) (ZM 241385) (0.1 microM). The relaxations cannot therefore be mediated by activation of adenosine A1, A2A or A2B receptors. PMID- 9346334 TI - Actions of isoform-selective and non-selective nitric oxide synthase inhibitors on endotoxin-induced vascular leakage in rat colon. AB - The effects of the nitric oxide (NO) synthase inhibitor, N-(3 (aminomethyl)benzyl)-acetamidine (1400W) which is selective for the inducible isoform of NO synthase, on rat colonic microvascular injury provoked by Escherichia coli endotoxin (3 mg/kg i.v.) has been compared to those of aminoguanidine (25-50 mg/kg, s.c.), NG-iminoethyl-L-ornithine (L-NIO, 15-30 mg/kg, s.c.) and NG-nitro-L-arginine methyl ester (L-NAME, 2-5 mg/kg, s.c.). Administration of aminoguanidine, L-NIO or L-NAME concurrently with endotoxin provoked microvascular albumin leakage 1 h later, presumably by inhibiting constitutive NO synthase, whereas 1400W (0.1-10 mg/kg, s.c.) had no such effect. Administration of all these agents during the expression of inducible NO synthase (i.e. 3 h after endotoxin challenge) attenuated the subsequent endotoxin-provoked albumin leakage 1 h later. Moreover, concurrent administration of 1400W (0.2-5 mg/kg, s.c.; doses that did not affect systemic arterial blood pressure) with endotoxin suppressed the subsequent rise in albumin leakage after 5 h. These findings indicate that 1400W is a potent inhibitor of colonic microvascular injury associated with induction of NO synthase in vivo. 1400W will thus be useful to investigate in vivo the therapeutic potential of a selective inducible NO synthase inhibitor in inflammation. PMID- 9346335 TI - Effect of selected triterpenoids on chronic dermal inflammation. AB - The activity of four natural triterpenoids on a 12-O-tetradecanoylphorbol-13 acetate multiple-dose model of skin chronic inflammation was studied. Erythrodiol and ursolic acid were significantly effective. The most important features concerning structure-activity relationship and previous data on the effect of these triterpenoids on other inflammatory conditions are discussed. PMID- 9346336 TI - Enhancement of maximal activation of neuronal nitric oxide synthase at muscarinic M1 receptors following prolonged agonist treatment. AB - It was previously believed that the neuronal type of nitric oxide (NO) synthase was constitutive in nature, and that changes in the concentration of intracellular Ca2+ represent the sole input that regulates its activity. Recent reports, however, suggested that this enzyme could also be induced under certain conditions. We report here that prolonged stimulation of M1 muscarinic acetylcholine receptors results in potentiation of maximal receptor-mediated activation of neuronal NO synthase in Chinese hamster ovary cells. This effect was dependent on the concentration of agonist during the treatment and was abolished by a muscarinic receptor antagonist. These findings are important for understanding the sequelae of prolonged administration of muscarinic agonists in vivo. PMID- 9346337 TI - Oxygen radicals diminish dopamine transporter function in rat striatum. AB - Incubation of striatal synaptosomes with the oxygen radical generating enzyme, xanthine oxidase, decreased [3H]dopamine uptake: an effect attributable to a decreased Vmax. Concurrent incubation with the superoxide radical scavenger, superoxide dismutase, abolished the xanthine oxidase-induced decrease. These results indicate that, like methamphetamine administration in vivo, reactive oxygen species diminish dopamine transporter function in vitro. The significance of these findings to mechanisms responsible for effects of methamphetamine is discussed. PMID- 9346338 TI - Probing human beta1- and beta2 -adrenoceptors with domain-specific fusion protein antibodies. AB - In order to generate antibodies suitable for immunological studies on beta adrenoceptors constitutively expressed at low levels in cells or tissues we have produced fusion proteins of the amino- and carboxy-terminus, and the second extracellular loop of the human beta1- or beta2-adrenoceptors with bacterial glutathione-S-transferase in E. coli. Rabbit antibodies raised against these fusion proteins strongly reacted with intact human beta1- or beta2-adrenoceptors in a subtype- and domain-specific manner. Antibodies directed against the second extracellular loop of the beta1-adrenoceptor reacted stronger with non-denatured receptors and decreased the affinity of the 3H-labelled antagonist (-)-4-(3-t butylamino-2-hydroxypropoxy)-[5,7-3H]benzimidazol-2-one ([3H]CGP 12 177), indicating a specific interaction with the native receptor. In contrast, antibodies directed against carboxy- and amino-terminal receptor domains reacted strongly both with denatured and non-denatured receptors but did not interfere with binding of [3H]CGP 12 177. Affinity purified antibodies were used for detecting the beta1- or the beta2-adrenoceptor subtype heterologously produced in Sf9 cells by enzyme-linked immunosorbent assay, Western blotting, immunoprecipitation, and indirect immunofluorescence microscopy. Moreover, we could demonstrate that avidity, titers, and specificity of these antibodies were high enough for studying beta-adrenoceptors constitutively expressed in human A431 cells, where we observed a patched membrane distribution of the receptors. PMID- 9346339 TI - SR141716A is an inverse agonist at the human cannabinoid CB1 receptor. AB - The effects of R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3 de]-1,4- benzoxazin-yl]-(1-napthalenyl)methanone mesylate (WIN 55,212-2) and N piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3-pyrazo le carboxamide (SR141716A) on guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding to membranes isolated from human cannabinoid CB1 receptor-transfected Chinese hamster ovary (CHO) cells were examined. WIN 55,212 2 stimulated [35S]GTPgammaS binding 76.3% above basal levels whereas SR141716A produced a 22.3% decrease in basal [35S]GTPgammaS binding. These findings demonstrate that WIN 55,212-2 is an agonist and SR141716A is an inverse agonist in this system. PMID- 9346340 TI - Novel persistent activation of muscarinic M1 receptors by xanomeline. AB - The muscarinic agonists, xanomeline and carbachol, displayed similar intrinsic activities in stimulating neuronal nitric oxide synthase at muscarinic M1 receptors in Chinese hamster ovary (CHO) cells, with xanomeline being more potent. Pre-incubation (1 h) with 1 microM xanomeline, followed by extensive washing, resulted in a significantly elevated basal response, which was absent on co-incubation with atropine. This phenomenon was not observed with carbachol. This is the first report of a persistent, receptor-activating effect of a muscarinic agonist. PMID- 9346341 TI - Status of patent foramen ovale, atrial septal aneurysm, atrial septal defect and aortic arch atheroma as risk factors for stroke. AB - During the last few years, there has been emphasis on the role of the interatrial septum and the aortic arch atheroma in the genesis of stroke. The causal relationship of these disorders with stroke is still controversial because direct evidence may be lacking. The diagnosis is often presumed and based on indirect signs, i.e. by visualization of the interatrial septal or aortic arch abnormalities which are detectable by transesophageal echocardiography. The optimal therapeutic strategy including some more invasive treatments, such as surgery, is uncertain. In this article, we review the current knowledge concerning the interatrial septum and aortic arch disorders as risk factors for stroke and current therapeutic strategies. PMID- 9346342 TI - The Minorities Risk Factors and Stroke Study (MRFASS). Design, methods and baseline characteristics. AB - African-Americans and probably Latinos are at increased risk of stroke compared with white, non-Latino Americans. This study seeks to determine if the known risk factors for stroke can account for this increased risk. In this case-control study controls (neighborhood volunteers) were group-matched to acute stroke cases by ethnicity in a ratio of approximately 2:1 for African-Americans and Latinos and 1:1 for whites. Extensive historical, clinical and laboratory data were collected on each subject. For each ethnic group cases were somewhat older and less well-educated than the volunteer controls. Patients in each ethnic group were similar with regard to time from stroke onset to hospital admission, stroke severity, length of stay, discharge disposition and mortality rate. With minor exceptions the distributions of stroke subtypes within each ethnic group appeared similar to those previously reported. Subject recruitment for this case-control study was completed in the manner and time frame planned. Analysis of risk factor information from this sample should provide valuable information regarding the relative risk associated with the major modifiable risk factors for stroke in the minority groups studied. PMID- 9346343 TI - Risk factors for Alzheimer's disease: a case-control study. AB - The extent of current knowledge of risk factors for Alzheimer's disease (AD) is still limited. We conducted a case-control study of 98 AD patients and 98 age- and sex-matched controls. Light smoking had a decreased risk for AD (AD: 2.0%, controls: 21.4%, odds ratio = 0.10, p = 0.003), whereas daily smoking showed a trend to increase the risk for AD (AD: 45.9%, controls: 26.5%, odds ratio = 1.73, p = 0.08). After multivariable analyses factors associated with AD included the presence of apolipoprotein epsilon4 allele, and the duration of well water consumption. PMID- 9346344 TI - A nationwide epidemiological study of spinal cord injury in geriatric patients in Taiwan. AB - This prospective epidemiological survey of spinal cord injury (SCI) in Taiwan was carried out among patients attended by physicians from various medical centers and general hospitals all over Taiwan from July 1992 to June 1996. In all, 1,586 new cases of SCI were registered, representing about 70% of all possible SCI cases in Taiwan. The observed average annual incidence of SCI in Taiwan was 18.8 per million people, whereas it was 47.5 for the geriatric section. The mean age was 46.1 years with a plateau distribution after 20 years and older. Geriatric victims (297 cases, 18.7%, group II) formed a major section of SCI cases in Taiwan. Another group of younger SCI patients (15-64 years old, 1,232 cases, group I) was selected for comparison. The results showed that the male-to-female ratio, pattern of neurological deficits, and causes of injury and death of geriatric SCI patients differed significantly from those of the younger SCI group. Elderly women were exposed to a higher risk of SCI than younger women (M/F ratio 1.7:1). Falls were the leading cause of geriatric SCI, and two thirds of them occurred on level ground. Traffic accidents accounted for a third of SCI cases, half of which involved motorcycle accidents, a fifth of them pedestrians. Quadriplegia and quadriparesis occurred more frequently among elderly cases of SCI than in the younger group and a higher proportion of them died of SCI complications. Two thirds of elderly SCI patients recovered well enough after comprehensive treatment to be able to take care of themselves at home. The government should initiate programs of prevention to reduce the prevalence of geriatric SCI in Taiwan. PMID- 9346354 TI - Natural killer T cells: a potent cytokine-producing cell population. AB - Natural killer (NK) T cells constitute a subset of TCRalphabeta+ T lymphocytes characterized by natural killer surface receptor expression as well as by a restricted TCR repertoire. This population is capable both of secreting several cytokines, especially IL-4 and IFN-gamma, shortly after stimulation and of killing target cells via Fas/FasL interactions. The cytokines present in the microenvironment of NK T cells play a pivotal role in their potential immunoregulatory functions which are exerted through their ability to induce apoptosis and to modulate the development of Th1 or Th2 cells. Recent reports concerning the physiological roles of the NK T cell population will be discussed in this review. PMID- 9346355 TI - Quantification of cytokine transcripts using polymerase chain reaction. AB - Quantitative PCR techniques are both attractive and daunting. Most users appreciate the need for accurate quantification, but achieving this is not always straightforward. It usually requires more technical equipment than classical PCR and common sense controls are no longer sufficient to generate confidence in the quantitative power of the assay. This review describes the principles that underlie the most commonly used quantitative PCR techniques and their different read-outs, and aims to provide the reader with the tools needed to discriminate between the different published techniques. PMID- 9346345 TI - A study of pediatric brain tumors and their association with epilepsy and anticonvulsant use. AB - OBJECTIVES: To evaluate the risk of childhood brain tumor occurrence in relation to epilepsy and anticonvulsant use. STUDY DESIGN: As part of a multicenter case control study of pediatric brain tumors, maternal report on epilepsy occurrence before diagnosis of her child's brain tumor was collected for 540 cases and compared with 801 control children. Mothers also reported on any long-term (> or = 2 weeks) use of medications by her child before the date of tumor diagnosis (or a comparable reference date for controls) and these medications were classified according to whether they contained barbiturates. RESULTS: As expected, because seizures are often an early brain tumor symptom, a strong association was observed between epilepsy and brain tumor occurrence (odds ratio, OR = 6.2; 95% confidence limit, CL = 2.9, 14). The association remained elevated even after a > or = 10-year interval between diagnoses of epilepsy and brain tumor (OR = 4.7; CL = 0.8, 48). Elevated odds ratios were observed both for epileptic children who were treated with anticonvulsants containing barbiturates (OR = 5.8; CL = 2.2, 18) and for those not treated with barbiturates (OR = 7.9; CL = 1.7, 74), relative to nonepileptic children. CONCLUSION: Whereas most of the brain tumor risk associated with epilepsy may be due to occult tumors, the finding of an elevated risk many years after diagnosis of epilepsy is of interest. PMID- 9346356 TI - Hepatocyte-derived cell lines express a functional receptor for cardiotrophin-1. AB - Cardiotrophin-1 (CT-1) is a recently isolated cytokine belonging to the interleukin-6 cytokine family. In the present study, we show that CT-1 binds to hepatocyte-derived cell lines of rat and human origin with high (Kd = 600-800 pM) and low (Kd approximately 3-6 nM) binding affinities. Treatment of HepG2 cells with CT-1 resulted in the induction of tyrosine phosphorylation of both transducing receptor subunits, gp130 and LIF receptor, and this phosphorylation was completely inhibited by a neutralizing anti-gp130 mAb. Addition of CT-1 to HepG2 or H35 cell cultures induced a dose-dependent production of several acute phase proteins (haptoglobin, fibrinogen, alpha1-acid glycoprotein, alpha2 macroglobulin). Moreover, the use of a neutralizing mAb to gp130 in cultures of HepG2 cells grown in the presence of CT-1, inhibited the induction of acute phase protein secretion, indicating an absolute requirement of gp130 in the formation of a functional CT-1 receptor. Altogether, these results suggest that CT-1 could play an important role in the regulation of hepatocyte metabolism in inflammatory responses. PMID- 9346357 TI - Tumor necrosis factor production capacity as a potentially useful parameter to monitor disease activity in multiple sclerosis. AB - The aim of this study was to develop a test allowing to monitor disease activity in patients with multiple sclerosis (MS). A simple, fast and reliable test was used to assess the cytokine production capacity of blood leucocytes. The whole blood test (WBT) involved in vitro stimulation of a whole blood sample with either the mitogen phytohaemagglutinin (PHA), or specific antigens. We focused our attention on the production of tumor necrosis factor alpha (TNF), because of the possible involvement of this cytokine in MS pathogenesis. Under in vitro stimulation with PHA or MBP, TNF production was found to be significantly higher in patients during the clinical relapses than during remissions. The increment of TNF production correlated with the severity of the relapses, as determined by the modification of Kurtzke EDSS scale. Moreover, each clinical relapse appeared to be preceded by a peak of TNF production. We then retrospectively analysed 21 patients with the relapsing-remitting form of the disease, in whom the WBT was performed every 2-4 weeks, for periods ranging from 16 to 52 months. Seventy three peaks of TNF production (defined as the doubling or more of the individual baseline value, which was found to be stable for each patient during remissions), and 47 relapses, including 36 objective and 11 subjective, were observed. Forty seven out of the 73 TNF peaks were followed by or concomitant with a clinical relapse. In 10 out of the 26 cases where no relapse followed the TNF peak, another cause (mainly infections) of increased TNF production was found. Thus, by excluding other causes, the specificity of the WBT, i.e., the probability to develop a relapse when a TNF peak was found to be 74.6% (47/63). The sensitivity of the WBT was 100%, since all the 47 relapses were preceded by a TNF peak. Assessment of TNF production capacity by the WBT may thus be useful in the follow up of MS patients, particularly for the follow-up of various treatments. Information provided by the WBT may also be useful to orientate the therapeutic decision for an incipient relapse. Earlier treatment is likely to result in an improved prevention of neurological damage. PMID- 9346358 TI - Regression of Morris hepatoma in response to intralesional treatment with tumor necrosis factor muteins. AB - We examined the antitumor effects of human recombinant tumor necrosis factor alpha (rhTNF-alpha) and its muteins with the N-terminal amino acid sequence altered by point mutations against transplantable Morris hepatoma 5123 in rats. In vivo studies showed antiproliferative activity of the drugs in the dose range tested. For in vivo studies rhTNF-alpha and muteins were administered intratumorly (i.t.). The preparations were given at a dose of 10 microg/rat, once daily for eight days. Although the therapy was significantly effective in inhibiting tumor growth, complete growth inhibition could not be achieved. Nevertheless, there was a significant increase in survival time of tumor-bearing rats. PMID- 9346359 TI - Gene transfer-mediated expression of physiological numbers of the type II decoy receptor in a myelomonocytic cellular context dampens the response to interleukin 1. AB - Available information suggests that the type II IL-1 receptor (RII) is a nonsignaling molecule which acts as a decoy for IL-1. The decoy function model for RII was supported by gene transfer experiments in fibroblasts and keratinocytes. Therefore, inhibition of IL-1 responsiveness after decoy RII gene transfer could reflect a non-physiological cellular context and receptor number. In the present study, constructs encoding RII or a cytoplasmic deletion mutant (delta 372-398) were transfected into U937 cells which express only low levels of RI detectable by RT-PCR. Gene transfer resulted in receptor numbers (approximately equal to 10(3)/cell) of the same order of magnitude as that found in normal myelomonocytic cells. Transfer of RII or a cytoplasmic deletion mutant into U937 did not increase responsiveness to IL-1, as assessed by IL-8 expression and production; it actually considerably dampened it. These results are consistent with the view that in a myelomonocytic cellular context, RII does not contribute to signaling and represents a unique pathway of negative regulation of the IL-1 system. PMID- 9346360 TI - Expression of monocyte chemotactic protein-3 in human monocytes and endothelial cells. AB - Monocyte chemotactic protein-3 (MCP-3) is a C-C chemokine which interacts with the CCR1, CCR2 (MCP-1) and CCR3 receptors and has a distinct spectrum of action. The present study was designed to investigate whether human endothelial cells (EC) and, by way of comparison, monocytes express MCP-3 and its regulation by pro and anti-inflammatory cytokines. Unstimulated human umbilical vein EC and monocytes did not express MCP-3. Bacterial lipopolysaccharides (LPS), IL-1 and TNF induced low, but appreciable levels of MCP-3 transcripts. Interferon-gamma was a much weaker, but nevertheless active, inducer. MCP-3 transcripts were considerably less abundant than those for MCP-1. IL-4, IL-13 and IL-10 did not induce MCP-3 expression. These anti-inflammatory cytokines suppressed cytokine induced MCP-3 expression in monocytes. In contrast, IL-4, IL-13 and IL-10 either did not affect or they amplified MCP-3 induction in EC. EC-produced MCP-3 may be important in the regulation of extravasation of receptor-expressing cells, including NK cells and eosinophils. PMID- 9346361 TI - Host's immune response in electrotherapy of murine tumors by direct current. AB - Electrotherapy by low level direct current has been demonstrated to have antitumor effects in different murine tumor models and in clinics. Electrotherapy in "field" configuration, where electrodes are placed subcutaneously outside of the tumor in a way that tumor lies in between the electrodes, was performed in immunodeficient nude and immunocompetent mice. Electrotherapy was much more effective in immunocompetent mice based on the observed tumor growth retardation, thus demonstrating that antitumor effectiveness of electrotherapy greatly depends on host's immune response. Further experiments were conducted by combining electrotherapy with concomitant immunotherapy in order to potentiate the antitumor effect of electrotherapy. Immunotherapy consisted of local delivery of genetically engineered cells selected for IL-2 secretion. This combined treatment was much more effective than any of the treatments alone. PMID- 9346362 TI - Circulating interleukin-6 type cytokines in patients with systemic lupus erythematosus. AB - We investigated the serum concentration of interleukin-6 (IL-6) and the IL-6 family of cytokines (leukemia inhibitory factor (LIF), oncostatin M (OSM) and ciliary neurotrophic factor (CNTF) using an enzyme-linked immunosorbent assay (ELISA) in 64 patients with systemic lupus erythematosus (SLE) and 15 healthy controls. We also examined a possible association between the serum levels of these proteins and SLE activity as well as correlations between the IL-6 concentration and the levels of LIF, OSM and CNTF. IL-6 was detectable in all 64 patients with SLE and normal individuals, and the level of this cytokine was significantly higher in patients than in the control group (p < 0.002 ). LIF, OSM and CNTF were detectable in 9 (14.1%), 6 (9.4%) and 51 (78%) patients, respectively, and undetectable in the majority of healthy individuals. We found positive correlation between the serum concentrations of IL-6, LIF, OSM and CNTF and SLE activity. IL-6 and OSM serum levels were also correlated but not IL-6 and LIF or CNTF. In conclusion, an increase in the serum levels of IL-6 and, to a lesser extent of LIF, OSM and CNTF concentrations may be useful markers for SLE activity. PMID- 9346363 TI - Cytokines: a pharmacologically-oriented introduction. PMID- 9346364 TI - Clinical pharmacology of cytokines. PMID- 9346365 TI - Blocking interleukin-1 and tumor necrosis factor in disease. PMID- 9346366 TI - Anti-tumor necrosis factor alpha therapy of rheumatoid arthritis. Mechanism of action. PMID- 9346367 TI - Interleukin-10 as an anti-stress cytokine. PMID- 9346368 TI - Clinical development of interleukin-10. PMID- 9346369 TI - Biological properties and therapeutic applications of interleukin-12. PMID- 9346370 TI - Erythropoietin: from molecular biology to clinical use. PMID- 9346371 TI - Thrombopoietin (Mpl ligand) and the regulation of platelet production. PMID- 9346372 TI - Immunointervention based on Th2-type cytokines in HIV infection. PMID- 9346373 TI - Cytokines in severe infections: from pathophysiology to clinical applications. PMID- 9346374 TI - Interferon alpha in hepatitis C: a cytokine for reducing fibrosis progression. PMID- 9346376 TI - GP130 cytokines, interferon, survival and proliferation of human myeloma cells. PMID- 9346375 TI - Cytokines as prognostic and therapeutic tools in human cancer. PMID- 9346377 TI - The dream of effective cytokine-based tumor vaccines. PMID- 9346379 TI - Gene transfer into the kidney: current status and limitations. AB - Gene therapy is obviously a controversial issue and a wave of suspicion has dampened the initial enthusiasm raised by this new therapeutic approach. It has now become fashionable to downplay the potential for gene therapy in most fields including kidney-related diseases. In our opinion, this is an unfair and unrealistic view of the future. In fact, gene therapy of well-selected kidney diseases will certainly become feasible, but a large data base on vectors and transfer methods both in the normal kidney and in disease models has first to be collected. Any significant progress in the biology of the vectors, in the cellular interactions of the newly introduced DNA, and in the regulation and persistency of the transgene should be rapidly translated to the kidney in relevant experimental models. Herein, we present the use and current limitations of gene transfer to the kidney and the potential therapeutic perspectives. PMID- 9346378 TI - The validity of lithium clearance as an index of sodium and water delivery from the proximal tubules. PMID- 9346380 TI - Exercise renal rehabilitation program: psychosocial effects. AB - The aim of this study was to assess the psychosocial effects of exercise training on hemodialysis (HD) patients. Thirty-one uremic patients, aged 50.6+/-11.6 years, on maintenance HD were studied. Twenty patients were selected at random for a 6-month exercise renal rehabilitation program (ERRP) consisting of 3 weekly sessions of exercise training. The other 11 patients were assigned to sedentary control status. A formal psychosocial assessment, which included affective (Beck Depression Inventory, BDI), quality of life (Quality of Life Index, QLI) and personality (Eysenck Personality Questionnaire, EPQ) parameters, was performed with validated questionnaires at the beginning and the end of the ERRP. After training significant improvement occurred in physical capacity (VO2max increased from 16.8+/-6.2 to 23.2+/-7.6 ml/kg/min, p < 0.05). Although the level of depression did not differ betwen the 2 groups at pretesting, the ERRP group showed a decrease in their self-report of depression (decrease in BDI score value, from 21.0+/-10.4 to 13.7+/-9.5, p < 0.05) after the training program. From the relationship between the baseline levels of BDI depression and changes in VO2max in the ERRP group it was suggested that the most severely depressed patients got the greatest beneficial effects from exercise training. Moreover, trained patients demonstrated an improvement in QLI (from 6.3+/-1.5 to 9.0+/-0.9, p < 0.05). This improvement was found to be dependent on the participation in ERRP, the effects of the training and the improvement in the depression. All the above functional and psychosocial parameters remained unchanged in the controls. The results demonstrate that ERRP is an effective emotional therapeutic method for HD patients and improves their quality of life. PMID- 9346381 TI - Continuous ambulatory peritoneal dialysis in patients after intra-abdominal prosthetic vascular graft surgery. AB - The purpose of this study was to assess the feasibility of continuous ambulatory peritoneal dialysis (CAPD) after intra-abdominal prosthetic vascular graft surgery. We report 8 consecutive patients with end-stage renal disease, who previously underwent intra-abdominal prosthetic aortic graft replacement, treated by CAPD between November 1983 and November 1994. All patients received a peritoneal dialysis catheter without technical problems and were dialyzed for a total of 208 months. Six episodes of peritonitis occurred in 4 patients without clinical evidence of any abdominal aortic graft infection. Three patients developed intermittent claudication and 2 died of myocardial infarct. A similar peritonitis and cardiovascular complication rate was observed in a control group of age- and sex-matched CAPD patients with no aortic prosthesis. We conclude that CAPD is feasible in patients with abdominal aortic prosthesis. PMID- 9346382 TI - Angiotensin-converting enzyme inhibitors are associated with the need for increased recombinant human erythropoietin maintenance doses in hemodialysis patients. Risks of Cardiac Disease in Dialysis Patients Study Group. AB - The influence of angiotensin-converting enzyme inhibitors (ACEIs) on recombinant human erythropoietin (rhEPO) maintenance doses in hemodialysis patients was studied. One hundred and eight chronic hemodialysis patients (55 males and 53 females, mean age 61.2+/-12.6 years) were investigated. The rhEPO maintenance doses in the ACEI-treated group (n = 49) were 101.7+/-51.7 U/kg/week and in the nontreated group (n = 59) 79.2+/-37.8 U/kg/week (p < 0.05). No difference was observed in hematocrit between the ACEI-treated and nontreated groups. In stepwise regression analysis, the parameters associated with increased rhEPO maintenance doses were female gender, ACEI administration, low total iron binding capacity, and low serum free carnitine levels. In conclusion, ACEI administration might reduce the response to rhEPO. In hemodialysis patients who need high-dose rhEPO to maintain the target hematocrit in the absence of iron deficiency, hyperparathyroidism, infection, malignancy, malnutrition, and aluminum toxicity, ACEI administration should be considered. PMID- 9346384 TI - Reversal of anemia by erythropoietin therapy retards the progression of chronic renal failure, especially in nondiabetic patients. AB - Therapy with human recombinant erythropoietin (EPO) has been accepted as effective for renal anemia in dialysis patients. However, studies in rats have shown that correcting anemia with EPO may affect the progression of renal dysfunction. In humans, however, the effect of EPO on residual renal function is a matter of controversy. We, therefore, investigated whether the long-term administration of EPO to predialysis patients influences residual renal function. Anemic patients at the predialysis stage with a serum creatinine (Cr) concentration ranging from 2 to 4 (average 2.9) mg/dl and a hematocrit (Ht) of less than 30% were randomly assigned to two groups which consisted of anemic patients not treated with EPO (group I, untreated anemic controls, n = 31) and anemic patients treated with EPO (group II, treated anemics, n = 42). Patients with nonsevere or moderate anemia (Ht > 30%) with a Cr ranging from 2 to 4 (average 2.6) mg/dl were also recruited as nonanemic controls (group III, untreated nonanemic controls, n = 35). Blood pressure was controlled to the same degree among the three groups by combined treatment with calcium antagonists and angiotensin-converting enzyme inhibitors. All patients were kept strictly on a low-protein (0.6 g/kg/day) and a low-salt (7 g/day) diet. The degree of control of dietary protein and blood pressure and the frequency of angiotensin-converting enzyme inhibitor administration were comparable among the three groups. The primary end point for each patient was a doubling of the baseline Cr which yielded cumulative renal survival rates which were plotted against time. Ht rose significantly from 27.0+/-2.3 to 32.1+/-3.2% in group II (n = 42, p < 0.001) with a rate of increase of 0.4+/-0.06%/week. However, it declined from 27.9+/-1.8 to 25.3+/-1.9% in group I (n = 31, p < 0.001) and from 35.9+/-3.5 to 32.2+/-3.9% in group III (n = 35, p < 0.001). Cr doubled in 26 patients (84%) in group I as compared with 22 (52%) in group II and 21 (60%) in group III. The cumulative renal survival rates in groups II and III were significantly better than that in group I: p = 0.0003 (group I vs. group II) and p = 0.0024 (group I vs. group III). However, there was no difference in the renal survival rate between groups II and III (p = 0.3111). The better survival rate obtained in group II was attributable to the better survival rate for the nondiabetic patients in this group. The present study suggests that anemia, per se, is a factor in the progression of end-stage renal failure and that reversal of anemia by EPO can retard the progression of renal failure, especially in nondiabetic patients, provided that blood pressure control, rate of increase in Ht, and dietary protein restriction are appropriate. PMID- 9346383 TI - Effects of erythropoietin on gonadotropin responses to gonadotropin-releasing hormone in uremic patients. AB - Long-term therapy with recombinant human erythropoietin (rhEPO) in uremic male patients undergoing hemodialysis has been followed by an increase in plasma levels of testosterone and a decrease in baseline levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). The aim of the present study was to assess the effect of acutely administered rhEPO on FSH and LH responses to gonadotropin-releasing hormone (GnRH) in a group of uremic patients undergoing continuous ambulatory peritoneal dialysis (CAPD). Sixteen clinically stable male patients (age, mean+/-SEM, 45.3+/-3.9 years) with chronic renal insufficiency and 12 healthy volunteers with a normal renal function, matched for age and body mass index, were studied. All patients were on CAPD therapy for at least 3 months, and none of them received rhEPO therapy. Patients were moderately anemic (hemoglobin 11.0+/-0.3 g/dl) and showed testosterone levels significantly lower than those found in control subjects (3.47+/-0.37 vs. 6.91+/-0.49 ng/ml, p < 0.001). Each subject was tested with GnRH (100 microg i.v. as bolus) and with GnRH plus rhEPO (40 U/kg at a constant infusion rate for 30 min, starting 15 min before GnRH injection) on different days. Blood samples for FSH and LH were obtained between 30 and 120 min. In uremic patients the baseline FSH levels were higher than those found in control subjects (18.88+/-5.41 vs. 6.41+/-1.10 mU/ml, p < 0.05). After GnRH administration FSH values reached a maximum of 25.50+/-6.19 mU/ml in patients and of 12.50+/-2.02 mU/ml in controls (p < 0.05). rhEPO infusion produced a significant (p < 0.01) decrease in the area above the baseline value of FSH in uremic patients, with no other change in FSH responses to GnRH both in patients and controls. Baseline LH concentrations were significantly higher in patients than in controls (15.56+/-3.41 vs. 2.58+/-0.36 mU/ml, p < 0.001). LH peak and area under the curve of LH secretion after GnRH were significantly higher in patients than in controls (45.25+/-6.28 vs. 26.83+/-4.62 mU/ml, p < 0.05, and 77.02+/-11.30 vs. 34.40+/-5.22 mU x h/ml, p < 0.005, respectively). When GnRH was injected during the rhEPO infusion, a significant (p < 0.02) reduction in LH concentrations at 60, 90, and 120 min was found in uremic patients. Accordingly, the LH area under the curve was significantly reduced in patients (65.99+/-11.44 mU x h/ml, p < 0.05). rhEPO had no effect on GnRH-induced LH release in control subjects. These results suggest that acute rhEPO administration might reduce the exaggerated LH response to GnRH stimulation found in uremic male patients on CAPD. PMID- 9346385 TI - Treatment of acute rejection in live related renal allograft recipients: a comparison of three different protocols. AB - We present our experience on the comparison of three different modes of steroid therapy, oral prednisolone (OP), intravenous dexamethasone (IVDX) and intravenous methylprednisolone (IVMP) in the treatment of acute rejection (AR) in renal allograft recipients. Between January 1980 and January 1992, 206 patients underwent live related renal transplantation. Before 1990, all received prednisolone (PRED) and azathioprine (AZA) only. After 1990, patients were given PRED, AZA and cyclosporine (CsA). After 1 year, CsA was stopped and patients were converted to a two-drug regimen only. Of the 206 patients, 180 (87.4%) were male and mean age was 30.3+/-8.7 years (range 14-63). During the mean follow-up of 43.5 months, 178 episodes of AR were seen in 121 patients. Each episode was considered as a separate entrant in the study. Conventional immunosuppression was given in 151 episodes and 27 episodes were on triple-drug therapy. Diagnosis of AR was made by clinical, sonography, nuclear scan with or without graft biopsy evidence. Of the 178 AR, 110 (61.8%) were within 3 months, 36 (20.2%) were between 3 months and 1 year and 32 (18%) were after 1 year. OP was given in 11 cases while IVDX and IVMP were given in 48 and 119 cases respectively. Overall, 154 (86%) showed either a complete or partial response to antirejection therapy. Response to therapy was 91, 90 and 85% in OP, IVDX and IVMP groups respectively. There was no statistical difference in response rate in different groups. There was also no difference in side effects in three different groups. Our data suggest that it is the high dose of steroid rather than mode of therapy which is responsible for therapeutic benefit in treatment of AR. PMID- 9346387 TI - Effect of angiotensin II on plasminogen activator inhibitor-1 production by cultured human mesangial cells. AB - Angiotensin II is a vasoactive peptide that has been widely implicated in the pathogenesis of glomerular disease. Some of its effects are thought to be independent of changes in blood pressure. Plasmin is a key regulator of fibrinolysis and extracellular matrix turnover. The conversion of plasminogen to plasmin by plasminogen activators (PAs) is controlled by their specific inhibitor, PAI-1. In this study we report the effects of angiotensin II on the production of PA inhibitor-1 (PAI-1) and tissue-type PA (t-PA) by glomerular mesangial cells in culture. Angiotensin II significantly increased the production of PAI-1 in the supernatant of mesangial cells (p < 0.05) in a dose-dependent manner, the maximum stimulation occurring at a concentration of 10(-5) M. The effect was not mediated by transforming growth factor-beta (TGF-beta), which is known to be induced by angiotensin II; TGF-beta itself can increase PAI-1 expression. Angiotensin II did not alter t-PA production or incorporation of matrix fibronectin but did increase cellular proliferation and 3H-thymidine uptake. The increase in PAI-1 by angiotensin II may contribute to the persistence of fibrin deposits and extracellular matrix accumulation, providing another mechanism whereby angiotensin II contributes to glomerular dysfunction. PMID- 9346386 TI - Molecular epidemiology of hepatitis C virus infection in dialysis patients. AB - There are very few data on the molecular biology of hepatitis C virus (HCV) infection in dialysis patients. 101 patients undergoing dialysis treatment in 4 units in the Lombardy, northern Italy, were analyzed by RT-PCR for HCV viremia, by line probe assay technology for HCV genotyping and by a serological analysis for detecting type-specific antibodies. 61 of 101 (60%) patients showed detectable HCV RNA in serum; HCV genotype 2a was dominant (30/53 = 57%), followed by HCV genotype 1b (20/53 = 37%). There was no relationship between HCV genotyping and the clinical or demographic features of the patients. The antibody response toward the c33-c, c100-3, and 5-1-1 antigens was more frequent in HCV genotype 1b compared with genotype 2a (p = 0.046, p = 0.001 and p = 0.0001, respectively). The antibody levels to NS-3 and NS-4 HCV proteins were significantly higher in patients with-HCV genotype 1b in comparison with HCV 2a infected individuals (p = 0.0001). There was a high level (82%) of agreement between HCV genotyping by RT-PCR and the assessment of type-specific antibodies by serological analysis; further, it was possible to detect type-specific antibodies in 6 of 22 (27%) patients in whom PCR amplification was unsuccessful. In conclusion, HCV subtype 2a was dominant in our population of HCV-infected dialysis patients, dialysis patients infected by different genotypes showed similar demographic and clinical characteristics, the antibody response toward the NS-3- and NS-4-related antigen of HCV was genotype dependent. There was a high level of agreement between HCV genotyping by RT-PCR and the detection of type-specific antibodies by serological analysis. As significant biological differences may exist among HCV strains, the assessment of HCV types may be very useful in the routine clinical activity of nephrologists in dialysis units. PMID- 9346388 TI - Aminoglycoside-associated Fanconi's syndrome: an underrecognized entity. AB - The Fanconi syndrome is an array of multiple proximal renal tubular dysfunctions occurring in association with several exogenous toxins, such as aminoglycosides. These antibiotics remain the drugs of choice in most gram-negative infections, but nephrotoxicity is the main drawback for them. Furthermore, the nephrotoxic effects may be overlooked with routine analyses. With the purpose of making physicians aware of this underrecognized complication, we are reporting here 3 cases of Fanconi's syndrome related to the administration of high-dose aminoglycosides (ranging from 3.6 to 15 g) with normal serum urea nitrogen and creatinine levels. The pattern of aminoaciduria demonstrated high increases in neutral amino acids, followed by dibasic and near-normal acidic amino acids. We also report the urinary excretion rates of total protein and beta2-microglobulin and protein electrophoresis results. We compared these cases with others reported in the literature. PMID- 9346389 TI - Interleukin 10 and interleukin 13 synergize to inhibit vascular permeability factor release by peripheral blood mononuclear cells from patients with lipoid nephrosis. AB - It has been proposed that a vascular permeability factor (VPF) is involved in the pathogenesis of lipoid nephrosis (LN). There is now increasing evidence that interleukin 10 (IL-10) and interleukin 13 (IL-13) have regulatory effects on cytokine production by activated macrophages. These results prompted us to study the effects of recombinant human IL-10 and IL-13 on VPF secretion in LN. In the present study, we demonstrate that the regulatory cytokines IL-10 and IL-13 are potent inhibitors of the VPF activity of activated peripheral blood mononuclear cells. Each cytokine was found to suppress VPF secretion in a dose-dependent fashion. More importantly, the combination of the cytokines was found to give a potent synergistic suppression of VPF by concanavalin A activated peripheral blood mononuclear cells from patients with LN. When both anti-IL-10 and anti-IL 13 antibodies were added together to the peripheral blood mononuclear cells, a further increase of concanavalin A enhanced secretion of VPF occurred. These data establish IL-10 and IL-13 as potent inhibitors of VPF activity and suggest their utility in controlling deleterious VPF-mediated responses such as occur in LN patients with nephrotic syndrome. PMID- 9346390 TI - Influence of heparin and type-IV collagen on IL-6 synthesis by rat glomerular mesangial cells. AB - Recent studies have revealed the potential importance of the extracellular matrix (ECM) in the modulation of mesangial cell (MC) function. Interleukin-6 (IL-6) is produced by MCs and was shown to induce MC proliferation, acting as an autocrine growth factor. Heparin is a known inhibitor of MC proliferation. It was shown to modulate ECM synthesis by cultured MCs. The action of heparin on IL-6 synthesis by MCs is presently unknown. We investigated the effect of heparin on IL-6 production when MCs were cultured with or without type-IV collagen, a major constituent of ECM. When MCs were cultured without coating, heparin significantly decreased their IL-6 production; on type-IV collagen, heparin had no significant effect. When tumor necrosis factor alpha (TNF-alpha) was used to stimulate the cells to produce IL-6, heparin was able to decrease the stimulatory effect of TNF alpha when the cells were cultured on plastic but not when in contact with type IV collagen. Thus we conclude that heparin has an inhibitory effect on IL-6 secretion by MCs that is prevented by type-IV collagen. PMID- 9346391 TI - Effect of morphine on renomedullary interstitial cell proliferation and matrix accumulation. AB - Renal interstitial scarring is an important feature of heroin-associated nephropathy. We studied the effect of morphine, an active metabolite of heroin, on cultured rat renal medullary interstitial cell (RMIC) proliferation and matrix accumulation. Morphine (10(-12) M) enhanced (p < 0.001) the proliferation of RMIC (control, 15.0+/-0.5 vs. morphine, 20.4+/-1.1 x 10(4) cells/ml). This effect of morphine was dose and time dependent. [3H]thymidine and bromodeoxyuridine incorporation studies confirmed the mitogenic effect of morphine on RMIC. Morphine also enhanced mRNA expression for c-jun and c-myc on RMIC. However, nalbuphine, a non-addicting alkaloid did not modulate the proliferation of RMIC. Morphine enhanced the accumulation of collagen type I in a dose-dependent manner and also increased (p < 0.001) the accumulation of collagen type III at a high concentration (control, 1,291+/-55.8 vs. morphine, 10(-4) M, 2,697.6+/-257.8 ng/microg protein). Morphine did not modulate the accumulation of laminin or fibronectin. Neutralizing antibody to IL-6 inhibited the effect of morphine on RMIC. H7, a protein kinase C inhibitor, also attenuated the morphine-induced RMIC proliferation. The present study provides a basis for a hypothesis that morphine may be playing a role in the development of renal interstitial pathology in patients with heroin addiction. PMID- 9346392 TI - Alpha-1-antitrypsin deficiency associated with hepatic cirrhosis and IgA nephritis. AB - The association between alpha1-antitrypsin (A1AT) deficiency and glomerulonephritis has only sporadically been reported, and mostly based upon autopsy findings, as opposed to the more frequent linkage between A1AT deficiency and lung emphysema with or without hepatic cirrhosis. The present case report describes a 30-year-old man with A1AT deficiency, without evidence of lung disease, who developed hepatic cirrhosis in early childhood and IgA glomerulonephritis and hypertension in adult life. The IgA nephritis followed an unusual course, with a sudden deterioration of the renal function, possibly induced by uncontrolled hypertension or the possible occurrence of vasculitis. After 6 months of hemodialysis, the patient successfully underwent living-related donor kidney transplantation. PMID- 9346393 TI - Reactivation of systemic lupus erythematosus by pregnancy in a hemodialysis patient. PMID- 9346395 TI - Measurement of cortical bone density in the radius by peripheral quantitative computed tomography in hemodialysis patients. PMID- 9346394 TI - Treatment of severe combined overdose of calcium antagonists and converting enzyme inhibitors with angiotensin II. PMID- 9346396 TI - Carbohydrate diet prolongs survival of rats with acute uremia after bilateral nephrectomy. PMID- 9346397 TI - Development of oxidative stress in chronic kidney insufficiency following the progression of disease. PMID- 9346398 TI - Detection of renal artery stenosis with doxazosin. PMID- 9346400 TI - The influence of hyperthyroidism on vasoconstrictor and vasodilator responses in isolated coronary and renal resistance arteries. AB - The influence of hyperthyroidism on the functional vascular responsiveness of isolated coronary and renal resistance vessels was investigated. Hyperthyroidism was established by feeding rats for 1 and 4 weeks with 5 mg/kg L-thyroxine (T4) containing rat chow. Preparations of either coronary or renal resistance vessels were mounted in an isometric wire myograph. Subsequently, concentration-effect curves were determined for the effects of 5-hydroxytryptamine (5-HT), 9,11 dideoxy-11alpha,9alpha-epoxymethanoprostaglandin F2alpha (U46619) and isoproterenol in coronary vessels, and for those of methoxamine, U46619 and isoproterenol in renal vessel preparations. Our results indicate that hyperthyroidism does not induce major changes in the sensitivity of both coronary and renal resistance vessels towards 5-HT, U46619 and methoxamine. A clearly sensitizing influence of acute hyperthyroidism (1 week of T4 treatment) was found for isoproterenol-induced relaxant responses, whereas hyperthyroidism for 4 weeks did not influence the responses mediated by isoproterenol in coronary resistance arteries. Furthermore, the isoproterenol-induced relaxation in renal arteries was not influenced by the chronic hyperthyroid state of the animal. The present results indicate that in acute hyperthyroidism beta-adrenoceptor-mediated vasodilation is increased. However, in chronic hyperthyroidism changes in responsiveness to vasoconstrictor or vasodilator agents of coronary and renal resistance arteries appear not to play a major role. The influence of hyperthyroidism on the functional response of resistance arteries appears to be both tissue and time dependent. PMID- 9346399 TI - Effects of L-DOPA/carbidopa administration on the levels of L-DOPA, other amino acids and related compounds in the plasma, brain and heart of the rat. AB - Rats were treated intraperitoneally with a mixture of 250 mg/kg L-DOPA and 40 mg/kg carbidopa or with vehicle and sacrificed 30 min later. Plasma, heart and cortex, midbrain, brainstem and cerebellum were removed from each animal and assayed by HPLC for L-DOPA and a large number of amino acids and related amino compounds. L-DOPA levels increased from undetectable (<0.2 nmol/ml or g) to 1,146, 1,007, 399, 376, 368 and 850 nmol/ml or g in the above tissues. In addition, several amino compounds were significantly affected by L-DOPA/carbidopa (p < or = 0.01). Plasma concentrations of phosphoserine, oxidized glutathione, citrulline, phenylalanine, tyrosine and 1-methylhistidine increased and arginine, glutamic acid and lysine decreased. In the heart, concentrations of phosphoserine, taurine, reduced glutathione, threonine, serine, glutamine, glycine, alanine, valine, GABA, ethanolamine, ammonia and arginine decreased. In the cortex, camosine and homocarnosine increased. In the midbrain, valine increased and leucine, ornithine and oxidized glutathione decreased. In the cerebellum, citrulline increased. In the brainstem, threonine, serine, asparagine, glutamine, oxidized glutathione, alanine, and leucine decreased. In the brainstem, arginine was slightly decreased with a concomitant increase in citrulline (p < 0.05), indicative of nitrous oxide formation. These results show that administration of L-DOPA/ carbidopa not only raises dopamine levels but can also affect other biochemicals and that the observed changes in amino acids and related compounds can perhaps contribute to the beneficial and/or adverse effects of L-DOPA/carbidopa therapy of Parkinson's disease. PMID- 9346401 TI - Effects of aminoguanidine on adhesion molecule expression of human endothelial cells. AB - The effect of aminoguanidine (AG) on the expression of adhesion molecules on nonactivated human umbilical vein endothelial cells (HUVEC) was investigated in vitro. Nonactivated HUVEC cultivated on long-term glycated fibronectin (FN) as compared to native FN showed a significant upregulation of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and CD31 which could be further promoted by long-term glycated bovine serum albumin. AG, at a concentration of 0.01 mol/l, caused an upregulation of ICAM-1 of 48 +/- 17.4% in HUVEC cultivated on gelatin. In contrast, VCAM-1 and E-selectin remained unaffected. At this concentration, formation of advanced glycation end products (AGE) was inhibited by 57%, as determined immunologically, and by 50%, as verified by AGE-specific fluorescence. A hypothesis concerning the upregulation of ICAM-1 by AG as compared to VCAM-1 is proposed relating to its relative redox insensitivity. Our results demonstrate that the beneficial effect of AG in reducing the risk of accelerated development of atherosclerosis in diabetic patients by inhibiting formation of AGE on matrix proteins such as FN might be hampered by its tendency to upregulate ICAM-1 on endothelial cells. PMID- 9346402 TI - Subcellular distribution of SERCA and calcium-activated ATPase in rabbit and human urinary bladder smooth muscle. AB - Previous studies have demonstrated that calcium storage and release from IP-3 dependent sites in the sarcoplasmic reticulum play an important role in the contractile response of the rabbit urinary bladder to both field stimulation (mediated via neurotransmitter release) and bethanechol (direct muscarinic stimulation). In view of the importance of SERCA in urinary bladder smooth muscle function, we studied the distribution of SERCA by two methods: using Western blotting to quantitate the protein concentration and by enzyme analysis using thapsigargin to specifically inhibit SERCA. Rabbit and human samples of urinary bladder smooth muscle were homogenized and the homogenate separated into three particulate fractions by differential centrifugation: nuclear-cell wall, mitochondrial, and microsomal. The protein concentration of these three particulate fractions was determined and the SERCA protein level quantitated by Western blotting using SERCA-2 antibodies. The calcium-ATPase activity was quantitated using standard enzymatic analysis and the thapsigargin sensitivity determined. The results demonstrated that: (1) the concentration of SERCA was significantly greater in the microsomal fraction than in either of the other fractions for both rabbit and human bladder smooth muscle; (2) the enzymatic activities of both total calcium-activated ATPase and thapsigargin-sensitive calcium ATPase were evenly divided among the three fractions, and (3) the enzymatic activity of both total calcium-activated ATPase and thapsigargin sensitive calcium ATPase of the rabbit exceeded that of the human. In conclusion, the distribution of SERCA and calcium-ATPase of the rabbit bladder smooth muscle was similar to that in the human bladder smooth muscle, although activities in rabbit were significantly greater than those of human tissue. PMID- 9346403 TI - Effects of DQ-2511, a novel prokinetic agent, on electrical activities of smooth muscle in the guinea pig stomach. AB - Electrophysiological experiments were carried out to investigate the prokinetic actions of DQ-2511 on isolated smooth muscle of the guinea pig stomach. DQ-2511 enhanced the myogenic gastric slow waves and cholinergic excitatory junction potential and reduced the frequency of slow waves and the nonadrenergic, noncholinergic, and nonnitrergic inhibitory junction potential, with no significant alteration of the resting membrane potential. The results suggest that the prokinetic actions of DQ-2511 involve excitatory actions directly on smooth muscle and indirectly on cholinergic transmission. PMID- 9346404 TI - Effects of cimetidine on acute gastric mucosal injury induced by ischemia reperfusion in rats. AB - We investigated the effects of cimetidine on acute gastric mucosal injury induced by ischemia-reperfusion in rats. Under pentobarbital anesthesia, the celiac artery was clamped for 30 min and reperfused for 60 min. Cimetidine, famotidine and omeprazole caused a dose-dependent suppression in the total area of erosions that were induced by ischemia-reperfusion. Whereas, none of them inhibited the increase in thiobarbituric acid-reactive substances in the stomach, as an index of lipid peroxidation. The inhibitory effect of intraperitoneally administered cimetidine on mucosal damage was abolished by continuous luminal perfusion with HCl solution (pH 1.5, 1 ml/min) during ischemia-reperfusion, while luminal perfusion with the solution containing HCl and cimetidine (3 mmol/l) significantly reduced the total area of erosions compared to luminal perfusion with HCl solution alone. Cimetidine (3 mmol/l) inhibited hydroxyl radical-induced lipid peroxidation of human erythrocyte membranes by 60% in vitro. These results indicate that cimetidine possesses a protective effect against acute gastric mucosal injury induced by ischemia-reperfusion not only due to the suppression in gastric acid secretion, but also due to the antioxidant action when it is present at a high concentration in the intragastric environment. PMID- 9346425 TI - Oxidative stress in closed-head injury: brain antioxidant capacity as an indicator of functional outcome. AB - It has been suggested that reactive oxygen species (ROS) play a role in the pathophysiology of brain damage. A number of therapeutic approaches, based on scavenging these radicals, have been attempted both in experimental models and in the clinical setting. In an experimental rat and mouse model of closed-head injury (CHI), we have studied the total tissue nonenzymatic antioxidant capacity to combat ROS. A major mechanism for neutralizing ROS uses endogenous low molecular weight antioxidants (LMWA). This review deals with the source and nature of ROS in the brain, along with the endogenous defense mechanisms that fight ROS. Special emphasis is placed on LMWA such as ascorbate, urate, tocopherol, lipoic acid, and histidine-related compounds. A novel electrochemical method, using cyclic voltammetry for the determination of total tissue LMWA, is described. The temporal changes in brain LMWA after CHI, as part of the response of the tissue to high ROS levels, and the correlation between the ability of the brain to elevate LMWA and clinical outcome are addressed. We relate to the beneficial effects observed in heat-acclimated rats and the detrimental effects of injury found in apolipoprotein E-deficient mice. Finally, we summarize the effects of cerebroprotective pharmacological agents including the iron chelator desferal, superoxide dismutase, a stable radical from the nitroxide family, and HU-211, a nonpsychotoropic cannabinoid with antioxidant properties. We conclude that ROS play a key role in the pathophysiology of brain injury, and that their neutralization by endogenous or exogenous antioxidants has a protective effect. It is suggested, therefore, that the brain responds to ROS by increasing LMWA, and that the degree of this response is correlated with clinical recovery. The greater the response, the more favorable the outcome. PMID- 9346426 TI - Noninvasive quantitative measurements of regional cerebral blood flow using technetium-99m-L,L-ECD SPECT activated with acetazolamide: quantification analysis by equal-volume-split 99mTc-ECD consecutive SPECT method. AB - Resting- and acetazolamide (Acz)-activated-regional cerebral blood flow (rCBF) measurements were performed by consecutive single-photon emission computed tomography (SPECT) studies before and after Acz administration using equal-volume split technetium-99m-L,L-ethyl cysteinate dimer. Quantitative rCBF images were converted from qualitative axial SPECT images by the application of Patlak plot graphical analysis with radionuclide angiography and Lassen's linearization correction. Total time span required for this study was 53 minutes. The unaffected side of 37 studies with unilateral vascular lesions and 45 studies without apparent vascular lesions showed 132 +/- 17% and 140 +/- 15% increase of mean CBF (mCBF), respectively, under Acz administration. Comparing these values, the Acz-activated rCBF increases of less-affected and affected hemispheres of 23 studies with bilateral vascular lesions (116 +/- 13% and 113 +/- 12%, respectively) was lower with high statistical significance (P < 0.001). For the other 20 cases, physiologic saline was administered instead of Acz. This group showed no changes in mCBF under placebo administration (after placebo/baseline; 100 +/- 6%). Acetazolamide-activated rCBF increase was recognized clearly and easily using quantitative images. This noninvasive method is easy to perform and may be helpful to detect regional abnormalities of hemodynamic reserve in cerebrovascular diseases. PMID- 9346427 TI - Performance of a randomization test for single-subject (15)O-water PET activation studies. AB - We adapted and implemented a permutation test (Holmes 1994) to single-subject positron emission tomography (PET) activation studies with multiple replications of conditions. That test determines the experimentwise alpha error as well as location and extent of focal activations in each individual. Its performance was assessed in five normal volunteers, using (15)O-H2O-PET data acquired on a high resolution scanner, with septa retracted (3D mode), during functional activation by repeating words versus resting (four replications each). Calculated alpha errors decreased and the size of activated tissue volumes (voxels with P < or = 0.05) increased with increasing filter kernel size applied to the difference images. At a filter kernel of 12 mm Gaussian full width at half maximum, significant focal activations were seen bilaterally in superior temporal cortex, including Brodmann's areas 41 and 42, in all five subjects. Additional foci were detected in the precentral gyrus, left inferior frontal gyrus, supplementary motor area, and cerebellum of several subjects. The average CBF increase in activated voxels ranged from 17.6% to 28.7%. Activated volumes were smaller than those detected with a standard parametric test procedure. We conclude that the permutation test is a less sensitive procedure, having the advantage of not depending on unproven distributional assumptions, that detects strong activation foci in individual subjects with high reproducibility. PMID- 9346428 TI - Oxidative glucose metabolism in rat brain during single forepaw stimulation: a spatially localized 1H[13C] nuclear magnetic resonance study. AB - In the alpha-chloralose-anesthetized rat during single forepaw stimulation, a spatially localized 1H[13C] nuclear magnetic resonance spectroscopic method was used to measure the rate of cerebral [C4]-glutamate isotopic turnover from infused [1,6-(13)C]glucose. The glutamate turnover data were analyzed using a mathematical model of cerebral glucose metabolism to evaluate the tricarboxylic acid (TCA) cycle flux (V(TCA)). During stimulation the value of V(TCA) in the sensorimotor region increased from 0.47 +/- 0.06 (at rest) to 1.44 +/- 0.41 micromol x g(-1) x min(-1) (P < 0.01) in the contralateral hemispheric compartment (24 mm3) and to 0.65 +/- 0.10 micromol x g(-1) x min(-1) (P < 0.03) in the ipsilateral side. Each V(TCA) value was converted to the cerebral metabolic rates of glucose oxidation (oxidative-CMR(glc)) and oxygen consumption (CMR(O2)). These rates were corrected for partial-volume based on activation maps obtained by blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI). The percent increase and the absolute value of oxidative-CMR(glc) in the activated regions are similar to values reported previously for total-CMR(glc) using the same activation paradigm. This indicates that the large majority of energy required for brain activation, in going from the resting to an activated state, is supplied by glucose oxidation. The level of activity during stimulation is relevant to awake animals because the oxidative CMR(glc) (1.05 +/- 0.28 micromol x g(-1) x min(-1); current study) is in the range of total-CMR(glc) previously reported for awake rats undergoing physiologic activation (0.7-1.4 micromol x g(-1) x min(-1)). It is concluded that oxidative glycolysis is the main source of energy for increased brain activity and a positive BOLD fMRI signal-change occurs in conjunction with a large increase in CMR(O2). PMID- 9346430 TI - In vivo uptake of [3H]nimodipine in focal cerebral ischemia: modulation by hyperglycemia. AB - Cell membrane depolarization and tissue acidosis occur rapidly in severely ischemic brain. Preischemic hyperglycemia is recognized to increase ischemic tissue acidosis and the present studies were undertaken to correlate depolarization and tissue acidosis during acute focal cerebral ischemia and hyperglycemia. We used a dual-label autoradiography method to simultaneously measure the in vivo distribution of [3H]nimodipine and [14C]DMO (5,5-dimethyl-2,4 oxazolidinedione) in brain to identify regions of ischemic depolarization and measure regional net tissue pH. Regional cerebral blood flow (CBF) was measured in separate studies. Measurements were made 30 minutes after combined middle cerebral artery and ipsilateral common carotid artery occlusion in normoglycemic and hyperglycemic rats. Tissue pH in the ischemic cortex was depressed to 6.76 +/ 0.11 in normoglycemic rats (n = 12) and 6.57 +/- 0.13 in hyperglycemic rats (n = 12), with significantly greater acidosis in the hyperglycemic group (P < 0.001). In contrast the ratio of [3H]nimodipine uptake in the ischemic cortex relative to the contralateral nonischemic cortex was significantly greater in normoglycemic (1.83 +/- 0.45) than hyperglycemic (1.40 +/- 0.50) rats (P < 0.05). Within this region of ischemic cortex CBF was 31 +/- 22 mL/100 g in normoglycemic rats (n = 8) and 33 +/- 22 mL/100 g/min in hyperglycemic rats (n = 9). Cerebral blood flow did not differ between these two groups in any region. Thus hyperglycemia reduced the extent of ischemic depolarization within the cortex during the first 30 minutes of focal cerebral ischemia. This effect may be related to the increased tissue acidosis or to other factors that may lessen calcium influx and preserve cellular energy stores in the ischemic cortex of the hyperglycemic rats. PMID- 9346429 TI - Reperfusion injury: demonstration of brain damage produced by reperfusion after transient focal ischemia in rats. AB - During reperfusion after ischemia, deleterious biochemical processes can be triggered that may antagonize the beneficial effects of reperfusion. Research into the understanding and treatment of reperfusion injury (RI) is an important objective in the new era of reperfusion therapy for stroke. To investigate RI, permanent and reversible unilateral middle cerebral artery/common carotid artery (MCA/CCA) occlusion (monitored by laser Doppler) of variable duration in Long Evans (LE) and spontaneously hypertensive (SH) rats and unilateral MCA and bilateral CCA occlusion in selected LE rats was induced. In LE rats, infarct volume after 24 hours of permanent unilateral MCA/CCA occlusion was 31.1 +/- 34.6 mm3 and was only 28% of the infarct volume after 120 to 300 minutes of reversible occlusion plus 24 hours of reperfusion, indicating that 72% of the damage of ischemia/reperfusion is produced by RI. When reversible ischemia was prolonged to 480 and 1080 minutes, infarct volume was 39.6 mm3 and 16.6 mm3, respectively, being indistinguishable from the damage produced by permanent ischemia and significantly smaller than damage after 120 to 300 minutes of ischemia. Reperfusion injury was not seen in SH rats or with bilateral CCA occlusion in LE rats, in which perfusion is reduced more profoundly. Reperfusion injury was ameliorated by the protein synthesis inhibitor cycloheximide or spin-trap agent N tert-butyl-alpha-phenylnitrone pretreatment. PMID- 9346431 TI - Regional cerebral blood flow during and after 2 hours of middle cerebral artery occlusion in the rat. AB - In this study we explored if the secondary bioenergetic failure, which occurs a few hours after recirculation, following transient middle cerebral artery occlusion (MCAO) in rats, is caused by a compromised reflow. We induced 2 hours of MCAO and measured CBF at the end of the ischemia, as well as 15 minutes, 1, 2, and 4 hours after the start of recirculation, using autoradiographic or tissue sampling 14C-iodoantipyrine techniques. After 2 hours of MCAO, the autoradiographically measured CBF in the ischemic core areas was reduced to 3 to 5% of contralateral values. The reduction in CBF was less in neighboring, penumbral areas. After recirculation, flow already normalized in core tissues after 15 minutes, and remained close to normal for the 4 hours recirculation period studied. However, in penumbral tissues, recovery CBF values were usually below normal. The results show that tissues that are heavily compromised by the 2 hour period of ischemia and are destined to incur infarction, show a "relative hyperemia" during recirculation. In fact, some areas of the previously densely ischemic tissue showed overt hyperperfusion. This finding raises the question whether the relative or absolute hyperemia reflects events that are pathogenetically important. Because drugs that clearly ameliorate the final damage incurred fail to alter the relative hyperperfusion of previously ischemic tissues, it is concluded that vascular events in the reperfusion period do not play a major role in causing the final damage. PMID- 9346432 TI - L-arginine-induced regional cerebral blood flow increase is abolished after transient focal cerebral ischemia in the rat. AB - We investigated the L-arginine-induced, regional cerebral blood flow (rCBF) enhancement after different durations of transient focal cerebral ischemia in the rat to determine if L-arginine increases rCBF after transient focal cerebral ischemia. Focal ischemia (5 minutes and 20 minutes) followed by 90 minutes of reperfusion was induced in a normotensive rat suture-model. Regional cerebral blood flow in both hemispheres was measured by laser-Doppler-flowmetry. Reactivity of rCBF to L-arginine (300 mg/kg) was measured 45 minutes after reperfusion, and hypercapnia 90 minutes after reperfusion. The effect of D arginine and pretreatment with the nitric oxide (NO) synthase inhibitor N(omega) nitro-L-arginine (L-NA) (10 mg/kg) was examined in additional groups. Hypercapnia and L-arginine increased rCBF in sham operated controls and on the nonischemic hemispheres. D-arginine did not. Twenty-minute long ischemia significantly reduced the response to L-arginine (control side: 115 +/- 5.9%; ischemic side: 107 +/- 6.1%, n = 7) and hypercapnia, 5 minutes of ischemia did not. N(omega) nitro-L-arginine pretreatment partly restored the L-arginine-induced rCBF increase. Thus, rCBF increase caused by L-arginine in the reperfusion period was unaffected by 5 minutes of ischemia, but reduced by 20 minutes of ischemia. The restoration after pretreatment with L-NA may be caused by attenuated production of cytotoxic substances, e.g., NO and related compounds. PMID- 9346433 TI - A mouse model of embolic focal cerebral ischemia. AB - We developed a mouse model of embolic focal cerebral ischemia, in which a fibrin rich clot was placed at the origin of the middle cerebral artery (MCA) in C57BL/6J mice (n = 31) and B6C3 mice (n = 10). An additional three non-embolized C57BL/6J mice were used as a control. Embolus induction, cerebral vascular perfusion deficit, and consequent ischemic cell damage were confirmed by histopathology, immunohistochemistry, laser confocal microscopy, and regional cerebral blood flow (rCBF) measurements. Reduction in rCBF and cerebral infarct were not detected in the control animals. An embolus was found in all C57BL/6J and B6C3 mice at 24 hours after injection of a clot. Regional CBF in the ipsilateral parietal cortex decreased to 23% (P < 0.05) and 17% (P < 0.05) of preembolization levels immediately and persisted for at least 1 hour in C57BL/6J mice (n = 6) and in B6C3 mice (n = 3), respectively. A significant decrease of rCBF was accompanied by a corresponding reduction of plasma perfusion in the ipsilateral MCA territory. Neurons exhibited marked reduction in microtubule associated protein-2 immunostaining coincident with the area of perfusion deficit. The percent infarct volume was 30.3% +/- 13.4% for C57BL/6J mice (n = 17), and 38.3% +/- 15.3% for B6C3 mice (n = 7) at 24 hours after embolization. This model of embolic ischemia is relevant to thromboembolic stroke in humans and may be useful to investigate embolic cerebral ischemia in the genetically altered mouse and for evaluation of antiembolic therapies. PMID- 9346434 TI - Role of nitric oxide in regulation of brain stem circulation during hypotension. AB - We tested the hypothesis that nitric oxide (NO) plays a role in CBF autoregulation in the brain stem during hypotension. In anesthetized rats, local CBF to the brain stem was determined with laser-Doppler flowmetry, and diameters of the basilar artery and its branches were measured through an open cranial window during stepwise hemorrhagic hypotension. During topical application of 10( 5) mol/L and 10(-4) mol/L N(omega)-nitro-L-arginine (L-NNA), a nonselective inhibitor of nitric oxide synthase (NOS), CBF started to decrease at higher steps of mean arterial blood pressure in proportion to the concentration of L-NNA in stepwise hypotension (45 to 60 mm Hg in the 10(-5) mol/L and 60 to 75 mm Hg in the 10(-4) mol/L L-NNA group versus 30 to 45 mm Hg in the control group). Dilator response of the basilar artery to severe hypotension was significantly attenuated by topical application of L-NNA (maximum dilatation at 30 mm Hg: 16 +/- 8% in the 10(-5) mol/L and 12 +/- 5% in the 10(-4) mol/L L-NNA group versus 34 +/- 4% in the control group), but that of the branches was similar between the control and L-NNA groups. Topical application of 10(-5) mol/L 7-nitro indazole, a selective inhibitor of neuronal NOS, did not affect changes in CBF or vessel diameter through the entire pressure range. Thus, endothelial but not neuronal NO seems to take part in the regulation of CBF to the the brain stem during hypotension around the lower limits of CBF autoregulation. The role of NO in mediating dilatation in response to hypotension appears to be greater in large arteries than in small ones. PMID- 9346435 TI - Platelet-derived growth factor-BB, but not -AA, prevents delayed neuronal death after forebrain ischemia in rats. AB - Our previous studies demonstrated coordinate expression of platelet-derived growth factor (PDGF) -B chain and beta-receptor in neurons at risk in the rat brain with focal ischemia. To clarify a role of the -B chain in the brain further, we examined whether PDGF-A or -B chain protects CA1 pyramidal neurons from delayed neuronal death after forebrain ischemia in rats. Pretreatment with PDGF-BB, but not -AA, at 120 ng/d for 2 days until forebrain ischemia was performed markedly ameliorated delayed neuronal death in CA1 pyramidal neurons on day 7 after ischemia. This neuroprotective effect of PDGF-BB was dose-dependent, and pretreatment with PDGF-BB at 240 ng/d showed almost complete inhibition of delayed neuronal death. In contrast, posttreatment with PDGF-BB at 120 ng/d starting 20 minutes after ischemia demonstrated no significant neuroprotective effect. The current study established marked neuroprotective actions of PDGF-BB in ischemic neuronal damage. PMID- 9346436 TI - Global forebrain ischemia results in differential cellular expression of interleukin-1beta (IL-1beta) and its receptor at mRNA and protein level. AB - The mRNA expression of the proinflammatory cytokine interleukin-1beta (IL-1beta) has been shown to be induced in neural elements during ischemia. It is not clear which cells generate the IL-1beta mRNA and eventually synthesize IL-1 protein and which cells respond to this signaling by producing IL-1 receptors during ischemia. To clarify this question, rats were subjected to global ischemia by bilateral carotid occlusion and hypotension for 20 minutes, followed by reperfusion for 2 hours (n = 7), 8 hours (n = 7), or 24 hours (n = 7). Cryostat sections were hybridized using antisense oligonucleotide probes (30 dimer). Multiple cell markers were used in immunohistochemical staining to identify the cells expressing IL-1beta and IL-1R protein. The sham animals (n = 5) showed no or only a weak expression of IL-1R or IL-1beta mRNA. The number of IL-1beta mRNA expressing cells was significantly increased by 2 hours of reperfusion in several brain areas including cortex (12-fold compared with sham) and caudate-putamen (14 fold), and was maximally increased in most hippocampal regions by 8 hours of reperfusion (mean +/- SD of positive cells/field versus sham equivalent being 37.9 +/- 12.3 versus 4.0 +/- 3.3; 30.6 +/- 9.0 versus 3.1 +/- 2.3; 41.3 +/- 17.5 versus 2.9 +/- 1.9; in CA1; CA2; CA3/CA4 regions of the hippocampus, respectively). IL-1beta mRNA signal was also intensified in the white matter areas. Changes in IL-1R mRNA were seen in the hippocampus (after 2 hours CA1: 16 fold; CA2: 17-fold; DG: 24-fold increase; and CA3/CA4: 10-fold increase after 8 hours), and the expression was prolonged especially in CA1 and CA2 regions up to 24 hours of reperfusion. The major cellular source of IL-1beta protein was glia (astrocytes, oligodendrocytes, microglia, and scattered perivascular macrophages/monocytes), while neurons and sporadic microvascular endothelia showed IL-1R immunoreactivity. The data suggest that neurons in discrete areas vulnerable for selective neuronal death, and possibly the vascular endothelium, are target cells for ischemia-induced glial IL-1beta production. PMID- 9346437 TI - A possible mechanism for the aglycemia-induced depression of glutamatergic excitation in the striatum. AB - We have studied the possible mechanisms underlying the decrease of excitatory transmission induced by glucose deprivation by using electrophysiological recordings in corticostriatal slices. Extracellular field potentials were recorded in the striatum after cortical stimulation; these potentials were progressively reduced by glucose deprivation. The reduction started 5 minutes after the onset of aglycemia. The field potential was fully suppressed after 40 minutes of glucose deprivation. After the washout of the aglycemic solution only a partial recovery was observed. Aglycemia also induced a delayed inward current during single-microelectrode voltage-clamp recordings from spiny neurons. This inward current was coupled with an increased membrane conductance. The A1 adenosine receptor antagonists, 8-cyclopentyl-1,3-dimethylxanthine (CPT, 1 micromol/L) and 1,3-dipropyl-8-cyclopentylxanthine (CPX, 300 nmol/L), significantly reduced the aglycemia-induced decrease of field potential amplitude. Moreover, in the presence of CPT and CPX, a full recovery of the field potential amplitude after the interruption of the aglycemic solution was observed. Conversely, these antagonists affected neither the inward current nor the underlying conductance increase produced by glucose deprivation. The ATP sensitive potassium channel blockers glibenclamide (10 micromol/L) and glipizide (100 nmol/L) had no effect on the aglycemia-induced decrease of the field potential amplitude. We suggest that endogenous adenosine, but not ATP-dependent potassium channels, plays a significant role in the aglycemia-induced depression of excitatory transmission at corticostriatal synapses probably through a presynaptic mechanism. Moreover, adenosine is not involved in the postsynaptic changes induced by glucose deprivation in spiny striatal neurons. PMID- 9346438 TI - Frequency dependence of cerebrovascular impedance in preterm neonates: a different view on critical closing pressure. AB - The nonproportional relationship between instantaneous arterial blood pressure (BP) and cerebral blood flow velocity (CBFv) is well explained by the concept of critical closing pressure (CCP). We aimed to determine the frequency response of the neonatal cerebrovascular system, and to establish the exact mathematical relationship between cerebrovascular impedance and CCP under physiologic conditions. In 10 preterm neonates (gestational age, 25-32 weeks; birth weight, 685-1,730 g; age 1-7 days) we Doppler-traced CBFv of the internal carotid artery. Blood pressure was traced simultaneously. Critical closing pressure was graphically determined. Cerebrovascular impedance was calculated as the square root of the ratio of the corresponding peaks in the power spectra of BP and CBFv at zero frequency, and at heart rate (H) and harmonics (xH). Uniformly, the impedance between H and 3H (2 to 6 Hz) was reduced about fivefold, compared with the impedance at zero frequency. The cerebrovascular system behaves like a high pass filter, leading to a reduction of the DC (direct current) component of CBFv (analogous to current) relative to that of the driving force BP (analogous to voltage). The frequency response of cerebrovascular impedance reflects the ratio of CCP and DC BP. A mathematical derivation of this relationship is given matching the observed results. Thus, both the CCP and the impedance approach are valid. PMID- 9346439 TI - Delayed triphenyltetrazolium chloride staining remains useful for evaluating cerebral infarct volume in a rat stroke model. AB - Sixteen of 24 Sprague-Dawley rats with permanent middle cerebral artery occlusion for 24 hours were subjected to immediate or 8-hour delayed 2,3,5 triphenyltetrazolium chloride (TTC) staining (n = 8 at each time point); the other 8 animals were subjected to immediate or 8-hour delayed measurement of succinate dehydrogenase activity (n = 4 at each time point). The TTC staining was of good quality good in all animals, and the infarcted region could be distinguished easily from normal tissue. There was no significant difference in corrected infarct volume between the two groups (263.8 +/- 43.1 versus 264.4 +/- 54.8 mm3 [mean +/- standard deviation]). The activity of succinate dehydrogenase was not significantly different when normal or infarcted tissue was measured immediately after death or with an 8 hour delay, although less activity was detected at both time points in the infarcted tissue. These results demonstrate that an 8-hour delay of TTC staining is reliable for evaluating brain infarct volume in a rat stroke model and this probably is attributable to the slow deterioration of mitochondrial enzyme activity in nonischemic brain over this time period. PMID- 9346454 TI - Effect of two aspirin pretreatment regimens on niacin-induced cutaneous reactions. AB - OBJECTIVE: To compare the effects of pretreatment with two aspirin regimens and placebo on niacin-induced cutaneous reactions. DESIGN: Randomized, double-blind, placebo-controlled, crossover study. SETTING: Internal medicine clinic in an academic health center. PARTICIPANTS: Forty-two healthy subjects (22 males and 20 females) between the ages of 35 and 65 (mean age 44.2 years) were recruited and completed the study. Subjects received aspirin 325 mg, aspirin 650 mg, and placebo for 4 consecutive days, and on the fourth day also ingested 500 mg of immediate-release niacin 30 minutes after taking aspirin or placebo. They reported the intensity of flushing, headache, pruritus, tingling, and warmth on a 10-cm visual analogue scale. Reactions were evaluated at time 0 (before the niacin dose), and at 15, 30, 60, and 120 minutes following the niacin dose. Cutaneous reactions were compared at each evaluation time and scored by two other methods. The peak intensity was the highest score recorded at any of the four evaluation times after niacin administration. An intensity-time factor was calculated by totaling the scores of each of the four evaluation times. MEASUREMENT AND MAIN RESULTS: The symptom scores for flushing, itching, tingling, and warmth were all significantly reduced by both aspirin regimens (p < .05 in all cases), although there were no significant differences between the 325-mg and 650-mg doses. The results were similar for each scoring method. CONCLUSIONS: An aspirin regimen of 325 mg is effective in suppressing niacin-induced cutaneous reactions. Increasing the dose to 650 mg does not provide additional benefit. PMID- 9346455 TI - Effects of a self-administered previsit questionnaire to enhance awareness of patients' concerns in primary care. AB - OBJECTIVE: To determine if a self-administered previsit questionnaire designed to increase awareness of patients' concerns alters the visit duration, content of the discussion, and patient and physician satisfaction. DESIGN: A balanced, two arm trial in which physicians were randomized. SETTING: Two primary-care clinics affiliated with a university hospital. PATIENTS/PARTICIPANTS: Ten physicians and 201 continuity-care patients. INTERVENTIONS: In intervention visits, patients completed a previsit questionnaire asking about the desire for medical information, psychosocial assistance, therapeutic listening, general health advice, and biomedical treatment. Physicians reviewed questionnaires with patients during the visit. MEASUREMENTS AND MAIN RESULTS: We used audiotapes of encounters to quantify the duration of the encounter and measured the number and type of diagnoses discussed in the visit, and patient and physician satisfaction with the encounter. Intervention visits were 34% longer (increase of 6.8 minutes; 95% confidence interval [CI] 0.4, 13.2) than control visits with most of the additional time spent in discussion of biomedical diagnoses (3.35 minutes; 95% CI 0.00, 6.72) and in the performance of the physical examination (2.7 minutes; 95% CI 0.5, 4.9). The number of diagnoses discussed per visit was 30% higher in intervention visits (increase of 1.7 diagnoses per visit; 95% CI 0.3, 3.2), but patients' satisfaction with these visits tended to be lower. CONCLUSIONS: Using a previsit questionnaire to increase awareness of the patients' concerns may entail a trade-off between conflicting goals: trying to respond to patient concerns while not significantly increasing the cost per visit. A future challenge is to develop and refine techniques with sufficient efficacy to justify the expense of implementing the intervention and the longer visit needed to respond adequately to patients' concerns. PMID- 9346456 TI - What is a screening test? Misclassification bias in observational studies of screening for cancer. AB - OBJECTIVE: To demonstrate the importance of accurately identifying clinical distinctions of subjects in observational studies of screening. DESIGN: Simulated case-control studies. SETTING: The West Haven Veterans Affairs Medical Center. PATIENTS: Fifty-two men diagnosed with prostate cancer in 1988 or 1989 had 252 digital rectal examinations (DREs) in the preceding 5 years. A classification scheme used patient symptoms and the results of prior DREs to assign the last DRE before the diagnosis of cancer to one of the following categories: definite screening, likely screening, probable screening, not screening, or other and unknown. Sixty-five percent of the DREs were classified as definite or likely screening, and another 15% were classified as probable screening. MAIN RESULTS: Changing the definition of a screening DRE from one including to one excluding probable DREs lowered the frequency of screening in case subjects more than it did in case controls, and thus lowered the odds ratio (OR), making screening appear to be more protective. Even when DRE was not protective, the ORs for the effectiveness of screening with the more restrictive definition ranged from 0.21 to 0.83 in 36 simulated case-control studies that differed according to the frequency of screening, the prevalence of cancer in case controls, and the extent of misclassification error. CONCLUSIONS: If clinical distinctions in the performance of screening tests are not classified appropriately, observational studies will misrepresent the proportion of subjects exposed to screening interventions and produce biased results. PMID- 9346457 TI - Perceived adequacy of tangible social support and health outcomes in patients with coronary artery disease. AB - OBJECTIVE: Health outcomes of patients with chronic disease might be influenced by assistance from others in performing daily activities. We examined whether perceived adequacy of such tangible support was associated with prognosis in a cohort of patients with coronary artery disease. DESIGN: Longitudinal cohort study. SETTING/PARTICIPANTS: In spring 1993, a cohort of 1,468 patients with chronic artery disease was identified using claims data. The cohort consisted of all surviving residents of Manitoba, Canada, who had been hospitalized for acute myocardial infarction from 1991 to 1992: 820 patients completed the initial survey, and 734 completed a follow-up survey approximately 1 year later. MEASUREMENTS AND MAIN RESULTS: Adequacy of tangible support was assessed by asking if respondents needed help at home because of health problems, and whether these needs were met. We examined the association between perceived adequacy of tangible support and health outcomes at 1 year (mortality, physical function). Of 820 participants, 74% perceived no need for help, 13% had sufficient help, 9% needed more help, and 5% needed much more help; 31 patients died during follow up. After adjustment for age and initial health status, odds ratios (95% confidence interval) for death were: sufficient help 1.8 (0.61, 5.8); need more help 3.2 (1.1, 9.4); and need much more help 6.5 (2.0, 21.6) compared with respondents with no perceived need. Decline in physical function was also linearly related to perceiving less-adequate tangible support. Sensitivity analyses indicated it is highly improbable that results were due to selection bias. CONCLUSIONS: Perceived lack of needed assistance was related to mortality and to decline in physical functioning. Adequacy of tangible support was an important prognostic factor for these patients with coronary artery disease and may be a determinant of health outcomes. PMID- 9346458 TI - Primary care physicians' use of lumbar spine imaging tests: effects of guidelines and practice pattern feedback. AB - OBJECTIVE: To reduce variability in primary care physicians' use of procedures for imaging the lumbar spine. DESIGN: Controlled intervention using clinical practice guideline and practice pattern feedback. STUDY SAMPLE: Sixty-seven internists and 28 family practitioners in a large, group-model HMO. MEASUREMENTS AND MAIN RESULTS: Intervention group physicians received the clinical practice guideline for low back pain, followed after 4 months by three bimonthly feedback reports on their current use rates for lumber spine x-rays and computed tomography and magnetic resonance imaging scans of the lumbar spine. Control group physicians received neither the guideline nor the feedback reports. Automated radiology utilization data were used to compare intervention and control group physicians' changes in use rates and variability in use rates over the course of the study period. Neither the guideline alone nor the guideline plus feedback was associated with a significant decrease in use rates or in the variability in use rates for the lumbar spine imaging procedures under study. CONCLUSIONS: Clinical practice guidelines and practice pattern feedback fail to achieve their goals when features of the practice setting and patient expectations and behavior are not identified and addressed. PMID- 9346459 TI - Translating statistics for use in the clinic. PMID- 9346460 TI - Primary care for the developmentally disabled adult. PMID- 9346461 TI - Effects of an educational intervention on residents' knowledge and attitudes toward interactions with pharmaceutical representatives. AB - To assess primary care resident and faculty knowledge and attitudes concerning interactions between physicians and pharmaceutical representatives (PRs) and to measure changes in residents' knowledge and attitudes after an educational intervention, we conducted preintervention and postintervention surveys with a causal-comparative group in a university-based primary care residency program. All primary care internal medicine and internal medicine-pediatrics residents and faculty were given the voluntary survey. In general, residents and faculty demonstrated similar responses for the preintervention survey. Differences between faculty and resident opinions were seen in two areas. Faculty were more likely than residents to believe that PRs sometimes use unethical marketing practices (p < .05) and that the amount of contact with PRs in the outpatient clinic is excessive (p < .01). The postintervention survey of residents demonstrated significant differences between the control and intervention groups for three attitude scales. After the intervention, residents showed an increased belief that PRs may use unethical marketing practices (p < .01), that marketing gifts with no patient benefit may be inappropriate (p = .05), and that other physicians' prescribing patterns could be negatively influenced through the acceptance of gifts (p < .05). A brief educational intervention can change resident attitudes concerning physician interactions with PRs. PMID- 9346462 TI - Probable gastrointestinal toxicity of Kombucha tea: is this beverage healthy or harmful? AB - Kombucha tea is a health beverage made by incubating the Kombucha "mushroom" in tea and sugar. Although therapeutic benefits have been attributed to the drink, neither its beneficial effects nor adverse side effects have been reported widely in the scientific literature. Side effects probably related to consumption of Kombucha tea are reported in four patients. Two presented with symptoms of allergic reaction, the third with jaundice, and the fourth with nausea, vomiting, and head and neck pain. In all four, use of Kombucha tea in proximity to onset of symptoms and symptom resolution on cessation of tea drinking suggest a probable etiologic association. PMID- 9346463 TI - Hyperkalemia in the elderly: drugs exacerbate impaired potassium homeostasis. AB - OBJECTIVE: To review the pathophysiology underlying the predisposition to hyperkalemia in the elderly; the medications that disrupt potassium balance and promote the development of hyperkalemia in the elderly; the prevention of hyperkalemia in elderly patients treated with potassium-altering medications; and the appropriate management of hyperkalemia when it develops. METHODS AND MAIN RESULTS: A MEDLINE search of the literature (1966-1996) using the terms hyperkalemia, drugs, elderly, and treatment was conducted and pertinent review articles, textbooks, and personal files were consulted. Elderly subjects appear to be predisposed to the development of hyperkalemia on the basis of both innate disturbances in potassium homeostasis and comorbid disease processes that impair potassium handling. Hyperkalemia in the elderly is most often precipitated by medications that impair cellular uptake or renal disposal of potassium. This electrolyte disorder is best prevented by recognition of at-risk physiology in the aged, avoidance of therapy with certain high-risk medications, and monitoring of plasma potassium concentration and renal function at intervals appropriate for the medication prescribed. Management of hyperkalemia entails identification of the clinical manifestations of severe hyperkalemia, stabilization of cardiac tissue, promotion of cellular potassium uptake, and ultimately removal of potassium from the body. CONCLUSIONS: Geriatric patients should be considered at risk of developing hyperkalemia, especially when they are prescribed certain medications. Potassium levels should be monitored at appropriate intervals when these patients are treated with potassium-altering medications. Appropriate management of hyperkalemia in the elderly can avoid life-threatening neuromuscular and cardiac complications. PMID- 9346464 TI - Is there an easier way to determine whether early detection of prostate cancer reduces mortality? PMID- 9346465 TI - Questioning decision analysis for treatment of clinically localized prostate cancer. PMID- 9346466 TI - Ornithine transcarbamylase deficiency: pathogenesis of the cerebral disorder and new prospects for therapy. AB - Ornithine Transcarbamylase (OTC) is a key urea cycle enzyme. Congenital OTC deficiencies in humans result in hyperammonemia and a spectrum of neurological symptoms including hypotonia, seizures and mental retardation. Neuropathologic evaluation reveals cerebral atrophy, ventricular enlargement and Alzheimer type II astrocytosis. Using an animal model of congenital OTC deficiency, the sparse fur (spf) mouse, recent studies have revealed significant alterations of cholinergic, serotoninergic and glutamatergic neurotransmitter systems. Possible pathophysiologic mechanisms responsible for neuronal cell loss in OTC deficiency include a deficit in cerebral energy metabolism, and glutamate excitotoxicity. Therapy continues to rely on alternative substrate administration including sodium benzoate and sodium phenylacetate. Experimental evidence suggests that acetyl-L-carnitine and glutamate (NMDA) receptor antagonists could be potentially useful therapeutic agents. Liver transplantation is effective in many patients and recent experimental studies using adenoviral vectors suggest that gene therapy may ultimately be useful in the treatment of congenital OTC deficiency. PMID- 9346467 TI - Wernicke's encephalopathy: an excitotoxicity hypothesis. AB - Thiamine deficiency is a recognized cause of Wernicke's encephalopathy (WE), a condition in which small necrotic lesions are found in close proximity to the third and fourth ventricles and the Sylvian aqueduct. Although the neuropathology of WE is well-established, the pathogenic mechanisms that determine the formation and distribution of brain lesions identified with this illness are not understood. It is proposed here that glutamate neurotoxicity causes the brain lesions in WE. Glutamic acid decarboxylase (GAD), an enzyme mainly confined to the central nervous system, protects most regions of the brain from glutamate that accumulates when the activity of alpha-ketoglutarate dehydrogenase, a thiamine-dependent enzyme complex, is reduced. During severe thiamine deficiency, glutamate accumulates in GAD-free peripheral tissues and reaches a concentration in blood at which it passes through circumventricular organs into the cerebral ventricles or contiguous brain and finally diffuses into the extracellular space of proximate diencephalic and brain stem tissues. Extracellular glutamate eventually reaches neurotoxic levels in those tissues and causes the characteristic lesions of WE. PMID- 9346468 TI - Potassium-evoked neuronal release of serotonin in experimental chronic portal systemic encephalopathy. AB - Portal-systemic encephalopathy (PSE) is associated with an increased brain tissue turnover of serotonin (5-HT). Despite increased 5-HT metabolism, brain 5-HT release in rats with a portacaval shunt (PCS) seems to be unaltered. Although this may indicate that the overall 5-HT output is unaltered in PSE, it is also possible that the 5-HT release pattern might be altered in some way. In the present study, the potassium-evoked frontal neocortical release of 5-HT was studied in experimental chronic PSE. KCI (60 mM) produced marked increases in the 5-HT output compared with basal values both in PCS and sham rats. Simultaneously, the KCI challenge resulted in significant elevations in the 5-HT release of PCS compared with sham. In Ca2+-free medium, the difference between PCS and sham rats in the KCl-evoked release of 5-HT was abolished. In the presence of TTX (1 mM), both groups displayed increased extracellular 5-HT levels. Again, a difference with higher amplitude of the 5-HT release in PCS compared with sham was evident. It is concluded that in experimental chronic PSE an augmented neocortical 5-HT release compared with the normal in vivo situation is available. The possible mechanism(s) responsible for the difference in neocortical 5-HT output between PCS and sham-operated rats in response to the KCl-challenge is discussed. PMID- 9346469 TI - Effect of heat shock on neuronal cultures: importance of protein synthesis and HSP72 induction for induced tolerance and survival. AB - In this study the effects of 30 min heat-shock, ranging from 42 degrees C to 46 degrees C, on survival, protein synthesis and HSP72 expression were investigated in primary rat neuronal cultures. Heat-shock of 44 degrees C resulted in a complete, but transient inhibition of protein synthesis which recovered within 24 h. 46 degrees C heat-shock resulted in an irreversible inhibition of protein synthesis and complete neuronal loss within 24 h. Cycloheximide treatment of neuronal cultures resulted in aggravation of neuronal cell damage after heat shock of 44 degrees C, indicating that the capacity for recovery of the overall protein synthesis is an important survival factor. In addition, the reduction of neuronal cell damage mediated by heat conditioning was abolished by cycloheximide treatment, indicating that the function of new proteins is important for induced thermotolerance. Induction of the strictly inducible member of the heat-shock protein 70kDa family, HSP72, was found in those few astrocytes which were contaminating the neuronal cell cultures, but not in neurons. These results indicate that newly synthesised proteins other than HSP72 are likely to mediate neuronal protection following heat shock in our experiments. These findings raise the possibility that induced tolerance may not necessarily be mediated by HSP72. PMID- 9346470 TI - Penetration of glutamate into brain of 7-day-old rats. AB - The permeability of the blood-brain barrier to glutamate was measured by quantitative autoradiography in brains of 7-day-old rats (average plasma glutamate 114 microM) and rats injected subcutaneously with glutamate (average plasma glutamate 2,670 microM). Measurements of glutamate permeability were initiated by the injection of [14C]glutamate into the inferior vena cava and the 7-day-old rats sacrificed at 1 minute to avoid the accumulation of [14C]glutamate metabolites in plasma. Glutamate entered the brain at a slow rate, with an average permeability-surface area product of 12 microl x min(-1) x g(-1), except in those areas known to have fenestrated capillaries. Thus, glutamate readily entered and accumulated in circumventricular organs where the radioactivity was localized. Although three areas with a blood-brain barrier, the cerebral cortex, the hypothalamus and the midbrain, of 7-day-old rats had permeabilities similar to adult rats, the other areas of the brain with a blood-brain barrier had a permeability about 1.5-1.9 times that of adult rats. The greater permeability of the brain of 7-day-old rats may reflect the degree of immaturity of the blood brain barrier. PMID- 9346471 TI - p-Chloroamphetamine- and d-fenfluramine-induced brain serotonin release in experimental portal-systemic encephalopathy. AB - Portal-systemic encephalopathy (PSE) is associated with increased brain turnover of serotonin (5-HT) in vivo but the brain 5-HT output seems to be unaltered. Recent results suggest, however, that an augmented neocortical 5-HT release in experimental chronic PSE may prevail under certain conditions. In the present study, neocortical extracellular 5-HT and 5-hydroxyindoleacetic-3-acid (5-HIAA) levels were measured in portacaval shunted (PCS) rats and sham-operated controls following local administration of p-chloroamphetamine (pCA) and d-fenfluramine (dFEN), two specific 5-HT releasing agents. The basal neocortical extracellular 5 HT concentrations were unaltered and the 5-HIAA levels were elevated in experimental PSE, supporting an unchanged brain 5-HT output despite elevated brain 5-HT metabolism. Perfusion with pCA or dFEN (5 microM; one 20-min pulse) produced marked increases in brain 5-HT release both in PCS and sham-operated rats compared with corresponding basal values. While no difference in the 5-HT response to dFEN administration was seen between sham (5-HT levels increased by 330%) and PCS (500%) rats, a clear difference (p<0.05) in the brain 5-HT output was observed between the two experimental groups following pCA perfusion (sham, 1100% versus PCS, 1470%). These results support our previous contention of an enhanced neocortical 5-HT output in experimental chronic PSE under certain pharmacological conditions. PMID- 9346472 TI - Effects of combined glutamate and platelet-activating factor inhibition on the outcome of focal cerebral ischaemia - an initial screening study. AB - Since both glutamate excitotoxicity and inflammatory responses have been implicated in ischaemic neuronal death, we questioned whether joint inhibition of both processes would be more neuroprotective than either on its own. Therefore we assessed the effects of combined inhibition of both glutamate release (with a use dependant sodium channel blocker, 619C89) and inflammatory processes (with a platelet-activating factor (PAF) receptor antagonist, BB-823) on the degree of motor deficit and the extent of cerebral (cortical and sub-cortical grey matter) infarction produced by middle cerebral artery occlusion (MCAO) in the rat, and compared results to appropriate single agent, vehicle and positive controls. The combination of both agents produced the greatest reduction in motor deficit, but the effect was only significant (p<0.05) acutely (4 to 6 hours post-MCAO). The extent of cortical infarction at 24 hours post-MCAO was significantly reduced in all experimental groups compared to vehicle-controls (p<0.05) and the greatest reduction occurred in the combination group (55%), though it was not significantly better than either of the single agent groups. Similarly the greatest reduction in sub-cortical infarction was in the combination group, but this was also not significantly better than the single agents. The results of this novel combination of pharmacological interventions suggest that inhibition of both glutamate excitotoxicity and inflammatory responses afforded an overall enhanced, if modest, neuroprotective effect, compared to inhibition of either process alone. The possible mechanisms involved are discussed, but warrant further clarification before therapeutic strategies are developed. PMID- 9346474 TI - The impact of aerosols on solar ultraviolet radiation and photochemical smog. AB - Photochemical smog, or ground-level ozone, has been the most recalcitrant of air pollution problems, but reductions in emissions of sulfur and hydrocarbons may yield unanticipated benefits in air quality. While sulfate and some organic aerosol particles scatter solar radiation back into space and can cool Earth's surface, they also change the actinic flux of ultraviolet (UV) radiation. Observations and numerical models show that UV-scattering particles in the boundary layer accelerate photochemical reactions and smog production, but UV absorbing aerosols such as mineral dust and soot inhibit smog production. Results could have major implications for the control of air pollution. PMID- 9346478 TI - A porous silicon-based optical interferometric biosensor. AB - A biosensor has been developed based on induced wavelength shifts in the Fabry Perot fringes in the visible-light reflection spectrum of appropriately derivatized thin films of porous silicon semiconductors. Binding of molecules induced changes in the refractive index of the porous silicon. The validity and sensitivity of the system are demonstrated for small organic molecules (biotin and digoxigenin), 16-nucleotide DNA oligomers, and proteins (streptavidin and antibodies) at pico- and femtomolar analyte concentrations. The sensor is also highly effective for detecting single and multilayered molecular assemblies. PMID- 9346480 TI - Adiabatic electron transfer: comparison of modified theory with experiment. AB - The radical cations of properly designed bishydrazines allow comparison of observed and calculated electron transfer rate constants. These compounds have rate constants small enough to be measured by dynamic electron spin resonance spectroscopy and show charge transfer bands corresponding to vertical excitation from the energy well for the charge occurring upon one hydrazine unit to that for the electron-transferred species. Analysis of the data for all six compounds studied indicates that the shape of the adiabatic surface on which electron transfer occurs can be obtained from the charge transfer band accurately enough to successfully predict the electron transfer rate constant and that explicit tunneling corrections are not required for these compounds. PMID- 9346481 TI - Structure of the carboxyl-terminal dimerization domain of the HIV-1 capsid protein. AB - The carboxyl-terminal domain, residues 146 to 231, of the human immunodeficiency virus-1 (HIV-1) capsid protein [CA(146-231)] is required for capsid dimerization and viral assembly. This domain contains a stretch of 20 residues, called the major homology region (MHR), which is conserved across retroviruses and is essential for viral assembly, maturation, and infectivity. The crystal structures of CA(146-231) and CA(151-231) reveal that the globular domain is composed of four helices and an extended amino-terminal strand. CA(146-231) dimerizes through parallel packing of helix 2 across a dyad. The MHR is distinct from the dimer interface and instead forms an intricate hydrogen-bonding network that interconnects strand 1 and helices 1 and 2. Alignment of the CA(146-231) dimer with the crystal structure of the capsid amino-terminal domain provides a model for the intact protein and extends models for assembly of the central conical core of HIV-1. PMID- 9346482 TI - Metal ion chaperone function of the soluble Cu(I) receptor Atx1. AB - Reactive and potentially toxic cofactors such as copper ions are imported into eukaryotic cells and incorporated into target proteins by unknown mechanisms. Atx1, a prototypical copper chaperone protein from yeast, has now been shown to act as a soluble cytoplasmic copper(I) receptor that can adopt either a two- or three-coordinate metal center in the active site. Atx1 also associated directly with the Atx1-like cytosolic domains of Ccc2, a vesicular protein defined in genetic studies as a member of the copper-trafficking pathway. The unusual structure and dynamics of Atx1 suggest a copper exchange function for this protein and related domains in the Menkes and Wilson disease proteins. PMID- 9346483 TI - Measurement of the force-velocity relation for growing microtubules. AB - Forces generated by protein polymerization are important for various forms of cellular motility. Assembling microtubules, for instance, are believed to exert pushing forces on chromosomes during mitosis. The force that a single microtubule can generate was measured by attaching microtubules to a substrate at one end and causing them to push against a microfabricated rigid barrier at the other end. The subsequent buckling of the microtubules was analyzed to determine both the force on each microtubule end and the growth velocity. The growth velocity decreased from 1.2 micrometers per minute at zero force to 0.2 micrometer per minute at forces of 3 to 4 piconewtons. The force-velocity relation fits well to a decaying exponential, in agreement with theoretical models, but the rate of decay is faster than predicted. PMID- 9346484 TI - IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation. AB - Activation of the transcription factor nuclear factor kappa B (NF-kappaB) is controlled by sequential phosphorylation, ubiquitination, and degradation of its inhibitory subunit IkappaB. A large multiprotein complex, the IkappaB kinase (IKK) signalsome, was purified from HeLa cells and found to contain a cytokine inducible IkappaB kinase activity that phosphorylates IkappaB-alpha and IkappaB beta. Two components of the IKK signalsome, IKK-1 and IKK-2, were identified as closely related protein serine kinases containing leucine zipper and helix-loop helix protein interaction motifs. Mutant versions of IKK-2 had pronounced effects on RelA nuclear translocation and NF-kappaB-dependent reporter activity, consistent with a critical role for the IKK kinases in the NF-kappaB signaling pathway. PMID- 9346485 TI - IkappaB kinase-beta: NF-kappaB activation and complex formation with IkappaB kinase-alpha and NIK. AB - Activation of the transcription factor nuclear factor kappa B (NF-kappaB) by inflammatory cytokines requires the successive action of NF-kappaB-inducing kinase (NIK) and IkappaB kinase-alpha (IKK-alpha). A widely expressed protein kinase was identified that is 52 percent identical to IKK-alpha. IkappaB kinase beta (IKK-beta) activated NF-kappaB when overexpressed and phosphorylated serine residues 32 and 36 of IkappaB-alpha and serines 19 and 23 of IkappaB-beta. The activity of IKK-beta was stimulated by tumor necrosis factor and interleukin-1 treatment. IKK-alpha and IKK-beta formed heterodimers that interacted with NIK. Overexpression of a catalytically inactive form of IKK-beta blocked cytokine induced NF-kappaB activation. Thus, an active IkappaB kinase complex may require three distinct protein kinases. PMID- 9346487 TI - Knockout-transgenic mouse model of sickle cell disease. AB - When transgenic mice that expressed human sickle hemoglobin were mated with mice having knockout mutations of the mouse alpha- and beta-globin genes, animals were produced that synthesized only human hemoglobin in adult red blood cells. Similar to many human patients with sickle cell disease, the mice developed a severe hemolytic anemia and extensive organ pathology. Numerous sickled erythrocytes were observed in peripheral blood. Although chronically anemic, most animals survived for 2 to 9 months and were fertile. Drug and genetic therapies can now be tested in this mouse model of sickle cell disease. PMID- 9346488 TI - Transgenic knockout mice with exclusively human sickle hemoglobin and sickle cell disease. AB - To create mice expressing exclusively human sickle hemoglobin (HbS), transgenic mice expressing human alpha-, gamma-, and betaS-globin were generated and bred with knockout mice that had deletions of the murine alpha- and beta-globin genes. These sickle cell mice have the major features (irreversibly sickled red cells, anemia, multiorgan pathology) found in humans with sickle cell disease and, as such, represent a useful in vivo system to accelerate the development of improved therapies for this common genetic disease. PMID- 9346534 TI - Risk and prognostic factors of gut perforation after orthotopic liver transplantation for biliary atresia. AB - The aim of this study was to assess the risk and prognostic factors of gut perforation after orthotopic liver transplantation in children with biliary, atresia using univariate and stepwise regression analysis. Among 51 pediatric recipients who underwent transplantation because of biliary atresia after failure of portoenterostomy, 10 patients (20%) had 19 episodes of gut perforations after 14 transplantations. The median delay between transplantation and perforation was 13 days. These perforations were treated either by suture (n = 21) or ostomy (n = 11). The study of preoperative and perioperative variables showed that children with gut perforation were in surgery for a significantly longer period of time including a longer period of receiving hepatectomy and undergoing portal venous clamp. These children also needed large amounts of blood transfused during hepatectomy. After transplantation there was no difference regarding total steroid doses and early occurrence of cytomegalovirus disease between the two groups. Stepwise regression analysis identified three factors associated with the occurrence of gut perforation: duration of transplant operation, posttransplant intra-abdominal bleeding requiring reoperation, and early portal vein thrombosis. During the postoperative course, severe fungal infections were significantly more frequent in the gut perforation group. The 3-year patient survival rate was 70% in the group with gut perforation and was not different from the group without perforation (80%). This study shows that children with previous portoenterostomy carry a high risk of developing gut perforation after liver transplantation. This is especially true for those patients with the most difficult hepatectomies, which are responsible for the iatrogenic injury of the bowel. Other risk factors pointed out in this study were splanchnic congestion in case of prolonged portal venous clamp time or early portal vein thrombosis and repeated trauma of the bowel caused by reoperations. On the other hand, other well known risk factors, such as steroid therapy and viral diseases, were not involved in the occurrence of gut perforations in this study. Besides emergent surgical treatment, this type of complication requires aggressive therapy against fungal infections. PMID- 9346535 TI - Acute cellular rejection after liver transplantation: variability, morbidity, and mortality. AB - Acute cellular rejection of the allograft is a potentially serious complication after liver transplantation, yet its true incidence is unknown. We therefore investigated the frequency of acute cellular rejection reported by transplant centers and its impact on morbidity and mortality. Morbidity was defined as duration of hospitalization. Of 200 articles screened, 18 were selected for inclusion in the study database, in which there was a total of 1,437 patients who received transplants. All contained more than 20 patients and invariably used histopathology for diagnosis of acute cellular rejection. These reports included all transplant patients within a fixed period and sufficient data to determine the incidence of acute cellular rejection. Morbidity data were obtained from our previous series. The mean incidence of acute cellular rejection in all centers was 49.8% (range between centers, 24% to 80%). Two immunosuppressive cohorts were identified: high-dose cyclosporine induction (> or = 5 mg/kg/d) and low-dose cyclosporine induction (< or = 4 mg/kg/d). Acute cellular rejection was reported in 27.0% of the high-dose group and 63.6% of the low-dose group, P = .0001. Strict adherence to Snover's histological criteria for acute cellular rejection did not alter the reported mean incidence. Frequency of acute cellular rejection was 45.2% (range between centers, 24% to 80%) in 8 studies that used Snover's criteria, and 51.6% (range between centers, 37% to 80%) in 10 studies that did not. There was no correlation between mortality and incidence of acute cellular rejection in the 9 studies that reported survival (R2 = .105). Morbidity data showed that the average length of initial hospitalization after transplantation for patients with acute cellular rejection was 52.4 +/- 8.3 (range, 14 to 124) days, in contrast to 28.3 +/- 2.3 (range, 9 to 87) days for patients with no rejection. P = .0008. The total number of hospital days in the first 6 months for patients with acute cellular rejection was 55.6 +/- 8.6 (range, 14 to 124) days and with no rejection, was 37.7 +/- 3.1 (range, 9 to 99) days. P = .0232. The incidence of acute cellular rejection varies widely among transplant centers, regardless of the use of Snover's criteria. Acute cellular rejection appeared to be less frequent in programs using high-dose cyclosporine induction regimens. The presence of acute cellular rejection seemed to have no correlation with mortality but significantly increased morbidity and therefore the cost of transplantation. PMID- 9346536 TI - Intragraft cytokine gene expression in human liver allografts. AB - Cytokines are thought to play an important role in the inflammatory and immune responses of allograft rejection. We evaluated the pattern of cytokine gene expression in 36 liver biopsy specimens obtained from 20 recipients of primary orthotopic liver allografts. Specific mRNA expression was identified by a polymerase chain reaction (PCR) using oligonucleotide primers specific for human interleukin (IL)-1 beta, IL-2, IL-4, IL-6, IL-10 Interferon (IFN) gamma, tumor necrosis factor (TNF)-alpha and beta-actin. We detected IL-1 beta, IL-6 and IFN gamma cytokine message most consistently in patients with rejecting liver allografts. TNF-alpha and IL-2 were also observed in rejecting livers, but only during the early phases of the reaction. IL-4 was expressed in the majority of liver allograft biopsy specimens, regardless of the presence or absence of clinical or pathological evidence of rejection. Sequential biopsy specimens in rejecting allografts showed decreased cytokine expression after the induction of a positive response to immunosuppressive therapy. The analysis of biopsy specimens from stable liver grafts showed a predominance in the expression of IL 10. These results may reflect a differential production of inflammatory and regulatory cytokines in response to liver allograft rejection in transplant recipients. They suggest that three cytokines, IL-1 beta, IL-6 and IFN-gamma, may play an important role as markers for liver allograft rejection. Conversely, IL 10 expression was noted in patients with stable graft function. This pattern of expression may correlate with host immune responses that allow for prolonged, rejection-free survival of the graft. PMID- 9346537 TI - Liver transplantation in the presence of situs inversus totalis: application of reduced-size graft. AB - Because of the anatomical features associated with situs inversus, technical difficulties will be encountered during orthotopic liver transplantation. This report describes the case of a patient with situs inversus totalis and end-stage liver disease from biliary atresia who was treated by segmental orthotopic liver transplantation. The segmental graft was safely placed in the left subphrenic space, and a suitable orientation was obtained for anastomoses of the hilar vessels. Chronic rejection necessitated retransplantation, by the same method, 19 months later. This technique has potential advantages in coping with anatomical obstacles encountered in patients with situs inversus. PMID- 9346538 TI - Pancreatic complications after distal splenorenal shunt. AB - Pancreatic complications after the distal splenorenal shunt have not been commonly recognized. Between January 1978 and June 1993, 154 patients underwent a distal splenorenal shunt, and 11 patients (7%) developed pancreatic complications, of which 4 had pancreatitis alone, and 7 developed pancreatitis related complications. Etiology of cirrhosis, Child's classification and timing of surgery were not predictive of pancreatic complications. Eight patients (5%) were found to have chronic pancreatitis at the time of surgery, and four of these patients (50%) developed pancreatic complications following distal splenorenal shunt. Eleven early postoperative deaths in our series resulted in an overall operative mortality rate of 7%. Of these eleven patients, 6 (55%) had postoperative pancreatic complications. The operative mortality rate of patients who developed pancreatic complications (55%) after distal splenorenal shunt was significantly greater than that of patients who did not develop pancreatic complications (3%), P < .001. When compared with patients without pancreatitis, those with pancreatitis had significantly greater incidences of complete or partial portal vein thrombosis (55% v 20%, P < .02), severe ascites (64% v 13%, P < .001), and encephalopathy (45% v 3%, P < .001). We reach the following conclusions: (1) although not a frequent complication after distal splenorenal shunt in general, pancreatitis was commonly present in early postoperative deaths and was most likely a major contributor to the demise of those patients; (2) survivors with postdistal splenorenal shunt pancreatitis had a markedly increased morbidity rate; and (3) pancreatic complications after distal splenorenal shunt are more likely to occur in patients with pre-existing chronic pancreatitis. PMID- 9346539 TI - Liver transplantation for chronic viral hepatitis. PMID- 9346540 TI - Microchimerism in organ transplantation. PMID- 9346541 TI - A UNOS perspective on donor liver allocation. United Network for Organ Sharing. PMID- 9346542 TI - Who should receive the liver allograft: the transplant center or the recipient? PMID- 9346543 TI - The rational development of liver allocation policy. PMID- 9346544 TI - Liver transplantation in a managed care environment. PMID- 9346545 TI - What kind of a good is a donor liver anyway, and why should we care? PMID- 9346547 TI - CA19-9 does not predict cholangiocarcinoma in patients with primary sclerosing cholangitis undergoing liver transplantation. AB - The results of liver transplantation in patients with cholangiocarcinoma have been poor. It has been suggested that elevated serum CA19-9 levels predict cholangiocarcinoma in patients with primary sclerosing cholangitis. We analyzed the predictive value of CA19-9 antigen as a marker of cholangiocarcinoma in patients with primary sclerosing cholangitis evaluated for liver transplantation. We reviewed the charts of 26 patients with primary sclerosing cholangitis (stage IV) in whom preoperative serum CA19-9 levels were determined; 22 of 26 underwent liver transplant. Explant specimens were serially sectioned and examined for tumor. In 3 of the 26 patients, cholangiocarcinoma was diagnosed during pretransplantation evaluation; exploratory laparotomy on the last patient showed no evidence of cholangiocarcinoma, and this patient is awaiting transplantation. Twelve of the 26 patients had CA19-9 levels more than double the laboratory reference range (0-37 U/mL) (mean 183.1 +/- 103 U/mL, range 77-415 U/mL). Two of the 12 patients with elevated CA19-9 levels had cholangiocarcinoma. Of the 14 patients with normal levels, two had cholangiocarcinoma. No correlation between elevated CA19-9 and bile duct dysplasia was noted. Sensitivity for serum CA19-9 levels more than twice the reference range is 50%, specificity is 54.5%, positive predictive value is 16.6%. An elevated serum CA19-9 level in a patient with stage IV primary sclerosing cholangitis does not reliably predict coexisting cholangiocarcinoma. Persistently high or rising serum CA19-9 levels do not indicate more urgent need for liver transplantation. PMID- 9346546 TI - Differential patterns of reaction of human natural antibodies to pig hepatocytes and vascular endothelium. AB - We have recently conducted a series of experiments to characterize the pattern of reaction of human natural antibodies (NA) with individual pig liver cells. Pooled normal human serum (PHS) was incubated with cultured pig hepatocytes (HEP), aortic endothelial cells (AEC), and portal endothelial cells (PEC), and the reaction of NA to different cell types was measured by antibody-mediated cytotoxic (MTT assay), antibody binding (ELISA), and flow cytometric analysis. The human NA displayed a differential pattern of binding with hepatocytes exhibiting a more limited expression of xenoantigen expression than either aortic or portal endothelial cells. These differences in reaction patterns were also noted for Western blot analysis of individual cell membrane extracts. Preincubation of the pig cells with anti-pig MHC antibodies did not inhibit the binding of human IgM natural antibodies to the pig cells. Comparison of the pattern of NA absorption following the use of bioartificial liver support in patients with acute hepatic failure demonstrated limited ability of pig hepatocytes to absorb substantial amounts of NA. These studies indicate that pig hepatocytes are less vulnerable to NA cytotoxicity than pig vascular endothelial cells and that pig vascular endothelial cells express xenoantigens that are unique and not found on hepatocytes. PMID- 9346548 TI - Liver transplantation for treatment of giant hepatocellular adenomas. AB - Giant hepatocellular adenomas are associated with a high incidence of rupture with intra-abdominal hemorrhage and may also undergo malignant transformation. If resection is not technically feasible, liver transplantation should be a treatment option. The aim of this report is to describe the indications, feasibility, and outcome of liver transplantation for hepatocellular adenomas. A 66-year-old man with a 17-cm hepatocellular adenoma originating in the left lobe but involving nearly the entire liver and a 35-year-old woman with a 20-cm tumor involving the right lobe of the liver and compressing the left lobe underwent liver transplantation without complication. In both cases, a histological diagnosis was made by core needle biopsy preoperatively, and resection was technically not possible. Hepatocellular adenoma involving nearly all of the liver with no evidence of malignant change was confirmed in the explant liver from both cases. Giant hepatocellular adenomas may be unresectable and require liver transplantation for complete removal to prevent potential rupture with hemorrhage or malignant transformation. PMID- 9346549 TI - Primary malignant melanoma of the biliary tract. AB - Primary malignant melanoma of the biliary tract is an obscure entity, with only four previously reported cases. We report two cases involving the common bile duct. A 43-year-old male who underwent a right hepatectomy and excision of the extrahepatic biliary tree for a lesion at the bifurcation of the common bile duct. He remains alive and well 11 months after resection. The second patient is a 45 year old male with obstructive jaundice due to an ampullary lesion. Pancreaticoduodenectomy was performed with no signs of metastatic disease. He is 6 years following resection without evidence of disease. This is an unusual cause of obstructive jaundice and a definitive search for a possible extra-biliary primary should be pursued. In appropriately selected patients without evidence of metastatic disease, resection can potentially afford long-term survival if these lesions are true primary lesions and not metastatic from an undefined primary. However, given the high metastatic potential of melanoma it is unclear whether resection of these lesions results in cure or just effective long-term palliation. PMID- 9346550 TI - Overview: the management of metastatic carcinoid tumors. AB - Chemotherapy, IFN, octreotide, and hepatic artery embolization can be useful for palliation, both of local symptoms and of the carcinoid syndrome, but they have little effect on tumor progression, MIBG therapy is still experimental but may have a future role. OLT should be considered as a palliative procedure in those patients with severe symptoms and no evidence of extrahepatic disease. PMID- 9346551 TI - Metastatic endocrine tumors: is there a place for liver transplantation? AB - The authors describe their experience with liver transplantation (OLT) for metastatic endocrine tumors (MET) in order to determine reasonable indications for OLT in patients with this disease. Removal of the primary lesion and subsequent liver transplantation were performed in two separate procedures in all patients except one. Only those patients suffering from objective tumor progression and symptoms with no evidence of extrahepatic spread after complete work-up (including endoscopic ultrasonography (US) and 123I-labeled Tyr3 octreotide body scanning) underwent liver transplantation. Fifteen patients were referred for liver transplantation. Seven patients were excluded either because of stability of liver metastases (n = 3), extrahepatic spread, general contraindication (n = 2), or feasibility of aggressive surgical resection (n = 2). Liver transplantation was undertaken in eight patients with carcinoid tumor (n = 4), gastrinoma (n = 3) and glucagonoma (n = 1). Three patients did not survive the surgical procedure itself, whereas two additional patients died from chronic rejection or from recurrent disease. Three patients who received transplants for metastatic carcinoid tumor are alive without biochemical or imaging evidence of disease recurrence at 6, 15, and 52 months. The best indication for transplantation seems to be patients with metastases restricted to the liver and unresponsive to adjuvant therapy after aggressive surgical resection including excision of the primary lesion and reduction of hepatic metastases. In such highly-selected patients, liver transplantation remains a high-risk operation, but it can yield long-term survival. PMID- 9346552 TI - Orthotopic liver transplantation in the treatment of metastatic neuroendocrine tumors of the liver. AB - The place of orthotopic liver transplantation (OLT) in the management of metastatic hepatic neuroendocrine tumors has not been adequately defined. The present report is concerned with patient survival, disease recurrence, and symptom relief in 11 such patients in a single center who, at the time of transplantation, had no extrahepatic tumor. All patients obtained complete symptom relief initially but tumor recurrence was observed in 6 of the 11 cases (5 carcinoid and 1 apudoma) at a median of 11 months (range 3.5-26). Five patients have died, 4 in the carcinoid group with recurrence and one from chronic rejection in the other apudoma group. Of the 6 patients currently alive one of 2 carcinoids and one of 4 other apudomas have tumor recurrence. Recurrent deposits were found predominantly in bone and in the transplanted liver. Actuarial survival post transplant was 82% and 57% at 1 and 5 years respectively. It is concluded that OLT is effective at controlling symptoms from secreting carcinoid deposits in the liver. Although the tumor will recur in most cases, this is not necessarily associated with early return of symptoms. Prolonged disease free survival is more likely in the non carcinoid apudoma group. PMID- 9346553 TI - Carcinoid tumors of the extrahepatic bile duct: an unusual cause of bile duct obstruction. PMID- 9346554 TI - Reperfusion injury to donor livers stored for transplantation. PMID- 9346555 TI - Hepatocyte transplantation. PMID- 9346556 TI - Biliary reconstruction for liver transplantation and management of biliary complications: overview and survey of current practices in the United States. PMID- 9346557 TI - Overview: biliary reconstruction after liver transplantation. PMID- 9346558 TI - Malignant vascular tumors of the liver presenting as liver failure and portal hypertension. AB - We describe three patients referred for orthotopic liver transplantation with liver failure and portal hypertension who were found to have malignant vascular tumors: two patients with angiosarcoma and one patient with epithelioid hemangioendothelioma. Their clinical presentation mimicked decompensated chronic liver disease. None had tumor masses on computed tomography and ultrasonography. Massive tumor involvement of the liver was identified in the two patients studied by magnetic resonance imaging. Pathological examination of the explanted liver at the time of transplantation in one patient and autopsy in a second patient showed angiosarcoma. Laparoscopic liver biopsies in the third patient showed epithelioid hemangioendothelioma. The vascular origin of the tumor was established by histopathologic examination and confirmed with immunohistochemistry. Malignant vascular tumors of the liver should be included in the differential diagnosis of liver failure of unclear etiology. PMID- 9346559 TI - Recurrence of autoimmune hepatitis following liver transplantation. PMID- 9346561 TI - Acute liver failure and transplantation: a symposium. PMID- 9346560 TI - Hepatitis C--associated glomerular disease in liver transplant recipients. AB - Hepatitis C virus (HCV) infection may be associated with extrahepatic illness including renal disease. We investigated the clinical and virological characteristics of three patients who developed a mesangial proliferative and sclerosing glomerulopathy alone or in association with membranoproliferative glomerulonephritis after liver transplantation for end-stage liver disease secondary to HCV infection. Using polymerase chain reaction technology and the IgM RIBA assay, viral load, genotype and IgM antibody response to HCV in the setting of glomerulonephritis was evaluated. Within 1 year of transplantation, the patients showed decreased renal function, proteinuria and recurrent hepatitis C liver disease. Likewise, HCV viral load increased following transplantation, whereas the viral genotypes remained unchanged. Although the first patient presented with classic type II cryoglobulinemia in association with glomerulonephritis, the second patient developed an IgM directed specifically against the hepatitis C core antigen. The third patient developed a low-titered IgM directed against the hepatitis C core antigen with rheumatoid factor activity but without cryoglobulinemia. All of the patients show IgM in glomerular capillary walls by biopsy. One patient has shown a clinical response to interferon (IFN) alfa-2b therapy without evidence of hepatic allograft rejection. The second and third patients have not responded to IFN or developed hepatic rejection. This study suggests that HCV-associated glomerulonephritis may complicate liver transplantation in conjunction with the production of increased amounts of IgM of variable specificity. The posttransplant setting may provide a unique situation in which to investigate the specific requirements for the onset of renal disease. PMID- 9346562 TI - Liver transplantation for fulminant hepatic failure: North American experience. PMID- 9346563 TI - Timing and candidacy for transplantation in acute liver failure: the European experience. PMID- 9346564 TI - Cerebral edema and intracranial pressure monitoring. AB - With the wide acceptance of liver transplantation as a therapeutic alternative in fulminant hepatic failure (FHF), the successful management of patients with this syndrome has acquired a new urgency. Topping the list of medical problems is the development of brain swelling. Two decades after the recognition of its importance, brain edema and intracranial hypertension still constitute a major cause of death in these patients. In a more recent classification of FHF, brain edema was especially prominent in those subjects with "hyperacute failure," in whom a period of 7 days or less elapsed between the development of jaundice and encephalopathy. The goal of this review is to discuss two aspects of this clinical problem. On one hand, elucidation of its pathogenesis should lead to a more rational therapeutic approach; such an information would also be valuable to understand the relationship between hepatic encephalopathy and brain edema, a source of controversy. Studies of pathogenic mechanisms are difficult to perform in humans and animal models of FHF have proven valuable, as brain swelling can be detected with some regularity. On the other hand, an increasing array of techniques is now available in the intensive care setting to monitor patients with FHF. Of these, intracranial pressure monitoring has received the most critical attention. However, concerns with the risks of craniotomy and the need to acquire more dynamic information has led several groups to explore non invasive methods that evaluate the consequences of intracranial hypertension. Their role, though potentially exciting, is still uncertain. PMID- 9346565 TI - Auxiliary liver transplantation for acute liver failure. AB - In summary, the following principles are worth reiterating: 1. In the treatment of acute liver failure, protection of the native liver in anticipation that it will recover, but positioning of the allograft in a manner that optimizes its function for both the short and long term (in the event that the native liver does not recover) are important goals. Therefore, orthotopic positioning offers advantages over the heterotopic position in most cases. Development of better techniques for predicting native liver recovery might remove any of these advantages of the orthotopic position. 2. Other than the presence of fibrosis, the performance of a native liver biopsy does not appear to predict native liver recovery. The decision of whether to attempt auxiliary grafting must be based on an understanding of the natural history of the disease causing the acute liver failure. 3. The heterotopic position has the advantage of not requiring partial native hepatectomy in order to accommodate the allograft. However, except for the recent experience of Terpstra et al, this technique has carried a higher risk of venous outflow obstruction. It also requires additional space within the abdomen, usually mandating the use of prosthetic abdominal wall closures and the construction of venous conduits for portal venous inflow to the liver. There is the additional theoretical concern about competition for portal venous flow leading to eventual atrophy of the allograft liver. 4. Common events that follow liver transplantation result in changes in portal venous resistance within the liver, events that therefore alter the relative distribution of portal venous inflow between native and auxiliary livers. These events include reperfusion injury, allograft rejection, allograft viral infection (e.g., cytomegalovirus, Epstein-Barr virus, recurrent viral hepatitis), and native liver regeneration. Attempts to control portal venous flow to favor one liver over the other must account for the effect of these factors. 5. In general terms, auxiliary transplantation is not indicated for diseases in which the residual native liver either represents an ongoing threat to the recipient or is incapable of supporting life alone. This may be the case in both metabolic disorders and in cirrhosis. Most of the alleged difficulties of native hepatectomy are no longer relevant. Therefore, auxiliary transplantation is rarely if ever indicated for chronic liver disease and may not be of any additional benefit over total transplantation in the treatment of many metabolic disorders. 6. In the treatment of acute liver failure, the value of an auxiliary transplant over total transplant is obtained when the native liver recovers and the patient is withdrawn from immunosuppression. If further experience shows the effectiveness of this option, total liver transplantation with the requirement for life-long immunosuppression will no longer be appropriate for the treatment of patients with acute liver disease. PMID- 9346566 TI - Evaluation of extracorporeal bioartificial liver devices. AB - The initial clinical studies on artificial liver support were performed at King's College Hospital, London during the late 1970s. Initially using charcoal haemoperfusion, and subsequently resin haemoperfusion, and these studies culminated in a controlled clinical trial of charcoal haemoperfusion in which overall survival was high, but no statistically significant benefit was found. From this study, much information was also obtained about the clinical importance of the various complications of acute liver failure, and the experience of King's over the last 2 decades exceeds 1,000 patients. The aim of this article is to review the potential of the exciting new developments in this field of bioartificial liver support incorporating hepatocytes. It focuses on the published findings of early clinical use of these systems and attempts to identify what is needed in further studies. It is of paramount importance that future trials are designed to give the greatest information on the effects of bioartificial liver support. For these, the biocompatibility of the systems should to be confirmed with detailed assessment and sensitive tests need to be developed to determine the metabolic functional efficacy of these devices. The possible relationship of treatment to liver regeneration has been considered, because without this these systems will in many cases be limited to use as a bridge to liver transplantation. Finally, some of the possible future modifications of the cell-based liver support systems are discussed. PMID- 9346567 TI - Surf's up. PMID- 9346568 TI - Acute liver failure: results of a 5-year clinical protocol. AB - This investigation summarizes and evaluates the results of a clinical protocol that we designed to care for patients with acute liver failure (ALF). Adult patients with ALF were enrolled in the protocol. Grade II portal-systemic encephalopathy prompted admission to the intensive care unit (ICU). Patients who met the clinical criterion were activated for liver transplantation. Intracranial pressure (ICP) was monitored in patients with grade III encephalopathy. An increase in ICP was treated with hyperventilation, diuretics, barbiturates, or a combination thereof. Survival was considered to have occurred if the patient left the hospital alive. Our series included 25 patients. Orthotopic liver transplantation (OLT) was performed on 19 patients, 12 of whom survived. Only 2 of 6 patients who did not undergo transplantation survived. Ten of 11 patients who underwent transplantation before reaching grade IV encephalopathy survived. Only 2 of 8 patients who underwent transplantation after reaching grade IV survived (P = .006). The causes of death included cerebral edema (3 patients), disseminated aspergillosis (3 patients), and other (5 patients). ICP was monitored in 11 patients. Increased pressure was documented by seven of the monitors placed. There was one focal hemorrhage secondary to a subdural monitor. Outcome is improved if transplantation occurs before grade IV encephalopathy. ICP monitoring can be accomplished without significant risk of hemorrhage. In our series, infection with aspergillus occurred frequently and with fatal outcome. PMID- 9346570 TI - Defining the role of transjugular intrahepatic portosystemic shunts in the management of portal hypertension. PMID- 9346571 TI - Transjugular intrahepatic portosystemic shunts: impact on liver transplantation. AB - This study was designed to evaluate the impact of transjugular intrahepatic portosystemic shunts (TIPS) on liver transplantation. Historically, the complications of portal hypertension have been temporized with sclerotherapy or surgical portosystemic shunts. In patients whose liver disease progressed, liver transplantation has been used as definitive treatment. More recently, TIPS is being used increasingly for the management of the complications of portal hypertension. The impact of this new modality on liver transplantation is evaluated. The records of 135 adult patients undergoing liver transplantation at University of California at Los Angeles between October 1992 and June 1993 were reviewed. Twenty-three patients had received at least one shunt before transplantation. The TIPS procedure complicated the operative course of 5 patients (22%). In 2 patients the TIPS had been placed cephalad, making placement of the suprahepatic vena caval clamp difficult. In 2 other patients, the shunt had been placed caudad, extending in the extrahepatic portal vein. In all 4 of these patients, the intima had been damaged at the area of the subsequent anastomosis. In the fifth patient, the bile duct had been perforated during the placement of the shunt, causing diffuse bile peritonitis, which was sterile, and the transplantation was performed. The average intraoperative blood loss for these 5 patients was 13 U. There was no significant decrease in intraoperative blood loss for all patients with a TIPS when compared with 112 adults who underwent liver transplantation during the same period (11 U v 10.5 U). The TIPS stent did not improve objective intraoperative parameters as compared with liver transplant recipients without TIPS. The indications for TIPS must be carefully weighed against the potential risks of increasing the technical difficulty of the transplantation and jeopardizing the candidacy of some liver transplantation candidates. Liver transplantation is not facilitated by TIPS insertion and therefore should not be used to justify TIPS placement. PMID- 9346572 TI - Liver transplantation for hemochromatosis: an ironic dilemma. PMID- 9346569 TI - Energy status in anoxic rat hepatocytes: effects of isoflurane, solution composition, and hypothermia. AB - Both cold and warm ischemia occur during liver transplantation. Hypothermia and Wisconsin solution preserve adenine nucleotide energy status, which is crucial to hepatic function and viability. The volatile anesthetic isoflurane has been shown to preserve energy status in anoxic isolated hepatocytes in warm Krebs solution. The present study examined isoflurane effects on energy status during incubation also in Wisconsin or Krebs-plus-adenosine solution at 37 degrees or 4 degrees. Hepatocytes were isolated from rat liver after perfusion with Krebs + collagenase. In 25-mL flasks, 12.5 million cells in 2.5 mL of Krebs, Krebs plus 5 mmol/L adenosine, or Wisconsin solution were incubated under an atmosphere of O2/CO2 or N2/CO2 (19:1) +/- isoflurane (3 volumes% = 2ED50), for 30 minutes at 37 degrees C or 4 degrees C. Adenine nucleotides were measured by high-performance liquid chromatography (HPLC), lactate enzymatically. During warm (37 degrees) anoxia, Wisconsin solution preserved energy status; Krebs plus adenosine did not. Isoflurane further protected energy status in all three solutions. Hypothermia (4 degrees) alone greatly decreased anoxic loss of energy status in all solutions. In Wisconsin solution only, energy status tended to be higher in anoxic than in oxygenated cells and was further enhanced by isoflurane, with corresponding increases in lactate. During 30 minutes of either warm or cold anoxia, isoflurane and Wisconsin solution each helped preserve adenine nucleotide energy status in isolated hepatocytes, at least in part through enhanced glycolysis. PMID- 9346573 TI - Primary liver cancer and survival in patients undergoing liver transplantation for hemochromatosis. AB - Cirrhotic patients with hereditary hemochromatosis (HHC) have an increased risk of primary liver cancer (PLC). The purpose of this study was to determine the prevalence of primary liver cancer in patients with HHC undergoing orthotopic liver transplantation (OLT). Five liver transplant centers were surveyed; clinical and pathological data on 37 patients with HHC undergoing OLT were retrospectively collected and analyzed. The diagnosis of HHC was established by a combination of serum transferrin-iron saturation, hepatic iron index (HII), and/or pattern of liver iron staining. The diagnosis of HHC had been unsuspected before OLT in 13 of 37 (35%). Primary liver cancer was found in the explants of 10 of 37 patients (27%) and was unsuspected in 7 of 10 (70%); 8 were hepatocellular carcinoma, and 2 were cholangiocarcinoma; foci of hepatocyte dysplasia were found in 6 additional patients. Mean (+/- SEM) hepatic iron content and HII in 20 patients without prior phlebotomy or bleeding were 17.2 mg/g dry weight (+/- 2.9) and 5.5 (+/- 0.8), respectively. The overall 1-year survival rate after OLT in the 37 HHC patients was 58% (v 55% for HHC patients with PLC). We draw the following conclusions: (1) the diagnosis of HHC is often unsuspected before OLT, and HHC should be evaluated pretransplantation by direct and indirect markers; (2) HHC patients undergoing OLT have a high prevalence of primary liver cancer, the majority being unsuspected; and (3) HHC patients have poorer than average survival after OLT, which cannot be explained solely by the presence of concomitant PLC. PMID- 9346574 TI - Liver transplantation for hepatocellular carcinoma: results with preoperative chemoembolization. AB - At the University of California, San Francisco, 17 patients who met the following criteria-hepatic tumor unresectable because of location or inadequate liver reserve, no metastases, HBsAg negative, no tumor larger than 5 cm in diameter, and no more than three tumors--were enrolled prospectively in a protocol employing preoperative chemoembolization to assess whether orthotopic liver transplantation (OLT) could cure a majority of highly selected patients with hepatocellular carcinoma (HCC). Thirteen patients had biopsy-proven HCC, 2 had the fibrolamellar variant, and 2 had radiological findings of HCC but no biopsy confirmation. Fourteen had underlying liver disease. All arteriographically apparent lesions were chemoembolized using a mixture including Gelfoam powder, doxorubicin, mitomycin-c, and cisplatin. Eight patients with poor hepatic reserve were chemoembolized when a donor organ became available, whereas 9 patients were chemoembolized and then placed on the waiting list. The only complication of chemoembolization was a gangrenous gallbladder in 1 patient. Thirteen patients underwent liver transplantation (2 patients without prior histological confirmation of carcinoma had no identifiable tumor at OLT); 3 patients developed metastases between the time of enrollment and donor organ availability and subsequently died; and 1 patient underwent a trisegmentectomy. Ten of the 11 patients with biopsy-proven HCC who underwent transplantation remain free of recurrent cancer at a median of 40 months; 1 patient died at 6 months of lymphoproliferative disease with no cancer found at autopsy. Although the role of chemoembolization is uncertain, these data show that the majority of carefully selected patients with HCC may achieve long-term survival with OLT. PMID- 9346575 TI - The role of tumor markers in the diagnosis of hepatocellular carcinoma, with special reference to the des-gamma-carboxy prothrombin. AB - The assessment of new and more sensitive serum markers for hepatocellular carcinoma (HCC) represents a useful contribution to the diagnosis of small liver tumors, still amenable by surgery. We evaluated the efficacy of the tumor markers proposed during recent years for the study of HCC: alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), serum ferritin (SF), tissue polypeptide antigen (TPA), and, finally, the more recently proposed des-gamma-carboxy prothrombin (DCP). Of the 227 patients included in this retrospective study, 111 had HCC, and 85 of these were also cirrhotic. The remaining 116 patients, considered as the control group, included 23 patients with liver metastases from colorectal cancer, 26 with benign hepatic lesions, 20 with tumors other than HCC without hepatic metastases, and 47 with other liver diseases. For each single tumor marker, the sensitivity, specificity, positive and negative predictive values, diagnostic accuracy, and Younden index were assessed. AFP and DCP proved to be the most effective, with sensitivity, specificity, and diagnostic accuracy of 54.9%, 97.4%, and 76.6% and of 53.3%, 88.1%, and 71.1%, respectively. The same parameters evaluated for combined use of the two markers were 74.2%, 87.2%, and 80.9%, respectively. Analysis of the other markers produced no further significant contribution. Of the 111 patients with HCC, 35 (33.3%) were positive for both AFP and DCP, 43 (41%) were positive for one of them, and 27 (25.7%) were completely negative. In the 44 patients who underwent liver resection or transplantation, DCP correlated significantly with the histological presence of microvascular thrombosis, the major factor determining long-term survival after curative surgery. As a tumor marker for HCC, DCP is at least as effective as AFP; the combined use of AFP and DCP significantly improves the chances of identifying HCC by serodiagnosis. PMID- 9346576 TI - Primary sclerosing cholangitis: liver transplantation or biliary surgery. PMID- 9346577 TI - Amelioration of acute GVHD disease and reestablishment of tolerance by short term treatment with anti TCR antibody. PMID- 9346578 TI - Liver transplantation for hepatitis B. PMID- 9346579 TI - Should liver transplantation be performed for patients with chronic hepatitis B? Yes! AB - The use of passive immunoprophylaxis to prevent HBV reinfection of the allograft following liver transplantation has led to a dramatic improvement in the outcome of patients who undergo transplantation for chronic hepatitis B. Hepatitis B previously was not one of our favorite diseases for which to perform liver transplantation. However, we now regard patients with this disease as good candidates for liver transplantation. I hope I have presented a compelling argument that no patient with hepatitis B, regardless of serological status, should be a priori denied liver transplantation. Studies from both the U.S. and Europe have shown that HBV reinfection can be prevented in almost all patients at a cost not out of line with other accepted indications for liver transplantation. Thus, I once again, ask the question, "Should patients with chronic hepatitis B undergo liver transplantation?" At our institution, the answer is an emphatic Yes! PMID- 9346580 TI - Liver transplantation for hepatitis B: the con aspect. PMID- 9346581 TI - Liver transplantation for hepatitis B virus infection. PMID- 9346582 TI - Liver transplantation for hepatitis B: what have we learned and what does the future hold? PMID- 9346583 TI - National Transplantation Pregnancy Registry: analysis of pregnancy outcomes in female liver transplant recipients. AB - Outcomes from 48 pregnancies in 34 female liver transplant recipients were analyzed. Data were collected via interviews, questionnaires, and hospital records. All recipients were treated with cyclosporine-based immunosuppression except 2 patients treated with FK506 and 2 treated with no immunosuppression. The age at conception was 26.1 +/- 5.9 years (mean +/- SD) with a transplant interval (time from transplantation to conception) of 2.9 +/- 2.5 years. There were 49 outcomes (1 set of twins): miscarriage 9 (18%), therapeutic abortion 4 (8%), and live birth 36 (74%). No stillbirths or ectopic pregnancies were reported. Of the 36 live births, the gestational age was 36.9 +/- 3.5 weeks, the birthweight was 2,604 +/- 698 grams, 39% were premature (< 37 weeks), and 31% had low birthweight (< 2,500 grams). No birth defects or neonatal deaths (< 28 days) were reported. The newborn complication rate was 17% (n = 6), 5% in premature infants. The incidence of drug-treated hypertension was 46%; pre-eclampsia 21%; infectious complications 26%; and Caesarean section 47%. Recipients with hypertension had a higher proportion of premature infants (71%) than normotensive patients (38%) (P = .04 by Fisher's exact test). Acute rejection was diagnosed in 6 pregnancies, 2 of which were ended by therapeutic abortion. Four recipients who continued their pregnancies were treated with increased immunosuppression for rejection, and all delivered livebirths. There were two grafts lost within 6 months of pregnancy. The only maternal death occurred in a patient who required retransplantation for recurrent C hepatitis 3 months afte therapeutic abortion and died 6 months later. The other recipient with graft loss was successfully retransplanted for chronic rejection 6 months after delivery. We draw the following conclusions: (1) female liver transplant recipients can safely undergo pregnancy, although there is a high rate of premature and low birthweight infants; (2) pregnancies in this population should be considered high-risk and require close monitoring of liver function; and (3) altered graft function during pregnancy should be thoroughly investigated. PMID- 9346584 TI - Centrilobular necrosis after orthotopic liver transplantation: a longitudinal clinicopathologic study in 71 patients. AB - Centrilobular necrosis (CLN) is a histological finding often encountered after orthotopic liver transplantation, but its pathogenesis is still unknown. In this study, the significance of CLN was assessed in a series of 227 consecutive liver transplantations performed between January 1989, and December 1991. Seventy-one patients (30.9%) showed CLN on at least one biopsy result, which were obtained because of an increase of aspartate aminotransferase activity. Their liver specimens were reviewed, and 19 histological features were recorded with particular attention given to lobular changes in acinar zone 3, to features commonly attributed to cellular and ductopenic rejection, and to changes suggestive of ischemia. CLN could first be observed either soon (within 4 days) or late (up to 3 years) after transplantation. Only 23 (32.4%) specimens had centrilobular necrosis affecting more than 75% of acinar zones 3. In 60 cases (84.5%) the lesion was limited to acinar zone 3. An important associated feature was sinusoidal congestion in 73.2% of cases. Fifty-one of 71 patients (71.8%) had histological features of cellular rejection before or at the time of CLN, and 13 of these progressed to ductopenic rejection versus 3 of the 156 patients without CLN (P < .0001). Nine patients had a recurrence of CLN, of whom 2 progressed to ductopenic rejection, a recurrence rate of 16.7% in this series. The survival of patients with CLN is worsened by associated ductopenic portal tracts compared with those without ductopenia (P = .0189-Mantel-Cox). This histological combination, irrespective of the serum bilirubin level, may warrant an early conversion to FK506-based immunosuppression. PMID- 9346585 TI - Hepatic allograft rejection is associated with increased levels of soluble intercellular adhesion molecule-1. AB - Intercellular adhesion molecule-1 (ICAM-1) is an adhesion molecule from the immunoglobulin super family that is recognized to be an important factor in the multistep process of cell transendothelial migration and lymphocyte adhesion during antigen recognition and effector cytolysis, mechanisms known to be involved in the pathogenesis of hepatic allograft rejection. A soluble form of ICAM-1 (sICAM-1) can be shed into the circulation. In this study, we examined the levels of sICAM-1 in hepatic allograft recipients as possible markers of cellular rejection and the presence of cytomegalovirus (CMV) hepatitis. We studied three groups of patients, including eight patients with histologically documented cellular rejection, five patients with histologically documented CMV hepatitis, and a liver transplantation control population. Serum samples were obtained at the following times: baseline (1 to 3 days after transplantation), at time of diagnosis of cellular rejection, and at time of diagnosis of CMV hepatitis and 1 week after treatment of rejection episodes. The levels of sICAM-1 were measured using an established commercial enzyme immunoassay with a sensitivity of 0.3 ng/mL. We found that serum levels of sICAM-1 were significantly increased in liver transplant recipients who were experiencing hepatic allograft rejection but were unchanged in patients with CMV hepatitis or the time-matched liver transplant controls. Serum levels of sICAM-1 decreased significantly after successful treatment of the rejection episode with bolus corticosteroid therapy. We conclude that serum levels of sICAM-1 may be useful in monitoring the occurrence of rejection and the response to antirejection therapy in liver transplant recipients. PMID- 9346586 TI - Contribution of true cold and rewarming ischemia times to factors determining outcome after orthotopic liver transplantation. AB - The role of true cold ischemia times (CIT) and rewarming ischemia times (WIT) in determining outcome after liver transplantation was investigated in 230 adult recipients. Using multivariate analysis, WIT (time from the start of implantation until restoration of arterial and portal blood supply) and donor intensive care stay (P = .04 and .0004, respectively) but not CIT (the time from donor portal vein flushing until the graft was removed from University of Wisconsin solution; P > .30) emerged as independent determinants of graft survival. In the small number of patients with a WIT of greater than 180 minutes, there were reductions in graft survival (58% v 80% for WIT greater than 180 minutes) but these just failed to reach significance (P = .055). CIT had no influence on graft survival using cut-offs of 12 or 18 hours. A WIT of greater than 180 minutes was associated with an increased median area under the curve of day 1 through 7 serum bilirubin (1,370 v 915 mumol/L.day; P = .048) and trends towards an increased incidence of primary graft nonfunction or dysfunction (22.2% v 6.2% for WIT of less than 180 minutes; P = .065) and the day 1 through 7 area under the curve of serum aspartate aminotransferase (3,310 v 1,440 IU/L.day; P = .092). A prolonged CIT (greater than 18 hours) led to a prolonged hospital stay (69 v 31 days; P = .03), an increased area under the curve of day 8 through 14 serum bilirubin (2,500 v 995 mumol/L.day; P = .003), and a trend towards an increased incidence of initial poor graft function (33.3% v 6.3% for less than 18 hours; P = .092). The incidence of acute rejection increased (to 64.3% from 53.4%; P = .04) in patients with preservation injury (serum aspartate aminotransferase greater than 1,500 IU/L during the first 2 postoperative days). True CIT and WIT are important determinants of outcome after liver transplantation. PMID- 9346587 TI - Hepatic ischemia-reperfusion injury modification during liver surgery in rats: pretreatment with nifedipine or misoprostol. AB - The aim of the study was to determine if pretreatment with misoprostol (a prostaglandin analogue) or nifedipine (a calcium antagonist), know protectants of the whole liver, would ameliorate the ischemia-reperfusion injury (IRI) of resected liver associated with vascular occlusion. Male Wistar rats were allocated to 5 groups (n = 20 each group): sham-operated, liver resection only, liver resection plus pretreatment with 0.1 mg/kg misoprostol, 10 mg/kg, or 2 mg/kg nifedipine during the 3 days before IRI with liver resection. Fifteen percent of the liver was made ischemic by 30-minute continuous vascular occlusion, and the remaining 85% nonischemic liver was resected. The model was designed to have survival of the rats so that liver function could be studied over 3 weeks. Seventeen of 20 control resection rats survived indicating a suitable model for study. The bilirubin level was reduced by 25% on postoperative days 3 through 23 with misoprostol. The serum alanine aminotransferase (ALT) peak was significantly lower on day 1 with misoprostol and high-dose nifedipine (both reduced to half the control resection value). There was a modest but significant reduction of serum alkaline phosphatase (SAP) for low-dose nifedipine on days 1, 2, and 23. Prothrombin had a lower peak and lower values on days 1 through 4 with misoprostol. Liver histological changes were minor, being cytoplasmic vacuolization only, and was slightly more marked in the nifedipine groups. Preoperative misoprostol 0.1 mg/kg and nifedipine 10 mg/kg each ameliorate the IRI associated with liver resection, as measured by liver function tests. Different aspects of liver function were altered by the different agents. These results justify initiating a trial for human liver resections. PMID- 9346589 TI - Issues in xenotransplantation. PMID- 9346588 TI - Influence of donor age on graft survival after liver transplantation--United Network for Organ Sharing Registry. AB - The waiting list for liver transplantation has more than doubled between 1988 and 1992, yet the number of liver transplantations during the same period increased by only 79%. This discrepancy is due to the limited availability of donors. The modest increase in donor pool is caused entirely by donors > or = 40 years of age, a trend likely to continue. To determine the impact of increasing donor age on the outcome of liver transplantation, we analyzed 6-month graft survival in 7,988 adults who received first liver graft between October 1987 and September 1992, and were observed through the United Network for Organ Sharing Scientific Liver Transplant Registry. Graft survival was measured by death and/or retransplantation, donor age by decades. The independent effect of donor age on graft survival was estimated by Cox regression analysis controlling for the possible confounding of donor, recipient, and institutional characteristics. Between 1987 and 1992, the percentage of donors over 50 years increased from 2.1% to 17.5% resulting in change of median donor age from 23 to 31 years. For donor age > or = 50, graft failure rate was 50% higher than with donor age less than 20 years (excess for mortality was 25% and for retransplantation 94%). Adjustment for characteristics associated to donor age or outcome did not eliminate the excess risk found with increasing donor age. Despite these adversities, graft failure rate in recipients from oldest donors (27.2%) in 1992 was nearly equivalent to recipients from the youngest donors (26.9%) in 1987 to 1988. Although increasing donor age has an adverse effect on 6-month graft survival, improvement in transplantation technology and patient care over time have more than compensated for poorer graft function associated with the simultaneous rise in median donor age. PMID- 9346590 TI - General considerations in the management of patients with depressed consciousness and chronic hepatic insufficiency. PMID- 9346591 TI - Impaired consciousness after liver transplantation. PMID- 9346592 TI - Seizures after liver transplantation. PMID- 9346594 TI - The value of P. PMID- 9346593 TI - Age and liver donation. PMID- 9346595 TI - Immunosuppression in liver transplantation: monitoring, dose adjustment, reduction, and withdrawal. PMID- 9346596 TI - Nephrotoxicity associated with cyclosporine and FK506. AB - In conclusion, nephrotoxicity is a common and potentially clinically significant problem with cyclosporine and FK506 following transplantation. Further definition of its pathogenesis and improved dosing strategies for both of these drugs may reduce the impact of nephrotoxicity on both short-term and long-term outcomes. PMID- 9346598 TI - Hyperlipidemia and other coronary risk factors after orthotopic liver transplantation: pathogenesis, diagnosis, and management. PMID- 9346597 TI - Hypertension after liver transplantation. AB - Hypertension developing after liver transplantation is nearly universal and likely reflects several pathogenic mechanisms. Foremost among these are altered vascular reactivity and vasoconstriction related to CSA, and probably FK506, administration, impaired GFR and sodium excretion, and the effects of steroids. This disorder is of both theoretical and practical importance in understanding blood pressure regulation in humans. Most importantly, it poses a considerable long-term cardiovascular risk for the transplant recipient. Recognition of acquired hypertension and timely intervention are among the primary management challenges for the transplant clinician. PMID- 9346599 TI - Neurological and psychiatric complications associated with immunosuppression in liver transplantation. PMID- 9346600 TI - Long-term management of biliary tract complications. AB - Biliary tract complications represent an important problem after transplantation. They are not only important in the immediate posttransplantation period, but perhaps of even greater importance in the long-term management of transplant recipients. One of the major changes that has occurred over the past several years is an increasing use of nonoperative therapy, either via endoscopic or percutaneous routes. In many cases of biliary leak, stones, and T-tube malposition, nonoperative management may be curative. The exact role they play in the management of strictures, particularly diffuse stricturing and isolated hepatic duct strictures, remains to be determined. At the least, it provides temporary drainage before initiating surgical therapy. In addition to being efficacious, nonsurgical management is quite safe. In 64 cases of biliary complications managed nonoperatively, complications occurred at UCSF in only four patients. One patient developed bleeding along the percutaneous catheter tract and required replacement with a larger diameter catheter to tamponade the bleeding. Two patients developed hemobilia secondary to intrahepatic pseudoaneurysms caused by the percutaneous drainage catheter. One patient developed an osteomyelitis of a rib related to an indwelling percutaneous biliary catheter that responded to antibiotics and rib resection. Finally, understanding several of the biliary complications that occur after transplantation (ie, sphincter of Oddi dysfunction) will require additional research and hopefully will determine the pathophysiology of these complications. This should lead to more rational therapy. PMID- 9346601 TI - Bone disease after liver transplantation. PMID- 9346602 TI - The effect of immunosuppression on growth and development. PMID- 9346603 TI - Sexuality of older women after liver transplantation: interrelationships of prednisone use, sex steroid hormone levels, body image, and intimacy. PMID- 9346604 TI - Parenthood after liver transplantation. AB - In conclusion, data from the NTPR support the concept that female liver transplant recipients can safely undergo pregnancy, and male recipients are able to father pregnancies. The true incidence of malformations in both populations needs to be further studied. Female liver recipients have a high rate of premature and low-birth-weight infants. Therefore, pregnancies in the female population must be considered high risk and require close monitoring of liver function. Our data and other reports suggest that altered graft function during pregnancy may represent rejection and must be thoroughly investigated. Data from female CsA kidney recipients and from case reports of liver CsA recipients would suggest that increased doses of CsA may be required during pregnancy. The confounding effects of renal function, hypertension, and combinations of drugs make the interpretation of newborn outcomes and the relationship to immunosuppressive regimens difficult to interpret. To date, an increase in congenital anomalies has not been reported in newborns of liver recipients. Therefore, although subtle effects on reproduction may occur, it seems that favourable pregnancy outcomes can be expected for most liver transplant recipients, but further study is needed as the sample size has been small. PMID- 9346606 TI - Epstein-Barr viral infection and posttransplantation lymphoproliferative disorders. PMID- 9346605 TI - Recurrence of viral hepatitis after liver transplantation: insights into management. AB - Recurrence of viral infection and hepatitis is a common problem for patients undergoing LT for hepatitis B or hepatitis C. In patients with hepatitis B who do not receive immunoprophylaxis, recurrence of HBsAg positivity is virtually universal and is usually associated with rapidly progressive hepatitis that jeopardizes long-term patient and allograft survival. HCV infection recurs in 80% to 100% of patients, but only 50% develop histologic features of hepatitis, which are generally mild and do not significantly decrease life survival. Long-term HBIg prophylaxis is currently the only effective strategy to prevent or modify HBV recurrence. At present, there is no effective prophylaxis for recurrence of HCV infection. Preliminary results suggest that interferon therapy may benefit a minority of patients with either recurrent HBV or HCV infection after LT. Hepatitis B should not be regarded as a contraindication for LT. However, until an effective and readily available therapy is developed to prevent recurrence, HBsAg-positive patients should undergo transplantation under experimental protocols. Hepatitis C is also not a contraindication for LT. Although recurrent hepatitis C is usually mild and slowly progressive, severe forms of hepatitis requiring retransplantation have been increasingly reported. Long-term follow-up studies are needed to define the natural history of recurrent HCV infection after LT and its impact on allograft and patient survival. PMID- 9346607 TI - Recurrence of primary biliary cirrhosis, primary sclerosing cholangitis, and autoimmune hepatitis. AB - Thus, controversy remains as to whether PBC or PSC does recur in the allograft. Both diseases are of unknown origin. If PBC does recur in the allograft then, on current evidence, it is likely to recur only in a minority of patients at least in the short to medium term. Furthermore, recurrent does not appear to impact on patient or graft survival. As with chronic hepatitis B, the degree of immunosuppression may also affect the pattern and rate of disease recurrence. Sclerosing cholangitis does affect the graft, but the clinically severe form appears to be related to factors other than recurrent disease. The histological features of PSC are nondiagnostic but one study at least does suggest that, as with PBC, the pattern of disease recurrence is patchy and early in the medium term. Autoimmune hepatitis may recur but the patterns of recurrent disease are uncertain. If some of the reports do show recurrent disease, then it seems that recurrent disease is more aggressive in the allograft than in the native liver. Disease recurrence is of more academic and practical importance. Clinicians need to be aware of possible recurrence and, where appropriate, modify immunosuppression and interpret liver tests and histology appropriately. Finally, the patterns of recurrence may give useful clues to the pathogenesis of these enigmatic diseases. PMID- 9346608 TI - Social, vocational, and financial consequences of liver transplantation. PMID- 9346609 TI - Long-term follow-up of the liver transplant recipient. PMID- 9346610 TI - The relationship between outcome of liver transplantation and experience in new centers. AB - For several medical interventions, increasing experience results in improved outcome. This finding may result from better patient selection or increased skill levels. This report examines whether there is a relationship between center experience and patient outcome for liver transplantation, and if so, whether the relationship is explained by patient or donor selection or level of experience required to obtain optimal results. The United Network for Organ Sharing Scientific Liver Transplant Registry includes all procedures performed in the United States since October 1987. The date of the first transplantation and the number of operations performed were used to define 42 new and 27 experienced centers. Within new centers, experience was quantified by the sequence number of each transplantation. Characteristics of 6,180 recipients and donors were compared between new and experienced centers using the chi 2 test for association. A linear trend test identified whether these characteristics varied with experience within new centers. The independent association between experience in new centers and perioperative mortality was examined using logistic regression. Patient and donor selection criteria differ between experienced and new centers and change within new centers as experience is gained. Adjusting for calendar year and various patient and donor characteristics, perioperative mortality rates decrease in new centers as experience is gained. After 20 transplantations are performed, perioperative mortality in new centers is not significantly different than that in experienced centers. Criteria for recipient and donor selection change as centers gain experience. Despite these differences and improvements that have occurred over time, increasing experience in centers performing liver transplantations is associated with reduced perioperative mortality. PMID- 9346611 TI - Hepatitis C genotypes in liver transplant recipients: distribution and 1-year follow-up. AB - Chronic hepatitis C infection (CH-C) accounts for a significant number of patients undergoing orthotopic liver transplantation (OLT). Recently, hepatitis C virus (HCV) genotype-dependent differences in disease outcome and therapeutic responses have been suggested. The objectives of our study were to determine (1) the recurrence of HCV infection after OLT; (2) distribution of HCV genotypes in patients with CH-C who required liver transplantation compared with those who did not; and (3) the 1-year transplantation outcome in patients infected with different hepatitis C genotypes. RNA was extracted from sera of 20 patients who underwent OLT for end-stage liver disease secondary to CH-C (group I) and 52 patients with CH-C who did not require OLT (group II). For viral RNA detection, reverse transcriptase and polymerase chain reaction (RT/PCR) of 5'UT region was performed on all OLT patients both before and after OLT. For genotyping, RT-PCR of the NS 5 region was performed, followed by automated sequencing of the amplification products. Nineteen OLT patients had viral RNA detected by PCR both before and after OLT. One patient had no RNA detected before OLT but became viremic after OLT. The prevalence of HCV genotype 1b was significantly higher in group I patients compared with group II (53% v 23% respectively, P = .01). Examination of outcome at 1 year after OLT showed that 9 of 10 patients with HCV genotype 1b had histological evidence of hepatitis compared with 4 of 9 patients with other genotypes (non-1b) (P = .06). However, the number of patients who had one or more episodes of rejection, underwent retransplantation, or died at 1 year after OLT were similar. Recurrence of HCV infection after OLT was shown in all studied patients. Hepatitis C genotype 1b is more prevalent in our patients who underwent transplantation compared with a group with chronic hepatitis C who did not require transplantation (P = .01). Patients infected with HCV genotype 1b may have a higher risk of histological hepatitis after transplantation. PMID- 9346612 TI - Low incidence of intraspousal transmission of hepatitis C virus after liver transplantation. AB - Although the incidence of spousal transmission of hepatitis C virus (HCV) in chronic carriers is extremely low (1.4% to 8%), hepatitis C recurrence after liver transplantation is common with markedly increased serum HCV RNA levels. Thus, partners of these patients may be at higher risk of acquiring infection. This study evaluates the prevalence of spousal transmission of hepatitis C after liver transplantation. Twenty-two of 25 couples who were eligible agreed to the retrospective study. Twenty-two patients (17 males, 5 females) and spouses (5 males, 17 females) were studied with respective mean ages of 50.2 years (35 to 65 years) and 46.9 years (33 to 66 years). Liver enzymes, second-generation enzyme linked immunosorbent assay (ELISA) for antibody to HCV (anti-HCV) and HCV RNA by polymerase chain reaction (PCR), and branched DNA assay were performed. HCV associated antibodies were detected in 1 of 22 (5%) spouses and 21 of 22 (95%) patients (P < .0001). Nineteen of 22 (86%) patients tested positive by PCR with a mean value of 16,218,100 Eq/mL (464,700 to 51,980,000). All spouses including the only ELISA anti-HCV positive spouse tested negative by PCR (P < .0001). Eight of 21 spouses tested negative for anti-HCV pretransplantation, (13 of 21 pretransplantation were not tested). Estimated mean duration of hepatitis C infection in patients was 14 years (3 to 40 years). Mean patient follow-up posttransplantation was 654.5 days (141 to 1,959 days). Mean duration of marriage was 22.6 years (2.5 to 46 years). No risk factors other than exposure to index patients were observed in spouses. The incidence of spousal transmission of HCV in liver transplantation remains low (5%) and similar to chronic carriers of HCV. PMID- 9346614 TI - Progressive multifocal leukoencephalopathy after orthotopic liver transplantation. AB - Six weeks after liver transplantation, a 51-year-old man developed a slowly progressive hemiparesis with deteriorating mental status and seizures. Successive computed tomography (CT) scans of the brain revealed unilateral nonenhancing white matter lucencies that gradually coalesced and progressed to both hemispheres. Brain biopsy results were consistent with progressive multifocal leukoencephalopathy (PML). We believe this is the first antemortem description of PML after liver transplantation. Herein, we describe the case and review the literature on PML after solid organ transplantation. Early recognition of this central nervous system disease may be important with new advances in therapy of this viral infection of the immunocompromised patient. PMID- 9346613 TI - FK506 in liver transplantation for chronic hepatitis B: in vitro studies on lymphocyte activation and virus replication. AB - Recurrence of hepatitis B virus (HBV) in the graft is the major problem for patients with chronic HBV infection undergoing liver transplantation, which could be potentiated by the immunosuppression. In the present study, we used lymphocytes and hepatocytes isolated from patients with chronic HBV infection to investigate in vitro the effects of FK506 with and without methylprednisolone (25 ng/mL) on mitogen-induced lymphocyte proliferation, T-cell activation marker expression, and HBV replication in human hepatocytes. Increasing concentrations of FK506 (0.1, 0.5, and 5 ng/mL) resulted in a dose-dependent inhibition of lymphocyte proliferation with a reduction of the stimulation index by 9.2%, 39.0%, and 55.1%, respectively, with no difference between 25 chronic HBV carriers and normal controls. Methylprednisolone alone had no effect but potentiated the inhibitory effect of all three FK506 concentrations, such that the stimulation index was decreased by 20%, 56.2%, and 65.7%, respectively. FK506 (0.5 ng/ mL) reduced both the percentage of interleukin-2 receptor expressing T cells and the cell surface density of this receptor by 7.1% and 8.7% (P < .01), whereas it only reduced the proportion of HLA-DR expressing T cells by 6.8%. FK506 did not change significantly the intracellular HBV DNA or the hepatic expression of hepatitis B surface antigen (HBsAg) in short-term culture of human hepatocytes, whereas methylprednisolone increased the percentage of HBsAg positive hepatocytes in all 5 patients with active viral replication. These results indicate that the effects of FK506 on T-cell activation in chronic HBV carriers are identical to normal subjects, resulting in marked suppression of T lymphocyte function but only modest reduction in the expression of cell surface activation markers. The drug showed no direct stimulatory effect on viral replication, suggesting that FK506 can be a useful, steroid-sparing immunosuppressive agent for liver graft recipients with chronic HBV infection. PMID- 9346615 TI - Failure of liver transplantation to diminish cardiac deposits of amylopectin and leukocyte inclusions in type IV glycogen storage disease. AB - Orthotopic liver transplantation has been used to treat glycogen storage disease type IV. Most long-term surviving patients who have undergone liver transplantation have been free of neuromuscular and cardiac morbidity, and regression of cardiac amylopectin infiltration has been reported after liver transplantation. Leukocyte inclusions in glycogen storage disease type IV have also been reported. We present the case of a child who underwent orthotopic liver transplantation for glycogen storage disease type IV. In contrast to previous reports, at autopsy 2 1/2 years after transplantation, there was massive amylopectin deposits in his heart. Further, peripheral leukocytes never showed loss of amylopectin inclusions after transplantation. Orthotopic liver transplantation for type IV glycogen storage disease may not, in all cases, result in improvement in other affected organs. Consideration of multiorgan transplantation appears warranted. PMID- 9346616 TI - Percutaneous venovenous bypass in orthotopic liver transplantation. AB - Since January 1994, we have used percutaneous placement of both the subclavian and femoral cannulae to establish access for venovenous bypass during orthotopic liver transplantation. Percutaneous subclavian and femoral cannulae were used in 36 patients of which 5 had portal decompression by placement of a cannula in inferior mesenteric vein percutaneously through the abdominal wall. Intraoperative placement of the subclavian cannula is facilitated by placing a subclavian central venous line before the abdominal incision. One patient underwent exploration for femoral vein bleeding early in our experience. Another patient sustained hypotension as a result of a kinked subclavian cannula. In 4 patients, early in this experience, we had difficulty placing the subclavian cannula and resorted to axillary vein cut-down. There were no episodes of deep venous thrombosis detected by routine postoperative duplex ultrasonography. Minimum and maximum flow rates were significantly better (P < .01), with percutaneously placed cannulae in comparison to a control group of patients who underwent transplantation in whom we used the standard venous cut-down approach with a #7 Gott shunt (2.14 and 3.17 L/min v 1.65 and 2.41 L/min, respectively). Percutaneous placement of cannulae for venovenous bypass during liver transplantation is quick, safe, and effective. We would advocate this technique as an alternative approach for patients in whom bypass is deemed necessary. PMID- 9346617 TI - Safety, tolerability, and pharmacokinetic actions of diltiazem in pediatric liver transplant recipients on cyclosporine. AB - Thirty-two children who had undergone liver transplantation were paired according to their posttransplantation duration, renal function, and diagnoses when possible and randomized either to continue nifedipine (NIF group) or switch to diltiazem (DIL group), in addition to continuing their usual immunosuppressive medications. The cases were followed prospectively regarding diltiazem tolerance, cyclosporine dose requirements, effect on cyclosporine kinetics, diltiazem kinetics, as well as effect on renal function. Diltiazem was well tolerated at a dose of 3 mg to 6.4 mg/kg/day (max 180 mg/day) with infrequent self-limited mild side effects. Cyclosporine daily dose was reduced by a mean of 36.7% and 38.3% at 3 and 6 months, respectively, in the DIL group to achieve target trough cyclosporine levels without modifying liver function. No significant difference in renal function was observed after 3 to 6 months in either group based on blood urea nitrogen and creatinine levels and glomerular filtration rate by the DTPA method. Diltiazem appears to be well tolerated in children and allows for substantial dose reductions of CSA without apparent effects on liver graft function. PMID- 9346618 TI - Immunogenic role of Kupffer cells in a rat model of acute liver allograft rejection. AB - Kupffer cells (KCs) are of bone-marrow origin. After liver transplantation, recipient KCs are supposed to replace donor KCs. On the other hand, KCs are currently hypothesized to play a major immunogenic role in acute liver allograft rejection. In the present study, we investigated the immunogenic role of KCs in acute rat liver allograft rejection. For this purpose, we depleted the donor KCs using intravenous injection of liposome-encapsulated dichloromethylene diphosphonate (DMDP) in the fully allogenic ACI-to-LEW rat liver transplantation model. Kupffer cell depletion was confirmed using monoclonal antibodies ED2. In a first set of experiments, graft survival was evaluated, as were body weight and serum bilirubin changes, after the transplantation. Graft survival time showed no difference between the groups (treated, 12.5 +/- 0.92 days; control, 11.9 +/- 0.80 days). Body weight and serum bilirubin changes were similarly affected in both groups. In a second set of experiments, recipients were killed on day 6 after the transplantation, and rejection was histologically graded from 0 to 4. All grafted livers were judged as grade 3 regardless of treatment. ED2 staining showed KCs repopulation in both untreated and the dichloromethylene diphosphonate treated livers. The results of the present study provide evidence that KCs do not play an important immunogenic role in acute liver allograft rejection of the rat. PMID- 9346620 TI - More questions than answers. PMID- 9346619 TI - Quantitation of cytomegalovirus in the blood of liver transplant recipients. AB - An assay for quantitation of cytomegalovirus (CMV) has been developed. The assay combines DNA amplification and enzyme-linked immunosorbent assay (ELISA) detection. In this study, the assay has been used to examine sequential buffy coats from 32 consecutive liver transplant recipients. In a febrile patient, CMV titres in excess of 10(4) copies per 150,000 cells strongly suggest a diagnosis of symptomatic CMV infection. Antiviral therapy causes a rapid decline in viral titre. Viral titres are seen to rise presymptomatically in some patients. Median peak viral titres differ significantly between symptomatic patients (1.1 x 10(5)), asymptomatic CMV IgM-positive patients (1.7 x 10(3)), and asymptomatic CMV immunoglobulin (Ig)M-negative patients (2.9 x 10(2)). CMV quantitation can be used for diagnosis and surveillance and can also be used to monitor antiviral treatment. PMID- 9346621 TI - How valid is emergency liver transplantation for acute liver necrosis in patients with multiple-organ failure? AB - Multiple-organ failure (MOF), defined as the failure of initially uninvolved organs, is the final step of definitive and massive liver necrosis. Emergency liver transplantation (ELT) has radically modified the outcome of acute liver failure and early primary graft failure, but the results of ELT in cases of MOF are unknown. From May 1988 to June 1993, 243 patients underwent a liver transplantation (LT). Thirty-seven patients (15.2%) who had an acute liver necrosis complicated by a MOF underwent an ELT. Twenty-one patients were children. An emergency retransplantation was performed in 16 patients. Three or 4 organ-system failures (OSF) were present in 13 patients. Before ELT, the mean Acute Physiology and Chronic Health Evaluation (APACHE) II score was 26.3 +/- 5.1. Six-month and 1-year survival rates were 37.8% and 25.9%, respectively, after ELT complicated by MOF, and 78% and 73.5%, respectively, in other cases of LT. Twenty-six patients had surgical complications (70%), whereas thirty-one patients had medical complications (84%). Twenty-two patients died during the postoperative period (60%). Before ELT, infection (P < .05), cardiovascular failure (P < .03), and more than two OSF (P < .05) were more frequent in patients who died after intervention. The APACHE II score (P < .05) and the length of stay in the intensive care unit before ELT (P < .05) were lower among survivors. In the context of liver allograft shortage, our results suggest that an ELT should not be performed in patients with cardiac failure, more than two OSF, or an APACHE II score higher than 30. PMID- 9346622 TI - Prospective study comparing the efficacy of prophylactic parenteral antimicrobials, with or without enteral decontamination, in patients with acute liver failure. AB - The efficacy of prophylactic parenteral antibacterials, with or without selective decontamination of the digestive tract, was compared in patients with acute liver failure (ALF) or severe acetaminophen hepatotoxicity. One hundred eight patients were randomized on admission to receive intravenous ceftazidime and flucloxacillin, plus either oral and enteral decontamination with colistin, tobramycin, and amphotericin B (group 1), or enteral amphotericin B alone (group 2). The two groups were comparable with respect to age, gender, etiology, coma grade on admission, international normalization ratio, presence of renal failure, Acute Physiology and Chronic Health Evaluation II score, and indicators of poor prognosis. Patients were monitored for clinical and microbiological evidence of infection. There were 15 episodes of infection in 10 of 47 patients (21%) in group 1 and 17 episodes in 12 of 61 patients (20%) in group 2. No differences in incidence, site, and causative organisms of infection were observed between the two groups. Overall, the incidence of infection was significantly higher in patients who developed encephalopathy than in those who did not. In patients who on arrival were not encephalopathic, the development of infection was associated with progression to coma. Duration of Liver Intensive Care Unit (LICU) stay was an independent risk factor for the development of infection. Parenteral antibiotics are effective at reducing the risk of infection in patients with ALF; enteral decontamination provided no additional benefit. PMID- 9346623 TI - Central pontine myelinolysis with stupor alone after orthotopic liver transplantation. AB - Central pontine myelinolysis (CPM) after orthotopic liver transplantation is frequently diagnosed at autopsy. Its actual prevalence is unknown. We reviewed 386 orthotopic liver transplantations performed at the Mayo Clinic, Rochester, MN, including 39 with autopsy reports. CPM developed in four patients (1%) (found using magnetic resonance imaging in two, at autopsy in two). All four patients lacked typical signs and symptoms of CPM (pseudobulbar palsy and quadriplegia) but had stupor as the main presenting feature. When perioperative plasma osmolality and serum sodium values were compared with those values in patients with transient metabolic encephalopathy but no CPM at autopsy, no consistent association was observed, and values differed greatly. We conclude that CPM in orthotopic liver transplantation frequently is manifested by altered levels of consciousness alone, mimicking metabolic encephalopathy. PMID- 9346624 TI - Liver transplantation for adult polycystic liver disease. AB - Patients with adult polycystic liver disease and massive cystic replacement of the liver may present with severe debilitation and impairment of functional performance or, rarely, with signs of portal hypertension or hepatic dysfunction. In those patients incapacitated by severe hepatomegaly secondary to massive cystic replacement with predominantly small cysts (2 cm) without areas of parenchymal sparing, liver transplantation is a therapeutic option. Five patients with incapacitating symptoms from polycystic liver disease underwent liver transplantation as a final therapeutic procedure. Two patients had previous fenestration procedures without significant relief. All patients had radiographic evidence of concomitant polycystic kidney disease; two of these patients were dialysis-dependent at the time of liver transplantation. One patient underwent combined liver-kidney transplantation, whereas another received a six-antigen matched kidney transplant 64 months after liver transplantation. Four of five patients are alive 84, 39, 20, and 8 months after successful liver transplantation. All four have returned to normal functional status with complete resolution of symptoms. Liver transplantation is a suitable option for the patient with bilobar small cystic liver disease without areas of parenchymal sparing. However, only patients with severely compromised functional status should be offered this therapy. Concomitant renal evaluation is mandatory, and a knowledge of the natural history of this disease will aid in the decision of whether a combined liver-kidney transplantation is indicated. PMID- 9346625 TI - The influence of prostaglandin E1 on platelet adherence and injury in preserved rat liver allografts. AB - We have previously shown that part of the injury sustained by cold-preserved livers on reperfusion is the consequence of platelet adhesion to sinusoidal endothelium. The purpose of the present study was to determine whether prostaglandin E1 (PGE1) can reduce the injury and if so, how to maximize this beneficial effect. Rat livers were cold-preserved in University of Wisconsin solution for 30 hours then subjected to 1-hour warm ischemia after which they were reperfused at 37 degrees C with oxygenated Krebs-Henseleit solution with or without isolated platelets. PGE1 was used to treat the donor liver during harvesting, cold preservation, and reperfusion. In some studies, PGE1 was used to pretreat platelets before exposing them to the liver, and in other studies, both liver and platelets were treated. Pretreatment of platelets with paraformaldehyde, which inactivates them, or ADP, which activates them, was also studied. Treatment of livers with PGE1 significantly decreased preservation injury when livers were reperfused in the absence of platelets. However, when platelets were added to the perfusate, prior treatment of the liver with PGE1 had relatively minor beneficial effects. Pretreatment of platelets alone with PGE1 was also beneficial, but again the effect was small. However, when both liver and platelets were treated with PGE1 there was a highly significant decrease in the extent of liver injury and platelet adhesion. Perfusate transaminase levels were lower, bile flow was improved, and histologically, livers appeared less injured. Pretreatment of platelets with paraformaldehyde produced similar results to pretreatment with PGE1. When platelets were preactivated with adenosine diphosphate, extensive hepatic injury occurred upon reperfusion despite PGE1 treatment of the liver. PGE1 can lessen preservation-reperfusion injury impressively when administered to both liver and platelets but has little effect when platelets have been preactivated. PMID- 9346626 TI - Hepatic allograft rejection: regulation of the immunogenicity of human intrahepatic biliary epithelial cells. AB - Intrahepatic biliary epithelial cells were immunomagnetically purified from specimens of disaggregated human liver and were propagated in vitro. After three passes in culture, the cells were shown to be over 85% pure with contaminating leukocytes and endothelial cells constituting less than 2% of the population. Sensitive flow microfluorimetric analysis was performed after immunofluorescence labeling to quantify expression of both class 1 and class II major histocompatibility (MHC) antigens and the adhesion molecules intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and lymphocyte function-associated antigen-3 (LFA-3). It was shown that resting human intrahepatic biliary epithelial cells (HIBEC) expressed class 1 MHC antigens, ICAM-1, and relatively low levels of LFA-3. Stimulation with tumour necrosis factor-alpha (TNF-alpha) upregulated expression of ICAM-1, whereas Interferon gamma (IFN-gamma) and a combination of IFN-gamma and TNF-alpha upregulated class I MHC antigens and ICAM-1 and induced class II MHC molecules. The level of expression of MHC antigens and of ICAM-1 and LFA-3 after stimulation of HIBEC with combined IFN-gamma and TNF-alpha was comparable with the expression of these antigens by an Epstein-Barr virus-transformed B-cell line. The phenotypic similarity between cytokine-stimulated HIBEC and a known antigen presenting cell is consistent with a potential role for biliary epithelial cells in antigen presentation. PMID- 9346627 TI - Monoclonal antibodies against eosinophils in liver allograft rejection. AB - There has been recent interest in eosinophils as a histological diagnostic marker of liver allograft rejection. However, the reliability of counting eosinophils in sections stained with hematoxylin and eosin (H&E) has not been evaluated previously. We quantified eosinophils in 10 day-5 protocol liver biopsy specimens in 10 patients. The control groups were 5 patients with cytomegalovirus infection, 5 patients with obscure liver dysfunction, and 6 patients with HCV infection. Eosinophil count was assessed using H&E and by specific monoclonal antibody staining using (1) an anti-ECP antibody (EG2) and (2) a monoclonal antibody against human eosinophil major basic protein (MBP) (BMK-13). The average percentage of the total inflammatory infiltrate of eosinophils in portal tracts was 9% in the moderate to severe rejection group as compared with 0.25% in the mild rejection group (P < .001) and 0% in the control group (P < .001). The eosinophil count decreased markedly after successful treatment of rejection. The H&E staining correlated with MBP+ the (BMK-13 immunoreactive) cells but were more numerous with BMK-13. BMK-13 also stained significantly more cells when compared with EG2 (P < .01). This difference may be because EG2 staining only activated eosinophils, whereas BMK-13 is a pan-eosinophilic maker, regardless of activation. This study confirms that eosinophils are a specific feature of acute cellular rejection and are an aid to its diagnosis. BMK-13 is a useful pan eosinophilic marker that is more efficient in obtaining eosinophil count when compared with H&E. PMID- 9346628 TI - Posttransplantation de novo tumors in liver allograft recipients. AB - De novo cancers occurred after transplantation in 8,008 organ allograft recipients, who developed 8,531 different types of malignancy. Three hundred twenty-four liver recipients developed 329 cancers. There were striking differences in the patterns of neoplasms observed when these were compared with 7,200 tumors that occurred in renal allograft recipients. Lymphomas were much more common in liver allograft recipients (57% v 12% of all tumors), whereas skin cancers (39% v 15%), carcinomas of the cervix (4% v 1%), renal tumors (4% v 1%), and vulvar carcinomas (3% v 0.6%) were more common in renal allograft recipients. The high incidence of lymphomas is related partly to the more intense immunosuppressive therapy administered to hepatic allograft recipients and partly to the large percentage of pediatric patients among them. The intense immunosuppression also accounts for the much shorter induction times of lymphomas (mean, 15 v 46 months; P < .001) and nonlymphomatous tumors (mean, 27 v 72; P < .001) in liver compared with kidney recipients. The longer follow-up of renal recipients probably accounts for the higher incidence of the other tumors that tend to appear relatively late after transplantation. A remarkable feature was the high incidence of allograft involvement by lymphoma (44%). Complete remissions after treatment occurred in 11 of 28 patients in whom the lymphoma was confined to the allograft. A few tumors in liver recipients were related to the underlying disease for which transplantation was performed: hepatomas in patients who underwent transplantation for hepatitis B cirrhosis and colon carcinomas or cholangiocarcinomas in patients who underwent transplantation for chronic ulcerative colitis with sclerosing cholangitis. A surprising finding was the development of four leiomyosarcomas, three occurring in the allograft itself, in pediatric liver recipients. PMID- 9346629 TI - Fulminant hepatitis in patients undergoing liver transplantation: evidence for a non-A, non-B, non-C, non-D, and non-E syndrome. AB - Fulminant hepatic failure (FHF) in the absence of serum markers of hepatitis A (HAV) or B (HBV) infection or another cause is called non-A, non-B (NANB) FHF. The pathogenetic role of viral infection in NANB FHF remains controversial. To better define this relationship, we studied patients who underwent orthotopic liver transplantation (OLT) for FHF. Thirty-six patients with FHF underwent transplantation between 1987 and 1992. Pre-OLT serum was available for 24 patients, 14 with NANB FHF (all female; mean age, 32 years), and 10 (3 males, 7 females; mean age, 20 years) with a defined origin for FHF who formed the control group. Sera were tested using polymerase chain reaction for HAV, HCV, HDV, and HEV RNA and HBV DNA, and also serologically for antibodies to these viruses. In the NANB group, pre-OLT serum was negative for all viruses tested. Four patients in the control group had HBV serologically, 2 with HBV DNA in serum. One of these 4 also had hepatitis C and one hepatitis D infection. There was no difference in intensive care unit or hospital stay between the groups. The only significant difference in laboratory data was for peak creatinine pre-OLT (0.94 mg/dL in NANB v 1.62 mg/dL; P < .05). Two patients in the NANB groups and 3 in the control group required early retransplantation for graft primary nonfunction. One case of NANB FHF appeared to recur at 6 months but not after subsequent retransplantation. NANB FHF is not associated with hepatitis A, B, C, D, or E in plasma. It has a later age of onset but a similar clinical course to other forms of FHF and appears to preferentially affect women. PMID- 9346630 TI - Hepatitis C dilemma? PMID- 9346631 TI - Forum on critical care issues in liver transplantation: fluids, electrolytes, and renal disease in the perioperative liver transplant recipient. PMID- 9346632 TI - Surgical and anesthetic management of patients undergoing major hepatectomy using total vascular exclusion. AB - Total vascular exclusion (TVE) of the liver is accomplished by complete occlusion of inflow and outflow of the liver during hepatectomy. It affords the opportunity for bloodless, anatomically precise parenchymal transection but has not been widely used in this country. TVE should make it possible to treat large or unfavorably located lesions safely. To evaluate the benefit of this modality, we have examined the results of TVE in 49 major resections. Forty-nine patients with liver tumors (mean age, 50 +/- 17 years; range 3 to 75 years) were treated by the authors over 5 years with a mean age of 50 +/- 17 years (range 3-75). Thirty-five (71%) patients were females and 38 (78%) had malignant tumors (hepatocellular CA n = 15, liver metastases n = 20, other n = 3), whereas 11 (22%) had benign tumors (hemangiomas n = 7 other n = 4). Six (12%) had histological cirrhosis but normal liver function test results. Twenty two (45%) had previous surgery. Forty-seven (96%) underwent total or extended lobectomies. Two patients had segmental resection of benign tumors (one in segment 4 and one in segment 8). Mean surgical time was 4.7 hours (2.5-8.3 hours) and mean red blood cell requirement was 2.2 U (0 to 11). Twenty-two (45%) procedures were performed without transfusions. Hospital mortality rates were 0%. The mean postoperative hospital duration was 11 days (5 to 41 years). Complications occurred in 18 (36%), requiring reoperation in 1 case for wound debridement and in another for lysis of postoperative adhesions. Hepatic insufficiency occurred transiently in 2 patients with prolongation of protime and cholestasis and resolved within 4 days in 1 patient and 10 days in the other (with cirrhosis). The perception of hepatic resection as a prohibitive undertaking with high mortality rate may limit the use of resection in patients who might benefit from this modality. Our data document the effectiveness and safety of major hepatectomy even in cirrhotic patients using TVE. Expanded use of TVE and other advances in liver surgery should be considered to decrease the morbidity rate of resection and make the benefits of this therapy more widely available. PMID- 9346633 TI - The safety of continuous hepatic inflow occlusion during major liver resection. AB - To evaluate the safety of temporary hepatic inflow occlusion during major liver resection, we reviewed 71 consecutive noncirrhotic patients who underwent elective liver resection using this technique. There were 27 males and 44 females (mean age, 54.4 years), the majority of whom had hepatic malignancies. There were 31 right hepatectomies, 21 left hepatectomies, and 19 extended right hepatectomies. Ischemic injury of the liver was assessed using changes in postoperative liver function tests and patient outcome was assessed using morbidity and mortality rates. After preliminary ligation of the blood supply to the lobe to be removed, global hepatic ischemia was produced by temporary occlusion of the main portal vein and hepatic artery proper while the liver parenchyma was divided. The average duration of inflow occlusion was 59 minutes (range, 25 to 90 minutes). There was no operative mortality, and no patient developed liver failure. The liver enzymes reached their peak on the first postoperative day (mean aspartate aminotransferase [AST] level, 283 +/- 227 IU/L; mean alanine aminotransferase [ALT] level, 269 +/- 238 IU/L) and they returned to normal by 7 days. The most common postoperative complications were related to the chest, wound, and urinary tract. The mean intraoperative transfusion was 3.4 +/- 2.6 U of packed red blood cells, and 0.94 +/- 2.13 U of fresh frozen plasma. We conclude that continuous hepatic inflow occlusion for periods of 1 hour during major liver resection is safe and well-tolerated when there is no underlying parenchymal liver disease. PMID- 9346634 TI - Liver grafts can be preserved overnight. AB - The introduction of University of Wisconsin solution has made liver transplantation a semi-elective procedure. However, many studies have suggested that cold storage must not exceed 12 hours to avoid ischemic-type biliary complications, to reduce the incidence of primary nonfunction and to improve graft and patient survival. The aim of this study was to compare the function of livers transplanted as soon as possible after the liver was harvested and those preserved overnight. Over a 42-month period, we studied 133 elective orthotopic liver transplantation procedures. When cold ischemia started after 6 PM, patients underwent transplantation the following morning (group A), whereas the remainder underwent transplantation immediately (group B). Cold ischemia lasted 13.7 hours and 9.5 hours in groups A and B, respectively (P < .001). The two groups were comparable in terms of initial and late biochemical liver function, the rates of primary nonfunction (6.5% in group A, 6.8% in group B), acute rejection (45.6% in group A, 45.7% in group B), and vascular and infectious complications. No ischemic-type biliary complications were observed. Graft and patient survival were similar in both groups (72.4% v 75.4% and 72.9% v 75.8% in groups A and B, respectively). These results suggest that having taken a cut off at 6 PM to divide the groups into those that underwent transplantation consecutively and those deferred to the morning, the difference between the two groups in terms of storage is relatively modest. Elective liver transplantation can be performed after overnight graft storage without increasing short-term or long-term morbidity or mortality rates. PMID- 9346635 TI - Changes in liver core temperature during preservation and rewarming in human and porcine liver allografts. AB - Liver core temperature during organ procurement, storage, and rewarming has not been reported in human orthotopic liver transplantations (OLT). We have shown in the rat that optimal temperature for liver storage is not 4 degrees C but 0 degree C to 1 degree C. Therefore, a study was undertaken in humans and in pigs to determine the pattern of temperature change during OLT. The porcine studies were performed, because it was not possible to follow human grafts during the period that they were sterilely packaged. Temperature depression in humans was rapid during organ perfusion, remained stable during organ dissection, and decreased again slightly, when after excision, the organ was perfused again. Temperature depression during the period of perfusion with University of Wisconsin (UW) solution was curvilinear with the initial rapid temperature depression followed by a period of slower temperature depression. Volume perfused versus time was linear during these periods and the relationship between temperature depression and volume infused was curvilinear. At the time of packaging, 65 +/- 12 minutes after start of cold perfusion, the liver core temperature was 5.7 degrees C +/- 1.3 degrees C. Studies in the pig showed that it took 75 to 100 minutes for liver core temperature to decrease below 5 degrees C, and core temperature reached a plateau at 1 degree C at 195 +/- 75 minutes after packaging. During the rewarming period in humans, while vascular anastomoses were being constructed, there was a rapid linear increase in temperature from 0.8 degree C, when the graft was removed from the cold, to 17.2 degrees C +/- 3.1 degrees C at 45.5 +/- 4.4 minutes later, just before portal reperfusion commenced. These studies show that it takes only a short time to cool livers down to 10 degrees C, but after flushing is stopped, temperature depression is markedly reduced, and ideal temperatures are not reached before packaging. Rewarming of livers during performance of vascular anastomoses is rapid and reaches temperatures at which substantial hepatic metabolism is occurring. PMID- 9346636 TI - Superselective arterial embolization in the liver transplant recipient: a safe treatment for hemobilia caused by percutaneous transhepatic biliary drainage. AB - Transcatheter arterial embolization is widely used to treat life-threatening iatrogenic hemobilia, although in the transplanted liver its use has only been reported in two cases. To evaluate more fully whether transcatheter embolization is safe and effective in transplant recipients, we retrospectively reviewed eight cases of severe hemobilia. These occurred after 128 percutaneous transhepatic biliary drainage procedures performed during a 6-year period. In each case, angiography localized the bleeding to a specific intrahepatic branch that was then subselectively catheterized and occluded by transcatheter embolization. Bleeding was successfully controlled by this method in all eight patients with no immediate complications. The main, right, and left hepatic arteries were shown to be patent immediately after embolization by angiography in all patients. Duplex sonography performed in each case (1 to 4 months) after the procedure confirmed that patency was maintained in all patients. No patients developed liver abscesses, sepsis, or clinical liver infarctions after the embolization. No patients underwent retransplantation after embolization. Our experience shows that superselective transcatheter embolization is a safe, effective therapy to correct iatrogenic hemobilia in the liver transplant recipient without threatening the patency of the major hepatic arteries, the viability of the liver, or the integrity of the biliary tree. PMID- 9346637 TI - Hepatitis C virus in body fluids after liver transplantation. AB - Recurrence of hepatitis C virus (HCV) after liver transplantation is common and is associated with high blood levels of HCV RNA. Higher blood levels of HCV may promote body fluid expression of the virus. We tested 152 body fluid specimens from 33 patients with chronic hepatitis C, 21 of whom had undergone prior liver transplantation. All patients had hepatitis C viremia, as determined by a reverse transcription polymerase chain reaction (PCR) to the 5' noncoding region. The virus was quantitated in serum by the branched chain DNA assay (bDNA). Body fluids (33 sputum, 33 saliva, 33 urine, 32 tear, 9 vaginal, and 12 semen samples) were analyzed using PCR for HCV RNA. Serum HCV RNA by bDNA in the posttransplantation group was 255 +/- 229 x 10(5) compared with 50 +/- 56 x 10(5) eq/mL in the patients who did not undergo transplantation (P = .01). All urine, tear, and semen specimens were negative for HCV RNA. Five of 21 (24%) posttransplantation patients had detectable HCV RNA using PCR in oral secretions compared with 0 of 12 patients who did not undergo transplantation (not statistically significant). However, 5 of 11 patients with serum HCV RNA by bDNA results greater than 150 x 10(5) eq/mL had positive RNA in oral secretions compared to 0 of 22 patients with bDNA less than 150 x 10(5) eq/mL (P = .01). Posttransplantation patients were more likely to have bDNA levels exceeding 150 x 10(5) eq/mL (11 of 21 v 1 of 12, P = .03). Patients within the first year of transplantation were particularly prone to viral RNA levels exceeding 150 x 10(5) eq/mL (8 of 9 v 3 of 12, P = .01). We conclude that HCV RNA can occasionally be detected using PCR in oral secretions after liver transplantation. This is more likely during the first year posttransplantation when blood levels of HCV RNA often exceed 150 x 10(5) eq/mL by the branched chain DNA assay. Whether or not these observations represent an increased risk of transmission of infection during the early posttransplantation period is not certain. PMID- 9346638 TI - Hepatitis B transmission from a liver donor who tested negative for hepatitis B surface antigen and positive for hepatitis B core antibody. PMID- 9346639 TI - Interferon alfa for recurrent hepatitis B infection after liver transplantation. AB - Reinfection with hepatitis B virus (HBV) after liver transplantation is nearly universal in patients not receiving immunoprophylaxis. Because reinfection reduces graft and patient survival, treatment of recurrent infection is important. Interferon alfa (IFN-alpha) is an effective therapy for chronic hepatitis B infection in immunocompetent patients, but its efficacy in transplant recipients has not been established. Fourteen liver transplant recipients with recurrent hepatitis B infection (hepatitis B surface antigen [HBsAg] positive in serum; hepatitis on biopsy) were treated with IFN-alpha 2b (Intron A; Schering Inc, Kenilworth, NJ) 3 million units (MU) three times weekly for 23.5 weeks (median, range 4 to 41). The primary endpoint was loss of HBV DNA by the b-DNA assay (a virological response). Before treatment, all patients were HBV DNA positive and 9 were hepatitis B e antigen (HBeAg) positive. Pretreatment HBV DNA levels were 6,760 MEq/mL (median, range 2.0 to 11,888 MEq/mL). HBV DNA levels decreased significantly with treatment (P = .03). Four patients had a complete and sustained virological response. Virological responses did not consistently correlate with biochemical response because of concomitant hepatitis C. Two patients had a serological response; 1 lost HBeAg, another lost HBeAg and HBsAg. All responders remained HBV DNA negative in follow-up (mean, 32 months; range, 23 to 40), but 1 patient required retransplantation for cirrhosis. Of the nonresponders, 1 patient required retransplantation for chronic rejection, 3 required retransplantation for recurrent hepatitis B, 3 died with recurrent hepatitis B, and 3 are alive and remain HBV DNA positive. IFN-alpha can induce a sustained serological (14%) and virological response (29%) in liver transplant recipients with recurrent HBV infection. PMID- 9346640 TI - Transjugular intrahepatic portosystemic shunt in patients with end-stage liver disease: results in 85 patients. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is becoming an accepted procedure as a bridge to orthotopic liver transplantation (OLT) in patients with end-stage liver disease (ESLD) and bleeding from portal hypertension. It allows the immediate control of acute bleeding and decreases the risk of recurrent acute bleeding while the patient is awaiting OLT. We review in this report, our experience with 85 patients who underwent a TIPS procedure for gastrointestinal variceal bleeding from September 1991 until April 1994. All patients had liver cirrhosis and all had previous sclerotherapy before TIPS. Child-Pugh score was calculated at enrollment, and all patients were evaluated for possible OLT. Thirteen patients were Child A, 49 were Child B, and 23 were Child C. Fifty-three patients were candidates for OLT, and 32 were not. TIPS was performed urgently in 25 patients. At a median follow-up of 582 days (range, 1 to 1,095), 35 patients underwent transplantation, 21 patients died, and 29 patients are still alive and did not undergo transplantation. Technical complications were observed in 7% of patients and new onset of clinical encephalopathy in 37%. The 30-day mortality rate after TIPS was 13%. Actuarial survival was 60% at 1 and 3 years. Child class C and urgent TIPS were shown to be two independent predictor factors for mortality. TIPS was shown to be a valuable procedure, not only as a bridge to OLT but also as palliation for bleeding from portal hypertension in patients who were not candidates for either surgical shunt or OLT. However, its role in bleeding patients with acceptable liver function needs further investigation. PMID- 9346641 TI - Effect of orthotopic liver transplantation on employment and health status. AB - Employment, functional status, health status, and prevalence of anxiety and depression were assessed in patients who had undergone orthotopic liver transplantation at Duke University from 1984 to 1993 to identify social and economic factors that might influence return to work after liver transplantation. Patients were asked to complete mailed questionnaires. A transplant nurse coordinator assigned patients a Karnofsky score, unaware of the questionnaire responses. The response rate was 71% (52 of 72 patients). The median age of the post-liver transplantation patients was 49 years. Median years of education were 13. Sixty-five percent of patients were male. Sixty percent of patients were employed posttransplantation. Employed and unemployed posttransplantation patients showed no significant difference in age, education, gender, marital status, race, family coping skills, or cause of liver disease. Return to work after transplantation did not correlate with socioeconomic status or spouse's employment. Posttransplantation return to work was highly correlated with pretransplant employment (P < .0005). The prevalence of anxiety and depression, assessed by the Hospital Anxiety and Depression Scale (HAD), was 9% and was no different in the employed or unemployed patients. Health status, as measured by Karnofsky score, was excellent; all patients received Karnofsky scores > or = 80%. Health perceptions were compared in employed versus unemployed posttransplantation patients with the SF-36, a 36-item short form survey developed by the investigators of the Medical Outcome Study. This revealed significantly different values in the subscale, physical functioning, with a mean score of 70.6 in the employed and a mean score of 48.4 in the unemployed posttransplantation patients (P = .004) and role-physical with a mean score of 61.8 in the employed and a mean score of 27.6 in the unemployed posttransplantation patients (P = .005). Eighty percent of patients not returning to work cited "problems with their health" as their major obstacle to employment. Although objective health status was good to excellent in all patients after transplantation, patients perceived that their health status was poor, with the lowest scores observed in unemployed posttransplantation patients. PMID- 9346642 TI - The natural history of untreated focal allograft rejection in liver transplant recipients. AB - Focal rejection involves less than 20% of portal tracts in liver allograft biopsy results. The clinical significance of "focal" rejection on protocol liver biopsy results is unknown. The purpose of this study was to prospectively determine the incidence of clinically significant rejection in patients with focal rejection after orthotopic liver transplantation. Biopsy specimens from 165 consecutive transplantations in 149 patients were analyzed. After protocol biopsy specimens were obtained, patients with focal or mild rejection were observed. Fifty of 583 (8.6%) protocol biopsy results in 41 patients showed focal or mild rejection. None were treated on the basis of this histological finding. Six patients subsequently developed abnormal liver function tests and required treatment with additional immunosuppression. We conclude that focal or mild rejection is a relatively common finding on protocol liver biopsy results and only rarely progresses to a clinically significant problem. Patients with this finding can safely be observed without treatment. PMID- 9346643 TI - Transmission of hepatitis B virus through allotransplantation. PMID- 9346644 TI - Is hepatic histology the true gold standard in diagnosing acute hepatic allograft rejection? PMID- 9346645 TI - United Network for Organ Sharing center-specific data: our report card. PMID- 9346646 TI - Prostaglandins in liver failure and transplantation: regeneration, immunomodulation, and cytoprotection. Prostaglandins in Liver Transplantation Research Group. AB - Prostaglandins (PG) are involved in the regulation of many physiological processes in the liver and play a major role in the pathophysiology and treatment of liver diseases. In addition to their effects of cell growth and immune function, PGs have shown cytoprotective effects on hepatocytes in various toxic, ischemic, and infectious models of liver injury. Although the mechanisms for these beneficial effects have not been precisely delineated, synthetic PG analogues have increasingly been used in patients with acute liver failure and chronic liver disease. There is also increasing evidence suggesting that PGs may reduce the early morbidity and mortality associated with liver transplantation, particularly in the context of primary graft nonfunction and renal dysfunction associated with cyclosporine and tacrolimus therapy. PG analogues have also been used for the treatment and control of recurrent hepatitis B virus infection in liver allograft recipients. The purpose of this review is to evaluate the role of PGs in hepatic physiology and disease and to review the use of synthetic PG analogues in the clinical settings of liver failure and transplantation. PMID- 9346647 TI - Failed allografts and causes of death after orthotopic liver transplantation from 1985 to 1995: decreasing prevalence of irreversible hepatic allograft rejection. AB - Mayo Clinic pathology files from March 1985 to March 1995 contained records of 584 orthotopic liver transplantations in 515 patients. The most common indication for liver transplantation was primary sclerosing cholangitis (PSC), followed by primary biliary cirrhosis (PBC), and cryptogenic cirrhosis. In 59 patients, a total of 69 single or repeated retransplantations became necessary. Vascular complications necessitated retransplantation in 35 cases, followed by irreversible rejection in 16 cases, and primary graft failure in 8 cases. Ninety nine patients died, 25 of them after one or more retransplantations. Infectious complications caused death in 38 cases, followed by graft-related complications (excluding rejection) in 22 cases, noninfectious systemic diseases such as intracerebral hemorrhage (21 cases), malignancies in 13 cases, and irreversible rejection in 5 cases. In the decade from 1985 to 1995, only 14 patients had irreversible rejection that often was associated with other complications such as ischemic cholangitis. Furthermore, the rate of irreversible rejection decreased dramatically. Thus, although 5 deaths were caused by irreversible rejection, none occurred since March 1991, and of the 16 retransplantations for irreversible rejection, only one needed to be performed during these last 4 years. The disappearance of irreversible rejection, which can be described as the "vanishing vanishing bile duct syndrome," must be considered when new immunosuppressants are tested, because graft loss and death from rejection are no longer suitable criteria. PMID- 9346648 TI - Fibrous tumor-liver interface in large hepatic neoplasms: its significance for tumor resection and enucleation. AB - Based on the clinical observation that large tumors located in the liver may be resected or enucleated with relative ease, the present retrospective study aimed at the detailed assessment of the tumor/liver interface in large hepatic neoplasms. For this purpose, a systematic light microscopic and immunohistochemical study on resection specimens of 10 giant hemangiomas, 10 hepatocellular carcinomas, and 9 liver metastases was performed. In giant hemangiomas, four distinct interface patterns were identified: fibrous interface (pattern A), interdigitating interface (pattern B), compression interface (pattern C), and irregular/spongy interface (pattern D). In 5 of 10 of these tumors, a single interface type (pattern A) was observed, whereas the other five tumors exhibited a mixed interface type, although two of them had a predominance of pattern A. Overall, 7 of 10 giant hemangiomas were exclusively or partially separated from liver substance by a distinct fibrous interface. Similarly, a fibrous interface of variable thickness and containing remnants of preexisting portal tracts was observed in 18 of 19 malignant epithelial tumors, and in 10 of 18 tumors, this fibrous zone was complete, ie, totally covering the tumor border facing the resection surface. These findings indicate that large hepatic tumors, both benign and malignant, can induce a distinct fibrous interface in a large proportion of cases. We suggest that such an interface, grossly visible as a capsulelike structure, plays a significant role for the resectability of large liver tumors. PMID- 9346649 TI - Hepatitis C virus genotypes, hepatitis, and hepatitis C virus recurrence after liver transplantation. AB - Several genotypes of hepatitis C virus (HCV) have been recently identified by phylogenetic analysis, but their clinical relevance in the liver transplant setting is unknown. We evaluated the incidence and course of recurrent hepatitis C after transplantation in 50 patients who underwent transplantation for HCV related liver disease. Liver biopsy specimens were obtained when clinically indicated and at yearly intervals; hepatitis was histologically graded and staged according to standard criteria. HCV-RNA was detected by nested reverse transcription polymerase chain reaction (RT-PCR). HCV genotyping was performed by primer specific PCR. Follow-up was 6 to 62 months. HCV genotype distribution after transplantation of our 50 patients was as follows: 31 type 1b, 13 type 2a, 3 type 1a, 1 type 3a, 1 type 1b/2a, and 1, undetermined. Actuarial rates of recurrent hepatitis and of severe fibrosis or cirrhosis 5 years after transplantation were 56% and 20%, respectively, in patients infected by type 1b and 33% (P = .18) and 8% (P = .16) in those infected by 2a. In conclusion, this study provides evidence that in patients infected by HCV type 1b there is a trend for a more aggressive recurrent liver disease. PMID- 9346650 TI - Sanctuary of hepatitis B virus in bone-marrow cells of patients undergoing liver transplantation. AB - Hepatitis B virus (HBV) reinfection after liver transplantation is a major problem. HBV is mainly a hepatotrophic virus but replicates in many extrahepatic tissues. We present here two cases of infected patients who underwent liver transplantation. Both underwent bone marrow (BM) and liver biopsies after transplantation. Biopsy specimens were stained for hepatitis B surface antigen (HB-sAg), and bone marrow aspirates and were separated for all subsets of cells. In both cases, HBV DNA analysis detected DNA in all BM fractions after transplantation, but HBV recurrence was found only in one case. We suggest that graft reinfection after liver transplantation may be caused by active replication of HBV in extrahepatic tissues and that BM cells are probably one of the major sanctuaries. The use of immunoprophylaxis based on BM-HBV studies is discussed. PMID- 9346651 TI - The use of lidocaine as a test of liver function in liver transplantation. AB - The hepatic metabolism of lidocaine to monoethyl-glycinexylidide (MEGX) is the basis of a dynamic test of liver function. To understand its potential value in liver transplantation, the latter has been considered in the following three separate stages: pretransplantation assessment of potential candidates, potential liver donors, and the transplant recipient. In pretransplantation patients, data support its role in assessing risk of morbidity and mortality. In assessment of the liver transplant donor, there are differences concerning apparent usefulness, and these must be resolved. In the liver transplant recipient, serial measurements are useful to measure real-time hepatic metabolic activity. Low MEGX values reflect the clinical condition of the patient, and the importance of entirely assessing the patient, not just noting the test result, is paramount. This review has considered the role of the MEGX test in liver transplantation. PMID- 9346652 TI - Orthotopic liver transplantation for bone-marrow transplant-associated veno occlusive disease and graft-versus-host disease of the liver. AB - Bone marrow transplantation (BMT) is a highly successful and curative therapy for many primary hematologic malignancies. However, hepatic dysfunction and failure are important causes of morbidity and mortality in this patient group after BMT. Hepatic failure can occur as a result of involvement by graft-versus-host disease (GVHD) or as a result of veno-occlusive disease (VOD). Therapies for these complications are often ineffective, especially for VOD, as approximately one fourth of patients develop irreversible liver disease and die of multiorgan failure. Accordingly, we have reviewed our center's experience with orthotopic liver transplantation (OLT) to manage the hepatic complications of BMT. We describe two patients who were treated with OLT after developing hepatic failure post-BMT. One patient had VOD with mild cutaneous and gastrointestinal GVHD; in the other patient, the pathophysiologic process affecting the liver was severe GVHD. Based on our center's experience and review of the literature, we believe OLT should be considered in patients with severe hepatic dysfunction post-BMT. PMID- 9346653 TI - Regeneration of a transplanted liver after right hepatic lobectomy. AB - Liver regeneration has been described after heterotopic liver transplantation, small-for-size orthotopic liver transplantation and reduced-size liver transplantation. In this report, we document the regenerative response of a whole liver transplant to major resection for the first time. A right hepatic lobectomy for liver ischemia was performed in a 30-year-old female after transplantation for autoimmune disease of the liver. Volumetric analysis of computed tomography (CT) scans revealed a preoperative liver volume of 1,961 mL, whereas analysis of the 6-week posthepatectomy CT scan showed a volume of 1,820 mL. Factors influencing regeneration in the setting of a liver transplant include rejection, ischemia/thrombosis, infection, or cyclosporine hepatotrophic/hepatotoxic effects. These factors, balanced with the intrinsic ability of hepatocytes to achieve a standard liver volume, determine the extent of regeneration. PMID- 9346654 TI - Adult presentation of diffuse bile duct stenosis: therapy with liver transplantation. PMID- 9346655 TI - Advances in biliary reconstruction after liver transplantation. PMID- 9346656 TI - Prostaglandins for clinical use: are we there yet? PMID- 9346657 TI - The lidocaine monoethylglycinexylidide test of liver function. PMID- 9346658 TI - Famciclovir treatment of hepatitis B virus recurrence after liver transplantation: a pilot study. AB - Despite hepatitis B immunoprophylaxis hepatitis B virus (HBV) recurrence is a frequent and often fatal complication after orthotopic liver transplantation (OLT). The purine nucleoside analogues penciclovir and its oral form famciclovir (FCV) proved to be well tolerated and effective against herpes simplex and zoster virus infections. In addition, an effective reduction of duck and human HBV replication was observed. Therefore, we conducted an uncontrolled pilot study of famciclovir in patients with HBV recurrence after OLT. Twelve patients have received famciclovir for at least 3 months in an open compassionate-use protocol. FCV was administered orally 500 mg three times a day for all patients (except one patient who was started on 750 mg three times a day for the first 2 weeks). Immediately after starting famciclovir, serum HBV DNA levels declined in 9 of 12 patients (75%) with a mean reduction from baseline levels of 80% after 3 months, 90% after 6 months, and > 95% after 12 months of treatment. With continued treatment, 5 of these 9 patients became negative by conventional hybridization assay, and in one of these HBV DNA became undetectable by polymerase chain reaction (PCR) 28 weeks after the start of treatment. Three patients showed no (sustained) reduction in HBV DNA after at least 3 months of treatment; therefore, FCV was stopped. Latest serum alanine aminotransferase (ALT) levels decreased in 6 of 12 patients (50%) with a median decrease of 80% (range, 40%-95%) in comparison to pretreatment ALT values. ALT levels normalized in 4 patients (33%). One patient died due to sepsis and peritonitis in week 13 of treatment. This event was not related to FCV. No clinically significant side effects were noticed in any patient. The oral nucleoside analog famciclovir reduces HBV replication and transaminase levels in patients with HBV recurrence after liver transplantation. Because long-term FCV treatment is well tolerated, famciclovir appears to be a promising antiviral strategy in the treatment of HBV in immunocompromised patients. PMID- 9346659 TI - Beneficial effect of ribavirin on hepatitis C-associated cryoglobulinemia after liver transplantation. AB - Mixed cryoglobulinemia is a well-known complication after hepatitis C virus (HCV) infection. We report five cases in which cryoglobulinemia appeared or grossly exacerbated following orthotopic liver transplantation (OLT). Cryoglobulinemia and the associated clinical symptoms resolved or improved in two patients treated with ribavirin after liver transplantation, while plasmapheresis was ineffective in another patient. The mechanism involved in induction of cryoglobulinemia after liver transplantation is unknown. However, the effect of antiviral therapy observed in these patients suggests a correlation between cryoglobulinemia, HCV replication, and possibly hepatocellular disease activity. A larger-scale study is warranted to test the effect of ribavirin on post-OLT HCV-associated cryoglobulinemia. PMID- 9346660 TI - Follow-up after liver transplantation for protoporphyric liver disease. AB - Protoporphyria is a genetic disorder in which patients may develop severe protoporphyrin-induced liver damage and require transplantation. Because unique problems occur in the perioperative period and because excess production of protoporphyrin by the bone marrow continues after liver transplantation, the efficacy of this procedure for protoporphyric liver disease is uncertain. We present follow-up of nine patients who underwent liver transplantation. Two patients died within 2 months of transplantation, one from complications of abdominal bleeding and the other from sepsis after bowel perforations. The remaining seven patients had follow-up at 14 months to 8 years after transplantation (mean, 3.8 years). Two of the seven had suffered skin burns from exposure to operating room lights, which healed without scarring. Three had axonal neuropathies in the postoperative period requiring prolonged mechanical ventilation, and motor defects persisted in two. Five patients had normal liver chemistries at follow-up (mean, 3.5 years), with liver biopsy results normal or showing mild portal triad abnormalities, but erythrocyte protoporphyrin levels remained significantly elevated (1,765 +/- 365 mcg/dL; normal, < 65). The other two patients, both of whom had rejection, cytomegalovirus infection, and biliary tract obstruction requiring endoscopic therapy, had a recurrence of protoporphyric liver disease as indicated by liver biopsy features. One died 5 years after transplantation from complications of the liver disease. The other was stable 3.3 years after transplantation and was being monitored for possible retransplantation. Thus, liver transplantation can be performed successfully in patients with protoporphyric liver disease, with intermediate survival rates comparable to the general transplant population. However, disease may recur in the graft, particularly if there are complications that cause cholestasis. PMID- 9346661 TI - Morbidity in patients with posttransplant diabetes mellitus following orthotopic liver transplantation. AB - It is not well understood whether posttransplant diabetes mellitus (PTDM) following orthotopic liver transplantation (OLTx) alters postoperative morbidity. This study was designed to evaluate this question. All adult patients who received an OLTx between July 1985 and March 1993 (n = 497) were evaluated by retrospective chart review for evidence of PTDM after OLTx. The patients identified with PTDM (n = 26) were case matched with nondiabetic OLTx recipients based on primary liver disease diagnosis, age, gender, date of first OLTx, and survival. Liver synthetic function, number and severity of rejection episodes, graft survival, total number of hospital days within the first year post-OLTx, renal function, and number and type of infection episodes were analyzed to assess differences in morbidity between the PTDM and control patients after OLTx. Of the 497 adult patients who underwent OLTx, 26 (5.2%) were identified as having PTDM within 1 month of discharge. Factors which identified individuals at higher risk for DM after OLTx included higher pre-OLTx fasting blood glucose (P = .04); lower body mass index after OLTx (P = .02); and cyclosporine rather than OKT3 induction (P = .009). Graft survival, synthetic function, and the total number of rejection episodes during the first year were not different between the two groups. The morbidity variables of total number of days in the hospital during the first 12 months, renal function, and type and number of infections were also similar between the two groups. In summary, 5.2% of adult patients developed DM within 1 month of OLTx. Pre-existing insulin resistance, postoperative stress, and immunosuppression medications all likely contribute to the development of overt hyperglycemia after OLTx. Although PTDM can be a consequence of OLTx, it does not have a significant impact on patient outcome in the first year after OLTx. PMID- 9346662 TI - DNA image analysis study of lesions of the gallbladder and biliary system. AB - DNA ploidy analysis of 22 lesions arising from the intrahepatic and extrahepatic biliary systems and gallbladder was performed on Feulgen-stained 5-microns sections from archival paraffin-embedded tissue on an image analyzer (CAS-200). The cases included intrahepatic cholangiocarcinoma (n = 6), extrahepatic bile duct carcinoma (n = 5), carcinoma of the gallbladder (n = 11), and appropriate controls. All malignancies were stage III and IV adenocarcinomas with the exception of 1 stage II moderately differentiated gallbladder adenocarcinoma. No correlation with ploidy and stage could be made, most likely because of the advanced stage of the tumors at the time of presentation. When DNA ploidy was compared with the grade of tumor, the 3 well-differentiated adenocarcinomas (2 cholangiocarcinomas, 1 bile duct carcinoma) were predominantly diploid; however, diploid peaks were also found in moderately differentiated adenocarcinomas of all 3 sites and in a poorly differentiated adenocarcinoma of the extrahepatic bile ducts. Finally, 15 of 22 (68%) of cases showed only aneuploid populations. Multiple populations were observed in 19 of the 22 cases; this finding may reflect intratumoral heterogeneity and correlate with the advanced stage and aggressive nature of malignancies of the gallbladder and biliary system. PMID- 9346663 TI - The surgical challenge of papillary neoplasia of the biliary tract. AB - This study is a case report and literature review of the surgical approach to papillary lesions of the biliary tract exclusive of the ampulla of Vater. Papillary lesions of the bile ducts, exclusive of the ampulla of Vater, are distinctly uncommon but, because of their unpredictable and aggressive behavior, pose challenging problems for the surgeon. Including the present illustrative case description, an English language literature review was conducted to determine the number and clinical behavior of papillary lesions of the bile ducts, particularly the propensity for malignant transformation, and the most favorable surgical approach. In addition to the present case, 29 patients with papillary biliary lesions were found in the literature. Twenty-two patients had tumors in multiple locations in the biliary tract, 6 had isolated lesions in one hepatic duct, and 3 had diffuse papillomatosis. Overall, 15 of 30 patients (50%) died of their disease. Patients with solitary tumors faired best (5 of 6 long term survivors), and patients with diffuse papillomatosis did worse (all 3 died). Characteristics of these lesions include a propensity for recurrence (15 of 30 patients), an abundant mucin production, and a tendency toward malignant change (eight of 30 patients). Papillary lesions prove difficult to treat because of their frequent multifocality, propensity to recur after surgical extirpation, and malignant potential. Surgical strategy should include liver resection for isolated tumors, close monitoring for recurrence, and reoperations as required to control tumor growth. PMID- 9346664 TI - Toxoplasma gondii pneumonitis in a liver transplant recipient: implications for diagnosis. AB - Toxoplasma gondii pneumonia developed 27 days after retransplantation for chronic rejection in a liver transplant recipient receiving tacrolimus as primary immunosuppression. Trimethoprim-sulfamethoxazole had been discontinued (due to cytopenia) before the onset of T gondii pneumonia. T gondii was isolated from bronchoalveolar lavage fluid by growth in fibroblast cell culture routinely used for the isolation of viruses and detected in lung on autopsy. Tissue culture used for the detection of viruses can also be diagnostically useful for the detection of T gondii. PMID- 9346665 TI - Wound recurrence after resection of hepatocellular carcinoma. AB - Cutaneous metastases arising along the exit site of an abdominal drain in one patient and within the operative wound in two others after resection of a hepatocellular carcinoma (HCC) are reported. All patients underwent curative hepatic resection, and cutaneous metastases occurred 9, 12, and 22 months after surgery respectively. Cutaneous metastases were not associated with intrahepatic recurrence and were treated by local excision. Two patients are alive and disease free 48 and 49 months after hepatic resection, respectively; the third patient died from recurrence 39 months later. These observations suggest that cutaneous malignant seeding may occur, and the authors recommend the observance of special care during liver surgery to prevent occurrence of this complication, including the use of wound protection and avoiding the opening of the specimen by the surgeon. The follow-up of patients who have undergone liver resection for an HCC should include the examination of all operative wounds including the exit site of abdominal drains. These subcutaneous nodules should be resected as an appreciable survival can be expected. PMID- 9346666 TI - Expanding the immunosuppressive repertoire. PMID- 9346667 TI - Pathophysiology of arterial hypoxemia in advanced liver disease. PMID- 9346668 TI - Scenario number one: hepatopulmonary syndrome. AB - HPS is an increasingly recognized clinical entity resulting from intrapulmonary vasodilatation in patients with liver disease and/or portal hypertension. The pathogenesis of alterations in the pulmonary vascular bed in affected patients is poorly understood and the subject of ongoing investigation. The differential diagnosis of pulmonary symptoms and gas-exchange abnormalities in patients with liver disease being evaluated for transplantation is broad and should be focused on differentiating pulmonary causes that significantly increase the risk for transplantation from HPS where transplantation has emerged as a useful treatment. Specific contraindications to transplantation in patients with HPS have not been identified, though unique postoperative complications have been observed and may be treatable during the frequently prolonged resolution of intrapulmonary vasodilatation. The development of a database of information on patients with HPS undergoing transplantation will provide insight into potential contraindications, prognostic features, and postoperative complications unique to these patients. PMID- 9346669 TI - Scenario number two: pulmonary hypertension. PMID- 9346670 TI - Scenario number three: cardiopulmonary management. PMID- 9346671 TI - Shortage of organs with a consequent increase in stand-by time and patient deaths. PMID- 9346672 TI - Biliary reconstruction and biliary complications in liver transplantation. PMID- 9346673 TI - Increased expression of proliferating cell nuclear antigen in liver allograft rejection. AB - Detection of proliferating cell nuclear antigen is useful for the study of proliferative activity of neoplastic and non-neoplastic lymphoid, parenchymal, and mesenchymal cells. Allograft rejection is associated with the recruitment of circulating cells and their proliferation in the graft. The intrahepatic expression of proliferating cell nuclear antigen on paraffin-embedded liver biopsy specimens (n = 110 from 32 patients) was examined by an avidin-biotin peroxidase method using a monoclonal antibody to proliferating cell nuclear antigen. The percentage of positive nuclei was determined in hepatocytes, biliary epithelium, and lymphocytes. There were four histologic groups: 1, moderate-to severe rejection (n = 19); 2, mild rejection (n = 28); 3, nonspecific inflammation (n = 45); and 4, donor livers (n = 18). The percentage of positive nuclei was higher in group 1 compared to group 2 (hepatocytes p = 0.01; biliary epithelium p = 0.0007; lymphocytes p = 0.0001), to group 3 (hepatocytes p = 0.0002; biliary epithelium p = 0.0001; lymphocytes p = 0.0001), and to group 4 (for all three locations p < 0.0001). When group 2 was compared to group 3 the results were significant for biliary epithelium (p = 0.0001) and lymphocytes (p = 0.0001), but not for hepatocytes (p = 0.07). We conclude that proliferating cell nuclear antigen expression, especially in lymphocytes, correlates with the severity of histologic rejection. Proliferating cell nuclear antigen expression may be useful in predicting the progression and response to different antirejection therapies. PMID- 9346674 TI - Ionized hypomagnesemia in patients undergoing orthotopic liver transplantation: a complication of citrate intoxication. AB - Using a new ion-selective electrode, plasma concentration of ionized magnesium was measured in nine adult patients undergoing orthotopic liver transplantation. Baseline plasma ionized magnesium (IMg2+) concentration (0.49 +/- 0.07 mmol/L) was slightly below normal values (0.55-0.66 mmol/L, 95% CI): Six patients had ionized hypomagnesemia and two of these had total hypomagnesemia. Ionized IMg2+ concentration progressively decreased during the dissection (0.45 +/- 0.07 mmol/L, p < 0.05) and anhepatic stage (0.38 +/- 0.07 mmol/L, p < 0.05) and returned toward baseline values by 2 hours after graft reperfusion. Plasma ionized calcium levels and acid-base status were maintained within normal limits during surgery. Serum citrate concentration increased during the dissection (0.58 +/- 0.60 mmol/L) and anhepatic stages (1.18 +/- 0.78 mmol/L), the result of transfusion of citrate-rich blood products in the absence of adequate hepatic function, and gradually returned toward baseline values after graft reperfusion. IMg2+ concentration inversely correlated with the plasma citrate concentration (r2 = 0.54). The results of this study demonstrate that ionized hypomagnesemia invariably occurs during liver transplantation and suggest that this derangement may be a clinical concern, because magnesium is an important cofactor for the maintenance of cardiovascular homeostasis. The data further suggest the clinical importance of supplementation with magnesium based on the monitoring of plasma IMg2+ concentration. PMID- 9346675 TI - Preservation of cerebral oxidative metabolism in fulminant hepatic failure: an autoregulation study. AB - Under normal conditions cerebral blood flow (CBF) is regulated to secure oxidative brain metabolism, but in patients with fulminant hepatic failure (FHF), insufficient CBF has been suggested to precede cerebral edema and intracranial hypertension. In order to determine if insufficient CBF and hypoxia are present in patients with FHF we increased the mean arterial pressure and measured cerebral metabolism. In six patients with FHF CBF determined by 133Xenon injection technique, transcranial Doppler mean flow velocity in the middle cerebral artery (Vmean) and cerebral metabolism were determined, before and after an increase in mean arterial pressure by norepinephrine infusion. Mean arterial pressure was measured in a radial artery, and blood samples from the radial artery and internal jugular vein allowed calculation of the cerebral arteriovenous oxygen (AVDO2), -glucose (AVDgl), and -lactate (AVDlac) differences. Cerebral metabolic rates (CMRO2,-gl,-lac) were calculated as AVDO2, gl,-lac times CBF. Mean arterial pressure was raised from 70 (54-105) to 111 (93 128) mm Hg during intravenous infusion of norepinephrine. CBF increased from 34 (12-55) to 47 (27-81) mL . 100g-1. min-1 (p < 0.05) and Vmean from 53 (42-60) to 67 (61-79) cm.s-1 (p < 0.05), whereas CMRO2 (1.4 (0.9-2.4) mL . 100g-1 . min-1), CMRgl (11 (4.8-20) mumol 100g-1 . min-1), and CMRlac (3.2 (0-8.9) mumol . 100g-1 . min-1) remained unchanged. Our finding indicates that cerebral oxidative metabolism is preserved in patients with FHF. Cerebral autoregulation is absent, however, and neuroprotective critical care is suggested to be guided by internal jugular vein oxygen saturation to secure appropriate cerebral oxygenation. PMID- 9346676 TI - A percutaneous technique for venovenous bypass in orthotopic cadaver liver transplantation and comparison with the open technique. AB - Venovenous bypass minimizes the hemodynamic alterations during the anhepatic phase of liver transplantation. A new technique for the percutaneous placement of the bypass cannulae is described and compared to the cut-down ("open") technique. The records of 81 patients who underwent 94 liver transplants between August 1991 and April 1994 were reviewed for indications for transplant, United Network for Organ Sharing status, mean age, body surface area, bypass technique and time, flow rates, cardiac output, mean arterial pressure and central venous pressure during bypass, the development of deep venous thrombophlebitis, and lymphoceles. Femoral flow rates were higher in the open group (2054 +/- 74 mL/min), compared with the percutaneous group (1726 +/- 74 mL/min) (p = 0.003). Total flow rates in the open (2238 +/- 58 mL/min) and percutaneous (2197 +/- 67 mL/min) groups were not different. Maximum cardiac outputs (L/ min) were higher in the open (10.1 +/- 0.6) versus percutaneous group (7.0 +/- 0.5) (p < 0.0002). Similarly, minimum cardiac outputs (L/min) were higher in the open (8.9 +/- 0.7) versus percutaneous group (5.8 +/- 0.5) (p = 0.003). Other hemodynamic parameters (mean arterial pressure, central venous pressure) were not different between groups. Venous thrombosis occurred in 1/50 (2.0%) and 4/34 (11.8%) patients in the open and percutaneous groups, respectively (p = 0.153). Nineteen lymphoceles occurred in 102 (18.6%) at-risk sites in the open group, whereas no lymphoceles occurred in 66 at-risk sites in the percutaneous group (p < 0.001). Groin lymphoceles occurred in 7/50 (14%) and 0/34 at-risk sites in the open and percutaneous groups, respectively (p = 0.039). Axillary lymphoceles occurred in 12/52 (23.1%) and 0/32 at-risk sites in the open and percutaneous groups, respectively (p = 0.0031). Operative repair of a lymphocele was required in 11/16 (69%) patients. The percutaneous placement of catheters for venovenous bypass has the advantage of comparable flow rates with satisfactory hemodynamics without the lymphatic complications of the cut-down technique. PMID- 9346677 TI - Altered endothelin homeostasis in patients undergoing liver transplantation. AB - The liver is a major site of synthesis, clearance, and actions of the powerful vasoactive peptide endothelin-1 (ET-1). We investigated the role of the liver in ET-1 homeostasis by comparing circulating and hepatic ET-1 levels and hepatic ET receptors in patients undergoing orthotopic liver transplantation (OLTx) for end stage liver disease (ESLD) with those in patients undergoing liver resection for focal lesions with otherwise normal hepatic synthetic function. Central venous and radial arterial blood was drawn immediately after induction of anesthesia (point I), 10 minutes before beginning of resection or the anhepatic stage (point II), and 30 minutes after completion of resection or reperfusion of the grafted liver (point III). Portal and hepatic venous blood was drawn at points II and III. Plasma ET-1 levels were higher in ESLD patients than in resection patients. Plasma ET-1 levels rose both during resection and transplantation; the increase in ET-1 was more pronounced during transplantation. In ESLD patients, hepatic venous ET-1 was higher than portal venous ET-1, suggesting reduced clearance and/or enhanced synthesis of the peptide in the cirrhotic liver. Conversely, hepatic venous ET-1 was lower than portal venous ET-1 in resection patients at all time points and at point III in the ESLD patients. Hepatic concentration of ET-1 was greater and the capacity of the liver to catabolize ET-1 was reduced in ESLD patients as compared to the resection patients. Further, hepatic ET receptor density was higher in ESLD than in resection patients. These results suggest that the cirrhotic liver may contribute to elevated plasma ET-1 in ESLD. Considering its potent hemodynamic and metabolic effects in the liver, increased hepatic ET-1 and ET receptors and plasma ET-1 could play a role in the pathophysiology of liver disease and perioperative complications of OLTx. PMID- 9346679 TI - Development of monoclonal gammopathy precedes the development of Epstein-Barr virus-induced posttransplant lymphoproliferative disorder. AB - Epstein-Barr virus (EBV)-induced posttransplant lymphoproliferative disorder (PTLD) develops in 3% to 10% of solid organ transplant recipients with a resultant mortality of up to 70%. Unfortunately, there is no current marker which identifies patients who will develop this disease. We therefore conducted a risk factor analysis of variables that might predict the development of PTLD. Specifically, since EBV may cause both PTLD and the development of monoclonal proteins (M protein), we sought to determine if the development of an M protein preceded and therefore might serve as a predictive marker of subsequent PTLD. Before and after liver transplantation, 201 patients were evaluated for the presence of urine and serum M proteins. Patients were followed to monitor the development of PTLD for a mean of 1,733 days. PTLD developed in seven patients (3.5%), three (43%) of whom died from disseminated PTLD. PTLD was classified as polymorphous in six patients and monomorphous in one patient. Fifty-seven patients (28%) developed an M protein after transplantation: five of seven patients (71%) with PTLD and 52/194 (27%) of patients without PTLD. Multivariate risk factor analysis for the development of an M protein after transplantation identified cytomegalovirus (CMV) donor seropositivity (P = 0.0002) and postoperative symptomatic CMV infection (P = 0.019) as risk factors. Whereas EBV serostatus of either the donor or recipient was not found to be a risk factor for the occurrence of either an M protein or PTLD, the development of a serum immunoglobulin M (IgM) M protein (P = 0.04) and of any urine M protein (P = 0.01) was identified by univariate analysis as being associated with the development of PTLD. Further studies are needed to determine the predictive value of M proteins as a marker for PTLD. Until such time, the development of serum or urine M protein should heighten the suspicion of developing PTLD. PMID- 9346678 TI - Failure of ganciclovir prophylaxis to prevent allograft reinfection following orthotopic liver transplantation for chronic hepatitis B infection. AB - The effect of ganciclovir prophylaxis on reinfection of hepatic allografts by hepatitis B virus (HBV) was studied in 26 patients undergoing orthotopic liver transplantation (OLT) for decompensated cirrhosis due to HBV. Patients were randomized to receive either ganciclovir (6 mg/kg/day intravenously for a total of 100 days) or acyclovir (10 mg/kg every 8 hours intravenously until discharged and then 800 mg orally every 6 hours) for a total of 100 days after OLT as part of a study of prophylaxis against cytomegalovirus infection. All patients received hepatitis B immunoglobulin (HBIG), 10,000 units intravenously, during the anhepatic phase, daily for the first 7 days, after OLT, and then every 4 weeks for 6 months, Seven of 12 (58%) patients in the ganciclovir group developed recurrent HBV, compared with 6/14 (46%) of the acyclovir group (nonsignificant). No significant difference was observed in time to recurrent HBV in the ganciclovir group (mean 13.2 months) compared to the acyclovir group (mean 11 months). Our results suggest that ganciclovir administered prophylactically for 100 days after OLT does not prevent or delay graft reinfection by HBV. PMID- 9346680 TI - Destruction of Kupffer cells increases survival and reduces graft injury after transplantation of fatty livers from ethanol-treated rats. AB - This study investigated the role of Kupffer cells on survival and graft injury in transplanted fatty livers from rats treated acutely with ethanol. Donor rats were given ethanol (5 g/kg, by mouth) 20 hours before explantation, and liver grafts were preserved in University of Wisconsin cold storage solution for 24 to 42 hours prior to implantation. Blood samples were taken from the inferior vena cava for 3 hours after implantation. During this time, serum aspartate transaminase levels increased gradually from 122 U/L to 597 U/L in control rats, while ethanol treatment elevated values to 2,278 U/L. Gadolinium chloride (20 mg/kg, given intravenously to recipients 24 hours before explantation), a selective inactivator of Kupffer cells, minimized the increase in aspartate transaminase levels significantly. After implantation of grafts cold-stored for 42 hours, survival rates were 88% in control rats but only 33% in ethanol-treated rats. Gadolinium chloride improved survival nearly to control values. Ethanol nearly doubled white blood cell adhesion, an effect also largely blocked by gadolinium chloride. Further, alpha-(4-pyridyl 1-oxid)-N-tert-butylnitrone radical adducts detected in the bile were increased twofold by ethanol treatment. This effect was also reversed by gadolinium chloride. Taken together, these data indicate that survival is poorer and graft injury is greater in fatty livers from ethanol treated rats. Inactivation of Kupffer cells minimized graft damage, most likely by improving hepatic microcirculation and diminishing lipid peroxidation. PMID- 9346681 TI - Computed tomography versus magnetic resonance imaging--aided volumetry of the left lateral segment before living related liver donation: a case report. PMID- 9346682 TI - Detection of circulating donor deoxyribonucleic acid by microsatellite analysis in a liver transplant recipient. AB - The diagnosis of graft-versus-host disease following liver transplantation may be delayed because the clinical and pathological features are nonspecific. We report the use of microsatellites to support a diagnosis of GVHD in a patient who developed fever and a skin rash 28 days after liver transplantation. The pattern of microsatellite alleles amplified from the peripheral blood on day 51 posttransplant indicated that recipient and donor DNA were present in approximately equal proportions. Microsatellite typing is a simple and rapid method to identify high levels of circulating donor DNA to support a diagnosis of GVHD following liver transplantation. PMID- 9346683 TI - The A to Z of new hepatotropic agents: human hepatitis viruses and monkey business. PMID- 9346684 TI - UNOS Center specific data. PMID- 9346685 TI - Long-term postoperative care of the liver transplant patient. PMID- 9346686 TI - Long-term immunosuppression without corticosteroids after orthotopic liver transplantation: a positive therapeutic aim. AB - Long-term treatment with corticosteroids after orthotopic liver transplantation (OLT) may cause adverse effects, particularly hypertension, diabetes, and bone disease. The results of steroid withdrawal from long-term immunosuppression in 114 patients after OLT was reviewed. Initial treatment was with corticosteroids, azathioprine, and cyclosporine A in 76.3% and with antithymocyte globulin in 17.5%. Corticosteroids were stopped in 96 patients (84.2%) during mean follow-up of 6.7 +/- 3.9 months, and acute rejection subsequently developed in 8. By comparison 7 of 18 patients, in whom corticosteroids were continued, developed acute rejection. Six of these had received blood group (ABO)-compatible nonidentical grafts. Rates for retransplantation in the steroid withdrawal and nonwithdrawal groups were 4.2% and 22.2%, respectively, and mortality in the two groups was 14.6% and 44.4%, respectively. Azathioprine was not given or withdrawn in 28 patients in the group from which corticosteroids were also withdrawn, with no adverse effect. Diabetes mellitus improved following corticosteroid withdrawal, but there was no improvement in hypertension. We conclude that corticosteroids can be safely withdrawn in the majority of patients after OLT. PMID- 9346688 TI - Morbidity and mortality in patients with coronary artery disease undergoing orthotopic liver transplantation. AB - Thirty-two patients with coronary artery disease who underwent liver transplantation between 1990 and 1994 were identified. Coronary artery disease was managed medically (n = 9), by angioplasty (n = 1), or surgically (n = 22) prior to liver transplantation. Two patients underwent simultaneous coronary artery bypass grafting and liver transplantation. Complete preoperative cardiac evaluation was performed in all patients. Perioperative and postoperative morbidity and mortality were retrospectively determined. Overall mortality was 50%, whereas morbidity was 81%. Follow-up was between 1 and 3 years after liver transplantation. Subgroup analysis revealed that medically managed patients had a 56% mortality and a 100% morbidity. The patient who underwent angioplasty survived without morbidity. One patient who underwent simultaneous coronary artery bypass grafting and liver transplantation died intraoperatively. The second patient survived but required pacemaker insertion and inotropic agents postoperatively. The 20 patients with prior coronary artery bypass grafting had a 50% mortality and 80% morbidity. Further, analysis by United Network for Organ Sharing functional status revealed a higher than expected mortality in all groups. The morbidity and mortality associated with liver transplantation is significantly increased in patients with coronary artery disease and is equally high in medically and surgically treated patients. By comparison, patients without coronary artery disease have a 3-year survival of 55.4% (status I) to 79.7% (status III and IV). The increased intraoperative and postoperative risk in patients with coronary artery disease undergoing liver transplantation should be considered when determining the candidacy of these patients as well as when providing informed consent. PMID- 9346687 TI - Liver transplantation in HBsAg-positive HBV-DNA--negative cirrhotics: immunoprophylaxis and long-term outcome. AB - The presence of a positive hepatitis B surface antigen (HBsAg) has been considered a highly questionable indication for orthotopic liver transplantation. We report our experience in the treatment of HBsAg-positive HBV-DNA-negative cirrhotics with liver transplantation, whether or not followed by passive prophylaxis with specific immunoglobulins. Of the 123 cirrhotics who received transplants at our institution since May 1986, 39 (31.7%) were HBsAg positive; of these, 1 was HBV-DNA positive, and 4 were hepatitis Be antigen (HBeAg) positive. Since April 1991, 25 HBsAg-positive HBV-DNA-negative cirrhotics have undergone an original protocol with the periodical intramuscular administration of 5,000 IU of specific immunoglobulins starting in the anhepatic phase and lasting for at least 1 year. There were no differences among cirrhotics in terms of operative mortality and long-term survival with respect to the presence of the HBsAg. Of the 35 HBsAg-positive HBV-DNA-negative patients having a follow-up of 1 month or longer, 12 (34.3%) developed HBsAg recurrence; of them, 4 (33.3%) had received a complete prophylaxis, whereas 8 (66.7%) had not. The recurrence rate was 80% (8 out of 10) in the group of patients who had not received the prophylaxis and 16% (4 out of 25) in the group who had received the prophylaxis (P = .0003). The actuarial recurrence rate in the treated patients was 20.2% and 20.2% after 1 and 3 years, respectively, whereas in the untreated group it was 60.0% and 70.0% (P < .01). The hazard of recurrence of treated patients was reduced to 24.9% compared with untreated patients. Liver transplantation can be performed in HBsAg-positive HBV DNA negative patients without an increase in the operative risk or a worsening of long-term results. Immunoglobulin prophylaxis seems to be effective in preventing hepatitis recurrence after transplantation. PMID- 9346689 TI - Influenza vaccination following liver transplantation in children. AB - Our objective was to determine the immunologic response to two influenza vaccine doses in 39 children who had undergone liver transplantation. Patients received two doses of trivalent inactivated influenza vaccine 4 weeks apart. Sera were collected 4 weeks after each dose and analyzed by a hemagglutination inhibition assay (HAI) for evidence of antibody response to the antigens A/Taiwan/1/86 (H1N1), A/Beijing/32/92 (H3N2), and B/Panama/45/95. Patients with HAI titers of 1:40 or greater were considered to have protective titers. Twenty-six (67%) patients showed a 1:40 or greater titer response to A/Beijing/32/92 1 month after the first vaccination. Only two additional patients were found to have similar titers after the second dose. A higher proportion of patients with protective titers were on smaller amounts of prednisone for body weight or alternate day low dose (< 10 mg/day) prednisone compared to patients on daily low dose or daily high dose prednisone. Patients with protective titers were significantly older (9.0 +/- 2.8 years) than those without protective titers (4.2 +/- 3.4 years, p = .002) following the first inoculation of the A/Beijing/32/92 vaccine component. Similar results were found for the second vaccination and with the H1N1 antigen. Cyclosporine level, gender, and body mass index were not associated with any outcome measures. We conclude that most liver transplant recipients had a protective antibody titer after a single influenza inoculation, but little further advantage was gained after an additional dose. Vaccination of household contacts of younger patients and those patients on daily prednisone or patient chemoprophylaxis may offer greater benefit in prevention of influenza in liver transplant recipients than multiple vaccine doses with current vaccine preparations. PMID- 9346690 TI - Accuracy and significance of pretransplant liver volume measured by magnetic resonance imaging. AB - Measurement of liver volume in patients with advanced liver disease is used to gauge the appropriate size of donor organs and may have prognostic value. We sought to determine the accuracy of magnetic resonance imaging (MRI) in measuring liver volume in 19 adult patients under consideration for liver transplantation. We also correlated the liver volume determination to the clinical severity of disease. Liver volume was measured at MRI by averaging the calculated volumes from coronal and transverse breath-hold T1-weighted images. These results were compared to the explanted liver volume measured by fluid displacement and the explant mass. The correlation coefficient for MRI liver volume and the explant displacement volume was 0.90. The mean liver volume for Child-Pugh class AB by MRI was 1986 +/- 568 mL (1002-2470 mL) compared to 1433 +/- 379 mL (540-1889 mL) in Child-Pugh class C patients (p = .02). We conclude that MRI offers an anatomically accurate means of determining adult liver volume in vivo. Lower mean liver volumes were observed in Child-Pugh class C patients. In addition to its ability to provide tumor screening and vascular assessment, MRI is able to provide accurate determinations of liver volume in patients undergoing liver transplant evaluations. PMID- 9346691 TI - Glucose and potassium metabolic responses to insulin during liver transplantation. AB - Insulin regulates glucose and potassium metabolism by acting differently upon peripheral tissues (e.g., skeletal muscle) and the splanchnic bed, including the liver. Liver disease is accompanied by "insulin resistance" of glucose metabolism, whereby glucose intolerance occurs despite relatively increased plasma insulin concentration. However, it is unknown whether insulin resistance extends to potassium metabolism. Further, it is uncertain whether the hyperglycemia and alterations of plasma potassium concentration observed during liver transplantation result from changes in circulating insulin concentration, altered sensitivity to insulin, or both, as the diseased liver is removed and replaced with a graft organ. The present study evaluated the role of the liver in maximal insulin responsiveness of whole-body glucose and potassium metabolism, using a hyperinsulinemic clamp technique, to identify the mechanism(s) underlying post-reperfusion hyperglycemia and intraoperative hyperkalemia. Two protocols were employed: in protocol 1 (n = 10), no exogenous insulin was administered. In protocol 2 (n = 10), an intravenous insulin bolus (666 mU . kg-1) was administered after anesthesia induction, followed by an infusion at 500 mU.m 2.min-1, which continued until 3 hours after portal vein unclamping. Plasma concentrations of glucose and potassium were regulated by glucose and potassium chloride infusion (euglycemic eukalemic clamp). Insulin-stimulated exogenous glucose and potassium uptakes were determined in protocol 2 before skin incision and during the dissection, anhepatic, and neohepatic stages. In both protocols, serial measurements of hemodynamic arterial blood gases, glucose, free fatty acids, potassium, insulin, and glucagon concentrations were made. Without insulin (protocol 1), progressive hyperglycemia peaked after portal vein unclamping (post reperfusion hyperglycemia), with no concomitant decrease in plasma insulin concentration. Intraoperative plasma potassium concentration did not change. Insulin infusion (protocol 2) produced a stable hyperinsulinemia (approximately 2000 microU/mL). Hyperinsulinemia did not eliminate post-reperfusion hyperglycemia. Insulin-stimulated glucose uptake, in mg . kg-1 . min-1, was 8.10 +/- 0.78 (mean +/- SE) before skin incision, 7.62 +/- 0.82 during the hepatic dissection, 4.40 +/- 0.75 during the anhepatic stage, and 4.06 +/- 0.74 at 3 hours after portal vein unclamping. Insulin-stimulated potassium uptake, in mEq . kg-1 . hr-1, was 0.24 +/- 0.02 before skin incision, 0.21 +/- 0.04 during hepatic dissection, 0.07 +/- 0.02 during the anhepatic stage, and 0.21 +/- 0.04 and 0.19 +/- 0.05 at 30 minutes and 3 hours, respectively, after portal vein unclamping. We conclude that post-reperfusion hyperglycemia is not due to inadequate insulin stimulation. Liver disease-induced insulin resistance of glucose metabolism is exacerbated by hepatectomy and is not reversed during the intraoperative neohepatic stage. Liver disease does not impair maximal insulin-stimulated potassium uptake. The liver, even with end-stage disease, accounts for approximately 70% of insulin-stimulated potassium uptake. PMID- 9346692 TI - Nitroglycerin versus epoprostenol: effects on hemodynamics, oxygen delivery, and hepatic venous oxygenation after liver transplantation. AB - Our objective was to determine the effects of vasodilatory treatment with epoprostenol (PGI2) and nitroglycerin (NTG) on systemic oxygen delivery index (DO2) and hepatic venous oxygen saturation (SvhO2) after liver transplantation. This prospective study used repeated-measures design. Fifteen adult patients undergoing orthotopic liver transplantation (OLT) were enrolled. Postoperatively, a fiberoptic pulmonary artery catheter was inserted into the right hepatic vein and a timed infusion of PGI2 and NTG was sequentially performed in random order at the following rates: PGI2 at 5 ng/kg/minute and NTG at 0.1 microgram/kg/minute. Each step in each sequence lasted 45 minutes, followed by a control interval of 45 minutes. Measurements were taken at the end of each period when hemodynamic function was stable. Systemic hemodynamics, DO2, oxygen uptake index (VO2), mixed venous oxygen saturation (SvO2), and SvhO2 were assessed. We found that PGI2 induced an increase of cardiac index (+18%, p < .05); DO2 (+16%, p < .05); and SvhO2 (+11%, p < .05). Mean arterial pressure was decreased during PGI2 infusion (-9%, p < .05), as well as during infusion of NTG (-10%, p < .05). NTG significantly decreased DO2 (-6%, p < .05) and SvhO2 (-4%, p < .05). Neither drug affected VO2. We conclude that PGI2 induced vasodilation and increased systemic oxygen delivery in parallel with SvhO2, suggesting a corresponding increase of hepatic oxygen supply. NTG induced systemic vasodilation and significantly impaired hepatic venous oxygen saturation and DO2. Thus, if vasodilatory therapy is indicated in the patient after liver transplantation, PGI2 appears to be better than NTG in improving DO2 without impairing splanchnic oxygenation. PMID- 9346693 TI - Effect of total hepatic vascular exclusion during liver resection on hepatic ultrastructure. AB - The aim of this investigation was to observe ultrastructural changes in the liver in response to warm ischemia during liver surgery. In 11 noncirrhotic patients, hepatic resection was performed under total vascular exclusion (TVE). The mean duration of warm ischemia was 28 minutes (range 16-48 minutes). Three specimens were taken from each patient: before clamping, at the end of TVE, and after reperfusion. Biopsy specimens were studied by light microscopy and by transmission electron microscopy (EM). At the end of the ischemic phase sinusoids were collapsed with resultant loss of normal hepatic architecture. Morphological changes to hepatocytes included focal chromatin condensation at the nuclear margins, distended nuclear envelope, and swelling of both mitochondria and endoplasmic reticulum. After reperfusion these changes reversed. The phenomenon of sinusoidal and hepatocellular recovery after TVE was seen in all the cases of this study, irrespective of age, sex, disease, type and severity of surgical intervention, and duration of TVE. It can be concluded that TVE over a period of 48 minutes has no irreversible deleterious effects on the ultrastructure of the noncirrhotic liver. PMID- 9346694 TI - Presentation of choledochal cysts without intrabiliary communication on endoscopic retrograde cholangiopancreatography. AB - Choledochal cysts have generally been described to communicate with the biliary system. We recently saw three adult patients in whom we could not demonstrate biliary communication between the cyst and the biliary tree, despite a carefully performed endoscopic cholangiopancreatography (ERCP). These cases may represent a variant of type II choledochal cysts. During the 5-year period of review of ERCP records, 30 choledochal cysts were diagnosed, of which 3 (10%) were the noncommunicating cases described in this report. PMID- 9346695 TI - Retransplantation for precore mutant-related chronic hepatitis B infection: prolonged survival in a patient receiving sequential ganciclovir/famciclovir therapy. AB - Retransplantation for hepatitis B-related liver allograft failure is rarely successful. Recurrence of infection is almost universal, and the second allograft is invariably lost more rapidly than the first. In a recent multicenter study, only 1 of 20 hepatitis B virus (HBV)-positive patients who underwent liver retransplantation survived beyond 6 months. This report describes the long-term effect of antiviral therapy in a 56-year-old man who was retransplanted for HBV related allograft loss 14 months after his initial liver transplant. He was treated after the second transplant with intravenous daily ganciclovir. After 10 months of this therapy HBV recurrence was detected. After a change to oral famciclovir therapy, there was a decrease in serum HBV DNA and amino-transferase levels and an improvement in the patient's clinical condition. Famiciclovir therapy has now been continued for 26 months, and the patient remains well 3 years after his second transplant, despite persistent HBV infection and progression to cirrhosis. These observations indicate that the use of long-term antiviral therapy offers promise for improving outcomes in patients who undergo retransplantation after HBV-related liver allograft failure. PMID- 9346696 TI - Mercury poisoning associated with high-dose hepatitis-B immune globulin administration after liver transplantation for chronic hepatitis B. PMID- 9346697 TI - Overall evaluation: screening and assessment of risk factors. PMID- 9346698 TI - The older liver transplant candidate: what are the limits? PMID- 9346699 TI - Assessment of the alcoholic patient for liver transplantation: comorbidity, outcome, and recidivism. PMID- 9346701 TI - Liver transplant evaluation: linking risk factors and outcome in the patient with variceal bleeding. PMID- 9346700 TI - The candidate in the intensive care unit: assessing risk. PMID- 9346702 TI - The patient with renal insufficiency. PMID- 9346703 TI - Risk factors and outcomes after pediatric liver transplantation. PMID- 9346704 TI - The living-related liver transplant evaluation: linking risk factors and outcome. PMID- 9346705 TI - The patient with chronic hepatitis B. PMID- 9346706 TI - The patient with chronic hepatitis C. PMID- 9346707 TI - Nutritional assessment and therapy in patients requiring liver transplantation. AB - Pretransplant nutritional assessment in the patient with ESLD is problematic. The best system for nutritional assessment uses a "global" evaluation of the patient's nutritional reserves. With such a technique, the vast majority of transplant candidates have been shown to have evidence of malnutrition. Several investigators have demonstrated the risk of significant malnutrition on posttransplant outcome. An aggressive approach to nutritional repletion is necessary to improve the ESLD patient's metabolic reserves, maintain remaining hepatic function, and better the outcome after liver transplantation. PMID- 9346708 TI - Prevention of infection in the liver transplant recipient. PMID- 9346709 TI - Liver transplantation for primary sclerosing cholangitis: impact of risk factors on outcome. AB - The results of liver transplantation in patients with PSC are excellent and the quality of life is markedly improved. Indeed, liver transplantation is the therapy of choice for patients with end-stage PSC. However, in an age of cost containment, it appears that there are several advantages to offering transplant to patients with PSC a little bit earlier rather than later in the course of their disease. It appears that we can further improve survival, decrease morbidity, decrease blood usage, and avoid the risk of developing a cholangiocarcinoma, which occurs sporadically but not infrequently in the PSC patient. In addition, avoidance of right upper quadrant surgery, such as biliary or shunt surgery, appears to offer several advantages by decreasing resource utilization and possibly decreasing mortality. Although the UNOS selection guidelines recommend transplantation of the sickest patient, there appears to be accumulating evidence that transplantation in patients earlier in the course of their end-stage liver disease may improve survival, decrease morbidity, and also importantly, decrease the cost associated with this expensive procedure. Ideally, we would recommend consideration for liver transplantation all PSC patients who have (1) a Mayo risk score of > 4.8 in whom malignancy is ruled out, (2) cirrhosis and complications of portal hypertension such as variceal bleeding, refractory ascites, or portosystemic encephalopathy, or (3) disabling symptoms such as fatigue, pruritus, or recurrent bacterial cholangitis. We believe that biliary surgery to treat dominant strictures should be avoided and that such strictures should be approached either endoscopically or radiographically, which should include brushings, biopsies, and histology to reasonably exclude the diagnosis of cholangiocarcinoma. Finally, we continue to search for risk factors and for early markers of cholangiocarcinoma so these patients can be identified early and this devastating complication can be avoided by early transplantation. PMID- 9346710 TI - Evaluation of the candidate with a previous malignancy. PMID- 9346711 TI - The surgical treatment of primary hepatobiliary malignancy. PMID- 9346722 TI - Changes in quality of life after liver transplantation among adults. National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Liver Transplantation Database (LTD). AB - Quality of life is an important factor to consider when assessing the value of liver transplantation. Using a large, prospective database of liver transplantation recipients from three clinical centers in the United States, we examined the quality of life of 346 adults before and 1 year after surgery. Five quality of life domains were evaluated (measures of disease, psychological distress and well-being, personal function, social/role function, and general health perception) with standardized questionnaires completed according to established protocol. The largest numbers of patients were distressed by fatigue and muscle weakness, both before transplantation and 1 year after surgery. Compared to baseline, recipients at follow-up noted fewer disease-related symptoms (P < .001) and lower levels of distress overall (P < .001). However, levels of distress due to excess appetite (P < .001), headaches (P = .02), and poor/blurred vision (P = .05) were more likely to increase than decrease. Although 57% to 64% of the recipients were distressed by each of the psychological conditions examined at follow-up, distress was more likely to decrease than increase (P < .001), and well-being was comparable to the general population. All measures of personal functioning improved significantly (P < .05). Fifty-eight percent of the patients prevented by their disease from going to work or school before transplantation were no longer so limited at follow-up. With the exception of marriage (P = .23), all facets of social/role functioning improved more often than worsened (P < .01). Perception of health improved remarkably, with 13.4 times as many recipients reporting improved health as reporting worse health (P < .001). We conclude that liver transplantation markedly improves the quality of life of patients with end-stage liver disease. PMID- 9346723 TI - The clinical course of transplantation-associated de novo hepatitis B infection in the liver transplant recipient. AB - Transmission of hepatitis B infection from hepatitis B surface antigen (HBsAg) negative antibody to hepatitis B core (anti-HBc)-positive liver donors has been previously described. The long-term outcome of these transplant-associated de novo hepatitis B patients has not been well described and may affect future use of donor organs that are anti-HBc-positive. We describe the experience in our first 332 transplants, performed before exclusion of anti-HBc-positive liver donors. Nine of these 332 (3%) donors were anti-HBc positive. Three of these 9 (33%) recipients developed transplant-associated de novo hepatitis B infections compared with only 2 of 323 (0.5%) recipients who received anti-HBc-negative donor livers (P = .00014). Of our 9 recipients of anti-HBc-positive livers, 6 (67%) are alive, and no deaths or allograft failures have been related to complications of hepatitis B. Only 1 of 5 patients (20%) with de novo hepatitis B has developed significant graft dysfunction with an average follow-up of more than 7 years (range 63-124 months). The 1 recipient with allograft dysfunction related to de novo hepatitis B has significantly elevated viremia levels (average HBV DNA 460 pg/mL) compared with the other de novo hepatitis B recipients (average HBV DNA 23-58 pg/mL). In summary, anti-HBc-positive donors are more likely to transmit hepatitis B infections to recipients, but these de novo infections usually have a mild clinical course and do not seem to adversely affect long-term patient survival. Hepatitis B-related allograft dysfunction, when it occurs, is associated with higher levels of viral replication. With our current donor shortage, perhaps anti-HBc-positive donors could be used in very selected recipient populations. PMID- 9346724 TI - Peripheral eosinophil count both before and after liver transplantation predicts acute cellular rejection. AB - Acute cellular rejection is common after orthotopic liver transplantation and an important cause of graft dysfunction. Eosinophils, potent mediators of tissue damage, have been implicated in the pathogenesis of acute rejection. We studied 55 patients, all of whom had a protocol biopsy 7 days after transplantation and whose peripheral eosinophil count was monitored daily for 11 days after transplantation. Patients were divided clinicopathologically into two groups: group A, without rejection, group B, with rejection. Group B (36% of patients) developed rejection within the 11-day study period. The pretransplant eosinophil count was significantly higher in group B, compared with group A (0.31 +/- 0.08 v 0.10 +/- 0.01 (x10(9)/L), p < .001). After transplantation, the eosinophil count fell to low levels in both groups. By day 3 there was a statistically significant rise in the eosinophil count in group B compared with group A, with a maximum at day 7 [0.51 +/- 0.06 v 0.26 +/- 0.03 (x10(9)/L) p < .001]. After treatment with steroids, the eosinophil count dropped to values similar to those in group A and remained low thereafter in 16 of 20 patients. Four patients had a second episode of rejection; in each of these, eosinophils were raised again and decreased with resolution of the rejection. An eosinophil count threshold of 0.13 (x10(9)/L) before transplantation and 0.33 (x10(9)/L) on day 7 after transplantation predicted the development of rejection (sensitivity 72/70%, specificity 66/63%, negative predictive value 82/79%). We conclude that a raised eosinophil count is associated with acute rejection. The raised eosinophil count before transplantation in group B suggests that these patients are predisposed to acute rejection, and earlier intervention may be indicated. PMID- 9346725 TI - Interleukin-2 and interleukin-12 mediate distinct effector mechanisms of liver allograft rejection. AB - Interleukin-2 (IL-2), interleukin-12 (IL-12) or interleukin-4 (IL-4) were administered postoperatively to otherwise spontaneously accepting mouse liver allograft recipients (C57BL/10-->C3H) to test whether TH1 cytokines are critical mediators of rejection in this model. The induction of rejection at days 5 to 7 by exogenously administered IL-2 and IL-12, but not IL-4, suggests that mouse liver allograft rejection can be induced by TH1 cytokines; however, there appeared to be differences in the mechanism by which these cytokines induce liver rejection. IL-2 administration was accompanied by an increased intragraft infiltration of CD4+ and CD8+ cells and an up-regulation of natural killer (NK), lymphokine-activated killer (LAK), allospecific cytotoxic killer (CTL) activity and perforin mRNA when compared with media-treated controls. In contrast, exogenous IL-12 treatment was associated with a suppression of CTL, NK, and LAK activity compared with controls but an enhanced infiltration of F4/80+ macrophages as determined by immunohistochemistry. Determination of cytokine mRNA profiles by semi-quantitative reverse transcription polymerase chain reaction showed the up-regulation of interferon (IFN)-gamma, IL-4, IL-6, and IL-10 mRNA with IL-2 treatment when compared with media-treated controls. Interestingly, IL 2 mRNA was down-regulated in these animals, suggesting a negative feedback mechanism in IL-2 regulation. IL-12 treatment resulted in the up-regulation of IFN-gamma, IL-6, and IL-10 mRNA, but not IL-2 or IL-4 mRNA. Higher complement directed cytotoxic antibody titers were seen in IL-12-treated recipients compared with controls, whereas IL-2 treatment showed no apparent differences in antibody titers compared with media treatment. These in vivo observations were mimicked in a mixed leukocyte reaction by supplementing the reaction with IL-2, IL-12, or media. These results suggest that rejection of mouse liver allografts may involve more than one distinct cellular immunological effector mechanism. One is mediated by IL-2 and appears to favor alloreactive CTL, whereas the other pathway is mediated by IL-12/IFN-gamma and involves macrophages and cytotoxic antibodies largely resembling a delayed-type hypersensitivity reaction. PMID- 9346726 TI - Retransplantation for recurrent hepatitis C. AB - Recurrence of hepatitis C virus (HCV) after orthotopic liver transplant (OLT) may be mild or may lead to progressive liver disease requiring retransplantation (re OLT). Results of re-OLT for hepatitis C are not well known. We analyzed outcomes in 14 patients retransplanted for recurrent hepatitis C. All had evidence of recurrent hepatitis on multiple biopsies. Polymerase chain reaction (PCR) was performed in blood or tissue samples from 12 patients when recurrence was suspected; all 12 were positive for HCV-RNA. Explants showed chronic hepatitis with bridging necrosis in 3 patients, hepatitis with transition to cirrhosis in 2, hepatitis and cirrhosis in 3, and cirrhosis alone in 2. In 2 patients, in whom immunosuppression had been withheld for 4 to 6 weeks, there was also evidence of chronic rejection. Four died of sepsis perioperatively (median, 32.5 days; range, 9-59); pre-OLT, 3 of 4 had renal failure, and 1 had fever with no obvious source of infection. Ten patients did well early after OLT and were discharged. One patient was readmitted 6 weeks after discharge and died of cytomegalovirus (CMV) infection 127 days after re-OLT. One patient with concomitant vanishing bile duct syndrome, probably due to chronic rejection, developed recurrent hepatitis and died of progressive liver failure 161 days after re-OLT. Eight patients are well at a median of 926 days (range, 315-1930) after re-OLT. Three have evidence of mild recurrent hepatitis on liver biopsy, one is overweight with severe steatosis on biopsy, and four have no evidence of recurrent hepatitis. Retransplantation for hepatitis C should be considered a viable option for patients who develop end stage hepatic dysfunction secondary to recurrent disease and should be performed before development of infectious complications and renal insufficiency. PMID- 9346727 TI - Primary graft dysfunction after liver transplantation: from pathogenesis to prevention. PMID- 9346728 TI - Methemoglobinemia associated with dapsone treatment in solid organ transplant recipients: a two-case report and review. AB - Dapsone, a sulfone antibiotic, has been increasingly used in solid-organ transplant recipients for the primary prevention of Pneumocystis carinii pneumonia, especially in patients with documented sulfa allergy. A known side effect of dapsone therapy, however, is methemoglobinemia, a condition leading to impaired tissue oxygen delivery. This report documents two cases of dapsone induced methemoglobinemia in patients after solid organ transplantation with emphasis on the importance of clinical recognition and benefits of treatment. Further, the pathophysiology and causes of this condition are extensively reviewed. PMID- 9346729 TI - Lower body impedance for the evaluation of venovenous bypass flow. AB - Inferior vena cava (IVC) clamping during liver transplantation causes venous congestion in the splanchnic and IVC beds. A venovenous bypass relieves congestion and improves cardiac output (CO), but the bypass flow required for adequate drainage of the vascular beds is controversial. In this study we evaluated the bypass flow necessary to compensate for the IVC clamping. Lower body impedance (BI) is inversely related to tissue fluid content and was used to reflect congestion. A venovenous bypass was successfully applied to 59 of 62 patients. BI was measured across the left buttock and related to bypass flow, CO, bypass flow ratio (bypass flow/CO before IVC clamping; n = 62), and right femoral venous pressure (n = 8). The bypass flow was 1.7 (0.0-3.0) L.min-1 (median and range). BI decreased (delta BI; -2.2 [-10.3-1.1)] omega) as the femoral venous pressure increased (29 [21-49] mm Hg; r = -0.81; P < .05), and the femoral venous pressure correlated inversely to bypass flow (r = -0.35; P < .01). The change in CO at IVC clamping (delta CO; -2.3 [-6.3-1.6] L.min-1) related to bypass flow ratio (0.25 [0-0.51]; r = 0.57, P < .01), whereas delta BI related only minimally to bypass flow or bypass flow ratio (r = 0.37; P < .05). In conclusion the median bypass flow of 1.7 L.min-1 was too small to prevent fluid accumulation in the lower caval region, and extrapolation of data suggests that bypass flow should have approached 3.5 L.min-1 or 50% of CO in order to prevent fluid accumulation in the lower caval region. However the minimal correlation between lower BI and bypass flow indicates that bypass flow per se is not the only determinant of lower body fluid accumulation. PMID- 9346730 TI - Hepatic adenoma and focal nodular hyperplasia: diagnosis and criteria for treatment. AB - Focal nodular hyperplasia (FNH) and adenoma are rare benign hepatic tumors, and the standards for diagnosis and treatment still remain controversial. Usually adenoma is an indication for resection, due to its tendency to bleed and to degenerate; FNH, on the contrary, may be treated conservatively. Preoperation differential diagnosis is, however, difficult, often impossible. MATERIALS AND METHODS: Thirty-eight patients with presumed hepatic adenoma and/or FNH were studied at our department from 1984 to 1996. Preoperative assessment included clinical evaluation and symptoms, laboratory tests, liver biopsy, ultrasound scan, computed tomography scan, magnetic resonance imaging, scintigraphy, and angiography. Thirteen patients had a presumed diagnosis of FNH, 16 of adenoma, and 9 of undetermined benign lesions; 27 had hepatic resections (3 with laparoscopic technique), and 11 were not operated on and are actually under a strict follow-up observation. RESULTS: The final diagnosis was 19 FNH and 19 adenomas (2 of which contained areas of hepatocarcinoma). Presumed diagnosis was confirmed in 71% of cases. Use of oral contraceptives, abdominal symptoms, and pathologic liver test results were frequent in patients with adenomas. There were no deaths after surgery. All resected patients were tumor free during the follow up, and in 10 of the 11 nonoperated cases, the size of the nodules remained unchanged. We conclude that precise diagnosis of these benign liver tumors remains difficult and sometimes impossible, despite new imaging techniques. Hepatic resections can be performed under very safe conditions; laparoscopic surgery may play a role in selected cases. Adenomas and uncertain cases are clear indications for surgery. Only when a diagnosis of FNH can be firmly confirmed in asymptomatic patients is strict observation without surgery recommended. PMID- 9346731 TI - Quantitative liver function tests define the functional severity of liver disease in early-stage cirrhosis. AB - Some patients with early-stage cirrhosis preserve hepatic function, whereas others have little hepatic reserve and rapidly deteriorate. The aim of this study was to use quantitative tests of liver function (QLFTs) to define the degree of functional hepatic impairment in patients with early-stage cirrhosis (Child-Pugh score 5-7) and to determine whether the tests predicted subsequent hepatic decompensation. We recruited 10 cirrhotic (Cr) patients and 10 healthy controls (NI), who were well matched for race, age, weight, and gender. Clearances of caffeine (CF) and antipyrine (AP) after oral administration were measured from timed samples of saliva. The clearance of cholate (CA) was measured from serum samples obtained after simultaneous oral ([2,2,4,4-2H]CA) and intravenous ([24 13C]CA) administration. CA shunt was calculated as (Cl i.v./Clo x 100%). CF elimination rate (Cr v NI, mean +/- SD: 0.03 +/- 0.02 v 0.075 +/- 0.018 h-1, P < .0005) and AP clearance (24 +/- 16 v 40 +/- 7 mL/minute, P < .02) were reduced in Cr patients. CA shunt was increased in Cr patients (43 +/- 18 v 18 +/- 7%, P < .002). Five Cr patients decompensated during follow-up and had the worst CA shunts (76%, 66%, 51%, 48%, and 45%). Three subsequently received successful orthotopic liver transplantation, 1 died of hepatoma, and 1 is on the waiting list for transplantation. In conclusion, QLFTs define the degree of functional impairment in early cirrhosis and may identify Cr patients at greatest risk of decompensation who may require transplantation for survival. PMID- 9346732 TI - Recurrence of nonalcoholic steatohepatitis in a liver transplant recipient. AB - A 42-year-old white man with morbid obesity and hypertriglyceridemia was noted to have nonalcoholic steatohepatitis (NASH) at the time of a laparoscopic cholecystectomy for presumed gallstone pancreatitis. His postoperative course was complicated by a 50-kg weight loss and continued right upper quadrant pain. Repeat liver biopsy revealed NASH with accompanying micronodular cirrhosis. Due to progressive fatigue, he underwent an orthotopic liver transplantation complicated by a 36-kg weight gain. Sixteen months posttransplantation, a liver biopsy revealed the recurrence of NASH. Screening for defects in fatty acid oxidation proved negative. PMID- 9346733 TI - Recurrent nonalcoholic steatohepatitis and cirrhosis after liver transplantation. AB - Nonalcoholic steatohepatitis (NASH) may progress to cirrhosis and lead to liver failure. Histologically, NASH is often indistinguishable from liver disease caused by alcohol use; the cause of NASH remains unknown. A subgroup of patients with NASH eventually develops fibrosis and/or cirrhosis, and in many cases, transplantation is performed for end-stage liver disease attributed to steatohepatitis in patients who do not consume alcohol. The patient described received a transplant for end-stage liver disease secondary to NASH with cirrhosis. Postoperatively she did well, with a bout of mild rejection treated successfully at week 9 with prompt normalization of liver tests. Weight and glycemic control were optimized, and steroid therapy was minimized as safely as possible. Repeat liver biopsy at week 66, however, for persistent mild elevation of alkaline phosphatase and gamma-glutamyl-transferase surprisingly revealed the "recurrence" of NASH. Subsequent biopsy revealed NASH with cirrhosis by week 76 after transplantation. Subsequent biopsy at week 87 has confirmed cirrhosis. The patient does not consume alcohol. It is believed to be the first reporting of such a case. PMID- 9346734 TI - The role of the host immune state in recurrent hepatitis C. PMID- 9346735 TI - Hard times and imperfect organs. PMID- 9346736 TI - Recurrence of nonalcoholic steatohepatitis after liver transplantation. PMID- 9346737 TI - Papillary neoplasia of the biliary tract. PMID- 9346738 TI - Winging it. PMID- 9346739 TI - Milestones in liver transplantation for alcoholic liver disease. PMID- 9346740 TI - The etiology, consequences, and treatment of alcoholism. PMID- 9346741 TI - Alcoholic liver disease: natural history. AB - Alcohol has been implicated in the genesis of liver disease for centuries. Modern epidemiological data from many societies corroborate the correlation between per capita consumption of alcohol and deaths from cirrhosis. Although significant progress has been made in our understanding of the pathogenesis of alcoholic liver disease, ALD, effective therapies for most individuals with ALD have not been found. High per capita consumption of alcohol, coupled with the dearth of effective treatments and the failure of most affected individuals to abstain from alcohol, explains why ALD is one of the most prevalent forms of disabling, chronic liver disease. PMID- 9346742 TI - Liver transplantation for alcoholic liver disease in the United States: 1988 to 1995. PMID- 9346743 TI - Liver transplantation for alcoholic liver disease: a survey of transplantation programs in the United States. PMID- 9346744 TI - Issues in selection for and outcome of liver transplantation in patients with alcoholic liver disease. PMID- 9346745 TI - Liver transplantation for end-stage alcoholic liver disease: an assessment of outcomes. PMID- 9346747 TI - Liver transplantation for alcoholic liver disease at King's College Hospital: survival and quality of life. PMID- 9346746 TI - Survival and quality of life after liver transplantation for acute alcoholic hepatitis. AB - The applicability of liver transplantation for ALD remains limited because of ethical arguments and also because of the perception of poor outcome after transplantation. Patients with alcoholic cirrhosis are known to do as well as patients with nonalcoholic liver disease after receiving liver allografts; however, the outcome in patients with severe acute alcoholic hepatitis in this setting is unclear. We studied 9 liver transplant recipients in whom severe acute alcoholic hepatitis was retrospectively diagnosed; 8 had underlying cirrhosis, and 1 had advanced fibrosis. All had Maddrey's discriminant function > 32, and most had hepatic encephalopathy and hepatorenal syndrome. History regarding abstinence was unreliable in some patients. Episodes of acute cellular rejection responded quickly to therapy, and despite recidivism in some patients, long-term survival was comparable to that of patients receiving transplants with alcoholic cirrhosis alone and those with a milder degree of alcoholic hepatitis and cirrhosis. This study suggests that severe acute alcoholic hepatitis may not be an appropriate contraindication for liver transplantation. PMID- 9346748 TI - Comorbidities of alcoholic liver disease that affect outcome of orthotopic liver transplantation. PMID- 9346749 TI - Definition and diagnosis of relapse to drinking. PMID- 9346750 TI - Long-term follow-up of patients with alcoholic liver disease who underwent hepatic transplantation. PMID- 9346751 TI - Relapse after transplantation: European studies. PMID- 9346752 TI - Predictive factors for alcoholic relapse in the selection of alcohol-dependent persons for hepatic transplant. PMID- 9346753 TI - Recurrence of alcoholic liver disease after liver transplantation. PMID- 9346754 TI - Research direction. PMID- 9346755 TI - Monitoring for alcohol use relapse after liver transplantation for alcoholic liver disease. PMID- 9346756 TI - The natural history of alcoholism and its relationship to liver transplantation. PMID- 9346757 TI - Treatment of alcohol dependence. PMID- 9346758 TI - Treatment of the postoperative alcoholic liver transplant recipient with other addictions. PMID- 9346759 TI - Current research in the treatment of alcoholism in liver transplant recipients. PMID- 9346760 TI - Transplantation for alcoholic liver disease: the ethical issues. PMID- 9346761 TI - Transplantation in alcoholics: separating prognosis and responsibility from social biases. AB - The general public does not favor transplanting livers into patients with alcoholic cirrhosis. This opinion may reflect a sense that we should not distribute scarce resources to people who are personally responsible for their illness. It may also reflect a sense that alcoholism is socially undesirable, and therefore alcoholics should not receive transplants. This article argues that these positions do not hold up under scrutiny. The only reason to give alcoholic patients lower priority for transplantation is if subgroups of alcoholics can be shown to have unacceptably poor transplant prognoses. However, giving these alcoholics lower priority is justifiable only if it is part of a larger policy that distributes livers on the basis of prognosis. In the meantime, there is no justification for giving lower priority to alcoholics for available livers. PMID- 9346762 TI - Liver transplantation for alcoholic liver disease: executive statement and recommendations. Summary of a National Institutes of Health workshop held December 6-7, 1996, Bethesda, Maryland. PMID- 9346763 TI - Portal vein complications after liver transplantation for biliary atresia. AB - The objective of this report is to review portal complications (PC) after pediatric liver transplantation (LT) for biliary atresia (BA) in the Bicetre surgical series. From January 1, 1988, to February 28, 1995, 96 children with BA underwent 115 LTs Portal anastomosis was done on either the recipient portal vein (n = 85) or superior mesenteric vein (n = 11). No antiaggregative agents were administered postoperatively. Median follow-up was 50 months (range, 12 to 97). Nineteen PC (16.5%) occurred in 17 recipients: 16 portal thrombosis (PT) and 3 portal stenosis (PS). Fifteen instances of early PT occurred between days 0 and 17 (median, day 2). Emergency thrombectomy was performed in 9 cases (successful in 5). Three children underwent a secondary portosystemic shunt (successful in 2). Three PS were cured by either surgery or balloon dilatation. Four children died, 3 are alive with portal hypertension (PHT), and 10 are alive without PHT. Three-year patient actuarial survival is 82.4% in PC cases and 82% in others (NS). Significant risk factors of PC are young age and weight at the time of LT, small portal vein, and emergency LT. Analysis of our own results and review of the literature suggest that prevention of PC depends primarily on appropriate surgical technique. Reduction of postoperative hypercoagulability may also play an important role: a meta-analysis of 1,257 published pediatric LT show an overall risk of PT of 2.2% in teams using aspirin with or without dipyridamole compared with 7.8% when no antiaggregative agents are given (P = .0001). PMID- 9346764 TI - Liver transplantation for cryptogenic cirrhosis. AB - End-stage liver disease secondary to cryptogenic cirrhosis is the indication for orthotopic liver transplantation (OLT) in 7% to 14% of recipients. However, there are no reports documenting the outcome of OLT for this indication. The aim of this study was to determine (1) survival and (2) the incidence of histological recurrence of cryptogenic cirrhosis after OLT. Between March 1985 and December 1994, 560 OLTs were performed at our institution. Of these, 39 transplants for cryptogenic cirrhosis were in patients who met the following criteria: antinuclear antibody < 1:40; negative anti-smooth muscle antibody, antimitochondrial antibody, polymerase chain reaction for hepatitis C virus, and hepatitis B surface antigen results; normal ceruloplasmin and alpha-1 antitrypsin phenotype; transferrin saturation < 65%; and liver biopsy specimen not suggestive of hemochromatosis or other known disorders. Histological recurrence was assessed with protocol liver biopsies in all patients who survived longer than 6 months. The mean age of cryptogenic recipients at the time of transplantation was significantly lower (40.6 years; range, 3 to 63 years) than that of noncryptogenic recipients (48.5 years; range, 1-70; P < .03). Median modified Child's-Pugh score was slightly higher for cryptogenic recipients at the time of transplantation (10.0 + 0.08 standard error of mean [SEM]), than for the noncryptogenic recipients (9.0 + 0.03 SEM; P < .02). Actuarial survival was 72% (+ 0.07 SEM) at 1 and 58% (+ 0.08 SEM) at 5 years for cryptogenic recipients compared with 89% at 1 and 80% at 5 years for noncryptogenic recipients. The difference in survival was significant (P < .001) at both 1 and 5 years. Among the 27 cryptogenic recipients surviving more than 6 months (mean follow-up, 5.5 years), 6 have persistent hepatitis histologically without apparent infectious, vascular, biliary, or drug origins. Four patients (15%) had chronic active hepatitis, and 2 (7%) had steatohepatitis. No cases of recurrent cryptogenic cirrhosis were seen. OLT for cryptogenic cirrhosis is associated with a poor outcome compared with other indications, hepatitis of uncertain origin occurred in 22% of cryptogenic recipients surviving longer than 6 months, and no evidence of recurrence of cryptogenic cirrhosis was seen thus far in follow-up. PMID- 9346765 TI - Biliary reconstruction without T tubes or stents in liver transplantation: report of 502 consecutive cases. AB - Although T tubes and stents are widely used as part of the routine biliary reconstruction in liver transplantation, they have inherent complications and there is no proof that they are beneficial to healing. We do not use T tubes or anastomotic stents, and we reviewed our experience with 502 consecutive, whole size liver grafts to determine the incidence and nature of biliary complications. Duct-to-duct (D-D) and Roux-en-Y loop-to-duct (RY-D) anastomoses were performed in 321 and 176 cases, respectively. In 62% of cases, the donor gallbladder was transplanted and an external catheter cholecystostomy was fashioned to provide for postoperative cholangiography. In the remaining cases the gallbladder was removed. Biliary complications of all types occurred after 13.5% of the transplants. Anastomotic complications (stricture, obstruction, or leak) occurred in 8.2% of the cases, and they were least frequent (4.0%) with RY-D reconstructions. Gallbladder-related complications accounted for one quarter of all biliary complications, and they outweighed the advantage of convenient access to the biliary tree for cholangiography. Four patients (0.9%) died of biliary complications. We conclude that routine reconstruction of the biliary tract without T tubes or stents is a safe technique in liver transplantation. Retaining the donor gallbladder as a method of providing cholanglography is not necessary. PMID- 9346766 TI - Receiver operating characteristic analysis for biliary complications in liver transplantation. AB - Receiver operating characteristic (ROC) analysis was used to assess the use of the serum chemical markers, gamma-glutamyl transferase (GGT), alkaline phosphatase (AP), and total bilirubin (BR) as tests for biliary complications in patients who had undergone orthotopic liver transplantation. Our study consisted of 105 consecutive adult transplant patients at the University of Chicago from March 1985 to November 1988. Biliary complications were determined by cholangiogram. Maximum serum values for three postoperative time periods (days 0 to 30, days 31 to 90, and > 90 days) were obtained for each patient. ROC analysis showed that GGT was the best single test during the earliest and latest time periods, whereas BR was best during days 31 through 90. We also assessed the time periods, surgical biliary anastomosis, pretransplant diagnosis, and location of biliary lesions compared with the incidence of biliary pathology for the patients. We found that patients with a pretransplant diagnosis of acute parenchymal liver disease were more likely to have biliary complications, and patients with end-to-end anastomosis with T tube were also more likely to have biliary complications. We further conclude that GGT, BR, and AP are all useful in screening for biliary complications and should be used routinely in liver transplant patients. PMID- 9346767 TI - Circulatory pathophysiology and options in hemodynamic management during adult liver transplantation. PMID- 9346768 TI - Hepatic artery reconstruction in living donor liver transplantation from the microsurgeon's point of view. AB - Microvascular surgery for the reconstruction of the hepatic artery in living donor liver transplantation is discussed from the microsurgeon's point of view. A refined operative procedure to improve the safety of the anastomosis is described. In living donor liver transplantation, the hepatic artery of the graft is short and small, the operative site is deep and mobile, and the anatomic arrangement of the graft left hepatic artery may differ from that of the recipient's dilated hepatic artery. To create a safe anastomosis under these conditions, recipient arteries that were slightly smaller than the graft artery were dissected. Without the size discrepancy, and end-to-end anastomosis could be created. Some refinements to create a good operative field made the anastomosis easy. The apparatus and techniques used in free-flap transfer facilitated a clean anastomosis. We anastomosed 44 arteries in 40 patients undergoing living donor liver transplantation using microsurgical techniques. Neither a decrease in the arterial blood flow nor hepatic artery thrombosis was noted. The refined operative procedure we describe in this report can be used to overcome the problems associated with the hepatic artery anastomosis in living donor liver transplantation. PMID- 9346769 TI - Loss of serum HBsAg after interferon-A therapy in liver transplant patients with recurrent hepatitis-B infection. AB - Reinfection with hepatitis B virus after orthotopic liver transplantation is nearly universal in patients who have not received posttransplant immunoprophylaxis. Recurrence almost invariably leads to chronic liver disease. Interferon has been used both prophylactically and therapeutically but has not been effective. We treated 2 liver transplant patients with recurrent hepatitis B virus (HBV) infection (serum hepatitis B surface antigen [HBsAg] and HBV DNA positive on polymerase chain reaction, and positive liver biopsy result) with interferon, 3 to 6 MU three times weekly for 6 to 22 months. A full response to therapy was manifested in both patients by normalized serum alanine aminotransferase levels and the loss of serum HBsAg and HBV DNA. The effectiveness of interferon in our patients may have been related to coinfection with hepatitis D virus in the first case and the high interferon dose (6 MU, three times weekly) and long treatment period (22 months) in the second. No episodes of rejection were noted during therapy. We conclude that interferon can induce a complete response in liver transplant patients with recurrent HBV infection. Future studies should investigate the use of interferon therapy at higher doses and/or for longer periods. PMID- 9346770 TI - Comparison of histopathology in acute allograft rejection and recurrent hepatitis C infection after liver transplantation. AB - Recurrent hepatitis C infection after orthotopic liver transplantation (OLT) is frequent and may occur as early as a few weeks postoperatively. Early histopathological features of recurrent hepatitis C virus (HCV) infection may be modified by immunosuppressive therapy and can be difficult to differentiate from acute allograft rejection (AAR). Thus, we retrospectively compared histopathological features of liver biopsy specimens from two carefully selected patient groups: one with unequivocal recurrent hepatitis C, the other with unequivocal AAR. Index biopsy specimens obtained at the time of the appearance of liver test abnormalities after OLT and all serial liver biopsy specimens (2 to 13 per patient) were assessed under code and scored semiquantitatively for 44 histopathological variables. The index biopsy specimens from patients with recurrent HCV infection and AAR index biopsies (AAR-Ib) differed significantly (P < .05) for 11 features (10 features were statistically associated with AAR and 1 with early recurrence of HCV infection). Statistically significant features associated with AAR included bile duct injury with overlapping nuclei, lymphocytic infiltrates and necrosis, endothelialitis, portal inflammatory infiltrates containing eosinophils and polymorphonuclear leukocytes, hepatocyte mitoses, and zone 3 canalicular cholestasis. In contrast, the only statistically significant feature associated with early recurrent HCV was sinusoidal dilatation. Stepwise discriminant analysis showed that the presence of eosinophils in the portal inflammatory infiltrate, bile duct necrosis, and bile duct lymphocytic infiltrates were independently associated with AAR. However, serial biopsy specimens from patients with recurrent HCV infection showed statistically significant progression in scores for portal inflammation, portal lymphoid aggregates, and lobular inflammation. We conclude that (1) multiple histopathological features are associated with AAR; (2) early recurrent HCV infection is characterized by elevated alanine aminotransferase levels, positive HCV RNA by polymerase chain reaction (PCR), and absence of diagnostic histopathological features; and (3) serial biopsies are needed to demonstrate progression of histopathological features of recurrent hepatitis C. PMID- 9346772 TI - Hyperlipidemia in liver transplant recipients: prevalence and risk factors. AB - Hyperlipidemia is common in transplant patients. Although a causal relationship to the use of cyclosporine is accepted, additional risk factors are as yet unidentified. Eighty-five liver transplant recipients treated with standard triple immunosuppression with a survival of at least 6 months were evaluated. Pretransplantation and posttransplantation variables were analyzed as predictive factors of posttransplantation hyperlipidemia. Serum cholesterol and triglyceride levels were considered elevated if they were > 250 mg/dL and > 150 mg/dL, respectively. Before and after transplantation, hyperlipidemia occurred in 8% (95% confidence interval [CI], 3% to 16%) and 66% (95% CI, 55% to 76%), respectively. After transplantation, 47% (95% CI, 36% to 58%) of the patients had isolated high triglyceride levels, 12% (95% CI, 6% to 21%) had both elevated cholesterol and triglyceride levels, and 7% (95% CI, 3% to 15%) had isolated elevated cholesterol levels. Hypertriglyceridemia occurred early after transplantation (67% by first month) and persisted nearly unchanged throughout the first year. In contrast, cholesterol levels increased with time (5%, 13%, and 27% at 1, 3, and 6 months, respectively). In univariate analysis, factors predictive of hypercholesterolemia included female sex, pretransplantation cholestatic liver disease, pretransplantation cholesterol levels > 141 mg/dL, and > 3 methylprednisolone "boluses." In multivariate analysis, only a pretransplantation cholesterol level of > 141 mg/dL (odds ratio [OR], 5.5; 95% CI, 1.4 to 21) was an independent risk factor. Risk factors associated with hypertriglyceridemia included pretransplantation hepatocellular liver disease (OR, 6.8; 95% CI, 1.2 to 40) and posttransplantation renal dysfunction (OR, 5.4; 95% CI, 1.9 to 15.4). Hyperlipidemia is a frequent finding in liver transplant recipients, and hypertriglyceridemia is the most common abnormality. Hypertriglyceridemia can be predicted on the basis of pretransplant hepatocellular disease and posttransplant renal dysfunction. Pretransplant serum cholesterol level is an independent risk factor for posttransplant hypercholesterolemia. PMID- 9346771 TI - Apoptosis after ischemia-reperfusion in human liver allografts. AB - Little is known about the possible contribution of apoptosis to ischemia reperfusion injury in human liver transplantation. Therefore, we studied postreperfusion surgical biopsy specimens of 16 human liver allografts using the TUNEL assay for in situ demonstration of apoptotic cells. In all patients, a variable proportion of hepatocytes and sinusoidal endothelial cells presented labeled nuclei. The mean +/- standard deviation percentages of positive hepatocytes were 18.7% +/- 12.2% in the whole section, 30.4% +/- 18.7% in the subcapsular region, 14.5% +/- 13.5% in the centrilobular zones, and 10.3% +/- 9.5% in the periportal zones. The percentage of positive hepatocytes were not correlated with the duration of cold ischemia but was higher in grafts harvested from donors with elevated preoperative aspartate aminotransferase (AST) levels. The percentage of positive hepatocytes was correlated with postoperative serum levels of AST (P = .015) and inversely correlated with postoperative serum levels of factor V (P = .019). Apoptotic biliary epithelial cells were detected in only 3 cases. In conclusion, apoptosis is a frequent event in postreperfusion biopsy specimens of liver allografts and probably contributes to preservation injury of hepatocytes. PMID- 9346773 TI - Reassessing the role of medical therapy in the management of hepatic vein thrombosis. AB - Hepatic venous outflow obstruction caused by hepatic vein thrombosis (HVT) is a manifestation of a hypercoagulable state, most commonly a myeloproliferative disorder (MPD). In the past, HVT was thought to have a poor prognosis unless treated surgically with portosystemic shunt or orthotopic liver transplantation (OLT). The aim of this study was to assess whether early diagnosis of the underlying hematologic disorder and institution of appropriate medical therapy have altered outcome. We reviewed the charts of 22 patients with HVT evaluated at our center from January 1986 to January 1995. The median age was 32 years (range, 14 to 59 years). Underlying etiologies were MPD, 13 (polycythemia vera, 8; essential thrombocythemia, 4; undefined, 1); dysfibrinogenemia, 1; anticardiolipin antibody, 1; oral contraceptive use, 3; and idiopathic, 4. All patients had ascites, hepatomegaly, and/or abdominal pain. Two underwent mesocaval shunting, and 1 had a peritoneal-venous shunt. Seven patients, including 1 with a mesocaval shunt, underwent OLT. The median duration of symptoms before transplantation was 6 months (range, 1.5 to 11 months). Six transplant patients are alive on long-term anticoagulation therapy at a mean post OLT follow-up of 42 months (range, 2 to 77 months), without recurrence. Of 13 patients treated medically, 10 (77%) are alive at a median follow-up of 40 months (range, 17 months to 14 years 8 months), 1 has died, and 2 have been lost to follow-up. In a majority of patients, symptoms improve with prompt treatment of the underlying hematologic disorder, with a favorable long-term prognosis. Patients with decompensated liver disease can successfully undergo OLT with a low risk of recurrence on long-term oral anticoagulation. PMID- 9346774 TI - Acute pancreatitis after orthotopic liver transplantation in children. PMID- 9346775 TI - Disseminated histoplasmosis after orthotopic liver transplantation. PMID- 9346776 TI - Staphylococcal scalded skin syndrome in a liver transplant patient. PMID- 9346777 TI - Progress in posttransplant hyperlipidemia. PMID- 9346778 TI - Budd-Chiari syndrome: decisions, decisions. PMID- 9346779 TI - Fast tracking in liver transplantation. PMID- 9346780 TI - Practice patterns and anesthesia-related costs for liver transplantation. PMID- 9346781 TI - Evaluation of patients with portal hypertension-associated pulmonary artery hypertension. PMID- 9346782 TI - Intraoperative management of liver transplant patients with pulmonary hypertension. PMID- 9346783 TI - Infection control issues and intraoperative care. PMID- 9346784 TI - Monitoring and handling of reperfusion. PMID- 9346785 TI - How to do 2 a.m. research. PMID- 9346786 TI - Coagulation and liver transplantation: current concepts. PMID- 9346787 TI - The evolution of a successful liver transplant program in 1996: the clinical and administrative role of the anesthesiologist. AB - The establishment of a new liver transplant program requires enormous planning and resources. Extensive negotiations must take place to ensure institutional and departmental commitments to obtain the proper equipment, personnel, and other resources. The formation of a well-trained multidisciplinary team of physicians and nurses becomes the next step. Finally, ample time must be provided to adequately deploy resources, lobby referring physicians, recruit patients, and troubleshoot problems as they arise. PMID- 9346788 TI - Issues in setting up new liver transplant programs: transplant anesthesia in a community hospital. PMID- 9346789 TI - Selective shunts for portal hypertension: current role of a 21-year experience. AB - The results of treatment of hemorrhagic portal hypertension with selective shunts over a 21-year period in a selected patient population are reported. Patients selected for surgical treatment had good cardiopulmonary and renal function, and most also had adequate liver function (141 Child-Pugh class A, 59 class B). Among 734 patients treated surgically for bleeding portal hypertension, 221 had selective shunts (168 distal splenorenal and 53 splenocaval shunts). Global operative mortality (in the 21-year period) was 14% and 12% for Child-Pugh A patients. Operative mortality in Child-Pugh A patients in the last 5 years was only 5%. The rate of rebleeding was 6%, rate of incapacitating encephalopathy was 5%, and rate of survival was 65% at 15 years (last 5 years: 88% at 1 year and 85% at 5 years). Good quality of life was demonstrated in 80% of surviving patients. Shunt patency was 94%. Postoperative portal blood flow changes occurred in 23% of cases (8% diameter reduction, 14% thrombosis). Compared with other forms of therapy (pharmacotherapy, sclerotherapy, and transjugular intrahepatic shunting), only liver transplantation offers similar results for these patients. In countries in which liver transplantation is not routinely performed, shunting with selective shunts is the treatment of choice for patients with good liver function. PMID- 9346790 TI - Liver transplantation: current and potential applications of magnetic resonance spectroscopy. AB - Magnetic resonance spectroscopy (MRS) allows the noninvasive measurement of whole organ metabolism due to the presence of the MR-sensitive nucleus phosphorus 31 in adenosine triphosphate (ATP), its precursors, and break-down products. In small animal liver transplant studies it has been used to analyze the metabolic effects of cold and warm ischemia, hypothermic reperfusion, and the relative efficacy of different organ preservation solutions. In recent large animal studies MRS has been developed to provide continuous dynamic information on ATP metabolism during graft reperfusion and the bioenergetic consequences of altering preservation solutions. These basic experimental data need to be critically evaluated in human liver transplantation. Encouraging preliminary data on many possible clinical applications have already been obtained, such as the assessment of human donor liver viability and posttransplant graft function. At present, the cost and technically demanding nature of MRS may restrict its application to research units. PMID- 9346791 TI - Severe pulmonary hypertension in liver transplant candidates. AB - Advanced liver disease with portal hypertension may be associated with pulmonary hypertension. A review of 1,205 consecutive liver transplant patients was made to assess the incidence and severity of pulmonary hypertension in patients with end stage liver disease. Postoperative data were reviewed to determine if outcome was influenced and, in patients with severe pulmonary hypertension, whether pulmonary hypertension was reversed after transplantation. The hemodynamic data of 5 patients who were found to have severe pulmonary hypertension before transplantation and did not receive transplants were also reviewed. The incidence of pulmonary hypertension in the patients who received transplants was 8.5% (n = 102; mean pulmonary artery pressure, > 25 mmHg). The incidence of mild pulmonary hypertension was 6.72% (n = 81; systolic pulmonary artery pressure, 30 to 44 mmHg); that of moderate pulmonary hypertension was 1.16% (n = 14; systolic pulmonary artery pressure, 45 to 59 mmHg); and that of severe pulmonary hypertension was 0.58% (n = 7; systolic pulmonary artery pressure, > 60 mmHg). Mild and moderate pulmonary hypertension did not influence the outcome of the procedure. Severe pulmonary hypertension was associated with mortality rates of 42% at 9 months posttransplantation and 71% at 36 months posttransplantation. Only 2 of 7 patients with severe pulmonary hypertension have survived liver transplantation with a good quality of life. The remaining 5 patients continued to deteriorate with progressive right heart failure with no evidence of amelioration of the pulmonary hypertension. This experience supports the view that in most patients who have severe pulmonary hypertension associated with advanced liver disease, it is caused by fixed pathological changes in the pulmonary vasculature, is not reversible with liver transplantation, and is associated with a very high perioperative mortality rate. PMID- 9346792 TI - Hepatitis C virus genotypes and quantitation of serum hepatitis C virus RNA in liver transplant recipients: relationship with severity of histological recurrence and implications in the pathogenesis of HCV infection. AB - The reasons for the wide variation of incidence and severity of recurrent hepatitis C after liver transplantation are not clear. We have studied liver transplant recipients to assess the impact of hepatitis C virus (HCV) genotype and HCV RNA quantification on HCV recurrence after transplantation. Twenty-two patients received transplants for HCV cirrhosis and were followed up with virological and histological assessments. Mean follow-up was 39 months. HCV genotype was determined with line probe assay (Inno-Lipa). HCV RNA quantity was determined in serum samples by use of polymerase chain reaction nested assay. HCV genotype 1 was detected in 13 patients and other genotypes in 9. Histological recurrence rates were 69% in patients with genotype 1 and 66% in patients with other genotypes. All cases of severe histological injury (chronic active hepatitis or cirrhosis) were observed in patients with genotype 1. HCV RNA quantity was significantly higher in patients with genotype 1 (mean, 2.023 x 10(3) copies/mL) than in patients with other genotypes (mean, 27,403 copies/mL). In conclusion, the severity of histological recurrence after liver transplantation for HCV disease was higher in patients infected by HCV genotype 1 than in those infected with other genotypes. The levels of viral replication were higher in patients with HCV genotype 1 than in those with other genotypes. PMID- 9346794 TI - Cost analysis of intraoperative blood salvage during orthotopic liver transplantation. AB - Approximately 6,000 to 7,000 orthotopic liver transplantation (OLT) procedures are performed annually, which require the administration of large volumes of blood products. Thus liver transplantation can significantly strain local and regional blood resources at a time when transfusion practices are changing dramatically, in large part because of anxiety caused by the human immunodeficiency virus. Intraoperative autologous transfusion has been proposed as a means of both reducing transfusion demands and lessening the hazards of allogeneic transfusion. However, the cost effectiveness of intraoperative blood salvage has not been unequivocally determined. We retrospectively examined the cost of intraoperative autologous transfusion during OLT for a 2-year period at the University of Cincinnati Hospital. A direct comparison was made between the charge for autologous transfusion and the calculated cost of allogeneic transfusion. Seventy OLT procedures were performed during the years 1993-1994. The average charge for autologous transfusion was $1,048.73 per case. Cell salvage volumes for all cases were added, and the calculated conservation of allogeneic packed red blood cells totaled 359.6 units, worth $30,026.60 or $428.95 per case. The break-even point is approximately 12.6 units, and most patient do not receive this volume of salvaged blood. In fact, cell salvage reached cost equivalence in only three cases (4.8%). Moreover, the cost deficit of autologous transfusion during this 2-year period averaged $586.56 per case. PMID- 9346793 TI - Enhanced (cytomegalovirus) viral replication after transplantation for fulminant hepatic failure. AB - Fulminant hepatic failure (FHF) is an established indication for liver transplantation. A pretransplant diagnosis of FHF may be a risk factor for subsequent development of cytomegalovirus (CMV) infection, although the mechanism of this association is not understood. FHF is associated with very high levels of tumor necrosis factor alpha (TNF-alpha), and TNF-alpha may directly promote viral replication. We have used the polymerase chain reaction (PCR) to examine sequentially collected buffy coats from 106 consecutive adult liver transplant recipients. PCR evidence of CMV replication was found for 13 of 18 patients who underwent transplantation for FHF (c.f. 23/88 non-FHF patients; P < .01). Ten of 12 patients who received transplants transplanted for fulminant seronegative hepatitis were buffy-coat PCR-positive, sometimes during the first posttransplant week. TNF-alpha was measured by enzyme-linked immunosorbent assay in selected sera, and results were examined in the context of a quantitative PCR assay. Serum TNF-alpha levels increased and decreased in concert with viral titers. High levels of TNF-alpha were not found in the early posttransplant period. We conclude that FHF (in particular, seronegative hepatitis) is associated with enhanced CMV replication in the posttransplant period. Results also suggest that viral replication may be enhanced before transplantation. These patients may be at special risk for development of symptomatic CMV infection. PMID- 9346795 TI - Orthotopic liver transplantation with preservation of portocaval flow compared with venovenous bypass. AB - Conventional liver transplantation requires cross-clamping of the hepatic pedicle and inferior vena cava, leading to severe hemodynamic and metabolic disturbances, usually attenuated by the use of venovenous bypass. A more recent surgical technique, piggyback with temporary portocaval shunting, preserves both caval and portal blood flows. The aim of this study was to compare the two methods prospectively. Forty-four patients with chronic liver disease were studied. Local anatomic conditions guided the surgeon to choose the easiest way to remove the native liver. Anesthetic management was standardized. Hemodynamic and metabolic changes were assessed by use of routine tests at specific periods. Graft function was evaluated by measurement of aminotransferases and monoethylglycinexylidide (MEGX) test 12, 24, 48, and 72 hours postoperatively. Conventional liver transplantation with venovenous bypass was performed in 26 patients, and the piggyback with temporary portocaval shunting was performed in 15 patients. ANOVA showed that cardiac output and systemic oxygen delivery were better maintained before revascularization in the piggyback group. Metabolic changes were comparable, and hyperfibrinolytic activity was detected in both groups. Graft function was comparable and satisfactory within the 3 first postoperative days. Piggyback with temporary portocaval shunting provided better intraoperative hemodynamics and tissue oxygenation than liver transplantation with venovenous bypass. PMID- 9346796 TI - Acute cyclosporine toxicity after liver transplantation is predicted by the lidocaine monoethylglycinexylidide test in the donor. AB - Cyclosporine toxicity is still a significant problem in the early period after liver transplantation. The monoethylglycinexylidide (MEGX) test performed in the donor has been suggested as a reliable test to predict liver graft function in the recipient. The MEGX test was performed in 50 consecutive donors, and the clinical course of recipients, metabolic parameters of the grafts, and cyclosporine levels were followed in detail for 10 days. Two patients died of sepsis and were excluded. Renal and/or neurological toxicity appeared in 15 of the remaining 48 patients (31%). In the 6 with neurological problems, MEGX values were low (41, 47, 50, 60, 94, and 101 micrograms/L). Nine patients had transient elevations of creatinine and urea; in 8 of these, cyclosporine levels remained in the normal range. Low MEGX values in the donors correlated with early evidence of cyclosporine toxicity (P < .0001), reduced graft function (bile output, P = .04; prothrombin time at day 5, P = .005), and prolonged stay in the intensive care (P = .022). The MEGX test is valuable and reflects the metabolic capacity of liver grafts. It can predict posttransplantation complications caused by cyclosporine toxicity, and further study would evaluate its incorporation into immunosuppressive protocols. PMID- 9346797 TI - Lack of relationship between preoperative measures of the severity of cirrhosis and short-term survival after liver transplantation. AB - The purpose of this study was to evaluate the prognostic value of clinical measures of the severity of disease in cirrhotic patients who were candidates for liver transplantation at our institution. The records of the 132 cirrhotic patients who were candidates for a first transplantation between January 1, 1987, and December 31, 1994, were reviewed. One hundred nine patients (82.6%) received grafts, and 23 (17.4%) died while on the waiting list. The variables examined included level of medical urgency at the time of enlistment, date of transplantation, serum creatinine level, variables that constitute the Child-Pugh score and Shaw's risk score (serum bilirubin and albumin, prothrombin time, ascites, encephalopathy, nutritional status, age, and operative blood loss), and 6-month survival status after transplantation. The proportion of patients who died awaiting a graft increased as a function of the Child-Pugh score at enlistment (score 5-6, 0%, n = 6; score 7-9, 7%, n = 54; score 10-11, 18%, n = 33; score 12-15, 33%, n = 39; P = .01). Six-month survival rates after transplantation were similar irrespective of the Child-Pugh score or Shaw's risk score. Stepwise multiple logistic regression models identified the degree of ascites, serum bilirubin, and operative blood loss as significant variables for the prediction of overall mortality 6 months posttransplantation (model chi 2 = 12.8; P = .025; r = 0.32), but the model explained only 10% of the outcomes observed. We concluded that the Child-Pugh score is a valid prognostic index for survival up to the time of transplantation for cirrhotic patients on the waiting list; however, clinical measures of the severity of cirrhosis are poor predictors of 6-month survival after transplantation. PMID- 9346798 TI - Reduced cyclosporine absorption preceded acute allograft rejection in a child with a liver transplant. AB - This case report correlates impaired cyclosporine absorption from the traditional oral formulation in a 9-year-old liver transplant recipient with subsequent acute allograft rejection. Although impaired absorption in this patient was documented by cyclosporine pharmacokinetic profiling (steady-state area under the cyclosporine concentration-time curve or AUC), no indication was evident from the pre-dose cyclosporine trough level, which was within the typical target range of blood concentrations. However, when the subject received the microemulsion formulation of cyclosporine the AUC value reflected an adequate absorption pattern. We recommend that if malabsorption is suspected in the de novo pediatric liver transplant patient, then single-sample pre-dose trough cyclosporine levels should not be relied on as an indicator of sufficient immunosuppression and that a limited sampling strategy be used to confirm or rule out impaired absorption. PMID- 9346799 TI - Kaposi's sarcoma presenting as a protracted multisystem illness in an adolescent liver transplant recipient. AB - Kaposi's sarcoma (KS) is a common malignancy in patients with acquired immunodeficiency syndrome (AIDS), classically appearing as red to purple plaques containing small papules and nodules. We report our experience with an adolescent orthotopic liver transplant recipient who presented with an unusual presentation of KS. The patient had a protracted multisystem illness that began with hemolytic anemia, fevers, and fatigue and progressed to pancreatitis, sinusitis, lymphadenopathy, and mouth ulcers. The diagnosis was made by a lymph node biopsy that was performed to evaluate for Epstein-Barr virus. The classical subcutaneous nodules characteristic of KS did not become evident until shortly before the patient died. We present this case to emphasize that KS in pediatric liver transplant patients can present as a multisystem disease that progresses to disseminated organ involvement before the characteristic subcutaneous manifestations are evident. PMID- 9346801 TI - Selective shunts in the 1990s. PMID- 9346800 TI - Cytomegalovirus infection, fulminant hepatitis, and liver transplantation: the sides of the triangle. PMID- 9346802 TI - Severe exacerbation of chronic hepatitis B and ensuing viral mutation after reduction of immunosuppression in a liver transplant recipient. PMID- 9346818 TI - Molecular dynamics of bacteriorhodopsin. AB - A model of bacteriorhodopsin (bR), with a retinal chromophore attached, has been derived for a molecular dynamics simulation. A method for determining atomic coordinates of several ill-defined strands was developed using a structure prediction algorithm based on a sequential Kalman filter technique. The completed structure was minimized using the GROMOS force field. The structure was then heated to 293 K and run for 500 ps at constant temperature. A comparison with the energy-minimized structure showed a slow increase in the all-atom RMS deviation over the first 200 ps, leveling off to approximately 2.4 A relative to the starting structure. The final structure yielded a backbone-atom RMS deviation from the crystallographic structure of 2.8 A. The residue neighbors of the chromophore atoms were followed as a function of time. The set of persistent near residue neighbors supports the theory that differences in pKa values control access to the Schiff base proton, rather than formation of a counterion complex. PMID- 9346819 TI - Applications of nuclear magnetic resonance spectroscopy and molecular modeling to the study of protein-carbohydrate interactions. AB - This work provides an overview of the applications of NMR to the study of protein carbohydrate interactions. The use of TR-NOE experiments in this context is given. In particular, the study of Ricin/lactose and Hevein/chitobiose complexes is detailed. PMID- 9346820 TI - Conformational analysis using distance geometry methods. AB - Distance geometry methods have been used extensively to build models of molecules of various sizes, including small molecules, peptides, and proteins. These methods are often overlooked as tools for conformational analysis, even though they often perform as well as other conformational sampling methods. We have implemented two new distance geometry approaches in the DGEOM95 package. In the first new method, the traditional embedding algorithm is replaced with a procedure that generates random 4D coordinates for each atom, followed by refinement of these coordinates into 3D using the distance geometry error function. The conformational sampling produced by this method is comparable to that obtained with partial metrization, and superior to that obtained with the original embedding procedure. In the second method, a molecular dynamics step is included in the refinement stage. Although this method can be applied to any embedding algorithm, substantial improvements in sampling are seen primarily with the original embedding algorithm. PMID- 9346821 TI - Conformational analysis of biantennary glycans and molecular modeling of their complexes with lentil lectin. AB - Some mannose-binding legume lectins show higher affinity for fucosylated glycans than for glycans without fucose. These lectins possess a secondary binding site. Owing to the possibility of additional fucose binding, oligosaccharides adopt different conformations depending on whether they contain fucose or not. To study these conformational differences, complexes of fucosylated and unfucosylated glycans with Lens culinaris lectin have been modeled. Starting points were X-ray structures of lentil lectin and complexes of the homologous Lathyrus ochrus lectin. The SYBYL molecular modeling package with the TRIPOS force field was used. Two different models were built, displaying in both a network of hydrogen bonds between the saccharide and the binding site. Furthermore, to compare the free and bound ligand, conformational analysis in the free state has been performed. A complete analysis of all possible disaccharide fragments has been performed using the MM3 force field. A CICADA analysis employing the same force field was carried out to study the complete oligosaccharide. Low-energy conformers found by CICADA were clustered in conformational families and analyzed in terms of flexibility and rotational barriers. All values of glycosidic torsion angles are in the range as calculated by MM3 for the disaccharides. PMID- 9346822 TI - Drug-motif-based diverse monomer selection: method and application in combinatorial chemistry. AB - This article describes a strategy to explore monomer diversity while incorporating drug motif knowledge into the design and selection of monomers for combinatorial chemistry. The process involves collecting from available electronic databases all those molecules that potentially could be monomers. In this manner we have assembled five DAYLIGHT databases, each containing one of the common functional groups: carboxylic acids, aldehydes, nitriles, primary amines, and secondary amines. The molecules in the databases are then subjected to fingerprint and cluster analysis using the Jarvis-Patrick algorithm and profiles of the compounds are calculated relating to molecular weight, H-bond counts, and rotatable bond flexibility. The cluster information and profiles of the molecules are stored back into the databases for similarity and diversity searches, and for profile prescreening of monomers. To apply drug motif knowledge to a selection an application to an aldehyde set is discussed, in which representatives of each cluster in the aldehyde database are compared with drug molecules in the Standard Derwent File (SDF) in one of three ways to select drug motif-based monomers for purchase or synthesis. PMID- 9346823 TI - Synthesis and characterization of human gene 1 relaxin peptides. AB - The peptide encoded for by one of the two relaxin genes found in the human genome, designated H1, has been synthesized by the Boc-polystyrene solid phase method. The two chains which constitute relaxin, A- and B-, were assembled separately and, after cleavage, deprotection and purification, combined in solution at high pH to form the one intra- and two intermolecular disulfide bonds. Comprehensive chemical characterization including ion spray mass spectrometry of the peptide confirmed both its correct identity and high purity. The synthetic H1 relaxin was analyzed by circular dichroism spectroscopy and shown to possess a greater alpha-helical conformation in water than the corresponding H2 relaxin. The peptide had powerful direct chronotropic and inotropic effects in the isolated rat heart assay as did an analogue of the peptide in which the C-terminus of the B-chain was extended by four residues. PMID- 9346824 TI - Synthesis of GnRH analogs having direct antitumor and low LH-releasing activity. AB - New chicken I GnRH agonists and antagonists have been synthesized and tested for their biological activities. The common feature of these analogs was that the molecules had a beta-L-aspartyl residue inserted in position 6. The agonist bound to the pituitary still had low endocrinological activity. On the other hand, it exhibited direct antitumor effect in in vitro assays. The endocrinological activity of the antagonist was low; however, it showed potent, direct antitumor activity. These observations might lead to the development of new GnRH analogs with selective antitumor effect. PMID- 9346825 TI - Recognition of pituitary adenylate cyclase-activating polypeptide/vasoactive intestinal polypeptide (PACAP/VIP) hybrids and related peptides by rat brain membranes. AB - The binding to [125I]PACAP27 and adenylate cyclase activity have been investigated using rat brain membranes with substituted analogues of PACAP and VIP, including their hybrid peptides. Binding of [125I]PACAP27 was rapid, specific and reversible. Scatchard analysis revealed a single class of binding site, with a Kd = 457 +/- 117 pM, and a Bmax = 2.63 +/- 0.24 pmol.mg protein-1. Hybrids of PACAP, in which specific residues were substituted with the corresponding residues of VIP, and vice versa, as well as related analogues, were then tested for binding and adenylate cyclase activity. The results showed that N terminal residues were important for recognition. In particular, multiple substituted analogues of PACAP by VIP, and vice versa, demonstrated that positions 4, 5 and 9 play a dominant role in the recognition of PACAP Type I receptor in rat brain membranes and account for the differences observed between PACAP and VIP. Substitutions in the C-terminal region at positions 24, 25 and 26 are not crucial for recognition specificity. PACAP-analogues provide evidence that positions 1 and 6 are essential for receptor recognition. The flexibility at position 21 also appears to play a role as substitution with Ala or Phe is tolerated, while Pro shows a significant loss both in binding affinity and adenylate cyclase activity. PMID- 9346826 TI - Affinity purification of a correctly folded fragment of synthetic HIV-1 mRNA using a HIV-1 Rev peptide-ligand. AB - Formation of a macromolecular complex between the RNA binding protein HIV-1 Rev and HIV-1 mRNA is an essential prerequisite for nuclear export and subsequent expression of HIV-1 mRNA. The arginine rich peptide TRQARRNRRRRWRARQR, corresponding to residues 34-50 of HIV-1 Rev, contains the mRNA binding motif. We prepared a thioether linked Rev34-50-cellulose conjugate to affinity purify a fragment of synthetic mRNA corresponding to the high affinity binding site for Rev. The correctly folded fraction of mRNA (27.5%) was isolated from a crude synthetic mixture. PMID- 9346827 TI - 1H NMR structural study of free and template-linked antigenic peptide representing the C-terminal region of the heavy chain of influenza virus hemagglutinin. AB - A 12 kDa template-assembled molecule, incorporating four oxime-linked synthetic peptides representing residues 306-328 of influenza virus hemagglutinin (HA), has been analysed by 1H NMR spectroscopy. The molecule (referred to as the 'tetraoxime') is of interest because it has been shown to elicit a better immune response than the free, monomeric peptide not only in the production of antibodies crossreactive with HA but also in its ability to elicit CD4+ T helper cells. We describe here an NMR structural analysis of (i) the unlinked template molecule and (ii) the free peptide and show that their conformations are not affected upon assembly of the tetraoxime. Our results suggest that the increased immune response observed for the tetraoxime may be due to its greater size and valency compared to that of the free peptide rather than being due to any induced structural effects. PMID- 9346828 TI - 1H NMR studies of the effects of glycosylation on the C-terminal pentapeptide of peptide T. AB - The C-terminal pentapeptide of peptide T (T5) and a glycosylated analogue (T5GlcNAc) were investigated using 1H NMR spectroscopy to examine the influence of the sugar on the secondary structural characteristics of the peptide. The NMR data confirm the presence of a turn structure amongst an ensemble of predominantly randomly structured species in a solution of 83% TFE/H2O for both peptides. This is in agreement with a previous CD analysis demonstrating the presence of beta-turn. Unlike the CD study, the NMR data do not show a difference in the time-averaged conformation of the glycosylated versus non-glycosylated peptide. These studies suggest that any sugar-peptide interactions which occur in this system are transient in nature, and that they do not greatly influence the local secondary structural characteristics of the peptide. In particular, the turn predisposition already exhibited by the peptide appears to be neither enhanced nor reduced by a neighbouring natural N-glycosylation site. This finding is likely to be of general interest, given the importance of glycosylation as a post-translational modification and that its role in determining protein structure has yet to be characterized. PMID- 9346829 TI - Synthesis of oligonucleotides labelled with 2,4-dinitrophenyl groups at thymidine sites. AB - A phosphoramidite has been produced for labelling oligonucleotides with DNP groups at thymidine sites during solid-phase synthesis. The dinitrophenylamino group is attached via a caproamidopropargyl group to the 5-position of uracil. A related DNP-labelling phosphoramidite has been synthesised where the propargyl group is replaced by propyl. Both phosphoramidites have been used to synthesises DNP-labelled oligonucleotides. A related DNP-labelled deoxyuridine triphosphate has also been synthesised. DNP labelled oligonucleotide probes are valuable in diagnostic applications for the antibody-based detection of DNA and RNA. PMID- 9346830 TI - Antibody-mediated detection and physical properties of oligonucleotides labelled with multiple internal and terminal 2,4-dinitrophenyl groups. AB - DNP-labelled phosphoramidites have been used to synthesis oligonucleotides with multiple DNP reporter groups. The antibody-mediated detection and the stability of duplexes formed by these labelled oligonucleotides have been studied. A DNP labelled deoxyuridine triphosphate has also been used to enzymatically incorporate DNP-labels into DNA via the polymerase chain reaction. The use of DNP labelled primers in the PCR has also been investigated. PMID- 9346831 TI - Synthesis of azaalanine peptides using the solid phase method. AB - The methodology for the incorporation of azaamino-acid residues into peptides synthesised by a solid-phase method has been extended to allow azaalanine peptides to be prepared. In this way, Ac-Leu-Ser-Gly-azaAla-Gly-Phe-Ser-Leu-NH2 H Ala-Ala-Lys-Glu-Ala-Ala-Glu-Ala -Ala-Glu-Lys-Ala-azaAla-Glu-Leu-Ala-Leu-N2H3, and H-Ala-azaAla-Lys-Glu-Ala-Ala-Glu-Ala-Ala-Glu-Lys-Ala-Ala-Glu-Leu-A la-Leu-N2H3 have been prepared. A new analogue of the Ala-Gly sequence, 3-azaalanylpropionic acid has been prepared and used to prepare the peptide analogue acetyl-Leu-Ser Gly-azaAla-3-Prop-Phe-Ser-Leu-NH2. PMID- 9346832 TI - Immunohistochemical comparison of localization of pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) in the enteric nerve plexus of the guinea pig jejunum. AB - The localization and co-localization of pituitary adenylate cyclase activating polypeptides (PACAPs) and vasoactive intestinal polypeptide (VIP) in the enteric nerve plexus of the guinea pig jejunum were immunohistochemically compared by the peroxidase anti-peroxidase (PAP) method using region-specific antisera against PACAP38, PACAP27 and VIP, respectively. Immunoreactive nerve elements were demonstrated in the ganglia of both myenteric and submucous plexi in the guinea pig jejunum. Numerous immunoreactive nerve cell bodies were localized in the myenteric ganglia, while nerve cell bodies in the submucous ganglia were only slightly immunopositive. Immunoreactive nerve terminal varicosities occurred in both myenteric and submucous ganglia. Additionally, PACAPs-immunoreactive nerve cell bodies were also immunopositive with VIP. Thus, both PACAP38 and PACAP27, as well as VIP, are co-localized in the cell bodies of guinea pig jejunum. PMID- 9346833 TI - The stability and conformation of duplex DNA containing the novel anti-HIV drug ADRT. AB - The anti-HIV drug 4'-azidothymidine (ADRT) has been incorporated into DNA by the phosphoramidite method. The presence of the modified nucleotide was shown to have a minimal effect on duplex conformation and stability by CD spectroscopy and UV melting. PMID- 9346834 TI - Alteration of immunogenicity and antibody recognition of B-cell epitopes by synthetic branched chain polypeptide carriers with poly[L-lysine] backbone. AB - In order to elucidate structural and biological properties required for an optimal immunological carrier function and to provide a rational basis for its selection, two new groups of synthetic branched polypeptides with a general formula poly[Lys-(X(i)-DL-Ala(m))][XAK] or poly[Lys-(DL-Ala(m)-X(i))][AXK], where m approximately 3 and i < 1 were introduced by our laboratory. Here we review our recent results on the application of these polypeptides as biodegradable carriers for constructing synthetic immunogens/antigens with a well-known phenyl oxazolone hapten, peptide epitopes of epithelial mucin [MUCI] or herpes simplex virus [HSV 1] glycoprotein D. Observations collected during the last five years with the conjugates presented serve to illustrate the usefulness of branched polypeptides as carriers for the rational design of synthetic immunogens for the development of vaccines or clinically relevant immunodiagnostics. Furthermore, this polypeptide model system enables the analysis and potentially reliable interpretation of the correlation between chemical structure and immunogenic/antigenic features. PMID- 9346836 TI - Rapid desilylation of oligoribonucleotides at elevated temperatures: cleavage activity in ribozyme-substrate assays. AB - Treatment of 2'-O-silyl-oligoribonucleotides with triethylamine trihydrofluoride in DMF at 55 degrees C for 1 h effected complete desilylation. The product was isolated by a single addition of 1-butanol to the reaction mixture. The resulting RNA was found to be identical with that obtained by traditional desilylation methods as analyzed by HPLC, enzyme digest and ribozyme-substrate assays. PMID- 9346835 TI - Structural effects of glycosylation on the C-terminal pentapeptide of peptide T. AB - Structural effects of glycosylation of the C-terminal pentapeptide fragment of Peptide T (Thr-Thr-Asn-Tyr-Thr) were studied. Because of the inherent flexibility of these molecules, molecular simulations are used to interpret the circular dichroism and nuclear magnetic resonance data acquired for these molecules. N acetyl-glucosamine attached at the Asn residue changes the ensemble average backbone conformation of the peptide and limits the conformational space available to the pentapeptide fragment. Glycosylation changes the type I and III beta-turn propensity of the pentapeptide to a type II turn. Since glycosylation also increases peptide solubility and inhibits peptide degradation in human serum, glycopeptide design may be an efficient approach to stabilize or conformationally modify peptide drug candidates and to create additional diversity in peptide libraries. PMID- 9346838 TI - Enzymatic degradation of various antisense oligonucleotides: monitoring and fragment identification by MECC and ES-MS. AB - Efficacy and sequence specific behaviour of antisense oligonucleotides in biological systems are attenuated by enzymatic degradation, which is predominantly dependent on the oligonucleotide modification. Quantitative data relating to the kinetics and pattern of enzymatic digestion are thus valuable for the interpretation of biological tests with novel antisense oligonucleotides. To study the stability of modified oligonucleotides against nuclease attack, in vitro experiments of enzymatic degradation have been carried out using micellar electrokinetic capillary chromatography (MECC) as a quantitative control and electrospray mass spectrometry (ES-MS) for fragment identification. In contrast to gel electrophoresis, which is commonly applied, monitoring of enzymatic digestion by MECC can be carried out directly from the incubated sample without the need for labeled substrate. Furthermore, exact quantitative analysis becomes possible. Phosphodiester oligonucleotides terminally conjugated with hexaethylene glycol have been prepared to investigate the stability and degradation process of 3'- and 5'-protected oligomers with natural backbones in serum-containing medium. The results demonstrate that 3'-protection is much more effective than 5' protection for nuclease stability, both in fetal calf serum and in human blood serum. To examine the influence of backbone modification on nuclease stability, the digestion of dodecanucleotides containing different numbers of phosphorothioate groups has been investigated by MECC and ES-MS. Degradation rates vary by a factor of approximately 50. Most fragments have been identified and the degradation patterns allow conclusions about the variations of nucleolytic activity with changing substrates. PMID- 9346837 TI - Identification and synthesis of altered peptides modulating T cell recognition of a synthetic peptide antigen. AB - In studies of T cell responses to synthetic peptides we have observed agonist and antagonist activities associated with contaminants identified within the parent synthesis. The synthesis of two candidate analogues implied by a peptide contaminant formed during the synthesis of La 51-58 (IMIKFNRL) has been carried out. The peptide contaminant was 17-18 Da smaller than the parent peptide consistent with a modified asparagine residue at position 6 and so we synthesised both an aspartimide and a nitrile analogue, representing cyclisation or dehydration of the asparagine residue. The candidate aspartimide and nitrile analogues both bound empty MHC class I molecules to form allo determinants recognised by monoclonal antibodies. These results demonstrate that altered synthetic peptides can bind class I MHC molecules and prompt caution in the use of synthetic peptides as a source of immunising antigen. PMID- 9346839 TI - Cleavage of a beta-amyloid precursor sequence by cathepsin D. AB - The identify of the proteases that release beta-amyloid, found in the senile plaques of Alzheimer's disease, from its precursor APP, have not been rigorously identified. As senile plaques contain lysosomal enzymes, and production of some of the amyloidogenic intermediates are inhibited by lysosomotrophic agents, it has been suggested that cathepsins are involved in amyloidogenesis. A synthetic 31-residue peptide overlapping the beta-secretase cleavage site is found to be digested at two mutually exclusive sites, one and three residues on the N terminal side of the N-terminal Asp residue of beta-amyloid. Coupled with the action of aminopeptidases, lysosomal or endosomal cathepsin D could be responsible for generating the N-terminus of beta-amyloid in vivo. PMID- 9346840 TI - Chemical synthesis of polar zipper peptides: motifs for potential protein association in inherited neurodegenerative diseases. AB - Syntheses of peptides containing contiguous repeats of glutamine or asparagine, related to sequences derived from proteins associated with neurodegenerative diseases, are described. Such homo-polymeric peptides are notoriously difficult to prepare due to the formation of aggregated structures during their solid phase synthesis. A novel approach using a randomized array of side chain protected and unprotected residues proved to be an effective strategy for preventing intermolecular peptide-chain association. PMID- 9346841 TI - Transcending the structuralist paradigm in immunology-affinity and biological activity rather than purely structural considerations should guide the design of synthetic peptide epitopes. AB - Synthetic peptides are frequently used to mimic the antigenic sites of proteins. In order to increase the level of mimicry between the peptide and the protein, it is important to understand the structural basis of protein antigenicity. A review of recent crystal structures of antigen-antibody complexes shows that important conformational rearrangements occur in both antigen and antibody during complexation and that many water molecules are located at the complex interface. Both these features are responsible for the low success rate of antigen-antibody docking. The complementarity observed in the complex cannot be predicted from the structure of the free molecules before the occurrence of induced fit and mutual adaptation. The structuralist paradigm assumes that it is possible to understand complex biological recognition phenomena solely in terms of structural data. However, the dynamic component of protein structure requires that both space and time dimensions be included in the description of antigenic specificity. This means that both structural data and activity measurements are required for understanding immunological interactions and for designing synthetic epitopes. The recently developed biosensor technology should greatly facilitate the quantitative measurement of binding interactions and the design of synthetic peptide epitopes. PMID- 9346842 TI - Evaluation of immunodominant epitopes of human T-lymphotropic virus type 1 (HTLV I) using synthetic peptides. AB - Human T-lymphotropic virus type 1 (HTLV-I) causes adult T-cell leukemia/lymphoma (ATLL) and has been associated with a variety of immunologically-mediated diseases. Recently, the immunodominant epitopes of HTLV-I have begun to be defined through the utilization of synthetic peptides and recombinant proteins. Strategies to define the conformational features of immunogenic peptides and design chimeric and multivalent constructs that mimic native viral proteins have provided the opportunity to create an effective synthetic vaccine against HTLV-I infection. An ideal peptide vaccine to be universally immunogenic must incorporate rationally designed antigenic determinants that accurately mimic the corresponding structural architecture found in native proteins and elicit relevant components of the immune system. We have recently designed and tested chimeric and beta-sheet template constructs containing HTLV-I immunodominant peptide motifs that elicit neutralizing antibody responses and overcome genetically restricted immune responses. To further illustrate putative vaccine candidates, HTLV-I env and tax proteins were analyzed using various computer predicted correlates of protein antigenicity, secondary structural predictions, and major histocompatibility complex class I binding motifs. These approaches provide the opportunity to design synthetic peptide vaccines against HTLV-I infection that are based on structurally defined criteria, as well as test the influence of glycosylation on peptide conformation and immunogenicity. PMID- 9346843 TI - Chemoselective approaches to the preparation of peptide dendrimers and branched artificial proteins using unprotected peptides as building blocks. AB - Peptide dendrimers such as Multiple Antigen Peptides (MAPs) are artificial proteins with branched architectures. They hold promise in biochemical and biomedical applications such as synthetic vaccines, serodiagnostics and intracellular delivery of peptides. We have shown that a new design of MAPs containing lipidated built-in adjuvant can be delivered by oral administration to elicit systemic and mucosal immunoglobulins as well as cytotoxic T-lymphocytes. For synthetic vaccines, it is desirable to obtain highly homogeneous preparations. To provide the precision and chemical unambiguity of this class of artificial proteins, we have devised several chemoselective approaches by thiol and carbonyl chemistries to facilitate their synthesis using unprotected peptide segments as building blocks and ligating them to the core matrix. This paper describes the methods of preparation, comparative studies of stability, and presents results of the preparation of antigens containing preformed multiple disulfides. PMID- 9346844 TI - Cyclic peptides as conformationally restricted models of viral antigens: application to foot-and-mouth disease virus. AB - Conformationally restricted cyclic peptide mimics of the antigenic site A of foot and-mouth disease virus serotype C-S8c1 have been designed, first by comparison to the three-dimensional structure of the O1BFS serotype, later more accurately on the basis of X-ray diffraction data from a complex between a linear peptide reproducing site A and an FMDV-derived monoclonal antibody Fab fragment. A variety of cyclization strategies have been attempted, both in solution and in the solid phase, involving disulfide, side chain lactam and head-to-tail arrangements. Preliminary immunological results have shown one of the cyclic disulfide mimics to be a better immunogen than its linear counterpart. PMID- 9346845 TI - Development of an anti-adhesive vaccine for Pseudomonas aeruginosa targeting the C-terminal region of the pilin structural protein. AB - This study describes the development of passive and active vaccines directed at the Pseudomonas aeruginosa pilus adhesin. Passive immunization studies were carried out with P. aeruginosa strain K pilus-specific (PK3B, PK99H) and cross reactive (PAK-13) monoclonal antibodies (MAbs). When A.BY/SnJ mice were passively immunized with a pilus-specific MAb (PK99H), which inhibited pilus-mediated adherence to respiratory epithelial cells, mice challenged with 5 x LD 50 of P. aeruginosa were completely protected while mice were not protected when animals were passively immunized with a pilus specific MAb (PK3B), which did not inhibit pilus adherence to epithilial cells. MAb PAK-13 was found to cross-react with the C-terminal portion of pili of different strains of P. aeruginosa. When mice were passively immunized with MAb PAK-13, subsequent challenge with KB7 (3 x LD50), PAO (8 x LD50) and PAK (3 x LD50) strains of P. aeruginosa resulted in a 70%, 60% and 90% protection of the mice, respectively. MAb PK99H has been previously shown to recognize a linear antigenic epitope consisting of the sequence DEQFIPK. This epitopic peptide was conjugated to protein carriers using different coupling strategies. Use of an appropriate adjuvant and the correct conjugation strategy were critical for raising high affinity antipeptide antisera. In a comparison of Freund's, alum, and Adjuvax, as adjuvants for a peptide-tetanus toxoid conjugate vaccine, highest titers for the synthetic peptide component of the conjugate were obtained with Adjuvax, while highest titers for the carrier protein components were obtained with Freund's. Of the four peptide-conjugates used in this study, only the C-terminal conjugated peptide failed to produce antibodies that bind to native antigen and did not protect mice in active immunization experiments (no survivors at 80 h in the mouse infection model). Conformationally restricted peptide conjugates in which the peptide was conjugated to the carrier at both ends provided better protection in mice challenged with lethal doses of P. aeruginosa than either N- or C-terminal linked peptide-conjugates. The pilus adhesin plays a critical role in P. aeruginosa pathogenesis and this is an excellent vaccine target for either active or passive immunization strategies. PMID- 9346846 TI - The intracellular assembly of antigenic-peptide-class II complexes. AB - The immune system employs remarkable strategies to ensure that foreign antigens, from the most complex pathogens to the simplest proteins, are displayed on the surfaces of cells which are targets of T lymphocyte recognition. At the heart of these strategies is the molecular transformation of a soluble protein antigen to a complex of a small peptide containing the antigenic determinant bound to a cell surface Major Histocompatibility Complex class I or class II protein. This process is termed antigen presentation. Progress in a variety of laboratories over the last several years has yielded a wealth of information about the molecular mechanisms underlying antigen presentation, providing potential new approaches to vaccine design. Here we describe recent studies in our laboratory aimed at elucidating the intracellular site in B lymphocytes in which antigenic peptide-class II complexes are assembled for recognition by helper T cells and the regulation of this assembly process. Our results suggest that processed antigen-class II complexes are assembled in a unique compartment in the endocytic route which contains all the necessary cellular and molecular machinery for assembly and that B cells regulate the assembly process in response to external and internal signals. PMID- 9346847 TI - Prediction of peptide affinity to HLA DR molecules. AB - A method to quantitatively predict peptide binding to HLA DRB1*0401, B1*0101, and B1*1501 has been developed using a dataset of the relative contributions of each of the naturally occurring amino acids in the context of a simplified peptide backbone. The prediction assumed that the relative role of each of the peptide sidechains could be treated independently and could be measured by assaying each of the twenty naturally occurring amino acids at the central eleven positions of a 13 residue peptide previously shown to contain the minimal requirements for high affinity binding to HLA DR proteins. Three separate databases were generated. They were shown to have predictive value when tested on a set of 13 unrelated peptides known to bind the DR proteins with a wide range of apparent affinity. The DRB1*0401 database was tested further by analyzing myelin basic protein. All 13 amino acid peptides containing a hydrophobic amino acid at the third position were synthesized and assayed for binding purified DRB1*0401. In every case, the measured affinity correlated with the predictive values within the experimental error of the assays. Finally, the ability to predict peptide binding to MHC class II molecules was shown to help in identifying T cell determinants. The specificity of DRB1*0401 restricted T cell hybridomas against human serum albumin corresponded to two peptides, predicted, and shown to bind the class II protein with high affinity. PMID- 9346849 TI - Antigenic and immunogenic properties of synthetic peptide-based T-cell determinant polymers. AB - Presentation of T-cell determinants to the immune system in multimeric form has clear advantages and the production of synthetic peptide-based polymers using the solubilisable KS resin described by Goddard et al. [1] provides a method of assembling such polymers and also offers the means for making heteropolymers. The present study investigates the potential of polymeric synthetic peptide constructs in eliciting proliferative T-cell responses to determinants of the influenza virus hemagglutinin. The induction of vigorous CD4+ T-cell immunity was achieved with a polymeric construct containing two different T-cell determinants. The data presented here also highlight the fact that distancing the determinant from the support backbone with appropriate amino acid residues is an important consideration for the success of these polymeric immunogens. This approach may be readily applied in other systems where induction of helper T-cell responses are required. PMID- 9346848 TI - Computer design of T-cell agonist or antagonist glycopeptides: the effect of sugar identity and anomeric configuration on MHC binding. AB - The improved chemical and biological properties of synthetic glycopeptides over peptides suggest their use as T cell agonists or antagonists. Recently, we prepared glycopeptide analogues of major T helper cell epitopic peptides corresponding to rabies virus proteins, and experimentally characterized their ability to bind to MHC class II proteins and stimulate T cell clones to rabies virus. In the current study, we investigated these MHC: peptide interactions by molecular modeling. We obtained structural support for our finding concerning the anomeric specificity of MHC with binding. While alpha-linked glycopeptides can bind to MHC without major alterations in the spatial arrangements and hydrogen bonding pattern of class II-peptide binding, the binding of beta-linked glycopeptides is considerably less favorable due to steric and columbic conflicts. Depending on where the saccharides are positioned along the peptide sequence, the MHC: glycopeptide complex may or may not produce the surface profile required for successful T cell receptor interaction. Application of this approach to other antigenic stimuli offers a good model to "dial in" the necessary sugar identity, length and anomeric configuration, as well as promising amino acid mutation sites, for successful design of T cell agonist or antagonist glycopeptides. PMID- 9346850 TI - Synthetic antigenic peptides as a new strategy for immunotherapy of cancer. AB - Antigens presented by class I of the major histocompatibility complex (MHC) are recognised by the T cell receptor of CD8+ cytolytic effector cells (CTLs), while class II molecules present antigens to CD4+ helper T cells. For both class I and class II molecules, structure and function are linked through the binding of peptides. Consensus or individual sequences have been obtained for naturally processed peptides bound to a variety of class I and class II molecules, revealing the general features of peptides associated with MHC molecules. The interactions between peptides and MHC molecules have been more clearly defined by the characterization of the three dimensional structure of several different MHC molecules. CTLs have been implicated in immune responses against tumors and it is now well documented that some human tumors express specific antigens, which are recognised by CTLs and could potentially be used in immunotherapy protocols. The use of antigenic peptides to elicit a specific and effective CTL response in vivo offers several advantages over the use of other antigenic moieties. Emerging strategies for the safe and effective administration of peptides to humans may lead to their use in the immunological prevention and treatment of cancer. PMID- 9346851 TI - Subunit peptide cancer vaccines targeting activating mutations of the p21 ras proto-oncogene. AB - Activating mutations of the p21 ras proto-oncogene are involved in the development of many common malignancies. Because activating mutations are limited in number, occur within otherwise completely conserved regions, and can be expressed by premalignant lesions, ras is an attractive target for subunit peptide vaccine approaches. Several studies in transplantable tumor models support the possibility that protection against tumors bearing activated ras can be achieved using peptide-based immunogens. We have identified an autochthonous tumor model, A/J mouse lung, which parallels human tumors in the progression of proliferative lesions from premalignant to malignant and which is a very sensitive in vivo system for the detection of activated ras. Although T-cells recognizing a number of activating substitutions can be elicited in this model, peptide immunogens corresponding to the most commonly observed activating mutations are weakly immunogenic. We have engineered a chimeric immunogen incorporating a promiscuous T-cell epitope to enhance the immunogenicity of an oligopeptide corresponding to a weakly immunogenic substitution. These and other challenges associated with developing subunit peptide vaccines to prevent tumors bearing activated ras are discussed. PMID- 9346852 TI - Recombinant epithelial cell mucin (MUC-1) expressed in baculovirus resembles antigenically tumor associated mucin, target for cancer immunotherapy. AB - Epithelial cell mucin encoded by the gene MUC-1, is expressed on several human adenocarcinomas in an aberrantly glycosylated form, and as such it has been identified as the target of human cellular as well as humoral responses. In order to harness this immunity to combat mucin-expressing tumors, various forms of this molecule, synthetic or highly purified, are being tested as possible cancer vaccines. We have expressed MUC-1 in baculovirus, and we report that the recombinant product has important similarities with the MUC-1 expressed on tumors, especially in regard to its aberrant glycosylation. PMID- 9346853 TI - Two antipeptide monoclonal antibodies that recognize adhesive sequences in fibrinogen: identification of antigenic determinants and unrelated sequences using synthetic combinatorial libraries. AB - The fine specificity of two different monoclonal antibodies raised against synthetic peptides, each representing one of the two Arg-Gly-Asp (RGD) sequences in fibrinogen, was examined using synthetic combinatorial libraries (SCLs). The monoclonal antibodies (mAb), mAb LJ-134B/29 and mAb LJ-155B/16, recognize both the immunogenic peptide and native fibrinogen. The specificity of mAb LJ-134B29 was mapped using hexa- and decapeptide positional scanning SCLs (PS-SCLs) and competitive ELISA. The most active amino acids at each position of the two libraries were identified from a single screening. Individual hexa- and decapeptides were synthesized and assayed to determine their binding affinities. The 16 individual hexapeptides represented single and multiple substitutions of the antigenic determinant sequence, -GDSTFE-, eight of which had affinities less than 10nM. Four of the twelve individual decapeptides were found to have binding affinities of approximately 300nM, or nearly three-fold less than the peptide immunogen. A dual-defined hexapeptide library was screened against mAb LJ 155B/16, and individual peptides were obtained through an iterative selection and synthesis process. Surprisingly, one of the most active sequences was Ac-WWYESW NH2 (IC50 = 40nM), which showed no similarity to the sequence of the immunizing peptide. Further mapping of the specificity of this antibody revealed that the antigenic determinant within the peptide immunogen was not completely linear. Recognition of this unrelated sequence by mAb LJ-155B/16 was confirmed in a direct binding assay using biotinylated peptide. The use of SCLs for the elucidation of high affinity peptides recognized by these two antibodies may provide additional information on the molecular mechanisms of fibrinogen binding to different integrin receptors. PMID- 9346854 TI - Idiotype specific peptides bind to the surface immunoglobulins of two murine B cell lymphoma lines, inducing signal transduction. AB - Using a random combinatorial synthetic peptide library method based on a one-bead one-peptide concept for ligand identification (Lam et. al, Nature 1991, 354, 82 84.), idiotype specific peptides were retrieved and optimized for interaction with the cell surface immunoglobulins [IgM(kappa)] of two murine B lymphoma cell lines. Several of the identified peptides were characterized with respect to cell binding and signal transduction. These peptides were able to bind specifically to the surface immunoglobulins of these lymphoma cells. In addition to binding, when synthesized in tetrameric or multimeric forms, the peptides were able to trigger signal transduction resulting in an increase in protein tyrosine phosphorylation. Since D-amino acid peptide libraries were used in some of our efforts to identify binding ligands, several of the idiotype-specific peptides are composed of all D amino acids (e.g. wGeyvmvnG). These findings may have important therapeutic implications for targeted-therapy of B-cell lymphoma as these D-amino acid ligands are more resistant to proteolysis resulting in a prolonged pharmacokinetic disposition in vivo. PMID- 9346855 TI - Heterologous expression, purification, activity and conformational studies of different forms of dianthin 30. AB - Dianthin 30, a ribosome inactivating protein (RIP) from Dianthus caryophyllus, has been expressed in Escherichia coli. Heterologous expression of a deletion mutant dianthin 30 delta 255-270 resulted in the production of a protein identical to carnation mature dianthin 30, including the absence at the carboxy terminal of a putative 16 amino acid long pro-signal peptide. The production of a form of dianthin 30, which includes the pro-signal, is described as well. Both dianthin 30 delta 255-270 and dianthin 30 expressed in E. coli are mainly localized (90%) in the soluble fraction. Dianthin 30 delta 255-270 and dianthin 30 have been purified to homogeneity and were shown to inhibit protein synthesis in vitro with an IC50 of 8 and of 11 ng/ml, respectively. Secondary structure analysis, carried out by circular dichroism spectroscopy, indicated that the naturally occurring and the recombinant forms of dianthin 30 and dianthin 30 delta 255-270 possess the same secondary structure composition, accounting for an alpha + beta type architecture. RIPs as immunotoxins in clinical trial and as mitotoxins in experimental models have been extremely efficacious. In addition, growing evidence indicates their effective use as antiviral agents, including in HIV-1 infection. These data indicate the ability to produce either chemically linked or recombinant fusion proteins with dianthin 30 and cell-binding ligands for production of new reagents for clinical and experimental use. PMID- 9346856 TI - Structure-activity relationships and physico-chemical properties of synthetic lipopeptide inhibitors of PKC. AB - Four synthetic lipopeptides, (K-pm 19,31), (K-pm 19,21,31), (K-pm 19,28,31) and (K-pm 19,21,28,31) with the lysine-palmitoyl (K-pm) residue as a lipophilic moiety, based on the pseudosubstrate sequence 19RFARKGALRQKNV31 (R19-V31), were found to be potent protein kinase C (PKC) inhibitors. However, the lipopeptides (K-pm 19,21,31), (K-pm 19,28,31) and (K-pm 19,21,28,31) were also found to act as protein kinase cAMP-dependent (PKA) inhibitors. Peptide (K-pm 19,31), the least water soluble, is marginally selective towards PKC, unlike the other palmitoyl derivatives studied here. Since the non-palmitoylated analogues (K 19,31), (K-ac 19,31), (K 19,21,31) and (K-ac 19,21,31) were inhibitors of PKC but not of PKA, the palmitoyl moiety must play a role in the specificity of protein kinase inhibition. In vitro, the lipophilic peptides showed greater stability to protease-mediated hydrolysis than the pseudosubstrate peptide depending upon the number of lipophilic (K-pm) residues. CD studies showed that in comparison with the peptide analogues, the remarkable resistance of the pseudosubstrate (R19-V31) to adopt an alpha-helix conformation in TFE, known to be strongly alpha-helix inducing, rules out this structure as the peptide binding conformation to PKC. By contrast, in aqueous media all the peptides show an extended conformation that correlates well with their inhibitory activity. This is in compliance with the crystallographic observation that an extended structure has been observed for the (5-24) PKI peptide inhibitor bound to PKA. PMID- 9346857 TI - The effect of pituitary adenylate cyclase activating polypeptide (PACAP) on amylase secretion from guinea pig pancreatic acini. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a novel hypothalamic peptide, structurally related to vasoactive intestinal peptide (VIP). Our previous study in conscious dogs revealed that PACAP stimulated exocrine pancreatic secretion in a manner different from VIP. The objectives of this study were to characterize the effects of PACAP on amylase secretion and intracellular cAMP production in guinea pig pancreatic acini and to compare them with those of VIP and secretin. PACAP38 and PACAP27 (10(-10)-10(-8)M) stimulated amylase secretion from pancreatic acini in a concentration-related manner. The order of potency for amylase secretion was PACAP27 = VIP > PACAP38. The maximally stimulated amylase secretion by PACAP27 was not enhanced by VIP or secretin, but was synergistically increased by cholecystokinin and A23187. PACAP38 and PACAP27 (10(-2)-10(-7)M) increased intracellular cAMP levels in a concentration-related manner. The potency for cAMP production was PACAP38 = PACAP27 = VIP. These results suggest that PACAP38 and PACAP27, like VIP, directly stimulate amylase secretion from guinea pig pancreatic acini through alterations in cellular cAMP levels. PMID- 9346858 TI - Synthesis, solution structure and biological action of PACAP-related peptide. AB - High quality PACAP-related peptide (PRP), a 29 amino-acid region of the PACAP precursor protein, has been synthesized in quantities sufficient for biological and structural studies. PRP has a distinct biological activity on the gallbladder that is similar to PACAP, but opposite to that of VIP and its related peptide, PHM. Its solution structure has been investigated by circular dichroism spectroscopy and 2D 1H nuclear magnetic resonance spectroscopy. In contrast to the poorly defined structure in aqueous solution alone, the limiting structure, under conditions that mimic a membrane-like environment, possesses stable secondary structure with a helical region between residues 3 and 20, that is terminated by the presence of glycine at residue 21 and is followed by a region of nascent helix. The similarities and differences in the structure of PRP, PACAP27 and GHRH(1-29) are made through comparison of their H alpha chemical shift data and differences in their biological activities assessed. PMID- 9346859 TI - Immunogenicity and conformational properties of an N-linked glycosylated peptide epitope of human T-lymphotropic virus type 1 (HTLV-I). AB - The identification and characterization of epitopes of human T-lymphotropic virus type 1 (HTLV-I), which elicit an effective humoral or cell-mediated immune response, remains a central obstacle to the development of a peptide-based vaccine against the virus infection. The objective of the studies presented here was to examine the influence of N-linked glycosylation on peptide structure and immunogenicity. We engineered the 233-253 sequence of gp46 of HTLV-I to contain an N-acetylglucosamine (GlcNAc) residue at Asn244. Secondary structure prediction using computer algorithms indicated that this peptide may contain a beta-turn at residues 242-246. Recent work with model glycopeptides suggests that beta-turn conformation in peptides may be induced, and probably is stabilized, by the presence of even a single sugar residue. In the present study, the structures of the 233-253 peptide, SC1, and the 233-253(Asn244-GlcNAc) glycopeptide, SC2, were determined. Similar conformation was exhibited by both the glycosylated and nonglycosylated peptide displaying a beta-turn at residues 243-246 and extended chain structure at the peptide/glycopeptide termini. Both peptides were engineered into chimeric constructs with a promiscuous T-cell epitope from measles virus and were used as immunogens in rabbits. Both chimeric peptides were highly immunogenic in rabbits, producing high-titered antibodies as early as primary + three weeks. The antibodies generated against either construct were able to bind to whole virus (ELISA) and to gp46 (radioimmunoprecipitation assay). Additionally, human sera of individuals known to be positive for HTLV-I recognized both the glycosylated and nonglycosylated constructs. It appears that the 233-253 peptide is able to adopt a conformation that mimics the structure in native gp46, and addition of a GlcNAc residue at Asn244 does not affect the conformational preference or stability of this construct; nor does glycosylation alter immunogenicity but instead appears to enhance immune recognition. PMID- 9346861 TI - Conformational studies of the beta-subunit of the high affinity IgE receptor: circular dichroism and molecular modelling. AB - The receptor with high affinity for immunoglobulin E (Fc epsilon RI) on mast cells and basophils plays an important role in mediating many of the pathophysiological phenomena associated with allergy. Fc epsilon RI is a tetrameric complex, alpha beta gamma2, of non-covalently attached subunits: one IgE-binding alpha-subunit with the binding site in the extracellular part of the chain, one beta-subunit and a dimer of disulphide linked gamma-subunits. In the present work, prediction of the three-dimensional structure of the four membrane spanning segments of the beta-subunit has been achieved using rules of helix helix packing arrangements and molecular dynamics calculations. It yielded a four helix bundle with specific Van der Waals interactions between the helices. This four-helix bundle was used as a framework upon which to calculate the conformation of the beta-subunit excluding the C and N terminal cytoplasmic tails, but including the three chains that connect the four helices in the bundle. Separately, these synthetic 11, 17 and 29 residue bridge peptides were examined by circular dichroism (CD) spectroscopy and a degree of alpha-helical content in these bridge peptides was found. Additional molecular modelling of the bridge peptides indicate the central residues of these as the location of the helical moieties. Finally, in the model proposed for the beta-subunit, for each pair of consecutive transmembrane (TM) helices and its bridge peptide, a helix loop-helix-loop-helix motif was found. PMID- 9346860 TI - Solid phase synthesis and immunogenicity of a VP3 peptide from hepatitis A virus. AB - The synthesis of a peptide belonging to the VP3 capsid protein of Hepatitis A virus has been accomplished by the continuous flow Fmoc-polyamide solid phase method. The use of methoxytrimethylbenzenesulphonyl (Mtr) and pentamethylchromansulphonyl (Pmc) as arginine side-chain protecting groups in the presence of tryptophan without lateral protection or protected with t-Boc is discussed. The synthetic VP3 peptide has been administered to mice in different forms: (i) free, (ii) coupled to keyhole limpet hemocyanin, (iii) encapsulated in multilamellar (MLV) liposomes, and (iv) incorporated to a tetrameric branched lysine core. The immune response induced by these preparation is reported. PMID- 9346863 TI - Synthesis and studies on the biophysical activity of human lung surfactant peptide SP-C and its N-terminal fragments. AB - Human lung surfactant peptide SP-C and two of its N-terminal fragments were prepared by SPPS and their biophysical activities investigated in vitro using a pulsating bubble surfactometer. These studies demonstrated that even low doses of the synthetic peptides with the natural human sequence of SP-C in combination with reconstituted lipid mixtures causes a drastic decrease of surface tension. PMID- 9346862 TI - Conformational studies on beta-amyloid protein carboxy-terminal region (residues 34-42): strategic use of amide backbone protection as a structural probe. AB - Analogues of beta-amyloid (32-42) peptide, containing N-(2-hydroxy-4 methoxybenzyl) (Hmb) amide backbone substitutions at various positions have been prepared using fluoren-9-ylmethoxycarbonyl (Fmoc)-polyamide based solid phase peptide synthesis. On-line N alpha-Fmoc deprotection monitoring during assembly exhibited hindered release in the native and beta A(34-42, (Hmb)Gly38) analogue syntheses. No such hindrance was observed during the synthesis of beta A(34-42, (Hmb)Gly37) nor beta A(34-42, (Hmb)Val36). However, the latter contained an exceptionally slow coupling reaction. Cleaved peptides were analysed for solubility in a variety of solvents and insoluble pellets tested for congophilic staining. X-ray analysis of Fmoc (and H-) beta A(34-42) and the corresponding (Hmb)Gly38 analogues as dimethylformamide swollen gels gave very similar structures. Secondary structure prediction and model-building of ordered arrays, compatible with our results, suggest that beta A(34-42) forms a beta-hairpin structure, with the reverse turn at Val36-Gly37-Gly38-Val39 both in solution and on the resin during synthesis. PMID- 9346864 TI - Detection of PCR products from Mycobacterium avium subspecies Paratuberculosis using oligonucleotides containing multiple 2,4-dinitrophenyl reporter groups. AB - A pool of five oligonucleotides has been used to detect the pathogenic organism Mycobacterium avium subspecies paratuberculosis in PCR-amplified DNA from ruminants. The oligonucleotides were labelled at the 5'-end with three dinitrophenyl reporter groups and hybridised to the target DNA, which was fixed to a nylon membrane by ultraviolet irradiation. Colourimetric detection of the PCR product was carried out using an anti-DNP antibody conjugated to horseradish peroxidase or to alkaline phosphatase. Detection with alkaline phosphatase was more sensitive than with horseradish peroxidase but, in both cases, the PCR product could be easily detected. The DNP labelling system offers an economic and effective alternative to biotin, digoxigenin or fluorescein for the detection of PCR-amplified DNA. PMID- 9346865 TI - Equilibrium analysis of the interaction between a synthetic peptide of influenza virus hemagglutinin and monoclonal antibodies. AB - The affinity of interaction between two monoclonal antibodies and a synthetic peptide representing the C-terminal 23 residues of the heavy chain (HA1) of influenza virus hemagglutinin were determined using an air-driven ultracentrifuge. The technique makes use of common laboratory equipment and is based on sound theoretical principles. Because the method does not rely on the solid-phase immobilisation of one of the interacting species, it circumvents problems associated with ELISA-like assays, which, in the case of peptides, may involve the immobilisation of ligand through association of amino acid residues necessary for recognition by antibody. The technique should be applicable to the study of a wide range of ligand-acceptor systems. Because only one of the reagents needs to be pure to allow labelling, prior purification of the biological receptor is not necessary. The method also lends itself to inhibition experiments in which the effects of various homologs on the binding event can be examined in a way which permits an evaluation of potential agonists and antagonists. PMID- 9346866 TI - Rapid and efficient oligonucleotide synthesis with low reagent consumption via a new synthesis column design: preparation of fluorescent dye labelled primers for application in PCR. AB - A low dead-volume, 40 nmole scale column was designed for automated, solid support oligonucleotide synthesis. The LV40 columns are filled with 1000A, high cross link polystyrene beads at the 40 nmole scale. Reducing the unoccupied volume and optimizing the column dimensions allows efficient and fast synthesis on existing commercial synthesizers with low reagent consumption. Three spectrally distinct fluorescent dyes were applied as phosphoramidites in the synthesis of PCR primers. Fluorescent labelled PCR products of the Mfd11 microsatellite locus were analyzed. PMID- 9346867 TI - Spectroscopy and modelling of the cytoplasmic domain of the gamma-subunit of the high affinity immunoglobulin E receptor. AB - The high affinity receptor for IgE, Fc epsilon RI, is responsible for immediate hypersensitivity reactions. In rodents Fc epsilon RI is a tetrameric complex, alpha beta gamma 2 of non-covalently attached subunits: one IgE-binding alpha subunit with the binding site in the extracellular part of the chain, one beta subunit and a dimer of disulphide linked gamma-subunits. Although there is an increasing evidence that the gamma-subunit chains are important signalling proteins that appear to function through a common Tyr-Leu-Tyr-Leu amino acid motif present in their cytoplasmic tails, which link the ligand binding specificity of their associated chains to signal transduction pathways, many questions related to conformation and function of this subunit remain to be answered. In the present work, the 36-residue cytoplasmic domain of the gamma subunit has been synthesized and conformational studies by the combined use of Fourier transform infrared (FTIR), circular dichroism (CD) and nuclear magnetic resonance (NMR) have been performed. Based on the constraints found by these methods, conformational models of the cytoplasmic tail of the gamma-subunit are proposed and discussed. PMID- 9346869 TI - Development of pituitary adenylate cyclase activating polypeptides (PACAPs) specific radioimmunoassay systems and distribution of PACAP-like immunoreactivity in guinea pig tissues. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) specific radioimmunoassay systems have been developed and the distribution of PACAPs in guinea pig tissues has been studied. The antibody against PACAP38 was characterized, using synthetic peptide fragments. It was shown to recognize the C terminal portion with the C-terminal amide group and no cross-reaction was observed with vasoactive intestinal polypeptide (VIP), which has a high homology with PACAP27 and the N-terminal 28 amino acid residues of PACAP38. The antibody against PACAP27 was specific to PACAP27. With the two PACAPs (PACAP38 and PACAP27) specific RIA systems, high concentrations of PACAP38- and PACAP27-like immunoreactivity (LI) were observed in the brain of guinea pigs, especially in the diencephalon (mostly hypothalamus). In all tissues PACAP38-LI was higher than that of PACAP27-LI. Reverse-phase HPLC showed that PACAP38- and PACAP27-LI of tissue extracts were superimposed at the elution position of those of the synthetic peptides, respectively. PMID- 9346868 TI - Chemical synthesis and characterisation of rat chaperonin 10: effect of chain length, ions, heat and N-terminal acetylation on unchaperoned folding into its heptameric form. AB - Recently, the sequence of mitochondrial chaperonin 10 from Rattus norvegicus (rat cpn10), with N-terminal acetylation, has been published. Two syntheses of rat cpn10 were performed, the first using a classical carbodiimide-mediated double coupling protocol (Method A) and the second a more efficient HBTU/HOBT/single coupling procedure (Method B). The latter also involved the application of a capping procedure, using N-(2-chlorobenzyloxycarbonyloxy)succinimide [Z(2-Cl) OSu]. The crude protein from Method A was purified using a two-step isoelectric focusing/RP-HPLC scheme and found to contain a high proportion of a deletion peptide (less Gln60). Conversely, rat cpn10 from Method B was purified to homogeneity by one-step RP-HPLC, using a reversible lipophilic chromatographic probe. The proportion of biologically active heptameric structure was directly related to the purity of the protein and attained 84% with material from Method B. The addition of Ca/Mg ions, pH 7.2, or a heating/cooling cycle increased the proportion of heptamer for less pure protein. Shorter sequences were found not to fold into heptamers, suggesting that aggregation/folding motifs are located in 1 25 and 77-101 regions of rat cpn10. The heptameric cpn10 (Method B) bound correctly to GroEL from E. coli, demonstrating that N-terminal acetylation is not necessary for its folding and binding to bacterial cpn60. PMID- 9346870 TI - Design, synthesis and characterization of bradykinin antagonists via cyclization of the modified backbone. AB - With the aim of synthesizing cyclic antagonists of the nonapeptide hormone bradykinin with minimal side chain modification, we performed backbone to backbone and backbone to side chain cyclization. To probe and compare different strategies for this new kind of cyclization, the branched peptide bonds were formed by both reductive alkylation on the solid phase and by using preformed building units. Lactam bridges between the modified amide groups were formed by the use of the phenylalanine derivatives N(CH2COOH)Phe and N(CH2CH2NH2)Phe. The best results in the formation of the N-alkylamide bond were obtained with the coupling reagent PyBrop. The coupling rate was monitored by estimation of the N terminal Fmoc-group. The cyclization was performed on the solid support. Unexpected difficulties resulted from the instability of the N-alkylamide bond under strong acidic conditions, as used for deprotection and for removal from the resin. We synthesized peptides with backbone to backbone cyclization between positions 2 and 5, as well as backbone to side chain cyclizations between positions 0 and 5, and between 2 and 6. The relatively high biological activities of some of the cyclic analogues support the supposed receptor-bound conformation of bradykinin antagonists with a beta-turn in the N-terminal sequence. PMID- 9346871 TI - NLPR, an agonist of AVP4-8, increases NGF gene expression in memory-impaired rat brain. AB - Oral administration of the tetrapeptide Asn-Leu-Pro-Arg (NLPR) to memory-impaired rats results in improved acquisition and maintenance of behavioural response and also facilitates nerve growth factor (NGF) expression in the brain. It is suggested that NLPR can ameliorate memory disability by promoting NGF gene expression, so implying that NLPR is a potential drug candidate for curing memory impairment. PMID- 9346872 TI - Interindividual differences in 2H8-toluene toxicokinetics assessed by a semiempirical physiologically based model. PMID- 9346875 TI - Transcriptional regulation of mammalian genes in vivo. A tale of two templates. PMID- 9346874 TI - Human genetic affinities for Y-chromosome P49a,f/TaqI haplotypes show strong correspondence with linguistics. AB - Numerous population samples from around the world have been tested for Y chromosome-specific p49a,f/TaqI restriction polymorphisms. Here we review the literature as well as unpublished data on Y-chromosome p49a,f/TaqI haplotypes and provide a new nomenclature unifying the notations used by different laboratories. We use this large data set to study worldwide genetic variability of human populations for this paternally transmitted chromosome segment. We observe, for the Y chromosome, an important level of population genetics structure among human populations (FST = .230, P < .001), mainly due to genetic differences among distinct linguistic groups of populations (FCT = .246, P < .001). A multivariate analysis based on genetic distances between populations shows that human population structure inferred from the Y chromosome corresponds broadly to language families (r = .567, P < .001), in agreement with autosomal and mitochondrial data. Times of divergence of linguistic families, estimated from their internal level of genetic differentiation, are fairly concordant with current archaeological and linguistic hypotheses. Variability of the p49a,f/TaqI polymorphic marker is also significantly correlated with the geographic location of the populations (r = .613, P < .001), reflecting the fact that distinct linguistic groups generally also occupy distinct geographic areas. Comparison of Y-chromosome and mtDNA RFLPs in a restricted set of populations shows a globally high level of congruence, but it also allows identification of unequal maternal and paternal contributions to the gene pool of several populations. PMID- 9346876 TI - A beta-arrestin/green fluorescent protein biosensor for detecting G protein coupled receptor activation. AB - G protein-coupled receptors (GPCR) represent the single most important drug targets for medical therapy, and information from genome sequencing and genomic data bases has substantially accelerated their discovery. The lack of a systematic approach either to identify the function of a new GPCR or to associate it with a cognate ligand has added to the growing number of orphan receptors. In this work we provide a novel approach to this problem using a beta arrestin2/green fluorescent protein conjugate (betaarr2-GFP). It provides a real time and single cell based assay to monitor GPCR activation and GPCR-G protein coupled receptor kinase or GPCR-arrestin interactions. Confocal microscopy demonstrates the translocation of betaarr2-GFP to more than 15 different ligand activated GPCRs. These data clearly support the common hypothesis that the beta arrestin binding of an activated receptor is a convergent step of GPCR signaling, increase by 5-fold the number of GPCRs known to interact with beta-arrestins, demonstrate that the cytosol is the predominant reservoir of biologically active beta-arrestins, and provide the first direct demonstration of the critical importance of G protein-coupled receptor kinase phosphorylation to the biological regulation of beta-arrestin activity and GPCR signal transduction in living cells. The use of betaarr2-GFP as a biosensor to recognize the activation of pharmacologically distinct GPCRs should accelerate the identification of orphan receptors and permit the optical study of their signal transduction biology intractable to ordinary biochemical methods. PMID- 9346877 TI - Increased activity and fidelity of DNA polymerase beta on single-nucleotide gapped DNA. AB - DNA polymerase beta (pol beta) is an error-prone polymerase that plays a central role in mammalian base excision repair. To better characterize the mechanisms governing rat pol beta activity, we examined polymerization on synthetic primer templates of different structure. Steady-state kinetic analyses revealed that the catalytic efficiency of pol beta (kcat/Km,dNTPapp) is strongly influenced by gap size and the presence of a phosphate group at the 5'-margin of the gap. pol beta exhibited the highest catalytic efficiency on 5'-phosphorylated 1-nucleotide gapped DNA. This efficiency was >/=500 times higher than on non-phosphorylated 1 nucleotide and 6-nucleotide (with or without PO4) gapped DNAs and 2,500 times higher than on primer-template with no gaps. The nucleotide insertion fidelity of pol beta, as judged by its ability to form G-N mispairs, was also higher (10-100 times) on 5'-phosphorylated single-nucleotide gapped DNA compared with the other DNA substrates studied. These data suggest that a primary function of mammalian pol beta is to fill 5'-phosphorylated 1-nucleotide gaps. PMID- 9346878 TI - The crystal structure of domain 1 of receptor protein-tyrosine phosphatase mu. AB - Receptor-like protein-tyrosine phosphatases (RPTPs) play important roles in regulating intracellular processes. We have been investigating the regulation and function of RPTPmu, a receptor-like PTP related to the Ig superfamily of cell adhesion molecules. Recently, the crystal structure of a dimer of the membrane proximal domain of RPTPalpha (RPTPalpha D1) was described (Bilwes, A. M., den Hertog, J., Hunter, T., and Noel J. P. (1996) Nature 382, 555-559). Within this crystal structure, the catalytic site of each subunit of the dimer is sterically blocked by the insertion of the N-terminal helix-turn-helix segment of the dyad related monomer. It was proposed that dimerization would lead to inhibition of catalytic activity and may provide a paradigm for the regulation of the RPTP family. We have determined the crystal structure, to 2.3 A resolution, of RPTPmu D1, which shares 46% sequence identity with that of RPTPalpha D1. Although the tertiary structures of RPTPalpha D1 and RPTPmu D1 are very similar, with a root mean square deviation between equivalent Calpha atoms of 1.1 A, the quaternary structures of these two proteins are different. Neither the catalytic site nor the N-terminal helix-turn-helix segment of RPTPmu D1 participates in protein protein interactions. The catalytic site of RPTPmu D1 is unhindered and adopts an open conformation similar to that of the cytosolic PTP, PTP1B (Barford, D., Flint, A. J., and Tonks, N. K. (1994) Science 263, 1397-1404). We propose that dimerization-induced modulation of RPTP activity may not be a general feature of this family of enzymes. PMID- 9346879 TI - The von Hippel-Lindau gene product inhibits vascular permeability factor/vascular endothelial growth factor expression in renal cell carcinoma by blocking protein kinase C pathways. AB - Mutation or loss of function of the von Hippel-Lindau (VHL) tumor suppressor gene is regularly found in sporadic renal cell carcinomas (RCC), well vascularized malignant tumors that characteristically overexpress vascular permeability factor/vascular endothelial growth factor (VPF/VEGF). The wild-type VHL (wt-VHL) gene product acts to suppress VPF/VEGF expression, which is overexpressed when wt VHL is inactive. The present study investigated the pathways by which VHL regulates VPF/VEGF expression. We found that inhibition of protein kinase C (PKC) represses VPF/VEGF expression in RCC cells that regularly overexpress VPF/VEGF. The wt-VHL expressed by stably transfected RCC cells forms cytoplasmic complexes with two specific PKC isoforms, zeta and delta, and prevents their translocation to the cell membrane where they otherwise would engage in signaling steps that lead to VPF/VEGF overexpression. Other experiments implicated mitogen-activated protein kinase (MAPK) phosphorylation as a downstream step in PKC regulation of VPF/VEGF expression. Taken together, these data demonstrate that wt-VHL, by neutralizing PKC isoforms zeta and delta and thereby inhibiting MAPK activation, plays an important role in preventing aberrant VPF/VEGF overexpression and the angiogenesis that results from such overexpression. PMID- 9346881 TI - pH-dependent stability and conformation of the recombinant human prion protein PrP(90-231). AB - A recombinant protein corresponding to the human prion protein domain encompassing residues 90-231 (huPrP(90-231)) was expressed in Escherichia coli in a soluble form and purified to homogeneity. Spectroscopic data indicate that the conformational properties and the folding pathway of huPrP(90-231) are strongly pH-dependent. Acidic pH induces a dramatic increase in the exposure of hydrophobic patches on the surface of the protein. At pH between 7 and 5, the unfolding of hPrP(90-231) in guanidine hydrochloride occurs as a two-state transition. This contrasts with the unfolding curves at lower pH values, which indicate a three-state transition, with the presence of a stable protein folding intermediate. While the secondary structure of the native huPrP(90-231) is largely alpha-helical, the stable intermediate is rich in beta-sheet structure. These findings have important implications for understanding the initial events on the pathway toward the conversion of the normal into the pathological forms of prion protein. PMID- 9346880 TI - Cleavage of single strand RNA adjacent to RNA-DNA duplex regions by Escherichia coli RNase H1. AB - RNase H1 from Escherichia coli cleaves single strand RNA extending 3' from an RNA DNA duplex. Substrates consisting of a 25-mer RNA annealed to complementary DNA ranging in length from 9-17 nucleotides were designed to create overhanging single strand RNA regions extending 5' and 3' from the RNA-DNA duplex. Digestion of single strand RNA was observed exclusively within the 3' overhang region and not the 5' overhang region. RNase H digestion of the 3' overhang region resulted in digestion products with 5'-phosphate and 3'-hydroxyl termini. The number of single strand RNA residues cleaved by RNase H is influenced by the sequence of the single strand RNA immediately adjacent to the RNA-DNA duplex and appears to be a function of the stacking properties of the RNA residues adjacent to the RNA DNA duplex. RNase H digestion of the 3' overhang region was not observed for a substrate that contained a 2'-methoxy antisense strand. The introduction of 3 deoxynucleotides at the 5' terminus of the 2'-methoxy antisense oligonucleotide resulted in cleavage. These results offer additional insights into the binding directionality of RNase H with respect to the heteroduplex substrate. PMID- 9346882 TI - Atypical protein kinase C iota protects human leukemia cells against drug-induced apoptosis. AB - Protein kinase C (PKC) isozymes play distinct roles in cellular function. In human K562 leukemia cells, PKC alpha is important for cellular differentiation and PKC betaII is required for proliferation. In this report, we assess the role of the atypical PKC isoform PKC iota in K562 leukemia cell physiology. K562 cells were stably transfected with expression plasmids containing the cDNA for human PKC iota in sense or antisense orientation to increase or decrease cellular PKC iota levels, respectively. Overexpression or inhibition of expression of PKC iota had no significant effect on the proliferative capacity of K562 cells nor their sensitivity to phorbol myristate acetate-induced cytostasis and megakaryocytic differentiation, suggesting that PKC iota does not play a critical role in these processes. Rather, PKC iota serves to protect K562 cells against drug-induced apoptosis. K562 cells, which are resistant to most apoptotic agents, undergo apoptosis when treated with the protein phosphatase inhibitor okadaic acid (OA). Overexpression of PKC iota leads to increased resistance to OA-induced apoptosis whereas inhibition of PKC iota expression sensitizes cells to OA-induced apoptosis. Overexpression of the related atypical PKC zeta has no protective effect, demonstrating that the effect is isotype-specific. PKC iota also protects K562 cells against taxol-induced apoptosis, indicating that it plays a general protective role against apoptotic stimuli. These data support a role for PKC iota in leukemia cell survival. PMID- 9346883 TI - Protein kinase C phosphorylates the "a" forms of plasma membrane Ca2+ pump isoforms 2 and 3 and prevents binding of calmodulin. AB - Phosphorylation by protein kinase C of the "a" and "b" variants of plasma membrane Ca2+ pump isoforms 2 and 3 was studied. Full-length versions of these isoforms were assembled and expressed in COS cells. Whereas the "a" forms were phosphorylated easily with PKC, isoform 2b was phosphorylated only a little, and isoform 3b was not phosphorylated at all. Phosphorylation of isoforms 2a and 3a did not affect their basal activity, but prevented the stimulation of their activity by calmodulin and their binding to calmodulin-Sepharose. This indicated that phosphorylation prevented activation of these isoforms by preventing calmodulin binding. Based on these results, phosphorylation of the pump with PKC would be expected to increase free intracellular Ca2+ levels in those cells where isoforms 2a and 3a are expressed. PMID- 9346884 TI - Activation of HIV-1 coreceptor (CXCR4) mediates myelosuppression. AB - Chemokines are cytokines that activate and induce the migration of leukocytes. Stroma-derived factor-1 (SDF-1) is a novel chemokine that blocks the entry of T tropic HIV-1 mediated by fusin/CXCR4/LESTR (leukocyte-derived seven-transmembrane domain receptor). In this work we demonstrate that SDF-1 triggers increases in intracellular calcium and inhibits the proliferation of myeloid progenitor cell line 32D. By contrast, SDF-1 neither triggers a calcium response nor affects the proliferation of the myeloid progenitor cell line 32D-GR that is deficient in CXCR4. Responsiveness to SDF-1 was rescued by transfection of 32D-GR cells with a cDNA encoding the human CXCR4. The data indicate that SDF-1 induces myelosuppression by activation of CXCR4. The constitutive production of SDF-1 by bone marrow stromal cells argues for a major role of SDF-1 on the regulation of myelopoiesis. PMID- 9346885 TI - Proinsulin targeting to the regulated pathway is not impaired in carboxypeptidase E-deficient Cpefat/Cpefat mice. AB - Sorting of proinsulin from the trans-Golgi network to secretory granules is critical for its conversion to insulin as well as for regulated insulin secretion. The proinsulin sorting mechanism is unknown. Recently, carboxypeptidase E (CPE) was proposed as a sorting receptor for prohormones. To know whether CPE is implicated in proinsulin sorting, pancreatic islets were isolated from CPE-deficient Cpefat/Cpefat mice and Cpefat/+ controls, pulse labeled ([3H]leucine), and then chased in basal medium (90 min) to examine constitutive secretion followed by medium with secretagogues (60 min) to stimulate regulated secretion. Secretion of labeled proinsulin via the constitutive pathway was <2% even in Cpefat/Cpefat islets. After a 150-min chase, only 13% of radioactivity remained as proinsulin in Cpefat/+ islets compared with 46% in Cpefat/Cpefat islets, reflecting slower conversion. Regulated secretion was stimulated to an equal extent from Cpefat/+ and Cpefat/Cpefat mice with 20% of the total content of labeled (pro)insulin released during the 60-min stimulatory period. It is concluded that in CPE-deficient Cpefat/Cpefat mice, proinsulin is efficiently routed to the regulated pathway and its release can be effectively stimulated by secretagogues. CPE is thus not essential for sorting proinsulin to granules. PMID- 9346886 TI - Evidence that the transfer of hydride ion equivalents between nucleotides by proton-translocating transhydrogenase is direct. AB - The molecular masses of the purified, recombinant nucleotide-binding domains (domains I and III) of transhydrogenase from Rhodospirillum rubrum were determined by electrospray mass spectrometry. The values obtained, 40,273 and 21,469 Da, for domains I and III, respectively, are similar to those estimated from the amino acid sequences of the proteins. Evidently, there are no prosthetic groups or metal centers that can serve as reducible intermediates in hydride transfer between nucleotides bound to these proteins. The transient-state kinetics of hydride transfer catalyzed by mixtures of recombinant domains I and III were studied by stopped-flow spectrophotometry. The data indicate that oxidation of NADPH, bound to domain III, and reduction of acetylpyridine adenine dinucleotide (an NAD+ analogue), bound to domain I, are simultaneous and very fast. The transient-state reaction proceeds as a biphasic burst of hydride transfer before establishment of a steady state, which is limited by slow release of NADP+. Hydride transfer between the nucleotides is evidently direct. This conclusion indicates that the nicotinamide rings of the nucleotides are in close apposition during the hydride transfer reaction, and it imposes firm constraints on the mechanism by which transhydrogenation is linked to proton translocation. PMID- 9346887 TI - Syk and Fyn are required by mouse megakaryocytes for the rise in intracellular calcium induced by a collagen-related peptide. AB - Stimulation of platelets by collagen leads to activation of a tyrosine kinase cascade resulting in secretion and aggregation. We have recently shown that this pathway involves rapid tyrosine phosphorylation of an Fc receptor gamma chain, which contains an immunoreceptor tyrosine-based activation motif (ITAM), enabling interaction with the tandem SH2 domains of the tyrosine kinase Syk. Activation of Syk lies upstream of tyrosine phosphorylation of phospholipase Cgamma2. In the present study we sought to test directly the role of the ITAM/Syk interaction and the role of the Src-related kinases in collagen receptor signaling using mouse megakaryocytes. We demonstrate that the calcium-mobilizing action of a collagen related peptide (CRP) is kinase-dependent, inhibited by the microinjection of the tandem SH2 domains of Syk and abolished in Syk-deficient mice. Furthermore, the CRP response is abolished by the Src family kinase inhibitor PP1 and inhibited in Fyn-deficient mice. In contrast, the calcium response to the G-protein-linked receptor agonist thrombin is not significantly altered under these conditions. These results provide direct evidence of the functional importance of Fyn and Syk in collagen receptor signaling and support the megakaryocyte as a model for the study of proteins involved in this pathway. PMID- 9346888 TI - Cell-specific regulation of expression of plasma-type platelet-activating factor acetylhydrolase in the liver. AB - Platelet-activating factor (PAF) is a potent proinflammatory phospholipid mediator that causes hypotension, increases vascular permeability, and has been implicated in anaphylaxis, septic shock and several other inflammatory responses. PAF is hydrolyzed and inactivated by the enzyme PAF-acetylhydrolase. In the intact rat, a mesenteric vein infusion of lipopolysaccharide (LPS) served as an acute, liver-focused model of endotoxemia. Plasma PAF-acetylhydrolase activity increased 2-fold by 24 h following LPS administration. Ribonuclease protection experiments demonstrated very low levels of plasma-type PAF-acetylhydrolase mRNA transcripts in the livers of saline-infused rats; however, 24 h following LPS exposure, a 20-fold induction of PAF-acetylhydrolase mRNA was detected. In cells isolated from endotoxin-exposed rat livers, Northern blot analyses demonstrated that Kupffer cells but not hepatocytes or endothelial cells were responsible for the increased PAF-acetylhydrolase mRNA levels. In Kupffer cells, plasma-type PAF acetylhydrolase mRNA was induced by 12 h, peaked at 24 h, and remained substantially elevated at 48 h. Induction of neutropenia prior to LPS administration had no effect on the increase in PAF-acetylhydrolase mRNA seen at 24 h. Although freshly isolated Kupffer cells contain barely detectable levels of plasma-type PAF-acetylhydrolase mRNA, when Kupffer cells were established in culture, PAF-acetylhydrolase expression became constitutively activated concomitant with cell adherence to the culture plates. Alterations in plasma-type PAF-acetylhydrolase expression may constitute an important mechanism for elevating plasma PAF-acetylhydrolase levels and an important component in minimizing PAF-mediated pathophysiology in livers exposed to endotoxemia. PMID- 9346889 TI - A regulatory element within a coding exon modulates keratin 18 gene expression in transgenic mice. AB - Multiple tissue-specific, DNase-hypersensitive sites are correlated with known or potential regulatory regions of the human keratin 18 (K18) gene. One of these sites is found within exon 6, close to a potential AP-1 binding site. Footprint analysis confirmed that this site is capable of binding c-Jun and c-Fos in vitro. However, exon 6 can stimulate expression of a reporter gene driven by the K18 proximal promoter independent of AP-1 in F9 cells and additionally modulates AP-1 responsiveness when in combination with an intron enhancer. Analysis in transgenic mice and by transient transfections of mutant forms of the K18 gene showed that exon 6 contributes to the expression of the K18 gene. However, substitution of part of exon 6 with the corresponding part of the keratin 19 gene which lacks an AP-1 site decreased but did not destroy the regulatory activity of the exon. Furthermore, this mutation did not alter either the tissue specificity or the position-independent and copy number-dependent behavior of the K18 gene. In contrast, a frameshift mutation within exon 6 dramatically decreased the expression of the gene. K18 RNA expression from the frameshift mutation was less than 10% of the wild type K18 transgene. This decline in expression was the result of a combination of decreased stability of mutant K18 RNA and the creation of a negative regulatory element that can interact with the first intron regulatory elements and actively suppress K18 expression. These results demonstrate that a protein-coding portion of the K18 gene also has a regulatory function. PMID- 9346890 TI - Purification and cloning of hepatocyte growth factor activator inhibitor type 2, a Kunitz-type serine protease inhibitor. AB - Hepatocyte growth factor (HGF) activator is a serine protease responsible for proteolytic activation of HGF in response to tissue injury and thus plays an important role in the regulation of biological functions of HGF in regenerating tissue. We previously purified an inhibitor of HGF activator (HGF activator inhibitor type 1, HAI-1) from the conditioned medium of a human stomach carcinoma cell line MKN45 and cloned its cDNA. HAI-1 is a novel member of the Kunitz family of serine protease inhibitors. In the present study, we purified a second type of HGF activator inhibitor (HAI-2) from the conditioned medium of MKN45 cells and molecularly cloned its cDNA. The cDNA sequence revealed that HAI-2 is derived from a precursor protein of 252 amino acids and contains two Kunitz domains, indicating that HAI-2 is also a member of the Kunitz family of serine protease inhibitors. The primary translation product of HAI-2 has a hydrophobic sequence in the COOH-terminal region, suggesting that, like HAI-1, HAI-2 is produced in a membrane-associated form and secreted in a proteolytically truncated form. Because HAI-2 and HAI-1 are potent inhibitors specific for HGF activator, they may be involved in regulation of proteolytic activation of HGF in injured tissues. PMID- 9346892 TI - A mutant truncated protein disulfide isomerase with no chaperone activity. AB - A mutant human protein disulfide isomerase with the COOH-terminal 51 amino acid residues deleted (abb'a') has been expressed in Escherichia coli. Its secondary structures are very similar to those of the native bovine enzyme. The mutant enzyme shows neither peptide binding ability nor chaperone activity in assisting the refolding of denatured D-glyceraldehyde-3-phosphate dehydrogenase but keeps most of the catalytic activities for reduction of insulin and isomerization of scrambled ribonuclease. It assists the reactivation of denatured and reduced proteins containing disulfide bonds, acid phospholipase A2, and lysozyme to different levels, which are significantly lower than those by the native bovine enzyme. PMID- 9346891 TI - A single precursor protein for ferrochelatase-I from Arabidopsis is imported in vitro into both chloroplasts and mitochondria. AB - Ferrochelatase is the last enzyme of heme biosynthesis and in higher plants is found in both chloroplasts and mitochondria. We have isolated cDNAs for two isoforms of ferrochelatase from Arabidopsis thaliana, both of which are imported into isolated chloroplasts. In this paper we show that ferrochelatase-I is also imported into isolated pea mitochondria with approximately the same efficiency as into chloroplasts. Processing of the precursor was observed with both chloroplast stroma and mitochondrial matrix extracts. This was inhibited by EDTA, indicating it was due to the specific processing proteases. The specificity of import was verified by the fact that the mitochondrial preparation did not import the precursor of the light-harvesting chlorophyll a/b protein precursor or the precursor of porphobilinogen deaminase, an earlier enzyme of tetrapyrrole biosynthesis, both of which are exclusively chloroplast-located. Furthermore, import of ferrochelatase-I precursor into mitochondria was inhibited by valinomycin, but this had no effect on its import into chloroplasts. Thus a single precursor molecule is recognized by the import machinery of the two organelles. The implications for the targeting of ferrochelatase in a possible protective role against photooxidative stress are discussed. PMID- 9346893 TI - Complex subunit assembly of neuronal voltage-gated K+ channels. Basis for high affinity toxin interactions and pharmacology. AB - Neurons require specific patterns of K+ channel subunit expression as well as the precise coassembly of channel subunits into heterotetrameric structures for proper integration and transmission of electrical signals. In vivo subunit coassembly was investigated by studying the pharmacological profile, distribution, and subunit composition of voltage-gated Shaker family K+ (Kv1) channels in rat cerebellum that are labeled by 125I-margatoxin (125I-MgTX; Kd, 0.08 pM). High-resolution receptor autoradiography showed spatial receptor expression mainly in basket cell terminals (52% of all cerebellar sites) and the molecular layer (39% of sites). Sequence-directed antibodies indicated overlapping expression of Kv1. 1 and Kv1.2 in basket cell terminals, whereas the molecular layer expressed Kv1.1, Kv1.2, Kv1.3, and Kv1.6 proteins. Immunoprecipitation experiments revealed that all 125I-MgTX receptors contain at least one Kv1.2 subunit and that 83% of these receptors are heterotetramers of Kv1.1 and Kv1.2 subunits. Moreover, 33% of these Kv1.1/Kv1.2-containing receptors possess either an additional Kv1.3 or Kv1.6 subunit. Only a minority of the 125I MgTX receptors (<20%) seem to be homotetrameric Kv1.2 channels. Heterologous coexpression of Kv1.1 and Kv1.2 subunits in COS-1 cells leads to the formation of a complex that combines the pharmacological profile of both parent subunits, reconstituting the native MgTX receptor phenotype. Subunit assembly provides the structural basis for toxin binding pharmacology and can lead to the association of as many as three distinct channel subunits to form functional K+ channels in vivo. PMID- 9346894 TI - FKBP12 binds the inositol 1,4,5-trisphosphate receptor at leucine-proline (1400 1401) and anchors calcineurin to this FK506-like domain. AB - The immunophilin FKBP12 is one of the most abundant and conserved proteins in biology. It is the primary receptor for the immunosuppressant actions of the drug FK506 in whose presence FKBP12 binds to and inhibits calcineurin, disrupting interleukin formation in lymphocytes. The physiologic functions of FKBP12 are less clear, although the protein has been demonstrated to physiologically interact with the inositol 1,4,5-trisphosphate receptor (IP3R), the ryanodine receptor, and the type 1 transforming growth factor beta receptor. We now report that FKBP12 binds the IP3R at residues 1400-1401, a leucyl-prolyl dipeptide epitope that structurally resembles FK506. We further demonstrate that binding to IP3R at this site enables FKBP12 to interact with calcineurin, presumably to anchor the phosphatase to IP3R and modulate the receptor's phosphorylation status. We propose that FK506 promotes an FKBP12-calcineurin interaction by mimicking structurally similar dipeptide epitopes present within proteins that use FKBP12 to anchor calcineurin to the appropriate physiologic substrates. PMID- 9346895 TI - Glycogenin-2, a novel self-glucosylating protein involved in liver glycogen biosynthesis. AB - Glycogenin is a self-glucosylating protein involved in the initiation phase of glycogen biosynthesis. A single mammalian gene had been reported to account for glycogen biogenesis in liver and muscle, the two major repositories of glycogen. We describe the characterization of novel forms of glycogenin, designated glycogenin-2 (GN-2), encoded by a second gene that is expressed preferentially in certain tissues, including liver, heart, and pancreas. Cloning of cDNAs encoding glycogenin-2 indicated the existence of multiple species, including three liver forms (GN-2alpha, GN-2beta, and GN-2gamma) generated in part by alternative splicing. Overall, GN-2 has 40-45% identity to muscle glycogenin but is 72% identical over a 200-residue segment thought to contain the catalytic domain. GN 2 expressed in Escherichia coli or COS cells is active in self-glucosylation assays, and self-glucosylated GN-2 can be elongated by skeletal muscle glycogen synthase. Antibodies raised against GN-2 produced in E. coli recognized proteins of Mr approximately 66,000 present in extracts of rat liver and in cultured H4IIEC3 hepatoma cells. In H4IIEC3 cells, most of the GN-2 was present as a free protein but some was covalently associated with glycogen fractions and was only released by treatment with alpha-amylase. H4IIEC3 cells also expressed the muscle form of glycogenin (glycogenin-1), which was attached to a chromatographically separable glycogen fraction. PMID- 9346896 TI - Polarized apical targeting directed by the signal/anchor region of simian virus 5 hemagglutinin-neuraminidase. AB - To examine the possibility of independent cytoplasmic/transmembrane domain-based apical sorting, we have investigated paramyxovirus SV5 hemagglutinin neuraminidase (HN), a type II membrane protein with a small N-terminal signal/anchor region. In SV5-infected Madin-Darby canine kidney (MDCK) cells, >90% of HN is found on the apical surface. We have expressed chimeric proteins in which the N terminus of HN, including its signal/anchor region, is attached to a (normally cytosolic) reporter pyruvate kinase (PK). PK itself expressed immediately downstream from a cleavable signal peptide was converted to a 58-kDa N-linked glycosylated form, which was secreted predominantly (80%) to the basolateral surface of MDCK cells. By contrast, stably expressed PK chimeras, now anchored as type II membrane proteins with either the first 48 or 72 amino acids of HN, received similar N-linked glycosylation, yet exhibited polarized transport with a preferentially (75%) apical distribution. These results suggest that the N terminal signal/anchor region of HN contains independent sorting information for apical specific targeting in MDCK cells. PMID- 9346897 TI - Mu and delta opioid receptors are differentially desensitized by the coexpression of beta-adrenergic receptor kinase 2 and beta-arrestin 2 in xenopus oocytes. AB - The Xenopus oocyte expression system was used to test the hypothesis that homologous opioid receptor desensitization results from receptor phosphorylation by G protein-coupled receptor kinases. Activation of delta (DOR), mu (MOR) opioid, or beta2-adrenergic receptors increased K+ conductance in oocytes coexpressing the G protein-gated inwardly rectifying K+ channel subunits GIRK1 and GIRK4, and the intrinsic rate of desensitization was small. Coexpression of beta-adrenergic receptor kinase 2 (beta-ARK2) and beta-arrestin 2 (beta-arr2) synergistically produced a rapid desensitization of both DOR and beta2-adrenergic receptor signaling with a t1/2 < 4 min. beta-ARK2 and beta-arr2 more slowly desensitized MOR responses; a similar synergistic effect on MOR required 2-3 h of agonist treatment. DOR mutants lacking serine and threonine residues at the end of the cytoplasmic tail coupled effectively to GIRK channels but were insensitive to beta-ARK2 and beta-arr2. However, a DOR mutant having serine residues mutated to alanine in the third cytoplasmic loop was indistinguishable in coupling and desensitization from the wild type DOR. These studies establish that opioid receptors can be regulated by beta-ARK2 and beta-arr2 and that a portion of the COOH terminus of DOR enhances sensitivity to this modulation. PMID- 9346899 TI - Antibodies capable of releasing diphtheria toxin in response to the low pH found in endosomes. AB - Diphtheria toxin (DT) undergoes a rapid conformational change in response to the acidity encountered within endosomes. That transition is integral to the passage of its catalytic domain into the cytosol and thus its lethal action. The importance of this translocation mechanism led us to develop several monoclonal antibodies that bind DT at neutral pH but spontaneously release the toxin when critical epitopes denature or unfold upon lowering the pH to 4.5-5.5. Hybridomas were selected using a microtiter plate assay that measured the pH-dependent detachment of antibody from immobilized toxin. The acid-sensitive epitopes involved were on the catalytic, transmembrane, and receptor binding domains of DT. This pH-induced disruption of the binding of toxin to these monoclonal antibodies was analyzed by sedimentation velocity ultracentrifugation. Antibody combining sites were fully occupied at pH 5.5, partially bound at pH 5.0, and totally empty at pH 4.5. It was estimated that the Ka for antibody-toxin binding was approximately 1000-fold lower at pH 5.0 than at neutral pH. This novel acid triggered release mechanism provides a basis for delivery of antibody-bound toxin into cells accompanied by its immediate dissociation as the complex enters acidic vesicles. PMID- 9346898 TI - Cold-sensitive mutants G680V and G691C of Dictyostelium myosin II confer dramatically different biochemical defects. AB - Cold-sensitive myosin mutants represent powerful tools for dissecting discrete deficiencies in myosin function. Biochemical characterization of two such mutants, G680V and G691C, has allowed us to identify separate facets of myosin motor function perturbed by each alteration. Compared with wild type, the G680V myosin exhibits a substantially enhanced affinity for several nucleotides, decreased ATPase activity, and overoccupancy or creation of a novel strongly actin-binding state. The properties of the novel strong binding state are consistent with a partial arrest or pausing at the onset of the mechanical stroke. The G691C mutant, on the other hand, exhibits an elevated basal ATPase indicative of premature phosphate release. By releasing phosphate without a requirement for actin binding, the G691C can bypass the part of the cycle involving the mechanical stroke. The two mutants, despite having alterations in glycine residues separated by only 11 residues, have dramatically different consequences on the mechanochemical cycle. PMID- 9346900 TI - Intracellular targeting with low pH-triggered bispecific antibodies. AB - Bispecific antibodies were designed to deliver a reversibly bound ligand into target cells and then spontaneously release it upon passage into acidified vesicles. These reagents were assembled by coupling monoclonal antibodies that recognize acid-sensitive epitopes on diphtheria toxin to cell type-specific monoclonal antibodies. The dual binding capacity of the bispecific antibodies was confirmed by delivery of 125I-diphtheria toxin to target molecules present on intact cells. Bispecific antibodies directed against transferrin receptors on human cells were loaded with toxin and tested for cytotoxicity. The mutant diphtheria toxins CRM107 and CRM45 were used since their inability to bind cell receptors renders them ordinarily nontoxic. Their full cytotoxic potential, however, was restored via bispecific antibody-mediated delivery and release within low pH intracellular vesicles. Cytotoxicity was shown to be specific by blocking receptor sites and to be acidification-dependent by protection using NH4Cl to raise endosomal pH. Kinetics for inhibition of cellular protein synthesis was identical for native diphtheria toxin and the bispecific antibody. CRM107 combination. The rate of inhibition (t1/2 = 20 min) indicated that release of CRM107 from the antibody combining site was fast, and its toxic action was unimpeded by this delivery mechanism. PMID- 9346901 TI - TRAM-1, A novel 160-kDa thyroid hormone receptor activator molecule, exhibits distinct properties from steroid receptor coactivator-1. AB - Nuclear hormone receptors (NRs) are ligand-dependent transcription factors that regulate target gene transcription. We report the molecular cloning and characterization of a novel human cDNA encoding TRAM-1, a thyroid hormone receptor activator molecule, a approximately 160-kDa protein homologous with SRC 1/TIF2, by far-Western-based expression screening. TRAM-1 binds to thyroid hormone receptor (TR) and other NRs in a ligand-dependent manner and enhances ligand-induced transcriptional activity of TR. The AF-2 region in NRs has been thought to play a critical role in mediating ligand-dependent transactivation by the interaction with coactivators. Surprisingly, TRAM-1 retains strong ligand dependent interaction with an AF-2 mutant of TR (E457A), while SRC-1 fails to interact with this mutant. Furthermore, we identified a critical TRAM-1 binding site in rat TRbeta1 outside of AF-2, as TRAM-1 shows weak ligand-dependent interaction with a helix 3 ligand binding domain TR mutant (K288A), compared with SRC-1. These results suggest that TRAM-1 is a coactivator that may exhibit its activity by interacting with subdomains of NRs other than the AF-2 region, in contrast to SRC-1/TIF2. PMID- 9346902 TI - Structural determinants in AUF1 required for high affinity binding to A + U-rich elements. AB - AUF1 is an RNA-binding protein that contains two nonidentical RNA recognition motifs (RRMs). AUF1 binds to A + U-rich elements (AREs) with high affinity. The binding of AUF1 to AREs is believed to serve as a signal to an mRNA-processing pathway that degrades mRNAs encoding many cytokines, oncoproteins, and G protein coupled receptors. Because the ARE binding activity of AUF1 appears central to the regulation of many important genes, we analyzed the domains of the protein that are important for this activity. Examination of the RNA binding affinity of various AUF1 mutants suggests that both RRMs may be required for binding to the human c-fos ARE. However, the two RRMs together are not sufficient. Highest affinity binding of AUF1 to an ARE requires an alanine-rich region of the N terminus and a short glutamine-rich region in the C terminus. In addition, the N terminus is required for dimerization of AUF1. However, AUF1 binds an ARE as a hexameric protein. Thus, protein-protein interactions are important for high affinity ARE binding activity of AUF1. PMID- 9346903 TI - The enzymatic and non-enzymatic roles of protein-disulfide isomerase in apolipoprotein B secretion. AB - Secretion of apolipoprotein B (apoB) from mammalian cells requires the presence of functional microsomal triglyceride transfer protein (MTP). We previously reported that co-expressing the human intestinal form of apoB, B48, with both subunits of human MTP in oleate-treated Sf21 cells led to a dramatic induction of B48 secretion. Deletion mutagenesis studies showed that the cysteine-enriched amino terminus of apoB was necessary for the MTP responsiveness (Gretch, D. G., Sturley, S. L., Wang, L., Dunning, A., Grunwald, K. A. A., Wetterau, J. R., Yao, Z., Talmud, P., and Attie, A. D. (1996) J. Biol. Chem. 271, 8682-8691). We therefore hypothesized that the small subunit of MTP, protein-disulfide isomerase (PDI), plays a role in apoB secretion by facilitating correct disulfide bond formation. To determine whether the enzymatic activities of PDI are important for MTP-stimulated apoB secretion, the wild type PDI subunit was replaced with an active site mutant, mPDI (Cys36 --> Ser/Cys380 --> Ser), lacking both disulfide shuffling and redox activities. MTP containing mPDI was fully functional in promoting apoB and triglyceride secretion. Therefore, the shufflase and redox activities of PDI are not necessary for the function of MTP. Since PDI exists in large molar excess over the other subunit of MTP, the role of free PDI (independent of the MTP complex) was investigated. PDI or mPDI was co-expressed with B48 and B17, a fragment encompassing the amino-terminal 17% of apoB. Mutant PDI significantly and specifically reduced the accumulation of the B17 and B48 both intracellularly and in the culture medium. The reduction was partially eliminated by the protease inhibitor N-acetyl-leucyl-leucyl-norleucinal, consistent with rapid co- or post-translational degradation of apoB in the presence of mPDI. Treating the cells with oleate reversed the effect of mPDI on B48 secretion in a dose-dependent manner, but had no effect on B17. IN CONCLUSION: 1) the role of PDI in the MTP complex involves functions other than its known enzymatic activities; 2) one or both of the enzymatic activities of free PDI is/are important for the MTP-independent steps of apoB secretion; 3) oleate can affect apoB secretion at high physiological concentrations and compensate for the insufficiency of PDI activities. PMID- 9346904 TI - Inactivation of dehydratase [4Fe-4S] clusters and disruption of iron homeostasis upon cell exposure to peroxynitrite. AB - Phagocytes produce both nitric oxide and superoxide as components of the oxidative defense against pathogens. Neither molecule is likely at physiological concentrations to kill cells. However, two of their reaction products, hydrogen peroxide and peroxynitrite, are strong oxidants, cell-permeant, and toxic. Hydrogen peroxide generates oxidative DNA damage, while the primary mechanism of toxicity of peroxynitrite has not yet been determined. Recent in vitro studies indicated that peroxynitrite is capable of oxidizing the [4Fe-4S] clusters of a family of dehydratases (Hausladen, A., and Fridovich, I. (1994) J. Biol. Chem. 269, 29405-29408; Castro, L., Rodriguez, M., and Radi, R. (1994) J. Biol. Chem. 269, 29409-29415). We demonstrate here that peroxynitrite at 1% of its lethal dose almost fully inactivated the labile dehydratases in Escherichia coli. The rate at which peroxynitrite inactivated the clusters substantially exceeded the rate at which it oxidized thiols or spontaneously decomposed. These results suggest that these dehydratases may be primary targets of peroxynitrite in vivo. Another consequence of the cluster damage was the release of 100 microM iron into the cytosol. During phagocytosis, this intracellular free iron could increase lethal DNA damage by hydrogen peroxide or protein modification by additional peroxynitrite. In response to peroxynitrite challenges, E. coli rapidly sequestered the intracellular free iron using an undefined scavenging system. The iron-sulfur clusters were more gradually repaired by a process that drew iron from its iron-storage proteins. These are likely to be critical events in the struggle between phagocyte and pathogen. PMID- 9346906 TI - Conditional inhibition of the mitogen-activated protein kinase cascade by wortmannin. Dependence on signal strength. AB - Phosphoinositide (PI) 3-kinase and the mitogen-activated protein (MAP) kinase cascades are activated by many of the same ligands. Several groups have reported involvement of PI 3-kinase in the activation of Erk1 and Erk2, whereas many other groups have shown that activation of Erk1 and Erk2 is not sensitive to inhibitors of PI 3-kinase such as wortmannin. Here we show that wortmannin inhibition of the MAP kinase pathway is cell type- and ligand-specific. Wortmannin blocks platelet derived growth factor (PDGF)-dependent activation of Raf-1 and the MAP kinase cascade in Chinese hamster ovary cells, which have few PDGF receptors, but has no significant effect on Erk activation in Swiss 3T3 cells, which have high levels of PDGF receptors. However, wortmannin blocks activation of Erk proteins if Swiss 3T3 cells are stimulated with lower, physiological levels of PDGF. These results suggest that PI 3-kinase is in an efficient pathway for activation of MAP kinase, but that MAP kinase can be stimulated by a redundant pathway when a large number of receptors are activated. We present evidence that a protein kinase C family member downstream of phospholipase Cgamma is involved in the redundant pathway. PMID- 9346907 TI - STT4 is an essential phosphatidylinositol 4-kinase that is a target of wortmannin in Saccharomyces cerevisiae. AB - Wortmannin is a natural product that inhibits signal transduction. One target of wortmannin in mammalian cells is the 110-kDa catalytic subunit of phosphatidylinositol 3-kinase (PI 3-kinase). We show that wortmannin is toxic to the yeast Saccharomyces cerevisiae and present genetic and biochemical evidence that a phosphatidylinositol 4-kinase (PI 4-kinase), STT4, is a target of wortmannin in yeast. In a strain background in which stt4 mutants are rescued by osmotic support with sorbitol, the toxic effects of wortmannin are similarly prevented by sorbitol. In contrast, in a different strain background, STT4 is essential under all conditions and wortmannin toxicity is not mitigated by sorbitol. Overexpression of STT4 confers wortmannin resistance, but overexpression of PIK1, a related PI 4-kinase, does not. In vitro, the PI 4 kinase activity of STT4, but not of PIK1, was potently inhibited by wortmannin. Overexpression of the phosphatidylinositol 4-phosphate 5-kinase homolog MSS4 conferred wortmannin resistance, as did deletion of phospholipase C-1. These observations support a model for a phosphatidylinositol metabolic cascade involving STT4, MSS4, and phospholipase C-1 and provide evidence that an essential product of this pathway is the lipid phosphatidylinositol 4,5 bisphosphate. PMID- 9346905 TI - In vivo regulation of CrkII and CrkL proto-oncogenes in the uterus by insulin like growth factor-I. Differential effects on tyrosine phosphorylation and association with paxillin. AB - Changes in CrkII and CrkL phosphorylation are associated with insulin-like growth factor receptor activation in cultured cells. We examined whether similar changes also occur following administration of recombinant human insulin-like growth factor-I to the intact animal. In female rats starved overnight, CrkL phosphorylation was significantly increased 12 min after insulin-like growth factor-I administration. Tyrosine phosphorylation of CrkII was not detectable in either control or treated animals. Paxillin, a 65-70-kDa phosphoprotein containing high affinity binding sites common for the Src homology 2 (SH2) domains of CrkII and CrkL, was observed in both CrkII and CrkL immunoprecipitates. Insulin-like growth factor-I treatment stimulated the association of CrkII with paxillin. In contrast, the same treatment resulted in the dissociation of the CrkL-paxillin complex. Similar effects of insulin-like growth factor-I treatment on the association of CrkL with tyrosine phosphorylated paxillin were observed in fibroblasts overexpressing CrkL. This study demonstrates that the activation of the insulin-like growth factor-I receptor induces changes in the tyrosine phosphorylation and protein-protein interactions of the Crk proteins in vivo. The different responses of CrkL and CrkII to insulin like growth factor-I receptor activation suggest distinct roles for these two adapter proteins in signal transduction. PMID- 9346908 TI - TbetaRI phosphorylation of Smad2 on Ser465 and Ser467 is required for Smad2-Smad4 complex formation and signaling. AB - Mothers against Dpp-related or Smad proteins are essential components of serine/threonine kinase receptor signaling pathways that are regulated by phosphorylation. Recently, it was demonstrated that Smad2 interacts transiently with and is a direct substrate of the transforming growth factor-beta (TGF-beta) type I receptor, TbetaRI. Phosphorylation sites on Smad2 were localized to a carboxyl-terminal fragment containing three serine residues at positions 464, 465, and 467. In this report, we show that TbetaRI specifically phosphorylates Smad2 on serines 465 and 467. Serine 464 is not a site of phosphorylation, but is important for efficient phosphorylation of Smad2. Phosphorylation at both sites is required to mediate association of Smad2 with Smad4 in mammalian cells, while in yeast, Smad2 interacts directly with Smad4 and does not require phosphorylation. Mutation of either serine residue 465 or 467 prevents dissociation of Smad2 from activated TbetaRI and blocks TGF-beta-dependent signaling and Smad2 transcriptional activity. These results indicate that receptor-dependent phosphorylation of Smad2 on serines 465 and 467 is required in mammalian cells to permit association with Smad4 and to propagate TGF-beta signals. PMID- 9346909 TI - Nine L-type amino acid residues confer full 1,4-dihydropyridine sensitivity to the neuronal calcium channel alpha1A subunit. Role of L-type Met1188. AB - Pharmacological modulation by 1,4-dihydropyridines is a central feature of L-type calcium channels. Recently, eight L-type amino acid residues in transmembrane segments IIIS5, IIIS6, and IVS6 of the calcium channel alpha1 subunit were identified to substantially contribute to 1,4-dihydropyridine sensitivity. To determine whether these eight L-type residues (Thr1066, Gln1070, Ile1180, Ile1183, Tyr1490, Met1491, Ile1497, and Ile1498; alpha1C-a numbering) are sufficient to form a high affinity 1,4-dihydropyridine binding site in a non-L type calcium channel, we transferred them to the 1, 4-dihydropyridine-insensitive alpha1A subunit using site-directed mutagenesis. 1,4-Dihydropyridine agonist and antagonist modulation of barium inward currents mediated by the mutant alpha1A subunits, coexpressed with alpha2delta and beta1a subunits in Xenopus laevis oocytes, was investigated with the two-microelectrode voltage clamp technique. The resulting mutant alpha1A-DHPi displayed low sensitivity for 1,4 dihydropyridines. Analysis of the 1,4-dihydropyridine binding region of an ancestral L-type alpha1 subunit previously cloned from Musca domestica body wall muscle led to the identification of Met1188 (alpha1C-a numbering) as an additional critical constituent of the L-type 1,4-dihydropyridine binding domain. The introduction of this residue into alpha1A-DHPi restored full sensitivity for 1,4-dihydropyridines. It also transferred functional properties considered hallmarks of 1, 4-dihydropyridine agonist and antagonist effects (i.e. stereoselectivity, voltage dependence of drug modulation, and agonist-induced shift in the voltage-dependence of activation). Our gain-of-function mutants provide an excellent model for future studies of the structure-activity relationship of 1, 4-dihydropyridines to obtain critical structural information for the development of drugs for neuronal, non-L-type calcium channels. PMID- 9346910 TI - Dynamics of [Ca2+] in the endoplasmic reticulum and cytoplasm of intact HeLa cells. A comparative study. AB - We have measured the [Ca2+] in the endoplasmic reticulum ([Ca2+]er) of intact HeLa cells at both 22 degrees C and 37 degrees C using endoplamsic reticulum targeted, low Ca2+ affinity aequorin reconstituted with coelenterazine n. Aequorin consumption was much slower at 22 degrees C, and this allowed performing a much longer study of the dynamics of [Ca2+]er. The steady-state [Ca2+]er (500 600 microM) was not modified by the temperature, although both the rates of pumping and leak were decreased at 22 degrees C. The behavior of both [Ca2+]er and cytoplasmic [Ca2+] ([Ca2+]c) after the addition of increasing concentrations of agonists and/or Ca2+-ATPase inhibitors, or following incubation in Ca2+-free medium were compared. We show that agonists induce a fast but relatively small decrease in [Ca2+]er, which is enough to produce a sharp increase in [Ca2+]c. Termination of Ca2+ release is controlled by feedback inhibition of the inositol 1,4,5-trisphosphate receptors by [Ca2+]c, a mechanism that appears to be designed to release the minimum amount of Ca2+ necessary to produced the required [Ca2+]c signal. We also show that Ca2+ release is inhibited progressively when [Ca2+]er decreases below a threshold of about 150 microM, even in the absence of Ca2+ pumping or -Ca2+-c increase. This effect is consistent with a regulation of the inositol 1,4,5-trisphosphate-gated channels by [Ca2+]er. PMID- 9346911 TI - Partial characterization of the auxiliary factors involved in apolipoprotein B mRNA editing through APOBEC-1 affinity chromatography. AB - APOBEC-1-catalyzed apolipoprotein B (apoB) mRNA editing requires auxiliary factors, but the number and functions of these factors are unknown. We have partially purified the editing activity from extracts of a McArdle cell line overexpressing His6-hemagglutinin-tagged, rat APOBEC-1 using metal-chelating affinity chromatography. The 1,200-fold purification achieved by this approach was partially dependent on exogenously added RNA containing a mooring sequence for editosome assembly. Affinity-purified editing activity could be separated by 300 mM NaCl extraction into two fractions, a salt-resistant fraction (editing fraction 1; EF1) and a salt-soluble fraction (EF2). Neither EF1 nor EF2 alone could edit apoB RNA, but when added together they reconstituted full editing activity. Previously identified candidate auxiliary factors including the p66/p44 apoB RNA binding proteins and the presumptive editosome assembly factor p240 were all present in the affinity-purified editing complex. Moreover, virtually all of p66, p240, and APOBEC-1 were present in EF1, whereas p44 was quantitatively recovered in EF2. This is the first demonstration that p66 and p44 can bind to apoB RNA independently of one another. In addition, 100- and 55-kDa apoB RNA cross-linking proteins have been identified in the APOBEC-1 affinity-purified material. RNA competition studies demonstrated that p100, p66, and p55 bound selectively to apoB RNA, whereas p44 had general RNA cross-linking characteristics. The data underscore the multiplicity of auxiliary factors potentially involved in apoB RNA editing and suggest an editosome far more complicated than may have been previously appreciated. PMID- 9346912 TI - Local folding of the N-terminal domain of Escherichia coli RecA controls protein protein interaction. AB - To obtain structural information about the self-association of the protein RecA, we studied urea denaturation of RecA by circular dichroism spectroscopy and gel filtration. Gel filtration analysis showed that urea at low concentrations, 1.0 1.2 M, dissociated the RecA oligomer to almost a monomeric state prior to the unfolding of each molecule. Upon treatment with 1.0 M urea, the circular dichroism spectrum showed a decrease in the alpha-helical content of RecA. A similar decrease was observed in the absence of urea for RecA at an extremely low protein concentration; the RecA oligomer dissociated to an almost completely monomeric state. The properties of RecA at low urea concentrations were similar to those of a truncated RecA lacking the first 33 N-terminal residues (Delta33RecA). Addition of a synthetic peptide corresponding to the 33 N-terminal residues to Delta33RecA increased the alpha-helical content. These results suggest that local folding of the N-terminal domain is coupled to protein-protein interactions of monomeric RecA, which are involved in the regulation of filament formation. The dissociation constant for interaction between RecA monomers was determined from the ellipticity data to be 0.1 microM. PMID- 9346913 TI - Heterologous pleckstrin homology domains do not couple IRS-1 to the insulin receptor. AB - Pleckstrin homology (PH) domains occur in many signaling proteins, including substrates for the insulin receptor tyrosine kinase (IRS proteins). Based on the hypothesis that PH domains may have a common function such as membrane targeting we tested the ability of PH domains from other signaling molecules to link IRS-1 to the insulin receptor. Chimeric IRS-1 proteins containing a homologous PH domain derived from other IRS proteins (IRS-2 or Gab-1) were tyrosine phosphorylated normally in response to insulin. In contrast, heterologous PH domains from the beta-adrenergic receptor kinase, phospholipase Cgamma, or spectrin failed to mediate tyrosine phosphorylation of chimeric IRS-1 proteins, even in cells expressing high levels of insulin receptor. Moreover, IRS-1 proteins containing heterologous PH domains did not bind phosphorylated NPEY motifs derived from the insulin receptor, suggesting that the presence of these structures interfered with the function of the adjacent PTB binding domain. Thus, tyrosine phosphorylation of IRS-1 by the insulin receptor specifically requires a PH domain derived from IRS proteins. PMID- 9346914 TI - The interaction of the molecular chaperone, alpha-crystallin, with molten globule states of bovine alpha-lactalbumin. AB - Small heat shock proteins function in a chaperone-like manner to prevent the precipitation of proteins under conditions of stress (e. g. heat). alpha Crystallin, the major mammalian lens protein, is a small heat shock protein. The mechanism of chaperone action of these proteins is poorly understood. In this paper, the conformational state of a protein when it forms a high molecular weight complex with alpha-crystallin is investigated by examining, using NMR spectroscopy and size exclusion high performance liquid chromatography, the interaction of alpha-crystallin with alpha-lactalbumin and its various intermediately folded (molten globule) states. The complex is formed following reduction of alpha-lactalbumin by dithiothreitol in the presence of alpha crystallin, and this interaction has been monitored in real time by 1H NMR spectroscopy. It is concluded that alpha-crystallin interacts with a disordered molten globule state of alpha-lactalbumin while it is on an irreversible pathway toward aggregation and precipitation. alpha-Crystallin does not interact, however, with molten globule states of alpha-lactalbumin that are stable in solution, e.g. the reduced and carboxyamidated species. It is proposed that alpha crystallin distinguishes between the various molten globule states of alpha lactalbumin on the basis of the lifetimes of these states, i.e. the protein must be in a disordered molten globule state for a significant length of time and on the pathway to aggregation and precipitation for interaction to occur. PMID- 9346915 TI - Activation of acid sphingomyelinase by interleukin-1 (IL-1) requires the IL-1 receptor accessory protein. AB - The cytokine interleukin-1 (IL-1) plays an important role in inflammation and regulation of immune responses, but the mechanisms of its signal transduction and cell activation processes are incompletely understood. Ceramide generated by sphingomyelinases (SMases) is known to function as an important second messenger molecule in the signaling pathway of IL-1 and tumor necrosis factor. To investigate the activation of SMases by IL-1, we used an IL-1 receptor type I (IL 1RI)-positive EL4 thymoma cell line, which is defective in IL-1R accessory protein (IL-1RAcP) expression. In this cell line (EL4D6/76), tumor necrosis factor induced ligand/receptor internalization, NFkappaB nuclear translocation, IL-2 production, and the activation of neutral (N)-SMase and acid (A)-SMase. In contrast, stimulation with IL-1 resulted only in the activation of N-SMase whereas ligand/receptor internalization, NFkappaB translocation, IL-2 production, and activation of A-SMase were not detected. Transfection of this functionally defective EL4D6/76 with IL-1RAcP cDNA restored these functions. These data suggest that A-SMase activity is strongly linked with the internalization of IL 1RI mediated by IL-1RAcP and that A-SMase and N-SMase are activated by different pathways. PMID- 9346917 TI - Purification and characterization of HisP, the ATP-binding subunit of a traffic ATPase (ABC transporter), the histidine permease of Salmonella typhimurium. Solubility, dimerization, and ATPase activity. AB - The nucleotide-binding subunit, HisP, of the histidine permease, a traffic ATPase (ABC transporter), has been purified as a soluble protein and characterized. Addition of a 6-histidine extension (HisP(His6)) allows a rapid and effective metal affinity purification, giving a 30-fold purification with a yield of 50%. HisP(his6) is indistinguishable from underivatized HisP when incorporated into the permease membrane-bound complex, HisQMP2. Purified HisP(his6) has a strong tendency to precipitate; 5 mM ATP and 20% glycerol maintain it in solution at a high protein concentration. HisP(his6) is active as a dimer, binds ATP with a Kd value of 205 microM, and hydrolyzes it at a rate comparable to that of HisQMP2; in contrast to the latter, it does not display cooperativity for ATP. HisP(his6) has been characterized with respect to substrate and inhibitor specificity and various physico-chemical characteristics. Its pH optimum is 7 and it requires a cation for activity, with Co2+ and Mn2+ being more effective than Mg2+ at lower concentrations but inhibitory in the higher concentration range. In contrast to the intact complex, HisP(his6) is not inhibited by vanadate but is inhibited by N ethylmaleimide. Neither the soluble receptor, HisJ, nor the transport substrate, histidine, has any effect on the activity. PMID- 9346918 TI - Direct evidence for an important role of sphingomyelinase in ultraviolet-induced activation of c-Jun N-terminal kinase. AB - Sphingomyelinase (SMase) and its product ceramide have recently attracted a great deal of attention because of their possible role in the signal transduction pathway. However, the role of sphingomyelinase in UV-induced c-June N-terminal kinase (JNK) activation is still unclear. Thus, we investigated this issue directly using a genetic SMase-deficient (2 approximately 3% residual acid SMase activity) lymphoblast cell line, MS1418. The results showed that while UV irradiation markedly induces JNK activation in a normal human lymphoblast cell line, JY, it induces only weak JNK activation in MS1418 cells. This difference of JNK response to UV irradiation between these two cell lines was further observed in time course and dose-response studies. In contrast, 12-O-tetradecanoylphorbol 13-acetate-induced JNK activation could be observed in both JY and MS1418 cells. Furthermore, significant JNK activation can be observed in MS1418 cells by exposure of the cells to SMase or C2-ceramide, whereas phospholipase A2 or phospholipase C did not show significant induction of JNK activity, and C2 dihydroceramide and sphingosine induce only much weaker JNK activation in MS1418 cells than that by C2-ceramide. These data demonstrated that SMase plays an essential role in UV-induced JNK activation. PMID- 9346916 TI - A novel assay reveals a role for soluble N-ethylmaleimide-sensitive fusion attachment protein in mannose 6-phosphate receptor transport from endosomes to the trans Golgi network. AB - Soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein (alpha SNAP) is a soluble protein that enables the NSF ATPase to associate with membranes and facilitate membrane trafficking events. Although NSF and alpha-SNAP have been shown to be required for many membrane transport processes, their role in the transport of mannose 6-phosphate receptors from endosomes to the trans Golgi network was not established. We present here a novel in vitro assay that monitors the transport of cation-dependent mannose 6-phosphate receptors between endosomes and the trans Golgi network. The assay relies on the trans Golgi network localization of tyrosine sulfotransferase and monitors transport of mannose 6-phosphate receptors engineered to contain a consensus sequence for modification by this enzyme. Using this new assay we show that alpha-SNAP strongly stimulates transport in reactions containing limiting amounts of cytosol. Together with alpha-SNAP, NSF can increase the extent of transport. These data show that alpha-SNAP, a soluble component of the SNAP receptor machinery, facilitates transport from endosomes to the trans Golgi network. PMID- 9346919 TI - Short term effects of leptin on hepatic gluconeogenesis and in vivo insulin action. AB - Long term administration of leptin decreases caloric intake and fat mass and improves glucose tolerance. Here we examine whether leptin acutely regulates peripheral and hepatic insulin action. Recombinant mouse leptin (0.3 mg/kg.h, Leptin +) or vehicle (Leptin -) were administered for 6 h to 4-month-old rats (n = 20), and insulin (3 milliunits/kg.min) clamp studies were performed. During physiologic hyperinsulinemia (plasma insulin approximately 65 microunits/ml), the rates of whole body glucose uptake, glycolysis, and glycogen synthesis and the rates of 2-deoxyglucose uptake in individual tissues were similar in Leptin - and Leptin +. Post-absorptive hepatic glucose production (HGP) was similar in the two groups. However, leptin enhanced insulin's inhibition of HGP (4.1 +/- 0.7 and 6.2 +/- 0.7 mg/kg.min; p < 0.05). The decreased HGP in the Leptin + group was due to a marked suppression of hepatic glycogenolysis (0.7 +/- 0.1 versus 4.1 +/- 0.6 mg/kg.min, in Leptin + versus Leptin -, respectively; p < 0.001), whereas the % contribution of gluconeogenesis to HGP was markedly increased (82 +/- 3% versus 36 +/- 4% in Leptin + and Leptin -, respectively; p < 0.001). At the end of the 6 h leptin infusion, the hepatic abundance of glucokinase mRNA was decreased, whereas that of phosphoenolpyruvate carboxykinase mRNA was increased compared with Leptin -. We conclude that an acute increase in plasma leptin 1) enhances insulin's ability to inhibit HGP, 2) does not affect peripheral insulin action, and 3) induces a redistribution of intrahepatic glucose fluxes and changes in the gene expression of hepatic enzymes that closely resemble those of fasting. PMID- 9346920 TI - The endoplasmic reticulum can act as a functional Ca2+ store in all subcellular regions of the pancreatic acinar cell. AB - Stimulation of pancreatic acinar cells raises [Ca2+]i via Ca2+ release from inositol-1,4,5-trisphosphate (InsP3)-sensitive intracellular Ca2+ stores, generally considered to reside within the endoplasmic reticulum (ER). However, with physiological doses of cholinergic agonists, the [Ca2+]i increase is localized to the apical (secretory) pole of the cell, leading to suggestions that zymogen (secretory) granules themselves may constitute an InsP3-sensitive Ca2+ store responsible for localized Ca2+ release. We have therefore re-investigated whether the ER in pancreatic acinar cells is capable of acting as a functional Ca2+ store in all, or only some, cellular regions. In streptolysin O permeabilized cells, the ER accumulated up to 25 mmol of 45Ca2+ per liter ER volume by an ATP-dependent, thapsigargin-sensitive, process. This tracer Ca2+ uptake was dependent on ambient (loading) [Ca2+], as was the intra-ER free [Ca2+], assessed by imaging the fluorescence of Magfura-2 within the Ca2+ stores. Comparison of free and total intra-ER [Ca2+] indicated that 200-300 Ca2+ ions are bound within the ER lumen for every Ca2+ ion remaining free. Subcellular analysis showed that ER stores in all regions of the permeabilized cell took up Ca2+ at loading [Ca2+] between 60 nM and 1 microM. Thapsigargin released Ca2+ from stores in all cellular regions, as did InsP3. Immunofluorescence with antibodies against sarco(endo)plasmic reticulum-2b type Ca2+,Mg2+-ATPase or calreticulin confirmed that ER Ca2+ stores were present throughout the cytoplasm. In summary, these results clearly show that the endoplasmic reticulum can act as a functional Ca2+ store in all regions of the acinar cell, including the apical pole. PMID- 9346921 TI - Activation of p38 mitogen-activated protein kinase by signaling through G protein coupled receptors. Involvement of Gbetagamma and Galphaq/11 subunits. AB - Various extracellular stimuli activate three classes of mitogen-activated protein kinases (MAPKs): extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 MAPK. In mammalian cells, p38 MAPK is activated by endotoxins, inflammatory cytokines, and environmental stresses. We show here that p38 MAPK is also activated upon stimulation of G protein-coupled receptors (Gq/G11-coupled m1 and Gi-coupled m2 muscarinic acetylcholine and Gs-coupled beta-adrenergic receptors) in human embryonal kidney 293 cells. The activation of p38 MAPK through the m2 and beta-adrenergic receptors was completely inhibited by coexpression of Galphao, whereas the activation by the m1 receptor was only partially inhibited. Furthermore, we show that overexpression of Gbetagamma or a constitutively activated mutant of Galpha11, but not Galphas and Galphai, can stimulate p38 MAPK. These results suggest that the signal from the m2 and beta adrenergic receptors to p38 MAPK is mediated by Gbetagamma, whereas the signal from the m1 receptor is mediated by both Gbetagamma and Galphaq/11. PMID- 9346922 TI - The CCAAT box binding factor, NF-Y, is required for thyroid hormone regulation of rat liver S14 gene transcription. AB - Triiodothyronine (T3) activates rat liver S14 gene transcription through T3 receptors (TRbeta) binding distal thyroid hormone response elements located between -2.8 and -2.5 kilobase pairs upstream from the transcription start site. Previous studies suggested that proximal promoter elements located between -220 to -80 base pairs upstream from the 5' end of the S14 gene were involved in hormone activation of the S14 gene. This report identifies an inverted CCAAT box (or Y box) at -104ATTGG-100 as a core cis-regulatory element. Gel shift studies using rat liver nuclear proteins show that at least three CCAAT-binding factors interact with this region as follows: NF-Y and c/EBP-related proteins formed major complexes, whereas NF-1/CTF forms a minor complex in gel shift assay. Mutation of the Y box indicated that loss of NF-Y binding, but not c/EBP or NF-1, correlated closely with a decline in basal activity and a loss of T3-mediated transactivation. Substitution of the S14 Y box in reporter genes with elements binding only NF-Y elevated basal activity and T3-mediated transactivation, whereas substitution with elements binding c/EBP-related proteins or SP1 displayed low basal activity and T3-mediated transactivation. These studies indicate that NF-Y and TRbeta functionally interact to confer T3 control to the S14 gene. PMID- 9346923 TI - The catalytic mechanism of mammalian adenylyl cyclase. Equilibrium binding and kinetic analysis of P-site inhibition. AB - The mechanism of P-site inhibition of adenylyl cyclase has been probed by equilibrium binding measurements using 2'-[3H]deoxyadenosine, a P-site inhibitor, and by kinetic analysis of both the forward and reverse reactions (i.e. cyclic AMP and ATP synthesis, respectively). There is one binding site for 2' deoxyadenosine per C1/C2 heterodimer; the Kd is 40 +/- 3 microM. Binding is observed only in the presence of one of the products of the adenylyl cyclase reaction, pyrophosphate (PPi). A substrate analog, Ap(CH2)pp (alpha,beta methylene adenosine 5'-triphosphate), and cyclic AMP compete for the P-site in the presence of PPi, but P-site analogs do not compete for substrate binding (in the absence of PPi). Kinetic analysis indicates that release of products from the enzyme is random. These facts permit formulation of a model for the adenylyl cyclase reaction, for which we provide substantial kinetic support. We propose that P-site analogs act as dead-end inhibitors of product release, stabilizing an enzyme-product (E-PPi) complex by binding at the active site. Although product release is random, cyclic AMP dissociates from the enzyme preferentially. Release of PPi is slow and partially rate-limiting. PMID- 9346925 TI - SH2/SH3 adaptor proteins can link tyrosine kinases to a Ste20-related protein kinase, HPK1. AB - Ste20-related protein kinases have been implicated as regulating a range of cellular responses, including stress-activated protein kinase pathways and the control of cytoskeletal architecture. An important issue involves the identities of the upstream signals and regulators that might control the biological functions of mammalian Ste20-related protein kinases. HPK1 is a protein serine/threonine kinase that possesses a Ste20-like kinase domain, and in transfected cells activates a protein kinase pathway leading to the stress activated protein kinase SAPK/JNK. Here we have investigated candidate upstream regulators that might interact with HPK1. HPK1 possesses an N-terminal catalytic domain and an extended C-terminal tail with four proline-rich motifs. The SH3 domains of Grb2 bound in vitro to specific proline-rich motifs in the HPK1 tail and functioned synergistically to direct the stable binding of Grb2 to HPK1 in transfected Cos1 cells. Epidermal growth factor (EGF) stimulation did not affect the binding of Grb2 to HPK1 but induced recruitment of the Grb2.HPK1 complex to the autophosphorylated EGF receptor and to the Shc docking protein. Several activated receptor and cytoplasmic tyrosine kinases, including the EGF receptor, stimulated the tyrosine phosphorylation of the HPK1 serine/threonine kinase. These results suggest that HPK1, a mammalian Ste20-related protein serine/threonine kinase, can potentially associate with protein-tyrosine kinases through interactions mediated by SH2/SH3 adaptors such as Grb2. Such interaction may provide a possible mechanism for cross-talk between distinct biochemical pathways following the activation of tyrosine kinases. PMID- 9346924 TI - Destabilization of peptide binding and interdomain communication by an E543K mutation in the bovine 70-kDa heat shock cognate protein, a molecular chaperone. AB - We have compared 70-kDa heat shock cognate protein (Hsc70) isolated from bovine brain with recombinant wild type protein and mutant E543K protein (previously studied as wild type in our laboratory). Wild type bovine and recombinant protein differ by posttranslational modification of lysine 561 but interact similarly with a short peptide (fluorescein-labeled FYQLALT) and with denatured staphylococcal nuclease-(Delta135-149). Mutation E543K results in 4. 5-fold faster release of peptide and lower stability of complexes with staphylococcal nuclease-(Delta135-149). ATP hydrolysis rates of the wild type proteins are enhanced 6-10-fold by the addition of peptide. The E543K mutant has a peptide stimulated hydrolytic rate similar to that of wild type protein but a higher unstimulated rate, yielding a mere 2-fold enhancement. All three versions of Hsc70 possess similar ATP-dependent conformational shifts, and all show potassium ion dependence. These data support the following model: (i) in the presence of K+, Mg2+, and ATP, the peptide binding domain inhibits the ATPase; (ii) binding of peptide relieves this inhibition; and (iii) the E543K mutation significantly attenuates the inhibition by the peptide binding domain and destabilizes Hsc70 peptide complexes. PMID- 9346926 TI - Glutathione peroxidase protects against peroxynitrite-mediated oxidations. A new function for selenoproteins as peroxynitrite reductase. AB - There is a requirement for cellular defense against excessive peroxynitrite generation to protect against DNA strand breaks and mutations and against interference with protein tyrosine-based signaling and other protein functions due to formation of 3-nitrotyrosine. Here, we demonstrate a role of selenium containing enzymes catalyzing peroxynitrite reduction using glutathione peroxidase (GPx) as an example. GPx protected against the oxidation of dihydrorhodamine 123 by peroxynitrite more effectively than ebselen (2-phenyl-1,2 benzisoselenazol-3(2H)-one), a selenoorganic compound exhibiting a high second order rate constant for the reaction with peroxynitrite, 2 x 10(6) M-1 s-1. Carboxymethylation of selenocysteine in GPx by iodoacetate led to the loss of "classical" glutathione peroxidase activity but maintained protection against peroxynitrite-mediated oxidation. The maintenance of protection by GPx against peroxynitrite requires GSH as reductant. When peroxynitrite was infused to maintain a 0.2 microM steady-state concentration, GPx in the presence of GSH, but neither GPx nor GSH alone, effectively inhibited the hydroxylation of benzoate by peroxynitrite. Under these steady-state conditions peroxynitrite did not cause the loss of classical GPx activity. GPx, like selenomethionine, protected against protein 3-nitrotyrosine formation in human fibroblast lysates, shown in Western blots. The formation of nitrite rather than nitrate from peroxynitrite was enhanced by GPx or by selenomethionine. The results demonstrate a novel function of GPx and potentially of other selenoproteins containing selenocysteine or selenomethionine, in the GSH-dependent maintenance of a defense line against peroxynitrite-mediated oxidations, as a peroxynitrite reductase. PMID- 9346927 TI - rRNA maturation as a "quality" control step in ribosomal subunit assembly in Dictyostelium discoideum. AB - In Dictyostelium discoideum, newly assembled ribosomal subunits enter polyribosomes while they still contain immature rRNA. rRNA maturation requires the engagement of the subunits in protein synthesis and leads to stabilization of their structure. Maturation of pre-17 S rRNA occurs only after the newly formed 40 S ribosomal particle has entered an 80 S ribosome and participated at least in the formation of one peptide bond or in one translocation event; maturation of pre-26 S rRNA requires the presence on the 80 S particle of a peptidyl-tRNA containing at least 6 amino acids. Newly assembled particles that cannot fulfill these requirements for structural reasons are disassembled into free immature rRNA and ribosomal proteins. PMID- 9346928 TI - Characterization of a putative helix-loop-helix motif in nucleotide excision repair endonuclease, XPG. AB - Complementation group G of xeroderma pigmentosum (XPG) is one of the most rare and pathophysiologically heterogeneous forms of this inherited disease. XPG patients exhibit varying phenotypes, from having a very mild defect in DNA repair to being severely affected, and a few cases are also associated with the neurological degeneracy and growth retardation of Cockayne's syndrome. The XPG gene encodes a 134-kDa nuclear protein that is essential for the incision steps of nucleotide excision repair. XPG protein contains a putative helix-loop-helix (HLH) motif in the region that is most conserved among the members of structure specific endonuclease family. To establish the functional significance of the HLH motif, we used several approaches, including theoretical modeling, functional complementation assay, structure-specific endonuclease assay, and DNA binding assay. A secondary structure of the motif was predicted by energy minimization and the Monte Carlo simulation and empirically proven using the circular dichroism to contain a high content of alpha-helix. When an XPG mutant lacking the HLH was overexpressed in UV135 cells, which have defects in the hamster homolog of XPG, the mutant gene failed to confer to the hamster cells the resistance to UV light. A recombinant XPG protein lacking the HLH motif was purified from insect cells and tested for a structure-specific endonuclease activity. The mutant protein failed to cleave the flap strand. A recombinant peptide containing the HLH (amino acids 758-871) was expressed in and purified from bacteria, tested for DNA binding activity, and found to bind to a DNA substrate with the flap structure. These results suggest that the HLH motif is required for the catalytic and DNA binding activities of XPG. PMID- 9346931 TI - Two independent nuclear localization signals are present in the DNA-binding high mobility group domains of SRY and SOX9. AB - SRY and SOX9, members of the family of high-mobility group (HMG) domain transcription factors, are both essential for testis formation during human embryonic development. The HMG domain is a DNA-binding and DNA-bending motif comprising about 80 amino acid residues. It has been shown that SRY and SOX9 are nuclear proteins. Using normal or mutant SRY-beta-galactosidase and SOX9-beta galactosidase fusion proteins in transfection studies involving COS-7 cells, we have identified two nuclear localization signals (NLSs) within the HMG domains of both proteins that can independently direct the fusion proteins into the nucleus. Only mutational inactivation of both NLS motifs resulted in complete exclusion of the fusion proteins from the nucleus. The NLS sequences are located at the N and C termini of the HMG domain and are a bipartite NLS motif and a basic cluster NLS motif, respectively. Both NLS motifs are conserved in the HMG domains of other transcription factors. The implications of the present results are discussed regarding (a) the apparent dual function of certain basic amino acid residues in the HMG domain of SRY in both DNA binding and in nuclear localization and (b) the possible control of SOX9 in early gonadal differentiation at the level of nuclear translocation. PMID- 9346930 TI - The main protein of the aggregation factor responsible for species-specific cell adhesion in the marine sponge Microciona prolifera is highly polymorphic. AB - Species-specific cell recognition in sponges, the oldest living metazoans, is based on a proteoglycan-like aggregation factor. We have screened individual sponge cDNA libraries, identifying multiple related forms for the aggregation factor core protein (MAFp3). Northern blots show the presence in several human tissues of transcripts strongly binding a MAFp3-specific probe. The open reading frame for MAFp3 is not interrupted in the 5' direction, revealing variable protein sequences that contain numerous introns equally spaced. We have studied tissue histocompatibility within a sponge population, finding 100% correlation between rejection behavior and the individual-specific restriction fragment length polymorphism pattern using aggregation factor-related probes. PCR amplifications with specific primers showed that at least some of the MAFp3 forms are allelic and distribute in the population used. A pronounced polymorphism is also observed when analyzing purified aggregation factor in polyacrylamide gels. Protease digestion of the polymorphic glycosaminoglycan-containing bands indicates that glycans are also responsible for the variability. The data presented reveal a high polymorphism of aggregation factor components, which matches the elevated sponge alloincompatibility, suggesting an involvement of the cell adhesion system in sponge allogeneic reactions. PMID- 9346932 TI - Cloning and characterization of lung-endothelial cell adhesion molecule-1 suggest it is an endothelial chloride channel. AB - Lung-endothelial cell adhesion molecule-1 (Lu-ECAM-1) is an endothelial cell surface molecule that mediates adhesion of metastatic melanoma cells to lung endothelium. Here we analyze the organization of the Lu-ECAM-1 protein complex, report the sequence of Lu-ECAM-1 cDNAs, and reveal a novel function of the protein. Lu-ECAM-1 immunopurified from bovine aortic endothelial cells (BAEC) consists of tightly associated glycoproteins of 90, 38, and 32 kDa, with minor components of 130 and 120 kDa. We present evidence that all of these protein species are encoded by a single open reading frame whose initial translation product is proteolytically processed to yield the other products. Correct processing in vitro was demonstrated by transfection of the longest cDNA into human embryonic kidney 293 cells; immunoblot analysis showed that the approximately 120-kDa precursor gave rise to 90- and 38-kDa products. RNA blots of BAEC mRNA detected messages in agreement with the sizes of the cDNA clones in addition to several of high molecular weight. DNA blot analysis showed that Lu ECAM-1 is conserved throughout its length in all mammals tested, usually as a single or low copy gene. In the bovine, Lu-ECAM-1 protein is 88% identical to a calcium-dependent chloride channel described recently in tracheal epithelium, Ca CC. Probes for Lu-ECAM-1 mRNA and protein confirmed the presence of a homolog in this tissue. We show that messages for both proteins are present in lung while only Ca-CC is present in trachea and only Lu-ECAM-1 is present in BAEC. These results suggest that endothelial cells express a chloride channel that is related to, but distinct from, that expressed in tracheal epithelium. They further suggest that an adhesion molecule can also be a chloride channel. PMID- 9346933 TI - Primary structure, developmental expression, and immunolocalization of the murine laminin alpha4 chain. AB - The complete primary structure of the mouse laminin alpha4 chain was derived from cDNA clones. The translation product contains a 24-residue signal peptide preceding the mature alpha4 chain of 1,792 residues. Northern analysis on whole mouse embryos revealed that the expression was weak at day 7, but it later increased and peaked at day 15. In adult tissues the strongest expression was observed in lung and cardiac and skeletal muscles. Weak expression was also seen in other adult tissues such as brain, spleen, liver, kidney, and testis. By in situ hybridization of fetal and newborn tissues, expression of the laminin alpha4 chain was mainly localized to mesenchymal cells. Strong expression was seen in the villi and submucosa of the developing intestine, the mesenchymal stroma surrounding the branching lung epithelia, and the external root sheath of vibrissae follicles, as well as in cardiac and skeletal muscle fibers. In the developing kidney, intense but transient expression was associated with the differentiation of epithelial kidney tubules from the nephrogenic mesenchyme. Immunohistologic staining with affinity-purified IgG localized the laminin alpha4 chain primarily to lung septa, heart, and skeletal muscle, capillaries, and perineurium. PMID- 9346929 TI - ATP depletion induces a loss of respiratory epithelium functional integrity and down-regulates CFTR (cystic fibrosis transmembrane conductance regulator) expression. AB - To mimic the effect of ischemia on the integrity of airway epithelium and expression of cystic fibrosis transmembrane conductance regulator (CFTR), we induced an ATP depletion of the respiratory epithelium from upper airway cells (nasal tissue) and human bronchial epithelial 16HBE14o- cell line. Histological analysis showed that 2 h of ATP depletion led to a loss of the epithelium integrity at the interface between basal cells and columnar cells. The expression of connexin 43 (Cx43, subunit of the gap junctions) and desmoplakins 1 and 2 (DPs 1 and 2, major components of the desmosomes) proteins was inhibited. After 90 min of ATP depletion, a significant decrease of the transepithelial resistance (25%) was observed but was reversible. Similar results were obtained with the 16HBE14o- human bronchial epithelial cell line. ATP depletion led to actin filaments depolymerization. The expression of the mature CFTR (170 kDa) and fodrin proteins at the apical domain of the ciliated cells was down-regulated. The steady-state levels of CFTR, Cx43, DPs 1 and 2 mRNAs, semiquantified by RT-polymerase chain reaction kinetics, remained constant throughout ATP depletion in nasal tissue as in the homogeneous cell population of 16HBE14o- human bronchial epithelial cell line. This suggests that the down-regulation of these proteins might be posttranscriptional. The intercellular diffusion through gap junctions of Lucifer dye was completely inhibited after 90 min of ATP depletion but was reversible. The volume-dependent and the cAMP-dependent chloride secretion were inhibited in a nonreversible way. Taken together, these results suggest that an ATP depletion in human airway epithelium, mimicking ischemia, may induce a marked alteration in the junctional complexes and cytoskeleton structure concomitantly with a loss of apical CFTR expression and chloride secretion function. PMID- 9346934 TI - Inhibition of sickle beta-chain (betaS)-dependent polymerization by nonhuman alpha-chains. A superinhibitory mouse-horse chimeric alpha-chain. AB - Horse alpha-chain inhibits sickle beta-chain-dependent polymerization; however, its inhibitory potential is not as high as that of mouse alpha-chain. Horse alpha (1-30) and alpha-(31-141) segments make, respectively, minor and major contributions to the inhibitory potential of horse alpha-chain. The sum of the inhibitory potential of the two segments does not account for the inhibitory potential of the full-length horse alpha-chain. Although the polymerization inhibitory potential of horse alpha-chain is lower than mouse alpha-chain, the inhibitory potential of horse alpha-(31-141) is comparable to that of mouse alpha (31-141). When mouse alpha-(1-30) is stitched to horse alpha-(31-141), the product is a chimeric alpha-chain with an inhibitory potential greater than mouse alpha-chain. In contrast, the stitching of horse alpha-(1-30) with mouse alpha (31-141) had no additional inhibitory potential. Molecular modeling studies of HbS containing the mouse-horse chimeric alpha-chain indicate altered side-chain interactions at the alpha1beta1 interface when compared with HbS. In addition, the AB/GH corner perturbations facilitate a different stereochemistry for the interaction of the epsilon-amino group of Lys-16(alpha) with the beta-carboxyl group of Asp-116(alpha), resulting in a decrease in the accessibility of the side chain of Lys-16(alpha) to the solvent. Based on molecular modeling, we speculate that these perturbations by themselves, or in synergy with the altered conformational aspects of the alpha1beta1 interactions, represent the molecular basis of the superinhibitory potential of the mouse-horse chimeric alpha-chains. PMID- 9346935 TI - Direct interaction of the KRAB/Cys2-His2 zinc finger protein ZNF74 with a hyperphosphorylated form of the RNA polymerase II largest subunit. AB - We previously identified ZNF74 as a developmentally expressed gene commonly deleted in DiGeorge syndrome. ZNF74 encodes an RNA-binding protein tightly associated with the nuclear matrix and belongs to a large subfamily of Cys2-His2 zinc finger proteins containing a KRAB (Kruppel-associated box) repressor motif. We now report on the multifunctionality of the zinc finger domain of ZNF74. This nucleic acid binding domain is shown here to function as a nuclear matrix targeting sequence and to be involved in protein-protein interaction. By far Western analysis and coimmunoprecipitation studies, we demonstrate that ZNF74 interacts, via its zinc finger domain, with the hyperphosphorylated largest subunit of RNA polymerase II (pol IIo) but not with the hypophosphorylated form. The importance of the phosphorylation in this interaction is supported by the observation that phosphatase treatment inhibits ZNF74 binding. Double immunofluorescence experiments indicate that ZNF74 colocalizes with the pol IIo and the SC35 splicing factor in irregularly shaped subnuclear domains. Thus, ZNF74 sublocalization in nuclear domains enriched in pre-mRNA maturating factors, its RNA binding activity, and its direct phosphodependent interaction with the pol IIo, a form of the RNA polymerase functionally associated with pre- mRNA processing, suggest a role for this member of the KRAB multifinger protein family in RNA processing. PMID- 9346936 TI - Crystal structure of rat Bcl-xL. Implications for the function of the Bcl-2 protein family. AB - Bcl-xL is a member of the Bcl-2 protein family, which regulates apoptosis. Preparation of recombinant rat Bcl-xL yielded two forms, one deamidated at -Asn Gly- sequences to produce isoaspartates and the other not deamidated. The crystal structures of the two forms show that they both adopt an essentially identical backbone structure which resembles the fold of human Bcl-xL: three layers of two alpha-helices each, capped at one end by two short helices. Both forms have a long disordered region, which contains the potential deamidation sites. The molecular structure exhibits a low level of interhelical interactions, the presence of three cavities, and a notable hydrophobic cleft surrounded by walls rich in basic residues. These unique structural features may be favorable for its accommodation into membranes or for possible rearrangement to modulate homo /heterodimerization. Homology modeling of Bcl-2 and Bax, based on the Bcl-xL structure, suggests that Bax has the strongest potential for membrane insertion. Furthermore, we found a possible interface for interaction with non-Bcl-2 family member proteins, such as CED-4 homologues. PMID- 9346937 TI - The fate of trichohyalin. Sequential post-translational modifications by peptidyl arginine deiminase and transglutaminases. AB - Trichohyalin (THH) is a major structural protein of the inner root sheath cells and medulla layer of the hair follicle and, to a lesser extent, of other specialized epithelia. THH is a high molecular weight insoluble alpha-helix-rich protein that forms rigid structures as a result of postsynthetic modifications by two Ca2+-dependent enzymes, transglutaminases (TGases) (protein cross-linking) and peptidyl-arginine deiminase (conversion of arginines to citrullines with loss of organized structure). The modified THH is thought to serve as a keratin intermediate filament matrix protein and/or as a constituent of the cell envelope. In this paper, we have explored in vitro the order of processing of THH to fulfill these functions, using an expressed truncated, more soluble form THH 8. THH-8 is a complete substrate for three known TGases expressed in epithelia, but the kinetic efficiency with TGase 3 is by far the greatest. Following maximal conversion of its arginines to citrullines, THH-8 is cross-linked even more efficiently by TGase 3, using most glutamines partially and all lysines. In addition, we show that insoluble aggregates of THH-8 or native pig tongue THH can be solubilized following peptidyl-arginine deiminase modification. Together, these data suggest an in vivo model in which THH located in insoluble cytoplasmic droplets is first modified by peptidyl-arginine deiminase which denatures it and makes it more soluble. This renders it available for efficient cross-linking by TGase 3 to form highly cross-linked rigid structures in the cells. This temporal order of reaction is supported by the observation that THH is expressed in hair follicle cells before the TGase 3 enzyme. PMID- 9346938 TI - Oncogenic c-Ki-ras but not oncogenic c-Ha-ras up-regulates CEA expression and disrupts basolateral polarity in colon epithelial cells. AB - Colon carcinomas commonly contain mutations in Ki-ras4B, but very rarely in Ha ras, suggesting that different Ras isoforms may have distinct functions in colon epithelial cell biology. In an earlier study we had demonstrated that oncogenic Ki-ras4BVal-12, but not oncogenic Ha-rasVal-12, blocks the apicobasal polarization of colon epithelial cells by preventing normal glycosylation of the integrin beta1 chain of the collagen receptor. As a result, only the Ki-ras mutated cells exhibited altered cell to substratum attachment, whereas mutation of either Ras isoform activated mitogen-activated protein kinases. We have now asked whether intercellular adhesion proteins implicated in establishing basolateral polarity in colon epithelial cells are modulated by oncogenic Ki Ras4BVal-12 proteins but not oncogenic Ha-RasVal-12 proteins. The embryonic adhesion protein carcinoembryonic antigen (CEA) was up-regulated on the mRNA and protein levels in each of three stable Ki-rasVal-12 transfectant lines but in none of three stable Ha-rasVal-12 transfectant lines. The elevated protein levels of CEA in Ki-ras4BVal-12 transfectant cells were decreased by blocking expression of Ki-ras4BVal-12 with antisense oligonucleotides. N-cadherin levels were decreased in only the Ki-ras transfectants, whereas E-cadherin levels were unchanged. Immunohistochemical analysis demonstrated that Ki-ras4BVal-12 transfectant cells did not polarize into cells with discrete apical and basal regions and so could not restrict expression of CEA to the apical region. These unpolarized cells displayed elevated levels of CEA all along their surface membrane where CEA mediated random, multilayered associations of tumor cells. This aggregation was both calcium-independent and blocked by Fab' fragments of anti-CEA monoclonal antibody col-1. Trafficking of the lysosomal cysteine protease cathepsin B may also be altered when cell polarity cannot be established. Ki-ras4BVal-12 transfectant cells expressed 2-fold elevated protein levels of the lysosomal cysteine protease cathepsin B but did not up-regulate cathepsin B mRNA expression. One function of oncogenic c-Ki-Ras proteins in colon cancer progression may be to up-regulate CEA and thus to prevent the lateral adhesion of adjacent colon epithelial cells that normally form a monolayer in vivo. PMID- 9346939 TI - The Kruppel-associated box (KRAB)-zinc finger protein Kid-1 and the Wilms' tumor protein WT1, two transcriptional repressor proteins, bind to heteroduplex DNA. AB - Zinc finger proteins of the Cys2His2 class represent a large group of DNA-binding proteins. A major subfamily of those proteins, the Kruppel-associated box (KRAB) domain-containing Cys2His2-zinc finger proteins, have been described as potent transcriptional repressors. So far, however, no DNA-binding sites for KRAB domain containing zinc finger proteins have been isolated. Using a polymerase chain reaction-based selection strategy with double- and single-stranded DNA, we failed to reveal a binding site for Kid-1, one member of KRAB-zinc finger proteins. Binding of Kid-1 both to single- and homoduplex double-stranded DNA was negligible. We now present evidence that Kid-1 binds to heteroduplex DNA. Similar to Kid-1, the non-KRAB-zinc finger protein WT1 also bound avidly to heteroduplex DNA (both the -KTS and +KTS splice variant of WT1), whereas the POU domain protein Oct-6, the ets domain protein Ets-1 and the RING finger of BRCA-1 did not bind to heteroduplex DNA. Binding of WT1 to heteroduplex DNA was markedly reduced in naturally occurring mutants. The recognition of certain DNA structures by transcriptional repressor proteins may therefore represent a more common phenomenon than previously thought. PMID- 9346940 TI - Interaction between the adhesion receptor, CD44, and the oncogene product, p185HER2, promotes human ovarian tumor cell activation. AB - In this study we have examined the interaction between CD44s (the standard form) and the p185(HER2) proto-oncogene in the ovarian carcinoma cell line. Surface biotinylation followed by wheat germ agglutinin column chromatography and anti CD44-mediated immunoprecipitation indicate that both CD44s and p185(HER2) are expressed on the cell surface and most importantly, that these two molecules are physically linked to each other via interchain disulfide bonds. We have also determined that hyaluronic acid stimulates CD44s-associated p185(HER2) tyrosine kinase activity, leading to an increase in the ovarian carcinoma cell growth. After transfection of the ovarian carcinoma cell line with the adenovirus 5 E1A gene, which is known to repress p185(HER2) expression, we observed that both surface CD44s expression and CD44s-mediated cell adhesion to hyaluronic acid are significantly reduced in the transfectant cells compared with the control cells. These data suggest that down-regulation of p185(HER2) blocks CD44s expression and subsequent adhesion function. Our findings also indicate that the CD44s p185(HER2) interaction is both functionally coupled and biosynthetically regulated. We believe that direct "cross-talk" between these two surface molecules (i.e. CD44s and the p185(HER2)) may be one of the most important signaling events in human ovarian carcinoma development. PMID- 9346941 TI - Fidelity of Escherichia coli DNA polymerase III holoenzyme. The effects of beta, gamma complex processivity proteins and epsilon proofreading exonuclease on nucleotide misincorporation efficiencies. AB - The fidelity of Escherichia coli DNA polymerase III (pol III) is measured and the effects of beta, gamma processivity and epsilon proofreading subunits are evaluated using a gel kinetic assay. Pol III holoenzyme synthesizes DNA with extremely high fidelity, misincorporating dTMP, dAMP, and dGMP opposite a template G target with efficiencies finc = 5.6 x 10(-6), 4.2 x 10(-7), and 7 x 10(-7), respectively. Elevated dGMP.G and dTMP.G misincorporation efficiencies of 3.2 x 10(-5) and 5.8 x 10(-4), attributed to a "dNTP-stabilized" DNA misalignment mechanism, occur when C and A, respectively, are located one base downstream from the template target G. At least 92% of misinserted nucleotides are excised by pol III holoenzyme in the absence of a next correct "rescue" nucleotide. As rescue dNTP concentrations are increased, pol III holoenzyme suffers a maximum 8-fold reduction in fidelity as proofreading of mispaired primer termini are reduced in competition with incorporation of a next correct nucleotide. Compared with pol III holoenzyme, the alpha holoenzyme, which cannot proofread, has 47-, 32-, and 13-fold higher misincorporation rates for dGMP.G, dTMP.G, and dAMP.G mispairs. Both the beta, gamma complex and the downstream nucleotide have little effect on the fidelity of catalytic alpha subunit. An analysis of the gel kinetic fidelity assay when multiple polymerase-DNA encounters occur is presented in the "Appendix" (see Fygenson, D. K., and Goodman, M. F. (1997) J. Biol. Chem. 272, 27931-27935 (accompanying paper)). PMID- 9346942 TI - Appendix. Gel kinetic analysis of polymerase fidelity in the presence of multiple enzyme DNA encounters. PMID- 9346943 TI - Binding sites and binding properties of binary and ternary complexes of insulin like growth factor-II (IGF-II), IGF-binding protein-3, and acid-labile subunit. AB - We have examined regions of rat IGF-binding protein-3 (IGFBP-3) important for complex formations using two kinds of deletion mutants, three kinds of chimera molecules between rat IGFBP-3 and rat IGFBP-2, and a synthetic peptide (41 residues, Glu52-Ala92) derived from rat IGFBP-3. Solid-phase binding assays using 96-well microtiter plates were designed to quantitate the relative binding affinities. It was found that not only the IGFBP-3 derivatives with the amino terminal, cysteine-rich domain (N domain) but also the synthetic peptide maintained affinity for IGF-II. Ternary complex formation was observed with full length IGFBP-3 and chimera IGFBP, the carboxyl-terminal cysteine-rich domain (C domain) of which was derived from IGFBP-3, unlike the mutants lacking the C domain and the chimera IGFBPs, the C domain of which was derived from IGFBP-2. These results were confirmed by affinity cross-linking experiments. Furthermore, the IGFBP-3 derivatives that possessed the C domain of IGFBP-3 bound to the acid labile subunit, even in the absence of IGFs. Finally, we observed sites in IGF-II important for the ternary complex formation using various IGF-II mutants. These IGF-II mutants, which contained a substitution of Tyr27 for Leu, had extremely reduced activity. These results strongly suggest that: 1) the N domain, containing at least Glu52-Ala92, of rat IGFBP-3 is important for binding to IGF II; 2) the C domain of IGFBP-3 is essential for binding to the acid-labile subunit both in the presence and absence of IGF-II; and 3) Tyr27 of IGF-II is important for the ternary complex formation. PMID- 9346944 TI - Phage display selection on whole cells yields a peptide specific for melanocortin receptor 1. AB - A phage display system for the selection of peptides binding to heterologously expressed human melanocortin receptor 1 on the surface of insect cells has been established. It could be shown that phage particles displaying the natural ligand alpha-melanocyte-stimulating hormone bind selectively to cells expressing this receptor and that these phages exhibit biological activity on mouse B16F1 melanoma cells. Insect cells were superior to other cell lines tested and have been used to select binders from a small library, in which critical determinants (Phe7-Arg8-Trp9) were kept, whereas the flanking regions where allowed to variate freely. One peptide displaying little similarity with native hormone was found that binds to the receptor also in its free form with an affinity of 7 nM. It showed a remarkable selectivity for this receptor, because it binds to the other melanocortin receptor subtypes with a maximum affinity of 21 microM. This is the first time phage display has been used successfully with G-protein-coupled receptors lacking an extracellular binding domain. PMID- 9346945 TI - Dimerizing the estrogen receptor DNA binding domain enhances binding to estrogen response elements. AB - In this work, we provide a rationale for the finding that the estrogen receptor (ER) binds to its DNA response element as a homodimer in vivo. Binding of the monomer estrogen receptor DNA binding domain (ER DBD) to a palindromic, consensus estrogen response element (ERE) is increased 5-6-fold when the ER DBD is dimerized either by a monoclonal antibody that recognizes an attached epitope tag or by expressing the ER DBD as a single molecule in which the two monomers are joined by a peptide linker. Most of the increase in binding is due to stabilization of the ER DBD.ERE complex. We observed only an approximately 2.5 fold reduction in binding when a consensus ERE was replaced with widely spaced ERE half-sites, suggesting that the interaction between ER DBDs on the ERE is relatively weak, and that in full-length ER the DBDs can move independently of each other. To test binding to an imperfect palindrome, typical of the imperfect EREs found in almost all natural estrogen receptor responsive genes, we used the pS2 ERE. Even at high concentrations of ER DBD, specific binding of the ER DBD to the imperfect pS2 ERE was undetectable. Both of the dimerized ER DBDs exhibited efficient binding to the imperfect pS2 ERE, with an affinity at least 25-fold greater than monomer ER DBD. These data support the view that steroid receptor dimerization provides an important mechanism facilitating the recognition of naturally occurring, imperfect hormone response elements. PMID- 9346946 TI - Identification of an upstream enhancer within a functional promoter of the human leukemia inhibitory factor receptor gene and its alternative promoter usage. AB - Knockout of the leukemia inhibitory factor receptor (LIFR) gene results in disrupted placental architecture, imbalanced bone development, and losses of functional neurons. We here report the identification of an enhancer in a functional human LIFR gene promoter and alternative promoter usage by this gene. A single transcription start site was identified in placental JEG-3 cells and a genomic clone containing 4876-nucleotide upstream sequences was found to have promoter activity in JEG-3 cells. However, in osteogenic sarcoma U-2 OS cell, Northern blot using a probe of the first exon detected in JEG-3 cells failed to detect LIFR transcripts. 5'-Rapid amplification of cDNA ends (RACE) revealed an alternative first exon and a 0.6-kilobase pair (kb) 5'-flanking region possessed promoter activity in U-2 OS cells. For the 4.8-kb promoter active in placental cells, a minimal promoter was localized within -162 nucleotides. Three regions increased and one inhibited promoter activity. Subcloning of an activation region (-4876 to -3453 nucleotides) into SV40 promoter either upstream or downstream in either orientation to the luciferase reporter resulted in 10-35-fold luciferase induction, demonstrating the characteristics of an enhancer. Transfections into nine cell lines of different tissue origin indicated that the cloned promoter and enhancer in the 4.8-kb fragment was placental tissue-specific. A 226-base pair fragment (-4625 to -4400 nucleotides) was further localized as the minimal enhancer region, in which deletion of either element A (-4625 to -4581 nucleotides) or element B (-4418 to -4400 nucleotides) resulted in the loss of enhancer activity. Electrophoretic mobility shift assay confirmed that these two elements bind to specific nuclear proteins individually. In the middle region between element A and B, disruption of enhancer integrity also led to a loss of enhancer activity, although two SP1 and three NF-kappaB/c-Rel binding sites did not contribute to enhancer function. These results demonstrate a complex regulation of the human LIFR gene, including alternative promoter usage and tissue-specific elements at the transcription level. PMID- 9346947 TI - Alterations in human glomerular epithelial cells interacting with nonenzymatically glycosylated matrix. AB - The glomerular epithelial cells and the glomerular basement membrane are important constituents of the permselective barrier in the kidney. These are affected in diabetic nephropathy, one of the long-term complications in diabetic patients. Nonenzymatic glycosylation resulting in the accumulation of advanced glycosylation end products correlates with the development of long-term complications in diabetes. The interaction of cells with extracellular matrix proteins plays a critical role in a variety of biological processes. Recent studies show that cell-matrix interactions mediated by integrins can transduce biochemical signals to the cell interior and regulate cell behavior. In this paper we demonstrate that interactions of human glomerular epithelial cells with a nonenzymatically glycated matrix are altered with defective cell spreading, reduced phosphorylation of focal adhesion kinase and reduced activity of mitogen activated protein kinase. These data suggest that matrix glycation interferes with normal cell-matrix interactions and intracellular signaling that can potentially result in differential gene expression contributing to the changes seen in diabetic nephropathy. PMID- 9346948 TI - The Cys6 intermolecular disulfide bond and the collagen-like region of rat SP-A play critical roles in interactions with alveolar type II cells and surfactant lipids. AB - Rat pulmonary surfactant protein A is an oligomer of 18 polypeptide chains which are associated by triple helix formation in the collagen-like domain and interchain disulfide bridges at the NH2 terminus. The roles of the intermolecular bond at Cys6 and the collagen-like domain (Gly8-Pro80) in the interactions of SP A with phospholipids and alveolar type II cells were investigated using mutant forms of the protein. Wild type SP-A (SP-Ahyp), SP-A with the substitution Cys6 - > Ser to prevent disulfide formation (SP-Ahyp, C6S), and SP-A with the collagen domain deleted (SP-ADeltaG8-P80) were synthesized in insect cells using recombinant baculoviruses. The SP-As were glycosylated and secreted from the invertebrate cells and the binding affinities of the wild type and mutant proteins for the mannose-Sepharose matrix used for purification were nearly identical. The SP-Ahyp and SP-ADeltaG8-P80 were at least nonameric in solution based on gel exclusion chromatography, and demonstrated extensive sulfhydryl dependent oligomerization under nonreducing conditions. The SP-Ahyp,C6S was also oligomeric in solution and formed disulfide-dependent dimers, indicating the presence of at least one additional interchain disulfide bond. The SPADeltaG8-P80 but not the SP-Ahyp,C6S aggregated lipid vesicles at 20 degrees C and augmented the surface tension lowering effect of extracts of natural surfactant. The SP ADeltaG8-P80 competed poorly with native SP-A for receptor occupancy on isolated alveolar type II cells and was a potent but nonspecific (concanavalin A-like) inhibitor of surfactant secretion. In contrast, the SP-Ahyp,C6S partially competed for receptor occupancy and weakly inhibited surfactant secretion in a specific manner. Neither the SP-ADeltaG8-P80 nor the SP-Ahyp,C6S supported the association of phospholipid liposomes with type II cells. We conclude that: 1) the Cys6 interchain disulfide bond of SP-A is required for aggregation of liposomes and for potent inhibition of surfactant secretion. 2) The collagen-like region is required for competition with 125I-SP-A for receptor occupancy and specific inhibition of surfactant secretion in the presence of competing sugars. 3) Both the NH2-terminal disulfide and the collagen-like region are required to enhance the association of phospholipid vesicles with type II cells. PMID- 9346949 TI - Selective targeting and inhibition of yeast RNA polymerase II by RNA aptamers. AB - To probe the complex nucleic acid binding domains of yeast RNA polymerase II (Pol II), we have isolated in the presence of heparin RNA molecules that selectively bind to yeast Pol II. A class of RNA molecules was found to bind and strongly interfere with enzyme-DNA interaction but not with RNA chain elongation. Remarkably, one selected RNA ligand was a specific inhibitor of Saccharomyces cerevisiae Pol II. S. cerevisiae Pol I and Pol III and Pol II from Schizosaccharomyces pombe or wheat germ cells were not affected. Photocross linking experiments showed that the RNA ligand preferentially interacted with B220, the largest subunit of Pol II and, to a lesser extent, with B150, the second largest subunit. The selected RNA was expressed in yeast cells under the control of a Pol III promoter. Yeast cells that expressed the anti-Pol II aptamer grew normally. However, a cell growth defect was observed when expressing the RNA aptamer in cells having an artificially reduced level of Pol II. PMID- 9346950 TI - Role of Egr-1 gene expression in B cell receptor-induced apoptosis in an immature B cell lymphoma. AB - Ligation of B cell receptor (BCR) on BKS-2, an immature B cell lymphoma by anti IgM antibodies (Ab) caused apoptosis. Here we report that signaling through B cell receptor in wild type BKS-2 cells down-regulated the expression of Egr-1, a zinc finger-containing transcription factor. A reduction in the level of Egr-1 mRNA could be demonstrated as early as 30 min after the ligation of BCR on BKS-2 cells. Immunocytochemical and Western blot analysis revealed that the expression of EGR-1 protein was also inhibited by anti-IgM treatment. Antisense oligonucleotides to Egr-1 caused growth inhibition and apoptosis in BKS-2 cells, suggesting that expression of Egr-1 is important for the survival of these B lymphoma cells. In contrast to wild type BKS-2 cells, the mutant 1. B5 cell line, which is refractory to B cell receptor-mediated growth-inhibitory signals, showed an increased expression of Egr-1 upon treatment with anti-IgM. These results implicate a role for Egr-1 in blocking B cell receptor-mediated apoptosis in immature B cells. PMID- 9346951 TI - Relative functions of the alpha and beta subunits of the proteasome activator, PA28. AB - PA28 is a 180,000-dalton protein that activates hydrolysis of small nonubiquitinated peptides by the 20 S proteasome. PA28 is composed of two homologous subunits, alpha and beta, arranged in alternating positions in a ring shaped oligomer with a likely stoichiometry of (alphabeta)3. Our previous work demonstrated that the carboxyl terminus of the alpha subunit was necessary for PA28 to bind to and activate the proteasome. The goals of this work were to define the exact structural basis for this effect and to determine the relative roles of the alpha and beta subunits in proteasome activation. Each subunit and various mutants of the alpha subunit were expressed in Escherichia coli and purified. PA28alpha stimulated the proteasome, but had a much greater Kact than native heteromeric PA28. In contrast, PA28beta was unable to stimulate the proteasome. Mutants of the alpha subunit in which the carboxyl-terminal tyrosine residue was deleted or substituted with charged amino acids could neither bind to nor activate the proteasome. However, substitution of the carboxyl-terminal tyrosine with other amino acids resulted in proteins which could stimulate the proteasome to various extents. Tryptophan mutants stimulated the proteasome as well as did native PA28, whereas serine or phenylalanine mutants stimulated the proteasome much poorer than did wild type PA28alpha. Deletion of the "KEKE" motif, a 28-amino acid domain near the amino terminus of PA28alpha, had no effect on proteasome stimulatory activity. Hetero-oligomeric PA28 proteins were reconstituted from isolated wild type and mutant subunits. PA28 reconstituted from wild type subunits had structural and functional properties that were indistinguishable from those of the native hetero-oligomeric protein. PA28 molecules reconstituted from inactive alpha subunits and wild type beta subunits remained inactive. However, PA28 molecules reconstituted from suboptimally active alpha mutants and wild type beta subunits had the same activity as native heteromeric PA28. These results indicate that the beta subunit modulates PA28 activity, perhaps by influencing the affinity of PA28 for the proteasome. PMID- 9346952 TI - Isolation and characterization of cDNAs corresponding to an additional member of the human histone deacetylase gene family. AB - Several human cDNAs encoding a histone deacetylase protein, HDAC3, have been isolated. Analysis of the predicted amino acid sequence of HDAC3 revealed an open reading frame of 428 amino acids with a predicted molecular mass of 49 kDa. The HDAC3 protein is 50% identical in DNA sequence and 53% identical in protein sequence compared with the previously cloned human HDAC1. Comparison of the HDAC3 sequence with human HDAC2 also yielded similar results, with 51% identity in DNA sequence and 52% identity in protein sequence. The expressed HDAC3 protein is functionally active because it possesses histone deacetylase activity, represses transcription when tethered to a promoter, and binds transcription factor YY1. Similar to HDAC1 and HDAC2, HDAC3 is ubiquitously expressed in many different cell types. PMID- 9346953 TI - Molecular cloning and expression of mouse and human cDNAs encoding heparan sulfate D-glucosaminyl 3-O-sulfotransferase. AB - The cellular rate of anticoagulant heparan sulfate proteoglycan (HSPGact) generation is determined by the level of a kinetically limiting microsomal activity, HSact conversion activity, which is predominantly composed of the long sought heparan sulfate D-glucosaminyl 3-O-sulfotransferase (3-OST) (Shworak, N. W., Fritze, L. M. S., Liu, J., Butler, L. D., and Rosenberg, R. D. (1996) J. Biol. Chem. 271, 27063-27071; Liu, J., Shworak, N. W., Fritze, L. M. S., Edelberg, J. M., and Rosenberg, R. D. (1996) J. Biol. Chem. 271, 27072-27082). Mouse 3-OST cDNAs were isolated by proteolyzing the purified enzyme with Lys-C, sequencing the resultant peptides as well as the existing amino terminus, employing degenerate polymerase chain reaction primers corresponding to the sequences of the peptides as well as the amino terminus to amplify a fragment from LTA cDNA, and utilizing the resultant probe to obtain full-length enzyme cDNAs from a lambda Zap Express LTA cDNA library. Human 3-OST cDNAs were isolated by searching the expressed sequence tag data bank with the mouse sequence, identifying a partial-length human cDNA and utilizing the clone as a probe to isolate a full-length enzyme cDNA from a lambda TriplEx human brain cDNA library. The expression of wild-type mouse 3-OST as well as protein A-tagged mouse enzyme by transient transfection of COS-7 cells and the expression of both wild-type mouse and human 3-OST by in vitro transcription/translation demonstrate that the two cDNAs directly encode both HSact conversion and 3-OST activities. The mouse 3 OST cDNAs exhibit three different size classes because of a 5'-untranslated region of variable length, which results from the insertion of 0-1629 base pairs (bp) between residues 216 and 217; however, all cDNAs contain the same open reading frame of 933 bp. The length of the 3'-untranslated region ranges from 301 to 430 bp. The nucleic acid sequence of mouse and human 3-OST cDNAs are approximately 85% similar, encoding novel 311- and 307-amino acid proteins of 35,876 and 35,750 daltons, respectively, that are 93% similar. The encoded enzymes are predicted to be intraluminal Golgi residents, presumably interacting via their C-terminal regions with an integral membrane protein. Both 3-OST species exhibit five potential N-glycosylation sites, which account for the apparent discrepancy between the molecular masses of the encoded enzyme (approximately 34 kDa) and the previously purified enzyme (approximately 46 kDa). The two 3-OST species also exhibit approximately 50% similarity with all previously identified forms of the heparan biosynthetic enzyme N-deacetylase/N sulfotransferase, which suggests that heparan biosynthetic enzymes share a common sulfotransferase domain. PMID- 9346954 TI - In situ compositional analysis of acidocalcisomes in Trypanosoma cruzi. AB - We measured the elemental content of different compartments in Trypanosoma cruzi epimastigotes using quick freezing, ultracryomicrotomy, and electron probe microanalysis. Vacuoles identified by high electron density contained (in units of mmol/kg dry weight +/- S.E.) large amounts of phosphorus (1390 +/- 13), magnesium (646 +/- 19), calcium (171 +/- 5), sodium (161 +/- 18), and zinc (148 +/- 6). No other compartment had appreciable calcium or zinc content. Iron (128 +/- 16 mmol/kg) was detected only in vacuoles distinct from the electron-dense vacuoles and other organelles. Incubation of cells for 70 min in culture medium in the presence of ionomycin plus nigericin led to a very significant 3- or 2 fold increase in potassium in the electron-dense vacuoles and the iron-rich vacuoles, respectively, with no significant change in the other elements investigated. This indicated the acidic nature of the vacuoles and demonstrated that the electron-dense vacuoles correspond to what were described previously as acidocalcisomes, i.e. acidic compartments rich in Ca2+. The acidocalcisomes were investigated by separation of epimastigote fractions on Percoll gradients in combination with Triton WR-1339 treatment. This detergent caused a rapid vacuolation; these vacuoles were shown by electron microscopy to be largely transparent, with a diffuse matrix. Percoll gradient fractionation demonstrated decreases in the density of various organelle markers in detergent-treated cells compared with controls. Large decreases in the density of the acidocalcisome and the mitochondrion were seen, as well as smaller decreases in the density of the other markers. Conventional electron microscopy of epimastigotes loaded with gold labeled transferrin indicated that the endosomal system was separate from vacuoles that probably corresponded to the calcium-containing organelles detected by electron probe microanalysis. The combined results provide evidence that acidocalcisomes are organelles different from lysosomes or other organelles previously described in these parasites. PMID- 9346955 TI - Dynamin assembles into spirals under physiological salt conditions upon the addition of GDP and gamma-phosphate analogues. AB - Dynamin is a 100-kDa GTPase that is believed to be involved in the constriction of clathrin-coated pits and the fission of clathrin-coated vesicles during receptor-mediated endocytosis and during membrane retrieval in nerve termini. It has been shown that purified dynamin incubated under low salt conditions forms rings and spirals that, in dimension and appearance, resemble the dense material occasionally observed at the necks of coated pits. In this report we show that purified dynamin forms spirals under physiological salt conditions when incubated with GDP and gamma-phosphate analogues (beryllium and aluminum fluoride) or when dialyzed into guanosine 5'-3-O-(thio)triphosphate. Moreover, spirals still form when dynamin is proteolyzed to either a predominant approximately 90-kDa species, lacking the C terminus, or to two smaller fragments, a approximately 55-kDa species originating from the N-terminal half of the protein and a approximately 30-kDa species lacking both the N and C termini. This work indicates that the addition of GDP and gamma-phosphate analogues arrests dynamin in a GTP or transition state that markedly stabilizes the spiral conformation under physiological ionic strength conditions and thereby suggests that dynamin in the absence of a receptor is capable of assembly into spirals at the necks of coated pits prior to vesicle fission. PMID- 9346956 TI - Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation. AB - SNAP-25, syntaxin, and synaptobrevin play a key role in the regulated exocytosis of synaptic vesicles, but their mechanism of action is not understood. In vitro, the proteins spontaneously assemble into a ternary complex that can be dissociated by the ATPase N-ethylmaleimide-sensitive fusion protein and the cofactors alpha-, beta-, and gamma-SNAP. Since the structural changes associated with these reactions probably form the basis of membrane fusion, we have embarked on biophysical studies aimed at elucidating such changes in vitro using recombinant proteins. All proteins were purified in a monomeric form. Syntaxin showed significant alpha-helicity, whereas SNAP-25 and synaptobrevin exhibited characteristics of largely unstructured proteins. Formation of the ternary complex induced dramatic increases in alpha-helicity and in thermal stability. This suggests that structure is induced in SNAP-25 and synaptobrevin upon complex formation. In addition, the stoichiometry changed from 2:1 in the syntaxin-SNAP 25 complex to 1:1:1 in the ternary complex. We propose that the transition from largely unstructured monomers to a tightly packed, energetically favored ternary complex connecting two membranes is a key step in overcoming energy barriers for membrane fusion. PMID- 9346957 TI - The 66-kDa Shc isoform is a negative regulator of the epidermal growth factor stimulated mitogen-activated protein kinase pathway. AB - In addition to tyrosine phosphorylation of the 66-, 52-, and 46-kDa Shc isoforms, epidermal growth factor (EGF) treatment of Chinese hamster ovary cells expressing the human EGF receptor also resulted in the serine/threonine phosphorylation of approximately 50% of the 66-kDa Shc proteins. The serine/threonine phosphorylation occurred subsequent to tyrosine phosphorylation and was prevented by pretreatment of the cells with the MEK-specific inhibitor PD98059. Surprisingly, only the gel-shifted 66-kDa Shc isoform (serine/threonine phosphorylated) was tyrosine phosphorylated and associated with Grb2. In contrast, only the non-serine/threonine-phosphorylated fraction of 66-kDa Shc was associated with the EGF receptor. To assess the relationship between the three Shc isoforms in EGF-stimulated signaling, the cDNA encoding the 66-kDa Shc species was cloned from a 16-day-old mouse embryo library. Sequence alignment confirmed that the 66-kDa Shc cDNA resulted from alternative splicing of the primary Shc transcript generating a 110-amino acid extension at the amino terminus. Co-immunoprecipitation of Shc and Grb2 from cells overexpressing the 52/46-kDa Shc isoforms versus the 66-kDa Shc species directly demonstrated a competition of binding for a limited pool of Grb2 proteins. Furthermore, expression of the 66-kDa Shc isoform markedly accelerated the inactivation of ERK following EGF stimulation. Together, these data indicate that the serine/threonine phosphorylation of 66-kDa Shc impairs its ability to associate with the tyrosine-phosphorylated EGF receptor and can function in a dominant interfering manner by inhibiting EGF receptor downstream signaling pathways. PMID- 9346958 TI - Mechanical strain increases expression of the brain natriuretic peptide gene in rat cardiac myocytes. AB - Using a device that applies cyclical strain (1 Hz) to ventricular cardiocytes cultured on collagen-coated silicone elastomer surfaces, we have demonstrated strain-dependent increases in brain natriuretic peptide (BNP) secretion, BNP mRNA levels, and expression of a transiently transfected -1595 human BNP-luciferase reporter. When actinomycin D (10 microM) was introduced concomitantly with the strain stimulus, the strain-induced increase in BNP mRNA was eliminated, and the decay of transcripts was identical in the control and strained cells, indicating the lack of independent effects on transcript stability. Strain-dependent -1595 human BNP-luciferase activity was completely inhibited by chelerythrine, 2 aminopurine, genistein, and W-7 and only partially or not at all by KN-62, wortmannin, and H-89. The effects of these individual agents paralleled their effects on mitogen-activated protein kinase (MAPK) activity, but not c-Jun N terminal kinase (JNK) activity, in the cells. Overexpression of wild-type MAPK and, to a lesser extent, JNK increased strain-dependent BNP promoter activity, whereas dominant-negative mutants of MAPK kinase, JNK kinase, or Ras completely blocked strain-dependent reporter activity. These findings provide the first demonstration that mechanical strain can increase myocardial gene expression through a transcriptional mechanism and suggest important roles for MAPK and JNK in mediating this effect. PMID- 9346959 TI - Identification of hyaluronan-binding domains of aggrecan. AB - Aggrecan, a large cartilage proteoglycan, interacts with hyaluronan (HA), to form aggregates which function to resist compression in joints. The N-terminal region of aggrecan contains two structurally related globular domains, G1 and G2 separated by IGD domain. The G1 domain consists of three subdomains, A, B, and B', structural features characteristic to many other HA-binding proteoglycans. Here, we studied the interaction of aggrecan domains with HA using recombinant proteins expressed in 293 cells, an embryonal kidney cell line. Deglycosylation of the recombinant aggrecan fragment reduced the HA binding activity. We found that both the B and B' subdomains were required for HA binding and that a single module of A, B, or B' was unable to bind HA. The A subdomain increased the HA binding activity of the B-B' region. The G2 domain had no HA binding activity confirming previous reports. Studies of HA-binding properties using a BIAcoreTM biosensor system revealed that the KD of recombinant aggrecan fragment (AgW) consisting of G1, IGD, and G2 was 0.226 microM, whereas the KD of another HA binding protein, native bovine link protein, is 0.089 microM. In contrast, AgMut11 which lacked subdomain A showed little HA binding activity. AgMut12 consisting of only B-B' had a 3.4-fold lower affinity and AgMut13 containing A-B B' was 1.5-fold lower than AgW. These results suggest that carbohydrates are essential for high level aggrecan binding to HA and that the A subdomain of aggrecan functions in a cooperative manner with subdomains B and B'. PMID- 9346960 TI - Molecular cloning and characterization of 12-oxophytodienoate reductase, an enzyme of the octadecanoid signaling pathway from Arabidopsis thaliana. Structural and functional relationship to yeast old yellow enzyme. AB - Using partial amino acid sequence information for 12-oxophytodienoate-10,11 reductase obtained from Corydalis sempervirens we have cloned the homologous enzyme from Arabidopsis thaliana. The open reading frame of the cDNA encodes a polypeptide of 372 amino acids (Mr = 41,165) with significant similarity to the sequence of Old Yellow Enzyme from Saccharomyces carlsbergensis (Saito, K., Thiele, D. J., Davio, M., Lockridge, O., and Massey, V. (1991) J. Biol. Chem. 266, 20720-20724), a flavin (FMN)-protein catalyzing the NADPH-dependent reduction of the olefinic bond of alpha,beta-unsaturated carbonyls. Specifically, all residues required for binding of FMN in Old Yellow Enzyme are conserved in the A. thaliana sequence, as are all residues associated with catalytic activity. The enzyme was functionally expressed from its cDNA in Escherichia coli and thus proven to encode OPDA reductase. Further similarities of OPDA reductase and yeast Old Yellow Enzyme include their binding to and elution by reductant from N-(4 hydroxybenzoyl)aminohexyl-Sepharose, the immunoreactivity of yeast Old Yellow Enzyme with an antiserum raised against plant OPDA reductase and the demonstration that Old Yellow Enzyme is an active OPDA reductase. It is thus conceivable that the physiological role of Old Yellow Enzymes now known from bacteria, yeasts, and higher plants, is in oxylipin metabolism. PMID- 9346961 TI - Expression of wild-type p53 is required for efficient global genomic nucleotide excision repair in UV-irradiated human fibroblasts. AB - We have shown previously that Li-Fraumeni syndrome fibroblasts homozygous for p53 mutations are deficient in the removal of UV-induced cyclobutane pyrimidine dimers from genomic DNA, but still proficient in the transcription-coupled repair pathway (Ford, J. M., and Hanawalt, P. C. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 8876-8880). We have now utilized monoclonal antibodies specific for cyclobutane pyrimidine dimers or 6-4 photoproducts, respectively, to measure their repair in UV-irradiated human fibroblasts. Cells homozygous for p53 mutations were deficient in the repair of both photoproducts, whereas cells heterozygous for mutant p53 exhibited normal repair of 6-4 photoproducts, but decreased initial rates of removal of cyclobutane pyrimidine dimers, compared with normal cells. The specificity of the effect of wild-type p53 on nucleotide excision repair was demonstrated in a p53 homozygous mutant cell line containing a tetracycline-regulated wild-type p53 gene. Wild-type p53 expression and activity were suppressed in the presence of tetracycline, whereas withdrawal of tetracycline resulted in the induction of p53 expression, cell cycle checkpoint activation, and DNA damage-induced apoptosis. The regulated expression of wild type p53 resulted in the recovery of normal levels of repair of both cyclobutane pyrimidine dimers and 6-4 photoproducts in genomic DNA, but did not alter the transcription-coupled repair of cyclobutane pyrimidine dimers. Therefore, the wild-type p53 gene product is an important determinant of nucleotide excision repair activity in human cells. PMID- 9346962 TI - Human SPA-1 gene product selectively expressed in lymphoid tissues is a specific GTPase-activating protein for Rap1 and Rap2. Segregate expression profiles from a rap1GAP gene product. AB - Mouse Spa-1 gene with a region homologous to the human rap1GAP gene is transcriptionally induced in the lymphocytes by mitogenic stimulation. Herein we have cloned a cDNA for its human counterpart. SPA-1 cDNA encodes a 130-kDa protein (p130(SPA-1)) consisting of proline-rich regions and rap1GAP-related domain followed by a coiled-coil stretch. Baculovirally expressed p130(SPA-1) exhibited GTPase-activating protein (GAP) activity for Rap1 and Rap2, but not for Ras, Rho, Cdc42, Rac, and Ran, with comparable specific activity to the rap1GAP gene product (p85/95(rap1GAP)). In the cells, p130(SPA-1) was mostly localized at the perinuclear membranous region co-localizing with Rap1 and Rap2. Expression of SPA-1 and rap1GAP genes tended to be segregate in various tissues, lymphoid tissues expressing abundant SPA-1 transcript without rap1GAP, while those such as brain, kidney, and pancreas exhibiting rap1GAP mRNA with little SPA-1. Promyelocytic HL-60 cells, which expressed p130(SPA-1) with little p85/95(rap1GAP) in uninduced state, showed progressive decline in p130(SPA-1) and conversely drastic increase in p85/95(rap1GAP) as they ceased from proliferation and differentiated into macrophages by 12-O-tetradecanoylphorbol-13-acetate. These results suggested that products of SPA-1 and rap1GAP genes, albeit comparable GAP activity for Rap1 and Rap2, functioned in the distinct contexts depending on cell types and/or states. PMID- 9346963 TI - Mimecan, the 25-kDa corneal keratan sulfate proteoglycan, is a product of the gene producing osteoglycin. AB - Bovine cornea contains three unique keratan sulfate proteoglycans (KSPGs), of which two (lumican and keratocan) have been characterized using molecular cloning. The gene for the third protein (KSPG25) has not been identified. This study examined the relationship between the KSPG25 protein and the gene for osteoglycin, a 12-kDa bone glycoprotein. The N-terminal amino acid sequence of KSPG25 occurs in osteoglycin cDNA cloned from bovine cornea. The osteoglycin amino acid sequence makes up the C-terminal 47% of the deduced sequence of the KSPG25 protein. Antibodies to osteoglycin reacted with intact corneal KSPG, with KSPG25 protein, and with a 36-kDa protein, distinct from lumican and keratocan. KSPG25-related proteins, not modified with keratan sulfate, were also detected in several connective tissues. Northern blot analysis showed mRNA transcripts of 2.4, 2.5, and 2.6 kilobases in numerous tissues with the 2.4-kilobase transcript enriched in ocular tissues. Ribonuclease protection analysis detected several protected KSPG25 mRNA fragments, suggesting alternate splicing of KSPG25 transcripts. We conclude that the full-length translation product of the gene producing osteoglycin is a corneal keratan sulfate proteoglycan, also present in many non-corneal tissues without keratan sulfate chains. The multiple size protein products of this gene appear to result from in situ proteolytic processing and/or alternative splicing of mRNA. The name mimecan is proposed for this gene and its products. PMID- 9346964 TI - G-Protein-coupled receptors and Fcgamma-receptors mediate activation of Akt/protein kinase B in human phagocytes. AB - Activation of the serine/threonine kinase Akt, also called protein kinase B (PKB), was investigated in human neutrophils. Stimulation of the cells with the chemoattractant fMet-Leu-Phe or the chemokines IL-8 and GROalpha leads to the rapid and transient activation of PKB. Maximum PKB activation correlates with the well documented kinetics of respiratory burst and exocytosis. Wortmannin, a selective inhibitor of phosphoinositide 3-kinases (PI 3-kinases) in neutrophils, abrogates PKB activation. Similarly homo and heterotypic cross-linking of FcgammaIIA and FcgammaIIIB causes a transient activation of PKB that is sensitive to wortmannin treatment. Kinase activity measurements in immunoprecipitates from lysates of the myelocytic GM-1 cells or GM-1/CXCR1 cells, which are transfected with the IL-8 receptor 1, confirmed the transient activation of PKB observed in neutrophils. Stimulation of human monocytes with the CC chemokine RANTES (regulated on activation normal T cell expressed and secreted) also results in the activation of PKB. Preincubation of monocytes and neutrophils with Bordetella pertussis toxin inhibits fMet-Leu-Phe and RANTES-stimulated PKB activation, demonstrating that coupling of the receptors to heterotrimeric Gi-protein is required. The data show, that activation of PKB by Gi-protein-coupled receptors is mediated by PI 3-kinase and suggest that PKB is a constituent of neutrophil activating pathways. PMID- 9346965 TI - The Mr 18,000 subunit of the peripheral-type benzodiazepine receptor exhibits both benzodiazepine and isoquinoline carboxamide binding sites in the absence of the voltage-dependent anion channel or of the adenine nucleotide carrier. AB - The peripheral type benzodiazepine receptor (PBR) binds benzodiazepines such as RO5-4864 and isoquinoline carboxamide derivatives such as PK11195. This receptor includes an Mr 18,000 isoquinoline-binding subunit predominantly located in mitochondrial mem- branes. This protein has been found to copurify with two other mitochondrial proteins, namely the outer membrane voltage-dependent anion channel (VDAC), also known as mitochondrial porin, and the inner membrane adenine nucleotide carrier. In vitro reconstitution experiments suggested that the PBR was a multimeric complex in which the isoquinoline binding site was on the Mr 18,000 subunit, denoted pk18, whereas the benzodiazepine binding site required the association of this subunit with VDAC to be expressed. Untransformed cells of the yeast Saccharomyces cerevisiae are devoid of specific binding sites for isoquinolines and benzodiazepines, whereas yeast cells transformed with a pk18 expressing vector exhibit RO5-4864 and PK11195 binding sites that are pharmacologically identical to those of the PBR. To clarify the role of VDAC and of the adenine nucleotide carrier, if any, in the constitution of the benzodiazepine binding site, yeast host strains were constructed in which the corresponding genes had been knocked out. Mitochondria prepared from pk18 producing cells devoid of either VDAC or adenine nucleotide carrier exhibit both benzodiazepine and isoquinoline carboxamide binding sites with little or no change in the Kd values as compared with the wild-type background. These results rule out the contention that VDAC is indispensable for establishing the benzodiazepine binding site and are in agreement with the hypothesis that the Mr 18,000 subunit carries both the isoquinoline carboxamide and benzodiazepine binding domains. PMID- 9346966 TI - Phosphorylation of Ser465 and Ser467 in the C terminus of Smad2 mediates interaction with Smad4 and is required for transforming growth factor-beta signaling. AB - Members of the Smad family of intracellular signal transducers are essential for transforming growth factor-beta (TGF-beta) to exert its multifunctional effects. After activation of TGF-beta receptors, Smad2 and Smad3 become phosphorylated and form heteromeric complexes with Smad4. Thereafter, these activated Smad complexes translocate to the nucleus, where they may direct transcriptional responses. Here we report that TGF-beta mediates phosphorylation of Smad2 at two serine residues in the C terminus, i.e. Ser465 and Ser467, which are phosphorylated in an obligate order; phosphorylation of Ser465 requires that Ser467 be phosphorylated. Transfection of Smad2 with mutation of Ser465 and/or Ser467 to alanine residues into Mv1Lu cells resulted in dominant-negative inhibition of TGF-beta signaling. These Smad2 mutants were found to stably interact with an activated TGF-beta receptor complex, in contrast to wild-type Smad2, which interacts only transiently. Mutation of Ser465 and Ser467 in Smad2 abrogated complex formation of this mutant with Smad4 and blocked the nuclear accumulation not only of Smad2, but also of Smad4. Thus, heteromeric complex formation of Smad2 with Smad4 is required for nuclear translocation of Smad4. Moreover, peptides from the C terminus of Smad2 containing phosphorylated Ser465 and Ser467 were found to bind Smad4 in vitro, whereas the corresponding unphosphorylated peptides were less effective. Thus, phosphorylated Ser465 and Ser467 in Smad2 may provide a recognition site for interaction with Smad4, and phosphorylation of these sites is a key event in Smad2 activation. PMID- 9346967 TI - Purification, characterization, and localization of an ADP-ribosylactin hydrolase that uses ADP-ribosylated actin from rat brains as a substrate. AB - Mammalian ADP-ribosylation is poorly understood. An ADP-ribosylprotein hydrolase that acted on ADP-ribosylated actin was purified from rat brain. The molecular weight of this enzyme was 62, 000 as determined by SDS-polyacrylamide gel electrophoresis and gel filtration. Enzyme activity with ADP-ribosylated actin as a substrate was inhibited by NAD, ATP, ADP, and ADP-ribose, but not by AMP. Mg2+ increased Vmax. Purified ADP-ribosylactin hydrolase catalyzed the hydrolysis of ADP-ribosylated subunits Gsalpha, Gialpha, and Goalpha and elongation factor-2. After de-ADP-ribosylation by the purified ADP-ribosylactin hydrolase, the proteins were re-ADP-ribosylated by brain mono-ADP-ribosyltransferases and bacterial toxins. The actin that was de-modified by ADP-ribosylactin hydrolase could form actin filaments. Two kinds of monoclonal antibodies against ADP ribosylactin hydrolase were prepared and characterized. In an immunohistochemical study, the plasma membranes and cytoplasmic regions of the nerve cells in the rat brain were immunoreactive. In subcellular fractionation of the brains, most of the ADP-ribosylactin hydrolase activity was found in the cytosol and synaptosome fractions. When the synaptosomes were treated with a hypotonic solution, ADP ribosylactin hydrolase activity was found in the supernatant. Our findings suggest that brain ADP-ribosylactin hydrolase has the important function of polymerizing actin for signal transduction in the cytosol of nerve cells and synaptosomes. PMID- 9346968 TI - Activation mechanism of meprins, members of the astacin metalloendopeptidase family. AB - Meprins are mammalian zinc metalloendopeptidases with protease domains structurally related to astacin, the prototype of the "astacin family" of metalloproteases. Mature, active astacins are produced by proteolytic removal of an activation peptide to generate a new NH2-terminal residue. Structural studies indicate that the NH2-terminal ammonium group inserts into a water-filled cavity adjacent to the active site to form a salt bridge with a Glu residue that is conserved in all astacins. A similar interaction is known to play a crucial role in the activation of trypsin, resulting in the hypothesis that this salt bridge is required for the activation of astacin-like proteases. In this study, we have used the mouse meprin alpha subunit as a model to test this hypothesis of zymogen activation of the astacins. Mutants were generated to vary the NH2-terminal residue of the mature meprin alpha subunit (Asn78) and its putative salt bridge partner (Glu178). In addition, mutants creating NH2-terminal extensions and truncations were expressed in human embryonic kidney 293 cells. The recombinant proteins were activated by limited protease digestion and assayed for enzymatic activity and thermal stability. Point mutations of Asn78 resulted in enzymes with activity comparable to the wild-type enzyme, indicating that the structure of this side chain is not essential for activity. NH2-terminal extension mutants of meprin alpha retained partial activity, with greater decreases against peptide relative to protein substrates. A mutant with a deletion of Asn78 to disrupt salt bridge formation with Glu178 had full activity, indicating that the putative salt bridge with Glu178 is not essential for enzyme activity. However, all changes in meprin alpha subunit NH2-terminal structure were found to decrease the thermal stability of the enzyme. These observations and additional data indicate that the zymogen activation mechanism of meprin and other astacins differs from that of the trypsin family of enzymes, and has some features in common with matrixins. It is proposed that prosequence removal of astacins allows the formation of hydrogen bonds involving the two NH2-terminal residues that are critical for enzyme structure. PMID- 9346969 TI - Function of the R domain in the cystic fibrosis transmembrane conductance regulator chloride channel. AB - For a cystic fibrosis transmembrane conductance regulator (CFTR) channel to enter its open state, serine residues in the R domain must be phosphorylated by cAMP dependent protein kinase, and intracellular ATP must bind to the nucleotide binding folds and subsequently be hydrolyzed. CFTR with its R domain partially removed, DeltaR(708-835)-CFTR, forms a chloride channel that opens independently of protein kinase A phosphorylation, with open probability approximately one third that of the wild type CFTR channel. Deletion of this portion of the R domain from CFTR alters the response of the channel to 5' adenylylimidodiphosphate, pyrophosphate, and vanadate, compounds that prolong burst duration of the wild type CFTR channel but fail to do so in the DeltaR CFTR. In addition, the addition of exogenous unphosphorylated R domain protein, which blocks the wild type CFTR channel, has no effect on the DeltaR-CFTR channel. However, when the exogenous R domain is phosphorylated, significant stimulation of the DeltaR-CFTR channel results; Po increases from 0.10 to 0.22. These data are consistent with a model for CFTR function in which the R domain in the unphosphorylated state interacts with the first nucleotide binding fold to inhibit either binding or hydrolysis of ATP or transduction of the effect to open the pore, but when the R domain is phosphorylated, it undergoes conformational change and interacts at a separate site in the first nucleotide binding fold to stimulate either binding or hydrolysis of ATP or transduction of the effect to open the pore. PMID- 9346970 TI - The kidney androgen-regulated protein promoter confers renal proximal tubule cell specific and highly androgen-responsive expression on the human angiotensinogen gene in transgenic mice. AB - Transgenic mice were generated containing a 1542-base pair fragment of the kidney androgen-regulated protein (KAP) promoter fused to the human angiotensinogen (HAGT) gene with the goal of specifically targeting inducible expression of renin angiotensin system components to the kidney. High level expression of both KAP HAGT and endogenous KAP mRNA was evident in the kidney of male mice from two independent transgenic lines. Renal expression of the transgene in female mice was undetectable under basal conditions but could be strongly induced by administration of testosterone. Testosterone treatment did not cause a transcriptional induction in any other tissues examined. However, an analysis of six androgen target tissues in males revealed that the transgene was expressed in epididymis. No other extra-renal expression of the transgene was detected. In situ hybridization demonstrated that expression of HAGT (and KAP) mRNA in males and testosterone-treated females was restricted to proximal tubule epithelial cells in the renal cortex. Although there was no detectable human angiotensinogen protein in plasma, it was evident in the urine, consistent with a pathway of synthesis in proximal tubule cells and release into the tubular lumen. These results demonstrate that 1542 base pairs of the KAP promoter is sufficient to drive expression of a heterologous reporter gene in a tissue-specific, cell specific, and androgen-regulated fashion in transgenic mice. PMID- 9346971 TI - Mutations in the CYS4 gene provide evidence for regulation of the yeast vacuolar H+-ATPase by oxidation and reduction in vivo. AB - The vma41-1 mutant was identified in a genetic screen designed to identify novel genes required for vacuolar H+-ATPase activity in Saccharomyces cerevisiae. The VMA41 gene was cloned and shown to be allelic to the CYS4 gene. The CYS4 gene encodes the first enzyme in cysteine biosynthesis, and in addition to cysteine auxotrophy, cys4 mutants have much lower levels of intracellular glutathione than wild-type cells. cys4 mutants display the pH-dependent growth phenotypes characteristic of vma mutants and are unable to accumulate quinacrine in the vacuole, indicating loss of vacuolar acidification in vivo. The vacuolar proton translocating ATPases (V-ATPase) is synthesized at normal levels and assembled at the vacuolar membrane in cys4 mutants, but its specific activity is reduced (47% of wild type) and the activity is unstable. Addition of reduced glutathione to the growth medium complements the pH-dependent growth phenotype, partially restores vacuolar acidification, and restores wild type levels of ATPase activity. The CYS4 gene was deleted in a strain in which the catalytic site cysteine residue implicated in oxidative inhibition of the yeast V-ATPase has been mutagenized (Liu, Q., Leng, X.-H., Newman, P., Vasilyeva, E., Kane, P. M., and Forgac, M. (1997) J. Biol. Chem. 272, 11750-11756). This catalytic site point mutation suppresses the effects of the cys4 mutation. The data indicate that the acidification defect of cys4 mutants arises from inactivation of the vacuolar ATPase in the less reducing cytosol resulting from loss of Cys4p activity and provide the first evidence for the modulation of V-ATPase activity by the redox state of the environment in vivo. PMID- 9346972 TI - Biosynthesis of heparin/heparan sulfate. cDNA cloning and expression of D glucuronyl C5-epimerase from bovine lung. AB - Glucuronyl C5-epimerases catalyze the conversion of D-glucuronic acid (GlcUA) to L-iduronic acid (IdceA) units during the biosynthesis of glycosaminoglycans. An epimerase implicated in the generation of heparin/heparan sulfate was previously purified to homogeneity from bovine liver (Campbell, P., Hannesson, H. H., Sandback, D., Roden, L., Lindahl, U., and Li, J.-p. (1994) J. Biol. Chem. 269, 26953-26958). The present report describes the molecular cloning and functional expression of the lung enzyme. The cloned enzyme contains 444 amino acid residues and has a molecular mass of 49,905 Da. N-terminal sequence analysis of the isolated liver enzyme showed this species to be a truncated form lacking a 73 residue N-terminal domain of the deduced amino acid sequence. The coding cDNA insert was cloned into a baculovirus expression vector and expressed in Sf9 insect cells. Cells infected with recombinant epimerase showed a 20-30-fold increase in enzyme activity, measured as release of 3H2O from a polysaccharide substrate containing C5-3H-labeled hexuronic acid units. Furthermore, incubation of the expressed protein with the appropriate (GlcUA-GlcNSO3)n substrate resulted in conversion of approximately 20% of the GlcUA units into IdceA residues. Northern analysis implicated two epimerase transcripts in both bovine lung and liver tissues, a dominant approximately 9-kilobase (kb) mRNA and a minor approximately 5-kb species. Mouse mastocytoma cells showed only the approximately 5-kb transcript. A comparison of the cloned epimerase with the enzymes catalyzing an analogous reaction in alginate biosynthesis revealed no apparent amino acid sequence similarity. PMID- 9346973 TI - Covert video recordings of life-threatening child abuse: lessons for child protection. AB - OBJECTIVE: To describe historic markers and clinical observations of life threatening child abuse as diagnosed using covert video surveillance (CVS). DESIGN: A descriptive, retrospective, partially controlled case study. SETTING: Two hospitals (in London and North Staffordshire, UK) receiving referrals for the investigation of apparent life-threatening events (ALTE), with the availability of CVS. PATIENTS: A total of 39 children (age range at CVS, 2 to 44 months; median, 9 months) in whom hospital CVS was used to investigate suspicions of induced illness. Thirty-six were referred for investigation of ALTE, one with suspected epilepsy, one with failure to thrive, and one with suspected strangulation. A control group consisted of 46 children with recurrent ALTE proven on physiologic recordings to be attributable to a natural medical cause (9 attributable to epileptic seizures, and 37 attributable to respiratory problems). INTERVENTION: Collection of historic details from medical, social service, and police records; interagency collaboration in planning, investigations, and management; development and use of CVS as a clinical tool in the investigation of patients in whom there was suspicion of induced illness. OUTCOME: Confirmation of attempted suffocation or other child abuse from CVS. RESULTS: CVS revealed abuse in 33 of 39 suspected cases, with documentation of intentional suffocation observed in 30 patients. Poisonings (with disinfectant or anticonvulsant), a deliberate fracture, and other emotional and physical abuse were also identified under surveillance. The first ALTE occurred at a median age corrected for the expected date of delivery of 3.6 months in the CVS patients and of 0.3 months in controls. Three CVS patients and 27 of the control children (including 20 at <32 weeks' gestation) were born prematurely. Bleeding from the nose and/or mouth was reported in 11 of the 38 patients with ALTE undergoing CVS but in none of the 46 controls. Four patients who had been subjected to recurrent suffocation before CVS had permanent neurologic deficits and/or required anticonvulsant therapy for epileptic seizures resulting from hypoxic cerebral injury. The 39 patients undergoing CVS had 41 siblings, 12 of whom had previously died suddenly and unexpectedly. Eleven of the deaths had been classified as sudden infant death syndrome but after CVS, four parents admitted to suffocating eight of these siblings. One additional sibling who had died suddenly with rotavirus gastroenteritis was reinvestigated after CVS of her sister revealed poisoning, and death was found to be caused by deliberate salt poisoning. Other signs of abuse were documented in the medical, social, and police records of an additional 15 of the siblings. In the 52 siblings of the 46 controls, 2 had died: one from hypoplastic left heart at 5 days and the other suddenly and unexpectedly (classified as sudden infant death syndrome) at 7 weeks. Twenty-three of the abusive parents were diagnosed by a psychiatrist as having personality disorders. CONCLUSIONS: Induced illness is a severe form of abuse that may cause death or permanent neurologic impairment. It may be accompanied by other severe forms of abuse, may result in behavioral disorders, and may be accompanied by immeasurable suffering. Detection of this abuse requires careful history-taking; thorough examination of the health, social, and police records; and close and focused collaboration between hospital and community child health professionals, child psychiatrists, social workers, and police officers. CVS may help investigate suspicions and ensure that children are protected from additional abuse. When parents have failed to acknowledge that they have deceived health professionals, partnership with them in seeking to protect their children may be neither safe nor effective. PMID- 9346974 TI - A long-term prospective study of varicella vaccine in healthy children. AB - BACKGROUND: Studies in Japan and the United States have shown that varicella vaccine is both safe and efficacious. In 1984, we undertook a 10-year prospective study using a research lot of Oka/Merck varicella vaccine to assess antibody persistence and breakthrough chickenpox rates. In 1987, we began a similar prospective study with lots made in production facilities that ended after 6 years because many children were given a second dose. The purpose of this study is to report humoral antibody persistence and breakthrough chickenpox rates after 6 to 10 years of prospective follow-up. METHODS: One hundred forty-three seronegative children received a research lot (950 plaque-forming units/dose) with 97.9% seroconversion by an assay for fluorescent antibody to membrane antigen (FAMA). One hundred thirty-eight children received production lots (1145 to 3265 plaque-forming units/dose) with 93.5% seroconversion. Yearly chickenpox exposure surveys were completed by phone, and children were seen by a study nurse whenever chickenpox was suspected. A subset in each group had serum collected every 2 years and tested for FAMA antibody. RESULTS: In the research group there have been 25 cases of chickenpox in 137 seroconverters in a period of more than 10 years (yearly rate of 1.7%). In the production lot group there have been 22 cases of chickenpox in 129 seroconverters in a 6-year period (yearly rate of 2.8%). In the research group the median titer rose from 1:16 to 1:64 between 1 and 10 years. In the production group, the median titer did not change between 1, 2, and 4 years. Median antibody titers were compared between the research and production groups at 1, 2, and 4 years and did not differ. The rate of development of modified chickenpox has not increased with time since vaccination, and neither has the case severity. Children with FAMA titers /=64. CONCLUSIONS: 1) Modified chickenpox has occurred in approximately 2% to 3% of vaccinees per year, regardless of the vaccine lot given. 2) FAMA titers have risen between 1 and 10 years in research lot recipients and remained the same in production lot recipients. 3) The likelihood of modified chickenpox developing is inversely related to the 6-week postvaccination FAMA titer. PMID- 9346975 TI - MMR2 immunization at 4 to 5 years and 10 to 12 years of age: a comparison of adverse clinical events after immunization in the Vaccine Safety Datalink project. The Vaccine Safety Datalink Team. AB - BACKGROUND: The Advisory Committee on Immunization Practices recommends a second dose of measles, mumps, and rubella vaccine (MMR2) at age 4 to 5 years of age, whereas the American Academy of Pediatrics suggests MMR2 immunization at age 11 to 12 years of age. Because there is little information on whether the rate of adverse reactions to MMR2 immunization varies among these two age groups, we took advantage of differing immunization policies at two large HMOs to compare the frequency of clinical events after, and possibly related to, MMR2 immunization. METHODS: Information was collected on clinical events plausibly associated to MMR immunization (seizures, pyrexia, malaise/fatigue, nervous/musculoskeletal symptoms, rash, edema, induration/ecchymoses, lymphadenopathy, thrombocytopenia, aseptic meningitis, and joint pain) in two cohorts. At three facilities at Northern California Kaiser (Oakland, CA), 8514 children received MMR2 immunization at age 4 to 6 years of age; at Group Health Cooperative (Seattle, WA) 18 036 children received MMR2 immunization at age 10 to 12 years of age. To account for age-related differences in health care use, within each HMO, clinical events in a 30-day period after immunization were compared with a 30-day period before vaccination. RESULTS: Children 10 to 12 years of age were 50% more likely to have a clinical event after MMR2 immunization than in the period before immunization (odds ratio, 1.45; 95% confidence interval: 1.00,2.10). Children 4 to 6 years of age were less likely to have a visit for an event after immunization compared with the period before immunization (odds ratio, 0.64; 95% confidence interval: 0.40,1.01). CONCLUSIONS: These results suggest that the risk for clinical events after MMR2 immunizations is greater in the 10- to 12-year age group. PMID- 9346976 TI - A safety and immunogenicity comparison of 12 acellular pertussis vaccines and one whole-cell pertussis vaccine given as a fourth dose in 15- to 20-month-old children. AB - OBJECTIVE: To compare the safety and immunogenicity of 12 different acellular pertussis vaccines combined with diphtheria and tetanus toxoids (DTaP) with one licensed diphtheria, tetanus, and whole-cell pertussis vaccine (DTwP) as a fourth dose booster in children who had previously received DTaP or DTwP primary vaccinations. METHODS: Healthy 15- to 20-month-old children were enrolled at six National Institutes of Health Vaccine Treatment and Evaluation Units. All had been randomly assigned to receive three primary doses of DTaP or DTwP at 2, 4, and 6 months of age as part of an earlier National Institutes of Health multicenter trial of DTaP vaccines in the same Vaccine Treatment and Evaluation Units. Parents recorded the occurrence and magnitude of fever; irritability; and injection site redness, swelling, and pain for 3 days after vaccination. Sera obtained before and 1 month after the booster vaccination were analyzed for antibody to pertussis toxin (PT), filamentous hemagglutinin (FHA), fimbriae (FIM), and pertactin (PRN). Diphtheria and tetanus toxoid as well as PT neutralizing (Chinese hamster ovary cell) and whole-cell agglutinating antibodies were measured on a subset of sera. RESULTS: A total of 1293 children contributed fourth-dose reaction data. Reactions were less frequent after DTaP than after DTwP. For children vaccinated with a fourth dose of DTaP, which was the same DTaP as received in the primary series, fever and injection site redness, swelling, and pain increased in prevalence compared with the third dose in the primary series. For children receiving DTaP as a fourth dose, injection site redness and swelling occurred more frequently in DTaP-primed than in DTwP-primed children. Variation in the occurrence of reactions among DTaP vaccines was observed. A total of 1160 paired pre- and postvaccination sera were available for analysis. Serum antibody concentrations before boosting were lower than those obtained 1 month after the primary immunization. After the fourth dose, significant increases in antibodies directed against the included antigens were observed for all vaccines; postbooster vaccination antibody titers differed significantly among the DTaP vaccines. For children primed and boosted with the same DTaP, antibody levels were not directly related to the quantity of antigen included for PT, FHA, and FIM; for PRN, there was a closer relationship. Some DTaP vaccines given as fourth-dose boosters elicited antibody to PRN or FIM in some vaccinees, although the DTaP vaccines were not reported to contain these antigens; these responses were observed more frequently in DTwP-primed children. Agglutinin antibody rises were observed in all groups immunized with four doses of a DTaP vaccine containing FHA or PRN, regardless of whether the vaccine included FIM. Diphtheria and tetanus antibody levels exceeded the presumed protective concentration (0.1 IU/mL for diphtheria and 0.01 IU/mL for tetanus) after the fourth dose for all vaccinees. CONCLUSION: Although differences were observed in reaction rates among the DTaP vaccines given as a fourth dose, the DTaP vaccines were, in general, associated with fewer adverse events than a US-licensed DTwP. For DTaP vaccines, fever; irritability; and injection site pain, redness, and swelling occurred more frequently after the fourth dose than after the third dose of the same vaccine in the primary series. No DTaP was consistently most or least reactogenic or immunogenic. Although serologic correlates of pertussis immunity are not defined, it is clear that most DTaP vaccines can stimulate comparable or higher serum antibody responses than DTwP for those antigens contained in the vaccine. PMID- 9346977 TI - Interleukin-6 in the cerebrospinal fluid after perinatal asphyxia is related to early and late neurological manifestations. AB - OBJECTIVES: To investigate if the concentration of interleukin-6 (IL-6) in the cerebrospinal fluid (CSF) is affected by perinatal asphyxia, and to examine the relation of IL-6 levels in the CSF to the severity of hypoxic-ischemic encephalopathy (HIE), to brain damage, and to the neurological outcome. METHODS: Asphyxiated term neonates were included. Cerebrospinal fluid IL-6 was measured by a sensitive enzyme-linked immunosorbent assay. RESULTS: Twenty neonates were studied: 3 had no HIE, 5 had stage 1, 6 had stage 2, and 6 had stage 3. CSF IL-6 levels (8 to 90 hours of life) were higher in neonates with HIE stage 3 (range, 65 to 2250 pg/mL) when compared with neonates with HIE stage 0 to 2 (<2 pg/mL in 12 neonates, 10 pg/mL in 1). According to neuroimaging techniques and/or pathological examination, 14 neonates were normal, and 5 showed signs of brain damage; 1 was not classified. CSF IL-6 levels were significantly higher in neonates with signs of brain damage. Finally, 5 neonates had adverse outcomes (4 died, 1 had cerebral palsy), 2 had mild motor impairment, and 13 had normal outcomes. CSF IL-6 levels were higher in neonates with adverse outcomes (range, 65 to 2250 pg/mL) compared with neonates with favorable outcomes. CONCLUSION: The magnitude of IL-6 response in the CSF after perinatal asphyxia is related to the severity of neonatal HIE, to brain damage, and to the neurological outcome. Our results suggest that IL-6 might play a role in neonatal hypoxic-ischemic brain damage. PMID- 9346978 TI - Margin of safety for discharge after apnea in preterm infants. AB - OBJECTIVE: Most neonatologists include an apnea-free period in the criteria for the discharge of preterm infants. However, the length of time one should wait after the cessation of apnea before sending an infant home without a monitor is debated. We undertook this study in an attempt to define a minimal and safe observation period between the time of the last apnea episode and discharge. METHODS: We reasoned that in infants with idiopathic apnea of prematurity, the intervals between days on which apnea occurs gradually increase until some point at which clinically significant apnea ceases. Therefore, knowledge about the intervals between days on which apnea occurred just before the last apnea would provide a reasonable estimate of the minimal safe observation interval between the last apnea and discharge. We reviewed the charts of 266 infants born in 1993 and 1994 at /=25 years of age) had the most favorable outcomes, and those of teenage mothers (<20 years of age) had the least favorable outcomes; 22% of daughters and 6% of sons of the oldest mothers versus 38% and 18%, respectively, of the youngest mothers became teenage parents. CONCLUSION: The mother's age at delivery is an independent determinant of the child's adult status. PMID- 9346980 TI - Regulation of growth of 7- to 36-month-old children by energy and fat intake in the prospective, randomized STRIP baby trial. AB - OBJECTIVE: To study the fat and energy intakes of children between 7 and 36 months of age with different growth patterns. METHODS: In the Special Turku coronary Risk factor Intervention Project for Babies, children were randomized to intervention (n = 540) and control groups (n = 522) at age 7 months. The intervention was aimed at replacing part of the saturated fat intake with monounsaturated and polyunsaturated fat to reduce children's exposure to high serum cholesterol values. The control children consumed a free diet. Children followed for >2 years (n = 848) were included in the analysis. Five groups of children representing different extreme growth patterns during the first 3 years of life were formed, and their energy and fat intakes were analyzed. Relative weight was defined as deviation of weight in percentages from the mean weight of healthy children of same height and sex, and relative height as deviation of height in SD units from the mean height of healthy children of same age and sex. RESULTS: Relative fat intakes (as percent of energy intake) were similar in children showing highly different height gain patterns. The thin (mean relative weight /= 95%) and the obese (mean relative weight >/= 95%) were highest, but weight-based energy intake of the tall (at 2 years, 82 [13] kcal/kg) and the obese (79 [17] kcal/kg) were lower than that of children with normal growth (89 [16] kcal/kg). The thin children consumed relatively more energy than the children with normal growth (at 2 years, 94 [13] kcal/kg and 89 [16] kcal/kg, respectively). Parental height and body mass index and the child's absolute and relative energy intakes predicted the best children's growth patterns. Children with consistently low fat intake grew equally to the children with higher fat intake. CONCLUSIONS: Moderate supervised restriction of fat intake to values 25 to 30 E% is compatible with normal growth. PMID- 9346981 TI - Longitudinal neurological follow-up of a group of HIV-seropositive and HIV seronegative hemophiliacs: results from the hemophilia growth and development study. AB - BACKGROUND: Boys and young men with hemophilia treated with factor infusions before 1985 had a substantial risk of acquiring the human immunodeficiency virus (HIV) and the acquired immunodeficiency syndrome. This study was designed to assess the effects of HIV and hemophilia per se on neurological function in a large cohort of subjects with hemophilia, and to investigate the relationships between neurological disease and death during follow-up. METHODS: Three hundred thirty-three boys and young men (207 HIV seropositive and 126 HIV seronegative) were evaluated longitudinally in a multicenter, multidisciplinary study. Neurological history and examination were conducted at baseline and annually for 4 years. The relationship between neurological variables, HIV serostatus, CD4+ cell counts, and vital status at the conclusion of the study was examined using logistic regression models. RESULTS: The risks of nonhemophilia-associated muscle atrophy, behavior change, and gait disturbance increased with time in immune compromised HIV-seropositive subjects compared with HIV seronegative or immunologically stable HIV-seropositive subjects. The risk of behavior change in immune compromised HIV-seropositive hemophiliacs, for example, rose to 60% by year 4 versus 10% to 17% for the other study groups. Forty-five subjects (13.5%), all of whom were HIV seropositive, died by year 4. Subjects who died had had increased risks of hyperreflexia, nonhemophilia-associated muscle atrophy, and behavior change. CONCLUSIONS: These results indicate that immune compromised, HIV seropositive hemophiliacs have high rates of neurological abnormalities over time and that neurological abnormalities were common among subjects who later died. By contrast, immunologically stable HIV-seropositive subjects did not differ from the HIV-seronegative participants. Hemophilia per se was associated with progressive abnormalities of gait, coordination, and motor function. PMID- 9346983 TI - Hypospadias trends in two US surveillance systems. AB - OBJECTIVE: Hypospadias is a common congenital anomaly, the cause of which is unknown. Unexplained increases in the rates of hypospadias occurred in five European countries in the 1970s and 1980s. We examined data from two birth defects surveillance systems in the United States for evidence of similar trends. METHODOLOGY: The Metropolitan Atlanta Congenital Defects Program (MACDP) provided birth prevalence rates from 1968 to 1993. The nationwide Birth Defects Monitoring Program (BDMP) provided rates from 1970 to 1993. MACDP data are population-based and could be categorized by the severity of the hypospadias. BDMP data allowed analysis of rate trends for the four census regions of the United States. RESULTS: Data from both surveillance systems showed an approximate doubling of hypospadias rates in the 1970s and 1980s. MACDP data showed that the rate of severe cases increased while the ratio of mild to severe cases decreased. BDMP data showed that hypospadias rates increased markedly in all four regions of the United States. CONCLUSIONS: The observed increases are unlikely to be attributable to increased sensitivity of the surveillance systems or the identification of more mild cases by physicians over time, because either trend would have increased rather than decreased the ratio of mild to severe cases. If real, these trends represent the largest number of cases and the first report of an increase in hypospadias rates outside of Europe. Additional investigation of a possible increase in hypospadias rates is warranted. PMID- 9346982 TI - New non-cocaine-containing topical anesthetics compared with tetracaine adrenaline-cocaine during repair of lacerations. AB - OBJECTIVE: To compare the effectiveness of three new topical anesthetics that do not contain cocaine (prilocaine-phenylephrine, tetracaine-phenylephrine [tetraphen], and tetracaine-lidocaine-phenylephrine) to that of tetracaine adrenaline-cocaine (TAC) during laceration repair in children. DESIGN: Prospective, randomized, double-blind clinical trial. SETTING: The emergency department of an urban children's hospital. PARTICIPANTS: Children 1 year of age or older with a laceration /= 5 years of age using a visual analogue scale (VAS). Suture technicians, research assistants, and parents also scored pain using a seven-point Likert scale. In addition, suture technicians completed an anesthetic effectiveness scale. RESULTS: There was consistently no difference demonstrated between the effectiveness of tetraphen and that of TAC for each outcome measure of each observer group. A statistically significant difference was seen among anesthetics when comparing VAS and Likert scale scores of suture technicians and Likert scale scores of research assistants. Based on post hoc analyses, these statistically significant differences were between TAC and prilocaine-phenylephrine (suture technician VAS and Likert scale) and between TAC and tetracaine-lidocaine-phenyl-ephrine (suture technician Likert scale), but not between TAC and tetraphen. When power analyses were performed using alpha = 0.05 and beta = 0.20, it was possible to detect a difference of 1.2 VAS units for each of the observer groups. Based on anesthetic effectiveness scale scores, the three new topical preparations collectively performed significantly better on the face and scalp than on the extremities (relative risk = 1.83; 95% confidence interval 1.20 < relative risk < 2.79). CONCLUSION: This study demonstrated the effectiveness and safety of three new non cocaine-containing topical anesthetics. Consistently, there was no statistical difference demonstrated between the effectiveness of tetraphen and that of TAC for each outcome measure of each observer group. Tetraphen offers an effective alternative to TAC during laceration repair in children. PMID- 9346984 TI - Risk factors for sudden infant death syndrome following the prevention campaign in New Zealand: a prospective study. AB - OBJECTIVES: To identify the risk factors for sudden infant death syndrome (SIDS) following a national campaign to prevent SIDS. METHODS: For 2 years (October 1, 1991 through September 30, 1993) data were collected by community child health nurses on all infants born in New Zealand at initial contact and at 2 months. RESULTS: There were 232 SIDS cases in the postneonatal age group (2.0/1000 live births) and these were compared with 1200 randomly selected control subjects. Information was available for 127 cases (54.7%) and 922 (76.8%) of controls. The previously identified modifiable risk factors were examined. The prevalence of prone sleeping position of the infant was very low (0.7% at initial contact and 3. 0% at 2 months), but was still associated with an increased risk of SIDS. In addition, the side sleeping position was also found to have an increased risk of SIDS compared with the supine sleeping position (at 2 months: adjusted odds ratio (OR) = 6.57; 95% confidence interval (CI) = 1.71, 25.23). Maternal smoking was found to be the major risk factor for SIDS. Bed sharing was also associated with an increased risk of SIDS. There was an interaction between maternal smoking and bed sharing on the risk of SIDS. Compared with infants not exposed to either bed sharing or maternal smoking, the adjusted OR for infants of mothers who smoked was 5.01 (95% CI = 2.01, 12.46) for bed sharing at the initial contact and 5.02 (95% CI = 1.05, 24. 05) for bed sharing at 2 months. In this study breastfeeding was not associated with a statistically significant reduction in the risk of SIDS. The other risk factors for SIDS identified were: unmarried mother, leaving school at a younger age, young mother, greater number of previous pregnancies, late attendance for antenatal care, smoking in pregnancy, male infant, Maori ethnicity, low birth weight, and shorter gestation. CONCLUSIONS: After adjustment for potential confounders, prone and side sleeping positions, maternal smoking, and the joint exposure to bed sharing and maternal smoking were associated with statistically significant increased risk of SIDS. A change from the side to the supine sleeping position could result in a substantial reduction in SIDS. Maternal smoking is common in New Zealand and with the reduction in the prevalence of prone sleeping position is now the major risk factor in this country. However, smoking behavior has been difficult to change. Bed sharing is also a major factor but appears only to be a risk to infants of mothers who smoke. Addressing bed sharing among mothers who smoke could reduce SIDS by at least one third. Breastfeeding did not appear to offer a statistically significant reduction in SIDS risk after adjustment of potential confounders, but as breastfeeding rates are comparatively good in New Zealand, this result should be interpreted with caution as the power of this study to detect a benefit is small. PMID- 9346985 TI - Infant arousals during mother-infant bed sharing: implications for infant sleep and sudden infant death syndrome research. AB - OBJECTIVE: Normative values for infant sleep architecture have been established exclusively in the solitary sleeping environment. However, most of the world's cultures practice some form of parent-infant cosleeping. In addition, no previous polysomnographic studies in infants examined the frequency of electroencephalogram (EEG) arousals. This is the first study to assess (a) EEG arousals in infants and their relationship to sleep stages; (b) the impact on arousals of mother-infant bed sharing; and (c) the temporal overlap of infant with maternal arousals during bed sharing. METHODOLOGY: Three nights of polysomnography were performed in 35 breastfeeding mother-infant pairs when the infants were 11 to 15 weeks old. An adaptation night was followed by one bed sharing night and one solitary sleeping night. Twenty infants had been bed sharing since birth and 15 were routine solitary sleepers. Both epochal awakenings (EWs), based on 30-second epoch scoring of sleep-wake stages, and more transient arousals (TAs) >/=3 seconds were quantified. RESULTS: Stage 3-4 sleep was associated with a striking paucity of EWs and TAs compared with stages 1-2 or rapid eye movement sleep. Bed sharing facilitated EWs and TAs selectively during stage 3-4 sleep. EWs from stage 3-4 sleep were more frequent on the bed sharing night than on the solitary night in both infant groups. Routinely bed sharing infants also exhibited more frequent TAs in stage 3-4 than the routine solitary sleepers in both conditions. In both groups, the number of infant arousals (EWs + TAs) that overlapped the mother's was doubled during bed sharing, with infant arousals leading most often. CONCLUSIONS: Mother-infant bed sharing promotes infant arousals. Together with a previous report that bed sharing reduces stage 3 4 sleep, this suggests that normative values for infant sleep must be interpreted within the context of the sleeping environment in which they were established. Given that arousability is diminished in stage 3-4, we speculate that, under otherwise safe conditions, the observed changes in stage 3-4 sleep and arousals associated with bed sharing might be protective to infants at risk for SIDS because of a hypothesized arousal deficit. The responsivity of the mother to infant arousals during bed sharing might also be protective. PMID- 9346986 TI - Illnesses and absence due to illness among children attending child care facilities in Seattle-King County, Washington. AB - OBJECTIVES: Although much of the economic impact of child care-associated illness in the United States is due to parents' time lost from work, there are no data on the incidence of absence due to illness among children in various types of out-of home child care settings in the United States. The goals of this study were to compare the incidence of illness and absence due to illness among children attending child care homes (CCHs) and child care centers (CCCs). METHODS: From July 1992 through June 1993, child care providers from 91 CCHs and 41 CCCs in Seattle-King County, Washington, provided information on absenteeism and illness for 96 792 child-weeks of observation. RESULTS: The age-adjusted incidence of provider-reported illness episodes among children in CCHs (10.4 episodes per 100 child-weeks) was greater than that among children in CCCs (6.7 episodes per 100 child-weeks). The incidence density ratio of illness among children <1 year of age in comparison to those >/=5 years of age in CCCs (4.5) was greater than that among similar groups in CCHs (2.3). The age-adjusted incidence of absence due to illness among children in CCHs (5.1 days per 100 child-weeks) was less than that among children in CCCs (8.9 days per 100 child-weeks). CONCLUSIONS: Results comparing the incidence of illness between children in various types of child care settings may be influenced by information sources. The incidence of illness among children in CCHs may be greater than that among children in CCCs. The increased incidence of absence due to illness among children in CCCs compared with that among children in CCHs probably reflects differences in exclusion and attendance policies and practices between these two types of settings. PMID- 9346987 TI - Decrease in birth weight in relation to maternal bone-lead burden. AB - OBJECTIVES: Birth weight predicts infant survival, growth, and development. Previous research suggests that low levels of fetal lead exposure, as estimated by umbilical cord blood-lead levels at birth, may have an adverse effect on birth weight. This report examines the relationship of lead levels in cord blood and maternal bone to birth weight. METHODS: Umbilical cord and maternal venous blood samples and anthropometric and sociodemographic data were obtained at delivery and 1-month postpartum. Blood-lead levels were analyzed by atomic absorption spectrophotometry. Maternal tibia and patella lead levels were determined at 1 month postpartum with use of a spot-source 109Cd K-X-ray fluorescence instrument. The relationship between birth weight and lead burden was evaluated by multiple regression with control of known determinants of size at birth. RESULTS: Data on all variables of interest were obtained for 272 mother-infant pairs. After adjustment for other determinants of birth weight, tibia lead was the only lead biomarker clearly related to birth weight. The decline in birth weight associated to increments in tibia lead was nonlinear and accelerated at the highest tibia lead quartile. In the upper quartile, neonates were on average, 156 grams lighter than those in the lowest quartile. Other significant birth weight predictors included maternal nutritional status, parity, education, gestational age, and smoking during pregnancy. CONCLUSIONS: Our results indicate that bone-lead burden is inversely related to birth weight. Taken together with other research indicating that lead can mobilize from bone into plasma without detectable changes in whole blood lead, these findings suggest that bone lead might be a better biomarker than blood lead. Because lead remains in bone for years to decades, mobilization of bone lead during pregnancy may pose a significant fetal exposure with health consequences, long after maternal external lead exposure has declined. PMID- 9346988 TI - The effect of changes in dietary fat on the food group and nutrient intake of 4- to 10-year-old children. AB - OBJECTIVE: To determine how young children changed their overall diet when they changed their fat intake after 3 months of participating in a nutrition education demonstration study designed to lower low-density lipoprotein cholesterol and cardiovascular risk. METHODS: Three 24-hour dietary recalls were collected from 303 4- to 10-year-old children at baseline and 3 months later. At both times, mean number of servings from food groups, grams of fat contributed from food groups, and intake of calories and nutrients were calculated and compared among quartiles of children formed according to change in their percent of calories from total fat after 3 months. RESULTS: Children who reduced their percent of calories from total fat most (ie, by an average of 8.5%) after 3 months consumed fewer servings from meats, eggs, dairy, fats/oils, and breads but tended to increase their number of servings from lower-fat foods within those food groups, particularly from dairy foods. These children also increased their mean intake of fruits, vegetables, and desserts, and maintained average intakes of all nutrients (except vitamin D) in excess of two thirds of the respective recommended dietary allowance. CONCLUSIONS: Young children who reduced their percent of calories from total fat in accordance with the current National Cholesterol Education Program recommendations accomplished this by reducing their overall intake of higher-fat foods, replacing higher-fat foods with lower-fat foods within several food groups, particularly within the dairy group (eg, drinking skim milk instead of whole milk) and by consuming more servings of fruits, vegetables, and very-low fat desserts. These behaviors did not compromise their mean calorie or nutrient intakes, showing that it is possible for young children to lower their fat intake safely to reduce their risk of future heart disease. PMID- 9346989 TI - War experiences and distress symptoms of Bosnian children. AB - OBJECTIVE: The war in Bosnia has had a tremendous impact on civilians. Little is known about the impact of modern warfare on children. This survey documents the nature and frequency of war-related experiences among Bosnian children and describes their manifestations of selected psychological sequelae. METHODS: A cross-sectional survey of 364 internally displaced 6- to 12-year-old children and their parents living in central Bosnian collectives was conducted during the war. Parents were surveyed for their children's war experiences; the children were surveyed for war-related distress symptoms. RESULTS: The children were exposed to virtually all of the surveyed war-related experiences. The majority had faced separations from family, bereavement, close contact with war and combat, and extreme deprivation. The prevalence and severity of experiences were not significantly related to a child's gender, wealth, or age, but were related to their region of residence, with children from the region of Sarajevo having the highest prevalence of experiences. Almost 94% of the children met Diagnostic and Statistical Manual of Mental Disorders, 4th ed, criteria for posttraumatic stress disorder. Significant life activity affecting sadness and anxiety were reported by 90.6% and 95.5% of the children, respectively. High levels of other symptoms surveyed were also found. Children with greater symptoms had witnessed the death, injury, or torture of a member of their nuclear family, were older, and came from a large city. CONCLUSIONS: The war-related experiences of the children studied were both varied and severe, and were associated with a variety of psychological sequelae. This experience underscores the vulnerability of civilians in areas of conflict and the need to address the effects of war on the mental health of children. PMID- 9346990 TI - The effect of recombinant human growth hormone in children with X-linked hypophosphatemia. AB - BACKGROUND: X-linked hypophosphatemia (XLH) is characterized clinically by rickets and growth retardation. Conventional treatment of XLH with oral phosphate and vitamin D fails to normalize linear growth. Objective. To determine the benefit and the potential side effects of recombinant human growth hormone (rhGH) therapy in patients with XLH. DESIGN AND METHODS: A randomized, double-blind, crossover study was performed throughout a 24-month period in five children with XLH, each patient serving as his own control. The effect of 12 months of rhGH therapy on height, mineral metabolism, glucose and lipid metabolism, hemoglobin, thyroid and parathyroid function, serum 1,25-(OH)2 vitamin D, osteocalcin, growth hormone, urinary calcium, phosphate, nephrocalcinosis, renal function, and bone density was compared with the effects of 12 months of placebo administration on the same parameters. RESULTS: The average age (mean +/- SEM) of the patients at the start of the study was 5.6 +/- 1.4 years. Growth hormone therapy improved the height standard deviation score (z-score) from a baseline of -2.66 +/- 0.21 to 2.02 +/- 0.25 and to -1.46 +/- 0.28, after 3 and 12 months, respectively. At the start of the control period the height z-score was -2.27 +/- 0.30 compared with 2.22 +/- 0.16 after 12 months of placebo administration. The growth velocity standard deviation score was -1. 90 +/- 0.40 during the 12 months of placebo administration and +4.04 +/- 1.50 during the 12 months of rhGH therapy. An increase in serum phosphate from 0.88 +/- 0.07 mmol/L to 1.17 +/- 0.14 mmol/L and tubular maximum for phosphate reabsorption (TmP/GFR) from 2.12 +/- 0. 15 to 3.41 +/- 0.25 mg/dL, was observed after 3 months of rhGH therapy. However, both serum phosphate and TmP/GFR were unchanged from baseline after 6, 9, and 12 months of rhGH therapy. Neither serum phosphate nor TmP/GFR changed from baseline during the placebo administration. Insulin-like growth factor 1 (IGF-1) increased from 114 +/- 25 to 354 +/- 51 ng/mL after 12 months of rhGH therapy. Despite the increase in IGF-1 after rhGH therapy, the value did not exceed normal serum concentration. IGF-1 did not change from baseline after 12 months of placebo administration. Neither therapy with rhGH nor with placebo had an effect on glucose and lipid metabolism, hemoglobin, thyroid and parathyroid function, serum 1, 25-(OH)2 vitamin D, alkaline phosphatase, osteocalcin, urinary calcium excretion, the grade of nephrocalcinosis, glomerular filtration rate, or urinary albumin excretion. Twelve months of rhGH therapy increased bone mass and width but not density. Twelve months of placebo administration had no effect on bone mass, width, or density. CONCLUSION: Patients with XLH have an improvement in linear growth and a transient increase in serum phosphate attributable to a transient decrease in urinary phosphate excretion when treated with rhGH. PMID- 9346991 TI - Unimaginable images: seeing is believing. PMID- 9346992 TI - Harmony on the second dose of measles vaccine. PMID- 9346993 TI - Introduction to neonatal systematic reviews. PMID- 9346994 TI - Large intergenerational variation in age of onset in two young patients with Huntington's disease presenting as dyskinesia. PMID- 9346995 TI - Perinatal substance abuse: the impact of reporting infants to child protective services. AB - OBJECTIVE: The purposes of this study were to follow the judicial placement of newborns with positive toxicology screening results and to determine how long such infants remained in foster care, separated from their mothers or other relatives, and the length of court dependency. We also determined the mothers' compliance with court orders, the availability and use of rehabilitative services, factors used by the court to determine the final disposition, and the eventual placement of the infants. METHODS: The cohort sample consisted of all infants from San Mateo County (CA) born at Stanford University Hospital during a 2-year period whose urine tests in the well-baby nursery were positive for illicit substances. Fifty-three newborns were identified, and their medical records and court documents were matched and reviewed from birth until termination of judicial review (or 5 years). Data were summarized and analyzed by logistic regressions to identify predictors of specific outcomes. RESULTS: All 53 infants had normal physical examinations and uneventful hospital courses. Their ethnic distribution, with 68% being African-American and 7% being Hispanic, differed from the rest of the nursery population, which was predominantly Hispanic. Twenty-six (46%) of the 53 infants were returned to their mothers within 1 week of birth; 39 (76%) of the infants were reunited with some relative within the first month of life. At 12 months of age, 10 infants (19%) remained in foster care; however, none remained in foster care beyond 18 months. The length of time infants were dependents of the court ranged from 1 month to >5 years; 70% of the cases were "closed" between 6 and 30 months of life. Nine (17%) were dependents of the court for >36 months. Final placement of the infants was 35 (66%) reunited with at least one parent, 9 (17%) in long-term guardianship relationships with other relatives, and 9 (17%) adopted. All of the mothers were ordered to complete a drug rehabilitation program; 24 mothers (44%) fully complied and had repeatedly drug-free urine tests; 2 others (4%) had drug-free urine tests after incomplete participation in drug rehabilitation. Twenty-two (42%) of the mothers never complied with drug rehabilitation. Subsequent drug use was evident in less than half of the mothers during the period of study. Only one mother was reported for child abuse. Characteristics that most strongly predicted failure in family reunification were a history of failed drug rehabilitation, previous involvement of Child Protective Services, or previous removal of a child because of substance abuse. CONCLUSION: Identifying and reporting newborns exposed to maternal substance abuse during pregnancy can be associated with beneficial changes in the environment of the infants and successful rehabilitation of many mothers. The use of judicial supervision, rehabilitative and supportive services, and long-term involvement of social services without criminal prosecution are key to successful outcome. This study supports the policy and recommendation of the American Academy of Pediatrics and should lessen health professionals' concerns about negative effects of reporting these patients to Child Protective Services. PMID- 9346996 TI - Acquisition of Pseudomonas aeruginosa in children with cystic fibrosis. AB - OBJECTIVE: This study was pursued as an extension of a randomized clinical investigation of neonatal screening for cystic fibrosis (CF). The project included assessment of respiratory secretion cultures for pathogens associated with CF. The objective was to determine whether patients diagnosed through neonatal screening and treated in early infancy were more likely to become colonized with Pseudomonas aeruginosa compared with those identified by standard diagnostic methods. METHODOLOGY: The design involved prospective cultures of respiratory secretions obtained generally by oropharyngeal swabs at least every 6 months and more often if clinically indicated. Patients were managed with a standardized evaluation and treatment protocol at the two Wisconsin certified CF centers; however, there were community and environmental variations associated with the follow-up period as described below. RESULTS: Overall, there were no differences in acquisition of respiratory pathogens between the screened and the control (standard diagnosis) groups. Evaluation of the data between and within the two centers, however, revealed significant differences with earlier acquisition of P aeruginosa in the center with the following distinguishing characteristics: urban location; following patients with the standard US approach in which newly diagnosed, young children were interspersed with older CF patients; and where there were more opportunities for social interactions with other CF patients. The differences were confined to the screened group followed in the urban center in which the median pseudomonas-free survival period was 52 weeks contrasted with 289 weeks in the other center. In addition, assessment of data for the entire CF populations followed at the two centers revealed that the urban center showed a significantly higher prevalence of P aeruginosa colonization in patients between the ages of 3 and 9 years. CONCLUSIONS: These results present questions and generate hypotheses on risk factors for acquisition of P aeruginosa in CF and suggest that clinic exposures and/or social interactions may predispose such patients to pseudomonas infections. PMID- 9346997 TI - Safety and effectiveness of homemade and reconstituted packet cereal-based oral rehydration solutions: a randomized clinical trial. AB - OBJECTIVES: Parents may be deterred from obtaining commercial oral rehydration solutions (ORS) for their young children with acute diarrheal disease because of its availability and/or cost, especially if they are poor. We conducted a randomized clinical trial to determine 1) whether low-income parents could safely mix and administer cereal-based ORS (CBORS) both from ingredients commonly found in the home and from a premixed packet; 2) whether these CBORS were as effective in maintaining hydration as commercial glucose-based ORS; and 3) whether CBORS were more effective in reducing severity and duration of illness. METHODS: Children 4 to 36 months of age discharged from emergency departments and health centers with acute diarrheal disease were randomized to receive either homemade CBORS, reconstituted packet CBORS, or Pedialyte. A study nurse saw the child at home each day until the illness resolved, and obtained capillary blood for serum sodium at enrollment and at 24 to 48 hours; a sample of CBORS for sodium concentration; stool for pathogen analysis; and daily fluid intake, stool frequency, and weight. RESULTS: A total of 232 children were enrolled, of whom 203 (88%) completed the study. Two parents (3%) in the homemade CBORS group and one parent (1%) in the packet CBORS group made mixing errors resulting in a high sodium concentration (>100 mEq/L); their children refused the solution and had normal serum sodium values. Mean CBORS sodium concentration for the remainder of the homemade CBORS group was 60 +/- 10 mEq/L, and for the packet CBORS group, 54 +/- 13. Eighteen children (11%) had abnormal serum sodium values at presentation, which returned to normal in all groups in most cases. Three children (4.5%) in the homemade CBORS group, 4 (6%) in the packet CBORS group, and 1 child (1.4%) in the Pedialyte group failed therapy. Children refused to take homemade CBORS and packet CBORS (43% and 32%, respectively) more often than Pedialyte (9%), and those in the CBORS groups tended to take less ORS and total fluids. There were no significant differences among the three groups in incidence of daily vomiting or stooling, duration of diarrhea, or weight gain. CONCLUSIONS: CBORS do not offer a clinically significant advantage over glucose-based ORS. Homemade CBORS represent a treatment option in carefully selected cases, but it is not the safest alternative for regular clinical use. PMID- 9346998 TI - Vitamin-mineral supplement use among preschool children in the United States. AB - OBJECTIVE: To estimate the prevalence of recent supplement use in a national sample of preschool children and to examine the relationship of maternal and child characteristics, past maternal supplement use practices, familial, health services, and child health factors associated with supplement use. METHODS: We used data on 8285 preschool children whose mothers were interviewed for the 1991 Longitudinal Follow-up to the 1988 National Maternal and Infant Health Survey. Data collection was conducted either by telephone or personal interview. The sample is representative of the estimated 3. 8 million US born children in 1988 and alive in 1991. The outcome measures are whether the child was given any vitamin and mineral supplements at least 3 days a week in the 30 days before the interview and the type of supplement received. Statistical techniques included bivariate and weighted multiple logistic regression analysis. RESULTS: More than half of all US 3-year-olds (54.4%) were given some vitamin and mineral supplement. The most common supplements consumed were multivitamin-mineral with iron (59% of supplement users) and multivitamin-mineral without iron (26.4%). Children who received any supplements tended to have mothers who are non-Hispanic White, older, more educated, married, insured, receiving care from a private health care provider, have greater household income, and took supplements during pregnancy. Child health characteristics associated with supplement use included first birth order and having eating problems or poor appetites. CONCLUSIONS: More than half of US preschool children used vitamin and mineral supplements. The sociodemographic and health predictors identified for supplement use suggest that groups at risk for nonuse are likely the same groups whose circumstances may predispose a need for supplementation. PMID- 9346999 TI - Pulmonary administration of gentamicin during liquid ventilation in a newborn lamb lung injury model. AB - OBJECTIVES: Newborns with pulmonary infection frequently present with acute lung injury leading to ventilation/perfusion abnormalities in which intravenous delivery of antibiotics to the lung can be suboptimal. Tidal liquid ventilation (TLV) has been shown to be an effective means for delivering drugs directly to the pulmonary system. The objective of this study was to compare, with lung injury, antibiotic delivery achieved by conventional techniques (gas ventilation and intravenous gentamicin) with that using pulmonary administration of drug (PAD) during TLV. METHODS: Twelve newborn lambs with an acid lung injury were randomized to receive gentamicin either intravenously during gas ventilation or via PAD during TLV using LiquiVent (Alliance Pharmaceutical Corporation, San Diego, CA, and Hoechst-Marion Roussel, Bridgewater, NJ) perfluorochemical. Gentamicin (5 mg/kg) was administered over 1 minute, and serum levels were obtained at 15-minute intervals. Arterial blood gases and pulmonary mechanics were measured. Ventilation efficiency index and arterial/alveolar oxygen ratio were calculated. Lung-tissue gentamicin levels were measured 4 hours after administration and corrected to dry weight. RESULTS: Serum gentamicin levels were similar in both groups. Lung gentamicin levels (micrograms/g) were significantly higher for TLV. Also, TLV resulted in significantly more of the total delivered dose in the lung after 4 hours. Ventilation efficiency index and arterial/alveolar oxygen ratios were significantly higher for TLV. CONCLUSIONS: In this lung injury model, both methods achieved equivalent serum gentamicin levels with higher lung levels using PAD during TLV. This study suggests that TLV may provide an effective vehicle for gentamicin delivery in infants with severe pulmonary infection and ventilation/perfusion abnormalities. PMID- 9347000 TI - Elective high-frequency oscillatory ventilation versus conventional ventilation in preterm infants with pulmonary dysfunction: systematic review and meta analyses. AB - OBJECTIVES: To systematically review the evidence to determine whether the routine use of high-frequency oscillatory ventilation (HFOV) as compared with conventional ventilation (CV) is beneficial or harmful in preterm infants requiring mechanical ventilation for pulmonary failure principally due to respiratory distress syndrome. METHODS: All randomized controlled trials of elective HFOV versus CV in preterm infants <36 weeks' gestation with respiratory failure mainly attributable to respiratory distress syndrome were identified from the literature through a search of MEDLINE, EMBASE, Oxford database of Perinatal trials, and previous reviews including cross-references and abstracts. Meta analyses using event rate ratios (ERR), event rate difference, and if significant, number needed-to-treat were calculated (95% confidence limits were used for all analyses). Two prespecified subgroup analyses were performed. RESULTS: Four published trials were included. Meta-analyses revealed the following ERR (95% confidence intervals) for HFOV versus CV: mortality at 28 to 30 days, 1.02 (0.76, 1.39); chronic lung disease (CLD) at 28 days, 0.86 (0.73, 1.01); mortality or CLD, 0.9 (0.80, 1. 01); air-leak syndromes, 1.13 (0.97, 1.33); mechanical ventilation at 28 days, 1.06 (0.84, 1.33); supplemental oxygen at discharge, 0. 59 (0.37, 0.92); intraventricular hemorrhage (IVH) all grades, 1.11 (0.95, 1.29); IVH (grades 3 or 4), 1.32 (1.01, 1.72); and periventricular leukomalacia, 1.39 (0.91, 2.13). In the subgroup of trials in which a high volume strategy (HVS) was used the ERR for CLD was 0.53 (0.36, 0.78); mortality or CLD, 0.56 (0.40, 0.77); supplemental oxygen at discharge, 0.57 (0.36, 0.92); IVH (all grades), 0.90 (0.61, 1.33); and IVH (grades 3 or 4), 0.84 (0.39, 1. 84). Results were similar to these for the trials using surfactant. One recent trial suggests that HFOV may reduce the cost of in-hospital care. CONCLUSIONS: The overall meta analysis is dominated by the HIFI study, which was criticized for its methodology and surfactant was not used. Subsequent studies, most of which used HVS and/or surfactant, have shown benefits in measures of CLD without an increase in rates of IVH. Caution is warranted in interpreting these results because: 1) the treatment is not blinded and this could affect some outcomes; 2) except for one small trial postneonatal survival, lung function, and neurodevelopment have not been reported from HVS trials; and 3) the benefits and disadvantages have not been reported in infants born at different gestational ages or different birth weights. Importantly, results from groups experienced in the use of HFOV may not be readily generalizable. PMID- 9347002 TI - Time for a new growth reference. AB - The National Center for Health Statistics growth reference, recommended by the World Health Organization for international use since the late 1970s, has served many useful purposes. Among the most important are the provision of a single set of growth references for the assessment of the general nutritional status of populations of children in diverse settings, as an ancillary tool to screen children for health and nutrition disorders, and as a basis for educational materials that promote improved child care by families. However, because of serious drawbacks due to the origin and type of data used for their construction and the analytical methods applied in their derivation, the suitability of these curves for international purposes has been challenged recently. PMID- 9347001 TI - Improved oxygenation in a randomized trial of inhaled nitric oxide for persistent pulmonary hypertension of the newborn. AB - OBJECTIVE: To determine the effect of inhaled nitric oxide (NO) on clinical outcome in newborns with persistent pulmonary hypertension (PPHN). DESIGN: A prospective, randomized trial of patients referred to a level 3 nursery in a single large center. Clinicians were not masked to group assignment. Crossover of patients from control to NO treatment was not permitted. METHODS: We randomized 49 mechanically ventilated newborns, transferred to our center with clinical and echocardiographic evidence of severe PPHN (arterial oxygen tension [PaO2] <100; fractional inspired oxygen = 1) to treatment with or without NO. Patients with gestational age <34 weeks or with congenital heart disease or diaphragmatic hernia were excluded. High-frequency oscillatory ventilation was used but not allowed concomitantly with NO. Primary outcome variables were oxygenation, mortality, and use of extracorporeal membrane oxygenation (ECMO). RESULTS: Meconium aspiration syndrome and isolated PPHN were the most common diagnoses (32/49) and were distributed equally between groups. The median age at the time of entry into the study was similar between groups, 25 hours for control patients and 18 hours for NO patients. Median baseline oxygenation index (OI) was similar in 23 control (OI = 29) and 26 NO (OI = 30) patients. Mortality (8%), use of ECMO (33%), median days on mechanical ventilation (9 days), and duration of supplemental oxygen (13 days) were not different between treatment groups. PaO2, oxygen saturation, and OI improved in the NO group compared with baseline and to control patients at 15 minutes. The median percent change in OI (-31%) in the NO group was significantly different from baseline and from the control group. The difference in oxygenation between treatment groups was still apparent 12 hours after baseline. Before cannulation for ECMO, oxygenation was better in the NO group compared with control patients. Among patients who were placed on ECMO, the median time from baseline to ECMO cannulation was 2.4 hours (range, 1 to 12 hours) among control patients and 3.3 hours (range, 2 to 68 hours) for those randomized to receive NO. There was a tendency to observe fewer adverse neurologic events (seizure and intracranial hemorrhage) in the NO group (4/26 vs 8/23). One child with alveolar capillary dysplasia confirmed by postmortem examination could not be weaned from 80 parts per million of NO and transiently developed methemoglobinemia (peak methemoglobin level = 17%). No other side effects were observed. CONCLUSIONS: Although mortality and ECMO use were similar for both treatment groups using this study size and design, sustained improvement in oxygenation with NO and better oxygenation at initiation of ECMO may have important clinical benefits. We speculate that modification of treatment to include specific lung expansion strategies with NO treatment and recognition that early improvement of oxygenation may be sustained with NO may lead to reduced use of ECMO in NO treated patients compared with controls. PMID- 9347003 TI - Lack of deafness in Crigler-Najjar syndrome type 1: a patient survey. AB - We performed a questionnaire survey about 42 patients with Crigler-Najjar syndrome type 1 who were currently alive. Information was obtained on their age, sex, birth weight, gestation, parental consanguinity, other family members affected, age of onset of jaundice, neonatal and postneonatal bilirubin values, neonatal and postneonatal therapy, problems faced with phototherapy, liver transplantation, current growth status, current neurologic status, and the status of hearing. Patients were between 2 months and 21 years of age. There were 18 males and 24 females. Thirty-nine patients had been born at full term gestation and 3 had been preterm. Jaundice was noted on postnatal day 1 in 34%, between days 2 and 4 in 55%, and after day 11 of life in 11% of patients. In the neonatal period bilirubin values (mean +/- SD) were typically 19.8 +/- 4.5 mg/dL. Eighty six percent of patients had neonatal peak bilirubin values of >20 mg/dL. Parental consanguinity was present in 44% and a history of Gilbert's disease in one parent was present in 10% of patients. Causes of exacerbations of jaundice reported were respiratory infections, febrile illnesses, vaccinations, fasting, surgery, emotional stress, and noncompliance with treatment. Neonatal therapy consisted of exchange transfusion in 28%, phototherapy in 79%, phenobarbitone in 20%, and cholestyramine, albumin, infusions, and plasmapheresis in one case each. The mainstay of postneonatal therapy was home phototherapy for 10 to 16 hours, primarily at night during sleep, using blue lights or a combination of blue and fluorescent lights. Some patients used innovatively designed phototherapy units. Problems reported with phototherapy were decreased effectiveness with age, poor compliance, restriction of activity and play, inability to travel or take vacations, irritation from eye shades, difficulty keeping eye protection on, difficulties in temperature maintenance, tanning of the skin, embarrassment from the need to be nearly nude during phototherapy, and difficulty in procuring phototherapy lamps. Other therapies that had been tried included oral agar, albumin infusions, antioxidants, acupuncture, bilirubin oxidase, calcium infusions, clofibrate, cruciferous vegetables, cholestyramine, chlorpromazine, flumecinol, plasmapheresis, tin mesoporphyrin, ursodeoxycholic acid, and urinary alkalinization. Fifteen children had undergone liver transplantation (5 auxiliary and 10 orthotopic). All 42 patients are reportedly of normal height and weight. Neurodevelopmental status is said to be normal in 77% of patients. Two patients have kernicterus, 4 have cerebellar symptoms, and 1 each has developmental delay, mild intention tremor, and mild speech delay. Hearing was reported to be normal in 94% of patients. The 2 children with hearing loss have conductive loss from otitis media. With home phototherapy prolonged, survival free of neurologic deficits is possible in Crigler-Najjar syndrome type 1, but there are many problems associated with phototherapy. Avoiding exacerbations of jaundice is an important aspect of management. Liver transplantation offers the prospect of cure but its risk versus benefit ratio is undetermined. Hearing loss was absent in the patients surveyed despite prolonged exposure to high bilirubin levels, which suggests that bilirubin may not be as ototoxic as is commonly believed. PMID- 9347004 TI - Oral rehydration therapy for diarrhea: an example of reverse transfer of technology. AB - On November 13 and 14, 1996, a scientific symposium on oral rehydration therapy (ORT) was held at the Johns Hopkins University School of Hygiene and Public Health in Baltimore, MD. The purpose of the meeting was to review the current treatment practices for the treatment of this disease in the United States. The group noted that diarrhea resulted in 300 to 400 deaths per year among children, approximately 200 000 hospitalizations, 1.5 million outpatient visits, and costs >$1 billion in direct medical costs. ORT is well established therapy for the treatment and prevention of dehydration due to diarrhea. The principles of ORT treatment include early adequate rehydration therapy using an appropriate oral rehydration solution (ORS), replacement of ongoing fluid losses from vomiting and diarrhea with ORS, and frequent feeding of appropriate foods as soon as dehydration is corrected. The effective use of ORT has saved millions of lives around the world. However, in the United States, ORT is grossly underused. Contrary to the recommendations of the American Academy of Pediatrics (AAP) and the Centers for Disease Control and Prevention (CDC), health care providers overuse intravenous hydration, prolong rehydration, delay reintroduction of feeding, and inappropriately withhold ORT, especially with children who are vomiting. The expert panel noted that the majority of deaths, hospitalization, and visits to emergency departments could be prevented by the appropriate use of ORT. They generated guidelines for the treatment and prevention of dehydration secondary to diarrhea. These measures, together with training providers, could substantially reduce diarrhea mortality and decrease hospitalizations of children by 100 000 per year in the next 5 years. PMID- 9347006 TI - [Unconventional transmissible agents and prion protein: is something still missing?]. AB - Human and animal prion diseases are rare and fatal transmissible neurodegenerative disorders and correspond to a new host-agent interaction. So far, no infectious conventional agent has been isolated from brains of affected. These diseases are characterized by the accumulation in the central nervous system of an abnormal form of the prion protein (PrPc) which is a normal cell component. This abnormal form, PrPSc or PrPres, exhibiting a very high resistance to proteases, results from a conformational change of PrPc and would be an essential part of the transmissible agent, called prion, according to the concept developed by Prusiner since 1982. PrPc is expressed at the cell surface of neurons and peripheral tissues. All mammals studied so far encode PrP. Its tertiary structure has been recently established in part by nuclear magnetic resonance. Although its physiological role is still unknown, PrPc appears to be the receptor of the contaminating agent. Mice devoid of PrP have a normal development and behaviour but are resistant to the disease. Transgenic studies have demonstrated that the molecular basis of the host susceptibility is based in part on the primary structure of PrP. In mouse, sheep and human, different amino acid substitutions in the protein have been associated with opposite incubation times. The propagation of prions would result of a conformational imprinting imposed by infectious PrePres to host PrPc. The newly acquired conformation of the host protein would transmit to all neosynthetized PrPc in a autocatalytic process. Recently, evidence for the existence of different possible conformations of the abnormal protein could explain the variability of the agent strains. Recent data indicate that prion diseases would be the first transmissible disorders in which the information carried by the agent would be hidden in the tertiary structure of a protein and not in a nucleic acid. However, the virus hypothesis has not been totally discarded by some investigators. PMID- 9347007 TI - [Defense proteins in the corneo-conjunctival epithelium]. AB - The eyes are protected by the mechanical action of blinking and the washing action of tears; besides maintaining comfort and serving as the optical surface, the tear film forms the first line of defence. This role is mainly due to specific action of secretory immunoglobulins and continuous presence of unspecific proteins such as lysozyme, lactoferrin and tear lipocalin. PMID- 9347008 TI - [Reflection on 2 current viral diseases: yellow fever and dengue]. AB - Yellow fever and dengue are two current viral diseases induced by flaviviruses and usually transmitted by the same mosquito vector, Aedes aegypti. From 1987 to 1991, 18,753 cases of yellow fever, mainly from Africa, have been notified to WHO, leading to 4,522 deaths. On the other hand, WHO estimates that 2.5 billions individuals living in tropical areas are at risk to contract dengue fevers. In fact, 500,000 patients are hospitalized each year for dengue hemorrhagic fever/dengue shock syndrome and 90% of them are children. Nevertheless, the control of these two viral diseases would be reached easily in destroying mechanically the mosquito larval resting places. Although superficially similar, the two entities are in fact quite different. Relatively few is known about the pathogenesis of yellow fever whereas, for dengue fevers, it is difficult to integrate so many results accumulated to explain the occurrence of haemorrhagic phenomena according to the two main theories so far proposed which are not exclusive. The immunological one (S.B. Halstead) tries to explain the pathological events by the effect of anti-dengue enhancing antibodies acquired during a previous exposure to one of the dengue viruses, whereas that of increased virus virulence (L. Rosen) refers to fast passages between individuals during explosive epidemics. PMID- 9347009 TI - [Endocrine biochemistry of puberty]. AB - Puberty corresponds to the development of gonads and secondary sexual characteristics, and on a biological point of view, to the functional maturation of the gonadal axis. Puberty begins at the age of 11.5 to 12 years in males and 10.5 to 11 years in females. Depending on secondary sexual characteristics, particularly pubic pilosity, puberty is classified in five stages (Tanner's stages). During puberty growth velocity increases in response to gonadal steroid secretion. From a biochemical point of view, three steps are involved in the development of the hypothalamo-hypophysogonadal axis: 1. nocturnal hypothalamic GnRH secretion increases and becomes pulsatile (a peak every 60 to 90 min); 2. the pituitary gonadotrophins FSH and LH follow the same pattern of secretion as GnRH; increase of GnRH and FSH/LH secretion is due to a decrease in hypothalamo hypophysal sensitivity to the negative feed back exerted by circulating gonadal steroids; 3. secretion of estradiol in females and testosterone in males increases, as a consequence of pituitary stimulation. Hormonal exploration of puberty is mainly based on the measurement of FSH-LH and testosterone or estradiol. SDHA is also measured to investigate adrenal androgen secretion, which increases three or four years before puberty; this is related to the maturation of adrenal androgenic function (adrenarche). Dynamic tests are used to evaluate the biological stage of puberty (LH-RH test) and to measure the functional capacity of the testes. Pubertal abnormalities can theoretically be divided into precocious and delayed puberty. In the former, clinical and biological characteristics are used to define: dissociated puberties, central precocious puberty and peripheral precocious puberty. In the latter, hypogonadism has either a central origin (hypogonadotropic hypogonadism) or peripheral origin (hypergonadotropic hypogonadism). PMID- 9347010 TI - [Hair: a powerful biological marker for exposure to xenobiotics]. AB - Human hair analysis is now recognized for evaluating someone exposure to xenobiotics: drugs of abuse, pharmaceuticals and polluants. This paper describes analytical methods than can be used by biologists. For drugs of abuse after decontamination, hydrolysis of hair, selective extraction and derivatization, determinations are performed by gas chromatography-mass spectrometry (GC/MS). Calibration uses deuterated standards. The cut-off value is 0.5 ng/mg for 6 monoacetylmorphine (heroin) and amphetamines and a benzoylecgonine/cocaine ratio > 0.05 for cocaine. Measurement of metabolites from endogenous metabolism sign the exposure (6-monoacetylmorphine and morphine for heroin, benzoylecgonine for cocaine, delta 9-tetrahydrocannabinol carboxylic by-product for cannabis). For drugs, after selective extraction, determinations are performed by GC/MS or liquid chromatography coupled to a diode array detector. Main applications concern drug monitoring to complement blood determinations or when blood collection is missing, as evidence of hidden, illicit or criminal drug exposure. Finally it is a powerfull tool for clinical diagnosis especially when late biological investigations are performed. PMID- 9347011 TI - [Pre-eclampsia and oxygenated free radicals]. AB - Preeclampsia, clinically defined by arterial hypertension, oedema and proteinuria is a frequently occurred complication of pregnancy. This disease seems to be linked to oxidative stress within placenta. The local accumulation of lipid peroxides, resulting from free radicals production increase altered prostacyclin/thromboxane synthesis. Increased production of lipid peroxides, thromboxane and/or cytokines triggered vascular and organic dysfunctions observed in preeclampsia. Changes in lipoprotein metabolism, namely increase in plasma very low density lipoproteins concentrations (VLDL) and oxidized low density lipoproteins (LDL) concentrations could participate to endothelial dysfunctions observed during preeclampsia. PMID- 9347012 TI - [Reference values of apolipoproteins A1 and B. Contribution of international standardization. Travail collaboratif entre la SFBC, l'Arcol et le SFRL]. AB - The utilization of two WHO reference materials, liquid and lyophilized, permitted international standardization of apolipoprotein measurements. We report here the results of a collaborative study between Arcol, SFBC and SFRL in order to establish reference ranges for apo A1 and B on nine standardized systems. A population of 1027 men and women supposed healthy, 4 to 60 year old, have been selected in two Centers for Preventive Medicine. The serum samples were aliquoted frozen at -20 degrees C the day of sampling and analysed by the manufacturers with IFCC standardized calibrants. A specific quality control was performed using a frozen pool of sera. For apo A1, the centile 2.5 of the reference population varies from 1.04 to 1.16 g/l. The range values for the centile 97.5 varies from 1.87 to 2.24 g/l. For apoB, the centile 2.5 varies from 0.43 to 0.57 g/l, and the centile 97.5 from 1.30 to 1.39 g/l. Only one system has a problem of dispersion with an upper limit equal to 1.20 g/l. These results improve that international standardization allowed actually a good comparability of the results, especially for apoB. PMID- 9347013 TI - [Acid-base equilibrium and oxygenation of the human fetus. Study of 73 samples obtained by cordocentesis]. AB - Acid-base state and oxygenation of fetal blood were evaluated in a group of 73 appropriate for gestational age fetuses between 17 and 41 weeks of gestation; fetal blood was obtained by cordocentesis. We measured, on umbilical venous blood, pH, pCO2, bicarbonates and total CO2, pO2 and percent oxygen saturation; we obtained following results: pH = 7.309 +/- 0.054; pCO2 = 5.99 +/- 0.85 kPa; HCO3 = 22.16 +/- 1.90 mmol/l; total CO2 = 22.53 +/- 1.97 mmol/l; pO2 = 6.02 +/- 1.68 kPa; SaO2 = 0.71 +/- 0.18. In comparison with adult normal values, our results report in fetuses a frequent state of mild and mixed acidosis (in about 80% of cases), rather gazeous; pH is significantly correlated with pCO2 (r = 0.799), moreover there is also a metabolic origin in acidosis. We observed progressive modifications of acid-base state and oxygenation in fetal blood with advancing gestational age: decrease of pH and pO2, increase of pCO2 and bicarbonates; these changes appear mainly from 35th week of gestation, except for bicarbonates, which increase regularly during gestational period we have studied. Fetal acid-base state may be explained by physiological differences due to fetal life conditions, in comparison with adult life conditions. Results obtained in this population of normal fetuses can be considered as reference values for studied parameters. PMID- 9347014 TI - [Exploration of immune response in a murine model of congenital toxoplasmosis]. AB - We used a model of acquired toxoplasmosis to study the immune response in pregnant BALB/c mice (IL4+/+) and in pregnant transgenic IL4-deficient BALB/c mice (IL4-/-) during acute toxoplasmosis. Female BALB/c mice were infected orally by 20 tissue cysts of the avirulent PRU strain of Toxoplasma gondii on day 11 of pregnancy. After infection, cultured spleen cells from pregnant mice produced more IFN gamma (a type 1 cytokine) and more NO than non pregnant mice, and the type 2 response (IL4 and IL10) was weak. Although this kind of immune response may be required for mice to recover from toxoplasmosis, pregnant mice were more susceptible to infection than non pregnant mice, as illustrated by a larger parasite load in lungs and brain. Pregnant IL4-/- mice showed lower susceptibility to T. gondii infection and a lower materno-fetal transmission rate (24% versus 53% infected fetus) without increased production of type 1 cytokines (IFN gamma and NO). These data indicate that type 2 response plays an important role in increasing mouse susceptibility to T. gondii infection during pregnancy and that IL4 and pregnancy-associated substances increase the transplacental passage of T. gondii. This is the first time that biased towards type 2 immune response induced by pregnancy was shown to increase susceptibility to T. gondii. PMID- 9347015 TI - [Molecular typing by pulsed field gel electrophoresis of Enterobacter cloacae strains isolated from osteoarticular infections at the Nancy University Hospital 1990-1994]. AB - Enterobacteriaceae represent more than 11% of bacteria involved in osteoarticular infections in adult and, among them, Enterobacter cloacae is found in 12% of the cases. The increased evolution of the antibiotic resistance rate of this species is worrying. However, all isolates responsible for this type of infection at the University Hospital of Nancy from 1990 to 1994 (24 strains isolated from 22 patients presented an identical antibiotype with especially a natural resistance phenotype to beta-lactam antibiotics except one cephalosporinase-over-producing strain. The DNA of these strains was studied by pulse-field gel electrophoresis after digestion by the restriction enzyme XbuI. The great genomic diversity obtained showed that the stability of the antibiotic susceptibility during the period studied was not due to the existence of unique clone but to that of multiple clones. The analysis of the restriction profiles has permitted to achieve a better differenciation of the strains than biotyping and antibiotyping, which confirms the high discrimination power of this genotypic method. PMID- 9347016 TI - [Fetal death in hydrops fetalis at 21 weeks of amenorrhea]. PMID- 9347017 TI - [Influence of bilirubin and turbidity on main determinations performed on the Express 550 analyzer]. PMID- 9347018 TI - [Iron metabolism and HIV infection: preliminary study]. PMID- 9347019 TI - [Antibiotic sensitivity test: contribution to diagnosis, therapy and epidemiology. Travail du comite interface SFBC-SFRL]. PMID- 9347020 TI - [Project of recommandations for the study of monoclonal immunoglobulinopathies in laboratories]. PMID- 9347021 TI - [Association of B-cell lymphoma and T-cell lymphoma in HTLV1 infection]. PMID- 9347022 TI - [Acute diarrhea in a 16-months-old child]. PMID- 9347023 TI - [Quality assurance of the pre-analytic phase: specimen handling]. PMID- 9347024 TI - [9th Session on virology, Versailles. 22-24 May 1997]. PMID- 9347060 TI - The influence of personality, cognition, and behavior on perceptions and metaperceptions following alcoholic beverage selection in a dating situation. AB - This study was designed to investigate perceptions and metaperceptions of individuals in a mixed sex dating situation in which they were offered alcohol. Male and female participants who were unfamiliar with one another were brought together in individual sessions and asked to imagine that they were on a blind date. At the outset of the 15-minute interaction, they each had an opportunity to select an alcoholic or nonalcoholic drink; after receiving their drink, they interacted privately. At the conclusion of the interaction, participants completed questionnaires that solicited their view of their partner and their perception of their partner's view of them. Results indicate that alcohol expectancies predicted alcoholic beverage selection. When women selected alcohol, they found their partner more likeable; those men whose partner was drinking believed that their partner found them more likeable. Additionally, when both partners were drinking they perceived each other as more extroverted, although they did not think their partner perceived them as extroverted. These results are discussed in terms of implications for further research on the role of alcohol as a cue in a mixed-sex social/dating environment. PMID- 9347061 TI - The concurrent validity of a classification of dieters with low versus high susceptibility toward failure of restraint. AB - It has been experimentally shown that the population of high restrained eaters consists of two subpopulations, i.e., those with a low and those with a high susceptibility toward failure of restraint. Only those who combined high restraint with high scores on the disinhibition scale of the TFEQ (Three-Factor Eating Questionnaire) showed overeating after a preload. The aim of the present study was to assess the concurrent validity of a two-factorial classification using the Dutch Eating Behavior Questionnaire (DEBQ) scales for restraint, emotional and external eating, as well as the bulimia scale of the Eating Disorder Inventory (EDI) for locating dieters with low or high susceptibility toward failure. It was examined whether the resulting two-group classification is associated with self-reported behaviors and features of psychopathology, which are generally thought to differentiate both groups of dieters. The results indicated that the two-group classification was indeed associated with many of these behaviors and features of psychopathology. It was concluded that this classification has a good concurrent validity. PMID- 9347062 TI - Association between self-report of cognitive impairment, HIV status, and cocaine use in a sample of cocaine-dependent methadone-maintained patients. AB - HIV-disease as well as chronic cocaine abuse may both produce neuropsychological deficits that could potentially interfere with psychoeducational treatments for drug abuse. In this study, the Neuropsychological Impairment Scale (NIS), a 95 item self-report assessment instrument, was administered to 120 cocaine-dependent methadone-maintained patients (59 HIV-seropositive; 61 seronegative) to assess self-awareness of cognitive deficits in this patient population. HIV-seropositive cocaine users reported significantly more impairment than did HIV-seronegative cocaine users on all summary scores and six of seven clinical subscales. Controlling for the influence of sociodemographic variables (age, sex, ethnicity, and education), acute and chronic cocaine use, and effective distress, there was still a significant relationship between HIV status and self-report of neuropsychological impairment. Relative to patients with known neuropsychological deficits, 41% of HIV-seropositive cocaine users and 31% of HIV-seronegative cocaine users scored in the impaired range on the Global Impairment Index. Implications for treatment are discussed. PMID- 9347063 TI - Inconsistencies in adolescents' self-reports of initiation of alcohol and tobacco use. AB - This study focuses on errors in estimations of age at which alcohol and tobacco are used for the first time. The data come from a 5-year longitudinal study with three measurements. Self-reports about age of first use at the baseline measurement were compared with similar self-reports at two follow-up surveys. Adolescents were more likely to report a higher age of first use at follow-up measurements. Those who at the baseline measurement reported having smoked (n = 338) or consumed alcohol (n = 523) 61.7% and 89%, respectively, underestimated their years of use. By comparison with estimations at the first and third measurement, 13.6% for smoking and 4.6% for drinking were consistent about their age of first use. Self-reports about the age of onset at the baseline measurement were correlated with frequency and intensity of tobacco and alcohol use 5 years later to assess the predictive power of age of onset for later use. With one exception (correlation with intensity of alcohol use 5 years later, r = .14) no significant correlation was found. The results show that the concept age of first use should be utilized with caution for two reasons: (1) the reliability of assessment is insufficient, and (2) correlations of different estimates with actual frequency and intensity of consumption at the third wave are inconsistent. Explanations for errors in measurement, and recommendations for improvement, are discussed. PMID- 9347064 TI - Specific classes of symptoms predict readiness to change scores among dually diagnosed patients. AB - The Transtheoretical Stages of Change model hypothesizes that disadvantages of substance abuse must outweigh advantages before change occurs. This study examined whether substance-related sequelae predicted readiness to change scores. A total of 150 dually diagnosed patients were administered the CAGE questionnaire (CAGE is an acronym for questions about substance use: Cutting down, Annoyed by criticism, feel Guilty, Early morning usage), which was scored for mood and behavior symptoms; a checklist of 12 physical, intrapersonal, and environmental symptoms; and the Brief Readiness to Change questionnaire (RTC). Regression analyses suggested that more physical and mood symptoms were predictive of higher total RTC (R2 = .11); physical, mood, and behavioral symptoms were predictive of higher contemplation scores (R2 = .17), whereas fewer physical symptoms were predictive of higher precontemplation scores (R2 = .05). The results suggest that the relative severity of physical, mood, and behavior symptoms may be important factors related to the contemplation of change among dually diagnosed patients. PMID- 9347065 TI - What is binge eating? A comparison of binge eater, peer, and professional judgments of eating episodes. AB - Binge eating is a central diagnostic feature of bulimia nervosa (BN) and binge eating disorder (BED), yet the phenomenon of bingeing has not been adequately defined, and there is considerable variability in how individuals label eating episodes as binges. We examined the agreement among binge-eating individuals, non eating-disordered peers, and professional dietitians over whether particular eating episodes were binges. Twenty-nine females with BED, fifteen nonclinical binge eaters (NCB), three peer judges, and three dietitians rated a sample of eating episodes of the binge-eating individuals as either binges or nonbinges based on the types and amounts of food eaten as well as the duration of each eating episode. BED participants labeled a significantly higher proportion of their intakes as binges relative to NCB participants. Peer judges were more likely than were dietitians to label participants' eating episodes as binges. Agreement within dietitian and peer groups was poor to fair, whereas agreement between these groups was fair. Finally, agreement between participants and the external judges (i.e., peers, dietitians) was poor. Possible explanations for these findings as well as implications for the diagnosis of BN and BED are discussed. PMID- 9347066 TI - Prevalence and correlates of heavy smoking in Vietnam veterans with chronic posttraumatic stress disorder. AB - A study was conducted to investigate smoking patterns in 445 Vietnam veterans with and without posttraumatic stress disorder (PTSD). Combat veterans with PTSD reported similar occurrence of smoking (53%) compared to combat veterans without PTSD (45%). For those who smoked, combat veterans with PTSD reported a significantly higher rate of heavy smoking (> or = 25 cigarettes daily): 28% of combat veterans without PTSD were heavy smokers and 48% of combat veterans with PTSD were heavy smokers. PTSD diagnosis and heavy smoking status were independently and differentially related to motives for smoking. In combat veterans with PTSD, heavy smoking status was positively related to total health complaints, lifetime health complaints, health complaints in the past year, negative health behaviors, total PTSD symptoms, DSM-IV C cluster (avoidance and numbing) and D cluster (hyperarousal) PTSD symptoms. Heavy smoking status was also associated with fewer positive health behaviors. PMID- 9347067 TI - Childhood hyperactivity, gender, and Cloninger's personality dimensions in alcoholics. AB - Cloninger described a classification system for subtyping alcoholics based on personality dimensions of novelty seeking (NS), harm avoidance (HA), and reward dependence (RD). The relationship between these dimensions and childhood hyperactivity was examined. Mild to moderate alcohol-dependent adults (68 male and 34 female) were administered Tarter's HK/MBD Childhood Symptom Checklist, Cloninger's Tridimensional Personality Questionnaire, and the Beck Depression Inventory. Alcoholics with childhood hyperactivity (38% of sample), regardless of gender, exhibited greater NS than did alcoholics without childhood hyperactivity. Female alcoholics scored significantly higher on RD compared to males. Findings suggest that high NS, proposed by Cloninger as a predominantly male feature, is related to childhood hyperactivity, independent of gender. PMID- 9347068 TI - Development and validation of the drug-taking confidence questionnaire: a measure of coping self-efficacy. AB - The Drug-Taking Confidence Questionnaire (DTCQ; Annis & Martin, 1985) is a 50 item self-report questionnaire developed to assess situation-specific coping self efficacy for use of a particular substance of abuse (e.g., cocaine, heroin, alcohol, cannabis, etc.). Results from exploratory and confirmatory factor analyses of the DTCQ on 713 clients seeking treatment at an addiction treatment facility provided strong evidence for the situation-specificity of efficacy beliefs. An 8-factor first-order model, based on the eight high-risk categories for relapse identified by G.A. Marlatt (Marlatt & Gordon, 1980) and a 3-factor second-order model (i.e., negative situations, positive situations, and temptation situations) provided the best fit to the data. All eight subscales of the DTCQ were shown to have good reliability (alphas .79 to .95). Extensive convergent and discriminant validity analyses for the DTCQ and its subscales in relation to ADS, DAST, OES, DRIE, SCQ, SCL-90R, BDI, HS, and SOCRATES substantiate that the DTCQ is a promising tool for further research and clinical application. PMID- 9347069 TI - Exhaled carbon monoxide and urinary cotinine as measures of smoking in pregnancy. AB - We examined the relationships among self-reported cigarette consumption, exhaled carbon monoxide, and urinary cotinine/creatinine ratio in pregnant women. Information on these measures of smoking was collected at first and 36th week prenatal visits. Correlations between cigarette consumption and exhaled carbon monoxide were .65 at the first visit and .70 at the 36th-week visit. For urinary cotinine/creatinine ratio, the correlations were .61 and .65, respectively, at these visits. Correlations with change in cigarette consumption between the two visits were .37 for change in carbon monoxide and .33 for change in urinary cotinine/creatinine ratio. Urinary cotinine/creatinine ratio had slightly higher overall agreement with self-reported smoking status and was less likely to misclassify smokers than carbon monoxide. We conclude that urinary cotinine/creatinine ratio is the more accurate measure for validating smoking status among pregnant women, but exhaled carbon monoxide is the better measure of cigarette consumption and of changes in consumption. PMID- 9347070 TI - Initial and experimental stages of tobacco and alcohol use during late childhood: relation to peer, parent, and personal risk factors. AB - A staged model of smoking adoption has been widely applied in studies of adolescent smoking. The present study applied this model to examine the preliminary stages of tobacco and alcohol use by children. Using discriminant analysis, factors associated with the abstinence, initiation, and experimentation stages of tobacco and alcohol use were compared in a sample of 1,272 children in grades 4 and 6. Modeling of use by best friends and the perceived prevalence of use among same-age peers were most strongly related to the initiation and experimentation stages of tobacco and alcohol use. Other key factors were offers from parents and friends, adjustment to school, and behavioral self-regulation. The weakest factors were parental modeling and self-esteem. The initiation and experimentation stages are not as highly differentiated among children as other studies have found them to be among adolescents, suggesting that if initiation occurs during childhood, progression to experimentation is likely. Prevention programs could simultaneously influence children's risk of tobacco and alcohol use by targeting the common risk factors for preliminary use of these substances. PMID- 9347071 TI - Bulimia as a disturbance of narcissism: self-esteem and the capacity to self soothe. AB - A review of the literature on eating disorders reveals that, although psychodynamic formulations linking narcissistic dynamics--particularly difficulties with self-soothing--and eating disorders are common in the theoretical and clinical literature, little empirical work has attempted to substantiate this claim. In this study, 117 women completed the Eating Disorder Inventory and the Bulimia Test Revised and four scales that measure different components of narcissism (the Multidimensional Self-Esteem Inventory, measuring self-esteem, the Self-Care Questionnaire, and two subscales of the Ego Functioning Assessment Questionnaire, measuring self-soothing). The four scales used to assess narcissism were all highly correlated with each other, indicating that they measure a similar construct. In addition, the eating-disorder measures were correlated with the measures of narcissism, suggesting that a relationship exists between bulimia and narcissism, as assessed using self-report instruments. PMID- 9347072 TI - A comparison sample validation of "your workplace": an instrument to measure perceived alcohol support and consequences from the work environment. AB - This paper addresses the psychometric properties of Your Workplace (YWP), an instrument developed to measure perceived influence of workplace norms and attitudes on alcohol involvement and the experience of adverse consequences. Data were collected from a large, geographically dispersed sample of aftercare and outpatients recruited for a multisite clinical trial of alcoholism treatments, Project MATCH. Administration of YWP at the baseline assessment was restricted to workforce participants. A confirmatory factor analysis addressed instrument structure. Internal consistency and concurrent association between measures of general social support, alcohol specific support, and alcohol involvement were examined. YWP scales were found to have adequate internal consistency reliability. Correlation between YWP scales and concurrent measures of alcohol involvement were among the strongest found. Identification of workplace influences on alcohol involvement allows refined assessment and fosters a comprehensive approach to treatment of alcoholism. PMID- 9347073 TI - Effects of chronic ethanol consumption and aging on 5-HT2A receptors and 5-HT reuptake sites. AB - The serotonergic system in brain is adversely affected by both aging and chronic ethanol consumption. The present study examined the combined effects of aging and chronic ethanol consumption on two components of the serotonergic system. Serotonin (5-HT) reuptake sites and 5-HT2A receptors were quantitated in brain areas of 5-, 14-, and 24-month-old male Fischer 344 rats that were pair-fed a control or 6.6% (v/v) ethanol-containing liquid diet on a chronic basis. The regions examined include those containing the cell bodies and projections of serotonergic neurons. These experiments demonstrated the sensitivity of the serotonergic system of male Fischer 344 rats to both aging and chronic ethanol consumption. In control rats, aging was associated with a decline in the concentration of 5-HT2A receptors in the nucleus accumbens and four cortical regions: frontal, parietal, piriform, and cingulate cortex. 5-HT2A receptors were also reduced in the frontal, parietal, and cingulate cortex of aged ethanol-fed rats. In contrast, 5-HT reuptake sites were increased in older rats in the frontal cortex, nucleus accumbens, amygdala, and CA3 region of the hippocampus. If comparable changes in 5-HT2A receptors and 5-HT reuptake sites occur in elderly humans, they may contribute to ethanol consumption, and lead to cognitive and other age-related problems. These changes may also alter the effectiveness of serotonergic drugs used in the treatment of alcoholism and mental disorders. The effects of chronic ethanol consumption were more limited. The only significant ethanol effect was an increase of 5-HT2A receptors in the nucleus accumbens of 5 month-old ethanol-fed rats. PMID- 9347074 TI - Moderate alcohol consumption does not augment bone density in ovariectomized rats. AB - Moderate levels of alcohol consumption have been reported to have a beneficial effect on bone mineral density in postmenopausal women. The objective of this study was to examine the effect of a moderate level of alcohol consumption on bone density in a rigorously controlled animal model of osteoporosis. Ovariectomized and nonovariectomized rats were placed on standard lab pellets with free access to deionized water ad libitum. Alcohol-treated animals were given 0.38 g/kg of alcohol daily by intubation in the mid-afternoon and free access to standard lab pellets for 6 weeks. The amount of the alcohol solution was calculated daily to give the human equivalent of 2 glasses of wine/day. Pair fed control animals were given, on the following day, an equal volume of the diet consumed by individual ethanol-fed rats. They received daily intubation solutions, with the ethanol replaced by isocaloric and isovolumetric amounts of maltose-dextrin. Chow-fed control animals received no intubations and were given access to standard lab pellets ad libitum. Ovariectomized animals had increased weight and decreased femur density and bone volume per total volume. They also had decreased total trabecular area, trabecular area, and number, as well as increased trabecular separation. Significant differences were found between the ovariectomized and nonovariectomized animals in the parameters under discussion, but there were no differences between diet groups. No beneficial effects were found after daily alcohol treatments. PMID- 9347075 TI - Effects of maternal ethanol consumption and buspirone treatment on 5-HT1A and 5 HT2A receptors in offspring. AB - In utero ethanol exposure results in a decreased concentration of serotonin (5 HT) in brain regions containing the cell bodies of 5-HT neurons and their cortical projections. The concentration of 5-HT reuptake sites is also reduced in several brain areas. The present study extended prior work by evaluating the effects of chronic maternal ethanol consumption and maternal buspirone treatment on 5-HT1A and 5-HT2A receptors in multiple brain areas of offspring. Receptors were quantitated early in postnatal development and at an age when the 5-HT networks are normally well-established. Because fetal 5-HT functions as an essential neurotrophic factor, these studies also determined whether treatment of pregnant rats with buspirone, a 5-HT1A agonist, could overcome the effects of the fetal 5-HT deficit and prevent ethanol-associated receptor abnormalities. The results demonstrated that in utero ethanol exposure significantly alters the binding of 0.1 nM [3H]-8-hydroxy-dipropylaminotetralin to 5-HT1A receptors in developing animals. Ethanol impaired the development of 5-HT1A receptors in the frontal cortex, parietal cortex, and lateral septum; these receptors did not undergo the normal developmental increase between postnatal days 19 and 35. The dentate gyrus was also sensitive to the effects of in utero ethanol exposure. 5 HT1A receptors were increased in this region at 19 days. Maternal buspirone treatment prevented the ethanol-associated abnormalities in 5-HT1A receptors in the dentate gyrus, frontal cortex, and lateral septum. Neither maternal ethanol consumption nor buspirone treatment altered the binding of 2 nM [3H]ketanserin to 5-HT2A receptors in the ventral dentate gyrus, dorsal raphe, parietal and frontal cortexes, striatum, substantia nigra, or nucleus accumbens. PMID- 9347076 TI - Ethanol consumption suppresses cell-mediated inflammatory responses and increases T-helper type 2 cytokine secretion in Trichinella spiralis-infected rats. AB - Alcohol consumption has been shown to be associated with immune suppression and immune modulation. In this study, the effects of ethanol ingestion on the host immune responses to Trichinella spiralis infection and the subsequent secretion of T-helper cell-associated cytokines were investigated in rats. At the early phase of T. spiralis infection, ethanol-fed animals showed decreased numbers of blood neutrophils and eosinophils, and a decreased secretion of interferon-gamma (IFN-gamma) by mesenteric lymph node cells, compared with pair-fed controls. Suppression of this early inflammatory response to infection in the ethanol treated groups resulted in a slower rate of expulsion of intestinal adult worms and a higher fecundity rate for female worms, compared with pair-fed controls. A dramatic decrease in blood neutrophils in the ethanol-treated groups was also manifested on day 9 postinfection. At that time, mesenteric lymph node cells from the ethanol-treated groups secreted higher amounts of mast cell proliferation enhancing activity, which has been shown to contain T-helper type 2-associated cytokines. At the later phase of infection (day 12 to 20 day postinfection), ethanol-treated animals contained higher numbers of blood eosinophils and secreted an increased amount of interleukin-5 and mast cell proliferation enhancing activity, compared with pair-fed controls. Although there was a slight rise with time after infection, the level of serum corticosterone was not significantly increased in the ethanol groups. Therefore, it is not likely that the observed immune modulations caused by ethanol consumption, especially in the early phase of infection, is the effect of elevated levels of corticosterone in the circulation. The present study found that ethanol consumption suppressed the initial amount of interferon-gamma secretion and inflammatory response, and may have directly or indirectly led to an enhancement of the secretion of T-helper type 2-type cytokines later in the primary immune response to T. spiralis infection. PMID- 9347077 TI - Differential sensitivity of human neuroblastoma cell lines to ethanol: correlations with their proliferative responses to mitogenic growth factors and expression of growth factor receptors. AB - Early ethanol exposure depletes neurons in the developing nervous system, however the effects on neuronal precursors are not homogeneous. Some cells are more susceptible to ethanol toxicity than others. Growth factors are important mitogens for neuronal precursors. We tested the hypothesis that the differential sensitivity of neuronal precursors to ethanol is determined by their responses to growth factors using an in vitro model (SH-SY5Y, SK-N-SH, and IMR32 neuroblastoma cells) of neuronal precursors. The three cell lines were raised in a medium containing 10% or 0% fetal calf serum. Cells were exposed to ethanol and/or a growth factor. These factors included basic fibroblast growth factor, epidermal growth factor, insulin-like growth factor-I, nerve growth factor, and platelet derived growth factors AA and BB. The numbers of cells per culture were counted both before and after 3 days of ethanol and/or growth factor treatment. In addition, the effect of ethanol exposure on the expression of receptors for these growth factors was examined. Neuroblastoma cells displayed differential sensitivity to ethanol. The growth of SH-SY5Y and SK-N-SH cells was inhibited by ethanol in a concentration-dependent manner. Ethanol did not affect cell viability. Thus, this inhibition resulted from a reduction of cell proliferation. In contrast, IMR32 cells were not affected by ethanol (even at concentrations as high as 800 mg/dl). The response to growth factors was also heterogeneous. In serum-supplemented medium, SH-SY5Y and SK-N-SH cells were stimulated by all of the tested growth factors. For cells raised in a serum-free medium, only the nerve growth factor was ineffective. IMR32 cells, however, were unaffected by most of these growth factors, regardless of the medium conditions. Ethanol blocked the action of all growth factors tested. In general, all cells expressed the specific receptors for the six growth factors. Only the expression of the basic fibroblast growth factor, insulin-like growth factor-I, and nerve growth factor receptors were reduced by ethanol exposure. In summary, neuroblastoma cells exhibit differential susceptibility to ethanol, and this correlates with their response to mitogenic growth factors. Some growth factors are a target of ethanol toxicity. These heterogeneous effects seem to parallel ethanol-induced changes of proliferating neuronal precursors in vivo. PMID- 9347078 TI - Hormonal (ACTH, cortisol, beta-endorphin, and met-enkephalin) and cardiovascular responses to hyperthermic stress in chronic alcoholics. AB - Chronic alcohol drinking causes profound alterations in hypothalamic-pituitary function. In the present study, endocrine [corticotropin (ACTH), beta-endorphin, cortisol, and met-enkephalin] and cardiovascular (blood pressure) changes in response to hyperthermic stress (sauna at 90 degrees C for 30 min) were evaluated in 25 normal men (25 to 50 years old) and in 48 male alcoholic subjects (34 to 56 years old) after 5 weeks of abstinence. Significantly lower increments in systolic blood pressure were observed in alcoholics than in control subjects. Furthermore, alcoholics showed lower ACTH, beta-endorphin, and cortisol increments in response to sauna than normal controls. In contrast, sauna-induced hyperthermia did not change significantly the circulating met-enkephalin levels in either normal controls or chronic alcoholics. These data suggest that an impairment in the adaptive response to stress affects alcoholic men even after a few weeks of abstinence from alcohol. PMID- 9347079 TI - Activation of arachidonic acid-specific phospholipase A2 in human neuroblastoma cells after chronic alcohol exposure: prevention by GM1 ganglioside. AB - Human neuroblastoma cells were exposed to ethanol (EtOH; 100 mM) in culture for various time periods. It was found that chronic EtOH exposure increased the arachidonyl-specific phospholipase A2 (PLA2) activity significantly in both cytosol (1.6-fold) and membrane (2.2-fold) fractions when 1-palmitoyl-2 arachidonyl-sn-glycero-3-phosphocholine was used as a substrate. This arachidonyl specific PLA2 activity progressively increased with increasing duration of EtOH exposure and reached peak level at 72-hr EtOH exposure (chronic). A significant amount of the PLA2 activity was associated with the membrane fraction. No significant difference in PLA2 activity was observed when 1-palmitoyl-2 oleoyl or linoleoyl-sn-glycero-3-phosphocholine was used as a substrate. It was also found that co-treatment of neuroblastoma cells with ganglioside GM1 reduced the EtOH induced activation of arachidonyl-specific PLA2 activity. The present results indicate that arachidonic acid-specific PLA2 may play a role in adaptation mechanisms to chronic EtOH in cultured neuroblastoma cells. Ganglioside GM1, in part, may exert its neuroprotective effects by modulating arachidonyl-specific PLA2 activity in chronic EtOH-exposed neuroblastoma cells. PMID- 9347080 TI - Collagen synthesis by liver stellate cells is released from its normal feedback regulation by acetaldehyde-induced modification of the carboxyl-terminal propeptide of procollagen. AB - Acetaldehyde stimulates collagen synthesis in stellate cells and forms adducts with procollagen in the liver of alcoholics. To assess the possibility that modification of the carboxyl-terminal propeptide by acetaldehyde affects its capacity to exert a feedback inhibition of collagen synthesis after splitting from procollagen, the propeptide was prepared by gel filtration of the bacterial collagenase digests of procollagen type I (obtained from 10(9) calvaria fibroblasts of newborn rats) and reacted with either 250 mM acetaldehyde and 100 mM CNBH3 or with 170 microM acetaldehyde without reducing agents, to mimick in vivo conditions. The unmodified propeptide produced a concentration-dependent inhibition of collagen synthesis by Ito cells. By contrast, the acetaldehyde modified propeptide produced a lesser inhibition of procollagen synthesis in the cells, associated with a greater accumulation of collagen in the media. The incubation with 170 microM acetaldehyde and, to a lesser extent, 50 mM ethanol produced collagenase-digestible adducts in stellate cells. Thus, the formation of acetaldehyde adducts with the carboxyl-terminal propeptide of procollagen may account, at least in part, for the stimulatory effect of acetaldehyde on collagen synthesis by stellate cells and may lead to collagen accumulation through a decrease of the normal feedback regulation of collagen synthesis by the propeptide. PMID- 9347081 TI - Ethanol feeding inhibits proinflammatory cytokine expression from murine alveolar macrophages ex vivo. AB - The prolonged and excessive consumption of alcohol has been shown to predispose the host to a variety of infectious complications, which may be due, in part, to the inability to produce important activating and chemotactic cytokines. In this study, we assessed the effect of alcohol ingestion on the expression of tumor necrosis factor-alpha (TNF-alpha), and the chemokines macrophage inflammatory protein-2 (MIP-2) and macrophage inflammatory protein-1 alpha (MIP-1 alpha) from murine alveolar macrophages (AMs) cultured ex vivo. Two-week ethanol feeding resulted in substantial impairment in the lipopolysaccharide (LPS)-induced expression of TNF-alpha, MIP-2, and MIP-1 alpha mRNA, and protein from LPS stimulated AMs, compared with cytokine production from AMs obtained from CD-1 mice receiving an isocaloric control diet. These findings indicate that ethanol feeding results in diminished production of chemotactic and/or activating cytokines from AMs ex vivo that may contribute to the impairment in lung inflammatory responses and antimicrobial host defense that is observed in the setting of alcohol ingestion/intoxication clinically and experimentally. PMID- 9347082 TI - MK-801 administration during ethanol withdrawal in neonatal rat pups attenuates ethanol-induced behavioral deficits. AB - Alcohol exposure during development can produce central nervous system dysfunction, resulting in a wide range of behavioral alterations. The various mechanisms by which alcohol causes these behavioral changes, however, remain unknown. One mechanism that has been suggested is NMDA receptor-mediated excitotoxic cell death produced by ethanol withdrawal. The present study examined whether MK-801, an antagonist of the NMDA receptor that has been shown to protect against NMDA receptor-mediated excitotoxicity, could block alcohol's adverse effects on behavior. Sprague-Dawley rat pups were exposed to alcohol (6.0 g/kg) in a binge-like manner on postnatal day 6 using an artificial rearing procedure. Subjects then received an injection of MK-801 (0.1 mg/kg) or vehicle during withdrawal, 21 hr after ethanol exposure. At postnatal day 40, all subjects were tested on a serial spatial discrimination reversal task. Ethanol-exposed subjects were impaired in both discrimination and reversal learning, and committed a significantly greater number of perseverative-type errors, compared with controls. MK-801 administration during ethanol withdrawal significantly attenuated ethanol-induced deficits in reversal learning and increases in perseverative-type errors, whereas MK-801 exposure by itself had no significant effect on performance. Thus, exposure to MK-801 during ethanol withdrawal partially protected against alcohol-related disruptions in spatial reversal learning. These results support the suggestion that NMDA receptor-mediated excitotoxicity may be one mechanism by which alcohol induces behavioral teratogenicity. PMID- 9347083 TI - Alterations in tumor necrosis factor-alpha, interferon-gamma, and interleukin-6 production by natural killer cell-enriched peripheral blood mononuclear cells in chronic alcoholism: relationship with liver disease and ethanol intake. AB - No previous studies have been reported on human alcoholism in which the pattern of cytokine secretion by natural killer (NK) cells is explored. The goal of the present study was to evaluate the role of NK cells in the production of cytokines in patients with chronic alcoholism, analyzing at the same time the possible relationship between cytokine production and both alcoholic liver disease and ethanol (EtOH) intake. A total of 30 chronic alcoholic patients-11 without liver disease [alcoholics without liver disease (AWLD) group] and 19 diagnosed of alcoholic liver cirrhosis-were included in this study. Twenty-five age- and sex matched healthy volunteers were analyzed as controls. Production of interferon (IFN)-gamma, tumor necrosis factor-alpha (TNF-alpha), and interleukin (IL)-6 was performed on NK-enriched peripheral blood mononuclear cells (PBMC) after stimulation with IL-2 and IFN-alpha. In AWLD patients, the production of TNF alpha was significantly reduced, compared with normal controls, under both IFN alpha (p < 0.01) and IL-2 (p < 0.05) stimulation. In patients with cirrhosis, TNF alpha production by PBMC enriched in NK cells varied depending on the EtOH intake status at the moment of evaluation. Accordingly, an increased concentration of this cytokine was detected in the supernatants of cirrhotic patients and active EtOH intake, particularly after IFN-alpha stimulation (p < 0.05); whereas, in patients with at least 1 year of alcohol withdrawal, TNF-alpha levels remained within normal range. The results on the production of IL-6 and IFN-gamma in AWLD and cirrhotic patients showed that only cirrhotic patients with a prolonged EtOH withdrawal period display abnormal production. Accordingly, in this group of patients, a significantly increased release of IL-6 was observed after both IFN alpha and IL-2 stimulation (p < 0.01 and p < 0.05, respectively). By contrast, a lower IFN-gamma production (p < 0.005) was detected with respect to the control group. Our results point to the existence of an abnormal cytokine secretion by NK cells from chronic alcoholism patients, which depend on both the existence of liver disease and the status of EtOH intake. PMID- 9347084 TI - Ultrasonic vocalization behavior differs between lines of ethanol-preferring and nonpreferring rats. AB - To further understand the relationship between emotional state and alcohol intake in rats, the tendency to emit ultrasonic vocalizations in response to an aversive, but nonpainful, air puff stimulus was tested in several rat lines. Included in this group were Maudsley Reactive (MR) and Non-Reactive (MNR) rats, and several lines of rats with either high ethanol preference or a low ethanol preference: Preferring, (P), Alko-Alcohol (AA), and Fawn-Hooded (FH) animals; and Non-Preferring (NP), Alko-Non-Alcohol (ANA), and Flinders Resistant Line (FRL). MR rats emitted fewer ultrasonic vocalizations (USVs) and showed less preference for ethanol than did MNR animals. An overall analysis that included the P, NP, FH, FRL, AA, and ANA groups demonstrated a significant negative correlation between the total number of USVs emitted and ethanol consumption. NP, FRL, and especially ANA rats (low ethanol-preferring) emitted the most USVs--to an extent similar to that typically found for normal rats. The duration of vocalizing was higher only in the NP and the FRL rats the relative to their P and FH comparison groups, respectively. In the ethanol-preferring and nonpreferring lines, the numbers of USVs emitted correlated positively with the duration of vocalizing, but not with the latency to vocalize, which in turn did not correlate strongly with ethanol intake. The latency to vocalize did not correlate significantly with ethanol intake across all drinking lines or MR or MNR rats, but was found to be higher in FH and AA rats relative to their nondrinking comparison groups. These associations suggest that the relationship between emotional state and ethanol drinking is complex and cannot be attributed to a simple elevated state of anxiety or emotionality. Further examination of the central nervous system mechanisms mediating the difference in USVs between paired lines of ethanol preferring and nonpreferring rats may identify neurochemical factors that predict ethanol preference. PMID- 9347085 TI - Transglutaminase activity in rat brain after ethanol exposure. AB - The effect of acute and chronic ethanol treatment on the activity of tissue transglutaminase (a calcium-dependent enzyme that catalyzes the covalent association between proteins, as well as proteins and polyamines) was studied in homogenate and in the cytosolic fraction of rat brain (telencephalon and diencephalon). A single dose of ethanol (5 g/kg of body weight, by gastric intubation) caused a 2-fold increase in enzyme activity at 6 hr after the ethanol dose, with a return toward the basal values at 24 hr. In vitro experiments with ethanol or acetaldehyde showed that the increase in transglutaminase activity was due to ethanol per se and not to its metabolism. The enzyme stimulation was correlated with a decrease in the levels of the polyamine spermine, a physiological substrate for the enzyme. Similar results were also found in the brain from rats fed on an ethanol diet for 4 months. The enhancement in tissue transglutaminase activity may thus lead to a decline in spermine, a polyamine known to have important protective functions in the cell. PMID- 9347086 TI - Ethanol inhibits inducible nitric oxide synthase transcription and post transcriptional processes in vivo. AB - The effects of ethanol (ETOH) on post-transcriptional regulation of inducible nitric oxide synthase (iNOS) in vivo has not been demonstrated. We examined the effect of ETOH on iNOS mRNA, protein, and the production of the nitrate and nitrite anion (RNI) in rat lung alveolar macrophages (AM) in vivo when stimulated by lipopolysaccharide (LPS) and dibutyryl cyclic AMP (DB-cAMP). Sprague-Dawley rats (225-250 g) (n = 5-7/gp) were given intratracheal LPS (0.6 mg/kg) or DB-cAMP (0.1 and 1 mg/kg) 30 min after ETOH (4 mg/kg, intraperitoneally (ip)) or phosphate-buffered sterile saline (PBS) (5 ml/kg, ip) or pyrrolidine dithiocarbamate (PDTC 10 mg/kg, intratracheally) or 15 min after diethyl dithiocarbamate (DETC) (5 mg/kg, intratracheally). At selected times after administration of LPS or DB-cAMP, the animals were anesthetized, the lungs with the heart attached were removed and the lungs subjected to bronchoalveolar lavage (BAL). The BAL fluid was assayed for tumor necrosis factor alpha (TNF alpha) and RNI. The BAL fluid AM were isolated and analyzed for iNOS mRNA and protein with competitor-equalized polymerase chain reaction (PCR) and Western blots, respectively. The ex vivo incubates of AM were assayed for RNI and TNF alpha. LPS and DB-cAMP each increased iNOS mRNA and protein in AM and RNI in the BAL fluid and ex vivo incubates of AM. However, the peak of the increase of iNOS mRNA occurred at 2 hr for DB-cAMP and 4 to 6 hr for LPS. Only LPS increased the concentrations of TNF alpha in BAL fluid and ex vivo incubates of AM. ETOH attenuated LPS-mediated up-regulation of iNOS mRNA and TNF alpha and iNOS protein and RNI produced by the AM. In contrast, pretreatment of rats with ETOH did not affect DB-cAMP-mediated increases of iNOS mRNA in AM at any time but suppressed the amount of iNOS protein and RNI produced in DB-cAMP-stimulated AM. DETC, but not PDTC, attenuated LPS-mediated up-regulation of iNOS mRNA without effects on that produced by DB-cAMP. Since ETOH and DETC, but not PDTC, suppressed LPS mediated but not DB-cAMP-mediated transcription of iNOS, we conclude that two distinct pathways exist for induction of iNOS mRNA by these agonists. ETOH and DETC may inhibit the LPS-mediated activation pathway by acting as antioxidants. Also, since ETOH inhibited DB-cAMP-mediated increases in iNOS protein without affecting iNOS mRNA ETOH also acts at a post-transcriptional or translational site to inhibit iNOS protein in rat AM in vivo. PMID- 9347087 TI - Therapeutic motor training increases parallel fiber synapse number per Purkinje neuron in cerebellar cortex of rats given postnatal binge alcohol exposure: preliminary report. AB - Because therapeutic approaches to fetal alcohol effects in humans have been rare, this study explored the rehabilitative effect of complex motor training on an animal model of binge drinking in the third trimester of human pregnancy. Neonatal alcohol exposure induces significant and permanent reductions in Purkinje and granule cell number accompanied by impaired motor behavior in rats. The purpose of this study was to determine: (1) whether the motor skill impairment caused by exposure to alcohol in the early postnatal period could be ameliorated by the learning of a set of complex motor tasks that had been demonstrated to cause synaptogenesis in the cerebellar cortex; and (2) the extent to which cerebellar neurons in alcohol-exposed (AE) rats exhibit synaptic plasticity. The AE group was given 4.5 g/kg/day of ethanol from postnatal days 4 to 9 via an artificial rearing procedure producing a mean peak blood alcohol level of 257 mg/dl. Control groups consisted of a gastrostomy control (GC) group, that received an isocaloric mixture of maltose/dextrin instead of ethanol, and a suckle control (SC) group, that was reared normally by dams. At approximately 6 months of age, animals from the three groups were assigned either to a rehabilitation condition (RC; that received 10 days of training on the motor tasks) or to an inactive condition (IC; where rats stayed in isolation in their cages). Although SC rats were significantly faster to complete the course in the first 5 days of training, there were no differences in ability to perform among animals from all three groups-SC, GC, and AE--at the end of the training period. Unbiased stereological techniques were used to obtain estimates of the number of parallel fiber synapses/Purkinje cell within the cerebellar paramedian lobule. Results showed that the RC rats from the SC and AE groups had significantly more synapses/Purkinje cell than corresponding IC animals. These data demonstrate that rehabilitative intervention (complex motor training) can improve motor performance impaired by postnatal alcohol exposure and that surviving Purkinje neurons retain the capacity for synaptic plasticity. PMID- 9347088 TI - The nexus between alcohol and violent crime. AB - The present study examines the nexus between alcohol and violent crime by specifying alcohol as a moderating variable that may interact with other major causes of violent crime. Four major causes of violent crime at the individual level are identified: deviant motives or attitudes, aggression and hostility, impulsivity, and problem-solving ability. Analyses are conducted at two levels of aggravated assault: prevalence of assault and frequency of assault. At the level of prevalence of assault, data indicate that the usual drinking pattern does not constitute an independent cause, but has significant interactions with two of the major causes: deviant attitudes and aggression and hostility. However, in the analysis of the frequency of assault, the findings indicate a pattern that both usual drinking pattern and drinking before offending have independent explanatory power for aggravated assault, but no interactions were found. These findings suggest that alcohol may have different roles in explaining different levels of violent offending. PMID- 9347089 TI - Polymorphism of ADH and ALDH genes among four ethnic groups in China and effects upon the risk for alcoholism. AB - The alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) that metabolize ethanol are polymorphic. Different alleles encode subunits of the enzymes that differ in their rate of metabolizing ethanol. These polymorphisms are distributed differently among populations and have been shown to influence the risk for alcoholism in some Asian populations. We have examined the allele frequencies at the ADH2, ADH3, and ALDH2 loci in four populations from China (Han, Mongolian, Korean, and Elunchun) and in alcoholics within each population. The four populations differ in allele frequencies, with the Elunchun having a much lower frequency of ADH2*2 alleles, and the Mongolian and Elunchun having a much lower frequency of ALDH2*2 alleles. Within each population, alleles at one or more of these three loci are protective against alcoholism, although the populations differ in which loci play significant roles. The protective allele at each locus (ALDH2*2, ADH2*2, and ADH3*1) encodes a subunit that either metabolizes ethanol to acetaldehyde more rapidly or slows the conversion of acetaldehyde to acetate. Taken as a whole, data demonstrate that genetic differences in the enzymes that metabolize alcohol can substantially affect the risk for alcoholism. PMID- 9347090 TI - Regional brain metabolic response to lorazepam in alcoholics during early and late alcohol detoxification. AB - Changes in GABA function have been postulated to be involved in alcohol tolerance, withdrawal and addiction. In this study we measured regional brain metabolic responses to lorazepam, to indirectly assess GABA function (benzodiazepines facilitate GABAergic neurotransmission), in alcoholics during early and late withdrawal. Brain metabolism was measured using PET and 2-deoxy 2[18F]fluoro-D-glucose after placebo (baseline) and after lorazepam (30 micrograms/kg intravenously) in 10 alcoholics and 16 controls. In the alcoholics evaluations were performed 2 to 3 weeks after detoxification and were repeated 6 to 8 weeks later. Controls were also evaluated twice at a 6 to 8 weeks interval. While during the initial evaluation metabolism was significantly lower for most brain regions in the alcoholics than in controls in the repeated evaluation the only significant differences were in cingulate and orbitofrontal cortex. Lorazepam-induced decrements in metabolism did not change with protracted alcohol withdrawal and the magnitude of these changes were similar in controls and alcoholics except for a trend towards a blunted response to lorazepam in orbitofrontal cortex in alcoholics during the second evaluation. Abnormalities in orbitofrontal cortex and cingulate gyrus in alcoholics are unlikely to be due to withdrawal since they persist 8 to 11 weeks after detoxification. The fact that there was only a trend of significance for an abnormal response to lorazepam in orbitofrontal cortex indicates that mechanisms other than GABA are involved in the brain metabolic abnormalities observed in alcoholic subjects. PMID- 9347091 TI - Endocrine and hemodynamic effects of stress versus systemic CRF in alcoholics during early and medium term abstinence. AB - In alcoholics, disturbances of the autonomic nervous system as well as of the hypothalamic-pituitary-adrenal axis (HPA) are known. However, these two systems have never been analyzed, under stimulated conditions, in parallel in the same patients. Moreover, studies using intravenous (i.v.) corticotropin releasing factor (CRF) to assess neuroendocrine function bypass the hypothalamic component of the HPA axis. Therefore, i.v. human (h) CRF (pituitary stimulation/exogenous CRF) and a multifaceted stress test (hypothalamic activation/endogenous CRF) were compared with respect to their effects on hemodynamics as well as plasma norepinephrine (NE), epinephrine (E), ACTH, and cortisol in abstinent alcoholics (n = 11) versus healthy men (n = 10). Each stimulus was tested twice, 12 weeks apart, in two separate experimental blocks (I and II). Alcoholics entered block 18 days after the last ethanol ingestion and were controlled for abstinence up to block II. hCRF caused a fall in mean arterial pressure (MAP), most pronounced in alcoholics, particularly in block II. In contrast, stress testing raised MAP in both groups and blocks. A sustained increase in ACTH, cortisol, and NE occurred after hCRF, although the ACTH response in alcoholics was blunted in both blocks. Stress testing elevated NE in both groups and blocks, while raising plasma ACTH and cortisol during block I only in controls. However, unlike the persistently blunted ACTH response to i.v. CRF, a normalization of the stress-induced ACTH output occurred in alcoholics after 12 weeks of abstinence. During block I, basal E levels were elevated in alcoholics whereas NE levels tended to be lower than in controls, resulting in a significantly decreased NE/E ratio that returned to near control values in block II. Neither CRF nor stress had any effect on circulating E in either group or block. To conclude: (1) Normalization of the ACTH response to stress, but not to i.v. CRF, after 12 weeks of abstinence, suggests that other ACTH secretagogues may be compensating for CRF dysfunction in alcoholics. (2) Despite the dramatically lowered plasma NE/E ratio in alcoholics, the NE response to stimuli was unaffected. (3) The exaggerated hypotensive reaction and blunted ACTH response to i.v. CRF may reveal a long-term dissociative dysregulation of CRF actions in alcoholics. PMID- 9347092 TI - Behavioral effects and pharmacokinetics of low-dose intravenous alcohol in humans. AB - Alcohol has physiological effects on the human central nervous system at blood alcohol concentrations (BACs) as low as 9 mg/dl. It is unknown, however, if humans can perceive the effects of such low doses of alcohol. Furthermore, low BACs can be difficult to measure. The purpose of this experiment was to: (1) assess the ability of humans to perceive subjective effects of low BACs; (2) measure behavioral effects of low BACs on a psychomotor performance task; and (3) test the sensitivity and accuracy of the transdermal alcohol sensor (TAS) for measuring low BACs from skin. Five men and seven women were administered single blind intravenous infusions of ethyl alcohol in 5% dextrose/water to achieve peak BACs of 0, 10, 20, and 40 mg/dl. Subjective intoxication scales and a computer administered continuous performance task (CPT) were used to assess alcohol effects. BACs were estimated from skin, blood, and breath. The only alcohol induced sensation significantly increased during the alcohol infusions was anesthesia measured by the Alcohol Sensation Scale on the descending limb of the BAC curve. The subjective positive-reinforcing stimulant and mood effects of alcohol were not reported until subjects were administered the 40 mg/dl alcohol infusion. Other measures of subjective intoxication and sedation, and the CPT were unaffected by the alcohol infusions. The TAS provided a noninvasive method for estimating BACs that was comparable with estimates obtained from blood and breath, although delayed in time. PMID- 9347093 TI - Violence, suicidality, and alcohol/drug use involvement in adolescent females with a psychoactive substance use disorder and controls. AB - This study had three aims: (1) to determine the relations between behavioral dysregulation, negative affectivity, and familial impairment with violence and suicidality (i.e., severity of ideation and attempts) in a sample of adolescent females with a psychoactive substance use disorder and controls; (2) to determine whether these relations are mediated by internalizing (depression/anxiety) and externalizing (nonviolent antisocial behavior) symptomatology; and (3) to determine whether severity of alcohol/drug use involvement moderates the relations between the mediating variables with violence and suicidality. Multiple behavioral, psychiatric interview, and self-report measures were used to index behavioral dysregulation, negative affectivity, familial impairment, internalizing and externalizing symptomatology, and violence and suicidality in one hundred sixty-one 14- to 18-year-old adolescent females with a psychoactive substance use disorder and in 80 controls. Structural equation modeling was used to determine the proposed relations. Results indicated that behavioral dysregulation, negative affectivity, and familial impairment were related to violence, whereas only familial impairment was related to suicidality. Internalizing symptomatology mediated the relation between familial impairment and suicidality, and was related to violence, whereas externalizing symptomatology mediated the relations between behavioral dysregulation, negative affectivity, and familial impairment with violence. Severity of alcohol/drug use involvement did not moderate the relations between internalizing or externalizing symptomatology with suicidality or violence. Nevertheless, the relation between internalizing symptomatology and suicidality was stronger in females with a greater degree of alcohol/drug use involvement, compared with those with a milder degree of involvement. Therefore, from a prevention standpoint, behavioral dysregulation, negative affectivity, familial impairment, as well as internalizing and externalizing symptoms, may serve as clinical "points of intervention" for altering the development of violence and suicidality in high risk and substance abusing youth. PMID- 9347094 TI - Effect of pyridostigmine on the thyroid-stimulating hormone response to thyrotropin-releasing hormone in abstinent alcoholics. AB - Alcoholism is sometimes associated with a blunted thyroid-stimulating hormone (TSH) response to thyrotropin-releasing hormone (TRH; peak minus baseline < 5 mIU/liter), despite basal TSH and thyroid hormone levels within the normal range. In light of the inhibitory effect of somatostatin on TSH secretion, we examined whether this condition is caused by an increased hypothalamic somatostatinergic tone in alcoholic subjects. To answer this question, 16 euthyroid male alcoholics (aged 38 to 50 years) with normal [n = 8; normal responder alcoholics (NRAs)] or blunted [n = 8; low responder alcoholics (LRAs)] TSH response to TRH were selected in a preliminary TRH test (200 micrograms in an intravenous bolus). In addition, 8 age- and weight-matched normal men were tested with TRH and used as normal controls (NCs). NCs and alcoholic patients showed similar basal serum TSH, free triiodothyronine, and free thyroxine levels. All subjects were tested again with TRH, 60 min after treatment with 180 mg of pyridostigmine orally. According to selection criteria, NCs and NRA groups showed similar TSH responses to TRH, whereas TRH-induced TSH rise was strikingly lower in LRAs than in NCs and the NRA group. Pyridostigmine did not change the basal levels of TSH in any group, whereas it enhanced in a similar manner the TRH-induced TSH rise in the NC and NRA groups. No significant change in the TSH response to TRH was observed in LRA patients after pyridostigmine treatment. These data argue against the possibility that an enhanced somatostatinergic tone is responsible for the blunted TSH response to TRH observed in some alcoholic patients. This phenomenon might be attributed to alcohol-related defects of stimulated pituitary thyrotroph secretory capacity in some individuals, possibly due to genetic vulnerability and/or the toxic effects of prolonged alcohol abuse. PMID- 9347095 TI - Prepulse inhibition of the startle reflex: preliminary study of the effects of a low dose of alcohol in humans. AB - The present study was designed to examine the effects of a low dose of alcohol on prepulse inhibition (PPI) of the startle response and self-report measures of affect. Eighteen subjects participated in a counterbalanced repeated-measures design in which they received a beverage with alcohol during one session and a nonalcohol beverage during a different experimental session. The startle response was probed in two separate 10-min blocks immediately after consumption of the alcohol. Although alcohol significantly suppressed the startle response in general, it did not do so to an extent that compromised detection of PPI. The effects of alcohol on PPI were primarily evident in the first block and were dependent on baseline levels of PPI, such that alcohol resulted in a reduction of PPI in subjects who demonstrated low PPI at baseline and an increase in PPI for subjects with high PPI at baseline. Alcohol also significantly increased self reported stimulation during the first block and increased negative affect during the second block. These findings suggest that baseline PPI may reflect an important individual difference that is predictive of the direction and magnitude of alcohol-induced changes in sensorimotor gating. PMID- 9347096 TI - Disinhibited personality and sensitivity to alcohol reinforcement: independent correlates of drinking behavior in sons of alcoholics. AB - Thirty nonalcoholic young (18 to 30 years) males with extensive multigenerational family histories of male alcoholism and 29 age-matched, family history-negative controls completed a variety of trait personality questionnaires, participated in a competitive stress task (while sober and alcohol-intoxicated), and were assessed for self-report and laboratory drinking behavior. Low academic achievement, disinhibited personality (as measured by the P Scale of the Eysenck Personality Questionnaire), and sensitivity to alcohol reinforcement were significant and powerful independent predictors of self-report (approximate R2 = 0.40, p < 0.0001) and laboratory (approximate R2 = 0.20, p < 0.0001) drinking behavior. There seemed to be some specificity with respect to the facets of drinking behavior accounted for by each independent variable: low academic achievement and sensitivity to alcohol reinforcement were more related to quantity of alcohol consumption and frequency of excessive consumption, whereas psychoticism was more related to self-reported negative consequences with alcohol. A cluster analysis on three identified correlates of drinking behavior indicated that the two experimental groups could be more accurately subdivided into three homogeneous types. Multigenerational family history males were disproportionately represented in two of these groups: one characterized by enhanced sensitivity to alcohol reinforcement and the other characterized by high psychoticism scores and alcohol-related problems. PMID- 9347097 TI - Prodynorphin allelic distribution in Scandinavian chronic alcoholics. AB - Two regions of the human prodynorphin gene, the exon 4-region coding for the opioid peptides and the putative promotor/exon 1-region were analyzed for possible presence of polymorphisms. No polymorphism was detected in the exon 4 region, whereas a GC/AT base-pair exchange was observed 301 base pairs upstream of the exon 1/intron A boundary. This polymorphic marker was examined in Scandinavian chronic alcoholics (n = 70) and control subjects (n = 55). Prodynorphin allelic distributions were not significantly different in alcoholic patients and control subjects. The results suggest that no major influence on alcoholism is exerted through genes associated with this prodynorphin allelic marker. PMID- 9347098 TI - Is carbohydrate-deficient transferrin a specific marker for alcohol abuse? A study in patients with chronic viral hepatitis. AB - Carbohydrate-deficient transferrin, a transferrin isoform, is hailed as a new marker of chronic alcohol abuse, but its specificity is, however, not unequivocally accepted. The aim of the present study was therefore to determine carbohydrate-deficient transferrin levels in patients with chronic hepatitis B and C with or without documented chronic alcohol intake. Carbohydrate-deficient transferrin was measured using a double-antibody radioimmunoassay (CDTect, Pharmacia) in serum samples from 66 patients (45 males and 21 females; mean age: 39 years) with chronic viral hepatitis B (n = 20) or C (n = 46). Diagnosis of the underlying liver disease was established by liver biopsy. Carbohydrate-deficient transferrin levels were raised in 15 patients [23%; hepatitis B (n = 2) and hepatitis C (n = 13)]. In patients with chronic hepatitis B, the carbohydrate deficient transferrin level was raised in two abstainers. In the 46 patients with chronic hepatitis C, 10 (22%) patients with an alcohol consumption of < 60 g/day for the men and 30 g/day for the women had raised carbohydrate-deficient transferrin levels. The overall specificity of carbohydrate-deficient transferrin for chronic alcohol abuse was thus 78%, suggesting an association between elevated carbohydrate-deficient transferrin levels and the presence of chronic viral hepatitis. Carbohydrate-deficient transferrin levels were not correlated with the histological grading or staging of chronic hepatitis B and C, or with biological markers of hepatic synthesis and cellular damage. Thus, an increased carbohydrate-deficient transferrin level may occur in patients with chronic viral hepatitis in the absence of chronic alcohol abuse. This fact should be kept in mind by physicians when using this marker to detect alcohol abuse. PMID- 9347099 TI - Utility of biological markers during outpatient treatment of alcohol-dependent subjects: carbohydrate-deficient transferrin responds to moderate changes in alcohol consumption. AB - A group of 25 alcohol-dependent subjects in outpatient treatment were monitored for a period of 4 weeks. They were weekly interviewed for their alcohol consumption and their serum levels of carbohydrate-deficient transferrin (CDT) and gamma-glutamyltransferase (GT) were analyzed. The majority of the patients reported an excessive and fairly constant alcohol intake during the observation period. When selecting those patients that reported periods of 1 or 2 weeks with moderate changes in alcohol consumption, corresponding changes in CDT were demonstrated. Thus, of 14 patients reporting an increased alcohol consumption for 2 weeks (mean values increased from 57 to 101 g/day), 11 showed an increase in CDT at the end of the period. The mean CDT value of all 14 increased from 5.5 to 6.7% (p < 0.05). Slight, but not significant, increases were noted in GT, indicating that CDT is more sensitive than GT in detecting increased alcohol consumption. Furthermore, of 17 patients that reported decreased alcohol consumption for one or several weeks, 14 showed decreased CDT and GT values. The mean values of all 17 were reduced from 5.1% to 4.5% (CDT) and from 126 units/liter to 97 units/liter (GT) (p < 0.05 for both parameters). The results indicate that CDT responds to moderate changes in alcohol consumption in alcohol dependent patients and may thus be useful as a corrective tool to self-reports of alcohol consumption during outpatient treatments. PMID- 9347100 TI - Absence of hepatitis G virus infection in patients with alcoholic cirrhosis. PMID- 9347101 TI - Thalidomide recommended for approval under tight restrictions. PMID- 9347102 TI - National goals for child immunization being met, but shortfalls remain at state and local levels. PMID- 9347103 TI - Tracking and reporting drug expenditures. PMID- 9347105 TI - Standardized antiemetic regimens. PMID- 9347104 TI - Haunting medicines. PMID- 9347106 TI - Soft tissue limitations in orthodontics: treatment planning guidelines. AB - Orthodontists have traditionally viewed structural discrepancies as the major limitation of treatment. In reality, it is the soft tissues that more closely determine therapeutic modifiability. The boundaries of dental compensation for an underlying jaw discrepancy are established by pressures exerted on the teeth by the lips, cheeks, and tongue; limitations of the periodontal attachment; neuromuscular influences on mandibular position; and the contours of the soft tissue facial mask. The ability of the soft tissues to adapt to changes in tooth jaw relationships are far narrower than the anatomic limits in correcting occlusal relationships. The tolerances for soft tissue adaptation from equilibrium, periodontal, and facial balance standpoints are in the range of 2 to 3 mm for expansion of the mandibular arch and even less for changes in condylar position. Thus, analysis of the soft tissues is the critical step in orthodontic decision making, and this can only be accomplished through physical examination of the patient. Although quantitative measurements cannot be rigorously applied, guidelines for soft tissue assessment, with particular emphasis on facial esthetics, are proposed. From this perspective, a contemporary philosophy of orthodontic practice is offered, with general indications and contraindications for nonextraction, extraction, and surgical treatment. PMID- 9347107 TI - Predicting soft tissue changes in mandibular advancement surgery: a comparison of two video imaging systems. AB - The purpose of this study was to evaluate the accuracy of two video imaging systems, Prescription Portrait and Orthognathic Treatment Planner, in predicting the soft tissue profiles of 39 patients who underwent mandibular advancement surgery. Presurgical cephalograms and profile photographs were entered into a computer. Computerized cephalometric line and video image predictions were generated and compared with the actual postsurgical results. The results indicate that both programs were equally accurate clinically in their line drawing and video image predictions. In the line drawings, clinically acceptable accuracy was shown in approximately 80% of the upper lip and chin predictions and in less than 50% of the lower lip predictions. The video images produced by both programs received fair to good ratings from a panel of professional and lay judges. Orthodontists and surgeons rated all aspects of the images similarly, while lay people were most critical of the chin and submental areas and least critical in their overall evaluation. PMID- 9347108 TI - Predicting soft tissue changes in maxillary impaction surgery: a comparison of two video imaging systems. AB - The purpose of this retrospective study was to investigate the accuracy of two video imaging systems, Orthognathic Treatment Planner (OTP) and Prescription Portrait (Portrait), in predicting soft tissue profile changes after maxillary impaction surgery. Computer-generated line drawing predictions were compared with actual postsurgical profiles. Neither program was very accurate with vertical measures and lower lip contour. Portrait was more accurate at pronasale, inferior labial sulcus, and pogonion in the y-axis direction (P < 0.05). Video image predictions produced from the presurgical photographs were rated by orthodontists, surgeons, and lay people, who compared the predictions with the actual postsurgical photographs using a visual analog scale. Portrait's prediction images were scored higher than OTP's for five of eight areas. Orthodontists were most critical of the lips and the overall appearance. Lay people were most critical of the chin and submental areas. PMID- 9347110 TI - A comparison of sonically derived and traditional cephalometric values. AB - The roentgenocephalometric technique is the standard used by orthodontists to assess skeletal, dental, and soft-tissue relationships. However, this technique exposes patients to radiation, preventing orthodontists from taking frequent cephalograms to assess growth and to monitor treatment. Recently, the Dolphin Imaging Company developed the DigiGraph, a nonradiographic cephalometric method that uses sound waves and mathematical algorithms, and consequently does not expose patients to radiation. But the DigiGraph's accuracy as a cephalometric alternative has not been adequately investigated. The purpose of this study was to compare the values obtained by traditional cephalometrics with those obtained by the DigiGraph technique for 30 well-known measurements, and then to assess the repeatability (intraobserver comparison) and reproducibility (interobserver comparison) for both techniques. Eighteen of the 30 measurements had mean differences that were statistically significant (p > .0067). Regression plots generally illustrate low correlations for the measurements, although Ricketts' esthetic line (upper and lower lip) and Steiner's soft-tissue convexity reveal strong linear relationships between the two methods. Additionally, the radiographically generated measurements showed greater repeatability and reproducibility. PMID- 9347109 TI - The accuracy of video imaging for mixed dentition and adolescent treatment. AB - The purpose of this study was to evaluate the accuracy of computerized video imaging in predicting the soft tissue outcome of growth modification treatment for skeletal Class II malocclusions. Pretreatment and posttreatment cephalometric and facial photographic records of 22 mixed dentition (8 to 10 years old) and 20 adolescent (12 to 14 years old) patients were digitized, and the known outcomes were compared with computer-generated VTOs and video images. The predicted video images were found to be reasonably accurate for the mixed dentition group, but unacceptable for the adolescent group. When graded by a panel of judges, orthodontists were far more critical of the findings than their lay counterparts. These results emphasize the potential of video imaging as a communication medium, rather than as a diagnostic tool for growing patients. PMID- 9347111 TI - Soft tissue profile changes in late adolescent males. AB - The purpose of this study was to document soft tissue profile changes in late adolescent skeletal Class I males from 14 to 20 years of age and to compare these changes with those of the underlying hard tissues. Using serial lateral cephalograms from a sample of 33 untreated Class I adolescent males, 26 soft and hard tissue parameters were assessed at ages 14 to 16, 16 to 18, and 18 to 20 years. The concept that differential facial growth occurs from nasion to pogonion was substantiated by these data. The hard tissue chin moved forward more than A point, which in turn moved forward more than nasion, resulting in the hard and soft tissue profile being flattened or reduced in convexity. Horizontal soft tissue thickening of 1.0 mm overlying the hard tissue surfaces from midface to chin was observed at 14-16 years. Continued change of the soft tissue profile from 16 to 20 was thus the result of underlying skeletal growth. Nasal tip increased significantly over all age periods, and underwent the largest growth change of all measurements assessed (approximately 8.0 mm). This growth increase declined by approximately one-half over each successive age period. Although variable, continued soft tissue movements throughout the 14- to 20-year age period affect treatment planning, maintenance of the posttreatment profile, and posttreatment occlusal retention requirements. PMID- 9347112 TI - Mesh diagram analysis: developing a norm for Puerto Rican Americans. AB - Cephalometric radiography is an important diagnostic aid in orthodontics. Mesh analysis is a proportionate cephalometric method of graphically assessing disharmonies of the craniofacial complex. Original norms for this analysis were created from a white, European American sample. Norms for black Americans of African descent were developed in another study. The purposes of this investigation were: (1) to develop a standard mesh diagram from a Puerto Rican American population; (2) to compare the diagram with previously established data from the white sample; (3) to develop linear and angular means for the Legan, Burstone, Ricketts, DiPaolo, and Steiner analyses for a Puerto Rican American population; and (4) to assess the use of a panel for selecting esthetically pleasing faces. The subjects in the study had no previous orthodontic treatment, had Class I occlusion with 6 mm or less of crowding per dental arch, and had two parents and two sets of grandparents who were were born in Puerto Rico. Sixty nine patients met the study criteria, and 50 of those patients (20 males and 30 females) were selected as having esthetically pleasing faces by the panel. Male and female norm diagrams were created and these were compared with those developed previously. Linear and angular measurements were also compared. Significant differences between the ethnic groups were found in the dentoalveolar region. Similarities were noticed in the upper face height and anterior cranial base length. The panel selection results showed no agreement within the sexes, occupations, or ethnic groups. PMID- 9347114 TI - Cephalometry needs innovation, not renovation. AB - Units of length, degree, and area are used when measuring cephalograms. In particular, the measurement of angles is a conventional method of quantifying shape. Because angles do not provide information about direction, there is no way to tell how and where one part of the facial structure has moved with respect to the rest. A new landmark data system using x- and y-coordinates is proposed, and some of its advantages over conventional methods are explained. PMID- 9347113 TI - A retrospective comparison of frontal facial dimensions in alveolar-bone-grafted and nongrafted unilateral cleft lip and palate patients. AB - This retrospective study was undertaken to describe and compare frontal craniofacial dimensions in alveolar-bone-grafted and nongrafted complete unilateral cleft lip and palate (CUCLP) patients and in noncleft subjects with normal occlusions and good facial balance. Clinical data were obtained from the files of the Hospital for Sick Children, Toronto. Patients were eligible for inclusion if they had posteroanterior cephalograms (PA) taken at adulthood and no congenital anomalies other than CUCLP. A total of 86 adult Caucasian CULCP patients were studied, including 58 who had not received grafts, 28 who had received secondary alveolar bone grafts, and, for comparison, 60 noncleft Caucasian adults. The PA cephalometric radiographs were traced, digitized, and measured. Analysis of variance (ANOVA) was used to test for among-groups differences in the means of the ratios, proportions, and angular measures. Tukey Kramer HSD procedure was used to conduct post-hoc pairwise comparisons following significant (p < or = 0.05) F-ratios from ANOVA. Sexual dimorphism was a common finding, with males demonstrating greater facial width. Despite primary surgical repairs, the anterior nasal spine in the nongrafted CUCLP patients was deviated to the noncleft side, and the alar base was depressed on the cleft side. The maxillary incisors close to the cleft site were irregularly inclined, and this irregularity was more severe in the nongrafted CUCLP patients. The long-term effects of secondary alveolar bone grafting on transverse craniofacial growth appears to be minimal and limited to the immediate area of the cleft. PMID- 9347116 TI - Adherence of oral streptococci to an immobilized antimicrobial agent. AB - An antimicrobial agent, 3-(trimethoxysilyl)-propyldimethyloctadecyl ammonium chloride, was immobilized on silica. Interaction between the material (termed) OAIS) and various oral bacterial species were then studied. Seven species of Streptococcus and two Actinomyces were investigated for their ability to adhere to this biomaterial. Cell-surface hydrophobicity and zeta-potential were examined as well. Analysis of extracted hydrophobic proteins which adhered to OAIS revealed that the adherence of these micro-organisms was closely related to the hydrophobicity of their cell surfaces. The results of zeta-potential assays indicated that negative charge on the cell surface inhibited adherence to OAIS. Gel electrophoresis revealed that OAIS could absorb cell-surface hydrophobic proteins from all bacterial species tested. Preadsorption of hydrophobic components on the cell surface inhibited adherence of the Strep. mutans strain to OAIS in a dose-dependent manner. The results indicate that OAIS adsorption of these oral bacteria was dependent on the degree of hydrophobicity of their surfaces. A major component of this adherence was hydrophobic cell-surface proteins. PMID- 9347115 TI - Characterization of human sublingual-gland protein kinase by phosphorylation of a peptide related to secreted proteins. AB - Phosphoproteins in human saliva include proline-rich proteins, statherins, histatin 1 and cystatin SA-III. The presence of phosphate in these proteins is necessary for various functions in the mouth including calcium binding, inhibition of precipitation of calcium phosphate, inhibition of growth of hydroxyapatite crystals and adherence to hydroxyapatite. To elucidate the process of phosphorylation of these proteins, the phosphorylation of a peptide (APRP8) with an amino acid sequence identical to one of the phosphorylated sites in acidic proline-rich proteins by a kinase from the human sublingual gland was investigated. The kinase, which was highly labile, was purified 58-fold by fractionation of sublingual gland homogenate and gel filtration, but the enzyme was inactivated when further purification by chromatographic techniques commonly used for protein kinases was attempted. To compare the enzyme with other kinases, and to obtain information that could be used in its further purification, a characterization was undertaken. The enzyme required 10 mM Mg2+ for optimum activity, it had a KM of 0.09 mM for ATP and the KM for the peptide substrate APRP8 was 0.42 mM. It was not activated by cAMP or calmodulin, characteristics that are shared with casein kinases and mammary gland kinase. The sublingual kinase as well as casein kinase 2 were inhibited by heparin, but in other respects the two kinases had different properties. While casein kinase 2 is activated by polylysine and has optimal activity in 150 mM KCl, sublingual kinase was inhibited by polylysine and the addition of KCl. Moreover, casein kinase 2 can utilize both ATP and GTP as phosphoryl donors, but GTP was not a substrate for sublingual kinase. The sublingual kinase shared a substrate recognition sequence with mammary gland kinase, but, unlike that kinase, it could not utilize Ca2+ instead of Mg2+. While the sublingual kinase thus shared some properties with both casein kinase 2 and mammary gland kinase, distinct differences were also seen and the relationship to these enzymes remains to be determined. The characterization of the sublingual kinase will be useful in its further purification. PMID- 9347117 TI - A calcium channel in human submandibular duct cell line, HSG cells, not regulated by P2U purinergic receptor-mediated intracellular calcium mobilization. AB - Signal transduction via P2 purinergic receptors was investigated in HSG cells, a continuous cell line originally derived from an irradiated human salivary gland. Ligand specificity for nucleotide receptors in HSG cells was investigated with various nucleotides and their analogues. Inositol 1,4,5-trisphosphate (IP3) production was significantly increased by ATP, UTP and ATP gamma S. The ligand specificity of this effect agreed well with that of the P2U purinergic receptor. On the other hand, 45Ca2+ influx was stimulated by ATP, UTP > ATP gamma S, ADP, UDP > ADP beta S > AMPPNP, GTP, TTP > CTP, GDP, TDP, AMPPCP, AMPCPP. This ligand specificity of 45Ca2+ influx was much broader than IP3 production. Also pertussis and cholera toxin had no effect on both IP3 production and 45Ca2+ influx by ATP or UTP. 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate (Bz-ATP) stimulates 45Ca2+ influx more effectively than IP3 formation. A 53-kDa membrane protein was photolabelled with [alpha-32P]Bz-ATP. This 53-kDa protein is a putative P2 purinergic receptor. In particular, the labelling was inhibited by a ligand profile that corresponded to that for 45Ca2+ influx. These findings suggest that nucleotides stimulate 45Ca2+ influx and IP3 formation by separate pathways via pertussis and cholera toxin-insensitive G proteins. Thus, in HSG cells, IP3 formation is coupled to the P2U subclass, while 45Ca2+ influx is coupled to another subclass, such as P2X, that regulates calcium channels. PMID- 9347118 TI - Detection of human herpesvirus 7 in salivary glands. AB - The prevalence and cellular distribution of human herpesvirus 7 (HHV-7) in archival labial salivary glands was analysed for virus-specific DNA sequences by polymerase chain reaction (PCR) and in situ hybridization signals. In addition, the cellular expression of HHV-7-encoded protein was detected by immunohistochemical staining with a virus-specific monoclonal antibody. Eleven of 20 samples were positive for the HHV-7 DNA sequence by PCR. Eighteen of 20 tissues analysed by in situ hybridization showed signals in ductal, serous and mucous cells. Some nuclei of these cells and also the myoepithelial population were positive. In immunolocalization studies, all 20 salivary glands consistently showed HHV-7-expressed protein in the cytoplasm of ductal cuboidal and columnar cells. The protein was also found in the cytoplasm of mucous and serous acinar cells that were immunopositive for HHV-7. The observations are consistent with the suggestion that the labial salivary gland is a site for virus replication, potential persistence and a source of infective HHV-7 in saliva. PMID- 9347119 TI - Calcium-calmodulin-stimulated phosphorylation of rat parotid secretion granule proteins. AB - In studies designed to determine the mechanism by which Ca++ and calmodulin stimulate the fusion of parotid secretion granules with plasma membrane vesicles, the hypothesis tested was that Ca++ and calmodulin act by stimulating protein phosphorylation. It was earlier found that Ca++ and calmodulin, but neither alone, stimulated the phosphorylation of four secretion granule proteins with molecular masses of 64, 58, 55 and 31 kDa, and decreased the degree of phosphorylation of a 36-kDa protein. Further studies have shown that in the presence of an optimal concentration of calmodulin (2.4 microM), half-maximal activation of phosphorylation of the four proteins occurred at approx. 8 microM Ca++, and at a maximally effective Ca++ concentration (10(-4) M), half-maximal stimulation occurred at calmodulin concentrations between 0.13 and 1.1 microM for the different proteins. The studies now described also demonstrate that the need for calmodulin for stimulating the phosphorylation, but not the dephosphorylation, is specific; two other Ca(++)-binding proteins, parvalbumin and troponin, could not replace calmodulin in stimulating phosphorylation of the four secretion granule proteins, but either one could substitute for calmodulin in stimulating dephosphorylation of the 36-kDa protein. Additionally, the phosphorylated proteins appear to be located on the granule surface. When secretion granules were subjected to mild treatment with a concentration of trypsin that did not lyse the granules, the 31-, 36-, 55-, 58- and 64-kDa proteins were no longer observed. In the presence of optimal concentrations of Ca++ and calmodulin, a dose-dependent inhibition of the phosphorylation of the various proteins by two calmodulin antagonists, trifluoperazine and calmidazolium, was observed; 50% inhibition of phosphorylation of the different proteins was obtained at approx. 20-40 microM trifluoperazine and at about 2.5 3.0 microM calmidazolium. Inhibition of the dephosphorylation of the 36-kDa protein required greater concentrations of trifluoperazine and calmidazolium; 128 microM and 50 microM, respectively. These results are consistent with the hypothesis that the phosphorylation of one or more of the 31-, 55-, 58- and 64 kDa proteins, but not the dephosphorylation of the 36-kDa protein, may be involved in the action of Ca++ and calmodulin in secretion granule-plasma membrane fusion. PMID- 9347120 TI - The craniofacial complex in 47,XYY males. AB - Eight adult, Finnish 47,XYY males were compared with population male and female controls and, in addition, three of them were compared with first-degree male relatives. Linear and angular measurements were made from standardized lateral cephalograms of patients and normal population controls from the "Kvantti" study series. In both comparisons the craniofacial dimensions in 47,XYY males were larger than those in population male and female controls. Their craniofacial proportions and plane angles were similar to those of normal men except for a larger lower facial height with posterior rotation of the mandible and a tendency to bimaxillary protrusion, a longer cranial base and a lesser cranial-base angle. Thus the supernumerary Y chromosomal gene(s) in 47,XYY males may result in larger craniofacial dimensions than in normal males, without substantial effects on dimensional ratios and plane angles. This general metric pattern is similar to that observed in relation to many adult body and head dimensions, and the dental arches and tooth crowns, of 47,XYY males. The foramen magnum in 47,XYY males was smaller in the sagittal plane than that of normal males and females. PMID- 9347121 TI - Extreme bilateral molar rotation in Monodelphis domestica (Marsupialia: Didelphidae). AB - Rotation of a tooth around an axis perpendicular to the occlusal plane through angles approaching 180 degrees is a rare anomaly found in the mammalian dentition. A specimen of Monodelphis domestica was found to show such extreme rotation of both lower last molars, with consequent disruption of normal occlusion and wear. A review of the literature discovered 41 other reported cases of extreme rotation, from four different orders of mammals. The distribution of extreme rotation within the dentition can be summarized as follows. It is found only in isolated teeth or in contralateral pairs of teeth. Bilateral rotation is far more common than would be expected based on the chance of the independent occurrence of two rotations. Extreme rotation has a significantly higher frequency in upper rather than lower teeth, in premolars rather than other teeth, and on the left- rather than the right--hand side. The incidence of extreme rotation across mammals was estimated to be approx. 1 in 5850. PMID- 9347123 TI - Exclusion of five trinucleotide repeat (CAG and CCG) expansions in 17 families with schizophrenia. AB - Trinucleotide repeat expansion diseases are characterized by anticipation. Several studies suggested the anticipation in schizophrenia. We studied five novel trinucleotide repeats identified by Li et al in a human brain cDNA library in 100 unrelated control subjects, and 57 subjects in 17 families with schizophrenia. The CTG-B43 and B45d, and CCG-A3 were not polymorphic in any unrelated control subjects or subjects with familial schizophrenia. The CTG-A4 had two alleles and the CTG-B1 three alleles. The two clones were not expanded in subjects with familial schizophrenia or unrelated healthy controls. There was no difference in allele frequency between unrelated control subjects and subjects in the families with schizophrenia. The allele frequencies in unaffected and affected subjects in the families with schizophrenia were not significantly different. There was no difference between the offspring and parental generations of affected subjects in the families with schizophrenia. The results suggest the exclusion of the five trinucleotide repeat expansions in the 17 families with schizophrenia. PMID- 9347122 TI - Lack of enhanced response to repeated d-amphetamine challenge in first-episode psychosis: implications for a sensitization model of psychosis in humans. AB - Behavioral sensitization is the process whereby intermittent stimulant exposure produces a time-dependent, enduring, and progressively more robust behavioral response. This process serves as a model for the development of psychosis, but has been little studied in humans. The authors report results from a double blind, placebo-controlled study of repeated d-amphetamine challenges in 13 patients with first-episode manic or schizophrenic psychosis. Each patient received two daily doses of d-amphetamine (0.25 mg/kg) separated by 48 hours that alternated with two daily doses of matched placebo. Symptoms (activity/energy level, mood, rate and amount of speech, and severity of psychosis) and eye-blink rates were measured hourly for 5 hours following drug administration. In contrast to results from previous work in normal volunteers, none of the measures demonstrated the progressive increase following the second amphetamine dose as compared to the first dose that characterizes sensitization. These results suggest that patients with psychosis are already maximally sensitized, so cannot exhibit progressive behavioral enhancement following repeated stimulant challenges, or that patients with psychosis do not sensitize. PMID- 9347124 TI - Anticipation and imprinting in schizophrenia. AB - Anticipation, i.e., a decrease in the age of onset and/or an increase in the severity of a disease in subsequent generations, and imprinting, i.e., different modes of parental transmission, have been suggested in trinucleotide repeat amplification diseases. The purpose of this study was to determine if anticipation and imprinting are associated with familial schizophrenia. Two generations of 49 schizophrenics from 24 families were studied. Ages of onset, numbers hospitalized, diagnostic subclasses of schizophrenia, amounts of antipsychotics, positive symptoms, negative symptoms, treatment resistance, and clinical course ratings, were compared between the two generations. The age of onset was significantly lower in the offspring generation, although there was no difference in the severity between the two generations. The negative symptom scores and clinical course scores in the offspring generation for paternal transmission were significantly higher than those for maternal transmission. Our results suggest the presence of imprinting and anticipation in schizophrenia. PMID- 9347125 TI - Anterior medial temporal lobe volumes in polydipsic schizophrenic patients with and without hypo-osmolemia: a pilot study. AB - The volume of certain brain structures, particularly those in the anterior medial temporal lobe, may be reduced to a greater extent in hyponatremic/hypo-osmolemic polydipsic schizophrenics than other schizophrenic patients. To explore if volume reduction is specific to this particular brain region, and if it is fundamentally associated with polydipsia, we imaged the anterior hippocampi/subicula and adjacent temporal lobes in normal males (n = 10) and polydipsic schizophrenic patients with (n = 7) and without (n = 6) hypo-osmolemia. Bilateral hippocampal/subicular (p < .01), but not temporal lobe (p < .30), volumes were diminished in the hypo-osmolemics relative to the other two groups, who resembled each other on these measures. No recognized or putative factor could explain these findings. Thus anterior medial temporal lobe structures appear to be preferentially diminished in hypo-osmolemic polydipsic schizophrenics. Additional studies are needed to more precisely define these anatomic differences and their relationship to these patients' impaired water excretion and severe mental illness. PMID- 9347126 TI - Sensitization with clozapine: beyond the dopamine hypothesis. AB - Clozapine elicits dose-dependent myoclonic jerks in partially restrained rats and induces paroxysmal electroencephalographic changes, myoclonus, and convulsive seizures in a small but significant percentage of patients. With the hypothesis that the central excitatory effects of clozapine may relate to the unique therapeutic activity of this agent, rats were administered repeated alternate day or weekly very low dose (1 mg/kg) injections of clozapine in an attempt to induce the central excitatory effect through sensitization or kindling. Although initial administrations of this dose elicited no motor response or other behavioral change, repeated administration of the same low dose on either the alternate-day or weekly schedule caused increasing numbers of myoclonic seizure-like jerks (MJs) reaching 75-110 MJs/hour by the sixth clozapine injection. Clozapine sensitized animals exhibited a significantly different pattern of early gene expression in two subcortical sites compared with vehicle-treated controls. These findings may have importance for the treatment of psychosis. PMID- 9347127 TI - Eye tracking disorder in schizophrenia is characterized by specific ocular motor defects and is associated with the deficit syndrome. AB - The objective was to determine the relationships between eye tracking disorder (ETD) in schizophrenia, specific ocular motor measures, and the deficit syndrome. Twenty-five normal comparison subjects and 53 schizophrenic patients had eye movements tested with infrared oculography using a sinusoidal target. Patients were assessed with the Schedule for the Deficit Syndrome. For the patients, the distribution of position root mean square error (a global measure of pursuit) was best fit by a mixture of two normal distributions. This information was used to divide the patients into two subgroups, those with and those without ETD. ETD was almost completely accounted for by several specific ocular motor measures and was significantly associated with the deficit syndrome. The finding that ETD was almost completely accounted for by specific measures bridges a gap of interpretation in this field. ETD and the deficit syndrome of schizophrenia may share a common pathophysiology of cerebral cortical-subcortical circuits. PMID- 9347128 TI - Relationships between neuropsychological and oculomotor measures in schizophrenia patients and normal controls. AB - Establishing the relationship between oculomotor and neuropsychological impairments might facilitate a more coherent description of schizophrenia associated neurocognitive deficits. Therefore, we assessed several aspects of neuropsychological and oculomotor function in 25 medicated schizophrenia patients and 24 age-matched controls. Neuropsychological tasks included the Wisconsin Cart Sort Test (WCST), the Trail Making Test (TMT), the Rey Auditory Verbal Learning Test, and finger tapping speed. Oculomotor functions assessed included smooth pursuit, initiation of smooth pursuit, predictive pursuit, fixation, visually guided saccades, remembered saccades, and antisaccades. Among the schizophrenia patients, predictive pursuit performance correlated significantly with finger tapping (dominant hand), TMT (both parts), and one WCST measure (categories completed). The only other significant correlation among the schizophrenia patients was between antisaccade performance and part A of the TMT. Perseverative errors during the WCST and antisaccade performance were the only measures significantly correlated among the normals. Closely related neurocognitive deficits may be responsible for impairments in TMT, WCST, predictive pursuit, and antisaccade performance in schizophrenia. PMID- 9347130 TI - Association between striatal reduction and poor Wisconsin card sorting test performance in patients with schizophrenia. AB - Several findings support the hypothesis that the striatum is implicated in executive functions and in the modulation of goal-directed behavior, and could play a key role in the pathophysiology and in the production of symptoms and signs in schizophrenia. We have studied the relationship between the objective measures of the striatal structures, as evaluated by magnetic resonance imaging (MRI), and the Wisconsin Card Sorting Test (WCST) performance in a schizophrenic sample. Thirty-five schizophrenic patients underwent MRI scans of striatal structure and neuropsychological evaluation of executive functions by WCST. Poor WCST performers had a reduction of the left caudate nucleus and putamen, and right total striatum when compared to 24 healthy controls. Significant correlation coefficients were also observed between neuropsychological indexes and left striatal measures. The findings suggest the existence of a relationship between abnormalities of striatal structure and abnormal executive-type or organizational cognitive functions. PMID- 9347129 TI - Factor structure of symptoms in functional psychoses. AB - Global ratings from the Scale for the Assessment of Positive Symptoms and Scale for the Assessment of Negative Symptoms were subjected to principal-component analysis (PCA) in 80 schizophrenia patients, 76 patients with schizophreniform disorder, 80 patients with schizoaffective and mood disorders, and 78 patients with delusional, brief reactive, and atypical psychoses. The resulting factors were correlated with depressive, manic, and catatonic syndromes, and subjected to a multivariate analysis of variance across DSM-III-R diagnoses. PCAs revealed that psychosis, disorganization, and negative factors were also present in each of the nonschizophrenic groups. The disorganization factor tended to be related to the manic syndrome, and the negative factor to depressive and catatonic syndromes. Overall, the three factors had little diagnostic relevance in functional psychoses, although the negative factor was relatively more characteristic of schizophrenia. The data suggest that positive, negative, and disorganization factors are not specific to schizophrenia; this is consistent with a dimensional view of psychopathology in functional psychoses. PMID- 9347131 TI - Relationship between 3-methoxy-4-hydroxyphenylglycol and homovanillic acid in saliva and plasma of healthy volunteers. AB - Plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) and homovanillic acid (HVA) levels may reflect changes in central noradrenergic and dopaminergic activity, respectively. The relationship between MHPG and HVA in saliva and plasma was investigated to evaluate the utility of salivary metabolite measurement as a relatively noninvasive and useful alternative to plasma analysis. MHPG and HVA in saliva and plasma, collected concurrently, from 12 healthy volunteers, were measured by high-performance liquid chromatography. Concentration of free MHPG in saliva correlated significantly with plasma free MHPG. Salivary free MHPG was significantly higher than in plasma. Enzymatic hydrolysis of conjugated MHPG corroborated other work that plasma free MHPG, MHPG-glucuronide, and MHPG-sulfate were in roughly equal proportions. Unpredictably, in saliva, free MHPG was greater than 80% of the total. Salivary and plasma free HVA concentrations also correlated significantly, but salivary HVA levels were significantly lower than in plasma. Conjugated HVA was consistently less than 10% of total both in saliva and plasma. These findings suggest that salivary MHPG and HVA can reflect plasma metabolite levels. Although local factors may influence their formation and concentration in saliva, large changes in plasma free MHPG or HVA could be reflected by parallel changes in saliva. PMID- 9347132 TI - Specificity of the pyridostigmine/growth hormone challenge in the diagnosis of depression. AB - Acetylcholine is a neurotransmitter that has been implicated in the pathophysiology of major depression. This is supported by the enhanced growth hormone (GH) release in response to pyridostigmine (PYD) challenge in depressed subjects relative to healthy comparison subjects. The aim of this study is to examine the specificity of the PYD/GH challenge in the diagnosis of depression. Pyridostigmine 120 mg orally, was administered to a total of 116 physically healthy subjects. Growth hormone responses were studied in 38 patients with (DSM III-R) major depression, 13 subjects with panic disorder, 9 subjects with schizophrenia, 10 recently detoxified alcoholics, and a comparison group of 46 healthy volunteers. Mean delta GH (the difference between basal and maximal GH following PYD) was significantly greater than comparison subjects in patients with major depression. Responses observed in patients with schizophrenia and alcohol dependence syndrome did not differ from the comparison group. Those patients with panic disorder and a high Hamilton depression score had an enhanced delta GH. The sensitivity of the PYD/GH test was 63% for major depression. These results indicate that the PYD/GH test may help distinguish depression from schizophrenia, alcohol-dependence syndrome, or panic disorder with a low Hamilton depression score. PMID- 9347133 TI - Is alexithymia the emotional equivalent of blindsight? PMID- 9347134 TI - A single administration of dehydroepiandrosterone does not enhance memory performance in young healthy adults, but immediately reduces cortisol levels. PMID- 9347135 TI - Salivary cortisol in Operation Desert Storm returnees. PMID- 9347136 TI - Acute treatment of bipolar depression with gabapentin. PMID- 9347137 TI - Event-related potentials in workers with ongoing occupational exposure. PMID- 9347138 TI - Shackling of broilers: effects on stress responses and breast meat quality. AB - 1. Experiments were conducted to study the welfare and meat quality effects of shackling. In experiment 1, broilers with or without leg problems were shackled (S) for 4 min on a moving line and blood sampled; or handled (H), returned to the crate and sampled after 4 min; or sampled immediately after removal from the crate (control, C). 2. Plasma corticosterone (CORT) concentrations, as measured by radioimmunoassay, were highest in S and lowest in C, while the H group was intermediate. Leg problems had no effect on CORT. 3. In experiment 2, tonic immobility (TI) was induced in broilers after 2 min inverted handling to determine fear responses. One week later, the birds were fasted, transported and then shackled on a moving shackle line for 0, 1, 3 or 4 min, then unshackled and blood sampled. Wing flapping during shackling was also quantified. 4. Shackling time did not influence CORT concentrations. There was a negative correlation (r = -0.714) between CORT and wing flapping duration in the 1 min shackling treatment. There was no relationship between TI and wing flapping or CORT. 5. In experiment 3, broilers were exposed to two food withdrawal (FW) times (food withdrawn overnight or during crating only) and held for 4 h prior to processing, shackled (0, 2 or 4 min shackling time, ST), and then killed by exsanguination. Blood samples were collected during the neck-cut. Pectoralis superficialis and Supracoracoideus samples were either collected after 15 min and individually quick frozen (IQF) in liquid nitrogen or collected at 4 h post mortem from carcases chilled on slush ice (COI). 6. CORT increased significantly with increased ST. There was a FW x ST interaction effect on the initial pH of fillets. ST influenced the b*, chroma and Hue values of the COI fillets. FW influenced the L* and Hue values of both IQF and COI fillets as well as the a* value of the COI fillets. 7. In summary, CORT increased with shackling time when birds were held after transport. FW and ST also influenced the colour of fillets, although it is not clear whether these changes are perceptible to the consumer. The duration of wing flapping during shackling did not appear to be related to fearfulness, although it was influenced by properties of the shackle line. We suggest that there be a maximum time lapse between shackling and stunning or killing of 2 min to minimise stress and meat quality changes. PMID- 9347139 TI - Feather pecking behaviour in White Leghorns, a genetic study. AB - 1. Genetic variables of feather pecking (FP) behaviour in a 1993 commercial pure line of White Leghorns were estimated at the age of 6, 38 and 69 weeks. 2. Heritability estimates of performing FP were 0.05 +/- 0.06, 0.14* +/- 0.07 and 0.38** +/- 0.12 for 6, 38 and 69 weeks respectively for sum of pecks (PECKS) and 0.13* +/- 0.07, 0.13* +/- 0.07 and 0.35** +/- 0.12 for sum of bouts (BOUTS). 3. Heritability estimates of receiving FP were not significantly different from 0 except at 6 weeks (0.15* +/- 0.07 and the average of the 3 age classes (AVG) (0.22** +/- 0.09) for PECKS and at 6 weeks (0.15* +/- 0.07) using BOUTS. 4. Genetic correlations of performing FP among age classes were in general high and significant. This was not the case with receiving FP. 5. Plumage cover at 51 weeks had a negative genetic correlation with performing FP at 69 weeks and AVG, but not with receiving FP. No phenotypic correlations were significant between plumage and FP. 6. Body weight at 51 weeks had a negative genetic correlation with performing FP at AVG. 7. Heritability estimates for performing and receiving FP at 6 weeks correspond to those in the literature. No estimates have previously been reported on feather pecking at 38 weeks or 69 weeks. 8. Selection of birds with no or a very low tendency to perform feather pecking should, on the basis of our results, be feasible. PMID- 9347140 TI - Effect of dietary fatty acids on humoral immune response of turkeys. AB - 1. This study examined the effect of increasing amounts of dietary polyunsaturated fatty acids on the fatty acid composition in serum and antibody production following a standard vaccination programme in growing turkeys. Turkey poults were fed on 5 diets containing 75g/kg added fat made up of different proportions of palm and soyabean oils, and were vaccinated against Newcastle disease, infectious bronchitis and necrotic enteritis according to a standard vaccination programme. Blood samples were taken before and one week after each vaccination. 2. Fatty acid composition in serum reflected the composition of the diets although arachidonic acid concentration was not changed by dietary fatty acid content. Growth, erythrocyte and leukocyte parameters were not affected by the respective diets. 3. Specific antibody production was related quadratically to serum linoleic and total n-6 polyunsaturated fatty acid concentrations. No correlation was found with linolenic or arachidonic acids. 4. It is concluded that dietary fatty acid composition can augment the specific anti-vaccine immune response in turkey poults. PMID- 9347141 TI - Effect of beta-aminoproprionitrile on eggshell formation. AB - 1. Eggshells are bioceramic-biopolymer composites made by a cell-mediated deposition of an extracellular matrix which drives the organisation of the inorganic phase. Ultrastructurally, eggshells are composed of shell membranes, mammillary knobs, palisade, and cuticle. Shell membranes are two nets of type X collagen-containing fibrils. On to these membranes, the mammillary knobs, that is, the crystal nucleation sites, are deposited. Type X collagen is highly cross linked and insoluble. 2. In order to evaluate the role of type X collagen cross linking on eggshell formation, hens were injected with different doses of beta aminoproprionitrile, which specifically interferes with cross-link formation. 3. Changes in egg size and shape were observed. Scanning electron micrographs analysis of these eggs demonstrated marked changes in crystal growth and shell membrane structure and arrangement. A dot-blot analysis, using a monoclonal antibody against chicken type X collagen, shows a dose-dependent increase in shell membrane collagen extractability. 4. It is concluded that the formation of beta-aminoproprionitrile-sensitive cross-links among the type X collagen molecules of the shell membranes play an essential role in normal eggshell formation. PMID- 9347142 TI - Problem of pale soft exudative meat in broiler chickens. AB - 1. The occurrence of the pale, soft exudative (PSE) problem in broiler breast meat was monitored in 700 birds (random samples from 7 flocks). A rapid method of colour measurement was used in the plant to assess the colour of skinless pectoralis muscles at 24 h post mortem. In addition, 10 selected samples from each flock were further evaluated for pH, water holding, cook loss, and texture. 2. The average L* value (lightness) was 47.1 with a variance of 5.38, skewness 0.33 and kurtosis 0.66. Minimum and maximum L* values recorded during the study were 35.3 and 55.5 respectively. 3. Correlations between colour and most of the physical measurements were observed, suggesting that a rapid colour measurement method could be used to recognise PSE meat or identify flocks with a high incidence of PSE. 4. Breast muscle samples with L* > 49 showed poor water holding capacity, which indicated that these samples would be classified as PSE meat. Using this criterion it was shown that the occurrence of PSE ranged from 0% to 28%. However, the precise cut off point for PSE muscle should be determined by each processor depending on final product requirements. PMID- 9347143 TI - Effects of selection, over three and four generations, on meat yield and fatness in Muscovy ducks. AB - 1. Two experiments were undertaken to analyse the effects of selection for lowering carcase fatness and improving meat yield of Muscovy ducks. The control generation N and the selected generations N + 3 and N + 4 of the same heavy line (Grimaud) were reared under similar conditions. 2. We compared growth, carcase characteristics, chemical composition of breasts and plasma concentrations of very low density lipoproteins (VLDL), triglycerides and phospholipids. 3. Selection induced an increase of body weight (+8% to 10% at slaughter age), a decrease of abdominal fat percentage (-10%) and an improvement of breast and thigh plus shank yields (+3% to 7% and +4% respectively). 4. The lipid content of breast meat decreased in the selected ducks (-14% to -20%), particularly phospholipids and triglycerides. Breasts appeared paler and less red which suggested modifications of muscular fibre composition. 5. We found no significant correlations between plasma VLDL, triglyceride and phospholipid concentrations and carcase fatness. It therefore seems difficult to use these variables as selection criteria for lowering carcase fatness of Muscovy ducklings. PMID- 9347145 TI - Preincubation storage of turkey eggs: impact on rate of early embryonic development. AB - 1. The effect of cooling and storage on preincubation development of turkey embryos was examined. 2. Fresh laid eggs were stored at 15 degrees C for 3, 7, or 14 d (group 1) or held initially at room temperature (21 degrees C) for 6 to 9 h and subsequently stored at 15 degrees C for 3, 7, or 14 d (group 2). 3. Most embryos in eggs from both groups progressed developmentally to Stage-8, just prior to onset of hypoblast formation. There were no significant differences in embryo development either within or between groups. 4. It is concluded that under the conditions of this study, egg cooling and storage augment development of the embryo, albeit only slightly. PMID- 9347146 TI - Carcase and component yields of rheas. AB - 1. Three Greater Rheas (Rhea americana) and 5 Lesser Rheas (Pterocnemia pennata) were slaughtered, using the procedures conventional for ostriches in South Africa, in order to determine the expected yield of by-products and saleable lean meat, fat and bone of rheas. 2. Differences (P < 0.05) between species were found in the proportional weight of the wings, feet, skin and liver. The wings, feet and head of rheas form a higher proportion of the carcase than in ostriches, whereas the skin of the former represents a lower percentage of body weight. 3. Lean meat production from rheas (64% on a carcase weight basis) is in the same order as for ostriches, broilers, turkeys and beef. PMID- 9347144 TI - Effect of clenbuterol on growth, carcase and skeletal muscle characteristics in broiler chickens. AB - 1. Male and female broiler chickens (144 in total) were given diets supplemented with clenbuterol (CB) at 0 (control) and at 1 mg/kg between 28 and 49 d of age to study the effect of CB on growth, carcase and skeletal muscle. 2. CB improved growth in males by increasing daily weight gain and final live weight and by lowering food conversion ratio. In females it changed the carcase composition by reducing abdominal fat pad and by increasing the proportion of protein. Consequently, carcase protein gain was increased in both sexes (11% and 16%, respectively). 3. Skeletal muscle weights were enhanced by between 6% and 22%. Muscle fibre diameters were increased in extensor hallucis longus (EHL) but not in gastrocnemius (GAS) muscle. This increase was more pronounced in females. EHL total muscle fibre number remained unchanged. The proportion of fast-twitch glycolytic fibres was increased at the expense of fast-twitch oxidative fibres in males only. Nuclear/cytoplasm and DNA/protein ratios tended to be decreased by CB. 4. From the elevated EHL muscle RNA/DNA, unchanged protein/RNA and translation activity it is suggested that CB stimulated protein synthesis at the pretranslational level. Reduced protein degradation is deduced from decreased neutral calcium-dependent proteolytic activity. 5. It is concluded that broiler chickens respond to long-term CB treatment as has been shown in various mammals. However, the sex-specific response in growth, carcase composition and skeletal muscle cellularity is more clearly apparent in broiler chickens. PMID- 9347147 TI - Food restriction and development of thermoregulation in Muscovy ducklings (Cairina moschata). AB - 1. The interaction between the effects of food restriction and cold stress on the development of body temperature, homeothermy index, metabolic rate and body weight were studied in Muscovy ducklings from hatching to 21 d of age. 2. The control group (ad libitum fed) and the food restricted group (fed to zero growth rate for 9 d) both became homeothermic when they were 2 d old with moderate (10 degrees C) cold stress. At severe cold stress (0 degrees C) the control group was homeothermic 5 days after hatching. However the food restricted group did not reach homeothermy at 0 degrees C and showed a large decrease (to hatching level) in homeothermy index at 10 degrees C and 0 degrees C after 9 d of food restriction. 3. Body temperature was lower in the food-restricted group during restriction and increased by 1 degree C after 24 h of ad libitum feeding. During food restriction, resting metabolic rate did not increase with age and was lower than the basal and existence metabolic rate predicted by Aschoff and Pohl (1970) and Kendeigh (1970) respectively. 4. The ratio of metabolisable energy (ME) intake to resting metabolic rate was 3 times lower in the food-restricted group than in the control group (0.09 and 0.27 respectively) on day 9. The availability of ME was more important than age for the development of thermoregulation in Muscovy ducklings. It is concluded that small improvements in the feeding regimen of young ducklings enhance the endurance and consequently reduce mortality from to environmental cold stress in a scavenger poultry system. PMID- 9347148 TI - Effect of soaking, germination, and enzyme treatment of whole barley on nutritional value and digestive tract parameters of broiler chickens. AB - 1. An experiment was carried out to determine the effect of soaking at 0 degrees C, soaking at room temperature, germination, or enzyme treatment of whole barley on feeding value and digestive tract parameters of 2- to 4-week old broiler chickens given diets with 700g/kg whole barley. 2. Soaking or germination decreased the soluble and total beta-glucan content (P < 0.05) and, except for soaking at 0 degrees C, the acid extract viscosity of the grain also decreased (P < 0.05). Germination and soaking in the presence of enzymes produced the lowest beta-glucan content and viscosity. 3. Except for soaking in cold water, the soaking, germination and enzyme treatments increased weight gain and decreased food:gain ratio (P < 0.05). Correspondingly, the digestibility of protein, fat, and ash, and the digestible energy content, increased (P < 0.05) after enzyme treatment or germination. 4. Chickens fed on enzyme-treated or germinated barley diets had intestinal contents with a greater proportion of dry matter and lower viscosity than chickens fed on untreated barley (P < 0.05). Consequently, the cages and chickens were cleaner (P < 0.05) and the weight of digestive organs as proportion of live weight was lower. 5. Particle size analysis of excreta revealed that whole barley was efficiently ground by the gizzards of 16-d-old chickens, and very few whole kernels were found. PMID- 9347149 TI - Performance, breast meat yield and abdominal fat deposition of male broiler chickens fed diets supplemented with DL-methionine or DL-methionine hydroxy analogue free acid. AB - 1. An experiment was conducted to compare the relative bioefficacy of DL methionine hydroxy analogue free acid (DL-MHA-FA) with DL-methionine in broiler chickens. Responses used for comparison were weight gain and food efficiency between 7 and 35 d of age, and breast meat deposition, food cost per kg of breast meat, and abdominal fat at 41 d of age. 2. A total of 2160 seven-day-old male broiler chicks were used. The feeding programme consisted of a starter diet from 7 to 21 d, and a finisher diet till the end of the experiment. The starter basal diet contained 6.1 g/kg total sulphur-containing amino acids (TSAA), and an estimated metabolisable energy (ME) content of 13.2 MJ/kg. The finisher diet contained 5.8g/kg TSAA and an estimated ME content of 13.6 MJ/kg. Four concentrations of DL-methionine and DL-MHA-FA were added at 0.5 g/kg increments on an equimolar basis. Therefore, there were 9 experimental treatments which were each applied to 6 replicates of 40 chicks. Weight gain and food efficiency were determined at 35 d of age. Breast yield and carcase fat were measured at 41 d. 3. Significant responses to graded amounts of both methionine sources were observed in weight gain, food efficiency, breast meat percentage, and food cost per kg of breast meat. The responses fitted exponential regression curves. Based on the regression coefficients, equimolar bioefficacy of DL-MHA-FA relative to DL methionine was 80% for daily gain, 83% for food efficiency, 51% for breast meat yield, and 66% for food cost per kg of breast meat. Differences between the 2 sources were significant (P < 0.05) for breast meat yield and food cost per kg of meat and (P < 0.10) for food efficiency. PMID- 9347150 TI - Effects of amino acid balance and energy:protein ratio on energy and nitrogen metabolism in male broiler chickens. AB - 1. An experiment was performed with growing broiler chickens (14 to 21 d old) to examine 3 diet characteristics which have been implicated in regulatory elevation of metabolic rate: an imbalanced amino acid mixture, high dietary energy concentration and low protein concentration. 2. Differences in energy expenditure could be explained almost entirely (93%) by differences in quantities, and therefore costs, of protein and fat accretion. There was no indication of regulatory diet-induced thermogenesis. Heat production was not significantly correlated with CP:TME ratio and was negatively correlated (P < 0.01) with dietary TME concentration. 3. Heat production was closely correlated (P < 0.001) with rate of protein accretion, which in turn was more strongly associated with intake of the first-limiting amino acid (lysine) than with total protein intake. Heat production on an imbalanced, lysine-limited, amino acid mixture was no greater than on a balanced amino acid source with the same lysine concentration. 4. There was no indication of a stimulation of heat production by excess amino acids. Heat production, adjusted for body weight by covariance analysis, was similar on paired diets which had identical lysine concentrations but a 1.5- or 2 fold difference in crude protein concentration. 5. There was a strong negative correlation (P < 0.001) between protein retention per g of lysine consumed and lysine: CP ratio, suggesting that, in this case, response to a limiting amino acid was improved by the presence of a super-abundance of other amino acids. PMID- 9347151 TI - Effect of temperature of wet extrusion on the nutritional value of full-fat soyabeans for broiler chickens. AB - 1. An experiment was conducted to determine the temperature for wet extrusion of full-fat soyabeans (FFS) needed to produce maximum chicken performance. 2. FFS were either unprocessed or extruded at 5 different temperatures (118 degrees, 120 degrees, 122 degrees, 126 degrees and 140 degrees C) in a wet extruder. Diets were prepared with the different FFS, and a diet prepared with soyabean meal (SBM) was included as a control. The 7 experimental diets were fed to individual groups of 40 chickens each, for a period of 35 d. Trypsin inhibitor activity (TIA), urease activity (UA), and protein solubility in potassium hydroxide (PS) were measured in all FFS and in the SBM. 3. Diets prepared with raw FFS and FFS extruded at 118 degrees and 120 degrees C resulted in significantly lower body weights and in pancreatic hypertrophy; maximum growth rate was obtained with FFS extruded at 122 degrees and 126 degrees C, while minimum pancreas weight was seen in chickens fed FFS extruded at 140 degrees C. 4. Although TIA, UA, and PS all decreased with increasing temperature of extrusion, TIA provided the best prediction of the feeding value of soyabeans for chickens. PMID- 9347152 TI - Studies on effects of nutritional factors on bone structure and osteoporosis in laying hens. AB - 1. A modern hybrid strain of laying hen (Hisex) was fed from point of lay to 68 weeks on a control diet and diets containing oystershell, fluoride, 1,25 dihydroxycholecalciferol, ascorbic acid, a lower concentration of phosphorus and a combination of a lower concentration of crude protein and higher concentration of vitamin K. Hens from a much older strain (Brown Leghorn J-line) were fed on the control diet. 2. Plasma variables were measured during lay. End-of-lay trabecular and medullary bone volumes in the proximal tarsometatarsus and free thoracic vertebra were measured by histomorphometry. 3. The majority of Hisex hens were considered to be osteoporotic by the end of lay. In contrast, none of the J-line were osteoporotic. 4. None of the nutritional treatments affected trabecular bone volumes. Medullary bone volumes were increased significantly by feeding oystershell or fluoride. 5. There was no phenotypic correlation between egg production and trabecular bone volume in the Hisex hens. 6. The experiment provided evidence that osteoporosis in laying hens, as assessed by trabecular bone volumes, is not caused by calcium deficiency and could not be prevented by any of the nutritional treatments studied. PMID- 9347153 TI - Tissue concentrations and pharmacokinetics of florfenicol in broiler chickens. AB - 1. Florfenicol (30 mg/kg body weight) was administered to broiler chickens via intravenous (i.v.), intramuscular (i.m.) and oral routes to study its plasma concentrations, kinetic behaviour, systemic bioavailability and tissue content. 2. Following a single i.v. injection, the kinetic disposition of florfenicol followed a 2-compartmental open model with an elimination half-life of 173 min, total body clearance of 26.9 ml/kg/min and a steady state volume of distribution of 5.11 l/kg. 3. The highest plasma concentrations of florfenicol were 3.82 and 3.20 micrograms/ml following single i.m. and oral administration, respectively. The systemic bioavailability was 96.6% and 55.3% after i.m. and oral administration. The plasma protein binding of florfenicol was 18.5%. 4. Following its administration, the highest tissue concentrations of the drug were found in the kidney bile, lung, muscle, intestine, heart, liver, spleen and plasma. Low concentrations of the drug were found in brain, bone marrow and fat. No florfenicol residues were detected in tissues and plasma after 72 h except in the bile from where it disappeared after 96 h. PMID- 9347154 TI - Physiological inhibition of growth hormone secretion by both insulin-like growth factors-I and -II in chickens. AB - 1. The role of both insulin-like growth factors (IGF)-I and -II in regulation of growth hormone (GH) secretion in chickens was examined. Seven-week-old male broiler chickens were injected intravenously (i.v.) with recombinant human IGF-I or IGF-II or specific anti-IGF-I or IGF-II immunoglobulins. Blood samples were taken before treatment and at 15 min intervals afterwards for 1 h. Controls received saline i.v. 2. Both IGF-I and IGF-II administration resulted in a rapid, significant decrease in plasma GH concentrations, but the concentrations of both triiodothyronine and thyroxine remained unchanged. 3. Immunisation against both IGF-I and IGF-II produced a significant elevation in plasma GH. 4. These data show that both IGFs can regulate GH concentrations in birds. Furthermore, the immunoneutralisation data suggest that these hormones have a physiological role in the regulation of GH secretion. PMID- 9347155 TI - Effect of ketone bodies on crop emptying in the chicken. AB - 1. The effect of ketone bodies on crop emptying was studied in chickens in 2 experiments. In the first, the effect of beta-hydroxybutyrate, acetoacetate or acetone on relative crop content was measured. The effects of dietary medium and long chain triacylglycerols upon serum beta-hydroxybutyrate were investigated in the second. 2. beta-Hydroxybutyrate, but not acetoacetate and acetone, delayed crop emptying in a dose dependent fashion. Serum beta-hydroxybutyrate concentration was high in chicks given medium chain triacylglycerol, when compared with long chain triacylglycerol. 3. The results suggest that delayed crop emptying induced by medium chain triacylglycerol could partly be explained by an enhanced concentration of serum beta-hydroxybutyrate, which is the result of the rapid oxidation of medium chain fatty acids. PMID- 9347156 TI - Effects of dietary cholestyramine on the utilisation of diets containing medium or long chain triacylglycerols by chicks. AB - 1. The effects of graded dietary concentrations of cholestyramine (CSTY, a bile acid binding polymer), which prevents micelle formation and bile acid reabsorption, on the lipid and energy metabolism of chicks given diets containing different dietary concentrations of medium chain triacylglycerol (MCT) and long chain triacylglycerol (LCT) were investigated. 2. MCT- or LCT-supplemented diets containing 100 or 200 g oil/kg diet and 0, 10 or 20 g CSTY kg were fed to 7 d old chicks for 10 d. As dietary CSTY concentration increased, a reduction in the metabolisable energy value was observed for both dietary lipid sources. Consequently, fat and energy retentions were also reduced as the dietary CSTY content increased. PMID- 9347157 TI - Ligation of caeca improves nitrogen utilisation and decreases urinary uric acid excretion in chickens fed on a low protein diet plus urea. AB - 1. The effect of the ligation of the caeca on nitrogen utilisation and nitrogen excretion was examined in conventional chickens fed a diet containing 50 g protein/kg plus urea. 2. Ligation of the caeca significantly improved nitrogen balance and utilisation by up to more than 2 times as much as those of controls (P < 0.05). 3. The treatment significantly decreased uric acid excretion by 77 mg nitrogen/day (P < 0.01) and also total nitrogen excretion (P < 0.05): the former decrease almost explained the latter. 4. No effect of the ligation of caeca on urea and ammonia excretion was observed. 5. It is concluded that nitrogen metabolism in chickens is affected by possible changes in caecal fermentation by preventing entry into the caeca of substances from urine and digesta. PMID- 9347158 TI - Moderate excess of dietary vitamin E does not exacerbate cholecalciferol deficiency in young broiler chicks. AB - 1. The combined effect of moderate excess dietary vitamin E and marginal amounts of dietary cholecalciferol on the performance and tibia bone ash of young male broiler chicks was evaluated. Vitamin E (alpha-tocopheryl acetate) and cholecalciferol were added to a commercial diet not already supplemented with these vitamins, at concentrations of 0 and 150 mg/kg, and 1.875, 5 and 25 micrograms/kg, respectively, and fed to chicks for 23 d. 2. Vitamin E concentration and its combinations with cholecalciferol did not significantly (P > 0.05) affect food intake, weight gain, food efficiency and bone ash. These variables were significantly (P < 0.001) lower in chicks fed on the diets supplemented with 1.875 micrograms cholecalciferol/kg compared with the values observed with the 2 other concentrations of this vitamin. There were no differences in the effects of 5 and 25 micrograms cholecalciferol/kg diet on the above variables. 3. It was concluded that vitamin E, at a concentration of 150 mg/kg diet, did not aggravate a mild cholecalciferol deficiency. PMID- 9347159 TI - A study on the growth curve of and maximum profit from layer-type cockerel chicks. AB - 1. 2900 commercial layer-type cockerel chicks were reared on the floor from 1-day old to 9 weeks of age. 2. The growth curve of the cockerel chicks was [formula see text] 3. The feeding costs (US$) of layer-type cockerel chicks were described by the equation Y = a + bx + cx2 = 0.0657 - 0.0091x + 0.0069x2. 4. When the layer type cockerel chicks' marketing price was US$0.82 per kg. (6.8 Renminbi per kg), the optimum marketing age for maximum profit margin was 5.9 weeks (41 to 42 d). PMID- 9347160 TI - Antibiotic prophylaxis for gastrointestinal endoscopy. PMID- 9347161 TI - Azathioprine therapy. PMID- 9347162 TI - Duodenal obstruction by a gallstone (Bouveret's syndrome) managed by endoscopic stone extraction: a case report and review. AB - Gastric outlet obstruction caused by a large gallstone in the duodenum or pylorus (Bouveret's syndrome) is a rare complication of gallstone disease. The presenting symptoms are often nonspecific and include nausea, vomiting, epigastric pain and a history of gallbladder disease. Although the diagnosis is established only at surgery in many cases, preoperative recognition by imaging techniques and endoscopy is desirable. Surgical treatment aims at removal of the ectopic gallstone, closure of the fistula and cholecystectomy. A case of Bouveret's syndrome is presented where endoscopic extraction of the duodenal gallstone was accomplished providing definitive treatment for this patient. PMID- 9347163 TI - Keeping an eye on Crohn's disease: orbital myositis as the presenting symptom. AB - Episodic periorbital swelling due to presumed orbital inflammation and myositis caused intermittent apparent proptosis and was the presenting symptom of ileocecal Crohn's disease (CD) in a teenage female with a family history of autoimmune disorders and CD. Orbital myositis, a very rare extraintestinal manifestation of inflammatory bowel disease (IBD), likely represents a process of impaired immunoregulation related to the underlying intestinal inflammation. This rare manifestation of IBD simulates the more commonly encountered thyroid orbitopathy (ophthalmopathy), but IBD should be considered if all thyroid tests are negative. It is important to recognize that orbital myositis may be an extraintestinal manifestation of Crohn's disease so that the diagnosis can be made and appropriate therapy commenced. PMID- 9347164 TI - Successful therapy of refractory erythema nodosum associated with Crohn's disease using potassium iodide. AB - Erythema nodosum is a common extraintestinal manifestation of Crohn's disease. While mild skin involvement often responds to conservative management, severe or refractory cases may require systemic corticosteroid or immunosuppressive therapy. This report describes successful treatment of severe, refractory erythema nodosum associated with Crohn's colitis using oral potassium iodide. While the mechanism of action of this agent is poorly understood, it appears to be an effective and nontoxic therapy for Crohn's-related erythema nodosum and warrants further evaluation in a placebo controlled trial. PMID- 9347165 TI - Use of azathioprine for nongranulomatous ulcerative jejunoileitis. AB - Nongranulomatous ulcerative jejunoileitis (NGUJI) is a rare, often fatal disorder that produces multiple nonmalignant small bowel ulcerations. A 55-year-old woman with presumed celiac disease presented with steroid-refractory diarrhea, weight loss and abdominal pain. A laparotomy was performed to exclude the possibility of a lymphomatous disorder, and multiple nonmalignant small bowel ulcerations were discovered. Despite a combination of treatment with total parenteral nutrition (TPN) and prednisone 30 mg/day she continued to deteriorate. The addition of azathioprine to her treatment regimen resulted in marked clinical and biochemical improvement. Her enteroscopy normalized, and she was able to discontinue TPN and reduce her steroid requirements. Although azathioprine has been used occasionally to treat refractory sprue, there have been no reports of its use in NGUJI. In this case, azathioprine played a key role in the management of NGUJI and should be considered a treatment option for patients with this disorder. PMID- 9347167 TI - Small bowel review: Part I. AB - Significant advances have been made in the study of the small bowel. Part I of this two-part review of the small bowel examines carbohydrates, including brush border membrane hydrolysis and sugar transport; amino acids, dipeptides, proteins and food allergy, with a focus on glutamine, peptides and macromolecules, and nucleosides, nucleotides and polyamines; salt and water absorption, and diarrhea, including antidiarrheal therapy and oral rehydration treatment; lipids (digestion and absorption, fatty acid binding proteins, intracellular metabolism, lipoproteins and bile acids); and metals (eg, iron) and vitamins. PMID- 9347166 TI - Pilot study of ofloxacin and interferon-alpha combination therapy for chronic hepatitis C without sustained response to initial interferon administration. AB - A controlled trial comparing combination therapy with ofloxacin (OFLX) and interferon (IFN) versus IFN monotherapy was conducted in patients with chronic hepatitis C who failed IFN therapy. Twenty patients were assigned randomly to two groups. Equal doses of recombinant IFN alpha-2b were administered to each group for 24 weeks. For the IFN plus OFLX group, OFLX was administered for 12 weeks at a daily dose of 600 mg. Levels of hepatitis C virus RNA declined significantly from the first month after the start of IFN treatment compared with those before administration in both groups. Serum alanine aminotransferase levels were significantly lower in the IFN plus OFLX group at two and six months after the start of treatment than levels in the IFN group. The fraction of subjects whose levels of serum ALT normalized was also higher in the IFN plus OFLX group. Larger clinical trials should be undertaken. PMID- 9347168 TI - Canadian Association of Gastroenterology Practice Guideline for informed consent- gastrointestinal endoscopy. PMID- 9347169 TI - Canadian Association of Gastroenterology Practice Guideline for clinical competence in diagnostic and therapeutic endoscopic retrograde cholangiopancreatography. PMID- 9347171 TI - Alberta Society of Gastroenterology consensus statement: Helicobacter pylori in peptic ulcer disease. PMID- 9347170 TI - Screening urinary bile acids for genetic defects in bile acid synthesis? PMID- 9347172 TI - Consensus Statement on Helicobacter pylori infection and its management. PMID- 9347173 TI - The Second Canadian Consensus Conference on the Management of Patients with Gastroesophageal Reflux Disease. AB - The Second Canadian Consensus Conference on the Management of Patients with Gastroesophageal Reflux Disease (GERD) was organized by the Canadian Association of Gastroenterology to address major advances in the understanding of the pathophysiology of GERD, to review the new methods of investigation and therapy introduced since the first conference in 1992 and to examine the issue of relevant health economics. The changes that have taken place over the past four years have been sufficiently dramatic to necessitate reassessment of the recommendations made following the first conference. The second conference dealt with the investigation and treatment of uncomplicated GERD and the complex issues of esophageal and extraesophageal complications such as chest pain, Barrett's esophagus, and reflux-related pulmonary and laryngeal disorders. The role of laparoscopic surgery was also discussed. A decision tree for investigation and treatment of patients with GERD was developed. The 38 participants represented a broad spectrum of experience, location of practice and special interests. The distribution of participants conformed to the recommendations of the Canadian Medical Association guidelines for consensus documents in that there should be input from all possible interested parties. A list of the state-of-the-art lectures presented during the conference, the small group sessions, the session chairpersons and participants are appended to this document. CONCLUSIONS. UNCOMPLICATED GERD: GERD with alarm symptoms must be investigated immediately. There was no consensus about when to investigate uncomplicated GERD, ie, whether to perform endoscopy immediately or after initial therapy fails. There was controversy regarding 'step up' (H2 receptor antagonist [H2RA] or prokinetic [PK] first therapy) versus 'step down' therapy (proton pump inhibitor [PPI] first therapy). The majority decision was for short term 'step up' therapy and investigation if symptoms do not improve or recur. Maintenance therapy should be carried out with the initial therapy that was effective. H2RAs and PKs may suffice for maintenance therapy in milder GERD; however, for severe esophagitis, PPIs should be used. SURGERY: Indications for laparoscopic surgery should be the same as for conventional antireflux operations. NONCARDIAC ANGINA-LIKE CHEST PAIN: After exclusion of nonesophageal causes, the majority decided that eight weeks of therapy with a PPI should be performed, while some suggested work-up before a therapeutic test. In the absence of response or recurrence, esophagogastroduodenoscopy (EGD) and, depending on the circumstances, 24 h ambulatory pH/motility may be indicated. BARRETT'S ESOPHAGUS: Only patients who, in case of future discovery of cancer or dysplasia, are able or willing to undergo therapy should have surveillance. In the absence of dysplasia EGD should be performed every two years, and in the presence of mild dysplasia every three to six months. All agreed that for severe dysplasia, esophagectomy or poor risk patients, esophageal mucosal ablation is indicated. ESTRAESOPHAGEAL COMPLICATONS (EECs): Asthma, chronic cough and posterior laryngitis were considered EECs. Although PPIs may decrease symptoms, improvement alone is not diagnostic of the presence of EEC. Ambulatory pH studies with two pH probes or ambulatory pH/motility may be useful in establishing causation. HEALTH ECONOMICS: There are limited data for an economic comparison among the different drugs or between medical and surgical therapy. PMID- 9347174 TI - Guidelines of the previous consensus conference and recent developments. AB - This article reviews the major changes that have occurred since the last Canadian consensus conference on gastroesophageal reflux disease (GERD), which was held four years ago. There were developments in the understanding of the pathophysiology of this disease and improvements in the methods of its investigation and management. Esophageal and extraesophageal complications have also been better defined. Since 1992 new knowledge on nitric oxide has been gained and several new inflammatory cytokines have been developed. Improved understanding of the mechanism of the hypersensitive esophagus helped to explain the presence of clinical symptoms with normal endoscopic findings. This also improved interpretation of the role of 24 h pH monitoring and 24 h pH with motility recording. There is better understanding of the role of H2 receptor antagonists and prokinetics and an increasing confidence in the long term safety of proton pump inhibitors (PPIs). The most important changes occurred in surgery with the introduction of laparoscopic fundoplication. Patients with Barrett's esophagus who are too ill for esophagectomy may now be enrolled in surveillance because it is possible to deal with dysplasia and small cancers with photodynamic therapy. As an introductory lecture to the consensus conference, this article also deals with the subject of health economics and discusses the necessity and the dangers involved in devising treatment guidelines. It indicates that guidelines change with advancing knowledge, and emphasizes that physicians' primary duty is toward their patients. Therefore, guidelines devised here should be adjusted to the individual needs of patients. Organizational aspects of the present conference are also described. PMID- 9347175 TI - Esophagitis as the outcome of progressive failures of the defensive repertoire. AB - Pre-epithelial defences include the coordinated actions of the lower esophageal sphincter and the esophageal muscles, which minimize reflux of gastric contents and promote clearance of refluxed material. The esophageal epithelium also possesses innate resistance to luminal damaging agents and may be protected luminally by a mucus or 'mucus bicarbonate' barrier and possibly a layer of hydrophobic surfactants. These components are derived from submucosal glands located in the submucosal connective tissue and from salivary secretions that may bind to the esophageal surface. Epithelial defences include the glycocalyx, permeability properties of the epithelial cell plasma membrane, junctional barriers to proton permeation through the paracellular pathway and ion transport processes for regulation of intracellular pH. Subepithelial defences involve mainly regulation of blood supply via responses of nerves, mast cells and blood vessels to influxing protons. Although the epithelium can withstand prolonged exposure to physiologically relevant concentrations of acid, the presence of pepsin or bile salts may overcome the permeability barrier, which probably resides in the superficial layers of epithelial cells. Focal destruction of these cells allows access of luminal acid and other aggressive agents to the vulnerable basolateral cell membranes and to the submucosa. The result is lesion production, although an efflux of alkaline plasma may protect the underlying submucosa and allow healing. Salivary-derived epidermal growth factor (EGF) is present in the luminal fluid, and lesion development may also provide access of EGF to receptors within the epithelium and in the underlying vasculature. Accelerated cell proliferation would then contribute to healing. Inflammation and healing should also be viewed as defensive responses, as can the development of Barrett's esophagus, in which the stratified squamous epithelium is replaced by a potentially acid-resistant columnar epithelium. Chronic inflammation and esophagitis only result when this multilayered set of defences is overcome. The challenge for research is to identify those components of the defensive repertoire that are defective in individuals who suffer from chronic esophagitis. PMID- 9347176 TI - Assessment of clinical severity and investigation of uncomplicated gastroesophageal reflux disease and noncardiac angina-like chest pain. AB - Heartburn, suggesting gastroesophageal reflux, is common. Epidemiological studies have shown that 36% to 44% of adults experience heartburn at least once a month, 14% weekly and 7% once a day. Heartburn and regurgitations are the typical symptoms of gastroesophageal reflux disease (GERD). When present as predominant symptoms, they are quite specific but not very sensitive. Clinical severity of GERD does not predict the severity of the underlying condition. The diagnostic approach to patients with GERD depends on the clinical presentation and the question to be answered -Is abnormal reflux present? Is there mucosal injury? Are symptoms due to reflux? Several techniques such as barium swallow, endoscopy, ambulatory pH monitoring, esophageal manometry and 24 h pH/motility can be used to answer those questions. Barium swallow is not much help in diagnosing reflux esophagitis because reflux can occur in more than 25% of asymptomatic patients. It is most useful in demonstrating structural abnormalities such as strictures and hiatal hernia. The importance of hiatal hernia in the pathogenesis of GERD has been controversial. Recent studies suggest that GERD patients with hiatal hernia present with greater extent of reflux and more severe esophagitis. Endoscopy is the best diagnostic study for mucosal evaluation. Ambulatory 24 h pH monitoring is indicated for patients with atypical symptoms of reflux such as chest pain or pulmonary symptoms, or those who do no respond to therapy. The evaluation of duodenogastroesophageal reflux or alkaline reflux can be measured, but the clinical importance of this test remains controversial. Esophageal manometry allows measurement of the lower esophageal sphincter pressure (LES) and the evaluation of esophageal peristalsis. There is a lack of correlation between LES and reflux esophagitis. The role of peristaltic dysfunction in GERD is unclear, but the high percentage of abnormal contractions suggests a more severe form of GERD. Esophageal motility study can document the presence of effective esophageal peristalsis in patients before antireflux surgery. Twenty-four hour pH/motility is not yet available widely. It is useful in patients who have several daily attacks. There is a correlation with acid reflux in approximately 40% of events. Investigation of noncardiac angina-like chest pain is best achieved by standard esophageal manometry combined with provocative testing. Most laboratories performing these studies use acid perfusion and pharmacostimulation with either bethanechol or edrophonium to reproduce the patient's chest pain during esophageal manometry. Esophageal balloon distension is considered to give the highest yield as a provocative test in patients with angina-like chest pain. It is believed that abnormal esophageal nociception is not simply related to underlying motor dysfunction but also to the presence of a visceral sensory abnormality. PMID- 9347177 TI - Pathophysiology and investigation of Barrett's esophagus. AB - Barrett's esophagus is an acquired condition with columnar metaplasia of the distal esophagus. Gastroesophageal reflux is the main pathophysiological factor, although genetic predisposition may play a role. The significance of Barrett's esophagus is that it is the only recognized risk factor for adenocarcinoma of the esophagus, which is one of the most rapidly rising types of cancer in North America. Cancer develops in Barrett's esophagus through a series of steps including mucosal dysplasia. High grade dysplasia is clearly a premalignant lesion and has been the focus of endoscopic surveillance strategies. Because dysplasia and adenocarcinoma develop predominantly in patients with specialized intestinal columnar epithelium, they make up the group that would be considered for surveillance programs. Endoscopic surveillance for dysplasia is only indicated for patients in whom esophagectomy would be considered if high grade dysplasia or carcinoma was found, at least until other endoscopic ablative techniques are proven to be beneficial. It has been recommended that endoscopy be performed every other year and be increased to yearly if low grade dysplasia is found. High grade dysplasia should be confirmed by another expert pathologist, and the patient should then be considered for esophagectomy. Flow cytometry and genetic markers may improve the ability to select patients for surveillance programs in the near future. PMID- 9347178 TI - Extraesophageal complications of gastroesophageal reflux disease. AB - With the widespread availability of ambulatory esophageal pH monitoring, there has been recently renewed interest in the so-called 'extraesophageal' complications of gastroesophageal reflux disease (GERD). There are two proposed mechanisms by which reflux can cause extraesophageal symptoms or disease: refluxed acid may reach the oropharynx and/or respiratory tract and cause direct irritation; or acid contact with the esophageal mucosa may trigger neural reflexes, which, in turn, produce symptoms. Evidence is most compelling for an association between GERD and unexplained dental erosions, posterior laryngitis, chronic unexplained cough and intrinsic asthma. The clinician should be aware of these associations, and patients with these conditions should be questioned carefully about associated GERD symptoms. When GERD and any of these conditions coexist, intensive medical antireflux therapy is indicated. Twenty-four hour pH monitoring may be required in selected patients to document the relationship between reflux and the extraesophageal complication or to ensure that the medical therapy provided has eliminated acid reflux. PMID- 9347179 TI - Efficacy of H2 receptor antagonists in the treatment of gastroesophageal reflux disease and its symptoms. AB - Gastroesophageal reflux disease (GERD) is caused by prolonged esophageal mucosal exposure to acid gastric refluxate due to failure of the normal antireflux mechanisms of the lower esophageal sphincter. Gastroesophageal reflux can be controlled by suppression of acid secretion or by improvement of gastric emptying and esophageal clearance. H2 receptor antagonists are the most commonly used antisecretory drugs, and in the past 20 years have constituted the cornerstone of therapy for the treatment of reflux disease. They have been shown to be effective in the symptomatic treatment of intermittent or mild nonerosive GERD (greater than 70%). When used at the usual recommended dose, all four H2 receptor antagonists (cimetidine, ranitidine, famotidine and nizatidine) are equally effective and are found to be generally very safe; interactions with other drugs metabolized through the cytochrome p450 notwithstanding. However, their efficacy is limited in more severe forms of GERD such as erosive esophagitis (symptomatic improvement 40% to 60%, endoscopic healing 40% to 50%) in which the superior efficacy and more rapid symptomatic relief provided by proton pump inhibitors is clearly demonstrated. The availability of H2 receptor antagonists for over-the counter use will increase their use for mild and intermittent disease but is unlikely to alter the need for more potent acid suppression in aggressive reflux disease. PMID- 9347180 TI - Prokinetic therapy in gastroesophageal reflux disease. AB - There is a growing body of pathophysiological evidence that gastroesophageal reflux disease (GERD) is caused by disordered motility and not acid hypersecretion. The key factor in the pathogenesis of GERD is disordered function of the lower esophageal sphincter. Other factors include delayed gastric emptying and decreased peristalsis in the body of the esophagus. The principal symptoms of GERD are heartburn and regurgitation. Studies have demonstrated that up to 50% of patients may have other symptoms of dysmotility including epigastric discomfort or fullness, nausea and early satiety. The use of a prokinetic agent in such patients seems logical. Given its proven superior efficacy over domperidone and metaclopramide in treating GERD, cisapride has become the prokinetic drug of choice for the acute management and maintenance therapy of GERD. In the acute management of GERD, cisapride is superior to placebo and has the same efficacy as H2 receptor antagonists (H2RAs) in several clinical trials. It is also effective in maintenance therapy for GERD. These studies are reviewed. Cisapride (10 mg qid or 20 mg bid) is effective in the acute treatment of mild to moderate GERD, particularly in patients with heartburn associated with other symptoms of dysmotility, and particularly in patients with heartburn associated with gastroparesis. Combination therapy with an H2RA may be considered if symptoms (particularly dysmotility symptoms) persist with H2RA alone. In severe GERD that is not responsive to conventional doses of a proton pump inhibitor, cotherapy with cisapride or increasing the dose of the proton pump inhibitor are the two therapeutic options to consider. Cisapride 20 mg at bedtime is effective maintenance therapy for patients with mild to moderate GERD. PMID- 9347181 TI - Proton pump inhibitors in acute healing and maintenance of erosive or worse esophagitis: a systematic overview. AB - The aim of this paper is to present a systematic overview of the efficacy of the proton pump inhibitors (PPI), omeprazole and the newer lansoprazole, in the healing and maintenance of erosive or worse (grade II to IV) esophagitis. At the time of the 1996 gastroesophageal reflux disease (GERD) consensus meeting, a third PPI, pantoprazole, was not yet available in Canada and was, therefore, not discussed. The present review is therefore restricted to patients with more severe disease than that of the average patient who presents to the family physician with symptoms of mild GERD. In these patients with endoscopic evidence of damaged esophageal mucosa, both proton pump inhibitors were highly effective and safe in acute healing of erosive esophagitis and were significantly better than H2 receptor antagonists, healing faster and much more completely, with shorter durations of treatment. Suggested initial doses were omeprazole 20 mg once daily and lansoprazole 30 mg once daily, which were consistent with the manufacturer's recommendations. Once patients with this degree of esophagitis have their mucosal lesions healed, they almost inevitably have recurrence of esophagitis (80% at one year) unless some form of maintenance therapy is continued. Unfortunately, for these patients with healed erosive or ulcerative esophagitis, H2 receptor antagonists appear to be no better than placebo, and a PPI is the only class of drug able to minimize relapse significantly. PMID- 9347182 TI - Role of surgery in the management of gastroesophageal reflux disease. AB - Surgical management of gastroesophageal reflux disease (GERD) can provide good long term control of symptoms. The introduction and increasing use of laparoscopic antireflux procedures may provide an early surgical alternative to long term medical control of GERD. Indications for surgery, preoperative investigations, surgical options, and results and complications are discussed. PMID- 9347183 TI - Value of a therapeutic trial to diagnose gastroesophageal reflux disease: step up versus step down therapy. AB - The 1992 Canadian Association of Gastroenterology consensus conference on gastroesophageal reflux disease (GERD) recommended a therapeutic trial of H2 receptor antagonists as a cost effective approach to the management of patients with typical symptoms of GERD. Omeprazole, with its increased potency and efficacy of acid suppression, appears to be the ideal diagnostic test therapy for GERD. However, there is little evidence in the literature to support this approach. Omeprazole has the potential to mask other disorders such as peptic ulcer disease, thereby delaying appropriate testing for and eradication of Helicobacter pylori. Therefore, omeprazole therapeutic trials should be used with caution in the diagnosis of GERD. The conventional step up therapy is the least costly and reasonably effective approach to treat the majority of patients with mild to moderate symptoms of GERD. For patients who fail therapy or have complications of GERD, the recently proposed step down therapy is probably more effective and appropriate. As physicians become more comfortable with the long term use of proton pump inhibitors, step down therapy may well replace the more conservative step up approach to therapy. PMID- 9347184 TI - Value of gastroscopy without a prior consultation. AB - Upper endoscopy is the most accurate and cost effective tool available to physicians when confronted with a patient with dyspepsia. Access to this procedure in Canada is generally limited by the requirement to see a consultant physician before the procedure can be performed. This review, in a question and answer style, builds a case for the introduction of open access endoscopy in Canada. Endoscopy is an effective and valid diagnostic test. Consultants are no better at deciding who requires endoscopy than general practitioners. Clinical patterns of dyspepsia are not a valid guideline for appropriate use of endoscopy. There is value in a normal endoscopy in terms of changes to medical treatment, reassurance, lower consultation and medical clinic attendance rates--and radiation exposure is avoided. After endoscopy, normal patients tend to make 41% fewer physician visits, prescriptions decrease by 71%, and 66% feel better and are more productive at work. Open access endoscopy is cost effective, even when American costs are used. Open access endoscopy should not overrun endoscopy suites if we use the British experience as a guide. The endoscopist would only require a limited history and physical examination to determine safety of the procedure. Follow-up and management is the responsibility of the referring physician, and this must be communicated to the patient and referring physician. The Canadian Association of Gastroenterology needs to draw up guidelines along with the Canadian Medical Practitioners Association for the adoption of this practice. PMID- 9347185 TI - Controversial issues in gastroesophageal reflux disease. AB - The management of gastroesophageal reflux disease (GERD) has advanced dramatically in the past 20 years because of increased understanding of its pathophysiology and improvements in available treatments. A number of important new ideas arousing controversy include the place of over-the-counter H2 receptor antagonists; the role of endoscopy in diagnosis of reflux symptoms; initial treatment options with regard to stepping up therapy to the most effective as needed or stepping down therapy from a proton pump inhibitor given initially because of its superior efficacy in all grades of reflux disease; the optimal choice of long term management of patients with GERD or GERD complicated by Barrett's metaplasia; the risk of Barrett's metaplasia; and the role of surveillance of this population. This paper reviews these topics and addresses the controversial issues. PMID- 9347186 TI - Management of complicated gastroesophageal reflux disease: atypical chest pain. AB - Although most patients with gastroesophageal reflux disease (GERD) present with the classic symptoms of heartburn and acid regurgitation, many complain of atypical chest pain suggestive of cardiac disease. Once cardiac ischemia has been excluded, it is important to consider GERD because this may be established as the cause of pain in 10% to 50% of such patients. If GERD is suspected or documented, vigorous antireflux treatment, preferably with proton pump inhibitory therapy, is indicated. PMID- 9347187 TI - The treatment of peptic esophageal strictures. AB - Symptomatic stricturing of the esophagus complicates the course of about 10% to 15% of patients with gastroesophageal reflux disease, particularly if they are elderly or if there is an associated Barrett's esophagus. Treatment goals include relief of symptoms of reflux disease and dysphagia, and prevention of their recurrence. The main therapeutic option to date has been endoscopic dilation. Although more than 30% of patients require serial long term dilations, this proportion can be minimized by the concomitant use of long term, high dose proton pump inhibition. Indications for surgery include failure of medical management. It is too early to assess the impact of laparoscopic technology on the treatment of peptic strictures. At this time, well designed prospective comparative trials are needed to quantify better the cost effectiveness of available treatment strategies in the management of patients with esophageal peptic strictures. PMID- 9347188 TI - Treatment of Barrett's esophagus. AB - Barrett's esophagus represents the most serious consequence of chronic gastroesophageal reflux disease (GERD), primarily because of its association with an increased incidence of esophageal adenocarcinoma. Specific therapy for Barrett's esophagus should lead to the complete regression of the metaplastic epithelium with adequate squamous reepithelialization. Ideally, this regression should be permanent and be associated with a reduction in the incidence of adenocarcinoma. Several reports in the literature have assessed the effects of H2 blocker treatment of Barrett's epithelium, but none has clearly documented a significant and consistent regression of the metaplastic epithelium. Proton pump inhibitors have been shown to be superior to H2 blockers in the treatment of patients with severe esophagitis. Despite initial enthusiasm, it does not appear that a significant regression of Barrett's epithelium can be achieved, even with high doses of proton pump inhibitors given for a prolonged period of time. Various groups have assessed the effects of antireflux surgery on the regression of columnar epithelium and dysplasia and its potential protective effect on the subsequent development of carcinoma. Overall, it appears from these reports that antireflux surgery, despite adequate symptomatic results, does not significantly and consistently lead to a reduction in length or disappearance of the Barrett's mucosa, and does not prevent the development of dysplasia and its progression to carcinoma. More recently, numerous authors have documented the regression of Barrett's mucosa by using various endoscopic thermal modalities. Technological advances including laser and photodynamic therapy have allowed for endoscopic mucosal ablation. Long term results are more encouraging when this mucosal ablation is associated with aggressive antireflux therapy (medical or surgical). Further studies are required before these exciting new therapies can be recommended. Currently, none of these approaches can obviate the need for continued endoscopic surveillance. PMID- 9347189 TI - Treatment of extraesophageal manifestations of gastroesophageal reflux disease. AB - Extraesophageal manifestations of gastroesophageal reflux disease (GERD) include chronic cough, asthma and 'acid' laryngitis. The response to medical and/or surgical therapy of these conditions is highly variable and often delayed. Of patients with GERD-related symptoms, those with extraesophageal manifestations are some of the most difficult to treat. Histamine antagonists, proton pump inhibitors and antireflux surgery have all been used to treat GERD-related asthma with variable results. Asthma patients who do not respond to high-dose acid suppression may be refractory to all forms of therapy. GERD is the third most common cause of chronic cough, and therapeutic results with acid suppression and antireflux surgery are variable. Posterior laryngitis presents as chronic hoarseness and has been shown to resolve clinically and histologically with acid suppression therapy or antireflux surgery. Results are variable, and controlled trials are lacking. PMID- 9347190 TI - Health economics: a clinician's perspective. AB - Health care economics is primarily concerned with the study of scarcity and choice. This has become increasingly important as health care spending is reduced. Reductions in spending are driven by federal and provincial government deficit reduction. Health economics analysis can be applied from a number of perspectives. Many physicians perceive health economics as a cost cutting exercise not primarily concerned with the quality of patient care. Health economics analysis may lead to ethical dilemmas in the physician/patient relationship. The physician's obligation to maintain an ethical relationship with patients may conflict with his or her role as rationer of health resources. It is difficult to balance conflicting roles, but physicians are obliged to participate in the economic debate to ensure optimal patient care. PMID- 9347191 TI - Health economics of gastroesophageal reflux disease. AB - The present study provides an overview of the current state of health economics studies of gastroesophageal reflux disease (GERD). It indicates the strengths and weaknesses of individual studies, and the state of health economics analysis in general as they apply to GERD. Specifically, this study adopts a pharmacoeconomic perspective, which is a subsection of health economics analytical methods, to provide a comparative analysis of alternative courses of action based on cost and consequence. The pharmacoeconomic outlook is most effective when it considers a comprehensive societal perspective, with special consideration given to other relevant viewpoints, such as the payer, the primary provider and, most important, the patient. Pharmacoeconomics provides several specific analytical techniques for GERD-related health economics analysis. The Canadian Association of Gastroenterology consensus conference on GERD in 1996 thought that a cost effective analysis was the most appropriate technique to assess the pharmacoeconomics of GERD. Six previous studies on GERD health economics have been performed comparing omeprazole with H2 receptor antagonists. These studies vary in cost data collected and in analytical techniques. In general, the existing outcome measurements of these previous health economics studies are not ideal. Namely, they combine various GERD grades, use randomized controls, are endoscopically based, assess pharmaceutical therapy only and are short term. More appropriate health economic trials in GERD, which focus on GERD management strategies and therapeutic treatment of GERD, need to be designed and conducted. These economic assessments, however, should not replace detailed thinking, careful observation, good judgement and common sense. PMID- 9347192 TI - Muscle oxygen saturation during surgery in the lithotomy position. AB - Surgery in the lithotomy position can provoke ischaemic lesions in the lower leg. We assessed lower leg oxygen saturation using near-infrared spectroscopy (NIRS) in 42 patients undergoing urinary tract surgery. Lower leg perfusion pressure was calculated as the difference of mean arterial pressure to pressure in an air bag supporting the lower leg and the hydrostatic pressure difference from the level of the lower leg to the heart. During elevation of the lower leg for 25 (3-65) min (median and range), mean arterial pressure decreased from 100 (73-125) to 77 (53-112) mmHg and the lower leg perfusion pressure dropped from 103 (80-122) to 21 (-6-65) mmHg, corresponding to a reduction in oxygen saturation of the medial gastrocnemius muscle from 68% (40-100%) to 58% (20-96%) (P < 0.01). The results demonstrate significant desaturation of the calf muscles during surgery in the lithomy position. PMID- 9347193 TI - Does long-term smoking affect aortic stiffness more in women than in men? AB - Smoking is a well-known risk factor for cardiovascular disease, although understanding of the pathophysiological mechanism is incomplete. The effect of heavy smoking, for more than 20 years and of 20 cigarettes per day, on aortic stiffness was studied in women (n = 23, age range 43-61 years) and men (n = 21, age range 43-61 years) who smoked but were otherwise healthy and compared with a healthy reference population that did not smoke. Aortic stiffness (beta) was calculated from the diameter and the pulsatile diameter change determined non invasively using an ultrasonic echo-tracking system and blood pressure obtained by the auscultatory method. The results showed that aortic diameter did not differ in smoking males (P = 0.974) or in smoking females (P = 0.361). Aortic stiffness was increased in the female (P = 0.041) but not male smokers (P = 0.222). Systolic, mean and diastolic blood pressure in the men and women who smoked did not differ from those in the healthy reference population. In conclusion, this investigation shows increased aortic stiffness in female but not in male smokers. It indicates that the aorta of women might be more vulnerable to smoking with regard to stiffening and degeneration than the aorta of men. PMID- 9347194 TI - Release of atrial natriuretic peptide during pulmonary artery clamping in man. AB - The vasodilating hormone atrial natriuretic peptide (ANP) is secreted from the heart in response to atrial wall stretch, but knowledge of the time course of ANP secretion after acute releasing stimuli is limited. The time from stimulus to release of immunoreactive atrial natriuretic peptide (irANP) was investigated by unilaterally clamping the pulmonary artery in 12 patients undergoing pulmonary surgery. The second messenger to ANP-induced vascular relaxation, plasma cyclic guanosine monophosphate (p-cGMP), was measured as an indirect marker of the vascular effects of ANP. Immediately after applied clamping, p-irANP (baseline level 15.4 +/- 2.9 pmol l-1) started to increase, reaching a significant, although moderate, increase (11 +/- 4%, P < 0.05) after 2 min. This elevation of p-irANP remained during the entire clamping period (+13 +/- 6%, P < 0.05 vs. baseline). Within 1 min of declamping, p-irANP returned to the baseline level. No conclusive alterations in p-cGMP were observed. The prompt ANP response to the applied stimulus, and the return to baseline after declamping, may indicate the presence of a short time-acting releasing mechanism of ANP. PMID- 9347195 TI - Effects of strength and endurance training on muscle fibre characteristics in elderly women. AB - The effects of 18 weeks' intensive strength and endurance training on fibre characteristics of the vastus lateralis muscle were studied in 76- to 78-year-old women. Type I and type IIa fibres constituted over 90% of the cell population and were almost equally represented. No changes were observed in the proportions of the different fibre types. When comparing the baseline and the 18-week measurements within the groups, the strength group showed a mean increase of 34% (P = 0.028) in mean type I fibre area. The frequency histograms showed an increased proportion of larger type I fibres after strength training and a decreased proportion of smaller type IIa fibres after endurance training. In the control subjects, the proportion of smaller type I and type IIa fibres increased during the experimental period. The results indicate that intensive strength training induces type I fibre hypertrophy, whereas the effects of endurance training are less evident. The considerable variation found in the change in muscle fibre cross-sectional areas is also noteworthy. PMID- 9347196 TI - Potassium and ventilation during positive and negative work in patients with chronic obstructive pulmonary disease. AB - In patients with chronic obstructive pulmonary disease (COPD), reduced ventilatory reserves limit exercise tolerance. In these patients, the ventilatory requirements of eccentric exercise (negative work, Wneg) are lower than those of concentric exercise (positive work, Wpos) at similar workloads. In this study, we investigated the relationship between plasma potassium levels and ventilation during Wpos and Wneg in these patients. Twelve patients with stable COPD [mean (SD) FEV1 46% (16) of predicted] performed Wpos and Wneg on a cycle ergometer (6 min of exercise; interval > or = 1 h) in a randomized order at a constant workload of 50% of the individual maximum (positive) work capacity. Minute ventilation (VE) and arterial plasma potassium concentration ([K+]a) were measured at rest, and at 1-min intervals during exercise and during 3 min of recovery. VE increased less during Wneg than during Wpos [6 (range 3-26) vs. 18 (range 8-28) 1 min-1; P < 0.01]. VE during Wneg was reduced in proportion to VCO2. The increase in [K+]a during Wpos and Wneg [0.45 (range 0.26-0.75) and 0.34 (range 0.1-0.97) mM] did not differ significantly. VE was closely correlated with VCO2 during both types of exercise. VE was also closely correlated with [K+]a, but the slope of the relationship between [K+]a and VE was steeper during Wpos than during Wneg [39.1 (range 15.2-88.6) vs. 18.3 (range 7.2-37.3) 1 min-1 mM-1; P = 0.012]. In contrast, the slope of the relationship between VCO2 and VE was similar during both types of exercise [27.8 (range 19.2-37.1) vs. 32.1 (range 19.8-48.4)]. Thus, for a given increase in [K+]a, the increase in VE was significantly less during Wneg. In patients with COPD, potassium did not explain the difference in exercise ventilation between Wneg and Wpos, and may not play a significant role in the control of breathing during low-intensity exercise. PMID- 9347197 TI - Mesenteric artery response to head-up tilt-induced central hypovolaemia and hypotension. AB - Superior mesenteric artery (SMA) blood flow and impedance were evaluated by duplex ultrasound during head-up tilt (HUT)-induced central hypovolaemia and hypotension in eight healthy volunteers. HUT induced a reduction in cardiac stroke volume from 88.8 +/- 6.3 to 64.7 +/- 6.3 ml (mean +/- SEM; P < 0.01) and an increase in thoracic electric impedance from 38.6 +/- 2.1 to 42.6 +/- 2.1 omega (P < 0.01) reflecting a reduced central blood volume. Maintained tilt provoked a 30% reduction in mean arterial pressure (from 87.1 +/- 3.3 to 63.4 +/- 3.6 mmHg: P < 0.01) and the appearance of presyncopal symptoms. During both the normotensive and the hypotensive phase of HUT, the SMA diameter (5.7 +/- 0.03 mm) and blood flow (514 +/- 75 ml min-1) did not change significantly, although the end-diastolic velocity increased from 9.7 +/- 4.8 to 39.7 +/- 4.0 cm s-1 (P < 0.01). The increase in diastolic velocity, despite a maintained or reduced arterial pressure, supports a reduction in the SMA impedance as it was reproduced during a meal test when a moderate reduction in mean arterial pressure (87 +/- 4 to 80 +/- 4 mm Hg; P = 0.04) was accompanied by a ninefold increase in the end diastolic velocity (P < 0.01). The results indicate a reduction in the mesenteric vascular impedance to the extent that superior mesenteric artery blood flow is maintained during HUT-induced central hypovolaemia and hypotension. PMID- 9347198 TI - Ventilatory response to CO2 in patients with snoring, obstructive hypopnoea and obstructive apnoea. AB - Obstructive sleep apnoea (OSA) is caused by an obstruction of the upper airway. Sufficient sensitivity to CO2 in the respiratory centre is known to be a critical factor for adequate tone in the upper airway muscles. The hypothesis of this study is, therefore, that the ventilatory response to CO2 is reduced in patients with OSA. Twenty-six patients who suffered from snoring, 19 snoring patients with obstructive hypopnoea (OH) and 33 snoring patients with obstructive apnoea (OA), were studied. The control group consisted of 25 subjects from a random sample with no history of snoring or daytime sleepiness. Tests of the hyperoxic and hypoxic ventilatory response to CO2 were performed, as well as static and dynamic spirometry. Subjects in the OA group displayed a higher hyperoxic (VE/FetCO2hy = 12.6 l min-1/%) and hypoxic (VE/FetCO2ho = 15.7 l min-1/%) ventilatory response to CO2 than patients with obstructive hypopnoea (VE/FetCO2hy = 8.6 l min-1/%; VE/FetCO2ho = 15.2 l min-1/%), snorers (VE/FetCO2hy = 8.4 l min-1/%; VE/FetCO2ho = 12.7 l min-1/%) and non-snorers (VE/FetCO2hy = 7.6 l min-1/%; VE/FetCOho = 9.6 l min-1/%). Multiple regression analysis reveals that neck circumference, apnoea index, oxygen desaturation index, PCO2 and sex (male gender) are correlated with VE/FetCO2hy (R2 = 0.43). Multiple regression analysis also reveals that ERV (expiratory reserve volume) and sex (male gender) are correlated with VE/FetCO2ho (R2 = 0.21). Arguing against the hypothesis, patients with OSA displayed an increased hyperoxic and hypoxic ventilatory response to CO2. Nocturnal apnoea frequency and the obesity factor in OSA may have contributed to these results. PMID- 9347199 TI - Effects of adrenergic blockade on hepatic glucose production during ethanol administration. AB - Acute ethanol administration stimulates sympathetic nervous system activity. The present study was designed to determine whether this sympathetic activation affects glycogenolysis and total hepatic glucose production (HGP) during ethanol induced inhibition of gluconeogenesis. Nineteen volunteers participated in four protocols. Two protocols aimed to study--using combined infusion of [6,6 2H2]glucose and [U-13C]glucose, VCO2 and 13CO2 measurements--the effects of ethanol infusion alone (n = 10) or with propranolol (n = 6) or phentolamine infusion (n = 4) on HGP, glucose disposal (Rd), glucose oxidation [13C]Glcox and non-oxidative glucose disposal (NOGD = Rd - [13C]Glcox). The fourth protocol assessed the effects of saline infusion alone on HGP. Using ethanol, HGP decreased by 23%, Rd by 20% and glycaemia by 9% (all P < 0.001); heart rate increased by 10%, whereas blood pressure remained unchanged. The effects were not observed with saline, except a slight (10%) decrease in HGP (P < 0.01 vs. ethanol). Ethanol did not affect [13C]Glcox but decreased NOGD by 73% (P < 0.001). Propranolol or phentolamine did not alter any of the effects of ethanol on glucose metabolism, but decreased mean arterial pressure. Propranolol prevented the ethanol-induced increase in heart rate. In conclusion, ethanol decreased blood glucose by decreasing HGP, presumably by inhibiting gluconeogenesis. Sympathetic activation prevented the decrease in blood pressure produced by ethanol but did not stimulate glycogenolysis. PMID- 9347200 TI - The twitch interpolation technique for the estimation of true quadriceps muscle strength. AB - The aim of this study was to examine the reliability of the twitch interpolation technique when used to estimate the true isometric knee extensor muscle strength. This included an examination of whether submaximal activation causes any bias in the estimation of the true muscle strength and an examination of the precision of the method. Twenty healthy subjects completed three contraction series, in which the subjects were told to perform as if their voluntary strength was 60%, 80% or 100% of that determined by a maximal voluntary contraction (MVC). Electrical muscle stimulations were given at each of five different contraction levels in each series. At torque levels above 25% of MVC the relationship between torque and twitch size could be approximated to be linear. The true muscle strength (TMS) could therefore be estimated using linear regression of the twitch-torque relationship to the torque point of no twitch in each of the three series, termed TMS60, TMS80 and TMS100. The TMS80 was slightly lower (P < 0.01), median 94% (IQ range 87-101%) of the TMS100. The TMS60 was median 99% (IQ range 83-125%) (NS) of TMS100, but a few severe outliers were observed. In conclusion, we found the reliability of the method acceptable for many research purposes, if series with estimated central activation of below 40-50% were excluded. The only moderate precision and the slightly lower estimations in subjects applying submaximal does, however, limit its usefulness. PMID- 9347201 TI - The effect of flumazenil on subclinical psychometric or neurophysiological alterations in cirrhotic patients: a double-blind placebo-controlled study. AB - It is not yet clear if benzodiazepine receptor ligands, implicated in the pathophysiology of hepatic coma, also have a role in subclinical cognitive or neurophysiological alterations in cirrhotic patients. Therefore, we carried out a double-blind, placebo-controlled study to evaluate the effectiveness of flumazenil, a benzodiazepine antagonist, on brainstem auditory evoked responses and on the number connection test in cirrhotic patients with subclinical neurophysiological or cognitive alterations. Thirteen cirrhotic subjects with subclinical neurophysiological or cognitive alterations were studied. A total of 3 mg of flumazenil or saline was infused intravenously. Before and after the infusion, the number connection test was administered and brainstem auditory evoked responses recorded. After 72 h, patients were crossed over. Flumazenil did not influence brainstem auditory evoked responses or the number connection test. A screening test for benzodiazepines was negative in all subjects. We conclude that benzodiazepine receptor ligands have a negligible role, if any, in the pathophysiology of subclinical neurophysiological or cognitive alterations of cirrhotic patients. PMID- 9347202 TI - The use of oral contraceptives and the occurrence of acute myocardial infarction in young women. Results from the Transnational Study on Oral Contraceptives and the Health of Young Women. AB - The objective of this study was to assess the risk of myocardial infarction (MI) associated with the use of new and old combination oral contraceptives (OC). A matched case-control study in 16 centers in Germany, the United Kingdom, France, Austria, and Switzerland explored the association of current use of combination OC with the occurrence of MI. Our subjects were 182 women aged 16-44 years with MI; the controls were 635 women without MI (at least one hospital control and one community control per case) matched for 5-year age group and region. The main outcome measures were odds ratios comparing current use of a specific group of OC against current use of other groups or against no current use. The adjusted overall odds ratio (OR; 95% confidence intervals) for MI for second generation OC versus no current use was 2.35 (1.42 to 3.89) and 0.82 (0.29 to 2.31) for third generation OC (low dose ethinyl estradiol, gestodene, and desogestrel). A direct comparison of third generation users with second generation users yielded an OR of 0.28 (0.09 to 0.86). In subgroup analyses, the odds ratio for the UK alone was 1.25 (0.36 to 4.29), while for continental Europe it was 0.10 (0.02 to 0.48). For hospital controls, the risk estimated was 0.98 (0.22 to 4.44), and 0.18 (0.04 to 0.65) for community controls. The independent risk of MI among current smokers adjusted for OC use was 7.21 (4.58 to 11.36). Among users of third generation OC, the OR for current smokers was 3.75 (0.65 to 21.74) and among users of second generation it was 9.50 (2.93 to 30.96). A comparison of OC use in the UK for the time before and after regulatory action was taken in October 1995 shows that the likelihood of a control (last control accrued June 1996) being treated with second generation OC is seven times higher after 1 November 1995 than it was before. Third generation OC are the first to be associated with no excess risk of MI. A significantly lower risk of MI is found when comparing use of third generation OC with use of second generation OC. There seems to be an impressive amelioration of risk among smokers using newer OC. An impact of regulatory action in the UK was found in the OC use spectrum of controls. PMID- 9347203 TI - First-time use of newer oral contraceptives and the risk of venous thromboembolism. AB - Recent epidemiologic studies reported that the risk of venous thromboembolism (VTE) was higher with the use of the newer third generation oral contraceptives than with second generation agents. Although the overall findings of these studies are similar, the results, as they relate to patterns and duration of oral contraceptive use particularly among first-time users, are inconsistent. We reanalyzed data from the Transnational case-control study to assess the risk of VTE associated with first-time use of oral contraceptives as a function of its duration of use. Over the period 1993 to 1995, 471 cases of venous thromboembolism were identified in Germany and the United Kingdom. For each case, up to four controls were obtained, for a total of 1772 controls. Data on oral contraceptive use and confounding variables, including data on sociodemographic, lifestyle, medical history, and family history of disease, were obtained by interview. Data analysis was based on the 105 cases and 422 controls who were first-time users of second or third generation agents, or never users of oral contraception. Rate ratios, adjusted for confounders and approximated by odds ratios, were estimated as a continuous function of duration of oral contraceptive use by logistic regression and quadratic spline models. We found, for first-time users, that the adjusted rate ratio of VTE as a function of the duration of oral contraceptive use is essentially identical for second and third generation pills relative to never users. This rate ratio increases to around 10 in the first year of use and decreases to around two after 2 years of use, remaining at this risk level thereafter for both second and third generation agents. We conclude that second and third generation agents are associated with identical risks of venous thromboembolism when they are prescribed to women who are using oral contraceptives for the first time ever. PMID- 9347204 TI - Contraceptive knowledge and attitudes of Austrian adolescents after mass media reports linking third-generation oral contraceptives with an increased risk of venous thromboembolism. AB - We performed a representative survey to determine the level of knowledge of 1,010 Austrian adolescents aged 14 to 24 years about selected facts relating to the recent massive news coverage of the increase in the risk of venous thromboembolism in users of third-generation oral contraceptives and to assess the contraceptive behavior of this population. The overall use rate of oral contraceptives and condoms had increased significantly between 1991 and 1996. Sixty-six percent of the adolescents surveyed stated not having heard or read any media reports on oral contraceptives. Only 8% of those who had knew that most reports focused on the pill as a possible cause of venous thromboembolism, whereas the majority of respondents indicated that the media conveyed doubts regarding the health safety of oral contraceptives in general. Nearly half of adolescents were unable to define what a thrombosis was. Thus, although the mass media play an important role in transmitting medical information, the dissemination of practical, accurate advice on the risks of a drug and competent patient counseling is reserved for the health care professionals. PMID- 9347205 TI - Bone mineral density during long-term treatment with Norplant implants and depot medroxyprogesterone acetate. A cross-sectional study of Thai women. AB - This cross-sectional study compares bone mineral density (BMD) in long-term Norplant implants and depot medroxyprogesterone acetate (DMPA) users. The objectives of this study were to evaluate and compare the bone mineral density between women using these contraceptives. Forty-one current users of Norplant implants and 50 DMPA users participated in the study. The BMD was measured by dual energy x-ray absorptiometry in the nondominant distal and ultradistal forearm. Serum estradiol was measured by microparticle enzyme immunoassay technique. The demographic characteristics were similar in both groups. The mean durations +/- SD of DMPA and Norplant implants were 59.14 +/- 30.73 and 31.1 +/- 11.2 months, respectively. The BMD of long-term Norplant implant and DMPA users was similar. The serum estradiol in the Norplant implant group was significantly higher than in DMPA users. However, the serum estradiol level in DMPA users ranged into normal for the follicular phase, which is higher than for postmenopausal women. This study suggests that two long-acting progestogen contraceptives do not differ with respect to their impact on BMD in long-term users. PMID- 9347206 TI - The cardiovascular safety of triphasic contraceptive steroids. AB - It was hypothesized that estrogen-induced cardioprotection is mediated by up regulation and down-regulation of expression of nitric oxide (NO) and P-selectin, respectively. Published data on circulating levels of the vasodilator NO, atherogenic glycoprotein P-selectin, and lipoprotein-a [Lp(a)] in users of triphasic contraceptive steroids are lacking. A total of 30 healthy women (nonusers, controls) and 82 women using oral triphasic contraceptive steroids (ethinyl estradiol and levonorgestrel: Triovlar, Schering AG) for 18 to 24 cycles participated in this study. Fasting blood samples were obtained from users and nonusers for the determination of P-selectin and Lp(a) by enzyme immunoassay and NO by a colorimetric method. The serum Lp(a) levels in OC users were significantly higher than those of nonusers. On the other hand, the serum NO levels in OC users were significantly elevated when compared to nonusers. Plasma P-selectin was significantly lowered in OC users p < 0.005. These results demonstrate the beneficial effects of ethinyl estradiol in the triphasic contraceptive regimen. Ethinyl estradiol may afford a degree of anti-atherogenic cardioprotective effect by up-regulation of the expression of the vasodilator NO and down-regulation of the expression of the atherogenic P-selectin. This may outweigh the cardiovascular risk of the increased atherogenic Lp(a). This study may explain the very low rate of mortality from venous thromboembolism in OC users, which compares favorably with the risks that many people accept in daily life. PMID- 9347207 TI - Effect of nonsteroidal anti-inflammatory drugs on the action of misoprostol in a regimen for early abortion. AB - Nonsteroidal anti-inflammatory drugs are frequently avoided by some investigators in protocols for medical abortion because of a concern over potential inhibition of prostaglandin action on uterine contractions. To evaluate the effects of nonsteroidal anti-inflammatory drugs on uterine cramping and resultant pregnancy expulsion, analgesic use by 449 participants with gestational age of < or = 56 days in three previously reported medical abortion trials involving methotrexate and misoprostol was reviewed. Subjects from each of the three trials were included in this analysis only if they received 50 mg/m2 methotrexate intramuscularly followed 3 or 7 days later by 800 micrograms misoprostol vaginally. Misoprostol administration was repeated if the abortion did not occur after the first misoprostol dose. A total of 416 subjects met the study criteria. For the women who took a nonsteroidal anti-inflammatory drug after the first dose of misoprostol, 132/246 (53.7%) aborted within 24 h. For those that did not take a nonsteroidal anti-inflammatory drug, 83/170 (48.8%, p = 0.38) aborted within 24 h. Similarly, 27/56 (48.2%) of the women who took a nonsteroidal anti inflammatory drug aborted within 24 h of the second dose of misoprostol. However, only 32/145 (22.1%, p = 0.002) of the women who did not take a nonsteroidal anti inflammatory drug aborted. Use of a nonsteroidal anti-inflammatory drug does not interfere with the action of misoprostol to induce uterine contractions and pregnancy expulsion in women receiving methotrexate and misoprostol for early abortion. PMID- 9347208 TI - Misoprostol 3, 4, or 5 days after methotrexate for early abortion. A randomized trial. AB - A randomized trial was conducted including 287 pregnant women seeking elective abortion to compare the efficacy of misoprostol given 3, 4, or 5 days after methotrexate for abortion at < or = 63 days' gestation. Subjects received 50 mg/m2 methotrexate intramuscularly and were randomly allocated to self-administer vaginally 800 micrograms of misoprostol 3, 4, or 5 days after the methotrexate. The misoprostol dose was repeated 48 and 96 h later if the abortion did not occur. Outcome measures included successful abortion (complete abortion without requiring a surgical procedure), and side effects. Eighty-six cases (93%; 95% confidence interval [CI] 85%-97%) aborted in Group I; 90 cases (92%; 95% CI 84% 96%) aborted in Group II (relative risk [RR] = 1.09; RR 95% CI 0.38-3.14); and 89 (93%; 95% CI 86%-97%) cases aborted in Group III (RR = 0.97; RR 95% CI 0.33 2.87). No significant statistical differences were obtained for the success rates when misoprostol was given days 3, 4, or 5 after the administration of methotrexate (p = 0.97) nor with any of the characteristics of the subjects. Complete abortion occurred in 265/287 (92%; 95% CI 89%-95%) patients. Twenty-two cases (8%; 95% CI 5%-11%) resulted in failure. Side effects for methotrexate were minimal while for misoprostol they were moderate. This combination could be an alternative to surgical abortion or the use of antiprogestins and prostaglandins for medical abortion. PMID- 9347209 TI - Effect of once weekly administration of mifepristone on ovarian function in normal women. AB - The effects of mifepristone on ovarian function during a once weekly oral administration regimen were studied in nine healthy women. Each received 25 mg mifepristone on cycle days 3, 10, 17, and 24. Ovulation, as documented by hormonal measurements and ultrasonography, was inhibited during treatment in five subjects, with a midcycle surge of luteinizing hormone and ovulation occurring 6 18 days after the last pill was administered in four of the five subjects. These five treatment cycles were prolonged 9-26 days. The other four subjects had normal cycles as judged by serum hormone levels, ultrasonography, and cycle length. All nine subjects had delayed endometrial growth as indicated by ultrasonography. There was a significant correlation between concentrations of serum mifepristone (10 h and 58 h) and alpha 1-acid glycoprotein, the protein to which mifepristone binds in circulation. Response to mifepristone did not depend on its circulating levels. We conclude that once weekly administration of 25 mg mifepristone can interfere with normal follicular development and function, but the inhibition of ovulation was inconsistent. PMID- 9347210 TI - Short-term treatment with levonorgestrel does not antagonize the ethinyl estradiol-induced increase of uterine blood flow in postmenopausal women. AB - The aim of this study was to determine the effect of a short-term ethinyl estradiol/levonorgestrel medication on blood flow in the uterine arteries in postmenopausal women in a prospective placebo-controlled double-blind study. Twenty-one healthy postmenopausal woman at least 2 years after menopause received 60 micrograms ethinyl estradiol (EE) for 14 days followed by 40 micrograms EE plus 125 micrograms levonorgestrel (LNG) for 12 days (total treatment period 26 days). Sonographically, uterine volume, endometrial thickness, and blood flow in the uterine arteries [as reflected by pulsatility (PI) and resistance indices (RI)] were measured. Uterine size increased from 44 to 80 mL (day 14, p < 0.001) and 87 mL (day 26, p = NS). Endometrium grew from 3 to 8 mm (day 14, p < 0.001) and 11 mm (day 26, p = NS). Uterine arterial PI fell from 2.76 to 1.37 (day 14, p < 0.001) and 1.34 (day 26, p = NS), whereas RI fell from 0.9 to 0.68 (day 14 and day 26, p < 0.001). In conclusion, short-term treatment with LNG does not antagonize the vascular effect of EE on the uterine arteries as reflected by PI and RI. This result might have clinical significance in the selection of the progestin used in hormonal replacement therapy. PMID- 9347211 TI - Modulation of ovarian function by an oral contraceptive containing 30 micrograms ethinyl estradiol in combination with 2.00 mg dienogest. AB - Twenty-two healthy female volunteers with normal ovulatory cycles, aged between 20 and 34 years (27.3 +/- 4.1), were included in a single-center, noncomparative study to investigate the modulation of ovarian function by an oral contraceptive containing 30 micrograms ethinyl estradiol in combination with 2.00 mg dienogest. At baseline, during three treatment cycles and post-treatment, serum levels of luteinizing hormone, follicle-stimulating hormone, 17 beta-estradiol, and progesterone were assayed and ultrasonography was used to measure follicular size and the thickness of the endometrium. The primary efficacy variable was inhibition of ovulation as measured by ovarian activity grading. All volunteers ovulated during the pretreatment cycle. During treatment, none of the subjects had ovulatory cycles, although there was still some ovarian activity in several subjects. During the first treatment cycle, only 4% (1 subject) of cycles showed active follicle-like structures. The frequency of follicle-like structures increased to 33% and 35% during treatment cycles 2 and 3. The frequency of presumptive luteinized unruptured follicle-like structures was 5% (1 subject) and 15% (3 subjects) in treatment cycles 2 and 3. The serum hormone concentrations were effectively suppressed in comparison to baseline. The ovarian activity returned to baseline during the post-treatment period. One subject was excluded from further study because of a medical problem believed unrelated to use of the oral contraceptive. No serious adverse events were recorded during the course of the study. The results of the present investigation indicate that the modulatory effects on ovarian function of the monophasic oral-contraceptive containing 30 micrograms ethinyl estradiol combined with 2.00 mg dienogest lead to adequate suppression of ovarian activity and effective inhibition of ovulation. PMID- 9347212 TI - Markers of risk after acute myocardial infarction. A comparison of clinical variables, ambulatory and exercise electrocardiography, echocardiography, and stress echocardiography. AB - BACKGROUND: Short-term mortality after myocardial infarction has decreased continuously among members of selected populations. Nonetheless, the long-term prognosis among members of unselected populations remains bad. Further research in risk stratification is therefore needed. In the present study we tested the additive value of clinical variables, echocadiography, ambulatory electrocardiography, exercise testing, and stress echocardiography in assessing the long-term prognosis after acute myocardial infarction. METHODS: Two dimensional echocardiography and ambulatory electrocardiography (analysis of ST segment changes and of heart rate variability) were performed for 74 patients aged < 75 years who had had an acute myocardial infarction. Before their discharge from hospital, 70 patients were subjected to a combined exercise test and stress echocardiography. The time of follow-up was > or = 3 years. RESULTS: During follow-up 18 patients died, and 38 suffered cardiac events defined as death, nonfatal reinfarction and the need for revascularization. We first tested 31 covariates in a univariate regression analysis. A subsequent multivariate analysis was performed in two stages. During the first of these, clinical variables (a history of systemic hypertension, infarct localization, and diabetes mellitus) and variables derived from noninvasive tests (new-onset wall-motion abnormality during stress echocardiography, ST-segment depression and heart-rate variability during ambulatory electrocardiography, the ejection fraction by echocardiography at rest, and the double product during exercise tests) predicted mortality. After the second stage, however, the only remaining independent predictors of mortality were the presence of a new-onset wall-motion abnormality (P < 0.0001, relative risk 13.5, 95% confidence interval 3.6-51.3), ST-segment depression during ambulatory electrocardiography (P = 0.003, relative risk 5.0, 95% confidence interval 1.7-15.7) and a decreased heart rate variability (P = 0.007). CONCLUSIONS: The only variables that were of independent value in assessing the long-term mortality were those expressing residual myocardial ischemia and the cardiovascular sympatho-vagal balance. It is, therefore, recommended that one should monitor these variables for patients recovering from an acute myocardial infarction. PMID- 9347213 TI - Myocardial and gastrointestinal release of vasoactive intestinal peptide during experimental acute myocardial infarction. AB - BACKGROUND: Vasoactive intestinal peptide (VIP) acts as a vasodilator on coronary and gastrointestinal arteries. During coronary occlusion, the locally released VIP may exert a protective effect on the heart, but it may aggravate the shock state through its vasodilatory effect in the gastrointestinal tract. METHODS: After left thoracotomy, the left circumflex coronary artery (LCx) was prepared, and a pneumatic occluder was introduced around it. After 60 min of coronary occlusion, the LCx was reperfused in six dogs (reperfusion group), while in another six the occlusion was maintained for 6 h (occlusion group). Five dogs served as sham-operated controls. The plasma concentration of VIP was determined at baseline, after the 60 min occlusion and 10 min, 3 h and 6 h after reperfusion, or 3 h and 6 h after continuous occlusion in the coronary sinus and in the femoral and portal veins. RESULTS: The plasma VIP concentrations in all three vessels were increased after 60 min of LCx occlusion. During the 6 h constant coronary occlusion, concentrations remained increased in both the coronary sinus and the portal vein, but not in the femoral vein. In the reperfusion group, 10 min after reperfusion, the plasma concentrations of VIP in all three vessels had decreased. CONCLUSIONS: Coronary artery occlusion causes a long-term increase in plasma VIP concentrations that decreases after reperfusion, when measured in the portal vein and coronary sinus, but not in the femoral veins. PMID- 9347214 TI - Role of the angiotensin II type 1 receptor in preconditioning against infarction. AB - OBJECTIVE: To assess the role of angiotensin II type 1 (AT1) receptor in the mechanism of limitation of infarct size by preconditioning. METHODS: Myocardial infarction was induced by 30 min coronary artery occlusion and 3 h reperfusion in the rabbit. The infarct size was determined by tetrazolium staining and expressed as a percentage of the area at risk (%ISAR). Rabbits were subjected to one of the following four treatments: no drug (i.e. control); administration of 1 mg/kg CV 11,974, a specific AT1 receptor antagonist, 20 min before ischemia; preconditioning with 5 min ischemia and then 5 min reperfusion; and administration of CV-11,974 plus preconditioning. RESULTS: It was confirmed that the same dose of CV-11,974 (i.e. 1 mg/kg) could block the pressor response to injection of 0.5 microgram/kg angiotensin II completely for 60 min. The %ISAR for the untreated control group was 45.1 +/- 3.9%, and administration of CV-11,974 alone did not modify the %ISAR (47.5 +/- 5.9%). The %ISAR for the group administered both CV-11,974 and subjected to preconditioning (30.4 +/- 3.5%) was modestly smaller than that for the controls, but was significantly larger than for the preconditioned group (12.0 +/- 1.8%). The angiotensin II level in arterial plasma did not differ before and after the 5 min preconditioning. It was shown with separate groups of rabbits that a modest protection conferred by preconditioning with 3 min ischemia was also attenuated by administration of CV 11,974 (%ISAR 28.4 +/- 4.3 with CV-11,974 versus 19.3 +/- 1.7% without CV-11,974, P < 0.05). CONCLUSION: These results suggest that activation of AT1 receptors by angiotensin II produced locally in the heart contributes to the limitation of infarct size by preconditioning. PMID- 9347216 TI - Simpson's rule for the volumetric ultrasound assessment of atherosclerotic coronary arteries: a study with ECG-gated three-dimensional intravascular ultrasound. AB - BACKGROUND: Volumetric intravascular ultrasound (IVUS) assessment provides complementary information on atherosclerotic plaques. The volumes can be calculated by applying Simpson's rule to cross-sectional area data of multiple IVUS images, acquired with a fixed sample spacing, which is the distance (along the vessel's axis) between two images. OBJECTIVE: To evaluate the effect of different sample spacings on the results of volumetric IVUS measurements. METHODS: A stepwise electrocardiographically gated IVUS image-acquisition and automated three-dimensional analysis approach was applied to 26 patients. Twenty eight coronary segments with mild-to-moderate coronary atherosclerosis were examined. Volumetric measurements of five images per mm (i.e. sample spacing 0.2 mm), representing a complete scanning of the coronary segment, were considered the optimal standard, against which volumetric measurements of three, one, and one-half images per mm (i.e. larger sample spacings) were compared. RESULTS: The lumen, total vessel, and plaque volumes obtained with five images per mm were 183.3 +/- 2.8, 350.6 +/- 141.6, and 167.3 +/- 89.2 mm3. There was an excellent correlation (r = 0.99, P < 0.001) between these data and volumetric measurements with larger sample spacings. The volumetric measurements with larger sample spacings differed on average only by a little (< 0.7%) from the optimal standard measurements. However, a relatively small, but significant, increase in SD of these differences was associated with the wider sample spacings (< 3.6%, P < 0.05). CONCLUSIONS: The width of the sample spacing has a relatively small but significant impact on the variability of volumetric intravascular ultrasound measurements. This should be considered when designing future volumetric studies. The electrocardiographically gated acquisition of five IVUS images per mm axial length during a stepwise transducer pull-back is an ideal approach, particularly when addressing with IVUS volumetric changes that are assumed small, such as those expected in studies of the progression and regression of atherosclerosis. PMID- 9347215 TI - Acute effects of 17 beta-estradiol on the coronary microcirculation: observations in sedated, closed-chest domestic swine. AB - OBJECTIVES: To test the hypotheses: that acute administration of 17 beta estradiol dilates normal coronary microvessels in vivo; that coronary microvascular responses to acute estrogen stimulation exhibit sexual dimorphism; and that nitric oxide has a role in mediating these effects. METHODS: Measurements of hemodynamics, coronary flow velocity (Doppler), myocardial blood flow (microspheres) and oxygen consumption were made in closed-chest swine: group 1 consisted of castrated juvenile males, groups 2 and 3 of estrogen pretreated, castrated juvenile males, and group 4 of sexually mature females. 17 beta Estradiol (2, 20 or 200 ng/kg) was given by intracoronary injection and data obtained 20-30 min later; additional measurements were made 1 h after the 200 ng/kg dose. The effect of L-NG-monomethylarginine (L-NMMA) on 17 beta-estradiol responses was also tested. Tissue and blood concentrations of 17 beta-estradiol, and concentrations of estrogen receptor in myocardium and coronary vessels were obtained. RESULTS: In estrogen-naive castrated males, 17 beta-estradiol had no effect on coronary flow velocity or myocardial blood flow, but 1 h after the 200 ng/kg dose there was an increase in diastolic coronary resistance compared with baseline (48 +/- 20 versus 41 +/- 17 mmHg/mkHz; P < 0.05). Estrogen pretreated castrated males also showed no change in myocardial blood flow after 17 beta estradiol, but coronary flow velocity decreased (P < 0.05) compared with baseline 1 h after the 200 ng/kg dose (from 1.69 +/- 0.61 to 1.41 +/- 0.42 kHz) and diastolic coronary resistance increased significantly (P < 0.01) compared with control at this time (51 +/- 15 compared with 39 +/- 14 mmHg/mkHz). In sexually mature females, 17 beta-estradiol had no effect on myocardial blood flow but did cause a significant (P < 0.05) decrease in diastolic coronary vascular resistance compared with baseline (51 +/- 9 mmHg/mkHz) at both the 20 ng/kg and the 200 ng/kg doses (both 43 +/- 11 mmHg/mkHz). Coronary flow velocity also increased (P < 0.06) compared with baseline (1.34 +/- 0.26 mmHg/mkHz) after the 200 ng/kg dose (1.69 +/- 0.61 mmHg/mkHz). L-NMMA had no effect on flow responses to 17 beta estradiol in any group. Classical estrogen receptors were not present in myocardium or coronary arteries from male or female swine. CONCLUSIONS: These results demonstrate that 17 beta-estradiol exerts a mild constrictor effect on the coronary microvessels of normal castrated, juvenile males whether estrogen naive or estrogen-pretreated. In contrast, sexually mature normal females exhibit mild dilatation of the coronary microcirculation in response to acute estrogen stimulation. Nitric oxide does not appear to have a role in mediating the dilator response in females, and classical estrogen receptors are not involved. A direct membrane effect of the hormone (perhaps via alteration in potassium conductance) seems likely, and demonstrates sexual dimorphism. PMID- 9347217 TI - Extracorporeal membrane oxygenation for cardiac support. AB - Extracorporeal membrane oxygenation (ECMO) uses cardiopulmonary bypass technology to provide prolonged cardiac or respiratory support in the intensive care unit. The use of ECMO for neonatal respiratory failure is now good evidence-based medicine following publication of the UK Collaborative ECMO Trial, but its use in adults and children remains controversial. In this review the use of ECMO to support paediatric patients with pre- and post-operative cardiac insufficiency is discussed. The survival with ECMO in these patients is 43-61%, which is remarkable in a group of patients who are moribund prior to initiation of ECMO. PMID- 9347218 TI - The role of nitric oxide and cytokines in heart failure. AB - Heart failure is a common problem associated with considerable mortality and morbidity. The mechanisms underlying the heart failure syndrome, which remain poorly understood, may involve an inflammatory process. Nitric oxide (NO) and various cytokines could play an important role in this inflammatory process. Recent evidence has emerged in both animal models and humans suggesting that both of these mediators may play an important role in heart failure. NO is synthesized by the NO synthase family of enzymes. Two of these enzymes are constitutive, endothelial NO synthase and neuronal NO synthase. The third enzyme, inducible NO synthase, is capable of producing large amounts of NO once induced by mediators such as interleukin (IL)-1, IL-2, IL-6, tumour necrosis factor (TNF)-alpha, and interferon-gamma. Endothelial NO synthase is present in the heart in the endocardium, cardiac myocytes, and cardiac conduction tissue. Inducible NO synthase is present in cardiac myocytes, endocardium, vascular smooth muscle cells, and infiltrating inflammatory cells. Evidence from both animal models and patients suggests that NO exerts a negative inotropic effect. Increased inducible NO synthase, TNF-alpha, and IL-6 have been found in patients with heart failure in several studies. In other studies, decreased endothelial NO synthase was found in patients with heart failure. TNF-alpha and IL-6 may be produced in heart failure and may induce inducible NO synthase, resulting in NO production, which acts as a negative inotrope. Endothelial NO synthase may be decreased as a result of downregulation by TNF-alpha or inducible NO synthase. The possible role of these mediators in heart failure needs further evaluation because these findings could have novel therapeutic implications. PMID- 9347219 TI - Bibliography current world literature. PMID- 9347220 TI - Programmed cell death in prokaryotes. AB - Programmed cell death (PCD), also referred to as apoptosis, is a cellular "suicide" mechanism, based on information from its own internal metabolism, environment, developmental history, and genome. This system was described in eukaryotes continuously along evolution, through amoebae, nematodes, insects, and animals. PCD is essential for the proper development or function of a cell system, organ, or survival of the organism as a whole. Research in the last 2 decades has shown that the life cycle of several prokaryotic organisms display developmental programs, similar to metazoan differentiation, that is part of their adaptation to stressful environments. These include warmer cell formation and differentiation in Caulobacter cereus, sporulation in Bacillus and Streptomyces, heterocyst formation in Anabaena, development of bacteroids in Rhizobium, the formation of multicellular fruiting bodies and sporulation in Myxobacteria, and the formation of nonculturable, but viable, cells in various Gram-negative bacteria. Moreover, and more significantly, the photosynthetic bacteria Rhodobacter capsulatus were shown to release nucleoprotein particles designated "gene transfer agent (GTA)" as they enter the stationary phase. GTAs contain DNA of 3.6 x 10(6) molecular weight, representing all parts of the genome, and they may be taken up by other strains of R. capsulatus, and complement mutants. We postulate that these various modes of stress adaptations in bacteria are prokaryotic manifestation, and possibly the phylogenetic precursor, of the eukaryotic phenomenon, programmed cell death, and therefore we propose to designate it "proapoptosis". In addition to their function, apoptosis and proapoptosis share various mechanistic programmed features, including DNA fragmentation and packaging, cell shrinkage, degradation of RNA, proteolysis and synthesis of new proteins, and the involvement of reactive oxygen species. PMID- 9347221 TI - Aspects of Toxocara epidemiology: human toxocarosis. AB - Toxocarosis is the clinical disease in man caused by infection of zoonotic roundworms of dogs and cats, Toxocara canis and T. cati. In this review the mode of transmission to the human by oral ingestion of Toxocara eggs from the environment is discussed. T. cati seems to play a more important role than generally suggested. Direct contact with animals is not considered a potential risk because embryonation of excreted Toxocara ova requires a minimum of 2 weeks. For the same reason there is no relationship expected between infection and exposure to dogs and cats in the household. Children more frequently have clinical symptoms because of the closer contact with contaminated soil in yards and sandpits, the lack of hygiene, and because of eating dirt. Toxocara larval migration in the body can cause various clinical syndromes. Visceral larva migrans, ocular larva migrans, and covert toxocarosis are described. Serodiagnostic techniques are reliable tools to detect antibodies or antigens. Systemic treatment with anthelmintics is described but can result in hypersensitivity reactions caused by dying larvae. For ocular lesions, laser photocoagulation and corticosteroid therapy are described. Preventive measures consist of preventing contamination of the environment with Toxocara eggs and for education of pet owners and non-pet owners to increase awareness about potential zoonotic hazards. Veterinary practitioners, general practitioners, and public health agencies should therefore provide sufficient information and advice. PMID- 9347222 TI - Aspects of Toxocara epidemiology: toxocarosis in dogs and cats. AB - Toxocara canis and Toxocara cati are common roundworms of dogs and cats. In this review the life cycles of these parasites are described, including the various routes of transmission, such as transplacental, transmammary infection, and infection through paratenic hosts. The somatic and tracheal migration in the body of the hosts after infection with Toxocara eggs or larvae is discussed, with special reference to age resistance and differences between dog and cat. The clinical symptoms and pathology in adult and young dogs and cats are given. Diagnosis of patent infections can be obtained by fecal examination, and treatment consists of the use of anthelmintics. Control of the infection and disease is achieved by prevention of contamination of the environment, anthelmintic treatment strategy, and education. Special attention is given to the efficacy of anthelmintics against adult worms and against somatic larvae. It is concluded that education on the life cycles of the parasites, hygiene, and anthelmintic treatment schedules is required because of the zoonotic risks of Toxocara spp. Deworming of pregnant dogs and cats is not recommended. PMID- 9347223 TI - Leishmania phagolysosome: drug trafficking and protein sorting across the compartment. AB - Survival or destruction of intramacrophage pathogen Leishmania depends in part on modulation of their host cell phagosome, capabilities of the infected macrophages to present parasite antigen to the host's immune system. Macrophages house these parasites as amastigotes in the acidic phagolysosomal compartment. Leishmania phagolysosome is the potential site for processing and presentation of its antigen as well as being the target site for chemotherapy in leishmaniasis. It is thought that the parasites are killed from macrophage activation by lymphokines secreted from either helper T1 cells or CD8+ T cells. Characterization of both the host and parasite molecules in the compartment in the context of biogenesis of Leishmania-phagolysosome and processing of the parasite antigen by this compartment are discussed. Trafficking of different drugs and new agents through this compartment and their role in chemotherapy and necessity of developing new drug carrier are also stressed. PMID- 9347224 TI - Toxicity and carcinogenicity of Cr(VI) in animal models and humans. AB - The toxicity and carcinogenicity of hexavalent chromium (Cr) in animal and human models are reviewed. The focus of this review is not on the well-established fact that hexavalent Cr compounds of low and high water solubility can induce respiratory cancers, but rather this review addresses other types of cancers induced by exposure to hexavalent Cr compounds. Additionally, non-cancer endpoints are also discussed with documentation of human and animal studies showing non-cancer health effects of hexavalent Cr exposure on the respiratory system, GI system, immune system, liver, and kidney. There is an emerging understanding that because hexavalent chromate is isostructural with phosphate and sulfate, it is readily taken up by the G.I. tract and penetrates to many tissues and organs throughout the body. This is supported by animal studies and experiments using human volunteers. From the epidemiological studies, there is suggestive evidence that hexavalent Cr causes increased risk of bone, prostate, lymphomas, Hodgkins, leukemia, stomach, genital, renal, and bladder cancer, reflecting the ability of hexavalent chromate to penetrate all tissues in the body. A high accumulation of Cr(III) in all tissues and organs is a strong indication of the wide toxic potential of exposure to soluble hexavalent Cr in the drinking water and in the ambient environment. PMID- 9347225 TI - Hematopoietic and lymphatic malignancies in vehicle mechanics. AB - Although it is generally acknowledged that benzene causes leukemia, especially acute myeloid leukemia, considerable divergences persist in the assessment of the leukemia risk due to occupational low-level benzene exposure. Specifically, the risk for vehicle mechanics is considered by some authors as being nondetectable with epidemiologic methods, whereas others calculated that the incidence rate of leukemia (all types) in vehicle mechanics is increased more than 60 times. The purpose of this review is to examine the publications on this topic in light of criteria for causal inference and to discuss the possible role of bias, confounding factors, and chance. The results of this analysis reveal that there are surprisingly few epidemiologic observations supporting an increased incidence of leukemia in vehicle mechanics. Apparently, publications suggesting a leukemogenic effect of low-level benzene exposure in garage mechanics are more often quoted than their negative counterparts, although they are not better designed. PMID- 9347226 TI - Toxic effects of griseofulvin: disease models, mechanisms, and risk assessment. AB - Griseofulvin (GF) has been in use for more than 30 years as a pharmaceutical drug in humans for the treatment of dermatomycoses. Animal studies give clear evidence that it causes a variety of acute and chronic toxic effects, including liver and thyroid cancer in rodents, abnormal germ cell maturation, teratogenicity, and embroyotoxicity in various species. No sufficient data from human studies are available at present to exclude a risk in humans: therefore, attempts were made to elucidate the mechanisms responsible for the toxic effects of GF and to address the question whether such effects might occur in humans undergoing GF therapy. It is well documented that GF acts as a spindle poison and its reproductive toxicity as well as the induction of numerical chromosome aberrations and of micronuclei in somatic cells possibly may result from disturbance of microtubuli formation. Likewise, a causal relationship between aneuploidy and cancer has been repeatedly postulated. However, a critical survey of the data available on aneuploidogenic chemicals revealed insufficient evidence for such an association. Conceivably, other mechanisms may be responsible for the carcinogenic effects of the drug. The induction of thyroid tumors in rats by GF is apparently a consequence of the decrease of thyroxin levels and it is unlikely that such effects occur in GF-exposed humans. The appearance of hepatocellular carcinomas (HCC) in mice on GF-supplemented diet is preceded by various biochemical and morphological changes in the liver. Among these, hepatic porphyria is prominent, it may result from inhibition of ferrochelatase and (compensatory) induction of ALA synthetase. GF-induced accumulation of porphyrins in mouse liver is followed by cell damage and necrotic and inflammatory processes. Similar changes are known from certain human porphyrias which are also associated with an increased risk for HCC. However, the porphyrogenic effect of GF therapy in humans is moderate compared with that in the mouse model, although more detailed studies should be performed in order to clarify this relationship on a quantitative basis. A further important effect of GF-feeding in mice is the formation of Mallory bodies (MBs) in hepatocytes. These cytoskeletal abnormalities occur also in humans, although under different conditions; their appearance is associated with the induction of liver disease and HCC. Chronic liver damage associated with porphyria and MB formation, enhanced cell proliferation, liver enlargement, and enzyme induction all may contribute to the hepatocarcinogenic effect of GF in mice. In conclusion, further investigation is required for adequate assessment of health risks to humans under GF therapy. PMID- 9347227 TI - Dental mercury amalgam: Part I. The nature of mercury amalgam. PMID- 9347228 TI - Congenital neurocutaneous melanosis. PMID- 9347229 TI - Bits and pieces. PMID- 9347230 TI - Eruptive milia. AB - Milia are small subepidermal keratin cysts. They may arise either spontaneously or in the course of bullous diseases or trauma. We describe an unusual case of multiple eruptive milia on the posterior neck and back and review the classification of milia. PMID- 9347231 TI - Autoimmune idiopathic thrombocytopenic purpura with the subsequent occurrence of systemic lupus erythematosus. AB - Cutaneous manifestations of petechiae, purpura, and ecchymosis can lead the physician to discover an underlying platelet abnormality. Autoimmune idiopathic thrombocytopenic purpura (AITP) is a diagnosis of exclusion, mediated by a destructive IgG antibody response to the platelets' membrane components. In addition to showing evidence of cutaneous and mucosal bleeding (ie, epistaxis, hematuria), patients with AITP are at an increased risk for systemic lupus erythematosus (SLE). Therefore, it is suggested that patients with AITP be closely monitored for SLE. PMID- 9347232 TI - Congenital insensitivity to pain with anhidrosis. AB - Congenital insensitivity to pain with anhidrosis is one of a group of rare diseases termed hereditary sensory-motor neuropathies. Primary clinical features of this entity include congenital analgesia, inability to sweat, and mental retardation. Besides the rarity of these clinical entities, difficulty in evaluating the sensory disturbances, especially in small children, makes the diagnosis a clinical problem. In this article a 3-year-old boy, with consanguineous parents and no family history of the disorder, who was evaluated for two years because of ulcerating lesions on his knees, is presented. Physical examination revealed deep ulceration on his knees and scars from burns on his neck and scalp. Moderate mental retardation and analgesia were noted. There was symmetrical loss of pain and touch sensation on his hands and feet. Electromyographic examination showed absence of action potentials of the ulnar and sural nerves, decrease in the sensory and motor nerve conduction velocities, and amplitude of action potentials. The result of the application of pilocarpine showed anhidrosis. His skin and nerve biopsy specimens were also examined. PMID- 9347234 TI - Facial scarification and tattooing on Santa Catalina Island (Solomon Islands). AB - Ritual scarification is the culturally sanctioned process of incising the skin to achieve patterned scars. Scarification was practiced widely by traditional societies, but the encroachment of Western cultural expectations has made the practice increasingly uncommon. Ritual tattooing has a meaningful place in many traditional societies. Ritual scarification and tattooing are still found on Santa Catalina Island, an isolated member of the Solomon Islands in the south west Pacific. PMID- 9347233 TI - Clinical cure of fungal madura foot with oral itraconazole. AB - We report the case of a 26-year-old Malian patient who presented with mycetoma of the foot and frank bone involvement caused by Madurella mycetomatis. Long-term itraconazole therapy was clinically effective and well tolerated. PMID- 9347235 TI - Cardioversion dermatitis. AB - Dermatitis resulting from medical devices is common. The dermatologic hazards of transcutaneous electrical nerve stimulation, cardiac pacemakers, and electrocardiography electrodes are well described. We report a 70-year-old man who experienced contact dermatitis from direct current cardioversion. To our knowledge, this is the first report of cardioversion-associated dermatitis. PMID- 9347236 TI - Tuberculoma of the tongue presenting as macroglossia. AB - Lingual tuberculosis presenting as macroglossia in a 20-year-old woman has been reported. The histopathologic picture was similar to lupus vulgaris and the intradermal tuberculin test showed a strongly positive vesiculobullous reaction after forty-eight hours. No active internal focus of tubercular infection could be detected. The patient responded well to antituberculous chemotherapy. The relative lack of symptoms suggest that the enlargement had been a slow process resulting from the lodging of bacilli in the tongue. PMID- 9347237 TI - Rheumatoid neutrophilic dermatitis. AB - Rheumatoid neutrophilic dermatitis (RND) is a rare cutaneous finding in patients with severe rheumatoid arthritis or with high-titer rheumatoid factors. Most commonly, these lesions are erythematous papules or plaques distributed symmetrically on extensor surfaces. On histologic examination a dense dermal neutrophilic infiltrate without vasculitis is apparent. The pathogenesis of RND is unclear, and few treatments are known. With careful clinical and histologic examination, RND may be differentiated from the wide array of other cutaneous findings in rheumatoid arthritis. PMID- 9347238 TI - Anaplastic large cell lymphoma mimicking noduloulcerative tertiary syphilis. AB - Anaplastic large cell lymphoma (ALCL) is a recently described malignancy that can show heterogeneity in its clinical presentation. We report on a patient whose clinical presentation, medical history, and serologic test results were suggestive of tertiary syphilis. Histologic evaluation, however, revealed a large cell lymphoma with Ki-1 positive findings on immunostaining. We review the differential diagnosis of ALCL and add to that a pattern of ALCL clinically mimicking noduloulcerative tertiary syphilis. PMID- 9347239 TI - Services for children with disabilities in European countries. PMID- 9347240 TI - Impaired insulin but normal pentagastrin effect on gastric acid secretion in diabetic rats: a role for nitric oxide. AB - Significant changes in gastrointestinal function, decreased gastric secretion and motility in particular, are often observed in patients with chronic diabetes. The mechanisms leading to those remain unclear. In these studies we evaluated the gastric acid secretory response to insulin and pentagastrin in normal Wistar and streptozotocin diabetic rats. We also sought to determine the role of nitric oxide (NO) in this process. The animals were anesthetized with sodium pentobarbital. Warm saline was perfused through a polyethylene tube placed in the oesophagus and collected from the duodenum at 10 min intervals. Following a 50 min equilibration period, a bolus intra-jugular infusion of insulin (4.0 U/kg), 2 deoxyglucose (200 mg/kg) or pentagastrin 4.0 (ug/kg) was started and samples of the gastrointestinal perfusate were collected for an additional 80 min. Insulin stimulated acid secretion peaked 60 min after bolus infusion in normal animals; a response that was significantly decreased in the diabetic rats. Similarly, 2 deoxyglucose-induced glucopenia increased gastric acid secretion to a lower extent in diabetic versus normal rats. The stimulatory response to pentagastrin was prompt and essentially equal in normal and diabetic animals. However, when hypoglycemia was prevented by glucose infusion, insulin did not stimulate gastric acid secretion in normal rats. Further, glucose infusion in these animals actually enhanced the secretory response to pentagastrin. Nitro-L-arginine methyl ester (L-NAME 20 mg/kg i.v.), an inhibitor of NO synthetase, also prevented the secretory response to insulin but not to pentagastrin. Preinfusion of arginine (100 mg/kg i.v.) in diabetic rats restored the gastric secretory response to insulin toward that of normal animals. We conclude that the gastric acid secretory response to insulin, but not to pentagastrin, is decreased in diabetic animals, that this response may operate through a NO mediated mechanism possibly set in motion by central nervous system glucopenia and that this NO-mediated mechanism is attenuated in diabetes. PMID- 9347241 TI - The endocrine pancreas in virgin and pregnant offspring of diabetic pregnant rats. AB - Diabetes of the mother during pregnancy induces structural and functional adaptations in the fetal endocrine pancreas. We have previously shown in our experimental rat model, that the impact of this abnormal intra-uterine milieu leads, in the adult offspring, to a disturbance of the glucose homeostasis and to the development of gestational diabetes. The aim of the present work is to investigate wether these functional differences can be explained by structural differences at the level of the endocrine pancreas. Therefore the size and the structure of the endocrine pancreas, as well as the contribution of the insulin-, glucagon-, somatostatin- and PP-cells, were investigated morphometrically in the adult youngsters of mildly and of severely diabetic mothers, since both display a disturbed glucose tolerance but with divergent characteristics. Also the adaptation of their endocrine pancreas to pregnancy was measured and compared to that of a control pregnancy. In the offspring of mildly diabetic mothers, the size of the endocrine pancreas and the distribution of the islets of Langerhans are normal. Also the doubling of the endocrine mass during pregnancy is similar to controls. The high proportion of A-cells, especially in relation to a normal B cell mass and the low amount of PP-cells, might play a role in the impairment of the insulin response in these animals and in the development of gestational diabetes. In the offspring of severely diabetic mothers a clear hypertrophy of the endocrine pancreas is noted, which is mainly due to the presence of numerous small islets and which does not increase further during pregnancy. In these animals, the size of the endocrine pancreas and of the B-cell mass have reached 'pregnant' values without pregnancy, which coincides with an exaggerated insulin output and peripheral insulin resistance, as during normal pregnancy. No further increase in islet mass is seen during pregnancy, which is associated with gestational diabetes. PMID- 9347242 TI - Poor vision as a contributory factor in diabetic neuro-arthropathy. AB - Two patients with longstanding insulin-dependent diabetes mellitus (IDDM) complicated by neuropathy, nephropathy and retinopathy leading to poor vision each developed a swollen and relatively painless foot. The previous day, both had been walking over uneven ground. At initial presentation, no X-ray was taken of the foot in case one and no action was taken over an undisplaced navicular fracture in case two. A few weeks later, a midfoot Charcot had developed in both cases. Poor vision may increase the risk of injury to the feet and may be an important risk factor for Charcot neuro-arthropathy. Plain radiographs are an important investigation for the swollen insensitive foot, even in the absence of pain. PMID- 9347243 TI - Glycosaminoglycan sulodexide decreases albuminuria in diabetic patients. AB - Albuminuria is a dominant biochemical feature of developing diabetic nephropathy. A disturbed metabolism of heparan sulphate characterized by an increased loss of anionic charges in the basement membrane has been considered as one of the main factors causing an increased albumin output into urine. All therapeutic approaches inducing a reduction of the albumin excretion rate (AER) have a protective effect on renal function. The effect of glycosaminoglycan sulodexide on albuminuria was studied in a group of 53 diabetic patients (26 Type 1 and 27 Type 2) with micro and macroalbuminuria. Sulodexide (Vessel Due F) was administered intramuscularly in one daily dose (600 lipasemic units) for 3 weeks followed by a 6 week wash-out period. A significant decrease of AER was found in a total cohort of patients following just 1 week of sulodexide treatment (mean 162 micrograms/min, range 10-2708 micrograms/min vs mean 248 micrograms/min, range 20-3160 micrograms/min, P < 0.001). This effect lasted 3-6 weeks after drug withdrawal. Similar results were obtained if Type 1 and Type 2 diabetic patients were evaluated separately but a delay of the AER reduction was observed in the latter group. In all patients the mean AER was reduced to 60-65% of the initial values. A greater effect of sulodexide on albuminuria was observed in patients with AER above 200 micrograms/min than in those with microalbuminuria (a reduction to 47 vs 65% of the initial output). Sulodexide did not significantly reduce albuminuria in 28% of diabetic patients ('non-responders'). In conclusion, glycosaminoglycan sulodexide may reduce AER in patients with micro or macroalbuminuria and it could slow down development of diabetic nephropathy. PMID- 9347244 TI - Acarbose in ambulatory treatment of non-insulin-dependent diabetes mellitus associated to imminent sulfonylurea failure: a randomised-multicentric trial in primary health-care. Diabetes and Acarbose Research Group. AB - To assess the efficacy and safety of acarbose as an adjunct to high sulfonylurea (SU) doses in patients with imminent SU failure, a randomised, multicentric, 6 month double-blind, parallel and placebo-controlled trial was performed in primary healthcare. Entry criteria were: NIDDM patients in concomitant dietary follow-up, age > 40 year-old, more than 3 years of diagnosed diabetes, baseline HbAlc levels between 8-12% (N: 4-6%), stable body mass index < 35 kg m-2 and glibenclamide daily dose > 10 mg. After 1 month placebo run-in period all patients were randomly allocated into two groups of treatment (acarbose 100 mg t.i.d. vs placebo). HbAlc levels, the main efficacy variable, lipid profile, fasting and postprandial blood glucose levels were performed and adverse events were also recorded. A total number of 65 patients were randomised, 36 in acarbose and 29 in a placebo group. No statistical differences were found on age (60.2/61.7 year-old), BMI (28.7/27.4 kg m-2), glibenclamide dose (14.5/14.0 mg/day) and baseline HbAlc (9.0/8.8%). Acarbose-treated patients significantly reduced HbAlc levels (9.0/7.9 vs 8.8/8.5%; P < 0.01), based upon a marked decrease, but statistically not significant, in mean postprandial plasma glucose levels (11.9/9.6 vs 12.4/11.1 mmol l-1). No significant differences between fasting plasma glucose and lipid profile were detected. A total of 31 patients (47.7%) reported adverse events, 20 (55.5%) and 11 (37.9%) in acarbose and placebo treatment group respectively. Relationship with drug was estimated as possible or probable in 16 (44.4%) of acarbose-treated patients. None of them were excluded from study participation due to insulin requirement. Only seven patients (10.7%), six with acarbose (16.6%) and one with placebo (3.8%), withdrew the study because of the adverse events. Thus, acarbose seems to be a useful option in order to improve HbAlc levels in non-insulin-dependent diabetes mellitus with imminent sulfonylurea failure. PMID- 9347245 TI - Good metabolic and safety profile of troglitazone alone and following alcohol in NIDDM subjects. AB - Drinking alcohol is associated with a recognised risk of hypoglycaemia. This double-blind, placebo-controlled study was designed to determine whether alcohol taken with the evening meal alters the gluco-regulatory response to troglitazone, (TR), an insulin action enhancer, in non-insulin-dependent diabetes mellitus (NIDDM) subjects to increase the risk of hypoglycaemia. In vitro studies conducted prior to the clinical study presented here showed no evidence of a pharmacokinetic interaction between the two drugs. A total of 23, diet-treated, NIDDM subjects received either TR, 200 mg once daily (n = 11) or placebo (PL) (n = 12) for 45 days. On days 42 and 45 subjects were given, on separate days, an alcohol challenge (AC), 0.6 mg/kg ethanol in orange juice and a control challenge, CC, orange juice alone, with the evening meal. Serum glucose, insulin, proinsulin-like molecules, C-peptide and lipids were measured during the study, for the 4 h after each challenge and the following morning (fasting). The over night urine cortisol/creatinine ratio (an index of hypoglycaemia) was also determined. For the TR treated group, fasting serum glucose the next morning (adjusted geometric mean: 6.8 mmol/l for AC) and weighted mean were not statistically significantly different following AC compared to CC. Mean trough glucose for TR after the evening meal was 5.7 mmol/l following both the AC and CC. Analysis of the other parameters showed no symptomatic or pharmacodynamic evidence of an acute interaction between TR and alcohol. It can be concluded that occasional drinking of alcohol, with a meal, by TR-treated NIDDM patients is unlikely to be associated with an increased risk of hypoglycaemia. PMID- 9347247 TI - Comparison of the prevalence in two diabetes surveys in Pu-Li, Taiwan, 1987-1988 and 1991-1992. AB - The objective of this study was to investigate the prevalence of non-insulin dependent diabetes mellitus (NIDDM) in Pu-Li, Taiwan from 1991-1992, and to compare the results with a similar study conducted in 1987-1988. We also wished to compare different approaches in asking about patient history and to determine how this effects data authenticity. Both were community-based cross-sectional studies with stratified cluster sampling of residents age > or = 30. Blood samples were taken for screening and 75 g oral glucose tolerance tests were performed for diagnosis. The total number of eligible subjects in the second study was 2719 (1424 men, 1295 women). Complete data and samples were collected for 1118 (536 men, 582 women). The response rate was 41.1% (37.6% for men, 44.9% for women). The crude prevalence was 10.3% (5.6% known, 4.7% new). Using standard world population (Segi), the age-adjusted prevalence rate was 8.3% (4.0% known, 4.3% new). The 1991-1992 study had a response rate (crude 41.1%, adjusted 51.3%) which was slightly lower than the 1987-1988 study (crude 44.8%, adjusted 55.9%). The age-adjusted prevalence rates for new NIDDM were similar (4.4 vs. 4.3%) while the age-adjusted prevalence of known NIDDM in the second survey (4.0%) was lower than the first survey (6.9%), which apparently was overestimated due to the simplicity of questions regarding history. In conclusion, prevalence of new DM in this area appears to be stable, and when doing a survey regarding previous DM, it is better to include treatment history rather than depending on self-reporting of NIDDM alone. PMID- 9347246 TI - Alterations in serum levels of 1 alpha,25(OH)2 D3 and osteocalcin in patients with early diabetic nephropathy. AB - Serum levels of markers for bone remodeling and diabetic metabolic markers were measured in subjects with non-insulin-dependent diabetes mellitus (NIDDM) to investigate the relationship between early diabetic nephropathy and calcium/bone metabolism. 1 alpha,25(OH)2 D3 (Vit D), osteocalcin (OC), intact parathyroid hormone (PTH) and urine albumin excretion (UAE) were measured in all subjects. Serum levels of Vit D and OC were significantly decreased in diabetic subjects compared to age-matched, non-diabetic controls. In diabetic patients, a significant positive correlation was observed between intact PTH and OC. No significant correlation was found between levels of Vit D and OC. In early diabetic nephropathy without increased serum creatinine, Vit D decreased and OC increased with increasing UAE. Levels of hemoglobin Alc (HbAlc) and fructosamine (FRA) were not correlated with levels of Vit D or OC. Levels of Vit D were decreased and levels of OC were increased in diabetic subjects with proliferative retinopathy or with micro- or macro-albuminuria. CONCLUSIONS: Results of the present study indicate that changes in bone remodeling markers such as Vit D and OC levels are present in the early stages of diabetic nephropathy, and that circulating intact PTH is important in restoring the reduced OC levels in diabetic patients, probably as a reflection of bone remodeling. PMID- 9347248 TI - The effect of pulsatile gonadotropin-releasing hormone and estradiol administration on luteinizing hormone and follicle-stimulating hormone concentrations in pituitary stalk-sectioned ovariectomized pony mares. AB - Hourly pulses of gonadotropin-releasing hormone (GnRH) or bi-daily injections of estradiol (E2) can increase luteinizing hormone (LH) secretion in ovariectomized, anestrous pony mares. However, the site (pituitary versus hypothalamus) of positive feedback of estradiol on gonadotropin secretion has not been described in mares. Thus, one of our objectives involved investigating the feedback of estradiol on the pituitary. The second objective consisted of determining if hourly pulses of GnRH could re-establish physiological LH and FSH concentrations after pituitary stalk-section (PSS), and the third objective was to describe the declining time trends of LH and FSH secretion after PSS. During summer months, ovariectomized pony mares were divided into three groups: Group 1 (control, n = 2), Group 2 (pulsatile GnRH (25 micrograms/hr), n = 3), and Group 3 (estradiol (5 mg/12 hr), n = 3). All mares were stalk-sectioned and treatment begun immediately after stalk-section. Blood samples were collected every 30 min for 8 h on the day before surgery (D0) and 5 d post surgery (D5) to facilitate the comparison of gonadotropin levels before and after pituitary stalk-section. Additionally, jugular blood samples were collected every 12 hr beginning the evening of surgery, allowing for evaluation of the gonadotropin secretory time trends over the 10 d of treatment. On Day 10, animals were euthanized to confirm pituitary stalk-section and to submit tissue for messenger RNA analysis (parallel study). Plasma samples were assayed for LH and FSH by RIA. Mean LH secretion decreased from Day 0 to Day 5 in Groups 1 and 3, whereas LH secretion tended (P < 0.08) to decrease in Group 2 mares. On Day 5, LH was higher (P < 0.01) in Group 2 (17.26 +/- 3.68 ng/ml: LSMEANS = SEM), than either Group 1 (2.65 +/- 4.64 ng/ml) or Group 3 (4.28 +/- 3.68 ng/ml). Group 1 did not differ from Group 3 on Day 5 (P < 0.40). Similarly, mean FSH levels decreased in all groups after surgery, yet Group 2 mares had significantly (P < 0.001) higher FSH concentrations (17.66 +/- 1.53 ng/ml) than Group 1 or Group 3 (8.34 +/- 1.84 and 7.69 +/- 1.63 ng/ml, respectively). Regression analysis of bi-daily LH and FSH levels indicated that the time trends were not parallel. These findings indicate: 1) Pituitary stalk section lowered LH and FSH to undetectable levels within 5 d after surgery. 2) pulsatile administration of GnRH (25 micrograms/hr) maintained LH and FSH secretion, although concentrations tended to be lower than on Day 0, and 3) E2 did not stimulate LH or FSH secretion. PMID- 9347249 TI - Ontogeny of epidermal growth factor (EGF), EGF receptor (EGFR) and basic fibroblast growth factor (bFGF) mRNA levels in pancreas, liver, kidney, and skeletal muscle of pig. AB - Epidermal growth factor (EGF), EGF receptor (EGFR), and basic fibroblast growth factor (bFGF) messenger RNA (mRNA) levels were examined by Northern blot analysis in four tissues (pancreas, liver, kidney, and skeletal muscle) of pig from fetal 90 d to postnatal 180 d of age. The present study shows for the first time that EGF mRNA increased with advancing age in the kidney and skeletal muscle of pig. A high level of EGF mRNA was observed in the kidney compared with the liver and skeletal muscle. In the pancreas, high levels of EGF mRNA were found in fetuses and newborns and were low in older pigs. Pancreatic EGFR mRNA level parallelled its EGF mRNA, whereas in the kidney and skeletal muscle, patterns of EGFR mRNA were reversed to their EGF mRNA levels. In the liver, EGFR mRNA was abundant but EGF mRNA was undetected. In the pancreas and skeletal muscle, the highest levels of bFGF mRNA were found in fetuses of 90 d of age and then decreased with advancing age. In the liver and kidney, there were no major changes in bFGF mRNA levels during the examined developmental periods. These results show that EGF, EGFR, and bFGF mRNA levels are developmentally and tissue specifically regulated in pig. In the pancreas, mRNA levels of EGF, EGFR and bFGF were high in fetal and neonatal life and low thereafter. In the kidney and skeletal muscle, EGF mRNA increased with advancing age. EGF may play a role in muscle growth and maintenance in growing pigs during the later stage of development. PMID- 9347250 TI - cDNA cloning and tissue-specific gene expression of ovine leptin, NPY-Y1 receptor, and NPY-Y2 receptor. AB - The physiological regulation of food intake is a critical factor in both the rate at which an animal grows and its reproductive activity. Recently, progress has been made in elucidating a complex system in which insulin, leptin, and neuropeptide Y function to monitor an animal's energy balance and regulate feed intake and fertility. RNA was extracted from ovine hypothalamic, anterior pituitary, pancreas, and adipose tissue. Using the reverse transcription polymerase chain reaction. cDNAs were cloned and sequenced for leptin (350 base pairs [bp], GenBank accession number U62123 and 441 bp, GenBank accession number U84247), NPY-Y1 receptor (350 bp, GenBank accession no. U62122) and NPY-Y2 receptor (440 bp, GenBank accession no. U83458). Probes generated from these clones were used to detect mRNA expression within tissues thought to be involved in the coregulation of feed intake and reproduction. Leptin was found to be expressed in sheep adipose tissue. The ovine NPY-Y1 receptor mRNA was detected within the arcuate nucleus and paraventricular nucleus of the hypothalamus, the dentate gyrus of the hippocampus and in pancreatic, anterior pituitary, and adipose tissues. Expression of ovine NPY-Y2 receptor mRNA was detected in the hippocampus and within pancreatic tissue. These observations provide evidence of potential mechanisms that exist for mediating communication between peripheral and central tissues within the insulin-leptin-NPY pathway. PMID- 9347252 TI - Serum concentrations of insulin-like growth factors and placental lactogen during gestation in cattle. II. Maternal profiles. AB - This study was designed to examine the effects of fetal growth potential on maternal hormones and lipid metabolism. Sixty beef heifers were inseminated with semen from sires with high (H) or low (L) expected progeny differences for birth weight. Maternal serum was collected at 21-d intervals from Day 85 to Day 274 of gestation. Serum concentrations of insulin-like growth factor (IGF)-I, IGF-II, placental lactogen (PL), and nonesterified fatty acids (NEFA) were determined and correlated to fetal and maternal characteristics. Maternal serum IGF-I declined throughout pregnancy, whereas IGF-II was relatively constant and PL tended to increase. Maternal serum NEFA was low and invariant through Day 211 of gestation when it rose 3.5 times to peak levels at Day 253 and declined at Day 274. PL was positively correlated to NEFA (r = 0.37, P < 0.01), IGF-I was negatively correlated (P < 0.01) to NEFA and PL (r = -0.59, and -0.35, respectively), and IGF-II was negatively correlated to NEFA (r = -0.35, P < 0.01). Dams pregnant with H fetuses had lower (P = 0.02) serum IGF-I and tended to have higher (P = 0.09) serum PL concentrations than dams carrying L fetuses. Additionally, dams pregnant with L fetuses had higher (P < 0.03) serum IGF-II concentrations than dams with H fetuses (175.6 vs. 145.0 ng/ml) during the third trimester. Fetal sex had no effect on any maternal serum parameter. Fetal weight and instantaneous growth rate (IGR) were positively correlated to maternal NEFA and PL and negatively correlated to maternal IGF-I and IGF-II. Independent IGR effects were detected for PL (P < 0.06) and IGF-I (P < 0.0005) concentrations. Maternal hip height was negatively related to serum IGF-I and positively related to serum PL concentrations. Maternal body weight and body condition score were correlated with several serum parameters but were confounded by day of gestation. Correlation analysis of serum concentrations of IGF-I, IGF-II, and PL did not support the hypothesis that PL regulates IGF concentrations. PMID- 9347251 TI - Neural levels of cholecystokinin peptide and mRNA during dietary stimulation of growth and LH secretion in growth-retarded lambs. AB - Chronic feed restriction of prepubertal male lambs adversely affects reproductive development by inhibiting the pulsatile release of luteinizing hormone (LH). Because this effect can be reversed by ad libitum feeding, the associations between diet-induced increases in LH release and concurrent changes in body weight gain, serum glucose. CCK peptide, and CCK mRNA were examined. Neonatally castrated male lambs received a restricted ration to maintain their respective weaning weights beginning at 8 wk of age. At 23 wk of age, lambs were assigned randomly to receive additional feed equivalent to 0%, 50%, or 100% of their previous daily intake. Serum profiles of LH and glucose were determined 2 and 4 wk after onset of the increased intakes. At 27 wk of age, lambs were euthanized and both hypothalamic and cerebral cortical tissues were collected for analysis of CCK peptide and CCK mRNA. With additional intakes, body weight gain increased (P < 0.001) proportional to the graded increases in feed intake. Mean serum LH concentrations, LH peak frequencies, and serum glucose concentrations also increased (P < 0.05) progressively among the 0%, 50%, and 100% dietary intake groups. Neither CCK peptide nor CCK mRNA differed (P > 0.05) among dietary groups suggesting that endogenous CCK in the whole hypothalamus did not change with the feeding-induced increase in LH release. Concentrations of CCK peptide in cerebral cortex were greater (P < 0.05) than hypothalamic concentrations, but there were no differences between hypothalamus and cerebral cortex in the relative abundance of CCK mRNA. In summary, dietary stimulation of growth-retarded male lambs resulted in progressive increases in body weight gain, mean serum LH, serum glucose, and LH peak frequencies. Because hypothalamic levels of CCK peptide and CCK mRNA did not change during feeding-induced secretion of LH, endogenous CCK in the hypothalamus seems unlikely as a chronic mediator of nutrition-sensitive LH release. PMID- 9347253 TI - Effects of an intravenous injection of NPY on leptin and NPY-Y1 receptor mRNA expression in ovine adipose tissue. AB - Neuropeptide Y (NPY) is highly expressed in hypothalami of undernourished and genetically obese animals, and is a potent regulator of food intake and reproduction. Leptin, a protein expressed by adipocytes, has been reported to reduce hypothalamic NPY expression. We recently detected (by ribonuclease protection assay [RPA]) expression of the NPY receptor subtype Y1 (but not Y2) mRNA in adipose tissue. Based on these observations we hypothesized that NPY-Y1 receptors in adipose may represent a peripheral mechanism by which NPY can regulate leptin expression in a direct and rapid manner. To test this hypothesis, adipose samples were biopsied from the tailhead region of 48 +/- 3 kg wether lambs immediately before and 30 min after a single intravenous injection of 50 micrograms porcine NPY ("treated" animals, n = 5), or vehicle ("control" animals, n = 4). Injection of NPY resulted in an increase in expression (P = 0.013; as measured by RPA) of both leptin and NPY-Y1 mRNA. In treated animals, negative correlations were found between response in leptin expression and body weight (r = -0.82, P = 0.092), and between leptin response and initial leptin mRNA levels (r = -0.81, P = 0.097). These data provide evidence of a peripheral mechanism by which NPY may regulate adipocyte expression of both leptin and NPY-Y1 receptor mRNA. PMID- 9347254 TI - Regulation of growth hormone-releasing hormone and somatostatin from perifused, bovine hypothalamic slices. I. Alpha 2-adrenergic receptor regulation. AB - An in vitro perifusion system was developed for bovine hypothalamic tissue to examine the role of alpha 2-adrenergic receptors in the regulation of growth hormone-releasing hormone (GHRH) and somatostatin (SRIF) release. Up to three sagittal slices (600 microns) of hypothalamus, immediately parallel to the midline, were cut in an oxygenated balanced salt solution at 4 degrees C, placed in 5 cc syringes, and perifused at 37 degrees C with oxygenated minimum essential medium-alpha at a flow rate of 0.15 ml/min. Three experiments were conducted, and medium effluent was collected every 20 min before (two samples), during (one or three samples), and after (six samples) treatment. Areas under GHRH and SRIF response curves (AUC), adjusted by covariance for pretreatment values, were calculated from samples collected during the treatment/post-treatment period. Location from which slices were cut, relative to the sagittal midline, had no effect on basal release of GHRH and SRIF, but variation in basal release of GHRH and SRIF differed among animals. Medium containing 60 mM KCI increased AUC for GHRH 39% and 161% for SRIF when compared with perifusion of medium alone, thereby verifying that tissue remained viable for at least 14 hr. Activation of alpha 2 adrenergic receptor with 10(-6) and 10(-4) M clonidine increased AUC for GHRH from 54.8 (control) to 79.1 and 108.7 +/- 2.5 ng.ml-1 min for 10(-6) M and 10(-4) M clonidine, respectively. Guanabenz, another alpha 2-adrenergic receptor agonist, at 10(-8), 10(-6), and 10(-4) M also increased GHRH release from 45.5 (control) to 52.8, 66.2, and 86.7 +/- 1.6 ng.ml-1 min, respectively. Clonidine and guanabenz did not affect release of SRIF. An alpha 2-adrenergic receptor antagonist, idazoxan, blocked clonidine-induced release of GHRH without affecting release of SRIF. We concluded that alpha 2-adrenergic receptor stimulation of in vivo growth hormone secretion in cattle is mediated via an increase in release of GHRH and not a change in release of SRIF. PMID- 9347255 TI - Regulation of growth hormone-releasing hormone and somatostatin from perifused, bovine hypothalamic slices. II. Dopamine receptor regulation. AB - An in vitro perifusion system for bovine hypothalamic tissue was utilized to examine the role of D1 and D2 dopamine receptors in the regulation of somatostatin (SRIF) and growth hormone-releasing hormone (GHRH) release. Up to three sagittal slices (600 microns) of bovine hypothalamus, immediately parallel to the midline, were cut in an oxygenated balanced salt solution at 4 degrees C, placed in 5 cc syringes, and perifused at 37 degrees C with oxygenated minimum essential medium-alpha at a flow rate of 0.15 ml/min. Five experiments were conducted, and medium effluent was collected every 20 min before (two samples), during (one or three samples), and after (six samples) treatment. Areas under SRIF and GHRH response curves (AUC), adjusted by covariance for pretreatment values, were calculated from samples collected during the treatment/post treatment period. Activation of D1 receptor with 10(-8) M and 10(-6) M SKF 38393 increased AUC for SRIF from 5.6 (control) to 420 and 500 +/- 57.8 ng.ml-1 min, but 10(-10) M SKF 38393 was ineffective. Relative to controls, release of GHRH was decreased 50% in the 10(-6) M SKF 38393 group. Blockade of D1 receptors with SCH 23390 had no effect on basal release of either SRIF or GHRH, but prevented SKF 38393-induced release of SRIF and SKF 38393-induced suppression of GHRH. In contrast, quinelorane, a D2 receptor agonist, and haloperidol, which blocks D2 receptors, did not affect release of SRIF or GHRH. We concluded that activation of D1 dopamine receptors, but not D2 dopamine receptors, stimulates release of SRIF and inhibits release of GHRH from the bovine hypothalamus. PMID- 9347256 TI - Regulation of growth hormone-releasing hormone and somatostatin from perifused, bovine hypothalamic slices. III. Reciprocal feedback between growth hormone releasing hormone and somatostatin. AB - An in vitro perifusion system for bovine hypothalamic tissue was used to determine if growth hormone-releasing hormone (GHRH) and somatostatin (SRIF) modulate each other's release, and whether SRIF mediates D1-agonist-induced suppression of GHRH in cattle. Up to three sagittal slices (600 microns) of bovine hypothalamus, immediately parallel++ to the midline, were cut in an oxygenated balanced salt solution at 4 degrees C, placed in 5 cc syringe barrels, and perifused at 37 degrees C with oxygenated minimum essential medium-alpha at a flow rate of 0.15 ml/min. Three experiments were conducted, and medium effluent was collected every 20 min before (two samples), during (one or three samples), and after (six samples) treatment. Areas under GHRH and SRIF response curves (AUC), adjusted by covariance for pretreatment values, were calculated from samples collected during the treatment/post-treatment period. Perifusion of SRIF at 10(-6) M and 10(-4) M decreased AUC for GHRH from 86.3 (control) to 65.4 and 59.5 +/- 6.3 ng.ml-1 min, but 10(-8) M SRIF was ineffective. Relative to controls, 10(-8).10(-6), and 10(-4) M GHRH increased release of SRIF 190, 675, and 1,135%, respectively. Activation of D1 receptors with 10(-6) M SKF 38393 increased AUC for SRIF from 12.5 ng.ml-1 min (control) to 484.9 ng.ml-1 min and decreased AUC for GHRH from 36.4 ng.ml-1 min (control) to 18.2 ng.ml-1 min. Blockade of SRIF action with a SRIF antagonist, cyclo-[7-aminoheptanoyl-phe-D-trp lys-thr(bzl)], increased release of GHRH 1.9-fold. In addition, the SRIF antagonist blocked SKF 38393-induced suppression of GHRH. We concluded that GHRH and SRIF interact within the bovine hypothalamus/pituitary stalk to modulate the release of the other. Moreover, SRIF mediates the inhibitory effects of activation of D1 receptors on release of GHRH in cattle. PMID- 9347257 TI - PRIME II (Second Prospective Randomized Study of Ibupamine on Mortality and Efficacy): another disappointment in heart failure therapy. PMID- 9347258 TI - Smoking and outcome after PTCA. PMID- 9347259 TI - Distribution of knowledge. PMID- 9347260 TI - Validation for coronary stenting: a permanent implant for interventional cardiology. PMID- 9347262 TI - A 'phone call to heaven. Is the cellular 'phone dangerous for its user with a pacemaker? PMID- 9347261 TI - The ultimate interventional cardiologist--a computer. PMID- 9347263 TI - Do ACE inhibitors modulate atherosclerosis? AB - The ACE/angiotensin II/bradykinin system is inextricably linked to some of the processes that contribute to the generation of atherosclerosis at genetic, molecular, biochemical and pharmacological levels. There is a large body of laboratory-derived experimental data that suggests that inhibition of ACE activity has antiproliferative, anti-inflammatory and vasodilatory effects that can modulate this atherosclerotic process from the earliest form of endothelial dysfunction, to delay of lesion formation in primary atherosclerosis or in myointimal proliferation after PTCA. The clinical evidence for these potential benefits is so far sparse. There are several possible explanations for these discrepancies. Firstly, the role of the ACE/bradykinin/angiotensin II system in the local vascular response to either the primary process of atherosclerosis, or to the injury induced by balloon angioplasty is likely to vary between species and models. Secondly, there is a tendency to ensure the presence of ACE inhibitor in high concentration before or during the vascular insult in animal models, whereas this has not been the case in the clinical studies of post-PTCA restenosis. Whilst the animal studies therefore offer potentially valuable insights into the mechanics of local vascular response, the ability of ACE inhibitors to interfere with such mechanisms now needs to be tested in clinical trials that are each aimed at precisely answering specific questions. The experimental data so far lend considerable support to the fact that drugs acting solely by interference with the angiotensin II-receptor complex are at a theoretical disadvantage, when compared with ACE inhibitors, since the former would be expected to have little effect on bradykinin-mediated activities. To the established benefits of ACE inhibitors in left ventricular dysfunction, and the interesting possibility that there may be an anti-ischaemic action in these circumstances, we may add the promise of the TREND study. In the coming years, there is an urgent requirement for intensive investigation into the ability of ACE inhibitors to modulate the various stages of the atherosclerotic spectrum. For now though, the jury remains out. PMID- 9347264 TI - Recommendations on stent manufacture, implantation and utilization. Study Group of the Working Group on Coronary Circulation. PMID- 9347265 TI - Doppler evaluation of left ventricular filling in congestive heart failure. PMID- 9347266 TI - Innervation and effects of vasoactive substances in the coronary circulation. PMID- 9347267 TI - EUROASPIRE. A European Society of Cardiology survey of secondary prevention of coronary heart disease: principal results. EUROASPIRE Study Group. European Action on Secondary Prevention through Intervention to Reduce Events. AB - BACKGROUND: The three major European scientific societies in cardiovascular medicine--the European Society of Cardiology (ESC), the European Atherosclerosis Society and the European Society of Hypertension--published in October 1994 joint recommendations on prevention of coronary heart disease in clinical practice. Patients with established coronary heart disease, or other major atherosclerotic disease, were deemed to be the top priority for prevention. A European survey (EUROASPIRE) was therefore conducted under the auspices of the ESC to describe current clinical practice in relation to secondary prevention of coronary heart disease. AIMS: The aims of EUROASPIRE were (i) to determine whether the major risk factors for coronary heart disease are recorded in patients medical records; (ii) to measure the modifiable risk factors and describe their current management following hospitalization, and (ii) to determine whether first degree blood relatives have been screened. METHODS: The survey was conducted in selected geographical areas and hospitals in nine European countries. Consecutive patients (< or = 70 years) were identified retrospectively with the following diagnoses: coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, acute myocardial infarction and acute myocardial ischaemia without infarction. Data collection was based on a retrospective review of hospital medical records and a prospective interview and examination of the patients. RESULTS: 4863 medical records were reviewed of whom 25% were women, and 3569 patients were interviewed (adjusted response rate 85%) with an average age of 61 years. Nineteen percent of patients smoked cigarettes, 25% were overweight (BMI > or = 30 kg.m-2), 53% had raised blood pressure (systolic BP > or = 140 and/or diastolic BP > or = 90 mmHg), 44% had raised total plasma cholesterol (total cholesterol > or = 5.5 mmol.l-1) and 18% were diabetic. Reported medication at interview was: antiplatelet drugs 81%, beta-blockers, 54% (58% in post-infarction patients). ACE inhibitors 30% (38% in post infarction patients) and lipid lowering drugs 32%. Of the patients receiving blood pressure lowering drugs (not always prescribed for the treatment of hypertension) 50% had a systolic BP > 140 mmHg and 21% > 160 mmHg, and of those receiving lipid lowering drugs, 49% had plasma total cholesterol > 5.5 mmol.l-1 and 13% > 6.5 mmol.l-1. Thirty-seven percent of patients had a family history of premature coronary heart disease in a first-degree blood relative, but only 21% of patients reported being advised to have their relatives screened for coronary risk factors. CONCLUSIONS: This European survey has demonstrated a high prevalence of modifiable risk factors in coronary heart disease patients. There is considerable potential for cardiologists and physicians to further reduce coronary heart disease morbidity and mortality and improve patients chances of survival. PMID- 9347268 TI - Sex-related differences in eligibility for reperfusion therapy and in-hospital outcome after acute myocardial infarction. AB - AIMS: To determine the effect of sex on reperfusion therapy and early mortality after acute myocardial infarction. METHODS: We analysed the characteristics, the reperfusion interventions, and in-hospital mortality in 400 consecutive patients (320 men and 80 women) admitted during the first 6 h of acute myocardial infarction and treated by primary angioplasty, or intravenous thrombolysis with rescue angioplasty. RESULTS: The differences between men and women were age (57 vs 67 years, P = 0.001), systemic hypertension (33 vs 50%, P = 0.02), cigarette smoking (79 vs 30%, P = 0.0001) and contraindications to thrombolysis (28.5 vs 42.5%, P = 0.02). Successful reperfusion of the infarct-related artery was achieved in 84% of patients of both sexes. In-hospital mortality was 7.2% in men and 18.7% in women (P = 0.001). Multivariate analysis was performed by linear logistic regression in order to compare several embedded models, using repeated maximum likelihood ratio tests. The best model involved the variables of cardiogenic shock and age. Addition of the variable 'sex' did not improve the predictive power of this model (P > 0.5). CONCLUSION: During acute myocardial infarction, similar successful early reperfusion rates can be achieved in men and women, despite the lower eligibility of women for thrombolytic therapy. Although in-hospital mortality was higher in women than men, the best predictive model of mortality was the combination of age and cardiogenic shock. Therefore, sex does not appear to be an independent predictor of mortality. PMID- 9347269 TI - Effects of atrial pacing stress test on ultrasonic integrated backscatter cyclic variations in normal subjects and in patients with coronary artery disease. AB - AIMS: To evaluate the effects of acute, atrial pacing-induced, reversible myocardial ischaemia on myocardial thickening and integrated backscatter cyclic variations in patients with or without coronary artery disease. METHODS AND RESULTS: Thirty-six patients with suspected coronary artery disease underwent transoesophageal echocardiography with simultaneous atrial pacing, and coronary angiography. In myocardial segments not related to a significantly narrowed coronary artery, both from patients with and without coronary artery disease, thickening and integrated backscatter cyclic variations were not reduced at peak pacing. In segments related to a significantly narrowed coronary artery, thickening decreased at peak pacing, was still reduced at pacing interruption and recovered at 2 min, while backscatter cyclic variations, blunted at peak pacing, immediately recovered after pacing interruption. CONCLUSION: During stress induced myocardial ischaemia, backscatter cyclic variations are blunted and thickening reduced. Returning to baseline, pre-atrial pacing values occur more rapidly in backscatter cyclic variations than when thickening takes place. Evaluation of stress-induced alterations in backscatter cyclic variations may aid in the identification of ischaemia-induced regional left ventricular functional impairment and, hence, in coronary artery disease diagnosis. PMID- 9347270 TI - Prognostic value of left ventricular volume response during dobutamine stress echocardiography. AB - AIMS: An abnormal left ventricular volume response during dobutamine echocardiography identified patients with severe coronary artery disease. The aim of the study was to assess the prognostic value of left ventricular volume changes during dobutamine stress echocardiography in 136 patients. METHODS AND RESULTS: Endpoints were defined as spontaneous cardiac events at follow-up. Left ventricular end-diastolic and end-systolic volume changes (abnormal response: < 10% and < 20% decrease, respectively) were compared with other clinical and stress test variables. During 18 +/- 7 months of follow-up, 31 cardiac events occurred: 12 hard events (cardiac death [n = 6], myocardial infarction [n = 6]) and 19 soft events (unstable angina [n = 16], congestive heart failure [n = 3]). End-diastolic volume response (P = 0.006), diabetes (P = 0.008), inducible wall motion abnormalities (P = 0.024), end-systolic volume response (P = 0.039) and inducible angina (P = 0.038) were related to a greater likelihood of cardiac events. The Cox regression analysis revealed end-diastolic volume response (odds ratio: 3.0; CI 1.44-6.32) and diabetes (odds ratio: 2.7; CI 1.28-5.69) to be independent predictors of spontaneous cardiac events. Diabetes (odds ratio: 4.0; CI 1.26-12.80) and < 40% baseline ejection fraction (odds ratio: 2.21; CI1.14 4.29) were independent predictors of hard events. CONCLUSION: An abnormal end diastolic volume response during dobutamine stress echocardiography identifies patients with an unfavourable outcome; they should be considered for more accurate prognostic stratification. PMID- 9347271 TI - Severe mitral regurgitation complicating acute myocardial infarction. Clinical and angiographic differences between patients with and without papillary muscle rupture. AB - AIMS: To assess the differential clinical and angiographic characteristics of patients with severe mitral regurgitation related (n = 31) or unrelated (n = 16) to papillary muscle rupture complicating acute myocardial infarction. METHODS AND RESULTS: The clinical and angiographic features of patients with myocardial infarction and severe mitral regurgitation were evaluated. Patients with papillary muscle rupture were older (67 vs 60 years, P < 0.005) and had a lower rate of diabetes (7% vs 38%, P < 0.005) and of previous angina or infarction (24% vs 50%, P < 0.05). Frequency of inferior infarction was high and comparable in both groups (papillary muscle rupture, 72% vs non-papillary muscle rupture, 88%, ns) whereas in-hospital rate of angina/infarct extension prior to mitral regurgitation, also high, tended to be higher in patients without than in those with papillary muscle rupture (67% vs 39%, ns). Incidence of multivessel disease tended to be higher in patients without papillary muscle rupture (87% vs 56%, P < 0.06) and they had a lower ejection fraction (46 +/- 15 vs 61 +/- 14%, P < 0.03), whereas the culprit artery was mainly the right or the circumflex coronary artery in both groups (papillary muscle rupture, 100% vs non papillary muscle rupture, 93%, ns). Valve replacement was performed earlier in patients with papillary muscle rupture (1 (1; 14) vs 25 (5; 45) days, median, P < 0.002) but was associated with a similar mortality (papillary muscle rupture 11/24, 46% vs non papillary muscle rupture, 7/15, 47%, ns). The main cause of death was cardiogenic shock in patients without papillary muscle rupture (5/7, 71%), and respiratory insufficiency--sepsis in those with papillary muscle rupture (7/11, 64%). CONCLUSIONS: Severe mitral regurgitation in myocardial infarction with or without papillary muscle rupture is mostly related to inferior infarction and often follows reinfarction, particularly in non-papillary muscle rupture cases. The main contributors to surgical mortality appear to be respiratory insufficiency in patients with papillary muscle rupture and cardiogenic shock, facilitated by a lower ejection fraction, a higher frequency of diabetes and more extensive coronary disease, in patients without papillary muscle rupture. PMID- 9347272 TI - A prognostic computer model to predict individual outcome in interventional cardiology. The INTERVENT Project. AB - It is not yet possible to predict an individual's outcome from percutaneous transluminal coronary angioplasty or alternative/adjunctive coronary interventional techniques. The purpose of the INTERVENT project is to redefine complications associated with coronary interventions, to set up a prognostic computer model to predict individual outcome and to compare the results to those of conventional statistical techniques. 2500 data items were analysed in 455 consecutive patients (mean age: 61.1 +/- 8.3 years; range 33-84 years; 80.4% male, 16.7% unstable angina, 5.1%/10.1% acute/subacute myocardial infarction) undergoing coronary interventions at three university centres. In-lab/out-of-lab complication rates were 0.4%/0.9% (death), 1.8%/0.2% (abrupt vessel closure with myocardial infarction) and 5.5%/4.0% (haemodynamic complications). Computer algorithms derived by applying techniques from artificial intelligence were able (1) to reduce the set of possible relevant risk factors from 2500 to about 40, (2) to predict individual risk with an accuracy of > 95% and (3) to explain the structural relationship between outcome and risk factors. Patient data from two centres were used to construct and test the algorithm. Data from a third centre were used to evaluate the algorithm. The most important predictors-were acute myocardial infarction, heart failure (NYHA class > II), unstable angina, complex lesions, high low density lipoprotein cholesterol and duration of coronary heart disease. Neither age nor gender impaired the percutaneous transluminal coronary angioplasty results in acute ischaemic syndromes; however, for stable angina, procedural risk increased with age. There was little risk from primary percutaneous transluminal coronary angioplasty in acute myocardial infarction in patients with NYHA heart failure classes I-II; however, the risk was high for patients in NYHA classes > II, either with or without additional thrombolysis. Alternative/adjunctive intervention techniques were no predictors for in-lab-, but were predictors for post-procedural complications. PMID- 9347273 TI - Haemodynamic, neuroendocrine and metabolic correlates of circulating cytokine concentrations in congestive heart failure. AB - OBJECTIVES: Increased activity of pro-inflammatory cytokines in the circulation has been observed in many, though not all, patients with congestive heart failure. To identify the predictors of cytokine activation in congestive heart failure, we assessed the relationship of peripheral and hepatic venous cytokines to central haemodynamics, neuroendocrine status and intermediary metabolism in patients with moderate or severe congestive heart failure. PATIENTS AND METHODS: Concentrations of tumour necrosis factor-alpha, soluble tumour necrosis factor receptor II and interleukin 6 were measured from peripheral and hepatic venous plasma in 58 adult cardiac patients, of whom 44 had congestive heart failure, undergoing heart catheterization, echocardiography and assessment of selected neuroendocrine and metabolic characteristics. RESULTS: Peripheral venous soluble tumour necrosis factor-receptor II was directly related to NYHA class (rs = 0.46, P < 0.001) and inversely to 6-min walking distance (rs = -0.46, P < 0.001). Peripheral venous tumour necrosis factor-alpha was related to 6-min walking distance (rs = -0.37, P < 0.01), but like soluble tumour necrosis factor-receptor II, was unrelated to other haemodynamic and neuroendocrine measurements. Peripheral venous interleukin 6 correlated with NYHA class (rs = 0.66, P < 0.001) and 6-min walking distance (rs = -0.52, P < 0.001). In addition, interleukin 6 was related to right atrial pressure (rs = 0.55, P < 0.001), pulmonary artery wedge pressure (rs = 0.50, P < 0.001) and left ventricular ejection fraction (rs = -0.39, P < 0.01); in multivariate analysis, only right atrial pressure was an independent predictor of interleukin 6 concentration (P < 0.001). Comparisons between patients with and without congestive heart failure showed significantly higher hepatic venous tumour necrosis factor-alpha, soluble tumour necrosis factor-receptor II and interleukin 6 in the heart failure group; the differences in peripheral venous cytokines were less consistent. CONCLUSIONS: In cardiac patients, increased plasma tumour necrosis factor-alpha and soluble tumour necrosis factor-receptor II are associated with symptoms of heart failure and poor exercise capacity, while the most important predictor of increased interleukin 6 is elevated systemic venous pressure. Different but still unknown mechanisms may be responsible for the increased release of cytokines in congestive heart failure. PMID- 9347275 TI - Influence of digital and analogue cellular telephones on implanted pacemakers. AB - The aim of this study was to find out whether digital and analogue cellular 'phones affect patients with pacemakers. The study comprised continuous ECG monitoring of 200 pacemaker patients. During the monitoring certain conditions caused by interference created by the telephone were looked for: temporary or prolonged pacemaker inhibition; a shift to asynchronous mode caused by electromagnetic interference; an increase in ventricular pacing in dual chamber pacemakers, up to the programmed upper rate. The Global System for Mobile Communications system interfered with pacing 97 times in 43 patients (21.5%). During tests on Total Access of Communication System telephones, there were 60 cases of pacing interference in 35 patients (17.5%). There were 131 interference episodes during ringing vs 26 during the on/off phase; (P < 0.0001); 106 at maximum sensitivity level vs 51 at the 'base' value; P < 0.0001). Prolonged pacing inhibition (> 4 s) was seen at the pacemaker 'base' sensing value in six patients using the Global system but in only one patient using Total Access. CONCLUSION: Cellular 'phones may be dangerous for pacemaker patients. However, they can be used safely if patients do not carry the 'phone close to the pacemaker, which is the only place where high risk interference has been observed. PMID- 9347274 TI - Skeletal muscle function at low work level as a model for daily activities in patients with chronic heart failure. AB - AIM: Metabolic exercise abnormalities have been reported in chronic heart failure patients. This study sought to evaluate whether these abnormalities affected daily activity. METHODS AND RESULTS: In 16 patients with moderate-to-severe chronic heart failure and in eight controls we measured femoral flow (thermodilution) and metabolism (glucose, lactate, free fatty acids, blood gas values) at rest and during a constant load of 20 W, which may mimic a daily activity. At rest, chronic heart failure patients had a leg flow similar to controls, but showed a higher leg oxygen consumption (4.6 +/- 0.6 vs 2.6 +/- 0.4 ml.min-1; P < 0.05), a higher arteriovenous oxygen difference (7.2 +/- 0.5 vs 5.4 +/- 0.7 ml.dl-1; P < 0.05), and a lower femoral vein pH (7.37 +/- 5.03 vs 7.42 +/ 0.01; P = 0.01). At 20 W, chronic heart failure patients had a leg flow similar to controls, but showed increased lactate release (from resting 11.7 +/- 33 to 142 +/- 125 micrograms.min-1 P < 0.0001 vs controls, from resting 5.7 +/- 15.4 to 50 +/- 149 micrograms.min-1 ns), higher arterial concentration of free fatty acids (781 +/- 69 vs 481 +/- 85 mumol.l-1; P < 0.01), lower femoral vein HCO3 (24.1 +/- 2.6 vs 26.3 +/- 1.7 mmol.l-1; P < 0.05) and base excess (-2.3 +/- 2.3 vs -0.24 +/- 1.7 mmol.l-1; P = 0.01). CONCLUSION: In chronic heart failure patients, the important cellular metabolic alterations already present at rest partially affect daily activities, owing to a further decrease in the efficiency of muscle metabolic processes, and may preclude tolerance of heavier activities. Such alterations appear, at least in part, independent of peripheral haemodynamic responses to exercise. PMID- 9347276 TI - Heart rate dependency of premature ventricular contractions. Correlation between electrocardiographic monitoring and exercise-related patterns. AB - AIMS: The identification of the response of ventricular arrhythmias to exercise testing could provide important information in the clinical setting but is difficult to obtain as no definite criteria are available. The aim of this study was to evaluate whether analysis of spontaneous heart rate dependency of premature ventricular contractions evaluated by means of 24 h electrocardiographic monitoring is capable of predicting their behaviour during exercise. METHODS AND RESULTS: One hundred and twenty-six patients (49 +/- 16 years) with frequent (> 30 h) premature ventricular contractions underwent 24 h electrocardiographic monitoring in order to evaluate the relationship between arrhythmias and heart rate, and an exercise test in order to evaluate the behaviour of the arrhythmias. On the basis of the incidence of premature ventricular contractions at different cardiac cycle lengths found at electrocardiographic monitoring, three groups of patients were identified: 34 with a tachycardia-enhanced pattern (the shorter the cycle length the higher the incidence of arrhythmias); 32 with a bradycardia-enhanced pattern (the longer the cycle length the higher the incidence of arrhythmias); and 60 patients with an indifferent pattern (no relationship). During the exercise test, the number of premature ventricular contractions/min at maximal effort in comparison with baseline increased in patients with a tachycardia-enhanced pattern (couplets or runs appeared in 10), decreased in those with a bradycardia-enhanced pattern and did not change in those with an indifferent pattern. The positive predictive accuracy of the tachycardia-enhanced pattern in predicting an increase in premature ventricular contractions > 100% at maximal effort was 78%. CONCLUSION: Identification of spontaneous behaviour between the incidence of premature ventricular contractions and the length of the preceding cardiac cycle may predict the behaviour of arrhythmias during exercise. PMID- 9347277 TI - Value of single oral loading dose of propafenone in converting recent-onset atrial fibrillation. Results of a randomized, double-blind, controlled study. AB - AIMS: To evaluate the efficacy and safety of a single loading oral dose of propafenone in the interruption of recent-onset atrial fibrillation. METHODS: After a complete medical history, physical examination, 12-lead ECG, chest X-ray, and routine biochemical laboratory testing, 55 consecutive patients with recent onset atrial fibrillation were randomized double-blind in the emergency department for the administration of either a single oral dose (450 to 750 mg) of propafenone (29 cases) or a placebo (26 cases). After the 24-h observation period, comprehensive echocardiographic examination was performed. RESULTS: The groups were homogeneous as regards biological, clinical and echocardiographic characteristics. Two hours after treatment, 12 patients (41%) on propafenone but only two (8%) on placebo had converted to sinus rhythm (P = 0.005). This striking difference was maintained 6 h after treatment (65 vs 31%; P = 0.015) but lessened at 12 h (69% vs 42%; P = 0.060) and was insignificant at the end of the 24-h treatment period (79%, vs 73%; P = 0.752). Apart from hypotension, transient in three cases and sustained in one whose later echocardiographic examination demonstrated left systolic ventricular dysfunction, propafenone was well tolerated. CONCLUSION: Although there is no significant difference in the rates of conversion 24 h after treatment, propafenone works faster than placebo in achieving sinus rhythm. This rapid action of oral propafenone can be useful to solve quickly the clinical problems of a high proportion of patients arriving at the emergency department with acute atrial fibrillation. PMID- 9347278 TI - Effect of the addition of an abdominal hot can cardioverter/defibrillator pulse generator on the defibrillation energy requirements in a single-lead endocardial defibrillation system. AB - AIMS: The effects of a cardioverter/defibrillator system with an electrically active generator can, applied without recourse to thoracotomy, have not been investigated in the abdominal position in humans. The purpose of this acute clinical study was to evaluate the defibrillation efficacy of an abdominally positioned hot can electrode in connection with a single lead endocardial defibrillation system. PATIENTS AND METHODS: Thirty consecutive patients undergoing implantation of a cardioverter/defibrillator or pulse generator replacement were enrolled in this study Each patient received an integrated, tripolar single-lead system. This was tested using an asymmetrical biphasic defibrillation waveform with constant energy delivery. Defibrillation energy, peak voltage, peak current and impedance were compared between two electrode configurations: (A) in this configuration the distal right ventricular coil was negative and the proximal coil positive; (B) in this configuration the distal right ventricular coil was negative and the proximal coil and the abdominal hot can (65 ccm), as common anode, were positive. Defibrillation threshold testing started at 15 J with stepwise energy reduction (10 J, 8 J, 5 J and 3 J) until defibrillation was ineffective. RESULTS: Compared to the single-lead configuration, the abdominal hot can configuration revealed at 17.5% reduction in defibrillation energy requirements (8.6 J +/- 4.3 J vs 10.43 J +/- 3.9 J; P = 0.041), a 15.7% reduction in peak voltage (308.6 V +/- 63 V vs 365.3 V +/- 68 V; P = 0.003), and a 21.6% reduction in impedance (41.1 omega +/- 6.3 omega vs 52.4 omega +/- 6.6 omega; P < 0.001). Peak current showed a significant increase during hot can testing of 8.2% (7.2 A +/- 1.8 A vs 7.8 A +/- 2.2 A; P = 0.16). CONCLUSION: An abdominally placed hot can pulse generator lowered defibrillation energy requirements in patients with an endocardial defibrillation lead system. PMID- 9347279 TI - Pregnancy in patients with pulmonary autograft valve replacement. AB - AIM: The purpose of this study is to determine the outcome and complications of pregnancy in women with pulmonary autograft valve replacement for aortic valve disease. METHODS AND RESULTS: The records of all women who had undergone pulmonary autograft valve replacement at the National Heart Hospital (now Royal Brompton Hospital) since 1968 were reviewed. From 1968 to 1993, 27 hospital survivors were female and among eight of them there were 14 pregnancies. All women were in Ability Index 1 at time of pregnancy with normal ventricular function, mild aortic regurgitation (six), mild pulmonary regurgitation (three) and mild pulmonary stenosis (two). None took anticoagulants. There was no maternal death, thromboembolic or haemorrhagic event or evidence of deterioration in valve function during pregnancy. Except for one woman (Ability Index 3) who developed dilated cardiomyopathy without aortic or pulmonary valve disease 6 months after delivery, the women remained in Ability Index 1 after pregnancy. There was no significant progression of aortic regurgitation (mild after seven pregnancies), pulmonary regurgitation (mild after six) or right-sided obstruction (mild after four). Reoperation for right-sided obstruction was carried out in two patients 4 and 7 years after a second pregnancy (9 and 15 years after the pulmonary autograft). CONCLUSION: No valve-related complications occurred during pregnancy and pregnancy appeared to have no effect on the function of the pulmonary valve autograft or the right-sided homograft. The pulmonary autograft is thus an ideal procedure for a young female needing aortic valve replacement. PMID- 9347280 TI - Influence of heart rate, respiration and recipient atrial contraction on pulsed wave transmitral Doppler flow indices in orthotopic heart transplant recipients. AB - AIMS: The study set out to assess the relative contributions of donor heart rate, respiration and recipient atrial contraction on the mean of pulsed wave transmitral Doppler flow indices in orthotopic heart transplant recipients. This would provide information on the theoretical usefulness of pacemaker synchronization of recipient atrial contraction, as well as on the validity of certain strategies used for pulsed wave Dopper analysis of diastolic left ventricular function, which have excluded beats based on recipient atrial contraction timing. METHODS: Thirty two consecutive patients undergoing orthotopic heart transplantation in our centre were prospectively studied. The following Doppler indices were analysed: peak early diastolic velocity (E) and its area under the Doppler curve (TVIE), diastolic velocity after donor atrial contraction (A) and its area under the curve (TVIA), the total area under the curve (TVI), the isovolumic relaxation period (IVR), the diastolic filling period, the normalized peak filling rate and the pressure half time. RESULTS: Only 81 out of 347 recordings (23%) allowed analysis of the recipient P wave and thus recipient atrial contraction timing, heart rate and the respiration phase in 22 patients for a total of 1579 beats. The isovolumic relaxation period, E, pressure half time and TVIA are not influenced by donor heart rate. For the isovolumic relaxation period, E, TVI and TVIE, respiration contributes as much as recipient atrial contraction timing to beat-to-beat variation. Pressure half time, the diastolic filling period and peak filling rate were not affected by respiration. TVI was not affected by recipient atrial contraction timing. CONCLUSION: With respect to analysis of diastolic function, exclusion of beats based on recipient atrial contraction timing is invalid for the isovolumic relaxation period. E, TVI and TVIE, since these are equally influenced by respiration. Since TVI was not affected by recipient atrial contraction timing, pacemaker synchronization of donor and recipient atria is not expected to be useful in patients with left ventricular diastolic dysfunction. PMID- 9347281 TI - Systolic blood pressure and (cardiac) mortality over 15 years after venous coronary bypass surgery. AB - OBJECTIVE: The aim of the present study was to determine the influence of pre operative systolic blood pressure and systolic blood pressure 1 and 5 years after venous coronary bypass surgery on subsequent cardiac and non-cardiac mortality. DESIGN: A prospective 15 years follow-up study. PATIENTS: A series of 446 consecutive coronary bypass surgery patients, operated on between April 1976 and April 1977. According to their systolic blood pressure, patients were divided into five groups. MAIN OUTCOME MEASURES: Systolic blood pressure 5 years after surgery, but not pre-operative systolic blood pressure, was an independent predictor of cardiac mortality. RESULTS: Multivariate Cox proportional hazards analysis revealed that pre-operative systolic blood pressure was not associated with cardiac mortality, while higher systolic blood pressure 1 year after surgery showed a trend towards increased cardiac mortality. Systolic blood pressure 5 years after surgery appeared to be a strong independent predictor of cardiac mortality during the subsequent follow-up period. Patients with a systolic blood pressure of 130-139 mmHg had the lowest risk. Compared to this group, the cardiac mortality risk in patients with a systolic blood pressure 5 years after surgery of 140-149 mmHg, 150-159 mmHg and > or = 160 mmHg, was 2.3 (1.2 to 4.6), 3.4 (1.6 to 7.1) and 3.1 (1.4 to 6.5) times higher. Systolic blood pressure < 130 mmHg 5 years after surgery was also associated with a 2.3 times (1.1 to 4.7) times increased risk for cardiac mortality, compared to patients with a systolic blood pressure of 130-139 mmHg. CONCLUSIONS: These findings underline the importance of systolic blood pressure control in the initial years after coronary bypass surgery. PMID- 9347282 TI - Metallothionein free radical scavenger and cardiovascular morbidity. PMID- 9347283 TI - Paradoxical acute thromboembolism during prostacyclin administration. PMID- 9347284 TI - Folate intake in Europe: recommended, actual and desired intake. AB - OBJECTIVES: To evaluate possible inconsistencies between recommended, actual and desired folate intake in European adult populations. DESIGN: Review of dietary recommendations, of food consumption surveys, and of intervention and observational studies relating folate intake to the risk of neural tube defects and plasma homocysteine levels. RESULTS: In Europe, mean dietary folate intake in adults is 291 micrograms/d (range 197-326) for men and 247 micrograms/d (range 168-320) for women. The recommended intakes vary between 200-300 micrograms/d (men) and 170-300 micrograms/d (women). However, women with a previous pregnancy affected by a neural tube defect (NTD), are recommended to take 4000 micrograms/d of supplemental folic acid when planning a subsequent pregnancy. For those without a history of NTD, the use of 400 micrograms/d of supplemental folic acid is the best option to prevent the occurrence of NTDs. A daily dose of 650 micrograms supplemental folic acid normalises elevated plasma homocysteine levels, which is a risk factor for cardiovascular diseases. A dietary folate intake of at least 350 micrograms/d is desired to prevent an increase in plasma homocysteine levels of the adult population in general. CONCLUSIONS: Mean dietary folate intake in Europe is in line with recommendations, but the desired dietary intake of > 350 micrograms/d is only reached by a small part of studied European populations. It is considered unethical to investigate whether supplements with a dose lower than 400 micrograms/d of folic acid are also protective against NTDs. However, research to establish the lowest effective dose of dietary folate/supplemental folic acid to optimise homocysteine levels and research on the bioavailability of folate is required. This will enable the choice of a strategy to achieve desired folate intakes in the general population. In the meantime, consumption of plant foods like vegetables, fruits, and cereals should be stimulated to reach the desired level of 350 micrograms of dietary folate per day. PMID- 9347287 TI - Body fatness and bioelectrical impedance in non-obese pre-menarcheal girls: comparison to anthropometry and evaluation of predictive equations. AB - OBJECTIVES: To determine in non-obese pre-menarcheal girls if bioelectrical impedance (BIA) is a better predictor of body fatness than triceps skinfold (TSF) or body mass index (BMI) and to cross-validate published equations for determination of fat-free mass (FFM) from BIA in pre-menarcheal girls. DESIGN: Cross-sectional analysis of data from 132 non-obese pre-menarcheal girls. The relationship of percent body fat (%BF), derived from isotopic dilution of H2 18O to TSF, BMI, and %BF by BIA, calculated from measures of height, weight and resistance was examined by correlation analysis. SETTING: Massachusetts Institute of Technology (MIT) Clinical Research Center in Cambridge, MA, USA. SUBJECTS: Pre menarcheal girls aged 8-12 y were recruited from local schools, MIT summer day camp and by word of mouth. RESULTS: TSF accounted for 68% of the explained variance (R2) in the prediction of %BF measured by H2 18O, compared to 38% for BMI and 70% for BIA. Prediction of FFM by comparison of published equations was evaluated in this population. The predictive ability differed by Tanner stage. Kushner's equation (Kushner et al, 1992), based solely on height2/resistance was the only equation that provided estimates that did not differ significantly from measured values among all Tanner stages. CONCLUSIONS: BIA appears to be a valid and reliable measure of FFM but is no better than TSF in predictions of body fat. PMID- 9347286 TI - A method to achieve control of dietary macronutrient composition in ad libitum diets consumed by free-living subjects. AB - OBJECTIVE: To validate a shop system in controlling macronutrient composition during ad libitum dietary intervention. DESIGN: Six months randomized intervention trial. SETTING: A shop at the department from which all foods were collected free of charge and registered by a purpose-designed computer system. SUBJECTS: Sixty-five free-living obese subjects (25 kg/m2 < BMI < 34 kg/m2) recruited through advertisement and from a waiting list at the Department. Total drop-out rate was 8%. INTERVENTIONS: Ad libitum low-fat diets (30 energy-% (E%) fat): (1) High-protein (25 E% protein, HP) or (2) Low-protein, (12 E% protein, LP) or habitual diet (controls, C). MAIN OUTCOME MEASURES: Compliance was assessed by 24 h urinary nitrogen excretion (24 h UN). RESULTS: After one month of dietary intervention 24 h UN increased significantly in the HP group and decreased significantly in the LP group (Group difference 95% CI):6.8 g (5.0-8.7 g), P < 0.0001). This group difference remained throughout the trial. There was good agreement between protein intake as estimated by the shop computer and as estimated from 24 h UN in both first (r = 0.86) and second half of the intervention (r = 0.80). CONCLUSION: The high dietary compliance demonstrates the potential of this method to control macronutrient composition in ad libitum dietary intervention studies in free-living subjects. PMID- 9347285 TI - Plasma (carotenoids, retinol, alpha-tocopherol) and tissue (carotenoids) levels after supplementation with beta-carotene in subjects with precancerous and cancerous lesions of sigmoid colon. AB - OBJECTIVES: (1) To compare tissue and plasma carotenoids status of healthy subjects and subjects with pre-cancer and cancer lesions; (2) to evaluate the effect of beta-carotene supplementation on the concentrations of other carotenoids in tissue (luteine + zeaxanthin, cryptoxanthin, lycopene, alpha carotene) and in plasma and also retinol and alpha-tocopherol levels. DESIGN: Eighteen subjects were divided into three groups on the basis of colonoscopy and histological analytical findings: four healthy subjects (control group A); seven subjects affected by adenomatous polyps (group B with pre-cancer lesions); seven subjects suffering from colonic cancer (group C). Blood and colonic biopsy samples were taken (of colon and rectal mucosa) before and after beta-carotene supplementation in all subjects. Groups A and B received a daily dose of beta carotene (30 mg/die) for 43 d. Group C's supplementation was terminated at the time which was performed, usually within 15 d. The tissue and plasma concentration of carotenoids, retinol and alpha-tocopherol were determined by high-performance liquid chromatography. RESULTS: The tissue concentrations of each carotenoid were similar in all the intestinal sites examined as regards groups A and B, although there was a high degree of intra individual variability within each group. Only beta-carotene made significant increases (P < 0.001) after supplementation. The subjects with cancer show tissue levels for each carotenoid lower than those of healthy subjects or subjects with polypous. The plasma levels of alpha-tocopherol did not change after supplementation while significant increases were noted of retinol, alpha-carotene (P < 0.01) and of beta-carotene (P < 0.001). CONCLUSIONS: The patients with colonic cancer seemed to undergo a significant reduction in their antioxidant reserves with respect to the normal subjects and or polyps. We can confirm that oral B-carotene supplementation induces also an increase in plasma alpha-carotene in all groups. PMID- 9347288 TI - Prediction of resting energy needs in older men with heart failure. AB - OBJECTIVES: Patients with congestive heart failure are often undernourished. The measurement of resting energy expenditure has served as the basis upon which estimates of daily caloric needs have been developed. Resting energy needs, however, in heart failure patients are unknown. We have developed a new equation to predict resting energy needs in heart failure patients that takes into account easily measured clinical variables and symptom severity. DESIGN: Observational. SETTING: Baltimore VA Medical Center. SUBJECTS: Forty male patients with heart failure aged, 57-85 y; (27 Class II; 13 Class IV). MEASUREMENTS: Resting metabolic rate was measured by indirect calorimetry, fat-free mass and fat mass by dual energy X-ray absorptiometry, peak VO2 by a treadmill test. Symptom severity was measured by the New York Heart Association classification. Ejection fraction, plasma albumin and plasma glucose were also assessed. RESULTS: Stepwise regression analysis showed that body weight, fasting glucose, plasma albumin and New York Heart Association classification accounted for 83% of the variation in resting energy needs. The regression equation had a root mean square error of 130 kcal (544 kJ) per day. The equation is: RMR (kcal/d) = 12.2 (wt, kg) + 1.6 (glucose, gm/dl) + 103 (NYHA; III, IV)-144 (albumin, mg/dl) + 755. Moreover, prediction equations based on observations in healthy individuals significantly underestimated resting energy needs in heart failure patients. CONCLUSION: We offer a new equation to predict resting energy needs in heart failure patients based upon readily available clinical measurements. Further studies are needed to cross-validate our equation. PMID- 9347289 TI - Supplementation with wine phenolic compounds increases the antioxidant capacity of plasma and vitamin E of low-density lipoprotein without changing the lipoprotein Cu(2+)-oxidizability: possible explanation by phenolic location. AB - OBJECTIVES: To evaluate the effect of the red wine phenolic compound (RWPC) dietary supplementation without alcohol interference on: (1) some of the biochemical characteristics of LDL, (2) the oxidative susceptibility of LDL and (3) the antioxidant capacity of total plasma (Pl-AOC). In order to account for discrepancies between the three series of data, the in vitro stability of the association of phenolic compounds and LDL was tested. DESIGN: An intervention study with 20 volunteers. Each served as his own control. Cu(2+)-oxidizability of LDL and Pl-AOC were tested on blood samples before and after dietary supplementation. Cu(2+)-oxidizability of LDL was also tested by co-incubation in the presence of RWPC or phenolic acids with or without extensive dialysis. SETTING: The Laboratory of Lipid Biochemistry and Biology, School of Medicine, and the Laboratory of Metabolic Diseases, Lapeyronie Hospital, University of Montpellier, France. SUBJECTS: Healthy males, nonsmokers and moderate drinkers, submitted to a dietary regimen deprived of vitamin E and C for a period of 10 d before supplementation. They also abstained from alcohol, wine, fruit juices, coffee, tea and cola beverages during this period. INTERVENTION: Six 0.33 g capsules/d (namely two capsules at each meal) of a preparation of red wine phenolic compounds in a dry powder form were given to the volunteers over a period of two weeks. Blood samples were drawn in fasting conditions at day 0 and day 14 of the supplementation period. RESULTS: Supplementation led to: (1) in LDL, a significant increase in vitamin E content (n = 20, P = 0.01) or vitamin E/total fatty acid bis-allylic carbon number ratio (n = 20, P = 0.006) without modification in the other biochemical characteristics or Cu(2+)-oxidizability; (2) in plasma, a significant increase in the antioxidant capacity (n = 11, P = 0.01). In vitro studies showed that RWPC or sinapic, caffeic or ferulic acids incubated in the presence of LDL increased the protection of the lipoparticle against oxidation (caffeic > sinapic > ferulic). This effect, however, was totally lost after extensive dialysis. CONCLUSIONS: The enhancing effect of the RWPC supplementation on Pl-AOC may be due to a phenolic-compound action both in the aqueous phase of plasma and at the surface of lipoprotein particles. Surface location possibly explains the enhancing-sparing effect of supplementation on LDL vitamin E and the absence of effect on dialysed-LDL oxidizability. PMID- 9347290 TI - Plasma total homocysteine in a representative sample of 972 British men and women aged 65 and over. AB - OBJECTIVES: To provide a reference range for plasma total homocysteine (tHcy), an independent risk factor for vascular disease, and to explore relationships with nutritional indices for people aged 65 y and over, in the UK National Diet and Nutrition Survey (NDNS). DESIGN: The survey procedures described in the National Diet and Nutrition Survey Report (1997) included a health-and-lifestyle interview, a four-day weighed diet record, anthropometric and blood pressure measurements and a fasting blood sample for biochemical indices, including tHcy. SETTING: Eighty randomly selected postcode sectors from mainland Britain during 1995-1996. SUBJECTS: Of 2060 people interviewed, 1527 were visited by the nurse, 1276 gave a blood sample and 972 had tHcy measured. About 80% were in their own homes and the remainder were in nursing homes or similar institutions. RESULTS: Significant cross-sectional relationships, both univariate and multivariate were found between tHcy and index concentrations of folate and vitamin B12 (P < 0.0001), and between tHcy and plasma creatinine, urea, calcium, zinc, alpha 1 antichymotrypsin, lutein and cysteine (P = 0.013 to < 0.0001). Dietary nutrient analyses showed an association with folate intake. tHcy was also correlated with age and with domicile (free-living or institution), with history of vascular disease and with use of four classes of drugs, two of which are prescribed for vascular diseases. There was a north-south gradient in tHcy (P = 0.005), and also in food choices, blood micronutrient indices and vascular disease prevalence. CONCLUSIONS: The concentrations of tHcy found in this study provide a reference range for people aged 65 y and over, in mainland Britain. tHcy is a valuable functional index of micronutrient status and intakes for British people aged 65 y and over, which can assist the development of health-promotion strategies. PMID- 9347291 TI - Vitamin A deficiency among adolescent female garment factory workers in Bangladesh. AB - OBJECTIVE: To investigate the prevalence of vitamin A deficiency among adolescent female factory workers in Bangladesh, and examine the association between various factors and vitamin A status. DESIGN: A cross-sectional study. SETTING: Garment factories in Dhaka city, Bangladesh. SUBJECTS: Three hundred and eighty eight adolescent girls aged 12-19 y from ten garment factories were selected randomly for the present study. Information on socio-economic conditions and usual pattern of dietary intake were obtained by interview. Anthropometric data and blood samples were collected following the interview. RESULTS: By NCHS reference standard, 15.5% of the participants were thin (< 90% Wt/Ht) and about 7% overweight (> 120% Wt/Ht). In about 56%, serum vitamin A level was below the adequate level of 1.05 mumol/l, with 14% having vitamin A deficiency (< 0.70 mumol/l). Forty four per cent of the participants were found to be anaemic (haemoglobin < 120 g/l). Food frequency data on vitamin A rich foods revealed that a large percentage of the participants do not take eggs (41%), milk (64%), liver (85%) and sweet pumpkin (85%); while about 40% of the girls take dark green leafy vegetables (DGLV) and 17% take small fish at least four servings a week. The girls who consumed four or more servings per week of DGLV had significantly higher serum vitamin A level than the girls who took three servings or less. There was a significant positive association between the level of serum vitamin A and frequency of intake of DGLV (r = 0.12; P = 0.023). When age, level of education, percapita income, haemoglobin concentration, serum protein concentration, menstruation at the time of blood collection, prevalence of current morbidity, frequency of intake of egg, milk, small fish, DGLV, liver and sweet pumpkin were accounted for by multiple regression analysis, a strong relationship was found for serum vitamin A concentration with age, menstruation, haemoglobin level and frequency of intake of DGLV. For every unit change in the frequency of consumption of DGLV, there was 0.013 mumol/l change in serum vitamin A level whilst taking other factors into account. CONCLUSION: The data show that there is a high prevalence of subclinical vitamin A deficiency among the adolescent female garment factory workers in Bangladesh, although the anthropometric indices suggest that they do not suffer from acute under nutrition. Consumption of DGLV appears to have an important relation with the vitamin A status of these girls. PMID- 9347292 TI - Age at introduction of complementary food and physical growth from 2 to 9 months in rural Senegal. AB - OBJECTIVE: To compare nutritional status and physical growth among infants according to age at introduction of complementary food (CF). DESIGN: A longitudinal observational study. SETTING: Three health clinics in a rural area of Senegal. SUBJECTS: During immunization sessions, 522 infants were recruited at 2-3 months. Complete data on three visits were available for 420 infants (4 visits: n = 361); 73 were lost to follow-up and 29 had incomplete data. MAIN OUTCOME MEASURES: Increments in length and weight between adjacent visits. RESULTS: Infants complemented at 2-3 months (n = 50) had significantly lower length-for-age (P = 0.014), weight-for-length (P < 0.001) and arm circumference (P < 0.0001) at 2-3 months than predominantly breastfed infants (n = 370), after adjustment for residence, mother's age and education of parents. The growth in weight and length from 2-3 to 9-10 months did not differ. The infants complemented by 4-5 months, but not yet at 2-3 months, (n = 94) had a slightly lower length increment from 4-5 to 6-7 months (1.42 vs 1.53 cm/mo, p < 0.05) compared to infants predominantly breastfed at 4-5 months (n = 276). The infants first complemented by 6-7 months (n = 122) had increments from 6-7 to 9-10 months similar to those of predominantly breastfed infants (n = 154). CONCLUSION: Introduction of CF by 2-3 months was associated with a low nutritional status, but not with slow growth from 2-3 to 9-10 months. Introduction of CF by 4-5 months was associated with slightly slower linear growth compared to later introduction. PMID- 9347293 TI - Using cross-check questions to address the problem of mis-reporting of specific food groups on Food Frequency Questionnaires. UKWCS Steering Group. United Kingdom Women's Cohort Study Steering Group. AB - OBJECTIVE: To explore the potential mis-reporting of specific food groups from food frequency questionnaire (FFQ) data and to examine the effect of using a weighting factor on estimated nutrient intake and ranking of subjects within the cohort according to nutrient intake. DESIGN AND SUBJECTS: A weighting factor was calculated for each of the individual 6572 women aged 35-69 y for four food groups, fish, meat, vegetables and fruit, using FFQ data and cross-check responses. RESULTS: The vegetables weighting had most effect on median intakes, particularly of fibre, vitamins A, C and E and folate. When all the weightings were applied, the median intakes of vitamins A and E were reduced by 35% and 27% respectively and the vitamin C intake was reduced by 44%. Ranking of subjects within the cohort according to nutrient intake was barely affected by the fish and meat weightings. The vegetable weighting had most effect on vitamin A with a rank correlation coefficient of 0.88. When all the weightings were applied the rank correlations for vitamins A, C and E and folate were all less than 0.90. CONCLUSION: Inclusion of cross-check questions in FFQs can provide an additional source of information on food group intake. This can be compared with FFQ data to help identify possible over-reporting and then to adjust frequency of intake accordingly. PMID- 9347294 TI - Study with male volunteers measured 24 h energy expenditure (24 h EE) after consumption of medium-chain (MCT) or long-chain triglycerides (LCT) PMID- 9347295 TI - Paleolithic nutrition. PMID- 9347296 TI - Subtalar arthrodesis with interposition tricortical iliac crest graft for late pain and deformity after calcaneus fracture. AB - We are reporting our experience using tricortical interposition iliac crest grafting in the management of late pain and deformity after calcaneus fracture. Ten patients underwent this procedure, which was performed by the senior author. All but one were followed up with a questionnaire, physical examination, and repeat x-rays. The technique failed in one severely osteoporotic individual because of the graft sinking into the calcaneus. The experience led to two technique modifications that were evaluated in this study. PMID- 9347297 TI - Isolated talocalcaneal interposition fusion: a prospective follow-up study. AB - This is a prospective clinical and radiological study of the treatment of talocalcaneal deformity or degeneration by a modified technique of isolated talocalcaneal fusion. Thirty-six patients were evaluated with clinical examination, plain dorsoplantar and oblique radiographs, and computed tomography scanning or magnetic resonance imaging in a follow-up of 32.5 months (range, 20 62 months). Indications for arthrodesis were posterior tibial tendon rupture with secondary osteoarthritis (12 cases) and secondary posttraumatic osteoarthritis (24 cases). On a visual analog pain scale, the patients graded their pain at 4.4 before surgery and at 1.1 after surgery. The subjective results were 33% complete satisfaction, 28% satisfaction with minor reservation, 31% satisfaction with major reservation, and 9% dissatisfaction. The overall objective results were excellent in 47%, good in 31%, fair in 17%, and poor in 6% of cases. A further advantage of this type of talocalcaneal fusion is a large remaining range of motion in the neighboring joints, at the ankle (in 76% the same or better ROM than before surgery), and at Chopart's joint (in 69% the same or better ROM than before surgery). The fusion rate was high (95%). PMID- 9347298 TI - Biomechanical evaluation of polylactide absorbable screws used for syndesmosis injury repair. AB - Simulated syndesmosis injuries were created in 12 fresh-frozen, below-knee cadaver specimens. Six specimens were repaired with a 4.5 mm stainless steel screw, and six were repaired with a 4.5 mm polylactide screw. Three specimens of each group were tested in load to failure by axially loading with 1400 N and externally rotating to 90 degrees. Three specimens in each group underwent fatigue testing by axially loading with 700 N and applying 2.5 N-m of torque for 57,700 cycles. Radiographs and computed tomography scans were evaluated. None of the screws broke or failed. Similar load to failure was noted in polylactide and control groups. Fatigue testing revealed no significant change in stiffness. No significant screw damage was evident on radiographic or computed tomography evaluation. The data suggest that a polylactide screw has sufficient fatigue and failure strength to allow for healing of this injury in a clinical situation. PMID- 9347299 TI - Treatment of osteochondritis dissecans of the talus: use of the mosaicplasty technique--a preliminary report. AB - A one-stage autogenous osteochondral grafting technique for the treatment of talar dome osteochondritis dissecans is described. Eleven patients with osteochondritis dissecans lesions, 10 mm or greater in diameter, were operated on using the mosaicplasty autogenous osteochondral transplantation technique. Osteochondral cylindrical grafts from the ipsilateral knee were delivered into the talar defect using specially designed tube chisels. These procedures were done by arthrotomy. With follow-up of 12 to 28 months (mean, 16 months), the patients returned to full activities and the results, using the Hannover scoring system, have been excellent. PMID- 9347300 TI - Walking pattern of midfoot and ankle disarticulation amputees. AB - Measurements of the vertical component of ground reaction force (ORF) and dynamic center of pressure (COP) were recorded for five subjects with midfoot level amputations and six with Syme's ankle disarticulation amputations. All of the subjects underwent amputation surgery as a consequence of peripheral vascular disease and diabetes. GRF measurement was accomplished with the F-Scan system (Tekscan, Boston, MA). Each group exhibited a consistent, reproducible pattern of gait. Subjects with Syme's ankle disarticulation initiated initial loading response, i.e., heel strike, with a concentration of GRF in the center of the anatomic heel. COP progressed along the midline to the center of the anatomic forefoot, where GRF was concentrated at push-off. Midfoot amputees initiated loading at the lateral-posterior heel. COP progressed medially to the midline, where it progressed distally to the level of the distal residual limb (proximal metatarsal metaphyses). It then shifted medially under the base of the first metatarsal, where a small concentration of GRF occurred at push-off, similar to the normal foot. These findings explain the decreased magnitude of propulsion seen in midfoot level amputees and may explain the seemingly paradoxical increased metabolic cost of walking observed in midfoot amputees as compared with Syme's ankle disarticulation amputees. PMID- 9347301 TI - A biomechanical comparison of intramedullary nail and crossed lag screw fixation for tibiotalocalcaneal arthrodesis. AB - This study compared the mechanical bending and torsional properties of intramedullary nail fixation and lag screw fixation for tibiotalocalcaneal arthrodesis. Seven matched pairs of human cadaver lower extremities were studied, with one hindfoot in each pair stabilized with a 12 mm x 150 mm interlocked intramedullary nail inserted retrograde across the subtalar and ankle joints. The contralateral hindfoot was stabilized with two crossed 6.5 mm cannulated screws inserted across both the ankle and subtalar joints. Specimens were subjected to cantilever bending tests in plantarflexion, dorsiflexion, inversion, and eversion and to torsional tests in internal and external rotation. The intramedullary nail construct was significantly (P < 0.05) stiffer than the crossed lag screw construct in all four bending directions and both rotational directions: plantarflexion (nail, 42.8 N/mm; screws, 16.4 N/mm; P = 0.0003), dorsiflexion (nail, 43.0 N/mm; screws, 10.3 N/mm; P = 0.0005), inversion (nail, 37.7 N/mm; screws, 12.3 N/mm; P = 0.0024), eversion (nail, 35.4 N/mm; screws, 10.8 N/mm; P = 0.0004), internal rotation (nail, 1.29 N-m/degree; screws, 0.82 N-m/degree; P = 0.01), external rotation (nail, 1.35 N-m/degree; screws, 0.44 N-m/degree; P = 0.0001). Intramedullary fixation is biomechanically stiffer than crossed lag screws in all bending and torsional directions tested and therefore this construct may aid in maintaining alignment of the hindfoot during union and may help increase fusion rate through increased stability of the internal fixation. PMID- 9347302 TI - Stability of the arch of the foot. AB - We defined the relative contributions of six ligaments in stabilizing the arch of the foot: plantar aponeurosis, long-short plantar ligaments, plantar calcaneonavicular ligament (spring ligament), medial talocalcaneal ligament, talocalcaneal interosseous ligament, and tibionavicular portion of the deltoid ligament. Nineteen fresh-frozen human foot specimens were used. A load of 445 N was applied axially to simulate standing-at-ease posture. Three-dimensional positions of tarsal bones before and after ligament sectioning were determined with the use of a magnetic tracking device. The motions were presented in the form of screw axis displacements, quantitating rotation, and axis of rotation orientation. After sectioning one structure, the arch did not collapse on any specimen and there was no obvious change by visual inspection. There were, however, measurable changes in tarsal bone position. Metatarsal-to-talus total rotation difference was greatest with spring ligament and deltoid ligament sectioning, with an average of 2.1 degrees +/- 1.7 degrees and 2.0 degrees +/- 0.2 degree difference, respectively. Calcaneus-to-talus rotation difference was greatest with talocalcaneal interosseous ligament sectioning, with an average of 1.7 degrees +/- 1.5 degrees. The spring ligament, deltoid ligament, and talocalcaneal interosseous ligament were most important for arch stability. PMID- 9347303 TI - Morphology of the plantar calcaneocuboid ligaments. AB - The plantar calcaneocuboid ligaments have various configurations and are referred to in the literature by different names. The aims of this study were to classify the form and attachments of the plantar calcaneocuboid ligaments and to determine their dimensions to describe their morphology. The plantar aspects of 59 cadaver feet were dissected to expose the associated ligaments. The long and short plantar ligaments showed several structural variations of shape, band number, and attachments. The variations that were seen have led to confusion in the literature, which needs to be resolved. A new nomenclature is suggested based on the observed morphology. These findings agree with previous reports, and data are also presented that describe the shapes of the ligaments. PMID- 9347304 TI - Valgus stress radiography in normal ankles. AB - Deltoid ligament injury is thought to be rare. Signs of complete rupture of the deltoid ligament may be subtle or interpreted as another injury condition and thus are often missed acutely. No standardized method has been created to evaluate medial ligament insufficiency in acute or chronic laxity. To establish a diagnostic test for suspected isolated ruptures of the deltoid ligament, 32 subjects with no previous ankle injury underwent valgus stress radiography and nonstressed radiography of both ankles. Stress radiography in this study showed that there is a measurable but minimal range of talar tilt on valgus stress in previously uninjured ankles. This study provides the basis for diagnosis of the rare isolated rupture of the deltoid ligament of the ankle. PMID- 9347305 TI - Posterior tibial tendon dysfunction: secondary MR signs. AB - We evaluated four potential secondary magnetic resonance imaging signs to aid in clinical diagnosis of posterior tibial tendon (PTT) tears. Seventy-one ankles (25 PTT tears and 46 controls) were evaluated for the following secondary signs: (1) PTT sheath fluid, (2) a distal tibial spur located just anterior to the PTT, (3) unroofing of the talus, and (4) "bone bruise"--like medullary lesions. Two musculoskeletal radiologists rated their confidence using a scale and were compared for level of agreement. The presence of PTT sheath fluid had modest specificity and fair to moderate sensitivity. Tibial spurring and unroofing of the talus had excellent specificity and fair sensitivity. Bone bruise-like lesions were commonly seen in cases and controls. Examination of divergence of opinion between the two radiologists revealed pitfalls in interpretation of PTT sheath fluid and bone bruise-like lesions, which were commonly the result of adjacent vessels and inhomogeneous fat saturation, respectively. We conclude that secondary signs of PTT tears with high specificities include unroofing of the talus, tibial spurring, and PTT sheath fluid. PMID- 9347306 TI - Complete posterior dislocation of the talus: a case report. PMID- 9347307 TI - Complex interphalangeal dislocation of the fourth toe after a sports injury: case report. PMID- 9347308 TI - Glomus tumor of the plantar arch: a case report with magnetic resonance imaging findings. AB - A 55-year-old woman with a 12-year history of a painful nodule in the subcutaneous fat layer of the plantar arch was evaluated with magnetic resonance imaging, followed by excisional biopsy. Pathology revealed a glomus tumor, which is extremely rare in the plantar surface of the foot. The magnetic resonance imaging studies are presented. The literature on glomus tumors in the foot is reviewed. This entity should be considered in the differential diagnosis of solitary plantar nodules when marked sensitivity to temperature or pressure is exhibited. PMID- 9347309 TI - Treatment of diabetic foot ulcers and Charcot neuroarthropathy using the patellar tendon-bearing brace. PMID- 9347310 TI - A new operative approach for flatfoot secondary to posterior tibial tendon insufficiency: a preliminary report. PMID- 9347311 TI - Angiotensin II receptors in endothelial cells. AB - 1. Angiotensin II (Ang II), the main effector of the renin-angiotensin system, exerts its vasoconstrictory and trophic actions on smooth muscle cells via AT1 receptors. However, Ang II does not act only on smooth muscle cells, as Ang II receptors are also present in endothelial cells. 2. The receptor type on these cells differs depending on the origin of the endothelium and the species. The rat endothelial receptors are mostly of the AT1 type, but AT2 receptors have also been found. The pharmacological characteristics of the AT1 receptors on endothelial cells are similar to those of other cell types. 3. Ang II stimulates phospholipase C and phospholipase A2 activation via the AT1 receptor in endothelial cells. Ang II also stimulates the tyrosine phosphorylation of several proteins in these cells. 4. Some studies suggest that the AT1 receptor mediates the release of vasodilator molecules by endothelial cells and could modulate Ang II effect on smooth muscle cells. Ang II may also inhibit endothelial cell growth via the AT2 receptor. Finally, endothelial Ang II receptors may be implicated in the regulation of fibrinolysis. PMID- 9347312 TI - Inhibition of nitric oxide formation by aminoguanidine: an attempt to prevent insulin-dependent diabetes mellitus. AB - 1. Insulin-dependent diabetes mellitus is an autoimmune disease leading to pancreatic beta-cell destruction, an event that may, at least partially, be induced by the formation of nitric oxide. 2. Under the influence of cytokines, the enzyme nitric oxide synthase is induced. 3. Blockage of the inducible form of nitric oxide synthase has been found to protect against insulin-dependent diabetes mellitus in some animal models. 4. Aminoguanidine has been found to be a fairly specific inhibitor of cytokine-inducible nitric oxide synthase. 5. Aminoguanidine may reduce the blood flow to the pancreatic islets in vivo and, at higher concentrations, also impair insulin secretion by the beta-cells,--which may make the compound less useful in attempts to prevent insulin-dependent diabetes mellitus. PMID- 9347313 TI - Antitumor activity of 2,2'-bipyridyl-6-carbothioamide: a ribonucleotide reductase inhibitor. AB - 1. 2,2'-Bipyridyl-6-carbothioamide (BPYTA) is a synthesized compound with chelating properties demonstrating in vitro and in vivo antitumor activity. 2. The BPYTA cytotoxic effect is mainly due to the inhibition of ribonucleotide reductase (RR), a key enzyme in proliferating cells. The active form of BPYTA is its iron chelate [BPYTA-Fe(II), molar ratio 2:1], which destroys the tyrosyl radical of RR small subunit (R2). 3. The copper chelate of BPYTA [BPYTA-Cu(II), molar ratio 2:1] also has antiproliferative activity, but RR is not the only intracellular target. 4. BPYTA potently synergizes in vitro with hydroxyurea, the most widely used R2 inhibitor. PMID- 9347314 TI - Increased contractile response induced with ouabain is abolished by thapsigargin in aorta of renal hypertensive rats. AB - 1. The aim of the present study was to test in vitro if the increased contractile effect of phenylephrine and KCl, observed after the addition of ouabain in renal hypertensive rat aorta, is mediated by Ca2+ accumulated on the sarcoplasmic reticulum. 2. In aortas of one kidney (1K) rats, ouabain did not modify the concentration-effect curves stimulated with phenylephrine and KCl. 3. Contractile responses stimulated with phenylephrine and KCl were potentiated by ouabain in one kidney--one clip (1K-1C) aortas, and preincubation with thapsigargin abolished the increasing effect of ouabain on these contractions. 4. The addition of thapsigargin before phenylephrine and KCl did not modify the contractile response to phenylephrine and KCl or the resting vascular tone during the time incubation. 5. In the presence of ouabain, thapsigargin significantly increased the vascular tone only in 1K-1C rat aortas. 6. Increased intracellular Na+ concentration as a consequence of Na(+)-K(+)-ATPase inhibition induces increased accumulation of Ca2+ inside the sarcoplasmic reticulum in 1K-1C rat aortas. The differential effects in renal hypertensive and normotensive aortas suggest a possible role of this mechanism in modulating cytosolic Ca2+ in renal hypertension. PMID- 9347315 TI - Irritant action of monochloramine in rat stomachs: effects of zinc L-carnosine (polaprezinc). AB - 1. Effects of a novel zinc compound (polaprezinc), N-(3-aminopropionyl)-L histidinato zinc, on the mucosal ulcerogenic response induced by ammonia (NH4OH) and monochloramine (NH2Cl) were examined in rat stomachs. 2. Oral administration (1 ml) of NH4OH (> 600 mM) and NH2Cl (> 60 mM) produced severe hemorrhagic lesions in unanesthetized rat stomachs, whereas hypochlorous acid (HClO) even at 120 mM did not cause any macroscopic damage. 3. Pretreatment of the animals with polaprezinc (2-12 mg/ml, 1 ml, PO) showed a dose-dependent inhibition against gastric lesions induced by NH4OH (1,800 nM) or NH2Cl (120mM), and this effect was significant at > 6 mg/ml in either case. These lesions were also significantly prevented by prior administration of dmPGE2 (2 micrograms/ml, 1 ml, PO). 4. Mucosal application of NH4OH (300 mM) and NH2Cl (10 mM) caused a marked reduction of transmucosal potential difference (PD) in ex vivo stomachs of anesthetized rats. The reduced PD responses caused by NH4OH and NH2Cl were prevented dose dependently by preexposure of the mucosa to polaprezinc, but not affected by dmPGE2. 5. Mucosal exposure to NH4OH (60 mM) caused a marked PD reduction in ex vivo stomachs made ischemic by bleeding from the carotid artery (1 ml per 100 g body wt), followed by severe gastric lesions. These ulcerogenic and PD responses caused by NH4OH plus ischemia were attenuated by prior application of polaprezinc as well as taurine (25 mg/ml, 1 ml), while dmPGE2 prevented the lesions without affecting the reduced PD response. 6. These results suggest that (a) NH2Cl damages the gastric mucosa at much lower concentrations than NH4OH, (b) polaprezinc protects the stomach against injury caused by either NH2Cl or NH4OH, and (c) the mechanisms underlying the protective action of polaprezinc remain unclear but may be different from those of dmPGE2. PMID- 9347316 TI - Effects of chronic and acute ethanol exposure on renal (Na + K)-ATPase in the rat. AB - 1. We evaluated the effects of chronic ethanol consumption on the kinetic properties of renal (Na + K)-ATPase and compared them with acute inhibition by ethanol in vitro. 2. When adult rats were fed 20% ethanol for 10 weeks, renal (Na + K)-ATPase activity increased but the sensitivity of the enzyme to ethanol inhibition in vitro was not altered. 3. Vmax was increased by ethanol consumption, whereas K0.5 and nH were not changed. The kinetic parameters of Mg(2+)-ATPase were not affected under the same conditions. 4. We concluded that ethanol-induced tolerance or enhancement of renal (Na + K)-ATPase or both can be explained on the basis of an increase in Vmax. PMID- 9347317 TI - Acetate and propionate potentiate the antiproliferative effect of butyrate on RBL 2H3 growth. AB - 1. The effect of acetate, propionate, and butyrate separately and combined on RBL 2H3 (a rat basophilic leukemic cell type) proliferation during 24, 48, and 72 hr was examined. Also, the effect of a mixture of the three volatile fatty acids on proliferation of HeLa-155 (a human adenocarcinoma), C57 B1/6J (a mouse melanoma), and MCF-7 (human breast tumor) during 8 days was investigated. 2. Acetate and propionate per se did not present any effect on RBL-2H3 growth during 72 hr, however, when acetate and propionate were added together a significant inhibition of this cell growth was found; 18% for 48 and 37% for 72 hr. The addition of butyrate to the culture medium caused a 75% decrease in the rate of this cell growth either after 48 and 72 hr. This effect of butyrate was pronounced by acetate (86% and 90% for 48 and 72 hr, respectively), propionate (87% for 48 and 93% for 72 hr), and acetate and propionate together (76% for 48 and 92% for 72 hr). 3. Daily addition of a mixture of the short-chain fatty acids (10 mM acetate, 2 mM propionate and 1.5 mM butyrate) markedly decreased the number of cells after 8 days: 58% for RBL-2H3, 42% for HeLa-155, 91% for C57 B1/6J and 55% for MCF-7. 4. These results support the proposition that a fiber-rich diet that leads to great production of butyrate but also of propionate and acetate would be more effective to prevent the occurrence of colorectal cancer than the administration of this short-chain fatty acid given alone. PMID- 9347318 TI - Antagonistic effects of isofloxythepin on dopamine D1 and D2 receptors and behaviors in rats. AB - 1. In vitro, isofloxythepin competed for the binding of [3H]SCH 23390 to striatal D1 receptors and for the binding of [3H]spiperone to striatal D2 receptors with IC50 values of 6.1 (+/- 1.2) x 10(-10)M and 8.4 (+/- 2.6) x 10(-10)M, respectively. Isofloxythepin further inhibited the D1 receptor-mediated, dopamine stimulated adenylate cyclase in the striatal tissue. 2. Fifteen hours after rats were injected with a single dose of isofloxythepin (1 mg/kg, SC), the ex vivo binding curve of [3H]spiperone to striatal D2 receptors was markedly inhibited, whereas the binding curve of [3H]SCH 23390 to striatal D1 receptors was unaffected. 3. Fifteen hours after isofloxythepin pretreatment, D1 agonist SKF 38393 (15 mg/kg, IP)-stimulated grooming behavior was not affected, whereas the D2 agonist quinpirole (3 mg/kg, IP)-stimulated stereotyped behavior was completely abolished. 4. On the basis of the findings from in vivo studies, we conclude that, although isofloxythepin is found to have high affinity for both D1 and D2 receptors in vitro, it lacks D1 antagonistic potency in vivo. PMID- 9347320 TI - Competitive and silent antagonism of recombinant 5-HT1B receptors by amiloride. AB - 1. The cyclic adenosine monophosphate (cAMP) response of the diuretic amiloride was compared with sumatriptan at recombinant human 5-HT1B (h5-HT1B) receptor sites in stably transfected Chinese hamster ovary (CHO-K1) cells. 2. Amiloride, free of intrinsic activity (pEC50 < 3.0), competitively antagonized the sumatriptan-mediated inhibition (pEC50: 7.37) of 100 microM forskolin-induced cAMP formation with a pA2 value of 4.46. 3. The antagonist feature was not shared by the amiloride derivative ethylisopropylamiloride, notwithstanding a slightly higher 5-HT1B receptor-binding affinity (pKi: 4.87) than that of amiloride (pKi: 4.70). PMID- 9347319 TI - The human 5-HT1A receptor: comparison of its binding properties in transfected cells and cortical tissue. AB - 1. The binding characteristics of tritium labeled 8-hydroxy-dipropyl aminotetralin, or [3H]8-OH-DPAT, to the serotonin1A (5-HT1A) receptor in the stably transfected HeLa cell clone HA6 and in human cortical tissue were examined and compared. 2. A series of kinetic studies of [3H]8-OH-DPAT binding to the transfected HA6 cell line demonstrated two components in both the association and the dissociation reactions. 3. In saturation experiments, at least two affinity states were unequivocally detected in the HA6 cell line and the human cortical tissue. Using isotopic dilutions, the binding isotherms were best fitted to a two site model, and similar affinity values were obtained in both systems (KH approximately 1.1 nM and KL approximately 12-223 nM). 4. Most of the drugs used in competitions inhibited [3H]8-OH-DPAT binding, following a two-site model, and maintained their rank order of binding potency in both systems; that is, 5-HT > or = 8-OH-DPAT > buspirone > pindolol. Inconsistencies, however, were found for the antagonists NAN-190 and pindolol; only one inhibition constant was determined for HA6 cells, but two affinities were detected with cortical tissue. 5. The results indicate that, although data from binding studies using the cell expression system reflect, to a certain extent, those obtained with the cortical tissue, some discrepancies remained. 6. Finally, and in contrast with what is observed with the 5-HT1A receptor expressed in the HA6 cell line, it is possible that different receptors, or subtypes of one receptor, or even uptake sites normally expressed in cortical tissue, could interact with [3H]8-OH-DPAT or the competing drugs or both, thus leading to the observation of additional binding sites. PMID- 9347321 TI - Inhibition of muscarinic receptor-operated Ca2+ sensitization by short-term desensitization of alpha-toxin-permeabilized smooth muscle cells from guinea pig stomach. AB - 1. Isolated single smooth muscle cells from the guinea pig stomach were permeabilized with Staphylococcus aureus alpha-toxin. 2. The permeabilized single cells showed a shortening in response to Ca2+ in an all-or-none manner. Moreover, the addition of acetylcholine (ACh) or guanosine 5'-triphosphate (GTP) resulted in a decrease in concentration of Ca2+ required to trigger a threshold response, suggesting that Ca2+ sensitization is induced by the stimulation of muscarinic acetylcholine receptors (mAChRs) or GTP-binding protein(s). 3. Short-term desensitization was induced by incubating the permeabilized cells with 100 microM ACh for 10 min. 4. In desensitized cells, the concentration of Ca2+ required to trigger a threshold response in the presence of ACh was increased, however, the cell shortening in response to Ca2+ in the absence of ACh and GTP-induced Ca2+ sensitization was not affected by short-term desensitization. 5. These results suggest that the receptor-operated augmentation of Ca2+ sensitivity is inhibited by short-term desensitization and that the development of short-term desensitization is due to an uncoupling of mAChR/GTP-binding protein(s). PMID- 9347322 TI - A difference in responsiveness of isolated hepatic artery, aorta, portal vein and vena cava of pig to flavin adenine dinucleotide including liver extract. AB - 1. Tension of isolated hepatic artery, aorta, portal vein and vena cava of pigs was measured isometrically to study the mode of action of flavin adenine dinucleotide, including liver extract (FADLE). 2. FADLE showed concentration dependent relaxation of norepinephrine contraction in the hepatic artery, but FADLE had little influence on the aorta, portal vein and vena cava contracted by norepinephrine (NE). 3. In Ca(2+)-free solution, FADLE-induced relaxation in both the hepatic artery and aorta contracted by NE was significantly reduced, and its reduction was greater in the hepatic artery than in the aorta. Furthermore, extracellular Ca2+ dependence of norepinephrine-induced contraction was much larger in the hepatic artery than in the aorta. 4. Results suggest that FADLE shows regional variations in vasorelaxation through the alteration of Ca2+ movement by the cell, and that this may reflect one of the mechanisms in its ability to increase blood flow in hepatic tissue. PMID- 9347323 TI - Role of nitric oxide in the convulsive seizures induced by fluoroquinolones coadministered with 4-biphenyl acetic acid. AB - 1. Contribution of nitric oxide to the convulsive seizures induced by fluoroquinolones (FQs) coadministered with 4-biphenyl acetic acid (BPAA), the active metabolite of fenbufen, was assessed in mice. 2. Enoxacin + 4-biphenyl acetic acid caused clonic seizures in all treated mice, followed by tonic seizures and death. These events were associated with a significant increase in intracerebellar cyclic GMP. 3. Pretreatment with the nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine methylester (L-NAME), but not with D-NAME, significantly reduced the incidence of convulsions and lethality, as well as the increase in cyclic GMP. 4. Pretreatment with N-methyl-D-aspartic acid (NMDA) receptor antagonist, MK-801, inhibited only the transition of clonic seizure to tonic seizure without affecting the incidence of clonic seizure and lethality. 5. These findings suggest that FQs + BPAA exert convulsions by activating NOS partly through the mediation of the NMDA receptor in the brain cells. PMID- 9347324 TI - Anticholinergic and calcium antagonistic activities of NS-21 contribute to the inhibition of rat urinary bladder contractions. AB - 1. Pharmacological characteristics of NS-21 and its main metabolite, RCC-36, in the inhibition of rat urinary bladder contractions were investigated both in vitro and in vivo. 2. NS-21 and RCC-36 inhibited muscarinic receptor bindings to rat bladder membranes with [3H]3-quinuclidinyl bezilate (pKi 6.19 and 7.24, respectively), whereas they failed to inhibit the following receptor bindings: adrenergic (alpha 1, alpha 2 and beta), dopaminergic (D1 and D2), adenosine (A1 and A2), histaminergic (H1) and opioid (mu, delta and kappa) receptors. 3. NS-21 and RCC-36 suppressed carbachol-induced contractions of isolated rat detrusor strips in competitive (pA2 6.80 and 7.93, respectively) and noncompetitive (pD'2 5.93 and 5.77, respectively) manners. 4. NS-21 and RCC-36 inhibited CaCl2-induced contractions of rat detrusor strips in the presence of 100 mM K+ (pIC50 6.54 and 5.76, respectively), as well as the 100 mM K(+)-induced 45Ca influx into the isolated bladder strips at > or = 1 microM. 5. Electrical stimulation of the peripheral end of the pelvic nerve in anesthetized rats induced bladder contractions composed of two phases: an initial phasic contraction that was weakly suppressed by atropine, and a tonic contraction that was strongly suppressed by atropine. NS-21 suppressed both contractions to the same degree (ID50 2.65 and 2.19 mg/kg, i.v., respectively). RCC-36 suppressed the tonic contraction (ID50 1.20 mg/kg, i.v.) more markedly than the initial contraction (ID50 7.43 mg/kg, i.v.). 6. These results suggest that NS-21 and RCC-36 suppressed bladder contractions owing to their anticholinergic and calcium antagonistic activities. PMID- 9347326 TI - Endogenous thromboxane A2 does not contribute to the contractile response of human umbilical artery strips to 5-hydroxytryptamine. AB - 1. To investigate the possible role of endogenous thromboxane A2 (TXA2) in 5 hydroxytryptamine (5-HT)-induced contraction, human umbilical artery strips were suspended in isolated organ chambers for measurement of isometric force. 2. In endothelium intact strips, arachidonic acid (AA;1 microM) potentiates the contractile response to 5-HT, whereas the response was reduced by indomethacin (INDO;10 microM). De-endothelialized strips showed reduced responses to 5-HT. 3. Arachidonic acid-induced potentiation of the responses to 5-HT was prevented by INDO, and the TXA2 synthase inhibitor dazoxiben (DAZ;1 microM and 10 microM) was without effect on the responses to 5-HT in endothelium intact strips. 4. Taken collectively, these results suggest that, in human umbilical artery strips, the contractile response to 5-HT is at least partly dependent on the 5-HT induced release of an endothelium-derived contracting factor (EDCF), which is a cyclooxygenase metabolite. The lack of effect of DAZ indicates that TXA2 is not the EDCF released during the contractile response of human umbilical artery strips to 5-HT. PMID- 9347325 TI - Effect of combined neonatal imprinting by vitamin A, vitamin D3, benzpyrene and allylestrenol on adult rat thymus glucocorticoid and uterine estrogen receptors. AB - 1. Combined neonatal imprinting with allylestrenol, vitamins A and D3 and benzpyrene significantly increased thymic glucocorticoid receptor capacity in male and female animals and decreased receptors affinity in adult females only. 2. Uterine estrogen receptor affinity or density was not influenced. 3. Considering that perinatal treatment with allylestrenol or vitamin D3 decreased glucocorticoid receptor capacity, the dominance of the positive effect of retinol should be surmised. 4. The experiments call attention to the interrelation of different materials acting simultaneously in the perinatal period. PMID- 9347327 TI - Effects of a lipoxygenase inhibitor on digoxin-induced cardiac arrhythmias in the isolated perfused guinea-pig heart. AB - 1. The effects of a lipoxygenase inhibitor, BW A4C, on digoxin-induced arrhythmias and cardiac dynamics (contractile force, perfusion pressure, heart rate) were investigated in Langendorff-perfused isolated guinea-pig hearts. In the control group, arrhythmias were induced by 25 micrograms/ml digoxin at a perfusion rate of 0.5 ml/min. In the treated groups, BW A4C (1 and 0.3 microM) perfused continuously from 15 min prior to digoxin until cardiac arrest occurred. Digoxin exposure (microgram/g wet weight of heart) for the occurrence of arrhythmias and cardiac arrest were the parameters evaluated to assess cardiotoxicity. 2. Digoxin caused a marked increase in leukotriene B4 release in the coronary effluent, and was collected during tachyarrhythmias. BW A4C markedly inhibited the digoxin-induced elevation of LTB4. 3. BW A4C (1 and 0.3 microM) did not prevent the onset of ventricular fibrillation and ventricular tachycardia despite a slight delay in the occurrence of ventricular fibrillation and cardiac arrest at the 0.3 microM concentration. 4. Contractile force increased significantly after digoxin infusion which was concomitant with the time of onset of arrhythmias. In the presence of BW A4C, the contractile force increased, but not significantly. Perfusion pressure increased initially after digoxin infusion in the absence and the presence of BW A4C, but not significantly. 5. These findings show that the lipoxygenase inhibitor lacked any protective action on digoxin-induced arrhythmias despite its effective suppression of digoxin-induced elevation of LTB4 in coronary effluent. PMID- 9347328 TI - Effect of morphine on rotational behavior of unilaterally brain-lesioned mice and rats: morphine withdrawal and development of tolerance. AB - 1. The effect of acute morphine administration on the rotational behavior of unilaterally brain-lesioned mice and on the amphetamine-induced rotational behavior of unilaterally brain-lesioned rats was studied. 2. The effect of repeated morphine administration on the rotational behavior of mice and rats was also investigated. 3. Acute administration of 40 mg/kg of morphine induced strong ipsilateral rotation in unilaterally brain-lesioned mice. 4. One day after withdrawal, mice given morphine repeatedly for 5 days and treated acutely with 40 mg/kg of morphine rotated significantly less ipsilaterally than mice that had received the same dose of morphine for the first time. 5. Rats given 2 mg/kg of morphine 1 hr before administration of 5 mg/kg of amphetamine tended to rotate slightly more in the ipsilateral direction than the similarly lesioned control rats that received amphetamine alone. 6. After 1 day of withdrawal from 5 days of repeated morphine administration, rats given morphine before amphetamine tended to rotate less ipsilaterally than those given morphine before amphetamine for the first time. 7. Thus, repeated administration of morphine appears to induce tolerance to the effect of morphine on circling behavior in unilaterally brain lesioned mice and rats. PMID- 9347330 TI - Twitch potentiation induced by caffeine in the mouse diaphragm depends on external calcium ions in the absence of potassium ions. AB - 1. Removal of external K+ ions increases the amplitude of directly elicited twitch contractions of the mouse diaphragm (Nishimura et al., 1996). This increase depends on external Ca2+ ions. 2. We examined the effect of caffeine (2 mM) on this increase in twitch amplitude. The mouse diaphragm muscle was directly stimulated in the presence of d-tubocurarine (10 microM). 3. Caffeine increased the amplitude of twitches in a standard bathing solution. This effect was maintained in a solution without either K+ or Ca2+ ions but was abolished in a solution from which both ions were absent. Readdition of Ca2+ ions restored the potentiating effect of caffeine. 4. In the presence of caffeine, removal of both K+ and Ca2+ ions decreased the resting membrane potentials of muscle fibers to about -53 mV. The readdition of 2 mM Ca2+ ions restored the membrane potentials. 5. Twitch potentiation in the absence of external K+ ions was attenuated by 10 microM bepridil but not by 3 microM verapamil or 10 microM Cd2+ ions. 6. These results support the hypothesis that Na(+)-Ca2+ exchange can support twitch contraction during the inhibition of Na(+)-K(+)-ATPase activity. The influx of Ca2+ ions into the cells might be stored in the sarcoplasmic reticulum. PMID- 9347329 TI - Effects of feeding conditions on sensitivity to tubocurarine of nerve-muscle preparations from the mouse. AB - 1. The effects of feeding conditions on the sensitivity to the effect of tubocurarine (dTc) in vitro were compared among various nerve-muscle preparations from mice. The mice were fed under conditions that restricted or compelled their movement for 64 days and controls were fed conventionally. 2. The sensitivity to the effect of dTc differed considerably among preparations. It was much higher in the sciatic nerve-extensor digitorum longus muscle (EDL), moderately higher in the sciatic nerve-soleus muscle (SOL) and lower in the phrenic nerve-diaphragm (DPH) in control mice. 3. The order of the sensitivity was not altered by either type of conditioning. Constant restriction of movement or compelled movement did not modify the sensitivity of DPH to the effect of dTc in vitro. 4. Compulsion facilitated the sensitivity in both SOL and EDL. Restriction selectively increased the sensitivity of EDL. Both types of conditioning selectively and significantly reduced twitch development in EDL. 5. These results indicate that the sensitivity to dTc of neuromuscular transmission reflects constant states of motor activity. PMID- 9347331 TI - Effects of feeding condition after birth on the sensitivity of neuromuscular transmission to d-tubocurarine in vitro in mice. AB - 1. Effects of feeding condition from birth were examined on the sensitivity of neuromuscular transmission to d-tubocurarine (dTc) in vitro in male mice of the ddY strain. 2. Mice were trained to climb two separated cylindrical steel-wire tubes for feeding and drinking, respectively, from 16 days of age. Some mice were conventionally fed, from 99 days of age. Nerve-muscle preparations were made from the left phrenic nerve diaphragm muscle (DPH), the sciatic nerve soleus muscle (SOL), and the sciatic nerve extensor digitorum longus muscle (EDL) of 99-day-old and 155-day-old mice. The nerve trunk was electrically activated with trains of four pulses and tetanic pulses. 3. The sensitivity to the effects of dTc decreased in the order EDL, SOL, and DPH. This result held true in all mice tested. 4. This sensitivity was significantly potentiated by the compulsory movement. 5. The supersensitivity remained even when mice were conventionally fed after 99 days of age. 6. The compulsion rendered EDL antifatigable on tetanic stimulation. This property was also retained after a return to conventional feeding. 7. These results suggest that the effects of feeding condition from birth might remain on neuromuscular functions after termination of the conditioning. PMID- 9347333 TI - Actions of serotonergic agents on hypothalamic-pituitary-adrenal axis activity in the rat. AB - 1. The effects of ipsapirone, nefazodone, tiaspirone, BMS-20661, buspirone and gepirone on the hypothalamic-pituitary-adrenal (HPA) axis were studied. These drugs were selected because they have serontonin 1A (5-HT1A) receptor-binding capability and have the potential for therapeutic activity in the treatment of major affective or anxiety disorders or both. 2. Plasma corticosterone level was used as the end point for determining the effect of each drug on the HPA axis. Each drug increased the plasma corticosterone levels in a dose-dependent manner. The ED50 values were 0.8 mg/kg for BMS-20661, 3.5 mg/kg for gepirone, 3.9 mg/kg for buspirone, 5.3 mg/ kg for tiaspirone, 10.5 mg/kg for ipsapirone and 73.5 mg/kg for nefazodone. Ipsapirone and buspirone were more efficacious than the other four drugs. 3. The effect of a 10-mg/kg (35 mg/kg for nefazodone) test dose of each drug reached a peak between 30 min and 1 hr, and plasma corticosterone levels generally returned to control levels after 2 hr. 4. When the drugs were given 30 min before decapitation, in conjunction with a rotatory stress, BMS 20661 significantly inhibited the stress-induced rise, whereas ipsapirone and gepirone caused a significant increase in plasma corticosterone levels. However, when the drugs were given 2 hr before decapitation, nefazodone caused a significant decrease, whereas ipsapirone, BMS-20661 and gepirone produced significant increases in HPA axis activity. An 0800 hr dose of 0.1 mg/kg of dexamethasone suppressed the 1500 hr HPA activity by 73.1%. The 0.1-mg/kg dose of dexamethasone significantly reduced the drug-activated HPA axis activity of all of the drugs from their saline-control levels. The rank order, from least to greatest inhibitory effect, produced by this dexamethasone treatment on the drug control levels was gepirone (-42.6%), tiaspirone (-48.9%), buspirone (-56.1%), nefazodone (-68.5%), insapirone (-70.0%), and BMS-20661 (-74.3%). PMID- 9347332 TI - Nonadrenergic, noncholinergic inhibitory innervation in the longitudinal muscle of the rabbit portal vein. AB - 1. We examined whether nitric oxide (NO) and ATP take part in inhibitory nonadrenergic, noncholinergic (NANC) neurotransmission in the portal vein of Japanese White rabbits. Longitudinal strips of the vein were suspended in organ baths containing Krebs bicarbonate solution. Preparations relaxed in response to electrical field stimulation (EFS) (25 V, 0.5 msec in duration, 1-50 Hz) and exogenous ATP (1-300 microM) after contraction was induced with 10 microM ergotamine in the presence of 3 microM guanethidine and 0.1 microM atropine. 2. The relaxation response to EFS was abolished by addition of 100 microM NG-nitro-L arginine, an NO synthase inhibitor. This abolition was partly reversed by 10 mM L arginine, a substrate for NO synthase. 3. The relaxation response to exogenous ATP was significantly inhibited by 1 mM suramin, a P2X- and P2Y-purinoceptor antagonist, whereas relaxation response to EFS was not inhibited by 1 mM suramin. 4. From the results of this study, we conclude that the inhibitory NANC neurotransmission is mediated by NO alone and does not involve ATP in the portal vein of Japanese White rabbits under our experimental conditions. PMID- 9347335 TI - Amitriptyline: a potent inhibitor of butyrylcholinesterase from human serum. AB - 1. The effect of amitriptyline on human serum butyrylcholinesterase (acylcholine acylhydrolase E.C.3.1.1.8) has been investigated. From the Lineweaver-Burk plot and the plot of v versus amitriptyline concentration, it was concluded that amitriptyline inhibition is partially competitive, and the kinetic parameters have been calculated as Ks = 0.11 mM, alpha = 1425 and Ki = 0.01 mM. 2. Because amitriptyline is a partial competitive inhibitor of butyrylcholinesterase, acquired deficiency may be seen in patients treated with amitriptyline and may cause complications in operations. PMID- 9347334 TI - Relaxin as a relaxant of the isolated rat uterus: comparison with its mechanism of action in vivo. AB - 1. Glibenclamide, a blocker of ATP-sensitive potassium channels, has been shown to antagonize relaxin as a uterine relaxant in the rat in vivo but not in vitro. The aim, therefore, was to investigate whether the discrepancy between the two studies was a consequence of differences in (1) muscle layers, (2) hormonal conditions or (3) spasmogens utilized. Relaxin was compared with salbutamol and levcromakalim. 2. Relaxin was of similar potency as a uterine relaxant against oxytocin (0.2 mM)-induced spasm with tension measured in the circular or longitudinal muscle layers. Glibenclamide (10 microM) did not antagonize relaxin or salbutamol in these preparations but greatly antagonized levcromakalim (91 fold). Relaxin was a relaxant of tension activated by transmural electrical stimulation in uteri from rats that had been ovariectomized, although the maximal effect was only 30 +/- 15%, and in uteri from rats that had been treated with 17 beta-estradiol benzoate. Glibenclamide was not an antagonist of relaxin in the latter preparation but did antagonize levcromakalim (118-fold). Relaxin also inhibited spontaneous phasic tension development in uteri from ovariectomized rats but again was not antagonized by glibenclamide. 3. Because relaxin was not antagonized by glibenclamide under any of these various conditions, it would appear that the in vivo-in vitro discrepancy in the antagonism of relaxin by glibenclamide is not attributable to the effects of different muscle layers, hormonal conditions or spasmogens. It may be that the mechanism of action of relaxin or glibenclamide or both differs between in vivo and in vitro preparations. PMID- 9347336 TI - Analgesic effects of amlodipine and its interaction with morphine and ketorolac induced analgesia. AB - 1. The antinociceptive effects of amlodipine, administered subcutaneously (s.c.), intracerebroventricularly (i.c.v.) and intrathecally (i.t.) were examined with the acetic acid writhing and tail-flick tests in mice. Amlodipine was also tested in combination with morphine and ketorolac. Isobolographic analyses were used to define the nature of functional interactions between amlodipine and morphine or ketorolac. 2. The s.c. (0.1, 1.25, 2.5, 5 and 10 mg/kg), i.c.v. (2.5, 5, 10 and 20 micrograms/mice) and i.t. (2.5, 5, 10 and 20 micrograms/mice) administration of amlodipine exhibited a dose-dependent antinociceptive effect in the writhing test but had no effect on the tail-flick latency. Isobolographic analyses revealed an additive interaction between amlodipine and morphine or ketorolac in the writhing test. 3. These results suggest that amlodipine induces antinociception and increases antinociceptive action of morphine and ketorolac, possibly through a decrease in cellular calcium availability. PMID- 9347337 TI - Depolarization-dependent effect of flavonoids in rat uterine smooth muscle contraction elicited by CaCl2. AB - 1. The effects of the flavonoids genistein (3-60 microM), kaempferol (3-60 microM) and quercetin (1-100 microM) on KCl (60 mM)-induced tonic contraction in rat uterus and their modifications with the inhibitor of cAMP-dependent protein kinases (TPCK, 3 microM), the inhibitor of ornithine decarboxylase [alpha difluoromethyl ornithine (DFMO), 10 mM] and the polyamine spermine (1 mM) have been assayed. The effects of the three flavonoids were also studied on the contraction elicited by CaCl2 (30 microM to 10 mM) on rat uterus incubated in medium lacking calcium and supplemented with 33, 60 or 90 mM of KCl. For comparison, the effects of the calcium channel blockers nifedipine and verapamil and the activator of adenylyl cyclase forskolin were assayed on contractions induced by KCl and CaCl2. 2. Genistein (IC50: 20.2 +/- 1.0 microM, n = 11), kaempferol (IC50: 10.1 +/- 0.8 microM, n = 8) and quercetin (IC50: 13.2 +/- 0.5 microM, n = 8) relaxed the tonic contraction induced by KCl (60 mM) in a concentration-dependent way. Verapamil (IC50: 70.1 +/- 5.8 nM, n = 7), nifedipine (IC50: 8.4 +/- 0.7 nM, n = 6) and forskolin (IC50: 0.62 +/- 0.08 microM, n = 14) also relaxed the KCl-induced contraction. TPCK (3 microM) significantly antagonized the effect of quercetin, kaempferol and forskolin (P < 0.01) but did not modify the effect of genistein. 3. Spermine (1 mM) increased the effects of genistein and verapamil and antagonized the effect of quercetin but did not modify those of kaempferol and forskolin. DFMO (10 mM) did not modify the effect of quercetin but increased that of genistein and antagonized those of kaempferol and forskolin. The addition of spermine (1 mM) plus DFMO (10 mM) antagonized the effect of quercetin. Spermine counteracted the effect of DFMO on forskolin but not on genistein. 4. KCl (33, 60 or 90 mM) did not produce contraction in calcium free solution, but CaCl2 (30 microM to 10 mM) induced concentration-dependent contraction after depolarizing with KCl. The EC50 values for CaCl2 were: 0.74 +/- 0.08 (n = 12), 0.34 +/- 0.03 (n = 14) and 0.48 +/- 0.02 (n = 12) mM in a medium with 33, 60 or 90 mM of KCl, respectively. 5. Genistein (20 microM), kaempferol (10 microM), quercetin (15 microM), verapamil (70 nM), nifedipine (10 nM) and forskolin (0.5 microM) inhibited the concentration-response curve to CaCl2 in medium supplemented with 33, 60 or 90 mM of KCl. The effect of kaempferol was independent of the concentration of KCl in the incubation medium. However, the inhibitory effect of genistein on CaCl2-induced contraction was inversely related to the concentration of KCl in the medium. On the contrary, the effect of quercetin was directly related to the concentration of KCl in the medium. 6. The antagonism of verapamil, nifedipine and forskolin on CaCl2-induced contraction seems to be related to the degree of depolarization because increasing the KCl in the medium counteracted their effects. 7. Our results suggest that (1) cAMP contributes to the relaxant effects of quercetin and kaempferol on KCl (60 mM) induced tonic contraction; (2) polyamines are involved in the relaxant effects of forskolin and kaempferol on KCl-induced tonic contraction but not on CaCl2 induced contraction in the depolarized uterus, and (3) the flavonoids assayed also possess a calcium antagonist action but show a different behavior toward the calcium channel blockers and the cAMP enhancer forskolin. PMID- 9347338 TI - The cardiovascular effects of green beans (Phaseolus aureus), common rue (Ruta graveolens), and kelp (Laminaria japonica) in rats. AB - 1. The hypotensive effect of green beans, common rue and kelp was recently shown in normotensive rats in vivo. A number of mechanisms of action of these aqueous extracts was identified. The present study examined these actions at the tissue level in vitro with possible interactions of these extracts. 2. Rue showed positive chronotropic and inotropic effects on isolated right atria. Green beans and kelp alone showed negative chronotropic effects on isolated right atria but no effect on atrial tension (AT). A combination of green beans and kelp showed no additive effect on the decrease in atrial rate (AR) nor any negative inotropic responses. Combinations of rue and green beans and of rue and kelp showed responses that were either positive or negative chronotropically, were not dose dependent and were less than the sum total of their individual responses (i.e., subtractive). A combination of all three showed subtractive effects on the decrease in AR that were dose related. No change in AT was observed upon treatment with a combination of the three plant extracts in spite of the positive inotropic effect of true. 3. Rue and kelp alone relaxed KCl preconstricted rat tail artery strips probably by a direct effect of vascular smooth muscle. Green beans had no effect. The combination of rue and kelp exerted a subtractive relaxation effect. These plants therefore contained cardiovascular active substances that had a direct effect on the cardiovascular system. These substances further interacted to modify their cardiovascular effects. 4. Data explained why herbs, as in herbal medicine, should be used together therapeutically. PMID- 9347339 TI - Effect of cisapride on the isolated guinea pig gall bladder and common bile duct. AB - 1. In this study, the effect of cisapride on the isolated guinea pig gall bladder (GB) and common bile duct (CBD) motility was investigated. 2. Cisapride, up to a certain concentration, produced an increase in tone of the GB (EC50 1.35 x 10(-8) M) and the CBD (EC50 2.75 x 10(-9) M), whereas, at concentrations higher than 3 x 10(-5) M for the GB and 5 x 10(-6) M for the CBD, it produced a transient increase in tone followed by a sustained decrease in tone or in the contraction amplitude or in both. 3. The cisapride-induced increase in tone was antagonized by atropine on both the GB and the CBD. 4. Cisapride up to 2 x 10(-5) M for the GB and 3 x 10(-6) M for the CBD did not modify, whereas, at concentrations higher than 2.8 x 10(-5) M for the GB and 4 x 10(-6) M for the CBD, it antagonized, noncompetitively, the concentration-response curve to exogenously applied acetylcholine. The IC50 values for cisapride on the EC50 of acetylcholine on the GB and the CBD were 9.4 x 10(-5) M and 8.2 x 10(-6) M, respectively. 5. In conclusion, on the guinea pig GB and CBD, low concentrations of cisapride produce a stimulating effect, probably of cholinergic origin, whereas higher concentrations produce a transient stimulating effect, probably of cholinergic origin, followed by a sustained relaxing effect, which may involve both cholinergic and noncholinergic pathways. PMID- 9347340 TI - Changes in microtubular tau protein after morphine in a cultured human neuroblastoma cell line. AB - 1. Cultured human SH-SY5Y adrenergic neuroblastoma cells were used to examine the action of morphine sulfate on microtubular tau protein. 2. After 48 hr treatment morphine sulfate (200 microM) reduced tau protein in the cytoplasmic (supernatant) fraction of undifferentiated cells, and in the cytoplasmic as well as membrane (pellet) fractions of differentiated cells. 3. A 71% increase (P < 0.05) in total protein in the membrane (pellet) fraction of undifferentiated cells and a 188% increase (P < 0.01) in that of differentiated cells accompanied the decrease in tau protein. 4. A 51% reduction (P < 0.01) in the number of undifferentiated (but not differentiated) cells was seen after this drug (200 microM). PMID- 9347341 TI - Effects of ginsenosides injected intrathecally or intracerebroventricularly on antinociception induced by morphine administrated intracerebroventricularly in the mouse. AB - 1. Total saponin fraction at doses of 0.1 to 1.0 microgram, which administered IT alone did not affect the latencies of the tail-flick threshold, dose dependently attenuated inhibition of the tail-flick response induced by ICV-administered morphine (2 micrograms). 2. Total saponin fraction at doses of 1 to 20 micrograms, which administered ICV alone did not affect the latencies of the tail flick response, did not affect ICV-administered morphine-induced antinociception. 3. The duration of antagonistic action of the total saponin fraction against morphine-induced antinociception lasted for at least 6 hrs. 4. Various doses of ginsenosides Rb2, Rc, Rd and Rg1, but not Rb1 and Re, injected IT dose dependently attenuated antinociception induced by morphine administered ICV. 5. In summary, ginsenosides Rb2, Rc, Rd and Rg1 injected spinally appear to have antagonistic action against the antinociception induced by supraspinally applied morphine. On the other hand, the total ginseng fraction, at supraspinal sites, may not have an antagonistic action against the antinociception induced by morphine administered supraspinally. PMID- 9347343 TI - Septal neuronal responses related to spatial representation in monkeys. AB - Neuronal activity in the monkey septal nuclei was recorded during performance of a place-dependent go/no-go task in which reward contingencies of the objects were variable with reference to the spatial location of a monkey's cab in one of four places in an experimental room. Of 430 septal neurons recorded, 58 responded differentially to views outside the cab at the four locations of the monkey (place-differential neurons). To investigate the possibility that an ensemble of place-differential neurons represents a space by encoding different scenes (views), responses of the 58 place-differential neurons were analyzed by multidimensional scaling (MDS). The MDS transformed relationships among the four places, expressed as correlation coefficients between all possible pairs of two places based on the 58 place-differential responses, into geometrical relationships in a two-dimensional virtual space. The four places distributed at relative positions in a two-dimensional virtual space derived from the MDS were similar to those in the real experimental room. Furthermore, these correlation coefficients derived from 58 place-differential responses significantly and negatively correlated to behavioral performance in the discrimination of the four places. The results suggest that the ensemble of place-differential responses in the septal nuclei may predict behavioral performance to discriminate places and may represent a space based on the scenes viewed from different locations. PMID- 9347342 TI - Is activation of the metabotropic glutamate receptors impaired in the hippocampal CA1 area of the aged rat? AB - The effects of aging on activation of metabotropic glutamate (mGlu) receptors were studied in the CA1 field of hippocampal slices from young (3- to 4-month old) and aged (24- to 27-month-old) Sprague-Dawley rats with the use of ex vivo electrophysiological recording techniques. The depolarization of membrane potential, the increase in input resistance, and the blockade of the afterhyperpolarization induced in pyramidal cells of young rats by bath application of the mGlu receptor agonist (+/-)-trans-1-aminocyclopentate-1,3 dicarboxylic acid were not altered in aged animals. No age-related changes of the depressive effects of the mGlu receptor agonist were found on either the excitatory glutamatergic postsynaptic potential or the GABA-mediated inhibitory postsynaptic potentials induced by the stimulation of the stratum radiatum. The magnitude of synaptic plasticity involving mGlu receptor activation, although weaker, was not significantly altered in aged rats. This absence of age-related effects on activation of mGlu receptors may be important in understanding the possible origins of the alterations in neuronal plasticity which occur in brain aging. PMID- 9347344 TI - Vestibular-hippocampal interactions. AB - Since the early 1960s, researchers have speculated that the vestibular system, the sensory system concerned with the perception of balance and self-motion, contributes to spatial information processing and the development of spatial memory in the hippocampus. Anatomical studies have suggested that various parts of the thalamus are likely to transmit vestibular information to the hippocampus, perhaps via the parietal cortex; however, more direct pathways are possible. Over the last 2-3 years there have been a number of direct electrophysiological demonstrations that vestibular stimulation affects head direction cells in the anterior thalamic nuclei and place cells in the hippocampus. These studies demonstrate the importance of vestibular-hippocampal interactions for hippocampal function but also raise the possibility that the hippocampus may be important for compensation of vestibular function following peripheral or central vestibular lesions. PMID- 9347345 TI - Morphology of dentate granule cells in the human epileptogenic hippocampus. AB - Hippocampal dentate granule cells in temporal lobe epilepsy (TLE) patients with mesial sclerosis (MTLE) are reported to be hyperexcitable compared to those in patients with a mass lesion outside the hippocampus (MaTLE) (Williamson, Clin Neurosci 1994;2: 47-52). To determine if such hyperexcitability is associated with an altered morphology of these neurons, Lucifer Yellow-filled granule cells from MTLE patients were compared with those from MaTLE. The morphology of granule cells in both subject groups resembles closely that of human granule cells described previously by Golgi studies. About 40% of human granule cells have basal dendrites. Additionally their apical dendrites are much more limited in their spread in the longitudinal axis of the hippocampus contributing perhaps to a much more narrow lamellar organization than in rats. Analysis of variance computed on 21 morphometric parameters reveals a significant increase in the length of the portion of the dendrite in the inner molecular layer (IML), and a decrease in length in the outer third of the molecular layer in MTLE, compared to MaTLE. Factor analysis performed on the morphometric features of each group of neurons reveals that in the MaTLE neurons the most distinctive feature is the total dendritic length and the overall distribution of spines on them, whereas in MTLE a lengthening and elaboration of the dendrites in the IML is most distinctive. Previous observations of increased synaptic terminals containing neuropeptides, and neurotransmitter receptors in the IML taken in conjunction with an elaboration of granule cell dendrites in this region, suggest considerable synaptic reorganization within the IML of the MTLE hippocampus which may contribute to its epileptogenicity. PMID- 9347346 TI - Reduction in spine density associated with long-term potentiation in the dentate gyrus suggests a spine fusion-and-branching model of potentiation. AB - Approximately 2,700 dendritic spines in Golgi-impregnated hippocampal granule cells were quantified via image analysis 24 h after the unilateral induction of long-term potentiation in seven rats. Stereological corrections were made using a tilting disector and analytical unfolding technique. In the potentiated hemisphere the mean spine density along dendrites was reduced by approximately 20%. The relative frequency of shorter, thicker spines was increased in potentiated tissue. Physiological consequences of two morphological changes leading to a reduction in spine density (retraction or fusion of spines) were examined using a compartmental model of a simplified granule cell. The model was constructed in the NEURON modeling environment and included a realistic population of 60 dendritic spines (with dual-component synapses and active Ca(2+) dependent mechanisms). Simulations demonstrated that potentiation of postsynaptic responses was compatible with fusion (with branching) of a proportion of spines with their neighbors but was not compatible with retraction of spines. This result held over wide variations of model parameters as long as dendritic membranes were assumed to be excitable. PMID- 9347347 TI - Roles of movement and temporal factors in spatial learning. AB - Previous experiments suggested that rats can learn to discriminate between adjacent arms of an eight-arm radial maze if they have an intact hippocampal system and are allowed to move around on the maze. These requirements are consistent with the hypothesis that this discrimination involves hippocampus based spatial learning. We examined the importance of self-generated movement in this form of learning by moving rats manually ("passive movement") between two adjacent maze arms within a single training trial. Rats moved passively between arms (only one of which contained food) within trials learned to discriminate between the arms, as measured by a conditioned preference for the food arm when both arms were empty. This form of learning was impaired by lesions of fimbria fornix, but was unaffected by lesions of the lateral nucleus of the amygdala. Normal rats that were picked up and replaced on the same arm within trials and experienced their food and no food arms on different daily trials failed to learn the same discrimination. These findings suggest that self-generated movement is not required for spatial learning that may be mediated by a hippocampal system; rather, movement may simply serve to provide information from different locations about the cues in an environment. PMID- 9347348 TI - Effect of transient cerebral ischemia on gamma-aminobutyric acidA receptor alpha 1-subunit-immunoreactive interneurons in the gerbil CA1 hippocampus. AB - Following transient cerebral ischemia, pyramidal cells within area CA1 of the hippocampus exhibit delayed neuronal death. While interneurons within this sector continue to survive long-term, there is evidence that some interneurons in area CA1 are vulnerable to damage. To determine the nature of vulnerability in a neurochemically heterogeneous population of interneurons throughout area CA1, we examined the labeling of gamma-aminobutyric acid (GABA)ergic interneurons with an antibody to the GABAA receptor alpha 1-subunit 1-35 days following cerebral ischemia in the Mongolian gerbil. Unlike some other GABA interneuron markers, this antibody labels both the dendrites and soma of interneurons, allowing dendritic structure to be examined. Three to four days following ischemia, the pyramidal cells in area CA1 had degenerated, and the alpha 1-subunit-positive interneurons in all layers of area CA1 had developed severely beaded dendrites. At longer survival times (21-35 days), the alpha 1-subunit-immunolabeled dendrites of these interneurons had a fragmented appearance. In contrast, interneurons bordering str. oriens and alveus typically exhibited normal dendritic morphology. Despite the pathologic changes, there was no evidence of interneuron loss in area CA1 up to 35 days post-ischemia. Normal interneuron morphology was also observed in area CA3 and dentate gyrus, regions where neither pyramidal neurons nor granule cells, respectively, die following 5 min of cerebral ischemia. To determine if the ischemia-induced changes in interneuron morphology could be prevented, diazepam was administered 30 and 90 min following ischemia. Diazepam produces long-term neuroprotection of area CA1 pyramidal neurons. In gerbils sacrificed 35 days after ischemia, diazepam markedly attenuated the dendritic beading of the area CA1 interneurons. In addition, the dendrites did not display the fragmented labeling by the alpha 1-subunit antibody. Thus, despite their long-term survival, CA1 hippocampal interneurons in the gerbil can express severe structural abnormalities after transient cerebral ischemia coincident with pyramidal cell degeneration, and the injury to the dendrites can be prevented by the neuroprotectant diazepam. PMID- 9347349 TI - Loss of perikaryal parvalbumin immunoreactivity from surviving GABAergic neurons in the CA1 field of epileptic gerbils. AB - The Mongolian gerbil (Meriones unguiculatus) is known as a genetic model of epilepsy. Seizure behavior ranges from subtle events like arrest of motor activity and facial spasms to grand mal seizures followed by automatisms. Exploratory behavior in a stressful situation represents the most effective environment for provoking seizures in gerbils. Modifications of the inhibitory hippocampal circuits have been suggested as a cause of seizure susceptibility in the gerbil. This study presents a quantitative analysis of the hippocampal parvalbumin (PV)-immunoreactive and gamma-aminobutyric acid (GABA)-immunoreactive neurons in gerbils whose seizure sensitivity had been scored. PV is a cytosolic calcium-binding protein synthesized by a subpopulation of GABAergic neurons and thought to be responsible for the fast spiking capability of this subset of neurons. We show that the number of PV-immunoreactive neurons in the CA1 field of the gerbil hippocampus decreases in repeatedly seizing animals as compared to non seizing controls. The lowest density of PV-immunoreactive neurons was observed 1 hour after the last generalized seizure. No changes in the density of GABA immunoreactive neurons in field CA1 paralleled the obvious loss of perikaryal PV immunoreactivity. The CA1 field represents the final output region to extrahippocampal brain areas, and its recruitment or not into seizure activity is crucial for the spreading of hippocampal discharges to the adjacent neocortex. A reduction of such a calcium-buffering system in the soma and dendrites may affect the spike characteristics of PV-containing GABAergic neurons and may alter their response to glutamatergic transmission. A reduced inhibitory control of pyramidal cells may ensue, facilitating neuronal excitability as a result. PMID- 9347350 TI - Motivation-related neuronal activity in the object discrimination task in monkey septal nuclei. AB - Septal nuclei are suggested to work as an interface between the hippocampal formation, involved in higher cognitive functions, and the hypothalamus, involved in motivational behaviors such as feeding, drinking, and intracranial self stimulation. In the present study, to elucidate a role of the septal nuclei in motivational behaviors, single neuron activity was recorded from water- and food deprived monkeys during discrimination of objects associated with juice, and during ingestion of juice. Of 349 neurons recorded from two monkeys, 67 responded in the ingestion phase of the object discrimination task. Of these 67 neurons, 31 were further tested with the noncontingent liquid (juice or water) test in which liquid was provided until the animals became satiated. These 31 septal neurons were classified into two groups: type I neurons (n = 10) responded to juice ingestion with inhibition, and type II neurons (n = 21) responded with excitation. The spontaneous firing rates of the type I neurons were higher in the deprived condition and decreased as the animal became satiated by intake of liquid. Nine type II neurons responded to the sight of a white object associated with juice as well as ingestion of juice. The response magnitudes of the type II neurons to both the sight of the white object and ingestion of juice also decreased by satiation. However, spontaneous firing rates of the type II neurons did not change. These activity changes of both type I and II neurons were well correlated with changes in motivational state of the monkey estimated by the behavioral test. The results suggest that the activity of type I neurons reflects thirst or hunger drive levels, and that responses of type II neurons are related to reward perception. These type I and II neurons were located mainly in the anterior part of the septal nuclei. Results of the present study suggest, along with previous lesion and anatomical studies, that the septal nuclei exert a powerful influence on the motivational/drive systems through the projection to the hypothalamus. PMID- 9347351 TI - Role of temporal summation in age-related long-term potentiation-induction deficits. AB - Hippocampal long-term potentiation (LTP) is reduced in aged relative to young F 344 rats when peri-threshold stimulation protocols (several stimulus pulses at 100-200 Hz) are used. The present study was designed to examine the possibility that this LTP-induction deficit is caused by a reduced overlap of Schaffer collateral inputs onto CA1 pyramidal cells (input cooperativity). This reduced input cooperativity would decrease the levels of postsynaptic depolarization during LTP induction, which might account for the age-related LTP deficit. Both behavioral data (Morris Water Maze) and electrophysiological data (intracellular recordings from hippocampal slices) were collected from adult and aged F-344 rats. To counter the effects of reduced input cooperativity, stimulus intensities were adjusted to elicit baseline excitatory postsynaptic potentials (EPSPs) of equivalent amplitude in aged and young rats. Contrary to expectations, however, an age-related LTP-induction deficit was still observed. Further evaluation of the electrophysiological data revealed that temporal summation of multiple EPSPs during high-frequency stimulation was impaired in the aged rats. Thus, despite the equalization across age groups of the baseline EPSP amplitudes, the cells of aged rats were less depolarized during the LTP-inducing stimulation than were those of young rats. This reduced total depolarization was not an artifact of the higher stimulus intensity used on aged animals, nor was it caused by a failure of aged rats' CA1 afferents to follow high-frequency stimulation. The present data therefore suggest that there is a deficit in the ability of aged rats' synapses to provide the sustained depolarization necessary to active the LTP-induction cascade. PMID- 9347352 TI - Ultrastructural localization of neurotransmitter immunoreactivity in mossy cell axons and their synaptic targets in the rat dentate gyrus. AB - Electrophysiologically identified and intracellularly biocytin-labeled mossy cells in the dentate hilus of the rat were studied using electron microscopy and postembedding immunogold techniques. Ultrathin sections containing a labeled mossy cell or its axon collaterals were reacted with antisera against the excitatory neurotransmitter glutamate and against the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). From single- and double-immunolabeled preparations, we found that 1) mossy cell axon terminals made asymmetric contacts onto postsynaptic targets in the hilus and stratum moleculare of the dentate gyrus and showed immunoreactivity primarily for glutamate, but never for GABA; 2) in the hilus, glutamate-positive mossy cell axon terminals targeted GABA-positive dendritic shafts of hilar interneurons and GABA-negative dendritic spines; and 3) in the inner molecular layer, the mossy cell axon formed asymmetric synapses with dendritic spines associated with GABA-negative (presumably granule cell) dendrites. The results of this study support the view that excitatory (glutamatergic) mossy cell terminals contact GABAergic interneurons and non GABAergic neurons in the hilar region and GABA-negative granule cells in the stratum moleculare. This pattern of connectivity is consistent with the hypothesis that mossy cells provide excitatory feedback to granule cells in a dentate gyrus associational network and also activate local hilar inhibitory elements. PMID- 9347353 TI - Morphological characteristics of layer II projection neurons in the rat medial entorhinal cortex. AB - The entorhinal cortex receives inputs from a variety of neocortical regions. Neurons in layer II of the entorhinal cortex originate one component of the perforant path which conveys this information to the dentate gyrus and hippocampus. The current study extends our previous work on the electro responsive properties of layer II neurons of the medial entorhinal cortex in which we distinguished two categories of layer II neurons based on their electrophysiological attributes (Alonso and Klink [1993] J Neurophysiol 70: 128 143). Here we report on the morphological features of layer II projection neurons, as revealed by in vitro intracellular injection of biocytin. We now report that the two electrophysiologically distinct types of neurons correspond to morphologically distinct types of cells. All neurons (65% of the total cells recorded) that developed sustained, subthreshold, sinusoidal membrane potential oscillations were found to have a stellate appearance. Neurons that did not exhibit oscillatory behavior had either a pyramidal-like (32%) or a horizontal cell morphology (3%). Stellate cells had multiple, thick, primary dendrites. Their widely diverging upper dendritic domain expanded mediolaterally over a distance of around 500 microns close to the pial surface. This mediolateral extent was more than double that of the pyramidal-like cells. Dendrites of stellate cells demonstrated long dendritic appendages, and their dendritic spines had a more complex morphology than those of nonstellates. The stellate cell axons emerged from a primary dendrite and were more than double the thickness (approximately 1.4 microns) of the axons of nonstellate cells. Recurrent axonal collaterization appeared more extensive in axons arising from stellate cells than from pyramidal-like cells. PMID- 9347354 TI - Recording of mitochondrial transmembrane potential and volume in cultured rat osteoclasts by confocal laser scanning microscopy. AB - Osteoclasts are multinuclear bone-resorbing cells which contain abundant mitochondria. Morphological studies have suggested that a correlation may exist between mitochondrial concentration and bone resorption by osteoclasts. However, investigation of mitochondrial transmembrane potential (delta psi) and volume has been hampered by the difficulty in obtaining a sufficient number of osteoclasts for assessing these characteristics by flow cytometric analysis. In this study, we have used confocal laser scanning microscopy after loading the cells with Rhodamine 123 and 10-nonyl Acridine Orange to record mitochondrial delta psi and volume, respectively, in isolated rat osteoclasts cultured on bovine bone slices. Optimal staining conditions were found to be 10 micrograms ml-1 for 40 min for Rhodamine, and 1 microM for 10 min for the 10-nonyl Acridine Orange derivative. Two osteoclast populations, whose shape seemed to reflect bone resorption and migratory functions, were identified depending on their shape and on the distribution of the two dye probes. 'Round-shaped' osteoclasts had significantly higher mitochondrial delta psi and volume in the apical regions than in the basolateral portions (p < 0.00001). In contrast, mitochondrial delta psi and volume in 'irregular-shaped' osteoclasts were rather evenly distributed in both these regions (p > 0.05). Our results indicate that there is an apical polarization of mitochondria in osteoclasts corresponding to the energy demands associated with bone resorption. PMID- 9347355 TI - Localization of penultimate carbohydrate residues in zona pellucida and acrosomes by means of lectin cytochemistry and enzymatic treatments. AB - Lectins from peanuts (PNA) and soy beans (SBA) bind terminal residues of galactose (Gal) and N-acetyl-galactosamine (GalNAc) respectively. Galactose oxidase oxidizes the hydroxyl group at C-6 of terminal Gal and GalNAc blocking the binding of PNA and SBA. Binding of these lectins to sugar residues is also severely limited by the existence of terminal residues of sialic acid. In the present study, lectin cytochemistry in combination with enzymatic treatments and quantitative analysis has been applied at light and electron microscopical levels to develop a simple methodology allowing the in situ discrimination between penultimate and terminal Gal/GalNAc residues. The areas selected for the demonstration of the method included rat zona pellucida and acrosomes of rat spermatids, which contain abundant glycoproteins with terminal Gal/GalNAc residues. Zona pellucida was labelled by LFA, PNA and SBA. After galactose oxidase treatment, terminal Gal/GalNAc residues are oxidized, and reactivity to PNA/SBA is abolished. The sequential application of galactose oxidase, neuraminidase and PNA/SBA has the following effects: (i) oxidation of terminal Gal/GalNAc residues; (ii) elimination of terminal sialic acid residues rendering accessible to the lectins preterminal Gal/GalNAc residues; and (iii) binding of the lectins to the sugar residues. Acrosomes were reactive to PNA and SBA. No LFA reactivity was detected, thus indicating the absence of terminal sialic acid residues. Therefore, no labelling was observed after both galactose oxidase PNA/SBA and galactose oxidase-neuraminidase-PNA/SBA sequences. In conclusion, the combined application of galactose oxidase, neuraminidase and PNA/SBA cytochemistry is a useful technique for the demonstration of penultimate carbohydrate residues with affinity for these lectins. PMID- 9347356 TI - The use of Fluoresceincadaverine for detecting amine acceptor protein substrates accessible to active transglutaminase in living cells. AB - The use of Fluoresceincadaverine as a primary amine donor for detecting the endogenous substrates for active transglutaminase in living cells was studied. Fluoresceincadaverine was found to be suitable for labelling cells in culture as it did not induce cytotoxicity when used at 0.5 mM in culture media and diffused throughout the cell. After appropriate fixation using methanol, Fluoresceincadaverine-labelled cells were observed by direct fluorescence microscopy, allowing visualization of the substrates for active transglutaminase. Simultaneous detection of transglutaminase and of Fluoresceincadaverine incorporated into proteins strongly suggested that cytosolic transglutaminase was inactive in these living cells. However, transglutaminase co-distributed with Fluoresceincadaverine-labelled structures, which resembled a lattice. Fluoresceincadaverine-labelled proteins detected by Western blotting using an anti-Fluorescein antibody showed that, in living cells, the major transglutaminase substrate migrated at an apparent molecular weight of 220 kDa, as does fibronectin. Fibronectin was found to co-distribute with Fluoresceincadaverine-labelled lattice. This confirmed that these lattice structures were extracellular and, therefore, that transglutaminase is in an active form in this compartment. This opportunity to perform morphological and biochemical analyses in the search for transglutaminase substrates in living cells should help in determining the specific function of transglutaminases in a particular cell type as well as in universal cellular events, such as apoptosis or cell growth. PMID- 9347357 TI - Binding of antibiotics to glycoproteins of the vitelline and fertilization envelopes of cherry salmon eggs. AB - The binding of antibiotics (gentamicin, oleandomycin and chloramphenicol) to vitelline and fertilization envelopes and their extracts was investigated by immunohistochemical and immunocytochemical techniques and immunoblot analysis using mature and artificially activated eggs of the fish Oncorhynchus masou. Binding of antibiotics was detected in the vitelline and fertilization envelope outermost layers, the fertilization envelope inner surface and cortical alveolus exudates, with differences in immunoreactive intensity and deposition. The fertilization envelope outermost layer had the capacity to bind much greater amounts of the antibiotics than the vitelline envelope outermost layer. The greater capacity was caused by the deposition of cortical alveolus exudates, which were known to be responsible for functional roles of protection against bacteria, fungi and noxious materials. Treatment of the vitelline and fertilization envelopes with neuraminidase markedly reduced the binding of gentamicin and chloramphenicol but slightly increased that of oleandomycin; binding of the latter to the vitelline and fertilization envelope outermost layers was considerably reduced after treatment with alpha-fucosidase. Treatment of the two envelopes with alpha-mannosidase, beta-galactosidase or beta-D glucosaminidase did not cause any alteration in immunoreactive intensity or number of immunoreactive deposits. Immunoblot analysis of the vitelline or fertilization envelope extracts indicated that many of the antibiotic-binding substances were glycoproteins, and several major bands were bound by all three antibiotics. These results suggest that the vitelline or fertilization envelopes may have the ability to protect the egg itself, or the embryo, respectively, by trapping antibiotics, and the trapping may be related to the presence of carbohydrate moieties, such as sialyl or fucosyl residues. PMID- 9347358 TI - A histochemical study of anionic sites in the intermediate layer of rat femoral cartilage using polyethyleneimine at different pH levels. AB - Anionic sites in the intermediate layer of young rat hyaline cartilages were examined using a cationic dye, polyethyleneimine (PEI), at different pH levels. Femoral heads were resected and fixed in 2.5% glutaraldehyde and treated with 0.5% PEI at pH 7.4, pH 2.5 or pH 1.0. Some cartilage samples were first digested with chondroitinase ABC or hyaluronidase. The PEI deposits at pH 7.4 appeared to be irregular shapes. Their sizes seemed to be larger than those at pH 2.5 or pH 1.0. The PEI deposits were also found on the surface of collagen fibrils at both pH 7.4 and pH 2.5 even after the chondroitinase ABC digestion, but were not found at pH 1.0. Moreover, they disappeared after hyaluronidase digestion. Accordingly, it is suggested that PEI-positive structures varied depending on pH levels. In addition, hyaluronan may be localized near collagen fibrils, but most sulphated proteoglycans may not. PMID- 9347359 TI - Non-isotopic in situ hybridization to detect chick Sox gene mRNA in plastic embedded tissue. AB - In situ hybridization techniques have rapidly become widely used by the molecular biologist for the localization of specific nucleic acid sequences in individual cells or tissues. We describe the demonstration of Sox gene mRNA in chick tissue that has been embedded in the plastic methyl methacrylate to permit the preparation of sections for high-resolution light microscopy. Polymerization of the plastic was induced by using either N,N-dimethylaniline or N,N-3,5 tetramethylaniline. The in situ hybridization technique used was non-isotopic and used a digoxigenin-labelled probe detected with an antibody bound to alkaline phosphatase, which was then localized using X-phosphate-Nitro BT as a substrate chromogen mix. Various pretreatments of the tissue sections were investigated, including the use of proteinase K, and heat-mediated techniques using a microwave oven and a pressure cooker. The best results were produced using pressure cooking on tissue in which the plastic had been chemically polymerized with N,N-3,5 tetramethylaniline. For the demonstration of Sox 11, this combination had a critical influence on the staining results, but for Sox 21 all protocols used produced good staining. PMID- 9347360 TI - Galanin and cholecystokinin in cultured magnocellular neurons isolated from adult rat supraoptic nuclei: a correlative light and scanning electron microscopical study. AB - Cultured magnocellular neurons, isolated from adult rat supraoptic nuclei, were characterized by immunocytochemistry, using the avidin-biotin-peroxidase complex and antisera to vasopressin, oxytocin, galanin and cholecystokinin. Light microscope examination of the immunostained cultures revealed the presence of vasopressin- and oxytocin-like immunoreactivity, as well as neurons containing either galanin- or cholecystokinin-like immunoreactivity. In contrast, no significant galanin- or cholecystokinin-like immunoreactivity could be observed in freshly dispersed cells. Correlative scanning electron microscopical observations in the secondary electron imaging mode revealed that the stained neurons appeared significantly brighter than the unstained structures. Complementary observations with toad brain sections (preoptic area), immunostained for galanin, led to the same result. Considering previous results, it is suggested that the presence of galanin- and cholecystokinin-like immunoreactivity in the cultured neurons and its virtual absence in freshly dispersed cells is indicating a participation of these peptides in the regenerative processes taking place during culture. It is further concluded that the avidin-biotin-peroxidase method is suitable for correlative light and scanning electron microscopical studies of smooth surfaces and cultured cells. PMID- 9347361 TI - A rapid method for the assessment of bone architecture by confocal microscopy. AB - Conventional ways of demonstrating and analysing the components of osseous tissue have always been hampered by the difficulty of physically sectioning bone. In this study, we have used Acridine Orange staining of 100-micron-thick unembedded bone slices and then assessed the cellular and tissue architecture by confocal microscopy. The result showed the Acridine Orange, by differential staining of the cellular nucleic acids, permits ready assessment of cell shape and cell organization as well as variations in growth patterns. Our studies have provided a new and relatively easy way of assessing the morphology of bone specimens by rendering unnecessary the need for embedding, decalcification and thin sectioning of the osseous tissue. PMID- 9347362 TI - Inhaled nitric oxide: can we deliver? PMID- 9347363 TI - Optimization of patient-ventilator interactions: closed-loop technology to turn the century. PMID- 9347364 TI - Thermodilution curves obtained from the bronchial mucosa. PMID- 9347365 TI - Altering ventilation-perfusion relationships in ventilated patients with acute lung injury. PMID- 9347366 TI - Bronchial temperature as a key to the interior pulmonary capillary bed of anaesthetized dogs. AB - The terminal airways are separated from the surrounding pulmonary capillaries by a tissue layer of a few micrometers in thickness only. Therefore, it should be possible to gain information about in vivo transcapillary heat transport of the interior pulmonary vascular bed by recording the terminal bronchial temperature. For this purpose, we studied temperature-time curves in the pulmonary artery, bronchial system and the aorta of six anaesthetized dogs permanently instrumented for measuring pulmonary blood flow. Thermistors recorded temperature changes at the three locations after injection of 5 ml cold solution into the right atrium. From the observed temperature-time curves mean transit times between the three recording sites were calculated for various pulmonary blood flows (integral of delta Ttdt/integral of delta Tdt) (range 1.1-3.5 1/ min). We found that the temperature-time curves of the bronchial system resemble typical "dilution" curves and are interspaced between those in the pulmonary artery and those in the aorta. Regardless of pulmonary blood flow, mean transit times from the pulmonary artery to the distal bronchial system and from there to the aorta were about equal. We conclude that transcapillary heat transfer generates bronchial temperature-time curves which permit an estimation of the relation of precapillary to postcapillary mean transit times in the interior of the lung. PMID- 9347367 TI - Sequential use of noninvasive pressure support ventilation for acute exacerbations of COPD. AB - OBJECTIVES: To compare the efficacy of noninvasive pressure support ventilation (NIPSV) in acute decompensation in chronic obstructive pulmonary disease (COPD) by means of a bi-level positive airway pressure support system (BiPAP) in a sequential mode with medical therapy alone; to assess the short-term physiologic effects of the device on gas exchange; and to compare patients successfully ventilated with NIPSV with those in whom NIPSV failed. DESIGN: A prospective case series with historically matched control study. SETTING: A general intensive care unit (ICU) of a university hospital. PATIENTS: We evaluated the efficacy of administration of NIPSV in 42 COPD patients and compared this with standard treatment in 42 matched historical control COPD patients. INTERVENTIONS: NIPSV was performed in a sequential mode, i.e., BiPAP in the spontaneous mode was used for at least 30 min every 3 h. Between periods of ventilation, patients could be systematically returned to BiPAP when the arterial oxygen saturation was < 0.85 or when the respiratory rate was > 30 breaths/min. MEASUREMENTS AND RESULTS: Success rate, mortality, duration of ventilatory assistance, and length of ICU stay were recorded. Eleven of the 42 patients (26%) in the NIPSV group needed tracheal intubation compared with 30 of the 42 control patients (71%). The 31 patients in whom NIPSV was successful were ventilated for a mean of 6 +/- 3 days. In-hospital mortality was not significantly different in the treated versus the control group, but the duration of ventilatory assistance (7 +/- 4 days vs 15 +/- 10 days, p < 0.01) and the length of ICU stay (9 +/- 4 days vs 21 +/- 12, p < 0.01) were both shortened by NIPSV. BiPAP was effective in correcting gas exchange abnormalities. The pH values, measured after 45 min of BiPAP with optimal settings, in the success (7.38 +/- 0.04) and failure (7.28 +/- 0.04) patients were significantly different (p < 0.05). CONCLUSIONS: NIPSV, performed with a sequential mode, may be used in the management of patients with acute exacerbations of COPD. PMID- 9347368 TI - Postcardiac surgery low cardiac output syndrome: dopexamine or dopamine? AB - OBJECTIVE: To compare the efficacy and safety of dopexamine with dopamine in the treatment of low cardiac output syndrome after cardiac surgery. DESIGN: This was a multicentre, double-blind, randomised, parallel-group study conducted in intensive care units at centres in Holland and Belgium. Patients were randomised to receive dopexamine (up to 2.0 micrograms/ kg per min) or dopamine (up to 6.0 micrograms/kg per min) for 6 h after low cardiac output syndrome was confirmed. RESULTS: 70 patients were enrolled (35/group) and there was no significant differences in the operative procedures or haemodynamics at entry into the study. Clinical efficacy, defined as a cardiac index > 2.5 l/min per m2 with urine production > 0.5 ml/kg per h and stable haemodynamics for two consecutive readings 1 h apart, was achieved by 90 and 87% of patients in the dopexamine and dopamine groups, respectively. However, more patients maintained clinical efficacy over the 6-h period in the dopexamine group, which was statistically significant at 1-2 h and approached significance at all other time points. Safety was assessed by comparing the adverse events and concomitant medication. Fewer patients on dopexamine had cardiac events compared with dopamine-treated patients (25 vs 38 events), although there was no difference in the pattern of rhythm disturbance. Fewer patients in the dopexamine group required concomitant vasodilating drugs (18 vs 30). CONCLUSION: Taking the proportion of patients achieving clinical efficacy, the time to achieve it and the maintenance of it along with the adverse event profile, dopexamine was shown to be an effective and safe drug to use in the management of low cardiac output syndrome after coronary artery bypass graft surgery and may be superior to dopamine. PMID- 9347369 TI - Propofol in patients needing long-term sedation in intensive care: an assessment of the development of tolerance. A pilot study. AB - OBJECTIVE: To evaluate the possible development of tolerance in patients sedated with propofol in intensive care for more than 5 days. DESIGN: An open within patient non-comparative pilot study. SETTING: The intensive care unit of a hospital in the UK. PATIENTS: Twenty-three patients who were expected to need sedation with propofol for more than 5 days were included; of these, 11 were sedated continuously for between 5 and 10 days. INTERVENTIONS: Propofol was infused continuously at about 0.5-4 mg/kg per h to provide sedation at level 3 on the Ramsay scale (drowsy or asleep, responds easily to commands). Alfentanil was infused at a rate of about 0.5 microgram/kg per min. MEASUREMENTS AND RESULTS: Patients with a statistically significant increase in both infusion rate and blood concentration were considered to show tolerance, those that required an increasing infusion rate and constant (or decreasing) blood concentration were said to show increased clearance, and those that required constant (or decreasing) infusion rates and blood concentrations were classed as showing no tolerance. Of the 11 patients who were sedated for more than 5 days, three showed tolerance and three showed increased clearance while five showed no tolerance. The desired sedation level was achieved for the majority of the time for patients sedated between 5 and 10 days (mean 73-90%). There was some suggestion of a relationship between improving health (decreasing APACHE II scores) and the need for increased infusions of propofol. CONCLUSIONS: Although there were insufficient data to reach firm conclusions, the possibility of some relationship between the increasing need for propofol and improving condition in patients needing longer term intensive care cannot be ruled out. PMID- 9347370 TI - Patient-initiated, pressure-regulated, volume-controlled ventilation compared with intermittent mandatory ventilation in neonates: a prospective, randomised study. AB - OBJECTIVE: To compare the effects of patient-initiated, pressure-regulated, volume-controlled ventilation (PRVC) with pressure-preset intermittent mandatory ventilation (IMV) in neonates with respiratory failure. DESIGN: Randomised, prospective study. SETTING: Intensive care unit (14 beds) in a 300-bed paediatric teaching hospital. PATIENTS: 60 neonates with respiratory distress syndrome (RDS) or congenital pneumonia, weighing < 2500 g and requiring mechanical ventilation. INTERVENTIONS: Ventilatory support until extubation via either IMV (n = 30) or PRVC (n = 27). In PRVC, the tidal volume (VT) was preset and pressure-controlled breaths delivered with peak inspiratory pressure values adapted to achieve the preset VT. MEASUREMENTS AND RESULTS: Main outcome measures were duration of ventilation and incidence of bronchopulmonary dysplasia (BPD). Pulmonary air leaks and intraventricular haemorrhage (IVH) were considered major adverse effects. Demographic data, ventilation parameters and arterial/alveolar oxygen tension ratio were similar at randomisation. Duration of ventilation and incidence of BPD were not decreased by the use of PRVC. Air leaks occurred in 3 neonates in the PRVC group and in 7 babies treated with IMV (NS). The incidence of IVH grade > II was lower in babies treated with PRVC (p < 0.05). In a subgroup of neonates weighing < 1000 g, the duration of ventilation and incidence of hypotension were reduced in the PRVC group (p < 0.05). CONCLUSION: Patient initiated, pressure-regulated, volume-controlled ventilation can be safely used in neonates and may contribute to a lower incidence of complications. PMID- 9347371 TI - Changes in left ventricular function in shocked newborns. AB - OBJECTIVE: To assess whether the change in cardiac output after volume replacement is due to elevation of stroke volume or heart rate and to determine the effect of mechanical ventilation on the hemodynamic situation. DESIGN: Prospective study. SETTING: A ten-bed neonatal intensive care unit (level III) at a university hospital. PATIENTS: 15 consecutive newborns with blood pressure below the 10th percentile related to age and weight. INTERVENTIONS: Volume replacement with Ringer's lactate 20 ml/kg body weight. MEASUREMENTS AND RESULTS: Before and after volume replacement, arterial pressure recordings, blood gas analysis, and an echocardiographic study were carried out. Left ventricular and aortic diameters were measured by the two-dimensional M-mode technique and velocity time integral of aortic flow by the pulsed color Doppler technique. From these data, stroke volume and cardiac output were calculated. Cardiac output (703 +/- 204 vs 826 +/- 166 ml/ min, p < 0.005) and cardiac index (267 +/- 69 vs 302 +/- 55 ml/min per kg body weight, p < 0.01) changed significantly due to an appreciable elevation in stroke volume (5.2 +/- 1.7 vs 5.8 +/- 1.7 ml, p < 0.05), whereas heart rate was unaltered (140 +/- 12 vs 142 +/- 20 beats/min; NS). The change in blood pressure (32 +/- 5 vs 38 +/- 8 mm Hg, p < 0.01) was also significant. Cardiac index before and after volume replacement showed a significant inverse correlation with the severity of respiratory disease expressed as alveolar-arterial oxygen difference (A-aDO2) (A-aDO2 vs cardiac index before volume replacement: r = -0.77, p < 0.001; after volume replacement: r = -0.73, p < 0.005) or oxygenation index (oxygenation index vs cardiac index before volume replacement: r = -0.73, p < 0.005; after volume replacement: r = 0.73, p < 0.005). Changes in left ventricular diastolic diameter, left ventricular systolic diameter, and fractional shortening were not significant. CONCLUSIONS: These results indicate that the major regulator of left ventricular output in newborns with hypovolemic or cardiogenic shock is stroke volume and not heart rate and that cardiac output depends on the severity of the respiratory disease. PMID- 9347372 TI - Clinical validation of cardiac output measurements using femoral artery thermodilution with direct Fick in ventilated children and infants. AB - OBJECTIVE: To validate clinically cardiac output (CO) measurements using femoral artery thermodilution in ventilated children and infants by comparison with CO estimated from the Fick equation via a metabolic monitor. DESIGN: Prospective, comparison study. SETTING: Paediatric intensive care unit of a university hospital. PATIENTS: 24 ventilated infants and children, aged 0.3 to 175 months (median age 19 months). INTERVENTIONS: Oxygen consumption measurements were made and averaged over a 5-min period, at the end of which arterial and mixed venous blood samples were taken and oxygen saturations measured by co-oximetry, with CO being calculated using the Fick equation. Over this 5-min period, five sets of femoral arterial thermodilution (FATD) measurements were made and averaged. One comparison of CO values was made per patient. RESULTS: Mean Fick CO was 2.55 l/min (range 0.24 to 8.71 l/min) and mean FATD CO was 2.51 l/min (range 0.28-7.96 l/min). The mean bias was 0.03 l/min (95% confidence interval -0.07 to 0.14 l/min), with limits of agreement of -0.45 to 0.52 l/min. When indexed to body surface area, the mean Fick cardiac index became 3.51 l/min per m2 (1.52-6.98 l/min per m2) and mean FATD 3.49 l/min per m2 (1.74-6.84 l/min per m2). The mean bias was 0.02 l/min per m2 (95% confidence interval -0.11 to 0.15 l/min per m2) with limits of agreement of-0.57 to 0.61 l/min per m2. The mean FATD coefficient of variation was 5.8% (SEM 0.5%). CONCLUSIONS: FATD compares favourably with Fick derived CO estimates in infants and children and may represent an advance in haemodynamic monitoring of critically ill children. PMID- 9347373 TI - Intratracheal furosemide in infants after cardiac surgery: its effects on lung mechanics and urinary output, and its levels in plasma and tracheal aspirate. AB - OBJECTIVE: Recent studies have suggested direct pulmonary effects of furosemide in asthmatics and infants with bronchopulmonary dysplasia. We tested the hypothesis that intratracheally administered furosemide also increases respiratory compliance in children after cardiac surgery, and investigated whether furosemide has a topical and/or systemic action. STUDY DESIGN: Prospective study with intra-individual control. In twelve infants and toddlers (age: 10 +/- 8 months, weight: 6.9 +/- 3 kg) mechanically ventilated for compromised lung mechanics after cardiac surgery, 0.5 mg/kg furosemide was intratracheally administered to the lungs. Lung mechanics were serially assessed using a computerised system (Sensormedics 2600) during a 2 h control and 2 h intervention period. Urine output was measured by an indwelling bladder catheter and levels of furosemide were determined in blood and tracheal aspirates. RESULTS: Static compliance improved within 30 min in all patients, reached a maximum of 44 (20-85)% above baseline and remained improved throughout the study (p < 0.05). An immediate, short and significant diuretic effect of intratracheally applied furosemide was observed. Furosemide levels 1 h after intervention were 795 ng/ml in the blood and 431 micrograms/ml (i.e. 1000-fold higher) in the tracheal aspirate. Changes in compliance were correlated only to urine output values over the 2 h (r = 0.82, p = 0.044, n = 9) after furosemide administration. CONCLUSION: We conclude that intratracheally applied furosemide improves static compliance in infants and toddlers with compromised lung mechanics after cardiac surgery. We demonstrated that furosemide is absorbed from the lung and has a systemic effect within 15 min after its intratracheal instillation. PMID- 9347374 TI - Design flaw can convert commercially available continuous syringe pumps to intermittent bolus injectors. AB - OBJECTIVE: To investigate if unexpected behaviour of neonatal and paediatric patients connected to syringe pumps could be explained by transient elevation of these devices. DESIGN: Five different commercially available syringe-pumps were set at an infusion rate of 1 ml/h and then subjected to a vertical displacement manoeuvre (height 1 m). The actual delivered infusion volumes in association with the displacement manoeuvre were measured by a high precision weight scale connected to a computer. SETTING: A medical technology laboratory in a university hospital. MEASUREMENTS AND RESULTS: Elevation of the devices resulted in a rapid bolus injection of 0.19-2.28 ml. Returning the devices to their original positions resulted in an aspiration into the system of 0.06-0.34 ml. The times both for bolus injection and for aspiration into the system were less than 1 min in all cases. The updown manoeuvre was followed by a period with zero infusion ranging from 8 to 105 min. CONCLUSIONS: Design flaws in the construction of syringe pumps can expose patients to substantial danger following vertical displacement if potent drugs are being infused. If potent drugs are infused, care should be taken not to change the vertical position of the syringe pump even for short periods of time. Before buying new equipment, the authors recommend that the delivery characteristics of these devices should not only be tested during ordinary bench testing but should also include the reaction to a vertical displacement manoeuvre. PMID- 9347375 TI - Surgical management of Candida suppurative thrombophlebitis of superior vena cava after central venous catheterization. AB - Septic deep venous thrombosis is a major complication associated with central venous catheterization in intensive care units. The most common causative organisms are Staphylococcus aureus, gram-negative bacilli and Candida species. The incidence of Candida infections is increasing, especially in intensive care patients receiving total parenteral nutrition and long-term broad-spectrum antibiotics. Although intravascular catheter-induced septic thrombophlebitis is quite common, superior vena cava obstruction is a rare complication. However, few data exist concerning the best strategy for managing septic thrombophlebitis, especially when medical therapy fails. We report successful surgical management of Candida albicans suppurative thrombosis of the superior vena cava in a young patient. PMID- 9347377 TI - Intubation and mechanical ventilation avoided by using almitrine bismesylate in an acute hypoxemic pneumonia. PMID- 9347376 TI - Relationship between pupil size and acetylcholinesterase activity in patients exposed to sarin vapor. AB - OBJECTIVE: To elucidate the effect of sarin vapor on pupil size and erythrocyte acetylcholinesterase activity (AchE). DESIGN: Retrospective observational survey. SETTING: Emergency department of an urban teaching hospital. PATIENTS: 80 patients who were exposed to sarin in a terrorist attack in Tokyo subways. MEASUREMENTS AND RESULTS: Pupil size and AchE activity on the day of exposure were measured. Among the 80 patients, the pupils were miotic (< 3 mm) in 50 patients (62.5%), while AchE activity was below the normal range (< 1.2 U) in 34 patients (42.5%). AchE was significantly lower in the miotic group than in the group with normal pupils (1.0 +/- 0.5 U vs 1.5 +/- 0.3 U, p < 0.01). In the miotic group, AchE activity was lower than normal in 32 patients (64.0%) but was decreased in only 2 patients in the normal pupil group (6.7%) (p < 0.01). CONCLUSIONS: Miosis is a more sensitive index of exposure to sarin vapor than erythrocyte AchE. Systemic poisoning is apparently less likely to develop if the patient's pupil size is normal on arrival at the hospital. PMID- 9347378 TI - Moclobemide and imipramine in chronic depression (dysthymia): an international double-blind, placebo-controlled trial. International Collaborative Study Group. AB - An international, multicenter, placebo-controlled study was undertaken to determine the safety and antidepressant efficacy of moclobemide, a new reversible inhibitor of monoamine oxidase A, and imipramine in the treatment of dysthymia (DSM-III-R). A total of 315 patients were enrolled and randomly assigned to an 8 week treatment in one of three groups (moclobemide, imipramine and placebo). Patients were male or female outpatients aged between 18 and 65 years meeting DSM III-R criteria for dysthymia, primary type, with late or early onset. Of the patients in each group 85% completed the 8-week treatment period. The percentage of patients who no longer fulfilled DSM-III-R symptom criteria at treatment endpoint was significantly higher in the moclobemide (60%) and imipramine (49%) treatment groups than in the placebo group (22%). Differences to placebo were also statistically significant both for moclobemide and for imipramine on the other efficacy variables (i.e. Hamilton Rating Scale for Depression, final overall efficacy assessment, Clinical Global Impression and symptom check list self-rating). A significant superiority of moclobemide and imipramine over placebo was found in pure dysthymia and in double-depression, as well as in early and late onset subgroups. In early onset cases, moclobemide was significantly more effective than was imipramine on the Hamilton Rating Scale for Depression. Anticholinergic symptoms and sleepiness were significantly more frequent side effects on imipramine than on moclobemide or on placebo, and the investigators' final overall assessment of tolerability significantly favoured moclobemide over imipramine. This study demonstrates the efficacy of high dose moclobemide (mean dose 675 mg/day) and high dose imipramine (220 mg/day) against placebo in the treatment of dysthymia. Moclobemide was better tolerated than was imipramine. PMID- 9347379 TI - Long-term treatment of psychotic (delusional) depression with fluvoxamine: an open pilot study. AB - The aim of this open pilot study was to evaluate the efficacy of fluvoxamine in the continuation as well as in the maintenance therapy of delusional depression. Thirty patients with recurrent, unipolar depression (DSM-IV criteria) were selected who had at least one depressive episode during the 18 months preceding the delusional depressive index episode and were treated with fluvoxamine 300 mg/day. Twenty-five of them had a sustained response to this short-term treatment and agreed to enter into the 30-month follow up study. All participants completed the follow up period. No relapse was observed during the 6 months of continuation therapy. During the further 24 months of maintenance therapy, 80% of the patients remained well, whereas 20% (five out of 25) had a single recurrence. Based on these observations, fluvoxamine might be a promising drug for long-term therapy of delusional depression. Further controlled studies are required to confirm this finding. PMID- 9347381 TI - The influence of lithium on fluvoxamine therapeutic efficacy and pharmacokinetics in depressed patients on combined fluvoxamine-lithium therapy. AB - The influence of lithium on fluvoxamine therapeutic efficacy, plasma concentrations and pharmacokinetics was studied in 12 depressed inpatients. Six patients were on fluvoxamine monotherapy and six were on combined fluvoxamine lithium therapy. The treatment response was determined using 17-item Hamilton Rating Scale for Depression. Blood samples were collected during 48 h after a single dose administration of 100 mg fluvoxamine, and five times at steady state after repeated doses of 100 mg fluvoxamine per day. The evaluation of 17-item Hamilton Rating Scale for Depression Scores showed a significant clinical improvement 2 and 4 weeks after the beginning of the therapy in both groups (p < 0.01). However, 2 weeks after the administration of the drug(s) had started, significant differences (p < 0.05) in efficacy between the two treatments in favour of the fluvoxamine-lithium combination were found. Plasma concentrations of fluvoxamine were measured by high-performance liquid chromatography. The comparison of the measured concentrations of fluvoxamine showed a similar course of the plasma concentration-time curves in both groups of patients. Pharmacokinetic parameters of fluvoxamine did not show any significant difference on the comparison between the groups. According to the results from this study, it is evident that lithium does not affect plasma concentrations and pharmacokinetics of fluvoxamine in depressed patients on concomitant treatment with these two drugs. However, the effect achieved with the combination was better. PMID- 9347380 TI - Benefits of trazodone and mianserin for patients with late-life chronic schizophrenia and tardive dyskinesia: an add-on, double-blind, placebo-controlled study. AB - The objective of the present study was to evaluate the efficacy of mianserin and trazodone as antidepressants with serotonin 2 antagonist properties on negative symptoms and tardive dyskinesia in elderly patients with chronic schizophrenia. In this double-blind, placebo-controlled study the dose of each drug was increased gradually, from 20 mg/day mianserin to 60 mg/day, and from 50 mg/day trazodone to 200 mg/day. Symptoms were assessed using the Brief Psychiatric Rating Scale, the Scale for Assessment of Negative Symptoms and the Abnormal Involuntary Movement Scale every week for 5 weeks. A total of 38 patients (23 men and 15 women) completed the trial. Mianserin (n = 13) and trazodone (n = 12) did not alter the Brief Psychiatric Rating Scale positive symptom factor over the 5 weeks. In the mianserin group, the Scale for Assessment of Negative Symptoms total score decreased significantly after 5 weeks. Scores of 'affective flattening and blunting' and 'alogia' scores on the Scale for assessment of Negative Symptoms decreased significantly in both treatment groups. In the trazodone group, the decrease in the Abnormal Involuntary Movement Scale total score was statistically significant at weeks 2 and 3. Results indicate that serotonergic antidepressants, when used in conjunction with neuroleptics, are safe and effective for treating negative symptoms in elderly patients with chronic schizophrenia. Results also indicated a possible beneficial effect of trazodone in treating tardive dyskinesia. PMID- 9347382 TI - Trihexyphenidyl treatment of clozapine-induced hypersalivation. AB - The objective of this study was to investigate the efficacy of the anticholinergic agent trihexyphenidyl in the treatment of clozapine-induced hypersalivation. Fourteen chronic schizophrenic patients who exhibited nocturnal hypersalivation during clozapine treatment were coadministered trihexyphenidyl (5 15 mg/day, at bedtime) for 15 days. Salivation was assessed by a single-item 5 point scale. A reduction of 44% in the reported nocturnal hypersalivation was observed after trihexyphenidyl treatment. These results indicate that at least some chronic schizophrenic patients with clozapine-induced nocturnal hypersalivation may benefit from anticholinergic treatment. PMID- 9347383 TI - Incidence of antidepressant drug use in older adults and association with chronic diseases: the Rotterdam Study. AB - A follow-up study was conducted among men and women aged 55 years and over living in the community in order to estimate the incidence of initiation of antidepressant drug use and the association with chronic diseases. The study population consisted of 7,812 individuals. Overall, the incidence density for starting therapy with an antidepressant drug was 13.5 per 1000 person-years. The cumulative incidences after 1, 2 and 3 years were 1.3, 2.7 and 4.0%, respectively. The incidence in women was almost twice that in men and slightly higher in participants older than 70 years than in those younger than 70 years. The majority of the antidepressants prescribed were tricyclic antidepressants (65%), followed by selective serotonin reuptake inhibitors (23%) and other (12%) antidepressants. Only a minority (23%) received a dose considered effective for the indication of depression. Selective serotonin reuptake inhibitors were more often prescribed in an adequate dosage (68%) than were tricyclic antidepressants (12%) and other antidepressants (8%). Of the chronic diseases studied, only osteoarthritis and a history of stroke were predictors of initiation of antidepressant drug use after adjustment for age, sex and medical consumption. Hypertension, history of myocardial infarction, diabetes mellitus, rheumatoid arthritis, glaucoma, cognitive impairment and Parkinson's disease were not associated with future antidepressant drug use. No relevant differences were observed with respect to the choice of type of antidepressant drug among patients with chronic diseases. The present study indicates that each year antidepressant drug therapy is initiated in approximately 1.3% of the elderly. In general, the presence of chronic somatic diseases was not predictive of initiation of antidepressant drugs. Tricyclic antidepressants in this age group and in patients with certain chronic diseases may not be the optimal choice given their side effects profile and drug-drug and drug-disease interactions. The predominance of these agents in the present study calls for further attention. PMID- 9347384 TI - Carbamazepine toxicity induced by clarithromycin coadministration in psychiatric patients. AB - Seven psychiatric inpatients receiving carbamazepine 600 mg/day were coadministered clarithromycin 400 mg/day for 5 days to treat atypical pneumonia. Blood samples were taken after clarithromycin coadministration and at 1 and 4 weeks after its discontinuation. Plasma concentrations of carbamazepine and carbamazepine-10,11-epoxide were measured using high-performance liquid chromatography. During clarithromycin coadministration, four out of the seven patients developed moderate-to-severe toxic symptoms of carbamazepine, such as drowsiness, dizziness, and ataxia, which resolved within 5 days after clarithromycin discontinuation. In these four patients, plasma carbamazepine concentrations after clarithromycin coadministration were approximately twice as high as those after its discontinuation. In the seven patients, the mean plasma concentration of carbamazepine, but not of carbamazepine-10,11-epoxide, after clarithromycin coadministration was significantly (p < 0.01) higher than those at 1 and 4 weeks after its discontinuation. The present report suggests that clarithromycin coadministration induces increased plasma carbamazepine concentrations, which may result in carbamazepine toxicity. Therefore, care should be given to prescribing clarithromycin for patients receiving carbamazepine. PMID- 9347385 TI - An open trial of moclobemide in the treatment of post-traumatic stress disorder. AB - Traditional monoamine oxidase inhibitors have shown efficacy in the treatment of post-traumatic stress disorder, but their use is limited by some serious drug and food interactions. Moclobemide, which is a reversible inhibitor of monoamine oxidase-A, is relatively free of these limitations and is therefore potentially useful in the treatment of post-traumatic stress disorder. Twenty patients who met Diagnostic and Statistical Manual of Mental Disorders, Third Edition Revised (DSM-III-R) criteria for post-traumatic stress disorder were entered into a 12 week open study with moclobemide. Assessments were completed every 4 weeks. Eleven participants no longer met DSM-III-R criteria for post-traumatic stress disorder by week 12. The severity of post-traumatic stress disorder reduced by 2.09 SD (95% confidence interval 1.49-2.69; p < 0.001) and functional impairment improved by 1.08 SD (95% confidence interval 0.46-1.69; p < 0.01). Adverse events were minimal. Controlled, double-blind studies should be considered to confirm these findings. PMID- 9347386 TI - Fast-onset antidepressants. AB - Research on accelerating the response to antidepressants has focused attention on the possibility of fast-acting antidepressants. A major advance has been the development of methodology for identifying rapid-response antidepressants. Such antidepressants are likely to be widely used in the future and should bring considerable relief to sufferers and to the community. How rapid the action of these drugs will be is not yet clear. Careful methodology and well conducted studies are needed to clarify the issues involved. PMID- 9347387 TI - The mesolimbic dopamine system as a target for rapid antidepressant action. AB - Chronic treatment with antidepressant drugs produces a variety of changes in dopaminergic neurotransmission, most notably a sensitization of behavioural responses to agonists acting at dopamine D2/D3 receptors within the nucleus accumbens. Evidence from animal models of depression (the forced swim test and the chronic mild stress procedure) indicates that these effects are crucial for the therapeutic effect of antidepressants in these models. Antidepressant-like effects in animal models are also seen with drugs that act directly on the dopaminergic system. Because of its prolonged time-course, the chronic mild stress procedure can be used to examine onset latencies. Some dopamine-active drugs (e.g. the catechol-O-methyltransferase inhibitor tolcapone; D2/D3 agonists administered intermittently) are active in this procedure but have a time-course comparable to that of conventional antidepressants. Other dopamine-active drugs may have a more rapid onset; the evidence to date suggests this possibility for the D2/D3 agonist pramipexole and the preferential presynaptic antagonist amisulpride. In clinical studies, rapid-onset latencies have been claimed for the D2/D3 agonist roxindole, the preferential presynaptic antagonist sulpiride and the relatively selective dopamine-uptake inhibitor amineptine. The mechanisms that might give rise to a rapid onset of dopamine-mediated antidepressant effects are discussed. PMID- 9347388 TI - Pharmacology of amineptine, an antidepressant agent acting on the dopaminergic system: a review. AB - Amineptine is a tricyclic antidepressant agent with a unique capacity to reduce dopamine uptake selectively in vitro: this effect is also obtained in vivo. In vivo, amineptine increases striatal homovanillic acid without affecting the levels of other metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and 3 methoxytyramine. However, relatively high doses of amineptine preferentially lower the extracellular DOPAC level, assessed by pulse voltammetry, in the nucleus accumbens but not in the striatum. Microdialysis techniques confirm an increase in extracellular dopamine in various brain areas (striatum, nucleus accumbens and frontal cortex) and an increase in extracellular noradrenaline in the frontal cortex and dorsal hippocampus. Chronic treatment with amineptine induces downregulation of dopamine D2, beta- and alpha 2-adrenergic receptors. Amineptine enters the brain and its pharmacological effects are probably induced by the unchanged drug, rather than its two main metabolites. PMID- 9347389 TI - Early onset of therapeutic action in depression and greater efficacy of antidepressant treatments: are they related? AB - Until recently, several weeks of treatment were required to obtain a clinically significant antidepressant response using pharmacotherapy. Now treatment strategies have been developed that appear to produce an early onset of action in major depression. In treatment-resistant depression, there are several agents that can be used to potentiate the therapeutic effect of the initial antidepressant drug. The question of whether such a rapid onset is related to greater efficacy of antidepressant treatments was examined on the basis of the putative mechanisms of action of these treatments, and on the clinical evidence available so far. Some approaches appear to have both a more rapid onset of action and greater efficacy, such as electroconvulsive shock treatment, venlafaxine and pindolol addition, whereas the addition of lithium does not produce a more rapid onset of action despite being effective in treatment resistant depression. In contrast, amineptine exerts an early psychostimulant effect but is apparently as effective as other antidepressant drugs. We conclude that a treatment strategy producing a rapid onset often, but not invariably, has greater efficacy than a treatment producing a slower onset. These preclinical and clinical observations may help to devise rapid treatments for major depression that will be effective in a greater proportion of patients than at present. PMID- 9347390 TI - Early onset of action of amineptine. AB - A priority in the treatment of depression is to obtain rapid improvement at an early stage. Since depressed patients, who are often convinced that nothing can be done for them, may well have difficulty in adhering to the therapeutic management plan, they can be both uncooperative and neglectful of treatment measures. The rapid correction of this often resigned apathy is an essential aspect of treatment. According to a variety of clinical criteria, amineptine often achieves rapid improvement, particularly on measures of psychomotor retardation. Initially, antidepressant medication is an essential measure in the relief of depressive symptoms, although subsequently, it may also become a complement to psychotherapeutic support. Amineptine has been shown to act directly on the dopaminergic pathway, unlike other antidepressants, which act on this system only via their effects on the serotonergic or noradrenergic systems. PMID- 9347392 TI - Efficacy of amineptine in the prevention of relapse in unipolar depression. AB - The clinical properties of amineptine, a mainly dopaminergic antidepressant, were assessed in a double-blind controlled study involving patients fulfilling Diagnostic and Statistical Manual of Mental Disorders (DSM-III) criteria for unipolar depression. The aim was to determine how relapses could be prevented in this frequently recessing disorder. The study was a two-phase, 12-month, multicentre trial of patients suffering from major depression or dysthymia, diagnosed using DSM-III criteria and evaluated on the Montgomery-Asberg Depression Rating Scale and the Mood, Anxiety, Retardation, Danger scale. Phase I was an open-label 3-month period, with the patients being given 200 mg amineptine per day. The second, 9-month period was a placebo-controlled prophylactic phase. A total of 458 patients were initially included in the study. Of the 376 who completed phase I, 303 (66%) were responders; 284 entered the prophylactic study, randomly assigned to two groups. Of the 134 patients in the placebo group who completed phase II, 25 (18.7%) suffered a relapse, compared with nine out of the 136 (6.6%) in the amineptine group. After resolution of an acute episode of major depression or dysthymia, long-term antidepressant therapy with amineptine significantly reduced the relapse rate. PMID- 9347391 TI - The efficacy and acceptability of amineptine versus fluoxetine in major depression. AB - The temporal dimension, particularly anticipation, appears to be a very important component in the understanding of depressed patients. In a 90-day multicentre study, the efficacy and acceptability of amineptine and fluoxetine were compared in 169 patients with major depression. Comparison of the two antidepressants was based on double-blind methods, after random allocation of the treatments between two parallel groups. The two drugs did not differ over the whole course of the study, but the improvement in scores on day 4 was globally more marked in the amineptine than the fluoxetine group. Intragroup analysis showed that amineptine was significantly superior to fluoxetine on the retardation pole of the mood, anxiety, retardation, danger scale. The positive effect of amineptine on anticipation may enable the depressed patient to make plans for the future. Anticipation may be a key dimension to be more precisely explored in specific psychopharmacological protocols with antidepressants. PMID- 9347393 TI - Confusion in the courtroom. How judges have assessed the criminal responsibility of individuals with multiple personality disorder. PMID- 9347394 TI - Impact of coercion on treatment outcome. PMID- 9347395 TI - Judging the suitability for release of patients from a maximum security hospital by hospital and community staff. PMID- 9347396 TI - Youthful prostitution and child sexual trauma. AB - This paper has examined research that attempts to explain entry to prostitution in terms of the family experiences of young prostitutes. Though there is some evidence of rape, incest, and other kinds of sexual trauma in these backgrounds, this evidence is inconsistent and contradictory. A more plausible approach to the question is based on general control theories. Any traumas or conflicts that unattach children and youth from their families make youngsters highly vulnerable to delinquency. In the case of adolescent females, breach of family attachments appears to heighten the risk of early sexual involvements that, in the context of gender differences in sexual development, expose them to partners significantly older than themselves, and in significantly larger numbers than would otherwise be the case. These factors help explain the role of dysfunctional backgrounds in entry to prostitution without presupposing a role for unobservable traumas and psychiatric disturbances. They likewise recognize a role for the interaction between social control factors and the normal process of sexual development. PMID- 9347397 TI - Self-incineration. A review of the psychopathology of setting oneself afire. PMID- 9347398 TI - Recognizing and handling problems of incompetent deaf defendants charged with serious offenses. PMID- 9347399 TI - Social factors and compulsory detention of psychiatric patients in the U.K. The role of the approved social worker in the 1983 Mental Health Act. PMID- 9347400 TI - Genetic susceptibility to visceral obesity and related clinical implications. AB - This paper reviews evidence supporting the notion that genetic factors may have an influence on the determination of body fat distribution, particularly emphasizing the genetic susceptibility of visceral adipose tissue (AT) accumulation. The potential contribution of genetic susceptibility to the development of metabolic alterations in visceral obese individuals will also be reviewed. The contribution of genetic factors to the variation in body fat distribution is supported by studies in which racial differences in body fat distribution were reported. These ethnic differences suggest that body fat distribution may be influenced by some components of the genetic background which are shared among individuals of a given race. Furthermore, the familial aggregation and the resemblance between monozygotic twins that have been observed for anthropometric measurements of body fat distribution and for visceral AT accumulation measured by computed tomography, also suggest that genetic factors are involved in the determination of body fat distribution. Genetic susceptibility may also influence the relationship between visceral AT accumulation and the development of metabolic alterations. In this regard, it has been reported that the polymorphism of some genes (for example, the apolipoprotein (apo) E, apo B100 and lipoprotein lipase genes) is altering the relationship between visceral obesity and plasma lipoprotein-lipid levels. In conclusion, results presented in this paper suggest that genetic factors seem to have a significant influence on the propensity to accumulate AT in the visceral depot and that genetic factors also seem to affect the associations commonly reported between visceral obesity and the development of metabolic alterations. PMID- 9347401 TI - The non-genetic determinants of central adiposity. AB - Central adiposity carries an increased risk of non-insulin dependent diabetes mellitus (NIDDM), cardiac disease, hypertension and death, and is closely related to insulin resistance. Genetic factors explain a large proportion of the population variance in central adiposity, although the genotypic characteristics remain obscure. Hormonal factors such as endogenous sex steroid levels, the menopause, hormone replacement therapy and cortisol may influence body fat partitioning. The link between dietary factors and central adiposity is controversial, with contradictory results in the literature. Smoking is associated with lower total body fat, but investigations of its influence on central adiposity have also yielded contradictory results. Higher levels of physical activity are associated with lesser amounts of central fat, both cross sectionally and in intervention studies. Some of the contradictory results regarding putative influences on central adiposity may be due to limitations of some of the anthropometric parameters of central adiposity, such as the waist-hip ratio. Further research is required to clarify the relationships between many of these factors and with both compartments of central adiposity: subcutaneous abdominal and intraabdominal adipose tissue. PMID- 9347402 TI - Replacement of dietary fat by sucrose or starch: effects on 14 d ad libitum energy intake, energy expenditure and body weight in formerly obese and never obese subjects. AB - OBJECTIVE: To investigate the impact of a high-sucrose diet vs a high-starch and a high-fat diet on 14 d ad libitum energy intake, body weight, energy expenditure and sympathoadrenal activity. MEASUREMENTS: Food intake; body weight and composition (bioelectrical impedance); 24 h energy expenditure, substrate oxidation rates, spontaneous physical activity, heart rate and appetite sensations in a respiration chamber (VAS scores); plasma catecholamine concentration and blood pressure. SUBJECTS: Twenty normal-weight, healthy women, 9 post-obese (body mass index (BMI): 22.9 +/- 0.7 kg/m2) and 11 closely matched controls (BMI: 22.6 +/- 0.4 kg/m2). RESULTS: Average 14 d ad libitum energy intake was 13% and 12% lower on the starch diet compared with the sucrose and fat diets, respectively (P < 0.05). In both post-obese and normal-weight subjects, body weight and fat mass decreased significantly on the starch diet (by 0.7 +/- 0.2 kg and 0.4 +/- 0.1 kg, respectively, P < 0.05). No changes were observed on the fat or sucrose diets. After 14 d on the sucrose diet, 24 h energy expenditure as well as postprandial plasma adrenaline and noradrenaline concentrations, were significantly increased compared with the other two diets. Overall satisfy and palatability ratings were also highest on the sucrose diet. CONCLUSION: Intake of a 14-d ad libitum high-starch diet decreased energy intake and body weight compared with a high-fat or high-sucrose diet. The increased energy expenditure observed on the sucrose-rich diet can probably be explained both by the increased intake of energy and fructose (mainly from sucrose) on this diet. PMID- 9347403 TI - Psychological and metabolic responses of carbohydrate craving obese patients to carbohydrate, fat and protein-rich meals. AB - RATIONALE: A defective central serotonergic neurotransmission has been suggested to result in the concomitant occurrence of an appetite disorder and a disturbed mood. This syndrome was termed carbohydrate carving (CC) obesity. Excessive consumption of carbohydrate-rich snacks would, through a plasma amino acid mediated mechanism, restore serotonergic neurotransmission and thereby relieve the symptoms of atypical depression. OBJECTIVES: To test whether CC obese patients indeed exhibit symptoms of atypical depression, whether these symptoms can be alleviated by carbohydrate-rich snacks and whether they respond differently to the snacks than non-carbohydrate craving (NC) control subjects. Furthermore, we investigated whether differences between CC and NC patients could be related to peripheral metabolic differences. DESIGN: Double blinded, randomized with cross-over. Patients received three types of snacks (100/0/0, 70/29/1 and 35/3/62 energy percent carbohydrate/fat/protein respectively) on three consecutive test days. Before and after snack administration mood and performance were tested and blood samples were obtained. SUBJECTS: 9 CC and 17 NC obese patients, matched for sex, age and body mass index. MEASUREMENTS: Mood states (Profile of Mood States and Visual Analogue Scales) and performance (Bourdon-Wiersma cancellation test), serum glucose and insulin and plasma amino acid concentrations. RESULTS: Before snack consumption, CC patients had slightly higher anger and fatigue scores and tended to have lower mood scores than NC patients. The efficiency of performance increased in both groups after all snacks. No other psychological effects of the snacks were registered. Psychological and metabolic responses of CC and NC patients to the snacks were similar. CONCLUSION: Although they may have a somewhat disturbed mood, CC obese patients do not improve their mood states through ingestion of a carbohydrate rich snack. It seems, from a therapeutic point of view, useless to maintain the concept of carbohydrate craving. PMID- 9347404 TI - RU-486 (Mifepristone) ameliorates diabetes but does not correct deficient beta adrenergic signalling in adipocytes from mature C57BL/6J-ob/ob mice. AB - OBJECTIVE: To investigate the role of hypercorticism in the development of compromised beta-adrenergic signalling in adipocytes of mature C57BL/6J-ob/ob mice. DESIGN AND EXPERIMENTAL UNITS: Mature male ob/ob mice and their lean littermates were treated with vehicle or the specific glucocorticoid receptor (GR) antagonist, RU-486 (30 mg/kg bw/d) for 21 d. MEASUREMENTS: Blood glucose, serum insulin, adipocyte Glut-4 expression, adipocyte Gs alpha expression, adenylylcyclase activation by beta-adrenergic receptor (beta-AR) agonists in adipocyte membranes and mRNA levels for beta 1-, beta 2- and beta 3-adrenergic receptor subtypes in adipocytes. RESULTS: RU-486 reduced blood glucose levels in ob/ob mice to levels that were not different from lean mice. RU-486 also reduced serum insulin by approximately 50% in ob/ob mice, but failed to restore depressed Gs alpha or GLUT-4 expression in adipocytes of ob/ob mice. RU-486 produced a two fold increase in beta 3-AR mRNA in ob/ob mice and a small but significant improvement in isoprenaline-mediated adenylylcyclase activation. CONCLUSIONS: The present results indicate that glucocorticoid antagonism ameliorates diabetic symptoms of the mature ob/ob mouse, but does not lessen their obesity or fully reverse deficient expression and function of components of the adipocyte beta adrenergic signalling cascade. PMID- 9347405 TI - Changes in fat-free mass and fat mass in postpartum women: a comparison of body composition models. AB - OBJECTIVES: (1) To compare 2-, 3- and 4-component models of body composition based on total body water (TBW), underwater weighing (UWW), skinfold thicknesses (SF), total body potassium (TBK), dual-energy X-ray absorptiometry (DXA) and total body electrical conductivity (TOBEC); (2) to compare postpartum changes in body composition estimated by the 2-, 3- and 4-component models and (3) to test for an effect of pregnancy or lactation on the hydration, density and potassium content of fat free mass (FFM) in postpartum women. DESIGN: Longitudinal measurements of body composition at 3, 6 and 12 months postpartum. SUBJECTS: Thirty-five healthy postpartum women, aged 30.2 +/- 3.5 y. MEASUREMENTS: Body composition was estimated by 2-component models based on TBW, UWW, SF, TBK, DXA or TOBEC; 3-component models based on TBW and UWW (Fuller 3, Siri 3); and a 4 component model (Fuller 4) based on TBW, UWW and bone mineral content. RESULTS: Systematic differences were seen among the various body composition models, with the following ranking from lowest to highest estimate of fat mass (FM): TOBEC, TBW, Fuller 3, Siri 3, Fuller 4, UWW, SF, TBK, and DXA. Estimated changes in FFM and FM were not significantly different among methods, except for the 3-6 months FFM and FM changes estimated from TBW, which differed from SF, DXA, and TOBEC. Pregnancy-induced changes in the hydration, density and potassium content of FFM were not evident by 3 months postpartum (0.73 +/- 0.02, 1.099 +/- 0.015 kg/l and 2.31 +/- 0.10 g/kg, respectively). CONCLUSION: In spite of systematic differences among body composition models for the measurement of FFM and FM, changes in FFM and FM did not differ significantly among the models. Since there was no apparent effect of pregnancy or lactation on the postpartum composition of FFM, 2 component models of body composition are acceptable for use in postpartum women beyond the puerperium. PMID- 9347406 TI - Leptin serum levels in normal weight and obese children and adolescents: relationship with age, sex, pubertal development, body mass index and insulin. AB - OBJECTIVE: Leptin, the product of the ob gene, is present in higher concentrations in blood of obese subjects than of lean subjects. There is scarce information on the role of leptin in the pathogenesis of human obesity and little is known about leptin serum levels in obese children. DESIGN, SUBJECTS AND MEASUREMENTS: To evaluate the influences of age, sex, pubertal development and weight excess on serum leptin levels, we have studied 390 obese subjects (OS) and 320 normal weight subjects (NWS) aged 5-16 y. Fasting insulin concentrations were assayed in NWS, and an oral glucose tolerance test was carried out in OS and total insulin area under the curve (TIA) was calculated. RESULTS: Log-transformed values of leptin serum concentrations appeared to be distributed according to an acceptable Gaussian pattern. As observed in adults, serum leptin concentrations in children and adolescents were also increased (4-5 times) in OS as compared to NWS. In both males and females, subdivided according to pubertal stages, serum leptin varied significantly in stage IV-V as compared to the lower stages, with a reduction in males and an increase in females. On comparing the two sexes, greater serum leptin concentrations were observed in females of both NWS and OS. A significant linear correlation was found in both groups, subdivided according to sex and pubertal stage, between log values of serum leptin and standard deviation scores (SDS) of body mass index (BMI), and log-transformed relative body weight (RBW). Using partial correlation analysis in subjects subdivided according to sex and pubertal stages, log values of serum leptin and fasting insulin values, adjusted by age and SDS of BMI, correlated significantly with a weaker correlation in males than in females. In OS, the leptin concentrations correlated better with TIA than with fasting insulin. A weight reduction program (WRP) was carried out in 141 OS and significant reductions of serum leptin and fasting insulin were observed, showing a reduction of RBW. There was a correlation between the reduction of RBW and of serum leptin, but not of fasting insulin. No variation was found in non-responsive OS. RBW reduction correlated with leptin, but not with insulin (fasting and TIA), evaluated before the therapeutic program started. CONCLUSION: As observed in adults, obese children and adolescents have higher serum leptin concentrations. However, several conditions should be taken into account when evaluating leptin concentrations in children. There are differences, independent of BMI, relative to pubertal stage and sex, females having greater leptin concentrations than males. There is evidence of a possible role for leptin in the effectiveness of a weight reduction program in OS. PMID- 9347407 TI - Health care costs of obesity in New Zealand. AB - OBJECTIVE: To estimate the costs of health care that are attributable to obesity in New Zealand. METHODS: The 1991 health care costs of non-insulin dependent diabetes, coronary heart disease, hypertension, gallstone disease, post menopausal breast cancer and colon cancer were estimated and multiplied by the population attributable factor for obesity for each condition. The relative risk estimates were taken from the literature, the obesity prevalence from a 1990 New Zealand survey, and the costs and volumes of services were taken from a variety of sources and covered hospital (inpatient and outpatient) services, general practitioner consultations, pharmaceuticals, laboratory tests and ambulance services. Calculations were conservative and net of goods and services tax. RESULTS: A conservative estimate of the health care costs attributable to obesity for the six conditions was NZ$135 million. This represents about 2.5% of total health care costs which is similar to analyses from other countries. CONCLUSIONS: The health care costs of obesity as estimated are considerable. However, the total cost of overfatness to the New Zealand population is far greater than this because lesser degrees of overfatness, the health care costs of other obesity related conditions such as arthritis, the costs to individuals of weight-loss programs and the indirect and intangible costs were not included in the analysis. A substantial and wide-ranging public health effort is needed to turn around the increasing prevalence and costs of obesity. PMID- 9347408 TI - Insulinaemia and slight overweight: the case of Vietnamese hypertensives. AB - OBJECTIVE: To examine the relationship between hypertension, overweight and indices of insulin resistance in Vietnamese subjects. PATIENTS: One hundred and eight hypertensive subjects (51 men and 57 women) over 40 y of age were compared with 36 control subjects over 40 y of age. METHODS: Blood glucose and plasma insulin were measured at fasting and 2 h after 75 g glucose taken orally. RESULTS: Hypertensive subjects had significantly higher body mass index (BMI), triceps skinfold, waist and hip circumferences and waist/hip ratio. Glycaemia at fasting and after glucose were similar in the two groups. Insulinaemia at fasting and after glucose were significantly higher in the hypertensive subjects. In the whole series of hypertensive subjects, plasma insulin and insulin/glucose ratio at fasting and after glucose correlated significantly with BMI, triceps skinfold and waist and hip cicumferences. After exclusion of the subjects with BMI > 22 kg/m2, compared with the controls, plasma insulin and insulin/glucose ratio were significantly higher in the whole hypertensive group and separately in hypertensive men and women. The logistic regression analyses have shown that plasma insulin and insulin/glucose ratio at fasting and after glucose were significantly associated with hypertension, independently of gender, BMI and waist circumference. CONCLUSION: This study shows that in Vietnamese people, essential hypertension is associated with a significant increase in BMI, which however remains far lower than the definition threshold of occidental obesity and with a state of insulin resistance found despite very slight or no excess weight. PMID- 9347409 TI - Contrasting factors associated with abdominal and peripheral weight gain among adult women. AB - OBJECTIVE: To identify contrasts between the risk factors associated with abdominal weight gain and those associated with peripheral weight gain. DESIGN: Prospective mail survey. SUBJECTS: 44080 white, non-Hispanic, healthy women who were questioned in 1982 (baseline age 40-54 y) and 1992 about weight, diet, alcohol use, smoking, 10 physical activities and other variables. MEASUREMENTS: Self reports in 1992 identified 4261 women who gained weight in the abdomen and 7440 women who gained in the periphery (sites other than the abdomen). Using identical logistic models adjusted for age, baseline body mass index (BMI) and numerous covariates, the abdominal-gain group and the peripheral-gain group were separately compared with 10,888 women who did not gain weight. RESULTS: The likelihood of abdominal gain exceeded that of peripheral gain (by comparison of estimated odds ratios, abdominal vs peripheral) for high meat eaters (1.50 vs 1.15), frequent users of liquor (1.09 vs 0.54), moderate cigarette smokers (0.86 vs 0.59), heavy cigarette smokers (0.96 vs 0.36), cigarette quitters (2.13 vs 1.63), women with high parity (1.52 vs 1.15) and those who reported major weight gain since age 18 y (1.22 vs 0.65). Abdominal gain was less likely than peripheral gain for high vegetable eaters (0.71 vs 0.91), women who exercised > or = 4 h/wk [(especially aerobics/ calisthenics (0.28 vs 0.91) or walking (0.84 vs 1.06)], women who completed menopause (0.74 vs 0.98) and consistent users of estrogen replacement therapy (0.93 vs 1.22). CONCLUSION: A behavior or characteristic may be associated differently with the risks of abdominal and peripheral weight gain. This insight could strengthen recommendations for preventing major chronic diseases. PMID- 9347410 TI - Two decades of annual medical examinations in Japanese obese children: do obese children grow into obese adults? AB - OBJECTIVE: To investigate trends in frequency of obese children in Japan over two decades, the frequency of obese children who grow into obese adults and predictive factors for adult obesity. DESIGN: Annual cross-sectional studies for 22 y (1974-1995) with a follow-up study. SUBJECTS: Cross-sectional: Cumulatively 13,186 obese (% of standard body weight (SBW): > or = 120%) schoolchildren including 3158 extremely obese (> or = 140% of SBW) children out of 203,088 schoolchildren (age: 6-14 y) in Izumiohtsu City, Osaka, Japan. FOLLOW-UP: 151 initially obese children (initial age: 6-14 y and age at follow-up: 20-35 y) who lived in Izumiohtsu City. CONTROL: 3552 Japanese men and 4631 Japanese women (age: 20-35 y). MEASUREMENTS: Cross-sectional: height, weight, trunk circumference, skin-fold thickness, blood pressure and blood biochemicals. FOLLOW UP: height, weight, trunk circumference, skin-fold thickness during childhood, and body height and weight at follow-up. Adulthood obesity: > or = 120% of the average body mass indices (BMI) of the controls. RESULTS: Frequency of obese children increased from 5% to more than 10%, and that of extremely obese children increased from 1% to more than 2% during these 22 y. These increases were most prominent in the schoolboys aged 9-11 y. Prevalence of hyperglycemia and hyperlipidemia in the extremely obese children did not change, and that of hypertension and abnormal liver function gradually decreased during these two decades. After coming of age, 32.2% of the initially obese boys (relative risk: 5.3) and 41.0% of the initially obese girls (relative risk: 6.7) remained obese. BMI, percentage of the SBW and skin-fold thickness at the biceps during childhood were significantly larger in currently-obese girls. Positive correlations were demonstrated between these variables and percentage SBW at follow-up. CONCLUSIONS: Childhood obesity is increasing in Japan, especially in boys aged 9 11 y. Approximately 32% of the obese boys and 41% of the obese girls grow into obese adults, and the degree of obesity is a predictive factor for adult obesity. PMID- 9347411 TI - Small doses of triiodothyronine can change some risk factors associated with abdominal obesity. AB - OBJECTIVE: To elucidate whether the administration of small doses of triidothyronine (T3) can increase concentrations of sex hormone binding globulin (SHBG) in obese women with different types of obesity and to evaluate the potential metabolic benefits of such treatment. DESIGN: Daily administration of 20 micrograms of T3 during six weeks while maintaining habitual food intake and physical activity. SUBJECTS: Seventy premenopausal obese women (age: 41.2 +/- 1.5 y mean +/- s.e.m., body mass index (BMI): 34.4 +/- 0.7). MEASUREMENTS: Plasma concentrations of SHBG, lipids, insulin, thyroid hormones, sex hormones, blood glucose and insulin sensitivity (by euglycemic insulin clamp in 12 patients) at base line after six weeks of treatment. RESULTS: Six weeks treatment with small doses of T3 resulted in a significant increase in plasma SHBG. The increase of SHBG was higher in abdominal obesity and not associated with a significant change in body weight, plasma insulin concentration, insulin/glucose ratio of plasma insulin sensitivity (glucose disposal during insulin clamp). In patients with initially high SHBG the significant increase of insulin removal (as judged from the increase of c-peptide/insulin ratio) was observed. Treatment resulted in a reciprocal increase of T3, decrease of thyroxine (T4), and a more than double increase of T3/T4 ratio. CONCLUSIONS: Administration of small doses of T3 can increase the concentration of SHBG without changing insulin concentrations or sensitivity. As there was a significant decrease (by 36%) of T4 and parallel increase of T3 with a clear increase of T3/T4 ratio it seems possible that rather than lack of thyroid hormones a lower peripheral deiodination of T4 might be a factor contributing to the low SHBG concentration in abdominal obesity. Treatment with small doses of T3 may be considered to ameliorate some of the risk factors associated with abdominal obesity, particularly in some subgroups of obese women with a relative resistance to thyroid hormones possibly dependent on decreased peripheral deiodination of thyroxine. PMID- 9347412 TI - The use of multi-frequency impedance to determine total body water and extracellular water in obese and lean female individuals. AB - OBJECTIVE: To validate the assessment of total body water (TBW) and extracellular water (ECW) by multi-frequency bioelectrical impedance. SUBJECTS: Twenty-five overweight but otherwise healthy subjects and 20 lean subjects. DESIGN: Cross sectional. MEASUREMENTS: TBW and ECW were determined by dilution techniques. Prediction equations from the literature were used to calculate TBW and ECW from measured impedance at 100 and 50 kHz or 1 and 5 kHz, respectively. In 18 of the obese subjects, impedance was also measured with the electrodes placed at proximal sites. RESULTS: In lean and obese subjects, significant correlations were observed between the impedance index (H2/Z) at high frequencies with TBW (r = 0.90, P < 0.001 in lean and r = 0.80, P < 0.001 in obese subjects) and at low frequencies with ECW (r = 0.87, P < 0.001 and r = 0.77, P < 0.001 respectively). Proximal placement of electrodes slightly improved the correlation between the impedance index and TBW (from r = 0.83 to r = 0.90 at 50 kHz and from r = 0.85 to r = 0.90 at 100 kHz) and ECW (from r = 0.77 to r = 0.83 at 1 kHz and from r = 0.79 to r = 0.85 at 5 kHz). The association of ECW and TBW with H2/Z was different for obese and lean subjects: in obese subjects a given amount of TBW or ECW corresponded with a lower index. An equation consisting only of the impedance index could predict TBW and ECW with small mean errors in lean (1.3 and 0.8 kg respectively) and obese subjects (0.1 and 0.0 kg respectively). Applying a more specific equation, including other subject characteristics, resulted in larger prediction errors in obese subjects, illustrating the population specificity of prediction equations. Furthermore an association was observed of the prediction bias of TBW and ECW with TBW (r = 0.48, P < 0.01) and ECW (r = 0.70, P < 0.001) respectively, and with body water distribution (r = -0.38 and r = 0.33 respectively, P < 0.05). TBW and ECW were also associated with weight (r = 0.76 and r = 0.71 respectively, P < 0.001) and body mass index (BMI) (r = 0.54 and r = 0.53 respectively, P < 0.001). CONCLUSION: It appeared from this study that the accuracy of TBW and ECW estimation with the impedance technique is dependent on the absolute amount of TBW and ECW. A higher amount of TBW and ECW in obese subjects may contribute to a difference in prediction error between lean and obese individuals. PMID- 9347413 TI - Familial risk ratios for extreme obesity: implications for mapping human obesity genes. AB - OBJECTIVE: To determine familial risk ratios for extreme obesity to aid in the design of obesity linkage studies. DESIGN: Family study of obesity. SUBJECTS: 2349 first-degree relatives (parents and siblings) of 840 probands who are members of the National Association to Advance Fat Acceptance (NAAFA) and 5851 participants of the first phase of the National Health and Nutrition Examination Survey III. METHODS: Computed age-gender standardized risk ratios (SRRs) for obesity in relatives categorized by the level of obesity in the index case (proband). MEASUREMENT: Body mass index (BMI) (kg/m2). RESULTS: The risk of extreme obesity (BMI > or = 40) in relatives of extremely obese women (BMI > or = 40) was more than five times greater than in the population; furthermore, the risk of obesity in relatives was approximately linearly associated with the degree of obesity in the proband. The risk of thinness in relatives of obese individuals was substantially lower than in the general population. CONCLUSION: Because the familial risk ratio for extreme obesity is higher than for moderate levels of obesity, the number of families required to achieve adequate statistical power in gene mapping studies of obesity can be reduced substantially by focusing on family members of extremely obese individuals (BMI > or = 40). PMID- 9347414 TI - A meta-analysis of the past 25 years of weight loss research using diet, exercise or diet plus exercise intervention. AB - OBJECTIVE: The therapeutic effectiveness of diet, exercise, and diet plus exercise for weight loss in obesity was determined. DATA SOURCES: All human research reported in English, published in peer-reviewed scientific journals within the past 25 y was reviewed. STUDY SELECTION: Acceptance criteria (n = 493 from > 700 studies) were that a therapeutic intervention of diet, exercise or diet plus exercise was employed, specifically for weight reduction in obese adult humans and that weight change was reported numerically. Only aerobic exercise studies were included, while drug, hormone and surgical treatments were excluded. DATA EXTRACTION: All data were extracted by the same investigator from the original research report. Except for gender and program type, all extracted data were numerical. DATA SYNTHESIS: ANOVA, with a Newman-Keuls post hoc test, was used to determine differences among programs (P < 0.05). One analysis was performed on the group mean data and one based on effect sizes. Analyses were repeated using initial body weight, initial percent body fat and program length, as covariates. RESULTS: Primarily, subjects aged 40 y have been studied (39.5 +/- 0.4 y, mean +/- s.e.m.) who are only moderately obese (92.7 +/- 0.9 kg, 33.2 +/- 0.5 body mass index (BMI), 33.4 +/- 0.7% body fat); for short durations (15.6 +/- 0.6 weeks). Exercise studies were of a shorter duration, used younger subjects who weighed less, had lower BMI and percentage body fat values, than diet or diet plus exercise studies. Despite these differences, weight lost through diet, exercise and diet plus exercise was 10.7 +/- 0.5, 2.9 +/- 0.4* and 11.0 +/- 0.6 kg, respectively. However, at one-year follow-up, diet plus exercise tended to be the superior program. Effect size and covariate analyses revealed similar program differences. CONCLUSION: Weight loss research over the past 25 y has been very narrowly focused on a middle age population that is only moderately obese, while the interventions lasted for only short periods of time. The data shows, however, that a 15-week diet or diet plus exercise program, produces a weight loss of about 11 kg, with a 6.6 +/- 0.5 and 8.6 +/- 0.8 kg maintained loss after one year, respectively. PMID- 9347416 TI - Risch's lambda values for human obesity estimated from segregation analysis. PMID- 9347415 TI - Association of intraabdominal fat and carotid atherosclerosis in non-obese middle aged men with normal glucose tolerance. AB - OBJECTIVE: To investigate the association of visceral fat accumulation and carotid atherosclerosis in non-obese men with normal glucose tolerance. SUBJECTS: Ninety eight middle-aged men with normal glucose tolerance who had a body mass index (BMI) < 25 kg/m2 (mean age: 52.6 y, mean BMI: 22.5 kg/m2). METHOD: Using ultrasonography, we simultaneously measured the abdominal fat distribution and intima-media thickness (IMT) of the carotid artery wall. RESULTS: Pmax, an index of the absolute amount of visceral fat, was found to be significantly correlated with IMT and fasting Insulin concentration (r = 0.37, P < 0.005 and r = 0.54, P < 0.0001). Abdominal wall fat index (AFI), the ratio of maximum thickness of the preperitioneal fat and the minimum thickness of the subcutaneous fat, was not correlated with IMT. CONCLUSION: Visceral fat accumulation may play a role in the progression of atherosclerosis even in non-obese middle-aged men. PMID- 9347417 TI - The language of compliance: health policy and clinical practice for the severely mentally ill. AB - This paper appraises the issues surrounding 'compliance' from a biomedical and specifically psychiatric perspective with particular reference to the drug treatment of psychiatric patients and the influential factors in the development of health policy. I examine how existing conceptualisations fail to have a significant impact on the way people with severe mental illness (chronic disorders such as schizophrenia) interact with prescribed medication (usually neuroleptic) and explore what potential insights can be gleaned from the anthropological literature. PMID- 9347418 TI - Reform and reshaping mental health services in the Montreal area. AB - Over the past 40 years, the mental health care system has been radically transformed from one focused on institutionalized care to one centred on treatment in community settings. Following the example of the UK and USA, Quebec is travelling down a similar path in transferring mental illness services from mental hospitals to the community. The purpose of this present study is firstly to examine why and how this administrative settlement has been reached; secondly to explain the operation of the service, especially in both sectors where Quebec has innovated: family housing and individualized service plans. PMID- 9347419 TI - A comparison of public attitudes in Britain and Saudi Arabia towards auditory hallucinations. AB - BACKGROUND: The successful introduction of community interventions is partly dependent on public beliefs about the aetiology and treatment of psychiatric difficulties and tolerance of community integration. METHOD: This study examined community attitudes towards auditory hallucinations in Saudi Arabia (SA) and the United Kingdom (UK) concerning (a) causes of auditory hallucinations, (b) the efficacy of interventions and (c) levels of social rejection. RESULTS: Responses from 281 patients attending their general practitioners indicated that those living in Saudi Arabia were most likely to believe that hallucinations are caused by Satan or due to magic, while the UK sample were more likely to cite schizophrenia or brain damage. While the Saudi sample believed that religious assistance would be most effective, the UK sample supported medication and psychological therapies. Beliefs about aetiology and treatment were unrelated to educational attainment. There was a greater degree of social rejection of patients in Saudi Arabia, but here educational attainment was of significance. CONCLUSIONS: These results suggest that beliefs about aetiology are related to treatment recommendations and social distancing, and thus have implications for the care of Arabic patients living in Western countries as well as for the use of Western interventions in non-Western cultures. PMID- 9347420 TI - A cross-cultural study of the attitudes of mental health professionals towards auditory hallucinations. AB - The usefulness of psychological interventions for auditory hallucinations is becoming increasingly accepted in Western cultures, but there are few data concerning the views of professionals working in non-Western societies. In this study, 195 psychologists and psychiatrists working in Saudi Arabia (SA) and Britain (UK) responded to a questionnaire regarding their (a) attitudes towards various clinical aspects of auditory hallucinations, (b) perceptions of the clinical value of psychological and pharmacological treatments and of the inputs of the two professions and (c) levels of social distance from people who experience auditory hallucinations. UK staff believed that there is a greater range of possible causes and diagnoses for auditory hallucinations than SA staff, who in turn had more confidence in the efficacy of psychological and pharmacological treatments. UK staff reported significantly less social distance from this group of patients. The results suggest that the use of psychological approaches to helping people with auditory hallucinations could be affected by cultural views of the causes and treatment. PMID- 9347421 TI - Crisis beds: the interface between the hospital and the community. AB - The main role of the crisis unit at a state psychiatric hospital was found to be the provision of brief hospitalisation for patients with adjustment disorders and personality disorders. The four bed crisis unit treated 14% of all patients admitted to the hospital, with an average length of stay less than three days. A study of 78 crisis unit patients found that 77% were able to be discharged directly to the community, and only 18% were readmitted during the following six months. PMID- 9347422 TI - Family and cultural correlates of depression among Chinese elderly. AB - This study hypothesized that depressive experiences of the elderly could be aggravated by universal factors such as low social status, poor health, financial strain, and unhealthy lifestyle, as well as by factors specific to an indigenous socio-cultural environment (stressful family dynamics) of a given population. Three hundred and fifty Chinese subjects aged 65 or older were interviewed either at their homes or in the geriatric out-patient clinic of Beijing Hospital. Hierarchical logistic regression was used to examine significant predictors of depression. Results showed that certain social status, poor physical health, financial strain, unhealthy lifestyle, and stressful family situation explained 47 percent of the variance in depression. However, stressful family situation alone explained 13 percent of the variance in depression, indicating that family factors were important predictors of depression for Chinese elderly. Furthermore, this study demonstrated for the first time that verbal abuse within Chinese families is a significant correlate of depression among the elderly. Cultural implications of these findings are discussed. PMID- 9347423 TI - Community psychiatry in Hong Kong. AB - Community psychiatry is well developed in many western countries. However, this psychiatric subspecialty has only recently been officially recognized and established in Hong Kong. This article describes the development and current scope of services. It illustrates how local psychiatrists have met the challenge of adopting a western service model to suit the local Chinese population, with its different socio-cultural value system. PMID- 9347424 TI - Post partum psychosis: a clinical study. AB - In this study of 192 cases of post partum psychosis, the mean age of cases was 24.2 years. A past history of post partum psychosis was present in 16 cases (8.3%). As per the RDC categories, a majority of patients had unspecified functional psychosis and developed psychosis after the birth of first child. There was a positive correlation between the birth of female child and psychosis. The majority of cases developed psychosis within first 2 weeks after delivery. There were several other statistically significant differences when these cases were compared with non-puerperal, disease-matched controls. PMID- 9347425 TI - Dropping out from child psychiatric treatment: reasons and outcome. AB - This study explores the reasons underlying dropping out from a child psychiatric clinic in Hong Kong, and the outcome of these children. A reluctance to accept psychiatric help, or the possibility of being labelled as psychiatrically ill, and differences between the families and doctors in their perception of the presenting problems are common reasons given for dropping out. A substantial number of children who dropped out continued to have some degree of psychiatric morbidity two years later. These results highlight the need to increase the community's awareness of the nature of child psychiatry, and for doctors to be sensitive to the families' perception of the presenting problems so as to minimise the likelihood of dropping out. PMID- 9347426 TI - HIV disease and anesthesiologists' risk: can old dogs learn new tricks? PMID- 9347427 TI - Compliance with gloving in anesthesia: an observational study of gloving practice at induction of general anesthesia. AB - STUDY OBJECTIVE: To gather direct observational data on anesthesiologists' compliance with universal precautions' gloving standards during induction of general anesthesia. DESIGN: Prospective, observational study. SETTING: Operating theaters of an Israeli government teaching hospital. SUBJECTS: Over a four-month period, all "first case of the day" general anesthetics were observed to determine if the anesthesiologist directly administering patient care wore gloves during the period of anesthetic induction. All anesthesia department members were observed and none was aware of the ongoing study. MEASUREMENTS AND MAIN RESULTS: Resident anesthesiologists were found to be more compliant with gloving policy than their attendings (61.8% vs. 33.7%, p < 0.0001). However, the lower compliance among the attendings was entirely attributable to the most senior staff members (over age 55 years) whose compliance rate was 11.5% versus 55.6% for attending staff below age 55 years (p < 0.0001). Departmental compliance as a whole was 49.6%. Compliance in pediatric cases averaged 10% and was equally poor among all department staff. CONCLUSIONS: Although glove use remains inconsistent, in less than one and one half years since institution of a departmental gloving policy, a substantial degree of compliance was achieved. Nevertheless, further efforts are still needed to improve compliance with universal precautions. In this study, glove use was particularly deficient in pediatric cases and among senior staff aged 55 years and older. Pinpointing specific areas of greatest deficiency may prove useful in guiding additional efforts to improve compliance with universal precautions. PMID- 9347428 TI - Remifentanil versus alfentanil in a balanced anesthetic technique for total abdominal hysterectomy. AB - STUDY OBJECTIVES: To compare the intraoperative effects and recovery characteristics of remifentanil hydrochloride and alfentanil when administered as part of balanced anesthesia, and to assess the effects of an additional remifentanil infusion administered as analgesic pretreatment before removal of the uterus. DESIGN: Multicenter, double-blind, randomized, parallel-group study. SETTING: Two university hospitals. PATIENTS: 35 ASA physical status I, II, and III women scheduled for elective total abdominal hysterectomy with general endotracheal anesthesia. INTERVENTIONS: Patients were premedicated with midazolam 0.05 mg/kg intravenously (i.v.). Anesthesia was induced with thiopental 2 mg/kg, vecuronium 0.15 mg/kg, and a single dose of opioid over 60 seconds (Pump 1): remifentanil 2 micrograms/kg (Remi/Placebo and Remi/Remi groups) or alfentanil 50 micrograms/kg (Alf/Placebo group). Anesthesia was maintained with a nitrous oxide/oxygen mixture (66:34 ratio) and a continuous opioid infusion: remifentanil 0.25 microgram/kg/min (Remi/Placebo and Remi/Remi) or alfentanil 0.5 microgram/kg/min (Alf/Placebo). At skin incision, a second blinded drug infusion was also initiated (Pump 2): remifentanil 0.25 microgram/kg/min (Remi/Remi) or saline placebo (Remi/Placebo and Alf/Placebo). Intraoperative responses were controlled with single doses of opioid and/or rate titrations via Pump 1. Pump 2 was terminated on removal of the uterus. Pump 1 was terminated at skin closure. MEASUREMENTS AND MAIN RESULTS: The mean (+/- SD) opioid infusion rates administered for the duration of Pump 2 to suppress responses to removal of the uterus were 0.49 +/- 0.27 microgram/kg/min, 1.99 +/- 1.34 micrograms/kg/min, and 0.49 +/- 0.07 microgram/kg/min for the Remi/Placebo, Alf/Placebo, and Remi/Remi groups, respectively. At these rates, similar proportions of patients in the Remi/Placebo (67%) and the Alf/Placebo (60%) groups had responses. Fewer patients had responses in the Remi/Remi group (8%) compared with the Remi/Placebo and Alf/Placebo groups (p < 0.05). The mean total opioid doses used during maintenance were 84.6 micrograms/kg (Remi/Placebo), 393 micrograms/kg (Alf/Placebo), and 68.7 micrograms/kg (Remi/Remi). Awakening times were significantly shorter (p < 0.05) in the remifentanil population compared with the alfentanil population, but discharge times were similar. More patients received naloxone to reverse opioid effects in the alfentanil population (60%) than in the remifentanil population (20%) (p < 0.05). CONCLUSIONS: A mean remifentanil infusion of 0.49 microgram/kg/min is as effective as a mean alfentanil infusion of 1.99 micrograms/kg/min in suppressing intraoperative responses. Doubling of the remifentanil infusion to 0.5 microgram/kg/min before the major stress event improves suppression of responses and lowers intraoperative use of remifentanil without prolonging recovery times. Remifentanil allows faster awakening times than alfentanil, but preemptive administration of postoperative analgesics is recommended to facilitate discharge. PMID- 9347430 TI - Preoperative pregnancy testing: a survey of current practice. AB - STUDY OBJECTIVE: To examine contemporary practices and opinions regarding preoperative testing requirements, with special emphasis on perioperative pregnancy recognition and consequences thereof. DESIGN AND SETTING: Anonymous questionnaire survey distributed to 300 (almost exclusively American) physicians attending the 1996 Society of Obstetric Anesthesia and Perinatology meeting. MEASUREMENTS AND MAIN RESULTS: Responses from 169 anesthesiologists indicated that approximately one-third mandated pregnancy testing via departmental policy. More anesthesiologists (p = 0.02) mandated routine pregnancy testing of all elective (30%) versus all emergency (17%) surgical patients. Sixty-six percent versus 20% percent, respectively, would require rather than simply offer pregnancy testing when history indicated possible pregnancy; 20% and 15%, respectively, of those surveyed indicated elective surgery would be canceled by the anesthesiologist if the patient were pregnant or refused testing (p = NS). Although 98% of respondents recognized a legal requirement to report child abuse, and 82% believed pregnancy in a juvenile (i.e., the child's age is under local legal defined age for consent to sex) by definition constituted child abuse, fewer than 4% would report this information to the police, even if the impregnating male were known to be an adult. CONCLUSIONS: The desire to identify pregnancy using patient history was most prevalent among anesthesiologists, with less than one third using mandatory, departmentally imposed screening programs. Positive test results in minors are shared primarily with surgeons and patients, occasionally with parents and social services, but rarely with police, although a positive test almost universally signified child abuse, and mandatory reporting laws were acknowledged by anesthesiologists surveyed. PMID- 9347429 TI - Effectiveness of bupivacaine administered via femoral nerve catheter for pain control after anterior cruciate ligament repair. AB - STUDY OBJECTIVE: To evaluate the quality of pain control achieved with continuous local anesthetic infusion via a femoral nerve catheter, and to determine the optimum concentration of bupivacaine necessary to maintain pain control after full surgical anesthesia is established with 0.5% bupivacaine. DESIGN: Randomized, prospective study. SETTING: Tertiary care teaching center. PATIENTS: 25 ASA physical status I and II patients scheduled to undergo arthroscopically aided anterior cruciate ligament (ACL) reconstruction by one surgeon, and who were willing to accept a femoral nerve catheter for postoperative pain control. INTERVENTIONS: All patients received general anesthesia with propofol/alfentanil (10 ml/1 ml) mixture and nitrous oxide/oxygen (60%/40%) mixture via endotracheal tube. After induction of general anesthesia, a femoral nerve catheter was inserted with the aid of a nerve stimulator, and 20 ml of 0.5% bupivacaine was administered. The surgery was completed in a standard manner and the patients were randomized into three groups for the concentration of local anesthetic to continue the pain relief into the recovery phase. On awakening, all patients were determined to have a functioning femoral nerve catheter. Group 1 received 0.0625% (n = 8) bupivacaine, Group 2 0.125% (n = 9) bupivacaine, and Group 3 0.25% (n = 8) bupivacaine; all doses were initiated in a blinded manner at 0.12 ml/kg/hr. Patients also received intravenous patient-controlled analgesia with morphine via demand mode only, with a 1.0 mg dose and a 6 minute lock-out interval. MEASUREMENTS AND MAIN RESULTS: Pain was determined at defined intervals by visual analog scale (VAS). Data collected included demographics, VAS scores, and total morphine administered. All patients were pain-free on emergence from general anesthesia. No patient required parenteral opioid for pain control while in the postanesthesia care unit. There were no significant differences in pain scores among groups, and average pain scores (2.5 to 4.0) indicate good pain control throughout the entire hospitalization. There were no complications. CONCLUSIONS: Low concentrations of bupivacaine delivered via femoral nerve catheter after an established femoral nerve block can provide excellent postoperative pain control after ACL reconstruction. PMID- 9347432 TI - Total-body oxygen consumption after isoflurane anesthesia: effects of mild hypothermia and combined epidural-general anesthesia. AB - STUDY OBJECTIVES: To determine the effects of epidural anesthesia and avoidance of intraoperative heat loss on the increase in total-body oxygen consumption in the immediate postoperative period after major intraabdominal surgery. DESIGN: Prospective, randomized (with regard to temperature management) study. SETTING: University medical center. PATIENTS: 24 ASA physical status I, II, and III adults. INTERVENTIONS: All patients received either isoflurane-nitrous oxide (N2O)-opioid general anesthesia or combined epidural-general anesthesia; patients were randomly assigned to active intraoperative warming or routine thermal care. MEASUREMENTS AND MAIN RESULTS: VO2 was measured by indirect calorimetry preoperatively (T0), immediately postoperatively (T1), and 60 to 90 minutes later (T2). For all patients, VO2 was 57 +/- 45% (mean +/- SD) greater at T1 than at T0 (p < 0.05). After isoflurane-N2O-opioid general anesthesia, VO2 increased 15 +/- 20% in normothermic patients (core temperature, 36.4 +/- 0.2 degrees C) compared with 69 +/- 52% in hypothermic patients (35.0 +/- 0.5 degrees C). After combined epidural-general anesthesia, VO2 increased 86 +/- 39% on emergence in normothermic (36.4 +/- 0.2 degrees C) and 58 +/- 11% in hypothermic (35.1 +/- 0.4 degrees C) patients. CONCLUSIONS: Total-body VO2 was increased in the immediate postoperative period. After general anesthesia, the magnitude of the increase in VO2 was significantly less in normothermic patients than in hypothermic patients. After combined epidural-general anesthesia, VO2 was increased in normothermic and in hypothermic patients. PMID- 9347431 TI - A randomized, double-blind, placebo controlled comparison of droperidol, ondansetron, and metoclopramide for the prevention of vomiting following outpatient strabismus surgery in children. AB - STUDY OBJECTIVE: To compare the efficacy of ondansetron, droperidol, or metoclopramide with placebo in preventing postoperative vomiting following strabismus surgery. STUDY DESIGN: Randomized, double-blind, placebo-controlled clinical trial. SETTING: University outpatient surgery center. PATIENTS: 160 ASA physical status I and II children ages 1 to 12 years who were scheduled for strabismus surgery. INTERVENTIONS: Administration of either ondansetron 100 mcg/kg, metoclopramide 250 mcg/kg, droperidol 75 mcg/kg, or placebo intravenously after induction of anesthesia. MEASUREMENTS AND MAIN RESULTS: Both ondansetron and droperidol were superior to metoclopramide and placebo in preventing predischarge vomiting, with incidences of 5%, 5%, 32%, and 25%, respectively. However, there was no difference in the incidence of postdischarge vomiting among the groups (ondansetron 25%, droperidol 25%, metoclopramide 20%, and placebo 25%). CONCLUSIONS: While both ondansetron and droperidol are more effective than metoclopramide when compared with placebo in decreasing the incidence of predischarge vomiting, none of these drugs was more effective than placebo in decreasing the incidence of postdischarge vomiting. Recovery from anesthesia was not significantly different among the groups as assessed by time to awakening, initial Steward score, and time to discharge. PMID- 9347434 TI - Preoperative laboratory testing in children undergoing elective surgery: analysis of current practice. AB - STUDY OBJECTIVE: To evaluate current practice in preoperative testing of healthy children undergoing elective surgery that is not expected to result in significant blood loss. DESIGN: Survey of members of the Society for Pediatric Anesthesia. SETTING: Anesthesiologists practicing in North America. POPULATION: A total of 1,200 questionnaires were mailed. INTERVENTIONS: Questionnaires were mailed to all members of the Society for Pediatric Anesthesia. All members were asked to specify which tests were routinely performed and to state why. Specific questions were asked about performing complete blood count (CBC), hemoglobin (Hb), hematocrit (Hct), and urine analysis (UA) in all patients, pregnancy test in adolescents, prothrombin time (PT) and activated partial thrombin time (PTT) prior to tonsillectomy, and sickle cell testing in black and/or Mediterranean children. MEASUREMENTS AND MAIN RESULTS: 685 of 1,200 (57%) questionnaires were returned. No attempt was made to identify and follow-up with nonresponders. Hb testing is routinely performed in 27% to 48% of the children depending on the age of the patient. UA is ordered preoperatively in less than 15% of the children. Pregnancy test was ordered by 43% of the respondents. Hemostatic tests prior to tonsillectomy were conducted by 45% of the anesthesiologists. CONCLUSION: The results indicate the present practice of routine preoperative laboratory testing for children undergoing elective outpatient surgery. In spite of the many studies that indicate no specific benefits of performing routine preoperative testing in healthy children undergoing scheduled surgery, many physicians continue to order these tests in all such children. PMID- 9347433 TI - Comparison of 0.25% ropivacaine and bupivacaine for epidural analgesia for labor and vaginal delivery. AB - STUDY OBJECTIVE: Part 1: To measure ropivacaine levels in the mother and infant at delivery after continuous lumbar epidural infusion. Part 2: To compare epidural ropivacaine to epidural bupivacaine for labor analgesia in regard to effectiveness, motor blockade, and maternal and neonatal effects. DESIGN: Part 1: Open-labelled, non-blind study. Part 2: Randomized, double-blind study. SETTING: Labor and delivery units of two academic hospitals. PATIENTS: Part 1: 20 ASA physical status I and II parturients in active labor. Part 2: 81 ASA physical status I and II parturients in active labor. INTERVENTIONS: For Part 1, 8 to 12 ml of 0.25% ropivacaine was administered through a lumbar epidural catheter to achieve a T10 dermatomal sensory level. An infusion of 0.25% ropivacaine, 8 to 10 ml/hr, maintained this sensory level. Maternal and umbilical cord blood samples obtained at delivery were analyzed for ropivacaine concentration. For Part 2, anesthetic management was similar to that previously described except patients were randomized to receive either 0.25% ropivacaine or 0.25% bupivacaine. Onset, regression, maximal spread of sensory block, and onset and degree of motor blockade were measured. Contraction pain as assessed using a visual analog scale (VAS), maternal blood pressure, and heart rate were determined every 5 minutes until a stable VAS-contraction score was achieved, and every 30 minutes thereafter. Neonatal assessment included Apgar scores and neurologic and adaptive capacity scores (NACS) at 15 minutes, 2 hours, and 24 hours. MEASUREMENTS AND MAIN RESULTS: For Part 1, the total and free maternal arterial concentrations of ropivacaine at delivery were 0.64 +/- 0.14 microgram/ml and 0.10 +/- .02 microgram/ml, respectively; the umbilical venous total and free concentrations were 0.19 +/- 0.03 microgram/ml and 0.12 +/- 0.07 microgram/ml, respectively (n = 12). The umbilical arterial and venous concentrations did not differ for both the free and total concentrations. For Part 2, there was no difference between ropivacaine and bupivacaine in the variables measured. Umbilical cord gases and Apgar scores were not different between the two groups; NACS were higher at 15 minutes and 2 hours in the ropivacaine group (p < 0.05) than the bupivacaine group. CONCLUSION: Both ropivacaine and bupivacaine produced excellent analgesia for labor with no major adverse effect on the mother or neonate. PMID- 9347435 TI - Intubation in children after 0.3 mg/kg of mivacurium. AB - STUDY OBJECTIVE: To distinguish among potential predictors of early, easy intubation in children, including apnea, neuromuscular block at two sites, and time, after administration of 0.3 mg/kg of mivacurium. DESIGN: Prospective, randomized study. SETTING: Operating rooms of Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania. PATIENTS: 60 ASA physical status I and II children aged 2 through 7 years, scheduled for elective surgical procedures requiring endotracheal intubation. INTERVENTIONS AND MEASUREMENTS: After premedication with midazolam, general anesthesia was induced with halothane and nitrous oxide, and patients were randomly assigned to one of four groups. Mivacurium 0.3 mg/kg was given and tracheal intubation was begun 45 seconds after its injection, or when apnea, block of the orbicularis oculi, (OO) or block of the adductor pollicis (AP) was noted. Intubation conditions were evaluated by an experienced endoscopist. MAIN RESULTS: The first clinical event after administration of mivacurium 0.3 mg/kg was apnea at 43 seconds (median) (average 48 seconds, SEM 2 seconds) after injection. The difference in the time at which neuromuscular block occurred at the AP (median 75 seconds) (average 77 seconds, SEM 2 seconds) and the OO (median 63 seconds) (average 68 seconds, SEM 4 seconds) was statistically, but not clinically, significantly different. All nine intubations that were begun at least 90 seconds after administration of mivacurium resulted in good or excellent intubation conditions, as did 30 of the 51 intubations started earlier. CONCLUSIONS: In children, there is no advantage to monitoring neuromuscular function at the OO rather than the AP. After administration of 0.3 mg/kg of mivacurium, a 90-second interval before the start of intubation was a better predictor of good intubation conditions during halothane anesthesia (1% inspired) than were changes in evoked neuromuscular function. PMID- 9347436 TI - Comparison of respiratory effects of tramadol and oxycodone. AB - STUDY OBJECTIVE: To compare the respiratory effects of tramadol and oxycodone. DESIGN: Placebo-controlled, double-blind randomized study. SETTING: IV Department of Surgery, Helsinki University Central Hospital. PATIENTS: 36 ASA physical status I and II patients undergoing minor surgery with general anesthesia. INTERVENTIONS: The respiratory effects of intravenous (i.v.) tramadol 0.6 mg/kg and oxycodone 0.04 mg/kg were compared after induction of anesthesia with propofol and succinylcholine-facilitated endotracheal intubation. Patients spontaneously breathed halothane in 70% nitrous oxide and oxygen via a nonrebreathing valve. The trial drugs or placebo were given after recovery from neuromuscular block. MEASUREMENTS AND MAIN RESULTS: Inspiratory and expiratory oxygen and end-tidal carbon dioxide concentrations (ETCO2), tidal volume (VT) minute volume of ventilation (VE), and respiratory rate (RR) were recorded by side-stream spirometry with end-tidal halothane of 0.3% for 30 minutes before surgery. Oxycodone caused a significant respiratory depression seen as an increase in the inspiratory-expiratory oxygen difference and ETCO2 and as a decrease in VE and RR. On the contrary, the effect of tramadol were similar to those of placebo. VT was not affected by any study drug. CONCLUSION: Tramadol was not associated with respiratory depression in the present setting. PMID- 9347437 TI - Use of a J-wire cover as an endotracheal tube changer. PMID- 9347438 TI - A case of artifactual rebreathing. PMID- 9347439 TI - False low pulse oximetry reading associated with the concomitant use of a peripheral nerve stimulator and an evoked-potential stimulator. AB - One of the sources of error in pulse oximetry readings is associated with an abnormal signal-to-noise ratio. The pulse oximeter distinguishes the light absorbance of arterial blood from that of other absorbers by differentiating between a constant component and a pulsating component. The pulsating component is almost exclusively the result of arteriolar bed pulsations. Because pulse oximetry is based on the assumption that arterial blood is the only pulsatile absorber, any other fluctuating phenomenon could constitute a source of error. We report a case in which a low pulse oximetry reading was associated with concomitant use of a pulse oximeter and a peripheral nerve stimulator on the same arm. Further tests conducted using a nerve stimulator and a sensory evoked potential stimulator with different amplitudes and frequencies confirmed the association and delineated the relationship between frequency and amplitude of stimulation and the degree of artificial desaturation. A theoretical explanation for this phenomenon is presented. PMID- 9347442 TI - Understanding the term "cost-effectiveness". PMID- 9347441 TI - Long-term skin hyperpigmentation after electrocardiographic monitoring. PMID- 9347443 TI - Emergency transtracheal jet ventilation in high grade airway obstruction. PMID- 9347440 TI - Blood, bone, and dura: anesthesia responsibility and pediatric neurosurgery. AB - In conclusion, providing anesthesia for a small child undergoing craniofacial reconstructive surgery is an enormous challenge. Even with the most experienced pediatric anesthesiologist and pediatric surgeons, problems can develop suddenly and lead, as they did in this case, to serious morbidity and even death. It is difficult to determine whether the anesthesiologists' "success" in this case in warding off a malpractice verdict was due to their lawyer's ability to convince the court they delivered a level of "care ordinarily supplied by physicians in their specialty," or, rather, due to the fact that defense experts were more convincing than those of the plaintiffs. Regardless, I do not think there were any "winners" in this situation. PMID- 9347444 TI - A lower solubility recommends the use of desflurane more than isoflurane, halothane, and enflurane under low-flow conditions. PMID- 9347445 TI - Inhalation sedation/analgesia with sevoflurane. PMID- 9347446 TI - Managed care in the United States. AB - Medical care in the United States continues to consume an increasing amount of the Gross Domestic Product. To control the rising costs of health care many industries have turned to a controlled form of financing and delivery of health care--often referred to as managed care. Many types of managed care exist, including preferred provider organizations (PPO), exclusive provider organization (EPO), and health maintenance organizations (HMO). HMOs involve prepaid premiums, limited panels of providers and assumption of financial risk on the part of the providers. A variety of HMOs are currently operating in the United States. Managed care involves taking risks by those who administer it. Some methods of controlling patient and physician behaviour by taking risks are capitation, risk pools and withholds. With capitation the physician is paid a 'per member per month' fee regardless of whether the patient uses the service. Risk pools are concerned with who shares the risk; for example, the primary physician shares the financial risk with specialists. Withholds involve a fee-for-service with a portion withheld which may be returned to the provider if he/she is parsimonious. A concern expressed about HMOs is the possibility of restricted services. Moreover, hospital expenses make up a large portion of the total health care dollar. In 1995 the average length of stay for a Medicare patient was 6.1 days as opposed to 3.9 days for the non-Medicare patient. Indeed, HMOs were the leaders in the development of same-day surgery and out-patient treatment. Increasingly, in the United States, public and social insurance plans are turning to managed care as a method to control health care expenditure. Some government insurance plans, such as Medicare and Medicaid, also increasingly offer managed health options. The trend, for now, in the United States increases enrollment in managed care plans. Although this is occurring at a rapid pace, managed care will probably not be the final solution to provision of medical care in the United States. PMID- 9347447 TI - Needs assessment at a practice level; using routine data in a meaningful way. AB - BACKGROUND: General practitioners and primary health care teams are asked to take an increasing role in assessing needs and priorities for the people they serve. We describe a model, using routinely available data, to provide health and social information to all general practices in an English Health Authority (South & West Devon, population 586,000, containing 103 general practices) to inform practice based needs assessment. METHOD: Practice-coded hospital activity information was used where available, otherwise the FHSA population register was used to assign spatially referenced data (OPCS birth files, OPCS mortality files, cancer registration files and census information) to practices. Additionally, indicative incidences and prevalences were calculated for each practice using age- and sex specific rates derived from national surveys (Survey of Morbidity Statistics from General Practice and the OPCS Health Survey for England). RESULTS: Information was produced for each practice on births, lifestyle, social factors, incidence and prevalence, hospital activity and mortality. Patient numbers rather than rates were presented. General practitioners commented that this approach gave an understanding of the size of health and social problems and fitted with the concept of numbers needed to treat. CONCLUSION: Public health involvement in practice-based needs assessment is essential. Current public health input has mainly been on a selective individual practice basis and is very resource intensive. This approach allows all practices to have immediate access to a wide range of health and social information presented in a way that is easily understood and informs debate on health needs within the practice. PMID- 9347448 TI - How should public health policy be developed? A case study in European public health. AB - BACKGROUND: Article 129 of the Treaty of Rome (as amended at Maastricht) gave new powers to the European Union (EU) institutions to develop and implement public health programmes at EU level. The authors were invited by the cabinet of Commissioner Flynn to assist the Commission by developing new policy proposals in certain specific areas. METHODS: The approach was agreed with officials of the Public Health Unit (now the Public Health and Safety Directorate) in DG V. Working groups of experts were appointed to review policy options in five discrete areas. The experts were resident in 13 of the 15 current EU member states, and were employed in academic departments, in health ministries, in local government, and in non-governmental organizations. Draft reports were presented to an evaluation panel of additional experts, whose comments contributed to the final report to the Commission. RESULTS: The final report consisted of five main chapters, corresponding to the work of the five working groups. The recommendations were grouped into those for which implementation would be short term, medium term or longer term, and these were summarized as a Plan of Action. CONCLUSION: Five criteria for good public health policy development were satisfied, but the approach used was excessively expensive and time-consuming. Some further lessons for future policy development work have also been recorded. This work provided a fascinating insight into the workings of the EU institutions. PMID- 9347449 TI - A comparison of smokers' and ex-smokers' health-related quality of life. AB - BACKGROUND: The aim of the study was to assess the difference in health status between current smokers and ex-smokers of five years or greater standing. METHODS: A group of current smokers and a group of ex-smokers (of five years or greater standing) in Aberdeen, north-east Scotland, were each sent a postal questionnaire containing SF-36, EuroQol, condition-specific and socio-demographic questions. The subjects were 3000 adults (1500 smokers, 1500 ex-smokers) randomly selected from the records of nine general practices. The main outcome measures were the eight scales within the SF-36 health profile, EuroQol tariff scores and assessment of respiratory symptoms. RESULTS: Smoking cessation leads to an improvement in a range of respiratory symptoms and health-related quality of life. However, in some cases other socio-economic characteristics are better indications of quality of life than smoking status. CONCLUSIONS: Smoking cessation leads to a significant improvement in a range of respiratory symptoms. There appear to be significant differences between smokers' and ex-smokers' perceived quality of life. However, these differences are relatively small and in the majority of cases are better explained by variation in age, housing and economic status. When promoting smoking cessation to patients it is possible to highlight expected improvements in respiratory symptoms, impact on global quality of life and longer-term disease effects. PMID- 9347450 TI - Regional variation in intervention rates: what are the implications for patient selection? AB - BACKGROUND: Whereas geographical variations in intervention rates are well recognized, little is known about their implications for patient selection. This study looks at how the relative probability of being treated in different regions within England vary with a person's need for treatment, and whether higher intervention rates are associated with a greater probability of treatment at all levels of need or confined to only certain levels. METHODS: The method was modelling of retrospective data from population surveys, patient cohort studies and population intervention rates. Two southern regions (SW Thames and Wessex) and two northern regions (Northern and Mersey) were compared. Subjects were men aged 55 years and above in the population with urinary symptoms suggestive of benign prostatic hyperplasia and men undergoing surgical treatment. The ratio of probability of surgery in the southern regions to that in the northern regions by level of symptom severity was determined. RESULTS: The rate of surgery in the southern regions was 26.5 per cent higher than in the north. A higher proportion of patients in the north had severe symptoms before surgery (58 per cent vs 52 per cent; p = 0.002). The probabilities of being operated on in a given year varied by symptom severity in both the north and the south. The probability was higher in the south at all levels of symptom severity: none/mild (ratio = 1.44; p > 0.01), low-moderate (ratio = 1.35; p = 0.003), high-moderate (ratio = 1.53; p < 0.0001), and severe (ratio = 1.15; p > 0.01). On testing the sensitivity of the key assumptions by assuming a more severe distribution of symptoms in the south, the differences at none/mild and low-moderate symptom levels were enhanced but differences at high-moderate and severe symptom levels were reversed. CONCLUSIONS: As few men with mild symptoms qualify for surgery and most men with severe symptoms are operated on, any difference in patient selection between high and low rate regions is inevitably confined to the intermediate group of men with moderate symptoms. Surgeons appear to be rationing their resources in a sensible way, though perhaps not as stringently as could be achieved. PMID- 9347451 TI - A descriptive study of the self-perceived needs of carers for dependants with a range of long-term problems. AB - BACKGROUND: This study aimed to identify qualitatively the need for health and social care of carers looking after dependants from different patient groups in a geographically defined area--Fife, Scotland. It was the first stage of a systematic process designed to assess and meet carers' needs. METHOD: Subjects for the study were unpaid (or 'informal') carers looking after dependants who were known to statutory or voluntary services. A series of 14 focus group discussions with carers of dependants from seven different patient groups took place. The main outcome measure was the qualitative descriptions of carers' self reported health and social needs. RESULTS: Needs 'common' to carers across all care groups were identified. These related to the need for: information (diagnostic, prognostic and where to obtain help), improved communication with professionals, relief from stress, respite care, training and practical support. Within each area of 'common' need, carers had specific needs, which related to the particular needs of their dependant. There were also needs that were identified by carers from one or more patient groups. Carers did not necessarily recognize themselves as carers at an early stage in their caring career. Therefore, their dependants' early medical contact with their general practitioner or hospital specialist was seen by carers as a crucial point at which their own needs for information and help could be recognized. Other opportunities for health professionals to help carers related to involving carers in case management, the provision of counselling and training carers to provide care themselves. CONCLUSION: Many of the needs described by carers were of a social nature. However, carers also described needs relating to the health services--health professionals need to be proactive in recognizing carers' health and information needs and are required to recognize carers' contribution to the welfare of their dependant. In Fife, a multi-faceted approach was used to meet these needs. The challenge for the health service is to find a way to do this at a national level. PMID- 9347452 TI - Breast, lung and colorectal cancer incidence and survival in South Thames Region, 1987-1992: the effect of social deprivation. AB - BACKGROUND: This paper describes the relationship between social deprivation and incidence of, and survival from, breast, lung, and colorectal cancers among residents of the South Thames regions. We analysed 23,505 cases of breast cancer, 29,903 cases of lung cancer and 21,905 cases of colorectal cancer, aged 40-99 inclusive at diagnosis and diagnosed between 1 January 1987 and 31 December 1992. METHODS: Using the 1991 Census in conjunction with the Townsend index on social deprivation, we derived proxy indicators of deprivation based on patients' home postal codes. Cumulative relative five-year survival rates (per cent) were calculated for each cancer. We then compared our results with the relevant standardized incidence and mortality ratios by deprivation status. RESULTS: A clear trend was observed in standardized mortality rates across deprivation tenths for the three tumour sites: mortality increased with deprivation. A strong positive correlation was found between deprivation and the incidence of lung cancers (p < 0.0001), but no association was found between deprivation and incidence of breast and colorectal cancers. Significantly lower five-year relative survival rates were found for breast and colorectal cancer patients in the most deprived Townsend tenths. Breast cancer patients resident in the most affluent tenth of enumeration districts had a 70 per cent relative survival ratio compared with 57 per cent in the most deprived tenth. The corresponding figures for colorectal cancer patients were 40 per cent and 32 per cent, respectively. CONCLUSIONS: Survival differences by deprivation status exist in South Thames among patients suffering from breast or colorectal cancers and are not explained by differences in the incidences of these diseases. For lung cancer, incidence and mortality were positively correlated with deprivation, but no socio-economic gradient was found for survival. PMID- 9347453 TI - Sickness absence and 'working through' illness: a comparison of two professional groups. AB - BACKGROUND: Few studies have investigated occupational groups reporting low rates of sickness absence because of an assumption that these rates indicate low morbidity. This is inconsistent with the view that sickness absence, which may be caused by social and psychological rather than medical factors, does not equate with morbidity. This paper investigates rates of sickness absence and factors influencing decisions not to take sick leave among doctors and a comparative professional group. METHODS: A postal survey was sent to 670 general practitioners (GPs), 669 hospital doctors and 400 company 'fee earners'. Qualitative interviews were conducted with 64 doctors reporting an illness lasting one month or more in the last three years. RESULTS: Self-reported health status was similar for both groups but GPs reported higher levels of occupational stress. However, doctors were significantly less likely to report short periods of sick leave in the previous year. Over 80 per cent of all respondents had 'worked through' illness, citing cultural and organizational factors behind their decision not to take sick leave. Barriers to sick leave among doctors included the difficulty of arranging cover and attitudes to their own health. CONCLUSIONS: Considerable emphasis has been given to the role of social factors in contributing to rates of sickness absence. These may also contribute to the decision not to take sick leave, resulting in possible inappropriate non-use. Measures to encourage and enable doctors to take sick leave might improve the management of their own health. PMID- 9347454 TI - Managing care. AB - The terms 'managed care' and 'disease management' are gaining common usage in the health service but their meaning is not widely understood. Managed care is a generic term describing any health care system that integrates the financing and delivery of medical care. Its growth in the United States has been driven by pressure to control costs, and there is circumstantial evidence that costs are slowing as a result of better management of resources. However, it is not clear how much of this is due to managed care, the selection of more favourable enrollees to health plans or other factors. Research evidence is limited, and that available is constrained by the rapidly changing nature of managed care. In the United States a bewildering variety of managed care arrangements have emerged, although several common characteristics can be identified: limited choice of physician providers; controlled access to secondary care; selective contracting; financial incentives; quality management; and utilization management. All are present in the National Health Service (NHS), which exemplifies a nationalized managed care system. Disease management is an extension of managed care that takes a global approach to patient care by attempting to co-ordinate resources across the entire health care delivery system throughout the life cycle of the disease. This is poorly developed in the NHS, so that the attention of commercial organizations has been attracted. However, concern has been expressed about the implications of commercial involvement: the fragmentation of general medical services; effect of for-profit status; and use of patient-based data. Recent policy developments could allow disease management to develop within the NHS. PMID- 9347455 TI - The application of evidence-based priority setting in a District Health Authority. AB - BACKGROUND: Despite the current drive towards evidence-based medicine, it is not clear how commissioners actually use research findings in making decisions on priority setting. This study describes an attempt to directly introduce evidence on the effectiveness of interventions into the annual priority setting process of a District Health Authority (DHA). METHODS: Literature searches were undertaken on proposals for Health Authority funding. The identified literature was then critically appraised, and relevant information was used by members of the Department of Public Health to score the individual bids in terms of health gain. These scores were then fed into the priority setting process. RESULTS: A total of 144 proposals for funding were submitted. For 6.2 per cent of proposals there was strong evidence to support the intervention, for 21.2 per cent there was fair evidence in support, and for 38.1 per cent there was poor evidence. A search was not possible for 16.8 per cent of the proposals. There was a moderate correlation between the strength of evidence for the effectiveness of the proposal and initial scoring of the proposal for health gain (r = 0.41, p < 0.001). At the end of the priority setting process there was no correlation between strength of evidence and priority ranking (r = 0.01, p = 0.97). CONCLUSIONS: It is feasible, but difficult, to use information resources and critical assessment of research evidence as part of the priority setting process of a DHA. The research evidence did appear to influence the initial assessment of proposals. However, the strength of the research evidence was not associated with the priority choices made by the DHA in its purchasing plan. PMID- 9347456 TI - Control of infection: a survey of general medical practices. AB - BACKGROUND: The aims of the study were (1) to assess current infection control practice within general medical practices and establish a base line; (2) to identify potential infection control problems; (3) to assess the need for local infection control guidelines or standards related to general medical practice; (4) to assess the need for educational provision. METHODS: A survey was carried out, using questionnaire and structured interviews, of all general practices (92) within a Health Board area with a patient population of 561,300. RESULTS: Forty two (46 per cent) practices participated, serving 67 per cent of the patient population. Only three (7 per cent) practices had written infection control policies and only six (14 per cent) provided training on the subject. Thirty (71 per cent) practices had autoclaves; however, performance monitoring was poor. The majority of high-risk instruments were adequately decontaminated; of the medium risk instruments, the auriscope speculum was the item most frequently inadequately treated [36 practices (88 per cent)]. Deficiencies were identified in treatment of blood spillage, and protective clothing provision was variable. The majority, 40 (95 per cent) practices, had systems to deal with clinical waste; however, only two (5 per cent) reported use of BS7320 sharps containers on domiciliary visits. Despite the recognized dangers, 23 (55 per cent) practices resheathed needles and only six (14 per cent) had first aid guidance for needlestick injuries. Only eight (19 per cent) practices knew and recorded staff immunity to hepatitis B following vaccination. CONCLUSIONS: Some deficiencies in infection control practice were identified and the need for policy guidance and staff training was highlighted. PMID- 9347457 TI - Screening immigrants for tuberculosis in Newcastle upon Tyne. AB - BACKGROUND: Successive national guidelines on the control of tuberculosis in the United Kingdom have included recommendations for screening immigrants coming from countries with a high prevalence of tuberculosis. As there has been only one other study on the process and outcome of screening immigrants at a district level the aim of this study was to assess the contribution of screening immigrants to the control of tuberculosis in Newcastle. METHODS: The Port of Arrival (POA) forms were used to identify all new immigrants for screening in Newcastle during 1993. The Family Health Services Authority (FHSA) register was used to identify additional new immigrants from the Indian sub-continent. For all new immigrants identified by the POA forms and FHSA register, hospital and practice records were reviewed for evidence of screening and its outcome up to the end of 1994. RESULTS: There were 252 POA forms in 1993, 100 of which were for immigrants from the Indian sub-continent. This represents less than a third of all new immigrants from the Indian sub-continent. Of all immigrants identified by POA forms, 99 (39 per cent) had been screened. This resulted in the detection of one active case of tuberculosis. CONCLUSIONS: The POA system alone is inadequate for identifying immigrants for screening. The published evidence on screening immigrants for tuberculosis suggests that the system does not work well and the yield of new cases is low. To assess the effectiveness of screening immigrants a national audit is needed. PMID- 9347458 TI - Prevalence of Q fever in a rural practice. AB - BACKGROUND: Q fever is a world-wide condition caused by the rickettsia Coxiella burnetii. It appears more prevalent in agrarian communities and may have serious sequelae. METHODS: A descriptive, cross-sectional, observational study using a randomly selected group of the adult working practice population in a rural practice in West Wales was devised. An immunofluorescence test, which identified past infection, was used to look for associations between C. burnetii seropositivity and farm-related or social activities, and to compare the findings with those of other studies. An attempt was made to establish a clinical profile for the illness Q fever. RESULTS: Twenty-one subjects were found to be seropositive to C. burnetii. No definite consistent clinical features were identified. Farming was undoubtedly a risk factor for the disease, maybe with other related factors also important. There was a possibility that alcohol had a protective effect. No sinister sequelae were described. CONCLUSIONS: Q fever occurs more frequently in farmers than in non-farmers, but was less common than previously thought. Is Q fever accurately described in medical textbooks? A case is made for a more co-operative approach between primary carers and epidemiologists in the study of illnesses in populations. PMID- 9347459 TI - A survey of knowledge and use of folic acid among women of child-bearing age in Dublin. AB - BACKGROUND: The Medical Research Council vitamin trial highlighted the importance of folic acid in the prevention of neural tube defects. Since 1993, the Irish Department of Health has recommended periconceptional folic acid supplements. The objective of this study was to document the knowledge and behaviour of women in child-bearing years to periconceptional folic acid. METHODS: A cross-sectional community-based survey was conducted in Dublin using an interviewer administered questionnaire. RESULTS: A total of 335 women took part in the study, a response rate of 84 per cent. Approximately two-thirds (213/ 335, 63.6 per cent) had heard of folic acid. Knowledge was significantly associated with higher social class and higher education (p < 0.05). Few (18/335, 5.4 per cent), had been advised to take folic acid before pregnancy. Only 9/335 (2.7 per cent) of the women in the study were currently taking folic acid supplements. Three-quarters (75.9 per cent) of the group would be willing to take periconceptional folic acid supplements if they believed it would reduce the risk of malformations. The majority (77.4 per cent) would prefer to take folic acid in tablet form rather than have it added to food. CONCLUSIONS: This study shows that few women in child bearing years in Dublin have been advised on folic acid, and very few are taking supplements. However, if advised appropriately the majority would be willing to take periconceptional folic acid in tablet form. PMID- 9347460 TI - Framework of epidemiological principles underlying chemical incidents surveillance plans and training implications for public health practitioners. AB - The Department of Health requires District Health Authorities to have plans ready for dealing with chemical incidents to protect public health. The three aspects of a chemical incident are environmental, medical toxicology, and public health advice and information. Public health surveillance plans require generation of a causal hypothesis and assessment of risk. Training provision should emphasize the need for application of epidemiological precepts in drawing up such surveillance plans. The epidemiological principles are systematically outlined with emphasis on their significance. PMID- 9347461 TI - Partnership changes in English general practice from 1990 to 1994. AB - BACKGROUND: The objective of this study was to quantify the rate of partnership change among general practitioners (GPs) in the National Health Service (NHS) in England from 1990 to 1994. METHODS: Time series data on English GPs were analysed on 1 October for the years 1990-1994. The main outcome measures include: (1) proportion of GPs practising in an unchanged partnership from 1 October 1990 to 1 October 1994; (2) proportion of partnerships that were unchanged over the study period; (3) the average yearly rate of partnership changes for England and per Family Health Service Authority (FHSA), calculated using both the individual GP and the practice as the unit of analysis. RESULTS: A total of 6532 (27.1 per cent) of the 24,107 unrestricted GPs practising full time on 1 October 1990 were still practising in the identical partnership on 1 October 1994; 3539 (35.7 per cent) of the 9918 practices in England were unchanged over the same period. The average yearly partnership change rate for all England was 23.1 per cent when calculated using the individual GP as the unit of analysis, and 23.4 per cent when calculated using the practice as the unit of analysis. There is threefold variation found in the average yearly partnership change rate by FHSA, with similar rank ordering of health authorities when using either the individual GP or practice as unit of analysis. CONCLUSIONS: Changes in partnerships are commonplace. The possible influence of such changes on primary care in the NHS should be further investigated. PMID- 9347463 TI - Quarterly Communicable Disease Review January to March 1997. From the PHLS Communicable Disease Surveillance Unit. PMID- 9347462 TI - Specialist outreach clinics in Sheffield: a faster tier of out-patient provision for the patients of fundholding GPs? AB - BACKGROUND: Little is known about the activity of the many new specialist outreach clinics in fundholding general practices that have emerged since the introduction of fundholding in 1991, though it has been claimed that specialist outreach clinics have shortened waiting times for fundholders' patients. This study describes the activity of specialist outreach clinics in fundholding practices in Sheffield, focusing on comparative waiting times between fundholding and non-fundholding practices. METHODS: A descriptive study was carried out using routine out-patient activity data and a listing of outreach clinics obtained from fundholding practices. RESULTS: Thirty-seven specialist outreach clinics were established in fundholding practices by November 1994; 23 in surgical specialties. In 1994-1995, for gynaecology, orthopaedics and general surgery, the leading outreach specialties, 22.5 per cent of fundholders' first attendances were in outreach clinics. In those three specialties, 87.0 per cent of patients in specialist outreach clinics in fundholding practices vs 67.1 per cent in hospital clinics were routine appointments, and 17.4 per cent vs 9.4 per cent, respectively, were added to an in-patient waiting list. The proportion of first attendees seen in less than three months was 97.0 per cent in specialist outreach clinics in fundholding practices vs 88.1 per cent in hospital clinics; 90.4 per cent for the patients of fundholders who had outreach clinics vs 85.2 per cent for fundholders who did not; 88.1 per cent for all fundholders' patients vs 88.6 per cent for non-fundholders' patients. CONCLUSIONS: The new specialist outreach service in fundholding practices in Sheffield is largely for surgical patients classified as routine patients. Although patients were seen more quickly in specialist outreach clinics, no overall inequality of waiting times between fundholding and non-fundholding practices was shown. PMID- 9347464 TI - Letter from Zambia 'Naught for your comfort'. PMID- 9347465 TI - 'Syndromes' and reasons not to do a project. PMID- 9347466 TI - NHS performance guides: raising the standard--indirectly? PMID- 9347467 TI - Rheumatic fever. PMID- 9347468 TI - A noxious trio: nausea, gastric dysrhythmias and vasopressin. PMID- 9347469 TI - Neuropeptides in idiopathic chronic constipation (slow transit constipation). AB - Tissue specimens from the large bowel of 18 patients with long-standing slow transit constipation were investigated to determine the distribution and density of several neuropeptides and amines in the enteric nerve system, and also of endocrine cells in comparison to normal individuals. CGRP (calcitonin gene related peptide), galanin, glucagon, GRP (gastrin-releasing peptide), metenkephalin, motilin, neuropeptide Y (NPY), PACAP, peptide YY (PYY), serotonin, somatostatin, substance P and VIP were studied by immunohistochemistry. Tissue concentrations of VIP, substance P and galanin were also measured by radioimmunoassay. Significantly increased VIP, SP and galanin contents were found in specimens from the ascending colon. Levels of VIP and galanin were also increased in the transverse colon. Immunohistochemistry revealed only marginal changes with an increased density of PACAP nerve fibres in the smooth muscle and of VIP and PACAP nerves in the myenteric plexus of the transverse colon. In the descending colon substance P and NPY immunoreactivity were also increased in the myenteric plexus while the density of VIP nerve fibres was reduced in the mucosa/submucosa. The frequency of PYY-containing cells and the 5-HT-containing cells in the ascending colon was significantly increased in the constipated patients. PMID- 9347470 TI - The effect of intravenous vasopressin on gastric myoelectrical activity in human subjects. AB - Vasopressin's role in the sensation of nausea is incompletely understood. In this study, our goals were to investigate whether high intravenous vasopressin levels in normal subjects would induce nausea and vomiting and to determine the electrogastrographic (EGG) pattern which would develop at these concentrations. METHODS: EGG recordings were made on five fasting healthy subjects (three females, mean age: 27 years). Vasopressin was infused (0.15 or 0.3 U kg-1 h-1) for 1 h after a 30-min baseline recording. Serum vasopressin levels were measured every 15 min. Symptoms of nausea, cramping, retching, vomiting and bloating were graded from 0 to 5 (0 = none, 5 = most severe). Normal saline at the same rate was then infused for 1 h, with recording of symptoms and measuring blood levels of vasopressin as done previously. RESULTS: EGG data showed a 43% reduction in the percentage of normal slow waves (96-53%) at a vasopressin rate of 0.3 U kg-1 h-1. A 29% reduction (88-59%) occurred at 0.15 U kg-1 h-1. The EGG dominant frequency decreased by 0.8 cpm (3.07-2.25) for the high dose, while only 0.2 cpm reduction (2.9-2.7) occurred at the lower dose. Bradygastria (< 2.4 cpm) rather than tachygastria (> 3.7 cpm) was the predominant abnormality with the high dose. Symptoms of nausea correlated with the infusion of vasopressin and significantly increased with the higher dose. CONCLUSIONS: (i) At supraphysiological vasopressin levels, nausea was present in 80% of subjects but there was no retching or vomiting, (ii) bradygastria was the predominant dysrhythmia at these high vasopressin concentrations, (iii) increasing vasopressin levels correlated symptomatically with increases in nausea. PMID- 9347471 TI - Measurement of gastric emptying by intragastric gamma scintigraphy. AB - Gastric emptying is usually measured in animals and humans by dilution/sampling or external scintigraphy. These methods are either time consuming or require expensive equipment. The capacity of a miniature gamma counter positioned in the stomach to measure emptying of liquid and solid meals was evaluated. In eight conscious pigs fitted with gastric and duodenal cannulae, gastric emptying of saline (500 mL), dextrose (20%, 500 mL), porridge (300 g) and scrambled eggs (300 g), all labelled with 3.5 MBq 99mTC, was evaluated. When positioned in the antrum the probe was unable to quantify gastric emptying. In contrast, measurements of the fractional emptying of saline over 4-min periods by the probe positioned in the corpus and quantification of radioactivity in the duodenal effluent correlated closely (r = 0.88, P < 0.05). Gastric emptying (50% emptying time) of saline and both solid meals measured by the probe was not significantly different from quantification of the duodenal effluent volume. No difference was observed also for the dextrose meal but only while gastric acid secretion was suppressed by omeprazole. We conclude that an intragastric gamma counter permits measurement of gastric emptying of homogeneous meals provided meal stimulation of gastric secretion was not extensive. This was possible probably by monitoring emptying from the proximal stomach. PMID- 9347472 TI - Pyloric motor response to central and peripheral nitric oxide in the ferret. AB - This study has investigated the relative importance of central nervous and peripheral nitroxidergic mechanisms in the control of pyloric motility. In 10 urethane-anaesthetized ferrets, drugs were administered directly to the CNS via a 0.5-mm-diameter cannula inserted into the 4th ventricle, approximately at the obex. Drugs were also given directly to the upper GI tract by close intra arterial (i.a.) injection at the coeliac axis. Antropyloroduodenal pressures were recorded with a five-channel sleeve/sidehole micromanometric assembly (1.35 x 1.75 mm o.d.), which was introduced via the duodenum. Pyloric motility was stimulated throughout the main part of each study with a continuous i.v. infusion of CCK-8 (30 pmol min-1). This infusion produced an immediate and sustained increase in tonic and phasic pyloric activity, and sustained abolition of antral pressure waves. CCK-8 also induced a duodenal motor response, but this was short lived (11.4 +/- 7.9 min). Coeliac axis injection of the NO donor S-nitroso-N acetyl-penicillamine (SNAP) decreased phasic pyloric activity (from 330 +/- 35 to 148 +/- 21 mmHg min-1 after SNAP 5 micrograms, P < 0.01). By comparison central SNAP administration over the same dose range had no effect on CCK-stimulated pyloric motlity. Inhibition of endogenous NO synthase with L-Nitro Arginine Methyl Ester (L-NAME, 100 mg kg-1 close i.a.) caused a marked increase of phase pyloric motor activity from 349 +/- 59 to 1044 +/- 140 mmHg min-1 (P < 0.01). In addition, SNAP caused marked stimulation of pyloric tone from 2.6 +/- 0.5 to 13.1 +/- 2.8 mmHg (P < 0.01). Central nervous administration of L-NAME caused modest enhancement of phasic pyloric activity (248 +/- 31 to 283 +/- 32 mmHg min-1 P < 0.05) and pyloric tone (2.6 +/- 0.5 to 3.7 +/- 0.7 mmHg, P < 0.05). Our data indicate that motor activity of the ferret pylorus is potently modulated by NO released within the upper gut. Additionally, there is potential for modulation of pyloric motility by central nervous system production of NO. PMID- 9347473 TI - Comparison of lower oesophageal sphincter pressure measurement using circumferential vs unidirectional transducers. AB - Average values from unidirectional (UND) transducers have traditionally been used in standard manometric evaluation of lower oesophageal sphincter pressures (LOSPs). A single circumferential (CMF) transducer has recently become popular. Our aim was to compare LOSP measurements made using the two techniques. Ten healthy volunteers were intubated with a Konigsberg manometry catheter containing five solid-state transducers (four unidirectional at 90 degrees angles radially and 2.5-cm intervals and one circumferential, 3 cm distal). LOSP was measured using the station pull-through technique in the supine and upright positions with Synectics Polygram software for computerized analysis. Mean UND pressures were plotted against CMF pressures and a linear regression and correlation analysis was performed. A similar analysis was done for LOS length. A significant difference (P < 0.01) was found between mean (+/- SE) supine and upright LOSPs for both mean UND (20.21 +/- 1.36 vs 15.56 +/- 1.28 mmHg) and CMF (20.41 +/- 1.96 vs 16.17 +/- 1.61 mmHg); however no difference was shown between supine and upright lengths. There was a high degree of correlation between LOSP (r = 0.83) and a weaker correlation (r = 0.65) between LOS lengths when measured by CMF and UND transducers. CONCLUSION: UND and CMF transducers provide identical measurements when evaluating the LOS. PMID- 9347475 TI - Gastric contractions, secretions and injury in cold restraint. AB - The clinical syndrome of stress ulceration has been studied for years using rodent cold restraint stress models, although the pathogenesis of the characteristic focal gastric mucosal lesions produced in these models has been controversial. We used gastric strain gauges to characterize fully the gastric motility effects of a 4-h cold restraint protocol, and we determined the relationship of variations in gastric contents and in gastric contractions to the amount of gastric mucosal injury. Additionally, we examined rat stomachs histologically, and determined the location of focal haemorrhagic mucosal lesions on the mucosal rugae. We found a consistent relationship between force of gastric contractions and gastric mucosal injury, and also a relationship between the initial duration of contractions during restraint and ultimate mucosal injury. Volume, acidity and mucus in the gastric contents were unrelated to mucosal injury. The majority (91%) of the mucosal lesions had some relationship to a rugal fold, with 59% of all lesions at the base of a rugal fold. Thus, the mechanical forces of gastric hypercontractility may contribute to the gastric mucosal injury of rodent cold restraint models. PMID- 9347474 TI - 5-HT activates neural reflexes regulating secretion in the guinea-pig colon. AB - The role of 5-hydroxytryptamine (5-HT) in neural reflexes regulating secretion was examined in muscle-stripped segments of guinea-pig colon set up in modified flux chambers. A 15-microL pulse of 5-HT (100 microM) to the mucosal bath (1.5 mL), which was continuously perfused, evoked an increase in short-circuit current (Isc). The 5-HT-induced increase in Isc was inhibited by tetrodotoxin, N-acetyl-5 hydroxytryptophyl-5-hydroxytryptophan amide (5-HTP-DP), GR82334 and atropine, but not by tropisetron. 5-HTP-DP reduced the response to a 5-HT pulse over the concentration range of 1 nM to 1 microM. The Isc response to a 5-HT pulse was unaffected by the cyclooxygenase inhibitor, piroxicam. This contrasted with a reduction in the Isc response to mucosal stroking with a brush by piroxicam. The results suggest that a 5-HT pulse, like mucosal stroking, activates a secretory reflex that includes tachykinin and cholinergic neurons but, unlike mucosal stroking, does not release prostaglandins. PMID- 9347476 TI - Sodium azide induces relaxation of the canine gastric body by activating a guanylate cyclase-dependent pathway. AB - In order to study the inhibitory mechanism by which sodium azide eliminates smooth muscle contraction in vivo and in vitro, gastrointestinal motility was monitored via chronically implanted force transducers in the stomach and duodenum of conscious dogs. Circular smooth muscle strips with myenteric plexus from the canine gastric body were used for in vitro measurement of isometric tension. In conscious dogs, sodium azide (50 micrograms kg-1, i.v.) abolished both the spontaneously occurring phase III contractions and postprandial motility. Exogenous motilin (100 ng kg-1)- and bethanechol (50 micrograms kg-1)-induced contractions were also abolished by sodium azide. In vitro, sodium azide and electrical field stimulation (EFS) caused a concentration- or frequency-dependent nonadrenergic noncholinergic relaxation in the gastric body strips. The relaxation induced by EFS, but not sodium azide, was abolished by tetrodotoxin. NG-nitro-L-arginine and oxyhaemoglobin failed to attenuate the relaxant effect of sodium azide, but strongly inhibited EFS-induced relaxation. Methylene blue inhibited both sodium azide- and EFS-induced relaxation. cGMP concentrations in muscle strips were markedly increased by sodium azide. These findings indicate that sodium azide induces relaxation in the canine gastric body through a direct action on smooth muscle, by activating a guanylate cyclase-dependent pathway; endogenous NO synthesis does not participate in this inhibitory mechanism. PMID- 9347477 TI - Symptoms and gastric functions in dyspepsia--goodbye to gastroparesis or to inadequate studies? PMID- 9347478 TI - A neural basis for the retrieval of conceptual knowledge. AB - Both clinical reports and systematic neuropsychological studies have shown that patients with damage to selected brain sites develop defects in the retrieval of conceptual knowledge for various concrete entities, leading to the hypothesis that the retrieval of knowledge for entities from different conceptual categories depends on partially segregated large-scale neural systems. To test this hypothesis, 116 subjects with focal, unilateral lesions to various sectors of the telencephalon, and 55 matched controls, were studied with a procedure which required the visual recognition of entities from three categories--unique persons, non-unique animals and non-unique tools. Defective recognition of persons was associated with maximal lesion overlap in right temporal polar region; defective recognition of animals was associated with maximal lesion overlap in right mesial occipital/ventral temporal region and also in left mesial occipital region; and defective recognition of tools was associated with maximal lesion overlap in the occipital-temporal-parietal junction of the left hemisphere. The findings support the hypothesis that the normal retrieval of knowledge for concrete entities from different conceptual domains depends on partially segregated neural systems. These sites may operate as catalysts for the retrieval of the multidimensional aspects of knowledge which are necessary and sufficient for the mental representation of a concept of a given entity. PMID- 9347479 TI - Explaining category-related effects in the retrieval of conceptual and lexical knowledge for concrete entities: operationalization and analysis of factors. AB - Category-related effects in the retrieval of conceptual and lexical knowledge for concrete entities have been well documented in lesion studies, and also with functional imaging and electrophysiological approaches. For example, brain damaged subjects may be impaired in the ability to recognize or to name animals but not tools, or the opposite pattern may obtain. One reason for these dissociations is that different patterns of defects tend to be caused by distinct lesion profiles, suggesting a relative tendency for certain neural systems to be involved in category-related knowledge. But we and others have also hypothesized that a variety of traits of concrete entities co-determine category-related dissociations. Such traits ('factors') include homomorphy (similarity of form), familiarity, value to perceiver, manipulability, characteristic motion, characteristic sensory modality of transaction (vision, touch, hearing), and typical age of acquisition. It is our view that the mix of factors relative to different conceptual categories plays a key role in the neuroanatomical distribution of records for those different categories, and is thus behind the systematic correlations between certain retrieval defects and damage to certain neural systems [12, 52]. In this study, we operationalized these factors and analyzed their intercorrelations. Stimuli were slides of 215 items from the conceptual categories of animals, fruits/vegetables, tools/utensils, vehicles, and musical instruments. The factors were operationalized on the basis of ratings obtained from 227 normal control subjects and on the basis of computer analyses of the digitized outlines of the stimuli. Principal components analysis revealed that 81% of the variability across items could be accounted for by three components: Component 1 (practically useful, common items): high value to perceiver, tactile mode of transaction, high familiarity, low age of acquisition; Component 2 (homomorphic, non-manipulable items): high homomorphy, low characteristic motion and manipulability; Component 3 (items with characteristic sound): hearing mode of transaction, highly distinctive sounds. In another analysis, we found that the categories of animals versus tools/utensils differed significantly on the factors of homomorphy, familiarity, value, manipulability, characteristic motion, and touch. The factor structure we identified in this study may help explain category-related performance defects in brain-damaged subjects. The results lend support to our proposal that systematic differences in physical characteristics and contextual specification of concrete entities constitute a driving force behind the regionalization of neural systems related to the acquisition and retrieval of conceptual and lexical knowledge. PMID- 9347480 TI - Working memory impairments in traumatic brain injury: evidence from a dual-task paradigm. AB - Although many individuals with traumatic brain injury (TBI) perform well on standard neuropsychological tests, they often exhibit marked functional difficulties. The functions which are impaired seem to be analogous to the role of the central executive system (CES) in Baddeley's [Working Memory, 1986, Oxford University Press, New York] widely accepted model of working memory. The purpose of this study was to investigate CES function in individuals with TBI with a dual task paradigm. We studied 25 non-demented persons who were at various stages in their recovery from severe TBI and compared their performance on a dual-task paradigm to a group of age-matched controls. Our dual-task paradigm measured performance on a simple visual reaction time task both alone (baseline) and during concurrent tasks of articulation or digit span. Subjects were also assessed with other neuropsychological tests of executive function. TBI patients had slower reaction times on the primary task when performed alone (P < 0.05) and greater decrements in performance during dual-task conditions (P < 0.01). They also exhibited significantly greater deficits than control subjects on other measures of executive function. Although correlations between dual-task performance and other executive measures were quite low, principle components analysis suggested that a common factor does exist between these measures. These findings support the conclusion that TBI patients have a working memory impairment that is due to dysfunction of the CES and which may be related to executive function deficits as measured by standard neuropsychological testing. PMID- 9347481 TI - Transcranial magnetic stimulation reveals a hemispheric asymmetry correlate of intermanual differences in motor performance. AB - Hemispheric asymmetries in the threshold for eliciting motor evoked potentials (MEPs) with transcranial magnetic stimulation (TMS) are associated with hand preference. We posited that hemispheric asymmetries in TMS thresholds may be strongly correlated with some hand-differences in motor performance. MEPs result from the activation of neuronal networks targeting large cortical motoneurons. Thus, MEP thresholds might reflect physiological features of the corticospinal motor system. Considering the role of corticospinal pathways in the control of independent finger movement, we hypothesized that MEP thresholds would better predict speed and dexterity than strength. In 30 right-handers and 30 left handers, we correlated right and left hand-differences in the threshold for eliciting MEPs with hand-differences in the performance of three manual tasks: finger-tapping speed, pegboard dexterity, and grip strength. Correlations of hand differences in TMS thresholds with hand-differences in performance indicated that a lower TMS threshold for one hand is strongly associated with greater ability with that hand. The correlations of hand-differences in TMS thresholds with hand differences in finger-tapping and pegboard dexterity were significantly larger than the correlation of hand-differences in TMS thresholds with hand-differences in grip strength. Our results indicate that hemispheric asymmetries in MEP thresholds may have functional significance related to basic parameters of movement. These results are consistent with the critical role of the corticospinal motor system in the control of independent finger movement. Furthermore, they imply that asymmetry in the corticospinal motor system may be an important substrate for asymmetries in hand preference and performance. PMID- 9347482 TI - Non-associative lexical priming is impaired in Alzheimer's disease. AB - A word-fragment completion task was used to assess long-term, non-associative lexical priming in patients with probable Alzheimer's disease (AD) and age- and education-matched elderly normal control (NC) subjects. Despite equivalent baseline performance, the AD patients exhibited less facilitation in their ability to complete word fragments from having previously read the intact words than did the NC subjects. The AD patients were also impaired relative to NC subjects on an explicit recognition memory task, but there was no relationship between explicit memory performance and priming for either group. These results are consistent with previous demonstrations of impaired semantically-based priming in patients with AD and extend the domain of their impairment to priming that is predominantly based on lexical activation. PMID- 9347483 TI - Dissociating working memory from task difficulty in human prefrontal cortex. AB - A functional magnetic resonance imaging (fMRI) study was conducted to determine whether prefrontal cortex (PFC) increases activity in working memory (WM) tasks as a specific result of the demands placed on WM, or to other processes affected by the greater difficulty of such tasks. Increased activity in dorsolateral PFC (DLPFC) was observed during task conditions that placed demands on active maintenance (long retention interval) relative to control conditions matched for difficulty. Furthermore, the activity was sustained over the entire retention interval and did not increase when task difficulty was manipulated independently of WM requirements. This contrasted with the transient increases in activity observed in the anterior cingulate, and other regions of frontal cortex, in response to increased task difficulty but not WM demands. Thus, this study established a double-dissociation between regions responsive to WM versus task difficulty, indicating a specific involvement of DLPFC and related structures in WM function. PMID- 9347484 TI - How far does the brain lateralize?: an unbiased method for determining the optimum degree of hemispheric specialization. AB - The relationship between measures (of size or function) on one side of the brain, in relation to the difference between the two sides on that measure, are important components of theories of hemispheric asymmetry. For example, it has been concluded that increasing lateralization (e.g., of hand skill or planum temporale area) occurs at the expense of the non-dominant hemisphere. Here it is demonstrated that such relationships could merely be a necessary consequence of relating components of a laterality index to the index (L - R)/(L + R) itself, or indeed to L - R. An alternative approach (using random data to exemplify the null hypothesis) is presented together with an application to data on hand skill from 12,782 11-year-olds in a cohort study. This demonstrates a symmetry hitherto undocumented of maximal hand skill in left and right hands in left- and right hand writers respectively, the point of the maximum falling short of the population mean for relative hand skill in either case. If degrees of laterality are what is genetically determined, this suggests that selection is present for a function (perhaps language) associated with a greater magnitude of lateralization than is represented by hand skill. PMID- 9347485 TI - A case of callosal agenesis with strong anatomical and functional asymmetries. AB - A 30-year-old right-handed man (W.D.) with total callosal agenesis was examined neuropsychologically and with magnetic resonance imaging. Basic neuropsychological testing revealed normal intelligence and average attentional capabilities. Anatomically, W.D. shows strong leftward perisylvian asymmetry both for the planum temporale and planum parietale, an unusual pattern not found in our database of more than 200 brains of young and healthy individuals. Functionally, W.D. has strong right hand superiority for hand skill and tactile object recognition, indicating unusual left hemisphere dominance for both functions. Our observations could support the hypothesis that callosal connectivity and hemispheric asymmetry may be inversely related. PMID- 9347487 TI - Physiological and neuropsychological correlates of hostility. AB - This experiment tested two hypotheses linking right cerebral arousal to hostility and physiological arousal. A replication of previous research supporting heightened physiological (systolic blood pressure, diastolic blood pressure and heart rate) reactivity among high-hostility subjects was partially successful. Hemispheric lateralization of cerebral activity in response to stress was also measured. Low- and high-hostility subjects were identified using the Cook-Medley Hostility Scale (CMHS). Physiological measures (systolic blood pressure, diastolic blood pressure and heart rate) were recorded and dichotic listening procedures were administered before and after administration of the cold-pressor paradigm. The primary finding of this research was greater right cerebral activation to stress among high-hostility subjects, as indicated by their enhanced ability to identify syllables presented to the left ear. Data further supported previous findings of heightened physiological reactivity to stress among high-hostility subjects and suggest a positive relationship between right cerebral activity and cardiovascular arousal. PMID- 9347486 TI - Differential neural response to positive and negative feedback in planning and guessing tasks. AB - The neural mechanisms by which emotional and cognitive processing interact are unknown. Evidence from animal studies and neurological patients suggests that regions of the ventral striatum and orbitofrontal cortex, together with limbic structures such as the amygdala, are critical to such interactions. We used positron emission tomography to study the neural systems engaged by processing performance feedback under two conditions involving either a complex cognitive or a matched guessing task. The main activations associated with the processing of performance feedback under different task conditions involved foci in the medial caudate nucleus and the ventromedial orbitofrontal cortex. A differential modulation of these activations as a function of task type was observed. In particular the orbitofrontal activation associated with the presence of feedback was only seen in the guessing task. These data suggest that the ventral striatum and orbitofrontal cortex are involved in processing of feedback information, findings consistent with animal and neurological studies. We propose that differential activation associated with guessing compared to planning suggests enhanced neural processing of feedback when the outcome of a task is uncontrollable or when information must be assimilated across a number of trials to assess performance. PMID- 9347488 TI - The time course of spatial and object learning in Parkinson's disease. AB - Parkinson's disease (PD) is characterized by spatial memory dysfunction, but the selectivity of the deficit remains unclear. We addressed this issue by comparing performance on spatial and object variants of a conditional associative learning task, and by analysing the data with time series analytical techniques. The 11 PD subjects and 15 normal control subjects learned stimulus-stimulus pairings through trial-and-error learning. PD subjects were selectively impaired on the spatial condition: they required more trials to achieve criterion, learned at a slower rate and displayed a working memory deficit. The groups did not differ in the object condition. These results suggest a distinction between material specific spatial and object visual memory systems. Further, they indicate that spatial learning and memory are selectively impaired in early PD, suggesting that interactions between the basal ganglia and prefrontal cortex are important for the mediation of high-level cognition. PMID- 9347489 TI - Isn't it about time we changed our perception of diagnostic radiation risks? PMID- 9347490 TI - Nitric oxide production in the lesions of temporomandibular disorders and gender differences in nitric oxide production. PMID- 9347491 TI - Abolish dental insurance? PMID- 9347492 TI - A rapidly growing pigmented plaque. PMID- 9347493 TI - The Millard rotation-advancement lip repair using accurate measurements. AB - For many years a wide variety of surgical techniques for closure of unilateral cleft lip has been used. Still many surgeons prefer the Millard rotation advancement lip repair because the surgical scar is masked in the filtrum crest and the nostril floor, and it improves the relationship of the alar base of the cleft side, producing harmonious symmetry of the nostril and the nostril sill. In addition, it uses and preserves the lip anatomy, returning lip tissue into its normal position, minimizing the amount of tissue that is discarded, and reconstructing the orbicular oris muscle. One of the major disadvantages of this procedure is the lack of accurate measurements. This article demonstrates that rotation-advancement lip repair can be performed with accurate measurements as in the Tennison-Randall lip repair and tends to invalidate the widely held belief that rotation-advancement lip repair is a "cut as you go" technique. PMID- 9347495 TI - Lingual nerve paresthesia following third molar surgery: a retrospective clinical study. AB - Lingual nerve anesthesia, paresthesia, and dysesthesia are possible side effects of third molar extraction. These unwanted complications are frequently disturbing to both the patient and practitioner. The incidence of lingual nerve damage following third molar surgery is more frequent than once thought. Six hundred questionnaires were sent to randomly selected Fellows of the American Association of Oral and Maxillofacial Surgeons in 50 states to determine the parameters surrounding this phenomenon. Of the 452 respondents, 76.05% reported having had patients with lingual anesthesia, dysesthesia, or paresthesia. Of all the reported cases, 18.64% of the cases failed to resolve. Of the reported cases, only three underwent surgical intervention. Because many cases of lingual nerve dysfunction do not resolve, it is important to inform patients that microsurgical nerve repair techniques are available as a modality of treatment following diagnosis. It has also been recommended that if the paresthesia does not resolve within 10 to 12 weeks, then management options including microsurgical nerve reconstruction within a short period of time should be discussed as a plan with the patient. PMID- 9347494 TI - Management of solid ameloblastoma of the jaws with liquid nitrogen spray cryosurgery. AB - OBJECTIVE: This study evaluated the results of the use of curettage followed by liquid nitrogen spray cryosurgery in a number of solid or multicystic ameloblastomas of the jaws and the postoperative complications related to this treatment modality. STUDY DESIGN: Thirty-six patients with solid ameloblastoma of the jaws were treated with curettage followed by cryosurgery. The cryotherapy consisted of hand instrumented curettage of the bone lesion followed by three freezing cycles, of 1 minute each, of the remaining bone cavity with liquid nitrogen spray. Postoperative complications were evaluated clinically and radiographically. RESULTS: Local recurrence occurred in 11 (30.6%) patients. Excepting local recurrence, postoperative complications were frequent but not severe: wound dehiscence (5.5%), paraesthesia (5.5%), infection (5.5%), and pathologic fracture (11.1%). CONCLUSION: Management of solid or multicystic ameloblastomas of the jaws with curettage followed by cryosurgery may decrease the local recurrence rate and also to reduce the initial indication of resection with continuity defect. PMID- 9347496 TI - Oral pemphigus vulgaris: a review of the literature and a report on the management of 12 cases. AB - Twelve cases of oral pemphigus vulgaris are described to illustrate the long-term behavior of the disease and the treatment challenges it presents to the oral medicine practitioner. In addition, we review the literature on oral pemphigus vulgaris with respect to clinical history, signs and symptoms, management, and treatment outcome. Pemphigus vulgaris is a chronic vesiculobullous disease with a potentially fatal outcome. Mortality from pemphigus vulgaris before the development of effective therapies was as high as 90%. Today, with treatment, it is closer to 10%. Involvement of the oral mucosa is common and in most cases precede skin lesions; in our patients, the oral lesions preceded the development of extraoral disease in 75% of cases. Pemphigus vulgaris was more frequent among women (9:3), and there was a tendency for the severity and frequency of disease to decrease with time. PMID- 9347497 TI - Sustained relief of oral aphthous ulcer pain from topical diclofenac in hyaluronan: a randomized, double-blind clinical trial. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that topically applied 3% diclofenac in 2.5% hyaluronan reduces aphthous ulcer pain. STUDY DESIGN: A randomized, double-blind, single dose study of 60 healthy adults with aphthous ulcers in three treatment groups--3% diclofenac in 2.5% hyaluronan, 2.5% hyaluronan, 3% viscous lidocaine--was undertaken. Visual analogue scale pain scores were obtained before and after gel application and hourly, for up to 8 hours after gel application. Statistical analysis was performed with repeated measures ANOVA with square root transformation and Bonferroni correction. RESULTS: A 48% overall reduction in pain (p < 0.01) was observed 10 minutes after gel application; however, no significant difference was found between the three topical agents. A 35% to 52% pain reduction (p < 0.01) was reported 2 to 6 hours after the application of diclofenac in hyaluronan, whereas hyaluronan gel alone and viscous lidocaine failed to produce significant VAS reductions. CONCLUSIONS: A dose of 3% diclofenac in 2.5% hyaluronan is an effective and novel treatment for this common, painful disorder. PMID- 9347498 TI - Thalidomide-induced perioral neuropathy. AB - Thalidomide was administered as a therapeutic agent for chronic graft-versus-host disease after allogeneic peripheral blood stem cell transplantation in a patient with breast cancer. Although side effects of thalidomide have been described earlier, this is the first instance of perioral neuropathy associated with thalidomide treatment. Awareness of this specific side effect may contribute to early diagnosis and appropriate treatment. PMID- 9347500 TI - Oligodontia of the permanent dentition in two sisters with polycystic ovarian syndrome: case reports. AB - Oligodontia or severe hypodontia is a rare developmental dental anomaly commonly associated with syndromes and systemic abnormalities. This report presents two sister, aged 18 and 21, who collectively had 56 congenitally missing permanent teeth. Both patients exhibited pubertal hirsutism, menstrual disturbances, and enlarged ovaries with multicystic lesions defined ultrasonically. These features are consistent with the diagnosis of polycystic ovarian syndrome, a disease not previously linked to hypodontia. A genetic component of this condition is proposed. The significance of this entity is discussed and the importance of early diagnosis and treatment regimens are emphasized. PMID- 9347499 TI - HIV prevalence in dental outpatients in Brazil. AB - A series of dental outpatients in Brazil was anonymously screened for HIV antibodies in whole unstimulated saliva with an immunoglobulin G antibody-capture enzyme-linked immunosorbent assay. Salivary HIV antibodies were detected in 40 patients in the control group who were known to be HIV-seropositive but were not detected in any of a series of 40 known HIV-seronegative patients in the control group, confirming the very high sensitivity and specificity of the immunoglobulin G antibody-capture enzyme-linked immunosorbent assay. Only one patient from 84 consecutive dental outpatients of unknown HIV serostatus who were examined anonymously for HIV by immunoglobulin G antibody-capture enzyme linked immunosorbent assay showed HIV positivity (1.2% of the population). PMID- 9347501 TI - Oral keratinocyte immune responses in HIV-associated candidiasis. AB - INTRODUCTION: Candidiasis is the most commonly encountered opportunistic infection among HIV-positive subjects. The purpose of this study was to assess specific keratinocyte immune parameters in the pseudomembranous and erythematous forms of HIV-associated oral candidiasis. MATERIAL/METHODS: This collaborative study from three centers analyzed 25 HIV-positive and 10 HIV-negative subjects with either pseudomembranous or erythematous candidiasis. Oral biopsy specimens from lesional tissues were procured, and histopathologic features were correlated with immunohistochemical and in situ hybridization investigations for the expression of interleukin 1 alpha, interleukin 8, antimicrobial calprotectin, lymphocyte populations, and Candida antigen. RESULTS: Both pseudomembranous and erythematous candidiasis among HIV-infected subjects showed a mild interface lymphocytic mucositis with the presence of neutrophilic subcorneal abscesses in the latter. Erythematous candidiasis cases that failed to show surface mycelia, did yield positive results for Candida antigens in the parakeratinized layer. The expression of inflammatory chemokines were positive in all groups and calprotectin appeared to serve as a keratinocyte barrier to hyphal penetration. CONCLUSIONS: The erythematous form of candidiasis is often devoid of hyphae yet the presence of Candida antigens in the surface epithelium implicates an immune or allergic process. The intactness of chemokines and antimicrobial calprotectin in keratinocytes may explain why disseminated candidiasis is rarely encountered in HIV-infected patients. PMID- 9347502 TI - Benign neural tumors of the oral cavity: a comparative immunohistochemical study. AB - To determine if immunohistochemistry can be used as adjunct to the diagnosis and classification of oral benign neural tumors, we stained 77 neurally differentiated tumors with a panel of neural-associated antibodies (S-100 protein, CD57, epithelial membrane antigen, factor XIIIa, CD34, CD68, collagen IV). Using standard histologic criteria, we identified 13 schwannomas, 16 neurofibromas, 23 traumatic neuromas, 16 palisaded and encapsulated neuromas, and 9 granular cell tumors from archived oral pathology specimens. Silver stains showed that neurofibromas, traumatic neuromas, and palisaded and encapsulated neuromas consistently contained axon filaments. Although all neural tumors contained S-100-positive cells, schwannomas and palisaded and encapsulated neuromas contained the most. All tumors expressed CD57; traumatic neuromas were stained intensely and the others stained weakly. The consistent epithelial membrane antigen capsular staining of schwannomas and the absence of factor XIIIa positive dendritic/spindle cells helped distinguish these tumors from others. Many CD34-positive cells were found in schwannomas, and few were found in palisaded and encapsulated neuromas. Variable numbers CD68-positive cells were seen in all neural tumor types; some of these cells appeared to be macrophages and mast cells, but many were thought to be Schwann cells expressing this antigen. Collagen IV staining, apparently representing basement membrane, was generally a feature of all benign neural tumors. The immunophenotype of the granular cells of the GCTs was S-100+, CD57+, and collagen IV+ supporting the putative neural origin of these tumors. We conclude that neural origin/differentiation of a connective tissue tumor can be confirmed with stains for S-100 protein, epithelial membrane antigen, CD57, and collagen IV. Staining patterns and intensities associated with the panel of antibodies tested can be useful in tumor classification. PMID- 9347504 TI - Perineural fibrous thickening within the dental pulp in type 1 neurofibromatosis: a case report. AB - A case of type 1 neurofibromatosis is presented that illustrates oral manifestations and their role in the diagnosis of this condition. The oral lesions may be overlooked in the diagnosis of intraoral swellings. This case documents the finding of perineural fibrous thickening within the dental pulp. Such changes may indicate pulpal involvement in neurofibromatosis and the effect of a genetically transmitted disorder upon the pulp. PMID- 9347503 TI - Polymorphous low-grade adenocarcinoma: flow cytometric, p53, and PCNA analysis. AB - Polymorphous low-grade adenocarcinoma of minor salivary glands (terminal duct carcinoma, lobular carcinoma) was first defined more than a decade ago. A 17% recurrence rate and a 9% metastasis rate have been reported. Fifteen formalin fixed, paraffin-embedded archival cases were analyzed. Ploidy and proliferative activity were evaluated with flow cytometric analysis. Demonstration of an abnormal p53 gene product and proliferative cell nuclear antigen analyses were also performed with routine immunohistochemical procedures. The purpose of this investigation was to evaluate these parameters and determine if a correlation existed. Flow cytometry was performed on 10 cases; 3 showed an aneuploid cell line (mean, S-phase diploid tumor cells 5.9%; S-phase aneuploid 26.7%). Products of a mutation of the p53 tumor suppressor gene have been noted to accumulate in salivary gland tumors, both benign and malignant. Qualitative assessment revealed p53 positive staining in 4 of 15 tumors; positive cells comprised 5% to 10% of the tumor. The percentage of tumor cells positive for proliferative cell nuclear antigen staining ranged from 0.5% to 70%. There was no correlation between proliferative activity as determined by proliferative cell nuclear antigen when compared with results of flow cytometric analysis except for one case that exhibited p53 staining, a 26% proliferative cell nuclear antigen fraction, and a distinct aneuploid cell line. PMID- 9347505 TI - Nasolabial cyst: report of a case with extensive apocrine change. AB - The nasolabial cyst is a rare condition of the midline with an uncertain pathogenesis. This case report describes a 44-year-old woman in which the clinicopathologic findings were consistent with nasolabial cyst. On histologic examination, extensive apocrine change was noticed. To our knowledge, this feature has not been mentioned in previous papers. In addition, cases reported during the last decade are summarized. PMID- 9347506 TI - Intra-articular fibroma of tendon sheath in the temporomandibular joint. AB - Fibroma of tendon sheath is an uncommon soft tissue tumor. The first case involving the temporomandibular joint is reported here. The patient presented with chronic clicking, pain, and swelling of the right temporomandibular joint associated with restricted jaw opening. Histologic, immunocytochemical, and ultrastructural features of the intra-articular tumor were identical to fibroma of tendon sheath. The variability of symptoms and the diagnostic problems presented by this tumor are discussed. PMID- 9347508 TI - Removal of intracanal smear by doxycycline in vitro. AB - Cleansing and shaping result in a smear layer on the instrumented canal wall surfaces. The smear layer may inhibit close contact between sealers and dentin, and inhibits diffusion of medicaments. OBJECTIVE: This study assessed the effect of doxycycline hydrochloride (DH) on smear layer on intracanal walls. STUDY DESIGN: Scanning electron microscopy was used to evaluate the remaining smear layer using different concentrations of DH. Single-canal palatal roots of extracted maxillary molars were irrigated with saline-15% EDTA; saline-25 mg/ml DH; saline-50 mg/ml DH; saline-100 mg/ml DH; NaOCl-15% EDTA; NaOCl-25 mg/ml DH; NaOCl-50 mg/ml DH; and NaOCl-100 mg/ml DH. The roots were fractured into halves and the amount of smear layer assessed in the middle and apical third. RESULTS: Doxycycline-HCl of 100 mg/ml was the most effective in removing smear layer. In the saline group, 100 mg/ml of DH was more effective than EDTA. In the hypochlorite group, 50 mg/ml and 100 mg/ml of DH were more effective than EDTA (p < 0.05). CONCLUSION: Doxycycline solution may be an effective irrigant. PMID- 9347507 TI - Laser Doppler flowmetry: a clinical test of pulpal vitality. AB - PROBLEM: A rapid, accurate, noninvasive method of determining pulpal blood flow would be helpful in determining pulpal vitality. OBJECTIVE: The purpose of this study was to determine if laser Doppler flowmetry can measure induced changes in pulpal blood flow. STUDY DESIGN: Two percent lidocaine with epinephrine 1:100,000 was infiltrated into the labial vestibule to anesthetize five anterior teeth in healthy human volunteers. Stents were placed on the teeth to stabilize the laser Doppler probes. Measurements of pulpal blood flow were made along with electrocardiograms to record the cardiac cycle. RESULTS: Laser Doppler flowmetry demonstrated pulpal blood flow and pulse amplitude decreases under test conditions. These decreases were most significant at 10 minutes after the injection of anesthetic with vasoconstrictor. CONCLUSIONS: Laser Doppler flowmetry was able to measure pulpal blood flow and record changes in blood flow that occurred when epinephrine was used with infiltration anesthesia. The pulse width and the mean flow were dramatically affected as was synchronization with the cardiac cycle. PMID- 9347510 TI - Bone height measurements on panoramic radiographs: the effect of shape and position of edentulous mandibles. AB - OBJECTIVE: The aim of this study was to quantify the effect of mandibular angulation, position, and shape of an edentulous mandible on the distortion of its image in panoramic radiographs. STUDY DESIGN: Five edentulous dry mandibles varying in size from small to wide and equipped with metal bars in and on top of the mandible were used. The mandibles were radiographed at nine different positions by tilting the mandible posteriorly around a transversal axis, using an orthopantomograph. RESULTS: The length of the images of the bars on top of the mandible increased significantly by tilting the mandibles from +20 degrees to -20 degrees. The magnification factor of the images of the intrabony bars in the mandible was the largest at 0 degrees and decreased significantly by both decreasing or increasing the inclination. The size of the mandible was not related to the magnification factor. CONCLUSION: For both diagnostic and evaluation purposes of the edentulous mandible, the panoramic radiograph is not a reliable radiographic technique unless meticulous precautions are taken for reproducible positioning of the patient in the apparatus. PMID- 9347509 TI - Bone regeneration with a calcium sulfate barrier. AB - OBJECTIVES: Bone defects are a challenge for the dental clinician. As widely accepted in guided tissue regeneration, physically halting soft connective tissue proliferation into bone allows for bone regeneration. This concept is the "osteopromotion principle." The aim of this study was to assess the osteopromoting effect of calcium sulfate as a barrier. STUDY DESIGN: Forty male Sprague-Dawley rats were used. Mucoperiosteal flaps were raised bilaterally at buccal and lingual aspects of the mandible to expose the angles. Next, 5 mm through-and-through bony defects were created bilaterally. On the test side, sterile medical grade prehardened calcium sulfate disks were applied both lingually and buccally to cover the defect. The control side defects were left uncovered. All flaps were sutured closed. Observation times were 3, 9, 18, and 22 weeks. RESULTS: Histologic analysis demonstrated that at 3 weeks all test sites showed partial or complete bone healing. Similar findings were reported for all observation times. The control group showed no bone growth at 3 and 9 weeks and partial bone healing at 18 and 22 weeks. CONCLUSIONS: This study indicates that calcium sulfate barriers can exclude connective tissues, allowing bone regeneration during healing. PMID- 9347511 TI - Implant site assessment using panoramic cross-sectional tomographic imaging. AB - OBJECTIVES: The purpose of this study was to evaluate the ability of two different panoramic imaging systems to produce cross-sectional images with accurate vertical dimensions of the posterior mandible. STUDY DESIGN: Three partially edentulous human cadaver mandibles were used for this study. On each mandible, three potential implant sites were arbitrarily identified in an area between the mental foramen and the ascending ramus. Each site was imaged using two different panoramic machines. Using each image, the mandible's outline, cortical thickness, and position of the mandibular canal were traced on clear acetate film. The mandibles were then sectioned at each site to serve as a gold standard. The cadaver sections and tracings (corrected for magnification) were measured, recording the overall mandibular height, distance from the crest of the ridge to the superior aspect of the mandibular canal, and the thickness of the cortical bone at the most inferior aspect of the mandible. RESULTS: There were no significant differences between either of the system's image measures and the gold standard when considering the distance between the crest and the mandibular canal. Differences were noted between the systems measures and the gold standard in the assessment of the cortical bone thickness and the overall mandibular height. CONCLUSIONS: Both imaging systems can be useful for vertical measurements of a potential implant site in the posterior mandible. PMID- 9347512 TI - Influence of the addition of restorations on the diagnosis of caries from digitized bitewing radiographs. AB - OBJECTIVES: The purpose of this study was to assess the influence of additional caries and restorations on the detection of caries on the same radiograph. STUDY DESIGN: Six participants examined five series of four radiographs in which natural carious lesions were present. Each series consisted of the same image with progressively more restorative treatment digitally painted on. The films were randomly presented to the observers who examined the films for the presence and depth of carious lesions. The observers were not informed that the 20 films were disguised versions of the same original five radiographs. RESULTS: The number of carious lesions reported by the six observers did not increase despite the apparent increased restorative intervention viewed on the radiographs. CONCLUSIONS: The complexity of restorative care does not affect observers' ability to correctly detect approximal carious lesions. PMID- 9347513 TI - Diagnostic imaging for a case of maxillary myxoma with a review of the magnetic resonance images of myxoid lesions. AB - The findings of conventional radiography, computed tomography, and magnetic resonance imaging are reported for an odontogenic myxoma arising in the left anterior maxilla of a 50-year-old man. The magnetic resonance imaging characteristics of an intraosseous myxoma are described for the first time. The initial conventional radiographic examination disclosed a unilocular radiolucency with poorly delineated margins as typically seen in malignant tumors. Subsequently, acquired computed tomography scans displayed bony expansion and thinning of cortices on the labial aspect of the lesion. Magnetic resonance imaging revealed a well-defined, well-enhanced mass lesion with homogeneous signal intensity on every pulse sequence. The lesion showed intermediate signal intensity on the T1- and T2-weighted images. Magnetic resonance imaging of the present maxillary myxoma revealed a higher signal intensity on T1-weighted and a lower signal intensity on T2-weighted images than for previously reported myxomas of the soft tissues. This discrepancy might be related to the viscosity of the mucoid substance or the protein density of the tumor. PMID- 9347514 TI - Antibody responses to Rhoptry-Associated Protein-1 (RAP-1) of Plasmodium falciparum parasites in humans from areas of different malaria endemicity. AB - Plasma IgM and IgG antibody reactivities against the recombinant Plasmodium falciparum protein, Rhoptry Associated Protein-1 (rRAP-1) were measured by ELISA in individuals from Sudan, Indonesia, Kenya and The Gambia living in areas of different malaria endemicity. IgG and IgM reactivities to rRAP-1 increased with malaria endemicity. IgG reactivities were associated with spleen rates in Indonesia with high malaria endemicity while IgM reactivities were associated with spleen rates in Kenya with low malaria endemicity. IgG and IgM reactivities to rRAP-1 increased during acute episodes of P. falciparum malaria in Sudanese adults and IgG reactivities remained high one month after treatment in all adults tested. Antibody reactivities to rRAP-1 in Gambian children in the dry season were higher in children with parasitaemia than in children without detectable parasitaemia. Antibody reactivities were not associated with protection against clinical episodes in the following rainy season but higher antibody reactivities were detectable at the end of the rainy season. There was no difference in antibody reactivity to rRAP-1 between Gambian children with mild or severe malaria. PMID- 9347515 TI - An association between activity of the Na/K-pump and resistance of Schistosoma mansoni towards complement-mediated killing. AB - The Na/K-pump or Na+/K(+)-ATPase (EC 3.6.1.37), couples the hydrolysis of ATP to the active transport of Na+ and K+ ions across the plasma membrane of virtually all animal cells. The relationship between activity of the Na+/K(+)-ATPase and the sensitivity of Schistosoma mansoni to immunological attack has been investigated. It has been observed that ouabain, the specific inhibitor of the pump, via a synergistic effect with specific antibody and complement, affects the average membrane potential causing depolarization and death of complement resistant parasites. Thus, apparently, there is association between the inhibition of the Na/K-pump and the lysis and death of the complement-resistant parasite. PMID- 9347516 TI - Host immune response evasion strategies in Ornithodoros erraticus and O. moubata and their relationship to the development of an antiargasid vaccine. AB - We analysed in mice why the salivary gland extract (SGE-2) from Ornithodoros erraticus and O. moubata induce a protective response with Freund's adjuvants (FAs) in swine while the saliva, in natural conditions, does not. Such protection has been ascribed to the fact that administration of SGE-2 plus FAs permits the recognition of certain salivary components that under natural conditions are not immunogenic. The present findings confirm this hypothesis since in mice, which are unable to recognize the above components, the SGE-2-FAs do not induce any protection. We rule out the possibility that the cause of this could lie in the absence of prostaglandin E2 in the SGE-2 (vs saliva) since it is not present in either fluid. Neither could it be due to a change in antibody isotype since those induced by parasites bites and by the SGE-2-FAs are the same (IgG2a > IgG1 > IgG2b; not IgG3, IgM, IgE). No IgG2a were seen when the SGE-2 were administered alone or with alum or ricin. It is therefore suggested that first responses would be Th1 and the second ones Th2, although no IgE is seen in the latter responses either. The parasites do not require complement to feed; by contrast, they block its activation and skin cellular infiltrates, such as those elicited by IL-8, MCP 1 and C5a, do not affect them, regardless of the presence or not of antitick antibodies. PMID- 9347517 TI - Anaplasma marginale: effect of the treatment of cattle with an interferon gamma neutralizing monoclonal antibody or the nitric oxide synthetase inhibitor aminoguanidine on the course of infection. AB - Cattle undergoing initial infection with the rickettsia Anaplasma marginale were treated with either a monoclonal antibody (MoAb) with neutralizing activity for bovine interferon gamma (IFN-gamma) or aminoguanidine (AG), a specific inhibitor of the inducible form of nitric oxide synthetase (iNOS). Plasma levels of MoAb and AG were measured over the time of administration. The course of A. marginale infection was not altered in the MoAb-treated cattle relative to untreated controls. In cattle treated with AG however, A. marginale infection was significantly ameliorated, as judged by lower parasite levels and decreased anaemia in these cattle relative to the controls. The implications of these findings in relation to the basis for immunity against this economically important haemoparasite are discussed. PMID- 9347519 TI - Antigenic variation during malaria infection--the contribution from the murine parasite Plasmodium chabaudi. AB - P. chabaudi AS strain in laboratory mice provides an accessible and useful model for investigating antigenic variation in malaria parasites. Evidence that P. chabaudi AS undergoes antigenic variation is summarized. A live indirect fluorescent test (IFAT) detects a variable antigen on the surface of schizont infected erythrocytes. Five different variable antigen types (VATS) (detected using the live IFAT) are described from a cloned mosquito transmitted parent population. Even during the rising primary parasitaemia VATS switch at high and variable rates (1.2-1.6%). Work towards cloning genes coding for the variable antigen is briefly summarized. Acquired immunity to blood-stage P. chabaudi AS is initially mediated through Th1 CD4+ T cells and after the primary parasitaemia there is a switch to Th2 CD4+ T cells. Acquired immune effector mechanisms are discussed in the context of antigenic variation by the parasite. PMID- 9347518 TI - Polyclonal T-cell responses to Plasmodium falciparum gametocytes in malaria nonexposed donors. AB - Human peripheral blood T-cells mount vigorous proliferative responses to asexual stage parasites of P. falciparum regardless of whether the donor has been exposed to the parasite. Here using highly purified P. falciparum gametocytes we show that the same is also true for this stage of the parasite. Gametocytes, like immature trophozoites, preferentially activate CD4+ T cells gamma delta T-cell activation, commonly observed in response to mature schizonts or supernatants from asexual-stage cultures, does not occur. Furthermore, the CD4+ T-cell receptor variable region (TCRV beta) usage to those stimulated by asexual parasites and there is no preferential usage of TCRV beta elements. The CD4+ T cell precursor frequencies for gametocyte and asexual trophozoite responses are remarkably similar. 'Cross-reactivity' of gametocytes and asexual stage responses was confirmed by selective depletion of cells responding to particular stages of the parasite using the cytostatic drug cytosine arabinoside (Ara-C). These results suggest that the CD4+ T-cell responses from malaria non-exposed donors are common to gametocytes and asexual trophozoites. PMID- 9347521 TI - Spoken messages as auditory cues for orientation in promoting indoor travel and activity by persons with multiple disabilities. AB - Spoken messages lasting about 2 sec. and occurring every 8 or 9 sec. were used as auditory cues for orientation to guide two persons with multiple disabilities to destinations for indoor activities. The use of spoken messages was alternated with buzzer-like sounds of the same duration and frequency of occurrence. The preliminary data indicated that both the spoken messages and the buzzer-like sounds were very effective as orientation cues. Staff found no difference in disturbance between them, yet, both were considered less disturbing than buzzer like sounds of previous studies which had a higher frequency of occurrence. PMID- 9347520 TI - Emergent properties following brain injury: the claustrum as a major component of a pathway that influences nociceptive thresholds to foot shock in rats. AB - Flinch (pain) thresholds for electric current delivered to the feet were correlated with the amount of necrosis within the diencephalon and telencephalon for rats in which seizures had been induced by lithium and pilocarpine about two months before the testing. The shared variance of the quantitative damage within the claustrum, the anterior part of the paraventricular nucleus of thalamus, (central) mediodorsal thalamus, and lateral amygdala (ventromedial part) explained 81% of the variance in the nociceptive (flinch) thresholds. A primary role of the claustrum within the neuropathways that mediate the response to the interoceptive and "painful" characteristics of stimuli is indicated. The concept of primary pathways versus "emergent" pathways subsequent to excitotoxic damage within the neuromatrix is discussed. PMID- 9347522 TI - Effect of information about odor on causal ascriptions for illness. AB - The effect of information about detection of an odor on causal ascriptions for illness was investigated. In four different scenarios perceptions regarding the cause of a hypothetical symptomatic experience were compared for events described with and without an odor. Participants (N = 106) were asked to imagine themselves becoming ill after engaging in several common experiences, including pumping gasoline at a service station. In two scenarios participants read that they smelled an offensive odor while pumping the gasoline whereas in two other scenarios no information about an odor was provided. Further, information about gasoline described with or without odor was presented either early or late in the stimulus paragraphs. All participants then responded to questions including an open-ended question asking them to make causal attributions for their illness. Participants in the odor-suggested group ascribed the cause of illness more frequently to gasoline and perceived the probability of other potential causes as lower than did participants in the nonodor suggested group. Findings suggest that peoples' implicit theories about toxicity contain causal connections between malodorous stimuli and illness. The implications of implicit theories for perception of illness are discussed. PMID- 9347523 TI - Perceptions of motivational climate, perceived competence, and motivation of students of varying age and sport experience. AB - This study examined differences in students' motivation in Greek physical education classes depending on age and amount of experience in sport and the extent to which these differences reflected divergent perceptions of competence and classes' motivational climate. 1,393 students responded to questionnaires measuring motivational climate, perceived competence, preference for challenge, interest in the lesson, and perceived importance of the lesson. Students who were not involved in out-of-school sport activities had lower scores on perceived physical competence, perceived learning orientation of the class, preference for challenge, interest in the lesson, and perceived importance of the lesson than students who were involved in organized sport. These differences in students' motivations decreased when scores on perceived learning goals and perceived physical competence were adjusted. Senior high school students (16 yr. old) were much less motivated than junior students (13 yr. old), but these differences were decreased or eliminated when scores on perceived learning orientation were controlled. These results suggest that to increase all students' motivation in physical education, a strong emphasis on personal progress should be adopted. PMID- 9347524 TI - Reliance on visual imagery and its relation to mental rotation. AB - The relationship of habitual use of visual imagery and mental rotation was investigated. Reliance on Visual Imagery scores were used to define subjects as high frequency or low frequency visualizers. During the mental rotation task, subjects indicated if a pair of 2-dimensional stimulus figures displayed on a computer screen were identical or mirror-images. Figures on the right were rotated in relation to those on the left by 0, 60, 120, or 180 degrees. Data supported the prediction that subjects who report high use of imagery would perform the task with greater accuracy (z = 1.97, p < .05) than subjects who reported low use. The imagery groups did not differ in response latency (z = .91, p < .36). A comparison of performance on Trials 1 to 24 with performance on Trials 115-138 indicated a learning effect in both accuracy (z = 7.58, p < .01) and latency (z = 9.72, p < .01) for all subjects. PMID- 9347525 TI - Fractionated reaction time as a function of magnitude of force in simple and choice conditions. AB - The present study examined whether varying magnitude of force required to perform an isometric response influences fractionated reaction time in simple and choice conditions and whether reaction time and premotor time to initiate the response are shorter when force is selected freely by the subject than when it is selected by the experimenter. 20 subjects were required to react and produce a designated peak force as quickly and accurately as possible by squeezing a handle after a reaction signal. Four different magnitudes of force were 30, 50, and 70% of the maximum grip strength of the subjects and subject-selected magnitude of force. Reaction time and premotor time did not change across the range of forces examined in both simple and choice reaction-time conditions regardless of whether a desired force was selected by the experimenter or by the subject. These findings suggest that programming an isometric response may require a constant amount of time. PMID- 9347526 TI - Reported prevalence of unconsciousness from mechanical impact to the head in university populations during a fifteen-year period. AB - The prevalence of at least one episode of unconsciousness during childhood due to a mechanical impact to the skull was inferred by the response to one item embedded within a questionnaire of 140 items. 50% of the 633 university men and 33% of the 863 university women reported such unconsciousness; the prevalence did not change significantly between samples over a 15-yr. period. Multiple regression analysis indicated that the 10 items most strongly associated with the report of childhood unconsciousness did not explain more than about 10% of the variance. The majority of the items were those associated with complex partial epileptic-like signs and included (adult) episodes of memory blanks, mystical experiences, dissociation, and sudden meaningfulness. PMID- 9347527 TI - Ear dominance and telephone sales. AB - In a field study, three equally sized sales teams used on of three head-sets- left, right, both ears--for a day's selling of insurance by telephone. This had no effect on sales. In a retrospective study of records, daily sales performance including the percentage conversion rate for sales divided by the number of calls and the number and duration of calls was related to preference for type of head set. Sales were markedly influenced by the choice of head-set. People who chose to wear the left earpiece significantly out sold the others wearing right and stereohead-sets. Neither the number of incoming calls nor the time spent on the telephone were influenced by the choice of head-set. When sales are analysed in terms of individual differences in personal preference for type of head-set, those who chose the left ear had an advantage. Forced use of the left, versus right ear or both ears for one day had no effect. PMID- 9347528 TI - Economic development, suicide and homicide. PMID- 9347529 TI - Speakers' sex differences in voice onset time: some preliminary findings. AB - This study presents a brief investigation into sex differences of speakers in the voice onset time of English plosives that are stressed in both word-initial and prevocalic position. 72 short phrases were presented to 5 men (range 25 to 37 years, mean age 34.2 yr.) and five women speakers (range 28 to 38 years, mean 32.6 yr.). Analysis showed that the women as speakers had on average, longer voice onset time values than their male peers. PMID- 9347530 TI - Performance of soccer players on tests of field dependence/independence and soccer-specific decision-making tests. AB - The purpose of this study was to examine the relationship of the performance of male amateur soccer players on tests of field dependence/independence and soccer specific decision-making tests. The relationships between the participants' (N = 14) accuracy, and speed of decision, on simple and complex soccer decision-making tests; scores on Parts B or C of the Group Embedded Figures Test under normal conditions: scores on Parts B or C of the Group Embedded Figures Test when timed; and time taken to complete the timed condition of the Group Embedded Figures Test were examined. There were no significant correlations between performance on the soccer specific tests and the tests of field dependence/independence. PMID- 9347531 TI - Reading, writing, and vocabulary skills of children with strokes due to sickle cell disease. AB - The reading, writing, and vocabulary skills of 8 children with strokes due to sickle cell disease were compared with 8 control children. The former were delayed in reading and writing skills but not in vocabulary development or use. PMID- 9347532 TI - Effects of circadian orientation, time of day, and arousal on consumers' depth of information processing of advertising. AB - Since depth of information processing, as defined by MacInnis and Jaworski in 1989 has been shown to influence the strength of the relation between the intent to purchase and the attitudes toward the advertisement, this paper focused on the interactive effects of three antecedents of information processing, arousal, circadian orientation, and time of day (Morning vs Evening). Analysis indicated that deeper information processing is reached by 65 morning-oriented consumers who are exposed to advertisements in the morning and by 52 relaxed consumers who are exposed to advertisements in the evening. Theoretical explanations and managerial implications are proposed. PMID- 9347533 TI - Assessment of hearing-impaired athletes' anxiety and self-perceptions. AB - 70 hearing-impaired basketball players participating in a national basketball tournament completed Neeman and Harter's Self-perception Profile examining their feelings of social acceptance, athletic competence, and global self-worth. In addition, players completed the Sport Competition Anxiety Test for trait anxiety and the Competitive State Anxiety Inventory evaluating their cognitive and somatic anxiety as well as their feelings of self-confidence. Correlations indicated an inverse relationship for subjects' ratings of athletic competence with their scores on trait anxiety and rated cognitive and somatic state anxiety. The correlation between rated self-worth and the subjects' feelings of confidence was low and positive. Results are discussed in relation to achievement-motivation theory. PMID- 9347534 TI - Determinants of choice of method for suicide and the person/situation debate in psychology. AB - Researchers have not identified personological predictors of choice of method for suicide, whereas an availability hypothesis has received support. Thus, the situational position may be useful as a framework for efforts to prevent suicide. PMID- 9347535 TI - Mental chronometry with simple linear regression. AB - Typically, mental chronometry is performed by means of introducing an independent variable postulated to affect selectively some stage of a presumed multistage process. However, the effect could be a global one that spreads proportionally over all stages of the process. Currently, there is no method to test this possibility although simple linear regression might serve the purpose. In the present study, the regression approach was tested with tasks (memory scanning and mental rotation) that involved a selective effect and with a task (word superiority effect) that involved a global effect, by the dominant theories. The results indicate (1) the manipulation of the size of a memory set or of angular disparity affects the intercept of the regression function that relates the times for memory scanning with different set sizes or for mental rotation with different angular disparities and (2) the manipulation of context affects the slope of the regression function that relates the times for detecting a target character under word and nonword conditions. These ratify the regression approach as a useful method for doing mental chronometry. PMID- 9347536 TI - Finno-Ugrians and suicide. PMID- 9347538 TI - Lateralization in appreciation of humor: sex differences vs stimulus effects. AB - The finding that women rate funnier humorous items with left-ear input, while men give higher ratings with right-ear input has been cited as evidence for a biological basis for sex differences in appreciation of humor. However, in 1991 Gallivan did not find this effect and suggested that the earlier finding could have been due to the use of 'male-oriented' stimuli. In this study, 72 subjects rated the funniness of 32 'female oriented' comedy excerpts, presented monaurally. Women gave higher ratings with right-ear input but men's ratings were not affected by ear of presentation. These findings represent another failure to replicate the earlier-reported hemispheric effect and support the conclusion that it may have been due to the stimuli used. PMID- 9347537 TI - Age differences in remote pointing performance. AB - The purpose of this study was to investigate age differences in remote pointing movements. The subjects were recruited from three age groups (ages 18-22 yr., 40 50 yr., and 60-70 yr., with 9 men and 9 women in each group). They were required to perform cursor-positioning tasks using a remote pointing device, in which the dependent measures were the time taken to reposition the cursor and the accuracy of subjects' movement trajectories. The movement time was further separated into two components, First Submovement duration and Adjustment Submovement duration. Analysis indicated that age groups showed reduced performance on remote pointing. Moreover, remote positioning movement for the young-adult group was mostly completed in their First Submovement phase, while the elderly subjects spent most of their movement time on the Adjustment Submovement phase. These results support the proposition that different age groups exhibit different kinds of movement patterns. PMID- 9347539 TI - Review of the criterion-related validity of the WISC-III: the first five years. AB - A review of studies comparing the WISC-III to other measures of intelligence and achievement indicates the revision is continuing to meet the standards established by its predecessor, the WISC-R. Correlations are substantial, both for measures of ability and achievement; however, compared to the WISC-R, the WISC-III Full Scale IQ tends to be about six points lower, probably reflecting the "Flynn effect" (1984). As a consequence, eligibility for programs in special education may be negatively affected. PMID- 9347540 TI - Perceived traits of male and female athletes. AB - Profiles of personality traits for male and female athletes were obtained from 133 men and 71 women raters. Traits were rated using a 7-point semantic differential with 11 bipolar items. A profile analysis showed that the profiles of the traits were distinct. There were no significant differences in the ratings by men and women raters. Male athletes were rated as more active, aggressive, competitive, dominating, controlling, instrumental, and public. Female athletes were rated as more goal-oriented, organized, and rule-governed. PMID- 9347541 TI - Effects of figure context on the apparent length of a line. AB - This study investigated the effects of figure context on the apparent length of a line. In Exp. 1, ten participants were asked to adjust the length of a comparison line to match a standard line enclosed within a rectangle. The participants consistently overestimated the length of the standard line, demonstrating the stretching effect of figure context on the apparent length of a line. In Exp. 2 (12 participants), the size of the context figure was varied and it had no significant influence on the magnitude of the context effect. In Exp. 3 (nine participants), the context effect was shown not only for squares and rectangles but also for other shapes of figures such as circles and 5-pointed stars. We discuss the possible mechanism of the figure-context effect within Gregory's (1970, 1978) misapplied constancy theory of visual illusions. PMID- 9347542 TI - Language anxiety and proficiency in a foreign language. AB - Study examined the extent to which there would be differences in oral and written proficiency in a foreign language among groups of low-, average-, and high anxious high school students. Participants were 60 girls attending a single-sex, college-preparatory high school and completing the second year of a foreign language course. Analysis showed over-all differences on measures of proficiency in the foreign language among the three groups. The results support the hypothesis that anxiety about foreign language learning is likely to represent students' differences in language learning. PMID- 9347543 TI - Evidence for psychological refractory effect in motor inhibition for a dual response Go/No-Go task. AB - Human subjects exhibit difficulty in initiating two independent, discrete responses in close succession, a difficulty known as the 'psychological refractory effect.' It is not yet known whether motor-inhibition processes are under the influence of this effect, as are motor-execution processes. This study examined the temporal changes of subjects' reaction times, interpreted in terms of motor programming for inhibition, in a dual-response Go/No-Go task that required two independent responses in close succession. Eight subjects performed the task with both a shorter (400 msec.) and a longer interstimulus interval (800 msec.). The mean reaction time for the second stimulus (RT2) in the Go response of the 400-msec. condition was significantly longer than that of the 800-msec. condition. For committed error responses during the No-Go trials, the mean RT2 in the 400-msec. condition was longer than that in the 800-msec. condition. The total number of these errors in the 400-msec. condition was significantly greater than that in the 800-msec. condition. These results suggested that both the motor execution processes and motor-inhibition processes were influenced by the psychological refractory effect. PMID- 9347544 TI - Lateral preferences in a German population. AB - There are only a few studies in the literature concerning all four lateral preferences (handedness, footedness, eyedness, earedness) and presenting their interrelationships. Porac and Coren's 1981 inventory was used to assess hand, foot, eye, and ear preference in a German sample of 506 men and 430 women. A right sided preference was found for hand, foot, eye, and ear preference among 91%, 74%, 66%, and 63% of the sample, respectively. More men were left-handed and left-footed than women. There were no significant sex differences for eyedness and earedness. Correlations ranged from .22 between hand and eye laterality to .44 between hand and foot laterality. PMID- 9347545 TI - The organization of visual objects: randomness. AB - A method was described for determining whether or not a pattern of visual objects is arranged randomly. Based on distance to nearest neighbor, it includes three conditions and is designed to avoid Type I errors. Examples were given, and comparisons were made with other methods. PMID- 9347546 TI - Isolated sleep paralysis, vivid dreams and geomagnetic influences: II. AB - This report describes a test of the hypothesis that significant changes in the ambient geomagnetic field are associated with altered normal nighttime dream patterns. Specifically, it was predicted that there would be a greater incidence of isolated sleep, paralysis or vivid dreams with abrupt rises and falls of geomagnetic activity. The author's (JC) and a second subject's (KC) daily reports of dream-recall were analyzed in the context of daily fluctuations of geomagnetic activity (K indices). Two analyses of variance indicated (i) significantly higher geomagnetic activity three days before a recorded isolated sleep paralysis event and (ii) significantly lower geomagnetic activity three days before an unusually vivid dream took place. Conversely, geomagnetic activity did not fluctuate significantly for randomly selected days. Testing a large sample over time is required for confirmation and extension of this work. PMID- 9347547 TI - Ethnicity, sex and snoring. AB - The associations of the incidence of snoring with ethnicity and sex were measured using self-report data collected from 1098 university undergraduates. Both the relationships were significant. The unique feature of these data is that for the first time, a significant relationship between ethnicity and snoring has been reported. PMID- 9347548 TI - Veering in women: inconsistency of forward and backward progression. AB - In a large rectangular room, 13 blindfolded women attempted to walk in a straight line from one end of the room to a target centered at the other end of the room. On 12 trials, the women walked forward, and on 12 trials they walked backward. On half the trials under each of these conditions, they walked toward the north, and on the other half to the south. Performance errors were highly correlated for northward and southward progression, an indication of good reliability for this veering task. Veering during forward progression was not significantly related to veering during backward progression. Individual consistency in veering was demonstrated in several ways, and approximately half the participants veered in the same direction on nearly all trials. These results indicate that veering should be considered as an additional manifestation of lateral preferences in human motor behavior. PMID- 9347549 TI - Public school nurses' knowledge of sickle cell disease. PMID- 9347550 TI - Relative importance of success in sport and schoolwork. AB - Cross-domain studies of achievement-related cognitions have been gaining attention in the United States. This study provides comparative data from a United Kingdom upper school. 390 subjects age 13 (n = 218) or 15 (n = 172) years rated the importance of success in sport and schoolwork. Academic success was more important than sport success, and sport success was more important to boys (n = 203) than girls (n = 187). Academic success was less important to older subjects, and sport success was less important to older girls. Predictions, from importance and perceived ability, of free time spent in each domain were stronger for sport than for schoolwork indicating that the model held better for voluntary activity. PMID- 9347551 TI - Recall of the visual body image using a novel boundary detection task. AB - A new task for eliciting a pictorial mental image of the body or other objects is described. The task involved relating a pair of crosses to the boundary of a mental image 'projected' onto a computer screen. Responses were assessed for accuracy defined as identifying a relationship between a cross and an image that would hold when a photograph (of the same object) was substituted for the mental image. A group of 30 female students achieved between 70 to 80% accuracy when using this task to assess mental images of their own faces, torsos, or a familiar nonbody object. Accuracy was similar for body and nonbody objects. The presence of some kind of quasipictorial representation of the body is confirmed. Its characteristics await further elucidation. PMID- 9347552 TI - Implicit and explicit tests: evidence for dissociable motor skills in probable Alzheimer's dementia. AB - Previous authors reported evidence for intact implicit memories (those retrieved without conscious effort) in a serial reaction time task for both Alzheimer's subjects and age-matched controls, although performance on an explicit memory task (requiring conscious effort for retrieval) was poor. The current study assessed latencies on a puzzle-assembly task to assess implicit (procedural) memory for 23 female volunteers. Nine participants suffered from probable Alzheimer's Disease and fourteen did not. Even when subjects had no explicit memory of practicing the task, they demonstrated savings upon relearning. Implications for research on memory dissociations in Alzheimer's Disease are discussed. PMID- 9347553 TI - Childhood Tourette's syndrome and the Thematic Apperception Test: is there a recognizable pattern? AB - 26 children, one-half of whom were diagnosed with Tourette's syndrome, were administered the Thematic Apperception Test. A pilot investigation suggested that children with Tourette's syndrome produced high rates of responses in four categories: references to physical aggression, supernatural power, character names, and specific quantities. When compared with other children treated for emotional and behavioral problems at the same facility and matched on demographic variables and IQ, children with Tourette's syndrome were not more likely to produce Thematic Apperception Test responses in any of the four categories. This study shows that a recognizable pattern on projective testing was not easily established among these 13 children with Tourette's syndrome and further highlights the importance of using mental health comparison groups when investigating clinical disorders. PMID- 9347554 TI - Welfare and suicide in 18 affluent capitalist democracies. PMID- 9347555 TI - Description of phonological patterns for nineteen elementary-age children with hearing losses. AB - The speech productions of 19 hard of hearing children between 5 and 12 years of age were examined for errors related to phonological process categories. For comparison, the subjects were divided into groups of 9 with Profound and 10 with Moderate to Severe hearing losses. There was a significant relationship between hearing loss and phonological errors. Seven phonological processes were evident in at least 33% of obligatory contexts. Prevalent processes included final consonant deletion and cluster reduction. The most prevalent deficiencies included /r/ and /l/ phonemes. Subjects with Profound hearing losses produced more errors over-all as well as more errors in each phonological process category. Subjects with Profound hearing losses frequently deleted entire consonant clusters, whereas subjects with Moderate to Severe hearing losses did not. Results are also discussed in relation to normal development. PMID- 9347556 TI - Social characteristics of states which do not prohibit assisted-suicide. PMID- 9347557 TI - Age, side height, and spindle shape of the crib in climbing over the side. AB - The crib is the only infant product in which a consumer, such as a parent or caretaker, is encouraged to leave the infant unattended, usually alone in the bedroom, while the infant is sleeping or going to sleep or waking. Given frequent falls from the crib, federal crib regulations have set the minimum distance between the top of the mattress support and top of the crib side rail as 26 in. and this height must include a 6-in. thick mattress. When a mattress is used with the crib, the actual height of the crib side as a barrier is 20 in. These crib regulations also require instructions for the caretaker to discontinue using the crib when the child's height is 35 inches. These federal crib regulations attempt to create an escape-resistant sleeping environment for all children who are less than 35 in. tall. Of 144 children between the ages of 12.5 and 36.5 mo. observed while attempting to climb out of a crib, who also had standing heights of less than 35 in., many were able to climb from cribs with side rail heights of 26 in. When the crib side-rail height was raised beyond the minimum of 26 in., the frequency of children climbing over the crib rails decreased. PMID- 9347558 TI - Relations of scores on Children's Embedded Figures Test with age, item difficulty and internal consistency. AB - This study analyzes the Children's Embedded Figures Test by examining its internal consistency, test-retest reliability, the order of difficulty of the items, and the change of scores with age. Among the sample 337 boys and 287 girls who were between the ages of 6 and 11 years and in the first five grades of primary school scores increased significantly. The test presented moderate internal consistency (.86), and the test-retest reliability after one year was .63. The order of difficulty of the items did not coincide with the order proposed by the test's authors and varied from grade to grade, i.e., in the Tent series Item 4 and in the House series Item 5 were among the most difficult. PMID- 9347559 TI - Experimental induction of the "sensed presence" in normal subjects and an exceptional subject. AB - 9 of the 15 volunteers who were exposed to successive 3-min. durations of bursts of different types of weak (1 microT) complex magnetic fields or sham-fields reported the sense of a presence as indicated by a button press at the time of the experience. Reports of subjective experiences indicated that attempts to "focus" cognitively upon the location of the presence altered its location or induced its "movement." An exceptional subject who had a history of experiencing within his upper left peripheral visual field "flashing images" concerning the health and history of people [when handling their photographs] was also exposed to the burst sequences. Numbers of button presses associated with the experiences of a mystical presence, to whom the subject attributed his capacity, increased when the complex magnetic fields were applied without the subject's knowledge. The results support the hypothesis that the sense of a presence, which may be the common phenomenological base from which experiences of gods, spirits, angels, and other entities are derived, is a right hemispheric homologue of the left hemispheric sense of self. PMID- 9347560 TI - Scores on emotional self-control of elementary school children after a five-week running program. AB - This study tested the prediction that fitness running would increase elementary school children's emotional self-control (Grade 3, n = 26, Control, n = 25). The results were contrary to the hypothesis. PMID- 9347561 TI - An exploration of the influence of educational placement on the community recreation and leisure patterns of children with developmental disabilities. AB - This study explored the recreation and leisure patterns of children with moderate to severe developmental disabilities, 28 parents of children labeled "trainable mentally handicapped" were questioned about the type of sport, exercise, and social activities their children participated in as well as about the opportunities for social inclusion with nondisabled peers that these activities afforded their children. Analyses indicated that most of the children participated in segregated recreation and leisure activities. However, after adjusting for age, an analysis of covariance showed that children placed in more socially integrated educational settings participated in significantly more inclusive recreational activities than children who were served in segregated educational settings. PMID- 9347562 TI - Speed and accuracy of eye-gaze pointing. AB - The current experiment examined the speed-accuracy trade-off of saccadic movement between two targets. Ten subjects looked alternately at two targets as fast and as accurately as possible for 2 min. under different conditions of target size, distance between targets, and direction of eye movement. Saccadic movement of the left eye was tracked and recorded with an infrared eye monitoring device to compute the starting position, ending position, and duration of each saccadic movement. Eye-movement time was significantly related to target size and distance between targets, but the speed-accuracy trade-off was significantly different from that predicted by Fitts' Law. Reaction time was not significantly changed by the direction of eye movement. PMID- 9347563 TI - Differential threshold of length and response criterion for inspecting irregular objects. AB - This research investigated human visual sensitivity and bias in inspecting irregular objects. A preliminary study was conducted using the method of constants to determine the threshold value for judgment of size. A factorial experiment was conducted using payoffs, rate of defective items, and detectability in the signal-detection theory as the factors. In total, eight experimental conditions were tested. 10 college students were recruited as subjects. Each subject was asked to compare 40 teapot shapes to a standard teapot shape under eight experimental conditions. Defective shapes were generated by lengthening the vertical dimension of a standard teapot shape by a factor of 1.01 and 1.04 for 'low' and 'high' detectability. The decision time and responses of 'identical' or 'different' were collected under all experimental conditions. Analysis indicates that the decision-making strategy used to inspect this irregular object was very close to maximizing the accuracy of decision-making by considering the rate of defective items. This result is different from most research findings in signal-detection theory in which responses of human beings are similar to degraded Bayes optimizers. The standard deviation of the signal distribution was about 1.30 and 1.41 times that of the noise distributions for 'low' and 'high' detectability. PMID- 9347565 TI - Verbal-to-manual and manual-to-verbal dual-task interference in left-handed and right-handed adults. AB - Verbal-manual interference was investigated with 80 students who were divided into four groups by sex and hand preference. Unilateral finger-tapping was measured during no-load conditions and during two concurrent tasks of word reading (aloud) and sentence reading (silent). During concurrent tasks, no selective interference effects for the preferred hand were found; however, when participants were classified according to consistent handedness instead of hand preference, consistent right-handers exhibited selective right-hand tapping interference during concurrent word reading, whereas consistent left-handers showed generalized interference. During concurrent sentence reading, men showed selective right-hand interference, irrespective of handedness. The influence of tapping on word reading was also examined. Concurrent tapping lowered word reading performance substantially, showing that finger-tapping and word reading interfered reciprocally. PMID- 9347564 TI - Internal consistency of manual muscle testing scores. AB - The internal consistencies of manual muscle test scores of the actions of three upper and three lower extremity muscles were examined among 37 home care patients. The correlations between scores of specific pairs of actions ranged from .01 to .88. Cronbach alphas ranged from .59 to .88. Manual scores of limb muscle strength, therefore, appear to possess suitable internal consistency. PMID- 9347566 TI - Personality structure of elderly drivers. AB - This paper reports the factor structure of a 37-item personality questionnaire intended to be predictive of driving performance in elderly persons. Subjects were 191 persons 63 years of age of older, about half of whom also were given perceptual/cognitive tasks and drove on a closed driving course. Although the personality questionnaire did not predict driving skill, the factor structure of the questionnaire is of interest. Of several factor analyses, the most satisfactory was a 2-factor solution. We interpreted the approximately orthogonal factors as measuring what we labeled Competence and Emotionality. PMID- 9347567 TI - Perceptions of high risk sports. AB - High risk sports were rated as to risk, appeal, and likelihood of participation by 282 men and 162 women. Ascending order of perceived risk was skiing, scuba diving, bungee jumping, rock climbing, motorcycle racing, hang gliding, cliff jumping, and skydiving. Profile analysis showed stated likelihood of participation to be directly related to appeal and inversely related to perceived risk. PMID- 9347568 TI - Changes in perceived color with intermittent illumination. AB - Using 24 observers with normal color vision, perceived shifts in hue were determined for a yellow-red, green, and blue-green at intermittencies of 5, 10, and 20 cps. The hue shift for yellow-red was consistent with the hue shift exhibited by a deuteranomalous observer while the hue shift for green and blue green was consistent with that exhibited by a protanomalous observer. PMID- 9347569 TI - Comparison of preschool children's scores on the Modified Version of the Bender Gestalt Test and the Developmental Test of Visual-Motor Integration. PMID- 9347570 TI - Molecular modeling study of the multidrug resistance modifiers cis- and trans flupentixol. AB - Recent drug-membrane interaction and quantitative structure-activity relationship studies of thioxanthenes and related compounds acting as multidrug resistance (MDR) modifiers pointed to the importance of the stereoisomery for their MDR reversing activity. Therefore a molecular modeling study of trans-(T) and cis flupentixol (C) was performed in order to elucidate the observed discrepancy between equal binding potency to P-glycoprotein and different MDR reversing activity of the two stereoisomers. The results show that the 2 to 3-fold difference in MDR reversing activity of T compared to C might be related to a different orientation of the molecules in the membrane lipid environment. From the conformations generated by the SYBYL systematic search procedure those comprising local energy minima were selected and further optimized with semiempirical quantum chemistry methods. From the optimized conformations those that corresponded to 1H NMR results on drug conformations in lipid environment were selected for further molecular modeling studies. The electrostatic and lipophilic fields of T and C were compared in order to identify molecular properties related to the activity difference. The results show that the electrostatic fields of the drugs when similar in shape are dissimilar and that the lipophilic and hydrophilic regions are clearer separated in T in comparison with C. This imposes a better fitting of T compared to C to membrane lipid environment in accordance with the observed higher interaction strength of T with phospholipids. PMID- 9347571 TI - Modified radioimmunoassay for pimozide. AB - Tritium labelled pimozide, the tracer in Michiels' RIA of pimozide, can be substituted for a derivative of pimozide with tritium labelled tag, prepared in a more convenient way than the original tracer. In the synthesis of the immunogen the conjugation rate is optimized to be 12 to 13. PMID- 9347572 TI - Formulation of an aqueous injection of flurbiprofen. AB - Flurbiprofen (1) is an analgesic, antipyretic and antiinflammatory agent which is practically insoluble in water. The aqueous solubility of 1 using various hydrotropes was attempted. The solubility increased up to 63 times in the case of sodium benzoate. Using selected hydrotropes, aqueous injections of 1 were formulated. Formulations were studied for physical and chemical stability. Some of the formulations showed reasonable stability which can be further enhanced by incorporation of appropriate formulation additives. These formulations were evaluated for antiinflammatory and analgesic activity and showed promising results. PMID- 9347573 TI - Nifedipine/atenolol interactions in gastrointestinal absorption and biotransformation. AB - The combination of nifedipine and atenolol is widely used for the treatment of hypertension. In the present study, experiments performed in rats indicated that neither drug affects the gastric or intestinal absorption of the other. In assays of the biotransformation of nifedipine in liver homogenates, breakdown was much more rapid in homogenates from male rats than from female rats. In the presence of atenolol, the breakdown rate was significantly increased in homogenates from male rats, and significantly reduced in homogenates from female rats. PMID- 9347575 TI - Improvement of the biological performance of oral anticoagulant drugs. 2. Dicumarol. PMID- 9347576 TI - Neutrophil pathophysiology. PMID- 9347577 TI - Severe chronic neutropenia: pathophysiology and therapy. AB - The development of recombinant-met human granulocyte-colony stimulating factor (r metHuG-CSF) for clinical use has had a major influence on the treatment of many diseases. This impact has perhaps been greatest for treatment of severe chronic neutropenia (SCN) conditions for which there were no predictably effective treatment before the availability of these growth factors, particularly r-metHuG CSF (Filgrastim, Amgen Inc, Thousand Oaks, CA; or Lenograstim, Rhone-Poulenc Rorer, Milan, Italy). Based on careful studies in many countries it is now known that more than 95% of these patients will respond promptly to r-metHuG-CSF treatment with normalization of the blood neutrophil levels and reduction in the occurrence of both major and minor consequences of their severe neutropenia. The availability of this treatment will undoubtedly lead to much additional research on the mechanisms governing neutrophil production and the basic mechanisms that can cause neutropenia among patients who have SCN. Among patients who have SCN those who are diagnosed to have severe congenital neutropenia (Kostmann's syndrome) or Shwachman-Diamond syndrome are at risk of developing myelodysplasia and/or acute myelogenous leukemia. The role of r-metHuG-CSF in facilitating the risk remains to be determined. Thus, it is important that long-term evaluation of the safety and efficacy of treatment of SCN and cooperation in research on these rare conditions proceed under the auspices of an international registry monitoring the clinical outcome of patients with severe congenital neutropenia. PMID- 9347578 TI - Primary inherited defects in neutrophil function: etiology and treatment. PMID- 9347579 TI - Acquired neutrophil dysfunction and diseases with an inflammatory component. PMID- 9347580 TI - Control of late neutrophil-specific gene expression: insights into regulation of myeloid differentiation. AB - During myeloid differentiation, the pluripotent hematopoietic stem cell passes through several well-defined morphologic stages within the bone marrow. These changes include progressive nuclear segmentation and the acquisition of stage specific granules. Primary granules appear at the myeloblast stage, and are found in both neutrophils and monocytes. At the myelocyte stage, neutrophil precursors acquire specific granules, a marker of commitment to terminal neutrophil differentiation. This complex developmental pathway is just beginning to be elucidated. Current evidence suggests that myeloid differentiation is regulated primarily by transcriptional regulatory proteins, and that dysfunction of those regulators is involved in most disorders of neutrophil maturation. Furthermore, there is evidence that study of late gene expression may provide insights into more proximal events in granulocytic maturation. In this review, we provide a brief overview of myeloid differentiation with emphasis on the culture systems available for the study of granulopoiesis and the insights they provide into the regulation of late neutrophil-specific gene expression. We discuss the relevance of these observations to our understanding of the pathogenesis of defects in neutrophil differentiation. PMID- 9347581 TI - Neutrophil receptors for interleukin-8 and related CXC chemokines. AB - Both the beneficial and harmful roles of neutrophils are critically dependent on the capacity of the cell to undergo directed migration from the blood to local tissue sites. Because the CXC chemokine interleukin-8 (IL-8) is a powerful mediator of this process, its receptor is a reasonable target for development of treatments for neutrophil-mediated inflammation. However, this strategy has been complicated by the discovery of two distinct IL-8 receptors, CXCR1 and CXCR2, that are coexpressed on human neutrophils. Both are 7-transmembrane domain-type proteins functionally coupled to G proteins. Although both receptors bind IL-8 with high affinity, they differ in selectivity for other CXC chemokines, as well as in their regulation and signal transduction, but whether they also differ biologically is not yet clear. PMID- 9347582 TI - Neutrophil adhesion and the therapy of inflammation. AB - Cell adhesion has an essential role in neutrophil recognition of sites of inflammation, migration through endothelium and extracellular matrix, and activation to full effector function at these sites. Adhesion molecules on the neutrophil involved in these events include members of the selectin, integrin, and immunoglobulin superfamilies. Inhibition of neutrophil adhesion holds the promise of interrupting idiopathic inflammation in a variety of diseases, but also can increase susceptibility to infection. The therapeutic challenge is to design agents of sufficient specificity to block the deleterious effects of neutrophil influx and activation without interference with host defense. This will require improved understanding of the molecular events regulating, and regulated by, neutrophil adhesion. PMID- 9347583 TI - Platelet-leukocyte-endothelial cell interaction on the blood vessel wall. AB - Leukocytes, platelets, and endothelial cells interact at sites of vascular injury and inflammation through adhesion receptors on the cell surface. On binding of ligand to receptor, these receptors initiate intracellular signaling that leads to the modulation of several biological properties of the cells involved. These finely regulated processes involve several classes of cell adhesion molecules: integrins, immunoglobulin-like proteins, selectins, and mucin-like proteins as well as an array of soluble mediators. Interaction of these cell adhesion molecules serves to recruit circulating cells to the blood vessel endothelium or to accumulated platelets on the vessel wall and to foster cell-cell communication. The importance of these interactions to inflammation, blood coagulation, and the immune response is outlined. PMID- 9347584 TI - Interactions of neutrophils and coagulation proteins. AB - Some of the interactions between coagulation factors and neutrophils are described. Proteins of the contact system, FXa, thrombin, and fibrinogen all bind to various sites on the neutrophil. This binding has a dual purpose: the assembly of coagulation complexes such as the prothrombinase complex and the contact system on the neutrophil membrane and influencing the various neutrophil functions including chemotaxis, aggregation, degranulation, and transendothelial migration. In addition, neutrophil elastase degrades many coagulation proteins, thus modulating both the thrombotic and the fibrinolytic systems. These interactions should be viewed in a wider context as part of the links between the coagulation and inflammation pathways. PMID- 9347586 TI - Prospects for gene therapy of neutrophil defects. PMID- 9347585 TI - Antimicrobial peptides of phagocytes and epithelia. AB - Human and other vertebrate leukocytes contain multiple distinct antimicrobial (poly)peptides. Of these, BPI is a LPB protein active against gram-negative bacteria, PLA2 specifically cleaves bacterial phospholipids, while defensins and cathelicidins are broad spectrum antimicrobials that preferentially permeabilize microbial membranes. These and other polypeptides function in both phagocytic and extracellular killing of microbes, attacking multiple molecular targets to cooperatively penetrate and disrupt the microbial surfaces and membrane barriers. Such antimicrobial substances are of interest not only for students of phagocytic and epithelial host defenses but may lead to the development of novel pharmaceuticals for the treatment of infections and their sequellae. PMID- 9347587 TI - Projecting Social Security earnings: past is prologue. AB - Accurate projections of lifetime earnings are useful in projecting Social Security benefits, trust fund balances, and economic resources of the elderly and the effects of changes in Social Security policy. This article projects lifetime Social Security earnings until retirement using data from the Survey of Income and Program Participation (SIPP) matched to Social Security records of annual earnings from 1951 through 1993. We first develop, estimate, and test gender specific multiple regression models of 10-year earnings intervals using the matched 1984 SIPP panel. We find strong relationships predicting the mean indexed monthly earnings level in the 10-year period of 1984-93. We then use the models to project (unobserved) Social Security earnings from 1994 through retirement for persons born between 1931 and 1955. By adding projected earnings to observed annual earnings to date, we forecast lifetime Social Security earnings for persons retiring early in the 21st century. PMID- 9347588 TI - Life-cycle aspects of poverty among older women. AB - This article focuses on the relationship between women's economic status earlier in their lives and their poverty status in old age. Previous research on the determinants of poverty among aged women has documented the socioeconomic and demographic correlates of the poor, and has examined the financial impact of adverse later-life events such as widowhood, deterioration of health, and loss of employment. Using data from the National Longitudinal Survey of Mature Women (NLSMW), we find that most women who experience these types of adverse events in their later years do not become poor and that a large majority of older NLSMW respondents who were poor in 1991-92 were poor earlier in their adult lives. Whether women are impoverished by adverse later-life events depends on their economic resources just prior to the event. But, the financial resources available in old age, in turn, depend very much on their long-term economic status throughout much of their adult lives. This article underscores the fact that for most older women, these adverse events do not appear to precipitate poverty spells--at least not within the first couple of years--and directs attention at longer term circumstances that make some women more vulnerable to poverty. PMID- 9347589 TI - Retooling Social Security for the 21st century. AB - Because of the imbalance between promised benefits and available taxes, some reform of Social Security is inevitable. At the same time, perceptions of Social Security are changing rapidly as it moves away from a system where all recipients -whether rich or poor--received more in benefits than they paid in taxes, and where those who were richer consistently received larger net transfers than those who were poorer. Reform is most likely to succeed if it returns to basic principles such as progressivity, equity, and efficiency. Although these principles sometime conflict, they also provide much common ground. For example, if Social Security is meant to meet the greatest needs of the elderly, then increasing the retirement age (which mainly affects the younger and richer elderly) would be preferable to removal of the cost-of-living adjustment (which mainly affects the older and poorer elderly). Efficiency and equity principles, in turn, call attention to some groups--second earners in households, those with few employee tax preferences, those who work many years, and elderly workers- whose net benefits are lower than others who should have less claim to Social Security resources. PMID- 9347590 TI - The Civilian War Benefits program: SSA's first disability program. PMID- 9347591 TI - Pressure ulcer treatment. PMID- 9347592 TI - The effects of surface anaesthesia on the autonomic dysreflexia response during functional electrical stimulation. AB - Recently, increases in blood pressure (BP) and concomitant bradycardia, suggestive of autonomic dysreflexia (AD), have been documented during functional electrical stimulation (FES) in individuals with a high spinal cord injury (SCI). If uncontrolled, this response could preclude the safe use of FES among such individuals. FES induced pain is partly related to stimulation of skin nociceptors. Therefore, measures to reduce skin sensitivity may reduce the risk of AD during FES. The purpose of this study was to determine if topical anaesthetic applied over the site of electrical stimulation could minimize the AD cardiovascular and hormonal responses to FES in individuals with SCI above the T6 level. Seven subjects with a SCI above T6 received FES to the quadriceps muscle of each leg under two conditions on two different testing days. The two treatment conditions, topical anaesthetic and placebo creams, were double blinded and randomized. The cream was administered to an area the size of the electrode (10 x 10 cm) 1 h prior to stimulation. Stimulation began at 0 mAmps and increased by 16 mAmps every 2 min until an intensity of 160 mAmps was achieved. HR and BP were measured at each stimulation intensity level. Catecholamines were analyzed three times during the stimulation protocol (pre, mid and post stimulation intensities). At the end of the stimulation protocol, FES induced isometric quadriceps contraction force at 160 mAmps intensity was measured using a hand held dynamometer. As FES stimulation intensity increased, significant rises in systolic and diastolic BP were seen, with a concomitant progressive drop in HR. The AD response to stimulation was not significantly different between the topical anaesthetic and placebo conditions. Serum catecholamine (epinephrine and norepinephrine) levels tended to rise with increasing FES intensity levels but did not reach statistical significance. The two treatment conditions did not significantly affect serum catecholamine levels or FES-induced quadriceps contraction force. In summary, FES application to the quadriceps muscle in high level SCI subjects resulted in significant increases in BP, decreases in HR (AD like response), a trend towards elevations in catecholamine levels, and no difference in quadriceps muscular strength. However, these responses were unaffected by the use of topical anaesthetic cream on the skin at the stimulation site. This suggests that other mechanisms than skin nociception are operative in FES-induced AD. PMID- 9347593 TI - Self-reported prevalence of pulmonary symptoms in subjects with spinal cord injury. AB - To determine the prevalence of respiratory symptoms in subjects with chronic spinal cord injury (SCI), 180 subjects completed a standard respiratory questionnaire modified for subjects with limited mobility. Subjects were categorized as high tetraplegia (HT:C5 and above not requiring mechanical ventilation), low tetraplegia (LT: C6-8), high paraplegia (HP: T1-7), or low paraplegia (LP: T8-L3). Overall, 68% of subjects reported one or more respiratory symptom. Breathlessness, the most prevalent complaint, was associated with level of lesion: HT = 73%, LT = 58%, HP = 43% and LP = 29%, whereas complaints of cough, phlegm, cough and phlegm, and wheeze did not differ significantly among subjects in the four groups. Breathlessness occurred significantly more often in the group with HT during rest or following exposure to hot air or passive smoke. Awareness of phlegm or wheeze was reported with increased prevalence among subjects with tetraplegia who had complete injuries. Among subjects with tetraplegia, respiratory complaints did not differ significantly in current smokers, former smokers, and non-smokers, whereas among subjects with paraplegia, phlegm and wheeze were reported more frequently, among current smokers. PMID- 9347594 TI - A survey of pain during rehabilitation after acute spinal cord injury. AB - There has been little research on pain in the acute phase of spinal cord injury (SCI) rehabilitation. This study surveyed the pain experience and management strategies in such patients. The subjects consisted of inpatients who were undergoing rehabilitation following their acute injury, and were assessed regarding the presence and type of any pain upon admission to the rehabilitation ward, and reviewed weekly during their stay. They were reassessed on reporting any new pain. Pain intensity was recorded on a Visual Analogue Scale. The maximum intensity of pain during admission was compared to that at discharge. All interventions directed at pain management were documented. Patients were reviewed one year after discharge regarding current pain experience. Almost all of the patients (n = 23; 96%) experienced pain at some stage during their inpatient rehabilitation. Overall pain intensity for those patients with pain during inpatient admission decreased by the time of discharge. At the one year review however, pain intensity tended towards that seen on admission. The reasons for pain tending to increase after discharge were not apparent. Neuropathic and Myofascial Pain Syndrome (MPS) were the most common types of pain experienced. A combination of pharmacological, interventional, physical and psychological approaches were used in pain management. At one year review, neuropathic pain remained common while MPS and orthopaedic pain had decreased. Pain is a common and significant problem for many SCI patients and is a challenge for the treating team to manage. PMID- 9347595 TI - Clinical and magnetic resonance imaging correlation in acute spinal cord injury. AB - The aim of this study was to correlate traumatic spinal cord injury (SCI) patient's outcome with magnetic resonance imaging (MRI) performed within the first 15 days following trauma. We retrospectively analyzed 55 SCI patients. Upon admission, 28 were diagnosed as having a complete SCI (51%), versus 27 with an incomplete SCI (49%). All of the patients with a normal pattern on MRI (four cases), had an incomplete SCI, whereas all patients (15 cases) presenting with a hemorrhage pattern (Type 1) had a complete SCI (P = 0.0001). Fourteen of the 15 individuals (93.4%) with the edema pattern (Type II) had an incomplete SCI (P = 0.001), while the other patient had neurological deterioration, and a syrinx was noted 2 years later (6.6%). Among the 10 individuals showing a contusion pattern (Type III), seven were admitted with an incomplete SCI (70%) and three with a complete SCI (30%). The compression pattern tends to be associated with a complete SCI in 77.8% (seven of nine patients). All patients with a transection pattern on MRI (two cases) were clinically diagnosed as having a complete SCI. Early functional prognosis may be established on the basis of clinical presentation of SCI and associated MRI. Cord hemorrhage and transection are irreversible, while edema has a potential for neurological recovery. Cord contusion tends to be associated with an incomplete SCI, unlike the compression pattern, in which the prognosis depends on the degree of the initial neurological damage. PMID- 9347596 TI - Three-dimensional computed tomography for evaluation of cervical spinal canal enlargement after en bloc open-door laminoplasty. AB - We evaluated the use of three-dimensional (spiral) computed tomography (CT) to assess widening of the bony cervical spinal canal following en bloc open-door C3 C7 laminoplasty in 31 patients. Measurements were performed using a computerised image processing system. The increment in the anteroposterior spinal canal axis and volume of the bony spinal canal was investigated in relation to changes in cervical spine lordosis as well as postoperative neurological improvement. Neurological follow-up averaged 2.6 years (range, 1 to 7 years). A mean increase of 42% was observed in the anteroposterior axis of the canal between C3 to C7 after surgery, while the volume increased by 45%. Postoperatively, cervical spine lordosis correlated with increments in the volume of the enlarged bony canal rather than the anteroposterior axis. Postoperative neurological improvement correlated significantly with the increase in the canal volume. Our results indicate that spiral CT is useful for the assessment of the magnitude of cervical canal enlargement and evaluating the space available for the spinal cord following the relief of compression. PMID- 9347597 TI - Oxygen uptake and heart rate responses during arm vs combined arm/electrically stimulated leg exercise in people with paraplegia. AB - The purpose of this study was to compare the oxygen uptake and heart rate responses during submaximal arm cranking to combined arm cranking + electrical stimulation (ES)-induced leg cycling in individuals with spinal cord injury (SCI). Seven subjects with paraplegia (T4-T12) performed combined arm and leg cycling exercise for 5 min, followed by arm cranking alone at the same power output for a further 5 min. During both exercise conditions, steady state oxygen consumption (VO2), carbon dioxide output (VCO2), expired ventilation (VE) and heart rate (HR) were determined. The respiratory exchange ratio (RER) and oxygen pulse were calculated from the measured variables. During combined arm + electrical stimulation-induced leg cycling exercise, the VO2 was 25% higher (1.58 l min-1 vs 1.26 l min-1), but the HR was 13% lower (132 b min-1 vs 149 b min-1), than during arm cranking exercise alone. Oxygen pulse and VCO2 were also significantly higher (by 42% and 25%, respectively) during combined arm + ES induced leg exercise, but there were no differences between the two exercise conditions for VE or RER. These data suggest that the absence of the leg 'muscle pump' and a reduced venous return of blood to the heart elevate exercise heart rates during submaximal arm cranking. Conversely, combined arm cranking + ES induced leg cycling exercise provides the body with a greater metabolic stress than arm cranking alone, while reducing the cardiac stress. The mechanism explaining the heart rate response, however, remains unclear, but may have been influenced by the blood pressure variations across the range of lesions. The findings from this study may have implications for the relative benefit of combined arm + ES-induced leg cycling training for people with paraplegia. PMID- 9347598 TI - Gabapentin for the treatment of spasticity in patients with spinal cord injury. AB - Our serendipitous observations suggested that some patients with spasticity appeared to have improved following the administration of the anticonvulsant drug gabapentin. As some patients with spasticity are either refractory to or intolerant of established medical treatments, we conducted this study to investigate the effect of gabapentin on spasticity in patients with spinal cord injury. Twenty-five patients with spinal cord injury and spasticity received oral gabapentin (2400 mg over 48 h) in a randomized, double blind, placebo-controlled crossover study. We assessed responses by measuring the Ashworth spasticity scale, muscle stretch reflexes, presence of clonus and reflex response to noxious stimuli. Patient ratings were obtained using a Likert Scale. Administration of gabapentin, but not placebo, was associated with an 11% reduction in spasticity as measured by the Ashworth Scale (P = 0.04) and by a 20% reduction in the Likert Scale (P = 0.0013). Significant changes were not obtained for the other measures. The data obtained suggest that gabapentin may be useful in the management of spasticity associated with spinal cord injury. PMID- 9347599 TI - Treatment of external urethral sphincter hypertonicity by pudendal nerve block using phenol solution in patients with spinal cord injury. AB - We report our experience utilizing the technique of phenol block of the pudendal nerve in the treatment of voiding dysfunction due to hypertonicity of the external urethral sphincter. We have performed 13 pudendal nerve blocks using a 7% phenol solution in seven patients with spinal cord injury who could not obtain relaxation of the external urethral sphincter with a large postvoid urine residual (150 ml to 600 ml) despite large doses of antispasticity drugs and intermittent catheterisations over three weeks. These drugs were discontinued at least 48 hours before this procedure. The efficacy of the pudendal nerve block could also be tested by the ease of facilitating micturition during or just after the block and measuring the amount of postvoid residual urine and intravesical leak pressure. A pudendal nerve block was produced by injecting a 7% phenol solution medial to the ischial tuberosity having specifically localized the nerve by electrical stimulation. This procedure improved the voiding pattern dramatically, leading to a full stream of urine and a remarkable decrease of postvoid residual volume and intravesical leak pressure. The mean difference of the postvoid residual volume and the intravesical leak pressure before and after pudendal nerve block was 255.7 ml and 57.5 cmH2O, respectively. We conclude that pudendal nerve block with a phenol solution as a treatment of external urethral sphincter hypertonicity was effective, easy to perform, and had no complication. This treatment should be considered as a possible alternative to more invasive surgical procedures. PMID- 9347600 TI - Use of pulsed irrigation evacuation in the management of the neuropathic bowel. AB - Management of the neuropathic bowel is one of the major issues in the treatment of patients with severe spinal cord injury (SCI). Pulsed irrigation evacuation (PIE) has been evaluated in several small studies for the clearing of fecal impactions in patients with a neuropathic bowel. We evaluated our experience with 398 PIE procedures performed on inpatients and outpatients at our facility. It has proven to be both safe and effective in a wide variety of patients with this disorder, and is a useful addition to traditional methods in the management of the neuropathic bowel. PMID- 9347601 TI - Intradural neurenteric cyst of the cervical spine misdiagnosed as a psychogenic disorder in a 7-year-old-child. AB - The patient is a 7-year-old Japanese female child with an intradural neurenteric cyst of the cervical spine. She had been misdiagnosed as having a psychogenic disorder due to the absence of neurological changes and bony anomalies. Subjective recovery followed the removal of the cyst. PMID- 9347602 TI - Stage IV Hodgkin's disease presenting with spinal epidural involvement and cauda equina compression as the initial manifestation: case report. AB - Hodgkin's disease very rarely presents clinically, initially with a paraspinal mass, but this should be considered in the differential diagnosis. A patient presenting with back pain was diagnosed to have Stage IV Hodgkin's disease. MRI revealed an extradural and intraspinal soft tissue mass with bone infiltration. The importance of MRI in the early evaluation of a paraspinal mass and in determining the extent of the disease is emphasized. PMID- 9347603 TI - Possible role of denervation-induced changes in the urothelium in the pathophysiology of cystitis in patients with spinal cord injury: a hypothesis. PMID- 9347604 TI - The epiconus syndrome presenting radicular-type neurological features. PMID- 9347605 TI - Multiple thoracic disc herniations--case report and review of the literature. PMID- 9347606 TI - The history of modern spinal traction with particular reference to neural disorders. PMID- 9347607 TI - Synaptic depression: a dynamic regulator of synaptic communication with varied functional roles. PMID- 9347608 TI - Non-invasive optical spectroscopy and imaging of human brain function. AB - Brain activity is associated with changes in optical properties of brain tissue. Optical measurements during brain activation can assess haemoglobin oxygenation, cytochrome-c-oxidase redox state, and two types of changes in light scattering reflecting either membrane potential (fast signal) or cell swelling (slow signal), respectively. In previous studies of exposed brain tissue, optical imaging of brain activity has been achieved at high temporal and microscopical spatial resolution. Now, using near-infrared light that can penetrate biological tissue reasonably well, it has become possible to assess brain activity in human subjects through the intact skull non-invasively. After early studies employing single-site near-infrared spectroscopy, first near-infrared imaging devices are being applied successfully for low-resolution functional brain imaging. Advantages of the optical methods include biochemical specificity, a temporal resolution in the millisecond range, the potential of measuring intracellular and intravascular events simultaneously and the portability of the devices enabling bedside examinations. PMID- 9347609 TI - Luigi Galvani and animal electricity: two centuries after the foundation of electrophysiology. AB - Luigi Galvani and his famous experiments on frogs carried out in the second half of the 18th century belong more to legend than to the history of science. Galvani not only laid the foundations of a new science, electrophysiology, but also opened the way for the invention of the electric battery, and thus for the development of the physical investigations of electricity. However, in spite of the widespread celebration of his work, Galvani's scientific endeavours have been largely misrepresented in the history of science. The scholar of Bologna has a stereotyped image as an 'occasional' scientist, who started his studies by chance, largely ignored the scientific theories of his time and wandered aimlessly in mental elaborations until the physicist of Pavia, Alessandro Volta, entered the field, correctly interpreted Galvani's results and eventually developed the electric battery. With the present understanding of electrical phenomena in excitable membranes, it is now time to reconsider the real matter raised by Galvani's discoveries and by his hypothesis of an intrinsic 'animal electricity', and to make a clearer evaluation of a revolutionary phase of scientific progress. PMID- 9347610 TI - More on calcium currents. PMID- 9347611 TI - A motivational perspective of conscious experience. PMID- 9347612 TI - Network memory. AB - Our thinking on the cortical organization of primate memory is undergoing a copernican change, from a neuropsychology that localizes different memories in different areas to one that views memory as a distributed property of cortical systems. We are shifting our focus from 'systems of memory' to the memory of systems. The same cortical systems that serve us to perceive and move in the world serve us to remember it. Our memories are networks of interconnected cortical neurons, formed by association, that contain our experiences in their connectional structure. Perceptual and motor memory networks are hierarchically organized in post-rolandic and pre-rolandic neocortex, respectively. Recall, recognition and working memory consist largely in their reactivation, also by association. PMID- 9347613 TI - Central clocking. AB - The main questions in circadian neurobiology are: how many oscillators are involved; how are their daily oscillations generated and synchronized to the external world; and how do they signal time of day to the organism. The suprachiasmatic nuclei of the hypothalamus (SCN) are well established as the principal circadian oscillator of mammals. Their 10,000 or so 'clock' neurones drive our overt rhythms-the daily patterning we observe in our physiology and behaviour being mirrored perfectly by their spontaneous cycle of neuronal activity. However, they are not our only circadian oscillator, their molecular timekeeping is not understood and they ways in which they communicate with other parts of the brain are more unusual than was previously assumed. PMID- 9347614 TI - The GluR2 (GluR-B) hypothesis: Ca(2+)-permeable AMPA receptors in neurological disorders. AB - The abnormal influx of Ca2+ through glutamate receptor channels is thought to contribute to the loss of neurons associated with a number of brain disorders. Until recently, the NMDA receptor was the only glutamate receptor known to be Ca(2+)-permeable. It is now well established that AMPA receptors exist not only in Ca(2+)-impermeable but also in Ca(2+)-permeable forms. AMPA receptors are encoded by four genes designated gluR1 (gluR-A) through gluR4 (gluR-D). The presence of the gluR2 subunit renders heteromeric AMPA receptor assemblies Ca(2+) impermeable. Recent studies involving animal models of transient forebrain ischemia and epilepsy show that gluR2 mRNA is downregulated in vulnerable neurons. These observations suggest that downregulation of gluR2 gene expression may serve as a 'molecular switch' leading to the formation of Ca(2+)-permeable AMPA receptors and enhanced toxicity of endogenous glutamate following a neurological insult. PMID- 9347615 TI - Organotypic slice cultures: a technique has come of age. AB - Slices of CNS tissue prepared from young rodents can be maintained in culture for many weeks to months. The basic requirements are simple: a stable substratum, culture medium, sufficient oxygenation and incubation at a temperature of about 36 degrees C. Under these conditions, nerve cells continue to differentiate and to develop a tissue organization that closely resembles that observed in situ. Several alternative culturing methods have been developed recently. Slices maintained in stationary culture with the interface method are ideally suited for questions requiring a three-dimensional structure, whereas slices cultured in roller-tubes remain the method of choice for experiments that require optimal optical conditions. In this report, three typical experiments are discussed that illustrate the potential of the slice-culture technique. The first example indicates that, due to their high neuronal connectivity, slice cultures provide a very useful tool for studying the properties of synaptic transmission between monosynaptically coupled cell pairs. The other two studies show how long-term application of substances to slice cultures can be used to examine the consequences of epileptic discharges in vitro, as well as the effects of slowly acting clostridial neurotoxins on synaptic transmission. PMID- 9347616 TI - Neural transplantation studies reveal the brain's capacity for continuous reconstruction. AB - Basic research using cell transplantation indicates that structural developmental mechanisms seen in immature brains can also function in the adult brain. As the brain matures, cellular migration and axonal growth is impeded. However, fetal neural transplantation studies have shown that directional cues are available for fetal axons to find specific host neurons in the adult brain. By reaching specific and distant CNS target zones, donor tissue with extended axonal growth periods demonstrate both an abundance and specificity of CNS neurotropic signals. The presence of specific guidance cues, despite strong inhibition of regenerative long-distance axonal growth, suggests that these cues play other physiological roles in the adult CNS, and could be utilized therapeutically for reconnection of neuronal pathways. PMID- 9347617 TI - Adenosine-dopamine receptor-receptor interactions as an integrative mechanism in the basal ganglia. AB - Increasing evidence suggests that antagonistic interactions between specific subtypes of adenosine and dopamine receptors in the basal ganglia are involved in the motor depressant effects of adenosine receptor agonists and the motor stimulant effects of adenosine receptor antagonists, such as caffeine. The GABAergic striatopallidal neurons are regulated by interacting adenosine A2A and dopamine D2 receptors. On the other hand, the GABAergic striatonigral and striatoentopeduncular neurons seem to be regulated by interacting adenosine A1 and dopamine D1 receptors. Furthermore, behavioural studies have revealed interactions between adenosine A2A and dopamine D1 receptors that occur at the network level. These adenosine-dopamine receptor-receptor interactions might offer new therapeutic leads for basal ganglia disorders. PMID- 9347618 TI - Rhythmic transcription: the molecular basis of circadian melatonin synthesis. AB - Adaptation to a changing environment is an essential feature of physiological regulation. The day-night rhythm is translated into hormonal oscillations governing the metabolism of all living organisms. In mammals the pineal gland is responsible for the synthesis of the hormone melatonin in response to signals originating from the endogenous clock located in the hypothalamic suprachiasmatic nucleus (SCN). The molecular mechanisms involved in rhythmic synthesis of melatonin involve the cAMP response element modulator (crem) gene, which encodes transcription factors responsive to activation of the cAMP signalling pathway. The CREM product, inducible cAMP early repressor (ICER), is rhythmically expressed and participates in a transcriptional autoregulatory loop that also controls the amplitude of oscillations of 5-HT N-acetyl transferase, the rate limiting enzyme of melatonin synthesis. Thus, a transcription factor modulates the oscillatory levels of a hormone. PMID- 9347619 TI - Changes of tissue-specific transcription factors in the rabbit mammary gland during pregnancy and lactation. AB - In the mammary gland four tissue-specific transcription factors have been found involved in the regulation of milk protein genes: mammary gland-specific nuclear factor (MGF), milk protein binding factor (MPBF), pregnancy-specific mammary nuclear factor (PMF), mammary cell-activating factor (MAF). Rabbit beta-casein gene promoter contains motifs highly homologous (89-100%) to MGF, MAF and PMF consensus sequences. These transcription factors were analysed simultaneously using electrophoretic mobility shift assays (EMSA) on nuclear protein extracts derived from pregnant or lactating rabbit mammary glands. The specificity of these complexes was analysed in cross-competition EMSA experiments. It was shown that in the rabbit mammary gland the DNA-binding activities of MGF, MAF and PMF change during pregnancy and lactation. Protein extracts of nuclei isolated from mammary glands of 15-day pregnant rabbits form fast-migrating low-specific complexes with MGF, MAF and PMF oligonucleotide probes; in the extracts from mammary glands of late pregnant (day 25) and lactating (day 5) rabbits additional slowly-migrating highly specific DNA-protein complexes are formed. Their appearance changes in parallel with the activation of beta-casein gene expression as measured by run-on gene transcription and beta-casein mRNA accumulation. PMID- 9347621 TI - Development of a slide ELISA for canine leishmaniasis and comparison with four serological tests. AB - A slide ELISA for canine leishmaniasis was developed by using promastigotes of Leishmania infantum, and compared with microimmunodiffusion, immunoelectrophoresis, direct agglutination and indirect immunofluorescence assays. The sensitivity of all the tests was 100 per cent. The specificity of the direct agglutination test was 95 per cent but it was 100 per cent for the three other tests. There was also a positive correlation and a high level of concordance between the titres measured by the different tests. PMID- 9347620 TI - The origins of benevolence in the veterinary profession. PMID- 9347622 TI - Ultrasonographic findings in cows with left displacement of the abomasum. AB - Forty-eight cows with left displacement of the abomasum (LDA) and three clinically healthy control cows were examined ultrasonographically from the 11th and 12th intercostal spaces on the left side. In the controls, the rumen was immediately adjacent to the left abdominal wall, whereas in the cows with LDA the rumen was generally immediately adjacent to the left abdominal wall ventrally, but displaced by the abomasum more dorsally. The ultrasonographic findings were generally consistent in the cows with LDA. The ingesta that were visualised ventrally in the abomasum appeared echogenic to hypoechogenic and, in a few cows, the abomasal folds were visible as elongated, echogenic, sickle-shaped structures. The dorsal abomasal gas cap was characterised by reverberation artifacts from the abomasal surface. PMID- 9347623 TI - Expression of non-cytopathogenic bovine viral diarrhoea virus (BVDV) in oocytes and follicles of persistently infected cattle. PMID- 9347624 TI - Caries and odontoclastic resorptive lesions in a chinchilla (Chinchilla lanigera). PMID- 9347625 TI - Concurrent infection with enteric protozoa and Erysipelothrix rhusiopathiae in chicken and pheasant flocks. PMID- 9347626 TI - Survey on lungworm in adult cattle. PMID- 9347627 TI - Sensitivity of the single intradermal comparative tuberculin test. PMID- 9347628 TI - Animal experiments. PMID- 9347629 TI - Preventive role in cattle production. PMID- 9347630 TI - ECG of the month. Name that block! Trifascicular block. PMID- 9347631 TI - Cerumen impaction. AB - Cerumen impaction represents the most common otologic problem encountered by physicians. It can affect up to 6% of the general population and a much higher percentage in the mentally retarded population. Cerumen is a mixture of secretory products of two glands in the external auditory canal where it serves a protective function. The external auditory canal possesses a unique anatomy and physiology that permits an efficient self-cleaning system. In most cases, a breakdown in the epithelial migration of the external auditory canal is thought to cause cerumen impaction. The management of cerumen impaction include direct removal, irrigation, and the use of cerumenolytics. PMID- 9347632 TI - Radiology case of the month. Delayed treatment of a large, fungating mass. Benign giant-cell tumor of bone. PMID- 9347633 TI - The Journal 100 & 150 years ago. Medicine 100 & 150 years ago. October 1847 & 1897. PMID- 9347634 TI - Minor advocacy in healthcare: a proposal for a more efficient solution. AB - Autonomy is a crucial, yet litigious right in health care that is usually unquestioned in the case of adults. However, children's or their guardian's refusal to accept necessary medical care can expedite the question of autonomy from the medical to the legal arena. Therefore, to save time, money, and lives, the health care establishment, specifically ethics committees, must be given rights as a judicial body to hand down binding judgments in cases of minors refusing the medical community's standard of care. PMID- 9347635 TI - Which parishes are most healthy? AB - Raw data for 21 health outcomes are ranked from 1 to 64 within each measure for all Louisiana parishes. The average of all ranked outcome measures is reported along with the individual ranked scores. This average rank for all parishes is also reported as a quartile score. The ranked measures are grouped in eight categories suggested by the Healthy People 2000 program, and a quartile score is reported for the average of each of the categories. The purpose of this rankings is to enable comparisons of health outcomes between and within the parishes. The diversity of the measures within parishes suggests strengths and weaknesses for each community. As suggested by the Healthy People 2000 program, health outcomes, when organized in the conceptual categories, suggest how behavior and attitude impact on health status. PMID- 9347636 TI - Free flap breast reconstruction: the LSU experience (1984-1996). AB - From 1984 through 1996 the section of Plastic and Reconstructive Surgery at Louisiana State Medical Center has performed over 330 breast reconstructive procedures with free flaps. Seven types of reconstructive procedures have been used during this time span, each with its specific salient positive and negative points. The breast reconstruction techniques included the use of (1) Superior Gluteal Myocutaneous Free Flap, (2) Superficial Inferior Epigastric Artery Flap, (3) Transverse Rectus Abdominis Myocutaneous Free Flap, (4) Deep Inferior Epigastric Perforator Flap, (5) Superior Gluteal Artery Perforator Flap, (6) Inferior Gluteal Artery Perforator Flap, and (7) Lateral Thigh Perforator Flap. The experience with these different methods of breast reconstruction has led us to believe that the ideal material for breast reconstruction is skin and fat, rather than muscle or prosthetic devices. At our institution we have evolved from the myocutaneous flap to the use of perforator flaps for breast reconstruction: the donor site morbidity is less, the 99% success rate is superior, and it allows more options with the perforator free flaps than ever realized with the myocutaneous free flap technique. We feel that, in the future, these perforator techniques will become the standard for autogenous breast reconstruction. PMID- 9347637 TI - Anti-E as a cause of hemolytic disease of the newborn. AB - A case of HDN caused by anti-E antibody is reported. A group A, E-positive, hemoglobin E trait female infant was born from a group A, E-negative, beta thalassemia/hemoglobin E mother. Hyperbilirubinemia was noted at the first day of life. The DAT was positive. Anti-E was detected in the maternal serum. Jaundice and anemia occurring to the baby were severe enough to require phototherapy intervention for 9 days and 50 ml of group A, E-negative packed red blood cells was transfused. The baby's condition improved. She was discharged at 12 days of age. Follow-up of the baby at 1 year old showed that she was alive and in good health. PMID- 9347638 TI - A comparative study of three techniques for eluting red cell antibodies. AB - Sixty-seven eluates obtained from the heat, ether and acid elution techniques were tested with the specific red blood cells (RBCs) and were compared according to their reactivities using the indirect antiglobulin test (IAT). It was found that the ether elution technique was superior in eluting Rh antibodies except for anti-e while the acid elution technique was superior in eluting Miltenberger (Mi(a)) antibodies (P < 0.05). The heat elution technique gave the lowest reactivity among the three techniques. In conclusion, the reactivities of the eluates obtained from the acid elution technique were overall comparable to those from the ether elution technique. The acid elution technique is practical for routine use in most blood banks because it is less time consuming and reduces the risk of exposing hazardous chemicals. PMID- 9347639 TI - Effectiveness of leukocyte removal by Imugard III. AB - Study was conducted to evaluate the effectiveness of Imugard III-RC (4B) to remove leukocyte from packed red cells (PRCs). The leukafiltration set, Imugard III (4B), bedside use for PRCs or whole blood was converted to laboratory use by connecting the distal end of filter tubing to the transfer bag and the proximal end of tubing to the primary blood pack by sterile connecting device to ensure that the closed system is intact. Twenty units of PRCs were prepared from CPD whole blood by removal of platelet rich plasma. All units were stored at 4 degrees C for 1 day before filtration. Volumes of PRC/Bag ranged from 240-340 ml, mean = 276.75 ml and SD = 26.57. Prefiltration WBC/Bag ranged from 464 x 10(6) 5910 x 10(6) cells, mean = 2862.05 x 10(6) cells and SD = 1280.87 x 10(6). The filtration time/Bag ranged from 18-45 min, mean = 27.3 min and SD = 7.9. The WBC removals ranged from 99.99 per cent-100 per cent, mean = 99.99 per cent and SD = 0. The residual WBC in PRC/Bag ranged from 0-0.24 x 10(6) cells, mean = 0.134 x 10(6) cells and SD = 0.063 x 10(6). The RBC loss from PRCs with no additive solution was approximately 15 per cent. There was no need for priming and purging the filtration set with saline before and after filtration. CONCLUSION: The reduction of WBC by 20 sets of Imugard III-RC (4B) was 99.99 per cent or higher with the residual WBC < 1 x 10(6) which is able to reduce nonhemolytic transfusion reaction, CMV transmission and HLA alloimmunization. PMID- 9347640 TI - DNA typing of the HLA-A, -B and -C genes: possible MHC class I haplotypes in the northeastern-Thais. AB - The phenotype and gene frequencies of HLA class I were studied in the Northeastern Thai population. Blood samples were collected from 100 unrelated healthy northeastern-Thais. HLA-A, -B and -Cw alleles were determined using the polymerase chain reaction- amplification refractory mutation system (PCR-ARMS). 12 HLA-A, 20 HLA-B and 14 HLA-Cw alleles were found. Linkage disequilibrium analysis indicated the existence of 7 HLA-A-B and 19 HLA-B-Cw haplotypes. A*0207 Cw*01-B*4601 was the most common possible haplotype in this population. These results provide regional basic information for further studies in anthropology, organ transplantation and MHC disease associations in the northeastern-Thais. PMID- 9347641 TI - HLA class II allele frequencies in northern Thais (Kamphaeng Phet). AB - The polymorphism of HLA class II genes (HLA-DRB1, -DQA1, -DQB1 and -DPB1) was investigated in 97 normal Northern Thais (NT) from Kamphaeng Phet province using PCR-SSO typing. Allele frequencies (AF) have been determined. DRB1*1202 (17.5%), DRB1*1502 (16.5%), DQA1*0101 (25.8%), DQA1*0102 (21.7%), DQB1*0502 (22.7%), DPB1*0501 (23.2%) and DPB1*1301 (22.7%) showed the highest frequencies in each locus. These results were more similar to those observed in Present-day Thais (PDT) and Central Thais (CT) than Northern Thais from Chiang Mai (CM) and Dai Lue (DL). However, the data presented in this population study should be useful in many fields, such as anthropology, organ transplantation, disease susceptibility and evolutionary genetics. PMID- 9347642 TI - A*02 in southern Thai Muslims and central Thais. AB - The HLA-A*02 subtyping in Thais was conducted and included in the 12th International Histocompatibility Workshop (12WS). A total of 81 randomized individuals previously serologically or DNA typed as A2 were studied for A2 subtypings. The subjects consisted of 32 Southern Thai-Muslims (STM) and 49 Central Thais (CT). The 12WS HLA-A*02 subtyping DNA typing kit was employed. The most common A*02 subtypes in STM were A*0203,*0201 and *0207 while they were A*0203, *0207 and *0201 in CT. A*0202, *0204, *0208, *0209, *0212, *0213, *0214, *0215, *0216 and *0217 were not found in both STM and CT. The 12WS data indicated that A*0201 was also the most frequent allele of A*2 among North-East Asians. A2 subtype study in 32 STM revealed that 2 in 8 of A*0201 showed the absence of bands at 813 bp and 705 bp with primer mix number 03A and 517A and weak reaction band with primer mix number 33A. In addition, 3 subjects with A*0201 variations have one nucleotide difference in exon 2 by sequence base typing (by MGJ. Tilanus) which will be reported separately. CONCLUSION: More variations of A*02 were observed among STM compared to CT. The variations of reactions with the set of primer mix should be carefully observed and subjected to further analysis. PMID- 9347644 TI - HLA gene frequencies of northern Thais. AB - We investigated the distribution of HLA-A and B locus, gene frequency (GF), antigen frequency (AF), haplotype frequency (HF) and non detectable antigens in Northern Thais. Of 289 native northern Thai people residing in Chiang Mai province for many generations were tested using lymphocytotoxicity test and 146 unrelated subjects were selected for analysis. The common alleles were A2, A11 and A24 for A locus with GF of 36.4%, 35.4% and 15.6%, respectively and B46, B40 and B13 for B locus with GF of 21.1%, 15.7% and 8.6%, respectively. The frequent linkage disequilibrium haplotypes were A2,B46; A33,B17 with HF of 15.9%; 5.0% and LD of 8.3%; 4.6%, respectively (p < 0.0001). The undetectable antigens (blanks) occurred with GF = 11.64% at A locus and GF = 4.92% at B locus. Comparing the GFs to other Thai ethnic groups, showed that the Northern Thais shared several alleles such as A2, A11, B46, and B62 in common with Dai Lue (Thai-speaking people who lived in the southern part of China), (p > 0.05), more than Thais, Thai/Chinese or present-day Thais (p < 0.001). Especially, HLA-B46 with the GF of 21.1% is considered to be a very typical antigen for Southern Mongoloids. These similarities will support the root of migration and origin of Northern Thais. PMID- 9347643 TI - HLA in southern Thai-Muslims. AB - One hundred and two Southern Thai-Muslims (STM) from Nakhon Si Thammarat province were studied for HLA class I and II by SSP ARMS-PCR and PCR-SSO, respectively. The allele frequencies, haplotype frequencies, delta value and linkage disequilibrium between alleles were expressed. The most frequent alleles for HLA A, HLA-B and HLA-C were A*24(02,03), A*11 (01,02), A*02(01,03,05-07,11): B*15(01,04-07,12,19,20), B*07(02-05), B*51(01-05)/B*52 (011,012); and Cw*07(01 03), Cw*04(01,02), Cw*08(01-03), respectively. The HLA class II alleles frequently found were DRB1*1202, DRB1*15021, DRB1*0701; DRB3*0301; DRB5* 0101; DQA1*0101, DQA1*0103, DQA1*0601; DQB1*0301, DQB1*0501, DQB1*0201; and DPB1*1301, DPB1*2301 and DPB1*0501. Two common HLA class I and II haplotypes with significant linkage disequilibrium were A*24 (02,03)-Cw*08 (01-03)-B*15 (01,04 07,12,19,20) -DRB1*1202 and A*33 (01,02)-Cw*0302-B*5801-DQB1*0201. The absence of B*27 and DRB1 *1401, the presence of A*2301 and high frequency of A*68 were observed in STM. CONCLUSION: Certain level of genetically distinction among STM, CT and NET existed. However, the genetic diversity of STM was relatively closer to CT than NET. PMID- 9347645 TI - Increasing the chance to accept HLA-ABDR mismatched donor in kidney transplantation. AB - Two hundred and fifty-three kidney transplantations (KT) which included 68 (26.9%) living-related (L) and 185 (73.1%) cadaveric (C) KT with 0-6 HLA-ABDR mismatches (MM) were studied for the association of HLA-ABDR-MM specificities and the occurrence of graft rejection (GR). It was found that the incidence of acute and chronic rejection in CKT was significantly higher than that of LKT (42.1% vs 22.1%, p < 0.005). It was also observed that the number of ABDR-MM, AB-MM and BDR MM which is important in GR were 2 times in CKT compared with LKT. The analysis revealed that HLA-A11, B16, B22, B35, B5, B17 and DR3 were good responders, whereas, HLA-A30, A2, B62, B18, B40, B44, B46 and DR10 were good stimulators for KT. GR were significantly increased with p < 0.01 and < 0.05, respectively. Specific HLA-MM specificities played a significant role in GR, i.e., some HLA-MM specificities were permissible, whereas, some were immunogenic. Careful selection of donor and recipient for KT by avoiding immunogenic HLA-MM and/or accepting permissible HLA-MM will improve graft survival and reduce the demand of kidney for retransplantation. PMID- 9347646 TI - Panel reactive antibodies in post-kidney transplantation. AB - This study was aimed to evaluate the clinical relevance of the panel reactive antibodies (PRA) post kidney transplantation (KT). A total number of 90 KT recipients consisted of 71 male and 19 female patients. Thirty-two haploidentical and 3 HLA-identical pairs for living related KT and 55 cadaveric KT with 3-6 mismatched antigens were included in this study. The analysis revealed that there were 2 out of 69 (2.89%) patients with no episode of rejection who had Pre-KT PRA T and or PRA-B > 80 per cent while they were 5.79 per cent and 23.19 per cent for Post-KT. No patient in 21 cases with KT rejection had Pre-KT-PRA-T and -B > 80 per cent. There was significant increase of antibodies in Post-KT rejections which were 28.57 per cent and 33.3 per cent for Post-KT-PRA-T and -B respectively. None of 3 cases with graft failure (GF) from chronic rejection had Pre-KT-PRA-T and -B > 20 per cent and only one of them had Post-KT-PRA-T = 80 per cent. No donor specific HLA antibody was found among this group of patients. Although antibody to donor HLA antigens was not observed in these patients, the increase of PRA-T and -B in Post-KT may indicate the immunological reaction resulting in GF. PMID- 9347647 TI - Crossmatching technique facilitating kidney transplantation. AB - Accelerated acute cellular rejection (AR) continues to be a serious problem in kidney transplantation (KT), suggesting that undetected presensitization may be encountered. The purpose of this study was to determine the most sensitive crossmatching (XM) technique to detect the preformed antibody (Ab) which may cause AR. One hundred and twenty two sera from 98 patients, on the waiting list for KT at Ramathibodi Hospital were XMed with 23 cadaveric splenic lymphocytes including 2 living related KT (LR-KT). The XM was performed by 3 different techniques namely, standard microlymphocytotoxicity test (standard NIH), antihuman globulin microlymphocytotoxicity test (AHG) and flow cytometric XM (FCXM). The XM results revealed that 8 out of 75 (10.7%) tests were negative by standard NIH, i.e., 5 tests were positive by AHG only and 1 test was positive by FCXM only and 2 tests were positive by both AHG and FCXM. In addition, the patients who had the AHG technique were not done, 5 out of 47 (10.7%) tests were also negative by standard NIH but were positive by FCXM. The sensitivity of the techniques was done by titrations of anti HLA-A2. It was found that FCXM was the most sensitive technique, followed by AHG and standard NIH, consecutively. In the retrospective study of LR-KT, case #1, the standard NIH for XM using pre-KT blood sample was negative while AHG and FCXM were strongly positive. The patient had AR at day 2 post-KT which confirmed by needle biopsy. The serum at day 11 and day 116 post-KT were tested again and were positive by the 3 techniques. Case #2, pre KT blood sample showed negative T-XM by the 3 techniques while auto-B and B-XM were positive by standard NIH and AHG but negative by FCXM. This patient had rejection at day 16 after KT. The post-KT blood sample at day 30 showed positive auto T/B and T/B-XM by standard NIH and AHG whereas it was still negative by FCXM. It was also noted that Ab to donor B cell was better detected by standard NIH and AHG than FCXM. In conclusion, FCXM is more sensitive than standard NIH and AHG, however this technique is limited in detecting IgM T and B cell Ab. AHG technique can detect both IgG and IgM antidonor T and B cell Abs. In addition, AHG technique is more sensitive than standard NIH and does not require sophisticated equipment. AHG technique should be appropriate for routine XM, especially, in LR-KT and sensitized patients. PMID- 9347648 TI - Evaluation of the reticulocyte count portion of technicon H*3 blood analyzer: lowering the test expense by reducing the reticulocyte reagent. AB - Automated reticulocyte counting has become an essential instrument of the hematology laboratory. This automatic technique has lead to diminishing labour tasks and to significant improvements in accuracy and precision compared with the manual microscopic methods. In any event, it adds a considerable expense to the laboratory budget. Here, we report the modified method of applying the new mixture of 1 microL of whole blood with 1 mL of reticulocyte reagent, which we evaluated for its accuracy and precision, instead of using the mixture of 3 microL of whole blood with 3 mL of reticulocyte reagent recommended by the company. We demonstrated the accepted accurate and precise results of percentage and absolute number of retculocyte count, low-stained reticulocyte count and its corpuscular indices; the mean reticulocyte corpuscular volume (MCVr), mean reticulocyte corpuscular hemoglobin concentration (CHCMr), and mean reticulocyte hemoglobin content (CHr). These suggested that, for every red cell assessed, the number, the cell volume, hemoglobin content and concentration are accurately and precisely measured by the modified method while the sub-populations of reticulocyte count and distribution width of reticulocyte indices are variable. In conclusion, our results provided the information that 1) the modified method can be used as a routine test and it provides accurate and precise results; 2) with the modified method, two-thirds of the expense spent for reticulocyte reagent can be saved; 3) it should not be used for research purposes. PMID- 9347649 TI - Levels of serum tumor necrosis factor alpha in relation to clinical involvement and treatment among Thai adults with Plasmodium falciparum malaria. AB - Concentrations of tumor necrosis factor alpha (TNF-alpha) in serum were measured in 17 Thai men infected with Plasmodium falciparum malarial infections to determine whether they were affected by severity of infections or exchange transfusions. Twelve patients were considered having complicated malarial infections, eight of whom had cerebral malaria. Five patients had uncomplicated malarial infections. The results showed that malarial infection markedly raised TNF-alpha level above normal values (mean +/- SEM 406 +/- 38 vs 15 +/- 5, p = 0.004). In complicated malaria, cerebral involvement appeared to significantly increase concentration of TNF-alpha when compared to values in uncomplicated malaria (mean +/- SEM 496 +/- 64 vs 339 +/- 12, p = 0.01). Degree of parasitemia, intravenous quinine (day 0 value vs day 7 value) and exchange transfusion did not significantly affect TNF-alpha levels. CONCLUSION: Serum level of TNF-alpha is increased in Plasmodium falciparum malarial infections and may be a useful index to predict severity of malarial infection, cerebral malaria in particular. PMID- 9347650 TI - Hematologic parameters in Thai subjects over 50 years old. AB - Advancing age is associated with the decline of hemoglobin, red blood cell count and other parameters. The conventional normal values of hematologic parameters established by all age populations may not be suitable for the elderly. This study determined the hematologic parameters of Thai subjects aged over 50 years by using a fully automated cell counter. No significant age-related variations of all parameters were found. The MCV of both sexes showed an upward trend at age over 70. The red blood cell count and hemoglobin levels in males were higher than females. These, however; were not significant alterations. These hematologic values should be useful for screening and clinical investigation of the elderly. PMID- 9347651 TI - The effect of standardized ginseng extract on peripheral blood leukocytes and lymphocyte subsets: a preliminary study in young health adults. AB - The present study was undertaken in twenty young, healthy Thai males to investigate the effects of prolonged administration of standardized ginseng extract on peripheral blood leukocytes and lymphocyte subsets. The subjects were divided into two equal groups as ginseng and placebo groups. The first group received two capsules daily of standardized ginseng extract 150 mg per capsule for 8 weeks. The second group received placebo and served as control. Circulatory levels of total and differential leukocyte counts and percentage of lymphocyte subsets were determined prior to and at the 4th and 8th week of the experimental period. There were no significant differences in the total and differential leukocyte counts as well as the lymphocyte subpopulations: CD3 (T cells), CD 19 (B cells), CD4 (T-helper cells), CD8 (T-suppression cells), CD4/CD8 ratio, and CD25 (Interleukin-2-receptor cells) between the two subject groups throughout the experimental period. We concluded that oral administration of standardized ginseng extract, 300 mg/day for 8 weeks caused no significant changes in peripheral blood leukocytes and lymphocyte subsets in young, healthy Thai males. PMID- 9347652 TI - Levels of serum interleukin-6 and tumor necrosis factor in postsplenectomized thalassemic patients. AB - Levels of serum interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) were studied in 34 nonsplenectomized thalassemic patients (Thal/nonsplenec), 43 postsplenectomized thalassemic patients (Thal/postsplenec), 13 splenectomized non thalassemic patients (nonThal/ postsplenec) and 18 normal control by enzyme linked immunosorbent assay method. Serum IL-6 concentration in Thal/postsplenec was significantly increased when compared with Thal/ nonsplenec and normal volunteers (3.55 +/- 2.47 pg/ml vs 2.38 +/- 2.31 pg/ml, p = 0.036 and 3.55 +/- 2.47 pg/ml vs 2.66 +/- 0.45 pg/ml, p = 0.028, respectively). This study also demonstrated that TNF-alpha value in Thal/postsplenec was drastically increased above normal control level (15.8 +/- 4.86 pg/ml vs 9.16 +/- 2.18 pg/ml, p = 0.001) and the level was statistically significantly higher than that in Thal/ nonsplenec (15.5 +/- 4.86 pg/ml vs 9.96 +/- 5.19 pg/ml, p = 0.001). There was a trend toward increasing of cytokine levels in Thal/postsplenec with higher platelet count although no correlation was observed. This study addresses the possible role of IL-6 and TNF-alpha in the pathogenesis of reactive thrombocytosis in Thal/postsplenec. PMID- 9347654 TI - Eluding transfusion-transmitted HIV infection: report of two cases. AB - Two hemophilia A boys (FVIII: C < 1% and 2.2%), whose ages were 12 and 14 years. old, received fresh frozen plasma of 140 ml and 210 ml, respectively, in 1989. It was the 27th and 13th donation for each regular donor who was negative for anti HIV testing. However, both donors had HIV seroconversion within 95 to 110 days after the last donation. They might have contracted HIV infection shortly after the last donation. Luckily, the two hemophiliac recipients are still in good health and negative for anti-HIV and HIV-antigen testings for 7 years. PMID- 9347653 TI - Control cells for both electrical impedance and light scattering automated hematological analyzers: preparation from normal and thalassemic blood samples. AB - The study on quality control of automated blood cell analyzers, Technicon H*1 and Coulter MAXM by using three separately self-prepared control cells was extensively investigated. The three parts of control cells are pseudo-leukocyte and fixed platelets, which are fixed by glutaraldehyde, and control red cells from normal and thalassemic patients preserved and anticoagulated in CPD or CPDA 1. The Technicon H*1 system was based on the principle of light scattering but the Coulter MAXM was based on the principle of electrical impedance for cell counting and measurement. The self-prepared control cells can be satisfactorily utilized as control for each system with statistically significant difference (p < 0.05) for both systems. The expired dates for control cells are different in both systems and should be determined for each system specifically. The control red cells prepared from thalassemic patients were quite satisfactorily useful as an abnormal control for both systems during this study. PMID- 9347655 TI - Seroepidemiology of HTLV-I infection in northeast Thailand: a four year surveillance. AB - The human T-lymphotropic virus type I (HTLV-I) can be transmitted through blood transfusion, sexual contact, perinataly and by breast feeding. We carried out a four years seroepidemiology surveillance study of HTLV-I infection among northeast Thai population by screening for antibodies to HTLV-I (anti-HTLV-I) in 1992, 1993, 1995 and 1997. A total of 8,323 blood samples were collected from 6,228 blood donors, 832 pregnancies, 219 multitransfused patients, 53 HIV positive intravenous drug users and 1,000 northeast-Thai workers at different periods of time. The serum samples were tested for anti-HTLV-I by particle agglutination (PA) technique and confirmed by Western blot. One sample from a multitransfused patient collected in 1992 and one sample from a blood donor collected in 1995 demonstrated positive anti-HTLV-I screening by PA but negative by Western blot. This finding indicates that at present HTLV-I is not a public health problem in the northeast of Thailand but surveillance should be continually conducted. PMID- 9347656 TI - Association of HCV and Treponema pallidum infection in HIV infected northeastern Thai male blood donors. AB - The study was performed to determine the association of seroprevalence of hepatitis C virus (HCV) and Treponema pallidum (T. pallidum) infection among HIV infected first time male blood donors (HIV group) in comparison with the HIV seronegative blood donors (control group) in the Northeast of Thailand (NET). Serum samples were collected from 10,321 first blood donation voluntary male donors. All samples were screened for anti-HIV and anti-HCV by particle agglutination test, and syphilis antibody by RPR. The anti-HIV positive sera were repeated by EIA and confirmed by western blot. The reactive anti-HCV samples were confirmed by EIA whereas reactive syphilis antibody samples were confirmed by TPPA. Fisher's exact test was used for statistical analysis. The prevalence of anti-HIV in first time male donors was 0.70 per cent (72/10,321). The age of HIV group and 10,018 male control group ranged from 17-50 years old. The prevalence of HIV among 21-40 years old age group was significantly higher than the 17-20 years old (p = 0.00003). The 17-20 years old HIV group showed significantly higher sero-prevalence of TPPA (p = 0.003). The 21-30 years old HIV group gave significantly higher sero-prevalence of anti-HCV (p = 0.0008) and TPPA (p = 0.045), but the seroprevalence of anti-HCV and TPPA among the 31-50 year old group were nonsignificantly different (p > 0.05). The concurrence of anti-HCV and TPPA in HIV groups was not found. This result indicated that HIV infection among NET voluntary male blood donors was significantly associated with T. pallidum infection in young adults and the HCV infection in mature adults. PMID- 9347657 TI - Detection of HIV-1 window period infection in blood donors using borderline anti HIV results, HIV-1 proviral DNA PCR, and HIV-1 antigen test. AB - Prevention of transmission of HIV-1 via blood transfusion has been carried out by the National Blood Center by screening donated blood with anti-HIV and HIV antigen tests. To increase the safety measure, detection of proviral DNA by PCR has been proposed; however, it was impractical to test all samples by PCR. From August 1994 to September 1995, there were 296,169 blood donors with 0.32 per cent prevalence of anti-HIV positive. From these donors, 153 samples of which the anti HIV enzyme immunoassay optical density (OD) between cutoff and 80 per cent of cutoff value (borderline results) were selected for PCR testing. One out of 153 borderline cases showed positive by PCR test for HIV-1 proviral DNA. However, this case was also positive by HIV antigen test. Therefore, most of the samples with borderline anti-HIV results were true negative for HIV infection. On the other hand, there were 8 HIV antigen positive samples which had anti-HIV OD below the borderline value determined in this study. This finding confirmed the necessity of using both the anti-HIV and HIV antigen tests for screening of donated blood. PMID- 9347658 TI - Total antioxidant capacity in plasma of HIV-infected patients. AB - Total antioxidant capacity (TAC) of fasting EDTA plasma of 33 healthy and 64 HIV infected patients was determined using H(2)O(2)-peroxidase-ABTS technique. The results revealed that the average TAC in HIV-infected patients was significantly lower than those in healthy normal persons. (0.161 +/- 0.097 vs 0.269 +/- 0.081 mmol/L Trolox equivalent, p < 0.05). Total lymphocytes were also counted using Hycel automatic cell counter and absolute CD4 numbers using Coulter CD4 manual kit. It was interesting that CD4 count was not correlated with the clinical symptoms of the patients. This paper suggests that prediction of severity and monitoring of the disease should be performed by determining both total lymphocyte count and total antioxidant capacity. PMID- 9347660 TI - Production and evaluation of Taq DNA polymerase. AB - Taq DNA polymerase is an enzyme essential in performing Polymerase Chain Reaction (PCR) which has recently become a basic technology in research and diagnostic laboratories. In order to reduce the cost of research work in Thailand, recombinant Taq DNA polymerase was locally produced from pTaq cloned in E. coli. The enzyme was characterized and evaluated in comparison with the commercial Taq DNA polymerase produced by Perkin Elmer Cetus, U.S.A. The yield of enzyme was 6.72 mg/ml and the activity of 9,524 units/mg protein with the total of 448,000 units/litre of the bacterial culture. The preparation was free of DNase based upon its ability to degrade Lambda DNA evaluated by gel electrophoresis. Although the enzyme produced gave a high DNA polymerase activity, the preparation was not as pure as the enzyme produced by Perkin Elmer Cetus. Immunoblot analysis indicated that the enzyme preparation contained the products of enzyme degradation obtained during preparation and bacterial protein contaminations. In spite of the existence of bacterial proteins in the preparation, the Taq enzyme produced was proved to be applicable in performing PCR such as the PCR-SSP (Sequence Specific Primers) typing for HLA-DR. The cost of enzyme preparation was about 256 times less than that of the commercial enzyme. Economically, the locally produced Taq DNA polymerase can be used efficiently in the research laboratories performing PCR based typing of the HLA genes. PMID- 9347659 TI - Lack of mutagenicity of stevioside and steviol in Salmonella typhimurium TA 98 and TA 100. AB - Stevioside, a sweet-tasting diterpene glycoside derived from Stevia rebaudiana, and steviol, a product from enzymatic hydrolysis of stevioside, were tested for mutagenic activity by the in vitro Ames test, a preincubation method, using Salmonella typhimurium TA 98 and TA 100 as the tester strains, either in the presence or absence of metabolic activating system derived from the sodium phenobarbital and 5,6-benzoflavone pretreated liver S9 fractions from various animal species including rat, mouse, hamster and guinea pig. Stevioside and steviol at the concentrations up to 50 mg and 2 mg per plate, respectively showed no mutagenic effect on both tester strains either in the presence or absence of metabolic activating system. However, at the high concentration both stevioside and steviol showed some toxic effects on both tester strains. The toxic effect was decreased in the presence of the metabolic activating system. PMID- 9347661 TI - Effect of dialyzer membranes on beta-2 microglobulin production in Thai hemodialysis patients. AB - Responses to different types of dialyzer membranes in an Asian population may differ from those of a Caucasian population. Comparative studies on the effects of different dialyzer membranes on beta-2 microglobulin production are also limited. Therefore, we conducted this study to determine the effects of different dialyzer membranes on in vitro mononuclear cell production of beta-2 microglobulin in 9 Thai hemodialysis patients. Each patient was dialysed with 4 different types of dialyzer, including cuprophane (CUP), cellulose diacetate (CD), polysulphone (PS), and polyacrylonitrile membrane (PAN), each for a 1-month period in a randomized sequence. Mononuclear cell culture was done by taking an immediate post-dialysis blood sample at the end of the 1-month period. Beta-2 microglobulin production from cell culture was determined 24 hours later. Mononuclear cell culture and determination of beta-2 microglobulin production from the culture were also done in 10 normal controls and 10 predialysis ESRD patients. The beta-2 microglobulin productions (microgram/L) were shown as follows; Control CUP CD PS PAN [table: see text] (*p < 0.05 compared to cuprophane membrane). CONCLUSION: polysulphone and polyacrylonitrile membrane induced significantly less beta-2 microglobulin production compared to cuprophane and slightly less compared to cellulose diacetate membrane. PMID- 9347662 TI - Fever, skin rash, jaundice and lymphadenopathy after trichloroethylene exposure: a case report. AB - Trichloroethylene, a chlorinated hydrocarbon has been reported to cause many adverse health effects. This paper describes a female patient presenting with rather unusual manifestation secondary to trichloroethylene (TCE) exposure, i.e. hepatitis and generalized dermatitis. The diagnosis was confirmed by positive skin patch testing with 50 per cent TCE solution. After withdrawal from the exposure site, her symptoms improved and liver function test returned to baseline level after a three-months period of follow-up. TCE induced immunologic reaction has been postulated as the pathological process of this illness. PMID- 9347663 TI - Immunopharmacological activity of polysaccharide from the pericarb of mangosteen garcinia: phagocytic intracellular killing activities. AB - Polysaccharides from the pericarbs of mangosteen, Garcinia mangostana Linn., was obtained by treating the dried ground pericarbs with hot water followed by ethanol precipitation (M fraction). The extract was fractionated by anion exchange chromatography on a DEAE-cellulose column as MDE1-5 fractions. The fractions of MDE3 and MDE4 composed of mainly D-galacturonic acid and a small amount of neutral sugar (L-arabinose as the major one and L-rhamnose and D galactose as the minor ones) were studied for immunopharmacological activities by phagocytic test to intracellular bacteria (Salmonella enteritidis) and nitroblue tetrazolium (NBT) and superoxide generation tests. The results showed that the number of S. enteritidis in cultured monocyte with extract of pericarb of mangosteen (MDE3) was killed. Activating score (mean +/- SD) of NBT test of 100 polymorphonuclear phagocytic cells were 145 +/- 78, 338 +/- 58, 222 +/- 73, 209 +/- 77, 211 +/- 63, 372 +/- 19, 369 +/- 20, 355 +/- 34 in normal saline control, phorbol myristate acetate (PMA), MDE3, MDE4, indomethacin (I), PMA + MDE3, PMA + MDE4 and PMA + I, respectively. Superoxide generation test was also done by color reduction of cytochrome c. Both MDE3 and MDE4 stimulate superoxide production. The number of S. enteritidis in cultured monocyte with extract of pericarb of mangosteen was killed. This paper suggests that polysaccharides in the extract can stimulate phagocytic cells and kill intracellular bacteria (S. enteritidis). PMID- 9347665 TI - Association of mdr1 gene expression with other prognostic factors and clinical outcome in human breast cancer. AB - Multidrug resistance of cancer (CA) is one of a major problems in CA chemotherapy that is frequently associated with the expression of P-glycoprotein (P-gp) encoded by mdr1 genes. However, the controversial results exist regarding to the significance of mdr1 gene expression on clinical drug resistance to chemotherapy of breast CA cells. Recent evidence reported a strong correlation between the increased P-gp levels and the prognosis in advanced breast CA. The current study investigated whether mdr1 gene expression has any impact on prognosis and response to chemotherapy in breast CA patients. We determined mdr1 expression in 127 primary and 8 locally relapsed breast CA using a sensitive, specific and quantitative technique based on a RT-PCR and Southern blot hybridization detection by non-radioactive labelled-probe. In patients with primary breast CA, mdr1 expression were negative (mdr1-ve), low (< 10 units), high (> or = 10 units) in 63.8, 8.7 and 27.5 per cent of the patients, respectively. No differences in age, menopause status, tumor size, stage, lymph node involvement, estrogen receptor level and p53 level were observed between mdr1-ve and mdr1+ve expression patients. However, mdr1 gene expression is often associated with number of positive lymph nodes and negative estrogen receptors (p = 0.008 and 0.0007, respectively). In locally relapsed cases, mdr1-ve was 62.5 per cent whereas 37.5 per cent were mdr1+ve with high level of mdr1 RNA. No differences in other prognostic factors: lymph nodal involvement, estrogen receptor level and p53 level, were detected in both groups. Response to chemotherapy in primary and recurrent breast CA was not different in mdr1-ve and mdr1+ve patients. Finally, our results show that mdr1 gene expression is frequently present in breast CA both before and after chemotherapy. Association of mdr1 gene overexpression with other two prognostic factors suggests that they may confer a more aggressive nature of the tumor, drug resistance and poor prognosis. Evaluation of these factors may improve the ability to identify and select breast CA patients at high risk for poor prognosis for aggressive treatment. However, in this series response to CMF chemotherapy of primary and locally recurrent breast CA were not affected by the presence or absence of mdr1 gene product. PMID- 9347664 TI - Accuracy of fine needle aspiration cytology in the evaluation of peripheral lymphadenopathy. AB - We conducted an audit of the lymph node aspirates received from January 1996 to December 1996 of 541 patients sent to the Cytology Division, Department of Pathology, Ramathibodi Hospital by their clinicians. The aim of this retrospective study was to determine the pattern of diseases that commonly present with peripheral lymphadenopathy and to evaluate the accuracy of Fine Needle Aspiration Cytology (FNAC) in the diagnosis of lymph node diseases. An excisional biopsy sample of lymph node was available in 233 (43%) cases for comparison to the histopathology. The predominant lesion was benign which included necrotizing granulomatous lymphadenitis (NGL), reactive changes (RC) and suppurative lymphadenitis (SL). The predominant malignant lesion was metastatic squamous cell carcinoma. The accuracy for NGL, SL and RC were 69 per cent, 75 per cent and 95 per cent, respectively. The accuracy for metastatic disease was 97 per cent. The specificity and sensitivity of FNAC were 99 per cent and 94 per cent, respectively. An excisional biopsy should be done in case of doubt to clarify the pattern of RC. PMID- 9347666 TI - A study of epidemiology, risk factors and preventive measures against snake bites. AB - The epidemiology and exposing causes of snake bites were studied in 274 patients between 1 January 1982 and 30 December 1990. They comprised 142 males (51.82%) and 132 females (48.18%). Their ages ranged from 1 month to 86 years old. The age group most frequently bitten found in this study was between 15 to 29 years old amounting to 106 cases (38.69%). Of those 274 cases, 212 (77.37%) were the victims of poisonous snakes, namely, the green pit viper (Trimeresurus spp.) 156 cases (73.58%), the cobra (Naja naja) 53 cases (25.00%), Russell's viper (Vipera russellii siamensis) 3 cases (1.42%), while 13 cases (4.74%) were bitten by non poisonous snakes, and in the remaining 49 cases (17.88%), the biting snakes were not clearly seen. It was found that 56 cases (20.44%) were bitten in the house, 50 cases (18.25%) were bitten within the precincts of the residence. However the highest number, 168 cases (61.31%) were bitten outside. It was also interesting to learn that 221 cases (80.66%) out of 274 were bitten in Bangkok, 91 cases (33.21%) were bitten from 6.00 p.m. to 9.00 p.m., 129 cases (47.08%) were bitten between June and September, 98 cases (35.77%) were bitten on a foot and 77 cases (28.10%) were bitten on a finger. The high risk factor of being bitten by a snake was walking along a farm or a ranch with masses of grass or undergrowth during the night without self-prevention and good caution. Therefore, health education on preventive measures were elucidated. PMID- 9347667 TI - Sexual maturation in Thai girls. AB - Generally, menarche is considered an indicator of girls' sexual maturation. In Thailand, this study on the menarcheal age in female pupils was conducted in Bangkok, and the central, northern, northeastern and southern regions of Thailand, which differ in their geography, tradition and customs. In carrying out the research, 8,200 questionnaires were distributed to female pupils aged 10-17 years. The result was that the mean menarcheal age was 12.3 years. For the length of menstrual period, it was found that 83.7 per cent had a period of less than 7 days and 75.7 per cent used 2-4 pieces of sanitary napkins per day. For the regularity of menstrual period, 48.5 per cent had a regular period and 41.8 per cent had a period once within 2 months. For the persons whom these female pupils needed for advice about menarche, the study indicated that mothers were mostly needed. For the mean menarcheal age reported in each region, it was found that in the central region the mean menarcheal age was 12.5 years, the North was 12 years, the Northeast was 12.4 years and the South was 12.7 years. These data prove to be statistically significant, that is the lowest menarcheal age of female pupils was in the North while the highest was in the South. In terms of nature and characteristics of female pupils' menstruation in 4 regions there was no difference. When comparing the BMI value of female pupils already having a period in each region, the result was that there was no difference. The BMI value of those already having a period was higher than those not having had a period; and in the latter group, the BMI value of female pupils in the North was the highest while that in the Central region was higher than in the Northeast and the South. PMID- 9347668 TI - Press-fit-condylar total knee replacement: experience in 465 Thai patients. AB - A retrospective study of clinical results after primary total knee replacement using Press-Fit-Condylar (PFC) total knee arthroplasty was performed on 506 knees in 465 patients. There were 409 females and 56 males. Mean age at the time of surgery was 70 years (range; 31 to 86). The average follow-up time was 31 months (range; 6 to 67). The clinical results were graded according to the author's criteria which were based on functional requirements of the knee for the life style of Thai people. There were 115 excellent (painless stable knee with over 110 degrees of knee motion), 335 good (mild and occasional pain, medio-lateral instability less than 5 degrees and knee motion over 90 degrees), 46 fair (less severe persistent pain than pre-operative status, instability within 10 degrees and knee motion 60 degrees or more), 10 poor (pain not improved post-operatively, instability of over 10 degrees with range of motion under 60 degrees). Complications occurred in eight cases including: 3 infection, 3 patellar subluxation, one intra-operative popliteal artery injury and peri-prosthetic supracondylar fracture. No flexion contracture and loosening of the prostheses occurred in this study group. From this study, it was found that the PFC total knee replacement could provide satisfactory results of both short and intermediate clinical outcomes in terms of pain relief, stability and knee range of motion for Thai patients. PMID- 9347669 TI - Posterolateral lumbar fusion for degenerative spondylolisthesis: experiences of a modified technique without instrumentation. AB - From 1985 to 1994, 66 patients who had symptomatic degenerative spondylolisthesis of lumbar and lumbosacral levels managed by modified technique for decompressive laminectomy and postero-lateral spinal fusion. There were six males and sixty females with a mean age of 58.5 years (range, 47-73). Fifty- five patients had spondylolisthesis at L4-L5 level (51 were grade I and 4 were grade II), nine patients had spondylolisthesis at L5-S1 level (7 were grade I, 2 were grade II). There were two patients who had spondylolisthesis of both levels. Mean follow-up time was 48 months (range, 12-120 months). There were excellent and good results in 57 cases (86.4%), fair in 6 cases (9.1%). Three cases (5%) had poor results; 2 cases developed non-union, one case had L4 and L5 roots injury during operation. The overall fusion rate was 96 per cent. Solid spinal fusion occurred within 6 months. No further slip occurred in any cases in this study. PMID- 9347670 TI - Diagnostic hysteroscopy: a result of 125 patients at Ramathibodi Hospital. AB - This report summarizes the diagnostic hysteroscopic experience with 125 selected patients. The procedures were all performed under propofol anesthesia. The main indications for diagnostic hysteroscopy were infertility with suspected intrauterine lesions and abnormal uterine bleeding in premenopausal women. The procedures were successful in 123 (98.4%) patients. Cervical dilatation was required in 35 (28%) patients. Of the 125 diagnostic examinations, 91 (72.80%) had intrauterine abnormalities. This result showed that an important factor that appears to influence the prevalence of pathology are the gynecological problems and/or symptoms of the patients. The commonest finding in patients with infertility was intrauterine adhesions, whereas, endometrial polyps was the most common finding found in premenopausal women with abnormal uterine bleeding. There was no complication attributable to this procedure. Our experience suggests that the efficacy and safety of this procedure depend on proper selection of patients, type of anesthesia, the medium for uterine distention, and most importantly the experience of the operator. PMID- 9347671 TI - Percutaneous balloon pulmonary valvuloplasty in children: experience at Siriraj Hospital. AB - Twenty-seven children with pulmonary valvar stenosis with pressure gradient (PG) > or = 40 mmHg underwent percutaneous balloon pulmonary valvuloplasty (PBPV) at Siriraj Hospital between February 1993 and August 1996. There were 13 males and 14 females, with an age range from 2 months to 14 years, and body weight from 4.7 to 42.1 kg. The majority (92.6%) were asymptomatic. Before the PBPV, the pulmonary valve annulus (PVA) measured by echocardiography was significantly greater than that measured by cardiac catheterization (15.2 +/- 3.7 vs 14.5 +/- 3.9 mm, P = 0.006). However, there was linear association (r = 0.972) between the two methods. The PG obtained by the two methods showed no significant difference (90.8 +/- 35.3 by echocardiography vs 97.3 +/- 47.2 mmHg by catheterization, P = 0.266). Immediately after PBPV, the right ventricular systolic pressure (113.7 +/ 41.1 pre vs 62.3 +/- 28.1 mmHg post) and the PG 103.4 +/- 43.4 pre vs 49.0 +/- 31.1 mmHg post) were significantly reduced (p < or = 0.0005). At 6-mo follow-up echocardiography, the PG was 28.6 +/- 17.6 mmHg and was significantly reduced (P = 0.0005). The PVA significantly increased at the 12 mo follow-up (15.2 +/- 3.6 pre vs 17.6 +/- 3.8 mm post, P = 0.001). Only minor complications were reported in the present study; bleeding (3.7%), transient bradycardia (7.4%) and pulmonary regurgitation not more than moderate severity (79%). The immediate and intermediate results of PBPV are excellent with a success rate of 85 per cent. PMID- 9347672 TI - Amineptine in the treatment of amphetamine withdrawal: a placebo-controlled, randomised, double-blind study. AB - Temporary inability to function without amphetamine and the experience of withdrawal syndrome enhance the tendency for repetitive use. The investigators proposed to examine the therapeutic effects of amineptine, an antidepressant with dopamine reuptake inhibition effect, for the treatment of amphetamine withdrawal. The 14-day study was carried out on a randomised, double-blind, placebo controlled design. The authors assessed the severity of amphetamine withdrawal syndrome by using two measures and performed both end-point and intent-to-treat analyses. The results showed that amineptine helped relieve a depressed mood within one week and improved the general condition within 2 weeks. In conclusion, amineptine is effective in rapid relief of depressed mood and improves the general condition of patients with amphetamine withdrawal. Since the amphetamine withdrawal may last for several weeks, studies with longer duration should be conducted before incorporating amineptine into the clinical practice a of amphetamine withdrawal treatment. PMID- 9347673 TI - Topical analgesics for knee arthrosis: a parallel study of ketoprofen gel and diclofenac emulgel. AB - The clinical efficacy of the two topical analgesics, ketoprofen hydroalcoholic gel (Fastum gel) and diclofenac emulgel, for osteoarthritis of the knee was studied. There were 85 patients who underwent the trial. They were randomly allocated into 2 groups, the diclofenac group, 42 patients (4 males and 38 females) receiving the diclofenac emulgel at the painfull site four times a day for 4 weeks, and the ketoprofen group, 43 patients (9 males and 34 females) receiving the ketoprofen hydroalcoholic gel four times a day for 4 weeks at the painful knee. Golberg's knee scoring was used to evaluate the patients before the trial, at the end of the first, second and fourth weeks. The ketoprofen group had poorer a score before the trial, however, both groups had improvement in their knee functions, knee score and pain. There was no significant difference between the groups at the end of the study. There was no serious side effect in both groups. Ketoprofen hydroalcoholic gel gave persuasive results in the treatment of knee arthrosis stage I and II. PMID- 9347674 TI - Differentiation of Anopheles minimus species complex by RAPD-PCR technique. AB - Amplification of random regions of genomic DNA using 10-base primers in the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to differentiate Anopheles minimus A and An. minimus C. Genomic DNA was extracted from individual mosquitoes of An. minimus A and An. minimus C and amplified in PCR reactions using single primers of arbitrary nucleotide sequence. Fifteen different commercially available primers (Operon oligonucleotides kit M from Operon Technologies, Inc.), six primers were selected on the basis of presence or absence of the bands of A and C. They gave 8 different amplified DNA fragments of these two species of An. minimus. The primers revealed only species A are OPM8 and OPM13 at the 0.8 and 2.15 and 2, while both species A and C were in OPM17 at 0.8 Kb revealed A, at the 0.55 and 1.5 Kb revealed C. OPM 12 gave a 0.5 Kb DNA fragment in A while 0.4 Kb in C. OPM15 showed C at 0.7 Kb. These findings indicated that An. minimus species A and C can be differentiated by RAPD-PCR technique. PMID- 9347675 TI - Cerebellopontine angle meningiomas with primary otologic symptoms. AB - The majority of cerebellopontine angle (CPA) tumors are acoustic neuromas (AN). However, an intracranial meningioma may occur at this site and will produce symptoms similar to an AN. The most common presenting symptoms of CPA meningiomas are hearing loss, tinnitus, dizziness and dysequilibrium. It cannot be easily distinquished from an AN only on the history and physical examination. Even with an audiogram, evoked response audiometry (ERA) and vestibular function tests, it still cannot be distinquished. CT scan and MRI are helpful in differentiating these two tumors radiographically. In this article, we report two cases of CPA meningiomas which presented with otologic symptoms. The diagnosis and treatment of CPA meningioma is discussed. PMID- 9347676 TI - Management of celiac artery aneurysm: a case report. AB - A case of CAA in a 68-year-old male patient is reported. The patient had vague abdominal pain for 1 year. Physical examination revealed a pulsatile abdominal mass on the epigastrium. Abdominal ultrasonography and visceral arteriography confirmed the diagnosis of CAA. Aneurysmectomy with direct implantation was performed transabdominally. PMID- 9347677 TI - Low-dose oral interferon-alpha: effective prophylaxis for gingivitis and aphthous ulcers in AIDS patients. PMID- 9347678 TI - HIV and the black community: the role of the National Medical Association. PMID- 9347679 TI - Community violence: causes, prevention, and intervention. AB - This article presents some pragmatic schemata for understanding various types and motivations for violence. This understanding is essential to frame prevention, intervention, and postvention strategies designed to reduce the phenomena of violence in our society. Each category of violence lists examples of prevention, intervention, and postvention strategies. This article is intended to broaden the understanding of violence so that strategies to address violence will become more specific and measurable. PMID- 9347680 TI - Causes of chest pain and symptoms suggestive of acute cardiac ischemia in African American patients presenting to the emergency department: a multicenter study. AB - This study examines whether race is a significant determinant of the diagnoses of acute myocardial infarction or angina pectoris in patients with symptoms suggestive of acute cardiac ischemia. The study population was comprised of 3401 (34%) African-American and 6600 (66%) white patients who presented to emergency departments with symptoms suggestive of acute cardiac ischemia. The main outcome measure was a diagnosis of acute myocardial infarction or angina pectoris. African Americans were younger, predominantly female, and more often had hypertension, diabetes mellitus, or smoked. The diagnosis of acute myocardial infarction was confirmed in 6% of African-American and 12% of white men, and in 4% of African-American and 8% of white women. After adjusting for age, gender, medical history, signs and symptoms, and hospital, African Americans were half as likely to develop acute myocardial infarction and were 60% as likely to have acute cardiac ischemia. Despite having less acute cardiac ischemia, African Americans in this study had high risk levels for coronary artery disease. PMID- 9347682 TI - Ophthalmological morbidity in very-low-birthweight infants with bronchopulmonary dysplasia. AB - This study was undertaken to determine the relationship between retinopathy of prematurity, ocular sequelae of retinopathy, and bronchopulmonary dysplasia in infants weighing < 1250 g at birth prior to the introduction of steroid therapy for chronic lung disease. Ophthalmological data from 67 infants (22 with severe bronchopulmonary dysplasia and 45 controls) who were enrolled prospectively in an early intervention program were analyzed. The infants had two or more eye examinations prior to discharge and a follow-up examination at 12 to 18 months postconceptual age. The incidence of any retinopathy of prematurity was 33%, and severe retinopathy was 25%. Infants with severe bronchopulmonary dysplasia were 1.7 times more likely to develop any retinopathy and 1.8 times more likely to develop severe retinopathy than controls. The incidence of ocular sequelae, was 45%. Infants with any retinopathy had a 2.3 odds of developing sequelae, and infants with severe retinopathy had a 2.64 odds ratio. When adjusted for bronchopulmonary dysplasia, the odds ratio for developing sequelae was 1.36 in infants with any retinopathy and 1.27 in those with severe retinopathy. The predictors of retinopathy were lower birthweight and gestational age, acidosis, and hypoxemia. Bronchopulmonary dysplasia per se has an adverse effect on ophthalmologic morbidity. Evaluation of the adverse effect of any therapy for chronic lung disease on retinopathy of prematurity should make adjustments for the underlying lung disease. PMID- 9347681 TI - Modulation of lipid profile by fish oil and garlic combination. AB - Fish consumption has been shown to influence epidemiology of heart disease, and garlic has been shown to influence triglyceride levels. This study was undertaken to evaluate the effect of fish oil and garlic combinations as a dietary supplement on the lipid subfractions. Forty consecutive subjects with lipid profile abnormalities were enrolled in a single-blind, placebo-controlled crossover study. Each subject received placebo for 1 month and fish oil (1800 mg of eicosapentanoic acid [EPA] + 1200 mg of docosahexanoic acid) with garlic powder (1200 mg) capsules daily for 1 month. Lipid fractionation was performed prior to study initiation, after the placebo period, and after the intervention period. Subjects all had cholesterol levels > 200. Subjects were instructed to maintain their usual diets. Supplementation for 1 month resulted in an 11% decrease in cholesterol, a 34% decrease in triglyceride, and a 10% decrease in low-density lipoprotein (LDL) levels, as well as a 19% decrease in cholesterol/high-density lipoprotein (HDL) risk. Although not significant, there was a trend toward increase in HDL. There was no significant placebo effect. These results suggest that in addition to the known anticoagulant and antioxidant properties of both fish oil and garlic, the combination causes favorable shifts in the lipid subfractions within 1 month. Triglycerides are affected to the largest extent. The cholesterol lowering and improvement in lipid/HDL risk ratios suggests that these combinations may have antiatherosclerotic properties and may protect against the development of coronary artery disease. PMID- 9347684 TI - Organ/tissue donation the problem! Education the solution: a review. PMID- 9347683 TI - Caregiver issues and AIDS orphans: perspectives from a social worker focus group. AB - This study examines social workers' perceptions of the needs of families coping with acquired immunodeficiency syndrome (AIDS). This research investigates the problems of family caregivers of children orphaned by human immunodeficiency virus (HIV)-related death of their parents. A qualitative semistructured interview format was used in a focus group of 18 social workers. Four questions were designed to assess family needs and resources, as well as to evaluate the social workers' perspectives of governmental policies affecting these families. A list of four problems and two recommendations for change evolved from the focus group. Inadequate finances to house and care for the children was the primary cause for distress in these families. The major governmental policy that hindered the social workers' ability to assist families pertained to the low financial entitlement for caregivers who are related to the orphaned child. It was noted that unrelated caregivers receive substantially more money for the care of these children than family caregivers receive. Recommendations were made to change this policy and to develop guardianship laws that facilitate families' abilities to provide care to AIDS orphans. Family caregivers of AIDS orphans are bombarded with great demands and limited resources. This analysis of their situation from the social workers' perspective is a positive step toward the improvement of support services for these families. Further research should include individual qualitative interviews assessing the needs of the caregivers and AIDS orphans. PMID- 9347686 TI - Africa and the academic pipeline: cultural reflections and celebration as motivational tools for black school children. PMID- 9347685 TI - Use of fluconazole in the treatment of non-AIDS cryptococcal meningitis. AB - The treatment of non-acquired immunodeficiency syndrome (AIDS) cryptococcal meningitis has not been well defined as yet. The current recommendation is amphotericin B with or without flucytosine. However, due to the many side effects of these drugs, fluconazole is becoming an attractive alternative in patients who cannot tolerate amphotericin B and flucytosine. This case study demonstrates the successful use of fluconazole in a patient with underlying diabetes mellitus and cryptococcal meningitis. PMID- 9347687 TI - Comparative characteristics of some cultural and toxigenic properties of the Dendrodochium Bonorden and Myrothecium Tode ex Fr. representatives. AB - The comparative study of sporulation intensity, change of medium pH, yield of biomass and ethanol soluble fraction (ESF) of extracellular metabolites as well as antibiotic and toxigenic properties of 106 fungi strains which were the potential producers of macrocyclic trichothecenes has been carried out under the conditions of stationary cultivation. It was shown that all the studied Dendrodochium strains were characterized by moderate growth, variations of biomass were within 3.5-4.5 g/l. Variations of ESF level in different strains were rather essential (from 8 to 241.6 mg/l). High antibiotic activity correlated with high or moderate toxicity (on the basis of rabbit skin test) in most cases. Dendrodochium strains with intensive sporulation were as a rule characterized by high efficiency of toxin production (40% of studied strains) and only 3 strains (5.5%) with low level of sporulation were characterized by high rate of toxin production. The toxigenic activity for Myrothecium cinctum, M. commune and M. ukrainicum has been shown for the first time. The toxigenic activity of the studied strains depending on the source of isolation and possible correlative connections between sporulation, toxin formation and other properties are discussed. PMID- 9347688 TI - Certain peculiarities of biological action of the stachybotryotoxin preparations. AB - Some biological and toxigenic properties of 47 Stachybotrys alternans strains, isolated from different ecological niches have been studied. On the basis of antifungal, phytotoxic and dermacidic properties the strains studied can be divided into several characteristic groups: strains with clearly expressed dermacidic reaction and without other properties studied (19.1%); strains displayed only antifungal activity to a number of yeast test cultures (4.3%); strains with complex action (antifungal, phytotoxic and dermacidic activity) (46.8%). Considerable portion of strains (25.5%) did not show any biocidal activity tested. No clear dependence of strain activity on the term of storage under laboratory conditions (from 5 to 40 years) has been revealed. PMID- 9347690 TI - Life in the ER: on the front line of medicine. AB - Emergency medicine is the new kid on the block. It was not officially recognized as a medical specialty until 1979, and among all medical specialties, it is the second youngest. The first residency program in emergency medicine began in 1970 at the University of Cincinnati--with only one resident. PMID- 9347689 TI - Specific identification of Mycobacterium bovis by monoclonal antibody-based enzyme immunoassay. AB - To improve identification of M. bovis, rabbit immune sera and mouse monoclonal antibodies against M. bovis-secreted protein antigens were used in the enzyme linked assay (ELISA). In western blot analysis, M. bovis-specific epitopes within culture filtrate proteins were demonstrated to be mostly concentrated on the immunodominant 35-38 kDa antigen. Polyclonal and cross-reactive monoclonal antibodies were shown to be able to bind to all mycobacterial strains tested in ELISA (M. tuberculosis, M. bovis, M. kansasii, M. marinum, M. avium, M. smegmatis), but not to bacteria of the other genera. Among the monoclonal antibodies against M. bovis specific antigen, 2-6B was found to be the only one which could evidently react with whole cells of M. bovis in ELISA, but not with those of the other mycobacteria including M. tuberculosis. PMID- 9347691 TI - The race for governor: talking about health care. AB - During the 1989 gubernatorial race, Jim Florio liked to say that a political campaign is a conversation--a time for candidates and voters to communicate aspirations and concerns, an opportunity for political engagement and dialogue. An election campaign is a time when citizens pay more attention to politics, and candidates have more personal contact with voters. For both citizens and candidates, a campaign at its best is a time for education. PMID- 9347692 TI - Providing influenza and pneumococcal disease vaccinations. PMID- 9347693 TI - Radiology/pathology conference at UMDNJ. PMID- 9347694 TI - Domestic violence: break the silence. PMID- 9347695 TI - From hospital to home: discharge planning nurses. PMID- 9347697 TI - Telemedicine. Slow in coming. PMID- 9347696 TI - Supreme court challenges New Jersey physicians in suicide cases. PMID- 9347698 TI - Practicing medicine: liability lessons. PMID- 9347699 TI - Response to epidemic meningitis in Africa, 1997. PMID- 9347701 TI - Model still needed for defining and evaluating quality of ambulatory care education. PMID- 9347700 TI - Physicians must confront ageism. PMID- 9347702 TI - Physicians need more education about inherited cancer predisposition. PMID- 9347704 TI - Academic medicine and the issues of aging. PMID- 9347703 TI - Why Argentinian students studied medicine then and now. PMID- 9347705 TI - Pressure from our aging population will broaden our understanding of medicine. AB - This essay speculates about how medical philosophy may change to accommodate the technologic, organizational, and--in particular--population challenges of the future. The historical roots of the dominant medical world view are drawn and its tenets outlined. The existing paradigm may be called the Western biomedical model, whose doctrines include body-mind dualism, physical reductionism, the mechanical analogy, specific etiology, and the body as the appropriate object of regimen and control. Some of the pressures straining the paradigm are discussed, especially the force of human and population aging as it accelerates the dominance of chronic illness and focus of health care. The tentative outlines of an emergent paradigm are described. The mind, biography, surrounding environment, and culture are a few considerations that become very significant in a non Cartesian world. PMID- 9347706 TI - Reflections on death and dying. AB - Americans simultaneously worry about dying and about being tethered to machines that keep them alive beyond a point when life has any meaning. People living with terminal illness often feel isolated from life around them and a burden on those they love; they feel uncertain that their deaths will be relatively free of pain and suffering and that their dignity will be compromised as little as possible. These failings can be remedied. Traditional hospice care and integrating palliative care into the general medical setting are important, but they cannot alone occasion a better dying. The medical community must re-imagine dying and reflect about ways to transform image into reality in practice and in training colleagues and successors. Physicians and others know how to provide care and even improve living when cure is unlikely; the harder task is to respect such care as profoundly as curing. The exigencies of modern medicine, where time is a budgetable commodity, makes caring well for dying patients difficult. Medicine cannot have hegemony over dying and cannot singularly offer people a better death, but it cannot absent itself either. The almost single-minded focus on decision making that has infused conversations about dying and death may divert attention from the attentiveness and loving relationships that are as vital as life's end as at its beginning. Medicine has "colonized" death: It has transformed it into a place where progress in staving it off may appear to be unlimited, and thus it encourages forgetting that death is part of the human condition. The task before medicine, and academic medicine in particular, is to transform death back into a human scale. With all that is available to delay death--but not to make it optional--the most important task is to recover humbleness before an awesome moment and be with the patient, one human being to another, knowing that dying is not always open to solutions. PMID- 9347707 TI - Questions for societies with "Third Age" populations. The Extension-of-Life Working Group, The Gerontological Society of America. AB - By 2030, the number of people aged 65 and over in the United States will total 70.2 million, 20% of the U.S. population, making the older population a major consumer of goods and services. The demands of this growing group are already affecting the medical professional, changing the mix of patients, conditions treated, and what the profession and society consider acceptable outcomes. Medical education will have to adjust to these new demands by training in the diagnosis and treatment of older persons, by equating successful management of chronic conditions with curing illnesses or fixing hurts, and by dealing with the inevitability of death. Also, however, medical education should train students to understand that the rapid aging of the population has the potential to affect almost every human and societal arrangement and every social and economic institution, raising a host of ethical, moral, scientific, social, and economic questions. The Gerontological Society of America (GSA) has convened an informal working group to explore the issues raised by the extension of human life expectancy. The multidisciplinary group, which has 13 members from academia and the GSA, held its first meeting in May 1997, when it laid out the issues to be discussed in future meetings. To help frame the discussions, the group adopted the concept of the "Third Age," a time in the life course when for most people the basic work of parenting is done, when, under current arrangements and definitions, people are not heavily relied on for production (that is, a time when people for different reasons leave paid, full-time jobs), and when few positive roles are recognized. The group's discussions will explore sets of interrelated questions raised by longer life expectancy and the larger number of older people. These questions can be considered in five general themes: the further extension of human life expectancy, research choices, societal vision and values, global aging, and economies and Third Age populations. These questions are raised in the paper, and in coming months the group will discuss their implications. PMID- 9347708 TI - Aging research and education centers in the United States: a compendium. AB - U.S. centers and institutes for research and education devoted to aging are listed. These lists can serve as a starting point for building a more comprehensive reference resource. The first list, U.S. Aging Centers and Institutes, is a general guide to centers or institutes that combine research and education. Subsequent lists are of centers that share missions and funding sources: Geriatric Research, Education and Clinical Centers (GRECCs); Exploratory Centers for Research on Health Promotion in Older Minority Populations; Centers on the Demography of Aging (CDAs); Alzheimer's Disease Centers (ADCs); Claude D. Pepper Older Americans Independence Centers (OAICs); Nathan Shock Centers of Excellence in Basic Biology of Aging; and Roybal Centers for Research on Applied Gerontology. It is hoped that those who work in geriatrics and gerontology in academic medicine will develop a comprehensive system for collecting, updating, and disseminating complete information about the work being done on aging. PMID- 9347709 TI - Cutting the Gordian knot: a two-part approach to the evaluation and professional development of residents. AB - There is chronic dissatisfaction among both faculty and students about the process and effectiveness of resident performance evaluation. The author asserts that the source of this problem is the current practice of merging the two different purposes for evaluation: to monitor residents' meeting of performance standards and to provide guidance for residents' professional development. By attempting to meet both quality-control and guidance obligations using one set of objective data, most residency programs fall short in meeting one of these aims. The common preoccupation with psychometric precision, objectivity, and the statistical processing of forms frequently distracts users from making effective use of evaluation information. The proposed solution is to divide resident evaluation into two simpler, entirely separate and distinct systems--neither of which would look much like the current system. There would be a faculty controlled, quality-control system focused on screening for minimal performance standards. This would use simple, qualitative measures for early warning and rapid follow-up. The second evaluation system would be a resident-controlled, guidance-oriented system focused on self-assessment, peer and faculty coaching, and reflection. The hypothesized benefits of this approach include an improvement in residents' motivation and performance, an increase in residents' self direction, and an enhancement of communication between residents and faculty members. PMID- 9347710 TI - A human-centered approach to medical informatics for medical students, residents, and practicing clinicians. AB - The authors have developed a curriculum in medical informatics that focuses on practical problems in clinical medicine, rather than on the details of informatics technologies. Their development of this human-centered curriculum was guided by the identification of six key clinical challenges that must be addressed by practitioners in the near future and by an examination of the failures of past informatics efforts to make a significant difference in the everyday practice of clinical medicine. Principles of human factors engineering- the body of knowledge about those human abilities, limitations, and characteristics that are relevant to design--are an essential part of this curriculum. Human factors engineering also provides the necessary perspective, as well as the concrete knowledge and methods, that can enable practitioners to properly evaluate their clinical information needs, weight the merits of proposed technology-based solutions, and understand their own inherent performance limitations. PMID- 9347711 TI - Meeting the challenges of an aging population. PMID- 9347712 TI - Downsizing of basic science departments in U. S. medical schools: perceptions of their chairs. The National Caucus of Basic Biomedical Science Chairs. AB - PURPOSE: To assess the status of basic science departments in academic health centers through a survey of department chairs. METHOD: In November 1995 the National Caucus of Basic Biomedical Science Chairs developed a 35-item questionnaire that was sent to the presidents of the Caucus's six member associations, who distributed it to their medical school chairs. The questionnaire was sent to 90% of the basic science chairs; a total of 683 questionnaires were distributed. By April 1996, 59% (400/683) of the chairs responded. Preliminary tabulations were made by each association, and then all the questionnaires were sent to the Caucus chair for compilation and analysis. RESULTS: The chairs in all six basic science disciplines reported four basic trends. Reduction in staffing: Eleven percent of the chairs were in "acting" positions, temporarily filling vacancies that sometimes had not been filled for five years or more. There were 583 reported faculty vacancies, but active searches were under way for only 339 of these. The chairs reported the abolition of, on average, one faculty position per department between 1985 and 1995, but these losses were clustered in only a third of the schools; thus several departments had experienced extensive faculty depletion. Overall, there was a 10% attrition of basic science faculty. Restructuring of the basic sciences: In all, 22 disciplinary departments had merged between 1985 and 1995, and merger discussions were under way at another 9% of the departments. Seven percent of the chairs reported that their schools were considering a single, combined basic science department, but only one such unit exists so far. Graduate student recruitment had become solely interdisciplinary at 26% of the schools, had remained strictly departmental at another 44%, and was joint at the remaining 30%. The development of a non-research-based teaching track was being considered at a fourth of the reporting departments. New fiscal policies/professional stability: A salary incentive beyond the university contribution, to reward research funding, was in effect at 28% of the departments and under consideration by another 29%. For a junior faculty member who, at the time of tenure decision, lacked research grant support, termination was reported by almost a third of the chairs as being extremely likely; however, half of the chairs did not respond to this question. Changes in tenure: A fourth of the chairs reported that modifications of their school's tenure policies were under serious consideration; and almost three fourths felt that tenure had become more difficult to achieve. CONCLUSION: This survey reveals certain profound and disconcerting changes in the basic science departments of U.S. medical school. More studies are needed to see whether these trends continue. Further downsizing and merging of the basic sciences should be avoided in order to preserve the quality of medical education for future physicians as well as the quality of health care in the United States. PMID- 9347713 TI - Academic deans' views on curriculum content in medical schools. AB - PURPOSE: To determine which of 33 topics academic deans identify as worthy of greater emphasis in medical curricula. Also, to assess the barriers to needed curricular changes. METHOD: In March 1996 a questionnaire was developed and mailed to the academic deans of all U.S. schools affiliated with the Association of American Medical Colleges (n = 126) and all schools associated with the American Association of Colleges of Osteopathy (n = 17). There were 46 questions in a five-point Likert-type format (1 = not at all, 5 = to a great extent) and one open-ended question. The deans were queried as to what extent each of 33 topics (1) was included in medical students' required learning experiences (current emphasis) and (2) should be included in medical students' required learning experiences (ideal emphasis). The deans were also asked to what extent they believed 12 different factors would be barriers to needed curriculum changes in their programs. Primary data analysis focused on simple comparisons of response means and frequencies. RESULTS: Two separate mailings resulted in the return of 100 questionnaires (70%): 85 from the allopathic schools (67%) and 15 from the osteopathic schools (88%). "Effective patient-provider relationships/communication," "outpatient/ambulatory care," and "health promotion/disease prevention" had the three highest mean ratings for ideal emphasis by the allopathic school deans. "Primary care," "professional values," and "use of electronic information systems" also had high mean rankings for ideal emphasis. "Primary care," "outpatient/ambulatory care," and "health promotion/disease prevention" had the three highest mean ratings for ideal emphasis by the osteopathic school deans. CONCLUSION: Changes in health care delivery and an increasing generalist orientation are influencing academic deans' perspectives on needed curriculum changes, and there appears to be considerable support for medical school curricula that will foster a broader, more humanistic role for physicians. PMID- 9347714 TI - Predicting who will choose a family medicine residency. AB - PURPOSE: To create and evaluate a screening instrument and a revised interview format for use in the selection of family medicine residents at the McGill University Faculty of Medicine. METHOD: The screening tool consisted of two sections an assessment of academic performance (the TASS) and an evaluation of applicants' generalist versus specialist orientation (the GSSS); each applicant's file was assessed by two raters. The revised interview included specific questions and scenarios related to family medicine. All three parts were tested on 143 applicants from outside the region of Quebec in 1994-95. The results on both parts of the screening tool and the interview were compared with the students' first choices of residency and then were compared with the performances of the 24 accepted into the program as reflected in their first six-month summative evaluation forms. Data were analyzed through several statistical methods. RESULTS: Only the GSSS accurately predicted the applicants' first choices (for agreement between both raters: sensitivity, 81%; specificity, 70%; accuracy, 78%). No significant association was found when comparing matching applicants' scores obtained during the selection process with their scores on the six-month evaluation forms. CONCLUSION: The GSSS may prove useful as a tool in the review of applicants' files. More studies are needed to reevaluate the use of the interview in the selection process and to help determine whether any selection instrument can accurately predict applicants' subsequent performances in a residency. PMID- 9347715 TI - The effect of incorporating women's health into a PBL curriculum on students' tendencies to identify learning issues in an ambulatory care setting. AB - PURPOSE: To investigate whether the incorporation of women's health into problem based learning (PBL) cases affects students' tendency to identify learning issues related to women's health as they encounter patients in an ambulatory care setting. METHOD: Students in the PBL curriculum at the Allegheny University of the Health Sciences, MCP-Hahnemann School of Medicine, participate in a nine-week primary care practicum at the end of their first year, during which they spend three half-days per week in an ambulatory setting examining patients and completing patient logs that include any learning issues identified. Patient logs from 23 first-year PBL students who had not been exposed to a new women's health education program prior to their practicum in 1993 and from 22 first-year PBL students who had been exposed to the program prior to their practicum in 1994 were reviewed. For each women's health learning issue identified, the sex of the student and the sex, specialty, and practice setting of the student's preceptor were recorded. Data were analyzed with several statistical methods. RESULTS: There was no statistically significant difference in the numbers of men and women students or preceptors between the two years. In 1993 an average of 59% of the patients seen per student were women; in 1994 the average was 61%. The mean numbers of total learning issues identified (including women's health learning issues) were similar in the two years, but the mean percentage of clinical women's health learning issues identified increased significantly between 1993 and 1994, as did the mean percentage of community/preventive health women's health learning issues identified. There was a significant student-sex-by preceptor-sex interaction for the total number of women's health learning issues identified (p = .024): for both years, the students paired with a preceptor of the same sex identified a higher number of women's health learning issues than did the students paired with a preceptor of the opposite sex. CONCLUSION: The results suggest that PBL is an effective way to increase students' awareness of women's health issues in a primary care clinical setting. More studies are needed to define the effect of PBL on the kind of reading and learning students will do when they get to the clinical setting. PMID- 9347717 TI - A pilot study to determine the effect of patient educators on medical students' and residents' skills in joint examination. PMID- 9347718 TI - The relationship of students' admission interview ratings to their performances in a medical-interviewing course. PMID- 9347716 TI - The effect of educational gifts from pharmaceutical firms on medical students' recall of company names or products. AB - PURPOSE: To assess the influence of pharmaceutical advertising (in the form of books) directed at medical students and also to examine students' attitudes toward pharmaceutical representatives after interacting with them. METHOD: Two groups of fourth-year medical students were surveyed: 166 residency applicants to the Department of Anesthesia and Critical Care between 1991 and 1993, who were questioned during their personal interviews with the department chair, and 39 fourth-year students from the University of Chicago Pritzker School of Medicine in 1994-95, who were surveyed by telephone. The students were asked if they had ever received a book from a pharmaceutical representative and, if so, to name the book. Then they were asked to name the book-giving company or a product associated with the company. Responses were compared using chi-square analysis. RESULTS: In all, 90% of the students had received one or more books and accurately recalled titles for 89% of them. However, only 25% of the named books were accurately associated with a pharmaceutical company or product. The Pritzker students, asked to recall interactions with pharmaceutical representatives, reported being skeptical of representatives who ignored them because they were students, but they rated as helpful and informative those who conversed with them or gave them gifts. CONCLUSION: Although gifts to medical students do not necessarily engender company or product recall, attention paid to medical students by pharmaceutical representatives engenders goodwill toward the representatives and their messages. PMID- 9347719 TI - Baseline results on the ATSIM before the introduction of a revised medical curriculum. PMID- 9347720 TI - Patients transferred to academic medical centers and other hospitals: characteristics, resource use, and outcomes. AB - As purchasers of health care increasingly rely on hospital resource use and outcomes profiles to guide quality improvement efforts and contract decisions, a better understanding of the contribution of the most severely ill patients to aggregate resource use and outcomes is needed to develop measures that make fair comparisons between hospitals. In this article, the authors examine the distribution, resource utilization, and outcomes of transferred patients ("transfers"), a group known to be highly complex. The study examines the contributions to resource use and outcomes of these patients at academic medical centers (AMCs) and non-teaching hospitals. The authors go beyond previous work by comparing AMCs with non-teaching hospitals, and by using a nationally representative sample for the year 1992. The detailed findings demonstrated that AMCs provided a disproportionate share of care to transfers in 1992, and that transfers to AMCs are more complex and require more specialized care than do transfers to non-teaching hospitals. The study also determined that, during the time studied, AMCs received a disproportionate share of Medicaid and indigentcare transfers. Finally, the findings demonstrated that transfers increased in absolute numbers and as a percentage of total patient volumes for all hospitals from 1988 to 1994. The rate of increase was greatest for AMCs. The authors explain why they believe that their findings are applicable today, although they caution that study of more recent data should be made. The authors comment that purchasers of health care may find the study useful in better understanding benchmarking tools used to evaluate hospitals. This study may also help those involved in health policy to more fully understand the magnitude of the contribution to transfer patient care provided by AMCs. Finally, health policymakers and planners may find this work useful as they prepare for increasing numbers of transfers in the future, particularly at AMCs. PMID- 9347721 TI - Attitudes toward physician-nurse alliance: comparisons of medical and nursing students. PMID- 9347722 TI - Labeling each response option and the direction of the positive options impacts student course ratings. PMID- 9347723 TI - Prioritizing areas for faculty development of clinical teachers by using student evaluations for evidence-based decisions. PMID- 9347725 TI - Medical student perception of the academic environment: a prospective comparison of traditional and problem-based curricula. PMID- 9347724 TI - The effect of PBL on students' perceptions of learning environment. PMID- 9347726 TI - Can non-expert PBL tutors predict their students' achievement? An exploratory study. PMID- 9347727 TI - Item dependency in medical licensing examinations. PMID- 9347728 TI - Tests of sequential testing in two years' results of Part 2 of the Medical Council of Canada Qualifying Examination. PMID- 9347729 TI - Sequential testing in the objective structured clinical examination: selecting items for the screen. PMID- 9347730 TI - Longitudinal effectiveness of a medical school geriatrics clerkship. PMID- 9347731 TI - Predicting performances on the NBME Surgery Subject Test and USMLE Step 2: the effects of surgery clerkship timing and length. PMID- 9347732 TI - The effects of psychiatry clerkship timing and length on measures of performance. PMID- 9347733 TI - Assessing the semantic content of clinical case presentations: studies of reliability and concurrent validity. PMID- 9347735 TI - Use of encounter cards for evaluation of residents in obstetrics. PMID- 9347734 TI - The effect of presentation order in clinical decision making. PMID- 9347736 TI - Improving the quality of resident performance appraisals. PMID- 9347737 TI - Using an OSCE to assess the ability of residents to manage problems in women's health. PMID- 9347738 TI - Do residents also feel "abused"? Perceived mistreatment during internship. PMID- 9347739 TI - "Turfing" narratives and the ideology of residency. PMID- 9347740 TI - Medical students' self-assessment accuracy in communication skills. PMID- 9347741 TI - An interdisciplinary approach to teaching clinical laboratory skills to medical students. PMID- 9347742 TI - Accuracy of second-year medical students' self-assessment of clinical skills. PMID- 9347743 TI - Academic performances of early-admission students to a BA/MD program compared with regular-admission students in relation to applicant pool fluctuations. PMID- 9347744 TI - Cognitive and noncognitive predictors of academic difficulty and attrition. PMID- 9347745 TI - An analysis of admission committee voting patterns. PMID- 9347746 TI - An evaluation of the Rasch model for equating multiple forms of a performance assessment of physicians' patient-management skills. PMID- 9347747 TI - Issues of validity and reliability concerning who scores the post-encounter patient-progress note. PMID- 9347748 TI - Feasibility and psychometric properties of using peers, consulting physicians, co workers, and patients to assess physicians. PMID- 9347749 TI - Standardized patients' accuracy in recording examinees' behaviors using checklists. PMID- 9347750 TI - Assessing the factor structure of a nationally administered standardized patient examination. PMID- 9347752 TI - The usefulness of test-performance feedback in preparing to repeat the USMLE Step 3 Examination. PMID- 9347751 TI - Using standardized patients to ensure that clinical learning objectives for the breast examination are met. PMID- 9347753 TI - Medical students' reasons for changing answers on multiple-choice tests. PMID- 9347754 TI - A comparison of retest performances and test-preparation methods for MCAT examinees grouped by gender and race-ethnicity. PMID- 9347755 TI - Generalist career plants: tracking medical school seniors through residency. PMID- 9347756 TI - Primary care residency selection and completion--the Illinois experience, 1988 1995. PMID- 9347757 TI - Factors influencing primary care physicians' choice to practice in medically underserved areas. PMID- 9347758 TI - Profile of medicine clerkship directors. PMID- 9347759 TI - How much service, education, and research? An empirical study in a university teaching hospital. PMID- 9347760 TI - Relationships of how well attending physicians teach to their students' performances and residency choices. PMID- 9347761 TI - Small-group teaching of chest auscultation to third-year medical students. PMID- 9347762 TI - Computer-assisted instruction for the medical histology course at SUNY at Buffalo. PMID- 9347763 TI - Comparing how well two undergraduate curricula teach knowledge of internal medicine. PMID- 9347764 TI - Comparison of ambulatory knowledge of third-year students who learned in ambulatory settings with that of students who learned in inpatient settings. PMID- 9347765 TI - Ambulatory care education in an emergency room setting: effects of house officer specialty and prior experience. PMID- 9347766 TI - Educational correlates of students' perceptions of learning in longitudinal ambulatory primary care clerkships. PMID- 9347767 TI - A thematic review of the literature of underrepresented minorities and medical training, 1981-1995: securing the foundations of the bridge to diversity. PMID- 9347768 TI - Medicine and the arts: on aging. PMID- 9347769 TI - Migraine research methods. PMID- 9347770 TI - Ruling out spinal fractures in trauma. PMID- 9347771 TI - Terfenadine and SSRIs. PMID- 9347772 TI - The placebo effect. PMID- 9347773 TI - The health of Canada's elderly population: current status and future implications. AB - The growing size of Canada's elderly population and its use of health care services has generated much discussion in policy circles and the popular press. With data from the National Population Health Survey, undertaken in 1994-95, the authors examine the health status of Canada's elderly population using 3 sets of measures: level of activity limitations, prevalence of chronic illnesses and self assessment of overall health. They also analyse the utilization of physician and institutional services. The profile of this population the authors develop is in many respects not much different from that of the remaining adult population, until the age of 75. People aged 75 and over are much more likely than other adults to have health problems and use health care services. Also, elderly women living alone and with low income are identified as an especially vulnerable group who need access to medical and nonmedical services if they are to remain in the community. Using Statistics Canada projection data the authors discuss some aspects of the elderly population's health status in the future. Their look into the future raises issues about the preparedness of health care providers and our health care system to meet the challenges of tomorrow's elderly population. PMID- 9347774 TI - Functional decline in old age. AB - Functional decline is a common condition, occurring each year in nearly 12% of Canadians 75 years of age and older. The model of functional health proposed by the World Health Organization (WHO) represents a useful theoretical framework and is the basis for the SMAF (Systeme de measure de l'autonomie fonctionelle or Functional Autonomy Measurement System), an instrument that measures functional autonomy. The functional decline syndrome, in which functional autonomy is diminished or lost, may present as an acute condition, i.e., a medical emergency for which the patient must be admitted to a geriatric assessment unit. The subacute form is a more insidious condition in which the patient requires comprehensive assessment and a rehabilitation program. A preventive approach based on screening of those at risk and early intervention should prevent or delay the appearance of functional decline or diminish its consequences. Effective strategies for the prevention of or rehabilitation from functional decline will help reduce the incidence of disabilities and the period of dependence near the end of life. These strategies are absolute prerequisites for controlling sociohealth expenses and, most importantly, for allowing people to live independently in old age. PMID- 9347775 TI - Alzheimer's disease: current knowledge, management and research. AB - Alzheimer's disease is a common neurological condition, appearing as early as age 40 but increasing dramatically in incidence over age 85. Different genetic factors are at play, modified by events over a lifetime. Clinical diagnosis is possible through careful history taking with a reliable informant and a minimum number of laboratory tests. A relatively predictable natural history can be observed, with progression through stages of cognitive loss, functional impairment and behavioural disinhibition or apathy. New medications such as donepezil offer hope for improving or stabilizing symptoms. Such treatment can be administered by primary care physicians with experience in the diagnosis and management of Alzheimer's disease. Disease stabilization, or even prevention, may be possible in the future. PMID- 9347776 TI - Depression in elderly medical inpatients: a meta-analysis of outcomes. AB - OBJECTIVE: To determine the prognosis of elderly medical inpatients with depression. DATA SOURCES: A MEDLINE search for relevant articles published from January 1980 to September 1996 and a search of the PSYCH INFO database for articles published from January 1984 to September 1996. The bibliographies of identified articles were searched for additional references. STUDY SELECTION: Eight reports (involving 265 patients with depression) met the following 5 inclusion criteria: original research, published in English or French, population of general medical inpatients, mean age of depressed patients 60 years and over, and affective state reported as an outcome. The validity of the studies was assessed according to the criteria for prognostic studies described by the Evidence-Based Medicine Working Group. DATA EXTRACTION: Information about the patient population, the proportion of cases detected and treated by attending physicians, the length of follow-up, the affective outcome and the prognostic factors was abstracted from each report. DATA SYNTHESIS: All of the studies had some methodologic limitations. A meta-analysis of outcomes at 3 months or less indicated that 18% of patients were well, 43% were depressed and 22% were dead. At 12 months or more, 19% were well, 29% were depressed and 53% were dead. Factors associated with worse outcomes included more severe depression, more serious physical illness and symptoms of depression before admission. CONCLUSIONS: Elderly medical inpatients who are depressed appear to have a very poor prognosis: the recovery rate among these patients is low and the mortality rate high. PMID- 9347779 TI - Medical management of frailty: confessions of a gnostic. AB - Geriatric Medicine is concerned chiefly with the care of frail elderly people, especially when they become ill. Physicians face special challenges in dealing with such patients, who tend to have multiple interacting medical and social problems, impaired function, altered pharmacokinetics and pharmacodynamics, atypical disease presentations and to be affected by polypharmacy. The joy of geriatrics is in systematically meeting each of these challenges, but the techniques that geriatricians use to do so must not be kept secret. More must be done to encourage all physicians to use these techniques in caring for frail elderly people who are ill. PMID- 9347777 TI - Pharmacologic treatment of depression in late life. AB - A number of age-related factors, including changes in pharmacokinetics and pharmacodynamics, medical comorbidity and an increased risk of drug-drug interaction, can complicate the pharmacologic management of depression in late life. Nevertheless, over 80% of elderly depressed patients will eventually respond to vigorous treatment and, when treated over 2 years, up to 75% of those will not have a relapse or recurrence of depression. This article reviews a number of issues relating to the pharmacotherapy of depression in elderly people. In particular, it discusses the similarities and differences between various antidepressant medications, issues pertaining to dosing and length of treatment, and management of the patient who does not respond to first-line treatment. The author emphasizes that, because of the high risk of relapse and recurrence, a long-term collaboration between the patient and the physician is required to successfully manage depression in late life. PMID- 9347780 TI - Detecting and managing elder abuse: challenges in primary care. The Research Subcommittee of the Elder Abuse and Self-Neglect Task Force of Hamilton Wentworth. AB - OBJECTIVE: To determine family physicians' perceptions of barriers and strategies in the effective detection and appropriate management of abused elderly people. DESIGN: Questionnaire survey; the protocol included an advance notification letter and 3 follow-up mailings. SETTING: Regional Municipality of Hamilton Wentworth, Ont. PARTICIPANTS: All active nonspecialist physicians who reported seeing elderly patients in their practices were eligible for inclusion. Fifty health service organization (HSO) physicians were randomly selected from among those listed with the HSO Mental Health Program, and 200 fee-for-service physicians were randomly selected from the Canadian Medical Directory. Of the 189 eligible physicians 122 returned completed questionnaires, a response rate of 65%. OUTCOME MEASURES: Physicians' ratings of the importance of potential barriers in assisting older people experiencing abuse and of the usefulness of strategies for dealing with elder abuse. RESULTS: Physicians identified the following barriers as fairly or very important: denial of abuse, resistance to intervention, not knowing where to call for help, lack of protocols to assess and respond to abuse, lack of guidelines about confidentiality, fear of reprisal, and lack of knowledge of the prevalence and definition of elder abuse. Strategies deemed to be helpful included a single agency to call, a directory of services, a list of resource people, an educational package, guidelines for detection and management, reimbursement for time spent on legal matters, continuing education, revision of fee structure and a central library of resources on elder abuse. CONCLUSION: Although the physicians perceived numerous barriers to their detection and management of elder abuse, they identified many strategies that could be implemented at a local level. Preparation of an algorithm to help physicians is the next phase of this work. PMID- 9347778 TI - Revisiting the O complex: urinary incontinence, delirium and polypharmacy in elderly patients. AB - Urinary incontinence, delirium and polypharmacy are common, challenging problems encountered in elderly patients. Review of the literature shows that these conditions are interrelated. For example, polypharmacy can lead to delirium, which, in turn, can lead to urinary incontinence. The drugs prescribed for urinary incontinence can precipitate delirium or contribute to polypharmacy. The underlying causes for these problems in elderly patients are frequently complex, and management in turn must often be multifactorial. The occurrence of these problems should lead to careful evaluation followed by thoughtful, responsive treatment. Brief updates are given with recommendations for management directed at primary care physicians. PMID- 9347782 TI - Promoting the health of senior citizens. AB - Canada is experiencing a dramatic increase in the number of older people in its population. Adopting strategies that involve physician actions, a societal approach and individual participation may substantially improve the health of senior citizens. This article presents ways to improve the quality of life and reduce the risk of premature death through manoeuvres that can be initiated by physicians in the context of the periodic health examination. The authors highlight the role of evidence in choosing the most appropriate interventions, speculate on areas of future importance and emphasize a societal approach to population health. PMID- 9347784 TI - Are we in store for some intergenerational warfare? PMID- 9347783 TI - Care for Canada's frail elderly population: fragmentation or integration? AB - Budget constraints, technological advances and a growing elderly population have resulted in major reforms in health care systems across Canada. This has led to fewer and smaller acute care hospitals and increasing pressure on the primary care and continuing care networks. The present system of care for the frail elderly, who are particularly vulnerable, is characterized by fragmentation of services, negative incentives and the absence of accountability. This is turn leads to the inappropriate and costly use of health and social services, particularly in acute care hospitals and long-term care institutions. Canada needs to develop a publicly managed community-based system of primary care to provide integrated care for the frail elderly. The authors describe such a model, which would have clinical and financial responsibility for the full range of health and social services required by this population. This model would represent a major challenge and change for the existing system. Demonstration projects are needed to evaluate its cost-effectiveness and address issues raised by its introduction. PMID- 9347781 TI - Caring for elderly people at home: the consequences to caregivers. AB - The emphasis on home-based care is one important aspect of health services restructuring initiatives in Canada. Fundamental to the preference for home-based care over institutional care is the expectation that family caregivers will be available in the home to support patients who would otherwise be in an institution. The authors explore the potential impact of this devolution of services from institutions to the home in 2 vulnerable patient populations- elderly patients with dementia and elderly patients with terminal illnesses. Community-based surveillance strategies are needed to determine the true health, quality-of-life and economic outcomes of these restructuring initiatives. PMID- 9347785 TI - Specialists are getting older, working longer. PMID- 9347786 TI - A practical guide for the diagnosis and treatment of acute sinusitis. AB - OBJECTIVE: To develop guidelines for the diagnosis and management of acute sinusitis. OPTIONS: Diagnostic clinical criteria and imaging techniques, the role of antimicrobial therapy and duration of treatment, and the role of adjunct therapy, including decongestants, glucocorticosteroids and nasal irrigation. OUTCOMES: Improved accuracy of clinical diagnosis, better utilization of imaging techniques and rational use of antimicrobial therapy. EVIDENCE: A MEDLINE search for relevant articles published from 1980 to 1996 using the MeSH terms "sinusitis," "acute sinusitis," "respiratory infections," "upper respiratory infections," "sinusitis" and "diagnosis," "sinusitis" and "therapy," "sinusitis" and "etiology," and "antimicrobial resistance" and search for additional articles from the reference lists of retrieved articles. Papers referring to chronic sinusitis, sinusitis in compromised patients and documented nonbacterial sinusitis were excluded. The evidence was evaluated by participants at the Canadian Sinusitis Symposium, field in Toronto on April 26-27, 1996. VALUES: A hierarchical evaluation of the strength of evidence modified from the methods of the Canadian Task Force on the Periodic Health Examination was used. Strategies were identified to deal with problems for which no adequate clinical data were available. Recommendations arrived at by consensus of the symposium participants were included. BENEFITS, HARMS AND COSTS: Increased awareness of acute sinusitis, accurate diagnosis and prompt treatment should reduce costs related to unnecessary investigations, time lost from work and complications due to inappropriate treatment. As well, physicians will be better able to decide which patients will not require antimicrobial therapy, thus saving the patient the cost and potential side effects of treatment. RECOMMENDATIONS: Clinical diagnosis can usually be made from the patient's history and findings on physical examination only. Five clinical findings comprising 3 symptoms (maxillary toothache, poor response to decongestants and a history of coloured nasal discharge) and 2 signs (purulent nasal secretion and abnormal transillumination result) are the best predictors of acute bacterial sinusitis (level I evidence). Transillumination is a useful technique in the hands of experienced personnel, but only negative findings are useful (level III evidence). Radiography is not warranted when the likelihood of acute sinusitis is high or low but is useful when the diagnosis is in doubt (level III evidence). First-line therapy should be a 10-day course of amoxicillin (trimethoprim-sulfamethoxazole should be given to patients allergic to penicillin) (level I evidence) and a decongestant (level III evidence). Patients allergic to amoxicillin and those not responding to first-line therapy should be switched to a second-line agent. As well, patients with recurrent episodes of acute sinusitis who have been assessed and found not to have anatomic anomalies may also benefit from second-line therapy (level III evidence). VALIDATION: The recommendations are based on consensus of Canadian and American experts in infectious diseases, microbiology, otolaryngology and family medicine. The guidelines were reviewed independently for the advisory committee by 2 external experts. Previous guidelines did not exist in Canada. PMID- 9347787 TI - HLA-DRB1*04 and DRB1*14 alleles are associated with susceptibility to pemphigus among Japanese. AB - It has previously been demonstrated that susceptibility to pemphigus vulgaris is associated with human leukocyte antigen (HLA)-DR4 serologic specificity among Ashkenase Jews, and with DR4 as well as DR6 (DR14) in other ethnic groups. We genotyped HLA-DRB1, DQA1, DQB1, and DPB1 alleles in 16 patients with pemphigus by polymerase chain reaction-restriction fragment length polymorphism, to find evidence of potential HLA class II allele associations with pemphigus in Japanese patients who have a relatively homogeneous ethnic background. All nine patients with pemphigus vulgaris and five of seven patients with pemphigus foliaceus carried one or two alleles of HLA-DRB1*04 (*0403, *0406) and HLA-DRB1*14 (*1401, *1405, *1406) subtypes. Sequence analysis of these DRB1*04 and DRB1*14 alleles revealed the amino acid homology of phenylalanine at position 26 and valine at position 86 with the DRB1*0402 allele that reportedly confers a strong susceptibility to pemphigus vulgaris in Ashkenazi Jews. Thus our findings, together with previous HLA studies on pemphigus vulgaris patients of different ethnic groups, suggest that HLA-DRB1*04 and DRB1*14 alleles are commonly associated with pemphigus vulgaris across racial barriers. These HLA-DRB1 alleles are likely to be also associated with pemphigus foliaceus. Further studies on more diverse ethnic populations will be helpful in determining the significance of the association between certain amino acid residues of the class II molecules and disease susceptibility to pemphigus vulgaris as well as pemphigus foliaceus. PMID- 9347789 TI - Despite the presence of UVB-induced DNA damage, HLA-DR+ cells from ex vivo UVB exposed human skin are able to migrate and show no impaired allostimulatory capacity. AB - In this study, we investigated the effect of ultraviolet B radiation on human Langerhans cell function. Normal human skin was irradiated ex vivo with single doses of ultraviolet B. For assessment of T-cell stimulatory function, cells that spontaneously migrated from epidermal sheets were used, whereas full-thickness skin biopsies were used to investigate alterations in migratory properties. The cells migrating from ultraviolet B-exposed epidermal sheets demonstrated a decrease in the percentage of HLA-DR positive Langerhans cells, as well as a reduced capacity to induce proliferation of allogeneic T cells, when compared with cells migrating from nonexposed sheets. When a correction was made for the decreased number of HLA-DR positive Langerhans cells migrating from ultraviolet B exposed epidermis, however, it appeared that the capacity to induce T-cell proliferation was identical for Langerhans cells migrating from ultraviolet B exposed and nonexposed epidermis. The presence of ultraviolet B-induced DNA damage could be demonstrated in the Langerhans cells from ultraviolet B-treated skin, indicating that the cells had received significant doses of ultraviolet B. As regards the effect of ultraviolet B on migratory properties of Langerhans cells, we found not only that reduced numbers of CD1a-positive Langerhans cells migrated from the ultraviolet B-exposed full-thickness skin, but also that there was a reduction in CD1a-positive Langerhans cells in the epidermis. This implies that ultraviolet B induces death of Langerhans cells as well as loss of cell surface molecules rather than altering Langerhans cells migration, whereas the Langerhans cells that were still able to migrate fully retained the capacity to activate allogeneic T cells. PMID- 9347788 TI - Epithelial-stromal interactions modulating penetration of matrigel membranes by HPV 16-immortalized keratinocytes. AB - The role of epithelial-stromal interactions in the progression of human papillomavirus-associated squamous intraepithelial lesions to invasive cervical cancer is poorly understood. Using the Matrigel artificial basement membrane assay as a model of keratinocyte invasion, the effects of selected growth factors on penetration of human papillomavirus 16-immortalized keratinocytes through Matrigel were studied. Also studied in this model were the effects of conditioned media from fibroblast lines derived from normal cervical tissues (normal fibroblasts) and adjacent cervical cancer biopsies (tumor-associated fibroblasts) and from primary keratinocytes. Addition of basic fibroblast growth factor, transforming growth factor-alpha, and hepatocyte growth factor/scatter factor or conditioned media from tumor-associated fibroblasts to the Matrigel resulted in near-doubling of penetration of human papillomavirus 16-immortalized keratinocytes, whereas transforming growth factor-beta, platelet derived growth factor-B, or conditioned media from primary keratinocytes decreased penetration 10-fold. Antibodies to basic fibroblast growth factor abrogated the stimulatory effects of conditioned media from tumor-associated fibroblasts on keratinocyte penetration, whereas antibodies to transforming growth factor-beta abrogated the inhibitory effects of conditioned media from normal fibroblasts on keratinocyte penetration. S1 nuclease protection and enzyme-linked immunosorbent assay showed increased expression of transforming growth factor-beta and decreased expression of basic fibroblast growth factor in normal compared with tumor-associated fibroblasts. Messenger RNA in situ hybridization of five cervical cancer biopsies demonstrated basic fibroblast growth factor expression in stromal cells surrounding nests of invading keratinocytes. Epithelial-stromal interactions mediated by growth factors such as transforming growth factor-beta and basic fibroblast growth factor modulate penetration of human papillomavirus 16 immortalized keratinocytes through Matrigel in vitro and these interactions may also be operative in vivo. PMID- 9347790 TI - Increased interstitial histamine concentration in the psoriatic plaque. AB - The psoriatic plaque contains an increased number of mast cells that are thought to have an important role in the initiation and maintenance of psoriatic lesions through the release of mediators such as histamine, proteoglycans, lipid mediators, and cytokines. It is not known, however, whether the interstitial concentration of histamine (and other mediators) is truly increased in the psoriatic plaque. The aim of the present study was to examine histamine concentration and histamine release from involved and uninvolved skin of psoriatic patients. Intracutaneous microdialysis was performed in lesional and nonlesional skin of 23 psoriatic subjects. The relative recovery of histamine was assessed after calibration in situ to approximately 76% in both lesional and nonlesional skin. The interstitial histamine concentration was 32 +/- 3 nmol per liter in lesional skin and 13 +/- 1 nmol per liter in nonlesional skin (mean +/- SEM) (p < 0.001). Dermal histamine release was estimated according to the Fick principle after measurements of the arterialized venous plasma histamine concentration (3 +/- 1 nmol per liter) and blood flow and was found to be 10-fold increased in lesional compared with nonlesional skin. The results are compatible with the hypothesis that mast cells in lesional skin secrete an increased amount of histamine that may contribute to the immunostimulation and inflammation in the psoriatic plaque. PMID- 9347791 TI - Most primary cutaneous CD30-positive lymphoproliferative disorders have a CD4 positive cytotoxic T-cell phenotype. AB - Primary cutaneous CD30-positive (anaplastic) large T-cell lymphomas and lymphomatoid papulosis are closely related types of cutaneous T-cell lymphoma with a favorable prognosis. The neoplastic T cells in these conditions have the phenotype of activated CD3+, CD4+, CD8-, CD30+ skin homing T cells, but their normal counterpart has not yet been defined. To further characterize the cellular origin of the neoplastic T cells, skin biopsies from 14 patients with primary cutaneous CD30-positive (anaplastic) large T-cell lymphomas, nine patients with lymphomatoid papulosis, and six patients with primary cutaneous CD30-negative pleomorphic large T-cell lymphomas were stained with monoclonal antibodies against cytotoxic cell-associated molecules granzyme B and T-cell restricted intracellular antigen. In nine of nine lymphomatoid papulosis and in 10 of 14 CD30-positive primary cutaneous large T-cell lymphomas, expression of granzyme B and T-cell restricted intracellular antigen by variable numbers of neoplastic cells was found. Expression of granzyme B by the neoplastic CD30-positive T cells was confirmed by double-staining for granzyme B and CD30 (three cases) and by the detection of granzyme B mRNA using RNA in situ hybridization (one case). In most cases equal numbers of granzyme-B-positive and T-cell restricted intracellular antigen positive tumor cells were observed. In five of six CD30-negative primary cutaneous large T-cell lymphomas the neoplastic cells did not express these proteins, whereas in one case a sporadic positive tumor cell was found. These results demonstrate that, in contrast to primary cutaneous CD30-negative pleomorphic large T-cell lymphomas, the neoplastic cells in most primary cutaneous CD30-positive (anaplastic) large T-cell lymphomas and lymphomatoid papulosis have a unique CD4+, CD8-, cytotoxic T-cell phenotype. PMID- 9347792 TI - Cultured human keratinocytes express tropoelastin. AB - We detected elastin mRNA in cultured normal human keratinocytes by RNase protection assay. The content of elastin mRNA was estimated at approximately one twentieth of that of cultured skin fibroblasts. Tropoelastin polypeptide with a molecular weight of 68 kDa was detected in the preparation of culture medium of normal human keratinocytes by western blot assays using anti-tropoelastin antibody. Immunohistochemical studies also demonstrated positive staining in cultured normal human keratinocytes as well as in skin fibroblasts. The expression of elastin by normal human keratinocytes was found to reach a maximum level at the quiescent phase of keratinocyte growth. When normal human keratinocytes were cultured on tropoelastin-coated dishes, their growth potential was greatly suppressed compared with other matrix protein-coated dishes. These results suggest that cultured normal human keratinocytes can actively synthesize elastin and that keratinocyte elastin may act as a growth-regulator for keratinocytes. PMID- 9347793 TI - Two Gq class G proteins are expressed in human keratinocytes. AB - G proteins link many cell surface receptor generated signals to activation of multiple cellular processes in all tissues. There is specificity in the receptor interaction with the G protein and in the interaction of the specific G protein with different effector molecules. The purpose of this study was to determine some of the biologically relevant G proteins in keratinocytes. The G alpha subunit of the heterotrimeric G protein was investigated because much of the biologic activity and the receptor specificity resides there. A polymerase chain reaction strategy was used that amplified multiple G alpha gene segments between conserved primer sites from keratinocyte first strand cDNA. Two Gq class G proteins, G alpha 16 and G alpha y, were identified. Using northern analysis and in situ hybridization, mRNA of both of these genes were detected in keratinocytes in culture and in epidermal keratinocytes. G alpha y was expressed in multiple other cell types and tissues, but G alpha 16 was restricted in expression to keratinocytes and keratinocyte-derived adnexal structures in the skin. G alpha 16 has previously been reported to be limited in expression to hematopoietic cells. The physiologic receptor to which it couples in neutrophils is reported to be a C5a receptor. The receptor to which it couples in keratinocytes has not been elucidated but by analogy may be another chemokine receptor. We hypothesize that G alpha 16 is an important conduit for responses to inflammatory signals in keratinocytes. PMID- 9347794 TI - Pemphigus IgG induces expression of urokinase plasminogen activator receptor on the cell surface of cultured keratinocytes. AB - We previously found that the binding of pemphigus IgG to desmogleins caused marked activation of phospholipase C, a transient increase in inositol 1,4,5 trisphosphate production, and a concomitant increase in the intracellular calcium concentration in DJM-1 cells, a squamous cell carcinoma line. The binding of pemphigus IgG to cell membranes increased the activity of urokinase plasminogen activator in culture medium and induced subsequent cell-cell detachment in DJM-1 cells. Because urokinase plasminogen activator activates the conversion of plasminogen to plasmin by binding to urokinase plasminogen activator receptor evading inhibitors in serum, it is likely that plasmin is generated only in microenvironments adjacent to urokinase plasminogen activator receptor on the cell surface. It is not known whether pemphigus IgG causes acantholysis by inducing urokinase plasminogen activator receptor expression on the cell surface and secreting urokinase plasminogen activator in inhibitor-rich environments. We examined the effects of pemphigus IgG on urokinase plasminogen activator receptor expression in DJM-1 cells and normal keratinocytes by immunoblot analysis and immunofluorescence microscopy using antibodies to urokinase plasminogen activator receptor. IgG were obtained from serum samples from eight patients with bullous pemphigoid, five patients with pemphigus vulgaris, seven patients with pemphigus foliaceus, and eight normal subjects. Pemphigus vulgaris and pemphigus foliaceus IgG significantly increased the urokinase plasminogen activator receptor expression on the surface of DJM-1 cells and normal keratinocytes after 3- and 7 d incubation compared with normal IgG. These results suggest that enhanced urokinase plasminogen activator activity and urokinase plasminogen activator receptor expression activates plasmin in the limited cell surface of pemphigus IgG-bound keratinocytes and may contribute to the pathogenesis of differential acantholysis in pemphigus vulgaris and pemphigus foliaceus. PMID- 9347795 TI - Clonality in nevocellular nevus and melanoma: an expression-based clonality analysis at the X-linked genes by polymerase chain reaction. AB - The true nature of nevocellular nevus is still unknown and it has been ambiguously classified as a neoplasm or a hamartoma. We studied the clonality of nevocellular nevus and melanoma (malignant melanoma), using an expression-based clonality analysis at the X-linked genes by means of polymerase chain reaction. DNA was extracted from cryostat sections of 20 nevocellular nevi (10 compound and 10 intradermal type) and five melanomas from female patients. A polymorphic portion of the inactivated X-linked gene was amplified after selective digestion of the active X-chromosome with a methylation-sensitive restriction enzyme, Hpa II. Paternal- and maternal-derived fragments were resolved with electrophoresis using the polymorphic restriction endonuclease (BstX I) site for the phosphoglycerate kinase assay, and using the difference of CAG repeats for the human androgen-receptor gene assay. Both assays revealed that all informative nevocellular nevi were polyclonal in origin and all melanomas were monoclonal. Results of the clonality were independent of either the histologic type of nevocellular nevus or whether the nevocellular nevus was of congenital or acquired origin. Thus, nevocellular nevus, congenital or acquired, may be a hamartomatous rather than a neoplastic lesion. The analysis of clonality could be applied to the differential diagnosis of benign melanocytic disease and melanomas. PMID- 9347796 TI - No evidence for involvement of the growth hormone/insulin-like growth factor-1 axis in psoriasis. AB - We have examined whether alterations in the growth hormone/insulin-like growth factor-1 axis play a role in the pathogenesis of psoriasis. Serum, urine, full skin biopsies, and suction blister roofs were obtained from patients with psoriasis and from healthy controls. Serum concentrations of insulin-like growth factor-1 and insulin-like growth factor binding protein-3 were measured by radioimmunoassay. Growth hormone-binding protein was measured by ligand-mediated immunofunctional assay. Growth hormone concentration in urine was measured by an immunometric assay, and growth hormone receptor-gene expression was measured by RNase protection assay or by quantitative reverse transcriptase polymerase chain reaction in total RNA isolated from epidermal suction blister roofs. Serum concentrations of insulin-like growth factor-1 (249 +/- 12 micrograms per liter, mean +/- SEM, n = 42, and 277 +/- 21 micrograms per liter, n = 9, for psoriatic patients and controls, respectively), insulin-like growth factor binding protein 3 (3.1 +/- 0.08 mg per liter, n = 42, and 3.3 +/- 0.22 mg per liter, n = 9), growth hormone-binding protein (344 +/- 65 pmol per liter, n = 10, and 311 +/- 83 pmol per liter, n = 9), urinary growth hormone excretion during 24 h (12.8 +/- 2.7 microIU per 24 h, n = 12, and 12.3 +/- 1.6 microIU per 24 h, n = 9), and epidermal growth hormone receptor gene expression [32 +/- 12 x 10(3) mRNA transcripts per microgram total RNA (involved skin), n = 11, and 47 +/- 14 x 10(3) mRNA transcripts per microgram total RNA, n = 9] were similar in patients and controls. For insulin-like growth factor-1 and insulin-like growth factor binding protein-3 the values in psoriatic patients were also similar to those in larger control groups, n = 195 and n = 400, respectively. In addition, we found no evidence of local expression of growth hormone or prolactin in full skin punch biopsies from psoriatic involved skin by reverse transcriptase polymerase chain reaction. In conclusion, our results suggest that alterations in the growth hormone/ insulin-like growth factor-1 axis do not play a major role in the pathogenesis of psoriasis. PMID- 9347798 TI - Expression of two Ig family adhesion molecules in the murine hair cycle: DCC in the bulge epithelia and NCAM in the follicular papilla. AB - The hair cycle involves remodeling of cells and of cell groups into a complex follicular structure. During skin appendage development, adhesion molecules such as neural cell adhesion molecule (NCAM) and deleted in colon carcinoma (DC) participate in the formation of cell groups. NCAM has been found to be expressed in the mesenchyme during mouse hair follicle induction. DCC expression has been observed in the epithelial cells of the developing feather. We postulate that these two molecules may also define cell groups in the cycling hair follicle. Here we report their spatio-temporal expression patterns during the depilation induced murine hair cycle. NCAM expression was also examined in positive and negative hair-inductive follicular papilla cell lines. Throughout the hair cycle, DCC expression was confined to the basal keratinocytes of the epidermis and the epithelial portion of the hair follicle. During mid-anagen, two types of deleted in colon carcinoma staining were observed. One was a cell surface pattern seen in the epithelial cells in the bulge region where the follicular stem cells reside. The other was a diffuse cytoplasmic staining pattern in the transient hair follicle epithelia located below the bulge region. Prominent NCAM staining was observed in the follicular papilla throughout the hair cycle and was accompanied by weak staining of the matrix epithelia. NCAM expression correlated with hair induction by a follicular papilla cell line. The results suggest that DCC and NCAM define the permanent cell groups of the hair follicle and that NCAM is important for hair induction. PMID- 9347797 TI - Linkage studies in erythrokeratodermias: fine mapping, genetic heterogeneity and analysis of candidate genes. AB - Erythrokeratodermias are a clinically heterogeneous group of rare autosomal dominant disorders of cornification with overlapping features including hyperkeratosis and erythema. We ascertained five extended pedigrees with different phenotypes for a linkage study. Three families presented with localized erythrokeratodermia variabilis, and one with erythrokeratodermia and ataxia. Another family had Greither disease associated with variable hyperkeratotic plaques. Despite their phenotypic differences, both erythrokeratodermia variabilis and erythrokeratodermia with ataxia map to a common region in 1p34 p35. Multipoint linkage and haplotype analyses place erythrokeratodermia variabilis between the marker D1S496 and D1S186 with a maximum LOD score of 12.88. Our linkage results provide compelling evidence for genetic homogeneity among families of mixed European and French-Canadian origin. In contrast, results excluded Greither's disease from the established erythrokeratodermia variabilis gene region indicating genetic heterogeneity of erythrokeratodermias. Based on recombinations, two genes assigned to 1p34-p35 were excluded: cartilage matrix protein and avian myelocytosis viral oncogene. Connexin-37 (GJA4), a member of the connexin gene family, maps within the erythrokeratodermia variabilis region and is an attractive candidate gene. Direct sequencing of the coding region of GJA4 in four patients revealed several variations, including a novel polymorphism within the 5' cytoplasmic domain, but no pathogenic mutations were found, thus excluding Connexin-37 as a candidate. There is evidence, however, that other epidermally expressed connexins cluster in this region, and one may yet be determined to play a role in the pathogenesis of erythrokeratodermia variabilis. PMID- 9347799 TI - A highly sensitive enzyme-linked immunosorbent assay for the detection of circulating anti-BP180 autoantibodies in patients with bullous pemphigoid. AB - The BP180 antigen, a component of the epidermal anchoring complex, has been identified as one of the major antigenic targets of autoantibodies associated with the blistering skin disease, bullous pemphigoid. Our research group has recently demonstrated that reactivity of bullous pemphigoid autoantibodies to the BP180 ectodomain is almost entirely restricted to a set of four antigenic sites clustered within the membrane-proximal noncollagenous stretch (NC16A). Using a passive transfer mouse model, antibodies to the corresponding noncollagenous region of murine BP180 were shown to trigger an inflammatory subepidermal blistering disease that closely mimics bullous pemphigoid. We now report the development of an enzyme-linked immunoabsorbent assay system that is extremely sensitive in detecting disease-specific autoantibodies in the sera of bullous pemphigoid patients. The target antigen in this assay is a recombinant form of the BP180 NC16A domain that contains all four of the well-defined bullous pemphigoid-associated antigenic sites. Of 50 randomly selected bullous pemphigoid sera tested, 47 (94%) were positive in this assay, whereas no specific reactivity was detected in any of the 107 controls. Interestingly, all three of the bullous pemphigoid sera that were negative in this assay had been obtained from patients who were already undergoing treatment. The NC16A enzyme-linked immunosorbent assay is more sensitive than any of the standard techniques for detecting circulating bullous pemphigoid autoantibodies, including other enzyme-linked immunosorbent assays, immunoblotting, and indirect immunofluorescence. Finally, the NC16A enzyme-linked immunosorbent assay provides immunologic information that cannot be obtained from direct immunofluorescence studies of skin biopsies, and that may well be relevant in the diagnosis and treatment of bullous pemphigoid. PMID- 9347800 TI - Novel glycine substitution mutations in COL7A1 reveal that the Pasini and Cockayne-Touraine variants of dominant dystrophic epidermolysis bullosa are allelic. AB - Mutations in the type VII collagen gene (COL7A1) have been shown to underlie dystrophic epidermolysis bullosa (DEB). The dominantly inherited forms of DEB have been divided into two clinical subcategories, the Pasini (DDEB-P) and the Cockayne-Touraine (DDEB-CT) variants, on the basis of the presence or absence of albopapuloid lesions. In this study, we have examined the molecular basis of DDEB in two Japanese families, one with DDEB-P and the other with DDEB-CT. Mutation detection strategy consisted of polymerase chain reaction amplification of COL7A1 from genomic DNA, followed by heteroduplex analysis and direct nucleotide sequencing. The results revealed heterozygous glycine substitution mutations, G2076D and G2034R, in these families, respectively. Thus, these two variants of DDEB are allelic, and subtle differences in the clinical presentation may reflect the precise position of the mutation along the type VII collagen molecule. Alternatively, the nature of the substituting amino acid (D versus R) may influence the clinical phenotype. This is the first demonstration of a COL7A1 mutation in DDEB-P, and brings the total number of dominant DEB variants with underlying glycine substitutions in COL7A1 to five, including the pretibial and localized variants as well as the Bart's syndrome, in addition to DDEB-P and DDEB CT. PMID- 9347801 TI - Erythropoietic protoporphyria: four novel frameshift mutations in the ferrochelatase gene. AB - Human erythropoietic protoporphyria is an inherited disorder of the heme metabolic pathway caused by defects in the gene for ferrochelatase, the terminal enzyme of the pathway that catalyzes chelation of ferrous iron into protoporphyrin IX to form heme. Mutation analysis was performed for families with erythropoietic protoporphyria and four novel frameshift mutations were identified. Two of the mutations, 205insA and 215insT in exon 3 of the ferrochelatase gene, are single bp insertions. The other two, 400delA in exon 4 and 678delG in exon 6, are single bp deletions. All of the mutations result in premature termination codons downstream shortly after the mutation sites, and in one case the premature termination codon caused by 400delA was also shown to reduce mRNA level via nonsense-mediated mRNA decay. PMID- 9347802 TI - An alanine to proline mutation in the 1A rod domain of the keratin 10 chain in epidermolytic hyperkeratosis. AB - We report a mutation in a case of epidermolytic hyperkeratosis that results in a proline for alanine substitution in the residue position 12 of the 1A subdomain of the keratin 10 chain (codon 158). The disease phenotype is consistent with the inappropriate substitution of a proline near the beginning of the rod domain, because it is likely to seriously disrupt the structural organization of coiled coil molecules within keratin intermediate filaments. Mutations/substitutions in this position have not been reported in any keratin disease. Position 12 is an alanine in all intermediate filament chains, and lies in the outer b heptad position of the coiled-coil. In vitro peptide interference assembly assays revealed that substitutions that alter residue size or charge at this position primarily interfere with keratin filament elongation. PMID- 9347803 TI - Significantly earlier age at onset for the HLA-Cw6-positive than for the Cw6 negative psoriatic sibling. PMID- 9347804 TI - Another support for the location of epidermal stem cells residing adjacent to the tips of dermal papillae in the interfollicular epidermis. PMID- 9347805 TI - Reducing cardiac surgical trauma: the minimally invasive direct coronary artery bypass. AB - BACKGROUND: The concept of minimal surgical trauma is revolutionizing many surgical subspecialties, including cardiac surgery. Coronary artery revascularization can now be accomplished either thoracoscopically or through a small thoracotomy, sternotomy, or epigastric incision, with or without cardiopulmonary bypass (CPB). METHODS: The current literature was reviewed with regard to patient selection criteria for coronary artery bypass grafting (CABG) without CPB, indications for minimally invasive direct coronary artery bypass (MIDCAB), surgical and anesthetic technique, and outcome. RESULTS: The MIDCAB is largely used in cases of single or double vessel disease. The procedure is done either thoracoscopically or under direct vision through a small incision rather than standard sternotomy. In non-CPB cases, the heart is pharmacologically manipulated to create a quiet operative field. Patients may be extubated and become ambulatory shortly after surgery and be discharged within a few days. CONCLUSIONS: The MIDCAB avoids median sternotomy and, in many cases, CPB. MIDCAB may prove to play a prominent role in management of coronary artery disease in the future. PMID- 9347806 TI - Management of the adnexal mass in the 1990s. AB - BACKGROUND: Ovarian carcinoma is the leading cause of deaths from female genital cancers in the United States. During the last three decades, advances in diagnostic techniques, surgical techniques, and adjuvant chemotherapy have led to improved survival in some patients who have an adnexal mass that is later diagnosed as malignant. METHODS: A review of the current technique, compiled with our changing management, was done to help identify possible pitfalls in the initial management of the adnexal mass in specific age groups. The expensive and controversial issues such as screening, management of patients with a genetic history, and management with laparoscopy were reviewed. RESULTS: Appropriate initial surgery improves survival in patients with adnexal masses, later determined to be malignant, particularly when adjuvant, modern combination chemotherapy is used. Laparoscopy for suspicious adnexal masses cannot be condoned, unless immediate appropriate surgical staging can be done. CONCLUSIONS: Awareness and implementation of current diagnostic and treatment modalities can improve survival in the patient with an adnexal mass that is later found to be malignant. PMID- 9347807 TI - Nephron-sparing surgery compared with radical nephrectomy for renal tumors: current indications and results. AB - BACKGROUND: Small asymptomatic solid renal masses are being found more often through the frequent use of abdominal ultrasonography and computed tomography. Nephron-sparing renal surgery is being done more often to treat these small lesions. A retrospective review was done to determine the effectiveness of this treatment. METHODS: Patients who had nephron-sparing renal surgery (group 1-35 patients) were compared with those who had radical nephrectomy (group 2-71 patients) for renal cell carcinoma smaller than 5 cm. RESULTS: The two groups had only small differences in fall in hematocrit, transfusion rates, operative time, and hospital stay. Major surgical complications were more frequent in group 1. After a median follow-up of 3.1 years, there has been no recurrence of tumor and there were no surgery-related or cancer-related deaths in either group. CONCLUSION: Nephron-sparing renal surgery appears to be a safe and effective alternative to radical nephrectomy for localized small renal tumors. PMID- 9347808 TI - Cancer incidence among predominantly black, rural-poor populations in southern states. AB - BACKGROUND: A seven-county, predominantly black, rural-poor population in Alabama is targeted for a program aimed at improving access to state-of-the-art cancer care. This paper presents combined age-adjusted cancer incidence rates for predominantly black, rural counties in North Carolina and Georgia similar to the Alabama counties and compares these rates with Surveillance, Epidemiology, and End Results (SEER) incidence rates. METHODS: Cancer incidence data from 1990 to 1993 were obtained from the Georgia Center for Cancer Statistics for 10 rural counties with predominantly black populations. Likewise, cancer incidence data from 1990 to 1993 were obtained for seven rural-poor counties in North Carolina from the North Carolina Central Cancer Registry. SEER incidence rates from 1990 to 1992 were obtained for nine SEER sites. RESULTS: The overall cancer incidence rate from North Carolina and Georgia is lower by 22% than the SEER rate. Cancer incidence rates for cancers of the breast, colon/rectum, lung, and prostate were at least 15% lower than the SEER rates, while the invasive cervical cancer rate was 1.78 times higher than the SEER rate. CONCLUSION: Blacks comprise about 50% of the population in these counties. In contrast, the SEER population is predominantly white, and the black population is primarily urban. Estimates of the number of cancer cases in black, rural-poor populations based on SEER incidence rates is not reflective of the cancer experience in these populations. PMID- 9347810 TI - Exposure to reduced sulfur gases impairs neurobehavioral function. AB - BACKGROUND: In the 19th century, deaths from acute exposure to hydrogen sulfide (H2S) portended permanent brain injury from nonlethal doses. The neurobehavioral effects of H2S exposures lasting from moments to years were compared in 16 subjects, 2 years to 22 years afterward. METHODS: Neurophysiologic and psychologic tests were used to appraise mood status and frequencies of 35 symptoms. Functions and frequencies, described as percent predicted adjusted for age, sex, educational achievement, and other factors, were compared with those in an unexposed population. RESULTS: Frequencies were elevated for 31 of 33 symptoms. Balance was impaired (246% predicted with eyes closed, 159% predicted with eyes open), and simple and choice reaction times were prolonged (151% and 130% predicted, respectively). Visual fields performance was decreased to 72% predicted (right) and 55% predicted (left), color discrimination was abnormal, and hearing was decreased. Psychologic domains showed cognitive disability, reduced perceptual motor speed, impaired verbal recall and remote memory, and abnormal mood status. CONCLUSIONS: Exposure to H2S must be avoided. PMID- 9347811 TI - Psychiatric perspective of pediatric human immunodeficiency virus infection. AB - BACKGROUND: Infection due to human immunodeficiency virus (HIV) has become a chronic disease of childhood, with increasing rates among adolescents and longer survival of those infected. This illness and its victims present a continuing challenge to the medical community. METHODS: We used computerized literature searches (MEDLINE and AIDSline) to identify research and review papers from medical, psychiatric, and psychology journals; we obtained statistics directly from the Centers for Disease Control and Prevention, National AIDS Clearinghouse. RESULTS: This overview of the topic includes epidemiology, transmission, diagnosis, psychiatric and neuropsychiatric manifestations, intervention, and the impact on families, caretakers, and health care workers. CONCLUSIONS: To psychiatrists, this disease presents the epitome of the biopsychosocial model, encompassing the biology of a viral disease with psychiatric manifestations, complicated by the societal place in which it has become entrenched. PMID- 9347813 TI - Acute colonic pseudo-obstruction complicated by cecal perforation in a patient with Parkinson's disease. AB - Acute colonic pseudo-obstruction (Ogilvie's syndrome) is characterized by physical examination and radiologic findings indicative of mechanical obstruction but in which no physical obstructive process can be found. Many factors have been associated with this syndrome which include electrolyte imbalance, systemic infection, drugs, and occasionally, neurologic disease. Reported here is a case of acute colonic pseudo-obstruction which developed in a patient with known Parkinson's disease and was complicated by cecal perforation, yet had a favorable outcome. PMID- 9347812 TI - Pediatric Graves' disease: therapeutic options and experience with radioiodine at the University of Mississippi Medical Center. AB - BACKGROUND: Pediatric Graves' disease can be life-threatening, and it adversely alters growth and development. Controversies concerning optimal therapy led us to review our 40 pediatric patients treated for Graves' disease from 1988 to 1996 to assess efficiency, efficacy, and safety of current therapy options. METHODS: Diagnosis of Graves' disease required clinical hyperthyroidism with supportive laboratory studies. Patients were given informed choices of therapy, which divided them into three groups. RESULTS: In group 1, 17 patients received antithyroid medications for 0.3 years to 6.0 years. Three required surgical thyroidectomy. Remissions (with or without thyroxine therapy) were achieved after 2 years to 5 years in 11 (65%). In group 2, 15 patients received antithyroid medications for 0.3 years to 5.0 years before receiving radioactive iodine (131I). One also required surgical thyroidectomy. Remissions were achieved after 1 year to 5 years in 10 (67%). In group 3, eight patients received initial 131I therapy. Remissions were achieved within 1 year in 7 (88%). CONCLUSIONS: Our results agree with and expand upon published reports on Graves' disease. Our data support early use of 131I as efficient, effective, and safe therapy for pediatric Graves' disease. PMID- 9347809 TI - Presentation and treatment of spontaneous aortocaval fistula. AB - BACKGROUND: Spontaneous rupture of abdominal aortic aneurysm into the inferior vena cava is rare. The clinical presentation is highly variable, and the diagnosis can be difficult, often being made only at operation. The aortocaval fistula results in a large left-to-right shunt, which can cause cardiac failure. Once the diagnosis is made, treatment is by surgical closure of the fistula and repair of the aneurysm with a graft. METHODS: This is a retrospective review of a single surgeon's experience with aortocaval fistula complicating abdominal aortic aneurysms. RESULTS: Over a 15-year period, we had five patients with spontaneous aortocaval fistula who were treated operatively. Preoperative diagnosis was made in two, suspected in one, and not made in two, one of whom died (the only perioperative death in the series). CONCLUSIONS: Spontaneous aortocaval fistulas are uncommon, and their preoperative recognition is difficult. Hematuria in association with an abdominal aortic aneurysm should raise the suspicion of an aortocaval fistula. Surgical correction is possible, with survival rates comparable to those associated with rupture of aneurysms into the retroperitoneum. Early operative control of the fistula is important to optimize the preload to the heart. PMID- 9347814 TI - Occult thyroid carcinoma manifested as a cystic neck mass. AB - Papillary carcinoma of the thyroid is occasionally manifested by palpable cervical adenopathy and an occult primary lesion. Isolated cystic metastases as the presenting sign are an uncommon finding. In the case reported, a cystic lesion in the patient's neck laterally proved to be an occult papillary carcinoma. PMID- 9347815 TI - An unusual cause of cyanosis in a patient with a carcinoid tumor. AB - We report the second case of a right-to-left interatrial shunt due to a carcinoid tumor, which was successfully corrected by surgery. The patient had closure of a patent foramen ovale and tricuspid valve replacement. One and a half years after surgery, the patient had no symptoms related to heart disease. PMID- 9347816 TI - Elemental mercurial poisoning. AB - A 39-year-old man injected 40 mL of elemental mercury in an attempted suicide 3 years before coming to our facility. No specific treatment regimen had been done since then. Chest x-ray films showed mercury deposits in the lungs, as well as around the injection site. The mercury concentration in his blood was at 96.3 micrograms (0.480 nmol/L), thus significantly elevated (given a reference range of up to 2 micrograms Hg/L), as was the renal mercury elimination. Despite mercurial deposits within the pulmonary circulation, the pulmonary function showed normal values, with no reduction of the diffusion capacity. There were signs of polyneuropathy. The patient was given sodium dimercaptopropanesulfate (Dimaval) for mercury complexation. This case report outlines the diagnosis and therapy for mercurial poisoning through metallic mercury. PMID- 9347818 TI - Vibrio vulnificus infection: an important cause of septicemia in patients with cirrhosis. AB - Vibrio vulnificus, a virulent gram-negative organism, is a normal inhabitant of coastal waters, including the Gulf of Mexico. Vibrio vulnificus infection has been recognized as a cause of fatal septicemia in chronically ill patients, particularly those with chronic liver disease. We report the case of a patient with chronic liver disease who had V vulnificus septicemia 2 days after eating raw oysters harvested in the Gulf Coast. Vibrio vulnificus septicemia should be suspected in all patients with underlying medical illnesses, particularly cirrhosis, who present with a febrile illness days after eating seafood or being exposed to saltwater. Physicians should advise their patients with cirrhosis and other chronic debilitating illnesses not to eat raw or undercooked seafood. PMID- 9347817 TI - Intermittent fever and pancytopenia in a young Mexican man. AB - We report a case of brucellosis in a young Mexican man who had weight loss, fever, and nausea. Physical examination revealed hepatosplenomegaly, and examination of the blood showed pancytopenia. This case illustrates the need for a high index of suspicion when patients living in the southern United States have these symptoms. PMID- 9347819 TI - Fatal iron intoxication in an infant. AB - Iron ingestion continues to be one of the major contributors to pediatric poisoning deaths. In 1995, more than 22,000 children unintentionally received preparations containing iron. Despite child-resistant packaging and education of both the public and medical personnel, the number of deaths has not significantly declined. Our patient, a 13-month-old child, ingested prenatal vitamins and despite aggressive efforts died 13 hours after his initial presentation. The patient's 3-year-old sibling had been evaluated at a local hospital 12 hours earlier for iron ingestion. Although there have been previous reports of death from iron ingestion, this case report is important because of the sibling's presentation and medical evaluation 12 hours before our patient's presentation. This case illustrates the need for physicians to inquire about other children in the home, possibly preventing further tragic outcomes. PMID- 9347821 TI - Dystonia associated with crack cocaine use. AB - Cocaine is a substance that has significant central stimulant action in the central nervous system. As cocaine abuse spreads throughout society, many neurologic side effects are appearing with increasing frequency. These side effects include seizures, tremor, focal neurologic deficits, headache, and dizziness. Recently, there have been reports of movement disorders associated with cocaine use. Cocaine use increases the incidence of acute dystonic reactions in patients being treated with dopamine blocking agents. There have also been rare reports of cocaine causing dystonia in patients who were taking no other street drugs or medications. Our report describes the case of a patient who had an acute dystonic reaction 12 hours after a crack cocaine binge. PMID- 9347822 TI - Surreptitious superwarfarin poisoning with brodifacoum. AB - Because of the emergence of warfarin resistance in rodents, second-generation anticoagulants named "superwarfarins" were developed and marketed in over-the counter rodenticide products. The availability of these compounds has resulted in accidental or intentional human ingestions, which cause severe bleeding. The methods for diagnosis and treatment of patients using superwarfarins are different from those for patients taking the regular warfarins. We report a case of intentional superwarfarin ingestion that caused petechiae and hematuria. Although the patient denied taking anticoagulant, the persistence of vitamin K dependent factor deficiency led us to investigate the serum for anticoagulant rodenticides. We found high levels of brodifacoum, a superwarfarin compound. This case emphasizes the need for suspicion of superwarfarin poisoning in patients who show unexplained bleeding due to deficiency of vitamin K-dependent factors and resistance to treatment. PMID- 9347823 TI - Granuloma annulare associated with malignancy. AB - Granuloma annulare is a benign dermatosis. Although granuloma annulare usually occurs as an idiopathic condition, it has been described in oncology patients. I report the case of a man in whom the appearance and resolution of granuloma annulare occurred in a paraneoplastic manner. The diagnosis of his previously unsuspected pulmonary adenocarcinoma was preceded by the concurrent onset of granuloma annulare lesions and tumor-related systemic symptoms. Two other patients with malignancy-associated granuloma annulare are discussed: a woman with breast cancer and a woman with cervical cancer. In these cases, the appearance of granuloma annulare was also temporally associated with the detection of a previously undiagnosed malignancy or the discovery of recurrent metastatic disease. The resolution of the dermatosis was temporally associated with the successful treatment of the neoplasm in these individuals. Granuloma annulare should be added to the list of cutaneous paraneoplastic syndromes that may occur in patients with solid tumors. PMID- 9347820 TI - CD8+ lymphocytic pneumonitis in a patient receiving cyproterone acetate. AB - A large number of drugs can induce pulmonary disease. We report the case of a female patient who was receiving cyproterone acetate for severe hirsutism. After 4 months of cyproterone therapy, she had dyspnea, pulmonary infiltrates, and a restrictive ventilatory defect. Bronchoalveolar lavage showed CD8+ lymphocytosis as well as increased neutrophils and eosinophils, a profile highly suggestive of an iatrogenic process. The patient showed no other significant clinical or biologic abnormality. Symptoms and functional abnormalities disappeared after withdrawal of cyproterone and reappeared after its reintroduction. This suggests that cyproterone acetate, a substance used to treat hirsutism and prostate cancer and in the composition of certain oral contraceptives, can be added to the list of drugs that may cause lymphocytic pneumonitis. PMID- 9347824 TI - Encephalopathy due to capillary leak syndrome. AB - Systemic capillary leak syndrome (SCLS) is characterized by intermittent attacks of leakage of intravascular fluids into the extravascular space. Hypovolemia, hemoconcentration, weakness, edema, and visceral congestion are resulting manifestations of SCLS. Most patients with SCLS have clear mentation during attacks, and encephalopathy is not a known manifestation of the syndrome. We report a patient with acute idiopathic capillary leak syndrome manifested in an acute encephalopathy. The possibility of SCLS should be considered in patients who have an encephalopathy and hemoconcentration. PMID- 9347825 TI - Breast metastasis from non-small cell lung cancer. AB - Breast is an unusual site for metastatic disease, particularly for non-small cell lung cancer. We report an unusual case of metastatic breast lesions from a primary anaplastic lung tumor and discuss the common and uncommon sites of metastasis from lung carcinomas. PMID- 9347826 TI - Surgery of the esophagus. Anatomy and physiology. AB - The esophagus can be divided into three parts: cervical, thoracic, and abdominal. Its blood supply, lymphatic drainage, innervation, and architecture of the esophageal wall are described. The topographic relationships of the esophagus and the gastroesophageal junction with neighboring structures are illustrated from the right and left thoroscopic and the laparoscopic viewpoints. Functionally, the esophagus consists of the upper esophageal sphincter; the esophageal body; and the lower esophageal sphincter. Their coordinated muscular activity transports the food bolus into the stomach, while maintaining a barrier against reflux of esophageal contents into the pharynx and gastric juice into the esophagus. PMID- 9347827 TI - Pathophysiology and endoscopic/balloon treatment of esophageal motility disorders. AB - Diagnostic and therapeutic dilemmas associated with esophageal dysmotility syndromes continue to confront physicians managing these patient populations. Although modern manometric systems have allowed us to better define normal parameters of esophageal motility, with the exception of primary achalasia, the clinical relevance of many aberrant motor patterns remains unclear. The novel use of botulinum toxin in idiopathic achalasia stems from increased understanding of the pathogenesis of the disease. Similarly, as our knowledge of the pathophysiology of other esophageal motor disorders grows, in conjunction with improved diagnostic capabilities, more effective management strategies may be used in the future. PMID- 9347828 TI - Surgical management of achalasia. AB - Although achalasia is not a common illness in the United States and Europe, there continues to be a need for surgical therapy for treatment. Laparoscopic Heller myotomy and partial fundoplication has, for the most part, replaced open surgery (abdominal or thoracic) as the surgical treatment of choice. In order to perform this procedure well, one must select patients carefully, evaluate them fully, and adhere to the technical principles required to achieve consistently good results. PMID- 9347829 TI - Basic considerations in gastroesophageal reflux disease. AB - The cause of foregut symptoms is often quite uncertain until a comprehensive evaluation has been performed. The critical elements of this evaluation include historic, radiographic, endoscopic, and physiologic data, and most importantly, the insight of a mature diagnostician. Patients who are not evaluated in a comprehensive way are at risk for serious postoperative problems; surgeons who perform interventions without appropriate diagnostic support may have to deal with these unhappy patients. In the long run, a complete workup provides the guidance for treatment and is cost-effective. PMID- 9347830 TI - Medical therapy of gastroesophageal reflux and management of esophageal strictures. AB - The goals of modern medical therapy for gastroesophageal reflux disease are threefold: first, eliminate symptoms; second, heal injured esophageal mucosa; third, manage and/or prevent complications. Selection of a particular medical regimen depends on the severity of the disease, effectiveness of the therapy, cost, and convenience of the medical regimen. An accurate diagnosis needs to be made in patients suspected with esophageal strictures. If there is a treatable underlying disease, specific therapy is essential. The goal of dilation therapy should be established and set about to accomplish in a timely, but unhurried fashion. Fluoroscopy and wire-guided dilators should be used liberally, especially for difficult strictures. PMID- 9347831 TI - Surgical treatment of gastroesophageal reflux disease. AB - Modern techniques have substantially improved the outcome of surgical therapy for reflux. Surgery, therefore, should not be considered a method of last resort but instead a reasonable alternative to treat patients with abnormal reflux. Adequate preoperative staging of the disease helps design the most appropriate operation. Minimally invasive techniques improve exposure and enhance recovery. Control of symptoms is achieved in the great majority of patients, and complications are minimal if the operation is performed following basic principles of sound operative technique. PMID- 9347832 TI - Management of the failed antireflux operation. AB - A further operation is required in a small proportion of patients who have had prior antireflux surgery. This has a surprisingly good chance for success in appropriately evaluated patients. The surgeon must make very specific decisions regarding the surgical approach. The use of laparoscopy for redo surgery is being defined. PMID- 9347833 TI - Esophageal replacement for end-stage benign esophageal disease. AB - The fact that esophageal resection and foregut reconstruction for benign disease can be performed with only a 2% mortality and minimal morbidity is encouraging news to patients who are crippled by the various manifestations of end-stage disease. The continuation of slow, anxious, and socially restricted alimentation or the maintenance of nutrition by enteral or parenteral means is unnecessary. The patient should be referred to a unit skilled in evaluating foregut function, performing esophageal replacement surgery, and caring for patients in the perioperative period. In our experience, the colon, when available, is the preferred conduit for esophageal replacement over the long term. Even though some subtle preoperative symptoms of foregut dysfunction may persist after surgery, the overall outcome is generally judged to be satisfactory. Indeed, patients can re-enter society and live a normal and fulfilled life after remedial surgery. Prolonged attempts at medical management of patients with severe derangements of esophageal structure and function are not warranted. Long-term esophageal replacement for severe end-stage benign disease can be accomplished with low mortality, a high degree of success, and a marked improvement in the quality of alimentation. Reconstruction restores the pleasure of eating and is viewed by the patient to be highly successful. PMID- 9347835 TI - Biology of esophageal cancer and the role of combined modality therapy. AB - 1. The biology of esophageal cancer involves multifactorial environmental and genetic events. 2. The understanding of the clinical significance of molecular markers is rapidly evolving. 3. Combined-modality approaches should still include surgery in good performance status (ECOG scale < or = 2) patients. 4. Neoadjuvant chemoradiation is probably better than surgical resection alone for patients with potentially curable disease, but only validation of this approach by CALGB-9781 can justify this as a new "proven" standard-of-care in the United States. 5. A pathologic complete response to neoadjuvant therapy is the strongest predictor of long-term survival. 6. 5-FU, by either short course or protracted continuous infusion, comprises the backbone of combination chemotherapy in combined-modality design. 7. Radiation therapy should be given at standard 1.8 to 2 Gy/fraction without a scheduled break. 8. Only by enrolling sufficient numbers of patients in prospective clinical trials will clinicians be able to further define the optimal sequencing and actual necessity of each individual component of combined-modality therapy. PMID- 9347834 TI - Barrett's esophagus. AB - The columnar replacement of squamous epithelium in the lower esophagus is the result of gastroesophageal reflux. Whether the squamous cells are replaced or undergo metaplasia is still conjectural. This neoepithelium is unstable in the presence of continued reflux and prone to complications of stricture, ulceration, and adenocarcinoma. Considerable evidence supports the hypothesis that duodenal contents play a role in the development of Barrett's esophagus and its complications. The increasing incidence of adenocarcinoma in Barrett's esophagus is of concern in the Western World. Surveillance programs in some centers have been successful in early diagnosis, and excellent survival periods have been reported following resection in these cases. Both medical and surgical antireflux treatment is successful in symptom relief, but even in the absence of symptoms, reflux may continue. Surgery offers better overall results than proton pump inhibition of gastric acid and has been more popular since less aggressive (minimally invasive) techniques have been popularized. Mucosal ablation and antireflux measures by medicine or surgery are still in the experimental stages but hold considerable promise for the future. PMID- 9347836 TI - Esophagectomy for cancer. AB - We are faced with the challenge of treating, palliating, or curing esophageal cancer, considered by some to be the most insidious of cancers. This article acquaints the reader with historic milestones regarding resection of esophageal cancer, preoperative evaluation and preparation, and the various techniques currently being used to perform esophagectomy for cancer. PMID- 9347837 TI - Endoscopic therapy of esophageal cancer. AB - In the management of esophageal cancer, endoscopy has evolved from a tool used to provide biopsy confirmation of suspected tumor to an integral part of the staging and ongoing treatment of patients. Endoscopic ultrasound is currently the most accurate means for T and N staging. Improved endoscopic techniques like dye staining and aggressive biopsy protocols can identify very early stage tumors in high-risk groups and allow curative surgery. Patients with early-stage tumors who are not surgical candidates can also be treated with endoscopic mucosectomy, photodynamic therapy, or Nd:YAG laser and still have a chance of long-term cure. Palliation of advanced tumors remains the major role of endoscopy in patients with esophageal cancer. A variety of techniques have proven effective over the years, including dilatation, laser, and rigid prostheses. Newer developments like bipolar probes, injection therapy, photodynamic therapy, and brachytherapy offer potential applications. The development and continuing improvements in both coated and uncoated expandable metal stents have been perhaps the greatest recent advance in endoscopic palliation of malignant dysphagia and esophagorespiratory fistulas. With the increasing array of endoscopic treatments and palliative techniques, emphasis must be placed on considering functional status; tumor characteristics like stage, location, and shape; patient wishes; and local expertise in tailoring treatment plans for each situation. PMID- 9347838 TI - Random musings on "why". PMID- 9347839 TI - Management of blunt splenic trauma: significant differences between adults and children. AB - BACKGROUND: Although highly successful in children, nonoperative management of blunt splenic injury in adults is less defined. The purpose of this study was to determine whether mechanism of injury, grade of splenic injury, associated injuries, and pattern of injury differ between adults and children (younger than 15 years of age). METHODS: Four hundred eleven patients (293 adults and 118 pediatric patients) with blunt splenic injury were admitted to an affiliated adult/pediatric trauma program from 1989 to 1994. Computed tomography (CT) scans were interpreted in a blinded fashion. Mechanism of injury was significantly different for adults versus children (p < 0.05): motor vehicle crash (66.9% versus 23.7%), motorcycle (8.8% versus 0.8%), sports (2.4% versus 16.9%), falls (8.8% versus 25.4%), pedestrian/automobile (4.4% versus 11.0%), bicycle (1.4% versus 9.3%), and other (7.3% versus 12.7%). RESULTS: Higher injury severity scores, lower Glasgow Coma Scales, and higher mortality indicated that the adults were more severely injured than the children. Fifty-nine percent of the adults and 7% of the children required immediate laparotomy for splenic injury. Both CT grade and quantity of blood on CT predicted the need for exploration in adults but not in children. An injury severity score above 15 and high-energy mechanisms correlated with the need for operative intervention. CONCLUSIONS: Rather than children simply being physically different, they are injured differently than adults, hence the high rate of nonoperative management. PMID- 9347840 TI - Prospective study of the features, indications, and surgical treatment in 513 consecutive patients affected by Crohn's disease. AB - BACKGROUND: The aim of this prospective study was to elucidate the features, indications, and surgical treatment in patients affected by complications of Crohn's disease. METHODS: Between January 1985 and July 1996, 513 consecutive patients (248 male, 265 female; mean age, 38 years) were operated on for 542 occurrences of Crohn's disease. Data were collected prospectively. RESULTS: Indications for abdominal surgery were often multiple but included failure of medical management (n = 220), obstruction (n = 94), intestinal fistula (n = 68), mass (n = 56), abdominal abscess (n = 33), hemorrhage (n = 7), and peritonitis (n = 9). Four hundred sixty-four abdominal procedures were performed, necessitating 425 intestinal resections and 97 stricture plasties. The use of stricture plasty was more common in the second half of the study (16.0% versus 7.3%, second half versus first half; p < 0.01). Perioperative complications occurred in 75 of the 464 abdominal operations (16%). There were no deaths. One hundred thirty patients (25%) required operation for perineal complications of Crohn's disease. The presence of Crohn's disease in the rectal mucosa was associated with a higher risk for permanent stomas in patients requiring operation for treatment of perianal Crohn's disease (67% versus 11%; p < 0.001). CONCLUSIONS: Patterns of surgical treatment in Crohn's disease are changing, with more emphasis on nonresectional options. The presence of rectal involvement significantly increases the need for a permanent stoma in patients with perianal Crohn's disease. PMID- 9347841 TI - Mammographic density in women on postmenopausal hormone replacement therapy. AB - BACKGROUND: Studies have suggested that mammographic density and pattern are affected by hormone replacement therapy (HRT) and may influence breast diagnosis. Because 40% of breast cancers diagnosed at our center are mammographically detected while still clinically occult, mammographic sensitivity is crucial. For this reason we studied the effect of HRT on mammographic density. METHODS: During a period of 18 months we studied consecutive women older than 54 years attending for breast screening. We recorded HRT use and dosing regimes. A breast density score (BDS) was developed and applied to all mammograms. RESULTS: Mammograms of 148 HRT users were compared with those of 158 nonusers. HRT users had a significantly higher mean density score (4.7 versus 3.4; p < 0.001). Only 11% of non-HRT users had high scores compared with 37% among HRT users (p < 0.001). The significant difference remained when women were stratified by age. Duration of HRT (longer or shorter than 5 years) did not affect density scores. CONCLUSIONS: HRT is associated with a significant increase in breast density. In turn, density and mammographic sensitivity are related. The possibility that increased breast density will hamper mammographic diagnosis of clinically occult cancers is worrisome. PMID- 9347842 TI - Risk analysis of coronary bypass surgery after acute myocardial infarction. AB - BACKGROUND: Current strategies for management of acute myocardial infarction (MI) include thrombolysis, angioplasty, and coronary bypass surgery singly or in combination. This study was designed to identify contemporary risk factors for coronary bypass surgery among patients in this high-risk group. METHODS: Between June 1992 and December 1995, 1181 consecutive patients underwent isolated coronary bypass surgery. Of these, 316 underwent coronary bypass surgery within 21 days of MI. Mean age was 65 years (range, 33 to 87 years), and 73% were male. There were 166 patients with stable angina (group 1), 107 patients with unstable angina requiring intravenous nitroglycerin for a control of ischemia (group 2), 20 patients with angina requiring intraaortic balloon counterpulsation for stabilization (group 3), and 23 patients with severe postinfarction ischemia complicated by cardiogenic shock (group 4). RESULTS: The overall in-hospital mortality rate was 5.1% (16 of 316), which was higher (p < 0.05) than the 2.5% (22 of 865) among patients undergoing coronary bypass surgery without recent myocardial infarction. Mortality increased with severity of clinical preoperative status and was 1.2% in group 1, 3.7% in group 2, 20.0% in group 3, and 26% in group 4. Serious postoperative morbidity occurred in 7.3% of patients. Multivariate logistic regression analysis identified preoperative intraaortic balloon counterpulsation, left ventricular dysfunction, and renal insufficiency as the only independent correlates of mortality. CONCLUSIONS: Coronary bypass surgery can be safely performed in stable patients at any time after acute MI, with an operative mortality similar to elective surgery. Thus, in this era of medical cost containment, there is no apparent indication for prolonged stabilization attempts that delay surgery. PMID- 9347843 TI - Laparoscopic-assisted ileocolic resections in patients with Crohn's disease: are abscesses, phlegmons, or recurrent disease contraindications? AB - BACKGROUND: Because of the inflammatory nature of Crohn's disease, ileocolic resections are often difficult to perform, especially if an abscess, phlegmon, or recurrent disease at a previous ileocolic anastomosis is present. Our goal was to determine whether the above factors are contraindications to a successful laparoscopic-assisted ileocolic resection. METHODS: Between 1992 and 1996, 46 laparoscopic-assisted ileocolic resections were attempted. Fourteen patients had an abscess or phlegmon treated with bowel rest before operation (group I), 10 patients had recurrent Crohn's disease at the previous ileocolic anastomosis (group II), and 22 patients had no previous operation and no phlegmon or abscess associated with their disease (group III). These groups were compared with each other and with 70 consecutive open ileocolic resections for Crohn's disease during the same time period (group IV). RESULTS: Operative blood loss and time were greater in group IV than in groups I, II, and III (245 versus 151, 131, and 195 ml, respectively, and 202 versus 152, 144, and 139 minutes, respectively). Conversion to open procedure occurred in 5 patients (group I, 1 [7%]; group II, 2 [20%]; group III, 2 [9%]). Morbidity was highest in group IV (21% versus 0%, 10%, and 10%, respectively). Only one patient died (group IV, 1%). Length of hospital stay was longest in group IV (7.9 versus 4.8, 3.9, and 4.5 days, respectively). CONCLUSIONS: The laparoscopic-assisted approach to Crohn's disease is feasible and safe with good outcomes. Co-morbid preoperative findings such as abscess, phlegmon, or recurrent disease at the previous ileocolic anastomosis are not contraindications to a successful laparoscopic-assisted ileocolic resection in select patients. PMID- 9347844 TI - Delayed gastric emptying affects outcome of Nissen fundoplication in neurologically impaired children. AB - BACKGROUND: Nissen fundoplication (NF) has a relatively high failure rate in neurologically impaired children with gastroesophageal reflux (GER). In 1990 we began to use routine technetium 99m sulfur colloid emptying scans and pyloroplasty with NF for delayed gastric emptying (DGE) in our neurologically impaired patients. The aim of this study was to determine the influence of DGE and pyloroplasty on the outcome of NF in neurologically impaired children. METHODS: One hundred neurologically impaired children underwent NF by a single surgeon between August 1986 and July 1995. Beginning in January 1990 emptying scans were routinely obtained, and patients with DGE underwent pyloroplasty with NF. Outcome analysis was performed for recurrence/wrap failure and other parameters. Mean follow-up was 5.8 years, with a minimum of 18 months. RESULTS: DGE was found in 35 (65%) of the 54 children who had emptying scans. All 11 children with normal scans had successful NF without recurrent reflux (100%). Forty (93%) of 43 children who underwent pyloroplasty and NF had successful outcomes. Thirty-eight children underwent NF without evaluation of gastric emptying with success in 30 of them (78.9%). Overall success improved from 34 (83%) of 41 in the first half of the study, when 3 (7%) of 41 children underwent emptying scans, to 55 (93%) of 59 in the second half, when 51 (86%) of 59 of the children underwent emptying scans. CONCLUSIONS: DGE is common in neurologically impaired children with GER. NF in children with normal gastric emptying has a high probability of success. Pyloroplasty improves the outcome of NF in children with DGE. Neurologically impaired children should be evaluated for DGE before operation for GER. PMID- 9347846 TI - Incidence and therapeutic implications of synchronous colonic pathology in colorectal adenocarcinoma. AB - BACKGROUND: The presence of synchronous benign and malignant colonic pathology may influence the magnitude of surgery for colorectal adenocarcinoma. The aim of this prospective study was to quantitate the need for a more extensive surgical procedure because of synchronous pathology in colorectal cancer patients. METHODS: Between 1984 and 1996, 235 consecutive patients were treated for colorectal adenocarcinoma. Preoperative survey of the colon in 228 patients included colonoscopy (91%) and double contrast barium enema (35.7%). Seven patients were excluded for incomplete preoperative survey because of perforating or obstructing colon carcinoma or acute ulcerative colitis. RESULTS: One hundred four patients (45.6%) had the following synchronous colonic lesions: benign polyps (68 patients, 29.8%), diverticular disease (30, 13.1%), ulcerative colitis (10, 4.4%), synchronous adenocarcinoma (8, 3.5%), and Crohn's colitis (3, 1.3%). Pathologic examination demonstrated three additional synchronous adenocarcinomas for a total of 11 patients (4.9%). Twenty-five (11%) required more extensive surgery than dictated by the primary cancer. Of these 25 patients, 17 had a benign or premalignant condition associated with their carcinoma and 8 had a synchronous carcinoma. Seventeen patients underwent a sphincter-saving procedure. Of the remaining eight patients requiring sphincter ablation, seven were needed because of a synchronous nonmalignant lesion, rather than because of the primary tumor. CONCLUSIONS: In our patient population, the incidence of synchronous colorectal lesions was 45.6%. Synchronous colorectal cancer occurred in 4.9%. In 11%, the presence of synchronous colorectal lesions made the surgical procedure more extensive than that dictated by the primary cancer, and in 3%, the need for a sphincter ablating procedure was dictated by a synchronous nonmalignant lesion. PMID- 9347845 TI - Implementation of a clinical pathway decreases length of stay and hospital charges for patients undergoing total colectomy and ileal pouch/anal anastomosis. AB - BACKGROUND: Clinical pathways are increasingly being used by hospitals to improve efficiency in the care of certain patient populations; however, little prospective data are available to support their use. This study examined whether using a clinical pathway for patients undergoing ileal pouch/anal anastomosis, a complex procedure in which we had extensive practical experience, affected hospital charges or length of stay (LOS). METHODS: A clinical pathway was developed to serve patients undergoing elective total colectomy and ileal pouch/anal anastomosis. All operations were performed by two attending physicians (J.E.F., M.S.N.). Before implementation, 10 pilot patients were prospectively monitored to ensure that hospital charges were accurately generated. In addition, charge audits were performed by an outside agency to verify the accuracy of the hospital bills. The pathway was then implemented, and 14 patients were prospectively analyzed. RESULTS: In all patients the principal diagnosis was ulcerative colitis, with the exception of three patients with familial polyposis. Mean external audit charges were within 2% of the hospital bills; therefore the hospital bills were used in all calculations. The mean LOS decreased from 10.3 days to 7.5 days (p = 0.046) for patients on the pathway versus pilot patients. Mean hospital charges also decreased significantly, from $21,650 to $17,958 per patient (p = 0.005). CONCLUSIONS: Implementation of a clinical pathway, even for an operation in which the surgeon has much experience, is an effective method for reducing LOS and charges for patients. This is likely the result of interdisciplinary cooperation, elimination of unnecessary interventions, and streamlined involvement of ancillary services. These results support the development of clinical pathways for procedures that involve routine preoperative and postoperative care. In addition, the benefits of clinical pathways should increase proportionally with increasing case volume for a particular procedure. PMID- 9347847 TI - Measuring postoperative complications in general surgery patients using an outcomes-based strategy: comparison with complications presented at morbidity and mortality rounds. AB - BACKGROUND: This study was undertaken to compare the incidence of adverse postoperative outcomes recorded in a prospective general surgery database with that identified through weekly morbidity and mortality (M&M) rounds and to measure the impact of feedback of information to the providers of care. METHODS: Data were collected on patients admitted to one general surgery service between October 1, 1995, and May 15, 1996, and recorded in a computer database. Postoperative complications were graded in severity from I (minor) to IV (mortality). RESULTS: Of 479 admissions entered into the database during the study period, 325 (311 patients) led to operations and were further analyzed. Admissions resulting in complications were associated with longer hospital stays, regardless of complication grade, compared to uncomplicated admissions (p < 0.01). A total of 29 of 106 patients with postoperative complications were presented at M&Ms (27.4%). Whereas 15.4% of database patients with grade I complications were presented at M&Ms, this proportion increased to 22.2% for grade IIa, 34.8% for grade IIb, 33.3% for grade III, and 87.5% for grade IV. (p < 0.05 for grade I, IIa, and IIb compared to grade IV). A total of 58 of 142 patients in the first part of the study period developed complications (40.8%), compared to 53 of 183 patients in the second part of the study (29%, p = 0.034). CONCLUSIONS: Although most severe complications are recorded at M&M rounds, a large proportion of complications remain unreported. Monitoring of outcomes may contribute to improvements in quality of care. PMID- 9347849 TI - Abdominal operations in patients with cirrhosis: still a major surgical challenge. AB - BACKGROUND: Hepatic transplantation and portasystemic shunts can be safely performed in patients with advanced liver disease, whereas other abdominal procedures appear to have a much higher mortality rate. This study reviews the outcomes of patients with cirrhosis after the full spectrum of abdominal operations. METHODS: In a 12-year period, 92 patients diagnosed with cirrhosis required either an emergent or elective abdominal operation. There were four categories of operations: cholecystectomy in 17 patients, hernia in 9, gastrointestinal tract in 54, and other procedures in 12. Fifty-five clinical, laboratory, and operative variables were analyzed to identify factors predictive of poor outcome. RESULTS: Coagulopathy developed in 24 patients (27%) and sepsis in 15 (16%). The mortality rate after emergent operations was 50%, compared to 18% for elective cases (p = 0.001). Other factors that predicted mortality included the presence of ascites (p = 0.006), encephalopathy (p = 0.002), and elevated prothrombin time (p = 0.021). The mortality in Child's class A patients was 10%, compared to 30% in class B and 82% in class C patients. CONCLUSIONS: Patients with cirrhosis undergoing elective or emergent operations are at a significant risk of developing postoperative complications leading to death. The most accurate predictor of outcome is the patient's preoperative Child's class. PMID- 9347848 TI - Epidermal growth factor and human growth hormone accelerate adaptation after massive enterectomy in an additive, nutrient-dependent, and site-specific fashion. AB - BACKGROUND: After massive enterectomy (ME), remnant intestine undergoes compensatory adaptation. Epidermal growth factor (EGF) and human growth hormone (hGH) have each been shown to enhance total length small intestine nutrient transport after ME. This study aims to determine the differential effects of EGF and hGH on proximal and distal small intestinal remnants after ME. METHODS: New Zealand white rabbits underwent 70% mid-jejunoileal resection. After 1 week, animals received hGH (0.2 mg/kg/day), EGF (1.5 micrograms/kg/hr), hGH + EGF, or vehicle (equal volume) for 7 days. Sodium-dependent uptake of glucose, glutamine, alanine, leucine, and arginine into brush border membrane vesicles was quantitated. Serum insulin-like growth factor-I concentrations as well as proximal and distal villus and microvillus heights were measured. IGF binding protein-3 and -4 mRNA expression was determined in full-thickness proximal and distal gut remnants. RESULTS: Concomitant hGH and EGF treatment up-regulates glucose (100%), glutamine (80%), and leucine (60%) transport in the proximal remnant; alanine (150%) and arginine (400%) transport in the distal remnant; and microvillus height (25% to 35%) both proximally and distally. Serum IGF-I levels and gross villus heights were not different among groups. CONCLUSIONS: Co infusion of hGH and EGF accelerates intestinal adaptation after ME in an additive, nutrient-dependent, and site-specific fashion via enhanced nutrient transport as well as microvillus hypertrophy. PMID- 9347850 TI - Use of the rapid/shallow breathing index as an indicator of patient work of breathing during pressure support ventilation. AB - BACKGROUND: Measuring patient work of breathing (WOBpt) has been suggested to provide safe, aggressive weaning from mechanical ventilation. We compared WOBpt and pressure-time-product (PTP) to routine weaning parameters [breath rate (f), tidal volume (VT), frequency/tidal volume ratio (f/VT)] at different levels of pressure support ventilation (PSV). METHODS: Fifteen patients in the surgical intensive care unit requiring prolonged weaning (more than 3 days) were entered in the study. A balloon-tipped esophageal catheter was placed and position confirmed by inspection of pressure and flow waveforms. Each patient was randomly assigned to breathe with 5, 10, 15, and 20 cm H2O of PSV. After 30 minutes, 40 breaths were recorded and analyzed. Measurement of WOBpt PTP, f, VT, and f/VT were made using the Bicore CP-100 monitor. Mean values for each parameter were calculated. PTP and WOBpt were plotted against f/VT to determine correlation coefficient. RESULTS: PTP, WOBpt and f/VT decreased in a stepwise fashion as PSV was increased. The f/VT correlated most closely with WOBpt (r = 0.983) and PTP (r = 0.972). Monitoring f alone also correlated with WOBpt (r = 0.894) and PTP (r = 0.881). All patients were weaned from the ventilator (mean duration, 22 +/- 5.9 days). Nine patients required tracheostomy before final liberation from the ventilator (mean duration, 22 +/- 5.9 days). Nine patients required tracheostomy before final liberation from the ventilator. CONCLUSIONS: Direct measurement of WOBpt is invasive, expensive, and' may be confusing to clinicians. Monitoring f/VT may be useful when changing PSV during weaning. PMID- 9347851 TI - Perivascular and intralesional tissue necrosis after hepatic cryoablation: results in a porcine model. AB - BACKGROUND: Cryosurgical ablation of malignant hepatic tumors is being increasingly used for definitive treatment of metastatic colorectal and primary hepatic tumors. The lack of tumor necrosis near vessels that results from inadequate freezing may contribute to local recurrence and thus limit the applications of this therapy. This study was designed to determine whether single freeze cryoablation could cause necrosis of both the pervascular and intralesional hepatic parenchyma. METHODS: Ten pigs were treated with one 15 minute cycle of cryoablation. Five additional animals were treated with overlapping cryolesions to simulate a double freeze. After 24 hours, animals underwent reoperation with portal vein cannulation and infusion of formalin. Serial sectioning and hematoxylin and eosin staining of cryolesions were performed. RESULTS: Complete cell death was visualized within all cryolesions. There was no difference between once or twice-frozen tissue. Vessels within or adjacent to cryolesions showed necrosis of hepatic tissue up to the vessel wall. No sections revealed incomplete necrosis of perivascular hepatic parenchyma. CONCLUSIONS: Single-freeze cryoablation results in necrosis of intralesional hepatic parenchyma without added benefit from repeat freezing. Complete necrosis of the perivascular tissue suggests that cryosurgical ablation can effectively cause necrosis immediately adjacent to vessels without concerns of incomplete ablation resulting from the heat sink effect. PMID- 9347852 TI - Carcinoma involving the gallbladder in elderly patients presenting with acute cholecystitis. AB - BACKGROUND: The unexpected intraoperative finding of a cancerous gallbladder has become particularly problematic, because cancer recurs rapidly after laparoscopic cholecystectomy. It would be desirable to identify the patients of greatest risk for gallbladder cancer before operation. After several elderly patients presenting with acute cholecystitis were found to have gallbladder cancer, we performed the following study. METHODS: Records of patients (60 years of age or older, 1987 to 1995) with an admitting diagnosis of acute cholecystitis and symptoms including right upper quadrant pain, nausea, vomiting, fever, and leukocytosis were reviewed. RESULTS: Eighty patients were included in the study. Carcinoma involving the gallbladder was found in seven patients; six had primary and one had metastatic carcinoma. The 73 patients without cancer underwent cholecystectomy. The differences between the noncancer and cancer patients included age (68 +/- 7 versus 74 +/- 8 years, p < 0.05), total bilirubin (mg/dl, 1.5 +/- 1.5 versus 3.7 +/- 3.4, p < 0.01), alkaline phosphatase (IU/L, 179 +/- 132 versus 369 +/- 226, p < 0.01), and aspartate aminotransferase (IU/L, 77 +/- 93 versus 158 +/- 157, p < 0.05). CONCLUSIONS: Additional work-up and open cholecystectomy should be considered in elderly patients presenting with apparent acute cholecystitis, especially when liver functions are abnormal. PMID- 9347853 TI - Activated inflammatory cells are associated with plaque rupture in carotid artery stenosis. AB - BACKGROUND: Histologic studies of carotid plaques have demonstrated an association between symptomatic disease and plaque rupture. The purpose of this study was to characterize the cellular changes associated with plaque rupture. METHODS: Carotid plaques were obtained from 61 patients undergoing carotid endarterectomy for established indications. Plaques were fixed in formalin, embedded in paraffin, and stained with hematoxylineosin and Movat pentachrome stain to demonstrate plaque structure. Each plaque was examined with light microscopy and classified as containing evidence of plaque rupture (n = 29) or no plaque rupture (n = 32). By using immunohistochemical staining, the fibrous cap was examined for the presence of smooth muscle cells (alpha-actin), macrophages (KP-1), T lymphocytes (UCHL-1), and cell activation (HLA-DR). Data were analyzed with chi-squared analysis. RESULTS: With plaque rupture, macrophages and T lymphocytes were significantly more common than in specimens without evidence of rupture. Similarly, macrophages and T lymphocytes expressing HLA-DR were more often found in sections containing plaque rupture. Furthermore, vascular smooth muscle cells were more common with intact fibrous caps and were diminished with cap thinning. CONCLUSIONS: Rupture of the fibrous cap in carotid artery lesions is associated with increased numbers of macrophages and T lymphocytes, which are in an activated state. The activated inflammatory cells may release cytokines or metalloproteinases, which may be responsible for loss of the fibrous cap. Thus inflammation appears to play a role in the pathogenesis of the neurologic symptoms associated with carotid artery stenosis. PMID- 9347855 TI - Long-term follow-up of auxiliary orthotopic liver transplantation for the treatment of fulminant hepatic failure. AB - BACKGROUND: Auxiliary orthotopic liver transplantation (AOLT) was investigated as a bridge to native liver recovery in patients with fulminant hepatic failure (FHF). METHODS: In the last 5 years seven patients with FHF were treated with AOLT at our institution. Five patients underwent resection of the native left lobe and orthotopic replacement with a donor left lobe (n = 3) or left lateral segment (n = 2). Two patients underwent left trisegmentectomy and whole liver auxiliary grafting. Conventional immunosuppression was used in all patients. RESULTS: One patient had poor initial graft function and required retransplantation. Native liver function returned to normal in the six other patients. Immunosuppression was gradually tapered and completely discontinued in three patients, allowing for atrophy of the allograft. The allograft was removed in the other four patients. Despite evidence of native liver regeneration, two patients with aplastic anemia died after allograft removal. Four patients are alive at a mean follow-up of 3.5 years. CONCLUSIONS: AOLT is technically feasible, rapidly restores liver function, and should be considered an important alternative to standard orthotopic liver transplantation (OLT) in the treatment of FHF. AOLT has the advantage that patients transplanted for FHF are not committed to lifelong immunosuppression with its attendant risks. PMID- 9347854 TI - A newly identified pattern of K-ras mutations at codons 12 and 13 is associated with long-term survival in colorectal cancer. AB - BACKGROUND: Although K-ras mutations reportedly occur in 40% to 60% of all colorectal carcinomas, the relationship between specific mutations and clinical outcome is unclear. The purpose of this study was to assess the frequency and types of K-ras mutations in 89 colorectal cancer patients, comparing groups with short-term (less than 5 years) and long-term (more than 10 years) survival. METHODS: The group was divided into four cohorts by survival and modified Dukes classification (Dukes B2 and C2). DNA was extracted from formalin-fixed paraffin embedded archival material. Mutational status was analyzed using a modification of allele-specific-polymerase chain reaction. RESULTS: Mutations in codon 12 were found in 11.2% of tumors, and 83% of tumors had mutations in codon 13. Gly > Asp accounted for 85.2% of the mutations. Tumors with mutations in both codon 12 and codon 13 occurred significantly more frequently in the long-term (21.3%) versus the short-term (2.4%) survival group. Gly > Asp mutations in either codon were related to long-term survival, and 80% of long-term survivors with mutations in both codons had Gly > Asp mutations in both. CONCLUSIONS: Simultaneous mutation in codons 12 and 13 of the K-ras gene appears to be a positive prognostic indicator in colorectal cancer. PMID- 9347856 TI - Chances of cure are not compromised with sphincter-saving procedures for cancer of the lower third of the rectum. AB - BACKGROUND: The goal of this study was to compare patterns of recurrence and long term outcome after sphincter-saving procedures (SSPs) and abdominoperineal resection (APR) in patients with tumors located in the lower third of the rectum. METHODS: We reviewed the charts of 1001 patients operated on for primary rectal adenocarcinoma between 1980 and 1991. All patients with tumors located between 5 and 7 cm from the anal verge and treated with curative intent were included. RESULTS: Of the 261 patients who met our criteria, 162 had undergone SSP and 99 had undergone APR. The local recurrence rates for SSP and APR were 8% and 11%, respectively (p = 0.41), and the distant metastases rates were 23% and 28%, respectively (p = 0.35). Recurrence and distant metastases rates for SSP and APR, respectively, did not differ by TNM classification: state I, 10% versus 9% (p = 0.9); stage II, 25% versus 43% (p = 0.13); and stage III, 56% versus 57% (p = 0.92). Five-year disease-free survival rates for SSP and APR patients were 70.5% and 62.3%, respectively (p = 0.2). CONCLUSIONS: Tumors in the lower third of the rectum can be treated with sphincter-saving procedures without compromising the chance of cure. PMID- 9347857 TI - Mucin-hypersecreting intraductal neoplasms of the pancreas: a precursor to cystic pancreatic malignancies. AB - BACKGROUND: Muncin-hypersecreting intraductal pancreatic neoplasms were first described in 1982 and have been observed in increasing numbers since. They are observed primarily by endoscopic retrograde cholangiopancreatography (ERCP) and are characterized by an intraductal papillary neoplasm that secretes thick mucin, causing pancreatic duct dilatation and obstructive pancreatitis. METHODS: Twenty patients are presented, 14 male and six female, with an average age of 59 +/- 11 years. All patients presented with abdominal pain, and most had nausea and vomiting, weight loss, and documented pancreatitis. Of the preoperative studies, ERCP was positive in all patients. Computed tomography scan, endoscopic ultrasonogram, and cytologic findings were less sensitive. Tumor markers were only positive in one patient. All 20 patients were treated surgically. Nine underwent Whipple procedure, one patient had a total pancreatectomy, and nine had distal pancreatic resections. The first patient in the series did not have a pancreatic resection, and his disease evolved into a lethal cystadenocarcinoma causing his death 99 months later. RESULTS: Histopathologic findings were interpreted as borderline malignant in 17 of the 20 patients, and three patients had evidence of invasive adenocarcinoma. Two of these three patients had nodal or distant metastases at the time of diagnosis, and all three died of adenocarcinoma. Seventeen of the patients are alive and well, although two of three with positive pancreatic margins have had recurrent symptoms and have been successfully reresected. CONCLUSIONS: The mucin-producing intraductal papillary tumor of the pancreas is a newly described variant of pancreatic cancer. It presents with symptoms of pancreatitis and has a progressive but more indolent course than the more lethal invasive ductal cancers. Patients with unexplained pancreatitis should undergo ERCP investigation, and aggressive surgical therapy should be carried out because the prognosis for this lesion, when appropriately treated, is more favorable than the usual pancreatic cancer. PMID- 9347858 TI - Transjugular intrahepatic portosystemic shunt versus H-graft portacaval shunt in the management of bleeding varices: a cost-benefit analysis. AB - BACKGROUND: Transjugular intrahepatic portosystemic shunt (TIPS) is popular in treating portal hypertension because of its perceived efficacy and cost benefits, although it has never been compared with surgical shunting in a cost-benefit analysis. This study was undertaken to determine the cost benefit of TIPS versus small-diameter prosthetic H-graft portacaval shunt (HGPCS). METHODS: Cost of care was determined in 80 patients prospectively randomized to receive TIPS or HGPCS as definitive treatment for bleeding varices, beginning with shunt placement and including subsequent admissions for complications or follow-up related to shunting. RESULTS: Patients were similar in age, gender, severity of illness/liver dysfunction, and urgency of shunting. After TIPS or HGPCS, variceal rehemorrhage (8 versus O, respectively; p = 0.03), shunt occlusion (13 versus 4; p = 0.03), shunt revision (16 versus 4; p < 0.005), and shunt failure (18 versus 10; p = 0.10) were compared; all were more common after TIPS. Through the index admission, TIPS cost $48,188 +/- $43,355 whereas HGPCS cost $61,552 +/- $47,615. With follow-up, TIPS cost $69,276 +/- $52,712 and HGPCS cost $66,034 +/- $49,118. CONCLUSIONS: Early cost of TIPS was less than, though not different from, cost of HGPCS. With follow-up, costs after TIPS mounted. The initially lower cost of TIPS is offset by higher rates of subsequent occlusion and rehemorrhage. PMID- 9347859 TI - Bronchial carcinoid--twenty years' experience defines a selective surgical approach. AB - BACKGROUND: The purpose of this study was to look at the clinical behavior of bronchial carcinoids and clarify a surgical approach. METHODS: Eighty-four patients resected for bronchial carcinoids were retrospectively reviewed for clinicopathologic variables, surgical management, and outcome. Tumors were considered "typical" or "atypical" based on histologic features. "Conservative" surgery signified lung parenchyma-sparing procedures. Survival analysis was performed using standard statistical methods. RESULTS: Most patients presented with an abnormal routine chest x-ray. One patient had the carcinoid syndrome. Computed tomography scan reliably predicted lymph node status and bronchoscopic biopsy diagnosed carcinoids with 70% success. Fifteen "conservative" procedures were performed. Fifteen percent of patients had atypical carcinoids, 12% presented with lymph node metastases, and 6 patients had tumorlets associated with the primary tumor. Overall survival rates were 93% and 82% at 5 and 10 years, respectively. Significantly decreased disease-free survival was found with atypical histology (p < 0.0001) and the presence of tumorlets (p = 0.02); lymph node involvement strongly tended toward poorer outcome. CONCLUSIONS: Bronchial carcinoids have a definite malignant potential predicted by atypical histology, presence of tumorlets, and lymph node involvement. These features can be identified with routine bronchoscopic biopsy, computed tomography scanning, and intraoperative assessment including frozen section. In the select group of patients without negative features, strong consideration should be given to performing a conservative resection. PMID- 9347860 TI - Crohn's disease and ulcerative colitis mucosal T cells are stimulated by intestinal epithelial cells: implications for immunosuppressive therapy. AB - BACKGROUND: Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory diseases, and their pathogenesis is attributed, in part, to alterations of the mucosal immune system. This study was designed to define the possible contribution of epithelial cells to the activation of lamina propria T lymphocytes (LPTs) in CD and UC. METHODS: LPTs isolated from CD, UC, and control surgical specimens were cocultured with freshly isolated allogeneic or autologous epithelial cells or epithelial cell lines. Resulting T-cell proliferation was evaluated by tritiated thymidine incorporation on day 5. RESULTS: When intestinal epithelial cells were used to stimulate mucosal T-cell proliferation, CD and UC LPTs were less responsive than control LPTs (p < 0.05 and p < 0.03, respectively). This difference between inflamed and control T cells was consistently observed by using a variety of different intestinal epithelial cell types. CONCLUSIONS: CD and UC mucosal T cells are hyporesponsive to activation by intestinal epithelial cells when compared with control LPTs. Elucidating the mechanism underlying the differential activation of CD and UC LPTs may help to better understand the immunopathogenesis of these conditions. PMID- 9347861 TI - An analysis of perioperative cholangiography in one thousand laparoscopic cholecystectomies. AB - BACKGROUND: We undertook this retrospective study to ascertain the proper role of perioperative cholangiography in the management of 1002 patients undergoing laparoscopic cholecystectomy for symptomatic cholelithiasis. METHODS: Nine hundred forty-one patients were categorized as being at high or low risk for choledocholithiasis according to the presence or absence of jaundice, pancreatitis, elevated bilirubin, alkaline phosphatase, serum glutamic oxaloacetic transaminase, or radiographic evidence of common bile duct stones (CBDSs). RESULTS: Intraoperative cholangiography (IOCG) and preoperative endoscopic retrograde cholangiopancreatography (ERCP) were equivalent in the detection of CBDSs, and laparoscopic common bile duct exploration (CBDE) was successful in 12 of the 21 patients (57%) in whom it was attempted. The ducts of the other 52 patients with CBDSs were successfully cleared by preoperative or postoperative ERCP. CONCLUSIONS: Laparoscopic IOCG is successful in detecting CBDS in high-risk patients and half of these ducts can be cleared laparoscopically. The incidence of CBDS in low-risk patients is 1.7%, a risk that does not warrant routine cholangiography. These data suggest ERCP should be reserved for those at-risk individuals in whom IOCG or laparoscopic duct clearance has been unsuccessful. PMID- 9347862 TI - Fine-needle aspiration of 697 palpable breast lesions with histopathologic correlation. AB - BACKGROUND: Fine-needle aspiration breast biopsy has been used increasingly as an alternative to excisional biopsy. The purpose of this study is to evaluate the accuracy of fine-needle aspiration with histopathologic confirmation. METHODS: A retrospective study was performed using a computer database over a 5-year period. All women who had had fine-needle aspiration breast biopsy with histopathologic confirmation of the diagnosis were included. Fine-needle aspirations were interpreted as malignant, suspicious, or benign. Histopathologic diagnosis included core-needle biopsy, open excisional biopsy, or mastectomy specimen. RESULTS: A total of 697 patients fulfilled the criteria. Only 5 (0.7%) of the specimens were inadequate for study. There were 401 total malignant fine-needle aspiration diagnoses, with only 3 false-positive specimens. All three were ductal hyperplasia, one from a previously radiated breast. There were 125 suspicious readings; 84 of these were malignant and 41 were false-suspicious specimens. Most of the false-suspicious lesions were fibrocystic disease. Of the 166 lesions interpreted as benign, there were 13 false-negative specimens. The test had a 97% sensitivity, 78% specificity, 92% positive predictive value, and 92% negative predictive value. CONCLUSIONS: Fine-needle aspiration is a sensitive test that can be useful as an adjunct in the diagnosis of breast cancer. "Malignant" and "benign" interpretations are highly predictive but must be used only in the context of other diagnostic modalities. "Suspicious" lesions require further investigation. PMID- 9347864 TI - Thoracoscopic esophagomyotomy for achalasia: maximum gain, minimal pain. AB - BACKGROUND: Achalasia can be effectively treated by either hydrostatic balloon dilatation or transthoracic modified Heller myotomy. The purpose of this study was to determine whether thoracoscopic methods could be used to achieve surgical results equal to the transthoracic approach with less pain. METHODS: Twenty-one patients (10 men, 11 women; median age 42 years) had the diagnosis of achalasia confirmed by manometry, radiography, and endoscopy. All had dysphagia; five had weight loss. Median duration of symptoms was 12 months (range: 1 to 360 months). Eleven patients had undergone previous unsuccessful hydrostatic dilatation. Mean esophageal diameter was 5.5 +/- 2.2 cm. RESULTS: All patients underwent attempted modified Heller myotomy through a left thoracoscopic approach. Three patients required conversion to thoracotomy. The myotomy was extended < 1 cm past the squamocolumnar junction. There was one intraoperative perforation and no postoperative complications. All patients were begun on a regular diet on the first postoperative morning. Median length of stay was 2 days, Median follow-up was 22 months (range: 1 to 52 months). Sixteen patients (80%) had excellent relief of their dysphagia. Two patients (10%) had good relief, and two patients had only a fair result, although even they claim to be much improved. CONCLUSIONS: Thoracoscopic Heller myotomy reproduces the superior results of open esophagomyotomy with a reduced hospitalization and reduced incisional pain and disability. PMID- 9347863 TI - Isolated upright gastroesophageal reflux is not a contraindication for antireflux surgery. AB - BACKGROUND: Patients with gastroesophageal reflux disease who reflux only in the upright position are thought to have a less severe abnormality. Controversy exists over whether these patients should be considered candidates for antireflux surgery. METHODS: A total of 224 consecutive patients with increased esophageal acid exposure on 24-hour pH monitoring were classified as having upright (n = 54), supine (n = 72), or bipositional (n = 98) reflux and were evaluated by manometry and endoscopy. Of these, 116 patients had a laparoscopic Nissen fundoplication. Their clinical outcome at a median of 12 months (range 4 to 44 months) was compared. RESULTS: Patients with upright reflux had a lower prevalence of a structurally defective lower esophageal sphincter, fewer hiatal hernias, and less esophageal injury when compared to those with bipositional reflux (p < 0.005). Excellent (asymptomatic) or good outcome (minor symptoms not requiring acid suppression therapy) was achieved in 86% of the patients with upright reflux, 90% of those with supine reflux, and 89% of those with bipositional reflux. CONCLUSIONS: Patients with upright reflux have less complicated, earlier disease and have results equivalent to those patients with supine and bipositional reflux after antireflux surgery. PMID- 9347865 TI - Liver transplantation in Ohio. AB - BACKGROUND: Since its inception in 1984, the Ohio Solid Organ Transplantation Consortium has tracked liver transplantation outcomes for its five member institutions. Presented herein is a 12-year summary of this data analyzed to determine whether, with increasing experience, outcomes have improved in a cost effective manner. METHODS: Between July 1984 and June 1996, 1,063 liver transplants were performed in Ohio in 943 patients (772 adults and 171 children), of which 943 were primary and 120 were retransplants (13%). Outcome comparisons were made for three eras: 1984-1988, 1988-1992, and 1992-1996. RESULTS: The percentage of urgent (United Network for Organ Sharing status 1 and 2) transplants has decreased (62% to 41%), whereas that of homebound patients has increased (38% to 59%). Average time on the waiting list has increased from 39 to 165 days, and the average length of stay has decreased from 44 to 27 days. Patient survival at 1-year increased in each era (64%, 80%, and 82%, respectively). Although actual hospital charges have remained relatively constant, they have decreased substantially when compared in 1985 dollars as corrected for inflation. CONCLUSIONS: Patients undergoing liver transplantation in Ohio are now listed earlier in the course of their disease and wait longer for their transplant, but enjoy a better chance of survival, have a shorter hospital stay, and a relatively less expensive operation. These data indicate that with increased experience, the Ohio Solid Organ Transplantation Consortium liver transplantation teams perform liver transplantation in a more cost-effective manner. PMID- 9347866 TI - Timing of carotid endarterectomy in patients with recent stroke. AB - BACKGROUND: There is little objective data to support the conventional wisdom of waiting 4 to 6 weeks after stroke to improve surgical outcome of subsequent carotid endarterectomy (CEA). We have aggressively pursued CEA in patients after recent stroke; in this study we report our results. METHODS: We performed 215 CEA procedures in 200 patients who presented with an indication of stroke within 6 months of CEA. Cervical block anesthesia was used 193 cases. The rest were performed with the patient under general anesthesia. RESULTS: Perioperative stroke rate was 1.4% (3/215), and operative mortality was 2% (4/200) (stroke mortality = 3.4%). There were four early occlusions. Shunts were used in 13.9%, patch closure in 8.4%, and eversion endarterectomy in 48% of cases. There was no correlation between timing of surgery, extent of infarct on computed tomography/magnetic resonance imaging, and postoperative neurologic complications with the occurrence of postoperative stroke (p = NS). During the same period, 1,922 patients underwent CEA for indications other than stroke, with a perioperative stroke rate and mortality rate of 1.1%. CONCLUSIONS: Selected patients presenting with a history of stroke and significant carotid artery disease can safely undergo early CEA with a mortality and morbidity comparable to patients undergoing CEA for other indications. PMID- 9347867 TI - The role of rectosigmoid neocolporrhaphy. AB - BACKGROUND: Vaginoplasty for congenital vaginal atresia, a component of the Mayer Rokitansky-Kuster syndrome, or for gender confirmation, may be achieved by several techniques. This report focuses on the efficacy of rectosigmoid neocolporrhaphy (RSNC) performed either primarily or secondarily after failure of another procedure. METHODS: Sixty patients underwent isoperistaltic RSNC, three primarily and 57 secondarily. The indication was vaginal atresia in 1 patient and gender dysphoria in 59 patients. RESULTS: All 60 patients survived and have a functional neovagina. One major complication, an anastomotic leak with colovaginal fistula, was treated by a temporary colostomy and later reconstruction through a combined anterior and posterior approach. Late complications were reversible stomal stenosis (six patients), reversible conduit narrowing (five patients), transient rhabdomyoblastosis (one patient), and temporary mucosal bleeding from hyperplasia (three patients). Thirty patients have regular intercourse, 12 patients have occasional intercourse, and the others feel "whole," with their intact desired sexual anatomy awaiting a suitable partner. CONCLUSIONS: The number of patients seeking vaginoplasty is increasing. Primary or secondary RSNC is a safe and effective method. PMID- 9347868 TI - The effect of early versus late fasciotomy in the management of extremity trauma. AB - BACKGROUND: Recent reports have demonstrated an increase in the number of complications associated with delayed timing of fasciotomy for trauma. This study examines the effectiveness of early (less than 12 hours) versus late (more than 12 hours) fasciotomy in the injured extremity. METHODS: This is a retrospective review of 88 patients undergoing fasciotomy for extremity trauma admitted to the University of Cincinnati from January 1990 through December 1995. Records were reviewed for demographics, compartment pressures, time and type of fasciotomy, complications, limb salvage, and mortality. Statistical analysis was determined with chi-squared, multivariant regression analysis, and Student's t test with significance at p less than 0.05. RESULTS: Sixty-one (69%) patients had fasciotomy performed before 12 hours and twenty-seven (31%) after 12 hours. Although the rates of infection differed significantly between the two groups (7.3% for early versus 28% for late), the rates of limb salvage and neurologic sequelae were similar. Age, mechanism, shock, associated injuries, and time to fasciotomy were not predictive of complications. CONCLUSIONS: Fasciotomy for trauma is most efficacious when performed early. However, when performed late, it results in similar rates of limb salvage as compared with early fasciotomy but at the increased risk of infection. These results support aggressive use of fasciotomy in extremity trauma regardless of time of diagnosis. PMID- 9347870 TI - Silicates, silicones and autoimmunity. PMID- 9347869 TI - beta 2-Glycoprotein I and antiphospholipid syndrome. PMID- 9347871 TI - Heparin-induced thrombocytopenia as an autoimmune disease--idiotypic evidence for the role of anti-heparin/PF4 autoantibodies. PMID- 9347872 TI - Melatonin--a possible link between sleep and the immune system. PMID- 9347873 TI - Anti Ro/La antibodies and their clinical association. PMID- 9347874 TI - Sex hormones and autoimmunity. PMID- 9347875 TI - Balneotherapy in autoimmune disease. AB - The mechanisms of action of balneotherapy in the treatment of autoimmune disease are not sufficiently clear. Although this therapy does not replace but rather complements conventional drug therapy, it is certainly beneficial in suitable cases. Additional controlled studies are needed to delineate the mechanisms of actions and the effectiveness of balneotherapy in autoimmune disease. PMID- 9347876 TI - Protooncogenes and autoimmunity. PMID- 9347877 TI - Antiperinuclear factor--a specific marker for rheumatoid arthritis. PMID- 9347878 TI - Nutrition and autoimmunity. PMID- 9347879 TI - Intracellular fluorescence polarization measurements by the Cellscan system: detection of cellular activity in autoimmune disorders. PMID- 9347880 TI - Copolymer 1 from the laboratory to FDA. PMID- 9347882 TI - On hepatitis C. PMID- 9347881 TI - Intracellular IL-1 alpha in fibroblasts as a possible endogenous mediator of joint damage in rheumatoid arthritis. PMID- 9347889 TI - Synergy, antagonism, and scientific process. PMID- 9347890 TI - Low-level radiation harmed humans near Three Mile Island. PMID- 9347892 TI - Environmental health gold mine in New Jersey. PMID- 9347893 TI - Growing pains in South America. PMID- 9347894 TI - A green light for new research. PMID- 9347895 TI - The estrogenic activity of phthalate esters in vitro. AB - A large number of phthalate esters were screened for estrogenic activity using a recombinant yeast screen. a selection of these was also tested for mitogenic effect on estrogen-responsive human breast cancer cells. A small number of the commercially available phthalates tested showed extremely weak estrogenic activity. The relative potencies of these descended in the order butyl benzyl phthalate (BBP) > dibutyl phthalate (DBP) > diisobutyl phthalate (DIBP) > diethyl phthalate (DEP) > diisiononyl phthalate (DINP). Potencies ranged from approximately 1 x 10(6) to 5 x 10(7) times less than 17beta-estradiol. The phthalates that were estrogenic in the yeast screen were also mitogenic on the human breast cancer cells. Di(2-ethylhexyl) phthalate (DEHP) showed no estrogenic activity in these in vitro assays. A number of metabolites were tested, including mono-butyl phthalate, mono-benzyl phthalate, mono-ethylhexyl phthalate, mon-n octyl phthalate; all were wound to be inactive. One of the phthalates, ditridecyl phthalate (DTDP), produced inconsistent results; one sample was weakly estrogenic, whereas another, obtained from a different source, was inactive. analysis by gel chromatography-mass spectometry showed that the preparation exhibiting estrogenic activity contained 0.5% of the ortho-isomer of bisphenol A. It is likely that the presence of this antioxidant in the phthalate standard was responsible for the generation of a dose-response curve--which was not observed with an alternative sample that had not been supplemented with o,p'-bisphenol A- in the yeast screen; hence, DTDP is probably not weakly estrogenic. The activities of simple mixtures of BBP, DBP, and 17beta-estradiol were assessed in the yeast screen. No synergism was observed, although the activities of the mixtures were approximately additive. In summary, a small number of phthalates are weakly estrogenic in vitro. No data has yet been published on whether these are also estrogenic in vitro. No data has yet been published on whether these are also estrogenic in vivo; this will require tests using different classes of vertebrates and different routes of exposure. PMID- 9347896 TI - Structure-activity relationships for xenobiotic transport substrates and inhibitory ligands of P-glycoprotein. AB - The multixenobiotic resistance phenotype is characterized by the reduced accumulation of xenobiotics by cells or organisms due to increased efflux of the compounds by P-glycoprotein (P-gp) or related transporters. An extensive xenobiotic database, consisting primarily of pesticides, was utilized in this study to identify molecular characteristics that render a xenobiotic susceptible to transport by or inhibition of P-gp. Transport substrates were differentiated by several molecular size/shape parameters, lipophilicity, and hydrogen bonding potential. Electrostatic features differentiated inhibitory ligands from compounds not catagorized as transport substrates and that did no interact with P gp. A two-tiered system was developed using the derived structure-activity relationships to identify P-gp transport substrates and inhibitory ligands. Prediction accuracy of the approach was 82%. We then validated the system using six additional pesticides of which tow were predicted to be P-gp inhibitors and four were predicted to be noninteractors, based upon the structure-activity analyses. Experimental determinations using cells transfected with the human MDR1 gene demonstrated that five of the six pesticides were properly catagorized by the structure-activity analyses (83% accuracy). Finally, structure-activity analyses revealed that among P-gp inhibitors, relative inhibitory potency can be predicted based upon the surface area or volume of the compound. These results demonstrate that P-gp transport substrates and inhibitory ligands can be distinguished using molecular characteristics. Molecular characteristics of transport substrates suggest that P-gp may function in the elimination of hydroxylated metabolites of xenobiotics. PMID- 9347897 TI - Lead exchange in teeth and bone--a pilot study using stable lead isotopes. AB - Stable lead isotopes and lead concentrations were measured in the enamel and dentine of permanent (n = 37) and deciduous teeth (n = 14) from 47 European immigrants to Australia to determine whether lead exchange occurs in teeth and how it relates to lead exchange in bone. Enamel exhibits no exchange of its European-origin lead with lead from the Australian environment. In contrast, dentine lead exchanges with Australian lead to the extent of approximately 1% per year. In one subject, trabecular bone from the tooth socket exchanged almost all its European lead with Australian lead over a a 15-year period (turnover of approximately 6% per year), similar to the approximately 8% per year proposed for lead turnover in trabecular bone. The repository characteristics of intact circumpulpal dentine were investigated by analyses of four sets of contiguous slices from six teeth: 1) a set consisting of slices with intact circumpulpal dentine and cementum; 2) a set in which these areas were removed; 3) another set consisting of slices with intact circumpulpal dentine and cementum; and 4) a set without cementum. These analyses show relatively small differences in isotopic composition between contiguous slices except that circumpulpal dentine appears to be the dominant control on lead concentration. There is a significant correlation (R2 = 0.19, p = 0.01, n = 34) of dentine lead concentration and rate of exchange with residence time from the country of origin and Australian lead, but there is no such correlation with enamel lead concentration. Analyses of permanent and deciduous teeth of subjects from other countries who have resided in Australia for varying lengths of time should resolve some of the questions arising from this pilot study. PMID- 9347898 TI - The metropolitan acid aerosol characterization study: results from the summer 1994 Washington, D.C. field study. AB - An ambient particle monitoring study was conducted in Washington, D.C. during the summer of 1994 as part of the Metropolitan Acid Aerosol Characterization Study (MAACS). Acid aerosol and inhalable (particulate matter with aerodynamic diameters < 10 micron; PM10) and fine (particulate matter with aerodynamic diameters < 2.5 micron; PM2.5) particle samples were collected for 24-hr periods (9 A.M.-9 A.M. EDT) on alternate days at six monitoring sites located throughout the greater Washington, D.C. area. Monitoring sites were located in both urban and rural areas and were generally situated along a southwest to northeast line due to the prevailing winds. Information on site characteristics, including population density and distance from the city center, was also obtained, as were data on meteorological parameters. Results from this study show strong correlations among the particulate measures, PM10, PM2.5, SO4(2-), and H+. These strong correlations resulted from the fact that PM2.5 comprised 77% of PM10, with SO4(2-)-related species accounting for 49% of total PM2.5. PM10, PM2.5, SO4(2-), and H+ concentrations were found to be uniform across the metropolitan Washington area. Spatial variation was found, however, for coarse particles (PM2.5-10) and NH3 concentrations. In our previous Philadelphia study, population density was an important determinant of spatial variation in coarse particles and NH3 concentrations, however, in Washington, D.C., population density was not associated with observed spatial patterns in coarse particle concentrations, but was an important determinant of NH3 concentrations. When data from one site (Reservoir) was excluded from the analysis, population density explained larger percentage of the variability in NH3 levels and became an important determinant of the H+/SO4(2-) ratio as well. Ambient H+ models developed from Philadelphia data were found to predict H+ concentrations in Washington, D.C. reasonably well, representing an improvement over measurements made at a single stationary ambient monitoring site. PMID- 9347899 TI - Airborne arsenic and urinary excretion of arsenic metabolites during boiler cleaning operations in a Slovak coal-fired power plant. AB - Little information is available on the relationship between occupational exposure to inorganic arsenic in coal fly ash and urinary excretion of arsenic metabolites. This study ws undertaken in a coal-fired power plant in Slovakia during a routine maintenance outage. Arsenic was measured in the breathing zone of workers during 5 consecutive workdays, and urine samples were obtained for analysis of arsenic metabolites--inorganic arsenic (Asi), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA)--prior to the start of each shift. Results from a small number of cascade impactor air samples indicated that approximately 90% of total particle mass and arsenic was present in particle size fractions >/= 3.5 micron. The 8-hr time-weighted average (TWA) mean arsenic air concentration was 48.3 microg/m3 (range 0.17-375.2) and the mean sum of urinary arsenic (SigmaAs) metabolites was 16.9 microg As/g creatinine (range 2.6-50.8). For an 8 hr TWA of 10 microg/m3 arsenic from coal fly ash, the predicted mean concentration of the SigmaAs urinary metabolites was 13.2 microg As/G creatinine [95% confidence interval (CI), 10.1-16.3). Comparisons with previously published studies of exposure to arsenic trioxide vapors and dusts in copper smelters suggest that bioavailability of arsenic from airborne coal fly ash (as indicated by urinary excretion) is about one-third that seen in smelters and similar settings. Arsenic compound characteristics, matrix composition, and particle size distribution probably play major roles in determining actual uptake of airborne arsenic. PMID- 9347900 TI - Exposure to regular gasoline and ethanol oxyfuel during refueling in Alaska. AB - Although most people are thought to receive their highest acute exposures to gasoline while refueling, relatively little is actually known about personal, nonoccupational exposures to gasoline during refueling activities. This study was designed to measure exposures associated with the use of an oxygenated fuel under cold conditions in Fairbanks, Alaska. We compared concentrations of gasoline components in the blood and in the personal breathing zone (PBZ) of people who pumped regular unleaded gasoline (referred to as regular gasoline) with concentrations in the blood of those who pumped an oxygenated fuel that was 10% ethanol (E-10). A subset of participants in a wintertime engine performance study provided blood samples before and after pumping gasoline (30 using regular gasoline and 30 using E-10). The biological and environmental samples were analyzed for selected aromatic volatile organic compounds (VOCs) found in gasoline (benzene, ethylbenzene, toluene, m-/p-xylene, and o-xylene); the biological samples were also analyzed for three chemicals not found in gasoline (1,4-dichlorobenzene, chloroform, and styrene). People in our study had significantly higher levels of gasoline components in their blood after pumping gasoline than they had before pumping gasoline. The changes in VOC levels in blood were similar whether the individuals pumped regular gasoline or the E-10 blend. The analysis of PBZ samples indicated that there were also measurable levels of gasoline components in the air during refueling. The VOC levels in PBZ air were similar for the two groups. In this study, we demonstrate that people are briefly exposed to low (ppm and sub-ppm) levels of known carcinogens and other potentially toxic compounds while pumping gasoline, regardless of the type of gasoline used. PMID- 9347901 TI - Birth weight reduction associated with residence near a hazardous waste landfill. AB - We examined the relationship between birth weight and mother's residence near a hazardous waste landfill. Twenty-five years of birth certificates (1961-1985) were collected for four towns. Births were grouped into five 5-year periods corresponding to hypothesized exposure periods (1971-1975 having the greatest potential for exposure). From 1971 to 1975, term births (37-44 weeks gestation) to parents living closest to the landfill (Area 1A) had a statistically significant lower average birth weight (192 g) and a statistically significant higher proportion of low birth weight [odds ratio (OR) = 5.1; 95% confidence interval (CI), 2.1-12.3] than the control population. Average term birth weights in Area 1A rebounded by about 332 g after 1975. Parallel results were found for all births (gestational age > 27 weeks) in Area 1A during 1971-1975. Area 1A infants had twice the risk of prematurity (OR = 2.1; 95 CI, 1.0-4.4) during 1971 1975 compared to the control group. The results indicate a significant impact to infants born to residents living near the landfill during the period postulated as having the greatest potential for exposure. The magnitude of the effect is in the range of birth weight reduction due to cigarette smoking during pregnancy. PMID- 9347902 TI - Total urinary follicle stimulating hormone as a biomarker for detection of early pregnancy and periimplantation spontaneous abortion. AB - Total concentrations of follicle stimulating hormone (FSH) were evaluated in daily urine samples from conceptive and nonconceptive menstrual cycles by measurement of the FSH beta subunit following treatment of the samples to dissociate the FSH heterodimer. Samples were self-collected by normal subjects during cycles in which daily blood samples also were obtained. Daily blood and urine specimens were collected prospectively from 10 subject in conceptive cycles, which led to normal pregnancies, and from 10 subjects with bilateral tubal ligations to provide control samples form nonconceptive cycles. Mean serum and urinary FSH concentration profiles wer parallel in both groups following ovulation and during he first 9 days of the luteal phase. Mean values for both serum and urinary FSH rose significantly above the postovulatory baseline by 10 12 days following the midcycle luteinizing hormone (LH) peak in nonconceptive cycles, but did not rise at any time following ovulation during conceptive cycles. Following regression analysis of the changing FSH concentration between days 9-14 post-LH surge in conceptive cycles, a slope of or = 65 years- United States, 1995. AB - In 1995, pneumonia and influenza together ranked sixth among the 10 leading causes of death in the United States. An estimated 90% of deaths caused by these illnesses occur among adults aged > or = 65 years. In addition, pneumococcal infections are the most common cause of bacterial pneumonia requiring hospitalization and account for an estimated 40,000 deaths annually in the United States. A national health objective for 2000 is to increase pneumococcal and influenza vaccination levels to > or = 60% for persons at high risk for complications from pneumococcal disease and influenza, including those aged > or = 65 years (objective 20.11). To estimate state-specific pneumococcal and influenza vaccination levels for persons aged > or = 65 years, CDC analyzed data from the 1995 Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the BRFSS findings, which indicate sustained increases in self reported coverage levels for pneumococcal and influenza vaccination among persons aged > or = 65 years; compares these findings with data from the 1993 BRFSS; and assesses progress toward the 2000 objective. PMID- 9347905 TI - Missed opportunities for pneumococcal and influenza vaccination of Medicare pneumonia inpatients--12 western states, 1995. AB - Invasive pneumococcal infection (i.e., bacteremia and meningitis) and influenza are important causes of morbidity and mortality among Medicare beneficiaries aged > or = 65 years. In the United States, the estimated annual incidence of pneumococcal bacteremia among persons aged > or = 65 years is 50-83 cases per 100,000 persons, and such infections are associated with a high case-fatality rate. Older persons account for >90% of influenza-related deaths, and Medicare costs for influenza-related hospitalizations can reach $1 billion each year. The Advisory Committee on Immunization Practices (ACIP) recommends that persons aged > or = 65 years receive at least one lifetime dose of pneumococcal vaccine and annual influenza vaccination and that hospitalization should be used as an opportunity to vaccinate. This report describes an assessment of the vaccination coverage of Medicare pneumonia patients who were admitted to hospitals in 12 western states from October 1994 through September 1995 (fiscal year 1995); the findings of this assessment indicate that the opportunity to provide pneumococcal vaccine was missed for up to 80% of those hospitalized at any time during the year, and the opportunity to provide influenza vaccine was missed for 65% of those who were admitted during October-December 1994. PMID- 9347906 TI - Health risk factor surveys of commercial plan- and Medicaid-enrolled members of health-maintenance organizations--Michigan, 1995. AB - Behavioral risk factors involving tobacco use, improper diet, physical inactivity, alcohol abuse, and motor-vehicle-related injury contributed to approximately 850,000 deaths in the United States in 1990. CDC's Behavioral Risk Factor Surveillance System (BRFSS) has been the primary means for tracking progress toward reduction of these and other health risks at the state level. This report summarizes the use of telephone surveys conducted by the Michigan Consortium for Quality Improvement in Health Care during April-August 1995 to evaluate the health-risk profiles of commercial plan- and Medicaid-enrolled health-maintenance organization (HMO) members in Michigan. The findings indicate that HMOs can use the BRFSS model to monitor health risks, health status, and use of preventive services among member groups. PMID- 9347907 TI - Poisonings associated with illegal use of aldicarb as a rodenticide -- New York City, 1994-1997. AB - Although rodenticides historically have been among the most toxic substances available to the public and have been implicated as agents in both unintentional and suicidal exposures, the anticoagulant agents currently in use, such as coumadin and their long-acting derivatives (e.g., brodifacoum), are relatively safe. In 1995, most persons who reported exposure to anticoagulant rodenticides did not develop symptoms or require specific therapy. However, during 1994-1997, the New York City Poison Control Center (NYCPCC) was consulted about 25 patients, primarily persons who had emigrated from the Dominican Republic, who had manifestations consistent with the cholinergic toxidrome, which is not characteristic of poisoning by the anticoagulant rodenticides, after ingesting a rodenticide known as Tres Pasitos ("Three Little Steps"). In each case, the product had been purchased at a neighborhood store for use as a household rodenticide. The Environmental Investigation Unit of the New York State Department of Environmental Conservation (NYDEC) investigated the poisoning incidents. Laboratory analysis indicated that the product contained the carbamate pesticide aldicarb (2-methyl-2-(methylthio)-propionaldehyde O-(methylcarbamoyl) oxime), which is not registered for use as a rodenticide in the United States. This report presents a detailed description of two of these cases and a summary of the remaining cases. PMID- 9347908 TI - Vaccination coverage by race/ethnicity and poverty level among children aged 19 35 months -- United States, 1996. AB - The Childhood Immunization Initiative (CII), implemented in 1993, is an intensive program to increase vaccination coverage among preschool-aged children and to reduce or eliminate vaccine-preventable diseases. In 1996, national coverage goals were achieved for 2-year-old children for the most critical doses of each routinely recommended vaccine. Disparities in vaccination coverage have been documented previously among different racial/ethnic groups. This report presents findings from CDC's National Immunization Survey (NIS), which document progress toward achieving the 1996 CII vaccination coverage goals by racial/ethnic group and by level of poverty. The findings indicate that, for each of five racial/ethnic groups, most of the national CII vaccination coverage goals were met and that, based on poverty level, all the goals were met for children living at or above the poverty level, and two of the five goals were met for children living below the poverty level. PMID- 9347909 TI - Current trends, IUD safety: report of a nationwide physician survey. 1974. PMID- 9347910 TI - Malaria surveillance -- United States, 1994. AB - PROBLEM/CONDITION: Malaria is caused by infection with one of four species of Plasmodium (i.e., P. falciparum, P. vivax, P. ovale, and P. malariae ), which are transmitted by the bite of an infective female Anopheles sp. mosquito. Most malarial infections in the United States occur in persons who have traveled to areas (i.e., other countries) in which disease transmission is ongoing. However, cases are transmitted occasionally through exposure to infected blood products, by congenital transmission, or by local mosquitoborne transmission. Malaria surveillance is conducted to identify episodes of local transmission and to adapt prevention recommendations. REPORTING PERIOD COVERED: Cases with onset of symptoms during 1994. DESCRIPTION OF SYSTEM: Malaria cases confirmed by blood smear are reported to local and/or state health departments by health-care providers and/or laboratories. Case investigations are conducted by local and/or state health departments, and the reports are transmitted to CDC through the National Malaria Surveillance System (NMSS), which was the source of data for this report. Numbers of cases reported through NMSS may differ from those reported through other passive surveillance systems because of differences in the collection and transmission of data. RESULTS: CDC received reports of 1,014 cases of malaria with onset of symptoms during 1994 among persons in the United States or one of its territories. This number represented a 20% decrease from the 1,275 cases reported for 1993. P. vivax, P. falciparum, P. malariae, and P. ovale accounted for 44%, 44%, 4%, and 3% of cases, respectively. More than one species was present in five persons (<1% of the total number of patients). The infecting species was not determined in 50 (5%) cases. The number of reported malaria cases in U.S. military personnel decreased by 86% (i.e., from 278 cases in 1993 to 38 cases in 1994). Of the U.S. civilians who acquired malaria during travel to foreign countries, 18% had followed a chemoprophylactic drug regimen recommended by CDC for the area to which they had traveled. Five persons became infected while in the United States; the infection was transmitted to two of these persons through transfusion of infected blood products. The remaining three cases, which occurred in Houston, Texas, were probably locally acquired mosquitoborne infections. Four deaths were attributed to malaria. INTERPRETATION: The 20% decrease in the number of malaria cases from 1993 to 1994 resulted primarily from an 86% decrease in cases among U.S. military personnel after withdrawal from Somalia. Because most malaria cases acquired in Somalia during 1993 resulted from infection with P. vivax, there was a proportionately greater decrease during 1994 in the number of cases caused by P. vivax relative to those caused by P. falciparum. ACTIONS TAKEN: Additional information was obtained concerning the four fatal cases and the five cases acquired in the United States. Malaria prevention guidelines were updated and distributed to health-care providers. Persons traveling to a geographic area in which malaria is endemic should take the recommended chemoprophylactic regimen and should use protective measures to prevent mosquito bites. Persons who have a fever or influenza-like illness after returning from a malarious area should seek medical care; medical evaluation should include a blood smear examination for malaria. Malarial infections can be fatal if not promptly diagnosed and treated. Recommendations concerning prevention and treatment of malaria can be obtained from CDC. PMID- 9347911 TI - Gastrulation and homeobox genes in chick embryos. AB - We review the early stages of chick embryogenesis, in particular the formation of the hypoblast, and the ingression of endoderm and mesoderm through the primitive streak. The formation of a trilaminar embryo during gastrulation is accompanied by the specification of body axes. The first axis is already present in the unfertilized egg and runs from the cytoplasmatic animal to the yolk rich vegetal pole. Already within the uterus a second axis conveys bilateral symmetry to the embryo. It extends from a dorsal/anterior to a ventral/posterior position. These axial poles segregate during gastrulation to form the classical coordinates, a dorsal-ventral and an anterior-posterior axis. The establishment of axes is accompanied by the expression of specific combinations of homeobox genes during gastrulation in the chick, as in other metazoa. We review the avian specific information and compare it with findings in other species. A combinatorial homeobox code for the specification of identities during development is discussed. PMID- 9347912 TI - The effect of dominant vestigial alleles upon vestigial-mediated wing patterning during development of Drosophila melanogaster. AB - The vestigial gene product is required for the completion of wing development in Drosophila melanogaster. In the absence of vestigial gene expression, cells within the larval wing and haltere imaginal discs fail to proliferate normally thus producing adults with severely reduced wings. Of a large number of vestigial mutations that have been characterized, only two are currently known to exist, vestigial(U) and vestigial(W), which manifest a significant dominant phenotype. Both are associated with chromosomal inversions that fuse the majority of the vestigial coding regions to other genes; mastermind in vestigial(U) and invected in vestigial(W) Examination of vestigial expression in the presence of these dominant alleles shows alterations in the disc-specific expression of vestigial during later stages of larval development. These patterning disruptions are specific to cells of the wing imaginal disc, as significant suppression of total levels of vestigial expression within entire larvae could not be detected. This dominant interference of vestigial patterning appears to be mediated in part by the vestigial coding sequences that are within the gene fusions. Further evidence that the dominant phenotype is the result of disrupted vestigial patterning comes from observations that the dominant alleles can be partially suppressed by mutations within the Drosophila-epidermal growth factor receptor gene. Mutagenesis of vestigial(U) and vestigial(W) produced a series of alleles with partially dominant phenotypes that restored various amounts of the adult wing. These phenotypes can be correlated with alterations in specific portions of the vestigial sequences associated with the dominant alleles. In the presence of these partially dominant alleles, wing imaginal discs have significantly more cells which express vestigial compared with the number associated with the original dominant phenotype. Additionally, eliminating some of the dominant effect causes alterations in the patterns of early stage apoptotic cell death associated with dominant vestigial alleles. Utilizing these new vestigial alleles, it is possible to correlate the consequence of altered vestigial expression to subsequent changes in patterning of the wing disc. PMID- 9347913 TI - Specification of endoderm in the sea urchin embryo. AB - Expression of the endoderm specific gene Endo16, was used to monitor endoderm specification in developmentally arrested and in dissociated embryos. beta-APN treatment halts gastrulation, however, in two species of sea urchins Endo16 mRNA is still expressed, suggesting that specification of the endodermal lineage has taken place in these developmentally arrested embryos. Endo16 mRNA is not expressed in embryos dissociated at the 4-8-cell stage unless they are reassociated shortly after the 16-cell stage. Interestingly, dissociation after the 16-cell stage also results in a lack of Endo16 expression. Lithium is unable to rescue Endo16 expression in these dissociated embryos. These results indicate that early signaling events mediated by cell-cell contact are required for the initial specification and maintenance of the endoderm in the developing embryo. PMID- 9347914 TI - Elements both 5' and 3' to the murine Hoxd4 gene establish anterior borders of expression in mesoderm and neurectoderm. AB - In this report, we show that a lacZ reporter spanning 12.5 kb of murine Hoxd4 genomic DNA contains the major regulatory elements controlling Hoxd4 expression in the mouse embryo. Mutational analysis revealed multiple regulatory regions both 5' and 3' to the coding region. These include a 3' enhancer region required for expression in the central nervous system (CNS) and setting the anterior border in the paraxial mesoderm, and a 5' mesodermal enhancer that directs expression in paraxial and lateral plate mesoderm. A previously defined retinoic acid response element (RARE) is a component of the 5' mesodermal enhancer. Our results support a model in which retinoic acid receptors (RARs) and HOX proteins mediate the initiation and maintenance of Hoxd4 expression. PMID- 9347915 TI - CNS midline to mesoderm signaling in Drosophila. AB - The dorsal median cells are unique mesodermal cells that reside on the surface of the ventral nerve cord in the Drosophila embryo. The Buttonless homeodomain protein is specifically expressed in these cells and is required for their differentiation. We have determined that proper buttonless gene expression and dorsal median cell differentiation requires signals from underlying CNS midline cells. Thus, dorsal median cells fail to form in single-minded mutants and do not persist in slit mutants. Through analysis of rhomboid mutants and targeted rhomboid expression, we also show that the EGF signaling pathway regulates the number of both the dorsal median cells, as well as a set of mesodermal cells that arise next to the midline and express the single-minded gene. Finally, wingless patched double mutants exhibit defects in the restriction of dorsal median cells to segment boundaries and alterations in CNS midline cell fates. Taken together, these data define a novel neuroectoderm to mesoderm signaling pathway and suggest that unique mesodermal cell types are specified by a combination of midline and segmental cues. PMID- 9347917 TI - Expression patterns of Brx1 (Rieg gene), Sonic hedgehog, Nkx2.2, Dlx1 and Arx during zona limitans intrathalamica and embryonic ventral lateral geniculate nuclear formation. AB - The Brx1 homeobox gene has been isolated and shown to be expressed in the zona limitans intrathalamica (ZLI) of the mouse embryo. Brx1 is a member of the Brx gene family and comprises the genes for Brx1a and Brx1b, which differ in the sequence in the region located on the 5'-terminal side of the homeobox. The complete amino acid sequences of the open reading frame of Brx1a and Brx1b were determined and each was found to be similar to that of Rgs, the mouse homologue of the Rieger syndrome associated human RIEG gene (RGS), to the extent that the sequence of Rgs has been clarified. Brx1 was strongly expressed in the mammillary area as well as in the ZLI of the mouse embryonic brain. Homologues of Brx1a and Brx1b were isolated in chick in which the expression of Brx1 in the ventral diencephalon was well conserved. The expression of Brx1 along with that of Sonic hedgehog (Shh), Nkx2.2, Dlx1 and Arx was examined at the time of the formation of ZLI in mouse embryos. The expression of Shh was initially noted in the ventricular zone of the presumptive ZLI and was then replaced by that of Brx1 at the time of radial migration of the neuroepithelial cells. Nkx2.2 was widely expressed in the ventricular zone of presumptive ZLI and also as a narrow band in the mantle zone. The expression of Dlx1 and Arx in the presumptive ventral thalamus extended as far as ZLI and overlapped with that of Brx1. The Dlx1- and Arx-expressing cells in ZLI, which extended towards the lateral (pial) surface of the diencephalic wall, differed from those expressing Nkx2.2 and Brx1. The embryonic ventral lateral geniculate nucleus present in the visual pathway was eventually formed from these cells. Each homeobox gene was also expressed regionally in the nucleus, suggesting that the nucleus is comprised of subdivisions. PMID- 9347916 TI - Somatic mesoderm differentiation and the development of a subset of pericardial cells depend on the not enough muscles (nem) locus, which contains the inscuteable gene and the intron located gene, skittles. AB - Not enough muscles (nem) mutants of Drosophila reveal defects in the development of embryonic muscles, a subset of pericardial cells, the CNS and derivatives of the PNS (Burchard, S., Paululat, A., Hinz, U. and Renkawitz-Pohl, R. (1995) The mutant not enough muscles (nem) reveals reduction of the Drosophila embryonic muscle pattern. J. Cell. Sci. 108, 1443-1454). The molecular analysis of the nem locus shows a complex genomic structure. One transcription unit was identified as inscuteable (insc). Within the first intron of insc we find another independent gene, skittles (sktl), which is not affected in nem mutants. insc transcripts are localised apically in neuroblasts and may prefigure the localisation of the protein. The skittles mRNA is ubiquitously distributed during early embryogenesis due to maternal contribution. Later, some enrichment of sktl is observed in the nervous system and the mesoderm. The muscle phenotype shows deletions as well as duplication of specific muscles which is reflected in a change of even-skipped (eve) and Kruppel (Kr) expressing cells. Our data suggest a role for insc in the specification process of a subset of muscle progenitors/founders. Furthermore, in insc mutants the eve expressing pericardial cells of the developing heart are significantly reduced in numbers. PMID- 9347918 TI - In vivo structure-function analysis of Drosophila Hairless. AB - Hairless plays an important role as the major antagonist in the Notch signalling pathway in Drosophila. It appears to be a direct inhibitor of the signal transducer Su(H). Hairless encodes a pioneer protein which was dissected in a structure-function analysis; a series of deletion constructs was tested for wild type and gain of function activity in the fly as well as for Su(H) binding. Thereby, the Hairless protein was subdivided into the absolutely essential Su(H) binding domain, similarly important N- and C-terminal domains and a central antimorphic domain. Therefore, Hairless protein might have additional functions apart from Su(H) binding and may antagonize Notch mediated cell-cell communication in a more complex way than currently anticipated. PMID- 9347919 TI - Effects of sodium sulfate on acute N-(3,5-dichlorophenyl)succinimide (NDPS) nephrotoxicity in the Fischer 344 rat. AB - The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) induces acute polyuric renal failure in rats. Results of previous studies have suggested that NDPS may induce nephrotoxicity via conjugates of NDPS metabolites. Thus, the purpose of this study was to examine if administered sodium sulfate could alter NDPS nephrotoxicity. Male Fischer 344 rats (four rats per group) were administered a single intraperitoneal (i.p.) injection of sodium sulfate (0.035, 0.07, 0.35 or 3.5 mmol/kg) or sodium chloride (7.0 mmol/kg) 20 min before NDPS (0.2, 0.4 or 0.8 mmol/kg) or NDPS vehicle (sesame oil, 2.5 ml/kg) and renal function monitored at 24 and 48 h. High dose sodium sulfate (3.5 mmol/kg) markedly attenuated NDPS nephrotoxicity, while sodium chloride had no effect on NDPS-induced renal effects. NDPS nephrotoxicity was also attenuated by a pretreatment dose of 0.35 mmol/kg sodium sulfate, while 0.07 mmol/kg sodium sulfate pretreatment potentiated NDPS 0.2 mmol/kg to produce nephrotoxicity without markedly attenuating NDPS 0.4 mmol/kg to induce renal effects. A dose of 0.035 mmol/kg sodium sulfate did not potentiate NDPS 0.2 mmol/kg to induce nephrotoxicity. These results suggest that sulfate conjugates of NDPS metabolites might contribute to NDPS nephrotoxicity. PMID- 9347922 TI - The cell-damaging effects of low amounts of homocysteine and copper ions in human cell line cultures are caused by oxidative stress. AB - In the present study, we have investigated the increase of cell protein and the concentration of glutathione, cysteine and homocysteine in cell culture systems (HeLa cell line) after addition of low amounts (100-500 micromol/l) of homocysteine and/or copper. The thiols and cell protein were determined in cell cultures with daily additions of new medium with and without homocysteine and/or copper ions for 3 days. The present study shows that extracellularly added homocysteine (500 and 2000 micromol/l) resulted in signs of cell toxicity (decreased intracellular glutathione level and/or retarded cell growth). After the addition of copper ions (10, 50 or 100 micromol/l), complex changes in the concentrations of thiols in cell cultures occurred but cell growth was normal. After the addition of both homocysteine and copper ions, changes similar to those seen with the addition of copper ions and homocysteine alone were noted. However, synergistic features after addition of 500 micromol/l homocysteine and 10 or 50 micromol/l of copper ions were a significantly retarded cell growth and decreased concentration of cellular glutathione. In HeLa cell lines with initial low cell density and in an endothelial cell line (ECV 304), even the presence of 100 micromol/l of homocysteine and 10 micromol/l of copper ions inhibited cell growth and decreased the cellular level of glutathione. Whilst the level of homocysteine in our 3-day cell-culture experiments is higher than the mild hyperhomocysteinemia thought to be atherogenic in humans (20-30 micromol/l), it is conceivable that over a longer time course (several decades), this mild hyperhomocysteinemia could be sufficient to induce cellular effects similar to those found in the present study, eventually leading to atherosclerosis. PMID- 9347920 TI - 1,3-Dinitrobenzene metabolism and toxicity in seminiferous tubules isolated from rats of different ages. AB - Previous in vivo studies in rats have shown that susceptibility to 1,3 dinitrobenzene (DNB)-induced testicular damage increases with age. The present study has used an in vitro approach to investigate the possibility that differences in testicular metabolism contribute to the age-related differences in toxicity. Seminiferous tubules were isolated from Sprague-Dawley rats (30, 75 and 120 days old) and incubated with 100 microM DNB for 22 h. Formation of metabolites and tubular levels of ATP and glutathione (GSH) were monitored over time. There was no difference in seminiferous tubule metabolic capacity among the three ages of rats examined. After 22 h of incubation with DNB, ATP levels were 20-30% of control and GSH levels were 70-90% of control, but neither parameter showed an age-related difference in decline. Based on these biochemical indicators of cell health, this study would suggest that the lack of testicular toxicity in young animals in vivo may be due to the previously described shorter half-life with consequent reduced exposure of the testis to DNB and that the age related increase in severity of lesion between 75 and 120 days of age cannot be explained by differences in tubular metabolism of DNB or whole-animal toxicokinetics. PMID- 9347921 TI - Change of cardiac beta-adrenoceptors in lead-exposed rats. AB - The effect of lead on cardiac beta-adrenoceptors was studied. Wistar rats used in these trials were divided into seven groups of ten animals each (A-G). Of these, groups B was given drinking water containing 0.01% lead acetate, group C 0.05%, group D 0.1%, group E 0.5%, group F 1% and group G 2% for a period of 60 days. Group A was given pure water. A radioligand-binding assay fulfilling strict criteria of receptor affinity and density was used to quantify cardiac beta adrenoceptors. Application of a trend test indicated that both blood and heart lead levels increased significantly from group A to group G (A < G), but that beta-adrenoceptor density decreased (G < A), whereas Kd did not vary among the seven groups. Linear regression analysis showed that decrease of cardiac beta adrenergic receptor density was closely related to elevation of blood and heart lead levels. The results show that lead exposure results in a reduction of cardiac beta-adrenoceptor density. PMID- 9347923 TI - Neuroprotection afforded by MK-801 against L-2-chloropropionic acid-induced cerebellar granule cell necrosis in the rat. AB - Administration of a single oral dose of 750 mg/kg L-2-chloropropionic acid (L CPA) to rats produces marked necrosis to the granule cell layer of the cerebellum by 48 h after dosing. Associated with the neuropathology the rats show locomotor impairment and a loss of body weight and a significant increase in cerebellar water and sodium content, indicating an oedematous reaction. Cerebellar aspartate and glutamate concentrations were reduced, while glycine and glutamine concentrations were increased after this treatment. Administration of the N methyl-D-aspartate (NMDA) receptor channel antagonist (5R,10S)-(+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d]cyclohepten-5,1 0-imine (MK-801), 30 min prior to L-CPA at a dose of 0.5, 1 or 5 mg/kg i.p. prevented the necrosis to the granule cell layer of the cerebellum and the signs of motor incoordination. Similarly there was no loss in cerebellar aspartate or glutamate concentration or increase in water or sodium content. Prior treatment with MK-801 at 0.1 mg/kg did not afford protection against the neurotoxicity. Post-treatment with 1 mg/kg MK-801 up to 1 h after administering L-CPA afforded complete neuroprotection, however if delayed until 2 or 6 h it gave only partial protection, and after 12 h it gave no protection. Administration of MK-801 alone at 5 mg/kg i.p., did not alter water content, sodium concentration, aspartate or glutamate concentrations in the cerebellum. In conclusion, we have shown that MK-801 given prior to and 1 h after L-CPA can afford complete neuroprotection, suggesting that a sub-population of NMDA receptors located on granule cells in cerebellum play a key role in mediating the selective toxicity of this chemical to the rat cerebellum. PMID- 9347924 TI - Mouse lung epithelial cell lines--tools for the study of differentiation and the neoplastic phenotype. AB - Several dozen lung epithelial cell lines have been established in culture over the past 20 years from normal lung explants and their spontaneous transformants, and from lung tumors that arose spontaneously or were induced with chemicals, viruses, or oncogenic transgenes. To provide information from which to choose appropriate lines for investigating problems in lung cell biology and pulmonary neoplasia, this review describes the origins of these lines and some of their characteristics. These include growth, morphology, tumorigenicity, ability to metastasize, xenobiotic metabolism, mutational status, signal transducing activities, cytogenetics, ability to form domes, and electric conductance. In addition to collecting this information in a single place for the first time, we describe previously unpublished apoptosis features of some of these lines. An increasing number of investigations are beginning to use these lines and this review contains references into 1997. PMID- 9347925 TI - Lack of correlation between hepatic prostaglandin concentrations and DNA synthesis after the administration of phenobarbital and the peroxisome proliferator ciprofibrate in rats. AB - Peroxisome proliferators are a class of chemicals that induce and promote hepatic tumors in rodents. These compounds are not genotoxic, and the mechanism by which they induce and promote tumors is poorly understood. Phenobarbital (PB) also is a hepatic tumor promoter that produces a different natural history than peroxisome proliferators during the promotion of hepatocarcinogenesis. In addition, opposite effects on hepatic eicosanoid concentrations have been demonstrated previously. In this experiment, we examined whether higher hepatic eicosanoid concentrations correlated with the induction of DNA synthesis after the administration of PB or the peroxisome proliferator ciprofibrate (CIP). PB (0.05% in diet) or CIP (0.01% in diet) was fed to rats from 1-10 days. For the rats treated with CIP, the peroxisomal enzyme fatty acyl-CoA oxidase increased gradually from day 1 to day 10. PB treated rats had a higher cytochrome P450 2B1/2 activity over the entire course of feeding. Hepatic prostaglandins E2 and F2alpha concentrations were significantly reduced in the rats treated with CIP, while no significant differences were seen between the control and PB-treated rats. DNA synthesis was increased in both PB-treated and CIP-treated rats. These results show that higher eicosanoid concentrations do not correlate with the induction of hepatic DNA synthesis by CIP or PB. PMID- 9347926 TI - Hemotoxicity of chlorpropham (CIPC) in F344 rats. AB - Chlorpropham[Isopropyl-N-(3-chlorophenyl)carbamate; CIPC] is a widely used sprout suppressant. Groups of ten male and ten female F344 rats were given 0, 7500, 15,000 or 30,000 ppm of CIPC in the diet for 13 weeks. Body weight gain of male and female rats in the 30,000 ppm-group was depressed. Spleen and liver weights of male and female rats in the treated groups were dose-dependently increased. Red blood cell count, hemoglobin concentration, hematocrit, mean corpuscular hemoglobin concentration and platelet count were decreased in male and female rats of the treated groups. Methemoglobin level, mean corpuscular volume and mean corpuscular hemoglobin were increased in male and female rats of the treated groups. Those hematological changes were dose-dependent and were marked in the 15,000 and 30,000 ppm-groups. White blood cell count of male and female rats in the 30,000 ppm-group were significantly higher than those of the control group. Congestion, increased hemosiderin deposition, increased extramedullary hemopoiesis, lymphoid atrophy and fibrosis were seen in spleen of male and female rats of all treated groups in a dose-dependent manner. Hemopoietic cell hyperplasia was marked in bone marrow of male and female rats in all treated groups. The results suggested that the erythrocyte is one of the primary targets of CIPC toxicity in rats. PMID- 9347927 TI - Comparison of the disposition of butadiene epoxides in Sprague-Dawley rats and B6C3F1 mice following a single and repeated exposures to 1,3-butadiene via inhalation. AB - 1,3-Butadiene (BD), a compound used extensively in the rubber industry, is a potent carcinogen in mice and a weak carcinogen in rats in chronic carcinogenicity bioassays. While many chemicals are known to alter their own metabolism after repeated exposures, the effect of exposure prior to BD on its in vivo metabolism has not been reported. The purpose of the present research was to examine the effect of repeated exposure to BD on tissue concentrations of two mutagenic BD metabolites, butadiene monoepoxide (BDO) and butadiene diepoxide (BDO2). Concentrations of BD epoxides were compared in several tissues of rats and mice following a single exposure or ten repeated exposures to a target concentration of 62.5 ppm BD. Female Sprague-Dawley rats and female B6C3F1 mice were exposed to BD for 6 h or 6 h x 10 days. BDO and BDO2 were quantified in blood and several other tissues following preparation by cryogenic vacuum distillation and analysis by multidimensional gas chromatography-mass spectrometry. Blood and lung BDO concentrations did not differ significantly (P < or = 0.05) between the two exposure regimens in either species. Following multiple exposures to BD, BDO levels were 5- and 1.6-fold higher (P < or = 0.05) in mammary tissue and 2- and 1.4-fold higher in fat tissue of rats and mice, respectively, as compared with single exposures. BDO2 levels also increased in rat fat tissue following multiple exposures to BD. However, in mice, levels of this metabolite decreased by 15% in fat, by 28% in mammary tissue and by 34% in lung tissue following repeated exposures to BD. The finding that the mutagenic epoxide BDO, which is the precursor to the highly mutagenic BDO2, accumulates in rodent fat may be important in assessing the potential risk to humans from inhalation of BD. PMID- 9347928 TI - Carbendazim and n-butylisocyanate: metabolites responsible for benomyl double action on cytochrome P450 in HepG2 cells. AB - Changes in the cytochrome P450 monooxygenase system were investigated in HepG2 cells treated for 24 h with 1.25, 2.5, 5, 10 and 20 microg/ml of carbendazim (MBC) and n-butylisocyanate (BIC), the principal benomyl metabolites. The results show that n-butylisocyanate leads to a decrease in both ethoxyresorufin deethylase (P4501A1) (EROD) and ethoxycoumarin deethylase (P4502B) (ECOD), whereas MBC has no effect on EROD and increases ECOD. The decrease in ECOD and EROD activities after BIC treatment can be attributed to the detrimental action of this substance. The MBC-induced increase in ethoxycoumarin can be considered an enzyme-specific inductive phenomenon. This hypothesis was confirmed by Western immunoblot analysis and treatment with actinomycin D 8 x 10(-4) microM: the first showed an increase in P4502B isoenzyme content and the second evidence of a partial block of the increase in ECOD activity induced by MBC. Given these results, MBC and BIC seem to be the metabolites responsible for the double opposite action of their parent compound benomyl. Data deriving from an equimolar mixture of the two metabolites suggest that benomyl activity on some cytochrome P450 isoenzymes is the result of a balance between the action of the single metabolites (Radice et al., 1996). PMID- 9347929 TI - Effects of occupational lead and cadmium exposure on some immunoregulatory cytokine levels in man. AB - The levels of serum interleukin-1beta (IL-1beta), interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-alpha) and gamma-interferon (gamma-IFN) were assessed in the workers who were occupationally exposed to lead and cadmium. The values were compared with the age-matched control group. Blood lead and cadmium levels were significantly raised. Our findings suggest that chronic lead and cadmium exposure in humans resulted in significant suppression of the serum IL-1beta level, but did not alter IL-2 and TNF-alpha levels. The gamma-IFN level was also reduced in lead workers. In contrast, a significant enhancement was observed in the cadmium-exposed group. We conclude from these results that lead and cadmium exposure at chronically high level may affect some cytokine levels in humans. PMID- 9347930 TI - An assessment of antigenic potential of beta-lactam antibiotics, low molecular weight drugs, using guinea pig models. AB - Allergic reactions are among the common adverse effects in humans. However, it is widely assumed that there are practically no reliable animal models for preclinical tests of low-molecular weight drugs that are available to predict such reactions. This study was designed to compare the detecting ability of test methods for antigenic potential of eight beta-lactam antibiotics with which allergic outcome has been reported in humans. The tests included active systemic anaphylaxis (ASA), delayed type skin reaction (DSR), maximization test (GPMT) in guinea pigs sensitized with antibiotics emulsified with Freund's complete adjuvant, passive cutaneous anaphylaxis (PCA) and enzyme-linked immunosorbent assay (ELISA) as serological tests. PCA and ELISA though using protein-conjugates as detecting antigens, especially ELISA, showed positive reactions with relatively high incidence. On the other hand, GPMT was the most sensitive method to detect antigenic potential of antibiotics despite the use of antibiotics alone for sensitizing and challenging phases. It is suggested that GPMT can be considered the most reliable method in preclinical testing. PMID- 9347931 TI - The results of sensitisation testing of five metals in a mouse model. PMID- 9347933 TI - The density of 5-hydoxytryptamine2A receptors in forebrain is increased at pro oestrus in intact female rats. AB - We have shown previously that in ovariectomised rats, oestradiol-17beta, in its positive-feedback mode for luteinizing hormone (LH) release, induces a significant increase in the density of 5-hydroxytryptamine2A (5-HT2A) receptors in the forebrain. Here we investigated whether there are any changes in 5-HT2A receptor density in relation to the spontaneous surge of oestradiol-17beta in female COB Wistar rats between dioestrus and pro-oestrus. Using [3H]RP62203 binding and autoradiography, we found that 5-HT2A binding sites were significantly increased at 1630-1800 h on pro-oestrus compared with 0900-1130 h on dioestrus in frontal and cingulate cortex, olfactory tubercle and nucleus accumbens. The densities of 5-HT2A binding sites in male rats were similar to dioestrous female values in cortex, and to pro-oestrous female values in nucleus accumbens. The changes in the density of 5-HT2A binding sites in the forebrain of female rats may be relevant to oestrogen effects on mood and mental state. PMID- 9347932 TI - A novel pathogenic mutation (Leu262Phe) found in the presenilin 1 gene in early onset Alzheimer's disease. AB - Several mutations causing early-onset familial Alzheimer's disease (AD) have been detected in the presenilin 1 (PS-1) gene. Pathogenic mutations have also been described in an homologous gene, presenilin 2 (PS-2). In order to screen for mutations in these genes, cDNA samples from early-onset AD cases were analysed, using single strand conformation polymorphism (SSCP) and direct cDNA sequencing. Two missense mutations in the PS-1 gene were detected, a previously unidentified amino acid substitution Leu262Phe and an earlier reported amino acid substitution Glu318Gly. No disease-related mutations were found in the PS-2 gene. PMID- 9347934 TI - Syrian hamster prion protein (PrP(C)) is expressed in photoreceptor cells of the adult retina. AB - PrP(C), the cellular isoform of the prion protein (PrP) serves as a precursor to abnormal PrP isoforms which accumulate in diseases such as scrapie in sheep, and Creutzfeldt-Jakob disease in humans. Since prions can replicate in photoreceptors we surmised that PrP(C) must be expressed in these cells. Accordingly, monoclonal antisera directed against two epitopes of hamster PrP(C) produced retinal immunostaining in hamsters, and in mice bearing a hamster PrP transgene. Immunostaining was most prominent in the inner and outer segments of rod photoreceptors, coinciding with the earliest site of pathologic changes in scrapie-infected hamsters. These data define PrP(C) expression in an experimentally-accessible population of neurons and suggest that the retina may comprise a useful system for studying the biology of wild-type and mutant prion proteins. PMID- 9347935 TI - Prion protein (PrP) is not involved in the pathogenesis of spongiform encephalopathy in transgenic mice expressing interleukin-6 in the brain. AB - Transgenic mice expressing interleukin-6 (IL6) in the brain exhibit gliosis, spongiosis and neuronal loss. Based on previous findings, we hypothesized that IL6 could upregulate the prion protein (PrP) gene in the central nervous system (CNS) of these mice. Western and Northern blot analysis showed that PrP protein and mRNA levels were comparable to control levels. Furthermore, ultrastructural characterization revealed that spongiosis was actually located in astrocytes. These results indicate that IL6 does not upregulate the cerebral PrP expression in this animal model and that profound astrocytic alterations precipitate the neuronal degeneration observed. PMID- 9347936 TI - No association between an intronic polymorphism in the presenilin-1 gene and Alzheimer's disease. AB - Mutations in the presenilin-1 (PS-1) gene are believed to be responsible for the majority of familial early-onset Alzheimer's disease (AD). The finding of an intronic polymorphism in the PS-1 gene prompted an investigation into its relevance in AD. An association between homozygosity for the most common allele (allele 1) in this intronic polymorphism and late-onset AD has been shown and has been confirmed by others though some studies do not support these findings. We genotyped a large series of sporadic AD cases (n = 120) and age-matched controls (n = 108) for this intronic polymorphism. We then compared both the frequency of allele 1 and allele 1 homozygosity between the AD group as a whole and in early onset (n = 26) and late-onset (n = 94) groups with age-matched control groups (n = 29 and n = 79, respectively). No increase in the frequency or homozygosity of allele 1 in either the AD group as a whole, or when divided into late- and early onset cases was found. Increases in the frequency of allele 1 homozygotes and in the number of non-apolipoprotein E epsilon4 carrying allele 1 homozygotes/heterozygotes was demonstrated in the early-onset AD cases although these values did not reach significance. We conclude that there is no relationship between this intronic polymorphism in the PS-1 gene and AD in the homogenous population genotyped in this study. PMID- 9347937 TI - PET study of the localization and laterality of lingual somatosensory processing in humans. AB - Regional cerebral blood flow (rCBF) was measured using positron emission tomography (PET) during four tasks in right-handed volunteers with eyes closed: resting, protruding the tongue, stroking the left side of the protruding tongue, and stroking the right side of the protruding tongue. The primary somatosensory tongue representation (S1) mapped to the contralateral central sulcus (Brodmann (BA) 3/4) at approximately 28 mm above the intercommissural plane. Of note, stimulation of the left side of the tongue produced also an ipsilateral S1 response. Analysis of variance (ANOVA) of rCBF at S1 across all four conditions yielded only a significant effect for tongue stimulation, with no effect of laterality; the usually large asymmetries (contralateral >> ipsilateral) in S1 did not surface. We hypothesize that this atypical activation pattern arises from the tongue's specialization for language. PMID- 9347938 TI - Opioid-related changes in nociceptive threshold and in tissue levels of enkephalins after target disruption of the gene for neutral endopeptidase (EC 3.4.24.11) in mice. AB - Neutral endopeptidase EC 3.4.24.11 (NEP) is localized in peptidergic neurons and various colocalized peptides or other humoral mediators may serve as substrates. Target disruption of the NEP gene was reported to enhance the lethal response to endotoxin shock in mice. We examined thermonociceptive thresholds and enkephalin (ENK) tissue levels in transgenic NEP (-/-) and control wild type NEP (+/+) mice. Hot plate (52 degrees C) latency was 13.1 +/- 1.4 s in NEP (+/+) mice (n = 16) while latency increased significantly (P = 0.031) to 17.7 +/- 1.6 s in NEP (-/-) mice. Naloxone (10 mg/kg) had no effect on hot plate latency in NEP (+/+) mice (12.5 s, n = 8), but significantly decreased the latency in NEP (-/-) mice compared to untreated NEP (-/-) deficient mice (10.5 s, n = 8). Morphine (3 or 10 mg/kg) analgesic response was similar in knockout mice and wild type mice. Methionine-ENK (MET-ENK) and leucine-ENK (LEU-ENK) levels were determined in extracts from cortex, brain stem, hypothalamus, striatum, spinal cord, trigeminal ganglion and heart in treated and untreated mice. ENK-levels varied in a regionally-dependent manner and were significantly decreased in hypothalamus and spinal cord. We conclude that deletion of the NEP gene results in an opioid related increase in thermonociceptive threshold. Regional differences in opioid metabolism indicate that NEP evokes tissue-specific patterns of ENK-regulation. NEP selectively controls opioid biosynthesis in hypothalamus and spinal cord presumably by feedback regulation. PMID- 9347939 TI - Metabotropic glutamate autoreceptors on nerve terminals of crayfish muscle depress or facilitate release. AB - In lobster and crayfish neuromuscular junctions superfusion of the excitatory transmitter L-glutamate (L-Glu) has been shown to depress release from motor axon terminals. In crayfish the effect depended on the level of depolarization of the terminal, Glu facilitating release when large depolarizing stimuli were applied. The presynaptic inhibition by Glu could be blocked only with a combination of blockers of Glu channels of the AMPA and N-methyl-D-aspartate (NMDA) types. We report that the effects of Glu can be mimicked by superfusion of (1S,3R)-1 aminocyclo-pentane-1.3-dicarboxylic acid (t-ACPD), an agonist of vertebrate metabotropic Glu-receptors. However, in the recent run of experiments the majority of terminals reacted with facilitation of release on superfusion by Glu or t-ACPD, even when the depolarizing stimuli to the terminal or the control rate of release were low. Average facilitation by 5 microM Glu was by a factor of 4.5. Facilitory responses were not blocked by the combination of APV and CNQX, and thus two types of metabotropic receptors seem to be present, their proportion varying for unknown reasons. PMID- 9347940 TI - Nociceptin and its receptor in guinea-pig sympathetic ganglia. AB - The occurrence and distribution of nociceptin, a 17-amino acid peptide with structural similarity to dynorphin A, and its receptor, opioid receptor-like-1 (ORL1) receptor, were investigated in the guinea-pig sympathetic nervous system by means of immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR), respectively. Immunofluorescence revealed varicose nociceptin immunoreactive axons and some paraganglionic cells in prevertebral (coeliaco superior mesenteric, inferior mesenteric), but not in paravertebral (superior cervical, stellate, lumbar chain) sympathetic ganglia. Messenger RNA for the ORL1 receptor, however, was detected by RT-PCR in both para- and prevertebral ganglia. The findings suggest participation of the nociceptin/ORL1 receptor signalling pathway in processing of information within prevertebral ganglia, and a general responsiveness of sympathetic neurons to nociceptin. PMID- 9347941 TI - Involvement of alpha2-adrenoceptors in mediating sympathetic excitation of injured dorsal root ganglion neurons in rats with spinal nerve ligation. AB - The present study examined the effects of sympathetic stimulation on the activity of primary afferent neurons that had peripheral axons being injured previously by a spinal nerve ligation. About 22% of afferents with injured fibers that showed spontaneous discharge were excited by sympathetic stimulation or systemic injection of adrenaline. Most sympathetically-excited afferent neurons had axons that conducted in the A-fiber range. This sympathetically-evoked afferent excitation was not affected by cutting the spinal nerve at a place close to the dorsal root ganglion (DRG). Yohimbine, alpha2-antagonist, suppressed sympathetically-evoked afferent excitation which was not affected by alpha1 antagonist prazosin. Clonidine, alpha2-agonist, exerted an excitatory effect, whereas alpha1-agonist phenylephrine had no effect on the activity of afferents with injured fibers. No afferent fibers in control preparations responded to sympathetic stimulation. The results suggest that after a spinal nerve ligation, injured DRG neurons with fast-conducting fibers become sensitive to sympathetic activity via activation of alpha2-adrenoceptors. PMID- 9347942 TI - Ciliary neurotrophic factor stimulates in vivo myotube formation in mice. AB - It is known that ciliary neurotrophic factor (CNTF) administration reduces the atrophy observed with denervation, suggesting its role as a trophic factor for muscle cells. At the present, we studied the effects of 'in vivo' CNTF administration on the regenerative capacity of skeletal muscle fibres. Adult mice had their extensor digitorium longus muscle subjected to a denervation devascularization lesion. CNTF (0.5 ng/microl) was administered using osmotic pumps implanted subcutaneously in unrestrained mice. CNTF was delivered into the muscle's region at a rate of 1 microl/h from 1 to 8 days after denervation. The results show that CNTF increased the number of regenerating myofibres by day 4. From day 7 on, the values seen on control and CNTF-treated groups were not significantly different. Our results show that 'in vivo' CNTF administration accelerates myotube differentiation. PMID- 9347943 TI - Kinetics of nigral degeneration in a chronic model of MPTP-treated mice. AB - The chronic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model we have developed in monkey reproduces all the cardinal features of Parkinson's disease, and, in particular the characteristic slow evolution of clinical signs. We still know little, however, of the kinetics of the nigral degeneration induced. This present study charts the progressive destruction of tyrosine hydroxylase immunoreactive neurones in mice treated daily with low doses of MPTP for 20 days. Our results show that the neuronal death rate is initially high, subsequently decreases, and stabilizes. This new protocol thus mirrors closely the pattern of evolution assumed to be that of Parkinson's disease and should prove useful for studies on neuroprotection and compensatory mechanisms. PMID- 9347944 TI - Paradoxical lateral suppression in the dolphin's auditory system: weak sounds suppress response to strong sounds. AB - A paradoxical phenomenon was found in the auditory system of dolphins: weak sounds suppressed the brain responses to much stronger sounds. This occurred when the brain evoked potentials to rhythmic sound amplitude modulations were recorded. The response was markedly suppressed by addition of another sound of higher frequency and down to 40 dB lower intensity than the amplitude-modulated signal. Only the sustained rhythmic response was suppressed while transient on response was not, thus indicating that the suppression influenced the ability of evoked potentials to follow rapid amplitude modulations. This prevents weak sounds from being masked by stronger ones. It may help a dolphin to perceive weaker echo-signals in the background of stronger emitted pulses. PMID- 9347945 TI - The effect of stereocilia bundle position on receptor potential characteristics of isolated outer hair cells in the guinea pig. AB - The receptor potential as a function of pre-stimulus position of the stereocilia was studied with the whole cell patch-clamp technique in isolated outer hair cells (OHC) during direct mechanical stimulation of the stereocilia (n = 6, cells ranging from 70-87 microm length). The amplitude and frequency of the stimulation was 1 microm and 100 Hz. Controlled pre-stimulus stereocilia displacements in the excitatory direction were followed by sustained membrane depolarization and a diminution of the DC component of the receptor potential. Spectral analysis has shown that recordings with most diminished DC components had a pronounced fundamental while the other harmonics were suppressed, indicating the linearization of the receptor potential. The observed non-lineararities depended on the resting position of the stereocilia and seemed to be determined by the operating range of the transduction process. PMID- 9347946 TI - The cloned rat hydrolytic enzyme responsible for the breakdown of anandamide also catalyzes its formation via the condensation of arachidonic acid and ethanolamine. AB - Anandamide amidase is the hydrolytic enzyme responsible for the breakdown of anandamide, an endogenous cannabimimetic, to arachidonate and ethanolamine. Another enzymatic activity called anandamide synthase catalyzes the reverse reaction, that is the condensation of arachidonate and ethanolamine. Using a recently cloned rat fatty acid amidohydrolase (FAAH), we tested the hypothesis that the synthase and the amidase activities are catalyzed by the same enzyme. Untransfected and vector transfected (pcDNA3) COS-7 cells did not express detectable levels of either the amidase or synthase. However, when COS-7 cells were transiently transfected with a rat FAAH pcDNA3 construct, both amidase and synthase were concomitantly expressed. These results indicate that the enzymatic formation of anandamide from arachidonic acid and ethanolamine can be mediated by anandamide amidase acting in the reverse direction. The FAAH transfected cells expressed higher levels of enzyme than either rat brain homogenates or neuroblastoma cells in culture. Furthermore, the reaction rate for the amidase in FAAH transfected COS-7 cells, neuroblastoma cells and brain homogenate was always greater than the synthase reaction. These studies raise the question if this synthase reaction serves any physiological role, especially in view of the evidence that anandamide can be formed by a different pathway. PMID- 9347947 TI - Tau immunoreactivity in glial cytoplasmic inclusions in multiple system atrophy. AB - In order to clarify the manner and significance of tau expression in glial cytoplasmic inclusions (GCIs), ubiquitinated oligodendroglial abnormal structures in multiple system atrophy (MSA), an immunohistochemical study was carried out in the lesions of the pontine nuclei of 10 cases of MSA using antibodies against various epitope locations of tau protein. As a result, tau-2 was constantly but weakly positive in ubiquitinated GCIs in each case (from 28.6 to 66.7%). However, tau-2-immunoreactivity in GCIs was not correlated to the density of ubiquitin positive GCIs or preserved pontine neurons. Antibodies against tau proteins of N terminal or C-terminal failed to label GCIs, although a few number of GCIs were occasionally positive for tau-1 after dephosphorylation. In comparison with the knowledge on tau-immunoreactivity of coiled bodies (CBs) in oligodendroglia in progressive supranuclear palsy (PSP) or corticobasal degeneration, GCIs are quite different from CBs which have a wide range of epitope location of tau proteins, including N-terminal and C-terminal. This study suggests that expression of tau proteins in GCIs is not related to the essential neurodegenerative process in MSA but induced by non-specific stress in oligodendroglia, unlike CB in various 'tau diseases' such as PSP. PMID- 9347948 TI - Cholecystokinin (CCK-8) modulates vesicular release of excitatory amino acids in rat hippocampal nerve endings. AB - The modulation of endogenous amino acid transmitter release by the sulphated octapeptide cholecystokinin (CCK-8S) was investigated in purified rat hippocampal synaptosomes. In the presence of extracellular Ca2+, CCK-8S increased the basal release of glutamate, but not of aspartate and GABA. In addition, CCK-8S dose dependently increased the KCl-evoked Ca2+-dependent release of both glutamate and aspartate to about 1.4-fold at concentrations > or = 0.5 microM. CCK-8S did not change the KCl-evoked Ca2+-dependent GABA release, not even in the presence of the GABA uptake carrier blocker N-(4,4-diphenyl-3-butenyl)-3-piperidine carboxylic acid 89976-A (SK&F89976-A; 10 microM). The CCKB receptor antagonist L365,260 (1 microM) blocked the CCK-8S-induced release of glutamate by 70%, and of aspartate by 100%. In conclusion, CCK stimulates exocytosis of excitatory amino acids in rat hippocampus by activating a low-affinity presynaptic CCK receptor, presumably of the B-subtype. However, CCK does not modulate the release of GABA, which has been reported to be colocalized with this peptide. PMID- 9347949 TI - Beta-amyloid induced increase in choline flux across PC12 cell membranes. AB - Beta-amyloid peptide is the main constituent of senile plaques and is implicated in the pathogenesis of Alzheimer's disease. It has been shown to be both neurotoxic and neurotrophic in vivo, and its effects have been suggested to be mediated in part by alterations in membrane transport. In the present study, we investigated the effect of beta-amyloid (1-40) on choline transport in cultured PC12 cells. We found that exposure to 46 or 92 microM beta-amyloid (1-40) increased [14C]choline flux in PC12 cells in a concentration-dependent manner, whereas exposure to reverse sequence beta-amyloid (40-1) had no effect. If there is a similar effect in vivo, the increased beta-amyloid dependent permeability to choline could lead to depletion of cellular choline stores and could contribute to the selective vulnerability of cholinergic neurons in Alzheimer's disease. PMID- 9347950 TI - Regulation of leukocyte adhesion molecules CD11b/CD18 and leukocyte adhesion molecule-1 on phagocytic cells activated by malaria pigment. AB - There is increasing evidence that inappropriate immune activation induced by parasite products occurs in malaria disease. To further elucidate the role of Plasmodium falciparum-derived products on host immune activation, we studied the expression of leukocyte adhesion molecules (CD11b/CD18 and LAM-1) on neutrophils and monocytes in response to malaria pigment using flow cytometry. Exposure of leukocytes to isolated malaria pigment derived from ruptured schizonts resulted in significant up-regulation of CD11b/CD18 expression and down-regulation of LAM 1 on both neutrophils and monocytes. In contrast, culture supernatants (pigment free) from ruptured schizonts did not alter the expression of CD11b/CD18 and LAM 1. The increase of CD11b/CD18 and the loss of LAM-1 expression occurred simultaneously with the earliest response detected at 10 min and a plateau reached by 60 min. The effect of malaria pigment on leukocyte adhesion molecules was inhibited by EDTA in a dose-dependent manner. Phagocytosis of malaria pigment was also suppressed by EDTA. This observation suggests that phagocytosis of malaria pigment may be a prerequisite for the effect of malaria pigment on the regulation of CD11b/CD18 and LAM-1 expression. Regulation of leukocyte adhesion molecules through up-regulation of CD11b/CD18 and down-regulation of LAM-1 by malaria pigment could promote leukocyte adherence to endothelium in vivo. This increased adherence of malaria pigment-activated leukocytes might induce cytokine (tumor necrosis factor alpha and interleukin-1beta)-mediated increases in capillary permeability resulting in local tissue edema, and a cytokine-mediated increase in adhesion molecule expression causing vascular clogging by adherent red blood cells, and in severe disease by adherent leukocytes. PMID- 9347951 TI - Receptor-specific adhesion and clinical disease in Plasmodium falciparum. AB - One important factor in the virulence of infections with Plasmodium falciparum is the adherence of infected erythrocytes to small vessel endothelium. In infections that lead to serious, life-threatening disease accumulation of large numbers of infected cells in particular organs is thought to lead to organ dysfunction or failure. This is of particular relevance when the affected organ is the brain, leading to the development of cerebral malaria. Many different endothelial receptors for infected red blood cells have been identified. Some receptors such as CD36 and thrombospondin are used by all parasite isolates, whereas others such as intercellular adhesion molecule-1 (ICAM-1) or vascular cell adhesion molecule (VCAM) are used by a subset of field and laboratory isolates. While it has been speculated that the ability to bind or affinity of binding to a particular endothelial receptor may be related to the pattern of disease, only studies with limited numbers of patients have been carried out to date and these have been in general inconclusive. Here we have taken parasite isolates from 150 patients with defined clinical syndromes as well as isolates from 50 healthy but parasitized community controls and quantitatively assessed their binding to purified endothelial receptors in vitro. Our results show that disregarding the level of adhesion, all parasites bind to CD36, most bind to ICAM-1, few bind to VCAM, and almost none bind to E-selectin. In assessing the degree of binding we show that 1) binding to all receptors was reduced in parasites taken from severely anemic patients; 2) binding to CD36 is identical in parasites from cerebral malaria patients and community controls but slightly elevated in parasites from nonsevere cases; and 3) binding to ICAM-1 is highest in cerebral malaria patients. Because rosette formation by uninfected cells has also been a phenotype associated with disease severity and one that may interfere in vitro with receptor binding, we also assessed rosette formation in all isolates. In this study the highest level of rosette-forming parasites was found in the anemic group and not the cerebral malaria group. Stratifying the data for the frequency of rosette formation showed that the above results were not significantly altered by this phenomenon. Our data are not consistent with a role for binding to CD36 in the development of severe disease but show an association between the degree of binding to ICAM-1 and clinical illness in nonanemic patients. PMID- 9347952 TI - Comparison of whole blood and serum levels of mefloquine and its carboxylic acid metabolite. AB - Because of the widespread presence of chloroquine-resistant Plasmodium falciparum malaria, mefloquine is now the recommended drug of choice for long-term malaria prophylaxis in these areas. Although several studies have compared plasma and whole blood concentrations of either mefloquine or its carboxylic acid metabolite, we report the first comparison of serum and whole blood levels in 86 Dutch marines taking 250 mg of mefloquine weekly for 18 weeks while deployed in western Cambodia. All samples were taken during steady-state and at 42-48 hr after the most recent dose. The concentration of mefloquine in serum (mean = 979 ng/ml) was significantly greater than in whole blood (mean = 788 ng/ml) (P < 0.00001, by paired t-test) with an overall mean ratio of 1.28. The concentration of the metabolite in serum (mean = 3,039 ng/ml) was also significantly greater than in whole blood (mean = 1,390 ng/ml) (P < 0.00001, by paired t-test) with an overall mean ratio of 2.25. These findings are similar to previous reports of plasma-to-whole blood levels. Furthermore, we report that the within-individual ratios of the metabolite concentration to the mefloquine concentration were also found to be significantly different in serum (3.79; P < 0.00001, by paired t test) and in whole blood (2.02; P < 0.00001, by paired t-test). Appropriate attention must be given to these differences when comparing serum and whole blood concentrations of either mefloquine or its metabolite to avoid misinterpretation of their respective levels. Also, the determination of the relative mefloquine ratios in various blood fluids, as well as the documentation of the metabolite levels and their ratios, is critical to the appropriate interpretation of both chemoprophylaxis and chemotherapy, especially in the presence of resistant strains. PMID- 9347953 TI - Pharmacokinetics of liposome-encapsulated meglumine antimonate after intramuscular and subcutaneous administration in dogs. AB - Controlling canine leishmaniasis may reduce the incidence of human leishmaniasis, which affect immunocompromised persons, especially those with human immunodeficiency virus infection. Thus, the pharmacokinetics of liposome encapsulated meglumine antimonate (LMA) in dogs was studied after intramuscular (I.M.) and subcutaneous (S.C.) administration. Serum concentration-time data for both forms of administration were best described by a triexponential open model. The absorption phase showed statistically significant differences between I.M. and S.C. administrations (K01(I.M.) = 0.046/min, K01(S.C.) = 0.025/min). The first phase of decrease of plasma concentrations showed a longer half-life for S.C. than for I.M. administration, with the delay being caused by the slow absorption process after S.C. injection. Mean terminal phase half-lives after administration of I.M. and S.C. were 904.1 min and 637.4 min, respectively. Peak plasma concentrations after administration of I.M. (Cmax = 43.8 microg/ml) and S.C. (Cmax = 24.9 microg/ml) were detected at 42.8 min and 79.8 min, respectively. Urinary excretion of antimony for both routes surpassed 80% during the first 6 hr, with the rest of the drug being excreted slowly over the following 18 hr. The results obtained with this formulation suggest that for treating canine leishmaniasis, it would be more advisable to inject LMA intramuscularly if we assume that the significantly higher Cmax observed after I.M. administration is more relevant to dog's clinical outcome than is maintenance of concentrations over longer periods. PMID- 9347954 TI - Specific inhibitory effect of 3-deazaneplanocin A against several Leishmania mexicana and L. braziliensis strains. AB - The growth inhibitory effect of 3-deazaneplanocin A (c3NpcA) was tested against some pathogenic members of the family of American Trypanosomatidae. Under our culture conditions, c3NpcA displayed a strongly and uniformly leishmanistatic effect on all 23 American Leishmania (L. mexicana and L. brasiliensis) strains in the study (mean dose producing 50% inhibition compared with control parasite growth [ID50] = 96 ng/ml, 0.32 microM), but showed no inhibition against the several T. cruzi and T. rangeli strains tested with concentrations up to 10,000 ng/ml. This compound also induced a substantial expansion of the intracellular pools of both S-adenosylhomocysteine (AdoHcy) and S-adenosylmethionine (AdoMet), as well as a significant diminution of the AdoMet:AdoHcy ratio. Strong AdoHcy hydrolase activity was detected in American Leishmania promastigotes. At at a dose of 200 ng/ml, c3NpcA inhibited S-adenosyl-L-3H-methylmethionine and 3 thymidine incorporation by promastigotes after four days incubation in the presence of the drug. At a dose of 100 ng/ml, c3NpcA eliminated approximately 56% of the L. mexicana and L. brasiliensis from infected human macrophages, compared with simultaneously cultivated controls. Two schedules of 10 consecutive intraperitoneal injections of c3NpcA, with doses ranging from 0.5 to 1.5 mg/kg/day, significantly reduced development of cutaneous leishmanial infection produced in inbred BALB/c mice by L. b. guyanensis inoculation, although a few parasites remained at the inoculation site. PMID- 9347955 TI - Strongyloides hyperinfection in a renal transplant recipient receiving cyclosporine: possible Strongyloides stercoralis transmission by kidney transplant. AB - Strongyloides hyperinfection and dissemination are recognized complications in kidney allograft recipients; however, the development of strongyloidiasis in renal transplant recipients receiving cyclosporine A (CyA) has not been described, nor has the development of strongyloidiasis in other organ transplant recipients. The former observation has been attributed to the antiparasitic activity of CyA seen in animal studies; the latter has no explanation yet. We report the first case of Strongyloides hyperinfection in a renal transplant patient occurring immediately after CyA was discontinued. From the unique characteristics of this case, it appears that the anti-Strongyloides activity of CyA in animals may also be found in humans. PMID- 9347956 TI - Rapid characterization of genetic diversity among twelve dengue-2 virus isolates by single-strand conformation polymorphism analysis. AB - Single-strand conformation polymorphism (SSCP) analysis was used to characterize genetic polymorphisms among 12 isolates of dengue-2 virus, which were previously genetically characterized by RNase T1 oligonucleotide mapping and by sequencing the viral envelope (E) gene. Specific cDNA fragments from the dengue-2 isolates were amplified by the reverse transcriptase-polymerase chain reaction. The viral E, premembrane (prM), and nonstructural 5 (NS5) gene cDNAs of 291 basepairs (bp), 291 bp, and 201 bp, respectively, were denatured, rapidly chilled to promote intrastrand reassociation, electrophoretically separated on nondenaturing polyacrylamide gels, and SSCP patterns were observed by silver staining. The SSCP analysis revealed polymorphisms among a number of dengue-2 isolates from the same topotype, and these were markedly different between isolates of different topotype (distinct genetic group). Comparison of nucleotide sequence and SSCP analyses of the 291-bp E cDNA demonstrated that virus isolates that produced identical SSCP patterns contained 0-7 nucleotide substitutions, whereas isolates that showed different SSCP patterns contained 4-25 nucleotide substitutions. Positive predictive value and negative predictive value as measures of certainty for predicting identical and different sequences were 26% and 100%, respectively. The SSCP patterns of the 12 dengue-2 isolates suggested greater genetic variation in the prM gene region than in either the E or NS5 gene regions. The SSCP analyses should allow easy, sensitive, and rapid screening of dengue viruses isolates and the assessment of variation at a number of sites in the virus genome. Additionally, SSCP screening of dengue-2 virus for genetic variability may reveal the introduction of new viral genotypes in a given geographic area. These genetic variants of the virus could serve as markers of the epidemic potential of the virus strain. PMID- 9347957 TI - Short report: Leishmania DNA in Phlebotomus and Sergentomyia species during a kala-azar epidemic. AB - The presence of Leishmania donovani DNA in sand flies caught in Indian kala-azar patients' dwellings during the epidemic of 1990-1992 was studied using the polymerase chain reaction (PCR). Amplification of miniexon-derived RNA genes and gpG3 mRNA was achieved in single Phlebotomus argentipes, P. papatasi, and Sergentomyia babu flies. The data suggest the possible involvement of multiple sandfly species in kala-azar transmission. PMID- 9347958 TI - Delayed migration of Plasmodium sporozoites from the mosquito bite site to the blood. AB - Studies were done on delivery of Plasmodium yoelii sporozoites by Anopheles stephensi into the skin of BALB/C mice. When infected mosquitoes fed on a portion of the ear, 81% of these positive control mice developed parasitemia. When the fed-upon site was excised immediately or 5 min postfeeding, a highly significant, smaller percentage of these experimental mice developed parasitemia. When the delay in removal of mosquito-bitten tissue was extended to 15 min, no significant difference was found between this group and positive control mice. These findings show that mosquito-injected sporozoites tend to remain at the bite site for at least 5 min after the mosquito bite. By approximately 15 min, the first wave of migrating sporozoites has left the bite site and moved into the general circulation. These findings have implications concerning possible host obstruction of sporozoite migration in skin by either anti-sporozoite antibodies or by a cutaneous hypersensitivity reaction to mosquito bite. PMID- 9347959 TI - Distinguishing Plasmodium falciparum treatment failures from reinfections by restrictions fragment length polymorphism and polymerase chain reaction genotyping. AB - The inability to distinguish recrudescent Plasmodium falciparum infections (treatment failures) from reinfections (new infections) is an important impediment to the evaluation of antimalarial treatment regimens. Ten paired primary and recrudescent isolates collected near the Thai-Cambodian border were analyzed by restriction fragment length polymorphism (RFLP) and by polymerase chain reaction (PCR) genotyping of the genes encoding the following proteins: circumsporozite (CS) protein, erythrocyte binding antigen (EBA)-175, ring infected erythrocyte surface antigen (RESA), merozoite surface protein-1 (MSP-1), and MSP-2. Both methods demonstrated that the fingerprint pattern of each recrudescent isolate was identical to or was contained within the pattern of the primary isolate. Each recrudescent isolate was unique when compared with the other nine primary isolates. Typing by PCR was more sensitive for the detection of multiclone infections and could be performed with small volumes of whole blood. The PCR genotyping could be a practical method for distinguishing a recrudescent from a new infection when treatment studies are conducted in areas with active malaria transmission. PMID- 9347960 TI - Enteropathogenic bacteria in the La Paz River of Bolivia. AB - Diarrheal diseases often result from ingestion of contaminated water or food. The population of La Paz, Bolivia is directly or indirectly exposed to the sewage contaminated La Paz River. We conducted a bacteriologic survey of the La Paz River to quantify the level of bacterial contamination, with particular reference to enteropathogens. A total bacterial count exceeding 10(6) colony-forming units (CFU)/ml, including lactose fermenting and nonfermenting, gram-negative bacilli of approximately 10(5) CFU/ml, respectively, were detected in river water samples collected near two densely populated areas. A total bacterial count of 10(5) CFU/ml was also detected at the most downstream area of the river near a sparsely populated area. At four sampling locations, several enteropathogens were detected, including five enterotoxigenic Escherichia coli (ETEC) (serotype O6, O15, and O159), two enteropathogenic E. coli (EPEC) (serotype O44), two enteroinvasive E. coli (EIEC) (serotype O29), and three Salmonella O4 group isolates. The heat-labile enterotoxin gene and the invasive toxin gene were detected in all ETEC and EIEC isolates by polymerase chain reaction analysis. Nine isolates of E. coli were found by the agar dilution method to be susceptible to ampicillin, kanamycin, nalidixic acid, tetracycline, and chloramphenicol, and ampicillin resistance was found in only two isolates of EIEC 7-4 (serotype O29) and EPEC 7-5 (serotype O44). Ampicillin resistance was coded on plasmids and transferred conjugatively to E. coli chi1037 at a frequency of 10(-5) CFU/donor by the broth mating method. Strains of Aeromonas caviae, which can cause diarrheal disease in infants, were detected in vegetables grown in fields irrigated by water from the La Paz River. The survival of nine isolates of E. coli in filtered river water was compared with that of laboratory strains (E. coli chi1037, W3110, and ATCC29577). The survival time of seven isolates, excluding two ampicillin-resistant isolates, was markedly longer than that of the laboratory strains. Our results show a high bacterial contamination of the La Paz river and suggest that such levels may contribute to the high incidence of diarrheal disease in the city of La Paz. PMID- 9347961 TI - Isolation, genetic diversity, and geographic distribution of Bayou virus (Bunyaviridae: hantavirus). AB - Bayou hantavirus, previously implicated in human hantavirus pulmonary syndrome in Louisiana, was isolated from a rice rat (Oryzomys palustris) captured in Georgia. The presence of antibody among rice rats captured throughout the southeastern United States and the extent of diversity among the genetic variants of Bayou viruses suggest that the rice rat is the most likely natural reservoir of the virus and that both virus and host have probably co-evolved for some years. PMID- 9347962 TI - A cluster of acute hepatitis E infection in United Nations Bangladeshi peacekeepers in Haiti. AB - In the fall of 1995, within a month of deployment to Haiti for peacekeeping duty, four Bangladeshi soldiers developed acute icteric hepatitis in rapid succession. Hepatitis E virus (HEV) was found to be the etiology by demonstrating HEV genomic sequences in serum samples by the polymerase chain reaction (PCR) and serologically by the detection of elevated IgM titers to HEV. No case had serologic evidence of acute hepatitis A or C infection. The soldiers had probably acquired their infection while living in a cantonment area outside Dhaka, Bangladesh for one month prior to deployment. Cloning and sequencing of amplified PCR products demonstrated a single strain suggestive of a common source of infection. Furthermore, high genomic identity with Asian strains of HEV and dissimilarity with the Mexican strain was demonstrated, verifying that the strain had indeed been imported. Human waste management from the Bangladesh camp in Haiti was strictly controlled and no secondary cases were observed. A convenience sample of 105 (12%) soldiers from the Bangladesh battalion (850 men) revealed anicteric or asymptomatic HEV infection in seven (7%) of 105. This report contains the first demonstration of acute hepatitis E in natives of Bangladesh and demonstrates the power of the PCR in the rapid diagnosis and epidemiologic analysis of HEV infection. More importantly, this cluster demonstrates the importation of an important infectious disease by multinational peacekeepers to a potentially susceptible host country. PMID- 9347963 TI - Short report: evidence of worldwide transmission of hepatitis G virus. AB - Hepatitis G virus (HGV) has been recently documented in the Americas, Europe, and Australia. Distinct risk populations from North Africa, South America, and Southeast Asia were screened for HGV, in addition to hepatitis B and C viruses. First time recognition of HGV is described from Egypt and Indonesia. Notable is the high proportion of HGV positive individuals among multiply transfused children, ranging from 24% of those sampled from Egypt to 32% in Indonesia. Also, data from Peru suggest the likely association of HGV infection with progressive liver disease. Hepatitis G virus should be considered a world-wide health concern. PMID- 9347964 TI - Emergence of raccoon rabies in Connecticut, 1991-1994: spatial and temporal characteristics of animal infection and human contact. AB - The North American raccoon rabies epizootic continues to expand, now affecting most of New England. In 1990, raccoons became the vertebrate most often reported rabid in the United States. Emergence of this zoonosis poses increasing, but poorly defined risks to humans. This study analyzed various demographic, environmental, and behavioral factors associated with animal infection and human exposure before and during the epizootic in Connecticut. Rabies virus infections among terrestrial vertebrates were analyzed from 1985 through 1994. From March 1991, when the first case was diagnosed, through December 1994, 2,522 of 13,147 animals tested were found positive for rabies viral antigen. Forty-seven percent of the raccoons tested were infected, representing 88.0% of all animals found positive. Domestic animals constituted only 1.7% of positive test results, but 40.6% of the tests performed. The epizootic wave of transmission advanced approximately 30 km/year. Most rabies-positive wild animals were taken from private properties, usually near houses. Possible human exposures involved 939 people on 556 occasions through direct contact (20.7%) or indirectly through another animal (79.3%). Of 3,239 domestic animals exposed to rabies-positive wild animals, 18.4% lacked vaccination. Rabies has become enzootic in Connecticut and risk to humans and animals persists. The public health burden is considerable, yet knowledge is lacking to develop sustainable prevention strategies. PMID- 9347965 TI - Determinants of infection with schistosomiasis haematobia using logistic regression. AB - A population-based stratified random sample of 10,039 inhabitants of rural communities in Minya Governorate, Egypt, were evaluated for risk factors for Schistosoma haematobium infection using multivariate analysis. Data were obtained by personal interview recording demographics, information on exposure to canal water, history of infection, and other risk factors for infection and examining urine samples for S. haematobium ova. Logistic regression analysis was used to adjust for confounders while assessing the role of each risk factor for infection. Using logistic regression allowed detection of several confounders and interactions which influenced other independent variables. Differences in exposure patterns to canal water among age and gender subgroups explained only a small portion of the variation in infection rates, thus favoring the alternative explanation: development of age-acquired immunity. The association of age with reduced prevalence of S. haematobium was the only relationship increasing (odds ratio [OR] = 2.95-4.30) with logistic regression. Male gender was a risk factor for infection but did not increase with logistic regression (OR = 2.33-2.03). The protective effects of education, only noted in schoolage children (OR = 0.59 0.51), were believed to be due to a school-based screening and treatment program. PMID- 9347966 TI - Spatial analysis of the distribution of LaCrosse encephalitis in Illinois, using a geographic information system and local and global spatial statistics. AB - The spatial and temporal distribution of LaCrosse encephalitis cases in Illinois was analyzed using a geographic information system (GIS) and spatial statistics. Case data were obtained from the Illinois Department of Public Health and mapped on the county, town, and address level. Human cases were concentrated in and around the city of Peoria in central Illinois. Local spatial statistics were used to identify hot spots where cases appear to be concentrated in the Peoria region. Several small towns surrounding the city of Peoria appeared as foci where cases were most common. Second-order spatial analysis of the case distribution was conducted on the address level. Cases were clustered within a range of 3.0 km in the city of Peoria. Since most cases appear to be associated with residential (peridomestic) exposure, and since several cases have been reported from neighboring addresses, transmission may be concentrated around specific sites (hardwood ravines, tire piles). The GIS and spatial analysis may be useful in identifying and targeting for intervention potential sites of enzootic transmission. PMID- 9347967 TI - Investigation of gray-headed flying foxes (Pteropus poliocephalus) (Megachiroptera: Pteropodidae) and mosquitoes in the ecology of Ross River virus in Australia. AB - Entomologic and virologic factors were investigated to determine whether gray headed flying foxes (Pteropus poliocephalus) from Indooroopilly Island, Brisbane, Australia could be vertebrate hosts of Ross River (RR) virus. Aedes funereus was the most abundant mosquito species with 6,300-38,700 females per light trap night in the flying fox camp containing gray-headed, black (P. alecto), and little red (P. scapulatus) flying foxes. Sixteen Ae. funereus blood meals from this collection were analyzed by hemoglobin electrophoresis and were found to be from P. alecto. From pledget feeding with RR virus, the infectious dose required to infect 50% of wild caught Ae. funereus was log10 4.2 50% tissue culture infectious doses per mosquito, with a transmission rate to mice of 17% at 9-10 days post infection. Experimental infection of 10 juvenile P. poliocephalus produced viremias of low titer in five animals, with a duration of 1-4 days and a mean of two days. Three percent of colonized Ae. vigilax that fed on the 10 animals during this period became infected. One of the five viremic flying foxes and two of the five aviremic animals produced a detectable immune response by either neutralization or hemagglutination-inhibition tests. Based on the low to moderate vector competence of Ae. funereus for RR virus, and evidence that P. poliocephalus is a poor vertebrate host of RR virus, it is unlikely that RR virus transmission would be maintained between these two species, but it could be maintained by other more competent vector/host pairs. PMID- 9347968 TI - Clearance of Wuchereria bancrofti antigen after treatment with diethylcarbamazine or ivermectin. AB - The present study was undertaken to assess the relationship between microfilarial clearance and clearance of circulating filarial antigen from the blood of Wuchereria bancrofti-infected persons following chemotherapy with either diethylcarbamazine or ivermectin. Patients received either 12 weekly doses of 6 mg/kg of diethylcarbamazine (DEC), a single dose of 6 mg/kg of DEC, a single dose of 420 microg/kg of ivermectin, or 20 microg/kg of ivermectin, followed by 6 mg/kg of DEC five days later. Microfilarial clearance was marked in all groups, but was significantly less in the single-dose DEC. In contrast, as monitored by the Og4C3 monoclonal anitbody assay, clearance of circulating filarial antigen was highly variable, not only between groups but within each group. As a result, there were few statistically significant differences in antigen clearance between groups. In no instance did the antigen level fall to zero, even in individuals that remained microfilaria negative during two or three years of follow-up. These results suggest that living adult worms persist in almost all persons treated with DEC and/or ivermectin. PMID- 9347969 TI - Factors affecting high and low human IgE responses to schistosome worm antigens in an area of Brazil endemic for Schistosoma mansoni and hookworm. AB - We have previously examined the antibody isotype responses to schistosome worm and egg antigens in human populations living in areas of Kenya and the Philippines endemic for Schistosoma mansoni and S. japonicum, respectively. Here, we have analyzed antibody isotype responses to S. mansoni adult worm (AW) antigen and soluble egg antigen (SEA) in more than 500 Brazilian individuals, and found similar relationships with age and sex as in the Kenyan and Filipino populations. Isotype responses to AW antigen broadly increased with age whereas isotype responses to SEA decreased, and a higher proportion of males than females had detectable IgE against AW antigen. Most isotype responses to AW antigen and SEA correlated positively with intensity of infection with S. mansoni except AW antigen-specific IgG2, which correlated negatively. The overall prevalence of infection with S. mansoni in this area was relatively low at only 39.5%; hookworm prevalence was higher at 57.4%. The majority of those infected with S. mansoni were also infected with hookworm (76%). Those individuals with high IgE responses to AW antigen were matched for sex, age, and total IgG to AW antigen as closely as possible with individuals with low levels of AW antigen-specific IgE. The two groups were compared for factors potentially influential in IgE production. No difference was found between the high and low IgE responders for 1) intensity or prevalence of infection with S. mansoni, 2) relative exposure to S. mansoni, 3) number of previous treatments for schistosomiasis, or 4) prevalence of infection with hookworm, but differences were found in other isotype responses to S. mansoni. The high IgE responders had higher IgA and IgG4 against both AW antigen and SEA but lower IgG3 responses to AW antigen than the low IgE responders. The IgE responses to three S. mansoni antigens (paramyosin, Sm22.6, and a 12-kD AW antigen band) were detected in individuals with IgE against AW antigen only. PMID- 9347970 TI - Measurement of Plasmodium falciparum growth rates in vivo: a test of malaria vaccines. AB - Several prototype vaccines against the asexual blood stage of malaria are undergoing preclinical and phase I testing. Although these vaccines have been chosen for their ability to elicit an anti-parasite response, no practical and sensitive clinical trial procedure has been available for measuring their impact on parasite growth. We describe a system that allows parasite growth rates to be measured in volunteers through the incubation period. Two necessary elements of this system are developed: suitable blood-stage Plasmodium falciparum inocula, and a highly sensitive and quantitative assay to measure parasite growth during the incubation period. We infected five nonimmune volunteers with an inoculum as small as 300 parasites and demonstrated that the resultant in vivo asexual parasite growth rates were reproducible at 12-15-fold per cycle. The system allowed the infection to be followed for eight days before treatment without symptoms developing. These findings suggest that it is feasible to directly measure the anti-parasite efficacy of a prototype malaria vaccine in human volunteers without subjecting them to the risk of disease. PMID- 9347971 TI - In times of change learners inherit the earth. The 1997 presidential address. AB - Neurosurgery, as traditionally practiced, is changing. Eric Hoffer observed, "In times of change learners inherit the earth; while the learned find themselves beautifully equipped to deal with a world that no longer exists." Historically, learners have possessed certain attributes. They anticipate rather than react to change. They become essential facilitators within their altered environment. They are effective advocates for principled behavior and ethical practices. They skillfully communicate the new and technically arcane with a common touch. Learners also understand the lessons of history. The potential of process technology, illustrated by Morton's 1846 application of already well-understood science, in essence, to transform the profession of surgery by making painless operations possible, is one example. Voluntary professional associations such as organized neurosurgery can also learn from the contrasting experiences of the Donner Party and the Utah Pioneers as they made their way down the same canyon of the Wasatch mountain range during the summers of 1846 and 1847, respectively. Group dynamics during those two episodes differed strikingly. It was the vanguard party of Utah Pioneers who exemplified the characters of learners. They acted with unity, common purpose, and careful preparation designed to benefit those who followed them. Years from now, how will the current individual and institutional behavior of neurosurgeons and their associations be judged? An interval assessment is suggested at the April, 2012 American Association of Neurological Surgeons Annual Scientific Meeting. From that perspective, will it be determined that the neurosurgical discipline acted presciently, as would those Hoffer describes as learners? Will neurosurgery be said to have understood and thereby profited from the lessons of history? These are pertinent questions. The answers, considered as a proposed progress report 15 short years from now, could accurately predict the scientific and professional position that neurological surgery will occupy in the future. Deserved long-term success based on the effective, far-sighted behavior exhibited by learners is my confident expectation. PMID- 9347972 TI - Nonsurgical, nonorthotic treatment of occipital plagiocephaly: what is the natural history of the misshapen neonatal head? AB - Management of neonates with nonsynostotic occipital plagiocephaly has been controversial, and there has been a lack of uniformity concerning its treatment. Patients with nonsynostotic occipital plagiocephaly have been treated surgically or with cranial remodeling orthotic devices and have shown improvement in asymmetry. The cost of orthotic treatment has risen, and its validity has been contested by many third-party insurance payers. The effectiveness of orthotic treatment has not been adequately compared to the natural history of nonsynostotic occipital plagiocephaly. A nonsurgical, nonorthotic treatment study was initiated in June 1995 at Phoenix Children's Hospital. All new patients referred with a diagnosis of nonsynostotic occipital plagiocephaly were categorized into two groups: those with mild-to-moderate asymmetry and those with moderate-to-severe asymmetry. Categories were determined by cephalic measurements. The patients with moderate-to-severe asymmetry were offered orthotic treatment with a cranial remodeling band. Those patients with mild-to moderate asymmetry were treated with physiotherapy, repositioning of the head, and repeated notation of cephalic measurements without orthotic devices or surgery. Seventy-two neonates, seen consecutively, with mild-to-moderate, nonsynostotic occipital plagiocephaly were evaluated by noting cephalic measurements. The parents of six of these patients elected treatment with a cranial remodeling band and results in these patients were excluded from our data. The remaining 66, treated without orthotic devices, showed improvement in average cranial vault asymmetry (CVA) from 9.2 to 4.7 mm over an average treatment period of 4.5 months that commenced when the average age of the patient was 6.4 months. A comparison of the present data with data published in 1994 for neonates treated with a headband indicates that neonates with mild-to-moderate asymmetry who are treated aggressively with physiotherapy and repositioning have similar improvement in CVA. PMID- 9347974 TI - Transpalatal approach for the extracranial surgical repair of transsphenoidal cephaloceles in children. AB - The surgical treatment of transsphenoidal cephaloceles in children is controversial. Reduction and repair via a transcranial approach are associated with high postoperative rates of morbidity, mortality, and hypothalamic dysfunction. In this study, four patients, aged 3 to 35 months at surgery, underwent successful transpalatal repair of two encephaloceles and two meningoceles. Two patients presented with nasal obstruction in infancy, one presented with unexplained meningitis, and in one patient the lesion was found incidentally during evaluation for seizures. Two children had median cleft face syndrome, another had an associated Arnold-Chiari type I malformation, and the fourth had no other cranial abnormalities. All patients underwent preoperative evaluation including magnetic resonance (MR) imaging. Auditory, ophthalmological, genetic, endocrinological, or other evaluation was undertaken as indicated. Lesions were approached through the median raphe of the hard and soft palates. All cephaloceles were easily visualized and dissected after division of the nasal palatal mucosa. The dural sac and its contents were reduced by surface coagulation after division and dissection of the overlying mucosa. Once reduced, the bone defect was obliterated in three of four patients. The dura was not opened and anomalous neural elements were not resected. At follow-up evaluation, all patients demonstrated resolution of preoperative symptoms without evidence of infection or lasting morbidity. Follow-up MR imaging showed reduction in all cases. The authors conclude that this transpalatal approach is safe and reliable for the treatment of transsphenoidal cephaloceles in young children. PMID- 9347973 TI - Upper cervical spine fusion in the pediatric population. AB - The outcomes of 25 pediatric patients who underwent upper cervical or occipitocervical fusion at the authors' institution since 1983 were reviewed. At a mean age of 9 years, the patients presented with spinal instability that was associated with os odontoideum in 11 cases, rotatory subluxation in five cases, odontoid fracture in two cases, atlantooccipital dislocation in two cases, and congenital atlantoaxial instability in five patients, four of whom had Down's syndrome (trisomy 21). Ten children had abnormal findings on neurological examination preoperatively; however, nine experienced improvement or resolution of deficits as of their latest follow-up evaluation (mean 17 months). Fusion was achieved with the first operation in 21 of 25 patients; eventually it was attained in all but one. Four patients exhibited persistent spinal instability after an initial procedure. This was caused by erosion of a multistranded cable through the intact arch of C-2 in two cases, by pin site infection necessitating early halo removal in one case, and by slippage in a halo following a Gallie procedure, which was revised with a Brooks fusion in one case. This series, the largest yet published, shows that with appropriate surgical management, posterior upper cervical fusion in the pediatric population is highly successful. Careful attention to halo pin site care and caution in using multistranded cable in young patients may improve results. PMID- 9347975 TI - Increased risk of distal ventriculoperitoneal shunt obstruction associated with slit valves or distal slits in the peritoneal catheter. AB - The authors describe a relationship between the presence of distal shunt catheter side-wall slits and distal catheter obstruction in a single-surgeon series of ventriculoperitoneal (VP) shunt revisions. Between 1985 and 1996, 168 operations for VP shunt revision were performed by the senior author (J.W.C.) in 71 patients. Indications for shunt revision included obstruction in 140 operations; overdrainage or underdrainage requiring a change of valve in 17 operations; inadequate length of distal shunt tubing resulting in the distal end no longer reaching the peritoneum in five operations; the ventricular catheter in the wrong ventricle or space, requiring repositioning in five operations; and a disconnected or broken shunt in one operation. Of the 140 instances of shunt obstruction, the blockage occurred at the ventricular end in 108 instances (77.1%), the peritoneal end in 17 (12.1%), the ventricular and the peritoneal end in 14 (10%), and in the valve mechanism (not including distal slit valves) in one (0.8%). Thus, the peritoneal end was obstructed in 31 (22.1%) of 140 cases of shunt malfunction. In every case in which the peritoneal end was obstructed, some form of distal slit was found: either a distal slit valve in an otherwise closed catheter or slits in the side of an open catheter. No instances were found of distal peritoneal catheter obstruction when the peritoneal catheter was a simple open-ended tube with no accompanying side slits (0 of 55). It is concluded that side slits in the distal peritoneal catheters of VP shunts are associated with a greater incidence of distal shunt obstruction. PMID- 9347976 TI - Dutch normal-pressure hydrocephalus study: prediction of outcome after shunting by resistance to outflow of cerebrospinal fluid. AB - The authors examined whether measurement of resistance to outflow of cerebrospinal fluid (Rcsf) predicts outcome after shunting for patients with normal-pressure hydrocephalus (NPH). In four centers 101 patients (most of whom had idiopathic NPH) who fulfilled strict entry criteria underwent shunt placement irrespective of their level of Rcsf obtained by lumbar constant flow infusion. Gait disturbance and dementia were quantified by using an NPH scale and the patient's level of disability was assessed by using the modified Rankin scale (mRS). In addition the Modified Mini-Mental State Examination was performed. Patients were assessed prior to and 1, 3, 6, 9, and 12 months after surgery. Primary outcome measures were based on differences between the preoperative and last NPH scale scores and mRS grades. Improvement was defined as a change measuring at least 15% in the NPH scale score and at least one mRS grade. Intention-to-treat analysis of all patients at 1 year yielded improvement for 57% in NPH scale score and 59% in mRS grade. Efficacy analysis, excluding serious events and deaths that were unrelated to NPH, was performed for 95 patients. Improvement rose to 76% in NPH scale score and 69% in mRS grade. Six cut-off levels of Rcsf were related to improvement in NPH scale score using two-by-two tables. Positive predictive values were approximately 80% for an Rcsf of 10, 12, or 15 mm Hg/ml/minute, 92% for an Rcsf of 18 mm Hg/ml/minute, and 100% for an Rcsf of 24 mm Hg/ml/minute. Negative predictive values were low. More important was the highest likelihood ratio of 3.5 for an Rcsf of 18 mm Hg/ml/minute. Extensive comorbidity was a major prognostic factor. Measurement of Rcsf reliably predicts outcome if the limit for shunting is raised to 18 mm Hg/ml/minute. At lower Rcsf values the decision depends mainly on the extent to which clinical and computerized tomography findings are typical of NPH. PMID- 9347978 TI - Pallidal stimulation: an alternative to pallidotomy? AB - A resurgence of interest in the surgical treatment of Parkinson's disease (PD) came with the rediscovery of posteroventral pallidotomy by Laitinen in 1985. Laitinen's procedure improved most symptoms in drug-resistant PD, which engendered wide interest in the neurosurgical community. Another lesioning procedure, ventrolateral thalamotomy, has become a powerful alternative to stimulate the nucleus ventralis intermedius, producing high long-term success rates and low morbidity rates. Pallidal stimulation has not met with the same success. According to the literature pallidotomy improves the "on" symptoms of PD, such as dyskinesias, as well as the "off" symptoms, such as rigidity, bradykinesia, and on-off fluctuations. Pallidal stimulation improves bradykinesia and rigidity to a minor extent; however, its strength seems to be in improving levodopa-induced dyskinesias. Stimulation often produces an improvement in the hyper- or dyskinetic upper limbs, but increases the "freezing" phenomenon in the lower limbs at the same time. Considering the small increase in the patient's independence, the high costs of bilateral implants, and the difficulty most patients experience in handling the devices, the question arises as to whether bilateral pallidal stimulation is a real alternative to pallidotomy. PMID- 9347977 TI - Complications associated with intraventricular chemotherapy in patients with leptomeningeal metastases. AB - The authors studied complications associated with intraventricular chemotherapy in patients with leptomeningeal metastases (LM). One hundred twenty consecutive patients with LM (71 females and 49 males) ranging in age from 10 to 72 years (median 42 years) were treated with involved-field radiotherapy and intraventricular chemotherapy using an Ommaya reservoir and intraventricular catheter system. The diagnosis of LM was determined by a combination of clinical presentation (114 patients); cerebrospinal fluid cytological studies (100); or neuroradiographic studies (42). Systemic tumor histological findings included breast (34 patients); non-Hodgkin's lymphoma (22); melanoma (16); primitive neuroectodermal tumors including medulloblastoma (10); glial neoplasms, leukemia, small cell lung, nonsmall cell lung, and colon (six each); prostate and kidney (three each); and gastric cancers (two). Sixteen patients, all with non-Hodgkin's lymphoma, also had acquired immune deficiency syndrome. Patients received one to four (median two) chemotherapeutic drugs and underwent a total of 1110 cycles of intraventricular chemotherapy (median 10). Intraventricular chemotherapy administration and diagnostic Ommaya reservoir punctures totaled 4400, with a median of 46 per patient. Complications included aseptic/chemical meningitis (52 patients); myelosuppression due to intraventricular chemotherapy (21); catheter related infections (nine); unidirectional catheter obstruction (six); intraventricular catheter malpositioning (two); Ommaya reservoir exposure (two); leukoencephalopathy (two); and chemotherapy-related myelopathy (one). There were no treatment-related deaths; however, seven patients (6%) required additional surgery for either catheter repositioning (two) or reservoir removal (five). Seven patients with catheter-related infections were treated successfully with intraventricular and systemic antibiotic drugs, thereby preserving the Ommaya system. The authors conclude that Ommaya reservoirs are convenient and pharmacologically rational systems for administering intraventricular chemotherapy. Overall, serious complications requiring surgery are infrequent (6%) and most often secondary to catheter infections, Ommaya reservoir exposure, or initial catheter malpositioning. In the majority of instances, catheter infections may be managed medically, as may the most common complications of intraventricular chemotherapy including aseptic meningitis (43% of patients) and myelosuppression (18%). PMID- 9347979 TI - The transcallosal-transforaminal approach to the third ventricle with regard to the venous variations in this region. AB - Surgical approaches to lesions located in the anterior and middle portions of the third ventricle are challenging, even for experienced neurosurgeons. Various exposures involving the foramen of Monro, the choroidal fissure, the fornices, and the lamina terminalis have been advocated in numerous publications. The authors conducted a microsurgical anatomical study in 20 cadaveric brain specimens (40 hemispheres) to identify an exposure of the third ventricle that would avoid compromising vital structures. An investigation of the variations in the subependymal veins of the lateral ventricle in the region of the foramen of Monro was performed, as these structures are intimately associated with the surgical exposure of the third ventricle. In 16 (80%) of the brain specimens studied, 19 (47.5%) of the hemispheres displayed a posterior location of the anterior septal vein-internal cerebral vein (ASV-ICV) junction, 3 to 13 mm (average 6 mm) beyond the foramen of Monro within the velum interpositum, not adjacent to the posterior margin of the foramen of Monro (the classic description). Based on this finding, the authors advocate opening the choroidal fissure as far as the ASV-ICV junction to enlarge the foramen of Monro posteriorly. This technique achieves adequate access to the anterior and middle portions of the third ventricle without causing injury to vital neural or vascular structures. The high incidence of posteriorly located ASV-ICV junctions is a significant factor influencing the successful course of surgery. Precise planning of the surgical approach is possible, because the location of the junction is revealed on preoperative neuroradiological studies, in particular on magnetic resonance venography. It can therefore be determined in advance which foramen of Monro qualifies for posterior enlargement to gain the widest possible access to the third ventricle. This technique was applied in three patients with a third ventricular tumor, and knowledge of the venous variations in this region was an important resource in guiding the operative exposure. PMID- 9347980 TI - Cathepsin B and middle cerebral artery occlusion in the rat. AB - Lysosomal proteases, although tightly regulated under physiological conditions, are known to contribute to cell injury after various forms of tissue ischemia have occurred. Because cathepsin B is a prominent lysosomal protease found in brain parenchyma, the authors hypothesized that it may contribute to neuronal cell death after focal cerebral ischemia. The authors measured the expression and spatial distribution of cathepsin B within the ischemic brain in 43 animals by means of immunohistochemical analysis in a rat model of transient middle cerebral artery (MCA) occlusion. Cathepsin B activity was also measured within specific ischemic brain regions by using an in vitro assay (22 animals). In addition, the authors tested the therapeutic effect of preischemic intraventricular administration of stefin A, a cysteine protease inhibitor, on the volume of cerebral infarction after transient MCA occlusion (15 animals). Increased cathepsin B immunoreactivity was detected exclusively within the ischemic neurons after 2 hours of reperfusion following a 2-hour MCA occlusion. Cathepsin B immunolocalization in the ischemic region decreased by 24 hours of reperfusion, but then increased by 48 hours of reperfusion because the infarct was infiltrated by inflammatory cells. Increased immunolocalization of cathepsin B in the inflammatory cells located in the necrotic infarct core continued through 7 days of reperfusion. Cathepsin B enzymatic activity was significantly increased in the ischemic tissue at 2, 8, and 48 hours, but not at 24 hours of reperfusion after 2 hours of MCA occlusion. Continuous intraventricular infusion of stefin A, before 2 hours of MCA occlusion (15 animals), significantly reduced infarct volume compared with control animals (12 animals): the percentage of hemispheric infarct volume was 20+/-3.9 compared with 33+/-3.5 (standard error of the mean; p = 0.025). These data indicate that neuronal cathepsin B undergoes increased expression and activation within 2 hours of reperfusion after a 2-hour MCA occlusion and may be a mechanism contributing to neuronal cell death. Intraventricular infusion of stefin A, an inhibitor of cathepsin B, significantly reduces cerebral infarct volume in rats. PMID- 9347981 TI - Estrogens may reduce mortality and ischemic damage caused by middle cerebral artery occlusion in the female rat. AB - The present study was undertaken to determine if estrogens protect female rats from the neurodegenerative effects of middle cerebral artery (MCA) occlusion. The rats were ovariectomized and 7 or 8 days later various estrogen preparations were administered before or after MCA occlusion. Pretreatment with 17beta-estradiol (17beta-E2) or a brain-targeted 17beta-E2 chemical delivery system (CDS) decreased mortality from 65% in ovariectomized rats to 22% in 17beta-E2-treated and 16% in 17beta-E2 CDS-treated rats. This marked reduction in mortality was accompanied by a reduction in the ischemic area of the brain from 25.6+/-5.7% in the ovariectomized rats to 9.8+/-4% and 9.1+/-4.2% in the 17beta-E2-implanted and the 17beta-E2 CDS-treated rats, respectively. Similarly, pretreatment with the presumed inactive estrogen, 17alpha-estradiol, reduced mortality from 36 to 0% and reduced the ischemic area by 55 to 81%. When administered 40 or 90 minutes after MCA occlusion, 17beta-E2 CDS reduced the area of ischemia by 45 to 90% or 31%, respectively. In summary, the present study provides the first evidence that estrogens exert neuroprotective effects in an animal model of ischemia and suggests that estrogens may be a useful therapy to protect neurons against the neurodegenerative effects of stroke. PMID- 9347983 TI - Noninvasive optical imaging of the subarachnoid space and cerebrospinal fluid pathways based on near-infrared fluorescence. AB - The authors have developed a noninvasive optical method to image the subarachnoid space and cerebrospinal fluid pathways in vivo based on the near-infrared fluorescence of indocyanine green (ICG). The ICG was bound to purified lipoproteins (ICG-lipoprotein) and injected into the subarachnoid space of neonatal and adult rats. The ICG fluorescence was detected by a cooled charge coupled device camera. After injection of ICG-lipoprotein into the cerebral subarachnoid space of the neonatal rat, ICG fluorescence was clearly detected at the injection site through the skull and skin. The ICG fluorescence was observed in the cerebellum and the lumbar spinal cord 1 and 8 hours postinjection, respectively. After injection of ICG-lipoprotein into the lumbar spinal subarachnoid space of an adult rat, ICG fluorescence was observed from the injection site to the thoracic levels along the spinal subarachnoid space. In addition, with the rat's head tilted downward, ICG fluorescence had extended to the cerebral subarachnoid space by 1 hour postinjection. The ICG fluorescence imaging of the cerebral subarachnoid space demonstrated an increase in fluorescence intensity around the lambdoid suture and the forebrain. On dissection of the rat brain the former location was identified as the supracerebellar cistern and the latter as the olfactory cistern. The results of this study are the first to demonstrate that an optical technique is applicable to imaging of the subarachnoid space and cerebrospinal fluid pathways in vivo. In addition, ICG-lipoprotein provides a sensitive optical tracer for imaging extravascular biological structures. Finally, ICG fluorescence imaging does not require an intricate imaging system because ICG is localized near the surface of the body. PMID- 9347982 TI - Attenuation of brain injury and reduction of neuron-specific enolase by nicardipine in systemic circulation following focal ischemia and reperfusion in a rat model. AB - A reversible middle cerebral artery occlusion was performed in rats to determine whether nicardipine, a dihydropyridine voltage-sensitive Ca++ channel (VSCC) antagonist, exerts neuroprotective effects when administered 10 minutes following an ischemic insult, and if it does, whether this is due to its vasodilatory action and effect on cerebral blood flow (CBF) or to direct blockade of Ca++ entry into ischemic brain cells. An increase in the intracellular calcium, [Ca++]i, plays a major role in neuronal injury during cerebral ischemia. Although a large amount of Ca++ enters neurons through the VSCC during ischemia, inconsistent neuroprotective effects have been reported with the antagonists of the VSCC. An intraperitoneal injection of nicardipine (1.2 mg/kg) was administered to rats 10 minutes after the onset of ischemia, and 8, 16, and 24 hours after occlusion. Cortical CBF was determined by laser-Doppler flowmetry. Neurological and neuropathological examinations were performed after 72 hours. Neuron-specific enolase, a specific marker for the incidence of neuronal injury, was measured in plasma. The CBF and other physiological parameters were not affected by nicardipine during occlusion or reperfusion. However, nicardipine treatment significantly improved motor neurological outcome by 29%, and the infarction and edema volume in the pallium as well as the edema volume in the striatum were significantly reduced by 27%, 37%, and 52%, respectively. Nicardipine also reduced the neuron-specific enolase plasma levels by 50%, 42%, and 59% at 24, 48, and 72 hours after the occlusion, respectively. It is concluded that nicardipine may attenuate focal ischemic brain injury by exerting direct neuroprotective and antiedematous effects that do not depend on CBF. PMID- 9347984 TI - Reversal and prevention of cerebral vasospasm by intracarotid infusions of nitric oxide donors in a primate model of subarachnoid hemorrhage. AB - Decreased endothelium-derived relaxing factor, nitric oxide (NO), in the arterial wall has been hypothesized to be a potential cause of cerebral vasospasm following subarachnoid hemorrhage (SAH). The authors sought to determine whether intracarotid infusions of newly developed NO-donating compounds (NONOates) could reverse vasospasm or prevent the occurrence of cerebral vasospasm in a primate model of SAH. Twenty-one cynomolgus monkeys were studied in two experimental settings. In an acute infusion experiment, saline or NONOate was infused intracarotidly in four normal monkeys and in four monkeys after onset of SAH. During the infusions regional cerebral blood flow (rCBF) was measured in eight animals and CBF velocity in two. In a chronic infusion experiment, saline (four animals) or NONOate (diethylamine-NO [three animals] or proli-NO [six animals]) was infused intracarotidly in monkeys for 7 days after SAH. In acute infusion experiments, 3-minute intracarotid diethylamine-NO infusions reversed arteriographically confirmed vasospasm of the right middle cerebral artery (MCA) (as viewed on anteroposterior projection, the decrease in area was 8.4+/-4.3% in the treatment group compared with 35+/-12% in the control group; p < 0.004), increased rCBF by 31+/-1.9% (p < 0.002), and decreased the mean systolic CBF velocity in the right MCA. In a long-term infusion experiment, the area of the right MCA in control animals decreased by 63+/-5%. In animals undergoing a 7-day continuous glucantime-NO intracarotid infusion, the area of the right MCA decreased by 15+/-6.2%, and in animals undergoing a 7-day proli-NO infusion, the area of the right MCA decreased by 11+/-2.9% (p < 0.05). The mean arterial blood pressure decreased in the glucantime-NO group from 75+/-12 mm Hg (during saline infusion) to 57+/-10 mm Hg (during glucantime-NO infusion; p < 0.05), but it was unchanged in animals undergoing proli-NO infusion (76+/-12 mm Hg vs. 78+/-12 mm Hg). Results of these experiments show that cerebral vasospasm is both reversed and completely prevented by NO replacement. However, only the use of regional infusion of the NONOate with an extremely short half-life avoided a concomitant decrease in arterial blood pressure, which could produce cerebral ischemia in patients with impaired autoregulation of CBF after the rupture of an intracranial aneurysm. PMID- 9347985 TI - Generation of the catalytic fragment of protein kinase C alpha in spastic canine basilar artery. AB - In previous studies of topical application of calphostin C, a specific inhibitor of the regulatory domain of protein kinase C (PKC), and calpeptin, a selective inhibitor of calpain, to spastic canine basilar artery (BA) researchers have suggested that the catalytic fragment of PKC (known as PKM) is probably formed by a limited proteolysis of continuously activated mu-calpain, but there has been no direct evidence for PKM formation in vasospasm. The present immunoblot study with anti-PKCalpha antibody shows a significant decrease in cytosolic 80-kD PKCalpha and a concomitantly significant increase in membrane PKCalpha in the spastic canine BA. In addition, an immunoblot study in which cleavage site-directed antibodies were used demonstrated a significant increase in immunoreactive 45-kD PKM. The changes in membrane PKCalpha and PKM were enhanced with the lapse of time after subarachnoid hemorrhage. The cleavage site-directed antibodies distinguish the proteolyzed from the unproteolyzed forms of PKC for in situ analyses of enzyme regulation mediated by proteolysis. The data indicate that PKCalpha in spastic canine BA is translocated to the cell membrane, where PKCalpha is rapidly cleaved into PKM as a result of proteolysis of the isozyme by mu-calpain but not by m-calpain. The authors hypothesize that mu-calpain is continuously activated in spastic canine BA and produces PKM by limited proteolysis of PKCalpha. PMID- 9347986 TI - Completely dislocated hangman's fracture with a locked C2-3 facet. Case report. AB - The authors report a rare case of a hangman's fracture involving complete dislocation of C-2 onto C-3, accompanied by a C2-3 locked facet and asymptomatic bilateral vertebral artery injuries. The patient, a 25-year-old man who sustained a neck injury in an industrial accident, presented with a mild central spinal cord syndrome. His initial lateral cervical radiograph showed complete anterior dislocation of the C-2 body onto C-3, bilateral neural arch fractures, and a unilateral locked facet. The mechanism was likely flexion and compression. The grossly unstable spine and the locked facet were treated by posterior decompression, reduction, and C1-3 fixation. The patient recovered in several days and is without neurological deficit. PMID- 9347987 TI - Paraspinal calcinosis associated with progressive systemic sclerosis. Case report. AB - The authors describe a case of paraspinal calcinosis in a 65-year-old woman with progressive systemic sclerosis. Although calcinosis occurs in up to 27% of cases of progressive systemic sclerosis, symptomatic paraspinal calcinosis is extremely rare. In the case reported here, multiple cervical facet joints were compromised by progressive calcinosis, leading to glacial spinal instability. Internal fixation was indicated to correct the instability and decompress the spinal canal. Medical therapy was instituted to arrest or reverse the ongoing calcinosis. PMID- 9347988 TI - Pheochromocytoma and multiple intracranial aneurysms: is it a coincidence? Case report. AB - The authors present a case of multiple intracranial aneurysms associated with a pheochromocytoma. The aneurysms were successfully clipped, and a suprarenal tumor located on the left side was totally removed. To the authors' knowledge this is the fourth reported case of these combined entities in the literature. The authors speculate on the possible etiopathogenesis of the relationship between intracranial aneurysms and attacks of hypertension caused by the presence of neoplasms that discharge acute and irregular levels of catecholamines, especially pheochromocytomas. Perioperative management designed to avoid undesired complications in this rare association is also discussed. PMID- 9347989 TI - Microsurgical excision of a primary isolated hypothalamic eosinophilic granuloma. Case report. AB - Solitary focal eosinophilic granuloma (EG) is one element in the spectrum of diseases associated with Langerhans' cell histiocytosis (LCH). This report documents the occurrence of a primary isolated hypothalamic EG in a man who presented with diabetes insipidus and panhypopituitarism. His treatment consisted of complete microsurgical excision of the lesion. After a 13-month follow-up period, no residual tumor was evident on magnetic resonance imaging and no other lesions were present in peripheral tissues. This case is unique in several respects: 1) it is the third documented case of a primary isolated hypothalamic LCH granuloma diagnosed in a living patient; 2) it is the only known example of complete microsurgical excision of such a lesion in the hypothalamic region; and 3) it demonstrates the efficacy of direct surgery in this scenario, as compared with other treatment modalities such as biopsy and irradiation, suggesting that complete surgical excision may represent the treatment of choice for isolated intracerebral LCH granulomas, being curative in most instances. Also, the literature is reviewed for information about the diagnosis and treatment of this particular type of unifocal brain lesion. PMID- 9347990 TI - Massive osteolysis of the skull and upper cervical spine. Case report and review of the literature. AB - Massive osteolysis is a type of idiopathic osteolysis in which there is spontaneous onset of bone resorption. Almost any bone in the body can be affected. The authors present the case of a 62-year-old man diagnosed with massive osteolysis of the occipital bone and the upper two cervical vertebrae. Despite extensive pneumocephalus, no neurological sign or spinal instability was evident. In this case 4000 cGy of radiation in 200-cGy fractions was administered to the diseased area while the patient was kept in a Miami-J collar. At the 2 year follow-up examination, arrest of the disease process and new bone formation was evident on radiographic studies. PMID- 9347991 TI - Total sacrectomy and Galveston L-rod reconstruction for malignant neoplasms. Technical note. AB - Although radical resection is the best treatment for malignant sacral tumors, total sacrectomy for such tumors has been performed in only a few instances. Total sacral resection requires reconstruction of the pelvic ring plus establishment of a bilateral union between the lumbar spine and iliac bone. This technique is illustrated in two patients harboring large, painful, sacral giant cell tumors that were unresponsive to prior treatment. These patients were treated with complete en bloc resection of the sacrum and complex iliolumbar reconstruction/stabilization and fusion. Surgery was performed in two stages, the first consisting of a midline celiotomy, dissection of visceral/neural structures, and ligation of internal iliac vessels, followed by an anterior L5-S1 discectomy. The second stage consisted of mobilization of an inferiorly based myocutaneous rectus abdominis pedicle flap for wound closure, followed by an L-5 laminectomy, bilateral L-5 foraminotomy, ligation of the thecal sac, division of sacral nerve roots, and transection of the ilia lateral to the tumor and sacroiliac joints. Placement of the instrumentation required segmental fixation of the lumbar spine from L-3 down by means of pedicle screws and the establishment of a bilateral liaison between the lumbar spine and the ilia by using the Galveston L-rod technique. The pelvic ring was then reestablished by means of a threaded rod connecting left and right ilia. Both autologous (posterior iliac crest) and allograft bone were used for fusion, and a tibial allograft strut was placed between the remaining ilia. The patients were immobilized for 8 weeks postoperatively and underwent progressive rehabilitation. At the 1-year follow-up review, one patient could walk unassisted, and the other ambulated independently using a cane. Both patients controlled bowel function satisfactorily with laxatives and diet and could maintain continence but required self-catheterization for bladder emptying. The authors conclude that in selected patients, total sacrectomy represents an acceptable surgical procedure that can offer not only effective local pain control, but also a potential cure, while preserving satisfactory ambulatory capacity and neurological function. PMID- 9347992 TI - Aneurysm clips: magnetic quantification and magnetic resonance imaging safety. Technical note. AB - Knowledge of the magnetic properties of cerebral aneurysm clips in patients undergoing magnetic resonance (MR) imaging is imperative. The authors quantified in electromagnetic units the magnetic properties of 13 different types of aneurysm clips by using a vibrating sample magnetometer. Their results showed that the magnetic moment of these clips ranged from 0.15 EMU/g to as high as 152.7 EMU/g. Based on these results and tests of the movement of the clips during MR imaging, they conclude that aneurysm clips with a magnetic moment less than 1 EMU/g may be safely used during MR imaging. The quantification of magnetic properties into electromagnetic units by using a vibrating sample magnetometer is a reliable method applicable to any testing field gradient. This method can be used as a standard to measure and label the magnetic properties of aneurysm clips. PMID- 9347993 TI - Reversed-flow saphenous vein grafts for cerebral revascularization. Technical note. AB - The authors sought to create a saphenous vein interposition graft to be used in cerebral bypass procedures that would be more physiologically appropriate than standard vein grafts and would provide a better match between the graft and recipient vessels at the anastomotic sites. The saphenous vein graft was prepared by lysing the valves with a valvulotome. The blood flow could then be reversed in the vein, allowing it to be used in either direction as a bypass graft. An illustrative case including angiograms that confirm good patency and blood flow through the reversed-flow bypass graft is presented. It is concluded that the reversed-flow saphenous vein graft provides a more physiologically suitable conduit than standard vein grafts. Lysis of the valves allows the graft to be used in an orientation that takes advantage of the natural tapering of the vein to produce a better match with the recipient vessels at the anastomotic sites. Minimizing diameter changes at the proximal and distal anastomoses helps reduce turbulence, which has been implicated as a cause of early graft failure and thrombosis. PMID- 9347994 TI - Omental transplantation and spinal cord injury. PMID- 9347995 TI - Hemorrhage and shunting. PMID- 9347996 TI - Functional hemispherectomy. PMID- 9347997 TI - Internal carotid or ascending pharyngeal artery. PMID- 9347998 TI - Hypermetabolism or hyperglycolysis. PMID- 9348038 TI - The role of DNA demethylation during development. AB - The somatic genomic methylation pattern which plays a role in the suppression of basal gene activity is established anew in each generation through developmentally regulated de novo and demethylation steps. Demethylation appears to be carried out by a nucleotide exchange reaction which may involve RNA molecules, and is directed to specific loci in the genome through interactions between cis acting elements and trans acting factors. PMID- 9348040 TI - A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization. AB - BACKGROUND: In many respects Archaea are much more like eukaryotes than prokaryotes with respect to the conservation of many of the components involved in transcription, translation and DNA replication. So far, only a few DNA polymerases with structures similar to those of eukaryotic DNA polymerase a have been found in Archaea. The identification and characterization of all the DNA polymerases of one archaeon would add considerably to our knowledge of the basic mechanisms of DNA replication in these organisms. RESULTS: We have identified a novel DNA polymerase composed of two proteins, DP1 and DP2, with molecular weights of 69294 Da and 143161 Da, respectively, in the hyperthermophilic archaeon, Pyrococcus furiosus, and have cloned the corresponding genes which are tandemly arranged on the Pyrococcus genome. No significant sequence homology was found between these two proteins and other known DNA polymerases. The pol genes were transcribed as part of a single operon that additionally contained genes homologous to the cdc18+/CDC6 and Dmc1/Rad51 family of proteins. We purified the Pyrococcus DNA polymerase from Escherichia coli strains expressing the cloned genes and characterized its activity. It possesses strong 3' --> 5' exonucleolytic activity and has a template-primer preference which is characteristic of a replicative DNA polymerase. CONCLUSION: In P. furiosus, we identified a second DNA polymerase encoded by two genes, neither of which display significant homology to any other known DNA polymerase. Both the enzymatic properties of the enzyme and the gene organization raise the possibility that this enzyme might be the replicative DNA polymerase of P. furiosus. PMID- 9348039 TI - Switching yeast from meiosis to mitosis: double-strand break repair, recombination and synaptonemal complex. AB - BACKGROUND: When Saccharomyces cerevisiae cells that have begun meiosis are transferred to mitotic growth conditions ('return-to-growth', RTG), they can complete recombination at high meiotic frequencies, but undergo mitotic cell division and remain diploid. It was not known how meiotic recombination intermediates are repaired following RTG. Using molecular and cytological methods, we investigated whether the usual meiotic apparatus could repair meiotically induced DSBs during RTG, or whether other mechanisms are invoked when the developmental context changes. RESULTS: Upon RTG, the rapid disappearance of meiotic features--double-strand breaks in DNA (DSBs), synaptonemal complex (SC), and SC related structures-was striking. In wild-type diploids, the repair of meiotic DSBs during RTG was quick and efficient, resulting in homologous recombination. Kinetic analysis of double-strand breakage and recombination indicated that meiotic DSB formation precedes the commitment to meiotic levels of recombination. DSBs were repaired in RTG in dmc1, but not rad51 mutants, hence repair did not occur by the usual meiotic mechanism which requires the Dmc1 gene product. In haploids, DSBs were also repaired quickly and efficiently upon RTG, showing that DSB repair did not require the presence of a homologous chromosome. In all strains examined, SC and related structures were not required for DSB repair or recombination following RTG. CONCLUSIONS: At least two pathways of DSB repair, which differ from the primary meiotic pathway(s), can occur during RTG: One involving interhomologue recombination, and another involving sister chromatid exchange. DSB formation precedes commitment to recombination. SC elements appear to prevent sister chromatid exchange in meiosis. PMID- 9348041 TI - Two closely-related left-right asymmetrically expressed genes, lefty-1 and lefty 2: their distinct expression domains, chromosomal linkage and direct neuralizing activity in Xenopus embryos. AB - BACKGROUND: Vertebrates have numerous lateral asymmetries in the position of their organs, but the molecular basis for the determination of left-right (L-R) asymmetries remains largely unknown. TGFbeta-related genes such as lefty and nodal are L-R asymmetrically expressed in developing mouse embryos, and may be involved in L-R determination. RESULTS: We have identified two highly conserved genes, lefty-1 and lefty-2, in the mouse genome. These two genes are tightly linked on mouse chromosome 1. lefty-1 and lefty-2 are both expressed in a L-R asymmetric fashion in mouse embryos. However, the major expression domains of the two genes are different: lefty-1 expression is predominantly confied to the left side of ventral neural tube, whereas lefty-2 is strongly expressed in the lateral plate mesoderm on the left side. In embryos homozygous for the iv and inv mutation, which cause situs inversus, the expression sites of both genes are affected, either reversed or bilaterally, indicating that lefty-1 and lefty-2 are downstream of iv and inv. Although Lefty-1 and Lefty-2 prepro-proteins are not readily processed in cultured cells, BMP2-Lefty chimeric proteins can be processed to a secreted form. We have examined the activities of Lefty-1 and Lefty-2 in Xenopus embryos. In animal cap explants, Lefty-1 and Lefty-2 induce neural cells in the absence of mesoderm induction. The direct neuralizing activities of Lefty-1 and Lefty-2 thus seem remarkably similar to those of BMP antagonists such as noggin and chordin, suggesting that the action of Lefty-1 and Lefty-2 may be to locally antagonize BMP (bone morphogenic protein)-mediated signals in tissues positioned on the left side of the mouse embryos. CONCLUSION: There are two lefty genes in mice (lefty-1 and lefty-2), both of which are expressed in a L-R asymmetric fashion and are downstream of iv and inv. Lefty-1 and Lefty-2 possess direct neuralizing activity in Xenopus embryos, resembling the activities of BMP antagonists. PMID- 9348042 TI - Chi-star, a chi-related 11-mer sequence partially active in an E. coli recC1004 strain. AB - BACKGROUND: chi sequence (5'GCTGGTGG) of Escherichia coli was first identified as a site that increased the plaque size of bacteriophage lambda. Subsequent studies showed that this site is responsible for both the attenuation ofRecBCD exonuclease activity and the promotion of RecA, RecBCD-mediated recombination. It is known that bacteriophage lambda containing the chi site makes very small plaques on a recC* (recC1004) mutant because chi is not recognized by the RecBC*D mutant enzyme. RESULTS: We cloned E. coli chromosomal fragments in lambda which allowed lambda to form larger plaques on this recC1004 mutant. The fragments were found to share a chi-like 11-mer sequence, 5'GCTGGTGCTCG. Substitution of these fragments with a synthetic 11-mer of this sequence and single-base-pair substitution analysis of its last four nucleotides demonstrated that this sequence is both necessary and sufficient for the observed activity. The sequence, designated X* (chi-star), protected rolling-circle DNA replication in the recC1004 mutant and in the recBCD+ strain, most likely because it attenuated the exonuclease activity of the RecBC*D and RecBCD+ enzyme. chi-star, did not significantly stimulate lambda recombination in two assays. CONCLUSION: We have discovered that a mutant RecBCD enzyme responds, in vivo, to a longer chi variant. PMID- 9348044 TI - Pediatric death: managing the aftermath in the emergency department. AB - Managing the family conference in the emergency department after the sudden death of a child is difficult and, when mishandled, can be deleterious to the patient's family. We surveyed parents of children who died in an emergency department setting in an effort to elicit information that will help emergency physicians tell parents that their child has died. A 24-question survey was distributed to 60 parents identified by the Illinois chapter of the Sudden Infant Death Syndrome Alliance. Results were included if the parents indicated that their child was pronounced dead on arrival or died in an emergency department. If the victim was over 16 years of age, had a terminal illness, or died after hospitalization, results were excluded. Thirty-seven parents completed the survey. Seventy-six percent felt that it was the attending physician's responsibility to inform the family that the child had died. Holding the dead child was helpful for 88%. Seventy-six percent felt that being asked about organ donation would not be offensive. For most parents, having the child's clothes returned was important. Fully 92% of the respondents would have liked a physical memento of the child, such as a print or mold of the child's hand or a lock of hair. Most parents felt that a follow-up telephone call would be helpful, although only a small minority received such a call. Parents whose child died in an emergency department provided some concrete suggestions for emergency physicians regarding informing parents that their child died. Although the majority of children died of sudden infant death syndrome, the results may be applicable to other pediatric deaths. PMID- 9348043 TI - Establishing an approach to syncope in the emergency department. AB - A pilot study was conducted to enumerate the most common evaluations done in the emergency department (ED) of a community hospital in assessing patients presenting with a first episode of syncope and to determine the feasibility of defining a clinically useful set of investigations to identify the subset of syncopal patients that can be safely discharged from the ED. The study was conducted as a retrospective chart review of patients seen during an 8 week period. In the course of the study, 33 consecutive adult patients presenting to the ED with first episodes of syncope were identified. Patients were excluded if they had previous recurrent syncopal episodes or a known disorder leading to syncope. ED charts of the participants were reviewed to determine the types of investigations for syncope done by unblinded emergency physicians, which investigations showed abnormal results, and which were useful in determining etiology of syncope or in deciding which patients needed admission. The average number of investigations performed on each patient was 7 +/- 4, with a range of 1 17 investigations. Twelve percent of syncopal patients (4/33) were deemed, retrospectively, to have required hospital admission based on a review of their charts and follow-up interviews. Without specific clinical indicators, laboratory and radiologic investigations were not useful in determining either the etiology of the syncopal episode or the need for admission. In this small study, few patients presenting with new onset syncope required admission. The number and types of investigations performed on these patients was inconsistent. Further study is needed to determine whether syncopal patients requiring admission can be identified in the ED with a small number of standard inexpensive laboratory investigations. PMID- 9348045 TI - Denial of emergency department authorization of potentially high-risk patients by managed care. AB - This study was designed to evaluate patients presenting to a large urban university emergency department (ED) who were subsequently denied authorization for reimbursed care by their managed care provider and to characterize the denial as potentially safe or unsafe based on published triage criteria. A consecutive case surveillance was performed from October 1, 1994 to September 30, 1995 at a university-based ED (30,000 visits per year) for adult patients in inner-city Chicago. Cases were comprised of adult managed care participants whose providers refused by telephone to authorize payment for ED services and who then left the ED without treatment. Chief complaints and vital signs were used to categorize patients as high-risk or nonemergent based on previously published criteria. A total of 2,965 adult managed care patients presented to the ED during the study period, representing 11.1% of the total ED census. Of these patients, 244 (8.2%) were denied authorization for payment of their care. By previously established criteria, 115 (47.1%) were identified as potentially unstable, 61 (53%) due to abnormal vital signs and 54 (47%) with other high-risk indications such as severe pain, chest pain, or abdominal pain. These potentially high-risk patients may subsequently suffer adverse outcomes. Current guidelines used for telephone triage by managed care to divert patients from our ED do not meet previously published safe triage criteria. PMID- 9348046 TI - Patients' satisfaction when denied authorization for emergency department care by their managed care plan. AB - We conducted a survey of managed care plan (MCP) patients who presented to the emergency department (ED) but were denied insurance authorization during a 3 month period. Patients were identified by triage or registration records, contacted by telephone after their visit, and surveyed regarding their satisfaction with the ED and MCP, follow-up care, and future behavior. We surveyed 72 (73.4%) of 98 subjects who were denied authorization. Forty-nine (68.1%) were redirected to a clinic or primary physician, 14 (19.4%) to an urgent care or other ED, and 9 (12.5%) were given no follow-up. Fifty-five respondents (76.4%) stated they had followed-up as directed, but 34 (47.2%) felt the delay had a negative impact. Thirty-nine (54.2%) were dissatisfied with their MCP. If their problems were to recur, 27 (37.5%) stated they would go to a clinic or call their MCP, but 34 (47.2%) would return to the ED. Many patients who are denied authorization are dissatisfied with their MCP and will return to the ED in the future, despite previous denials. PMID- 9348047 TI - Headache in the emergency department: importance of history in identifying secondary etiologies. AB - Headache is a common emergency department complaint and has a broad differential diagnosis. Most commonly the headache is without serious underlying cause, but occasionally can be the manifestation of a more catastrophic illness. History may be the most important aspect in the evaluation of headache patients and careful attention to historical clues or atypical symptoms may point to a diagnosis. We present a patient with a first-time headache, which was ultimately found to be secondary to a carotid cavernous fistula. Historical features that require a more aggressive work-up of headache are discussed. PMID- 9348048 TI - Psoas abscess with sepsis mimicking traumatic hemorrhagic shock after a fall. AB - Abscess of the psoas muscle is infrequently encountered. An infectious emergency of this type usually presents in a nonspecific manner and thus poses a significant diagnostic challenge to the emergency physician. Diagnosis and specific treatment are often delayed, which can lead to increased mortality. This case report presents a patient with altered mental status and hypotension after a fall, who was initially managed as a trauma victim. Emergency department evaluation initially focused on a traumatic etiology of the above abnormalities. Subsequent assessment determined that the patient's condition was due to an underlying psoas abscess with sepsis. Appropriate anatomy, clinical presentation, and management are discussed in hopes of increasing physician awareness of this uncommon infectious condition. PMID- 9348049 TI - Successful treatment of severe hypothermia and prolonged cardiac arrest with closed thoracic cavity lavage. AB - This case report describes the resuscitation of a 19-year-old man who had been immersed in ice water for 14 h and presented with a rectal temperature of 22 degrees C and no pulses. It reports the successful use of prolonged cardiac massage (3.5 h) and closed thoracic cavity lavage in the treatment of severe hypothermia. It also confirms that victims of severe hypothermia can be effectively treated in peripheral hospitals not equipped for cardiopulmonary bypass. PMID- 9348051 TI - Lingual ischemia following tongue entrapment in a glass bottle. AB - A 10-year-old, previously healthy female presented to the emergency department via emergency medical service transport, with her tongue tightly entrapped inside a glass bottle (9 oz, Yoohoo brand of chocolate drink). The tongue was massively edematous and ecchymotic due to impaired venous return from constriction by the neck of the bottle. After repeated attempts at mechanically reducing the tongue out of the bottle, a professional glazier was contacted, who was able to remove the bottle in the operating room with a steel glass cutter. Needle evacuation of a small hematoma was then performed to decrease the pressure ischemia to the tongue, which began to improve quickly. PMID- 9348050 TI - An occult cause of persistent nausea and vomiting. AB - We report a patient with multiple negative evaluations during emergency department visits and inpatient admissions for unexplained, intermittent nausea, vomiting, and abdominal pain. The etiology of her symptoms was not revealed until her 13th hospital visit, when head magnetic resonance imaging suggested active neurocysticercosis. Central etiologies should be considered for intractable nausea and vomiting in neurologically intact patients even if head computed assisted tomography scan is negative. PMID- 9348052 TI - Spontaneous pneumomediastinum secondary to hyperemesis gravidarum. AB - A case of spontaneous pneumomediastinum secondary to hyperemesis gravidarum is presented. The pathophysiology, clinical presentation, differential diagnosis, and management of this unusual complication of hyperemesis gravidarum are reviewed. PMID- 9348053 TI - Acute pancreatitis secondary to ifosfamide. AB - Acute pancreatitis in cancer patients can be secondary to the malignant process itself. It is also a rare complication of antineoplastic agent administration. Ifosfamide is an effective drug in the treatment of several tumors and has known neurologic, renal, and hematologic toxicities. There is only one recent report in the literature of pancreatitis associated with ifosfamide. We report a case of a 65-year-old woman with small cell bronchogenic carcinoma without pancreatic metastases who developed acute pancreatitis after ifosfamide administration. PMID- 9348054 TI - A case of paraquat poisoning and subsequent fatality presenting to an emergency department. AB - Paraquat (1,1'-dimethyl-4,4'-dipyridylium) is an herbicide associated with both accidental and intentional ingestion, leading to severe and often fatal toxicity. Prognosis is largely dependent on the amount of paraquat absorbed. Rapid identification of the symptoms of paraquat toxicity (burns or ulceration at the site of ingestion or injection, acute respiratory distress, and renal failure) can facilitate early treatment intervention to limit absorption. We report a case of a 71-year-old man with a suicidal ingestion of paraquat 2 days prior to presentation. Serum paraquat levels, time elapsed since ingestion, and clinical symptoms all indicated poor prognosis. The patient developed severe respiratory distress and progressive renal failure, and died 6 days after admission to the hospital. PMID- 9348055 TI - Ethylene glycol poisoning: case report of a record-high level and a review. AB - Ethylene glycol is commonly found in automobile antifreeze and a variety of other commercial products. Ingestion of ethylene glycol, either accidentally or in a suicide attempt, is characterized by severe acidosis, calcium oxalate crystal formation and deposition, and a wide variety of end organ effects that may be fatal. We present a case of a patient who ingested a massive amount of ethylene glycol in a suicide attempt and yet survived with minimal sequelae. A comprehensive review of the literature on the pathology and pathophysiology of ethylene glycol toxicity on each organ system is provided, along with information on diagnosis and current treatment recommendations. PMID- 9348056 TI - Metacarpophalangeal joint dislocation: indications for open surgical reduction. AB - In complex dislocations of the metacarpophalangeal joint, the volar plate is separated from the proximal phalanx and the metacarpal head is entrapped within surrounding tissue structures. These complex dislocations must be managed by open surgical reduction to reduce the dislocation and realign the volar plate. A 58 year-old male presented to the emergency department with a complex dislocation of the metacarpophalangeal joint of the left little finger, which was successfully treated by open reduction in the operating room. The indications for open reduction of metacarpophalangeal joint dislocations are reviewed. PMID- 9348057 TI - Emergency department repair of hand lacerations using absorbable vicryl sutures. AB - The use of absorbable suture material has a number of potential advantages when compared to nonabsorbable suture. We conducted a 5-year retrospective study of 102 patients with hand lacerations and compared the quality of scar formation and healing in these patients. Those patients who did not have tendon, nerve, or bone injury were included in the study. Lacerations were repaired with either 5-0 Vicryl or nylon. There were no reported complications or infections in any study group patient. The quality of scar, when compared visually and by palpation, was the same at the end of 6 months. In addition, there was no difference in the incidence of scar retraction. We conclude that the use of absorbable suture material is an acceptable alternative in the repair of hand lacerations. PMID- 9348058 TI - Molten metal burns: further evidence of industrial foundries' failure to comply with Occupational Safety and Health Administration regulations. AB - The purpose of this report is to describe another case of a molten metal burn to the foot of a foundry worker. The foundry in which he worked failed to comply with Occupational Safety and Health Administration regulations with regard to protective apparel. This injury could have been prevented with annual, unscheduled inspections by the Occupational Safety and Health Administration and with enforcement of additional regulations regarding protective apparel. PMID- 9348059 TI - Biomechanical performance of cutting edge surgical needles. AB - The purpose of this study was to compare the biomechanical performance of cutting edge needles made of S45500 stainless steel alloy to Surgalloy stainless steel. The new high-nickel stainless steel alloy, Surgalloy, has superior performance characteristics over that of the other high-nickel stainless steel alloy, S45500. The Surgalloy needle is produced from a stronger stainless steel alloy than the S45500 needle. The Surgalloy needle has considerably greater resistance to bending than the needle produced from S45500 alloy. In addition, Surgalloy stainless steel has almost a twofold greater resistance to fracture than the S45500 stainless steel alloy. PMID- 9348060 TI - Laboratory testing in ethanol, methanol, ethylene glycol, and isopropanol toxicities. AB - Toxicity from ethanol, methanol, ethylene glycol, and isopropyl alcohol varies widely, and appropriate use of the available laboratory tests can aid in timely and specific treatment. Available testing includes direct measurements of serum levels of these alcohols; however, these levels often are not available rapidly enough for clinical decision making. This article discusses the indications and methods for both direct and indirect testing for ethanol, methanol, ethylene glycol, and isopropanol toxicity. Also discussed are the costs, availability, and turn-around times for these tests. PMID- 9348061 TI - An elderly patient with myalgias. PMID- 9348062 TI - The chest radiograph we should never see. PMID- 9348063 TI - From Creutzfeldt-Jakob disease to the mad cow epidemic. AB - Hans-Gerhard Creutzfeldt and Alfons Jakob independently authored clinical and pathologic descriptions of a new syndrome in the 1920s. This syndrome, which subsequently came to be named after them, was characterized by dementia, motor and coordination abnormalities, a fatal course, and pathologic findings of diffuse spongiform neuronal degeneration. Although it appeared for many years to be little more than a medical curiosity, Creutzfeldt-Jakob disease attained widespread attention by its pathologic similarity to kuru and bovine spongiform encephalopathy, "mad cow disease." Because there are sporadic, familial, and iatrogenic forms of Creutzfeldt-Jakob disease, it is considered to have both genetic and infectious aspects. Although its causation has for some time been ascribed to "slow viruses," the etiology of Creutzfeldt-Jakob disease is currently thought to be due to prions, small proteinaceous infectious particles that have genetic encoding. The debate regarding whether the appearance of atypical Creutzfeldt-Jakob disease can be linked to the epidemic of "mad cow disease" is currently unresolved. PMID- 9348064 TI - David R. Boyd Lecture in Trauma Care and Emergency Medical Systems: introductory remarks. PMID- 9348065 TI - David R. Boyd Lecture in Trauma Care and Emergency Medical Systems: upper extremity injuries--past, present, and future. AB - Since its recognition as a subspeciality soon after World War I, hand surgery has evolved into a highly specialized and sophisticated field of medicine. Enormous advances have been made in the diagnosis and treatment of disorders of the upper limb; primary tenorrhaphy and the application of microsurgical techniques for revascularization, replantation, and free tissue transfer are among the most important innovations. This article reviews the history of flexor tendon surgery and reconstructive microsurgery up to the present time, and makes some predictions for the future direction of these, and other, areas of hand surgery. PMID- 9348066 TI - The prion paradox. PMID- 9348067 TI - The application of continuous quality improvement (CQI) principles to emergency medicine procedures. PMID- 9348068 TI - The utility of creatine testing prior to i.v. contrast studies in the emergency department. PMID- 9348069 TI - The critical patient who refuses treatment: an ethical dilemma. AB - In clinical practice, emergency physicians must often make decisions in their patients' best interests when the patients are unable to do so themselves. The usual requirement for informed consent stems from recognizing individuals' autonomy and their right to make decisions affecting their bodies. Abandoning a requirement for consent is an emergency exception to the ethical and legal principles and comes into play only when a person lacks decision-making capacity. In some instances, it may be unclear whether a patient has this capacity, confounding the physician's management decisions. How should emergency physicians assess a patient's decision-making capacity? When may they ethically and legally override a patient's expressed desire for treatment or nontreatment? These issues are discussed in the context of an actual case. PMID- 9348071 TI - Recommendations for the management of rural, remote, and isolated emergency health care facilities in Canada. Canadian Association of Emergency Physicians. PMID- 9348070 TI - Root cause analysis of laboratory delays to an emergency department. AB - The Q-Probes study has identified benchmark interinstitutional laboratory median turnaround times (TAT) of 25 min for hemoglobin and 36 min for potassium. Our objectives were to measure the emergency department (ED)/laboratory TAT and other relevant laboratory processing and reporting times, and to identify root causes of laboratory delay. A flow chart was developed for the ordering, collecting, analyzing, and reporting of laboratory results. Time intervals were prospectively recorded for complete blood count (CBC) and K+ in a cross-sectional study, using the flow chart, and defined as follows: TAT was the interval from blood draw (BD) to ED report; BD time was the interval from order processing to BD; and order processing time was the interval from physician ordering to the unit coordinator processing the orders. Median times with interquartile ranges are reported. CBC TAT was 38 min (29-51.5), and K+ TAT 58 min (45-76.5). Order processing time was 7 min (4-15). The laboratory assistant BD time was 17 min (8-30) for CBC and 15 min (7.75-32.25) for K+ as compared to 0 min for a nurse, yet the venipuncture method (laboratory assistant technique) had a recollection rate of 1% (1/93) due to hemolysis vs. 20% (19/95) via the i.v. catheter (nurse technique). Of stat ED blood work, 24% was for admitted patients held in the ED. Laboratory reporting times are delayed with these root causes: laboratory assistant availability; recollection rate; volume of tests for ED admitted patients; and order processing time. PMID- 9348072 TI - Multiple functions of general transcription factors TFIIE and TFIIH in transcription: possible points of regulation by trans-acting factors. AB - General transcription factors together with RNA polymerase II assemble on the promoter DNA and initiate transcription accurately in response to a variety of signals. Such signals enhance preinitiation complex formation by targeting components thereof via several alternative pathways. Two components of the initiation complexes, TFIIE and TFIIH, are known to function at both a late stage of transcription initiation and the following promoter clearance. TFIIH has been studied extensively because of its multiple enzymatic activities, functioning not only in transcription but also in nucleotide excision repair and cell cycle control. Fewer data have been reported for TFIIE, but its potential regulatory function as to TFIIH warrants further attention. In this review, an overall perspective of the functional roles of TFIIE and TFIIH during transcription initiation and the following promoter clearance will be presented as it has emerged from recent studies. PMID- 9348073 TI - Effects of fibronectin fragments on DNA transfection into mammalian cells by electroporation. AB - Two kinds of cells, human epidermoid carcinoma A431 cells and simian kidney COS-7 cells, were transfected with the chloramphenicol acetyl transferase (CAT) gene by electroporation, and then cultivated on culture dishes coated with two different forms of recombinant fibronectin fragments consisting of cell-binding domain (C 274) or heparin-binding domain and CS1 region (H-296). In the case of A431 cells, H-296-coated dishes significantly increased the amount of expressed CAT and the adhesion of electroporated cells in comparison with non-coated dishes. C-274 was effective for COS-7 cells. Overall, these fibronectin fragments increased the recovery of the transfectants on A-431 cells and COS-7 cells, respectively. PMID- 9348074 TI - Perinuclear membrane localization of alphaKAP, a protein produced from a gene within the gene of calmodulin-dependent protein kinase IIalpha. AB - AlphaKAP is a protein produced from a gene within the gene of the a isoform of calmodulin-dependent protein kinase II (CaM-kinase IIa). It consists of the association domain of CaM-kinase IIa and a highly hydrophobic amino-terminal stretch consisting of 25 amino acids which is absent from CaM-kinase IIalpha. We previously demonstrated that alphaKAP is an integral membrane protein by subcellular fractionation analysis [Sugai, R., Takeuchi, M., Okuno, S., and Fujisawa, H. (1996) J. Biochem. 120, 773-779], but the exact subcellular localization of alphaKAP was not well understood. Here we demonstrate that alphaKAP is localized on the nuclear membrane of COS-7 cells transiently expressing alphaKAP. The nuclear membrane and perinuclear small vesicles were immunostained with an antibody against a synthetic peptide corresponding to the carboxyl-terminal 15 amino acids of alphaKAP. In contrast to the intact alphaKAP, the mutant alphaKAP, from which the hydrophobic amino-terminal segment had been deleted, accumulated within nuclei. Thus, alphaKAP may function as an anchoring protein for CaM-kinase II and/or other proteins in the perinuclear membrane. PMID- 9348075 TI - 2APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable modulator of Ins(1,4,5)P3-induced Ca2+ release. AB - The effects of a novel membrane-penetrable modulator, 2APB (2-aminoethoxy diphenyl borate), on Ins(1,4,5)P3-induced Ca2+ release were examined. 2APB inhibited Ins(1,4,5)P3-induced Ca2+ release from rat cerebellar microsomal preparations without affecting [3H]Ins(1,4,5)P3 binding to its receptor. The IC50 value (concentration producing 50% inhibition) of 2APB for inhibition of Ins(1,4,5)P3 (100 nM) induced Ca2+ release was 42 microM. Further increase in the concentration of 2APB (more than 90 microM) caused a gradual release of Ca2+ from cerebellar microsomal preparations. Addition of 2APB to the extracellular environment inhibited the cytosolic Ca2+ ([Ca2+]c) rise in intact cells such as human platelets and neutrophils stimulated by thromboxane-mimetic STA2 or thrombin, and leukotriene B4 (LTB4) or formyl-methionine-leucine-phenylalanine (FMLP), respectively. 2APB inhibited the contraction of thoracic aorta isolated from rabbits induced by angiotensin II (AII), STA2, and norepinephrine in a non competitive manner, but showed no effect on the contraction of potassium depolarized muscle. 2APB had no effect on the Ca2+ release from the ryanodine sensitive Ca2+ store prepared from rat leg skeletal muscle and heart. Although the specificity of 2APB with respect to the intracellular signaling system was not fully established, 2APB is the first candidate for a membrane-penetrable modulator of Ins(1,4,5)P3 receptor, and it should be a useful tool to investigate the physiological role of the Ins(1,4,5)P3 receptor in various cells. PMID- 9348076 TI - Implication of protein kinase C-alpha, delta, and epsilon isoforms in ischemic preconditioning in perfused rat hearts. AB - Ischemic preconditioning is a phenomenon in which one or several cycle(s) of brief ischemia-reperfusion protects the myocardium against the cell injury caused by subsequent prolonged ischemia. Protein kinase C (PKC) inhibitors blunt the cardioprotection arising from ischemic preconditioning. To investigate which PKC isoform is involved in ischemic preconditioning, we identified the PKC isoform that translocates to the membrane fraction by means of immunoblotting with specific antibodies. PKC-alpha, delta, epsilon isoforms all increased in the membrane fraction after three cycles of 3 min ischemia and 5 min reperfusion (ischemic preconditioning) in the perfused rat heart. The ischemic preconditioning significantly improved the recovery of left ventricular developed pressure (LVDP) during reperfusion following 20 min of ischemia. A PKC specific inhibitor, chelerythrine (1.0 microM) blocked the effect of ischemic preconditioning on LVDP recovery and the translocation of PKC-alpha, delta, epsilon isoforms. These data suggest that one or more of these three isoforms of PKC is involved in ischemic preconditioning by phosphorylating membrane proteins. PMID- 9348077 TI - Analysis of the stability of mutant lysozymes at position 15 using X-ray crystallography. AB - His 15 of hen lysozyme is located at the protein surface and is partly buried by the neighboring residues. The side chain of His 15 forms hydrogen bonds with surrounding residues and these hydrogen bonds are somewhat buried. A series of mutant lysozymes at the position 15 (Gly, Ala, Val, and Phe) was prepared, and their stabilities were analyzed by GdnHCl denaturation and X-ray crystallography. The mutants were less stable than the wild type at pH 5.5 and 35 degrees C. In H15G and H15A, X-ray crystallography revealed two fixed water molecules at the mutated region, which formed similar hydrogen bonds to those in the wild type. On the other hand, it was suggested that the hydrogen bonds were disrupted and that several unfavorable van der Waals' contacts occurred in H15V and H15F. Therefore, we concluded that His 15 stabilized the lysozyme structure by forming hydrogen bonds and the best packing with the neighboring residues. Moreover, we found that the method of protein stabilization by increasing the hydrophobicity of an amino acid residue was not always effectively applicable, especially when the residue had formed a hydrogen bond. PMID- 9348078 TI - Isolation of cleavage furrows from eggs of regular sea urchins and identification of furrow-specific proteins. AB - We have developed a method for the isolation of cleavage furrows from dividing sea urchin eggs, which is applicable to various sea urchin species. The new method differs from that used for isolating cleavage furrows from sand dollar Clypeaster japonicus eggs [Yonemura, S., Mabuchi, I., and Tsukita, S. (1991) J. Cell Sci. 100, 73-84] in the type and concentration of detergent included in the isolation medium, the temperature during the treatment of dividing eggs with the isolation medium, and the centrifugation conditions. The contractile ring was included in the isolated cleavage furrows, as seen on rhodamine-phalloidin staining of actin filaments. When the furrows were isolated with the isolation medium containing both NaF and beta-glycerophosphate, which are potent protein phosphatase inhibitors, the isolated furrows were found to be accompanied by the mitotic apparatus. When the isolation was carried out in the absence of both NaF and beta-glycerophosphate, cleavage furrows without the mitotic apparatus were obtained. The development of a method of isolation of cleavage furrows from regular sea urchin eggs enabled us to compare protein constituents among furrows from different sea urchin and sand dollar species. We found that 32, 36, and 51 kDa proteins were concentrated in common in the cleavage furrows isolated from eggs of the sand dollars, C. japonicus and Scaphechinus mirabilis, and the sea urchins, Hemicentrotus pulcherrimus and Strongylocentrotus nudus, on two dimensional gel electrophoreses. PMID- 9348079 TI - Identification and subcellular localization of a novel mammalian dynamin-related protein homologous to yeast Vps1p and Dnm1p. AB - The dynamin family of GTP-binding proteins are implicated in vesicular transport. These include mammalian dynamins I, II, III, and yeast Vps1p and Dnm1p. Dynamin is involved in the formation of clathrin-coated vesicles at the plasma membrane. On the other hand, Vps1p and Dnm1p appear to be involved in transport from the late Golgi compartment to vacuoles and in an endocytic process, respectively. In this study, we identified a novel human protein, named Dnm1p/Vps1p-like protein (DVLP). It resembled more closely Dnm1p and Vps1p than dynamins not only in the primary structure but also in the domain organization. DVLP mRNA was expressed ubiquitously, suggesting that this protein plays a fundamental role in cellular function. Immunofluorescence analysis of cells expressing epitope-tagged DVLP revealed that it showed a diffused perinuclear staining pattern that was not superimposed on that of the marker protein for the Golgi apparatus, trans-Golgi network, lysosomes, endosomes, or endoplasmic reticulum. These data suggest that DVLP is not involved in the formation of known coated vesicles. PMID- 9348080 TI - Purification of bovine soluble guanylate cyclase and ADP-ribosylation on its small subunit by bacterial toxins. AB - Soluble guanylate cyclase (sGC) consisting of two different subunits (alpha: Mr = 74,000, beta: Mr = 69,000) was purified more than 12,000-fold in terms of specific activity from the supernatant of bovine lung homogenates and characterized. The heme content determined with the pyridine hemochromogen method and Bradford's protein assay was 0.8 heme per dimer. Cholera, pertussis, and botulinum C3 toxins modified exclusively the beta-subunit of sGC, yielding the ADP-ribose-bound compound with 1:1 stoichiometry, and Vmax for the cyclase reaction was increased 10 times by this modification. When the ADP-ribosylation of sGC was performed simultaneously with two or three bacterial toxins which have distinct amino acid specificities, the resultant enzyme had only one ADP-ribose, and the activity was the same as that of the enzyme modified with one toxin. When NO was incorporated into the reaction mixture containing the ADP-ribosylated sGC, the cyclase activity noticeably increased by approximately the same amount as that seen for the unmodified enzyme. Such effects were not seen with CO. When ADP ribosylated sGC was incubated with Mn2+, the enzyme activity was synergistically increased. The heme-deleted sGC was also ADP-ribosylated by bacterial toxins and its activity was raised. These findings suggest that sGC has an ADP-ribosylation site near the GTP binding site, like other GTP-binding proteins, and that the beta-subunit regulates the activity. PMID- 9348081 TI - Purification and characterization of polyamine aminotransferase of Arthrobacter sp. TMP-1. AB - Polyamine aminotransferase of Arthrobacter sp. TMP-1 was induced by 1,3 diaminopropane (DAP), N-3-aminopropyl-1,3-diaminopropane (norspermidine), spermidine, and spermine, but not by putrescine. The enzyme was purified to homogeneity. Its molecular weight and subunit size were 129,000 and 64,000, respectively. Its absorption spectrum had maxima at 280 and 420 nm and a shoulder at about 350 nm, and changes were observed upon the addition of DAP, putrescine, and sodium borohydride. The spectrum and its changes indicated that the enzyme contained pyridoxal-5'-phosphate as the coenzyme. The coenzyme content was found to be 1 mol per mol of subunit. DAP, putrescine, norspermidine, spermidine, and spermine were active amino donors and gave relative rates of 100, 73, 24, 30, and 23%, respectively. Pyruvate was the most active amino acceptor, while 2 ketoglutarate and oxaloacetate were inert. The equilibrium constant of the DAP pyruvate transamination was 0.34. DAP was suggested to be a minor product of the norspermidine-pyruvate reaction. PMID- 9348082 TI - Action of polyamine aminotransferase on norspermidine. AB - The norspermidine-pyruvate reaction catalyzed by polyamine aminotransferase from Arthrobacter sp. TMP-1 formed N-3-aminopropyl-3-aminopropionaldehyde (APAPAL), L alanine, 1,3-diaminopropane (DAP), allylamine, and acrolein, and the relative rates of formation of the latter four products were 24, 3.3, 2.3, and 1.2%, respectively, of the rate of the DAP-pyruvate transamination. The identification of APAPAL was done by 13C-NMR after it had been enzymatically oxidized to N-3 aminopropyl-beta-alanine followed by isolation of the oxidized product. The DAP was also isolated and identified by 13C-NMR. The allylamine and acrolein were identified by HPLC and a specific color reaction with m-aminophenol, respectively. In the absence of pyruvate, the enzyme catalyzed the elimination of DAP from norspermidine to yield allylamine, and the addition of DAP to allylamine to yield norspermidine with relative rates of 0.007 and 0.095%, respectively. When allylamine was incubated with the enzyme as the sole substrate, it was converted to N-allyl-1,3-diaminopropane and an unidentified product. PMID- 9348084 TI - Hairpin structure of an RNA 28-mer, which contains a sequence of the enzyme component of a hammerhead ribozyme system: evidence for tandem G:A pairs that are not of side-by-side type. AB - An RNA 28-mer (Rz28) was obtained as a major product by in vitro transcription with T7 RNA polymerase of a promoter-template DNA, which contains a sequence for the enzyme component, RNA 24-mer (Rz24), of a mutant hammerhead ribozyme system. Sequence analysis and enzymatic probing study showed that Rz28 has 4 extra nucleotides at the 3'-terminus, the sequence of which is complementary to that of the 5'-terminal sequence of Rz24, and forms a stable hairpin structure. NMR studies using a 15N-guanine-labeled derivative suggested that Rz28 contains tandem G:A pairs that are not of the side-by-side type which is found in the crystal structure of hammerhead ribozyme complexes. Comparison of the HMQC spectra of 15N-guanine-labeled Rz28 and Rz24 suggested that Rz24 also contains the same type of tandem G:A pairs. PMID- 9348083 TI - Mechanisms of nitroso compound-induced inhibition of superoxide generation in neutrophils: fluorescence quenching of perylene by nitroso-compounds in the membrane fractions of neutrophils. AB - To investigate the mechanism of nitroso compound-induced inhibition of the respiratory burst in neutrophils, we studied fluorescence quenching of perylene by nitroso-compounds in the membrane fractions of neutrophils at 17, 27, and 37 degrees C and the reagent-induced inhibition of superoxide generation at 28 and 37 degrees C. With increasing temperature, the quenching of perylene fluorescence and inhibition of superoxide generation by nitrosobenzene (NB) were both diminished, while those by 2-nitrosotoluene (NT) were both enhanced. The temperature dependence of the inhibition constants and the quenching constants indicates that the binding of NB is exothermic (deltaH= -27 kJ/mol for inhibition and deltaH= -29 kJ/mol for quenching) and essentially enthalpy-driven. On the other hand, that of NT is endothermic (deltaH= +16 kJ/mol for inhibition and quenching) and essentially entropy-driven. Quenching studies of perylene fluorescence in synthetic vesicles made of endogenous polar lipids of neutrophils showed that the enthalpy changes of NB- and NT-binding with perylene in lipids were similar to each other. Moreover, their values were in good agreement with that of NT, but not of NB, in the membrane fractions, an assembly of proteins and lipids, of neutrophils. These results suggest that NB inhibits the activity by binding to proteins in the membrane, whereas inhibition by NT occurs through hydrophobic interaction with lipids and/or proteins. PMID- 9348085 TI - Myosin head interactions in Ca2+-activated skinned rabbit skeletal muscle fibers. AB - Interactions between the two myosin heads were studied in skinned rabbit slow twitch muscle fibers activated in the presence of vanadate (Vi), a phosphate analog. The strong complex between Vi, MgADP, and myosin trapped the myosin in an inactivated myosin x MgADP x Vi state. Electron paramagnetic resonance spectroscopy was used to quantitate the fraction of myosin heads trapped in the presence of a spin labeled analog of ATP (SLATP). Force was found to depend directly on the fraction of untrapped heads. At high [Vi] (low force), most untrapped heads would have a trapped partner. The equivalence of force with the proportion of untrapped heads shows that the isometric force produced by a single untrapped myosin head on a molecule with a trapped partner is equivalent to that produced by either head of a myosin molecule with neither head trapped. The actin activated MgATPase activities of one-headed and two-headed skeletal myosin species were inhibited similarly by Vi, suggesting that trapping one head did not preclude trapping its partner. These data indicate that the two skeletal muscle myosin heads can function without interacting during maximal Ca2+-activated force generation. PMID- 9348086 TI - Purification and properties of sn-glycerol-1-phosphate dehydrogenase from Methanobacterium thermoautotrophicum: characterization of the biosynthetic enzyme for the enantiomeric glycerophosphate backbone of ether polar lipids of Archaea. AB - The enzyme which seems to be responsible for the formation of the enantiomeric configuration of the glycerophosphate backbone (sn-glycerol-1-phosphate) of archaeal ether lipids was purified from a methanogenic archaeon, Methanobacterium thermoautotrophicum, and characterized. The enzyme, sn-glycerol-1-phosphate: NAD(P)+ oxidoreductase (sn-glycerol-1-phosphate dehydrogenase), was purified 7,600-fold from a cell free extract by ammonium sulfate fractionation and seven steps of chromatography. The final preparation exhibited a specific activity of 617 micromol/min/mg (Vmax) and gave a single band corresponding to 38 kDa on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The native enzyme showed an apparent molecular mass of 302 kDa on gel-filtration chromatography, indicating it is present as a homooctamer. Maximum activity was observed at 75 degrees C at near neutral pH. The activity was stimulated by potassium ions. The Km for dihydroxyacetone phosphate was 7.5 times smaller than that for sn-glycerol-1-phosphate, suggesting that the formation of sn-glycerol-1 phosphate is the natural direction in the cell. Under the assay conditions used, no product inhibition was observed. The N-terminal amino acid sequence was determined. PMID- 9348087 TI - Proteolytic cleavage sites of band 3 protein in alkali-treated membranes: fidelity of hydropathy prediction for band 3 protein. AB - To assess the fidelity of hydropathy prediction for band 3 protein, we determined the cleavage sites of the protein and the portions of the protein tightly bound to the membrane lipid bilayer by means of in situ proteolytic digestion. For the removal of all anticipated hydrophilic connector loops from membranes, we had to denature the band 3 protein molecule in situ by alkali treatment. When the alkali treated membranes were digested with trypsin, chymotrypsin, and pepsin, the majority of the anticipated transmembrane portions remained in the membrane fraction. However, five anticipated transmembrane portions were released into the supernatant fraction. Thus, the first, second, third, sixth and tenth anticipated transmembrane portions, in accordance with the hydropathy prediction, were released into the supernatant with the proteolytic digestion method. This indicates that these anticipated transmembrane portions are not bound with the boundary lipids although the hydrophobicity of these portions is comparable to that of the portions experimentally remaining in the membrane fraction. It is conceivable that the membrane peptide portions of band 3 protein could be classified into at least two categories, i.e. one bound to the boundary lipids and the other free from the boundary lipids. Approximately 90% of the transmembrane domain of the band 3 protein are recovered in either the supernatant fraction or the membrane fraction. The fidelity of hydropathy prediction for polytopic membrane proteins and the nature of the membrane embedded peptide portions are discussed. PMID- 9348088 TI - Difference in affinity for DNA between HMG proteins 1 and 2 determined by surface plasmon resonance measurements. AB - High mobility group (HMG) proteins 1 and 2 contain two similar but non-identical repeats of DNA-binding domains and an acidic C-terminal. The proposed functions of HMG proteins 1 and 2 imply a probable difference in their DNA-binding abilities. The primary studies by gel retardation assay showed that HMG2 has higher affinity than HMG1 for supercoiled and linear DNA. The DNA-binding of HMG2 appeared strong enough to allow exchange with HMG1 molecule already bound to DNA, while the DNA-binding region of HMG1 showed higher affinity than that of HMG2. In order to compare more quantitatively the affinities, surface plasmon resonance (SPR) measurements using a BIAcore instrument were conducted. The kinetic data indicated that the Kd for the complex of HMG2 with DNA is smaller than that of HMG1, in contrast to the situation for the DNA-binding region of these proteins. The sequence between the second DNA-binding domain and the acidic C-terminal of HMG proteins is required for tight DNA-binding. Also, the acidic C-terminal strongly modulates the DNA-binding ability of each protein. The usefulness of SPR measurement for quantitative analysis of affinity and regions involved in DNA binding under conditions nearly identical to those in solution is discussed. PMID- 9348090 TI - Prolyl aminopeptidase from Serratia marcescens: cloning of the enzyme gene and crystallization of the expressed enzyme. AB - We cloned and sequenced the Serratia marcescens prolyl aminopeptidase (SPAP) gene. Nucleotide sequence analysis revealed an open reading frame of 951 bp, encoding a protein of 317 amino acids with a predicted molecular weight of 36,083. The expressed enzyme was purified about 90-fold on columns of Toyopearl HW65C and DEAE-Toyopearl, with an activity recovery of 30%. The apparent molecular weight of the purified enzyme was 36,000 and 38,000 as estimated by SDS PAGE and gel filtration, respectively. The enzyme was not inhibited by diisopropyl phosphofluoridate (DFP) or phenylmethylsulfonyl fluoride (PMSF), but was markedly inhibited by 3,4-dichloroisocoumarin (DCIC). Crystals of the enzyme were grown by the hanging drop vapor diffusion method using PEG6000 as a precipitant at pH 6.5. The crystals are tetragonal with cell dimensions a= b =65.6 A, and c=169.8 A, a space group P4(1)2(1)2 or P4(3)2(1)2, and probably contain one monomer in the asymmetric unit. They diffract to at least 2.22 A resolution. PMID- 9348091 TI - Characterization of gramicidin S synthetase aggregation substance: control of gramicidin S synthesis by its product, gramicidin S. AB - An aggregation substance of gramicidin S synthetases was found and purified by DEAE-cellulose chromatography and CM-chromatography from cell debris of Bacillus brevis Nagano. It specifically aggregated and inactivated gramicidin S synthetases 1 (GS1) and 2 (GS2). On the basis of amino acid composition analysis, reversed-phase HPLC, FAB mass spectrometry, amino acid sequence analysis, and antibacterial activity, this substance (GrS-aggregation substance) was identified as gramicidin S. A gramicidin S derivative bearing a lysine residue in place of one ornithine residue was also detected as a minor component of GrS-aggregation substance. The extent of the aggregation was dependent on the concentration and relative amount of gramicidin S. The inhibition of the enzyme activities was irreversible and the inhibition was proportional to the amount of gramicidin S, like the aggregation of the enzymes. The degree of GS2 inhibition in the amino acid-dependent ATP-PPi exchange reaction varied with the amino acids of gramicidin S and increased in order of the amino acid sequence of gramicidin S. The degree of inhibition of the overall synthesis of gramicidin S was the same as that in the leucine-dependent exchange reaction. PMID- 9348089 TI - Involvement of interleukin-6 in activation of lysosomal cathepsin and atrophy of muscle fibers induced by intramuscular injection of turpentine oil in mice. AB - Serum IL-6 level increased after the injection of turpentine oil into the right gastrocnemius muscle in mice. The mRNA level of IL-6 was highest in the injected muscle at 12 h after injection, but was not identified in the opposite muscle. The activities of cathepsins B and B+L started to elevate after 12 h in the injected muscle and markedly increased after day 3. Likewise, the mRNA levels of cathepsins B and L markedly increased from day 1 to day 5 in the injected muscle. However, a very mild increase was also observed in the opposite muscle. Immunohistochemical staining of cathepsins B and L exhibited positive reactions as fine granules in myofibers at 12 h and strong positive reactions in the infiltrating macrophages at 3 days. Atrophy of myofibers type 1 and 2 was evident in a time-dependent manner in the injected muscle. Treatment with rat anti-mouse IL-6 receptor monoclonal antibody inhibited the increase in cathepsin activities in the injected muscle. We conclude that IL-6 produced in the inflamed muscle is involved in the process of muscle degeneration, especially through the activation of lysosomal cathepsins. PMID- 9348093 TI - Human cDNA encoding a novel TGF-beta superfamily protein highly expressed in placenta. AB - Recently, we developed a simple method for detecting a secretory signal sequence encoded by a cDNA fragment. In this study, we used this method to select cDNA clones encoding secretory proteins from a human full-length cDNA library. Full sequencing analysis of the candidate clones revealed that one clone encoded a novel TGF-beta superfamily protein. The clone encodes a protein of 308 amino acids of which the C-terminal region shows a characteristic feature of TGF-beta superfamily proteins: seven conserved cysteine residues at the C-terminal preceded by a putative processing site composed of a basic amino acid repeat. The corresponding transcripts are highly expressed in the placenta, so the novel protein may play an important role in reproduction. PMID- 9348092 TI - Expression of acetoacetyl-CoA thiolase isozyme genes of n-alkane-assimilating yeast, Candida tropicalis: isozymes in two intracellular compartments are derived from the same genes. AB - In the n-alkane-assimilating yeast Candida tropicalis, there are two isozymes of acetoacetyl-CoA thiolase, peroxisomal acetoacetyl-CoA thiolase (peroxisomal Thiolase I), and cytosolic acetoacetyl-CoA thiolase (cytosolic Thiolase I). We have previously isolated two genes (CT-T1A and CT-T1B) which encode Thiolase I. In order to compare the expressed products of Thiolase I isozyme-encoding genes in C. tropicalis, cytosolic Thiolase I was first purified from glucose-grown C. tropicalis in which the proliferation of peroxisomes and the expression of peroxisomal Thiolase I were repressed. Cytosolic Thiolase I was virtually identical to peroxisomal Thiolase I in molecular mass, kinetic and immunochemical properties, and primary structure at the N-terminus. Amino acid sequence analysis revealed that cytosolic Thiolase I was the mixture of products of two genes (CT T1A and CT-T1B), as in the case of the peroxisomal enzyme. CT-T1A and CT-T1B were expressed independently in the yeast Saccharomyces cerevisiae and the recombinant proteins were purified. Recombinant Thiolase IA and IB exhibited practically identical enzymatic properties to cytosolic and peroxisomal Thiolase Is from C. tropicalis. These results revealed that cytosolic Thiolase I and peroxisomal Thiolase I were encoded not by different genes, but by the same genes (CT-T1A and CT-T1B) and are present as a mixture of products expressed by both genes, although their subcellular localizations are different. PMID- 9348095 TI - Kinetic and regulatory properties of rat liver phosphoribosylpyrophosphate synthetase complex are partly distinct from those of isolated recombinant component catalytic subunits. AB - Rat liver phosphoribosylpyrophosphate (PRPP) synthetase exists as complex aggregates composed of two catalytic subunits (PRS I and II, in a ratio of approximately 4:1) and two catalytically inactive PRPP synthetase-associated proteins. To better understand the significance of the complex structure, the properties of the native liver enzyme were compared with those of homologous aggregates of recombinant PRS I and PRS II (rPRS I and rPRS II). (1) The specific activity per catalytic subunits of the liver enzyme was about 2.5 times lower than that of rPRS I over a wide pH range. Km values for substrates and Ka values for Pi and Mg2+ of the three enzymes were similar. (2) Specific activity of the liver enzyme for the reverse reaction was about 2 times lower than those of rPRSs. Km values for substrates of the three enzymes were comparable. (3) The liver enzyme was more stable than were rPRSs when incubated at a high temperature or in the absence of stabilizing agents. (4) The liver enzyme was markedly less sensitive to inhibition by nucleotides compared to rPRS I. GDP at 1 mM inhibited the liver enzyme and rPRS I by 32 and 93%, respectively. This effect is not ascribable to molecular interaction between rPRS I and II, as reconstitution of the two did not alter the sensitivity to nucleotide inhibition. (5) Our observations suggest that complex aggregation states of the native enzyme not only suppress the activities but also stabilize the catalytic subunits and the associated proteins and remarkably reduce the sensitivity to inhibition by nucleotides. PMID- 9348094 TI - Identification of two nucleotide-binding domains on the PB1 subunit of influenza virus RNA polymerase. AB - Influenza virus RNA polymerase is a multifunctional and multisubunit enzyme consisting of three viral P proteins, PB1, PB2, and PA. We have previously shown that radioactive 8-azido GTP (8-N3 GTP) was photo-crosslinked specifically to the PB1 subunit. Here we confirmed the specific crosslinking of PB1 using oxidized GTP and further identified the GTP analogue binding domains after proteolytic cleavage of the crosslinked PB1 with V8 protease. The cleavage pattern of PB1 was determined by analysis of the amino-terminal proximal sequence of fragments generated in the presence of increasing concentrations of V8 protease. The GTP crosslinking was identified in three fragments: two adjacent fragments, P6 starting from residue 179 and Pllb starting from residue 298; and the third fragment, P11c, starting from residue 458. Thus, we propose that two GTP-binding sites exist in the PB1 subunit, i.e., the amino terminal-proximal site I located at the boundary between P6 and Pllb, and the carboxy terminal proximal site II on P11c fragment. The locations of GTP-binding sites I and II are close to those of sequence motif A and motif D, respectively, conserved among RNA-dependent RNA polymerases. Of the two fragments forming site I, the crosslinking of 8-N3 GTP is higher to P11b, while that of oxidized GTP is higher to P6, suggesting that the ribose and guanine moieties of GTP bound in this binding pocket face P6 and P11c, respectively. From the V8 concentration dependent change in proteolytic cleavage pattern, it is likely that the two GTP-binding sites on PB1 protein are located on structurally different domains. The existence of two GTP-binding sites is discussed in relation to the binding sites for substrates, primers, and products. PMID- 9348096 TI - Analysis of the distribution of Na+/H+ exchanger isoforms among the plasma membrane subfractions of bovine kidney cortex: reevaluation of methods for fractionating the brush-border and the basolateral membranes. AB - Distribution of the NHE1 and the NHE3 isoforms of Na+/H+ exchanger in the plasma membranes of bovine kidney cortex was analyzed. Fractionation of the plasma membranes by centrifugation on a Percoll density gradient resulted in clear separation of the basolateral membranes (BLM) from the brush-border membranes (BBM), with Na+,K+-ATPase and aminopeptidase M as their respective marker enzymes. Under these conditions, a 110 kDa protein cross-reactive with an anti NHE1 antibody was detected exclusively in the BLM fractions, while a 90 kDa protein cross-reactive with an anti-NHE3 antibody was detected in the BBM fractions. A conventional Mg2+-precipitation method for obtaining the BBM, which is adequate with rabbit kidney as a starting material, turned out to be inadequate with bovine kidney cortex, since a considerable amount of the 110-kDa NHE1 protein was detected in the bovine kidney BBM fraction prepared by this procedure, together with the 90-kDa NHE3 protein. Percoll density gradient centrifugation is thus strongly recommended for the fractionation of BBM and BLM of bovine kidney cortex. The bovine NHE1 isoform was shown to be unique in that it is far less sensitive to the inhibition by ethylisopropylamiloride than that of other species. PMID- 9348097 TI - Purification and characterization of two mevalonate pyrophosphate decarboxylases from rat liver: a novel molecular species of 37 kDa. AB - The biosynthesis of cholesterol is regulated mainly by HMG-CoA reductase, however, recent studies indicated the pivotal role of another enzyme in cholesterol homeostasis. A previous report showed a marked decrease in the activity of mevalonate pyrophosphate decarboxylase (MPD) in stroke-prone spontaneously hypertensive rats and its possible involvement in the pathogenesis of the disorder. In this study, we purified liver MPD from rats fed a diet containing cholestyramine and pravastatin (CP diet) using conventional chromatographic techniques. We obtained two electrophoretically homogeneous enzyme preparations; 45 and 37 kDa proteins with specific activities of 8.0 and 7.4 micromol/min/mg, respectively. The enzymes showed similar molecular weights of 90 kDa, as judged on gel permeation chromatography. A kinetic study indicated apparent Km values for mevalonate pyrophosphate and ATP of 22.7 microM and 0.71 mM, respectively, for the 45 kDa MPD, and 20.0 microM and 0.80 mM, respectively, for the 37 kDa MPD. Half maximum activities were observed at 1.5 mM and 1.1 mM Mg2+ for the 45 and 37 kDa MPDs, respectively. Both enzymes required ATP as a phosphate acceptor, and in addition Mg2+, Mn2+, and Co2+ were effective as divalent cations. The optimum pH for both enzymes was 7.0. The isoelectric points for the 45 and 37 kDa MPDs were 5.6 and 5.4, respectively. Polyclonal antiserum raised against the 45 kDa enzyme detected both the 45 and 37 kDa bands on immunoblots with CP diet-induced liver crude extract as an antigen. However, non induced liver contained the 45 kDa protein but not the 37 kDa protein. These results indicated that the CP diet induced a new species, 37 kDa, of MPD which is characteristically and immunologically very similar to the well-known 45 kDa MPD. PMID- 9348098 TI - Bile acid profiles in a peroxisomal D-3-hydroxyacyl-CoA dehydratase/D-3 hydroxyacyl-CoA dehydrogenase bifunctional protein deficiency. AB - Bile acid profiles in serum, urine and bile from an infant with a peroxisomal D-3 hydroxyacyl-CoA dehydratase/D-3-hydroxyacyl-CoA dehydrogenase bifunctional protein (D-bifunctional protein) deficiency were analyzed by means of gas-liquid chromatography, gas-liquid chromatography-mass spectrometry, and high-performance liquid chromatography. As in such several peroxisomal disorders as Zellweger syndrome, neonatal adrenoleukodystrophy, and infantile Refsum disease, the accumulation of C27-bile acid intermediates was also demonstrated in the infant with D-bifunctional protein deficiency, accounting for 74% of the total bile acids in serum, 59% in urine, and 35% in bile. In addition, the major constituents of the C27-bile acids were (24R,25R)- and (24R,25S) 3alpha,7alpha,12alpha,24-tetrahydroxy-5be ta-cholestanoic acids along with small amounts of their 24S counterparts. Since immunoreactive acyl-CoA oxidase, L bifunctional protein, and thiolase were all present in the liver, the impairment of the oxidative side-chain cleavage in bile acid biosynthesis is considered to be due to the defect of D-bifunctional protein. PMID- 9348100 TI - Non-hydrated state of the acyl phosphate group in the phosphorylated intermediate of (Na+,K+)-ATPase. AB - The position in the acyl phosphate linkage of the phosphorylated intermediate of (Na+, K+)-ATPase that is cleaved by N-methylhydroxylamine was compared with that of the model compound acetylphosphate. The products of the cleavage of the phosphoenzyme by methylhydroxylamine were the active enzyme and a N-P compound, not the inhibited enzyme and inorganic phosphate. This means that the bond cleaved by methylhydroxylamine was the O-P bond, not the C-O bond. In contrast, methylhydroxylamine did not cleave the O-P bond of acetylphosphate in solution, at pH values from 0.3 to 7.0, whether or not the phosphoryl group formed a complex with magnesium. Acetylphosphate and hydroxylamine formed acetohydroxamic acid. Therefore, the state of the acyl phosphate bond in the native phosphoenzyme and in acetylphosphate in solution was different, and the difference was not due to different dissociation states of their phosphoryl groups or the binding of magnesium to the phosphoenzyme. Molecular orbital calculations for acetylphosphate revealed that the phosphorus atom charge is more positive than the carbon atom, irrespective of the dissociation state of the phosphoryl group. Similarly, the overlapping electron population of the O-P bond is always smaller than that of the C-O bond. Thus, the electronic structure of the acyl phosphate linkage of acetylphosphate under vacuum supports the results obtained with the native phosphoenzyme, rather than those obtained with acetylphosphate in solution. The linkage in the active site of the phosphorylated intermediate of (Na+,K+)-ATPase appeared to be equivalent to the non-hydrated state of the model compound acetylphosphate. The phosphoenzyme with bound ouabain, or without a tightly bound divalent cation was insensitive to methylhydroxylamine. The native phosphoenzyme of (Ca2+)-ATPase was not susceptible to methylhydroxylamine. PMID- 9348099 TI - ATP-induced dynamic fluorescence changes of a N-[p-(2 benzimidazolyl)phenyl]maleimide probe at Cys241 in the alpha-chain of pig stomach H+,K+-ATPase. AB - H+,K+-ATPase preparations from pig stomach were modified with a sulfhydryl fluorescence reagent, N-[p-(2-benzimidazolyl)phenyl] maleimide (BIPM). The addition of ATP to the modified enzyme preparations in the presence of Mg2+ decreased the BIPM fluorescence but increased the Trp fluorescence. After exhaustion of ATP, the fluorescence intensities increased and decreased to the original levels, respectively. The results of stopped flow and rapid quenching experiments suggested that the decrease in BIPM fluorescence (36/s) was accompanied by binding of Mg2+ and ATP or phosphorylation (35 36/s) which was followed by slower increases in Trp fluorescence (24/s) and light scattering (20/s). Tosylphenylalanyl chloromethyl ketone-trypsin treatment of the modified preparations, which showed an about 1% decrease in BIPM fluorescence accompanying phosphorylation, gave one major fluorescent peptide peak on reverse-phase chromatography. Amino acid sequence analysis of the peptide revealed the following sequence, Ser-Pro-Glu-X-Thr-His-Glu-Ser-Pro-Leu-Glu-Thr-Arg. On comparison with the amino acid sequence deduced from cDNA from pig stomach [Maeda, M., Ishizaki, J., and Futai, M. (1988) Biochem. Biophys. Res. Commun. 157, 203-209], X was shown to correspond to Cys241 of the alpha-chain in H+,K+ ATPase. These data and others suggest that the decrease in BIPM fluorescence at Cys241 reflects some molecular event triggered by the binding of ATP with Mg2+ and/or phosphorylation, whereas the increases in the intrinsic Trp fluorescence and light scattering reflect one after phosphorylation. PMID- 9348101 TI - A novel brain-derived member of the epidermal growth factor family that interacts with ErbB3 and ErbB4. AB - A novel member of the epidermal growth factor (EGF) family, the neural- and thymus-derived activator for ErbB kinases (NTAK), has been purified and cloned. Five alternative spliced isoforms have been detected in the rat adrenal pheochromocytoma cell line, PC-12 cells. The rat NTAK alpha2a isoform exhibits 94% identity in its primary sequence with the human NTAK alpha isoform. In vivo, NTAK is only expressed in the brain of rat E11.5 embryos, and in the brain and thymus of adult rats. The soluble 46 kDa form binds directly to ErbB3 and B4, but not to ErbB1 or B2. NTAK, however, transactivates ErbB1 and B2 via heterodimerization with ErbB3 or B4. NTAK stimulates the differentiation of MDA MB-453 cells and competitively inhibits the binding of [125I]neuregulin to these cells. In addition to these neuregulin-like properties, NTAK exhibits limited structural homology to neuregulins in the immunoglobulin (Ig)-like, EGF-like, and hydrophobic domains. Thus, NTAK appears to be a new member of the EGF family displaying neuregulin properties. PMID- 9348102 TI - Granny's remedy explained at the molecular level: helenalin inhibits NF-kappaB. PMID- 9348103 TI - The role of amyloid in the pathogenesis of Alzheimer's disease. AB - Since the identification in 1984 of the amyloid beta protein (Abeta) as the major component of senile plaques and cerebrovascular amyloid in Alzheimer's disease (AD) brains, it is well accepted that the production of this protein is a crucial factor in the pathogenesis of AD. Abeta is produced by cleavage from the amyloid precursor protein (APP) and can form fibrils in vivo and in vitro. The formation of these fibrils is influenced by proteins that are found in association with Abeta-containing lesions in the AD brain. Several of these proteins arise by an inflammatory response of the brain to Abeta production. The distribution of different isoforms of Abeta, varying at the C-terminus of the peptide, varies among the Abeta-containing lesions in AD brains. Such variations may have consequences for the pathogenesis of AD because the various Abeta isoforms differ in their capacity to form fibrils, and they have different toxic effects on neurons and vascular cells, respectively. The experimental data indicate that the pathogenesis of senile plaques is different from the generation of cerebrovascular amyloidosis. Summarizing models for either type of AD pathology are presented. PMID- 9348104 TI - Helenalin, an anti-inflammatory sesquiterpene lactone from Arnica, selectively inhibits transcription factor NF-kappaB. AB - Alcoholic extracts prepared form Arnicae flos, the collective name for flowerheads from Arnica montana and A. chamissonis ssp. foliosa, are used therapeutically as anti-inflammatory remedies. The active ingredients mediating the pharmacological effect are mainly sesquiterpene lactones, such as helenalin, 11alpha,13-dihydrohelenalin, chamissonolid and their ester derivatives. While these compounds affect various cellular processes, current data do not fully explain how sesquiterpene lactones exert their anti-inflammatory effect. We show here that helenalin, and, to a much lesser degree, 11alpha,13-dihydrohelenalin and chamissonolid, inhibit activation of transcription factor NF-kappaB. This difference in efficacy, which correlates with the compounds' anti-inflammatory potency in vivo, may be explained by differences in structure and conformation. NF-kappaB, which resides in an inactive, cytoplasmic complex in unstimulated cells, is activated by phosphorylation and degradation of its inhibitory subunit, IkappaB. Helenalin inhibits NF-kappaB activation in response to four different stimuli in T-cells, B-cells and epithelial cells and abrogates kappaB-driven gene expression. This inhibition is selective, as the activity of four other transcription factors, Oct-1, TBP, Sp1 and STAT 5 was not affected. We show that inhibition is not due to a direct modification of the active NF-kappaB heterodimer. Rather, helenalin modifies the NF-kappaB/IkappaB complex, preventing the release of IkappaB. These data suggest a molecular mechanism for the anti inflammatory effect of sesquiterpene lactones, which differs from that of other nonsteroidal anti-inflammatory drugs (NSAIDs), indomethacin and acetyl salicylic acid. PMID- 9348105 TI - DNA replication and order of cell cycle events: a role for protein isoprenylation? AB - When the aya1+ gene is mutated, Schizosaccharomyces pombe cells become unable to react appropriately to a delay in DNA replication. Instead of stalling the cell cycle to allow completion of DNA synthesis, they proceed unperturbed towards mitosis and attempt to segregate the still unreplicated chromosomes. As a result, the genetic material segregates unevenly and the nuclei assume a mitotic catastrophe phenotype, characterized by torn chromosomes (cut), anucleated cells and scattered chromosomes. Interestingly, the aya1 phenotype can be suppressed by overexpression of either the catalytic subunit of S. pombe DNA polymerase alpha or of a novel protein called hur1 +p. The latter bears significant homology to the core of the human Rab escort protein, which belongs to a family of factors necessary to the post-translational isoprenylation of proteins like Ras, Rab and lamin B. When isoprenylation is chemically inhibited with R-limonene (a monoterpene derived from orange rind), wild type S. pombe cells become insensitive to an S phase delay, in a manner strongly reminiscent of aya1 mutants. Moreover, overexpression of hur1 +p in wild type cells rescues the failing checkpoint function. We propose that there is a strong correlation between the aya1 phenotype, S-M phase checkpoint function, and isoprenylation events in fission yeast. PMID- 9348106 TI - The recognition of methylated DNA by the GTP-dependent restriction endonuclease McrBC resides in the N-terminal domain of McrB. AB - McrBC is a GTP-dependent restriction endonuclease of E. coli K12, selectively directed against DNA containing modified cytosine residues. McrB, one of its components, is responsible for the binding and, together with McrC, for the cleavage of DNAs containing two 5'-Pu(m)C sites separated by 40-80 base pairs. Gel retardation assays with wild-type and mutant McrB reveal that (i) single 5' Pu(m)C sites in DNA can be sufficient to elicite binding by McrB. Binding to such substrates is, however, weak and strongly dependent on the sequence context of Pu(m)C sites. (ii) Strong DNA binding (K(ass) approximately 10(7)M[-1]) is dependent on the presence of at least two Pu(m)C sites, even if they are separated by less than 40 bp, and is modulated by the sequence context ( A(m)CCGGT- --> -A(m)CT(C/G)AGT- --> -AGG(m)CCT- --> -AAG(m)CTT-). (iii) DNA binding by McrB is accompanied by formation of distinct multiple complexes whose distribution is modulated by GTP. (iv) McrC, which cannot bind DNA by itself, moderately stimulates the DNA binding of McrB and converts McrB-DNA complexes to large aggregates. (v) Deletion of the C-terminal half of McrB, which harbors the three consensus sequences characteristic for guanine nucleotide binding proteins, leads to protein inactive in GTP binding and/or hydrolysis and in McrC-assisted DNA cleavage; the protein, however, remains fully competent in DNA binding. (vi) Mutations in McrB which lead to a reduction in GTP binding and/or hydrolysis can affect DNA binding, suggesting that the two activities are coupled in the full length protein. PMID- 9348107 TI - Determination of the regulatory disulfide bonds of NADP-dependent malate dehydrogenase from Pisum sativum by site-directed mutagenesis. AB - The light-mediated reversible activation of NADP-dependent malate dehydrogenase (NADP-MDH) from Pisum sativum can be simulated in vitro by reducing the inactive oxidized enzyme with dithiothreitol. Since the gross structure and the dimeric state of the enzyme are unaffected by the state of oxidation, the redox modulation cannot be attributed to inter-subunit disulfide bridges. In order to identify intra-chain cystine cross bridges that might be candidates responsible for the activation reaction, site-directed mutagenesis experiments were performed, substituting alanine for up to four exposed cysteine residues. Mutants were expressed in freshly transformed EcoB cells and purified to homogeneity. As indicated by the activation behavior (by dithiothreitol-mediated thioldisulfide exchange), disulfides C23-C28 in the N-terminal and C364-C376 in the C-terminal part of the polypeptide chain are involved in the light-induced modulation of the activity of the wild type enzyme. A mutant of the enzyme lacking the N-terminal 45 residues confirms this result. Electrophoretic mobility and FPLC prove the wild type enzyme and its mutants to be dimeric; differences refer to the packing of the N- and C-terminal portions of the enzyme in its oxidized and reduced state. The kinetics of the redox modulation differ, depending on the solvent conditions and the mode of activation. After elimination of the N-terminal disulfide bond, sigmoidal activation profiles are no longer observed, suggesting a slow conformational rearrangement in the N-terminal portion of the wild type enzyme to be rate-limiting in the course of reductive activation. For the wild type, this finding can be mimicked in the presence of non-denaturing concentrations of guanidinium-chloride. PMID- 9348108 TI - Amino acid sequence, S-S bridge arrangement and distribution in plant tissues of thionins from Viscum album. AB - The complete primary structure of a cytotoxic 5 kDa polypeptide, viscotoxin A1, isolated from Viscum album L., has been determined by combining classical Edman degradation methodology with advanced mass spectrometric procedures. The same integrated approach allowed correction of the sequence of viscotoxin A2 and definition of the pattern of the disulfide bridges. The arrangement of the cysteine pairing was determined as Cys3-Cys40, Cys4-Cys32 and Cys16-Cys26. The primary structure of viscotoxin A1 shares a high degree of similarity with the known viscotoxins and more generally with the plant alpha- and beta-thionins. The pattern of S-S bridges determined for viscotoxin A2 and A1 is similar to that inferred by X-ray and NMR analysis in crambin and related to that present in alpha-purothionin and beta-hordothionin, thus indicating a highly conserved organization of the S-S pairings within the entire family. This arrangement of S S bridges describes a peculiar structural motif, indicated as 'concentric motif', which is suggested to stabilize a common structure occurring in various small proteins able to interact with cell membranes. The distribution of the new variant toxin in different mistletoe subspecies was investigated. Viscotoxin A1 is abundant in the seeds of the three European subspecies of V. album whereas it represents a minor component in the shoots. PMID- 9348109 TI - Characterization of monoclonal and polyclonal antibodies to human choline acetyltransferase and epitope analysis. AB - Choline acetyltransferase (ChAT) was partially purified from human placenta and brain. In order to raise monoclonal antibodies, Balb/c mice were immunized with a preparation from placenta or with a mixture of eight synthetic peptides that were deduced from the primary structures of porcine and human ChAT. Polyclonal antibodies were raised in rabbits against five synthetic peptides deduced from the amino acid sequence of human ChAT. The monoclonal and polyclonal antibodies were characterized by their ability to recognize ChAT in various immunoassays: immunoblot, enzyme-linked immunosorbent assay (ELISA), two-side ELISA and immunohistochemistry. With one exception all monoclonal antibodies recognized ChAT on immunoblots, some were particularly sensitive; one bound active ChAT in ELISA when used as capture reagent; most antibodies recognized immobilized ChAT in ELISA. Two monoclonal antibodies out of nine gave particularly excellent results in staining cholinergic neurons and fibers on sections from rat and primate brain. With the help of nine synthetic peptides it was possible to evaluate two major binding sites for the monoclonal antibodies on the ChAT molecule, comprising amino acids 167-189 and 57-76, respectively. PMID- 9348110 TI - Full length cDNA of rat RT1.DMa and RT1.DMb and expression of RT1.DM genes in dendritic and Langerhans cells. AB - MHC encoded DM heterodimers and classical MHC class II complexes meet in an endosomal/lysosomal compartment where DM heterodimers support peptide loading of MHC class II. Studies on peptide loading of rat class II and on peptide persistence in cells of the dendritic lineage prompted us to establish full length cDNA clones coding for the subunits alpha and beta of rat DM molecules as well as a mAb directed against the luminal moiety of the beta subunit. Here we describe the establishment of the first full length cDNA clones of rat RT1.DMa and RT1.DMb. The mode of expression of RT1.DM at the transcript level in bone marrow culture-derived dendritic cells, in Langerhans cells and in a number of additional accessory cells is reported. The beta protein was identified in detergent lysates of RT1.DM expressing cells by Western blot analysis using a newly established monoclonal antibody directed against the luminal part of RT1.DMbeta. PMID- 9348111 TI - ATP and ADP bind to cytochrome c oxidase and regulate its activity. AB - By equilibrium dialysis of cytochrome c oxidase from bovine heart with [35S]ATPalphaS and [35S]ADPalphaS, seven binding sites for ATP and ten for ADP were determined per monomer of the isolated enzyme. The binding of ATP occurs in a time-dependent manner, as shown by a filtration method, which is apparently due to slow exchange of bound cholate. In the crystallized enzyme 10 mol of cholate were determined and partly identified in the high resolution crystal structure. Binding of ADP leads to conformational changes of the Tween 20-solubilized enzyme, as shown by a 12% decrease of the gamma-band. The conformational change is specific for ADP, since CDP, GDP and UDP showed no effects. The spectral changes are not obtained with the dodecylmaltoside solubilized enzyme. The polarographically measured activity of cytochrome c oxidase is lower after preincubation with high ATP/ADP-ratios than with low, in the presence of Tween 20. This effect of nucleotides is due to interaction with subunit IV, because preincubation of the enzyme with a monoclonal antibody to subunit IV released the inhibition by ATP. In the presence of dodecylmaltoside the enzyme had a 2 to 3 fold higher total activity, but this activity was not influenced by preincubation with ATP or ADP. PMID- 9348112 TI - Selective modulation of protein kinase A and protein kinase C activities in epidermal growth factor (EGF)-stimulated MCF-7 breast cancer cells. AB - In human MCF-7 breast cancer cells, both protein kinase A (PKA) and different members of the protein kinase C (PKC) family are stimulated upon binding of epidermal growth factor (EGF) to cell surface receptors. Selective stimulation of calcium-dependent PKCs with 10(-6) to 10(-9) M Thymeleatoxin significantly increased the proliferation rate of MCF-7 cells over 5 days in culture. This stimulation was blocked by the PKC antagonist Chelerythrine. In contrast, selective activation of PKA by addition of 1 mM dibutyryl cyclic AMP (dBcAMP) did not affect the proliferation rate of MCF-7 cells. Similarly, activation of the adenylate cyclase by 1 microM Forskolin and inhibition of PKA by the cyclic AMP analogue Rp-cAMPS did not modulate the proliferation rate of these cells. Activation of PKC stimulated the expression of the immediate early gene c-fos but c-myc expression was not significantly enhanced. On the other hand, PKA activation increased both c-myc and c-fos expression in MCF-7 cells. These results suggest that PKA activation and c-myc expression are not obligatory for proliferation of MCF-7 cells. PMID- 9348113 TI - Affinity purification and characterization of glucosidase II from pig liver. AB - Glucosidase II has been purified from crude pig liver microsomes by a convenient procedure involving DEAE-Sephacel, Con A-Sepharose and affinity chromatography on N-5-carboxypentyl-1-deoxynojirimycin-AH-Sepharose. Specific binding of glucosidase II to the affinity matrix required its prior separation from glucosidase I, which was accomplished by fractional Con A-Sepharose chromatography. The three-step procedure yielded, with approximately 15% enzyme recovery, a > 190-fold enriched glucosidase II, consisting of two proteins (107 kDa and 112 kDa). Both polypeptides are N-glycosylated with probably one glycan chain, in line with their binding to Con A-Sepharose. Immunological cross reactivity and other experimental data indicate that the 107 kDa N-glycoprotein is derived from the 112 kDa species by partial proteolysis. The occasional presence of a 60 kDa peptide co-eluting with the catalytic activity suggests that glucosidase II may be associated with other protein subunit(s) in a heteromeric membrane complex. Glucosidase II hydrolyzes the alpha1,3-glucosidic linkages in Glc(2-1)-Man9-GlcNAc2, as well as synthetic alpha-glucosides, efficiently but does not remove the distal alpha1,2-linked glucose in Glc3-Man9-GlcNAc2. The enzyme has a pH optimum close to 6.5 and is not metal ion-dependent. Catalytic activity is strongly inhibited by basic sugar analogues including 1 deoxynojirimycin (dNM; app. Ki approximately 7.0 microM), N-5-carboxypentyl-dNM (app. Ki approximately 32 microM) and castanospermine (app. Ki approximately 40 microM). Substitution of the 3-OH or 6-OH group in dNM by a fluoro group reduces the inhibitory potential drastically. We conclude from these observations that the two hydroxy groups are essential for inhibitor/substrate binding due to their ability to interfere as hydrogen bond donors. A polyclonal antibody raised against the 107 kDa polypeptide reacted specifically with two proteins from different cell types on Western blots. Their molecular masses were identical with those from pig liver microsomes, pointing to a highly conserved amino acid sequence of glucosidase II. This suggests that the variance in molecular mass for glucosidase II reported for the enzyme from other tissues and species may be due to partial proteolysis. PMID- 9348114 TI - Regulation of the intracellular pH in the protozoan parasite Trypanosoma brucei brucei. AB - The mechanisms regulating the intracellular pH (pHi) in both forms of Trypanosoma brucei brucei (cultured cells) were investigated using the fluorescent probe 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF). The pHi values measured were 7.22+/-0.03 in the procyclics and 7.40+/-0.05 in the bloodstream form. In the presence of 24mM HCO3-, pHi values were slightly higher in both forms of trypanosomes suggesting a bicarbonate-linked pH regulation. pHi was more stable in procyclics (between 7.15 and 7.30 in the external pH range 6.4-7.6) than in the bloodstream forms. The amiloride analogue tested decreased pHi, suggesting Na+-driven Na+/H+ antiporters. H+-ATPases also seem to be involved in pHi regulation since the inhibitors N-ethylmaleimide (1 mM) and N,N' dicyclohexylcarbodiimide (80 microM) induced a rapid acidification in both forms of trypanosomes. Addition of pyruvate caused a cytosol acidification in the bloodstream form only confirming the existence of a diffusion-facilitated carrier for pyruvate, with the cotransport of H+. Our results show that, although similar pH regulation mechanisms seem to exist in both forms of trypanosomes, the procyclics can regulate efficiently their pHi and consequently their plasma membrane potential whereas the bloodstream forms cannot always maintain their pHi and are easily depolarized following a small acid load. PMID- 9348115 TI - Subcellular localization of dihydropyrimidine dehydrogenase. AB - Although dihydropyrimidine dehydrogenase (DPD) has been purified and characterized from liver tissues of various mammals conflicting data exist on its subcellular localization. To determine the localization of DPD we prepared crude subcellular fractions of a rat liver homogenate by means of differential centrifugation. In the fractions obtained (heavy mitochondrial, light mitochondrial, microsomal and cytosolic) the activities of different marker enzymes were measured as well as the activity of DPD. These results showed that almost all of the activity of DPD was located in the cytosolic fraction. To exclude any particulate-associated DPD, a light mitochondrial fraction was subsequently subjected to equilibrium density gradient centrifugation. The distribution profile of the activity of DPD and the various marker enzymes indicated that DPD from rat liver was exclusively located in the cytosol since no significant activity of DPD could be detected in any subcellular organelle. PMID- 9348116 TI - Distribution of the secretory leucocyte proteinase inhibitor in human articular cartilage. AB - The secretory leucocyte protease inhibitor, SLPI, is a low molecular weight inhibitor of proteases such as elastase and cathepsin G, which are released from leucocytes during phagocytosis. The purpose of this study was to show whether or not SLPI is produced in articular chondrocytes. In articular disorders, the protease-antiprotease balance is disturbed. For this reason it would be interesting to establish the source of SLPI. The presence of SLPI was demonstrated using immunohistochemistry and in situ hybridization, hence we conclude that SLPI is produced in the chondrocytes of human articular cartilage. PMID- 9348117 TI - Mechanisms of antithrombin polymerisation and heparin activation probed by the insertion of synthetic reactive loop peptides. AB - Incubation of antithrombin with a series of synthetic reactive loop peptides showed that 6-mer and 7-mer peptides, P14-P9 and P14-P8 of antithrombin respectively, induced loop-sheet polymerisation and binary complex formation. These peptides are likely to anneal to the upper part of the dominant A-sheet, favouring sheet opening and allowing insertion of a second reactive loop in the lower part of the A-sheet to form polymers. The insertion of longer peptides filled the A-sheet beyond the P7 position and prevented polymerisation. Heparinised antithrombin was more resistant to polymerisation and peptide insertion, indicating that heparin induces a conformational change that closes the A-sheet and expels the reactive loop. PMID- 9348118 TI - Dependence on intracellular Ca2+ on mass and turnover of phosphoinositides and phosphatidate in human erythrocytes. AB - Effects of a calcium-load on mass and turnover of phosphoinositides and phosphatidate were investigated in human erythrocytes by short-term labeling with [32P]Pi. The labeling of phosphatidate was accelerated at normal mass by short term elevation of free intracellular [Ca2+] up to 1 microM and inhibited by the reduction of normal free [Ca2+]. Thus, the labeling of phosphatidate is a Ca2+ regulated process and not only the consequence of a net synthesis of diacylglycerol by other Ca2+-dependent reactions. Persisting elevation of free intracellular [Ca2+] between 1-40 microM induced an increase of the mass of phosphatidylinositol 4-phosphate with a concomitant decrease of the mass of phosphatidylinositol 4,5-bisphosphate. Under these conditions, the normal steady state turnover of phosphoinositides was not altered by Ca2+, but mass and turnover of phosphatidate continuously rose. The increase in phosphatidate mass by far exceeded the decrease of the mass of phosphoinositides, indicating that phosphatidate was generated to a great extent by hydrolysis of other phospholipids in addition to the action of phosphoinositidase C with subsequent phosphorylation of diacylglycerol to phosphatidate. The results demonstrate that different phospholipid phosphodiesterases of human erythrocytes are activated by Ca2+-concentrations in the microM range as is known from various other cell types. In contrast to current explanations, Ca2+-dependent phospholipid phosphodiesterases of human erythrocytes did not exhibit an unusually low affinity against rising cytosolic Ca2+-concentrations. PMID- 9348119 TI - The reaction of peroxynitrite with tert-butyl hydroperoxide produces singlet molecular oxygen. AB - Peroxynitrite, a biological oxidant, can induce lipid peroxidation in biological membranes which leads to the formation of various hydroperoxides. Here, we report the formation of singlet oxygen (1O2) in the reaction of peroxynitrite with tert butyl hydroperoxide (t-BOOH) used as a model compound for organic hydroperoxides. The formation of singlet oxygen was observed by (i) dimol emission in the red spectral region, (ii) monomol emission in the infrared region at 1270 nm and by (iii) chemical trapping of singlet oxygen with anthracene-9,10-diyldiethyl disulfate (EAS). The emission signal was increased when H2O was replaced by deuterium oxide and was quenched by sodium azide. When singlet oxygen was generated in the reaction of peroxynitrite with t-BOOH, the 1O2 quenching rate constant for sodium azide was estimated from a Stern-Volmer plot as 1.3 x 10(8) M(-1) S(-1) which is in line with known values. The 1O2 generation in the peroxynitrite/t-BOOH reaction was also detected by the formation of the endoperoxide of EAS. These data establish the generation of 1O2 in the reaction of peroxynitrite with t-BOOH and suggest a potential involvement of 1O2 in peroxynitrite-mediated reactions in biological systems. PMID- 9348120 TI - Effects of stimulus conditions on the performance of antisaccades in man. AB - We investigated the effect of different spatial and temporal parameters on the saccadic reaction times (SRTs) of the antisaccades and on the frequency and the SRTs of erratic prosaccades in five adult human subjects. The subjects were instructed to aim their saccades to the side opposite to where a visual go stimulus occurred. Parameters under consideration were: the gap duration (between 0 and 600 ms, and an overlap paradigm); the stimulus size (sizes of 0.1 degrees, 0.2 degrees, and 0.4 degrees, using the gap 200-ms paradigm); and the stimulus eccentricity (1 degree, 2 degrees, 4 degrees, 8 degrees, and 12 degrees, with the gap 200-ms paradigm). A decrease in the anti SRTs and an increase in the error rate were observed with medium gap durations (200 ms, 250 ms), while the anti SRTs were longer and the error rates lower with the shorter values (0 ms, 100 ms, and with the overlap paradigm) and with the long values (600 ms). A slight decrease in the anti-SRTs and an increase in the error frequency occurred with increasing eccentricity; the SRT distributions of the errors resembled closely those of prosaccades in corresponding prosaccade tasks with the same eccentricities. The stimulus size had only modest or no effects at all. Analysis of the distributions of the correction times of the erratic prosaccades showed that the intersaccadic intervals could be very short: in the range of express saccades, with a peak at 100 ms or in some subjects even shorter, with a peak at 40-50 ms. It is concluded that the performance of antisaccades is influenced by parameters that interact with the fixation and/or attention system of oculomotor control. Parameters supporting a disengagement of fixation at the time of stimulus onset provoke a reduction of the saccadic reaction times not only of prosaccades but also of antisaccades. Moreover, a certain state of disengagement seems to facilitate the occurrence of reflex-like errors. PMID- 9348121 TI - Peak velocities of visually and nonvisually guided saccades in smooth-pursuit and saccadic tasks. AB - Smooth pursuit typically includes corrective catch-up saccades, but may also include such intrusive saccades away from the target as anticipatory or large overshooting saccades. We sought to differentiate catch-up from anticipatory and overshooting saccades by their peak velocities, to see whether the higher velocities of visually rather than nonvisually guided saccades in saccadic tasks may be found also in saccades in pursuit. In experiment 1, 12 subjects showed catch-up, anticipatory, and overshooting saccades to comprise 70.4% of all saccades in pursuit of periodic, 30 degrees/s constant-velocity targets. Catch-up saccades were faster than the others. Saccadic tasks were run as well, on 19 subjects, including the 12 whose pursuit data were analyzed, with target-onset, target-remaining (saccade to the remaining target when the other three extinguish), and antisaccade tasks. For 17 of the 19 subjects, antisaccade velocities were lower than for either target-onset or target-remaining tasks. Velocities for the target-remaining task were near those for target onset, indicating that target presence, not its onset, defines visually guided saccades. Error and reaction-time data suggest greater cognitive difficulty for target remaining than for target onset, so that the cognitive difficulty of typical nonvisually guided saccade tasks is not sufficient to produce their lowered velocity. To produce reliably, in each subject, catch-up and anticipatory saccades with comparable amplitude distributions, nine new subjects were asked in experiment 2 to make intentional catch-up and anticipatory saccades in pursuit, and were presented with embedded target jumps to elicit catch-up saccades, all with periodic target trajectories of 15 degrees/s and 30 degrees/s. Velocities of intentional anticipatory saccades were lower than velocities of intentional catch up saccades, while velocities of intentional and embedded catch-up saccades were similar. Target-onset and remembered-target saccadic tasks were run, showing the expected higher velocity for the target-onset task in each subject. Both experiments demonstrate higher peak velocities for catch-up saccades than for anticipatory saccades, suggesting that cortical structures preferentially involved in nonvisually guided saccades may initiate the anticipatory and overshooting saccades in pursuit. PMID- 9348122 TI - Characteristics of surround inhibition in cat area 17. AB - The effects of stimuli falling outside the 'classical receptive field' and their influence on the orientation selectivity of cells in the cat primary visual cortex are still matters of debate. Here we examine the variety of effects of such peripheral stimuli on responses to stimuli limited to the receptive field. We first determined the extent of the classical receptive field by increasing the diameter of a circular patch of drifting grating until the response saturated or reached a maximum, and by decreasing the diameter of a circular mask in the middle of an extended grating, centred on the receptive field, until the cell just began to respond. These two estimates always agreed closely. We then presented an optimum grating of medium-to-high contrast filling the classical receptive field while stimulating the surround with a drifting grating that had the same parameters as the central stimulus but was varied in orientation. For all but five neurons (of 37 tested), surround stimulation produced clear suppression over some range of orientations, while none showed explicit facilitation under these conditions. For 11 cells (34% of those showing suppression), the magnitude of suppression did not vary consistently with the orientation of the surround stimulus. In the majority of cells, suppression was weakest for a surround grating oriented orthogonal to the cell's optimum. Nine of these cells (28%) exhibited maximum inhibition at the optimum orientation for the receptive field itself, but for 12 cells (38%) there was apparent 'release' from inhibition for surround gratings at or near the cell's optimum orientation and direction, leaving inhibition either maximal at angles flanking the optimum (9 cells) or broadly distributed over the rest of the orientation range (3 cells). This implies the existence of a subliminal facilitatory mechanism, tightly tuned at or near the cell's optimum orientation, extending outside the classical receptive field. For just two cells of 13 tested the preferred orientation for a central grating was clearly shifted towards the orientation of a surrounding grating tilted away from the cell's optimum. The contrast gain for central stimulation at the optimal orientation was measured with and without a surround pattern. For nine of 25 cells tested, surround stimulation at the cell's optimum orientation facilitated the response to a central grating of low contrast (< or =0.1) but inhibited that to a higher-contrast central stimulus: the contrast response gain is reduced but the threshold contrast is actually decreased by surround stimulation. Hence the receptive field is effectively larger for low contrast than for high-contrast stimuli. Inhibition from the periphery is usually greatest at or around the cell's optimum, while suppression within the receptive field has been shown to be largely non-selective for orientation. Inhibition by orientations flanking the optimum could serve to sharpen orientation selectivity in the presence of contextual stimuli and to enhance orientational contrast; and it may play a part in orientation contrast illusions. PMID- 9348123 TI - Reversible inactivation of macaque frontal eye field. AB - The macaque frontal eye field (FEF) is involved in the generation of saccadic eye movements and fixations. To better understand the role of the FEF, we reversibly inactivated a portion of it while a monkey made saccades and fixations in response to visual stimuli. Lidocaine was infused into a FEF and neural inactivation was monitored with a nearby microelectrode. We used two saccadic tasks. In the delay task, a target was presented and then extinguished, but the monkey was not allowed to make a saccade to its location until a cue to move was given. In the step task, the monkey was allowed to look at a target as soon as it appeared. During FEF inactivation, monkeys were severely impaired at making saccades to locations of extinguished contralateral targets in the delay task. They were similarly impaired at making saccades to locations of contralateral targets in the step task if the target was flashed for < or =100 ms, such that it was gone before the saccade was initiated. Deficits included increases in saccadic latency, increases in saccadic error, and increases in the frequency of trials in which a saccade was not made. We varied the initial fixation location and found that the impairment specifically affected contraversive saccades rather than affecting all saccades made into head-centered contralateral space. Monkeys were impaired only slightly at making saccades to contralateral targets in the step task if the target duration was 1000 ms, such that the target was present during the saccade: latency increased, but increases in saccadic error were mild and increases in the frequency of trials in which a saccade was not made were insignificant. During FEF inactivation there usually was a direct correlation between the latency and the error of saccades made in response to contralateral targets. In the delay task, FEF inactivation increased the frequency of making premature saccades to ipsilateral targets. FEF inactivation had inconsistent and mild effects on saccadic peak velocity. FEF inactivation caused impairments in the ability to fixate lights steadily in contralateral space. FEF inactivation always caused an ipsiversive deviation of the eyes in darkness. In summary, our results suggest that the FEF plays major roles in (1) generating contraversive saccades to locations of extinguished or flashed targets, (2) maintaining contralateral fixations, and (3) suppressing inappropriate ipsiversive saccades. PMID- 9348124 TI - Organization of the entorhinal projection to the rat dentate gyrus revealed by Dil anterograde labeling. AB - It has been suggested that the entorhino-hippocampal circuit is involved in memory formation. To investigate the way that associative memory is elaborated in the circuit, the entorhino-dentate projection was studied with the fluorescent lipophilic tracer Dil. We investigated the projection originating in the dorsal part of the entorhinal cortex by injecting Dil along the rhinal sulcus. Anterograde fluorescent labeling allowed us to examine sections of the sample with a confocal microscope or in wholemount preparations with a fluorescence microscope. Quantitative analysis of the distribution of the Dil-labeled perforant path by confocal microscopy was performed in the septal one third level of the hippocampus. The analysis confirmed that the topographical map along the mediolateral dimension of the entorhinal cortex was transferred to the proximodistal level (from the inner one third to the edge of the molecular layer) of the granule cell dendrites in a gradually shifting manner. The fiber profile observed after lateral entorhinal injection was thick in the suprapyramidal blade and thin in the infrapyramidal blade. The fiber profile observed after medial entorhinal injection was thin in the suprapyramidal blade and thick in the infrapyramidal blade. Fluorescence microscopic observation of wholemount preparations showed that projections from the Dil injection site were distributed wider than half the dentate gyrus in the longitudinal direction. In transverse sections, the range of the labeled fiber distribution was confirmed to be more than two thirds of the dentate gyrus in the same direction regardless of the mediolateral level of the injection site. It has been suggested that the dorsoventral axis of the entorhinal cortex is represented in the septotemporal levels of the dentate gyrus, but that the topographical correspondence might be weak and vague. Although our investigation was limited to the projection from the dorsal entorhinal cortex to the dorsal part of the dentate gyrus, we conclude that the widely distributed projection covers the dentate gyrus in a nontopographic manner. PMID- 9348125 TI - Bioelectrical cochlear noise and its contralateral suppression: relation to background activity of the eighth nerve and effects of sedation and anesthesia. AB - The bioelectrical activity of the cochlea, without any ipsilateral acoustic stimulation, was recorded in awake guinea pigs (GPs) between electrodes chronically implanted at the round window (RW) and the skull. Measuring its power in the band centered around 1.0 kHz (0.5-2.5 kHz) provided an indirect measure of the ensemble background (EBA) activity of the eighth nerve. Contralateral white noise (CLWN) stimulation reduced this EBA, presumably by activation of medial olivocochlear fibers. The aim of the investigation was to validate measurements of EBA and of its contralateral suppression in order to study the medial efferent function. The first goal was to find the best conditions for recording the EBA in the absence of ipsilateral stimulation and for studying its suppression by contralateral acoustic stimulation, which implies that no noise was generated by the experimental animal. Thus recordings were compared in normal, awake GPs and in GPs under sedation with xylazine, anesthetized with a combination of xylazine and ketamine, and with and without temperature regulation. In order to monitor the effects of sedation and anesthesia, the recordings were analyzed not only in the 0.5- to 2.5-kHz frequency band but also in the other frequency bands, 5-50 Hz, 50-150 Hz, and 150-500 Hz, which presumably include general central and neuromuscular contributions. The results show that sedation with xylazine accompanied by regulation of body temperature does not affect the EBA value nor its contralateral suppression. Nevertheless, anesthesia should be avoided, even with control of body temperature. The second goal of this study was to identify the specific cochlear contribution to the raw RW signal. Thus recordings were performed in normal and deafened animals and analyzed in the frequency band 0.5 2.5 kHz and also in the other frequency bands of 5-50 Hz, 50-150 Hz, and 150-500 Hz. The results indicate that most of the cochlear activity lies in the frequency band 0.5-2.5 kHz, with also some minor contribution coming from the 150- to 500 Hz band. Analysis and comparison of power values in the different conditions indicate that specific cochlear EBA power was about 60 microV2. From a commonly accepted mean background discharge rate of 50 spikes/s (sp/s), the EBA power without CLWN should have been around 4.4 microV2 if the fibers' activity was random. This difference suggests that there is probably some degree of synchrony between individual fibers. There was a reduction of approximately 45% during CLWN stimulation. This suppression might correspond to a reduction in both discharge rate and synchrony of the fibers. PMID- 9348126 TI - Connections of the caudal ventrolateral medullary reticular formation in the cat brainstem. AB - A region of the caudal ventrolateral medullary reticular formation (CVLM) participates in baroreceptor, vestibulosympathetic, and somatosympathetic reflexes; the adjacent retroambigual area is involved in generating respiratory related activity and is essential for control of the upper airway during vocalization. However, little is known about the connections of the CVLM in the cat. In order to determine the locations of terminations of CVLM neurons, the anterograde tracers Phaseolus vulgaris leucoagglutinin and tetramethylrhodamine dextran amine were injected into this region. These injections produced a dense concentration of labeled axons throughout the lateral medullary reticular formation (lateral tegmental field), including the retrofacial nucleus and nucleus ambiguus, regions of the rostral ventrolateral medulla, the lateral and ventrolateral aspects of the hypoglossal nucleus, nucleus intercalatus, and the facial nucleus. A smaller number of labeled axons were located in the medial, lateral, and commissural subnuclei of nucleus tractus solitarius, the A5 region of the pontine reticular formation, the ventral and medial portions of the spinal and motor trigeminal nuclei, locus coeruleus, and the parabrachial nucleus. We confirmed the projection from the CVLM to both the rostral ventrolateral medulla and lateral tegmental field using retrograde tracing. Injections of biotinylated dextran amine or Fluorogold into these regions resulted in retrogradely labeled cell bodies in the CVLM. However, the neurons projecting to the lateral tegmental field were located mainly dorsal to those projecting to the rostral ventrolateral medulla, suggesting that these neurons form two groups, possibly with different inputs. Injections of retrograde tracers into the lateral tegmental field and rostral ventrolateral medulla also produced labeled cell bodies in other regions, including the medial and inferior vestibular nuclei and nucleus solitarius. These data are consistent with the view that the CVLM of the cat is a multifunctional area that regulates blood pressure, produces vocalization, affects the shape of the oral cavity, and elicits contraction of particular facial muscles. PMID- 9348127 TI - Segmental and propriospinal projection systems of frog lumbar interneurons. AB - Spinal interneuronal networks have been implicated in the coordination of reflex behaviors and limb postures in the spinal frog. As a first step in defining these networks, retrograde transport of horseradish peroxidase (HRP) was used to examine the anatomical organization of interneuronal circuitry in the lumbar spinal cord of the frog. Following neuronal degeneration induced by spinal transection and section of the dorsal and ventral roots, HRP was placed at different locations in the spinal cord and the positions of labeled neuronal cell bodies plotted using a Eutectics Neuron Tracing System. We describe four spinal interneuronal systems, three with cell bodies located in the lumbar cord and one with descending projections to the lumbar cord. Interneurons with cell bodies located in the lumbar cord include: (1) Lumbar neurons projecting rostrally. Those projecting to thoracic segments tended to be located in the lateral and ventrolateral gray and in the lower two-thirds of the dorsal horn, with projections that were predominantly uncrossed. Those projecting to the brachial plexus and beyond were located in the dorsal part of the dorsal horn (uncrossed) and in the lateral, ventrolateral, and ventromedial gray (crossed). (2) Lumbar neurons with segmental projections within the lumbar cord. These neurons, which were by far the most numerous, had both uncrossed and crossed projections and were distributed throughout the dorsal, lateral, ventrolateral, and ventromedial gray matter. (3) Lumbar neurons projecting to the sacral cord. This population, which arose mainly from the dorsal horn and lateral or ventrolateral gray, was much smaller than in the other systems. Neuronal density of some of these populations of lumbar interneurons appeared to vary with rostrocaudal level. Finally, a population of neurons with cell bodies in the brachial and thoracic segments that projects to the lumbar cord is described. The most rostral of these neurons were multipolar cells with uncrossed projections, while those with crossed projections were confined almost exclusively to the ventral half of the cord. The distribution of spinal interneurons reported here will provide guidance for future studies of the role of interneuronal networks in the control of movements using the spinal frog as a model system. PMID- 9348128 TI - Phasic and tonic stretch reflexes in muscles with few muscle spindles: human jaw opener muscles. AB - We investigated phasic and tonic stretch reflexes in human jaw-opener muscles, which have few, if any, muscle spindles. Jaw-unloading reflexes were recorded for both opener and closer muscles. Surface electromyographic (EMG) activity was obtained from left and right digastric and superficial masseter muscles, and jaw orientation and torques were recorded. Unloading of jaw-opener muscles elicited a short-latency decrease in EMG activity (averaging 20 ms) followed by a short duration silent period in these muscles and sometimes a short burst of activity in their antagonists. Similar behavior in response to unloading was observed for spindle-rich jaw-closer muscles, although the latency of the silent period was statistically shorter than that observed for jaw-opener muscles (averaging 13 ms). Control studies suggest that the jaw-opener reflex was not due to inputs from either cutaneous or periodontal mechanoreceptors. In the unloading response of the jaw openers, the tonic level of EMG activity observed after transition to the new jaw orientation was monotonically related to the residual torque and orientation. This is consistent with the idea that the tonic stretch reflex might mediate the change in muscle activation. In addition, the values of the static net joint torque and jaw orientation after the dynamic phase of unloading were related by a monotonic function resembling the invariant characteristic recorded in human limb joints. The torque-angle characteristics associated with different initial jaw orientations were similar in shape but spatially shifted, consistent with the idea that voluntary changes in jaw orientation might be associated with a change in a single parameter, which might be identified as the threshold of the tonic stretch reflex. It is suggested that functionally significant phasic and tonic stretch reflexes might not be mediated exclusively by muscle spindle afferents. Thus, the hypothesis that central modifications in the threshold of the tonic stretch reflex underlie the control of movement may be applied to the jaw system. PMID- 9348129 TI - Regional cerebral blood flow increases during preparation for and processing of sensory stimuli. AB - Preparing for and processing of sensory stimuli are energy-requiring processes. We attempted to assess the relative contributions of these processes to increases in regional cerebral perfusion. Nineteen healthy right-handed subjects were examined while they were engaged in detecting tactile stimuli to the index finger 5 s after a cueing tone. Cerebral blood flow velocity (CBFV) modulations in the middle cerebral arteries (MCAs) were continuously measured by bilateral simultaneous transcranial Doppler ultrasonography. Tactile stimuli well above threshold per se did not produce a significant, relative CBFV increase in the contralateral MCA. However, when subjects were expecting a threshold tactile stimulus, there was a significant regional increase in CBFV in the hemisphere contralateral to the attended index finger for approximately 15 s, starting within the first seconds after the cueing. This increase was present even before the tactile stimulus was applied and also in sessions when the stimulus was omitted. We conclude that preparation of the cortex causes a stronger regional cerebral blood flow increase than the processing of the tactile stimulus itself. PMID- 9348130 TI - Intraventricular injection of NGF, but not BDNF, induces rapid motor activation that is inhibited by nicotinic receptor antagonists. AB - The acute and subacute effects of intracerebroventricularly (ICV) administered nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF) on locomotor activity were evaluated in awake adult rats. Immediately after ICV injection through an implanted cannula, locomotor activity was measured by a computerized system using infrared photocells, which allowed us to record locomotion, motility, and rearing simultaneously. A single dose of 5 microg mouse beta-NGF produced significant increases in horizontal ambulatory components of locomotor activity (locomotion and motility), but not vertical movement (rearing) 30-45 min after ICV administration. These increases lasted for at least 3-4 h. Systemic injection of 2.0 mg/kg mecamylamine, a central nicotinic receptor antagonist, inhibited the hyperactivity induced by NGF. Systemic injection of 0.5 mg/kg scopolamine, a muscarinic receptor antagonist, did not interfere with the NGF effects. Thus, while scopolamine induced marked increases in all three measures of behavior in both NGF and cytochrome-c-treated animals, locomotion and motility remained significantly higher in the NGF group. Immunohistochemistry demonstrated that NGF diffused readily from the ventricular space into brain parenchyma on the injected side and could be visualized 1 h after ICV injection. These results suggest that ICV administration of NGF increases locomotor activity by inducing acetylcholine release, and that nicotinic receptors are involved in the hyperactivity induced by NGF. ICV administration of 5 microg recombinant human BDNF had no significant effect on locomotor activity during the 0- to 4-h period after ICV injection. However, it produced significant decreases in locomotion, motility, and rearing 24-26 h later. Hence ICV administration of BDNF has entirely different effects on animal behavior from those evoked by NGF. While NGF elicits increases in ambulatory behavior within hours, BDNF causes delayed decreases in ambulatory behavior. PMID- 9348131 TI - Ascending spinal influences on rubrospinal cells in the cat. AB - Somaesthetic input to rubrospinal cells, bypassing the cerebellum and cerebral cortex, has been demonstrated in the cat. The detailed organization of this somatic afferent system was studied using electrophysiological methods on multiple-lesion, chloralose-anaesthetized preparations. Stimulation of the dorsal column (DC) at upper cervical cord segments induced significant responses in magnocellular red nucleus (RNm) cells in cats without a cerebellum and with ablation of the frontal cortex. As classic descriptions state that primary afferent fibres have ascending and descending branches in the DC, with many collaterals arborizing in the grey matter at the segmental level of the cord, this procedure is equivalent to stimulating the somatic fibres coming from a large portion of the body, leading to the simultaneous activation of most ascending spinal pathways. To show that the pathway responsible for the rubral responses ascends in the ventral spinal cord, and that the synaptic relays are located at the segmental level, the stimulation was applied to the DC, caudally to the sectioned dorsal spinal half. Various tests confirmed that the activation was conducted to rubral cells through antidromically activated primary afferents. Their multiple collaterals relay the messages to cells located caudal to the spinal lesion, with fibres ascending in the ventral cord. Any relay of the somatic rubral responses in the DC's nuclei was excluded. When the DC was sectioned and its rostral end was dissected free and lifted onto two hook electrodes for stimulation, no response was obtained in the rubral cells. This dissection indeed sectioned all DC fibre collaterals entering the grey matter, thus excluding the possibility of segmental relay. Single shocks applied to the ventral quadrant of the cord or in the medial lemniscus (LM) in the medulla oblongata induced monosynaptic excitatory post-synaptic potentials (EPSPs) in most rubrospinal cells. The spinal EPSPs could be collided by stimulation in the LM, thus demonstrating the existence of direct connections from the cord to the RNm. This somaesthetic pathway to the RNm could be involved in on-line correction of movements and in learning new motor strategies. PMID- 9348132 TI - Organization of the nigro-tecto-bulbar pathway to the parvicellular reticular formation: a light- and electron-microscopic study in the rat. AB - We examined a nigro-tecto-bulbar pathway to the parvicellular reticular formation (RFp), where many premotor neurons for orofacial motor nuclei are known to be distributed, by using a combined anterograde and retrograde tracing method. After contralateral injections of biotinylated dextranamine (BDA) into the dorsolateral part of the substantia nigra (SNr) and cholera toxin B subunit (CTb) into the RFp, overlapping distributions of BDA-labeled terminals and CTb-labeled neuronal cell bodies were found in the lateralmost part of the superior colliculus (SC) ipsilateral or contralateral to the site of BDA injection or CTb injection, respectively. After contralateral injections of BDA into the SNr and horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) injection into the RFp, ipsilateral labeled axon terminals with BDA were found to make symmetrical synaptic contacts with the somata and dendrites of contralateral labeled neurons with WGA-HRP in the lateralmost part of the SC. Furthermore, we demonstrated that BDA-labeled axon terminals were immunoreactive for GABA, by using the anterograde tracing method combined with immunohistochemistry for GABA. Thus, GABA-like immunoreactive fibers originating from the dorsolateral part of the SNr make monosynaptic contacts with the tectal neurons sending their axons to the RFp. PMID- 9348133 TI - Synaptic connections from large afferents of wrist flexor and extensor muscles to synergistic motoneurones in man. AB - Short-latency excitatory Ia reflex connections were determined between pairs of human wrist flexor and extensor muscles. Spindle Ia afferents were stimulated by either tendon tap or electrical stimulation. The activity of voluntarily activated single motor units was recorded intramuscularly from pairs of wrist flexor or extensor muscles. Cross-correlation between stimuli and the discharge of the motor units provided a measure of the homonymous or heteronymous excitatory input to a motoneurone. Homonymous motoneurone facilitation was generally stronger than that of the heteronymous motoneurones. The principal wrist flexors, flexor carpi radialis (FCR) and flexor carpi ulnaris (FCU), were tightly connected through a bidirectional short-latency reflex pathway. In contrast, the extensor carpi ulnaris (ECU) and the extensor carpi radialis (ECR) did not have similar connections. ECU motoneurones received no short-latency excitatory Ia input from the ECR. ECR motoneurones did receive excitatory Ia input from ECU Ia afferents; however, its latency was delayed by several milliseconds compared with other heteronymous Ia excitatory effects observed. The wrist and finger extensors were linked through heteronymous Ia excitatory reflexes. The reflex connections observed in humans are largely similar to those observed in the cat, with the exception of heteronymous effects from the ECU to the ECR and from the extensor digitorum communis (EDC) to the ECU, which are present only in humans. The differences in the reflex organization of the wrist flexors versus the extensors probably reflects the importance of grasping. PMID- 9348134 TI - Shoulder and elbow muscle activity in goal-directed arm movements. AB - This research examined the electromyographic (EMG) activity of shoulder and elbow muscles during reaching movements of the upper limb. Subjects performed goal directed arm movements in the horizontal plane. Movements which varied in amplitude, speed, and direction were performed in different sections of the workspace. EMG activity was recorded from the pectoralis major, posterior deltoid, biceps brachii short head, brachioradialis, triceps brachii long head, and triceps brachii lateral head; motion recordings were obtained with an optoelectric system. The analysis focused on the magnitude and timing of opposing muscle groups at the shoulder and elbow joints. For hand movements within any given direction of the workspace direction, kinematic manipulations changed agonist and antagonist EMG magnitude and intermuscle timing in a manner consistent with previous single-joint findings. To produce reaching movements in different directions and areas of the workspace, shoulder and elbow agonist EMG magnitude increased for those hand motions which required higher angular velocities, while the timing between opposing muscle groups at each joint was invariant. PMID- 9348135 TI - Callosal connections of the ferret primary auditory cortex. AB - The callosal connections of ferret auditory cortex were studied by making multiple injections of wheat germ agglutinin-horseradish peroxidase into the middle ectosylvian gyrus or by packing crystals of horseradish peroxidase into the transected corpus callosum. The primary area (AI) had strong callosal connections that arose from somata mainly located in layer III. Other layers contained sparsely distributed cells that projected across the midline. The projecting cells occurred over the whole extent of AI but were not homogeneously distributed in layer III. The axons from these cells terminated mainly in the upper layers of the contralateral cortex, where they converged onto three discrete bands. The three elongated bands lay in a dorsoventral orientation, parallel to the tonotopic axis. They were slightly curved and had a fairly uniform width. The posterior band had a width of about 200 microm, while the anterior and middle bands were more variable and had widths of 300-800 microm. The centre-to-centre distance between the posterior and middle bands was 520+/-60 microm and for the anterior to middle bands was 620+/-210 microm. The retrograde labelling produced by the same injections showed that the cell bodies had a higher density in the terminal bands than in the intervening spaces. The bands of dense callosal connections appear to correspond to the binaural summation columns, which have been clearly demonstrated in the ferret, but direct evidence of this will need to be sought in a future study. The discrete nature of the callosal bands in the ferret appears to make it a suitable species for studying the relationship between callosal terminals and those arising in other areas of the brain and for clarifying the possible existence of separate functional systems within the auditory cortex. PMID- 9348136 TI - Presynaptic inhibition compared with homosynaptic depression as an explanation for soleus H-reflex depression in humans. AB - The H-reflex is depressed for seconds if elicited following a single H-reflex or train of H-reflexes. Presynaptic inhibition from flexor afferents (tibialis anterior) onto soleus Ia afferents elicited by either single or trains of stimuli had no effect on the soleus H-reflex on a time scale of seconds. Postsynaptic inhibition was also excluded by magnetic stimulation tests that showed that the excitability of the motoneuron pool was not changed at latencies within a range of seconds. Homosynaptic depression localized at the presynaptic terminal seems to be the mechanism behind the H-reflex depression in humans. PMID- 9348138 TI - Osteoarticular and muscle infectious lesions in patients with the human immunodeficiency virus. AB - Between 1988 and 1995, 1832 HIV positive patients were evaluated in our institution. We studied the epidemiologic, immunologic and bacteriologic data, laboratory tests, and X-Ray films in those with musculoskeletal infection. We reviewed twenty-one cases of musculoskeletal infection in twenty patients aged 23 35 years (mean 28,6 years, M:F= 15:5). In all of them risk factor for HIV was intravenous drug abuse. The number of CD4 positive lymphocytes ranged from 0,003 to 0,5 10(9)/l. Staphylococcus aureus was the organism responsible of the infection in twelve cases, all active intravenous drug abusers at the time the diagnosis was done. The remaining causative agents were: Mycobacterium tuberculosis (3 cases), Candida albicans (2 cases), Salmonella subgroup 1 (1 case), Neisseria gonorrhoeae (1 case), Pseudomona aeruginosa (1 case) and Streptococcus agalactiae (1 case). Fifteen infections were diagnosed between 1988 and 1991 and 6 between 1992 and 1995. Musculoskeletal infectious lesions in HIV positive patients in our country are related in the majority of cases to intravenous drug abuse. In the last four years due to a National medical health care plan conducted to educate this group of people the number of musculoskeletal infections is decreasing. PMID- 9348137 TI - Effect of dietary selenium and vitamin E on the biomechanical properties of rabbit bones. AB - It is generally agreed that combined deficiency of selenium and vitamin E leads to several abnormalities including Kashin-Beck disease which is an endemic and chronic degenerative osteoarthrosis. The abnormalities can be reversed by the administration of various forms of selenium and vitamin E. The present study was designed to investigate the effects of dietary selenium and vitamin E on bone tissue and on the biomechanical properties of bone. Young rabbits of both sexes were fed with either a selenium- and vitamin E-adequate diet (control group), or a selenium- and vitamin E-deficient diet or a selenium-excess diet. The selenium deficient diet resulted in a significant decrease in plasma selenium level and the selenium-excess diet resulted in a significant increase in the plasma selenium level with respect to the corresponding control values (p < 0.05). The diets did not affect the blood cell counts considerably but erythrocyte glutathione peroxidase activity increased (decreased) relatively when the plasma selenium level increased (decreased) (p < 0.05). The light microscopic investigations of the bone tissues of the two experimental groups indicate that the findings of the present work are compatible with osteomalacia. The biomechanical properties of the bones from the three groups were determined experimentally with bending tests. Both the Se- and vitamin E-deficient diet and the Se-excess diet decreased the biomechanical strength of the bones significantly while the bones belonging to the control group always had the largest modulus of elasticity (p < 0.05). PMID- 9348139 TI - Predictors of survival in 171 patients with systemic sclerosis (scleroderma). AB - Predictors of survival were determined in 171 patients with systemic sclerosis by univariate analysis, and the Cox proportional hazards model using both cross sectional data at entry into the follow-up and time-dependent follow-up data. Clinical and laboratory data were evaluated from 1982 to the end of 1993. The presence of diffuse scleroderma, kidney and cardiac involvements were unfavourable prognostic signs in both the univariate analysis, and the Cox proportional hazards models. The Cox model, using the variables detected at study entry, indicated that pericarditis, and anaemia were bad prognostic signs. Analysis with time dependent data has not been reported in systemic sclerosis. The appearance of pigmentation disturbances, anaemia, and respiratory failure during the follow-up also caused a poor prognosis of the disease by the Cox model. In the stepwise selection models, diffuse scleroderma, internal organ manifestations including renal, and cardiac involvements were predominantly selected as the most unfavourable factors for survival. As to the extent of skin involvement and internal organ manifestations, the general behaviour of the disease seems to be similar throughout the world. The early appearance of pericarditis and pigmentation disturbances at study entry are bad prognostic signs. PMID- 9348141 TI - Cervical spine surgery in ankylosing spondylitis: is the outcome good? AB - OBJECTIVE: To assess retrospectively, the outcome of cervical spine surgery in patients with ankylosing spondylitis (AS). METHODS: A cross-sectional study of 3464 patients with identified AS, 19 patients of whom had cervical spine surgery. A self-administered questionnaire (including the use of 10 cm visual analogue scales, 0 = none, 10 = worst) assessing the complications of the surgery, patients' neck symptoms and post-surgery functional ability was sent to the 19 patients. Available casenotes and radiographs were reviewed. RESULTS: The mean duration of follow-up was 10 years. One patient had two separate cervical spine operations. The types of surgery performed included cervical fusion (n=7), osteotomy (n=7) and laminectomy (n=6). Six patients had minor complications as a result of surgery. The majority of patients (93%) felt that their surgery had been successful. Most patients (81%) had a reduction in neck pain (mean pain score=3.1, SD 2.8) but increased neck stiffness (mean stiffness score=8.0, SD 2.9). Postoperative radiographs of 7 patients showed complete ankylosis of the cervical spine. Generally, few patients reported difficulty with reading/watching television (6%), sleep (19%) or driving (36%). A third of the patients were still in full time employment. CONCLUSIONS: About 1 in 200 patients with AS undergo cervical spine surgery. The surgery is often successful and complications are usually minor. Neck pain is often better after surgery and any remaining neck symptoms do not significantly affect the patient's sleep or functional activities. In this retrospective study, the long term outcome of cervical spine surgery in patients with AS appears to be good. PMID- 9348140 TI - Polymorphism of the LMP2 gene and disease phenotype in ankylosing spondylitis: no association with disease severity. AB - Although a number of reports have now described an association between polymorphism of the LMP2 gene and disease phenotype in HLA-B27 positive individuals with ankylosing spondylitis (AS), some describe associations with acute anterior uveitis, others with juvenile onset disease, and one report provides no association. A recent study describes yet a further association with disease severity in patients with juvenile rheumatoid arthritis. We therefore hypothesized that the discrepant findings in adult disease may be a reflection of an underlying association with disease severity. Our study population consisted of 100 HLA-B27 positive Caucasians with AS of ten or more years duration. Clinical assessment of disease severity was based on a metrology index scoring five measurements, the modified health assessment questionnaire for the spondyloarthropathies, and a disease activity index consisting of a visual analog scale to score the amount of pain, stiffness and fatigue. LMP2 genotypes were assigned following polymerase chain reaction amplification from genomic DNA and restriction enzyme digestion with CfoI. Despite confirmation of a significantly higher prevalence of the LMP2 BB genotype in AAU positive (66.0%) versus AAU negative (45.2%) patients (P < 0.05), we observed no association between LMP2 genotypes and any of the indices of disease severity. Furthermore, although a significant association was noted between the presence of peripheral synovitis and the functional index score (P < 0.05), a history of AAU was not associated with more severe disease. Our data is thus internally consistent in demonstrating no association between LMP2 genotypes and either disease severity or peripheral arthritis, and supports the notion that polymorphism of LMP2 primarily influences the development of AAU and not some other phenotype of AS. PMID- 9348142 TI - The moderate intestinal side effects of auranofin do not require prophylactic therapy with a bulkforming agent. Dutch Ridaura Study Group. AB - Incidences of diarrhoea and loose stools are reported up to 50% in patients starting treatment with auranofin. Moreover, +/-4% of patients discontinue treatment because of severe diarrhoea. We investigated whether a water binding agent would diminish the incidence of loose stools and diarrhoea. Endpoints were the patient's general impression of the quality of stools and a daily assessment of stool's frequency/consistency and adverse events. Secondly, some disease activity parameters were used to evaluate whether the bulkforming agent influences the efficacy of auranofin. In this study 269 patients suffering from Rheumatoid Arthritis (RA) were treated with auranofin 6 mgr daily for a period of six months. Simultaneously the patients were randomly treated with either a bulkforming agent (Volcolon: psyllium fibres) or placebo. Results show a 15% incidence of loose stools and diarrhoea during treatment with auranofin. During the treatment period the patients' general impression of defecation consistency showed a shift to softer types. The changes in defecation consistency was not significantly different between groups (Intention-to-treat analysis: C2=4.01; p=0.13). Also, the percentage of patients experiencing episodes of diarrhoea (reported as an adverse experience) was not different (14% of the patients treated with bulkformer versus 15% with placebo). During the first month 7% (n=5) of placebo treated patients reported short episodes of watery stools versus none in the bulkformer treated group. The percentage of days with loose or watery stools, reported on the diary cards, was consistently lower in bulkformer treated patients. Both groups improved equally with respect to disease activity parameters. Sixty-eight percent of patients continued auranofin treatment after the study period. In conclusion, these data do not support adjuvant therapy with a bulkforming agent on initiation of auranofin therapy. The overall low incidence of loose stools and diarrhoea suggests that a dose increase to 9 mgr daily is an option to enhance the efficacy of auranofin treatment. PMID- 9348143 TI - Destructive pneumococcal septic arthritis in end-stage renal disease. AB - Pneumococcal arthritis generally presents as non-destructive monoarthritis, although some underlying metabolic disorders such as liver failure and diabetes have been suggested to represent a risk factor for severe joint disease. Here we report a case of destructive pneumococcal arthritis of the left hip joint in a patient suffering from chronic renal failure treated with hemodialysis for ten years. Inspite of effective anti-pneumococcal antibiotic treatment, the patient with preexisting renal osteopathy and a mild osteoarthritis continued to suffer from severe and disabling pain of the left hip. This case demonstrates that pneumococcal joint infection in patients with underlying uremic bone disease can lead to quick deterioration of the affected joint. PMID- 9348144 TI - Relapsing polychondritis with Castleman-like lymphadenopathy: a case report. AB - Relapsing Polychondritis (RP) is a systemic disorder characterized by an inflammatory process involving predominantly cartilaginous structures and the cardiovascular system. Lymphadenopathy is a very uncommon finding of RP. We report on a patient affected by RP presenting with lymphadenopathy of Castleman like type quickly responsive to corticosteroids. The bronchial involvement and the evolution of the inflammatory process in a 3-year follow-up has been documented by computed tomography of the chest. PMID- 9348145 TI - Interstitial nephritis, toxic epidermal necrolysis and liver dysfunction associated to fenbufen. PMID- 9348146 TI - Recognition of Sir James Paget (1814-1899) PMID- 9348147 TI - Choices in the 1990s for the management of pediatric cerebral arteriovenous malformations. AB - A retrospective 45-year analysis of the management of 160 children with intracranial arteriovenous malformations at The Hospital for Sick Children, Toronto, reveals substantially improving outcomes which relate to more efficient diagnoses and treatments. 80% of children will declare their malformation by means of spontaneous intracranial hemorrhage. For those children who present with hemorrhage or epilepsy, 80% will require an operation. The overall mortality rate has declined to 12% since 1975 and that for the cerebellar lesions from 67 to 42%. 53% of the patients operated upon will be neurologically normal. Endovascular embolization of a child's AVM is a customized, partial solution for a limited number of children. Stereotactic radiosurgery will be used increasingly to obliterate those small lesions in children which are unassociated with hemorrhage or are the residua of an operation. As many as 10% of children (15/160) with diagnosed AVMs cannot be helped with operative or other interventional therapies. The recognition of the pediatric stroke syndromes, the early triage and diagnosis of a child's cerebral hemorrhage, the operative and anaesthetic technologies and the adjunct therapies - choices of the nineties - have resulted in a 66% decline in the overall mortality from this vascular lesion as well as greater assuredness that for most the lesion can be permanently obliterated. PMID- 9348149 TI - Tethered cord syndrome in low motor level children with myelomeningocele. AB - The clinical presentation of tethered spinal cord and the results of tethered cord release were examined in a group of 30 low motor level (L3 and below) children with a history of myelomeningocele without concomitant CNS complications. Changes in orthopedic and/or neurologic status formed the basis of consideration for tethered cord release. Clinically, these patients presented with a new onset or recently progressing scoliosis, spasticity with or without contractures, decrease in motor function and low back pain at the site of closure. One or more of these findings was present in all cases and led to the suspicion of tethered spinal cord. The diagnosis of tethered cord was confirmed in all cases by MRI or CT myeolography. In order to isolate tethering as the etiology for the patients' clinical deterioration, patients with concomitant CNS complications, e.g. shunt dysfunction or hydromyelia were excluded from the study. Twenty-nine such patients, of an initial 59, who would have otherwise been considered, were excluded on the basis of this criteria of concomitant CNS complications. The results of release 1 year after the procedure were as follows: regarding scoliosis, in 75% of cases the curve either remained stable or decreased by more than 10 degrees, with 25% experiencing curve progression of > 10 degrees. The most recent follow-up in this group revealed that 11.8% experienced a decrease in curvature of >10 degrees; 47.1% remained stable, and 41.2% ultimately progressed 10 degrees. In the group with spasticity, 43.8% improved; 56.3% remained stable, and none worsened. Most (78.6%) of the children who had experienced a decline in motor function improved postoperatively, and all those with back pain experienced complete resolution. In conclusion, tethered cord release in symptomatic low lumbar and sacral level children with myelomeningocele appears to be of benefit, especially with respect to stabilization of scoliosis in selected patients, back pain at the site of closure, and prior decline in motor function. Results in the cases with spasticity were more equivocal. PMID- 9348148 TI - Use of the prone position in the MRI evaluation of spinal cord retethering. AB - In order to determine the impact of magnetic resonance imaging (MRI) in the management of spinal cord retethering, we retrospectively reviewed case and imaging records of 51 patients who underwent MRI examination in supine and prone positions. Group 1 included 8 control patients without cord tethering. They exhibited a normal level of the conus medullaris with normal surrounding subarachnoid space, and consistent anterior migration of the conus within the dural sac on MRI in prone position. Group 2 included 17 patients with tethered cord secondary to occult spinal dysraphism (spinal cord lipoma in 6 patients, thick filum terminale in 4, diastematomyelia in 4, myelomeningocele manque in 2, and dermoid tumour in 1). Supine and prone MRI performed at a median period of time of 6 months after untethering showed resolution of posterior tethering in 5 out of the 7 patients who exhibited pre-operatively dorsal attachment of the spinal cord to the dura. Anterior migration of the conus or of the cord/filum complex in prone position was observed in only 24% of the cases. Group 3 included 26 patients with secondary tethered cord following prior myelomeningocele closure. Their MRI performed at a median interval of time of 11 months following untethering demonstrated resolution of the posterior cord tethering in only 8 out of the 24 patients who exhibited this feature pre-operatively. Anterior migration within the expanded dural sac was never noted in this group. We conclude that spine MRI is of limited value and that prone-positioned MRI is of no additional use in the evaluation of spinal cord retethering. PMID- 9348150 TI - Radiological and clinical criteria for the management of epidural hematomas in children. AB - OBJECTIVE: Modern neuroimaging and intensive care permit precise delineation and specific treatment of head injury. Children sustaining cranial trauma associated with epidural hematoma (EDH) represent a heterogeneous group with a variety of clinical outcomes. Treatment consists of simple observation or surgical evacuation. We attempted to define radiological characteristics of the EDH patients that underwent surgical evacuation. METHODS: We reviewed the records and computed tomography scans of 33 children sustaining cranial trauma associated with EDH treated at the Children's National Medical Center between October 1990 and August 1994. The radiological and clinical characteristics of children treated surgically (n = 13) and nonsurgically (n = 20) were compared. RESULTS: Mass effect, a temporal clot location, thickness, length and volume of the clot, and midline shift (p < 0.05) differed significantly between groups. The most important radiological parameters in determining the therapeutic intervention were thickness, midline shift, mass effect, and EDH location. A thickness of the EDH > 18 mm, a midline shift >4 mm, and moderate or severe mass effect correctly predicted therapy in 29 out of 33 patients. By adding the location as a fourth parameter, therapy was accuratly predicted in 31 of 33 patients. Mechanism of injury, interval from injury to initial computed tomography scan, age, sex, Glasgow coma score on admission, or lengths of hospital and intensive care unit stays were not significantly different between groups. CONCLUSION: Although radiological criteria predict surgical intervention for larger EDH, patients harboring intermediate-size EDH will continue to require careful individualized clinical judgement. PMID- 9348151 TI - Send severely head-injured children to a pediatric trauma center. AB - Between 1985 and 1988, a total of 1,320 head-injured children were seen by the neurosurgical service at the Children's Hospital in Washington, D.C. (CHOW). This included 1,095 minor injuries, 127 moderate injuries, and 98 severe injuries. Children brought from the scene of an accident were considered direct transports, whereas those taken to another hospital prior to transfer to CHOW were indirect. Of the children that suffered severe head injuries, 56 were admitted directly to the hospital and 42 indirectly. The indirect group had a higher percentage of children with lower coma scores and abused children. The trauma score was significantly higher in the direct group (9 vs. 7). The mortality rate for severe injuries referred directly from the scene of the accident was 26.8%, and the mortality rate for children referred indirectly was 50.0%. The higher number of abused children in the indirect group did not account for the increase in mortality rate (p = 0.021) in this group. This is the first study to show that children brought directly from the scene of an accident to a well-established pediatric trauma center have a significantly better chance of survival than children transported first to the nearest available hospital. PMID- 9348152 TI - Urokinase in the treatment of shunt malfunctions caused by thrombus. AB - Thrombus is a frequent cause of shunt malfunction both of the proximal end following intraventricular hemorrhage and of the distal catheter of a vascular shunt. Continued blockages may result in numerous shunt revisions until the blood has been cleared. We have treated 3 children with shunt malfunctions secondary to thrombus with urokinase, a thrombolytic agent. Two children had intraventricular hemorrhage following a shunt revision and were treated with intrashunt urokinase, and 1 with occlusion of an atrial catheter was treated with both intrashunt and systemic urokinase. All were symptomatic at the time of treatment (headaches, vomiting, full fontanel, somnolence) and all had ventriculomegaly demonstrated on computed tomography. Various dosage regimens were used with total intrashunt doses of 20,000, 50,000, and 70,000 IU. All improved clinically, computed tomography scans demonstrated improvement, and all were discharged from the hospital. There were no complications of the urokinase administration. The 2 children with proximal occlusion have not required further shunt revisions at 12 and 27 months following treatment. The infant with atrial end occlusion subsequently underwent two proximal revisions with eventual removal of the atrial catheter because of infection. We conclude the intrashunt urokinase can be of value in the treatment of shunts by blood and blood products. PMID- 9348153 TI - Concerning the article by Duke BJ et al. Pediatr Neurosurg 1996;25:31-35: transdural migration of microfixation plate and screws in the cervical spine. PMID- 9348154 TI - Deep venous thrombosis associated with antiphospholipid antibodies in an adolescent after exeresis of a pilocytic astrocytoma. PMID- 9348155 TI - Concerning the article by Baumann GS et al. Pediatr Neurosurg 1996;24:193-201: gamma knife radiosurgery in children. PMID- 9348156 TI - Endocrine and metabolic manifestations associated with infectious and inflammatory diseases. PMID- 9348157 TI - Microbial factors in disease emergence illustrated by streptococcal toxic shock syndrome. PMID- 9348158 TI - Enterohemorrhagic Escherichia coli in the United States. PMID- 9348159 TI - Cholera: molecular basis for emergence and pathogenesis. PMID- 9348160 TI - Lyme disease (Lyme borreliosis). PMID- 9348161 TI - Tuberculosis in the context of emerging and reemerging diseases. PMID- 9348162 TI - Emergent antibiotic resistance: health risks and economic impact. PMID- 9348163 TI - Influenza: interspecies transmission and emergence of new pandemics. PMID- 9348164 TI - Ebola and hantaviruses. PMID- 9348166 TI - Viral gene expression and pathogenesis in three emerging diseases: HIV and AIDS; HTLV-I and HAM/TSP; and HHV-8 and Kaposi's sarcoma. PMID- 9348165 TI - Impact of dengue virus infection and its control. AB - Dengue virus infection has been counted among emerging and re-emerging diseases because of (1) the increasing number of patients, (2) the expansion of epidemic areas, and (3) the appearance of severe clinical manifestation of dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS), which is often fatal if not properly treated. In the meantime, there are no effective dengue control measures: a dengue vaccine is still under development and vector control does not provide a long-lasting effect. In order to obtain direct evidence for the virulent virus theory concerning the pathogenesis of DHF/DSS, type 2 dengue virus strains isolated from patients with different clinical severities in the same epidemic area in northeast Thailand, during the same season, were comparatively sequenced. The result revealed a DF strain specific amino acid substitution from I to R in the PrM, and a DSS strain specific amino acid substitution from D to G in the NS1 gene regions, which could significantly alter the nature of these proteins. Moreover, DF strain specific nucleotide substitutions in the 3' noncoding region were predicted to alter its secondary structure. These amino acid and nucleotide substitutions in other strains isolated in different epidemic areas during other seasons, together with their biological significance, remain to be confirmed. In order to innovate dengue vector control, field tests were carried out in dengue epidemic areas in Vietnam to examine the efficacy of Olyset Net screen, which is a wide-mesh net made of polyethylene thread impregnated with permethrin. The results show that Olyset Net (1) reduced the number of principal dengue vector species, Aedes aegypti, (2) interrupted the silent transmission of dengue viruses and (3) was highly appreciated by the local people as a convenient and comfortable vector control method. This encouraging evaluation of the Olyset Net screen should be confirmed further by other tests under different settings. PMID- 9348167 TI - Emerging and re-emerging hepatitis viruses. PMID- 9348168 TI - Emerging parasitic infections. PMID- 9348169 TI - Immunoregulation and parasitic infections. PMID- 9348170 TI - Malaria: an emerging and re-emerging global plague. PMID- 9348171 TI - Collaborative research and training programs in infectious diseases: summary of roundtable discussion. PMID- 9348172 TI - The role of scientific societies in addressing the threats of emerging infections: summary of roundtable discussion. PMID- 9348173 TI - When the light turns blue. PMID- 9348174 TI - Identification of the blood-borne somatotroph-differentiating factor during chicken embryonic development. AB - Somatotrophs become a significant population by day 16 of chicken embryonic development. We have previously demonstrated that an earlier induction of GH cell differentiation is possible with the addition of day 16 embryonic serum to cultures of day 12 pituitary cells, an age when somatotrophs are rare. The present study was designed to identify the blood-borne signal(s) responsible for the serum activity, using reverse hemolytic plaque assays to identify individual GH-secreting cells. The activity was found to be a heat-stable, ether-soluble compound(s) that is bound or inhibited by a trypsin-sensitive protein. The extent of GH cell differentiation was greater (P < 0.05; n = 3) in response to the ether phases of heated day 16 (14.1 +/- 0.4% of all cells) and day 12 sera (9.3 +/- 0.4%) than with untreated serum from days 16 and 12 (6.1 +/- 0.4% and 0.82 +/- 0.4%, respectively). Furthermore, ether-extracted day 16 serum was more effective than ether-extracted day 12 serum, which was also different from basal (0.85 +/- 0.4%; P < 0.05). Based on this biochemical profile, the abilities of various steroids to stimulate differentiation were tested. Three steroids were found to stimulate somatotroph differentiation in vitro: 17beta-estradiol, corticosterone, and progesterone. However, the estradiol receptor antagonist, tamoxifen, while abolishing the effect of estradiol, had no effect on the induction of differentiation by day 16 serum. In contrast, RU486, a specific glucocorticoid receptor antagonist in chickens, blocked the stimulatory effects of corticosterone, progesterone, and day 16 serum on somatotroph differentiation. We next tested whether the active compound in day 16 embryonic serum was corticosterone, the predominant glucocorticoid in chickens. Incubation of day 16 serum with corticosterone antiserum, but not control antiserum, suppressed day 16 serum-induced GH cell differentiation. Therefore, we conclude that corticosterone is the blood-borne signal capable of stimulating somatotroph differentiation in vitro. The present findings together with previous reports indicate that somatotroph differentiation during embryonic development may result from an increase in circulating glucocorticoid concentrations. PMID- 9348175 TI - Effects of antagonists of growth hormone-releasing hormone (GHRH) on GH and insulin-like growth factor I levels in transgenic mice overexpressing the human GHRH gene, an animal model of acromegaly. AB - Transgenic mice overexpressing the human GH-releasing hormone (hGHRH) gene, an animal model of acromegaly, were used to investigate the effects of potent GHRH antagonists MZ-4-71 and MZ-5-156 on the excessive GH and insulin-like growth factor I (IGF-I) secretion caused by overproduction of hGHRH. Because metallothionein (MT)-GHRH mice express the hGHRH transgene in various tissues, including the pituitary and hypothalamus, initial experiments focused on the effectiveness of the GHRH antagonists in blocking basal and stimulated GH secretion from pituitary cells in vitro. Both MZ-4-71 and MZ-5-156 suppressed basal release of GH from superfused MT-GHRH pituitary cells, apparently by blocking the action of endogenously produced hGHRH. In addition, these antagonists effectively eliminated the response to stimulatory action of exogenous hGHRH(1-29)NH2 (30 and 100 nM). To ascertain whether MZ-4-71 and MZ-5 156 could antagonize the effect of hGHRH hyperstimulation in vivo, each antagonist was administered to MT-GHRH transgenic mice in a single iv dose of 10 200 microg. Both compounds decreased serum GH levels in transgenic mice by 39-72% at 1 h after injection. The inhibitory effect of 50 microg MZ-5-156 was maintained for 5 h. Twice daily ip administration of 100 microg MZ-5-156 for 3 days suppressed the highly elevated serum GH and IGF-I concentrations in transgenic mice by 56.8% and 39.0%, respectively. This treatment also reduced IGF I messenger RNA levels in the liver by 21.8% but did not affect the level of GH messenger RNA in the pituitary. Our results demonstrate that GHRH antagonists MZ 4-71 and MZ-5-156 can inhibit elevated GH levels caused by overproduction of hGHRH. The suppression of circulating GH concentrations induced by the antagonists seems to be physiologically relevant, because both IGF-I secretion and synthesis also were reduced. Our findings, showing the suppression of GH and IGF-I secretion with GHRH antagonists, suggest that this class of analogs could be used for the diagnosis and therapy of disorders characterized by excessive GHRH secretion. PMID- 9348176 TI - Intrahypothalamic growth hormone feedback: from dwarfism to acromegaly in the rat. AB - Two different dwarf rat models with primary (dw/dw, DW) or secondary (transgenic growth retarded, WF/Tgr) GH deficiency and contrasting hypothalamic GH-releasing hormone (GHRH) and somatostatin (SRIH) expression were implanted sc with GC cells. These form encapsulated rat GH-secreting tumors that maintain high plasma rat GH levels for several weeks. In both strains, GC cell tumors stimulated growth and raised GHBP levels, without affecting pituitary GH content. In DW rats, GC cell implants increased SRIH expression in the periventricular nucleus (PeV), but not in the arcuate nucleus (ARC), whereas their high GHRH expression in ARC was decreased by GC cells. In contrast, GC cell implants in WF/Tgr rats had little effect on the already high SRIH expression in PeV or low GHRH expression in ARC, although they reduced SRIH expression in ARC. GC cell implants also reduced GH receptor expression in both ARC and PeV in the WF/Tgr dwarves. Thus, chronic GH overexposure stimulates rapid growth in both dwarf strains, but has differential hypothalamic effects in these models. This experimental approach now makes it possible to study the effects of pathophysiological concentrations of GH ranging from dwarfism to acromegaly in the same animal model. PMID- 9348177 TI - Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat. AB - Synthetic GH secretagogues (GHSs) act via a receptor (GHS-R) distinct from that for GH-releasing hormone (GHRH). We have studied the hypothalamic expression and regulation of this receptor by in situ hybridization using a homologous riboprobe for rat GHS-R. GHS-R mRNA is prominently expressed in arcuate (ARC) and ventromedial nuclei (VMN) and in hippocampus, but not in the periventricular nucleus. Little or no specific hybridization could be observed in the pituitary under the conditions that gave strong signals in the hypothalamus. No sex difference in GHS-R expression was found in ARC or hippocampus, though expression in VMN was lower in males than in females. Compared with GHRH and neuropeptide Y (NPY), GHS-R was expressed in a distinct region of ventral ARC, and in regions of VMN not expressing GHRH or NPY. GHS-R expression was highly sensitive to GH, being markedly increased in GH-deficient dw/dw dwarf rats, and decreased in dw/dw rats treated with bovine GH (200 microg/day) for 6 days. Similar changes were observed in GHRH expression, whereas NPY expression was reduced in dw/dw rats and increased by bGH treatment. Continuous sc infusion of GHRP-6 in normal female rats did not alter ARC or VMN GHS-R expression. Our data implicate ARC and VMN cells as major hypothalamic targets for direct GHS action. The sensitivity of ARC GHS-R expression to modulation by GH suggests that GHS-Rs may be involved in feedback regulation of GH. PMID- 9348178 TI - Fas antigen-mediated apoptosis of ovarian surface epithelial cells. AB - The Fas antigen is a cell surface receptor that, when engaged by Fas ligand or specific agonistic antibodies, triggers apoptosis. The effect of an agonistic monoclonal antibody to mouse Fas antigen (Fas mAb, clone J02) on the viability of cells from dispersed mouse corpora lutea (CL cultures) was tested. Cultures were prepared by enzymatic digestion of CL from day 4-7 pseudopregnant mice. Cultures were pretreated with 0, 1, 10, 100, or 1000 U/ml murine interferon-gamma (IFN) at 72 h of culture. IFN has been shown to increase Fas antigen expression in a number of cell types. At 96 h (time zero), cultures were treated with Fas mAb or IgG. By 4 h after Fas mAb treatment, discrete homogeneous patches of cells within the cultures showed characteristic signs of apoptosis, including blebbing of cell membranes, detachment, and disappearance from the culture. CL cultures contain luteal, stromal, and endothelial cells; fibroblasts; and surface epithelial cells (OSE). Cells dying in response to Fas mAb were identified as OSE. Affected cells had the cobblestone appearance and distinct nuclei typical of epithelial cells. Unlike luteal cells, OSE did not stain with the lipophilic dye, Nile red. The cells did not stain with acetylated low density lipoprotein conjugated to the fluorescent marker octadecyl indocarbocyanine, a marker for endothelial cells and monocytes. Cells in patches stained positively for cytokeratin, a marker for epithelial cells. Fas-mediated cytotoxicity was quantified by counting the number of cells present in discrete patches of OSE 0 and 8 h after Fas mAb treatment. Fas mAb treatment had no effect in cultures pretreated with 0 or 1 U/ml IFN, but induced significant death of OSE in cultures pretreated with 10, 100, and 1000 U/ml IFN (37 +/- 11%, 54 +/- 18%, and 60 +/- 11%, respectively). There was no apparent effect of Fas mAb on other cell types within the CL cultures. To confirm that cells dying in response to Fas mAb were OSE, experiments were also performed on enriched cultures of OSE prepared by enzymatic digestion of the outer surface of the ovary. In enriched OSE cultures pretreated with 200 U/ml IFN, there was 44% killing in response to Fas mAb, whereas in cells not pretreated with IFN, there was no effect. In situ fluorescent end labeling of DNA in CL cultures indicated that treatment with IFN and Fas mAb induced DNA fragmentation in OSE typical of apoptosis. Immunocytochemistry of CL cultures indicated that Fas antigen was expressed in OSE pretreated with IFN. Quantitative reverse transcriptase-PCR showed that IFN pretreatment increased Fas antigen messenger RNA levels 2.3-fold in enriched cultures of OSE. In summary, OSE in CL cultures and enriched cultures of OSE undergo apoptosis in response to Fas mAb when pretreated with IFN. In vivo, OSE undergo programmed cell death before ovulation and rapidly proliferate to repair the surface of the ovulatory follicle after ovulation. Most ovarian cancers are derived from the OSE. The results have implications for both normal ovarian function and oncogenesis in the ovary. PMID- 9348179 TI - 17Beta-estradiol antagonizes effects of 1alpha,25-dihydroxyvitamin D3 on interleukin-6 production and osteoclast-like cell formation in mouse bone marrow primary cultures. AB - In mouse bone marrow primary cultures, the formation of osteoclast-like, i.e. tartrate-resistant acid phosphatase (TRAP)- and calcitonin receptor-positive multinucleated cells (MNC), when induced by 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3), can be suppressed by 17beta-estradiol (17beta-E2), whereas 17alpha-E2 is without any effect. 17beta-E2, above 10(-11) M, significantly reduced 1alpha,25(OH)2D3-mediated TRAP+ MNC formation in cultured bone marrow cells from both female and male mice. The estrogen at 10(-8) M suppressed the peak response to the vitamin D sterol by 50%. 17beta-E2 significantly suppressed basal and 1alpha,25(OH)2D3-stimulated cellular production of interleukin (IL)-6. IL-6 alone, although bone marrow cells in hormone-free culture produced appreciable amounts of the cytokine, did not induce any TRAP+ MNC. Therefore, the changes in IL-6 production induced by the hormones could not be the sole determinant for the extent of TRAP+ MNC formation. However, the stimulatory effect of 1alpha,25(OH)2D3 on osteoclastogenesis nevertheless can be significantly reduced by a neutralizing monoclonal anti-IL-6 antibody. In the presence of 10(-8) M 17beta-E2, the anti-IL-6 monoclonal antibody does not achieve any further suppression of 1alpha,25(OH)2D3-related osteoclast-like cell formation. Our data suggest that induction of osteoclastogenesis by 1alpha,25(OH)2D3 is partially dependent on IL-6 signaling and can be modulated by 17beta-E2 through interference with IL-6 receptor activation, in addition to inhibition of IL-6 production by marrow stromal cells. PMID- 9348180 TI - Paradoxical action of activin A on folliculogenesis in immature and adult mice. AB - Activin A is a gonadal protein originally isolated from follicular fluid and is recognized as a local regulator of granulosa cell differentiation. Whether activin A promotes folliculogenesis, however, still remains unclarified. The present study was designed to elucidate the effect of activin A on follicular growth in in vitro follicle culture systems. Preantral follicles, 100-120 microm in diameter, were mechanically isolated from BDF1 hybrid immature mice (11 days old) and adult mice (8 weeks old), then cultured for 4 days in a serum-free medium supplemented with activin A (100 ng/ml), FSH (100 mIU/ml), and a combination of both. Follicular diameter was measured daily, and the amount of estradiol and inhibin released at day 4 was determined by RIA. Preantral follicles collected from immature mice showed a significant increase in diameter when cultured with activin A or both activin A and FSH. FSH alone showed no significant effect on the diameter of follicles from immature mice. In contrast, the diameter of preantral follicles from adult mice significantly increased in response to FSH. Activin A did not stimulate growth of follicles from adult mice, and more interestingly, blocked the effect of FSH. The inhibitory action of activin A was in part restored by co-culture with follistatin (100 ng/ml). These results indicate that activin A is folliculogenetic in the prepubertal mouse, but not in adults. PMID- 9348182 TI - Biphasic regulation of the preproendothelin-1 gene by c-myc. AB - Endothelin-1 (ET-1), a potent vasoconstrictive/mitogenic peptide originally isolated from vascular endothelium, stimulates the expression of immediate early response genes such as c-myc. The c-myc protooncogene participates in regulating the cascade of events that follow mitogenic stimulation of quiescent cells. Using a panel of isogenic fibroblast cell lines with differential c-myc expression levels (obtained by disrupting one c-myc gene copy with targeted homologous recombination and subsequently stably transfecting the heterozygous cells with an exogenous c-myc transgene), we demonstrate that c-Myc protein regulates ET-1 gene transcription in a biphasic fashion: as an activator at low concentrations and as a repressor at high concentrations. Using rat endothelial cells treated with antisense c-myc oligodeoxynucleotides, we also show that c-myc regulates ET-1 synthesis and secretion in a biphasic manner. The present report, therefore, demonstrates the existence of a signal transduction pathway that regulates the synthesis and secretion of ET-1 via the immediate early transcription factor, c Myc. PMID- 9348181 TI - Retrovirus-mediated herpes simplex virus thymidine kinase gene transduction renders human thyroid carcinoma cell lines sensitive to ganciclovir and radiation in vitro and in vivo. AB - In an attempt to develop gene therapy for thyroid carcinomas, the present studies were undertaken to evaluate in vitro and in vivo therapeutic efficacy and toxicity of herpes simplex virus thymidine kinase (HSV-tk) gene and ganciclovir (GCV) treatment, a widely used prodrug/suicide gene therapy, in human thyroid carcinoma cell lines, FRO and WRO cells, using a means of retrovirus-mediated gene transduction. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the HSV-tk gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) shifted from 250 to 0.5 mg/liter in FRO cells, and from 3,000 to 0.09 mg/liter in WRO cells with therapeutic indexes of 500 and 33,000, respectively. Treatment with 30 mg/liter GCV for 4 days led to complete cell death in HSV-tk tumor cells. Nontransduced cells mixed with transduced cells were also effectively killed by GCV (bystander effect). Low concentrations of GCV, which alone showed little cytotoxicity, enhanced radiation-induced cytotoxicity (radiosensitization). In vivo sc FRO-tk tumor models in nude mice also showed dose- and time-dependent tumor regression. The IC50 was less than 2 mg/kg, and treatment with 100 mg/kg GCV for 2 weeks completely eradicated all tumors. The bystander effect and radiosensitization were also obtained in vivo. These results suggest that the HSV tk/GCV approach to human thyroid carcinoma cells appears to be very efficacious, with a wide therapeutic range, and exerts a bystander effect and radiosensitization both in vitro and in vivo. Thus, HSV-tk/GCV system, alone or in combination with radiotherapy, may be a promising suicide gene therapy for thyroid carcinomas. PMID- 9348183 TI - Localization and differential expression of adenylyl cyclase messenger ribonucleic acids in rat adrenal gland determined by in situ hybridization. AB - The expression of adenylyl cyclases (ACs) in the adult rat adrenal gland was examined. In situ hybridization revealed specific patterns of AC messenger RNA (mRNA) distribution. AC1 was limited exclusively to the adrenal medulla. AC5 and AC6 were mainly expressed in the adrenal medulla, with a weak expression in the zona glomerulosa. AC9 was found in all the three regions of the adrenal cortex but not in the adrenal medulla. All these ACs were detected on postnatal day 1 (PN1), and their pattern of expression was unchanged on PN7, PN21, and PN90 (adult). We analyzed the response of these ACs to various physiological conditions known to affect the synthesis of aldosterone and corticosterone in the adrenal cortex. Our study demonstrates a specific increase of AC6 but not AC5 mRNA in the zona glomerulosa of rats given a low sodium diet. AC9 mRNA was increased in all the three cortical zones of rats treated with ACTH. We suggest that AC6 and AC9 play important roles in different pathways associated with the regulation of aldosterone and corticosteroid production. PMID- 9348184 TI - Prostaglandin E2 receptor subtype EP2 gene expression in the mouse uterus coincides with differentiation of the luminal epithelium for implantation. AB - Among the PGs, PGE2 is considered especially important for implantation and decidualization. Four major PGE2 receptor subtypes, EP1, EP2, EP3, and EP4, mediate various PGE2 effects via their coupling to distinct signaling pathways. Previously, we have shown that the EP1, EP3, and EP4 genes are expressed in the periimplantation mouse uterus in a spatio-temporal manner, suggesting compartmentalized actions of PGE2 during this period. In this study, we examined the expression of the EP2 gene in the mouse uterus during the periimplantation period (days 1-8) and during experimentally induced progesterone (P4)-maintained delayed implantation and its resumption by 17beta-estradiol (E2). We also examined its regulation in the uterus by ovarian steroid hormones. Our results establish that EP2 messenger RNA (mRNA) is expressed exclusively in the luminal epithelium primarily on day 4 (the day of implantation) and day 5 (early implantation) of pregnancy. In (P4)-maintained delayed implanting mice, EP2 mRNA was present in the luminal epithelium, and the expression was further enhanced regardless of the location of the blastocysts after reinitiation of implantation. This observation suggests little or no embryonic influence in regulating EP2 expression and, instead, shows its regulation by P4 and E2. Indeed, treatment with E2 and/or P4 exhibited unique regulation of this gene. The treatment of adult ovariectomized mice with E2 down-regulated the basal levels of EP2 mRNA, whereas that with P4 up-regulated its levels in the luminal epithelium. The up regulation of EP2 mRNA levels by P4 was further augmented by superimposition of the E2 treatment, suggesting a synergistic interaction between E2 and P4 in regulating this gene in the uterus. Collectively, the results suggest that EP2 could be a potential mediator of PGE2 actions in regulating luminal epithelial differentiation and serve as a marker for uterine receptivity for implantation. PMID- 9348185 TI - The effects of programmed administration of human parathyroid hormone fragment (1 34) on bone histomorphometry and serum chemistry in rats. AB - PTH treatment can result in dramatic increases in cancellous bone volume in normal and osteopenic rats. However, this potentially beneficial response is only observed after pulsatile treatment; continuous infusion of PTH leads to hypercalcemia and bone abnormalities. The purpose of these studies was to determine the optimal duration of the PTH pulses. A preliminary study revealed that human PTH-(1-34) (hPTH) is cleared from circulation within 6 h after sc administration of an anabolic dose of the hormone (80 microg/kg). To establish the effects of gradually extending the duration of exposure to hPTH without increasing the daily dose, we programmed implanted Alzet osmotic pumps to deliver the 80 microg/kg x day dose of the hormone during pulses of 1, 2, and 6 h/day, or 40 microg/kg x day continuously. Discontinuous infusion was accomplished by alternate spacing of external tubing with hPTH solution and sesame oil. After 6 days of treatment, we evaluated serum chemistry and bone histomorphometry. As negative and positive controls, groups of rats received pumps that delivered vehicle only and 80 microg/kg x day hPTH by daily sc injection, respectively. Dynamic and static bone histomorphometry revealed that the daily sc injection and 1 h/day infusion dramatically increased osteoblast number and bone formation in the proximal tibial metaphysis, whereas longer infusion resulted in systemic side effects, including up to a 10% loss in body weight, hypercalcemia, and histological changes in the proximal tibia resembling abnormalities observed in patients with chronic primary hyperparathyroidism, including peritrabecular marrow fibrosis and focal bone resorption. Infusion for as little as 2 h/day resulted in minor weight loss and changes in bone histology that were intermediate between sc and continuous administration. The results demonstrate that the therapeutic interval for hPTH exposure is brief, but that programmed administration of implanted hormone is a feasible alternative to daily injection as a route for administration of the hormone. PMID- 9348186 TI - Tissue distribution and quantitative analysis of estrogen receptor-alpha (ERalpha) and estrogen receptor-beta (ERbeta) messenger ribonucleic acid in the wild-type and ERalpha-knockout mouse. AB - Until recently, only a single type of estrogen receptor (ER) was thought to exist and mediate the genomic effects of the hormone 17beta-estradiol in mammalian tissues. However, the cloning of a gene encoding a second type of ER, termed ERbeta, from the mouse, rat, and human has prompted a reevaluation of the estrogen signaling system. Based on in vitro studies, the ERbeta protein binds estradiol with an affinity similar to that of the classical ER (now referred to as ERalpha) and is able to mediate the effects of estradiol in transfected mammalian cell lines. Essential to further investigations of the possible physiological roles of ERbeta, and its possible interactions with ERalpha, are data on the tissue distribution of the two ER types. Herein, we have described the optimization and use of an RNase protection assay able to detect and distinguish messenger RNA (mRNA) transcripts from both the ERalpha and ERbeta genes in the mouse. Because this assay is directly quantitative, a comparison of the levels of expression within various tissues was possible. In addition, the effect of disruption of the ERalpha gene on the expression of the ERbeta gene was also investigated using the ERalpha-knockout (ERKO) mouse. Transcripts encoding ERalpha were detected in all the wild-type tissues assayed from both sexes. In the female reproductive tract, the highest expression of ERbeta mRNA was observed in the ovary and showed great variation among individual animals; detectable levels were observed in the uterus and oviduct, whereas mammary tissue was negative. In the male reproductive tract, significant expression of ERbeta was seen in the prostate and epididymis, whereas the testes were negative. In other tissues of both sexes, the hypothalamus and lung were clearly positive for both ERalpha and ERbeta mRNA. The ERKO mice demonstrated slightly reduced levels of ERbeta mRNA in the ovary, prostate, and epididymis. These data, in combination with the several described phenotypes in both sexes of the ERKO mouse, suggest that the biological functions of the ERbeta protein may be dependent on the presence of ERalpha in certain cell types and tissues. Further characterization of the physiological phenotypes in the ERKO mice may elucidate possible ERbeta specific actions. PMID- 9348187 TI - Maternal deprivation and stress induce immediate early genes in the infant rat brain. AB - The hypothalamic-pituitary-adrenal (HPA) axis is normally quiescent during the stress-hyporesponsive period (SHRP) from day 4-14 in infant rats. However, maternal deprivation (DEP) can disinhibit the HPA axis, thus enabling neonatal rats to respond to mild stressors. In an effort to understand how DEP may alter HPA axis sensitivity, we used in situ hybridization to measure changes in the expression of stress-responsive genes in the brains of neonatal rats. Despite the minimal HPA axis response in nondeprived rats during the SHRP (postnatal day 12), the mild stress of a saline injection significantly increased messenger RNA levels of two immediate-early genes (IEGs), c-fos and NGFI-B, in the hypothalamic paraventricular nucleus (PVN) and in the cerebral cortex. Following 24 h of DEP, the induction of IEGs in response to stress was greatly potentiated in the PVN of P12 neonates. In contrast, DEP attenuated the effects of stress on IEG induction in rats that had matured beyond the SHRP (P20). Surprisingly, DEP decreased basal levels of CRH messenger RNA in the PVN at P12 and P20. Thus the SHRP most accurately refers to HPA axis insensitivity to stress because the brain itself readily responds to stress as evidenced by the induction of IEGs. PMID- 9348188 TI - Short-term treatment of rats with high dose 1,25-dihydroxyvitamin D3 stimulates bone formation and increases the number of osteoblast precursor cells in bone marrow. AB - Using an experimental rat model, this study was undertaken to assess the effects of a short-term application of high dose 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on calcium homeostasis, cancellous bone formation, and numbers of osteoblast precursors in ex vivo bone marrow cultures. For Exp 1 and 2, 6-month-old female rats were sc injected with either 0.2 microg 1,25-(OH)2D3/kg x day or vehicle on days 1, 2, and 3 of the studies. Serum calcium and urinary excretion of calcium were monitored for 12 days in Exp 1. In Exp 2, the rats were ip labeled with five different fluorochromes on days 0, 5, 10, 15, and 20, respectively. Half of the rats in each group were killed on day 7, the rest of the rats were killed on day 24, and the first lumbar vertebrae were processed for histomorphometry. In Exp 3, 0.2 microg 1,25-(OH)2D3/kg BW or vehicle was sc administered to 6-month-old male rats on days 1, 2, and 3, and the number of colony-forming units with the ability to express alkaline phosphatase, to calcify, and/or to synthesize collagen were enumerated sequentially on days 4, 6, 8, 10, 12, and 14 in bone marrow cultures. Short-term 1,25-(OH)2D3 treatment resulted in increased values for serum and urinary calcium during the treatment phase in Exp 1, depressed osteoclast numbers and strongly elevated osteoblast perimeter by day 7, and stimulated mineral apposition rate and bone formation rate in the interval between days 5-15 of Exp 2. Moreover, 1,25-(OH)2D3 administration to rats significantly enhanced the number of mesenchymal precursor cells in bone marrow with the ability to differentiate into an osteoblastic phenotype in ex vivo bone marrow cultures on day 4 of Exp 3. These studies provide evidence that short-term 1,25-(OH)2D3 treatment creates new bone remodeling units and augments osteoblast recruitment and osteoblast team performance in rat cancellous bone. PMID- 9348189 TI - Signaling pathways for guanylin and uroguanylin in the digestive, renal, central nervous, reproductive, and lymphoid systems. AB - Guanylin and uroguanylin are peptides that stimulate membrane guanylate cyclases (GC) and regulate intestinal and renal function via cGMP. Complementary DNAs were isolated encoding opossum preproguanylin and a 279-amino acid portion of a receptor-guanylate cyclase expressed in opossum kidney (OK) cells (GC-OK). The tissue expression of messenger RNA transcripts for these signaling molecules were then compared. Northern and/or reverse transcription-PCR assays revealed that guanylin, uroguanylin, and GC-OK messenger RNAs are expressed in tissues within the digestive, renal, central nervous, reproductive, and lymphoid organ systems. Receptor autoradiography localized the receptors for uroguanylin and guanylin to renal proximal tubules and seminiferous tubules of testis. Synthetic guanylin and uroguanylin peptides activated the receptor-GCs in opossum kidney cortex and in cultured OK cells eliciting increased intracellular cGMP. Expression of agonist and receptor-GC signaling molecules provides a pathway for paracrine and/or autocrine regulation of cellular functions via cGMP in the digestive, renal, central nervous, reproductive, and lymphoid/immune organ systems. Uroguanylin also links the intestine and kidney in a potential endocrine axis that activates tubular receptor-GCs and influences renal function. PMID- 9348190 TI - An estrogen receptor binding site within the human galanin gene. AB - Regulation of galanin gene expression in the anterior pituitary (AP) is positively influenced by estrogen in rodents and undetermined in humans. The objective of this study was to investigate the mechanism behind estrogen induction of galanin by identifying any putative estrogen receptor (ER) binding sequences within the human galanin promoter that may function as estrogen response elements (ERE). Two regions, gERE1 and gERE2, were identified in the galanin 5'-flanking sequence with similarity to the full 13-base ERE consensus previously defined in the vitellogenin gene (vERE). Both sequences were tested in mobility shift assays for the ability to bind nuclear proteins isolated from rat AP tissue or MtTW-10 pituitary tumors. Only the distal sequence at -527 (gERE1) yielded an ERE-specific DNA/protein complex distinguished by mobility and cross competition with vERE. The gel mobility pattern of the DNA/protein complex was comparable between the pituitary tissue and tumor extracts. However, DNA/protein affinity estimations demonstrated a greater affinity of pituitary proteins for gERE1 over the vERE sequence. Evidence that the human ER (hER) does recognize the gERE1 sequence in the human galanin gene was provided by electrophoretic mobility shift assays (EMSAs) with Sf9 extracts enriched in recombinant hER. In addition, antibodies specific for the hER recognized the gERE1/protein complex in supershift experiments. Enhancer activity by gERE1 was detected in transient transfections of the rat GH3 pituitary cell line, resulting in a 4-fold induction of expression driven by the heterologous thymidine kinase promoter in the presence of estrogen. Evidence for ER regulation of the gERE1 enhancer was demonstrated by: 1) inhibition of enhancement using the specific ER antagonist ICI 164,384; and 2) enhancement in HeLa cells that was dependent upon coexpression with hER. Enhancement by gERE1 was half the magnitude as that from the vERE element and may reflect a difference in affinity or composition of the ER complex between the two sequences. These data demonstrate the presence of a functional ERE sequence within the human galanin gene that could potentially function as a regulatory element for estrogen action in the AP. PMID- 9348191 TI - Transforming growth factor-beta1 inhibits membrane association of protein kinase C alpha in a human prostate cancer cell line, PC3. AB - The postreceptor signaling pathway(s) that mediates the effects of transforming growth factor-beta1 (TGF-beta1) is incompletely understood. The present study investigated the involvement of protein kinase C (PKC) in the growth-inhibitory action of TGF-beta1 in PC3, a human prostate cancer cell line. PKC alpha, the only conventional PKC isoform detected in PC3 cells, appeared to be constitutively active based on its presence in both Triton-soluble membrane fraction and cytosol. However, levels of membrane-associated PKC alpha were decreased by a growth-inhibitory dose of TGF-beta1. The response to TGF-beta1 was rapid (within 5 min), time dependent, isoform specific, and occurred without apparent changes in levels of total PKC alpha protein. TGF-beta1 also decreased the levels of membrane-associated PKC activity coincident with its inhibitory effect on PKC alpha's membrane association. Inhibition of PKC activity appeared to be associated with growth inhibition in PC3 cells, because chelerythrine (a specific PKC inhibitor) likewise decreased cell proliferation. Taken together, our data suggest that inhibition of PKC activity, at least in part due to inactivation of PKC alpha, is an early event associated with TGF-beta1 postreceptor signaling that might mediate suppression of cell proliferation. PMID- 9348192 TI - High fat diets elevate adipose tissue-derived tumor necrosis factor-alpha activity. AB - Adipose tissue-derived tumor necrosis factor-alpha (AT-TNF) has been associated with genetic models of insulin resistance and obesity. It is presently unknown if secreted AT-TNF protein is bioactive or whether it can be increased by environmentally induced obesity. In this study, male Wistar rats were fed either a low fat (LF; 12% of energy from corn oil) or a high fat (HF; 45% of energy from corn oil) diet for 5 weeks. From previous data, it is known that after 3 weeks, HF fed animals are obese and insulin resistant compared with the LF group. Hence, animals were killed at 1 week of HF feeding, during the acute response to the diet, and at 5 weeks, when differences in body fat are manifest. Weight gain was significantly increased by diet (P = 0.03) and time (P < 0.0001). AT-TNF bioactivity was measured on secreted protein collected from medium of minced, incubated epididymal (EPI), mesenteric (MES), and retroperitoneal (RETRO) fat pads. AT-TNF bioactivity was significantly increased by diet (P = 0.003) in the RETRO pad and tended to increase (P = 0.07) in EPI. AT-TNF activity was unaffected by diet or time in the MES pad. In the RETRO pad, TNF activity correlated negatively with RETRO fat cell number (r = -0.46, P = 0.002). Secreted AT-TNF protein did not correlate with AT-TNF activity but instead decreased in RETRO with time but not diet. In EPI, secreted AT-TNF protein decreased with the HF diet. Thus, these data suggest that high fat diets and obesity can influence AT-TNF bioactivity and secretion but in an apparent fat pad-specific manner. PMID- 9348193 TI - Transforming growth factor-beta1 regulation of prostaglandin G/H synthase-2 expression in osteoblastic MC3T3-E1 cells. AB - Transforming growth factor-beta (TGFbeta) plays an important role in bone development and remodeling. TGFbeta stimulates PGE2 production, enhances interleukin-1-stimulated PGE2 production, and can stimulate PG-mediated bone resorption. We found that TGFbeta induced prostaglandin G/H synthase (PGHS-2) messenger RNA (mRNA) and PGE2 production in neonatal mouse calvarial cultures and in primary cells derived from these calvariae. We used MC3T3-E1 cells, an immortalized osteoblastic cell line derived from mouse calvariae, to examine the mechanism of PGHS-2 induction. PGHS-2 mRNA was rapidly induced by TGFbeta (10 ng/ml) in MC3T3-E1 cells; mRNA levels peaked at 4-8 h and were still elevated at 24 h. Induction of PGHS-2 protein and PGE2 production correlated with PGHS-2 mRNA levels. In contrast, TGFbeta had much less effect on PGHS-1 mRNA levels. Unlike the response to other agonists, PGHS-2 mRNA induction by TGFbeta was not enhanced by cycloheximide pretreatment, suggesting a requirement for new protein synthesis. To study transcriptional regulation, cells were stably transfected with a PGHS-2 promoter-luciferase reporter construct containing 371 bp of the 5' flanking region and 70 bp of untranslated DNA from the PGHS-2 gene. TGFbeta stimulated luciferase activity paralleled PGHS-2 mRNA induction. Stimulation of luciferase activity and PGHS-2 mRNA levels by other agonists, including interleukin-1, TGF alpha, forskolin, and phorbol 13-myristate 12-acetate, were enhanced by TGFbeta. A 90% drop in luciferase activity occurred with deletion of the region from -371 to -213 bp of the PGHS-2 promoter. The PG response to TGFbeta in MC3T3-E1 cells appears to be mediated primarily by transcriptional regulation of PGHS-2 expression through one or more cis-acting elements located between -371 and -213 bp in the 5'-flanking region of the PGHS-2 gene. PMID- 9348194 TI - Insulin-like growth factors stimulate expression of hepatocyte growth factor but not transforming growth factor beta1 in cultured hepatic stellate cells. AB - Hepatic stellate cells (HSC) are located adjacent to hepatocytes and produce hepatocyte growth factor (HGF) in the normal liver, whereas transformed HSC in fibrotic livers produce transforming growth factor beta1 (TGFbeta1), an inhibitor ofhepatocyte proliferation. In addition to the endocrine actions of hepatic insulin-like growth factor-I (IGF-I), it also stimulates the proliferation of HSC. In this study we found that addition of IGF-1 (20-500 ng/ml) for 48 h to 2- to 7-day-old primary cultures of rat HSC resulted in a time- and dose-dependent increase by 50-190% of the concentrations of immunoreactive HGF in the medium. The levels of HGF as well as DNA synthesis measured as thymidine incorporation were also enhanced by IGF-II and des(1-3)IGF-I, which has reduced binding to IGF binding proteins. There was no consistent effect of the IGFs on the levels of immunoreactive TGFbeta1 or on the total DNA content of the cultures. There was no effect of human GH on medium levels of HGF or TGFbeta1, thymidine incorporation, or total DNA content. IGF-I increased the abundance of HGF messenger RNA, as measured by the RNase protection/solution hybridization technique, whereas there was no effect on TGFbeta1 or glyceraldehyde phosphate dehydrogenase messenger RNA. The results suggest that IGFs stimulate the production of HGF but not TGFbeta1 by HSC in vitro. PMID- 9348195 TI - Differential effects of 1,25-dihydroxyvitamin D3 and tetradecanoylphorbol acetate on cell cycle and apoptosis of MCF-7 cells and a vitamin D3-resistant variant. AB - 1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3), the active form of vitamin D3, and tetradecanoylphorbol acetate (TPA) are potent negative growth regulators of breast cancer cells. In this study, we compared the mechanism of action of these two compounds in MCF-7 cells and a vitamin D3-resistant variant (MCF-7D3Res). In parental MCF-7 cells, 1,25-(OH)2D3 induced morphological and biochemical markers of apoptosis (chromatin and nuclear matrix condensation and DNA fragmentation), whereas TPA induced growth arrest without apoptosis. Both 1,25-(OH)2D3 and TPA independently up-regulated the vitamin D receptor, p21, and the hypophosphorylated form of retinoblastoma (Rb) protein. The growth regulatory effects of 1,25-(OH)2D3 and TPA did not correlate with induction of p53 protein expression. When both compounds were added simultaneously, synergistic effects on MCF-7 cell number were observed, and cell cycle regulatory proteins were down regulated. The MCF-7D3Res cells, which are not sensitive to 1,25-(OH)2D3, were growth inhibited by TPA, and TPA partially sensitized MCF-7D3Res cells to the growth inhibitory effects of 1,25-(OH)2D3. In MCF-7D3Res cells, 1,25-(OH)2D3 treatment had minimal effects on p21 or Rb protein expression, whereas TPA down regulated Rb protein and transiently up-regulated p21. These studies indicate dissociation between the pathways triggered by 1,25-(OH)2D3 and TPA, which mediate growth regulation in MCF-7 cells. Because both compounds induce growth arrest, but only 1,25-(OH)2D3 mediates apoptosis, we conclude that cell cycle arrest is not sufficient to trigger cell death of MCF-7 cells, and that 1,25 (OH)2D3 generates distinct signals which lead to induction of apoptosis in breast cancer cells. PMID- 9348196 TI - Simultaneous measurement of gonadotropin-releasing hormone in the third ventricular cerebrospinal fluid and hypophyseal portal blood of the ewe. AB - GnRH is present in the hypophyseal portal blood and cerebrospinal fluid (CSF) of several species investigated, including sheep, but the precise relationship between these two compartments of GnRH is unknown. In the present study, ovariectomized steroid-treated ewes were surgically prepared for the simultaneous collection of portal blood and third ventricular CSF. Ten-minute samples were collected for pulse analysis after progesterone removal and hourly for comparisons during the estradiol-induced LH surge. The time of onset of the portal (15.3 +/- 0.5 h after estradiol) and CSF (15.9 +/- 0.2 h) GnRH surges was similar and occurred coincidentally with the LH surge (15.6 +/- 0.4 h). The period of the surge during which GnRH concentrations exceeded half-maximal levels (portal, 7.3 +/- 1.5 h; CSF, 7.3 +/- 0.3 h) was the same and outlasted the corresponding LH surge period (3.3 +/- 0.3 h). LH pulses started and peaked later than the corresponding portal GnRH pulses (onset difference, 10 +/- 1 min; peak difference, 16 +/- 1 min; P < 0.01 for both), but the times of pulse onset and peak were not significantly different from those of concomitant CSF GnRH pulses (onset difference, 8 +/- 6 min; peak difference, 8 +/- 4 min). Although the times of pulse onset and peak did not differ between the portal and CSF GnRH compartments (onset difference, 4 +/- 6 min; peak difference, 6 +/- 2 min), CSF GnRH pulses were longer than their portal counterparts (CSF, 38 +/- 3 min; portal, 15 +/- 1 min; P < 0.01). The amplitude of jugular LH pulses was strongly correlated (r2 = 0.85) with portal GnRH pulse amplitude, but not with that of CSF GnRH pulses (r2 = 0.45); there was no correlation between portal and CSF GnRH pulse amplitudes (r2 = 0.25). These data show that third ventricular CSF GnRH reliably relates neurosecretory events occurring within the hypophyseal portal system at the time of the preovulatory LH surge, but is not as precise as portal GnRH in marking a LH pulse. PMID- 9348197 TI - Active repression by thyroid hormone receptor splicing variant alpha2 requires specific regulatory elements in the context of native triiodothyronine-regulated gene promoters. AB - Structural requirements for the inhibitory action of thyroid hormone receptor splicing variant alpha2 (TR alpha2) on T3/TRbeta1-mediated transactivation were investigated in native promoters of two T3-regulated genes: the brain-specific myelin basic protein (MBP) and the housekeeping malic enzyme (ME). T3/TRbeta1 transactivation of MBP256-chloramphenicol acetyl transferase (CAT) and ME315-CAT constructs was inhibited and unaffected by TR alpha2, respectively. In electrophoretic mobility shift assays, TR alpha2 bound MBP-thyroid response element (TRE) as a monomer but failed to interact with ME-TRE. Mutations of ME TRE allowed TR alpha2 binding but not inhibition of T3/TRbeta1-mediated transactivation. In the context of the MBP promoter, replacement of MBP-TRE with ME-TRE or exchange of MBP TATA-like box with the ME GC-rich region spanning the transcription start site abolished TR alpha2 dominant negative action. Simultaneous introduction of both MBP-TRE and MBP TATA-like box in the context of ME promoter, however, triggered TR alpha2 inhibition of T3/TRbeta1 transactivation, indicating that these regulatory elements are necessary, but not individually sufficient, to mediate TR alpha2 dominant negative activity. Functional studies at low TR alpha2/TRbeta1 ratios revealed that binding to TRE facilitates TR alpha2 dominant negative action while prevention of DNA interaction by altering TR alpha2 P-box structure preserved TR alpha2 inhibitory effect, although with lower potency. In conclusion, the results suggest that, in native promoters of T3-regulated genes, a dual molecular mechanism, with DNA binding dependent and DNA-binding independent components, underlies TR alpha2 dominant negative activity. PMID- 9348198 TI - Relaxin counteracts myocardial damage induced by ischemia-reperfusion in isolated guinea pig hearts: evidence for an involvement of nitric oxide. AB - Relaxin was previously shown to cause coronary vasodilation and to inhibit mast cell activation through a stimulation of endogenous nitric oxide production. This suggests that relaxin may have beneficial effects on ischemia-reperfusion-induced myocardial injury, which is triggered by endothelial damage and impaired nitric oxide generation. In this study, we tested the effect of relaxin on isolated and perfused guinea pig hearts subjected to ischemia and reperfusion. Ischemia was induced by ligature of the left anterior descending coronary artery; removal of the ligature induced reperfusion. Relaxin, at the concentration of 30 ng/ml of perfusion fluid, causes: a significant increase in coronary flow and in nitric oxide generation; a significant decrease in malonyldialdehyde production and in calcium overload, both markers of myocardial injury; an inhibition of mast cell granule exocytosis and histamine release, which are known to contribute to myocardial damage; a reduction of ultrastructural abnormalities of myocardial cells; an improvement of heart contractility. The beneficial effects of relaxin were blunted by the NO synthase inhibitor L-NMMA. The current study provides first experimental evidence that relaxin has a powerful protective effect on the heart undergoing ischemia and reperfusion acting through a nitric oxide-driven mechanism. PMID- 9348199 TI - Evidence for involvement of protein kinase C (PKC)-zeta and noninvolvement of diacylglycerol-sensitive PKCs in insulin-stimulated glucose transport in L6 myotubes. AB - We examined the question of whether insulin activates protein kinase C (PKC)-zeta in L6 myotubes, and the dependence of this activation on phosphatidylinositol (PI) 3-kinase. We also evaluated a number of issues that are relevant to the question of whether diacylglycerol (DAG)-dependent PKCs or DAG-insensitive PKCs, such as PKC-zeta, are more likely to play a role in insulin-stimulated glucose transport in L6 myotubes and other insulin-sensitive cell types. We found that insulin increased the enzyme activity of immunoprecipitable PKC-zeta in L6 myotubes, and this effect was blocked by PI 3-kinase inhibitors, wortmannin and LY294002; this suggested that PKC-zeta operates downstream of PI 3-kinase during insulin action. We also found that treatment of L6 myotubes with 5 microM tetradecanoyl phorbol-13-acetate (TPA) for 24 h led to 80-100% losses of all DAG dependent PKCs (alpha, beta1, beta2, delta, epsilon) and TPA-stimulated glucose transport (2-deoxyglucose uptake); in contrast, there was full retention of PKC zeta, as well as insulin-stimulated glucose transport and translocation of GLUT4 and GLUT1 to the plasma membrane. Unlike what has been reported in BC3H-1 myocytes, TPA treatment did not elicit increases in PKCbeta2 messenger RNA or protein in L6 myotubes, and selective retention of this PKC isoform could not explain the retention of insulin effects on glucose transport after prolonged TPA treatment. Of further interest, TPA acutely activated membrane-associated PI 3 kinase in L6 myotubes, and acute effects of TPA on glucose transport were inhibited, not only by the PKC inhibitor, LY379196, but also by both wortmannin and LY294002; this suggested that DAG-sensitive PKCs activate glucose transport through cross-talk with phosphatidylinositol (PI) 3-kinase, rather than directly through PKC. Also, the cell-permeable, myristoylated PKC-zeta pseudosubstrate inhibited insulin-stimulated glucose transport both in non-down-regulated and PKC depleted (TPA-treated) L6 myotubes; thus, the PKC-zeta pseudosubstrate appeared to inhibit a protein kinase that is required for insulin-stimulated glucose transport but is distinct from DAG-sensitive PKCs. In keeping with the latter dissociation of DAG-sensitive PKCs and insulin-stimulated glucose transport, LY379196, which inhibits PKC-beta (preferentially) and other DAG-sensitive PKCs at relatively low concentrations, inhibited insulin-stimulated glucose transport only at much higher concentrations, not only in L6 myotubes, but also in rat adipocytes, BC3H-1 myocytes, 3T3/L1 adipocytes and rat soleus muscles. Finally, stable and transient expression of a kinase-inactive PKC-zeta inhibited basal and insulin-stimulated glucose transport in L6 myotubes. Collectively, our findings suggest that, whereas PKC-zeta is a reasonable candidate to participate in insulin stimulation of glucose transport, DAG-sensitive PKCs are unlikely participants. PMID- 9348200 TI - Studies of melatonin effects on epithelia using the human embryonic kidney-293 (HEK-293) cell line. AB - The expression of melatonin receptors (MR) of the Mel1a subtype in basolateral membrane of guinea pig kidney proximal tubule suggests that melatonin plays a role in regulating epithelial functions. To investigate the cellular basis of melatonin action on epithelia, we sought to establish an appropriate in vitro culture model. Epithelial cell lines originating from kidneys of dog (MDCK), pig (LLC-PK1), opossum (OK), and human embryo (HEK-293) were each tested for the presence of MR using 2-[125I]iodomelatonin (125I-MEL) as a radioligand. The HEK 293 cell line exhibited the highest specific 125I-MEL binding. By intermediate filament characterization, the HEK-293 cells were determined to be of epithelial origin. Binding of 125I-MEL in HEK-293 cells demonstrated saturability, reversibility, and high specificity with an equilibrium dissociation constant (Kd) value of 23.8 +/- 0.5 pM and a maximum number of binding sites (Bmax) value of 1.17 +/- 0.11 fmol/mg protein (n = 5), which are comparable with the reported Kd and Bmax values in human kidney cortex. Coincubation with GTPgammaS (10 microM) and pertussis toxin (100 ng/ml) provoked a marked decrease in binding affinity (Kd was increased by a factor of 1.5-2.0), with no significant difference in Bmax. Melatonin (1 microM) decreased the forskolin (10 microM) stimulated cAMP level by 50%. HEK-293 cells do not express dopamine D1A receptor. Following transient transfection of HEK-293 cells with human dopamine D1A receptor (hD1A-R), exposure of the cells to dopamine stimulated an increase in the level of cAMP. Similarly, transient transfection of HEK-293 cells with rat glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic peptide (GIP), and PTH type 1 receptors, each resulted in an hormone inducible increase in cAMP levels. Surprisingly, only the stimulatory effect of dopamine could be inhibited by exposure to melatonin. The inhibitory effect of melatonin on dopamine D1 induced increase in cAMP was completely inhibited by pertussis toxin (100 ng/ml, 18 h). Immunoblot and immunocytochemical studies were carried out using two polyclonal antibodies raised against the extra and cytoplasmic domains of Mel1a receptor. Immunoblot studies using antibody against the cytoplasmic domain of Mel1a receptor confirmed the presence of a peptide blockable 37 kDa band in HEK 293 cells. Indirect immunofluorescent studies with both antibodies revealed staining predominantly at the cell surface, but staining with the antibody directed against the cytoplasmic domain required prior cell permeabilization. By RT-PCR, HEK-293 cells express both Mel1a and Mel1b messenger RNAs, but the messenger RNA level for Mel1b is several orders of magnitude lower than for Mel1a. We conclude that HEK-293 cells express MR predominantly of the Mel1a subtype. Our evidence suggests that one of the ways that melatonin exerts its biological function is through modulation of cellular dopaminergic responses. PMID- 9348201 TI - Brain region-specific regulation of luteinizing hormone-releasing hormone messenger ribonucleic acid in the male ferret: interactions between pubertal maturation and testosterone. AB - This study examined the regulation of LHRH messenger RNA (mRNA) during pubertal maturation and by testosterone in male ferrets. Prepubertal and postpubertal ferrets were either intact or were castrated and treated with daily injections of oil or 5 mg/kg testosterone propionate for 14 days. In situ hybridization for LHRH mRNA was performed using an 35S-labeled 48-base oligonucleotide complementary to the human LHRH-coding region. Computerized image analysis was performed on cells in the preoptic area, retrochiasmatic area, arcuate nucleus (ARC), and median eminence; cells were classified as labeled if the number of pixels representing silver grains over the cell was 5 or more times the number of background silver grain pixels. Both pubertal maturation of intact males and castration of prepubertal males resulted in an increase in the number of labeled cells in the ARC. These effects were not observed in any of the other three brain regions, suggesting that ARC LHRH-producing neurons are of primary importance in the presumed increase in LHRH release that occurs as a consequence of either pubertal maturation or castration of prepubertal males. Castration of adults did not increase the number of labeled cells in any brain area, but resulted in an increase in silver grains per labeled cell only in the preoptic area. Thus, LHRH mRNA is regulated during puberty primarily in the ARC, and the particular cell group in which LHRH mRNA is most strongly regulated by testosterone changes with pubertal maturation. PMID- 9348202 TI - Differential hormonal regulation of vascular endothelial growth factors VEGF, VEGF-B, and VEGF-C messenger ribonucleic acid levels in cultured human granulosa luteal cells. AB - The development of ovarian follicles and subsequent corpus luteum formation is accompanied by very active angiogenesis. Ovarian granulosa cells produce vascular endothelial growth factor (VEGF), which is a potent endothelial cell mitogen and an angiogenic agent. The complementary DNAs of two other factors structurally related to VEGF, namely VEGF-B and VEGF-C, were recently cloned, but little is known of their regulation in the ovary. We first studied the expression of the messenger RNAs (mRNAs) of the three VEGF isotypes in freshly isolated human granulosa-luteal (GL) cells obtained at oocyte retrieval for in vitro fertilization. The hormonal regulation of these mRNAs was subsequently studied in primary cultures of human GL cells. Analysis of cultured GL cell RNA by reverse transcription-PCR revealed that these cells express the alternatively spliced transcripts representing 121-, 145-, and 165-amino acid VEGF isoforms. Northern blot hybridization analyses indicated that transcripts of 4.5 and 3.7 kilobases for VEGF, and 1.4 and 2.4 kilobases for VEGF-B and VEGF-C, respectively, are expressed in human GL cells. The basal VEGF mRNA levels declined steadily, whereas VEGF-B mRNA levels were rather invariant over a 10-day culture period of GL cells. In contrast, VEGF-C mRNA levels increased toward the end of culture. For studying the hormonal regulation of VEGF isotype mRNAs, GL cells were treated with hCG, recombinant human FSH, PGE2, as well as 8-bromo-cAMP and 12-O tetradecanoylphorbol 13-acetate, which activate protein kinase A- and protein kinase C-dependent signaling pathways, respectively. All test agents stimulated the expression of VEGF mRNA levels in a concentration-dependent manner. Time course studies indicated that all treatments induced VEGF mRNA levels as early as incubation for 2 h, and the effect was sustained up to 48 h. VEGF-B mRNA levels were not regulated by any of the test agents. However, we found that hCG and 8 bromo-cAMP decreased VEGF-C mRNA levels with a maximal response observed at 24 and 48 h after cellular treatment. We conclude that the mRNAs of VEGF, VEGF-B, and VEGF-C are expressed in human GL cells and that their mRNA steady state levels are regulated in cultured human GL cells in an isotype-specific manner. The differential regulation of VEGF, VEGF-B, and VEGF-C in human GL cells suggests that distinct VEGF isotypes may play different roles during the vascularization of the human ovarian follicle and corpus luteum. PMID- 9348203 TI - Molecular cloning of ovine and bovine type I interferon receptor subunits from uteri, and endometrial expression of messenger ribonucleic acid for ovine receptors during the estrous cycle and pregnancy. AB - Interferon-tau (IFN-tau), a type I IFN structurally related to IFN-alpha, is regarded as the major antiluteolytic factor secreted by the conceptus of ruminant ungulate species before definitive trophoblast attachment and implantation. It mediates its effects by acting on the uterine endometrium, where it blunts the normal pulsatile production of PGF2alpha, presumably as a result of its binding to type I IFN receptors. In this study, we describe the complementary DNAs for the two known subunits, IFNAR1 and IFNAR2, of this receptor isolated from bovine and ovine endometrial complementary DNA libraries by homology cloning. Although there is extensive inferred amino acid sequence similarity between bovine and ovine IFNAR1 (92% identity) and between bovine and ovine IFNAR2 (88% identity), they have diverged extensively from the human receptor subunits (approximately 67% and approximately 58% identity, respectively). Despite these differences in primary structure, the respective subunits from all three species are organized similarly in their extracellular and cytoplasmic regions, and the bovine and ovine subunits have each retained a number of polypeptide motifs implicated in signal transduction. These uterine receptors also appear not to be splice variants. The cloned ovine IFNAR1 subunit, for example, possesses the expected four extracellular SD100 domains of full-length bovine and huIFNAR1, and only the homologs of the so-called long form (huIFNAR2c) of human IFNAR2 have so far been identified. RT-PCR procedures indicate that the messenger RNA for both subunits are found, not only in endometrium, but in all other tissues examined except those ofpreimplantation conceptuses, which presumably cannot respond to the IFN tau they produce. Quantitative RNase protection assays of ovine endometrial RNA show that the expression of neither subunit changes greatly during the estrous cycle or pregnancy. These data suggest that the type I IFN receptor, which is expressed by the endometrium and binds IFN-tau, is probably not a structurally unusual form. PMID- 9348204 TI - Expression of steroid sulfatase during embryogenesis. AB - Neurosteroids are steroids that are synthesized de novo in the brain from cholesterol and, in general, mediate their effects through ion-gated channel receptors such as gamma-aminobutyric acidA (GABA[A]) and N-methyl-D-aspartate receptors rather than through classical nuclear steroid hormone receptors. Steroid hormones are known to exist not only as free compounds, but also as sulfated derivatives. Pharmacological studies indicate that unconjugated and sulfated steroids, such as pregnenolone and pregnenolone sulfate, may have opposite effects on GABA(A) receptors. Thus, pregnenolone acts as a potent positive allosteric modulator of gamma-aminobutyric acid action at GABA(A )receptors, whereas pregnenolone sulfate acts as a potent negative modulator. Recent experiments also suggest that dehydroepiandrosterone and dehydroepiandrosterone sulfate may have distinct effects on growth of neurites from embryonic neocortical neurons in vitro. Thus, regulation of steroid sulfation may have profound behavioral and morphological effects on the nervous system. We, therefore, studied the developmental expression of the enzyme steroid sulfatase (STS), which converts sulfated steroids to free steroids. By in situ hybridization, STS messenger RNA was expressed in the embryonic mouse cortex, hindbrain, and thalamus during the last third of gestation. The sites of expression of STS were similar to those of P450c17, suggesting that these two enzymes may have concerted actions in similar functional processes. PMID- 9348205 TI - Regulation of corticotropin-releasing factor (CRF) messenger ribonucleic acid and CRF peptide in the amygdala: studies in primary amygdalar cultures. AB - Amygdalar CRF has been implicated in the mediation of stress behaviors. The signal transduction pathways that regulate amygdalar CRF are not well understood. In this report, we have examined the effect of protein kinase A and C activators, dexamethasone, and interleukin 6 on CRF messenger RNA (mRNA) and CRF peptide expression in dissociated amygdalar cultures. The amygdala from E19 rat pups was dissected out bilaterally and dissociated in 0.25% trypsin for 10-15 min and plated. On day 17 in culture, CRF mRNA and peptide were measured following treatment with the following agents: forskolin, the phorbol ester-phorbol 12 myristate 13-acetate (TPA), dexamethasone, and interleukin-6 (IL6). Both forskolin and IL6, but not TPA, increased CRF mRNA in a time- and dose-dependent manner. Secretion and intracellular content of the CRF peptide also increased with both forskolin and IL6 treatment but not with TPA. Dexamethasone treatment did not alter the expression of CRF message or peptide. Transfection of the primary cultures with a rat CRF promoter-luciferase reporter construct followed by treatment with all four agents produced alterations in luciferase expression that were consistent with changes observed at the level of CRF mRNA and peptide. The results suggest that CRF regulation in the amygdala differs from that known to occur in the hypothalamus, and that elevation of IL6 levels within the central nervous system may directly act to stimulate CRF production and secretion from limbic structures such as the amygdala, to promote subsequent behavioral changes. PMID- 9348206 TI - Stimulating effect of both human recombinant inhibin A and activin A on immature porcine Leydig cell functions in vitro. AB - In addition to the regulation of FSH secretion, it has been clearly shown that inhibin and activin have paracrine/autocrine effects in the gonads. We have studied the effect of human recombinant inhibin A and human recombinant activin A on immature porcine Leydig cells in vitro. Leydig cells were prepared by collagenase digestion of testes from 3-week-old piglets, purified on Percoll gradient, then cultured in a chemically defined medium. The cells were treated with increasing amounts of inhibin A or activin A (0.5-200 ng/ml). Direct application of either inhibin A or activin A on Leydig cells for 4 or 48 h did not stimulate basal testosterone secretion. Conversely, treatment of the cells for 48 h with either factor resulted in a dose-dependent increase in hCG stimulated testosterone secretion (10[-9] M hCG, 2 h) with a maximal effect of 2.40 +/- 0.37- and 2.43 +/- 0.37-fold increases for inhibin A and activin A, respectively, and these changes were associated with a slight increase in LH/hCG binding sites (1.37 +/- 0.19- and 1.24 +/- 0.11-fold increases). In addition, both inhibin A and activin A enhanced messenger RNA (mRNA) levels of LH/hCG receptor (2.75 +/- 0.40- and 2.53 +/- 0.60-fold increases) and cytochrome P450 17alpha-hydroxylase (6 +/- 1- and 3.5 +/- 0.6-fold increases), but had no effect on side-chain cleavage cytochrome P450 or cytochrome P450 aromatase mRNAs. 3beta Hydroxysteroid dehydrogenase mRNA levels were increased (3.1 +/- 1.3-fold increase) by activin A, but not by inhibin A. However, inhibin A blocked the stimulatory action of activin A. In keeping with these changes in the steroidogenic enzyme mRNAs, both peptides enhanced the conversion of exogenous 22R-hydroxycholesterol and progesterone, but only activin A increased the conversion of dehydroepiandrosterone into testosterone. In conclusion, our findings demonstrate that both inhibin A and activin A have a stimulatory effect on immature porcine Leydig cell differentiated function in vitro. As inhibin has a stimulatory and activin has an inhibitory effect on rat Leydig cell function in vitro, the effects of these factors on Leydig cells seem to be species dependent. PMID- 9348207 TI - Maternal adrenalectomy eliminates a surge of plasma dehydroepiandrosterone in the mother and attenuates the prenatal testosterone surge in the male fetus. AB - Previous work has established a number of sex-related deficits in immune function, behavior, and endocrine responses to stress in the offspring of dams exposed to ethanol. To examine the potential role of maternal glucocorticoids as a mediator of these sexually dimorphic effects in the fetus, we examined the influence of prenatal alcohol exposure in the presence or absence of maternal glucocorticoids on fetal plasma corticosterone (CORT) production. An additional question to be addressed by these studies was whether maternal adrenalectomy could eliminate the known inhibition by ethanol of the prenatal surge of plasma testosterone in male fetuses. Pregnant dams were adrenalectomized (ADX) or sham adrenalectomized on gestational day (G) 7 and placed on a liquid diet containing 35% ethanol-derived calories or pair-fed an isocaloric control diet throughout the experiment. On G18, G19, and G21, plasma levels of CORT, testosterone, and dehydroepiandrosterone (DHEA) were measured in male and female fetuses and their mothers. Ethanol administration consistently increased maternal plasma CORT levels but did not significantly alter CORT levels in the fetus. Maternal ADX resulted in compensatory increases in fetal CORT levels that were lower in fetuses of ADX dams on alcohol, suggesting a direct effect of ethanol on fetal pituitary-adrenal activity. There were no significant sex differences in fetal plasma CORT levels in response to any of these manipulations. A novel surge of maternal plasma DHEA was found on G19 that was absent in plasma from ADX dams. In spite of the absence of a surge on G19, plasma DHEA levels of ADX dams rose from very low levels at G18 to levels on G21 that were significantly higher than in Sham dams. A normal testosterone surge was observed in male fetuses on G18 and G19 from sham-adrenalectomized dams administered the pair-fed diet. However, this surge was greatly attenuated in males administered ethanol and also in male fetuses from ADX dams. These results reveal a direct inhibitory influence of ethanol on fetal CORT secretion as well as on the prenatal testosterone surge in males. Furthermore, these studies demonstrate the presence of a surge of DHEA in the pregnant rat. Overall, these data suggest that there is a critical adrenal factor in the rat that regulates the maternal surge of DHEA on G19 and the prenatal testosterone surge of male fetuses on G18-19. PMID- 9348209 TI - Corticotropin-releasing hormone (CRH) inhibits steroid biosynthesis by cultured human granulosa-lutein cells in a CRH and interleukin-1 receptor-mediated fashion. AB - The presence of immunoreactive CRH was recently demonstrated in human ovaries. CRH immunoreactivity was localized by immunohistochemistry in the cytoplasm of thecal cells surrounding the ovarian follicles, in luteinized cells of the stroma, and in large granulosa-derived luteinized cells of developing corpora lutea. Also, CRH and its receptors were identified in Leydig cells of the testis where CRH was shown to inhibit testosterone biosynthesis. To examine the role of CRH in the ovary, we studied its effect on estradiol (E2) and progesterone (P4) release by human granulosa cells obtained from women undergoing in vitro fertilization for male factor infertility or uni- or bilateral tubal impatency. In all subjects, superovulation was induced by treatment with gonadotropins. The effects of graded doses of ovine CRH (10[-11]-10[-6] mol/liter) were evaluated in the conditioned medium obtained after 24 h incubation of the cells. All CRH concentrations employed except for the lowest one (10[-11] mol/liter) caused a significant decrease of media E2 and P4 levels. Maximal inhibition for both E2 and P4 production was obtained by 10[-6] mol/liter CRH concentration, which decreased hormone production by 39% and 34%, respectively. The alpha-helical CRH9 41 antagonist at 10(-6) and 10(-7) mol/liter blocked the suppressive effect of 10(-9) mol/liter CRH on both E2 and P4 secretion, while it had no effect when added to the culture media without CRH. Since interleukin (IL-1)-1 mediates certain actions of CRH on leukocytes, we examined whether the CRH effect on ovarian steroidogenesis was IL-1-mediated. Interleukin-1 receptor antagonist at 10(-7) and 10(-6) mol/liter blocked the inhibitory effects of CRH on E2 and P4 secretion, while it had no effect in the absence of CRH. In conclusion, CRH exerts a CRH- and IL-1 receptor-mediated inhibitory effect on ovarian steroidogenesis and might be actively involved in the still enigmatic processes of follicular atresia and luteolysis. PMID- 9348208 TI - Cellular mechanisms involved during oxytocin-induced prostaglandin F2alpha production in endometrial epithelial cells in vitro: role of cyclooxygenase-2. AB - PGs are important regulators of reproductive processes. At the time ofluteolysis in vivo, PGF2alpha is produced by endometrial cells, in response to oxytocin (OT). The mechanism by which OT induces the release of PGF2alpha remains to be defined. We have used 13 different cultures of bovine epithelial endometrial cells to study the effect of OT on the regulation of PGF2alpha and to identify the possible involvement of cyclooxygenases (COXs). OT induced a dose-dependent increase of both inositol phosphates (IPs) and [Ca2+]i concentration in epithelial cells labeled with [3H]-myoinositol or loaded with fura-2 (using a fluorescent microscope imaging system), respectively. OT induced a dose-dependent increase of both PGF2alpha production and COX-2 gene expression (as demonstrated by RT-PCR and Northern blots). PGF2alpha production was increased from 13.3 +/- 2.0 to 166.8 +/- 22.5 ng/ml (P < 0.0001). On the other hand, COX-2/beta-actin mRNA gene expression (as determined by densitometric analysis) was increased 5.1 +/- 0.7-fold (P < 0.001) with OT (10[-7] M) treatment, compared with control. Addition of indomethacin (1 microM) and a specific COX-2 inhibitor (NS-398, 1 microM) blocked the OT-induced PGF2alpha production. COX-1 and phospholipase A2 mRNA were expressed at steady-state levels, but no effect of OT was detected on their regulation. Combined to OT, 10 microq/ml of recombinant ovine interferon tau (roIFN-tau) was able to decrease significantly (P < 0.0001) the dose dependent increase of PGF2alpha production. Furthermore, partial bovine COX-1 (777 pb) and COX-2 (449 bp) cDNAs were cloned and sequenced. An homology of 83% and 97% was found in relation with rat and sheep, for COX-1, respectively. COX-2 was found to bear 84%, 86%, and 87% of homology in relation to rat, guinea pig, and human, respectively. Collectively, these results demonstrate, for the first time, that COX-2 is involved in the mechanism by which OT regulates PGF2alpha production in the endometrium. PMID- 9348210 TI - The different forms of the prolactin receptor in the rat corpus luteum: developmental expression and hormonal regulation in pregnancy. AB - The corpora lutea of pregnancy in the rat are highly dependent on the action of PRL and PRL-like hormones to hypertrophy and to produce progesterone needed for the maintenance of gestation. Two forms of the PRL receptor (PRL-R), designated as long (PRL-RL) and short (PRL-RS), have been described in rat tissues. To determine whether both forms are present in the corpus luteum during pregnancy and to examine the developmental and hormonal regulation of their expression, total RNA isolated from corpora lutea at different stages of pregnancy and from highly luteinized granulosa cells subjected to different hormonal treatments were analyzed by semiquantitative RT-PCR. Immunoblotting of luteal proteins from early and late pregnancy was also performed to determine if the pattern of PRL-R proteins follows that of PRL-R messenger RNA (mRNA) expression. In addition, the correlation between the well characterized PRL-regulated gene, 20alpha hydroxysteroid dehydrogenase (20alpha-HSD), and PRL-R gene expression was investigated during the time of luteolysis. Both PRL-RL and PRL-RS mRNA and protein were expressed in corpora lutea of pregnancy, with the long form being the most dominant at all stages. Whereas no changes in mRNA level of either PRL RL or PRL-RS were found until day 20 of gestation, a profound decline in PRL-R mRNA and protein for both receptor types occurred at the end of pregnancy. This drop in PRL-R expression was accompanied by a sharp and abrupt expression of 20alpha-HSD mRNA. Studies performed in vivo and in luteinized cells in culture indicate that PRL can up-regulate the expression of the PRL-RL mRNA, an effect prevented by the tyrosine kinase inhibitor, genistein. PRL-RL mRNA was also selectively increased by cAMP. In summary, the results of this investigation have established that: 1) the corpus luteum of pregnancy expresses both the short and long forms of the PRL-R with the long form being more abundant; 2) the mRNA for both forms of the PRL-R remains at constant levels throughout pregnancy but drops before parturition; 3) the decline in PRL-R mRNA at the end of pregnancy is accompanied by a dramatic rise in 20alpha-HSD; 4) PRL is able to increase the expression of PRL-R mRNA; and that 5) both A kinase and tyrosine kinase mediated pathways appear to participate in the up-regulatory mechanism involved in PRL-R mRNA expression. PMID- 9348211 TI - Vanadate, but not insulin, inhibits insulin receptor gene expression in rat hepatoma cells. AB - Insulin and vanadate treatments have recently been shown to reverse the overexpression of the hepatic insulin receptor (IR) gene in streptozotocin induced diabetic rats. To better understand the mechanisms underlying these effects, the abilities of insulin and vanadate to affect IR gene expression have been comparatively examined in Fao hepatoma cells, an insulin-responsive cell line. Exposure of Fao cells to insulin (1 microM) or vanadate (500 microM) for 24 h led to a 2-fold decrease in IR number in total cellular membranes. Insulin treatment did not affect IR messenger RNA (mRNA) level regardless of time of exposure and concentration. In contrast, vanadate treatment caused a time- and dose-dependent decrease in IR mRNA level, which was maximal (4-fold change) after a 24-h exposure to 500 microM vanadate and was fully reversible. Insulin treatment increased from 28 to 39% the relative expression of isotype A IR mRNA, but vanadate treatment did not significantly affect this parameter. Vanadate treatment did not modify mRNA half-life (3.5 h) in 5, 6 dichlorobenzimidazole riboside-treated cells but decreased by 4-fold the transcriptional activity of the IR gene. These data show for the first time that, although both insulin and vanadate decrease total cellular IR number in Fao cells, only vanadate decreases IR mRNA level. It does so by inhibiting transcription of the IR gene, suggesting an action on the gene promoter which could be mediated by changes in the level of expression and/or of phosphorylation of trans-acting factors. PMID- 9348212 TI - Clearance of 125I-labeled interleukin-6 from brain into blood following intracerebroventricular injection in rats. AB - To test the hypothesis that interleukin-6 (IL-6) induced within the brain can be released into peripheral blood, 125I-labeled IL-6 was injected into the lateral cerebral ventricle of rats, and its concentration in peripheral blood followed serially. Acid-precipitable tracer appeared within 5 min of injection and entered the blood following first-order kinetics (fractional rate, 0.0116 +/- 0.0022/min). Comparison of areas under the curve of intracerebroventricular (icv) vs. iv injection showed that 37.1-46.5% of tracer injected into the lateral cerebral ventricle appeared in the blood over a 4-h period. icv IL-6 exits at least in part via venous drainage (superior sagittal sinus/aortic concentration gradient was 1.47 +/- 0.23 and 3.05 +/- 0.87 in two separate groups). Prior icv injection of human IL-1beta (100 ng) did not alter rate of degradation or of exit ofradioiodine-labeled IL-6 from the brain. These studies indicate that a relatively high proportion of IL-6 that arises in the brain enters the peripheral circulation. Direct secretion of IL-6 from brain to blood may be a mechanism by which the brain modifies peripheral metabolic, endocrine, and immune activity. PMID- 9348213 TI - Circulating and tissue forms of the intestinal growth factor, glucagon-like peptide-2. AB - Glucagon-like peptide-2 (GLP-2) has recently been identified as a stimulator of intestinal epithelial growth, prompting the development of RIA and HPLC methodologies to study this peptide in more detail. A GLP-2-specific antiserum (UTTH-7) was developed that recognizes amino acids 25-30 of human and rat GLP-2 (1-33). UTTH-7 cross-reacts with N- and C-terminally modified forms of GLP-2, proglucagon, and the major proglucagon fragment. Analysis of rat ileal extracts demonstrated the presence of GLP-2-(1-33) as well as significant amounts of GLP-2 (3-33) (16 +/- 7% of total GLP-2). The level of total immunoreactive GLP-2 in plasma from fasted rats was 700 +/- 71 pg/ml, and this increased 3.6-fold (P < 0.001) in 24-h fed rats. HPLC analysis demonstrated the presence of both GLP-2-(1 33) and GLP-2-(3-33) in plasma from fasted rats, with increments in both peptides in plasma from fed rats. Immunoreactive GLP-2 increased in plasma from human subjects 2 h after a meal, rising from 851 +/- 230 to 1106 +/- 211 pg/ml (P < 0.05); 15 +/- 4% of this immunoreactivity was accounted for by the presence of intact GLP-2. HPLC showed the presence of both GLP-2-(1-33) and GLP-2-(3-33) in plasma from fed humans. Incubation of human GLP-2-(1-33) with the enzyme dipeptidylpeptidase IV resulted in liberation of GLP-2-(3-33), whereas replacement of Ala2 with Gly2 prevented this cleavage. Thus, while GLP-2-(1-33) is a major circulating and tissue form of GLP-2, GLP-2-(3-33) is a significant component ofimmunoreactive GLP-2 in both intestine and plasma. PMID- 9348214 TI - Expression of functional luteinizing hormone (LH) receptor and its messenger ribonucleic acid in bovine uterine veins: LH induction of cyclooxygenase and augmentation of prostaglandin production in bovine uterine veins. AB - We have previously reported that bovine endometrium contains LH/human CG binding receptors and LH induces cyclooxygenase and prostaglandin production in the bovine endometrium. The present study investigated 1) whether bovine uterine vein and artery contain LH receptor messenger RNA (mRNA) and receptor protein and 2) whether LH can regulate the formation of vasoactive eicosanoids by the uterine vein. The uterine vein endothelium, but not the uterine artery, contained LH receptor mRNA transcript essentially identical to that found in the bovine corpus luteum. The uterine vein endothelium also contained a 95-kDa immunoreactive receptor protein that bound to rat anti-LH receptor antibody in Western blots. The LH receptor mRNA and LH receptor were maximally expressed in the uterine vein from cows in proestrus/estrus compared with cows in luteal or postovulatory phases. Incubation of endothelial minces of uterine vein with LH resulted in a 2 fold increase in cyclooxygenase concentration as determined by Western blot using an antibody to ram seminal vesicle cyclooxygenase. The increase in cyclooxygenase was maximal in cows in proestrus/estrus compared with postovulatory and luteal phase cows. Incubation of proestrous/estrous uterine vein or artery minces with LH or mellitin (a phospholipase A2 stimulator) caused increased production of eicosanoids. In the uterine vein, LH caused a significant increase in both PGF2alpha (basal 4.1 +/- 0.4 vs. 5.7 +/- 0.4 ng/100 mg x 6 h, P < 0.01; N = 9 cows) and PGE2 (basal 5.7 +/- 0.3 vs. 7.7 +/- 0.8 ng/100 mg x 6 h, P < 0.01; N = 6 cows) but had no effect on prostaglandin production by the artery. Mellitin increased PGF2alpha production by both uterine vein and artery minces but had no effect on PGE2 production in either tissue. Addition of steroids (progesterone, estradiol) or cytokines (tumor necrosis factor-alpha, IL-6) to the uterine vascular tissues had essentially no effect on prostanoid production. In summary, bovine uterine vein from proestrous/estrous cows expressed the LH receptor and its mRNA. Expression of the receptor may have physiological significance as LH induces cyclooxygenase and increases prostaglandin release in the uterine vein. The maximal stimulation of the receptor and its mRNA at proestrus/ estrus may serve to increase the amounts of prostanoids reaching the regressing corpus luteum either directly by increasing prostanoid production or indirectly by increasing the blood flow to the ovary. PMID- 9348215 TI - Induction of carbonic anhydrase II expression in osteoclast progenitors requires physical contact with stromal cells. AB - Carbonic anhydrase II (CA II) expression is vital to normal osteoclast function. We and others have previously reported induction of CA II messenger RNA (mRNA) expression by 1,25(OH)2D3 in myelomonocytic cells and marrow culture. However, since 1,25(OH)2D3 stimulates osteoclast differentiation as well, we wished to separate direct effects of 1,25(OH)2D3 on the CA II gene from the differentiating effects of the hormone. Using primary murine mixed marrow cultures, we measured CA II mRNA expression by RT-PCR. 10 nM 1,25(OH)2D3 dose dependently induced expression of CA II mRNA (4.12 +/- 0.68-fold) at day 4 in culture compared with control with an ED50 of 0.25 nM. When nonadherent marrow cells containing osteoclast progenitors were depleted of stromal cells and exposed to 10 nM 1,25(OH)2D3, CA II mRNA expression was decreased by more than 60%. Coculture of progenitors with ST-2 stromal cells for 3 days with 10 nM 1,25(OH)2D3 stimulated CA II expression by 22 +/- 3.6-fold. 1,25(OH)2D3 stimulated CA II mRNA expression in progenitors separated from ST-2 cells by transwells was insignificant demonstrating that the two cell types must be in physical contact. PTH also stimulated CA II mRNA expression (4.91 +/- 0.01-fold) to a similar degree as seen with 1,25(OH)2D3 treatment. These results demonstrate that induction of CA II in osteoclast progenitors requires their physical communication with stromal cells and is inseparable from the osteoclast differentiation process. PMID- 9348216 TI - Insulin-like growth factor binding protein-2 binds to cell surface proteoglycans in the rat brain olfactory bulb. AB - A family of six insulin-like growth factor binding proteins (IGFBPs) bind IGF-I and modulate its biological activity. IGFBPs may bind to macromolecules on the cell surface or pericellular extracellular matrix, and this interaction may modulate their effect on IGF activity. To date, little is known about the specificity of IGFBPs in the regulation of IGF action in the brain. We therefore explored whether IGFBPs were associated with cell membrane or extracellular matrix components in the rat brain. IGF-I binding sites with the characteristics of an IGFBP were found in the olfactory bulb mitral cell layer. This IGFBP was identified as IGFBP-2 by immunoprecipitation of both solubilized membrane preparations and cross-linked 125I-IGF: IGFBP complexes. While binding of IGFBP-2 to cell membranes was unaffected by RGD-containing peptide, it was inhibited by high salt concentration, suggesting interaction with proteoglycans. IGFBP-2 bound in vitro to the glycosaminoglycans chondroitin-4 and -6-sulfate, keratan sulfate, and heparin. IGFBP-2 also bound the proteoglycan aggrecan, an effect reduced by digestion of its glycosaminoglycans. Binding of IGFBP-2 to chondroitin-6-sulfate decreased the binding affinity of IGFBP-2 for IGF-I approximately 3-fold. Finally, an IGFBP-2 antibody coimmunoprecipitated IGFBP-2 and an approximately 200 kDa proteoglycan containing chondroitin-sulfate side chains from the rat olfactory bulb, providing definitive evidence for IGFBP-2 binding to olfactory bulb proteoglycans. These findings indicate that IGFBP-2 binds to proteoglycans in cell membranes of the rat olfactory bulb. Because we have previously shown that IGFs are highly expressed in the rat olfactory bulb, cell associated IGFBP-2 may have an important role in directing IGFs to specific sites in this brain region. PMID- 9348217 TI - Preservation of functioning human thyroid "organoids" in the scid mouse. IV. In vivo selection of an intrathyroidal T cell receptor repertoire. AB - To study the in vivo influence of thyroid cells on the T cell receptor repertoire in human autoimmune thyroid disease, we mixed lymphocyte-free thyrocytes (approximately 1.2 x 10[6]) from patients with Graves' disease with autologous peripheral blood mononuclear cells (PBMC; approximately 1.5 x 10[6]) and transplanted this mixture sc into scid mice while suspended in a basement membrane gel (approximately 0.4 ml). Controls included mice that received either thyrocytes only or PBMC only. The resulting artificial mixed cell thyroid organoids were explanted after 5 weeks, and their T cell receptor repertoire was examined. Of a total of 63 organoids constructed, 60 were recovered (95.2%). Total RNA was extracted and then analyzed by reverse transcription-PCR primarily for human T cell receptor (hTcR) Vbeta gene expression using 21 hTcR Vbeta amplimers. A restricted pattern of hTcR Vbeta gene expression was found, with 6 Vbeta genes (Vbeta5, 6, 7, 8, 13.1, and 18) predominantly expressed [P < 0.05, by ANOVA on ranks and Student-Newman-Keul's (SNK) test]. PBMC and control organoids showed no preferential selection of particular hTcR V gene-expressing T cells. This reductionist, mixed cell, thyroid model reflected earlier observations in human and murine autoimmune thyroid diseases in which a bias in hTcR V gene family expression had been observed. The model permitted in vivo T cell selection and/or enrichment of potentially disease relevant human T cells. PMID- 9348218 TI - Expression of different 17beta-hydroxysteroid dehydrogenase types and their activities in human prostate cancer cells. AB - The 17beta-hydroxysteroid dehydrogenase (17betaHSD) enzyme system governs important redox reactions at the C17 position of steroid hormones. Different 17betaHSD types (no. 1-4) have been identified to date in peripheral human tissues, such as placenta, testis, and breast. However, there is little information on their expression and activity in either normal or malignant prostate. In the present work, we have inspected pathways of 17beta-oxidation of either androgen or estrogen in human prostate cancer cells (LNCaP, DU145, and PC3) in relation to the expression of messenger RNAs (mRNAs) for 17betaHSD types 1-4. These cell systems feature distinct steroid receptor status and response to hormones. We report here that high expression levels of 17betaHSD4 were consistently observed in all three cell lines, whereas even greater amounts of 17betaHSD2 mRNA were detected solely in PC3 cells. Neither 17betaHSD1 nor 17betaHSD3 mRNAs could be detected in any cell line. From a metabolic standpoint, intact cell analysis showed a much lower extent of 17beta-oxidation of both androgen [testosterone (T)] and estrogen [estradiol (E2)] in LNCaP and DU145 cells compared to PC3 cells, where a greater precursor degradation and higher formation rates of oxidized derivatives (respectively, androstenedione and estrone) were observed. Using subcellular fractionation, we have been able to differentiate among 17betaHSD types 1-4 on the basis of their distinct substrate specificities and subcellular localization. This latter approach gave rise to equivalent results. PC3 cells, in fact, displayed a high level of microsomal activity with a low E2/T activity ratio and approximately equal apparent Km values for E2 and T, suggesting the presence of 17betaHSD2. Dehydrogenase specific activity with both E2 and T was also detected, although at lower levels, in LNCaP and DU145 cells. No evidence for reductase activity could be obtained in either the soluble or microsomal fraction of any cell line. As comparable expression levels of 17betaHSD4 were seen in the three cell lines, 17betaHSD2 is a likely candidate to account for the predominant oxidative activity in PC3 cells, whereas 17betaHSD4 may account for the lower extent of E2 oxidation seen in both LNCaP and DU145 cells. This is the first report on the expression of four different 17betaHSD types in human prostate cancer cells. It ought to be emphasized that for the first time, analysis of different 17betaHSD activities in either intact or fractionated cells harmonizes with the expression of relevant mRNAs species. PMID- 9348219 TI - Beta-cell lines derived from transgenic Cpe(fat)/Cpe(fat) mice are defective in carboxypeptidase E and proinsulin processing. AB - A spontaneous point mutation in the coding region of the carboxypeptidase E (CPE) gene in Cpe(fat)/Cpe(fat) mice affects proinsulin processing. Cell lines derived from the pancreatic beta-cells of Cpe(fat)/Cpe(fat) mice were generated by crossing C57BLKS/J-Cpe(fat)/+ mice with NOD mice expressing the simian virus 40 large T oncogene under the control of the rat insulin II promoter. Two cell lines, designated NIT-2 and NIT-3, were cultured from adenomatous islets obtained from F2 littermates and were compared with the NIT-1 cell line previously developed from mice with wild-type CPE. Electron microscopy of the cultured NIT-2 and -3 cells showed increased numbers of enlarged and electron-lucent granules compared with NIT-1 cells. Pro-CPE, but not the mature form of CPE, is present in NIT-2 and -3 cells, and neither pro-CPE nor CPE are secreted into the medium. Immunocytochemistry shows the pro-CPE to be localized to an endoplasmic reticulum like structure in NIT-3 cells. Proinsulin is less extensively processed in NIT-2 and -3 cells than in NIT-1 cells, indicating that the Cpe(fat) mutation affects both the endopeptidase and carboxypeptidase reactions. The secretion of insulin/proinsulin from NIT-2 and -3 cells is significantly elevated by secretagogues, indicating that CPE is not required for sorting proinsulin into the regulated pathway. PMID- 9348220 TI - Inhibition of transcription affects synthesis of steroidogenic acute regulatory protein and steroidogenesis in MA-10 mouse Leydig tumor cells. AB - Hormonal induction of steroidogenesis in the adrenal and gonads is dependent on the synthesis and function of the steroidogenic acute regulatory protein (StAR). As a first approach to investigate the role of translation in the control of StAR expression, we examined StAR protein synthesis and steroid production in MA-10 mouse Leydig tumor cells in the presence of the transcriptional inhibitor, actinomycin D. We show that human CG (hCG)-induced StAR synthesis, as determined by radiolabeling MA-10 cells with [35S]methionine and immunoprecipitation of StAR, is blocked by actinomycin D. The rate of hCG-stimulated progesterone production is also decreased, but not completely blocked, suggesting a possible StAR-independent mechanism that may contribute approximately 10-20% of the acute steroidogenic potential of the cells. When MA-10 cells were pretreated with hCG to increase StAR messenger RNA levels and then the proteins radiolabeled in the presence of hCG or hCG plus actinomycin D, no difference was observed in the amount of the 30-kDa StAR protein synthesized. However, a 50% increase in the precursor form of StAR protein was detected with hCG treatment alone. These data suggest that ongoing StAR protein synthesis is not inhibited by actinomycin D, but that continued synthesis requires transcriptional activity. Progesterone production was inhibited by actinomycin D in the hCG-pretreated cells, supporting the proposal that maintaining StAR protein synthesis is required for optimal steroid production in MA-10 mouse Leydig tumor cells. PMID- 9348221 TI - Coordinate expression of matrix metalloproteinase family members in the uterus of normal, matrilysin-deficient, and stromelysin-1-deficient mice. AB - The expression patterns of matrix metalloproteinase (MMP) family members during the murine estrous cycle and postpartum uterine involution were analyzed, and the consequence of removing specific MMPs during uterine functions was determined using mice deficient in either matrilysin (MAT) or stromelysin-1 (STR-1). In wild type animals, MAT, STR-1, STR-2, STR-3, and gelatinase A were consistently expressed during the most active phases of the estrous cycle, estrus and proestrus. The messenger RNA for these MMPs as well as collagenase-3 and the tissue inhibitors of metalloproteinases were also expressed during uterine involution, as determined by Northern analysis and in situ hybridization. Notably, MAT, STR-2, and collagenase-3 messenger RNA levels were elevated at early times of involution and rapidly decreased with time, whereas the transcripts for other MMPs remained elevated throughout the involution process. Involution proceeded normally in mice lacking MAT or STR-1; however, the expression of STR-1 and STR-2 was dramatically up-regulated in MAT nullizygous mice, and the expression of MAT and STR-2 was moderately up-regulated in STR-1 deficient animals. We conclude that the concerted action of several MMPs is likely to play an important role in the remodeling of the postpartum uterus, and that mechanisms that compensate for the loss of a specific MMP during this process appear to exist. PMID- 9348222 TI - Differential regulation of 11 beta-hydroxysteroid dehydrogenase type 1 and 2 by nitric oxide in cultured human placental trophoblast and chorionic cell preparation. AB - Two types of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) have been identified in different tissues. Type 1 has both oxidase and reductase activities interconverting cortisol and cortisone, whereas type 2 has only oxidase activity converting cortisol to cortisone. It has been proposed that placental 11 beta-HSD controls the passage of maternal glucocorticoids to the fetal circulation. However, little is known about the regulation of 11 beta-HSD in the human placenta and fetal membranes. We cultured human term placental trophoblast and chorionic trophoblast cells to examine effects of nitric oxide donors, sodium nitroprusside (SNP) and S-nitroso-N-acetyl penicillamine (SNAP), on the activity and messenger RNA (mRNA) expression of 11 beta-HSD. At 72 h of culture, placental trophoblast formed syncytial clumps that were cytokeratin positive and displayed mainly type 2 oxidase activity, although some type 1 reductase activity was detectable. Chorion preparations contain greater than 90% trophoblast cells as demonstrated by immunostaining for cytokeratin and less than 5% vimentin positive cells. Type 1 reductase activity predominated in the chorionic trophoblast cells with barely detectable type 1 or type 2 oxidase activity. Both SNP (1-400 microM) and SNAP (1 mM) inhibited placental 11 beta-HSD type 2 oxidase activity but not type 1 reductase activity either in placental or chorionic cells. An inhibitory effect on type 2 oxidase activity was reproduced in part by 8-bromo cGMP, blocked partially by the guanylate cyclase inhibitor LY83583 (1 microM), but not by an ADP-ribosylation inhibitor N, N'-hexamethylene-bis-acetamide (HMBG) (10 mM). SNP also suppressed the expression of type 2 mRNA in cultured placental trophoblast in a dose-dependent manner, and this effect was also blocked by LY83583. We conclude that human placental trophoblast possesses predominantly 11 beta-HSD type 2 oxidase activity, whereas chorionic cells possess mainly type 1 reductase activity under the culture conditions employed. Nitric oxide specifically attenuated 11 beta-HSD type 2 oxidase activity as well as its mRNA expression in the placental trophoblast. The effect was mediated at least partially through the cGMP pathway, although an alternative pathway other than ADP-ribosylation may exist. PMID- 9348223 TI - Regulation of the expression of the angiogenic enzyme platelet-derived endothelial cell growth factor/thymidine phosphorylase in endometrial isolates by ovarian steroids and cytokines. AB - The angiogenic enzyme platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) was strongly expressed in the endometrial glands in the luteal and menstrual, but not the proliferative, phases of the cycle. The converse was seen in the stroma, where expression was strong in the proliferative, but not the luteal or menstrual, phases. Inflammatory cytokines induced PD-ECGF/TP expression in primary cultures of human normal endometrial epithelial (NEE) and normal endometrial stromal cells. The profile of cytokine induction of PD-ECGF/TP was cell dependent. Thus, in NEE cells, PD-ECGF/TP expression was strongly induced by the combination tumor necrosis factor-a and interferon-gamma. In contrast, in normal endometrial stromal cells, interferon gamma gave, by far, the strongest induction of PD-ECGF/TP. Expression of the enzyme was not regulated by ovarian hormones alone. Although treatment of NEE cells with a physiological concentration of progesterone (5 X 10[-8] M) or transforming growth factor-beta1 (10 ng/ml) alone had no effect on PD-ECGF/TP expression, when delivered together at the same dose they induced a 48-fold increase in expression. This expression correlates with cyclic changes in progesterone and transforming growth factor-beta1 levels in the uterus. PMID- 9348224 TI - Cloning, tissue expression, and chromosomal localization of the mouse IRS-3 gene. AB - Insulin receptor substrate (IRS) proteins are key regulators of basic functions such as cellular growth and metabolism. They provide an interface between multiple receptors and a complex network of intracellular signaling molecules. Two members of this family (IRS-1 and IRS-2) have been identified previously. In this investigation, we analyzed a mouse expressed sequence tag clone that proved to be a new member of the IRS family. Sequence analysis of this clone and comparison with the sequences deposited in GenBank demonstrates this protein may be the murine homolog of rat IRS-3, recently purified and cloned from rat adipocytes. Accordingly, we have named our protein mouse IRS-3. The expressed sequence tag clone contains the complete coding sequence of 1485 bp, encoding a protein of 495 amino acids. Sequence alignment with the other members of the IRS family shows that this protein contains pleckstrin homology and phosphotyrosine binding domains that are highly conserved. In addition, there is conservation of many tyrosine phosphorylation motifs responsible for interactions with downstream signaling molecules containing SH2 domains. The murine IRS-3 messenger RNA (2.4 kilobases in length) is expressed in many tissues, with highest levels in liver and lung. Mouse IRS-3 is highly expressed in the first part of the embryonic life, when IRS-1 messenger RNA is barely detectable. Unlike the genes encoding IRS-1 and IRS-2, the IRS-3 gene contains an intron (344 bp in length) in the region between the pleckstrin homology and the phosphotyrosine-binding domains. Fluorescent in situ hybridization localized the mouse IRS-3 gene on the telomeric region of chromosome 5G2. Cloning of the murine IRS-3 gene will make it possible to apply genetic approaches to elucidate the physiological role of this new member of the IRS family of proteins. PMID- 9348225 TI - The small guanosine triphosphate-binding protein Rab4 is involved in insulin induced GLUT4 translocation and actin filament rearrangement in 3T3-L1 cells. AB - Insulin's stimulation of glucose transport involves the translocation of vesicles containing the glucose transporter GLUT4 to the plasma membrane. Small GTP binding proteins have been implicated in the regulation of vesicular traffic. We studied the effects of microinjection of wild-type Rab4 glutathione S-transferase fusion protein (WT Rab4), a GTP-binding defective mutant (Rab4 N121I), a guanosine triphosphatase-defective mutant (Rab4 Q67L), and a Rab4 antibody on insulin-induced GLUT4 translocation in 3T3-L1 adipocytes. Microinjection of Rab4 N121I and Rab4 antibodies had no effect on basal GLUT4 staining, but inhibited insulin-induced GLUT4 translocation by 50% compared with that in control IgG injected cells. WT Rab4 and Rab4 Q67L microinjection had no effect on either basal or insulin-induced GLUT4 translocation. Premixing and coinjection of the Rab4 antibody with WT Rab4 almost completely abolished its inhibitory effect on insulin-induced GLUT4 translocation. In contrast, microinjection of an antibody directed against the highly conserved region of Rab3 proteins had no effect on insulin-induced GLUT4. These results point to a direct role of Rab4 in insulin induced GLUT4 translocation, and that this effect is dependent on nucleotide binding to the protein. We also studied the effect of microinjection of the same proteins on insulin-induced actin filament rearrangement (membrane ruffling) in the same cell line. Microinjection of Rab4 N121I and Rab4 antibodies inhibited insulin-induced membrane ruffling by 40%, whereas WT Rab4 or a Rab3 antibody injection had no effect on cytoskeletal rearrangement. In summary, 1) Rab4 is a necessary component of the insulin/GLUT4 translocation signaling pathway; 2) the function of Rab4 in this pathway requires GTP binding; 3) Rab4 also participates in the process of insulin-induced membrane ruffling; and 4) Rab3 proteins do not seem to be involved in these processes. PMID- 9348226 TI - Evidence for functional roles of Crk-II in insulin and epidermal growth factor signaling in Rat-1 fibroblasts overexpressing insulin receptors. AB - We examined the potential role of Crk-II in insulin and epidermal growth factor (EGF) signaling in Rat-1 fibroblasts overexpressing insulin receptors. Crk is an SH2 and SH3 domain-containing adaptor protein that has been reported to associate with p130cas, paxillin, c-cbl, c-abl, Sos, and C3G in vitro. Insulin- and EGF induced association of Crk-II with these molecules was assessed by immunoblotting of anti-Crk-II precipitates in Rat-1 fibroblasts overexpressing insulin receptors. Neither insulin nor EGF treatment induced Crk-II association with either Sos or C3G. Basal tyrosine phosphorylation of c-abl and its constitutive association with Crk-II were not further increased by insulin or EGF. p130cas and paxillin were heavily tyrosine phosphorylated in the basal state. Both insulin and EGF stimulated their dephosphorylation, followed by p130cas-Crk-II dissociation and paxillin-Crk-II association, although the magnitude of these effects was greater with insulin than with EGF. Interestingly, EGF, but not insulin, stimulated tyrosine phosphorylation of c-cbl and its association with Crk-II. To investigate the functional roles of Crk-II in mitogenesis and cytoskeletal rearrangement, we performed microinjection analysis. Cellular microinjection of anti-Crk-II antibody inhibited EGF-induced, but not insulin induced, DNA synthesis. Insulin, but not EGF, stimulated cytoskeletal rearrangement in the cells, and microinjection of anti-Crk-II antibody effectively inhibited insulin-induced membrane ruffling, suggesting that Crk-II is involved in insulin-induced cytoskeletal rearrangement. These results indicate that Crk-II functions as a multifunctional adaptor molecule linking insulin and EGF receptors to their downstream signals. The presence of c-cbl-Crk-II association may partly determine the signal specificities initiated by insulin and EGF. PMID- 9348227 TI - Osteoclasts are present in gp130-deficient mice. AB - Interleukin (IL)-6, IL-11, leukemia inhibitory factor, and oncostatin M similarly induce osteoclast formation in cocultures of osteoblastic cells and bone marrow cells. These cytokines share a common signal transducer, gp130, which forms a receptor complex with the specific receptor for each cytokine. To investigate the role of gp130 in osteoclast development, we examined bone tissues in gp130 deficient and wild-type newborn mice of the ICR background. Soft x-ray radiographs and microfocus x-ray computed tomographs revealed that bone marrow cavities were present in tibiae and radii of both wild-type and gp130-deficient mice. Microfocus x-ray computed tomography and histological examination demonstrated a decrease in the amount of trabeculae at the metaphysial region in tibiae and radii of the gp130-deficient mice compared with the wild-type mice. The number ofosteoclasts in gp130-deficient mice was about double that in the wild-type mice. There were no apparent differences in the distributions of alkaline phosphatase-positive osteoblasts and the osteoid surface on the trabecular bone at the metaphysial region between the wild-type and gp130 deficient mice. The volume of mineralized trabecular bones was also decreased at mandibulae, accompanied by the increased number of osteoclasts in gp130-deficient mice compared with the wild-type and heterozygous mice. These results suggest that the formation of osteoclasts is not solely dependent on gp130 signaling, at least during fetal development. The osteoclastic bone resorption in gp130 deficient mice may be caused by the functional redundancy of bone-resorbing hormones and cytokines other than those of the IL-6 family. PMID- 9348228 TI - Modulation of commitment, proliferation, and differentiation of chondrogenic cells in defined culture medium. AB - The factors regulating the growth and development of mesenchymal precursor cells toward chondrogenesis are not well identified. We have developed a defined serum free culture system that allows the proliferation of chick embryo chondrogenic cells and their maturation toward hypertrophic chondrocytes. Proliferation is obtained in adhesion in medium supplemented with insulin (Ins), Dexamethasone (Dex), and either basic fibroblast growth factor (FGF-2), platelet-derived growth factor bb, epithelial growth factor, or GH; the highest mitogenic response is induced by FGF-2 in synergy with Ins. Ins can be substituted by Ins-like growth factor I. When these cells are transferred into suspension culture in Ins/Dex and T3 without growth factor supplement, they undergo the complete chondrogenic development characterized by type X collagen synthesis and cellular hypertrophy. During differentiation, Ins cannot be substituted by Ins-like growth factor I. Chondrogenesis is also evidenced by the formation of hypertrophic cartilage when the medium is supplemented with ascorbic acid. If T3 is introduced in the proliferation phase, the cells fail to differentiate to hypertrophy in suspension unless bone morphogenetic protein-2 is added. Assays of ectopic tissue formation in nude mice, with cells implanted sc after adsorption on collagen sponge or porous hydroxyapatite ceramics, indicate that cells grown in Ins/FGF-2 reform mainly cartilage in vivo, whereas expansion in Ins/T3/Dex/FGF-2 leads to the formation of cartilage, bone, and adipose tissue. PMID- 9348229 TI - Endoglin regulates trophoblast differentiation along the invasive pathway in human placental villous explants. AB - Successful invasion of the maternal vascular system by trophoblast cells is a prerequisite for the establishment of a normal hemochorial placenta. Transforming growth factor-beta (TGFbeta) has been implicated in the regulation of trophoblast invasiveness into the uterus. Endoglin is a component of the TGFbeta receptor complex that binds beta1 and beta3 isoforms and is expressed at high levels on syncytiotrophoblast throughout pregnancy and is also transiently up-regulated on extravillous trophoblasts differentiating along the invasive pathway. We investigated the role of endoglin in a serum-free human villous explant culture system that allows the study of trophoblast outgrowth, migration, and invasion and mimics events occurring in anchoring villi during the first trimester of gestation. Addition to explant cultures from 5-8 weeks gestation of a monoclonal antibody to endoglin or of antisense endoglin oligonucleotides significantly stimulated trophoblast outgrowth and migration. These responses were specific, as incubation of explants with nonimmune IgG or sense and scrambled oligonucleotides had no effect. Antisense endoglin-induced trophoblast outgrowth and migration were accompanied by cell division of villous-associated trophoblasts within the proximal region of the forming column and by the characteristic switch in integrins observed in anchoring villi in situ. Treatment of villous explants with antibody and antisense oligonucleotides to endoglin also resulted in an increased fibronectin release into the culture medium. The stimulatory effect of antisense endoglin on fibronectin production was overcome by the addition of exogenous TGFbeta2, but not TGFbeta1 and -beta3. These findings suggest that endoglin expression in the transition from polarized to nonpolarized trophoblasts in anchoring villi is necessary for mediation of the inhibitory effect of TGFbeta1 and/or TGFbeta3 on trophoblast differentiation along the invasive pathway. PMID- 9348230 TI - Sorbin in the porcine gastrointestinal tract and pancreas: an immunocytochemical analysis. AB - Sorbin is a 153-amino acid peptide that was initially discovered in the porcine duodenum. We have reported previously that this peptide regulates intestinal electrolyte transport and have described accumulation sites in the rat digestive tract. In the present study, we investigated the anatomical distribution and the site(s) of sorbin production in the porcine digestive tract using immunocytochemistry. The use of polyclonal antisera, which by cross-reaction studies were shown to be specific for different regions of the molecule, revealed a diversified distribution. Sorbin predominated in endocrine cells preferentially localized in the pyloric glands, duodenal crypts of Lieberkuhn, and pancreatic islets; in the gastrointestinal tract, sorbin coexisted with Met-enkephalin or with substance P in a small fraction of serotonin-storing [enterochromaffin (ED)] cells, i.e. EC2 cells and EC1 cells, respectively; in the pancreas, sorbin coexisted with insulin in the beta-cells, also considered as serotonin-storing cells in the pig, and with EC cells in the exocrine pancreas. An enteric neuronal system containing sorbin was also reported. Our results demonstrate that sorbin is a component of the serotonin-storing cell type in the porcine gastrointestinal tract and pancreas, and suggest potential directions to investigate the functions of this new regulatory peptide. PMID- 9348231 TI - Role of androgens in testicular tumor development in inhibin-deficient mice. AB - To understand gonadal tumor development, we have previously created a mouse model in which mice deficient in the inhibins develop gonadal sex cord-stromal tumors with essentially 100% penetrance. These tumors develop as early as 4 weeks of age and cause cancer cachexia-like symptoms and subsequent death in the inhibin deficient mice. Gonadectomized inhibin-deficient mice eventually develop adrenal cortical tumors with nearly 100% penetrance. These studies have identified inhibin as a novel secreted tumor suppressor protein with specificity for the gonads and adrenal glands. Sex steroids have been implicated to influence gonadal tumor development in humans and mice. To determine the role of androgens in gonadal tumorigenesis in inhibin-deficient male mice, we have used a genetic intercross strategy, breeding inhibin alpha mutant mice with tfm (testicular feminization, a naturally occurring androgen receptor mutant) carrying females to eventually generate compound mutant male mice that lack inhibins and carry the tfm mutation. These compound mutant mice, like inhibin-deficient mice, continue to develop testicular tumors and the accompanying cancer cachexia-like wasting syndrome. Consistent with these findings, elevated levels of activins A and B secreted from the gonadal tumors are seen in the adult compound mutant mice as well as the secondary pathological consequences of these high activin levels in the livers and glandular stomachs. However, in contrast to male mice lacking only inhibin, in which essentially 100% of the testicular tumors are hemorrhagic, 65% of the tumors in these compound mutant male mice are less hemorrhagic, and approximately 50% of the compound mutants live longer than 17 weeks of age (95% of the male mice lacking only inhibin die by 12 weeks). These results suggest that androgens are not required for testicular tumor development in inhibin deficient mice, but may play a regulatory role in testicular tumor progression. PMID- 9348232 TI - Cellular localization and regulation of expression of testicular estrogen sulfotransferase. AB - Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes the specific sulfonation of estrogens at the 3-hydroxyl position using 3'-phosphoadenosine-5' phosphosulfate as an activated sulfate donor. Sulfated estrogens no longer bind to the estrogen receptor and are, therefore, hormonally inactive. Although liver has been considered a primary site for steroid sulfotransferase activities, we previously have cloned the mouse EST complementary DNA and found the enzyme to be expressed abundantly in the testis of normal mice. In this study we show by reverse transcription-PCR that EST is also expressed in the testes of rat and man, suggesting that testicular expression of EST may be a common phenomenon among different species. Using a purified polyclonal antibody raised against the bacterially expressed mouse EST protein, we demonstrate by immunohistochemistry that EST is localized selectively to the androgen-producing Leydig cells within the mouse testis. Additionally, we show that Leydig cell expression of EST is under the control of the pituitary hormone LH and is regulated differentially during development. In contrast to the high level of expression in mature intact animals, EST is not present in Leydig cells of hypophysectomized mice or in Leydig cells of fetal and prepubertal (day 5 or 17) mouse testes. Administration of hCG to hypophysectomized mice restored the testicular expression of EST. Together, these results suggest that testicular expression of EST may play an important role in male reproduction, conceivably by modulating the activity of locally synthesized estrogen in the testis of a sexually mature animal. PMID- 9348233 TI - Immunocytochemical localization of the NPY/PYY Y1 receptor in the developing pancreas. AB - Neuropeptide Y, peptide YY, and pancreatic polypeptide are structurally related peptides that are considered to play a role in the regulation of pancreatic secretion and blood flow. Several receptor subtypes for these peptides have been identified, and the Y1, Y2, Y4/PP1, Y5, and Y5/PP2/Y2b receptors are cloned. We have prepared polyclonal peptide antibodies that recognize the Y1 receptor and now report on its localization in the adult and developing rat pancreas. In the adult pancreas, Y1 receptors were detected both in some centroacinar and intralobular duct cells and in endothelial cells. In the developing pancreas (E12.5-E16.5), Y1 receptor immunoreactivity was observed in numerous nonendocrine epithelial cells. These cells occurred in the immediate vicinity of peptide YY positive endocrine cells. At E16.5, a fraction of these Y1 receptor-containing cells co-stored amylase. One day later, Y1 receptor immunoreactivity became restricted to pancreatic duct-like cells that occurred in close proximity to peptide YY cells. In fetal rats, intense Y1 receptor staining was also observed in endothelial cells. These observations, together with the finding of early pancreatic peptide YY expression, suggest that peptide YY produced by fetal endocrine cells may exert an action on exocrine cells, duct cells and endothelial cells during development. PMID- 9348235 TI - Expression and subcellular distribution of the beta-isoform of the human glucocorticoid receptor. AB - Alternative splicing of the human glucocorticoid receptor (hGR) primary transcript produces two highly homologous protein isoforms, termed hGR alpha and hGRbeta, that differ at their carboxy-termini. In contrast to the well characterized hGR alpha isoform, which modulates gene expression in a hormone dependent fashion, the biological significance of hGRbeta has only recently begun to emerge. We and others have shown that the hGRbeta messenger RNA transcript is widely expressed in human tissues and that the hGRbeta protein functions as a dominant negative inhibitor of hGR alpha in transfected cells. Unfortunately, these initial studies did not determine whether the hGRbeta protein was made in vivo. Such analyses are hindered because available anti-hGR antibodies cannot discriminate between the similarly sized hGR alpha and hGRbeta proteins. Therefore, to investigate the expression of the hGRbeta protein, we have produced an antipeptide, hGRbeta-specific antibody termed BShGR. This antibody was made against the unique 15-amino acid peptide at the carboxy-terminus of hGRbeta and recognizes both the native and denatured conformations of hGRbeta, but does not cross-react with hGR alpha. Using BShGR on Western blots and in immunoprecipitation experiments, we detected the hGRbeta protein in a variety of human cell lines and tissues. Immunocytochemistry was then performed with BShGR on HeLa S3 and CEM-C7 cells and on tissue sections prepared from lung, thymus, and liver to assess the cellular and subcellular distribution of hGRbeta. In all immunopositive cells, hGRbeta was found in the nucleus independent of glucocorticoid treatment. Within tissues, the hGRbeta protein was expressed most abundantly in the epithelial cells lining the terminal bronchiole of the lung, forming the outer layer of Hassall's corpuscle in the thymus, and lining the bile duct in the liver. As a potential in vivo inhibitor of hGR alpha activity, expression of hGRbeta may be an important factor regulating target cell responsiveness to glucocorticoids. PMID- 9348234 TI - Fas (APO-1, CD95)-mediated apoptosis in thyroid cells is regulated by a labile protein inhibitor. AB - To determine whether thyroid cell apoptosis observed in autoimmune thyroid disease could be related to activation of the Fas pathway, we examined the expression and function of Fas on thyroid follicular cells in vitro. Fas messenger RNA was found to be present using two different techniques and was expressed at equal levels in thyrocytes cultured either in the presence or absence of TSH. Fas antigen protein expression was demonstrated by Western blot of thyroid cell lysates and by immunohistochemical staining of thyrocytes, and the amount of Fas protein present did not appear to vary regardless of culture conditions. Despite expressing substantial amounts of Fas protein, thyrocytes treated with anti-Fas monoclonal antibody failed to undergo apoptosis. The addition of either interferon-gamma or interleukin-1beta to the anti-Fas-treated cell cultures also did not promote apoptotic signaling through this pathway. In contrast, the concomitant administration of cycloheximide allowed the induction of apoptosis through the activation of Fas in thyrocytes. These results suggest that Fas is constitutively expressed in thyrocytes, but that the induction of apoptosis through the Fas pathway is blocked by a labile protein inhibitor. PMID- 9348236 TI - Thyroid hormone receptor (alpha) distribution in hamster and sheep brain: colocalization in gonadotropin-releasing hormone and other identified neurons. AB - Thyroid hormones appear to play an important role in the seasonal reproductive transitions of a number of mammalian and avian species. These seasonal transitions as well as the effects of thyroid hormones on the reproductive neuroendocrine axis are mediated by the GnRH system. How thyroid hormones affect the GnRH system is unclear. Double label immunocytochemistry was used to examine GnRH- and other neurotransmitter/neuropeptide-containing neurons for thyroid hormone receptor (alphaTHR) colocalization in two seasonal breeders, the golden hamster and the sheep. AlphaTHR was identified in hamster and sheep brain by Western blot analysis. Furthermore, alphaTHR immunoreactivity was widely distributed in brain and was colocalized in identified populations: GnRH neurons (hamster, 28%; sheep, 46%); dopaminergic neurons of the A14 (hypothalamic) and A16 (olfactory bulb) cell groups, but not in the hypothalamic A13 cell group; and neurophysin-immunoreactive neurons of the supraoptic and paraventricular nuclei. The finding of alphaTHR in GnRH and A14 dopamine neurons provides an anatomical substrate for direct thyroid hormone action on the reproductive neuroendocrine system of these two seasonally breeding species. It remains to be determined whether the GnRH gene itself or the gene of another constituent within the same GnRH neuron is responsive to thyroid hormones. PMID- 9348238 TI - Transcription factor Sp1 is necessary for basal calmodulin gene transcription and for its selective stimulation by insulin. AB - Insulin positively regulates transcription of rat calmodulin (CaM) I gene and activates the low Km cyclic AMP (cAMP) phosphodiesterase (PDE). To elucidate the mechanism of transcriptional regulation, rat hepatoma (H-411E) cells were transfected with DNA constructs containing the putative CaM promoters coupled to a luciferase reporter and challenged with insulin. Activation of the full length 1835 bp rat CaM I promoter containing all three Sp1 sites or truncated promoters with combinations of one to three of the Sp1 sites was studied in Sp1 deficient Drosophilia SL2 cells and in SL2 cells co-transfected with an Sp1 expression vector and re-challenged with insulin. Our results demonstrate that Sp1 is obligatory for basal activation of the CaM promoter. The maximal insulin stimulation of CaM promoter is elicited only if it contains at least two Sp1 sites. PMID- 9348237 TI - Corticotropin releasing factor receptor type II (CRF2) messenger ribonucleic acid levels in the hypothalamic ventromedial nucleus of the infant rat are reduced by maternal deprivation. AB - The stress neurohormone corticotropin releasing factor (CRF) activates at least two receptor types. Expression of corticotropin releasing factor receptor type II (CRF2) has been demonstrated in the hypothalamic ventromedial nucleus (VMH) of the adult and developing rat, but the physiological functions of VMH-CRF2 have not been elucidated. The VMH has been documented as an important participant in the regulation of food intake and its interactions with the hypothalamic pituitary-adrenal axis and circadian rhythms. Regulation of VMH-CRF2 may thus play a role in the interplay of physiological alterations in metabolic state with the neuroendocrine and anorexic effects of CRF. This study determined the regulation of CRF2-mRNA expression in infant rats by the physiological consequences of maternal deprivation, i.e., fasting and stress. Using in situ hybridization, maternally deprived pups had an average 62% reduction of VMH-CRF2 mRNA levels compared with stress-free controls. Maternal deprivation also resulted in elevated plasma corticosterone levels (3.8 +/- 0.3 vs. 1.3 +/- 0.1 microg/dl) and an average 5.7% body weight loss. This study demonstrates that maternal deprivation, via fasting and HPA activation, leads to a dramatic decrease of CRF2-mRNA levels in the VMH. These results are consistent with a role for CRF2 activation in mediating some of the complex interactions of CRF (or urocortin) with regulation of food intake in the developing rat. PMID- 9348239 TI - Nitric oxide mediates leptin-induced luteinizing hormone-releasing hormone (LHRH) and LHRH and leptin-induced LH release from the pituitary gland. AB - Previous experiments have demonstrated that leptin releases luteinizing hormone releasing hormone (LHRH) from median eminence (ME)-arcuate explants from male rats and also stimulates the release of follicle-stimulating hormone (FSH) and LH from anterior pituitaries with a potency not significantly different from that of LHRH itself. To determine the mechanism by which leptin acts at both the hypothalamic and pituitary level, we evaluated the effect of a competitive inhibitor of nitric oxide synthase (NOS), NG-monomethyl-L-arginine (NMMA) on the response to leptin. To evaluate the role of NO in the action of leptin to release LHRH, ME-arc explants were incubated with leptin (10[-11] M), a concentration shown earlier to give the most effective stimulation of LHRH release. NMMA (3 x 10[-4] M) completely inhibited the LHRH release induced by leptin. In other experiments, hemi-anterior pituitaries were incubated with NMMA with and without leptin at various concentrations (10[-9] - 10[-6] M). As in the case of hypothalamic explants, NMMA had no effect on basal release of LH; however, it completely blocked the stimulation of LH release induced by leptin. Interestingly, the release of LH induced by LHRH (4 x 10[-9] M) was also completely blocked by the inhibitor of NOS. The results provide evidence that leptin acts both at hypothalamic and pituitary level to stimulate NO release, presumably by acting on its receptors at both sites which then induces the release of either LHRH or LH, respectively. Furthermore, LH release induced by LHRH is also mediated by NO. PMID- 9348240 TI - An autocrine progesterone positive feedback loop mediates oxytocin upregulation in bovine granulosa cells during luteinization. AB - During luteinization of bovine granulosa cells in vitro in the presence of insulin, or insulin plus forskolin, there is a massive upregulation not only of progesterone production, but also of the gene for the peptide hormone oxytocin, with secretion of the peptide into the medium. By using the progesterone receptor antagonists, RU486 or onapristone, we have shown that this upregulation of the oxytocin gene is mediated by an autocrine loop involving endogenous progesterone and the progesterone receptor in the granulosa cells. The inhibition of oxytocin gene expression by the anti-gestagens can be reversed by medroxyprogesterone acetate but not by dexamethasone, showing that the effect is due to the endogenous progesterone. Since oxytocin also appears to be involved in a positive feedback loop regulating steroid output, these results show that endogenous progesterone itself is a key feedback element in the cascade of events termed luteinization, and that possibly anti-gestagens can influence ovarian function in vivo by directly disrupting this process. PMID- 9348242 TI - Differential expression of estrogen receptors alpha and beta mRNA during differentiation of human osteoblast SV-HFO cells. AB - Estrogens have been shown to be essential for maintaining a sufficiently high bone mineral density and ER alpha expression has been demonstrated in bone cells. Recently, a novel estrogen receptor, estrogen receptor beta (ERbeta) has been identified. Here we demonstrate that also ERbeta is expressed in human osteoblasts, and that ER alpha and ERbeta are differentially expressed during human osteoblast differentiation. ERbeta mRNA expression increased gradually during osteoblast culture, resulting in an average increase of 9.9+/-5.3 fold (mean+/-S.D., n=3) at day 21 (mineralization phase) as compared to day 6 (proliferation phase). In contrast, ER alpha mRNA expression levels increased only slightly until day 10 (2.3+/-1.7 fold) and then remained constant. The observed differential regulation of ER alpha and beta is suggestive for an additional functional role of ERbeta to ER alpha in bone metabolism. PMID- 9348241 TI - Regulation of hypothalamic proopiomelanocortin mRNA by leptin in ob/ob mice. AB - The hormone leptin acts on the brain to regulate feeding, metabolism, and reproduction; however, its cellular targets and molecular mechanisms of action remain to be fully elucidated. The melanocortins, which are derived from the precursor proopiomelanocortin (POMC), are also implicated in the physiological regulation of body weight. POMC-containing neurons express the leptin receptor, and thus it is conceivable that the POMC gene itself may be part of the signaling pathway involved in leptin's action on the brain. Using in situ hybridization and computerized image analysis, we tested the hypothesis that the POMC gene is a target for regulation by leptin by comparing cellular levels of POMC mRNA in the hypothalamus among groups of leptin-deficient (ob/ob) mice, leptin-treated ob/ob mice, and wild-type controls. POMC mRNA levels were significantly reduced throughout the arcuate nucleus in vehicle-treated ob/ob mice relative to wild type controls, whereas POMC mRNA levels in leptin-treated ob/ob mice were indistinguishable from wild-type controls. These observations suggest that one or more products of POMC serve as an integrative link between leptin and the central mechanisms governing body weight regulation and reproduction. PMID- 9348244 TI - Quantitative analysis of growth hormone (GH) pre-mRNA expression in cultured rat anterior pituitary cells by an intron-specific and competitive PCR method. AB - We have developed a novel method of quantifying growth hormone(GH) pre-mRNA expression in anterior pituitary cells. DNA-free total RNA extracted from cultured rat anterior pituitary cells was reverse transcribed(RT) to cDNA, and RT products were subsequently quantitated by competitive PCR using intron-specific primers of rat GH gene. After 6-h of incubation in treated cells, dexamethasone(Dex) and triiodo-L-thyronine(T3) significantly increased GH pre mRNA levels(3.2- and 2.2-fold compared to non-treated cells, respectively). However, Northern blot analysis did not detect significant changes in GH mRNA levels. After 24-h incubation with Dex and T3, significant increases in GH mRNA levels were detected on Northern blots, but GH pre-mRNA levels did not differ between treated and non-treated cells. These findings suggest that both Dex and T3 treatments rapidly increase GH pre-mRNA levels in normal somatotropes. This method has high sensitivity and widespread application to the analysis of pre mRNAs of target genes. PMID- 9348243 TI - Autocrine effect of androgen on proliferation of an androgen responsive prostatic carcinoma cell line, LNCAP: role of interleukin-6. AB - LNCaP is an androgen-responsive prostatic carcinoma cell line that exhibits a bell-shaped growth response curve to increasing doses of dihydrotestosterone (DHT) in culture. Although the precise mechanism responsible for this unique growth response to androgen stimulation remains unclear, many studies have suggested that androgen modulates the level of various growth factors. In an early study, we demonstrated that LNCaP proliferation was stimulated by interleukin (IL)-6 in a paracrine manner, because these cells did not express a significant amount of IL-6. In the present study, the role of IL-6 in mediating androgen regulated proliferation in LNCaP cells was investigated. DHT, at increasing doses up to 10(-8) M, resulted in a release of IL-6 from LNCaP cells. This dose-dependent effect of DHT on LNCaP proliferation could be partially inhibited by the addition of antibody against IL-6 into the culture medium. These results indicate that the DHT-induced expression of IL-6 stimulates proliferation of LNCaP cells in culture in an autocrine manner. PMID- 9348245 TI - Transient ubiquitin cross-reactive protein gene expression in the bovine endometrium. AB - Bovine ubiquitin cross-reactive protein (boUCRP) is secreted by the endometrium from days 15 to 26 of pregnancy in response to conceptus-derived interferon-tau (IFN-tau). We hypothesized that the gene encoding boUCRP was under transcriptional control by the conceptus and IFN-tau. Northern blots using radiolabeled UCRP cDNA revealed a single UCRP transcript of approximately 700 b that was present (P < 0.05) in endometrial cells cultured with 25 nM rboIFN-tau. The UCRP mRNA was not detected in endometrium on days 15, 17, 18 or 19 of the estrous cycle (n = 4 cows on each day) or in spleen, kidney, liver, corpus luteum or muscle. Bovine UCRP mRNA was detectable (P < 0.05) in endometrium from pregnant cows by day 15, reached highest levels by day 17, remained elevated on days 18, 19 and 21, and then declined to amounts on day 26 that were not detectable. Northern blot using radiolabeled ubiquitin cDNA revealed presence of the two major ubiquitin transcripts UbB (1.2 Kb) and UbC (2.6 Kb) in all tissues examined. The bovine UCRP cDNA did not cross-hybridize with these ubiquitin transcripts. We conclude that transcription of the UCRP gene is transient during early pregnancy and regulated by IFN-tau. PMID- 9348246 TI - Internet and obstetrics and gynecology. AB - BACKGROUND: The Internet is an unrestricted, easily accessed source of information that has been greatly touted by the mass media. It represents a major advance in information acquisition and dissemination. It is estimated that between 3040 million North Americans alone have access to the Internet. This multimedia reservoir of information has developed into a significant resource for information gathering, especially for those who depend on comprehensive and contemporary information, including scientists and physicians. METHODS AND RESULTS: The purpose of this article is to describe some of the basic and practical services available on the Internet including e-mail, literature searches, interest groups, databases and on-line journals. Our focus is to describe these Internet resources with the physician in mind, and with particular emphasis on Internet applications for the obstetrician/gynecologist. CONCLUSION: The role of the Internet in medicine has not been defined, but continues to evolve. In the field of obstetrics and gynecology, the Internet has already become incorporated in various ways and it is safe to say that we are currently going through a revolution as fundamental to the human civilization as the eighteenth century industrial revolution--the information revolution. PMID- 9348247 TI - Immunohistochemical localization of nitric oxide synthase in human fetal membranes. AB - SUBJECT: It has been demonstrated that L-arginine-nitric oxide (NO) system is present in the myometrium during pregnancy where it can regulate uterine contractility. MATERIAL AND METHOD: We have studied by immunohistochemistry the localization of constitutive endothelial nitric oxide synthase (ecNOS) and brain nitric oxide synthase (bNOS) in human fetal membranes in term non laboring women. RESULTS: The amniotic epithelium and the trophoblast of chorion layer stained intensively for bNOS, while for ecNOS the immunoreactivity was weak and restricted to trophoblast cells. CONCLUSION: These findings are suggestive for NO production by the fetal membranes in term pregnancy. PMID- 9348248 TI - Transplacental needle passage and other risk-factors associated with second trimester amniocentesis. AB - OBJECTIVE: To assess the obstetric outcome of all pregnancies undergoing midtrimester amniocentesis over a 10 year period at one center and the risk factors for pregnancy loss associated to the procedure. MATERIAL AND METHOD: All 2083 pregnancies with known pregnancy outcome and second trimester amniocentesis were included. Risk-factors for pregnancy loss were analysed by using patients' charts and a special record from the amniocentesis. RESULTS: The over-all risk of pregnancy loss after second trimester amniocentesis was 1.3% (28/2083). There was a slight but nonsignificant relationship between the degree of experience of the gynecologist and risk for pregnancy loss. A more experienced operator used significantly fewer needle insertions (p<0.001). Multiple needle insertions were also associated with a slight, albeit nonsignificant, increase in incidence in fetal loss (3.8% after three or more insertions vs. 1.2% after one insertion, NS). No difference in spontaneous abortion incidence was found in patients having an anterior versus a posterior placenta, nor did transplacental needle passage increase the risk for pregnancy loss. Comparison between use of real-time ultrasonic guidance at the amniocentesis and static ultrasonography immediately prior to the procedure did not reveal any differences in the incidence in spontaneous abortion. CONCLUSION: Second trimester amniocentesis seems to be a safe method for prenatal diagnosis. The risk for pregnancy loss was low (1.3%) and was only slightly and nonsignificantly affected by the operator's experience and multiple needle insertions. Transplacental needle passage did not affect the risk of pregnancy loss. PMID- 9348249 TI - Concentration of anti-D antibodies in Rh(D) alloimmunized pregnant women, as a predictor of anemia and/or hyperbilirubinemia in their newborn infants. AB - OBJECTIVES: To define a simple, safe and reliable program for the monitoring of anti-D alloimmunized pregnancies by analysis of the covariation between antenatal values of the titer and the concentration of anti-D antibodies in maternal serum, the deltaOD(450 nm) in amniotic fluid samples, and the levels of B-hemoglobin and S-bilirubin in the newborns at birth. SUBJECTS: Ninety-three Rh(D) negative women with anti-D antibody titers > or = 16 who, after the completed 34th gestational week, gave birth to Rh(D) positive babies with a positive direct antiglobulin test. METHODS: The titers and the concentrations of anti-D antibodies in maternal serum were determined by standard procedures every second week from the 25th week of gestation. In 47 of the 93 women, deltaOD(450 nm) in amniotic fluid was determined at least once. All antenatal values used in the study were determined within 14 days before delivery. RESULTS AND CONCLUSION: Maternal serum antibody titers < or = 32 or > or = 1000 could in themselves well predict unaffected and affected newborns. Antibody titers between 64 and 512 could not accurately predict newborns with or without hemolytic disease. As a complementary monitoring test, determination of the deltaOD(450 nm) was found to be less accurate when compared to determination of the concentration of anti-D antibodies. In order to monitor Rh(D) alloimmunized pregnancies, determination of the concentration of anti-D antibodies may possibly replace determination of deltaOD(450 nm). PMID- 9348250 TI - Is a speculum examination sufficient for excluding the diagnosis of ruptured fetal membranes? AB - OBJECTIVE: To determine the false negative rate of a sterile speculum examination for the diagnosis of rupture of the membranes in women not in labor and without visible amniotic fluid at speculum examination. Furthermore, possible risks to the mother and the baby after suspected rupture of the membranes were analyzed. STUDY DESIGN: In women not in labor with suspected rupture of the membranes between gestational weeks 34 and 42, a sterile speculum examination was performed. If no amniotic fluid was visible, a test for Diamine oxidase was carried out. The results of tests were not known to the obstetricians or the women. The women were allowed to return home with no further controls if no amniotic fluid was visible at the speculum examination. Neonatal and obstetric outcome was recorded prospectively. RESULTS: Of 27,502 deliveries, 2,099 women not in labor attended the delivery ward for suspected rupture of the membranes after week 34. Amniotic fluid was visualized in 1,580 women. In 519 women in whom no amniotic fluid was seen at the speculum examination, the Diamine oxidase test was negative in 456 and positive in 63. Antibiotics were given to eleven children (2.4%) in the group with a negative Diamine oxidase and to one infant (1.6%) in the positive Diamine oxidase group (p>0.05). No differences in obstetric outcome were recorded. CONCLUSIONS: The false negative rate of a speculum examination for the diagnosis of rupture of the membranes in women without amniotic fluid visible at a speculum examination was 12% when Diamine oxidase was used as the standard for the diagnosis of rupture of the membranes. This study did not show any disadvantages for mothers and infants if the women were sent home after a false negative speculum examination. The value of biochemical methods in the management of women not in labor with rupture of the membranes after thirty-four weeks of gestation could be questioned. PMID- 9348251 TI - Preterm premature rupture of membranes and the associated risk for placental abruption. Inverse correlation to gestational length. AB - OBJECTIVE: To evaluate the association between prolonged premature rupture of membranes (PROM) and placental abruption, especially during midtrimester pregnancy. METHODS: A retrospective hospital based study of 83 women with PROM occurring between 14 and 32 weeks of gestation and where active expectant management was undertaken. RESULTS: Increased frequency of placental abruption was found in patients with early rupture of membranes. The incidence was 50% and 44% when rupture of the membranes occurred before 20 weeks or between 20-24 weeks of pregnancy, respectively. When PROM occurred during gestational ages of 29-32 weeks, the incidence was 13%. Patients with antepartum bleeding, both before (relative risk 34) and after rupture of the membranes (relative risk 38) had a significantly higher risk for placental abruption (p<.001) than women without bleeding prior to delivery. The overall neonatal survival rate was 87%. No neonatal deaths were considered to be directly caused by asphyxia due to placental abruption. CONCLUSION: Using active expectant management and strict routines it seems possible to minimize the risk for perinatal asphyxia and mortality. The clinician should be aware of the significant association between preterm premature rupture of membranes and the risk for subsequent placental abruption, especially in patients with early midtrimester PROM and history of bleedings before rupture of membranes or bleedings during the latency period. PMID- 9348252 TI - Sick leave and social benefits during pregnancy--a Swedish-Norwegian comparison. AB - BACKGROUND: To analyze the correlation between sickness absence, working conditions, pregnancy outcomes and pregnancy associated social benefits in two urban pregnant populations in Sweden and Norway with different social benefit systems. METHODS: Relevant information on 1649 delivered women was manually extracted by the authors from the antenatal care and delivery records as well as from the personal social security files kept in the Varnamo and Hamar communities, and then computerized in a depersonalized form. RESULTS: The reproductive histories and the pregnancy outcomes appeared clinically similar in the two samples. Swedish pregnant women were significantly more often employed outside home (84 vs. 69 per cent). The types of occupations held were similar in Varnamo and Hamar. Swedish pregnant women were significantly more sick-listed during pregnancy than Norwegian women (64 vs. 32 per cent) and with a longer average duration of the sick-leave spells (61 days vs. 44 days). The sick-leave rate among Swedish employed pregnant women was 75 per cent as compared to 48 per cent in Norway. The differences appeared most evident in younger pregnant women (<25 years). The Swedish sick-leave rates were higher within all four occupational subgroups studied. During the observation period the pregnancy associated social benefits were significantly more generous in Sweden. CONCLUSIONS: Sickness absence during pregnancy does not seem to covariate in a simple way with ill health, working conditions or the amount of social benefits available. The increased sick-leave rates in Sweden may possibly be accounted for by a changing attitude towards pregnancy and its natural consequences, especially among younger women. PMID- 9348253 TI - The clinical outcome in pregnancies of grand grand multiparous women. AB - OBJECTIVE: To longitudinally evaluate maternal and neonatal complications with relation to birth order, with specific emphasis on grand grand multiparity (at least 10th para). METHODS: The maternal and neonatal outcome of 1200 pregnancies/deliveries in 96 grand grand multiparas was longitudinally investigated in 4 stages of the mothers' life: the primiparas, the multiparas (2nd-5th paras), the grand multiparas (6th-9th paras) and the grand grand multiparas stage. RESULTS: The frequency of hypertension, diabetes, placental complications, operative interventions at delivery, macrosomic infants, chromosomal abbreviations and fetal/neonatal anomalies increased with increasing birth order, being at a maximum in grand grand multiparas. The preterm delivery and perinatal mortality rate did not differ between the 3 groups of multiparas. Perinatal outcome was good in each group. CONCLUSIONS: Grand grand multiparity carries the risk of hypertensive and diabetic complications, which, in turn, often lead to induced or operative deliveries and placental complications. However, grand grand multiparity is not a major problem in societies with a good maternal health care system. PMID- 9348254 TI - Symptom-giving pelvic girdle relaxation of pregnancy, postnatal pelvic joint syndrome and developmental dysplasia of the hip. The Norwegian Association for Women with Pelvic Girdle Relaxation (Landforeningen for Kvinner Med Bekkenlosningsplager). AB - OBJECTIVES: To describe the clinical characteristics and outcomes of a large group of women with symptom-giving pelvic girdle relaxation of pregnancy and postnatal pelvic joint syndrome. To determine if there is an increased incidence of developmental dysplasia of the hip in the children of women with such pelvic problems. METHODS: A postal survey of 1,609 Norwegian women registered as having pregnancy-initiated pelvic joint pain. The response rate was 79% and from the answers 1,115 women were defined as having had symptom-giving pelvic joint syndrome of pregnancy and/or postnatal pelvic joint syndrome. RESULTS: Pelvic pains began in the first pregnancy in 74% of the respondents usually beginning in the first trimester. Pelvic pain worsened with subsequent pregnancies and persisted for a mean of 6.25 years, often causing major incapacity and lifestyle changes. Rest and physical supports brought temporary relief only. Sacroiliac joints and the symphysis pubis were the commonest sites of pain but peripheral joints were also often affected. There was a strong family history of both pelvic joint syndrome and developmental dysplasia of the hip. The incidence of hip dysplasia in the children of women surveyed was 45/1,000 which is 5 times the Norwegian incidence. CONCLUSION: Pelvic joint syndrome nearly always follows pelvic girdle relaxation of pregnancy and may have prolonged debilitating effects which do not respond long term to current therapies. The incidence of developmental dysplasia of the hip in the children of these women was high. A genetic susceptibility to joint dysfunction in both mother and fetus, possibly due to an aberration of relaxin physiology, is surmized. Identification of possible relaxin receptor changes in affected joints is a hypothesis worthy of testing with a view to the design of selective relaxin receptor modulators in pregnancy. PMID- 9348255 TI - Oxygen free radical activity in the second stage of labor. AB - OBJECTIVE: To assess fetal cellular injury arising from oxygen free radical activity in relation to the duration of the second stage of labor. PATIENTS AND METHODS: Cord arterial pH, malondialdehyde and organic hydroperoxides levels were determined following vaginal delivery of 326 term singleton pregnancies. Of these, 35 (11%) received epidural analgesia. The length of the second stage was recorded from the time of full dilatation to delivery. RESULTS: Arterial cord pH, malondialdehyde and organic hydroperoxides were significantly correlated with duration of second stage of labor (r=-0.4492; r=0.2542; r=0.2244; respectively, p<0.001). The association of lipid peroxidation products with second stage duration was independent of cord pH. This correlation was unchanged when cases of operative delivery were excluded. However, the relationship lost statistical significance amongst cases who received epidurals. CONCLUSIONS: The duration of the second stage is correlated with raised reactive oxygen species-derived lipid peroxidation products. Caution should be exercised in the management of prolonged second stage of labor. PMID- 9348256 TI - The effect of a normal vaginal delivery on anal function. AB - BACKGROUND: To determine the effect of an uncomplicated vaginal delivery on anal sphincter function in primiparous women. METHODS: In a prospective study, anal manometry was performed prenatally and at 4-6 weeks postnatally in 18 primiparous women (11 undergoing vaginal delivery; seven having Cesarean section). No patient had any evidence of anal sphincter damage after delivery as determined by anal ultrasound and pudendal nerve terminal motor latency. RESULTS: Vaginal delivery was associated with a significant reduction in the squeeze pressure (SP: prenatal 269 cm H2O vs postnatal 204 cm H2O; p=0.004) but not the resting pressure (RP: prenatal 96 cm H2O vs postnatal 86 cm H2O; p=0.075). Cesarean section was not associated with any significant change in anal pressures. CONCLUSION: A normal vaginal delivery with no evidence of sphincter injury was associated with a significant effect on anal function when measured 4-6 weeks postnatally. PMID- 9348257 TI - Diagnosis of post-partum ovarian vein thrombophlebitis by color Doppler ultrasonography: about 10 cases. AB - BACKGROUND: Post-partum ovarian vein thrombosis is often overlooked or mistaken for other complications such as endometritis. Color Doppler ultrasonography is a very good diagnostic method when properly indicated and correctly interpreted according to clinical data. METHODS: This study reports ten cases that were retrospectively studied, during which color Doppler ultrasonography was used. The clinical signs and the results are reviewed. RESULTS: The lesions were clearly visualized in eight of the ten cases; one of the two failures resulted from a methodological fault (uninterpretable result); the other one was due to the lack of experience of the operator and nonrecognition of the clinical signs. Thrombosis appears as a hypoechogenic and tubular image. This type of examination is particularly indicated in the presence of certain clinical signs that were observed in our cases: fever and iliac pain are the main precursor signs, often associated with abdominal meteorism and slow digestive transit; provoked cul-de sac pain during vaginal probing was the only constant sign, sometimes associated with painful swelling. PMID- 9348258 TI - Epstein-Barr virus DNA in the uterine cervix of teenage girls. AB - OBJECTIVES: To: (i) evaluate longitudinally the prevalence of Epstein-Barr virus (EBV) DNA in the cervix of healthy teenage girls, (ii) relate the presence of cervical EBV DNA to virginity or sexual experience, and (iii) relate the occurrence of cervical EBV DNA to the presence of specific IgG antibodies to EBV virus capsid antigen (EBV-VCA) in serum and to signs of genital infection. MATERIAL AND METHODS: Thirty-six teenage girls were followed for 2 years between the ages of 16 and 18 years. A sexual history was taken and a gynecological examination was performed on each occasion. The presence of EBV DNA in the cervix and of EBV VCA antibodies in serum was determined on each occasion. RESULTS: Coitus debut was reported by 23/36 girls (64%) and by 31/36 (86%) at 16- and 18 years of age, respectively. Two girls (only one with sexual debut) harbored EBV DNA in the cervix at 16 years of age. At the age of 18, no EBV DNA was found in these two girls, but another three girls carried EBV DNA in the cervix. All were sexually active and reported 1, 4 and 7 life-time sexual partners respectively. Serum EBV-VCA antibodies were found in 83% of the 16-year old girls and in 89% of the 18-year old girls (no significant difference between sexually experienced and virginal girls at either age). All the girls with cervical EBV DNA had antibodies against EBV-VCA. None of the girls with EBV DNA were found to carry HPV DNA or have a chlamydial infection in the cervix at any time during the study. There was no significant difference in the number of girls with a cervix secretion predominated by leucocytes between girls with positive and negative cervical EBV DNA samples. CONCLUSIONS: We conclude that among these healthy teenage girls the non-sexual route of transmission of EBV is more plausible than the sexual one. PMID- 9348260 TI - Transvaginal ultrasound, endometrial cytology sampled by Gynoscann and histology obtained by Uterine Explora Curette compared to the histology of the uterine specimen. A prospective study in pre- and postmenopausal women undergoing elective hysterectomy. AB - BACKGROUND: The purpose of this study was to evaluate the diagnostic value of transvaginal ultrasound measurement of endometrial thickness, cytology obtained by Gynoscann, and histology of the endometrium sampled by Uterine Explora Curette compared with histology of the uterine specimen as the gold standard. METHODS: Consecutive patients admitted for hysterectomy had transvaginal ultrasound, sampling by Gynoscann, and Uterine Explora Curette done just before surgery, after informed consent. RESULTS: A total of 181 women entered the study. Sixteen had endometrial cancer, seven had atypical hyperplasia and nine had complex hyperplasia. A total of 168 patients had a transvaginal ultrasound done. At a cutoff limit of 4mm (endometrial thickness of 4mm or less indicating normal endometrium), the sensitivity was 90.3%, the specificity 24.8%, the positive predictive value 21.4% and the negative predictive value 91.9%. One endometrial cancer, one atypical and one complex hyperplasia were missed. The Gynoscann method showed a sensitivity of 62.5%, a specificity of 94.0%, a positive predictive value of 69.0% and a negative predictive value of 92.1%. Two cancers, three atypical and six complex hyperplasia were missed. The Uterine Explora Curette showed a sensitivity of 90.6%, a specificity of 100.0%, a positive predictive value of 100.0% and a negative predictive value of 98.0%. One endometrial cancer and two complex hyperplasia were missed. CONCLUSION: Transvaginal ultrasound is a reliable method in excluding endometrial pathology. The Uterine Explora Curette was superior to Gynoscann in diagnosing neoplasia of the endometrium. It was found to have the same diagnostic accuracy as conventional dilatation and curettage. PMID- 9348259 TI - Ovarian volume in gynecologically healthy women using no contraception, or using IUD or oral contraception. AB - OBJECTIVE: The aim of this study was to determine the ovarian volume by transvaginal ultrasonography in a gynecologically healthy population of women using no contraception, using intrauterine contraceptive device, or using oral contraceptive. MATERIALS AND METHOD: The study had a cross-sectional design. The ovaries of 428 women aged 1445 who contacted the family planning clinic in the county of Funen were examined. Most of the statistical analyses were carried out using standard techniques. However polynominal regression analysis was used to model ovarian volumes as a function of the day of cycle. RESULTS: No differences between the volumes of the right and the left ovary were found in any of the groups. Significant differences were found between the ovarian volumes of the three groups. The ovarian volumes were found to be largest in women using intrauterine contraceptive device, lesser in women using no contraception and smallest in women using oral contraception. A significant difference was found of the ovarian volumes throughout the menstrual cycle in women who were not using oral contraception. The ovarian volumes did not change throughout the menstrual cycle in women using oral contraception. In women not using oral contraception the largest ovary increased in volume from the start of the cycle to day 19, thereafter the volume declined. No evidence of any change of volume over the menstrual cycle was found in the smallest ovary and, for women using oral contraception, both ovaries. There was no correlation between age, height, weight, parity, and ovarian volume in any of the groups. CONCLUSION: The ovarian volumes, in gynecologically healthy women using intrauterine contraceptive device, are larger than in women using no contraception. It appears that oral contraception reduces the volumes of both ovaries in all phases of the menstrual cycle to equal levels. PMID- 9348261 TI - Laparoscopic instillation of hyperosmolar glucose vs. expectant management of tubal pregnancies with serum hCG < or = 2500 mIU/mL. AB - OBJECTIVE: To compare expectant management with local instillation of 50% glucose solution for tubal pregnancies with a serum hCG level < or = 2500 mIU/mL. DESIGN: Prospective, non-randomized, comparative clinical study. SETTING: Two university departments. PATIENTS: One hundred and twenty-eight patients with laparoscopically-confirmed tubal pregnancy and serum hCG < or = 2500 mIU/mL. INTERVENTIONS: Eighty patients in Graz were treated with laparoscopic instillation of 50% glucose solution and 48 patients in Turku were followed expectantly. MAIN OUTCOME MEASURES: Resolution of hCG excretion, need for further interventions. RESULTS: Seventy-four of the 80 patients (92%) in the glucose group (32 of 33 with an initial hCG < or = 250 mIU/mL and 42 of 47 with hCG 251 2500 mIU/mL) and 36 of 48 (75%) patients in the expectant group (19 of 23 with an initial hCG < or = 250 mIU/mL and 17 of 25 with hCG 251-2500 mIU/mL) had resolution of the pregnancy with no further intervention (p=0.008, chi-square test, odds ratio 0.24). CONCLUSIONS: Glucose instillation is superior to expectant management for patients with early tubal pregnancy. PMID- 9348262 TI - Corpus cavernosum-like tumor of the vulva. PMID- 9348263 TI - Uterine metastasis from gastric cancer. PMID- 9348265 TI - Amniotic fluid embolism. Report of two cases with coagulation disorder. PMID- 9348266 TI - Decreased parathyroid hormone serum levels in pregnant women with first-trimester vaginal bleeding. PMID- 9348264 TI - Sonographic findings in a case report of benign leiomyomatosis peritonealis disseminata. PMID- 9348267 TI - Effects of hemodilution on splanchnic perfusion and hepatorenal function. I. Splanchnic perfusion. AB - Perfusion of intestinal organs increases in response to acute normovolemic hemodilution (ANH). However, detailed studies on distribution of regional splanchnic organ perfusion during ANH are lacking. We therefore carried out this study to test the hypothesis that ANH does not cause disturbance of physiologic patterns of regional splanchnic organ blood flow. After governmental permission, 22 anesthetized dogs were instrumented to allow invasive hemodynamic measurements and intracardial injection of radioactive microspheres (diameter 15 micro m) for determination of regional organ perfusion. Measurements were made at baseline (hematocrit 37 +/- 3%) and after ANH with 6% hydroxyethyl starch (mol. wt. 200000 / 0.5) to hct 20 +/- 1%. After completion of the protocol, splanchnic organs were removed and dissected into small samples according to anatomical and functional principles. Regional perfusion was determined based on the microsphere content of each sample. Hepatic, intestinal, and pancreatic blood flow increased with ANH. Hepatic arterial blood flow rose by 86%, whereas portal venous perfusion increased by 28%. Small intestine mucosal perfusion was augmented by 68% while the non-mucosal tissue compartment of the gut wall received 32% more blood flow after ANH which is in proportion to the increase in cardiac index after ANH. This redistribution of intestinal flow might be the basis for the preservation of tissue oxygenation during moderate isovolemic anemia. PMID- 9348268 TI - Effects of hemodilution on splanchnic perfusion and hepatorenal function. II. Renal perfusion and hepatorenal function. AB - Hepatorenal perfusion and function were assxssed in 22 dogs undergoing acute normovolemic hemodilution (ANH) to a hematocrit (Hct) of 20% using 6% hydroxyethyl starch (200.000/0.5) as the diluent. Organ perfusion was determined with the radioactive microspheres method. Renal function was assessed by urinary output, creatinine clearance and fractional sodium excretion. Blood volume as well as hepatic function were derived from indocyanine green (ICG) dilution kinetics. Hepatocellular integrity was determined by serum enzymatic activity of glutamate-oxalacetate-transaminase (GOT) and glutamate-pyruvate- transaminase (GPT). ANH to Hct 20% did not change blood volume and mean aortic pressure, while heart rate was slightly elevated (p<0.05) by 5 beats per minute and cardiac output increased by 29% (p<0.05). In contrast to the liver, where arterial and portal venous blood flow increased (86% and 28%, respectively; p<0.05), total renal blood flow as well as intraorgan distribution of renal blood flow remained unchanged post-ANH. While creatinine clearance remained unchanged following ANH, urinary output and fractional urinary excretion increased (p<0.05). In response to enhanced hepatic blood flow after ANH, intravascular half-life of ICG was reduced (p<0.05) and ICG clearance increased (p<0.05). Serum enzymatic activity of GPT decreased upon ANH (p<0.05), while GOT activity remained unchanged. ANH to a Hct 20% does not impair hepatorenal function. Increased urinary output points out the necessity for proper adjustment of crystalloid infusion to maintain normal intravascular volume and avoid hypovolemia and the associated risk of tissue hypoxia. PMID- 9348269 TI - Disappearing bone disease (Gorham-stout disease): report of a case with a follow up of 48 years. AB - A patient with osteolysis of the right hand after a metatarsal fracture at the age of 12 years is reported. The osteolysis progressed until the age of 21 years and was stable until the age of 59 years, when the patient died from a metastatic colon cancer. The article discusses the clinical, radiographic, and histologic features, and prognosis of idiopathic osteolysis (Gorham-Stout disease). Gorham Stout disease is characterized by a non-familial, histological benign vascular proliferation producing lysis of the bone. The therapeutic options of Gorham Stout disease is limited; radiotherapy has been used with success in single cases. Usually, the disease undergoes spontaneous arrest, as it occurred in our patient. PMID- 9348270 TI - Renal function and renotropic effects of secretin in cystic fibrosis. AB - In ten cystic fibrosis patients and nine age-matched controls, renal function was determined before and after infusion of secretin. Under baseline conditions creatinine excretion and clearance were significantly elevated, exclusively due to those patients who were homozygous for the DF508 mutation (153 vs 132 ml/min*1.73m2), whereas the glomerular filtration rate, measured by inulin clearance showed no difference. Renal plasma flow and the fractional reabsorption rates of electrolytes were similar in patients and controls. During secretin infusion renal plasma flow increased and the fractional reabsorption rates of electrolytes decreased in both groups. The patients had a increased metabolic clearance (2900 vs 1660 ml/min*m2) and endogenous production rate (9,9 vs 2,5 pmol/min*m2) of of secretin. In conclusion global renal function and electrolyte handling, in particular chloride permeability, are unchanged in cystic fibrosis. Individuals expressing the DF508 genotype showed a selective elevation of creatinine excretion and clearance. The secretion and metabolic clearance of secretin are increased in cystic fibrosis. PMID- 9348271 TI - Is the brain death related endocrine dysfunction an indication for hormonal substitution therapy in the early period ? AB - Experimental studies in animals have suggested that brain death (BD) -- related endocrinological dysregulations lead to a significant depression of cardiac pump and muscle function, however, the discussion about the relative extent of this influence remains controversal. The aim of the present study was to assess in an open chest animal model the short time course (5 hours) of hormonal (epinephrine, norepinephrine, T3, T4, ACTH, cortisol, insuline) and metabolic (glucose, lactate) changes in 10 brain dead dogs with special respect to the hemodynamic stability and myocardial pump function. After the onset of BD the concentrations of all hormonal parameters showed a significant decrease. Despite these changes, and in contrast to other studies, an adequate pump function (filling pressures, cardiac output) and muscle function (LVdp/dt) could be maintained by exclusive volume substitution without the use of hormonal or pressor agents. We conclude that in the present model a sufficient pump and muscle function can be maintained by adequate volume substitution, exclusively. The significant fall in adrenal and thyroid hormones had no direct effects on heart functional parameters in the first 5 hours after experimental BD induction. PMID- 9348272 TI - Diabetes mellitus as a clinical model of the process of aging. PMID- 9348273 TI - Treatment of macular degeneration, according to Bangerter. AB - Age-related macular degeneration (AMD) is a common cause of visual loss among elderly patients. Although some risk factors have been determined, the ultimate cause of the disease is not known. For a long time, therapeutic nihilism has been the rule among ophthalmologists confronted with such patients. Bangerter has not shared this attitude, especially since the time that he incidentally discovered, more than 40 years ago, the beneficial effects of radiotherapy, in discouraging the growth of new vessels at the posterior pole of the eye. A variety of approaches are combined and used by Bangerter in the treatment of the different types of AMD, including retrobulbar injections of either vasodilating medications (in the dry - or atrophic - type) or corticosteroids (in the wet - or exudative - type), general medical measures aimed at improving metabolic and vascular functions such as supplementation with trace elements, antioxidants, and vitamins; ozone therapy; advice to increase physical fitness, improve nutrition, and abstain from smoking; and protection from excessive light exposure. Being convinced of the usefulness of his type of combination treatment, he has always rejected undertaking controlled clinical trials, of only single aspects of the therapy, as unethical and invalid. For this reason, scientific journals have not proven cooperative in several attempts at publishing his results, as collected in retrospective surveys. Recently, however, some of the several approaches combined by Bangerter in treating AMD have been pronounced effective by other investigators. We present here an overview of his treatment approaches, as few people are aware of them, to clear up misconceptions and to set records straight. PMID- 9348274 TI - Intestinal adaptation and amino acid transport following massive enterectomy. AB - Morphological and physiological adaptation in residual small intestine occurs after massive enterectomy and is influenced significantly by different growth factors and hormones. The mechanism of adaptation occurs through hypertrophy and hyperplasia as well as nutrient transporter changes. These transporters are classified into different classes dependent on its biological properties. The adaptation process evolves over time and different nutrient absorption profiles occur at different postoperative stages. There is an initial decrease in amino acid transport after resection followed by a return to approximately normal levels. Glucose also follows a similar pattern of changes but returns to normal later than amino acids. The time course of these changes are different for different animals with rat adaptation being much faster than rabbit. Growth hormone (GH) induces increased amino acid transport during this adaptation period, however, appears not to affect small intestine hypertrophy or hyperplasia. The increase in transport occurs via an increase in transport numbers rather than affinity. Epidermal growth factor (EGF) also increases amino acid transport in postoperative animals. Its advantage is it is orally stable when given with a protease inhibitor. EGF also reverses the down-regulating effects of the somatostatin analogue Octreotide (SMS) post resection. EGF in combination with GH has additive effects. However, the effects of the growth factors are site specific. GH and EGF combination therapy significantly increased alanine and arginine transport in distal small bowel after 70 % enterectomy but not in the proximal small bowel. The same combination increases leucine and glutamine transport in the proximal small intestine only. Understanding the specific changes that occur with these therapies may improve quality of life for patients and also reduce that need for total parenteral nutrition. PMID- 9348275 TI - Integrins, oncogenes, and anchorage independence. PMID- 9348276 TI - Mapping and use of a sequence that targets DNA ligase I to sites of DNA replication in vivo. AB - The mammalian nucleus is highly organized, and nuclear processes such as DNA replication occur in discrete nuclear foci, a phenomenon often termed "functional organization" of the nucleus. We describe the identification and characterization of a bipartite targeting sequence (amino acids 1-28 and 111-179) that is necessary and sufficient to direct DNA ligase I to nuclear replication foci during S phase. This targeting sequence is located within the regulatory, NH2 terminal domain of the protein and is dispensable for enzyme activity in vitro but is required in vivo. The targeting domain functions position independently at either the NH2 or the COOH termini of heterologous proteins. We used the targeting sequence of DNA ligase I to visualize replication foci in vivo. Chimeric proteins with DNA ligase I and the green fluorescent protein localized at replication foci in living mammalian cells and thus show that these subnuclear functional domains, previously observed in fixed cells, exist in vivo. The characteristic redistribution of these chimeric proteins makes them unique markers for cell cycle studies to directly monitor entry into S phase in living cells. PMID- 9348278 TI - Ca2+ homeostasis in the endoplasmic reticulum: coexistence of high and low [Ca2+] subcompartments in intact HeLa cells. AB - Two recombinant aequorin isoforms with different Ca2+ affinities, specifically targeted to the endoplasmic reticulum (ER), were used in parallel to investigate free Ca2+ homeostasis in the lumen of this organelle. Here we show that, although identically and homogeneously distributed in the ER system, as revealed by both immunocytochemical and functional evidence, the two aequorins measured apparently very different concentrations of divalent cations ([Ca2+]er or [Sr2+]er). Our data demonstrate that this contradiction is due to the heterogeneity of the [Ca2+] of the aequorin-enclosing endomembrane system. Because of the characteristics of the calibration procedure used to convert aequorin luminescence into Ca2+ concentration, the [Ca2+]er values obtained at steady state tend, in fact, to reflect not the average ER values, but those of one or more subcompartments with lower [Ca2+]. These subcompartments are not generated artefactually during the experiments, as revealed by the dynamic analysis of the ER structure in living cells carried out by means of an ER-targeted green fluorescent protein. When the problem of ER heterogeneity was taken into account (and when Sr2+ was used as a Ca2+ surrogate), the bulk of the organelle was shown to accumulate free [cation2+]er up to a steady state in the millimolar range. A theoretical model, based on the existence of multiple ER subcompartments of high and low [Ca2+], that closely mimics the experimental data obtained in HeLa cells during accumulation of either Ca2+ or Sr2+, is presented. Moreover, a few other key problems concerning the ER Ca2+ homeostasis have been addressed with the following conclusions: (a) the changes induced in the ER subcompartments by receptor generation of InsP3 vary depending on their initial [Ca2+]. In the bulk of the system there is a rapid release whereas in the small subcompartments with low [Ca2+] the cation is simultaneously accumulated; (b) stimulation of Ca2+ release by receptor-generated InsP3 is inhibited when the lumenal level is below a threshold, suggesting a regulation by [cation2+]er of the InsP3 receptor activity (such a phenomenon had already been reported, however, but only in subcellular fractions analyzed in vitro); and (c) the maintenance of a relatively constant level of cytosolic [Ca2+], observed when the cells are incubated in Ca2+ free medium, depends on the continuous release of the cation from the ER, with ensuing activation in the plasma membrane of the channels thereby regulated (capacitative influx). PMID- 9348277 TI - Targeting of NH2-terminal-processed microsomal protein to mitochondria: a novel pathway for the biogenesis of hepatic mitochondrial P450MT2. AB - Cytochrome P4501A1 is a hepatic, microsomal membrane-bound enzyme that is highly induced by various xenobiotic agents. Two NH2-terminal truncated forms of this P450, termed P450MT2a and MT2b, are also found localized in mitochondria from beta-naphthoflavone-induced livers. In this paper, we demonstrate that P4501A1 has a chimeric NH2-terminal signal that facilitates the targeting of the protein to both the ER and mitochondria. The NH2-terminal 30-amino acid stretch of P4501A1 is thought to provide signals for ER membrane insertion and also stop transfer. The present study provides evidence that a sequence motif immediately COOH-terminal (residues 33-44) to the transmembrane domain functions as a mitochondrial targeting signal under both in vivo and in vitro conditions, and that the positively charged residues at positions 34 and 39 are critical for mitochondrial targeting. Results suggest that 25% of P4501A1 nascent chains, which escape ER membrane insertion, are processed by a liver cytosolic endoprotease. We postulate that the NH2-terminal proteolytic cleavage activates a cryptic mitochondrial targeting signal. Immunofluorescence microscopy showed that a portion of transiently expressed P4501A1 is colocalized with the mitochondrial specific marker protein cytochrome oxidase subunit I. The mitochondrial associated MT2a and MT2b are localized within the inner membrane compartment, as tested by resistance to limited proteolysis in both intact mitochondria and mitoplasts. Our results therefore describe a novel mechanism whereby proteins with chimeric signal sequence are targeted to the ER as well as to the mitochondria. PMID- 9348279 TI - The number and location of glycans on influenza hemagglutinin determine folding and association with calnexin and calreticulin. AB - Calnexin and calreticulin are homologous molecular chaperones that promote proper folding, oligomeric assembly, and quality control of newly synthesized glycoproteins in the endoplasmic reticulum (ER). Both are lectins that bind to substrate glycoproteins that have monoglucosylated N-linked oligosaccharides. Their binding to newly translated influenza virus hemagglutinin (HA), and various mutants thereof, was analyzed in microsomes after in vitro translation and expression in live CHO cells. A large fraction of the HA molecules was found to occur in ternary HA- calnexin-calreticulin complexes. In contrast to calnexin, calreticulin was found to bind primarily to early folding intermediates. Analysis of HA mutants with different numbers and locations of N-linked glycans showed that although the two chaperones share the same carbohydrate specificity, they display distinct binding properties; calreticulin binding depends on the oligosaccharides in the more rapidly folding top/hinge domain of HA whereas calnexin is less discriminating. Calnexin's binding was reduced if the HA was expressed as a soluble anchor-free protein rather than membrane bound. When the co- and posttranslational folding and trimerization of glycosylation mutants was analyzed, it was observed that removal of stem domain glycans caused accelerated folding whereas removal of the top domain glycans (especially the oligosaccharide attached to Asn81) inhibited folding. In summary, the data established that individual N-linked glycans in HA have distinct roles in calnexin/calreticulin binding and in co- and posttranslational folding. PMID- 9348280 TI - Mechanism of the facilitation of PC2 maturation by 7B2: involvement in ProPC2 transport and activation but not folding. AB - Among the members of the prohormone convertase (PC) family, PC2 has a unique maturation pattern: it is retained in the ER for a comparatively long time and its propeptide is cleaved in the TGN/ secretory granules rather than in the ER. It is also unique by its association with the neuroendocrine protein 7B2. This interaction results in the facilitation of proPC2 maturation and in the production of activatable proPC2 from CHO cells. In the present study, we have investigated the mechanism of this interaction. ProPC2 binds 7B2 in the ER, but exits this compartment much more slowly than 7B2. We found that proPC2 was also slow to acquire the capacity to bind 7B2, whereas 7B2 could bind proPC2 rapidly after synthesis. This indicated that proPC2 folding was the limiting step in the formation of the complex. Indeed, sensitivity of native proPC2 to N-glycanase F digestion and inhibition of proPC2 folding supported the notion that 7B2 is not involved in the early steps of proPC2 folding, and that proPC2 must fold before binding 7B2. Under experimental conditions that prevent propeptide cleavage, 7B2 expression increased proPC2 transport to the Golgi. This increase exhibited the same kinetics as the facilitation of the removal of the propeptide. Finally, proPC2 activation could be reconstituted in Golgi- enriched subcellular fractions. In vitro, 7B2 was required for proPC2 activation at an acidic pH. Taken together, our results demonstrate that rather than promoting proPC2 folding, 7B2 acts as a helper protein involved in proPC2 transport and is required in the proPC2 activation process. We propose, therefore, that 7B2 stabilizes proPC2 in a conformation already competent for these two events. PMID- 9348281 TI - Major histocompatibility complex class II compartments in human and mouse B lymphoblasts represent conventional endocytic compartments. AB - In most human and mouse antigen-presenting cells, the majority of intracellular major histocompatibility complex (MHC) class II molecules resides in late endocytic MHC class II compartments (MIICs), thought to function in antigen processing and peptide loading. However, in mouse A20 B cells, early endocytic class II-containing vesicles (CIIVs) have been reported to contain most of the intracellular MHC class II molecules and have also been implicated in formation of MHC class II-peptide complexes. To address this discrepancy, we have studied in great detail the endocytic pathways of both a human (6H5.DM) and a mouse (A20.Ab) B cell line. Using quantitative immunoelectron microscopy on cryosections of cells that had been pulse-chased with transferrin-HRP or BSA-gold as endocytic tracers, we have identified up to six endocytic subcompartments including an early MIIC type enriched in invariant chain, suggesting that it serves as an important entrance to the endocytic pathway for newly synthesized MHC class II/invariant chain complexes. In addition, early MIICs represented the earliest endocytic compartment containing MHC class II- peptide complexes, as shown by using an antibody against an abundant endogenous class II-peptide complex. The early MIIC exhibited several though not all of the characteristics reported for the CIIV and was situated just downstream of early endosomes. We have not encountered any special class II-containing endocytic structures besides those normally present in nonantigen-presenting cells. Our results therefore suggest that B cells use conventional endocytic compartments rather than having developed a unique compartment to accomplish MHC class II presentation. PMID- 9348282 TI - Phorbol esters and SDF-1 induce rapid endocytosis and down modulation of the chemokine receptor CXCR4. AB - The chemokine receptor CXCR4 is required, together with CD4, for entry by some isolates of HIV-1, particularly those that emerge late in infection. The use of CXCR4 by these viruses likely has profound effects on viral host range and correlates with the evolution of immunodeficiency. Stromal cell-derived factor-1 (SDF-1), the ligand for CXCR4, can inhibit infection by CXCR4-dependent viruses. To understand the mechanism of this inhibition, we used a monoclonal antibody that is specific for CXCR4 to analyze the effects of phorbol esters and SDF-1 on surface expression of CXCR4. On human T cell lines SupT1 and BC7, CXCR4 undergoes slow constitutive internalization (1.0% of the cell surface pool/min). Addition of phorbol esters increased this endocytosis rate >6-fold and reduced cell surface CXCR4 expression by 60 to 90% over 120 min. CXCR4 was internalized through coated pits and coated vesicles and subsequently localized in endosomal compartments from where it could recycle to the cell surface after removal of the phorbol ester. SDF-1 also induced the rapid down modulation (half time approximately 5 min) of CXCR4. Using mink lung epithelial cells expressing CXCR4 and a COOH-terminal deletion mutant of CXCR4, we found that an intact cytoplasmic COOH-terminal domain was required for both PMA and ligand-induced CXCR4 endocytosis. However, experiments using inhibitors of protein kinase C indicated that SDF-1 and phorbol esters trigger down modulation through different cellular mechanisms. SDF-1 inhibited HIV-1 infection of mink cells expressing CD4 and CXCR4. The inhibition of infection was less efficient for CXCR4 lacking the COOH terminal domain, suggesting at least in part that SDF-1 inhibition of virus infection was mediated through ligand-induced internalization of CXCR4. Significantly, ligand induced internalization of CXCR4 but not CD4, suggesting that CXCR4 and CD4 do not normally physically interact on the cell surface. Together these studies indicate that endocytosis can regulate the cell-surface expression of CXCR4 and that SDF-1-mediated down regulation of cell-surface coreceptor expression contributes to chemokine-mediated inhibition of HIV infection. PMID- 9348283 TI - Homotypic lysosome fusion in macrophages: analysis using an in vitro assay. AB - Lysosomes are dynamic structures capable of fusing with endosomes as well as other lysosomes. We examined the biochemical requirements for homotypic lysosome fusion in vitro using lysosomes obtained from rabbit alveolar macrophages or the cultured macrophage-like cell line, J774E. The in vitro assay measures the formation of a biotinylated HRP-avidin conjugate, in which biotinylated HRP and avidin were accumulated in lysosomes by receptor-mediated endocytosis. We determined that lysosome fusion in vitro was time- and temperature-dependent and required ATP and an N-ethylmaleimide (NEM)-sensitive factor from cytosol. The NEM sensitive factor was NSF as purified recombinant NSF could completely replace cytosol in the fusion assay whereas a dominant-negative mutant NSF inhibited fusion. Fusion in vitro was extensive; up to 30% of purified macrophage lysosomes were capable of self-fusion. Addition of GTPgammas to the in vitro assay inhibited fusion in a concentration-dependent manner. Purified GDP-dissociation inhibitor inhibited homotypic lysosome fusion suggesting the involvement of rabs. Fusion was also inhibited by the heterotrimeric G protein activator mastoparan, but not by its inactive analogue Mas-17. Pertussis toxin, a Galphai activator, inhibited in vitro lysosome fusion whereas cholera toxin, a Galphas activator did not inhibit the fusion reaction. Addition of agents that either promoted or disrupted microtubule function had little effect on either the extent or rate of lysosome fusion. The high value of homotypic fusion was supported by in vivo experiments examining lysosome fusion in heterokaryons formed between cells containing fluorescently labeled lysosomes. In both macrophages and J774E cells, almost complete mixing of the lysosome labels was observed within 1-3 h of UV sendai-mediated cell fusion. These studies provide a model system for identifying the components required for lysosome fusion. PMID- 9348284 TI - Flexibility within myosin heads revealed by negative stain and single-particle analysis. AB - Electron microscopy of negatively stained myosin has previously revealed three discrete regions within the heads of the molecule. However, despite a probable resolution of approximately 2 nm, it is difficult to discern directly consistent details within these regions. This is due to variability in both head conformation and in staining. In this study, we applied single-particle image processing and classified heads into homogeneous groups. The improved signal-to noise ratio after averaging these groups reveals substantially improved detail. The image averages were compared to a model simulating negative staining of the atomic structure of subfragment-1 (S1). This shows that the three head regions correspond to the motor domain and the essential and regulatory light chains. The image averages were very similar to particular views of the S1 model. They also revealed considerable flexibility between the motor and regulatory domains, despite the molecules having been prepared in the absence of nucleotide. This flexibility probably results from rotation of the regulatory domain about the motor domain, where the relative movement of the regulatory light chain is up to 12 nm, and is most clearly illustrated in animated sequences (available at http://www.leeds.ac.uk/chb/muscle/myosinhead.htm l). The sharply curved conformation of the atomic model of S1 is seen only rarely in our data, with straighter heads being more typical. PMID- 9348286 TI - Tomographic three-dimensional reconstruction of insect flight muscle partially relaxed by AMPPNP and ethylene glycol. AB - Rigor insect flight muscle (IFM) can be relaxed without ATP by increasing ethylene glycol concentration in the presence of adenosine 5'-[beta'gamma- imido]triphosphate (AMPPNP). Fibers poised at a critical glycol concentration retain rigor stiffness but support no sustained tension ("glycol-stiff state"). This suggests that many crossbridges are weakly attached to actin, possibly at the beginning of the power stroke. Unaveraged three-dimensional tomograms of "glycol-stiff" sarcomeres show crossbridges large enough to contain only a single myosin head, originating from dense collars every 14.5 nm. Crossbridges with an average 90 degrees axial angle contact actin midway between troponin subunits, which identifies the actin azimuth in each 38.7-nm period, in the same region as the actin target zone of the 45 degrees angled rigor lead bridges. These 90 degrees "target zone" bridges originate from the thick filament and approach actin at azimuthal angles similar to rigor lead bridges. Another class of glycol PNP crossbridge binds outside the rigor actin target zone. These "nontarget zone" bridges display irregular forms and vary widely in axial and azimuthal attachment angles. Fitting the acto-myosin subfragment 1 atomic structure into the tomogram reveals that 90 degrees target zone bridges share with rigor a similar contact interface with actin, while nontarget crossbridges have variable contact interfaces. This suggests that target zone bridges interact specifically with actin, while nontarget zone bridges may not. Target zone bridges constitute only approximately 25% of the myosin heads, implying that both specific and nonspecific attachments contribute to the high stiffness. The 90 degrees target zone bridges may represent a preforce attachment that produces force by rotation of the motor domain over actin, possibly independent of the regulatory domain movements. PMID- 9348285 TI - Brush border myosin-I structure and ADP-dependent conformational changes revealed by cryoelectron microscopy and image analysis. AB - Brush border myosin-I (BBM-I) is a single-headed myosin found in the microvilli of intestinal epithelial cells, where it forms lateral bridges connecting the core bundle of actin filaments to the plasma membrane. Extending previous observations (Jontes, J.D., E.M. Wilson-Kubalek, and R.A. Milligan. 1995. Nature [Lond.]. 378:751-753), we have used cryoelectron microscopy and helical image analysis to generate three-dimensional (3D) maps of actin filaments decorated with BBM-I in both the presence and absence of 1 mM MgADP. In the improved 3D maps, we are able to see the entire light chain-binding domain, containing density for all three calmodulin light chains. This has enabled us to model a high resolution structure of BBM-I using the crystal structures of the chicken skeletal muscle myosin catalytic domain and essential light chain. Thus, we are able to directly measure the full magnitude of the ADP-dependent tail swing. The approximately 31 degrees swing corresponds to approximately 63 A at the end of the rigid light chain-binding domain. Comparison of the behavior of BBM-I with skeletal and smooth muscle subfragments-1 suggests that there are substantial differences in the structure and energetics of the biochemical transitions in the actomyosin ATPase cycle. PMID- 9348287 TI - Neurotrophin-3-enhanced nerve regeneration selectively improves recovery of muscle fibers expressing myosin heavy chains 2b. AB - The purpose of this study was to evaluate the effect of neurotrophin 3 (NT-3) enhanced nerve regeneration on the reinnervation of a target muscle. Muscle fibers can be classified according to their mechanical properties and myosin heavy chain (MHC) isoform composition. MHC1 containing slow-type and MHC2a or 2b fast-type fibers are normally distributed in a mosaic pattern, their phenotype dictated by motor innervation. After denervation, all fibers switch to fast-type MHC2b expression and also undergo atrophy resulting in loss of muscle mass. After regeneration, discrimination between fast and slow fibers returns, but the distribution and fiber size change according to the level of reinnervation. In this study, rat gastrocnemius muscles (ipsilateral and contralateral to the side of nerve injury) were collected up to 8 mo after nerve repair, with or without local delivery of NT-3. The phenotype changes of MHC1, 2a, and 2b were analyzed by immunohistochemistry, and fiber type proportion, diameter, and grouping were assessed by computerized image analysis. At 8 mo, the local delivery of NT-3 resulted in significant improvement in gastrocnemius muscle weight compared with controls (NT-3 group 47%, controls 39% weight of contralateral normal muscle; P < 0.05). NT-3 delivery resulted in a significant increase in the proportion (NT-3 43.3%, controls 35.7%; P < 0.05) and diameter (NT-3 87.8 micron, controls 70.8 micron; P < 0.05) of fast type 2b fibers after reinnervation. This effect was specific to type 2b fibers; no normalization was seen in other fiber types. This study indicates that NT-3-enhanced axonal regeneration has a beneficial effect on the motor target organ. Also, NT-3 may be specifically affecting a subset of motoneurons that determine type 2b muscle fiber phenotype. As NT-3 was topically applied to cut nerves, our data suggest a discriminating effect of the neurotrophin on neuro-muscular interaction. These results would imply that muscle fibers may be differentially responsive to other neurotrophic factors and indicate the potential clinical role of NT-3 in the prevention of muscle atrophy after nerve injury. PMID- 9348289 TI - A yeast mutant showing diagnostic markers of early and late apoptosis. AB - A Saccharomyces cerevisiae mutant in cell division cycle gene CDC48 shows typical markers of apoptosis: membrane staining with annexin V, indicating an exposure of phosphatidylserine at the outer layer of the cytoplasmic membrane; intense staining, using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method, indicating DNA fragmentation; and chromatin condensation and fragmentation. The coordinate occurrence of these events at different locations in the cell, which have no obvious connection except their relation to apoptosis, implies the presence of the molecular machinery performing the basic steps of apoptosis already in yeast. Saccharomyces cerevisiae may prove a suitable model to trace the roots of apoptosis. PMID- 9348288 TI - Mal3, the fission yeast homologue of the human APC-interacting protein EB-1 is required for microtubule integrity and the maintenance of cell form. AB - Through a screen designed to isolate novel fission yeast genes required for chromosome segregation, we have identified mal3+. The mal3-1 mutation decreased the transmission fidelity of a nonessential minichromosome and altered sensitivity to microtubule-destabilizing drugs. Sequence analysis revealed that the 35-kD Mal3 is a member of an evolutionary conserved protein family. Its human counterpart EB-1 was identified in an interaction screen with the tumour suppressor protein APC. EB-1 was able to substitute for the complete loss of the mal3+ gene product suggesting that the two proteins might have similar functions. Cells containing a mal3 null allele were viable but showed a variety of phenotypes, including impaired control of cell shape. A fusion protein of Mal3 with the Aequorea victoria green fluorescent protein led to in vivo visualization of both cytoplasmic and mitotic microtubule structures indicating association of Mal3 with microtubules. The absence of Mal3 protein led to abnormally short, often faint cytoplasmic microtubules as seen by indirect antitubulin immunofluorescence. While loss of the mal3+ gene product had no gross effect on mitotic spindle morphology, overexpression of mal3+ compromised spindle formation and function and led to severe growth inhibition and abnormal cell morphology. We propose that Mal3 plays a role in regulating the integrity of microtubules possibly by influencing their stability. PMID- 9348290 TI - Specific and redundant functions of retinoid X Receptor/Retinoic acid receptor heterodimers in differentiation, proliferation, and apoptosis of F9 embryonal carcinoma cells. AB - We have generated F9 murine embryonal carcinoma cells in which either the retinoid X receptor (RXR)alpha and retinoic acid receptor (RAR)alpha genes or the RXRalpha and RARgamma genes are knocked out, and compared their phenotypes with those of wild-type (WT), RXRalpha-/-, RARalpha-/-, and RARgamma-/- cells. RXRalpha-/-/ RARalpha-/- cells were resistant to retinoic acid treatment for the induction of primitive and parietal endodermal differentiation, as well as for antiproliferative and apoptotic responses, whereas they could differentiate into visceral endodermlike cells, as previously observed for RXRalpha-/- cells. In contrast, RXRalpha-/-/RARgamma-/- cells were defective for all three types of differentiation, as well as antiproliferative and apoptotic responses, indicating that RXRalpha and RARgamma represent an essential receptor pair for these responses. Taken together with results obtained by treatment of WT and mutant F9 cells with RAR isotype- and panRXR-selective retinoids, our observations support the conclusion that RXR/ RAR heterodimers are the functional units mediating the retinoid signal in vivo. Our results also indicate that the various heterodimers can exert both specific and redundant functions in differentiation, proliferation, and apoptosis. We also show that the functional redundancy exhibited between RXR isotypes and between RAR isotypes in cellular processes can be artifactually generated by gene knockouts. The present approach for multiple gene targeting should allow inactivation of any set of genes in a given cell. PMID- 9348291 TI - ERM (ezrin/radixin/moesin)-based molecular mechanism of microvillar breakdown at an early stage of apoptosis. AB - Breakdown of microvilli is a common early event in various types of apoptosis, but its molecular mechanism and implications remain unclear. ERM (ezrin/radixin/moesin) proteins are ubiquitously expressed microvillar proteins that are activated in the cytoplasm, translocate to the plasma membrane, and function as general actin filament/plasma membrane cross-linkers to form microvilli. Immunofluorescence microscopic and biochemical analyses revealed that, at the early phase of Fas ligand (FasL)-induced apoptosis in L cells expressing Fas (LHF), ERM proteins translocate from the plasma membranes of microvilli to the cytoplasm concomitant with dephosphorylation. When the FasL induced dephosphorylation of ERM proteins was suppressed by calyculin A, a serine/threonine protein phosphatase inhibitor, the cytoplasmic translocation of ERM proteins was blocked. The interleukin-1beta-converting enzyme (ICE) protease inhibitors suppressed the dephosphorylation as well as the cytoplasmic translocation of ERM proteins. These findings indicate that during FasL-induced apoptosis, the ICE protease cascade was first activated, and then ERM proteins were dephosphorylated followed by their cytoplasmic translocation, i.e., microvillar breakdown. Next, to examine the subsequent events in microvillar breakdown, we prepared DiO-labeled single-layered plasma membranes with the cytoplasmic surface freely exposed from FasL-treated or nontreated LHF cells. On single-layered plasma membranes from nontreated cells, ERM proteins and actin filaments were densely detected, whereas those from FasL-treated cells were free from ERM proteins or actin filaments. We thus concluded that the cytoplasmic translocation of ERM proteins is responsible for the microvillar breakdown at an early phase of apoptosis and that the depletion of ERM proteins from plasma membranes results in the gross dissociation of actin-based cytoskeleton from plasma membranes. The physiological relevance of this ERM protein-based microvillar breakdown in apoptosis will be discussed. PMID- 9348292 TI - Dismantling cell-cell contacts during apoptosis is coupled to a caspase-dependent proteolytic cleavage of beta-catenin. AB - Cell death by apoptosis is a tightly regulated process that requires coordinated modification in cellular architecture. The caspase protease family has been shown to play a key role in apoptosis. Here we report that specific and ordered changes in the actin cytoskeleton take place during apoptosis. In this context, we have dissected one of the first hallmarks in cell death, represented by the severing of contacts among neighboring cells. More specifically, we provide demonstration for the mechanism that could contribute to the disassembly of cytoskeletal organization at cell-cell adhesion. In fact, beta-catenin, a known regulator of cell-cell adhesion, is proteolytically processed in different cell types after induction of apoptosis. Caspase-3 (cpp32/apopain/yama) cleaves in vitro translated beta-catenin into a form which is similar in size to that observed in cells undergoing apoptosis. beta-Catenin cleavage, during apoptosis in vivo and after caspase-3 treatment in vitro, removes the amino- and carboxy-terminal regions of the protein. The resulting beta-catenin product is unable to bind alpha-catenin that is responsible for actin filament binding and organization. This evidence indicates that connection with actin filaments organized at cell cell contacts could be dismantled during apoptosis. Our observations suggest that caspases orchestrate the specific and sequential changes in the actin cytoskeleton occurring during cell death via cleavage of different regulators of the microfilament system. PMID- 9348293 TI - The amino-terminal domain of desmoplakin binds to plakoglobin and clusters desmosomal cadherin-plakoglobin complexes. AB - The desmosome is a highly organized plasma membrane domain that couples intermediate filaments to the plasma membrane at regions of cell-cell adhesion. Desmosomes contain two classes of cadherins, desmogleins, and desmocollins, that bind to the cytoplasmic protein plakoglobin. Desmoplakin is a desmosomal component that plays a critical role in linking intermediate filament networks to the desmosomal plaque, and the amino-terminal domain of desmoplakin targets desmoplakin to the desmosome. However, the desmosomal protein(s) that bind the amino-terminal domain of desmoplakin have not been identified. To determine if the desmosomal cadherins and plakoglobin interact with the amino-terminal domain of desmoplakin, these proteins were co-expressed in L-cell fibroblasts, cells that do not normally express desmosomal components. When expressed in L-cells, the desmosomal cadherins and plakoglobin exhibited a diffuse distribution. However, in the presence of an amino-terminal desmoplakin polypeptide (DP-NTP), the desmosomal cadherins and plakoglobin were observed in punctate clusters that also contained DP-NTP. In addition, plakoglobin and DP-NTP were recruited to cell cell interfaces in L-cells co-expressing a chimeric cadherin with the E-cadherin extracellular domain and the desmoglein-1 cytoplasmic domain, and these cells formed structures that were ultrastructurally similar to the outer plaque of the desmosome. In transient expression experiments in COS cells, the recruitment of DP-NTP to cell borders by the chimera required co-expression of plakoglobin. Plakoglobin and DP-NTP co-immunoprecipitated when extracted from L-cells, and yeast two hybrid analysis indicated that DP-NTP binds directly to plakoglobin but not Dsg1. These results identify a role for desmoplakin in organizing the desmosomal cadherin-plakoglobin complex and provide new insights into the hierarchy of protein interactions that occur in the desmosomal plaque. PMID- 9348295 TI - The guanine nucleotide exchange factor Tiam1 affects neuronal morphology; opposing roles for the small GTPases Rac and Rho. AB - The invasion-inducing T-lymphoma invasion and metastasis 1 (Tiam1) protein functions as a guanine nucleotide exchange factor (GEF) for the small GTPase Rac1. Differentiation-dependent expression of Tiam1 in the developing brain suggests a role for this GEF and its effector Rac1 in the control of neuronal morphology. Here we show that overexpression of Tiam1 induces cell spreading and affects neurite outgrowth in N1E-115 neuroblastoma cells. These effects are Rac dependent and strongly promoted by laminin. Overexpression of Tiam1 recruits the alpha 6 beta 1 integrin, a laminin receptor, to specific adhesive contacts at the cell periphery, which are different from focal contacts. Cells overexpressing Tiam1 no longer respond to lysophosphatidic acid- induced neurite retraction and cell rounding, processes mediated by Rho, suggesting that Tiam1-induced activation of Rac antagonizes Rho signaling. This inhibition can be overcome by coexpression of constitutively active RhoA, which may indicate that regulation occurs at the level of Rho or upstream. Conversely, neurite formation induced by Tiam1 or Rac1 is further promoted by inactivating Rho. These results demonstrate that Rac- and Rho-mediated pathways oppose each other during neurite formation and that a balance between these pathways determines neuronal morphology. Furthermore, our data underscore the potential role of Tiam1 as a specific regulator of Rac during neurite formation and illustrate the importance of reciprocal interactions between the cytoskeleton and the extracellular matrix during this process. PMID- 9348294 TI - The Ras target AF-6 interacts with ZO-1 and serves as a peripheral component of tight junctions in epithelial cells. AB - The dynamic rearrangement of cell-cell junctions such as tight junctions and adherens junctions is a critical step in various cellular processes, including establishment of epithelial cell polarity and developmental patterning. Tight junctions are mediated by molecules such as occludin and its associated ZO-1 and ZO-2, and adherens junctions are mediated by adhesion molecules such as cadherin and its associated catenins. The transformation of epithelial cells by activated Ras results in the perturbation of cell-cell contacts. We previously identified the ALL-1 fusion partner from chromosome 6 (AF-6) as a Ras target. AF-6 has the PDZ domain, which is thought to localize AF-6 at the specialized sites of plasma membranes such as cell-cell contact sites. We investigated roles of Ras and AF-6 in the regulation of cell-cell contacts and found that AF-6 accumulated at the cell-cell contact sites of polarized MDCKII epithelial cells and had a distribution similar to that of ZO-1 but somewhat different from those of catenins. Immunoelectron microscopy revealed a close association between AF-6 and ZO-1 at the tight junctions of MDCKII cells. Native and recombinant AF-6 interacted with ZO-1 in vitro. ZO-1 interacted with the Ras-binding domain of AF 6, and this interaction was inhibited by activated Ras. AF-6 accumulated with ZO 1 at the cell-cell contact sites in cells lacking tight junctions such as Rat1 fibroblasts and PC12 rat pheochromocytoma cells. The overexpression of activated Ras in Rat1 cells resulted in the perturbation of cell-cell contacts, followed by a decrease of the accumulation of AF-6 and ZO-1 at the cell surface. These results indicate that AF-6 serves as one of the peripheral components of tight junctions in epithelial cells and cell-cell adhesions in nonepithelial cells, and that AF-6 may participate in the regulation of cell-cell contacts, including tight junctions, via direct interaction with ZO-1 downstream of Ras. PMID- 9348296 TI - Activated phosphatidylinositol 3-kinase and Akt kinase promote survival of superior cervical neurons. AB - The signaling pathways that mediate the ability of NGF to support survival of dependent neurons are not yet completely clear. However previous work has shown that the c-Jun pathway is activated after NGF withdrawal, and blocking this pathway blocks neuronal cell death. In this paper we show that over-expression in sympathetic neurons of phosphatidylinositol (PI) 3-kinase or its downstream effector Akt kinase blocks cell death after NGF withdrawal, in spite of the fact that the c-Jun pathway is activated. Yet, neither the PI 3-kinase inhibitor LY294002 nor a dominant negative PI 3-kinase cause sympathetic neurons to die if they are maintained in NGF. Thus, although NGF may regulate multiple pathways involved in neuronal survival, stimulation of the PI 3-kinase pathway is sufficient to allow cells to survive in the absence of this factor. PMID- 9348297 TI - A sponge-like structure involved in the association and transport of maternal products during Drosophila oogenesis. AB - Localization of maternally provided RNAs during oogenesis is required for formation of the antero-posterior axis of the Drosophila embryo. Here we describe a subcellular structure in nurse cells and oocytes which may function as an intracellular compartment for assembly and transport of maternal products involved in RNA localization. This structure, which we have termed "sponge body," consists of ER-like cisternae, embedded in an amorphous electron-dense mass. It lacks a surrounding membrane and is frequently associated with mitochondria. The sponge bodies are not identical to the Golgi complexes. We suggest that the sponge bodies are homologous to the mitochondrial cloud in Xenopus oocytes, a granulo-fibrillar structure that contains RNAs involved in patterning of the embryo. Exuperantia protein, the earliest factor known to be required for the localization of bicoid mRNA to the anterior pole of the Drosophila oocyte, is highly enriched in the sponge bodies but not an essential structural component of these. RNA staining indicates that sponge bodies contain RNA. However, neither the intensity of this staining nor the accumulation of Exuperantia in the sponge bodies is dependent on the amount of bicoid mRNA present in the ovaries. Sponge bodies surround nuage, a possible polar granule precursor. Microtubules and microfilaments are not present in sponge bodies, although transport of the sponge bodies through the cells is implied by their presence in cytoplasmic bridges. We propose that the sponge bodies are structures that, by assembly and transport of included molecules or associated structures, are involved in localization of mRNAs in Drosophila oocytes. PMID- 9348298 TI - Phenotypic and functional separation of memory and effector human CD8+ T cells. AB - Human CD8+ memory- and effector-type T cells are poorly defined. We show here that, next to a naive compartment, two discrete primed subpopulations can be found within the circulating human CD8+ T cell subset. First, CD45RA-CD45R0(+) cells are reminiscent of memory-type T cells in that they express elevated levels of CD95 (Fas) and the integrin family members CD11a, CD18, CD29, CD49d, and CD49e, compared to naive CD8+ T cells, and are able to secrete not only interleukin (IL) 2 but also interferon gamma, tumor necrosis factor alpha, and IL 4. This subset does not exert cytolytic activity without prior in vitro stimulation but does contain virus-specific cytotoxic T lymphocyte (CTL) precursors. A second primed population is characterized by CD45RA expression with concomitant absence of expression of the costimulatory molecules CD27 and CD28. The CD8+CD45RA+CD27- population contains T cells expressing high levels of CD11a, CD11b, CD18, and CD49d, whereas CD62L (L-selectin) is not expressed. These T cells do not secrete IL-2 or -4 but can produce IFN-gamma and TNF-alpha. In accordance with this finding, cells contained within this subpopulation depend for proliferation on exogenous growth factors such as IL-2 and -15. Interestingly, CD8+CD45RA+CD27- cells parallel effector CTLs, as they abundantly express Fas-ligand mRNA, contain perforin and granzyme B, and have high cytolytic activity without in vitro prestimulation. Based on both phenotypic and functional properties, we conclude that memory- and effector-type T cells can be separated as distinct entities within the human CD8+ T cell subset. PMID- 9348299 TI - Molecular characterization and functional analysis of murine interleukin 4 receptor allotypes. AB - The murine interleukin 4 receptor (IL-4R) exists as a transmembrane protein transducing pleiotropic IL-4 functions, or as soluble (s)IL-4-binding molecule with potent immunoregulatory effects. In this study we identified and characterized a murine IL-4R allotype. Sequence analysis of the IL-4R cDNA of BALB/c mice revealed 18 base substitutions leading to three extracellular and five cytoplasmic amino acid changes when compared with the published IL-4R sequence of C57BL/6 mice. Analyses with allotype-specific mAbs revealed that AKR/J and SJL/J mice possess the newly identified BALB/c IL-4R allotype whereas the IL-4Rs of C3H, CBA, DBA-2, and FVB/N mice are identical to that of the C57BL/6 mouse. The extracellular Thr49 to Ile substitution abrogates one N glycosylation site in the naturally occurring BALB/c IL-4R as well as in the experimentally point mutated C57BL/6-T49I sIL-4R, and both molecules display a nearly threefold reduction in IL-4-neutralizing activity compared to the C57BL/6 sIL-4R. In line with this, a significantly enhanced dissociation rate of IL-4 was detected for the BALB/c IL-4R allotype by surface plasmon resonance and in radioligand binding studies with IL-4R-transfected cell lines. These findings suggest that the altered ligand binding behavior of the newly described IL-4R allotype may influence the IL-4 responsiveness, thus contributing to the diverse phenotypes of inbred mouse strains in IL-4-dependent diseases. PMID- 9348300 TI - The macrophage scavenger receptor type A is expressed by activated macrophages and protects the host against lethal endotoxic shock. AB - During gram-negative bacterial infections, lipopolysaccharide (LPS) stimulates primed macrophages (Mphi) to release inflammatory mediators such as tumor necrosis factor (TNF)-alpha, which can cause hypotension, organ failure, and often death. Several different receptors on Mphi have been shown to bind LPS, including the type A scavenger receptor (SR-A). This receptor is able to bind a broad range of polyanionic ligands such as modified lipoproteins and lipoteichoic acid of gram-positive bacteria, which suggests that SR-A plays a role in host defense. In this study, we used mice lacking the SR-A (SRKO) to investigate the role of SR-A in acquired immunity using a viable bacillus Calmette Guerin (BCG) infection model. We show that activated Mphi express SR-A and that this molecule is functional in assays of adhesion and endocytic uptake. After BCG infection, SRKO mice are able to recruit Mphi to sites of granuloma formation where they become activated and restrict BCG replication. However, infected mice lacking the SR-A are more susceptible to endotoxic shock and produce more TNF-alpha and interleukin-6 in response to LPS. In addition, we show that an antibody which blocks TNF-alpha activity reduces LPS-induced mortality in these mice. Thus SR-A, expressed by activated Mphi, plays a protective role in host defense by scavenging LPS as well as by reducing the release by activated Mphi of proinflammatory cytokines. Modulation of SR-A may provide a novel therapeutic approach to control endotoxic shock. PMID- 9348301 TI - Selection of antigen-specific T cells by a single IEk peptide combination. AB - In normal mice, major histocompatibility complex (MHC) proteins are bound to many different peptides, derived from the proteins of their host. In the thymus, the diversity of this collection of MHC + peptide ligands allows thymocytes bearing many different T cell receptors (TCRs) to mature by low avidity reactions between the MHC + peptide ligands and the thymocyte TCRs. To investigate this problem, the selection of T cells specific for a well-studied combination of MHC + peptide, IEk + moth cytochrome c 88-103 (MCC), was investigated. Mice were created that expressed IEk bound to a single peptide, either a variant of MCC in which a critical TCR contact residue, 99K, was changed to A, or a variant of a mouse hemoglobin 64-76 (Hb) peptide, 72A. IEk bound to the MCC variant caused the clonal deletion of some T cells specific for the IEk + MCC ligand; nevertheless, it also positively selected many T cells that could react with this ligand. Some of the TCRs on the selected T cells were related to those on cells from normal mice and some were not. IEk bound to the Hb variant, on the other hand, did not select any T cells which could react with IEk + MCC. These results demonstrate that although positive selection is a partially degenerate event, the sequence of the peptide involved in positive selection controls the selected repertoire. PMID- 9348302 TI - Antinuclear autoantibodies and lupus nephritis in transgenic mice expressing interferon gamma in the epidermis. AB - Systemic lupus erythematosus (SLE) is a potentially fatal non-organ-specific autoimmune disease that predominantly affects women. Features of the disease include inflammatory skin lesions and widespread organ damage caused by deposition of anti-dsDNA autoantibodies. The mechanism and site of production of these autoantibodies is unknown, but there is evidence that interferon (IFN) gamma plays a key role. We have used the involucrin promoter to overexpress IFN gamma in the suprabasal layers of transgenic mouse epidermis. There was no evidence of organ-specific autoimmunity, but transgenic animals produced autoantibodies against dsDNA and histones. Autoantibody levels in female mice were significantly higher than in male transgenic mice. Furthermore, there was IgG deposition in the glomeruli of all female mice and histological evidence of severe proliferative glomerulonephritis in a proportion of these animals. Our findings are consistent with a central role for the skin immune system, acting under the influence of IFN-gamma, in the pathogenesis of SLE. PMID- 9348303 TI - Subunit composition of pre-T cell receptor complexes expressed by primary thymocytes: CD3 delta is physically associated but not functionally required. AB - Maturation of immature CD4-CD8- (DN) thymocytes to the CD4+CD8+ (DP) stage of development is driven by signals transduced through a pre-T cell receptor (TCR) complex, whose hallmark is a novel subunit termed pre-T alpha (pT alpha). However, the precise role of pre-TCRs in mediating the DN to DP transition remains unclear. Moreover, progress in understanding pre-TCR function has been hampered thus far because previous attempts to demonstrate expression of pT alpha containing pre-TCRs on the surface of normal thymocytes have been unsuccessful. In this report, we demonstrate for the first time that pT alpha-containing pre TCR complexes are expressed at low levels on the surface of primary thymocytes and that these pre-TCR complexes comprise a disulfide-linked pT alpha-TCR-beta heterodimer associated not only with CD3-gamma and -epsilon, as previously reported, but also with zeta and delta. Interestingly, while CD3-delta is associated with the pre-TCR complex, it is not required for pre-TCR function, as evidenced by the generation of normal numbers of DP thymocytes in CD3-delta deficient mice. The fact that any of the signaling components of the pre-TCR are dispensable for pre-TCR function is indeed surprising, given that few pre-TCR complexes are actually expressed on the surface of primary thymocytes in vivo. Thus, pre-TCRs do not require the full array of TCR-associated signaling subunits (gamma, delta, epsilon, and zeta), possibly because pT alpha itself possesses signaling capabilities. PMID- 9348304 TI - Novel mutants define genes required for the expression of human histocompatibility leukocyte antigen DM: evidence for loci on human chromosome 6p. AB - We and others have shown that the products of the HLA-DM locus are required for the intracellular assembly of major histocompatibility complex class II molecules with cognate peptides for antigen presentation. HLA-DM heterodimers mediate the dissociation of invariant chain (Ii)-derived class II-associated Ii peptides (CLIP) from class II molecules and facilitate the loading of class II molecules with antigenic peptides. Here we describe novel APC mutants with defects in the formation of class II-peptide complexes. These mutants express class II molecules which are conformationally altered, and an aberrantly high percentage of these class II molecules are associated with Ii-derived CLIP. This phenotype resembles that of DM null mutants. However, we show that the defects in two of these new mutants do not map to the DM locus. Nevertheless, our evidence suggests that the antigen processing defective phenotype in these mutants results from deficient DM expression. These mutants thus appear to define genes in which mutations have differential effects on the expression of conventional class II molecules and DM molecules. Our data are most consistent with these factors mapping to human chromosome 6p. Previous data have suggested that the expression of DM and class II genes are coordinately regulated. The results reported here suggest that DM and class II can also be differentially regulated, and that this differential regulation has significant effects on class II-restricted antigen processing. PMID- 9348305 TI - Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility complex class II-restricted viral epitope. AB - Intramuscular and intracutaneous immunization with naked DNA can vaccinate animals to the encoded proteins, but the underlying mechanisms of antigen presentation are unclear. We used DNA that encodes an A/PR/8/34 influenza peptide for CD4 T cells and that elicits protective antiviral immunity. DNA-transfected, cultured muscle cells released the influenza polypeptide, which then could be presented on the major histocompatibility complex class II molecules of dendritic cells. When DNA was injected into muscles or skin, and antigen-presenting cells were isolated from either the draining lymph nodes or the skin, dendritic, but not B, cells presented antigen to T cells and carried plasmid DNA. We suggest that the uptake of DNA and/or the protein expressed by dendritic cells triggers immune responses to DNA vaccines. PMID- 9348306 TI - Requirements for both Rac1 and Cdc42 in membrane ruffling and phagocytosis in leukocytes. AB - Specific pathways linking heterotrimeric G proteins and Fcgamma receptors to the actin-based cytoskeleton are poorly understood. To test a requirement for Rho family members in cytoskeletal events mediated by structurally diverse receptors in leukocytes, we transfected the full-length human chemotactic peptide receptor in RAW 264.7 cells and examined cytoskeletal alterations in response to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP), colony stimulating factor-1 (CSF-1), IgG-coated particles, and phorbol 12-myristate 13 acetate (PMA). Expression of Rac1 N17, Cdc42 N17, or the GAP domain of n chimaerin inhibited cytoskeletal responses to FMLP and CSF-1, and blocked phagocytosis. Accumulation of F-actin- rich "phagocytic cups" was partially inhibited by expression of Rac1 N17 or Cdc42 N17. In contrast, PMA-induced ruffling was not inhibited by expression of Rac1 N17, but was blocked by expression of Cdc42 N17, indicating that cytoskeletal inhibition by these constructs was nonoverlapping. These results demonstrate differential requirements for Rho family GTPases in leukocyte motility, and indicate that both Rac1 and Cdc42 are required for Fcgamma receptor- mediated phagocytosis and for membrane ruffling mediated by structurally distinct receptors in macrophages. PMID- 9348307 TI - Interferon (IFN)-alpha activation of human blood mononuclear cells in vitro and in vivo for nitric oxide synthase (NOS) type 2 mRNA and protein expression: possible relationship of induced NOS2 to the anti-hepatitis C effects of IFN alpha in vivo. AB - Although researchers have noted high level activation of rodent mononuclear phagocytes for nitric oxide (NO) synthase type 2 (S2) expression and NO production with a variety of agents such as interferon (IFN) gamma and endotoxin, it has been difficult to demonstrate activation of human mononuclear phagocytes. The purpose of this study was to determine if IFN-alpha serves as an activator in vitro and in vivo in humans. Treatment of normal monocytes or mononuclear cells in vitro with IFN-alpha caused a dose-dependent increase in monocyte NOS2 activity and NO production, and increased expression of NOS2 protein and mRNA expression. To determine if in vivo administration of IFN-alpha also modulated NOS2, we studied blood cells from patients with hepatitis C before and after IFN alpha therapy. Untreated patients with chronic hepatitis C virus infection had levels of NOS activity and NOS2 antigen in freshly isolated mononuclear cells similar to those of healthy subjects, and they expressed minimal or no NOS2 mRNA. However, IFN-alpha treatment of patients with hepatitis C infection was associated with a significant elevation in mononuclear cell NOS activity, NOS2 antigen content, and NOS2 mRNA content. IFN-alpha-treated patients had significant decreases in levels of serum alanine aminotransferase and plasma hepatitis C mRNA. The degree of IFN-alpha-enhanced mononuclear cell NOS2 antigen content correlated significantly with the degree of reduction in serum alanine aminotransferase levels. Thus, IFN-alpha treatment of cells in vitro or administration of IFN-alpha to hepatitis C patients in vivo increases expression of mononuclear cell NOS2 mRNA expression, NOS activity, NOS2 antigen expression, and NO production. Since NO has been reported to have antiviral activity for a variety of viruses, we speculate that induced NO production may be related to the antiviral action(s) of IFN-alpha in hepatitis C infection. PMID- 9348308 TI - Specific activation of the cysteine protease CPP32 during the negative selection of T cells in the thymus. AB - Cysteine proteases of the CED-3 and ICE family have been recently proposed as the ultimate executioners in several mammalian cell death pathways. Among them, the cysteine protease CPP32 has been shown to participate in programmed cell death (PCD), or apoptosis, affecting lymphoid cells in vitro. In the thymus, negative selection is a mechanism through which developing thymocytes expressing a TcR with high affinity for self peptide-MHC complexes are eliminated by PCD. In order to investigate the role of CPP32 in thymic apoptosis, isolated thymocytes were submitted to cell surface CD3 crosslinking by immobilized anti-CD3 mAb or to dexamethasone treatment. Although apoptosis occurred in the absence or after crosslinking with anti-CD3 mAb, specific activation of CPP32, as assessed by the extent of proteolytic cleavage of the p32 zymogen, was only detected in thymocytes cultured in the presence of the immobilized antibody or dexamethasone. This activation was a very early event during apoptosis as it occurred before the exposure of phosphatidyl serine to the upper side of the cell membrane. This was observed both in anti-CD3- and dexamethasone-induced apoptosis. Moreover, using mice transgenic for pigeon cytochrome C (PCC)-specific TcR, we were able to show that, after injection of PCC, the activation of CPP32 and cleavage of its substrate occurred in thymocytes obtained from mice expressing a permissive MHC haplotype for PCC presentation (H-2k). Moreover, PCC induced apoptosis was blocked by the caspase inhibitor zVAD. While spontaneous apoptosis was not accompanied by detectable levels of CPP32 processing, it was characterized by the proteolysis of poly(ADP-ribose) polymerase (PARP) and was blocked by the cysteine protease inhibitor, zVAD-CH2F. Taken together, these results support the concept that CPP32 is among the earliest effectors of the pathway leading to negative selection of autoreactive thymocytes. Our results also suggest the involvement of a distinct CPP32-like cysteine protease in spontaneous apoptosis of thymocytes. PMID- 9348309 TI - Enforced Bcl-2 expression inhibits antigen-mediated clonal elimination of peripheral B cells in an antigen dose-dependent manner and promotes receptor editing in autoreactive, immature B cells. AB - The mechanisms that establish immune tolerance in immature and mature B cells appear to be distinct. Membrane-bound autoantigen is thought to induce developmental arrest and receptor editing in immature B cells, whereas mature B cells have shortened lifespans when exposed to the same stimulus. In this study, we used Emu-bcl-2-22 transgenic (Tg) mice to test the prediction that enforced expression of the Bcl-2 apoptotic inhibitor in B cells would rescue mature, but not immature, B cells from tolerance induction. To monitor tolerance to the natural membrane autoantigen H-2Kb, we bred 3-83mudelta (anti-Kk,b) Ig Tg mice to H-2(b) mice or to mice expressing transgene-driven Kb in the periphery. In 3 83mudelta/bcl-2 Tg mice, deletion of autoreactive B cells induced by peripheral Kb antigen expression in the liver (MT-Kb Tg) or epithelia (KerIV-Kb Tg), was partly or completely inhibited, respectively. Furthermore, Bcl-2 protected peritoneal B-2 B cells from deletion mediated by acute antigen exposure, but this protection could be overcome by higher antigen dose. In contrast to its ability to block peripheral self-tolerance, Bcl-2 overexpression failed to inhibit central tolerance induced by bone marrow antigen expression, but instead, enhanced the receptor editing process. These studies indicate that apoptosis plays distinct roles in central and peripheral B cell tolerance. PMID- 9348310 TI - Interferon consensus sequence binding protein-deficient mice display impaired resistance to intracellular infection due to a primary defect in interleukin 12 p40 induction. AB - Mice lacking the transcription factor interferon consensus sequence binding protein (ICSBP), a member of the interferon regulatory factor family of transcription proteins, were infected with the intracellular protozoan, Toxoplasma gondii. ICSBP-deficient mice exhibited unchecked parasite replication in vivo and rapidly succumbed within 14 d after inoculation with an avirulent Toxoplasma strain. In contrast, few intracellular parasites were observed in wild type littermates and these animals survived for at least 60 d after infection. Analysis of cytokine synthesis in vitro and in vivo revealed a major deficiency in the expression of both interferon (IFN)-gamma and interleukin (IL)-12 p40 in the T. gondii exposed ICSBP-/- animals. In related experiments, macrophages from uninfected ICSBP-/- mice were shown to display a selective impairment in the mRNA expression of IL-12 p40 but not IL-1alpha, IL-1beta, IL-1Ra, IL-6, IL-10, or TNF alpha in response to live parasites, parasite antigen, lipopolysaccharide, or Staphylococcus aureus. This selective defect in IL-12 p40 production was observed regardless of whether the macrophages had been primed with IFN-gamma. We hypothesize that the impaired synthesis of IL-12 p40 in ICSBP-/- animals is the primary lesion responsible for the loss in resistance to T. gondii because IFN gamma-induced parasite killing was unimpaired in vitro and, more importantly, administration of exogenous IL-12 in vivo significantly prolonged survival of the infected mice. Together these findings implicate ICSBP as a major transcription factor which directly or indirectly regulates IL-12 p40 gene activation and, as a consequence, IFN-gamma-dependent host resistance. PMID- 9348311 TI - Interferon (IFN) consensus sequence-binding protein, a transcription factor of the IFN regulatory factor family, regulates immune responses in vivo through control of interleukin 12 expression. AB - Mice with a null mutation of the gene encoding interferon consensus sequence binding protein (ICSBP) develop a chronic myelogenous leukemia-like syndrome and mount impaired responses to certain viral and bacterial infections. To gain a mechanistic understanding of the contributions of ICSBP to humoral and cellular immunity, we characterized the responses of control and ICSBP-/- mice to infection with influenza A (flu) and Leishmania major (L. major). Mice of both genotypes survived infections with flu, but differed markedly in the isotype distribution of antiflu antibodies. In sera of normal mice, immunoglobulin (Ig)G2a antibodies were dominant over IgG1 antibodies, a pattern indicative of a T helper cell type 1 (Th1)-driven response. In sera of ICSBP-/- mice, however, IgG1 antibodies dominated over IgG2a antibodies, a pattern indicative of a Th2 driven response. The dominance of IgG1 and IgE over IgG2a was detected in the sera of uninfected mice as well. A seeming Th2 bias of ICSBP-deficient mice was also uncovered in their inability to control infection with L. major, where resistance is known to be dependent on IL-12 and IFN-gamma as components of a Th1 response. Infected ICSBP-deficient mice developed fulminant, disseminated leishmaniasis as a result of failure to mount a Th1-mediated curative response, although T cells remained capable of secreting IFN-gamma and macrophages of producing nitric oxide. Compromised Th1 differentiation in ICSBP-/- mice could not be attributed to hyporesponsiveness of CD4(+) T cells to interleukin (IL)-12; however, the ability of uninfected and infected ICSBP-deficient mice to produce IL-12 was markedly impaired. This indicates that ICSBP is a deciding factor in Th responses governing humoral and cellular immunity through its role in regulating IL-12 expression. PMID- 9348312 TI - Augmented expression of a human gene for 8-oxoguanine DNA glycosylase (MutM) in B lymphocytes of the dark zone in lymph node germinal centers. AB - B cells that mediate normal, T cell-dependent, humoral immune responses must first pass through germinal centers (GCs) within the cortex of antigenically stimulated lymph nodes. As they move through the dark zone and then the light zone in the GC, B cells are subjected to somatic hypermutation and switch recombination within their rearranged immunoglobulin genes and also participate in a number of other processes that control development into memory cells or cells specialized for antibody secretion. To investigate the molecular mechanisms that contribute to B cell development within GCs, we constructed a recombinant DNA library enriched for cDNAs derived from human genes expressed in B cells at this site. This library was found to contain a cDNA structurally and functionally related to genes in bacteria and yeast for the DNA repair enzyme 8-oxoguanine DNA glycosylase. Northern blot analysis indicated that the human gene is expressed as two alternatively spliced messenger RNAs within GC B cells at levels greatly exceeding that found in other tissues. In situ hybridization studies revealed that expression of this gene is most abundant within the dark zones of GCs. Both the function and localized expression of this gene suggest that it may play a role in somatic hypermutation of immunoglobulin genes. PMID- 9348314 TI - Association of glucocorticoid insensitivity with increased expression of glucocorticoid receptor beta. AB - In many chronic inflammatory disorders, glucocorticoid (GC) insensitivity is a challenging clinical problem associated with life-threatening disease progression. The molecular basis of GC insensitivity, however, is unknown. Alternative splicing of the GC receptor (R) pre-messenger RNA generates a second GCR, termed GCR-beta, which does not bind GCs but antagonizes the transactivating activity of the classic GCR, termed GCR-alpha. In the current study, we demonstrate that GC-insensitive asthma is associated with a significantly higher number of GCR-beta-immunoreactive cells in peripheral blood than GC-sensitive asthmatics or normal controls. Furthermore, we show that patients with GC insensitive asthma have cytokine-induced abnormalities in the DNA binding capability of the GCR. These abnormalities can be reproduced by transfection of cell lines with the GCR-beta gene resulting in significant reduction of their GCR alpha DNA binding capacity. We conclude that increased expression of GCR-beta is cytokine inducible and may account for GC insensitivity in this common inflammatory condition. PMID- 9348313 TI - Protection against invasive amebiasis by a single monoclonal antibody directed against a lipophosphoglycan antigen localized on the surface of Entamoeba histolytica. AB - A panel of monoclonal antibodies was raised from mice immunized with a membrane preparation from Entamoeba histolytica, the pathogenic species causing invasive amebiasis in humans. Antibody EH5 gave a polydisperse band in immunoblots from membrane preparations from different E. histolytica strains, and a much weaker signal from two strains of the nonpathogenic species Entamoeba dispar. Although the exact chemical structure of the EH5 antigen is not yet known, the ability of the antigen to be metabolically radiolabeled with [32P]phosphate or [3H]glucose, its sensitivity to digestion by mild acid and phosphatidylinositol-specific phospholipase C, and its specific extraction from E. histolytica trophozoites by a method used to prepare lipophosphoglycans from Leishmania showed that it could be classified as an amebal lipophosphoglycan. Confocal immunofluorescence and immunogold labeling of trophozoites localized the antigen on the outer face of the plasma membrane and on the inner face of internal vesicle membranes. Antibody EH5 strongly agglutinated amebas in a similar way to concanavalin A (Con A), and Con A bound to immunoaffinity-purified EH5 antigen. Therefore, surface lipophosphoglycans may play an important role in the preferential agglutination of pathogenic amebas by Con A. The protective ability of antibody EH5 was tested in a passive immunization experiment in a severe combined immunodeficient (SCID) mouse model. Intrahepatic challenge of animals after administration of an isotype matched control antibody or without treatment led to the development of a liver abscess in all cases, whereas 11 out of 12 animals immunized with the EH5 antibody developed no liver abscess. Our results demonstrate the importance and, for the first time, the protective capacity of glycan antigens on the surface of the amebas. PMID- 9348315 TI - Downregulated expression of SHP-1 in Burkitt lymphomas and germinal center B lymphocytes. AB - We wish to identify developmental changes in germinal center B cells that may contribute to their rapid growth. SHP-1 is an SH2 domain-containing phosphotyrosine phosphatase that negatively regulates activation of B cells and other cells of hematopoietic lineages. We have found that in all 13 EBV-negative and 11 EBV-positive Burkitt lymphomas with a nonlymphoblastoid phenotype, the mean concentration of SHP-1 was reduced to 5% of that of normal B and T cells. The possibility that this diminished expression of SHP-1 was related to the germinal center phenotype of Burkitt lymphomas was supported by the low to absent immunofluorescent staining for SHP-1 in germinal centers, and by the inverse relationship between the concentration of SHP-1 and the expression of the germinal center marker CD38 on purified tonsillar B cells. In CD38-high B cells, SHP-1 concentration was 20% of that of mantle zone B cells from the same donor. This reduction in SHP-1 is comparable to that of cells from motheaten viable mev/mev mice in which there is dysregulated, spontaneous signaling by cytokine and antigen receptors. Therefore, germinal center B cells may have a developmentally regulated, low threshold for cellular activation. PMID- 9348316 TI - Critical points of tumor necrosis factor action in central nervous system autoimmune inflammation defined by gene targeting. AB - Tumor necrosis factor (TNF)-dependent sites of action in the generation of autoimmune inflammation have been defined by targeted disruption of TNF in the C57BL/6 mouse strain. C57BL/6 mice are susceptible to an inflammatory, demyelinating form of experimental autoimmune encephalomyelitis (EAE) induced by the 35-55 peptide of myelin oligodendrocyte glycoprotein. Direct targeting of a strain in which EAE was inducible was necessary, as the location of the TNF gene renders segregation of the mutated allele from the original major histocompatibility complex by backcrossing virtually impossible. In this way a single gene effect was studied. We show here that TNF is obligatory for normal initiation of the neurological deficit, as demonstrated by a significant (6 d) delay in disease in its absence relative to wild-type (WT) mice. During this delay, comparable numbers of leukocytes were isolated from the perfused central nervous system (CNS) of WT and TNF-/- mice. However, in the TNF-/- mice, immunohistological analysis of CNS tissue indicated that leukocytes failed to form the typical mature perivascular cuffs observed in WT mice at this same time point. Severe EAE, including paralysis and widespread CNS perivascular inflammation, eventually developed without TNF. TNF-/- and WT mice recovered from the acute illness at the same time, such that the overall disease course in TNF-/ mice was only 60% of the course in control mice. Primary demyelination occurred in both WT and TNF-/- mice, although it was of variable magnitude. These results are consistent with the TNF dependence of processes controlling initial leukocyte movement within the CNS. Nevertheless, potent alternative mechanisms exist to mediate all other phases of EAE. PMID- 9348317 TI - Antiviral activity of tumor necrosis factor (TNF) is mediated via p55 and p75 TNF receptors. AB - The antiviral nature of tumor necrosis factor (TNF) is generally well accepted. TNF appears to induce multiple antiviral mechanisms, and to synergize with interferon (IFN)-gamma in promoting antiviral activities. We infected TNF receptor (TNFR)-deficient mice with the virulent murine pathogen, ectromelia virus (EV), and observed that otherwise resistant mice were susceptible to lethal infection. To study the molecular basis of the antiviral action of TNF, mice were infected with a recombinant vaccinia virus encoding murine TNF (VV-HA-TNF). In normal mice, the replication of VV-HA-TNF was highly attenuated. In contrast, mice in which the TNFR type 1 (p55) or the TNFR type 2 (p75) were genetically disrupted showed a moderate defect in their capacity to clear the TNF-encoding virus. The contribution of both TNF receptors to the control of VV-HA-TNF was confirmed by the enhanced replication of VV-HA-TNF in mice deficient for both p55 and p75. These observations were corroborated by infecting TNFR-deficient mice with EV. For both infections, the p55 and p75 TNFRs were necessary to maintain normal levels of resistance. Thus, the antiviral activity of TNF is mediated via both TNFRs in vivo. Furthermore, these studies establish that TNF is an important component of the host response to a natural virus infection. PMID- 9348318 TI - Inhibition of T cell and promotion of natural killer cell development by the dominant negative helix loop helix factor Id3. AB - Bipotential T/natural killer (NK) progenitor cells are present in the human thymus. Despite their bipotential capacity, these progenitors develop predominantly to T cells in the thymus. The mechanisms controlling this developmental choice are unknown. Here we present evidence that a member(s) of the family of basic helix loop helix (bHLH) transcription factors determines lineage specification of NK/T cell progenitors. The natural dominant negative HLH factor Id3, which blocks transcriptional activity of a number of known bHLH factors, was expressed in CD34+ progenitor cells by retrovirus-mediated gene transfer. Constitutive expression of Id3 completely blocks development of CD34+ cells into T cells in a fetal thymic organ culture (FTOC). In contrast, development into NK cells in an FTOC is enhanced. Thus, the activity of a bHLH transcription factor is necessary for T lineage differentiation of bipotential precursors, in the absence of which a default pathway leading to NK cell development is chosen. Our results identify a molecular switch for lineage specification in early lymphoid precursors of humans. PMID- 9348319 TI - Prostaglandin E2 and tumor necrosis factor alpha cooperate to activate human dendritic cells: synergistic activation of interleukin 12 production. AB - Interleukin (IL)-12 is a proinflammatory cytokine that contributes to innate resistance and to the development of antigen-specific T cell responses. Among other effects, prostaglandin E2 (PGE2) inhibits the production of IL-12 by macrophages activated with lipopolysaccharide (LPS). Here we investigated the effects of PGE2 on human dendritic cells (DCs) which develop in the presence of GM-CSF and IL-4. We demonstrate that in the absence of LPS, PGE2 dose dependently stimulated the production of IL-12 by DCs. Although PGE2 alone stimulated the production of low amounts of IL-12 only, it synergized with tumor necrosis factor (TNF)-alpha to induce high levels of IL-12 production by DCs. Addition of TNF alpha in the absence of PGE2 had no effect on IL-12 production. Conversely, in the presence of LPS, PGE2 inhibited IL-12 production by DCs in a dose-dependent manner. The combination of PGE2 and TNF-alpha efficiently silenced mannose receptor-mediated endocytosis in DCs and readily induced neo-expression of the CD83 antigen. In addition, the expression of various surface antigens such as major histocompatibility complex class I and II, adhesion, as well as costimulatory molecules was upregulated by this treatment. The effects of PGE2 on IL-12 synthesis and CD83 expression could be mimicked by dibutyryl-cAMP and forskolin, indicating that they were due to the intracellular elevation of cAMP levels. DC treated with PGE2 and TNF-alpha were most potent in stimulating allogeneic T cell proliferation. Our data demonstrate that PGE2 contributes to the maturation of human DCs and that PGE2 can be a potent enhancer of IL-12 production by human DCs. PMID- 9348320 TI - Generation of lytic natural killer 1.1+, Ly-49- cells from multipotential murine bone marrow progenitors in a stroma-free culture: definition of cytokine requirements and developmental intermediates. AB - We have developed a stroma-free culture system in which mouse marrow or thymus cells, known to be enriched for lymphoid progenitors, can be driven to generate natural killer (NK) cells. Culture of lineage marker (Lin)-, c-kit+, Sca2+, interleukin (IL)-2/15Rbeta (CD122)- marrow cells in IL-6, IL-7, stem cell factor (SCF), and flt3 ligand (flt3-L) for 5-6 d followed by IL-15 alone for an additional 4-5 d expanded the starting population 30-40-fold and gave rise to a virtually pure population of NK1.1+, CD3- cells. Preculture in IL-6, IL-7, SCF, and flt3-L was necessary for inducing IL-15 responsiveness in the progenitors because the cells failed to significantly expand when cultured in IL-15 alone from the outset. Although culture of the sorted progenitors in IL-6, IL-7, SCF, and flt3-L for the entire 9-11-d culture period caused significant expansion, no lytic NK1.1+ cells were generated if IL-15 was not added, demonstrating a critical role for IL-15 in NK differentiation. Thus, two distinct populations of NK progenitors, IL-15 unresponsive and IL-15 responsive, have been defined. Similar results were obtained with Lin-, CD44+, CD25-, c-kit+ lymphoid progenitors obtained from adult thymus. The NK cells generated by this protocol lysed the NK-sensitive target YAC-1 and expressed markers of mature NK cells with the notable absence of Ly-49 major histocompatibility complex (MHC) receptors. However, despite the apparent lack of these inhibitory MHC receptors, the NK cells generated could distinguish MHC class I+ from class I- syngeneic targets, suggesting the existence of novel class I receptors. PMID- 9348321 TI - Wound healing is accelerated by agonists of adenosine A2 (G alpha s-linked) receptors. AB - The complete healing of wounds is the final step in a highly regulated response to injury. Although many of the molecular mediators and cellular events of healing are known, their manipulation for the enhancement and acceleration of wound closure has not proven practical as yet. We and others have established that adenosine is a potent regulator of the inflammatory response, which is a component of wound healing. We now report that ligation of the G alpha s-linked adenosine receptors on the cells of an artificial wound dramatically alters the kinetics of wound closure. Excisional wound closure in normal, healthy mice was significantly accelerated by topical application of the specific A2A receptor agonist CGS-21680 (50% closure by day 2 in A2 receptor antagonists. In rats rendered diabetic (streptozotocin-induced diabetes mellitus) wound healing was impaired as compared to nondiabetic rats; CGS-21680 significantly increased the rate of wound healing in both nondiabetic and diabetic rats. Indeed, the rate of wound healing in the CGS-21680-treated diabetic rats was greater than or equal to that observed in untreated normal rats. These results appear to constitute the first evidence that a small molecule, such as an adenosine receptor agonist, accelerates wound healing in both normal animals and in animals with impaired wound healing. PMID- 9348322 TI - Stoichiometry of recombinant N-methyl-D-aspartate receptor channels inferred from single-channel current patterns. AB - Single-channel currents were recorded from mouse NR1-NR2B (zeta-epsilon2) receptors containing mixtures of wild-type and mutant subunits expressed in Xenopus oocytes. Mutant subunits had an asparagine-to-glutamine (N-to-Q) mutation at the N0 site of the M2 segment (NR1:598, NR2B:589). Receptors with pure N or Q NR1 and NR2 subunits generated single-channel currents with distinctive current patterns. Based on main and sublevel amplitudes, occupancy probabilities, and lifetimes, four patterns of current were identified, corresponding to receptors with the following subunit compositions (NR1/NR2): N/N, N/Q, Q/N, and Q/Q. Only one current pattern was apparent for each composition. When a mixture of N and Q NR2 subunits was coexpressed with pure mutant NR1 subunits, three single-channel current patterns were apparent. One pattern was the same as Q/Q receptors and another was the same as Q/N receptors. The third, novel pattern presumably arose from hybrid receptors having both N and Q NR2 subunits. When a mixture of N and Q NR1 subunits was coexpressed with pure mutant NR2 subunits, six single-channel current patterns were apparent. One pattern was the same as Q/Q receptors and another was the same as N/Q receptors. The four novel patterns presumably arose from hybrid receptors having both N and Q NR1 subunits. The relative frequency of NR1 hybrid receptor current patterns depended on the relative amounts of Q and N subunits that were injected into the oocytes. The number of hybrid receptor patterns suggests that there are two NR2 subunits per receptor and is consistent with either three or five NR1 subunits per receptor, depending on whether or not the order of mutant and wild-type subunits influences the current pattern. When considered in relation to other studies, the most straightforward interpretation of the results is that N-methyl-D-aspartate receptors are pentamers composed of three NR1 and two NR2 subunits. PMID- 9348323 TI - Intracellular Ca2+ inhibits smooth muscle L-type Ca2+ channels by activation of protein phosphatase type 2B and by direct interaction with the channel. AB - Modulation of L-type Ca2+ channels by tonic elevation of cytoplasmic Ca2+ was investigated in intact cells and inside-out patches from human umbilical vein smooth muscle. Ba2+ was used as charge carrier, and run down of Ca2+ channel activity in inside-out patches was prevented with calpastatin plus ATP. Increasing cytoplasmic Ca2+ in intact cells by elevation of extracellular Ca2+ in the presence of the ionophore A23187 inhibited the activity of L-type Ca2+ channels in cell-attached patches. Measurement of the actual level of intracellular free Ca2+ with fura-2 revealed a 50% inhibitory concentration (IC50) of 260 nM and a Hill coefficient close to 4 for Ca2+- dependent inhibition. Ca2+-induced inhibition of Ca2+ channel activity in intact cells was due to a reduction of channel open probability and availability. Ca2+-induced inhibition was not affected by the protein kinase inhibitor H-7 (10 microM) or the cytoskeleton disruptive agent cytochalasin B (20 microM), but prevented by cyclosporin A (1 microg/ ml), an inhibitor of protein phosphatase 2B (calcineurin). Elevation of Ca2+ at the cytoplasmic side of inside-out patches inhibited Ca2+ channels with an IC50 of 2 microM and a Hill coefficient close to unity. Direct Ca2+-dependent inhibition in cell-free patches was due to a reduction of open probability, whereas availability was barely affected. Application of purified protein phosphatase 2B (12 U/ml) to the cytoplasmic side of inside-out patches at a free Ca2+ concentration of 1 microM inhibited Ca2+ channel open probability and availability. Elevation of cytoplasmic Ca2+ in the presence of PP2B, suppressed channel activity in inside-out patches with an IC50 of approximately 380 nM and a Hill coefficient of approximately 3; i.e., characteristics reminiscent of the Ca2+ sensitivity of Ca2+ channels in intact cells. Our results suggest that L-type Ca2+ channels of smooth muscle are controlled by two Ca2+-dependent negative feedback mechanisms. These mechanisms are based on (a) a protein phosphatase 2B-mediated dephosphorylation process, and (b) the interaction of intracellular Ca2+ with a single membrane-associated site that may reside on the channel protein itself. PMID- 9348324 TI - Calcium modulation of ligand affinity in the cyclic GMP-gated ion channels of cone photoreceptors. AB - To investigate modulation of the activation of cGMP-gated ion channels in cone photoreceptors, we measured currents in membrane patches detached from the outer segments of single cones isolated from striped bass retina. The sensitivity of these channels to activation by cGMP depends on the history of exposure to divalent cations of the membrane's cytoplasmic surface. In patches maintained in 20 microM Ca++ and 100 microM Mg++ after excision, the current amplitude dependence on cGMP is well described by a Hill equation with average values of K1/2, the concentration necessary to activate half the maximal current, of 86 microM and a cooperativity index, n, of 2.57. Exposing the patch to a solution free of divalent cations irreversibly increases the cGMP sensitivity; the average value of K1/2 shifts to 58.8 microM and n shifts to 1.8. Changes in cGMP sensitivity do not affect other functional parameters of the ion channels, such as the interaction and permeation of mono- and divalent cations. Modulation of cGMP activation depends on the action of an endogenous factor that progressively dissociates from the channel as Ca++ concentration is lowered below 1 microM. The activity of the endogenous modulator is not well mimicked by exogenously added calmodulin, although this protein competes with the endogenous modulator for a common binding site. Thus, the modulation of cGMP affinity in cones depends on the activity of an unidentified molecule that may not be calmodulin. PMID- 9348325 TI - Inositol 1,4,5-trisphosphate (InsP3) and calcium interact to increase the dynamic range of InsP3 receptor-dependent calcium signaling. AB - The inositol 1,4,5-trisphosphate (InsP3)-gated Ca channel in cerebellum is tightly regulated by Ca (Bezprozvanny, I., J. Watras, and B.E. Ehrlich. 1991. Nature (Lond.). 351:751-754; Finch, E.A., T. J. Turner, and S.M. Goldin. 1991. Science (Wash. DC). 252:443-446; Hannaert-Merah, Z., J.F. Coquil, L. Combettes, M. Claret, J.P. Mauger, and P. Champeil. 1994. J. Biol. Chem. 269:29642-29649; Iino, M. 1990. J. Gen. Physiol. 95:1103-1122; Marshall, I., and C. Taylor. 1994. Biochem. J. 301:591-598). In previous single channel studies, the Ca dependence of channel activity, monitored at 2 microM InsP3, was described by a bell-shaped curve (Bezprozvanny, I., J. Watras, and B.E. Ehrlich. 1991. Nature (Lond.). 351:751-754). We report here that, when we used lower InsP3 concentrations, the peak of the Ca-dependence curve shifted to lower Ca concentrations. Unexpectedly, when we used high InsP3 concentrations, channel activity persisted at Ca concentrations as high as 30 microM. To explore this unexpected response of the channel, we measured InsP3 binding over a broad range of InsP3 concentrations. We found the well-characterized high affinity InsP3 binding sites (with Kd < 1 and 50 nM) (Maeda, N., M. Niinobe, and K. Mikoshiba. 1990. EMBO (Eur. Mol. Biol. Organ.) J. 9:61-67; Mignery, G., T.C. Sudhof, K. Takei, and P. De Camilli. 1989. Nature (Lond.). 342:192-195; Ross, C.A., J. Meldolesi, T.A. Milner, T. Satoh, S. Supattapone, and S.H. Snyder. 1989. Nature (Lond.). 339:468-470) and a low affinity InsP3 binding site (Kd = 10 microM). Using these InsP3 binding sites, we developed a new model that accounts for the shift in the Ca-dependence curve at low InsP3 levels and the maintained channel activity at high Ca and InsP3 levels. The observed Ca dependence of the InsP3-gated Ca channel allows the cell to abbreviate the rise of intracellular Ca in the presence of low levels of InsP3, but also provides a means of maintaining high intracellular Ca during periods of prolonged stimulation. PMID- 9348326 TI - Ion conduction through C-type inactivated Shaker channels. AB - C-type inactivation of Shaker potassium channels involves entry into a state (or states) in which the inactivated channels appear nonconducting in physiological solutions. However, when Shaker channels, from which fast N-type inactivation has been removed by NH2-terminal deletions, are expressed in Xenopus oocytes and evaluated in inside-out patches, complete removal of K+ ions from the internal solution exposes conduction of Na+ and Li+ in C-type inactivated conformational states. The present paper uses this observation to investigate the properties of ion conduction through C-type inactivated channel states, and demonstrates that both activation and deactivation can occur in C-type states, although with slower than normal kinetics. Channels in the C-type states appear "inactivated" (i.e., nonconducting) in physiological solutions due to the summation of two separate effects: first, internal K+ ions prevent Na+ ions from permeating through the channel; second, C-type inactivation greatly reduces the permeability of K+ relative to the permeability of Na+, thus altering the ion selectivity of the channel. PMID- 9348327 TI - Mechanism of ion permeation in skeletal muscle chloride channels. AB - Voltage-gated Cl- channels belonging to the ClC family exhibit unique properties of ion permeation and gating. We functionally probed the conduction pathway of a recombinant human skeletal muscle Cl- channel (hClC-1) expressed both in Xenopus oocytes and in a mammalian cell line by investigating block by extracellular or intracellular I- and related anions. Extracellular and intracellular I- exert blocking actions on hClC-1 currents that are both concentration and voltage dependent. Similar actions were observed for a variety of other halide (Br-) and polyatomic (SCN-, NO3-, CH3SO3-) anions. In addition, I- block is accompanied by gating alterations that differ depending on which side of the membrane the blocker is applied. External I- causes a shift in the voltage-dependent probability that channels exist in three definable kinetic states (fast deactivating, slow deactivating, nondeactivating), while internal I- slows deactivation. These different effects on gating properties can be used to distinguish two functional ion binding sites within the hClC-1 pore. We determined KD values for I- block in three distinct kinetic states and found that binding of I- to hClC-1 is modulated by the gating state of the channel. Furthermore, estimates of electrical distance for I- binding suggest that conformational changes affecting the two ion binding sites occur during gating transitions. These results have implications for understanding mechanisms of ion selectivity in hClC-1, and for defining the intimate relationship between gating and permeation in ClC channels. PMID- 9348328 TI - Sodium and calcium inward currents in freshly dissociated smooth myocytes of rat uterus. AB - Freshly dissociated myocytes from nonpregnant, pregnant, and postpartum rat uteri have been studied with the tight-seal patch-clamp method. The inward current contains both INa and ICa that are vastly different from those in tissue-cultured material. INa is abolished by Na+-free medium and by 1 microM tetrodotoxin. It first appears at approximately -40 mV, reaches maximum at 0 mV, and reverses at 84 mV. It activates with a voltage-dependent tau of 0.2 ms at 20 mV, and inactivates as a single exponential with a tau of 0. 4 ms. Na+ conductance is half activated at -21.5 mV, and half inactivated at -59 mV. INa reactivates with a tau of 20 ms. ICa is abolished by Ca2+-free medium, Co2+ (5 mM), or nisoldipine (2 microM), and enhanced in 30 mM Ca2+, Ba2+, or BAY-K 8644. It first appears at approximately -30 mV and reaches maximum at +10 mV. It activates with a voltage dependent tau of 1.5 ms at 20 mV, and inactivates in two exponential phases, with tau's of 33 and 133 ms. Ca2+ conductance is half activated at -7.4 mV, and half inactivated at -34 mV. ICa reactivates with tau's of 27 and 374 ms. INa and ICa are seen in myocytes from nonpregnant estrus uteri and throughout pregnancy, exhibiting complex changes. The ratio of densities of peak INa/ICa changes from 0.5 in the nonpregnant state to 1.6 at term. The enhanced role of INa, with faster kinetics, allows more frequent repetitive spike discharges to facilitate simultaneous excitation of the parturient uterus. In postpartum, both currents decrease markedly, with INa vanishing from most myocytes. Estrogen-enhanced genomic influences may account for the emergence of INa, and increased densities of INa and ICa as pregnancy progresses. Other influences may regulate varied channel expression at different stages of pregnancy. PMID- 9348329 TI - Correlation between charge movement and ionic current during slow inactivation in Shaker K+ channels. AB - Prolonged depolarization induces a slow inactivation process in some K+ channels. We have studied ionic and gating currents during long depolarizations in the mutant Shaker H4-Delta(6-46) K+ channel and in the nonconducting mutant (Shaker H4-Delta(6-46)-W434F). These channels lack the amino terminus that confers the fast (N-type) inactivation (Hoshi, T., W.N. Zagotta, and R.W. Aldrich. 1991. Neuron. 7:547-556). Channels were expressed in oocytes and currents were measured with the cut-open-oocyte and patch-clamp techniques. In both clones, the curves describing the voltage dependence of the charge movement were shifted toward more negative potentials when the holding potential was maintained at depolarized potentials. The evidences that this new voltage dependence of the charge movement in the depolarized condition is associated with the process of slow inactivation are the following: (a) the installation of both the slow inactivation of the ionic current and the inactivation of the charge in response to a sustained 1-min depolarization to 0 mV followed the same time course; and (b) the recovery from inactivation of both ionic and gating currents (induced by repolarizations to -90 mV after a 1-min inactivating pulse at 0 mV) also followed a similar time course. Although prolonged depolarizations induce inactivation of the majority of the channels, a small fraction remains non-slow inactivated. The voltage dependence of this fraction of channels remained unaltered, suggesting that their activation pathway was unmodified by prolonged depolarization. The data could be fitted to a sequential model for Shaker K+ channels (Bezanilla, F., E. Perozo, and E. Stefani. 1994. Biophys. J. 66:1011-1021), with the addition of a series of parallel nonconducting (inactivated) states that become populated during prolonged depolarization. The data suggest that prolonged depolarization modifies the conformation of the voltage sensor and that this change can be associated with the process of slow inactivation. PMID- 9348330 TI - Tetracaine reports a conformational change in the pore of cyclic nucleotide-gated channels. AB - Local anesthetics are a diverse group of clinically useful compounds that act as pore blockers of both voltage- and cyclic nucleotide-gated (CNG) ion channels. We used the local anesthetic tetracaine to probe the nature of the conformational change that occurs in the pore of CNG channels during the opening allosteric transition. When applied to the intracellular side of wild-type rod CNG channels expressed in Xenopus oocytes from the alpha subunit, the local anesthetic tetracaine exhibits state-dependent block, binding with much higher affinity to closed states than to open states. Here we show that neutralization of a glutamic acid in the conserved P region (E363G) eliminated this state dependence of tetracaine block. Tetracaine blocked E363G channels with the same effectiveness at high concentrations of cGMP, when the channel spent more time open, and at low concentrations of cGMP, when the channel spent more time closed. In addition, Ni2+, which promotes the opening allosteric transition, decreased the effectiveness of tetracaine block of wild-type but not E363G channels. Similar results were obtained in a chimeric CNG channel that exhibits a more favorable opening allosteric transition. These results suggest that E363 is accessible to internal tetracaine in the closed but not the open configuration of the pore and that the conformational change that accompanies channel opening includes a change in the conformation or accessibility of E363. PMID- 9348331 TI - Modulation of the Kv1.3 potassium channel by receptor tyrosine kinases. AB - The voltage-dependent potassium channel, Kv1.3, is modulated by the epidermal growth factor receptor (EGFr) and the insulin receptor tyrosine kinases. When the EGFr and Kv1.3 are coexpressed in HEK 293 cells, acute treatment of the cells with EGF during a patch recording can suppress the Kv1.3 current within tens of minutes. This effect appears to be due to tyrosine phosphorylation of the channel, as it is blocked by treatment with the tyrosine kinase inhibitor erbstatin, or by mutation of the tyrosine at channel amino acid position 479 to phenylalanine. Previous work has shown that there is a large increase in the tyrosine phosphorylation of Kv1.3 when it is coexpressed with the EGFr. Pretreatment of EGFr and Kv1.3 cotransfected cells with EGF before patch recording also results in a decrease in peak Kv1.3 current. Furthermore, pretreatment of cotransfected cells with an antibody to the EGFr ligand binding domain (alpha-EGFr), which blocks receptor dimerization and tyrosine kinase activation, blocks the EGFr-mediated suppression of Kv1.3 current. Insulin treatment during patch recording also causes an inhibition of Kv1.3 current after tens of minutes, while pretreatment for 18 h produces almost total suppression of current. In addition to depressing peak Kv1.3 current, EGF treatment produces a speeding of C-type inactivation, while pretreatment with the alpha-EGFr slows C type inactivation. In contrast, insulin does not influence C-type inactivation kinetics. Mutational analysis indicates that the EGF-induced modulation of the inactivation rate occurs by a mechanism different from that of the EGF-induced decrease in peak current. Thus, receptor tyrosine kinases differentially modulate the current magnitude and kinetics of a voltage-dependent potassium channel. PMID- 9348332 TI - Phe-Met-Arg-Phe-amide activates a novel voltage-dependent K+ current through a lipoxygenase pathway in molluscan neurones. AB - The neuropeptide Phe-Met-Arg-Phe-amide (FMRFa) dose dependently (ED50 = 23 nM) activated a K+ current in the peptidergic caudodorsal neurones that regulate egg laying in the mollusc Lymnaea stagnalis. Under standard conditions ([K+]o = 1.7 mM), only outward current responses occurred. In high K+ salines ([K+]o = 20 or 57 mM), current reversal occurred close to the theoretical reversal potential for K+. In both salines, no responses were measured below -120 mV. Between -120 mV and the K+ reversal potential, currents were inward with maximal amplitudes at approximately -60 mV. Thus, U-shaped current-voltage relations were obtained, implying that the response is voltage dependent. The conductance depended both on membrane potential and extracellular K+ concentration. The voltage sensitivity was characterized by an e-fold change in conductance per approximately 14 mV at all [K+]o. Since this result was also obtained in nearly symmetrical K+ conditions, it is concluded that channel gating is voltage dependent. In addition, outward rectification occurs in asymmetric K+ concentrations. Onset kinetics of the response were slow (rise time approximately 650 ms at -40 mV). However, when FMRFa was applied while holding the cell at -120 mV, to prevent activation of the current but allow activation of the signal transduction pathway, a subsequent step to -40 mV revealed a much more rapid current onset. Thus, onset kinetics are largely determined by steps preceding channel activation. With FMRFa applied at -120 mV, the time constant of activation during the subsequent test pulse decreased from approximately 36 ms at -60 mV to approximately 13 ms at -30 mV, confirming that channel opening is voltage dependent. The current inactivated voltage dependently. The rate and degree of inactivation progressively increased from -120 to -50 mV. The current is blocked by internal tetraethylammonium and by bath- applied 4-aminopyridine, tetraethylammonium, Ba2+, and, partially, Cd2+ and Cs+. The response to FMRFa was affected by intracellular GTPgammaS. The response was inhibited by blockers of phospholipase A2 and lipoxygenases, but not by a cyclo-oxygenase blocker. Bath applied arachidonic acid induced a slow outward current and occluded the response to FMRFa. These results suggest that the FMRFa receptor couples via a G-protein to the lipoxygenase pathway of arachidonic acid metabolism. The biophysical and pharmacological properties of this transmitter operated, but voltage-dependent K+ current distinguish it from other receptor-driven K+ currents such as the S current- and G-protein-dependent inward rectifiers. PMID- 9348333 TI - Intracellular Cl- dependence of Na-H exchange in barnacle muscle fibers under normotonic and hypertonic conditions. AB - We previously showed that shrinking a barnacle muscle fiber (BMF) in a hypertonic solution (1,600 mosM/kg) stimulates an amiloride-sensitive Na-H exchanger. This activation is mediated by a G protein and requires intracellular Cl-. The purpose of the present study was to determine (a) whether Cl- plays a role in the activation of Na-H exchange under normotonic conditions (975 mosM/kg), (b) the dose dependence of [Cl-]i for activation of the exchanger under both normo- and hypertonic conditions, and (c) the relative order of the Cl-- and G-protein dependent steps. We acid loaded BMFs by internally dialyzing them with a pH-6.5 dialysis fluid containing no Na+ and 0-194 mM Cl-. The artificial seawater bathing the BMF initially contained no Na+. After dialysis was halted, adding 50 mM Na+ to the artificial seawater caused an amiloride-sensitive pHi increase under both normo- and hypertonic conditions. The computed Na-H exchange flux (JNa H) increased with increasing [Cl-]i under both normo- and hypertonic conditions, with similar apparent Km values ( approximately 120 mM). However, the maximal JNa H increased by nearly 90% under hypertonic conditions. Thus, activation of Na-H exchange at low pHi requires Cl- under both normo- and hypertonic conditions, but at any given [Cl-]i, JNa-H is greater under hyper- than normotonic conditions. We conclude that an increase in [Cl-]i is not the primary shrinkage signal, but may act as an auxiliary shrinkage signal. To determine whether the Cl--dependent step is after the G-protein-dependent step, we predialyzed BMFs to a Cl--free state, and then attempted to stimulate Na-H exchange by activating a G protein. We found that, even in the absence of Cl-, dialyzing with GTPgammaS or AlF3, or injecting cholera toxin, stimulates Na-H exchange. Because Na-H exchange activity was absent in control Cl--depleted fibers, the Cl--dependent step is at or before the G protein in the shrinkage signal-transduction pathway. The stimulation by AlF3 indicates that the G protein is a heterotrimeric G protein. PMID- 9348334 TI - Cloning and characterization of murine glial cell-derived neurotrophic factor inducible transcription factor (MGIF). AB - The potent neurotrophic factor glial cell-derived neurotrophic factor (GDNF) is a distant member of the transforming growth factor-beta (TGF-beta) superfamily of proteins. We report a transcription factor that is the first nuclear protein known to be induced by GDNF, thus designated murine GDNF inducible factor (mGIF). The cDNA was cloned in the course of investigating transcription factors that bind to Sp1 consensus sequences, using the in situ filter detection method, and it was found to encode a protein having the same C2-H2 zinc finger motif as Sp1. Sequence analysis indicated that mGIF is homologous to the human TGF-beta inducible early gene (TIEG) and human early growth response gene-alpha (EGR alpha). mGIF is widely distributed in the adult mouse with high mRNA levels in kidney, lung, brain, liver, heart, and testis. In the adult brain, mGIF is abundantly expressed in hippocampus, cerebral cortex, cerebellum, and amygdala with lower amounts in striatum, nucleus accumbens, olfactory tubercle, thalamus, and substantia nigra. During development, mGIF mRNA also has a wide distribution, including in cerebral cortex, cerebellar primordium, kidney, intestine, liver, and lung. GDNF induces the expression of mGIF rapidly and transiently both in a neuroblastoma cell line and in primary cultures of rat embryonic cortical neurons. Co-transfection of the Drosophila SL2 cells using mGIF expression plasmid and reporter constructs having Sp1 binding sites indicated that mGIF represses transcription from a TATA-containing as well as from a TATA-less promoter. These observations suggest that the zinc finger transcription factor mGIF could be important in mediating some of the biological effects of GDNF. PMID- 9348335 TI - Brain-derived neurotrophic factor, neurotrophin-3, and neurotrophin-4 complement and cooperate with each other sequentially during visceral neuron development. AB - The neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and neurotrophin-4 (NT4) are crucial target-derived factors controlling the survival of peripheral sensory neurons during the embryonic period of programmed cell death. Recently, NT3 has also been found to act in a local manner on somatic sensory precursor cells during early development in vivo. Culture studies suggest that these cells switch dependency to NGF at later stages. The neurotrophins acting on the developing placode-derived visceral nodose/petrosal (N/P) ganglion neurons are BDNF, NT3, and NT4. To assess their roles in development, we analyzed embryonic development in mice carrying a deletion in each of these genes, or combinations of them, and found that they are essential in preventing the death of N/P ganglion neurons during different periods of embryogenesis. Both NT3 and NT4 are crucial during the period of ganglion formation, whereas BDNF acts later in development. Many, but not all, of the NT3- and NT4-dependent neurons switch to BDNF at later stages. We conclude that most of the N/P ganglion neurons depend on more than one neurotrophin and that they act in a complementary as well as a collaborative manner in a developmental sequence for the establishment of a full complement of visceral neurons. PMID- 9348337 TI - The mechanism of cAMP-mediated enhancement at a cerebellar synapse. AB - Increases in cAMP have been shown previously to enhance the strength of the granule cell to Purkinje cell synapse. We have examined the mechanisms underlying this enhancement in rat cerebellar brain slices. Elevation of cAMP levels by forskolin increased synaptic currents in a dose-dependent manner. Fluorometric calcium measurements revealed that forskolin did not affect presynaptic calcium influx or resting calcium levels. The waveform of the presynaptic volley was also unaltered, indicating that changes in the presynaptic action potential did not contribute to synaptic enhancement. However, forskolin enhanced the frequency but not the size of spontaneous miniature EPSCs. There was a one-to-one correspondence between increases of spontaneous and evoked neurotransmitter release. These results suggest that forskolin increases release at this synapse via presynaptic mechanisms that do not alter calcium influx. The effect of forskolin on paired-pulse facilitation was examined to assess the relative contributions of changes in the probability of release (p) and changes in the number of functional release sites (n) to this form of enhancement. These experiments suggest that although small changes in n cannot be excluded, most of the enhancement arises from increases in p. PMID- 9348336 TI - Ca2+ influx amplifies protein kinase C potentiation of recombinant NMDA receptors. AB - Protein kinase C (PKC) potentiates NMDA receptors in hippocampal, trigeminal, and spinal neurons. Although PKC phosphorylates the NMDA receptor subunit NR1 at four residues within the C terminal splice cassette C1, the molecular mechanisms underlying PKC potentiation of NMDA responses are not yet known. The present study examined the role of Ca2+ in PKC potentiation of recombinant NMDA receptors expressed in Xenopus oocytes. We found that Ca2+ influx through PKC-potentiated NMDA receptors can further increase the NMDA response ("Ca2+ amplification"). Ca2+ amplification required a rise in intracellular Ca2+ concentration at or near the intracellular end of the channel and was independent of Ca2+-activated Cl- current. Ca2+ amplification depended on extracellular Ca2+ concentration during NMDA application and not during PKC activation. Ca2+ amplification was reduced by the membrane-permeant Ca2+-chelating agent BAPTA-AM. Mutant receptors with greatly reduced Ca2+ permeability did not exhibit Ca2+ amplification. Receptors containing the NR1 N-terminal splice cassette showed more Ca2+ amplification, possibly because of their larger basal current and therefore greater Ca2+ influx. Contrary to expectation, splicing out the two C-terminal splice cassettes of NR1 enhanced PKC potentiation in a manner independent of extracellular Ca2+. This observation indicates that PKC potentiation does not require phosphorylation of the C1 cassette of the NR1 subunit. PKC potentiation of NMDA receptors in vivo is likely to be affected by Ca2+ amplification of the potentiated signal; the degree of amplification will depend in part on alternative splicing of the NR1 subunit, which is regulated developmentally and in a cell-specific manner. PMID- 9348338 TI - Peroxide modulation of slow onset potentiation in rat hippocampus. AB - Exposure of rat hippocampal slices to low concentrations of the muscarinic agonist carbachol (CCh) has been shown to produce a slow onset long-term potentiation (LTP) of reactivity to afferent stimulation in CA1 neurons. Although this potentiation shares a number of properties with tetanic LTP, muscarinic LTP (LTPm) is independent of activation of the NMDA receptor. We now demonstrate that low levels of hydrogen peroxide (H2O2) cause hippocampal slices to lose the ability to express LTPm. This powerful effect of H2O2 is selective in that it does not affect the reactivity of hippocampal neurons to higher concentrations of CCh. In fact, H2O2 also blocks induction of a slow onset, non-NMDA-dependent tetanic LTP (NN-LTP). The functional relevance of this action of H2O2 is exemplified by the fact that the hippocampus of aged rats, which produces higher levels of endogenous H2O2 than that of young rats, lacks LTPm and expresses a markedly reduced NN-LTP. In aged rats, the lack of LTPm contrasts with an apparently normal muscarinic suppression of the EPSP slope induced by higher concentrations of CCh. When hippocampal slices from aged animals are treated with catalase, an enzyme that breaks down H2O2, LTPm is restored, and NN-LTP is enhanced. Thus, our study proposes a unique and novel age-dependent peroxide regulation of LTPm in the brain and provides a link between the cholinergic system, aging, and memory functions. PMID- 9348339 TI - The murine P84 neural adhesion molecule is SHPS-1, a member of the phosphatase binding protein family. AB - P84 is a neuronal membrane glycoprotein that promotes the attachment and neurite outgrowth of cultured murine cerebellar cells. The heterophilic adhesive properties of P84 and its localization at sites of synaptogenesis suggest that it may be involved in regulation of synapse formation or maintenance. P84 is expressed in subsets of neurons throughout the CNS. By cloning the cDNA encoding murine P84, we have discovered that this molecule is a member of a family of phosphatase-binding proteins and is identical to the murine SHPS-1 cDNA. Here we report the cloning of two alternatively spliced forms of P84 and describe its localization within the CNS by in situ hybridization. PMID- 9348340 TI - DNA replication and postreplication mismatch repair in cell-free extracts from cultured human neuroblastoma and fibroblast cells. AB - DNA synthesis and postreplication mismatch repair were measured in vitro using cell-free extracts from cultured human SY5Y neuroblastoma and WI38 fibroblast cells in different growth states. All extracts, including differentiated SY5Y and quiescent WI38 fibroblasts, catalyzed SV40 origin-dependent DNA synthesis, totally dependent on SV40 T-antigen. Thus, although differentiated neuroblastoma and quiescent fibroblasts cells were essentially nondividing, their extracts were competent for DNA replication using DNA polymerases delta, alpha, and possibly epsilon, with proliferating cell nuclear antigen. Nonreplicative DNA synthesis and lesion bypass by either alpha- or beta-polymerases were detected independently in extracts using primed or gapped single-stranded DNA templates. Long-patch postreplication mismatch repair was measured for the first time in neuroblastoma cell-free extracts. Extracts from subconfluent and high-density SY5Y cells catalyzed postreplication mismatch repair with efficiencies comparable to those of HeLa cell extracts. No significant differences were observed in repair between SY5Y differentiated and undifferentiated cell extracts. Mismatch repair efficiencies were threefold lower in extracts from subconfluent WI38 cells, and repair in WI38 quiescent cells was fourfold less than in subconfluent cells, suggesting that mismatch repair may be regulated. The spectrum of mismatch repair in SY5Y extracts closely resembled the mismatch removal specificities of HeLa extracts: T . G and G . G mismatches were repaired most efficiently; C . A, A . A, A . G and a five-base loop were repaired with intermediate efficiency; repair of G . A, C . C, and T . T mismatches was extremely inefficient. PMID- 9348341 TI - Nociceptin inhibits T-type Ca2+ channel current in rat sensory neurons by a G protein-independent mechanism. AB - Nociceptin (orphanin FQ) is a novel, opioid-like, heptadecapeptide that is an endogenous ligand for the opioid receptor-like (ORL1) receptor. Unlike classical opioids, nociceptin can produce hyperalgesia when injected intracerebroventricularly into mice. Despite this, nociceptin has been reported to decrease transmitter release, activate an inwardly rectifying K+ conductance, and suppress high-voltage-activated Ca2+ channel conductances (HVA gCa) in much the same way as micro-, delta-, and kappa-opioids. We report an action of nociceptin that is not shared by morphine: the suppression of low-voltage activated, transient calcium (barium) current (IBa,T) in acutely dissociated rat dorsal root ganglion (DRG) neurons (EC50 = 100 nM). This effect was reflected as inhibition of bursts of action potentials that can be evoked in "medium-sized" DRG neurons. Experiments with GTP-gamma-S (100 microM), GDP-beta-S (2 mM), or aluminum fluoride (AlF3) (100 microM) in the patch pipette failed to provide evidence for G-protein involvement in nociceptin-induced IBa,T suppression. By contrast, both morphine and nociceptin suppressed HVA gCa, and the latter response was affected by intracellular GTP-gamma-S, GDP-beta-S, and AlF3 in ways that confirmed G-protein involvement. The selective effect of nociceptin on IBa,T may therefore be relevant to understanding why its behavioral actions differ from those of other opioids. This G-protein-independent effect of the action of nociceptin may reflect a new general mechanism of action for opioid peptides within the nervous system. PMID- 9348342 TI - Activity-dependent calcium sequestration in dendrites of hippocampal neurons in brain slices. AB - Synaptic activity-dependent changes in the spatio-temporal distribution of calcium ions regulate important neuronal functions such as dendritic integration and synaptic plasticity, but the processes that terminate the free Ca2+ transients associated with these changes remain unclear. We have characterized at the electron microscopic level the intracellular compartments involved in buffering free Ca2+ transients in dendritic cytoplasm of CA3 neurons by measuring the larger changes in the concentrations of total Ca that persist for several minutes after neuronal activity. Quantitative energy-dispersive x-ray microanalysis of cryosections from hippocampal slice cultures rapidly frozen 3 min after afferent synaptic activity identified a subset of dendritic endoplasmic reticulum (ER) as a high-capacity Ca2+ buffer. Calcium sequestration by cisterns of this subset of ER was graded, reversible, and dependent on a thapsigargin sensitive Ca2+-ATPase. Sequestration was so robust that after repetitive high frequency stimulation the Ca content of responsive ER cisterns increased as much as 20-fold. These results demonstrate that a subpopulation of ER is the major dendritic Ca sequestration compartment in the minutes after neuronal activity. PMID- 9348343 TI - Mechanoelectrical transduction and adaptation in hair cells of the mouse utricle, a low-frequency vestibular organ. AB - Hair cells of inner ear organs sensitive to frequencies above 10 Hz adapt to maintained hair bundle deflections at rates that reduce their responses to lower frequencies. Mammalian vestibular organs detect head movements at frequencies well below 10 Hz. We asked whether hair cells of the mouse utricle adapt, and if so, whether the adaptation was similar to that in higher frequency organs such as the frog saccule. Whole-cell transduction currents were recorded from hair cells in the epithelium of the mouse utricle. Hair bundles were deflected by a fluid jet or a stiff probe. The transduction currents evoked by step deflections adapted over 10-100 msec. The mean operating range was 1.5 micron (deflection of the tip of the bundle), approximately threefold larger than in frog saccule. Taller and more compact bundles of the mouse utricle account for this difference. As in frog saccular hair cells, adaptation shifted the current-deflection (I(X)) relation along the deflection axis. These adaptive shifts had time constants of 10-20 msec and reached 60-80% of stimulus amplitude. The adaptive shift and voltage-dependent bundle movement are consistent with the motor model of adaptation. When the fluid jet was used, adaptation also broadened the I(X) relation and reduced the maximum current. Adaptation attenuated the transduction currents evoked by sinusoidal bundle deflections below 5 Hz, within the frequency range of the utricle, but because it was incomplete, substantial responses remained. Moreover, the adaptive shift mechanism preserves sensitivity even in the presence of large stimuli that would otherwise saturate transduction. PMID- 9348344 TI - Critical role of TrkB and brain-derived neurotrophic factor in the differentiation and survival of retinal pigment epithelium. AB - In the vertebrate eye, the retinal pigment epithelium (RPE) and the neural retina arise from a single layer of neuroectoderm. Factors influencing the differentiation of retinal neurons have been identified; however, little is known about molecules directing the differentiation of the RPE. Here we have found that the neurotrophin brain-derived neurotrophic factor (BDNF) plays an autocrine role in the differentiation and survival of Xenopus laevis RPE. Fluorescent in situ hybridization studies showed a precise co-expression of BDNF and its receptor trkB in the retinal neuroepithelium and actively differentiating RPE; in vitro studies demonstrated survival- and differentiation-promoting effects in serum free explants and dissociated cultures. When a dominant negative mutant of the trkB receptor was expressed in developing embryos, severe arrest of RPE differentiation was seen with persistence of nestin- and Notch-positive neuroblasts. PMID- 9348345 TI - Mutant superoxide dismutase-1-linked familial amyotrophic lateral sclerosis: molecular mechanisms of neuronal death and protection. AB - Mutations in human Cu/Zn superoxide dismutase-1 (SOD) cause approximately 20% of cases of familial amyotrophic lateral sclerosis (FALS). We investigated the mechanism of mutant SOD-induced neuronal degeneration by expressing wild-type and mutant SODs in neuronal cells by means of infection with replication-deficient recombinant adenoviruses. Expression of two FALS-related mutant SODs (A4V and V148G) caused death of differentiated PC12 cells, superior cervical ganglion neurons, and hippocampal pyramidal neurons. Cell death included many features typical of apoptosis. Death could be prevented by copper (Cu2+) chelators, Bcl-2, glutathione, vitamin E, and inhibitors of caspases. Mutant SOD-expressing PC12 cells had higher rates of superoxide (O2-) production under a variety of conditions. The results support the hypothesis that mutant SOD induced neurodegeneration is associated with disturbances of neuronal free radical homeostasis. PMID- 9348346 TI - Cerebellar disorganization characteristic of reeler in scrambler mutant mice despite presence of reelin. AB - Analysis of the molecular basis of neuronal migration in the mammalian CNS relies critically on the discovery and identification of genetic mutations that affect this process. Here, we report the detailed cerebellar phenotype caused by a new autosomal recessive neurological mouse mutation, scrambler (gene symbol scm). The scrambler mutation results in ataxic mice that exhibit several neuroanatomic defects reminiscent of reeler. The most obvious of these lies in the cerebellum, which is small and lacks foliation. Granule cells, although normally placed in an internal granule cell layer, are greatly reduced in number ( approximately 20% of normal). Purkinje cells are also reduced in number, and the majority are located ectopically in deep cerebellar masses. There is a small population of Purkinje cells ( approximately 5% of the total) that occupy a Purkinje cell layer between the molecular and granule cell layers. Despite this apparent disorganization of Purkinje cells, zebrin-positive and zebrin-negative parasagittal zones can be delineated. The ectopic masses of Purkinje cells are bordered by the extracellular matrix protein tenascin and by processes containing glial fibrillary acidic protein. Antibodies specific for these proteins also identify a novel midline raphe structure in both scrambler and reeler cerebellum that is not present in wild-type mice. Thus, in many respects, the scrambler cerebellum is identical to that of reeler. However, the scrambler locus has been mapped to a site distinct from that of reelin (Reln), the gene responsible for the reeler defect. Here we find that there are normal levels of Reln mRNA in scrambler brain and that reelin protein is secreted normally by scrambler cerebellar cells. These findings imply that the scrambler gene product may function in a molecular pathway critical for neuronal migration that is tightly linked to, but downstream of, reelin. PMID- 9348347 TI - Targeted overexpression of the neurite growth-associated protein B-50/GAP-43 in cerebellar Purkinje cells induces sprouting after axotomy but not axon regeneration into growth-permissive transplants. AB - B-50/GAP-43 is a nervous tissue-specific protein, the expression of which is associated with axon growth and regeneration. Its overexpression in transgenic mice produces spontaneous axonal sprouting and enhances induced remodeling in several neuron populations (; ). We examined the capacity of this protein to increase the regenerative potential of injured adult central axons, by inducing targeted B-50/GAP-43 overexpression in Purkinje cells, which normally show poor regenerative capabilities. Thus, transgenic mice were produced in which B-50/GAP 43 overexpression was driven by the Purkinje cell-specific L7 promoter. Uninjured transgenic Purkinje cells displayed normal morphology, indicating that transgene expression does not modify the normal phenotype of these neurons. By contrast, after axotomy numerous transgenic Purkinje cells exhibited profuse sprouting along the axon and at its severed end. Nevertheless, despite these growth phenomena, which never occurred in wild-type mice, the severed transgenic axons were not able to regenerate, either spontaneously or into embryonic neural or Schwann cell grafts placed into the lesion site. Finally, although only a moderate Purkinje cell loss occurred in wild-type cerebella after axotomy, a considerable number of injured transgenic neurons degenerated, but they could be partially rescued by the different transplants placed into the lesion site. Thus, B-50/GAP-43 overexpression substantially modifies Purkinje cell response to axotomy, by inducing growth processes and decreasing their resistance to injury. However, the presence of this protein is not sufficient to enable these neurons to accomplish a full program of axon regeneration. PMID- 9348348 TI - Preferential termination of corticorubral axons on spine-like dendritic protrusions in developing cat. AB - The formation of synaptic contacts is a crucial event during neural development and is thought to be achieved by complex interactions between incoming axons and the neurons in the target. We have focused on spine-like dendritic protrusions (SLDPs), which are transient pleomorphic protrusive structures seen in developing brains. Although the functional significance of SLDPs remains unknown, accumulating in vitro evidence suggests that the SLDP plays an important role in synaptogenetic interactions with axons. As a test of this idea, the present study was performed to examine whether the SLDPs are the preferential sites of synapse formation in vivo. The ultrastructure of biocytin-labeled corticorubral (CR) terminals was examined in serial thin sections during the period of synaptogenesis in newborn cats. We found that a major proportion (86%) of the CR synapses was formed on SLDPs. The presynaptic terminals were often invaginated by fine processes extending from the tips of SLDPs. Synaptic structures presumably of cortical origin were also found on SLDPs of HRP-labeled rubrospinal cells, suggesting that SLDPs postsynaptic to labeled CR terminals originate at least in part from rubrospinal cells. Taken together, these results indicate that SLDPs may represent preferred sites of synapse formation and support the notion that SLDPs play a role in synaptogenic interactions during brain development. PMID- 9348349 TI - Excitotoxicity in the enteric nervous system. AB - Glutamate, the major excitatory neurotransmitter in the CNS, is also an excitatory neurotransmitter in the enteric nervous system (ENS). We tested the hypothesis that excessive exposure to glutamate, or related agonists, produces neurotoxicity in enteric neurons. Prolonged stimulation of enteric ganglia by glutamate caused necrosis and apoptosis in enteric neurons. Acute and delayed cell deaths were observed. Glutamate neurotoxicity was mimicked by NMDA and blocked by the NMDA antagonist D-2-amino-5-phosphonopentanoate. Excitotoxicity was more pronounced in cultured enteric ganglia than in intact preparations of bowel, presumably because of a reduction in glutamate uptake. Glutamate immunoreactive neurons were found in cultured myenteric ganglia, and a subset of enteric neurons expressed NMDA (NR1, NR2A/B), AMPA (GluR1, GluR2/3), and kainate (GluR5/6/7) receptor subunits. Glutamate receptors were clustered on enteric neurites. Stimulation of cultured enteric neurons by kainic acid led to the swelling of somas and the growth of varicosities ("blebs") on neurites. Blebs formed close to neurite intersections and were enriched in mitochondria, as revealed by rhodamine 123 staining. Kainic acid also produced a loss of mitochondrial membrane potential in cultured enteric neurons at sites where blebs tended to form. These observations demonstrate, for the first time, excitotoxicity in the ENS and suggest that overactivation of enteric glutamate receptors may contribute to the intestinal damage produced by anoxia, ischemia, and excitotoxins present in food. PMID- 9348350 TI - Defective learning in mutants of the Drosophila gene for a regulatory subunit of cAMP-dependent protein kinase. AB - Disruptions of a Drosophila gene encoding a regulatory subunit of cAMP-dependent protein kinase homologous to mammalian RIbeta (dPKA-RI) were targeted to the first (noncoding) exon of dPKA-RI via site-selected P element mutagenesis. Flies homozygous for either of two mutant alleles showed specific defects in olfactory learning but not in subsequent memory decay. In contrast, olfactory acuity and shock reactivity, component behaviors required for normal odor avoidance learning, were normal in these mutants. Northern and Western blot analyses of mRNA and protein extracted from adult heads have revealed a complex lesion of the PKA-RI locus, including expression of a novel product and over- or underexpression of wild-type products in mutants. Western blot analysis revealed reductions in RI protein in mutants. PKA activity in the absence of exogenous cAMP also was significantly higher than normal in homogenates from mutant adult heads. These two mutant alleles failed to complement each other for each of these phenotypic defects, eliminating second-site mutations as a possible explanation. These results establish a role for an RI regulatory subunit of PKA in Pavlovian olfactory conditioning. PMID- 9348351 TI - The olivocerebellar projection mediates ibogaine-induced degeneration of Purkinje cells: a model of indirect, trans-synaptic excitotoxicity. AB - Ibogaine, an indole alkaloid that causes hallucinations, tremor, and ataxia, produces cerebellar neurotoxicity in rats, manifested by degeneration of Purkinje cells aligned in narrow parasagittal bands that are coextensive with activated glial cells. Harmaline, a closely related alkaloid that excites inferior olivary neurons, causes the same pattern of Purkinje cell degeneration, providing a clue to the mechanism of toxicity. We have proposed that ibogaine, like harmaline, excites neurons in the inferior olive, leading to sustained release of glutamate at climbing fiber synapses on Purkinje cells. The objective of this study was to test the hypothesis that increased climbing fiber activity induced by ibogaine mediates excitotoxic Purkinje cell degeneration. The inferior olive was pharmacologically ablated in rats by a neurotoxic drug regimen using 3 acetylpyridine, and cerebellar damage attributed to subsequent administration of ibogaine was analyzed using immunocytochemical markers for neurons and glial cells. The results show that ibogaine administered after inferior olive ablation produced little or no Purkinje cell degeneration or glial activation. That a lesion of the inferior olive almost completely prevents the neurotoxicity demonstrates that ibogaine is not directly toxic to Purkinje cells, but that the toxicity is indirect and dependent on integrity of the olivocerebellar projection. We postulate that ibogaine-induced activation of inferior olivary neurons leads to release of glutamate simultaneously at hundreds of climbing fiber terminals distributed widely over the surface of each Purkinje cell. The unique circuitry of the olivocerebellar projection provides this system with maximum synaptic security, a feature that confers on Purkinje cells a high degree of vulnerability to excitotoxic injury. PMID- 9348352 TI - Social stress in hamsters: defeat activates specific neurocircuits within the brain. AB - During an agonistic encounter, subordinate male hamsters display defensive and submissive postures and show increased secretion of glucocorticoids, whereas dominant males do not. To determine whether specific neuronal pathways are activated during the behavioral and neuroendocrine responses of subordinate males, expression of c-fos mRNA within the brains of subordinate males was compared with the pattern in dominant males after fighting. After 1 week of handling, pairs of hamsters were either swapped between cages (handled control males), or were allowed to interact for 30 min [dominant (DOM) males and subordinate (SUB) males]. A second group of control animals that received no handling or social stimulation (unhandled control males) were also included. After testing, all animals were killed by decapitation, their brains were removed for c-fos in situ hybridization, and trunk blood was collected for analysis of plasma cortisol and corticosterone levels. Exposure of males to their partner's cage for 30 min resulted in increased expression of c-fos mRNA in multiple brain regions. In addition, fighting increased c-fos expression in the medial amygdaloid nucleus of both DOM and SUB males as well as having more selective effects. In DOM males, c-fos expression was elevated within the supraoptic nucleus of the hypothalamus. In SUB males, c-fos expression increased within a multitude of brain areas, including cingulate cortex, lateral septum, bed nucleus of the stria terminalis, medial preoptic area, several hypothalamic nuclei, central amygdaloid nucleus, amygdalohippocampal area, dorsal periaqueductal gray, dorsal raphe, cuneiform nucleus, and locus coeruleus. These findings are discussed in relation to neurocircuits associated with behavioral arousal and stress. PMID- 9348353 TI - An initial, three-day-long treatment with alcohol induces a long-lasting phenomenon of selective tolerance in the activity of the rat hypothalamic pituitary-adrenal axis. AB - We determined whether an initial alcohol challenge induced long-lasting changes in the activity of the hypothalamic-pituitary-adrenal (HPA) axis. Adult male rats received intragastric injections of the vehicle or a moderately intoxicating dose of alcohol (3.0 gm/kg) daily for 3 d. When animals were acutely challenged with alcohol 3-12 d later, their ACTH and corticosterone responses were significantly blunted, compared with that of vehicle-pretreated rats. In contrast, exposure to mild electric foot shocks induced a pattern of ACTH secretion that was comparable in animals administered alcohol or the vehicle previously, indicating a lack of cross-tolerance. No significant differences were observed in pituitary responsiveness to corticotropin-releasing factor or vasopressin in rats pretreated with the vehicle or alcohol. The influence of the initial drug treatment was not mimicked by exposure to foot shocks, nor was it prevented by administering a potent corticotropin-releasing factor antagonist to block the elevations in plasma ACTH and corticosterone induced by this initial treatment. Finally, we found that rats injected initially with the vehicle and challenged subsequently with alcohol exhibited the expected increased neuronal activation (measured by the upregulation of steady-state mRNA and protein levels of immediate early genes) in the paraventricular nucleus of their hypothalamus. In contrast, this response was markedly decreased in animals exposed previously to the drug. To our knowledge, this is the first report that exposure to a stress (i.e., alcohol), although not immediately altering the response of the HPA axis to this particular signal, induces a selective tolerance that is both slow to develop and long-lasting. The primary mechanism mediating the ability of an initial drug treatment to decrease subsequent responses of the HPA axis to a second drug challenge seems to be the inability of hypothalamic neurons to respond adequately to this second challenge. PMID- 9348354 TI - Postexcitatory inhibition of the crayfish lateral giant neuron: a mechanism for sensory temporal filtering. AB - Crayfish escape from threats by either giant neuron-mediated "reflex" tail flexions that occur with very little delay but do not allow for much sensory guidance of trajectory or by "nongiant" tail flexion responses that allow for sensory guidance but occur much less promptly. Thus, when a stimulus occurs, the nervous system must make a rapid assessment of whether to use the faster reflex system or the slower nongiant one. It does this on the basis of the abruptness of stimulus onset; only stimuli of very abrupt onset trigger giant-mediated responses. We report here that stimuli which excite the lateral giant (LG) command neurons for one form of reflex escape also produce a slightly delayed postexcitatory inhibition (PEI) of the command neurons. As a result, only stimuli that become strong enough to excite the command neurons to firing threshold before the onset of PEI, within a few milliseconds of stimulus onset, can cause giant-mediated responses. This inhibition is directed to distal dendrites of the LG neurons, which allows for some location specificity of PEI within the sensory field of a single hemisegment. PMID- 9348355 TI - Functional properties of perigeniculate inhibition of dorsal lateral geniculate nucleus thalamocortical neurons in vitro. AB - The properties of the inhibitory influence of neurons in the perigeniculate (PGN) nucleus on thalamocortical cells were examined with intracellular recordings in the ferret geniculate slice maintained in vitro. Activation of PGN neurons with the local application of glutamate caused IPSPs in thalamocortical neurons that were mediated by both GABAA and GABAB receptors, as well as the activation of spindle waves. With low intensity stimulation of the PGN, local application of bicuculline to the dorsal lateral geniculate nucleus (LGNd) strongly inhibited evoked and spindle-associated IPSPs, indicating that these are largely mediated by GABAA receptors. The generation of GABAB receptor-mediated IPSPs in thalamocortical cells that were large enough to generate rebound low threshold Ca2+ spikes required substantially increased activation of the PGN with glutamate. The activation of synchronous bicuculline-induced slowed oscillations in thalamocortical neurons required the block of GABAA receptors in the LGNd as well as in the PGN. These results indicate that bursts of action potentials in PGN neurons can result in the activation of both GABAA and GABAB receptors in thalamocortical neurons, with the strong activation of GABAB receptors requiring an intense, simultaneous discharge of a number of PGN neurons. Functionally, these results suggest that PGN neurons inhibit thalamocortical cells preferentially through the activation of GABAA receptors, although the strong activation of GABAB receptors may occur under pathological conditions and contribute to the generation of abnormal, synchronous slow oscillations. PMID- 9348357 TI - Photoperiodically driven changes in Fos expression within the basal tuberal hypothalamus and median eminence of Japanese quail. AB - The rapid photoperiodic response in Japanese quail is so precise that it allows neural analyses of how photoperiodic information is transduced into an endocrine response. After transfer from short [SD; 6L:18D (6:18 hr light/dark cycle)] to long (LD; 20L:4D) days, luteinizing hormone (LH) first rises 20 hr after dawn. Using Fos immunocytochemistry, we examined the basal tuberal hypothalamus (BtH) to determine the relationship between brain cell activation and the first endocrine changes. Two separate cell populations within the BtH expressed Fos like immunoreactivity (FLI) by hour 18 of the first LD. Importantly, this activation occurred before the LH rise. Median eminence activation appeared within glial cells, whereas activated infundibular nucleus cells were neuronal, providing support to the view that gonadotropin-releasing hormone (GnRH) release can be controlled at the terminals by glia. The FLI induction parallels LH changes, suggesting that gene expression may be involved in events preceding photostimulation and is the earliest photoperiodically stimulated physiological change yet reported. Additional experiments provided further support for this hypothesis. First, photoperiodically induced activation is not a result peculiar to castrates because intact birds displayed similar results. Second, the critical length of 14 hr of light had to be exceeded to cause both BtH activation and a LH rise 30 hr from dawn. Finally, valuable evidence of the response specificity was provided by using a unique property of the quail photoperiodic clock in which exposure to 10L:26D, but not 10L:14D, causes photoinduction. The 36 hr paradigm increased both plasma LH and BtH activation. PMID- 9348356 TI - Inhibitory interactions between perigeniculate GABAergic neurons. AB - Perigeniculate neurons form an interactive sheet of cells that inhibit one another as well as thalamocortical neurons in the dorsal lateral geniculate nucleus (LGNd). The inhibitory influence of the GABAergic neurons of the perigeniculate nucleus (PGN) onto other PGN neurons was examined with intracellular recordings in vitro. Intracellular recordings from PGN neurons during the generation of spindle waves revealed barrages of EPSPs and IPSPs. The excitation of local regions of the PGN with the local application of glutamate resulted in activation of IPSPs in neighboring PGN neurons. These IPSPs displayed an average reversal potential of -77 mV and were blocked by application of bicuculline methiodide or picrotoxin, indicating that they are mediated by GABAA receptors. In the presence of GABAA receptor blockade, the activation of PGN neurons with glutamate could result in slow IPSPs that were mediated by GABAB receptors in a subset (40%) of cells. Similarly, application of specific agonists muscimol and baclofen demonstrated that PGN neurons possess both functional GABAA and GABAB receptors. Examination of the axon arbors of biocytin-filled PGN neurons often revealed the presence of beaded axon collaterals within the PGN, suggesting that this may be an anatomical substrate for PGN to PGN inhibition. Functionally, activation of inhibition between PGN neurons could result in a shortening or a complete abolition of the low threshold Ca2+ spike or an inhibition of tonic discharge. We suggest that the mutual inhibition between PGN neurons forms a mechanism by which the excitability of these cells is tightly controlled. The activation of a point within the PGN may result in the inhibition of neighboring PGN neurons. This may be reflected in the LGNd as a center of inhibition surrounded by an annulus of disinhibition, thus forming a "center surround" mechanism for thalamic function. PMID- 9348358 TI - Unilateral lesions of the dorsal striatum in rats disrupt responding in egocentric space. AB - Rats were trained in a specially designed, multichoice operant chamber on a visual choice reaction time task designed to assess performance on each side of the rat's body. The task required animals to sustain a nose poke in a central hole, until a brief light stimulus was presented in either of two holes that were located on the same side of the box. Once the rats were trained to perform the task to both sides independently they received unilateral injections of quinolinic acid into the dorsal striatum. Postoperatively, lesioned animals were impaired when performing the task on the side contralateral to the lesion. The time taken to initiate contralateral responses was increased. Contralateral responses were also exclusively biased toward the nearer of the two response locations, regardless of the location of the stimulus. This was interpreted as a specific impairment in generating responses in contralateral space. In contrast, no comparable deficit was seen when the animals performed the task on the side ipsilateral to the lesion. Additional postoperative challenges, in which response options were presented bilaterally, showed this response deficit to be defined in egocentric coordinates, with the severest response deficits for the most contralateral locations. PMID- 9348360 TI - Your journal-analysis and trends PMID- 9348359 TI - A journal of evidence PMID- 9348361 TI - Nonsurgical management of chronic pelvic pain. PMID- 9348362 TI - Clinical trial of the uterine thermal balloon for treatment of menorrhagia. AB - STUDY OBJECTIVES: To evaluate the safety and efficacy of thermal balloon endometrial ablation in women with menorrhagia, and to identify factors influencing outcome. DESIGN: Prospective, observational study (Canadian Task Force classification II-2). SETTING: Three Canadian university-affiliated teaching hospitals. PATIENTS: One hundred twenty-one women suffering from menorrhagia serious enough to make them candidates for endometrial ablation or hysterectomy. Patients without obvious structural or (pre)malignant abnormalities were included if their uterine cavities sounded to less than 12 cm, they were in good health, and had undergone hysteroscopy or pelvic ultrasound and endometrial biopsy within 6 months and had a normal Papanicolaou smear within 1 year. INTERVENTIONS: A balloon catheter was placed through the cervix and after inflation in the endometrial cavity with 5% dextrose in water, was heated to 87 +/- 5 degrees C. Two-thirds of patients avoided general anesthesia and very few required cervical dilatation to admit the 4.5-mm diameter catheter. Balloon pressures were 90 to 140 mm Hg in 13 patients; pressures between 140 and 190 mm Hg were well tolerated by the rest. Nineteen women underwent 12 minutes of therapy, and the rest had 8-minute sessions. MEASUREMENTS AND MAIN RESULTS: No intraoperative complications occurred, and minor postoperative morbidity occurred in 4% of patients. Preoperative and postoperative bleeding was assessed by pad counts and patient self-reports. The degree of dysmenorrhea was recorded similarly. A paired t test was used to compare pretreatment with posttreatment pad counts. A Wilcoxon signed rank test was employed to evaluate the effect of treatment on dysmenorrhea. The effects on outcome of several independent variables were analyzed by multiple and logistic regression. Success of the procedure was constant over the year (range 86-90%). Treatment led to significant decreases in menstrual flow, duration, and pain (p <0.0001). No significant effects of parity, uterine position or cavity depth, timing, or various endometrial-thinning regimens were found. Increasing age was significantly associated with increased odds of success (p < 0.05). Excluding the 19 women who underwent 12 minutes of therapy did not change statistical results, whereas excluding the 13 treated with balloon pressures less than 140 mm Hg improved the results. Conclusion. The facts that bleeding and dysmenorrhea were significantly reduced by thermal balloon endometrial ablation, that no intraoperative complication occurred, and that postoperative morbidity was minimal, lead us to conclude that this is potentially a safe and effective technique. Larger studies and longer follow-up are required to substantiate this impression. PMID- 9348364 TI - Polypectomy and endometrial resection in postmenopausal patients. AB - STUDY OBJECTIVE: To evaluate the therapeutic efficacy of polypectomy associated with endometrial resection for the treatment of polyps in postmenopausal women. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Private urban hospital with facilities for endoscopic surgery. PATIENTS: Sixty six women with endometrial polyps. INTERVENTIONS: Sixty-four of the 66 patients underwent polypectomy followed by endometrial resection. One patient had hysterectomy because endometrial biopsy showed serous papillary carcinoma. MEASUREMENTS AND MAIN RESULTS: No major complications were associated with the procedure. Histopathology showed hyperplasia in most polyps. One patient had a papillary uterine carcinoma in the polyp that was not detected by preoperative endometrial biopsy. The women were followed with transvaginal sonography for 1 year after surgery. CONCLUSION: Polypectomy followed by endometrial resection is a very low-risk procedure for postmenopausal patients. There was no recurrence after the first year in women who completed follow-up. PMID- 9348363 TI - Preventing hyponatremic encephalopathy: comparison of serum sodium and osmolality during operative hysteroscopy with 5.0% mannitol and 1.5% glycine distention media. AB - STUDY OBJECTIVE: To determine whether isotonic 5.0% mannitol is superior to 1.5% glycine in preventing development of hyponatremic encephalopathy. DESIGN: Prospective, comparative study (Canadian Task Force classification II=2). SETTING: Gynecology department of a community hospital. PATIENTS: One hundred twenty-two women undergoing operative hysteroscopy. INTERVENTIONS: Eighteen blood serum chemical indicators analyzed preoperatively and postoperatively in 61 women undergoing operative hysteroscopy with 1. 5% glycine (group 1) were compared with those of 61 women having similar surgery with 5.0% mannitol (group 2). Fluid deficit (difference between input and output volume of distention fluid) was recorded, and differences between presurgical and postsurgical indicators of the two groups (mean difference score) were compared. MEASUREMENTS AND MAIN RESULTS: Mean +/- SEM sodium difference scores of groups 1 and 2 were -1.73 +/- 0.42 mEq/L (range -7.00 to 2.00 mEq/L) and -5.04 +/- 1.07 mEq/L (range -36.00 to 3.00 mEq/L), respectively (p <0.01). Serum osmolality difference scores were -6. 88 +/ 1.36 mmol/L (range -13.00 to -1.00 mmol/L) and -1.87 +/- 0.35 mmol/L (range -3 to 15 mmol/L), respectively (p <0.01). Distention fluid deficits were 0.435 +/- 0.071 L (range 0-2.448 L) and 0.473 +/- 0.084 L (range 0-3.640 L), respectively (p = 0.862). Two women (3.4%) in group 1 and five (8.2%) in group 2 developed postoperative asymptomatic dilutional hyponatremia (p = 0.211), which was the only complication. Two of the five women in group 2 developed severe dilutional hyponatremia. CONCLUSION: We found that 5.0% mannitol distention fluid produces greater postoperative dilutional hyponatremia than 1.5% glycine, but hypo osmolality does not occur with mannitol. Its use should lessen the risk of hyponatremic encephalopathy. PMID- 9348365 TI - Association of positive Chlamydia trachomatis and Chlamydia pneumoniae immunoglobulin-gamma titers with increasing age. AB - STUDY OBJECTIVES: To use Chlamydia trachomatis immunoglobulin-gamma (IgG) titers to investigate the possibility of their association with ovarian cancer, and to evaluate the effectiveness of this titer in algorithmic protocols in infertility. DESIGN: Prospective, age-matched, pilot study (Canadian Task Force classification II-2). SETTING: University and university-affiliated office practice. PATIENTS: The original 30 patients were seen for follow-up of ovarian cancer (19) and at yearly examination for nonmalignant disease (10). An additional group of 21 women seen for pelvic pain and infertility was added to clarify questions that arose during the study. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Chlamydia trachomatis IgG titers were positive in 15 (79%) of 19 of women with ovarian cancer, 9 (90%) of 10 age-matched controls, and 14 (67%) of 21 patients with infertility and pain. When analyzed by age, 4 (40%) of 10 patients under 30 years and 34 (85%) of 40 patients 30 years of age or older had positive titers (p = 0.007). Of 21 women with positive Chlamydia pneumoniae titers, 17 (81 %) had positive C. trachomatis titers, and 17 (85%) of 20 with positive C. trachomatis titers had positive C. pneumoniae titers. CONCLUSION: The test kit used in this study may not be adequate in older patients due to cross-reaction with C. pneumoniae titers. Further evaluation of C. trachomatis IgG titers as a marker in the study of ovarian cancer will require titers that are more specific than those we used. Although these titers may be useful as an immunologic screening marker in infertile patients, results should be interpreted with caution. A positive test may not be evidence of C. trachomatis infection and is not an indication for specific therapy. Successful use of some currently available C. trachomatis IgG titers in algorithms for infertility may be related to a patient's age. PMID- 9348366 TI - Conscious pain mapping. AB - STUDY OBJECTIVE: To present the technique and usefulness of conscious pain mapping. DESIGN: Prospective, observational study (Canadian Task Force classification II-2). SETTING: Gynecology departments of a university-affiliated hospital and a private community hospital. PATIENTS: Fifty consecutive women undergoing diagnostic microlaparoscopy. INTERVENTIONS: Conscious pain mapping was performed in all 50 women. MEASUREMENTS AND MAIN RESULTS: Conscious pain mapping helped to identify foci of chronic pelvic pain. The appendix and pelvic adhesions accounted for a significant amount of pelvic pain in these women. CONCLUSIONS: Conscious pain mapping helps to uncover sites of pelvic pain that might not be identified during traditional laparoscopy under general anesthesia. PMID- 9348367 TI - A protocol for conscious sedation in microlaparoscopy. AB - To establish a protocol for conscious sedation in microlaparoscopy, we conducted a prospective, observational study of 74 women undergoing the procedure under local anesthesia with conscious sedation for the evaluation and treatment of chronic pelvic pain. Our protocol for conscious sedation allowed us to perform diagnostic microlaparoscopy under local anesthesia in all 74 women and operative microlaparoscopy in 52 (70.2%). This procedure is a safe and effective alternative to general anesthesia during microlaparoscopy in selected patients. PMID- 9348368 TI - Direct laparoscopic cannula insertion at the left upper quadrant. AB - We evaluated the efficacy and safety of direct left upper quadrant (LUQ) cannula insertion for laparoscopic surgery in 23 women with prior pelvic surgery, compared with direct umbilical cannula insertion in a control group of 81 patients. Generally, the laparoscope was retained at the LUQ site throughout the operative procedure. Cannula insertions at the LUQ were successful in the first attempt in 22 patients, compared with a single successful attempt in 78 of 81 umbilical insertions. Nine women had anterior abdominal wall adhesions that extended to the umbilical area. Seven had either a prior midline (1) or Pfannenstiel (6) incision; all seven had direct LUQ cannula insertions. Two patients with umbilical adhesions had no prior surgery. Of the three complications, two were related to cannula insertions and both were in the control group. There were no bowel injuries. More experience is required to prove that LUQ cannula insertion accomplishes its intended aim of avoiding bowel or omental injuries due to adhesions in women with prior abdominopelvic surgery, but initial results were favorable. PMID- 9348369 TI - Laparoscopic lateral ovarian transposition before radiation treatment of Hodgkin disease. AB - More than one-half of women treated with pelvic radiation therapy for malignant disease experience premature ovarian failure. Preservation of ovarian function by repositioning the ovaries out of the irradiation field is suggested in all women of reproductive age. This repositioning generally is done by moving the ovaries either medially so they are posterior to the uterus, or laterally so they are in the paracolic gutters. Laparoscopic medial transposition has been reported, with mixed results. A woman underwent successful laparoscopic lateral transposition before irradiation for stage IIIa Hodgkin disease. A review of published cases suggests that this is preferable to medial transposition. PMID- 9348370 TI - Laparoscopic management of pelvic pathology during pregnancy. AB - Advanced operative laparoscopy is being performed increasingly for various indications and in diverse patient populations, including gravid women. In the United States approximately 1.6% to 2.2% of pregnant women require nonobstetric surgery for abdominal and pelvic pathology. Increasing numbers of case reports suggest the feasibility and safety of operative laparoscopy during pregnancy. We identified certain management issues specific to these procedures based on our experience with nine cases of operative laparoscopy in women with gestations up to 22 weeks. PMID- 9348371 TI - Successful laparoscopic removal of huge ovarian cysts. AB - Laparoscopic management of huge adnexal masses is controversial mainly because of the fear of finding a malignancy and because of technical difficulties. We successfully removed two huge ovarian cysts using only an operative laparoscope, a 10-mm infraumbilical, and three 5-mm suprapubic cannulas. We believe that minimally invasive surgery can be performed safely and effectively in carefully selected patients with extremely large, abdominopelvic masses. PMID- 9348372 TI - What is evidence-based medicine? AB - This is the first in a series of articles on evidence-based medicine. This series is intended to act as a primer for the novice who needs to understand more about the applications of this exciting new concept to the field of gynecology. The first article provides an overview of the field; future installments will discuss specific applications and techniques with regard to evidence-based medicine, and the assets and liabilities of putting this method into practice. It is hoped that the reader will find the material both practical and provocative. PMID- 9348373 TI - Minimal access and open surgery. Competition or integration? PMID- 9348374 TI - Multicentric experience of the Belgian Group for Endoscopic Surgery (BGES) with endoscopic adrenalectomy. AB - BACKGROUND: Adrenalectomy is not a frequent operation. Therefore the newly developed laparoscopic approach is sporadically performed by surgeons dealing with endocrine disorders. METHODS: Some 54 videoendoscopic adrenalectomies performed on 52 patients by five surgical teams between October 1993 and December 1996 were prospectively evaluated. RESULTS: Indications for endoscopic adrenalectomy were pheochromocytoma (n = 17), primary hyperaldosteronism (n = 15), Cushing's adenoma or disease (n = 7), nonsecreting adenoma (n = 7), single metastasis from adenocarcinoma (n = 2), adenoma with dehydroepiandrostenedione (DHEAS) hypersecretion (n = 3), and ACTH-secreting metastases from a thymoma (n = 1). Of the 54 adrenalectomies performed, 31 were of the left gland, 19 of the right and two bilateral. Laparoscopic adrenalectomy was successful in 50 patients (96%). Median tumor size was 4 cm (range 1.5-12), median operation duration was 80 min (range 59-360), and median postoperative stay was 4 days (range 2-13). One patient required blood transfusion. CONCLUSIONS: Endoscopic adrenalectomy can safely be performed-even sporadically-by surgeons well versed in adrenalectomy techniques for endocrine disorders and trained in endoscopic surgery. PMID- 9348375 TI - Bedside percutaneous endoscopic gastrostomy. A safe alternative for early nutritional support in critically ill trauma patients. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) is a good alternative that provides long-term nutritional support and is associated with minimal morbidity. METHODS: During a 24-month period, we studied 54 critically injured patients who underwent early PEG to provide enteral nutritional support. Patients were selected due to the inability to tolerate intake by mouth secondary to multiple associated injuries, especially to the central nervous system. RESULTS: All patients sustained multiple injuries with an average Injury Severity Score of 27. The mean Glasgow Coma Scale at the time of admission was 7 and at the time of the PEG was 10. Eleven patients (20%) had an intracranial pressure (ICP) device, and there was no significant increase in the mean ICP before, during, or after the procedure. In 63% of patients, tube feedings were interrupted for a variety of problems in the 72 h preceding the PEG, and in 70% of patients an average of five radiographs were obtained to document tube position. In 95% of patients, the nutritional goal was achieved within 48 h of PEG placement. There were one immediate and two delayed complications after PEG placement. There were two deaths, neither related to the PEG placement. CONCLUSIONS: Early PEG in critically injured patients is a safe and effective method of providing access to the GI tract for nutritional support. In patients with significant brain injuries, adequate sedation and the presence of an ICP monitor help to minimize secondary insults to the brain. PMID- 9348376 TI - Laparoscopy in the critically ill. AB - BACKGROUND: Laparoscopy was evaluated in critically ill patients with suspected acute cholecystitis, mesenteric ischemia, or gastrointestinal perforation. We studied laparoscopy to assess its utility, accuracy, and effect on cardiopulmonary stability. METHODS: Twenty-six surgical ICU patients with possible abdominal sepsis underwent laparoscopy. Nineteen were post cardiac surgery; the remainder had other diagnoses. Video laparoscopy was performed with hemodynamic monitoring and inotropic support as needed. Eight patients had bedside laparoscopy. RESULTS: Fifteen patients had suspected acute cholecystitis. Laparoscopy was positive in 10; four had open cholecystectomy, four laparoscopic cholecystectomy, and two tube cholecystostomy. Nine patients had suspected mesenteric ischemia; laparoscopy was positive in five, revealing cirrhosis in two and ischemic bowel in three. Two patients had suspected perforated viscus with colonic perforation in one and one false negative. There were no adverse hemodynamic events. CONCLUSIONS: Laparoscopy can be performed safely in critically ill patients. It is useful in patients with acute cholecystitis and in patients who are post cardiac surgery with refractory lactic acidosis in whom a diagnosis of mesenteric ischemia is considered. PMID- 9348377 TI - Production and systemic absorption of toxic byproducts of tissue combustion during laparoscopic surgery. AB - BACKGROUND: Among the potential hazards of laparoscopic surgery using electrocautery is the intraperitoneal release and subsequent absorption of byproducts of tissue combustion. In a porcine model of laparoscopic surgery with smoke production, our aims were to assess (1) the relationship between levels of intraperitoneal carbon monoxide (CO) and systemic carboxyhemoglobin (COHb) and methemoglobin (MetHb), and (2) intraperitoneal concentrations of other noxious gases, including hydrogen cyanide (HCN), acrylonitrile (Acr), and benzene (Bzn). METHODS: Seven pigs underwent laparoscopic resection of three hepatic wedges using monopolar electrocautery in a CO2 pneumoperitoneum. Sequential arterial samples were drawn to measure [COHb] and [MetHb] perioperatively, while gaseous intraabdominal [CO], [HCN], [Acr], and [Bzn] were assayed intraoperatively. RESULTS: The mean +/- SEM duration of operation was 90 +/- 2 min, and electrocautery was used for 68 +/- 4 min. Intraabdominal [CO] rose from 0 to 814 +/- 200 ppm (p < 0.01) while [COHb] increased from 2.9 +/- 0.1% to 3.5 +/- 0.1% (p < 0.001). Systemic [MetHb] remained unchanged intra- and postoperatively, ranging from 0.3 to 0.7%. Intraperitoneal [HCN] rose from 0 to 5.7 +/- 0.7 ppm (p < 0.001). [Acr], however, did not change significantly from preoperative values, ranging from 0 to 1.6 +/- 1. 0 ppm, and [Bzn] was undetectable. CONCLUSIONS: Laparoscopic tissue combustion increases intraabdominal [CO] to "hazardous" levels leading to minimal, yet significant, elevations of [COHb]. Systemic [MetHb] and intraabdominal [HCN], [Acr], and [Bzn] are not elevated to toxic levels. Production of intraperitoneal smoke during laparoscopic electrosurgery therefore may not pose a significant threat to the patient. PMID- 9348378 TI - Evaluation of laparoscopic Toupet fundoplication as a primary repair for all patients with medically resistant gastroesophageal reflux. AB - BACKGROUND: This prospective study assesses the outcome results in 100 consecutive patients with gastroesophageal reflux disease (GERD) treated with a laparoscopic Toupet fundoplication. METHODS: GERD was confirmed by 24-h pH study and/or esophagogastroduodenoscopy (EGD). Pre- and postoperative symptoms, operative times, and perioperative complications were recorded on standardized data forms. Early follow-up was at 3 months and late follow-up, including 24-h pH, manometry, and EGD was at 22 months. RESULTS: Preoperative symptoms included heartburn (92%), regurgitation (58%), water brash (39%), and dysphagia (39%). Mean operative time was 3.2 hours. There were no conversions to celiotomy and there were no mortalities. The perioperative complication rate was 14%; 6% (5/83) of patients reported heartburn at 3 months and 20% (15/74) at 22 months. Early and late dysphagia was 20% (17/83) and 9% (7/74), respectively; 24-h pH testing was abnormal in 90% of symptomatic patients (9/10), 39% of asymptomatic patients (12/31), and 51% overall. CONCLUSIONS: Despite early improvement in reflux symptoms following laparoscopic Toupet fundoplications, there is a high incidence of recurrent GERD. Symptomatic follow-up underestimates the true incidence of 24 h pH-documented reflux. Based on these results we cannot recommend the laparoscopic Toupet repair for GERD patients with normal esophageal motility. PMID- 9348379 TI - Minimally invasive management of low-grade and benign gastric tumors. AB - BACKGROUND: Benign gastric tumors and tumors of low-grade malignancy can be safely removed laparoscopically. METHODS: Seven patients were considered candidates for laparoscopic resection of gastric tumors. Inclusion criteria included small tumor size (less than 6 cm), exophytic or endophytic tumor morphology, and benign characteristics. Indications for surgical intervention included bleeding, weight loss, and need for tissue diagnosis. Patients ranged in age from 38 to 70. There were five female and two male patients. All patients underwent preoperative upper GI endoscopy. The procedures were performed using a four- or five-port technique. An Endo-GIA (US Surgical Company, Norwalk, Connecticut) was used to amputate those tumors located on the serosal surface of the stomach. Tumors on the mucosal surface were exposed via a gastrotomy, then likewise amputated using an Endo-GIA. The gastrotomy closure was then either hand sewn or stapled. Operating time ranged from 95 to 225 min. RESULTS: Final pathologic diagnoses included lipoma, lymphoma, leiomyoma, and leiomyosarcoma. There was a 28% conversion rate. There were no complications. Length of postoperative stay ranged from 4 to 7 days. There have been no tumor recurrences in 6-38-month follow-up. CONCLUSIONS: Minimally invasive management of benign and low-grade gastric tumors can be performed safely with excellent short- and long term results. PMID- 9348380 TI - Laparoscopic fundoplication to enhance pulmonary function in children with severe reactive airway disease and gastroesophageal reflux disease. AB - BACKGROUND: The relationship between severe reactive airway disease (RAD) and gastroesophageal reflux disease (GERD) has been noted but the relationship is poorly understood. This study reports our experience with laparoscopic fundoplication and its effect on the pulmonary status of children with severe steroid-dependent reactive airway disease. METHODS: Fifty-six patients with severe steroid-dependent RAD and medically refractory GERD underwent laparoscopic Nissen fundoplications. Mean age was 7 years and mean weight was 20 kg. All patients had the procedure completed successfully laparoscopically with an average operative time of 62 min. Average hospital stay was 1.6 days. RESULTS: Forty-eight of 56 patients noted significant improvement in their respiratory symptoms in the first week. Fifty of 56 patients have been weaned off their oral steroids and four others have had a greater than 50% decrease in their dose. Sixteen patients had a documented increase in their FEV1 in the initial postoperative period (avg. 26%). CONCLUSION: Patients with steroid-dependent RAD and GERD refractory to medical management show improvement in their respiratory status following fundoplication and the majority can be weaned off of their oral steroids. Laparoscopic techniques allow this procedure to be performed safely even in this high-risk group of patients. PMID- 9348381 TI - Initial results of a prospective trial of outpatient laparoscopic cholecystectomy. AB - BACKGROUND: Whether or not laparoscopic cholecystectomy may be performed safely as an outpatient procedure is controversial. In 1993, a protocol for outpatient laparoscopic cholecystectomy was instituted to determine the benefits and safety of discharging patients within several hours of surgery. METHODS: The initial 60 outpatient laparoscopic cholecystectomies performed by one surgeon in a hospital based outpatient teaching facility between February 1993 to June 1996 were prospectively studied. RESULTS: Fifty-eight (97%) patients were discharged successfully after an average stay in the recovery room of 3 h. There were no deaths. Two patients required overnight observation and three patients required readmission. Two patients (3%) had cystic duct leak. The average hospital stay for all patients undergoing laparoscopic cholecystectomy at the institution (inpatient and outpatient) decreased from 3.2 to 1.5 days and the average hospital cost decreased from $7,800 to $4,600 during this period. CONCLUSION: Laparoscopic cholecystectomy in an outpatient setting is safe and cost-effective in healthy patients. PMID- 9348382 TI - Final score in laparoscopic cholecystectomy. Cholangiogram 1207, no cholangiogram 116. AB - BACKGROUND: The role of intraoperative fluorocholangiography (IOC) in laparoscopic cholecystectomy (LC) is controversial. We evaluated the use of IOC at an institution where the study is performed routinely. METHODS: Records of all patients undergoing LC during a 3-year period ending January 1, 1996 were reviewed. RESULTS: A total of 1207 patients received IOC, whereas 116 patients did not. IOC findings were categorized as follows: normal, 1016 cases (84%); CBD stone, 149 cases (12.3%); anomalies, 23 cases (1.9%); duodenal diverticula, 10 cases (0.8%); ductal strictures, four cases (0.3%); and CBD diverticula, 5 cases (0.4%). In the 116 patients who did not receive IOC, 35 of the procedures could not be performed, whereas 81 were not attempted. Of the 149 IOC that showed CBD stones, two were false positives. Anomalies included accessory right hepatic ducts (11 cases), cystic ducts joining the right hepatic duct (seven cases), and abnormal cystic duct entries (five cases). Duct injuries occurred in 5 cases (0.4%), three before and two after IOC. Four injuries were minor; IOC prevented CBD transection. CONCLUSIONS: Routine IOC is feasible, safe, accurate, and provides critical information of immediate use during LC. By treating ductal stones at operation and identifying patients without CBD stones, IOC minimizes need for postoperative studies, including endoscopic retrograde cholangiography (ERC). PMID- 9348383 TI - Utility of transesophageal echocardiography and pulmonary artery catheterization during laparoscopic assisted abdominal aortic aneurysm repair. AB - BACKGROUND: Advanced laparoscopic procedures are more commonly performed in elderly patients with cardiac disease. There has been limited data on the use of pulmonary artery catheters (PAC) and transesophageal echocardiography (TEE) to monitor hemodynamic changes. METHODS: We prospectively studied eight patients undergoing laparoscopic assisted abdominal aortic aneurysm repair. All patients had a PAC and all but one had an intraoperative TEE. Data included heart rate (HR), temperature (temp), pulmonary artery systolic (PAS) and diastolic (PAD) pressures, mean arterial pressure (MAP), central venous pressure (CVP), pulmonary capillary wedge pressure (PCWP), cardiac index (CI), mixed venous oxygen saturation (MVO2), and oxygen extraction ratio (O2Ex) and was obtained prior to induction, during insufflation, after desufflation, during aortic cross-clamp, and at the end of the procedure. End diastolic area (EDA), a reflection of volume status, was measured on TEE. ANOVA was used for data analysis. RESULTS: No changes were noted in HR, temp, PAS, PCWP, CI, MVO2, and O2Ex. PAD and CVP were greater during insufflation compared with baseline and aortic cross-clamp without associated changes in EDA. MAP was higher at baseline compared with all other times during the procedure. CONCLUSIONS: Insufflation increased PAD and CVP. However, volume status as suggested by EDA and PCWP did not change. These data question the reliability of hemodynamic measurements obtained from the PAC during pneumoperitoneum and suggest that TEE may be sufficient for evaluation of volume status along with the added benefit of timely detection of ventricular wall motion abnormalities. PMID- 9348384 TI - Mesh configurations in laparoscopic extraperitoneal herniorrhaphy. A comparison of techniques. AB - BACKGROUND: Laparoscopic total extraperitoneal (TEP) hernia repair utilizes slit mesh that is placed around the spermatic cord to secure the prosthesis and prevent recurrence. Because of concern that encircling of the cord might increase pain and morbidity, we compared patients with mesh repairs using encircled and nonencircled techniques. METHODS: The 191 male patients who underwent bilateral TEP repairs were divided into three groups. In 100 consecutive patients (group A), the slit mesh was closed around both spermatic cords; in 56 patients (group B), the slit mesh was tucked under the spermatic cords but not closed; in 35 consecutive patients (group C), the slit was closed around one cord and tucked under the other, in a randomized fashion. RESULTS: The groups had similar operative times (A: 83 +/- 25 min; B: 79 +/- 21; C; 77 +/- 24), use of pain medication (A: 2.7 +/- 2.5 days; B: 2.4 +/- 1.9; C: 3.1 +/- 2.4), and recovery before return to work (A: 7.9 +/- 7.0 days; B: 8.2 +/- 6.1; C: 6.7 +/- 4.8). The incidence of indirect hernias was similar in all groups. Complication rate was 20% in A, 20% in B, and 14% in C (p = NS). Chronic pain was more frequent in A (A: 6, B: 0, p = 0. 06). In group C, fluid collections were more common on the closed side (closed: 3, tucked: 0; p = 0.08). There were no recurrences in any group. CONCLUSIONS: Closing the slit around the spermatic cord in laparoscopic inguinal hernia repair is not essential for prevention of early recurrence. Fluid collections tended to be more frequent when the mesh was closed around the cord, and chronic pain was more frequent in the group with closed mesh bilaterally. PMID- 9348385 TI - Transcystic biliary decompression after direct laparoscopic exploration of the common bile duct. AB - BACKGROUND: A purpose-designed transcystic common bile duct (CBD) decompression cannula is described for use as an alternative to T-tube insertion following laparoscopic direct CBD exploration. This permits safe primary closure of the choledochotomy. METHODS: Following direct supraduodenal laparoscopic clearance of large common bile duct stones, the biliary decompression cannula is inserted percutaneously inside its peel-away sheet over a guide-wire into the CBD via the cystic duct. When in place, the cannula is secured to the cystic duct by two catgut extracorporeal Roeder knots and the choledochotomy is then closed. The terminal multiperforated S-shaped segment of the Cuschieri biliary decompression cannula prevents postoperative dislodgement. RESULTS: Transcystic decompression of the extrahepatic biliary tract using the Cuschieri cannula has been used in 12 patients who underwent laparoscopic supraduodenal CBD exploration for large or occluding stones. There was no instance of postoperative dislodgement of the cannula and all patients had effective drainage of the common bile duct (average 300 ml bile per 24 h). The procedure was uncomplicated in all but one patient who developed self-limiting leakage from the CBD suture line in the early postoperative period. The median hospital stay after surgery was 4 days, with a range of 3 to 10 days. The cystic duct decompression cannula was capped and sealed under an occlusive dressing at the time of discharge. Removal of the cannula was carried out without any complications as a day case 11-16 days after surgery. CONCLUSIONS: Transcystic biliary decompression is safe and effective. The experience with is use indicates that compared to T-tube drainage, transcystic decompression may accelerate recovery and reduce the hospital stay in patients following laparoscopic direct exploration of the CBD. Its insertion is less technically demanding than placing a T-tube through the choledochotomy. Transcystic decompression with complete primary closure of the CBD realizes the full benefits of the single-stage management of common bile duct calculi and permits confirmation of complete stone clearance after surgery. PMID- 9348386 TI - Laparoscopic colposuspension using mesh reinforcement. AB - BACKGROUND: For patients with stress urinary incontinence, surgical reestablishment of the bladder neck has proved amenable to a laparoscopic approach, which shortens hospitalization and reduces tissue trauma. The use of mesh reinforcement to improve the durability of colposuspension can refine this proven procedure even further. METHODS: We performed laparoscopic Burch colposuspension on 54 patients with stress urinary incontinence and compared our results with those of other investigators. RESULTS: All patients reported resolution of incontinence postoperatively: 83.3% received no supplementary medication while 16.7% took antispasmodic-anticholinergic medications. Two cases required conversion to an open procedure. Hospital stay declined from 2.7 days (first quartile) to 1.9 days (last quartile) (average, 2.3 days). Complications were rare, and in a 28-month follow-up, no reoperations were required. CONCLUSION: Laparoscopic Burch colposuspension using mesh reinforcement provides durable resolution of stress incontinence with low risk of conversion, short hospitalization, and few complications. PMID- 9348387 TI - Laparoscopic cryosurgery for hepatic tumors. Experimental observations and a case report. AB - BACKGROUND: Hepatic cryosurgery has been shown to be a safe technique that may be well suited to a laparoscopic approach. METHODS: The technical feasibility and safety of laparoscopic cryosurgery was explored first in a pig model. Thereafter we performed the first successful case of laparoscopic hepatic cryosurgery at our institution. RESULTS: In the animal model, we found that it is possible to safely identify, target, and cryoablate specific lesions in the liver. Temperature in the peritoneal cavity remained above 35 degrees C, and pathologic examination of the abdominal wall around the cryoprobe site revealed no damage. We also successfully treated a 62-year-old man with a metastatic colorectal carcinoma deep in the right lobe of the liver with laparoscopic cryosurgery using a transpleural approach. CONCLUSION: We conclude that laparoscopic cryosurgery is feasible for lesions anywhere in the liver. For lesions high on the dome of the liver, a transpleural approach may provide better access. PMID- 9348388 TI - Laparoscopically assisted radical sacrococcygectomy. A new operative approach to large sacrococcygeal chordomas. AB - Laparoscopic rectal mobilization and hypogastric arterial isolation were combined with a posterior sacrococcygectomy for the resection of a large sacrococcygeal chordoma in two patients. The laparoscopic procedure as described was uneventful in both cases. There was no postoperative morbidity associated with the laparoscopic procedure. The combination of laparoscopic pelvic dissection and radical posterior sacrococcygectomy is safe, effective, oncologically sound, and should be considered for all patients with a large proximal sacrococcygeal chordoma. PMID- 9348389 TI - Endoscopic nasobiliary drainage for bile duct injury after laparoscopic cholecystectomy. AB - Bile duct injuries are a potential complication of laparoscopic cholecystectomy (LC). A patient who underwent successful endoscopic nasobiliary drainage (ENBD) for a bile duct injury sustained during LC is presented. Of particular note, the patient also had Chilaiditi's syndrome. A 59-year-old woman was admitted with symptomatic cholecystolithiasis and Chilaiditi's syndrome. LC was performed. Postoperatively, the patient complained of abdominal discomfort. Laboratory examination revealed cholestasis. Bilious material began spilling from an intraabdominal drain. Subsequent endoscopic retrograde cholangiopancreatography (ERCP) showed bile leakage. ENBD was performed. Repeat ERCP 10 days later failed to show a bile leak or stenosis of the common bile duct. The patient improved rapidly and had no complaints after the procedure. ENBD is a useful endoscopic technique to prevent peritonitis from bile leakage after LC. Chilaiditi's syndrome is not a contraindication for LC. PMID- 9348390 TI - Intraoperative ultrasound assessment in management of complex pancreatic pseudocysts. AB - Preoperative imaging studies and operative inspection may provide insufficient information to appropriately manage certain complex pancreatic pseudocysts. Intraoperative ultrasound accurately identifies and localizes peripancreatic fluid collections, cyst wall thickness, parenchymal and ductal anatomy, and relationships to adjacent visceral and vascular structures. Adjunctive use of intraoperative ultrasonography altered the surgical management in the clinical case described herein and is advocated for assessment of problematic pancreatic pseudocysts. PMID- 9348391 TI - Laparoscopic management of lumbar hernia. AB - We describe (for the first time) a laparoscopic approach to repair an acquired superior triangle lumbar hernia in a morbidly obese woman by using prosthetic mesh. Such a technique provides an excellent anatomic view, thus avoiding injury to structures in proximity to the hernia during repair; eventually the well-known advantages of such approach result. PMID- 9348392 TI - A simple and useful method for retracting the left liver lobe. PMID- 9348393 TI - Splenic rupture from colonoscopy. PMID- 9348394 TI - New endoscopic methods for the detection of early colorectal cancer. What are we doing? PMID- 9348395 TI - Endoscopic thyroid and parathyroid surgery. PMID- 9348396 TI - Importance of transesophageal echocardiography in directing the surgical approach to atrial myxomas. PMID- 9348397 TI - Morphology and structure of polytene chromosomes. PMID- 9348398 TI - Basal ganglia pathophysiology. A critical review. PMID- 9348399 TI - The striatopallidal fiber system in primates. PMID- 9348400 TI - The subthalamic nucleus and its connections. New electrophysiological and pharmacological data. PMID- 9348401 TI - Spatial relationships between striatal axonal endings and pallidal neurons in macaque monkeys. PMID- 9348402 TI - New concepts about the organization of basal ganglia output. PMID- 9348404 TI - Anatomical segregation of information processing in the rat substantia nigra pars reticulata. PMID- 9348405 TI - The pedunculopontine nucleus and related structures. Functional organization. PMID- 9348403 TI - New data on the anatomical organization of the thalamostriatal projections. PMID- 9348406 TI - Neuronal activity in the basal ganglia. Functional implications. PMID- 9348407 TI - Neurochemical correlates of parkinsonism. Role of dopaminergic lesions. PMID- 9348408 TI - Basal ganglia function and dysfunction revealed by PET activation studies. PMID- 9348409 TI - An introduction to the new surgery for Parkinson's disease. Past and present problems. PMID- 9348410 TI - Influence of clinical presentation on target selection. PMID- 9348411 TI - Microelectrode recording and stimulation techniques during stereotactic procedures in the thalamus and pallidum. PMID- 9348412 TI - Microelectrode recording-guided posteroventral pallidotomy in patients with Parkinson's disease. PMID- 9348413 TI - Electrophysiological targeting in stereotaxic surgery for Parkinson's disease. PMID- 9348414 TI - Microelectrode-guided pallidotomy for medically intractable Parkinson's disease. PMID- 9348415 TI - Medial pallidotomy in late-stage Parkinson's disease and striatonigral degeneration. PMID- 9348416 TI - Chronic intracerebral stimulation in Parkinson's disease. PMID- 9348417 TI - Thalamic surgery for movement disorders. PMID- 9348418 TI - The role of the subthalamic nucleus in the origin of hemiballism and parkinsonism: new surgical perspectives. PMID- 9348419 TI - Neural transplantation as a therapy for Parkinson's disease. PMID- 9348420 TI - PET modifications after surgery for Parkinson's disease. PMID- 9348421 TI - Early detection of Alzheimer disease: methods, markers, and misgivings. AB - There is at present no reliable predictive test for most forms of Alzheimer disease (AD). Although some information about future risk for disease is available in theory through ApoE genotyping, it is of limited accuracy and utility. Once neuroprotective treatments are available for AD, reliable early detection will become a key component of the treatment strategy. We recently conducted a pilot survey eliciting attitudes and beliefs toward an unspecified and hypothetical predictive test for AD. The survey was completed by a convenience sample of 176 individuals, aged 22-77, which was 75% female, 30% African-American, and of which 33% had a family member with AD. The survey revealed that 69% of this sample would elect to obtain predictive testing for AD if the test were 100% accurate. Individuals were more likely to desire predictive testing if they had an a priori belief that they would develop AD (p = 0.0001), had a lower educational level (p = 0.003), were worried that they would develop AD (p = 0.02), had a self-defined history of depression (p = 0.04), and had a family member with AD (p = 0.04). However, the desire for predictive testing was not significantly associated with age, gender, ethnicity, or income. The desire to obtain predictive testing for AD decreased as the assumed accuracy of the hypothetical test decreased. A better short-term strategy for early detection of AD may be computer-based neuropsychological screening of at-risk (older aged) individuals to identify very early cognitive impairment. Individuals identified in this manner could be referred for diagnostic evaluation and early cases of AD could be identified and treated. A new self-administered, touch-screen, computer based, neuropsychological screening instrument called Neurobehavioral Evaluation System-3 is described, which may facilitate this type of screening. PMID- 9348422 TI - Slowing the progression of Alzheimer disease: monitoring progression. AB - The search for biologic markers of Alzheimer disease (AD) progression has thus far had limited success. A technique is described, involving magnetic resonance imaging, that can be used to follow the progress of brain atrophy and hence the loss of neuronal tissue. It involves acquisition of three-dimensional images of the brain on two consecutive occasions and their precise co-registration so as to assess the loss of tissue volume over time. Median loss of brain volume in a group of individuals with sporadic and familial AD ranged from about 5-20 ml/year compared to less than 2 ml/year for controls. Loss of cerebral tissue volume was also seen in two individuals from a pedigree with familial AD at a time when symptoms were only just apparent. PMID- 9348423 TI - Slowing the progression of Alzheimer disease: methodologic issues. AB - The evidence to support a claim that a new drug will slow the progression of Alzheimer disease (AD) must derive from epistemologically valid research methods. Although agency regulations do not specify the magnitude of an effect that a drug must possess to be granted a claim as a treatment for AD, the evidence to support any claim must be adduced in adequate and well-controlled clinical investigations and must meet the standard of "substantial evidence." Because a claim presented in drug product labeling may not be false or misleading in any particular, a distinction must be made between treatments that provide a "symptomatic" benefit and those that alter the course of dementia. Examples of some of the difficulties likely to be encountered by sponsors seeking to develop evidence to support a claim that a new drug slows the progression of dementia are presented. A suggestion is made for a clinical trial design, designated as the "randomized start design," that may be useful in such a question. Why this design might overcome many of the difficulties, both practical and ethical, present in the "discontinuation" design, the design ordinarily proposed to assess a drug's effect on disease progression, is discussed. PMID- 9348424 TI - Pharmacoeconomics of dementia. AB - Pharmacoeconomics is a new discipline with its own terminology and techniques, little known to most clinicians but destined to become more important to them and to their patients. This article reviews basic approaches in pharmacoeconomics; current knowledge of costs related to dementia; assessment of quality of life in patients with cognitive impairment; models of relationships among degree of cognitive impairment, caregiver burden, and health care utilization; existing pharmacoeconomic guidelines for drug studies; pharmacoeconomic studies of tacrine, a drug used in Alzheimer disease; and further work needed to advance the critical area of pharmacoeconomic studies in dementia. PMID- 9348425 TI - Slowing the progression of Alzheimer disease: ethical issues. AB - Slowing the progression of Alzheimer disease (AD) can be regarded as an unambiguous benefit only up to a point. Beyond that point, the greater human good will involve letting the downward progression proceed. However, defining this point is not easy. It may be useful to distinguish the then-self, i.e., the intact self that relates past, present, and future, from the now-self, i.e., a state in which the demented individual recognizes only the present. In decisions involving treatment and the prolongation of life, many believe that the (precedently expressed wishes of the) then-self takes precedence. Decisions about slowing disease progression involve not only the then-self but the medical community and the caregivers. They, too, may agree that slowing the progression of AD beyond a critical point is unacceptable. PMID- 9348426 TI - Isolation and chemical-structural determination of a novel aromatic amine mutagen in water from the Nishitakase River in Kyoto. AB - Water samples from the Nishitakase River in Kyoto, Japan, especially taken at sites below sewage plants, show significantly high mutagenicity in the Ames test. In the present study, mutagens in the river water were adsorbed to 24 g of blue rayon, extracted, and separated by HPLC on ODS columns. Five mutagenic compounds (I-V) were isolated, and they accounted for 21%, 17%, 11%, 12%, and 6%, respectively, of the total mutagenicity of the blue rayon-adsorbed materials. With compound I obtained from adsorbate to 24 g of blue rayon as a marker, a large quantity (1.1 mg) of mutagenic compound I was isolated by Sephadex LH-20 column chromatography and HPLC on ODS columns from material adsorbed to 27 kg of blue cotton. X-ray crystal analysis was carried out with the debrominated derivative of compound I. Based on this X-ray crystallography data and the UV, mass, and 1H-NMR spectra of both the derivative and compound I, the structure of compound I was determined to be 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5 methoxyphenyl]-5-amino - 7-bromo-4-chloro-2H-benzotriazole (PBTA-1). PBTA-1 is a newly identified potent mutagen, inducing 1,200,000 revertants of Salmonella typhimurium YG1024 per microgram in the presence of S9 mix. PMID- 9348427 TI - Solid phase microextraction as a tool to determine membrane/water partition coefficients and bioavailable concentrations in in vitro systems. AB - Solid phase microextraction (SPME) is an extraction technique that uses a polymer coated fiber as the extraction device. After extraction, the compound of interest can be desorbed from the fiber and subsequently analyzed by GC or HPLC. One of the properties of SPME is that only the freely dissolved fraction of a chemical is available for partitioning to the extraction device. The method can be applied in a way that small amounts are extracted from the sample, which allows negligible depletion extraction. These two properties make SPME devices particularly suitable for measurements of free concentrations. In toxicological studies the free concentration is considered to be a more relevant parameter, concerning toxic effects, than the nominal concentration that is used most frequently. In the current study, the usefulness of this method to measure phospholipid/water partition coefficients and free concentrations in three different in vitro test systems (rat hepatocytes in primary culture, 9000 g and 100,000 g homogenate fractions of rainbow trout liver) was demonstrated. Results show separate relationships between phospholipid/water and n-octanol/water partition coefficients for a set of polar and nonpolar organic chemicals, respectively. These observations suggest that phospholipid/water partition coefficients may be a more suitable parameter in modeling the kinetic behavior of organic chemicals. Additionally, differences between the nominal and the actual free concentration in in vitro systems are more pronounced for more hydrophobic compounds, as was expected based on theoretical considerations. To our knowledge, the approach presented here is the first analytical method to measure toxicologically relevant concentrations in in vitro test systems in a fast and efficient way. PMID- 9348428 TI - Regioselective hydroxylations of 2-nitrofluorene in vivo: a nuclear magnetic resonance study. AB - The time-dependent metabolism of 2-nitrofluorene (2-NF) in vivo has been studied after ip administration to rats. After hydrolysis with beta glucuronidase/arylsulfatase, five hydroxylated 2-nitrofluorenes were isolated by HPLC and identified by a combination of data from NMR, UV, and MS experiments. These metabolites were characterized as 6,9-dihydroxy-2-nitrofluorene (I), 9 hydroxy-2-nitrofluorene (II), 6-hydroxy-2-nitrofluorene (III), 7-hydroxy-2 nitrofluorene (IV), and 8-hydroxy-2-nitrofluorene (V). The highest amounts of metabolites were found after 24 h of injection, and although the major ones were still present after 48 h, only traces of them could be found beyond that time. A unique feature of the present study was our ability to establish the position of the different hydroxylations that occur in vivo by NMR techniques and, thus, to observe the regioselectivity of the metabolism of the 2-NF in vivo. Furthermore, two conjugated metabolites were identified by 1H-NMR and ESI-MS as 6 [(hydroxysulfonyl)oxy]-2-nitrofluorene (2-nitrofluorene 6-sulfate) (VI) and 7 [(hydroxysulfonyl)oxy]-2-nitrofluorene (2-nitrofluorene 7-sulfate) (VII). PMID- 9348429 TI - Intraphagosomal chlorination dynamics and yields determined using unique fluorescent bacterial mimics. AB - Fluorescein was covalently attached through a cystamine linker group to carboxy derivatized polyacrylamide microspheres to generate phagocytosable particles containing fluorescent reporter groups. A unique feature of these beads is that the dye was recoverable in near-quantitative yield from intracellular environments by thiol reduction of the cystamine disulfide bond. Fluorescence microscopy indicated that individual neutrophils could bind as many as approximately 20 serum-opsonized beads, although no appreciable cellular association was observed for unopsonized beads. By using methyl viologen to quench external fluorescence, it was demonstrated that 70-90% of the neutrophil associated fluorescein on opsonized beads was inaccessible to the medium. The particle-bound fluorescein underwent near-stoichiometric conversion to chlorinated derivatives when reacted with HOCl or the cell-free myeloperoxidase (MPO)-H2O2-Cl- system; products were identified by HPLC separation and electrospray ionization mass spectrometry of the recovered dye. Fluorescence changes accompanying phagocytosis were consistent with chlorination of the dye; fluorescence spectrometric and chemical trapping measurements indicated that intraphagosomal chlorination was far more extensive than extracellular chlorination. Yields of recovered chlorofluoresceins determined by HPLC indicated that sufficient HOCl had been produced intracellularly to kill entrapped bacteria. Fluorescein chlorination coincided approximately with phagocytosis and stimulated uptake of O2 by the cells. Demonstration that HOCl is produced within phagosomes in sufficient concentrations to kill bacteria on a time scale associated with death constitutes strong evidence in support of a primary role for HOCl in the microbicidal action of neutrophils. PMID- 9348430 TI - Lipid peroxidation-dependent chemiluminescence from the cyclization of alkylperoxyl radicals to dioxetane radical intermediates. AB - This work reveals a novel mechanism for triplet carbonyl formation (and hence chemiluminescence) during lipid peroxidation, whose chemiluminescence has been attributed to both triplet carbonyls and singlet oxygen. As a model for polyunsaturated fatty acid hydroperoxides, we have synthesized 3-hydroperoxy-2,3 dimethyl-1-butene by photooxygenation of tetramethylethylene. One-electron oxidation of this hydroperoxide with heme proteins and peroxynitrite to the corresponding alkylperoxyl radical results in chemiluminescence, both direct and 9,10-dibromoanthracene-2-sulfonate-sensitized, the latter attributed to the formation of triplet acetone. It is postulated that triplet acetone results from the cyclization of the alkylperoxyl radical to a dioxetane radical intermediate followed by its thermolysis. This is supported by EPR spin-trapping experiments in which discrimination between carbon-centered radicals derived from the alkyloxyl and alkylperoxyl radicals is achieved through the use of one-electron oxidants and reductants, e.g., FeII- and TiIII. PMID- 9348431 TI - Inactivation of glyceraldehyde-3-phosphate dehydrogenase by a reactive metabolite of acetaminophen and mass spectral characterization of an arylated active site peptide. AB - Acetaminophen (4'-hydroxyacetanilide, APAP) is a widely used analgesic and antipyretic drug that can cause hepatic necrosis under some circumstances via cytochrome P450-mediated oxidation to a reactive metabolite, N-acetyl-p benzoquinone imine (NAPQI). Although the mechanism of hepatocellular injury caused by APAP is not fully understood, it is known that NAPQI forms covalent adducts with several hepatocellular proteins. Reported here is the identification of one of these proteins as glyceraldehyde-3-phosphate dehydrogenase [GAPDH, D glyceraldehyde-3-phosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12]. Two hours after the administration of hepatotoxic doses of [14C]APAP to mice, at a time prior to overt cell damage, hepatocellular GAPDH activity was significantly decreased concurrent with the formation of a 14C-labeled GAPDH adduct. A nonhepatotoxic regioisomer of APAP, 3'-hydroxyacetanilide (AMAP), was found to decrease GAPDH activity to a lesser extent than APAP, and radiolabel from [14C]AMAP bound to a lesser extent to GAPDH at a time when its overall binding to hepatocellular proteins was almost equivalent to that of APAP. In order to determine the nature of the covalent adduct between GAPDH and APAP, its major reactive and toxic metabolite, NAPQI, was incubated with purified porcine muscle GAPDH. Microsequencing analysis and fast atom bombardment mass spectrometry (FAB-MS) with collision-induced dissociation (CID) were used to characterize one of the adducts as APAP bound to the cysteinyl sulfhydryl group of Cys-149 in the active site peptide of GAPDH. PMID- 9348432 TI - In vivo evidence of free radical formation in the rat lung after exposure to an emission source air pollution particle. AB - Exposure to air pollution particles can be associated with increased human morbidity and mortality. The mechanism(s) of lung injury remains unknown. We tested the hypothesis that lung exposure to oil fly ash (an emission source air pollution particle) causes in vivo free radical production. Electron spin resonance (ESR) in conjunction with the spin trap alpha-(4-pyridyl 1-oxide)-N tert-butylnitrone (4-POBN) was used to detect radical adducts. Rats were instilled with 500 micrograms of either oil fly ash or saline. Twenty-four hours later, ESR spectroscopy of the chloroform extract from lungs of animals exposed to the oil fly ash gave a spectrum consistent with a carbon-centered radical adduct (hyperfine coupling constants alpha N = 15.0 G and alpha H beta = 2.5 G), while those spectra from lungs instilled with saline revealed a much weaker signal. This signal was reproduced by instilling animals with the soluble fraction of the oil fly ash, which contains soluble metal compounds. The same signal was observed after instillation of either a mixture of vanadium, nickel, and iron sulfates or VOSO4 alone. We conclude that, after instillation of an air pollution particle in the rat, ESR analysis of lung tissue demonstrates in vivo free radical production. This generation of free radicals appears to be associated with soluble metals in the oil fly ash. PMID- 9348433 TI - Immunochemical visualization and identification of rat liver proteins adducted by 2,6-di-tert-butyl-4-methylphenol (BHT). AB - Several alkylphenols (e.g., 2,6-di-tert-butyl-4-methylphenol, BHT) form reactive quinone methide intermediates (e.g., 2,6-di-tert-butyl-4-methylene-2,5 cyclohexadienone, BHT-QM) upon oxidation by cellular enzymes. In order to pursue the role of protein alkylation in alkylphenol toxicity, we used an immunochemical approach to identify protein targets alkylated by BHT. Synthetic BHT-N acetylcysteine (BHT-NAC) was coupled to keyhole limpet hemocyanin and used as an antigen from which polyclonal antibodies were raised in New Zealand white rabbits. Rabbit serum contained an antibody which was highly specific for BHT NAC, as determined by competitive ELISA. The BHT antibody was used as a probe to look for the presence of BHT-protein adducts in in vitro incubations with rat liver microsomes or tissue slices and also in vivo in liver tissue from male Sprague-Dawley rats exposed to BHT. Western blotting of protein gels revealed BHT dependent protein alkylation over a wide molecular weight range. Prominent recurrent bands were observed at approximately 34.5, 52, 64.5, 74, and 97 kDa. Detection of adducts was inhibited in microsomal incubations by cytochrome P450 inhibitors, deuterated BHT, and the omission of NADPH. Similar protein alkylation patterns were observed in rat liver microsomes exposed to synthetically prepared BHT-QM as in the enzyme-mediated incubations. In rats gavaged with up to 1000 mg/kg BHT, the amount of protein alkylation observed was maximal at 24 h postdosing and was dose-dependent. Two alkylated proteins were isolated and identified by N-terminal sequencing: a mitochondrial beta-oxidation enzyme, enoyl CoA hydratase, and a plasma membrane/cytoskeletal linker protein from the ezrin/moesin/radixin family. PMID- 9348434 TI - Differential effects of DNA supercoiling on radical-mediated DNA strand breaks. AB - Supercoiling is an important feature of DNA physiology in vivo. Given the possibility that the reaction of genotoxic molecules with DNA is affected by the alterations in DNA structure and dynamics that accompany superhelical tension, we have investigated the effect of torsional tension on DNA damage produced by five oxidizing agents: gamma-radiation, peroxynitrite, Fe2+/ EDTA/H2O2, Fe2+/H2O2, and Cu2+/H2O2. With positively supercoiled plasmid DNA prepared by a recently developed technique, we compared the quantity of strand breaks produced by the five agents in negatively and positively supercoiled pUC19. It was observed that strand breaks produced by gamma-radiation, peroxynitrite, and Fe2+/EDTA/H2O2 were insensitive to DNA superhelical tension. These results are consistent with a model in which chemicals that generate highly reactive intermediates (e.g., hydroxyl radical), but do not interact directly with DNA, will be relatively insensitive to the changes in DNA structure and dynamics caused by superhelical tension. In the case of Fe2+ and Cu2+, metals that bind to DNA, only Cu2+/H2O2 proved to be sensitive to DNA superhelical tension. Strand breaks produced by Cu2+/H2O2 in the positively supercoiled substrate occurred at lower Cu concentrations than in negatively supercoiled DNA. Furthermore, a sigmoidal Cu2+/H2O2 damage response was observed in the negatively supercoiled substrate but not in positively supercoiled DNA. The results with Cu2+ suggest that the redox activity, DNA binding orientation, or DNA binding affinity of Cu1+ or Cu2+ is sensitive to superhelical tension, while the results with the other oxidizing agents warrant further investigation into the role of supercoiling in base damage. PMID- 9348435 TI - A molecular mechanics and dynamics study of the minor adduct between DNA and the carcinogen 2-(acetylamino)fluorene (dG-N2-AAF). AB - Experimental studies involving the carcinogenic aromatic amine 2 (acetylamino)fluorene (AAF) have afforded two acetylated DNA adducts, the major one bound to C8 of guanine and a minor adduct bound to N2 of guanine. The minor adduct may be important in carcinogenesis because it persists, while the major adduct is rapidly repaired. Primer extension studies of the minor adduct have indicated that it blocks DNA synthesis, with some bypass and misincorporation of adenine opposite the lesion [Shibutani, S., and Grollman, A.P. (1993) Chem. Res. Toxicol. 6, 819-824]. No experimental structural information is available for this adduct. Extensive minimized potential energy searches involving thousands of trials and molecular dynamics simulations were used to study the conformation of this adduct in three sequences: I, d(C1-G2-C3-[AAF]G4-C5-G6-C7).d(G8-C9-G10-C11 G12-C13-G14+ ++); II, the sequence of Shibutani and Grollman, d(C1-T2-A3-[AAF]G4 T5-C6-A7).d(T8-G9-A10-C11-T12-A13-G14); and III, which is the same as II but with a mismatched adenine in position 11, opposite the lesion. AAF was located in the minor groove in the low-energy structures of all sequences. In the lowest energy form of the C3-[AAF]G4-C5 sequence I, the fluorenyl rings point in the 3' direction along the modified strand and the acetyl in the 5' direction. These orientations are reversed in the second lowest energy structure of this sequence, and the energy of this structure is 1.4 kcal/mol higher. Watson Crick hydrogen bonding is intact in both structures. In the two lowest energy structures of the A3-[AAF]G4-T5 sequence II, the AAF is also located in the minor groove with Watson-Crick hydrogen bonding intact. However, in the lowest energy form, the fluorenyl rings point in the 5' direction and the acetyl in the 3' direction. The energy of the structure with opposite orientation is 5.1 kcal/mol higher. In sequence III with adenine mismatched to the modified guanine, the lowest energy form also had the fluorenyl rings oriented 5' in the minor groove with intact Watson-Crick base pairing. However, the mispaired adenine adopts a syn orientation with Hoogsteen pairing to the modified guanine. These results suggest that the orientation of the AAF in the minor groove may be DNA sequence dependent. Mobile aspects of favored structures derived from molecular dynamics simulations with explicit solvent and salt support the essentially undistorting nature of this lesion, which is in harmony with its persistence in mammalian systems. PMID- 9348436 TI - Synthesis of oligonucleotides containing the ethylene dibromide-derived DNA adducts S-[2-(N7-guanyl)ethyl]glutathione, S-[2-(N2-guanyl)ethyl]glutathione, and S-[2-(O6-guanyl)ethyl]glutathione at a single site. AB - The carcinogen ethylene dibromide (EDB) is activated by enzymatic conjugation with GSH to form S-(2-bromoethyl)GSH, which reacts with DNA via an episulfonium ion. The major DNA adduct derived from EDB was previously characterized as S-[2 (N7-guanyl)ethyl]GSH, and S-[2-(N2-guanyl)ethyl]GSH and S-[2-(O6-guanyl)ethyl]GSH are minor adducts [Cmarik, J. L., Humphreys, W. G., Bruner, K. L., Lloyd, R. S., Tibbetts, C., and Guengerich, F. P. (1992) J. Biol. Chem. 267, 6672-6679]. S-[2 (N7-Guanyl)ethyl]GSH has been incorporated at the G* site in d(5'-TGCTG*CAAG-3'), a site previously found to show GC to AT transitions following treatment of M13 phage with S-(2-chloroethyl)GSH, and the desired product was separated by HPLC. This was ligated to d(5'-GGTACCGAG-3') to yield d(5'-TGCTG*CAAGGGTACCGAG-3'). S [2-(N2-Guanyl)ethyl]GSH was incorporated into the G* site of the oligonucleotide in d(5'-TGCTG*CAAGGGTACCGAG-3') by reacting S-(2-aminoethyl)GSH with an oligomer containing 2-fluoro-O6-[(trimethylsilyl)ethoxy]deoxyinosine at the target site. The 5'-(dimethoxytrityl)-N2-(phenoxyacetyl)-N-[(fluorenylmethyl)formyl ] derivative of S-[2-(O6-deoxyguanosyl)-ethyl]GSH dimethyl ester was synthesized by Mitsunobu alkylation of 5'-(dimethoxytrityl)-N2-(phenoxyacetyl)deoxyguanosine with N-[(fluorenylmethyl)formyl]-S-(2-hydroxyethyl)GSH dimethyl ester, modified to form the phosphoramidite derivative, and incorporated at the G* site of d(5' TGCTG*CAAGGGTACCGAG-3'). The protective groups were removed with 0.10 N NaOH to give the modified oligonucleotide containing S-[2-(O6-guanyl)ethyl]GSH. Although the overall yields were low, the synthesis of a single set of target site oligonucleotides containing these overall yields were low, the synthesis of a single set of target site oligonucleotides containing these three known guanyl adducts allows for in vitro site-specific misincorporation studies. PMID- 9348437 TI - Synthesis and characterization of aflatoxin B1 mercapturic acids and their identification in rat urine. AB - Biologic effects of the hepatocarcinogenic mycotoxin aflatoxin B1 are principally induced by one of its metabolites, the exo-aflatoxin B1 epoxide which produces both DNA and protein adducts in vivo. Detoxication of the exo-aflatoxin B1 epoxide can be mediated in part by glutathione S-transferases whose induction could be important in chemoprotection interventions. Thus, biomarkers of the enzymatic conjugation of exo-aflatoxin B1 epoxide with glutathione may be important indices of protection against the toxic effects of this agent. Since glutathione conjugates undergo further metabolic processing in vivo to yield mercapturic acids, increased urinary excretion of exo-aflatoxin B1 mercapturate could be expected during chemoprotection intervention. To determine if this mercapturic acid could be used as a biomarker, techniques for its specific measurement were developed using monoclonal antibody immunoaffinity chromatography and reverse phase high-performance liquid chromatography with ultraviolet absorbance and mass spectral detection. First, a synthetic exo aflatoxin B1 mercapturate was characterized using mass spectrometry, ultraviolet absorbance, circular dichroism spectrometry, and chemical derivatization. In vivo metabolite characterization was then facilitated by comparison with the synthetically prepared exo-aflatoxin B1 mercapturate and both aflatoxin B1 glutathione conjugate diastereoisomers. In rats, 1% of the aflatoxin dose was excreted as exo-aflatoxin B1 mercapturate within 24 h. The finding that exo aflatoxin B1 mercapturate was excreted in urine in a dose-dependent manner provides the basis for investigating its applicability as a biomarker of glutathione S-transferase status in aflatoxin chemoprotection studies. PMID- 9348438 TI - Urinary metabolites from F344 rats and B6C3F1 mice coadministered acrylamide and acrylonitrile for 1 or 5 days. AB - The purpose of this study was to examine the feasibility of using 13C NMR spectroscopy to analyze urinary metabolites produced following coadministration of two structurally similar carbon-13-labeled compounds to rodents. Acrylonitrile (AN) and acrylamide (AM) are used in the chemical industry to manufacture plastics and polymers. These compounds are known to produce carcinogenic, reproductive, or neurotoxic effects in laboratory animals. The potential for human exposure to AN and AM occurs in manufacturing facilities and environmentally. Male F344 rats and B6C3F1 mice were coadministered po [1,2,3 13C]AN (16-17 mg/kg) and [1,2,3-13C]AM (21-22 mg/kg) after 0 or 4 days of administration of unlabeled AN or AM. Urine was collected for 24 h following administration of the 13C-labeled compounds and analyzed by 13C NMR spectroscopy. Rats and mice excreted metabolites derived from glutathione (GSH) conjugation with AM or AN or derived from GSH conjugation with the epoxides cyanoethylene oxide (CEO) or glycidamide (GA). GA and its hydrolysis product were also detected in the urine of rats and mice. For mice, an increased urinary excretion of total AN- and total AM-derived metabolites (p < 0.05) on repeated coadministration suggested a possible increase in metabolism via oxidation. In addition, mice had an increased (p < 0.05) percentage of dose excreted as metabolites derived from GSH conjugation with AM, AN, CEO, or GA after five exposures as compared with one exposure that may be related to a significant increase in the synthesis of GSH or an increase in glutathione transferase activity. The only significant (p < 0.05) increase between one and five exposures for the rat was in the percentage of metabolites produced following conversion of AM to GA. The use of 13C NMR spectroscopy has provided a powerful methodology for elucidation of the metabolism of two 13C-labeled chemicals administered simultaneously. PMID- 9348439 TI - Metabolic activation of the (+)-S,S- and (-)-R,R-enantiomers of trans-11,12 dihydroxy-11,12-dihydrodibenzo[a,l]pyrene: stereoselectivity, DNA adduct formation, and mutagenicity in Chinese hamster V79 cells. AB - Polycyclic aromatic hydrocarbons require metabolic activation in order to exert their biological activity initiated by DNA binding. The metabolic pathway leading to bay or fjord region dihydrodiol epoxides as ultimate mutagenic and/or carcinogenic metabolites is thought to play a dominant role. For dibenzo[a,l]pyrene, considered as the most potent carcinogenic polycyclic aromatic hydrocarbon, the formation of the fjord region syn- and/or anti-11,12 dihydrodiol 13,-14-epoxide (DB[a,l]PDE) diastereomers has been found to be the principal metabolic activation pathway in cell cultures leading to DNA adducts. In order to further elucidate the stereoselectivity involved in this activation pathway via the formation of the trans-11,12-dihydrodiol, we have synthesized the enantiomerically pure 11,12-dihydrodiols of dibenzo[a,l]-pyrene and investigated their biotransformation in rodents. Incubations with liver microsomes of Sprague Dawley rats and CD-1 mice pretreated with Aroclor 1254 revealed that the enzymatic conversion to the fjord region DB[a,l]PDE strongly depends on the absolute configuration of the 11,12-dihydrodiol enantiomers. While oxidation at the 13,14-position of the (+)-(11S,12S)-dihydrodiol is limited to a small extent, the (-)-11R,12R-enantiomer is metabolized to its fjord region dihydrodiol epoxides in considerably higher amounts. Moreover, this substrate is transformed with high stereoselectivity to the corresponding (-)-anti-dihydrodiol epoxide by liver microsomes of Aroclor 1254-treated rodents. The metabolism results were in good accordance with the extent of stable adduct formation in calf thymus DNA as investigated by the 32P-postlabeling technique and with the mutagenicity in Chinese hamster V79 cells of the two enantiomeric 11,12-dihydrodiols mediated by hepatic postmitochondrial preparations of Aroclor 1254-treated rats. The results indicate that both genotoxic events occurred predominantly by the stereoselective activation of the (-)-(11R,12R)-dihydrodiol to the (-)-anti-DB[a,l]PDE with R,S,S,R-configuration. PMID- 9348440 TI - Adenine adducts with diepoxybutane: isolation and analysis in exposed calf thymus DNA. AB - 1,3-Butadiene (BD) is a high-volume industrial chemical and a common environmental pollutant. Although BD is classified as a "probable human carcinogen", only limited evidence is available for its tumorigenic effects in occupationally exposed populations. Animal studies show a surprisingly high sensitivity of mice to the carcinogenic effects of BD compared to rats (approximately 10(3)-fold), making interspecies extrapolations difficult. Identification and quantitation of specific BD-induced DNA adducts are important for improving our understanding of the mechanisms of BD biological effects and for explaining the observed species differences. Covalent binding of BD to DNA is probably due to its two epoxy metabolites: 3,4-epoxy-1-butene (EB) and 1,2:3,4 diepoxybutane (DEB). Both EB and DEB are direct mutagens producing frameshift and point mutations at both A:T and G:C base pairs. DEB is 100 times more mutagenic than EB and is found in quantity only in tissues of the most sensitive species (mouse). This has led to the suggestion that the higher sensitivity of mice to BD could be due to greater exposure to DEB. The present work was initiated in order to isolate and structurally characterize DEB-induced adenine adducts. The adducts were formed by reacting DEB with free adenine (Ade), 2'-deoxyadenosine (2'-dAdo), and calf thymus DNA followed by HPLC separation and analysis of the products by UV spectrophotometry, electrospray ionization mass spectrometry, and nuclear magnetic resonance. The adenine reaction resulted in three products which were identified as N-3-, N-7-, and N-9-(2'-hydroxy-3',4'-epoxybut-1'-yl)adenine. These adducts underwent acid-catalyzed hydrolysis to their corresponding (2',3',4' trihydroxybut-1'-yl)adenines upon heating or storage. The 2'-dAdo reaction with DEB followed by acid hydrolysis yielded a single adduct, N6-(2',3',4' trihydroxybut-1'-yl)adenine (N6-DEB-Ade). N-3-DEB-Ade and N6-DEB-Ade were also found in hydrolysates of calf thymus DNA exposed to DEB. The amounts of N-3-DEB Ade (13/10(3) normal Ade) and N6-DEB-Ade (5/10(3) normal Ade) were slightly lower than those of the corresponding EB-induced adducts in similar experiments, suggesting comparable reactivity of the two epoxy metabolites of BD toward adenine in DNA. The findings of this study provide a basis for future analyses of BD-induced adenyl DNA adducts in vitro and in vivo. PMID- 9348441 TI - Identification of adenine adducts formed in reaction of calf thymus DNA with mutagenic chlorohydroxyfuranones found in drinking water. AB - Calf thymus DNA was reacted with the extremely potent bacterial mutagen 3-chloro 4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) and the structurally related compounds 3,4-dichloro-5-hydroxy-2(5H)-furanone (MCA) and 3-chloro-4-methyl-5 hydroxy-2(5H)-furanone (MCF). The chromatograms of the HPLC analyses of the DNA hydrolysates showed peaks that represented adducted base moieties. It was possible to establish the structures of the adducts by comparing UV spectra and chromatographical properties of the DNA adducts with known adenosine and 2' deoxyadenosine adducts. The DNA adduct produced by MX was identified as 3-(2' deoxyribofuranosyl-N6-adenosinyl)propenal (M1A-dR). It was calculated that 1 nucleotide/10(5) nucleotides was converted to M1A-dR. The same adduct was formed also in the reaction of MX with 2'-deoxyadenosine (yield 0.01%). The M1A-dR adduct may play a role in the mutational events induced by MX in Salmonella typhimurium strain TP2428. The adducts produced in the reactions of MCA and MCF with DNA were identified as 3-(2'-deoxyribofuranosyl)-7-formylimidazo[2,1 i]purine (epsilon cA-dR) and 4-(2'-deoxyribofuranosyl-N6-adenosinyl)-3-formyl-3 butenoic acid (fbaA-dR), respectively. The yield of epsilon cA-dR was 5 adducts/ 10(6) nucleotides and of fbaA-dR 4 adducts/10(5) nucleotides. The biological significance of these adducts is unknown. PMID- 9348442 TI - Investigation of the adducts formed by reaction of butenedioic acids with adenosine. AB - Several genotoxic butenedioic acids present in chlorine-disinfected drinking water were allowed to react with adenosine, guanosine, and cytidine in aqueous solution. HPLC analyses, with detection at 254 and 310 nm, showed that clearly detectable products were formed only in the reactions with adenosine. The major products from the reactions between either 2-chloro-3-methyl-2-butenedioic acid (ox-MCF) or 2-chloro-3-(chloromethyl)-2-butenedioic acid (ox-CMCF) and adenosine were the same. This substance was isolated by C18 column chromatography and characterized by UV absorbance, 1H and 13C NMR spectroscopy, and mass spectrometry. It was identified as 3-(beta-D-ribofuranosyl)-7-carboxy-7-formyl-8 [9'-(beta-D-ribofuranosyl) -N6- adenosinyl]-1,N6-ethanoadenosine (cf epsilon A,A). The yields of cf epsilon A,A in reactions performed at pH 7.4 and 37 degrees C were 0.7% and 0.3% with ox-MCF and ox-CMCF, respectively. PMID- 9348443 TI - Identification of the major hepatic DNA adduct formed by the food mutagen 2-amino 9H-pyrido[2,3-b]indole (A alpha C). AB - 2-Amino-9H-pyrido[2,3-b]indole (A alpha C) is among the most prevalent heterocyclic amines detected in grilled or panfried meat; it was shown to be carcinogenic in mice, to induce preneoplastic foci in rat liver, and to form covalent DNA adducts in vitro and in vivo. The corresponding nitro compound 2 nitro-9H-pyrido[2,3-b]indole (N alpha C) was prepared and shown to be a direct acting mutagen in the Salmonella assay, while the amino compound required external metabolic activation with rat liver homogenate (S9). When A alpha C was incubated with S9 in the presence of calf thymus DNA, one major DNA adduct spot was detected upon 32P-postlabeling analysis. This adduct comigrated on ion exchange TLC and reversed-phase HPLC with the major adduct detected in primary hepatocytes treated with A alpha C. In DNA isolated from livers of male F344 rats treated with 800 and 160 ppm, the formation of the same major adduct was observed with relative adduct levels of 20.6 +/- 9.6 and 1.4 +/- 1.1 adducts/10(8), respectively, as determined with the butanol extraction variant of the 32P postlabeling assay. No DNA adducts were detected in liver DNA from rats treated with 32 ppm A alpha C or control animals. The major adduct spot was eluted and hydrolyzed and the modified base characterized by chromatographic and UV spectral comparison with a synthetic standard synthesized from acetylated guanine N3-oxide and A alpha C. Electrospray mass spectrometry and 1H- and 13C-NMR spectroscopy provided further evidence for the major adduct as N2-(guanin-8-yl)-2-amino-9H pyrido[2,3-b]indole. A alpha C is formed especially in high-temperature preparation of food and may contribute considerably to the human carcinogenic risk that might be imposed by heterocyclic amines. PMID- 9348444 TI - Dietary glycine prevents increases in hepatocyte proliferation caused by the peroxisome proliferator WY-14,643. AB - Peroxisome proliferators are a group of nongenotoxic carcinogens which include a number of hypolipidemic drugs, solvents, and industrial plasticizers. Although the mechanism by which they cause cancer remains unknown, one likely possibility is that they act as tumor promoters by increasing cell proliferation. Hepatic Kupffer cells represent a rich source of mitogenic cytokines (e.g., tumor necrosis factor alpha, TNF alpha) and are stimulated by peroxisome proliferators. Since glycine prevents activation of Kupffer cells, these experiments were designed to test the hypothesis that a diet containing glycine could block the mitogenic effect of the peroxisome proliferator [[4-chloro-6-(2,3 xylidino)pyrimidinyl]thio]acetic acid (WY-14,643). The effects of a glycine enriched diet on WY-14,643-induced increases in cell proliferation after a single dose or after feeding WY-14,643 in the diet for 3 weeks were assessed. As expected, 24 h after a single dose of WY-14,643, rates of cell proliferation increased from basal values of 0.7 +/- 0.3% to 5.1 +/- 0.5%. Glycine largely prevented the increase caused by WY-14,643 with proliferation only reaching 1.9 +/- 0.4% (p < 0.05). Acyl CoA oxidase increased from 1.4 +/- 0.1 to 3.5 +/- 0.6 nmol of H2O2 min-1 (mg of protein)-1 (p < 0.05) indicating that peroxisome specific enzyme activity was induced about 2-fold in livers of WY-14,643-treated rats after 24 h. Unlike cell proliferation, however, acyl CoA oxidase was not affected by dietary glycine, consistent with the hypothesis that cell proliferation and peroxisome proliferation occur via different mechanisms. After 3 weeks, dietary glycine reduced basal rates of cell proliferation by about 50% and completely prevented the sustained 5-fold increase in cell proliferation caused by feeding WY-14,643. Moreover, the 3-fold increase in TNF alpha mRNA caused by WY-14,643 was blocked completely by the glycine-enriched diet. Similarly, immunohistochemical staining for TNF alpha was increased 6-fold by WY 14,643, an increase which was prevented by dietary glycine. However, the 6-fold increase in acyl CoA oxidase activity was unaffected by glycine under similar conditions demonstrating that a diet containing 5% glycine prevents increased hepatocyte proliferation caused by a potent peroxisome proliferator without affecting induction of peroxisomes. These data demonstrate that a glycine enriched diet prevents stimulated cell proliferation most likely by inhibiting TNF alpha production and raise the possibility that dietary glycine will be effective in preventing cancer caused by nongenotoxic carcinogens such as WY 14,643. PMID- 9348445 TI - Both cytochromes P450 2E1 and 3A are involved in the O-hydroxylation of p nitrophenol, a catalytic activity known to be specific for P450 2E1. AB - 4-Nitrophenol 2-hydroxylation activity was previously shown to be mainly catalyzed by P450 2E1 in animal species and humans. As this chemical compound is widely used as an in vitro probe for P450 2E1, this study was carried out to test its catalytic specificity. First, experiments were carried out on liver microsomes and hepatocyte cultures of rat treated with different inducers. Liver microsomes from pyrazole- and dexamethasone-treated rats hydroxylated p nitrophenol with a metabolic rate increased by 2.5- and 2.7-fold vs control. Dexamethasone treatment increased the hepatic content of P450 3A but not that of P450 2E1. Two specific inhibitors of P450 3A catalytic activities, namely, ketoconazole and troleandomycin (TAO), inhibited up to 50% of 4-nitrophenol hydroxylation in dexamethasone-treated rats but not in controls. Hepatocyte cultures from dexamethasone-treated rats transformed p-nitrophenol into 4 nitrocatechol 7.8 times more than controls. This catalytic activity was inhibited by TAO. Similarly, hepatocyte cultures from pyrazole-treated rats hydroxylated p nitrophenol with a metabolic ratio increased by about 8-fold vs control. This reaction was inhibited by diethyl dithiocarbamate and dimethyl sulfoxide, both inhibitors of P450 2E1. Second, the capability of human P450s other than P450 2E1 to catalyze the formation of 4-nitrocatechol was examined in a panel of 13 human liver microsomes. Diethyl dithiocarbamate and ketoconazole reduced 4-nitrophenol hydroxylase activity by 77% (+/- 11) and 13% (+/- 16), respectively. Furthermore, the residual activity following diethyl dithiocarbamate inhibition was significantly correlated with seven P450 3A4 catalytic activities. Finally, the use of human cell lines genetically engineered for expression of human P450s demonstrated that P450 2E1 and 3A4 hydroxylated 4-nitrophenol with turnovers of 19.5 and 1.65 min-1, respectively. In conclusion, P450 3A may make a significant contribution to 4-nitrophenol hydroxylase activity in man and rat. PMID- 9348446 TI - Six month clinical evaluation of a biomimetic hydrogel contact lens. AB - PURPOSE: We performed a six-month, comparative, contralateral/bilateral study to assess the clinical performance of the Proclear contact lens, which is manufactured from a novel, biomimetic, 59% water content, hydrogel material (Omafilcon A). Omafilcon A is a hydrogel polymer and is based on a new biomimetic approach to the design of biomaterials for biomedical applications. The polymer incorporates a synthetic analogue of the phosphorylcholine (PC) headgroup, an important component found in all human cells. METHODS: Forty-two subjects were fit with the Proclear lens for 6 months of daily wear; 20 subjects wore a Proclear lens in one eye and a Permaflex lens in the fellow as a control, while 22 subjects wore Proclear lenses in both eyes. A one-step hydrogen peroxide system was used for disinfection and no protein remover was used. RESULTS: No subject was required to withdraw due to unacceptable physiological findings with the Proclear lens. At the 3-month visit, corneal staining was present in 33% of the Proclear eyes compared to 72% of the Permaflex-wearing eyes. At one month, the Proclear lens was graded significantly more comfortable than the Permaflex lens (P < 0.01), and all 11 of the subjects who expressed a preference did so in favor of the Proclear lens. Overall, the fitting characteristics and visual performance of the Proclear lens did not appear to vary during the study. Spectrofluorimetric analysis of worn lenses confirm that Proclear showed minimal protein and lipid spoliation, which was found to be independent of wear time and subject variations. In contrast, despite the use of a surfactant cleaner, Permaflex lenses showed measurable and significant levels of lipid spoilation. Despite significant differences in average mean thicknesses and the level of bulk water in the lenses, in vivo dehydration of the Proclear lens was significantly lower than that of the Permaflex lens. CONCLUSIONS: The results of this study have confirmed the viability of biomimetic PC-hydrogels in contact lens applications. In vivo and in vitro analyses tended to confirm that Omafilcon A has a number of superior material attributes, as compared to conventional hydrogels when used in contact lens applications. PMID- 9348447 TI - Cornea-contact lens interaction in the aquatic environment. AB - PURPOSE: A large population of ametropic scuba divers wear contact lenses. We discuss optics and corneal physiology, as well as the types of contact lenses that are appropriate for underwater activities. METHODS: We reviewed an extensive body of literature to formulate guidelines to aid the contact lens fitter in satisfying individual sport diver's needs. RESULTS: Optical factors such as image displacement and light wave-length shifts require that contact lenses for underwater use be suitably modified. Underwater images appear nearer and larger (requiring greater accommodation) and are made up almost exclusively of the short wavelength end of the spectrum. Correction of presbyopia, in particular, is influenced by these factors. For example, presbyopic contact lens-corrected myopes require greater near adds underwater than when viewing the same objects in air. In general, presbyopes should consider monovision correction to facilitate underwater visual tasks. Although divers wearing rigid gas permeable contact lenses run the risk of more corneal problems than soft lens wearers if conservative ascents are not adhered to, there are no compelling reasons to change lens types in patients who are already fully adapted. Soft contacts, while very stable on the eye during diving, present a greater risk of lens contamination by sea or fresh water exposure. However, the latter problems are easily overcome by using disposable soft lenses. CONCLUSION: In this paper, we present several suggestions for lens material, modifications required for underwater ametropia correction, and wearing modalities for the sport divers. An understanding of the dramatic changes that impact the properties of light, corneal physiology, and visual perception which accompany the diver below the surface will enable the contact lens fitter to design a lens appropriate to the needs of the individual patient. PMID- 9348448 TI - Hydrogel contact lens-corneal interactions: a new mechanism for deposit formation and corneal injury. AB - PURPOSE: In this study, we examined the interactions between hydrogel contact lenses and the cornea, and the role of these interactions in the pathogenesis of interfacial debris formation and the complications of contact lens use. METHODS: We used a corneal abrasion device to simulate the motion of contact lenses on the cornea and the ensuing abrasive interactions. We examined lens and corneal surfaces by Atomic Force Microscopy (AFM), Low Voltage Scanning Electron Microscopy (LVSEM), and optical microscopy (with vital staining of corneas) for unused hydrogel contact lenses, lenses tested in the corneal abrasion device, and worn contact lenses. Young's modulus of hydrogel contact lenses was also measured and compared with the modulus of the human cornea, as reported in the literature. RESULTS: We observed patterns of abrasive damage to the rabbit cornea in vitro caused by corneal interaction with hydrogel contact lenses. Comparison of AFM and SEM of unused lens surfaces with the surfaces of lenses tested in the abrasion device showed dramatic alterations of the contact lens surfaces. Damage to the lenses was also evident by AFM for lenses worn by volunteers. The modulus of hydrogel contact lenses was lower than the modulus of the human cornea. CONCLUSIONS: The surface morphology of hydrogel contact lenses is significantly altered during use. The Young's modulus of the cornea is higher than the modulus of hydrogel contact lenses. These observations suggest a new mechanism for contact lens complications; namely, damage to the contact lens by the cornea as an initial event that produces lens particles and deposits at the lens-cornea interface, followed by corneal abrasion and the onset of other complications. PMID- 9348449 TI - Clinical comparison of Omafilcon A with four control materials. AB - PURPOSE: The purpose of this study was to assess the clinical performance of hydrogel contact lenses manufactured from a novel, biomimetic, 59% water content, hydrogel material (Omafilcon A) as compared with a representative range of control materials (Polymacon, Etafilcon A, Atlafilcon A, and Welcon CE). METHOD: Thirty subjects were enrolled in a 2-month, double-masked, randomized, comparative, daily wear study. Subjects wore two sets of lenses; each pair was worn for 1 month. In each part of the study, subjects wore a Proclear (Biocompatibles Ltd.) test lens (Omafilcon A) in one eye and a control lens in the other eye. In vivo lens dehydration was measured after 1 week of lens wear. Lenses were collected at the end of the study and subjected to in vitro spectrofluorimetric analysis to measure protein and lipid deposits. RESULTS: Omafilcon A showed significantly less lipid spoilation than all the controls and significantly less protein than the Etafilcon A and Atlafilcon A lenses. High levels of protein were found on the Group 4 lenses (Etafilcon A) and high levels of lipid were found on the Group 2 lenses (Weicon CE). The Omafilcon A lenses exhibited significantly less on-eye dehydration than the Etafilcon A lenses (P < 0.0001) and the Weicon CE lenses (P < 0.005). The majority of subjects expressed a preference for the Proclear lens, generally for reasons of comfort. Some minor differences in aspects of lens fit were noted between the test and control lenses, however, none of these were felt to be great enough to affect comfort. CONCLUSIONS: We hypothesize that the reduced spoliation seen with the Omafilcon A lens relates to the biomimetic nature of the material which incorporates phosphorylcholine. PMID- 9348450 TI - Scleral lens induced corneal swelling: what is the effect of varying Dk and lens thickness? AB - PURPOSE: Scleral lenses remain an option for visual rehabilitation under some circumstances or when there are specific therapeutic indications. Wearing is often limited with polymethylmethacrylate lenses, but rigid gas permeable (RGP) scleral lenses offer a physiological improvement as measured with central corneal swelling. This study aimed to determine central corneal swelling with lenses of different oxygen transmissibilities and thicknesses. METHODS AND RESULTS: Central corneal swelling was measured in four normal subjects wearing sealed RGP scleral contact lenses for 3 hours. The lenses used were of Dk 32, 59, and 115 with thicknesses of 0.15, 0.30, 0.60, and 1.20 mm. A reduction in central corneal swelling was associated with an increase in Dk and a reduction in lens thickness (P < 0.05). This relationship was found to be non-linear for higher transmissibilities. CONCLUSIONS: The physiological improvement as measured by central corneal swelling was reduced with thin high Dk lenses, suggesting diminishing returns for further increases in Dk. For usual scleral lens thicknesses of 0.6 mm in a material with a Dk of 115, the mean central corneal swelling induced was less than 3%. PMID- 9348451 TI - Examination of contact lens surfaces by Atomic Force Microscope (AFM). AB - PURPOSE: We evaluated the use of Atomic Force Microscopy (AFM) in examining the surfaces of unused and worn hydrogel contact lenses under natural, fully hydrated conditions. METHODS: Using the AFM contact mode, we examined hydrogel lenses (Acuvue, Surevue, NewVues, CSI Clarity, SeeQuence) that were hydrated. RESULTS: Surface morphologies characteristics of each lens type and wear history were readily observed. The surfaces of worn lenses showed evidence of abrasion and altered morphology. These changes varied with type of contact lens and conditions of use and by site on the lens. CONCLUSIONS: AFM is a very powerful tool for high resolution examination of hydrated contact lens surface structure. The method avoids artifacts due to dehydration and coating which can occur even with low voltage Scanning Electron Microscopy. Significant differences in contact lens surface morphology were observed before and after wear. These observations may be of importance in helping develop improved new lens polymers and ocular solutions. PMID- 9348452 TI - Contact lens care using chlorhexidine acetate with ethyl-6-O-decanoyl-glucoside: a comparative clinical and bacteriological study. AB - PURPOSE: We compared Ethyl-6-O-decanoyl-glucoside 0.005% (EDG) combined with 0.00025% chlorhexidine acetate (EDGC) to a commercial polyaminpropylbiguanide (PAPB). METHODS: Fifty-nine subjects wearing both ionic and non-ionic contact lenses for 8-16 hours daily used either EDGC or PAPB as a cleaning and disinfectant agent. Neither mechanical nor separate cleaning agents were employed. The study period was for 8 weeks. The following symptoms were compared for each solution: blurred vision, dryness, foreign body sensation, redness, and dirty lenses. The following signs were also compared for each solution: conjunctival hyperemia, papillary hypertrophy, corneal deposits, purulence, limbal vascularization, subepithelial scarring, visual acuity, bulbar hyperemia, and tear breakup time. RESULTS: After 8 weeks, 52% of the subjects in the EDGC group showed no evidence of corneal or conjunctival abnormalities. In contrast, only 19% of the subjects in the PAPB group showed no abnormalities of the conjunctiva or cornea (P = 0.012). After 8 weeks, 25% of the EDGC group showed evidence of papillary hypertrophy, whereas 50% of the PAPB group showed similar findings (P = 0.007). In addition, after 8 weeks of wear, 21% of the subjects using EDGC had positive conjunctival cultures, whereas the rate of positive cultures in the PAPB group was 50% (P = 0.035). At the conclusion of the study, the protein contents of the lenses were 131 micrograms +/- 48 micrograms (N = 29) in the EDGC group and 185 micrograms +/- 65 micrograms (N = 26) in the PAPB group (P = 0.001). CONCLUSION: Subjects using EDGC had fewer pathological findings than subjects using PAPB as their cleaning and disinfecting agent. The mechanism by which EDGC reduced the rate of papillary hypertrophy needs further investigation. PMID- 9348453 TI - Risk factors and clinical outcomes between fungal and bacterial keratitis: a comparative study. AB - PURPOSE: Previous studies on fungal and bacterial keratitis were descriptive single case series analysis. We conducted a hospital-based retrospective study to evaluate fungal and bacterial keratitis using a case-control design to compare risk factors and clinical outcomes. METHODS: Twenty-nine cases of culture positive fungal keratitis seen over a 5-year period were compared to 51 cases of culture-positive bacterial keratitis seen over a 21 months period. Using bacterial keratitis as the reference group, case-control odds ratios (OR) for predisposing factors and cohort relative risks (RR) for clinical outcomes associated with fungal keratitis were derived. Mantel-Haenszel adjustment procedures were used to examine the respective roles of confounding and intermediate variables. RESULTS: Compared to bacterial keratitis, fungal keratitis was significantly more likely to be associated with ocular trauma (OR = 2.69, 95% confidence interval [CI], 1.06-6.86) but significantly less likely to be associated with contact lens wear (OR = 0.16, 95% CI, 0.04-0.67) and preexisting ocular diseases (OR = 0.23, 95% CI, 0.07-0.72). Fungal keratitis was more likely to perforate than bacterial keratitis (RR = 5.28, 95% CI, 1.35-20.66) and to require penetrating keratoplasty (OR = 5.86, 95% CI, 2.06-16.69). CONCLUSIONS: Fungal keratitis appears more likely to result from ocular trauma, whereas bacterial keratitis is more likely to result from contact lens wear and pre-existing ocular diseases. Fungal keratitis is more likely than bacterial keratitis to result in perforation and require penetrating keratoplasty. PMID- 9348454 TI - The effects of low pH and CO2 on bovine corneal lactate production. AB - PURPOSE: Hypoxic corneal swelling in human subjects is reduced by CO2 induced corneal acidosis. We investigated whether this effect could be explained by a reduction in lactate production under acidic conditions. METHODS: Lactate production from isolated bovine corneas was assayed under normoxic and hypoxic conditions at normal pH (7.5) and low pH (7.05). Low pH was achieved using a CO2/HCO3- Ringer or a HEPES buffered Ringer. RESULTS: Normoxic corneas produced significantly less (approximately 25%) lactate under all acidic conditions. In contrast, hypoxic corneas produced the same or slightly less lactate under acidic conditions (weighted average of all conditions = 6% less lactate). CONCLUSIONS: From these results, we concluded that pH can have a substantial effect on lactate production, but its influence on glycolytic activity during hypoxia is suppressed and may be negligible. PMID- 9348455 TI - Demographics and psychological implications for the aging population. AB - The aging of skin is more than a decline in physiologic functions. Aging skin is associated with regressive anatomical changes such as looseness, roughness, wrinkles, and dyspigmentations (age spots) which adversely affect appearance. Unattractive aged persons tend to have less confidence, have low self-esteem, and are in fact less healthy than those who have aged well. Physicians must educate the public to follow life-styles that prevent a prematurely aged appearance. Additionally, resources which make it possible to remarkably correct the various deteriorations of the photo damaged face are now available. Patients should be told that these interventions are not simply an expression of vanity, but add to self-esteem and improved social relations. PMID- 9348456 TI - Physiologic and structural changes associated with aging skin. AB - The spectrum of photoaging in human skin shows a recognizable progression from subtle to profound, reflecting the various anatomic and structural changes that appear with aging skin. Qualification and quantification of these changes on both the macroscopic and microscopic levels enable the physician to select and evaluate appropriate therapeutic approaches to the problem of chronic solar damage. Comparative studies remain to be done that will further refine our abilities to match the right solutions to the recognized problems. PMID- 9348457 TI - Topical treatment of the aging face. AB - This article discusses the various over-the-counter and prescription products available to help improve sun-damaged skin, as well as superficial peeling agents. Practical suggestions for treating patients are given. PMID- 9348458 TI - Cutaneous resurfacing. AB - Cutaneous resurfacing can be accomplished with application of acids, abrasive modalities, or the new generation of carbon dioxide lasers. Ultimately, the universal goal is removal and replacement of the epidermis and dermal collagen remodeling. The indications range from therapeutic and reconstruction to the treatment of the stigmata associated with senescence. The indications are not technique-specific, and the art of cutaneous resurfacing is identifying the cutaneous defect and selecting the appropriate tool or tools to realize the optimal clinical results. PMID- 9348459 TI - Preoperative assessment of the elderly patient. AB - Preoperative assessment of the elderly patient for surgery is vital to the success of the surgical procedure. A thorough evaluation must first begin with an understanding of the physiologic and pathophysiologic changes unique to the elderly patient and the aging skin. A complete preoperative assessment entails assessing the patient and dermatologic problem, preparing the patient and caregivers for the surgery and its expected outcomes, and highlighting issues and problems that need to be managed prior to the procedure. With the continued growth of the geriatric population, all dermatologic surgeons should be aware of the special issues related to their geriatric patients. With heightened awareness of and screening for potential pitfalls in the elderly surgical patient, adverse outcomes can be avoided. PMID- 9348460 TI - Treatment of facial furrows and rhytides. AB - Many substances have historically been used to address facial soft tissue defects. Currently in the United States autologous fat and injectable bovine collagen are the most commonly utilized injectable fillers. Additionally, the judicious application of BOTOX in the upper face as well as neck has all but revolutionized the use of filling agents in these locales. While other agents are briefly mentioned, this is an in-depth review of the characteristics and application of autologous fat, bovine collagen. PMID- 9348462 TI - Rejuvenation of the brow. AB - Aging of the upper face is a frequent complaint in patients seeking a more youth appearance. Sometimes muscle dynamics are excessively forceful and cause marked lines of expression relatively early in life. Regardless of chronologic age, it is imperative that the surgeon perform a problem-oriented, detailed analysis of the face and each subunit, so that imperfections can be individually assessed as they apply to the harmonic balance of the whole face. Various factors, such as degree of eyebrow ptosis, vertical dimension of the forehead, preexisting scars, and level of hairline, will determine the best approach to surgical correction of the aging brow. In the authors' opinion treatment of the forehead, when done together with a rhytidectomy, requires previous blocking of the cervicofacial flaps so as not to distort important anatomic landmarks when traction is applied to the frontal flap, respecting the patient's desire for a natural aesthetic result. PMID- 9348463 TI - Blepharoplasty and periorbital surgery. AB - Blepharoplasty is one of the most successful aesthetic surgical procedures. Careful preoperative planning and conservative tissue resections can help to minimize complications and optimize results. Although some young patients request blepharoplasty specifically because of age-related changes in the eyelid skin, the surgery is that of sculpture and contouring of the entire aesthetic unit. The aging process in the eyelid complex is characterized by skin texture changes with loss of elasticity and formation of wrinkles, fat redistribution, enophthalmos, and anterior displacement of fat with a lower eyelid orbital fat prolapse. Once the etiology of the deformity and the associated periorbital anatomy are recognized, a local assessment and surgical treatment plan can produce optimal results. PMID- 9348461 TI - Male pattern alopecia. Surgical options. AB - The surgical options for male pattern alopecia are discussed with emphasis on the most recent and widely accepted technique, combination micrografting. Candidate selection is standardized and emphasized, along with possible complications, techniques, and postoperative care. A variety of results are shown relative to candidate selection which demonstrate the natural and aesthetically pleasing nature of modern hair transplants. PMID- 9348464 TI - Facialplasty. AB - Facelift surgery is a highly satisfactory solution to many problems that are manifested in the aging face. With one operation, relatively inconspicuous incisions, and brief convalescence, correction of the mid-face and neck can be achieved, providing a uniform degree of facial rejuvenation. As long as our perception of youth and beauty continue to be admired, so will the demand for aesthetic surgery. PMID- 9348465 TI - The aging nose. AB - Mature adult rhinoplasty requires more sophisticated planning and flawless execution in order to achieve an optimal result. The improvement not only engenders a more pleasing feature, it also provides approximately 5 years' rejuvenation to the face. These patients possess thin skin, weak support structures, and vulnerable nasal function, rendering the rhinoplasty more enigmatic. The medical and emotional issues should be resolved prior to surgery. PMID- 9348466 TI - Rejuvenation of the perioral area. AB - Lip aging is characterized by a relaxation of the skin with creation of wrinkles and vermilion involution. Surgical techniques include skin excisions and vermilion reposition. PMID- 9348467 TI - Cosmetic options for the aging dentition. PMID- 9348468 TI - Surgical pearls in the management of the aging face from A to Z. AB - Most of the excellent plastic surgery textbooks are full of many new and important technical procedures that we all must familiarize ourselves with, evaluate, and consider for use in our own practices. But the differences in achieving an objectively good result versus a great result, or between a satisfied and a dissatisfied patient is often the result of employing a group of "little" concepts called "pearls". Learning the major maneuvers of a new technique, no matter how promising, will often fall short of achieving the ultimate goal, a satisfied patient. Some of the pearls are technical reminders, and others are more practice management oriented. PMID- 9348469 TI - Aesthetic considerations in patients of color. AB - This article focuses on the aesthetic cutaneous considerations in analyzing and treating the aging face in patients of color. Initially some of the physiologic differences between dark and light pigmented skin are elucidated. Hair conditions that lead to aesthetic unacceptance are discussed as are facial enhancing cosmetic procedures, with special emphasis on people of color. Information on treatment of dandruff, xanthelasma, and melasma in people with dark skin is also presented. PMID- 9348470 TI - Nonsteroidal treatment of autoimmune skin diseases. AB - This brief survey has, it is hoped, helped to ease some of the anxiety associated with the management of complex autoimmune skin disorders. The main points to remember are that there are numerous therapeutic options for each disease. I like to think of this method of therapeutics as 'informed trial and error.' One does not need to master the monitoring, side-effect profile, or dosing regimen for each of the drugs in Table 1; only a working knowledge of a few is sufficient. I hope that the readers take advantage of the many tables and caveats I have included so this article can be a ready reference for many future uses. PMID- 9348472 TI - The application of the Gompertz model to describe body growth. AB - Three potential errors in applying the Gompertz growth model are discussed: 1. Restriction of growth data to the subadult or juvenile phase; 2. Confusion of mean growth rate with (mean) maximum growth rate; 3. Improper application of the growth model in cases of sample size variation. These errors are demonstrated on the example of growth data of Zambian common mole-rats (Cryptomys sp.). PMID- 9348471 TI - Paralysis and long bone growth in the chick: growth shape trajectories of the pelvic limb. AB - Growth of chick embryonic femora, tibiotarsi and first phalanges of digit three were measured at one day intervals from day 6 through 16 of incubation. Normal controls were compared to embryos paralyzed at 5 days of incubation. Over the 10 day study period, length of the paralyzed femora, length and width of the paralyzed tibiotarsi and differences in length of the phalanges were observed. Growth in length of phalanx one of digit three was most affected by paralysis over this period. Changes in shape of these bones also occurred during growth. Normal long bones undergo changes in shape as differential growth in length and width occurs. Such changes in shape can be considered as the bone's normal growth "trajectory". Paralyzed bones displayed a different growth trajectory than normal bones. As expected, the long bones of paralyzed embryos were shorter than age matched controls. Contrary to expectations, however, paralyzed long bones were relatively more stout than age-matched controls. PMID- 9348473 TI - Energy balance through age-related diurnal body weight variations in genetic obese and nonobese rats. AB - Indirect evidence of energy balance in laboratory rats is provided through the study of diurnal body weight variations in two inbred lines: obese beta and nonobese alpha, from birth to 200-300 days of age, with different feeding patterns from 25 to 75 days of age. Nocturnal weight gain (NWG) was the gain recorded after the dark phase, in direct relation to the acquisition of exogenous calories in excess of the current metabolic expenditure at nighttime. Daytime weight variation was either weight gain during lactation (DWG) or weight loss from weaning onwards (DWL), recorded after the light phase. DWL is in direct relation to daytime energy output, when metabolic expenditure exceeds the low rate of acquisition of exogenous calories. The correlation between averaged individual DWL and NWG absolute values was highly significant at every age studied. An increase in absolute DWL values with age was observed and at adulthood DWL was compensated for equivalent NWG. This increasing energy output with age during daytime, is most likely related to the maintenance of increasing biomass and consequently, to progressive reduced growth energy availability. The existence of energy homeostasis and ponderostat with set points genetically prescribed in adults, is suggested. Significant differences between lines found before adulthood give indirect evidence of higher fat accretion in the obese line in those periods of intense growth, known as the active phase of obesity. PMID- 9348474 TI - Influence of sex steroids on development of cultured fetal rat metatarsal bones. AB - The effects of 17 beta-estradiol (E), dihydrotestosterone (D, a non aromatisable androgen), and progesterone (P) on osteogenesis were studied on fetal rat cartilaginous anlagues cultivated in vitro. The three medial metatarsal rudiments were harvested at day 19 of gestation and grown in 1% BSA MEM medium (MO 643, Sigma) without serum nor antibiotics. After a 18h preincubation period, hormones were added for 8 days. Paired controls were incubated in the same volume of medium. The length, the metacarpal thickness and the size of the mineralized zone were measured every day, using a calibrated eyepiece (magnification X 40). DNA and protein synthesis, cartilage metabolism and mineralization were evaluated by monitoring the incorporation of 3H-Thymidine, 3H-Proline, 35S and 45Ca into anlagues for the last three hours of incubation, respectively. The dose/response effect of each steroid was studied at the concentrations of 10(-4) M, 10(-6) M, 10(-7) M and 10(-9) M. No difference was observed between male and female fetuses. A significant positive effect on total length (% of length measured at harvesting day) was observed with the 10(-7) M dose of E (163% +/- 2 vs 148% +/- 4 in controls) or D (158% +/- 3). Endochondral growth was not modified by P treatment. The effect of the three steroids (given at a dose of 10(-7) M) alone or as combinations (E, D, P, EP, ED, PD, EPD) confirmed the positive effect of E on endochondral growth and, to a lesser extend, of D and the association ED. Nevertheless, D had a better effect than E on endomembranous growth. On the contrary, P did not affect growth neither administrated alone nor in combination with E or D, while a positive effect of P on mineralization was demonstrated. The treatment associating the three steroids slowed down all the parameters concerning growth but strongly stimulated calcification. PMID- 9348475 TI - Different racemization ratios in dentin from different locations within a tooth. AB - It has been reported that the amount of D-aspartic acid in dentin is highly correlated with age. However, further studies are necessary to determine the details of this correlation. We quantitatively determined L- and D-aspartic acids in dentin, and from the data calculated the D/L ratio. We did not find any statistically significant difference in the D/L ratio between the same type of left and right teeth from the same jaw, and between the vestibular and lingual sides of the root dentin. However, the D/L ratio was significantly higher on the lingual side than on the labial side of the dentin. Although the D/L ratio in young subjects was comparatively high in the tooth crown and decreased toward the apex of the root, we did not always observe such a tendency in middle- to advanced-age individuals. These results indicate that the ratio of D/L aspartic acid in dentin varies between the lingual side and vestibular side of the crown dentin. Therefore, it appears that the racemization rate of aspartic acid is not uniform but differs with the region of the dentin, and may be affected by differences in factors such as temperature. PMID- 9348476 TI - Computerised control of growth and measurement of metabolic rate. PMID- 9348477 TI - Predicting student satisfaction in baccalaureate nursing programs: testing a causal model. AB - The purpose of this study was to test a causal model for predicting the overall satisfaction of senior students with their baccalaureate nursing programs. The proposed model was primarily a sociological impact model based upon Tinto's (1975) student integration theory for predicting student departure, and Pascarella's (1985) causal model for predicting student outcomes. To test the model, a sample of 195 senior female students from five baccalaureate nursing programs in the southwestern United States was selected for step-wise regression studies and path analysis. Testing the causal model, the students' integration into the academic and social systems of their nursing programs directly explained 42% of the variance for predicting overall satisfaction. The best predictors of overall satisfaction were the students' 1) academic development, 2) satisfaction with facilities and services, 3) satisfaction with the faculty, and 4) social interaction with peers. PMID- 9348478 TI - A comparison of predictors of success on NCLEX-RN for African American, foreign born, and white baccalaureate graduates. AB - This retrospective study was concerned with identifying the strongest predictors of success for African American and foreign-born baccalaureate graduates on the NCLEX-RN from the following nine variables--the admission grade point average, medical-surgical nursing grade point average, nursing grade point average, cumulative grade point average, percentile rank on the Mosby Assess Test, age at the time of the licensing examination, number of semesters needed to complete the nursing curriculum, licensed vocational nurse status, and the number of Ds and Fs received in nursing courses--and comparing these with predictors of success for white baccalaureate graduates. Three random samples of 50 African American, foreign-born, and white graduates from four baccalaureate schools from May 1987 through May 1992 were selected from a total population of 1,205. Chi-square, Fisher's exact test, two-way analysis of variance, and discriminant analysis were used to analyze the data. Students in all three ethnic groups with a Mosby Assess Test percentile rank below 21 and a D or F in a nursing course were more likely to fail the NCLEX-RN than those with a higher percentile rank and no Ds or Fs. PMID- 9348479 TI - Identification, accommodation, and success of students with learning disabilities in nursing education programs. AB - Few studies have addressed the issue of nursing students with learning disabilities, although students with both identified and undiagnosed learning disabilities are pursuing nursing education. Legal mandates concerning these students impact nursing programs and faculty. To reduce the risk of discrimination litigation, nursing education programs need to establish educational strategies to promote these students' success. The purpose of this research was to discover the extent to which Bachelor of Science in Nursing and Associate Degree of Nursing programs in one southeastern state admit, identify, and graduate nursing students with learning disabilities, and to identify accommodations provided by these programs to promote success among this student population. Of the 54 programs surveyed, 45 responded. Almost 50% indicated that their program had admitted nursing students with learning disabilities and one third reported graduating students with learning disabilities. Enrolled students with undiagnosed learning disabilities were identified during their course of studies by both faculty members and by students themselves. The most frequently reported accommodations for students were counselors, tutors, tape-recorded lectures, and computer access. As the number of students with learning disabilities seeking post-secondary education increases, nursing programs and nurse educators will be involved with greater numbers of students needing educational accommodations. PMID- 9348480 TI - NCLEX-RN: from job analysis study to examination. AB - The National Council Licensure Examination for Registered Nurses (NCLEX-RN) is used by boards of nursing in licensure decision making and must be job related and reflect current entry-level nursing practice. This article describes the rigorous process used to develop the licensure examination for registered nurses. PMID- 9348481 TI - Students' perceptions of nursing: their relationship to attrition. AB - This article describes a cross-sectional study that examined the effect of students' pre-enrollment perceptions of nursing education on attrition. It was hypothesized that more students who leave (leavers) than students who continue would report a discrepancy between these perceptions and their nursing academic experience. It was also hypothesized that leavers would rate several potential stressors as more important concerning decisions to leave than would continuers. The sample comprised students who commenced their nursing education at an Australian tertiary institution. As predicted, a greater percentage of leavers than continuers reported nursing content to differ from what they expected. The major area of conflict was the scientific component in nursing knowledge. The groups did not differ concerning potential stressors' influence on decisions to leave. While constrained by its cross-sectional nature, this study's findings suggest a need to adequately convey the scientific basis of nursing knowledge to potential students and to deal with misconceptions early in education, to reduce attrition. PMID- 9348482 TI - Returning to school to a baccalaureate program: is there an easy way to learn? AB - Registered nurses returning to school need to learn many new skills in their first courses at the university. The assignment presented here assisted students in learning the course content and honing other skills needed for their academic life. Design and implementation of the assignment as well as comments and recommendations for change will be given. PMID- 9348483 TI - Young people considering nursing as a career: starters vs. non-starters. AB - This study focused on applicants who were accepted to a nursing education program but did not begin their studies, thus wasting time, effort and money. Of 953 accepted applicants, 27% did not enter the program. Significant variables differentiating starters from non-starters were higher psychometric score, higher paternal education, lower priority for nursing studies, older age, non-Israeli country of birth and not completed army service. The study relating to starters vs. non-starters is one aspect of a large study (Ehrenfeld, Rotenberg, Sharon, & Bergman, 1995). In most nursing education programs the number of applicants exceed the faculty's capacity, and suitable candidates must be rejected. It is therefore both wasteful and disappointing when accepted applicants change their mind and do not actually begin in the program. In light of the costs and complexities involved in the student screening process and the continuous efforts extended today all over the world to raise the standards of nursing education and care, the factors differentiating starters from non-starters may have important implications. The aim of the present study was to evaluate the screening process at the Tel Aviv University baccalaureate nursing program and determine which variables were predictive of starters and non-starters among accepted applicants. PMID- 9348484 TI - Student evaluation of courses: determining the reliability and validity of three survey instruments. AB - The purpose of this study was to determine the reliability and validity of three instruments designed for student evaluation of courses: the Course, Library, and Computer (CLC); the Clinical (CLIN); and the Nursing Skills Lab (NSL) evaluation tools. Using data from 294 anonymous associate degree nursing students, internal consistency and reliability were demonstrated by the high alpha coefficients ranging from .85 to .94. An exploratory principal component analysis, with the varimax rotation, provided an estimate of construct validity. PMID- 9348486 TI - Personality disorder assessment: the challenge of construct validity. AB - We begin with a review of the data that challenge the current categorical system for classifying personality disorder, focusing on the central assessment issues of convergent and discriminant validity. These data indicate that while there is room for improvement in assessment, even greater change is needed in conceptualization than in instrumentation. Accordingly, we then refocus the categorical-dimensional debate in assessment terms, and place it in the broader context of such issues as the hierarchical structure of personality, overlap and distinctions between normal and abnormal personality, sources of information in personality disorder assessment, and overlap and discrimination of trait and state assessment. We conclude that more complex conceptual models that can incorporate both biological and environmental influences on the development of adaptive and maladaptive personality are needed. PMID- 9348485 TI - Introduction to the special feature: research directions for the personality disorders: Part I. PMID- 9348487 TI - A behavior analytic conceptualization of personality disorders: a response to Clark, Livesley, and Morey. AB - Clark, Livesley, and Morey critically analyze some of the deficiencies in the DSM approach to classifying personality disorders that derive from the fact that the construct of personality disorders is inadequately specified. This response agrees with their criticism, but argues that any significant improvement will have to entail a theory based taxonomy. A taxonomic system must eventually be explained rather than be an explanation for behavior. The theory chosen must relate to the goals of those creating the classification system. This paper suggests that a taxonomic system that is tied to an experimentally based set of behavioral principles, and can provide treatment utility, would have advantages over DSM-IV. The paper than presents the outline of a classification system founded on behavior analytic principles. PMID- 9348488 TI - Extent of comorbidity between mental state and personality disorders. AB - Comorbidity between major psychiatric disorders (Axis I) and personality disorders (Axis II) is widespread, often extremely strong, and invariably confusing. The strongest associations are found between substance use and the cluster B (flamboyant) personality disorders, anxiety disorders and the anxious/fearful personality group (cluster C), and between somatisation and both cluster B and C disorders. The significance of these associations is far from clear, and almost certainly include more than one type of relationship. Empirical studies of patients with and without Axis I and II comorbidity show that the presence of a personality disorder can affect the outcome of treatment, both positively and negatively, in a way that currently appears unpredictable. One useful way of interpreting this comorbidity is by postulating personality dispositions that make some people prone to certain mental state disorders. PMID- 9348489 TI - Heuristic models of comorbidity of axis I and axis II disorders. AB - The mechanisms responsible for the co-occurrence of personality disorders with Axis I disorders are not well understood. We propose a number of models that include various relationships at the etiological, pathophysiological, and observable clinical levels between personality disorders and Axis I disorders. It is recommended that such models be subjected to empirical tests to assess their validity. PMID- 9348490 TI - Psychometric characteristics of the cluster B personality disorders under DSM-III R and DSM-IV. AB - A stated goal for the latest revision of the DSM was improving the performance of the Axis II system. To evaluate the degree to which this goal was achieved, we performed a psychometric analysis of the Cluster B personality disorders (PD) as they are defined under the DSM-III-R and its successor the DSM-IV. Ninety-four patients with a primary PD diagnosis were rated for the presence of DSM-III-R and DSM-IV Cluster B PD criteria. From this symptom level data, the convergence, divergence, and internal consistency of the Cluster B criteria sets were determined. Also, kappa values were computed between the DSM-III-R and DSM-IV versions of these disorders as an index of coverage or agreement across the two systems. The results indicated that, in general, the DSM-IV Cluster B PDs represent an improvement over their DSM-III-R predecessors. However, some psychometric limitations, particularly regarding the convergence of the criteria sets, continue to be present. PMID- 9348491 TI - Interrater reliability and internal consistency of the structured clinical interview for DSM-IV axis II personality disorders (SCID-II), version 2.0. AB - Interrater reliability and internal consistency of the SCID-II 2.0 was assessed in a sample of 231 consecutively admitted in- and outpatients using a pairwise interview design, with randomized rater pairing and blind interview assessment. Interrater reliability coefficients ranged from .48 to .98 for categorical diagnosis (Cohen kappa), and from .90 to .98 for dimensional judgements (Intraclass correlation coefficient). Internal consistency coefficients were satisfactory (.71-.94). The results suggest that the SCID-II 2.0 has adequate interrater and internal consistency reliability. PMID- 9348492 TI - Negative and positive dimensions of schizotypal personality disorder. AB - The positive (perceptual-cognitive) and negative (social-interpersonal) dimensions of schizotypal personality traits were examined in biological relatives of individuals with Axis I disorder. The subjects were young adult offspring from three contrasting parental groups, including schizophrenic disorder, affective disorder, and normal controls. Cognitive correlates, including digit span (presumed to assess working memory) and P3 amplitudes, were also examined. Preliminary results showed that positive and negative dimensions were distinguished by different prevalence patterns in the offspring subjects, and by a different pattern of correlations with cognitive measures. Negative dimensions were more frequent in offspring from the schizophrenic parental group than in the offspring from affective disorder and normal control parental groups. Digits forward and backward, and P3 amplitude decrements, characterized a subset of offspring with negative features from the schizophrenic parental group. Positive dimensions did not differ between the psychiatric parental groups, and did not covary with digit span or P3 amplitude assessments. These results support the view that positive and negative dimensions may reflect separable pathophysiologic processes. PMID- 9348493 TI - Pheochromocytoma in dogs: 61 cases (1984-1995). AB - This report presents the clinical, laboratory, imaging, and pathologic findings in 61 dogs with pheochromocytoma by retrospective evaluation of medical records. Pheochromocytomas were diagnosed by histopathologic examination of tissue specimens in all dogs. Special stains (chromogranin A and synaptophysin) also were used to confirm the chromaffin cell origin of the tumors. Epidemiologic findings were in agreement with previous studies, indicating that pheochromocytomas affect middle-aged to older dogs with no apparent gender or breed predilection. The tumor was considered clinical in 21 dogs (34%), was responsible for abnormalities related to a space-occupying mass in 7 dogs (11%), and was an incidental finding in 35 dogs (57%). The hematologic and biochemical findings were nonspecific. Hypertension was detected in 10 of 23 (43%) dogs tested, but all hypertensive dogs had concurrent diseases that may have contributed to hypertension. Abdominal ultrasonography was the most commonly used imaging procedure, with a mass detected in the region of the adrenal glands in 20 of 40 (50%) dogs examined. In 4 of the 20 dogs (20%), invasion of the caudal vena cava was identified. Surgery was performed in 17 dogs (28%) with immediate death or euthanasia of 5 dogs. Survival after surgery ranged from 1 day to 3.25 years. Pheochromocytomas were locally invasive in 39% of affected dogs and produced metastases in 13% of the cases. Common sites for metastases included regional lymph nodes, liver, lung, kidney, spleen, and bone. A high frequency of concurrent neoplasia (54%), including endocrine neoplasia, was identified. PMID- 9348495 TI - Fluoroscopically guided percutaneous disk aspiration in 10 dogs with diskospondylitis. AB - Fluoroscopically guided percutaneous fine-needle aspiration of the intervertebral disk space was performed in 10 dogs with diskospondylitis. Positive bacterial cultures were obtained from 9 of 12 aspirated disk spaces, 1 of 6 blood cultures, and 6 of 10 urine cultures. Positive disk cultures were obtained from 2 dogs with negative blood and urine cultures and from 2 additional dogs with low numbers of Staphylococcus in urine cultures. Adverse clinical sequelae of the procedure were not noted. Percutaneous fine-needle aspiration of the intervertebral disk space is an alternative technique to surgical biopsy to obtain positive bacterial cultures from dogs with diskospondylitis. PMID- 9348494 TI - Phase II clinical trial of carboplatin in canine transitional cell carcinoma of the urinary bladder. AB - Fourteen dogs with histologically-confirmed transitional cell carcinoma (TCC) of the urinary bladder were treated with 300 mg/m2 carboplatin every 3 weeks. Response to therapy was assessed with abdominal radiography, double contrast cystography, urinary bladder ultrasonography and thoracic radiography before therapy and at 6-week intervals during therapy. Dogs were monitored for hematologic toxicity with a CBC and platelet count performed immediately before and 10 to 14 days after carboplatin treatment. Tumor responses included progressive disease in 11 dogs and stable disease in 1 dog. Two dogs were euthanized due to carboplatin toxicity before assessment of tumor response. Toxicity included thrombocytopenia with or without neutropenia in 7 dogs and gastrointestinal toxicity in 6 dogs. Carboplatin therapy was not beneficial in the treatment of TCC in the 14 dogs in this study. PMID- 9348496 TI - Silica-containing urinary calculi in dogs (1981-1993). AB - Silica-containing urinary calculi obtained from 773 dogs and submitted by veterinarians throughout the United States were analyzed by quantitative crystallographic analysis to determine mineral composition. Specimens were composed of either multiple mineral layers (535 specimens) or 1 mineral layer (238 specimens). Most multiple-layer calculi were composed of 80% or greater silica (300 of 535, 56%) or 20% to 79% silica (184 of 535, 34%) in any mineral layer. Most 1-layer calculi were composed of 100% silica (212 of 238, 89%). Most dogs forming silica-containing calculi were of male gender (679 of 773, 88%). Bacterial cultures of calculus or urine or both were performed on 49% (376 of 773) of the specimens, and bacterial growth was obtained from 37% (139 of 376) of samples cultured. The prevalence of calculus-associated urinary tract infection was 35% (113 of 321) in males and 47% (26 of 55) in females. The gender prevalence for infection with Staphylococcus species was 16% (51 of 321) in males and 33% (18 of 55) in females. The breed and gender of dogs that formed calculi (silica population) were compared with the hospital population (Veterinary Medical Teaching Hospital [VMTH] population) and with a population of calculus forming dogs (Stonelab population) to determine risk factors for silica calculus formation. For all breeds compared, the ratio of males to females was higher in the silica population. The German Shepherd Dog and Old English Sheepdog were significantly overrepresented when the silica population was compared with either the VMTH population or the Stonelab population. We conclude that male German Shepherd Dogs and Old English Sheepdogs are at increased risk for formation of silica-containing urinary calculi. PMID- 9348498 TI - Polyuria and polydipsia and disturbed vasopressin release in 2 dogs with secondary polycythemia. AB - In dogs, secondary polycythemia (SP) may be associated with polyuria and polydipsia (PU/PD). The pathogenesis of this PU/PD has not yet been explained. We hypothesized that hyperviscosity and increased blood volume in SP might affect vasopressin (VP) release, resulting in PU/PD. This hypothesis was tested in 2 dogs with SP caused by renal neoplasia and PU/PD. Osmoregulation of VP release was studied by a modified water deprivation test and by investigating the VP response to hypertonic saline infusion. Water deprivation test results were consistent with an inability to produce concentrated urine despite increasing plasma osmolality. During hypertonic saline infusion, the osmotic threshold of VP release was markedly increased in both dogs, resulting in a delayed VP response to increasing plasma osmolality. The sensitivity of VP release was low normal in both dogs. We conclude that blood hyperviscosity and increased blood volume led to impaired VP release and polyuria. PMID- 9348497 TI - Clinical and clinicopathologic effects of large doses of raw linseed oil as compared to mineral oil in healthy horses. AB - The clinical and clinicopathologic effects of raw linseed oil and mineral oil were compared. In a crossover experimental design trial, 6 horses were given either raw linseed oil (2.5 mL/kg body weight) or mineral oil (10 mL/kg body weight), twice, 12 hours apart. Two weeks later, the horses received the opposite treatment. All horses given mineral oil or linseed oil developed nonformed feces by 24 hours of the first administration of oil. Horses treated with mineral oil had formed feces at 48 hours; horses treated with linseed oil developed normally formed feces at 96 to 108 hours. All horses treated with linseed oil had signs of depression and anorexia, and 3 had signs of mild colic. These signs were not observed in horses treated with mineral oil. Concentrations of serum glucose and bilirubin were significantly higher in horses treated with linseed oil when compared with horses treated with mineral oil. PMID- 9348499 TI - Neonatal alloimmune thrombocytopenia in a quarter horse foal. AB - Neonatal alloimmune thrombocytopenia is recognized as a spontaneous disease of human infants, piglets, and possibly mules, but it has not been previously reported in horses. A 1-day-old Quarter Horse foal presented to Michigan State University Large Animal Clinic with severe thrombocytopenia of unknown origin. Immunoglobulins that bound to the foal's platelets were identified in the mare's plasma, serum, and milk by indirect assays. The immunoglobulins were further shown to recognize platelets from the foal's full brother, born 1 year earlier. These findings, coupled with the clinical course of the foal during its period of hospitalization, strongly suggest that neonatal alloimmune thrombocytopenia can spontaneously occur in neonatal horses. This diagnosis should be considered for foals with severe thrombocytopenia when other causes can be excluded, and platelet antibody assays should be used to support this diagnosis. PMID- 9348500 TI - Thrombocytopathia and light-chain proteinuria in a dog naturally infected with Ehrlichia canis. AB - A 6-year-old dog was presented for evaluation of recurrent epistaxis. Platelet counts, biochemical tests, and coagulation tests were within the normal range, but a mucosal bleeding time was prolonged; there was hyperproteinemia and a monoclonal gammopathy. Heterogeneity of light chains appeared in urine, however, thus suggesting that the gammopathy was polyclonal. Platelet aggregation tests showed decreased responsiveness to collagen. An Ehrlichia canis indirect fluorescent-antibody titer was positive (1:40). Treatment with tetracycline, melphalan, and prednisone resulted in a rapid clinical improvement that persisted for at least 3 years. PMID- 9348501 TI - Isolation and characterization of karyoskeletal protein-enriched fractions from vertebrate livers. PMID- 9348502 TI - Preparation of karyoskeletal protein-enriched fractions from Drosophila melanogaster cells and tissues. PMID- 9348504 TI - Determining nuclear structure using the fluorescence light microscope. PMID- 9348505 TI - Localization of single nuclear pore complexes by confocal laser scanning microscopy and analysis of their distribution. PMID- 9348503 TI - Isolation of nuclei and nucleoli from the yeast Saccharomyces cerevisiae. PMID- 9348506 TI - Nuclear ultrastructure: transmission electron microscopy and image analysis. PMID- 9348507 TI - Three-dimensional surface structure analysis of the nucleus. PMID- 9348508 TI - Scanning transmission electron microscopy of nuclear structures. PMID- 9348509 TI - Electron microscopic imaging of chromatin with nucleosome resolution. PMID- 9348510 TI - Mapping three-dimensional chromosome architecture in situ. PMID- 9348511 TI - Structural analysis of meiotic chromosomes and synaptonemal complexes in higher vertebrates. PMID- 9348512 TI - Genetic and morphological approaches for the analysis of meiotic chromosomes in yeast. PMID- 9348513 TI - Mapping proteins to nuclear pore complexes by immunogold electron microscopy. PMID- 9348514 TI - Methods used to study structure and function of the nucleolus. PMID- 9348515 TI - Identification of base-unpairing region-binding proteins and characterization of their in vivo binding sequences. PMID- 9348517 TI - Analysis of nuclear envelope assembly using extracts of Xenopus eggs. PMID- 9348516 TI - Cell-free nuclear reassembly in mammalian mitotic homogenates. PMID- 9348518 TI - Cell-free nuclear assembly and disassembly in Drosophila. PMID- 9348520 TI - Mapping of DNA replication sites in situ by fluorescence microscopy. PMID- 9348519 TI - Methods for studying in vitro assembly of male pronuclei using oocyte extracts from marine invertebrates: sea urchins and surf clams. PMID- 9348521 TI - EM visualization of transcriptionally active genes after injection into Xenopus oocyte nuclei. AB - This article has described methods in use in our lab for microinjection of genes into Xenopus oocyte nuclei followed by EM visualization of those genes by the Miller chromatin spreading method. We consider our efforts to be still developing, as we attempt to maximize the visualization of specific, active, mappable genes. One of our main goals at this time is to find a DNA sequence element that will ensure efficient Pol II termination so that the common problem of read-through transcription (as seen in Fig. 6) can be overcome. We currently are testing three different elements reported to have roles in transcription termination. The method is evolving as a unique and valuable approach to study gene expression and RNA processing at the level of individual genes and individual transcripts. Given the ability to manipulate both cis- and trans acting factors prior to EM visualization, its potential is limited only by the somewhat labor-intensive nature of the method. PMID- 9348522 TI - Cell-free systems to study chromatin remodeling. PMID- 9348523 TI - In vitro systems for the reconstitution of snRNP and protein nuclear import. AB - In this chapter we have presented the most recent methods for the preparation of cell extracts and recombinant protein factors for the reconstitution of nuclear protein and snRNP import in vitro. In addition, we have discussed methods available for the quantitation of the level of import into nuclei. Accurate quantitation is particularly important when the effects of inhibitors are to be compared and when estimates of nuclear import rate are required. PMID- 9348524 TI - In vivo nuclear transport kinetics in Saccharomyces cerevisiae. AB - We have described a direct fluorescence assay to measure the relative rates of NLS-directed import and passive export of an NLS-GFP fusion protein in yeast. The design and construction of the reporter GFP fusion, its spectral qualities, size, use of inducible promoters, and the choice of NLS, are variables that could extend the method's utility. Future applications will almost certainly demand the quantification of transport rates in single cells using image analysis techniques. As is the case whenever cellular processes are studied in vivo, the in vivo nuclear trafficking properties of NLS-GFP are complicated and poorly understood. Some will be attracted to NLS-GFP kinetic assays simply because so little is known about the function and regulation of the transport apparatus in living cells. At the same time, the uncertainties that accompany in vivo work necessarily prevent the rigorous interpretation of data, which biochemists expect from experiments performed in vitro using highly purified enzymes. PMID- 9348525 TI - Nuclear transport of RNAs in microinjected Xenopus oocytes. PMID- 9348526 TI - In vivo systems to study the dynamics of nuclear lamins. PMID- 9348527 TI - The Rho-GEF Rom2p localizes to sites of polarized cell growth and participates in cytoskeletal functions in Saccharomyces cerevisiae. AB - Rom2p is a GDP/GTP exchange factor for Rho1p and Rho2p GTPases; Rho proteins have been implicated in control of actin cytoskeletal rearrangements. ROM2 and RHO2 were identified in a screen for high-copy number suppressors of cik1 delta, a mutant defective in microtubule-based processes in Saccharomyces cerevisiae. A Rom2p::3XHA fusion protein localizes to sites of polarized cell growth, including incipient bud sites, tips of small buds, and tips of mating projections. Disruption of ROM2 results in temperature-sensitive growth defects at 11 degrees C and 37 degrees C. rom2 delta cells exhibit morphological defects. At permissive temperatures, rom2 delta cells often form elongated buds and fail to form normal mating projections after exposure to pheromone; at the restrictive temperature, small budded cells accumulate. High-copy number plasmids containing either ROM2 or RHO2 suppress the temperature-sensitive growth defects of cik1 delta and kar3 delta strains. KAR3 encodes a kinesin-related protein that interacts with Cik1p. Furthermore, rom2 delta strains exhibit increased sensitivity to the microtubule depolymerizing drug benomyl. These results suggest a role for Rom2p in both polarized morphogenesis and functions of the microtubule cytoskeleton. PMID- 9348528 TI - Endoplasmic reticulum stress-induced mRNA splicing permits synthesis of transcription factor Hac1p/Ern4p that activates the unfolded protein response. AB - An intracellular signaling from the endoplasmic reticulum (ER) to the nucleus, called the unfolded protein response (UPR), is activated when unfolded proteins are accumulated in the ER under a variety of stress conditions ("ER stress"). We and others recently identified Hac1p/Ern4p as a transcription factor responsible for the UPR in Saccharomyces cerevisiae. It was further reported that Hac1p (238 aa) is detected only in ER-stressed cells, and its expression is mediated by unconventional splicing of HAC1 precursor mRNA. The splicing replaces the C terminal portion of Hac1p; it was proposed that precursor mRNA is also translated but the putative product of 230 aa is rapidly degraded by the ubiquitin proteasome pathway. We have identified and characterized the same regulated splicing and confirmed its essential features. Contrary to the above proposal, however, we find that the 238-aa product of mature mRNA and the 230-aa-type protein tested are highly unstable with little of no difference in stability. Furthermore, we demonstrate that the absence of Hac1p in unstressed cells is due to the lack of translation of precursor mRNA. We conclude that Hac1p is synthesized as the result of ER stress-induced mRNA splicing, leading to activation of the UPR. PMID- 9348530 TI - A novel yeast screen for mitotic arrest mutants identifies DOC1, a new gene involved in cyclin proteolysis. AB - B-type cyclins are rapidly degraded at the transition between metaphase and anaphase and their ubiquitin-mediated proteolysis is required for cells to exit mitosis. We used a novel enrichment to isolate new budding mutants that arrest the cell cycle in mitosis. Most of these mutants lie in the CDC16, CDC23, and CDC27 genes, which have already been shown to play a role in cyclin proteolysis and encode components of a 20S complex (called the cyclosome or anaphase promoting complex) that ubiquitinates mitotic cyclins. We show that mutations in CDC26 and a novel gene, DOC1, also prevent mitotic cyclin proteolysis. Mutants in either gene arrest as large budded cells with high levels of the major mitotic cyclin (Clb2) protein at 37 degrees C and cannot degrade Clb2 in G1-arrested cells. Cdc26 associates in vivo with Doc1, Cdc16, Cdc23, and Cdc27. In addition, the majority of Doc1 cosediments at 20S with Cdc27 in a sucrose gradient, indicating that Cdc26 and Doc1 are components of the anaphase promoting complex. PMID- 9348529 TI - Modulation of in vivo migratory function of alpha 2 beta 1 integrin in mouse liver. AB - We report herein that expression of alpha 2 beta 1 integrin increased human erythroleukemia K562 transfectant (KX2C2) cell movement after extravasation into liver parenchyma. In contrast, a previous study demonstrated that alpha 2 beta 1 expression conferred a stationary phenotype to human rhabdomyosarcoma RD transfectant (RDX2C2) cells after extravasation into the liver. We therefore assessed the adhesive and migratory function of alpha 2 beta 1 on KX2C2 and RDX2C2 cells using a alpha 2 beta 1-specific stimulatory monoclonal antibody (mAb), JBS2, and a blocking mAb, BHA2.1. In comparison with RDX2C2 cells, KX2C2 were only weakly adherent to collagen and laminin. JBS2 stimulated alpha 2 beta 1 mediated interaction of KX2C2 cells with both collagen and laminin resulting in increases in cell movement on both matrix proteins. In the presence of Mn2+, JBS2 stimulated adhesion on collagen beyond an optimal level for cell movement. In comparison, an increase in RDX2C2 cell movement on collagen required a reduction in its adhesive strength provided by the blocking mAb BHA2.1. Consistent with these in vitro findings, in vivo videomicroscopy revealed that alpha 2 beta 1 mediated postextravasation cell movement of KX2C2 cells in the liver tissue could also be stimulated by JBS2. Thus, results demonstrate that alpha 2 beta 1 expression can modulate postextravasation cell movement by conferring either a stationary or motile phenotype to different cell types. These findings may be related to the differing metastatic activities of different tumor cell types. PMID- 9348531 TI - 14-3-3 inhibits the Dictyostelium myosin II heavy-chain-specific protein kinase C activity by a direct interaction: identification of the 14-3-3 binding domain. AB - Myosin II heavy chain (MHC) specific protein kinase C (MHC-PKC), isolated from Dictyostelium discoideum, regulates myosin II assembly and localization in response to the chemoattractant cyclic AMP. Immunoprecipitation of MHC-PKC revealed that it resides as a complex with several proteins. We show herein that one of these proteins is a homologue of the 14-3-3 protein (Dd14-3-3). This protein has recently been implicated in the regulation of intracellular signaling pathways via its interaction with several signaling proteins, such as PKC and Raf 1 kinase. We demonstrate that the mammalian 14-3-3 zeta isoform inhibits the MHC PKC activity in vitro and that this inhibition is carried out by a direct interaction between the two proteins. Furthermore, we found that the cytosolic MHC-PKC, which is inactive, formed a complex with Dd14-3-3 in the cytosol in a cyclic AMP-dependent manner, whereas the membrane-bound active MHC-PKC was not found in a complex with Dd14-3-3. This suggests that Dd14-3-3 inhibits the MHC PKC in vivo. We further show that MHC-PKC binds Dd14-3-3 as well as 14-3-3 zeta through its C1 domain, and the interaction between these two proteins does not involve a peptide containing phosphoserine as was found for Raf-1 kinase. Our experiments thus show an in vivo function for a member of the 14-3-3 family and demonstrate that MHC-PKC interacts directly with Dd14-3-3 and 14-3-3 zeta through its C1 domain both in vitro and in vivo, resulting in the inhibition of the kinase. PMID- 9348532 TI - Interaction of coatomer with aminoglycoside antibiotics: evidence that coatomer has at least two dilysine binding sites. AB - Coatomer is the soluble precursor of the COPI coat (coat protein I) involved in traffic among membranes of the endoplasmic reticulum and the Golgi apparatus. We report herein that neomycin precipitates coatomer from cell extracts and from purified coatomer preparations. Precipitation first increased and then decreased as the neomycin concentration increased, analogous to the precipitation of a polyvalent antigen by divalent antibodies. This suggested that neomycin cross linked coatomer into large aggregates and implies that coatomer has two or more binding sites for neomycin. A variety of other aminoglycoside antibiotics precipitated coatomer, or if they did not precipitate, they interfered with the ability of neomycin to precipitate. Coatomer is know to interact with a motif (KKXX) containing adjacent lysine residues at the carboxyl terminus of the cytoplasmic domains of some membrane proteins resident in the endoplasmic reticulum. All of the antibiotics that interacted with coatomer contain at least two close amino groups, suggesting that the antibiotics might be interacting with the di-lysine binding site of coatomer. Consistent with this idea, di-lysine itself blocked the interaction of antibiotics with coatomer. Moreover, di-lysine and antibiotics each blocked the coating of Golgi membranes by coatomer. These data suggest that certain aminoglycoside antibiotics interact with di-lysine binding sites on coatomer and that coatomer contains at least two of these di lysine binding sites. PMID- 9348533 TI - Characterization of the Golgi complex cleared of proteins in transit and examination of calcium uptake activities. AB - To characterize endogenous molecules and activities of the Golgi complex, proteins in transit were > 99% cleared from rat hepatocytes by using cycloheximide (CHX) treatment. The loss of proteins in transit resulted in condensation of the Golgi cisternae and stacks. Isolation of a stacked Golgi fraction is equally efficient with or without proteins in transit [control (CTL SGF1) and cycloheximide (CHX SGF1)]. Electron microscopy and morphometric analysis showed that > 90% of the elements could be positively identified as Golgi stacks or cisternae. Biochemical analysis showed that the cis-, medial-, trans-, and TGN Golgi markers were enriched over the postnuclear supernatant 200- to 400-fold with and 400- to 700-fold without proteins in transit. To provide information on a mechanism for import of calcium required at the later stages of the secretory pathway, calcium uptake into CTL SGF1 and CHX SGF1 was examined. All calcium uptake into CTL SGF1 was dependent on a thapsigargin-resistant pump not resident to the Golgi complex and a thapsigargin-sensitive pump resident to the Golgi. Experiments using CHX SGF1 showed that the thapsigargin-resistant activity was a plasma membrane calcium ATPase isoform in transit to the plasma membrane and the thapsigargin-sensitive pump was a sarcoplasmic/endoplasmic reticulum calcium ATPase isoform. In vivo both of these calcium ATPases function to maintain millimolar levels of calcium within the Golgi lumen. PMID- 9348534 TI - Segregation of two spectrin isoforms: polarized membrane-binding sites direct polarized membrane skeleton assembly. AB - Spectrin isoforms are often segregated within specialized plasma membrane subdomains where they are thought to contribute to the development of cell surface polarity. It was previously shown that ankyrin and beta spectrin are recruited to sites of cell-cell contact in Drosophila S2 cells expressing the homophilic adhesion molecule neuroglian. Here, we show that neuroglian has no apparent effect on a second spectrin isoform (alpha beta H), which is constitutively associated with the plasma membrane in S2 cells. Another membrane marker, the Na,K-ATPase, codistributes with ankyrin and alpha beta spectrin at sites of neuroglian-mediated contact. The distributions of these markers in epithelial cells in vivo are consistent with the order of events observed in S2 cells. Neuroglian, ankyrin, alpha beta spectrin, and the Na,K-ATPase colocalize at the lateral domain of salivary gland cells. In contrast, alpha beta H spectrin is sorted to the apical domain of salivary gland and somatic follicle cells. Thus, the two spectrin isoforms respond independently to positional cues at the cell surface: in one case an apically sorted receptor and in the other case a locally activated cell-cell adhesion molecule. The results support a model in which the membrane skeleton behaves as a transducer of positional information within cells. PMID- 9348536 TI - The balance of RanBP1 and RCC1 is critical for nuclear assembly and nuclear transport. AB - Ran is a small GTPase that is essential for nuclear transport, mRNA processing, maintenance of structural integrity of nuclei, and cell cycle control. RanBP1 is a highly conserved Ran guanine nucleotide dissociation inhibitor. We sought to use Xenopus egg extracts for the development of an in vitro assay for RanBP1 activity in nuclear assembly, protein import, and DNA replication. Surprisingly, when we used anti-RanBP1 antibodies to immunodeplete RanBP1 from Xenopus egg extracts, we found that the extracts were also depleted of RCC1, Ran's guanine nucleotide exchange factor, suggesting that these proteins form a stable complex. In contrast to previous observations using extracts that had been depleted of RCC1 only, extracts lacking both RanBP1 and RCC1 (codepleted extracts) did not exhibit defects in assays of nuclear assembly, nuclear transport, or DNA replication. Addition of either recombinant RanBP1 or RCC1 to codepleted extracts to restore only one of the depleted proteins caused abnormal nuclear assembly and inhibited nuclear transport and DNA replication in a manner that could be rescued be further addition of RCC1 or RanBP1, respectively. Exogenous mutant Ran proteins could partially rescue nuclear function in extracts without RanBP1 or without RCC1, in a manner that was correlated with their nucleotide binding state. These results suggest that little RanBP1 or RCC1 is required for nuclear assembly, nuclear import, or DNA replication in the absence of the other protein. The results further suggest that the balance of GTP- and GDP-Ran is critical for proper nuclear assembly and function in vitro. PMID- 9348535 TI - Quality control in the secretory pathway: the role of calreticulin, calnexin and BiP in the retention of glycoproteins with C-terminal truncations. AB - Unlike properly folded and assembled proteins, most misfolded and incompletely assembled proteins are retained in the endoplasmic reticulum of mammalian cells and degraded without transport to the Golgi complex. To analyze the mechanisms underlying this unique sorting process and its fidelity, the fate of C-terminally truncated fragments of influenza hemagglutinin was determined. An assortment of different fragments was generated by adding puromycin at low concentrations to influenza virus-infected tissue culture cells. Of the fragments generated, < 2% was secreted, indicating that the system for detecting defects in newly synthesized proteins is quite stringent. The majority of secreted species corresponded to folding domains within the viral spike glycoprotein. The retained fragments acquired a partially folded structure with intra-chain disulfide bonds and conformation-dependent antigenic epitopes. They associated with two lectin like endoplasmic reticulum chaperones (calnexin and calreticulin) but not BiP/GRP78. Inhibition of the association with calnexin and calreticulin by the addition of castanospermine significantly increased fragment secretion. However, it also caused association with BiP/GRP78. These results indicated that the association with calnexin and calreticulin was involved in retaining the fragments. They also suggested that BiP/GRP78 could serve as a backup for calnexin and calreticulin in retaining the fragments. In summary, the results showed that the quality control system in the secretory pathway was efficient and sensitive to folding defects, and that it involved multiple interactions with endoplasmic reticulum chaperones. PMID- 9348537 TI - Subcellular localization of myosin-V in the B16 melanoma cells, a wild-type cell line for the dilute gene. AB - The discovery that the dilute gene encodes a class V myosin led to the hypothesis that this molecular motor is involved in melanosome transport and/or dendrite outgrowth in mammalian melanocytes. The present studies were undertaken to gain insight into the subcellular distribution of myosin-V in the melanoma cell line B16-F10, which is wild-type for the dilute gene. Immunofluorescence studies showed some degree of superimposed labeling of myosin-V with melanosomes that predominated at the cell periphery. A subcellular fraction highly enriched in melanosomes was also enriched in myosin-V based on Western blot analysis. Immunoelectron microscopy showed myosin-V labeling associated with melanosomes and other organelles. The stimulation of B16 cells with the alpha-melanocyte stimulating hormone led to a significant increase in myosin-V expression. This is the first evidence that a cAMP signaling pathway might regulate the dilute gene expression. Immunofluorescence also showed an intense labeling of myosin-V independent of melanosomes that was observed within the dendrites and at the perinuclear region. Although the results presented herein are consistent with the hypothesis that myosin-V might act as a motor for melanosome translocation, they also suggest a broader cytoplasmic function for myosin-V, acting on other types of organelles or in cytoskeletal dynamics. PMID- 9348538 TI - A ubiquitin-conjugating enzyme is essential for developmental transitions in Dictyostelium. AB - We have identified a developmentally essential gene, UbcB, by insertional mutagenesis. The encoded protein (UBC1) shows very high amino acid sequence identity to ubiquitin-conjugating enzymes from other organisms, suggesting that UBC1 is involved in protein ubiquitination and possibly degradation during Dictyostelium development. Consistent with the homology of the UBC1 protein to UBCs, the developmental pattern of protein ubiquitination is altered in ubcB-null cells. ubcB-null cells are blocked in the ability to properly execute the developmental transition that occurs between the induction of postaggregative gene expression during mound formation and the induction of cell-type differentiation and subsequent morphogenesis. ubcB-null cells plated on agar form mounds with normal kinetics; however, they remain at this stage for approximately 10 h before forming multiple tips and fingers that then arrest. Under other conditions, some of the fingers form migrating slugs, but no culmination is observed. In ubcB-null cells, postaggregative gene transcripts accumulate to very high levels and do not decrease significantly with time as they do in wild-type cells. Expression of cell-type-specific genes is very delayed, with the level of prespore-specific gene expression being significantly reduced compared with that in wild-type cells. lacZ reporter studies using developmentally regulated and cell-type-specific promoters suggest that ubcB-null cells show an unusually elevated level of staining of lacZ reporters expressed in anterior-like cells, a regulatory cell population found scattered throughout the aggregate, and reduced staining of a prespore reporter. ubcB-null cells in a chimeric organism containing predominantly wild-type cells are able to undergo terminal differentiation but show altered spatial localization. In contrast, in chimeras containing only a small fraction of wild-type cells, the mature fruiting body is very small and composed almost exclusively of wild-type cells, with the ubcB-null cells being present as a mass of cells located in extreme posterior of the developing organism. The amino acid sequence analysis of the UbcB open reading frame (ORF) and the analysis of the developmental phenotypes suggest that tip formation and subsequent development requires specific protein ubiquitination, and possibly degradation. PMID- 9348539 TI - Amphiphysin heterodimers: potential role in clathrin-mediated endocytosis. AB - Amphiphysin (Amph) is a src homology 3 domain-containing protein that has been implicated in synaptic vesicle endocytosis as a result of its interaction with dynamin. In a screen for novel members of the amphiphysin family, we identified Amph2, an isoform 49% identical to the previously characterized Amph1 protein. The subcellular distribution of this isoform parallels Amph1, both being enriched in nerve terminals. Like Amph1, a role in endocytosis at the nerve terminal is supported by the rapid dephosphorylation of Amph2 on depolarization. Importantly, the two isoforms can be coimmunoprecipitated from the brain as an equimolar complex, suggesting that the two isoforms act in concert. As determined by cross linking of brain extracts, the Amph1-Amph2 complex is a 220- to 250-kDa heterodimer. COS cells transfected with either Amph1 or Amph2 show greatly reduced transferrin uptake, but coexpression of the two proteins rescues this defect, supporting a role for the heterodimer in clathrin-mediated endocytosis. Although the src homology 3 domains of both isoforms interact with dynamin, the heterodimer can associate with multiple dynamin molecules in vitro and activates dynamin's GTPase activity. We propose that it is an amphiphysin heterodimer that drives the recruitment of dynamin to clathrin-coated pits in endocytosing nerve terminals. PMID- 9348540 TI - Nup93, a vertebrate homologue of yeast Nic96p, forms a complex with a novel 205 kDa protein and is required for correct nuclear pore assembly. AB - Yeast and vertebrate nuclear pores display significant morphological similarity by electron microscopy, but sequence similarity between the respective proteins has been more difficult to observe. Herein we have identified a vertebrate nucleoporin, Nup93, in both human and Xenopus that has proved to be an evolutionarily related homologue of the yeast nucleoporin Nic96p. Polyclonal antiserum to human Nup93 detects corresponding proteins in human, rat, and Xenopus cells. Immunofluorescence and immunoelectron microscopy localize vertebrate Nup93 at the nuclear basket and at or near the nuclear entry to the gated channel of the pore. Immunoprecipitation from both mammalian and Xenopus cell extracts indicates that a small fraction of Nup93 physically interacts with the nucleoporin p62, just as yeast Nic96p interacts with the yeast p62 homologue. However, a large fraction of vertebrate Nup93 is extracted from pores and is also present in Xenopus egg extracts in complex with a newly discovered 205-kDa protein. Mass spectrometric sequencing of the human 205-kDa protein reveals that this protein is encoded by an open reading frame, KIAAO225, present in the human database. The putative human nucleoporin of 205 kDa has related sequence homologues in Caenorhabditis elegans and Saccharomyces cerevisiae. The analyze the role of the Nup93 complex in the pore, nuclei were assembled that lack the Nup93 complex after immunodepletion of a Xenopus nuclear reconstitution extract. The Nup93-complex-depleted nuclei are clearly defective for correct nuclear pore assembly. From these experiments, we conclude that the vertebrate and yeast pore have significant homology in their functionally important cores and that, with the identification of Nup93 and the 205-kDa protein, we have extended the knowledge of the nearest-neighbor interactions of this core in both yeast and vertebrates. PMID- 9348541 TI - The rat myosin myr 5 is a GTPase-activating protein for Rho in vivo: essential role of arginine 1695. AB - myr 5 is an unconventional myosin (class IX) from rat that contains a Rho-family GTPase-activating protein (GAP) domain. Herein we addressed the specificity of the myr 5 GAP activity, the molecular mechanism by which GAPs activate GTP hydrolysis, the consequences of myr 5 overexpression in living cells, and its subcellular localization. The myr 5 GAP activity exhibits a high specificity for Rho. To achieve similar rates of GTPase activation for RhoA, Cdc42Hs, and Rac1, a 100-fold or 1000-fold higher concentration of recombinant myr 5 GAP domain was needed for Cdc42Hs or Rac1, respectively, as compared with RhoA. Cell lysates from Sf9 insect cells infected with recombinant baculovirus encoding myr 5 exhibited increased GAP activity for RhoA but not for Cdc42Hs or Rac1. Analysis of Rho-family GAP domain sequences for conserved arginine residues that might contribute to accelerate GTP hydrolysis revealed a single conserved arginine residue. Mutation of the corresponding arginine residue in the myr 5 GAP domain to a methionine (M1695) virtually abolished Rho-GAP activity. Expression of myr 5 in Sf9 insect cells induced the formation of numerous long thin processes containing occasional varicosities. Such morphological changes were dependent on the myr 5 Rho-GAP activity, because they were induced by expression the myr 5 tail or just the myr 5 Rho-GAP domain but not by expressing the myr 5 myosin domain. Expression of myr 5 in mammalian normal rat kidney (NRK) or HtTA-1 HeLa cells induced a loss of actin stress fibers and focal contacts with concomitant morphological changes and rounding up of the cells. Similar morphological changes were observed in HtTA-1 HeLa cells expressing just the myr 5 Rho-GAP domain but not in cells expressing myr 5 M1695. These morphological changes induced by myr 5 were inhibited by coexpression of RhoV14, which is defective in GTP hydrolysis, but not by RhoI117. myr 5 was localized in dynamic regions of the cell periphery, in the perinuclear region in the Golgi area, along stress fibers, and in the cytosol. These results demonstrate that myr 5 has in vitro and in vivo Rho-GAP activity. No evidence for a Rho effector function of the myr 5 myosin domain was obtained. PMID- 9348542 TI - Cell-adhesive responses to tenascin-C splice variants involve formation of fascin microspikes. AB - Tenascin-C is an adhesion-modulating matrix glycoprotein that has multiple effects on cell behavior. Tenascin-C transcripts are expressed in motile cells and at sites of tissue modeling during development, and alternative splicing generates variants that encode different numbers of fibronectin type III repeats. We have examined the in vivo expression and cell adhesive properties of two full length recombinant tenascin-C proteins: TN-190, which contains the eight constant fibronectin type III repeats, and TN-ADC, which contains the additional AD2, AD1, and C repeats. In situ hybridization with probes specific for the AD2, AD1, and C repeats shows that these splice variants are expressed at sites of active tissue modeling and fibronectin expression in the developing avian feather bud and sternum. Transcripts incorporating the AD2, AD1, and C repeats are present in embryonic day 10 wing bud but not in embryonic day 10 lung. By using a panel of nine cell lines in attachment assays, we have found that C2C12, G8, and S27 myoblastic cells undergo concentration-dependent adhesion to both variants, organize actin microspikes that contain the actin-bundling protein fascin, and do not assemble focal contacts. On a molar basis, TN-ADC is more active than TN-190 in promoting cell attachment and irregular cell spreading. The addition of either TN-190 or TN-ADC in solution to C2C12, COS-7, or MG-63 cells adherent on fibronectin decreases cell attachment and results in decreased organization of actin microfilament bundles, with formation of cortical membrane ruffles and retention of residual points of substratum contact that contain filamentous actin and fascin. These data establish a biochemical similarity in the processes of cell adhesion to tenascin-C and thrombospondin-1, also an "antiadhesive" matrix component, and also demonstrate that both the adhesive and adhesion-modulating properties of tenascin-C involve similar biochemical events in the cortical cytoskeleton. In addition to these generic properties, TN-ADC is less active in adhesion modulation than TN-190. The coordinated expression of different tenascin C transcripts during development may, therefore, provide appropriate microenvironments for regulated changes in cell shape, adhesion, and movement. PMID- 9348543 TI - The involvement of the intermediate chain of cytoplasmic dynein in binding the motor complex to membranous organelles of Xenopus oocytes. AB - Cytoplasmic dynein is one of the major motor proteins involved in intracellular transport. It is a protein complex consisting of four subunit classes: heavy chains, intermediate chains (ICs), light intermediate chains, and light chains. In a previous study, we had generated new monoclonal antibodies to the ICs and mapped the ICs to the base of the motor. Because the ICs have been implicated in targeting the motor to cargo, we tested whether these new antibodies to the intermediate chain could block the function of cytoplasmic dynein. When cytoplasmic extracts of Xenopus oocytes were incubated with either one of the monoclonal antibodies (m74-1, m74-2), neither organelle movement nor network formation was observed. Network formation and membrane transport was blocked at an antibody concentration as low as 15 micrograms/ml. In contrast to these observations, no effect was observed on organelle movement and tubular network formation in the presence of a control antibody at concentrations as high as 0.5 mg/ml. After incubating cytoplasmic extracts or isolated membranes with the monoclonal antibodies m74-1 and m74-2, the dynein IC polypeptide was no longer detectable in the membrane fraction by SDS-PAGE immunoblot, indicating a loss of cytoplasmic dynein from the membrane. We used a panel of dynein IC truncation mutants and mapped the epitopes of both antibodies to the N-terminal coiled-coil domain, in close proximity to the p150Glued binding domain. In an IC affinity column binding assay, both antibodies inhibited the IC-p150Glued interaction. Thus these findings demonstrate that direct IC-p150Glued interaction is required for the proper attachment of cytoplasmic dynein to membranes. PMID- 9348545 TI - In vivo imaging of neurotransmitter systems using radiolabeled receptor ligands. AB - In vivo functional brain imaging, including global blood flow, regional cerebral blood flow (rCBF), measured with positron emission tomography (PET) and single photon emission computed tomography (SPECT), and regional cerebral metabolic rate (rCMR) measured with deoxyglucose PET, have been widely used in studies of psychiatric disorders. These studies have found modest differences and required large numbers of patients. Activation studies using rCBF or rCMR as indices of neuronal activity are more sensitive because patients act as their own control; however, findings localize regions of change but provide no data about specific neurotransmitter systems. After a general discussion of the role of neurotransmitter systems in neuropsychiatric disorders, an overview of the methodology of development and selection of radioligands for PET and SPECT is presented. Studies involving PET and SPECT ligand methods are reviewed and their findings summarized, including recent work demonstrating successive mutual modulation of neurotransmitter systems. Kinetic and equilibrium analysis modeling are reviewed. The emerging methodology of measuring neurotransmitter release on activation, both pharmacologically and by task performance, using ligand methods is reviewed and proposed as a promising new approach for studying psychiatric disorders. PMID- 9348544 TI - Transport of myosin II to the equatorial region without its own motor activity in mitotic Dictyostelium cells. AB - Fluorescently labeled myosin moved and accumulated circumferentially in the equatorial region of dividing Dictyostelium cells within a time course of 4 min, followed by contraction of the contractile ring. To investigate the mechanism of this transport process, we have expressed three mutant myosins that cannot hydrolyze ATP in myosin null cells. Immunofluorescence staining showed that these mutant myosins were also correctly transported to the equatorial region, although no contraction followed. The rates of transport, measured using green fluorescent protein-fused myosins, were indistinguishable between wild-type and mutant myosins. These observations demonstrate that myosin is passively transported toward the equatorial region and incorporated into the forming contractile ring without its own motor activity. PMID- 9348546 TI - Autoradiographic localization of CRF1 and CRF2 binding sites in adult rat brain. AB - The regional distribution of corticotropin-releasing factor1 (CRF1) and CRF2 binding sites was assessed autoradiographically in adult rat brain. The differential pharmacological profiles of the CRF1 and CRF2 receptor subtypes were used for the discrimination of the CRF1 and CRF2 receptor subtypes in rat brain. Pharmacological characterization at the human CRF1 receptor subtype, expressed in baculovirus-infected Sf9 cells, showed high affinity binding (Ki < or = 10.0 nM) for the peptide agonists sauvagine, urotensin I, rat/human CRF, and ovine CRF. Pharmacological characterization at the rat CRF2 receptor subtype expressed in CHO cells showed a rank order affinity for the peptide agonists such that sauvagine, urotensin I and rat/human CRF showed high affinity binding whereas ovine CRF had a Ki value of 300 nM. Based on this differential binding affinity for ovine CRF, [125I]sauvagine binding in the presence of increasing concentrations of ovine CRF was used to discriminate CRF1 from CRF2 receptor subtypes in rat brain. The CRF1 receptor subtype was found to be localized to various regions of the cerebellum, as well as to several cortical areas. The CRF2 receptor subtype was shown to be localized to the lateral septal nucleus, entorhinal cortex, and to amygdaloid and hypothalamic regions. The present autoradiographic findings provide evidence that each subtype has a distinct regional distribution, thus strengthening the suggestion that CRF1 and CRF2 receptors serve different roles in mediating CRF function. Such data suggest that the development of CRF receptor subtype selective antagonists should help to delineate the role of CRF1 and CRF2 receptor subtypes in central nervous system function. PMID- 9348547 TI - Effects of clozapine on plasma catecholamines and relation to treatment response in schizophrenia: a within-subject comparison with haloperidol. AB - We conducted a within-subject comparison of the effects of clozapine and haloperidol on plasma levels of neurotransmitters and metabolites, and related changes in specific plasma neurochemicals with clozapine response. The subjects were 14 inpatients with schizophrenia or schzoaffective disorder, who were refractory to haloperidol and at least one other typical antipsychotic medication. Subjects underwent, in the following order: a 6-week "fixed, flexible dose" haloperidol trial, followed by a 2-4 week medication-free phase, and a 6 week clozapine trial. Plasma levels of norepinephrine (NE), homovanillic acid (HVA), and 3-methoxy-4-hydroxyphenylglycol (MHPG), and objective clinical ratings of total, positive, negative, and depressive symptoms were obtained at the end of each phase. As expected, we found a substantial increase of plasma NE with clozapine but not with haloperidol. However, the increase in NE was not associated with improvement in total or positive symptomatology. There was some evidence for an association between improvement in negative symptoms and increased HVA on clozapine, as well as diminished HVA during the medication-free phase. The implications of these data for understanding the mechanisms of action of clozapine are discussed. PMID- 9348548 TI - Differential distribution of corticotropin-releasing hormone immunoreactive axons in monoaminergic nuclei of the human brainstem. AB - Corticotropin-releasing hormone (CRH) has been implicated in a variety of physiological and behavioral responses to stress, as well as in the pathophysiology of certain psychiatric disorders. Although studies in rodents support a neuromodulatory influence of CRH on monoamine neurotransmission in a number of brain regions, little information in available to support a similar role for CRH in the human brain. The present study used immunocytochemistry to characterize the anatomical organization of CRH-immunoreactive axons in the human brainstem. Substantial regional differences in the density and distribution of CRH-immunoreactive axons were found in the dopamine-, noradrenaline- and serotonin-containing cell body regions of the human brainstem. Dense networks of CRH-immunoreactive axons were found in the medial subnuclei of the ventral mesencephalon and in the dorsolateral region of the locus coeruleus. Moderate densities of CRH-positive fibers were located in the median and dorsal raphe, whereas lower numbers of CRH-labeled axons appeared in the substantia nigra pars compacta. In addition, differences in CRH innervation density were observed within each region. For example, the dorsal tier of the substantia nigra contained a greater density of CRH-labeled axons than the ventral tier. In all monoamine-containing nuclei, CRH-labeled axons exhibited numerous beaded varicosities and fine intervaricose segments. The differential distribution of CRH-containing axons across these human brainstem nuclei suggests that the influence of CRH on monoamine function may be neurotransmitter-specific. PMID- 9348550 TI - More comments on hepatic artery infusion. PMID- 9348549 TI - Neurobiology of tryptophan depletion in depression: effects of m chlorophenylpiperazine (mCPP). AB - This study utilized neuroendocrine and mood responses to intravenous (i.v.) infusion of the serotonin (5-HT) agonist m-chlorophenylpiperazine (mCPP) to evaluate central 5-HT function in depressed patients undergoing acute tryptophan (TRP) depletion. Twenty-two drug-free patients with DSM-III-R major depression participated. Each patient underwent two randomized, double-blind TRP depletion tests, one sham and one active. At the estimated time of maximum TRP depletion, each patient received an i.v. infusion of mCPP 0.1 mg/kg. Blood was obtained for serum cortisol, prolactin, and growth hormone. Multiple rating scales were used to assess mood. The cortisol response to i.v. mCPP was significantly greater during TRP depletion than during sham depletion, and free plasma TRP was negatively correlated with the cortisol response during TRP depletion. These findings are consistent with the hypothesis that acute TRP depletion in drug-free depressed patients induces a compensatory up-regulation of postsynaptic 5-HT receptors, most likely of the 5-HT2A/2C subtype. Such changes suggest a mechanism by which acute and potent manipulations of 5-HT function in depressed patients could be used to effect rapid clinical improvement. PMID- 9348551 TI - Is hepatic artery chemotherapy for everyone? PMID- 9348552 TI - New procedures recommended for improvement of surgical blood transfusion. PMID- 9348553 TI - Coalition president urges policy board to focus on the policy and practices affecting cancer research. PMID- 9348554 TI - Is axillary dissection always indicated in invasive breast cancer? AB - In light of the changing trends in the diagnosis and management of invasive breast cancer, the practice of routine axillary dissection should be reevaluated. A growing number of patients with breast cancer are diagnosed as having small tumors with an associated low risk of lymph node metastases. The pathologic features of the primary tumor are increasingly being used as a prognostic guide for recommendations about adjuvant systemic therapy, and there are recent reports suggesting a superior prognostic value for tumor cells detected in bone marrow, as compared to axillary lymph node metastases. Consequently, axillary lymph node status is no longer the single prognostic guide for recommendations about adjuvant systemic therapy. For treatment of the axilla, there is evidence that, in clinical N0 patients, radiation therapy to the axilla is an effective alternative to axillary dissection. Finally, there are cost and morbidity considerations for patients undergoing axillary dissection in whom the indications of the procedure are equivocal. In the management of invasive breast cancer, a selective policy toward axillary lymph node dissection should be considered. This review discusses the nonsurgical management of the axilla; ie, radiation therapy to the axilla and observation of the axilla as an alternative to axillary dissection. PMID- 9348555 TI - Estrogen replacement therapy for breast cancer patients. AB - Female reproductive hormones cause breast cancer. Long-term use of postmenopausal hormones increases the risk of breast cancer. The apparent survival advantage seen in women diagnosed with breast cancer while taking postmenopausal hormones may be due to the hormone-responsive nature of their tumors, diagnosis at an earlier stage, or other biases. Thus, the data indicating a survival advantage do not specify what the policies were on the use of hormones after diagnosis. Substantial evidence from studies of obesity confirms the association of higher estrogen levels with poorer prognosis. Tamoxifen (Nolvadex), acting as an antiestrogen in breast tissue, increases the likelihood of survival, as does oophorectomy in premenopausal women. Given these data, women diagnosed with breast cancer should use hormones sparingly. Alternatives to hormone therapy should be used for long-term prevention of heart disease and osteoporosis. PMID- 9348556 TI - The timing of breast cancer surgery during the menstrual cycle. AB - A number of recent studies have suggested that survival among premenopausal women after primary treatment of breast cancer may be affected by the estimated hormonal milieu at the time of surgery, especially in those with axillary lymph node metastases. The concept has created considerable controversy and has resulted in the publication of many negative reports. However, several biological mechanisms have been suggested for the observed survival advantage. These include cyclical patterns of immune function, as well as cell division and cell death, that correlate with hormonal fluctuations of the menstrual cycle. Comparisons among studies of timing have been complicated by differences in menstrual cycle divisions, variability in the sources of study populations, limited availability of menstrual history data, and changes over the past 2 decades in primary and adjuvant breast cancer therapy. Several recent publications have been enhanced by the availability of serum collected at the time of surgery that enables accurate measurement of the hormonal milieu. In these studies, the likelihood of misclassification by menstrual cycle phase is reduced, and dependence on recalled menstrual history is eliminated. High progesterone levels have been associated with improved survival. These findings have encouraged some to suggest that perioperative administration of progesterone or tamoxifen (Nolvadex) may provide a preventive avenue comparable to scheduling surgery during the luteal phase. Further multidisciplinary studies are needed, however, to clarify the influence of the naturally occurring or medically induced hormonal milieu at the time of breast cancer surgery on survival in premenopausal women. PMID- 9348557 TI - Clinical trials referral resource. Clinical trials in head and neck cancer. PMID- 9348558 TI - The economics of prostate cancer screening. AB - The introduction of prostate-specific antigen (PSA) testing for use in the early detection of prostate cancer has led to controversy regarding the appropriateness of prostate cancer screening and any subsequent treatment. Much of this controversy arises from concern over the increased health-care costs that may result from widespread screening. As cost control becomes a dominant concern in today's health-care system, practitioners must decide whether the expense of screening and resulting treatment is worth the expenditure of its limited health care system, practitioners must decide whether the expense of screening and resulting treatment is worth the expenditure of its limited health-care resources. This review first discusses the effects that widespread PSA screening would have on health-care costs. The benefits that will be realized by the expenditure of these additional health-care dollars are much more difficult to quantify. Decision analysis models have been used to evaluate the effectiveness of prostate cancer screening and treatment and have found little or no benefit. The current review illustrates how assumptions used to construct these models influence their results. The authors present a quantitative analysis of the costs and benefits of prostate cancer screening and treatment. This type of analysis demonstrates that prostate cancer screening and treatment may be a very cost effective health-care intervention. Although men 50 to 70 years old will potentially benefit the most from PSA screening, this benefit will not be realized until they are in their seventh or eight decade of life. Society must decide if the years of life saved in these men warrants the use of its limited health-care resources. This decision will be easier when randomized, controlled trials are available to quantify the costs and benefits of PSA screening. PMID- 9348559 TI - Management of progressive metastatic prostate cancer. AB - Metastatic prostate cancer is a growing health problem and is the second leading cause of cancer death in men. While the response of patients with metastatic prostate cancer to initial hormonal manipulation is excellent, the majority of patients eventually progress. As a result, a growing number of patients and their physicians need-to-find acceptable therapeutic alternatives. Fortunately, the number of therapies in the management armamentarium is growing and includes: alternative hormonal therapies, chemotherapy, radioisotopes, and investigational agents. The major focus of treatment has shifted to palliation and quality of life. The decline of prostate-specific antigen (PSA) has become another important end point as evidence supporting a correlation with prolonged survival mounts. Enrolling eligible patients in clinical trials is critical to the development of new treatment strategies for this difficult disease. PMID- 9348560 TI - UFT: East meets West in drug development. PMID- 9348561 TI - Scientific basis for the combination of tegafur with uracil. AB - Fujii et al reported that Uracil potentiated the antitumor activity of fluorouracil (5-FU) and 1-(2-tetrahydrofuryl)-5-fluorouracil (tegafur). This effect was due to inhibition of the degradation of 5-FU, yet the phosphorylation of 5-FU was unaffected. The molar ratio of tegafur and uracil was 1:4, a combination that has since been widely prescribed in Japan for the treatment of cancer patients. We present here our experimental and clinical results when investigating the antineoplastic effects of this combination of drugs--known as UFT--and provide evidence that UFT is an effective treatment for patients with cancer. PMID- 9348562 TI - Oral UFT plus leucovorin in patients with relapsed or refractory colorectal cancer. AB - Therapeutic options for patients with advanced colorectal cancer who have failed treatment with fluorouracil (5-FU) are limited. Responses have been reported in this setting with a protracted venous infusion of 5-FU. Daily oral therapy with tegafur and uracil (UFT) plus leucovorin (LV) has the potential to mimic the pharmacology of continuous infusion 5-FU. Therefore, we undertook a phase II study of a 28-day schedule of a combination chemotherapy regimen containing oral UFT/leucovorin in patients with measurable metastatic colorectal cancer who had failed treatment with bolus 5-FU. In addition, we sought to determine whether coadministration of UFT and leucovorin alters the bioavailability of these agents. In a pretreatment phase, each patient underwent sequential pharmacokinetic sampling following a single dose of UFT alone, leucovorin alone, and the combination of UFT plus leucovorin. The preliminary results of this trial suggest that tegafur pharmacokinetics are not affected by coadministration of leucovorin and that folate pharmacokinetics are not affected by UFT. PMID- 9348563 TI - Rationale for phase I study of UFT plus leucovorin and oral JM-216. AB - Both cisplatin (Platinol) and fluorouracil (5-FU) have demonstrated single-agent clinical efficacy in a variety of solid neoplasms. The combination of these agents has revealed synergistic cytotoxicity in models in vitro and in vivo, which may explain the clinical effectiveness of 5-FU-cisplatin regimens. UFT (tegafur and uracil) and bis-aceto-ammine-dichloro-cyclohexyl-amine platinum(IV)(JM-216) are novel oral analogues of 5-FU and cisplatin, respectively. In preclinical models, JM-216 has demonstrated equivalent cytotoxicity to cisplatin, while phase I trials suggest its dose-limiting toxicity is myelosuppression. In contrast to cisplatin, JM-216 has not demonstrated significant neurotoxicity or nephrotoxicity. UFT has been used extensively in Japan, where phase II data suggest disease response rates similar to single-agent 5-FU in colorectal, gastric, and breast carcinomas. Combination studies of prolonged administration UFT and single-dose cisplatin have shown efficacy, but also significant hematologic toxicity. We propose a phase I study of UFT and JM-216 administered daily over 14 consecutive days with leucovorin (90 mg/d). Ease of administration and continuous drug exposure are potential advantages of this regimen. Several disease specific investigations may be warranted given demonstrated feasibility in this phase I study. PMID- 9348564 TI - Experience with UFT in Japan. AB - The selective antineoplastic effect of tegafur and uracil (UFT) is attributed to its preferential enhancement of fluorouracil concentration in tumor tissues compared with that in normal tissues. The result of this effect is evident in the clinical benefit and lower toxicity associated with UFT compared with other fluorinated pyrimidines. Beginning with preclinical studies in the 1980s, significant therapeutic advantages of UFT have been reported in numerous trials conducted in Japan, including phase I dose-finding studies, phase II multicenter studies, comparative studies, and combination-chemotherapy studies. In phase II studies conducted at 211 institutions, for example, it was shown that the response rate was over 30% in patients with head/neck, bladder, or breast cancer, and the survival rate was superior to that previously reported in Japanese studies. Two comparative studies suggested that UFT was more effective than single-agent tegafur, and a number of combination-chemotherapy studies have shown that it has an advantage in terms of toxicity, response, and/or survival. UFT is also useful for postoperative adjuvant therapy, as well as therapy for advanced disease in a variety of neoplasms. UFT holds considerable promise and future trials should continue the evaluation and refinement of its role in the treatment of cancer. PMID- 9348565 TI - Phase I and pharmacokinetic evaluations of UFT plus oral leucovorin. AB - The phase I development program of tegafur and uracil (UFT) in the United States has included evaluation of the drug as a single agent and subsequent studies of its biochemical modulation by oral leucovorin. Phase I trials of single-agent UFT examined both a 5-day schedule repeated every 21 days and a 28-day schedule repeated every 35 days. In all of the trials the total dose was divided by three and administered three times daily at 8-hour intervals. Like intravenous schedules of fluorouracil (5-FU), UFT has schedule-dependent toxicity, with granulocytopenia being the dose-limiting toxicity for the 5-day regimen and diarrhea being the dose-limiting toxicity for the 28-day regimen. The suggested phase II doses for UFT administered without leucovorin were 800 mg/m2/day for the 5-day schedule and 360 mg/m2/day for the 28-day schedule. Subsequent phase I studies combining UFT with oral leucovorin used a 28-day schedule repeated every 35 days. Diarrhea was the dose-limiting toxicity, and the recommended phase II dose was UFT, 300 mg/m2/day, plus leucovorin, 90 mg/day. Pharmacokinetic evaluation showed that single-dose UFT results in maximum plasma levels and an area under the concentration-time curve that increased with escalating UFT doses. In addition, 5-FU levels were detectable throughout the 28-day dosing period; however, there was no evidence of significant accumulation of uracil, tegafur, or 5-FU. The administration of leucovorin in this trial provided continuous exposure of d,l-leucovorin and 5-methyltetrahydrofolate with little variation between doses or days. PMID- 9348566 TI - Postoperative adjuvant chemotherapy with mitomycin C and UFT for rectal cancer. AB - To evaluate the significance of postoperative adjuvant chemotherapy using mitomycin C (MMC) and UFT (tegafur and uracil) in combination, the Japanese Foundation for Multidisciplinary Treatment of Cancer conducted a prospective randomized controlled trial with 834 patients who had undergone curative resection for rectal cancer (T3 or T4 tumors and/or N1, N2, or N3 disease). The patients were randomly allocated to a treatment group (MMC/UFT, 416 patients) and a control group (surgery only, 418 patients). For patients in the treatment group, 20 mg of MMC was sprinkled on the operating field upon completion of surgery. MMC was intravenously injected at 6 mg/m2 on day 7, and then each month after surgery for 6 months. UFT was administered orally at 400 mg/day for 1 year. Although no difference was observed in the 5-year survival rate between the two groups, the 5-year disease-free survival rate in the MMC/UFT group was 69.1%, which was significantly higher than in the control group (59.3%, P = .005). The 5 year cumulative local recurrence rate was significantly lower in the MMC/UFT group (11.6%) than in the control group (19.0%) (P = .0071). We conclude that the adjuvant use of long-term oral UFT and intermittent intravenous MMC improves the disease-free survival rate of patients with curatively resected rectal cancer. PMID- 9348567 TI - Future directions in the adjuvant treatment of colon cancer. AB - Adjuvant chemotherapy has been shown to alter the natural history of patients with resected colon cancer. Two regimens (fluorouracil [5-FU] plus levamisole (Ergamisol) and 5-FU plus leucovorin) have been found most successful in prolonging disease-free and overall survival. When these two regimens were directly compared in randomized clinical trials, it appeared that a small disease free survival and overall survival advantage had emerged in favor of 5-FU plus leucovorin. This advantage, in conjunction with an increased understanding of the mechanism of leucovorin's biochemical modulation of 5-FU, makes this regimen a logical choice for studies designed to further optimize and augment the clinical efficacy of chemotherapy for colon cancer. The introduction of oral tegafur and uracil (UFT) and the demonstration of significant antitumor activity with the combination of oral UFT and oral leucovorin, provide an excellent opportunity to optimize treatment with 5-FU plus leucovorin. The National Surgical Adjuvant Breast and Bowel Project has recently implemented a new clinical trial (Protocol C-06) comparing oral UFT plus leucovorin with 5-FU plus leucovorin in the treatment of patients with resected stage II and III colon cancer. The rationale for the design of the trial, inclusion and exclusion criteria, treatment regimens, and statistical considerations are reviewed. PMID- 9348568 TI - 5-FU or UFT combined with leucovorin for previously untreated metastatic colorectal Ca. AB - This phase III study compares leucovorin plus fluorouracil (5-FU) 425 mg/m2, days 1 through 5, 28-day cycle, with oral leucovorin plus oral UFT (tegafur and uracil) 300 mg/m2, days 1 through 28, 35-day cycle, in terms of efficacy, safety, quality of life, and pharmacoeconomics. Eligible patients have not been treated previously and have measurable or evaluable metastatic colorectal cancer, an Eastern Cooperative Oncology Group performance status of 2 or less, and adequate bone marrow, liver, and renal functions. Patients are evaluated for response clinically and by computed tomography. Responses are determined by World Health Organization criteria. The study is nearing completion, with no toxicity issues requiring protocol modification. The results of this study could lead to a change to oral therapy as the standard of care for metastatic colorectal cancer, providing the efficacy and toxicity of UFT/leucovorin are at least equivalent due to the ease of administration and patient preference for oral regimens. PMID- 9348569 TI - UFT plus leucovorin vs 5-FU plus leucovorin for metastatic colorectal cancer. AB - An open-label, randomized phase III trial has been established to compare the efficacy and safety profile of tegafur and uracil (UFT) plus leucovorin with fluorouracil (5-FU) plus leucovorin as first-line chemotherapy for patients with metastatic colorectal adenocarcinoma. The primary end point of this study is time to progression. Secondary end points include tumor response, symptom control, quality of life, and pharmacoeconomics. Patients randomized to the experimental arm will receive UFT 300 mg/m2/day for 28 days followed by 1 week of rest, plus leucovorin 30 mg three times daily for 28 days. Patients in the control arm will receive 5-FU 425 mg/m2/day plus leucovorin 20 mg/m2/day for 5 days every 35 days. All patients eligible to participate in the study have evaluable or measurable disease. Assessments of tumor size and symptoms will be conducted every 5 weeks (every cycle), with scanning investigations repeated every 10 weeks (two cycles). Symptom evaluation includes assessment of pain, analgesic use, weight loss, and performance status. The intention was to recruit 362 patients by April 1996, with accrual planned for completion by July 1997. As of May 1997, 312 patients had been randomized in 15 countries, covering 45 active sites. All patients had good performance status, and the majority had no prior adjuvant chemotherapy. The study is currently ongoing, and no safety data are available at this time. PMID- 9348570 TI - A preliminary report of a phase II trial. UFT plus oral folinic acid as therapy for metastatic colorectal cancer in older patients. Spanish Group for the Treatment of Gastrointestinal Tumors (TTd Group). AB - The oral fluoropyrimidines have proved to be active in colorectal cancer in Japan and, recently, in the United States and Europe. Continuous oral administration simulates protracted fluorouracil (5-FU) continuous intravenous infusion. The purpose of this trial was to evaluate the tolerability and potential advantages of oral treatment for colorectal cancer in the elderly. The main inclusion criterion was age over 72 years. Patients were treated with UFT (tegafur plus uracil) 400 mg/24 hours (fixed doses) continuously plus folinic acid 45 mg/24 hours until toxicity. If grade 3 or 4 toxicity appeared, treatment was stopped until recovery. From September 1994 to November 1996, 126 patients were included. For the analysis in November 1996, 77 patients were evaluable for response, toxicity, and survival. The patients, including 34 women and 43 men, had a median age of 74 years (range, 72 to 82 years of age). The Karnofsky performance status was 60% to 80% for 41 patients and 90% to 100% for 36 patients. Liver metastasis was present in 48% of the cases, and 42% were locoregional and peritoneal. Toxicity was mild, with only one patient having grade 3 thrombocytopenia, 11 (14%) grade 3 or 4 nausea/vomiting, seven (9%) grade 3 or 4 diarrhea, and one grade 3 mucositis. Four patients (5%) had complete responses and nine (11.6%) partial responses, for an objective response rate of 16.9% (95% confidence interval, 9% to 27%). Twenty-two patients (28.6%) showed no change. The number of patients in whom disease did not progress (ie, patients with complete plus partial responses plus those with stable disease) was 35 (45.4%) (95% confidence interval, 34% to 57%). With a maximum follow-up of 24 months, the median actuarial survival is 14.4 months. The number without disease progression and the median survival in this preliminary analysis suggests that this schedule is a moderately effective, comfortable, treatment with only mild toxicity, that can be recommended for use in the elderly, and it warrants further study. PMID- 9348571 TI - Preoperative combined oral UFT plus leucovorin and radiation therapy for rectal cancer. AB - Several trials performed in the United States and Europe have demonstrated the efficacy of UFT (uracil and tegafur in a 4:1 molar combination) with oral leucovorin in the treatment of several tumor types, but particularly for advanced colorectal cancer. Phase III studies are under way in the United States to determine whether the combination of UFT with oral leucovorin is as effective as standard treatment, not only in the advanced setting but also in the adjuvant arena as well. This study is an open-label phase I trial to determine the safety of UFT and leucovorin, both given orally three times daily during concurrent fixed doses of pelvic radiotherapy, and to determine the safety of UFT plus oral leucovorin administration after pelvic radiotherapy, chemotherapy, and surgery. Standard treatment at M. D. Anderson Cancer Center for patients with T3, T4, and/ or > N1 rectal carcinoma is a preoperative continuous-infusion of fluorouracil (5 FU) with radiation therapy followed by four courses of 5-FU/ leucovorin postoperatively. Data suggest that UFT and leucovorin may offer a well-tolerated, fully oral treatment option that could be more convenient for patients. The trial presented herein provides data relative to the feasibility of preoperative oral UFT and leucovorin chemotherapy given during radiation therapy, and oral UFT and leucovorin chemotherapy following surgery in the treatment of patients with rectal cancer. This study is anticipated to serve as a pilot to develop an investigational treatment arm for a randomized trial of preoperative treatment of patients with rectal cancer. PMID- 9348572 TI - Current and future directions in adjuvant combined-modality therapy of rectal cancer. AB - Standard adjuvant therapy for transmural (T3) and/or node-positive rectal cancer is pelvic radiation therapy plus fluorouracil (5-FU)-based chemotherapy. Randomized trials are in progress to help determine the ideal chemotherapeutic agents and their optimal routes of administration in this setting, as well as to compare the efficacy and functional results of the pre- and postoperative bolus 5 FU/leucovorin combined-modality therapy approaches. New phase I trials will determine the recommended doses of tegafur and uracil (UFT) with oral leucovorin plus pre- or postoperative radiation therapy. PMID- 9348573 TI - Metastatic breast cancer: treatment with fluorouracil-based combinations. AB - During the 1990s, one in nine women in the western world will be diagnosed with breast cancer, and more than 58,000 will die of the disease each year in Europe alone. Recent changes in the primary therapy of operable breast cancer have not altered patient prognosis. Adjuvant therapy delays systemic recurrence and improves survival for only a fairly selected fraction of these patients. Therapy for metastatic breast cancer has not improved significantly in recent years. While combination chemotherapy may prolong survival in selected patients, few if any achieve cure. Standard chemotherapy regimens used to treat metastatic breast cancer, such as CMF (cyclophosphamide/methotrexate/fluorouracil), FAC (5 FU/Adriamycin/cyclophosphamide), and FEC (5-FU/epirubicin/cyclophosphamide), were developed over a decade ago. Current efforts to improve therapeutic efficacy have concentrated on decreasing drug toxicity and increasing drug doses (e.g., high dose chemotherapy with peripheral stem cell support). An important alternative to increasing therapeutic efficacy by such approaches is altering the administration schedules of well-known chemotherapeutic agents and introducing active new cytotoxic agents. One of the most frequently used cytotoxic drugs, 5-FU has documented activity in a variety of malignancies, most notably in breast cancer and gastrointestinal tract cancers. However, despite broad clinical experience, our knowledge of mechanisms of resistance in relation to various 5-FU schedules is limited. In vitro data and clinical experience show that resistance to one schedule of 5-FU can be overcome by using an alternative schedule, most often a protracted infusion. PMID- 9348574 TI - UFT in combination as adjuvant therapy for breast cancer. Grupo Oncologico de Sevilla Seville, Spain. AB - Between 1989 and 1993, 409 evaluable patients with breast cancer have been treated with tegafur and uracil (UFT) in an adjuvant setting in two different trials. Data from both trials were reviewed in December 1995 after a mean follow up of 5.09 +/- 1.1 years (range, 2.9 to 7.1 years). The aim of the first trial was to demonstrate the activity of UFT 400 mg/day for 6 months plus prednimustine 60 mg/m2 for 7 consecutive days, every 28 days in 6 cycles given orally (arm B). This scheme was compared with 6 cycles of cyclophosphamide 600 mg/m2, plus methotrexate 40 mg/m2, plus fluorouracil 600 mg/m2, every 4 weeks (arm A). In this study, 187 premenopausal women were evaluable, 96 in arm A and 91 in arm B, all of whom had positive axillary nodes. Although there were more younger patients in arm A than in arm B, prognostic factors were similar in both groups. Disease-free survival and overall survival were similar in both arms. However, some concern is raised by the low disease-free survival rate. The toxicity was mild (mainly nausea and vomiting and alopecia) and slightly worse in arm A. We believe that oral administration could be a useful alternative to the parenteral route. In the second trial, 222 evaluable patients received 20 mg/day of tamoxifen (Nolvadex) for 1 year (arm A), or the same dose of tamoxifen plus UFT 400 mg/day for 6 months. All patients were postmenopausal, and the characteristics of the tumors were the same as those in patients in the first trial. In arm A there were 109 patients and in arm B, 113. The groups were well balanced. The overall survival and the disease-free survival rates were equal in both arms, but were longer in arm B in the subset of patients with five or more axillary-involved nodes. The toxicity was negligible in both arms. We conclude that UFT/tamoxifen might be useful in postmenopausal patients with five or more involved nodes who are unable to follow a more aggressive schedule because of their age or low performance status. PMID- 9348576 TI - Future directions in the treatment of squamous cell carcinoma of the head and neck: the role of UFT. AB - Squamous cell carcinoma of the head and neck is a potentially curable neoplasm. Historically, the standard approach to treatment has been either surgery or radiation therapy, or a combination of the two. Over the past two decades, the role of chemotherapy in the curative approach to head and neck cancer has been evolving. Drugs like fluorouracil (5-FU), cisplatin (Platinol), methotrexate, paclitaxel (Taxol), docetaxel (Taxotere), and bleomycin (Blenoxane) have been investigated in both the palliative and curative settings. Multiple combination regimens have been used as neoadjuvant or induction regimens, but to date the original combination of cisplatin and fluorouracil is still the standard of therapy. UFT, a combination of uracil and tegafur (a fluorinated pyridine analogue), has been available in Japan as an oral form of treatment for squamous cell head and neck cancers. This article describes the rationale behind the use of UFT as an agent for the treatment of squamous cell carcinoma of the head and neck as well as data from ongoing investigations exploring its use in this disease. PMID- 9348575 TI - Preliminary results. UFT/methotrexate/leucovorin for breast Ca patients in progression after HDCT/PBPC support. AB - Twenty-four patients with metastatic breast cancer that had progressed after high dose chemotherapy with peripheral blood progenitor cell (PBPC) support were given intramuscular methotrexate in combination with oral UFT (tegafur and uracil) and oral leucovorin (the MUL regimen). Of the total treated, 21 patients are currently evaluable for response and toxicity. All patients had received extensive prior chemotherapy, including a high-dose regimen with PBPC support. Of the 21 assessable patients, 8 obtained either a complete response (1) or partial response (7), for an overall objective response rate of 38%. Another 7 patients had stable disease for 3 or more months. Therefore, the MUL regimen was able to stop disease progression for 3 or more months in nearly 75% of patients. The median time to progression and median overall survival from the start of MUL were 6 and 9 months, respectively. The toxicity was mainly gastrointestinal; 6 patients (29%) had World Health Organization grade 2/3 diarrhea, leading to a UFT dose reduction. Emesis was mild and easily manageable with thiethylperazine. In conclusion, MUL chemotherapy is active and well tolerated in patients with metastatic breast cancer in progression after high-dose chemotherapy. Further studies with this regimen, either as salvage chemotherapy or as maintenance chemotherapy after high-dose chemotherapy with PBPC, are warranted. PMID- 9348577 TI - Neoadjuvant therapy with cisplatin/fluorouracil vs cisplatin/UFT in locally advanced squamous cell head and neck cancer. AB - This study compared the activity and toxicity of fluorouracil (5-FU)/ cisplatin with the combination tegafur and uracil (UFT)/cisplatin in the neoadjuvant treatment of locally advanced-stage III or IV (MO)-head and neck cancer. A total of 67 patients were randomly assigned to treatment with cisplatin 100 mg/m2 on day 1 followed by either a continuous infusion of 5-FU 1,000 mg/m2/day on days 2 through 6 (group 1) or oral administration of UFT 300 mg/m2/day on days 2 through 20 (group 2). Both treatments were repeated every 21 days for four cycles. Responding patients received locoregional standard radiotherapy (50 to 70 Gy) after chemotherapy. Group 1 was comprised of 34 patients, 30 of whom were men, with a median age of 57.5 years; 79% of this group had a Karnofsky performance status of 90% to 100%; 70% had a squamous and 29% an undifferentiated histology. The majority (85%) had stage IV disease. Of the 33 patients in group 2, 29 were men. The median age was 56 years. Most (88%) had a performance status of 90% to 100%. More patients had a squamous than an undifferentiated histology (82% vs 18%) and most (88%) had stage IV disease. Overall response in group 1 was 73% (21% complete) compared with 79% (18% complete) in group 2. At a median follow-up of 84 months, no significant differences have emerged in overall survival, 15 vs 37 months, or time to progression, 8.5 vs 14.5 months, for groups 1 and 2, respectively. Toxicity was also similar, except for phlebitis, which occurred significantly more often in group 1 (71% vs 9%). Cisplatin/UFT was as effective as the classic cisplatin/5-FU regimen and has the advantages of outpatient oral administration and a lower incidence of phlebitis. PMID- 9348578 TI - Postoperative adjuvant chemotherapy for non-small-cell lung cancer. West Japan Study Group for Lung Cancer Surgery. The Japan Lung Cancer Research Group on Postsurgical Adjuvant Chemotherapy. AB - Given that no therapeutic methods of postoperative adjuvant chemotherapy for non small-cell lung cancer have been established, we selected UFT (tegafur and uracil) for investigation because UFT is less injurious to the host than intensive chemotherapies. The second study of the West Japan Study Group for Lung Cancer Surgery showed that the 5-year survival rates were 64.1% in the UFT group (UFT 400 mg/day for 1 year after surgery) and 49.1% in the control group (surgery alone). Thus, the survival rate for all patients was improved significantly in the UFT group as compared with the control group (log rank test, P = .033; generalized Wilcoxon test, P = .019). To establish the usefulness of long-term oral administration of UFT as an adjuvant therapy for completely resected non small-cell lung cancer, the nationwide Japan Lung Cancer Research Group on Postsurgical Adjuvant Chemotherapy is now conducting a comparative study of surgery alone versus surgery and UFT in patients with pathologic stage I adenocarcinoma. PMID- 9348579 TI - UFT plus cisplatin in advanced non-small-cell lung cancer: interim analysis of 67 patients. AB - A single-institution phase II study indicated that combination chemotherapy using UFT (tegafur and uracil) plus cisplatin (Platinol) in patients with non-small cell lung cancer was active with less host toxicity than other cisplatin-based therapies. To confirm these observations, the Japan JFT Lung Cancer Study Group conducted a multi-institutional phase II trial. The number of patients planned for this trial is 110. Eligibility includes previously untreated stage IIIB or IV non-small-cell lung cancer and a good performance status. UFT 400 mg/m2 in two divided doses is administered orally on days 1 through 14, and cisplatin 80 mg/m2 is injected IV on day 8. This treatment is repeated every 3 or 4 weeks. Between April 1995 and May 1996, 67 patients were enrolled, and all 67 were considered eligible for an interim analysis performed in October 1996. Among 63 patients evaluable for response, there was an overall response rate of 30% (95% confidence interval, 19% to 41%), with one complete response and 18 partial responses. With a median follow-up duration of 44 weeks, the median survival time was 32 weeks and the 1-year survival rate was 25%. Grade 3 leukopenia occurred in only 1 of 67 patients (1.5%), and there was no thrombocytopenia of grade 3 or greater. Vomiting, the most common nonhematologic toxicity observed, reached grade 3 or 4 in only 6 patients (9%). This interim analysis seems to support the observations of the previous single-institution phase II trial. PMID- 9348580 TI - UFT plus cisplatin for advanced gastric cancer. AB - In a laboratory study using an experimental peritoneum model of gastrointestinal cancer, the UFTP [tegafur and uracil (UFT) plus cisplatin] regimen was shown to provide a survival benefit compared with the UFTM (UFT plus mitomycin) regimen that has been considered standard chemotherapy for treatment of advanced carcinoma in Japan. Results indicated that a clinical benefit of the UFTP regimen could be expected. In the pilot clinical study, the UFTP regimen exhibited excellent antitumor effects and seems to improve survival. At present, we are conducting a phase II study in which the UFTP administration schedule was modified to reduce toxicity and secure a similar level of benefit compared with results of the pilot study. Positive results are anticipated. PMID- 9348581 TI - The UFT/leucovorin/etoposide regimen for the treatment of advanced gastric cancer. Oncopaz Cooperative Group. AB - Gastric cancer is the most chemosensitive adenocarcinoma among digestive neoplasms. A few years ago, we performed a phase II trial with the FLEP regimen, in which fluorouracil (5-FU) and leucovorin are combined with etoposide and cisplatin (Platinol). This regimen resulted in a 39% response rate and high toxicity. Then we used the combination UFT (tegafur and uracil)/leucovorin/etoposide: UFT 390 mg/m2/day orally on days 1 to 14; leucovorin 500 mg/m2 i.v. day 1, and 15 mg/12 h orally on days 2 to 14; and etoposide 100 mg/m2 i.v. on day 1 and then 200 mg/m2/day orally on days 2 and 3. Forty-six patients received a median of five courses. Five patients (11%) achieved a complete response and 12 (26%) a partial response, for an overall response rate of 37%. The response rate was 50% in patients with an Eastern Cooperative Oncology Group performance status of 0 to 1. Grades 3 to 4 toxicities appeared as follow: 17% of patients had diarrhea, 11% had nausea/vomiting, and 13% of patients had anemia. One patient died of neutropenia and sepsis. The median survival time was 9 months. In summary, UFT/leucovorin/etoposide is effective and moderately toxic in patients with advanced gastric cancer. A new trial with UFT/leucovorin/epirubicin is ongoing. PMID- 9348582 TI - UFT in gastric cancer: current status and future developments. AB - Despite recent progress in surgery and chemotherapy, advanced gastric cancer carries a poor prognosis. Although several antitumor agents have some clinical activity, responses are usually of short duration and fail to improve survival. Combination chemotherapy regimens containing fluorouracil (5-FU) and cisplatin (Platinol) frequently result in higher response rates, but fail to significantly alter the ultimate course of the disease. Tegafur and uracil (UFT) have been extensively studied in gastric cancer in Japan. In responding patients, single agent therapy results in a 1-year survival of 47%. Studies using combination regimens with UFT are currently performed in Europe, and data from Japan demonstrate that UFT can be safely combined with a variety of other agents. However, the exact contribution of UFT in these combinations will need to be evaluated further. The present review summarizes the use of UFT alone or in combination, as well as in the neoadjuvant and adjuvant settings, in the treatment of patients with advanced gastric cancer. PMID- 9348583 TI - A phase II trial. Oral UFT and leucovorin in patients with advanced gastric carcinoma. AB - Thirty-nine patients with locally advanced or metastatic gastric carcinoma received oral UFT (tegafur and uracil) plus leucovorin. Treatment consisted of UFT 360 mg/m2/day plus leucovorin 25 mg/m2/day, given orally in divided daily doses for 21 days followed by a 7-day rest period. The median age of the patients was 64 years, and the median World Health Organization performance status was 2. Patients received a median of two courses of treatment (range, 1 to 25). Among 37 evaluable patients, two patients achieved a complete response, and eight had partial responses, for an overall response rate of 27% (95% confidence interval, 15.4% to 42.9%). Stable disease was reported in 12 patients (32%) and another 15 showed disease progression. The median duration of response was 30 weeks, and the median duration of survival was also 30 weeks (range, 8 to 111). All patients were evaluable for toxicity. Significant toxicity (World Health Organization grade 3 or 4) included diarrhea in seven patients (18%), oral mucositis in six (15%), and nausea/vomiting in six patients. We conclude that oral UFT plus leucovorin, an outpatient regimen, has favorable activity in patients with gastric carcinoma and has tolerable toxicities. PMID- 9348584 TI - UFT plus leucovorin in advanced hepatobiliary tumors and pancreatic adenocarcinomas. AB - UFT (tegafur and uracil) has been studied extensively in Japan, with documented efficacy in hepatobiliary and pancreatic cancer. In the United States, UFT with or without leucovorin has not undergone phase II testing in these malignancies. Our current trial is designed primarily to assess the efficacy in terms of response rates to UFT with leucovorin in patients with advanced hepatobiliary and pancreatic cancer. Secondary objectives include determining response duration, time to disease progression, overall survival, quality of life, and toxicity. PMID- 9348585 TI - A pharmacoeconomic comparison of UFT and 5-FU chemotherapy for colorectal cancer in South America. AB - The escalating role played by managed care organizations in the health-care system is reflected in the increased demand for cost-effectiveness analyses (CEAs) to assess the balance between economic impact and clinical efficacy. For example, the high incidence and costs associated with colorectal cancer in Latin America calls for a comprehensive economic evaluation to ensure appropriate allocation of limited health-care funds. In addition, the current call for a "societal" perspective of such analyses indicates the need for increased consideration of the concerns of both patient and health-care provider. The introduction of oral tegafur and uracil (UFT) provided the opportunity to evaluate the pharmacoeconomic advantage of the new agent compared with the standard fluorouracil (5-FU). Results of this study indicated an economic advantage for oral UFT vs a 5-FU-based regimen in the treatment of colorectal cancer in Brazil and Argentina. It was further noted that the mild toxicity profile of UFT reduced both the number of clinic visits and the need for venipuncture procedures. Noting that oral UFT may have a positive impact on quality of life in addition to its estimated economic benefit, it was concluded that prospective economic research and quality-of-life evaluations are needed to fully assess the pharmacoeconomic impact of oral UFT. PMID- 9348586 TI - A multidisciplinary approach leads to system improvement. PMID- 9348587 TI - Computerized system streamlines chemotherapy order process. PMID- 9348588 TI - Computers link clinical trials registration and patient information. PMID- 9348589 TI - Medical identification cards facilitate emergency care for people with sickle cell disease. PMID- 9348590 TI - Nursing clinical research protocol education program. PMID- 9348591 TI - How healthcare professionals contribute to hope in patients with cancer. AB - PURPOSE/OBJECTIVES: To explore whether healthcare professionals influence the level of hope in patients with cancer and, if so, how they influence their hope. DESIGN: Descriptive, qualitative design. SETTING: An adult hematology/oncology unit in the upper midwestern United States. SAMPLE: Thirty-two men and women receiving active or supportive treatment or palliative care for cancer. METHODS: Semistructured interviews conducted in the participants' hospital rooms. Ten investigators and two consultants transcribed and analyzed the interview data using content analysis. They identified themes and subthemes that described healthcare professionals' roles. MAIN RESEARCH VARIABLES: Healthcare professionals' contributions to hope as described by patients with cancer. FINDINGS: Healthcare professionals positively and negatively influenced hope in this sample. Hope was facilitated by being present, giving information, and demonstrating caring behaviors. Negative influences on hope primarily concerned the way in which healthcare professionals gave information. CONCLUSION: Healthcare professionals do influence patients' perceptions of their hope. Although most nursing actions are hope enhancing, nurses can reduce a patient's sense of hope if information provided or attitude toward the patient is insensitive or disrespectful. IMPLICATIONS FOR NURSING PRACTICE: Nurses can increase patients' hope by being present, taking time to talk, and being helpful. They must provide information and answer questions in a compassionate, positive, honest, and respectful manner. Caring behaviors such as thoughtful gestures, showing warmth and genuineness, and being friendly and polite also increase patients' hope. PMID- 9348592 TI - Current status of hormonal treatments for metastatic breast cancer in postmenopausal women. AB - PURPOSE/OBJECTIVES: To review hormonal treatments for advanced breast cancer in postmenopausal women. DATA SOURCES: Published manuscripts, oncology journals, and clinical experience. DATA SYNTHESIS: Traditionally, hormonal treatments have included surgery and manipulation of estrogen, progestin, or androgen levels. Selective aromatase inhibitors have the advantage of a low side effect profile and offer a new treatment option for advanced breast cancer. CONCLUSIONS: New drug development has made a significant contribution to safe and effective oral treatments for advanced breast cancer with improved side effect profiles. IMPLICATIONS FOR NURSING PRACTICE: An increased awareness of hormonal treatments in advanced breast cancer will help nurses educate, counsel, and promote patient awareness and acceptance. PMID- 9348594 TI - Reciprocal support in the context of cancer: perspectives of the patient and spouse. AB - PURPOSE/OBJECTIVES: To examine the relationship between mutual spousal support and psychological health status of patients and their spouses. DESIGN: Cross sectional, descriptive. SETTING: Outpatient oncology clinics (physician, hospital, state cancer center) in the southwestern United States. SAMPLE: A convenience sample of 73 predominantly Caucasian couples with one dyad member receiving treatment for cancer. The mean age of the husbands and wives was 60.7 years and 57.1 years, respectively. METHODS: Questionnaires (Interpersonal Relationships Inventory; Disease Course Graphic Scale; Rosenberg Self-Esteem Scale; Centre for Epidemiological Studies-Depression Instrument; demographic form) completed by patients and spouses. MAIN RESEARCH VARIABLES: Marital reciprocal support, interpersonal support, severity of illness, interpersonal conflict, self-esteem, and depression as perceived by the patients and spouses; performance status as assessed by the patients. FINDINGS: The patients perceived more interpersonal support than the spouses. Both experienced more self-esteem when marital reciprocal support was balanced and high rather than unbalanced or balanced but low. A similar pattern was found for the spouses for self-esteem and depression regarding interpersonal support. Marital and interpersonal support decreased and depression increased as interpersonal conflict increased. CONCLUSIONS: Mutual spouse support is significantly related to patient and spouse self-esteem and depression. Marital reciprocal support is negatively related to interpersonal conflict. More attention must be given to family-oriented oncology nursing care. IMPLICATIONS FOR NURSING PRACTICE: Include the patients' and spouses perceptions of balance in the exchange of marital support in family assessments. Intervene directly or through referral to enable couples to understand the benefit and obligation to support each other when faced with a life-threatening illness. PMID- 9348593 TI - Decision making by parents and healthcare professionals when considering continued care for pediatric patients with cancer. AB - PURPOSE/OBJECTIVES: To better define the treatment-related decisions considered most difficult by parents of pediatric patients with cancer and the factors that influenced their final decisions. DESIGN: Retrospective-descriptive design. SETTING: Pediatric oncology institution in the mid-southern region on the United States. SAMPLE: 39 parents representing 37 of 83 eligible families, 16 attending physicians, three nurses, and two chaplains. METHODS: Parent participants responded by telephone to six open-ended interview questions and a 15-item questionnaire about factors that were important when making the decision to continue care. Healthcare professionals were interviewed face-to-face. MAIN RESEARCH VARIABLES: Most difficult treatment-related decisions; factors influencing decision making. FINDINGS: Parents reported 15 types of difficult decisions, the majority of which were made late in the course of treatment. Deciding between a phase I drug study or no further treatment (n = 14), maintaining or withdrawing life support (n = 11), and giving more chemotherapy or giving no further treatment (n = 8) were the most frequently reported difficult decisions. Parents rated "recommendations received from healthcare professionals" as the questionnaire factor most important in their decision making, and healthcare professionals rated "discussion with the family of the patient" as the most important factor. CONCLUSION: Parents of children or adolescents with cancer and their healthcare providers face difficult treatment-related decisions, many of which occur late in the course of treatment. Parents and healthcare professionals cite similar factors in their decision making but differ in their ratings of the factors' importance. For parents, the information and recommendations they receive from healthcare professionals figure most frequently and most importantly in their decision making. For healthcare professionals, the certainty that the patient will not get better and discussions with the patient's family figure most importantly in their decision making. Once parents conclude that their child can not get better, they are more likely to choose noncurative options such as choosing no further treatment or withdrawing life support. IMPLICATIONS FOR NURSING PRACTICE: Nurses can help determine what information parents need in their decision making. Particular attention must be given to ways to communicate the likelihood of the their child's survival. PMID- 9348595 TI - Controlling conditioning-related emesis in children undergoing bone marrow transplantation. AB - PURPOSE/OBJECTIVES: To compare the efficacy of two antiemetic regimens, ondansetron alone versus perphenazine with diphenhydramine, on emesis control in children undergoing conditioning therapy for bone marrow transplantation (BMT). DESIGN: Single-center, prospective, open, randomized, crossover study. SETTING: Pediatric BMT unit in an urban area in the northeastern United States. SAMPLE: 28 children, ages 4-17, undergoing BMT for a variety of underlying diseases. METHODS: After randomization to one of the two antiemetic regimens, emesis control was evaluated during conditioning therapy. If a participant experienced more than five episodes of emesis during any 12-hour period, he or she was crossed over to the other antiemetic regimen. If emesis control still was not achieved, the participant was removed from the study and other medications were administered to control vomiting. MAIN RESEARCH VARIABLES: Number of emetic episodes and incidence of side effects. FINDINGS: 10 of 15 patients (67%) who received ondansetron experienced major emesis control (no more than two episodes) compared with 0 of 13 patients (0%) who received perphenazine with diphenhydramine (p = 0.044, Fisher exact test). Of those who crossed over to ondansetron after failure with perphenazine and diphenhydramine, 38% were able to achieve major emesis control. CONCLUSIONS: Ondansetron offers superior antiemetic control over the combination of perphenazine and diphenhydramine for children undergoing high-dose chemotherapy with or without total body irradiation for BMT. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses must develop an understanding of the etiology of therapy-induced emesis and the mechanisms of action of the various classes of antiemetic agents designed to control it. Implementing documentation to describe events of emesis will help to tailor antiemetic therapy to a patient's specific situation. Further research is necessary to determine alternate strategies, including different combinations or sequences of antiemetics to provide optimum emetic control during acute and delayed phases of emesis. The higher cost of ondansetron therapy must be considered within the context of superior efficacy. PMID- 9348596 TI - Pattern of association over time of side-effects burden, self-help, and self-care in women with breast cancer. AB - PURPOSE/OBJECTIVES: To describe the side-effects burden experienced over time by 53 women who were receiving treatment for breast cancer, and to describe the association of side-effects burden with self-help and self-care. DESIGN: Data were drawn from the Self-Help Intervention Project (SHIP), an intervention study designed to test the effectiveness of nursing interventions for women who were receiving treatment for breast cancer. SETTING: Subjects were interviewed in their homes or treatment locations three times over a period of four to five months. SAMPLE: 53 women randomly assigned to the SHIP control group. METHODS: The researchers collected data at a designated period of time after treatment was initiated (Time 1), six to eight weeks later (Time 2), and three months after that (Time 3). MAIN RESEARCH VARIABLES: Side-effects burden, as measured by items from the Symptom Transition Scale and the Side Effects Checklist; self-help, as measured by the Inventory of Adult Role Behavior; and self-care, as measured by the Inventory of Adult Self-Care Behaviors and the Self-Care Inventory-Wellness Promotion subscale. FINDINGS: Fatigue was the most frequent and problematic side effect over time. Other frequent and problematic side effects over time included sore arm(s), difficulty sleeping, and skin irritation. Significant correlations were evident for self-help with symptom extension, number of side effects, depression, difficulty concentrating, and pain. No significant relationships were evident between self-care and an increase in side effects. Small relationships existed for self-care between symptom extension at Time 2 and Time 3. CONCLUSIONS: Over time, side effects interfered with patients' ability to perform adult role activities. For the most part, problematic side-effects burden was not associated with self-care at any point in time. The scattered associations that did exist were in the negative direction. IMPLICATIONS FOR NURSING PRACTICE: A need exists for clinically individualized nursing interventions to reduce the side-effects burden of women receiving treatment for breast cancer. Interventions can do much to reduce the perception of illness severity so that self-help and self-care can be maintained. PMID- 9348597 TI - Bone marrow transplant nurses' knowledge, beliefs, and attitudes regarding pain management. AB - PURPOSE/OBJECTIVES: To measure bone marrow transplant (BMT) nurses' knowledge, beliefs, and attitudes regarding pain management. DESIGN: Descriptive, exploratory. SETTING: A 32-bed BMT unit in a 567-bed tertiary-care institution located in the midwestern United States. SAMPLE: 39 BMT nurses (20 pediatric, 19 adult). The mean length of BMT experience was 7.05 years. METHOD: BMT nurses completed a 49-item, investigator-developed questionnaire. MAIN RESEARCH VARIABLES: Nurses' knowledge, beliefs, and attitudes related to pain management. FINDINGS: Many BMT nurses had high knowledge levels and positive beliefs and attitudes related to pain management. The mean of correct responses to knowledge items was 79%. Nurses had a high knowledge level of pain assessment, but only 74% indicated that patient self-report of pain is the most reliable indicator of pain. The majority of the nurses' responses were congruent with literature sources regarding the onset of mucositis pain, self-report of pain, and opioid tapering. Most nurses agreed that pain management is rewarding and satisfying; fewer agreed that it is not stressful. Nurses' requests for information focused on opioid therapy. CONCLUSIONS: The investigators identified specific knowledge gaps. The variability of scores indicated that some nurses are more expert than others regarding pain management and therefore could be resources for other nurses. IMPLICATIONS FOR NURSING PRACTICE: Educational offerings can increase knowledge and promote positive beliefs and attitudes among BMT nurses, thereby enhancing pain management. PMID- 9348598 TI - Breast cancer treatment-related patterns in side effects, psychological distress, and perceived health status. AB - PURPOSE/OBJECTIVES: To describe variations in side effects of treatment of breast cancer and concurrent variations in psychological distress and perceived health status. DESIGN: Data were collected at six collection points: 7-10 days, one month, two months, three months, six months, and one year postsurgery. Standardized inventories, including subscales from the Psychosocial Adjustment to illness Scale, the Multilevel Assessment Inventory, and the investigator developed Treatment Recovery Inventory, were completed by subjects in their homes at the six data-collection points. SETTING: Self-report measures completed in subjects' homes. Private practices of breast surgeons at medical centers in the New York City metropolitan area SAMPLE: Convenience sample of 93 women from a main longitudinal study (x = 51.4 years of age). MAIN RESEARCH VARIABLES: Psychological distress, perceived health status, and side effects of treatment for breast cancer. FINDINGS: Fatigue and emotional distress were persistent issues. Psychological distress and perceived health status improved with no significant differences between breast-conserving and mastectomy groups. Significant differences existed between patients receiving chemotherapy versus no therapy between three and six months. Significant changes also existed in perceived health status and psychological distress in both the positive and negative node groups with no differences between groups. CONCLUSIONS: Adjustment, a multidimensional and complex process, occurs over time. IMPLICATIONS FOR NURSING PRACTICE: Patterns in side effects should enable clinicians to anticipate needs for planning methods of control over time. Nurses must assess side effects from a multidimensional perspective at all phases. PMID- 9348599 TI - Concerns of rural men and women experiencing cancer. AB - PURPOSE/OBJECTIVES: To examine the interpersonal relationships, self-image, healthcare interactions, and occupational concerns of rural men and women experiencing cancer directly or as a caregiver of a person with cancer. DESIGN: A descriptive design employing a mail survey as part of a larger, longitudinal study. SETTING: Rural areas in the northern Rocky Mountain region of the United States. SAMPLE: 294 people with cancer and 294 family caregivers in Montana responded to mail questionnaires. Fifty-two percent were women, and almost all were Caucasian. METHODS: Investigator-developed Cancer Concerns Inventory mailed to participants. MAIN RESEARCH VARIABLES: Interpersonal relationships, self image, healthcare interactions, occupational concerns, gender, and type of experience with cancer. FINDINGS: In general, women were more likely than men to report relationship problems, lack of support, and feelings that were not understood. People with cancer, as compared with caregivers, were more likely to report feeling alone and that other people avoided them and were afraid to talk to them. Men with cancer were more likely than women with cancer and caregivers to feel that their job security was threatened. However, only a small percentage of all participants felt discriminated against at work. In general, a higher percentage of women with cancer and men caregivers reported concerns about healthcare interactions than men with cancer and women caregivers. CONCLUSIONS: Men and women caregivers and people with cancer have different concerns regarding cancer. IMPLICATIONS FOR NURSING PRACTICE: Nurses working in rural areas must help families work through relationship difficulties, maximize healthcare interactions, and be an advocate for people with cancer. PMID- 9348600 TI - Bacterial resistance to antibiotics: it's our problem. PMID- 9348601 TI - Physicians' views on pediatric preventive dental care. AB - Physicians who provide primary care for children are considered to be in a unique position to provide dental preventive care to their patients. No literature relates the amount of preventive oral health education that physicians receive during training. The purpose of this study was to assess the knowledge, attitudes, and beliefs of pediatricians and family physicians toward preventive dental care in children. A questionnaire was mailed to 398 pediatricians and 632 family physicians licensed to practice in the state of Alabama. The response rate after one mailing and a reminder was 46%. Physician's knowledge about many aspects of preventive dental care was good, but areas of great concern were identified. Overall, most respondents received 2 hr or less of preventive dental education during medical and specialty training. Pediatricians were better informed than family physicians in the areas of general dental knowledge and prevention counseling related to oral health (P < 0.05). PMID- 9348603 TI - Treatment of Class III problems begins with differential diagnosis of anterior crossbites. AB - Etiology of Class III malocclusion can be genetic or environmental. Proclination of mandibular incisors and retroclination of maxillary incisors can cause posturing of the mandible in an anterior position due to incisal interference, a condition called pseudo Class III malocclusion that can be misleading in evaluating a patient with skeletal Class III malocclusion. Unfortunately, cephalometric evaluation may not be the most reliable tool in differentiating whether the maxilla or the mandible contributes to the skeletal disharmony. The most consistent findings seem to be the dental characteristics of Angle's Class III molars and canines, retroclined mandibular incisors, and the presence of an edge-to-edge or an anterior crossbite occlusion. This paper presents a diagnostic scheme to differentiate between dental and skeletal crossbites. Early treatment of Class III malocclusion can help to minimize the adaptations and limitations that are often seen in severe malocclusion of the late adolescence. However, treatment of skeletal crossbites remains a continuous challenge to the profession. Due to the diversity and variability in facial growth, accurate individualized growth prediction is not possible at the moment. Treatment directed at the mandible seems to invite relapse during the pubertal growth period. Treatment directed at the maxilla shows promising results and is awaiting long-term clinical results following early orthopedic interventions. Several intraoral appliances have proved to be successful in eliminating dental crossbites. PMID- 9348602 TI - Implications of evidence-based practice on preventive procedures in pediatric dentistry. PMID- 9348604 TI - Clinical and microbial considerations for the treatment of an extended kindred with seven cases of prepubertal periodontitis: a 2-year follow-up. PMID- 9348605 TI - Acute ethanol toxicity from ingesting mouthwash in children younger than 6-years of age. AB - The purpose of our study was to analyze reports of the American Association of Poison Control Centers (AAPCC) of suspected overingestion of ethanol from mouthrinses by children younger than 6 years of age between 1989 and 1994. Annual incidence rates of reported ethanol exposures attributed to mouthrinses were calculated. Lethal and toxic amounts of several mouthrinses were calculated using peak blood ethanol concentrations of 500 and 50 mg per 100 mL, respectively. In 1994, there were 2937 calls reported by poison control centers related to ethanol containing mouthrinses, an estimated incidence of 168 reported exposures per 100,000 children younger than 6 years of age. A 15-kg child who ingests 212 mL (7.2 oz.) of Listerine (26.9% ethanol) ingests 57 mL (1.9 oz.) of ethanol, which is potentially lethal. Approximately one-tenth that amount of ethanol can produce a toxic reaction. Physicians, dentists, and other health care providers should inform parents of the dangers associated with accidental ingestion of mouthrinse and encourage them to keep mouthrinse out of the reach of children. The Food and Drug Administration (FDA) should require readily visible warning labels and child resistant caps for containers with potentially toxic volumes of ethanol. The American Dental Association (ADA) should re-evaluate its acceptance criteria for advertising cosmetic mouthrinses in its publications and consider including child resistant caps and warning labeling. PMID- 9348606 TI - Gastroesophageal reflux and dental erosion: case report. PMID- 9348607 TI - A review of selected microstomia prevention appliances. AB - Perioral burns may occur due to electrical, thermal, or chemical agents. The resultant contracture of the facial tissue during healing causes limited oral access, compromised esthetics, and other related problems. This article presents various microstomia prevention appliances used by dentists and hospital burn centers. These appliances reflect different treatment concerns, ease of fabrication, age appropriateness, and cost effectiveness. An understanding of these factors and available appliances will aid the clinician in selecting or developing the best appliance for burn patients. PMID- 9348608 TI - Surface hardness of a resin composite cured with a transparent cone. AB - The purpose of this study was to evaluate the surface hardness of a resin composite (TPH) polymerized with and without the use of a transparent light curing cone. Twenty composite blocks were made on a Teflon mold with cylindrical holes of 3 mm depth and 6 mm radius, and were light cured following different procedures. Group 1: Ten samples were pressed with a glass slide and light cured for two 40-sec exposures without the plastic cone; Group 2: Ten samples were light cured using the transparent plastic cone which was pressed down into the composite until the tip was 1 mm from the floor of the cylindrical Teflon mold. The curing light was activated for 40 sec. The cone was then removed and the remaining part of the mold was filled in one portion, pressed with a glass slide, and light cured for 40 sec. After curing, the samples were placed in distilled water for 48 hr. The hardness of the samples was then measured with a Rockwell Hardness Tester at three different points on each composite block; therefore, 30 measurements per group were taken. The data were statistically analyzed using an unpaired Student's t test. The results revealed that the resin composite cured with the transparent plastic cone had a statistically significant higher surface hardness value (P < 0.0001) than the group cured without the cone. PMID- 9348609 TI - An alternative restorative method for regional odontodysplasia: case report. AB - A 5-year-old Caucasian male presented with early loss of multiple deciduous teeth. All the characteristics were consistent with the diagnosis of regional odontodysplasia (ROD). Significant initial findings included premature loss of multiple primary mandibular teeth and some malformed permanent teeth. The affected teeth showed hypoplastic enamel and dentin, short roots, and wide pulp chambers, and were localized in the mandibular dentition. Treatment objectives for this patient were to provide improved esthetics, restored chewing function, and space maintenance by the construction of a temporary prosthetic restoration. However, with limited tooth support and an unusual occlusal pattern, it is difficult to obtain satisfactory retention and esthetics with traditional prosthetic techniques. In this article we introduce an alternative method for fabricating a custom removable denture and discuss the prognosis of the malformed permanent dentition and further treatment plan. PMID- 9348610 TI - Periodontal breakdown and pathologic root resorption of primary molars following traumatic injuries to the chin: case report. PMID- 9348611 TI - Temperament and trait anxiety as predictors of child behavior prior to general anesthesia for dental surgery. AB - Children's individual styles of interaction with the environment (temperament) influence stable tendencies towards distress (trait anxiety) and context-specific manifestations of distress (state anxiety). Measures of temperament and trait anxiety were examined as predictors of state anxiety (i.e., disruptive behaviors) in the presurgical setting. During a 2-month period, 51 nonpremedicated, healthy children (M = 3 years of age) were consecutively studied-as they presented to a hospital setting for dental treatment under general anesthesia (GA). Using correlation and backward multiple regression analyses, one temperament category (shyness), but not trait anxiety (the revised CMAS), predicted disruptive behaviors (the revised MBPRS) during preseparation (r2 = .16, P = .0038) and separation (r2 = .09, P = .0281) from parents. Shyness, age, and gender best predicted disruptive behaviors during preseparation (multiple R2 = .31, P = .0005). Temperament (a) predicts children's distress in the presurgical setting, and (b) appears to be moderated by age, gender, and interpersonal factors. Awareness of temperamental influences can help predict children's behavior and aid in the presurgical care of children. PMID- 9348612 TI - Conversion rate of research abstracts to publications in pediatric dentistry. PMID- 9348613 TI - Laparoscopic Nissen fundoplication at a teaching center: prospective analysis of 103 consecutive patients. AB - The Nissen fundoplication is the most extensively studied and successfully employed surgical solution to gastroesophageal reflux disease (GERD). Early success with the application of minimally invasive techniques to this procedure has been reported by several authors. One hundred three consecutive patients were operated on for the symptoms and complications of GERD. Preoperative evaluation consisted of esophagogastroduodenoscopy and esophageal manometry. Twenty-four hour esophageal pH was obtained selectively. All cases were performed in a traditional training environment, adhering to techniques previously described in the open literature. Clinical data consisted of operative time, postoperative hospital days, days to resumption of normal activities, and morbidity. Patients were followed clinically for the incidence of dysphagia, bloating, and recurrent reflux symptoms. These were graded using a modified Visick score prior to discharge, at 1, 3, and 6 months, and then annually. All patients underwent completion of their procedure; however, four required conversion to open technique and were excluded from analysis. Mean operative time for the 99 laparoscopic procedures was 180 min. Mean operative time was significantly longer for the first 50 cases (202.1 min) than the last 49 (164.2 min). Mean postoperative hospital stay was 2.3 days with 10 days to resumption of normal activities. Mean follow-up was 15 months (range 3-39 months). Three of the four treatment failures underwent open revision with good subsequent results. Patient satisfaction as reflected by the modified Visick score reveals 96% good to excellent results (Visick 1 or 2) with no persistent dysphagia. The Nissen fundoplication can be safely performed using minimally invasive techniques with the benefit of postoperative recovery typical of other laparoscopic procedures. By strictly adhering to the primary technical principles previously described in the open approach, early results are comparable. The procedure can be safely performed in a traditional training environment. The modified Visick system is a simple and effective method to quantify postoperative patient satisfaction. PMID- 9348614 TI - Laparoscopic cholecystectomy at cesarean section. A new surgical option. AB - A 29-year-old woman with recurrent cholelithiasis in pregnancy and a history of previous cesarean section underwent elective repeat abdominal delivery combined with laparoscopic cholecystectomy under general anesthetic. A transverse suprapubic incision was employed for fetal extraction and for facilitating the placement of three upper abdominal laparoscopic cannulas. After closure of the laparotomy incision, a pneumoperitoneum was established, and the gallbladder was removed laparoscopically. The surgery was uneventful, and the patient was discharged on the third postoperative day. PMID- 9348615 TI - Preoperative Nyhus classification of inguinal hernias and type-related individual hernia repair. A case for diagnostic laparoscopy. AB - The goal of this prospective study was to determine the clinical value (sensitivity and specificity) of preoperative hernia classification (Nyhus classification) using three distinct methods: clinical examination, Doppler ultrasonography, and diagnostic laparoscopy. Thirty patients with 35 suspected groin hernias were included. Definitive hernia classification was achieved by laparoscopic peritoneal incision and dissection of the inguinal floor. Twenty eight laparoscopic hernia repairs followed. Sensitivity and specificity were calculated for each preoperative evaluation method. Clinical examination was found to be more accurate than Doppler ultrasonography. The highest scores for sensitivity (0.93) and specificity (1.00) were achieved, however, by diagnostic laparoscopy. Therefore, the authors consider diagnostic laparoscopy to be a valuable preoperative tool for assessing hernia type. An accurate preoperative hernia classification will allow an individualized type-related hernia repair (open: anterior, posterior approach, or laparoscopic: transabdominal preperitoneal, total preperitoneal, inner-ring closure, mesh insertion). PMID- 9348616 TI - Can hypothermia be evidenced during laparoscopic cholecystectomy? AB - Maintenance of pneumoperitoneum to perform laparoscopic surgery with carbon dioxide (CO2) could induce hypothermia. The authors assessed the mean body temperature (MBT) and changes in total body heat content (TBHC) under laparoscopic cholecystectomy. Thirty-six ASA I-II female patients underwent open cholecystectomy (Group-O, n = 18) or laparoscopic cholecystectomy (Group-L, n = 18). Esophageal temperature and four skin-surface temperatures were measured before induction and then every 10 minutes, and at arrival to the postanesthesia care unit. Operating room temperature was 22.9 +/- 1.2 degrees C. Operating time was as follows: G-O, 74 +/- 21 minutes; G-L, 94 +/- 16 minutes. After 60 minutes of surgery, decrease in TBHC was as follows: G-L = 54.9 kJ and G-O = 40.9 kJ. Decrease in MBT after 60 minutes intervention and at arrival to the postanesthesia care unit was as follows: G-L = 0.13 and 0.66 degree C and G-O = 0.17 and 0.49 degree C, respectively. There were no statistically significant differences between groups. Minor differences were attributed to longer surgery duration in the laparoscopic group. PMID- 9348617 TI - Transinguinal laparoscopic examination of the contralateral groin in pediatric herniorrhaphy. AB - The authors laparoscopically assessed the contralateral groin (CG) via the symptomatic inguinal hernia in 91 patients to avoid unnecessary CG exploration and to allow for identification of asymptomatic CG hernias. Once the symptomatic hernia sac was opened, a 4.5-mm trocar was placed intraperitoneally and the CG internal ring was inspected with a 4-mm laparoscope. When compared with the authors' previous surgical policy for routine CG exploration in children younger than 2 years of age, laparoscopic findings altered the procedure performed in 42 of 91 patients (46.2%). In patients younger than 2 years of age, the CG did not require repair of an unsuspected hernia in 55.9% of patients based on laparoscopic findings. Conversely, in children 2 years of age or older, 40.4% required CG repairs of unsuspected hernias or patent processus vaginalis (PPV). Transinguinal laparoscopic examination in pediatric herniorrhaphy provides important information about the CG without the need for additional trocar sites. PMID- 9348618 TI - Adenomyoma of the ampulla of Vater: an uncommon cause of bile duct obstruction. AB - Adenomyomas of the bile ducts are extremely rare. They are most likely often overlooked also when situated in the ampulla of Vater and obstructing bile flow. Of 3,131 endoscopic retrograde cholangiograms, four patients with ampullary adenomyoma and signs of biliary obstruction have been diagnosed. When recognized they can be treated effectively by endoscopic means. However, their radiologic and endoscopic appearances were found difficult to interpret and led to unnecessary endoscopic sessions in one patient and surgery in another. Thus, increased awareness of this entity is important to avoid overlooking or misdiagnosing it. PMID- 9348619 TI - Laparoscopic Hill's vagotomy by the abdominal wall lifting method. AB - We performed laparoscopic Hill's vagotomy by the abdominal wall lifting method in nine patients with intractable duodenal ulcer. Our original I-type lifting bar is a curved stainless-steel rod 5 mm in diameter. One I-type lifting bar is inserted intraperitoneally into each of the right and left hypochondrial regions. An incision is made in the lesser omentum near the gastroesophageal junction, and the right esophageal wall and right crus of the diaphragm are dissected and exposed. The posterior trunk of the vagus nerve is identified and divided. Then the neurovascular bundle is dissected and divided repeatedly along the lesser curvature of the stomach from the first branch of the crow's foot to the gastroesophageal junction. The mean operating time was 163 min, with little blood loss. The reduction rate of basal acid output and maximal acid output was, respectively, 73.7 +/- 0.1 and 63.7 +/- 0.1%. Four weeks after surgery, gastroduodenoscopy revealed ulcer healing to a scar. PMID- 9348620 TI - An experimental model of cellular aerosolization during laparoscopic surgery. AB - Laparoscopic surgery for cancer has led to the unwelcome occurrence of malignant seeding of port sites. It is hypothesized that this seeding may be a result of aerosolization and forced egress of cells from the peritoneum as a result of pneumoperitoneum. The purpose of this study was to develop a model that would allow for future investigations of cellular aerosolization. Six swine were anesthetized, intubated, and ventilated. A port was placed in the midline and the abdomen insufflated. After insufflation a 14-gauge angiocath was placed in the abdomen through a separate site and attached to a closed system that allowed escaping air to bubble through 3 ml of saline. Intraabdominal pressure was serially increased at 30-min intervals to 8, 10, 12, 14, 16, and 18 mm Hg, and separate saline samples were collected at each interval. Saline samples were centrifuged, and epithelial cells were counted by direct vision and Giemsa staining. Epithelial cells were recovered at all levels of pneumoperitoneum. There was a moderate correlation between the level of pneumoperitoneum and the number of cells collected (r = 0.61, p < 0.19). Results of this study suggest that during pneumoperitoneum there is an ongoing egress of aerosolized cells from the abdomen. Application of this model may aid in future study of aerosolization of cancer cells during laparoscopic surgery. PMID- 9348621 TI - Cholecystoenteric fistula is not a contraindication for laparoscopic cholecystectomy: report of five cases treated by laparoscopic approach. AB - The authors present five cases (three female, two male, mean age 50.8) of cholecystoduodenal fistula incidentally discovered during laparoscopic cholecystectomy and treated by laparoscopic approach. The laparoscopic technique adopted is described and all patients recovered promptly with no immediate or long-term post-operative complications. Discharge from the hospital was after 4.5 days, and after 6 months follow-up all patients were in good clinical condition. These results indicate that when the surgeon is skilled in advanced laparoscopic operative techniques such as duodenal mobilization and intracorporeal suturing and knotting, cholecystoduodenal fistula can no longer be considered a contraindication for laparoscopic treatment. PMID- 9348622 TI - Laparoscopic versus open cholecystectomy in acute cholecystitis. AB - Elective laparoscopic cholecystectomy is established as the treatment of choice for symptomatic cholecystolithiasis and is now proposed for the treatment of acute cholecystitis. We initiated the present study in order to clarify the question of safety of the procedure in the presence of an inflamed gallbladder, and to compare the results with those of a traditionally treated group with acute cholecystitis. We compared the preoperative, operative, and postoperative courses of 146 patients with acute cholecystitis, managed laparoscopically between 1994 and 1996, with those of 97 patients, treated traditionally by open cholecystectomy for the same diagnosis between 1992 and 1993. In the acute cholecystitis cases, when laparoscopic cholecystectomy was successfully performed, the operative and postoperative courses were superior to those of open cholecystectomy. The use of drains and NG tubes, the need for antibiotics and analgesia, the associated morbidity, and the hospital stay were significantly reduced. Following conversion, the postoperative course was similar to that of open cholecystectomy. Of the group of acute cholecystitis cases laparoscopically approached 39 (27%) needed conversion. Twenty-five complications occurred in 24 (16.5%) patients of the laparoscopic group, whereas 30 complications occurred in 25 (26%) patients of the traditionally operated group. Male sex, older patients, and larger bile stones were found to be associated with a higher conversion rate as well as a higher complication rate. A nonpalpable gallbladder and gangrenous cholecystitis were associated with conversion while fever was associated with complications. Laparoscopic cholecystectomy can be performed safely in selected cases of acute cholecystitis, with acceptable conversion and low complication rates. When laparoscopic cholecystectomy is successfully performed, the operative and postoperative courses are superior to those of open cholecystectomy. Following conversion, the postoperative course is similar to that of open cholecystectomy. According to this study, male sex, older age, large bile stones, a nonpalpable gallbladder, and gangrenous cholecystitis may be regarded as predictors of conversion, while male sex, older age, large bile stones, and fever may be regarded as predictors of complications. The timing of laparoscopic cholecystectomy should be within 96 h from onset of the inflammation. PMID- 9348623 TI - Simultaneous hemodynamic and echocardiographic changes during abdominal gas insufflation. AB - The purpose of this study was to investigate cardiovascular changes during CO2 pneumoperitoneum. We performed simultaneous hemodynamic recordings and transesophageal echocardiographic measurements of possible alterations in cardiac dimensions. Seven patients scheduled for elective laparoscopic cholecystectomy were investigated. With an intraabdominal pressure of 15 mm Hg, mean arterial pressure increased from 75 to 93 mm Hg (p < 0.05). Despite the increase in pulmonary capillary wedge pressure (PCWP) from 10 (9.5-12) to 17 (16-19.9) mm Hg (p < 0.05), left ventricular end-diastolic area index (EDAI) did not change significantly. The cardiac index remained unchanged. Thus abdominal gas insufflation substantially alters the PCWP/EDAI relation. During pneumoperitoneum, left ventricular filling pressure, estimated by PCWP, cannot be used as an indicator of left ventricular dilation. PMID- 9348624 TI - Laparoscopic jejunostomy with an 18-mm trocar. AB - We describe our technique to perform laparoscopic jejunostomies with an 18-mm trocar. This procedure facilitates the exteriorization of the proximal bowel and construction of the jejunostomy. We describe our laparoscopic technique in nine patients with severe neurologic conditions (two in the postoperative period of a cerebral aneurysm in a coma, three patients with severe head injury, and four patients with cerebrovascular strokes). The operative time ranged from 20 to 75 min (average, 44.38 min). Nutrition was initiated 24 h after the placement of the jejunostomy. Tolerance of the enteral nutrition was excellent in all cases. One major complication occurred, minor leakage around the feeding tube 3 weeks after the jejunostomy was constructed. The jejunostomy was removed without further consequences. Laparoscopy is an effective technique for the creation of feeding jejunostomies. We believe that this minimally invasive approach is an alternative for patients requiring long-term postpyloric enteral feeding. PMID- 9348625 TI - Laparoscopy-assisted sigmoid colectomy for volvulus. AB - We report on five male patients with sigmoid volvulus treated by laparoscopy assisted sigmoid colectomy. Intraoperative colonic irrigation was used in two patients prior to resection and primary anastomosis. An intracorporeal technique of bowel anastomosis was used that allowed smaller skin incisions as compared with the extracorporeal technique. All five patients recovered uneventfully, and most were discharged within 1 week after the operation. There was no recurrence at the conclusion of the follow-up. Laparoscopy-assisted sigmoid colectomy may prove to be the procedure of choice in patients with sigmoid volvulus. PMID- 9348626 TI - Laparoscopic management of ruptured pyogenic liver abscess. AB - Intraperitoneal rupture of pyogenic liver abscess is a rare but potentially fatal disease, often involving the elderly, who are commonly of poor surgical risk with background of significant medical illness. Accurate preoperative diagnosis is difficult and often necessitates exploratory laparotomy for peritonitis. We report a case of ruptured cryptogenic solitary liver abscess presented with acute peritonitis that was successfully treated laparoscopically. The laparoscopic approach obviated the access trauma, confirmed the diagnosis, was able to drain the abscess, as well as provided microbiology and histology samples. A thorough peritoneal lavage could be also performed. The use of laparoscopic ultrasound could also localize concomitant abscesses and detect associated biliary tract pathology. PMID- 9348627 TI - Tension pneumothorax precluding laparoscopic repair of diaphragmatic hernia. AB - Pneumothorax can result from laparoscopic procedures in the abdomen. Usually, pneumothoraxes are mild and asymptomatic and do not require conversion to an open procedure. We report a case of tension pneumothorax that developed during the course of a laparoscopic repair of a diaphragmatic hernia. In this patient, the tension pneumothorax did not respond to conventional means of therapy and required conversion to a laparotomy. A large diaphragmatic hernia with communication between the peritoneal and pleural cavities may be a contraindication to minimally invasive laparoscopic procedures. PMID- 9348628 TI - Stones spilled during cholecystectomy: a long-term liability for the patient. AB - Within the last 2 years, an increasing number of case reports concerning stone spillage during laparoscopic cholecystectomy and its long-term consequences have been published. Recently three patients were treated for abscesses caused by spilled stones at our institution. One of them had the longest interval between cholecystectomy and abscess formation on record. Her abscess developed 20 years after open cholecystectomy. The second patient had been admitted with one of the few cases of cholelithopthysis reported after laparoscopic cholecystectomy. All three cases and their long history of recurrences clearly underline the necessity for open debridement and drainage with stone removal for definitive treatment of these patients. PMID- 9348629 TI - Is laparoscopic cholecystectomy recommended for large polypoid lesions of the gallbladder? PMID- 9348630 TI - Geriatric care in the late 1990s. AB - Geriatric health care remains a virtually untapped area in spite of changing public views and scientific knowledge on aging. The number of older dogs and cats will continue to grow into the next century. Owners of older dogs and cats are willing to invest in quality health care services. A geriatric health care program can expand veterinary services in a practice and is a natural extension of pediatric and adult maintenance programs. PMID- 9348631 TI - Endocrinopathies. Thyroid and adrenal disorders. AB - This article focuses on common adrenal and thyroid diseases in the geriatric patient consisting of hypothyroidism in the dog, hyperthyroidism in the cat, and hyperadrenocorticism in the dog to include clinical signs, diagnosis, and management. A brief section on hyperadrenocorticism in the cat, thyroid tumors in the dog, and pheochromocytoma in the dog and cat are also included. PMID- 9348632 TI - Chronic heart disease and its management. AB - Diseases of the cardiovascular system are a common cause of morbidity and mortality among geriatric canine and feline patients. Furthermore, diseases of the heart and vasculature often complicate the management of other abnormalities. This article contains a brief overview of the effects of aging on the circulatory system. It also contains a discussion of the cardiovascular disorders most frequently encountered in geriatric animals, namely, the feline myocardial disorders and feline hypertension and degenerative valvular disease, dilated cardiomyopathy, and cardiac in dogs. PMID- 9348633 TI - Chronic renal failure and its management and nephrolithiasis. AB - Chronic renal failure (CRF) is the most common form of renal disease in dogs and cats. Although CRF occurs in dogs and cats of all ages, it is commonly considered a disease of older animals, and the incidence increases with age. This article presents guide-lines for the diagnosis and conservative management of chronic renal failure in dogs and cats. Nephrolithiasis is uncommon in dogs and cats, accounting for less than 3% of all urinary calculi. The mineral composition of the renolith is important in formulation of therapeutic and preventive management strategies. This article briefly reviews the epidemiology, diagnosis, and management of nephrolithiasis in the dog and cat. PMID- 9348634 TI - Systemic hypertension and its management. AB - The pathophysiology of hypertension in dogs and cats, the methods available to monitor blood pressure, and the signs and treatment of hypertension are reviewed. Clinical signs of hypertension are usually referable to target organ damage, most notably in ophthalmic, renal, and cardiovascular tissues, which have a rich arteriolar supply. Blood pressure should be measured in any animal with renal disease, hyperthyroidism, hyperadrenocorticism, retinal detachment or hemorrhage, hyphema, or echocardiographically determined cardiac hypertrophy. All cats with acquired cardiac murmur should also be evaluated for hypertension. Antihypertensive medication should be administered if the indirect blood pressure in cats is consistently over 170/100 mmHg, or if the indirect blood pressure in dogs is greater than 180/100 mmHg. PMID- 9348635 TI - Gastrointestinal disease and its management. AB - Gastrointestinal (GI) diseases involving the alimentary tract and hepatobiliary system are common in geriatric dogs and cats. Inflammatory disorders predominate, but motility disturbances and degenerative lesions may also cause GI signs in affected animals. Treatment is directed at correction of the underlying cause and often requires tissue biopsy. The prognosis is good in many diseases with appropriate drug nutritional, and/or surgical therapy. PMID- 9348636 TI - Imaging abdominal masses. AB - Both radiography and ultrasound provide noninvasive imaging of suspected abdominal masses with minimal discomfort or risk for the geriatric patient. Radiography is more readily available and less expensive than ultrasonography, but contrast resolution is poor. Displacement of adjacent structures and addition of special contrast studies will provide clues to the possible organ of origin and extent of suspected abdominal masses. Cystic lesions can be differentiated from solid masses with ultrasound, but the appearance of focal abnormalities is not specific for any one disease process. Abdominal ultrasonography often provides the best diagnostic yield when used in combination with radiography and image-guided biopsy techniques. PMID- 9348637 TI - Musculoskeletal system. Joint and vertebral column diseases. AB - Owner complaints that refer to the musculoskeletal system are common in older dogs and cats. When the veterinarian is presented with these types of complaints, the differential lists include chronic intervertebral disk disease, diskospondylitis, degenerative joint disease, spondylosis with nerve root compression, joint/ ligament instability, and/or cancer. The diagnosis and management of some of these conditions is presented in detail with the general goal in mind that the older dog or cat is provided the best quality of life possible through good mobility along with being pain free. PMID- 9348638 TI - Common neurologic diseases of older animals. AB - Of the various neurologic diseases that affect dogs and cats, some are more often encountered in older animals. Physical diagnosis may be challenging, as multiple disease processes that may minic neurologic disease can be present in the same animal. A diligent, complete neurologic examination should lend for an accurate neuroanatomical diagnosis. Once a level of involvement is determined, knowledge of diseases affecting the particular area of the nervous system will provide for appropriate diagnosis, treatment, and prognosis. Although neurologic diseases are still often devastating, successful management of many of these diseases will afford better quality of life during the geriatric years. PMID- 9348639 TI - Pain and its management. AB - The management of pain in geriatric patients is a critical part of good veterinary care. The older patient poses unique issues in the selection of appropriate analgesic drugs due to altered drug absorption, metabolism and the frequent occurrence of underlying disease. Narcotics and NSAIDs are important analgesics that can be safely administered in geriatric patients provided that dosing amount and frequency are adjusted. Alternative analgesic techniques, such as nerve blocks, spinal analgesia and transcutaneous patches, offer a particularly useful source of analgesia in compromised geriatric patients. PMID- 9348640 TI - Ophthalmic disease and its management. AB - A variety of ocular disorders occur with increased frequency in aging patients. There are those, such as lens-induced uveitis, which simply represent the end stages of chronic disease. There are lesions of the orbit, eyelid, and uveal tract that epitomize the geriatric penchant for neoplasia. Calcific degeneration and endothelial dystrophy are among the disorders representing the degenerative side of the aging eye. Vision loss accompanies cataracts and retinal detachments. Though each of these disorders is potentially blinding, advances in medical and surgical management have improved the likelihood of lifelong vision in our veterinary patients. PMID- 9348641 TI - Ear disease and its management. AB - Tissue changes, diseases or factors predisposing to, bacteria and yeast associated with, diagnosis of, and management of otitis externa, media, and interna are discussed in this article. PMID- 9348642 TI - Geriatric behavioral problems. AB - Behavior problems in older pets may be due to many of the same causes as in younger pets. However, the effects of aging on the pet's body may cause a dramatic decline or deterioration in organ and sensory function, which may have a profound impact on the pet's behavior. A decline in cognitive function may also afflict older pets, in many instances due to the occurrence of Alzheimer's disease brain pathology, especially beta amyloid plaque formation. Correlation of geriatric behavior problems, therefore, entails first diagnosing and treating any underlying medical problems. Behavior modification techniques and drug therapy may then be required, with modifications and adjustments made to suit the specific needs of the older pet. PMID- 9348643 TI - Nutritional management. AB - This article describes nutritional assessment for geriatric patients and addresses some nutrient sensitive conditions common in older dogs and cats. The goal of completing a nutritional assessment is to identify the presence and significance of factors that put patients at risk for malnutrition. Dietary recommendations for geriatric patients should take into account the needs of the patient and client preferences as well as economics. Changes in feeding management should be considered a part of total patient management. As with any aspect of medical management, the patient should be reevaluated at appropriate intervals to assure achievement of desired results. PMID- 9348644 TI - Phototransduction and circadian clock pathways regulating gene transcription in higher plants. PMID- 9348645 TI - Apoptosis. PMID- 9348646 TI - Molecular basis for X-linked immunodeficiencies. PMID- 9348648 TI - The contribution of the mouse to advances in human genetics. PMID- 9348647 TI - Gene therapy of Duchenne muscular dystrophy. PMID- 9348649 TI - prune/Killer of prune: a conditional dominant lethal interaction in Drosophila. PMID- 9348650 TI - Chiasmata, crossovers, and meiotic chromosome segregation. AB - Meiotic recombination events are probably critical for the completion of several meiotic processes. In addition, recombination is likely to be involved in the events that lead up to synapsis of homologues in meiotic prophase. Recombination events that ultimately become resolved as exchanges are needed for the formation of chiasmata. Chiasmata maintain the association of paired homologues following loss of the synaptonemal complex and participate in the mechanism that signals that the bivalent has attached to the spindle in a bipolar orientation that will result in meiosis I disjunction. PMID- 9348651 TI - Molecular genetics of familial cardiomyopathies. PMID- 9348652 TI - The peripheral neuropathies and their molecular genetics. PMID- 9348653 TI - Tumor suppressor genes and human cancer. PMID- 9348654 TI - Genetic redundancy. PMID- 9348655 TI - Genetics of hybrid inviability in Drosophila. PMID- 9348656 TI - Regulation of bacterial gene expression by metals. PMID- 9348657 TI - Chromosome rearrangements in Neurospora and other filamentous fungi. AB - Knowledge of fungal chromosome rearrangements comes primarily from N. crassa, but important information has also been obtained from A. nidulans and S. macrospora. Rearrangements have been identified in other Sordaria species and in Cochliobolus, Coprinus, Magnaporthe, Podospora, and Ustilago. In Neurospora, heterozygosity for most chromosome rearrangements is signaled by the appearance of unpigmented deficiency ascospores, with frequencies and ascus types that are characteristic of the type of rearrangement. Summary information is provided on each of 355 rearrangements analyzed in N. crassa. These include 262 reciprocal translocations, 31 insertional translocations, 27 quasiterminal translocations, 6 pericentric inversions, 1 intrachromosomal transposition, and numerous complex or cryptic rearrangements. Breakpoints are distributed more or less randomly among the seven chromosomes. Sixty of the rearrangements have readily detected mutant phenotypes, of which half are allelic with known genes. Constitutive mutations at certain positively regulated loci involve rearrangements having one breakpoint in an upstream regulatory region. Of 11 rearrangements that have one breakpoint in or near the NOR, most appear genetically to be terminal but are in fact physically reciprocal. Partial diploid strains can be obtained as recombinant progeny from crosses heterozygous for insertional or quasiterminal rearrangements. Duplications produced in this way precisely define segments that cover more than two thirds of the genome. Duplication-producing rearrangements have many uses, including precise genetic mapping by duplication coverage and alignment of physical and genetic maps. Typically, fertility is greatly reduced in crosses parented by a duplication strain. The finding that genes within the duplicated segment have undergone RIP mutation in some of the surviving progeny suggests that RIP may be responsible for the infertility. Meiotically generated recessive-lethal segmental deficiencies can be rescued in heterokaryons. New rearrangements are found in 10% or more of strains in which transforming DNA has been stably integrated. Electrophoretic separation of rearranged chromosomal DNAs has found useful applications. Synaptic adjustment occurs in inversion heterozygotes, leading progressively to nonhomologous association of synaptonemal complex lateral elements, transforming loop pairing into linear pairing. Transvection has been demonstrated in Neurospora. Beginnings have been made in constructing effective balancers. Experience has increased our understanding of several phenomena that may complicate analysis. With some rearrangements, nondisjunction of centromeres from reciprocal translocation quadrivalents results in 3:1 segregation and produces asci with four deficiency ascospores that occupy diagnostic positions in linear asci. Three-to-one segregation is most frequent when breakpoints are near centromeres. With some rearrangements, inviable deficiency ascospores become pigmented. Diagnosis must then depend on ascospore viability. In crosses between highly inbred strains, analysis may be handicapped by random ascospore abortion. This is minimized by using noninbred strains as testers. PMID- 9348658 TI - Stark spectroscopy: applications in chemistry, biology, and materials science. AB - Stark spectroscopy has been applied to a wide range of molecular systems and materials. A generally useful method for obtaining electronic and vibrational Stark spectra that does not require sophisticated equipment is described. By working with frozen glasses it is possible to study nearly any molecular system, including ions and proteins. Quantitative analysis of the spectra provides information on the change in dipole moment and polarizability associated with a transition. The change in dipole moment reflects the degree of charge separation for a transition, a quantity of interest to a variety of fields. The polarizability change describes the sensitivity of a transition to an electrostatic field such as that found in a protein or an ordered synthetic material. Applications to donor-acceptor polyenes, transition metal complexes (metal-to-ligand and metal-to-metal mixed valence transitions), and nonphotosynthetic biological systems are reviewed. PMID- 9348659 TI - Infrared and Raman vibrational optical activity: theoretical and experimental aspects. AB - Advances in the field of vibrational optical activity (VOA) are reviewed over the past decade. Topics are surveyed with an emphasis on the theoretical and instrumental progress in both vibrational circular dichroism (VCD) and Raman optical activity (ROA). Applications of VOA to stereochemical and biological problems are reviewed, with a bias toward new kinds of experiments made possible by theoretical and instrumental advances. In the field of VCD, the most notable advances have taken place in the quality and size of ab initio calculations of VOA intensities and in the quality of step-scan Fourier transform instrumentation. For ROA, the most dramatic progress has occurred in the areas of theoretical formulation and high-throughput instrumentation. Applications of VOA now include all major classes of biological and pharmaceutical molecules. VOA's importance as a diagnostic tool will likely grow as the control of molecular chirality increases in research and industrial areas. PMID- 9348660 TI - Molecular structure and dynamics at liquid-liquid interfaces. AB - The structural, dynamical, and electrical properties of the interface between two immiscible liquids are described. The adsorption of solute molecules and the processes of ion transfer across the interface and of electron transfer at the interface are discussed. The microscopic perspective is emphasized by focusing on selected recent experimental results and on results obtained from molecular dynamics and Monte Carlo computer simulations. The validity of some of the existing models of the interface is examined. A proper account of the molecular structure of the interface is important for understanding solvation and charge transfer processes at the interface. PMID- 9348661 TI - Fast natural and magnetic circular dichroism spectroscopy. AB - The addition of circular or, more generally, elliptical polarization state detection to fast optical absorption spectroscopy can increase the amount of electronic and nuclear conformational information obtained about transient molecular species. To accomplish this, fast circular dichroism methods have emerged over the past decade that overcome the millisecond limit on time resolution associated with conventional modulation techniques and enable structural studies of excited states and kinetic intermediates. This article reviews techniques for time-resolved natural and magnetic circular dichroism spectroscopy covering the picosecond to millisecond time regimes and their applications, with particular emphasis on quasi-null ellipsometric techniques for nanosecond multichannel measurements of circular dichroism. Closely related quasi null polarimetric techniques for nanosecond optical rotatory dispersion and linear dichroism measurements are also discussed. PMID- 9348662 TI - "Strong" hydrogen bonds in chemistry and biology. AB - Hydrogen bonds are a key feature of chemical structure and reactivity. Recently there has been much interest in a special class of hydrogen bonds called "strong" or "low-barrier" and characterized by great strength, short distances, a low or vanishing barrier to hydrogen transfer, and distinctive features in the NMR spectrum. Although the energy of an ordinary hydrogen bond is ca 5 kcal mol-1, the strength of these hydrogen bonds may be > or = 10 kcal mol-1. The properties of these hydrogen bonds have been investigated by many experimental techniques, as well as by calculation and by correlations among those properties. Although it has been proposed that strong, short, low-barrier hydrogen bonds are important in enzymatic reactions, it is concluded that the evidence for them in small molecules and in biomolecules is inconclusive. PMID- 9348663 TI - Theory of protein folding: the energy landscape perspective. AB - The energy landscape theory of protein folding is a statistical description of a protein's potential surface. It assumes that folding occurs through organizing an ensemble of structures rather than through only a few uniquely defined structural intermediates. It suggests that the most realistic model of a protein is a minimally frustrated heteropolymer with a rugged funnel-like landscape biased toward the native structure. This statistical description has been developed using tools from the statistical mechanics of disordered systems, polymers, and phase transitions of finite systems. We review here its analytical background and contrast the phenomena in homopolymers, random heteropolymers, and protein-like heteropolymers that are kinetically and thermodynamically capable of folding. The connection between these statistical concepts and the results of minimalist models used in computer simulations is discussed. The review concludes with a brief discussion of how the theory helps in the interpretation of results from fast folding experiments and in the practical task of protein structure prediction. PMID- 9348664 TI - Classification of all putative permeases and other membrane plurispanners of the major facilitator superfamily encoded by the complete genome of Saccharomyces cerevisiae. AB - On the basis of the complete genome sequence of the budding yeast Saccharomyces cerevisiae, a computer-aided analysis was carried out of all members of the major facilitator superfamily (MFS), which typically consists of permeases with 12 transmembrane spans. Analysis of all 5885 predicted open reading frames identified 186 potential MFS proteins. Binary sequence comparison made it possible to cluster 149 of them into 23 families. Putative permease functions could be assigned to 12 families, the largest including sugar, amino acid, and multidrug transport. Phylogenetic clustering of proteins allowed us to predict a possible permease function for a total of 119 proteins. Multiple sequence alignments were made for all families, and evolutionary trees were constructed for families with at least four members. The latter resulted in the identification of 21 subclusters with presumably tightly related permease function. No functional clues were predicted for a total of 41 clustered or unclustered proteins. PMID- 9348665 TI - Organization and regulation of cbb CO2 assimilation genes in autotrophic bacteria. AB - The Calvin-Benson-Bassham cycle constitutes the principal route of CO2 assimilation in aerobic chemoautotrophic and in anaerobic phototrophic purple bacteria. Most of the enzymes of the cycle are found to be encoded by cbb genes. Despite some conservation of the internal gene arrangement cbb gene clusters of the various organisms differ in size and operon organization. The cbb operons of facultative autotrophs are more strictly regulated than those of obligate autotrophs. The major control is exerted by the cbbR gene, which codes for a transcriptional activator of the LysR family. This gene is typically located immediately upstream of and in divergent orientation to the regulated cbb operon, forming a control region for both transcriptional units. Recent studies suggest that additional protein factors are involved in the regulation. Although the metabolic signal(s) received by the regulatory components of the operons is (are) still unknown, the redox state of the cell is believed to play a key role. It is proposed that the control of the cbb operon expression is integrated into a regulatory network. PMID- 9348666 TI - Transposition and site-specific recombination: adapting DNA cut-and-paste mechanisms to a variety of genetic rearrangements. AB - In bacteria, two categories of specialised recombination promote a variety of DNA rearrangements. Transposition is the process by which genetic elements move between different locations of the genome, whereas site-specific recombination is a reaction in which DNA strands are broken and exchanged at precise positions of two target DNA loci to achieve determined biological function. Both types of recombination are represented by diverse genetic systems which generally encode their own recombination enzymes. These enzymes, generically called transposases and site-specific recombinases, can be grouped into several families on the basis of amino acid sequence similarities, which, in some cases, are limited to a signature of a few residues involved in catalysis. The well characterised site specific recombinases are found to belong to two distinct groups whereas the transposases form a large super-family of enzymes encompassing recombinases from both prokaryotes and eukaryotes. In spite of important differences in the catalytic mechanisms used by these three classes of enzymes to cut and rejoin DNA molecules, similar strategies are used to coordinate the biochemical steps of the recombination reaction and to control its outcome. This review summarises our current understanding of transposition and site-specific recombination, attempting to illustrate how relatively conserved DNA cut-and-paste mechanisms can be used to bring about a variety of complex DNA rearrangements. PMID- 9348667 TI - Karyotypic evolution of cells in culture: a new concept. AB - The Chapter summarizes peculiarities of karyotypic variability during establishment and long-term cultivation of permanent cell lines. A new concept on pathways of karyotypic evolution of cells in culture is put forward. A detailed description is presented of the author's original approach of cytogenetic analysis of cell lines provided for a principally new characteristic of the cell line: its generalized reconstructed karyotype (GRK). Its use as a criterion to evaluate authenticity, purity, and stability of cell lines is discussed. Based on analysis of the GRK, two stages of karyotype evolution of cell lines are revealed: establishment and stabilization, different in karyotypic variability of the cell population and in peculiarities of clone selection. Comparison of peculiarities of karyotypic variability of leukemic and tumor cells both in vitro and in vivo was made, and general regularities of their karyotypic evolution have been established, such as nonrandom changes in the number and structure of chromosomes and deletion of one of the sex chromosomes, as well as regularities characteristic only of cells in culture in most human and animal cell lines (at least 85%) of disomy on all autosomes. The rest of the cell lines, 15%, are characterized by either partial or total monosomies on certain autosomes during long-term cultivation. Three main compensatory mechanisms of maintaining viability of cell lines that have lost genetic material are discussed: polyploidization of the initial cell clone, amplification of oncogenes (predominantly of mys family), and extracopying of whole autosomes or of their fragments. PMID- 9348668 TI - Sucrose transport in higher plants. AB - Presumably due to its physicochemical properties, sucrose represents the major transport form of photosynthetically assimilated carbohydrates in plants. Sucrose synthesized in green leaves is transported via the phloem, the long distance distribution network for assimilates in order to supply nonphotosynthetic organs with energy and carbon skeletons. At least in Solanaceae, sugar export seems to be a tightly regulated process involving a number of specific plasma membrane proteins. Significant progress in this field was made possible by the recent identification of plasma membrane sucrose transporter genes. These carriers represent important parts of the long-distance transport machinery and can serve as a starting point to obtain a complete picture of long-distance sucrose transport in plants. A combination of biochemical studies of transporter properties together with expression and localization studies allow specific functions to be assigned to the individual proteins. Furthermore, the use of transgenic plants specifically impaired in sucrose transporter expression has provided strong evidence that SUT1 transporter function is required for phloem loading. Physiological analyses of these plants demonstrate that sucrose transporters are essential components of the sucrose translocation pathway at least in potato and tobacco. PMID- 9348669 TI - Interaction of cytoskeletal proteins with membrane lipids. AB - Rapid and significant progress has been made in understanding lipid/protein interactions involving cytoskeletal components and the plasma membrane. Covalent and noncovalent lipid modifications of cytoskeletal proteins mediate their interaction with lipid bilayers. The application of biophysical techniques such as differential scanning colorimetry, neutron reflection, electron spin resonance, CD spectroscopy, nuclear magnetic resonance, and hydrophobic photolabeling, allow various folding stages of proteins during electrostatic adsorption and hydrophobic insertion into lipid bilayers to be analyzed. Reconstitution of proteins into planar lipid films and liposomes help to understand the architecture of biological interfaces. During signaling events at plasma membrane interfaces, lipids are important for the regulation of catalytic protein functions. Protein/lipid interactions occur selectively and with a high degree of specificity and thus have to be considered as physiologically relevant processes with gaining impact on cell functions. PMID- 9348670 TI - Cell biology of autoimmune diseases. AB - Autoimmune diseases such as insulin-dependent diabetes mellitus, rheumatoid arthritis, and multiple sclerosis are common in the western world and are often devastating diseases which pose serious health problems. The key feature of such diseases is the development and persistence of inflammatory processes in the apparent absence of pathogens, leading to chronic breakdown of selected tissues. To date, no comprehensive explanation can be given for the onset or persistence of autoimmunity. As a rule, the chronic activation of helper T lymphocytes reactive against self proteins appears to be crucial for fueling the destructive autoimmune process, but why this occurs remains to be established. In this review, we present an overview on the rules that govern activation of T lymphocytes and on the factors that control it. The contribution of both genetic and environmental factors are discussed, clarifying that most autoimmune disease are of multifactorial origin. Special emphasis is given to the contribution of infectious events and the role of stress proteins in the process. In attempts to dissect the mechanisms involved in autoimmunity and to develop ways of blocking disease, experimental animal models are widely employed. We describe the various experimental models that exist for the study of multiple sclerosis, diabetes, and other autoimmune diseases and on the experience that has been gained in such models with experimental therapies to block the activation of self-reactive T lymphocytes. The lessons that can be drawn from these studies provide hope that continued efforts will lead to the successful development of antigen-specific strategies which block the development of autoimmunity also in humans. PMID- 9348671 TI - Multiple forms of tubulin: different gene products and covalent modifications. AB - Tubulin, the subunit protein of microtubules, is an alpha/beta heterodimer. In many organisms, both alpha and beta exist in numerous isotypic forms encoded by different genes. In addition, both alpha and beta undergo a variety of posttranslational covalent modifications, including acetylation, phosphorylation, detyrosylation, polyglutamylation, and polyglycylation. In this review the distribution and possible functional significance of the various forms of tubulin are discussed. In analyzing the differences among tubulin isotypes encoded by different genes, some appear to have no functional significance, some increase the overall adaptability of the organism to environmental challenges, and some appear to perform specific functions including formation of particular organelles and interactions with specific proteins. Purified isotypes also display different properties in vitro. Although the significance of all the covalent modification of tubulin is not fully understood, some of them may influence the stability of modified microtubules in vivo as well as interactions with certain proteins and may help to determine the functional role of microtubules in the cell. The review also discusses isotypes of gamma-tubulin and puts various forms of tubulin in an evolutionary context. PMID- 9348672 TI - Translocation of proteins across the endoplasmic reticulum membrane. AB - Secretory protein biogenesis begins with the insertion of a preprotein into the lumen of the endoplasmic reticulum (ER). This insertion event, known as ER protein translocation, can occur either posttranslationally, in which the preprotein is completely synthesized on cytosolic ribosomes before being translocated, or cotranslationally, in which membrane-associated ribosomes direct the nascent polypeptide chain into the ER concomitant with polypeptide elongation. In either case, preproteins are targeted to the ER membrane through specific interactions with cytosolic and/or ER membrane factors. The preprotein is then transferred to a multiprotein translocation machine in the ER membrane that includes a pore through which the preprotein passes into the ER lumen. The energy required to drive protein translocation may derive either from the coupling of translation to translocation (during cotranslational translocation) or from ER lumenal molecular chaperones that may harness the preprotein or regulate the translocation machinery (during posttranslational translocation). PMID- 9348673 TI - The influence of different timbre attributes on the perceptual segregation of complex-tone sequences. AB - Spectral factors such as differences in harmonic content are powerful cues in the perceptual organization of tone sequences. Temporal features such as rise time, however, have been shown to be poor cues [W. M. Hartmann and D. Johnson, Mus. Perc. 9, 155-184 (1991)]. The relative influence of these timbral features on perceptual segregation was investigated. Complex tones were sequenced in a repeating ABA- "gallop" format, under four conditions in which tones A and B had the same or different timbres as defined by differences in numbers of harmonics and temporal-envelope features. A sequence started with A and B tones at the same F0. The F0 difference between A and B then increased over the course of a trial, until a listener terminated the trial indicating perceptual segregation into sub sequences comprising A and B tones, respectively. The F0 difference required to reach this crossover point of segregation provided a measure of the efficacy of stimulus features of A and B as cues for perceptual organization. Sequences combining differences in harmonic structure and temporal envelope required the smallest F0 change for segregation. Sequences of tones with the same harmonic structure and temporal envelope required larger changes in F0, while the other conditions fell in the middle of this range. The F0-tracking method used in this study facilitates measurement of the relative contribution of different stimulus features to stream segregation. It also holds potential as a tool using the point of segregation as a measure of the magnitude of timbre differences brought about by different physical features of sounds. PMID- 9348674 TI - Finite difference predictions of P-SV wave propagation inside submerged solids. I. Liquid-solid interface conditions. AB - A finite difference scheme has been developed to analyze internal strains in submerged elastic solids of irregular geometry subjected to ultrasonic wave sources that simulate a clinical lithotripter. In part I of this paper, the finite difference formulation that accounts for arbitrary liquid-solid interfaces is presented and sample numerical results are discussed. Two different methods for discretizing the liquid-solid interface conditions are developed. The first treats the interface conditions explicitly. The second integrates the heterogeneous wave equations across the interface using the divergence theorem. Both schemes account for varying grid sizes and give similar results for a test problem consisting of a radially diverging source incident on the rectangular solid. The time sequence obtained numerically for strain at the center of a rectangular solid matches well with the experimental results [S. M. Gracewski et al., J. Acoust. Soc. Am. 94, 652-661 (1993)] in terms of the arrival times and the relative amplitudes of the peaks. In addition, strain contours are plotted to visualize the propagation of P (longitudinal) and S (shear vertical) waves inside a circular solid. The reflection from the concave back surface of the circular solid has a focusing effect with the subsequent formation of focal zones, known as caustics, where peak strains occur. In part II of this paper, the finite difference scheme is used to study the effects of geometry changes on the internal stresses and caustics predicted in model stones subjected to lithotripter pulses. PMID- 9348675 TI - Finite difference predictions of P-SV wave propagation inside submerged solids. II. Effect of geometry. AB - To understand better direct stress wave contributions to stone fragmentation during extracorporeal shock wave lithotripsy (ESWL), the numerical formulation developed in part I is applied to study the time evolution of stress wave fields produced inside submerged isotropic elastic solids having irregular geometries. Cut spheres are used to model stones that have already had an initial fracture. Ellipses are used to approximate other deviations from a spherical geometry. The propagation and focusing of the longitudinal (P) and shear (S) wave fronts are visualized by presenting internal strain contours. Internal strain measurements are obtained from strain gauges embedded inside plaster specimens to confirm the focusing effect obtained from the concave back surfaces of the stones. Fragmentation experiments indicate damage caused by spalling and direct stress wave focusing as well as a front surface pit presumably created by cavitation activity. PMID- 9348676 TI - Implementation of an active headset by using the H infinity robust control theory. AB - This paper presents a methodology for implementing an active headset by using H infinity robust control theory. The adopted structure is feedback tracking control. Performance, stability, and robustness of the closed-loop system have been taken into account in the design procedure by using a general framework of the H infinity theory. The resultant controller is realized on the basis of operational amplifier circuitry. Experiments are conducted to test the developed headset. The result shows that the headset achieves broadband attenuation up to approximately 15 dB in the band 200-800 Hz. The design considerations indicated in the experimental result are also addressed. PMID- 9348677 TI - Individual differences in susceptibility to the "irrelevant speech effect". AB - Individual differences in objective effects of noise on performance were analyzed with respect to their distribution, temporal stability, and the precision of measurement to be attained. Seventy-two subjects had to memorize sequences of visually presented digits while being exposed to one of three auditory background conditions which were randomly mixed on a trial-by-trial basis: (1) foreign speech; (2) pink noise; and (3) silence. Individual "irrelevant speech effects," operationalized by the difference in recall errors under speech and in silence, were normally distributed over a wide range extending from slight facilitation to severe disruption. When 25 subjects repeated the experiment after four weeks, the individual differences were replicated with a reliability of rtt = 0.45. Internal consistency, a measure of the precision with which individual effects can be measured in a single session, was moderate (alpha = 0.55). However, both retest, and consistency coefficients are severely attenuated by the use of (sound-minus silence) difference scores, the reliability of which is bound to be considerably lower than that of the original error scores whenever these are correlated. Given that the original error rates in a specific auditory condition can be determined with reliabilities approaching 0.85, it may be concluded that individual performance decrements due to noise can be reliably measured in the "irrelevant speech" paradigm. Self-report measures of noise susceptibility collected to explore potential sources of the large inter-individual variation exhibited only weak relationships with the objectively measured noise effects: Subjects were quite inaccurate in assessing their individual impairment in the three auditory conditions, and a questionnaire-based measure of general noise sensitivity only accounted for a small portion of the variance in objectively measured performance decrements, although in both cases the predictive relationship was much stronger in female than in male subjects. PMID- 9348678 TI - A time domain binaural model based on spatial feature extraction for the head related transfer function. AB - A complex-valued head-related transfer function (HRTF) can be represented as a real-valued head-related impulse response (HRIR). The interaural time and level cues of HRIRs are extracted to derive the binaural model and also to normalize each measured HRIR. Using the Karhunen-Loeve expansion, normalized HRIRs are modeled as a weighted combination of a set of basis functions in a low dimensional subspace. The basis functions and the space samples of the weights are obtained from the measured HRIR. A simple linear interpolation algorithm is employed to obtain the modeled binaural HRIRs. The modeled HRIRs are nearly identical to the measured HRIRs from an anesthetized live cat. Typical mean square errors and cross-correlation coefficients between the 1816 measured and modeled HRIRs are 1% and 0.99, respectively. The real-valued operations and linear interpolating in the model are very effective for speeding up the model computation in real-time implementation. This approach has made it possible to simulate real free-field signals at the two eardrums of a cat via earphones and to study the neuronal responses to such a virtual acoustic space (VAR). PMID- 9348679 TI - Influence of contralateral noise on distortion product latency in humans: is the medial olivocochlear efferent system involved? AB - To test the hypothesis of temporal modifications of cochlear responses when medial efferents are activated, otoacoustic emission latencies were estimated in 16 normal human subjects, in the presence and absence of a contralateral broadband noise, using measurements of the phase of the 2f1-f2 distortion product (group latency method). Significant decrease in the latency of lower frequency (0.8-2.7 kHz) emissions was found in the presence of increasing levels of contralateral sound, and this effect disappeared when the primary-tone levels increased to 60 dB SPL. To ensure that effects were not attributable to mechanisms involving middle ear structures, susceptible to activation by contralateral sound, latency measures were performed in seven subjects whose efferents were severed during a vestibular neurotomy and in two subjects with paralyzed stapedial muscle. Results in patients were compared to those obtained in three surgical control patients with intact efferent bundle, and in eight other normal subjects. All the subject groups exhibited a decrease in latency under contralateral sound except the patients with the severed efferent system who showed increased latencies. PMID- 9348680 TI - Effects of electrode configuration on psychophysical strength-duration functions for single biphasic electrical stimuli in cats. AB - The interface between electrode and neural target tissue is thought to influence certain characteristics of neural and behavioral responses to electrical stimulation of the auditory system. At present, the biophysical properties of this interface are not well understood. Here the effects of biphasic phase duration and electrode configuration on psychophysical threshold in response to electrical stimulation in cats are described. Five cats were trained to respond to acoustic stimuli using food as a reward in an operant reinforcement paradigm. After training, the animals were unilaterally deafened and implanted with a multicontact intracochlear electrode array. Thresholds for single presentations of biphasic current pulses were measured as a function of phase duration and electrode arrangement. Statistical analyses of the data indicated that strength duration function slopes between 200 and 1600 microseconds/phase were significantly different for the different electrode configurations and, overall, were unrelated to the absolute level of the strength-duration function (i.e., were independent of absolute threshold). For all subjects, the slope of this function for intermediate pulse durations was dependent on electrode configuration and most shallow for radial-bipolar configurations (-3.4 dB/doubling), was steepest for monopolar arrangements (-5.9 dB/doubling), and was intermediate for longitudinal-bipolar pairings. (-4.4 dB/doubling). Slopes for both shorter and longer phase duration stimuli were not significantly different. The underlying mechanisms for these effects may include, or be a combination of altered electrical field patterns, integrated activity across multiple fibers, and stochastic behavior of individual auditory neurons to electrical stimulation. PMID- 9348681 TI - Model calculations of the effects of wide-band inhibitors in the dorsal cochlear nucleus. AB - In two previous papers [Reed and Blum, J. Acoust. Soc. Am. 97, 425-438 (1995), Blum et al., J. Acoust. Soc. Am. 98, 181-191 (1995)] a computational model for signal processing in the dorsal cochlear nucleus (DCN) was developed. In those modelling studies, stellate cells inhibited only type II cells. In this study, the effect of including wide-band inhibitory (WBI) connections from stellate cells to type IV cells, as proposed by Nelken and Young [J. Neurophysiol. 71, 2446-2462 (1994)], is examined. Inclusion of the WBI connections improves the fit to the experimental pure tone response maps for both the "standard" and "non standard" cells examined by Spirou and Young [J. Neurophysiol. 66, 1750-1768 (1991)]. Thus, these modelling studies support the hypothesis of Nelken and Young [J. Neurophysiol. 71, 2446-2462 (1994)]. The degree of improvement is greatest for cells with prominent upper inhibitory sidebands. The qualitative features of the pure tone response map and the theoretical model allow one to deduce the probable frequency bias of the type II to type IV and stellate to type IV connections. PMID- 9348682 TI - Two-tone rate suppression boundaries of cochlear ganglion neurons in chickens following acoustic trauma. AB - The purpose of the present study was to examine the effects of acoustic trauma and hair cell loss and regeneration on the two-tone rate suppression (TTRS) boundaries of cochlear ganglion neurons in chickens. Chickens were exposed for 48 hours to a 525-Hz, 120-dB SPL tone which destroyed the hair cells and tectorial membrane in a crescent-shaped patch along the abneural side of the basilar papilla. Afterwards, TTRS boundaries were recorded from cochlear ganglion neurons at 0-1, 5, 14, and 28 days postexposure. Acoustic trauma reduced the percentage of neurons with TTRS boundaries below CF (TTRSb) (52.6% to 8.2%) and above CF (TTRSa) (88.4% to 46.6%). In addition, the exposure reduced TTRS boundary slopes, elevated best suppression threshold (BST), and increased the frequency separation between the tips of the TTRS boundaries and CF. All the TTRS measures started to recover by 5 days postexposure and by 14 days and 28 days postexposure, most measures had recovered to normal levels. However, the BST, TTRS slopes, and the frequency separation of TTRSb boundaries from CF were still slightly abnormal near the exposure frequency. In addition, the percentage of neurons with TTRS below CF was reduced significantly. The partial recovery of TTRS boundaries is presumably due to the regeneration of hair cells and the lower honeycomb layer of the tectorial membrane. The residual TTRS deficits observed 28 days postexposure were most closely associated with the missing upper fibrous layer of the tectorial membrane. PMID- 9348683 TI - Multidimensional scaling of complex sounds by school-aged children and adults. AB - A paired-comparisons procedure was used to evaluate the processing of complex, nonspeech sounds by 7- and 10-year-old children and adults. Stimuli were brief duration and included pure tones, harmonic complexes, and bands of noise. From their similarity ratings, a three-dimensional multidimensional scaling solution was derived. Results suggested that listeners classified the stimuli into clusters based upon periodicity and the number of spectral peaks. Within each cluster, the stimuli were ordered according to frequency. Because individual differences in the overall weightings of features were large, separate solutions were derived for two subgroups of listeners, formed based upon similarities in the pattern of dimension weights obtained in the group analysis. One subgroup, for whom the full group analysis captured a large proportion of the variance in the ratings, included the adults, many of the 10-year-olds, and a few of the 7 year-olds. The solution derived for this subgroup suggested that spectral and temporal information were weighted equally and integrated into all dimension weights. Frequency information was coded but given less weight and was not used for stimulus classification. A second subgroup of listeners included most of the 7-year-old and some of the 10-year-old children. Examination of their data suggested that they relied heavily on an analysis of the signals according to periodicity as was reflected in their temporal fine structure. Also encoded but of lesser importance were aspects of spectral shape and absolute frequency. PMID- 9348684 TI - Discrimination of changes in the spectral shape of noise bands. AB - Discrimination experiments were performed for a change in the spectral shape of noise bands. The subject's task was to discriminate noise bands with a positive spectral slope from those with a negative spectral slope. Thresholds were measured at several bandwidths and center frequencies, as well as for several noise samples. Experiments were performed while roving the overall intensity. At a fixed center frequency of 1 kHz, sensitivity was best for bandwidths of 3-6 semitones (ST). At larger bandwidths, thresholds increased only slowly. At a fixed bandwidth of 1 ST, threshold hardly changed as a function of the center frequency. At a fixed bandwidth of 58 Hz, threshold was lowest near 500-1000 Hz. Model calculations show that the EWAIF model [Feth, Percept. Psychophys. 15, 375 378 (1974)] can account for the present results if the signal's bandwidth does not exceed 1 ST. The IWAIF model [Anantharaman et al., J. Acoust. Soc. Am. 94, 723-729 (1993)] can account for the present results only if the signal's bandwidth is smaller than 1 ST but larger than about 25 Hz. Results obtained with broadband signals could be described only qualitatively with the multichannel model [Durlach et al., J. Acoust. Soc. Am. 80, 63-72 (1986)]. Then, the model needs the assumption that either the output of the different frequency bands cannot be optimally combined, or that only two bands are used in the discrimination process. The present results are compared with those obtained with two-tone complexes measured under identical conditions [Versfeld and Houtsma, J. Acoust. Soc. Am. 98, 807-816 (1995)]. PMID- 9348685 TI - Coherence masking protection in brief noise complexes: effects of temporal patterns. AB - Three experiments examined listeners' thresholds for classifying the pitch of a target signal in a masking noise when it was presented alone as compared to when it was presented with a "cosignal." The target signal was a narrow band of noise centered on either 375 or 625 Hz and the masker was noise low-pass filtered at 1000 Hz. The cosignal provided no information about the pitch of the target signal but could potentially combine with it to form an auditory object; it was spectrally well separated from the target signal, consisting of a band of noise ranging from 2200 to 2900 Hz. Experiment 1 showed that identification thresholds were lower when the target signal was paired with the cosignal than when it was presented alone if the onsets and offsets of the target signal and cosignal were temporally synchronous. This is an instance of "coherence masking protection," a phenomenon that has previously been established in the perception of vowels [P.C. Gordon, Percept. Psychophys, 59, 232-242 (1997)]. The effect disappears when the cosignal leads and lags the target signal by short durations, a finding that also matches that observed previously with vowels. The finding that temporal relations between the components of a stimulus have similar effects on the perception of nonspeech noise complexes and speech sounds suggests that speech perception makes use of general auditory mechanisms for perceptual integration of this sort. Experiments 2 and 3 examine further the role of temporal relations between the onsets and offsets of the target signal and the cosignal in producing coherence masking protection. The results show that either onset synchrony or offset synchrony is sufficient to produce the effect when the cosignal is of greater duration than the target signal, but that only onset synchrony produces the effect when the target signal has greater duration than the cosignal. This pattern indicates that the target signal and cosignal do not contribute equally to the formation of auditory objects. PMID- 9348686 TI - The role of spread excitation and suppression in simultaneous masking. AB - This experiment was intended to clarify the relative role of spread of excitation and suppression in simultaneous masking, for masker frequencies just below and well below the signal frequency. The experiment had two stages. In stage 1, growth-of-masking functions were measured in simultaneous masking for a 2200-Hz sinusoidal signal and a sinusoidal masker with frequency of either 1800 Hz or 500 Hz. Straight lines fitted to these data were used to determine masker levels that would give 10, 20, and 30 dB of masking. In stage 2, thresholds for detecting a brief 2200-Hz signal were measured using forward masking. It was reasoned that the threshold of the signal would give an indication of the amount of excitation evoked by the masker in the frequency region of the signal. Three forward maskers were used: (1) a 2200-Hz sinusoid at 10, 20, or 30 dB sensation level (SL); (2) a 2200-Hz sinusoid at the same levels as in (1) together with a sinusoid with frequency 500 or 1800 Hz at a level just sufficient to mask the 2200-Hz sinusoid. We refer to this as the "combined masker," (3) a 500-Hz or 1800-Hz sinusoid at the same levels as in (2) above. The 1800-Hz combined masker produced slightly less forward masking than the 2200-Hz masker (1), which might be explained in terms of suppression or as perceptual cueing. Both the 1800-Hz combined masker and the 1800-Hz component alone (3) gave significant amounts of forward masking (up to 18 dB), indicating that these maskers produced substantial excitation at 2200 Hz. This is consistent with the idea that the simultaneous masking of the 2200-Hz component in stage 1 was produced by spread of excitation rather than by suppression. The 500-Hz combined masker produced much less forward masking than the 2200-Hz component alone, indicating strong suppression of the 2200-Hz component of the combined masker by the 500-Hz component. However, both the 500 Hz combined masker and the 500-Hz component alone produced some forward masking. This is not consistent with the idea that masking of the 2200-Hz component in stage 1 (simultaneous masking) was produced solely by suppression. PMID- 9348687 TI - A probe-signal study of auditory discrimination of complex tones. AB - The present experiments used an analogue of the probe-signal method of Greenberg and Larkin [J. Acoust. Soc. Am. 44, 1513-1523 (1968)] to investigate the extent to which listeners direct attention to a particular spectral region when discriminating complex tones. The listeners' task was to discriminate between two seven-component complex tones on the basis of an increment in the level of a single component. On two-thirds of trials the increment was achieved by adding a fixed primary signal to one component of the complex. The primary-signal trials were relatively easy and were intended to cue listeners to attend to the component to which the primary was added. On the remaining trials a smaller probe signal was added either to the cued component, or to one of three other components. The results of the first experiment, in which the complex tones had a flat spectrum, showed significantly better performance for probe signals applied to the cued component compared to the other three components. To control for the possibility that the observed pattern of results was due to the use of timbral cues, a second experiment was conducted in which the spectral profile of the tones was randomized between trials. The results for the second experiment were similar to those of the first experiment and are consistent with the idea that listeners were focusing attention on the spectral region defined by the primary signal. PMID- 9348688 TI - Identification of multidimensional stimuli containing speech cues and the effects of training. AB - The purpose of this study was to determine how listeners with normal hearing make use of cues from multiple, independent stimulus dimensions when classifying synthesized stimuli containing acoustic cues found in speech. Listeners classified synthesized stimuli that differed on three independent and discriminable dimensions. The three dimensions of the stimuli included: (1) the fricative spectrum center frequency; (2) the slope of the frequency transition; and (3) the duration of the temporal gap. Each of the three dimensions could take on one of three possible values. Twenty-seven stimuli were synthesized using all possible combinations of the values on the three dimensions. Multidimensional scaling of paired-comparison similarity judgments confirmed the existence of three perceptual dimensions. Listeners were then trained to classify three exemplar stimuli as "circle," "triangle," and "square," respectively. Following this training, subjects classified all 27 stimuli as "circle," "triangle," and "square." From this it was determined how each listener's attention was distributed among the dimensions. Results indicated the listeners did not pay equal attention to the three stimulus dimensions. In addition, it was demonstrated that it was possible to train a listener to attend to a dimension that was not previously used to classify the stimuli. PMID- 9348689 TI - Auditory profile analysis: is there perceptual constancy for spectral shape for stimuli roved in frequency? AB - The ability to discriminate between two spectral shapes was investigated using a one-interval, two-alternative forced choice procedure. Each stimulus consisted of seven components equally spaced on a logarithmic frequency scale (frequency ratio of 1.2 between adjacent components). The two spectral shapes were produced by first generating a complex each of whose components was equally loud (when heard in isolation) and then increasing the level of either the third or the fifth component by a certain amount. To render pitch an unusable cue, the center frequencies of the stimuli were randomly varied between presentations (maintaining the ratio between adjacent components) over a range of about three octaves (279-2074 Hz). The data indicate that listeners were not able to distinguish between the two types of profiles even for very large increments in level of the third and the fifth components. The results suggest that there is no perceptual constancy for this type of spectral shape for stimuli roved over a sufficiently large frequency range. The spectral shape cannot be perceived independently of stimulus frequency. The results are related to the multi-channel model applied in profile analysis tasks. PMID- 9348690 TI - Perceptual segregation of a harmonic from a vowel by interaural time difference and frequency proximity. AB - The five experiments reported here examine the conditions under which sounds differing in their interaural time difference (ITD) are segregated for the purposes of perceiving a vowel's identity. Experiment 1 confirms previous findings that (i) a difference in ITD provides only a very weak cue for segregating a vowel's 500-Hz harmonic from the remainder of an isolated vowel; (ii) embedding the harmonic in a series of 500-Hz tones produces some segregation, which is enhanced if the harmonic and the tones differ in ITD from the rest of the vowel; and (iii) when these latter sounds are presented in the same block as isolated vowels, they facilitate segregation of the harmonic by ITD in the isolated vowels. The subsequent experiments show that this last effect, across-trial facilitation, is only produced by sounds which cue both the frequency and the ITD of the harmonic; either alone is insufficient. We also show that: (i) a single cue tone at the frequency of the harmonic is sufficient to facilitate the use of ITD in grouping; (ii) sequential organization by frequency proximity dominates over sequential organization by ITD when simultaneous sound sources are present; and (iii) the effectiveness of a cue tone can be abolished by capturing it into a synchronous harmonic complex. The experiments clarify the conditions under which ITDs contribute to the segregation of simultaneous sounds. PMID- 9348691 TI - The effect of head rotations on vertical plane sound localization. AB - Current understanding gives predominant weight to stationary cues for auditory localization. Two experiments were conducted to investigate the possible existence of a dynamic cue. The first experiment involved localization of concealed sources in the upper median vertical plane (MVP) and showed, as expected, that elevation was not detectable with motionless listening when high frequency energy was absent or when normal pinna function was distorted. Elevation under such conditions did become detectable with horizontal head rotations, provided low-frequency energy was present in the signal. This indicates that the basis of the dynamic cue is variation in the rate of transformation of low-frequency interaural time/phase differences. The second experiment involved localization of sources arrayed throughout upper and lower regions of the MVP and in the left lateral vertical plane (LVP); it showed that upper hemisphere sources can be distinguished somewhat from those in the lower hemisphere, even in motionless listening conditions, but more so with rotation. The greatest benefit for localization from rotation of the head appears to be gained for sources positioned in the front MVP. PMID- 9348692 TI - Dichotic beats of mistuned consonances. AB - The beats of mistuned consonances (BMCs) result from the presentation of two sinusoids at frequencies slightly mistuned from a ratio of small integers. Several studies have suggested that the source of dichotic BMCs is an interaction within a binaural critical band. In one case the mechanism has been explained as an aural harmonic of the low-frequency tone (f1) creating binaural beats with the high-frequency tone (f2). The other explanation involves a binaural cross correlation between the excitation pattern of f1 and the contralateral f2- occurring within the binaural critical band centered at f2. This study examined the detection of dichotic BMCs for the octave and fifth. In one experiment with the octave, narrow-band noise centered at f2 was presented to one ear along with f1. The other ear was presented with f2. The noise was used to prevent interactions in the binaural critical band centered at f2. Dichotic BMCs were still detected under these conditions, suggesting that binaural interaction within a critical band does not explain the effect. Localization effects were also observed under this masking condition for phase reversals of tuned dichotic octave stimuli. These findings suggest a new theory of dichotic BMCs as a between channel phase effect. The modified weighted-image model of localization [Stern and Trahiotis, in Auditory Physiology and Perception, edited by Y. Cazals, L. Demany, and K. Horner (Pergamon, Oxford, 1992), pp. 547-554] was used to provide an explanation of the between-channel mechanism. PMID- 9348693 TI - Auditory localization, detection of spatial separateness, and speech hearing in noise by hearing impaired listeners. AB - In two groups, one with sensorineural and the other with conductive-mixed hearing loss, measures were made of single-source localization and speech intelligibility in both spatially separate and nonseparate noise. There was also a test for detecting when two sounds came from the same location or from separate ones. Localization test results confirmed earlier findings, namely, disruption of vertical plane ability generally, and a further, particular disturbance to horizontal plane localization in the conductive-mixed group. Compared with a normal control group, there were only slight signs of benefit from separation of speech and noise in the region lateral to the listener, and virtually none in the frontal region. The new test, spatial separateness, had elements in common with both of the other tests, and links were observed from localization to separateness detection, and from separateness to benefit from separation of speech and noise. Localization was also related to speech hearing in nonspatially separated noise. PMID- 9348694 TI - The development of speech adaptation to an artificial palate. AB - An investigation of adaptation to palatal modification in [s] production was conducted using acoustic and perceptual analyses. The experiment assessed whether adaptation would occur subsequent to a brief period of intensive, target-specific practice. Productions of [sa] were elicited at five time intervals, 15 min apart, with an artificial palate in place. Between measurement intervals, subjects read [s]-laden passages to promote adaptation. Results revealed improvement in both acoustic and perceptual measures at the final time interval relative to the initial measurement period. Interestingly, the data also suggested changes to normal (unperturbed) articulation patterns during the same interval. Results are discussed in relation to the development of speech adaptation to a structural modification of the oral cavity. PMID- 9348695 TI - Acoustic correlates of English and French nasalized vowels. AB - Acoustic analysis of nasalized vowels in the frequency domain indicates the presence of extra peaks: one between the first two formants with amplitude P1 and one at lower frequencies, often below the first formant, with amplitude P0. The first-formant amplitude A1 is also reduced relative to its amplitude for an oral vowel. These acoustic characteristics can be explained by speech production theory. The objective of this study was to determine the values for the acoustic correlates A1-P1 and A1-P0 (dB) for quantifying nasalization. They were tested as measures of nasalization by comparing vowels between nasal consonants and those between stop consonants for English speakers. Also, portions of nasal vowels following a stop consonant were compared for speakers of French, which makes a linguistic distinction between oral and nasal vowels. In the analysis of English, the mean difference of A1-P1 measured in oral vowels and nasalized vowels had a range of 10 dB-15 dB; the difference of A1-P0 had a range of 6 dB-8 dB. In the study of French, the difference of A1-P1 measured between the least-nasalized portion and the most-nasalized portion of the vowel had a range of 9 dB-12 dB; for A1-P0, the difference ranged between 3 dB and 9 dB. In order to obtain an absolute acoustic measure of nasalization that was independent of vowel type, normalized parameters were calculated by adjusting for the influence of the vowel formant frequencies. PMID- 9348696 TI - Correlation dimension of electroglottographic data from healthy and pathologic subjects. AB - This paper considers the effects of nonstationarity, noise, and finite data sets on the estimation of the correlation dimension of time-series data characterizing the vibratory behavior of the vocal folds. The electroglottographic signal of sustained /a/ phonations from 10 healthy subjects and 20 subjects with vocal fold pathologies were used to reconstruct the state space in successively higher embeddings using the method of lags. The dimension values were calculated from the scaling region (the level area of the slope plots) which did not increase for higher embeddings. Reasonably defined scaling regions were found in all of the data from the healthy subjects and from five of the pathologic subjects, with values saturating between the first and second embeddings. The EGG data from those five pathologic subjects were nearly periodic. From the remaining 15 subjects, the scaling regions were highly constricted with nonconstant slopes, so that dimension values could not be confidently estimated. The results suggest that the correlation dimension is a highly subjective measure which is not usefully applied to abnormal EGG data. It is recommended that, if used, correlation dimension statistics need to be presented cautiously, and graphical presentation of the data should be included. PMID- 9348697 TI - Timing of pitch movements and accentuation of syllables in Dutch. AB - In this study, the relation between the timing of a rising or falling pitch movement and the syllable it accentuates is investigated. The five-syllable utterance /mamamamama/ was provided with a relatively fast rising or falling pitch movement. The timing of the movement was systematically varied and Dutch subjects were asked to indicate which syllable they perceived as accented. In order to find out where in the pitch movement the cue which induces the percept of accentuation is located, the duration of the pitch movement was varied. In order to find out which segments of the utterance this characteristic is linked to, the duration of the /m/ was varied. The results showed that the percept of accentuation is induced by a change in pitch at the start of the movement. The moment at which the course of pitch starts to change significantly determines which syllable is perceived as accented. If this moment lies some tens of milliseconds before the P-center, i.e., the perceptual moment of occurrence of the syllable, the preceding syllable is perceived as accented. For a rise, a high accent is perceived; for a fall, a low accent. If the pitch change occurs after this moment, the syllable with this P-center is perceived as accented. For the rise, a low accent is then perceived; for the fall, a high accent. This will be discussed in the light of earlier research on accentuation and of theoretical knowledge about pitch accents. PMID- 9348698 TI - Speech intelligibility as a function of the number of channels of stimulation for signal processors using sine-wave and noise-band outputs. AB - Vowels, consonants, and sentences were processed through software emulations of cochlear-implant signal processors with 2-9 output channels. The signals were then presented, as either the sum of sine waves at the center of the channels or as the sum of noise bands the width of the channels, to normal-hearing listeners for identification. The results indicate, as previous investigations have suggested, that high levels of speech understanding can be obtained using signal processors with a small number of channels. The number of channels needed for high levels of performance varied with the nature of the test material. For the most difficult material--vowels produced by men, women, and girls--no statistically significant differences in performance were observed when the number of channels was increased beyond 8. For the least difficult material- sentences--no statistically significant differences in performance were observed when the number of channels was increased beyond 5. The nature of the output signal, noise bands or sine waves, made only a small difference in performance. The mechanism mediating the high levels of speech recognition achieved with only few channels of stimulation may be the same one that mediates the recognition of signals produced by speakers with a high fundamental frequency, i.e., the levels of adjacent channels are used to determine the frequency of the input signal. The results of an experiment in which frequency information was altered but temporal information was not altered indicates that vowel recognition is based on information in the frequency domain even when the number of channels of stimulation is small. PMID- 9348699 TI - Development and evaluation of a German sentence test for objective and subjective speech intelligibility assessment. AB - A German sentence test was developed which is comprised of 20 test lists of ten sentences each. The test corpus is a selection from sentences for speech quality evaluation recorded with a male unschooled speaker. Performance-intensity curves were measured for each individual sentence in a speech-simulating babble noise with a total of 40 normal-hearing listeners. Based on these data and the phonemic transcription of the 200 sentences selected from the underlying speech corpus, 20 test lists were composed using a numerical optimization process. These 20 test lists are highly equivalent with respect to their performance-intensity curves, the number of words within each test list, the number of phonemes within each test list, and approximately the frequency distribution of the phonemes which approximates the phoneme frequency distribution of the German language. The equivalence of the respective performance-intensity curves was demonstrated in an independent experiment with 20 normal-hearing listeners. In addition, a comparison was performed between the "objective" intelligibility measurements and two "subjective" speech intelligibility rating methods employing the same materials. As a result, both subjective assessment procedures correlate highly with each other and with the "objective" procedure across sentences. This underlines the applicability and validity of the test in combination with time saving subjective assessment methods. Moreover, the variability in performance across different sentences correlates inversely with the RMS level of the respective sentence. This indicates that an adjustment of sentence material with respect to RMS level already yield reasonably homogeneous test material with respect to intelligibility. PMID- 9348700 TI - Cryptic invasions of the crab Carcinus detected by molecular phylogeography. AB - Coastal marine ecosystems world-wide are threatened by invasions of nonindigenous species. The ubiquity of marine sibling species identifiable only by genetic analysis suggests that many invasions are cryptic and therefore undetected, causing an underestimation of the actual number and impacts of invading species. We test this hypothesis with European crabs in the genus Carcinus that have invaded five regions globally. Partial 16S ribosomal RNA gene sequences confirm sibling species status of morphologically similar Atlantic C. maenas and Mediterranean C. aestuarii. Based on 16S rRNA haplotypes, crabs from California, New England and Tasmania were all C. maenas. However, we report the cryptic multiple invasion of both species in Japan and South Africa, where only C. aestuarii and C. maenas, respectively, were previously recognized. PMID- 9348701 TI - An evaluation of introgression of Atlantic coast striped bass mitochondrial DNA in a Gulf of Mexico population using formalin-preserved museum collections. AB - Striped bass Morone saxatilis populations in drainages along the Gulf of Mexico coast (Gulf) were depleted in the 1950s and 1960s, probably because of anthropogenic influences. It is believed that only the Apalachicola-Chattahoochee Flint (A-C-F) river system continually supported a naturally reproducing population of Gulf lineage. Striped bass juveniles of Atlantic coast (Atlantic) ancestry were introduced to restore population abundances in the A-C-F from the late 1960s to the mid 1970s and in many other Gulf rivers from the 1960s to the present. We previously identified mtDNA polymorphisms that were unique to approximately 60% of striped bass from the A-C-F and which confirmed the continued successful natural reproduction of striped bass of Gulf maternal ancestry within the system. However, the genetic relatedness of the extant A-C-F population to 'pure' Gulf striped bass was not addressed. In this study, we determined the frequency of a diagnostic mtDNA XbaI polymorphism in samples of 'pure' Gulf striped bass that were collected from the A-C-F prior to the introduction of Atlantic fish, that were obtained from museum collections, and that were originally preserved in formalin. PCR primers were developed that allowed for amplification of a 191-bp mtDNA fragment that contained the diagnostic XbaI restriction site. Using RFLP and direct sequence analyses of the PCR amplicons, we found no significant differences in mtDNA XbaI genotype frequencies between the archived samples and extant A-C-F samples collected over a 15-year period. This indicates that significant maternally mediated introgression of Atlantic mtDNA genomes into the A-C-F gene pool has not occurred. Additionally, we found no evidence of the unique Gulf mtDNA genotype in striped bass from extant populations in Texas, Louisiana and the Mississippi River. These results highlight the importance of the A-C-F as a repository of striped bass to restore extirpated Gulf populations and the potential use of museum collections in retrospective population studies. PMID- 9348702 TI - Genetic population structure of the Greater Bilby Macrotis lagotis, a marsupial in decline. AB - The Greater Bilby has shown a rapid decline in range during this century and now occupies only a small isolated area in south-western Queensland (QLD) and a larger, but mostly low-density area in the north-western deserts of the Northern Territory (NT) and Western Australia (WA). We have examined variation in the control region of mitochondrial DNA (mtDNA) and at nine microsatellite loci in order to investigate the extent of current and historical subdivision across the species range, and to provide a preliminary assessment of genetic structuring and mating system on a finer scale within the QLD population. Both mtDNA and microsatellite loci had substantial variation within and among populations, with mtDNA divergence being greater between QLD and NT than between NT and WA. The QLD population had two unique and divergent mtDNA lineages, but there was no evidence for strong phylogeographical structure across the range. The available evidence suggests that the bilby should be considered as a single Evolutionarily Significant Unit consisting of multiple Management Units. Augmentation of the remnant QLD population from the NT does not appear necessary at this stage, at least not on genetic grounds. Finer-scale analysis of microsatellite variation for two QLD colonies revealed a deficiency of heterozygotes and significantly greater relatedness within than between colonies. However, structuring was observed only for males; relatedness values for females did not depart from those expected under panmixia. Parentage exclusion analysis for one colony allowed the construction of a partial pedigree which indicated strong polygyny, with one male fathering all but one of the eight offspring assigned. The extent to which fine scale genetic structuring and differences between sexes is due to sex-biased dispersal vs. effects of mating system remain to be determined. PMID- 9348703 TI - Molecular evolution at Mhc genes in two populations of chinook salmon Oncorhynchus tshawytscha. AB - The DNA sequences of four exons of the MHC (major histocompatibility complex) were examined in chinook salmon (Oncorhynchus tshawytscha) from an interior (Nechako River) and a coastal (Harrison River) population in the Fraser River drainage of British Columbia. Mhc class I A1, A2 and A3 sequences and a class II B1 sequence were obtained by PCR from each of 16-20 salmon from each population. The class I A1 and a pair of linked A2-A3 exons were derived from two different classical salmonid class I genes, Sasa-A and Onmy-UA, respectively. Allelic variation for B1, A1 and A2 was characterized by the high levels of nonsynonymous substitution indicative of the effects of natural selection on Mhc domains that contain peptide binding regions. The number of alleles detected at each of the four exons ranged from three (B1) to 22 (A1), but levels of nucleotide sequence divergence at all four exons were low relative to classical mammalian Mhc genes. The nucleotide similarity among alleles ranged between 89 and 99% over all exons, and all four domains possessed only two major sequence motifs. Allelic distributions at B1, A1 and A3 confirmed the genetic distinctiveness of the Harrison and Nechako chinook salmon populations revealed in previous studies. The two major allelic motifs of B1 and A1 segregated strongly between the populations. In spite of evidence that allelic diversity at these chinook salmon Mhc exons has been generated by selection, the level and distribution of diversity in the two salmon populations strongly reflected the demographic history of the species, which has been characterized by repeated bottlenecks and isolation-by-distance in glacial refugia. PMID- 9348706 TI - NIOSH moving ahead on agenda. PMID- 9348704 TI - Phylogenetic characterization of ineffective Frankia in Alnus glutinosa (L.) Gaertn. nodules from wetland soil inoculants. AB - Ineffective Frankia endophytes were retrieved from various wet soils by using Alnus glutinosa clones as trapping plants. No pure cultures could be isolated from these ineffective nodules. Therefore, the phylogenetic position of these endophytes was determined by sequence analysis of cloned PCR products of bacterial 16S rDNA, derived from nodules. The results showed that all nodule endophytes belong to a hitherto undescribed cluster of the Frankia phylogenetic tree. The position of these uncultured ineffective Frankia nodule endophytes is different from that of the ineffective Frankia isolates derived from A. glutinosa nodules, even when originating from the same geographical location. This suggests a bias in current isolation techniques. PMID- 9348707 TI - Ergonomic modeling with MQPro. PMID- 9348708 TI - The brave New World of PPA. PMID- 9348710 TI - New strategies for hearing protection. PMID- 9348709 TI - A comprehensive safety and health program for the small employer. PMID- 9348711 TI - Better, safer driving. PMID- 9348712 TI - Plantibodies. Human antibodies produced by field crops enter clinical trials. PMID- 9348715 TI - The pulmonary hemodynamic effects of 1,1,1-trichloroethane inhalation. AB - We examined the hemodynamic effects of 1,1,1-trichloroethane (TCE) inhalation in anesthetized dogs in acute inhalation experiments. Six adult mongrel dogs with spontaneous respiration were intubated, connected to a one-way valve. TCE was delivered by the tubular system connected to a 30-liter tedlar bag reservoir filled with 1 v/v% of TCE. After TCE inhalation, the animals revealed a significant decrease in systemic arterial pressure following a decrease in systemic vascular resistance. Pulmonary arterial pressure, pulmonary vascular resistance and right cardiac work increased significantly. These changes were compatible with clinical manifestations of TCE intoxication in human beings. In conclusion, inhalation of TCE may not only decrease peripheral vascular resistance, but also induce transient disturbance of pulmonary blood flow with subsequent pressure overloading in the right ventricle. PMID- 9348716 TI - Direct effect of inorganic mercury on citrate uptake by isolated rat renal brush border membrane vesicles. AB - Occupational exposure to mercury has long been associated with renal proximal injury and an increased incidence of proteinuria, as has such exposure to cadmium. Renal citrate excretion is very important with respect to acid-base balance since the metabolism of citrate generates three bicarbonate ions. In this study, we exposed isolated rat renal brush border membrane vesicles (BBMV) to mercury (Hg2+) and examined their citrate uptake characteristics. BBMV were prepared by the divalent cation precipitation method. Citrate uptake was measured by the Millipore rapid membrane filtration method. The preincubation of BBMV with 0.5 and 2 mM HgCl2 for 1 min significantly inhibited citrate uptake compared with that of BBMV without Hg preincubation. The analysis of the time course of citrate uptake during a 30-min preincubation of BBMV with 0.1 mM Hg2+ also revealed a significant reduction in the uptake compared with that of the control BBMV without preincubation. These findings indicate that the preincubation of BBMV with mercury results in a time- and concentration-dependent inhibition of citrate uptake. PMID- 9348717 TI - Concentration in blood and organs of dogs after high dose 1,1,1-trichloroethane inhalation. AB - Dogs were exposed to 1% (v/v) (10,000 ppm) vapor of 1,1,1-Trichloroethane (1,1,1 TCE) by inhalation for 3 min repeated four times at 4 hr intervals under continuous anesthesia. Changes in the 1,1,1-TCE concentration in blood with time, as well as distribution of 1,1,1-TCE in the organs and tissues (lungs, liver, kidneys, heart, brain, and fat around the kidneys and on the abdominal wall) upon completion of the four exposures were studied. Concentrations of 1,1,1-TCE in blood showed the highest level immediately after exposure, and fairly decreased in about 30 min after exposure. The half life of 1,1,1-TCE in blood was 4-12 min after exposure. Upon completion of the exposures (3 min inhalations repeated four times), 1,1,1-TCE concentrations per gram wet weight of each organ ranged from 0.1 to 0.5 microgram/g in the lungs, liver, kidneys, heart and brain. On the other hand, the 1,1,1-TCE concentration in fat ranged from 16.9 to 54.6 micrograms/g, greatly exceeding those in other organs. PMID- 9348718 TI - Coal workers' pneumoconiosis: a study of prevalence in coal mines of eastern Madhya Pradesh and Orissa states of India. AB - With objective to find out prevalence of Coal Worker's Pneumoconiosis and variation among readers in reading x-ray plates for pneumoconiosis, a retrospective epidemiological survey of Coal Worker's Pneumoconiosis was undertaken in 72 collieries of Madhya Pradesh and Orissa by re-reading of x-ray plates taken during the Periodical Medical Examination at the Occupational Health Units over a period of 5 years. Six readers, trained abroad in reading pneumoconiosis x-ray plates, were involved for the study. Each reader reported approximately one sixth of the available x-ray plates of all the collieries and classified on the 12 point scale of I.L.O. (International Labour Organisation) 1980 in special format. Total 43,504 chest x-rays were reviewed. The overall prevalence was found to be 3.03%, ranging from 1.52% to 4.76% between 10 areas (group of mines). Major category of profusion was category-I (81.09%), followed by category-II (17.84%). Only 3 cases of Progressive Massive Fibrosis (PMF) were detected. Round shaped opacities are predominant (89.59%) in Coal Worker's Pneumoconiosis. Among the opacities, 'p' type is more prevalent (48.29%) followed by 'q' type (40.62%). There was variation amongst the different readers and ranged from 1.14% to 6.76% for reporting the prevalence of Coal Worker's Pneumoconiosis. However, when analysis of six readers for inter reader variation was conducted, that shows no abnormal deviation in the reading of any of the readers. PMID- 9348719 TI - A simple method for carbon disulfide monitoring using a diffusive sampler, thermal desorption and a stain tube. AB - A simple sampling and analytical method for monitoring carbon disulfide (CS2) vapor was investigated to assess exposure to low levels of CS2 in a viscose rayon factory. CS2 vapor was adsorbed on polymer beads (poly (2,6-diphenyl-p-phenylene oxide)) packed in a diffusive sampling tube. The sampling tube was heated at 180 degrees C for 7 min using a Daily Exposure Limit Test Apparatus, and thermally desorbed CS2 was measured by a stain tube for CS2. In laboratory experiments, the indicated CS2 levels measured by this method were highly correlated with calibrated CS2 concentrations (1-40 ppm), exposure duration (1-8 hr) and cumulative exposure levels. The CS2 values were stable up to 7 days after sampling when the diffusive tubes were stored at 4 degrees C and 20 degrees C. The effects of relative humidity, wind velocity and hydrogen sulfide on the measured values were negligible. In a field survey, 65 workers in a viscose rayon factory wore both the diffusive sampling tube and a commercially available 3M 3500 organic vapor monitor on their collar during their 8-hr work period. CS2 concentrations obtained by the two methods were comparable and the correlation coefficient was 0.931. This method proved to be useful in determining the concentrations of CS2 to which workers were exposed. PMID- 9348720 TI - An infrasound experiment system for industrial hygiene. AB - An infrasound experiment system has been constructed in order to investigate the effects of infrasound on human health from the point of view of industrial hygiene and to establish a method to assess it in working environments. Some measurements were carried out to evaluate the acoustic performance of the system in the low frequency sound range, including infrasound, and to clarify the points to be improved. As a result, it was found that the system had some weak points in the audible frequency range. They were considered to be caused by the large capacity of the test chamber, and some countermeasures were required. On the other hand, in the infrasound range, it appeared that the higher harmonics of infrasound possibly affected the experiments and some limitations might be found necessary. But the frequency response of the chamber and the spatial uniformity of the reproduced infrasound proved to be satisfactory for the experiments in this range. Consequently it was concluded that the system could contribute to the study of infrasound. PMID- 9348721 TI - Determination of specific mercapturic acids as an index of exposure to environmental benzene, toluene, and styrene. AB - Methods were developed for the determination of urinary phenylmercapturic acid (PMA), a metabolite specific for benzene, benzylmercapturic acid (BMA), a metabolite of toluene and phenylhydroxyethylmercapturic acids (PHEMAs), specific for styrene, in human beings. Methods involved sample clean up followed by deacetylation and derivatization of the compounds with o-phthaldialdehyde and 2 mercaptoethanol. The fluorescent derivatives were separated on reversed-phase columns with gradient runs and detected by a fluorescence detector. The detection limits were 0.5 microgram/l for PMA and BMA, and 7 micrograms/l for PHEMAs. The background levels of PMA were higher in smokers than in nonsmokers, while no difference was found in the levels of BMA and PHEMAs. Coexposure to ethanol enanched the excretion of BMA in subjects experimentally exposed to toluene. Correlations were found between environmental benzene (r = 0.74, log transformed data), toluene (r = 0.74) or styrene (r = 0.56) and specific mercapturic acids in workers. The usefulness of PMA, BMA and PHEMAs as biomarkers is critically evaluated. PMID- 9348722 TI - Comparative study of challenge aerosols for performance test for dust respirators. AB - A comparative study of challenge aerosols was conducted to review a performance test for dust respirators. The national approval test for dust respirator certification in Japan requires that air containing quartz particles of smaller than 2 microns in diameter should be used as the test aerosol. Aerosols with broad size distributions may therefore be used as the test aerosols. In view of the international harmonization of respirator certification standards, it is necessary to use alternative test aerosols for the approval test for dust respirators. The present study was undertaken to measure the collection efficiency of filters by using three kinds of test aerosols, i.e., quartz dust, sodium chloride and dioctyl sebacate mist aerosols. We used the cartridges of dust respirators and filtering facepieces from eight Japanese and foreign manufacturers, all of which have been certified by the national approval test. Good correlation among the measured collection efficiencies was found for the three test aerosols, but penetrations with sodium chloride and dioctyl sebacate mist aerosols were more than 10 times those of quartz dust aerosol. PMID- 9348723 TI - The characteristics of specific IgG to phthalic anhydride (PA)-albumin conjugate. AB - Phthalic anhydride (PA), used in the chemical industry, binds to proteins and causes allergic reactions. It is important to study the characteristics of antibody to PA-protein. We produced specific IgG against PA-rabbit serum albumin (RSA) by administering subcutaneous injections of PA-RSA conjugate to two rabbits. Both rabbits' sera had high titers of IgG not only to PA-RSA but also to PA-human serum albumin (HSA) and HSA. In enzyme-linked immunosorbent assay (ELISA) and ELISA HSA inhibition, specific IgG to PA-HSA revealed cross reactivity to three other phthalyl anhydride conjugates, hexahydrophthalic anhydride (HHPA)-HSA, methylhexahydrophthalic anhydride (MHHPA)-HSA, and methyltetrahydrophthalic anhydride (MTHPA)-HSA, in both sera. Titers of IgG to HHPA-HSA, MHHPA-HSA, and MTHPA-HSA were not significantly different. On affinity chromatography, highly specific IgG to PA hapten alone was purified. In the serum not binding to PA column, specific IgG to PA-HSA was significantly less than in original serum, but levels of specific IgG to other phthalyl anhydride-HSA were unchanged. Rabbits immunized with PA-RSA produced at least two types of IgG: one is to PA hapten alone and the other may be against new antigenic determinants (NADs) on HSA. PMID- 9348724 TI - The effects of nitroglycol on rat isolated cardiac muscles. AB - To elucidate the action of nitroglycol (Ng) on cardiac muscles, the contractile and chronotropic responses of the isolated rat cardiac muscles to Ng in a cumulative manner were investigated. Ng produced negative chronotropic and inotropic effects on spontaneously beating right atria in concentrations ranging from 10(-7) to 3 x 10(-4) M. Ng also produced dose-dependent negative inotropic effects on electrically driven left atrial muscles. On the other hand, in right ventricle muscles, Ng induced positive inotropic effects. These results suggest that Ng acts directly on the cardiac muscles as well as vascular smooth muscles in acute poisoning. PMID- 9348725 TI - Changes in plasma lipoproteins as toxicity markers for carbon tetrachloride, chloroform, and dichloromethane. AB - Effects of single intraperitoneal (i.p.) administration of carbon tetrachloride (CCl4), chloroform (CHCl3), and dichloromethane (CH2Cl2) on lipoproteins in plasma and liver were investigated in rats. Changes in lipoproteins caused by these solvents were compared with changes in traditional hepatotoxicity markers such as GPT (ALT). Following the administration, concentrations of lipoproteins (VLDL, LDL, HDL), triglyceride, cholesterol, and GPT activity in plasma were determined through changes in liver weight, liver content of triglyceride, malon dialdehyde (MDA), and glutathione (GSH). Time-course study revealed that changes in plasma and liver reached their peaks at 19 or 32 hr following the administration of CCl4 or CHCl3. Peaks of changes were observed at 8 or 19 hr following the administration of CH2Cl2. Dose dependency of these changes was investigated at dosages of 3, 30, and 300 mg/kg of CCl4 or CHCl3, and 300, and 1,000 mg/kg of CH2Cl2. Significant decreases in triglyceride and apolipoproteins in VLDL fraction were observed at 3 mg/kg of CCl4. Such VLDL components decreased at 30 mg/kg of CHCl3. HDL decreased significantly at 300 mg/kg of CH2Cl2 and marked increase in LDL occurred at 1,000 mg/kg of the solvent. Liver weight and liver content of triglyceride and MDA significantly increased at 30 mg/kg of CCl4, while significant increase in GPT activity was observed at 300 mg/kg of CCl4 and CHCl3. GPT increased significantly at 1,000 mg/kg of CH2Cl2. These results revealed that changes in plasma lipoproteins can serve as sensitive and simple markers for liver disorders caused by chlorinated hydrocarbon solvents such as CCl4, CHCl3, or CH2Cl2. PMID- 9348726 TI - Response to psychological test in elderly patients with hand-arm vibration syndrome and healthy controls. AB - We investigated the parasympathetic nervous response to psychological test using heart rate variation (HRV) during deep breathing in elderly patients with hand arm vibration syndrome and healthy controls. Average age (SD) of 16 patients with vibration-induced white finger (VWF), 13 patients without VWF and 12 healthy controls was 60.1 (2.8), 60.6 (4.2) and 58.8 (3.8), respectively. After an initial supine rest for 40 min, psychological test (Stroop color word test and mirror drawing test) was performed for 20 min. The indexes of HRV (Mean R-R, SD, RMSSD and CV) were calculated. Blood pressure and heart rate were also measured. The indexes of HRV did not differ between the groups before exposure. The SD, RMSSD and CV of the patients without VWF during supine deep breathing after 3 min post-exposure supine rest were significantly lower than those of the control group (p < 0.05). The Mean R-R of the patients without VWF significantly increased (p < 0.05). Blood pressure did not differ in either before or after exposure measurements. The results suggest that the post-exposure response of the parasympathetic nervous system to psychological test reduced in the patients without VWF. PMID- 9348727 TI - A study on the relationship between blood lead levels and anemia indicators in workers exposed to low levels of lead. AB - The workers were classified into two groups according to their median value of blood and urine lead levels and urine ALA levels. The group of workers whose blood lead levels were in a lower range (1-15 micrograms/dl) exhibited significantly lower hematocrit, hemoglobin and erythrocytes than the group of the workers whose blood lead levels were in a higher range (16-39 micrograms/dl) (P < 0.001). However, there were no significant differences in the mean values of hematocrit, hemoglobin and erythrocyte counts between the workers whose urine lead or ALA levels were below the median value and those whose urine lead or ALA levels were above the median value. Because the bulk of blood lead is reported to exist in the soluble fraction of erythrocytes, low levels of blood lead are believed to exhibit low levels of anemia indicators. PMID- 9348728 TI - In vivo lipid peroxidation responses of tissues in lead-treated Swiss mice. AB - A single intraperitoneal injection of lead acetate (200 mg/kg body weight) increased the lipid peroxidation potential (LPP) measured as thiobarbituric acid reactive substance (TBA-RS) in different tissues of Swiss mice. All the tissues taken for experimentation, generated significantly higher amount of TBA-RS in lead-treated mice when compared with the respective control value. However, none of the tissues could correspond to the control value after the lapse of four weeks post-treatment. Possibilities of differential responsiveness of tissues to generate lipid peroxides in lead-treated mice have been discussed. PMID- 9348729 TI - Genetic localization of the type I trimethoprim resistance gene and its dissemination in urinary tract isolates in Taiwan. AB - In a total of 425 urinary isolates of E. coli, Enterobacter spp., and Klebsiella spp. selected, there were 169 (45.4%) isolates harbouring type I dihydrofolate reductase (DHFR) gene among 374 trimethoprim-resistant isolates. In these 169 isolates, only 17.2% hybridized with the Tn7 probe. According to another probe specific for the integrase gene of integron, 87.6% showed a positive reaction. Further analysis by restriction mapping proved that the type I DHFR gene was inserted into a integron-like structure. These results indicate that the type I DHFR gene that was initially observed in association with transposable element Tn7 is becoming associated with an integrase function similar to integrons in most instances. Further analysis of the distribution of Tn21-like integrase gene in clinical isolates indicated that the prevalence rates were 86.4%, 84.8%, and 76.7% respectively in E. coli, Enterobacter spp., and Klebsiella spp.. Furthermore, the integrase gene found in our clinical isolates proved to be mediated by a plasmid, demonstrated by Southern hybridization. Thus, the trimethoprim-resistant gene that developed under selective pressure from the double drug trimethoprim and sulphonamide was transmitted by insertion into integron-like structure and then mediated by plasmid transfer for dissemination. PMID- 9348730 TI - Frozen-section diagnosis in surgical pathology: a quality assurance study. AB - In a quality assurance study we reviewed one thousand four hundred and forty three consecutive frozen sections performed at department of pathology, VGH-TC from June 1995 to July 1996. The diagnostic accuracy was 92.6%. The diagnosis was deferred in sixty-eight cases (4.7%). False positive for malignant tumor was made in two cases (0.14%) and false negative diagnosis for malignancy in thirty-seven (2.56%). The inaccurate diagnosis was mainly in samples taken from the brain, female breast, and thyroid. Incorrect diagnoses were mainly due to interpretation of the pathologic findings (71.8%), followed by gross sampling (15.4%) and microscopic sampling (12.8 %). Some of the lesions were difficult to diagnose even in permanent sections. Technical skill and diagnostic expertise are essential for frozen diagnosis. We suggest that an accuracy survey of frozen section be periodically performed in every pathology department as part of its quality assurance program. PMID- 9348731 TI - Intravenous repletion of phosphorus deficiency in the chronic renal failure patients with severe hypophosphatemia. AB - Severe hypophosphatemia is a potentially life-threatening medical condition and might lead to a fatal outcome in critically ill patients. The situation is further complicated by the co-morbid renal failure. We evaluated the efficacy and safety of the intravenous phosphate repletion in 15 renal failure patients with severe hypophosphatemia. Six patients with advanced renal failure and nine patients under maintenance hemodialysis, 7 males and 8 females, aged between 42 and 83 years old, were found to have serum phosphate level < 1.2 mg/dL from various medical conditions and were treated with intravenous phosphate infusion. The phosphate solution prepared from sodium dihydrogen phosphate (NaH2PO4), containing 13 mg/ml phosphate and 0.5 meq/ml sodium, in the dosage 2.5-3.0 mg phosphate/Kg body weight, was administered through the central venous lins every 6-8 hours. The infusion was discontinued once serum phosphate level reached 5.0 5.5 mg/dL. Serum ionized calcium, phosphate and intact parathyroid hormone levels were serially followed at different intervals, respectively. The hemodialyzed uremic patients received their dialysis treatment as scheduled. All patients survived the hypophosphatemic period and regained normal phosphate levels after repletion. The amount of phosphate administered to reach the target level ranged between 3438 and 9150 mg and the duration of treatment varied between six and seventeen days. Hypocalcemia (< 4.2 mg/dL) was noted at eight occasions during the whole treatment period but none was symptomatic. Eleven patients recovered from the offending illness. However, four patients expired due to reasons not directly consequent to and temporally remote from hypophosphatemia. We conclude that prompt repletion of severe hypophosphatemia and phosphate deficiency with relatively slower rate of NaH2PO4 solution intravenous infusion is a safe and effective mode of treatment for renal failure and uremic patients. The longer treatment period allowed the administered minerals full equilibration. The risk of hyperkalemia is avoided and the sodium/volume load can be eliminated by dialysis. PMID- 9348732 TI - Characteristics of adolescent patients and their health problems at an adolescent health clinic. AB - In Taiwan, there has been an increase in biopsychosocial morbidity among adolescents in the past ten years, but neither an adolescent health care delivery system nor a subspecialty training program in adolescent medicine has been hitherto established. To better understand the characteristics of adolescent patients and their health problems, the sociodemography and health problems of all adolescent patients aged 11 to 21 years who visited the Adolescent Health Clinic twice or more during January, 1994 and December, 1995 at a college hospital, were analysed by reviewing patients' medical records and the adolescent registration records. There were totally 264 adolescent patients with an average age of 18.7 +/- 3.4 years. The number of female adolescents involved in the study (139 persons or 52.7%) was little more than that of male adolescents. Most of the adolescent patients (67.4%) were in late stage of adolescence; in addition, most of them (81.4%) were students. During the 2-year study period, there was a total of 350 independent health problems among 957 office visits made by the 264 patients, averaging 1.3 problems and 2.7 office visits per patient and per problem, respectively. Supplementary classification, mental disorders, and respiratory system diseases were the three leading disease classifications, occupying 66.6% of all disease classifications. General medical examination, upper respiratory tract infection, and immunization were more common among all individual diagnoses. Regarding gender difference in the encountered health problems, hepatitis B, office visiting for counseling, and conductive disorder, were more prominent in the male adolescents, whereas upper respiratory tract infection, goiter, acne vulgaris, peptic ulcer disease, migraine, and eating disorders were more common in the female adolescents. Moreover, the patients in the different stages of adolescence had their unique health problems. It was concluded that there were variations in the health problems encountered among adolescents of different genders and of the different stages of adolescence. PMID- 9348733 TI - Surgical treatment of pathologic fracture of the femur. AB - A retrospective study of the surgical treatment of 32 metastatic lesions of the femur in 30 patients at the Kaohsiung Medical College Hospital was performed from 1987 to 1994. There were 16 women and 14 men with an average age of 61 years. A surgical technique combining internal fixation or prosthesis and methylmethacrylate cement was used in all cases. Adequate pain relief was achieved in thirty-one cases (97 %). Of the entire group, 20 cases (62%) remained ambulatory, 10 cases (31%) were confined to a wheelchair, only two cases had implant failure and one suffered from infection. PMID- 9348734 TI - Influence of rigid gas permeable contact lens solutions on corneal epithelial wound healing. AB - To determine the effect of RGP contact lens solution on corneal epithelial wound healing, the following solutions including Soaclens, Contopharma GPHCL-S, Boston condition, Bausch & Lomb condition and Duracare were applied on corneal epithelial wounds of enucleated pig eyes to evaluate possible cytotoxicity of RGP solutions. The wounds, created by excimer laser, were 1.5mm in diameter with 70 microns in depth. The eyeballs were maintained in an incubator using a perfusion system. After twenty-four hours, a score from 3 to 0 was given depending on the size of defect from absence of healing to completely healing. The average scores of the epithelial defect in each group are: Soaclens: 0.38 +/- 0.74; GPHCL-S: 0.63 +/- 0.52; Boston condition: 0.38 +/- 0.52; Bausch & Lomb condition: 0.25 +/- 0.46 and Duracare: 2.38 +/- 0.52. Most of the epithelial wounds healed with one exception, the eyeballs which received Duracare still had large defects. The difference of scores between Duracare and other groups are statistically significant. Duracare, which contains benzalkonium chloride, may be responsible for retarded wound healing. PMID- 9348735 TI - Alteration of glucose uptake in cultured human corneal endothelial cells grown in high glucose media via cAMP-dependent pathway. AB - In this study, cultured human corneal endothelial cells were incubated in media containing various concentrations of glucose at 5 mM, 10 mM, and 25 mM for 2 days. Then, the cellular 2-deoxyglucose uptake and cAMP concentration of cultured human corneal endothelial cells were measured. The results indicated that the activity of cellular glucose uptake of nmole/min/mg protein was decreased gradually from 0.18 (5 mM), 0.10 (10 mM), 0.07 (20 mM) to 0.06 (25 mM) after 2 days incubation with a high concentration of glucose. The glucose uptake in insulin-treated human corneal endothelial cells also exhibited a similar declining effect in high glucose media from 0.30 (5 mM), 0.11 (10 mM), 0.08 (20 mM) to 0.05 (25 mM). The cAMP concentration in human corneal endothelial cells was measured in the presence of high glucose media. It was indicated that the cAMP concentrations of pmole/well in both insulin-treated and non-insulin treated cells were also decreased after increasing the glucose concentration in the media from 73 (5 mM) to 20 (25 mM) and 101 (5 mM) respectively. The cAMP concentration in insulin-treated cells was less than in non-insulin treated cells. This decreasing effect was significantly reversed by the addition of 1 mM dibutyryl cAMP to the cells for 1 hour in both groups. These results suggest that the diabetic state may decrease the 2-deoxyglucose uptake in human corneal endothelial cells via cAMP-dependent pathway. PMID- 9348736 TI - Laparoscopic fenestration for giant liver cyst. AB - Laparoscopic fenestration for treatment of the non-parasitic cyst of the liver has been rarely reported, but sporadic cases appeared elsewhere in the literature. Here we report four cases with symptomatic giant nonparasitic liver cysts which were treated by a laparoscopic fenestration procedure that allowed the successful removal of the cyst dome. Before starting to excise the wall of the cyst, laparoscopic-quided needle aspiration of the cyst fluid was done first in order to clean the visual field for laparoscopic intervention where possible. The cyst wall was usually slightly transparent and somewhat smooth in the external and internal surface of the cysts. It was necessary to lysis the omental adhesion sometime before starting to remove the dome of the cyst. The cyst wall of the exposed part could be removed first with heat-probe instrument through laparoscopy. Those patients were discharged and revealed an uneventful post operative course in three cases but in one case we had to convert to the traditional laparostomy to perform resection of the multiple cystic lesions. Post operative echographic study showed that the giant cyst had collapsed. Therefore, we believe laparoscopic fenestration for the liver cyst is simple and effective, if the patient is a candidate who requires operation to remove the dome of the giant cyst. PMID- 9348737 TI - Epidural abscess presented with psychiatric symptoms. AB - Cranial epidural abscesses are unusual in neurosurgical practice. Mostly they are secondary to skull bone osteomyelitis of foreign body implantation as a result of trauma, or infection of paranasal sinus, otitis, and mastoiditis in adults or late adolescents. The purulent inflammatory process of the epidural abscess, thrombophlebitis of the venous drainage, septic thrombosis, direct extension into the orbit, carvenous sinus, superior orbital fissure give the epidural abscess a high mortality and morbidity. We present an interesting case, who has had psychiatric symptoms such as bizarre behavior, auditory and visual hallucination for about two years. Incidental brain computed tomograms, to exclude the organic somatic disorder, revealed a huge brain abscess. Emergent surgical intervention was carried out and the episodes of talking to himself and auditory hallucination subsided. The removal of the epidural abscess eliminated the symptoms and cured the patient. All the right amygdata, entorhinal area, cingulate gyrus, hippocampus and parahippocampal gyrus of this patient were compressed by the huge abscess. All these structures belonged to limbic system. Diseases involving the limb system may cause emotional disturbances, such as delusions, illusions and hallucinations, emotional lability, pathological laughing and crying, rage reaction and aggression, apathy and placidity, even endogenous fear, anxiety, depression and euphoria. Dramatic improvement of the patient was found after surgical removal of the abscess. We highlight this interesting case for it will undoubtedly bring together a large cooperation of psychiatrists, neurologists and neurosurgeons. PMID- 9348738 TI - Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis--a case report of corneal ulcer. AB - We report a case of corneal ulcer caused by nontuberculous mycobacteria which was confirmed by smear and culture. We attempted a new method for the rapid identification of mycobacteria to the species level on the basis of evaluation by the polymerase chain reaction of the gene encoding. The method is involved with restriction enzyme analysis of PCR product obtained with primers common to all mycobacteria. Using the restriction enzyme Bst EII and Hae III, clinically relevant and other frequent laboratory isolates were differentiated to the species or subspecies level by PCR-restriction enzyme analysis. The main prevalence of pattern analysis is Mycobacterium chelonae subsp. abscessus in this case. The outcome suggests that PCR-restriction enzyme analysis should be a useful method for early diagnosis concerning nontuberculous mycobacterial keratitis. PMID- 9348739 TI - A result on a 2 x 2 survival experiment. AB - Lifetime data classified according to categorical variables under the proportionality of the hazard functions of response variables for various treatment combinations is assumed. The proposed model is a combination of Cox's proportional hazards model and ANOVA model. The existence of a solution to the marginal likelihood function is examined for the case of 2 x 2 two-way classification. We provide an easily verifiable condition for the existence of a unique estimate. PMID- 9348740 TI - Closed-loop stability of pharmacokinetic-pharmacodynamic models. AB - In automatic feedback control of intravenous drug infusions, convergence to the setpoint is an important objective. This paper examines the stability of pharmacokinetic-pharmacodynamic models of patient response regulated with proportional integral feedback. The model consists of three components: linear compartmental pharmacokinetics, a first-order lag, and sigmoidal static pharmacodynamics. The permitted pharmacokinetic models obey the principle of detailed balance and admit drug administration into and sampling from the same compartment. Convergence to the setpoint occurs if the reset time of the controller is greater than the maximum possible time constant of the first-order lag. The convergence analysis uses standard Popov stability theory and takes advantage of the little known fact that many pharmacokinetic models possess poles and zeros that alternate on the negative real axis. PMID- 9348741 TI - A mathematical model of breast and ovarian cancer treated with paclitaxel. AB - A mathematical model that describes the effects of cell-cycle-specific drugs on cancer and normal tissue is developed. The model takes into account the proliferating cells, which are sensitive to the treatment, and the quiescent cells, which are resistant to the treatment. With the use of information from the medical literature, model parameters are estimated for breast and ovarian cancer as well as for bone marrow. Then, with the use of the model and the estimated parameters, some acceptable treatment strategies are discussed in terms of treatment period, drug-infusion time, and proliferative fraction of cancer mass. Finally, these results are compared with current clinical practices for treatment with Taxol, and possible improvements on current treatment strategies are suggested. PMID- 9348742 TI - Mathematical description of synergistic interaction of hyperthermia and ionizing radiation. AB - A simple mathematical model of simultaneous combined action of ionizing radiation and hyperthermia has been proposed. The model suggests that the synergistic interaction of ionizing radiation and hyperthermia is expected to result from the additional lethal damage arising from the interaction of sublesions induced by both agents. These sublesions are considered nonlethal after each agent taken alone. The model was applied to the quantitative analysis of the simultaneous action of hyperthermia and ionizing radiation. It predicts the dependence of synergistic interaction of the ratio of lethal events produced by every agent used, as well as the maximal value of the synergistic effect, conditions at which the maximal interactive effect can be achieved, and the dependence of synergistic effect on dose rate. The predictions of the model have been tested with four experimental data sets reported in the literature. The theory appears to be appropriate and the conclusions valid. PMID- 9348743 TI - No liability for consultation with treating physician. PMID- 9348744 TI - Physician responsibility and accountability. Future legislation may clarify issues. PMID- 9348745 TI - Physician accountability. New home health care law increases penalties. PMID- 9348747 TI - The Banbury Conference. Genomics to physiology and beyond: how do we get there? PMID- 9348746 TI - Certified medical assistants. Health care's versatile professionals. PMID- 9348748 TI - Successful aging. An overview. PMID- 9348749 TI - Ways to make "usual" and "successful" aging synonymous. Preventive gerontology. AB - Preventive gerontology is the study and practice of those elements of lifestyle, environment, and health care management that will provide the maximal longevity of highest quality for individuals and the population. As such, it focuses on a personalized hygiene agenda that varies in its emphasis according to a person's age, sex, and risk factor profile. It includes a matrix of strategies relating to diet, exercise, and the avoidance of substance abuse and adverse environmental exposure. Preventive gerontology carries differential emphases according to the life stage of a person, featuring long-term, low-cost, and low-risk lifestyle strategies in youth and middle age (generally to age 75) and more short-term, low risk interventions in old age (> 75), especially secondary prevention, according to individualized estimates of risk, cost, and benefit. The aggregate effect of widespread application of this approach--especially insofar as it is coupled with a rising level of education and continued psychosocial development--will be progressive congruency between usual and successful aging. A by-product will also be an ever-advancing median age of the population and, inevitably, a growth in long-term health and social service needs. Responsible planning for this consequence of success in the 21st century will require a rededication of North Americans to care for those in need regardless of age. PMID- 9348750 TI - Building communities that promote successful aging. AB - Despite the fact that, in a few years, a fifth of the US population will be older than 65 years and people will be living a third of their lives after retirement, we have developed few avenues that would permit older adults to play meaningful roles as they age and few institutions to harness the experience that older adults could contribute to society. In fact, older adults constitute this country's only increasing natural resource--and the least used one. In this article we consider the rationale for developing institutions that harness the abilities and time of older adults, rather than focusing solely on their needs. Such an approach would decrease the structural lag between a social concept of retirement as unproductive leisure and an aging population that is larger, healthier, and with a need for more productive opportunities. Gerontologically designed opportunities for contribution on a large social scale could well provide a national approach to primary prevention to maintain health and function in older adults. PMID- 9348751 TI - Healthy aging. A women's issue. AB - The life expectancy of women currently exceeds that of men by almost seven years, yet women spend approximately twice as many years disabled prior to death as their male counterparts. The diseases that account for death and health care utilization in older women (heart disease, cancer, stroke, fracture, pneumonia, osteoarthritis, cataracts) are also major contributors to disability. This paper reviews the scientific evidence that supports specific recommendations for older women that may prevent or delay these conditions for as long as possible. Risk factors for falls and fractures should be assessed and, where possible, modified. Adequate intakes of calcium, vitamin D, fruits, and vegetables should be encouraged. Weight should be monitored and weight loss discouraged for most women. Screening for B12 deficiency is recommended. Engaging women in a shared decision-making process about the use of hormone replacement therapy for longterm prevention of heart disease and fractures is important, as is regular screening for breast and colo-rectal cancer. Women should be encouraged to engage in enjoyable physical activities, including walking, for 30 minutes daily. These interventions have the potential to delay the onset and improve the course of many chronic conditions that prevail in later life. PMID- 9348752 TI - Optimal medication use in elders. Key to successful aging. AB - Pharmacotherapy represents one of the most important ways in which the practice of geriatric medicine differs from conventional medical care. The older patients is a major consumer of prescription and nonprescription medications, and proper use of these agents can lead to more cost-effective strategies in reaching optimal health. A key difference in distinguishing appropriate from inappropriate drug use is evident in the themes of polymedicine and polypharmacy. Polymedicine describes the use of medications for an older population for the treatment of multiple co-morbid conditions, while polypharmacy represents a less-than desirable state with duplicative medications, drug-to-drug interactions, and inadequate attention to pharmacokinetic and pharmacodynamic principles. The purpose of this paper is to outline strategies toward optimal medication use as a key to successful aging. Specifically, we discuss themes of cost-effective prescribing, the role of medication compliance, overuse and underuse of medication, over-the-counter products, alcohol abuse, and preventive medicine. In addition, we discuss policy implications and responsibility for ensuring the high quality of pharmaceutical care. The reader should have a practical understanding of the pertinent issues in geriatric clinical pharmacology and its relationship to successful aging. PMID- 9348753 TI - Successful aging calls for more than drugs. PMID- 9348755 TI - Hear ye? Hear ye! Successful auditory aging. AB - Age-related hearing loss (presbycusis) is a multifactorial process that affects nearly all people in their senior years. Most cases are due to a loss of cochlear hair cell function and are well mediated by communication courtesy and modern amplification technology. Severe hearing loss is generally due to cochlear problems or age-related diseases and may require speech reading, assistive listening devices, and cochlear implants, depending on the degree of loss. Presbycusis may seriously impair communication and contribute to isolation, depression, and possibly dementia. Accurate diagnosis and prompt remediation are widely available but are frequently underused. Geriatric health care and well being is enhanced by the detection and remediation of communication disorders. PMID- 9348754 TI - Heart health in older adults. Import of heart disease and opportunities for maintaining cardiac health. AB - Coronary heart disease remains the leading cause of morbidity and mortality in older adults, despite improved survival and declining mortality. This article describes the prevalence and impact of heart disease on people's lives, singly and in combination with other diseases. It then reviews current findings as to the risk factors for CHD in older adults and the underlying physiologic changes of aging plus pathophysiologic changes of hypertension and CHD in impairing the ability of older adults to respond to exercise and other stressors, and the effects of exercise training in attenuating the adverse cardiovascular changes of aging. This information provides a basis for considering opportunities for prevention of heart disease and maximizing heart function. The article concludes by describing the known contribution of preventive measures to declines in heart disease in older adults. PMID- 9348756 TI - Seeing into the future. Vision and aging. AB - The leading causes of visual impairment in North Americans are age-related, but appropriate care can preserve useful vision for most older adults. Cataract surgery is highly successful. Early detection and treatment of glaucoma can prevent vision loss. Laser treatment is remarkably effective against diabetic retinopathy. Vision loss due to macular degeneration cannot be delayed in all patients, but low-vision rehabilitation can maximize the usefulness of remaining sight. PMID- 9348758 TI - Overcoming barriers to successful aging. Self-management of osteoarthritis. PMID- 9348757 TI - Preserving mobility in older adults. AB - Age-related loss of strength contributes to impaired mobility and increases the risk of falls. Recent research has focused on 2 approaches to preventing age related loss of strength--promoting physical activity and exercise (especially strength training) and using trophic factors to enhance muscle performance. Epidemiologic evidence strongly supports a role of regular physical activity in successful aging by preserving muscle performance, promoting mobility, and reducing fall risk. Randomized controlled trials provide convincing evidence that strength and endurance training improve muscle performance in older adults. Evidence is rapidly accumulating from randomized trials that endurance, strength, and balance training promote mobility and reduce fall risk, though exercise effects differ according to the type of exercise, details of the exercise program, and the target group of older adults. Because lifetime regular physical activity is recommended for all older adults, a reasonable strategy (especially for weak adults) is an activity program that includes strength training. In contrast, insufficient evidence exists to recommend the long-term use of trophic factors to preserve muscular performance. An intervention that merits additional study is avoiding the use of psychoactive drugs because drugs like benzodiazepines appear to be risk factors for inactivity and may have unrecognized direct effects on muscular performance. Because chronic illness is a risk factor for inactivity and disuse muscle atrophy, randomized trials comparing strength training with other interventions would be useful in understanding whether strength training has advantages in preserving muscle performance and improving health-related quality of life in a variety of chronic illnesses such as depressive illness. PMID- 9348759 TI - Memory, thinking, and aging. What we know about what we know. AB - Cognition is the foundation that underlies all daily activities, from the most basic to the most complex. Successful aging depends, in large part, on maintaining a level of cognitive ability that allows a person to interact effectively and appropriately with the environment. In the following article we provide an overview of the effects of aging on cognition; discuss physical, social, and psychological factors that have been shown to influence cognition in old age; and review current literature on interventions that may optimize successful cognitive aging. We conclude with a discussion of abnormal cognitive aging and review current research on risk factors and treatments of Alzheimer's disease and other dementing illnesses. PMID- 9348760 TI - Maintaining good morale in old age. AB - Traditional aging studies have seen life's later years as a time of inevitable biological and social decline. Psychological decline might also be expected, but this is not true for most older people, according to epidemiologic studies. Thus, we must ask: Why is aging not as emotionally threatening as might be expected? Why do some older people do better than others? How should medicine address these issues? It is only possible to understand the successful emotional aging of most elders if the customary diathesis-stress model is supplemented by a developmental perspective. Expectations as well as capacities diminish with aging. This means that subjective health appears more tightly linked with morale than objective health. Some older people experience recurrence of mental disorders (for example, major depression) first present earlier in life. Others experience new disorders such as minor depression in response to biological or social losses. As geriatric medicine comes to increasingly focus on chronic disease, attention to morale is an important strategy for maximizing quality of life. Physicians will need improved skills in the detection and treatment of problems in morale if they are to provide optimum care for their older patients. PMID- 9348761 TI - Aging and sexuality. AB - Recent research suggesting that a high proportion of men and women remain sexually active well into later life refutes the prevailing myth that aging and sexual dysfunction are inexorably linked. Age-related physiological changes do not render a meaningful sexual relationship impossible or even necessarily difficult. In men, greater physical stimulation is required to attain and maintain erections, and orgasms are less intense. In women, menopause terminates fertility and produces changes stemming from estrogen deficiency. The extent to which aging affects sexual function depends largely on psychological, pharmacological, and illness-related factors. In this article I review the physiological sex-related changes that occur as part of the normal aging process in men and women. I also summarize the effects on sexual function of age-related psychological issues, illness factors, and medication use. An understanding of the sexual changes that accompany normal aging may help physicians give patients realistic and encouraging advice on sexuality. Although it is important that older men and women not fall into the psychosocial trap of expecting (or worse, trying to force) the kind and degree of sexual response characteristic of their youth, it is equally as important that they not fall prey to the negative folklore according to which decreased physical intimacy is an inevitable consequence of the passage of time. PMID- 9348762 TI - Community resources for frail older patients. AB - The goal of community-based services for frail older patients is to help them achieve the greatest degree of functional ability and independence. The services available include case management, geriatric assessment, adult day health care, home health services, and the Program for All-inclusive Care for the Elderly (PACE). Definitive criteria for referral have not been established, but without some targeting, the efficacy of these services remains uncertain. Targeting criteria identified include dependency in 2 or more activities of daily living, no family support, dementia, many long-term illnesses, and many hospital stays. Although efficacy and cost-effectiveness remain uncertain, patients, families, and physicians generally report these services to be helpful. PMID- 9348763 TI - Preventing decline in function. Evidence from randomized trials around the world. AB - Over the past decade, increasing epidemiologic evidence points to a few important risk factors for loss of mobility and functional decline, such as inactivity, cigarette smoking, obesity, depression, and alcohol and drug misuse. Global interventions addressing these risk factors along with other medical problems have been evaluated in randomized controlled clinical trials. With few exceptions, these trials have shown that intervention has a positive effect on health and functional status, as well as reducing length of stay in a hospital or nursing home. The extent to which the success of these programs is attributable to risk-factor reduction or to geriatric assessment and management is uncertain. PMID- 9348764 TI - Culture, universals, and the personal. AB - This chapter summarizes a part of the case that can be made that the individual construction of a personal domain of choice and privacy generalizes across cultures and is not restricted to persons who live within Western or so-called modern societies. The research findings reported here are consistent with the view that persons seek to establish such areas of control in order to maintain a differentiated personal identity and a sense of personal agency. Children, adolescents, and adults from the United States and traditional cultures have been found to identify a class of behaviors and issues as being outside the legitimate sphere of social or moral regulation. Mothers from Western and traditional cultural settings recognize and foster their children's claims to areas of personal choice and privacy. Across cultures, as children mature and move toward adulthood, they lay claim to a broader range of issues and actions as personal matters. Research on adolescent-parent conflict with U.S. and Chinese samples has indicated that these shifts associated with adolescent claims to freedom are the source of most family conflicts. Anthropological accounts of adolescent-parent conflicts in 160 cultures have provided evidence that such conflicts are widespread (Schlegel and Barry, 1991). Finally, we are beginning to obtain evidence that parental overcontrol of personal issues is associated with symptoms of psychological problems in their adolescent children. These research findings are consistent with the proposal (Nucci, 1996) that establishment of a personal domain is an intrinsic feature of normal human development, resulting from the inevitable attempt by individuals to account for and differentiate between their own motives, values, and experiences and those of others. The evidence also points to the fact that such personal issues are coexistent with concerns for interpersonal harmony and social integration. Thus, it is not surprising that the work summarized here also suggests considerable social-class and cultural variation in how the personal is expressed. Such variation is consistent with the assumption that the personal is constructed out of social interactions (Nucci, 1996) that entail reciprocal interchange between individual and societal structures (Turiel, 1996). In Spiro's analysis (1984), the results of such reciprocal structural interaction cannot be accounted for by reducing the analysis of psychological structures in terms of cultural structures and vice versa. Thus, any accurate interpretation of the impact of culture on psychological development must be constrained by features that are peculiar to psychological systems. Extending this to the cross-cultural study of the personal domain, a case can be made for the need to explore such issues at the level of the individual, rather than at the level of the cultural shared-symbol system. On the other hand, this nonreductionist approach and the available evidence rule out the reification of the personal as a culturally empty set of psychological issues. As illustrated in studies of the distribution of rights in relation to gender and social hierarchy among Druze Arabs (Wainryb and Turiel, 1994), the interplay between the personal and the cultural system of roles and obligations provides a rich and contradictory portrait that can be understood only by shifting perspective from the social to the individual and back again without favor. PMID- 9348765 TI - Bayous and jungle rivers: cross-cultural perspectives on children's environmental moral reasoning. PMID- 9348766 TI - Moral heteronomy in context: interviewer influence in New York City and Recife, Brazil. PMID- 9348767 TI - Negotiating the methods debate: context is the real issue. PMID- 9348768 TI - Caring for the dying: nurses' experiences in hospice care. AB - This article reports on a phenomenological study of nurses' experiences of caring for dying patients in a Western Australian hospice. Data obtained from indepth interviews with nine experienced hospice nurses were analysed using Colaizzi's method. The five major themes that emerged from the data indicated that: nurses were transformed by the experience of caring for dying patients; the hospice context influenced caring; caring was embodied in nurse-patient interactions; caring extended to patients' families and nurses developed strategies to cope with their experience. The implications of the findings are that awareness of the effects of their caring activities on patients, their families and themselves is essential to nurses' maintenance of self and to their ability to enable patients to make choices. Suggestions are made for future research of the importance of caring contexts in both comforting patients and in preparing them for death. PMID- 9348769 TI - The body of the nurse as patient. AB - This small qualitative study was an exploration of how female nurses had viewed their bodies when they were patients in acute care hospitals. Analysis of data obtained in individual interviews with six nurses revealed that they had perceived themselves to be inferior, disciplined and disembodied and that they had protected their privacy intensely. The implications of the findings and recommendations for further inquiries are discussed. PMID- 9348770 TI - Self-care: re-thinking the role of compliance. AB - This paper argues for a research approach that recognises that people make choices about their compliance to medical regimens according to their own perceptions of their needs, regardless of whether these conform to professionals' perceptions of needs. This approach requires an acceptance of a construction of reality that allows for personal experience to be accepted as having a truth of its own. The relationship between a reconstructed conceptualisation of self-care and the emancipation and empowerment of people with chronic illnesses is discussed. PMID- 9348771 TI - The effect of a nursing intervention on the incidence of older patient falls. AB - This paper describes the evaluation of a fall prevention protocol that combined the assessment of the mobility and confusion status of 2,023 patients aged 70 years and over and a toileting regimen for at risk patients who were both confused and having mobility problems. The six months' study was conducted in a 450 bed metropolitan teaching hospital and involved approximately 500 nursing staff in the hospital's medical and surgical wards. Almost five percent (4.7%) of patients in the study group fell; 13 patients fell more than once and the total number of falls was 112. Twenty-four percent of patients (n = 482) were assessed as being at risk of falling and 54% of falls (n = 61) occurred in the at risk group. Sixteen percent of these falls occurred in the sub-group who had been toileted according to the study protocol and 84% in the sub-group who had not been toileted according to the protocol. There were 53% fewer patient falls during shifts which complied with both the assessment and toileting protocol than during non-compliant shifts. Given the simplicity and effectiveness of the study protocol, the finding that it was not followed on 43% of shifts is of concern. PMID- 9348773 TI - Core curricula, certification, and holistic nurses. PMID- 9348772 TI - Nurses facilitating reconciliation through education. AB - In 1996, Health of Indigenous Australian Peoples, was introduced as a compulsory unit in a Bachelor of Health Science in Nursing course and as an elective unit for students from other courses. The first cohort of students in the unit were surveyed to determine whether their attitudes to 21 issues that affect the health of Aboriginal people were more appropriate at the end of the unit than they had been before the unit. The findings were that the student group's post-unit attitudes were more appropriate in all 21 areas. The results support a recommendation that units designed to increase health workers understanding of issues that affect the health of Aboriginal people must be compulsory if reconciliation is to be achieved between Aboriginal and non-Aboriginal people. PMID- 9348774 TI - A focus on education. PMID- 9348775 TI - Natural cycles. PMID- 9348777 TI - Commune-ication as sacred circle. PMID- 9348776 TI - American Holistic Nurses' Association, revision of bylaws. PMID- 9348778 TI - The uniqueness of nursing. PMID- 9348779 TI - Natural cycles. PMID- 9348780 TI - Insites: your computer link. Electronic teacher. PMID- 9348781 TI - Holistic nurse entrepreneur--growing a business. PMID- 9348783 TI - Spiritual healing. PMID- 9348782 TI - The Monitoring and Actualization of Noetic TRAining the MANTRA Study, a randomized controlled study of healing interventions. PMID- 9348784 TI - Holistic nurse of the year: a transformational journey. PMID- 9348785 TI - The continuing evolution of the certification process. PMID- 9348786 TI - Explaining the accreditation process. PMID- 9348788 TI - Natural cycles. PMID- 9348787 TI - In-sites: your computer link. "The little engines that could: what's new in searches"? PMID- 9348789 TI - Millie Freel named 1997 holistic nurse of the year. PMID- 9348790 TI - Earning of contact hours. PMID- 9348791 TI - The uniqueness of nursing. PMID- 9348792 TI - Natural cycles. PMID- 9348793 TI - Postpartum homecare. PMID- 9348794 TI - Legal status for direct entry midwives in your state. PMID- 9348795 TI - Interview with Gert Welsh, CNM. Interview by Ina May Gaskin. PMID- 9348796 TI - When peer review is not enough. PMID- 9348797 TI - Gift of life. PMID- 9348798 TI - Common vision, common goals: keynote to the fifth annual Birth Gazette Conference. PMID- 9348799 TI - Inspiring women: the fifth annual Birth Gazette Conference. PMID- 9348800 TI - Maternal mortality in the United States: where are the doctors? PMID- 9348801 TI - Toxic legacy--the poisoning of America's children. PMID- 9348802 TI - Looking for the fingerprint of a teratogen. PMID- 9348803 TI - Gauging the health of school health. PMID- 9348804 TI - Listen and hear, reader urges. PMID- 9348805 TI - Merging the CNS and NP roles. PMID- 9348806 TI - More thought needed before any merger. PMID- 9348807 TI - Section 43 should not be repealed. PMID- 9348808 TI - Dietitians on trans-fat in breast milk. PMID- 9348810 TI - Outpost nursing: a legend comes alive. PMID- 9348809 TI - Kingston Conference links breast cancer and environment. PMID- 9348811 TI - The cycle of life. An experience with qualitative data analysis. PMID- 9348812 TI - Daring to color outside the lines. PMID- 9348813 TI - Osteoporosis. Preventing and treating an insidious disease. PMID- 9348814 TI - Managing postoperative gas pain. PMID- 9348815 TI - Delegation and liability. PMID- 9348816 TI - Living through a revolution. PMID- 9348817 TI - Discovering cultural importances. PMID- 9348818 TI - Understanding ethnic women's experiences with pharmacopeia. AB - For centuries, women have been the primary caregivers and healers in their communities. The literature documents well that the tasks involved in caregiving, such as preparing remedies, providing physical nurturance, and attending the sick, give a gynocentric flavor to healing traditions. Ethnic women, in particular, promote caring and curing through a wide range of folk remedies. These remedies employ foods, herbs, and over-the-counter medications. Importantly, the actual remedies are not as critical to the care of an ill person as the meaning of the cultural memories inherent in acts of caring. I discovered comfort, nurturance, and familiarity to be intrinsic to the holistic nature of the remedies used and to be salient features of the memories of healing and curing within Puerto Rican and African American groups. Through the use of remedies, the Puerto Rican and African American women who participated in these phenomenological investigations impart healing for the body, mind, and spirit. PMID- 9348819 TI - Women's images of midlife: observations from the Seattle Midlife Women's Health Study. AB - Our purpose in conducting this study was to determine how a cohort of women born between 1935 and 1955 defined midlife, and what midlife events they viewed as important, distressing, and satisfying. A random sample of women enrolled in the Seattle Midlife Women's Health Study (n = 131) participated in a telephone interview about the meaning of midlife and important events occurring during the past year. They described midlife similarly to women from earlier birth cohorts with one important exception: the centrality of work and personal achievements in their lives. Contemporary midlife women's views of midlife reflect their roles in society. PMID- 9348820 TI - Women's patterns of contraceptive use. AB - Our purpose in conducting this research was to examine women's patterns of contraceptive use. Records for 800 women receiving care at a private, nonprofit agency providing well-woman and contraceptive care over a 15-year period constitute the data set. The records were examined for patterns of reported method choice and use, method destinations, first method choice, and demographic variables. Women reported making 1,889 method choices from among 16 different methods. Seventy-five percent of the women changed methods at least once and the women gave 1,036 reasons for changing methods. Oral contraceptives and condoms were the methods most commonly tried by the women. The women's patterns of contraceptive use were very individualized, were neither linear nor predictive as other investigators have reported, and method destinations could not be predicted from previous method use. PMID- 9348821 TI - Lactation and the labor market: breastfeeding, labor market changes, and public policy in the United States. AB - Public health authorities in the United States actively promote breast-feeding, with target goals for increased beast-feeding rates by the year 2000. In recent decades, however, there has been an increase in the number of American mothers with infants who are in the labor market. Drawing together research examining the intersection of breast-feeding and women's involvement in paid employment, as well as various labor market analyses, this study explores how national recommendations advocating increased breast-feeding among new mothers in paid work are reconciled with economic pressures to return to the labor force in the early postpartum period. This analysis highlights those employment-related factors that constrain the practice of breast-feeding, thereby impeding "choice" over infant feeding method for many mothers. Finally, there is an attempt to explore various employer and public policies and strategies potentially supportive of breast-feeding among mothers in paid employment. PMID- 9348822 TI - New directions in the treatment of men who batter women. AB - Singular paradigms and simple solutions are not sufficient in addressing the complex and historically sanctioned practice of wife battering. In this article I examine two philosophical approaches to the treatment of men who beat women and how epistemology shapes the assumptions that drive interventions and guide research. A selected review of 8 years of outcome research in batterer intervention reveals inconsistencies in measuring both physical and psychological violence and in how and when those measures are obtained. In addition, there has been a failure to examine violence in the context of community that includes court-ordered treatment and probation monitoring. Failure to address the epistemological assumptions behind why men beat women results in poorly developed interventions and may endanger women as well. PMID- 9348823 TI - The internal structure of the Eating Disorder Inventory. AB - The Eating Disorder Inventory (EDI; Garner, Olmsted, & Polivy, 1983) is used as a screening instrument for identifying potential anorexia nervosa and bulimia nervosa. It comprises eight subscales: Bulimia, Drive for Thinness, Body Dissatisfaction, Maturity Fears, Introceptive Awareness, Perfectionism, Interpersonal Distrust, and Ineffectiveness. We examined the internal structure of the EDI using a principle components factor analysis, for a sample of women with no known eating disorder. The results of the factor analyses showed differences between the extracted factors and Garner et al.'s subscales. Seven factors were extracted, accounting for 60.8% of the variance. The first factor accounted for 33.5% of the variance. The analysis indicated that items factored along positive-negative and factual-emotional lines. We conclude that at present there is not enough evidence to support the efficacy of the subscale structure with women who are not known to have eating disorders. PMID- 9348824 TI - [Violence: abuse--consequences for pregnancy. Interview by Arne Gravanes]. PMID- 9348825 TI - ["Communism protects against allergies". Interview by Eldri H. Fagerlund]. PMID- 9348826 TI - [Vaccines: time-consuming for conscientious parents. Interview by Ingrid M. Hoie]. PMID- 9348827 TI - [Youth and sexuality: healthy or risky life style?]. PMID- 9348828 TI - [Midwifery practice without insurance reimbursement. Interview by Eldri H. Fagerlund]. PMID- 9348829 TI - [Work of mourning: memory box for stillborn infant]. PMID- 9348831 TI - Using standardized patients and standardized physicians to improve patient-care quality: results of a pilot study. AB - BACKGROUND: We conducted evaluation research with a sample of registered professional staff nurses in a large, inner-city, tertiary medical center for a pilot study of videotaped case scenarios using standardized patients and standardized physicians to enhance nurses' communication and collaboration skills. METHOD: Change scores from pre-test to post-test on a self-reported rating scale to assess nurse-physician-patient interactions and communications for 28 nurses were compared with a control group of 38 nurses who did not participate in the videotaped sessions. RESULTS: Repeated measures of analysis of variance (ANOVA) detected no statistically significant differences between the intervention and control groups. However, positive changes were noted in some aspects of nurse-physician and nurse-patient interactions in the intervention group. Immediate feedback from the videotaped scenarios heightened nurses' awareness of the impact of their body language. CONCLUSIONS: Nurses must continuously practice and enhance their collaborative and communication skills. This pilot study suggests that it is beneficial to use videotaping with standardized patients and standardized physicians to enhance such nurses' skills. PMID- 9348830 TI - Time and change. PMID- 9348832 TI - Application of the critical pathway and integrated case teaching method to nursing orientation. AB - BACKGROUND: Nursing staff development programs must be responsive to current changes in healthcare. New nursing staff must be prepared to manage continuous change and to function competently in clinical practice. METHOD: The orientation pathway, based on a case management model, is used as a structure for the orientation phase of staff development. The integrated case is incorporated as a teaching strategy in orientation. The integrated case method is based on discussion and analysis of patient situations with emphasis on role modeling and integration of theory and skill. RESULTS: The orientation pathway and integrated case teaching method provide a useful framework for orientation of new staff. Educators, preceptors and orientees find the structure provided by the orientation pathway very useful. CONCLUSION: Orientation that is developed, implemented and evaluated based on a case management model with the use of an orientation pathway and incorporation of an integrated case teaching method provides a standardized structure for orientation of new staff. This approach is designed for the adult learner, promotes conceptual reasoning, and encourages the social and contextual basis for continued learning. PMID- 9348833 TI - Identification, intervention and education: essential curriculum components for chemical dependency in nurses. AB - BACKGROUND: A documented need exists for continuing education in the area of chemical dependency as it relates not only to patient care, but also to nurses who are susceptible to addiction. This is significant due to the fact that nurses are at risk for chemical dependency and many nurse peers are unable to recognize the signs of chemical dependency and therefore unable to actively intervene. CONCLUSION: According to the literature, which includes current research, nurses lack knowledge regarding specific risk factors, symptoms of chemical dependency in peers, and steps for intervention. In addition, the literature revealed that nursing curricula allot little time to chemical dependency issues. The results of a small-scale learning needs assessment support this literature finding. Continuing education courses can effectively educate nurses to be able to identify their own susceptibility and those of chemically dependent peers, intervene appropriately, and begin the healing process for the impaired nurse. This article outlines a curriculum and additional resources to address the learning needs of nurses related to chemical dependency. PMID- 9348834 TI - Substance abuse education for clinical nurses: a controlled study. AB - BACKGROUND: A study was conducted to evaluate a substance abuse component of a workshop for nurses being promoted to the position of Advanced Clinical Nurse. METHODS: We compared whether the 88 nurses who received the educational intervention increased their knowledge and enhanced their feelings of competence regarding the care of chemically dependent patients more than a control group of nurses who received the promotion workshop without the substance abuse component. RESULTS: The nurses in the intervention group had greater increases in knowledge and competence. CONCLUSIONS: This study points out the importance of providing hospital nurses with continuing education on substance abuse to compensate for their educational deficiencies, to provide the information they need and desire, and to help them meet the ANA's practice standards. PMID- 9348835 TI - Student pre-entry experience and first year of employment. AB - BACKGROUND: Student clinical and work-related experiences are available at many healthcare organizations, but little is known about how these experiences contribute to the employer/nurse work relationship that begins after graduation. This study examined the relationship of senior BSN students' past employer experience in describing first-year employer commitment and turnover. METHODS: The sample was 63 senior BSN students. Experience factors measured included job selection factors, pre- and one-year commitment, organizational climate, employer support, and first-year turnover. RESULTS: Organizational climate was the experience-related factor most important in explaining first-year commitment. CONCLUSION: Findings suggest the importance of supportive work environments to new nurses both before and after accepting the first graduate nurse position. PMID- 9348836 TI - Gaming: a teaching strategy to enhance adult learning. AB - BACKGROUND: As a nurse educator, I encountered many complaints from staff nurses about mandatory inservice education programs, stating that they are repetitious, time-consuming, often too basic, and at times, downright boring. One exception was an Infection Control Week education session that was done in the form of a game. This session set attendance records and had very positive feedback from staff nurses. As a result of this feedback, the use of gaming as a teaching strategy in nursing education was explored. METHOD: A review of the literature on gaming as a teaching strategy was conducted with special attention to its history, current use, and successes in nursing education. RESULTS: Introduced as a formal teaching strategy more than 75 years ago, gaming offers many advantages over more traditional teaching methods. Games connect theory more closely to real life situation and add innovation, diversity, and the opportunity for immediate feedback. Although gaining in popularity, gaming is not extensively used in nursing education as it is not considered a serious educational tool. However, recent literature suggests much success with its use. CONCLUSION: Gaming as a teaching strategy has proven to be an effective way of conveying information in a stimulating, appealing manner. Games facilitate both beginning and experienced nurses' learning by providing an opportunity for experience without the danger or fear of jeopardizing patient safety. PMID- 9348837 TI - Bathroom blitz it! AB - BACKGROUND: Staff development specialists are always looking for new and better ways to get the JCAHO message to their organization's staff. METHOD: The staff development department used a novel method, the Bathroom Blitz, to educate captive audiences, one-person-at-a-time, thereby using the common situation in an uncommon way. RESULTS: Twelve hundred employees had the opportunity to regularly become informed of JCAHO topics without having to leave the unit. CONCLUSION: The Bathroom Blitz approach was an effective tool for educating staff in preparation for the many "mock surveys" conducted prior to Joint Commission and the actual onsite JCAHO visit. PMID- 9348838 TI - Doctors, nurses and physician-assisted suicide. PMID- 9348839 TI - Identifying training objectives: the role of negotiation. AB - Current training initiatives for health care professionals tend to be based on management assumptions of need or on the wish-list demands of potential course attenders. Each approach has major disadvantages, in that neither has the capacity to simultaneously satisfy the organization's objectives, whilst maintaining the motivation and commitment of the participants. This short report discusses an alternative approach that used a valid and reliable training needs analysis instrument with nurses and managers to inform Nurse Practitioner development. The results facilitated the commissioning of relevant education courses, clarified new role boundaries and acted as an agent for negotiation and change between the two groups. PMID- 9348840 TI - Project to develop an autonomous practitioner programme. AB - The early 1990s has seen a rapid expansion in the nature of clinical roles in health care in response to a number of changes. Throughout 1996, The Oxford Radcliffe Hospital National Health Service (NHS) Trust, in partnership with Oxford Brookes University, School of Health Care, and the Royal College of Nursing (UK) have been collaborating to develop an educational programme at degree and masters level, to support new and evolving clinical roles in acute and primary care, across most professional health registration areas and clinical specialities. To ensure the proposed programme was 'fit for purpose'the Trust commissioned a Training Needs Analysis by the University of Birmingham and a Role Analysis by The Royal College of Nursing. Nurses and Midwives with expertise in practice, management, education and research, have collaborated to develop an innovative educational programme which enables a wide range of public and private health care professionals across acute and community settings to develop their autonomous practice. The programme is fully supported by development of policies which are appropriate to service delivery, support professional practice, multi professional relationships, risk management and patient safety. The issue of contractual and professional responsibilities related to career development have been debated and a proactive strategy agreed. To ensure there is thorough evaluation of the area of practice, research projects to evaluate outcomes for patients as well as practitioners have been designed. This will additionally promote a culture of evidence-based practice. PMID- 9348841 TI - Leadership in British nursing: a historical dimension. AB - A historical overview of nurse leadership in the late 19th and late 20th centuries is presented, supported by relevant material from the literature. The 19th century material revealed the following main themes: emphasis on practical and domestic aspects of management; prominent input of religious ideals and social conscience and, autocratic and feminized style of leadership. The main themes in the contemporary literature examined were: role models in history, dysfunctional leadership styles, importance of knowledge, gender as an influencing factor on nurse leadership and threats to the autonomy of nurse leaders. It was concluded that formal nurse professionalization has progressed steadily during the past hundred years with associated evolution of nurse leaders to fit in with contemporary needs. It is hoped that future policies for nursing will encourage decision-making nearer the 'bed-side', more resource-driven care and value-based leadership. PMID- 9348842 TI - A measure of organizational effectiveness in nursing management in relation to transactional and transformational leadership: a study in a Swedish county hospital. AB - This paper presents an empirical study of the influences of transactional (TA) and transformational (TF) leadership on organizational effectiveness (OE), measured as the degree of goal attainment and the quality of nursing care (NQ). The study subjects were all head-nurses and assistant head-nurses at a medium sized hospital in Sweden (n = 23). The methods used were questionnaires and interviews. The multi-leadership questionnaire earlier developed by Bass was modified and named the Leadership Nursing-Effectiveness Questionnaire (LNEQ), comprising 84 items using Likert-type scales. The study showed low mean scores on OE (2.19) and TA (1.05) but high mean scores on NQ (3.17) and TF (3.84). The results suggest that the degree of TA and TF leadership had a low and insignificant connection with OE in this hospital organization. The study did not support the statement that organizational units exposed to a higher degree of TA and TF leadership at the same time show a high degree of OE, as has been shown in studies in other cultural contexts and organizations. PMID- 9348843 TI - The Buckinghamshire nursing record audit tool: a unique approach to documentation. AB - A project to examine the quality of manual nursing documentation using an action research approach, prior to the introduction of a computerized system is described. The findings showed that there was a general lack of understanding about care planning. There was therefore a need to find an audit tool which could identify and develop nurses' knowledge of documentation as well as identifying the strengths and weaknesses of existing documentation. A literature review revealed Phaneuf's audit tool, but this proved difficult to use. Two nurse teachers agreed to develop their own documentation audit tool, based on the UKCC document on record keeping, which identifies criteria for effective documentation. A tool was developed which adopts a facilitative educational approach enabling the practitioner to audit and learn simultaneously. Much interest is being shown by other areas, and the questions are worded so that it could be used in a variety of settings. PMID- 9348844 TI - Teachers of nursing--presenting a new model of practice. AB - In this paper a new model of practice for nurse teachers is presented. In this model the teacher is identified as an advanced practitioner of nursing but, as an advanced practitioner who specializes in education-in all contexts and across all environments where education and training are taking place. It is also a model of practice-based education which is negotiated, balanced and neither dominated by service or by education, it can be seen as responding to the demands of service and higher education, but not dictated by them. It emphasizes the importance of clinical practice as an essential part of the role of the teacher because, as nurses we are all involved in the practice of nursing. The paper begins by exploring the main issues that impinge upon and continue to affect the development of the teachers role. It then examines the teacher's new role in the practice area and an academic specialist and, finally, looks at the structures and conditions that will be necessary to support this role. PMID- 9348845 TI - Director of nursing and quality: a gilt-edged bond for the Trust? PMID- 9348846 TI - [Concepts and development in health promotion]. PMID- 9348847 TI - [Development of an instrument for adolescent health-promoting behavior]. PMID- 9348848 TI - [A study on nursing and health promotion for menopausal women. Development and evaluation of systematic style modification support at a menopause clinic]. PMID- 9348849 TI - [Concepts of health promotion in nutrition science]. PMID- 9348850 TI - [Report of health promotion activity in Bangladesh as members of JOCV]. PMID- 9348852 TI - [A study of children's experiences with congenital heart disease: using Leininger's ethnonursing method]. PMID- 9348851 TI - [Gender and developmental differences in exercise beliefs among youth, and prediction of their exercise behavior]. PMID- 9348854 TI - [Survey and research (9). Steps in the survey research process for nursing practice: statistics (3)]. PMID- 9348853 TI - [An aid in interpersonal relations and nursing]. PMID- 9348856 TI - MRI brain changes. PMID- 9348857 TI - Left-handed midwives. PMID- 9348855 TI - Tomorrow's health workforce. PMID- 9348858 TI - Maternity rights advice. PMID- 9348859 TI - Shoulder dystocia: diagnosis, prediction and risk factors. PMID- 9348860 TI - Achieving continuity of care and carer. AB - The aims of a new service need to be clarified and agreed to avoid disappointment both for midwives and the women they are caring for. Midwives have different views, expectations and priorities, which may be determined by their previous experience. Providing continuity of care to mixed caseloads is complicated to organise, as women with complications need more time and different skills. The success of any service that aims to provide women with care from a small group of midwives depends on adequate staffing and resourcing. PMID- 9348861 TI - Compulsory caesarean: the ruling of the Court of Appeal. PMID- 9348863 TI - Reading research papers. PMID- 9348862 TI - Smoking cessation programme: an ethical analysis. AB - Many midwives feel it is their moral duty to advise and help pregnant smokers to stop smoking Do midwives have the right to persuade or coerce pregnant smokers to stop smoking? Giving information is not sufficient to help clients to make informed choice Smokers need help to acquire necessary skills to raise self esteem and assertiveness to make informed decisions Many women use smoking as a coping strategy to ensure their own mental well-being and that of their families The majority of women smokers are among the lower socio-economic groups It is essential for midwives to develop strategies that are sensitive and pertinent to the smokers' psychological and socio-economic needs Is it ethical or practical to enforce a total non-smoking policy in a maternity unit? As individuals, pregnant smokers have a 'right' to smoke Pregnant smokers smoke more on admission to hospital Illicit smoking will subject non-smokers to passive smoking and increase incidence of fire hazards It is essential to provide a well ventilated smoking room, thereby respecting the rights of both smokers and non-smokers. PMID- 9348864 TI - The Child Bereavement Trust. PMID- 9348865 TI - Improving management of anemia using CQI techniques. AB - During the last two years, the National Kidney Foundation's Dialysis Outcomes Quality Initiative (NKF-DOQI) Anemia Work Group has been developing clinical practice guidelines for the management of anemia in patients with chronic renal failure. After an in-depth critical analysis of the scientific literature, with a consensus of "expert opinion" used to support recommendations in areas with inadequate published data, evidence-based guidelines were developed. These clinical practice guidelines, regardless of how well-founded in scientific evidence, are likely to be disregarded if they are not applicable in the day-to day care of patients. Thus, they must be practical, clearly stated, and usable, given available resources and constraints. The purpose of this article is to describe our experiences in implementing some of these guidelines over the last two years, as they were being developed, for the care of patients in our hemodialysis unit. PMID- 9348866 TI - Anemia management: one patient at a time. PMID- 9348867 TI - HCFA. Number of dialysis patients reaches over 214,000. Use of living-unrelated kidney donors continues to rise. PMID- 9348868 TI - The Canadian Organ Replacement Register on renal replacement therapy. AB - This report describes the demography, clinical characteristics, and outcomes for patients receiving renal replacement therapy (RRT) in Canada. Results are based on registered patients initiating RRT during the 1981-85 period using data obtained from the Canadian Organ Replacement Register (CORR). PMID- 9348869 TI - "Guts to glory"/Part II. Taking the idea of a new dialysis facility to its shiny reality. PMID- 9348870 TI - Starting from scratch. What makes the ultimate dialysis facility? Wife of ESRD patient asked, and then helped to build it. PMID- 9348871 TI - 1997 U.S. Renal Data System Report. Incidence, prevalence rate showing signs of dropping in ESRD population. PMID- 9348872 TI - DOQI Guidelines/fifth in a series. A focus on iron management for better HcTs. PMID- 9348873 TI - Are we easing up on standard precautions at the wrong time? PMID- 9348874 TI - Hoping for a brighter future. PMID- 9348875 TI - Disability to ability/Part II. Getting prepared for a bodybuilding contest requires an exercise regimen. PMID- 9348876 TI - Attitudes toward care may help implementation of DOQI guidelines. PMID- 9348877 TI - After the tears: surviving a nursing student's suicide. PMID- 9348878 TI - Integrating service with academic study: implementing cultural change. PMID- 9348879 TI - Faculty and student confidence in their clinical judgment. PMID- 9348880 TI - What if? What else? What then? A critical thinking game. PMID- 9348881 TI - A case management curricular model. The challenge for nursing education. AB - Preparing students for practice in a managed care system requires restructuring of the nursing curriculum. The authors describe a case management curricular model as a framework for baccalaureate education. Processes used for curricular redesign and recommendations for knowledge and skills expected of baccalaureate program graduates are included. Examples of expected student case management performance behaviors and evaluation criteria are provided. PMID- 9348882 TI - Integrating theory and esthetics in a graduate nursing theory course. PMID- 9348883 TI - Dealing with difficult students in the classroom. AB - Despite the proliferation of literature on dealing with difficult people, little has been written on how to manage problematic students in the classroom. Every teacher encounters difficult students because the classroom is merely a microcosm of the outside world. Nursing educators strive to create an open and caring atmosphere in their classes, but it is still necessary to intervene when students disrupt the class by showing disrespect for others or by getting the discussion off track. The authors describe the most common difficult student roles seen in the classroom and suggest strategies for dealing with them. PMID- 9348884 TI - Evaluating critical thinking in clinical practice. AB - Although much has been written about measurement instruments for evaluating critical thinking in nursing, this article describes clinical evaluation strategies for critical thinking. Five methods are discussed: 1) observation of students in practice; 2) questions for critical thinking, including Socratic questioning; 3) conferences; 4) problem-solving strategies; and 5) written assignments. These methods provide a means of evaluating students' critical thinking within the context of clinical practice. PMID- 9348885 TI - Standardized measures of critical thinking. Experience with the California Critical Thinking Tests. AB - Standardized measures of student critical thinking are an attractive option for nursing educators under pressure to demonstrate student higher order thinking skills. One program's experience using the California Critical Thinking Skills Test and the California Critical Thinking Dispositions Inventory illustrates some of the problems of using standardized test and potential solutions. PMID- 9348886 TI - Stresses reported by second year nursing students. PMID- 9348887 TI - A perioperative practicum. AB - It is increasingly difficult for nurse educators to provide clinical experiences for students in settings that offer excellent educational opportunities. Hospital administrators are phasing out entry-level medical-surgical nursing positions and decreasing the number of inpatient beds, correspondingly decreasing the numbers of patients available to students. Educators need to develop new strategies to meet this challenge. Perioperative practicums are one method to help accomplish this. The author describes a unique perioperative experience that includes an important home care component. PMID- 9348888 TI - Facilitating multicultural education in maternity nursing. PMID- 9348889 TI - Strategies for getting students on the information superhighway. AB - Many nurses indicate that they lack the formal education needed to use resources such as the Internet and computer technology related to healthcare. The results of a survey at a baccalaureate school of nursing revealed that although nearly 100% of those surveyed used computers, very few effectively used available Internet resources. In response, the authors implemented strategies to make available Internet resources "user-friendly" for communication and information gathering. PMID- 9348890 TI - Health teaching in the community by ADN students. PMID- 9348891 TI - Information processing styles. One size doesn't fit all. AB - By knowing how students process information, educators can get them to notice what is being taught so that they learn as much as possible. Teachers can use this information to change their lessons from lecture and note-taking instruction to active involvement that enhances students' comprehension. The purpose of this article is to examine two major styles of information processing, to identify global and analytic preferences and teaching strategies, and to illustrate how these strategies were applied in a baccalaureate nursing course. PMID- 9348892 TI - Nurses need to be more aware of how much power we have. PMID- 9348893 TI - Suffering and ethical caring: incompatible entities. AB - Ethical problems are continuing to expand in health care due to conflicts of technology and value. This study investigated what kind of ethical problems nurses face in clinical situations and what process they use in deciding on actions to take. Ethical theories in justice and caring were explored. Qualitative research was used and ethnographic analysis was conducted with six staff nurses from three clinical areas. An analysis of the data yielded an overarching theme of 'Suffering and ethical caring: incompatible entities'. Six domains were identified: informant definitions, preceding conditions, actions taken, intervening variables, risks and recommendations. Future research is needed in the field of client and family suffering and pain. PMID- 9348894 TI - Extending the theory of awareness contexts by examining the ethical issues faced by nurses in terminal care. AB - The breaking of bad news in a hospital setting, particularly to patients in a terminal condition, highlights some complex and often emotive ethical issues for nurses. One theory that examines the way in which individuals react to bad news such as a terminal illness, is the theory of awareness contexts. However, this theory may be limited by failing to recognize the complexity of the situation and the ethical issues involved for nurses caring for terminally ill patients. Furthermore, contexts of awareness are influenced to a much greater extent by relationships with nurses than simply by the delivery of medical information. Even when information is given to the client and the family, the nurse is involved in helping them to know the meaning of this information. In a hospital, the nurse is faced with emotional demands by clients, families and colleagues, complex issues of advocacy, and professional boundaries and responsibilities. It is the author's wish to examine the reality of clinical practice for nurses, thus further extending the theory of awareness contexts. PMID- 9348895 TI - Moral orientation of elderly persons: considering ethical dilemmas in health care. AB - Knowledge about moral development and elderly persons is very limited. A hermeneutical interpretative study was conducted with healthy elderly persons (n = 20) in order to explore and describe their moral orientation based on the paradigms of justice (Kohlberg) and care (Gilligan). The types of moral reasoning, dominance, alignment and orientation were determined. All but one participant included both types of reasoning when discussing an ethical conflict. None of the men's moral reasoning was dominated by caring, but justice dominated the reasoning of four women. The implications for ethical decision-making and future research are discussed. PMID- 9348896 TI - Ethical issues experienced by HIV-infected African-American women. AB - The epidemic of human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) has led to many ethical problems. Most studies have focused on the ethical issues faced by nurses who provide care to persons with AIDS (PWA), rather than the ethical issues faced by PWAs themselves. The purpose of this study, therefore, was to explore the ethical issues faced by five HIV/AIDS infected African-American women. An analysis of interview data revealed that these women deal with four broad categories of ethical issues: diagnosis; disclosure; treatment by, and of, others; and future pregnancies. The results of this study provide an initial description of the ethical issues faced by HIV/AIDS infected African-American women, and begin to lay the foundation necessary for nurses appropriately to facilitate and support their decisions. PMID- 9348897 TI - Confidentiality: a critique of the traditional view. AB - 'Confidentiality' can become a somewhat embellishing signboard for paternalistic caring. In essence, one needs to distinguish between confidentiality as a respectful attitude to a patient/client, where it becomes credible that the caring professional will not misuse the information he or she obtains about the patient/client, and between confidentiality misused as an instrument of power to keep the patient/client outside of processes in which it might be important or advantageous for him or her to participate. PMID- 9348898 TI - Self-image, self-values and interpersonal values among newly graduated nurses. AB - This longitudinal study (1994-1996) used the Gordon Personality Inventory to measure nursing students' self-image (Gordon A), self-values (Gordon B) and interpersonal values (Gordon C). It was performed with students from three colleges of health in the south of Sweden: Jonkoping (n = 54), Vaxjo (n = 24) and Kristianstad (n = 38). The null hypothesis of the study was that the new academic three-year programme did not have the power to change significantly the students' self-image and professional values. The hypothesis was tested by paired sample Student's t-test. The result was that, at Jonkoping, self-image changed and increased significantly in the dimensions of 'cautiousness' and 'personal relations', and decreased in 'sociability', and increased in the self-value 'order'. At Vaxjo the self-image dimensions of 'original thinking' and 'personal relations' increased, and, at Kristianstad, the students increased their self image scores in 'responsibility'. PMID- 9348899 TI - Nursing registration in New Zealand was, and is, for life. PMID- 9348900 TI - Lymphoedema: a study of Otago women treated for breast cancer. AB - Lymphoedema, the swollen arm that can follow treatment for breast cancer, is the build up of lymph fluid which is unable to flow normally due to the surgical removal of axillary lymph nodes or the scarring of these nodes from radiotherapy. Previous studies indicate a lack of recognition of preventative measures and treatment of lymphoedema amongst health professionals and women treated for breast cancer. There also appears to be a lack of acknowledgement of the effects that lymphoedema can have on those who develop it. Of 181 women surveyed 68 (38%) reported having developed arm swelling at some stage since their treatment, 56 (31%) of whom met the study criteria for lymphoedema. Of those with lymphoedema 21 (37%) women had not consulted anyone regarding their swollen arm; a further 10 (18%) sought advice, but were offered no treatment. Therefore, in total, 31 (55%) of the women with lymphoedema received no treatment. The remaining 25 (45%) were offered a variety of treatments. In the group of women with lymphoedema, most arm swelling occurred within the first year. For a smaller number, swelling occurred up to 25 years later. Twenty-one percent of those affected, recalled advice about its prevention, compared with 36% in the group who had not experienced lymphoedema. Lifestyles were affected in many ways, with 21 (37.5%) of the women reporting pain of varying intensity and frequency. Other effects were on dress/choice of clothing, household duties, sleep, employment and sports. PMID- 9348901 TI - Learning to be a nurse. The contribution of the hidden curriculum in the clinical setting. PMID- 9348902 TI - Accompanying: the practice of public health nursing in New Zealand. PMID- 9348903 TI - Reading nurses practice as social text: from critical metatherories to personal theories in action. PMID- 9348904 TI - Publishing workshops number 4. Preparing a manuscript: reviewing literature. AB - In this paper we have looked at the different avenues for nurses to contribute to the knowledge base of the discipline by writing about nursing literature. We emphasised the responsibility that goes with this action. As with all writing the starting point is for the author to define the boundaries of the work, and then decide what it is that she or he wants to communicate. After that it is a matter of planning a logical and coherent sequence and proceeding to develop the content in language that is as simple and effective as possible. Reference was made to the general guidelines for writing proposed in an earlier workshop (Chick et al, 1996). In this paper three different forms, a review article, a literature review and review of a piece of literature have been discussed, noting the purposes that each serves and the way in which, to a large extent, purpose dictates both content and the order and priority to be given to the various sections. Three of the key dimensions along which variation can occur are scope, critical emphasis, and relevance to practice. PMID- 9348905 TI - An evidence based nursing centre for New Zealand: in collaboration with the Joanna Briggs Institute for Evidence Based Nursing. PMID- 9348907 TI - Are nurses prepared for PREP? PMID- 9348906 TI - The perennial row over public sector pay. PMID- 9348908 TI - Changing minds. PMID- 9348909 TI - Health of a nation. PMID- 9348910 TI - Testing times. PMID- 9348911 TI - All aspects of nursing are part of the pay agenda. PMID- 9348912 TI - Finding a voice. PMID- 9348914 TI - Breastfeed without pain. PMID- 9348913 TI - An open secret. PMID- 9348915 TI - Public health initiatives in community nursing. PMID- 9348916 TI - Clinical effectiveness and evidence-based practice. AB - In the first of two articles, the authors address some of the most common questions asked by practitioners about clinical effectiveness. The second article, on clinical guidelines, will appear in Nursing Standard in two weeks. PMID- 9348917 TI - Will critical pathways replace the nursing process? AB - Individualised care is the essence of nursing and regard for the individual is the battle against the impersonal institution. Unfortunately, attempts to manage the process of individualised care have often ended in failure (Ford and Walsh 1994). Some observers maintain that the rigid behaviour which characterises the nursing process contradicts the intuitive nature and individualistic approaches which are fundamental to nursing practice (Benner 1984). This article reviews the concept of critical pathways, a new approach to managing patient care. PMID- 9348918 TI - Hypnotherapy: complementary support in cancer care. AB - The psychological and physical consequences of cancer threaten patients' wellbeing and quality of life (Fallowfield 1991). Patients' needs are wide ranging and can include both personal and physical demands as well as support, relaxation and distraction. This article describes how many of these needs can be cared for by the skillful use of hypnotherapy. PMID- 9348919 TI - Caring for patients with cataract. PMID- 9348920 TI - The campaign for devolution. PMID- 9348921 TI - New way of leading. PMID- 9348922 TI - How to speak out and keep your job. PMID- 9348923 TI - When I'm sixty five. PMID- 9348924 TI - 10 years of nursing service. PMID- 9348925 TI - Stress in the community: teaching relaxation. AB - In this article, the author describes a project designed to offer an alternative to traditional treatment for stress. The author discusses the techniques taught to clients to help them cope with stress and the evaluation of the service for users. PMID- 9348926 TI - Cervical screening. PMID- 9348927 TI - Nutrition and wound healing. AB - This paper reviews evidence that suggests malnutrition exists among the sick population in hospital and at home, a situation that has not improved in 20 years. It examines the consequences of malnutrition in broad terms and its effect on wound healing. The role of specific nutrients for wound healing is discussed, and finally the barriers which prevent nurses from delivering adequate patient nutrition are exposed. PMID- 9348928 TI - An alternating pressure trolley mattress in A&E. PMID- 9348929 TI - Nursing and the Internet. PMID- 9348930 TI - An army of helpers. PMID- 9348931 TI - An end to the lottery. PMID- 9348932 TI - Where's the beef? PMID- 9348933 TI - Remembering Diana. PMID- 9348936 TI - Time is of the essence. PMID- 9348934 TI - Coping with aging. PMID- 9348937 TI - Direct line to home. PMID- 9348938 TI - Out of Africa. PMID- 9348939 TI - Handwashing. PMID- 9348940 TI - High time for a change. PMID- 9348941 TI - Mental health--a blast from the past. PMID- 9348942 TI - Dogged pursuer. Interview by Janet Snell. PMID- 9348943 TI - Hot potatoes--cold comfort. PMID- 9348944 TI - Gene therapy for oncology patients. PMID- 9348945 TI - How to achieve effective pain management. AB - Pain is a common problem in a wide range of conditions encountered by nurses in every setting. Despite the availability of knowledge about effective pharmacological and non-pharmacological approaches to pain management, patients still experience uncontrolled pain. This article, the first of six examining how pain control can be made more effective, explores how a basic misunderstanding of approaches to pain relief can lead to ineffective care. The author suggests that improved education and cooperation among professionals can lead to improved pain management. PMID- 9348946 TI - Professional support for HIV and maternity services. PMID- 9348947 TI - Preparing for specialist community nursing practice. PMID- 9348948 TI - Career development opportunities for practice nurses. PMID- 9348949 TI - Planning a career in community nursing. PMID- 9348950 TI - Infection control--screen saver. PMID- 9348951 TI - Infection control--risky business. PMID- 9348952 TI - Know how--i.v. insertion sites. AB - Patients receiving intravenous therapy are at risk of developing an infection at the insertion site. Careful monitoring of the length of time a catheter is in situ, and ensuring that only sterile dressings are used and that they do not allow accumulation of moisture at the insertion site, will reduce the rate of complications. PMID- 9348953 TI - Infection control--washing instructions. PMID- 9348954 TI - Infection control--the situation in hand. PMID- 9348956 TI - A need for global learning. PMID- 9348955 TI - Caring for children. PMID- 9348957 TI - Mental health services for young people: 'inadequate and patchy'. PMID- 9348958 TI - Recovering from burns: Jamie's story. PMID- 9348959 TI - Teenage quality circles ... not just a paper exercise. PMID- 9348960 TI - Transmission of HIV from mother to child. PMID- 9348961 TI - A model for parent education. PMID- 9348962 TI - Childhood cancer: helping the individual to cope within the family. PMID- 9348964 TI - Methicillin-resistant Staphylococcus aureus (MRSA). PMID- 9348963 TI - Management of burns. PMID- 9348965 TI - Self-monitoring of blood pressure in pregnancy. PMID- 9348966 TI - Croup. AB - As many as 3% of children under six years of age are affected by croup, usually at two to three years. Symptoms include a barking cough and inspiratory stridor. The preceding infection of the larynx is usually viral; bacterial infection can complicate the condition. Mist inhalation has been the traditional treatment. Dexamethasone and now budesonide may be used as first-line treatment. PMID- 9348967 TI - Injectable contraceptives: underused and undervalued? AB - Injectable progestogen-only contraceptives can be considered for the woman who is unwilling or unable to use oral contraceptives or an IUD. They have a very low failure rate. They appear to have few serious life-threatening side-effects. The woman does not have to remember to take a daily pill. The method requires little compliance from the client and is independent of patient error. Short-term uses include for partners of men undergoing vasectomy, women being immunised against rubella and for women awaiting sterilisation. Noristerat can be used immediately after an abortion or birth of a baby. Breast feeding is not inhibited. Main side effects are menstrual irregularities and delayed return of fertility after use. It is essential that women are counselled about the method and its side-effects before injectable contraceptives are given. PMID- 9348968 TI - Replacement surfactant therapy. AB - The addition of replacement surfactant therapy to current neonatal intensive care techniques has brought about a significant reduction in the severity of, and mortality from, moderate-to-severe RDS in premature infants. Although exogenous surfactant is relatively easy to administer, its use should be limited to those skilled in handling critically-ill infants on ventilators. While its introduction has been a major step forward, it is not the whole answer to the vexing problem of respiratory distress syndrome. PMID- 9348969 TI - Oral care during pregnancy. AB - There is no reason why pregnancy should result in the mother losing calcium from her own teeth. Many pregnant women find their gums bleed during toothbrushing. This is due to hormonal changes and increased sensitivity to dental plaque. Careful, thorough toothbushing twice a day will help minimise this problem. Sugary snacks are best avoided, but if not it will help preserve dental health to follow them straightaway with foods that stimulate the flow of saliva, such as cheese or sugar-free chewing gum. All pregnant women should be encouraged to attend a dentist early in pregnancy. All NHS dental check-ups and treatment are free during pregnancy and for 12 months after the birth. PMID- 9348970 TI - Advice on medications for coughs and colds. PMID- 9348971 TI - The child, his family and dyspraxia. AB - Children with dyspraxia have difficulties with co-ordination, eg poor balance or problems doing up buttons or writing with a pencil. The cause is unknown. Though mentally normal and with no known neurological condition, these children are unable to plan, organise and co-ordinate their movements. School work is affected and occupational therapy and physiotherapy may be needed. A supportive atmosphere helps counter poor self-esteem. As this study shows, the condition puts strains on the parents and the whole family. Support from the health visitor and school nurse would often be appreciated. With practice, children with dyspraxia can achieve reasonable degrees of motor skills. Children do not necessarily grow out of the condition but can be helped with empathy and appropriate teaching and therapy. PMID- 9348972 TI - Condoms: still the most popular contraceptive. AB - Condoms can be used as a barrier contraceptive and/or to protect against many sexually-transmitted diseases. They are easy to buy and use and free from medical risk. Carefully used, and used in conjunction with a spermicide, condoms have similar reliability to IUDs, progesterone-only pills and the diaphragm. The condom must be put on before the penis touches the vaginal area. The penis should not touch the vaginal area after the condom has been taken off. Oil-based products, eg baby oil, massage oil, lipstick, petroleum jelly, suntan oil, can damage the condom. If a lubricant is required, use one that is water-based. PMID- 9348973 TI - The professional renewal program: aiding nurses in career change. AB - A professional renewal program gives nurses the opportunity to reflect on their current work situation and imagine the opportunities for change and growth in the nursing profession. Nurses return to work refreshed and re-energized, directly affecting their working environment. This article gives specific strategies for developing a professional renewal program. PMID- 9348974 TI - Developing new student clinical sites: a response to downsizing of hospitals. AB - The settings for nursing practice are changing rapidly. Dwindling in-patient numbers and increasing out-patient care have compelled nursing programs to find student "clinical" learning experiences that reflect the changing health care delivery system. This article presents specific strategies for changing the clinical learning practicum experiences in nursing programs to respond to the changes in the health care delivery system. PMID- 9348975 TI - Strategies for a smooth hospital closure. AB - When a hospital closes it becomes the nurse executive's and human resource director's responsibility to ensure the quality of patient care through the transition as well as manage the staff during a stressful time. How do you keep enough staff to care for the patients during the closing period? This author, who has previous experience managing a hospital closure, describes strategies to keep staff and promote patient continuity during the closure process. She emphasizes the need to focus on both patients and staff. PMID- 9348977 TI - Hospital's insurer pays $1.4 million & sues nurses' insurer. PMID- 9348976 TI - When does a nurse become a "supervisor" under federal law? AB - With restructuring and skill-mix changes, nurses are delegating and coordinating nursing skills. Does this mean they are "supervisors" and not protected by the National Labor Relations Act? Not necessarily. These authors clarify several myths related to the Supreme Court ruling on nurses as supervisors and analyze case examples where nurses have been, and have not been, considered supervisors. PMID- 9348978 TI - "Sonogram stat" order ignored: fetus dead--mother dies. Case on point: District of Columbia v. Perez 694 A 2d. 882--DC (1997). PMID- 9348980 TI - Anti-strikebreaker law does not apply to "no strike" situation. Case on point: Prof. Staff Nurses Assn. v. Dimensions Health Corp. 695 A. 2d 158--Md (1997). PMID- 9348979 TI - LA: home health nurse in fatal accident: hospital employer not vicariously liable. NJ: arbitrator awards nurse's aide 275,000 dollars: failure to file timely demand for trial fatal. PMID- 9348981 TI - HIV legislation puts AMA and ANA at odds. PMID- 9348982 TI - Acute care decisions--ethics in action. PMID- 9348983 TI - Lab reports--only as good as the sample. PMID- 9348984 TI - Lab reports--clarifying the CBC. PMID- 9348985 TI - Simple strategies that improve compliance. PMID- 9348986 TI - Childhood trauma--when to suspect abuse. PMID- 9348988 TI - Career options. A cutting-edge job? Consider research. PMID- 9348987 TI - Lipid-lowering agents. PMID- 9348989 TI - Alternative therapies help the dying "live". PMID- 9348990 TI - Patient care management by the primary nurse: a managed care strategy. PMID- 9348991 TI - Activity-based cost management. PMID- 9348992 TI - One payer's perception of managed care. PMID- 9348993 TI - Understanding managed care: past, present, and future. AB - There is no doubt that the continuing complexities of a changing health care system will challenge our competencies and skills. Despite this, those who prosper will maintain a high level of learning, commitment, flexibility, and tenacity. It is a tremendous time to grow managerially in our leadership ability. PMID- 9348994 TI - Managing case management across the continuum: an organized response to managed care. AB - Case management is a core strategy for providing high-quality, cost-effective care in a managed care environment. Acute care case management programs most often originate as nursing-driven, hospital-based initiatives. Given the overwhelming trend toward vertical integration of health care organizations, hospital-based case management programs may be only an interim step toward a longer term strategy. Hospital and case management program leadership should consider how existing case management models can expand to meet patient and provider needs in an integrated health system or network environment. Key issues and strategies for designing a case management system based on continuum of care are described. PMID- 9348995 TI - Role and function within managed care organizations: opportunities for nurse managers. AB - Health care systems are facing rapid changes, moving from a fee-for-service environment to managed care. These changes are impacting nursing, eliminating current jobs and creating new opportunities in the field of managed care. Nurses must take a critical look at their current positions and determine what their contributions will be to improving the system of care. It is imperative that nurses move to leadership positions in managed care to represent the needs of patients and providers at the boardroom table. The purpose of this article is to give an insider's view of traditional and nontraditional roles for nurses within managed care organizations, and to challenge nurses to seize the opportunity to participate in designing the health care delivery system of the future. PMID- 9348996 TI - Management of nursing within a collaborative physician group practice. AB - Today's changing health care environment suggests the need for integration of high-quality primary medical and nursing care to address the complex psychosocial and environmental aspects of health in the elderly. This article describes a model of geriatric primary collaborative care between nurses and physicians that has been successfully implemented in a large primary care group practice setting. PMID- 9348997 TI - Creating a provider network: fact, fantasy, and future. AB - Integrated delivery systems should consider multiple options through which to affiliate, with primary care physicians and advanced practice nurses. Caution should be employed to assure that system alignment occurs in an efficient, effective manner. PMID- 9348998 TI - Managing quality from the utilization management perspective: the provider view. AB - As a contracted provider for a popular health maintenance organization, you receive notice that it is converting its provider contracts to capitated reimbursement models. Reluctantly, you read to see how this change affects your practice. Are you positioned to accept "risk" for your patients? How will you manage your risk? Can you afford to terminate your relationship with this managed care organization and lose your patients to providers who are eager to expand their practice and patient base? This article presents an overview of capitation and ways to manage capitation while promoting quality care and outcomes for your patients. PMID- 9348999 TI - Managing the marketing function for advanced nurse practitioners in a managed care environment. AB - Delivering quality, cost-efficient health care is a desired service in the health care market today. Advanced nurse practitioners are positioned to deliver this product. The key in today's market is clearly defining the product, identifying the customers of the product, and crafting the message for each customer. The development of marketing strategies to address each of the above points will assist an organization in targeting resources and evaluating the effectiveness of the message being delivered. PMID- 9349000 TI - Provider credentialing--managing the quagmire. AB - Credentialing of health care providers by managed care organizations has increased rapidly in recent years. Credentialing as a process is not new, especially to physicians who have for many years been credentialed by hospitals and other institutions. What is new, is going through the process with every individual third-party payor. It is also new, and sometimes mysterious, to advanced practice nurses and other nonphysican providers who are attempting to be included in provider networks for the first time. The key to an effective process is to have efficient, centralized systems in place that manage and track it, and providers and staff who understand the system. PMID- 9349001 TI - [Rehabilitation in brain injuries--the objective is a reorganization of brain function]. PMID- 9349002 TI - [Rehabilitation in brain injury--therapies begin with nursing]. PMID- 9349003 TI - [Rehabilitation in brain injury--a human life in spite of helplessness]. PMID- 9349004 TI - [Rehabilitation in brain injury--the young are especially vulnerable]. PMID- 9349005 TI - [Rehabilitation in brain injury--sociopolitical strategies or health policy management]. PMID- 9349006 TI - [Have fun!]. PMID- 9349007 TI - [One-sided and tendentious article on Mustine]. PMID- 9349008 TI - [Agreement 1997. Little money in new wage system]. PMID- 9349009 TI - [Health scientific language policy. Formally rule-bound]. PMID- 9349010 TI - [Manuscript guidelines for Sygeplejersken]. PMID- 9349011 TI - [Capital Hospital Community. Limits of pain. Interview by Kirsten Bjornsson]. PMID- 9349012 TI - [HS (Capital Hospital Community)--politicians on some breathing space. Interview by Kirsten Bjornsson]. PMID- 9349013 TI - [HS (Capital Hospital Community)--Copenhagers have more sickness]. PMID- 9349015 TI - [Executive Board--concrete action for a better environment]. PMID- 9349014 TI - [Home from service with a heavy heart. Red Cross Florence Nightingale Medal]. PMID- 9349016 TI - [Executive Board--common goal but also locally freedom to act]. PMID- 9349017 TI - [Executive Board--support prioritizing of social psychiatry]. PMID- 9349018 TI - [Iceland's nurses are academicians]. PMID- 9349019 TI - [Complete victory for HIV-positive nurse]. PMID- 9349020 TI - [Nurses behind unique training--80 percent got leg ulcers within 4 years. Interview by Kjell Arne Bakke]. PMID- 9349021 TI - [Least clinical practice, the most pay. Interview by Marit Fonn]. PMID- 9349022 TI - [Pediatric nurses' viewpoint. Interview by Marit Fonn]. PMID- 9349024 TI - [Women's health--premenstrual syndrome--not a new problem. Interview by Georg Mathiesen]. PMID- 9349023 TI - [Rehabilitation--ambulatory team. Interview by Erik Dale]. PMID- 9349026 TI - [A leader, nurse, patient--now, a director. Interview by Terje Carlsen]. PMID- 9349025 TI - [Close up: Widar Wenaes, nurse supported by justice--following judgment. Interview by Marit Fonn]. PMID- 9349028 TI - [University nursing home--a new concept]. PMID- 9349027 TI - [My workplace: Casefinding Service, Addictive Drugs Secretariat--help for the less fortunate, Oslo. Interview by Marit Fonn]. PMID- 9349029 TI - [From days gone by--operating room nursing]. PMID- 9349030 TI - [Life is not over with a stroke. Interview by Terje Carlsen]. PMID- 9349031 TI - [Cancer--result of unhealthy life style]. PMID- 9349032 TI - [Alternative treatment--we want our own professional group within the NSF. Interview by Kjell Arne Bakke]. PMID- 9349034 TI - [Nursing under a different sky--Cochabamba]. PMID- 9349033 TI - [India--burning of wives--a barbaric custom]. PMID- 9349036 TI - [ICN--world congress with a Scandinavian flavor]. PMID- 9349035 TI - [Professional ethics council--initiative concerning care of the dead]. PMID- 9349037 TI - [Pediatric nursing--care of infants with abstinence syndrome]. PMID- 9349038 TI - [Hospital hygiene--hospital infections should be reduced]. PMID- 9349039 TI - [Sick leave--unequal use of self reporting]. PMID- 9349040 TI - [Treatment of diabetes--fewer restrictions but more individual responsibility]. PMID- 9349041 TI - Urology nurses of Canada: a work in evolution. PMID- 9349042 TI - Estrogen in urinary incontinence treatment: an anatomic and physiologic approach. AB - Most women and health care providers are knowledgeable about the benefits that estrogen replacement therapy has on the prevention of cardiovascular disease and osteoporosis. What is commonly unknown and under research is the role estrogen plays in maintaining continence. The lower urinary tract shares a common embryologic origin with the female genital organs and is hormonally sensitive. Menopause, either surgical or natural, results in decreased or diminished circulating estrogens that can affect the genitourinary system, causing atrophic symptoms. A comprehensive urinary incontinence workup should include assessment of the vaginal mucosa and treatment of hormone deficiency symptoms such as atrophic vaginitis and urethritis. Risk assessment should be done before hormone replacement therapy is considered. PMID- 9349043 TI - The role of vaginal estrogen in the treatment of urogenital dysfunction in postmenopausal women. AB - Decreased estrogen levels result in significantly lower urogenital tract changes and adversely influences quality of life. Consequences include atrophic vaginitis, atrophic urethritis, urinary incontinence, and pelvic organ prolapse. Evaluation of lower genital tract estrogen status is an integral part of evaluating the postmenopausal woman with urogenital symptoms. PMID- 9349044 TI - The psychosocial impact of urinary incontinence on women aged 25 to 45 years. AB - The purpose of this research was to describe the relationship between symptoms of urinary incontinence and their impact on daily activities, and the degree of incontinence-related distress perceived by 25- to 45-year-old women. A second purpose was to identify differences, if any, in impact on daily life and degree of incontinence-related distress perceived among women with stress, urge, and mixed incontinence. Guided by Lazarus and Folkman's (1984) Stress, Appraisal and Coping Theory, a descriptive correlational prospective study (N = 35) was conducted using the Urogenital Distress Inventory and the Incontinence Impact Questionnaire. A significant moderate (r = 0.5701, p = 0.000) correlation was found between urinary incontinence symptoms and their impact on travel, social, physical, and emotional activities. No significant differences were found among women with stress, urge, and mixed urinary incontinence and the impact of incontinence symptoms on their daily activities or with their perceived degree of incontinence-related distress. PMID- 9349045 TI - Re: Local anesthesia for prostate biopsies. PMID- 9349046 TI - Quality of life tools for assessment of urinary incontinence. PMID- 9349048 TI - A data collection system for prostate biopsies. PMID- 9349047 TI - Pharmacologic therapy for acute cystitis in adults: a review of treatment options. AB - This article is a review of the current pharmacologic treatment options for acute cystitis in adults. It is intended to provide the urologic nurse with practical information regarding the administration of the most frequently prescribed antimicrobial drugs. Pharmacologic agents discussed are of the following classes of antimicrobials: beta-lactam antibiotics, fluoroquinolones, sulfonamides, and urinary-specific antiinfectives. The discussion of each drug class includes information about the indications, pharmacodynamics, pharmacokinetics, and recommended dosage regimens. Specific factors that will influence the health care provider's choice of the most appropriate antimicrobial for the treatment of acute cystitis are examined. Key teaching points that should be incorporated into patient education by the nurse are reviewed. PMID- 9349049 TI - Herpes zoster as a reversible cause of neurogenic bladder. PMID- 9349050 TI - Getting ready for certification. Strive for excellence. PMID- 9349051 TI - Circumcision: one of the oldest known surgical procedures. PMID- 9349052 TI - Caring as art and science. PMID- 9349053 TI - Relations in families with a mentally retarded child from the perspective of the siblings. AB - Nowadays most handicapped children in Sweden live with their families. In this study some of the consequences of the normalisation of their lives are analysed, focusing on young siblings of mentally retarded children. Sixteen siblings, aged 5-11, were tested with the Family Relations Test and Kvebaek Family Sculpture Technique, as were siblings in a control group in which each child was chosen to match a sibling in the target group with respect to age and family constellation. The results show few differences between the two groups. According to the Family Relations Test, however, the emotional involvement in the eldest non-retarded sibling differs statistically significant from that of the corresponding child in the control group and especially so with respect to negative incoming feelings. Young siblings also show a tendency to unconsciously place the mentally retarded child at larger distances from themselves than they place the other siblings with respect to the Kvebaek Family Sculpture Technique. The results indicate that siblings of mentally retarded children have another frame of reference when judging family relations compared with that of the children in the control group. Thus siblings of mentally retarded children seem to adapt to change in the family because of the mentally retarded child and qualitatively alter their thinking in relation to the family members. PMID- 9349054 TI - Limitations of the Sense of Coherence Scale in a Swedish Pentecostal population. AB - The Sense of Coherence Scale is used frequently world-wide and is thought to be appropriate to any population. The purpose of this study was to highlight some of the potential limitations of the scale. Fifteen members of the Pentecostal Movement described several difficulties in completing the scale, especially items measuring manageability and comprehensibility. The following three statements indicate the nature of their interpretation of the items and the ambiguity of their answers as they argued that: (1) the concept of 'I' was interpreted as 'I and God', (2) it is not necessary to understand everything in life, because it is enough that God understands, and (3) life is meaningful in itself because of the salvation. According to this interpretation of the items, every proposed answer could be suitable and consequently, the scores can readily be misconstrued. Many of the respondents proposed additions or changes in the wording to make the scale more suitable. The strong ego-centred items seem to be inappropriate for the participants in this study and other populations might also be confronted with similar difficulties. PMID- 9349055 TI - Descriptions of suffering in connection with life values. Healthy individuals' reflections in interviews. AB - Nine semistructured interviews with attendant questions were conducted with the purpose of elucidating how healthy individuals describe suffering and life values in their reflections upon active euthanasia. In order to find the intended meaning in utterances, the interviews were interpreted step by step. The point of departure was the following question: What expressions of suffering and what expressions of life values can be found in the text? A connection was found between the interviewees' descriptions of suffering and life values in their reflections upon active euthanasia. The interviewees who considered close relations to be a value of life expressed suffering as dependence, compassion, violation, abandonment and feelings of guilt, while those to whom health was a value of life expressed suffering as torment, dependence, physical pain, feebleness, hopelessness and dying. Those who saw autonomy as a value of life expressed suffering as dependence and violation and those to whom doing good was a value of life expressed suffering as compassion. When organizing health care and deciding about the response to suffering, it seems important to strive for a response built upon the individual patient's description of suffering and life values. PMID- 9349056 TI - Fathers' experience of childbirth and its relation to crying in his infant. AB - As part of a research project exploring ways through which we can understand the crying infant and its family, this study focuses on the experiences of fathers during labour and delivery of their infant. In a previous part of the project it was shown that fathers' negative experiences during the childbirth were correlated with the amount of crying in the infant during the first months after birth. The aim of the present study was to explore and interpret the experiences that fathers reported in an interview when the infant was between six months and one year of age. A hundred and nine fathers were interviewed. The interviews, which took place in the families' homes and with both parents present, were carried out in dialogue form with open-ended questions. The results reveal that complications during the delivery were significantly correlated with the amount of crying in the infant. Feelings of helplessness, of guilt and that staff behaviour had been negative were more common in the group of fathers who experienced the delivery as a negative event. 'Locus of control' seems to be the most relevant concept. PMID- 9349057 TI - Well-being and functional ability in uraemic patients before and after having started dialysis treatment. AB - In this study perceived well-being and functioning in 28 uraemic patients (14 women and 14 men, mean age 54 years) were measured in the predialysis stage during conservative renal therapy and 3-9 months after having started maintenance dialysis treatment. The patients had participated in a patient education programme in the predialysis stage. Disease-specific symptoms, perceived health (Health Index), functional (SIP) and emotional (STAI) status were analysed. The results showed that there were no significant differences in the patients' correction of uraemia, frequency of symptoms or anxiety prior to and after having started dialysis. After having started dialysis treatment, fatigue, lack of energy and functional disability in work increased while disability in recreation and pastime decreased. Standard bicarbonate correlated significantly to the symptoms of leg cramps and itching. Serum albumin correlated significantly to eating dysfunction in the SIP. There was a large variation within the group with regard to their self-rated disturbances. Some patients reported a relatively intact quality of life, some reported a moderate influence, and some a severe decrease in quality of life irrespective of whether they were in the predialysis state or on maintenance haemodialysis or CAPD. In conclusion, dialysis treatment resulted in increased fatigue and lack of energy, while disease-specific symptoms, functional disability and anxiety did not increase during the first months on dialysis. The symptoms of itching and leg cramps correlated significantly with level of metabolic acidosis, and eating disability correlated with serum albumin levels, indicating that biochemical variables should be combined with patient assessment of health and well-being in order to optimize treatment and care. Moreover, the wide range of scores in all the research variables indicates that assessment of quality of life can be helpful in allocating support to those patients in need of it. PMID- 9349058 TI - The appearance and disappearance of cognitive impairment in elderly patients during treatment for hip fracture. AB - We studied the natural course of cognitive state in 256 consecutive hip fracture patients who were admitted from an independent living situation. We employed a treatment programme that focused on preventing postoperative cognitive impairment. Cognitive function was assessed with the SPSMQ screening test. The incidence of postoperative cognitive impairment among those lucid on admission was 13%, which generally was reversed before discharge. Thirty-seven percent were cognitively impaired on admission; of those, 51% reached normal test scores while in hospital. Those who recovered within the first week had as good a prognosis during the first year as those who remained lucid throughout the hospital stay. Cognitive impairment was associated with an increased complication rate, e.g. a three-fold increase of early fracture displacement and a four-fold increase of wound infection. This increased risk was present even in patients with mild/moderate cognitive impairment and could not entirely be explained by age. Our results suggest that it is possible to decrease postoperative cognitive impairment by routine monitoring of cognitive status, a high level of continuity and a reorientation programme. The routine assessment of the cognitive function is recommended in geriatric patients who are admitted for surgery. PMID- 9349059 TI - Music and other strategies to improve the care of agitated patients with dementia. Interviews with experienced staff. AB - Many patients with dementia symptoms display forms of agitation such as the repeating of words, restlessness and aggression. These forms of behaviour may inflict strain on the co-patients and the caregivers. In this study, 17 experienced formal caregivers from nursing homes and collective residential units were interviewed about their experiences of agitated patients with dementia and strategies to improve their care. The questions were open except for specific questions about sound, music, and opinions about pharmacological treatment. A calm atmosphere and a slow pace emerged as important strategies to control agitation. Fixed routines could develop this. The mixing of lucid and agitated dementia patients appeared as a major problem, because some lucid patients became angry when patients with dementia displayed agitation. Irritability in one patient could trigger agitation in other patients but was possible to stop at an early stage. Several responders had successfully used music to calm individual agitated patients. Music seemed to be an underestimated nursing intervention to control agitation in daily life, but uncontrolled sound could cause agitation in the patients and stress in the nursing staff. PMID- 9349060 TI - Quantitative research on therapeutic touch. An integrative review of the literature 1985-1995. AB - Quantitative research on Therapeutic Touch (TT), published in referred nursing journals from 1985 to 1995, is reviewed. Therapeutic Touch is defined by Dolores Krieger, the founder of this nursing intervention. The authors of this Integrative Review examine what is known and not known to date in order to facilitate appropriate application of this modality in practice, and to offer recommendations for future research. Critical characteristics of eleven quantitative studies are identified and analyzed. These characteristics include: author/year/journal/title; study purpose (hypotheses); background/literature review/conceptual citations; sample selection method; study design/random assignment; independent variable/length of treatment/control and confounders; dependent variables/measurements; outcomes; study limitations; and implications for future research. After reviewing the studies, it is concluded that there is evidence to support the practice of Therapeutic Touch for the reduction of pain or anxiety. There is clearly a lack of congruity between the research statement, conceptual framework, operational definition of TT and the findings. This incongruity is discussed and incorporated in the recommendations for future research including outcome, theory-generating and theory-testing research. PMID- 9349061 TI - Anaesthetic pre-registration house officers. PMID- 9349062 TI - A comparison of the infusion pharmacokinetics and pharmacodynamics of cisatracurium, the 1R-cis 1'R-cis isomer of atracurium, with atracurium besylate in healthy patients. AB - We have compared the pharmacokinetics of cisatracurium with atracurium when given by bolus dose followed by continuous infusion. Twenty healthy patients were anaesthetised with thiopentone, midazolam, fentanyl and 70% nitrous oxide in oxygen. Ten patients (Group C) were randomly allocated to receive cisatracurium 0.1 mg.kg-1 and 10 patients (Group A) were given atracurium 0.5 mg.kg-1. Neuromuscular block was monitored using a mechanomyograph. When the first twitch of the train-of-four had recovered to 5% of control, an infusion of cisatracurium 3 micrograms.kg-1.min-1 was started in Group C and an infusion of atracurium 10 micrograms.kg-1.min-1 was started in Group A. The infusion rates were adjusted to maintain the first twitch of the train-of-four at 5% of control. The times to 90% and maximum depression of the first twitch of the train-of-four were significantly longer after cisatracurium than atracurium (2.2 and 3.4 min compared with 1.3 and 1.8 min, respectively; p < 0.01 in each instance). No significant differences were found in recovery parameters between the two groups. Blood samples were taken at regular intervals following the bolus, during the infusion and for 8 h thereafter. The plasma samples were analysed using high performance liquid chromatography for cisatracurium and atracurium (using a method which distinguishes between the three geometric isomer groups), laudanosine and monoquaternary alcohol. The results were analysed using the Non linear Mixed Effects Model program. A two-compartment model was fitted to the data. The different isomer groups of atracurium have different pharmacokinetics, the trans-trans group having the highest clearance (1440 ml.min-1) and the cis cis group the lowest (499 ml.min-1). The clearance of cisatracurium (425 ml.min 1) is less than that of cis-cis atracurium and its elimination half-life is longer (34.9 min and 21.9 min, respectively). The plasma concentration of laudanosine after cisatracurium was one-fifth of that after atracurium. PMID- 9349063 TI - A comparison of intercuff and single cuff techniques of intravenous regional anaesthesia using 0.5% prilocaine mixed with technetium 99m-labelled BRIDA. AB - Intravenous regional anaesthesia of the upper limb is a widely used technique first described by Bier in 1908. The exact site of action of injected local anaesthetic has not been determined. We have performed intravenous regional anaesthesia on volunteers using prilocaine mixed with technetium 99m-labelled 2,4,6 trimethyl-3-bromo iminodiacetic acid. Two different techniques of intravenous regional anaesthesia (the 'normal' cuff and the intercuff techniques) were combined with gamma camera tracking of the radiolabel to determine the site of local anaesthetic action. The onset of action was similar for both techniques. The local anaesthetic was mainly retained in the antecubital fossa in both techniques but in the 'normal' technique, the local anaesthetic subsequently showed some retrograde spread. This would suggest that the main site of action of local anaesthetic used for intravenous regional anaesthesia is the larger nerves in the vicinity of the antecubital fossa. PMID- 9349064 TI - Effect of nebulised lignocaine on the quality of induction of anaesthesia in cigarette smokers. AB - We have assessed the effect of nebulised lignocaine, given pre-operatively, upon the quality of induction of anaesthesia in cigarette smokers. Seventy-five patients were studied in a double-blind randomised fashion, receiving a nebuliser of either 4 ml 0.9% NaCl or 4 ml 4% lignocaine. All patients received a standardised anaesthetic consisting of thiopentone followed by progressive increments of enflurane. Thirty-three out of 38 patients (87%) who received nebulised lignocaine had induction without adverse events, compared with 25 out of 37 patients (68%) in the nebulised saline group (Chi-squared test p < 0.05). We conclude that the use of nebulised lignocaine, administered pre-operatively, improves the quality of induction of anaesthesia in cigarette smokers. PMID- 9349065 TI - Addition of adrenaline to pethidine for epidural analgesia after caesarean section. AB - We have investigated the addition of adrenaline to epidural pethidine for postoperative analgesia in 40 patients after Caesarean section. In a randomised, double-blind study, patients received pethidine 25 mg with adrenaline 50 micrograms (adrenaline group, n = 20) or pethidine 25 mg without adrenaline (plain group, n = 18) epidurally at the first request for postoperative analgesia. The median duration of analgesia was longer in the adrenaline group (196 min; IQR 123-286) compared with the plain group (96 min; IQR 43-113; p = 0.002) and plasma concentrations of pethidine in the first 30 min after injection were lower in the adrenaline group (p = 0.003). Visual analogue scale pain scores in the first 30 min after injection and onset of analgesia, defined by the time for pain scores to decrease by 50%, were similar between groups. Addition of adrenaline to epidural pethidine has advantages for analgesia after Caesarean section. PMID- 9349066 TI - The addition of opioids to local anaesthetics in brachial plexus block: the comparative effects of morphine, buprenorphine and sufentanil. AB - We compared the duration of analgesia produced by a mixture of lignocaine and bupivacaine, either alone or combined with morphine (75 micrograms.kg-1), buprenorphine (3 micrograms.kg-1) or sufentanil (0.2 microgram.kg-1) in 80 patients after brachial plexus block for orthopaedic surgery of the upper limb. The characteristics of analgesia were evaluated hourly using a visual analogue scale. The analgesia was considered satisfactory for scores of 30 or less. The median duration (range) of satisfactory analgesia was: 11.5 (8-15) h without an opioid, 21 (9-27) h with morphine, 20 (14-34) h with buprenorphine and 24.5 (11 38) h with sufentanil. We conclude that the addition of an opioid to a local anaesthetic mixture lengthens the duration of analgesia. PMID- 9349068 TI - A new hinged tip laryngoscope. AB - The McCoy laryngoscope undoubtedly improves the view of the vocal cords during difficult tracheal intubation. An inherent design problem with the McCoy blade is the need to relax the grip on the laryngoscope handle at the point when stability is most necessary. A new hinged tip blade is described which is operated by a button mechanism on a secondary handle that runs adjacent to the standard handle. This modification offers the possibility of better stability and ease of manipulation whilst maintaining all the benefits of the McCoy blade. PMID- 9349067 TI - The anti-emetic efficacy of a combination of ondansetron and droperidol. AB - The anti-emetic efficacy of a combination of ondansetron 8 mg with either droperidol 0.75 mg or 1.25 mg given prophylactically was studied in a randomised blinded trial of 94 female inpatients with a previous history of postoperative nausea and vomiting and scheduled to have laparoscopic surgery. A standardised general anaesthetic technique was used for all patients. The mean estimated risk of postoperative sickness according to predictive patient characteristics was 65% for both treatment groups. During the 24 h study period, the proportion of patients with nausea was similar (35%) in both groups, and vomiting occurred in 16% and 14% of the patients in the droperidol 0.75 mg and 1.25 mg groups, respectively. No serious adverse events were observed. Ondansetron in combination with droperidol 0.75 mg resulted in less drowsiness than in combination with 1.25 mg (p = 0.03). In conclusion, a prophylactic combination treatment of ondansetron 8 mg with a small dose of droperidol was clinically effective and well tolerated for the prevention of postoperative nausea and vomiting after laparoscopic surgery in patients with a high probability of nausea and vomiting. PMID- 9349069 TI - Additional inspiratory resistance imposed by the laryngeal mask airway: in vitro versus in vivo comparison. AB - The present study was designed: (a) to compare the additional inspiratory laryngeal mask airway (LMA) resistance measured in vitro during simulated ventilation and in vivo in five anaesthetised and mechanically ventilated patients; and (b) to evaluate the resistive pressure drop along the length of the LMA. After the differential pressure across the mask was measured, the pressure flow relationship was characterised by Rohrer's equation and in vitro and in vivo resistance was calculated. Thereafter, the distal pressure measuring point was moved along the length of the LMA and differential pressure was measured at each point under in vitro and in vivo conditions. Values for resistance were approximately twice as great in vivo as those obtained in vitro, with most of the resistive pressure drop occurring across the vertical bars, especially when measured in vivo. We conclude that in vivo positioning of the LMA significantly increases resistance because of the configurational changes occurring when the LMA is in situ. PMID- 9349070 TI - Peri-operative stroke in general surgical patients. AB - The incidence and aetiology of peri-operative stroke in patients undergoing general surgical procedures are reviewed. Some general recommendations on the peri-operative anaesthetic management of these patients are summarised. PMID- 9349071 TI - Cardiac rupture during electroconvulsive therapy. AB - A 57-year-old man with recurrent depression, resistant to drug therapy, was scheduled for a course of eight electroconvulsive therapy treatments. The patient had undergone seven treatments without incident over the previous 3 weeks. Immediately following the final treatment, the patient suffered cardiovascular collapse, culminating in cardiac arrest with electromechanical dissociation. Despite resuscitative measures, the patient died. Post-mortem examination found the cause of death to be cardiac tamponade, secondary to cardiac rupture. PMID- 9349072 TI - Epidural dextran 40 patch for postdural puncture headache. AB - We report a case where two consecutive blood patches failed to relieve postdural puncture headache and hearing loss following inadvertent dural puncture. Initial conservative therapy with analgesics, fluids and later two blood patches had no effect and the patient's symptoms only resolved after an epidural dextran 40 patch performed 8 weeks after the dural tap. Two months later, the patient is still asymptomatic. PMID- 9349073 TI - Subarachnoid block for a case of multiple system atrophy. AB - We report a case of multiple system atrophy in a 60-year-old man who presented twice for optical urethrotomy. Multiple system atrophy is associated with autonomic neuropathy, cerebellar ataxia, disordered control of respiration and paralysis of the abductors of the vocal cords and thus poses several problems for the anaesthetist. Subarachnoid block, which has not been described before in this group of patients, was administered twice and was well tolerated on both occasions. PMID- 9349074 TI - Fatal hepatic necrosis after isoflurane anaesthesia. AB - Several halogenated anaesthetic agents have been associated with hepatotoxicity. We report a case of fulminant, fatal hepatic necrosis after uneventful isoflurane anaesthesia in a patient without previous liver disease, who may have been sensitised by previous exposure to enflurane. Although no anti-trifluoroacetyl antibodies could be detected in the patient's serum, isoflurane hepatotoxicity seems very likely to be the reason for fulminant hepatic failure in this patient. PMID- 9349075 TI - The effect of cricoid pressure and neck support on the view at laryngoscopy. AB - Fifty female patients were studied to compare the view of the larynx at direct laryngoscopy under general anaesthesia with and without cricoid pressure applied. We also compared the view using the standard technique of cricoid pressure with that using cricoid pressure in an upward and backward direction and further investigated whether these views were improved with a firm foam rubber neck support. The order in which the types of cricoid pressure were applied was randomised and also blinded with a drape over the neck. Cricoid pressure was simulated on weighing scales after each case and a mean force of 3.2 kg was applied. The majority of views at laryngoscopy (95%) were grade 1, with too few grade 2 and 3 views for statistical comparison. Both types of cricoid pressure applied without neck support were more likely to give a better view than no cricoid pressure (p < 0.01) and cricoid pressure in an upward and backward direction was more likely to give a better view at laryngoscopy than the standard technique (p < 0.01). Neck support during the standard technique of cricoid pressure did not improve the view of the larynx at laryngoscopy. Cricoid pressure is likely to improve the view at laryngoscopy which may be further improved by applying it in an upward and backward direction. PMID- 9349076 TI - A survey of how British obstetric anaesthetists test regional anaesthesia before caesarean section. AB - We surveyed members of the Obstetric Anaesthetists Association asking how they tested regional blocks prior to Caesarean section. A large proportion of these anaesthetists appear to test their blocks inadequately. Temperature sensation was the most common sensory modality tested (64%) and 79% of those who tested for temperature sensation used an ethyl chloride spray. PMID- 9349077 TI - Taking a 'drugs' history. AB - A simple, anonymous questionnaire was used to examine the depth of drug history taken from patients by a group of junior doctors. The results showed that most who replied restricted their history to prescription medicines and smoking. We feel that this is no longer sufficient and that a drug history should be expanded to include substances such as recreational drugs and over-the-counter preparations. If an allergy is reported, detailed questioning about its nature is also important since the term is often used by patients when a true allergy does in fact not exist. We present a mnemonic which can be used as an aide memoire to the various areas of importance. PMID- 9349078 TI - Experience with the McCoy laryngoscope in difficult laryngoscopy. AB - We report our experience with the McCoy levering laryngoscope in 48 patients who were a Cormack and Lehane grade 3 or grade 4 view at direct laryngoscopy. The view with the blade in neutral position was grade 3 in 39 patients and grade 4 in nine patients. Elevation of the levered tip of the blade in the grade 3 group improved the view to grade 2 in 17 patients (44%), in 17 patients (44%) the view remained unchanged and in five patients (12%) the view deteriorated to grade 4. In the patients initially grade 4, the view improved to grade 3 in one patient and remained unchanged in eight patients. The McCoy laryngoscope is a useful tool to aid intubation in about half of patients who are a grade 3 view at laryngoscopy. Our experience indicates it is unlikely to improve a grade 4 view. PMID- 9349080 TI - Thoracic epidural analgesia started after cardiopulmonary bypass. PMID- 9349079 TI - Prevention of hypotension during spinal anaesthesia for caesarean section: ephedrine infusion versus fluid preload. AB - We compared the efficacy of prophylactic ephedrine infusion over fluid preloading in prevention of maternal hypotension during spinal anaesthesia for Caesarean section. Forty-six women undergoing elective Caesarean section at term were allocated randomly to receive either intravenous fluid preloading with Hartmann's solution 20 ml.kg-1 (fluid group) or prophylactic intravenous ephedrine 0.25 mg.kg-1 (ephedrine group). Moderate hypotension was defined as > or = 20% reduction in systolic blood pressure and severe hypotension as > or = 30% reduction in systolic blood pressure. Maternal uterine circulation was measured using Doppler ultrasound in 11 parturients before and after spinal anaesthesia. There was a lower incidence of severe hypotension in the ephedrine group compared with the fluid group (35% vs. 65%, p = 0.04), although the incidence of moderate hypotension was similar. Mean umbilical venous pH was higher in the ephedrine group than in the fluid group (7.33 vs. 7.29, p = 0.02) and the number of patients shivering was lower in the ephedrine group (2 vs. 9, p = 0.02). No difference was found between pre- and postspinal uterine artery pulsatility indices in either group. We conclude that prophylactic ephedrine infusion alone is at least as good as fluid preload alone in combating the hypotension associated with spinal anaesthesia for Caesarean section. PMID- 9349081 TI - Audit using ICU physiology scoring and outcome prediction. PMID- 9349082 TI - Guidelines and cardiac anaesthetists. PMID- 9349083 TI - Occlusion pressure alarm setting on PCA pumps. PMID- 9349084 TI - Is Hartmann's the solution? PMID- 9349085 TI - An improved Hartmann's solution for anaesthetists in the 1990s? PMID- 9349086 TI - The consultant anaesthetist: UK is OK. PMID- 9349087 TI - Ametop: my view is coloured. PMID- 9349088 TI - Effects of propofol and thiopentone on the immune response. PMID- 9349089 TI - Rocuronium-mivacurium combination is synergistic. PMID- 9349090 TI - Cricoid pressure in chaos. PMID- 9349091 TI - Patient-controlled analgesia. PMID- 9349092 TI - Pseudoaneurysm of the dorsalis pedis artery. PMID- 9349093 TI - Cormack and Lehane revisited. PMID- 9349094 TI - Unilateral pulmonary oedema following postextubation laryngospasm. PMID- 9349095 TI - Identification of the caudal epidural space. PMID- 9349097 TI - Combined spinal-epidural anaesthesia. PMID- 9349096 TI - Gum elastic bougie for difficult double-lumen intubation. PMID- 9349098 TI - Remedy for inspiratory stridor following laryngeal biopsy. PMID- 9349124 TI - Resuscitation. PMID- 9349125 TI - Basic life support. PMID- 9349126 TI - Management of the airway and ventilation during resuscitation. PMID- 9349127 TI - Advanced life support guidelines. European Resuscitation Council. PMID- 9349128 TI - Paediatric and neonatal life support. PMID- 9349129 TI - Drugs in modern resuscitation. PMID- 9349130 TI - Peri-arrest arrhythmias. PMID- 9349131 TI - Fibrillation and defibrillation of the heart. PMID- 9349132 TI - Immersion, near-drowning and drowning. PMID- 9349133 TI - Aspects of resuscitation in trauma. PMID- 9349134 TI - Uniform reporting in resuscitation. AB - The concept of uniform reporting of data in resuscitation has demonstrated its potential value in pre-hospital and in-hospital cardiac arrest in adults, infants and children, in laboratory research, in disaster research and, hopefully, also in trauma care and research. PMID- 9349135 TI - Ethical aspects of resuscitation. PMID- 9349136 TI - Oxygenation criteria--a plea for more information. PMID- 9349137 TI - Lateral positioning for regional analgesia during labour. PMID- 9349138 TI - Interactions between mivacurium and prilocaine. PMID- 9349139 TI - Effects of volume on spinal anaesthesia with 0.25% hyperbaric bupivacaine. PMID- 9349140 TI - Converting pH and H+. PMID- 9349141 TI - "Renal" dopamine. PMID- 9349142 TI - Myopes, squints, and scans. PMID- 9349143 TI - Evaluation of corneal transplantation. PMID- 9349144 TI - Gene therapy for the eye. PMID- 9349145 TI - Corneal ulceration in the developing world--a silent epidemic. PMID- 9349146 TI - New approach in strabismus surgery in high myopia. AB - AIMS: To develop appropriate methods of eye muscle surgery in highly myopic patients with esotropia and hypotropia, with respect to the pathological findings in high resolution magnetic resonance imaging (MRI). METHODS: 35 patients with unilateral or bilateral high myopia and strabismus--that is, axial length of the globe averaged 29.4 mm. Multiple coronal, transverse, and parasagittal MRI image planes were obtained using a Siemens Magnetom 1.5 tesla MRI scanner. In 15 patients with a pathological plane of recti extraocular muscles found by MRI and confirmed intraoperatively, a new technique of eye muscle surgery was performed to re-establish the physiological muscle plane. This was checked postoperatively in addition to the measurement of alignment and motility by MRI. RESULTS: The new MRI finding of a dislocation of the lateral rectus (LR) into the temporocaudal quadrant by 3.4 mm requires new surgical techniques. Only fixing the LR in the physiological meridian at the equator with a silicone loop ('guide pulley') or a non-absorbable suture is a causal therapy. This yields alignment and improves abduction and elevation. CONCLUSIONS: If the described misalignment of the LR is detected by MRI, a common high dosage recess-resect procedure for esotropia may even aggravate the deviation. The most important aim of eye muscle surgery is to normalise the pathological path of the LR. The restoration of the physiological function of the dislocated LR is remarkable. PMID- 9349147 TI - Conclusions of the corneal transplant follow up study. Collaborating Surgeons. AB - AIM: On the basis of finalised data from the Corneal Transplant Follow up Study to identify and quantify factors influencing corneal graft outcome in terms of graft survival, rejection, visual acuity, and astigmatism. METHODS: Multifactorial analysis of 2777 grafts registered by the UK Transplant Support Service from July 1987 to June 1991. RESULTS: Several recipient factors influencing graft survival, rejection, and visual acuity were identified, but no donor factors. Of the operative factors amenable to change, mixed suturing was associated with reduced graft survival, and larger grafts with increased risk of rejection but better visual acuity when surviving. There was increased risk of rejection with poor matching at HLA class I antigens, but mismatched HLA-DR grafts suffered less rejection than those with zero HLA-DR mismatches. Recipient age below 10 years was associated with increased risk of both rejection and graft failure. However, whereas increasing age above 10 years was not associated with differential graft survival, it was significantly associated with decreasing risk of rejection. CONCLUSIONS: While confirming possible benefits of HLA-A and B matching, the expense and delay involved in awaiting matched HLA-DR tissue is unlikely to be justified. Other donor factors are unrelated to graft outcome following screening of tissue by eye banks. The highest rates of graft failure and rejection happen in the early postoperative period, and factors influencing visual outcome are also apparent at this stage. PMID- 9349148 TI - Influence of patient age on refraction and corneal haze after photorefractive keratectomy. AB - AIMS/BACKGROUND: Since wound healing processes are known to be more rapid in those who are young, it was decided to examine the effect of patient age on refractive outcome of photorefractive keratectomy (PRK). METHODS: The records of 599 eyes that had undergone PRK were studied retrospectively. The eyes were categorised by baseline myopia and patient age. Spherical equivalent and corneal haze were compared in the baseline refraction and age groups at 3, 6, and 12 months after PRK. RESULTS: There were no differences in postoperative refraction and corneal haze in the different age groups. CONCLUSION: Patient age had no statistically significant effect on refraction and corneal haze 1 year after PRK. PMID- 9349149 TI - Anterior segment dysgenesis in mosaic Turner syndrome. AB - AIMS/BACKGROUND: Females with Turner syndrome commonly exhibit ophthalmological abnormalities, although there is little information in the literature documenting findings specific to Turner syndrome mosaics. Ophthalmic findings are described in four patients with mosaic Turner syndrome. All had anterior chamber abnormalities and all four had karyotypic abnormalities with a 45, X cell line. The possible relation between the karyotypic and the phenotypic findings in these patients is discussed. METHODS: Four girls with mosaic Turner syndrome underwent a full ophthalmological assessment, including examination under anaesthesia where indicated. RESULTS: Three of the four patients presented with congenital glaucoma. Two had the karyotype 45, X/46, X, idic(Y) and one a 45, X/47, XXX karyotype. The remaining child had a Rieger malformation of the iris and the karyotype 45, X/46, X, r(X). CONCLUSIONS: These findings suggest that Turner syndrome mosaicism (where there are two abnormal cell lines) is associated with anterior segment dysgenesis. The findings in these four patients are compared with those seen in other mosaic phenotypes and it is postulated that the presence of two or more genetically different cell lines may have an adverse effect on anterior segment development. PMID- 9349150 TI - Successful visual rehabilitation after neonatal penetrating keratoplasty. AB - BACKGROUND: Penetrating keratoplasty in infancy and childhood has traditionally met with limited visual success due to a combination of unique physiology and technical problems in this patient population. With the advances in microsurgical instrumentation, corneal preservation, and visual developmental physiology ophthalmologists are finding increasing indications for penetrating keratoplasty in the childhood population. The long term results of neonatal penetrating keratoplasty in two patients with unilateral congenital corneal opacification are reported. METHODS: Penetrating keratoplasty was performed on one eye in each of two infants within the first 3 weeks of life. Amblyopia treatment and optical therapy have been continued since surgery. RESULTS: After 6 years both grafts have remained clear. One patient developed the infantile esotropia syndrome. Visual development using Snellen optotypes is age normal for both transplanted eyes. CONCLUSIONS: Penetrating keratoplasty when combined with optical correction and amblyopia therapy may restore and preserve vision in selected patients with congenital corneal opacification if performed in the neonatal period. PMID- 9349152 TI - Imaging of optic nerve head drusen with the scanning laser ophthalmoscope. AB - BACKGROUND: Optic nerve head drusen may present diagnostic difficulties in cases of disc swelling. Imaging of the nerve in a search for drusen is often inconclusive, especially in children, where drusen may be buried below the surface of the nerve head. METHODS: A small study was carried out using a scanning laser ophthalmoscope (SLO) with an infrared confocal facility to scan deep within optic discs in an attempt to image drusen. RESULTS: The SLO was able to demonstrate superficial and buried drusen (using the infrared confocal facility). The superiority of the SLO over ultrasound in the presence of lens opacity was revealed, as the SLO simultaneously demonstrated both drusen and the associated anomalous disc features which are not detected by ultrasound. CONCLUSION: The SLO can help in the diagnosis of optic disc drusen especially in difficult cases where lens opacity or buried drusen hinders their definitive diagnosis. PMID- 9349151 TI - Primary association of HLA-B51 with Behcet's disease in Ireland. AB - AIMS/BACKGROUND: While a primary association of HLA-B51 with Behcet's disease (BD) in Japanese and Mediterranean patients supports an immunogenetic predisposition, this link is unclear in north western Europe. This study assessed HLA associations with BD, and HLA-B51 with certain clinical characteristics, in the Republic of Ireland, which has an ethnically homogeneous population. METHODS: HLA-A, HLA-B, and HLA-DR typing was performed in 24 BD patients, conforming to International Study Group criteria, and in blood donors, as controls. Patient records were retrospectively reviewed and patients reassessed clinically. RESULTS: A highly significant HLA-B51 association (corrected exact p value = 0.002, relative risk = 6.3) with BD was determined, despite a low B51 prevalence (25%) in patients. No other HLA type was associated. There was a significant B51 link with male sex in BD patients but no association with age at first manifestation/diagnosis, eye involvement, cyclosporin A therapy, or poor visual acuity was determined. CONCLUSIONS: This study supports a HLA-B51 immunogenetic predisposition, similar to Japanese patients, in Irish BD in an ethnically homogeneous population in north western Europe. However, owing to a low prevalence of B51 positivity in BD patients in Ireland, a multifactorial pathogenesis is suggested. PMID- 9349153 TI - Late reopening of successfully treated macular holes. AB - BACKGROUND: Most idiopathic macular holes can be closed by a surgical procedure combining vitrectomy, posterior hyaloid ablation, and fluid-gas exchange followed by postoperative positioning. Reopening of closed macular holes has been reported, but its frequency is not known. Here the incidence of reopening after successful macular hole surgery is reported. METHODS: 77 consecutive cases of idiopathic macular holes operated with autologous platelet injection between July 1993 and October 1995 were reviewed. The procedure consisted of three port vitrectomy, posterior hyaloid removal, nonexpansile fluid-gas exchange, and autologous platelet injection followed by face down positioning. The incidence of reopening was analysed in the cohort of the 72 anatomical successes. RESULTS: Mean follow up was 12.3 months. The macular hole reopened in five eyes of five patients (five out of 72 patients, 6.9%), in four cases after cataract extraction. In four cases too, an epiretinal membrane was noted, either clinically or during reoperation, and fluorescein leakage in the macular area was present in two cases. Three of the five cases of reopening were reoperated and all three were anatomical successes. CONCLUSION: Late macular hole reopening occurred in five out of 72 patient, and in four cases after cataract surgery. The presence of an epiretinal membrane around the hole in four of them suggested that tractional forces were responsible for the reopening. Reoperation, performed in three cases, again closed the macular holes. PMID- 9349154 TI - Peripapillary circle of Zinn-Haller revealed by fundus fluorescein angiography. AB - AIMS: To observe the vascular pattern of the peripapillary circle of Zinn-Haller in humans by fundus fluorescein angiography. METHODS: 307 cases (from 212 patients) of fundus fluorescein angiograms performed in patients with myopic degeneration were evaluated to find the circle of Zinn-Haller and to observe its fundus fluorescein angiographic features. RESULTS: 15 cases (from 13 patients) with the circle of Zinn-Haller were found. It appeared as concentric or zigzag shaped vascular fillings within the temporal crescent region. All cases were observed in pathological myopia with peripapillary atrophy and a tilted disc. Each arterial circle showed variations in location and shape. CONCLUSIONS: The temporal part of the circle of Zinn-Haller can be revealed by fundus fluorescein angiography particularly in pathological eyes with prominent peripapillary atrophy and a tilted disc. The morphological variation of this arterial circle should be considered. PMID- 9349155 TI - Syringomatous carcinoma of the eyelid and orbit: a clinical and histopathological challenge. AB - AIMS: To present three patients with a syringomatous carcinoma (SC). SC is a rare cutaneous neoplasm, most frequently situated on the face and scalp and histologically characterised by an infiltrative pattern of basaloid or squamous cells, a desmoplastic stromal reaction, keratin filled cysts, and granular structures. METHODS: The clinical histories of the patients with a SC were investigated retrospectively. RESULTS: Patient 1 had a benign appearing tumour of the lower eyelid. Five tumour excisions were necessary to remove the SC completely. Patient 2 had a tumour on the lateral part of the lower eyelid and in the medial canthal area. The histopathological findings revealed a squamous cell carcinoma, later revised as a SC. In spite of two excisions and one microscopically controlled excision, a recurrence occurred. An exenteration orbitae was recommended. Patient 3, known to have a history of multiple malignant skin tumours after kidney transplantation and use of cyclosporin, presented with a firm mass in the eyebrow region and in the nasal area of the orbit. The pathological diagnosis of this adnexal tumour was difficult. An exenteration was recommended. CONCLUSIONS: SC is a benign appearing but extremely invasive, locally destructive, slowly growing adnexal tumour, derived from eccrine sweat glands. It is often mistaken, both clinically and microscopically, for other benign and malignant entities. The tumour recurrence is high due to extensive perineural invasion, but regional or distant metastases are rare. The local aggressive nature of the tumour and the high recurrence rate may necessitate mutilating procedures. Optimal treatment consists of a complete microscopically controlled surgical excision with clear surgical margins. PMID- 9349156 TI - Sub-Tenon's anaesthesia: an efficient and safe technique. AB - AIM: To evaluate sub-Tenon's anaesthesia as an alternative to peribulbar anaesthesia. METHODS: 109 consecutive patients listed for various eye operations (including cataract, trabeculectomy, and vitrectomy) under peribulbar anaesthesia were operated on under sub-Tenon's anaesthesia instead. After topical anaesthesia a buttonhole was fashioned through the conjunctiva and Tenon's capsule 10 mm posterior to the limbus. 1.5 ml of lignocaine 2% was then delivered to the posterior sub-Tenon's space using a blunt cannula. The surgical procedure was performed immediately after the completion of the anaesthetic procedure. Chemosis, conjunctival haemorrhage, degree of akinesia, and pain scoring were analysed. RESULTS: There were no anaesthesia related complications. The administration of the block was painless for 99.1% of the patients. In all, 97.3% reported no pain during surgery. There was no akinesia when assessed just after the completion of the block and akinesia was limited when assessed after surgery. Chemosis and conjunctival haemorrhage were frequent but caused no intraoperative problems. CONCLUSION: Sub-Tenon's anaesthesia is an efficient and safe anaesthetic technique. It is a good alternative to peribulbar anaesthesia. PMID- 9349157 TI - Tenascin-C expression in normal, inflamed, and scarred human corneas. AB - AIMS/BACKGROUND: In adult tissues the expression of tenascin-cytotactin (TN-C), an extracellular matrix glycoprotein, is limited to tumours and regions of continuous renewal. It is also transiently expressed in cutaneous and corneal wound healing. There are limited data regarding its expression in inflammation and scarring of the adult human cornea. In this study, TN-C expression patterns in normal, inflamed, and scarred human corneas have been examined. METHODS: Penetrating keratoplasty specimens were selected from cases of herpes simplex keratitis, herpes zoster ophthalmicus, rheumatoid arthritis ulceration, bacterial keratitis, rosacea keratitis, interstitial keratitis, and previous surgery so as to encompass varying degrees of active and chronic inflammation and scarring. TN C in these and in normal corneas was immunodetected using TN2, a monoclonal antibody to human TN-C. RESULTS: There was no TN2 immunopositivity in normal corneas except at the corneoscleral interface. In pathological corneas, TN2 immunopositivity was localised in and around regions of active inflammation, fibrosis, and neovascularisation. TN2 positivity was less in acute inflammation than in active chronic inflammation. Mature, avascular scar tissue and epithelial downgrowth were TN2 negative. CONCLUSION: These results indicate that in the adult human cornea, TN-C expression is induced in regions of inflammation, fibrosis, and neovascularisation, but that expression is absent in mature, avascular scar tissue. This suggests a role for this glycoprotein in inflammation, healing, and extracellular matrix reorganisation of the cornea. PMID- 9349158 TI - Impression cytology in Down's syndrome. AB - AIM: To evaluate both the number and the average distribution of goblet cells, which are responsible for the production of the mucin layer of the tear film, in the bulbar conjunctiva of patients with Down's syndrome. Previous research had used the ferning test to indicate an alteration in Down's syndrome, but had not determined which film layer was involved. METHODS: The presence of goblet cells in the bulbar conjunctiva of 30 subjects (15 with Down's syndrome, and 15 normal control subjects) was evaluated using impression cytology. RESULTS: A marked reduction of goblet cells was found in the Down's syndrome group (81.4 per mm2) when compared with the control group, where (209.8 per mm2) was found. CONCLUSION: The deficit observed appears to be the cause of the tear film alterations observed in Down's syndrome. In turn, this may often lead to the formation of dry spots, and to consequent frequent infections of the anterior segment of the eye. While it is further hypothesised that the alteration of the conjunctival epithelium in Down's syndrome may be due to an altered metabolism of some element or elements, such as vitamin A, further research will be necessary to corroborate this. PMID- 9349159 TI - Effect of beam variables on corneal sensitivity after excimer laser photorefractive keratectomy. AB - AIM: To investigate changes in corneal touch sensitivity following excimer laser photorefractive keratectomy (PRK) using different beam configurations. METHODS: 20 subjects were given a unilateral -3.00 D correction with either a 5 mm (26 micrograms, n = 10) or 6 mm (42 micrograms, n = 10) beam diameter. Thirty subjects underwent a unilateral -6.00 D correction with 5 mm (62 micrograms, n = 10), 6 mm (78 micrograms, n = 10), or multizone (62 micrograms, n = 10) treatments. The multizone treatment was 6 mm in diameter with the depth of the 5 mm treatment. Corneal sensitivity was measured using a slit-lamp mounted Cochet Bonnet aesthesiometer before and at 1, 3, 6, and 12 months after PRK. Stimulus locations included points lying within the ablated zone (central) and outside (peripheral). These were compared with the equivalent locations in control (untreated) eyes. RESULTS: There was a significant reduction in corneal sensitivity within the central (ablated) zone in all treatment groups after PRK. In most groups a return to full sensitivity was achieved by 6 months with the exception of the multizone treatment group which showed significant corneal hypoaesthesia at 12 months. Peripheral corneal sensitivity was also reduced in this group up to 3 months after the procedure. A comparison between the -3.00 D and -6.00 D treatment groups showed no significant difference. However, combining data from all treatment groups, a significant correlation was found between the interocular difference in central corneal sensitivity and postoperative haze at 3 and 6 months. CONCLUSIONS: For corrections up to -6.00 D ablation depth and treatment zone diameter do not appear to be clinically important determinants of corneal hypoaesthesia. In contrast, postoperative corneal haze appears to correlate with sensitivity loss. PMID- 9349160 TI - Retinoinvasive malignant melanoma of the uvea. AB - AIMS: To define a retinoinvasive phenotype of uveal melanoma based on an informative case and survey of literature. METHODS: A 65-year-old woman developed a circumscribed mixed cell type melanoma of the ciliary body that was locally excised. After 6 years, secondary glaucoma evolved. Three years later a ring melanoma was diagnosed and the eye was enucleated. The histopathological material was analysed by immunohistochemistry. RESULTS: A spindle cell type ring melanoma infiltrated the iris and ciliary body diffusely, and extended through the aqueous outflow channels and iridocyclectomy flap extrasclerally. The choroid was uninvolved. Instead, tumour cells spread to the vitreous and along the ciliary epithelium, adhered to the hyaloid face and retinal surface, and extensively invaded the neuroretina, the retrobulbar optic nerve, and perineural space. They were labelled for S-100 protein, vimentin, and in the neuroretina for cytokeratins 8 and 18. No evidence of systemic disease is evident 5 years after enucleation. Three identical tumours of the iris and ciliary body that extensively infiltrated the neuroretina and retrobulbar optic nerve were identified from previous literature. CONCLUSION: Retinoinvasive melanoma is a rare but distinct phenotype of uveal melanoma, different from circumscribed and most diffuse melanomas that may erode the overlying retina and infiltrate the optic nerve that do not invade non-adjacent retina. Retinoinvasive tumours tend to evolve from a ring melanoma and they grow slowly, which may favour emergence of tumour clones able to migrate, adhere to, and invade into the neuroretina, analogous to the metastatic cascade. Frequent secondary angle closure glaucoma may promote invasion into the optic nerve. PMID- 9349162 TI - Visual recovery from no light perception in total retinal detachment with massive subretinal haemorrhage. PMID- 9349163 TI - Bilateral angle closure glaucoma: an unusual presentation of Vogt-Koyanagi-Harada syndrome. PMID- 9349164 TI - Ultrasound biomicroscopy of angle closure glaucoma with pseudoexfoliation syndrome. PMID- 9349165 TI - The cotton bud as a missile causing penetrating eye injury. PMID- 9349166 TI - Ankyloblepharon filiforme adnatum. PMID- 9349161 TI - Central retinal vein occlusion: what's the story? PMID- 9349167 TI - Discoid corneal oedema and high intraocular pressure following PRK. PMID- 9349168 TI - Coeliac disease and Behcet's disease. PMID- 9349225 TI - Diagnostic discord with melanoma. AB - The study of the Pathology Panel of the Dutch Melanoma Working Party highlights the great difficulty in achieving uniform diagnostic assessment of melanoma. Their solution is to set up a national reference panel and to focus continuing medical education on identified areas of particular difficulty. This could be appropriate for other countries, although selection of referees and funding may be problematic. It may also be timely to consider whether melanoma terminology can be rationalized to make it more likely to be reported consistently by pathologists whilst still providing sufficient information for proper patient management. Alternatively the reporting of most melanocytic lesions could be confined to pathologists who specialise in this subject, a practice which has evolved for other areas of pathology. This would facilitate the maintenance of standards and uniformity among that smaller group, but it would not avoid the need for a continuing awareness among all pathologists of the diagnostic pitfalls which abound in the area of melanocytic lesions. PMID- 9349226 TI - In situ hybridization and its diagnostic applications in pathology. AB - In situ hybridization (ISH) is a technique by which specific nucleotide sequences are identified in cells or tissue sections. These may be endogenous, bacterial or viral, DNA or RNA. On the basis of research applications, the technique is now being translated into diagnostic practice, mainly in the areas of gene expression, infection and interphase cytogenetics. Diagnostic applications are most often based on short nucleotide sequences (oligomers) labelled with non isotopic reporter molecules, and sites of binding may be localized by histochemical or immunohistochemical methods. The technique can be applied to routinely fixed and processed tissues; with some targets, it is even possible to obtain hybridization in autopsy material. ISH has been used to detect messenger RNA (mRNA) as a marker of gene expression, where levels of protein storage are low; for example, to confirm an endocrine tumour as the source of excess hormone production. Its application in infectious diseases has to date been mainly in viral infections, such as the typing of human papillomavirus (HPV) or the detection of Epstein-Barr virus by the presence of small nuclear RNAs (EBERs). The expression of mRNAs for histone proteins has been used to detect cells in S phase, and related methods may be applied to detect apoptotic cells. Using probes to chromosome-specific sequences, it is possible to detect aneuploidy, and to document changes in specific chromosomes, which may have prognostic significance in some tumours, such as B-cell chronic lymphatic leukaemia. Using sequence specific probes, translocations can be identified, such as the t(11;12) of Ewing's sarcoma. This review presents an outline of the technique of in situ hybridization and discusses areas of current and potential diagnostic application. PMID- 9349227 TI - Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV8)--a new human tumour virus. AB - Recently, a novel DNA virus has been molecularly cloned from Kaposi's sarcoma (KS) tissue, a tumour common in acquired immune deficiency syndrome (AIDS). Analysis of the viral genome confirms that it is a relative of human herpesviruses and the virus has been designated HHV-8. Epidemiological evidence suggests a strong aetiological link between the presence of HHV-8 DNA and/or antibodies against the virus, and KS. Additional sequence analysis suggests that the HHV-8 genome contains sequences which encode a D type cyclin and a number of other genes potentially implicated in growth deregulation which may be relevant to its proposed role as a transforming virus. PMID- 9349228 TI - Quality assessment by expert opinion in melanoma pathology: experience of the pathology panel of the Dutch Melanoma Working Party. AB - Some cutaneous melanocytic lesions are notoriously difficult to diagnose by histopathology. For that reason, the Pathology Panel of the Dutch Melanoma Working Party was instituted and is regularly approached to provide an expert opinion on problem cases. In order to identify the most common diagnostic problems, 1069 consecutive referral cases of submitted lesions (1992 to 1994 inclusive) were analysed. About 60 per cent of the requests came from small laboratories, with up to three consultant pathologists. Two-thirds of the lesions reviewed concerned women and nearly 50 per cent of the patients were 30 years of age or younger. In 8 per cent of the cases, the referring pathologists felt unable to make a confident diagnosis; in 14 per cent, melanoma was suspected; and in 12 per cent, a differential diagnosis only had been formulated. The panel felt able to provide an unequivocal diagnosis in 93 per cent of the requests. Of the 158 lesions classified as 'invasive melanoma' by the referring pathologists, 22 were considered to be benign by the panel. Conversely, 108 invasive melanomas (panel diagnosis) had originally been considered as benign lesions, dysplastic naevi or melanoma in situ. These high numbers of discordancies reflect the intrinsic difficulty of the differential diagnoses in this selected material submitted to the panel. Diagnostic difficulties were most often encountered with Spitz naevi and dysplastic naevi. Although the rate of overdiagnosis and underdiagnosis is quite high, in the majority of cases the diagnosis of the referring pathologist matched the diagnosis of the panel. This may reflect a proper awareness of difficult melanocytic lesions in pathology practice. The activities of the Pathology Panel of the Dutch Melanoma Working Party contribute to the improvement of the quality of diagnosis in cutaneous melanocytic lesions, as they increase the number of unequivocal diagnoses and reduce the number of incorrect diagnoses. On the basis of the systematic comparison of the diagnosis by the referring pathologist and the panel, postgraduate teaching and quality control can be more focused on specific diagnostic problems. PMID- 9349229 TI - Ultrastructural characterization of human herpesvirus 8 (Kaposi's sarcoma associated herpesvirus) in Kaposi's sarcoma lesions: electron microscopy permits distinction from cytomegalovirus (CMV). AB - Kaposi's sarcoma (KS) has been shown by molecular techniques to be associated with infection with human herpesvirus 8 (HHV8/KSHV), but specific ultrastructural characterization of the virus has been impaired by the frequent presence in these lesions of other herpesviruses, particularly cytomegalovirus (CMV). Since the ultrastructural appearance of HHV8/KSHV has been studied in the cell line KS-1 uninfected with other viruses including CMV, it was possible to undertake a comparative study of CMV and HHV8/KSHV in KS lesions. HHV8/KSHV was sparsely present and lytic infection was restricted to endothelial cells. The following specific ultrastructural features allowed distinction between HHV8/KSHV and CMV: the viral particles were more delicate and less numerous in cases of HHV8/KSHV infection; the viral tegument was more electron-dense in CMV than in HHV8/KSHV; dense bodies characteristic of CMV were absent in HHV/KSHV; complete CMV viral particles were more variable in size and generally larger (150-200 nm) than HHV8/KSHV (120-150 nm); and finally, the viral envelope was more pleomorphic in CMV than in KSHV/HHV8. Similarities between CMV and HHV8/KSHV included the basic structure of the nucleocapsids and the presence of capsids lacking central DNA cores (so-called non-infectious enveloped particles). These observations show that electron microscopy can be used to identify HHV8/KSHV and confirm the relationship between HHV8/KSHV and KS. PMID- 9349230 TI - The polymerase chain reaction in the diagnosis of early mycosis fungoides. AB - The earliest or patch stage of mycosis fungoides may present diagnostic difficulty both clinically and pathologically. The present study of the polymerase chain reaction (PCR) as a diagnostic tool in early mycosis fungoides was therefore undertaken, using a rapid PCR method for the detection of gamma- and beta-chain T-cell receptor (TCR) gene rearrangements in routine formalin fixed, paraffin-embedded histological sections. Forty-two biopsies were studied from 26 patients with mycosis fungoides. Twenty-three skin biopsies with a clinicopathological diagnosis of early, or patch stage, mycosis fungoides were investigated. Of these, 18 (78 per cent) showed TCR-gamma or both beta- and gamma chain TCR gene rearrangements. TCR gene rearrangements were shown in seven of the 14 plaque stage lesions (50 per cent) and also in the single case of tumour stage disease. Where gene rearrangements were identified, these were identical in all biopsies from an individual patient, irrespective of the site of the lesion, the disease stage, or the time lapse between the biopsies. The PCR is therefore a highly sensitive technique, which can be performed on routine pathological material, in cases where the diagnosis of early mycosis fungoides cannot be made with certainty on conventional histopathological and immunohistochemical grounds. PMID- 9349231 TI - The mummified Hodgkin cell: cell death in Hodgkin's disease. AB - This study attempts to define more clearly the morphology and ultrastructure of mummified Hodgkin cells, to determine their incidence in the different histological subtypes of Hodgkin's disease (HD), and to correlate these data with the expression of p53, bcl-2, mdm2, and p21/WAF1. Forty-five cases of primary HD were examined at light and electron microscopic level. DNA strand breaks were detected by the in situ end-labelling (ISEL) and the TdT-mediated dUTP digoxigenin nick end-labelling (TUNEL) technique. Mummified Hodgkin cells display morphological features that differ from those of classical apoptosis. In contrast to apoptotic cells, mummified Hodgkin and Reed-Sternberg (HRS) cells do not react in the ISEL or TUNEL procedures and maintain the expression of antigens such as CD30 and CD15. The morphology of mummified tissue cells could be simulated by CD95-mediated induction of apoptosis in the Hodgkin cell line HDLM2 if internucleosomal DNA fragmentation was inhibited by zinc ions. The highest incidence of mummified cells was found in the nodular sclerosis and mixed cellularity subtypes, whereas the lowest frequency was observed in nodular paragranuloma. The frequency was independent of p53, bcl-2, p21, and mdm2 expression. p21 and mdm2 immunoreactivity of HRS cells was correlated with p53 status. HRS cells in nodular paragranuloma were virtually negative for p21/WAF1 or bcl-2. Classical apoptotic cells reacting in the TUNEL and ISEL procedures are found in all subtypes of HD and are derived from the non-neoplastic cellular background. In conclusion, mummified Hodgkin cells display features of apoptosis lacking the internucleosomal DNA fragmentation. The pattern of the p53 transactivated genes mdm2 and p21/WAF1 suggests that inactivating mutations of p53 are rare in HD. PMID- 9349232 TI - Modulation of interleukin-6 expression in Hodgkin and Reed-Sternberg cells by Epstein-Barr virus. AB - Variable proportions of Hodgkin's disease (HD) cases are associated with the Epstein-Barr virus (EBV), but the role of EBV in HD is not entirely clear. Hodgkin and Reed-Sternberg (HRS) cells of EBV-associated HD are characterized by expression of the EBV gene product LMP1. In other cellular environments, LMP1 has been shown to induce interleukin (IL)-6. In this study, 105 HD cases were tested for differences in IL-6 expression among LMP1-positive and -negative cases. Isotopic in situ hybridization and correlation with the presence of EBV gene products revealed significantly higher proportions of cases with IL-6-expressing tumour cells in LMP1-positive (31 of 37, 84 per cent) as compared with LMP1 negative HD cases (35 of 68, 51 per cent). Thus, although not exclusive to EBV positive HRS cells, IL-6 expression appears to be upregulated in EBV-associated HD. IL-6 receptor (CD126) expression was tested by in situ hybridization and found in a broad spectrum of cell types, regularly including HRS cells. Superinduction of IL-6 expression may be among the mechanisms by which EBV confers a growth advantage on virus-infected HRS cells and by which the virus may contribute to the morphological and clinical peculiarities of HD. PMID- 9349233 TI - bcl-2 and p53 protein expression in follicular lymphoma. AB - Recent studies have shown bcl-2 to be regulated by p53. Other studies have suggested an inverse relationship between p53 and bcl-2 protein expression in breast and colonic cancers and in a variety of subtypes of non-Hodgkin's lymphoma. This study investigates the relationship between bcl-2 and p53 protein expression and the correlation between these findings and the grade and cell type of follicular lymphomas according to the REAL classification. Paraffin-embedded nodal follicular lymphomas (n = 37) were subjected to bcl-2 and p53 immunohistochemistry on tissue sections using a three-step ABC system. Positive immunostaining for both oncoproteins was scored using a three-tiered scale: +, < 10 per cent cells; ++, 10-50 per cent cells; and ++(+), > 50 per cent cells (< 10 per cent was used as a cut-off to define negative tumours). Ninety-seven per cent (36/37) of follicular lymphomas expressed bcl-2 protein in all three grades, manifesting in the small cell (grade 1) through to the large cell (grade 3). p53 protein expression showed a pattern of increasing immunostaining with progression towards the high-grade follicular lymphoma: grade 1 = 6 per cent (1/16); grade 2 = 48 per cent (10/21); grade 3 = 100 per cent (6/6). Five cases comprised varying combinations of grades. This latter finding suggests a role for p53 mutation in the progression/transformation of follicular lymphoma. The mechanism, however, differs from that suggested in breast and colonic cancers, since an inverse relationship between bcl-2 and p53 was not demonstrated in the present study. PMID- 9349234 TI - Expression of cytotoxic molecules in intestinal T-cell lymphomas. The German Study Group on Intestinal Non-Hodgkin Lymphoma. AB - Intestinal T-cell lymphoma (ITCL) represents a subgroup of peripheral T-cell lymphomas which is thought to arise from alpha beta intraepithelial T lymphocytes. Since these lymphocytes may contain cytotoxic molecules, the question of whether this also holds true for ITCL arises. Twenty ITCL cases were examined for the presence of granzyme B, perforin, and T-cell-restricted intracellular antigen (TIA-1)/granule membrane protein of 17 kD (GMP-17). Two molecules with restricted expression in cytotoxic cells, granzyme B and perforin, were detected by immunocytochemistry and by in situ hybridization with an isotopically labelled RNA probe, respectively. Immunocytochemistry was also performed with the antibody 2G9, which recognizes two molecules, one expressed by cytotoxic cells (TIA-1) and the other found in granulocytes and cytotoxic cells (GMP-17). Granzyme B, TIA-1/GMP-17, and perforin were found in the neoplastic cells of 16/19 cases, 19/20 cases, and 16/17 cases, respectively, of ITCL, but not in the tumour cells of the control group, which consisted of intestinal B cell lymphomas (five cases) and CD8-negative peripheral nodal T-cell lymphomas (six cases). At least one of these molecules was expressed in the tumour cells of all ITCL cases. 2G9 proved to be the most sensitive immunohistological marker, since reactivity with this antibody was not only observed in the highest number of cases, but also found in high numbers of neoplastic cells in positive cases. In conclusion, ITCL appears to show cytotoxic differentiation in all cases. In conjunction with immunophenotypic and genotypic data, our results support a uniform derivation of this tumour from intraepithelial alpha beta cytotoxic T lymphocytes. PMID- 9349235 TI - Prognostic value of tumour-infiltrating lymphocytes (TILs) in colorectal cancer. AB - Clinical follow-up data of 276 colorectal adenocarcinoma patients treated in Kuopio University Hospital between 1976 and 1986 and followed up for a mean of 14 years were analysed. The clinical findings were correlated with tumour infiltrating lymphocytes (TILs) and with histological and quantitative factors including nuclear parameters and volume-corrected mitotic index. In univariate survival analysis, TNM classification, Dukes' stage, histological grade, and TILs were significant predictors of survival. TNM classification, Dukes' stage, and TILs also predicted recurrence-free survival. In multivariate analysis, TILs were an independent prognostic factor of survival in all cases, as well as in patients with T1-4N0-3M0 and T1-4N1-3M1. TILs also independently predicted recurrence-free survival. TILs can provide important prognostic information in colorectal cancer to be used in evaluating for adjuvant therapy in different tumour stages. PMID- 9349236 TI - Patterns of alpha- and beta-catenin and E-cadherin expression in colorectal adenomas and carcinomas. AB - Previous in vitro and in vivo model studies have shown that when E-cadherin expression in carcinoma cells is reduced, invasive behaviour ensues. The situation in human cancer in vivo, however, appears to be more complex, as immunohistochemically determined E-cadherin expression in various carcinomas, including colorectal cancer, does not always correlate with invasive growth. Loss of cell adhesion during invasion in spite of E-cadherin expression might be associated with a defective cadherin-catenin complex. The expression of alpha- and beta-catenin in comparison with E-cadherin was therefore examined in colorectal adenomas and carcinomas and in lymph node and liver metastases. In normal colonic mucosa, alpha- and beta-catenin immunoreactivity occurred along the lateral plasma membranes of the epithelial cells, in a pattern identical to E cadherin staining. A similar pattern was found in colorectal adenomas and in most malignancies. In general, in neoplastic epithelia, the majority of the cancer cells displayed a normal (matching) pattern of E-cadherin and catenin expression. It is concluded that the patterns of expression of E-cadherin and alpha- and beta catenin are very similar in colorectal neoplasms. This observation indicates that invasion in colorectal cancer is not paralleled by consistent loss of expression of the components of the cadherin-catenin complex. PMID- 9349237 TI - Reduced expression of the cadherin-catenin complex in oesophageal adenocarcinoma correlates with poor prognosis. AB - The E-cadherin-catenin complex is important for cell-cell adhesion of epithelial cells. Impairment of one or more components of this complex is associated with poor differentiation and increased invasiveness of carcinomas. Oesophageal adenocarcinomas causes early metastases, progress rapidly, and consequently have a poor prognosis. By means of immunohistochemistry, the expression of E-cadherin and alpha- and beta-catenin was studied in 65 oesophageal adenocarcinomas and 15 lymph node metastases. Expression of these proteins was evaluated with respect to clinico-pathological parameters and patient survival. Expression of the proteins was strongly correlated. In carcinomas, reduced expression of E-cadherin, alpha catenin, and beta-catenin was found in 74, 60, and 72 per cent, respectively. Expression of E-cadherin and alpha-catenin correlated significantly with stage and grade of the carcinomas, whereas expression of beta-catenin correlated only with grade. Reduced expression of all three proteins correlated with shorter patient survival. In contrast to grade, E-cadherin and beta-catenin were significant prognosticators for survival, independent of disease stage. We conclude that in oesophageal adenocarcinomas, decreased expression of E-cadherin, alpha-catenin and beta-catenin are related events. Furthermore, expression of at least E-cadherin and beta-catenin is significantly correlated with poor prognosis. PMID- 9349238 TI - Endocrine cell growths in atrophic body gastritis. Critical evaluation of a histological classification. AB - The aim of the present study was to evaluate the correspondence of the classification of non-antral endocrine cell growths proposed by Solcia and co workers with clinical features and non-endocrine mucosal changes. For this purpose, 94 cases of newly diagnosed atrophic body gastritis were investigated using endoscopic biopsies and compared with 18 control subjects. The patients were subdivided into the following four groups according to the most severe pattern of endocrine cell proliferation found in the body mucosa, as shown by chromogranin A immunostaining: group 1, normal pattern (7 cases, 7.5 per cent); group 2, simple hyperplasia (6 cases, 6.5 per cent); group 3, linear hyperplasia (24 cases, 25.8 per cent); group 4; micronodular hyperplasia (56 cases, 60.2 per cent). Adenomatoid hyperplasia was found in only one case, thus precluding further analysis. Patients in groups 1 and 2 had lower acid secretion, higher gastrin level, and higher mean scores in all histopathological variables of chronic gastritis considered by the Sydney system when compared with controls, but did not differ among them in any parameter investigated. When compared with groups 1 and 2, patients of groups 3 and 4 showed higher values of circulating gastrin, higher scores of glandular atrophy, and lower values of acid secretion and of mononuclear and neutrophil inflammatory cell infiltration. Moreover, group 4 patients differed significantly from those of group 3 in their higher gastrin levels and atrophy scores, and lower scores of neutrophil cell infiltration. On the basis of these results, it is proposed that for practical purposes the normal and the simple hyperplasia patterns may be incorporated into a single group. It is concluded that this classification in its simplified form, based on a qualitative histological approach, shows clinical relevance without the need to perform expensive, time-consuming morphometric evaluations. PMID- 9349239 TI - Imbalance of expression of matrix metalloproteinases (MMPs) and tissue inhibitors of the matrix metalloproteinases (TIMPs) in human pancreatic carcinoma. AB - Degradation of the extracellular matrix (ECM) is an essential step in tumour invasion and metastasis. The matrix metalloproteinases (MMPs) each have different substrate specificities within the ECM and are important in its degradation. MMP activity is dependent on the levels of activated MMP and tissue inhibitors of matrix metalloproteinases (TIMPs). The expression of MMPs and TIMPs in pancreatic carcinoma, normal pancreas, and pancreatic carcinoma cell lines has been determined by Northern analysis. The transcripts have been localized by in situ hybridization and the MMP2 protein by immunohistochemistry. Expression of MMP2, 7, and -11 was greater in pancreatic carcinoma than in normal pancreas (P < 0.01). MMP7 expression in normal pancreas and MMP7 and -11 expression in tumours was always seen the TIMP1 expression. TIMP2 was expressed in only half of the tumours and a previously undescribed transcript size is reported for TIMP2. MTMMP was expressed concurrently with MMP2 in 64 per cent of tumours, but concurrent MMP2 and TIMP2 expression occurred in only half. MMP2 mRNA was found more often in the tumour stroma than with the other MMPs or TIMPs (P < 0.02). It is concluded that while overexpression of MMP7 and -11 may be countered by TIMP1, the aggressive phenotype of pancreatic carcinoma may occur because of overexpression of MMP2, activated by MTMMP and associated with a reduced expression of TIMP2. PMID- 9349240 TI - Metastatic lesions from prostate cancer do not express oestrogen and progesterone receptors. AB - Oestrogen receptors (ER) and progesterone receptors (PR) have been reported by several authors in the stromal cells of the human prostate. Controversial results exist on the expression of ER and PR in epithelial cells of the prostate. Some recent publications, in contrast to previous findings, have suggested that these receptors are also present in human prostate cancer cell lines derived from metastatic lesions. The expression of ER and PR in these cell lines has been re examined to determine their presence in lymph node metastases from patients who did not receive any kind of endocrine therapy and in distant metastases obtained from patients who failed endocrine treatment. ER and PR expression in LNCaP, PC 3, and DU-145 cells was assessed by means of the reverse transcriptase-polymerase chain reaction, ligand binding assays, and immunohistochemistry. With all the techniques applied, the three cell lines were found to be negative for both ER and PR. Immunohistochemical analyses were performed in four lymph node metastases obtained at radical prostatectomy from patients who did not receive endocrine therapy and in 17 distant metastases obtained at palliative surgery from patients who failed endocrine therapy. All 21 metastases were negative for ER and PR on immunohistochemistry. These results do not support the recently developed concept that receptors for oestrogenic and progestagenic steroids are present in metastases from human prostate cancer. PMID- 9349241 TI - Induction of myocardial heat shock protein 70 during cardiac surgery. AB - Animal experiments have shown that members of the heat shock protein (HSP) family have cytoprotective properties against ischaemia. In experimentally induced cardiac ischaemia, the induction of HSP70s correlates with reduced infarct size and enhanced myocardial function and endothelial recovery. Direct evidence that increased myocardial HSP70 expression result in cytoprotection during ischaemia has also been obtained using transgenic mice overexpressing either rat or human HSP72. This study examined the induction and expression of myocardial HSP70s after an obligatory period of ischaemia in patients during cardiac surgery. The level of HSP72/HSC73 protein in Tru-cut biopsies of the myocardium, taken before and after an acute ischaemic insult, was examined using a polyclonal antibody. The amount of HSP72 mRNA in the biopsies was also determined by reverse transcriptase polymerase chain reaction (RT-PCR) and correlated HSP72/HSC73 protein expression. In four patients subjected to brief alternating periods of normothermic ischaemia and reperfusion, the amount of myocardial HSP72/HSC73 protein was increased several fold after ischaemic insult. This was accompanied by increased expression of HSP72 mRNA. In contrast, the amounts of myocardial HSP72/HSC73 protein and HSP72 mRNA were unchanged in a patient subjected to a single prolonged period of hypothermic ischaemia. Given the proven myocardial protective properties of HSP72 in experimental models, it is postulated that the observed induction of HSP72 may have a similar function in man. PMID- 9349242 TI - Labelling pattern obtained by non-isotopic in situ hybridization as a prognostic factor in HPV-associated lesions. PMID- 9349243 TI - Where is efficiency leading our system of health insurance? PMID- 9349244 TI - What do managed care plans do to affect care? Results from a survey of physicians. AB - Little is known about physicians' exposure to managed care techniques that affect clinical practice. In 1995, we conducted a survey of 2,003 U.S. physicians asking them about their share of patients subject to a variety of managed care techniques. Nationally, 24% of physicians received some form of capitation payment for their patients. The two most widely used techniques were utilization review (UR), applied to an average of 59% of patients, and discounted fees, applied to an average of 38% of patients. Although UR was common, ultimate denial rates of coverage were very low: at most 3% for the types of care studied. Use of managed care techniques varied more within states than between states. Conventional measures of HMO market penetration revealed little about how managed care affects physicians. PMID- 9349245 TI - The impact of HMO penetration on the rate of hospital cost inflation, 1985-1993. AB - This paper provides evidence that growth in health maintenance organization (HMO) enrollment slows hospital cost inflation. During the period 1985-1993, hospitals in areas with high rates of HMO penetration and growth had a slower rate of growth in expenses (8.3%) than hospitals in low penetration areas (11.2%). From 1992-1993, HMO growth lowered the rate of hospital cost inflation by .34 to 3.40 percentage points, depending on the base-year level and the annual change in HMO penetration. Declines in Medicare Prospective Payment System (PPS) margins also lowered hospital cost inflation; over the time period, annual hospital cost inflation was reduced by .38 percentage points. The estimates imply that the cumulative effect of HMO growth on hospital costs has been a $56.2 billion reduction (in 1993 dollars). PMID- 9349246 TI - Association of HMO penetration and other credit quality factors with tax-exempt bond yields. AB - This paper evaluates the relationship of HMO penetration, as well as other credit quality measures of market, institutional, operational, and financial traits, to tax-exempt bond yields. The study analyzed more than 1,500 bond issues from 1990 through 1993 and corrected for simultaneous relationships between bond size and yield and selection bias. The study found lower bond yields for hospitals located in markets with no HMO penetration. Lower yields for bond issues also were found for facilities generating higher numbers of days cash on hand and greater debt service coverage. Finally, results show that hospitals with higher occupancy rates achieve a lower yield. PMID- 9349247 TI - Structural incentives and adoption of medical technologies in HMO and fee-for service health insurance plans. AB - Recent literature has argued that conventional fee-for-service (FFS) health insurance, as compared to managed care (HMO) insurance, may lead to the adoption of new technology that raises costs and reduces patient welfare. In this paper, we show that this result depends on an increasingly unrealistic key assumption that FFS insurers cannot refuse to reimburse new technology. We also show that, when the assumption is changed, HMO insurers may adopt costly technologies that FFS insurers do not. PMID- 9349248 TI - Switching to managed care in the small employer market. AB - In 1993, only 22% of small employers offered a managed care product; by 1995, nearly 70% did. This study uses nationally representative data on small firms in 1993 and 1995 to examine the factors underlying this dramatic shift. Two explanations emerge from the regression work. Adoption of managed care by large employers appears to have served as a signal, certifying the acceptance of managed care among workers. Second, lower prices for managed care products, relative to conventional insurance, increased the adoption of managed care, particularly in 1995. There is little evidence that state insurance reforms prompted the switch, although they may have helped set the stage for it. PMID- 9349249 TI - Small group reform in a competitive managed care market: the case of California, 1993 to 1995. AB - State-level insurance reforms designed to make health insurance more accessible for small businesses and their employees have become common in the 1990s. This study examines the effects of small group reform legislation enacted in California in 1993. Using survey data on health benefits in small firms, we look at changes in health insurance coverage that occurred between spring 1993 (just before reform) and spring 1995. Our results indicate that insurance became slightly more affordable and, among businesses with three to nine employees, employer provision increased more than 10 percentage points. Provision was unchanged among larger-sized businesses, however. Managed care penetration increased considerably. We argue that California's competitive health insurance market, which already was dominated by managed care, represented a favorable environment for small group reform. In this context, the modest growth in insurance provision highlights the limited potential of incremental reforms for expanding insurance coverage. PMID- 9349250 TI - Cloning of two plant cDNAs encoding a beta-type proteasome subunit and a transformer-2-like SR-related protein: early induction of the corresponding genes in tobacco cells treated with cryptogein. AB - We report the successful combination of mRNA differential-display reverse transcription PCR (DDRT-PCR) and 5'-rapid amplification of cDNA ends (5'-RACE) in order to isolate full-length cDNAs corresponding to genes activated in tobacco cells treated with cryptogein within 60 min. Cloning and sequencing of two cDNAs, called 'tcI 7' and 'tcI 14' (for tobacco cryptogein-induced), allowed the identification of open reading frames. Deduced amino-acid sequences of 'tcI 7' and 'tcI 14' showed significant homologies with a beta-type proteasome subunit and a transformer-2-like serine/arginine-rich (SR) ribonucleoprotein, respectively. The accumulation of mRNAs corresponding to 'tcI 7' started 30 min after the addition of cryptogein to tobacco cell suspensions and continued up to 180 min, whereas the accumulation of 'tcI 14' corresponding mRNAs was transitory between 30 and 60 min. These results indicated a transcriptional activation of the corresponding genes early after elicitation of tobacco cells by cryptogein. The biological significance of this activation remains to be elucidated. PMID- 9349251 TI - Isolation and characterization of cDNA for a plant mitochondrial phosphate translocator (Mpt1): ozone stress induces Mpt1 mRNA accumulation in birch (Betula pendula Roth). AB - We have isolated by DDRT-PCR (differential-display reverse-transcription polymerase chain reaction) and cDNA library screening a 1.3 kb cDNA corresponding to a strongly ozone-inducible transcript from birch (Betula pendula Roth). Nucleotide sequence analysis suggests that it encodes a mitochondrial phosphate translocator protein (Pic), the first one isolated from plants. The isolated birch mitochondrial phosphate translocator cDNA (designated Mpt1) contains an open reading frame of 1092 bases encoding a 364 amino acid polypeptide. The deduced protein is 66% similar to bovine Pic isoform B. Comparison of the N terminal amino acid sequence to known mammalian Pic proteins and the existence of an in-frame stop codon upstream of the initiation codon suggest that the isolated cDNA is full-length. Southern hybridization analysis of birch genomic DNA shows that Mpt1 is a single-copy gene. Accumulation of Mpt1 mRNA during oxidative stress imposed by ozone is detectable already at 2 h and it is at maximum ca. 12 h after the beginning of an 8 h ozone exposure (150 ppb). A second O3 peak at 48 56 h did not increase transcript levels further. O3 exposure for 2 h was sufficient for Mpt1 induction. Birch Mpt1 transcript levels remain at moderately low level during leaf development and is lower in roots and leaves when compared to young shoots undergoing wood formation and lignification. PMID- 9349253 TI - Correct blue-light regulation of pea Lhcb genes in an Arabidopsis background. AB - Irradiation of etiolated Arabidopsis or pea, or dim-red-light-grown pea seedling with a single, short (under 10 s) pulse of blue light (threshold at 0.1 mumol/m2) is sufficient to induce the expression of specific members of the Lhcb gene family including the pea Lhcb1*4 gene and the Arabidopsis Lhcb1*3 gene. Other Lhcb genes, such as the pea Lhcb1*3 gene and the Arabidopsis Lhcb1*1 and 1*2 genes are unaffected by this blue-light treatment. Transgenic Arabidopsis bearing pea Lhcb1*3::Gus (beta-glucuronidase), pea Lhcb1*4::Gus or Arabidopsis Lhcb1*3::Gus constructs were used to determine if pea and Arabidopsis employ a similar mechanism to achieve blue-light induced Lhcb expression. Examination of the respective Gus expression patterns in white-light-grown seedlings indicates that the pea promoters are active and properly expressed in the Arabidopsis background. Irradiation of dark-grown Arabidopsis with a 20 s pulse of blue light with a total fluence of 100 mumol/m-2 results in expression of the pea Lhcb1*4::Gus (beta-glucuronidase) construct, but not of the pea Lhcb1*3::Gus construct indicating that the pea promoters respond correctly to blue light in the Arabidopsis background. Fluence-response, time-course and reciprocity characteristics for the blue-light-induced expression of the pea Lhcb1*4::Gus construct closely resemble those of the endogenous Arabidopsis Lhcb genes, confirming the proper interpretation of the Arabidopsis blue-light-signaling mechanism by the pea Lhcb1*4 promoter and suggesting that the signaling mechanisms in the two plants are very similar, if not identical. Fluence response data for the steady-state level of transcript derived from an Arabidopsis Lhcb1*3::Gus construct extending 200 bp upstream of the site of transcription indicate that the blue light responsive elements(s) are contained within this 200 bp region. PMID- 9349252 TI - Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter. AB - A study of the expression of a bean phenylalanine ammonia-lyase (PAL) promoter/beta-glucuronidase gene fusion in transgenic tobacco has shown that the PAL2 promoter has a modular organization. Expression of the PAL2 promoter in the vascular system involves positive and negative regulatory cis elements. Among these elements is an AC-rich motif implicated in xylem expression and a suppressing cis element for phloem expression. Using radiolabelled complementary oligonucleotides bearing the AC-rich motif, a cDNA clone encoding a DNA-binding protein has been isolated from a tobacco lambda gt11 expression library. This factor, named AC-rich binding factor (ACBF), showed binding specificity to the AC rich region. The specificity of ACBF for the AC-rich region was also shown using a gel retardation assay with an ACBF recombinant protein extract. The deduced amino acid sequence from ACBF contains a long repeat of glutamine residues as found in well characterized transcription factors. Interestingly, ACBF shared sequence similarity to conserved amino acid motifs found in RNA-binding proteins. Genomic gel blot analysis indicated the presence of a small gene family of sequences related to ACBF within the tobacco nuclear genome. Analysis of tobacco mRNA using the ACBF cDNA as probe showed that while ACBF mRNA was present in all tissues examined, the highest transcript accumulation occurred in stem tissues. The functional characteristics of the AC-rich sequence were examined in transgenic tobacco. A heptamer of the AC-rich sequence, in front of a minimal 35S promoter from cauliflower mosaic virus (-46 to +4), conferred specific expression in xylem. PMID- 9349254 TI - Molecular cloning and expression analysis of dihydroflavonol 4-reductase gene in flower organs of Forsythia x intermedia. AB - The expression, during flower development, of the gene encoding the anthocyanin pathway key enzyme dihydroflavonol 4-reductase (DFR) was investigated in floral organs of Forsythia x intermedia cv. 'Spring Glory'. Full-length DFR and partial chalcone synthase (CHS) cDNAs, the gene of interest and a flavonoid pathway control gene respectively, were obtained from petal RNA by reverse transcription PCR. Whereas for CHS northern blot analysis enabled the study of its expression pattern, competitive PCR assays were necessary to quantify DFR mRNA levels in wild-type plants and in petals of 2 transgenic clones containing a CaMV 35S promoter-driven DFR gene of Antirrhinum majus. Results indicated a peak of CHS and DFR transcript levels in petals at the very early stages of anthesis, and different expression patterns in anthers and sepals. In comparison to wild-type plants, transformants showed a more intense anthocyanin pigmentation of some vegetative organs, and a dramatic increase in DFR transcript concentration and enzymatic activity in petals. However, petals of transformed plants did not accumulate any anthocyanins. These results indicate that other genes and/or regulatory factors should be considered responsible for the lack of anthocyanin production in Forsythia petals. PMID- 9349255 TI - Analysis of multiple copies of geminiviral DNA in the genome of four closely related Nicotiana species suggest a unique integration event. AB - Previously, we discovered multiple direct repeats of geminivirus-related DNA (GRD) sequences clustered at a single chromosomal position in Nicotiana tabacum (tobacco). Here we show that, in addition to tobacco, multiple copies of these elements occur in the genomes of three related Nicotiana species, all in the section Tomentosae: N. tomentosiformis, N. tomentosa and N. kawakamii, but not in 9 other more distantly related Nicotiana species, nor in various other solanaceous and non-solanacous plants. DNA sequence analysis of 18 GRD copies reveal 4 distinct, but highly related, sub-families: GRD5, GRD3 and GRD53 in tobacco; GRD5 in N. tomentosiformis and N. kawakamii; and GRD2 in N. tomentosa. In addition to novel sequences, all elements share significant but varying lengths of DNA sequence similarity with the geminiviral replication origin plus the adjacent rep gene. There is extended sequence similarity to REP protein at the deduced amino acid sequence level, including motifs associated with other rolling circle replication proteins. Our data suggest that all GRD elements descend from a unique geminiviral integration event, most likely in a common ancestor of these Tomentosae species. PMID- 9349257 TI - Geranylgeranyl pyrophosphate synthase encoded by the newly isolated gene GGPS6 from Arabidopsis thaliana is localized in mitochondria. AB - We have cloned a new geranylgeranyl pyrophosphate (GGPP) synthase gene, designated GGPS6, from Arabidopsis thaliana genomic DNA. Nucleotide sequence analysis revealed that the GGPS6 gene contains an open reading frame coding for a protein of 343 amino acid residues with a calculated molecular mass of 37,507 Da. Also, the gene is not interrupted by an intron. The predicted amino acid sequence of the GGPS6 gene shows significant homology (34.0-57.7%) with other GGPP synthases from Arabidopsis. The GGPS6 protein contains a N-terminal signal peptide which is thought to function as an organelle targeting sequence. In fact, the GGPS6-GFP fusion protein was found to be localized exclusively in mitochondria when expressed in tobacco BY-2 cells. In vitro analysis of the enzyme activity as well as genetic complementation analysis with Erwinia uredovora crt gene cluster expressed in Escherichia coli showed that the GGPS6 gene most certainly encodes a GGPP synthase catalyzing the conversion of farnesyl pyrophosphate to GGPP. PMID- 9349256 TI - Expression of a synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures. AB - A synthetic antifreeze protein gene was expressed in plants and reduced electrolyte leakage from the leaves at freezing temperatures. The synthetic AFP was expressed as a fusion to a signal peptide, directing it to the extracytoplasmic space where ice crystallization first occurs. The gene was introduced to Solanum tuberosum L. cv. Russet Burbank by Agrobacterium-mediated transformation. Transformants were identified by PCR screening and expression of the introduced protein was verified by immunoblot. Electrolyte-release analysis of transgenic plant leaves established a correlation between the level of transgenic protein expression and degree of tolerance to freezing. This is the first identification of a phenotype associated with antifreeze protein expression in plant tissue. PMID- 9349258 TI - Aerobic fermentation during tobacco pollen development. AB - In vegetative organs of plants, the metabolic switch from respiration to fermentation is dictated by oxygen availability. The two genes dedicated to ethanolic fermentation, pyruvate decarboxylase and alcohol dehydrogenase, are induced by oxygen deprivation and the gene products are active under oxygen stress. In pollen, these two genes are expressed in a stage-specific manner and transcripts accumulate to high levels, irrespective of oxygen availability. We have examined the expression pattern of pyruvate decarboxylase and alcohol dehydrogenase at the protein level in developing pollen and show that the active proteins are localized to the gametophytic tissue and begin to accumulate at microspore mitosis. A flux through the ethanolic fermentation pathway could already be detected very early in pollen development, occurring in all stages from premeiotic buds to mature pollen. This flux was primarily controlled not by oxygen availability, but rather by sugar supply. At a high rate of sugar metabolism, respiration and fermentation took place concurrently in developing and germinating pollen. We propose that aerobic fermentation provides a shunt from pyruvate to acetyl-CoA to accommodate the increased demand for energy and biosynthetic intermediates during pollen development and germination. A possible undesirable side-effect is the potential accumulation of toxic acetaldehyde. Our results support a model for cms-T-type male sterility in maize, in which degeneration of the tapetum is caused by the toxic effects of acetaldehyde on mitochondria weakened by the presence of the URF13 protein. PMID- 9349259 TI - Aldehyde dehydrogenase in tobacco pollen. AB - Acetaldehyde is one of the intermediate products of ethanolic fermentation, which can be reduced to ethanol by alcohol dehydrogenase (ADH). Alternatively, acetaldehyde can be oxidized to acetate by aldehyde dehydrogenase (ALDH) and subsequently converted to acetyl-CoA by acetyl-CoA synthetase (ACS). To study the expression of ALDHs in plants we isolated and characterized a cDNA coding for a putative mitochondrial ALDH (TobAldh2A) in Nicotiana tabacum. TobALDH2A shows 54 60% identity at the amino acid level with other ALDHs and shows 76% identity with maize Rf2, a gene involved in restoration of male fertility in cms-T maize. TobAldh2A transcripts and protein were present at high levels in the male and female reproductive tissues. Expression in vegetative tissues was much lower and no induction by anaerobic incubation was observed. This suggests that TobALDH expression is not part of the anaerobic response, but may have another function. The use of specific inhibitors of ALDH and the pyruvate dehydrogenase (PDH) complex indicates that ALDH activity is important for pollen tube growth, and thus may have a function in biosynthesis or energy production. PMID- 9349260 TI - Zrp2: a novel maize gene whose mRNA accumulates in the root cortex and mature stems. AB - A near full-length cDNA clone (pZRP2) was isolated from a cDNA library constructed from maize root mRNAs. The predicted polypeptide has a calculated molecular mass of 66,975 Da, is largely hydrophilic, and contains 26 repeats of a motif the consensus sequence of which is RKATTSYG[S][D/E][D/E][D/E][D/E][P]. The function of the putative protein remains to be elucidated. The ZRP2 mRNA accumulates to the highest levels in young roots, and is also present in mature roots and stems of maize. Further analysis of young roots indicates that the lowest level of ZRP2 mRNA is near the root tip, with relatively high levels throughout the remainder of the root. In situ hybridization reveals that ZRP2 mRNA accumulates predominantly in the cortical parenchyma cells of the root. In vitro nuclear run-on transcription experiments indicate a dramatically higher level of zrp2 gene transcription in 3-day old roots than in 5-day old leaves. A zrp2 genomic clone, which includes the transcribed region and 4.7 kb of upstream sequence, was isolated and characterized. PMID- 9349261 TI - Expression of chalcone synthase and chalcone isomerase proteins in Arabidopsis seedlings. AB - Antibodies have been developed against the first two enzymes of flavonoid biosynthesis in Arabidopsis thaliana. Chalcone synthase (CHS) and chalcone isomerase (CHI) were overexpressed and purified from Escherichia coli as fusion proteins with glutathione S-transferase from Schistosoma japonicum. The recombinant proteins were then used to immunize chickens and the resulting IgY fraction was purified from egg yolks. Immunoblots of crude protein extracts from Arabidopsis seedlings carrying wild-type and null alleles for CHS and CHI showed that the resulting antibody preparations provide useful tools for characterizing expression of the flavonoid pathway at the protein level. An initial analysis of expression patterns in seedlings shows that CHS and CHI proteins are present at high levels during a brief period of early seedling germination that just precedes the transient accumulation of flavonoid end-products. PMID- 9349262 TI - Rare codons are not sufficient to destabilize a reporter gene transcript in tobacco. AB - In plants, as in other eukaryotes, most synonymous codons of the genetic-code are not used with equal frequency, but instead some codons are preferred, whereas others are rare. Circumstantial evidence led to the suggestion that rare codons have a negative influence on mRNA stability. To address this question experimentally, rare codons encoded by a Bacillus thuringiensis (B.t.) toxin gene (cryIA(c)) or a synthetic sequence were introduced into a phytohemagglutinin (PHA) reporter gene. In neither case was the mRNA stability appreciably diminished in stably transformed tobacco cell cultures nor was the accumulation of mRNA in transgenic plants affected. Thus rare codons do not appear to be sufficient to cause rapid degradation of the PHA mRNA and potentially other mRNAs in plants. PMID- 9349264 TI - Lumenal proteins involved in respiratory electron transport in the cyanobacterium Synechocystis sp. PCC6803. AB - Cyanobacterial thylakoids catalyze both photosynthetic and respiratory activities. In a photosystem I-less Synechocystis sp. PCC 6803 strain, electrons generated by photosystem II appear to be utilized by cytochrome oxidase. To identify the lumenal electron carriers (plastocyanin and/or cytochromes c553, c550, and possibly cM) that are involved in transfer of photosystem II-generated electrons to the terminal oxidase, deletion constructs for genes coding for these components were introduced into a photosystem I-less Synechocystis sp. PCC 6803 strain, and electron flow out of photosystem II was monitored in resulting strains through chlorophyll fluorescence yields. Loss of cytochrome c553 or plastocyanin, but not of cytochrome c550, decreased the rate of electron flow out of photosystem II. Surprisingly, cytochrome cM could not be deleted in a photosystem I-less background strain, and also a double-deletion mutant lacking both plastocyanin and cytochrome c553 could not be obtained. Cytochrome cM has some homology with the cytochrome c-binding regions of the cytochrome Caa3-type cytochrome oxidase from Bacillus spp. and Thermus thermophilus. We suggest that cytochrome cM is a component of cytochrome oxidase in cyanobacteria that serves as redox intermediate between soluble electron carriers and the cytochrome aa3 complex, and that either plastocyanin or cytochrome c553 can shuttle electrons from the cytochrome b6f complex to cytochrome cM. PMID- 9349263 TI - Characterization of Phaseolus vulgaris cDNA clones responsive to water deficit: identification of a novel late embryogenesis abundant-like protein. AB - Six cDNA clones from Phaseolus vulgaris, whose expression is induced by water deficit and ABA treatment (rsP cDNAs) were identified and characterized. The sequence analyses of the isolated clones suggest that they encode two types of late-embryogenesis abundant (LEA) proteins, a class-1 cytoplasmic low-molecular weight heat shock protein (lmw-HSP), a lipid transfer protein (LTP), and two different proline-rich proteins (PRP). One of the putative LEA proteins identified corresponds to a novel 9.3 kDa LEA-like protein. During the plant response to a mild water deficit (psi w = -0.35 MPa) all genes identified present a maximal expression at around 16 or 24 h of treatment, followed by a decline in expression levels. Rehydration experiments revealed that those genes encoding PRPs and LTP transiently re-induce or maintain their expression when water is added to the soil after a dehydration period. This is not the case for the lea genes whose transcripts rapidly decrease, reaching basal levels a few hours after rehydration (4 h). Under water deficit and ABA treatments, the highest levels of expression for most of the genes occur in the root, excluding the ltp gene whose maximum expression levels are found in the aerial regions of the plant. This indicates that for these genes, both water deficit and ABA-dependent expression are under organ-specific control. The data presented here support the importance of these proteins during the plant response to water deficit. PMID- 9349265 TI - Regulated expression of plant tRNA genes by the prokaryotic tet and lac repressors. AB - The prokaryotic tet operator (tetO) sequence was inserted at positions upstream and downstream of sequences encoding the Arabidopsis thaliana tRNA(Lys)AUC or tRNA(Trp)AUC suppressor tRNAs, and tRNA expression in carrot protoplasts was measured by translational suppression of a nonsense codon in a luciferase reporter gene. Regulation of tRNA expression by the tetracycline repressor (tetR) occurred from genes with the tetO inserted at position -1 (for the tRNA(Trp)AUC gene), or at positions -2, -6 and -10 (for the tRNA(Lys)AUC gene), and repression reached 90%. The inducer tetracycline (Tc) restored tRNA expression. Similarly, carrot protoplasts transfected with human tRNA(Ser)AUC genes containing the lac operator (lacO) in their 5'-flanking sequence with or without the lac repressor (lacI) gene, conditionally expressed tRNAs which suppressed the luciferase reporter. Up to 30-fold repression occurred by the lactose repressor when lacO was located at position -1 of the tRNA(Ser)AUC coding sequence. In the presence of the inducer isopropyl-beta-thiogalactoside (IPTG), repression was relieved. These results demonstrate that sequences flanking tRNA genes can strongly influence tRNA expression in plants, and in a conditional fashion when bound by inducible proteins. PMID- 9349266 TI - A promoter directing high level expression in pistils of transgenic plants. AB - The promoter of the potato (Solanum tuberosum L.) SK2 gene, encoding a pistil specific basic endochitinase, was cloned. Various fragments of the SK2-promoter, from 1 kb down to 0.23 kb in length, were fused to the GUS reporter gene. Chimaeric SK2 promoter-GUS fusion constructs were transformed into potato by Agrobacterium tumefaciens-mediated transformation. The SK2-GUS transgenic potato plants exhibited a highly specific GUS activity in the pistil. Expression in the pistil was shown to be developmentally regulated. In addition to the GUS activity in pistils, transgenic plants also showed a much weaker ectopic expression in anthers. In other tissues no systematic expression was detectable. All SK2 promoter fragments analysed conferred pistil-specific expression without significant qualitative or quantitative differences, demonstrating that the regulatory elements mediating this expression pattern are located within a 230 bp SK2 promoter fragment. The SK2 promoter may be used to engineer high levels of expression in pistils of transgenic plants. PMID- 9349267 TI - Two growth-related organ-specific cDNAs from Cicer arietinum epicotyls. AB - Two cDNAs, CanST-1 and CanST-2, encoding two different growth-related organ specific sequences have been isolated from a cDNA library from growing epicotyls of Cicer arietinum. An intriguing property of these two clones is the presence in their coding region of a repeated sequence which is highly conserved except for the number of repeats. The corresponding genes of CanST-1 and CanST-2 encode for proteins related to elongation processes. CanST-1 and CanST-2 are up-regulated during epicotyl growth, the transcript levels of both clones decrease when the growth of epicotyls is inhibited by several treatments and their expression increases when epicotyls resume growth. Furthermore, clones CanST-1 and CanST-2 are tissue-specific and are only expressed in epicotyls, mesocotyls, roots and stem tissues whose cells undergo elongation processes. Neither clone was found to be expressed in other organs such as cotyledons, leaves, flowers, pods and immature seeds. The results of auxin (IAA) and brassinolides (BR) treatments suggest that the processes in which the proteins encoded by CanST-1 and CanST-2 are involved are not mediated by these hormones. PMID- 9349268 TI - Expression of an Antirrhinum dihydroflavonol reductase gene results in changes in condensed tannin structure and accumulation in root cultures of Lotus corniculatus (bird's foot trefoil). AB - Condensed tannins (proanthocyanidins) are an important factor in the nutritive and dietary quality of many forage crops. We report here experiments aimed at altering the levels and monomer composition of condensed tannins (CTs) in 'hairy root' cultures of Lotus corniculatus (bird's foot trefoil) using genetic manipulation. An Antirrhinum majus dihydroflavonol reductase (DFR) cDNA was expressed in sense in L. corniculatus and CT levels in transgenic root cultures were analysed. Two co-transformed lines were noted with decreased CT content relative to controls and these levels were comparable with antisense-DFR phenotypes. In ADFR10, a co-transformed line with the highest CT levels, CT structure was altered in a manner consistent with the substrate specificity of the introduced gene; that is an increase in pro-pelargonidin monomers noted after hydrolysis of CTs. RT-PCR confirmed the expression of endogenous DFR gene(s) in both putatively co-suppressed lines and also in ADFR10. Analysis of selected root culture lines indicated that the monomer composition of CTs did not change during growth and development but that levels of CTs varied in a regulated manner. PMID- 9349269 TI - cDNA sequence, genomic organization and differential expression of three Arabidopsis genes for germin/oxalate oxidase-like proteins. AB - Wheat germin is a protein expressed during germination which possesses an oxalate oxidase activity. Germin-type oxalate oxidases have been extensively studied in monocotyledons (wheat and barley) where they are thought to have important functions for development, stress response and defence against pathogens. In contrast, almost nothing is known about the germin-like proteins found in dicotyledons, gymnosperms and myxomycetes. In this work, cDNA clones for three genes (ATGER1, ATGER2 and ATGER3) encoding germin-like proteins, initially characterized as expressed sequence tags (ESTs), from Arabidopsis thaliana cDNA libraries were further characterized. In addition, we isolated and sequenced a Brassica napus cDNA which was strongly homologous to the cDNA for ATGER1. Sequence analysis and secondary structure predictions of the proteins encoded by these cDNAs showed that they possess all the characteristic features of members of the germin family and of the germin/seed globulins/sucrose binding protein superfamily. Sequence comparisons and mapping demonstrated the existence of at least two different gene families in the A. thaliana genome encoding a minimum of three genes for germins. These three genes have been mapped in three different location on the Arabidopsis genome. By northern blot hybridizations we found that these genes are differentially regulated. ATGER1 was expressed during germination, like wheat germin, but also in leaves whereas ATGER2 transcripts were exclusively found in developing embryos, like wheat pseudo-germin. ATGER3 mRNAs were found in leaves and flowers and their abundance was shown to vary during the circadian cycle. PMID- 9349270 TI - Molecular characterization of glyoxalase II from Arabidopsis thaliana. AB - Glyoxalase II is part of the glutathione-dependent glyoxalase detoxification system. In addition to its role in the detoxification of cytotoxic 2-oxo aldehydes, specifically methylglyoxal, it has been suggested that the glyoxalase system may also play a role in controlling cell differentiation and proliferation. During the analysis of a T-DNA-tagged mutant of Arabidopsis we identified the gene for a glyoxalase II isozyme (GLY1) that appears to be mitochondrially localized. The cDNA encoding a glyoxalase II cytoplasmic isozyme (GLY2) was also isolated and characterized. Southern blot and sequence analyses indicate that glyoxalase II proteins are encoded by at least two multigene families in Arabidopsis. Escherichia coli cells expressing either GLY1 or GLY2 exhibit increased glyoxalase II activity, confirming that they do, in fact, encode glyoxalase II proteins. Northern analysis shows that the two genes are differentially expressed. Transcripts for the mitochondrial isozyme are most abundant in roots, while those for the cytoplasmic isozyme are highest in flower buds. The identification of glyoxalase II isozymes that are differentially expressed suggests that they may play different roles in the cell. PMID- 9349271 TI - Cloning and characterization of rac-like cDNAs from Arabidopsis thaliana. AB - The Rho family of GTPases are in higher eukaryotes divided into 3 major subfamilies; the Rho, Rac and Cdc42 proteins. In plants, however, the Rho family is restricted to one large family of Rac-like proteins. From work with mammalian phagocytes the Rac proteins are known to activate a multicomponent NADPH dependent oxidase which results in accumulation of H2O2, a process termed oxidative burst. In plants a similar oxidative burst is observed and plays and important role in its defence against pathogen infections, suggesting a similar role for the plant Rac-like proteins. The Rho family of GTPases proteins are also involved in control of cell morphology, and are also thought to mediate signals from cell membrane receptors. In a broad search for members of the Ras superfamily in plants, several new small GTP-binding proteins were found. We report here the identification and molecular cloning of 5 rac-like cDNAs from Arabidopsis thaliana, Arac1-5. The Rac-like proteins deduced from the cDNA sequences all share 80-95% homology, but show considerably more diversity on the nucleotide level, indicating that this is an ancient gene family. Four of the rac genes were found to be expressed in all tissues examined, but one gene, Arac2, was expressed exclusively in the root, hypocotyl and stem. Our results show that the rac gene family in A. thaliana consists of at least 10 different genes. PMID- 9349273 TI - Differential modification of flavonoid and isoflavonoid biosynthesis with an antisense chalcone synthase construct in transgenic Lotus corniculatus. AB - Three clonal genotypes of Lotus corniculatus L. (bird's foot trefoil) were transformed with an antisense chalcone synthase (CHS) gene construct made using a stress induced CHS17 cDNA from Phaseolus vulgaris under the control of the constitutive CaMV 35S promoter and Nos terminator via Agrobacterium rhizogenes. After initial screening, ten antisense and five control co-transformation events from each recipient clonal genotype were analysed. After elicitation with glutathione, the level of tannin accumulation was found to be increased in a number of antisense root cultures derived from the low (S33) and moderate (S50) tannin recipient genotypes. Six antisense and four control transformed lines from genotype S50 were selected for more detailed study. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from 1 to 5 and antisense orientation was confirmed by PCR. In transformed root cultures, increased CHS transcript levels were noted in a number of antisense lines. Biochemical analyses of glutathione-elicited-root cultures indicated a significant increase in tannin accumulation in antisense CHS lines and mean vestitol levels were reduced. These results show that the introduction of a heterologous antisense chalcone synthase construct into L. corniculatus resulted in an unpredicted molecular and biochemical phenotype. Such findings are discussed in relation to manipulation of this complex multigene family. PMID- 9349272 TI - Two genes encoding extension-like proteins are predominantly expressed in tomato root hair cells. AB - A differential screen of a tomato root hair cDNA library resulted in the cloning of two cDNAs, Dif10 and Dif54, whose corresponding genes are preferentially expressed in root hair cells as determined by analysis of mRNA levels in various tomato organs. Transcript levels showed no increase in leaves subjected to hormonal and environmental stress treatments. Sequence analysis of the cDNAs revealed high similarity to members of the extension family. Extensions are hydroxyproline-rich glycoproteins (HRGPs) located in the cell wall. In order to study the functional significance of HRGPs in root hairs, tomato seedling roots were treated with micromolar concentrations of 3,4-dehydro-L-proline (Dhp), a selective inhibitor of prolyl hydroxylase. Dhp treatment resulted in changes in root growth and the development of root hairs with reduced hair length, suggesting an important role of HRGPs in hair morphogenesis. PMID- 9349275 TI - Isolation, characterization and molecular cloning of a leaf-specific lectin from ramsons (Allium ursinum L.). AB - Lectins were isolated from roots and leaves of ramsons and compared to the previously described bulb lectins. Biochemical analyses indicated that the root lectins AUAIr and AUAIIr are identical to the bulb lectins AUAI and AUAII, whereas the leaf lectin AUAL has no counterpart in the bulbs. cDNA cloning confirmed that the leaf lectin differs from the bulb lectins. Northern blot analysis further indicated that the leaf lectin is tissue-specifically expressed. Sequence comparisons revealed that the ramsons leaf lectin differs considerably from the leaf lectins of garlic, leek, onion and shallot. PMID- 9349274 TI - Gene targeting approaches using positive-negative selection and large flanking regions. AB - We report here on strategies aimed at improving the frequency of detectable recombination in plants by increasing the efficiency of selecting double recombinants in transgenic calli. Gene targeting was approached on the Gln1 and the Pzfloci of Lotus japonicus, using Agrobacterium tumefaciens T-DNA replacement vectors. Large flanking regions, up to 22.9 kb, surrounding a positive selection marker were presented as substrates for homologous recombination. For easier detection of putative recombinants the negative selectable marker cytosine deaminase was inserted at the outside borders of the flanking regions offered for cross-over. A combination of positive and negative selection allowing double recombinants to grow, while counter-selecting random insertions, was used to select putative targeting events. The more than 1000-fold enrichment observed with replacement vectors designed to minimize gene silencing demonstrated the efficiency of the negative selection. Using five different replacement vectors an estimated total of 18,974 transformation events were taken through the positive negative selection procedure and 185 resistant calli obtained. Targeting events could not be verified in the survivors by PCR screening and Southern blot analysis. With this approach the frequency of detectable gene targeting in L. japonicus was below 5.3 x 10(-5), despite the large flanking sequences offered for recombination. PMID- 9349276 TI - Isolation and characterization of an Arabidopsis biotin carboxylase gene and its promoter. AB - In the plastids of most plants, acetyl-CoA carboxylase (ACCase; EC 6.4.1.2) is a multisubunit complex consisting of biotin carboxylase (BC), biotin-carboxyl carrier protien (BCCP), and carboxytransferase (alpha-CT, beta-CT) subunits. To better understand the regulation of this enzyme, we have isolated and sequenced a BC genomic clone from Arabidopsis and partially characterized its promoter. Fifteen introns were identified. The deduced amino acid sequence of the mature BC protein is highly conserved between Arabidopsis and tobacco (92.6% identity). BC expression was evaluated using northern blots and BC/GUS fusion constructs in transgenic Arabidopsis. GUS activity in the BC/GUS transgenics as well as transcript level of the native gene were both found to be higher in silique and flower than in root and leaf. Analysis of tobacco suspension cells transformed with truncated BC promoter/GUS gene fusions indicated the region from -140 to +147 contained necessary promoter elements which supported basal gene expression. A positive regulatory region was found to be located between -2100 and -140, whereas a negative element was possibly located in the first intron. In addition, several conserved regulatory elements were identified in the BC promoter. Surprisingly, although BC is a low-abundance protein, the expression of BC/GUS fusion constructs was similar to 35S/GUS constructs. PMID- 9349277 TI - Isolation of the cDNAs encoding (+)6a-hydroxymaackiain 3-O-methyltransferase, the terminal step for the synthesis of the phytoalexin pisatin in Pisum sativum. AB - Pisatin is the major phytoalexin produced by pea upon microbial infection. The enzyme that catalyzes the terminal step in the pisatin biosynthetic pathway is (+)6a-hydroxymaackiain 3-O-methyltransferase (HMM). We report here the isolation and characterization of two HMM cDNA clones (pHMM1 and pHMM2) made from RNA obtained from Nectria haematococca-infected pea tissue. The two clones were confirmed to encode HMM activity by heterologous expression in Escherichia coli. The substrate specificity of the methyltransferases in E. coli was similar to the activity detected in CuCl2-treated pea tissue. Nucleotide sequence analysis of Hmm1 and Hmm2 revealed an open reading frame of 1080 bp and 360 amino acid residues which would encode 40.36 kda and 40.41 kDa polypeptides, respectively. The deduced amino acid sequence of HMM1 has 95.8% identity to HMM2, 40.6% identity to Zrp4, a putative O-methyltransferase (OMT) in maize root, and 39.1% to pBH72-F1, a putative OMT induced in barley by fungal pathogens or UV light. Comparison of the deduced amino acid sequences of the cDNA clones to OMTs from other higher plants identified the binding sites of S-adenosylmethionine (AdoMet). Southern blot analysis showed two closely linked genes with strong homology to Hmm in the pea genome. PMID- 9349278 TI - Developmental, stress and ABA modulation of mRNA levels for bZip transcription factors and Vp1 in barley embryos and embryo-derived suspension cultures. AB - The transcription factors VP1 (Viviparous-1), EmBP-1 (Em-binding protein 1) and OSBZ8, originally cloned and analysed in various monocot species, have been implicated in the regulation of the Lea (late embryogenesis-abundant) group 1 genes. We have investigated the modulation of the levels of these mRNAs in barley during embryogenesis, and in developing embryos subjected to various kinds of osmotic stress. The accumulation of mRNA for VP1 and EmBP-1 transcription factors, using cDNAs cloned from barley, starts at 10 and 15 days after anthesis, respectively, whereas Lea B19 mRNA levels are very low or undetectable until 25 days after anthesis during normal development. The EmBP-1 mRNA is predominantly induced in mannitol-stressed immature embryos. Vp1 mRNA was not significantly modulated by ABA, salt or mannitol. Inhibition of ABA biosynthesis by norflurazon showed that the induction of both Vp1 and EmBP-1 mRNAs was ABA-independent. In embryo-derived suspension-cultured cells, neither of the two transcripts would be induced by ABA or osmotic stress, although both OSBZ8 and one member of the Lea B19 family was up-regulated by ABA. Electrophoretic mobility shift assays using a Lea B19.1 probe with an ABRE (abscisic acid-responsive element) similar to that which binds EmBP-1 and OSBZ8 in the wheat and rice Em promoters show that the binding activity is increased by ABA and osmotic stress. Taken together, these data show that both VP1 and EmBP-1 are involved in embryo-specific signal transduction pathways, that they are differentially regulated at the mRNA level, and that EmBP-1 can be induced by osmotic stress independently of any increase in endogenous ABA. The difference in mRNA regulation patterns of OSBZ8 and EmBP-1 may suggest that they are involved in different signal transduction pathways in connection with osmotic stress/ABA regulation of Lea genes. PMID- 9349279 TI - Eucalyptus has functional equivalents of the Arabidopsis AP1 gene. AB - Two Eucalyptus homologues of the Arabidopsis floral homeotic gene AP1 (EAP1 and EAP2) show 60-65% homology to AP1. EAP1 and EAP2 are expressed predominantly in flower buds. EAP2 produces two different polypeptides arising from differential splicing at an intron, the shorter EAP2 protein diverging from the longer sequence after amino acid 197 and having a translation stop after residue 206. This truncated protein includes both MADS- and K-box amino acid sequences. Ectopic expression of the EAP1 or either of the two EAP2 polypeptides in Arabidopsis driven by the 35S promoter produces effects similar to the corresponding AP1 construct, causing plants to flower earlier, have shorter bolts and resemble the terminal flower mutant (tfl). PMID- 9349281 TI - UDP-glucose:sterol glucosyltransferase: cloning and functional expression in Escherichia coli. AB - Steryl glucosides are characteristic lipids of plant membranes. The biosynthesis of these lipids is catalyzed by the membrane-bound UDP-glucose:sterol glucosyltransferase (EC 2.4.1.173). The purified enzyme (Warnecke and Heinz, Plant Physiol 105 (1994): 1067-1073) has been used for the cloning of a corresponding cDNA from oat (Avena sativa L.). Amino acid sequences derived from the amino terminus of the purified protein and from peptides of a trypsin digestion were used to construct oligonucleotide primers for polymerase chain reaction experiments. Screening of oat and Arabidopsis cDNA libraries with amplified labeled DNA fragments resulted in the isolation of sterol glucosyltransferase-specific cDNAs with insert lengths of ca. 2.3 kb for both plants. These cDNAs encode polypeptides of 608 (oat) and 637 (Arabidopsis) amino acid residues with molecular masses of 66 kDa and 69 kDa, respectively. The first amino acid of the purified oat protein corresponds to the amino acid 133 of the deduced polypeptide. The absence of these N-terminal amino acids reduces the molecular mass to 52 kDa, which is similar to the apparent molecular mass of 56 kDa determined for the purified protein. Different fragments of these cDNAs were expressed in Escherichia coli. Enzyme assays with homogenates of the transformed cells exhibited sterol glucosyltransferase activity. PMID- 9349280 TI - Characterization of the gene family for alternative oxidase from Arabidopsis thaliana. AB - We investigated the copy number of the gene for alternative oxidase (AOX) of Arabidopsis thaliana by amplification by PCR and Southern hybridization. These studies indicated that there are at least four copies of the AOX gene in Arabidopsis. We isolated genomic clones containing individual copies (designated as AOX1a, AOX1b, AOX1c and AOX2) of the AOX genes. Interestingly, two of the AOX genes (AOX1a and AOX1b) were located in tandem in a ca. 5 kb region on one of the chromosomes of Arabidopsis. Comparison between genomic and cDNA sequences of the four AOX genes showed that all AOX genes are divided by three introns and the positions of the introns in AOX1a, AOX1b, AOX1c and AOX2 are the same. We examined whether expression of Arabidopsis AOX genes, like the tobacco AOX1a gene, is enhanced by treatment with antimycin A, an inhibitor of complex III in the mitochondrial respiratory chain. We found that, in young plants, the amount of Arabidopsis AOX1a mRNA was dramatically increased by addition of antimycin A, while the transcription of the other three genes (AOX1b, AOX1c and AOX2) did not respond to antimycin A. Amplification by RT-PCR showed that AOX1a and AOX1c were expressed in all organs examined (flowers and buds, stems, rosette, and roots of 8-week old plants). In contrast, transcripts of AOX1b were detected only in the flowers and buds, and transcripts of AOX2 were detected mainly in stems, rosette and roots. These results suggested that transcriptions of the four genes for alternative oxidase of Arabidopsis are differentially regulated. PMID- 9349282 TI - Cloning and expression of the pea gene encoding SBP65, a seed-specific biotinylated protein. AB - Besides biotin-dependent carboxylases, which play key roles in basic metabolism, SBP65 (seed biotinylated protein of 65 kDa of apparent molecular mass), an atypical biotinylated protein, has been described in pea plants. This seed specific protein is devoid of any carboxylase activity, and shares many physiological and molecular features with late embryogenesis-abundant (Lea) proteins. In a first step toward understanding the role of this peculiar protein, we have demonstrated the role of abscisic acid (ABA) and of the osmotic environment on its expression using northern blot analysis from immature embryos cultured in vitro and germinating mature seeds. Moreover, the cloning and characterization of its gene (referred to as sbp gene) allowed us to define various potential cis-acting elements within the promoter region to account for the observed strict seed-specific expression. The results described in this paper are consistent with a model in which ABA regulates, at least in part, expression of this gene. However, unlike most lea genes, ABA regulation of the sbp gene seems to occur in a very restricted fashion, being confined only to particular stages of embryo development. Such a strict spatial and temporal expression pattern is dependent on the osmotic environment of the developing embryos and on tissue-specific factors, presumably preventing biotin depletion in cells requiring this essential cofactor for basic metabolic activity. PMID- 9349283 TI - Heteromeric assembly of the cytosolic glutamine synthetase polypeptides of Medicago truncatula: complementation of a glnA Escherichia coli mutant with a plant domain-swapped enzyme. AB - We have cloned and sequenced the cDNAs corresponding to the two cytosolic glutamine synthetase (GS) polypeptides (a and b) of Medicago truncatula. Using these two cDNAs we have prepared a construct encoding the N-terminal domain of b and the C-terminal domain of a in order to produce a domain-swapped polypeptide which should assemble to give an enzyme containing chimeric active sites. Both the native and the domain-swapped enzymes were expressed in Escherichia coli where they were catalytically and physiologically active as they were able to rescue a glnA deletion mutant. The expressed polypeptides were of the correct size and the isoenzymes behaved similarly to their native homologues on ion exchange chromatography. We have found slight differences in the kinetic properties of the purified enzymes and in the modulation of their activities by several putative cellular effectors. In vitro dissociation of the purified a and b homo-octamers, followed by reassociation, showed that the subunits are able to self-assemble, perhaps randomly, to form heteromeric isoenzymes. Moreover, heteromeric isoenzymes occur in the plant as revealed by studies on the GS isoenzymes of nodules, roots, stems and stipules. PMID- 9349284 TI - Expression of glyoxylate cycle genes in cucumber roots responds to sugar supply and can be activated by shading or defoliation of the shoot. AB - When cucumber roots are excised and incubated without a carbon source, isocitrate lyase (ICL) and malate synthase (MS) mRNAs increase significantly in amount. However, if sucrose is added to the excised roots, the mRNAs do not accumulate. Hairy roots obtained by transformation with Agrobacterium rhizogenes show the same response. Transgenic hairy roots containing the Icl and Ms gene promoters fused to the GUS reporter gene, have very low GUS activity which increases dramatically when roots are incubated in the absence of sugar, indicating regulation at the transcriptional level. Staining of sugar-deprived roots shows that GUS activity is concentrated mainly in root tips and lateral root primordia, where demand for carbohydrate is greatest. In order to determine if Icl and Ms genes are expressed in roots of whole plants under conditions which may occur in nature, cucumber plants were subjected to reduced light intensity or defoliation. In both cases increases were observed in ICL and MS mRNAs. These treatments also reduced root sugar content, consistent with the hypothesis that sugar supply could control expression of Icl and Ms genes in roots of whole plants. PMID- 9349285 TI - Cloning and characterisation of a cytosolic glutathione reductase cDNA from pea (Pisum sativum L.) and its expression in response to stress. AB - A second glutathione reductase (GR) cDNA has been cloned and sequenced from pea (Pisum sativum L. cv. Birte). This new GR cDNA (GOR2) does not encode a pre protein with a transit peptide and therefore is most likely to represent a cytosolic GR. It is significantly different at the DNA level from the previously cloned chloroplastidial/mitochondrial pea GR (GOR1), but retains the features characteristic of GRs from all sources and has GR activity when expressed in Escherichia coli. GOR2 maps to linkage group 6 on the pea genome map and it seems likely that this is the only locus for this gene. In contrast to GOR1, transcript levels of GOR2 increase in the recovery (post-stress) phases of both drought and chilling by about ten- and three-fold respectively. GOR2 therefore may play a role in the restoration of the post-stress redox state of the cytosolic glutathione pool. PMID- 9349286 TI - Structural analysis of rDNA in the genus Nicotiana. AB - The nucleotide sequence of the intergenic spacer (IGS) region between the 25S and the 18S rRNA coding regions has been determined for tobacco (Nicotiana tabacum). The IGS (5140 bp in length) can be subdivided into several regions (I-VII) two of which, upstream and downstream of the putative transcription initiation site (TIS), contain prominent subrepeats (A and C). The unique sequence in the central part of the IGS (region IV) preceding the TIS is extremely AT-rich. The distance from the putative TIS to the 5' end of the 18S rRNA gene is 3005 bp. The IGS sequences are compared with potato (Solanum tuberosum) and tomato (Lycopersicon esculentum) IGS. Restriction mapping of 13 Nicotiana species shows that considerable rDNA repeat length heterogeneity in this genus is probably due to different numbers of A and C subrepeats. PMID- 9349287 TI - Induction of a Citrus gene highly homologous to plant and yeast thi genes involved in thiamine biosynthesis during natural and ethylene-induced fruit maturation. AB - Maturing citrus fruit undergo pigment changes which can be enhanced by exogenous ethylene. In order to identify genes induced by ethylene in citrus fruit peel, we cloned the gene c-thi1. mRNA corresponding to c-thi1 increased gradually in the peel during natural fruit maturation and in response to ethylene. GA3 pretreatment reduced the inductive effect of ethylene. Levels of c-thi1 increased also in juice sacs but the effect of ethylene was much less prominent. c-thi1 is homologous to yeast and plant genes encoding for an enzyme belonging to the pathway of thiamine biosynthesis. The data suggest that thiamine is involved in citrus fruit maturation. PMID- 9349288 TI - Cloning of upstream sequences responsible for cell cycle regulation of the Nicotiana sylvestris CycB1;1 gene. AB - To understand the mechanisms involved in the regulation of the mitotic cyclin B Nicta; CycB1;1 expression, we have cloned the Nicotiana sylvestris cyclin gene, Nicsy; CycB1;1, whose coding sequence is homologous to that of Nicta;CycB1;1 cDNA. The structure and the function of its 5'-flanking region, 1149 bp upstream of the first start codon, was analysed. By producing stably transformed cells of a synchronized culture with the Nicsy;CycB1;1 promoter/beta-glucuronidase (gus) reporter gene fusion, we demonstrate that the 1149 bp promoter fragment mediates a gus transcriptional oscillation, indistinguishable from that of endogenous Nicsy;CycB1;1 cyclin B transcripts. Transient GUS activity in BY-2 protoplasts reveals that promoter activity is considerably reduced by shortening the 5' flanking region to 538 or 320 bp. Furthermore, the 320 bp fragment no longer mediates the observed transcriptional regulation of the 1149 bp Nicsy;CycB1;1 promoter in BY-2 protoplasts isolated from synchronized cells. PMID- 9349289 TI - In vivo characterization of transcriptional regulatory sequences involved in the defence-associated expression of the tobacco retrotransposon Tnt1. AB - The expression of the tobacco retrotransposon Tnt1 is induced by wounding, pathogen infections as well as microbial elicitors and abiotic factors known to induce the plant defence response. We report here that the LTR U3 region is sufficient to mediate transcriptional activation by biotic and abiotic elicitors in stable transgenic conditions. We have used in vivo footprinting techniques in order to analyse the cis-regulatory elements of the LTR U3 region that mediate the induction of Tnt1 expression. Our results indicate that a tandemly repeated short element, named BII box, is involved in the transcriptional activation of the tobacco retrotransposon Tnt1 in association with the plant defence signaling cascade. PMID- 9349290 TI - Multiple hepatitis virus infections in chronic HBsAg carriers in Naples. AB - In order to determine the prevalence of multiple infections with hepatitis viruses in chronic HBsAg carriers in Naples, to assess the interaction between HBV, HDV and HCV infections and to evaluate the influence of multiple virus hepatitis infections on the clinical presentation, we studied 198 HBsAg chronic carriers observed consecutively from 1971 to 1988 at our Liver Unit. Of the 198 HBsAg chronic carriers, 171 had undergone percutaneous liver biopsy. The presence of HBcAg or HDAg in the liver biopsy was considered a marker of HBV or HDV replication, respectively; the presence of anti-HCV was considered a marker of HCV infection. Anti-HCV was observed in 13.6% of the 22 subjects with normal liver, in 27.7% of the 47 patients with minimal chronic hepatitis, in 40% of the 50 with mild chronic hepatitis, in 70.6% of the 17 with moderate hepatitis, in 66.7% of the 3 with severe chronic hepatitis and in 65.6% of the 32 with active cirrhosis. Anti-HCV positive cases were antiHD positive more frequently than the anti-HCV negative (59.2% vs. 43%, p = 0.05). HDV infection exerted a clear inhibition on the HBV genome. Among the 171 HBsAg chronic carriers, the finding of an active chronic hepatitis (moderate chronic hepatitis + severe chronic hepatitis + active cirrhosis) is less frequent in subjects with HBV replication alone than in those with HDV replication or HCV infection. Patients with both HBV replication and HCV infection and those with both HDV replication and HCV infection showed a very high prevelance of active chronic hepatitis. PMID- 9349291 TI - Mother to infant transmission of coinfection by human immunodeficiency virus and hepatitis C virus: prevalence and clinical manifestations. AB - The prevalence and the clinical course of hepatitis C virus (HCV) infections were studied in 23 HIV-1-infected children, who were born to 22 mothers with HIV-1/HCV coinfection. During the follow-up only two children (8.7%) showed persistent anti HCV antibodies and circulating HCV RNA. Both children, who were aged 10 and 10.6 years respectively at the end of follow-up, had chronically-evolving liver disease and autoimmune thrombocytopenia but no signs of progressive HIV disease. Based on our experience, vertically-acquired HIV-1/HCV coinfection is less frequent than is generally reported and may be associated with the development of chronic thrombocytopenia in addition to liver disease. Moreover, perinatal HIV 1/HCV coinfection appears to be associated with a slow progression of HIV disease. PMID- 9349292 TI - Quantification and genotyping of serum HCV-RNA in patients with chronic hepatitis C undergoing interferon treatment. AB - Quantification of serum HCV-RNA and HCV genotyping was studied in 27 patients with chronic hepatitis C undergoing interferon treatment. Pretreatment serum HCV RNA levels were quantified using competitive RT-PCR and compared to a quantitative RT-PCR assay based on co-amplification of HCV-RNA with a synthetic RNA standard. HCV genotyping was performed using a line probe reversed hybridisation assay or direct solid-phase sequencing. This study shows the feasibility of performing HCV-RNA quantification. RT-PCR based on co amplification HCV-RNA titer less than 6 x 10(4) genome equivalents/ml serum did correlate with a complete sustained response to alpha interferon in chronic hepatitis C. HCV genotype 1b was alpha predominantly associated with a high non responder rate. Future prospective trials will be required to evaluate quantitative HCV-RNA levels and HCV genotyping as response predicting parameters for interferon-treatment. PMID- 9349293 TI - Quantitative analysis of hepatitis C virus activity in vivo in different groups of untreated patients. AB - Highly sensitive competitive PCR (cPCR) and competitive reverse transcription PCR (cRT-PCR) methodologies were recently developed and applied for quantifying viral DNA and RNA species (including HCV RNA) present in clinical samples at low concentration. In this study, we used cRT-PCR to compare the viral load of 118 untreated patients with HCV infection and different clinical conditions (80 patients with chronic hepatitis, 18 infected subjects with persistently normal ALT levels and various degrees of liver injury, 10 HCV infected subjects that tested positive for anti-LKM1 antibodies, and 10 patients with HCV infection and cryoglobulinemia). The results indicate that while great individual variability of HCV viremia is detectable even among patients with similar clinical conditions, the mean HCV RNA copy number in samples from patients with different clinical conditions was similar in all groups with the single exception of patients that tested positive for anti-liver-kidney microsomal auto-antibodies type 1 (anti-LKM1); interestingly, lower HCV viremia levels were revealed in these anti-LKM1-positive cases with liver disease of uncertain pathogenesis. PMID- 9349294 TI - Non-specific and specific anti-HCV results correlated to age, sex, transaminase, rhesus blood group and follow-up in blood donors. AB - Second generation enzyme immunoassays (EIA-2) for antibodies to hepatitis C virus (anti-HCV) have a higher specificity and sensitivity than first generation enzyme immunoassays (EIA-1). We studied how many anti-HCV-positive blood donors were missed by the EIA-1, how many were false positive, how false-positive donors should be dealt with and how the results of the EIA-2 correlate to demographic data and serum alanine aminotransferase (ALT) level. A total of 208, 544 northern German blood donors, not preselected for anti-HCV negativity, were tested for anti-HCV with EIA-2 and, if repeatability reactive (rr), were retested with a licensed supplementary test (RIBA-2). 0.43% of the donors were EIA-2 rr, but only 0.12% of women and 0.09% of men were RIBA-2 positive. RIBA-2 positivity rates were very low in donors 18 to 27 years old (0.03% and 0.05%) and rose with age in women but not in men. Infected women were significantly more often Rhesus negative than men. The rate of unspecifically positive EIA-2 results (entirely negative in RIBA-2) increased with age in both sexes and did not correlate with ALT. The ALT distribution was age-dependent with a different pattern for men and women. Confirmation of EIA-2 results with RIBA was rare when ALT was low and frequent when ALT was high. ALT screening before introduction of Anti-HCV detected one out of six infected donors. To exclude this one infectious donation, 46 uninfected donations had to be excluded in addition. Only 8% of the then RIBA 2-positive donors were not detected by EIA-1. Apparent seroconversions in EIA-2 are usually not specific; only 1 out of 66 apparent seroconversions could be confirmed by RIBA-2 suggesting recent HCV infection. 0.15% of the donor population showed an inconsistent EIA-2 pattern during follow-up. We conclude that donors should not be excluded from further donations, even on the basis of multiple EIA-1 positive results or on the basis of only one EIA-2 positive donation. Anti-D-immunoglobulin prophylaxis may have been a source of infection in some Rhesus-negative women. ALT screening should not be discontinued because recent HCV infection can be detected earlier by ALT than by anti-HCV, but exclusion limits should be elevated to increase specificity and limit unnecessary exclusion of donations. PMID- 9349295 TI - Receptor-mediated entry of hepatitis B virus particles into liver cells. AB - In previous reports several receptors for either natural hepatitis B virus (HBV) particles or genetically engineered virus have been described, whereby endocytosis represents a putative uptake mechanism for HBV particles. We have found that HBV-particles from viremic carriers could bind to the human asialoglycoprotein receptor (ASGPR), which mediates glycoprotein uptake into liver cells. The HBV-ASGPR interaction was studied in a cell culture system using hepatoma HepG2 and HuH7 cells compared to COS cells as controls. About 50% of HBsAg-secretion into the cell culture supernatant after HBV-inoculation as a function of HBV-uptake could be inhibited by the specific ASGPR-ligand asialofetuin. COS-cells did not show HBsAg-secretion. If the cells were grown as clones, 15% of HepG2-cells demonstrated HBsAg-secretion but only 5% in the presence of asialofetuin. HBV-particle uptake was further confirmed by HBV-DNA analysis using PCR. HBV-ASGPR interaction was studied with purified, biotin conjugated human ASGPR. Quantitative inhibition with asialofetuin indicated a high-affinity binding of HBV-particles to purified ASGPR. After denaturing polyacrylamid gel electrophoresis and transblotting of isolated HBV-particles a receptor-blotting system was established which identified distinct binding sites for biotinylated receptors. These results suggest that the ASGPR is capable of specifically binding HBV-particles and, moreover, to mediate their hepatic endocytosis which ultimately could be responsible for the HBV-infection of liver cells. PMID- 9349296 TI - Woodchuck hepatitis virus DNA integration in a common chromosomal region of the woodchuck genome in two independent hepatocellular carcinomas. AB - In woodchuck hepatocellular carcinoma (HCC) the myc-oncogene family (particularly N-myc2) and the win locus of cellular genome have been reported as frequent targets for integration of woodchuck hepatitis virus (WHV) DNA. In this paper a further cellular locus, b3n, is reported as recurrent target for WHV integration in woodchuck HCC. Cloning and sequencing of a WHV-DNA integration and its cellular flanking regions showed that viral DNA was inserted in a chromosomal region already described for WHV integration in another single HCC. The two integration sites are only 0.5 kb apart. A link between WHV integration in b3n and HCC development may be postulated. Careful analysis of the sequence of the unoccupied locus revealed that, in addition to Alu-like repeats and a gag-like coding region, already described, several features of Matrix Attachment Region (MAR) sequences are present. Thus (part of) b3n might be a previously unrecognized MAR. Organization of the chromatin in functional domains and regulation of gene expression are some functions attributed to MAR sequences. The occurrence of WHV-DNA integration close to the same putative MAR in two different HCCs suggests that a mechanism of deregulation of MAR functions by WHV insertion might act in some liver tumors. PMID- 9349297 TI - Determination of peripheral blood mononuclear cell responses to mitogens and woodchuck hepatitis virus core antigen in woodchucks by 5-bromo-2'-deoxyuridine or 2[3H]adenine incorporation. AB - Characterization of cellular immune response to antigens of woodchuck hepatitis virus (WHV) can contribute to the understanding of acute resolving and chronic outcome of hepadnavirus infection. Studies were limited because peripheral blood mononuclear cells (PBMCs) of woodchucks failed to incorporate [3H]thymidine sufficiently. Therefore, we established a non-radioactive proliferation assay for woodchuck PBMCs using 5-bromo-2'-deoxyuridine (BrdU) as thymidine analogue. Mitogen- and WHV core protein-(WHcAg) induced PBMC proliferation was detected by BrdU incorporation and compared to an assay using 2[3H]adenine. After stimulation with concanavalin A (ConA) and phytohaemagglutinin (PHA) we observed significant PBMC proliferation with both assays. Mitogen-induced nucleoside uptake of PBMCs into cellular DNA was confirmed by detection of 1',2'[3H]BrdU and 2[3H]adenine in extracted DNA. PBMCs obtained during the acute phase of WHV infection could be stimulated by WHcAg, whereas no WHcAg-induced proliferation of PBMCs was found in WHV-negative animals. PBMCs of chronic WHV carriers showed only a weak response to WHcAg. The established assays will be useful in determining the kinetics of cellular immune responses to different WHV antigens in the course of WHV infection and may provide an insight into mechanisms responsible for chronic outcome of hepadnavirus infection. PMID- 9349298 TI - Characterization of antibody response to hepatitis C virus protein E2 and significance of hypervariable region 1-specific antibodies in viral neutralization. AB - Antibodies directed against hypervariable region 1 (HVR1) within the viral glycoprotein E2 of hepatitis C virus (HCV) are postulated to neutralize virus. An in vitro infection/binding assay of human fibroblast cells was established in order to study neutralization of HCV. Occurrence of mutations in the nucleotide sequence of HVR1 as compared to the inoculum after infection of human fibroblasts suggested replication of HCV in these cells. The significance of HVR1-specific antibodies in sera of patients who were infected in a single-source outbreak by an HCV contaminated anti-D immunoglobulin (IgG) preparation was studied. Using immunoprecipitation and ELISA, HVR1-specific antibodies could be detected in most of the sera obtained early (< or = 1 year p.i.) and late (up to 14 years p.i.) in single patients. Further characterization of the HVR1-specific antibodies in patient sera by attachment studies of HCV to the human fibroblasts suggested that HVR1-specific antibodies in sera obtained early p.i. can neutralize virus of the anti-D IgG preparation. PMID- 9349299 TI - Impaired response to alpha interferon in patients with an inapparent hepatitis B and hepatitis C virus coinfection. AB - The possibility of hepatitis B virus (HBV) infection in HBsAg-negative patients has been shown. However, an "inapparent" coinfection by HBV in hepatitis C virus (HCV)-positive patients generally is not taken into account in clinical practice. Mechanisms responsible for resistance to interferon (IFN) have not been completely clarified. The aim of this study was to investigate whether an "inapparent" coinfection by HBV in anti-HCV-positive chronic liver disease patients may influence IFN response. Fourteen anti-HCV positive, HBsAg-negative but serum HBV DNA-positive patients by PCR and 111 anti-HCV-positive, HBsAg negative and HBV DNA (PCR)-negative patients with chronic hepatitis were treated with 3 MU of recombinant alpha-2a IFN 3 times weekly for 12 months. Serum HBV DNA and HCV RNA were determined before treatment, after 6-12 months and in coincidence with ALT flare-up by PCR. HBV PCR was performed using primers specific for the S region of the HBV genome and HCV PCR with primers localised in the 5'NC region of HCV genome. IgM anti-HBc was tested using IMx Core-M Abbott assay. By the end of treatment, ALT values had become normal in 4/14 HBV DNA positive patients (28%), but all "responders" (4/4) relapsed between 2 and 5 months after therapy. All but one patient were HCV RNA-positive before treatment, 6 were also both HBV DNA and HCV RNA-positive during ALT flare-ups. In 5 patients, only HBV DNA and in 3 patients, only HCV RNA was detected when transaminase values increased. All patients remained HBsAg-negative and anti-HCV positive. IgM anti-HBc was detected both before treatment and during ALT elevation in 3 patients and only during ALT relapse in 3 others. Of the 111 anti HCV positive, HBsAg-negative and HBV DNA (PCR)-negative patients with chronic hepatitis, a biochemical response to IFN treatment was observed in 54% of the cases. Relapse of ALT values was observed in 47% of the cases during a follow-up of 1 year after treatment. "Inapparent" HBV/HCV coinfection may be implicated in cases of resistance to IFN treatment. In addition, HBV replication may persist in patients in whom HCV replication was inhibited by IFN treatment. The pathogenic role of HBV in liver disease was confirmed by detection of IgM anti-HBc in some cases; the appearance of these antibodies only after IFN treatment suggests that IFN may exert a selective role in favour of HBV. Further studies will show the effect of different treatment schedules. HBV DNA and/or IgM anti-HBc detection with very sensitive methods may be important both as a prognostic factor and as a tool for better understanding interviral relationships and mechanisms involved in multiple hepatitis virus infections. PMID- 9349300 TI - Hepatitis C virus infection in mixed cryoglobulinemia and B-cell non-Hodgkin's lymphoma: evidence for a pathogenetic role. AB - We investigated the pathogenetic relevance of hepatitis C virus (HCV) infection in mixed cryoglobulinemia (MC) with or without complicating B-cell Non-Hodgkin's lymphoma (NHL) in comparison with other immunological and lymphoproliferative disorders. The following groups of patients were studied: A) 25 patients with MC in 7 cases evolved into B-cell NHL; B) 25 healthy subjects; C) 22 patients with different systemic immune diseases; D) 24 patients with chronic HCV infection without MC; E) 25 patients with B-cell idiopathic NHL. Methods used included: i) Polymerase chain reaction (PCR) for HCV RNA detection in serum and peripheral blood mononuclear cells (PBMC) (uncultured or mitogen-stimulated); ii) Branched DNA (b-DNA) for HCV RNA quantification; iii) HCV genotyping by genotype-specific primers localized in the core region and by hybridization of amplification products of the 5' untranslated region (5'UTR), obtained with universal primers, using genotype-specific probes. Serum anti-HCV and HCV RNA were detected in 88% and 73% of MC patients, respectively, and in a significantly lower percentage of healthy controls and patients with autoimmune diseases. HCV RNA concentration was significantly lower in supernatants than in corresponding whole sera (p < 0.001). Plus-strand HCV RNA was detected in 81% of peripheral blood mononuclear cell (PBMC) samples and minus-strand in the majority of fresh or mitogen stimulated cells. All MC patients with NHL had HCV RNA sequences in PBMC. HCV genotype 2a/III was detected in MC patients with a prevalence that was significantly higher than in HCV infected patients without MC. Surprisingly, HCV markers (anti HCV and/or HCV RNA) were found in 32% of patients with idiopathic NHL. These data suggest that HCV infection is involved in the pathogenesis of MC through both direct participation in the immune complex related vasculitis and by triggering the lymphoproliferative disorder underlying the disease. This latter disorder seems to be related to HCV lymphotropism which could also be responsible for the evolution of MC to malignant lymphoma. This study also suggests that HCV infection may be involved in the pathogenesis of idiopathic B-cell NHL through a similar pathogenetic mechanism. PMID- 9349301 TI - HCV infection of peripheral blood mono nuclear cells and serum levels of soluble ICAM-1 in patients treated with interferon. AB - Serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) have been examined in 38 patients with chronic hepatitis C liver disease treated with interferon. The sICAM-1 values were found to correlate significantly with the ALT values. Pre-treatment sICAM-1 values of responder and nonresponder patients were not significantly different while, by the end of the treatment, the values of responders were significantly lower compared to those of nonresponders. However, no difference could be found between sustained and relapse responders. Of the 21 patients examined for PBMC HCV-RNA, 15 (71.4%) were found to be positive. Neither the rate of responsivity to interferon treatment, nor the mean sICAM-1 values correlated with the positivity of PBMC HCV-RNA. However, the clearance of serum and PBMC HCV-RNA was associated to a significant decrease of sICAM-1 and ALT levels. In conclusion, sICAM-1 values were found to correlate with ongoing viral replication and liver cytonecrosis, but were not influenced by the concomitant HCV infection of PBMC. PMID- 9349302 TI - Soluble intercellular adhesion molecule-1 in sera from patients with chronic hepatitis C undergoing interferon treatment. AB - To study the influence of interferon therapy on soluble intercellular adhesion molecule-1 we measured sICAM-1 levels in 22 patients with type C chronic hepatitis treated with interferon. We also studied 9 healthy subjects as control group. The results showed statistically significant higher levels of sICAM-1 in patients with liver disease than in the controls. The sICAM-1 baseline levels were similar in patients with chronic active hepatitis or cirrhosis but, during therapy, these levels decreased only in patients with chronic hepatitis. After IFN withdrawal sICAM-1 levels rebounded to initial values. PMID- 9349303 TI - Ganciclovir therapy for cytomegalovirus-associated liver disease in immunocompetent or immunocompromised children. AB - Ganciclovir therapy was given intravenously to 20 children with cytomegalovirus (CMV)-associated liver disease, of whom 6 were immunocompetent and 14 were immunocompromised (9 had AIDS and 5 had solid tumors). Immunocompetent children had isolated liver disease diagnosed at birth (4 children), or systemic congenital CMV infection including liver disease (2 children). Ganciclovir was used following two regimens: A) 5 mg/kg twice daily for 8 to 86 days (mean 21); B) 7.5 mg/kg twice daily for 14 days followed by 10 mg/kg three times weekly for three months. CMV infection was diagnosed by viral isolation, detection of viral antigens, and/or CMV DNA from blood and urine. All immunocompetent children had negative CMV culture and CMV DNA detection from blood and/or urine after 14 weeks of treatment. However, the three children who were treated with regimen B showed normal ALT levels at the end of the maintenance course, whereas the children who received ganciclovir with regimen A had normal ALT levels only after about 1 year. All children with tumors initiated regimen B, but only three, who had negative CMV detection and markedly decreased ALT levels, received full treatment; of the remaining two children, one recovered after only an initial course, and the other had therapy interrupted because of hepatic failure and died 9 days later. In contrast, the children with AIDS received several ganciclovir courses for different periods at the lower dosage: they generally improved during treatment but did not recover completely, and five children died with active CMV infections. Based on our study, CMV-associated liver disease can be efficiently treated with ganciclovir both in immunocompetent and immunodeficient children. However, a single ganciclovir course including a higher dosage and prolonged therapy appeared to be more effective than several courses with lower dosages. PMID- 9349304 TI - Different susceptibilities of genetic variants of hepatitis C virus (HCV) to interferon (IFN). AB - Genetic variants of HCV may have different degrees of resistance to IFN and may therefore influence the outcome of IFN therapy. However, selection of HCV variants by IFN has not been investigated in detail. In this paper, heteroduplex analysis was used to monitor major changes of HCV populations in 4 chronically infected patients under IFN therapy. We found that a major variant of the HCV 5' non-coding region (5' NCR) emerged in a responder. In other patients although no new variant of the 5' NCR was identified, significant changes occurred within the core and E1 region of the HCV genome. Disappearance and emergence of HCV variants may reflect their different susceptibilities to IFN. Our results indicate that responses of HCV populations to IFN are complex and need to be characterized by analysis of multiple HCV genome regions. PMID- 9349305 TI - Core specific antisense phosphorothioate oligodeoxynucleotides as potent and specific inhibitors of hepatitis C viral translation. AB - Antisense phosphorothioate oligodeoxynucleotides (S-ODN) complementary to sequence stretches in the 5' non coding region (NCR) of the hepatitis C virus (HCV) have recently been shown to effectively inhibit viral gene expression. In order to further delineate the optimum target region in the highly conserved 5' end of the viral RNA, S-ODN complementary to HCV core coding sequences were analysed in the present study. In a rabbit reticulocyte lysate (RRL) in vitro translation assay S-ODN 5, complementary to the HCV-RNA nucleotides 340-353, and S-ODN-6, complementary to nucleotides 348-365, resulted in an inhibition of viral translation of 90.4 +/- 1.3% and 93.7 +/- 5.1%, respectively at a concentration of 4.14 microM. S-ODN 7, complementary to nucleotides 371-388, was relatively inefficient and showed a maximal inhibition of 42.4 +/- 12.2%. It has been suggested that in living cells an inhibition by S-ODN is mainly mediated by the action of RNAse H. In RRL the RNAseH content is very low; therefore, to simulate the situation in living cells inhibition experiments in RRL enriched with RNAse H were performed. Under these conditions S-ODN 5, 6 and 7 inhibited viral translation by 45.6 +/- 6.3%, 80.3 +/- 2.8% and 70.9 +/- 5.7% at concentrations as low as 0.2 microM. At this concentration no inhibition was observed in the standard RRL assay. In cell culture S-ODN 7 was by far the most efficient inhibitor of viral translation, resulting in a specific inhibition of 89.4 +/- 3.6% at a concentration of 0.3 microM. Taken together with the results of our previous study, nucleotides 326-348 comprising the 3' end of the NCR and nucleotides 371-388, located entirely in the core coding region of the HCV RNA, are effective targets for S-ODN mediated inhibition of viral translation. PMID- 9349306 TI - Recombinant human antibodies specific for hepatitis C virus proteins. AB - Human antibodies to hepatitis C virus core, NS4A and NS3 were cloned in a prokaryotic vector and expressed as soluble Fab fragments and as phage-displayed Fabs. The recombinant Fabs were shown to be a suitable tool for immunohistochemistry, since they recognize the cognate antigen expressed in mammalian cells. The nucleotide sequence of the cDNA for the variable domains of these antibodies was determined and the V-gene usage was derived. On the basis of the deduced amino acid sequence, a structural model of the V domains of the Fabs was constructed. PMID- 9349307 TI - Novel molecular approaches toward therapy of chronic hepatitis B. AB - More than 300 million people worldwide are chronically infected with hepatitis B virus (HBV), and are at greatly increased risk of developing liver cirrhosis and eventually primary liver carcinoma. While infection can, with relative success, be prevented by vaccination, no generally effective therapy for chronic hepatitis B is available. Hence there is an urgent need for novel antiviral strategies. Recent advances in our understanding of the mechanisms underlying virus replication and assembly provide opportunities for the rational design of molecules that could specifically interfere with these processes; some of these possibilities are discussed in this review. PMID- 9349308 TI - Nidovirales: a new order comprising Coronaviridae and Arteriviridae. PMID- 9349309 TI - Clinical pathway for peritoneal carcinomatosis from colon and rectal cancer: guidelines for current practice. AB - Peritoneal carcinomatosis from colon and rectal cancer was in the past a terminal condition without curative treatment options. Recently, an aggressive treatment strategy has resulted in long-term survival in a select group of patients with this condition. The sequence of diagnostic tests and treatments for these patients are presented in a chronological order as a clinical pathway. This exposition is to provide a standardized management plan that will allow critical evaluation of the morbidity, mortality, outcomes and cost of this protracted plan of patient care. The discipline that the clinical pathway process demands may facilitate the continued optimization of this treatment strategy. PMID- 9349310 TI - Attitudes of Italian general practitioners in the treatment of cancer pain. The Committee of the Associazione Italiana di Oncologia Medica (AIOM). AB - The attitude of Italian general practitioners in prescribing practices for patients with cancer pain was assessed by means of a questionnaire. The results indicated that among most of the doctors who completed the questionnaire the basic principles of pain treatment in cancer patients are largely understood. Oral morphine emerged as the most commonly used opioid (60%) and controlled release morphine as the preferred preparation. Non-steroidal anti-inflammatory drugs were the most commonly used minor analgesics. Fear of side effects and restrictive prescribing regulations emerged as the most important barrier against adequate pain management. The survey emphasised the need for continued efforts in implementing specific educational programming for improvement in cancer pain management. PMID- 9349311 TI - Comparing two modalities of management of women with cytologic evidence of squamous or glandular atypia: early repeat cytology or colposcopy. AB - Early repeat cytology is recommended in most screening programs for cervical cancer in subjects with squamous or glandular abnormalities not amounting to neoplasia (atypical squamous cells of undetermined significance, ASCUS), but immediate colposcopy is also recommended in some countries, especially those where there is easy access to colposcopic facilities. We evaluated the cost effectiveness of the two procedures in a prospective study of women with cytologic ASCUS, invited to cytocolposcopic assessment after 6 months. Colposcopy directed biopsy was assumed as the gold standard, and the accuracy of colposcopy at 6 months was assumed to be equal to that of immediate colposcopy. Out of 874 compliers, punch biopsy was performed in 303 cases (34.7%), and 19 CIN2+ lesions were detected (CIN2 = 12, CIN3 = 6, microinvasive carcinoma = 1). Detecting 13 CIN2+ lesions at colposcopy required 874 colposcopies and 303 directed biopsies: the cost per CIN2+ lesion detected with the procedure was 2,749 US$. Detecting 15 CIN2+ lesions at repeat cytology required 874 cytologic examinations, 137 colposcopies, 64 directed biopsies, and 6 diagnostic large-loop resections, the latter being performed in subjects with high-grade squamous intraepithelial lesion and less severe lesions at punch biopsy: the cost per CIN2+ lesion detected with the procedure was 1,961 US$. The policy of repeat smear was more cost-effective than immediate colposcopy. According to such results, the protocol of the Florence screening program has been modified since October 1996. PMID- 9349312 TI - Accelerated bifractionated radiation with concurrent cisplatin administration in locally advanced head and neck cancer: a feasibility study. AB - AIMS AND BACKGROUND: To test the feasibility of accelerated interrupted twice daily radiation and concurrent cisplatin administration in patients with locally advanced head and neck cancer. PATIENTS AND METHODS: Nineteen patients with locally advanced head and neck cancer were treated with accelerated bifractionated radiation with concurrent administration of cisplatin. There were 18 men and 1 female with a median age of 60 years (range, 17-71) and median performance status of 90 (range, 80-100). Sixteen patients (85%) presented with stage IV disease. Primary site included the nasopharynx (n = 7), oropharynx (n = 5), hypopharynx (n = 1) and larynx (n = 6). Radiation consisted of two fractions of 1.6 Gy each daily, five times weekly to a total dose of 64 Gy. Cisplatin was administered at a dose of 100 mg/m2 on days 2 and 28 of the treatment period. RESULTS: Nine patients achieved a complete response (47%; 95% Cl, 25%-70%) and 5 a partial response (26%; 95% Cl, 7%-46%). Grade III-IV toxicity included leukopenia (16%), mucositis (26%), dry mouth (5%), weight loss (16%) and infection (5%). After a median follow-up of 27.11 months (range, 1-33 +), 9 patients have died. Median time to progression was 11 months (range, 1-32 +) and median survival 25 months (range, 1-32 +). CONCLUSIONS: Accelerated twice-daily radiation with concurrent cisplatin is effective in locally advanced head and neck cancer and can be safely given with manageable toxicity. PMID- 9349313 TI - Adenocarcinoma of the body and tail of the pancreas: is there room for adjuvant radiotherapy? AB - AIMS AND BACKGROUND: Adenocarcinoma of the body and tail of the pancreas is a rare malignancy with a poor prognosis. Few long-term survivors have been reported in the literature. The role of adjuvant treatment after curative resection has not yet been assessed. This retrospective study aims to describe the patterns of failure and the survival of 10 patients treated with resection and adjuvant radiotherapy. MATERIALS AND METHODS: From 1982 to June 1994, 10 patients with adenocarcinoma of the body and tail of the pancreas received adjuvant radiotherapy in our department. There were 4 females and 6 males, with a median age of 63 years (range, 45-77). The pT distribution was 2 pT1, 4 pT2, 4 pT3 and for pN it was 7 pN0 and 3 pN1. Four patients had stage I, 3 stage II and 3 stage III disease. All the patients underwent a resection: distal pancreatectomy in 7, partial resection of the body in 1, and total pancreatectomy in 2. Gross residual disease was present in 2 cases. Three patients received intraoperative radiotherapy up to a dose of 12-15 Gy. Postoperative radiotherapy was given in 9 patients with a dose ranging from 40 to 50 Gy (median, 45). One patient who received intraoperative radiotherapy had no postoperative radiotherapy. In 4 patients, chemotherapy with 5-fluorouracil was given during the first week of irradiation. RESULTS: Six patients experienced a local-regional relapse and 3 developed metastases. The median survival was 21 months. The 5-year overall survival was 15%. Eight patients died of progressive disease. One patient who presented with stage I disease was alive and free of disease at 24 months from diagnosis and, interestingly, one with stage III disease was alive at 111 months. No severe treatment-related complications were observed. CONCLUSIONS: As in carcinoma of the head of the pancreas, adjuvant radiotherapy should be considered as an adjuvant treatment of resected adenocarcinoma of the body and tail of the pancreas. Further evaluation is necessary to assess the role of intraoperative radiotherapy. PMID- 9349314 TI - Does concomitant chemoradiotherapy influence cosmetic outcome in conservative treatment of breast cancer? AB - PURPOSE: To evaluate retrospectively factors influencing the cosmetic outcome after conservative treatment for breast cancer. MATERIAL AND METHODS: From 1988 until 1992, 164 patients were treated with conservative surgery (quadrantectomy) and radiotherapy with 60Co (50 Gy on the whole breast) plus 10 Gy on the surgical bed (300 kV photons) for T1-T2 breast cancers; 46 patients (28%) received concomitant adjuvant chemotherapy (CMF schedule). Cosmesis evaluation was carried out after 24 to 108 months (median, 38 months). A logistic regression analysis was performed to identify independent variables influencing the aesthetic outcome. P values of 0.05 or less were considered significant. RESULTS: Univariate analysis showed that T2 versus T1 (P = 0.0102), lower quadrants site (P = 0.0002) and concomitant adjuvant chemotherapy (P = 0.0009) produced a worse aesthetic outcome. Multivariate analysis confirmed the same factors: tumor size (P = 0.0020), tumor site (P = 0.0150) and concomitant chemotherapy (P = 0.0024). CONCLUSIONS: The significant negative influence on the cosmetic outcome of concomitant adjuvant chemotherapy implies questions about the timing of radiotherapy and chemotherapy in breast cancer conservative treatment. PMID- 9349316 TI - Diagnosis of chromophobe renal cell carcinoma by chromosomal analysis. AB - Chromophobe renal cell carcinoma may pose a differential diagnostic problem by routine histologic examination because it may be misdiagnosed as another type of renal cancer with a totally different clinical behavior. A low DNA content as well as hypodiploidy seem to be associated with this renal tumor subtype. We report a case in which the cytogenetic report was of great value for a correct histologic diagnosis. PMID- 9349315 TI - The influence of blood glucose levels on [18F]fluorodeoxyglucose (FDG) uptake in cancer: a PET study in liver metastases from colorectal carcinomas. AB - AIMS AND BACKGROUND: To study the influence of blood glucose levels on the clinical reliability of positron emission tomography (PET) with [18F]-2-fluoro-2 deoxy-D-glucose (FDG) in the detection of liver metastases from colorectal carcinomas and in the analysis of tumor uptake of FDG. METHODS: After having given their informed consent, 8 patients with 20 liver metastases (mean size, 31.5 mm; range, 10-75 mm) detected by means of CT were submitted to a first FDG PET examination under fasting conditions and, 2 days later, to a second FDG-PET examination performed after i.v. infusion of a glucose solution (4 mg/kg/min for 2 hrs). The results of the two studies were compared in each patient, considering both the localization of the metastases and the FDG uptake in the lesions. A non kinetic method was used, calculating the Standardized Uptake Value (SUV). RESULTS: All 20 metastases were clearly visible on FDG-PET under fasting conditions. Moreover, in 2 patients FDG-PET detected a number of unknown liver metastases. The blood glucose levels after glucose infusion were significantly higher than the levels under fasting conditions, 158 +/- 13.8 mg/100 ml (mean +/- sd) and 92.4 +/- 10.2, respectively (P < 0.001), and the quality of the FDG-PET images showed a marked deterioration. FDG-PET was unable to detect 6 of the 20 lesions and another 10 lesions were localized less clearly. Moreover, 80% of the unknown liver metastases were not detected after glucose loading. The SUVs of metastases decreased from 9.4 +/- 5.7 (mean +/- sd) under fasting conditions to 4.3 +/- 8.3 after glucose loading (P < 0.001). CONCLUSIONS: FDG-PET studies may be particularly unreliable under conditions of high levels of blood glucose. Therefore, patients entering FDG-PET studies should fast, and blood glucose concentration needs to be taken into account when evaluating FDG uptakes in follow-up studies. PMID- 9349317 TI - Down-modulation of P210bcr/abl induces apoptosis/differentiation in K562 leukemic blast cells. AB - AIMS AND BACKGROUND: K562 cells are growth factor independent and neither function as stem cells nor differentiate into functional end cells. They are blast cells. There is evidence that the constitutively expressed bcr-abl tyrosine kinase might be responsible for the maintenance of the blast state of CML cells. We have studied the effect of two tyrosine kinase inhibitors, quercetin and genistein, on K562 cells. METHODS: K562 cells were treated with quercetin/genistein for a period of 72 hrs and then subjected to staining for apoptosis and erythroid differentiation and Western blotting with c-abl and phosphotyrosine monoclonal antibodies. RESULTS: The IC50 value was found to be 9.2 micrograms/ml for quercetin and 11.8 micrograms/ml for genistein. Quercetin treated cells did not show any differentiation but showed 68% apoptosis as compared to 7% in control. Genistein-treated cells showed 16% apoptosis and 15% erythroid differentiation. Quercetin reduced the level of p210 by 74% and its phosphotyrosine content by 67.6%. Genistein reduced p210 by 77.8% and its phosphotyrosine content by 16%. CONCLUSION: Both quercetin and genistein are able to down-modulate the tyrosine kinase activity of p210 as well as bring about a decrease in the content of the protein with different effects: quercetin induced apoptosis while genistein brought about both differentiation and apoptosis. PMID- 9349319 TI - Bilateral synchronous testicular involvement in multiple myeloma. Case report and review of the literature. AB - The authors describe a case of synchronous bilateral involvement of the testes in a 70-year-old patient seven years after the onset of an IgG k IIIA multiple myeloma. Ultrasonographic and postoperative immunohistochemical studies were performed. A complete review of the literature shows the rarity of testicular plasmacytoma. The present one is the second reported case of syncronous involvement of the testes. PMID- 9349318 TI - Immunophenotypes and cytotoxic functions of lymphocytes in patients with hepatocellular carcinoma. AB - AIMS AND BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common cancers in Asia. Immunological mechanisms are thought to play an important role in the control of tumor progression. The immune responses in HCC patients are poorly understood. In the present study, the proliferation and cytotoxic functions of lymphocytes from tumor tissues and peripheral blood of HCC patients were analysed. Simultaneously, the microcultures were phenotyped in order to determine the involvement of different lymphocyte subsets in mediating the cytotoxic function. METHODS: The frequencies of proliferating and cytotoxic lymphocytes from three tumor tissues and peripheral blood from ten HCC patients and nine healthy individuals were assessed by limiting dilution microculture analysis. These microcultures were phenotyped by single and dual color flow cytometry using monoclonal antibodies specific for CD4, CD8, CD56 and HLA-DR markers. RESULTS: The precursor frequencies of both proliferating and cytotoxic lymphocytes were found to be comparable in the peripheral blood of HCC patients and healthy individuals. Compared to peripheral blood, a marked reduction in the precursor frequencies of proliferating and cytotoxic lymphocytes was observed in the tumor tissues of HCC patients. In the tumor tissues, a significantly higher frequency of cytotoxic T cells compared to natural killer cells was observed. Dual color flow cytometric analysis revealed increased percentages of CD8+ HLA DR+ lymphocytes compared to CD4+ HLA-DR+ cells in the tumor tissues. CONCLUSIONS: Our results suggest that depressed immune responses at the tumor site might be responsible for the escape of tumor cells from the immune surveillance of the host. PMID- 9349320 TI - Angiotropic (intravascular) large cell lymphoma: case report and short discussion of the literature. AB - We report a case of angiotropic (intravascular) large B-cell lymphoma in an 84 year-old woman who underwent diagnostic procedures for progressive, painful induration of the legs. Physical examination and imaging studies revealed only widespread telangiectasias and significant panniculities-like lymphedema of the legs, with no masses or lymphadenopathies. The patient achieved a complete clinical remission after the first three cycles of polychemotherapy. Although angiotropic lymphoma is a rare entity with polymorphic clinical presentations, its early diagnosis appears very important since it may be curable with appropriate chemotherapy regimens. PMID- 9349321 TI - Carcinoma of the prostate: brain stem metastasis as the only site of spread. AB - AIMS AND BACKGROUND: Metastasis to the brain from prostate carcinoma is a rare event; it is reported in less than 4% of postmortem examinations. The prevalence of cases detected antemortem is even smaller, and the prevalence of brain stem metastasis as the only site of metastasis has been reported in only two other cases. METHOD: The authors present a third such case. RESULTS: A 55-year-old man, treated for an adenocarcinoma of the prostate (prostatectomy and radiotherapy), started to complain of facial expression disturbances and headaches 2 years later. Physical examination showed a left VII cranial nerve palsy. MRI showed an enhancing mass in the pons. Total body CT was negative. The patient was treated with a course of whole-brain and brain stem radiotherapy following stereotactic biopsy. Four months after radiotherapy, the neurological symptoms had disappeared and the patient died of a myocardial infarct. The systemic disease was still clinically silent. CONCLUSION: Our case involved only brain stem metastasis, probably implicating Batson's direct route of the paravertebral venous pathway. PMID- 9349322 TI - Splenomegaly as the first manifestation of thyroid cancer metastases. AB - We report the case of a 65-year-old man who developed a symptomatic splenomegaly due to spleen metastasis from thyroid follicular carcinoma. In 1982, at the age of 53, the patient had undergone a thyroid lobectomy for a cold node, followed one year later by a second intervention for a microfollicular adenoma. He was subsequently administered thyroid suppressive therapy with no further follow-up. The diagnosis of spleen metastases from thyroid cancer was first suspected on the basis of history, high serum thyroglobulin (Tg) levels, and the presence of pulmonary 99Tc uptake. The patient underwent a splenectomy, during which vast infiltration involving the diaphragm, spleen, stomach, colon and pancreas, was found. Histological and immunohistochemical results showed that the spleen and diaphragm metastases derived from thyroid follicular carcinoma. Radioiodine uptake by the pulmonary metastases confirmed the thyroid source. Retrospective re evaluation of the thyroid tissue removed in 1983 revealed a histological pattern consistent with follicular carcinoma, which could not be unequivocally attributed to the widely or minimally invasive form. To our knowledge this is the first report of splenomegaly as the first manifestation of thyroid cancer metastases. In this paper cases of splenomegaly due to metastatic spread are reviewed and the management of the present case is discussed. PMID- 9349323 TI - Thyroid metastases from a ductal carcinoma of the breast. A case report. AB - A case of bilateral thyroid metastases from ductal carcinoma of the breast is presented with emphasis on some unusual clinical findings: presentation after a long disease-free interval; clinical signs mimicking an acute thyroiditis; cystic structure of the left-sided metastatic nodule. Fine needle aspiration cytology from both nodules showed highly atypical tumor cells, such as to warrant a differential diagnosis between metastatic breast cancer on the one hand and anaplastic and medullary carcinoma of the thyroid on the other. Immunophenotypic study of the neoplasm on a cell block preparation of the aspirated material showed negativity for both thyroglobulin and calcitonin; instead, the tumor cells were strongly stained with antibodies against the "breast-related" markers alpha lactalbumin and gross cystic disease fluid protein-15. Therefore, immunochemistry allowed us to establish a definite diagnosis of metastatic breast disease of the thyroid, thereby avoiding surgical procedures. PMID- 9349324 TI - An appraisal of narrowband (TL-01) UVB phototherapy. British Photodermatology Group Workshop Report (April 1996). PMID- 9349325 TI - Antigen-independent expansion of T cells from psoriatic skin lesions: phenotypic characterization and antigen reactivity. AB - The pathogenesis of psoriasis appears to depend on T cells, which have been proposed to mediate the disease through an autoimmune process. To test this hypothesis we have propagated four T-cell lines from biopsies of psoriatic skin lesions by antigen-independent methods. Flow cytometric immunophenotyping showed the lines to be composed mainly of CD4-positive, alpha beta T-cell receptor (TCR) positive cells, which secreted a cytokine profile suggestive of predominant T helper type 1 (Th1) status. Analysis of TCR variable region (V beta) usage revealed two- to eight-fold increases in the expression of certain V beta species in lesional lines as compared with autologous peripheral blood mononuclear cells (PBMC), with the increased V beta species being expressed on more than 5% of cells in two of the lines. Lines were also used to test for responses to a range of epidermal antigen preparations in the presence of irradiated autologous PBMC as antigen-presenting cells. The lines failed--to proliferate in response to psoriatic lesional stratum corneum extracts, dispase-separated normal human epidermal extracts, and an epidermal keratin preparation before and after trypsinization, in spite of good proliferative responses to anti-CD3 which indicated that the lines were not anergic. In addition, the lines and PBMC from normal volunteers and the patients with psoriasis gave little or no response to recombinant streptococcal M protein. Thus, in spite of accumulating evidence for selective expansion of certain V beta-expressing T cells in psoriatic lesions, epidermal autoantigens have not been identified by using a bioassay which depended largely on the proliferation of lesional CD4-positive cells. The role of streptococcal M protein, which bears some homology with epidermal keratin is also open to question, at least in chronic plaque psoriasis. Further work is therefore required to obtain direct evidence that autoimmune processes are important in the pathogenesis of chronic plaque psoriasis. PMID- 9349326 TI - Mapping of monilethrix to the type II keratin gene cluster at chromosome 12q13 in three new families, including one with variable expressivity. AB - Monilethrix is an autosomal dominant disorder chiefly affecting hair. The degree of hair dystrophy is highly variable, as is the presence of additional features, such as follicular keratoses. In three British families of monilethrix, linkage has recently been reported to the type II keratin gene cluster at chromosome 12q13, and it has been suggested that the disease is due to a defect in the hard keratins of hair and nail. If monilethrix is a keratin disorder, we would predict that some pedigrees might map to the type I keratin gene cluster on 17q where hard keratin genes are also found. We have now studied clinically and by linkage analysis three new and unrelated pedigrees from England, Scotland and Spain, the first of which showed a variant phenotype. In this family the disease was expressed in four of 12 cases only as a follicular-keratosis of the neck, elbows and knees, and without clinical or historical evidence of hair anomalies; non penetrance in an obligate carrier was also observed. In all three families, we have established linkage to a series of microsatellite markers at the type II locus at 12q13 (Zmax = 6.34 at theta = 0.00 for D12S368) and have excluded linkage from the type I keratin gene cluster on 17q. It remains probable that monilethrix is a disorder of hard keratins, but at present there is no evidence that it is due to defects in type I keratins. PMID- 9349329 TI - Widespread microsatellite instability in sebaceous tumours of patients with the Muir-Torre syndrome. AB - Muir-Torre syndrome (MTS) is an autosomal dominant disorder characterized by the presence of at least one sebaceous gland tumour and a minimum of one visceral malignant tumour. Recently, microsatellite instability (MSI) has been detected in the tumours of patients with MTS and germline mutations of the hMSH2 and hMLH1 mismatch repair genes have been detected in some patients with this syndrome. To determine if the tumours of patients with MTS have widespread genomic instability and whether loss of heterozygosity (LOH) in the chromosomal regions containing hMSH2 and hMLH1 is detectable, MSI and LOH were examined at 10 dinucleotide repeats on chromosomes 2p, 3p, 5q, 9p, 17p and 18q. Data were obtained from six sebaceous gland tumours and two adenocarcinomas of the colon from three patients of two Muir-Torre families. MSI was detected at more than half of the loci tested in all sebaceous tumours examined. In addition, there was LOH at D2S119 in one sebaceoma and one sebaceous carcinoma from one patient. The colon carcinomas from two patients showed MSI at five of the 10 loci analysed. These results show that widespread MSI is a feature of tumours in patients with MTS. In addition, the finding of LOH at D2S119, a marker located in the vicinity of hMSH2, in sebaceous tumours of one patient indicates that this gene may have a pathogenetic role in this patient. PMID- 9349327 TI - Clinical features of photodamaged human skin are associated with a reduction in collagen VII. AB - Chronically sun-exposed or photodamaged human skin is characterized by a number of clinical features, including wrinkles. However, little is known about the molecular mechanisms that underlie these features. We investigated the hypothesis that the mechanism of wrinkle formation may involve loss of anchoring fibrils, composed mainly of collagen VII, which are important in maintaining dermal epidermal junction integrity. Ten volunteers with moderate to severe photodamage of dorsal forearm skin were recruited to the study. Using immunohistochemistry, transmission electron microscopy and in situ hybridization, we compared collagen VII protein and mRNA content of photodamaged forearm skin with that of sun protected hip and upper inner arm skin from the same subjects. Numbers of anchoring fibrils per linear microns of basement membrane (mean +/- SEM) were significantly lower in photodamaged skin (1.79 +/- 0.10) as compared with sun protected hip (2.28 +/- 0.11) and upper inner arm skin (2.21 +/- 0.10) (P < 0.01), and similarly keratinocyte expression of collagen VII mRNA, quantitated as number of positively stained keratinocytes per high power field, was significantly reduced in photodamaged skin (6.3 +/- 2.5) as compared with sun protected hip (20.0 +/- 5.6) and upper inner arm skin (17.7 +/- 4.9) (P < 0.001). Semiquantitative assessment of immunohistochemical staining for collagen VII showed a non-significant reduction in photodamaged skin as compared with sun protected skin. We propose that reduced content of collagen VII in photodamaged skin contributes to wrinkle formation by weakening the bond between the dermis and epidermis. PMID- 9349328 TI - Molecular determination of dermatophyte fungi using the arbitrarily primed polymerase chain reaction. AB - Dermatophytes are keratinophilic fungi capable of causing dermatophytosis (commonly known as tinea or ringworm) in humans and animals. Previously, we reported the differentiation of the common dermatophytes Trichophyton rubrum, T. mentagrophytes and T. tonsurans using a random primer 5'-ACCCGACCTG-3' (OPAA11) in the arbitrarily primed polymerase chain reaction (AP-PCR). In the present study, by examining additional dermatophytes including eight Microsporum spp., 16 Trichophyton species/subspecies and Epidermophyton floccosum using both OPAA11 and a second random decamer 5'-GAGAGCCAAC-3' (OPD18) in AP-PCR, we show that except for T. rubrum and T. gourvilli, and three T. mentagrophytes varieties, most of the dermatophyte fungi investigated formed distinct DNA band patterns on gel electrophoresis. The amplification of specific DNA bands in AP-PCR appeared to be independent of culture variations shown by dermatophyte isolates. These results provide the basis for the rapid identification of dermatophytes at the genetic level, supplementing existing laboratory methods and improving the diagnosis of human dermatophytosis. PMID- 9349330 TI - Expression of cathepsin D and B in invasion and metastasis of squamous cell carcinoma. AB - To determine the involvement of proteinases with hydrolytic activity towards extracellular matrix and basement membrane, in invasion and metastasis of tumour cells, the expression of cathepsin D, an aspartic proteinase, and cathepsin B, a cysteine proteinase, was studied. Formalin-fixed paraffin-embedded specimens from 13 patients who had squamous cell carcinomas (SCC) with local recurrence, skin and/or lymph node metastasis were examined. Cathepsin D stained intensely as a granular pattern (mature enzyme) in tumour cells of 69% of primary lesions and all the secondary lesions of the patients with SCC. Cathepsin B stained more intensely in SCC cells of all of the primary and secondary lesions than in normal epidermis; staining patterns were almost diffuse (procathepsin B). Granular and diffuse patterns (mature enzyme of cathepsin D and procathepsin B, respectively) appeared in the outer and inner parts of tumour islands, respectively. The presence of the active mature form of cathepsin D and procathepsin B in metastatic skin lesions of SCC was confirmed by Western blotting analysis. The presence and localization of the active mature form of cathepsin D suggests that activated cathepsin D may be involved in the invasion and metastasis of SCC. PMID- 9349331 TI - Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy. AB - Objective comparison of different antipsoriatic therapies requires quantitative assessment of disease severity. However, clinical assessment with the widely used Psoriasis Area and Severity Index (PASI) introduces inaccuracy. An alternative is the quantitative analysis of different epidermal cell parameters using multiparameter flow cytometry. Our aim in the present study was to compare the clinical and flow cytometric approach to monitor disease activity and to evaluate antipsoriatic efficacy. Clinical scores for erythema, induration and scaling were assessed and biopsies for flow cytometric analysis were obtained from the psoriatic plaques of 89 patients before and after treatment with different therapeutic regimens consisting of vitamin D3 analogues and corticosteroids. In total, 219 epidermal cell suspensions were analysed using triple-labelling, with the simultaneous staining of markers for epidermal proliferation (DNA dye TO-PRO 3), differentiation (antikeratin 10), and inflammation (antivimentin). Correlation analysis was performed on 166 paired values obtained from 83 patients. A highly significant correlation was observed between erythema and the percentage of vimentin-positive cells, between scaling and the percentage of keratin 10 positive keratinocytes, and between induration and the number of basal keratinocytes in the S- and G2M phase, when all 166 biopsies were assessed. The correlation remained in the same range if the analysis was restricted to the 83 pretreatment biopsies. In contrast to the clinical scores, the flow cytometric analysis permitted a clear separation between the antiproliferative and anti inflammatory or keratinization-enhancing effects of antipsoriatic treatment. The vitamin D3 analogues proved to exert a mainly antiproliferative effect. The combination of calcipotriol and a topical corticosteroid improved all cell biological markers substantially, and clobetasol monotherapy had a powerful effect on these markers. In conclusion, multiparameter flow cytometry has been shown to be a sensitive tool to evaluate the growth inhibiting, anti-inflammatory and keratinization-enhancing effects of antipsoriatic therapies. PMID- 9349332 TI - Presence of somatostatin in normal human epidermis. AB - Somatostatin (SOM) is a ubiquitous peptide which is responsible for the inhibition of numerous biological functions. SOM is described as an antiproliferative molecule and an inhibitor of exocrine or endocrine secretion from a variety of tissues, including pancreas, gastrointestinal tract, central and peripheral nervous system. Mediation of SOM effects can be indirect or direct, respectively, through other molecules or receptors on target cells. We have searched for the presence of SOM in the epidermis using immunofluorescence, confocal laser scanning microscopy, radioimmunoassay, and chromatography. Immunofluorescence and confocal laser scanning microscopy studies were performed using rabbit antiserum anti-SOM and mouse monoclonal antibody directed to CD1a Langerhans cell (LC) marker disclosed with fluorescein or tetramethylrhodamine isothiocyanate conjugates. SOM was extracted from whole skin or epidermal cell suspension or LC-enriched suspensions and analysed by radioimmunoassay. We used an antiserum which was reactive for the 6-11 portion of native SOM. Chromatographic columns were performed on extracts from whole skin. The epidermis was SOM immunoreactive. LC were immunoreactive for SOM and the staining was membranous. SOM was extracted from the whole skin at about 0.13 +/- 0.02 fmol/mg of tissue (mean +/- SEM). The SOM concentration in epidermal cell suspensions was 1.5 +/- 0.9 fmol/10(6) cells. Data obtained with LC-enriched suspensions showed large variations between donors. Extracts from skin showed one peak with an elution profile like that of 14 amino acid SOM. This study demonstrates that 14 amino acid SOM is expressed in normal human epidermis. PMID- 9349333 TI - Serum S-100 protein: a potentially useful prognostic marker in cutaneous melanoma. AB - S-100, an acidic calcium-binding protein, is present within cells of neuroendocrine origin. Its value in the immunohistochemical diagnosis of tumours of melanocytic origin is well established. More recently, a potential role has been proposed for the serum concentration of this protein as a marker of metastatic melanoma disease activity. In the present study, the concentration of serum S-100 protein was measured in 97 patients with histologically proven malignant melanoma who were attending a dermatology and/or oncology department for the follow-up of their disease. Serum S-100 was also measured in 48 control subjects without malignant melanoma. The clinical stage of the patients was classified according to the criteria of the American Joint Committee on Cancer into stages I-IV. The median (range) serum S-100 protein concentration was significantly higher in stage I (0.11 (0.1-0.21) microgram/L, P < 0.001), stage II (0.11 (0.05-0.22) microgram/L, P < 0.001), stage III (0.24 (0.07-0.41) microgram/L, P < 0.0001) and stage IV (0.39 (0.06-15.0) microgram/L, P < 0.0001) compared with the control group (0.1 (0.05-0.15) microgram/L). At a threshold value of 0.2 microgram/L, the sensitivity and specificity for detection of advanced disease were 82% and 91%, respectively. Thus serum S-100 protein may be a valuable prognostic marker for malignant melanoma and for monitoring therapy. Serum S-100 protein concentration was also compared with the Breslow thickness of the tumours. There was a significant correlation between these variables (n = 72, rs = 0.32, P < 0.01). Combining a serum S-100 threshold value of > 0.22 microgram/L and a Breslow thickness of > 4 mm improved the sensitivity and specificity for the presence of secondary spread to 91% and 95%, respectively. Therefore, a combination of both baseline serum S-100 protein and Breslow thickness may provide a better indication of the prognosis at diagnosis. PMID- 9349334 TI - The effects of topical doxepin on responses to histamine, substance P and prostaglandin E2 in human skin. AB - The tricyclic antidepressant, doxepin, is known to have H1 and H2 antihistaminic effects. Recently, 5% doxepin cream has been marketed in the U.S.A. for treatment of eczematous dermatoses. We investigated the effects of topical doxepin treatment on histamine-, substance P- and prostaglandin E2- (PGE2) induced responses in the skin of normal and atopic subjects. We compared the effects of topical doxepin with those of the oral antihistamine terfenadine. The weal volume and flare area responses to histamine were significantly reduced by treatment with topical doxepin or oral terfenadine in both normal and atopic subjects (P < 0.05). The mean +/- SEM percentage reduction in flare area for 10 micrograms/site of histamine in non-atopics and atopics was 48 +/- 8% and 60 +/- 17% with terfenadine, and 54 +/- 12% and 81 +/- 4% with topical doxepin, respectively. The mean percentage reduction in weal volume for the same dose of histamine in non atopics and atopics was 70 +/- 9% and 63 +/- 16% with terfenadine, and 96 +/- 2% and 89 +/- 6% with topical doxepin, respectively. The flare but not the weal response to substance P was inhibited by both treatments in all subjects (P < 0.05). The mean +/- SEM percentage reduction in flare area for 200 pmol/site of substance P in non-atopics and atopics was 53 +/- 10% and 73 +/- 4% with terfenadine, and 74 +/- 7% and 75 +/- 4% with topical doxepin, respectively. The cutaneous responses to PGE2 were not affected by either drug. The inhibitory effects of doxepin were as great as those of terfenadine, and doxepin had a significantly greater effect than terfenadine in inhibiting the weal response to histamine and flare response to substance P in normal volunteers (P < 0.05). There was no significant difference between atopics and non-atopics in the percentage reduction of cutaneous responses by oral terfenadine or topical doxepin. Marked sedation occurred in three of the first 10 subjects treated with topical doxepin, necessitating a reduction in dosage for the remaining six subjects. In summary, topical doxepin was as effective as, and sometimes more effective than, a standard dose of oral terfenadine in the inhibition of histamine-induced and axon-reflex-mediated cutaneous responses. The marked sedative effect may limit its clinical use in some patients. PMID- 9349335 TI - A prospective longitudinal study of the outdoor behaviour and symptoms of photosensitive patients. AB - The purpose of this study was to determine the measures that photosensitive patients use to control their sun exposure. Each week from March until September 1995, 30 patients with polymorphic light eruption (PLE) and 17 patients with chronic actinic dermatitis (CAD) returned a set of reply paid postcards on which they recorded information about their outdoor behaviour and symptoms. The principal differences between the two groups were that CAD patients had a much greater incidence of symptoms despite making more use of protective measures such as covering arms, wearing hats and applying sunscreen, than patients with PLE. And that as summer approached the PLE patients spent more time outdoors, whereas there was less seasonal variation in this respect among CAD patients. Tentative conclusions drawn from mathematical modelling indicated that the incidence of rash on a particular day was influenced by ambient ultraviolet radiation and length of time spent outdoors. There were indications that wearing a hat and keeping the arms covered offered some protection, whereas use of sunscreen may actually increase the likelihood of symptoms. PMID- 9349336 TI - Application of machine vision to assess involved surface in patients with psoriasis. AB - An objective method is still lacking in the quantitative assessment of psoriasis severity. The purpose of this study was to examine whether computer image analysis (CIA) by a new colour segmentation method can be used as an objective method of estimating involved surface area in patients with psoriasis. Involved surface area in psoriasis was assessed from colour photographs covering the same areas as the psoriasis area and severity index (PASI) scores of 26 patients by human observers from different medical professional groups and by CIA. The reliability of CIA was tested with reference analysis by colouring manually the involved areas in the images with the aid of projected slides. The surface area estimates by the different methods were compared with each other and the resulting effects on the PASI score were assessed. The agreement between the reference analysis and the image analysis was high, especially on trunk areas, but the cylindrical shape of the limbs resulted in some difficulties in assessing involved skin area. The human eye estimates differed from the image analysis ones in almost one-third of the cases and mostly when psoriasis affected under 30% of the skin surface area. Both over- and underestimates emerged, but overestimates were more common. Error estimates had a significant effect on the PASI score. The CIA method seems to be reliable and practicable in estimating the actual surface of psoriasis. The method has the disadvantage of being time-consuming (photographing, processing of pictures) and technically demanding. Further development of this method should make it faster in the future. PMID- 9349337 TI - Dermatoglyphics in Darier's disease. AB - Palmar and fingerprints were taken from 70 patients with Darier's disease and 409 normal controls. The dermatoglyphic characteristics of each group were determined and comparisons were made between them. Dermatoglyphic abnormalities were found; some confirmed previous findings and some were novel, but they are too small in degree and frequency to be of use diagnostically. PMID- 9349338 TI - The Q-switched neodymium:YAG laser and tattoos: a microscopic analysis of laser tattoo interactions. AB - The Nd:YAG laser effectively removes or lightens amateur and professional tattoos. The biomechanics of the removal of tattoo particles at the cellular level are incompletely understood. We examined biopsies obtained from 35 amateur and professional tattoos (including coloured tattoos), treated on three or more occasions with the Nd:YAG laser. Biopsies taken immediately after laser treatment showed vacuolation with complete clearance of tattoo particles in the most superficial layers of the dermis, as assessed by light and electron microscopy. We propose that the 'disappearance' of the tattoo particle arises from the formation of atomic species and gaseous products, which are rapidly dissolved in the extracellular fluid. Residual fragmented particles that are commonly found in the mid- and lower dermis are rephagocytosed. The interaction between the Nd:YAG laser and black tattoo particles at 1064 nm, and red tattoo particles at 532 nm, appears to be specific, as there was little evidence of thermal damage to adjacent cells or stromal collagen. PMID- 9349339 TI - The prevalence of seborrhoeic keratoses in an Australian population: does exposure to sunlight play a part in their frequency? AB - Although seborrhoeic keratoses (SKs) appear to be very common, there are very few studies reporting details of age-specific prevalence, distribution or possible cause. We report details on the frequency, nature and distribution of SKs in 100 Australian adults in the age groups 15-25, 26-50, 51-75 and those aged more than 75 years. There was an increase in prevalence of SKs from 12% of 15-25 year olds to 100% of those aged more than 50 years. The median number of lesions in those with them also increased with age from six per person in 15-25 year olds to 69 per person in those aged more than 75 years. There was no difference in prevalence or numbers of lesions/person between males and females. SKs on exposed areas were more often flat and more than 3 mm in diameter than those on the non exposed areas. There was a higher prevalence of SKs on the exposed areas than non exposed areas when taking into account the surface area. The data in this study demonstrate an increased frequency of SKs compared with those reported from the United Kingdom recently and from Australasia in the past, a phenomenon paralleling the changing frequency of skin cancer in these populations. This fact, plus the finding that SKs were more common as a function of skin surface area on the exposed areas of the body, suggests that sunlight may play a part in their development in those people who are predisposed to develop them. PMID- 9349340 TI - Skin testing of the pruritogenic activity of histamine and cytokines (interleukin 2 and tumour necrosis factor-alpha) at the dermal-epidermal junction. AB - Cytokines have been proposed as histamine-independent itch mediators. To investigate this hypothesis, single doses of interleukin-2 (IL-2, 10 MU/mL) and tumour necrosis factor alpha (TNF-alpha, 10 micrograms/mL) were delivered to the epidermis of 10 healthy volunteers with a controlled skin-prick model; 1% histamine and solvent controls were included in a double-blind, randomized crossover design. Itch ratings (computerized visual analogue scale) were obtained every 20 s for 15 min and cutaneous reactions (weal, flare and temperature) were measured. Reactions were also recorded after 2, 24 and 48 h. The mean itch ratings were: histamine 35.5, IL-2 3.3 (both P < 0.01 compared with control), TNF alpha 1.6 and solvent control 1.75 (not significant). Weal and flare occurred only with histamine. In two volunteers, an inflammatory papule with transient pruritus developed 12-18 h after applying IL-2. In conclusion, IL-2 showed a rapid, low pruritogenic effect, which may be followed by an inflammatory response. TNF-alpha induced no itching in this setting. Skin-prick testing with appropriate doses of potential pruritogens provides a safe and sensitive model for further chemoreceptor studies. PMID- 9349341 TI - The value of genotypic analysis in the assessment of cutaneous plasmacytomas. AB - We report a 79-year-old woman with primary cutaneous plasmacytoma in whom polymerase chain reaction (PCR) was used to demonstrate the monoclonality of the tumour. Four years after presentation, further skin lesions occurred and PCR again showed evidence of monoclonality despite the histology being non-specific. The reported frequency of multiple primary cutaneous plasmacytoma is increasing, and the mortality rate of patients with multiple lesions is three times that of those with a solitary lesion. PCR may be a useful technique for assessing such patients at presentation. PMID- 9349342 TI - Generalized telangiectasia as the major manifestation of angiotropic (intravascular) lymphoma. AB - We report a 60-year-old woman with B-cell angiotropic lymphoma who presented with generalized telangiectasia and indurated plaques. Cutaneous involvement in angiotropic lymphoma has a wide clinical spectrum. Telangiectasia on associated cutaneous lesions is expected, but generalized telangiectasia over normal skin was a striking feature of our patient. PMID- 9349343 TI - Epstein-Barr virus-associated lymphoproliferative eruption with progression to large granular lymphocytic leukaemia. AB - A 12-year-old Korean girl gave a 9-year history of recurrent necrotizing papules and vesicles on the face, scalp and extremities. Skin biopsy specimens showed an atypical lymphoreticular infiltrate with vasculitis in the dermis and subcutis. In situ hybridization demonstrated latent infection by Epstein-Barr virus (EBV) of the lymphoid cells in the dermis. The disease was diagnosed as an EBV associated lymphoproliferative skin eruption presenting as recurrent necrotic papulovesicles. The patient subsequently developed large granular lymphocytic leukaemia of natural killer cell origin. Our observations suggest that a patient with an EBV-associated lymphoproliferative skin eruption presenting with recurrent necrotic papulovesicles might progress to develop leukaemia as well as lymphoma. PMID- 9349344 TI - Chronic actinic dermatitis associated with human immunodeficiency virus infection. AB - Chronic actinic dermatitis is a photodistributed, eczematous dermatitis that preferentially affects elderly men and persists for months to years. Its occurrence in individuals infected with human immunodeficiency virus (HIV) has been described in five patients. We report four additional cases of this uncommon, chronic photodermatosis associated with HIV infection. In two of the patients, photosensitivity was a presenting disorder leading to the diagnosis of HIV infection. All patients were men of skin type VI with a mean age of 50 years, all had decreased minimal erythema doses to ultraviolet B, three of the four patients had decreased minimal erythema doses to ultraviolet A and all had CD4 cell counts of < 200 x 10(6)/L. PMID- 9349345 TI - Atypical Lyme borreliosis in an HIV-infected man. AB - We report the fourth case of Lyme borreliosis in a man infected with human immunodeficiency virus (HIV). The erythema chronicum migrans was persistent, overlapping with meningoradiculitis. Repeated immunofluorescence tests for Borrelia burgdorferi sensu lato remained negative in both sera and cerebrospinal fluid (CSF), the enzyme-linked immunosorbent assay was weakly positive in serum and CSF and a Western blot was positive. The skin infiltrate was composed mostly of T lymphocytes with a CD4/CD8 ratio of 0.5. The course of the disease was favourable after treatment with intravenous ceftriaxone. Further studies are necessary to evaluate whether HIV infection influences, as does syphilis, the course and response to treatment of Lyme borreliosis. Serological tests are insufficiently sensitive and the Western blot assay is necessary to confirm Lyme disease in HIV-positive patients. PMID- 9349346 TI - Herpes simplex virus type 2 infection in a 5-year-old boy presenting with recurrent chest wall vesicles and a possible history of herpes encephalitis. AB - A 5-year-old hyperkinetic but otherwise healthy child presented with recurrent irritable vesicles and erosions of the anterior chest wall; they have been apparent since the age of 15 months. Wound swab cultures yielded herpes simplex virus type-2 (HSV-2) and Western blot serology showed past exposure to both HSV-1 and HSV-2. Skin biopsy results further supported a herpes virus infection. Magnetic resonance imaging of the brain showed right temporal lobe atrophy. An evaluation showed no evidence of sexual abuse in the patient but a Western blot assay of the mother's serum for HSV-2 was positive, while the father's was negative. In view of the diagnosis of HSV-2 infection in such a young patient, the possible routes of transmission and the time of acquisition of infection were explored. We believe the most likely route of infection in this child was postnatal, through intimate contact with the mother. PMID- 9349347 TI - Non-lethal junctional epidermolysis bullosa in a dog. AB - We report a 4-year-old female mongrel dog with a history since birth of erosions and atrophic skin, with pigmentation and alopecia on the face, trunk and extremities, together with dystrophic nails. Light microscopy revealed subepidermal blisters with minimal inflammation and electron microscopy confirmed that the ultrastructural site of separation site was at the lamina lucida. Indirect immunofluorescence of the dog's skin detected the positive expression of laminin 5, BPAG2, integrin-alpha 6 and type VII collagen. These clinical, ultrastructural and immunohistochemical features suggested that the dog had a non lethal subtype of junctional epidermolysis bullosa (JEB). This is the first confirmed case of non-lethal JEB in a dog and presents a possible candidate for an animal model of gene therapy. Further study should provide important information of the phenotype, pathophysiology and prognosis of non-lethal JEB in humans. PMID- 9349348 TI - Massive cutaneous mucinosis associated with systemic lupus erythematosus. AB - We describe a 23-year-old Japanese man with systemic lupus erythematosus (SLE) who developed massive cutaneous mucinosis. He was diagnosed with SLE when he was 11 years old. Prednisolone therapy (30 mg/day) was initiated and reduced to 10 mg/day 3 months later; the SLE had been well-controlled with this dose of prednisolone for 12 years. However, infiltrated erythematous plaques developed on the middle-lateral area of his back at 17 years old and progressed to erythematous and elastic soft tumorous masses over 20 cm in diameter at 23 years old. Biopsies of the lesions on the nape revealed massive mucin deposition. Topical injection with hyaluronidase decreased the lesion. This cutaneous mucinosis can be distinguished clinically and histopathologically from papular and nodular mucinosis associated with SLE. The present case might be an unusual clinical variant of cutaneous mucinosis associated with SLE. PMID- 9349349 TI - Multiple glomus tumours, Coats' disease and basic fibroblast growth factor. AB - We report a patient with multiple glomus tumours and Coats' disease of the retina, an association not previously described. We discuss the possible pathogenesis of these conditions in view of the finding of elevated serum levels of basic fibroblast growth factor. PMID- 9349350 TI - Melanoma-associated paraneoplastic retinopathy: case report and review of the literature. AB - A 51-year-old white male suffering from metastatic malignant melanoma of the skin presented with lymph node metastases and paraneoplastic retinopathy 4 years after resection of the primary tumour. There were no cerebral metastases. Ocular symptoms consisting of night blindness and sensations of 'shimmering lights' persisted after total resection of the inguinal lymph node metastases and administration of dacarbazine and prednisone. Perimetry of both eyes was abnormal with concentric restriction. Electroretinography showed significantly reduced amplitudes in both eyes. Only 11 patients with melanoma-associated retinopathy (MAR) have been described. High titres of autoantibodies against whole retina extract were detected by enzyme-linked immunosorbent assay (ELISA) reactions. Indirect immunohistochemistry showed strong autoantibody activity against retinal bipolar cells. PMID- 9349351 TI - Fibroelastolytic papulosis of the neck: a report of 20 cases. AB - The clinical and histological features of the entities known as 'white fibrous papulosis of the neck' (WFPN) and 'acquired elastolysis of the papillary dermis simulating pseudoxanthoma elasticum' (PDE) are not clearly defined. This study was conducted to compare our experience of WFPN/PDE with those described in the literature. Twenty patients presented at our institution with papular eruptions involving the neck. The asymptomatic lesions, which ranged in colour from normal skin tones to yellowish, were isolated or coalescent. Microscopically, the papules showed elastolysis and fibrosis of the upper reticular and papillary dermis. A review of the literature shows similar characteristics in cases reported as WFPN and PDE. This study indicates that WFPN and PDE are variants of a single disorder that can be more precisely defined as 'fibroelastolytic papulosis of the neck' and which appears to be a manifestation of intrinsic skin ageing. PMID- 9349352 TI - Human herpesvirus 8 DNA sequences in angiosarcoma of the face. PMID- 9349353 TI - Comel-Netherton syndrome. A diagnostic challenge. PMID- 9349354 TI - Lymphocytoma cutis induced by cobalt. PMID- 9349355 TI - Nail changes as the only skin abnormality in myeloma-associated systemic amyloidosis. PMID- 9349356 TI - Acquired perforating dermatosis showing the Koebner phenomenon. PMID- 9349357 TI - Pseudoxanthoma elasticum: a study of urinary glycosaminoglycan levels in two cases. PMID- 9349358 TI - Bullous lichen sclerosus with chronic hepatitis C virus infection. PMID- 9349359 TI - Cutaneous vasculitis with partial C4 deficiency responsive to dapsone. PMID- 9349360 TI - Silica granuloma: another cause of tennis elbow. PMID- 9349361 TI - Cicatricial pemphigoid treated with intravenous immunoglobulin. PMID- 9349362 TI - Post-adolescent acne and marital break-up. PMID- 9349363 TI - Wells' syndrome responsive to PUVA therapy. PMID- 9349364 TI - Eczematous reactions to human immune globulin. PMID- 9349365 TI - Pavlovian conditioning in cold-exposure tests in Raynaud's phenomenon: pitfalls in diagnosis. PMID- 9349366 TI - Tinea nigra secondary to Exophiala werneckii responding to itraconazole. PMID- 9349368 TI - London medicine. PMID- 9349367 TI - Emergency care outside hospital. PMID- 9349369 TI - Management of thyroid tumours. AB - True tumours of the thyroid gland, both benign and malignant, are uncommon. Most present as solitary thyroid nodules and can be readily diagnosed by fine-needle aspiration and cytology. Surgical resection is the only appropriate treatment for the majority of thyroid tumours. PMID- 9349370 TI - Pathophysiology and management of portal hypertension. 2: Cirrhotic ascites. AB - The pathophysiology of the haemodynamic and renal abnormalities in cirrhosis remains ill-defined. The development of ascites has poor prognostic significance and management should follow a stepwise approach from salt restriction to diuretic therapy then large-volume paracentesis before more invasive techniques. PMID- 9349371 TI - Anaesthetic management of the severely injured patient: head injury. PMID- 9349372 TI - The diagnosis of pleural disease. PMID- 9349373 TI - Thoracoscopic surgery: past, present and future. PMID- 9349374 TI - Diagnostic imaging in pulmonary embolic disease. PMID- 9349375 TI - Ropivacaine. AB - Ropivacaine is a new aminoamide local anaesthetic drug. Its clinical profile is similar to that of bupivacaine but it causes less motor block and is less cardiotoxic. PMID- 9349376 TI - Resuscitation. II: Advanced cardiac life support. AB - Advanced cardiac life support is the definitive sequence of initial treatment for the victim of a cardiac arrest. Advanced cardiac life support forms a large part of cardiopulmonary resuscitation, which also includes basic life support and post resuscitation intensive care. PMID- 9349377 TI - Diagnosis of presenile dementia. AB - It may surprise some that presenile dementia has an incidence of 7.2 per 100,000 resulting in approximately 18 new cases per average district general hospital per year. The diagnosis and management of these patients is commonly divided among a number of different medical disciplines leading to varied clinical approaches to the problem. This article will outline an approach adopted at a cognitive function clinic and will emphasize the utility of a multidisciplinary team. PMID- 9349378 TI - Acceleration of leukaemia with granulocyte colony-stimulating factor in a patient with aleukaemic leukaemia. PMID- 9349379 TI - ATP and the role of IK.ATP during acute myocardial ischemia: controversies revive. PMID- 9349380 TI - Cardiomyopathies in disorders of oxidative metabolism. AB - Primary cardiomyopathy is an important cause of mortality in children and adults. Apart from inherited disorders of myocardial contractile and structural proteins, several defects of energy metabolism may cause cardiomyopathy. Most of the energy required for myocardial contraction is derived from aerobic metabolism. Faulty aerobic metabolism involving the heart may be due to defects of mitochondrial oxidative phosphorylation or to defects of fatty acid oxidation. Considerable advances have been made in the last 10 years in understanding the biochemical and molecular characteristics of mitochondrial disorders. Several point mutations or large-scale re-arrangements of mitochondrial DNA have been identified in patients with cardiomyopathy, either as part of complex multisystem syndromes or as the main clinical feature. Inborn errors of fatty acid oxidation are reported with increasing frequency as a cause of metabolic dysfunction, myopathy, cardiomyopathy, and sudden death in childhood. Advances in biochemical and molecular genetic techniques have considerably improved our understanding of the metabolic disorders causing cardiomyopathy, providing new tools for classification and diagnosis of candidate patients. The present review focuses on defects of mitochondrial oxidative metabolism associated with cardiomyopathy. PMID- 9349381 TI - Dealing with a modern-day epidemic. PMID- 9349382 TI - Monitoring fibrillar collagen turnover in hypertensive heart disease. PMID- 9349383 TI - Postextrasystolic left ventricular isovolumic pressure decay is not monoexponential. AB - OBJECTIVE: The relationship between the left ventricular (LV) relaxation time constant and early diastolic filling is not fully defined. This study provides additional evidence that LV isovolumic pressure fall in the normal intact heart in response to certain interventions is not adequately described by a model of monoexponential decay and that its relationship to filling is complex. METHODS AND RESULTS: To gain further insight into the relationship between LV relaxation and early rapid filling we measured LV isovolumic relaxation rate, peak early filling velocity (E), LV volumes, and transmitral pressures at baseline and in the first postextrasystolic beat after a short-coupled extrasystole in 9 anesthetized dogs. Postextrasystolic isovolumic relaxation rate was slowed as measured by 3 commonly used time constants, while E was increased 32%. LV contractility and peak pressure were also increased, while LV end-systolic volume was decreased. LV minimum pressure was deceased, while the early diastolic transmitral pressure gradient was increased. Although all relaxation time constants measured over the entire isovolumic relaxation phase indicated slowed relaxation, direct measurement of isovolumic relaxation time indicated no change in relaxation rate. Calculation of the time constants and direct measurement of isovolumic relaxation time during early isovolumic pressure decay indicated slowed postextrasystolic pressure decay rate compared with baseline, while calculation of time constants and direct measurement of isovolumic relaxation time during late isovolumic relaxation indicated augmented postextrasystolic pressure decay rate versus baseline. CONCLUSIONS: This non-exponential behavior of LV isovolumic pressure decay in postextrasystolic beats after short-coupled extrasystoles provides further evidence that the relationship that exists between ventricular relaxation and early filling is not simple. The results are interpreted in terms of current theoretical formulations that attribute control of myocardial relaxation to the interaction between inactivation-dependent and load-dependent mechanisms. PMID- 9349384 TI - Contribution of hypoxia to the development of cardiomyopathy in hamsters. AB - OBJECTIVE: It has been hypothesized that microvascular spasms cause cardiomyopathy. To elucidate the contribution of hypoxia to the development of cardiomyopathy, the newly-developed hypoxia tracer, Tc-99m nitroimidazole, was applied to detect myocardial hypoxia in a hamster model. METHODS: Tc-99m nitroimidazole (180 MBq) and I-125 iodoantipyrine (370 kBq) were injected into cardiomyopathic Syrian hamsters or control hamsters at age 10, 25, and 40 weeks (n = 6 in each group). The myocardial uptake of Tc-99m nitroimidazole was measured and dual tracer autoradiography was performed. RESULTS: Histologic study revealed that the cardiomyopathic hamsters at age 10, 25 and 40 weeks were in the myocytolytic stage, the fibrotic and healing stage, and the hypertrophy and dilatation stage, respectively. Tc-99m nitroimidazole uptake was significantly greater in the cardiomyopathic hamsters than in the controls at age 25 weeks (cardiomyopathic hamsters, 33.3 +/- 4.7% g dose/g; control, 25.2 +/- 3.1), whereas there were no significant differences between both strains at age 10 and 40 weeks. The quantified concentration of I-125 iodoantipyrine in the cardiomyopathic hamster at age 40 weeks was significantly lower than that in the controls. When the Tc-99m nitroimidazole uptake was normalized by I-125 iodoantipyrine concentrations, the cardiomyopathic hamsters at age 25 and 40 weeks showed significantly greater uptake than the controls. CONCLUSION: The myocardium in cardiomyopathic hamsters was hypoxic at the fibrotic and healing stage and may be hypoxic at the hypertrophy and dilatation stage. This may contribute to the development of cardiomyopathy. PMID- 9349385 TI - Effect of AT1 receptor blockade on cardiac collagen remodeling after myocardial infarction. AB - OBJECTIVE: Previous work has shown that cardiac fibrosis occurs after myocardial infarction (MI) in non-infarcted ventricular tissue and that this event is associated with abnormal cardiac function. Our aim was to investigate the effect of AT1 receptor blockade on cardiac collagen remodeling in post-MI rat heart remote from the infarct site by addressing collagen mRNA abundance, posttranslational hydroxylation of collagen monomers, and mature collagen deposition. Prolyl 4-hydroxylase (PH) mediates hydroxylation of procollagen alpha chains in the endoplasmic reticulum of cardiac fibroblasts and thus regulates the downstream formation and secretion of helical procollagen molecules. METHODS: The effects of losartan (15 mg/kg/day) on collagen deposition and mRNA abundance were monitored in viable left and right ventricles in sham-operated (control) and experimental groups in the presence or absence of losartan. Immunoreactive PH concentration in viable tissues as well as cardiac function in control and experimental groups was determined by ELISA. RESULTS: Immunohistochemical staining and 4-hydroxyproline assays confirmed that losartan treatment attenuates fibrosis in experimental hearts. Northern analysis revealed that losartan treatment of 1, 2, or 4 week experimental groups had no effect on collagen mRNA abundance compared to untreated post-MI rats. On the other hand, immunoreactive PH concentration was significantly decreased in the post-MI group treated with losartan. Determination of cardiac mass and cardiac function revealed that losartan treatment was associated with attenuated cardiac hypertrophy and improved left ventricular (LV) function in experimental animals. CONCLUSIONS: AT1 blockade is associated with a significant decrease in cardiac fibrosis in treated post-MI rats, and this trend is positively correlated to a significant decrease in immunoreactive PH compared to untreated experimental animals. The expression of cardiac PH may be regulated by angiotensin via AT1 receptor activation, and the suppression of PH with losartan treatment may be an important mechanism for modulation of collagen deposition in the post-MI rat heart. PMID- 9349386 TI - Targeting of dobutamine to ischemic myocardium without systemic effects by selective suction and pressure-regulated retroinfusion. AB - OBJECTIVE: To study the effects of low-dose dobutamine and/or glyceryl trinitrate in addition to selective suction and pressure-regulated retroinfusion with arterial blood on regional myocardial function of the ischemic myocardium and systemic hemodynamics. METHODS: Using a pig model of repeated brief (90 s) occlusions of the left anterior descending artery, selective suction and pressure regulated retroinfusion was carried out either with arterial blood alone (SSRalone) or with arterial blood and simultaneous application of low-dose dobutamine (0.1 microgram/kg/min (SSRDOB), glyceryl trinitrate (0.03 mg/kg/min) (SSRNIT) or the combination of both drugs (SSRDOB + NIT). Regional myocardial function of the ischemic and non-ischemic myocardium was determined by sonomicrometry (segment shortening). RESULTS: Segment shortening in the ischemic area after 90 s of ischemia was preserved at 57.5 +/- 9.2% with SSRalone but at 78.0 +/- 22.3% of baseline with SSRDOB (P < 0.05). The addition of glyceryl trinitrate did not improve regional myocardial function further. No effects of locally applied dobutamine were observed with regard to non-ischemic myocardium or heart rate. Cardiac output and mean arterial blood pressures tended to be further stabilized with SSRDOB. CONCLUSIONS: Local application of low-dose dobutamine together with arterial blood by selective suction and pressure regulated retroinfusion during brief myocardial ischemia resulted in improved regional myocardial function without undesired effects on non-ischemic myocardium or systemic hemodynamics. PMID- 9349388 TI - Interactive effect of the p53 gene and cigarette smoking on coronary artery disease. AB - OBJECTIVE: p53 is a tumour suppressor protein involved in the control of cell growth and has an established role in carcinogenesis, particularly in relation to smoking. It may also be related to arteriosclerosis by affecting smooth muscle cell proliferation, a feature of atherogenesis. METHODS: We explored a role for p53 in atherogenesis by assessing the association between two DNA polymorphisms of the p53 gene (MspI at intron 6 and HaeIII at intron 1) and angiographically documented coronary artery disease (CAD) in 654 Australian Caucasian patients. RESULTS: There was a significant interactive effect of the two polymorphisms and cigarette smoking on CAD in a logistic regression analysis (P = 0.0039) but no association between CAD and either individual p53 polymorphic marker. CAD occurrence was more frequent in non-smoking patients with rare alleles at both sites (85.0%) compared to those homozygous for common alleles at both sites (70.4%). However, this was not seen in smokers (85.7 vs 82.8%). In all 654 patients cigarette smoking remained a significant predictor of CAD irrespective of p53 genotypes (P = 0.0065). CONCLUSIONS: Our findings identify an interactive effect of both p53 polymorphisms and cigarette smoking on the occurrence of coronary artery disease in that non-smoking patients with rare alleles at both sites had increased incidence of CAD. They illustrate the relevance of genotype specific and environment-dependent enhanced cardiovascular risk and foreshadow a need for further studies to establish functional changes. PMID- 9349387 TI - Arterial expression of the plasminogen activator system early after cardiac transplantation. AB - OBJECTIVES: Recent studies suggest that alterations in tissue thrombolysis as well as the inward migration of cells may be specific events that contribute to coronary artery narrowing after cardiac transplantation. Plasminogen activators and inhibitors play a central role in governing not only tissue thrombolysis, but also vascular cell migration. The purpose of this study was to examine arterial wall expression of the plasminogen activation system in coronary arteries during graft vascular disease initiation and progression. METHODS: Using in situ hybridization and immunocytochemistry, the expression patterns of uPA and PAI-1 in coronary arteries from cardiac allografts were compared to those of young individuals without disease. RESULTS: Both PAI-1 and uPA were over-expressed early after transplantation and as late as 27 months post grafting. Over expression of these molecules preceded morphological evidence of graft vascular disease. Of special note was the adventitial expression of uPA and PAI-1 in microvessels and myofibroblasts. In contrast, the expression of uPA and PAI-1 in normal coronary arteries was confined to endothelial cells of the central lumen, as well as low levels of expression in intimal and medial smooth muscle cells. CONCLUSIONS: Despite morphologic similarities between normal and transplant coronary arteries, differences were noted in the vascular expression pattern of uPA and PAI-1. The exact role of these molecules in graft vascular disease requires further study; however, it is intriguing to consider that a local imbalance in the plasminogen system may contribute to arterial wall thrombosis and/or excessive cell migration and the genesis of complex vascular lesions. PMID- 9349389 TI - Electrophysiologic effects of acute myocardial ischemia: a theoretical study of altered cell excitability and action potential duration. AB - OBJECTIVE: To study the ionic mechanisms of electrophysiologic changes in cell excitability and action potential duration during the acute phase of myocardial ischemia. METHODS: Using an ionic-based theoretical model of the cardiac ventricular cell, the dynamic LRd model, we have simulated the three major component conditions of acute ischemia (elevated [K]o, acidosis and anoxia) at the level of individual ionic currents and ionic concentrations. The conditions were applied individually and in combination to identify ionic mechanisms responsible for reduced excitability at rest potentials, delayed recovery of excitability, and shortened action potential duration. RESULTS: Increased extracellular potassium ([K]o) had the major effect on cell excitability by depolarizing resting membrane potential (Vrest), causing reduction in sodium channel availability. Acidosis caused a [K]o-independent reduction in maximum upstroke velocity, (dVm/dt)max. A transition from sodium-current dominated to calcium-current dominated upstroke occurred, and calcium current alone was able to sustain the upstroke, but only after sodium channels were almost completely (97%) inactivated. Acidic conditions prevented the transition to calcium dominated upstroke by acidic reduction of both sodium and calcium currents. Anoxia, simulated by lowering [ATP]i and activating the APT-dependent potassium current, IK(ATP), was the only process that could decrease action potential duration by more than 50% and reproduce AP shape changes that are observed experimentally. Acidic or anoxic depression of the L-type calcium current could not reproduce the observed action potential shape changes and APD shortening. Delayed recovery of excitability, known as 'post-repolarization refractoriness', was determined by the voltage-dependent kinetics of sodium channel recovery; Vrest depolarization caused by elevated [K]o increased the time constant of (dVm/dt)max recovery from tau = 10.3 ms at [K]o = 4.5 mM to tau = 81.4 ms at [K]o = 12 mM, reflecting major slowing of sodium-channel recovery. Anoxia and acidosis had little affect on tau. CONCLUSIONS: The major conditions of acute ischemia, namely elevated [K]o, acidosis and anoxia, applied at the ionic channel level are sufficient to simulate the major electrical changes associated with ischemia. Depression of membrane excitability and delayed recovery of excitability in the single, unloaded cell are caused by elevated [K]o with additional excitability depression by acidosis. Major changes in action potential duration and shape can only be accounted for by anoxia-dependent opening of IK(ATP). PMID- 9349390 TI - Anoxia-induced activation of ATP-sensitive K+ channels in guinea pig ventricular cells and its modulation by glycolysis. AB - OBJECTIVE: Exposure to anoxia has been reported to activate ATP-sensitive potassium (K+(ATP)) channels in isolated ventricular myocytes. We aimed to investigate the mechanisms underlying the anoxia-induced activation of K+(ATP) channels. METHODS: Guinea pig ventricular myocytes were isolated using collagenase digestion. Action potentials and membrane currents were recorded in the whole-cell mode of patch clamp. Exposure to anoxia was performed in a semi closed airtight chamber, which prevented the diffusion of atmospheric oxygen into anoxic perfusate. RESULTS: Exposure to glucose-free anoxia shortened the action potential duration (APD) to less than 20% of control in 13 +/- 3 min. Subsequent reoxygenation rapidly and completely restored the APD. The time-independent large outward current which developed during anoxia was completely suppressed by reoxygenation or by the application of glibenclamide, a K+(ATP) channel blocker. The presence of extracellular glucose did not prevent APD shortening during anoxia, although it significantly decreased the rate of shortening. Reoxygenation induced restoration of the APD was inhibited after a long-lasting anoxia. In addition, repeated exposures to anoxia/reoxygenation progressively impaired the recovery of APD during reoxygenation. CONCLUSIONS: Activation of K+(ATP) channels occurs during anoxia. The primary source of ATP that regulates the channel activity seems to be oxidative phosphorylation. ATP derived from anaerobic glycolysis (attained by the increase of extracellular glucose) was observed to partially suppress the channel activity only when oxidative phosphorylation was severely impaired during anoxia. PMID- 9349391 TI - Effects of wortmannin on insulin- and ischemia-induced stimulation of GLUT4 translocation and FDG uptake in perfused rat hearts. AB - OBJECTIVE: Myocardial glucose transport is enhanced by hormonal and other stimuli such as ischemia and hypoxia which induce glucose transporter 4 (GLUT4) translocation. Whether insulin and ischemia share a common signaling mechanism is not yet known. This study investigated whether phosphatidylinositol 3-kinase (PI3K), a signaling intermediate of the insulin-responsible pathway, also participates in the ischemia-induced stimulation of glucose. METHODS: Isolated Langendorff-perfused Sprague-Dawley rat hearts were subjected to 100 nmol/l insulin or 15 min of no-flow ischemia with/without 1 mumol/l wortmannin, an inhibitor of PI3K. After perfusion, relative subcellular glucose transporter GLUT4 distribution was assessed by membrane fractionation and immunoblotting and compared to controls. Uptake kinetics of the glucose analog [18F]fluoro deoxyglucose (FDG) were also studied during perfusion of rat hearts. RESULTS: GLUT4 translocation to the plasma membrane (PM) was increased by insulin 1.8-fold and by ischemia 2.4-fold (P < 0.05). FDG uptake was increased by insulin 6.0-fold and by ischemia 6.2-fold (P < 0.05). Wortmannin 1 mumol/l inhibited insulin mediated translocation of GLUT4 and increase in FDG uptake completely. However, it did not show any effect on ischemia-stimulated GLUT4 translocation or on ischemia-induced increase in FDG utilization. A significant correlation was found between relative GLUT4 translocation and FDG uptake in hearts of the insulin series (r = 0.9, P < 0.05) and of the ischemia series (r = 0.8, P < 0.05). CONCLUSIONS: Our results demonstrate that wortmannin did not inhibit ischemia induced stimulation of myocardial glucose transport, supporting the hypothesis of different signaling pathways for ischemia and insulin. PMID- 9349393 TI - Acceleration of stiffness in underperfused diabetic rat hearts by glyburide, a KATP channel blocker, and its prevention by levcromakalim and insulin. AB - OBJECT: To clarify the role of the KATP channels in myocardial dysfunction during underperfusion with norepinephrine (NE) in the diabetic heart, particularly the heart treated with sulphonylurea derivatives. METHODS: Isolated 6-week streptozotocin-diabetic rat hearts with a balloon in the left ventricle (LV) were paced and perfused with normoxic Krebs-Henseleit solution. Agents were infused for 15-25 min before as well as during 60-min underperfusion (2 ml/min/g heart weight) with 10(-6) M NE. Regional myocardial flow distribution was measured using dye microspheres. The effects of ex vivo glyburide (10(-6) M, a sulphonylurea anti-diabetic drug and a specific KATP channel inhibitor) on contractile dysfunction and abnormal regional myocardial energy metabolism were examined during underperfusion with NE in the absence of presence of levcromakalim (10(-4) M, a selective K+ channel opener) and insulin (2 mU/min/g heart weight). RESULTS: The flow rate was greater in the LV subendocardium than the subepicardium during normal perfusion, and smaller at 60-min underperfusion with NE. The LV diastolic tension and pressure during underperfusion with NE increased more rapidly in the presence of glyburide. At 60-min underperfusion with NE, the diastolic pressure elevation was still higher in the glyburide treated heart, and decreases in tissue ATP, creatine phosphate (CP), energy charge, phosphorylation potential and CP/inorganic phosphate (P(i)) ratio, and increases in AMP, P(i) and lactate were more marked in the glyburide-treated heart, particularly in the LV subendocardium. Thus, ex vivo glyburide enhanced the increase in LV stiffness and abnormal myocardial energy metabolism during underperfusion with NE in diabetic hearts. These changes were reduced by levcromakalim to the level during underperfusion with NE without glyburide. Insulin did not prevent the glyburide-induced earlier exacerbation of the increase in LV stiffness during underperfusion with NE, but reduced the detrimental effects 20 min after the onset of underperfusion. CONCLUSIONS: KATP channels in the diabetic myocardium probably open during underperfusion with NE, and it helps delay the initiation of the increase in cardiac stiffness. Glyburide may have harmful effects in the ischemic diabetic heart; the myocardial KATP channel blockade during underperfusion with NE enhanced the increase in LV stiffness and abnormal myocardial energy metabolism. The glyburide-induced detrimental effects in the ischemic diabetic heart are prevented by levcromakalim and partly by insulin. PMID- 9349392 TI - The role of L-type Ca2+ current and Na+ current-stimulated Na/Ca exchange in triggering SR calcium release in guinea-pig cardiac ventricular myocytes. AB - OBJECTIVE: This study examines the relative ability of sodium current (INa) stimulated reverse mode Na/Ca exchange and the L-type calcium current (ICa) to trigger calcium-induced calcium release (CICR) in guinea-pig ventricular myocytes. METHODS: Cytosolic Ca2+ transients were recorded from enzymatically dissociated guinea-pig ventricular myocytes using Indo-1. Macroscopic membrane currents were simultaneously recorded using the whole-cell patch-clamp technique. RESULTS: At room temperature (22-25 degrees C) Ca2+ transients were associated with the activation of INa, ICa or INa plus ICa in combination. However, after ICa was blocked by verapamil (10 microM), no Ca2+ transient could be evoked by the activation of INa alone at either -40 or +5 mV. Similar results were obtained with 5 and 8 mM intracellular sodium, and when the temperature of the bathing solution was raised to 35 degrees C and cAMP (10 microM) added to the pipette solution. CONCLUSIONS: From consideration of the relative magnitudes of the Ca2+ influx via ICa and Na/Ca exchange and thermodynamic considerations, we suggest that ICa is the major source of 'trigger' calcium for CICR (and cardiac contraction) under normal conditions. Although the Na/Ca exchanger was incapable of triggering CICR under the conditions of these experiments, we suggest that it may become more important when cytosolic Ca2+ is elevated, a condition which will also lead to decrease the amplitude of ICa. PMID- 9349394 TI - Regional differences in electrical and mechanical properties of myocytes from guinea-pig hearts with mild left ventricular hypertrophy. AB - OBJECTIVE: To investigate electrical and mechanical properties of single myocytes isolated from different regions of the left ventricle in control and hypertrophied hearts. METHODS: Mild cardiac hypertrophy was induced in guinea pigs by aortic constriction. Myocytes were isolated from basal sub-endocardial, basal mid-myocardial and apical sub-epicardial layers of the left ventricle. Action potentials were stimulated at 1 Hz. Membrane currents were measured using the switch-clamp technique. Cell shortening was measured using a photodiode array. RESULTS: In control hearts mean action potential duration (APD) was longer in sub-endocardial myocytes than in sub-epicardial myocytes. In hypertrophy APD was prolonged in sub-epicardial and mid-myocardial myocytes and unchanged in sub endocardial myocytes (APD90 ms, control: sub-endocardial 273 +/- 12, mid myocardial 254 +/- 14, sub-epicardial 229 +/- 9; hypertrophy: sub-endocardial 259 +/- 13, mid-myocardial 291 +/- 9, sub-epicardial 268 +/- 11, P < 0.005, ANOVA). There was no significant regional difference in APD in hypertrophied hearts. In control hearts L-type calcium current (ICa) was similar in all regions. In hypertrophy ICa was increased in sub-epicardial and mid-myocardial myocytes and reduced in sub-endocardial myocytes. Calcium-activated tail currents were not regionally different in control or hypertrophied hearts, but were increased in hypertrophy. CONCLUSIONS: Changes in electrical and mechanical properties associated with hypertrophy are not homogeneous throughout the left ventricle. The difference in APD between sub-endocardial and sub-epicardial myocytes seen in control hearts is lost in hypertrophy. These results may favour the propagation of re-entry arrhythmias in hypertrophied hearts. PMID- 9349395 TI - Characterization and inhibition of beta-adrenergic receptor kinase in intact myocytes. AB - OBJECTIVES: beta-Adrenergic receptor kinase (beta ARK) phosphorylates and thereby inactivates agonist-occupied beta-adrenergic receptors (beta AR). beta ARK is thought to play an important role in the regulation of cardiac function. Therefore, we studied beta ARK activation and its inhibition in intact smooth muscle cells and in cardiomyoblasts. METHODS AND RESULTS: beta AR agonist stimulated translocation of beta ARK was monitored by immunofluorescence labelling with specific antibodies and confocal laser scanning microscopy in DDT MF 2 hamster smooth muscle cells and in H9c2 rat cardiomyoblasts. In unstimulated cells. beta ARK was mainly located in the cytosol. After beta AR agonist stimulation, the beta ARK signal was partially translocated to the membranes. Liposomal gene transfer of the COOH-terminus of beta ARK ('beta ARKmini') as a beta ARK inhibitor led to functional expression of this protein in both cell lines with high efficiency. Western blots with beta ARK antibodies showed a gene concentration-dependent immunoreactivity of the 'beta ARKmini' protein. 'beta ARKmini'-transfected myocytes demonstrated reduced membrane targeting of the beta ARK immuno-fluorescence signal. Additionally, the effect of 'beta ARKmini' on beta AR-induced desensitization of myocytic cAMP accumulation was investigated. In control cells, desensitization with isoproterenol led to a subsequent reduction of beta AR-induced cAMP accumulation. In 'beta ARKmini'-transfected myocytes, this beta AR-induced desensitization was significantly diminished, whereas normal beta AR-induced cAMP accumulation was unaffected. A gene concentration of 2 micrograms 'beta ARKmini' DNA/100,000 cardiomyoblasts, and of 0.7 microgram 'beta ARKmini' DNA/100,000 DDT-MF2 smooth muscle cells led to approximately 5.9- and approximately 5.6-fold overexpressions of 'beta ARKmini' vs. native beta ARK, respectively. These gene doses proved sufficient to attenuate beta-adrenergic desensitization significantly. CONCLUSIONS: (1) beta ARK translocation was evidenced in DDT-MF2 smooth muscle cells and in cardiomyoblasts by confocal laser scanning microscopy. (2) Feasibility of 'beta ARKmini' gene transfer to myocytes was demonstrated, and necessary gene doses for beta ARK inhibition were titered. (3) Overexpression of 'beta ARKmini' functionally interacted with endogenous beta-adrenergic signal transduction, leading to sustained cAMP accumulation after prolonged beta-adrenergic stimulation. PMID- 9349396 TI - Photodynamic therapy inactivates cell-associated basic fibroblast growth factor: a silent way of vascular smooth muscle cell eradication. AB - OBJECTIVE: Procedurally related vascular injury results in a smooth muscle cell (SMC) proliferative response which is in part initiated by SMC release of mitogens, including basic fibroblast growth factor (bFGF). This injury-induced proliferative response is believed to be a key event in intimal hyperplasia development. Photodynamic therapy (PDT), a novel approach found to be effective in inhibiting experimental intimal hyperplasia, produces cytotoxic free radicals resulting in localized SMC eradication. However, this form of SMC injury does not induce an inflammatory or proliferative response in the vessel wall. This study investigated whether PDT-generated free radicals could inactivate cell-associated bFGF normally released with cell injury. METHODS: PDT of bovine SMC was performed in vitro with the photosensitizer CASPc (5 micrograms/ml) and 675 nm laser light using three different fluences: 10, 50, and 100 J/cm2. After PDT, SMC viability was determined with the tetrazolium salt (MTT) assay and cell-associated bFGF was quantitated by ELISA. A SMC mitogenesis assay was utilized to detect cell associated bFGF activity released with SMC injury. RESULTS: In a dose-dependent manner, PDT-generated free radicals reduced cell-associated bFGF levels. After PDT with 100 J/cm2, cell-associated bFGF content was reduced by 88% (P < 0.0002). Of special interest was the finding that PDT with 10 J/cm2 significantly (P < 0.0002) reduced cell viability to around 50%, without affecting cellular bFGF levels. Consequently, a higher PDT dose (100 J/cm2) was needed to significantly (P < 0.001) inhibit the SMC mitogenic response associated with SMC injury. CONCLUSION: These results provide a mechanism to explain how, unlike mechanical or other forms of SMC injury, optimal doses of PDT can locally eradicate medial vascular SMC without resulting in a bFGF-induced initiation of cell proliferation. PMID- 9349397 TI - Block of human cardiac Kv1.5 channels by loratadine: voltage-, time- and use dependent block at concentrations above therapeutic levels. AB - OBJECTIVE: The aim of this study was to analyze the effects of loratadine on a human cardiac K+ channel (hKv1.5) cloned from human ventricle and stably expressed in a mouse cell line. METHODS: Currents were studied using the whole cell configuration of the patch-clamp technique in Ltk- cells transfected with the gene encoding hKv1.5 channels. RESULTS: Loratadine inhibited in a concentration-dependent manner the hKv1.5 current, the apparent affinity being 1.2 +/- 0.2 microM. The blockade increased steeply between -30 and 0 mV which corresponded with the voltage range for channel opening, thus suggesting that the drug binds preferentially to the open state of the channel. The apparent association and dissociation rate constants were (3.6 +/- 0.5) x 10(6).M-1.s-1 and 3.7 +/- 1.6.s-1, respectively. Loratadine, 1 microM, increased the time constant of deactivation of tail currents elicited on return to -40 mV after 500 ms depolarizing pulses to +60 mV from 36.2 +/- 3.4 to 64.9 +/- 3.6 ms (n = 6, P < 0.01), thus inducing a 'crossover' phenomenon. Application of trains of pulses at 1 Hz lead to a progressive increase in the blockade reaching a final value of 48.6 +/- 4.3%. Recovery from loratadine-induced block at -80 mV exhibited a time constant of 743.0 +/- 78.0 ms. Finally, the results of a mathematical stimulation of the effects of loratadine, based on an open-channel block model, reproduced fairly well the main effects of the drug. CONCLUSIONS: The present results demonstrated that loratadine blocked hKv1.5 channels in a concentration-, voltage , time- and use-dependent manner but only at concentrations much higher than therapeutic plasma levels in man. PMID- 9349398 TI - The effects of L-arginine on neointimal formation and vascular function following balloon injury in heritable hyperlipidaemic rabbits. AB - OBJECTIVE: The aims of this study were to determine the morphological and functional consequences of balloon angioplasty of the left subclavian artery of Froxfield heritable hyperlipidaemic (FHHL) rabbits and the influence of oral L arginine therapy on these changes. METHODS: Sixteen-week-old FHHL rabbits were subjected to balloon injury of the left subclavian artery under halothane anaesthesia. Control rabbits (n = 7) were given free access to food and normal tap water. L-Arginine-treated rabbits were given L-arginine (5 g.l-1 in the drinking water for 2 days prior to angioplasty and then for either 2 weeks (n = 7) or 4 weeks (n = 7) after surgery. All rabbits were euthanised 28-30 days after surgery and blood and tissue removed for quantification of neointimal size and determination of endothelial function using isolated vessel tension studies. The ability of the endothelium to prevent platelet aggregation was determined by challenging a vessel ring with carbachol when incorporated into a whole blood sample in which platelet aggregation was induced with collagen. RESULTS: Balloon injury in non-treated rabbits resulted in the development of marked intimal hyperplasia (18.8[3.6]% of the area within the internal elastic lamina) while endothelial function remained intact. Maximum responses to carbachol and calcimycin were, respectively, a 66.6[14.7]% and 46.9[12.9]% relaxation of 5HT induced tone, compared to 58.0[3.2]% and 39.8[9.4]% in non-injured vessels. Maximum contractile responses to 5HT and KCl were unaffected by injury. L Arginine therapy alone had no effect on the vasodilator function of the endothelium, but reduced the endothelium-dependent inhibition of platelet aggregation (68.4[7.8] vs 109[10]% of the maximum extent of platelet aggregation in non-treated and 2-week L-arginine-treated non-injured vessels, respectively). L-Arginine significantly reduced the extent of neointimal formation (7.2[3.9]% of the area within the IEL; P < 0.05 vs. non-treated group). However, L-arginine significantly attenuated the relaxant responses to both carbachol (26.5[10.4]% and 31.4[9.4]% for 2- and 4-week L-arginine groups) and calcimycin (38.7[15.4]% and 16.4[10.7]%) in the injured artery (P < 0.05 compared to non-treated controls). CONCLUSIONS: L-Arginine reduces neointimal formation following balloon catheter injury in heritable hypercholesterolaemic rabbits, which is consistent with previous findings in normocholesterolaemic models. However, in the presence of hypercholesterolaemia, L-arginine has a detrimental effect on endothelial function following injury. This may be a consequence of the presence of lipids in the vascular wall on nitric oxide synthase activity. PMID- 9349399 TI - Down-regulation of endothelin B receptors in autogenous saphenous veins grafted into the arterial circulation. AB - OBJECTIVES: It has been postulated that endothelin (ET) might be involved in the development of atherosclerotic vascular lesions. The present study was done to characterize changes in the contractility and ET receptor subtypes in the autogenous saphenous vein graft (VG). METHODS: The rabbit saphenous vein (SV) was grafted into the ipsilateral femoral artery (FA), and at 4 weeks after the operation, VG was harvested. In the medial layer samples of SV, VG and FA, the cytosolic Ca2+ concentration ([Ca2+]i) and force were monitored using front surface fluorometry of fura-PE3, and mRNA expression of ET receptors was evaluated using the reverse transcription polymerase chain reaction. RESULTS: ET 1 (10(-7) M) developed force in SV, VG and FA, to the same extent. Sarafotoxin (S6c; 10(-7) M) developed force in the SV to the same extent as ET-1. However, S6c did not develop force in FA, and slight force developed in VG. Contractions induced by ET-1 were associated with increases in [Ca2+]i. FA expressed ETA receptor mRNA predominantly, and SV expressed both ETA and ETB receptors mRNAs. In VG, the expression of ETB receptor mRNA was markedly reduced, but expression of ETA receptor mRNA remained unchanged. CONCLUSIONS: Functioning ETB receptors and their mRNA are down-regulated when veins are grafted into the arterial circulation. All these changes in gene expression and function are part of adaptive responses known as 'arterialization'. PMID- 9349400 TI - Nitric oxide causes dysfunction of coronary autoregulation in endotoxemic rats. AB - OBJECTIVE: This study tested the hypothesis that overproduction of endogenous nitric oxide (NO) during endotoxemia may modulate coronary autoregulation and myocardial reactive hyperemia. METHODS: Hearts of endotoxin-pretreated rats and controls were isolated and arranged for perfusion in a Langendorff preparation. Autoregulation was studied by examining flow-pressure relations during stepwise changes in perfusion pressure. The contribution of nitric oxide was examined by perfusion with N omega-nitro-L-arginine (NNLA), an inhibitor of nitric oxide synthesis and methylene blue (MB), an inhibitor of soluble guanylate-cyclase. RESULTS: Endotoxin-treated hearts showed massive coronary vasodilatation and autoregulatory function was impaired at perfusion pressures from 20 to 60 mmHg. Both NNLA and MB reduced coronary flow, improved autoregulation and eliminated differences in coronary flow and autoregulation between the control and endotoxin treated group. Vasoconstriction with vasopressin, a direct smooth muscle constrictor, could not eliminate differences in autoregulation between groups. Reactive hyperemia following coronary occlusion in endotoxin-treated hearts showed decreased duration, flow repayment and repayment ratio. In the presence of NNLA or MB, however, no significant differences in reactive hyperemic flow patterns were present. CONCLUSIONS: These observations suggest that massive coronary vasodilatation due to increased myocardial NO synthesis can result in autoregulatory dysfunction and altered myocardial reactive hyperemia during endotoxemia. PMID- 9349402 TI - The effect of membrane cholesterol content on ion transport processes in plasma membranes. PMID- 9349401 TI - Response of coronary microvascular collaterals to activation of ATP-sensitive K+ channels. AB - OBJECTIVE: Studies have suggested that collateral vessels of the coronary and hind-limb circulations are more sensitive to activation of ATP-sensitive K+ channels than are non-collateral vessels. The objective of the present study was to compare responses of microvascular non-collaterals, native collaterals and stimulated collaterals in the heart to three vasodilators which act through different mechanisms: activation of ATP-sensitive K+ channels with aprikalim, release of nitric oxide with acetylcholine, and endothelium-independent activation of soluble guanylate cyclase with nitroglycerin. METHODS: Collateral growth was stimulated by placing an Ameroid occluder on the proximal left circumflex artery in dogs. Non-collaterals, native collaterals and stimulated collaterals (100-220 microns in diameter) were isolated, cannulated on micropipettes and pressurized in vitro. Vessel diameters were measured using videomicroscopy. RESULTS: Dilation to aprikalim (10(-8)-10(-5) M), acetylcholine (10(-9)-10(-6) M) and nitroglycerin (10(-8)-3 x 10(-4) M) were similar in non collateral, native collateral and stimulated collaterals. Dilation of native collaterals to aprikalim and acetylcholine was attenuated by glibenclamide (10 microM), an inhibitor of ATP-sensitive K+ channels, but not by tetraethylammonium (1 mM), a non-selective inhibitor of K+ channels. Dilation of native collaterals to acetylcholine but not aprikalim was also inhibited by nitro-L-arginine (10 microM), an inhibitor of nitric oxide synthase. CONCLUSION: These findings suggest that microvascular native and stimulated collaterals respond to activation of ATP-sensitive K+ channels and acetylcholine similar to non collaterals of similar size. Thus, changes in reactivity of collaterals to activation of ATP-sensitive K+ channels are not related to changes in the ability of the vessels to respond to vasodilators but may primarily be determined by a change in the distribution of collateral vessel size. PMID- 9349404 TI - The uveoscleral outflow routes. AB - As there is no epithelial barrier between the anterior chamber and the ciliary muscle, aqueous humour may freely pass between the ciliary muscle bundles into the supraciliary and suprachoroidal spaces, from which it is drained through the sclera. This uveoscleral outflow of aqueous humour accounts for 40-60% of the total outflow in monkeys, whereas it is considerably less in (3-8%) in cats and rabbits. Direct measurements in human eyes have suggested that less than 15% is drained by the uveoscleral routes. However, indirect calculations have given a value of about 35% in young adults and 3% in elderly persons (> 60 years). Under normal conditions, in monkeys, the uveoscleral outflow is insensitive to changes in the intraocular pressure, but cyclodialysis and experimental uveitis increase the uveoscleral outflow and make it more pressure sensitive. The uveoscleral outflow is decreased by contraction (pilocarpine) and increased by relaxation (atropine) of the ciliary muscle. Thus, changing the tone of the ciliary muscle may redistribute aqueous humour between the conventional and uveoscleral outflow routes. Prostaglandins decrease the intraocular pressure by increasing the uveoscleral outflow. Two mechanisms seem to contribute to this effect: relaxation of the ciliary muscle and changes in extracellular matrix, causing decreased resistance in the uveoscleral outflow routes. PMID- 9349403 TI - The uvea: questions of pathogenesis and treatment. PMID- 9349405 TI - Genetic influences on sarcoidosis. AB - To investigate the genetic influences underlying the development of sarcoidosis, HLA class II genotyping was performed in Japanese patients with sarcoidosis and healthy controls using the PCR-RFLP method. The frequencies of both DR52 group antigen-associated alleles (HLA-DRB1*11, -DRB1*12 and -DRB1*14) and DRB1*08 alleles were higher in the patient group, suggesting that the common, specific amino acid residue on the DRB1 molecule of these alleles may determine susceptibility to sarcoidosis. Alternatively, it is possible that another susceptibility gene, linked to these DRB1 alleles, exists within the MHC region. We screened the TNFA, TNFB, HSP70-1 and Hum70t genes around the class III region, as well as the HLA-DMA and -DMB genes in the class II region, for genetic polymorphism in sarcoidosis. None of these genes suggested a susceptibility to sarcoidosis. These studies support the thesis that one of the major genetic factors controlling the development of sarcoidosis is located within the DRB1 locus in the HLA class II region. PMID- 9349407 TI - The initiating stimuli for uveitis. PMID- 9349406 TI - What determines the site of inflammation in uveitis and chorioretinitis? PMID- 9349408 TI - Regulation of ocular immune responses. AB - The existence of immune privilege in numerous tissues and organs, including the eye, has been re-emphasised during the past few years, and experimental studies have begun to unravel the multiple mechanisms, both passive and active, that make privilege possible. In this review, recent evidence bearing on the factors responsible for immune privilege in the anterior chamber of the eye is described. One dimension of ocular immune privilege depends upon local factors that limit and modify the expression of immunity. As a consequence, the local expression of immunogenic inflammation is largely curtailed within the eye. Another dimension of ocular immune privilege concerns the modification of induction of systemic immunity to antigens placed within, or arising from, the eye. Systemic responses to ocular antigens are devoid of delayed hypersensitivity T cells and complement fixing antibodies, and the stage for these stereotypic responses is set by factors within the ocular microenvironment acting on indigenous antigen presenting cells. Overall, regulation of immune responses directed at ocular antigens appears to be designed to prevent inflammation from disrupting the visual axis and thereby causing blindness. PMID- 9349409 TI - Antigen presentation in uveitis. AB - Experimental autoimmune uveoretinits (EAU) is not only a valuable model for human inflammatory eye diseases, it is also a useful system for studying many aspects of immunobiology. One such aspect is self/non-self discrimination, the ability of the immune system to tolerate self molecules while responding aggressively to foreign antigens. Our laboratory has used EAU to investigate the mechanisms of T cell tolerance to retinal S-antigen (S-Ag). Several mechanisms have been proposed to maintain T cell tolerance to autoantigens, including clonal deletion and clonal anergy. As immunisation with S-Ag or pathogenic peptides activates uveitogenic T cells, tolerance to this autoantigen cannot be due to clonal deletion. Nevertheless, tolerance acts to keep these existing autoreactive T cells in a naive, or innocuous state. Here we suggest a novel mechanism--low affinity occupancy of the autoantigen-specific T cell receptor (TCR) by self antigen--that may act in concert with the well-known mechanisms to maintain tolerance to S-Ag in the LEW rat. This model differs from clonal anergy in that the missing antigen-presenting cell (APC) activity is not a co-stimulatory function but a TCR co-ligand that increases the avidity of the interaction between the TCR and its peptide-major histocompatibility complex (MHC) ligand. In the absence of this co-ligand only partial signals are generated through the TCR, leading to incomplete T cell activation. This model was deduced from experiments with T cell lines and hybridomas specific for S-Ag, which showed that: (1) autoreactive T cells required a novel APC function, (2) this novel function was necessary to provide complete TCR engagement, and (3) activation of autoreactive T cells was restricted to specific APC. PMID- 9349410 TI - The distribution of immune cells in the uveal tract of the normal eye. AB - Inflammatory and immune-mediated diseases of the eye are not purely the consequence of infiltrating inflammatory cells but may be initiated or propagated by immune cells which are resident or trafficking through the normal eye. The uveal tract in particular is the major site of many such cells, including resident tissue macrophages, dendritic cells and mast cells. This review considers the distribution and location of these and other cells in the iris, ciliary body and choroid in the normal eye. The uveal tract contains rich networks of both resident macrophages and MHC class II+ dendritic cells. The latter appear strategically located to act as sentinels for capturing and sampling blood-borne and intraocular antigens. Large numbers of mast cells are present in the choroid of most species but are virtually absent from the anterior uvea in many laboratory animals; however, the human iris does contain mast cells. Small numbers of what are presumed to be trafficking lymphocytes are present in the uveal tract of normal eyes. There is little data available on the presence or absence of eosinophils. The role of these various cell types in immune homeostasis and ocular inflammation is briefly considered. PMID- 9349411 TI - Antibody response in uveitis. PMID- 9349412 TI - Cytokines: their role in uveal disease. PMID- 9349413 TI - Uveitogenic proteins isolated from bovine iris and ciliary body. AB - Experimental autoimmune anterior uveitis (EAAU) is an animal model for acute anterior uveitis in humans. Previously, we have shown that EAAU can be induced in Lewis rats by a protein(s) associated with ocular melanin. The present study was designed to further purify the pathogenic antigen. Melanin associated antigen (MAA), isolated from bovine iris and ciliary body, was digested with V8 protease and the soluble protein separated on a cation exchange column. The bound protein was eluted with a salt gradient. Lewis rats were immunised with the resulting fractions to test for pathogenicity. Moderate to severe EAAU, with clinical and histopathological features similar to induction with crude soluble bovine MAA, was observed with the 100-200 mM gradient of NH4Cl. Thus, bovine MAA has been partially purified using cation exchange chromatography. Studies are currently under way to purify bovine MAA to homogeneity. PMID- 9349414 TI - T cell mechanisms in experimental autoimmune uveoretinitis: susceptibility is a function of the cytokine response profile. AB - This study addresses the question whether susceptibility versus resistance to experimental autoimmune uveoretinitis (EAU) is connected to a Th1-type (interferon-gamma high, interleukin-4 low), versus a Th2-type (IFN-gamma low, IL 4 high) response. Primed lymph node cells of susceptible Lewis rats produced IFN gamma in response to antigen in culture and transferred EAU to syngeneic recipients, whereas lymph node cells of resistant F344 rats made no IFN-gamma and did not transfer disease. Reversal of the disease pattern, by treatment of F344 rats with B. pertussis toxin and immunisation of Lewis rats with antigen in incomplete Freund's adjuvant, resulted in a parallel reversal of these response patterns. Neither strain produced significant IL-4 responses. A study of the response patterns in mice confirmed that high Th1 responders were susceptible, whereas low Th1 responders and Th2 responders were resistant. We conclude that susceptibility to EAU is connected with a Th1-dominant response, but resistance can involve either a 'null', F344-like response (Th1-low/Th2-low) or a Th2 dominant response. PMID- 9349415 TI - Mechanisms of inflammatory response in sympathetic ophthalmia and VKH syndrome. AB - Although the inciting events in the pathogenesis of sympathetic ophthalmia and Vogt-Koyanagi-Harada (VKH) syndrome are different, these two forms of bilateral, granulomatous uveitis share several clinical, histopathological and immunohistochemical features, including their association with HLA types and in their in vitro T-cell response to retinal antigens. These clinical and immunopathological features indicate that there is an underlying T-cell mediated autoimmunity to uveal/retinal antigens in the development of these forms of uveitis. Both forms exhibit preservation of the choriocapillaris and retina despite extensive inflammatory cell infiltration in the choroid. Recent experimental studies suggest that this preservation of choriocapillaris could be the result of anti-inflammatory products secreted by the retinal pigment epithelium, including transforming growth factor-beta and a novel protein called retinal pigment epithelial protective protein that is known to suppress the phagocyte generation of superoxide. Such suppression of the oxidant release in the choroidal inflammation could help protect the uvea from necrotic change and preserve the choriocapillaris from inflammatory cell infiltration. PMID- 9349416 TI - Perspectives on gene therapy in the treatment of ocular inflammation. AB - Gene therapy may become a powerful therapeutic strategy. However, the application of this method in the treatment of ocular disease presents us with interesting and unique questions. Gene therapy for ocular inflammatory disease has the potential for both therapeutic interventions and a method for studying mechanism of disease. An evolving philosophy on this subject would support the use of somatic gene therapy for ocular inflammatory disease, even if not life threatening. Major technical questions remain, including the use of the appropriate vector, the best methodology for the stable insertion into the genome, and the duration and intensity of expression of the transgene. Various transgenes encoding a wide variety of proteins can be envisaged for the insertion of genes. The study of gyrate atrophy, an hereditary ocular disorder and an excellent candidate for gene therapy, has given us enormous information in the development of practical therapeutic strategies, as have in vitro studies of gene insertion. Future concerns will need to concentrate on the use of better methods for gene insertion and homologous recombination techniques for the development of animal models and later as a strategy for gene therapy. The use of gene therapy as a drug delivery system must also be considered. In addition, the elucidation of the various events controlling transcription for the expression of transgenes in various resident ocular cells is necessary. PMID- 9349417 TI - New therapeutic options in uveitis. AB - Most patients with sight-threatening posterior uveitis eventually end up on systemic medication to control their disease. Although the more aggressive approach to the use of these drugs does offer the patient a better chance of significant visual improvement at least in the short term, this is often associated with severe systemic side-effects in both the young and older patient. Cyclosporin has become a very useful second-line agent as a steroid sparer in those patients who can tolerate it. However, it is not suitable for or effective in everyone and the other currently available drugs are often of limited effectivity or associated with major systemic sequelae. This paper summarises the therapeutic approaches currently being examined to define whether they have a role in the better management of these patients in the future. Particularly exciting is the potential for sustained intraocular drug delivery so that adequate drug levels are achieved inside the eye without the necessity for systemic administration. PMID- 9349418 TI - The microcirculation of choroidal and ciliary body melanomas. AB - The microcirculation of ciliary body and choroidal melanomas is remodelled into patterns. The presence of microvascular networks, composed of back-to-back loops that encircle microdomains of tumour, and parallel vessels with cross-linking, are associated with death from metastatic melanoma. The formation of these complex vascular patterns may result from reciprocal interactions between the tumour cell and the extracellular matrix, and pattern formation may reflect an invasive tumour cell phenotype. Ciliary body and choroidal melanomas are among the few forms of cancer treated before a pathologist assigns a grade to indicate whether tumour is likely to follow a benign or aggressive course. There is evidence to suggest that prognostically significant microcirculatory patterns may be detectable by non-invasive imaging techniques that may provide a substitute for biopsy to guide the clinical management of patients with these sight- and life-threatening tumours. PMID- 9349419 TI - Uveal melanoma: tumour phenotype and metastatic potential. PMID- 9349420 TI - Genes coding for melanoma antigens recognised by cytolytic T lymphocytes. AB - It is now well established that human melanoma cells express antigens that are recognised by cytolytic T lymphocytes derived from the tumour-bearing patient. The molecular definition of these antigens is progressing at an accelerated pace. The currently characterised melanoma antigens can be classified into three categories: differentiation antigens, antigens encoded by genes that are specifically expressed in tumours, and antigens encoded by mutated genes. Several of these antigens are sufficiently tumour-specific to qualify them as candidate anti-cancer vaccines in melanoma patients. PMID- 9349421 TI - Immunoregulation of intraocular tumours. AB - Intraocular tumours reside within an organ that provides sanctuary from many immunological defence mechanisms. Antigens displayed on many intraocular tumours can elicit an aberrant systemic immune response in which systemic antigen specific delayed-type hypersensitivity (DTH) is actively downregulated, thus denying the host one potential effector mechanism for controlling its intraocular tumour. Constituents within the aqueous humour inhibit the expression of DTH and natural killer cell effector mechanisms within the eye and thus protect intraocular tumours from immune-mediated rejection. Some experimental intraocular tumours in mice express potent tumour-specific antigens that stimulate the expansion of tumour-specific robust cytotoxic T lymphocyte (CTL) populations which enter the eye and mediate tumour rejection by piecemeal tumour necrosis. However, the presence of tumour-infiltrating lymphocytes (TIL) within an intraocular tumour does not inevitably lead to tumour resolution. In some cases CTL precursors infiltrate the intraocular tumour but fail to differentiate into mature cytolytic effector cells. Although uveal melanomas express melanoma specific and melanoma-associated antigens that are capable of eliciting both humoral and cellular immunity, formidable barriers prevent the expression of tumour immunity. These barriers include: (a) anterior chamber-associated immune deviation; (b) in situ suppression of DTH effector cells; (c) suppression of natural killer cell activity in oculi; and (d) inactivation of the complement cascade by regulatory proteins expressed on uveal melanoma cells. PMID- 9349422 TI - The Ashton Lecture. Uveal melanoma: the past, the present and the future. PMID- 9349423 TI - Bridging the 'commercialisation gap' in Europe. PMID- 9349424 TI - Foetal gene delivery in mice by intra-amniotic administration of retroviral producer cells and adenovirus. AB - With the aim of developing foetal gene therapy for cystic fibrosis, we have investigated the possibility of gene targeting to the mouse foetus with two different viral vector systems and at different times of gestation. We report here that recombinant retrovirus producing cells administered into the intra amniotic cavity of mid- to late-gestation mouse MF1 foetuses survive in the amniotic fluid and are able to engraft to a certain extent in foetal tissues. By production of infectious virus they mediate transduction and beta-galactosidase transgene expression in neighbouring foetal tissues 24 to 72 h following injection. Retrovirus producer cells could, therefore, become a means to overcome the limitations of low retroviral titre, for in vivo foetal gene transfer. To investigate the developmental stage at which transduction of the airways and enteral systems can be obtained we also administered a highly infective first generation adenoviral vector (AdRSV beta gal) into the amniotic cavity of foetal mice between 13 to 16 days post coitus, beta-galactosidase activity was detected between 24 to 120 h after injection. The highest levels of transgene expression were generally observed between 48 to 72 h following injection of the adenoviral vector. We demonstrate that infection of the pulmonary airways is dependent on the developmental stage of the foetus and can be achieved on the 15th day of gestation. PMID- 9349425 TI - In vivo gene transfer via intravenous administration of cationic lipid-protamine DNA (LPD) complexes. AB - A novel LPD formulation has been developed for in vivo gene transfer. It involves the interaction of plasmid DNA with protamine sulfate, a cationic polypeptide, followed by the addition of DOTAP cationic liposomes. Compared with DOTAP/DNA complexes, LPD offers better protection of plasmid DNA against enzymatic digestion and gives consistently higher gene expression in mice via tail vein injection. When a luciferase reporter gene was employed, gene expression was found in all tissues examined including lung, heart, spleen, liver and kidney with the highest expression in the lung. The in vivo efficiency of LPD was dependent upon charge ratio and was also affected by the lipid used. Increasing the amount of DNA delivered induced an increase in gene expression. The optimal dose was approximately 50 micrograms per mouse at which concentration approximately 20 ng luciferase protein per milligram extracted tissue protein could be detected in the lung. Increasing the DNA to 100 micrograms per mouse resulted in toxicity and death of the animal. Gene expression in the lung was detected as early as 1 h after injection, peaked at 6 h and declined thereafter. High expression was also found in the spleen 6 h after injection but dropped very rapidly thereafter. The in vivo gene expression by LPD was dependent upon the route of administration since intraportal injection of LPD led to about a 100 fold decrease in gene expression in the lung as compared with i.v. injection. Using lacZ as a reporter gene, it was shown that endothelial cells were the primary locus of transgene expression in both the lung and spleen. No sign of inflammation in these organs was noticed. Since protamine sulfate has been proven to be nontoxic and only weakly immunogenic in humans, this novel vector may be useful for clinical gene therapy. PMID- 9349426 TI - Tetracycline-responsive gene expression in mouse brain after amplicon-mediated gene transfer. AB - Amplicon vectors incorporate genetic elements from Herpes simplex virus (HSV) in a plasmid form which is packaged into virions in the presence of a replication defective helper virus. We constructed a new amplicon vector, pHermes-tet-lacZ, that carries the bacterial beta-galactosidase (lacZ) gene under the control of a minimal promoter preceded by a heptameric tetracycline operator. The minimal promoter element is activated by a tetracycline-responsive hybrid protein, the gene for which is also present in the vector. This amplicon was propagated in parallel in two different permissive cell lines, E5 and 2-2, in the presence of two helper viruses, d120 and 5dl1.2, respectively. The viral stocks produced were injected into the hippocampal region of the mouse brain, where strong localized expression of the transgene developed in the granular cell layer of the dentate gyrus with limited cytotoxicity. The transgene expression could be repressed by a factor of 10 after administration of tetracyclines. The repression level depended on the helper virus present in the injected viral stock. The in vivo regulation of transgene expression conferred by the tetracycline responsive element improves the flexibility of amplicon vectors as tools for gene transfer into the brain. PMID- 9349427 TI - Retinoids augment the bystander effect in vitro and in vivo in herpes simplex virus thymidine kinase/ganciclovir-mediated gene therapy. AB - Metabolic cooperation via gap junctional intercellular communication (GJIC) is an important mechanism of the bystander effect in gene therapy using the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) 'prodrug' system. Since retinoids have been reported to increase GJIC by induction of connexin expression, we hypothesized that these compounds could be used to augment the HSVtk/GCV bystander effect. Addition of all-trans retinoic acid increased GJIC in tumor cell lines, augmented expression of connexin 43, and was associated with more efficient GCV-induced in vitro bystander killing in cells transduced with HSVtk via either retrovirus or adenovirus vectors. This augmentation of bystander effect could also be seen in vivo. HSVtk-transduced tumors in mice treated with the combination of GCV and retinoids were significantly smaller than those treated with GCV or retinoids alone. These results provide evidence that retinoids can augment the efficiency of cell killing with the HSVtk/GCV system by enhancing bystander effects and may thus be a promising new approach to improve responses in gene therapy utilizing the HSVtk/GCV system to treat tumors or vascular restenosis. PMID- 9349428 TI - Interaction of vesicular stomatitis virus-G pseudotyped retrovirus with CD34+ and CD34+ CD38- hematopoietic progenitor cells. AB - Retroviral vectors have had limited success in mediating gene transfer to hematopoietic stem cells, particularly in primates, due in part to low or absent expression of the amphotropic receptor (RAM-1). We have been interested in determining whether retrovirus pseudotyped with vesicular stomatitis virus G protein (VSV-G) would allow more efficient gene delivery to hematopoietic stem cells as the VSV-G receptors appear to be ubiquitously present phospholipids. However, we previously found that completion of retroviral vector reverse transcription does not occur in CD34+ CD38- hematopoietic stem cells that were exposed to VSV-G pseudotyped retrovirus. To determine at which stage the block to infection of CD34+ CD38- cells occurs, we confirmed by FACS analysis that VSV-G pseudotyped viral particles could bind to CD34+ CD38- cells. Virus binding to CD34+ cells was saturable at 4 degrees C but nonsaturable at 37 degrees C, up to a multiplicity of infection of 1080. This suggests that surface levels of phospholipid receptors available for viral binding are limiting on CD34+ cells. Cytokine stimulation increased virus binding to CD34+ cells. However, no increase in the level of surface phosphatidylserine (PS), a strong candidate for the VSV-G receptor, was seen as detected by the PS-specific reagent, annexin V. This suggests that another molecule is serving as the VSV-G receptor on CD34+ cells. Here, we show that once virus binding to cytokine-stimulated CD34+ CD38- cells has occurred, virus fusion proceeds efficiently as determined by octadecyl rhodamine (R18) fusion assays. Taken together with our previous observation that reverse transcription does not occur in CD34+ CD38- cells, we suggest that there are intracellular mechanisms leading to blockage of complete reverse transcription of the retrovirus in CD34+ CD38- cells. This has important implications for retrovirus-mediated gene transfer to quiescent stem cells. PMID- 9349429 TI - Enhanced maintenance and retroviral transduction of primitive hematopoietic progenitor cells using a novel three-dimensional culture system. AB - Current techniques for the in vitro maintenance of hematopoietic stem cells often lead to loss of pluripotency. Overcoming the technical difficulties that result in alterations in stem cells in vitro has important implications for areas of basic science and clinical medicine such as cell expansion, bone marrow transplantation and gene therapy. Recent insights into hematopoietic stem cell biology have demonstrated that the three-dimensional architecture of the culture environment may influence the maintenance of stem cell pluripotency in vitro. An intriguing hypothesis is that the utilization of three-dimensional culture systems may improve the maintenance and manipulation of these cells in vitro. We report that a novel, three-dimensional, tantalum-coated porous biomaterial (TCPB) may be employed effectively as a hematopoietic progenitor cell culture device that offers distinct advantages over conventional culture systems. Specifically, we demonstrate that the use of TCPB for culturing hematopoietic progenitor cells in the absence of exogenous cytokines results in enhanced hematopoietic progenitor cell survival, improved maintenance of the immature CD34+/38- phenotype, and improved retroviral transduction of CD34+ cells and long-term culture initiating cells (LTCIC), without compromising multipotency, as compared with cultures in plastic dishes or bone marrow stroma. These findings suggest that this three-dimensional culture system may be useful in advancing the in vitro culture and transduction of hematopoietic stem and progenitor cells. PMID- 9349430 TI - Safety of a single aerosol administration of escalating doses of the cationic lipid GL-67/DOPE/DMPE-PEG5000 formulation to the lungs of normal volunteers. AB - Several groups are assessing the use of cationic lipids for respiratory gene therapy. To date no human data are available regarding the safety of intra pulmonary cationic lipid delivery. In preparation for a trial of pulmonary delivery of the CFTR gene, we have assessed the safety of nebulised lipid GL 67/DOPE/DMPE-PEG5000 (GL-67A), the cationic lipid formulation to be used in this study. Fifteen healthy volunteers were given incremental doses of GL-67A via a Pari LC Jet nebuliser; three volunteers in each of five dosing cohorts with a week interval between cohorts. Markers of safety included clinical assessment, measurement of lung function, chest CT scan, serological testing and analysis of induced sputum. Measurements were taken before administration and at intervals up to 21 days thereafter. No adverse clinical events were seen or any statistically significant changes in spirometry or gas transfer. There were no clinically significant changes in any of the blood parameters and no CT changes were seen. Comparisons of the cellular subpopulations (neutrophils, eosinophils, lymphocytes and macrophages) in induced sputum showed no significant alterations following administration of the GL-67A. This study suggests that a single application of aerosol formulation of GL-67A does not result in clinically detectable changes when given by nebulisation into the lungs of normal volunteers and provides an indication of a lipid dose tolerated in man. PMID- 9349431 TI - Adenovirus-mediated transduction of ribozymes abrogates HER-2/neu and pleiotrophin expression and inhibits tumor cell proliferation. AB - The combination of specific gene targeting technologies with efficient gene delivery systems could provide the means to evaluate the concept of anticancer strategies designed to block expression of potentially rate-limiting tumor promoting factors. Here, we constructed adenoviruses expressing hammerhead ribozymes targeted to two of these factors, the tyrosine kinase receptor HER 2/neu or the growth factor pleiotrophin (PTN). Adenovirus-mediated transduction of either HER-2/neu- or PTN-targeted ribozymes depleted the respective RNAs and inhibited protein expression significantly in three different human cancer cell lines. This resulted in almost complete abrogation of HER-2/neu- or PTN-dependent cancer-cell proliferation, thus demonstrating the feasibility of this approach as a future cancer gene therapy. PMID- 9349432 TI - Overcoming the inhibitory effect of serum on lipofection by increasing the charge ratio of cationic liposome to DNA. AB - Since cationic liposome was first developed as a lipofection reagent, a drawback has been noted in that the efficiency of lipofection decreases dramatically after addition of serum to the lipofection medium. This drawback hampers the application of cationic liposome for systematic delivery of genes. In the present studies, we found that the effect of serum on DC-chol liposome-mediated lipofection is dependent on the charge ratio of liposome to DNA. Serum inhibited lipofection activity of the lipoplex at low charge ratios, whereas it enhanced the lipofection activity at high charge ratios. This phenomenon was observed using DOTAP/DOPE but not lipofectamine. Measurement of cellular association of DNA showed that serum could reduce the binding of lipoplex to cells at all tested charge ratios, i.e. 0-10.6. Removal of negatively charged proteins from serum by DEAE Sephacel column abolished the inhibitory effect of serum on lipofection. The fraction contained only negatively charged serum proteins which strongly inhibited lipofection at low charge ratios but not at higher charge ratios. Furthermore, preincubation of serum with positively charged polylysine, which neutralized negatively charged serum proteins, eliminated the inhibitory effect of serum on lipofection. In summary, inactivation of cationic liposome by serum is due to negatively charged serum proteins and it can be overcome by increasing charge ratio of cationic liposome-DNA lipoplexes or by neutralizing the serum with polylysine. PMID- 9349433 TI - Protamine sulfate enhances lipid-mediated gene transfer. AB - A polycationic peptide, protamine sulfate, USP, has been shown to be able to condense plasmid DNA efficiently for delivery into several different types of cells in vitro by several different types of cationic liposomes. The monovalent cationic liposomal formulations (DC-Chol and lipofectin) exhibited increased transfection activities comparable to that seen with the multivalent cationic liposome formulation, lipofectamine. This suggests that lipofectamine's superior in vitro activity arises from its ability to condense DNA efficiently and that protamine's primary role is that of a condensation agent, although it also possesses several amino acid sequences resembling that of a nuclear localization signal. While the use of polycations to condense DNA has been previously reported, the of protamine sulfate, USP as a condensation agent was found to be superior to poly-L-lysine as well as to various other types of protamine. These differences among various salt forms of protamine appear to be attributable to structural differences between the protamines and not due to differences in the net charge of the molecule. The appearance of lysine residues within the protamine molecule correlate with a reduction in binding affinity to plasmid DNA as well as an observed loss in transfection enhancing activity. This finding sheds light on the structural requirements of condensation agents for use in gene transfer protocols. Furthermore, protamine sulfate, USP is an FDA-approved compound with a documented safety profile and could be readily used as an adjuvant to a human gene therapy protocol. PMID- 9349434 TI - Induction of antibody response to human tumor antigens by gene therapy using a fusigenic viral liposome vaccine. AB - Development of effective cancer vaccines would help prevent and control tumor progression. A novel approach of immunizing against tumor antigens is in vivo gene vaccination. We have developed a fusigenic viral liposome vector using HVJ (hemagglutinating virus of Japan) and liposome to deliver human tumor antigen genes effectively to cells in vivo. Plasmids containing the human tumor antigen genes MAGE-1 and MAGE-3 were encapsulated in fusigenic viral liposomes and injected into mice intramuscularly. MAGE-1 and -3 recombinant proteins were used in Western blotting and affinity ELISA for assessment of antibody responses. Mice immunized with MAGE-1 and -3 gene vaccine individually were shown to produce anti MAGE-1 and -3 IgG antibody responses respectively. Animals immunized with plasmid alone did not induce anti-MAGE-1 or -3 IgG responses. Antibody responses could be enhanced on reimmunization with the gene vaccines. Muscle biopsies taken after vaccine injection were verified to express gene-specific mRNA transcripts. Mice immunized with MAGE-1 or -3 gene vaccines were shown to induce antibodies that could cross-react with the respective recombinant proteins. This study demonstrates that in vivo immunization using HVJ-liposome containing human tumor antigen genes can effectively deliver and induce immune responses to the respective whole proteins. PMID- 9349435 TI - Direct retrovirus-mediated gene transfer to the synovium of the rabbit knee: implications for arthritis gene therapy. AB - We have investigated the feasibility of using high-titer murine leukemia virus based retroviral vectors to deliver exogenous genes to naive and chronically inflamed knee joints of rabbits in vivo. Intraarticular injection of retrovirus encoding beta-galactosidase (beta-gal or lacZ) was found to transduce synoviocytes in both naive and inflamed joints, but a significantly higher number of lacZ+ cells were found in inflamed knees. Using a retrovirus encoding a secretable marker, human growth hormone (hGH), quantitative comparison of ex vivo and in vivo gene delivery methods demonstrated that transgene expression following in vivo gene transfer was at least equivalent to that of the ex vivo method in inflamed knees. In addition, hGH transgene expression was maintained for at least 4 weeks. These experiments suggest that high-titer retroviral vector could be used for efficient in vivo gene transfer to inflamed joints in patients with rheumatoid arthritis (RA). PMID- 9349436 TI - Efficient in vivo delivery of DNA to pulmonary cells using the novel lipid EDMPC. AB - We compared the efficacy of gene transfer in vitro and in vivo using various formulations of DNA-lipid complexes based on the novel cationic lipid EDMPC (1,2 dimyristoylsn-glycero-3-ethylphosphocholine, chloride salt). In vitro studies analyzed delivery of marker genes to four established cell lines, including two of pulmonary origin. The in vivo analysis used intralobar delivery of marker genes and CFTR to mice and rats. We observed a lack of positive correlation between those DNA-EDMPC formulations that delivered DNA most efficiently in vitro and those that worked best in vivo. Intralobar DNA delivery to rodents mediated by EDMPC was efficient. The high level of gene delivery by DNA-EDMPC formulations demonstrates that efficient lipid-mediated gene transfer to the lung is possible. PMID- 9349437 TI - Delay in resumption of the activity of tetracycline-regulatable promoter following removal of tetracycline analogues. AB - The tetracycline-regulatable system (TRS) has become a widely adopted tool for modification of gene expression and analysis of gene function in mammalian cells, plants and transgenic animals. We have studied the potential application of the TRS in gene therapy, using a single vector containing both the tetracycline controlled transactivator (tTA) and the tTA-responsive promoter (tRP) transcribing mouse GM-CSF. Stable 293 cells established using this vector were used to study the kinetics of the TRS in response to various tetracycline analogues. Dose-response studies show that doxycycline is the most potent analogue in abolishing tTA activity. Kinetic studies indicate that, at 1,000 ng/ml, all the analogues have similar efficiencies in down-regulating the system in given time. In contrast, following the removal of the analogues, there is a temporal, dose-dependent delay in resumption of the tRP activity. The time taken for resumption of near-optimal tRP activity is approximately 48 h for tetracycline, 144 h for anhydrotetracycline, 192 h for minocycline and 216 h for doxycycline when cells were pretreated with 1000 ng/ml of these antibiotics. This property of the analogues can be employed in planning a desired course of transgene regulation. PMID- 9349438 TI - Nutritional evaluation of some commercial baby foods consumed in Saudi Arabia. AB - The nutritive value of six baby foods based on milk (Nido, Wadi Fatima and Gain) and milk-cereal blends (Cerelac with wheat, Cerelac with rice and Milupa 2) commonly used in the Kingdom of Saudia Arabia was evaluated chemically including fatty acids analysis and biologically in growing rats. The milk based products vs milk-cereal blends provided (per 100 kcal) protein (3.8-5.0 g vs 3.7-3.8 g), fat (5.2-5.7 g vs 2.0-4.8 g), available carbohydrates (7.3-9.5 g vs 10.5-16.6 g), Ca (159-189 mg vs 101-145 mg), Mg (15-18 mg vs 14-20 mg), Na (32-39 mg vs 42-51 mg), K (160-180 mg vs 122-144 mg), Fe (1.4-1.8 mg vs 1.5-1.9 mg), Cu (0.04-0.09 mg vs 0.09-0.1 mg), Zn (0.8-1.2 mg vs 0.8-1.1 mg), and linoleic acid (208-1343 mg vs 518-639 mg). Metabolizable energy (ME) values in milk based products (487-495 kcal/100 g) were higher than milk-cereal blends (404-473 kcal/100 g). The true protein digestibility (TD) varied from (93-95%) in milk based foods to (94-95%) in milk-cereal blends. The net protein utilization (NPU) ranged between (0.74 0.78) in milk based products and (0.68-0.74) in milk-cereal blends. The net dietary protein calorie percent (NDP cal%) was higher in milk based foods (11.7 15.0%) than milk-cereal blends (10.2-11.1%). An imbalance of calories and nutrients in some baby foods was noticed. However, the protein quality was satisfactory and could meet the protein requirements of infants and toddlers as indicated by NDP cal% values. PMID- 9349439 TI - Effects of debittering on grapefruit juice acceptance. AB - This study was conducted to assess the acceptance of grapefruit juice which has undergone a debittering process. The sensory effect of debittering and the sensory attributes of sourness, sweetness, bitterness, and aftertaste were appraised, and the correlation between chemical and sensory analyses of the debittered juice were identified. The effect of added grapefruit flavor on perception of sweetness and sourness was statistically significant. Both the level of bitterness and storage duration of grapefruit were shown to influence the way judges perceived bitterness and sweetness. Storage study showed no difference in aftertaste, which may increase consumers buying interest in debittered juice with a high level of bitterness (450 ppm). PMID- 9349441 TI - Biodeterioration potentials of fungal isolates from vegetable oils. AB - The biodeterioration potentials of fungi isolated from vegetable oils were investigated. Growth of pure cultures of some mould species (Aspergillus flavus, Aspergillus niger, Aspergillus sp., Penicillium sp. and Fusarium sp.) and a mixed yeast culture (Saccharomyces sp., Candida sp. and Hansenula sp.) were monitored using six vegetable oils as substrates. Mould growth resulted in an increase in mycelial dry weight (g/100 ml) of the cultures in all the oils. Yeast growth in the oils caused decrease in pH, increase in optical density and increase in total viable count (TVC) in all the oils. Gas chromatographic analysis of the heptane extracts of the oils revealed that inoculated and uninoculated oil samples displayed similar chromatograms. Yeast growth in the oils also resulted in loss of some fatty acid components as well as biosyntheses of new ones. PMID- 9349440 TI - Functional properties of some varieties of African yam bean (Sphenostylis stenocarpa) flour--III. AB - The proximate analysis, determination of nutritionally valuable minerals and the functional properties of the seed flour of African yam bean (AYB) (Sphenostylis stenocarpa) were investigated. Three different colour varieties of whole seeds and the dehulled samples from the same source were identified and processed for the study. The average composition of the whole seeds was as follows: 20.50% protein, 8.25% fat, 59.72% total carbohydrate, 3.26% total ash and 8.10% moisture, while the corresponding dehulled samples contained 21.9% protein, 8.63% fat, 60.89% total carbohydrate, 2.20% ash, and 6.35% moisture. The whole seeds were rich in potassium (649.49 mg/100 g) and phosphorus (241.21 mg/100 g) while values for dehulled samples were 471.35 mg/100 mg/100 g and 245.81 mg/100 g respectively. The seed flours have good gelation property, protein solubility varied with pH with high solubilities in acid and alkali. Dehulled samples had lower values for fat absorption capacity and foaming capacity but higher values for water absorption capacity, foaming stability rate change and fat emulsion stability rate change. PMID- 9349442 TI - Effect of food processing treatments on generation of resistant starch. AB - The resistant starch (RS) contents of rice and waxy amaranth starch subjected to processing treatments like cooking by boiling, pressure cooking, roasting, extrusion cooking, frying and drum drying have been reported. Compared to the contents in the native starch, when expressed on dry wt basis, in both cases, cooking by boiling and pressure cooking resulted in increase in RS content whereas all other processing treatments resulted in a decrease in RS contents. PMID- 9349443 TI - Low-fat papadams from black gram-tapioca blends. AB - Papadams, made from black gram (Phaseolus mungo) and largely manufactured on cottage scale are popular in the Indian dietary. Escalating prices of black gram coupled with abundant availability of tapioca flour at almost 1/8th the price of black gram prompted the study of papadam characteristics using blends of black gram and tapioca flour. Tapioca flour up to 25% substitution did not alter the sensory attributes. However, the expansibility of the product decreased on addition of tapioca flour; a fact which could be overcome by using carboxymethyl cellulose (CMC) at 3% level of black gram flour. This decreased the oil content to 19.7% and also increased the expansibility to 5.539%. PMID- 9349444 TI - Prevalence of anaemia in pregnant women during the last trimester. AB - Anaemia is a very common condition during pregnancy. This is particularly so in developing countries where the level of intake of iron rich foods is low; malaria and other intestinal parasites are common. This study was conducted to determine the prevalence of anaemia and the type of anaemia existing in pregnant women in Morogoro municipality. The effect of anaemia on infant birth weight was also examined. Twenty randomly selected pregnant women in their last trimester of pregnancy were studied. The subjects were recruited from the three maternal and child health clinics in the municipality. The subjects were not taking iron, folate or vitamin B12 supplements at the time of the study. Blood samples were collected from subjects during their routine visit to maternal and child health clinics. A series of determinations was conducted to determine haemoglobin concentration (Hb); packed cell volume (PCV); red blood cells count (RBC); serum iron (SI); and total iron binding capacity (TIBC). The effect of anaemia on the weight of new born babies was examined by calculating the correlation coefficient of birth weight and haematological indexes. The mean values (SD) of haematological indexes were as follows: Hb 8.7 +/- 1.5 g/dl; PCV 30.4 +/- 5.1%; RBC 2.5 +/- 0.6 x 10(2)/l; mean corpuscular haemoglobin concentration (MCHC) 28.9 +/- 4 g/dl; mean corpuscular volume (MCV) 151.5 +/- 120 fl and mean corpuscular haemoglobin (MCH) 35.2 +/- 7.9 pg. The results have shown that 95% of the subjects were anaemic at the time of the study. All subjects were suffering from iron, folate and vitamin B 12 deficiencies. This suggests that all subjects had a combination of microcytic and megaloblastic anaemia. The results have also shown that there was a positive correlation (r = 0.76; P = 0.01) between Hb concentration and weight of the infants at birth. Subjects who had Hb concentration of below 7.4 g/dl delivered infants that were weighing below 2500 g (mean birth weight of 2160 +/- 228 g). For those who had an Hb concentration of above 9.5 g/dl delivered infants weighing more than 3000 g (mean 3142 +/- 329 g). The mean birth weight of the infants born to anaemic subjects (Hb < 7.9 g/dl) was significantly lower compared to that of infants born to non-anaemic subjects. This observation suggests that anaemia had a significant influence on the birth weight of the infant. This could also be an indication of poor food security in general. Major causes of anaemia were identified as being poor dietary intake of iron rich foods and probably poor utilisation due to diseases such as malaria. All women had basic knowledge on anaemia. Most of the information was obtained from maternal and child health clinics (76%), schools (15%) and radio programmes (4%). However, despite their awareness on anaemia, the women were still anaemic. The main reason was lack of economic access to appropriate foods. PMID- 9349445 TI - The female Spanish population: a group at risk of nutritional iron deficiency. AB - Iron deficiency is one of the most common nutritional problems in the world. It is frequent in both developed and developing countries and mainly affects women of childbearing age. The aim of the present study was to investigate the prevalence of iron deficiency in a group of young women from Madrid, Spain. The study subjects were a group of 130 women aged between 19 and 35 (24.53 +/- 0.24 years). Measurements were made of iron intake and also of the haematological and biochemical indicators of iron status. 10.7% of subjects showed iron deficiency (defined as the recording of at least two indicator parameters with values below normal). The high incidence of iron deficiency at blood level (10.7%) coincided with the low iron intake of these subjects (11.08 +/- 2.98 mg/day). 98.3% of subjects showed intakes below recommended. Observed intake covered only 61.6% of recommended intake. 3.9% of subjects presented ferropenic anaemia, i.e. they showed both iron deficiency and low haemoglobin levels. PMID- 9349446 TI - The fate of fat in the infant's colon. PMID- 9349447 TI - Molecular mechanisms of inherited insulin resistance. PMID- 9349448 TI - Lactic acidosis and oxygen debt in African children with severe anaemia. AB - A syndrome of severe anaemia (Hb < or = 5 g/dl), particularly severe malarial anaemia (SMA), remains a major cause of childhood mortality in sub-Saharan Africa. We hypothesized that the lactic acidosis which identifies those at the greatest risk of death often represents an oxygen debt incurred as a result of inadequate tissue perfusion. To examine this hypothesis, we measured oxygen consumption (VO2) using a portable metabolic monitor. Blood lactate and acid-base status were also determined. Pre-transfusion data on 44 children (28 with mild symptoms, 7 with respiratory distress and 9 controls) demonstrated very close dependence of VO2 on body surface area (BSA, R2 = 0.86, p < 0.001). After correcting for BSA, no significant differences were observed in mean VO2 values of the three clinical groups, indicating that a critical reduction in oxygen delivery is not the sole explanation for the development of a lactic acidosis and severe symptoms. Nine children (including five of the original 44) were monitored during transfusion. In four of the five with SMA, severe symptoms and severe lactic acidosis, transfusion produced a marked, transient increase in VO2 (maximum 30-41%), with a marked fall in blood lactate and clinical improvement. These data suggest that some children with SMA and respiratory distress accumulate an oxygen debt when a relatively high oxygen demand outstrips supply, this debt being repaid when supply is increased during transfusion. However, in the remaining one of these five children, an increase in VO2 (maximum 20%), was accompanied by a rise in blood lactate and clinical deterioration, suggesting that more pathophysiologically complex mechanisms, which may predominate in some children. PMID- 9349449 TI - Malignant hypertension in young women is related to previous hypertension in pregnancy, not oral contraception. AB - Previous studies have suggested that one-third of women of childbearing age who develop malignant phase hypertension (MHT) are likely to be taking oral contraceptives (OC). We surveyed 104 women with a history of MHT. None of the 65 aged > 45 years were taking OC or other sex hormones. Thirty-nine (mean age 34.9 years, SD 8.0) were aged 15-44 years at presentation: 22 Caucasian, 10 Black/Afro Caribbean and seven Indo-Asian. Of these 39, 22 had a history of hypertension in pregnancy (group 1), and 17 did not (group 2). Three of group 1 also had a history of OC-induced hypertension. None were pregnant, but one was taking an OC at presentation with MHT. Blood pressures at presentation and follow-up, and mean serum urea and creatinine at presentation were similar between groups, as was median survival (96 vs. 47 months, Lee-Desu statistic 0.75, p = 0.38). There was a trend towards poorer renal function at follow-up in group 1 patients, with higher mean serum urea and creatinine levels. The causes of death were renal failure (5), stroke (4) and heart disease (2). The OC was not a common cause of MHT-amongst our sample of women of childbearing age, but a past history of hypertension in pregnancy was important. Such patients also had a longer duration of hypertension and poorer renal function at follow-up. PMID- 9349450 TI - Analysis of antibodies against components of the autonomic nervous system in diabetes mellitus. AB - Antibodies to autonomic nervous system structures have previously been detected using a complement fixation immunofluorescence test in the sera of patients with insulin-dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM). These antibodies might play a role in the aetiology of autonomic neuropathy. Sera from 45 IDDM, 40 NIDDM and 52 control subjects were tested by immunofluorescence for antibodies to human sympathetic ganglia, human adrenal medulla and rabbit vagus nerve. The use of human sympathetic ganglia was compared with rabbit tissue for the detection of sympathetic ganglia antibodies; the results for these autonomic nervous system antibodies were also compared with results using an ELISA. There was no relationship between the presence of antibodies detected by ELISA and those detected by immunofluorescence, but of 14 IDDM patients with thyroid antibodies, 12 had autonomic nervous system antibodies detected by either immunofluorescence or ELISA (p < 0.005 compared to patients without thyroid antibodies). To further characterize the autoantigen(s), immunoblotting was performed. An adrenal antigen corresponding to 74 kDa was detected in sera from three patients, only one of whom had antibodies detectable by ELISA and immunofluorescence. One IDDM serum showed specific binding to a vagus nerve antigen corresponding to 33 kDa. No specific binding to sympathetic ganglia antigen was demonstrated. Antibodies against autonomic nervous system antigens are an inconsistent feature of diabetes, and appear more associated with coincidental autoimmunity against other organs such as the thyroid. PMID- 9349451 TI - Outcome of post-transfusion hepatitis C: disease severity in blood-component recipients and their implicated donors. AB - The UK 'Look-back Program' identifies recipients of blood products from hepatitis C antibody (anti-HCV) positive donors. Of 60 such recipients tested by the Newcastle Transfusion Service, 28(46.7%) were anti-HCV-negative, 25(41.7%) were anti-HCV-positive, and seven (11.6%) had equivocal serology. We studied 29 anti HCV-positive/indeterminate recipients and eight of their implicated donors, using serial liver function tests (LFTs), liver histology when clinically indicated, HCV RNA and serotyping. Presumed resolved hepatitis C, with persistently normal LFTs and negative HCV RNA, was found in 28%, of whom 63% had indeterminate anti HCV by RIBA (1 band of 4 detected on third-generation recombinant immunoblot assay). Resolved hepatitis C was significantly more common in women (p < 0.05) and tended to be associated with younger age at transfusion. There was complete concordance in serotype between donor-recipient pairs. There was no correlation in disease severity between recipients and their implicated donors, nor between recipients from the same donor. A history of alcohol consumption above recommended 'safe' limits (median 30 units) was associated with more severe histological disease (p < 0.01). Host factors, including gender and alcohol consumption, may be important in determining the wide variability in outcome of post-transfusion hepatitis C. PMID- 9349452 TI - Elevated plasma total homocysteine and C677T mutation of the methylenetetrahydrofolate reductase gene in patients with spina bifida. AB - Total plasma homocysteine (tHcy) was significantly higher in 28 children with spina bifida (median 6.05 mumol/l) as compared with 76 controls (median 4.94 mumol/l). This difference was confined to a subgroup of patients (16/28) with one or two C677T-mutated alleles in the methylenetetrahydrofolate reductase gene. Since we found no significant difference between patients and controls in serum folate, erythrocyte folate, serum cobalamin or serum methylmalonic acid, which were within the normal range for both patients and controls, the elevated tHcy could not be attributed to vitamin deficiencies. Our findings point to an additional genetic defect involving folate-dependent enzymes in a subgroup of patients with neural-tube defects. PMID- 9349453 TI - Too much life on earth? PMID- 9349454 TI - Acidosis in severe childhood malaria. PMID- 9349455 TI - Buffer depletion and the reduction of capacity for work. PMID- 9349456 TI - Drop attacks in the elderly revisited. PMID- 9349457 TI - Seizures in cerebral malaria. PMID- 9349458 TI - Molecular virology of hepatitis C virus. PMID- 9349459 TI - Antigenic structure, evolution and immunobiology of human respiratory syncytial virus attachment (G) protein. PMID- 9349460 TI - Antigenic structure of the human respiratory syncytial virus G glycoprotein and relevance of hypermutation events for the generation of antigenic variants. AB - A set of monoclonal antibodies (MAbs) specific for the attachment (G) glycoprotein of a recently isolated strain of human respiratory syncytial virus (HRSV) is described. Antibody reactivity with a series of HRSV isolates belonging to antigenic groups A and B identified three epitope categories: (i) strain specific or variable epitopes that were present in a limited set of viruses from the same antigenic group, (ii) group-specific epitopes shared by viruses from the same antigenic group and (iii) conserved epitopes present in all HRSV isolates. Sequence analysis of escape mutants was used to map relevant antigenic sites of the G glycoprotein. Strain-specific epitopes were located preferentially in the variable C-terminal third of the G polypeptide, in agreement with previous studies of the Long strain. However, a new strain-specific epitope was mapped into another variable region, N-terminal to the cluster of cysteines in the G protein ectodomain. In contrast, the group-specific and conserved epitopes were located in the central conserved region of the G protein primary structure. These results, together with previous analysis of the Long strain, provide a detailed antigenic map of the HRSV attachment protein. Some mutants selected with group specific antibodies contain multiple A-G substitutions (hypermutations) and lack one or two of the four cysteines which are conserved in all HRSV isolates. The genetic mechanism implicated in the generation of the hypermutated viruses and its relevance for the natural history of HRSV are discussed. PMID- 9349461 TI - Variations in the neutralizing and haemagglutination-inhibiting activities of five influenza A virus-specific IgGs and their antibody fragments. AB - Neutralization and haemagglutination-inhibition (HI) of a type A influenza virus by a panel of five monoclonal IgGs, their F(ab')2s, Fabs and Fabs+ anti-mouse Fab were compared. The MAbs were specific for antigenic sites A, B and D of the haemagglutinin. Activities of the IgGs varied by up to 6-fold on a molar basis, apart from the HI activity of HC58 which was > 100-fold lower. This was not due to low functional affinity as HC58 had the second highest value (nM) as determined by an equilibrium method with whole virions. Conversion to the F(ab')2 reduced neutralization and HI by only 2- to 6-fold, indicating that the Fc region had little involvement in these processes. However, all Fabs had low neutralization and HI activity compared with their IgGs, neutralization being reduced by 86 to > 1912-fold, and HI by 13 to > 69-fold. Although decreased, their affinities remained high, in the nM range. Neutralization and HI by three of the Fabs (HC2, HC3W and HC61) were restored by the addition of anti-Fab IgG; however, HC10 Fab+anti-Fab IgG still had no detectable neutralization activity but gave HI, and HC58 Fab+anti-Fab IgG had no detectable HI activity but neutralized to the same extent as its IgG. The different properties of the antibodies are discussed in the light of their known mechanisms of action: HI by steric blocking of attachment of virus to the red cell receptor, and neutralization by the inhibition of post-attachment events (HC2, HC10 and HC61). The data demonstrate just how variable are the antiviral properties of individual IgGs. PMID- 9349462 TI - High and low efficiency neutralization epitopes on the haemagglutinin of type A influenza virus. AB - The relationship between the efficiency of the neutralization process and the affinity of five monoclonal IgG antibodies specific for the haemagglutinin of type A influenza virus has been investigated by determining their neutralization rate constants (Kneut.) and affinities (Kdissoc.). We addressed the hypothesis that if antibody affinity alone determined the efficiency of neutralization, then the Kneut.:Kdissoc. ratio would be the same for all antibodies. However, we found that the Kneut.:Kdissoc. ratio varied by up to 125-fold, suggesting that properties unique to the epitope are of major importance in determining the efficiency of neutralization. These data suggest that vaccines should preferentially stimulate antibodies to epitopes that mediate the most efficient neutralization, and that a high Kdissoc. should be an important but secondary consideration. PMID- 9349463 TI - Influenza virus NS1 protein interacts with viral transcription-replication complexes in vivo. AB - The interaction of influenza virus NS1 protein with other viral products in the infected cell was analysed by co-immunoprecipitation studies. The three subunits of the polymerase and the nucleoprotein, but not M1 protein, were co immunoprecipitated by NS1-specific serum but not when control serum was used. Such co-immunoprecipitation was not sensitive to RNase treatment of the immunoprecipitates. Co-immunoprecipitation was also obtained when the viral transcription-replication system was reconstituted in vivo by transfection of cDNAs and model vRNA template into vaccinia virus-T7-infected cells. Analysis of the RNA pulled-down in the NS1-specific precipitates indicated the presence of both vRNA and mRNA. These results are discussed in the context of the phenotype of virus temperature-sensitive mutants affected in the NS1 gene. PMID- 9349464 TI - Mx1 sensitivity: Batken virus is an orthomyxovirus closely related to Dhori virus. AB - Batken virus, isolated from mosquitoes and ticks, was tentatively classified as a member of the family Bunyaviridae. Here we show that Batken virus is inhibited by the interferon-induced Mx1 protein of mice which selectively blocks the growth of orthomyxoviruses, including Thogoto and Dhori viruses. Furthermore, we show that Batken virus multiplication is characterized by accumulation of viral proteins in the nucleus and by budding of viral particles from the cell surface. Serological cross-reactions between Batken and Dhori viruses revealed a phylogenetic relationship of these viruses, as previously also proposed by D. K. Lvov. Fragments of the Batken virus glycoprotein and nucleoprotein genes were amplified by RT-PCR. The deduced amino acid sequences were similar to the corresponding Dhori virus sequences. Therefore, Batken virus should be classified into the newly established genus Thogotovirus of the family Orthomyxoviridae. Finally, our results demonstrate that Mx1 susceptibility of orthomyxoviruses is a reliable marker in the hunt for new family members. PMID- 9349465 TI - Detection of a novel Borna disease virus-encoded 10 kDa protein in infected cells and tissues. AB - Borna disease (BD) is a transmissible, progressive polioencephalomyelitis primarily of horses and sheep. The genomes of two cell-adapted strains of Borna disease virus (BDV), the aetiological agent of BD, have been cloned and sequenced. According to the structural characterization achieved so far, BDV contains a non-segmented negative-sense 8.9 kb single-stranded RNA genome. In this paper we report the expression, purification and intracellular tracing of a novel non-glycosylated BDV-specific protein with a molecular mass of approximately 10 kDa (BDV p10 protein). The successful isolation of the corresponding mRNA from infected cells, amplification of the genetic region by RT PCR and its efficient expression as a glutathione S-transferase (GST) fusion protein demonstrated that antibodies specific for the BDV p10 protein are induced in infected animals. In addition, we have produced monospecific antisera against the GST-p10 fusion protein in rabbits. This monospecific antiserum recognized the BDV p10 protein in brain cells of naturally and experimentally infected animals as well as in persistently BDV-infected cells. Antibody-mediated affinity chromatography using the anti-p10 serum could successfully be applied to purify a ca. 10 kDa antigen from infected animal cells to such an extent that glycosylation of this component could be ruled out. PMID- 9349466 TI - Establishment of persistent hepatitis C virus infection and replication in vitro. AB - Hepatitis C virus (HCV) is a major cause of chronic viral hepatitis. Development of anti-viral strategies has been hampered by the lack of efficient cell systems to propagate HCV in vitro. To establish a long-term culture system, we tested human hepatoma (HuH7, HepG2) and porcine non-hepatoma (PK15, STE) cell lines, as well as several culture and infection conditions. As a marker for virus replication, minus-strand HCV RNA in infected cells was detected by an enhanced detection system using nested RT-PCR followed by hybridization analysis. Short term efficiency of HCV infection (10 days) was slightly increased by addition of polyethylene glycol (PEG) and/or dimethyl sulfoxide (DMSO) to culture media during inoculation of HuH7, PK15 and STE cells, but no augmentation in long-term culture was achieved, suggesting enhanced attachment of HCV to cells rather than more efficient infection. A stabilizing effect on HCV propagation was observed for 50 days in a serum-free medium with stimulation of the low-density lipoprotein (LDL) receptor expression by lovastatin. Using partially serum-free culture conditions, long-term persistence of HCV in cells and release of virions into supernatant was achieved for up to 130 days. Infectivity of released virions in supernatants after long-term culturing (day 30-80) was shown by successful infection of fresh cells. In conclusion, supplementation with PEG, DMSO and lovastatin during inoculation did not enhance virus replication substantially, but continued stimulation of LDL-receptor expression resulted in infections which persisted for over 4 months. These data support the hypothesis of an LDL-receptor mediated uptake of HCV into cells in vitro. PMID- 9349467 TI - Evolutionary analysis of the 5'-terminal region of hepatitis G virus isolated from different regions in China. AB - We have determined the nucleotide sequence of the 5'-terminal region of the hepatitis G virus (HGV) genome in 11 hepatitis patients from three cities in China. Phylogenetic analyses revealed that the Chinese isolates were genetically distinct from previously described West African isolates (type 1) and American, European and East African isolates (type 2), with a mean sequence divergence of approximately 10%. The mean divergence between isolates from Lanzhou, in the northwest of China, and those from Shanghai and Nanjing, on the east coast of China, was 5% (range 3-7%). The isolates from Shanghai and Nanjing were closely related to a common strain in Japan, while some of those from Lanzhou were closely related to a southeast Asian type 3 isolate. Thus, the Chinese isolates belong to the type 3 variant of HGV. PMID- 9349468 TI - In vivo induction of interferon-alpha in pig by non-infectious coronavirus: tissue localization and in situ phenotypic characterization of interferon-alpha producing cells. AB - A low frequency peripheral blood mononuclear cell (PBMC) subpopulation, referred to as natural interferon-producing (NIP) cells, is described as producing interferon-alpha (IFN-alpha) following contact with non-infectious viral structures, namely viral glycoproteins. These cells are characterized in vitro as non-T, non-B, MHC class II+ and CD4+ cells. In this study, NIP cells were analysed in vivo after an intravenous injection of UV-inactivated transmissible gastroenteritis virus in newborn piglets, which resulted in strong serum IFN alpha production. Splenocytes, but not PBMC, were the IFN-alpha producers in vivo. Using double immunohistochemical labelling for both IFN-alpha and leukocyte markers, we established that splenic NIP cells were not T or B cells. The majority were MHC class II+ and only a minority expressed a macrophage marker. NIP cells were localized in contact with MHC class II-expressing cells and T cells, which suggested that NIP cells might modulate the antiviral immune response. PMID- 9349469 TI - Inhibition of human immunodeficiency virus type 1 particle formation by alterations of defined amino acids within the C terminus of the capsid protein. AB - In previous studies, we demonstrated that the substitution of amino acid triplets for alanines in the carboxy-terminal portion (amino acids 341-352: ATL EEM MTA CQC) of the capsid protein domain (p24) of human immunodeficiency virus type 1 (HIV-1) partly led to an inhibitory effect on the capacity to form virus-like particles (VLPs). In these experiments, the uncleaved Pr55gag precursor protein was expressed by recombinant vaccinia viruses. We have now investigated the effects of these mutations with respect to a replication-competent HI-provirus system. Substitution of amino acids 344-346 (EEM) for alanines, which was previously shown to lead to an inhibition of VLP formation, completely blocked assembly and release of HIV. A substantial reduction of HIV synthesis was also observed in the proviral system after exchange of amino acids 347-348 [MT(A)] which, in contrast, was formerly shown to result in an increased formation of VLPs. Western blot analysis of lysates of cells transfected with these mutated proviral constructs revealed an abnormal intracellular processing pattern of the Pr55gag precursor molecules. Further analyses suggest a structural aberration of these altered polyproteins as the basis for the observed block of virus formation. PMID- 9349471 TI - Expression of naturally occurring antisense RNA inhibits human immunodeficiency virus type 1 heterologous strain replication. AB - Recently, the presence of human immunodeficiency virus type 1 (HIV-1) RNA transcripts with negative-strand polarity has been shown in tissue culture models of acute and persistently infected cells. One of these transcripts encodes a 189 amino acid open reading frame. This highly conserved antisense sequence is complementary to the structured Rev-responsive element and extends through the cleavage site of the Env protein. We tested the ability of this antisense RNA to modulate HIV-1 replication and the mRNA profile when expressed stably or transiently in several cell types. Different cell lines and PBLs were transduced by retroviral vectors producing antisense RNA and were then challenged by HIV infection. We have shown that the endogenously expressed antisense RNA containing the natural open reading frame inhibits HIV-1(IIIB) and HIV-1(NDK) replication in these cells. The level of inhibition varied according to the cells, but was significant in all cases. The production of HIV-1 (BRU, IIIB, NDK) mRNAs was also significantly decreased. HIV-2 replication was not inhibited by expression of the antisense RNA. Our results also suggest that this inhibitory effect is due to the antisense RNA and not to the protein which is encoded by this sequence. PMID- 9349470 TI - Generation of infectious virus particles by transient co-expression of human immunodeficiency virus type 1 gag mutants. AB - We have demonstrated that COS7 cells transiently co-expressing myristylation defective (Myr-) and protease-defective (PR-) human immunodeficiency virus (HIV) mutants can release infectious virions when co-transfected with an amphotropic murine leukaemia virus envelope protein expression plasmid (SV-A-MLV-env). In contrast, no infectious virions were detected when a PR-, noninfectious HIV gag mutant was co-expressed with the Myr- mutant, although the Myr- mutant could still process the immature core particles in trans. This result indicates that generation of functionally normal Gag proteins is required for virus infectivity in our complementation system. A mutant with a 56-amino-acid deletion in the N terminal region of the capsid (CA) domain could still complement the PR- mutant to generate infectious virions, suggesting that the deletion mutant could provide a functional protease for processing in the PR- mutant. This result is consistent with the concept that mutations within the N-terminal region of the CA domain have no major effects on Gag-Pol incorporation into particles. PMID- 9349472 TI - Syncytium formation induced by human immunodeficiency virus type 1 isolates correlates with affinity for CD4. AB - Different strains of human immunodeficiency virus type 1 (HIV-1) show considerable divergence in genetic content and biological properties. One property that has been closely correlated with clinical prognosis is the ability to induce syncytia formation in susceptible cells. This ability had been correlated with the V3 loop sequence of major envelope glycoprotein, gp120, but recent reports have questioned this connection. We investigated the contributions of different regions of the env gene to syncytia induction using chimeric viruses that contain part of the genome of a strain that lacks this ability (HIV-1(Ba-L)) within the genome of a virus that can form syncytia (HIV-1(HXB-2)). When tested in two cell lines susceptible to both parental viruses, as well as in primary cells, these chimeric viruses demonstrated that the ability to induce syncytia formation was determined by regions of env outside the V3 loop, which encompass residues that contribute to the binding of CD4 by gp120. Further investigation failed to show any difference in the expression of gp120 on the cell surface or cell adhesion molecules by cells infected with SI or NSI variants that would explain the observed differences in the ability to form syncytia. Assays of relative affinity for CD4 indicated that gp120 from SI variants showed a significantly higher affinity for CD4 than gp120 from NSI variants. These observations suggest that areas of the HIV-1 env gene contributing to the CD4 binding site may also contribute to the determination of syncytium-inducing (SI) and non-syncytium-inducing (NSI) phenotypes. PMID- 9349473 TI - No reactivation of attenuated immunodeficiency viruses in rhesus macaques after vaccinia virus-induced immune activation. AB - Live-attenuated simian immunodeficiency virus (SIV) protects macaques against challenge with pathogenic SIV. To evaluate the safety of such vaccines, an investigation of whether or not nef-deleted SIV could be reactivated in vivo by immune activation of the host was conducted. In addition, monkeys infected with apathogenic SIV/HIV-1 chimeric viruses, and two control monkeys that had suppressed replication of pathogenic SIV were examined. During the infection virus became undetectable or persisted at a low level of replication in all monkeys. At this time-point 11 monkeys were immune-activated by a vaccinia virus (VV) superinfection. After VV infection up to 80% of their lymphocytes showed expression of the activation markers CD25 and CD69 over 2 weeks. However, only the two non-progressing monkeys infected with pathogenic SIV showed a noticeable but transient enhancement of SIV replication and increased SIV antibody titres. By contrast, in monkeys infected with apathogenic immunodeficiency viruses no change in virus load was observed. Therefore, attenuated immunodeficiency viruses cannot be reactivated in vivo by a VV-induced immune activation. PMID- 9349474 TI - Live attenuated simian immunodeficiency virus prevents super-infection by cloned SIVmac251 in cynomolgus monkeys. AB - The ability of a live attenuated simian immunodeficiency virus (SIV) to protect against challenge with cloned SIVmac251/BK28 was evaluated in four cynomolgus macaques. The intravenous infection of the C8 variant of the SIVmac251/32H virus, carrying an in-frame 12 bp deletion in the nef gene, did not affect the CD4+ and CD8+ cell counts, and a persistent infection associated with an extremely low virus burden in peripheral blood mononuclear cells (PBMCs) was established. After 40 weeks, these monkeys were challenged intravenously with a 50 MID50 dose of SIVmac251/BK28 virus grown on macaque cells. Four naive monkeys were infected as controls. Monkeys were monitored for 62 weeks following challenge. Attempts to rescue virus from either PBMCs or bone marrow from the C8-vaccinated monkeys were unsuccessful, but in two cases virus was re-isolated from lymph node cells. The presence of the SIV provirus with the C8 variant genotype maintaining its original nef deletion was shown by differential PCR in PBMCs, lymph nodes and bone marrow. Furthermore, in contrast to the control monkeys, the vaccinated monkeys showed normal levels for CD4+ and CD8+ cells, minimal lymphoid hyperplasia and no clinical signs of infection. Our results confirm that vaccination with live attenuated virus can confer protection. This appears to be dependent on the ability of the C8 variant to establish a persistent but attenuated infection which is necessary for inducing an immune response, as suggested by the persistence of a strong immune B cell memory and by the over expression of interleukin (IL)-2, interferon-gamma and IL-15 mRNAs in PBMCs of C8 vaccinated monkeys but not in those of control monkeys. PMID- 9349475 TI - Elevation of cytokines associated with the thrombocytopenia of equine infectious anaemia. AB - Thrombocytopenia is a common finding in infection with equine infectious anaemia virus (EIAV), a lentivirus with some homology to human immunodeficiency virus (HIV). The thrombocytopenia of EIA, like that in some HIV patients, appears to have a multifactorial pathogenesis. To investigate the decreased platelet production seen in experimental EIA, the levels of three potential negative regulators of platelet production--tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta) and interferon-alpha (IFN-alpha)--were measured in serum and bone marrow of six severe combined immunodeficient (SCID) foals and ten immunocompetent EIAV-infected foals. Levels of cytokines in pre infection foal sera and bone marrow were compared with levels observed during clinical EIA. Mean serum levels of TNF-alpha and IFN-alpha were significantly higher (P < 0.05) on days -4 to 0 of thrombocytopenia than before infection. Serum TGF-beta was significantly elevated on all days except day -1 of thrombocytopenia. Bone marrow TNF-alpha levels were significantly increased in infected foals just before clinical thrombocytopenia. TGF-beta activity was not different in pre-infection and pre-thrombocytopenia bone marrows, but levels of TGF-beta protein as determined by immunohistochemical staining were significantly higher in pre-thrombocytopenia bone marrow. IFN-alpha activity in bone marrow increased just before thrombocytopenia, but the difference was not significant at P < 0.05. Serum TNF-alpha levels were 2-2.5 times higher in SCID foals on three of the days prior to thrombocytopenia than in immunocompetent foals. No significant differences were found between the levels in SCID and immunocompetent foals of serum and bone marrow TGF-beta or IFN-alpha at any of the times examined. PMID- 9349476 TI - Comparison of the complete sequence of feline spumavirus with those of the primate spumaviruses reveals a shorter gag gene. AB - The complete nucleotide sequence of the provirus of feline foamy virus (FeFV), strain F-17, was determined, and compared to the available data for human and simian spumaviruses. In addition to the usual retroviral gag, pol and env genes, two open reading frames are present between the env gene and the 3'-LTR, as in the simian spumaviruses, the first being the putative transactivator. The gag gene is predicted to encode a precursor protein of only 53 kDa compared to 70 kDa for simian spumaviruses and a doublet of 70/74 kDa for human spumavirus. The gag gene contains conserved splice acceptor and donor sites suggesting that, like human foamy virus, FeFV expresses its pol gene using a spliced mRNA. The poland envgenes showed greater sequence similarity to their counterparts in the primate spumaviruses than the gag gene and additional open reading frames. PMID- 9349477 TI - Interleukin 4 stimulates infection and temporary growth of human neonatal lymphocytes exposed in vitro to human T-lymphotropic virus type I, but fails to substitute for interleukin 2 in the immortalization of infected cultures. AB - It has been shown that interleukin 4 (IL-4) stimulates the proliferation of cells from patients affected by adult T-cell leukaemia, the haematological malignancy aetiologically associated with human T-lymphotropic virus type I (HTLV-I). In the present study, human neonatal lymphocytes were exposed to HTLV-I in vitro in the presence of IL-4. The results showed that: (i) cultures exposed to HTLV-I in the presence of either IL-4 or IL-2 bound IL-4; (ii) IL-4 did not substitute for IL-2 as a growth factor in cell lines previously infected and maintained in IL-2; (iii) cultures exposed to HTLV-I and maintained in IL-4 or IL-2 became infected; and (iv) IL-4 sustained the growth of HTLV-I-infected cultures for a maximum of 14 weeks. Moreover, HTLV-I-infected cultures grown in IL-4 showed upregulation of the IL-4 message and lower expression of HLA-DR and CD25 when compared with counterpart cultures maintained in IL-2. These results suggest that continuous growth of T-lymphocytes induced in vitro by HTLV-I infection, at least temporarily, requires signals specifically provided by IL-2 and not by IL-4. PMID- 9349478 TI - Spliced human endogenous retroviral HERV-H env transcripts in T-cell leukaemia cell lines and normal leukocytes: alternative splicing pattern of HERV-H transcripts. AB - The majority of human endogenous retroviral HERV-H elements in the human genome have large deletions in pol and lack most of env, 5-10% are more or less complete with a potentially immunosuppressive transmembrane protein-encoding env region. Spliced HERV-H env transcripts were detected in T-cell leukaemia cell lines and lymphocytes from healthy blood donors by using RT-PCR. The transcripts all contained a splice donor in the leader region downstream from the primer-binding site and a previously unreported splice acceptor in the integrase-encoding region of pol, absent in the HERV-H deletion elements. In singly spliced transcripts the leader and integrase regions were joined directly whereas in multiply spliced transcripts they were joined with an alternative exon from the protease-encoding region located between the two regions. env transcripts from three different HERV H elements were identified: one element similar to a HERV-H consensus sequence was primarily amplified from the T-cell leukaemia cell lines and two other more defective elements were amplified from normal lymphocytes. One of these elements was shown to be a reintegrated spliced transcript where the protease and integrase regions were joined, removing most of pol but leaving gag intact. Other spliced transcripts, joining the protease region and the 3'-LTR, were also amplified. The fact that HERV-H elements with an intact env splice acceptor also use the splice sites in the protease-encoding region suggests that this unusual multiple splice pattern could have a biological function in the intact HERV-H. PMID- 9349479 TI - Inhibition of Moloney murine leukaemia virus by a retroviral vector, LNL6, carrying ribozymes targeted to the 5' non-coding sequence. AB - The packaging region psi is the RNA sequence which directs the inclusion of genomic viral RNA into virions. As such it represents an apparently accessible site for ribozyme action. Ribozymes directed against the psi site of Moloney murine leukaemia virus (MoMLV) were delivered to target cells using a related retroviral vector, LNL6. LNL6 is a vector predominantly derived from MoMLV and, like all retroviruses, requires the psi region to be packaged. By exploiting the heterogeneity between nucleotide sequences of the MoMLV target and LNL6 vector in the psi packaging region, two ribozymes were designed and shown to selectively cleave the target but not the vector sequence. Clonally derived cell lines expressing ribozymes showed inhibition of virus replication. These results show the utility of catalytic RNA as specific antiviral agents. PMID- 9349481 TI - Cytokeratin patterns of expression in human epithelial cell lines correlate with transcriptional activity of the human papillomavirus type 16 upstream regulatory region. AB - A comparison of the CAT reporter activity of a plasmid which contains the 232 bp epithelial specific enhancer alone with that of plasmids which contain additional sequences from the human papillomavirus type 16 (HPV-16) upstream regulatory region (URR) revealed two markedly different patterns, in an analysis of six human epithelial cell lines. In HeLa, C33A and SiHa, the CAT reporter activities of all the constructs were comparable. In contrast, in CaSki, HK2bE6-E7 and HaCaT we detected very low levels of CAT reporter activity using the constructs with the additional HPV-16 URR sequences. The ability of HPV-16 E2 to transactivate a construct with 2 E2 binding sites also differed markedly and showed the same pattern. Cytokeratin staining revealed a correlation between cytokeratin 10 and 14 expression and the transcriptional differences observed. We also found alterations in the activity of one of the constructs on altering the growth conditions of the HaCaT cell line. PMID- 9349480 TI - Inhibition of murine leukaemia virus retrotranscription by the intracellular expression of a phage-derived anti-reverse transcriptase antibody fragment. AB - The intracellular targeting of recombinant antibodies is an experimental strategy to interfere with the function of selected molecules that is being utilized in a variety of different systems for research and medical applications. Since recombinant antibodies are increasingly being derived from phage display libraries, we have exploited phage technology to isolate, from a large combinatorial library, human antibody fragments directed against human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT). We describe in this paper the in vitro and in vivo properties of a neutralizing anti-RT antibody fragment. We demonstrate that the heavy chain domain (VH-CH1) of the phage derived antibody is able to inhibit the retroviral enzyme, in that it neutralizes the RNA-dependent DNA polymerase activity of HIV-1 RT. The VH-CH1 antibody fragment also neutralizes the activity of RT of drug-resistant HIV-1 mutants as well as that of murine retrovirus RT. To confirm the broad reactivity of the synthetic antibody fragment, we have assessed the ability of the intracellularly expressed VH-CH1 protein to interfere with murine retroviral infection. To this end, we developed an in vivo selection procedure based on the antibody-mediated resistance to a cytotoxic retrovirus and used this selection procedure to rescue, from a heterogeneous population, cells expressing the VH-CH1 antibody fragment. We finally demonstrate that the intracellular expression of the recombinant heavy chain antibody fragment leads to an efficient inhibition of viral retrotranscription by murine-based retrovirus. PMID- 9349482 TI - Human papillomavirus type 16 E7 binds to the conserved carboxy-terminal region of the TATA box binding protein and this contributes to E7 transforming activity. AB - We have previously shown that the human papillomavirus E7 proteins bind to the cellular TATA box binding protein (TBP). In this paper we show that the HPV-18 E6 and the HPV-16 E2 proteins will also bind TBP in vitro. This feature of virus proteins is conserved across many viral types and we were interested in determining whether these HPV proteins interacted with the same conserved region of the TBP molecule. A series of deletions was introduced into the TBP protein and its binding to these HPV proteins was measured. The previously well characterized interaction between p53 and TBP was used for comparison. All four proteins were found to interact with the carboxy-terminal domain of the TBP protein, although the precise residues involved and the relative strengths of association differed between the different HPV proteins. Mutational analysis of HPV-16 E7 protein identified a stretch of four amino acids responsible for the binding to TBP. This mutant E7 protein possessed wild-type levels of transcriptional activity on the adenovirus E2 promoter but exhibited reduced transforming activity in cooperation with EJ-ras. These results demonstrate that the mechanisms of interaction between diverse viral proteins and TBP are similar and that, in the case of E7, this interaction may contribute to its transforming activity. PMID- 9349483 TI - Identification of a cytotoxic T-lymphocyte epitope in the human papillomavirus type 16 E2 protein. AB - Persistent infection with oncogenic types of human papillomaviruses (HPV) is the major cause of cervical cancer precursor lesions. Cellular immune responses are considered important in the elimination of HPV infection, but the targets are not well defined. HPV E1 and E2 proteins form a replicative complex necessary for viral genome maintenance. To investigate whether epitopes in the E1 or E2 proteins can serve as targets for cytotoxic T-lymphocyte (CTL)-mediated killing, we identified peptides containing the human leukocyte antigen (HLA)-A*0201 binding motif in the deduced amino acid sequences of the HPV-16 E1 and E2 genes. Binding affinity of the peptides was measured by HLA-A*0201 up-regulation on T2 cells. Peptides with high binding-affinity were tested for their ability to elicit peptide-specific CTLs from healthy blood donors. We found one peptide from the E1 and one from the E2 protein sequence that were capable of eliciting peptide-specific CTLs. The E2-specific CTLs lysed an HPV-16-transfected cervical carcinoma cell line, but not the untransfected HPV-negative parental cell line, indicating that the identified E2 epitope can be presented to CTLs in HPV positive epithelial cells. These findings might have potentially important implications for studies of the natural history of HPV infection in relation to cervical carcinogenesis. PMID- 9349484 TI - Human cytomegalovirus late-phase maturation is blocked by stably expressed UL32 antisense mRNA in astrocytoma cells. AB - Human cytomegalovirus (HCMV) open reading frame UL32 codes for the basic phosphoprotein pp150 (ppUL32), an abundant constituent of the virion tegument. In order to study its potential role in the assembly and/or transport of progeny particles, astrocytoma cell lines (U373MG) were generated, stably expressing a 2.1 kb 5' fragment of UL32 in antisense orientation under the control of the HCMV major immediate early promoter. The steady-state level of the UL32 sense mRNA and pp150 synthesis were strongly reduced in infected antisense cell lines. Neither immediate early and early gene expression, nor viral DNA replication, was inhibited; the expression of the late gene product gB (gpUL55) was also reduced, but mainly at the level of translation. Control experiments indicated that this differential effect of UL32 antisense expression on the synthesis of viral products was specific. As a consequence of the inhibitory effect, virus yield was significantly reduced in antisense mRNA cell lines. Ultrastructural comparison of control and antisense cells revealed no difference in nucleocapsid forms in the nucleus. However, in the cytoplasm of antisense cells, DNA-containing C capsids and virions were absent and abnormal forms of non-infectious enveloped particles were observed. The data suggest the involvement of pp150 either in the transport of DNA-containing particles through the nuclear envelope or in the stabilization of capsids in the cytoplasm. Thus, UL32 antisense mRNA appears to interfere strongly with virus maturation during the late phase of the infectious cycle. PMID- 9349485 TI - Characterization of the vaccinia virus F8L protein. AB - Vaccinia virus infection dramatically affects the host actin cytoskeleton by inducing disassembly of actin stress fibres and formation of actin tails which propel the virus intra- and intercellularly. The viral factors responsible for these actin rearrangements remain unknown. Sequence analysis reveals significant homology between the vaccinia F8L ORF and the proline repeats of iActA, the protein which initiates actin tail assembly and motility in the bacterial pathogen Listeria ivanovii. We characterized the F8L gene product to examine its possible role in vaccinia rearrangements of the host actin cytoskeleton. F8L is a approximately 8 kDa protein expressed early during infection and is found throughout the cytoplasm, with no discernible association with viral or cellular structures. Furthermore, the F8L deletion strain, WR deltaF8L, forms particles and actin tails indistinguishable from WR. Our observations demonstrate that F8L is not required for vaccinia virus morphogenesis or the actin rearrangements observed during infection. PMID- 9349486 TI - Altered antigenicity of 'a' determinant variants of hepatitis B virus. AB - The 'a' determinant of hepatitis B virus (HBV) surface antigen (HBsAg) is the most important target for diagnosis and immunoprophylaxis. Several HBV variants with point mutations within the 'a' determinant have been identified among fully vaccinated children in Taiwan. We investigated the effect of each of these mutations on the antigenic nature of the S protein by cloning and expression of the mutant S antigens in Pichia pastoris. Four variants, Ser-126, His-129, Arg 129 and Arg-145, all exhibited various degrees of altered binding of HBsAg to several monoclonal antibodies. Arg-145, a well-characterized immune escape mutant, and Arg-129 had the lowest binding capacities to all monoclonal antibodies as compared with other variants. Similar to Arg-145, the Arg-129 variant could be isolated from both vaccinated children and unvaccinated adults, thus representing a naturally occurring mutant with an altered 'a' determinant. Whether these 'a' determinant variants with altered antigenicity might gradually become major circulating strains, as a consequence of the immune pressure against the wild-type HBV created by vaccination, remains to be monitored. PMID- 9349487 TI - The minute virus of mice (MVM) nonstructural protein NS1 induces nicking of MVM DNA at a unique site of the right-end telomere in both hairpin and duplex conformations in vitro. AB - The right-end telomere of replicative form (RF) DNA of the autonomous parvovirus minute virus of mice (MVM) consists of a sequence that is self-complementary except for a three nucleotide loop around the axis of symmetry and an interior bulge of three unpaired nucleotides on one strand (designated the right-end 'bubble'). This right-end inverted repeat can exist in the form of a folded-back strand (hairpin conformation) or in an extended form, base-paired to a copy strand (duplex conformation). We recently reported that the right-end telomere is processed in an A9 cell extract supplemented with the MVM nonstructural protein NS1. This processing is shown here to result from the NS1-dependent nicking of the complementary strand at a unique position 21 nt inboard of the folded-back genomic 5' end. DNA species terminating in duplex or hairpin configurations, or in a mutated structure that has lost the right-end bulge, are all cleaved in the presence of NS1, indicating that features distinguishing these structures are not prerequisites for nicking under the in vitro conditions tested. Cleavage of the hairpin structure is followed by strand-displacement synthesis, generating the right-end duplex conformation, while processing of the duplex structure leads to the release of free right-end telomeres. In the majority of molecules, displacement synthesis at the right terminus stops a few nucleotides before reaching the end of the template strand, possibly due to NS1 which is covalently bound to this end. A fraction of the right-end duplex product undergoes melting and re-folding into hairpin structures (formation of a 'rabbit-ear' structure). PMID- 9349488 TI - Efficient gene transfer into various mammalian cells, including non-hepatic cells, by baculovirus vectors. AB - A baculovirus (Autographa californica nucleopolyhedrovirus) vector containing a strong promoter, the CAG promoter, was developed to introduce foreign genes into mammalian cells. Recombinant baculoviruses carrying a reporter gene under the control of the CAG promoter were inoculated into various mammalian cell lines. High-level expression was observed not only in hepatocytes but also in other non hepatic cell lines tested. Expression of the reporter gene was detected even 14 days after infection. The infectious titre of the recovered baculoviruses decreased significantly after infection, indicating that the baculoviruses did not replicate in mammalian cells. We then compared the efficiencies of gene expression by the baculovirus vector with that of a replication-defective adenovirus vector by using the same expression unit. The same level of expression was observed in HepG2, HeLa and COS7 cells by both vectors. Efficient expression and proper processing were observed in mammalian cells infected with baculoviruses carrying genes coding for structural regions of hepatitis C virus. These results suggest that the baculovirus vector is a good tool for gene delivery into various mammalian cells in order to study the function of foreign genes. PMID- 9349489 TI - The gene encoding the capsid protein P82 of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus: sequencing, transcription and characterization by immunoblot analysis. AB - A gene encoding a capsid-associated viral structural protein has been identified and sequenced in the genome of the Choristoneura fumiferana multicapsid nucleopolyhedrovirus (CfMNPV). The gene has a 1872 nucleotide open reading frame (ORF) encoding 624 amino acids with a predicted molecular mass of 71.4 kDa. Transcription, which appeared to be initiated from a conserved GTAAG motif of baculovirus late genes, was detected at 12 h, reached a maximum at 48 h and declined at 72 h post-infection (p.i.). Part of the ORF was cloned in frame into a prokaryotic expression vector, pMAL-c2, and the fusion protein was used to generate antibodies in rabbits. It was shown, with the aid of the polyclonal antiserum, that this viral protein was detectable at 24 h p.i. in infected cells. The protein appeared as an 82 kDa band in occlusion-derived virus and as an 82 kDa band and a 72 kDa band in budded virus. Amino acid sequence comparisons revealed that this ORF had high homology with the ORF p87 (77% similarity) of Orgyia pseudotsugata (Op) MNPV and the ORF p80 (60% similarity) of Autographa californica (Ac) MNPV. Immunoblots confirmed that the CfMNPV protein had antigenic similarities to the P87 protein of OpMNPV, but not to the P80 of AcMNPV. PMID- 9349491 TI - Long-term association of tomato yellow leaf curl virus with its whitefly vector Bemisia tabaci: effect on the insect transmission capacity, longevity and fecundity. AB - The association between tomato yellow leaf curl geminivirus (TYLCV, Israeli isolate) and its insect vector, the whitefly Bemisia tabaci, was investigated. Insects that emerged during a 24 h period were caged with TYLCV-infected plants for a 48 h acquisition access period, then with egg-plants--a TYLCV non-host--for the rest of their lives. While TYLCV DNA was associated with the whiteflies during their entire adult life, the amount of capsid protein rapidly decreased and was not detectable in the insect after approximately 12 days of age. The ability of the infected whiteflies to transmit TYLCV to tomato test plants steadily decreased with age but did not disappear completely. Transmission by viruliferous insects decreased from 100% to 10-20% during their adult lifetime, compared with a decrease from 100% to 50% for non-viruliferous insects. The association of TYLCV with adult B. tabaci led to a reduction of 17-23% in their life expectancy compared with insects that had not acquired the virus, and to a 40-50% decrease in the mean number of eggs laid. These results suggest that TYLCV has some features reminiscent of an insect pathogen. PMID- 9349490 TI - Nucleotide sequence evidence for the occurrence of three distinct whitefly transmitted, Sida-infecting bipartite geminiviruses in Central America. AB - The nucleotide sequences of two Sida-infecting geminiviruses from Honduras were determined. The symptoms of both viruses are identical in Sida rhombifolia but different in Nicotiana benthamiana. An additional symptom of one virus was yellow vein clearing on infected N. benthamiana leaves. Both Sida golden mosaic viruses (SiGMV-Ho and SiGMV-Ho(yv)) have bipartite genomes (DNAs A and B). From the SiGMV Ho(yv)-infected S. rhombifolia plant two different DNA B molecules were isolated and cloned. They differ in length by 24 nucleotides [SiGMV-Ho(yv) B1 (2593 nt) and B2 (2569 nt)] and at eight nucleotide positions. Both proteins encoded by DNA B (BV1 and BC1) are affected by these substitutions. Computer analysis shows that the bipartite genomes resemble those of other whitefly-transmitted geminiviruses. From homology analyses we conclude that both viruses are closely related but distinct. Comparison with a Sida-infecting virus from Costa Rica (SiGMV-Co) showed that the two viruses from Honduras are more similar to each other than either of them are to SiGMV-Co. Exchange of SiGMV-Ho and SiGMV-Ho(yv) genomic components resulted in viable pseudorecombinant viruses. SiGMV-Ho DNA A was able to produce a viable pseudorecombinant with SiGMV-Co DNA B while the reciprocal exchange was not infectious in N. benthamiana. SiGMV-Ho(yv) DNA A and SiGMV-Co DNA B produced a viable pseudorecombinant virus whereas only pseudorecombination of SiGMV-Co DNA A with SiGMV-Ho(yv) DNA B2, and not with DNA B1, was infectious in N. benthamiana. PMID- 9349492 TI - Cap-independent leaky scanning as the mechanism of translation initiation of a plant viral genomic RNA. AB - The genome of plum pox virus contains a single open reading frame that is translated into a large polyprotein. Although the open reading frame starts at nucleotide 36 (36AUG), it is translated from the second, 147AUG, which is in a more favourable context for translation initiation. We have carried out in vitro translation and transient expression analysis in protoplasts of a nested set of substitution and deletion mutants, and the results show that no internal structure in the 5' noncoding region of plum pox virus is necessary for efficient translation initiation. On the other hand, when the cryptic 36AUG was placed in a favourable context, it turned into an efficient initiation codon in vitro. Furthermore, AUGs that were placed in a favourable context, initiating short intraleader open reading frames, repressed translation initiation from the 147AUG in vitro and in vivo. These results point to leaky scanning as the mechanism of translation initiation of plum pox virus RNA. Nevertheless, it is a peculiar leaky scanning where the initiation of translation does not require a cap structure at the 5' end. This fact is congruent with the experimentally predicted absence of a stable secondary structure at the 5' noncoding region. PMID- 9349493 TI - Intracellular ingestion and salivation by aphids may cause the acquisition and inoculation of non-persistently transmitted plant viruses. AB - Transmission of non-persistent plant viruses is related to aphid behaviour during superficial brief probes. A widely accepted hypothesis postulates that virus acquisition occurs during ingestion of plant cell contents, and inoculation during egestion or regurgitation of previously ingested sap. Although conceptually attractive, this ingestion-egestion hypothesis has not been clearly demonstrated. Furthermore, it overlooks the anatomy of the tips of the stylets (mouthparts) and, consequently, the potential role of salivation in the inoculation process. Here, we used the electrical penetration graph (EPG) technique to investigate aphid-stylet activities associated with uptake (acquisition) and release (inoculation) of two non-persistently transmitted viruses. Our results show that acquisition occurs primarily during the last sub phase (II-3) of intracellular stylet punctures, whereas inoculation is achieved during the first sub-phase (II-1). An alternative mechanism to the ingestion egestion hypothesis is proposed on the basis of our findings. PMID- 9349494 TI - Severely combined immunodeficient (SCID) mice resist infection with bovine spongiform encephalopathy. AB - Following combined intraperitoneal and intracerebral injection with bovine spongiform encephalopathy (BSE) cow brain homogenate, SCID mice show a resistance to infection in comparison with immunocompetent CB20 mice. BSE occurred in only five out of 22 challenged SCID mice, with a mean incubation period of 573 days, whereas all the CB20 mice developed the disease with a mean incubation period of 456 days. In contrast, previous studies have shown that intracerebral infection of SCID mice with a mouse-passaged scrapie strain, ME7, produces 100% incidence of disease but no replication of infectivity in spleen. The results with BSE suggest that there is little or no direct infection of the CNS in interspecies transmissions, but that processing or replication of infectivity in peripheral lymphoid tissues may facilitate subsequent spread of infection to the CNS. PMID- 9349495 TI - Effect of Bcr sequences on the cellular function of the Bcr-Abl oncoprotein. AB - In Philadelphia chromosome (Ph1)-positive human leukemia, the c-Abl tyrosine kinase is activated by fusion to sequences encoded by the breakpoint cluster region (bcr) gene. Two major types of Bcr-Abl fusion proteins have been found in human leukemia. Fusion of the N-terminal 426 amino acids of Bcr generates p190(Bcr-Abl) which is mostly found in acute lymphocytic leukemia (ALL), whereas fusion of the N-terminal 902 or 927 amino acids of Bcr generates p210(Bcr-Abl) mostly found with chronic myelogenous leukemia (CML). Previous studies have demonstrated that both the Bcr and the Abl functional domains contribute to the oncogenic activity of Bcr-Abl proteins. Present in both p190 and p210 is the N terminal coiled-coil of Bcr (aa 1-63), which is shown here to be functionally replaceable with the leucine zipper of the yeast transcription factor GCN4. The ZIP-Bcr-Abl protein transforms Rat-1/myc cells, is autophosphorylated on tyrosine and localized predominantly to actin filaments. Thus, formation of homo-oligomers through either Bcr or GCN4 coiled-coil can activate the tyrosine kinase and F actin binding functions of Abl. We also found that a Bcr-Abl fusion containing only Bcr amino acids (1-191) can efficiently transform Rat-1/myc cells. Fusion of additional Bcr sequences (aa 192-923) did not affect the transformation of Rat 1/myc cells but progressively reduced the disruptive effect on the actin cytoskeleton. In particular, the Dbl homology domain present in p210(Bcr-Abl) but not in p190(Bcr-Abl) contributes to the stabilization of actin fibers. The modulatory effect of Bcr sequences on actin structure may underlie the apparent pathogenic variations between the different Bcr-Abl fusion proteins. PMID- 9349496 TI - Rapamycin and p53 act on different pathways to induce G1 arrest in mammalian cells. AB - Certain growth regulatory kinases contain a common domain related to the phospho inositol 3 (PI-3) kinase catalytic site. These include the ATM gene product, DNA PKcs, and the target of rapamycin (TOR in yeast; and FRAP in mammalian cells). Rapamycin inhibits growth factor signalling and induces G1 arrest in many cell types. Some growth regulatory PI-3 kinases appear functionally linked to p53 and we have explored potential links between cellular effects induced by rapamycin and p53. In p53 null cells rapamycin inhibited cell cycling but did not induce G1 arrest. In cells which showed selective G1 arrest in response to rapamycin, rapamycin had no effect on basal levels of p53 protein. Similarly p21(WAF1) protein was not induced by rapamycin. The kinetics of the cellular p53/p21(WAF1) response to ionising radiation was unaffected by rapamycin; and the ability of growth factor to protect against p53-mediated apoptosis in response to DNA damage was also unaffected by rapamycin. The ATM gene is mutated in the cancer susceptibility syndrome ataxia telangiectasia (AT) but such mutant cells showed a similar sensitivity to rapamycin compared to their normal counterparts. RKO cell lines of common genetic background, but with different levels of functional p53 protein, also responded similarly to rapamycin. Thus, although rapamycin and p53 are each able to induce G1 arrest, they appear to act through independent growth regulatory pathways. PMID- 9349497 TI - Oncogenic transformation potentiates apoptosis, S-phase arrest and stress-kinase activation by etoposide. AB - The exact mechanisms for the selective toxicity of chemotherapeutic drugs against tumor cells are not fully understood. We designed a series of experiments to test the possibility that the positive proliferative signal initiated by oncogenes might change the sensitivity for apoptosis induction by the anticancer drug etoposide (VP16), an inhibitor of topoisomerase II (Topo II). Treatment with VP16 induced significantly increased apoptosis in NIH3T3 cells transformed by oncogenic src, ras or raf, compared with the normal 3T3 cells. Apopototic changes involved nuclear DNA fragmentation, morphological alterations and decreased viability. Furthermore it was shown that stress-activated protein kinase (SAPK) was activated much more strongly in all three transformed lines compared to untransformed cells by VP16 treatment, while slight activation of extracellular signal-regulated kinase (ERK1) was observed in all four cell lines. In addition, the transformed cells displayed arrest in mid-S-phase following the treatment, whereas NIH3T3 cells were primarily arrested in late S and G2/M phase. Finally, the cyclin-dependent kinase inhibitor p21 WAF1 was induced in all four cell lines, although induction of p53 was not detected in any of these cell lines. Taken together our results demonstrated that oncogenic transformation can sensitize the cells to apoptosis induction, stress kinase activation and cell cycle arrest in response to VP16 treatment. These results may have important implications for understanding the selective toxicity of anti-cancer drugs in tumor cells. PMID- 9349498 TI - Frequent deletions of FHIT and FRA3B in Barrett's metaplasia and esophageal adenocarcinomas. AB - The FHIT gene, which spans the common fragile site FRA3B, has been shown to produce aberrant transcripts in a variety of tumor types. Homozygous deletions within the FHIT locus have been detected only in tumor-derived cell lines and LOH has been described in numerous primary tumors. Based on these findings and its location on 3p, FHIT has been proposed as a tumor suppressor gene. To further study the relationship of FRA3B to the findings regarding the FHIT gene and to determine the extent of FHIT mRNA alterations in early stages of tumor development, the status of the FHIT gene was evaluated in the premalignant condition of Barrett's esophagus and associated esophageal adenocarcinomas. FHIT expression was investigated by RT-PCR in normal esophageal, Barrett's metaplasia and adenocarcinoma tissues from 15 patients. Alterations of FHIT transcripts were observed in 12/14 (86%) of Barrett's metaplasia and in 14/15 (93%) of the adenocarcinomas from the same patients. Characterization of the altered transcripts revealed FHIT mRNA lacking one or more exons, with deletion of exons 5-7 being most frequent. Analysis of genomic DNA from 20 patients showed homozygous deletions involving exon 5 of FHIT in 4/20 (20%) esophageal adenocarcinomas, and 7/20 (35%) tumors demonstrated hemizygous loss. Genomic deletions also involved the BE758-6 locus, indicating that a large region is deleted. Fluorescence in situ hybridization (FISH) analysis demonstrated that this region of deletion is localized within FRA3B. Our results extend the range of tumor types in which altered FHIT transcripts have been demonstrated and show that these alterations can be seen in the premalignant stage of esophageal tumor development. These results indicate that the fragility and recombination-prone nature of FRA3B is related to tumor-specific chromosomal instability affecting the FHIT gene in esophageal adenocarcinoma development. PMID- 9349499 TI - Activation of the c-fos SRE through SAP-1a. AB - TCFs, which are members of the Ets family of transcription factors, are recruited to the Serum Response Element (SRE) in the c-fos promoter by SRF. These Ets proteins, which are substrates for the MAP kinases, are direct targets of the Ras/MAP kinase signal transduction pathway. In this paper, we demonstrate that one of the TCFs, SAP-1a, displays a significant level of autonomous binding to the SRE Ets box. In contrast to previous observations, deletion of the SRF binding domain did not modulate the autonomous binding of SAP-1a. Also, the autonomous binding was not modulated by the phosphorylation of SAP-1a by MAP kinases. The autonomous binding was also detected in live cells: transfected SAP 1a was able to restore the response of a CArG-less SRE in PC12 cells. The response occurred in the absence of SRF recruitment since a mutant of SAP-1a in which the B-box, a domain required for interaction with SRF, had been deleted was still able to transactivate the CArG-less SRE. The transactivation was repressed by a Ras transdominant negative mutant, indicating the involvement of the Ras/MAP kinase pathway. Taken together, these data demonstrate that SAP-1a is capable of binding to the c-fos SRE in the absence of SRF. PMID- 9349500 TI - Characterization of changes in gene expression associated with malignant transformation by the NF-kappaB family member, v-Rel. AB - In this study, alterations in gene expression patterns have been examined in v Rel-transformed avian bone marrow cells. Using a conditional v-Rel estrogen receptor chimera (v-RelER) which transforms cells in an estrogen-dependent manner, we constructed subtraction cDNA libraries from v-RelER-transformed bone marrow cells. Several different sequences were identified whose expression was altered upon hormone activation of v-RelER. These include two genes related to the MIP-1 chemokine family (mip-1beta and a tca3 homologue), a cell surface antigen sca-2 and the transcription factor nfkb1. The expression of each gene was assayed in a number of wild-type and mutant v-Rel-expressing fibroblast and hematopoietic cells. All v-Rel-transformed hematopoietic cells tested express high levels of nfkb1 and sca-2. In fibroblasts, wild-type v-Rel induced expression of mip-1beta and nfkb1, while nontransforming mutants of v-Rel failed to do so, suggesting a role for these two genes in v-Rel mediated transformation. Finally, these genes are expressed at high levels in cells overexpressing wild type and truncated forms of c-Rel, implying that v-Rel transforms, in part, by induction of c-Rel target genes. PMID- 9349501 TI - Chimeric MLL products with a Ras binding cytoplasmic protein AF6 involved in t(6;11) (q27;q23) leukemia localize in the nucleus. AB - In infantile leukemias and therapy-related leukemias, the MLL gene is frequently found to be disrupted and fused to various translocation partner genes, such as AF4/FEL, LTG9/AF9 and LTG19/ENL as a result of 11q23 translocations. We previously showed that the N-terminal portion common to various chimeric MLL products, as well as to MLL-LTG9 and MLL-LTG19, localizes in the nuclei, and therefore suggested that it might play an important role in leukemogenesis. In the present study, MLL-AF6 chimeric products found in the t(6;11)(q27;q23) translocation were analysed since AF6, a Ras-binding protein, exhibits a different subcellular localization from that of LTG9/AF9 and LTG19/ENL. Immunofluorescence staining data and cell fractionation analyses demonstrated that MLL-AF6 chimeric products localize in the nuclei despite the fact that AF6 itself localizes in the cytoplasm, confirming the importance of the nuclear localization of chimeric MLL products. The region in the N-terminal portion of MLL responsible for this nuclear localization was examined and found to be a region containing AT-hook motifs. PMID- 9349502 TI - An activated mutant of R-Ras inhibits cell death caused by cytokine deprivation in BaF3 cells in the presence of IGF-I. AB - R-Ras belongs to a family of low molecular weight GTP-binding proteins and exhibits 55% amino acid identity to H-Ras. It has been demonstrated that H-Ras inhibits cell death caused by interleukin-3 (IL-3) withdrawal in BaF3 cells (Kinoshita et al. (1995b); Terada et al. (1995)). In the present study, we examined whether R-Ras also rescues BaF3 cells from the factor-deprived cell death. To do this, several BaF3 transfectants were established, in which expression of wild-type as well as mutant R-Ras was regulated by an inducible promoter. Using these transfectants, we found that expression of an activated R Ras mutant, R-Ras (Q87L), suppressed the death of IL-3-deprived BaF3 cells. On the other hand, expression of the wild-type and the dominant-negative mutant of R Ras showed no inhibitory effect on cell death, indicating that R-Ras x GTP abrogated cell death caused by deprivation of IL-3. Furthermore, it was found that IGF-I in serum was required for the anti-apoptotic activity of R-Ras. Suppression of cell death by R-Ras(Q87L) was inhibited by wortmannin, LY294002 (phosphatidylinositol 3-kinase (PI3K) inhibitors), or PD98059 (inhibitor for MEK, a specific activator of mitogen-activated protein kinase (MAPK)). In addition, we have shown that, in HEK293 cells, R-Ras and IGF-I could activate MAPK synergistically. Also, PI3K activity was co-immunoprecipitated with an activated mutant of R-Ras. These results suggest that R-Ras in collaboration with IGF-I suppressed apoptotic cell death of BaF3 caused by IL-3 deprivation, presumably by modulating the activitites of MAPK and PI3K. PMID- 9349503 TI - Genetic alterations of chromosome band 9p21-22 in head and neck cancer are not restricted to p16INK4a. AB - Although genetic alterations of chromosome band 9p21-22 occur frequently in head and neck squamous cell carcinoma (HNSCC) cell lines, alterations of the cyclin dependent kinase inhibitor p16INK4a located in this region are less common in corresponding primary tumors. To further investigate genetic alterations at 9p21 22 and p16INK4a in primary HNSCC, a paired set of 21 tumors and blood specimens that were shown previously to exhibit allelic loss at 3p and elsewhere, were tested for LOH at 9p21-22 using eight different highly polymorphic marker. Sixteen of the samples (81%) exhibited LOH for at least one marker. Frequent LOH was found surrounding p16INK4a and at three additional non-contiguous regions of 9p21-22. No homozygous deletions were identified. SSCP screening and direct sequence analysis led to the identification of mutations the p16INK4a gene in two tumors. p16INK4a was not hypermethylated in any of the samples studied. Furthermore, there was no correlation between LOH at 9p21-22 and the RB1 tumor suppressor gene. These findings indicate that in the set of tumors that we tested, LOH at 9p21-22 is common in primary HNSCC but that genetic alterations of p16INK4a located in this region are unusual. Additional tumor suppressor genes at 9p21-22 may therefore be involved in the pathogenesis of this tumor. PMID- 9349504 TI - Involvement of p21(WAF1/Cip1), CDK4 and Rb in activin A mediated signaling leading to hepatoma cell growth inhibition. AB - Cytokines are growth inhibitory in a target cell specific manner. The signaling pathways that characterize each cell type play a crucial role in determining the responsiveness to cytokine triggering. Activin A has been shown to suppress the growth of primary hepatocytes. Similarly, the human HepG2 hepatoma cell line was growth arrested by activin A as judged by lack of cell proliferation and suppression of DNA synthesis. In HepG2 cells activin A further induced accumulation of retinoblastoma protein in the hypophosphorylated form known to prevent entrance into S phase. This finding implies the involvement of cyclin dependent kinases and CDK inhibitors. Examination of HepG2 cells following addition of activin A revealed reduced expression of CDK4 and conversely, an increase in the CKI p21(WAF1/Cip1). This accumulation of p21(WAF1/Cip1) protein was partly due to increased transcriptional activity. Functional inactivation of p53, using a miniprotein that oligomerizes with p53 and abrogates DNA binding, abolished the ability of activin A to induce transcriptional activation from the p21(WAF1/Cip1) promoter. Thus, activin A, like transforming growth factor beta, seems to suppress cell growth through the downstream target Rb. However, each of these cytokines seem to operate through a distinct pathway. PMID- 9349505 TI - Allelic profiles of mononucleotide repeat microsatellites in control individuals and in colorectal tumors with and without replication errors. AB - We have recently shown that analysis of BAT-26, was sufficient to establish the Replication Error status of colorectal tumors and cell lines without the need for matching normal DNA. BAT-26, a poly(A) tract in the 5th intron of the hMSH2 gene, does not present significant size variation either between the alleles of one individual or between alleles of different individuals. In colorectal tumors without defects in the replication error system (RER- phenotype), BAT-26 is also quasi-monomorphic. On the contrary, in RER+ colorectal tumors, BAT-26 shows unstable shortened alleles. In order to see whether this behaviour was specific for BAT-26, or was a more general phenomenon for mononucleotide repeat microsatellites, we analysed eight other mononucleotide repeats. In control individuals (72 samples) and in RER- colorectal tumors and cell lines (55 samples), these microsatellites were polymorphic, dimorphic, quasi-monomorphic or monomorphic, indicating that the quasi-monomorphic nature of BAT-26 was not a general rule. All of them showed a tendency to be shorter in RER+ colorectal tumors and cell lines (19 samples), but only quasi-monomorphic and monomorphic mononucleotide repeats could be used to determine the RER status of tumors without matching normal DNA, although with a lower efficiency than BAT-26 due to either a smaller range of shortening in RER+ tumors or to a larger number of false negative cases. PMID- 9349506 TI - Evidence of genetic progression in human gastric carcinomas with microsatellite instability. AB - Mutator phenotype tumors provide unique opportunities to unravel malignant progression because of various gene alterations acquired during clonal tumor evolution. Gastric carcinomas, which have been known to show frequent genetic instability, would be composed of initial gene alterations shared by most tumor areas and subsequent alterations restricted to particular tumor sites. To analyse the timing of genetic events, we examined separate sites of tumor tissue obtained from a given gastric carcinoma patient with microsatellite instability (MSI). Our study included 95 normal/tumor area pairs from 25 patients. Six of the 25 patients (24%) demonstrated various levels of MSI ranging from 7% (two of 30) to 97% (28 of 29) of markers tested in multiple tumor sites. Of the six patients, five manifested frameshift mutations in a tract of ten deoxyadenosines within transforming growth factor beta receptor type II and four demonstrated frameshift mutations in a tract of eight deoxyguanosines within BAX. These mutations were common to all tumor sites regardless of the various level of MSI phenotype, indicating initial events. Two of the six patients exhibited frameshift mutations in mononucleotide repeats of mismatch repair genes, hMSH3 and hMSH6, and the insulin-like growth factor II receptor in restricted tumor areas, indicating additional alterations. Insulin-like growth factor II receptor mutations appear to be caused by hMSH3 and hMSH6 mutations because the former mutations were confined to tumor portions with the latter two mismatch repair lesions. These results provide genetic progression evidence for gastric carcinomas of the mutator pathway. In this pathway, mismatch repair insufficiency initially targets mononucleotide tracts of transforming growth factor beta receptor type II and BAX. During tumorigenesis, primary mismatch repair failure may give rise to the secondary mismatch repair lesions, frameshift mutations of hMSH3 and hMSH6, which result in another tumorigenic mutation in the insulin-like growth factor II receptor. PMID- 9349507 TI - p53 is phosphorylated in vitro and in vivo by the delta and epsilon isoforms of casein kinase 1 and enhances the level of casein kinase 1 delta in response to topoisomerase-directed drugs. AB - The p53 tumour suppressor protein plays a key role in the integration of stress signals. Multi-site phosphorylation of p53 may play an integral part in the transmission of these signals and is catalysed by many different protein kinases including an unidentified p53-N-terminus-targeted protein kinase (p53NK) which phosphorylates a group of sites at the N-terminus of the protein. In this paper, we present evidence that the delta and epsilon isoforms of casein kinase 1 (CK1delta and CK1epsilon) show identical features to p53NK and can phosphorylate p53 both in vitro and in vivo. Recombinant, purified glutathione S-transferase (GST)-CK1delta and GST-CK1epsilon fusion proteins each phosphorylate p53 in vitro at serines 4, 6 and 9, the sites recognised by p53NK. Furthermore, p53NK (i) co purifies with CK1delta/epsilon, (ii) shares identical kinetic properties to CK1delta/epsilon, and (iii) is inhibited by a CK1delta/epsilon-specific inhibitor (IC261). In addition, CK1delta is also present in purified preparations of p53NK as judged by immunoanalysis using a CK1delta-specific monoclonal antibody. Treatment of murine SV3T3 cells with IC261 specifically blocked phosphorylation in vivo of the CK1delta/epsilon phosphorylation sites in p53, indicating that p53 interacts physiologically with CK1delta and/or CK1epsilon. Similarly, over expression of a green fluorescent protein (GFP)-CK1delta fusion protein led to hyper-phosphorylation of p53 at its N-terminus. Treatment of MethAp53ts cells with the topoisomerase-directed drugs etoposide or camptothecin led to increases in both CK1delta-mRNA and -protein levels in a manner dependent on the integrity of p53. These data suggest that p53 is phosphorylated by CK1delta and CK1epsilon and additionally that there may be a regulatory feedback loop involving p53 and CK1delta. PMID- 9349508 TI - p53 mutations implicate sunlight in post-transplant skin cancer irrespective of human papillomavirus status. AB - Mutations in p53 were detected in 11/23 (48%) of non melanoma skin cancers in renal allograft recipients and in 5/8 (63%) of sporadic tumours from immune competent patients. 9/12 (75%) of mutations in transplant patients and all 5 mutations in non transplant tumours were consistent with damage caused by ultraviolet (u.v.) irradiation. DNA sequences, predominantly of the epidermodysplasia verruciformis (EV) subgroup, were detected in 9/23 (39%) of transplant tumours and in 2/8 (25%) of eight non-transplant tumours. There was no relationship between HPV status and p53 mutation, HPV DNA being present in 5/16 (31%) of tumours with p53 mutation and 6/15 (40%) of tumours lacking p53 mutation. These data are consistent with an important role for sunlight in the development of post-transplant skin cancer, and with limited functional data suggesting that E6 proteins of the cutaneous and EV-related papillomaviruses do not target p53 for ubiquitin-mediated degradation. PMID- 9349509 TI - Identification of an Efs isoform that lacks the SH3 domain and chromosomal mapping of human Efs. AB - Efs was originally found by expression cloning of a mouse embryo cDNA library through its Fyn-SH3 binding capacity (Ishino et al., Oncogene 11, 2331-2338, 1995). Efs has characteristic regions important in intracellular signal transduction; these are an SH3 domain, a cluster of putative ligands for SH2 domains and proline-rich sequences with SH3-binding consensus. In this paper, we report cDNA cloning of human Efs and a variant of it from a hippocampal cDNA library. The human Efs gene was mapped to chromosome 14q11.2-q12 by fluorescence in situ hybridization. We identified two forms of human Efs, designated hEfs1 and hEfs2. hEfs1 represents the human counterpart of original mouse embryo Efs (mEfs1). hEfs2, the newly identified form, is identical to hEfs1, except for its lack of the SH3 domain. hEfs1 and mEfs1 are 80% identical in their amino acid sequences and 100% identical within the SH3 domain. Reverse transcription polymerase chain reaction analysis of adult mouse tissue RNA indicated expression of Efs2 and of Efs1 in various tissues. Evidence suggesting the presence of the Efs2 protein in human tissue was obtained by immunoprecipitation followed by immunoblotting with two different anti-Efs antibodies. Possible functions of Efs2 are discussed. PMID- 9349510 TI - Induction of telomerase activity by UV-irradiation in Chinese hamster cells. AB - Telomerase is a ribonucleoprotein whose activity has been detected in germline cells and in neoplastic and immortal cells. Telomerase compensates the telomere loss arising by the end replication problem by synthesizing telomeric repeats at the 3' end of the eukaryotic chromosomes. Telomerase is reactivated during cancer progression in human and mice. In order to determine whether the telomerase activity can be upregulated in vitro in response to DNA damaging agents, we examined the telomerase activity in five Chinese hamster cell lines following exposure to 5 J/m2 or 40 J/m2 UV-C radiation. All the cell lines tested showed an increase in telomerase activity in the PCR-based telomeric repeat amplification protocol (TRAP) in a dose dependent manner. This increase in telomerase activity correlated well with the number of cells being in the S and G2/M phase after UV exposure. However, in unirradiated control cells, similar levels of telomerase activity were observed in different phases of the cell cycle. Furthermore, telomeric signals were clustered in one or more parts of the disintegrating nuclear particles of the apoptotic cell as detected by fluorescence in situ hybridization (FISH). This is the first study to demonstrate the induction of telomerase activity following exposure to DNA-damaging agents like UV radiation in Chinese hamster cells in vitro. PMID- 9349511 TI - Despite potential flaws, the false-negative proportion remains the best practical measure of the accuracy of cervical cytology screening. PMID- 9349512 TI - Analysis of error in calculating the false-negative rate in the interpretation of cervicovaginal smears: the need to review abnormal cases. AB - BACKGROUND: Determining the false-negative rate (FNR) of cervicovaginal smear interpretation is a necessary step for any quality assessment and improvement program. All tests estimate the FNR, but the accuracy of these estimates varies from test to test. Two methods for determining the FNR have been proposed, specifically "seeding" of the initial screening population with smears from patients with a known diagnosis and rescreening a random sample of negative smears. However, the accuracy of neither method is known. METHODS: A review of the literature, an analysis of the sources of error, and an estimate of their magnitude was performed for each method. RESULTS: Seeding has a large sampling error, and more important, the FNR that this test measures does not reflect the FNR of the laboratory as a whole. Random rescreening underestimates the FNR of primary screening by the FNR of rescreening. Currently, the FNR of rescreening is not known, not measured, and may be high. Nevertheless, the FNR of rescreening and the false-positive rate (FPR) of initial screening both can be measured by rescreening abnormal cases. Knowledge of both the FNR and the FPR of initial screening allows the efficiency of cervicovaginal smear interpretation to be measured, which may be a better measure of overall accuracy than the FNR alone. CONCLUSIONS: Random, blinded rescreening of normal and abnormal smears can more accurately measure the FNR of screening than rescreening of normal smears alone. PMID- 9349513 TI - A more accurate measure of the false-negative rate of Papanicolaou smear screening is obtained by determining the false-negative rate of the rescreening process. AB - BACKGROUND: The false-negative rate (FNR), or fraction, of Papanicolaou (Pap) smear screening has been proposed as a useful quality assessment measure. The FNR should account for the FNR of the rescreening process itself. The authors measured the FNR of the rescreening process by rescreening a set of abnormal smears. METHODS: A randomly selected group of negative (150) and abnormal (91) smears were rescreened in a blinded fashion. A diagnosis of atypical squamous cells of undetermined significance (ASCUS) or worse was used as a positive (abnormal) result. All discrepancies were confirmed by consensus review. The true FNR of screening Pap smears was calculated as: true FNR = calculated FNR/(1-FNR of rescreening). RESULTS: When rescreened, 17 originally negative cases were interpreted as ASCUS and 5 as unsatisfactory. Twenty-three originally abnormal cases (22 ASCUS and 1 low grade squamous intraepithelial lesion) were interpreted as negative. After consensus review, only 1 of the originally negative cases was believed to be ASCUS and 1 unsatisfactory; 18 of the 23 originally abnormal cases were believed to be rescreening errors and 5 of the 23 originally abnormal cases were believed to be false-positives. The FNR of Pap smear screening as traditionally calculated was 6.1%, which was slightly less than the laboratory's usual FNR. The FNR of review screening was 20.9%. The true FNR of Pap smear screening was 7.8% and the false-positive rate was 0.6%. CONCLUSIONS: The FNR of rescreening is not insubstantial. It can and should be measured by rescreening abnormal smears, and when taken into account yields a more accurate measure of the FNR of Pap smear screening. PMID- 9349514 TI - Efficacy of fine-needle capillary biopsy in the assessment of patients with superficial lymphadenopathy. AB - BACKGROUND: Fine-needle aspiration biopsy (FNAB) is an important tool for the cytologic assessment of patients with lymphadenopathy. The nonaspiration method (without the use of a syringe or a handle), or fine-needle capillary biopsy (FNCB), is becoming popular for its ease of learning and use. The authors compared the two techniques of fine-needle biopsy (with and without aspiration) for identifying the cause of superficial lymphadenopathy. METHODS: Over a 2-year period 50 cytologic examinations were conducted in patients with superficial lymphadenopathy. Both procedures were performed at the same site; the order in which they were performed was determined randomly. Slides were fixed in a similar manner and examined by a cytotechnologist and pathologists blind to the procedure. Scores were tabulated and compared and diagnosis was confirmed by histologic examination. RESULTS: The best and the average score for the yield of cellular material was higher with FNCB (P < 0.004). Cell preservation also was superior with FNCB (P = 0.00066). However, the failure rate was lower with FNAB (P = 0.7662). CONCLUSIONS: FNCB of superficial lymph nodes yields adequate cellular material of superior quality, which may be advantageous in certain situations. However, the diagnostic yield does not appear to be significantly increased using this technique. PMID- 9349515 TI - Aspiration and imprint cytopathology of salivary duct carcinoma. AB - BACKGROUND: Salivary duct carcinoma (SDC) is a highly malignant primary epithelial neoplasm of the salivary glands. The histology of this lesion resembles that of cribriform and comedo-type intraductal carcinoma of the breast, yet the cytopathology of salivary duct carcinoma has been described in only a few instances in the past. METHODS: The authors report two cases of SDC diagnosed by fine-needle aspiration biopsy and one case imprinted after resection. The cytologic features that may permit diagnosis of this entity are described. RESULTS: Cytologic features include variably cellular smears containing loose clusters of cribriform and glandlike aggregates; monomorphic polygonal cells with round-to-oval nuclei; abundant, finely granular cytoplasm; indistinct nucleoli; and variable necrosis. CONCLUSIONS: The cytologic findings for SDC are relatively unique and may allow for its specific diagnosis on aspiration cytopathology. PMID- 9349516 TI - Comparison between Ki-67 index and S-phase fraction on fine-needle aspiration samples from breast carcinoma. AB - BACKGROUND: Fine-needle aspiration (FNA) biopsy has been used increasingly in the diagnosis and biologic characterization of breast carcinomas in patients who receive preoperative chemotherapy. Because proliferative activity of breast carcinoma has been shown to be of prognostic significance, the authors compared immunocytochemical Ki-67 growth fraction and flow cytometric S-phase fraction (SPF), both evaluated on FNA samples. METHODS: The proliferative activity of 134 FNA samples from primary breast carcinoma patients was studied using both immunocytochemistry with the monoclonal antibody Ki-67 and SPF determined by DNA flow cytometry. RESULTS: Ki-67 and SPF were evaluable in 114 and 107 cases, respectively, and both were evaluable in 95 cases. Of the 134 FNA samples studied, 37% were diploid and 63% were aneuploid. The distribution of both Ki-67 and SPF was different in diploid and aneuploid tumors. The median Ki-67 value as well as the median SPF were significantly higher in aneuploid versus diploid tumors (P < 0.001). Median Ki-67 and SPF values were used to discriminate between low versus high proliferating tumors. The overall concordance between Ki-67 and SPF was 75% (P < 0.001). A good correlation was found between Ki-67 and SPF (correlation coefficient = 0.72; P < 0.001). CONCLUSIONS: The results of the current study suggest that Ki-67 growth fraction and SPF determined by FNA may be used as measurements of the proliferative activity of breast carcinoma. The authors recommend these determinations be used as preoperative procedures in patients with a cytologic diagnosis of breast carcinoma who are candidates for neoadjuvant chemotherapy and/or endocrine therapy. PMID- 9349517 TI - The cell adhesion molecule, E-cadherin, distinguishes mesothelial cells from carcinoma cells in fluids. AB - BACKGROUND: The distinction between benign reactive mesothelial cells and well differentiated carcinoma can be difficult in pleural, peritoneal, and especially pericardial fluids. E-cadherin is an adhesion protein that is specifically expressed in cells of epithelial lineage. In this study, anti-E-cadherin antibodies were used to identify and distinguish carcinoma cells from reactive mesothelial cells. METHODS: Pleural, peritoneal, and pericardial fluids were prepared using the Cytyc Thin Prep processor. The specimens were comprised of a mix of 45 cases that were diagnosed as carcinoma, suspicious, or reactive by Papanicolaou staining of routine material seen by the authors' service. Routine immunologic techniques were used with a commercially available E-cadherin antibody. RESULTS: In most cases of carcinoma, tumor cells showed a strong positive membranous reaction product (32 of 37). This included four cases that were not cytomorphologically diagnosed as malignant, but subsequently proved to be malignant. E-cadherin staining was not observed in five tumors, two of which were not expected to express this protein. One benign case showed cells staining for E-cadherin, although the cells were not malignant by morphologic criteria. Because this case was a surgical pelvic washing, these cells more likely were epithelial contaminants than true false-positives. CONCLUSIONS: The epithelial specific cell-cell adhesion marker E-cadherin reliably distinguishes reactive mesothelial cells from carcinoma and is a useful adjunctive test to distinguish benign reactive mesothelial cells from well differentiated carcinoma cells in fluid specimens. PMID- 9349518 TI - Detection of hyperdiploid malignant cells in body cavity effusions by fluoresence in situ hybridization on ThinPrep slides. AB - BACKGROUND: Benign body cavity effusions sometimes cannot be distinguished from malignant ones by conventional cytology. The authors performed fluorescence in situ hybridization (FISH) on ThinPrep slides using chromosome specific probes to see if hyperdiploid malignant cells could be detected in 20 body cavity effusions. The results were then compared with those of conventional cytology. METHODS: A total of 20 body cavity effusions from 19 patients were studied using conventional cytology and FISH. Probes specific for chromosomes 3, 8, 10, and 12 were used to detect hyperdiploidy on ThinPrep slides (Cytyc Corporation, Boxborough, MA). RESULTS: A total of 13 patients had malignant conditions (either prior history of malignancy or the presence of malignancy anywhere in the body). Conventional cytology and FISH were both positive in 5 of these patients (6 samples) and negative in 2 patients. The results for one sample were inconclusive by both methods. There were 5 discrepant cytology-FISH results in patients with malignant conditions. One sample was positive by FISH and negative by cytology, one was positive by FISH and "atypical" by cytology, and three were inconclusive by FISH and negative by cytology. FISH results were either negative (in 4 samples) or inconclusive (in 2 samples) in the 6 patients with benign conditions. CONCLUSIONS: FISH can detect hyperdiploid malignant cells in body cavity effusions and is especially useful when the major cell population consists of malignant cells that cannot be differentiated from mesothelial or "atypical" cells. It is less useful in detecting a small population of malignant cells hidden in an inflammatory or reactive cell background. More studies are needed to establish diagnostic criteria further and to assess the clinical usefulness of this procedure. PMID- 9349519 TI - Detection of chromosomal aberrations in cytologic brush specimens from head and neck squamous cell carcinoma. AB - BACKGROUND: Detection of genetic changes in the mucosa of the upper aerodigestive tract may provide a target for the screening of cytologic specimens to identify premalignant transformation in this region. In this pilot study, the feasibility of the fluorescence in situ hybridization (FISH) technique to detect genetically aberrant cells in brush specimens was evaluated. METHODS: Brush specimens taken from the tumors of 20 patients with head and neck squamous cell carcinoma (HNSCC) and from the normal mucosa of 8 control patients were analyzed by FISH using DNA probes for the chromosomes 1 and 7. The FISH results were compared with DNA flow cytometry and FISH results of the solid tumor specimens. RESULTS: The results of this study showed that 15 of the 20 tumor brush specimens contained numeric chromosomal aberrations in at least 5% of the cells collected. Chromosomal aberrations were detected in all brush specimens taken from tumors that were DNA aneuploid and showed aneusomy. The presence of these aberrations correlated well with the classification "suspicious for malignancy," which was based on Papanicolaou stained slides of the same specimens. In the control group the percentage of chromosomally aberrant cells did not exceed 2%; in addition, no suspiciously malignant cells were observed in this group. CONCLUSIONS: This study reveals that the FISH technique can be applied diagnostically to brush specimens of HNSCC. The presence of chromosomal aberrations in > 5% of the cells in these specimens can be considered as a marker for malignancy. PMID- 9349520 TI - SNARE proteins--why so many, why so few? AB - Both trafficking and secretion critically depend on accurate and specific membrane recognition and fusion. A key step in these processes is the assembly of a complex consisting of a small number of proteins, i.e., the exocytic core complex. In nerve terminals, this set consists of VAMP and synaptotagmin, which reside at membranes of synaptic vesicles, and syntaxin and SNAP-25 at the plasma membrane. In this survey, different secretory systems that depend on the exocytic core proteins are considered. The possibility that specificity in membrane recognition and fusion is achieved by the numerous variants of proteins of the exocytic core is discussed. Variability of the core complex proteins is determined by the complexity of gene families, isoform-specific localization, and posttranslational modifications. Basic biochemical properties depend on specific isoforms, and the possible protein-protein interactions are determined, in turn, by the compatibility of different isoforms. A correlation between specific variants and distinct biochemical or cellular properties is shown. The outcome of this survey is that heterogeneity in secretion may be dictated by the large number of possible combinations of variants of only a few proteins. PMID- 9349521 TI - Molecular cloning and characterization of a lobster G alphaS protein expressed in neurons of olfactory organ and brain. AB - We have isolated from an American lobster (Homarus americanus) olfactory organ cDNA library a clone, lobG alphaS, with >70% identity to mammalian and arthropod G alphaS sequences. In genomic Southern blots, a fragment of lobG alphaS detected only one band, suggesting the lobsters have a single G alphaS gene. In brain and olfactory organ, lobG alphaS mRNA was expressed predominantly in neurons, including many of the neuronal cell body clusters of the brain. G alphaS protein was also expressed broadly, appearing on western blots as a band of 51.8 kDa in brain, eyestalk, pereiopod, dactyl, tail muscle, olfactory organ, and aesthetasc hairs. These results suggest that lobG alphaS plays a role in a wide variety of signal transduction events. Its presence in the olfactory aesthetasc hairs, which are almost pure preparations of the outer dendrites of the olfactory receptor neurons, and the expression of lobG alphaS mRNA in the olfactory receptor neurons of the olfactory organ indicate that lobG alphaS may mediate olfactory transduction. That virtually all ORNs express lobG alphaS mRNA equally predicts that hyperpolarizing odor responses mediated by cyclic AMP are a property of all lobster olfactory receptor neurons. PMID- 9349523 TI - Cloning and expression of a human serotonin 5-HT4 receptor cDNA. AB - Using a combination of library screening and nested PCR based on a partial human serotonin 5-HT4 receptor sequence, we have cloned the complete coding region for a human 5-HT4 receptor. The sequence shows extensive similarity to the published porcine 5-HT4A and rat 5-HT4L receptor cDNA; however, in comparison with the latter, we find an open reading frame corresponding to only 388 amino acids instead of 406 amino acids. This difference is due to a frame shift caused by an additional cytosine found in the human sequence after position 1,154. Moreover, we also found the same additional cytosine in the rat 5-HT4 sequence. We confirmed the occurrence of the sequence by examining this part of the sequence in genomic DNA of 10 human volunteers and in rat genomic DNA. Based on a part of the genomic 5-HT4 receptor sequence that was identified in the cloning process, there seem to be at least two possible splice sites in the coding region of the gene. The human 5-HT4 receptor, transiently expressed in COS-7 cells, showed radioligand binding properties similar to 5-HT4 receptors in guinea pig striatal tissue. [3H]GR 113808 revealed K(D) values of 0.15 +/- 0.01 nM for the human receptor and 0.3 +/- 0.1 nM in the guinea pig tissue. Binding constants were determined for four investigated 5-HT4 antagonists and three agonists, and appropriate binding inhibition constants were found in each case. Stimulation of transfected COS-7 cells with 5-HT4-specific agonists caused an increase in cyclic AMP levels. PMID- 9349522 TI - Chromosomal localization of the myelin-associated oligodendrocytic basic protein and expression in the genetically linked neurological mouse mutants ducky and tippy. AB - The alternatively spliced cDNAs encoding the myelin-associated/oligodendrocytic basic proteins (MOBPs) have recently been identified in rat. The Mobp gene maps to the distal part of mouse chromosome 9 at a region syntenic with the human chromosome 3p22-p21.3. Two nonallelic mouse mutants, tippy and ducky, with severe neurological phenotypes map to the vicinity of the Mobp locus. We therefore tested whether MOBP malfunction could explain the tippy and ducky defects. In tippy mutant animals, MOBP expression and that of other myelin markers were indistinguishable from wild type. The ultrastructure of tippy myelin was shown to be normal. Ducky animals showed a slight reduction of the brain size, most evident in the spinal cord, but normal progress of myelination. Both MOBP and myelin basic protein expression were lowered only regionally in the CNS, but were mostly normal in the anterior parts of the brain. Ultrastructurally, ducky myelin appeared normal. MOBP transcript sizes and the molecular weights of the encoded proteins were shown to be normal in both mutants. Finally, the nucleotide sequence of the abundant MOBP-81 cDNA was determined and compared with tippy and ducky MOBP-81. Wild-type mouse MOBP-81 protein was 99% identical to the rat homologue, and tippy and ducky MOBP-81 were identical to the wild-type sequence. Our results suggest that alterations in the Mobp gene are not the cause for the severe neurological phenotypes of ducky and tippy mice. PMID- 9349524 TI - Subcellular distribution of glyceraldehyde-3-phosphate dehydrogenase in cerebellar granule cells undergoing cytosine arabinoside-induced apoptosis. AB - We have previously shown that cytosine arabinoside (AraC)-induced apoptosis of cerebellar granule cells (CGCs) results in an increase of a 38-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, identified as glyceraldehyde 3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12). Antisense oligonucleotides to GAPDH mRNA afford acutely plated CGCs significant protection against AraC-induced apoptosis. We used differential centrifugation to examine which subcellular components are affected. Treated and untreated cells were sonicated in 0.32 M sucrose and sequentially centrifuged at 1,000, 20,000, and 200,000 g, to obtain crude nuclear, mitochondrial, microsomal, and cytosolic fractions. Western blotting showed that the levels of GAPDH protein were markedly increased in the 1,000- and 20,000-g pellets. The levels in the cytosolic supernatant were decreased dramatically by AraC in acutely plated CGCs but not in cells 24 h after plating. It is noteworthy that although GAPDH protein in the pellet fractions increased, the dehydrogenase activity of GAPDH decreased. Two other dehydrogenases, lactate dehydrogenase (EC 1.1.1.27) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49), were not similarly affected, suggesting that the effect was GAPDH specific. These observations suggest that GAPDH levels change in specific organelles during apoptosis for reasons that are separate from its function as a glycolytic enzyme. The accumulation of GAPDH protein in specific subcellular loci may play a role in neuronal apoptosis. PMID- 9349525 TI - Molecular characterization and differential gene induction of the neuroendocrine specific genes neurotensin, neurotensin receptor, PC1, PC2, and 7B2 in the human ocular ciliary epithelium. AB - The ocular ciliary epithelium is a bilayer of neuroepithelial cells specialized in the secretion of aqueous humor fluid and the regulation of intraocular pressure. In this study, we report on the expression of the regulatory peptide neurotensin (NT) and a set of differentiated neuroendocrine markers including neurotensin receptors (NTrs), the prohormone convertases furin, PC1, and PC2, and the neuroendocrine polypeptide 7B2 in the ciliary epithelium. Using a human cell line, ODM-2, derived from the nonpigmented ciliary epithelium, we demonstrate that (1) NT expression is highly activated by nerve growth factor, glucocorticoid, and activators of adenylate cyclase; (2) NTr expression is up regulated by selective ligand-activated beta2-adrenergic receptor; and (3) PC1 and PC2 expression are up-regulated via distinct signaling transduction pathways. PC1 gene expression is activated by phorbol ester, and PC2 by the same inducers as those of NT expression. A radioimmunoassay for NT detected an NT-like immunoreactivity in human ciliary epithelium and ODM-2 cell extracts, in aqueous humor, and in conditioned culture medium. The results support the view that the entire ciliary epithelium functions as a neuroendocrine tissue, synthesizing, processing, and releasing NT into the aqueous humor where it may exert important physiological functions through autocrine and/or paracrine mechanisms. PMID- 9349526 TI - Expression of neuronal kinesin heavy chain is developmentally regulated in the central nervous system of the rat. AB - The kinesin family of motor proteins comprises at least two isoforms of conventional kinesin encoded by different genes: ubiquitous kinesin, expressed in all cells and tissues, and neuronal kinesin, expressed exclusively in neuronal cells. In the present study, we have analyzed the expression of the two kinesin isoforms by immunochemistry at different stages of development of the rat CNS. We have found that the level of expression of neuronal kinesin is five to eight times higher in developing than in adult rat brains, whereas that of ubiquitous kinesin is only approximately 2.5 times higher in maturing versus adult brains. Moreover, we have studied the distribution of neuronal kinesin by light microscopic immunocytochemistry in the rat brain at different postnatal ages and have found this protein not only to be more highly expressed in juvenile than in adult rat brains but also to show a different pattern of distribution. In particular, tracts of axonal fibers were clearly stained at early postnatal stages of development but were markedly unlabeled in adult rat brains. Our results indicate that the expression of at least one isoform of conventional neuron-specific kinesin is up-regulated in the developing rat CNS and suggest that this protein might play an important role in microtubule-based transport during the maturation of neuronal cells in vivo. PMID- 9349527 TI - The role of glutathione in dopaminergic neuronal survival. AB - An increased production of reactive oxygen species is thought to be critical to the pathogenesis of Parkinson's disease. At autopsy, patients with either presymptomatic or symptomatic Parkinson's disease have a decreased level of glutathione in the substantia nigra pars compacta. This change represents the earliest index of oxidative stress in Parkinson's disease discovered to this point. This study compares the sensitivity of dopaminergic and nondopaminergic neurons in dissociated mesencephalic cultures to the depletion of glutathione. We have found that dopaminergic neurons are more resistant to the toxicity of glutathione depletion than nondopaminergic neurons. The possibility that dopaminergic neurons have a higher baseline glutathione level than nondopaminergic neurons is suggested by measurements of levels of cellular glutathione in a parallel system of immortalized embryonic dopaminergic and nondopaminergic cell lines. We also examined the role of glutathione in 1-methyl 4-phenylpyridinium toxicity. Decreasing the glutathione level of dopaminergic neurons potentiates their susceptibility to 1-methyl-4-phenylpyridinium toxicity, although 1-methyl-4-phenylpyridinium does not deplete glutathione from primary mesencephalic cultures. Our data suggest that although a decreased glutathione content is not likely to be the sole cause of dopaminergic neuronal loss in Parkinson's disease, decreased glutathione content may act in conjunction with other factors such as 1-methyl-4-phenylpyridinium to cause the selective death of dopaminergic neurons. PMID- 9349528 TI - Neonatal oligodendrocytes contain and secrete neuregulins in vitro. AB - The factors that influence the development of oligodendrocyte (OLG) progenitors into mature OLGs remain elusive. Recent evidence has suggested that neu differentiation factor (NDF), which is a member of the neuregulin family of growth factors, influences the development of glial cells, including Schwann cells, astrocytes, and OLGs. Neurons are postulated to be the source of neuregulins, because neurons closely interact with these glial cells during development. In this report, we have identified the mRNA for both isoform families of NDF in cultured neonatal (immature) OLGs. We have also demonstrated that cultured neonatal OLGs contain and secrete NDF protein. These data raise the possibility that NDF could be used in an autocrine/paracrine loop by neonatal OLGs during development for survival, proliferation, and/or differentiation. PMID- 9349529 TI - Characterization of the palmitoylation domain of SNAP-25. AB - SNAP-25 (synaptosomal associated protein of 25 kDa) is a neural specific protein that has been implicated in the synaptic vesicle docking and fusion process. It is tightly associated with membranes, and it is one of the major palmitoylated proteins found in neurons. The functional role of palmitoylation for SNAP-25 is unclear. In this report, we show that the palmitate of SNAP-25 is rapidly turned over in PC12 cells, with a half-life of approximately 3 h, and the half-life for the protein is 8 h. Mutation of Cys to Ser at positions 85, 88, 90, and 92 reduced the palmitoylation to 9, 21, 42, and 35% of the wild-type protein, respectively. Additional mutations of either Cys(85,88) or Cys(90,92) nearly abolished palmitoylation of the protein. A similar effect on membrane binding for the mutant SNAP-25 was observed, which correlated with the degree of palmitoylation. These results suggest that all four Cys residues are involved in palmitoylation and that membrane association of SNAP-25 may be regulated through dynamic palmitoylation. PMID- 9349530 TI - In vitro induction of apoptosis or differentiation by dopamine in an immortalized olfactory neuronal cell line. AB - A new neuronal cell line was generated by transfection of rat olfactory epithelium with immortalizing recombinant oncogene E1A of adenovirus-2. The resulting 13.S.1.24 line of transformed cells expressed an antigenic phenotype of olfactory neuronal progenitors. Addition of dopamine to 13.S.1.24 cultures induced reduction of cell number within 2 days. Two hallmarks of apoptosis were detected in dopamine-treated cultures: internucleosomal DNA fragmentation and nuclear condensation. Dopamine did not alter the cell proliferation rate, as assessed by [3H]thymidine incorporation. Dopamine also stimulated differentiation of surviving 13.S.1.24 cells into bipolar olfactory marker protein-immunoreactive neurons. Time-dependency assessments over 1 week of treatment indicated that apoptosis and differentiation induced by dopamine were concomitant. Both apoptosis and differentiation triggered by dopamine were dose-dependent, half maximal effects being obtained with approximately 10 microM dopamine. Mediation of both effects by dopaminergic D2 receptors was supported by several observations: active dopamine doses in micromolar ranges, quinpirole agonism and eticlopride antagonism, D2-characteristic rank order of potency among the three agonists tested, and specific binding of a selective D2-like radioligand to 13.S.1.24 cells. The present data altogether indicated that dopamine commits immortalized olfactory neuronal cells in vitro either to apoptosis or to olfactory-like differentiation via D2 dopaminergic receptors. PMID- 9349531 TI - Disrupted [Ca2+]i homeostasis contributes to the toxicity of nitric oxide in cultured hippocampal neurons. AB - Nitric oxide (NO) has been shown to be an important mediator in several forms of neurotoxicity. We previously reported that NO alters intracellular Ca2+ concentration ([Ca2+]i) homeostasis in cultured hippocampal neurons during 20-min exposures. In this study, we examine the relationship between late alterations of [Ca2+]i homeostasis and the delayed toxicity produced by NO. The NO-releasing agent S-nitrosocysteine (SNOC; 300 microM) reduced survival by about one half 1 day after 20-min exposures, as did other NO-releasing agents. SNOC also was found to produce prolonged elevations of [Ca2+]i, persisting at 2 and 6 h. Hemoglobin, a scavenger of NO, blocked both the late [Ca2+]i elevation and the delayed toxicity of SNOC. Removal of extracellular Ca2+ during the 20-min SNOC treatment failed to prevent the late [Ca2+]i elevations and did not prevent the delayed toxicity, but removal of extracellular Ca2+ for the 6 h after exposure as well blocked most of the toxicity. Western blots showed that SNOC exposure resulted in an increased proteolytic breakdown of the structural protein spectrin, generating a fragment with immunoreactivity suggesting activity of the Ca2+-activated protease calpain. The spectrin breakdown and the toxicity of SNOC were inhibited by treatment with calpain antagonists. We conclude that exposures to toxic levels of NO cause prolonged disruption of [Ca2+]i homeostatic mechanisms, and that the resulting persistent [Ca2+]i elevations contribute to the delayed neurotoxicity of NO. PMID- 9349532 TI - Protease-activated receptor-2 (PAR-2) is present in the rat hippocampus and is associated with neurodegeneration. AB - Protease-activated receptor-2 (PAR-2) is a seven-transmembrane G protein-coupled receptor that possesses a structure and activation mechanism similar to those of the thrombin receptor. It is activated by low concentrations of trypsin (300 pM) and a synthetic hexapeptide [sequence of serine, leucine, isoleucine, glycine, arginine, leucine (SLIGRL), the rodent PAR-2 "tethered ligand"] representing the first six amino acids following the putative PAR-2 cleavage site. Previous studies have indicated that alpha-thrombin and SFLLRN (synthetic hexapeptide sequence of serine, phenylalanine, leucine, leucine, arginine, asparagine; the human thrombin receptor "tethered ligand") induce neurite retraction and neurotoxicity. Because of the strong similarities between thrombin receptor and PAR-2, we have proposed that PAR-2 may also participate in neurodegeneration. In the present study, we used reverse transcriptase polymerase chain reaction and immunocytochemistry to provide the first evidence that PAR-2 is present in the rat hippocampus. Moreover, we found SLIGRL to be toxic to hippocampal neurons in a concentration-dependent manner (> or = 100 microM). Calcium signaling studies were performed to aid in determining the mechanism by which PAR-2 activation is neurotoxic. PMID- 9349533 TI - Expression and regulation of types I and II inositol 1,4,5-trisphosphate receptors in rat cerebellar granule cell preparations. AB - Previous studies have shown that as rat cerebellar granule cell cultures differentiate in the presence of 25 mM KCl, they "up-regulate" their ability to form inositol phosphates and release Ca2+ from internal stores in response to the activation of phosphoinositidase C-linked muscarinic and metabotropic receptors. Here we show that they simultaneously up-regulate their ability to respond to inositol 1,4,5-trisphosphate (InsP3) by increasing InsP3 receptor (InsP3R) expression. In contrast, if granule cells are maintained at the more physiological KCl concentration of 5 mM, most cells undergo apoptosis, although a significant number survive. The surviving cells, however, express few InsP3Rs, suggesting that an influx of Ca2+ through voltage-dependent channels is required for InsP3R up-regulation. In addition, we have determined that these cultures express two genetically distinct InsP3R types, but that only one, the type I receptor, is expressed in granule cells. Type II receptors are also present but are found exclusively in astrocytes, which are a minor contaminant of granule cell cultures. This segregation of InsP3R types explains a previous observation, showing that the muscarinic agonist carbachol causes the reduction or "down regulation" of type I but not type II InsP3Rs. PMID- 9349534 TI - Alpha2-macroglobulin complexes with and mediates the endocytosis of beta-amyloid peptide via cell surface low-density lipoprotein receptor-related protein. AB - A primary histopathological feature of Alzheimer's disease is the accumulation of beta-amyloid (A beta) in the brain of afflicted individuals. However, A beta is produced continuously as a soluble protein in healthy individuals where it is detected in serum and CSF, suggesting the existence of cellular clearance mechanisms that normally prevent its accumulation and aggregation. Here, we demonstrate that A beta forms stable complexes with activated alpha2 macroglobulin (alpha2M*), a physiological ligand for the low-density lipoprotein receptor-related protein (LRP) that is abundantly expressed in the CNS. These alpha2M*/125I-A beta complexes are immunoreactive with both anti-A beta and anti alpha2M IgG and are stable under various pH conditions, sodium dodecyl sulfate, reducing agents, and boiling. We demonstrate that alpha2M*/125I-A beta complexes can be degraded by glioblastoma cells and fibroblasts via LRP, because degradation is partially inhibited by receptor-associated protein (RAP), an antagonist of ligand interactions with LRP. In contrast, the degradation of free 125I-A beta is not inhibited by RAP and thus must be mediated via an LRP independent pathway. These results suggest that LRP can function as a clearance receptor for A beta via a physiological ligand. PMID- 9349536 TI - Sensitization of G protein-coupled benzodiazepine receptors in the striatum of 6 hydroxydopamine-lesioned rats. AB - The nonselective benzodiazepine (BZ) agonist diazepam is a potent inhibitor of adenylyl cyclase (AC) activity in the rat striatum. To examine this inhibitory action of diazepam further, its effects were examined in 6-hydroxydopamine lesioned animals, which reportedly exhibit sensitization of the striatal AC pathway. As previously observed, inhibition of AC activity by diazepam was biphasic, with the first phase being receptor-mediated, whereas the second phase involves a direct action on the enzyme itself. In the presence of NaCl (120 mM), a marked sensitization to the receptor-mediated inhibitory effect of diazepam on AC activity was observed in striatal membranes of lesioned animals. EC50 values were 10.4 +/- 1.1 and 4.8 +/- 0.9 nM (p < 0.05) for intact and lesioned striata, respectively. An examination of [3H]diazepam binding revealed a significant increase in the density of binding sites in denervated striata, with no change in affinity. A time-dependent increase in [alpha-32P]GTP labeling of two distinct striatal proteins with apparent molecular masses of 40 and 45 kDa, suggestive of the alpha subunits of Gi and Gs, respectively, was observed. There was a significant increase in basal [alpha-32P]GTP binding to both proteins in lesioned striata. In addition, diazepam stimulated [alpha-32P]GTP binding to the 40-kDa protein, especially in lesioned striata. These data indicate that the sensitization of the receptor-mediated inhibitory effect of diazepam on AC activity in denervated striata may involve up-regulation of BZ receptors as well as enhanced functional coupling of these receptors to inhibitory G proteins. PMID- 9349535 TI - Phorbol ester and calcium regulation of corticotrophin-releasing factor receptor 1 expression in a neuronal cell line. AB - We have previously demonstrated that corticotrophin-releasing factor receptor 1 (CRF-R1) mRNA levels can be down-regulated via activation of the cyclic AMP pathway in CATH.a cells, a neuronal cell line. In this study, we show evidence for down-regulation of CRF-R1 mRNA levels via activation of the protein kinase C (PKC) and calcium second messenger pathways. Incubation of CATH.a cells with phorbol 12-myristate 13-acetate (PMA), an activator of PKC, resulted in a time- and concentration-dependent down-regulation of CRF-R1 mRNA levels. Pretreatment with the inactive phorbol ester 4alpha-phorbol failed to influence significantly CRF-R1 mRNA levels. Incubation with carbachol, a cholinergic agonist known to activate PKC and increase intracellular calcium levels via phosphatidylinositol breakdown, also down-regulated CRF-R1 mRNA levels. Intracellular calcium levels were directly increased using A23187, a calcium ionophore, and thapsigargin, a calcium-ATPase inhibitor. Elevation of intracellular calcium content using either A23187 or thapsigargin significantly down-regulated levels of CRF-R1 mRNA. Furthermore, chelation of calcium with EGTA or blockade of voltage-dependent calcium channels with nifedipine inhibited agonist-mediated down-regulation of CRF-R1 mRNA levels. These results indicate that activation of PKC or calcium signal transduction pathways is sufficient to cause down-regulation of CRF-R1 mRNA levels and that calcium is required for agonist-mediated down-regulation of this receptor. PMID- 9349537 TI - Synaptic vesicle recycling in cultured cerebellar granule cells: role of vesicular acidification and refilling. AB - The role of the transvesicular protonmotive force in synaptic vesicle recycling was investigated in cultured cerebellar granule cells. The vesicular V-ATPase was inhibited by 1 microM bafilomycin A1; as an alternative, the pH component of the gradient was selectively collapsed by equilibration of the cells with 10 mM methylamine and monitored with the fluorescent probe Lysosensor Green. Electrical field-evoked exocytosis of D-[3H]aspartate was inhibited by bafilomycin A1 but not by methylamine, indicating that a transvesicular membrane potential rather than pH gradient is required for transmitter retention within vesicles. In contrast, neither compound affected the field-evoked uptake, recycling, or destaining of the vesicle-specific dye FM2-10; thus, vesicles whose lumens were neutral and/or depleted of transmitter could still recycle in the nerve terminal. No exhaustion of D-[3H]aspartate exocytosis was observed when cells were subjected to six consecutive trains of field stimuli (40 Hz/10 s separated by 10 s). In contrast, the release of preloaded FM2-10 was reduced by approximately 50%, with each stimulus indicating that unlabeled vesicles with accumulated D [3H]aspartate were competing with labeled vesicles for exocytosis. As D [3H]aspartate was accumulated rapidly across the vesicle membrane from the large cytoplasmic pool, the transmitter-loaded but unlabelled vesicles may represent refilled recycling vesicles. FM2-10 destaining and D-[3H]aspartate exocytosis were reduced in parallel at low frequencies, challenging a role for transient vesicle fusion. PMID- 9349538 TI - Decreased zinc affinity of amyotrophic lateral sclerosis-associated superoxide dismutase mutants leads to enhanced catalysis of tyrosine nitration by peroxynitrite. AB - Mutations to Cu/Zn superoxide dismutase (SOD) linked to familial amyotrophic lateral sclerosis (ALS) enhance an unknown toxic reaction that leads to the selective degeneration of motor neurons. However, the question of how >50 different missense mutations produce a common toxic phenotype remains perplexing. We found that the zinc affinity of four ALS-associated SOD mutants was decreased up to 30-fold compared to wild-type SOD but that both mutants and wild-type SOD retained copper with similar affinity. Neurofilament-L (NF-L), one of the most abundant proteins in motor neurons, bound multiple zinc atoms with sufficient affinity to potentially remove zinc from both wild-type and mutant SOD while having a lower affinity for copper. The loss of zinc from wild-type SOD approximately doubled its efficiency for catalyzing peroxynitrite-mediated tyrosine nitration, suggesting that one gained function by SOD in ALS may be an indirect consequence of zinc loss. Nitration of protein-bound tyrosines is a permanent modification that can adversely affect protein function. Thus, the toxicity of ALS-associated SOD mutants may be related to enhanced catalysis of protein nitration subsequent to zinc loss. By acting as a high-capacity zinc sink, NF-L could foster the formation of zinc-deficient SOD within motor neurons. PMID- 9349539 TI - Superoxide dismutase catalyzes nitration of tyrosines by peroxynitrite in the rod and head domains of neurofilament-L. AB - Superoxide dismutase (SOD) catalyzes the nitration of specific tyrosine residues in proteins by peroxynitrite (ONOO-), which may be the damaging gain-of-function resulting from mutations to SOD associated with familial amyotrophic lateral sclerosis (ALS). We found that disassembled neurofilament-L (light subunit) was more susceptible to tyrosine nitration catalyzed by SOD in vitro. Neurofilament-L was selectively nitrated compared with the majority of other proteins present in brain homogenates. Assembled neurofilament-L was more resistant to nitration, suggesting that the susceptible tyrosine residues were protected by intersubunit contacts in assembled neurofilaments. Electrospray mass spectrometry of trypsin digested neurofilament-L showed that tyrosine 17 in the head region and tyrosines 138, 177, and 265 in alpha-helical coil regions of the rod domain of neurofilament-L were particularly susceptible to SOD-catalyzed nitration. Nitrated neurofilament-L inhibited the assembly of unmodified neurofilament subunits, suggesting that the affected tyrosines are located in regions important for intersubunit contacts. Neurofilaments are major structural proteins expressed in motor neurons and known to be important for their survival in vivo. We suggest that SOD-catalyzed nitration of neurofilament-L may have a significant role in the pathogenesis of ALS. PMID- 9349541 TI - Glycosylation of acetylcholinesterase forms in microsomal membranes from normal and dystrophic Lama2dy mouse muscle. AB - The distribution and glycosylation of acetylcholinesterase (AChE) forms in vesicles derived from sarcoplasmic reticulum of normal muscle (NMV) were investigated and compared with those from dystrophic muscle vesicles (DMV). AChE activity was similar in NMV and DMV. Most of the AChE in NMV and half in DMV were released with Triton X-100. Asymmetric (A12) and globular hydrophilic and amphiphilic (G4H, G4A, G2A, and G1A) AChE species occurred in NMV and DMV, the lighter forms being predominant. The percentage of G4H and G4A decreased in DMV. A fraction of the AChE that could not be extracted with detergent was detached with collagenase. Most of the detergent-released A12 AChE from NMV and nearly half in DMV failed to bind to Ricinus communis agglutinin (RCA-I). Conversely, the collagenase-detached isoforms bound to RCA, revealing that asymmetric AChE associated with internal membranes or basal lamina differed in glycosylation. Moreover, nearly half of G4A AChE in DMV and a few in NMV bound to RCA. Most of the RCA-unreactive G4A forms in NMV come from sarcolemma. The results indicate that dystrophy induces minor changes in the distribution and glycosylation of AChE forms in internal membranes of muscle. PMID- 9349540 TI - Dexamethasone prevents hypoxia/ischemia-induced reductions in cerebral glucose utilization and high-energy phosphate metabolites in immature brain. AB - We examined the potential importance of dexamethasone-mediated alterations in energy metabolism in providing protection against hypoxic-ischemic brain damage in immature rats. Seven-day-old rats (n = 165) that had been treated with dexamethasone (0.1 mg/kg, i.p.) or vehicle were assigned to control or hypoxic ischemic groups (unilateral carotid artery occlusion plus 2-3 h of 8% oxygen at normothermia). The systemic availability of alternate fuels such as beta hydroxybutyrate, lactate, pyruvate, and free fatty acids was not altered by dexamethasone treatment, and, except for glucose, brain levels were also unaffected. At the end of hypoxia, levels of cerebral high-energy phosphates (ATP and phosphocreatine) were decreased in vehicle- but relatively preserved in dexamethasone-treated animals. The local cerebral metabolic rate of glucose utilization (lCMRgl) was decreased modestly under control conditions in dexamethasone-treated animals, whereas cerebral energy use measured in a model of decapitation ischemia did not differ significantly between groups. The lCMRgl increased markedly during hypoxia-ischemia (p < 0.05) and remained elevated throughout ischemia in dexamethasone- but not vehicle-treated groups, indicating an enhanced glycolytic flux with dexamethasone treatment. Thus, dexamethasone likely provides protection against hypoxic-ischemic damage in immature rats by preserving cerebral ATP secondary to a maintenance of glycolytic flux. PMID- 9349542 TI - Purification and characterization of naturally soluble neuropathy target esterase from chicken sciatic nerve by HPLC and western blot. AB - Neuropathy target esterase (NTE) activity is defined operatively as the paraoxon resistant mipafox-sensitive phenyl valerate esterase activity. A preparation containing a soluble isoform (S-NTE2) has been obtained from sciatic nerve. It was inhibited by the biotinylated organophosphorous ester S9B [1-(saligenin cyclic phospho)-9-biotinyldiaminononane] in a progressive manner showing a second order rate constant of (3.50 +/- 0.26) x 10(6) M(-1) x min(-1) with an I50 for 30 min of 6.6 +/- 0.4 nM. S-NTE2 was enriched 218-fold by gel filtration followed by strong and weak anion-exchange chromatographies in HPLC. In western blots, this enriched sample showed two bands of endogenous biotinylated polypeptides after treating the blots with streptavidin-alkaline phosphatase complex. When the sample was treated with S9B, another biotinylated band was observed with a molecular mass of approximately 56 kDa, which was not seen when the sample had been pretreated with mipafox before the S9B labeling. It was deduced that this band represents a polypeptide (identified as the S-NTE2 protein) that is bound by both mipafox and S9B and that should be responsible for the progressive S9B inhibition. It is possible that S-NTE2 is the target for attack by compounds that promote delayed neuropathy. PMID- 9349543 TI - Transient global ischemia alters NMDA receptor expression in rat hippocampus: correlation with decreased immunoreactive protein levels of the NR2A/2B subunits, and an altered NMDA receptor functionality. AB - We investigated the gene expression levels, the immunoreactive protein prevalence, and the functional activity of N-methyl-D-aspartate (NMDA) receptor complexes at early times after severe global ischemia challenge in rats. The mRNA expression levels for the NR2A and NR2B subunits of NMDA receptors changed to different degrees within different subregions of the hippocampus after reperfusion with respect to sham-operated control. No significant change in expression was observed in the vulnerable CA1 subfield at or before 6 h after challenge for either receptor subunit, although changes in expression in other hippocampal subfields were observed. At 12 and 24 h after challenge, significant decreases in expression for both subunits were found in the vulnerable CA1 subfield, as well as in other hippocampal regions. At the protein level, a significant decrease in the amount of NR2A/NR2B immunoreactivity in the total hippocampus was observed at both 6 and 24 h after reperfusion compared with sham control. Electrophysiological assessment of single-channel NMDA receptor activity in the CA1 subfield indicates that the main conductance state of NMDA receptor channels is maintained 6 h after challenge, although by 18-24 h after challenge, this main conductance state is rarely observed. The NMDA receptor component of the excitatory postsynaptic field potential was found to be significantly diminished from sham control 24 h after challenge, such that only approximately 10% of the sham response remained, but was not significantly altered from sham control at 6 h after challenge. These results indicate that decreases in the expression levels, the immunoreactive protein prevalence, and that alterations in the functionality of NMDA receptors occur in the hippocampus at early times after severe transient global ischemia. PMID- 9349544 TI - Isoform-specific effects of apolipoproteins E2, E3, and E4 on cerebral capillary sequestration and blood-brain barrier transport of circulating Alzheimer's amyloid beta. AB - Cerebral capillary sequestration and blood-brain barrier (BBB) permeability to apolipoproteins E2 (apoE2), E3 (apoE3), and E4 (apoE4) and to their complexes with sA beta(1-40), a peptide homologous to the major form of soluble Alzheimer's amyloid beta, were studied in perfused guinea pig brain. Cerebrovascular uptake of three apoE isoforms was low, their blood-to-brain transport undetectable, but uptake by the choroid plexus significant. Binding of all three isoforms to sA beta(1-40) in vitro was similar with a K(D) between 11.8 and 12.9 nM. Transport into brain parenchyma and sequestration by BBB and choroid plexus were negligible for sA beta(1-40)-apoE2 and sA beta(1-40)-apoE3, but significant for sA beta(1 40)-apoE4. After 10 min, 85% of sA beta(1-40)-apoE4 taken up at the BBB remained as intact complex, whereas free sA beta(1-40) was 51% degraded. Circulating apoE isoforms have contrasting effects on cerebral capillary uptake of and BBB permeability of sA beta. ApoE2 and apoE3 completely prevent cerebral capillary sequestration and blood-to-brain transport of sA beta(1-40). Conversely, apoE4, by entering brain microvessels and parenchyma as a stable complex with sA beta, reduces peptide degradation and may predispose to cerebrovascular and possibly enhance parenchymal amyloid formation under pathological conditions. PMID- 9349545 TI - Low thiamine diphosphate levels in brains of patients with frontal lobe degeneration of the non-Alzheimer's type. AB - We compared the thiamine and thiamine phosphate contents in the frontal, temporal, parietal, and occipital cortex of six patients with frontal lobe degeneration of the non-Alzheimer's type (FNAD) or frontotemporal dementia with five age-, postmortem delay-, and agonal status-matched control subjects. Our results reveal a 40-50% decrease in thiamine diphosphate (TDP) in the cortex of FNAD patients, whereas thiamine monophosphate was increased 49-119%. TDP synthesizing and hydrolyzing enzymes were unaffected. The activity of citrate synthase, a mitochondrial marker enzyme, was decreased in the frontal cortex of patients with FNAD, but no correlation with TDP content was found. These results suggest that decreased contents of TDP, which is essentially mitochondrial, is a specific feature of FNAD. As TDP is an essential cofactor for oxidative metabolism and neurotransmitter synthesis, and because low thiamine status (compared with other species) is a constant feature in humans, a nearly 50% decrease in cortical TDP content may contribute significantly to the clinical symptoms observed in FNAD. This study also provides a basis for a trial of thiamine, to improve the cognitive status of the patients. PMID- 9349546 TI - Effect of sorbinil and ascorbic acid on myo-inositol transport in cultured rat Schwann cells exposed to elevated extracellular glucose. AB - The effect of long-term (2 weeks) exposure to 0-50 mM glucose and 0-1 mM sorbitol on myo-inositol metabolism was studied in cultured rat Schwann cells. Experiments were carried out to determine the effect of sorbinil and ascorbic acid on myo inositol uptake in rat Schwann cells cultured in the presence of increased extracellular glucose or sorbitol. myo-Inositol uptake and its incorporation into phospholipids decreased significantly when cells were grown in > or = 30 mM glucose for a period of 2 weeks. This inhibitory effect was partly blocked by sorbinil, an aldose reductase inhibitor, in a dose-dependent fashion. Significant prevention was achieved with 0.5 and 1 mM sorbinil. Ascorbic acid also prevented the reduction in myo-inositol uptake due to excess extracellular glucose, at 3 and 30 microM concentrations, but not at 300 microM. Neither sorbinil nor ascorbic acid could prevent the alterations in myo-inositol transport in cells exposed to high sorbitol levels for the same period of time. These data suggest that glucose-induced alteration of myo-inositol transport in Schwann cells is mediated, at least in part, via sorbitol accumulation. This myo-inositol transport impairment is prevented by sorbinil and also by ascorbic acid. Ascorbic acid may hold a fresh promise for the treatment/prevention of diabetic neuropathy/complications, at least as an adjunct therapy along with known aldose reductase inhibitors. PMID- 9349547 TI - Regulation of serotonin release in the frontal cortex and ventral hippocampus of homozygous mice lacking 5-HT1B receptors: in vivo microdialysis studies. AB - To assess the involvement of the serotonin receptor subtype 5-HT1B as terminal autoreceptor regulating 5-HT release in mice, we compared basal values and potassium-evoked changes of extracellular 5-HT levels obtained by in vivo microdialysis in two serotoninergic terminal projection areas of conscious wild type mice with those measured in homozygous mutant mice lacking the gene encoding the 5-HT1B receptor. In the frontal cortex and ventral hippocampus, basal and K+ evoked 5-HT release did not differ between the two strains of mice studied. The infusion via reverse microdialysis of the selective 5-HT1B receptor agonist CP 93,129 (500 nM) decreased significantly K+-evoked 5-HT release in the frontal cortex (by -44%) and ventral hippocampus (by -32%) of wild-type mice but had no effect in mutants. In a similar manner, the mixed 5-HT1B-5-HT1D receptor agonist sumatriptan (800 nM) decreased significantly K+-evoked 5-HT release in the frontal cortex (by -46%) of wild-type mice but had no effect in mutants. These results demonstrated that 5-HT1B knockout mice are not as sensitive to full (CP 93,129) and mixed (sumatriptan) 5-HT1B receptor agonists as are wild-type mice. These data provide in vivo evidence that, in mice, 5-HT1B, but not 5-HT1D, autoreceptors inhibit 5-HT release at nerve terminals located in the frontal cortex and ventral hippocampus. PMID- 9349548 TI - Degradation of tau by lysosomal enzyme cathepsin D: implication for Alzheimer neurofibrillary degeneration. AB - The degradation of different isoforms of human recombinant tau (R-tau; T39, T40, and T44) and fetal tau (F-tau) by cathepsin D (CD) was investigated. Gel electrophoresis and Coomassie Blue staining of different R-tau species digested at pH 3.5 showed very little differences in CD susceptibility. Immunoblotting analyses revealed that amino and carboxy termini of tau were cleaved before other regions. F-tau was most vulnerable to proteolysis at both termini. Digestion of R tau with 0.01 unit of CD/ml at pH 3.5 resulted in cleavage between Phe8-Glu9, Met419-Val420, Thr427-Leu428-Ala429, and Leu436-Ala437 as determined by amino acid sequencing and mass spectroscopy (numbering of amino acids was based on T40). With higher concentrations of CD (1 unit/ml), additional sites of digestion were detected between amino acids 34-161, 200-257, and 267-358. The cleavage sites at amino acids 34-161 and 267-358 were observed at pH 3.5, whereas that at amino acids 200-257 was detected at pH 7.0. Our results suggest that CD cleavage of tau could generate tau fragments with intact microtubule binding domains, which could have a role in the pathogenesis of paired helical filaments (PHFs) in Alzheimer's disease. Such proteolysis might also contribute to the changes of PHF phenotype observed in intracellular and extracellular tangles. PMID- 9349549 TI - In vivo electrochemical measurement of the long-lasting release of dopamine and serotonin induced by intrastriatal kainic acid. AB - Intrastriatal injection of the glutamate agonist kainic acid (KA) in rats has been used to produce an animal model to investigate the mechanism of acetylcholine and GABA cell death associated with Huntington's disease. In the present study, the time course of low (10(-5) M) and high (5 x 10(-3) M) concentrations of KA on striatal dopamine and serotonin release was studied in freely moving rats by using in vivo voltammetry. The response to low concentrations of KA varied between animals, either increasing dopamine release during the injection or increasing dopamine and serotonin after the injection for an extended time, suggesting that 10(-5) KA is near the threshold for KA toxicity in the striatum in rats. High concentrations of KA suppressed dopamine release during injection, with both dopamine and serotonin release increasing and remaining elevated for 1-4 and 7-21 days, respectively. KA-induced changes were inhibited by 6-cyano-7-nitroquinoxaline-2,3-dione and bicuculline increased the release of dopamine but not serotonin. These findings suggest that KA-induced changes in dopamine release resulted from a disinhibition of dopamine neurons due to KA-mediated toxicity of striatal GABA neurons. An alternate possibility is that the change in dopamine and serotonin release may have arisen from a functional modification or degeneration of presynaptic terminals. PMID- 9349550 TI - Amidation of beta-amyloid peptide strongly reduced the amyloidogenic activity without alteration of the neurotoxicity. AB - Beta-amyloid accumulates in cerebral deposits in Alzheimer's disease, so to test the correlation between the neurotoxic and fibrillogenic capacity of beta amyloid, we synthesized a peptide homologous to fragment 25-35 of beta-amyloid (beta25-35) and amidated at the C-terminus (beta25-35-NH2). As the amidation strongly reduced the amyloidogenic capacity of beta25-35, we compared its neurotoxic activity in the amidated (beta25-35-NH2) and nonamidated forms. The viability of primary cultures from fetal rat hippocampus was reduced in a dose related manner (10-100 microM) similarly by beta25-35 and beta25-35-NH2, whereas a scrambled peptide, amidated or nonamidated, did not alter the neuronal viability. The neurotoxic activity of beta25-35-NH2 is mediated by apoptosis as demonstrated by morphological and biochemical investigations. Electron microscopy examination of culture media with beta25-35 or beta25-35-NH2 incubated with neuronal cells for 7 days confirmed the high level of fibrillogenic activity of beta25-35 and the almost total absence of fibrils in the solution with beta25-35 NH2. Furthermore, staining with thioflavine S was used to identify amyloid fibrils, and only the cultures exposed to beta25-35 exhibited intense staining associated with neuronal membranes. These data indicate that the neurotoxic activity of the beta-amyloid fragment is independent of the aggregated state of the peptide. PMID- 9349551 TI - Role of aromatic L-amino acid decarboxylase for dopamine replacement by genetically modified fibroblasts in a rat model of Parkinson's disease. AB - Investigations of gene therapy for Parkinson's disease have focused primarily on strategies that replace tyrosine hydroxylase. In the present study, the role of aromatic L-amino acid decarboxylase in gene therapy with tyrosine hydroxylase was examined by adding the gene for aromatic L-amino acid decarboxylase to our paradigm using primary fibroblasts transduced with both tyrosine hydroxylase and GTP cyclohydrolase I. We compared catecholamine synthesis in vitro in cultures of cells with tyrosine hydroxylase and aromatic L-amino acid decarboxylase together versus cocultures of cells containing these enzymes separately. L-DOPA and dopamine levels were higher in the cocultures that separated the enzymes. To determine the role of aromatic L-amino acid decarboxylase in vivo, cells containing tyrosine hydroxylase and GTP cyclohydrolase I were grafted alone or in combination with cells containing aromatic L-amino acid decarboxylase into the 6 hydroxydopamine-denervated rat striatum. Grafts containing aromatic L-amino acid decarboxylase produced less L-DOPA and dopamine as monitored by microdialysis. These findings indicate that not only is there sufficient aromatic L-amino acid decarboxylase near striatal grafts producing L-DOPA, but also the close proximity of the enzyme to tyrosine hydroxylase is detrimental for optimal dopamine production. This is most likely due to feedback inhibition of tyrosine hydroxylase by dopamine. PMID- 9349552 TI - Evidence of increased oxidative damage in both sporadic and familial amyotrophic lateral sclerosis. AB - Some cases of autosomal dominant familial amyotrophic lateral sclerosis (FALS) are associated with mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1), suggesting that oxidative damage may play a role in ALS pathogenesis. To further investigate the biochemical features of FALS and sporadic ALS (SALS), we examined markers of oxidative damage to protein, lipids, and DNA in motor cortex (Brodmann area 4), parietal cortex (Brodmann area 40), and cerebellum from control subjects, FALS patients with and without known SOD mutations, SALS patients, and disease controls (Pick's disease, progressive supranuclear palsy, diffuse Lewy body disease). Protein carbonyl and nuclear DNA 8-hydroxy-2' deoxyguanosine (OH8dG) levels were increased in SALS motor cortex but not in FALS patients. Malondialdehyde levels showed no significant changes. Immunohistochemical studies showed increased neuronal staining for hemeoxygenase 1, malondialdehyde-modified protein, and OH8dG in both SALS and FALS spinal cord. These studies therefore provide further evidence that oxidative damage may play a role in the pathogenesis of neuronal degeneration in both SALS and FALS. PMID- 9349554 TI - A conformation- and phosphorylation-dependent antibody recognizing the paired helical filaments of Alzheimer's disease. AB - Hyperphosphorylated tau (PHF-tau) is the major constituent of paired helical filaments (PHFs) from Alzheimer's disease (AD) brains. This conclusion has been based largely on the creation and characterization of monoclonal antibodies raised against PHFs, which can be classified in three categories: (a) those recognizing unmodified primary sequences of tau, (b) those recognizing phosphorylation-dependent epitopes on tau, and (c) those recognizing conformation dependent epitopes on tau. Recent studies have suggested that the antibodies recognizing primary sequence and phosphorylation-dependent epitopes on tau are unable to distinguish between normal adult biopsy tau and PHF-tau. We now present evidence for a new fourth class of monoclonal antibodies recognizing conformation dependent phosphoepitopes on tau, typified by TG-3, a monoclonal antibody raised to PHFs from AD brain homogenates. Studies using a series of deletional tau mutants, site-directed tau mutants, and synthetic peptides enable the precise epitope mapping of TG-3. Additional studies demonstrate that TG-3 reacts with neonatal mouse tau and PHF-tau but does not recognize adult mouse tau or tau derived from normal human autopsy or biopsy tissue. Further investigation reveals that TG-3 recognizes a unique conformation of tau found almost exclusively in PHFs from AD brains. PMID- 9349553 TI - Cooperative interception of neuronal apoptosis by BCL-2 and BAG-1 expression: prevention of caspase activation and reduced production of reactive oxygen species. AB - Neuronally differentiated PC12 cells undergo synchronous apoptosis when deprived of nerve growth factor (NGF). Here we show that NGF withdrawal induces actinomycin D- and cycloheximide-sensitive caspase (ICE-like) activity. The peptide inhibitor of caspase activity, N-acetyl-Asp-Glu-Val-Asp-aldehyde, was more potent than acetyl-Tyr-Val-Ala-Asp-chloromethyl ketone in preventing NGF withdrawal-induced apoptosis, suggesting an important role for caspase-3 (CPP32) like proteases. We observed a peak of reactive oxygen species (ROS) 6 h after NGF withdrawal. ROS appear to be required for apoptosis, because cell death is prevented by the free radical spin trap, N-tert-butyl-alpha-phenylnitrone, and the antioxidant, N-acetylcysteine. ROS production was blocked by actinomycin D, cycloheximide, and caspase protease inhibitors, suggesting that ROS generation is downstream of new mRNA and protein synthesis and activation of caspases. Forced expression of either BCL-2 or the BCL-2-binding protein BAG-1 blocked NGF withdrawal-induced apoptosis, activation of caspases, and ROS generation, showing that they function upstream of caspases. Coexpression of BCL-2 and BAG-1 was more protective than expression of either protein alone. PMID- 9349555 TI - Serotonin acts as a radical scavenger and is oxidized to a dimer during the respiratory burst of activated microglia. AB - Serotonin (5-HT) is known to be readily oxidized and to act as a scavenger of reactive oxygen species produced, e.g., in the presence of peroxidase and H2O2 or during the respiratory burst of phagocytes. One major oxidation product formed under these conditions, the 5-HT dimer 5,5'-dihydroxy-4,4'-bitryptamine (DHBT), was suggested to have neurotoxic properties and to contribute to neuronal damage in neurodegenerative disorders. It is shown in the present study that the luminol enhanced chemiluminescence signal measured after stimulation of the respiratory burst activity of cultivated rat microglial cells by the addition of phorbol 12 myristate 13-acetate is suppressed by 5-HT in a dose-dependent manner. During this process, 5-HT is oxidized to DHBT. Neither the intraventricular injection of DHBT nor the addition of DHBT to cultured astrocytes, neurons, or PC-12 cells was found to cause measurable cytotoxic effects. It is concluded that extracellular 5 HT locally released from platelets and 5-HT nerve endings at sites of brain damage or inflammation, through its suppressant effect on the release of reactive oxygen species during the respiratory burst of activated microglia, may contribute to attenuate secondary tissue damage in the CNS. PMID- 9349556 TI - High-density lipoprotein aggregated by oxidation induces degeneration of neuronal cells. AB - We have previously reported that high-density lipoprotein (HDL) exhibits antineuritogenic effects on chicken cerebral cells in culture. In the present study, we show the effects of HDLs, oxidized by UV irradiation or heating, on chicken cerebral neurons in culture. Both treatments produced several physical and chemical changes in the HDLs, i.e., formation of lipid peroxides, enlargement of HDL diameters, an increased exposure of the tryptophan groups of the apolipoprotein A-I to a more hydrophilic environment, formation of bityrosines, and cross-linking of apolipoprotein A-I. When these treatments were performed in the absence of EDTA, most of the modifications described above were more intense and HDLs formed a macroaggregate that displays a rosette-like structure. The aggregated HDLs produced neurodegeneration and death when added to both undifferentiated and differentiated cerebral neurons in culture. This process was accompanied by the disorganization of the cellular microtubular cytoskeleton and hyperphosphorylation of the microtubule-associated protein tau. Native HDL or HDLs treated in the presence of EDTA inhibited the neuritogenesis of undifferentiated neurons but did not show any significant effect on the differentiated neurons in culture. The effects on the cellular cytoskeleton and morphology of aggregated HDLs recall those of the fibrillar beta-amyloid peptide. The present results suggest that aggregated HDLs could participate in neurodegeneration associated with oxidative stress in the CNS. PMID- 9349557 TI - Regulation of spontaneous activity of the delta-opioid receptor: studies of inverse agonism in intact cells. AB - Adenylyl cyclase activity was measured following labelling of the cellular ATP pool with [3H]adenine in intact Rat-1 fibroblasts that had been stably transfected to express the murine delta-opioid receptor (clone D2). Basal [3H]cyclic AMP accumulation was low and was increased substantially by the addition of the diterpene forskolin. The synthetic enkephalin D-Ala2,D-Leu5 enkephalin (DADLE) produced strong inhibition of forskolin-amplified [3H]cyclic AMP production, whereas the delta-opioid ligand ICI174864 augmented forskolin amplified adenylyl cyclase activity. Naloxone was unable to mimic the effects of ICI174864, and coincubation of the cells with these two ligands attenuated the effect of ICI174864. The EC50 (9.4 +/- 0.6 x 10(-8) M) for ICI174864 augmentation of forskolin-stimulated adenylyl cyclase was equal to its estimated Ki. Pertussis toxin pretreatment of clone D2 cells prevented both this effect of ICI174864 and the inhibition produced by DADLE. Use of a Cytosensor microphysiometer demonstrated that treatment of clone D2 cells with DADLE increased and that with ICI174864 decreased the basal rate of cellular proton extrusion. By using these two distinct experimental strategies, ICI174864 was shown to function in a manner anticipated for an inverse agonist, demonstrating that such effects can be observed in intact cells and are not restricted to assays performed on membrane preparations. PMID- 9349558 TI - Identification and characterization of a new serotonergic recognition site with high affinity for 5-carboxamidotryptamine in mammalian brain. AB - We analyzed the existence of an additional serotonin (5-HT) receptor subtype, sensitive to 5-carboxamidotryptamine, in the mammalian brain. Radioligand binding studies with [3H]5-HT were carried out in rat, guinea pig, and human brain membranes, in the presence of unlabeled drugs to mask the binding to all known 5 HT receptors, with the exception of 5-HT1E sites. Under these conditions, unlabeled 5-carboxamidotryptamine still showed a biphasic competition curve with a nanomolar affinity component. Saturation studies with 5 [3H]carboxamidotryptamine were carried out in the presence of (+/-)-8-hydroxy-2 (di-n-propylamino)tetralin, mesulergine, and ergotamine, to mask the binding to all receptors known to be labeled by 5-carboxamidotryptamine. These studies showed the existence in cortex and hippocampus from guinea pig and human brain of a remaining binding site with high affinity (pK(D) = 7.8-8.1) and a unique pharmacological profile. 5-HT and 5-carboxamidotryptamine showed nanomolar affinity, whereas 5-methoxytryptamine recognized this binding site with intermediate affinity. Other drugs exhibited low or very low potency in inhibiting this binding. The addition of 5'-guanylylimidodiphosphate significantly reduced the number of binding sites labeled by 5 [3H]carboxamidotryptamine, in the presence of the masking drugs described above, indicating the interaction with a GTP-binding protein. Preliminary autoradiographic studies in human brain appear to indicate that this 5-HT binding site is present in areas such as the globus pallidus, neocortex, and hippocampus, among others. PMID- 9349559 TI - Glutamate uptake stimulates Na+,K+-ATPase activity in astrocytes via activation of a distinct subunit highly sensitive to ouabain. AB - The excitatory amino acid glutamate was previously shown to stimulate aerobic glycolysis in astrocytes by a mechanism involving its uptake through an Na+ dependent transporter. Evidence had been provided that Na+,K+-ATPase might be involved in this process. We have now measured the activity of Na+,K+-ATPase in cultured astrocytes, using ouabain-sensitive 86Rb uptake as an index. L-Glutamate increases glial Na+,K+-ATPase activity in a concentration-dependent manner with an EC50 = 67 microM. Both L- and D-aspartate, but not D-glutamate, produce a similar response, an observation that is consistent with an uptake-related effect rather than a receptor-mediated one. Under basal conditions, concentration dependent inhibition of Na+,K+-ATPase activity in astrocytes by ouabain indicates the presence of a single catalytic site with a low affinity for ouabain (K0.5 = 113 microM), compatible with the presence of an alpha1 isozyme. On stimulation with glutamate, however, most of the increased activity is inhibited by low concentrations of ouabain (K0.5 = 20 nM), thus revealing a high-affinity site akin to the alpha2 isozyme. These results suggest that astrocytes possess a glutamate-sensitive isoform of Na+,K+-ATPase that can be mobilized in response to increased neuronal activity. PMID- 9349560 TI - Molecular dissection of native mammalian forebrain NMDA receptors containing the NR1 C2 exon: direct demonstration of NMDA receptors comprising NR1, NR2A, and NR2B subunits within the same complex. AB - The subunit compositions of the NR1 C2 exon-containing N-methyl-D-aspartate (NMDA) receptors of adult mammalian forebrain were determined by using a combination of immunoaffinity chromatography and immunoprecipitation studies with NMDA receptor subunit-specific antibodies. NMDA receptors were solubilised by sodium deoxycholate, pH 9, and purified by anti-NR1 C2 antibody affinity chromatography. The purified receptor subpopulation showed immunoreactivity with anti-NR1 C2, anti-NR1 N1, anti-NR1 C2', anti-NR2A, and anti-NR2B NMDA receptor antibodies. The NR1 C2-receptor subpopulation was subjected to immunoprecipitation using anti-NR2B antibodies and the resultant immune pellets analysed by immunoblotting where anti-NR1 C2, anti-NR1 C2', anti-NR2A, and anti NR2B immunoreactivities were all found. Quantification of the immunoblots showed that 46% of the NR1 C2 immunoreactivity was associated with the NR2B subunit. Of this, 87% (i.e., 40% of total) were NR1 C2/NR2B receptors and 13% (6% of total) were NR1 C2/NR2A/NR2B, thus identifying the triple combination as a minor receptor subset. These results demonstrate directly, for the first time, the coexistence of the NR2A and NR2B subunits in native NMDA receptors. They show the coexistence of two splice forms of the NR1 subunit, i.e., NR1 C2 and NR1 C2', in native receptors and, in addition, they imply an NMDA receptor subpopulation containing four types of NMDA receptor subunit, NR1 C2, NR1 C2', NR2A, and NR2B, which, in accord with molecular size determinations, predicts that the NMDA receptor is at least tetrameric. These results are the first quantitative study of NMDA receptor subtypes and demonstrate molecular heterogeneity for both the NR1 and the NR2 subunits in native forebrain NMDA receptors. PMID- 9349561 TI - Modulation of GH4 cell cycle via A1 adenosine receptors. AB - Identification and characterization of A1 adenosine receptors (A1Rs) in a tumor cell line derived from rat pituitary (GH4 cells) was performed by ligand binding and immunocytochemistry. Subsequently, the involvement of A1Rs in the regulation of cell proliferation was studied in these cells. The agonist N6-(R) phenylisopropyladenosine (R-PIA) did not modify the number of cultured cells, but it regulated the kinetics of the cell cycle. By means of experiments of pulse and of pulse and chase with bromodeoxyuridine and further labeling with Hoechst 33258, propidium iodide, and/or fluorescein-conjugated antibodies against bromodeoxyuridine, it was demonstrated that R-PIA, via A1Rs, accelerated progression from G0/G1 to S phase and from S to G2/M phase of the cell cycle, whereas the initiation of a new cycle occurred at the same time in treated and untreated cells. As a consequence, R-PIA did not change the total length of the cycle. This is the first description of cell cycle regulation without modification of cell proliferation. Although pertussis toxin blocked the R-PIA induced inhibition of cyclic AMP production in these cells, it did not affect the R-PIA action on the cell cycle. In contrast, cholera toxin mimicked the action of R-PIA. Thus, it is likely that regulation of the cell cycle via A1Rs is mediated by heterotrimeric G proteins different from those that mediate inhibition of adenylate cyclase. Due to the fact that cells in G0/G1 phase were less susceptible to secretory signals, adenosine, in an autocrine manner and by regulating the cell cycle kinetics, may contribute to the modulation of the secretory capacity of pituitary cells. PMID- 9349562 TI - Transient expression of PG-M/versican, a large chondroitin sulfate proteoglycan in developing chicken retina. AB - We previously showed the expression of PG-M/versican in embryonic chicken retina. In this study, we characterized the alternatively spliced forms of PG-M/versican and their developmental regulation to investigate the implication of PG M/versican in neurite outgrowth from retinal cells during development. On day 5, the immunolocalization of PG-M was first observed at the inner surface of neural retina. On day 7, the pronounced staining was observed in the nerve fiber layer and inner plexiform layer where neural networks of ganglion cells were being formed. As the development proceeded, more intensive staining was observed in these layers. The staining peaked on day 14 and then decreased. Northern analysis and western blotting revealed the presence of a single-sized transcript (13 kb) and the PG-M/versican core protein (550 kDa) on day 14, but the absence of any transcripts or protein bands on day 20, indicating a transient expression of PG M+ (VO), the alternatively spliced form with the most abundant sites for the chondroitin sulfate attachment. Taken together, it is likely that PG-M/versican is involved in neurite outgrowth from ganglion cells during retinal development, and antiadhesion activity of its chondroitin sulfate chains may be important for regulation. PMID- 9349563 TI - Ethanol specifically inhibits NMDA receptors with affinity for ifenprodil in the low micromolar range in cultured cerebellar granule cells. AB - The effect of ethanol on the intracellular Ca2+ concentration response to NMDA in rat cerebellar granule cells grown in low or high KCl concentrations has been studied using image analysis. The cells grown in low KCl displayed high sensitivity for glycine. The subtype-selective antagonist ifenprodil inhibited the response with high (in the low micromolar range) and low (in the high micromolar range) potency. Ethanol affected the high-potency component in these cultures. In cells grown in high KCl the glycine sensitivity was lower, and a low potency for ifenprodil (high micromolar) dominated. These cells were not significantly sensitive to ethanol. The results indicate that the component displaying potency for ifenprodil in the low micromolar range with properties of the NR2B subunit is the target for ethanol action on the NMDA receptor. PMID- 9349564 TI - Development and characterization of antibodies against the N terminus of the human dopamine D4 receptor. AB - The human dopamine D4 receptor (hD4R), which has been implicated in human diseases such as schizophrenia and in a personality trait called "novelty seeking," has not yet been characterized at the protein level. Following epitope scanning of the hD4R, we have produced a highly specific monoclonal antibody named DFR1 raised against an amino-terminal peptide in a predicted extracellular region of the receptor. DFR1 decorated recombinant hD4Rs on the surface of intact Chinese hamster ovary (CHO) cells by flow cytometry and fluorescence microscopy and also recognized recombinant hD4.2, hD4.4, and hD4.7 receptor isoforms by western blot analysis. When expressed stably in CHO cells, all three hD4R isoforms contained N-linked glycosylation and showed apparent molecular masses of 48, 55, and 67 kDa for hD4.2, hD4.4, and hD4.7, respectively. DFR1 immunoreactivity representing hD4R protein or dopamine D4 receptor-like antigens was observed in crude membrane extracts of postmortem human brain tissue by immunoblotting. The DFR1 antibody provides a new immunological tool with the potential to further our understanding of the human dopamine D4 receptor protein. PMID- 9349565 TI - The amyloid precursor protein is not enriched in caveolae-like, detergent insoluble membrane microdomains. AB - The amyloid precursor protein may be processed by several different pathways, one of which produces the amyloid beta-peptide betaA4 present in the amyloid plaques characteristic of Alzheimer's disease. A recent report suggested that axonal amyloid precursor protein is present in a membrane fraction "with caveolae-like properties." In the present study we have isolated detergent-insoluble, caveolae like membranes from both mouse cerebellum and the human neuroblastoma cell line SH-SY5Y. Detergent-insoluble membranes from mouse cerebellum retained nearly all of the glycosylphosphatidylinositol-anchored proteins--alkaline phosphatase, 5' nucleotidase, and the F3 protein--while excluding the majority of the plasmalemmal marker protein alkaline phosphodiesterase I. Although the inositol trisphosphate receptor was highly enriched in this detergent-insoluble fraction, neither amyloid precursor protein nor clathrin immunoreactivity could be detected. Similar results were obtained with SH-SY5Y cells, where 5'-nucleotidase activity was enriched at least 30-fold in the detergent-insoluble membranes, but no amyloid precursor protein or clathrin immunoreactivity could be detected. Caveolin could not be detected in microsomal membranes from either mouse cerebellum or SH-SY5Y cells. These observations suggest that amyloid precursor protein is not normally present in detergent-insoluble, caveolae-like membrane microdomains. PMID- 9349566 TI - Isoform-specific up-regulation by ouabain of Na+,K+-ATPase in cultured rat astrocytes. AB - There are two alpha-subunit isoforms (alpha1 and alpha2) and two beta-subunit isoforms (beta1 and beta2) of Na+,K+-ATPase in astrocytes, but the functional heterodimer composition is not known. Ouabain (0.5-1.0 mM) increased the levels of alpha1 and beta1 mRNAs, whereas it decreased those of alpha2 and beta2 mRNAs in cultured rat astrocytes. The increases in alpha1 and beta1 mRNAs were observed at 6-48 h after addition of the inhibitor. Immunochemical analyses showed that ouabain increased alpha1 and beta1, but not alpha2 and beta2, proteins, and that the isoforms in control and ouabain-treated cultures were of glial origin. Low extracellular K+ and monensin (20 microM) mimicked the effect of ouabain on alpha1 mRNA. The ouabain-induced increase in alpha1 mRNA was blocked by the protein synthesis inhibitor cycloheximide (10 microM), the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetraacetoxymethyl ester (30 microM), and the calcineurin inhibitor FK506 (1 nM). These findings indicate that chronic inhibition of Na+,K+-ATPase up-regulates the alpha1 and beta1, but not alpha2 and beta2, isoforms in astrocytes, suggesting a functional coupling of alpha1beta1 complex. They also suggest that intracellular Na+, Ca2+, and calcineurin may be involved in ouabain-induced up-regulation of the enzyme in astrocytes. PMID- 9349567 TI - Involvement of L-type-like amino acid transporters in S-nitrosocysteine stimulated noradrenaline release in the rat hippocampus. AB - Nitrogen oxides, such as nitric oxide, have been shown to regulate neuronal functions, including neurotransmitter release. We investigated the effect of S nitroso-L-cysteine (SNC) on noradrenaline (NA) release in the rat hippocampus in vivo and in vitro. SNC stimulated [3H]NA release from prelabeled hippocampal slices in a dose-dependent manner. SNC stimulated endogenous NA release within 30 min to almost five times the basal level in vivo (microdialysis in freely moving rats). In a Na+-containing Tyrode's buffer, SNC-stimulated [3H]NA release was inhibited 30% by the coaddition of L-leucine. In the Na+-free, choline-containing buffer, SNC-stimulated [3H]NA release, which was similar to that in the Na+ containing buffer, was inhibited markedly by L-leucine, L-alanine, L-methionine, L-phenylalanine, and L-tyrosine. The effects of the other amino acids examined were smaller or very limited. The effect of L-leucine was stronger than that of D leucine. A specific inhibitor of the L-type amino acid transporter, 2 aminobicyclo[2.2.1]-heptane-2-carboxylate (BCH), inhibited the effects of SNC on [3H]NA release in the Na+-free buffer. Uptake of L-[3H]leucine into the slices in the Na+-free buffer was inhibited by SNC, BCH, and L-phenylalanine, but not by L lysine. The effect of SNC on cyclic GMP accumulation was not inhibited by L leucine, although SNC stimulated cyclic GMP accumulation at concentrations up to 25 microM, much less than the concentration that stimulates NA release. These findings suggest that SNC is incorporated into rat hippocampus via the L-type like amino acid transporter, at least in Na+-free conditions, and that SNC stimulates NA release in vivo and in vitro in a cyclic GMP-independent manner. PMID- 9349568 TI - Phosphorylation of the casein kinase II domain of B-50 (GAP-43) in rat cortical growth cones. AB - Growth-associated phosphoprotein B-50 is a neural protein kinase C (PKC) substrate enriched in nerve growth cones that has been implicated in growth cone plasticity. Here we investigated whether B-50 is a physiological substrate for casein kinase II (CKII) in purified rat cortical growth cone preparations. Using site-specific proteolysis and known modulators of PKC, in combination with immunoprecipitation, mass spectrometry, and phosphoamino acid analysis, we demonstrate that endogenous growth cone B-50 is phosphorylated at multiple sites, on both serine and threonine residues. Consistent with previous reports, stimulation of PKC activity increased the phosphorylation of only those proteolytic fragments containing Ser41. Under basal conditions, however, phosphorylation was predominantly associated with fragments not containing Ser41. Mass spectrometry of tryptic digests of B-50, which had been immunoprecipitated from untreated growth cones, revealed that in situ phosphorylation occurs within peptides B-50(181-198) and B-50(82-98). These peptides contain the major and minor in vitro CKII phosphosites, respectively. In addition, cyanogen bromide digestion of immunoprecipitated chick B-50 generated a 4-kDa C-terminal B-50 phosphopeptide, confirming that phosphorylation of the CKII domain occurs across evolutionary diverse species. We conclude that B-50 in growth cones is not only a substrate for PKC, but also for CKII. PMID- 9349569 TI - Differential regulation of neuronal nicotinic receptor binding sites following chronic nicotine administration. AB - Chronic nicotine administration to rats produces an increase in neuronal nicotinic receptors in the CNS. Moreover, the up-regulated sites labeled by [3H]cytisine in cerebral cortex appear to be composed exclusively of alpha4 and beta2 subunits. It is unknown whether receptor subtypes that do not bind [3H]cytisine with high affinity are also affected. In the present studies, we tested the hypothesis that nicotine treatment differentially alters the density of neuronal nicotinic receptor subtypes in rat nervous tissues. Thus, we compared the binding of [3H]cytisine with that of [3H]epibatidine to nicotinic receptors in brain, spinal cord, and adrenal gland from rats that had been injected twice daily with nicotine or saline vehicle for 10 days. Chronic nicotine treatment led to an increase in nicotinic receptor binding sites in the cerebral cortex and in the dorsal lumbar spinal cord, but not in the thalamus. It is important that virtually all of the observed increases could be accounted for by a selective effect on the fraction of receptors exhibiting high affinity for both [3H]cytisine and [3H]epibatidine. In contrast, no change in [3H]epibatidine binding was seen in the adrenal gland, a tissue that does not exhibit high affinity [3H]cytisine binding. These data indicate that, under the conditions used here, nicotine up-regulates the alpha4beta2 nicotinic receptor subtype, which can be labeled by [3H]cystisine and [3H]epibatidine, but not non alpha4beta2 subtypes, which can be labeled by [3H]epibatidine. PMID- 9349570 TI - Increased hippocampal 5-lipoxygenase mRNA content in melatonin-deficient, pinealectomized rats. AB - Melatonin is neuroprotective because of its antioxidative action, but it also can modify neuronal vulnerability by altering gene expression. 5-Lipoxygenase (5-LO) gene expression is suppressed by the binding of melatonin to its high-affinity nuclear receptors. Recently, we reported that in rats the melatonin deficiency elicited by pinealectomy increases hippocampal susceptibility to excitotoxic injury. Here we have hypothesized that pinealectomy may increase hippocampal vulnerability by eliminating the tonic inhibitory action of melatonin on 5-LO gene expression. Sham-pinealectomized controls and pinealectomized rats were killed 15 days after surgery. Their hippocampi were dissected, and total RNA was extracted and processed for quantitative reverse transcription-polymerase chain reaction assay of 5-LO and cyclophilin mRNAs. Mutated primers were used as internal standards to assay attomole quantities of these two specific mRNAs per microgram of total RNA; the ratio 5-LO/cycophilin was used to compare samples from control and pinealectomized rats. Pinealectomy increased hippocampal 5-LO mRNA content by about threefold. These results support our hypothesis that melatonin deficiency may abate the tonic inhibition of 5-LO mRNA expression and thereby up-regulate 5-LO gene expression, which in turn would increase the brain's synthesis rate of potentially harmful eicosanoids, leukotrienes. PMID- 9349571 TI - Levothyroxine suppressive therapy is partially effective in treating patients with benign, solid thyroid nodules and multinodular goiters. AB - We prospectively evaluated the effect of thyrotropin (TSH)-suppressive therapy with levothyroxine (LT4) on the size of a benign, solitary, solid nodule and multinodular goiter in a relatively low iodine intake area. In this study, 101 euthyroid subjects with a benign, solitary, predominantly solid nodule (n = 54) confirmed by biopsy or multinodular goiter (n = 47) received 200 microg of levothyroxine daily as a single morning tablet for 12 months. Thirty-five receiving no therapy were considered as controls (solitary nodules, n = 20, multinodular, n = 15). Patients were admitted to the study after evaluation of thyroid biochemical parameters (thyroxine [T4], free thyroxine [FT4], triiodothyronine [T3], thyrotropin [TSH], and thyroglobulin [Tg]), thyroid scanning, ultrasound examination, and fine-needle aspiration biopsy. Every 3 months, thyroid function tests and every 6 months ultrasound examinations were repeated. Twelve months later 20 of 54 (37.1%) patients with single, solid nodules had 50% or more regression of the nodular volume (responders). Eleven of 54 (20.3%) patients had more than 20%, but less than 49.9% reduction of nodular volume (partial responders). Nonresponders were 23 of 54 (42.5%). One-third of subjects with multinodular goiter had 50% or more regression of the glandular volume, whereas 46.8% were considered as nonresponsive. The mean serum Tg levels decreased significantly only in responders with solitary nodular disease or multinodular goiter. In the control group only 1 patient (5% of total) with a solitary nodule had a 50% reduction in the nodular volume. Five others had a partial response (<49%, >20% reduction). None of the patients with multinodular goiter had a significant reduction (>50%) of the combined nodular volumes. We concluded that LT4 may be effective, among other factors, in arresting the growth or in reducing the volume of relatively small, benign, solitary, solid thyroid nodules or the combined nodular volume of multinodular goiter. PMID- 9349572 TI - Is percutaneous ethanol injection a useful alternative for the treatment of the cold benign thyroid nodule? Five years' experience. AB - We describe our 5-year experience with percutaneous ethanol injection (PEI) for the treatment of cold benign thyroid nodules and report its efficacy and side effects. Fifty-four euthyroid outpatients (aged 44.8+/-12.7 years, mean+/-SD) were divided into two groups matched for sex, age, and nodule volume: 27 patients treated only by PEI and 27 patients treated additionally with levothyroxine suppressive therapy (median follow-up: 24 months, range 6-48). Mean pretreatment nodule volume was 21.0 mL (range 5.4-54.6). Ethanol (1.3+/-0.6 mL/mL nodule volume) was injected under sonographic control in 4 to 13 weekly sessions (mean 7.4). PEI therapy was well tolerated by all patients. At the end of treatment, nodule volume was 7.7+/-5.7 mL (p = .0001). A further significant shrinkage was obtained at 1-year follow-up (4.4+/-3.8 mL; p < .05). No significant differences in nodule reduction were observed between the levothyroxine treated or untreated group and between patients with pretreatment nodule volume smaller or larger than 15 mL. Our study confirms the efficacy and safety of PEI in inducing volume shrinkage of cold benign thyroid nodules. Overall our data suggest that PEI may become an interesting alternative for patients with surgical indications, if they refuse surgery or are poor surgical risks, or eventually demand treatment for aesthetic purposes. It may also be considered when levothyroxine therapy is contraindicated or ineffective. PMID- 9349573 TI - Value of combined technetium-99m hydroxy methylene diphosphonate and thallium-201 imaging in detecting bone metastases from thyroid carcinoma. AB - Detectability of bone metastases from differentiated thyroid carcinoma by technetium-99m hydroxymethylene diphosphonate ([99m]Tc-HMDP) bone scan is considered to be poor. Thallium-201 (201Tl) is also widely used for detecting metastatic lesions. Our present study was aimed at the evaluation of the combined use of (99m)Tc-HMDP and 201Tl imaging in successful detection of bone metastases from differentiated thyroid carcinoma. Twenty-seven thyroidectomized thyroid cancer patients (19 females, 8 males; 12 papillary type, 15 follicular type) with 77 bone lesions were included in this retrospective study. All of these patients received ablative doses of radioiodine. Thyroidal origin of the lesions was proved by positive iodine-131 (131I) uptake. In 131I-negative lesions, histological proof or absence of tumor markers other than thyroglobulin was considered when computed tomography (CT) and/or magnetic resonance imaging (MRI) suggested metastatic nature of the lesions. Of the 77 lesions, 58 (75.3%) were positive and 19 were negative in the (99m)Tc-HMDP bone scintigraphy, whereas 53 lesions (68.9%) could be detected by 201Tl scintigraphy. However, within the 19 (99m)Tc-HMDP-negative lesions, 14 showed abnormal accumulation of 201Tl, and within the 24 201Tl negative lesions, 19 were positive in (99m)Tc-HMDP scan. This resulted in a combined sensitivity of 93.5%. Our present study concludes that combined (99m)Tc-HMDP and 201Tl imaging is a sensitive and effective method for detecting bone metastases from thyroid carcinoma. PMID- 9349574 TI - Human thyroid carcinoma cell lines and normal thyrocytes: expression and regulation of matrix metalloproteinase-1 and tissue matrix metalloproteinase inhibitor-1 messenger-RNA and protein. AB - Matrix metalloproteinase-1 (MMP-1) and tissue matrix metalloproteinase inhibitor 1 (TIMP-1) play an important role in remodeling the extracellular matrix in normal and pathological processes. The effect of phorbol-myristate acetate (PMA), interleukin-1 (IL-1), and tumor necrosis factor-alpha (TNF-alpha) on MMP-1 and TIMP-1 expression was studied on highly purified thyrocytes and undifferentiated 8505 C, C 643, HTh 74, SW 1736 thyroid carcinoma cells compared with thyroid derived fibroblasts. Messenger RNA (mRNA) levels were monitored by competitive semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) after 24 hours. Culture supernatants were assayed for free and/or complexed MMP-1 and TIMP-1 after 48 hours using enzyme-linked immunosorbent assay (ELISA) systems (detection limit: <2 ng/mL). MMP-1 and TIMP-1 mRNA were present in all cell types, although thyrocytes showed MMP-1 mRNA levels near the detection limit. 8505 C expressed MMP-1 mRNA levels of up to 10(6) times those of the other cells analyzed. PMA and IL-1 increased MMP-1 mRNA in most cell types. TIMP-1 mRNA increased after treatment with PMA in all cells except 8505 C, whereas only slight effects were shown after IL-1 stimulation. MMP-1 protein was undetectable in normal thyrocyte cultures, but was secreted spontaneously by all cell lines ([ng/mL]; C 643: 15+/-7; HTh 74: 81+/-1; SW 1736: 13+/-2; 8505 C: 2097+/-320). There was a strong correlation between levels of MMP-1 mRNA and protein (r = 0.99, p < .0001). PMA and IL-1 increased MMP-1 secretion in all cell types after 48 hours. Fibroblasts ([ng/mL] 517+/-55) and the cell lines (C 643: 142+/-48; HTh 74: 115+/-13; SW 1736: 202+/-14; 8505C: 120+/-19) secreted TIMP-1 in unstimulated cultures, whereas only a trace amount was detected in thyrocyte cultures, even after PMA treatment. IL-1 upregulated TIMP-1 secretion after 48 hours in SW 1736, HTh 74, and C 643 cells. Our data suggest that in contrast to normal thyrocytes, dedifferentiated thyroid carcinoma cell lines are potential producers of MMP-1 as well as TIMP-1. High MMP-1 or MMP-1/TIMP-1 expression may play a role in tissue invasion of undifferentiated thyroid cancer cells. PMID- 9349575 TI - MUC1 mucin gene, transcripts, and protein in adenomas and papillary carcinomas of the thyroid. AB - MUC1 mucin is found in a variety of epithelial tissues and is overexpressed in several epithelial cancers. This molecule could modulate cellular adhesion and thereby influence tumor invasion and metastasis. Little is known of MUC1 gene expression in thyroid tissues. We investigated whether MUC1 mucin gene alteration and/or expression correlated with thyroid tumor progression by studying 21 fresh thyroid tissue specimens comprising 10 macrofollicular adenomas and 11 papillary carcinomas. Normal adjacent tissue from the same patients was also studied. To determine the integrity and expression of the MUC1 mucin gene, a complementary DNA (cDNA) probe was used for Southern blot analysis of DNA and Northern blot analysis of RNA. A detailed immunohistochemical analysis of MUC1 protein expression was performed with DF3 monoclonal antibody, and was compared with other tumor characteristics and clinical manifestations at diagnosis. Of the 14 tumors informative (heterozygous) with the pMUC10 polymorphic probe, 2 (14%) showed loss of heterozygosity (1 adenoma and 1 carcinoma). Overexpression of MUC1 RNA, compared with normal thyroid tissue, was observed in 6 of the 11 papillary carcinomas and in none of the 10 adenomas. Immunostaining of the corresponding formalin-fixed paraffin-embedded tissue sections detected MUC1 mucin protein at the apical domain of follicular cells. Most of the lining was thin in normal tissue and follicular adenomas, but the protein was more irregularly and less strongly expressed in adenomas. In carcinomas the epithelial mucin produced by the MUC1 gene was present irregularly as a thin and/or thick lining at the apical domain of tumor cells. In addition, 5 of the 6 samples with MUC1 mRNA overexpression showed intracytoplasmic staining. Moreover, intracytoplasmic MUC1 mucin staining was found in 75% of "high-risk" papillary thyroid carcinoma (PTC) (PTC with extrathyroid extension at initial diagnosis and/or lymph node involvement), and in only 28.5% of "low-risk" PTC (purely intrathyroidal carcinomas). PMID- 9349576 TI - Incidence and clinical characteristics of thyroid carcinoma after iodine prophylaxis in an endemic goiter country. AB - Iodized salt prophylaxis has been performed in Austria since 1963. Through this approach, mean urinary iodine excretion has been normalized to 144+/-23.5 microg/g creatinine per day. Thus Tyrol is no longer an endemic goiter area. We have analyzed the impact of iodized salt prophylaxis on thyroid cancer (TC) comparing data from the early 1960s with those corresponding to the period 1986 to 1995, when iodine supply was normalized. The study included 439 patients from Tyrol and Southern Tyrol. The incidence of TC in Tyrol has risen during the past decades from 3.07 between in 1957 and 1970 to 7.8 between 1990 and 1994 (CR/100000/year). We observed a rise in the percentage of differentiated adenocarcinomas (56% to 91.5%) with a predominance of papillary TC (54.4%) along with a decrease of anaplastic TC. In addition to these histological features, a shift to less advanced TNM stages, eg, T1-3, N0-1a, M0, was obvious, increasing from 29% to 72.2%, whereas advanced tumors, ie, T4 or N1b or M1, decreased from 71% to 28%. These changes have significantly improved prognosis. The current 5 year survival rate is 90.7% as compared with a rate of 73% in the 1960s; the values for 7-year survival are 89% and 48%, respectively. The marked effects of age, tumor stages, and histology on prognosis were confirmed with the Kaplan Meier method. We conclude that together with normalization of iodine supply in an endemic goiter region the epidemiological profile of TC has changed. Even though the incidence of TC has risen, prognosis has significantly improved due to a shift towards differentiated forms of TC that are diagnosed at earlier stages. PMID- 9349577 TI - Opposite changes in serum soluble CD8 in patients at the active stages of Graves' and Hashimoto's diseases. AB - Serum concentrations of soluble CD8 (sCD8) were examined by enzyme immunoassay in 154 patients with autoimmune thyroid diseases and 46 healthy controls. The numbers of peripheral CD8+ cells were also examined in the same subjects by flow cytometry. The serum concentrations of sCD8 were increased in patients with stimulative thyrotoxicosis caused by active Graves' disease, and decreased in patients with transient destructive thyrotoxicosis caused by the aggravation of Hashimoto's disease, and normal in euthyroid and hypothyroid patients with Graves' or Hashimoto's disease. The ratios of serum sCD8 levels to the numbers of CD8+ cells were increased in thyrotoxic patients with active Graves' disease, but not with active Hashimoto's disease, suggesting an increase in sCD8 production by CD8+ cells in active Graves' disease. The serum concentrations of sCD8 were correlated with the serum levels of thyrotropin receptor antibody (TRAb) and thyroid hormones in Graves' disease. These data indicate that serum sCD8 proteins change in opposite directions in the active stages of Graves' and Hashimoto's diseases, and may represent the disease activities. PMID- 9349578 TI - Urinary iodine excretion during normal pregnancy in healthy women living in the southwest of France: correlation with maternal thyroid parameters. AB - A prospective study was undertaken to evaluate urinary iodine excretion and changes of maternal thyroid function during pregnancy in healthy women living in the southwest of France. The cohort included a total of 347 pregnant women (mean age 28.0+/-0.5 years). Iodine concentration in a random urine sample and thyroid tests (free thyroxine [FT4], free triiodothyronine [FT3], thyrotropin (TSH), thyroxine-binding globulin [TBG], and thyroglobulin [Tg]) were measured at initial presentation (before 12 weeks of gestation), and during the ninth month of pregnancy. A thyroid ultrasound was performed 1 to 5 days after delivery in 246 mothers. Mean urinary iodine levels were low during the first trimester (6.9+/-0.4 microg/dL), as well as during the ninth month of pregnancy (8.6+/-0.6 microg/dL). During pregnancy, FT4 and T3 concentrations decreased (p < .001), and TSH and Tg concentrations increased (p < .001). Thyroid hypertrophy (thyroid volume greater than 18 mL) was present in 15.4% of women whose first trimester urinary iodine concentration was less than 5 microg/dL, but was present in only 3.5% of women whose urinary iodine concentration was greater than 10 microg/dL. A goiter (thyroid volume greater than 22 mL) was present in 11% of the mothers. In conclusion, this prospective study shows that urinary iodine excretion is low in pregnant women living in the southwest of France. This low iodine intake is associated with reduced circulating thyroid hormone levels and growth of the thyroid gland. These data point to the need of an increased iodine supply in these pregnant women to reduce the potential consequences of low iodine intake on maternal thyroid economy. PMID- 9349579 TI - Autoimmune hyperthyroidism in prepubertal children and adolescents: comparison of clinical and biochemical features at diagnosis and responses to medical therapy. AB - We explored our clinical impression that young children with autoimmune hyperthyroidism are more thyrotoxic at presentation and require a longer course of medical therapy than do adolescents to achieve remission. A retrospective chart review of clinical and biochemical data at presentation and response to therapy in 32 prepubertal (PREPUB) and 68 pubertal (PUB) children and adolescents with autoimmune hyperthyroidism was undertaken. Initial therapy included prophylthiouracil or methimazole in all but 11 patients who chose radioactive iodine (131I); 30 additional patients ultimately chose 131I or surgery after an initial period of medical therapy. In PREPUB children there were significantly longer duration of symptoms (7.8+/-7.7 months) and higher serum concentrations of triiodothyronine (T3) 708+/-330 ng/dL) at presentation than in the PUB group (4.7+/-3.4 months; p < .05) (537+/-197 ng/dL; p < .01). Duration of symptoms correlated negatively with chronologic age (r = -0.24; p < .02) but not with T3 or thyroxine (T4) levels (p = .1). PUB children had significantly higher titers of thyroid microsomal antibodies (positive dilution factor 1:6022+/-14572) than did PREPUB children (1:592+/-1226; p < .05). There was a higher familial incidence of thyroid disease in boys (80%) than in girls (64%) (p < .02). The duration of medical therapy was significantly longer (3.5+/-2.9 years) in PREPUB children compared to the PUB group (2.2+/-1.8 years) (p < .05). Only 17% of PREPUB treated 5.9+/-2.8 years compared with 30% of PUB treated 2.8+/-1.1 years achieved a 1-year remission after stopping antithyroid medication (percentage between groups, p < .01; years of treatment, p < .05). The median time to remission after medical therapy was 8 years in PREPUB and 4 years in PUB (p < .02). PREPUB children continued to remit after prolonged medical therapy (>6 years) whereas PUB patients did not. Total treatment length correlated negatively with chronological age (r = -0.26; p < .05) and positively with T4 and T3 concentrations at diagnosis (r = 0.31; p < .01). The diagnosis of hyperthyroidism is delayed in prepubertal children compared to adolescents. This delay may contribute to the higher T3 levels observed in this group at presentation. Prepubertal children also appear to require longer medical therapy to achieve a lower rate of remission, but do continue to remit after prolonged treatment. These differences in response to therapy should be considered when discussing therapeutic options with the family. PMID- 9349580 TI - Brain magnetic resonance imaging in congenital hypothyroid infants at diagnosis. AB - The aim of our study was to investigate the central nervous system (CNS) morphology and myelination with magnetic resonance imaging (MRI) in congenital hypothyroid (CH) infants detected by neonatal screening before replacement therapy. We studied 11 CH infants, 9 girls and 2 boys, mean age 22 days, 3 with aplasia, 5 with ectopia, 2 with hypoplasia of the thyroid gland, 1 with unknown diagnosis. As normal controls 22 term newborns (38 to 41 weeks of gestational age) were studied. MRI studies were performed with a 1.5-T magnet, extremity coil, T1-weighted and heavily T2-sequences axial sections were obtained. No sedation was needed for the MRI studies. MRI brain examination was normal in all patients compared with controls. In particular, no differences in the myelination patterns of the brain were observed between normal subjects and patients with hypothyroidism. Our study shows no morphological brain abnormalities in CH infants detected by neonatal screening before replacement therapy. Perinatal hypothyroidism seems to have no effect on CNS structures. PMID- 9349581 TI - Congenital nonautoimmune hyperthyroidism in a nonidentical twin caused by a sporadic germline mutation in the thyrotropin receptor gene. AB - Congenital hyperthyroidism is usually caused by maternal-to-fetal transfer of thyroid-stimulating antibodies from a mother with autoimmune thyroid disease. Very recently, activating thyrotropin (TSH) receptor germline mutations were detected in a few patients with sporadic nonautoimmune congenital hyperthyroidism, as well as in familial forms of nonautoimmune hyperthyroidism defining a new pathophysiological entity of hyperthyroidism. In this report, we describe a nonidentical twin girl with severe congenital hyperthyroidism. The twin brother and the mother were euthyroid. Skull radiographs revealed premature synostosis of the sagittal sutures. Hyperthyroidism was inadequately controlled with antithyroid drugs and radioiodine therapy. After a near-total thyroidectomy performed at age 3, the patient became hypothyroid and required thyroid hormone replacement. At age 14, hyperthyroidism recurred. A hyperplastic remnant of the right upper lobe was removed surgically, resulting in euthyroidism. Over the following years, thyroid hormone levels increased gradually and at age 19 she was again hyperthyroid. There was no clinical or biochemical evidence of an autoimmune process. The patient's neurologic development was impaired and her intelligence is subnormal. Direct sequencing of the TSH receptor gene revealed a heterozygous mutation resulting in a substitution of threonine632 by isoleucine in the sixth transmembrane segment, an amino acid change known to result in constitutive activation of the cyclic adenosine monophosphate (cAMP) pathway. The mutation was absent in the parents and the twin brother, indicating a de novo germline mutation. Early recognition of this disorder is important because of the resistance to standard treatment, special therapeutic implications, and the possibility of familial transmission. PMID- 9349582 TI - A novel point mutation of thyroid hormone receptor beta gene in a family with resistance to thyroid hormone. AB - Resistance to thyroid hormone (RTH) is characterized by variable tissue hyporesponsiveness to thyroid hormone caused by mutations of thyroid hormone receptor beta (TRbeta) gene. We found a novel point mutation of the TRbeta gene in a family (F123) with RTH, a transition of a guanine to adenine at nucleotide 1215, which replaced the normal Met-310 with Ile. This substitution was found in only one allele of affected family members. In vitro transcription and translation of this mutant TRbeta demonstrated a 12-fold reduction of the affinity for triiodothyronine (T3) compared with the wild type TRbeta. Thyroid function tests were similar to a previously reported RTH family (F99) who had a different mutation in the same codon (Thr 310). PMID- 9349583 TI - Clinical and hormonal outcome after two years of triiodothyroacetic acid treatment in a child with thyroid hormone resistance. AB - We report here a new family with thyroid hormone resistance (RTH), with phenotypic variability among subjects. Particular emphasis is given to the clinical and hormonal outcome after 2 years of triiodothyroacetic acid (TRIAC) treatment in an affected child with peripheral thyrotoxic features (pituitary RTH [PRTH]). The genetic defect was a substitution in position 1642 (C to A) within the exon 10 of thyroid hormone receptor beta1 (TRbeta1) gene, resulting in the codon change P453T. The mutant receptor had a significantly reduced triiodothyronine (T3) binding affinity. Within this family, the child and the mother suffered from hyperthyroidism and were clinically classified as PRTH, while the maternal grandmother was clinically euthyroid, indicating a generalized form of the disease (GRTH). Rapid normalization of heart rate was initially obtained by the association of the cardioselective beta-blocker atenolol with TRIAC. Nevertheless, long-term TRIAC therapy, through its lowering action of serum thyrotropin (TSH) and thyroid hormone levels, maintained a normal heart rate after atenolol discontinuation and normalized the neurological disturbances and the clinical signs in the child, without any apparent side effect. In fact, growth velocity remained unchanged and no alteration of several parameters of thyroid hormone action at the tissue level was observed, whereas soluble interleukin-2 receptor levels improved significantly, confirming the safety and efficacy of long-term TRIAC therapy for PRTH also during childhood. We thus recommend testing the efficacy of TRIAC therapy in all RTH patients presenting with clinical features of hyperthyroidism. PMID- 9349584 TI - Paracrine effect of human chorionic gonadotropin ectopically produced from papillary thyroid cancer cells on growth and function of FRTL-5 rat thyroid cells. AB - It is well known that human chorionic gonadotropin (hCG) is sometimes secreted from nontrophoblastic neoplasms. To elucidate the role of ectopic hCG, we investigated the effect of hCG produced from a papillary thyroid cancer cell line (B-CPAP cells) on stimulation and growth promotion of FRTL-5 rat thyroid cells. Ectopic hCG contained in the culture medium of B-CPAP cells was purified using gel filtration and bioassayed for thyrotropic activity in FRTL-5 cells. Addition of ectopic hCG (up to 5.2 x 10(4) IU/L) increased cyclic adenosine monophosphate (cAMP) accumulation and 3H-thymidine incorporation in FRTL-5 cells dose dependently. These effects were almost as potent as the stimulation induced by standard hCG CR-127. After the absorption of the ectopic hCG by anti-hCG-beta monoclonal antibody, the cAMP accumulation was significantly decreased. Analysis of ectopic hCG isoforms with different isoelectric points indicated the predominance of the acidic hCG isoform with isoelectric point (pI) 3.8-3.2 that is the major isoform of standard hCG. Basic isoforms (pI 5.7-5.3) with higher thyrotropic potency were also detected. These results indicate that the ectopic hCG secreted from papillary thyroid cancer cells possess intrinsic thyroid stimulating and growth-promoting activity. The ectopic hCG may act as an autocrine-paracrine factor in nontrophoblastic neoplasms. PMID- 9349585 TI - Desialylated and deglycosylated human chorionic gonadotropin are superagonists of native human chorionic gonadotropin in human thyroid follicles. AB - Highly purified human chorionic gonadotropin (hCG) interacts with the thyrotropin (TSH) receptor and stimulates triiodothyronine (T3) secretion, iodide uptake and organification, and cyclic adenosine monophosphate (cAMP) formation in human thyroid follicles. Because of interest in the role of the carbohydrate component in the structure-function relationships of hCG we undertook to deplete hCG of its sialic acid or carbohydrate residues and assess the thyrotropic activity of the carbohydrate-modified forms. For this purpose, we used our assay system consisting of human thyroid follicles cultured and suspended in collagen gel in serum-free medium. Under these conditions, the cells are organized as follicular three-dimensional structures with normal polarity, enabling enhanced responsiveness to hormonal stimulation, and T3 secretion can be measured as a response parameter. Desialylated (ds)-hCG and deglycosylated (dg)-hCG dose dependently stimulated T3 secretion, iodide uptake and organification, and in each case did so with about twice the intrinsic activity of native hCG. Indeed, removal of the sialic acid or carbohydrate residues from native hCG transformed it into a thyroid stimulator that elicited a maximal response in terms of iodide uptake, organification and T3 secretion by human thyroid follicles as high as TSH and almost twice as high as native hCG. Not only were ds-hCG and dg-hCG more intrinsically active than hCG, they were more than five times as potent. As with hCG, both ds-hCG and dg-hCG managed to elicit such responses in human thyrocytes while evoking minimal amounts of cAMP, illustrating the concept of cAMP superfluity and highlighting the potential pitfalls of using cAMP as a measure of hormonal bioactivity. hCG, and to a greater extent ds-hCG and dg-hCG, inhibited, as did TSH, gamma-interferon-induced human leukocyte antigen-DR (HLA-DR) expression in human thyrocytes, again reflecting the intrinsic thyrotropic activity of native hCG and its variants depleted of sialic acid or carbohydrate residues. In conclusion, this is the first report on the thyrotropic activity of ds-hCG and dg-hCG using the physiologically relevant hormonal end-point response, thyroid hormone secretion. The study was conducted in a serum-free culture system of human thyroid follicles and shows that removal of the sialic acid or carbohydrate residues from native hCG transform hCG variants into thyroid stimulating superagonists. The hCG variants inhibited, as did TSH, gamma interferon-induced HLA-DR expression. PMID- 9349586 TI - Transforming growth factor-beta1 suppresses thyrotropin-induced Na+/I- symporter messenger RNA and protein levels in FRTL-5 rat thyroid cells. AB - Iodide transport into the thyroid catalyzed by the Na+/I- symporter (NIS), is the first and main rate-limiting step in thyroid hormone synthesis. Recently, we have demonstrated that thyrotropin (TSH) increases NIS messenger RNA (mRNA) and protein levels, as well as iodide uptake activity. Although transforming growth factor-beta1 (TGFbeta1) is known to affect thyroid cell function, it is still unclear how TGFbeta1 regulates TSH-stimulated iodide accumulation. Therefore, the effects of TGFbeta1 on TSH-stimulated NIS mRNA and protein levels were examined in FRTL-5 rat thyroid cells by Northern and Western blot analyses, and iodide uptake was assessed. Northern blot analysis revealed that TGFbeta1 suppressed TSH stimulated NIS mRNA levels in a dose- and time-dependent manner. Western blot analysis demonstrated that TGFbeta1 suppressed TSH-stimulated NIS protein levels. TGFbeta1 also suppressed (Bu)2 cyclic adenosine monophosphate (cAMP)- and forskolin-stimulated NIS mRNA and protein levels, indicating a role for TGFbeta1 downstream of cAMP production. As predicted, TGFbeta1 inhibited TSH-stimulated iodide uptake activity. These results suggest that the inhibitory effect of TGFbeta1 on TSH-stimulated iodide uptake is at least in part due to a suppression of NIS specific transcription. Therefore, TGFbeta1 may act as an autocrine or paracrine local modulator of thyroid hormone synthesis by influencing NIS mRNA levels in the thyroid. PMID- 9349587 TI - Long-term effect of norepinephrine on iodide uptake in FRTL-5 cells. AB - The sympathetic nervous system plays a role in the regulation of thyroid function. In FRTL-5 rat thyroid cells, norepinephrine (NE) acutely depresses intracellular I- by increasing I- efflux. The present study was undertaken to determine the effect of NE on iodide transport after a longer time period. NE inhibited the ability of thyrotropin (TSH) to induce iodide uptake by FRTL-5 cells after 48 or 72 hours, but not after 24 hours. The effect of NE was more evident with increasing concentrations of TSH. NE did not modify the rate of I- efflux. Inhibition was associated with a decrease in the Vmax and no change in the Km for iodide influx. To determine if this was a generalized effect of NE on thyroid cell membrane, the uptake of alpha-aminoisobutyric acid (a nonmetabolizable aminoacid) and of 2-deoxyglucose was measured. NE did not inhibit TSH stimulation of the uptake of the two compounds. NE inhibited the action of dibutyryl cAMP (dbcAMP) on iodide uptake in a similar manner to TSH, but did not alter the cyclic adenosine monophosphate (cAMP) levels increased by TSH. The effects of different adrenoreceptor agonists and antagonists demonstrated that norepinephrine acts through an alpha1-adrenergic receptor. PMID- 9349588 TI - Soluble intercellular adhesion molecule-1 and Graves' disease. PMID- 9349589 TI - Octreoscan in Graves' ophthalmopathy. PMID- 9349590 TI - Transiently decreased sialylation of thyrotropin in a patient with the euthyroid sick syndrome. PMID- 9349591 TI - Hepatic expression of CCAAT/enhancer binding protein alpha: hormonal and metabolic regulation in rats. AB - There is a significant body of evidence which suggests that the alpha-isoform of the CCAAT/enhancer binding protein (C/EBP alpha) plays a central regulatory role in energy metabolism in the liver. However, there is little information available regarding regulation of its expression in this tissue. In this study, we examined the effect of hormones and diabetes on its expression in rat H4IIE hepatoma cells and in rat liver. Treatment of H4IIE cells with dexamethasone led to a threefold increase in C/EBP alpha mRNA within 4 h. Insulin treatment produced a bi-phasic response, initially reducing mRNA levels up to the 4 h time point, but after 8 h a twofold increase in C/EBP alpha mRNA was observed. Treatment with 8 chlorophenylthio-cAMP produced a twofold induction of C/EBP alpha mRNA after 8 h. Western analysis indicated that the changes in mRNA in response to hormonal treatment generally resulted in corresponding alterations in C/EBP alpha protein levels. Finally, we observed an inhibition of C/EBP alpha gene expression in streptozotocin-diabetic rat liver, reflected by a decrease in both mRNA and protein levels that were partially reversed by insulin treatment. These results indicate that the expression of C/EBP alpha in liver is under complex control by both hormonal and metabolic signals, which is consistent with its role as a trans -regulator of genes which play a role in energy metabolism. PMID- 9349592 TI - The effect of portal and peripheral insulin delivery on carbohydrate and lipid metabolism in a miniature pig model of human IDDM. AB - A pig model of insulin-dependent diabetes was used to examine the importance of the portal-systemic insulin gradient for whole-body metabolic control. Six pigs had jugular vein, portal vein, and carotid artery cannulae implanted before being made diabetic (150 mg kg[-1] streptozotocin). Each animal received 4 weeks of portal and 4 weeks of peripheral insulin delivery in random order. The blood glucose target range was 5-10 mmol x l(-1), and serum fructosamine and fasting and postprandial blood glucose concentrations were not different between peripheral and portal insulin infusion. Insulin requirement was not different between the 4 week infusion periods, but fasting peripheral insulin levels after peripheral delivery (124 +/- 16 (mean +/- SEM) pmol x l[-1]) were significantly higher (p < 0.05) than in portally infused (73.8 +/- 5.4 pmol x l[-1]) or pre diabetic control animals (68.4 +/- 3.6 pmol x l[-1]). Basal hepatic glucose output was also higher (p < 0.05) in peripherally (4.2 +/- 0.4 mg x kg[-1] x min[ 1]) than in portally infused animals (2.9 +/- 0.4 mg x kg[-1] x min[-1]) or controls (3.0 +/- 0.3 mg x kg[-1] x min[-1]). Clamp glucose metabolic clearance rate was, however, not different between the peripheral and portal insulin delivery routes (8.1 +/- 1.0 vs 9.0 +/- 0.7 ml x kg[-1] x min[-1]), although both were significantly lower (p < 0.05) than that measured in prediabetic control animals (11.7 +/- 1.0 ml x kg[-1] x min[-1]). Lipid profiles and subfractions were similar in all three groups. It is concluded that the portal route of delivery is superior to the peripheral in maintaining more appropriate insulin concentrations and control of hepatic glucose output, although in the absence of euglycaemia it is still associated with significant metabolic abnormalities. PMID- 9349593 TI - A 973 valine to methionine mutation of the human insulin receptor: interaction with insulin-receptor substrate-1 and Shc in HEK 293 cells. AB - A population-based study in the Netherlands has recently demonstrated that a mutation of the human insulin receptor (HIR-973 valine to methionine) is associated with hyperglycaemia and an increased prevalence of non-insulin dependent diabetes mellitus (NIDDM). The aim of the present study was to assess whether this mutation leads to a functional alteration of the insulin receptor. We prepared the HIR-973 mutant by in vitro mutagenesis. This mutant was transiently overexpressed in HEK 293 cells either alone or together with insulin receptor substrate-1 (IRS-1) or Shc. Insulin stimulated autophosphorylation, phosphorylation of the substrates IRS-1 and Shc as well as activation of phosphatidylinositol-3 (PI3)-kinase were studied. Autophosphorylation of HIR-973 and its susceptibility to hyperglycaemia induced inhibition was not different from HIR-wt. Human insulin receptor with a juxtamembrane deletion HIR-deltaJM which is known to impair HIR/IRS-1 interaction was used as control. While the HIR deltaJM induces a reduced IRS-1 phosphorylation HIR-973 showed even a slightly increased ability to phosphorylate IRS-1 (n = 7, 115% of control, p < 0.01). Shc phosphorylation was only mediated by HIR-wt and HIR-973 but not by HIR-deltaJM. Again a tendency to higher phosphorylation of Shc was seen with HIR-973 (n = 7, 109% of control, NS). When PI3-kinase activity was measured in IRS-1 precipitates similar activity was found for HIR-wt and HIR-973 whereas PI3-kinase stimulation was reduced with HIR-deltaJM. In summary, the data suggest that HIR-973 does not impair the first steps of the insulin signalling cascade. It is therefore unlikely that this mutation may cause cellular insulin resistance. The close vicinity of this mutation to insulin receptor domains which are involved in IRS-1 and Shc binding may, however, alter the interaction of the insulin receptor with these substrates. This could explain the slightly increased insulin effect on tyrosine phosphorylation of these docking proteins. These characteristics of HIR 973 might have a compensatory function of impaired signal transduction further downstream of the signalling chain in this specific subgroup of NIDDM patients. PMID- 9349594 TI - Relative contributions of advanced glycation and nitric oxide synthase inhibition to aminoguanidine-mediated renoprotection in diabetic rats. AB - Advanced glycation end products (AGEs) have previously been shown to be increased in the diabetic kidney. Aminoguanidine, an inhibitor of advanced glycation, has been shown to attenuate the development of AGEs as well as the progression of renal disease in experimental diabetes. However, the precise mechanisms through which aminoguanidine acts remain to be elucidated since it is also able to act as an inhibitor of nitric oxide synthase (NOS). This study has therefore compared the effects of aminoguanidine with the effects of two other inhibitors of NOS, L NAME and methylguanidine, on the development of experimental diabetic nephropathy. Diabetic rats were randomised to receive no treatment, aminoguanidine (1 g/l in drinking water), L-NAME (5 mg/l in drinking water) or methylguanidine (1 g/l in drinking water). Diabetic rats had increased levels of albuminuria and urinary nitrite/nitrate excretion when compared to control rats. Renal AGEs measured by fluorescence as well as by a carboxymethyllysine reactive radioimmunoassay, were elevated in diabetic rats. No changes in inducible NOS (iNOS) protein expression were detected in experimental diabetes nor did aminoguanidine affect iNOS expression. Aminoguanidine did not affect blood glucose or HbA1c but it did prevent increases in albuminuria, urinary nitrites/nitrates and renal AGE levels as measured by fluorescence and radioimmunoassay. L-NAME and methylguanidine did not retard the development of albuminuria, nor did they prevent increases in renal AGE levels, as assessed by fluorescence. However, these treatments did prevent increases in AGEs, as measured by radioimmunoassay. This study indicates that the renoprotective effect of aminoguanidine in experimental diabetes cannot be reproduced by L-NAME or methylguanidine. It is likely that the effect of aminoguanidine is mediated predominantly by decreased AGE formation rather than via NOS inhibition. It also raises the possibility that inhibition of fluorescent AGE formation may be more renoprotective than inhibition of the formation of carboxymethyllysine-containing AGEs. PMID- 9349595 TI - Length of metabolic normalization after rat islet cell transplantation depends on endocrine cell composition of graft and on donor age. AB - In vitro studies have demonstrated that beta-cell functions are negatively influenced by age and positively by the presence of glucagon producing alpha cells. This study examines whether the function of beta-cell grafts varies with the age of the donor and with the presence of other endocrine islet cells. Islet beta and endocrine non-beta-cells were purified from 10- to 30-week-old Lewis rats, and reaggregated into pure beta and mixed endocrine cell aggregates. Grafts consisted of 1.2 to 1.7 million beta cells with or without 0.6-0.7 million alpha cells. Their intraportal transplantation in 10-week-old streptozotocin-diabetic rats corrected hyperglycaemia in all experimental groups, with normal glucose tolerance curves at post-transplantation week (PT wk) 4. Recipients of mixed endocrine cell grafts from 10-week-old donors maintained a glucose tolerant state until PT wk 20, but turned glucose intolerant thereafter; only 1 of 12 animals was overtly diabetic at PT wk 64. Recipients of pure beta-cell grafts from 10 week-old donors became glucose-intolerant from PT wk 4 on, with 5 of 11 cases developing overt diabetes before PT wk 64. When grafts were prepared from 30-week old donors, metabolic deterioration started earlier, again with a more rapid loss for pure beta-cell grafts; at PT wk 64, virtually all recipients were overtly diabetic. It is concluded that delayed graft failure can be the consequence of an insufficient number of islet endocrine non-beta-cells as well as of a higher donor age. This observation can explain late failures in animal and human islet transplantation using marginally low beta-cell numbers. The interpretation of long-term studies on islet cell transplantation can benefit from the use of standardized grafts with well defined cellular composition. PMID- 9349596 TI - Contribution of proteolysis and de novo synthesis to alanine production in diabetic rat skeletal muscle: a 15N/1H nuclear magnetic resonance study. AB - To assess the role of leucine as a precursor of alanine alpha-amino nitrogen in skeletal muscle during diabetes, extensor digitorum longus muscles from control (n = 7 experiments) and streptozotocin-diabetic rats (n = 8 experiments) were isolated and superfused with [15N]leucine (3 mmol/l) in the presence of glucose (10 mmol/l) for 2 h. Muscle perchloric acid extraction was performed at the end of superfusion in order to quantify newly synthesized alanine by 15N/1H nuclear magnetic resonance. Release of [15N]alanine in the superfusion medium was also measured. The pool of newly synthesized [15N]alanine was significantly increased (approximately 40%) in extensor digitorum longus muscles from streptozotocin diabetic rats. Whereas a significant enhancement of total alanine release from muscle was induced by diabetes (20%), only a slight increase in [15N]alanine release was detectable under our experimental conditions. Consequently, we conclude that streptozotocin-diabetes in growing rats induces in skeletal muscle: 1) an increase in nitrogen exchange between leucine and alanine leading to newly synthesized [15N]alanine; and 2) an increase of total alanine release from muscle originating from both proteolysis and de novo synthesis. PMID- 9349597 TI - Soluble leucocyte adhesion molecules in diabetic retinopathy stimulate retinal capillary endothelial cell migration. AB - Diabetic retinal neovascularisation is considered to be a consequence of retinal ischaemia caused by capillary occlusion. Capillary occlusion is the result of microvascular thrombi in which erythrocytes, platelets and leucocytes each may play a role. We investigated the role of leucocytes in this process and the subsequent angiogenic response. We studied the serum levels of the soluble leucocyte adhesion molecules soluble E-Selectin, soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) in the serum of 93 patients with insulin-dependent diabetes (IDDM) and varying degrees of retinopathy and 47 healthy age and sex matched control subjects. We also measured the ability of serum to stimulate retinal capillary endothelial cell migration using an assay of angiogenesis in vitro. Soluble E-Selectin and sVCAM-1 levels were raised in all patients with IDDM (p < 0.001; p < 0.001) particularly those with retinopathy (p < 0.001; p < 0.001). Soluble E-Selectin levels were highest in the patients with severe non-proliferative diabetic retinopathy (p < 0.001) and sVCAM-1 levels were highest in patients with proliferative diabetic retinopathy (p < 0.01). In contrast soluble ICAM-1 levels were the same in patients and control subjects (p > 0.05). Soluble E-Selectin levels in diabetic patients were correlated with the level of glycated haemoglobin (p < 0.05). Retinal endothelial cell migration-inducing (ECMI) activity was increased in patients with IDDM (p < 0.01) in particular in those with retinopathy (p < 0.01). Furthermore, in vitro ECMI activity could be blocked by antibodies to sVCAM-1 and sE-Selectin. These data point to a functional role for leucocyte adhesion in the microvasculopathy of diabetic retinopathy and may have implications for the induction of retinal angiogenesis. PMID- 9349598 TI - Involvement of phosphoinositide 3-kinase in insulin stimulation of MAP-kinase and phosphorylation of protein kinase-B in human skeletal muscle: implications for glucose metabolism. AB - Isolated skeletal muscle from healthy individuals was used to evaluate the role of phosphoinositide 3-kinase (PI 3-kinase) in insulin signalling pathways regulating mitogen activated protein kinase (MAP-kinase) and protein kinase-B and to investigate whether MAP-kinase was involved in signalling pathways regulating glucose metabolism. Insulin stimulated glycogen synthase activity (approximately 1.7 fold), increased 3-o-methylglucose transport into human skeletal muscle strips (approximately 2 fold) and stimulated phosphorylation of the p42 ERK-2 isoform of MAP-kinase. This phosphorylation of p42 ERK2 was not blocked by the PI 3-kinase inhibitors LY294002 and wortmannin although it was blocked by the MAP kinase kinase (MEK) inhibitor PD 98059. However, PD98059 (up to 20 micromol/l) did not block insulin activation of glycogen synthase or stimulation of 3-o methylglucose transport. Wortmannin and LY294002 did block insulin stimulation of protein kinase-B (PKB) phosphorylation and stimulation of 3-o-methylglucose transport was inhibited by wortmannin (IC50 approximately 100 nmol/l). These results indicate that MAP-kinase is activated by insulin in human skeletal muscle by a PI 3-kinase independent pathway. Furthermore this activation is not necessary for insulin stimulation of glucose transport or activation of glycogen synthase in this tissue. PMID- 9349599 TI - Plasma leptin concentrations: gender differences and associations with metabolic risk factors for cardiovascular disease. AB - The cloning of the obese gene and the characterization of its protein product, leptin, has permitted the study of a new hormone potentially involved in the regulation of adipose tissue mass. The present study examined the gender differences in fasting plasma leptin concentration and its relationship to body fatness, adipose tissue distribution and the metabolic profile in samples of 91 men (mean age +/- SD: 37.3 +/- 4.8 years) and 48 women (38.5 +/- 6.8 years). Plasma leptin concentrations were strongly associated with body fat mass measured by underwater weighing [men: r = 0.80, p < 0.0001; women: r = 0.85, p < 0.0001]. In both genders, plasma leptin levels were also strongly correlated with waist girth as well as cross-sectional areas of abdominal subcutaneous and visceral adipose tissue measured by computed tomography. Women had, on average, plasma leptin concentrations that were three times higher than men. Furthermore, this gender difference remained significant when comparing men and women matched for similar levels of body fat mass. The associations between plasma leptin and lipoprotein concentrations were dependent of adiposity. In both men and women, elevated fasting plasma leptin levels were associated with higher plasma insulin concentrations, but only in women was the association maintained after correction for fat mass. Thus, results of the present study show that women have higher plasma leptin levels compared to men, independent of the concomitant variation in total body fat mass. Furthermore, our results also suggest that, in women, the association between plasma leptin and insulin concentrations is independent of adiposity, a finding which provides further support to the observation that adipose tissue leptin secretion may be upregulated by insulin. PMID- 9349600 TI - Multiple metabolic abnormalities in normal glucose tolerant relatives of NIDDM families. AB - Non-diabetic first degree relatives of non-insulin-dependent diabetic (NIDDM) families are at increased risk of developing diabetes mellitus, and have been studied to identify early metabolic abnormalities. Hormone concentrations measured by specific enzyme immunoassays were assessed in non-diabetic relatives of North European extraction, and control subjects with no family history of diabetes were matched for age, sex and ethnicity. A 75-g oral glucose tolerance test was conducted and those with newly diagnosed NIDDM were excluded. Basal insulin resistance was determined by homeostasis model assessment (HOMA), and hepatic insulin clearance by C-peptide:insulin molar ratio. Relatives (n = 150) were heavier (BMI: p < 0.0001) than the control subjects (n = 152), and had an increased prevalence of impaired glucose tolerance (15 vs 3%, p < 0.01). The relatives had increased fasting proinsulin levels and decreased C-peptide levels following the glucose load, while insulin levels were increased at all time points. To examine whether the differences in hormone levels were secondary to the differences in glucose tolerance and adiposity, we studied 100 normal glucose tolerant relatives and control subjects pair-matched for age, sex, waist-hip ratio and BMI. The differences in proinsulin levels were no longer apparent. However, the relatives remained more insulin resistant, and had decreased C peptide levels and C-peptide:insulin ratios at all time points. In conclusion, we have identified several metabolic abnormalities in the normal glucose tolerant relatives, and propose that the decreased hepatic insulin clearance helps to maintain normoglycaemia in the face of combined insulin resistance and decreased insulin secretion. PMID- 9349601 TI - High prevalence of risk factors for cardiovascular disease in parents of IDDM patients with albuminuria. AB - Life expectancy is shorter in the subset of insulin-dependent diabetic (IDDM) patients who are susceptible to kidney disease. Familial factors may be important. In this study the prevalence of cardiovascular disease mortality and morbidity and of risk factors for cardiovascular disease was compared in the parents of 31 IDDM patients with elevated albumin excretion rate (AER > 45 microg/min; group A) with that of parents of 31 insulin-dependent diabetic patients with normoalbuminuria (AER < 20 microg/min; group B). The two diabetic patient groups were matched for age and duration of disease. Information on deceased parents was obtained from death certificates and clinical records and morbidity for cardiovascular disease was ascertained using the World Health Organization questionnaire and Minnesota coded ECG. Hyperlipidaemia was defined as serum cholesterol higher than 6 mmol/l and/or plasma triglycerides higher than 2.3 mmol/l and/or lipid lowering therapy; arterial hypertension as systolic blood pressure higher than 140 mmHg and/or diastolic blood pressure higher than 90 mmHg and/or antihypertensive treatment. The percentage of dead parents was similar in the two groups (26 vs 20% for parents of group A vs group B, respectively), but the parents of the diabetic patients with elevated AER had died at a younger age (58 +/- 10 vs 70 +/- 14 years; p < 0.05). Parents of diabetic patients with nephropathy had a more than three times greater frequency of combined mortality and morbidity for cardiovascular disease than that of the parents of diabetic patients without nephropathy (26 vs 8%; odds ratio 3.96, 95% CI 1.3 to 12.2; p < 0.02). Living parents of group A had a higher prevalence of arterial hypertension (42 vs 14% p < 0.01) and hyperlipidaemia (49 vs 26% p < 0.05) as well as higher levels of lipoprotein (a) [median (range) 27.2 (1-107) vs 15.6 (0.2-98) mg/dl; p < 0.05]. They also had reduced insulin sensitivity [insulin tolerance test: median (range) K(itt) index: 3.7 (0.7-6.2) vs 4.8 (0.7-6.7)% per min; p < 0.05]. In the families of IDDM patients with elevated AER there was a higher frequency of risk factors for cardiovascular disease as well as a predisposition to cardiovascular disease events. This may help explain, in part, the high prevalence of cardiovascular disease mortality and morbidity in those IDDM patients who develop nephropathy. PMID- 9349602 TI - Peroxovanadate and insulin action in adipocytes from NIDDM patients. Evidence against a primary defect in tyrosine phosphorylation. AB - We studied the effects of insulin and the stable peroxovanadate compound potassium bisperoxopicolinatooxovanadate (bpV(pic)), a potent inhibitor of phosphotyrosine phosphatases, on lipolysis and glucose uptake in subcutaneous adipocytes from 10 male patients with non-insulin-dependent diabetes mellitus (NIDDM) and 10 matched non-diabetic control subjects. Lipolysis stimulated by isoprenaline or the cAMP analogue, 8-bromo-cyclic AMP (8-br-cAMP), was reduced by approximately 40% in NIDDM compared to control subjects. In both groups bpV(pic) exerted an antilipolytic effect that was similar to insulin (approximately 50 % inhibition). 14C-U-glucose uptake was dose-dependently increased by bpV(pic) treatment, but this effect and also that of insulin were impaired in NIDDM compared to control (bpV(pic) 1.6-fold vs 2.4-fold and insulin 2.2-fold vs 3.4 fold). Furthermore, low concentrations of bpV(pic) did not affect insulin stimulated glucose uptake, although tyrosine phosphorylation of the insulin receptor beta-subunit was clearly increased by bpV(pic). In conclusion, 1) beta adrenergic stimulation of lipolysis in vitro is attenuated in NIDDM adipocytes due to post-receptor mechanisms. 2) Both insulin and bpV(pic) decrease lipolysis and enhance glucose uptake in control as well as NIDDM adipocytes. The effect on glucose uptake, but not that on lipolysis, is impaired in NIDDM cells. 3) Peroxovanadate does not improve sensitivity and responsiveness to insulin in NIDDM adipocytes, showing that insulin-resistant glucose uptake in NIDDM is not overcome by phosphotyrosine-phosphatase inhibition and, thus, probably is not caused by impaired tyrosine phosphorylation events alone. PMID- 9349603 TI - Human leptin receptor gene in obese Japanese subjects: evidence against either obesity-causing mutations or association of sequence variants with obesity. AB - Leptin is an adipocyte-derived blood-borne satiety factor that acts on its cognate leptin receptor (Ob-R) in the hypothalamus, thereby regulating food intake and energy expenditure. To explore whether mutations in the Ob-R gene cause obesity in humans, we have searched for mutations in the gene for Ob-Rb, a biologically active receptor isoform, in obese Japanese subjects. We have also examined associations between such mutants and obesity in the Japanese. Genomic DNAs were used as templates in polymerase chain reaction (PCR) with primers selected to amplify exons 2 to 20 of the human Ob-Rb gene. Direct sequence analysis of the PCR products revealed 7 nucleotide sequence variants (Lys109Arg, Gln223Arg, Ser343Ser, Ser492Thr, Lys656Asn, Ala976Asp, and Pro1019Pro) in the Ob Rb coding region from 17 obese Japanese subjects with a family history of obesity (BMI 39.3 +/- 8.4 kg/m2). No missense and nonsense mutations were found such as those in Zucker fatty (fa/fa) rats and Koletsky (fa[k]/ fa[k]) rats. Nucleotide substitutions occurred at relatively high frequencies at codons 109, 223, 976, and 1019 (79, 91, 100, and 85%, respectively). Allele frequency of each variant determined by PCR-RFLP and PCR-single strand conformation polymorphism analyses showed no significant differences between 47 obese (BMI 35.1 +/- 6.5 kg/m2) and 68 non-obese (BMI 21.6 +/- 2.2 kg/m2) subjects. The present study represents the first report of sequence variants of the Ob-Rb gene in the Japanese and provides evidence against either obesity-causing mutations or association of sequence variants with obesity in obese Japanese subjects. PMID- 9349604 TI - Troglitazone increases the resistance of low density lipoprotein to oxidation in healthy volunteers. AB - The oxidative modification of low density lipoprotein is of importance in atherogenesis. Antioxidant supplementation has been shown, in published work, to increase low density lipoprotein resistance to oxidation in both healthy subjects and diabetic subjects; in animal studies a contemporary reduction in atherogenesis has been demonstrated. Troglitazone is a novel oral antidiabetic drug which has similarities in structure with vitamin E. The present study assessed the effect of troglitazone 400 mg twice daily for 2 weeks on the resistance of low density lipoprotein to oxidation in healthy male subjects. Ten subjects received troglitazone and ten received placebo in a randomised, placebo controlled, parallel-group design. The lag phase (a measure of the resistance of low density lipoprotein to oxidation) was determined by measurement of fluorescence development during copper-catalysed oxidative modification of low density lipoprotein. The lag phase was increased by 27 % (p < 0.001) at week 1 and by 24% (p < 0.001) at week 2 in the troglitazone treated group compared with the placebo group. A number of variables known to influence the resistance of low density lipoprotein to oxidation were measured. They included macronutrient consumption, plasma and lipoprotein lipid profile, alpha-tocopherol, beta carotene levels in low density lipoprotein, low density lipoprotein particle size, mono and polyunsaturated fatty acid content of low density lipoprotein and pre-formed low density lipoprotein hydroperoxide levels in low density lipoprotein. Troglitazone was associated with a significant reduction in the amount of pre-formed low density lipoprotein lipid hydroperoxides. At weeks 1 and 2, the low density lipoprotein hydroperoxide content was 17% (p < 0.05) and 18% (p < 0.05) lower in the troglitazone group compared to placebo, respectively. In summary the increase in lag phase duration in the troglitazone group appeared to be due to the compound's activity as an antioxidant and to its ability to reduce the amount of preformed low density lipoprotein lipid hydroperoxides. This antioxidant activity could provide considerable benefit to diabetic patients where atherosclerosis accounts for the majority of total mortality. PMID- 9349605 TI - Protein intake and urinary albumin excretion rates in the EURODIAB IDDM Complications Study. AB - For people with insulin-dependent diabetes mellitus (IDDM) renal disease represents a life-threatening and costly complication. The EURODIAB IDDM Complications Study, a cross-sectional, clinic-based study, was designed to determine the prevalence of renal complications and putative risk factors in stratified samples of European individuals with IDDM. The present study examined the relationship between dietary protein intake and urinary albumin excretion rate (AER). Food intake was assessed centrally by a standardized 3-day dietary record. Urinary AER was determined in a central laboratory from a timed 24-h urine collection. Complete data were available from 2696 persons with IDDM from 30 centres in 16 European countries. In individuals who reported protein consumption less than 20% of total food energy intake, mean AER was below 20 microg/min. In those in whom protein intake constituted more than 20%, mean AER increased, a trend particularly pronounced in individuals with hypertension and/or poor metabolic control. Trends reached statistical significance for intakes of total protein (% of energy, p = 0.01) and animal protein (% of energy, p = 0.02), while no association was seen for vegetable protein (p = 0.83). These findings support the current recommendation for people with diabetes not to exceed a protein intake of 20% of total energy. Monitoring and adjustment of dietary protein appears particularly desirable for individuals with AER exceeding 20 microg/min (approximately 30 mg/24h), especially when arterial pressure is raised and/or diabetic control is poor. PMID- 9349607 TI - In utero undernutrition impairs rat beta-cell development. AB - The role of nutrition on the development of the endocrine pancreas was studied in a rat model obtained by maternal food restriction. A 50% food restriction was applied to female rats from day 15 of pregnancy and resulted in intrauterine growth-retardation (IUGR) in the offspring. At day 1 postnatal, beta-cell mass was significantly decreased in IUGR pups as compared to controls (0.70 +/- 0.06 vs 1.07 +/- 0.06 mg, p < 0.0001), as well as insulin content. This change in beta cell mass can be attributed to a reduced number of islets, since the density of insulin-positive aggregates in pancreatic sections of IUGR rats was 20% lower than in controls. Proliferative capacity of beta cells, as measured by 5-bromo-2 deoxyuridine (BrdU) labelling index, was not altered in growth-retarded animals. Body as well as pancreatic weight were fully recovered in IUGR pups after 21 days of normal feeding by control mothers. However, these animals retained a 25% decrease in insulin content, 40% decrease in beta-cell mass (1.58 +/- 0.18 vs 2.78 +/- 0.42 mg, p < 0.001) and a strong reduction in the density of insulin positive aggregates per cm2, as compared to controls, suggesting that the total islet number was likely to be reduced. Beta-cell proliferative capacity remained normal. In conclusion, in utero undernutrition in rats does not impede postnatal growth but durably impairs beta-cell development. Impairment of beta-cell differentiation might be suggested. PMID- 9349608 TI - Could the aetiology of IDDM be multifactorial? PMID- 9349606 TI - Mutational analysis of the coding region of the uncoupling protein 2 gene in obese NIDDM patients: impact of a common amino acid polymorphism on juvenile and maturity onset forms of obesity and insulin resistance. AB - Recently, a gene encoding a novel human uncoupling protein, designated UCP2, was discovered. The murine UCP2 was mapped to a region on mouse chromosome 7 which in several models has been shown to be linked to obesity and hyperinsulinaemia. Single strand conformation polymorphism (SSCP) analysis and direct sequencing of the coding region of the UCP2 gene in 35 obese Caucasian NIDDM patients of Danish ancestry revealed one nucleotide substitution, replacing an alanine with a valine at codon 55. The amino acid polymorphism was present in 24 of the 35 (69%) examined subjects. The allelic frequency of the A/V55 variant was 48.3% (95% CI: 42.5-54.1%) among 144 subjects with juvenile onset obesity, 45.6% (40.5-50.7%) among 182 subjects randomly selected at the draft board examination, and 45.5% (37.1-53.9%) among lean control subjects selected from the same study cohort. Within these cohorts there were no differences in BMI values at different ages among wild-type carriers and A/V55 carriers. In a population-based sample of 369 young healthy Caucasians the variant showed no association with alterations in BMI, waist-to-hip ratio, fat mass or weight gain during childhood or adolescence. The A/V55 polymorphism was not related to alterations in fasting values of serum insulin and C-peptide or to an impaired insulin sensitivity index. We conclude that genetic variability in the human UCP2 gene is not a common factor contributing to NIDDM in obese Danish Caucasian subjects and the common A/V55 amino acid polymorphism of the gene is not implicated in the pathogenesis of juvenile or maturity onset obesity or insulin resistance in Caucasians. PMID- 9349609 TI - No association of ALDH2 genotype in MELAS. PMID- 9349610 TI - 1997 Minkowski Prize, 32nd Minkowski Lecture: Genetic approaches of NIDDM. PMID- 9349611 TI - 1997 Castelli Pedroli Prize, 12th Camillo Golgi Lecture. PMID- 9349612 TI - Filling in the blanks. PMID- 9349613 TI - HLA class I DNA typing in organ transplantation. PMID- 9349614 TI - The absence of DR51 in a DRB5-positive individual DR2ES is caused by a null allele (DRB5*0108N) AB - DR51, a protein encoded by the DRB5 gene, was shown to be present in almost all DR2-positive haplotypes. Exceptions were reported, some DR2-negative samples were shown to be DR51 positive and in a number of DR2-positive samples no DR51 antigen could be demonstrated. In some of them lack of the DRB5 gene was the cause of the absence of DR51 but in others the DRB5 gene was present without resulting in a detectable gene product. Many of these variants were studied in detail in previous international workshops. One of them was DR2ES from our laboratory. She is a DR15-positive DR51-negative individual of oriental origin with a clearly demonstrable DRB5*01 allele when typed by molecular techniques. To unravel the molecular mechanism responsible for the defect in expression, cDNA and DNA encoding the defective DRB5 allele were analyzed. Nucleotide sequence analysis of exon 2 showed no differences from the sequence of DRB5*0102. However, when exon 3 was examined a difference in length was noticed due to a deletion of 19 nucleotides between codon 161 and 168. The deletion caused a frameshift and a premature stopcodon resulting in a null allele. The same allele could be demonstrated in 6 other unrelated individuals of oriental origin as well as in 5 individuals from South Africa. The absence of the DR51 protein was explained by the presence of an alteration in the DRB5 allele resulting in a null allele. The allele has been officially named DRB5*0108N. This is the first description of a null allele of the DRB5 gene. PMID- 9349615 TI - Full-length cDNA nucleotide sequence of a serologically undetectable HLA-DQA1 allele: HLA-DQA1*"LA". AB - This study describes the characterization of a serological HLA-DQ"blank" specificity that segregates with the HLA-A2, -B7, -DR14, -DR52 haplotype. Although conventional serological typing techniques could not detect an HLA-DQ product on the haplotype positive for the HLA-DQ"blank" specificity, sequence specific oligonucleotide (SSO) dot-blot analysis demonstrated the presence of the HLA-DQA1*01 and HLA-DQB1*05 alleles. Full-length cDNA nucleotide sequence analysis revealed that the HLA-DQB1 allele that segregated with the HLA-DQ"blank" specificity was identical to HLA-DQB1*05031. As for the HLA DQA1 allele, one nucleotide substitution distinguished the HLA-DQA1 "blank" allele from HLA DQA1*0104. In exon 2 at nucleotide position 304 a C was substituted for a T (Arg- >Cys). Pending official recognition by the WHO Nomenclature Committee, this HLA DQA1 "blank" allele is termed HLA-DQA1*"LA". Furthermore, it is postulated that the introduction of cysteine at amino acid position 102 abrogates the classical HLA-DQ1 specificity. PMID- 9349616 TI - An intronic mutation responsible for a low level of expression of an HLA-A*24 allele. AB - HLA class I typing performed in parallel by molecular biology and serology has revealed cases where an HLA class I allele was identified but the corresponding antigen on the cell surface was not detected. In the present report, we describe three members of a family in whom an HLA-A24 allele identified at the molecular level was typed as A "blank" by lymphocytotoxicity. This serologically blank antigen was nevertheless faintly detectable by isoelectric focusing (IEF) and FACS analyses. Sequencing of the HLA-A*24 allele from the promoter region to the eighth exonic region revealed a point mutation in the acceptor site of the second intron as compared to the normal HLA-A*24 allele. This mutation could lead to incorrect processing of mRNA through a cryptic acceptor site located at the beginning of the third exon and hence to alternative splicing with a frame shift introducing an early stop codon into the fourth exon. PMID- 9349617 TI - A nucleotide insertion in exon 4 is responsible for the absence of expression of an HLA-A*01 allele. AB - HLA class I typing performed in parallel by molecular biology and serology has revealed cases where an HLA class I allele was identified whereas the corresponding antigen was not detected on the cell surface. In the present report, we describe four members of a family in whom an HLA-A1 allele identified at the molecular level was typed as A "blank" by lymphocytotoxicity. This serologically blank antigen was undetectable by isoelectric focusing (IEF). Sequencing of the HLA-A*01 allele from the promoter region to the eighth exonic region revealed insertion of a "C" nucleotide at the beginning of the fourth exon as compared to the common HLA-A*0101 allele. This mutation causes a frame shift, giving rise to an early stop codon in the fourth exon. PMID- 9349618 TI - An HLA-B null allele (B*1526N) with a stop codon in exon 3 generated by a point mutation. AB - An HLA-B null allele was identified in a Japanese family during histocompatibility testing for bone marrow transplantation. The propositus was a healthy Japanese woman with three children, and her parents were cousins. Serological HLA typing of the family members indicated that the propositus was homozygous for the A24-Cw4-B blank (B null)-DR4.2-DQ3 haplotype. Total RNA was extracted from peripheral blood of the propositus was converted to first-strand cDNA using reverse transcriptase. The cDNA was amplified by the polymerase chain reaction (PCR) using HLA-B locus-specific primers. The PCR product showed no change in size upon polyacrylamide gel electrophoresis (PAGE) compared to that of normal controls, suggesting that HLA-B gene mRNA was normally expressed. The nucleotide sequence of the cDNA was the same as that of B*1501 except at nucleotide 369 or codon 123, where C was replaced with A; TAC encodes Tyr whereas TAA is a stop codon. This point mutation may have truncated the HLA-B molecule of the propositus, resulting in the negative results we obtained with anti-HLA-B sera. PMID- 9349619 TI - HLA class I DNA typing of 215 "HLA-A, -B, -DR zero mismatched" kidney transplants. AB - DNA typing for HLA class II improves the typing quality and this was shown previously to be relevant for kidney graft survival. In this project we addressed the question whether molecular typing for HLA class I also increases the efficacy of HLA matching in kidney transplantation. 215 HLA-A,-B,-DR zero-mismatched donor/recipient pairs as defined by serological typing were selected. Retrospective HLA-A and HLA-B typing was performed both by the PCR-SSP and the PCR-SSOP method. DNA typing for HLA-A revealed discrepant results to serology in 5.7% of the donors and 2.8% of the recipients. HLA-B typing discrepancies were found in 6.6% of the donors and 5.6% of the recipients. 10.4% of the donors and 6.5% of the recipients showed either an HLA-A or an HLA-B discrepancy Nearly one third of the HLA-A discrepancies affected A19 splits. The most common reason for HLA-A discrepancies was the erroneous assignment of serological blanks, whereas HLA-B errors were caused mainly by the assignment of incorrect specificities. DNA typing allowed the definition of HLA-A and -B split specificities in all 118 "splitable" cases for which only broad specificities were reported based on serological typing. A total of 183 DNA class I compatible transplants had a 15% higher one-year graft survival rate than 32 transplants for which DNA typing revealed a class I incompatibility PMID- 9349620 TI - A 25% error rate in serologic typing of HLA-B homozygotes. AB - The microlymphocytotoxicity technique has been the accepted method for HLA class I typing since the early 1960s. However, it is often difficult to distinguish two related alleles expressed in an individual due to the cross-reactive nature of the alloantibodies used in this technique. This is especially evident at the HLA B locus, whose more than 180 alleles fall into only 4 major interrelated cross reactive antigen groups. To estimate the error rate in serologic typing due to the cross-reactive nature of sera, we used polymerase chain reaction with sequence-specific primers (PCR-SSP) amplification to retype 40 individuals who were previously typed as serologic HLA-B locus homozygotes. PCR-SSP revealed that 10 of these 40 individuals (25%) were actually heterozygous at their HLA-B loci. The HLA-B locus alleles of 9 of these 10 discrepant individuals were further analyzed by denaturing gradient gel electrophoresis followed by direct sequencing. The sequence analysis confirmed that all nine individuals were indeed HLA-B locus heterozygotes. This surprisingly high error rate in serologic definition of HLA-B molecules argues for the use of rapid DNA-based techniques in HLA class I typing, even in the setting of solid organ transplantation. PMID- 9349621 TI - Analysis of HLA-A and -B serologic typing of bone marrow registry donors using polymerase chain reaction with sequence-specific oligonucleotide probes and DNA sequencing. AB - Unrelated volunteer donors (69) recruited by the National Marrow Donor Program were HLA typed by DNA-based methods for both the HLA-A and -B loci. Each donor had been previously typed by serology by at least two independent laboratories. Of the 69 samples, all serologic laboratories were in concordance for HLA-A in 62 typed samples and for HLA-B in 48 typed samples. Of the serologically concordant samples, 5 samples typed for HLA-A and 7 samples typed for HLA-B received DNA and serology types differing in their level of resolution. One sample typed for HLA-A and 3 samples typed for HLA-B by DNA methods gave different results from their serologic assignments. Of the samples exhibiting disparities among the different serologic typing laboratories, the DNA-defined types of 7 samples typed for HLA-A and 18 samples typed for HLA-B were consistent with at least one of the serologic assignments. The DNA types for the remaining 3 HLA-B typed samples did not agree with the serologic assignments and their alleles were subsequently sequenced. One of these sequences was a previously undefined allele, B*1537. Sharing of polymorphic sequences among HLA allelic products creates difficulties for consistent serologic assignments of some types complicating the process of identifying potential donors from bone marrow registries. Thus, the use of DNA based typing techniques for characterization of donor class I types should allow a more consistent definition of types and should speed the donor selection process. PMID- 9349622 TI - HLA-A typing: comparison between serology, the amplification refractory mutation system with polymerase chain reaction and sequencing. AB - In this study we typed HLA-A polymorphisms by a new sequence-based typing (SBT) method, which involved one PCR reaction and four sequencing reactions covering exon 2 and exon 3. This method allowed complete identification of all known HLA-A alleles and revealed the presence of a new allele, named HLA-A*2608. We also introduced sequencing of exon 4 for some samples in order to discriminate the allelic pairs that are identical in exon 2 and 3, thus improving SBT resolution. Finally, we compared the results obtained by SBT with data obtained by serological typing and the amplification refractory mutation system (ARMS-PCR). Together, our results suggest that the SBT here described provides an optimal HLA A typing technique that may be useful in selecting donor-recipient pairs in bone marrow transplantation between unrelated individuals. PMID- 9349623 TI - Accurate typing of HLA-A antigens and analysis of serological deficiencies. AB - We are reporting the results of HLA-A typing by PCR-SSOP complemented by PCR-SSP of samples obtained from the National Marrow Donor Program (NMDP). These samples were a representative group from 2486 tested in duplicate by serology. A total of 390 samples gave HLA-A discrepant results. Comparing the molecular typing results of 238 samples (samples with available DNA) with the serological typing results, 54 homozygotes and 184 heterozygotes produced a total of 422 assignments by molecular methods. We found assignment discrepancies in 147/422 (35%) in laboratory 1 and 144/422 (34%) in laboratory 2 (a combined group of 4 NMDP laboratories; laboratory 1 is not included). The serological discrepancies found were of 3 categories: a) false negatives, b) incomplete typing (discrepancies due to the level of resolution within a cross-reactive or CREG group) and c) false positives. Major problems were identified using serology for typing HLA-A antigens: a) inability to identify all WHO-recognized specificities, more frequently in non-Caucasians or in HLA-A specificities known to be found more frequently in non-Caucasians for laboratory 1 and incorrect assignments of A19 specificities in laboratory 2, b) incorrect assignments in cells with poor viability and c) false-positive assignments in homozygotes. We propose a possible strategy to type HLA-A specificities with two steps: a) a minimum of serology for typing specificities for common CREG groups: A1, A2, A3, A11, A9, A10, A28, A19. However, a given laboratory can determine the level of serological assignments needed as a first step. And b) molecular methods to identify splits: A23, A24, A29, A30, A31, A32, A33, A34, A36, A66, A74 and A80. The technique described is useful for large-scale bone marrow donor typings for cells with poor viability, and for resolving ambiguous results including false-positive assignments of homozygous cells. PMID- 9349625 TI - Comparison of typing results by serology and polymerase chain reaction with sequence-specific primers for HLA-Cw in 650 individuals. AB - HLA-Cw typing by standard serological techniques is associated with a high frequency of blanks, and reliable typing reagents for several of the Cw specificities are scarce. We evaluated the PCR-SSP technique for Cw typing in 370 kidney transplant patients and 280 healthy blood donors. Serological typing of all individuals was performed in our laboratory from 1995 to 1997 using commercially available tissue-typing trays. Comparison between serological and PCR-SSP typing revealed a discrepancy rate of 33.6% (n= 94) in blood donors and 32.4%) (n=120) in kidney recipients. Incorrect antigen assignments occurred only rarely (3.6% of the blood donors and 3.2% of the kidney recipients). The vast majority of discrepancies were due to antigens that were not detected serologically. In 26 individuals no Cw antigen was detected by serological typing, whereas PCR-SSP showed 1 allele in 13 and 2 alleles in the other 13 cases. Another 269 individuals were typed serologically with one blank (presumably homozygous). Of these, only 108 were confirmed to be homozygous, whereas an additional Cw allele was found in the remaining 161 cases using the SSP technique. Most of the "missed" specificities (86.5%) were those for which serological reagents were not available (HLA-Cw*12-*17). The most commonly "missed" specificity was HLA-Cw*1203, which occurred in 13.9% of the healthy blood donors. These results indicate that serological HLA-Cw typing is insufficient for examining the clinical importance of HLA-Cw matching in transplantation. Future studies based on molecular typing should allow the proper investigation of HLA-Cw matching in kidney and bone marrow transplantation. PMID- 9349624 TI - Error rate for HLA-B antigen assignment by serology: implications for proficiency testing and utilization of DNA-based typing methods. AB - Until recently, the majority of HLA class I typing has been performed by serology. Expensive commercial typing trays are frequently used for testing non Caucasian subjects and new strategies using DNA-based methods have been adopted for improving clinical histocompatibility testing results and adapted as supplements in proficiency testing. A double-blind comparison of the typing of HLA-B specificities in 40 samples was carried out between serology and two polymerase chain reaction (PCR) methods, PCR amplification with sequence-specific primers (PCR-SSP) and PCR amplification and subsequent hybridization with sequence-specific oligonucleotide probes (PCR-SSOP). The results demonstrated 22.5% misassignments of HLA-B antigens by serology. There was complete concordance between the results obtained with the two PCR based typing methods. A second panel of 20 donor samples with incomplete or ambiguous serologic results was analyzed by PCR-SSP and SSOP Both PCR methods identified correctly the HLA-B antigens. Our results suggest that more accurate typing results can be achieved by complementing serologic testing with DNA-based typing techniques. The level of resolution for HLA-B antigen assignment can be obtained by this combination of serology and limited DNA-based typing is equivalent to the HLA-B specificities defined by the WHO-HLA Committee. This level of resolution cannot routinely be achieved in clinical histocompatibility testing or in proficiency testing using serologic reagents only. PMID- 9349626 TI - The search for HLA-matched donors: a summary of HLA-A*, -B*, -DRB1/3/4/5* alleles and their association with serologically defined HLA-A, -B, -DR antigens. AB - This report summarizes data obtained from several large studies including the WHO HLA Nomenclature Committee, the International Cell Exchange, UCLA, the British Society for Histocompatibility and Immunogenetics Rare Cell Exchange and the National Marrow Donor Program and individual laboratories aimed at identifying a serologic type for specific HLA-A,-B,-DRB allelic products. Alleles that are poorly characterized at the serologic level are indicated and an approach is suggested for obtaining the information needed to clarify their serologic typing. The tables provided will be useful in guiding searches for an unrelated donor in which patient and/or potential donors are typed either by serology or by DNA based methods and will provide a "dictionary" of potential equivalents between HLA "types" obtained by the two methods. PMID- 9349627 TI - Nomenclature for factors of the HLA system, update May/June 1997. WHO Nomenclature Committee for Factors of the HLA System. PMID- 9349628 TI - Inhibition of CREB- and cAMP response element-mediated gene transcription by the immunosuppressive drugs cyclosporin A and FK506 in T cells. AB - The clinically important immunosuppressant drugs cyclosporin A and FK506 (tacrolimus) inhibit in T-cells calcineurin phosphatase activity and nuclear translocation of the cytosolic component of the transcription factor nuclear factor of activated T-cells (NF-ATc) that is involved in the induction of early genes during T-cell activation. This effect has been proposed to explain at least part of the immunosuppressive effect of these drugs. Previous studies in pancreatic islet cell lines have shown that cyclosporin A and FK506 through inhibition of calcineurin interfere also with the function of the transcription factor cAMP response element binding protein (CREB) that is activated by cAMP and calcium signals and binds to cAMP/calcium response elements (CRE). By transient expression of CRE-reporter genes or GAL4-CREB fusion proteins, the present study shows that inhibition of CREB/CRE-directed transcription by cyclosporin A and FK506 occurs in a great variety of cell types including in cell lines derived from tissues in which adverse effects of the immunosuppressants develop. CREB activity and CRE-mediated transcription was blocked by these drugs also in Jurkat T-cells. When taken together with recent evidence for an essential role of CREB in T-cell activation and proliferation, the present results suggest that inhibition of CREB/CRE-directed transcription may be a molecular mechanism of the immunosuppressive effect of cyclosporin A and FK506. PMID- 9349629 TI - A test system for leukotriene synthesis inhibitors based on the in-vitro differentiation of the human leukemic cell lines HL-60 and Mono Mac 6. AB - Differentiation of HL-60 cells along the granulocytic lineage by DMSO in the presence of transforming growth factor-beta and low concentrations of 1,25 dihydroxyvitamin D3 leads to the upregulation of 5-lipoxygenase activity in 100,000 g supernatants and intact cells to levels which are comparable to normal granulocytes. Similarly, differentiation of the human monocytic cell line Mono Mac 6 by 1,25-dihydroxyvitamin D3 and transforming growth factor-beta strongly upregulates the 5-lipoxygenase pathway. Here, we describe an assay system for leukotriene biosynthesis inhibitors which is based on the in-vitro differentiation of HL-60 and Mono Mac 6 cells. Different leukotriene biosynthesis inhibitors like the nonredox type inhibitor ZM 230487, the redox type inhibitor BW A4C and the FLAP inhibitor MK886 were tested and the results were compared with an assay system based on normal human granulocytes. ZM 230487, BWA4C and MK886 showed similar potencies in these cell lines as compared to normal leukocytes. Thus, the in-vitro differentiation of HL-60 and Mono Mac 6 cells provides an excellent model for the screening of drugs affecting the 5 lipoxygenase pathway. PMID- 9349630 TI - Selective visualization of rat brain 5-HT2A receptors by autoradiography with [3H]MDL 100,907. AB - The recently developed 5-HT2A receptor selective antagonist [3H]MDL100,907 ((+/ )2,3-dimethoxyphenyl-1-[2-(4-piperidine)-methanol]) has been characterized as a radioligand for the autoradiographic visualization of these receptors. [3H]MDL100,907 binding to rat brain tissue sections was saturable, had sub nanomolar affinity (Kd = 0.2-0.3 nM), and presented a pharmacological profile consistent with its binding to 5-HT2A receptors (rank order of affinity for [3H]MDL100,907-labelled receptors: MDL100,907 > spiperone > ketanserin > mesulergine). The distribution of receptors labelled by [3H]MDL100,907 was compared to the autoradiographical patterns obtained with [3H]Ketanserin, [3H]Mesulergine, and [3H]RP62203 (N-[3-[4-(4-fluorophenyl)piperazin-1-y1]propyl] 1,8-naphtalenes ultam) and to the distribution of 5-HT2A receptor mRNA as determined by in situ hybridization. As opposed to the other radioligands, [3H]MDL100,907 labelled a single population of sites (5-HT2A receptors) and presented extremely low levels of non-specific binding. The close similarity of the distributions of [3H]MDL100,907-labelled receptors and 5-HT2A mRNA further supports the selectivity of this radioligand for 5-HT2A receptors and suggests a predominant somatodendritic localization of these receptors. The present results point to [3H]MDL100,907 as the ligand of choice for the autoradiographic visualization of 5-HT2A receptors. PMID- 9349631 TI - Release of [3H]dopamine from guinea pig striatal slices is modulated by sigma1 receptor agonists. AB - Sigma receptors are found in motor and limbic areas in the brains of humans, non human primates, and rodents. The most extensive pharmacological studies of ligand binding to sigma receptors have utilized brain tissue from guinea pigs, where two subtypes of sigma receptor, designated sigma1 and sigma2, have been identified. Few functional roles for sigma receptors have been described. Their location in guinea pig striatum, a terminal field of dopaminergic projections arising from the substantia nigra, suggested that this tissue would be a logical choice in which to examine physiological properties of sigma receptor activation. We found that sigma1 receptor agonists inhibited N-methyl-D-aspartate-stimulated [3H]dopamine release from guinea pig striatal slices in a concentration-dependent manner. The inhibition by sigma1 receptor agonists was reversed by a selective sigma1 receptor antagonist, as well as by a non-subtype-selective sigma receptor antagonist. The ability of agonists working through sigma1 receptors, but not through sigma2 receptors, to inhibit the stimulated release of catecholamines appears to be a unique characteristic of guinea pig striatum. We have previously reported that in rat striatum and hippocampus, as well as in guinea pig nucleus accumbens, prefrontal cortex, and hippocampus, activation of either sigma receptor subtype inhibits such release. Stimulated release of [3H]dopamine from guinea pig striatum was also inhibited by the phencyclidine receptor agonist dizocilpine, but this inhibition was not reversed by the sigma receptor antagonists. Therefore, the inhibition produced by sigma receptor agonists was not mediated via the phencyclidine binding site within the N-methyl-D-aspartate operated cation channel. Our findings support the hypothesis that sigma receptor activation provides a mechanism of modulating dopamine release from striatum, and that striatal tissue from guinea pigs appears to be an appropriate model for characterizing sigma1 receptor-mediated effects. PMID- 9349632 TI - Evidence for partial agonist properties of daltroban (BM 13,505) at TP receptors in the anaesthetized open-chest rat. AB - We sought to determine whether the intrinsic pulmonary hypertensive activity of the purported thromboxane A2/prostanoid (TP) receptor antagonist, daltroban, was mediated by TP receptors, using the high efficacy TP receptor agonist, U-46619, and the silent TP receptor antagonist, SQ 29,548. In pentobarbitone-anesthetized, open-chest rats (n = 4-10 per group), non-cumulative injections of U-46619, dose dependently increased mean pulmonary arterial pressure (MPAP) with an ED50 (geometric mean with 95% confidence limits in parentheses) of 1.4 (1.1-2.3) microg/kg i.v.. Daltroban increased MPAP in a bell-shaped manner, with an apparent ED50 [29 (21-35) microg/kg i.v.] being 21 fold less potent than that of U-46619. The maximal pulmonary hypertensive responses evoked by daltroban represented about half those induced by U-46619 (25.4+/-1.0 vs. 12.7+/-2 mmHg; P < 0.05 between groups). The TP receptor antagonist SQ 29,548 fully antagonized increases in MPAP evoked by equihypertensive doses of U-46619 (1.25 microg/kg) or daltroban (80 microg/kg). Further experiments were carried out to determine whether daltroban antagonized the pulmonary hypertensive responses evoked by the high efficacy agonist, U-46619, or by itself as receptor theory would predict for a partial agonist. Daltroban (10-2500 microg/kg) antagonized, although not fully, U-46619 (20 microg/kg)-evoked pulmonary hypertensive responses, since prominent intrinsic pulmonary hypertensive effects of daltroban were observed in the same range of doses. Furthermore, in contrast to U-46619 (1.25 microg/kg), daltroban (80 microg/kg) failed to evoke a second pulmonary hypertensive response following a previous injection, as would be expected for a partial agonist. Collectively, the results strongly suggest that daltroban behaves as a partial agonist at TP receptors in the pulmonary vascular bed of the rat in vivo. PMID- 9349633 TI - Interaction between thiol-modifying agents and P1075, a K(ATP) channel opener, in rat isolated aorta. AB - In vascular smooth muscle, openers of ATP-dependent potassium channels (K(ATP) channels), such as P1075 (N-cyano-N'-(1,1-dimethylpropyl)-N"-3-pyridylguanidine), produce relaxation. In this study we have investigated the effects of thiol modifying agents on the binding of P1075 and on the 86Rb+ efflux stimulating and vasorelaxant effects of the opener in rat aortic rings. The increase in 86Rb+ efflux induced by P1075 was taken as a qualitative measure of K+ channel opening. The hydrophilic SH-group-oxidizing substance, thimerosal (1 to 100 microM), abolished specific binding of [3H]-P1075 with an IC50 value of 7.6+/-1.2 microM; at 30 microM, the half time for inhibition was 38 min. Two other thioloxidizing agents, PMB (4-hydroxy-mercuribenzoic acid) and DTBNP (2,2'-dithio-bis(5 nitropyridine)), inhibited binding up to 86% and 44%, respectively. The disulphide bond reducing substance, DTT (1,4-dithiothreitol, 0.1 to 1 mM), reduced [3H]-P1075 binding by up to 20% and partially reversed the inhibitory effect of thimerosal. In 86Rb+ efflux experiments, thimerosal (3 to 100 microM) concentration-dependently increased basal efflux but inhibited P1075-stimulated tracer efflux with an IC50 value of 7+/-1 microM. The inhibitory effect occurred with a half-time of approximately 8 min and was essentially reversed by DTT. In rings precontracted with noradrenaline, thimerosal inhibited the vasorelaxant effect in a noncompetitive manner, shifting the concentration-relaxation curves to the right and reducing maximum relaxation. The data show that oxidation of thiol groups interferes with the binding of the K(ATP) channel opener, P1075; concomitantly, the 86Rb+ efflux stimulating and the vasorelaxant effects are inhibited. Reduction of disulphide bonds by DTT has only minor effects on the action of P1075. Collectively, the results suggest that intact thiol groups are essential for the functioning of the K(ATP) channel in rat aorta. The different kinetics governing the inhibition of opener binding and of opener-stimulated 86Rb+ efflux suggest that the SH-groups involved in the two processes differ in their accessibility to thimerosal and/or in their reactivity. PMID- 9349634 TI - Role of nitric oxide, prostaglandins and tyrosine kinase in vascular endothelial growth factor-induced increase in vascular permeability in mouse skin. AB - We investigated role of nitric oxide (NO), prostaglandins (PG) and tyrosine kinase in vascular endothelial growth factor (VEGF)-induced increase in vascular permeability in mouse skin. Subcutaneous injection of VEGF (0.5-2.0 ng/site) induced dose- and time-dependent increase in vascular permeability at the injection site determined by a leakage of Pontamine sky blue. VEGF (1 ng/site) induced dye leakage was partially inhibited by N(G)-nitro-L-arginine methyl ester (an inhibitor for both constitutive and inducible NO synthase) (5 and 10 mg/kg, i.v.) and by aminoguanidine (a selective inducible NO synthase inhibitor) (5-20 mg/kg, i.v.), but not by an inactive enantiomer, N(G)-nitro-D-arginine methyl ester (10 mg/kg, i.v.). Pretreatment with an intraperitoneal injection of indomethacin (a nonselective cyclooxygenase inhibitor) (5 mg/kg) or N-(2 cyclohexyloxy-4-nitrophenyl) methanesulphonamide (a cyclooxygenase-2 selective inhibitor) (1-100 microg/kg) almost completely inhibited the effect of VEGF (1 ng/site). Coadministration of PGE2 (3 and 30 nmol/site) with VEGF did not restore the inhibitory effect of indomethacin on VEGF (1 ng/site)-induced increase in vascular permeability. Lavendustin A (a selective tyrosine kinase inhibitor) (10 and 50 microg/kg, s.c.) dose-relatedly inhibited the VEGF (1 ng/site)-induced increase in dye leakage, whereas its negative control, lavendustin B (10 microg/kg, s.c.) had no effect. Another tyrosine kinase inhibitor, genistein (2.5 mg/kg, s.c.) also inhibited the response. Cycloheximide (a protein biosynthesis inhibitor) (35 mg/kg, s.c.) suppressed the response of VEGF (1 ng/site). Histologically, no cellular infiltration was observed in the area of VEGF injection. These results suggest that increased vascular permeability induced by VEGF is mediated by local production of NO and arachidonic acid metabolites other than PGE2, which are most probably produced by inducible NO synthase and cyclooxygenase-2, respectively. Protein tyrosine kinase-mediated phosphorylation and synthesis of any new proteins are likely to be required in this effect of VEGF in mouse skin. PMID- 9349635 TI - Comparison of the effects of nitric oxide synthase inhibition and guanylate cyclase inhibition on vascular contraction in vitro and in vivo in the rat. AB - We have investigated the differences between the nitric oxide synthase inhibitor (NOSI), L-NMMA, and the guanylate cyclase inhibitors (GCI), methylene blue and LY 83583, in their abilities to increase vasoconstrictor responses in vitro and in vivo. In rat small mesenteric arterial rings, 1 h exposure to the NOSI, L-NMMA (100 microM), and the GCI, methylene blue (10 microM), alone or in combination with L-NMMA, caused a significant reduction in the maximum relaxation to ACh in mesenteric arteries pre-contracted with the thromboxane mimetic U46619 (10 microM). Hence, both NOSI and GCI inhibit endothelium-dependent relaxations to ACh in rat small mesenteric artery. However, 1 h exposure to L-NMMA and L-NNA (both 100 microM), but not methylene blue (10 microM), significantly increased the contractile response to U-46619 (10 microM) in rat small mesenteric artery. It was decided to investigate further this difference between NOSI and methylene blue. In rat small mesenteric arterial rings, L-NMMA (10 microM) and LY 83583 (1 10 microM) significantly increased the contractile response to KCl (40 mM) or to noradrenaline (10 microM), when administered during the contraction. However, methylene blue (1-10 microM) increased the contractile response to KCl but not noradrenaline. In rat aortic rings, L-NMMA (100 microM), methylene blue (1-10 microM) and LY 83583 (1-10 microM) significantly increased the contractile response to KCl (40 mM) or to noradrenaline (1 microM). In the pithed rat preparation, L-NMMA (40.3 micromol kg(-1), i.v.) significantly increased the pressor response both to bolus injection of noradrenaline (3.13 nmol kg[-1]) and to spinal pressor nerve stimulation. However, methylene blue (3.13-15.6 micromol kg[-1]) or LY 83583 (4.0-40.0 micromol kg[-1]), failed to affect pressor responses to either NA or pressor nerve stimulation. Hence, there are differences between NOSI and GCI in their abilities to increase vasoconstrictor responses, especially when comparing responses in vitro and in vivo. This suggests that nitric oxide has actions in addition to activation of guanylate cyclase to modulate vasoconstrictor responses, presumably by membrane hyperpolarisation, and that this action may be more important in vivo. PMID- 9349637 TI - Biological activation of N(G)-nitro-D-arginine by kidney homogenate . AB - N(G)-nitro-L-arginine (L-NNA) and D-NNA have been shown to inhibit endothelium dependent relaxation. This study examined if the inhibitory effect of L-NNA or D NNA on relaxation is increased following incubation of the drug with the supernatant of tissue homogenates. Acetylcholine (ACh) caused concentration dependent relaxation of pre-constricted rat aortic rings with maximum relaxation of 95%. Maximum relaxations to ACh were reduced to 71 and 37% in the presence of D-NNA (40 microM) and L-NNA (1 microM), respectively. Relaxation to ACh was further reduced to 18% in the presence of D-NNA that was incubated for 1 h with the supernatant of kidney homogenate, but unaffected by D-NNA incubated with the supernatant of trichloroacetic acid-denatured kidney homogenate. Incubation of L NNA (1 microM) with either kidney supernatant or denatured kidney supernatant for 1 h did not affect its inhibitory effect on ACh-induced relaxation. Neither 1 h's incubation with plasma, or supernatants of liver, lungs or aorta homogenates affected the inhibitory action of D-NNA (40 or 120 microM) on ACh-induced relaxation. After D-NNA was incubated in kidney supernatant, its inhibitory effect on ACh-induced relaxation of the aorta was abolished by pretreatment of the aorta with L-arginine (L-Arg) but not D-Arg suggesting involvement of the L Arg pathway. The results suggest that D-NNA is converted by the kidney to a compound that acts similar to L-NNA. There appears to be little conversion of L NNA to D-NNA. PMID- 9349636 TI - Influence of different classes of NO synthase inhibitors in the pig gastric fundus. AB - The inhibitory potency of different classes of nitric oxide synthase (NOS) inhibitors (amino acid-based substances, guanidines, isothioureas, imidazoles and indazoles) versus peripheral neuronal NOS in the pig gastric fundus was investigated by studying their influence on electrically induced relaxations in non-adrenergic noncholinergic conditions. Circular muscle strips were mounted for isotonic registration in the presence of atropine and guanethidine, and tone was raised with 5-hydroxytryptamine. Electrical field stimulation (40 V, 0.1 ms, 4 Hz, 10 s) induced short-lasting relaxations. The inhibitory effect of 1 phenylimidazole could not be evaluated because it nearly abolished the 5 hydroxytryptamine-induced tone of the tissues. 7-Nitroindazole, imidazole, 2 iminobiotin and aminoguanidine did not inhibit the electrically induced relaxations, while the other 9 substances tested were able to do so. The influence of the incubation period was tested by studying the inhibitory effect after incubation for 10 up to 60 min. For N(G)-nitro-L-arginine methyl ester (L NAME), N(G)-nitro-L-arginine (L-NNA), L-N5-(1-iminoethyl)-ornithine (L-NIO), L-N6 (1-iminoethyl)-lysine (L-NIL), S-methyl-L-thiocitrulline and S-isopropyl isothiourea there was a moderate increase in the inhibitory effect up to 30 min of incubation so that they were incubated for 30 min to study their inhibitory potency. For L-thiocitrulline, S-methyl isothiourea and S-ethyl isothiourea, an incubation period of 60 min was used. The 9 substances concentration-dependently inhibited the electrically induced relaxations with a maximal inhibitory effect of approximately 80% except for S-methyl isothiourea (Emax of 53%). The overall order of potency was: S-isopropyl isothiourea > S-ethyl isothiourea > or = S methylL-thiocitrulline > or = L-NNA > L-NIO > L-NAME > S-methyl isothiourea > L thiocitrulline > L-NIL. While the potency for S-isopropyl isothiourea (EC50: 3.1 x 10(-5) M, n = 6) to S-methyl isothiourea (EC50: 11.5 x 10(-5) M, n = 5) was in the same range, the potency of L-thiocitrulline and L-NIL was clearly lower. This study showed several compounds to be potent inhibitors of peripheral neuronal NOS in the pig gastric fundus while some compounds, that were reported to inhibit brain neuronal NOS were not effective. The EC50 values found for the effective substrates in this functional study may be a guideline for the concentrations required to evaluate the role of NO in NANC neurotransmission in gastrointestinal smooth muscle preparations. PMID- 9349638 TI - Involvement of nitric oxide in the in vivo effects of lipopolysaccharide on the contractile and electrical properties of mouse diaphragm. AB - The contractile and electrical properties of the mouse diaphragm during endotoxemia were studied, and the possible role of nitric oxide (NO) on these changes was investigated. The mice were injected intraperitoneally with E. coli. lipopolysaccharide (endotoxin, LPS) at various times before isolation of the diaphragm to induce endotoxemia. It was observed that direct twitch tension was slightly increased, and that there was a significant increase in tetanic tension when compared with controls. The potentiation of direct twitch tension induced by a Cl--channel blocker (9-anthracene carboxylic acid), but not the potentiation by a Na+-channel activator (veratridine) or by K+-channel blockers (uranyl ion, 4 aminopyridine and tetraethylammonium ion), was attenuated in the diaphragm of LPS treated mice. Moreover, the resting membrane potential was significantly reduced and the membrane input resistance was significantly increased, largely due to a decrease in Cl--conductance. However, the membrane K+-conductance remained unaltered. These results imply that the sarcolemmal Cl--channel is markedly affected in the mouse diaphragm during endotoxemia. These changes of contractile and electrical characteristics of the mouse diaphragm during endotoxemia could be reversed by treatment with dexamethasone and N(G)-nitro-L-arginine (NO synthase inhibitors). On the other hand, in in vitro studies, LPS (20 microg/ml), by itself, applied directly to the diaphragm, did not alter the muscle contractions or the membrane potentials. A NO donor, added to the diaphragm bath, increased the tetanus/twitch ratio and induced a transient depolarization. All of these findings suggest that LPS may, at least in part, affect the sarcolemmal electrical properties and muscle contractions during endotoxemia through the L arginine:NO pathway. PMID- 9349640 TI - Gallium nitrate suppresses lupus in MRL/lpr mice. AB - Gallium (Ga) nitrate, a drug which prevents a variety of experimental autoimmune diseases, was investigated in a murine model of systemic lupus erythematosus (SLE). In one experiment, female MRL/Mp lpr/lpr (MRL/lpr) mice were randomized into 2 groups of 6: 1) vehicle (trisodium citrate) and 2) Ga. Subcutaneous injections began at 3 weeks of age and continued weekly until the mice were euthanized a week after the thirteenth injection. The loading dose of Ga (calculated as elemental Ga) was 45 mg/kg, followed by 15 mg/kg/week. In another experiment (n = 18) with 3 males and 3 females per group, mice received 1) vehicle, 2) Ga x 1 (one 45 mg/kg dose), and 3) Ga x 13. In the experiment with 12 mice, axillary lymph nodes from Ga-treated mice were significantly smaller than those from vehicle-treated mice (91+/-42 and 360+/-358 mg respectively, mean+/ SD), and spleens as well as lymph nodes from the former showed significantly less lymphoid infiltrate. In the experiment with 18 mice, prescapular lymph nodes weighed 312+/-98, 217+/-52, and 42+/-34 mg, and spleens weighed 732+/-492, 409+/ 164, and 192+/-93 mg in the groups which received vehicle, Ga x 1, and Ga x 13 respectively. Control mice had significantly more lymphoid infiltrates in the lungs, spleen, and lymph nodes and, unlike Ga x 13 mice, exhibited glomerulitis and renal vasculitis. Within groups, females developed more severe disease than males. The Ga x 13 group had increased percentages of CD4-bearing and CD8-bearing lymphocytes in lymph nodes and increased CD4-bearing lymphocytes in the spleen, with an increased proliferative response to mitogen stimulation in vitro in lymph nodes, although not in the spleen. The Ga x 13 group also gained less weight and developed osteosclerosis. Although preliminary, our findings suggest that clinical trials with Ga in SLE are merited. PMID- 9349641 TI - The ultradian EEG rhythm coincides temporally with the ultradian rhythm of histamine release in the posterior hypothalamus. AB - In the posterior hypothalamus of the rat, EEG power and release rates of several neurotransmitters oscillate according to ultradian rhythms. To investigate whether a causal relationship exists between EEG power and histamine release, the posterior hypothalamic area of the anaesthetized rat was superfused through a push-pull cannula combined with a tungsten electrode. Simultaneous EEG recording and determination of histamine release revealed that ultradian rhythms in the delta and theta frequency bands are negatively correlated to the release rate of histamine; periods of high neuronal activity, which might reflect synchronization of firing, coincide temporally with low release rate of histamine, while periods of low neuronal activity coincide with high histamine release rate. The alpha and beta frequency bands did not correlate with histamine release. The ultradian rhythm of EEG power and histamine release might be of importance for regulatory mechanisms, such as the secretion of hormones. PMID- 9349639 TI - Evidence for muscarinic M4 receptors mediating nonadrenergic noncholinergic relaxations in rabbit anococcygeus muscle. AB - The aim of the present study was to characterize the muscarinic receptor subtype mediating nonadrenergic noncholinergic (NANC) relaxations in the rabbit anococcygeus muscle (RAM) by the use of muscarinic receptor agonists and a battery of key muscarinic antagonists. In addition, experiments were carried out to identify the NANC neurotransmitter(s) involved in the inhibitory NANC neurotransmission. In preparations with histamine-raised tone, the nonselective muscarinic agonists (pD2 values) (+)-muscarine (5.23), cis-dioxolane (5.16), oxotremorine M (4.95) and carbachol (4.06) produced concentration-dependent relaxations corresponding to 72.6-85.0% of the histamine-induced precontraction. In contrast, the subtype-preferring (M1/M4 over M2 and M3 receptors) agonists 4 (3-chlorophenylcarbamoyloxy)-2-butynyltrimethylammonium chloride (McN-A-343), (S) 4-(dimethylamino)-1-methyl-2-butynyl-N-(3-chlorophenyl)carbamate methobromide [(S)-BN 228], (R)- and (S)-N-methyl-N-(1-methyl-4-pyrrolidino-2-butynyl)acetamide [(R)- and (S)-BM 5] showed no or rather low [(S)-BN 228] muscarinic activity. The low potencies, together with the ineffectiveness of some agonists, indicated a low effective receptor reserve associated with the relaxant response. No contractile responses to (+)-muscarine were observed neither in unstimulated nor in precontracted preparations suggesting that the existence of an excitatory postjunctional muscarinic receptor may be excluded. The nicotinic antagonist hexamethonium had no influence on relaxant responses to (+)-muscarine, but abolished relaxations elicited by (-)-nicotine. This demonstrates that the RAM contains also nicotinic acetylcholine receptors (AchRs) mediating inhibitory NANC responses. Relaxations induced by the stimulation of muscarinic and nicotinic AchRs as well as by electrical field stimulation (EFS) were completely abolished by tetrodotoxin and were also sensitive to the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine (L-NOARG), indicating that NO plays an important role as an inhibitory NANC transmitter in RAM. All muscarinic antagonists investigated did not influence the histamine-induced precontraction, but shifted the concentration-response curve of (+)-muscarine to the right in a parallel fashion. Schild analysis yielded regression lines of unit slope, indicating competitive antagonism. The following rank order of antagonist potencies (pA2 values) was found: 11-([4-[4-(diethylamino)-butyl]-1-piperidinyl]-acetyl-5,11 dihydro-6H-py rido (2,3-b) (1,4)-benzodiazepine-6-one hydrochloride (AQ-RA 741; 8.08) = himbacine (8.03) > or = tripitramine (7.69) > or = p-fluorohexahydro-sila difenidol (p-F-HHSiD; 7.48) > or = methoctramine (7.30) > or = pirenzepine (7.18) > or = guanylpirenzepine (6.24). A comparison of the pA2 values determined in the RAM with published data from binding studies at muscarinic M1-M4 and m5 receptors suggests that the functional muscarinic receptor mediating NANC relaxation in the RAM is of the M4 subtype. Taken together, the results obtained in the present study provide convincing evidence that relaxant responses elicited by muscarinic stimuli in RAM are nitrergic in nature and mediated by muscarinic M4 receptors located somadendritically on NANC neurons. Thus, the isolated RAM may serve as a functional muscarinic M4 receptor model. PMID- 9349642 TI - Stage duration and increase of work load in incremental testing on a cycle ergometer. AB - Any variation of the test protocol for incremental testing (IT) in cycle ergometry may affect the accuracy of the determination of anaerobic threshold (AnT). For lactate threshold concepts, indicating the maximum lactate steady state (max Lass), the formation of the quasi-steady-state (QSS=95% of steady state level) is evident. Previous studies have not specified the time that it takes for blood lactate to stabilize following incremental changes in WL. The purpose of this study was to identify the minimum duration of exercise necessary to establish QSS following various increments in WL (10, 20, 30, 40 and 50 W). Eight male endurance-trained cyclists [relative maximal oxygen consumption = 64.8 (4.2) ml x kg(-1) x min(-1)] performed three different exercise tests on a cycle ergometer: (1) an exhaustive IT with a starting WL of 100 W, followed by 20-W increments every 3 min; (2) a threshold test with 20-W increments every 9 min to determine the MaxLaSS; and (3) five incremental exercise tests (from 100/110 W, with 20-W increments every 3 min) with a final 10-, 20-, 30-, 40- or 50- W increment lasting 10 min, at 10 W below MaxLaSS (T10-T50 experiments). The time constant of lactate kinetics (tau), the time constant of lactate elimination, and the time taken to elicite QSS, defined as 95% of the time taken to reach steady state level (t95%), were calculated in the T10-T50 experiments. The tau and t95% increased significantly with WL increment size: the correlation was not linear. Smaller WL increments required proportionally longer durations. Mean (SD) t95% values (min:s) were 1:57 (0:27) (T10), 2:58 (0.16) (T20), 4:08 (0:23) (T30), 4:45 (0:45) (T40) and 5:06 (0:43) (T50). The application of these references in IT protocols may lead to an extension of total test duration, particularly with smaller increments. Therefore, lactate threshold modelling, the training status of the athletes and comparability with lactate measurements obtained during training events should be considered. IT protocols not accomplishing QSS criteria may effect an underestimation of WL-related lactate values and an overestimation of lactate thresholds, which indicate MaxLaSS, especially in highly trained athletes. This suggests that the establishment of an increment-size-dependent t95% may reduce protocol-related influences on AnT and standardize the use of the AnT in IT procedures in the training management of elite cyclists. PMID- 9349643 TI - Antioxidant status in various tissues of the mouse after fasting and swimming stress. AB - We studied the effect of fasting and swimming stress on a number of non-enzymatic and enzymatic antioxidant factors in various mouse tissues in order to see if their action was synergic. We examined levels of reduced (GSH), oxidized (GSSG) and total glutathione, total SH groups (TSH), sum of GSH and protein sulphydryl groups of cytosolic fractions, and the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase, glutathione reductase in adductor muscle, heart and liver. We also studied blood levels of GSH and glutathione bound to protein by mixed disulphides (GSSP). The case series consisted of four groups of animals (n = 10 for each group), namely no swimming and no fast, no swimming and fast, swimming and no fast, and swimming and fast. Fasting (18 h) resulted in a significant GSH depletion in all of the organs studied (-39% in the liver, -30% in the adductor muscle, -21% in the heart); GSSG increased significantly in the heart (+19%). Swimming to exhaustion, which lasted 3.95 (0.18) min [mean (SD), n = 10] with no significant difference between fast and no fast, resulted in a significant GSH depletion, to a percentage lower than that observed after fasting, in the adductor muscle and heart (-12% and -11%, respectively). In the blood of swimming mice, significant increases in GSH (+10%) and GSSG (+21%) levels were observed, whereas GSSP decreased (-15%). Enzyme activities after swimming were modified in only a few cases, and in a complex way. The findings of GSH depletion and a decrease in SOD activity in the adductor muscle seems to confirm the sensitivity of this organ to an overproduction of reactive oxygen species. At the same time, the GSSP decrease observed in blood was a new and unexpected finding, one that indicates a very prompt adaptation of red cells to increased oxidant states. PMID- 9349645 TI - Mechanomyography of the human quadriceps muscle during incremental cycle ergometry. AB - The mechanical activity of the human quadriceps muscle during maximal incremental cycle ergometry was investigated by mechanomyography (MMG). MMG and surface electromyography (EMG) recordings of vastus lateralis muscle activity were obtained from nine males. Cycle ergometry was performed at 60 rev/ min and work load was incremented step wise by 20 W (3.2 Nm) every minute until volitional fatigue. The mean amplitudes of MMG (mMMG) and EMG (mEMG) during the contraction phase were calculated from the last six contractions in each load. The duration, load and work rate of exercise at exhaustion were 13.3 (1.6) min, 44.1 (5.5) Nm, 276.7 (34.7) W, respectively. A linear relationship between mMMG and load was evident in each subject (r = 0.868-0.995), while mEMG seemed to dissociate as the load became greater. In the grouped mean data, mMMG was linearly related to load whether aligned to the absolute (r = 0.995) or maximal (r = 0.995) load. Involvement of the noise component was further investigated by studying passive cycling by four subjects. Pedals were rotated passively for the first half of each stage (PAS) and the subject then pushed the pedals for the second half (ACT). In the lighter load region, the mMMG of ACT was as small as that of PAS. However, the change in the mMMG of PAS was very small compared with that of ACT. In conclusion, this study demonstrates a linear relationship between the mMMG of the quadriceps muscle and work load during maximal incremental cycle ergometry. The effect of movement noise was thought to be small and stable. PMID- 9349644 TI - Fractional use of anaerobic capacity during a 30- and a 45-s Wingate test. AB - This study examined the suitability of the Wingate test as a means of assessing the maximal oxygen deficit (MOD), and the influence of the anaerobic capacity on the fraction of the MOD used during a 30- and a 45-s Wingate test in 19 male subjects. The MOD incurred in constant-intensity supramaximal exercise was higher (P < 0.01) than that for the 45-s and 30-s Wingate tests [68.6 (3.4) vs 60.9 (2.2) and 53.7 (1.6) ml x kg(-1), respectively], corresponding to a 10% higher value for the 45-s compared to that for the 30-s test (P < 0.001). A close correlation was found to occur between MOD and the oxygen deficit incurred during the 30- and 45-s Wingate tests, as well as between both all-out tests (r = 0.86 0.90; P < 0.001). The oxygen deficit accumulated during the first 30 s of the 45 s Wingate test was similar to that accumulated during the 30-s Wingate test. The intraclass correlation coefficient for the oxygen deficit after 30 s of all-out exercise (two treatments) was 0.96. The higher the MOD the lower was its fractional recruitment during the 30-s (r = -0.88, P < 0.001) and during the 45-s (r = -0.74, P < 0.01) Wingate tests. In conclusion, 80-90% as an assessment of the oxygen deficit incurred during a Wingate test is valid as an estimate of the anaerobic capacity. The fraction of the anaerobic capacity used in a 30- and 45-s all-out test is inversely related to the anaerobic capacity. PMID- 9349647 TI - Influence of long-latency reflex modulation on the performance of quick adjustment movements. AB - The effect of long-latency reflex modulation on the performance of a quick adjustment movement following a muscle stretch was studied in 26 healthy male subjects. When the subjects felt a sudden angle displacement in the direction of a wrist extension they were required to make an adjustment movement by moving a handlebar, held in the hand, to align with a target position as quickly and as accurately as possible. The index of performance (adjustment time) was the time taken to move the handle to the target position from stretch onset. A DC torque motor was used to evoke electromyographic (EMG) reflex responses on a wrist flexor. Averaging of the rectified EMG, recorded from surface electrodes placed over the flexor, showed short- and long-latency reflexes (M1 and M2 components). For all subjects, the amplitudes of the reflex components decreased during the adjustment movement because the target position for this study was fixed to the extension side of the wrist joint. The decrease in the M2 component, which is considered to be a transcortical reflex, was significantly larger than the decrease in the M1 component, which is spinal reflex. The main finding was of a positive correlation between the length of adjustment time and the degree of reduction of M1 and M2 with the adjustment movement (r = 0.602 for M1, P < 0.01; r = 0.850 for M2, P < 0.001). Moreover, there were correlations between the consistency of the voluntary response onset and the degree of M2 decrease (r = 0.577, P < 0.01), and between the consistency of the voluntary response onset and the length of the adjustment time (r = 0.603, P < 0.01). Therefore, we have concluded that the subjects who were able to perform adjustment movements within a short time could modulate the long-latency reflex of the muscle involved in such movements in order to make the function of their voluntary muscle activity more effective, and thus were able to respond appropriately. PMID- 9349646 TI - Effects of hydration state on hormonal and renal responses during moderate exercise in the heat. AB - The effects of hydromineral hormones and catecholamines on renal concentrating ability at different hydration states were examined in five male volunteers while they performed three trials. Each of these trials comprised a 60-min exercise bout on a treadmill (at 50% of maximal oxygen uptake) in a warm environment (dry bulb temperature, 35 degrees C; relative humidity, 20-30%). In one session, subjects were euhydrated before exercise (C). In the two other sessions, after thermal dehydration (loss of 3% body mass) which markedly reduced plasma volume (PV) and increased plasma osmolality (osm[pl]), the subjects exercised either not rehydrated (Dh) or rehydrated (Rh) by drinking 600 ml of mineral water before and 40 min after the onset of exercise. During exercise in the Dh compared to C state, plasma renin, aldosterone, arginine vasopressin (AVP), noradrenaline and adrenaline concentrations were increased (P < 0.05). A reduction in creatinine clearance and urine flow was also observed (P < 0.05) together with a decrease in urine osmolality, osmolar clearance and sodium excretion, while free water clearance increased (P < 0.05). However, compared to Dh, Rh partially restored PV and osm(pl) and induced a marked reduction in the time courses of both the plasma AVP and catecholamine responses (P < 0.05). Values for renal water and electrolyte excretion were intermediate between those of Dh and C. Plasma atrial natriuretic peptide presented similar changes whatever the hydration state. These results demonstrate that during moderate exercise in the heat, renal concentrating ability is paradoxically reduced by prior dehydration in spite of high plasma AVP levels, and might be the result of marked activation of the sympatho-adrenal system. Rehydration, by reducing this activation, could partially restore the renal concentrating ability despite the lowered plasma AVP. PMID- 9349648 TI - Functional subdivision of the upper trapezius muscle during low-level activation. AB - The electromyographic (EMG) amplitude was recorded using bipolar surface electrodes placed at different positions above the upper trapezius muscle of 16 healthy subjects. One of the aims of this study was to investigate the variation in EMG activity between electrode positions. For this purpose three tasks were performed: a mental activation test, a dynamic movement test and 90 degrees arm abduction. The EMG signals were full-wave rectified and averaged within windows that were 0.2 s in length. Normalized EMG activity showed significantly different EMG amplitudes at different electrode positions for two of the three tasks. The second aim of this study was to investigate whether the upper trapezius muscle may be functionally subdivided. For this purpose the normalized EMG amplitudes of each task were compared with the EMG amplitude recorded during submaximal shoulder elevation. While the EMG level was similar at one electrode position, significant differences were found at some of the other electrode positions, indicating a functional subdivision of the muscle. The present results indicate that for comparisons of upper trapezius EMG activity levels between some tasks or between subjects, it is worthwhile to make EMG recordings at several electrode positions. PMID- 9349649 TI - Mechanisms of underdeveloped sweating responses in prepubertal boys. AB - To approach the mechanisms underlying the underdeveloped sweating responses of prepubertal boys, 8 boys (7-11 years old) and 11 men (21-25 years old) were exposed to a standard heat stress for 60 min. The test consisted of placing the subjects' lower legs into a 42 degrees C water bath while they sat in otherwise constant environmental conditions (ambient temperature 25 degrees C and 45% relative humidity). Rectal (T[re]) and skin temperatures, local sweating rates (mSW: on the chest, back, forearm and thigh) and the frequency of sweating expulsions (fSW: as an indicator of central sudomotor activity) were measured during the test. During the passive heating, no group differences were observed for the increase in T(re), mean skin temperature and metabolic heat production. However, mean body temperature (Tb) during heating was significantly higher for the boys (P < 0.001) because of a higher baseline T(re). The boys had lower mSW on the chest (P < 0.004) and thigh (P < 0.001) during the latter half of the 60 min exposure compared to the young men, although a similar mSW was observed between the groups during the first half of the test. The group difference of mSW on the back was similar to that of the chest and thigh, but the difference was not significant (P = 0.10). In contrast, the boys had a greater mSW on the forearm throughout the heating (P < 0.03). The slope of the mSW vs fSW relationship was significantly lower for the chest and thigh in the boys compared to the men (P < 0.05), and the same tendency was observed for the back (but was not significant, P = 0.10). In contrast, no difference was observed between the groups for the slope of mSW vs fSW for the forearm. Furthermore, a lower sweat output per gland was also observed on the chest, back, and thigh in the boys (P < 0.01), but not on the forearm. No group difference was observed for the slope of the fSW vs Tb relationship. These results suggest that the lower mSW observed in the prepubertal boys were due possibly to underdeveloped peripheral mechanisms, including the sweat glands and their surrounding tissues, rather than to an underdeveloped central drive activity related to sudomotor function. Regional differences may well exist in any underdeveloped peripheral mechanism associated with maturation. PMID- 9349650 TI - Adequacy of food rations in soldiers during exercise in hot, day-time conditions assessed by doubly labelled water and energy balance methods. AB - The energy requirements of people doing physical work in hot climates are not clearly understood. In particular, we know little about the combined effects of heat stress and muscular work on energy requirements. During military exercises in the African bush, soldiers are supplied with standard rations, the adequacy of which is unknown. We have now assessed the adequacy of these food and water rations in 12 male Zimbabwean soldiers during 12 days of strenuous, heat-stress exercise in the field. We used two methods to measure energy expenditure: the double-labelled water method (DLW) and the energy balance method (i.e. comparing dietary energy with changes, if any, in body energy stores). Two groups were studied: one group (eight subjects) carried out field exercises; the control group consisted of four soldiers doing normal work. Mean daily energy expenditure as assessed by the DLW method was [mean (SE)] 23 (1.5) MJ x day(-1) for the field group and 14 (0.5) MJ x day(-1) for the control group (P < 0.001). By the energy balance method, daily energy expenditure was calculated to be 26 (0.7) MJ x day( 1) and 15.5 (0.4) MJ x day(-1) for the field group and control group, respectively. Body mass loss was 3 (0.1) kg [4.6 (0.3)% of body mass] for the field group, but the control group gained 1.1 (0.1) kg. Mean daily fluid intake was 11 (0.5) 1 x day(-1), suggesting that the standard ration supplied was inadequate. Body mass loss was caused by both the energy deficit and total body water loss. These results suggest strenuous work in hot, dry field conditions imposes extra energy requirements. PMID- 9349651 TI - Expired air temperature during prolonged exercise in cool- and hot-humid environments. AB - The purpose of this investigation was to measure expired air temperature under cool- and hot-humid environmental conditions at rest and during prolonged exercise to: (1) establish if significant increases in body core temperature affected expired air temperature, and (2) to determine if the temperature setting for heating the pneumotachometer in an open-circuit system requires adjustment during prolonged exercise tests to account for changes in expired air temperature. Six male distance runners completed two tests in cool-humid [dry bulb temperature (Tdb) 15.5 (SD 1.3) degrees C, wet bulb temperature (TWb) 12.1 (SD 1.4) degrees C] and hot-humid [Tdb 31.6 (SD 0.6) degrees C, TWb 24.9 (SD 0.6) degrees C, black globe temperature (Tg) 34.3 (SD 0.3) degrees C] environments, running at a velocity corresponding to 65% [67.1 (SD 2.82)%] of their maximal oxygen uptake. Rectal temperature and expired air temperatures were compared at rest, and after 30 min and 60 min of exercise for each environment. The main finding of this investigation was a significant (P < 0.05) but small increase in expired air temperature between the 30-min and 60-min measures in the hot-humid environment. No significant differences in expired air temperature were found between the 30-min and 60-min measures in the cool-humid environment. These findings suggest that: (1) expired air temperature is influenced by elevations in body core temperature during prolonged exercise in hot-humid conditions, and (2) that the temperature setting for heating the head of the pneumotachometer (after determining the appropriate temperature through measuring expired air temperature for the set environmental condition) may require adjustment during prolonged exercise trials in hot-humid environmental conditions. PMID- 9349652 TI - Effects of acute expansion of plasma volume on cardiovascular and thermal function during prolonged exercise. AB - To investigate the hypothesis that an increase in plasma volume (PV) is obligatory in reducing the cardiovascular drift that is associated with prolonged exercise following training, a plasma expander (Macrodex) was used to acutely elevate PV. Eight untrained volunteers [maximal oxygen consumption; VO2max 45.2 (2.2) ml x kg(-1) x min(-1), mean (SE)] cycled for 2 h [at 46 (4)% VO2max] in ambient conditions either with no PV expansion (CON) or following PV expansions of either 14% (LOW) or 21% (HIGH). During CON, heart rate (HR) increased (P < 0.05) from 147 (2.4) beats x min(-1) to 173 (3.6) beats x min(-1) from 15 to 120 min of exercise. Both LOW and HIGH conditions depressed (P < 0.05) HR, an effect that was manifested following 15 min of exercise. In contrast, stroke volume (SV) was elevated following PV expansion, with values (ml) of 89.6 (6.8), 97.8 (5.9) and 104 (4.6) noted by 15 min of exercise for CON, LOW and HIGH conditions, respectively. Acute PV expansion, regardless of magnitude, also resulted in elevations in cardiac output (Qc). These differences between conditions persisted throughout the exercise, as did the elevation in Qc that was noted with LOW and HIGH conditions. No difference between Qc, HR or SV was found between LOW and HIGH. In addition, neither LOW nor HIGH conditions altered the change in rectal temperature that was observed during exercise. These results demonstrate that, at least for moderate exercise performed in ambient conditions, PV expansion serves only to alter cardiac function (Qc, HR, SV) early in exercise, and not to attenuate the drift that occurs as the exercise is prolonged. PMID- 9349653 TI - Mechanomyography and oxygen consumption during incremental cycle ergometry. AB - The purpose of this investigation was to describe and compare the relationships for mechanomyography (MMG) and oxygen consumption rate (VO2) versus power output during incremental cycle ergometry. Twenty four adult males [mean (SD) age, 22.1 (2.0) years] volunteered to perform an incremental test to exhaustion on a cycle ergometer. A MMG piezoelectric recording device was placed mid-thigh over the vastus lateralis muscle and VO2 was measured using standard open circuit procedures. The r2 values for the MMG and VO2 versus power output relationships ranged from 0.79 to 0.99 and 0.97 to 0.99, respectively. In 20 of the 24 subjects there was no significant (P > 0.10) difference between the slope values for the normalized MMG and VO2 (expressed as a percentage of maximal values) versus power output relationships. The results of this study indicate that MMG procedures can be used to quantify muscular activity and monitor changes in exercise intensity during cycle ergometry. Furthermore, the present findings demonstrated a close association between the mechanical (MMG) and metabolic (VO2) aspects of muscular contraction during incremental cycle ergometry. PMID- 9349654 TI - Neuromuscular control following maximal eccentric exercise. AB - Kinematic and electromyographic (EMG) analysis of a target-directed, maximal velocity movement was used to investigate the effects of high-force eccentric exercise on the neuromuscular control of elbow flexion. Ten non-weight-trained females [19.6 (1.6) years old] performed 50 maximal velocity elbow flexion movements from 0 to 1.58 rad (90 degrees), as rapidly as possible in response to a light stimulus, while kinematic and triphasic EMG parameters were measured. This was done three times pre-exercise, immediately and 1, 2, 3, 4, and 5 days following the 50 maximal eccentric elbow flexion actions. The eccentric exercise caused lengthening of kinematic parameters including total movement time and time to peak velocity. The EMG elements of the biceps brachii (b.) motor time, time to peak EMG, biceps b. burst duration, and the latency period between biceps b. and triceps b. bursts were lengthened post-exercise. These changes persisted for up to 5 days post-exercise. The exercise also caused a large increase in serum creatine kinase (CK) activity. It was concluded that high-force eccentric exercise in this population caused prolonged changes in neuromuscular control that were a function of exercise-induced disruption of the skeletal muscle. Compensation in the central motor program was such that the components of the triphasic EMG pattern were systematically lengthened. PMID- 9349655 TI - Torque-velocity relationship during cycle ergometer sprints with and without toe clips. AB - The torque-velocity relationship in cycling has been studied during all-out sprints (n = 6 subjects) with and without toe clips on an electronic Lode ergometer with strain gauges, to estimate the importance of the expected decrease in torque, velocity and power output. As previously found with different cycling protocols, the torque-velocity relationship was linear for all-out sprints with toe clips. A similar relationship was observed when cycling without toe clips but the torque-velocity relationship was inflected downwards at low or high velocities in several subjects who were not regular cyclists. The pulling action during the rise of the pedal at low velocities cannot explain why the torque velocity relationship is not hyperbolic for cycling exercises with toe clips because similar relationships were observed without toe clips. The maximal power output was significantly higher during cycling with toe clips (782 W vs 668 W, P < 0.05), probably because of the pulling action at low and medium velocities as indicated by the higher value of the extrapolated maximal torque T0 (138 N x m vs 122 N x m, P < 0.05). In contrast, the maximal extrapolated velocity, V0 and peak velocity were not significantly improved by the use of toe clips. The comparison of the angle-torque patterns at low and high velocities suggested that the kinetic energy of the legs can be transformed into power output when cycling without toe clips as well as it can when cycling with toe clips. PMID- 9349656 TI - Effects of artificially-induced anaemia on sudomotor and cutaneous blood flow responses to heat stress. AB - The influence of artificially induced anaemia on thermal strain was evaluated in trained males. Heat stress trials (38.6 degrees C, water vapour pressure 2.74 kPa) performed at the same absolute work rates [20 min of seated rest, 20 min of cycling at 30% peak aerobic power (VO2pcak), and 20 min cycling at 45% VO2peak] were completed before (HST1) and 3-5 days after 3 units of whole blood were withdrawn (HST2). Mild anaemia did not elevate thermal strain between trials, with auditory canal temperatures terminating at 38.5 degrees C [(0.16), HST1] and 38.6 degrees C [(0.13), HST2; P > 0.05]. Given that blood withdrawal reduced aerobic power by 16%, this observation deviates from the close association often observed between core temperature and relative exercise intensity. During HST2, the absolute and integrated forearm sweat rate (mSW) exceeded control levels during exercise (P < 0.05), while a suppression of forehead mSW occurred (P < 0.05). These observations are consistent with a possible peripheral redistribution of sweat secretion. It was concluded that this level of artificially induced anaemia did not impact upon heat strain during a 60-min heat stress test. PMID- 9349657 TI - Inhibition of M-type K+ and N-type Ca2+ channels by the human gonadotropin releasing-hormone receptor heterologously expressed in adult neurons. AB - Gonadotropin-releasing hormone (GnRH) controls all aspects of reproductive function. GnRH is secreted by hypothalamic neurons and exerts its effects on the endocrine system through pituitary gonadotropes, while its effects on sexual receptivity are mediated by the central nervous system. The electrophysiological responses of central neurons to GnRH have shown both excitatory and inhibitory responses, but little is known about the mechanisms by which GnRH can change neuronal excitability. The present study addresses the mechanisms whereby stimulation of the human GnRH receptor changes neuronal excitability by using a combination of electrophysiological and heterologous expression techniques. Microinjection of in vitro transcribed cRNA coding for the human GnRH receptor into enzymatically dissociated adult rat superior cervical ganglion neurons resulted in GnRH receptor expression. Activation of the GnRH receptor inhibited both M-type K+ and N-type Ca2+ channels. Inhibition of M-type K+ channels was insensitive to pertussis toxin pretreatment and blocked by intracellular GDPbetaS. Inhibition of Ca2+ channels was slow in onset, voltage independent and insensitive to pertussis toxin. Wash-out of GnRH resulted in an unusual transient reversal of tonic G-protein-mediated Ca2+ channel inhibition. Block of the N-type Ca2+ channel with omega-conotoxin GVIA decreased Ca2+ current inhibition from 43 to 15%, indicating that the N-type Ca2+ channel is an effector target. Ca2+ channel inhibition was completely abolished by including a Ca2+ chelator in the patch pipette. Cell-attached macropatch experiments indicated that Ca2+ channel inhibition is mediated by a diffusible second messenger. These results demonstrate that the human GnRH receptor can inhibit M-type K+ and N-type Ca2+ channels when heterologously expressed in adult rat neurons. Modulation of M-type K+ and N-type Ca2+ channels in central neurons which contain GnRH receptors is likely to contribute to the changes in neuronal excitability elicited by GnRH. PMID- 9349658 TI - Regulation of gonadotropin-releasing hormone and luteinizing hormone secretion by AMPA receptors. Evidence for a physiological role of AMPA receptors in the steroid-induced luteinizing hormone surge. AB - Recent work has demonstrated that glutamate functions as a major transmitter involved in the regulation of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion in female animals, although the specific receptors and mechanisms mediating its effects have not been completely worked out. The purpose of the present study, therefore, was to examine the role of the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid)-type glutamate receptor in the control of GnRH and LH secretion in female animals. Toward this end, the dose- and steroid-dependent effects of AMPA on GnRH and LH secretion in female rats were investigated using both in vitro and in vivo approaches, and the role of AMPA receptors in the production of the steroid-induced LH surge was also assessed. The results of the study revealed that central administration of AMPA resulted in a stimulation of LH release in the estrogen-primed ovariectomized adult rat. AMPA was also found to potently stimulate GnRH release in vitro from mediobasal hypothalamic (MBH) fragments obtained from estrogen-primed ovariectomized adult rats, and this effect was blocked by the selective AMPA receptor antagonist, NBQX. The mechanism of action of AMPA appeared to differ from that of N-methyl-D-aspartate (NMDA) as AMPA, in contrast to NMDA, failed to elevate nitric oxide synthase activity in the hypothalamus. The effect of AMPA on LH secretion was demonstrated to be steroid dependent, as central administration of AMPA stimulated LH release in estrogen-primed ovariectomized rats, but was inhibitory to LH release in non-estrogen-primed ovariectomized rats. In contrast, AMPA stimulated GnRH release equally well from MBH fragments obtained from estrogen-primed or non-estrogen-primed ovariectomized rats. The different effects of AMPA on LH release may be due to different pituitary sensitivities between the two models, or alternatively, AMPA may stimulate the release of LH inhibitory factors in the ovariectomized rat in the absence of estrogen. Finally, a physiological role for AMPA receptors in the production of the steroid-induced LH surge was suggested, based on the finding that central administration of the selective AMPA receptor antagonist, NBQX, into the third cerebroventricle significantly attenuated the steroid-induced LH surge in the ovariectomized adult female rat. PMID- 9349659 TI - Stimulatory effect of melanin-concentrating hormone on luteinising hormone release. AB - Alpha-melanocyte-stimulating hormone (alpha-MSH) and melanin-concentrating hormone (MCH) are two peptide neurotransmitters widely distributed in the mammalian brain, the former originating mainly from cell bodies in the arcuate nucleus and the latter from cell bodies in the zona incerta and lateral hypothalamus. Within the hypothalamus they innervate the pre-optic area, median eminence (ME) and ventromedial nucleus (VMN). Both peptides stimulate sexual behaviour and in this report we have investigated their effect on another gonadal steroid-dependent function, luteinising hormone (LH) release. alpha-MSH, MCH or a combination of the two were injected bilaterally (100 ng/side) into either the medial pre-optic area (MPOA), ME, or VMN of anaesthetised (Saffan 3 ml/kg i.p.) rats that had previously been ovariectomised and adrenalectomised (O+A) and then primed with 5 microg/rat s.c. oestradiol benzoate (OB), 48 h before peptide administration. MCH stimulated LH release when applied to the MPOA and ME; alpha MSH was inhibitory in the ME and in this model was ineffective in the MPOA. Neither peptide was effective in the VMN. The two peptides were then injected into the MPOA of O+A rats primed with OB followed 48 h later by 0.5 mg/rat s.c. progesterone, which normally induces an LH surge. alpha-MSH, but not MCH, inhibited this induced rise in LH. Administration of anti-MCH antiserum (0.5 microg/side neat serum) also had an inhibitory effect on LH release in this model. These results show that MCH has a stimulatory effect on LH release when administered into the ME and MPOA. In the MPOA, this may be physiologically significant since blocking endogenous MCH with an anti-MCH antiserum inhibits LH release. On the other hand, alpha-MSH has an inhibitory effect on LH release in the MPOA and ME. In the teleost skin these two peptides are functionally antagonistic; it seems that a similar antagonism exists between their effects on LH release. PMID- 9349660 TI - Luteinising hormone secretion from the perifused ovine pars tuberalis and pars distalis: effects of gonadotropin-releasing hormone and melatonin. AB - It is not known where melatonin acts to influence the neuroendocrine axis of seasonally breeding mammals. However, since the pars tuberalis (PT) contains the highest density of melatonin receptors, this adenohypophyseal subdivision is a potential target. Gonadotropes are the only immunocytochemically detectable adenohypophyseal cell type of abundance in the PT. This study investigated whether melatonin could modulate basal and/or gonadotropin-releasing hormone (GnRH)-stimulated luteinising hormone (LH) secretion from the ovine PT and pars distalis (PD) in vitro. Tissue fragments from both pituitary areas were placed in separate chambers in a constant-environment perifusion system (37 degrees C; 100 microl/min) and 10-min effluent fractions were collected, frozen and later assayed for LH. Sixty minutes prior to a GnRH challenge (10 min; 10 nM), melatonin (1 microM or 100 nM) was added to the perifusate of half the tissue fragments. GnRH increased (p < 0.01) LH output from both pituitary subdivisions. Melatonin attenuated (p < 0.05) the GnRH-induced increase in LH output from the PT but not from the PD. The physiological importance of this melatonin-attenuated PT LH is unknown but it may play a role in modulating the neuroendocrine reproductive axis. PMID- 9349661 TI - Modulation by galanin of growth hormone and gonadotropin secretion from perifused pituitary and median eminence of prepubertal male calves. AB - Galanin is widely distributed in the peripheral and central nervous system and has been indicated as a putative hypothalamic-hypophysiotropic hormone. This study was performed to investigate the effects of galanin on both growth (GH) and luteinizing hormones (LH) from pituitaries of young male calves. Pituitary slices (P, 500 microm in thickness) were perfused alone or coincubated with median eminence terminals (ME) in DMEM-F12 plus BSA 0.1% and antibiotics. The perifusion chambers were kept in equilibrium for 150 min, and medium samples were collected every 10 min for 240 min and stored at -20 degrees C until the measurement of LH and GH levels. Basal GH release increased up to 60% after galanin infusion (p < 0.01 vs. baseline levels) for 60 min in P alone; in P + ME coincubation, galanin stimulated GH secretion was further increased by up to 200%. Basal LH release in chambers with P was significantly increased (up to 25%; p < 0.05) for 70 min after galanin infusion; P + ME coincubation showed a galanin-mediated increase in LH release of up to 50%. GH and LH responsiveness to exogenous GH-releasing hormone and gonadotropin-releasing hormone was not significantly modulated by galanin in our experimental model. In conclusion, galanin is demonstrated to have a significant stimulatory role in the secretion of GH and LH, with a combined action at both the hypothalamic and pituitary levels. ...................... PMID- 9349662 TI - Prolonged oral treatment with MK-677, a novel growth hormone secretagogue, improves sleep quality in man. AB - Previous studies have indicated the existence of common mechanisms regulating sleep and somatotropic activity. In the present study, we investigated the effects of prolonged treatment with a novel, orally active, growth hormone secretagogue (MK-677) on sleep quality in healthy young and older adults. Eight young subjects (18-30 years) followed a double-blind, placebo-controlled, three period crossover design. Each subject participated in three 7-day treatment periods (with bedtime drug administration), presented in random (Latin square) order, and separated by at least 14 days. Doses were 5 and 25 mg MK-677 and matching placebo. Six older subjects, ages 65-71 years, each participated in two 14-day treatment periods (with bedtime drug administration) separated by a 14-day washout. Doses were 2 and 25 mg MK-677 during the first and second periods, respectively. Baseline sleep and hormonal data were obtained on the 2 days preceding the beginning of the first 14-day treatment period. In young subjects, high-dose MK-677 treatment resulted in an approximately 50% increase in the duration of stage IV and in a more than 20% increase in REM sleep as compared to placebo (p < 0.05). The frequency of deviations from normal sleep decreased from 42% under placebo to 8% under high-dose MK-677 (p < 0.03). In older adults, treatment with MK-677 was associated with a nearly 50% increase in REM sleep (p < 0.05) and a decrease in REM latency (p < 0.02). The frequency of deviations from normal sleep also decreased (p < 0.02). The present findings suggest that MK-677 may simultaneously improve sleep quality and correct the relative hyposomatotropism of senescence. PMID- 9349663 TI - Increased vasopressinergic activity as a possible compensatory mechanism for a normal hypothalamic-pituitary-adrenal axis response to stress in BALB/c nude mice. AB - A bidirectional relationship between the immune and neuroendocrine systems is now widely accepted. Since it is well known that the thymus plays an important role in the regulation of the immune function, we decided to explore whether a lack of the thymic function may influence hypothalamic-pituitary-adrenal (HPA) axis activity. Eight-week-old female mice of both strains, nude and control BALB/c, were used to study: (a) the in vivo response of the HPA axis to several stress stimuli acting at either the hypothalamic (insulin administration and ether vapor inhalation), pituitary (CRH and vasopressin injections) or adrenal (ACTH treatment) level and (b) the in vitro response of hypothalamic fragments to high KCl (48 mM) stimulation. The results indicate that: (1) basal plasma ACTH and vasopressin levels were significantly (p < 0.05) higher in nude than in control BALB/c mice, whereas basal plasma corticosterone concentrations were similar in both strains of mice; (2) although no significant strain-related difference in the stress-induced ACTH secretion in plasma was found, hypothalamic stimuli were able to induce a significantly (p < 0.05) higher secretion of glucocorticoid in plasma in nude than in control BALB/c mice; (3) the pattern of in vitro hypothalamic CRH release was similar in both strains of animals; however, basal AVP output and that stimulated by 48 mM KCl were significantly (p < 0.05) higher in nude than in control hypothalamic fragments, and (4) whereas hypothalamic CRH, pituitary ACTH and adrenal glucocorticoid contents were similar in both strains, hypothalamic AVP content was significantly (p < 0.05) higher in athymic than in control mice. In summary, our results indicate that nude mice have an increased vasopressinergic function which could contribute to a normal HPA axis activity; thus, adult athymic mice of BALB/c origin could compensate, due to their increased vasopressinergic function, for a robust glucocorticoid release to protect themselves immediately after aggression. It remains to be determined whether this enhanced vasopressinergic function is a result of an early adrenal insufficiency due to congenital deficiency of thymic factors known to stimulate HPA axis function. PMID- 9349664 TI - Distribution and expression of glucocorticoid receptor mRNA in the forebrain of the rainbow trout. AB - The expression and distribution of glucocorticoid receptor mRNA was studied in the forebrain of mature female and immature undifferentiated rainbow trout (Oncorhynchus mykiss) by means of Northern blotting and in situ hybridization. A single mRNA species of 7.5 kb was detected in mRNA polyA+ prepared from the anterior brain. In situ hybridization was carried out using a 35S-labelled riboprobe corresponding to the A/B-domain (between nucleotides 1224 and 1763) of the recently cloned rainbow trout glucocorticoid receptor cDNA. Comparison of adjacent sections hybridized with the sense and antisense probes allowed detection of a specific signal with a similar distribution pattern in all animals studied. In the telencephalon, a specific hybridization was detected in scattered cells of the dorsal telencephalic hemisphere, but the stronger signal was consistently observed in the dorsal nucleus, and to a lesser degree in the ventral nucleus of the ventral telencephalon. Heavy hybridization staining was consistently observed in all subdivisions of the preoptic nucleus and the nucleus lateralis tuberis, which are the main hypophysiotrophic regions in fish. A weaker signal was detected in the nucleus anterioris periventricularis, nucleus suprachiasmaticus and thalamic region. The presence of a strong signal in virtually all magnocellular neurons of the nucleus preopticus, known for producing vasotocin, isotocin and corticotropin-releasing factor favors a direct function of cortisol in regulating its own secretion. PMID- 9349665 TI - Induction of the expression of gag protein in HTLV-I infected lymphocytes by anti ICAM 1 antibody in vitro. AB - Intercellular adhesion molecule 1 (ICAM 1) is an inducible protein ligand which is up-regulated during inflammation and is either not constitutively expressed or is only expressed at low levels. The expression of ICAM 1 increases in HTLV-I infected T cell lines as well as in CD4+ T-cells of peripheral blood lymphocytes (PBLs) from HAM/TSP patients. After PBLs of HAM/TSP patients were cultured in the presence of stimulating anti-ICAM 1 antibody, the expression of the HTLV-I gag protein in PBLs was observed by both immuno-histostaining and western blot analysis using HTLV-I Ag-specific mouse monoclonal antibody. These data thus suggested that the signal transduction via adhesion molecule, ICAM 1 could induce the transcription of the HTLV-I gene and this might therefore play an important role in the pathogenesis of HAM/TSP. PMID- 9349666 TI - Expression of cell adhesion molecules in normal nerves, chronic axonal neuropathies and Schwann cell tumors. AB - Cell adhesion molecules (CAMs) play a role in the normal development and regeneration of tissues as well as in the biological behaviour of tumors. We studied the immunohistochemical expression of various CAMs, such as neural cell adhesion molecule (NCAM), its polysialylated isoform (PSA-NCAM), epithelial (E-) cadherin, and beta1 integrins (alpha2beta1, alpha5beta1, alpha6beta1) in a series of frozen specimens of 10 normal nerves, 5 axonal neuropathies, 26 benign Schwannomas and 2 malignant peripheral nerve sheath tumors (MNST). NCAM was expressed by non-myelinating Schwann cells from normal nerves and overexpressed by Schwann cells from patients with chronic axonal neuropathies and Schwannomas. The expression was lower in MNST. Expression of PSA-NCAM was heterogeneously displayed by Schwann cells from the various tissues studied. Anti E-cadherin immunoreactivity was present in myelin sheath in normal nerves and axonopathies. It was expressed in some Schwannomas especially in vestibular Schwannomas. Integrins VLA alpha2 and VLA alpha6 were widely expressed by Schwann cells from normal nerves, axonal neuropathies and Schwannomas but their expression was low in MNST. VLA alpha5 was not expressed by Schwann cells from normal nerve and Schwannomas but present in chronic axonal neuropathies and MNST. In addition VLA alpha6 was strongly expressed by perineurial cells. These data show that CAMs have a characteristic pattern of expression in normal nerve. Also, some CAMs are always expressed by Schwann cells but the expression of others differs in normal nerves versus axonopathies or tumors, suggesting a role of the microcellular environment in the regulation of CAM expression. Schwannomas have different pattern of expression than MNST. PMID- 9349667 TI - Mitochondrial mobility in differentiating muscle heterokaryons. AB - Ragged-red fibers, a morphological hallmark of many patients with mitochondrial encephalomyopathies who harbor mitochondrial DNA (mtDNA) mutations, usually contain varying ratios of mutated and wild-type mtDNAs. Deficient respiratory function in muscle is almost invariably segmental. To investigate whether this observation may be explained by restricted lateral movement of mitochondria within myofibers, we studied the spatial and temporal behavior of two different mitochondrial populations within multinucleate myotubes. We co-cultured normal human and mouse myoblasts, allowed them to fuse into muscle heterokaryons and investigated whether the mitochondria remained segregated, or migrated and intermixed. Human and mouse nuclei were identified by their differential staining pattern with the dye Hoechst 33 258 and mitochondria were distinguished immunologically and by in situ hybridization. Although we observed some territoriality at very early time points after myoblast fusion, there was rapid intermixing of the mitochondrial populations, as early as 48 h after myoblast fusion. We conclude that mitochondria, unlike many other muscle components, lack territorial organization in cultured, differentiating heterokaryons. PMID- 9349668 TI - Myosin-induced autoimmune polymyositis in the rat. AB - Experimental autoimmune myositis (EAM) was induced in Lewis rats by immunization with partially purified and purified skeletal myosin. Although clinical signs such as muscle weakness were very mild, multiple inflammatory lesions in the skeletal muscle, but not in the heart, were found by histological examination. Immunohistochemical staining revealed that muscle fiber-infiltrating cells were CD8+ and CD11b+ cells and that CD4+, TCR alphabeta+, B and NK cells were mainly located in the endomysium and interfiber connective tissue. These findings were in contrast to those obtained in experimental autoimmune encephalomyelitis lesions in which CD4+ cells predominate over CD8+ cells. T cells and sera isolated from myosin-immunized animals responded vigorously to myosin. However, neither sensitized lymphoid cells mainly comprising CD4+ cells nor purified anti myosin immunoglobulin G mediated the disease into naive rats, suggesting that T cells other than CD4+ cells such as CD8+ cells may be the final effector. Taken together, EAM induced in Lewis rats is similar to human polymyositis (PM). EAM can serve as a good model for human PM and give insight into the pathogenesis of the disease. PMID- 9349669 TI - Positive and negative factors in movement control: a current review of Denny Brown's hypothesis. AB - In his extensive writings, Denny-Brown hypothesized that two competitive 'tropisms,' one positive (exploratory) and one negative (withdrawal) act to coordinate normal movements at all levels of the neuraxis. Lesions in particular areas of the central nervous system result in disequilibrium between these tropisms, leading to disorders of posture and movement, including involuntary movements. The tactile manifestations of unbalanced exploratory tropisms are grasping responses, whereas the complementary withdrawal tropisms are avoiding responses. In Denny-Brown's view, at the level of the cerebral cortex, grasping responses result from frontal lobe injury whereas avoiding responses result from parietal lobe lesions. In this report we review Denny-Brown's conceptions of positive and negative tropisms, their anatomical loci, and whether his hypothesis has merit in a contemporary approach to brain function. We find that Denny Brown's view on the anatomical loci associated with these behaviors is incomplete, but that the idea of conflicting behavioral tendencies is valuable for understanding and managing some neurological and perhaps also psychiatric disorders. For example, his hypothesis offers an important perspective in understanding the paradoxical success of stereotaxic surgery to alleviate the symptoms of Parkinson's disease. PMID- 9349670 TI - Changes in the concentration of amino acids in serum and cerebrospinal fluid of patients with Parkinson's disease. AB - The concentrations of sixteen amino acids have been measured in the serum and cerebrospinal fluid (CSF) of patients with Parkinson's disease and compared with those of control subjects. The levels of most amino acids were not different between the two groups, but the level of glutamate in CSF was decreased significantly, while the level of glutamine was increased. The results may be consistent with an alteration of glutamate neurotransmission in Parkinson's disease. PMID- 9349671 TI - Alteration of cytokine levels by fosfomycin and prednisolone in spontaneous proliferation of cultured lymphocytes from patients with HTLV-I-associated myelopathy (HAM/TSP). AB - Fosfomycin has recently been reported as an antibiotic with immunomodulatory activities. To evaluate the possibility of clinical administration of fosfomycin in patients with human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP), the effects of this agent on the HTLV-I-induced in vitro phenomenon were studied. The influence of fosfomycin on in vitro spontaneous proliferation (SP) of peripheral blood mononuclear cells (PBMCs) from four patients with HAM/TSP was measured by thymidine incorporation into the cells, and the concentration of several cytokines in the culture supernatants was examined in three HAM/TSP patients. Enzyme-linked immunosorbent assays (ELISAs) were employed to detect the concentrations of interleukin-4 (IL 4), IL-6, IL-10, interferon-gamma (IFN-gamma), transforming growth factor-beta1 (TGF-beta1), and macrophage inflammatory protein-1alpha (MIP-1alpha). The data were compared to the changes by prednisolone which is known to regulate the HTLV I-associated in vitro phenomenon and to have a therapeutic benefit in patients with HAM/TSP. Production of IL-6, IFN-gamma and MIP-1alpha from the spontaneously proliferating cells were demonstrated. Fosfomycin could not suppress the HTLV-I associated SP, but had the properties to decrease the levels of TGF-beta1 and MIP 1alpha. It was also demonstrated that the concentrations of IFN-gamma and MIP 1alpha in the cultures in the presence of prednisolone were apparently decreased, suggesting a possible involvement of these cytokines in the pathogenesis of HAM/TSP. These findings support the hypothesis that fosfomycin may have immunomodulatory potentials in HTLV-I-related cellular interactions in a different manner from ordinary immunomodulatory agents. PMID- 9349672 TI - Analysis of single-joint rapid movements in patients with sporadic olivopontocerebellar atrophy. AB - Patients with pure cerebellar cortical atrophy (CCA) present isolated cerebellar signs, whereas patients with sporadic olivopontocerebellar atrophy (sOPCA) present various combinations of cerebellar and extracerebellar signs. However, the differential diagnosis between these two forms of cerebellar degeneration is often a challenge for the clinician. Therefore, any test helping in this differential diagnosis might have a potential clinical interest. In this study, our goal was to investigate the adaptation to increased inertia in patients with sOPCA exhibiting combined cerebellar and pyramidal signs, during the performance of fast wrist flexions. We found that these patients exhibited a hypermetria which remained unchanged after addition of inertia, because they were unable to increase neither their agonist activity (launching force), nor their antagonist activity (braking force). This contrasts with our previous findings in patients with CCA. In these latter, the hypermetria worsened when the inertial load of the hand increased because those patients were able to increase their agonist activity, but not their antagonist activity. The adaptation to inertia might thus help to differentiate CCA and sOPCA. PMID- 9349673 TI - Activity levels of a beta1,6 N-acetylglucosaminyltransferase in lymphomonocytes from multiple sclerosis patients. AB - The activity of the Golgi glycosyltransferase beta1,6 N acetylglucosaminyltransferase (core 2 GlcNAc-T), which plays a role in T-cell activation and cell-cell adhesion, appears to be modulated in resting lymphomonocytes during different phases of multiple sclerosis (MS). In particular, a significant decrease (25-30%) of the enzyme activity was observed, with respect to healthy subjects, in MS patients who were in relapse or in the very early stages of remission. A similar trend was found to be associated with patients affected by active lesions. A statistically significant decrease in the enzyme activity was also observed in patients with the progressive form. By contrast, core 2 GlcNAc-T activity did not appear correlated with duration of the disease. Interestingly, MS individuals under treatment with IFN-beta1a, an immunosuppressive agent, showed levels of activity which were comparable with those observed in healthy subjects. Together, these observations suggest that down-regulation of core 2 GlcNAc-T activity is linked to the occurrence of acute phases in the relapsing-remitting form and to the progressive form of the disease, probably caused by altered expression of glycoproteins which are involved in lymphomonocyte activation and/or interaction with the endothelium. Additionally, it appears that the enzyme assay may provide a useful marker of the disease activity and the effects of therapeutical approaches. PMID- 9349674 TI - Relation of leptin and neuropeptide Y in human blood and cerebrospinal fluid. AB - Leptin and neuropeptide Y (NPY) are involved in the regulation of food intake and body weight. Both hormones act through specific receptors in the central nervous system. The objective of this study was to investigate the relation of leptin and NPY in human plasma and cerebrospinal fluid (CSF). Leptin and NPY in CSF and in serum/plasma were measured by radioimmunoassays in 35 patients. Leptin concentrations in serum were 100-200 fold higher than in CSF. There was a significant correlation between leptin levels in CSF and in serum (r=0.88, P<0.0001). Female patients had significantly higher leptin serum concentrations than males (16.6+/-10.9 microg/l vs. 6.5+/-7.3 microg/l, P=0.002). In contrast, NPY levels were only twofold higher in CSF than in plasma. There was no relation between leptin and NPY in CSF and serum/plasma, respectively. The ratio of CSF and peripheral leptin levels did not correlate with the respective albumin ratio, indicating that leptin did not merely leak into the CSF via a defective blood-CSF barrier. It is concluded that leptin uptake from the circulation into CSF is a regulated process. The NPY concentration in CSF is not directly related to leptin CSF levels. PMID- 9349675 TI - IgM anti-sulfatide autoantibodies: patterns of binding to cerebellum, dorsal root ganglion and peripheral nerve. AB - Anti-sulfatide antibodies are associated with polyneuropathies having a prominent sensory component, but with variable degrees of motor and sensory loss, gait dysfunction and demyelination. In this study, we asked whether patterns of IgM binding to neural tissue in anti-sulfatide serums also demonstrated heterogeneity. We used immunocytochemical methods to examine IgM binding to peripheral nerve, dorsal root ganglion, and cerebellum in 41 serums with high titers of IgM anti-sulfatide antibodies. Our results showed that there were several different patterns of IgM binding to neural tissues in anti-sulfatide serums. In peripheral nerve the most common targets of IgM were axons, resident macrophages or Schwann cell cytoplasm. In the cerebellum, IgM bound to neuronal nuclei, white matter, or neuropil in molecular and granule cell layers. There was little binding of IgM to structures in the dorsal root ganglion. Patterns of IgM binding to peripheral nerve and cerebellum were related. Binding to neuronal nuclei in the cerebellum was usually found in serums that recognized peripheral nerve axons or macrophages. Serums with binding of IgM to cerebellar white matter usually recognized Schwann cell cytoplasm. We conclude that IgM anti-sulfatide antibodies may have several different tissue binding patterns in the peripheral and central nervous systems. These differences may be related to the variation in clinical neuropathy syndromes associated with apparently similar anti-sulfatide antibodies. PMID- 9349676 TI - The effect of movement frequency on cerebral activation: a positron emission tomography study. AB - Knowledge of the effect of performance frequency on activation of motor areas in positron emission tomography (PET) studies is crucial to the interpretation of experiments in which performance is a variable. We studied this effect in six normal right-handed volunteers using H2(15)O PET to measure regional cerebral blood flow (rCBF). Subjects were scanned at rest and while executing joystick movements with the right hand in freely chosen directions at different frequencies. Significant frequency dependent increases in rCBF were demonstrated in contralateral sensorimotor cortex, lateral premotor cortex bilaterally, posterior supplementary motor area (SMA), and ipsilateral cerebellar hemisphere and vermis. The striatum and the right dorsal prefrontal cortex were also activated by joystick movement compared with rest, but the magnitude of activation found in these areas was independent of the frequency of movement. The results suggest that primary motor cortex, posterior SMA, lateral premotor cortex and cerebellum are involved in determining the basic parameters of movement. Frequency dependent activation in these areas suggests phasic activity related to movement. In contrast, activation of the dorsal prefrontal cortex and the striatum is not frequency dependent. This may reflect continuous rather than phasic activity in these areas during the task and suggests their role is not simply related to movement execution but higher level during this free selection joystick task. PMID- 9349678 TI - Severe hyponatremia as poor prognostic factor in childhood neurologic diseases. AB - We determined the duration of hospital stay and neurologic sequelae in 72 patients divided into three groups according to the serum sodium level on admission. The mean duration of stay in those with normal sodium (Group I) was 18 days compared with 37 and 69 days for those with mild (Group II) and moderate to severe deficits (Group III) respectively. The differences were statistically significant (F=327.2, P<0.01). Thirty-one out of the 35 patients in Group I recovered fully; all the Group II subjects had residual deficit and all the 14 patients in Group III were either left with severe deficit (10) or died (4). Pediatric neurological cases having severe hyponatremia should be considered as a high risk group for poor outcome in spite of cautious correction. PMID- 9349677 TI - Rhythmic facilitation of gait training in hemiparetic stroke rehabilitation. AB - Experimental and control groups of 10 hemiparetic stroke patients each underwent a 6 week, twice daily gait training program. The control group participated in a conventional physical therapy gait program. The experimental group trained in the same basic program with the addition of rhythmic auditory stimulation (RAS). Patients entered the study as soon as they could complete 5 strides with hand held assistance. The training program had to be completed within 3 months of the patients' stroke. In the experimental group RAS was used as a timekeeper to synchronize step patterns and gradually entrain higher stride frequencies. Study groups were equated by gender, lesion site, and age. Motor function was assessed at pretest using Barthel, Fugl-Meyer, and Berg Scales. Walking patterns were assessed during pre- and post-test without RAS present. Pre- vs post-test measures revealed a statistically significant (P<0.05) increase in velocity (164% vs 107%), stride length (88% vs 34%), and reduction in EMG amplitude variability of the gastrocnemius muscle (69% vs 33%) for the RAS-training group compared to the control group. The difference in stride symmetry improvement (32% in the RAS group vs 16% in the control group) was statistically not significant. The data offer evidence that RAS is an efficient tool to enhance efforts in gait rehabilitation with acute stroke patients. PMID- 9349679 TI - Spatial distribution of corticospinal potentials following transcranial electric and magnetic stimulation in human spinal cord. AB - To investigate the spatial distribution of the human corticospinal tract in the spinal cord, evoked spinal cord potentials (ESCPs) following transcranial electrical and magnetic stimulation were recorded simultaneously from both the anterior and posterior epidural space in five anesthetized patients. One ESCP component following transcranial electrical stimulation (D-wave) and at least two ESCP components (initially D-wave and later I-wave) following transcranial magnetic stimulation were recorded in all subjects. The negative peak latency of all the potentials recorded from the posterior epidural space was the same as that recorded anteriorly. The amplitude ratio of the ESCP following electrical stimulation (posterior/anterior) was 1.10+/-0.12, while that of ESCPs following magnetic stimulation was 1.08+/-0.12 (N1) and 1.15+/-0.16 (N2). These results suggest that lateral corticospinal tract descending dorsolateral fasciculus in the spinal cord is main corticospinal pathway and spatial distribution of D and I waves are similar in the human cervical cord. PMID- 9349680 TI - Treatable lumbosacral polyradiculitis masquerading as diabetic amyotrophy. AB - Patients with diabetic amyotrophy may have an inflammatory vasculopathy and may obtain reversal of neurological deficits with immunosuppression. We present a patient with NIDDM, subacute onset of painful asymmetric polyradiculopathy, and unilateral enhancement of lumbar nerve roots on MRI. Clinical improvement and resolution of nerve root enhancement occurred with immunosuppression. We suggest, therefore, that nerve biopsy and gadolinum-enhanced lumbosacral MRI be performed in all patients presenting with diabetic amyotrophy. If nerve root enhancement is present or if nerve biopsy shows perivascular infiltrates, we recommend a trial of immunosuppression. PMID- 9349681 TI - Prominent sensory ataxia in Guillain-Barre syndrome associated with IgG anti-GD1b antibody. AB - Sensitization with GD1b has been shown to cause sensory neuropathy in rabbit. A patient with chronic sensory-dominant polyneuropathy who had IgM antibody specifically to GD1b has been reported previously. This report describes the first patient with acute demyelinating polyneuropathy with prominent sensory symptoms who had a high titer of serum IgG anti-GD1b antibody. The serum reacted with neither GM1 nor with other b-series gangliosides (GD2, GD3, GT1b and GQ1b). Improvement in symptoms was coincident with decrease in IgG anti-GD1b antibody titer after plasmapheresis. This case supports the experimental results in rabbit suggesting that anti-GD1b antibody functions in the development of sensory ataxia. PMID- 9349682 TI - An overview of chemoprevention: current status and future prospects. AB - The optimal way of dealing with any disease is by prevention. This is particularly true of cancer, with all of its complexities. A major problem that exists for cancer prevention is that we do not know the cause of over 50% of cancers. Even when causes are known, serious difficulties often exist in removing them. To the extent that causality cannot be dealt with effectively, other strategies merit consideration. One is chemoprevention. A great strength of chemoprevention is that a large number of compounds can prevent the occurrence of cancer, and a variety of mechanisms exist for producing such protection. Much of this review deals with efficacy, toxicity, and mechanisms of action of chemopreventive agents. Gap areas in information are discussed, as well as opportunities for producing compounds with optimal attributes. Chemoprevention is not simple, and successes may not come quickly. However, for both the general population and, even more urgently, for individuals at high risk, chemoprevention has the potential of providing an important means for cancer prevention. PMID- 9349683 TI - The role of epidemiology in cancer prevention. AB - Cancer is a major cause of morbidity and mortality throughout the world. As the population lives to an older age, cancer incidence and mortality are expected to increase because of the strong relationship between cancer and advancing age. Epidemiology plays a key role in cancer prevention and control by describing the distribution of cancer and discovering risk factors for cancer. Epidemiologic study designs include descriptive, ecologic, cross-sectional, and analytic (cohort, case-control, and intervention) studies. In the past 50 years, epidemiologic research has helped to elucidate many risk factors for cancer. Lifestyle factors such as smoking, diet, alcohol consumption, reproduction (pregnancy, lactation, age at menarche, and menopause), obesity, and inactivity have been suggested as the major contributors to the development of cancer. Epidemiologists have demonstrated that cancer is largely an avoidable disease and estimated that more than two-thirds of cancer might be prevented through lifestyle modification. Epidemiologic research is crucial to public health and cancer prevention. Individuals or communities at increased risk of cancer can be targeted for risk factor modification, as well as for secondary prevention and chemoprevention strategies. PMID- 9349684 TI - Properties of intraepithelial neoplasia relevant to cancer chemoprevention and to the development of surrogate end points for clinical trials. AB - Cancer chemoprevention is defined as the prevention of cancer by the administration of diet supplements or drugs. A drug discovery effort should therefore focus on finding agents that will avert the process of intraepithelial neoplasia which precedes invasive cancer. Over 30 agents developed by the chemoprevention program at the National Cancer Institute are being tested against intraepithelial neoplasia of many organ sites in more than 80 clinical trials. Two basic mechanisms underlie the onset and development of intraepithelial neoplasia. First is the development of the two precursor lesions of chronic diffuse epithelial hyperplasia and genomic instability, the latter being produced by "mutator" mutations in genes responsible for genomic stability, by gene copy amplification or loss from DNA breakage-fusion-anaphase-bridge cycles, by unequal sister chromatid exchange, and by accumulation of double minutes. Second is the development of multicentric intraepithelial neoplastic lesions which independently progress through each of the following processes at a continuously accelerating rate: clonal evolution, hyperproliferation, production of genomic structural variants, and apoptosis. Recommended chemoprevention strategies based on these mechanisms are (i) the development of better technology for early diagnosis, (ii) the development of multiple agents that block intralesional proliferation at steps along the signal pathway of mitotic signal transduction and along the signal pathway of synthesis of daughter cell components, (iii) the development of nontoxic anti-inflammatory agents, anitoxidants, antimutagens, and proapoptotics, (iv) the avoidance of "clonal escape" through use of drug combinations, and (v) the use of computer-assisted quantitative image analysis to assay modulation of surrogate end points in chemoprevention clinical trials. PMID- 9349685 TI - DNA damage as an intermediate biomarker in intervention studies. AB - The development of sensitive assays for measurement of DNA damage in humans has great potential for enhancing intervention studies. Methods for DNA adduct measurement include immunoassays, [32p] postlabeling, high-performance liquid chromatography with fluorescence or electrochemical detection, and gas chromatography/mass spectroscopy. It is now well established that DNA adducts are a marker of exposure to various environmental, lifestyle, or occupational chemical carcinogens. Our own studies concentrate on immunologic detection of adducts by enzyme-linked immunosorbent assay (ELISA) of isolated DNA or quantitative immunohistochemical analysis of intact cells. Polycyclic aromatic hydrocarbon (PAH)-DNA adducts are elevated in blood cells of foundry and coke oven workers, individuals with high levels of exposure to environmental air pollution, and smokers. The study in smokers also found an inverse relationship between serum antioxidants and PAH-DNA, and is the basis for an ongoing antioxidant intervention. DNA adducts of PAH and 4-aminobiphenyl and oxidative DNA damage (8-oxo-deoxyguanosine) are being measured in blood mononuclear cells and exfoliated oral and bladder cells from subjects on antioxidants or placebo. Data on published intervention studies investigating oxidative damage and general aromatic DNA adducts measured by postlabeling are also summarized. These studies have already demonstrated that DNA adducts can be modulated by interventions and suggest that they can provide important mechanistic information in support of larger scale studies. PMID- 9349686 TI - Molecular epidemiology: carcinogen-DNA adducts and genetic susceptibility. AB - Molecular epidemiological studies assess individual chemical exposures and genetic susceptibility in order to identify cancer risk. Such studies incorporate the development, application, and validation of biomarkers of cancer risk in order to enhance cancer risk assessments, focus cancer prevention strategies, and elucidate mechanisms of carcinogenesis. Current studies of molecular epidemiology are based upon an understanding of the complex, multistage process of carcinogenesis and interindividual variations in response to carcinogenic exposures. Quantitative methods to measure human exposures to carcinogens continue to improve and have been successfully applied to a number of epidemiological studies. Genetic predispositions to cancer, both inherited and acquired, have been and continue to be identified. The combined approach of associating genetic polymorphisms with carcinogen-DNA adduct measurements, in order to assess cancer risk, is showing considerable promise. It is hoped that, in the future, molecular epidemiologists will be able to develop a risk profile for an individual that includes assessment of multiple biomarkers. The field has the near-term potential to have a significant impact on regulatory quantitative risk assessments, which may aid in the determination of allowable exposures. Molecular epidemiological data may also aid in the identification of individuals who will most benefit by cancer prevention strategies. PMID- 9349687 TI - Approaches to cancer prevention based on an understanding of N-nitrosamine carcinogenesis. AB - Understanding carcinogenesis is critical for development of rational approaches to cancer prevention. This paper uses N-nitrosamine carcinogenesis as an example. N-Nitrosamines are a large group of potent carcinogens. Approximately 300 different N-nitrosamines are carcinogenic. At least 30 animal species are responsive to their effects. There is little doubt that humans exposed to sufficient amounts of N-nitrosamines would also be susceptible to their carcinogenic effects. Human exposure to preformed N-nitrosamines occurs through the diet, in certain occupational settings, and through the use of tobacco products, cosmetics, pharmaceutical products, and agricultural chemicals. Diminishing human exposure to these carcinogens is one approach to prevention of cancer, and this has been accomplished in many instances, although exposure to N nitrosamines in tobacco products is still unacceptably high. Human exposure to N nitrosamines also occurs by nitrosation of amines in the body, via their acid or bacterial catalyzed reaction with nitrite, or by reaction with products of nitric oxide generated during inflammation or infection. A second approach toward prevention of N-nitrosamine carcinogenesis is inhibition of this endogenous N nitrosamine formation. Substantial reductions have been achieved with ascorbic acid and other nitrite scavengers. N-Nitrosamines undergo a simple cytochrome P450-mediated metabolic activation step, which is critical for their carcinogenicity. The third approach involves the use of chemopreventive agents that block this step, or other steps in the carcinogenic process. A large number of potent chemopreventive agents against nitrosamine carcinogenesis have been identified. Chemoprevention of lung cancer induced by the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is discussed as an example of this approach. PMID- 9349688 TI - Detoxication enzymes and chemoprevention. AB - Detoxication enzymes protect cells from a wide variety of xenobiotics and endogenous toxins. Current data suggest that the balance between the Phase I carcinogen-activating enzymes and the Phase II detoxifying enzymes is critical to determining an individual's risk for cancer. Human deficiencies in Phase II enzyme activity, specifically glutathione-S-transferase (GST), have been identified and associated with increased risk for colon cancer. The increased frequency of the GST M1 null genotype among individuals with primarily smoking related cancers has been documented. Induction of Phase II enzymes by naturally occurring or synthetic agents represents a promising strategy for cancer prevention. Both the required characteristics of potential chemopreventive agents and the role of the antioxidant response element in the monofunctional induction of Phase II enzymes have been discussed. The synthetic dithiolthione oltipraz induces a battery of Phase II enzymes and inhibits chemically induced tumors in a variety of target organs. Its ability to induce Phase II enzymes in human colon tissue and blood lymphocytes has been reported. Other promising inducers with chemopreventive activity include the isothiocyanates and polyphenols. These data collectively support the future development of Phase II enzyme inducers as clinical chemopreventive agents. PMID- 9349689 TI - Inhibition of cyclooxygenase: a novel approach to cancer prevention. AB - An expanding body of evidence indicates that downregulation of the cyclooxygenases (Cox-1 and Cox-2) will be an important strategy for preventing cancer because cyclooxygenases catalyze the formation of prostaglandins (PGs), and PGs have multiple effects that favor tumorigenesis. PGs also are more abundant in cancers than in the normal tissues from which cancers arise. Overexpression of Cox-2 in epithelial cells inhibits apoptosis and increases the invasiveness of tumor cells; inhibitors of Cox (e.g., NSAIDS) are chemopreventive; and tumorigenesis is inhibited in Cox-2 knockout mice. We focus in this review on strategies to selectively inhibit and downregulate the Cox-2 isoform. This is important because simultaneous inhibition of Cox-1 (constitutively expressed) and Cox-2 (inducible isoform), which is achieved with classical NSAIDs, interferes with the housekeeping functions of Cox-1 and thereby causes serious side effects, such as peptic ulcer disease. Simultaneous inhibition of Cox-1 and Cox-2 hence is not a realistic approach for chemoprevention in individuals at low to moderate risk for cancer. On the other hand, it appears possible to avoid many NSAID-dependent side effects by selective inhibition of Cox-2, which is also the isoform that is upregulated in benign and malignant tumors. Through understanding the biochemistry of these enzymes and the regulation of Cox-1 and Cox-2 gene expression, we review how Cox-2 can be regulated selectively as a target for chemopreventive therapy. We also discuss the potential importance and advantages of a multifaceted approach to diminishing the function of Cox-2 (i.e., combining inhibitors of enzyme function with inhibitors of gene expression). PMID- 9349690 TI - Dietary control of cancer. AB - Many laboratory studies and human epidemiological data suggest that most cancer deaths are attributable to lifestyle, including nutritional factors and tobacco and alcohol consumption. Tobacco consumption is causally related to cancer of the lung, mouth, larynx, esophagus, bladder, kidney, and pancreas. Nutrients and non nutrient dietary components probably account for cancer of the colon, breast, prostate, and stomach. This report is based on literature and our own data pertaining to the role of dietary fat, calories, and fiber in the development of colon and breast cancer. We also discuss the evidence from epidemiological, mechanistic, and preclinical efficacy studies indicating a protective effect of micronutrients, non-nutrients, and certain antioxidants in food against oral and lung cancers. Given the continuing cancer burden and the relatively slow impact of proven cancer treatment strategies in reducing cancer mortality, it is essential to evaluate promising nutrients and non-nutrients in foods as chemopreventive agents in persons at increased risk for cancer. Development of reliable intermediate biomarkers is valuable for clinical chemoprevention intervention trials. The purpose of this report is to provide the reader with plausible approaches to cancer control. PMID- 9349691 TI - Dietary fatty acids and prevention of hormone-responsive cancer. AB - The results from some, but not all, epidemiological studies indicate that the level of dietary fat intake and the nature of the constituent fatty acids influence both breast and prostate cancer risk, and disease progression. These observations derive support from the use of animal models, which demonstrate that polyunsaturated omega-6 fatty acids stimulate mammary carcinogenesis and tumor growth and metastasis, whereas long-chain omega-3 fatty acids exhibit inhibitory effects. While studies of prostate cancer are less advanced, the available data are in agreement with those designed to evaluate the associations between breast cancer and dietary fatty acids. In both cases, a multiplicity of biological actions of eicosanoids derived from tumor cell arachidonate metabolism appear to elicit responses, both in the tumor itself and in the host cells that subscribe to its microenvironment. This review concludes that clinical intervention trials designed to reduce total fat intake and increase the ratio of omega-3 to omega-6 fatty acids in the diet should be targeted at groups at a relatively high risk for breast or prostate cancer, and also at postsurgically treated cancer patients with the objective of preventing disease recurrence. PMID- 9349692 TI - Some perspectives on dietary inhibition of carcinogenesis: studies with curcumin and tea. AB - Topical application of curcumin inhibits chemically induced carcinogenesis on mouse skin, and oral administration of curcumin inhibits chemically induced oral, forestomach, duodenal, and colon carcinogenesis. Curcumin and other inhibitors of cyclooxygenase and lipoxygenase are thought to inhibit carcinogenesis by preventing the formation of arachidonic acid metabolites. In contrast to our expectation of a tumorigenic effect of arachidonic acid, we found that treatment of 7,12-dimethylbenz[a]anthracene-initiated mouse skin with very high doses of arachidonic acid twice daily, 5 days a week for 26 weeks, failed to result in tumors. We considered the possibility that some of the cancer chemopreventive effects of curcumin may be related to an effect of this compound on cellular differentiation, and we investigated the effect of curcumin on differentiation in the human promyelocytic HL-60 leukemia cell model system. Although curcumin alone had little or no effect on cellular differentiation, when it was combined with all-trans retinoic acid or 1alpha,25-dihydroxyvitamin D3 a synergistic effect was observed. It is possible that many dietary chemicals in fruits, vegetables, and other edible plants can prevent cancer by synergizing with endogenously produced stimulators of differentiation such as all-trans retinoic acid, 1alpha,25 dihydroxyvitamin D3, and butyrate. More research is needed to test this hypothesis. Administration of green or black tea inhibits carcinogenesis in several animal models, and tumor growth is also inhibited. Several examples were presented of chemopreventive agents that inhibit carcinogenesis in one animal model but enhance carcinogenesis in a different animal model. Greater efforts should be made to understand mechanisms of cancer chemoprevention and to determine whether a potential chemopreventive agent is useful in many experimental settings or whether it is useful in only a limited number of experimental settings. PMID- 9349693 TI - Inhibition of proliferation and modulation of estradiol metabolism: novel mechanisms for breast cancer prevention by the phytochemical indole-3-carbinol. AB - Aberrant proliferation is an early-occurring intermediate event in carcinogenesis whose inhibition may represent preventive intervention. Indole-3-carbinol (I3C), a glucosinolate metabolite from cruciferous vegetables, inhibits organ site carcinogenesis in rodent models. Clinically relevant biochemical and cellular mechanisms for the anticarcinogenic effects of I3C, however, remain unclear. Experiments were conducted on reduction mammoplasty derived 184-B5 cells initiated with chemical carcinogen (184-B5/BP) or with oncogene (184-B5/HER), and on mammary-carcinoma-derived MDA-MD-231 cells to examine whether (i) I3C inhibits aberrant proliferation in initiated and transformed cells, and (ii) inhibition of aberrant proliferation is associated with altered cell-cycle progression, estradiol (E2) metabolism, and apoptosis. Aberrant proliferation in 184-B5/BP, 184-B5/HER, and MDA-MB-231 cells was evident by a 55%-67% decrease in the ratio of quiescent (Q = G0) to proliferative (P = S + M) phase of the cell cycle, a 72% 90% decrease in apoptosis, and a 76%-106% increase in anchorage-dependent growth. These cells also exhibited a 88%-90% decrease in the ratio of C2 to C16alpha hydroxylation products of E2. Treatment of 184-B5/BP, 184-B5/HER, and MDA-MB-231 cells to cytostatic dose of 50 microM I3C resulted in an 137%-210% increase in Q/P I3C ratio, a 4- to 18-fold increase in E2 metabolite ratio, a 2-fold increase in cellular apoptosis, and a 54%-61% inhibition of growth. The preventive efficacy of I3C on human mammary carcinogenesis may be due in part to its ability to regulate cell-cycle progression, increase the formation of antiproliferative E2 metabolite, and induce cellular apoptosis. PMID- 9349694 TI - The clinical evaluation of cancer prevention agents. AB - The clinical development of cancer chemoprevention agents is similar to that of cytotoxic agents. On the basis of promising preclinical studies, agents with potential efficacy are introduced to the clinic as a phase I trial to study the dose-toxicity relationship of the agent and its human pharmacology. If doses projected to have biological activity have acceptable side effects, the agent proceeds to a phase II trial, which enrolls a larger number of participants and administers the agent for a longer time period (up to 5 years). Phase IIb trials test the effect of these agents on potential biomarkers of carcinogenesis to get some indication if the agent has activity. The phase II trial also pilots study design, mechanisms of recruitment, and adherence for future phase III trials. The phase III trial of a chemopreventive agent determines its effect on cancer incidence. Close attention is also paid to long-term side effects. The unique features of cancer prevention agents and their evaluation must be considered in this stepwise clinical evaluation. These features include (i) populations recruited to prevention trials are healthy; (ii) in this population there is a low tolerance for side effects; and (iii) the clinical end point of the trial, cancer, is statistically an uncommon event. The evaluation of beta-carotene and retinol as studied in the Carotene and Retinol Efficacy Trial (CARET) is used to illustrate this stepwise clinical development. PMID- 9349695 TI - Patient participation and compliance in cancer chemoprevention trials: issues and concerns. AB - Cancer chemoprevention trials have unique characteristics that make the tasks of participant recruitment, enrollment, and long-term adherence to the study protocol and intervention regimen especially difficult. Barriers to patient accrual, long-term participation, and optimal adherence are inherent in clinical trial design and organization, and are frequently associated with the attitudes and behavioral dynamics of physicians and the participants themselves. Attracting racially and ethnically diverse populations to trial participation adds additional problems and considerations. Careful planning early in the design phase of a chemoprevention clinical trial must take into account these numerous issues. Clinical investigators should seek expert advice from a number of health care disciplines to better design chemoprevention protocols that minimize logistic complexity, maximize participant eligibility, simplify data collection, and take into account the complex behavioral dynamics of the clinical trial process. PMID- 9349696 TI - Colon cancer prevention: intervening in a multistage process. AB - Colorectal cancer, one of the most common human malignancies, is also one of the best understood. The epidemiology of this disease, and its relationship to environmental influences, particularly diet, has been extensively studied. New insights into the molecular biology and genetics of colorectal cancer also provide clues to its etiology, and high-risk populations that are most likely to benefit from preventive measures can be identified. Using this information, promising strategies for prevention of colorectal cancer are under investigation. These strategies include dietary modification, screening and adenoma removal, and antitumor agents from both natural and synthetic sources. PMID- 9349697 TI - Novel approaches to the prevention of head and neck cancer. AB - Head and neck cancer represents a paradigm for the prevention of environmentally induced diseases. Etiologic factors, including tobacco and alcohol, have been well established. Furthermore, disease development may depend not only upon such exposures but also upon a genetically defined susceptibility to exposure-induced DNA damage. As discussed in this review, the ability to identify etiologic factors, both external and host related, will lead to improved identification of individuals at risk. The development of novel strategies for cancer prevention will be promoted, among which lifestyle alteration will arguably play the most significant role. Therefore, this review places special emphasis on the translation of the understanding of disease into more effective behavioral modification strategies. PMID- 9349698 TI - Retinoids in head and neck chemoprevention. AB - Over the last few years, we have witnessed tremendous progress in both basic and clinical research on retinoids. Preclinical studies have indicated the potential of retinoids in cancer prevention and therapy, but the actual successful application of retinoids in clinical chemoprevention trials has been the recent and exciting development in the field of retinoid research. Our understanding of the role of retinoids in normal developmental processes and the differentiation of normal and malignant cells, and the fundamental discovery of the nuclear retinoid receptors that act as transcription modulating factors regulating specific gene expression have been major advances in the field of basic retinoid research. Chemoprevention is the newest research approach in our efforts to control upper-aerodigestive tract cancers, which have one of the lowest cure rates among epithelial malignancies, and in which the occurrence of second primary tumors further burdens the dismal prognosis of patients. The efficacy of retinoids in the reversal of oral premalignant lesions and the prevention of second primary tumors has generated tremendous enthusiasm among retinoid researchers, particularly those in the field of chemoprevention. Current explorations of combinations of retinoids with biologic response modifiers such as alpha-interferon, as well as new receptor-selective retinoids, hold promise for the future. PMID- 9349699 TI - Vitamin/mineral supplementation and cancer risk: international chemoprevention trials. AB - In this review, large-scale randomized intervention trials evaluating the effects of vitamin and mineral supplementation on cancer rates are summarized. The trials enrolled up to 30,000 adults who were followed for up to 12 years, and included assessments of multiple vitamin and mineral combinations in an area of China with limited micronutrient intake and one of the world's highest cancer rates; and of beta carotene, vitamin E, or selenium in several more well nourished western populations, some at very high risk of lung cancer. Results to date have been mixed. Significantly lower cancer mortality has been found among those supplemented with a combination of beta carotene, vitamin E, and selenium in the China trial and with selenium in the United States, but risks of lung cancer were increased in Finnish and American trials provided with high-dose beta carotene supplementation. In combination, the trials indicate that the relation between specific micronutrient intake and cancer risk is complex, but have provided information to target further research on the potential benefits of selenium, vitamin E, and combinations of vitamins and minerals. PMID- 9349700 TI - Differential regulation of the plasmid-encoded genes in the Shigella flexneri virulence regulon. AB - Expression of the Shigella flexneri virulence gene regulon is controlled by multiple environmental signals acting through a regulatory cascade. The primary regulator is VirF, which is a positive regulator of the secondary regulatory gene virB and the structural gene icsA. The product of the virB gene in turn activates transcription of the genes coding for the invasion proteins, and for the type III secretion system which promotes export of the invasion proteins to the bacterial cell surface. The genes making up the regulon were studied in their native locations on the 230-kb virulence plasmid. Transcriptional control was detected at each level of the regulatory cascade. A gearing effect was detected upon thermal induction of transcription in the regulon, with the virF gene being induced by about two fold, virB by 10-fold and the structural genes by 100-fold. In addition, each gene studied displayed individual characteristics in its response to stimuli such as growth medium osmolarity, pH, variations in DNA superhelicity and the presence or absence of H-NS. The primary regulatory gene virF, displayed loose regulation under standard laboratory growth conditions. Regulation was tighter at the secondary regulator virB, while control of structural gene expression was tighter still. It is proposed that this regulatory pattern ensures that energetically wasteful expression of the structural genes under inappropriate conditions is avoided while allowing the regulatory genes to be expressed sufficiently under non-permissive conditions to ensure a rapid response to inducing conditions when these arise. Once induced, fine tuning of the response can be achieved through the different sensitivities of the individual regulon members to external stimuli. PMID- 9349701 TI - Impairment of calcineurin function in Neurospora crassa reveals its essential role in hyphal growth, morphology and maintenance of the apical Ca2+ gradient. AB - The function of Neurospora crassa calcineurin was investigated in N. crassa strains transformed with a construct that provides for the inducible expression of antisense RNA for the catalytic subunit of calcineurin (cna-1). Induction of antisense RNA expression was associated with reduced levels of cna-1 mRNA and of immunodetectable CNA1 protein and decreased calcineurin enzyme activity, indicating that a conditional reduction of the target function had been achieved in antisense transformants with multiple construct integrations. Induction conditions caused growth arrest which indicated that the cna-1 gene is essential for growth of N. crassa. Growth arrest was preceded by an increase in hyphal branching, changes in hyphal morphology and concomitant loss of the distinctive tip-high Ca2+ gradient typical for growing wild-type hyphae. This demonstrates a novel and specific role for calcineurin in the precise regulation of apical growth, a common form of cellular proliferation. In vitro inhibition of N. crassa calcineurin by the complex of cyclosporin A (CsA) and cyclophilin20, and increased sensitivity of the induced transformants to the calcineurin-specific drugs CsA and FK506 imply that the drugs act in N. crassa, as in T-cells and Saccharomyces cerevisiae, by inactivating calcineurin. The finding that exposure of growing wild-type mycelium to these drugs leads to a phenotype very similar to that of the cna-1 antisense mutants is consistent with this idea. PMID- 9349702 TI - Structural characterization of the gene and corresponding cDNA for the cytochrome P450rm from Rhodotorula minuta which catalyzes formation of isobutene and 4 hydroxylation of benzoate. AB - Cytochrome P450rm was previously isolated from the basidiomycete yeast Rhodotorula minuta as a bifunctional enzyme with isobutene-forming and benzoate 4 hydroxylase activities. We cloned the gene and corresponding cDNA for P450rm in order to characterize the enzyme in the context of fungal phylogeny and physiology. From the cDNA sequence, P450rm was deduced to have 527 amino acids with a calculated molecular weight of 59136. P450rm shared 48% amino acid sequence identity with CYP53A1 from Aspergillus niger, indicating that the gene belongs to a novel subfamily of CYP53, CYP53B. However, the organization of the P450rm gene, which has eight exons and seven introns, differed completely to that of CYP53A1. Northern analysis demonstrated that the level of P450rm mRNA expression increased when L-phenylalanine was used as sole carbon source. These results suggest that P450rm has been well conserved during the evolution of fungi as a benzoate 4-hydroxylase in the dissimilation pathway starting from L phenylalanine PMID- 9349703 TI - Map-based cloning of the tomato genomic region that spans the Sw-5 tospovirus resistance gene in tomato. AB - Two yeast artificial chromosomes (YACs) containing genomic DNA from tomato have been isolated using CT220, an RFLP marker which is tightly linked to the tomato spotted wilt virus resistance gene, Sw-5. High-resolution mapping of the YAC ends and internal YAC probes demonstrated that one of the YAC clones, TY257 (400 kb), spans Sw-5. By chromosome walking in a cosmid library, the position of Sw-5 has been delimited within the YAC to a maximal chromosomal segment of 100 kb, spanned by nine overlapping cosmid clones. PMID- 9349704 TI - Genetic diversity and vegetative compatibility among Trichoderma harzianum isolates. AB - Trichoderma harzianum is the collective name of a set of asexual fungal strains which exhibit heterogeneity in genome structure, DNA sequence and behavior. Contour-clamped homogeneous field (CHEF) electrophoresis of the chromosomes of ten isolates of T. harzianum revealed six clearly distinct electrophoretic karyotypes. Of the ten isolates analyzed, four (GH12, G109, Y and YF) could be classified in a single group with identical karyotypes, while the strains T35 and 315 formed a second group. The genome size characteristic of the different isolates fell into a broad range varying from 29.6 to 56.1 Mb. Gene assignments to the resolved chromosomes showed that all genes analyzed were localized on equivalent chromosomes in the isolates belonging to the same group. Analysis of randomly amplified polymorphic DNAs from the ten isolates confirmed the classification into groups and allowed us to distinguish between isolates T35 and 315, as well as between isolates GH12, G109, Y and YF. Direct confrontation assays using isolates of the same group showed compatible interactions, whereas the same experiment carried out with isolates of different groups showed an incompatible interaction characterized by an area of cell damage. Microscopic observation of the compatible interactions showed hyphal fusions between the isolates, similar to those described for vegetative compatible groups in other fungi. The molecular karyotypes correlated well with the compatibility of the isolates. In addition, we have evaluated both electrophoretic karyotype and randomly amplified polymorphic DNAs analysis as criteria for grouping isolates within the genus according to their capacity for biocontrol of plant pathogens. PMID- 9349705 TI - Regulation of the yeast HIS7 gene by the global transcription factor Abf1p. AB - The HIS7 gene of Saccharomyces cerevisiae encodes a bifunctional glutamine amidotransferase: cyclase that catalyzes the formation of biosynthetic precursors for histidine and adenine. HIS7 is activated by Gcn4p upon amino acid starvation and by the Bas1/2p complex in response to adenine limitation. Mutation analysis of the HIS7 promoter in a deltagcn4 background revealed a polyd(A/T) stretch and a d(CT) repeat as essential elements for Gcn4p-independent basal HIS7 transcription. The protein binding this element was enriched and identified as the multifunctional DNA-binding protein Abf1p. Abf1p binds specifically to the d(CT) repeat sequence, which represents a novel Abf1p-binding motif, and protects 17 nucleotides from digestion by DNase I. In addition, Abf1p binding causes bending of the HIS7 promoter structure. PMID- 9349706 TI - High rates of polymorphism and recombination at the Opaque-2 locus in cultivated maize. AB - As a first step in the study of the functional consequences of molecular polymorphism at the Opaque-2 locus in maize, 1933 bp of the gene were sequenced in 21 inbred lines representative of the progenitors of cultivated varieties. High levels of polymorphism were found: 109 sites were variable in non-coding regions and 159 in protein-coding regions. Among the latter, 103 were nonsynonymous, resulting in 94 amino acid replacements in a 422-residue peptide. Moreover, 26 insertion/deletion polymorphisms were found, eight of them in coding regions. The high rate of polymorphism observed could indicate that the effective size of the ancestral population was very large, and/or that the O2 gene evolved under the influence of a high mutation rate or was subjected to some kind of balancing selection. The distribution of polymorphism within the sequence was not uniform. Silent polymorphisms were relatively frequent in exon 1 and rare in intron 5, whereas nonsynonymous polymorphisms were infrequent in the part of the sequence encoding the active domain of the transcription factor. The peptide sequence also showed a relatively higher level of resistance to amino acid replacements in this region. Sites in linkage disequilibrium were arranged mainly in three spatial patterns. This mosaic pattern can be related with an apparently large number of recombination events which might be a characteristic feature of this particular sample and/or imply that the O2 locus is a hot spot for recombination. PMID- 9349707 TI - Novel, developmentally specific control of Ds transposition in maize. AB - Plants form their gametes late in somatic development and, as a result, often pass somatic mutations on to their progeny. Classic examples of this process are the germinal revertants of unstable, Ac/Ds transposon-induced kernel mutations in maize: frequent and early reversion events during somatic development are generally correlated with a high frequency of revertant gametes. We have characterized a Ds allele of the maize waxy (wx) gene, wx-m5:CS7, for which the correlation between somatic and germinal reversion frequencies no longer holds. The ability of wx-m5:CS7 (CS7) to produce revertant gametes is suppressed approximately 100-fold in comparison with a second Ds allele, wx-m5:CS8 (CS8), which has an identical insertion at Wx and the same frequent and early somatic reversion pattern in endosperm. The excision of Ds from wx is not reduced 100 fold in the somatic tissues of CS7 plants as compared with CS8 plants. Suppressed formation of CS7 revertant gametes is independent of the Ac transposase source and is heritably passed to the embryos of progeny kernels; however, frequent and early somatic reversion is observed again in endosperms of these progeny kernels. This suppression appears to be caused by a dominant mutation in a trans-acting product that can suppress the germinal reversion of other Ds-induced alleles as well; the mutation is tightly linked to Wx but is not in the CS7 Ds itself. Taken together, the data suggest a novel mode of developmental control of Ac/Ds elements by the host plant, suppressing element excision in the shoot meristem. PMID- 9349708 TI - What is a bona fide mating-type gene? Internuclear complementation of mat mutants in Podospora anserina. AB - In the heterothallic ascomycete Podospora anserina, the mating-type locus is occupied by two mutually exclusive sequences termed mat+ and mat-. The mat+ sequence contains only one gene, FPR1, while the mat- sequence contains three genes: FMR1, SMR1 and SMR2. Previous studies have demonstrated that FPR1 and FMR1 are required for fertilization. Further analyses have led to the hypothesis that mat+ and mat- genes establish a mat+ and mat- nuclear identity, allowing recognition between nuclei of opposite mating type within the syncytial cells formed after fertilization. This hypothesis was based on the phenotypes of strains bearing mutations in ectopic mat genes. Here we present an analysis of mutations in resident mat- genes which suggests that, unlike FMR1 and SMR2, SMR1 is not involved in establishing nuclear identity. In fact, mutations in these two genes impair nuclear recognition, leading to uniparental progeny, while mutations in SMR1 block the sexual process, probably at a step after nuclear recognition. The nuclear identity hypothesis has also been tested through internuclear complementation tests. In these experiments, the mat- mutants were crossed with a mat+ strain carrying the wild-type mat- genes. Our rationale was that internuclear complementation should not be possible for nuclear identity genes: the relevant genes should show nucleus-restricted expression, and diffusion of their products to other nuclei should not occur. This test confirmed that SMR1 is not a bona fide mat gene since it can fulfill its function whatever its location, in either a mat- or a mat+ nucleus, and even when present in both nuclei. SMR2, but not FMR1, behaves like a nuclear identity gene with respect to internuclear complementation tests. A model is proposed that tentatively explains the ambiguous behaviour of the FMR1 gene and clarifies the respective functions of the three mat- proteins. PMID- 9349709 TI - Molecular characterization of the glnA gene encoding glutamine synthetase from the edible mushroom Agaricus bisporus. AB - The gene encoding glutamine synthetase (glnA) was isolated from an Agaricus bisporus H39 recombinant lambda phage library. The deduced A. bisporus glutamine synthetase amino acid sequence contains 354 residues. The amino acid sequence is very similar to that derived from the gene coding for glutamine synthetase of the yeast Saccharomyces cerevisiae. The open reading frame is interrupted by four introns. Northern analysis revealed that transcription of the gene is repressed upon addition of ammonium to the culture but the repression was not as strong as that of the gene encoding NADP+ -dependent glutamate dehydrogenase (gdhA). Enzyme activities are low in the presence of ammonium, glutamine and albumin and do not correlate with the mRNA levels revealed by Northern analysis. This suggests that glutamine synthetase expression in A. bisporus is also post-transcriptionally regulated by the nitrogen source. PMID- 9349710 TI - A complex glutathione transferase gene family in the housefly Musca domestica. AB - In most metazoans, the glutathione S-transferases (GST) are encoded by gene families, and are used to detoxify xenobiotics. We describe the structure of genomic loci coding for the GSTs in the housefly that have been implicated, by both genetic and biochemical means, in mediating insecticide resistance. In earlier work, we showed that one of the theta-class enzymes, MdGST-3, is overproduced in resistant flies and degrades certain insecticides. We used a fragment from a cDNA clone of MdGST-3 as a probe to screen a housefly genomic DNA bank in phage lambda. This probe detected multiple gst loci. Genes for GSTs were found in five different, nonoverlapping lambda clones, three of which carry multiple, closely linked gsts. Multiple genes for both MdGST-3 and MdGST-4 were found; some of which have introns in their 5' untranslated regions. In adults, the only MdGST-3 enzymes that are expressed are encoded by the intron-free genes. A new theta-class GST (called MdGST-5) was also discovered. Fusion genes comprising 5' MdGST-3 sequences and either MdGST-4 or MdGST-5 sequences in their 3' halves were encountered at three separate loci. The genes described here are found in both the ancestral sensitive strain and the insecticide-resistant strains. PMID- 9349711 TI - Isolation of the transposable element hupfer from the entomopathogenic fungus Beauveria bassiana by insertion mutagenesis of the nitrate reductase structural gene. AB - A transposable element has been isolated from the entomopathogenic fungus Beauveria bassiana by trapping it in the nitrate reductase structural gene, which has been cloned from this species. The element had inserted in the first exon of the nia gene and appeared to have duplicated the sequence TA at the site of insertion. It was 3336 bp long with 30-bp imperfect, inverted, terminal repeats. The element, called hupfer, contained an open reading frame encoding a 321-amino acid protein similar to the IS630- or mariner-Tcl-like transposases, and a residual sequence of about 2 kb which was not significantly similar to any published sequence. There are fewer than five copies of this transposable element present per genome in the fungus. PMID- 9349712 TI - Characterization of Streptomyces nogalater genes encoding enzymes involved in glycosylation steps in nogalamycin biosynthesis. AB - The sno gene cluster in Streptomyces nogalater ATCC 27451 contains the nogalamycin biosynthesis genes. A set of plasmid constructions carrying fragments of the sno cluster that lie downstream of snoD were used to complement the S. galilaeus mutant H039, which is blocked in rhodosamine and 2-deoxyfucose biosynthesis in the aclacinomycin pathway. Sequence analysis of this cluster revealed three contiguous open reading frames (ORFs) that were designated snoF, snoG, and snoH. Only those plasmid constructs that expressed SnoG were able to complement H039. SnoG shows similarity to GalE, a UDP-glucose-4-epimerase catalyzing the epimerization of UDP-glucose to UDP-galactose. The putative SnoF protein is similar to 3,5-epimerases involved in rhamnose biosynthesis. The deduced product of snoH is a 489-amino acid polypeptide. It is similar to the product of dau ORF3 found in the daunomycin cluster. However its function is still unclear. Based on the complementation experiments and sequence analysis, this part of the sno cluster is suggested to be involved in the biosynthesis of the sugar portion of nogalamycin. Interestingly, SnoA, a transcriptional activator for the sno minimal polyketide synthase, is also needed to express this cluster. PMID- 9349713 TI - An Arabidopsis gene encoding a putative 14-3-3-interacting protein, caffeic acid/5-hydroxyferulic acid O-methyltransferase. AB - AFT1, a 14-3-3 protein from Arabidopsis thaliana, was used as a 'bait' in the two hybrid system to identify its interacting proteins. A caffeic acid/5 hydroxyferulic acid O-methyltransferase, OMT1, was identified as one of the several proteins that specifically interacts with AFT1 in yeast cells. The physical interaction between AFT1 and a partial OMT1 polypeptide can be demonstrated in vitro by using bacterially expressed proteins. A single copy gene was found to encode OMT1 in Arabidopsis, and its expression is both spatially and temporally regulated. PMID- 9349714 TI - A novel levansucrase-levanase gene cluster in Bacillus stearothermophilus ATCC12980. AB - A 3.1 kb fragment of Bacillus stearothermophilus ATCC12980 DNA permitted growth of Escherichia coli on sucrose. The fragment encoded genes for a levansucrase (surB) and also a levanase (surC). SurB and SurC shared 97% and 43% amino acid identity with the corresponding SacB and SacC proteins of Bacillus subtilis, whose sacB and sacC genes are organised very differently. PMID- 9349715 TI - Cloning and sequencing of a human cDNA encoding ornithine decarboxylase antizyme inhibitor. AB - We report here cloning and sequencing human antizyme inhibitor from a human kidney cDNA library. Amino acid sequence deduced from the nucleotide sequence shows 92.9% identity to that of rat antizyme inhibitor. Northern blot analysis reveals that antizyme inhibitor is expressed in human liver. PMID- 9349716 TI - Cloning of the phytases from Emericella nidulans and the thermophilic fungus Talaromyces thermophilus. AB - Phytases (EC 3.1.3.8) belong to the family of histidine acid phosphatases. We have cloned the phytases of the fungi Emericella nidulans and Talaromyces thermophilus. The putative enzyme encoded by the E. nidulans sequence consists of 463 amino acids and has a Mr of 51785. The protein deduced from the T. thermophilus sequence consists of 466 amino acids corresponding to a Mr of 51450. Both predicted amino acid sequences exhibited high identity (48% to 67%) to known phytases. This high level of identity allowed the modelling of all available fungal phytases based on the three-dimensional structure coordinates of the Aspergillus niger phytase. By this approach we identified 21 amino acids which are conserved in fungal phyA phytases and are part of the residues forming the substrate pocket. Furthermore, potential glycosylation sites were identified and compared between the aforementioned phytases and the A. niger phytase. PMID- 9349717 TI - Molecular cloning of a novel alternatively spliced nuclear protein. AB - Using the yeast two hybrid system, we isolated a rat cDNA (E3-3) coding for a new protein with no homology to any other protein in the database. E3-3 is ubiquitously expressed. Variants that most likely arise through alternative splicing encode truncated forms of the protein. Testis is the only tissue that predominantly expresses the longest protein variant. When this variant is tagged with enhanced green fluorescent protein, the protein is located in the nucleus. PMID- 9349718 TI - Characterisation of the human CD30 ligand gene structure. AB - The gene for the human CD30 ligand was molecularly cloned, sequenced and characterised. The gene structure consisted of four exons and three intervening introns spaced over 17.1 kb of genomic DNA. The 5' flanking region of the CD30L gene was sequenced and analysis of the region revealed the presence of several regulatory regions including a poly-dT element directly upstream from the transcription start site and consensus sequences for AP4, IK2, MZF1, E47 and ELK/cETS1. The absence of a canionical TATA motif suggests CD30L gene expression is regulated by a TATA-less promoter. PMID- 9349719 TI - Cloning and characterization of the 5' flanking region of human ATP-citrate lyase gene. AB - Two phage clones, lambda hgACL21 and lambda hgACL28, harboring the 5' flanking region of human ATP-citrate lyase (ACL) gene were identified by screening about 1.5 X 10(6) recombinant plaques from the lambdaEMBL3-human placental genomic DNA library. The 5' flanking region of ACL had the CAAT box on -92 bp from the transcription initiation site (+1), however, the TATA box was not found. The primer extension and rapid amplification of cDNA end showed that mRNA is transcribed at a thymine extending 12 bp upstream of the reported cDNA end. The sequences of 5' flanking region in 1.5 kb size of human ACL showed 60% homology with those of rat; however, no homology was found in the exon 1 and intron 1 region. Several consensus sequences, including four Sp1 binding sites, were found in the 5' flanking region of this gene. The promoter activity was assayed by transfecting the 3' or 5' deletion clones of ACL-chloramphenicol acetyl transferase (CAT) plasmid into PLC/PRF5 cells. The clone that contains the part of the first intron sequences from -659 to +440 bp showed the highest CAT activity in the transient transfection assay. High promoter activities were maintained until the transcription initiation site was removed. It is suggested that the sequences from -213 to +12 which contain three Sp1-binding sequences, CAAT box, and the transcription initiation site were necessary as a mean of for exerting the basal promoter activity of ACL gene. PMID- 9349720 TI - Antisense S-oligodeoxynucleotides down-regulate TGFbeta-production by Kupffer cells from CCl4-injured rat livers. AB - TGFbeta is a pleiotropic cytokine involved in multiple physiological and pathophysiological regulatory mechanisms. Since TGFbeta is a disparate modulator of cell recruitment, proliferation and extracellular matrix phenotype for mesenchymal and nonmesenchymal cells, we have been investigating the role of this cytokine in the pathophysiology of liver. In the present paper we investigate which hepatic cell types from CCl4-injured rat livers express TGFbeta mRNA and produce TGFbeta in culture, with the aim of further obliterating its biological activity by means of antisense technology. We performed a series of comprehensive molecular studies of in situ hybridization, northern blots, and RT-PCR and we found that only non-parenchymal cells produce TGFbeta while its expression in hepatocytes was absent. Consistent with the in situ hybridization findings, we observed that Kupffer cells expressed high steady-state levels of TGFbeta mRNA, while circulating monocytes expressed a smaller amount of TGFbeta transcripts. We did not detect TGFbeta gene expression in endothelial cells. These findings were further confirmed by RT-PCR analyses. TGFbeta activity, as measured by inhibition of [3H]thymidine incorporation by Mv 1 Lu mink lung epithelial cells, was down regulated in culture by antisense phosphorothioate oligonucleotides. These effects of antisense oligomers were dose-dependent and the sense oligonucleotides had no effect at the same concentration. PMID- 9349721 TI - Cloning, sequencing, and expression of dnaK-operon proteins from the thermophilic bacterium Thermus thermophilus. AB - We propose that the dnaK operon of Thermus thermophilus HB8 is composed of three functionally linked genes: dnaK, grpE, and dnaJ. The dnaK and dnaJ gene products are most closely related to their cyanobacterial homologs. The DnaK protein sequence places T. thermophilus in the plastid Hsp70 subfamily. In contrast, the grpE translated sequence is most similar to GrpE from Clostridium acetobutylicum, a Gram-positive anaerobic bacterium. A single promoter region, with homology to the Escherichia coli consensus promoter sequences recognized by the sigma70 and sigma32 transcription factors, precedes the postulated operon. This promoter is heat-shock inducible. The dnaK mRNA level increased more than 30 times upon 10 min of heat shock (from 70 degrees C to 85 degrees C). A strong transcription terminating sequence was found between the dnaK and grpE genes. The individual genes were cloned into pET expression vectors and the thermophilic proteins were overproduced at high levels in E. coli and purified to homogeneity. The recombinant T. thermophilus DnaK protein was shown to have a weak ATP-hydrolytic activity, with an optimum at 90 degrees C. The ATPase was stimulated by the presence of GrpE and DnaJ. Another open reading frame, coding for ClpB heat-shock protein, was found downstream of the dnaK operon. PMID- 9349722 TI - Molecular cloning of rat leukemia inhibitory factor receptor alpha-chain gene and its expression during pregnancy. AB - Leukemia inhibitory factor (LIF) is a secreted glycoprotein and a pluripotent growth factor that acts on diverse cell systems. LIF transmits its effects via binding to transmembrane receptors, of which both high- and low-affinity forms have been identified. In this study, we analyzed the structure and expression of rat LIF receptor alpha-chain (rLIFR alpha) cDNA. A full-length clone of the cDNA encoding the membrane-bound form of rLIFR alpha protein was prepared by a combination of LA-PCR and 5' RACE using DNA reverse-transcribed from total RNA isolated from the livers of day-12 and day-14 pregnant rats as templates. The nucleotide sequence of a full-length clone was determined and further confirmed by analysis of shorter DNA fragment prepared by PCR using pfu polymerase. The gene for rLIFR alpha encodes a 1093 amino acid residue protein. The rLIFR alpha protein shows a high degree of similarity to mouse and human LIF receptor alpha chain protein (89% and 76% amino acid sequence identities, respectively). Only one molecular species of mRNA for the rLIFR alpha gene was detected in the liver and placenta. rLIFR alpha was expressed in liver of both non-pregnant and pregnant rats. The level of mRNA for the rLIFR alpha gene in placenta was maximum on day 16 of pregnancy. PMID- 9349723 TI - Molecular cloning and expression of rat kallistatin gene. AB - We have previously purified and cloned human kallistatin and rat kallikrein binding protein (RKBP), which are tissue kallikrein inhibitors belonging to the serine proteinase inhibitor superfamily. In this study, we have cloned and sequenced the gene encoding rat kallistatin with Phe-Phe-Ser-Ala-Gln at positions P2-P3', which is identical to the reactive center of human kallistatin. Rat kallistatin is highly similar to human kallistatin, sharing 68% and 57% sequence identity at the cDNA and the amino acid levels. The rat kallistatin gene exists in a single copy and is located on chromosome 6. An SphI RFLP is found between SHR and WKY rats at or near the rat kallistatin gene locus. Two amino acid polymorphisms of the rat kallistatin gene between these two strains were found by sequence analysis. A candidate promoter in the 5'-flanking region (109 bp) of the rat kallistatin gene has been identified by reporter assays. The expression of rat kallistatin in the liver is growth-dependent and down-regulated during acute phase inflammation. Recombinant rat kallistatin produced in E. coli is able to bind to tissue kallikrein, and the interaction is inhibited by heparin. These characteristics define rat kallistatin as the counterpart of human kallistatin. PMID- 9349725 TI - Heat shock-induced excessive relaxation of DNA in Escherichia coli mutants lacking the histone-like protein HU. AB - Plasmid DNA in exponentially growing Escherichia coli immediately relaxes after heat shock but the relaxed DNA re-supercoils rapidly, the despite continued presence of the heat shock conditions. We have now obtained genetic evidence indicating that the histone-like protein HU of E. coli is required for this re supercoiling of DNA. Plasmid DNA in a hupA-hupB double gene-disruption mutant relaxed excessively after heat shock, while the relaxation of DNA in a himA-himD double gene-disruption mutant and in an hns insertion mutant was transient, thereby indicating that HU protein, but not IHF or H-NS proteins, is required for the re-supercoiling of DNA. Exposure of the hupA-hupB double mutant to a temperature of 50 degrees C led to both excessive relaxation of DNA and to a decrease in viable cell number but temperatures lower than 46 degrees C did not lead to these events. Based on these results, we propose that HU protein maintains the negative supercoiling of DNA during thermal stress and contributes to cellular thermotolerance in E. coli. PMID- 9349724 TI - Multiple splice variants of phosphodiesterase PDE4C cloned from human lung and testis. AB - Four closely related cyclic-nucleotide specific phosphodiesterase (PDE4) genes have been identified in both humans and rats: PDE4A, 4B, 4C and 4D. We have now cloned cDNAs for multiple splice variants of human PDE4C. Two splice variants, PDE4C-791 and PDE4C-426, were isolated from a fetal lung library. The longest open reading frame (ORF) of 791 amino acids (aa) is encoded by PDE4C-791, which is similar to a recently described cDNA [Engels, P., Sullivan, M., Muller, T. and Lubbert, H. FEBS Lett. 358 (1995) 305-10], except that an alternative 5'-end sequence upstream of the first methionine extends the PDE4C-791 ORF by 79 aa. The PDE4C-426 variant contains 3 insertions that are located 5' to the catalytic domain and encode several in-frame stop codons. The predicted 426 aa protein initiates at a methionine 365 aa within PDE4C-791. A baculovirus clone starting at this methionine expressed an enzymatically active protein. Two additional splice variants, PDE4C-delta54 and PDE4C-delta109, were found in testis mRNA. PDE4C-delta54 contained a novel 5'-end region and a deletion of 162 nt; the predicted protein deletes 54 aa from the amino-terminal region. The PDE4C-delta54 protein produced in baculovirus-infected cells was enzymatically active and sensitive to PDE4-specific inhibitors. The PDE4C-delta109 protein is similar to PDE4C-delta54 but has an additional 55 aa deleted in the catalytic domain; it lacked enzymatic activity. Analysis of uncloned total mRNA from 4 tissue sources by polymerase chain reaction (PCR) confirmed the presence of mRNAs with the two deletions and three insertions that we observed in cDNA clones. The PDE4C-delta54 variant was found only in testis and the 5'-extended region of PDE4C-791 was seen only in lung and the melanoma cell line G361. Hence, tissue-specific expression of various PDE4C isoforms should be considered in understanding how these gene products modulate cellular responses to cAMP. PMID- 9349726 TI - Molecular characterization of the cyclin-dependent kinase inhibitor p27 promoter. AB - p27Kip1 is a member of the family of cyclin-dependent kinase inhibitors (CKIs), which play critical roles in the regulation of cell cycle. To study the transcriptional regulation that controls the expression of p27, we have isolated the p27 promoter, defined its transcription initiation site, and performed various analyses for sequences upstream to 3 kb. Transient transfection assays using fusion reporters containing progressively truncated p27 promoter fragments showed that a region of 170 bp upstream of the start site is sufficient for maximal transcription activity. Detailed sequence analysis of this 170 bp region identified several GC-rich segments, putative sites of the transcription factor Sp1. Footprinting experiments revealed two Sp1-protected boxes, named BoxI and BoxII, which are located at positions -133 to -117 and -87 to -72, respectively. Binding of Sp1 to the two boxes was further demonstrated by gel mobility shift assays and supershift assays. Co-transfection studies in Drosophila Schneider line 2 cells showed that Sp1 indeed activates the p27 promoter constructs that harbor one or both of the GC-rich sequences. Furthermore, the GC-rich sequences could confer Sp1-dependent transactivation to a heterologous prolactin minimal promoter. Mutations in the GC-rich sequences abolished both binding and transactivation by Sp1. Taken together, our data strongly show that the p27 promoter is activated by the ubiquitously expressed transcription factor Sp1, which may provide a molecular mechanism for the constitutive nature of p27 transcription. PMID- 9349727 TI - Sarcoplasmic reticulum-sarcolemma interactions and vascular smooth muscle tone. AB - A characteristic of vascular smooth muscle cell morphology is a close apposition of its peripheral sarcoplasmic reticulum (SR) with the sarcolomma; this arrangement gives rise to important functional interactions whereby the peripheral SR regulates Ca2+ influx and vascular tone. We review here the key evidence supporting the following aspects of SR-sarcolemma interactions while establishing a conceptual framework encompassing (i) the SR ultrastructure and functions, (ii) the integration of the sarcolemmal Na+-Ca2+ exchanger and the peripheral SR in the mediation of a bidirectional Ca2+ exchange between the peripheral SR and the extracellular space, (iii) the existence of a higher myoplasmic free Ca2+ concentration [Ca2+]myo in the subsarcolemmal space formed between the sarcolemma and the peripheral SR relative to the [Ca2+]myo of the inner myoplasm in the resting smooth muscle cell, (iv) the division of the subsarcolemmal space into functional microdomains, (v) the existence of spontaneous localized bursts of Ca2+ release from the peripheral SR (Ca2+ sparks) towards the sarcolemma, (vi) the physiological triggering of nonlocalized Ca2+ release from the peripheral SR by Ca2+ influx (Ca2+-induced Ca2+ release), and (vii) capacitative Ca2+ entry in vascular smooth muscle. We present an overview of the physiological and pathological implications of these interactions. PMID- 9349728 TI - Lack of bradykinin-induced smooth muscle cell hyperpolarization despite heterocellular dye coupling and endothelial cell hyperpolarization in porcine ciliary artery. AB - In porcine coronary artery, bradykinin-induced endothelium-dependent vasodilatations are associated with simultaneous endothelium as well as endothelium-dependent smooth muscle cell (SMC) hyperpolarizations. In contrast, in porcine ciliary artery bradykinin evokes endothelium-dependent relaxations, but no change in SMC membrane potential. This study addresses the question of whether the lack of bradykinin-induced SMC hyperpolarization is also associated with an absence of endothelial hyperpolarization in porcine ciliary artery. With a microelectrode to impale cells in arterial strips, a 12-mV transient bradykinin induced hyperpolarization was measured in endothelial cells. Bradykinin evoked no SMC hyperpolarization deep in the media. Only occasionally, a slight 4-mV hyperpolarization could be recorded in some SMC next to the endothelium. The endothelial intracellular injection (through the recording electrode) of the fluorescent tracers, lucifer yellow or ethidium bromide, showed the existence of a heterocellular dye coupling between endothelial cells and SMC. These observations in porcine ciliary artery demonstrate that the lack of bradykinin induced endothelium-dependent SMC hyperpolarization is not due to an absence of endothelial cell hyperpolarization, but most likely to an insufficient electrotonic propagation space constant from endothelial cells to SMC, despite the presence of a dye coupling between these cells. PMID- 9349729 TI - Projections of sympathetic non-noradrenergic neurons to skeletal muscle arteries in guinea-pig limbs vary with the metabolic character of muscles. AB - This study set out to examine in detail the distribution of axons of sympathetic non-noradrenergic neurons innervating the arterial bed in skeletal muscles of the forelimb and hindlimb of guinea-pigs. The distribution of non-noradrenergic axons with immunoreactivity to vasoactive intestinal peptide (VIP) was examined in limb muscles of different histochemical character. The immunohistochemical demonstration of myosin heavy chain from fast-twitch muscle, and the histochemical demonstration of adenosine triphosphatase and succinic dehydrogenase, were used to determine the muscle fibre profile of 6 different limb muscles. Muscles included the oxidative type I muscle fibre-rich accessory semimembranosus muscle, the predominantly glycolytic type II muscle fibre-rich cranial gracilis and biceps brachii muscles and the plantaris, gastrocnemius medial head and triceps brachii long head of mixed muscle fibre composition. The frequency with which the VIP-immunoreactive (VIP-IR) axons innervated intramuscular arterial vessels was compared between categories of muscles defined by their muscle fibre profile. This study demonstrated that the projection of non noradrenergic sympathetic neurons to skeletal muscle vasculature was widespread in guinea-pig limb muscles, but that it was not uniform. VIP-IR axons were more likely to innervate the arterial vasculature of muscles with a high proportion of type I and/or oxidative muscle fibres than of muscles with a large proportion of type IIb muscle fibres. This relationship between the distribution of sympathetic non-noradrenergic axons and the metabolic characteristics of muscle suggests that these presumed vasodilator neurons have an important role in matching blood flow to the particular metabolic demands of different limb muscles. PMID- 9349731 TI - Induction of smooth muscle cell nitric oxide synthase by human leukaemia inhibitory factor: effects in vitro and in vivo. AB - We have previously shown that human leukaemia inhibitory factor (hLIF) inhibits perivascular cuff-induced neointimal formation in the rabbit carotid artery. Since nitric oxide (NO) is a known inhibitor of smooth muscle growth, NO synthase (NOS) activity in the presence of hLIF was examined in vivo and in vitro. In rabbit aortic smooth muscle cell (SMC) culture, significant NOS activity was observed at 50 pg/ml hLIF, with maximal activity at 5 ng/ml. In the presence of the NOS inhibitor L-NAME, hLIF-induced activation of NOS was greatly decreased, however it was still 63-fold higher than in control (p < 0.05). SMC-DNA synthesis was significantly reduced (-47%) following incubation with hLIF plus L-arginine, the substrate required for NO production (p < 0.05), with no effect observed in the absence of L-arginine. Silastic cuff placement over the right carotid artery of rabbits resulted in a neointima 19.3 +/- 5.4% of total wall cross-sectional area, which was increased in the presence of L-NAME (27.0 +/- 2.0%; p < 0.05) and reduced in the presence of L-arginine (11.3 +/- 2.0%; p < 0.05). The effect of L arginine was ameliorated by co-administration of L-NAME (16.4 +/- 1.5%). However, administration of L-NAME with hLIF had no effect on the potent inhibition of neointimal formation by hLIF (3.2 +/- 2.5 vs. 2.1 +/- 5.4%, respectively). Similarly, with hLIF administration, NOS activity in the cuffed carotid increased to 269.0 +/- 14.0% of saline-treated controls and remained significantly higher with co-administration of L-NAME (188.5 +/- 14.7%). These results indicate that hLIF causes superinduction of NO by SMC, and that it is, either partially or wholly, through this mechanism that hLIF is a potent inhibitor of neointimal formation in vivo and of smooth muscle proliferation in vitro. PMID- 9349730 TI - Subendothelial proteoglycan synthesis and transforming growth factor beta distribution correlate with susceptibility to atherosclerosis. AB - Coronary bypass vessels, saphenous vein (SV) and internal thoracic artery (ITA), differ in susceptibility to atherosclerosis and medium- to long-term patency. Whereas most ITA remain patent (90% at 10 years), 20% of SV grafts fail in the first year and approximately 45% fail within 10 years. Reasons for these differences are not fully understood. Loss of SV patency may reflect early metabolic events, particularly increased proteoglycan (PG) synthesis which contributes to intimal volume and promotes atherogenesis through retention of atherogenic lipoproteins. We determined, in vitro, the PG metabolic activity of SV, ITA, and human coronary arteries through autoradiographic detection of incorporated [3H]glucosamine. SV had significantly higher levels of PG synthesis than ITA, especially in the subendothelial zone and after time (7 days) in culture. Patterns of synthesis in coronary vessels were similar to SV with high levels of incorporation in the subendothelial zone of thickened intima (> 100 microm). Increased subendothelial labelling in SV was due to increased PG synthesis, not decreased degradation. ITA showed no propensity for upregulation of subendothelial PG synthesis. Immunohistochemistry showed TGF-beta1 and TGF beta2 localised primarily to the subendothelial zone of SV and coronary arteries. With time in culture immunostaining increased in parallel with increased PG synthesis. Subendothelial TGF-beta1 and TGF-beta2 were absent in ITA. A panspecific TGF-beta neutralising antibody reduced subendothelial PG synthesis in SV and coronary arteries by 50 and 60%, respectively. These results support the idea that vessels susceptible to atherosclerosis show increased accumulation of subendothelial PG mediated by TGF-beta. PMID- 9349732 TI - Aortic calcification produced by vitamin D3 plus nicotine. AB - Calcification of the elastic arteries of the young rat by treatment with vitamin D and nicotine (VDN) has been proposed as an animal model of arterial calcification associated with age and age-related vascular pathology in man. The calcium-binding protein, S-100, which is found in human atherosclerotic lesions was associated with medial calcification of the aorta in VDN rats, especially in cases of severe calcification. Calcification (total calcium content: 366 +/- 87, n = 12 in VDN vs. 24 +/- 2 micromol g(-1) aortic dry weight in controls, n = 13) involved elastocalcinosis leading to elastolysis as revealed by a fall in the amount of desmosine and isodesmosine in the aortic wall (266 +/- 17 and 254 +/- 15 in VDN vs. 655 +/- 56 and 588 +/- 30 microg g(-1) aortic dry weight in controls). The decrease in elastin was associated with an increase in the stiffness of the aortic wall (elastic modulus: 15.1 +/- 1.8 in VDN vs. 6.7 +/- 0.5 10(6) dyn cm(-2) in controls), an increase in end-systolic stress (central systolic aortic pressure: 152 +/- 6 in VDN vs. 136 +/- 2 mm Hg in controls) (at a normotensive mean pressure level) and left ventricular hypertrophy (heart weight/body weight 2.51 +/- 0.10 in VDN vs. 2.24 +/- 0.07 g kg(-1) in controls). In conclusion, the mechanisms and consequences of aortic calcification in VDN show several similarities with calcification occurring in human athero- and arteriosclerosis. PMID- 9349733 TI - Cold storage induces an endothelium-independent relaxation to hypoxia/reoxygenation in porcine coronary arteries. AB - Tissues are often cold stored for physiological studies and for clinical transplantation. We report that cold storage induces a relaxation to reoxygenation after hypoxia (H/R) in de-endothelialized porcine coronary arteries. In fresh denuded arteries stimulated with U46619, H/R did not elicit relaxation. However, after overnight cold storage (4 degrees C), H/R elicited a transient relaxation with peak relaxation of 56 +/- 8% (n = 8), which was reproducible after 2 days of cold storage. The H/R relaxation was inhibited by methylene blue (10 microM) and LY83583 (10 microM), O2-hemoglobin (1 microM), or N(G)-methyl-L-arginine (0.2 mM), but neither N(G)-nitro-L-arginine (0.2 mM) nor cyclo-oxygenase inhibition was effective. Importantly, the H/R relaxation was attenuated by KCl (40 mM) or tetrabutylammonium chloride (5 mM), a non-selective inhibitor of K+ channels. Interestingly, authentic nitric oxide (NO)- or S nitroso-N-acetylpenicillamine (SNAP)-induced relaxations were enhanced by cold storage in U46619 (0.1 microM) contractures. When tissues were contracted with KCl (40 mM), the enhancement in NO- or SNAP-induced relaxation by cold storage was markedly smaller than with U46619. Neither catalase (1,200 U/ml) nor 3-amino triazole (50 mM), an inhibitor of catalase, affected the H/R relaxation. The duration of H/R relaxation also increased with the period of incubation at 37 degrees C in the organ bath. This was blocked by inhibition of NO synthesis or guanylate cyclase. Moreover, inhibition of protein synthesis with actinomycin D (0.1 microM) and cycloheximide (10 microM), or dexamethasone (1 microM), an inhibitor of NO synthase induction, blocked this increase in the duration of the H/R relaxation. The results suggest that in smooth muscle induction of NO pathway relaxation, which is in part mediated by K+ channels and inducible NO synthase, may be of importance to the understanding of ischemia/reperfusion responses in cold-stored arteries. PMID- 9349744 TI - Stress, 11beta-HSD, and Leydig cell function. PMID- 9349745 TI - Genomic imprinting: gametic mechanisms and somatic consequences. PMID- 9349746 TI - Prevalent decrease of the EGF content in the periurethral zone of BPH tissue induced by treatment with finasteride or flutamide. AB - The aim of the present investigation is to verify whether treatment with Finasteride or Flutamide influences the regional distribution of testosterone (T), dihydrotestosterone (DHT), and epidermal growth factor (EGF) in benign prostatic hyperplasia (BPH) tissue. Thirty seven BPH patients were studied: 15 untreated, 9 treated with Flutamide (750 mg/day for 2 months), and 13 treated with Finasteride (5 mg/day for 3 months). Testosterone and DHT were evaluated by radioimmunoassay (RIA) after purification of the extracts on celite columns, and EGF was evaluated by RIA after purification on Sep-pak C18 cartridges in total tissue and in periurethral, subcapsular, and intermediate zone. In the untreated group, T, DHT, and EGF of the periurethral region are higher than those of the subcapsular zone (P < 0.01 for T and P < 0.001 for DHT and EGF). In the Flutamide group, DHT is not modified, T is increased (P = 0.045), and EGF is decreased in total tissue (P < 0.02) and in the periurethral zone (P < 0.01). In the Finasteride group, T is increased (P < 0.001), and DHT and EGF are decreased (P < 0.001), particularly in the periurethral zone. A positive linear correlation between DHT and EGF is observed in the Finasteride and in the untreated groups. In conclusion, in BPH the production of EGF is a DHT-dependent receptor-mediated function. The reduction of this growth factor during both treatments, associated with a fall of DHT in only the Finasteride group, is particularly evident in the periurethral zone. Since Finasteride reduces prostatic volume, mainly of the periurethral zone, we can speculate that DHT is responsible, either directly or indirectly through growth factors such as EGF for the enlargement of this region and thus responsible for urinary obstruction. PMID- 9349747 TI - Serum androgens: associations with prostate cancer risk and hair patterning. AB - Cancer of the prostate is the leading cancer among American men, yet few risk factors have been established. Hair growth and development are influenced by androgens, and it has long been suspected that prostate cancer also is responsive to these hormones. A blinded, case-control study was undertaken to determine if hair patterning is associated with risk of prostate cancer, as well as specific hormonal profiles. The study accrued 315 male subjects who were stratified with regard to age, race, and case-control status (159 prostate cancer cases/156 controls). Hair-patterning classification and serum levels of total and free testosterone (T), sex hormone binding globulin, and dihydrotestosterone (DHT) were performed. Data indicate that hair patterning did not differ between prostate cancer cases and controls; however, significant hormonal differences were detected between the two groups. Free T was greater among cases than in controls (16.4 +/- 6.1 vs. 14.9 +/- 4.8 pg/ml, P = 0.02). Conversely, DHT-related ratios were greater among controls (P = 0.03 for DHT/T and P = 0.01 for DHT/free T). Several strong associations also were found between hormone levels and hair patterning. Men with vertex and frontal baldness had higher levels of free T (16.5 +/- 5.5 and 16.2 +/- 8.0 pg/ml, respectively) when compared to men with either little or no hair loss (14.8 +/- 4.7 pg/ml) (P = 0.01). Data suggest that increased levels of free T may be a risk factor for prostatic carcinoma. In addition, although no differences in hair patterning were detected between cases and controls within this older population, further research (i.e., prospective trials or case-control studies among younger men) may be necessary to determine if hair patterning serves as a viable biomarker for this disease, especially given the strong association between free T levels and baldness. PMID- 9349748 TI - Stability and transcriptional regulation of gamma-glutamyl transpeptidase mRNA expression in the initial segment of the rat epididymis. AB - Gamma-glutamyl transpeptidase (GGT) is an enzyme believed to play a role in the protection of maturing spermatozoa in the epididymis. Our previous studies have shown that four GGT mRNAs (I-IV) transcribed from the single-copy rat GGT gene are differentially expressed and regulated in the rat epididymis. In particular, the normal expression of GGT mRNA(IV) in the epididymal initial segment is dependent upon the presence of testicular factors. The objective of this study was to test the hypothesis that the decreased expression of GGT mRNA(IV) in the initial segment following the in vivo removal of testicular factors by efferent duct ligation (EDL) is due to a decrease in stability and/or transcription rate. The stability of the GGT mRNAs was evaluated by measuring the rate of mRNA decay. These stability studies showed that GGT mRNA(IV) exhibited a rapid initial decay that slowed at later times to a decay rate similar to that of GGT mRNAs(II,III). The decay rates were not different following sham-operation or EDL, and thus the stability of GGT mRNAs were not influenced by the in vivo loss of testicular factors. Results of transcription analysis revealed that the transcription rate of GGT mRNA(IV) in the initial segment fell by approximately 68% following a 12 hour EDL. Additionally, secondary-structure models indicate two families of folding patterns for GGT mRNA(IV), which could be the reason for the two decay regimes detected in the stability study. Thus, the decreased expression level of GGT mRNA(IV) in the initial segment following the in vivo loss of testicular factors is a function of a decreased transcription rate and intricate decay kinetics. PMID- 9349750 TI - Relationship between sleep-related erections and testosterone levels in men. AB - In order to identify a possible threshold for a serum testosterone level below which sleep-related erections are impaired and to compare this threshold with the normal laboratory range of testosterone serum levels, we studied 201 men, including hypogonadal and eugonadal subjects. The protocol included nocturnal penile tumescence and rigidity monitoring and the assay of basal testosterone, prolactin, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) serum levels. The subjects were assigned to eight groups according to their testosterone serum levels. Group 1 had testosterone between 0 ng/dl and 99 ng/dl; the following seven groups had testosterone levels increased by 100 ng/dl per group. The groups of subjects with higher testosterone serum levels showed almost constantly higher values for the erectile parameters we studied than the subjects with serum testosterone < or = 99 ng/dl. On the contrary, subjects with higher testosterone serum levels showed higher values for only some erectile parameters compared to the subjects with serum testosterone between 100 and 199 ng/dl, without any significant difference among the groups with testosterone serum levels in the normal range. Our data suggest that the serum testosterone threshold for sleep-related erections is lower than the low end of the normal laboratory male range and is about 200 ng/dl. Further efforts are needed to find the precise serum testosterone ranges related to normal sleep-related erections and to normal sexual behavior, the testosterone ranges of which will probably not coincide. PMID- 9349749 TI - Comparison of responses to adrenomedullin and calcitonin gene-related peptide in the feline erection model. AB - The purpose of the present study was to investigate the effects of intracavernosal injections of adrenomedullin (ADM) and calcitonin gene-related peptide (CGRP), two structurally similar peptides, on penile erection in the anesthetized cat. Erectile responses to ADM and CGRP were compared with responses to a standard drug combination (1.65 mg papaverine, 25 microg phentolamine, and 0.5 microg prostaglandin E1 [PGE1]). Intracavernosal injections of ADM (0.1-3 nmol) and CGRP (0.01-0.3 nmol) induced erection in a dose-dependent manner. The maximal increase in intracavernosal pressure in response to ADM was a 75% increase, while the maximal response to CGRP was comparable to that induced by the reference combination, and the maximal increase in penile length was comparable with ADM, CGRP, and the standard drug combination. The duration of the maximal pressure increase and the total duration of the response to ADM and CGRP were more abbreviated than with the control combination, and systemic blood pressure was reduced significantly after administration of CGRP, the control combination, and the higher doses of ADM. The nitric oxide synthase inhibitor, L NAME, and the K+(ATP)-channel antagonist, glybenclamide, had no effect on the erectile response to CGRP or ADM. The CGRP receptor antagonist CGRP(8-37) attenuated the erectile response to CGRP but not to ADM. These data suggest that the erectile responses to ADM and CGRP are not mediated by nitric oxide release or the opening of K+(ATP) channels, two mechanisms reported to be involved in penile erection, and that CGRP and ADM induce penile erection by activating different receptors. PMID- 9349751 TI - The cellular mechanisms of corticotropin-releasing hormone (CRH)-stimulated steroidogenesis in mouse Leydig cells are similar to those for LH. AB - Previous reports have demonstrated that corticotropin-releasing hormone (CRH) treatment of primary cultures of mouse Leydig cells and MA-10 mouse Leydig tumor cells results in a dose-dependent stimulation of steroidogenesis, probably by acting through the cAMP/protein kinase A second messenger pathway. Based on this observation, the mechanism of CRH-stimulated steroidogenesis is now further investigated and compared to trophic hormone stimulation. Both cell types were treated with human chorionic gonadotropin (hCG) or CRH in the absence and presence of the following agents: the translation inhibitor cycloheximide, the transcription inhibitor actinomycin D, the protonophore carbonyl cyanide m chlorophenylhydrozone (mCCCP), which disrupts the mitochondrial electrochemical gradient or the phorbol ester, phorbol-12-myristate 13-acetate (PMA), a stimulator of protein kinase C. Cortico-releasing hormone-stimulated steroidogenesis was completely blocked by cycloheximide in both cell types, indicating that CRH-stimulated steroidogenesis in mouse Leydig cells requires ongoing protein synthesis. Actinomycin D had profound inhibitory effects on CRH stimulated steroidogenesis in MA-10 cells, and this inhibition was greater than that seen in mouse primary Leydig cells. mCCCP severely inhibited CRH-stimulated steroid production in both cell types, indicating that an electrochemical gradient across the inner mitochondrial membrane is required for CRH-stimulated steroidogenesis. In addition, PMA inhibited hCG- and CRH-stimulated steroidogenesis in MA-10 cells and CRH-stimulated steroidogenesis in primary Leydig cells, suggesting that activation of the protein kinase C pathway can influence protein kinase A stimulated steroidogenesis. Results of these studies suggest that the mouse Leydig cell steroidogenic response to CRH shares many similarities to that of the LH response. PMID- 9349752 TI - Mild hypothermia induces apoptosis in rat testis at specific stages of the seminiferous epithelium. AB - This study was designed to determine the effects of short episodes of mild hypothermia on cell apoptosis in the testis of the adult rat. Apoptosis was assessed by in situ DNA 3'-end labeling in the testes from rats killed 2 hours after cooling (10 degrees C for 30 minutes), quantitated, and compared with spontaneous apoptosis found in control animals. A significantly increased number of dying germ cells appeared after cold treatment at specific stages (stage XIV, P < 0.0001; stage XII, P < 0.001) of the seminiferous epithelium, mainly due to dying primary metaphasic spermatocytes (P < 0.0001), followed by secondary interphase spermatocytes (P < 0.001), and A2 spermatogonia (P < 0.05). The highly specific effect of mild hypothermia on germ cell apoptosis suggests that the process is tightly regulated. PMID- 9349753 TI - Partial purification and localization of platelet-activating factor acetylhydrolase from bovine seminal plasma. AB - Platelet-activating factor (PAF) is a potent lipid mediator that is inactivated by platelet-activating factor acetylhydrolase (PAF-AH). Platelet-activating factor bioactivity has been detected in bovine sperm phospholipids and PAF-AH activity is extraordinarily high in bovine seminal plasma. The purpose of this study was to purify and characterize partially the PAF-AH in bovine seminal plasma. Platelet-activating factor acetylhydrolase was partially purified from bovine seminal plasma using gelatin-agarose and ion-exchange chromatography and nondenaturing polyacrylamide gel electrophoresis (PAGE). Enzyme activity was increased 11-fold over seminal plasma with a yield of 11%. Platelet-activating factor acetylhydrolase activity was eluted from a single band with a R(f) of 0.258 from a nondenaturing preparative PAGE gel along with several other proteins of varying molecular weights. Following separation by sodium dodecyl sulfate (SDS)-PAGE under reducing conditions, PAF-AH was identified as a approximately 60 kD band by western blotting using antiserum directed against human blood PAF-AH. N-terminal sequencing of the approximately 60 kD band, followed by amino acid sequence similarity searching, demonstrated a single-sequence match with PAF-AH from bovine blood. Based on western blotting, a approximately 60-kD band corresponding to PAF-AH was detected in seminal vesicle fluid but not in samples of washed, sonicated sperm or sperm plasma membranes where activity was low (<5% and <0.3%, respectively, of that in seminal plasma), suggesting that seminal plasma PAF-AH does not bind tightly to sperm. Specific PAF-AH activity measured in seminal vesicle fluid was in the lower range of that in seminal plasma. These results demonstrate that PAF-AH activity in bovine seminal plasma is due to PAF AH secreted by the seminal vesicles with sequence homology to the enzyme in human blood. PMID- 9349754 TI - Chromatin structural changes in sperm after scrotal insulation of Holstein bulls. AB - The reported effects on semen quality ascribed to testicular heat stress generally relate to traits impacting sperm transport and fertilizing ability but not to the genetic material contained by the sperm. To characterize the effects of testicular heat stress on sperm chromatin, susceptibility of DNA in sperm nuclear chromatin to in situ acid denaturation was measured by flow cytometry after staining with acridine orange using the sperm chromatin structure assay (SCSA). Semen was collected from Holstein bulls at 3-day intervals, before and after 48-hour scrotal insulation, until the morphologically abnormal sperm content in raw semen exceeded 50%. After cryopreservation in egg yolk-citrate extender, semen was thawed and sampled during incubation in vitro at 38.5 degrees C. Overall, SCSA results showed that chromatin susceptibility to denaturation was increased for sperm collected post- vs. preinsulation and was more pronounced for sperm presumably in the testes during insulation than for those sperm presumably in the epididymides. Increased susceptibility was detected as early as the first collection postinsulation; however, chromatin of sperm presumably in the proximal epididymis during insulation did not appear to have been detrimentally affected. Chromatin susceptibility to denaturation increased with increased incubation time in vitro, but the rate of change in susceptibility during incubation did not differ among pre- vs. postinsulation specimens. We conclude that elevated scrotal temperatures adversely affect both epididymal and testicular sperm by reducing sperm chromatin stability. The effects of heat stress on the chromatin of epididymal sperm were more subtle than those exhibited by testicular sperm but detectable within close proximity to the heat stress event. PMID- 9349755 TI - Time course of spontaneous in vitro sperm acrosome reaction. AB - The acrosome reaction (AR) is an exocytotic process essential for sperm penetration of the zona pellucida and binding to the oocyte (DeJonge, 1994). Evaluation of in vitro AR can suggest fertility potential. The purpose of this study was to determine AR as a function of time after removal of sperm from the seminal fluid using a novel test, the Acrobeads test (Fertility Technologies, Natick, Massachusetts), which uses paramagnetic beads coated with MH61, a monoclonal antibody that binds to acrosome-reacted sperm. Specimens were acquired from known fertile donors (n = 9) and in vitro fertilization (IVF) patients (n = 8) with no apparent male factor on the day of the IVF and processed by a minipercoll wash at 30 minutes after ejaculation. An aliquot of washed sperm was then divided into two portions. The first was placed with the Acrobeads (according to the manufacturer's instructions) and assessed for bead binding after 6 and 24 hours with the beads. The second aliquot of washed sperm was held at room temperature for 24 hours, then exposed to the beads, with bead binding assessed at 6 and 24 hours later (30 and 48 hours after washing). The Acrobeads score was determined by assessing the binding of MH61 beads in each of four replicates with a resulting score of 1 (lowest) to 4 (highest). The mean (+/-SD) motility was 62.0% (7.5) at 6 hours, 52.3% (6.4) at 24 hours, 55.9% (10.4) at 30 hours, and 54.7% (8.4) at 48 hours after removal from the seminal fluid. At 6 hours after washing and exposure to the beads, the score was 0.077 (0.27) with a range of 0-1; one donor specimen gave a score of 1, while all others had a score of 0. At 24 hours postremoval from the semen, donor and patient sperm were positive for the test, with a mean score of 3.6 (0.65). The mean fertilization rate for the IVF patients was 64.4% (range 33-90). When sperm were held for 24 hours prior to the test, there was little or no bead binding 6 hours later (score of 0.46 +/- 0.77) and at 24 hours later (48 hours after washing) (mean score of 0.25 +/- 0.45). These data suggest that completion of the acrosome reaction occurs by 24 hours after removal of the sperm from the seminal fluid. Since the MH61 beads bind to specific residues on the inner acrosomal membrane, these data also suggest that following the acrosome reaction at 24 hours with removal of the outer acrosomal contents and acrosomal matrix, the inner acrosomal membrane may be modified in some way that does not allow MH61 beads to bind to the sperm. PMID- 9349756 TI - Surgery, science, and respiratory failure. PMID- 9349757 TI - Bronchogenic cysts and esophageal duplications: common origins and treatment. AB - BACKGROUND/PURPOSE: Bronchogenic cysts and esophageal duplications are usually considered as separate foregut malformations. Yet, both are thought to arise from the same embryological event, division of the embryonic foregut, and they share common histological characteristics, often making their clinical differentiation difficult. METHODS: A retrospective review of the cases of 68 children treated at a single institution between 1937 and 1995 was performed. Thirty children were girls (44%) and 38 were boys (56%). Ages ranged from newborn to 24 years. Complete records were available in all children. Fourteen of these 68 children were asymptomatic. RESULTS: Respiratory (54%) or gastrointestinal (13%) symptoms were the most frequent presenting problems. The majority of children were treated by resection of the cyst (52 of 68; 76%), while 9 of 68 (13%) required lobectomy for intraparenchymal lesions. Three children underwent marsupialization, with all of these children requiring additional surgery for recurrent disease. Five children (5 of 68; 7%) had multiple cysts. The mortality rate from this series was 10% (7 of 68). Two deaths were caused by perioperative exsanguination, one related to bleeding from a cyst lined with gastric mucosa with subsequent ulceration and hemorrhage into the esophagus. Two deaths occurred secondary to septic complications, one from an esophageal leak and the other from an intraparenchymal abscess. Two deaths were caused by respiratory failure; one was unrelated (SIDS). The majority of cysts found on histological review were lined by respiratory epithelium or bronchial glands (51 of 68; 75%). Gastrointestinal epithelium was present in cysts of nine children, only two of which were clinically diagnosed as esophageal duplications. Twenty-one cases (21 of 68; 31%) were classified as esophageal duplications based on the intramural location of the cyst, yet 15 of 21 (71%) contained respiratory epithelium, substantiating the hypothesis of the common origin of these lesions. CONCLUSIONS: The histological similarity and anatomic proximity of the "bronchogenic cysts" and the intramural "esophageal duplications" supports their common origin. The possible complications of bleeding, ulceration, infection, and obstruction of the esophagus or airway, should generally lead to prompt resection. PMID- 9349758 TI - Iatrogenic hepatic rupture in the newborn and its management by pack tamponade. AB - BACKGROUND/PURPOSE: This report compares the ultrastructure of the newborn and adult liver and emphasizes that the newborn liver is very prone to iatrogenic rupture resulting in a high morbidity and mortality. This study describes the methods used to treat this condition and suggests that pack tamponade may be the method of choice to control hemorrhage. METHODS: latrogenic liver rupture with blood loss greater than 35% estimated blood volume occurred in seven patients. Cause of rupture included perihepatic dissection (left lobe mobilization [n = 2], capsule breached surgically [n = 1]), and retraction (n = 3) or prosthetic silo manipulation (n = 1). RESULTS: Initial attempts to control the hemorrhage were unsuccessful in six of seven patients. The only secure method to obtain long-term control of the bleeding was perihepatic pack tamponade. Control was incomplete in one patient who had an associated coagulopathy. Transfusion-induced clotting defects were present in four cases. Pack removal at 24 to 96 hours was successful in five of six patients where the bleeding was stopped, the patient fully stabilized and coagulopathy was corrected. Pack removal caused renewed bleeding in one patient, and repacking was unsuccessful. Ultrastructural differences between newborn and adult livers were investigated. The newborn liver contains little fibrous stroma and has a very thin capsule. Suture hepatorraphy therefore is an inappropriate technique in most instances and contraindicated if a coagulopathy is present. Surface coagulation and pack tamponade may be the only options available. A single patient in this series survived this complication. CONCLUSIONS: This review documents the serious nature of iatrogenic liver injury. Blood loss must be strictly limited by obtaining immediate control of the hemorrhage. If surface control is unsuccessful, pack tamponade should be used. Suturing the newborn liver (especially premature) produces unpredictable results. These observations suggest that pack tamponade is an effective method of controlling bleeding from the liver surface. PMID- 9349759 TI - Abnormalities of neuropeptides and neural markers in the esophagus of fetal rats with adriamycin-induced esophageal atresia. AB - BACKGROUND/PURPOSE: To investigate the distribution of neural markers and neuropeptides in esophageal atresia (EA). METHODS: A fetal rat model with Adriamycin-induced EA was used. The animals were divided into four groups: (1) control group, (2) saline-injected group, (3) Adriamycin administered but without the development of EA, and (4) Adriamycin-induced EA group. Specimens of the distal esophagus from each group were immunostained using antibodies to S100, protein gene product 9.5 (PGP), somatostatin, vasoactive intestine peptide (VIP), bombesin, galanin, substance P, neuropeptide Y (NPY), calcitonin gene-related product (CGRP), met-encephalin, nitric oxide synthase, and tyrosine hydroxylase. RESULTS: The total cross-sectional area of the distal atretic esophagus was significantly smaller than controls (P = .01), the submucosa being the component most affected (0.0465 v 0.0234 mm). Immunoreactivity for S100 and galanin were significantly elevated in the atresia group (0.0288 v 0.0079 and .001 v 0.000). In addition, there was also an increase in CGRP and Substance P in the atretic group. CONCLUSION: The elevated levels of S100 and galanin could explain the disordered motility observed in patients who had esophageal atresia. PMID- 9349760 TI - Intestinal permeability to small- and large-molecular-weight substances in the newborn rabbit. AB - BACKGROUND/PURPOSE: The authors have previously reported the occurrence of spontaneous bacterial translocation (BT) and its resolution with age in the newborn rabbit. They have also reported a close correlation between small bowel bacterial colonization (BC-SB) and BT at 1 week of age, suggesting that the presence of luminal bacteria and their production of endotoxins may increase the intestinal permeability. The aim of this study was to evaluate intestinal permeability to small and large molecules in the newborn rabbit and to correlate it with BT. MATERIALS AND METHODS: New Zealand White rabbits (n = 96) 1, 7, 14, 21, and over 120 days (adult) of age were given either C14-labeled ethylene diamine tetraacetic acid (EDTA) (MW 290) or C14-Dextran (MW 70,000) via an orogastric tube at 1 mCi per 100 g of body weight. Five hours later, blood, urine, liver, and intestine were collected, and scintillation counting was performed after solubilization. In a separate series of rabbits (n = 136), the incidence of BT, BC-SB, and small intestinal surface area (SA) were measured. RESULTS: Total permeability to Dextran decreased with age and was significantly reduced at 14 days of age. In contrast, total permeability to EDTA increased and was maximal in 7- to 14-day-old rabbits and began to decrease at 21 days of age. The incidence of BC-SB rapidly increased at 7 days of age and reached 100% at 14 days of age. The incidence of BT peaked at 7 days of life (30%) and then decreased with age. SA increased rapidly in the first 3 weeks and SA growth rate of 21-day-old rabbits was almost 1,400% compared with 1-day-old rabbits. CONCLUSIONS: This study has shown an age-related reduction of intestinal permeability to large (Dextran) and small (EDTA) molecular weight particles. However, intestinal permeability to EDTA had a different pattern than Dextran, suggesting that there may be different mechanisms of intestinal permeability to different size molecules. Intestinal permeability to EDTA closely correlated with bacterial colonization and bacterial translocation, suggesting that changes in the intestinal bacterial environment may affect the intestinal permeability, possibly by activating the immune system secondary to increases in endotoxins and bacteria. PMID- 9349761 TI - Pearls and perils in the management of prolonged, peculiar, penetrating esophageal foreign bodies in children. AB - BACKGROUND/PURPOSE: Most retained esophageal foreign bodies (FB) are identified soon after ingestion and are easily extracted. A minority of FB ingestions are not identified for weeks to years and present significant problems for retrieval. The purpose of this study was to describe the diagnostic and therapeutic strategies needed to care for children who have chronic esophageal FBs. METHODS: Five children were identified as having retained esophageal FBs 2 months to 2 years after ingestion. During the same 3-year period, 100 children who had acute FBs were identified and had their foreign bodies removed endoscopically. The average age of the children was 3 years (range, 2.4 to 3.5). RESULTS: The average age of the five children identified in this study was 3 years. The items ingested included coins, a heart pendant, a clothespin spring, and a toy soldier. Complications from chronically retained foreign bodies were bronchoesophageal fistula, mediastinitis, esophageal diverticulum, and lobar atelectasis. One patient died from an aortoesophageal fistula. In all children, endoscopic removal was attempted. Barium esophagram was then performed, and foreign bodies were eventually removed via right thoracotomy. CONCLUSIONS: Long-retained esophageal FBs are extremely morbid and life threatening. History most often identifies excess salivation, new onset asthma, and/or recurrent upper respiratory infections. Three diagnostic adjuncts are helpful in identifying the presence of a long retained FB: (1) Chest x-ray (PA and lateral), (2) barium swallow, and (3) esophagoscopy. Indications for thoracotomy for removal of foreign body include (1) Poor endoscopic visualization of FB because of inflammatory tissue and (2) Herald bleeding during endoscopy. PMID- 9349762 TI - Sequential endoscopic/laparoscopic management of cholelithiasis and choledocholithiasis in children who have sickle cell disease. AB - BACKGROUND/PURPOSE: Cholelithiasis and choledocholithiasis are common complications of sickle cell disease (SCD). With the recent advances in laparoscopic cholecystectomy (LC), which has been used successfully for the management of cholelithiasis in children who have SCD, exclusion of choledocholithiasis before LC is of great importance. METHODS: Eighteen children who had SCD, cholelithiasis, and choledocholithiasis were treated at our hospital. Seven were treated with open cholecystectomy (OC) and common bile duct (CBD) exploration, and two were treated with transduodenal sphincteroplasty. The remaining 11 patients underwent endoscopic retrograde cholangiopancreatography (ERCP), sphincterotomy, and stone extraction followed by laparoscopic cholecystectomy (LC). RESULTS: A dilated CBD noted on ultrasound, elevated alkaline phosphatase, elevated total bilirubin of more than 5 mg/dL, history of pancreatitis, either singly or in combination, should raise suspicion of choledocholithiasis, and these patients together with those who have choledocholithiasis detected on ultrasound should undergo ERCP to confirm and extract the stones before LC. CONCLUSION: This sequential approach of endoscopic sphincterotomy and stone extraction followed by LC is a safe and effective approach for the management of cholelithiasis and choledocholithiasis in children who have SCD. PMID- 9349763 TI - Magnetic resonance imaging of anal sphincters after reconstruction of high or intermediate anorectal anomalies with posterior sagittal anorectoplasty and fistula-preserving technique. AB - BACKGROUND/PURPOSE: Internal anal sphincter (IAS) function can be expected in approximately 75% of cases of high or intermediate anorectal anomaly reconstruction if the fistula region is preserved and transposed to become the new anal canal. METHODS: To investigate the morphology of the IAS structure, magnetic resonance (MR) imaging was performed postoperatively in 14 patients operated on with posterior sagittal anorectoplasty (PSARP) and fistula-preserving technique. The results were compared with the appearance of the anal canal in seven normal children. In addition, comparison was made with the images of five patients operated on with earlier pull-through techniques, in which the fistula region was resected. RESULTS: In all patients operated on with PSARP and fistula preserving technique, the MRI displayed an IAS-like smooth muscle structure encircling a closed anal canal. In comparison with normal controls, the image of this IAS was more irregular and had greater variations in thickness in different directions. Moreover, the area of the IAS structure was larger in comparison with the controls. Eleven of the 14 patients showed a positive rectoanal inhibition reflex in rectoanal manometry. However, the MR findings of the three cases lacking the reflex were not different compared with the rest of the group. The five patients operated on with earlier techniques demonstrated an open anal canal without a measurable IAS smooth muscle component. CONCLUSIONS: An IAS smooth muscle structure was seen by MRI in all patients operated on with PSARP and fistula-preserving technique independently of the severity of the malformation and the postoperative physiological IAS function. However, this structure was in most cases more voluminous and irregular compared with normal controls. PMID- 9349764 TI - Long-term anal sphincter performance after surgery for Hirschsprung's disease. AB - BACKGROUND/PURPOSE: The aim of the study was to assess anal sphincter performance in relation to clinical fecal continence in adult patients who have Hirschsprung's disease. METHODS: Fifty-four adult patients (mean age, 29 +/- 7.2 years; 46 men; 8 women) who had undergone surgery for Hirschsprung's disease during their childhood underwent anorectal manometry and clinical examination. Fecal continence was evaluated with a quantitative scoring method (scoring, 0-14; 14, normal bowel function; 10-13, good continence, no social problems; 5-9, fair continence, marked social limitations; 0-4, total incontinence). Thirty healthy adults were used as controls. RESULTS: Fourteen patients had normal bowel habits according to the quantitative scoring. The median anal resting pressure of these patients was 25 cm H2O (range, 15-37.5). The median resting pressure of patients with good continence (n = 30; median, 20 cm/H2O; range, 5-27.5) and with fair continence (n = 6; median, 15 cm/H2O; range, 5-27.5) was significantly lower (P < .01) than in patients who had normal continence. There was no statistical difference in maximal squeeze pressure between the patient groups (median normal, 52.5; good, 45; fair, 52.5). In the controls, the median resting pressure (61.5 cm H2O; range, 34-105) and maximal squeeze pressure (86 cm H2O; range, 55-148) were significantly higher than in all patient groups (P < .0001). The voluntary sphincter force (maximal squeeze pressure minus resting pressure) was similar in patients and controls (patients median, 27 cm H2O; controls median, 16 cm H2O, NS). CONCLUSIONS: There is a positive correlation between functional outcome and anal resting pressure in adults who have repaired Hirschsprung's disease. The overall low resting pressure reflects internal sphincter dysfunction, which may be caused by operative trauma. Despite this, most patients have a satisfactory functional outcome, which is probably related to normal voluntary sphincter performance. PMID- 9349765 TI - Fetal ovarian cyst decompression to prevent torsion. AB - BACKGROUND/PURPOSE: Neonates who have ovarian torsion caused by an ovarian cyst often lose their ovary because the torsion and infarction occurred antenatally. Because ultrasound scan has been so effective in diagnosing ovarian cysts in utero, we have a better understanding of their natural history and can select appropriate cases for cyst decompression in utero to prevent torsion. The authors reviewed experience with seven fetuses who had fetal ovarian cyst. METHODS: During a 26-month period, seven patients were referred for the evaluation of fetal ovarian cyst. The mean gestational age at presentation was 31.9 +/- 3.6 weeks (+/-SD; range, 27 to 37 weeks). There was no history of maternal risk factors such as diabetes mellitus or fetal risk factors such as hyperthyroidism or placentomegally. All seven cases involved isolated unilateral cysts without associated anomalies or chromosomal abnormalities. Mean initial cyst diameter was 3.4 +/- 1.7 cm (+/-SD; range, 1 to 6.1). Indications used for ovarian cyst decompression included anechoic cysts with a diameter > or =4 cm, a cyst "wandering" about the abdomen on serial sonograms, or demonstrating rapid enlargement (>1 cm/wk). RESULTS: All but one cyst progressed in size during observation. One fetal ovarian cyst (diameter, 2 cm) subsequently regressed spontaneously and another (diameter, 2.1 cm) stabilized during prenatal ultrasound surveillance. One "cyst" observed with a diameter of 3.5 cm proved to be a persistent cloaca. Four fetal ovarian cysts met criteria for decompression. Because of fetal position, decompression could not be performed in one. One cyst (seen before defining criteria for decompression) with a diameter of 5 cm was observed only and underwent torsion. Two cysts (diameters, 6.1 cm and 4 cm) were decompressed in utero under local anesthesia with ultrasound guidance, of 95 mL and 35 mL, respectively. High cyst fluid progesterone (12,041 and 1,990 ng/dL, respectively) and testosterone (1,298 and 2,900 ng/dL, respectively) confirmed the etiology of the cyst as ovarian. Neither cyst recurred, and postnatal ultrasound scan confirmed resolution. There was no maternal or fetal morbidity or mortality and only the patient observed before development of criteria for decompression lost her ovary because of torsion. CONCLUSIONS: Fetal ovarian cysts tend to present as isolated unilateral lesions in normal fetuses in the third trimester. Spontaneous regression of fetal ovarian cysts may occur. Fetal ovarian cyst decompression, in select cases, may preserve ovaries at risk for torsion. PMID- 9349766 TI - Further experience with the antireflux valve to prevent ascending cholangitis in biliary atresia. AB - BACKGROUND/PURPOSE: The intussusception-type antireflux valve is a mechanism introduced by Nakajo, who reported the results of his initial 17 cases in 1990. This report summarizes our further experience with new patients, together with follow-up results of the cases previously reported by Nakajo. METHODS: In total, 46 new patients who had biliary atresia underwent portoenterostomy at the authors' two units. The authors hoped to construct the intussuscepted antireflux valve in the Roux-en-Y loop, whenever possible, at the time of hepatic portoenterostomy. RESULTS: Among the 46 patients, one case each was found to be too young or too old to have the valve constructed during the surgery. In another case, the Roux-en-Y loop became too short after repeated revisions of the portoenterostomy. In another patient, the valve was first constructed but later removed because of jejunal perforation near the valve. In the last patient, the valve was not constructed for unspecified reasons. The authors constructed the antireflux valve in 42 patients, but it was maintained in 41. In one case, the valve truly prevented reflux of the intestinal juice during an episode of intestinal obstruction. The valve was found to be incompetent in one patient 5 years after the initial surgery. CONCLUSION: The incidence of cholangitis was high in patients with postoperative cystic dilatation of the intrahepatic bile ducts, and low in those without it. PMID- 9349767 TI - 13 pairs of ribs--a predictor of long gap atresia in tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: Results of treatment of tracheoesophageal fistula (TEF) have improved over the years. However, long gap atresias continue to be a problem and require modification in the conventional operation. Preoperative diagnosis of a long gap atresia in a case of TEF is difficult, and often the defect is noted only at thoracotomy, thus necessitating multiple intraoperative changes in the position of the neonate. In the past 5 years the authors have treated 61 cases of TEF. Of these, 12 had a long gap atresia. Nine of these 12 patients had 13 pairs of ribs. None of the patients with a short gap atresia had 13 pairs of ribs. Hence, the presence of 13 pairs of ribs is a good indicator of long gap atresia. In a child who has TEF with 13 pairs of ribs, suitable modifications in operative procedure can be planned. The report also discusses the possible embryological basis of this association. PMID- 9349768 TI - Fetal adrenal transplantation: success of a laparoscopic technique in rats. AB - BACKGROUND/PURPOSE: The present study investigates a new laparoscopic technique for fetal adrenal transplantation in rats. RESULTS: The procedure was successful in 9 of 10 cases (one hole in the omentum) with no postoperative complications. On examination 4 weeks postoperatively, all but one graft showed macroscopic integrity, vascular supply, and histological maturation to normal zonal differentiation. When bilateral adrenalectomy was performed in the recipient to assess endocrine competence of the fetal adrenal grafts, survival was prolonged and Addison crisis was prevented in the animals that underwent transplantation. Levels of aldosterone dropped within the first week after adrenalectomy, but recovered steadily. Analysis of corticosterone demonstrated that levels fell to 25% of sham operated rats in the first week, but then steadily climbed to 70%. CONCLUSIONS: To the authors' knowledge this report presents the first study for laparoscopic transplantation of fetal tissues. Laparoscopic transplantation of fetal adrenal glands seemed feasible and successful in rats. The fetal adrenal transplants matured and served for a prolonged survival. PMID- 9349769 TI - Amniotic membranes as prosthetic material: experimental utilization data of a rat model. AB - BACKGROUND/PURPOSE: Prosthetic materials are applied for closing big tissue defects, the repair of traumatized organs, or hernias. Because nonabsorbable synthetic materials are rigid, possess a defined and unchangeable size, and foreign body reaction (FBR) may occur, biological materials may be an alternative. METHODS: In experimental studies in rats the authors implanted the fetal parts of the human amniotic membranes and examined the utilization and FBR induced in a standardized model. In addition amnion (AM) was combined with vicryl net (VN) for higher implant stability. Fifteen, 30 and 90 days after implantation, macroscopic appearance was examined, and light microscopy and immunohistology testing of the specimens were performed. RESULTS: Adhesions to parenchymal organs and omentum were present irrespective of the side facing the abdominal cavity. AM induced a rapid FBR, which diminished with time. Chorion (CH) and parts of the AM were resorbed within the examined period after infiltration with recipient cells and neovascularisation. The combined implant, AM, and VN showed best results because disadvantages of one material could be compensated for by the advantages of the other. CONCLUSION: The studies show that AM, in its anatomic definition, combined with VN proves to be a safe and reliable prosthetic material for the use in tissue defects. PMID- 9349770 TI - Changes in the management of pediatric blunt splenic and hepatic injuries. AB - BACKGROUND/PURPOSE: Intensive care monitoring, blood replacement, and nonoperative treatment of splenic and hepatic injuries in stable patients is the standard practice in pediatric surgery with a success rate of 90% in children's trauma centers. METHODS: During the past 5 years, 55 children under 14 years of age have been treated for laceration of spleen, liver, or both, proven by computed tomography. RESULTS: In 34 (62%), other injuries were identified, and only 21 (38%) presented with isolated injuries. In the 21 children who had isolated injuries, 18 had laceration of spleen, two had liver lacerations, and one had liver and spleen laceration. One patient who had spleen laceration required laparotomy and splenorrhaphy because of hemodynamic instability 4 hours after admission. The other 20 patients were initially closely monitored indiscriminately in the Intensive Care Unit of the pediatric surgical nursing unit. Blood transfusion was given to four children during the first 24 hours of admission despite the fact that, retrospectively, all were hemodynamically stable. There was no morbidity or mortality in all the 55 children. CONCLUSIONS: The results of this study suggest that intensive care monitoring is not mandatory in hemodynamically stable patients who have isolated liver or spleen injuries. Blood replacement should be indicated in patients who have hematocrit levels lower than 20% and signs of continuing blood loss. Because of structural characteristics of the young liver and spleen, early progressive mobilization can be indicated. PMID- 9349771 TI - A model of hypoxia-induced necrotizing enterocolitis: the role of distension. AB - BACKGROUND/PURPOSE: This study was performed to investigate additional effects of intestinal distension in the damage to the gut caused by hypoxia-reperfusion. METHODS: Five groups each consisting of ten 1-day-old Wistar albino rat pups were studied; Group 1, hypoxia-reoxygenation; Group 2, hypoxia-reoxygenation and distension; Group 3, distension and hypoxia-reoxygenation; Group 4, distension; and Group 5, control. Hypoxia was induced by placing the rat pups in a 100% carbon dioxide chamber for 5 minutes. After the hypoxia, the pups were exposed to 100% oxygen for reoxygenation for 5 minutes. The intestinal distension was carried out with a fine 21-gauge SILASTIC cannula via rectal route. The rats were killed on the third day, and all the intestine were harvested from duodenum to sigmoid colon. Malondialdehyde (MDA) levels were determined as an index of lipid peroxidation related to free radical reaction in the intestine. The histopathologic investigation was carried out by light microscopy in a blinded fashion. RESULTS: The MDA levels of Group 3 animals were significantly higher than those in Group 1, 4, and the control group (P < .05). The MDA level of Group 2 did not differ significantly from that of the Group 3 (P > .05). All MDA levels of the study groups also were significantly higher than the control group (P < .05). CONCLUSION: The results demonstrated that intestinal distension increased the damaging effects of hypoxia-reoxygenation in the gut. PMID- 9349772 TI - Laparoscopy in the management of pediatric varicoceles. AB - BACKGROUND/PURPOSE: The authors present their early experience with laparoscopic ligation of varicoceles in children. METHODS: Over a period of 30 months, 17 children underwent laparoscopic treatment of unilateral varicoceles (age range, 7 to 16 years). Nine underwent ligation of veins alone and eight (including four children who had recurrent varicoceles) underwent ligation of testicular veins and artery. In three patients, vas-associated veins were ligated in addition to the main testicular vessels. RESULTS: The average operation time was 30 minuts (range, 14 to 75) and hospital stay, 11 hours (range, 4 to 22). There were no technical failures. Minor scrotal emphysema developed in one child, and a painful, tender swelling above the epididymis developed in another. At an average follow-up of 24 months (range, 6 to 30), all patients were asymptomatic and had marked reduction in the size of the varicoceles. CONCLUSION: Further experience and long-term follow-up are clearly required before the efficacy of this procedure is established. PMID- 9349773 TI - Treatment of loculated pleural effusion with intrapleural urokinase in children. AB - BACKGROUND/PURPOSE: The use of fibrinolytic agents such as urokinase and streptokinase has been reported in cases of empyema in adults. In pediatric patients the experience is, however, very limited. METHODS: A series of seven consecutive children who had loculated pleural effusion that did not respond to drainage and antibiotics is reported. RESULTS: In all cases, the effusion was found to be multiloculated. Urokinase (UK) instillation through the already existing chest tube was started. A dose of 100,000 U of UK diluted in 100 mL of normal saline was instilled through the chest tube, which was clamped for 12 hours and then was left open for another 12 hours. In six of seven children, the treatment was terminated after complete or almost complete resolution was attained. This was achieved within 5 treatment days (mean, 3.3). There was one failure which was attributed to relatively late initiation of treatment. No complications were observed. CONCLUSIONS: The authors conclude that intrapleural administration of UK is a safe and efficient method of treatment in cases of loculated pleural effusions in children. UK instillation to the intrapleural space should be considered early before initiating surgical intervention. Starting intrapleural UK treatment should not be delayed. PMID- 9349774 TI - Fetus-in-fetu with malignant recurrence. AB - Fetus-in-fetu is an unusual condition in which a vertebrate fetus is enclosed within the abdomen of another fetus. These occurrences are usually benign. This report describes an instance of malignant recurrence after resection of a fetus in-fetu. PMID- 9349775 TI - Pseudoexstrophy of the bladder: case report and literature review. AB - The authors present a case of a boy newborn who had bladder pseudoexstrophy and multiple congenital anomalies. In addition to this unusual variant of the exstrophy-epispadias complex, the patient was found to have a posterior cleft palate, an omphalocele, and an imperforate anus. PMID- 9349776 TI - Syndactyly repair performed simultaneously with circumcision: use of foreskin as a skin-graft donor site. AB - A boy who had simple syndactyly involving the third web space of the left hand presented for elective syndactyly repair. Circumcision had been delayed because of neonatal medical problems. Elective syndactyly repair and circumcision were performed in one operation at age 9 months. Penile foreskin was used as a full thickness skin graft for the syndactyly repair. The foreskin provided a functional syndactyly repair with good aesthetic characteristics. This obviated the need for two separate operations and for an additional skin graft donor site. To our knowledge, this is the first reported case in which foreskin was used for the repair of syndactyly. In boys with syndactyly, the authors advocate that parents be informed of this reconstructive option. Should the parents consider it to be suitable, then elective circumcision should be delayed until the time of syndactyly repair so that foreskin may be used for the syndactyly repair. PMID- 9349778 TI - Primary reconstruction of esophageal atresia with distal tracheoesophageal fistula in a 740-g infant. AB - A premature 740-g infant who had esophageal atresia and tracheoesophageal fistula was treated with a primary anastomosis. The postoperative recovery was excellent, but the need for close cooperation with neonatal intensivists is essential for survival. PMID- 9349777 TI - Congenital esophageal stenosis owing to tracheobronchial remnants: a case report. AB - A 3.5-year-old girl who had congenital esophageal stenosis caused by tracheobronchial remnants without esophageal atresia or tracheoesophageal fistula is presented. The diagnostic difficulties are discussed. PMID- 9349779 TI - Successful management of ruptured choledochal cyst by primary cyst excision and biliary reconstruction. AB - One to two percent of patients who have choledochal cysts present with cyst rupture and bile peritonitis. Reported cases have been managed with external drainage of the cyst followed by a second procedure to excise the cyst and reconstruct the biliary tract. The authors report two cases of ruptured choledochal cyst treated with primary cyst excision and biliary drainage. The satisfactory outcome of these patients suggests that this is the preferred management. PMID- 9349780 TI - Unusual presentation of heteropagus attached to the thorax. AB - The endoparasitic twin is the most common form of asymmetric fetal duplication (heteropagus). A 2-month-old girl who had a parasitic right lower limb with axial skeleton, vertebral column, uterus, fallopian tube, ovary, and bladder implanted in the sternum region is described as another example of exoparasitic twin, the uncommon form of heteropagus. Unusually, dextrocardia was found in the autosite. This report emphasizes the even progression between the endoparasitic and exoparasitic forms of heteropagus. PMID- 9349781 TI - Conservative management of mesenchymal hamartoma of the liver. AB - The natural history of mesenchymal hamartoma of the liver is poorly understood. This case demonstrates the course of a biopsy-proven mesenchymal hamartoma using sequential computed tomography (CT) examinations. These CT scans show initial expansion of the lesion with subsequent involution. The spontaneous resolution in this patient suggests the possibility of conservative management of asymptomatic mesenchymal hamartomas. The case is presented, and the literature on mesenchymal hamartoma is reviewed. PMID- 9349782 TI - An extremely rare variant of congenital jejunoileo-colic atresia. AB - This report presents an unusual case of a congenital long-segment jejunoileo colic obstruction without mesenteric or intestinal interruption. Histologically, the intestinal lumen was completely or partially obstructed by fibrous or granulation tissue, and the mucosa had disappeared. PMID- 9349783 TI - Ileal atresia with total colonic aganglionosis. AB - This is the 17th report of the case of an infant who had ileal atresia associated with Hirschsprung's disease, and the second with ganglion cells distal to the atresia. Experience suggests that Hirschsprung's disease should be suspected in all forms of bowel atresia. PMID- 9349784 TI - Failed Nissen fundoplication in two patients who had persistent vomiting and eosinophilic esophagitis. AB - The following report describes two patients who had chronic symptoms of gastroesophageal reflux and persistent histological esophagitis, despite aggressive medical antireflux therapy, who continued to have esophagitis and remained symptomatic post antireflux surgery (Nissen fundoplication). Both patients demonstrated a severe eosinophilic esophagitis with normal gastric and duodenal histology before and after surgery. Postoperatively, each received the diagnosis of allergic enteritis and both responded clinically and histologically to oral corticosteroids and an elemental diet. PMID- 9349786 TI - Furuncular cuterebrid myiasis. AB - Myiasis is the infestation of skin bythe larvae of flies. In North America cases are caused by the botfly (Cuterebra) and occur most commonly in children. The usual presentation is a subcutaneous abscess, and for this reason these patients may be referred to surgeons. Knowledge of this entity can avoid delays in diagnosis, unnecessary incision and drainage procedures, and unnecessary courses of antibiotics. PMID- 9349785 TI - Detection of tumor thrombus in children using color Doppler ultrasonography. AB - Color Doppler ultrasonography (CDUS), which provides information regarding both the tissue and the blood flow simultaneously in real time, is a relatively new and noninvasive technique for the evaluation of organ perfusion. The authors investigated the usefulness of CDUS in the detection of tumor thrombus in 34 children who had malignant tumor (21 neuroblastomas, 2 Wilms' tumors, 5 hepatoblastomas, 2 rhabdomyosarcomas, and 4 others) who underwent CDUS between April 1992 and March 1995 in our department. The CDUS showed tumor thrombus in four patients (11.8%). Three thrombi (in one patient each with neuroblastoma, Wilms' tumor, and hepatoblastoma) were situated in the inferior vena cava, and one (in a patient who had hepatoblastoma) in the portal vein. Each tumor thrombus was monitored and followed up using CDUS every month during the entire clinical course; two of the four thrombi remained despite intensive treatments. The authors have found CDUS to afford rapid and noninvasive detection of tumor thrombus, and it proved useful not only in the detection of tumor thrombus but also in its follow-up in these children who had malignant tumor. PMID- 9349787 TI - A case of ileosigmoid knotting in a child. PMID- 9349788 TI - Surgical emergency embolectomy for the treatment of fulminant pulmonary embolism in a preterm infant. AB - A massive pulmonary embolism, demonstrated by echocardiography developed in a 3 week-old preterm infant. An etiologic explanation could not be obtained from either history or clinical and laboratory findings. Pulmonary embolectomy was performed as an emergency procedure because of severe hemodynamic impairment despite intensive medical therapy. In children who have massive pulmonary embolism who remain in a compromised hemodynamic state despite intensive medical therapy, pulmonary embolectomy may be considered the alternative emergency treatment. PMID- 9349789 TI - Idiopathic iliac arterial aneurysms in a child. AB - Arterial aneurysms are very rare in children, especially those who have no history of cardiac or vascular malformation, connective tissue disorder, or trauma. We describe a 3-year-old boy who had multiple arterial aneurysms of the left external iliac artery with a maximal diameter of 67 mm, with no history of these disorders. The iliac artery distal to the aneurysm and superficial femoral artery were occluded causing the growth disturbances of his left leg. He underwent graft replacement of the left iliac artery using a ringed Gore-Tex graft (6 mm in diameter). Postoperative angiogram showed a patent graft without any residual aneurysm in the left iliac artery. His ankleibrachial index improve from 0.2 to 0.6 after surgery. PMID- 9349790 TI - Early presentation of choledochal cyst transiently obstructed by an inspissated bile plug. PMID- 9349791 TI - Cure of hypoglycemic hyperinsulinism by enucleation of a focal islet cell adenomatous hyperplasia. AB - During pancreatectomy for refractory neonatal hyperinsulinemic hypoglycemia, a well-delineated focal adenomatous hyperplasia was enucleated. Intraoperative glucose levels returned to normal and pancreatectomy was averted. Seven months later the child is euglycemic. This experience suggests that during surgery for neonatal refractory hypoglycemia, a focal lesion should be sought, and if found, enucleated, and blood glucose monitored. If the glucose rises to euglycemic levels or above, the child should be monitored clinically. If sustained elevation is not maintained, a search for an additional focal lesion or pancreatectomy should be performed. Saving the pancreas may prevent future development of diabetes mellitus. PMID- 9349792 TI - Light in dark places. PMID- 9349793 TI - Nobel prizes honour biologists' work on protein energy converters. PMID- 9349794 TI - Genome research strategy splits Japanese scientists. PMID- 9349796 TI - Diversity project 'does not merit federal funding'. PMID- 9349795 TI - Genome study maps chemical sensitivity. PMID- 9349797 TI - 'Bioethics needs better input from public'. PMID- 9349798 TI - Nobel backing for alternative health journal. PMID- 9349799 TI - All sorts of authorship. PMID- 9349800 TI - Value for money in US laboratories. PMID- 9349801 TI - Some like it hot: spicing up ion channels. PMID- 9349802 TI - New age crystals. PMID- 9349803 TI - Disomy and disease resolved? PMID- 9349804 TI - Neurotrophins moving forward. PMID- 9349805 TI - Synapses. Plastic plasticity. PMID- 9349806 TI - Hans Jurgen Eysenck (1916-97) PMID- 9349808 TI - Sex on the brain. PMID- 9349807 TI - Prion research: the next frontiers. PMID- 9349809 TI - Antibiotic resistance spread in food. PMID- 9349810 TI - Targeted disruption in Arabidopsis. PMID- 9349811 TI - Extraterrestrial handedness. PMID- 9349812 TI - Transactivation of Igf2 in a mouse model of Beckwith-Wiedemann syndrome. AB - The gene IGF2, which encodes a fetal insulin-like growth factor, is imprinted, so only one of two parental copies of the gene is expressed. The altered expression of IGF2 has been implicated in Beckwith-Wiedemann syndrome, a human fetal overgrowth syndrome, which is characterized by overgrowth of several organs and an increased risk of developing childhood tumours. We have introduced Igf2 transgenes into the mouse genome by using embryonic stem cells, which leads to transactivation of the endogenous Igf2 gene. The consequent overexpression of Igf2 results in most of the symptoms of Beckwith-Wiedemann syndrome, including prenatal overgrowth, polyhydramnios, fetal and neonatal lethality, disproportionate organ overgrowth including tongue enlargement, and skeletal abnormalities. These phenotypes establish Igf2 overexpression as a key determinant of Beckwith-Wiedemann syndrome. PMID- 9349813 TI - The capsaicin receptor: a heat-activated ion channel in the pain pathway. AB - Capsaicin, the main pungent ingredient in 'hot' chilli peppers, elicits a sensation of burning pain by selectively activating sensory neurons that convey information about noxious stimuli to the central nervous system. We have used an expression cloning strategy based on calcium influx to isolate a functional cDNA encoding a capsaicin receptor from sensory neurons. This receptor is a non selective cation channel that is structurally related to members of the TRP family of ion channels. The cloned capsaicin receptor is also activated by increases in temperature in the noxious range, suggesting that it functions as a transducer of painful thermal stimuli in vivo. PMID- 9349815 TI - Spatial invariance of visual receptive fields in parietal cortex neurons. AB - Spatial information is conveyed to the primary visual cortex in retinal coordinates. Movement trajectory programming, however, requires a transformation from this sensory frame of reference into a frame appropriate for the selected part of the body, such as the eye, head or arms. To achieve this transformation, visual information must be combined with information from other sources: for instance, the location of an object of interest can be defined with respect to the observer's head if the position of the eyes in the orbit is known and is added to the object's retinal coordinates. Here we show that in a subdivision of the monkey parietal lobe, the ventral intraparietal area (VIP), the activity of visual neurons is modulated by eye-position signals, as in many other areas of the cortical visual system. We find that individual receptive fields of a population of VIP neurons are organized along a continuum, from eye to head coordinates. In the latter case, neurons encode the azimuth and/or elevation of a visual stimulus, independently of the direction in which the eyes are looking, thus representing spatial locations explicitly in at least a head-centred frame of reference. PMID- 9349814 TI - Polymerized colloidal crystal hydrogel films as intelligent chemical sensing materials. AB - Chemical sensors respond to the presence of a specific analyte in a variety of ways. One of the most convenient is a change in optical properties, and in particular a visually perceptible colour change. Here we report the preparation of a material that changes colour in response to a chemical signal by means of a change in diffraction (rather than absorption) properties. Our material is a crystalline colloidal array of polymer spheres (roughly 100 nm diameter) polymerized within a hydrogel that swells and shrinks reversibly in the presence of certain analytes (here metal ions and glucose). The crystalline colloidal array diffracts light at (visible) wavelengths determined by the lattice spacing, which gives rise to an intense colour. The hydrogel contains either a molecular recognition group that binds the analyte selectively (crown ethers for metal ions), or a molecular-recognition agent that reacts with the analyte selectively. These recognition events cause the gel to swell owing to an increased osmotic pressure, which increases the mean separation between the colloidal spheres and so shifts the Bragg peak of the diffracted light to longer wavelengths. We anticipate that this strategy can be used to prepare 'intelligent' materials responsive to a wide range of analytes, including viruses. PMID- 9349816 TI - False perception of motion in a patient who cannot compensate for eye movements. AB - We are usually unaware of the motion of an image across our retina that results from our own movement. For instance, during slow-tracking eye movements we do not mistake the shift of the image projected onto the retina for motion of the world around us, but instead perceive a stable world. Following early suggestions by von Helmholtz, it is commonly believed that this spatial stability is achieved by subtracting the retinal motion signal from an internal reference signal, such as a copy of the movement command (efference copy). Object motion is perceived only if the two differ. Although this concept is widely accepted, its anatomical underpinning remains unknown. Here we describe the case of a patient with bilateral extrastriate cortex lesions, suffering from false perception of motion due to an inability to take eye movements into account when faced with self induced retinal image slip. This is indicated by the fact that during smooth pursuit eye movements, he perceives motion of the stationary world at a velocity that corresponds to the velocity of his eye movement; that is, he perceives the raw retinal image slip uncorrected for his own eye movements. We suspect that this deficiency reflects damage of a distinct parieto-occipital region that disentangles self-induced and externally induced visual motion by comparing retinal signals with a reference signal encoding eye movements and possibly ego motion in general. PMID- 9349817 TI - Ligand-induced changes in integrin expression regulate neuronal adhesion and neurite outgrowth. AB - Receptors of the integrin family are expressed by every cell type and are the primary means by which cells interact with the extracellular matrix. The control of integrin expression affects a wide range of developmental and cellular processes, including the regulation of gene expression, cell adhesion, cell morphogenesis and cell migration. Here we show that the concentration of substratum-bound ligand (laminin) post-translationally regulates the amount of receptor (alpha6beta1, integrin) expressed on the surface of sensory neurons. When ligand availability is low, surface amounts of receptor increase, whereas integrin RNA and total integrin protein decrease. Ligand concentration determines surface levels of integrin by altering the rate at which receptor is removed from the cell surface. Furthermore, increased expression of integrin at the cell surface is associated with increased neuronal cell adhesion and neurite outgrowth. These results indicate that integrin regulation maintains neuronal growth-cone motility over a broad range of ligand concentrations, allowing axons to invade different tissues during development and regeneration. PMID- 9349818 TI - Anterograde transport of brain-derived neurotrophic factor and its role in the brain. AB - The role of neurotrophins as target-derived proteins that promote neuron survival following their retrograde transport from the terminals to the cell bodies of neurons has been firmly established in the developing peripheral nervous system. However, neurotrophins appear to have more diverse functions, particularly in the adult central nervous system. Brain-derived neurotrophic factor (BDNF), for example, produces a variety of neuromodulatory effects in the brain that are more consistent with local actions than with long-distance retrograde signalling. Here we show that BDNF is widely distributed in nerve terminals, even in brain areas such as the striatum that lack BDNF messenger RNA, and that inhibition of axonal transport or deafferentation depletes BDNF. The number of striatal neurons that contain the calcium-binding protein parvalbumin was decreased in BDNF+/- and BDNF /- mice in direct proportion to the loss of BDNF protein, which is consistent with anterogradely supplied BDNF having a functional role in development or maintenance. Thus the anterograde transport of BDNF from neuron cell bodies to their terminals may be important for the trafficking of BDNF in the brain. PMID- 9349819 TI - Metaplasticity at identified inhibitory synapses in Aplysia. AB - Synaptic plasticity is an important feature of neural networks involved in the encoding of information. In the analysis of long-term potentiation and long-term depression, several examples have emerged in which this plasticity is itself modulated. This higher-order form of plasticity has been referred to as 'metaplasticity', a modification of synapses reflected as a change in the ability to induce or maintain plasticity. These observations raise the question of the possible advantage of regulating the intrinsic plastic properties of a synapse. The neural circuit mediating the siphon withdrawal reflex in Aplysia provides a useful network in which to examine this question directly. Inhibitory synapses in this circuit (from L30 neurons) exhibit a variety of forms of activity-dependent short-term synaptic enhancement which contribute to dynamic gain control in the siphon withdrawal reflex. Here we report that tail shock, an extrinsic modulatory input of known behavioural relevance, induces differential metaplasticity at this synapse, attenuating its ability to exhibit short-term synaptic enhancement after presynaptic activation (augmentation and post-tetanic potentiation), while leaving intact its capacity for enhancement during activation. This attenuation of inhibition at the synaptic level seems to mediate comparable attenuation of inhibitory modulation at both network and behavioural levels. PMID- 9349820 TI - Absence of excitotoxicity-induced apoptosis in the hippocampus of mice lacking the Jnk3 gene. AB - Excitatory amino acids induce both acute membrane depolarization and latent cellular toxicity, which often leads to apoptosis in many neurological disorders. Recent studies indicate that glutamate toxicity may involve the c-Jun amino terminal kinase (JNK) group of mitogen-activated protein (MAP) kinases. One member of the JNK family, Jnk3, may be required for stress-induced neuronal apoptosis, as it is selectively expressed in the nervous system. Here we report that disruption of the gene encoding Jnk3 in mice caused the mice to be resistant to the excitotoxic glutamate-receptor agonist kainic acid: they showed a reduction in seizure activity and hippocampal neuron apoptosis was prevented. Although application of kainic acid imposed the same level of noxious stress, the phosphorylation of c-Jun and the transcriptional activity of the AP-1 transcription factor complex were markedly reduced in the mutant mice. These data indicate that the observed neuroprotection is due to the extinction of a Jnk3 mediated signalling pathway, which is an important component in the pathogenesis of glutamate neurotoxicity. PMID- 9349822 TI - Activation of the transcription factor Gli1 and the Sonic hedgehog signalling pathway in skin tumours. AB - Sporadic basal cell carcinoma (BCC) is the most common type of malignant cancer in fair-skinned adults. Familial BCCs and a fraction of sporadic BCCs have lost the function of Patched (Ptc), a Sonic hedgehog (Shh) receptor that acts negatively on this signalling pathway. Overexpression of Shh can induce BCCs in mice. Here we show that ectopic expression of the zinc-finger transcription factor Gli1 in the embryonic frog epidermis results in the development of tumours that express endogenous Gli1. We also show that Shh and the Gli genes are normally expressed in hair follicles, and that human sporadic BCCs consistently express Gli1 but not Shh or Gli3. Because Gli1, but not Gli3, acts as a target and mediator of Shh signalling, our results suggest that expression of Gli1 in basal cells induces BCC formation. Moreover, loss of Ptc or overexpression of Shh cannot be the sole causes of Gli1 induction and sporadic BCC formation, as they do not occur consistently. Thus any mutations leading to the expression of Gli1 in basal cells are predicted to induce BCC formation. PMID- 9349821 TI - Identification and characterization of the vesicular GABA transporter. AB - Synaptic transmission involves the regulated exocytosis of vesicles filled with neurotransmitter. Classical transmitters are synthesized in the cytoplasm, and so must be transported into synaptic vesicles. Although the vesicular transporters for monoamines and acetylcholine have been identified, the proteins responsible for packaging the primary inhibitory and excitatory transmitters, gamma aminobutyric acid (GABA) and glutamate remain unknown. Studies in the nematode Caenorhabditis elegans have implicated the gene unc-47 in the release of GABA. Here we show that the sequence of unc-47 predicts a protein with ten transmembrane domains, that the gene is expressed by GABA neurons, and that the protein colocalizes with synaptic vesicles. Further, a rat homologue of unc-47 is expressed by central GABA neurons and confers vesicular GABA transport in transfected cells with kinetics and substrate specificity similar to those previously reported for synaptic vesicles from the brain. Comparison of this vesicular GABA transporter (VGAT) with a vesicular transporter for monoamines shows that there are differences in the bioenergetic dependence of transport, and these presumably account for the differences in structure. Thus VGAT is the first of a new family of neurotransmitter transporters. PMID- 9349823 TI - A SNARE involved in protein transport through the Golgi apparatus. AB - In eukaryotic cells, the Golgi apparatus receives newly synthesized proteins from the endoplasmic reticulum (ER) and delivers them after covalent modification to their destination in the cell. These proteins move from the inside (cis) face to the plasma-membrane side (trans) of the Golgi, through a stack of cisternae, towards the trans-Golgi network (TGN), but very little is known about how proteins are moved through the Golgi compartments. In a model known as the maturation model, no special transport process was considered necessary, with protein movement along the Golgi being achieved by maturation of the cisternae. Alternatively, proteins could be transported by vesicles or membrane tubules. Although little is known about membrane-tubule-mediated transport, the molecular mechanism for vesicle-mediated transport is quite well understood, occurring through docking of SNAREs on the vesicle with those on the target membrane. We have now identified a protein of relative molecular mass 27K which is associated with the Golgi apparatus. The cytoplasmic domain of this protein or antibodies raised against it quantitatively inhibit transport in vitro from the ER to the trans-Golgi/TGN, acting at a stage between the cis/medial- and the trans Golgi/TGN. This protein, which behaves like a SNARE and has been named GS27 (for Golgi SNARE of 27K), is identical to membrin, a protein implicated earlier in ER to-Golgi transport. Our results suggest that protein movement from medial- to the trans-Golgi/TGN depends on SNARE-mediated vesicular transport. PMID- 9349824 TI - Crystallographic structure of the T domain-DNA complex of the Brachyury transcription factor. AB - The mouse Brachyury (T) gene is the prototype of a growing family of so-called T box genes which encode transcriptional regulators and have been identified in a variety of invertebrates and vertebrates, including humans. Mutations in Brachyury and other T-box genes result in drastic embryonic phenotypes, indicating that T-box gene products are essential in tissue specification, morphogenesis and organogenesis. The T-box encodes a DNA-binding domain of about 180 amino-acid residues, the T domain. Here we report the X-ray structure of the T domain from Xenopus laevis in complex with a 24-nucleotide palindromic DNA duplex. We show that the protein is bound as a dimer, interacting with the major and the minor grooves of the DNA. A new type of specific DNA contact is seen, in which a carboxy-terminal helix is deeply embedded into an enlarged minor groove without bending the DNA. Hydrophobic interactions and an unusual main-chain carbonyl contact to a guanine account for sequence-specific recognition in the minor groove by this helix. Thus the structure of this T domain complex with DNA reveals a new way in which a protein can recognize DNA. PMID- 9349825 TI - The semi-individual study in air pollution epidemiology: a valid design as compared to ecologic studies. AB - The assessment of long-term effects of air pollution in humans relies on epidemiologic studies. A widely used design consists of cross-sectional or cohort studies in which ecologic assignment of exposure, based on a fixed-site ambient monitor, is employed. Although health outcome and usually a large number of covariates are measured in individuals, these studies are often called ecological. We will introduce the term semi-individual design for these studies. We review the major properties and limitations with regard to causal inference of truly ecologic studies, in which outcome, exposure, and covariates are available on an aggregate level only. Misclassification problems and issues related to confounding and model specification in truly ecologic studies limit etiologic inference to individuals. In contrast, the semi-individual study shares its methodological and inferential properties with typical individual-level study designs. The major caveat relates to the case where too few study areas, e.g., two or three, are used, which render control of aggregate level confounding impossible. The issue of exposure misclassification is of general concern in epidemiology and not an exclusive problem of the semi-individual design. In a multicenter setting, the semi-individual study is a valuable tool to approach long-term effects of air pollution. Knowledge about the error structure of the ecologically assigned exposure allows consideration of the impact of ecologically assigned exposure on effect estimation. Semi-individual studies, i.e., individual level air pollution studies with ecologic exposure assignment, more readily permit valid inference to individuals and should not be labeled as ecologic studies. PMID- 9349826 TI - Artifacts associated with the measurement of oxidized DNA bases. AB - In this paper we review recent aspects of the measurement of oxidized DNA bases, currently a matter of debate. There has long been an interest in the determination of the level of oxidized bases in cellular DNA under both normal and oxidative stress conditions. In this respect, the situation is confusing because variations that may be as large as two orders of magnitude have been reported for the yield of the formation of 8-oxo-7,8-dihydroguanine (8-oxoGua) in similar DNA samples. However, recent findings clearly show that application of several assays like gas chromatography-mass spectrometry (GC-MS) and -32P- postlabeling may lead to a significant overestimation of the level of oxidized bases in cellular DNA. In particular, the silylation step, which is required to make the samples volatile for the GC-MS analysis, has been shown to induce oxidation of normal bases at the level of about one oxidized base per 10(4) normal bases. This has been found to be a general process that applies in particular to 8-oxoGua, 8-oxo-7, 8-dihydroadenine,5-hydroxycytosine, 5 (hydroxymethyl)uracil, and 5-formyluracil. Interestingly, prepurification of the oxidized bases from DNA hydrolysate prior to the derivatization reaction prevents artefactual oxidation. Under these conditions, the level of oxidized bases measured by GC-MS is similar to that obtained by HPLC associated with electrochemical detection (HPLC-EC). It should be added that the level of 8-oxo 7,8-dihydro-2;-deoxyguanosine in control cellular DNA has been found to be about fivefold lower than in earlier HPLC-EC measurements by using appropriate conditions of extraction and enzymatic digestion of DNA. Similar conclusions were reached by measuring formamidopyrimidine-DNA glycosylase sensitive sites as revealed by the single cell gel electrophoresis (comet) assay. PMID- 9349827 TI - Inorganic arsenic: a need and an opportunity to improve risk assessment. AB - This paper presents views on the current status of (inorganic) arsenic risk assessment in the United States and recommends research needed to set standards for drinking water. The opinions are those of the Arsenic Task Force of the Society for Environmental Geochemistry and Health, which has met periodically since 1991 to study issues related to arsenic risk assessment and has held workshops and international conferences on arsenic. The topic of this paper is made timely by current scientific interest in exposure to and adverse health effects of arsenic in the United States and passage of the Safe Drinking Water Act Amendment of 1996, which has provisions for a research program on arsenic and a schedule mandating the EPA to revise the maximum contaminant level of arsenic in drinking water by the year 2001. Our central premise and recommendations are straightforward: the risk of adverse health effects associated with arsenic in drinking water is unknown for low arsenic concentrations found in the United States, such as at the current interim maximum contaminant level of 50 microg/l and below. Arsenic-related research should be directed at answering that question. New epidemiological studies are needed to provide data for reliable dose-response assessments of arsenic and for skin cancer, bladder cancer, or other endpoints to be used by the EPA for regulation. Further toxicological research, along with the observational data from epidemiology, is needed to determine if the dose-response relationship at low levels is more consistent with the current assumption of low-dose linearity or the existence of a practical threshold. Other recommendations include adding foodborne arsenic to the calculation of total arsenic intake, calculation of total arsenic intake, and encouraging cooperative research within the United States and between the United States and affected countries. PMID- 9349829 TI - Environmental Health Information Service. PMID- 9349828 TI - Pesticides and childhood cancers. AB - To evaluate the possible association between pesticides and the risk of childhood cancers, epidemiologic studies published between 1970 and 1996 were critically reviewed. Thirty-one studies investigated whether occupational or residential exposure to pesticides by either parents or children was related to increased risk of childhood cancer. In general, the reported relative risk estimates were modest. Risk estimates appeared to be stronger when pesticide exposure was measured in more detail. Frequent occupational exposure to pesticides or home pesticide use was more strongly associated with both childhood leukemia and brain cancer than either professional exterminations or the use of garden pesticides. Occupational pesticide exposure was also associated with increased risk of Wilms' tumor, Ewing's sarcoma, and germ cell tumors. Residence on a farm, a proxy for pesticide exposure, was associated with increased risk of a number of childhood cancers. Although increased risk of some childhood cancers in association with pesticide exposure is suggested by multiple studies, methodological limitations common to many studies restrict conclusions; these include indirect exposure classification, small sample size, and potential biases in control selection. Opportunities for methodologic improvement in future studies of pesticides and childhood cancers are described. PMID- 9349830 TI - Is it worth a dam? AB - Once a sign of modernization and growth, dams are often seen today as symbols of environmental and social devastation. Over 800,000 dams have been built worldwide to provide drinking water, flood control, hydropower, irrigation, navigation, and water storage. Dams do indeed provide these things,but at the cost of several adverse, unexpected effects: disruption of ecosystems, decline of fish stocks, forced human and animal resettlements, and diseases such as malaria, which are borne by vectors that thrive in quiet waters. PMID- 9349831 TI - Behavioral changes following participation in a home health promotional program in King County, Washington. AB - This study examined behavioral changes in households after participation in a home environmental assessment. Home assessment visits by a trained coach, which involved a walk-through in the home with the home residents, were conducted in 36 homes. The walk-through included a list of recommended behavioral changes that the residents could make to reduce their exposures to home pollutants in areas such as dust control, moisture problems, indoor air, hazardous household products, and hobbies. Recruited households were surveyed 3 months after the home assessment to evaluate their implementation of the recommendations. Following the home visits, 31 of 36 households reported making at least one behavioral change, and 41% of the recommendations made by the volunteer coaches were implemented. In conclusion, this study found that the majority of the households who participated in the home assessment reported implementing at least one recommendation. This home health promotional method was effective in influencing behavioral changes. PMID- 9349833 TI - Cadmium burden of men and women who report regular consumption of confectionery sunflower kernels containing a natural abundance of cadmium. AB - Because of inherent genetic and physiological characteristics, the natural concentration of cadmium in the kernels of sunflowers grown in uncontaminated soils of the northern Great Plains region of the United States is higher than in most other grains. We tested the hypothesis that a habitual consumption of sunflower kernels will increase the body burden and health effects of cadmium in humans. Sixty-six men and women who reported consuming various amounts of sunflower kernels were recruited and divided by sex and kernel consumption: those who consumed less than or equal to 1 ounce(oz)/week and those who consumed more than 1 oz/week. Cadmium intake was assessed by calculation from 7-day food diaries, cadmium burden by whole blood cadmium, red blood cell (RBC) cadmium and urine cadmium concentrations, and health effects by urinary excretion of N-acetyl beta-D-glucosaminidase (NAG) activity and beta2-microglobulin (beta2MG). The results showed that high intakes of sunflower kernels (>1 oz/day) significantly increased the intake of cadmium (p<0.004). However, the amount of cadmium in whole blood or RBCs was not affected by cadmium intake. Urinary excretion of cadmium also was not affected by cadmium intake. Urine NAG activity and the amount of urinary beta2MG were significantly elevated in the urine of high sunflower kernel consumers when the values were expressed on a urine volume basis (p<0.03), but not when expressed on a creatinine basis (p>0.05). Because normal ranges for the excretion of these protein markers have not been established, it was not possible to determine if these elevated values were meaningful. However, given the knowledge that habitual consumption of sunflower kernels with natural cadmium concentrations higher than most other food products will increase the average intake of dietary cadmium, the potential exists for an increased body burden of cadmium. Controlled feeding studies in humans should be pursued in order to determine if the body burden does indeed increase and, if so, is it a cause for concern. PMID- 9349832 TI - Children with moderately elevated blood lead levels: a role for other diagnostic tests? AB - In this study we examined potential limitations of relying exclusively on blood lead (BPb) levels to evaluate children with moderately elevated BPb levels (1.21 2.12 micromol/l, or 25-44 microg/dl). We tested the following hypotheses: 1) such children without elevated erythrocyte protoporphyrin (EP) levels (>=0.62 micromol/l or >/= 35 microg/dl) are unlikely to respond to a chelating agent with a brisk urinary Pb diuresis; 2) those with elevated EP levels, but low hematologic indices consistent with iron deficiency, are also unlikely to respond to a chelating agent with a robust urinary Pb diuresis; and 3) those with elevated EP levels and iron sufficiency are more likely to respond to a chelating agent. To test these hypotheses, we performed retrospective analyses of the relationships between EP concentrations, hematologic indices, and urinary Pb excretion ratios (uPbr) in moderately Pb-poisoned children undergoing the CaNa2EDTA lead mobilization test (Pb-MT). Data from 122 children were available. Urinary Pb excretion was limited in children with an EP <0.62 micromol/l (<35 microg/dl); only 5% (1/21) of Pb-MTs were positive (uPbr >=0.6). In children with an EP >=0.62 micromol/l, low hematologic indices, such as a mean corpuscular hemoglobin (MCH) <23 pg, were associated with relatively little Pb excretion (0/14 positive Pb-MTs). In contrast, 32% (28/87) of Pb-MTs were positive in children with an EP >/= 0.62 micromol/l and iron sufficiency (p<0.01 by chi square comparison between groups with EP >/= 0.62 micromol/l and either MCH <23 pg or MCH >/= 23 pg). We conclude that only a minority of moderately Pb-poisoned children will demonstrate enhanced urinary Pb excretion in response to chelation therapy. Some of the predicted nonresponders can be readily identified by adding the EP and complete blood count to the panel of tests performed. PMID- 9349835 TI - Reproductive toxins and alligator abnormalities at Lake Apopka, Florida. AB - The alligator population at Lake Apopka in central Florida declined dramatically between 1980 and 1987. Endocrine-disrupting chemicals and specifically DDT metabolites have been implicated in the alligators' reproductive failure. The DDT metabolite hypothesis is based largely on the observation of elevated concentrations of p,p-DDE and p,p-DDD in alligator eggs obtained from Lake Apopka in 1984 and 1985. In the following commentary, we draw attention to two nematocides that are established reproductive toxins in humans, dibromochloropropane (DBCP) and ethylene dibromide (EDB), which could also have played a role in the reproductive failure observed in alligators from Lake Apopka in the early 1980s. PMID- 9349836 TI - Global health: what's in it for us? AB - A new report, America's Vital Interest in Global Health, released in June by the Institute of Medicine's (IOM) Board on International Health, calls for increased U.S. foreign health care spending to fund research and education about diseases of the developing world, a global surveillance system to spot environmental changes and emerging disease conditions, public and private sector partnerships to distribute vaccines and drugs overseas, and a new government body to help coordinate these efforts. The report argues that, in an increasingly global society, the United States can't afford to ignore its neighbors' problems, for economic as well as social reasons. PMID- 9349834 TI - Hair analysis does not support hypothesized arsenic and chromium exposure from drinking water in Woburn, Massachusetts. AB - We hypothesized that residents of Woburn, Massachusetts, had been exposed to as much as 70 microg/l of arsenic (As) and 240 microg/l of chromium (Cr) in drinking water from municipal supply wells G and H. To test this hypothesis, we measured the concentrations of As and Cr in 82 hair samples donated by 56 Woburn residents. Thirty-six samples were cut between 1964 and 1979, the period during which wells G and H were in operation. The remainder were cut either before 1964 (1938-1963; n = 26) or after 1979 (1982-1994; n = 20). Washed hair samples were analyzed by instrumental neutron activation. Exposure to the well water--measured as access--was estimated using well pumping records and a model of the Woburn water distribution system. Our results show that access to wells G and H water was not significantly correlated (95% confidence interval) with As and Cr concentrations measured in the hair of Woburn residents, but As concentrations have declined significantly over the last half century. Linear regression of As concentrations (micrograms per gram) upon year of hair cut and access to wells G and H water yielded a standard coefficient for year of -0. 0074 +/- 0.0017 (standard error; p = 2.5 -multiple- 10(-5)) and -0.12 +/- 0.10 (p = 0.22) for access. The r2 value for the model was 0.19. The geometric mean concentrations (geometric standard deviation) of As and Cr in the hair of residents who had access (i.e., relative access estimate >0) to wells G and H water (n = 27) were 0.14 (2.6) and 2.29 (1.8) microg/g, respectively; the geometric mean concentrations of As and Cr in all of the hair samples from residents who did not have access (1938-1994; n = 55) were 0.13 (3.0) and 2.19 (2.0) microg/g, respectively. PMID- 9349837 TI - Thyroid hormones and cytogenetic outcomes in backpack sprayers using ethylenebis(dithiocarbamate) (EBDC) fungicides in Mexico. AB - Ethylenebis(dithiocarbamate) (EBDC) fungicides are used heavily in the United States. EBDCs (e.g., mancozeb, maneb) are metabolized to ethylene thiourea (ETU). The EPA classifies ETU as a carcinogen, based on thyroid and other cancers in rodents, and has restricted the use of EBDCs, while requiring workers to use protective equipment. There are no data on the potential carcinogenicity of EBDCs in humans, and there is only one study on human genotoxicity. ETU is known to cause decreases of thyroxine (T4) and increases in thyroid-stimulating hormone (TSH) in rodents. We have studied cytogenetic outcomes and serum thyroid hormone levels among 49 heavily exposed workers without protective equipment spraying EBDC on tomatoes in Mexico. We also studied 14 lightly exposed landowners and 31 nonexposed controls. Urinary ETU was used to compare exposure between groups. We found an increase in TSH (p = 0.05) among applicators compared to controls, but no decrease in thyroid hormone (T4). We found increases in sister chromatid exchange (p = 0.03) and in chromosome translocations (chromosome aberrations that persist through cell division) for applicators compared to controls (p = 0.05). However, the subset of reciprocal translocations showed a lesser increase (p = 0.24). Our data suggest that EBDCs affect the thyroid gland and the lymphocyte genome among heavily exposed workers. However, our data are limited to subclinical outcomes, are of borderline statistical significance, and should be interpreted with caution. PMID- 9349838 TI - Empirical modeling of an in vitro activity of polychlorinated biphenyl congeners and mixtures. AB - The goal of this research is to predict an in vitro activity of polychlorinated biphenyl (PCB) congeners and their mixtures and to describe the relationship between this activity and chemical structure. The test system used multiple PCB concentrations on each cell culture plate in a repeated measures design, which improved precision for comparing between concentration levels. A weighted regression that accounted for this experimental design feature was used in fitting a nonlinear dose-response exponential model to the PCB concentration activity data from an in vitro test system in which 3H-phorbol ester binding was measured in cerebellar granule cells exposed to different PCB congeners to test for their effects on protein kinase C translocation. The model allowed for the minimum level to be less than control, a common slope, and the estimation of the log of the concentration that produces an activity 50% above the control activity (E50) for 36 congeners and 3 commercial mixtures. Next, a weighted logistic regression using a second order response model in the variables Clortho, Clpara, and Clmeta was used to relate the estimated log E50s to indicators of chemical structure. This model was preferred over models that might seem more mechanistically based because in internal validation, it attained a smaller PRESS statistic (the sum of squares between all observed and predicted observations) than other models. Evidently, this second order model makes more efficient use of parameters than other models considered. Plots of the predictions of the logistic second order response model versus log Kow confirm the usual pattern that congeners with intermediate levels of log Kow are the more active. The data of three commercial mixtures were included in this regression by assuming a common combination index (ratio of observed E50 to predicted E50, assuming dose addition). The logistic model suggests that congeners with one, two, or three chlorine substitutions at the ortho position are more active than other congeners. Also, congeners with log Kow between 5.2 and 6.6 are generally more active. The estimated combination index indicated that the joint action of PCB congeners in the three commercial mixtures was less than dose additive. The error sum of squares was significantly large, which may indicate a lack of fit of the logistic model. Empirical Bayes estimates (EBE) are weighted averages of model predictions and observations of E50s and can be better estimates than the fitted model when there is a lack of fit. The PRESS statistic for the EBE indicated larger prediction error than for the logistic model, but the EBE provided better estimates of commercial mixture E50s based on dose addition. This may indicate that the logistic model is not incorporating all the information in the single congener data needed to predict mixtures. PMID- 9349840 TI - Dietary fatty acids influence the activity and metabolic control of mitochondrial carnitine palmitoyltransferase I in rat heart and skeletal muscle. AB - The fatty acid composition of the diet has been found to influence the activity and sensitivity of mitochondrial carnitine palmitoyltransferase I (CPT I; EC 2.3.1.21) to inhibition by malonyl CoA in rat heart and skeletal muscle. The nutritional state of rats has been shown to have less influence on the activity and metabolic control of mitochondrial CPT I in heart and skeletal muscle tissue than in the liver, a tissue in which CPT I activity and sensitivity to inhibition by malonyl CoA can be shown to be regulated acutely under different nutritional conditions. However, because manipulation of the nutritional state in these previous studies was restricted mainly to examining the effect of starvation, this study was undertaken to determine whether, as in liver, the fatty acid content and composition of the diet can regulate the activity and metabolic control of CPT I in heart and skeletal muscle. Rats were fed for up to 10 wk either a nonpurified low fat diet (30 g fat/kg) or a high fat diet (200 g fat/kg) containing one of the following five oil types: hydrogenated coconut oil (HCO), olive oil (OO), safflower oil (SO), evening primrose oil (EPO) or menhaden (fish) oil (MO). Feeding a diet enriched in MO had the most pronounced effect. Rats fed MO had a significantly greater skeletal muscle CPT I specific activity and tissue capacity, and a lower sensitivity of CPT I to malonyl CoA inhibition compared with rats fed a low fat diet, but the duration of feeding required to modulate this sensitivity was longer than that observed previously for the liver enzyme. Progressively greater sensitivity of heart CPT I to malonyl CoA occurred with feeding duration in all groups. These studies indicate that the fatty acid composition of the diet is involved in the regulation of mitochondrial CPT I activity in heart and skeletal muscle. PMID- 9349842 TI - Dietary L-histidine regulates murine skin levels of trans-urocanic acid, an immune-regulating photoreceptor, with an unanticipated modulation: potential relevance to skin cancer. AB - Solar ultraviolet-B radiation (UVB; 290-320 nm) causes skin cancer and suppresses cell-mediated immunity, preventing the rejection of UV-induced tumors. One mechanism initiating UV suppression involves the trans to cis photoisomerization of urocanic acid (UCA), a histidine derivative found in the stratum corneum. The addition of L-histidine to nonpurified mouse diet has been shown to increase skin trans-UCA levels and sensitivity to UVB immune suppression. Specially formulated L-histidine diets (0.40-64 g/kg) fed to BALB/c mice that were monitored over a 19 wk period resulted in an unexpected modulation of skin trans-UCA. ANOVA revealed a group-time interaction, providing initial evidence that the skin levels of trans-UCA were modulating up and down in all groups except the control group (6.4 g/kg diet). We observed that both high (64 g/kg diet) and low (0.4 g/kg diet) levels of dietary L-histidine resulted in the increase of skin trans-UCA to levels significantly higher than those recorded in the control group. In mice fed these histidine levels, skin trans-UCA increased to between 2.9 and 3.6 nmol/mg skin (64 g/kg diet, over 5 wk; 0.4 g/kg diet, over 8 wk) and then decreased to approximately 1.69 nmol/mg skin, the base-line level (64 g/kg diet, over 11 wk; 0.4 g/kg diet, over 17 wk). The increase in trans-UCA levels in mice with low L histidine intake may be the result of protein malnutrition, consistent with weight loss observed in those mice. The modulation of trans-UCA levels in skin by dietary L-histidine has not been previously described; its role in skin cancer development is under investigation. PMID- 9349841 TI - Low glutamine concentrations induce phenotypical and functional differentiation of U937 myelomonocytic cells. AB - L-Glutamine is the most abundant free amino acid of the human body and is essential for the culture of many cell types. Clinically, reduction of glutamine by administration of glutaminase or the use of glutamine analogs is a common therapy for patients with acute lymphocytic leukemia. In the current study, we investigated the influence of glutamine concentrations on the human myelomonocytic cell line U937. Decreasing the glutamine concentration evoked a reduction in DNA synthesis (R2 = 0.9885, P < 0.0001), increased cell volume (P < 0.01) and the cytoplasm/nuclear ratio, and enhanced the development of vacuoles but did not influence cell viability. Culturing cells in reduced concentrations of glutamine augmented the percentage of cells expressing CD64 (Fc receptor for IgG/FcgammaRI, P < 0.01), CD11b (complement receptor type 3/CR3, P < 0.001) and CD71 (transferrin receptor, P < 0.05). The percentage of U937 cells expressing CD23 (low affinity receptor for IgE/FcepsilonRII) was increased at low concentrations of glutamine at both the protein (P < 0.01) and mRNA levels. The percentage of U937 cells phagocytizing opsonized E. coli (P < 0.001) or latex particles (P < 0.001) was enhanced by lowering the glutamine concentration. In conclusion, reducing glutamine concentration causes differentiation of the cell line U937 along the monocytic pathway. These effects may indicate a mechanistic basis for prior published evidence that glutaminase and glutamine antagonists are effective anti-tumor agents. PMID- 9349843 TI - Dietary selenium increases selenoprotein W levels in rat tissues. AB - Two experiments were conducted to evaluate the influence of dietary selenium (Se) on tissue levels of selenoprotein W (Se-W) in rats. Se dependent glutathione peroxidase (GPX) activity and Se levels were also determined for comparative measurements. In the first experiment, rats were fed a basal diet deficient in Se or supplemented with either 0.1 or 4.0 mg Se (as selenite) per kg diet for 6 wk. Se-W levels were significantly higher in muscle, spleen and testes of rats fed 0.1 mg Se per kg diet compared to those fed the deficient diet (controls), and those fed 4.0 mg Se per kg diet had significantly higher levels in muscle, brain and spleen (P < 0. 05) than those fed 0.1 mg Se per kg diet. No further increases, however, occurred in the tests. There was a significant increase (P < 0.05) of mRNA encoding Se-W in muscle with each increase of dietary Se. In the second experiment rats were fed the basal diet or this diet plus 0.01, 0.03, 0.06, 0.1, 1.0, 2.0 or 4.0 mg Se per kg diet. The levels of Se-W in muscle did not increase (P < 0.05) until 0.06 mg Se per kg diet were fed to rats. A very marked increase (P < 0.05) occurred when 1.0 mg Se per kg diet was fed with no further increases with higher levels. There was a linear increase of Se-W in brain (r = 0.89) and spleen (r = 0.98) with the Se concentration in the diet up to 0.1 mg Se per kg where a plateau was reached. The testes showed a different pattern in that a very marked increase (P < 0.01) occurred when only 0.01 mg Se per kg diet was fed where an inflection was reached. Except for muscle, GPX activities reached a plateau in all tissues when diets containing 0.06 to 0.1 mg supplemental Se per kg were fed. The Se concentration in these tissues increased at a linear rate with the Se concentration in the diets up to 0.1 mg Se per kg where it continued to rise at a different rate. The results indicate that in rats, the regulation of Se-W by Se is different for various tissues and differs from that for GPX. PMID- 9349844 TI - Ascorbic acid deficiency decreases the renal level of kidney fatty acid-binding protein by lowering the alpha2u-globulin gene expression in liver in scurvy-prone ODS rats. AB - The evidence for the role of ascorbic acid in gene expression or protein synthesis in vivo is limited. To investigate this role of ascorbic acid, we surveyed proteins whose tissue levels are changed by ascorbic acid deficiency by using ODS rats with a hereditary defect in ascorbic acid biosynthesis. Male ODS rats (7 wk old, body weight approximately 130 g) were fed a basal diet containing ascorbic acid (300 mg/kg diet) or an ascorbic acid-free diet for 14 d. Ascorbic acid deficiency decreased a renal protein with an apparent molecular mass of 17 kDa. The amino-terminal amino acid sequence of 16 residues of this 17-kDa protein was identical to a kidney fatty acid-binding protein known to be generated by proteolytic degradation of alpha2u-globulin, a major urinary protein of adult male rats. alpha2u-Globulin is synthesized in liver, secreted into blood and excreted into urine, but partially reabsorbed by renal proximal tubules. It exists in kidney in a proteolytically modified form. Ascorbic acid deficiency lowered the renal level of kidney fatty acid-binding protein to 53% (P < 0.05) and lowered the serum level of alpha2u-globulin to 52% (P < 0.05) of the level of the control group, but did not affect the amount of alpha2u-globulin excreted into urine. The hepatic level of alpha2u-globulin mRNA of the ascorbic acid deficient rats was significantly lower (30%) than that of the control rats. These results suggest that in male ODS rats, ascorbic acid deficiency decreases the renal level of kidney fatty acid-binding protein by lowering alpha2u-globulin gene expression in liver. PMID- 9349845 TI - Decreasing traditional food use affects diet quality for adult Dene/Metis in 16 communities of the Canadian Northwest Territories. AB - We assessed diets in 16 Dene/Metis communities in the Canadian Arctic. We described nutrient intakes and identified nutrients at risk among adult Dene/Metis, evaluated the influence of traditional food on diet quality, and examined the direction of dietary change by comparing intergenerational and between-community differences in dietary intake. Diet varied according to sex, age and community. Nutrients of possibly inadequate intake (irrespective of subject sex, age or community) included calcium, vitamin A and folic acid. Dietary fiber intake was also of concern. Traditional food (animals and plants harvested from the local environment) was consumed on 65. 4% of interview days; on those days intakes of iron, zinc and potassium were higher (P < 0.05) and those of sodium, fat, saturated fat and sucrose were lower (P < 0.05) than on days when market food only was consumed. In this population, the shift away from traditional food towards a diet composed exclusively of market food was characterized by an increase (P < 0.05) in absolute energy intake and an increase (P < 0.01) in the relative contributions of carbohydrate (particularly sucrose), fat and saturated fat. This pattern of change calls for initiatives to document the current health status of this population and to prevent potential negative health consequences of dietary change. PMID- 9349846 TI - School-based deworming program yields small improvement in growth of Zanzibari school children after one year. AB - Efficacy trials of antihelminthic therapies conducted in Africa have reported improvements in children's growth, but nutritional evaluations of large-scale deworming programs are lacking. We evaluated the first-year effect on growth of a school-based deworming program in Zanzibar, where growth retardation occurs in school children. Children in four primary schools were given thrice-yearly mebendazole (500 mg) and compared with children in four schools that received twice-yearly mebendazole and children in four non-program schools. Evaluation schools were randomly selected and allocated to control, twice-yearly or thrice yearly deworming. Approximately 1000 children in each program group completed the 1-y follow-up. Children <10 y old gained 0.27 kg more weight (P < 0.05) and 0.13 cm more height (P = 0.20) in the twice-yearly group, and 0. 20 kg more weight (P = 0.07) and 0.30 cm more height (P < 0.01) in the thrice-yearly group, compared with the control group. Children <10 y old with higher heights-for-age at baseline had higher weight and height gains in response to deworming. In children >/=10 y old, overall program effects on height or weight gains were not significant. But in this age range, younger boys had significant improvements in height gain with thrice-yearly deworming, and children with higher heights-for age had greater improvements in weight gain with deworming. We conclude that the deworming program improved the growth of school children, especially children who were younger and less stunted, but the improvements were small. More effective antihelminthic regimens or additional dietary or disease control interventions may be needed to substantially improve the growth of school children in areas such as Zanzibar. PMID- 9349847 TI - Factors associated with anemia in refugee children. AB - A nutrition survey was performed in 1990 among children 6 through 35 mo of age living in Palestinian refugee camps in Syria, Jordan, the West Bank, Gaza Strip and Lebanon. Overall, 67% [95% confidence interval (CI): 66, 68] were anemic (hemoglobin <110 g/L), ranging from 54% in the West Bank to 75% in Syria. The following factors were significantly associated with anemia in one or more of three age groups (6-11.9, 12-23.9 and 24-35.9 mo) by logistic regression: living in Syria, Lebanon, or Gaza [with prevalence odds ratios (POR) in the range of 1.4 2.6 depending on the age group and area, relative to children living in Jordan]; never having been breast-fed (POR = 1.7); male sex (POR = 1.2); maternal illiteracy (POR = 1.4 relative to those with >/=6 y of education); having a recent (within 2 wk) or current episode of fever or diarrhea; and stunting. Recent or current illness and stunting interacted in two age groups with the general trend of stunted children with recent or current illness having high POR. Early childhood anemia is associated with factors reflecting poor socioeconomic status and recent diarrheal and febrile illnesses in Palestinian refugee camps. PMID- 9349848 TI - Nicotinamide counteracts alcohol-induced impairment of hepatic protein metabolism in humans. AB - We have recently shown that a large amount of wine (750 mL, approximately 70 g of alcohol) markedly impairs postprandial hepatic protein metabolism in healthy subjects. This is probably due to the shift in the intracellular redox state (increased NADH/NAD+) induced by ethanol oxidation. If this hypothesis is true, the administration of nicotinamide (NAD+ precursor) should provide NAD+ in excess and thus correct the NADH/NAD+ abnormalities and prevent the ethanol hepatotoxicity. Whole-body protein metabolism and the fractional secretory rates of hepatic (albumin, fibrinogen) and extra-hepatic (immunoglobulin G, IgG) plasma proteins were measured in the basal postabsorptive and in the absorptive states in 15 healthy subjects, that had been assigned to three groups matched for age and body mass index. During the absorptive state (intragastric meal), the three groups received water (control), 750 mL of wine, or 750 mL of wine + 1.25 g of nicotinamide, respectively. The redox state was estimated by determining the plasma lactate/pyruvate ratio. Compared with the basal state, wine alone increased the lactate/pyruvate ratio twofold and depressed the fractional secretory rates of albumin and fibrinogen (P < 0.01 vs. control and nicotinamide); nicotinamide reduced the effects of wine on the lactate/pyruvate ratio (P < 0.02 vs. wine alone) and prevented the reduction of albumin and fibrinogen secretory rates (P > 0.05 vs. control). These results indicate that nicotinamide counteracts the acute hepatotoxic effects of ethanol by ameliorating the redox state. PMID- 9349849 TI - Biotin biotransformation to bisnorbiotin is accelerated by several peroxisome proliferators and steroid hormones in rats. AB - Bisnorbiotin and biotin sulfoxide are the major catabolites of biotin for humans, swine, and rats. Increased urinary excretion of bisnorbiotin, biotin sulfoxide, or both have been observed during pregnancy and in patients treated with certain anticonvulsants. We sought more insight into the sites and mechanisms of biotin catabolism by exposing rats in vivo to compounds known to induce classes of enzymes that were candidates to catalyze the biotransformations. Rats were treated with the anticonvulsants phenytoin, phenobarbital, and carbamazepine, the steroid hormones dexamethasone and dehydroepiandrosterone, and the peroxisome proliferators clofibrate and di(2-ethylhexyl)phthalate. [14C]Biotin was injected intraperitoneally at physiologic doses in treated rats and control rats; HPLC and radiometric flow detection were used to specifically identify and quantify [14C]biotin and its metabolites in urine. Treatment effects were assessed by the change in the urinary excretion of [14C]bisnorbiotin and [14C]biotin sulfoxide in response to administration of [14C]biotin. No significant changes resulted from treatment with any of the anticonvulsants. With the steroid hormones and the peroxisome proliferators, [14C]bisnorbiotin excretion increased significantly. These results indicate that biotin is converted into bisnorbiotin in the liver and that this conversion likely occurs in peroxisomes or mitochondria or both via beta-oxidative cleavage, and, in contrast to responses in humans, the enzymes responsible for the formation of biotin sulfoxide in rats are not induced by the anticonvulsants examined here. PMID- 9349850 TI - Wheat bran diet reduces tumor incidence in a rat model of colon cancer independent of effects on distal luminal butyrate concentrations. AB - To investigate the effects of dietary fibers in colonic luminal physiology and their role in the prevention of colon cancer, a study was conducted using two diet groups and two treatment groups in a 2 x 2 factorial design. The two diets differed only in the type of dietary fiber, wheat bran and oat bran, and the two treatments were injection with the colon-specific carcinogen azoxymethane, or saline, as a control. There were 34 rats in the carcinogen-injected groups and 11 saline-injected rats per diet group. The goal of the study was to determine if a moderate consumption (6 g/100 g diet) of wheat bran or oat bran would alter the development of colonic tumors in this rat model of colon cancer, and if the differences in tumor incidence were correlated to luminal butyrate concentrations, luminal pH or fecal bulk. Short-chain fatty acid concentrations (SCFA) were measured in feces during the first half of the study (the promotion phase of tumor development) and again at the end of the study. Rats consuming oat bran had greater body weights (P < 0. 002), produced much larger concentrations of all SCFA, including butyrate, in both the proximal and distal colon (P < 0.0001), had more acidic luminal pH values (P < 0.0001), but also had significantly more development of colon tumors (P < 0.03). Alternatively, rats consuming wheat bran produced more typical molar ratios of the SCFA (65:10:20), had a relatively greater concentration of butyrate than propionate, and produced a larger volume (P < 0.05) and more bulky stool than the rats fed oat bran. The results of this study support other evidence that an acidic luminal pH is not protective in and of itself, and that diets containing wheat bran are protective against colon cancer development. In addition, these data show that large luminal butyrate concentrations in the distal colon alone, as were present in the rats consuming oat bran diets, are not protective of tumor development. PMID- 9349851 TI - Greater concentration of dietary sucrose decreases dentin formation and increases the area of dentinal caries in growing rats. AB - The effect of increasing dietary sucrose concentration on dentin formation and dentinal caries progression was studied. Weanling Wistar rats received 15, 30 or 43 g/100 g sucrose in a diet; for reference, another group was fed a nonpurified diet. At the onset, tetracycline was injected to mark the dentin formed during the experiment. After 6 wk, lower molars were sectioned sagittally; the areas and thicknesses of the dentin formation during the experiment and dentinal caries lesions were quantified separately in the first and second molars. Feeding the 43% sucrose diet resulted in a significantly lower dentin formation than in other diet groups (P < 0.05). The differences obtained from the area measurements were supported by thickness measurements. In the first molar, the 43% sucrose diet resulted in a significantly greater area of dentinal caries than in the other sucrose groups. The number and severity of caries lesions clearly increased as the concentration of sucrose in the diet increased (r = 0.5, P < 0.05 and r = 0.6, P < 0.05, respectively). This study suggests that the increase in the concentration of sucrose in the diet reduces dentin formation and increases the area of dentinal caries as well as the number and severity of caries lesions; the critical sucrose concentration appears to be between 30 and 43 g/100 g. PMID- 9349852 TI - Phenytoin-induced depletion of folate in rats originates in liver and involves a mechanism that does not discriminate folate form. AB - The anticonvulsant phenytoin causes a decrease in plasma concentrations of folate in epileptic patients. The mechanism underlying this depletion is unknown. To study this mechanism, phenytoin was administered to rats by addition to the diet (3 g phenytoin/kg diet) for up to 8 wk. At selected times during phenytoin administration (0, 3, 7, 10, 14, 28, 42 and 56 d), the composition of the folate pools of intestinal mucosa, liver, bile and brain was determined. The 0-d administration served as the control group. The controls were fed the same diet without phenytoin for the eight weeks of the experiment. Phenytoin administration had minimal effect on either the folate concentration or the composition of the folate pool in intestinal mucosa. Phenytoin administration did, however, cause a depletion of total hepatic folate to about 50% of control, causing the pentaglutamate derivatives of each of the pteridine derivatives to decline rapidly, with the formyl and dihydro derivatives of the pteridine moiety falling more rapidly than the methyl and methylene + tetrahydro derivatives. The monoglutamate of the methylene + unsubstituted tetrahydro derivative increased significantly with time of phenytoin treatment. The mono- and di-glutamate derivatives of the methyltetrahydrofolate increased transiently and significantly in the bile, and the polyglutamate chain length increased significantly in the brain with time of phenytoin treatment. We conclude that phenytoin inhibits the formation of polyglutamyl folates in rat liver. PMID- 9349853 TI - A cholesterol-rich diet causes a greater hypercholesterolemic response in pregnant than in nonpregnant rats and does not modify fetal lipoprotein profile. AB - To determine whether pregnancy modifies the hyperlipidemic response to a cholesterol-rich diet, pregnant and virgin rats were fed a semisynthetic diet supplemented (CRD) or not (CD) with 2% cholesterol and 1% cholic acid and studied at d 20 of treatment and/or gestation. Plasma triglycerides, free fatty acids and glycerol and liver triglycerides were greater in pregnant than in virgin rats fed CRD. The increase in both plasma and liver cholesterol caused by CRD did not differ in the two groups. In rats fed CD, hepatic lipase activity in liver was lower in pregnant than in virgin rats, while in those fed CRD, virgin rats had lower activity than those fed CD. Plasma VLDL-triglycerides were higher and LDL triglycerides lower in pregnant than in virgin rats fed CD. Among those fed CRD, pregnant rats had a higher triglyceride concentration in VLDL and HDL than virgin rats. Cholesterol concentration was higher in VLDL and IDL and lower in HDL in both groups fed CRD than in those fed CD, while cholesterol level in LDL was higher only in pregnant rats fed CRD than in those fed CD. Whereas placental cholesterol concentration was higher in pregnant rats fed CRD than CD, maternal CRD intake did not modify fetal plasma lipoprotein concentrations, fetal body weight or litter size, indicating a lack of cholesterol transfer by the rat placenta. Results therefore show a greater responsiveness to CRD in pregnant than in virgin rats, and we propose that CRD promotes greater liver VLDL-production and lower LDL removal in pregnant than in virgin rats. PMID- 9349854 TI - Soluble non-starch polysaccharides derived from complex food matrices do not increase average lipid droplet size during gastric lipid emulsification in rats. AB - The creation of a finely dispersed lipid emulsion is essential for efficient hydrolysis of dietary triglycerides. The effectiveness of emulsification within the stomach depends upon the shear force generated by gastric motility and the concentration of emulsifiers present in the gastric contents. Other dietary constituents can modify these factors, and previous studies have suggested that the presence of soluble non-starch polysaccharides (NSP) during digestion might increase the average size of intraluminal emulsion droplets. In the present study, we developed a new technique for the isolation and analysis of intraluminal lipid emulsions by optical diffraction analysis. The method was applied to rats fed powdered semipurified diets that were free of all NSP or supplemented with insoluble cellulose, guar gum, or NSP derived from apple, carrot or rolled oats. Cellulose had no significant effect on emulsion size, and there was no evidence that the average sizes of lipid droplets in the gastric fundus or antrum were higher than control values in rats fed diets supplemented with any source of soluble NSP. In the groups fed oats and cooked carrot NSP, the mean droplet diameters approached half the values for diets free of NSP or containing insoluble cellulose. The difference between rats fed NSP from cooked carrot and those fed cellulose was significant in the proximal stomach (P < 0.05), and that between rats fed raw oats and rats fed cellulose was significant in the distal stomach (P < 0.05). Soluble dietary fiber does not inhibit lipid or cholesterol absorption via any inhibition of lipid emulsification. PMID- 9349855 TI - Glutamine supplementation maintains intramuscular glutamine concentrations and normalizes lymphocyte function in infected early weaned pigs. AB - Numerous studies in humans and rats have shown that glutamine supplementation during stressful conditions has favorable outcomes. However, the requirements for glutamine during weaning are unknown. Thus, the effects of glutamine supplementation in healthy and infected weaned pigs were investigated. At 21 d of age, pigs were weaned to an elemental diet supplemented with glutamine (+Gln) or an isonitrogenous diet containing nonessential amino acids (-Gln). At 26 d of age, pigs were intraperitoneally injected with Escherichia coli (+Ecoli) or buffered saline (-Ecoli) and killed at 28 d of age. Infection decreased (P < 0.05) plasma and intramuscular glutamine concentrations, but infected pigs that received +Gln diets had higher intramuscular glutamine levels than those that received -Gln diets. Infected pigs had elevated (P < 0.05) total leukocyte counts, and blood lymphocyte responses ([3H]-thymidine incorporation) to a mixture of phorbol myristate acetate and ionomycin were reduced. White blood cell counts were greater (P < 0.05) in +Gln than -Gln pigs. The peak responses to concanavalin A (Con A) by lymphocytes of +Ecoli+Gln pigs were greater (P < 0.05) than those of +Ecoli-Gln pigs and not different than those of noninfected pigs. Hence, glutamine supplementation maintained muscular glutamine concentrations and normalized lymphocyte function in infected pigs. PMID- 9349857 TI - Dose-dependent effect of betahistine on the vestibulo-ocular reflex: a double blind, placebo controlled study in patients with paroxysmal vertigo. AB - The effect of betahistine on the vestibulo-ocular reflex (VOR) was assessed in 12 patients suffering from paroxysmal vertigo. Only patients who responded to betahistine treatment were admitted to the study in order to increase the probability of quantifying the effect of the drug on vestibular function. Patients received placebo or 16, 32 or 64 mg betahistine orally under double blind conditions. Vestibular function was tested a few minutes before intake, and 1, 2, 3, 4, 6 and 8 h after intake, by torsion swing stimulus in the dark, visuo vestibular interaction upon simultaneous visual and vestibular stimulation and high frequency passive head shaking. Betahistine significantly affected the velocity gain of low and high frequency VOR. The reduction in gain was maximal about 4 h after administration of the 16 mg dose in the torsion swing experiment and the 32 mg dose in the head shaking experiment. Above these doses, the effect on velocity gain was less marked. Betahistine had no effect on visuo-vestibular interaction or nystagmus duration during low frequency torsion. These results suggest that betahistine has a complex action on H3 receptors and that the site of action may be in the vestibular nuclei. PMID- 9349856 TI - The association of yogurt starters with Lactobacillus casei DN 114.001 in fermented milk alters the composition and metabolism of intestinal microflora in germ-free rats and in human flora-associated rats. AB - The aim of this study was to compare the effects of milk and of various fermented milks on the composition and metabolic activities of the intestinal microflora. Groups of eight rats were fed for 6 wk a diet containing 30% nonfermented milk (M), yogurt (Y), milk fermented with Lactobacillus casei (LcFM) or milk fermented with the association of L. casei DN 114.001 and yogurt starters (LcYFM). In the first study, the survival of the lactic acid bacteria from the fermented milks was assessed by bacterial enumeration in feces of germ-free rats (GF rats) fed milk or fermented milks. The metabolic activities of the lactic acid bacteria were studied in these rats by the measurement of glycolytic activities and products of bacterial fermentation, i.e., acetate and lactate (isoforms L and D). In a second study, the effects of fermented milks on the composition and metabolism [gas, glycolytic activities, short-chain fatty acids (SCFA), alcohol and ammonia] of human flora were studied using human flora-associated rats (HF rats). In GF rats, the survival of L. casei in the feces did not differ between those fed the LcFM and LcYFM diets. L. bulgaricus was detected in the feces of the rats fed Y, whereas Streptoccus thermophilus was found in the feces of the LcYFM group. In HF rats, fecal concentration of Bifidobacteria was greater in the LcFM group than in the others. beta-Glucuronidase (EC 3.2.1.31) activity was lower in rats fed LcFM and Y than in those fed M and LcYFM, whereas beta galactosidase (3.2.1.23), alpha-glucosidase (EC 3.2.1 20) and beta-glucosidase (EC 3.2.1.21) activities were higher in the LcYFM group compared with the others. Methane excretion was higher in rats fed Y than in other groups. Cecal SCFA concentrations did not differ in LcFM, Y and M groups, but total SCFA, acetate, propionate and butyrate were significantly greater in the LcYFM group. These results suggest that milk fermented with the combination of L. casei and yogurt starters leads to specific effects that are different from the simple addition of the effects found with yogurt and milk fermented with L. casei. These specific effects are potentially beneficial to human health. PMID- 9349858 TI - Yaw sensory rearrangement alters pitch vestibulo-ocular reflex responses. AB - Ten male subjects underwent two types of adaptation paradigm designed either to enhance or to attenuate the gain of the canal-ocular reflex (COR), before undergoing otolith-ocular reflex (OOR) testing with constant velocity, earth horizontal axis and pitch rotation. The adaptation paradigm paired a 0.2 Hz sinusoidal rotation about an earth vertical axis with a 0.2 Hz optokinetic stimulus that was deliberately mismatched in peak velocity or phase and was designed to produce short-term changes in the COR. Preadaptation and postadaptation OOR tests occurred at a constant velocity of 60 degrees/sec in the dark and produced a modulation component of the slow phase velocity with a frequency of 0.16 Hz due to otolithic stimulation by the sinusoidally changing gravity vector. Of the seven subjects who showed enhancement of the COR gain, six also showed enhancement of the OOR modulation component. Of the seven subjects who showed attenuation of the COR gain, five also showed attenuation of the OOR modulation component. The probability that these two cross-axis adaptation effects would occur by chance is less than 0.02. This suggests that visual vestibular conditioning of the yaw axis COR also induced changes in the pitch axis OOR. We thus postulate that the central nervous system pathways that process horizontal canal yaw stimuli have elements in common with those processing otolithic stimuli about the pitch axis. PMID- 9349859 TI - Vestibulo-ocular reflex function as measured with the head autorotation test. AB - The vestibulo-ocular reflex (VOR) of 125 healthy subjects was examined over the frequency range of 0.5-5 Hz with the head autorotation test (HART). During the HART the subjects fixated at a steady target while moving their heads horizontally from side to side with increasing frequencies according to auditory signals. The gain was determined as the ratio between the amplitude of the eye and head movements in five frequency bands between 0.5 and 5 Hz. The phase difference between the eye and head movements was determined in both degrees and milliseconds. The ability to reach high-frequency bands was evaluated. The mean gain was close to unity up 5 Hz, when it decreased to 0.91. The mean phase difference showed a lead of approximately 5 degrees at frequencies below 2 Hz, and at frequencies above 2 Hz there was no phase difference within the resolution of the test. The frequency band of 5 Hz was reached by 78% of the subjects, and that of 4 Hz was reached by 94% of the subjects. In summary, the HART is a new approach with which to study VOR function and determine accurately the VOR for healthy subjects. The normal upper frequency limit is 4 Hz in the HART. PMID- 9349860 TI - Intraoperative monitoring of facial nerve antidromic potentials during acoustic neuroma surgery. AB - The present paper presents monopolar recording of facial nerve antidromic potentials as an alternative technique to facial electromyography for the continuous monitoring of the facial nerve during acoustic neuroma surgery. The investigation involved 22 patients undergoing acoustic neuroma surgery via a retrosigmoid approach (tumour sizes ranging from 5 to 28 mm). Bipolar electrical stimulation of the marginalis mandibulae was performed to elicit facial nerve antidromic potentials. Stimulus intensity ranged from 2 to 6 mA with a delivery rate of 7/sec. A silver wire monopolar electrode positioned intracranially on the proximal portion of the acoustic facial bundle was used to record antidromic potentials. To define the specific origin of the action potentials and acquire normative data, monopolar and bipolar recordings of facial nerve antidromic potentials were performed in 15 subjects undergoing retrosigmoid vestibular neurectomy for Meniere's disease. The average facial nerve antidromic potential latency was 4.2 (+/- 0.6) msec in subjects with acoustic neuroma and 3.3 (+/- 0.2) msec in subjects with Meniere's disease. Facial nerve antidromic potentials furnished near real-time information about intraoperative facial nerve damage and postoperative facial nerve function during acoustic neuroma surgery. Facial nerve antidromic potentials may provide additional information to conventional EMG. They allow the use of endplate blockers, yield quantitative estimation of facial nerve conduction properties in terms of amplitude and latency, and allow actual continuous monitoring of the facial nerve. PMID- 9349861 TI - Effect of delayed facial-facial nerve suture on facial nerve regeneration. A horseradish peroxidase tracing study in the rat. AB - In clinical practice, the lesioned facial nerve is usually restored by facial facial nerve anastomosis (FFA) with some delay. The optimal time-point for facial nerve reconstruction is still unknown. This study, using rats, compared the effects of immediate and delayed FFA, i.e. FFA 7-56 days after interruption of the facial nerve. Muscle reinnervation was studied 42 days after nerve suture by counting all retrogradely labelled facial motoneurons after injection of horseradish peroxidase (HRP) into the whiskerpad of the rats. Immediate FFA caused a local hyperinnervation of the target muscle, i.e. the projection of more neurons into the whiskerpad muscles than under normal conditions. FFA delayed for 7 days resulted in a significant suppression of this hyperinnervation, whereas longer delay times of 10-56 days showed no difference from immediate FFA. PMID- 9349862 TI - An electrophysiological study of the effects of acute methylmercury chloride exposure on the function of the guinea pig cochlea. AB - The inner ear function of methylmercury chloride (MMC)-exposed guinea-pigs was examined in this study. Previous studies which investigated the function of the eighth cranial nerve and Corti-organ using cochlear microphonics (CM), compound action potential (CAP) and measurement of endocochlear potential (EP) reported ototoxicity following experimental exposure to MMC. In this report, the effect of MMC on the cochlea and the eighth cranial nerve were investigated systematically by measuring CM, action potential (AP), EP and K+ ion concentration of the endolymph. Guinea-pigs were injected with 5 mg/kg MMC (using 0.2% solution) twice a week for 1-3 weeks. The maximum output voltage of AP was decreased by injection of MMC (5 mg/kg x 6). A decrease in the CM maximum output voltage and the elevation of CM pseudothreshold was seen after MMC injection. Changes in EP during 3 min anoxia were observed, especially a decrease in the absolute value of the negative potential. The endolymph K+ ion concentration remained unchanged. These findings indicate that the diffusion potentials decreased and at the same time was reduced the maximum output voltage in CM induced by MMC injection (5 mg/kg x 6) in this experiment. PMID- 9349863 TI - Cochlear changes following destruction of semicircular canal in healthy and previously toxin-exposed rats. An electrophysiological and morphological investigation. AB - One group of Sprague-Dawley rats (group A, n = 6) was treated by instilling Pseudomonas aeruginosa exotoxin A (PaExoA), and another (group B, n = 6) treated similarly with Haemophilus influenzae type b endotoxin (HiBEndo). In group A a 20 dB hearing loss was observed, predominantly in the high-frequency region, which was reversible within 1 month. In group B no significant hearing impairment was noted. Between 1 and 6 months later, the lateral and posterior semicircular canals (SCCs) were ablated unilaterally. Control rats (group C, n = 8) were subjected to ablation only. All rats were cochleotomized contralaterally prior to labyrinthine surgery. Frequency-specific evoked potential testing at 2-31.5 kHz tone bursts was performed before and directly after surgery, 6, 24 and 48 hours and 1, 4 and 16 weeks postoperatively. After surgery in 18 rats, thresholds rose immediately, predominantly at 2, 4 and 6 kHz, followed by varying degrees of recovery. Greatest immediate postoperative hearing loss was observed in group A; no rat recovered completely and two rats showed severe permanent threshold elevation. All group B rats recovered completely, except one showing moderate threshold impairment. No permanent hearing loss was observed in group C. This study shows that destruction of SCCs in rats does not necessarily cause permanent hearing loss, even if the fluid spaces are not sealed off. However, previous exposure of the middle ear to PaExoA (but not HiBEndo) renders the cochlea more vulnerable and can result in persistent hearing loss. PMID- 9349864 TI - Pathology of the cochlea following a spontaneous mutation in DBA/2 mice. AB - The DBA/2 strain of mice usually presents with noise-induced epileptic seizures and hearing disorders. After a spontaneous mutation a strain with early hearing loss and circling behaviour was produced. This strain presents with clinical symptoms found in diseases connected to inner ear disorders. These animals do not suffer from periodical disorders, however, but have functional disturbances continuously and can therefore serve as an animal model for diseases originating from both parts of the inner ear. The genetic inheritance appears to be autosomal recessive. Offspring showed circling behaviour and severe pathology in the vestibular part of the inner ear. In the present study pathology of the cochlear part of the inner ear was visualized using conventional microscopical techniques. The content of actin and fodrin was labelled immunohistochemically, and hearing was assessed with auditory brainstem recordings. After 1 month the animals showed deterioration of the cochlear part of the inner ear. At 6 months no organ of Corti remained and the animals were deaf. Transmission and scanning electron microscopy revealed severe apical hair cell changes. The content of alpha-actinin and fodrin in the DBA/2 mouse was already fainter than that in age-matched CBA control mice at the age of 1 month. Labelling of antibodies against fodrin increased in the supporting cells of the older animals, probably owing to the replacement of hair cells. PMID- 9349865 TI - Otitis-prone children and controls: a study of possible predisposing factors. 2. Physical findings, frequency of illness, allergy, day care and parental smoking. AB - In a retrospective study of 179 otitis-prone children and 305 controls, various possible predisposing factors for acute otitis media (AOM) were compared. The children were matched with the controls for age and sex. There were 61% boys and 39% girls in the otitis-prone group and 58% boys and 42% girls among the controls. Information about the family and living conditions, the children's illnesses, ear, nose and throat (ENT) operations and possible allergies were obtained from a questionnaire, and the children were called for a physical examination. The otitis-prone children had more middle-ear problems with pathological tympanograms and conductive hearing loss than the controls. No differences were found in bacterial colonization of the nasopharynx. Besides AOM and secretory otitis media, the otitis-prone children had more other ENT diseases and had consequently undergone more ENT operations and hospitalizations than the controls. There were no differences between the two groups regarding allergy, day care or parental smoking alone, but on comparing children with combinations of these factors there were more otitis-prone children than controls exposed, indicating an additive effect. The combination of different factors, less important separately, may for some children mean the difference between becoming otitis-prone or not. PMID- 9349866 TI - Histomorphometric study of the normal middle ear mucosa. Preliminary results supporting the gas-exchange function in the postero-superior part of the middle ear cleft. AB - This study concerns the distance between the centre of gravity of blood vessels and the basement membrane of the middle ear cleft mucosa. The measurements were performed perpendicular to the long axis of the cross-section of the vessels, and revealed a significant difference between two regions of the middle ear cleft. At the level of the postero-superior part (epitympanum, aditus ad antrum, mastoid antrum and highest part of the mastoid air cells system), the distance between the blood vessels and the basement membrane of the mucosa was statistically shorter than in the antero-inferior part of the middle ear cleft. This indicates a privileged function of gaseous exchange of the postero-superior part of the middle ear cleft and may divide the middle ear cleft into different functional parts. PMID- 9349867 TI - Increases in middle ear pressure resulting from counter-diffusion of oxygen and carbon dioxide into the middle ear of monkeys. AB - The mechanism of middle ear (ME) pressure regulation is incompletely understood. Previously, Hergels and Magnuson reported an unexpected increase in ME pressure for human subjects who partially evacuated their relative positive ME pressures by swallowing. They suggested that a novel, but unknown mechanism for the generation of gas was responsible for the pressure increase. In this experiment, the MEs of rhesus monkeys were inflated with N2 via the eustachian tube and post inflation ME pressures were recorded for up to three hours. Eleven of 20 experiments showed an initial increase in ME pressure caused by the inflation followed by stable pressures for the remainder of the followup period. In 9 of the experiments, a rapid and temporally discrete decrease in ME pressure was observed during the course of followup. Following the observed pressure decrease which was interpreted as a transient eustachian tube opening, ME pressures showed a progressive increase characterized by a kinetic pattern similar to that of the human experiment. To understand the mechanism responsible for this effect, the monkey experiment was simulated using a mathematical model of ME pressure regulation. Free parameters of the model were taken from experimental data for monkeys. The model accurately predicted the time course of pressure change documented in the monkey experiments. The effect is driven by a decrease in the preexisting ME partial pressures of CO2, O2 and H2O consequent to tubal openings at relative positive pressures, with the subsequent counter diffusion of those gases from blood to ME causing the observed increase in total ME pressure. The results reported by Hergels and Magnuson can also be explained by this mechanism. PMID- 9349868 TI - Keratinocyte growth factor and receptor mRNA expression in cholesteatoma of the middle ear. AB - Keratinocyte growth factor (KGF) is synthesized and secreted exclusively by stromal cells in epithelialized organs, and specifically promotes proliferation of cells of epithelial origin, including keratinocytes. Cholesteatoma is a proliferative form of keratinocyte dysregulation with aggressive growth that often leads to destruction of adjacent bone. We examined the expression of KGF, closely related peptides and their receptors in the development of cholesteatoma by comparing cholesteatoma with normal ear skin using the reverse transcription polymerase chain reaction (RT-PCR) followed by Southern blot analysis. All surgical specimens of cholesteatoma and most normal ear skin samples expressed detectable levels of KGF and KGF receptor (KGFR). Semiquantitative RT-PCR using beta-actin mRNA as an internal standard revealed significantly higher expression of KGF mRNA in cholesteatoma than in normal skin, while no significant difference in KGFR mRNA expression was noted between cholesteatoma tissue and normal skin. These results suggest that excessive KGF synthesis may contribute to the hyperproliferative state in cholesteatoma. PMID- 9349869 TI - In vivo induction and regulation of interleukin-8-like chemokine GRO/CINC-1 in rat middle ear. AB - Interleukin-8 possesses chemotactic-activating properties toward neutrophils, and may contribute to the pathogenesis of middle ear inflammation. GRO/CINC-1 is a rat chemokine with structural and functional homology to human interleukin-8, the induction and regulation of which in the middle ear cavity in vivo remains to be established. The production of GRO/CINC-1 in middle ear lavage and gene expression in the middle mucosa was investigated using topical inoculation with lipopolysaccharide (LPS) in the rat in vivo model. GRO/CINC-1 in middle ear lavage showed time- and dose-dependent production under LPS stimulation. The peak of the GRO/CINC-1 production was reached by 4 h after LPS 1 h exposure, whereas the level of production subsequently returned to the level without LPS stimulation at 8 h after LPS stimulation. The topical corticosteroid perfusion in the middle ear after LPS stimulation significantly reduced the production of GRO/CINC-1 in the middle ear cavity compared with that without corticosteroid. At the time of peak production, the expression of GRO/CINC-1 mRNA, evaluated using the polymerase chain reaction, was considerable in the middle ear mucosa. This investigation of the characteristics of interleukin-8-like cytokine in the middle ear cavity using a rat in vivo model has extended the functional concept of chemokines at the initial stage in otitis media. PMID- 9349870 TI - Nasal congestion in relation to low air exchange rate in schools. Evaluation by acoustic rhinometry. AB - Upper airway symptoms are common, but there is little information available on clinical findings in relation to indoor air pollution. This pilot study was conducted to test whether increased levels of indoor air pollutants in schools may correlate to a swelling of the nasal mucosa. The assumption was made that the degree of swelling could be related to the degree of decongestive effect of xylometazoline, and measured by acoustic rhinometry. The study was performed among 15 subjects in a school with low air exchange rate (0.6 air changes/h) and 12 subjects in a school with high air exchange rate (5.2 air changes/h). Hygienic measurements were performed in both schools. Acoustic rhinometry was performed for each individual under standardized forms. Cross-sectional areas and volumes of the nasal cavity were measured before and after decongestion with xylometazoline hydrochloride. Absolute values of the minimal cross-sectional area were lower in the school with poor ventilation. The decongestive effect of xylometazoline was significantly higher in the school with low air exchange, when correction for the influence of age was made. A diminished decongestive effect was seen with increasing age. The exposure measurements showed that indoor concentrations of volatile organic compounds, bacteria and moulds were higher in the school with low ventilation. In conclusion, raised levels of indoor air pollutants due to inadequate ventilation in schools may affect the upper airways and cause a swelling of the nasal mucosa, and acoustic rhinometry could be a useful objective method to measure human nasal reactions to the indoor environment. PMID- 9349871 TI - Low levels of nasal nitric oxide (NO) correlate to impaired mucociliary function in the upper airways. AB - Findings in previous studies have suggested nitric oxide (NO) to be a regulator of mucociliary activity in the upper airways. The aim of the present investigation was to study whether a correlation exists between the nasal NO concentration and mucociliary function in patients suffering from respiratory tract diseases such as chronic sinusitis or recurrent pneumonia. Nasal NO was measured with a chemiluminescence analyser, 100 ppb (parts per billion) being adopted as the lower limit of the normal range on the basis of findings in an earlier study of healthy subjects. Mucociliary function was evaluated by measurements of ciliary beat frequency (CBF) in nasal brush samples, and the saccharin transport test. A subnormal level of nasal NO was found in 50% (9/18) of the patients. This correlated with a significantly impaired mucociliary function, regarding both CBF and the saccharin transport time. The median CBF was 10.6 Hz in the group with normal levels of nasal NO, as compared to 8.4 Hz in the subnormal NO group. All patients with a normal nasal NO concentration had a mean CBF of > or = 9.0 Hz in their nasal brush samples, but in the subnormal group the same measurements yielded a CBF of > or = 9.0 Hz in only 22% (2/9) of the cases. As measured with the saccharin test, mucociliary transport was normal in 78% (7/9) in the normal nasal NO group, but the saccharin test was normal only in 11% (1/9) of the subnormal nasal NO group. Nasal NO levels were found to correlate with both CBF measurements (Spearman's rho, 0.80) and the saccharin transport test results (Spearman's rho, -0.61). The results of the present study provide further support for the view that NO is an important regulator of mucociliary function in the upper airways, and that measurements of the nasal NO concentration should be included in investigations of the mucociliary system. PMID- 9349872 TI - Distribution and origin of nitric oxide synthase-containing nerve fibers in human nasal mucosa. AB - Human nasal mucosa was investigated by nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. NADPH-diaphorase-positive fibers were distributed around blood vessels and seromucous glands, but none was seen in the respiratory epithelium. The pterygopalatine ganglion was then studied to determine the origin of these fibers, and many neurons were found to be labeled. These labeled neurons (86.2%) consisted of small (14.7%), medium-sized (18.4%) and large cells (53.1%). These results suggest that nitrinergic nerve fibers may originate from the pterygopalatine ganglion and that nitric oxide may play some role in the nervous control of the human nasal mucosa. PMID- 9349873 TI - Localization of 11 beta-hydroxysteroid dehydrogenase: specific protector of the mineralocorticoid receptor in mammalian olfactory mucosa. AB - The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta HSD) plays a major role in the protection of the mineralocorticoid (type I) receptor. The cellular mechanism of aldosterone selectivity relies on the coexpression of mineralocorticoid receptors and 11 beta HSD in the same cells. In the current study, 11 beta HSD activity was localized in the mammalian olfactory mucosa by a histochemical technique which links steroid metabolism with the deposition of formazan. The histochemical reaction results from oxidation of the synthetic substrate 11 beta-hydroxyandrostenedione and is dependent on nicotine-adenine dinucleotide (NAD). This demonstrates the presence of a dehydrogenase activity separate from the nicotineamide-adenine dinucleotide phosphate (NADP)-dependent 11 beta HSD. In the olfactory mucosa, the presence of NAD-dependent 11 beta HSD activity is localized to the sustentacular cells and acinar cells of Bowman's glands. No definite NAD-dependent activity was demonstrated in the olfactory receptor neurons. These data suggest that mineralocorticoid receptors present in acinar cells and sustentacular cells are aldosterone selective. PMID- 9349874 TI - Selective attachment of beta-haemolytic streptococci group A to oropharyngeal epithelium in health and disease. AB - Localization and semiquantification of beta-haemolytic streptococci, Group A (GABHS), GABHS attachment and general bacterial attachment to epithelial cells (bacterial number and morphology) were studied during GABHS-positive acute tonsillitis and pharyngitis infections and among healthy GABHS carriers. Samples were collected from various areas of the oropharynx (palatine tonsils, posterior oropharyngeal wall, palatoglossal arch and buccal mucosa). During acute tonsillitis and pharyngitis, GABHS grew in samples obtained from the palatine tonsils and posterior oropharyngeal wall. The ratio of GABHS colonies to other aerobic colonies increased, and GABHS became attached to epithelial cells of both palatine tonsils and posterior oropharyngeal wall. In GABHS carriers, GABHS were found mainly on the palatine tonsils, but these microorganisms were not attached to the epithelium. Overall bacterial attachment to tonsillar and oropharyngeal epithelial cells increased during active tonsillitis and pharyngitis. PMID- 9349875 TI - Speech perception performance of children with a cochlear implant compared to that of children with conventional hearing aids. I. The "equivalent hearing loss" concept. AB - A new measure has been developed to quantify the speech perception performance of children with a cochlear implant (CI). The method summarizes the speech perception scores obtained on a battery of tests that ranges from very basal tasks up to open speech recognition. The overall performance of a child with a CI on the test battery at a certain time during follow-up is matched to that of a reference group of severely and profoundly hearing-impaired children with conventional hearing aids. This matching procedure results in the expression of the speech perception scores of a child with a CI as an "equivalent hearing loss" value. The equivalent hearing loss concept deals adequately with floor and ceiling effects which inevitably occur when a battery with such a large range of tests is used. To illustrate this, application of the procedure to three children with a CI showed that before implantation, while they were using conventional hearing aids, the equivalent hearing loss was above 120 dB hearing level (HL). At 3 years' follow-up the equivalent hearing loss improved to 70 dB HL in the two children with an aetiology of meningitis. This means that these children were performing as well as children in the reference group with a hearing loss of 70 dB HL. The child with congenital deafness showed minor improvements over time. PMID- 9349876 TI - Speech perception performance of children with a cochlear implant compared to that of children with conventional hearing aids. II. Results of prelingually deaf children. AB - In a previous paper, a method was introduced to transform the results obtained by children with a cochlear implant (CI) on a battery of speech perception tests into an overall value, the "equivalent hearing loss" value. This was achieved by matching the speech perception test scores with those of a reference group of children with conventional hearing aids and hearing loss ranging from 50 to 130 dB hearing level (HL). The equivalent hearing loss values of 16 prelingually deaf children with a CI were plotted as a function of time. There was considerable spread in the rate of progress made by the children in terms of the equivalent hearing loss values. The variables studied, age at onset of deafness/duration of deafness (in the present study, these two factors were indistinguishable) and the communication mode used at the children's school, accounted for 64% of the variance in speech perception performance. A plateau in the performance of the better performers was found which seemed to be caused by the level of hearing (the aided thresholds) with the CI. PMID- 9349877 TI - Vocal tract resonance characteristics of adults with obstructive sleep apnea. AB - Vocal tract acoustic resonance was evaluated in a group of 10 untreated adult males with diagnosed obstructive sleep apnea (OSA) syndrome compared to 10 non OSA adult males. Subjects were required to prolong the vowels /i/, /u/ and /a/, which were subsequently submitted to acoustic analysis of formant frequency and formant bandwidth. Results of the formant frequency analysis indicated lower formant values among the OSA group compared to the non-OSA group, for each vowel type. The lower formant frequencies among the OSA group were attributed to greater vocal tract length compared to non-OSA speakers. The corresponding formant bandwidths for each vowel produced by the OSA group were significantly wider compared to the non-OSA group. The wide formant bandwidths were interpreted to reflect significantly greater vocal tract damping in the OSA subjects, resulting from either excessive vocal tract tissue compliance or general size differences in the length and cross-sectional area of the vocal tract. Discussion focuses on the potential applications of acoustic analysis to aid in the diagnosis and follow-up treatment of OSA. PMID- 9349878 TI - Three-dimensional computer-reconstructed image for studying cancer extension within the hypopharynx. AB - Hypopharyngeal cancer is one of the head and neck cancers with poor prognosis. This is due to the unpredictable extension of the local disease and the high ratio of distant metastasis. To verify the morphological behavior of hypopharyngeal cancer, whole-mount horizontal serial sections were developed using the larynges obtained from total pharyngolaryngoesophagotomy. The technique of processing high-resolution three-dimensional (3D) images of the larynges using Nikon Cosmozone 2SA software is presented. Two representative surgical specimens were used and 3D images reconstructed. In these two particular cases, the cancer infiltrated superiorly through submucosal lymphatic ducts and formed separate daughter nests under the intact mucosal epithelium far above the main tumor. 3D images enhanced the morphological features of these cases. These cases also suggested the difficulty of defining the upper resecting limit in the operation of hypopharyngeal cancer. 3D reconstruction is and will be a crucial modality for studying the morphological behavior of hypopharyngeal cancer. PMID- 9349879 TI - Prognostic factors for swallowing rehabilitation following head and neck cancer surgery. AB - Thirty-two head and neck surgical patients with prolonged moderate to severe aspiration were assessed with videoendoscopic and videofluoroscopic swallowing studies to reveal all components of dysphagia and aspiration. All patients received functional swallowing therapy, and 75% of the patients regained full oral intake diet. The duration of non-oral feeding varied widely. The outcome of swallowing rehabilitation (success or failure, duration of non-oral feeding) was statistically correlated with preoperative tumour stage, patients' age, therapy onset, severity of aspiration and the results of the videofluoroscopic measurements of oral and pharyngeal transit time, pharyngeal delay time, duration of laryngeal closure and cricopharyngeal opening, hyoid and laryngeal elevation, presence or absence of a stenosis at the pharyngoesophageal segment. The following factors proved to be statistically significant for the prognostic estimate of swallowing rehabilitation: preoperative tumour stage, therapy onset, and severity of aspiration. For postoperative swallow recovery, an early therapy onset after thorough diagnostics with videoendoscopic and videofluoroscopic swallowing studies is recommended. Videofluoroscopic measurements will yield some prognostic estimate of oropharyngeal dysphagia and aspiration. Videoendoscopy, by it's availability and immediacy, proved to be useful for monitoring the course of rehabilitation. PMID- 9349880 TI - Establishment and characterization of nine new head and neck cancer cell lines. AB - We established new cell lines from head and neck cancer patients for studies of adhesion molecules and cellular behavior in nine patients with primary or metastatic cancer treated at the Asan Medical Center. Explant cultures of fresh tumor tissue were used to develop new permanent tumor cell lines. Lines were tested for tumor formation and histology in nude mice. Flow cytometry and indirect immunofluorescence were used to assess DNA content and expression of the alpha 6, beta 4, and beta 1 integrin subunits and the intercellular adhesion molecule 1 (ICAM-1). In vitro growth patterns and adhesion to plastic were assessed using phase contrast microscopy. AMC-HN-1 to -8 were derived from patients with squamous cell carcinoma. AMC-HN-9 was from an undifferentiated carcinoma of the parotid gland. The 8 lines we tested produced nude mouse tumors that are identical to the histology of the original tumors. AMC-HN-1, -2, -5, and -9 have epithelioid or spindle cell morphology with poor cell-to-cell and cell-to substrate adhesiveness. AMC-HN-3, -4, -7, and -8 grow as adherent epithelioid monolayers. AMC-HN-6 exhibits multilayer stratification. Four lines are near diploid, 4 are hyperdiploid and 1 is hypodiploid. Only three express ICAM-1. All lines express the alpha 6, beta 4, and beta 1 integrin subunits but to different extent. Four, AMC-HN-1, -2, -5, and -6, express the beta 4 integrin at low levels, AMC-HN-3, -4, -7, and -9, have intermediate beta 4 expression, and AMC-HN 8 has extremely high beta 4 expression. The AMC-HN cell lines are representative in vitro models for the study of head and neck cancer biology. Our preliminary results indicate a close relationship between integrin expression and cell adhesion in vitro. PMID- 9349881 TI - Nebulization of S-carboxymethylcysteine does not adversely affect the mucociliary system in the paranasal sinus and trachea of the healthy rabbit. AB - Chronic sinusitis is a persistent inflammatory impairment of the paranasal sinus. Disturbance of the mucociliary function in the paranasal sinus is the most common finding in chronic sinusitis. S-carboxymethylcysteine (S-CMC) has been shown to directly enhance the ciliary activity of the chronic sinusitis mucosa. Direct contact of the disturbed cilia with S-CMC may recover the reduced beating activity of cilia in chronic sinusitis and the mucosal pathology of the disease can thus be improved. Before S-CMC as medicine for nebulization in the treatment of chronic sinusitis can be clinically applied, however, it should be experimentally established whether nebulization of S-CMC has any adverse effects on the mucociliary system of the respiratory mucosa. The present study was designed to experimentally examine the safety of nebulization of S-CMC especially with regard to the respiratory mucosa. Rabbits were treated with nebulization of three different concentrations of S-CMC solution for 20 min a day for 14 successive days, and their mucosal pathology of the sinus and trachea was examined and compared with that of healthy animals. Nebulization of concentrations of 0.5-10% of S-CMC solution did not affect the ciliary activity in the sinus and tracheal mucosa, nor did this treatment induce pathological changes such as epithelial injury and inflammatory cell accumulation. It is therefore concluded that concentrations of 0.5-10% S-CMC solution are quite safe for the use of nebulization in the treatment of chronic sinusitis. PMID- 9349882 TI - Improvement of mucosal pathology of the sinuses after exposure to sulfur dioxide by nebulization of S-carboxymethylcysteine. AB - Since s-carboxymethylcysteine (S-CMC) can directly enhance the ciliary activity in the maxillary sinus mucosa of patients with chronic sinusitis in the absence of significant organic changes of ciliated cells, the nebulization therapy using this medicine might be more effective in the treatment of chronic sinusitis than oral administration of the medicine. The safety of using 0.5-10% of S-SMC as a medicine for nebulization has been experimentally established. The present study was designed to experimentally examine the effectiveness of nebulization using 0.5-10% of S-CMC solution in the treatment of experimental chronic sinusitis in rabbits recurrently exposed to 20 ppm of sulfur dioxide. Thirty-three healthy rabbits were used; 3 of them were used as controls. The remaining 30 were exposed to 20 ppm of sulfur dioxide for 4 h a day for 4 successive weeks. Twelve animals were not treated with any medication during the post-exposure period, and sacrificed at 24 h or 15 days after completion of the final exposure to sulfur dioxide. The remaining 18 animals were treated with nebulization using 10%, 5% or 0.5% of S-CMC solution for 20 min a day for 14 successive days after the final exposure to sulfur dioxide, and they were sacrificed at 24 h after the final nebulization using S-CMC. At the time of sacrifice, the ciliary activity and the morphology of the sinus mucosa were observed to assess the effectiveness of S-CMC nebulization. In the animals sacrificed 24 h after the final exposure, the mucosa of the sinus demonstrated marked epithelial cell injuries, and the ciliary activity was extremely reduced. Complete recovery of the epithelium and the ciliary activity was not recognized in the animals sacrificed 15 days after completion of the exposure. By contrast, epithelial recovery was more accelerated in the animals treated with S-CMC nebulization during the 14 days after the exposure. In the animals treated with 0.5% of S-CMC, the ciliary activity was inferior to that of the control animals, and the epithelial repair was not complete. In the animals treated with 10% of S-CMC, however, ciliary activity and epithelial morphology were completely recovered. In conclusion, our study suggests that clinical application of 10% of S-CMC nebulization may provide otolaryngologists with a new tool in the treatment of sinus diseases such as chronic sinusitis. PMID- 9349883 TI - The herbal medicine, sairei-to, enhances the mucociliary activity of the tubotympanum in the healthy guinea pig. AB - Mucociliary dysfunction in the tubotympanum is deeply reflected in the clinical manifestations of otitis media with effusion (OME), and clinical application of pharmacological agents with ciliostimulatory action might therefore enhance the mucociliary clearance function of the tubotympanum to more effectively eliminate middle ear effusions to the pharynx. A herbal medicine, sairei-to, enhances the in vitro ciliary activity of the middle ear during culture. However, this ciliostimulatory effect is not always applicable to the mucociliary system in situ, which may be deteriorated following oral administration of sairei-to. The present study therefore aimed at investigating the in vivo effect of sairei-to on the mucociliary system in the tubotympanum of the guinea pig. Thirty healthy guinea pigs were used. Ten animals were treated with oral administration of physiologic saline solution for 14 successive days. The remaining animals were treated with oral administration of 120 or 600 mg/kg body weight of sairei-to for 14 successive days. Each animal was used for examination of the ciliary activity and mucociliary clearance time of the tubotympanum, 24 h after the final treatment. No significant changes in either ciliary activity or mucociliary clearance time of the tubotympanum were observed upon administration of 120 mg/kg of sairei-to, which was equivalent to the clinical human dosage. By contrast, oral administration of 600 mg/kg of sairei-to significantly enhanced the ciliary activity, but failed to significantly accelerate mucociliary clearance in the tubotympanum, although the mean value of the clearance time became shorter. Therefore, our results suggest that sairei-to to some extent stimulates the function of the mucociliary system. In conclusion, the herbal medicine, sairei to, might be useful in the treatment of OME, and preventive administration of this drug may be a new therapy in the treatment of recurrent OME. PMID- 9349884 TI - The herbal medicine, sairei-to, prevents endotoxin-induced otitis media with effusion in the guinea pig. AB - With current pharmacotherapy, otitis media with effusion (OME) is often recurrent and even develops to become chronic. There is now considerable experimental and clinical evidence that the cilia in the tubotympanum play an important part in the prevention of OME. A herbal medicine, sairei-to, has been shown to stimulate the ciliary activity in vitro, and oral administration of the medicine also stimulated the ciliary activity in the tubotympanum rather than physiological states. This study was designed to investigate whether oral administration of sairei-to could prevent experimental OME in the guinea pig. A total of 120 guinea pigs were used. The control group was treated with intratympanic injection of 0.1 ml of physiologic saline solution. The saline-control group was treated with oral administration of physiologic saline solution for 14 successive days. The low dosage group and the high-dosage group were treated with oral administration of 120 and 600 mg/kg of sairei-to for 14 successive days, respectively. The saline control group, the low-dosage group and the high-dosage group were then treated with intratympanic injection of 0.1 ml of lipopolysaccharide solution (100 micrograms/ml) derived from Klebsiella pneumoniae. All 10 animals from the 4 groups were sacrificed 1, 3, and 7 days after the intratympanic injection, to examine ciliary activity, mucociliary clearance time, and mucosal pathology of the tubotympanum. The saline-control group exhibited middle ear effusions and pathologies similar to human OME. The incidence of middle ear effusions in the low-dosage and the high-dosage groups was somewhat reduced compared with the saline-control group. The ciliary activity in the tubotympanum was significantly reduced in the saline-control and low-dosage groups compared with the normal control group. By contrast, the magnitude of reduction in ciliary activity was much smaller in the high-dosage group. The ciliary activity especially in the Eustachian tube and the middle ear close to the tympanic orifice at 3 and 7 days in the high-dosage group was not significantly different from that in the normal control group. Mucociliary clearance time in the high-dosage group was not different from that in the normal-control group throughout the observation period. The groups treated with sairei-to, especially the high-dosage group, exhibited much milder pathological changes in the tubotympanum than did the saline-control group. In conclusion, clinical application of sairei-to could be an effective measure to prevent the occurrence of OME and also the recurrence of the disease, especially OME-prone individuals. PMID- 9349885 TI - Roxythromycin stimulates the mucociliary activity of the Eustachian tube and modulates neutrophil activity in the healthy guinea pig. AB - Low-dosage, long-term erythromycin chemotherapy is useful in the treatment of chronic airway inflammatory diseases such as chronic sinusitis. The exact working mechanism of macrolides behind the clinical effectiveness still remains unclear. However, some have been considered including anti-inflammatory effect, effects on airway secretory functions, and steroid sparing effects. Otitis media with effusion (OME) is a chronic inflammatory disease in the tubotympanum. The epithelium of the tubotympanum is a modified respiratory epithelium. Therefore, macrolides might be effective in the treatment of patients with chronic OME. It was recently demonstrated that macrolides such as roxythromycin (RXM), enhance the ciliary activity in vitro. However, such ciliostimulatory effects found in an in vitro system are not always applicable to the mucociliary system in situ. The mucociliary system in situ might behave differently when in contact with RXM, and might deteriorate following oral administration of RXM. The present study aimed at investigating the in vivo effect of RXM on the mucociliary system in the tubotympanum and neutrophil activities of the guinea pig. The healthy guinea pigs were treated with oral administration of 5 or 50 mg/kg/day of RXM for 14 successive days, and the ciliary activity and the mucociliary clearance time of the tubotympanum was determined 24 h after the final administration. The ciliary activity in the Eustachian tube was significantly increased by oral administration of either dosage of RXM for 14 successive days. In addition, the mucociliary clearance velocity was accelerated by oral administration of such dosages of RXM. Therefore, the present data clearly demonstrate that RXM is a pharmacological agent with ciliostimulation and concurrent acceleration ability of the mucociliary clearance in the Eustachian tube. Oral administration of 5 as well as 50 mg/kg/day of RXM for 14 successive days did not significantly affect the phagocytosis activity of neutrophils but significantly increased the superoxide production of neutrophils. Since the effusions are considered to be the result of inflammatory events in the tubotympanum, the increased superoxide production activity of neutrophils confirmed in our study should result in an increased killing activity of infectious pathogens, thereby leading to control of the disease chronicity. In conclusion, our study argues in favor of the clinical usefulness of RXM in the treatment of OME. PMID- 9349886 TI - Roxythromycin prevents endotoxin-induced otitis media with effusion in the guinea pig. AB - Pharmacological agents that can normalize or enhance the ciliary and mucociliary activity of the tubotympanum should also be able to break the vicious circle of chronic otitis media with effusion (OME). Roxythromycin (RXM) has been shown to enhance the ciliary activity in vitro and also stimulate the mucociliary activity in vivo and may therefore, when clinically applied, prevent not only occurrence but also recurrence of clinical OME. The present study was designed to discuss the possible preventive effect of RXM on endotoxin-induced OME in the guinea pig. A total of 120 guinea pigs were used. The normal control group was treated with intratympanic injection of 0.1 ml of physiologic saline solution. The saline control group was treated with oral administration of physiologic saline solution for 14 successive days. The low-dosage group and the high-dosage group were treated with oral administration of 5 and 50 mg/kg of sairei-to for 14 successive days, respectively. Then, the saline-control group, the low-dosage group and the high-dosage group were treated with intratympanic injection of 0.1 ml of lipopolysaccharide solution (100 micrograms/ml) derived from Klebsiella pneumoniae. All 10 animals in the four groups were sacrificed 1, 3, and 7 days after the intratympanic injection, to examine ciliary activity, mucociliary clearance time, and mucosal pathology of the tubotympanum. The saline-control group exhibited middle ear effusions and pathologies similar to human OME. The incidence of middle ear effusions was significantly reduced in the low-dosage and high-dosage groups. Throughout the observation period, the ciliary activity in the tubotympanum was significantly reduced in the saline-control group as compared with that of the normal control group. By contrast, the ciliary activity in the low-dosage and high-dosage groups was not so reduced in the Eustachian tube and the middle ear close to the orifice. Mucociliary clearance time in the low-dosage and high-dosage groups was not different from that in the normal control group throughout the period. The tubotympanum in the saline-control group exhibited mucosal pathologies characteristic of OME in human. By contrast, the low-dosage and high-dosage groups exhibited much milder pathological changes in the tubotympanum than those in the saline-control group. In conclusion, clinical application of RXM could be an effective measure to prevent the occurrence of OME and also the recurrence of the disease, especially in OME-prone individuals. PMID- 9349887 TI - Bioresorbable fracture fixation in orthopedics: a comprehensive review. Part I. Basic science and preclinical studies. AB - Metal alloys are currently the most popular materials for manufacture of fracture fixation devices. Two major disadvantages of these materials are their extreme stiffness, which causes stress shielding of the underlying bone, and the necessity, in a significant number of cases, of removing metallic implants after fracture healing is complete. These shortcomings of metal alloys have led to the study of bioresorbable materials for use in fracture fixation. Currently, polylactic acid, polyglycolic acid, and polydioxanone implants are available to the orthopedic surgeon for the fixation of small cancellous bone fractures. Part I of this article provides an overview of the basic science of bioresorbable materials and presents a comprehensive review of preclinical studies reported in the orthopedic literature. Clinical studies will be reviewed in Part II. PMID- 9349888 TI - Trigger finger: the effect of partial release of the first annular pulley on triggering. AB - A prospective, randomized study was performed on 19 patients, in whom the proximal, middle, or distal third of the first annular pulley was divided to determine if a specific portion of the first annular pulley was responsible for the clinical triggering associated with restrictive flexor tenosynovitis. In all 19 patients, a partial resection of the first annular pulley resulted in continued clinical triggering with active digital flexion. At this point, a standard complete first annular pulley release was performed, with resolution of clinical triggering of the involved digit in all patients. We conclude that there is no "critical third" of the first annular pulley responsible for clinical digital triggering. PMID- 9349890 TI - Multiple stress fractures of the tibia in a healthy adult. AB - Multiple stress fractures in the same bone are a rare occurrence. This paper presents an unusual case of a 19-year-old, healthy man who sustained a pathologic fracture of the proximal tibia through a stress-fracture site. The radiographic examination of the tibia also revealed multiple stress fractures inferior to the pathologic fracture. Magnetic resonance imaging can be used to rule out neoplasm and to confirm the stress-fracture diagnosis. Multiple stress fractures in the same bone can be treated with cast immobilization. PMID- 9349889 TI - The outcome of Austin-Moore hemiarthroplasty for fracture of the femoral neck. AB - One hundred and fifty-four consecutive patients who sustained a fracture of the femoral neck and were treated with an Austin-Moore hemiarthroplasty were followed up longitudinally for 10 years, or until revision of the prosthesis, or death of the patient. Clinical assessments were performed at 1, 3, 5, and 10 years after surgery. At 3 years, 46% of the patients were community ambulators; 10%, household ambulators; 6%, nonfunctional ambulators; 29%, nonambulators; and 9% unknown. At 10 years, 31% were community ambulators; 35%, household ambulators; 4%, nonfunctional ambulators; and 30%, nonambulators. Men had statistically significantly better ambulation than women at the 3-year review (P = 0.004), but there was no difference at later assessments. The Harris hip score was 69 (range, 41 to 97) at 5 years and 69 (range, 25 to 90) at 10 years. The overall failure rate was 6.5% at 5 years and 7.7% at 10 years. The revision rate was 4.5% at 5 years and 5.2% at 10 years. Patients younger than 70 years at the time of fracture had a statistically significantly higher revision rate than did older patients. PMID- 9349891 TI - Protrusio acetabuli and bilateral basicervical femoral neck fractures in a patient with Marfan syndrome. AB - A 22-year-old man with Marfan syndrome and bilateral protrusio acetabuli presented with bilateral femoral neck stress fractures after vigorous stretching exercises for hip "stiffness." Fifteen years later, his fractures, which were treated with internal fixation, have healed, his acetabular protrusion has not worsened, and his perceived hip "stiffness" persists. This case demonstrates a rare manifestation of Marfan syndrome, protrusio acetabuli, and a possible side effect of vigorous stretching in the face of abnormal joint mechanics. PMID- 9349892 TI - Aneurysmal bone cyst arising after anterior cruciate ligament rupture. AB - An aneurysmal bone cyst, a condition first described in 1942, is a relatively rare benign tumor. Approximately 1% of all primary bone tumors are aneurysmal cysts. Diagnosis is typically made in a patient's first or second decade and rarely follows trauma. This is the first reported case of an aneurysmal bone cyst arising after rupture of the anterior cruciate ligament. PMID- 9349893 TI - Compression neuropathy of the peroneal nerve caused by a ganglion. AB - Peroneal nerve injury usually results from fracture of the fibula neck or direct pressure from an ill-fitting plaster cast. Clinically, these patients present with "drop foot." Compression neuropathy caused by a ganglion is rarely encountered. This report concerns the clinical and radiologic evaluation of a 57 year-old man admitted to our clinic with left drop foot. A mass compressing the peroneal nerve at the level of the fibula neck was excised totally and diagnosed as a ganglion. PMID- 9349894 TI - Alcohol and drug use in adult patients with musculoskeletal injuries. AB - This study reviewed trauma registry data for information on the prevalence of alcohol and drug use in adult patients with fractures and dislocations admitted to Hermann Hospital, Houston, Texas, from January 1992 to December 1994. Of the 1776 adult patients aged 18 years or older, 1126 (63%) were tested for blood alcohol concentration, and 873 (49%) had their urine screened for a panel of 58 drugs. Of the patients tested, 467 (41%) had positive blood alcohol concentrations, and 335 (30%) were legally intoxicated (blood alcohol concentration > or = 0.10%). Of the patients providing urine specimens, 191 (22%) had samples that were positive for one or more drugs. Overall, 9% of the patients tested were positive for both alcohol and drugs, and 54% were positive for either alcohol or drugs. The highest prevalence of alcohol use was in patients aged 21 to 33 years, and men were intoxicated more often than women. Alcohol use was more commonly associated with motor vehicle accidents, pedestrian or bicycle accidents, and gunshot wounds; intoxification was associated with a higher incidence of tibia fractures. The average injury severity score was higher, the duration of stay was longer, and total hospital charges were higher for the alcohol-positive group. Patients testing positive for alcohol or drugs were more likely to lack insurance coverage. PMID- 9349895 TI - A 3-month-old baby with multiple fractures. AB - This case is presented to illustrate the radiographic and clinical findings of a condition of interest to orthopedic surgeons. The initial findings are noted on this page. The clinical and radiographic diagnoses are presented on the following pages. PMID- 9349896 TI - Computer-assisted discrimination of glioblastomas. AB - OBJECTIVE: To measure a number of nuclear features in a series of glioblastomas and compare the data with those from a set of anaplastic and low grade astrocytomas. STUDY DESIGN: The material consisted of toluidine blue-stained smears from 13 consecutive cases of glioblastoma. Smears from 12 high grade astrocytomas and 13 low grade fibrillary astrocytomas were used for comparison. Fifty nuclei were measured in each case. Cell images were segmented by an interactive procedure. A set of features representing both morphometric and nuclear texture characteristics was computed. RESULTS: The use of a discriminant function based on two features related to the gray value distribution resulted in the separation of all low grade astrocytomas from glioblastomas. When the corresponding discriminant function was computed for high grade astrocytomas and the values were plotted against optical density, the glioblastomas formed a group at a greater distance from the low grade astrocytomas than from the high grade astrocytomas. A second discriminant function based on two features allowed complete separation of the glioblastoma cases from the high grade astrocytomas. CONCLUSION: In our material, chromatin texture analysis allowed effective separation of astrocytic tumors with different histologic grades of malignancy. PMID- 9349897 TI - Possibility of correcting image cytometric nuclear DNA (ploidy) measurements in tissue sections. Insights from computed corpuscle sectioning and the reference curve method. AB - OBJECTIVE: To describe, validate and apply a computer simulation of corpuscle sectioning to determine which assumptions are compatible with the possibility of correction. STUDY DESIGN: A computer program was written in PASCAL and run on a personal computer. RESULTS: Correction is practical in the case of spherical nuclei with homogeneous intranuclear DNA and a constant internuclear DNA concentration, even if section thickness is uneven or inaccurate. Correction is not possible in the case of marked nuclear ellipticity: mild cases are correctable, although subpopulations in a mixed-ploidy sample would not be distinguished. Correction is possible in the case of spherical (but not ellipsoidal) nuclei with a variable internuclear DNA concentration, and subpopulations in a mixed-ploidy sample would be distinguishable if sufficiently different. If the DNA is markedly concentrated in one part of the nucleus, overcorrection results. CONCLUSION: The results clarify the factors that limit correction of image cytometric measurements of ploidy in tissue sections. A new method of correcting ploidy measurements in tissue sections is usable. PMID- 9349898 TI - Computer-assisted analysis of medulloblastoma. A cytologic study. AB - OBJECTIVE: To explore data from a set of cases of medulloblastoma to see whether quantitative image analysis might suggest evidence for the existence of lower and higher grade lesions. STUDY DESIGN: Fourteen consecutive cases of medulloblastoma were obtained. Smears were stained with toluidine blue. For each case, 50 nuclei were measured and a number of densitometric features extracted. RESULTS: The existence of two subgroups of cases, identified as lower and higher grade groups, was suggested by a plot of the total optical density versus nuclear area. Two nuclear texture features--the number of pixels with the same optical density value occurring consecutively in the nucleus and the proportion of pixels in the high optical density range--divided the cases into the same subgroups. The use of a clustering algorithm established two clusters that corresponded to that subgrouping except for one case. Discriminant analysis gave an identical classification, with the misplaced case having a borderline discriminant function score. An unsupervised learning algorithm based on an adaptive distance metric formed two clusters and assigned the borderline case to the low grade subgroup. The grouping obtained by quantitative analysis was only partly related to the grade of nuclear atypia subjectively evaluated. CONCLUSION: In our series of medulloblastomas, quantitative analysis provided a means of detecting differences in the nuclear size and texture that allowed the classification of cases into two subgroups. PMID- 9349899 TI - P90 exceeding rate as a prognostic factor in primary malignant melanoma of the skin. A DNA image cytometric study. AB - OBJECTIVE: To study the prognostic value of DNA image cytometry in primary skin melanomas. STUDY DESIGN: DNA image cytometry was performed on 62 stage I, Clark level II-V, primary skin melanomas. The DNA histograms were classified into three categories (diploid, nondiploid and aneuploid) according to the percentages of cells with higher-than-diploid and higher-than-twice-the-diploid DNA content (the P90 and 2P90 exceeding rates [ERs]). The prognostic value of P90ER, 2P90ER, type of DNA histogram, melanoma thickness, Clark level, and patient age and sex were analyzed for disease-specific survival with Cox's stepwise proportional hazards model. RESULTS: Aneuploid DNA histograms were as common in thin as in thick melanomas. Melanoma thickness and P90ER had prognostic value in univariate analysis, but in the multivariate analysis only P90ER had independent and significant prognostic value. CONCLUSION: Aneuploidy is a common feature of malignant melanoma, and it is as common in thin as in thick melanomas. P90ER has more prognostic value than the type of DNA histogram. The prognostic value of P90ER as compared with melanoma thickness should be studied further. PMID- 9349900 TI - Confocal fluorescence analysis of the depth and focus of cytogenetic preparations. AB - OBJECTIVE: To characterize differences in the depth of fluorescent probes, to observe estimated depth levels (focal planes) on fluorescent in situ hybridization preparations by factor analysis of medical image sequences and to use cytogenetic techniques resulting in flat preparations of whole cells that are assumed to preserve the probes' access to their targets in the human nuclear interphase. STUDY DESIGN: We used labeling of the targets by the probes (sequences labeled by fluoroscein isothiocyonate [FITC]) in the nuclei, stained by propidium iodide. The investigation was performed on this model by three dimensional (3-D) sequences of images obtained on a single photomultiplier detector of a confocal microscope by selection of emission between 510 and 550 nm and by (z) displacement. The investigation was also performed by obtaining sequences of images from spherical fluorescent beads to verify (z) focusing, to visualize depth differences and to analyze estimated factor images. RESULTS: Estimated images showed depth differences in FITC-stained targets as well as in nuclei, stained with propidium iodide, in interphase and in fluorescent beads, that could not be visualized by conventional 3-D reconstruction. CONCLUSION: It is possible to study 3-D architecture of flattened preparations and penetration of fluorophores inside the beads and to evaluate depth differences. PMID- 9349901 TI - DNA ploidy analysis in breast carcinoma. Comparison of unfixed and fixed tissue analyzed by image and flow cytometry. AB - OBJECTIVE: To form a methodologic basis for DNA analysis of ductal carcinoma in situ (DCIS) and invasive carcinoma (IC) of the breast, including very small lesions, by comparison of flow cytometric (FCM) and image cytometric (ICM) methods for DNA quantitation. STUDY DESIGN: The material consisted of 41 DCIS lesions and 26 ICs. FCM DNA analysis of unfixed, frozen samples were compared to (1) FCM of formalin-fixed, paraffin-embedded tissue; (2) ICM of imprints; and (3) ICM of paraffin-embedded tissue sections. RESULTS: FCM of unfixed tissue showed higher DNA measurement precision and a higher number of DNA nondiploid clones as compared to the other three methods. For the classification of DNA diploid/nondiploid cases, high concordance rates were found between the methods. Discordant cases were predominantly DNA neardiploid by FCM of unfixed tissue but DNA diploid by the other methods. The reproducibility of the DNA index (DI) was best in the interval 1.2 < DI < or = 2.2; it was 74% for FCM of fixed tissue and 79% for ICM of imprints. Clones with DI > 3 were found almost exclusively by ICM of imprints. For ICM of tissue sections, DI could not be reliably estimated. By ICM, contrary to FCM, a combined DNA diploid and nondiploid pattern was found frequently. CONCLUSION: Each of the methods has its own advantages and limitations. If possible, FCM should be combined with ICM. FCM of unfixed tissue is superior to the other methods with respect to precise DI estimation. Alternatively, FCM of fixed tissue and ICM of imprints may both give a reliable estimate of DI. ICM of tissue sections can discriminate DNA diploid from nondiploid clones, except for neardiploid subpopulations, and permits the analysis of very small lesions. PMID- 9349902 TI - Subjective breast cancer grading. Analyses of reproducibility after application of Bayesian belief networks. AB - OBJECTIVE: To examine the influence of Bayesian belief networks (BBNs) on the reproducibility of subjective breast cancer grading. STUDY DESIGN: Twenty samples were analyzed for intraobserver and 128 samples for interobserver reproducibility using the Bloom-Richardson and Helpap grading systems. The expression of diagnostic features was evaluated subjectively, and for each a decision it was determined to what extent it represented one of the different outcomes. Evidence was then entered, for each diagnostic feature, into four different BBNs, recently described for breast cancer grading, in the form of a relative likelihood ratio vector. RESULTS: With all cases considered, the use of decision support based on the Bloom-Richardson and Helpap grading systems did not improve intraobserver reproducibility. This was found to be 68% and 80% in subjective gradings, respectively, and 60% and 70% in the BBN-supported method. Interobserver reproducibility was not improved (58% and 70% in subjective gradings and 51-59% based on assessment with decision support). However, when only cases associated with high beliefs were considered, both intraobserver reproducibility (agreement rose from 68% to 93%) and interobserver reproducibility (agreement rose from 60% to 87%) of BBN-supported gradings exceeded the results of subjective assessments. CONCLUSION: The results showed that the observers did not reach the same diagnosis (or grade) and that their observational assessment of histologic features lacked agreement. Since BBNs reflected only the data entered, poor agreement existed in the contribution to the final diagnostic belief by the different features and, ultimately, in belief in the final decision. PMID- 9349903 TI - Comparative analysis of DNA content in polyomavirus-infected urothelial cells, urothelial dysplasia and high grade transitional cell carcinoma. AB - OBJECTIVE: To characterize the DNA content pattern in cytologically confirmed polyomavirus (PV)-infected urothelial cells and to compare it with DNA ploidy changes in cytologically diagnosed urothelial dysplasia and high grade transitional cell carcinoma. STUDY DESIGN: We selected 200 bladder cytology specimens consisting of four groups with 50 patients each in the following cytologic categories: (1) no evidence of malignancy or dysplasia (controls), (2) PV, (3) urothelial dysplasia (UD), and (4) high grade transitional cell carcinoma (TCC-HG). For each case, two slides with 25-mm filter imprints were stained, one using the Papanicolaou method and the other using the Feulgen staining method. The DNA index (DI), proliferative activity (S + G2M) and degree of hyperploidy (> 5C) were evaluated using an image analysis system. RESULTS: Using the Wilcoxon rank-sum test, statistically significant differences in the DI were found between the PV and UD groups (P = .008) and between the PV and TCC-HG groups (P < .0001). There was no significant difference in the DI between the PV and control groups. The S + G2M fraction for the PV group significantly differed from the control, UD and TCC-HG groups (all P < .0001). Between all four groups, the degrees of hyperploidy were significantly different as well (all P < .0001). CONCLUSION: Cytologically confirmed PV-infected urothelial cells demonstrated a unique DNA content pattern with mildly elevated proliferative activity and a significantly dispersed hyperploid DNA content pattern. DNA analysis can help to differentiate PV infection from dysplasia and high grade carcinoma. PMID- 9349904 TI - DNA ploidy by image cytometry in urothelial carcinomas. Comparison of touch imprints and paraffin-embedded biopsies from 31 patients. AB - OBJECTIVE: To compare DNA content measured by image cytometry from touch imprints and formalinfixed, paraffin-embedded samples in bladder carcinomas. STUDY DESIGN: Thirty-one biopsies of urothelial carcinomas were selected for a prospective study. Imprints of fresh specimens were performed. Cell suspensions were obtained from dewaxed samples by the procedure of Hedley. Sections 7 microns thick were used for carcinoma in situ and small biopsies. The DNA ploidy index was measured on Feulgen-stained slides using an image cytometer. RESULTS: From imprint analysis, seven grade 1 carcinomas (n = 9) were found to be diploid (78%). Nine grade 2 carcinomas (n = 12) exhibited aneuploidy (75%), as did all grade 3 and in situ carcinomas (n = 10). Multiploidy was demonstrated from imprints in four cases instead of the two detected from dewaxed tissue. In 27 cases (87%), G0/G1 peaks obtained from paraffin blocks showed a shift to the left. In five cases (16%), variations in the DNA index were responsible for discrepancies in the DNA ploidy evaluation between fresh imprints and dewaxed samples of the same tumors. CONCLUSION: Image cytometry on Feulgen-stained imprints of bladder biopsies is a simple and reliable procedure for assessing DNA ploidy in urothelial carcinomas, providing great sensitivity for detecting small aneuploid peaks and multiploid tumors. DNA image analysis of touch preparations is especially useful for carcinoma in situ and small biopsies unsuitable for Hedley's technique. PMID- 9349905 TI - Modeling, definition and applications of histogram features based on DNA values weighed by sine functions. AB - OBJECTIVE: To model new DNA histogram features that weigh DNA values with values of curves of a sine function and to show the definition and applications of such features. STUDY DESIGN: A simple example of a sine feature can be modeled to yield the value zero if all cells are diploid or polyploid, with values of 2c, 4c or 8c, and to yield the value 100 if all cells are aneuploid, with DNA values of 3c, 6c or 12c-e.g., cells that are probably from a malignant lesion or indicate proliferation. All other values are multiplied by the corresponding sine value. We folded the logarithmic DNA histogram with a sine curve with positive values only. RESULTS: Correlation with ploidy balance was -0.94, demonstrating the similarity of both features. The sine features, however, avoid cutpoints between diploid and aneuploid values and are therefore less influenced by minor mistakes in standardization of DNA histograms. We introduced deviation factors as variants; that led to higher sine values for higher c values. For breast carcinoma (N = 306) the sine values were spread from very low to very high values, whereas esophageal carcinomas (N = 125) were centered at a sine value of 50. In breast carcinoma the sine features also correlated with prognostic factors, including hormone receptor status. CONCLUSION: Description of DNA histogram features by graphic demonstration of their weight functions improves understanding of features. Since functions respect the cyclic events in proliferation and are not influenced by polyploidization. PMID- 9349906 TI - Application of the learning vector quantizer to the classification of breast lesions. AB - OBJECTIVE: To investigate the potential of the learning vector quantization (LVQ) neural network for the discrimination of benign from malignant breast lesions. STUDY DESIGN: Using a custom image analysis system on Giemsa-stained smears, 25 parameters describing the size, shape and texture of the cell nucleus were measured. Three thousand nuclei from a total of 9,356 were selected as a training set for the neural network, and the whole data set was used for testing. An additional 238 cells from 16 cases without final cytologic diagnoses were evaluated by the system. The total number of cells (9,594) was collected from 100 patients (68 carcinomas and 32 benign lesions). RESULTS: Cytologic examination of the cases gave two false positive and two false negative results. However, in eight cases of ductal breast carcinoma and in eight cases of benign lesions, histologic confirmation was necessary in order to confirm the cytologic diagnosis. Application of the LVQ permitted correct classification of 87.41% of the cells. Classification at the patient level by using a hypothesis test for proportion with a hypothesis value equal to 50% permitted the correct diagnosis in 98% of patients. CONCLUSION: These results indicate that the use of neural networks combined with image morphometry and statistical techniques may offer useful information about the potential for malignancy, improving the diagnostic accuracy of fine needle aspiration of breast lesions. PMID- 9349907 TI - The politics of identity in epidemiology. AB - The predilection to doubt the objectivity of scientists based on their affiliation--particularly, but not exclusively, focused on scientists affiliated with industry-is a malignant influence in modern epidemiology. Manifestations of this "politics of identity" are widespread and include ad hominem attacks, publication restrictions, and exclusions of scientists from expert panels and advisory boards. These actions are likely to be detrimental to progress in epidemiology and are questionable from an ethical standpoint. The purpose of this commentary is to draw critical attention to the politics of identity in epidemiology and the effects it can have on individual scientists and scientific debate. PMID- 9349908 TI - Equal treatment: bestowed or earned? AB - PURPOSE: While the complaint against non-industry employed epidemiologists for holding their industry-based colleagues to a higher level of scrutiny is accurate, this paper shows that there is a sound basis for such treatment. It also shows, however, that a shift towards ongoing vigilance is needed on the part of all epidemiologists to guard against such bias. METHODS: The proposed shift is made possible through the recent incorporation in ethics guidelines of principles that indeed identify the impropriety of any such bias. RESULTS: In the same guidelines, there are principles that require scientific impartiality. Industry based epidemiologists, by the condition of their employment, may find the avoidance of partiality to the corporate interest more problematic than do non industry based epidemiologists to their respective sponsors. It is in light of past examples of partiality among industry-based epidemiologists that other epidemiologists may be biased against them. CONCLUSIONS: This paper concludes with the realization that both groups of epidemiologists have the challenge of correcting the biases inculcated over many years. Trust needs to be established between industry and non-industry-based epidemiologists through greater acceptance on the part of the latter and exemplary conduct on the part of the former to overcome past practice records. PMID- 9349909 TI - Ibuprofen use in children: a benefit or a risk? PMID- 9349910 TI - Ibuprofen and skin and soft tissue superinfections in children with varicella. AB - PURPOSE: To investigate the possible association between ibuprofen use and dermatologic superinfections among children with recent varicella infection. METHODS: A retrospective cohort study of children in Harvard Pilgrim Health Care, a health maintenance organization in New England, was conducted. Outcomes and exposures of interest were identified from automated medical and pharmacy records. Exposure was defined by dispensing of ibuprofen before varicella to avoid potential confounding by indication. RESULTS: Between July 1, 1990 and September 30, 1994, 89 superinfections developed among 7,013 cases of varicella. The 30-day risk of superinfection was 7.2/10(3) cases (95% CI = 5.8-8.8/10(3) cases). Four of 169 children dispensed ibuprofen within 180 days of varicella developed superinfection. Relative to children without prior use, children with ibuprofen dispensed in the month prior to varicella were 3.1 times more likely to be diagnosed with a superinfection (95% CI = 0.1-19.7; P-value: 0.31). Restriction of outcomes to superinfections treated with systemic antibiotics increased the odds ratio to 5.1 (95% CI = 0.1-32.5; P-value: 0.22). CONCLUSIONS: The results of this study are consistent with a broad range of effects including no association and suggest that further study is needed. PMID- 9349911 TI - Familial aggregation and covariation of diseases in a Japanese rural community: comparison of stomach cancer with other diseases. AB - PURPOSE: We investigated familial aggregation as well as familial covariation of diseases by means of a questionnaire survey dealing with family histories of stomach cancer, stroke, hypertension, diabetes and tuberculosis as well as life style among 2,769 inhabitants of a rural community (84% of census population). METHODS: The strength of familial aggregation was shown by an odds ratio (OR) that compared the number of families in which siblings suffered from one of the above diseases among families in which at least one parent suffered from it, and among families in which neither did. Probands were divided into two groups for analysis: an under-55 "young group," and a 55-and-older "old group." RESULTS: The OR for stomach cancer was lowest and insignificant in the young group, and significant (2.2, p < 0.05) only in the old group. The OR for stroke, hypertension, and tuberculosis was 4.5-5.1 (p < 0.05) in the young group but decreased to 2.3-3.2 in the old group. Diabetes increased from 3.9 to 5.7 (p < 0.05) with advancing age. Age-related OR trends were not affected by exposure to cigarette smoke in the past. Stomach cancer showed a borderline familial covariation with diabetes and a borderline inverse covariation with hypertension. Hypertension showed a familial covariation with stroke and diabetes. CONCLUSIONS: Among the investigated diseases, familial aggregation was weakest for stomach cancer. The results suggest that stomach cancer may share a common familial etiologic factor with diabetes and hypotension. PMID- 9349912 TI - Reproducibility of the HERITAGE Family Study intervention protocol: drift over time. AB - PURPOSE: The primary goal of the HERITAGE Family Study was to document the role of the genotype in the response to aerobic exercise training. Toward this end, nuclear families were enrolled in a 20-week exercise training program, with a large variety of tests performed before and after the training. Since study drift has the potential to adversely affect the results, reproducibility and potential bias over six consecutive 4-month periods were examined for selected test. METHODS: Intraclass correlations (ICC), technical errors (TE), coefficients of variation within subject (CV), and means were calculated with use of the pretraining test results for each of the six time periods. To check for homogeneity, hypothesis tests were performed on the intraclass correlations and means. If homogeneity was not found across all six periods, further tests were performed to assess differences between pairs of time periods. RESULTS: There was little evidence for real drifts in reproducibility, with most tests having ICCs of 0.8 or better. Only a few tests showed any change over time, and in no case was there evidence of a systematic drift in mean values. CONCLUSIONS: Overall, the reproducibility of the HERITAGE Family Study tests and assays considered in this paper was found to be very good, with no evidence of any systematic drift over time. PMID- 9349913 TI - Performance of a shortened telephone-administered version of a quantitative food frequency questionnaire. AB - PURPOSE: We evaluated the performance of a telephone-administered food frequency questionnaire in a study of 190 men and women, 30-79 years of age, who participated as controls in a study of colon cancer. METHODS: The telephone version of the questionnaire was modified from a longer food frequency questionnaire originally administered in person to each of the participants. One month later, the telephone questionnaire was administered to a subgroup of 190 participants and readministered to 169 members of the subgroup two weeks later to assess the reproducibility and comparative validity of the instrument. RESULTS: The unadjusted correlation for energy between the original in-person full food frequency questionnaire and the abbreviated telephone version was 0.69. The median energy intake from the telephone version was 17% lower in men and 23% lower in women. The energy and sex-adjusted correlation coefficients for other nutrients ranged from 0.45 for vitamin E to 0.78 for fiber. The intraclass correlation coefficients to measure reproducibility ranged from 0.62 for animal protein to 0.83 for folate. CONCLUSIONS: These data indicate that this brief, telephone-administered questionnaire is reproducible and provides a ranking of nutrient intake comparable to that provided by a full in-person interview. PMID- 9349914 TI - Evaluating minority recruitment into clinical studies: how good are the data? AB - PURPOSE: There has been much publicized concern about difficulty with minority recruitment into research studies, particularly since minority inclusion in randomized clinical trials was mandated by the 1993 National Institutes of Health Revitalization Act. We reviewed recruitment data in published reports from clinical studies to assess the actual degree of success in recruiting minorities versus whites and to identify barriers to recruitment. METHODS: We abstracted articles published between September 1993 and February 1995 that reported detailed results of participant recruitment for studies conducted in the United States. RESULTS: Of 65 articles meeting our eligibility criteria (median sample size, 1323), only one (1.5%) reported the racial/ethnic composition of potential study participants. Only two articles (3.1%) provided information about the racial/ethnic composition of eligible subjects, and only one (1.5%) provided information about the racial/ethnic composition of refusing subjects. For enrolled subjects, race/ethnicity was less likely to be reported (58.5%) than were age (90.8%) or gender (80.0%). CONCLUSIONS: The published literature currently contains insubstantial data to either refute or prove that there are differential recruitment rates among minorities as compared with whites. Changes in reporting will be needed in order to track progress in this area. PMID- 9349915 TI - Do ethnic differences in dietary cation intake explain ethnic differences in hypertension prevalence? Results from a cross-sectional analysis. AB - PURPOSE: To better understand how the magnitude of the association between ethnicity and hypertension is affected by ethnic differences in dietary cation intake, we describe differences in dietary cation intakes and prevalence of hypertension across four ethnic groups (African-Americans, European-Americans, Mexican-Americans, and Puerto Ricans). We also assess the cross-sectional association between: (i) hypertension and self-reported dietary intakes of sodium, potassium, and calcium for each ethnic group; and (ii) ethnicity and hypertension before and after adjustment for dietary cation intakes. METHODS: Data from the Second National Health and Nutrition Examination Survey (1976-1980) and the Hispanic Health and Nutrition Examination Survey (1982-1984) were analyzed. Multiple logistic regression was used to estimate odds ratio (OR) for hypertension for each ethnic group, with adjustment for age, body mass index (BMI), and diabetes status. Comparisons were made to assess whether the magnitude for the ethnicity ORs changed when the three nutrient variables were entered into the model. RESULTS: Mexican-American and Puerto Rican men and women showed clinically and statistically significantly higher mean intakes of the three cations than did African-American men and women, who reported clinically and statistically significantly lower mean intakes of sodium, potassium, and calcium than did European-American men and women. Mean dietary intakes of potassium and calcium were higher for normotensives than for hypertensives among all ethnic groups, except African-American and Mexican-American women. In multivariate modeling, stark differences in ORs for hypertension persisted across ethnic groups despite inclusion of the nutrient variables. CONCLUSION: In this cross sectional study, adjustment for dietary cation intakes did not alter the magnitude of the ethnic differences in prevalence of hypertension. PMID- 9349916 TI - Blood pressure and heart rate: no evidence for a positive association with prostate cancer. AB - PURPOSE: This study aimed to determine whether blood pressure and heart rate are correlated with the development of prostate cancer. METHODS: A total of 58,704 men whose blood pressure, heart rate, and other characteristics were measured at multiphasic checkups were followed for < or = 30 years. The incidence of, and mortality from, prostate cancer were measured. There were 2297 cases and 464 deaths. RESULTS: No evidence of a positive association of blood pressure or heart rate with subsequent prostate cancer was found. CONCLUSIONS: Data from this large population lend no support for concerns that higher levels of blood pressure or heart rate are linked to increased risk of prostate cancer. PMID- 9349919 TI - Male and female occupation in relation to miscarriage and preterm delivery in central North Carolina. AB - PURPOSE: This study was undertaken to evaluate the role of parental occupation in miscarriage and preterm delivery. Previous studies raise the possibility that both male and female exposures could affect pregnancy. METHODS: Data from a population-based study of miscarriage and preterm delivery in central North Carolina were used to examine potential associations with male and female occupation. Medically treated miscarriage cases (n = 418), preterm delivery cases identified through hospital record review (n = 582), and term, normal birth weight controls (n = 787) were sought for telephone interview. The interview included information on jobs the woman held before and during the pregnancy, reports of her partner's job around the time of pregnancy, and information on potential confounding factors. RESULTS: Female employment overall, or in specific jobs, around the time of conception or early pregnancy was not associated with the risk of miscarriage, whereas working during pregnancy, especially in the seventh month, was inversely associated with risk of preterm delivery. Male employment in several industrial occupations was weakly associated with miscarriage (adjusted odds ratios (OR) of 1.6 to 1.8), and somewhat more strongly associated with preterm delivery, particularly for chemists and sheet metal workers (adjusted OR over 3). Restriction to married men strengthened the associations. CONCLUSIONS: Our results are limited by nonresponse, imprecision, incomplete identification of miscarriages, and lack of detailed occupational exposure information. Nonetheless, we found greater support for further examination of male compared to female jobs in relation to pregnancy outcome. PMID- 9349918 TI - Low-to-moderate gestational alcohol use and intrauterine growth retardation, low birthweight, and preterm delivery. AB - PURPOSE: Heavy drinking during pregnancy is an established risk factor for fetal alcohol syndrome and other adverse perinatal outcomes. However, there is still debate as to the effects of low-to-moderate drinking during pregnancy. METHODS: This prospective investigation was based on 2714 singleton live births at Yale New Haven Hospital during 1988-1992. Alcohol drinking during pregnancy was evaluated with respect to intrauterine growth retardation (IUGR), preterm delivery, and low birthweight. RESULTS: Mild drinking, defined as > 0.10-0.25 oz of absolute alcohol per day, during the first month of pregnancy was associated with a protective effect on IUGR (OR, 0.39; 95% confidence interval (CI), 0.20 0.76). Overall, drinking during month 1 of pregnancy suggested a curvilinear effect on growth retardation, with consumption of > 1.00 oz of absolute alcohol per day showing increased risk. Drinking during month 7 was associated with a uniform increase in the odds of preterm delivery; the ORs were 2.88 (95% CI, 1.64 5.05) for light drinking and 2.96 (95% CI, 1.32-6.67) for mild-to-moderate alcohol consumption. CONCLUSIONS: Differences in the risk estimates for IUGR and preterm delivery may indicate etiological differences that warrant further investigation of these outcomes and critical periods of exposure. Low birthweight is not a useful neonatal outcome for this exposure because it is a heterogeneous mix of preterm delivery and IUGR. Despite the observed protective effects of mild drinking on IUGR, the increased risk of preterm delivery with alcohol use supports a policy of abstinence during pregnancy. PMID- 9349917 TI - Ischemic heart disease and alcohol-related causes of death: a view of the French paradox. AB - PURPOSE: In France the low rates of death due to ischemic heart disease have been attributed to the high consumption of alcohol. However, the question remains: are the higher death rates for causes associated with alcohol consumption an explanation? METHODS: Diseases were defined according to the International Classification of Diseases, revision 9. World Health Organization data on country and age-specific death rates were used. RESULTS: Official causes-of-death statistics for men 40-74 years of age show that in 1990 French men under 50 years old had low death rates from ischemic heart disease but a relatively high all cause mortality rate, in contrast to low rates for men 60 to 74 years of age. Among French men aged 40-44 years in 1960, 34% had died before reaching the age of 70-74 years. In comparison, 37% in the United States and 36% in England and Wales, had died by this age, with 4.5%, 14.1%, and 15.2% of deaths, respectively, due to ischemic heart disease. If all of the men who died early of causes associated with alcohol had died of ischemic heart disease, there would still be a lower rate in France (21%) than in the United States (26%) or in England and Wales (25%). CONCLUSION: Thus, although some of the chronic heavy drinkers in France die early of causes associated with excessive alcohol consumption, this is not the only reason for the low ischemic heart disease death rates. PMID- 9349921 TI - [Hormone induced electrokinetic properties of the isolated nuclei of hepatocytes]. AB - It has been investigated the influence of hydrocortisone and estradiol on conforming objects for revealing the correlation between the electrokinetic potential and the physiological state of nuclear membrane. It has been shown that the induction by different hormones leads to many-direct differences in potential. On the basis of obtained results we conclude that the electrokinetic potential may carry an information about functional condition of nuclear membrane and can be a convenient instrument for indirect determination of its physiological state. PMID- 9349920 TI - The potential for Simpson's paradox in drug utilization studies. AB - PURPOSE: Simpson's paradox is a type of severe confounding wherein a confounding variable changes the direction of an association. METHODS: This article demonstrates Simpson's paradox with three cohorts of naproxen users (new users, chronic users, and combined users) who were compared on the age/sex distribution of further naproxen use. Hypothetical new and chronic user populations were constructed with the same proportions for further naproxen use as their original counterparts. RESULTS: The hypothetical combined population showed an age/sex distribution opposite to that of the original combined population. CONCLUSIONS: This example of Simpson's paradox is a significant finding as many drug utilization studies do not distinguish between component populations. PMID- 9349922 TI - From clinical trials to clinical practice: oral anticoagulation among patients with non-rheumatic, atrial fibrillation. AB - OBJECTIVES: The aim of the present study was to evaluate the impact of the results of clinical trials on the prophylactic treatment of non-rheumatic atrial fibrillation with oral anticoagulants. METHODS: Retrospectively, we studied a random sample of 375 patients discharged from our hospital with a diagnosis of non-rheumatic atrial fibrillation between 1991 and 1993. Information about diagnoses, other clinical variables and treatments prescribed at discharge was obtained from the hospital medical records. RESULTS: During the whole study period, 14% of patients were prescribed an oral anticoagulant agent and 17% were prescribed acetylsalicylic acid. A non-significant increase in the proportion of patients prescribed oral anticoagulant drugs, from 9% to 17%, was observed. Multivariate analysis showed that a history of stroke (OR = 5.96) and younger age were significantly associated with the prescription of oral anticoagulants. ASA prescription was strongly associated with a history of concomitant vascular disease (OR = 5.8), but not with other risk factors for stroke. Sixty-five percent of patients had one or more risk factors for stroke, did not present any contraindications to anticoagulant agents, but nevertheless were not prescribed one of these drugs. CONCLUSIONS: Anticoagulant agents and acetylsalicylic acid were largely underprescribed to patients with non-rheumatic atrial fibrillation, and oral anticoagulants were not prescribed according to the individual patients' risk of stroke. PMID- 9349923 TI - Methods for estimating the occurrence of polypharmacy by means of a prescription database. AB - OBJECTIVE: Concurrent use of multiple drugs (polypharmacy, PP) may cause health risks such as adverse drug reactions, medication errors and poor compliance. The objective of this study, based on data from a prescription database, was to evaluate estimators of PP in the general population. METHODS: Data were retrieved from Odense Pharmaco-epidemiological Database (OPED) and consisted of all prescriptions in 1994 from a 10% random sample of drug users (n = 26,977) in the county of Funen, Denmark. For each prescription, the period of consumption was calculated by setting the duration of treatment to equal the amount of drug purchased, as measured in defined daily doses (DDD), thereby assuming a daily intake of one DDD. PP was defined as overlapping periods of consumption for different drugs. A Venn diagram was used to illustrate and compare this estimator of PP with two other indicators of multiple-drug use: the number of drugs purchased in 3 months and the mean number of drugs used in 1 year. A receiver operating curve (ROC) was used to evaluate the possibility of predicting episodes of PP from the number of drugs purchased in 3 months. RESULTS: The proposed estimator of PP was robust towards changes in DDD. On an average day in 1994, the prevalence of PP was 9.9% and the standard deviation (SD) between days was 0.3%. Two to four drugs (minor PP) were used by 8.7% of the population (SD, 0.2%) and five or more drugs (major PP) by 1.2% (SD, 0.1%). The number of individuals displaying PP for the first time in 1994 stabilised after approximately 6 months, resulting in an incidence of major PP of 0.2% and of minor PP of 1.2% per month. For individuals exposed to PP, the median number of days of exposure was 61 and 10.5% were exposed for more than 350 days of the year. Purchase of five or more drugs in the first 3 months of 1994 predicted episodes of major PP in the same year with a positive predictive value of 80%. CONCLUSION: Epidemiological measures of multiple drug use can be estimated from data in a prescription database. From a conceptual point of view, an estimator based on the number of simultaneously used drugs (calculated from the date of purchase and the number of DDD) is preferable, but the number of drugs purchased in a 3-month period may also be a useful estimator. PMID- 9349924 TI - A medication database--a tool for detecting drug interactions in hospital. AB - OBJECTIVE: Drug interactions may lead to life-threatening injuries. More often, however, they lead to slow recovery, induce slight symptoms or result only in potential injury. Therefore, clinicians are not always aware of using potentially interacting drug combinations. An on-line alarming system of potential drug interactions was developed in Turku University Central Hospital. In the present study, we utilised the system to find out the incidence and nature of potential drug interactions occurring in a representative hospital patient population. METHODS: Computerised anatomical therapeutic chemical (ATC)-coded patient medication data of 2547 patients, treated in two internal medicine wards, were combined with an ATC-coded rule base of drug interactions. All potential drug interactions in the study population were searched for. RESULTS: A total of 326 potentially serious drug interactions were detected in the study population. The number of patients in this group was 173, i.e. 6.8% of all patients had one or several drug combinations which might have led to serious clinical consequences. Concomitant use of calcium and fluoroquinolones (decreased absorption) was the most common mistake (66 prescriptions). CONCLUSIONS: Potentially inappropriate drug combinations seem to occur frequently. Structured and coded medication data can be utilised efficiently to detect potential drug interactions in hospital. Computerised online monitoring and automatic alarming of potentially hazardous drug combinations might help clinicians to prescribe more safely, but further development of the system is needed to avoid unnecessary alarms. PMID- 9349925 TI - Incidence and cost of adverse drug reactions in a French cancer institute. AB - OBJECTIVES: The incidence and the cost of adverse drug reactions (ADR) in patients treated by cancer chemotherapy were assessed using hospital database records from 1993 in a French regional cancer institute. METHODS: Patients with ADRs were identified using a list of ICD-9 codes describing potential adverse events. Direct medical costs for treating these ADRs were assessed according to the hospital system of claims data. RESULTS: Among the 3429 in-patients hospitalized in 1993, we found 171 patients (5% of the population) who presented at least one ADR (3.5% of the total number of hospital stays). A total of 313 ADRs occurred in 256 hospital stays (3.5% of the hospital stays in 1993). Of the patients with ADRs 60.2% were female and their mean age was 51.5 years; 106 patients presented with at least one "serious" ADR according to the WHO definition. These ADRs occurred during 130 hospitalizations. In 7 cases, ADRs led to death. There was no relationship between age or sex and the seriousness of the ADR. Of the ADRs 91% was type "A" (predictable). We estimated that the cost of "serious" ADRs was 1.8% of the global budget of the hospital. The average cost of ADRs leading to hospitalization was 33 037 French Francs at the current rate in 1993. This cost represented an additional cost of 32% of the overall cumulative yearly cost per patient in the institution. CONCLUSION: This study emphasizes the medical and economic impact of the management of ADR in anticancer treatments. PMID- 9349926 TI - Effects and pharmacokinetics of high dose metoprolol on chest pain in patients with suspected or definite acute myocardial infarction. AB - OBJECTIVE: Pain intensity and the plasma concentrations of metoprolol and its major metabolite alpha-hydroxymetoprolol as well as noradrenaline (NA), adrenaline (A) and neuropeptide Y (NPY) were determined in patients with pain due to definite or suspected acute myocardial infarction (AMI) after graded metoprolol infusion. Pain intensity and metoprolol kinetics were assessed over 8 h. METHODS: Twenty-seven patients of either sex, aged 48-84 years with ongoing chest pain upon arrival to the Coronary Care Unit (CCU) were subdivided into two groups: (1) patients with ECG signs of threatening transmural myocardial damage (n = 15); and (2) patients without such ECG signs (n = 12). Pain intensity was assessed by a numerical rating scale (NRS) and venous blood was obtained for determination of plasma catecholamine and NPY concentrations. A continuous infusion of metoprolol (3 mg.min-1 i.v) was started and serial blood samples for plasma catecholamines, NPY as well as metoprolol and its major metabolite alpha hydroxymetoprolol were obtained from the contralateral arm. RESULTS: Initial pain intensity was 5.9 (arbitrary units) and 5.4 in the groups with and without signs of transmural myocardial damage, respectively. One third of the patients with ST changes reported full pain relief (NRS = 0) within 70 min after starting metoprolol infusion (accumulated dose, 15-180 mg). Among the patients without ST changes upon arrival, full pain relief was obtained in 70% (accumulated dose, 30 120 mg). There was a dose-dependent relation between accumulated metoprolol dose and pain relief. The diagnosis of acute myocardial infarction (AMI) was confirmed in all 15 patients with ECG signs on arrival of transmural myocardial damage. The mean metoprolol dose in this group was 91(12) mg. The mean metoprolol dose in the 12 patients without ST changes was 64(8) mg. In all, seven of these patients developed definite AMI. The terminal half-life of unchanged metoprolol ranged from 2.5 to 8.5 h in group 1 and from 2.2 to 5.2 h in group 2. In group 1, metoprolol half-life was 4.5 h and total plasma clearance (CL) 54.1 1.h-1. In group 2, the metoprolol half-life was 3.7 h and total plasma clearance 75.4 1.h 1. There was a significant difference in clearance between the groups. After the intravenous metoprolol infusion, alpha-hydroxymetoprolol concentrations increased gradually. In groups 1 and 2, maximal concentrations in plasma (Cmax) were 143 and 135 nmol.1(-1) for alpha-hydroxymetoprolol and 2830 and 1653 nmol.1(-1) for metoprolol, respectively. Plasma NA or NPY did not differ between the groups. In contrast, plasma A was significantly higher during the initial 90 min of observation in patients with ECG signs of transmural myocardial damage. CONCLUSION: High-dose intravenous metoprolol was well tolerated in patients with suspected AMI. There was a more rapid and almost complete pain relief in patients without signs of transmural ischaemia compared with the patients with ECG signs of transmural AMI at arrival. In the later group of patients, plasma clearance of metoprolol was significantly reduced. PMID- 9349928 TI - A drug interaction study between ticlopidine and cyclosporin in heart transplant recipients. AB - OBJECTIVES: Previous uncontrolled studies have suggested an interaction between ticlopidine, a major antiplatelet agent, and cyclosporin in heart- and kidney transplant recipients. The aims of this study were to examine in a randomised, double-blind fashion, the possible interaction between cyclosporin A and ticlopidine (250 mg per day) and the tolerability of this combination in heart transplant recipients. METHODS: Twenty heart-transplant recipients were randomised into either a treated or a placebo group. Blood samples were drawn for time-course evaluation of cyclosporin blood levels over a period of 12 h, following the morning intake of cyclosporin and, for platelet aggregation studies, before and after 14 days of ticlopidine administration. Twenty four-hour urine samples were collected for 6-beta-hydroxycortisol measurements, before and after 14 days of ticlopidine. RESULTS: Although given at half the recommended daily dosage, ticlopidine significantly reduced platelet aggregation. Pharmacokinetic parameters indicate that the bioavailability of cyclosporin A was not significantly modified by ticlopidine. However, one patient in the ticlopidine group was withdrawn because of a major fall in cyclosporin blood level within 3 days of treatment. Urinary excretion of 6-beta-hydroxycortisol was augmented after treatment in the ticlopidine group compared with the placebo group, suggesting that induction of drug metabolism might have occurred. Data also show quite a large intra-individual variability in cyclosporin bioavailability in the placebo group, suggesting that poor absorption of the drug formulation and/or poor compliance might have contributed to the decreased cyclosporin blood levels in the patient withdrawn from this study and in previous uncontrolled studies. CONCLUSION: Cyclosporin bioavailability was not clearly modified by a half dosage of ticlopidine in this study. We, however, recommend closely monitoring cyclosporin blood levels when prescribing ticlopidine. Further studies will be needed with new formulations of cyclosporin or when using the full dosage of ticlopidine. PMID- 9349927 TI - Adrenal suppression with high doses of inhaled fluticasone propionate and triamcinolone acetonide in healthy volunteers. AB - STUDY OBJECTIVE: This study was conducted to compare the adrenal suppression of inhaled fluticasone propionate and triamcinolone acetonide in healthy volunteers, both given via their respective pressurised metered dose inhaler (pMDI) devices at high doses within the manufacturers recommended dose range. DESIGN: We used a single (investigator) blind, randomised, crossover design comparing a total daily dose of 1.625 mg fluticasone propionate delivered via a pMDI, 1.60 mg daily of triamcinolone acetonide delivered via a pMDI with integrated spacer, or placebo pMDI; each drug was given in two divided doses at 0800 hours and 2200 hours over a 24-h period. Each drug treatment was separated by a 1-week washout. PATIENTS: Twelve normal subjects mean age 27.5 years were studied. MEASUREMENTS: Blood samples were taken for 0800 hours plasma cortisol, i.e. 10 h following the second dose. Ten hour urine collections (2200 hours until 0800 hours) were taken for urinary cortisol and creatinine excretion. RESULTS: For the 0800 hours plasma cortisol (geometric mean, nmol.1(-1) compared with placebo (353) fluticasone propionate (138) produced significant (P < 0.05) suppression (2.57-fold difference), whereas triamcinolone acetonide (263) did not (1.34-fold difference). Fluticasone propionate produced a 1.91-fold greater adrenal suppression than triamcinolone acetonide (95% CI 1.10 to 3.33). Individual subjects with abnormally low 0800 hours cortisol values < 150 nmol.1(-1) (< 5.4 micrograms/dl) were n = 4 for fluticasone propionate and n = 0 for triamcinolone acetonide. Overnight urinary cortisol/creatinine ratio (geometric mean, nmol/mmol) did not show any difference between fluticasone propionate (1.48) and triamcinolone acetonide (1.60), with both producing significant suppression versus placebo (4.01): triamcinolone acetonide 2.50-fold difference (95% CI 1.45 4.24); fluticasone propionate 2.71-fold difference (95% CI 1.57-4.69). CONCLUSION: Fluticasone propionate 1.625 mg/day (pMDI) produced an approximately two-fold greater adrenal suppression of 0800 hours plasma cortisol than triamcinolone acetonide 1.60 mg per day (Oral Inhaler) when given twice daily, and one third of subjects with fluticasone had abnormally low 0800 hours cortisol values < 150 nmol.1(-1) (< 5.4 micrograms.dl-1. There were no differences between the drugs for urinary cortisol excretion. Further dose-ranging studies are required at steady-state in asthmatic subjects in order to see whether differences occur at lower doses on the steep part of the dose-response curve for both plasma and urinary cortisol suppression. PMID- 9349929 TI - Comparative lung delivery of salbutamol given via Turbuhaler and Diskus dry powder inhaler devices. AB - OBJECTIVE: The environmental concerns surrounding the use of chlorofluorocarbons (CFC) have led to a resurgence of interest in dry powder inhaler devices. The aim of our study was to compare two commonly used dry powder inhaler devices, namely the Turbuhaler and Diskus. METHODS: Eight healthy volunteers with a mean (SEM) age of 21 years (0.8) were studied using a randomised single-investigator blind crossover design. Single doses of 1.2 mg salbutamol as Turbuhaler (12 x 100 micrograms) and Diskus (6 x 200 micrograms) were administered over 6 min. Mouth rinsing was performed after every inhalation. Lung delivery from each device was assessed by measuring the early plasma salbutamol profile at 5, 10, 15 and 20 min after inhalation. RESULTS: Significant differences in lung delivery were found between the Diskus and the Turbuhaler for salbutamol Cmax 3.21 vs 4.04 ng.ml-1, respectively and Cav 2.65 vs 3.73 ng.ml-1, respectively. This amounted to a 1.28 fold difference (95% CI 1.09 to 1.45) between these devices for Cmax and a 1.42 fold difference (95% CI 1.57 to 1.66) for Cav. CONCLUSION: We have demonstrated that, in vivo, the Turbuhaler dry powder inhaler produces significantly greater lung delivery of salbutamol than the Diskus. This illustrates that dry powder inhaler devices may have different in vivo deposition characteristics. PMID- 9349930 TI - Pharmacokinetics and tolerability of oral iloprost in thromboangiitis obliterans patients. AB - OBJECTIVE: Iloprost is a potent PGI2 mimetic, which has been shown to be therapeutically effective in several vascular disorders. Due to its rapid clearance from the central compartment, iloprost is administered mainly by i.v. infusion, which limits its use to hospitalized patients. In order to improve pharmacotherapeutic use of this PGI2 mimetic, an oral extended-release (ER) dosage form has been developed, which should mimic plasma level profiles as observed after i.v. infusion and serve as a therapeutic equivalent. METHODS: This trial was performed to investigate the tolerability and pharmacokinetics of iloprost administered perorally, compared with i.v. infusion, in 12 patients suffering from thromboangiitis obliterans (TAO). A dose titration was carried out for 1 week with i.v. iloprost, followed by a p.o. titration and treatment phase of 3 weeks' duration. Pharmacokinetics was investigated at the individually tolerated dose levels; i.e., on days 5-7 (i.v. infusion at 2, 2.5 and 3 ng.kg 1.min-1), and twice during p.o. treatment after b.i.d. administration of 50, 100, 150, 200 or 300 micrograms. RESULTS: Individual tolerability of iloprost varied: 7 patients out of 12 tolerated the maximum i.v. dose of 3 ng.kg-1.min-1; six tolerated the maximum oral dose of 600 micrograms. No patients withdrew from the study due to adverse events. Flush and headache were the most common adverse events and seemed to be related to the study drug. After i.v. infusion of iloprost, dose-normalized (3 ng.kg-1.min-1), steady-state plasma levels were 260 pg.ml-1. Terminal half-life was 0.57 h. Total clearance ranged from 8 to 17 ml.min-1.kg-1. Peroral administration of the ER formulation resulted in dose dependent Cmax and AUC values. AUC values of the first and second daily dose interval, i.e., 0-5 h and 5-11 h after first dosing, were almost identical. Absolute bioavailability was 24%, with the exception of two patients who tolerated only 50 micrograms b.i.d. and exhibited a bioavailability of approx. 60%. The AUC values observed in weeks 2 and 4 were identical, demonstrating low day-to-day variability of iloprost plasma level profiles in TAO patients. CONCLUSION: Based upon pharmacokinetic data, the ER formulation provides an equivalent to the i.v. infusion of iloprost and broadens the range of therapy to nonhospitalized patients. The availability of capsules with 50 and 100 micrograms iloprost enables individual dose titration and pharmacotherapy. Beneficial effects, as observed with i.v. iloprost in TAO patients, should therefore be achievable by peroral pharmacotherapy using the new ER formulation. PMID- 9349931 TI - Population analysis of the non-linear red blood cell partitioning of draflazine following various infusion durations. AB - OBJECTIVE: The pharmacokinetics and non-linear red blood cell partitioning of the nucleoside transport inhibitor draflazine were investigated in 19 healthy male and female subjects (age range 22-55 years) after a 15-min i.v. infusion of 1 mg, immediately followed by infusions of variable rates (0.25, 0.5 and 1 mg.h-1) and variable duration (2-24 h). METHODS: The parameters describing the capacity limited specific binding of draflazine to the nucleoside transporters located on erythrocytes were determined by NONMEM analysis. The red blood cell nucleoside transporter occupancy of draflazine (RBC occupancy) was evaluated as a pharmacodynamic endpoint. RESULTS: The population typical value for the dissociation constant Kd (%CV) was 0.648 (12) ng.ml-1 plasma, expressing the very high affinity of draflazine for the erythrocytes. The typical value of the specific maximal binding capacity Bmax (%CV) was 155 (2) ng.ml-1 RBC. The interindividual variability (%CV) was moderate for Kd (38.9%) and low for Bmax (7.8%). As a consequence, the variability in RBC occupancy of draflazine was relatively low, allowing the justification of only one infusion scheme for all subjects. The specific binding of draflazine to the red blood cells was a source of non-linearity in draflazine pharmacokinetics. Steady-state plasma concentrations of draflazine virtually increased dose-proportionally and steady state was reached at about 18 h after the start of the continuous infusion. The t1/2 beta averaged 11.0-30.5 h and the mean CL from the plasma was 327 to 465 ml.min-1. The disposition of draflazine in whole blood was different from that in plasma. The mean t1/2 beta was 30.2 to 42.2 h and the blood CL averaged 17.4-35.6 ml.min-1. CONCLUSION: Although the pharmacokinetics of draflazine were non linear, the data of the present study demonstrate that draflazine might be administered as a continuous infusion over a longer time period (e.g., 24 h). During a 15-min i.v. infusion of 1 mg, followed by an infusion of 1 mg.h-1, the RBC occupancy of draflazine was 96% or more. As the favored RBC occupancy should be almost complete, this dose regimen could be justified in patients. PMID- 9349932 TI - Effects of fluvastatin therapy on lipids, antioxidants, oxidation of low density lipoproteins and trace metals. AB - OBJECTIVE: Oxidation of low density lipoproteins (LDL) is thought to be an important step in the development of atherosclerosis. Trace metals are involved in oxidative processes. It was of interest to determine whether lipid-lowering therapy with fluvastatin, a potent HMGCoA reductase inhibitor, affected LDL oxidation and trace metal levels. METHODS: Twenty hypercholesterolemic patients were treated with fluvastatin (40 mg twice daily) or placebo for 8 weeks in a double-blind, randomized study. LDL composition, antioxidants and oxidizability as well as plasma zinc, copper, selenium and manganese concentrations were investigated. RESULTS: After fluvastatin treatment, total cholesterol was reduced by 24%, triglycerides fell by 26% and plasma Zn fell by 8%. Cu, Se and Mn changed insignificantly. LDL were separated by ultracentrifugation. LDL were reduced by 18%, LDL-C by 29% and LDL-TG by 19%. The concentrations of alpha-tocopherol and retinol in LDL changed insignificantly. LDL preparations were incubated with copper ions (204 mumol.1(-1) LDL-C/3.2 mumol.1(-1) Cu) and formation of conjugated dienes was monitored at 234 nm for 5 h. Treatment with fluvastatin caused a reduction of diene production by 16% and of diene production rate by 14%, effects being significantly different from placebo (P = 0.02). The change of the lagtime did not reach significance; however, it was positively correlated with the change in LDL alpha-tocopherol. In the placebo group, no significant effects were observed. CONCLUSION: Fluvastatin therapy had lipid-lowering and antioxidative effects. PMID- 9349933 TI - Pharmacokinetics and distribution of 6-mercaptopurine administered intravenously in children with lymphoblastic leukaemia. AB - OBJECTIVE: The pharmacokinetics of 6-mercaptopurine, including cerebrospinal fluid (CSF) distribution, and the erythrocyte 6-thioguanine nucleotide concentrations were determined in children randomised to receive intravenous mercaptopurine for acute lymphoblastic leukaemia (ALL), according to the EORTC protocol ALL n.58881. RESULTS: After 1 month of oral treatment at a dose of 50 mg.m-2.day-1, the pharmacokinetic parameters were determined after the first i.v. administration of 1 g.m-2 (bolus dose of 0.2 g.m-2 followed by an 8-h infusion of 0.8 g.m-2) in 11 patients: systemic clearance was 23.02 1.h-1, volume of distribution was 0.75 l.kg-1, and elimination half-life was 1.64 h. The erythrocyte thioguanine concentrations were measured in the same 11 patients and increased significantly between the beginning and the end of infusion (10 pmol x 10(8) packed RBC) or within 24 h of infusion (223 pmol x 10(8) packed RBC). The CSF concentration was 3.78 mumol.1(-1), 1-6 h after the beginning of infusion (n = 28) and the CSF to plasma ratio was 0.15 (n = 16). In patients receiving the oral dose of 50-165 mg.m-2.day-1 of 6-mercaptopurine, CSF concentrations were below 0.18 mumol.1(-1), 1-24 h after drug intake (n = 67), and the CSF to plasma ratio was not calculated. CONCLUSION: Following the i.v. administration of 6 mercaptopurine, we observed high CSF concentrations of 6-mercaptopurine and an acute increase of erythrocyte thioguanine nucleotide concentrations. The clinical trial (EORTC protocol ALL n[symbol: see text]5881), comparing the oral and i.v. administrations of mercaptopurine, will demonstrate if the i.v. administration reduces the incidence of CNS relapses. PMID- 9349935 TI - Drug-induced attack of bronchial asthma in inpatients: a 20-year survey of the Comprehensive Hospital Drug Monitoring Programme on adverse drug reactions, Berne/St. Gallen. AB - Epidemiological aspects of attacks of bronchial asthma related to drugs are prospectively studied in inpatients of three teaching hospitals in the Comprehensive Hospital Drug Monitoring (CHDM)-programme. Results are based on 34,840 individual patients (among 48,005 consecutive admissions) in the years 1974-1993. Between 1974 and 1993, every patient admitted to any of the three medical clinics in the CHDM programme was monitored for any suspicion of an adverse drug reaction (ADR); every drug exposure period during hospital stay was registered. Nineteen patients (0.05% of the 34,840 individual patients) had at least one attack of bronchial obstruction during hospitalisation, considered as probable or definite ADR. The frequency related to exposure periods in response to penicillins is 0.014%, to non-steroidal anti-inflammatories (NSAIDs) 0.0145, to acetyl salicylic acid (ASA) 0.018%, to paracetamol 0.008% and to beta adrenoceptor blockers 0.26%. Of the 12 patients reacting to a drug with an allergic or idiosyncrasy/intolerance type of bronchial obstruction, 7 had a history of bronchial asthma (extrinsic or intrinsic), and 3 had the diagnosis chronic obstructive pulmonary disease (COPD). A history of bronchial asthma or COPD is confirmed to be a risk factor for this particular ADR. Of the seven patients with a bronchial obstruction to beta-adrenoceptor blockers, five were diagnosed with COPD, while two had neither COPD nor bronchial asthma. The relative risk for this pharmacological reaction in COPD patients was 96 (95% confidence interval 45-208) compared with non-COPD patients in the group of 3244 exposed to beta-adrenoceptor blockers. PMID- 9349934 TI - Investigation into the age-dependence of the pharmacokinetics of cyproterone acetate in healthy male volunteers. AB - OBJECTIVE: To investigate a possible age-dependence of the pharmacokinetics of cyproterone acetate following single oral administration. METHODS: Twenty eight healthy men between 22 and 74 years of age received a single oral dose of 100 mg cyproterone acetate. The pharmacokinetic parameters, area under the serum concentration-time curve, apparent volume of distribution, apparent clearance, terminal half-life and the concentration ratio of 15 beta-hydroxycyproterone acetate/cyproterone acetate were examined for possible age-dependence using regression analysis. RESULTS: The values of area under the serum level-time curve showed high interindividual variability and were not related to age. With regard to apparent clearance and volume of distribution, decreasing and increasing values, respectively, were observed with increasing age. There was also a clear dependence of the terminal half-life on age. Elderly men had values about two times higher (95 h) than men belonging to the younger age groups (45 h). The mean concentration ratio of 15 beta-hydroxycyproterone acetate/cyproterone acetate was 0.8 (0.3) and showed no age-dependent change. CONCLUSIONS: Apparent clearance and apparent volume of distribution of cyproterone acetate showed age-dependent changes. Combined, the two effects cause a clear age-dependence of the terminal half-life of cyproterone acetate. An age-related reduction in liver volume is thought to be mainly responsible for the decrease in hepatic clearance with age. Chronic daily administration of the drug to elderly men may therefore lead to somewhat higher steady-state concentrations in the serum than in young men receiving the same dose. PMID- 9349936 TI - Instrumentation in molecular biomedical diagnostics: an overview. PMID- 9349937 TI - Deoxyribonucleic acids as unique markers in molecular detection. PMID- 9349938 TI - Extraction of DNA from human skeletal remains: practical applications in forensic sciences. AB - We have developed an efficient protocol for the extraction of DNA from bone samples. The protocol precludes decalcification and associates an efficient extraction of the DNA with removal of inhibitors by binding of DNA to silica particles. Two forensic cases are described in detail to demonstrate the usefulness of the method. PMID- 9349939 TI - ICE: a novel and efficient method for isolation of chromosomal ends. AB - The linear chromosomal ends are usually unclonable by general methods due to a lack of restriction sites. We present a novel method, isolation of chromosomal ends (ICE), which has been developed for the efficient isolation of linear DNA ends. PMID- 9349941 TI - Location of regulatory gene in penicillin G acylase operon (pacR) of E. coli D816. AB - Regulatory gene in Penicillin G Acylase operon (pacR) of E. coli D816 was located in a Taq1-Dra1 fragment within the pac structure gene. Two ORFs were found in this fragment and their transcriptional orientations were opposite with pac. ORF2 was determined as pacR by point mutation. PMID- 9349940 TI - Rapid method for detection of point mutations using mismatch binding protein (MutS) and an optical biosensor. AB - This paper describes a new method for detecting DNA point mutations using a mismatch binding protein. The interactions of mismatches and mismatch binding proteins are detected by the optical biosensor technology based on surface plasmon resonance (SPR). PMID- 9349942 TI - Transfer of small YACs to E. coli as large circular plasmids. AB - We have designed a YAC circularization vector, pCIRC3, allowing enrichment of the YAC DNA by exonuclease digestion of the linear yeast chromosomes. Due to the presence of P1 replicon sequences in this vector, the circular YACs would replicate as PACs in Escherischia coli. PMID- 9349943 TI - Kappa-casein alleles in Zebu and cross-bred (1/2 Friesian, 1/4 Jersey, 1/4 Hariana) cattle from India using polymerase chain reaction and sequence-specific oligonucleotide probes (PCR-SSOP). AB - Primers and probes were designed to identify two common alleles CASK-A and CASK B. A local breed of cross-bred cattle produced to improve the quality and quantity of milk, were studied for the alleles of kappa-casein (kappa-casein) gene using polymerase chain reaction and hybridization with sequence specific oligonucleotide probes (PCR-SSPO). PMID- 9349944 TI - Topographical distribution of lactate dehydrogenase activity in human clear eye lenses and in lenses with different types of senile cataract: a histochemical investigation. AB - BACKGROUND: Homogenates of human clear lenses show an age-dependent reduction of enzyme activities. Topographical patterns of enzymes in clear and cataractous lenses can be visualized by histochemistry. MATERIAL AND METHODS: Human lenses were characterized by slit-lamp investigations as bearing different types of senile cataracts. Subsequently, lenses were removed by intracapsular extraction. Clear human lenses served as controls. Bovine lenses served to standardize freeze cutting and incubation for lactate dehydrogenase histochemistry. RESULTS: Bovine lenses show a sharp demarcation between the enzyme reaction of cortical fibers bearing cell nuclei and the non-reacting deeper fibers not exhibiting cell nuclei. Clear human lenses, lenses with deep supranuclear cortical cataracts, and lenses with nuclear cataracts exhibit the same borderline. However, in lenses with a subcapsular cortical cataract only the epithelium and a very thin layer of the most superficially located fibers show positive enzyme reactions. CONCLUSION: In growing clear human and bovine lenses, independent of age, the more peripherally located cortical fibers bearing cell nuclei exhibit strong enzyme histochemical reactions. More centrally located lens areas lacking cell nuclei increase in volume in an age-dependent manner. These lens regions do not exhibit enzyme activities detectable by our histochemical technique. Therefore the lens areas free of histochemical reaction product become larger with increasing age, whereas the peripherally located lens fibers apparently do not change their enzyme activities with age. Thus, homogenates of total lenses show age-dependent reductions of enzyme activities, although enzyme activities remain at a physiological level in cortical lens fibers with recognizable cell nuclei. In lenses with immature supranuclear cortical and (particularly) in lenses with black nuclear cataracts, cortical fibers still can exhibit high enzyme activities. Unexpectedly, also ruptured and broken fibers in immature deep supranuclear cortical cataracts show strong enzyme activities. In contrast, in lenses with (incipient) subcapsular cortical cataracts only the most superficially located lens fibers exhibit some enzyme activity. PMID- 9349945 TI - Autoregulation of human optic nerve head blood flow in response to acute changes in ocular perfusion pressure. AB - BACKGROUND: Studies in animals have demonstrated that optic nerve head (ONH) blood flow (F(onh)) is autoregulated, but there is a lack of evidence for such a process in humans. Therefore, we investigated the relationship between F(onh) and mean ocular perfusion pressure (PPm) in normal volunteers when PPm is decreased through elevation of the intraocular pressure (IOP). METHODS: Laser Doppler flowmetry (LDF) was used to measure relative mean velocity (Velohn), volume (Volonh) and F(onh) of blood at sites of the ONH away from visible vessels, while PPm was decreased in two ways: (1) rapidly, by IOP increments of 15 s duration, and (2) slowly, by IOP increments of 2 min duration, both by scleral suction cup in one eye of each of nine subjects. RESULTS: A rapid and large decrease of PPm of more than 100% induced a decrease of more than 80% in F(onh). With the slower decrease in PPm, F(onh) remained constant down to a PPm of approximately 22 mm Hg (IOP = 40 mm Hg) and then decreased, predominantly due to a decrease in Velohn. Immediately after removal of the suction cup, F(onh) increased transiently by 44% above baseline. CONCLUSIONS: This study demonstrates efficient blood flow autoregulation in the OHN, which is probably brought about by an increase in vascular capacitance. The magnitude of the reactive hyperaemia agrees with the compensatory decrease in ONH vascular resistance during IOP elevation. The time scale of the autoregulatory process and the dependence of the hyperaemia upon duration of IOP elevation suggest a metabolic mechanism of autoregulation. PMID- 9349946 TI - Optic disc morphology in myopic primary open-angle glaucoma. AB - OBJECTIVE: To evaluate the morphology of the optic disc in highly myopic eyes with primary open-angle glaucoma. METHODS: Color stereo optic disc photographs of 44 patients with primary open-angle glaucoma and a myopic refractive error exceeding -8 diopters were morphometrically examined and compared with disc photographs of 571 patients with primary open-angle glaucoma and a myopic refractive error of less than -8 diopters. RESULTS: In the highly myopic group, compared to the control group, the optic disc was significantly (P < 0.0001) larger, the disc shape was significantly (P < 0.0005) more elongated, and the optic cup depth was significantly (P < 0.0001) more shallow. The loss of neuroretinal rim was more concentric, and localized retinal nerve fiber layer defects were found significantly less frequently in the highly myopic group than in the control group. In the highly myopic group, zone beta of parapapillary atrophy was significantly (P < 0.0001) larger. CONCLUSION: The optic disc morphology in primary open-angle glaucoma differs significantly between highly myopic eyes and eyes with hyperopia or low to moderate myopia. The highly myopic eyes are characterized by secondary macro-discs with elongated shape, shallow and concentric disc cupping, large parapapillary atrophy, and low frequency of localized retinal nerve fiber layer defects. Glaucomatous optic nerve damage in highly myopic eyes, compared to eyes with a normal refractive error, is more diffuse than localized. PMID- 9349947 TI - Inverse correlation between endothelin-1-induced peripheral microvascular vasoconstriction and blood pressure in glaucoma patients. AB - BACKGROUND: The potent vasoconstrictor peptide endothelin-1 has been shown to participate in the control of peripheral vascular tone and in the regulation of ocular perfusion. In glaucoma patients vasospasms and arterial hypotension have been identified as risk factors for the progression of glaucomatous damage, and the regulation of endothelin-1 release is disturbed in some of these patients. The aim of this study was to assess the relationship between resting blood pressure and cutaneous vascular responsiveness to endothelin-1 and phenylephrine in patients with glaucoma and in matched controls. METHODS: In 9 patients with primary open-angle glaucoma (POAG), 7 patients with normal tension glaucoma (NTG), and 16 age- and sex-matched controls, endothelin-1 and phenylephrine responses were assessed in the human forearm microcirculation using laser Doppler flowmetry during intra-arterial drug administration. Blood pressure was measured intra-arterially. RESULTS: In contrast to alpha 1-adrenergic effects, endothelin 1 responses were inversely correlated to both systolic (r2 = 0.27, P = 0.05) and diastolic (r2 = 0.54, P = 0.001) blood pressure in glaucoma patients, whereas there was no such correlation in controls. Patients with lower blood pressure values were more sensitive to the vasoconstrictor effects of endothelin-1. Cutaneous responsiveness to endothelin-1 and phenylephrine was similar in glaucoma patients and in controls. CONCLUSION: These results reveal that glaucoma patients appear to have peripheral microvascular abnormalities which are exhibited as altered responsiveness to endothelin-1. Thus, this study supports the hypothesis that endothelin-1-related microvascular dysfunction may be involved in the pathogenesis of glaucomatous damage. PMID- 9349948 TI - Permeability of the blood-retinal barrier in healthy humans. European Concerted Action on Ocular Fluorometry. AB - BACKGROUND: The aim of this study was to compare the inward permeability of the blood-retinal barrier in healthy subjects from six European cities. METHODS: Seventy-two healthy subjects (age 20-70 years) were selected. At 30 min and 60 min after fluorescein injection, fluorescein mass in vitreous was calculated from the concentrations measured along the optical axis of the eye. Non-protein-bound fluorescein (NPBF) concentrations were measured in plasma prepared from blood samples taken 7, 15 and 55 min after injection. Blood-retinal barrier permeability (PBRB) was calculated from the vitreous fluorescein mass and the time integral of NPBF and was corrected for the autofluorescence of ocular tissue and for lenticular light transmittance. RESULTS: Mean PBRB values +/- SD (nm.s-1) were 2.07 +/- 0.54 (Coimbra), 2.01 +/- 0.43 (Frankfurt), 2.24 +/- 0.50 (Ghent), 2.37 +/- 0.56 (Herlev), 1.89 +/- 0.44 (Leiden) and 1.74 +/- 0.38 (Porto). Differences between centers were not significant (P > 0.35). Measurements were reproducible and independent of the time after fluorescein injection (P > 0.50). A PBRB higher than 3.16 nm.s-1 or a value which had increased by 32% was considered abnormal (P < 0.05). CONCLUSION: PBRB values were similar in all centers. The results demonstrate that this is a highly sensitive and reliable method for measuring the permeability of the blood-retinal barrier. PMID- 9349949 TI - Ocular involvement in familial erythrophagocytic lymphohistiocytosis. AB - BACKGROUND: Familial erythrophagocytic lymphohistiocytosis (FEL), a rare, rapidly fatal childhood disorder, is characterized by intermittent fevers, hepatosplenomegaly, cytopenia, hypercytokinemia and lymphohistiocytic infiltration with erythrophagocytosis involving multiple organs. We report the clinical and histological features of two infants with FEL and emphasize the ocular findings. METHODS: Microscopic examination of formalin-fixed, paraffin embedded autopsy material was performed. Immunohistochemical studies were performed in case 1. RESULTS: The first patient presented with clinical and laboratory features and a family history consistent with FEL, and a liver biopsy revealed a lymphohistiocytic infiltrate with erythrophagocytosis consistent with FEL. A deceased brother had been diagnosed with FEL. Autopsy showed widely disseminated lymphohistiocytic infiltrates affecting the liver, spleen, bone marrow, lungs, kidneys and brain. Histologic examination of both eyes disclosed a prominent lymphohistiocytic infiltrate of the optic nerve with destruction of nerve fiber bundles as well as milder infiltrates in the choroid, scleral canals, perineural areas in the orbit and the optic nerve head perivascularly. The second patient also had the typical clinical, laboratory and autopsy findings with similar involvement of most organs, including extensive infiltration of the spleen and bone marrow. Histologic examination of one eye revealed marked lymphohistiocytic infiltration of the entire choroid as well as milder infiltration in the trabecular meshwork, iris, ciliary body, optic nerve, meninges and around the central retinal vein in the optic nerve. CONCLUSION: The findings of this study further define the ocular pathologic features of FEL, which are a part of a generalized, multiorgan disseminated disease. PMID- 9349950 TI - A hemizygous A to CC base change of the CHM gene causing choroideremia associated with pinealoma. AB - BACKGROUND: Although mutations of the CHM gene have been reported in the Caucasian patients with choroideremia, there have been no such reports in non Caucasian patients. We analyzed the CHM gene in a Japanese patient with choroideremia associated with pinealoma. METHODS: The method for screening was a nonradioisotopic modification of single-strand conformation polymorphism (SSCP) analysis. The PCR products from the patient and the carrier were screened and directly sequenced using an automated DNA sequencer. The PCR product of the carrier was also subcloned into a vector and the subcloned products were sequenced. RESULTS: SSCP analysis showed an identical abnormal band shift in the patient and the carrier. Direct sequence analysis showed a hemizygous A to CC mutation at nucleotide 1608 of the CHM gene in the patient, suspected to result in the absence or truncation of the predicted CHM protein. The sequence using both the PCR product and the subcloned DNA of the carrier showed both wild-type and mutant bands indicating a heterozygote. CONCLUSION: The hemizygous mutation was detected in a patient and the heterozygous pattern in his mother, the carrier, suggesting that this mutation caused the disease. PMID- 9349951 TI - Low-dose radiation therapy for age-related macular degeneration. AB - BACKGROUND: The study was carried out to evaluate the effect of low-dose radiation therapy in patients with age-related macular degeneration. METHODS: One hundred eyes of 78 patients (mean age 72 years) with different forms of age related macular degeneration were treated with external beam radiotherapy between 1971 and 1989. In four fractions a total dose of 2 Gy was administered over 7 days. Radiation therapy was performed by the conventional 200-kV technique. The mean duration of follow-up period was 7 years (range 0.5 to 20 years). A control group was composed of 96 eyes from patients with AMD who received no therapy. The mean visual acuity at first presentation and the duration of follow-up was the same as in the treatment group. RESULTS: No difference in visual acuity between the treatment and control groups could be observed. After 1, 2, 5 and 10 years the mean visual acuity was equal in the radiation group and the control group. Even in subgroup analysis regarding only the eyes with exudative forms of AMD, no effect of this treatment strategy could be demonstrated. CONCLUSION: Our results suggest that low-dose radiation therapy in patients with age-related macular degeneration has no beneficial effect. However, it must be considered that the dose of 2 Gy is low in comparison to doses used in recently published studies (5 24 Gy). PMID- 9349952 TI - Intracorneal bovine albumin: an immunologic model of corneal angiogenesis. AB - BACKGROUND: We characterized the neovascularization that follows the intracorneal injection of bovine albumin (BA) in rabbits as a model of corneal angiogenesis. METHODS: New Zealand white rabbits received intracorneal injections of phosphate buffered saline with and without various amounts of BA. The rabbits were co sensitized or pre-sensitized by intramuscular BA or were not sensitized. The corneal vascular response was quantified by ranking photographs taken periodically after the injection. RESULTS: In pre-sensitized animals, blood vessels were apparent within 4 days and reached maximum intensity 14 days after the intracorneal injection. Corneas also vascularized in non-sensitized rabbits, but a larger dose (> 0.2 mg BA) was required than in pre-sensitized animals (> 0.02 mg BA). Vascularization began later in non-sensitized animals and was less extensive than in pre-sensitized animals. CONCLUSION: The intracorneal injection of BA is a reproducible model of corneal angiogenesis in rabbits and should allow the involved immunological mechanisms to be elucidated. PMID- 9349953 TI - Proliferative response of cultured human tenon's capsule fibroblasts to platelet derived growth factor isoforms. AB - BACKGROUND: Although platelet-derived growth factor (PDGF) has been thought to be critical in the wound-healing response of Tenon's capsule fibroblasts after glaucoma filtration surgery, no information is currently available concerning the proliferative effect of PDGF isoforms on this cell type. The aim of the present study was to evaluate the proliferative effect of PDGF-AB heterodimer and PDGF-AA and -BB homodimers on cultured human Tenon's capsule fibroblasts. METHODS: Human Tenon's capsule fibroblasts, cultured under serum-free conditions, were stimulated with PDGF-AA, -AB and -BB isoforms in concentrations ranging from 1 to 100 ng/ml. Cell numbers were determined on days 1, 3, 5 and 7, using a cell counter. RESULTS: Addition of PDGF-AB and -BB led to a dose-dependent increase in cell proliferation. A maximal response (79.9% over control) was obtained after 7 days with 30 ng/ml of PDGF-BB, with an EC50 of 8.9 ng/ml. The maximal increase in cell proliferation caused by PDGF-AB (30 ng/ml) was 54.9%, with an EC50 of 12.5 ng/ml. Stimulation with PDGF-AA revealed a significant effect only with concentrations higher than 30 ng/ml. CONCLUSION: Our results indicate that PDGF AB and -BB isoforms are potent stimulators of proliferation of human Tenon's capsule fibroblasts, suggesting that PDGF-AB and -BB isoforms play an important role in the wound-healing response after glaucoma filtration surgery. PMID- 9349954 TI - Primary vitrectomy without scleral buckling for rhegmatogenous retinal detachment. PMID- 9349955 TI - Osteoarthrosis after the Max Lange procedure for unstable shoulders. AB - Thirty-four patients were followed for a mean of 14.4 years after the Max Lange operation for instability of the shoulder. This procedure consists of an anterior capsulorraphy with lateralisation of the subscapularis tendon and the insertion of a bone block at the inferior and anterior part of the glenoid. The Rowe score showed 87% good and excellent results. External rotation and abduction were significantly limited as compared to the normal side. Radiographs revealed osteoarthrosis in 47%. Double contrast computed tomography in 18 cases showed a bony defect in the anterior and inferior part of the humeral head in 12 who had significantly limited external rotation as compared to those without the defect. Lateralisation of the subscapularis tendon with limitation of external rotation may lead to increased contact stress and shear forces at the anterior rim of the glenoid which contributes to the development of osteoarthrosis. PMID- 9349956 TI - The use of a resorbable augmentation device to secure plating of osteoporotic bones. An in vitro study. AB - Osteotomies in porotic human cadaveric humeri were fixed with metallic plates and screws in 3 experimental groups. In group I, osteotomies of the 3 arbitrarily chosen humeri were plated without reinforcement. The osteotomies of the contralateral 3 humeri were plated after reinforcement with a poly(L-lactide) augmentation device. In group II, osteotomies on one side were plated without reinforcement, but those of the contralateral humeri were plated after reinforcement with methylmethacrylate. In group III, the osteotomies of 3 humeri were plated and reinforced with poly(L-lactide) and in the 3 contralateral humeri were plated after reinforcement with methylmethacrylate. The pullout strength for screws from bones augmented with poly(L-lactide) was identical to those where methylmethacrylate had been used. The bones with poly(L-lactide) augmentation had a higher torsional stiffness than those with methylmethacrylate cement and those which were not reinforced. Resorbable polymeric medullary augmentation devices can be used to enhance plating of osteoporotic bones. PMID- 9349957 TI - A prospective study of the accuracy of clinical examination evaluated by arthroscopy of the knee. AB - The diagnostic accuracy of clinical examination for internal derangement of the knee were evaluated by arthroscopy in 195 patients (200 knees). Radiographs were available and 50 patients had magnetic resonance imaging. The clinical diagnosis was correct in 104 knees (52%), incomplete in 70 (35%) and incorrect in 26 (13%). When there was a single abnormality the diagnosis was correct in 70%, but when more than 3 lesions were discovered the figure was 28%. All individual lesions were diagnosed correctly in more than 90%. The lesions most difficult to diagnose were chondral fractures, partial tears of the anterior cruciate ligament and loose bodies. Knees with acute lesions and those with a single diagnosis were significantly easier to diagnose (p < 0.01). Age, sex, MRI and the surgeon were not significant. PMID- 9349958 TI - Idiopathic genu valgum treated by epiphyseodesis in adolescence. AB - Twenty-three patients (13 boys and 10 girls) with the adolescent type of genu valgum were treated by temporary medial epiphyseodesis of both distal femurs with Blount staples (46 knees); 5 also underwent stapling of the proximal tibia. Evaluation before the operation and during the follow up was based on growth charts, photographs and measurement of the intermalleolar distance. The indication for operation was increasing genu valgum with 10 cm or more of intermalleolar separation; the median distance before operation was 13 cm for boys and 12 cm for girls. The staples were removed after a median time of 11 months. At skeletal maturity the median intermalleolar separation was 3 cm for boys and 2 cm for girls. The median frontal angle before operation was 14 degrees for boys and 6 degrees at follow up, the corresponding figures for girls being 14 degrees and 4 degrees. In 2 cases the deformity recurred because the staples were removed too early. In another case a bony bridge formed after the staple was removed and a varus deformity developed; the bridge was resected and the knees became valgus again, but 1 cm of shortening remained on one side. There were no other complications. PMID- 9349959 TI - Medial meniscus replacement by a fat pad autograft. An experimental study in sheep. AB - The medial meniscus in 15 sheep was replaced by a pediculated infrapatellar fat pad graft and resulted in the development of a macroscopically meniscus-like structure within 6 months. Five additional sheep with a meniscectomy were controls. Degenerative changes in the fat pad autograft were visible after one year. Osteoarthritis of the weightbearing medial compartment was detected after 6 months. A temporary protective effect on the cartilage could be attributed to the autograft, but the long term results indicated that this was not permanent. Fat is not suitable as a meniscal substitute. PMID- 9349960 TI - Collagen fibres and mechanoreceptors in regenerated menisci of rabbits. AB - Total medial meniscectomy was carried out on 30 mature rabbits; subsequent regeneration of the meniscus and degenerative joint changes were observed. Out of 30 knees, 22 had regenerated menisci and 10 of these had not developed gross degenerative changes after 6 months. Collagen fibres were arranged irregularly in the regenerated menisci, although in some areas they occurred in parallel. Every type of mechanoreceptor was found in the outer third of the normal menisci, and in the middle and outer third of the regenerated menisci with no statistical difference in numbers. Mechanoreceptors can regenerate in regenerated menisci and may protect the join from degeneration. PMID- 9349961 TI - Subacute osteomyelitis presenting as a bone tumour. A review of 21 cases. AB - Twenty-one patients with subacute osteomyelitis who were initially considered to have bone tumours were reviewed, with an average follow up of 3 years. The clinical symptoms were not specific and laboratory investigations were normal. The radiographic findings were limited osteolysis surrounded by bone sclerosis in 14 cases, osteolysis without definite borders in 6, and onion-layer periosteal bone formation in one. The preoperative diagnoses included osteoid osteoma, osteosarcoma, chondroblastoma, Ewing's sarcoma, giant cell tumour, fibrosarcoma, eosinophilic granuloma, and bone tumour of unknown aetiology. The definitive diagnosis was made by surgical biopsy, histology and cultures which grew staphylococcus in 9 cases. The gross specimens all showed lymphocytes, plasma cells and granulation tissue with osteogenesis. All the patients recovered completely; 17 were treated with antibiotics and immobilisation, and 4 did not need an antibiotic. There was no recurrence of infection after curettage and excision of the infected tissues. PMID- 9349962 TI - Limb salvage in advanced chronic osteomyelitis in children. AB - Chronic osteomyelitis of the entire tibial shaft was treated in 9 children by medical and surgical management in 3 stages. In stage 1, the involved shaft was excised with debridement of the periosteum. Stage 2 consisted of continuous suction-irrigation of the periosteal space with local and systemic antibiotics. In stage 3, the ipsilateral fibular shaft was transferred into the tibial periosteal envelope with the muscles of the middle third and its blood supply. After 18 months, the transferred fibula grew into a new tibia in every case so that the children were able to carry out all their activities of daily living. This method remains the best way of salvaging the limb when the entire tibia is involved. PMID- 9349963 TI - Cardiopulmonary and haemodynamic changes during total hip arthroplasty. AB - We report a prospective study of 30 patients who underwent total hip arthroplasty for osteoarthrosis in order to investigate the haemodynamic and respiratory changes which occur during operation. Cement was used in 17 cases and the implants were not cemented in 13. Pulmonary and cardiac function, blood levels of methylmethacrylate monomer, intramedullary pressure and transoesophageal echocardiography were recorded. Two well differentiated echogenic patterns appeared consistently during the operation. The intramedullary pressure became raised as the cement was inserted. The following changes occurred within seconds and continued for some minutes: elevation of mean arterial pressure and mean pulmonary artery pressure; decrease of arterial oxygen tension and of mixed venous PO2, and greater tissue consumption of oxygen. Although we recorded raised concentration of methylmethacrylate monomer in venous blood after the cement was inserted, there is no evidence that the monomer is responsible for the haemodynamic changes which take place. PMID- 9349964 TI - Sequential mechanical and pharmacological thromboprophylaxis in the surgery of hip fractures. A pilot study. AB - Two hundred and thirty-eight patients with femoral neck fractures were entered into a randomised pilot study comparing the use of sequential treatment by 'Flowtron DVT' garments in the perioperative period followed by Enoxaparin (Clexane-Rhone-Poulenc Rorer), and Enoxaparin alone. One hundred and ninety-three patients were excluded indicating the difficulty of achieving pure comparisons in this population. The remaining 44 were randomised: 21 received Enoxaparin from the time of admission, and 23 had sequential treatment. There was no statistically significant difference in the incidence of thromboembolism. Patient preference did not indicate a favoured treatment subjectively. The operation field was drier in the sequential group, although this did not reach significance. Sequential treatment was not shown to be better or worse than treatment with Enoxaparin, but the trends favoured sequential treatment rather than drug treatment alone. The technique allows the operation to be carried out without the problems produced by low dose heparins and mobilisation is not hindered by compression garments. PMID- 9349965 TI - The use of a hip score in assessing the results of treatment of proximal femoral fractures. AB - An assessment of Salvati and Wilson's hip function rating system was performed after treatment in 207 proximal femoral fractures. The system had little correlation with pain and very poor correlation with change in mobility since injury. The use of composite scores should be abandoned in these circumstances. PMID- 9349966 TI - An extended iliofemoral approach for total arthroplasty in late congenital dislocation of the hip: a case report. PMID- 9349967 TI - The reaction to nailing or cementing of the femur in rats. A microangiographic and fluorescence study. AB - Bone reaction to cement and to a cementless stem was studied in the rat femur with histological fluorescence and microangiographic techniques. Periosteal and endosteal apposition, and consequent remodelling, appeared as a reaction to reaming rather than caused by cement or a cementless stem. Every change in bone began with proliferation, progression and orientation of the vessels. Endosteal apposition was absent in cemented femurs because the entire medulla was occupied by the acrylic cement, but remodelling of the subendosteal cortex followed medullary revascularisation which was far advanced after 90 days. In cementless stems, endosteal apposition of primary woven bone and remodelling was the basis for bony ingrowth and anchorage through bony bridges. Our results suggest that the pattern of blood supply is relevant to the structural organisation of mature lamellar bone around the implant. Cemented stems have maximum anchorage and stability as soon as they are inserted, but this decreases with time as revascularisation occurs. Cementless stems can reach maximum integration later after insertion, and revascularisation is less critical because they usually do not fill the canal completely. PMID- 9349968 TI - Lengthening of an amputation stump by the Ilizarov technique. A case report. AB - A woman, 29 years of age, sustained a below knee amputation following injury and was left with a stump 5.5 cm long. Good soft tissue cover was obtained with a myocutaneous flap using the gastrocnemius muscle and heel pad flap from the injured leg. The stump was then lengthened by 7 cm, to 12.5 cm, using the Ilizarov distraction technique. The patient was able to bear weight on the end of the fixator during the 8 months it was in position. The gap filled with bone and there were no complications. She recovered good function. PMID- 9349970 TI - Multiple rib fractures associated with severe coughing--a case report. PMID- 9349969 TI - Sepsis due to group G Streptococcus after a total hip arthroplasty. A case report. AB - A patient with a total hip arthroplasty developed an aggressive infection with group G Streptococcus. Very few similar cases have been reported, but they all resolved with antibiotics or drainage. A Girdlestone resection was necessary in our case because of loosening and extensive bony destruction. The true incidence may be greater than that reported and the prognosis may be worse. PMID- 9349971 TI - Family functioning 3 years after infantile colic. AB - Infantile colic causes stress to many families during the first weeks of an infant's life. In our previous studies, we found that families with severely colicky infants had more problems in their daily functioning than did families without colicky infants and that the affective state in these families was anxious and conflicted. These characteristics showed some stability from the colicky period to 1 year of age. In the present study, we examined the functioning of these families 3 years after the colicky period. The McMaster Family Assessment Device was used to evaluate the family interaction in 59 families with previously colicky infants and 58 control families. Three years after the colicky period, families with moderately and severely colicky infants did not differ significantly from control families with respect to psychological family characteristics. PMID- 9349972 TI - Concordance of maternal and teacher ratings of school and behavior problems in children of varying birth weights. AB - To examine the concordance of mother and teacher ratings of children born at different birth weights on measures of school functioning, behavioral problems, and social competencies, we used a prospective cohort study involving children in two previously studied multisite birth cohorts whom we recontacted at 8 to 10 years of age. This provided a multisite sample of 784 low birth weight children and 334 normal birth weight children. Teacher reports of children's behaviors were obtained from 80% of the 1400 teachers contacted. We found that birth weight and neonatal health were associated with both maternal and teacher reports; that maternal characteristics, e.g., low levels of education and poor mental health, were associated with the greatest discrepancies in reports; and that although mothers' reports of objective measures were accurate, their assessments of behavioral problems and social competence often differed from those of teachers. PMID- 9349973 TI - Prediction of preschool behavioral problems in healthy and pediatric samples. AB - In a prospective study of 137 children (47 with cystic fibrosis, 48 with congenital heart disease, 42 with no chronic illness), four domains were examined as predictors of parent-reported behavioral problems, particularly internalizing problems, at 4 years of age: child health, child temperament, parent-child relationships, and family environment. Family environment, as measured by the Parenting Stress Index at 1,2, and 3 years, was the most powerful predictor. This suggests that this index is useful as an early screen for children at risk for behavioral problems and that a reduction of parenting stress is an appropriate target of preventive interventions. PMID- 9349974 TI - Child temperament, parenting discipline style, and daytime behavior in childhood sleep disorders. AB - Fifty-two children without significant sleep disturbance seen at a primary care clinic for well-child care were compared on measures of temperament, parenting style, daytime behavior, and overall sleep disturbance to three diagnostic subgroups identified in a pediatric sleep clinic: children with obstructive sleep apnea (n = 33), parasomnias (night terrors, sleepwalking, etc.) (n = 16), and behavioral sleep disorders (limit-setting disorder, etc.) (n = 31). The mean age of the entire sample was 5.7 years. Temperamental emotionality in the behavioral sleep disorders group was associated with a higher level of sleep disturbance (p < .001); parenting laxness was associated with sleep disturbance in the general pediatric population (p < .01); and intense and negative temperament characteristics seemed to be associated with clinically significant behavioral sleep disturbances. Ineffective parenting styles and daytime disruptive behaviors were more likely to be associated with the milder sleep disturbances found in children in a primary care setting. PMID- 9349975 TI - Development of a supplement to the HOME Scale for children living in impoverished urban environments. AB - A Supplement to the HOME (Home Observation for Measurement of the Environment Scale) for impoverished Families (SHIF) was developed for use with young children living in impoverished urban environments. After interviews with clinicians and pilot studies with families, we developed 20 items and added them to the HOME. The supplement was field tested in a sample of 73 high-risk families to evaluate its psychometric properties and ease of use. During the home visit, the Nursing Child Assessment Feeding Scale and the Nursing Child Assessment Teaching Scale were also administered to examine construct validity. Results indicated that the SHIF provided new clinical data, was easy to administer, and, when added to the HOME, had good psychometric properties, e.g., high inter-rater reliability, internal consistency, item-total reliability, and intact construct validity. The SHIF offers a reliable and valid addition to the HOME for use with young children living in impoverished urban environments. PMID- 9349977 TI - The use of family drawings by children in pediatric practice. PMID- 9349976 TI - Anorexia nervosa in a 7-year-old girl. AB - We report on a 7-year-old girl with anorexia nervosa and consider factors contributing to this early emergency. Cognitive differences in younger children can alter their understanding of this illness, so we chronicled this girl's treatment because it diverged from practices used with older patients. Accordingly, effective interventions in very young anorexics might require modifications of treatments used in postpubertal populations. PMID- 9349978 TI - Conceptual issues in developmental screening and assessment. AB - Effective screening requires an understanding of underlying conceptual issues and their relationship to pragmatic concerns. Pragmatic concerns include the concepts that there are many underlying reasons for an "abnormal" screening result; that sensitivity and specificity should be combined with relative risk when considering developmental outcome; and that patterns of congruence among motor, language, cognitive, and adaptive/personal social areas of development should be considered. Important conceptual issues include the following: there is continuity of underlying processes or functions in development; canalized behaviors might give the appearance of discontinuity; integrated functions are more predictive of later developmental levels than are individual functions; the "window" of assessment and the developmental emergence of a specific function will affect screening results; one must consider biologic and environmental risks and their specific effects; and different types of neural structures and their relationship to environmental input help to explain why screening results vary over time. PMID- 9349979 TI - A 6-year-old boy who is displaying "gender-atypical" play behavior. PMID- 9349980 TI - Effects of epidermal growth factor and insulin on migration and proliferation of primary cultured rabbit gastric epithelial cells. AB - We assessed the influence of epidermal growth factor (EGF) and insulin on gastric epithelial restoration in vitro. Rabbit gastric epithelial cells were cultured and formed a complete monolayer cell sheet in 2 days. We created a wound (1.8 +/- 0.05 mm2) by denuding an area of cells, and EGF (0.1-30 ng/ml) and/or insulin (1 nM-1 microM) was added. The restoration process, which included cell migration and proliferation, was monitored by measuring the cell-free area every 12 h for 2 days. Proliferating cells were detected by sequential staining with bromodeoxyuridine (BrdU). Control cells showed complete repair in 36-48 h and restoration was accelerated dose-dependently by EGF or insulin. EGF plus insulin further accelerated restoration, which was then completed in 12-24 h. EGF and/or insulin increased the number of BrdU- positive cells. The results indicated that EGF and insulin additively accelerated gastric epithelial wound repair by stimulating both the migration and the proliferation of gastric epithelial cells (particularly the former). PMID- 9349981 TI - Role of acetylcholine and gastrin-releasing peptide (GRP) in gastrin secretion. AB - Using an isolated rat stomach infusion model, we investigated the role of gastrin releasing peptide (GRP) and acetylcholine in the secretion of gastrin (which plays a major role in gastric acid secretion), and the relationship between gastrin secretion and stomach pH. Bombesin, which has a structure analogous to that of GRP, was used in the experiment. We also investigated whether acetylcholine has muscarine-like or nicotine-like action. Our findings pointed to the presence of an alternative, GRP-mediated, route for stimulating gastric secretion from G cells, other than the acetylcholine-mediated route. We injected bombesin to confirm the presence of such a GRP-mediated route; significantly increased gastrin secretion was observed, even under acidic conditions, in the gastric lumen, which has been considered to show almost no gastric secretion. This secretion was not inhibited by atropine. The results suggested that there are two routes for inducing gastrin secretion from G cells: an acetylcholine mediated route and a GRP-mediated route (intramural peptide neurons). As GRP induced gastrin secretion, regardless of stomach pH, GRP was considered to be more closely related to gastrin secretion. The results also suggested that a muscarine-like action, particularly in the M3 receptor-mediated route, plays a significant role in acetylcholine-mediated gastrin secretion and that nicotine like action is not involved in gastrin secretion. PMID- 9349982 TI - Endoscopic follow-up study of development of gastric antral vascular ectasia associated with liver cirrhosis. AB - Gastric antral vascular ectasia is an important cause of chronic gastrointestinal blood loss. However, its development and progression have not yet been clarified. We investigated its early lesions and progression by reviewing endoscopic films of five patients with gastric antral vascular ectasia followed for liver cirrhosis. In all patients, early findings were prepyloric red spots. In two patients, anemia due to gastrointestinal bleeding was already observed when vascular lesions were confined to the distal antrum. In the other three patients, anemia was observed 1-2 years after they showed a diagnostic pattern of gastric antral vascular ectasia. The vascular lesions gradually thickened and extended throughout the antrum, with the complete picture shown in 1.5-5 years. The pattern of distribution was classified into three types: diffuse spotty, diffuse confluent, and striped. These types could be predicted before the complete formation. Gastric antral vascular ectasia associated with liver cirrhosis started as prepyloric red spots and extended to the proximal antrum in various ways and varying time courses of less than 5 years; this entity may cause hemorrhage even in the early stage. PMID- 9349983 TI - Effect of ecabet sodium on Helicobacter pylori adhesion to gastric epithelial cells. AB - Helicobacter pylori is a major etiological agent in gastroduodenal disorders, with the adhesion of H. pylori to gastric epithelial cells being the initial step of H. pylori infection. Inhibition of H. pylori adhesion is thus a therapeutic target in preventing H. pylori infection. We evaluated the effect of ecabet sodium, an antiulcer agent, on H. pylori adhesion to gastric epithelial cells, using our previously established enzyme-linked immunosorbent assay. The adhesion of H. pylori was significantly inhibited by ecabet sodium in a dose-dependent manner. Our studies suggest that ecabet sodium inhibits the adhesion of H. pylori to gastric epithelial cells. PMID- 9349984 TI - Intestinal Behcet's disease--pathognomonic changes in intramucosal lymphoid tissues and effect of a "rest cure" on intestinal lesions. AB - To clarify the pathognomonic changes of intestinal lesions of Behcet's disease and to determine effective therapeutic measures, we recruited 13 patients with the intestinal form of this disease for study. We performed pathology studies on the resected specimens of 7 patients and treated 5 of the other 6 patients with a low-residue diet. Pathology examination revealed that 6 of 7 had inflammatory ulcerations in the ileocecal region. The ileal ulcers were mainly on the antimesenteric side. We observed remnants of Peyer's patches at the margins of the major ulcerative lesions in 2 of 2 patients examined. There were aggregations of lymphocytes resembling destroyed lymph follicles in the superficial layer at the mouths of small fissuring lesions, and ulcer scars were also noted in Peyer's patches in 4 of 5 other patients. X-ray and endoscopic examinations revealed the disappearance of intestinal lesions in 5 patients within 1 month during, or following the low-residue diet treatment. We found the intestinal lesions of Behcet's disease at sites coinciding with intramucosal lymphoid tissue. The "rest cure" for the affected bowel was effective, i.e., there was significant alleviation of gastrointestinal symptoms and the intestinal lesions disappeared. We speculated that acute exudative inflammation, abscess formation, and consequent ulceration may occur in these tissues by the same mechanisms as those that operate in the positive needle-prick reactions seen in patients with Behcet's disease. PMID- 9349985 TI - Cyclosporine A inhibits interleukin-8 production in a human colon epithelial cell line (HT-29). AB - Intestinal epithelial cells produce various inflammatory mediators. However, the way in which immunosuppressive agents influence the production of these mediators by intestinal epithelial cells is not understood. The effects of cyclosporine A (CsA), tacrolimus (FK506), and dexamethasone (DEX) on cytokine-induced production of interleukin (IL)-8 in a human colonic cancer cell line (HT-29) were examined. HT-29 cells were stimulated with either IL-1 beta or tumor necrosis factor alpha (TNF alpha) together with CsA, FK506, or DEX. The presence of IL-8 protein was detected by enzyme-linked immunosorbent assay, and the expression of IL-8 messenger RNA (mRNA) by reversetranscription polymerase chain reaction. CsA (1, 5, and 10ng/ml) significantly reduced IL-1 beta-induced IL-8 production (by 32%, 41%, and 48%, respectively), and reduced TNF alpha-induced IL-8 production (by 21%, 42%, and 50%, respectively). FK506 or DEX had no effect on IL-1 beta- or TNF alpha-induced IL-8 production. The expression of IL-8 mRNA was also inhibited by CsA. These findings suggest that CsA may influence the production of inflammatory mediators in colonic cells in a different manner from FK506 and DEX. PMID- 9349986 TI - Association between frequency of amino acid changes in core region of hepatitis B virus (HBV) and the presence of precore mutation in Japanese HBV carriers. AB - To elucidate the relationship between the frequency of core mutations and precore mutation of hepatitis B virus (HBV) in Japanese HBV carriers, we investigated the nucleotide sequence of the precore/core region of HBV in 26 Japanese HBV carriers [15 who were HBe antigen-negative (HBeAg-) and 11 who were HBeAg-positive (HBeAg+)]. The number of amino acid changes (5.9 +/- 3.8) in the core region of HBV in HBeAg-carriers was significantly greater than that in the HBeAg+ carriers (1.5 +/- 1.0; P < 0.005). The precore stop codon mutation was found in 93.3% of HBeAg-negative HBV carriers, while no precore mutation was found in the HBeAg positive HBV carriers, suggesting that the frequency of core mutations may be associated with the presence of the precore stop codon mutation. However, there was no significant difference in the frequency of amino acid changes among HBeAg HBV carriers. The mean number of core amino acid changes of liver cirrhosis patients, chronic active hepatitis patients, chronic persistent hepatitis patients, and asymptomatic carriers were 2.7 +/- 1.5, 6.0 +/- 2.2, 4.7 +/- 1.2, and 8.4 +/- 5.3, respectively. We detected hot spots for core mutations, which showed characteristic localizations and specific substitutions: Gly-87, Leu-97, and Thr-130 were detected exclusively in patients with chronic liver disease with or without HBeAg. To address further the relationship between frequency of core mutations and the presence of the precore stop codon mutation, we investigated the precore/core nucleotide sequence serially along with seroconversion in three patients with chronic hepatitis B in whom the hepatitis either became inactive or remained active after the seroconversion. Emergence of the precore stop codon mutation and a significant increase in core amino-acid changes after seroconversion were noted in all three patients. Our results suggest a close association between the frequency of core amino acid changes and the presence of the precore stop codon mutation; some characteristic core mutations may be associated with the clinical course of chronic hepatitis B in Japanese patients. PMID- 9349987 TI - Production and secretion of pancreatic secretory trypsin inhibitor in normal human small intestine. AB - The aim of this study was to prove the production and secretion of pancreatic secretory trypsin inhibitor (PSTI) in human small intestine. To achieve this we analyzed the content of immunoreactive PSTI (irPSTI) in rinsing fluid from isolated small intestine, using the urea method to estimate the volume of epithelial lining fluid recovered. IrPSTI, measured by an enzyme-linked, immunosorbent assay (ELISA), was present in both free and complexed form. The free PSTI showed intact biologic activity, binding trypsin in stable complexes. The complexed PSTI was dissociated on acidification. With the reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blot hybridization, PSTI mRNA was demonstrated in the mucosa of the ileum. These findings indicate that PSTI is produced and secreted in the small intestinal epithelium and may be part of defence system in intestinal mucosa. PMID- 9349988 TI - Gallbladder emptying and cholecystokinin and pancreatic polypeptide responses to a liquid meal in patients with diabetes mellitus. AB - To investigate gallbladder motility and its regulation in patients with diabetes mellitus (DM), we examined the gallbladder response to an intraduodenal test meal by measuring the temporal course of plasma cholecystokinin (CCK) and pancreatic polypeptide (PP) levels. Eighteen patients with type 2 DM and 7 healthy subjects (controls) were enrolled. The gallbladder volume was calculated by the sum-of cylinders method from ultrasonographic images, and plasma CCK and PP were measured by radioimmunoassays. No significant difference was found in either the gallbladder response or in the CCK response between patients with DM and controls. However, the fasting plasma PP level of patients with DM was more than tenfold higher than that of controls. The integrated PP response (IPPR) of patients with DM to the test meal was 8.3-fold higher than that of controls. When patients with DM were grouped according to whether they had been treated with insulin or not, the fasting plasma PP of patients with DM without insulin treatment was significantly higher than the level in those treated with insulin. These results suggest that overproduction of PP-like immunoreactive substance(s) may occur in patients with DM, but the high plasma PP immunoreactivity does not appear to be related to the fasting gallbladder volume or to gallbladder emptying and filling. PMID- 9349989 TI - Macroscopic and stereomicroscopic diagnosis of superficial flat-type early carcinomas of the gallbladder. AB - Superficial flat early carcinomas of the gallbladder are rarely detected clinically. We previously reported that these carcinomas display granular, flat, or gastric area-like surface mucosal patterns. However, these patterns are also seen in some non-neoplastic conditions. To more definitively differentiate carcinomas from non-neoplastic lesions, we analyzed the stereomicroscopic structure of macroscopically granular, flat, or gastric area-like mucosal lesions with a methylene-blue contrast technique. Sixteen superficial flat early carcinomas and 65 non-neoplastic flat lesions from surgically resected gallbladders were studied by stereomicroscopy. The fine mucosal structures were classified into three patterns: grooved, pitted, or papillary, each of which was further subdivided into regular and irregular. The frequency of the grooved (52.2%) and papillary (52.2%) patterns was significantly higher in the carcinomas than in the non-neoplastic lesions (24.7% and 1.3%, respectively). The pitted pattern was present in 69.6% of the carcinomas and in 53.2% of the non-neoplastic lesions (the difference was not significant). In the grooved and pitted patterns, the irregular subtypes predominated in the carcinomas (100% and 81.3%, respectively), while the regular subtypes were more frequent in the non neoplastic lesions (84.2% and 97.6%, respectively). Stereomicroscopic examination of the fine mucosal structures of flat lesions of the gallbladder is very useful in differentiating carcinomas from non-neoplastic lesions. PMID- 9349990 TI - Gastric neuroendocrine carcinoma associated with chronic atrophic gastritis type A. AB - We report a case of gastric neuroendocrine (NE) carcinoma associated with chronic atrophic gastritis type A (CAG/A) or reversed atrophic type gastritis. A 9 x 6 cm tumor was resected from the stomach to control pain in a 55-year-old Japanese woman with peritoneal dissemination and metastatic tumors of the liver and ovary. Histologically, the tumor was NE carcinoma which showed an organoid structure, but consisted of NE cells with overt cytological atypia and frequent mitotic activity. Multiple microcarcinoids and NE cell micronests (NECMs) were also observed in the atrophic non-neoplastic mucosa of the gastric body. CEA immunoreactivity and a high Ki-67 labeling index were characteristic features of the neoplastic NE cells of the carcinoma. Although most NE tumors arising from CAG/A are typical carcinoid tumors, the present case illustrates that a high grade NE carcinoma can develop from diverse NE cell proliferation in association with CAG/A. PMID- 9349991 TI - Upper gastrointestinal bleeding arising from metastatic testicular tumor. AB - We report a young man with a testicular tumor in whom the first symptom was upper gastrointestinal bleeding. This was caused by invasion of the duodenum by a metastatic lesion in a retroperitoneal lymph node. Metastatic testicular tumor is a rare cause of upper gastrointestinal bleeding. Diagnosis was difficult because the primary lesion could not be found initially. In young men with upper gastrointestinal bleeding, the possibility of metastatic testicular tumor should be investigated. PMID- 9349992 TI - Acute lupus peritonitis successfully treated with steroid pulse therapy. AB - A 21-year-old man with systemic lupus erythematosus (SLE) who developed acute lupus peritonitis is described. Acute lupus peritonitis appeared during generalized lupus flare, with nausea, vomiting, frequent diarrhea, and abdominal tenderness with rebound and guarding. The patient was afebrile and had decreased bowel sounds. Abdominal ultrasonography and computed tomography revealed marked thickening of the gastric, duodenal, and jejunal walls, massive intraluminal fluid collection, and increasing ascites. Gastrointestinal endoscopy showed edematous mucosa with multiple erosions of the stomach and duodenum. The ascitic fluid was remarkable for low complement levels and elevated anti-DNA antibody. These manifestations of acute lupus peritonitis resolved after steroid pulse therapy with methylprednisolone. We should consider acute lupus peritonitis in a patient with SLE when abdominal symptoms are severe. Experience with our patient indicates that steroid pulse therapy is effective for this rare but severe manifestation of SLE. PMID- 9349993 TI - Primary adenocarcinoma of appendix, colonic type associated with perforating peritonitis in an elderly patient. AB - We report a case of colonic type adenocarcinoma of the appendix with perforating peritonitis in a 92-year-old man. The preoperative diagnosis was localized peritonitis due to acute appendicitis and emergency laparotomy was performed. A gray, hard tumor was palpated at the base of the appendix. Appendiceal cancer was suspected, and right hemicolectomy was performed. The histopathological diagnosis was moderately differentiated adenocarcinoma of the appendix. The tumor obstructed the orifice of the appendix, and this may have caused the perforation of the appendix. The patient had an uneventful postoperative course and there have been no signs of recurrence in the 2 years since the operation. PMID- 9349994 TI - Crohn's disease associated with renal amyloidosis successfully treated with an elemental diet. AB - We report a case of Crohn's disease associated with nephrotic syndrome due to renal amyloidosis in a 21-year-old man in whom remission of both Crohn's disease and the nephrotic syndrome has been maintained with an elemental diet. The patient developed toxic megacolon and nephrotic syndrome due to renal amyloidosis. Intensive intravenous prednisolone therapy with total parenteral nutrition was dramatically effective in treating the toxic megacolon and inducing remission in Crohn's disease and afterward, remission of the nephrotic syndrome. Remission of both conditions has been maintained for more than 2 years with the elemental diet. To our knowledge, this is the first confirmed case of Crohn's disease complicated with renal amyloidosis in which only slight proteinuria (below 0.3 g/day) was shown with an elemental diet used for a long period. PMID- 9349995 TI - Acute exacerbation of hepatitis due to reactivation of hepatitis B virus with mutations in the core region after chemotherapy for malignant lymphoma. AB - A 43-year-old Japanese man who was positive for hepatitis B surface (HBs) antigen and HB e antibody, underwent chemotherapy for non-Hodgkin's lymphoma. After the chemotherapy he suffered from acute exacerbation of hepatitis because of reactivation of HBV. Recovery was achieved with interferon-alpha, glucagon insulin therapy, and plasma exchange. Mutations were detected in codons 97, 100, 129, and 131 of the core region of HBV. The peptide encoded from the core region including such mutations possibly had greater antigenicity to induce cytotoxic T cell activity in the host. Core region mutations may be a crucial cause of the acute exacerbation of hepatitis B seen after chemotherapy. PMID- 9349996 TI - Hepatic actinomycosis: case report and review of the literature in Japan. AB - Hepatic actinomycosis is rare. We report an 86-year-old Japanese man with a 3-day history of high fever and anorexia who had an actinomycotic liver abscess complicated by disseminated intravascular coagulation (DIC). A definitive diagnosis was made when an Actinomyces species was cultured from aspirated pus. The clinical course was satisfactory. Treatment included prompt percutaneous drainage coupled with long-term intravenous administration of high-dose minocycline and piperacillin, combined with therapy for DIC. We reviewed 11 cases in Japan of Actinomyces involving the liver, including the case reported here. In most patients, there were no predisposing factors. Common symptoms and laboratory findings included fever, abdominal pain, leukocytosis, and elevated C-reactive protein. In 6 of the 11 patients a partial hepatectomy was performed because hepatic tumor was suspected. Five patients presented with a liver abscess. Hepatic actinomycosis should be considered in the differential diagnoses of pyogenic liver abscess and space-occupying lesions of the liver. PMID- 9349998 TI - Hepatic infarction with portal thrombosis. AB - A case of hepatic infarction with portal thrombosis is reported. A 63-year-old woman with liver cirrhosis and esophageal varices was admitted for treatment of the esophageal varices. Endoscopic variceal ligation (EVL) and endoscopic injection sclerotherapy (EIS) were performed. Two months later, she experienced right hypochondralgia and right flank pain. Serum transaminase levels were suddenly elevated, and computed tomography scans of the liver showed multiple small nodular lesions. Her condition worsened, and she died of hepatic failure. Autopsy revealed splenic and portal vein thrombosis, multiple hepatic infarction, and evidence of chronic pancreatitis. We believe that liver cirrhosis and chronic pancreatitis were the main risk factors for the portal thrombosis, and the treatment for esophageal varices appeared to have triggered the thrombosis. The hepatic infarction was caused by the portal thrombosis. PMID- 9349997 TI - Usefulness of novel imaging modalities in diagnosis of focal nodular hyperplasia of the liver. AB - A 17-year-old woman was admitted because of a liver tumor found incidentally by ultrasonography. Liver function was normal and there were no markers of hepatitis viruses or malignancy. Abdominal ultrasonography, computed tomography (CT), and magnetic resonance imaging revealed a mass (2 cm in diameter) in the lateral segment of the left lobe of the liver. The lesion was not detected by hepatic arteriography. However, dynamic CT with fast scanning and dynamic CO2-enhanced ultrasonography demonstrated initial central enhancement of the mass followed by centrifugal spread of enhancement to the periphery. Color Doppler flow imaging detected a central color spot, shown to be an artery by a pulsed Doppler spectrum analysis. Fine-needle biopsy confirmed a diagnosis of focal nodular hyperplasia. Dynamic CT with fast scanning, dynamic CO2-enhanced ultrasonography, and color Doppler flow imaging were useful in detecting the vascular pattern specific to focal nodular hyperplasia. Investigation of further cases with these novel imaging modalities should help to establish a comprehensive diagnostic procedure and thus avoid unnecessary surgery for focal nodular hyperplasia, which is a completely benign lesion. PMID- 9349999 TI - Exacerbated autoimmune hepatitis successfully treated with leukocytapheresis and bilirubin adsorption therapy. AB - A 58-year-old man with subacute fulminant onset of autoimmune hepatitis (AIH) was treated by leukocytapheresis (LCAP) and bilirubin adsorption therapy (BAT), rather than by administration of high-dose corticosteroids as he had mild glucose intolerance, and a definitive diagnosis of AIH was not obtained on admission; further, there was a risk of viral infection. After initiation of the therapies, serum transaminases and bilirubin, immunoglobulins, anti-nuclear antibodies, and rheumatoid factor decreased rapidly, as did the initially high levels of activated cells and several pro-inflammatory cytokines. Liver inflammation observed on liver biopsy settled during the course of the therapies, with no adverse side effects. A pause in the therapies was associated with deterioration; however, restoration of apheresis was followed by normalization. Remission was sustained throughout the period monitored, except for a recurrence 14 months after discharge, which was successfully resolved by two additional LCAP sessions. These results suggest that LCAP influences the causal mechanism(s) of exacerbation of AIH. PMID- 9350000 TI - Cowden's disease with a defined genetic alteration--chromosomal duplication at 15q11-q13. AB - Cowden's disease, multiple hamartoma syndrome, is a dominantly inherited disorder characterized by multiple hamartomas of ectodermal, endodermal, and mesodermal origin and also by a high incidence of malignant tumors. Despite many efforts to identify the genetic alterations responsible for the syndrome, the molecular mechanism remains unclear. We report a case of Cowden's disease in which karyotype analysis revealed a small duplication (about 1 Mb) at 15q11-q13. This part of the genome is a region that is deleted in the Prader-Willi/Angelman syndrome and is a "hot spot" of chromosomal duplication. PMID- 9350001 TI - Lymphoepithelial cyst of the pancreas. AB - A case of lymphoepithelial cyst (LEC) of the pancreas is presented. A 48-year-old man complaining of general fatigue was found to have a heterogeneous water-dense mass protruding from the surface of the pancreas on plain computed tomography (CT). Dynamic CT disclosed septa within the mass. Magnetic resonance imaging, (MRI) showed a hypointense mass on T1-weighted imaging, and a hyperintense mass on T2-weighted imaging. MRI with gadolinium enhancement revealed septa within the mass. Endoscopic ultrasonography showed septa and fine echogenic structures within the cystic echoic lesion. Endoscopic retrograde pancreatography showed a normal duct system. Distal pancreatectomy with splenectomy was performed, with a suspected diagnosis of cystic neoplasms of the pancreas. Histopathologic examination disclosed LEC of the pancreas. Our case suggests that LEC should be considered in the differential diagnosis of cystic neoplasms of the pancreas. PMID- 9350002 TI - Polypoid adenomyoma of the gallbladder. AB - We describe a distinctive polypoid lesion of the gallbladder (16 mm in diameter) at the fundus, associated with a granulomatous mass in the liver adjacent to the gallbladder fossa, in a 64-year-old Japanese woman. Preoperatively, the lesion was diagnosed as advanced gallbladder cancer infiltrating the liver, and hepatopancreato-duodenectomy was performed. In the resected specimen, the polyp was round and pedunculate in shape, and, microscopically, it consisted of a mixture of small glandular and surrounding muscular elements, but atypia was not noted. The constituent elements were identical with those of adenomyoma, but the polypoid appearance was unusual. The hepatic lesion proved to be a foreign body granuloma containing multiple barium fragments and giant cells. A deep gastric ulcer, which penetrated into the gallbladder fossa, was also noted, near the granuloma. The histologic features indicate that the polypoid appearance of the tumor was due to a secondary modification of pre-existing adenomyoma by hepatic granuloma. To our knowledge, this is the second reported case of an adenomyomatous lesion of the gallbladder with a polypoid appearance. PMID- 9350003 TI - Steroid hormone-dependent overexpression of cytochromes P450 2A in liver tumors of TGF alpha transgenic male mice. AB - To clarify the mechanism underlying the male preference of liver tumor in transforming growth factor (TGF) alpha transgenic mice, we analyzed the sexually dimorphic expression of two P450s, i.e., female-specific mouse 15 alpha hydroxylase P450 (2A4) and coumarin 7-hydroxylase P450 (2A5). The expression of 2A4 mRNA in the livers of both transgenic and nontransgenic males was low compared with that in females. P450 2A5 mRNA in the transgenic males was slightly elevated in the adjacent non-tumorous tissues and dramatically elevated in the tumor compared with that in nontransgenic male liver. The activity of P450 2A5 was higher in females than in males in control and transgenic mice but the difference was smaller in the transgenic mice. The activity of P450 2A5 was exceptionally high in liver tumors of transgenic males, as indicated by mRNA expression. These results suggest that female-specific P450 2A5 is induced in the livers of TGF alpha transgenic male mice, particularly in liver tumors of transgenic male mice overexpressing TGF alpha, and may be useful as a marker for mouse hepatocarcinogenesis. PMID- 9350004 TI - Myogenic cells express multiple myosin isoforms. AB - In vivo and in vitro, proliferating motile myoblasts form aligned groups of cells, with a characteristic bipolar morphology, subsequently become post mitotic, begin to express skeletal myosin and fuse. We were interested in whether members of the myosin superfamily were involved in myogenesis. We found that the myoblasts expressed multiple myosin isoforms, from at least five different classes of the myosin superfamily (classes I, II, V, VII and IX), using RT-PCR and degenerate primers to conserved regions of myosin. All of these myosin isoforms were expressed most highly in myoblasts and their expression decreased as they differentiated into mature myotubes, by RNAse protection assays, and Western analysis. However, only myosin I alpha, non-muscle myosin IIA and IIB together with actin relocalize in response to the differentiative state of the cell. In single cells, myosin I alpha was found at the leading edge, in rear microspikes and had a punctate cytoplasmic staining, and non-muscle myosin was associated with actin bundles as previously described for fibroblasts. In aligned groups of cells, all these proteins were found at the plasma membrane. Co staining for skeletal myosin II, and myosin I alpha showed that myosin I alpha also appeared to be expressed at higher levels in post-mitotic myoblasts that had begun to express skeletal myosin prior to fusion. In early myotubes, actin and non-muscle myosin IIA and IIB remained localized at the membrane. All of the other myosin isoforms we looked at, myosin V, myosin IX and a second isoform of myosin I (mouse homologue to myr2) showed a punctate cytoplasmic staining which did not change as the myoblasts differentiated. In conclusion, although we found that myoblasts express many different isoforms of the myosin superfamily, only myosin I alpha, non-muscle myosin IIA and IIB appear to play any direct role in myogenesis. PMID- 9350005 TI - The predominant defect in dilute melanocytes is in melanosome distribution and not cell shape, supporting a role for myosin V in melanosome transport. AB - Mice with mutations at the dilute locus, which encodes the heavy chain of a type V unconventional myosin, exhibit a reduction in coat colour intensity. This defect is thought to be caused by the absence in dilute melanocytes of the extensive dendritic arbor through which these cells normally deliver pigment laden melanosomes to keratinocytes. The data on which this conclusion has been based can also be explained, however, by a defect in the outward transport of melanosomes within melanocytes of normal shape. To resolve this question, we compared the shape and pigment distribution within melanocytes present in primary cultures prepared from the epidermis of C57BL/6J pups that were either wild type (D/D) at dilute or homozygous for the dilute null allele d120J. These same comparisons were also performed on melanocytes in situ, where antibodies to the membrane tyrosine kinase receptor cKIT were used to visualize melanocyte cell shape independent of pigment distribution. Wild type melanocytes were found to be dendritic and to have melanosomes distributed throughout their dendrites both in vitro and in situ. Mutant melanocytes were also found to be dendritic in both cases, but their melanosomes were highly concentrated in the cell body and largely excluded from dendrites. We conclude, therefore, that the predominant defect in dilute melanocytes is in melanosome distribution, not cell shape. These results argue that the myosin V isoform encoded by the dilute locus functions in dendritic extensions to move melanosomes from their site of formation within the cell body to their site of intercellular transfer at dendritic tips. This conclusion is consistent with our recent demonstration by immunolocalization that the dilute myosin V isoform associates with melanosomes in mouse melanocytes. PMID- 9350006 TI - Differences in myosin head arrangement on relaxed thick filaments from Lethocerus and rabbit muscles. AB - Relaxed thick filaments from insect asynchronous flight muscle appear different from those of other striated muscles, both in sections and as separated, negatively-stained structures. Unlike relaxed filaments of scallops, chelicerate arthropods, or vertebrate striated muscle, all of which display a predominantly helical arrangement of surface myosin heads, insect asynchronous flight muscle filaments appear striped, with cross-striations or shelves at spacings of 14.5 nm. Using a bifunctional agent to cross-link the active sites of nearest neighbour myosin heads we previously demonstrated that the helical arrays on the surfaces of scallop, arthropod, fish and frog filaments are produced by the association of two oppositely-oriented myosin heads, each of which originates from an axially sequential molecule within the same helical strand. The effect of similarly cross-linking nearest-neighbour heads with the bifunctional agent 3,3' dithiobis[3'(2')-O-(6-propionylamino)hexanoyl]adenosine 5'-triphosphate in the presence of vanadate on the solubility of thick filaments separated from Lethocerus indirect flight muscle (an insect asynchronous flight muscle) and rabbit psoas muscle was examined. After incubation on high salt, treated rabbit filaments retained their length and surface myosin, while untreated filaments and those with severed cross-links dissolved, indicating that the myosin head arrangement on rabbit filaments is similar to those previously studied. Treated indirect flight muscles filaments, however, separated into distinct segments of variable lengths, usually multiples of 150 nm, while untreated filaments and those with severed cross-links dissolved completely. This implies that intermolecular associations on indirect flight muscles filaments most likely occur between circumferentially-adjacent heads within each crown, but originating from different helical strands. We interpret this difference in the relaxed orientations of splayed myosin heads on the two types of filament as reflecting a difference in functional requirements at the onset of, or during, contractile activity. PMID- 9350007 TI - Coordinated fast-to-slow transitions of myosin and SERCA isoforms in chronically stimulated muscles of euthyroid and hyperthyroid rabbits. AB - Changes in the patterns of myosin heavy chain (MHC) isoforms, isomyosins, and Ca(2+)-ATPase (SERCA) isoforms were studied in long-term (72 d) stimulated fast twitch extensor digitorum longus (EDL) and tibialis anterior (TA) muscles of euthyroid and hyperthyroid rabbits. The chronic low-frequency stimulation-induced fast-to-slow transitions in MHC isoforms, isomyosins and SERCA isoforms were pronounced in muscles from euthyroid rabbits, but less pronounced in muscles from hyperthyroid rabbits. Thus, hyperthyroidism counteracted to same extent the stimulation-induced fast-to-slow transition. Analyses of all parameters were performed on the same individual muscles, providing information on the co ordinated expression of SERCA and myosin isoforms. A high correlation (r = 0.97) was detected between relative concentrations of slow SERCA2a and slow MHCI isoforms. This correlation persisted under all experimental conditions, suggesting a co-ordinated expression of slow myosin and Ca(2+)-ATPase isoforms. Conversely, fast SERCA1a was correlated to fast myosin isoforms as a whole. PMID- 9350008 TI - Stretch activation and isoforms of myosin heavy chain and troponin-T of rat skeletal muscle fibres. AB - Recent studies on single mammalian skeletal muscle fibres revealed a correlation between the kinetics of stretch-induced delayed force increase (stretch activation) and the isoforms of the myosin heavy chain. This observation suggests a causal relation between stretch activation and myosin heavy chain. However, the assumption is weakened by the fact that isoforms of other myofibrillar proteins tend to be coexpressed with myosin heavy chain isoforms. The relation between the isoforms of the tropomyosin-binding troponin subunit and myosin heavy chain is unknown. For a variety of reasons, tropomyosin-binding troponin subunit is a possible candidate for being involved in stretch activation. Therefore, we measured stretch activation of single, maximally Ca(2+)-activated skinned rat skeletal muscle fibres and characterized them by their myosin heavy chain composition, as well as by the isoform species of tropomyosin-binding troponin subunit. Four myosin heavy chain isoforms (I, IIa, IId or IIx and IIb) and six tropomyosin-binding troponin subunit isoforms (TnT1s, TnT2s, TnT1f, TnT2f, TnT3f, TnT4f) were distinguished. The following preferential coexpression patterns of the myosin heavy chain and tropomyosin-binding troponin subunit isoforms were observed: MHCI-TnT1s, MHCIIa-TnT3f, MHCIId-TnT1f, and MHCIIb-TnT4f. Stretch activation kinetics was found to be correlated with the myosin heavy chain isoform complement also in fibres not displaying one of the preferential MHC-TnTf isoform coexpression patterns. This corroborates the assumption of a causal relation between myosin heavy chain and stretch activation. PMID- 9350009 TI - Nucleotide and actin binding properties of the isolated motor domain from Dictyostelium discoideum myosin. AB - Nucleotide and actin binding properties of the truncated myosin head (S1dC) from Dictyostelium myosin II were studied in solution using rabbit skeletal myosin subfragment 1 as a reference material. S1dC and subfragment 1 had similar affinities for ADP analogues, epsilon ADP and TNP-ADP. The complexes of epsilon ADP and BeFx or AIF4- were less stable with S1dC than with subfragment 1. Stern Volmer constants for acrylamide quenching of S1dC complexes with epsilon ADP, epsilon ADP.AIF4- and epsilon ADP.BeFx were 2.6, 2.9 and 2.2 M-1, respectively. The corresponding values for subfragment 1 were 2.6, 1.5 and 1.1 M-1. The environment of the nucleotide binding site was probed by using a hydrophobic fluorescent probe, PPBA. PPBA was a competitive inhibitor of S1dC Ca(2+)-ATPase (Ki = 1.6 microM). The binding of nucleotides to subfragment 1 enhanced PPBA fluorescence and caused blue shifts in the wavelength of its maximum emission in the order: ATP approximately ADP.AIF4- approximately ADP.BeFx > ATP gamma S > ADP > PPi. In the case of S1dC, the effects of different nucleotides were smaller and indistinguishable from each other. S1dC bound actin tighter than S1 (Kd = 7 nM and 60 nM, respectively). The actin activated MgATPase activity of S1dC varied between preparations, and the Vmax and K(m) values ranged between 3 and 7 s-1 and 60 and 190 microM, respectively. S1dC showed lower structural stability than S1 as revealed by their thermal inactivations at 35 degrees C. These results show that the nucleotide and actin binding of S1dC and subfragment 1 are similar but there are some differences in nucleotide and phosphate analogue-induced changes and the communication between the nucleotide and actin binding sites in these proteins. PMID- 9350011 TI - Expression of ion channels during differentiation of a human skeletal muscle cell line. AB - An immortal, cloned cell line (RCMH), obtained from human skeletal muscle was established in our laboratory and shown to express muscle specific proteins. We measured ligand binding to ion channels, ion currents using whole cell patch clamp and intracellular calcium both in cells grown in complete media and in cells grown for 4-40 days in media supplemented with hormones and nutrients (differentiating media). Markers for differentiated muscle, such as the muscle isoform of creatine kinase and the cytoskeletal proteins alpha-actinin, alpha sarcomeric actin, myosin and titin were present in early stages. Receptors for gamma toxin from Tityus serrulatus scorpion venom, a specific modulator for voltage dependent sodium channels, were present (0.9-1.0 pmol mg-1 protein) during stage 1 (0-6 days in culture with differentiating media) and increased by 50% in stage 3 (more than 10 days in differentiating media). High and low affinity dihydropyridine receptors present in stage 1 convert into a single type of high affinity receptors in stage 3. Both intracellular calcium release and InsP3 receptors were evident in stage 1 but ryanodine receptors were expressed only in stage 3. RCMH cells showed no voltage sensitive currents in stage 1. Between 7 and 10 days in differentiating media (stage 2), an outward potassium current was observed. Small inward currents appeared only in stage 3; we identified both tetrodotoxin sensitive and tetrodotoxin resistant sodium currents as well as calcium currents. This pattern is consistent with the expression of voltage dependent calcium release before appearance of both the action potential and ryanodine receptors. PMID- 9350010 TI - Effects of long-term phasic electrical stimulation on denervated soleus muscle: guinea-pig contrasted with rat. AB - Guinea-pig soleus muscles were denervated and electrically stimulated for periods of 43 to 66 days. Stimuli were in 1 s bursts of 40 Hz pulses, repeated every 5 min. Other guinea-pigs were denervated for 82 days without stimulation and, in a third group, the soleus muscle was necrotized and allowed to regenerate without reinnervation for 13-15 days. Isometric and isotonic recordings were made in vivo. Denervated guinea-pig muscles were embedded in epoxy resin for light and electron microscopy. Chronic stimulation of denervated guinea-pig soleus had no effects on the prolonged twitch or on reduced maximal shortening velocity, maximal rate of rise of tension and tetanic force. This contrasts with the slow to-fast conversion produced by denervation and denervation-stimulation of rat soleus. Loss of force was much greater in rat than guinea-pig after denervation, and chronic stimulation increased force in rat to the same level as in guinea-pig after denervation (with or without stimulation). Eighty-day denervated guinea-pig soleus did not reveal those morphological signs of fibre breakdown and regeneration which are prominent in denervated rat soleus muscles. Those changes in rat resembled aneurally regenerated muscles in several aspects, especially the increased incidence of fibres with internal myo-nuclei which did not appear in guinea-pig soleus after denervation. Aneurally regenerated guinea-pig soleus became fast like aneurally regenerated rat muscle. Our data are compatible with the hypothesis that slow-to-fast transformation of denervated rat soleus is not directly brought about by chronic stimulation but by de-novo formation of fast contracting regenerated fibres. The persistence of fibrillation in guinea-pig but not rat after denervation may account for the species difference. PMID- 9350013 TI - Endosalpingiosis and chronic pelvic pain. AB - OBJECTIVE: To show that endosalpingiosis (ES) is a frequently underdiagnosed or misdiagnosed entity in women thought to have endometriosis (EM) on initial pathologic review and that ES alone may be an independent cause of pelvic pain. STUDY DESIGN: A retro-spective review of pathology records from gynecologic cases from June 1992 to November 1994 revealed 37 cases of EM and 5 cases of ES. These cases were reviewed by a single pathologist (J.M.), who assigned a final diagnosis. Preoperative diagnosis or symptoms that led to surgery were compared to the initial and final pathologic diagnoses. RESULTS: Of the 37 cases with the initial diagnosis of EM, 64.9% had EM only (group I), 18.9% had both EM and ES (group II), and 16.2% had ES alone (group III). Of the five patients with an initial diagnosis of ES, 80% had ES (group IV), while 20% had both ES and EM (group V). In patients who had ES as their final diagnosis (group III and IV), 70% had chronic pelvic pain or presumed endometriosis as a preoperative diagnosis. This was similar to the 53.1% of patients with a final diagnosis of EM who had the same preoperative diagnosis. CONCLUSION: We conclude that ES is more common than once thought. Furthermore, when an initial diagnosis of EM is made, further review may reveal that many of these cases are a combination of both EM and ES, while still others may be ES alone. ES, which has been long been thought to be clinically insignificant, could be a source of pelvic pain. PMID- 9350012 TI - The effect of pH on the Ca2+ affinity of the Ca2+ regulatory sites of skeletal and cardiac troponin C in skinned muscle fibres. AB - It is known that intracellular pH drops rapidly after the onset of ischemia in cardiac muscle and may play some role in the rapid drop in force that ensues. It is also known that alpha 1-adrenoceptor agonists alkalinize intracellular pH by stimulating Na+/H+ exchange and may represent a mechanism which facilitates recovery of intracellular pH from acidosis. Lowering or raising pH shifts the Ca2+ dependence of force development in muscle fibres to higher or lower free Ca2+ concentrations, respectively, yet the precise mechanism is unknown. To investigate this phenomenon we have used skinned skeletal or cardiac muscle fibres whose endogenous troponin C (TnC) has been replaced with chicken skeletal TnC labelled with DANZ (STnCDANZ) or recombinant cardiac TnC labelled with IAANS (CTnC3(C84)[AANS), respectively. The fluorescence of the STnCDANZ or CTnC3(C84)IAANS was enhanced by Ca2+ binding to the Ca(2+)-specific (regulatory) site(s) of STnC or CTnC when incorporated into skinned fibres, and was measured simultaneously with force. When the pH was changed from 7.0 to 6.5 or 7.5 the shift in the Ca2+ dependence of force paralleled the shift in fluorescence. Since the force and fluorescence shift in parallel as the pH is lowered or raised, it can be concluded that these changes in Ca2+ sensitivity are caused by a decrease or increase, respectively, in the Ca2+ affinity of the Ca(2+)-specific site(s) of TnC. Since lowering or raising the pH also resulted in lower or higher, respectively, maximal Ca2+ activated force while maximal fluorescence remained unchanged, it is possible that H+ may act indirectly, as well, by reducing or increasing, respectively, the number or type of crossbridges attached to actin and thereby alter the crossbridge induced depression or elevation, respectively of the observed TnC Ca2+ affinity. Experiments with 2,3-butanedione monoxime, however, where force-generating crossbridges were greatly reduced, indicated that the pH effect may be primarily related to a direct change in the Ca2+ affinity to the regulatory sites of TnC. PMID- 9350014 TI - Serum CA-125 measurements > 65 U/mL. Clinical value. AB - OBJECTIVE: To review the prevalence of various conditions associated with serum CA-125 values > 65 U/mL, to calculate the odds ratios of different ranges of high CA-125 in predicting cancer and to study the effect of menopause and the presence of a mass on the predictive value of high serum CA-125. STUDY DESIGN: A retrospective review of the diagnoses in 313 consecutive women seen at the Cleveland Clinic Foundation whose serum CA-125 was > 65 U/mL was performed. Statistical analysis was performed using crosstabulation, chi 2, Fisher's exact test and the odds ratio. RESULTS: In patients with serum CA-125 > 65 U/mL, gynecologic cancers, nongynecologic cancers and non-malignant conditions constituted 74.3%, 10.2% and 13.1% of diagnoses, respectively. In patients with serum CA-125 > or = 1,000 U/mL, the same conditions were responsible for 89%, 7% and 3% of diagnoses, respectively. Endometriosis and metastatic breast cancer were the most common benign condition and nongynecologic cancer associated with serum CA-125 > 65 U/mL. The presence of an abdominopelvic mass significantly increased the risk of malignancy (P < .00005). Approximately 90% of patients with CA-125 > 65 U/mL and no mass had nonmalignant disease. The diagnoses of serum CA 125 values > 65 U/mL varied significantly in premenopausal versus postmenopausal patients. Postmenopausal patients had a higher incidence of gynecologic (P = .002) and nongynecologic (P = .0008) cancers and lower incidence of benign conditions (P < .0005). The odds ratio that CA-125 levels were associated with cancer increased as the level of CA-125 increased. The odds ratio of malignant versus benign disease was significantly higher in post-menopausal patients for all intervals of CA-125 levels until the level of > or = 1,000 U/mL was reached. CONCLUSION: In patients seen at a tertiary center, serum CA-125 measurements > 65 U/mL were associated with nonmalignant conditions in 13% of patients. Although higher serum CA-125 levels were more associated with gynecologic malignancies, no level of CA-125 occurred exclusively with gynecologic cancers. In postmenopausal patients with serum CA-125 values > 65 U/mL and in patients with serum CA-125 values > 65 U/mL and an abdominopelvic mass, subspecialty consultation should be considered before proceeding to surgery. PMID- 9350015 TI - Laparotomy sponge adhesions over the uterus. PMID- 9350016 TI - High-risk obstetrics. PMID- 9350018 TI - Patient anxiety during gynecologic examinations. Behavioral indicators. AB - OBJECTIVE: To identify behaviors that indicate anxiety during a gynecologic examination. STUDY DESIGN: Five hundred twenty-two women visiting a private obstetrician/gynecologist's office completed the A-State scale of the State-Trait Anxiety Inventory and specific questions about their first pelvic examination and experiences with health practitioners performing subsequent gynecologic examinations. In addition, the hand placement a woman exhibited as the speculum was inserted was recorded, as were the reasons for her visit, reports of any symptoms, performance of any special procedures (e.g., colposcopy) and whether the pelvic examination was her first. RESULTS: Five behaviors observed during speculum insertion--holding hands/eyes covered or shut, hands on shoulders, hands covering pelvis, hands on legs, hands holding table--indicated increased anxiety. Together these behaviors were exhibited by one of every four patients and were found to be associated with high levels of anxiety. Greater anxiety was related to colposcopy, a less positive first pelvic examination experience, overall less positive experiences with examiners and performance of the first gynecologic examination at the present visit. CONCLUSION: Easily recognizable behaviors reflecting high anxiety in gynecologic patients were identified. Upon recognizing these behaviors, examiners can take necessary measures to reduce patient anxiety and prevent delays in and avoidance of gynecologic examinations. PMID- 9350017 TI - Closure of Pfannenstiel skin incisions. Staples vs. subcuticular suture. AB - OBJECTIVE: To compare skin closure with staples and subcuticular suture. STUDY DESIGN: Obstetric patients undergoing cesarean section with a Pfannenstiel incision were prospectively randomized to skin closure with staples or subcuticular suture. Pain and cosmesis were assessed postoperatively. RESULTS: Patients reported significantly less pain following subcuticular closure at both the time of discharge (P < or = .01) and the postoperative visit (P < or = .002). Incisions closed with subcuticular suture were found to be more cosmetically attractive by both patients (P = .04) and physicians (P = .01) at the postoperative visit. CONCLUSION: Pfannenstiel skin incisions closed with subcuticular closure following cesarean section result in less postoperative discomfort and are more cosmetically appealing at the six-week postoperative visit as compared to incisions closed with staples. PMID- 9350019 TI - Premenstrual symptoms in general practice patients. Prevalence and treatment. AB - OBJECTIVE: To examine the rates of premenstrual symptoms in Australian patients, the treatments they had tried for such symptoms, the perceived effectiveness of these treatments, the proportion of women who reported that they had sought help for premenstrual symptoms and whether women perceived the need for additional help in dealing with premenstrual symptoms. Characteristics associated with higher symptom levels and desire for help were examined. STUDY DESIGN: A cross sectional survey of 310 general practices patients aged 18-45 years and who had reported having had a menstrual period in the previous three months. RESULTS: Between 11% and 32% of women reported severe or extreme changes during the premenstrual phase on each of the 10 symptoms in the short Premenstrual Assessment Form, with the highest rates for affective symptoms. Eighty-five percent of women reported that they had tried treatments for premenstrual symptoms, and many reported having tried multiple treatments. The most commonly tried treatments included pain killers rest, drinking more fluid and exercise, which had been tried by at least one-third of women. When women were asked to nominate up to three treatments they had tried and found most effective, the most commonly mentioned were dietary changes, evening primrose oil, vitamins (including B6) and exercise. Approximately 50% of the women had sought help, most commonly from a general practitioner and 45% reported that they would like more help dealing with premenstrual symptoms. Higher overall symptom scores were associated with a history of endometriosis, a lower education level, not taking oral contraceptives, taking evening primrose oil and taking vitamin B6. CONCLUSION: There is a need to further refine, through evaluation of different approaches, programs and resources, ways to effectively help women who report premenstrual symptoms and would like help to deal with them. PMID- 9350020 TI - Interpreting fetal heart rate tracings. Is there a difference between labor and delivery nurses and obstetricians? AB - OBJECTIVE: To analyze the influence of increasing education and clinical experience on fetal heart rate interpretation by health care providers in labor and delivery areas. STUDY DESIGN: Eleven tracings representing a variety of fetal heart rate patterns and neonatal outcomes were selected. Respondents were asked to interpret the tracing strips on a 1-5 scale and predict Apgar scores and cord blood gases. RESULTS: Seventy nurses and physicians participated. Experience with labor and delivery and provider classification correlated significantly with correct interpretation. Certified nurse midwives differed significantly in number of correct interpretations from both first-year residents and low-risk nurses. Providers did not differ on ability to predict Apgar scores or cord gases. CONCLUSION: Length of clinical experience correlated positively with the interpretation of fetal heart rate tracings but not with the prediction of Apgar scores or cord blood gas measurements. These data will be useful in designing educational and orientation programs for physicians and nurses in the labor and delivery setting. PMID- 9350022 TI - Logbook data as a source of birth information. Are they valid? AB - OBJECTIVE: To determine if the data recorded in the labor and delivery logbook in two hospitals are complete and consistent with the information in the patient's medical record. STUDY DESIGN: We performed a retrospective comparison of the information in the labor and delivery logbook to the content of 110 patients' hospital charts in each of two hospitals. RESULTS: The logbooks of both hospitals had erroneous entries and missing data as compared to the antepartum complications, intrapartum events and immediate newborn care recorded in the patients' medical records. The range of error in these three categories was 61%, 31% and 60% in one hospital and 37%, 29% and 30% in the other hospital. CONCLUSION: The results of this study do not support the use of logbooks as a source of data for individual or collated departmental, hospital or agency reports containing antepartum, intrapartum and newborn information. PMID- 9350021 TI - Sexual behavior and contraceptive use. Changes from 1975 to 1995 in college women. AB - OBJECTIVE: The objective of this study was to compare the sexual practices and contraceptive use in a sample of college women in 1995 with women surveyed in 1975, 1986 and 1989. STUDY DESIGN: We surveyed 336 college women seen at a university student health service or on campus and compared their responses to those of women surveyed at the university in 1975, 1986 and 1989. RESULTS: The proportions of women who were sexually experienced, number of life-time male sexual partners, number of male sexual partners in the past year and frequencies of specific sexual practices were similar over the four survey times. Condom use was reported as the usual method of contraception in 7% of sexually experienced women in 1975, 14% in 1986, 25% in 1989 and 46% in 1995 (P < .00001, linear trend). CONCLUSION: We found little change in sexual practices in this college population over the four survey years, with the exception of an increase in the self-reported use of condoms. Increased educational efforts should emphasize safe sexual practices (barrier methods) to prevent sexually transmitted diseases and highly efficacious methods of contraception (hormonal contraception) to avoid unintended pregnancy. PMID- 9350023 TI - Long GnRH-agonist protocol in an IVF program. Is it appropriate for women with normal FSH levels and high FSH/LH ratios? AB - OBJECTIVE: To determine whether subjects with an elevated ratio of follicle stimulating hormone (FSH) to luteinizing hormone (LH) but normal basal FSH levels should be regarded as poor responders to controlled ovarian hyperstimulation. STUDY DESIGN: One hundred twenty-five women undergoing in vitro fertilization (IVF) for the first time were recruited in this retrospective cohort study. Women over 40 years old or having serum basal FSH > 10 mIU/mL were excluded. RESULTS: Various cutoff values for the FSH/LH ratio were chosen, and the ratio demonstrated that pregnancy rates were apparently higher in patients with the long protocol than with the short one if they had an FSH/LH ratio < 3.0 (48.5% vs. 25.8%, P = .034), < 2.5 (53.3% vs. 28.6%, P = .030) or < 2.0 (57.8% vs. 21.7%, P = .005). Pregnancy rates were similar with the long and short protocols in patients with FSH/LH > or = 3.0 (57.1% vs. 70%, P = .521), FSH/LH > or = 2.5 (40% vs. 53.8%, P = .435) or FSH/LH > or = 2.0 (40% vs. 55.6%, P = .281). CONCLUSION: This study failed to demonstrate that FSH/LH was a useful parameter for predicting reproductive outcome in IVF programs and for patient selection for the long or short gonadotropin-releasing hormone agonist protocol. PMID- 9350024 TI - Ovarian abscess 15 months after vaginal hysterectomy. A case report. AB - BACKGROUND: Ovarian abscess is a primary infection of ovarian parenchyma. Since 1869, only 44 cases after vaginal hysterectomy have been reported in the medical literature. The pathophysiology of bacterial infection in these cases is different from the traditional ascending mechanism. CASE: A 28-year-old woman presented with complaints of lower abdominal pain and fever 15 months after transvaginal hysterectomy. Her white blood cell count was 22,700/mm3, with 90% neutrophils. Bimanual examination revealed a tender mass in the cul-de-sac, and computed tomography showed a large, multiloculated pelvic mass. Laparotomy, pathologic examination and microbiologic study confirmed the diagnosis of ovarian abscess. CONCLUSION: Our case represents another rare posthysterectomy ovarian abscess. Most of these cases were managed by surgery and antibiotic treatment. PMID- 9350025 TI - Aplastic anemia. Report of a case with recurrent episodes in consecutive pregnancies. AB - BACKGROUND: Aplastic anemia complicating normal pregnancy is a rare event, associated with increased risks for both mother and fetus. CASE: A 30-year-old woman in her fifth pregnancy experienced the complications of aplastic anemia and preeclampsia. The patient had a history of four unsuccessful previous pregnancies also complicated by aplastic anemia, but she was free of the disease in the intervals between the pregnancies. During this last pregnancy she presented at 31 weeks' gestation with symptoms and signs of severe aplastic anemia and preeclampsia. A cesarean section was performed, and a normal infant was born. Supportive therapy, growth factors, prednisolone and intravenous immunoglobulin were administered, and the patient's general condition improved two weeks after delivery. CONCLUSION: Aplastic anemia may occur during consecutive pregnancies. The gestational age at the onset of the disease and the severity of the symptoms determine the outcome of the pregnancy. PMID- 9350026 TI - Combined tubal and cornual pregnancy in a patient without risk factors. A case report. AB - BACKGROUND: Ectopic pregnancy is the leading cause of pregnancy-related death during the first trimester. Bilateral ectopic pregnancy is a rare phenomenon, varying in frequency between 1 per 725 and 1 per 1,580 ectopic pregnancies. We report the case of a bilateral ectopic pregnancy (ruptured right cornual and intact left ampullary) in a patient with no known risk factors for extrauterine gestation. CASE: A 33-year-old, black woman, gravida 2, para 1001, presented at approximately 7 weeks' gestation with the acute onset of abdominal pain. She had a rigid surgical abdomen but was hemodynamically stable. Her beta-human chorionic gonadotropin level was 6,398 mIU/mL, and transvaginal ultrasound failed to reveal an intrauterine gestation, adnexal mass or cul-de-sac fluid. Findings at laparotomy included a 500-mL hemoperitoneum and a ruptured right cornual and intact left ampullary pregnancy. Pathology of both specimens confirmed the presence of chorionic villi. CONCLUSION: Although rare, heterotopic pregnancies can occur even in patients without risk factors. PMID- 9350027 TI - CETP and exchangeable apoproteins: common features in lipid binding activity. AB - In order to define the active domain for lipid binding in CETP (cholesteryl ester transfer protein), our study discusses some fundamental physicochemical properties of this molecule such as hydrophobic moment, protein active surface and helix amphipathicity, in comparison to the properties reported for a series of apoproteins including apoAI, apoAII, apoCI, CII, CIII and apoE. Our study suggests that CETP corresponds to a protein with an active surface slightly lower than the one calculated for the exchangeable apoproteins AI, AII, CI, CII, CIII and E. Arrays type (i, i + 3) and (i, i + 4) were found in the region associated to lipid binding in these apoproteins. Seven such arrays located in the amphipathic alpha-helices of CETP are also suggested to contribute to the overall lipid binding activity as a consequence of alpha-helix stability. It is proposed that for lipid binding to occur in both types of molecules, the possibility of a conformational specificity given by a redundant stereochemical code can be actively operating. PMID- 9350028 TI - Cloning and interspecies comparisons of three newt (Notophthalmus viridescens) fibroblast growth factor receptor sequences. AB - We report the nucleotide sequences of two fibroblast growth factor receptor (FGFR) cDNAs, FGFR1 and FGFR3, from the newt species Notophthalmus viridescens. These two cDNA sequences and a previously published newt FGFR cDNA, FGFR2, were used to derive the amino acid sequences which were then compared with their homologues from other species. This comparison shows that the intracellular tyrosine kinase domain is highly conserved across the species examined with the second half of the domain slightly more conserved than the first half. The 3' portion of the carboxyl terminal tail is not very highly conserved. The comparison of the extracellular portion of FGFR2 shows a high degree of conservation among the Ig-like domains and a low degree of conservation in the region that links the third Ig-like domain with the transmembrane domain. PMID- 9350029 TI - Ribonuclease activity dependent cytotoxicity of Asp fl, a major allergen of A. fumigatus. AB - A major allergen/antigen, Asp fl, secreted by Aspergillus fumigatus exhibits cytotoxicity towards eukaryotic cell lines. Asp fl inhibited protein synthesis in RAW cells with an IC50 of 4.5 nM and also degraded ribosomal RNA of RAW cells at a similar concentration. Ribosomal inactivation by Asp fl may be the probable mechanism for protein synthesis inhibition. Specific ribonuclease activity of Asp fl was observed to be 100,000 U/mg. Presence of strong RNase activity in Asp fl was further confirmed by agar gels containing yeast RNA. Electrophoretic run on agarose gels showed that Asp fl degrades all species of naked RNA. Modification of histidine residues of Asp fl with diethyl pyrocarbonate and alkylation of cysteines with iodoacetamide resulted in loss of ribonuclease activity and cytotoxicity of Asp fl. The current study establishes the ribonuclease activity of a purified major allergen of A. fumigatus that inhibits protein synthesis and kills the eukaryotic cells. PMID- 9350031 TI - Mitochondrial, but not peroxisomal, beta-oxidation of fatty acids is conserved in coenzyme A-deficient rat liver. AB - Hepatic coenzyme A (CoA) plays an important role in cellular lipid metabolism. Because mitochondria and peroxisomes represent the two major subcellular sites of lipid metabolism, the present study was designed to investigate the specific impact of hepatic CoA deficiency on peroxisomal as well as mitochondrial beta oxidation of fatty acids. CoA deficiency (47% decrease in free CoA and 23% decrease in total CoA) was produced by maintaining weanling male Sprague-Dawley rats on a semipurified diet deficient in pantothenic acid (the precursor of CoA) for 5 weeks. Hepatic mitochondrial fatty acid oxidation of short-chain and long chain fatty acids were not significantly different between control and CoA deficient rats. Conversely, peroxisomal beta-oxidation was significantly diminished (38% inhibition) in livers of CoA-deficient rats compared to control animals. Peroxisomal beta-oxidation was restored to normal levels when hepatic CoA was replenished. It is postulated that since the role of hepatic mitochondrial beta-oxidation is energy production while peroxisomal beta oxidation acts mainly as a detoxification system, the mitochondrial pathway of beta-oxidation is spared at the expense of the peroxisomal pathway when liver CoA plummets. The present study may offer an animal model to investigate mechanisms involved in peroxisomal diseases. PMID- 9350030 TI - Absence of endothelin receptors and receptor mRNA in mammalian fibroblasts transformed with SV40 or ras oncogene. AB - Endothelin-1 (ET-1), a peptide isolated from the culture medium of endothelial cells, mediates a variety of physiological and pathological responses including mitogenesis. We have compared the expression of ET receptors in untransformed versus ras-transformed NIH-3T3 murine fibroblasts and in untransformed versus SV40-transformed W138 (VA13) human fibroblasts by ligand binding and Northern analysis. NIH-3T3 and W138 cells displayed high affinity (200 and 220 pM) and high density (23,000 sites/cell and 14,000 sites/cell for NIH-3T3 and W138 cells, respectively) ET receptors. Competition binding experiments using subtype selective ligands identified these receptors as the ETA subtype. Addition of ET-1 to the cells produced a concentration-dependent increase in intracellular calcium release. Both ras-transformed NIH-3T3 cells and SV40-transformed W138 cells (VA13) completely lacked [125I]ET-1 binding and failed to release calcium when exposed to ET-1. Northern analysis of the polyadenylated RNA (polyA RNA) isolated from untransformed and transformed cells revealed that the steady-state level of ETA receptor RNA was 90-95% less in transformed cells compared to untransformed cells. Thus, the loss of ET receptors as well as the receptor-mediated responses in transformed cells can be explained by down-regulation of ET receptor mRNA. PMID- 9350032 TI - Influence of retinol deficiency and curcumin/turmeric feeding on tissue microsomal membrane lipid peroxidation and fatty acids in rats. AB - The effect of retinol deficiency and curcumin/turmeric on lipid peroxidation and fatty acid profile was studied in liver, kidney, spleen and brain microsomes of rats. Results revealed an increase in lipid peroxidation in retinol deficient liver by 32%, kidney 30%, spleen 24% and brain 43% compared to the controls. Feeding 0.1% curcumin or turmeric for three weeks in diet to retinol deficient rats reduced the lipid peroxidation respectively to 12.5 or 22.6%, in liver, 23.7 or 24.1% in kidney, 14.4 or 18.0% in spleen and 16.0 or 31.4% in brain. Retinol deficiency lead to a reduction in the essential fatty acids. In liver C18:1 showed a reduction by 45.6%, C18:2 by 31.6% and C20:4 by 22.8%. In kidney C18:1 was reduced by 33.6%, 18:2 by 24.6% and 20:4 by 13.7%. In spleen and brain C18:1 showed a reduction by 10.2% and 33.9%, C18:2 by 37.9% and 12.1% and C20:4 by 15.7% and 35.3% respectively. Curcumin and turmeric fed group showed a significant increase in the abnormally reduced fatty acid levels. PMID- 9350033 TI - Influence of dietary capsaicin and onion on the metabolic abnormalities associated with streptozotocin induced diabetes mellitus. AB - Effect of feeding 15 mg% capsaicin diet or 3% freeze dried onion powder containing diet were examined in albino rats rendered diabetic with streptozotocin injection. Diabetic rats maintained on onion diet for 8 weeks excreted comparatively less amounts of albumin, urea, creatinine and inorganic phosphorus. Dietary onion also partially reversed the abnormalities in plasma albumin, urea, creatinine and inorganic phosphorus in diabetic animals. Onion also produced a significant reduction in hyperglycemic status of diabetic animals. Diabetic rats maintained on onion diet had a lowered relative liver weight at the end of the study compared to diabetic control group. Diabetic rats fed onion diet also exhibited lowered lipid peroxides in circulation and in urine when compared to diabetic control group. Blood cholesterol was lowered significantly by dietary onion in diabetic animals. Cholesterol decrease was exclusively from LDL-VLDL fraction. Significant decrease in blood phospholipids and triglycerides also brought about by dietary onion. Hepatic cholesterol, triglycerides, phospholipids which were elevated under diabetic condition were countered significantly by dietary onion. Dietary capsaicin did not have any significant influence on any of the parameters tested in diabetic rats. Thus, the study reveals that onion feeding improves the metabolic status in diabetic condition, probably because of its hypoglycemic as well as hypocholesterolemic effect. PMID- 9350034 TI - 27Al-NMR studies of aluminum transport across yeast cell membranes. AB - Aluminum (Al) transport across yeast cells was studied using Dy(NO3)3 as a shift reagent by 27Al-NMR spectroscopy. The results showed that (a) Al enters the yeast cells at 15 min and over a period of time, within 4 h, an equilibrium sets in between outside and inside Al; (b) citrate does not favor Al going into the yeast cells at pH 5.0; and (c) EDTA brings out all the Al that has entered the yeast cells. PMID- 9350035 TI - Salubrious effect of Semecarpus anacardium against lipid peroxidative changes in adjuvant arthritis studied in rats. AB - Oxygen derived free radicals are known to play an important role in the etiology of tissue injury in rheumatoid arthritis. The effect of milk extract of Semecarpus anacardium nuts at the dose level of 150 mg/kg body weight for 14 days on adjuvant arthritis was studied for gaining insight into the intrigue disease in relation to the lipid peroxidation and antioxidant defence system. Increased lipid peroxides' levels in both plasma and tissues (liver, kidney and heart) of adjuvant arthritis was significantly decreased by the administration of the drug. The antioxidant defence system studied in tissues of arthritic animals were altered significantly as evidenced by the decreased level of non-enzymatic antioxidants (GSH, vitamin E, vitamin C, NPSH and TSH) and enzymatic antioxidants (catalase and GPx except SOD). Administration of Semecarpus anacardium nut extract brings back the altered antioxidant defence components to near normal levels. These observations suggest that the diseased state of adjuvant arthritis may be associated with augmented lipid peroxidation and the administration of the drug may exert its antiarthritic effect by retarding lipid peroxidation and causing a modulation in cellular antioxidant defence system. PMID- 9350036 TI - Phosphatidylserine translocation into brain mitochondria: involvement of a fusogenic protein associated with mitochondrial membranes. AB - Data reported in the literature indicate that lipid movement between intracellular organelles can occur through contacts and close physical association of membranes (Vance, J.E. 1990. J Biol Chem 265: 7248-7256). The advantage of this mechanism is that the direct interaction of membranes provides the translocation event without the involvement of lipid-transport systems. However, pre-requisite for the functioning of this machinery is the presence of protein factors controlling membrane association and fusion. In the present work we have found that liposomes fuse to mitochondria at acidic pH and that the pre treatment of mitochondria with pronase inhibits the fusogenic activity. Mixing of 14C-phosphatidylserine (PS) labeled liposomes with mitochondria at pH 6.0 results in the translocation of 14C-PS into mitochondria and in its decarboxylation to 14C-phosphatidylethanolamine through the PS decarboxylase activity localized on the outer surface of the inner mitochondrial membrane. Incorporation of 14C-PS is inhibited by the pre-treatment of mitochondria with pronase or with EEDQ, a reagent for the derivatization of the protonated form of carboxylic groups. These results indicate the presence of a protein associated with mitochondria which is able to trigger the fusion of liposomes to the mitochondrial membrane. A partial purification of a mitochondrial fusogenic glycoprotein is described in this work. The activity of the fusogenic protein appears to be dependent on the extent of protonation of the residual carboxylic groups and is influenced by the glucidic moiety, as demonstrated by its interaction with Concanavalin A. The purified protein is able to promote the recover of the 14C-PS import from liposomes to pronase-treated mitochondria. Therefore, the protein is candidate to be an essential component in the machinery for the mitochondrial import of PS. PMID- 9350037 TI - Inhibition of proliferation of T47D human breast cancer cells: alterations in progesterone receptor and p53 tumor suppressor protein. AB - We have investigated the influence of three structurally different but functionally related compounds [1, 10 ortho-phenanthroline (phenanthroline), Rifampicin and aurin tricarboxylic acid (ATA)] on the rate and the extent of proliferation of progesterone-responsive T47D human breast cancer cells. These compounds have previously been used in this laboratory and have been shown to modulate properties of nucleic acid binding proteins. Because p53 and the progesterone receptor (PR) are both DNA binding proteins that appear to regulate proliferation of breast cells, alterations in T47D cell p53 and PR levels were examined to determine their relevance in cell proliferation. T47D cells were grown in the absence of phenol red and in the presence of 5% fetal calf serum with or without charcoal stripping in the presence of the inhibitors. The rate of proliferation of cells grown in Rifampicin containing medium exhibited nearly 70% inhibition. Phenanthroline, a known metal chelator, was an effective inhibitor of proliferation at 3 mM reducing the cell number by more than 75%. ATA (0.24-2.4 micrograms/ml) inhibited the growth of the cells by nearly 50%. Analysis of the mechanism of action of these compounds revealed that treatment with these compounds caused specific changes in the molecular composition of T47D cell PR. Whereas ATA caused increased stability of PR isoforms, Rifampicin induced a upshift in the mobility of PR in SDS gels-a phenomenon associated with hyperphosphorylation of steroid receptors (SRs). Phenanthroline treatment (> 2 mM) caused a complete down-regulation of PR and the tumor suppressor protein, p53. The downregulation of p53 paralleled the changes in the molecular composition of PR. We propose that the inhibition of T47D cell proliferation by phenanthroline, Rifampicin and ATA results from a number of cellular changes that include regulation of p53 and PR. PMID- 9350038 TI - EPR detection of endogenous nitric oxide in postischemic heart using lipid and aqueous-soluble dithiocarbamate-iron complexes. AB - Spin-trapping techniques combined with electron paramagnetic resonance (EPR) spectroscopy to measure nitric oxide (NO) production were compared in the ischemic-reperfused myocardium for the first time, using both aqueous-soluble and lipophilic complexes of reduced iron (Fe) with dithiocarbamate derivatives. The aqueous-soluble complex of Fe and N-methyl-D-glucamine dithiocarbamate (MGD) formed MGD2-Fe-NO complex with a characteristic triplet EPR signal (aN 12.5 G and giso = 2.04) at room temperature, in native isolated rat hearts following 40 min global ischemia and 15 min reperfusion. Diethyldithiocarbamate (DETC) and Fe formed in ischemic-reperfused myocardium the lipophilic DETC2-Fe-NO complex exhibiting an EPR signal (g perpendicular = 2.04 and g parallel = 2.02 at 77 K) with a triplet hyperfine structure at g perpendicular. Dithiocarbamate-Fe-NO complexes detected by both trapping agents were abolished by the .NO synthase inhibitor, NG-nitro-L-arginine methyl ester. Quantitatively, both trapping procedures provided similar values for tissue .NO production, which were observed primarily during ischemia. Postischemic hemodynamic recovery of the heart was not affected by the trapping procedure. PMID- 9350039 TI - Phosphorylation of inhibitory subunit of troponin and phospholamban in rat cardiomyocytes: modulation by exposure of cardiomyocytes to hydroxyl radicals and sulfhydryl group reagents. AB - Myocytes were isolated from rat heart ventricles and then incubated with [32P] sodium phosphate to label intracellular ATP stores. Incubations of the [32P] labelled cardiomyocytes with a beta-adrenoceptor agonist isoproterenol (10 microM) and with a plant diterpene forskolin (100 microM) which directly stimulates adenylyl cyclase increased the phosphorylation of an inhibitory subunit of troponin (TN-I) and phospholamban (PLN). Brief exposure (1 min) of labelled myocytes to the hydroxyl radical generating system (H2O2 plus FeCl2) decreased markedly the stimulatory action of isoproterenol and forskolin on TN-I and PLN phosphorylation. Similar exposure of myocytes to 5-5'-dithiobis nitrobenzoic acid (DTNB) a sulfhydryl oxidizing reagent exerted little inhibitory effect on the isoproterenol or forskolin stimulated TN-I and PLN phosphorylation. In contrast exposure of myocytes to low concentrations (< 50 microM) of N ethylmaleimide (NEM) a sulfhydryl alkylating reagent augmented the stimulatory effect of isoproterenol on TN-I and PLN phosphorylation. The results further showed that brief treatment of myocytes to H2O2 plus FeCl2 markedly decreased isoproterenol-, but not forskolin-, stimulated cyclic AMP accumulation in the myocytes. The stimulatory action of NEM on the isoproterenol-stimulated TN-I and PLN phosphorylation appeared related to greater increase in the isoproterenol stimulated cyclic AMP accumulation in the NEM-treated cardiomyocytes. The results are consistent with the postulate that hydroxyl radical exposure of cardiomyocytes blunts the beta-adrenoceptor-mediated stimulation of adenylyl cyclase leading to decreased phosphorylation of TN-I and PLN and imply that such alterations account in part the reported depressed rate of relaxation of the myocardium exposed to oxygen free radicals. PMID- 9350040 TI - Protein phosphorylation in rat cardiac microsomes: effects of inhibitors of protein kinase A and of phosphatases. AB - The phosphorylation of rat cardiac microsomal proteins was investigated with special attention to the effects of okadaic acid (an inhibitor of protein phosphatases), inhibitor 2 of protein phosphatase 1 and inhibitor of cyclic AMP dependent protein kinase (protein kinase A). The results showed that okadaic acid (5 microM) modestly but reproducibly augmented the protein kinase A-catalyzed phospholamban (PLN) phosphorylation, although exerted little effect on the calcium/calmodulin kinase-catalyzed PLN phosphorylation. Microsomes contained three other substrates (M(r) 23, 19 and 17 kDa) that were phosphorylated by protein kinase A but not by calcium/calmodulin kinase. The protein kinase A catalyzed phosphorylation of these three substrates was markedly (2-3 fold) increased by 5 microM okadaic acid. Calmodulin was found to antagonize the action of okadaic acid on such phosphorylation. Protein kinase A inhibitor was found to decrease the protein kinase A-catalyzed phosphorylation of microsomal polypeptides. Unexpectedly, inhibitor 2 was also found to markedly decrease protein kinase A-catalyzed phosphorylation of phospholamban as well these other microsomal substrates. These results are consistent with the views that protein phosphatase 1 is capable of dephosphorylating membrane-associated phospholamban when it is phosphorylated by protein kinase A, but not by calcium/calmodulin kinase, and that under certain conditions, calcium/calmodulin-stimulated protein phosphatase (protein phosphatase 2B) is also able to dephosphorylate PLN phosphorylated by protein kinase A. Additionally, the observations show that protein phosphatase 1 is extremely active against the three protein kinase A substrates (M(r) 23, 19 and 17 kDa) that were present in the isolated microsomes and whose state of phosphorylation was particularly affected in the presence of dimethylsulfoxide. Protein phosphatase 2B is also capable of dephosphorylating these three substrates. PMID- 9350042 TI - A microassay for the detection of low levels of cytochrome P450 O-deethylation activities with alkoxyresorufin substrates. AB - A microfluorometric method for the detection of low levels of cytochrome P450 was developed to increase the sensitivity of the assay, since a low level of CYP450 associated enzymatic activities was expected in human placenta tissues and small samples of placenta (approximately 10 g) could be easily collected, stored and processed. The dual fluorescence assay of Kennedy et al. [1], which was developed to simultaneously quantitate microsomal proteins and ethoxyresorufin-O-deethylase (EROD) activity was adapted for 96 wells microtiter plates. Placental microsomes samples were analyzed. For samples obtained from non-smoking mothers from the general southern Quebec population, results ranged from less than 1-3.3 pmol/mg protein.min. Samples collected from smoking mothers showed activity levels ranging from 30-69 pmol/mg protein.min. These results showed the suitability of the microassay for measuring low level of CYP450 activity in tissues such as placenta. PMID- 9350041 TI - Impaired phosphatidylcholine biosynthesis and ascorbic acid depletion in lung during lipopolysaccharide-induced endotoxaemia in guinea pigs. AB - Injection of guinea pigs with a single dose of Escherichia coli lipopolysaccharide (3.2 mg/100 g) induces a reversible endotoxic shock that was evaluated by measuring plasma glucose levels and aspartate aminotransferase activity at 24 h after lipopolysaccharide injection. The hypoglycaemia and the increase in plasma aminotransferase activity observed, correlated with the alterations found during the recovery phase of endotoxic shock. When lipid peroxidation and some antioxidant systems were measured in lungs from treated animals, we only found differences in ascorbic acid content, that was decreased by 50%. Lipopolysaccharide treatment results in a depression of pulmonary phosphatidylcholine synthesis, that correlates with the surfactant deficiencies associated with respiratory illnesses in septic shock. Guinea pigs fed on a diet with a low content in ascorbic acid were more sensitive to endotoxin. In these animals we found no detectable levels of ascorbic acid in lung, whereas both vitamin E lung levels and pulmonary phosphatidylcholine synthesis were significantly decreased. Our results point out the significance of ascorbic acid in the protection against oxidative lung injury associated to endotoxaemia, and validate our shock model for further studies on the mechanisms of this pathological condition. PMID- 9350043 TI - Subcellular distribution of hexokinase isoenzymes in pancreatic islet cells exposed to digitonin after incubation at a low or high concentration of D glucose. AB - It was recently proposed that stimulation of pancreatic islet by D-glucose results in the translocation of glucokinase from the perinuclear area to the cell periphery, where the enzyme might conceivably interact with either the glucose transporter GLUT-2 or some other proteins and, by doing so, become better able to express its full catalytic activity. To explore the possible interaction between glucokinase and the cell boundary, dispersed rat pancreatic islet cells were preincubated for 60 min at a low (2.8 mM) or high (16.7 mM) concentration of D glucose, then exposed for 1 min to digitonin (0.5 mg/ml) and eventually centrifuged through a layer of oil for separation of the cell pellet from the supernatant fraction containing the material released by digitonin. Under these conditions, the bulk of lactate dehydrogenase and glutamate dehydrogenase activities were recovered in the supernatant fraction and cell pellet, respectively. The measurement of hexokinase isoenzyme activities in the two subcellular fractions, as conducted at low or high hexose concentrations and in either the absence or presence of exogenous hexose phosphates (3.0 mM glucose 6 phosphate and 1.0 mM fructose 1-phosphate) indicated a preferential location of the low-Km hexokinase in the cell pellet and of the high-Km glucokinase in the cytosolic fraction. Such a distribution pattern failed to be significantly affected by the concentration of D-glucose used during the initial incubation of the dispersed islet cells. These findings argue against the view that the glucose induced translocation of glucokinase would result in any sizeable binding of the enzyme to a plasma membrane-associated protein. PMID- 9350044 TI - Pb-induced alterations in tyrosine hydroxylase activity in rat brain. AB - Our previous studies have shown that exposure to low levels of Pb results in significant reductions in dopamine (DA) and its metabolites (3,4 dihyroxyphenylacetic acid, DOPAC and homovanillic acid, HVA) in nucleus acumbens (NA). This area of brain receives dopaminergic projections from the ventral tegmentum and is considered vital in manifestation of many behavioral responses. Similarly, basal and K(+)-induced release of DA was found significantly reduced in the Pb-exposed rats as compared to the controls in this brain region. Additional studies indicated that acute infusion of Pb in nucleus acumbens caused significant release of DA. Based on these observations it was postulated that the reductions in DA contents and in the basal and stimulus-induced release of DA in NA were manifestations of attenuated dopaminergic activity in this brain region. However, the mechanism of this attenuation is not yet clear. Studies reported here were designed to evaluate the role of a key regulatory enzyme in biosynthesis of DA, i.e. tyrosine hydroxylase (TH) in Pb-induced reductions in dopaminergic activity. The results of these studies indicated that 50 and 500 ppm Pb produced 22.8 and 56% inhibition of TH activity in vitro respectively, and that the enzyme activity was reduced to 43% in rats exposed to 50 ppm lead for 30 days as compared to the controls. The alterations in TH activity in Pb-exposed animals were further confirmed by Western blot analysis. Collectively, these results suggest that Pb-induced inhibition of TH activity in rat brain may contribute to the reductions in dopaminergic activity observed in Pb-exposed animals. PMID- 9350045 TI - Histidine and histamine metabolism in rat enterocytes. AB - To study the metabolic fate of L-histidine and histamine in rat isolated enterocytes, enterocytes were incubated in the presence of 0.1 mM L-[U-14C] histidine. At the rate of 11.1 +/- 2.7 pmol/10(6) cells/30 min, the amino acid was incorporated into cellular proteins. 80 microM cycloheximide, i.e. a protein synthesis inhibitor, inhibited this incorporation by 70 +/- 17%. L-histidine was used for cellular protein synthesis which depended on time and concentration. 0.1 mM L-[U-14C] histidine was little oxidized by intestinal cells, i.e. 0.12 +/- 0.06 pmol/10(6) cells/30 min, and was not converted into histamine. When 10 mM histamine was added to the incubation medium, it completely inhibited the incorporation of 0.1 mM [1,4-14C] putrescine into isolated enterocytes. In enterocyte homogenates, this corresponded to inhibition by histamine of putrescine incorporation as catalyzed by transglutaminase activity. Since histamine incorporation into TCA-precipitable material derived from enterocyte homogenates depended on time and concentration, we concluded that exogenous, but not de novo-formed histamine was able to compete with putrescine incorporation into enterocytes as catalyzed by transglutaminase activity. PMID- 9350046 TI - Lipogenesis in rat tissues following carbohydrate refeeding: spleen lipogenesis is modulated by insulin. AB - Intraperitoneal administration of [1,2-14C]-acetate to Wistar rats was used to assess tissue lipogenic rates after estimating the incorporation of the label into the tissular lipid fractions. Refeeding the animals with glucose (after an overnight fast) induced an increase in white adipose tissue (4.5 fold), liver (4.1 fold), small intestine (1.9 fold), carcass (2.9 fold) and spleen (3.7 fold) lipogenesis (expressed as the radioactivity present in the lipid fraction corrected by the plasma circulating radioactivity). No changes were found following refeeding in either brain or brown adipose tissue. Administration of mannoheptulose (an inhibitor of insulin secretion) to refed rats completely abolished the increased lipogenesis in white adipose tissue, liver, carcass, spleen and small intestine, thus suggesting that insulin secretion is involved in this phenomenon. This is the first report showing that spleen lipogenesis may be modulated by refeeding via insulin secretion and suggests an important role of this organ on the in vivo lipogenic response of the organism after carbohydrate refeeding. PMID- 9350047 TI - Trimetazidine increases phospholipid turnover in ventricular myocyte. AB - Trimetazidine (TMZ) is an anti-ischemic compound devoid of hemodynamic effects. It was recently suggested to induce cardiomyocyte protection by a mechanism involving lipid metabolism. The effects of TMZ were evaluated in rats on cardiac lipid composition, and in cultured rat cardiomyocytes on phospholipid metabolism. Rats were treated with TMZ for 4 weeks, and the fatty acid compositions were determined. Treatment with TMZ induced a significant decrease in phospholipid linoleic acid, balanced by a small increase in oleic and stearic acids. These changes were not correlated to alterations in plasma fatty acid composition. Cultured ventricular myocytes were treated with TMZ, 16 and 1 before experimentation. The time-dependent incorporation of radio labelled precursors of membrane phospholipids (3-inositol, 14C-ethanolamine, 14C-choline, 14C arachidonic acid, 10 mumol/L) was investigated. The cells were harvested 30, 60, 105 or 150 min after precursor addition. In TMZ-cells, arachidonic acid (AA) incorporation was increased in the phospholipids, but not in other lipid fractions. This increase elicited a net increase in the total AA uptake. The incorporation of 3-inositol in the phospholipids was strongly stimulated by TMZ, although the uptake of inositol was not altered. The difference was significant within 30 min, and after 150 min the phospholipid labelling in TMZ cells was higher by 70%. A similar result was obtained with ethanolamine as precursor, which turnover increased by 50% in TMZ-treated cells. Conversely, the incorporation of choline was not significantly affected by the presence of TMZ. In conclusion TMZ appears to interfere with the metabolism of phospholipids in cardiac myocytes in a manner which could indicate an increase of membrane phospholipid turnover. PMID- 9350048 TI - Expression of calcium-binding protein regucalcin mRNA in the cloned human hepatoma cells (HepG2): stimulation by insulin. AB - The expression of hepatic Ca(2+)-binding protein regucalcin in the cloned human hepatoma cells (HepG2) was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using rat liver regucalcin complementary DNA (0.9 kb of open reading frame). Regucalcin mRNA was expressed in HepG2 cells, although the mRNA was markedly expressed in normal rat liver. Moreover, regucalcin protein in HepG2 cells was detected by Western blot analysis using a polyclonal rabbit anti-regucalcin antibody. Regucalcin mRNA expression in HepG2 cells was clearly stimulated by the culture with insulin (10(-8) M) of the effective concentration. Regucalcin protein in HepG2 cells was also increased by the treatment of insulin (10(-8) M). The present results demonstrate that regucalcin is expressed in the transformed HepG2 cells, and that the expression is stimulated by insulin. PMID- 9350049 TI - Fate of Mycobacterium tuberculosis inside rat peritoneal macrophages in vitro. AB - Rat peritoneal macrophages in vitro were infected with Mycobacterium tuberculosis and the fate of M. tuberculosis inside macrophages was monitored. Alteration in the levels of nitric oxide (NO) measured in terms of nitrite formed, hydrogen peroxide (H2O2) and lysosomal enzymes such as acid phosphatase, cathepsin-D and beta-glucuronidase in macrophages following M. tuberculosis infection was also studied. Elevation in the levels of nitrite were observed from 72 h of M. tuberculosis infection. Irrespective of the time point, M. tuberculosis infected macrophages produced elevated levels of H2O2. Maximum increase in the level of acid phosphatase was observed from 72 h of M. tuberculosis infection, whereas maximum elevation in the level of beta-glucuronidase was observed 48 h after M. tuberculosis infection. However these microbicidal agents did not alter the intracellular viability of M. tuberculosis. PMID- 9350050 TI - Proteolysis activated protein kinase in Dictyostelium discoideum. AB - In the search for MBP phosphorylating activities in Dictyostelium discoideum, we have found a proteolysis-activated protein kinase. This activity which is distributed between the soluble and the particulate fractions of the cell, uses MBP and histone as substrate and has a molecular mass of 140 kDa as detected in an 'in situ' assay. This protein kinase has several features shared by the protein kinase C family, such as substrate specificity and sensitivity to proteolysis, but its molecular mass is much larger than that described for the known protein kinase C isoforms. To better characterize this activity we have studied its sensitivity to several protein kinase C inhibitors and activators. This protein kinase is activated neither by phorbol ester nor by phosphatidylserine or Ca2+. The activity is inhibited by staurosporine and PKC zeta pseudosubstrate, but is not affected by the specific protein kinase C inhibitor bisindolylmaleimide. These data lead us to propose that proteolytically activated Dictyostelium protein kinase belongs to the recently described protein kinase C-related family. PMID- 9350051 TI - Uptake and metabolism of lipoprotein-X in mesangial cells. AB - Progressive glomerulosclerosis is a major complication in patients with familial lecithin:cholesterol acyltransferase (LCAT) deficiency. The lack of LCAT activity results in the accumulation of an abnormal lipoprotein, lipoprotein-X (Lp-X), in the plasma of these patients. Lipoprotein-X contains high levels of unesterified cholesterol and phosphatidylcholine. Lp-X may play a role in the accumulation of lipids in the kidney, which in turn may lead to glomerulosclerosis. The objective of this study is to examine the uptake and metabolism of Lp-X by rat mesangial cells. Our results suggest that Lp-X is taken up by mesangial cells and that the lipids in Lp-X are metabolized. Lysosomes containing unesterified cholesterol and phosphatidylcholine, in a molar ratio similar to Lp-X, were synthesized to investigate the roles individual apolipoproteins (apo CI, II, III and E) play in the uptake of Lp-X. Both apo CI and CIII inhibited its uptake while apo CII (1.5 fold) and E (4 fold) stimulated the uptake of Lp-X. Very low density lipoprotein (VLDL) and low density lipoprotein (LDL) inhibited Lp-X uptake by mesangial cells. However, at higher concentrations of high density lipoprotein (HDL), the uptake of Lp-X was stimulated. Proteoglycans have an important role in regulating the uptake of Lp-X, while cytoskeleton-dependent phagocytosis and the scavenger receptor do not appear to be involved. PMID- 9350052 TI - Expression of the placenta-specific, 100 kDa ras GTPase activating protein in several human cancer cell lines and normal human tissues. AB - The ras GTPase activating protein (ras GAP), a regulator of Ras activity, has two isoforms; ras GAP 120 and ras GAP 100. The latter, whose molecular size is about 100 kDa, is generated alternative splicing from the ras GAP 120 gene and is considered placenta-specific, while the former is expressed ubiquitously. As point mutations of ras are frequently observed in human tumors, we investigated the expression of ras GAP in several human cancer cell lines and samples of human colon cancer using immunoprecipitation and immunoblot analysis with an anti-GAP monoclonal antibody, B4F8, as well as reverse transcription-polymerase chain reaction (RT-PCR). ras GAP 100 protein was detected in 4 of 9 colonic, 1 of 6 gastric and 1 of 4 lung cancer cell lines as well as ras GAP 120, but not in colon cancer specimens. In contrast, ras GAP 100 mRNA was present in all tested cell lines and colon cancer specimens. Then, we investigated ras GAP 100 expression in normal tissues, ras GAP 100 protein was not detected in human normal tissues except placenta. Contrary, ras GAP 100 message was expressed in normal tissues derived from liver, stomach, colon and lymphocyte although the level of which was smaller than that in placenta. These findings demonstrate that ras GAP 100, reportedly placenta-specific, is distributed in other normal tissues at least at mRNA level and its expression is augmented in some cancer cell lines. PMID- 9350053 TI - Characterization of ligand binding, DNA binding and phosphorylation of progesterone receptor by two novel progesterone receptor antagonist ligands. AB - In order to gain a better understanding of the distinctive mechanisms of the various types of antiprogestins, we have characterized in vitro ligand binding, specific DNA binding and phosphorylation of progesterone receptor (PR) from T47D cells after treatment of cells with progestins (progesterone, R5020) and antiprogestins (RU486, ZK98299, Org 31806 and Org 31710). Treatment of the cells with R5020 or PR antagonists, with the exception of ZK98299, resulted in a quantitative upshift of PR-A and PR-B indicative of ligand/DNA-induced phosphorylation of PR. Treatment of cells with RU486, Org 31710 or Org 31806, but not R5020 or ZK98299 resulted in detectable PR-progesterone response element complexes (PR-PREc) as assessed by gel mobility shift assay. Although treatment of cells with ZK98299, a type I PR antagonist, did not induce phosphorylation, the antiprogestins, Org 31806 and Org 31710, in a manner identical to RU486, did. Our data suggest that Org 31806 and Org 31710 affect properties of PR from T47D cells that are similar to RU486. PMID- 9350054 TI - Influence of ryanodine on the mechanical restitution and on the post extrasystolic potentiation of the guinea-pig ventricular myocardium. AB - This paper records the results of an investigation into potentiation and staircase phenomena in rightventricular guinea-pig papillary muscles with particular reference to the sarcoplasmic Ca(2+)-channel. As a tool to isolate the second ('late', 'tonic') component of isoproterenol-induced biphasic contractions ryanodine was used. On the evidence at present available the monophasic ryanodine resistant component of the twitch represents that portion of the activator calcium which reaches the troponin C directly, that is, not taking the roundabout way through the intracellular storage structures. In order to avoid functional instabilities of the isolated muscle preparation a short-time double rest stimulation programme was used which combines a number of different tests and gives information on (1) the post-rest potentiation, (2) the post-extrasystolic potentiation, (3) the mechanical post-rest recovery, (4) the interval-strength relationship, and (5) the mechanical restitution. The results of the present work show that under the influence of ryanodine (1) the Bowditch staircase, a typical feature of normodynamic mammalian ventricular preparations as well as of hypodynamic frog heart preparations, does not exist, (2) the post-extrasystolic potentiation disappears, (3) the curve reflecting the mechanical restitution, under normal in vitro conditions a monotonically increasing function, becomes biphasic within the relative refractory period, (4) the conspicuous depression of the isometric post-rest contraction for long lasting pauses interrupting the regular pacing rhythm, a typical feature of isolated guinea-pig ventricular tissue, is clearly diminished, and (5) the characteristic curve, reflecting the potentiation of the post-extrasystolic post-rest contraction as a function of the delay time preceding the extrastimulus, becomes displaced to the premature interstimulus interval. The concept of an 'extended 2-calcium-store model' is supported by this work. PMID- 9350055 TI - Alterations in susceptibility to carbon tetrachloride toxicity and hepatic antioxidant/detoxification system in streptozotocin-induced short-term diabetic rats: effects of insulin and Schisandrin B treatment. AB - The streptozotocin-induced short-term (2 week) diabetic rats showed an increase in susceptibility to carbon tetrachloride (CCl4)-induced hepatocellular damage. This diabetes-induced change was associated with a marked impairment in the hepatic glutathione antioxidant/detoxification response to CCl4 challenge, as indicated by the abrogation of the increases in hepatic reduced glutathione (GSH) level, glucose-6-phosphate dehydrogenase and microsomal glutathione S transferases (GST) activities upon challenge with increasing doses of CCl4. While the hepatic GSH level was increased in diabetic rats, the hepatic mitochondrial GSH level and Se-glutathione peroxidase activity were significantly reduced. Insulin treatment could reverse most of the biochemical alterations induced by diabetes. Both insulin and schisandrin B (Sch B) pretreatments protected against the CCl4 hepatotoxicity in diabetic rats. The hepatoprotection was associated with improvement in hepatic glutathione redox status in both cytosolic and mitochondrial compartments, as well as the increases in hepatic ascorbic acid level and microsomal GST activity. The ensemble of results suggests that the diabetes-induced impairment in hepatic mitochondrial glutathione redox status may at least in part be attributed to the enhanced susceptibility to CCl4 hepatotoxicity. Sch B may be a useful hepatoprotective agent against xenobiotics induced toxicity under the diabetic conditions. PMID- 9350056 TI - Cell-cycle regulated expression and serine phosphorylation of the myristylated protein kinase C substrate, SSeCKS: correlation with culture confluency, cell cycle phase and serum response. AB - We recently identified a novel myristylated protein kinase C (PKC) substrate, named SSeCKS (pronounced essex), whose transcription is suppressed > 15 fold in src- or ras-transformed rodent fibroblasts, but not in raf-transformed cells [1, 2]. SSeCKS associates with and controls the elaboration of a cortical cytoskeletal matrix in response to phorbol esters [2], and overexpression of SSeCKS causes growth arrest of untransformed NIH3T3 cells [3]. Our preliminary data suggested that SSeCKS functions as a negative mitogenic regulator by controlling cytoskeletal architecture and that serine phosphorylation of SSeCKS by kinases such as PKC alters its interaction with cytoskeletal matrices and its ability to control mitogenesis. Here, we determine the effects of culture confluency, growth arrest and serum response on the steady-state abundance of SSeCKS RNA and protein and on the relative level of phosphoserine-free SSeCKS. SSeCKS transcription is initially induced by serum factors and by contact inhibited growth rather than by cell-cycle arrest induced by serum starvation, hydroxyurea or nocodazole, and following serum-induced G1/S progression, SSeCKS transcription is suppressed. SSeCKS protein is hyperphosphorylated on serine residues during G1/S progression but not during the G2/M phase. Finally, we show that the induction of SSeCKS protein expression by contact inhibition is independent of SSeCKS' serum responsiveness. These data suggest that SSeCKS expression and function can be controlled at either the transcriptional or post translational level in response to serum factors and culture confluency. The data strengthen the notion that SSeCKS plays an important, yet transient, role in cell cycle progression from G0 to G1 that differs from its role in controlling contact inhibited growth. PMID- 9350057 TI - The integrity of the SH3 binding motif of AFAP-110 is required to facilitate tyrosine phosphorylation by, and stable complex formation with, Src. AB - The actin filament-associated protein AFAP-110 forms a stable complex with activated variants of Src in chick embryo fibroblast cells. Stable complex formation requires the integrity of the Src SH2 and SH3 domains. In addition, AFAP-110 encodes two adjacent SH3 binding motifs and six candidate SH2 binding motifs. These data indicate that both SH2 and SH3 domains may work cooperatively to facilitate Src/AFAP-110 stable complex formation. As a test for this hypothesis, we sought to understand whether one or both SH3 binding motifs in AFAP-110 modulate interactions with the Src SH3 domain and if this interaction was required to present AFAP-110 for tyrosine phosphorylation by, and stable complex formation with, Src. A proline to alanine site-directed mutation in the amino terminal SH3 binding motif (SH3bm I) was sufficient to abrogate absorption of AFAP-110 with GST-SH3STC. Co-expression of activated Src (pp60(527F)) with AFAP-110 in Cos-1 cells permit tyrosine phosphorylation of AFAP-110 and stable complex formation with pp60(527F). However, co-expression of the SH3 null-binding mutant (AFAP71A) with pp60(527F) revealed a 2.7 fold decrease in steady-state levels of tyrosine phosphorylation, compared to AFAP-110. Although a lower but detectable level of AFAP71A was phosphorylated on tyrosine, AFAP71A could not be detected in stable complex with pp60(527F), unlike AFAP-110. These data indicate that SH3 interactions facilitate presentation of AFAP-110 for tyrosine phosphorylation and are also required for stable complex formation with pp60(527F). PMID- 9350058 TI - Influence of phospholipid long chain polyunsaturated fatty acid composition on neonatal rat cardiomyocyte function in physiological conditions and during glucose-free hypoxia-reoxygenation. AB - There is evidence that dietary polyunsaturated fatty acids (PUFA) may protect against cardiovascular diseases, but the involvement of the cardiac muscle cell in this beneficial action remain largely unknown. The present study compared the respective influence of n-3 and n-6 PUFA on the function of cultured neonatal rat cardiomyocytes (CM). Cells were grown for 4 days in media enriched either n-3 (eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA) or n-6 (arachidonic acid, AA) PUFA. The PUFA n-6/n-3 ratio in the phospholipids was close to 1 and 20 in the n-3 and n-6 cells, respectively. The transmembrane potentials were recorded using microelectrodes and the contractions were monitored with a photoelectric device. In physiological conditions, the increase of n-6 PUFA level in the phospholipids resulted in a significant decrease in the maximal rate of initial depolarization (-16%). In opposition, the action potential amplitude and duration were not altered, and the cell contraction outline was not affected. Ischemia was simulated in vitro using a substrate-free, hypoxia-reoxygenation procedure in a specially designed gas-flow chamber. The progressive loss of electrical activity induced by the substrate-free, hypoxic treatment was affected by the n-6/n-3 ratio, since the n-6 rich CM displayed a slower depression of the AP amplitude and duration parameters. Conversely, the recovery of the resting potential (MDP) during reoxygenation was faster in n-3 CM, whereas the recovery of the contraction parameters was unaffected by the fatty acid composition of the cells. These results suggested that, in physiological conditions, the modification of long chain PUFA balance in the phospholipids of cardiac muscle cells may modulate the initial AP upstroke, which is governed by sodium channels. Moreover, the presence of n-3 PUFA appeared to accelerate the electrical depression during substrate-free hypoxia but in turn to allow a faster recovery upon reoxygenation. PMID- 9350059 TI - Glucose-induced positive cooperativity of fructose phosphorylation by human B cell glucokinase. AB - Human B-cell glucokinase displays sigmoidal kinetics towards D-glucose or D mannose, but fails to do so towards D-fructose. Yet, D-glucose, D-mannose and 2 deoxy-D-glucose confer to the enzyme positive cooperativity towards D-fructose. For instance, in the presence of 5 mM D-[U-14C]fructose, its rate of phosphorylation is increased to 214.3 +/- 11.0%, 134.0 +/- 4.3% and 116.5 +/- 3.0% of paired control value by D-glucose, D-mannose and 2-deoxy-D-glucose (each 6 mM), respectively. D-glucose and, to a lesser extent, D-mannose also display reciprocal kinetic cooperativity. D-fructose, however, fails to affect D-glucose or D-mannose phosphorylation under conditions in which positive cooperativity is otherwise observed. These findings are relevant to the reciprocal effects of distinct hexoses upon their phosphorylation by B-cell glucokinase and, as such, to the metabolic and functional response evoked in pancreatic islet B-cells by these sugars, when tested either separately or in combination. PMID- 9350060 TI - Disappearance of periodic sharp wave complexes in Creutzfeldt-Jakob disease. AB - Periodic sharp wave complexes (PSWC) are sensitive and specific of Creutzfeldt Jakob disease (CJD). Once they have emerged, PSWC may exceptionally disappear in the terminal stage of the disease, as a consequence of the flattening of scalp electroencephalogram (EEG). We document the disappearance of PSWC in serial EEG during the clinical course in two women (57 and 70 years of age) with pathologically proven CJD. Despite PSWC disappearance, diffuse theta-delta activity was still well recognizable. Moreover, external stimuli failed to trigger PSWC. The absence of PSWC in CJD might be due to the timing and frequency of EEG recordings. PSWC disappearance should not be interpretated as evidence against the diagnosis of CJD. PMID- 9350062 TI - Reproducibility of normal facial motor nerve conduction studies and their relevance in the electrophysiological assessment of peripheral facial paralysis. AB - To determine the intra-examiner intertrial reproducibility of normal facial motor nerve conduction studies (FNCS) and their relevance in electrophysiological assessments of peripheral facial paralysis, 52 patients with acute unilateral Bell's palsy were examined on two separate occasions 1 months apart. Three electroneurographic methods were assessed. On the unaffected side of the face, FNCS are reliable when performed by a single examiner over time. Nevertheless, compound muscle action potential (CMAP) baseline-to-peak and peak-to-peak amplitude showed a rather high intertrial variability. Reproducibility of the assessed surface electrode recording procedures was similar. Regarding the affected side, in patients with mild axonotmesis of the facial nerve variations of electroneurographic parameters 1 months apart fell within the range of normal intertrial variability. In patients with severe or moderate axonotmesis, the distal latency and the M wave amplitude variations showed significant intertrial variations. Reproducibility of FNCS appears to be similar to that found in limb motor nerves. Normal variability curtails the sensitivity of FNCS in detecting mild facial nerve axonotmesis, although this technique remains useful in severe cases. PMID- 9350061 TI - Diagnostic value of multimodal evoked potentials in HIV-1 infected children. AB - Evoked potentials (EP) help guide the diagnosis of central nervous system involvement in demyelinating pathologies regarding both children and adults, and in human immunodeficiency virus-1 (HIV-1) correlated pathologies only in regard to adult patients. EP have been shown to be useful in highlighting early signs of the disease. We therefore studied EP in HIV-1 infected children with the aim of verifying the association of results with disease progression, clinical signs and electroencephalogram, and individualizing the most reliable test. Thirty-six patients (20 male and 16 female subjects, age range: 10 months to 12 years) belonging to a group of 45 symptomatic subjects seen at the Pediatric Department were included into the study from November 1991 to December 1994. Ten presented with neurological signs as of disease onset, eight others developed encephalopathy during the follow-up. One hundred seventeen EP, i.e., 27 pattern visual, 64 flash visual and 26 brain stem auditory EP, were recorded. Univariate statistical analysis using the Wilcoxon-Mann-Whitney U test and Student's t test was done. As a whole, we found 22.5% of abnormal EP in subjects without neurological signs and 28.3% in subjects with neurological signs. Results that were obtained suggested a close relationship between both the pattern of visual and brain stem auditory EP exams and disease progression. PMID- 9350063 TI - Coactivation of the ulnar nerve in motor tests for carpal tunnel syndrome. AB - Coactivation of the ulnar nerve at the wrist can be a source of error in tests for carpal tunnel syndrome (CTS). We compared the effects of coactivation in two tests for CTS: the abductor pollicis brevis-distal motor latency (APB-DML) and lumbrical-interosseus-distal motor latency difference (2LI-DML). We studied 33 hands of 25 consecutive patients referred for suspected CTS. In severe CTS, when selective supramaximal stimulation of the median nerve was impossible, all APB compound muscle action potentials (CMAP) showed abnormalities, indicating volume conduction of ulnar muscle activation. 2LI-DML in these hands led to falsely normal test results, as two identical CMAP were obtained after median and ulnar stimulation. In less severe CTS, warning signs indicating coactivation were observed in APB-DML virtually as often as in 2LI-DML. Undetected coactivation was more likely to be associated with false normal test results in 2LI-DML than in APB-DML. PMID- 9350065 TI - Dissociation between water and lithium transport during acute changes in plasma potassium concentration in dog kidney. AB - Lithium clearance is often used as a marker for proximal tubular water transport. Proximal tubular transport may be modulated by changing plasma potassium concentration. The aim of the present study was to examine the effect of acute changes in plasma potassium concentration on proximal tubular fluid and lithium transport. Clearance studies were performed in seven anaesthetised, volume expanded dogs treated with amiloride (1 mg kg-1 body weight) to block distal tubular potassium secretion, and with bumetanide (30 micrograms kg-1 body weight) to inhibit sodium reabsorption in Henle's loop. When plasma potassium concentration was raised from 2.6 +/- 0.2 to 7.9 +/- 0.2 mmol l-1, water reabsorption decreased from 23.9 +/- 2.9 to 19.8 +/- 2.2 ml min-1, whereas lithium reabsorption increased from 10.5 +/- 2.3 to 18.1 +/- 2.3 mumol min-1, at constant glomerular filtration rate. We conclude that acute elevation of plasma potassium concentration inhibits proximal tubular fluid reabsorption, but stimulates renal lithium reabsorption. Thus, lithium reabsorption cannot be used as a marker for proximal tubular transport during acute changes in plasma potassium concentration. PMID- 9350064 TI - Reference intervals for the glomerular filtration rate and cell-proliferation markers: serum cystatin C and serum beta 2-microglobulin/cystatin C-ratio. AB - Recent studies have indicated that serum and plasma cystatin C are better markers for glomerular filtration rate (GFR) than serum creatinine, ubiquitously used for this purpose. To fully exploit the value of serum and plasma cystatin C as GFR markers, reliable age and sex-correlated reference intervals are required. The present study comprised cystatin C determinations in plasma and sera from 259 individuals from a well-defined area in the southernmost part of Sweden. From demographic lists two men and two women were randomly selected from each one-year birth cohort above 20 years of age. No sex differences were found for plasma and serum cystatin C, whereas an increase in the cystatin C levels with age was noted, corresponding to the known age-related decrease in GFR. The following reference intervals are recommended for practical clinical use: S-Cystatin C (both sexes): 20-50 years, 0.70-1.21 mg l-1 and 50+ years, 0.84-1.55 mg l-1. The same samples were also used for determination of beta 2-microglobulin levels in order to calculate reference intervals for the beta 2-microglobulin/cystatin C ratio, which is a more distinct marker for cell proliferation, particularly lymphoproliferation, than is the serum level of beta 2-microglobulin alone, since the ratio should be virtually uninfluenced by GFR. The beta 2 microglobulin/cystatin C-ratios were uninfluenced by sex and age and 1.45-2.43 is recommended as the serum reference interval for practical clinical use. Serum creatinine was determined in the same samples and the creatinine level was found to be strongly influenced by sex and weakly by age. PMID- 9350066 TI - Assessment of different markers of bone resorption in postmenopausal osteoporotic women treated with pamidronate. AB - The aim of this study was to evaluate the different bone resorption markers, total pyridinoline (Pyr) and total deoxypyridinoline (Dpyr), assessed by a HPLC method, free Dpyr, assessed by a new immunoassay, and urinary excretion of hydroxyproline (OH-proline), in postmenopausal osteoporotic women during long term treatment with pamidronate. A total of 60 postmenopausal women with previous distal forearm fracture were included in this 12-month placebo-controlled and double-blind study, where intermittent oral pamidronate, 75 or 150 mg, or placebo were given daily for 4 weeks, every 16 weeks. After 1 week a significant reduction in urinary excretion of total Dpyr was observed in the group treated with 150 mg pamidronate compared to the placebo (p < 0.01) and to the 75-mg group (p < 0.001). A maximal 50.4% decrease in total Dpyr, (p < 0.0001 compared to the placebo group, p < 0.01 compared to the 75-mg group), was observed after 3 weeks of treatment with 150 mg pamidronate, and this decrease persisted to week 52. After 4 weeks of treatment with 150 mg pamidronate the maximal decrease in free Dpyr was only 26.5%, which persisted during 12 months of treatment. Decreases in urinary excretion of total Pyr and OH-proline were less than the decreases in total Dpyr. The correlation between total Dpyr (HPLC method) and free Dpyr (Pyrilinks-D assay) at baseline was r = 0.91. Total Dpyr assessed by the HPLC method reflects the pamidronate-induced decrease in bone resorption, and the changes in this resorption marker were more pronounced than changes in free Dpyr, total Pyr and OH-proline. In this study free Dpyr analysis was less suitable for reflecting bone resorption during bisphosphonate therapy. PMID- 9350067 TI - Chronic erythropoietin treatment enhances endogenous nitric oxide production in rats. AB - To examine the effect of chronic administration of recombinant human erythropoietin (rHuEPO) on endogenous nitric oxide (NO) activity, we treated Sprague-Dawley rats with rHuEPO (100 IU kg-1 or 300 IU kg-1) or a corresponding vehicle for 2 weeks, administered subcutaneously on alternate days. Treatment elicited increases in haematocrit and systolic blood pressure in a dose-dependent fashion. Simultaneous administration of NG-nitro-L-arginine methyl ester (L-NAME, 20 mg dl-1 of drinking water), but not aminoguanidine (400 mg dl-1), induced a further significant rise in blood pressure. The effect of L-NAME was inhibited by a large dose of L-arginine (2.0 g dl-1). Polycythaemia and hypertension induced by chronic rHuEPO therapy were associated with increased urinary NO2- and NO3- (NOx-) excretion, while co-administration of L-NAME, but not aminoguanidine, reduced NOx- excretion. Our results indicate that chronic rHuEPO treatment has a significant pressor effect, but induces a compensatory increase in the steady state release of NO by constitutive NO synthase in normal rats. Such enhanced NO synthesis may act as a protective mechanism against the hypertensive effect of rHuEPO. PMID- 9350068 TI - n-3 fatty acids do not decrease plasma endothelin levels in healthy individuals. AB - The effect of three different doses of n-3 polyunsaturated fatty acids (PUFA) on endothelin-1 (ET-1) was studied. Study 1 included 40 healthy volunteers randomized to a single supplement of 20 g of n-3 or n-6 PUFA. Plasma ET-1 was measured 14 h after ingestion, and no changes in plasma ET-1 after intake of n-3 PUFA were observed, compared to baseline values. In study 2, 32 subjects had 0.65 g of n-3 PUFA or a fat mixture per day for 12 weeks. No changes in plasma ET-1 were found after the oil supplements. Finally, 22 persons had 4 g of n-3 PUFA for 6 weeks. A significant increase in plasma ET-1 was seen in this group after the supplement. Thus, n-3 PUFAs do not lower plasma levels of ET-1, the most potent vasoconstrictor known. PMID- 9350069 TI - Salt restriction: effects on lipids and insulin production in hypertensive patients. AB - The object of the study was to evaluate blood pressure, insulin and glucose metabolism, and serum lipids in hypertensive patients, during 8 weeks on a moderately salt-restricted diet. A double-blind, cross-over study was conducted with hypertensive patients following a moderately salt-restricted diet. Patients were randomised to sodium capsules in one period and placebo capsules during the other period. After a 1-month run-in period, 13 males and three females with mild to moderate essential hypertension (mean age 50 years) complied with a salt reduced diet. They were randomized to a salt-supplemented group (5 capsules of 10 mmol sodium per capsule) or a salt reduced diet group (5 capsules of placebo) with cross-over after 8 weeks. Serum insulin, insulin C-peptide, and glucose were measured, fasting and 30 min after a 75-g glucose load. Serum lipids and lipoproteins constituting an atherogenic index were measured, along with blood pressure and 24-h urine excretion of sodium and chloride. Non-significant reductions of systolic and diastolic blood pressure (4 mmHg, p = 0.06, and 2 mmHg, p = 0.13, respectively) were observed during the reduced-salt period. The changes observed for fasting insulin, insulin C-peptide, glucose, serum lipids and the atherogenic index were also non-significant. It is concluded that moderate salt restriction seems not to adversely influence insulin resistance or serum lipids in hypertensive patients. PMID- 9350070 TI - Effects of nandrolone decanoate (Decadurabolin) on serum Lp(a), lipids and lipoproteins in women with postmenopausal osteoporosis. AB - Although lipoprotein(a) (Lp(a)) concentrations are mainly regulated genetically, it has been reported that variations in sex hormone concentrations may have effects on serum Lp(a). We evaluated the effect of nandrolone decanoate, a testosterone-derived synthetic anabolic steroid, on serum Lp(a), lipids and lipoproteins in 19 postmenopausal women who were given parenteral nandrolone decanoate (Decadurabolin) once a week for 3 weeks. At the 4th week, a significant decrease was observed for total cholesterol (p = 0.003), Lp(a) (p = 0.0003), apolipoprotein A-I (apo A-I) (p < 0.0001), and high density lipoprotein cholesterol (HDL-C) (p < 0.0001). Moreover, a significant decrease in serum albumin concentration (p = 0.002) was concomitantly observed. We conclude that the administration of nandrolone decanoate significantly affects the lipid profile of postmenopausal women, showing controversial effects in terms of cardiovascular risk. PMID- 9350071 TI - Smoking cessation does not change urinary albumin excretion in normal subjects. AB - The aim of the study was to examine the effect of smoking cessation on urinary albumin excretion (UAE) in normal subjects. The study consisted of two parts. The first was a randomized 4-week study, in which 182 heavy smokers were asked to quit smoking immediately (n = 69, available for analysis) or to continue smoking for another 4 weeks (n = 70, available for analysis). After 4 weeks, the latter group was also asked to stop smoking. The second part was a non-randomized follow up study comparing UAE in 33 unsuccessful and 57 successful quitters followed for 26 weeks. Measurements of UAE (ELISA) were taken from 24-h urine samples before smoking cessation, after 4 weeks, and after 26 weeks. After 4 weeks, no statistically significant change in UAE was found within each group or between quitters and smokers. The 95% confidence intervals of the change in log UAE were 7.4 to 9.9% of the initial value in the smoker group and -4.9 to 11.3% in the quitter group. In the second part of the study, after 26 weeks, a 16% increase (95% confidence interval 5.5 to 26.5%) in mean log UAE was found in the group that had stopped smoking (p < 0.003), but no statistically significant difference in UAE between continued smokers and quitters was found after adjusting for the baseline level (ANCOVA). In conclusion, smoking cessation seems to have no effect on UAE within the physiological range in normal subjects over an observation period of 4 weeks, and no sign of a decrease in UAE was seen after 26 weeks of smoking cessation. PMID- 9350072 TI - Metformin increases total serum homocysteine levels in non-diabetic male patients with coronary heart disease. AB - It is known that the metabolism of homocysteine (Hcy) depends on the vitamins B6, B12 and folate, and furthermore that metformin reduces serum vitamin B12 levels. In order to investigate whether metformin treatment affects serum total Hcy (tHcy) levels we performed an open, prospective, randomised study in 60 non diabetic male patients with cardiovascular disease. After a 4-week run-in period with lovastatin 40 mg day-1, and diet and lifestyle advice, patients were randomised into two groups, both continuing the run-in treatment. One group received metformin up to 2000 mg day-1, whereas the control group got no additional treatment. After 12 and 40 weeks of metformin treatment, tHcy levels increased moderately but significantly by 7.2% (p < 0.05) and 13.8% (p < 0.05) in the metformin group relative to the control group, whereas serum vitamin B12 levels decreased by 13.4% (p < 0.0005) and 17.7% (p < 0.0005), respectively. Serum folate levels did not change after 12 weeks, but decreased by 8.0% after 40 weeks (p = 0.061) relative to the control group. Serum levels of total cysteine and methylmalonic acid (MMA) did not change. In conclusion, metformin treatment increased tHcy levels and decreased levels of vitamin B12 and folate. Since MMA levels were unchanged, it remains an open question whether the increase in tHcy levels is secondary to reduced vitamin B12 levels, folate levels or a combination of both. PMID- 9350073 TI - A new, fast and reliable radioimmunoassay of brain natriuretic peptide in human plasma. Reference values in healthy subjects and in patients with different diseases. AB - A new, fast and reliable radioimmunoassay for measurement of brain natriuretic peptide (BNP) in human plasma has been developed and its application is reported in healthy subjects and in patients with congestive heart failure, chronic renal failure, liver cirrhosis and essential hypertension. The antibody was raised in rabbits, the tracer was made by the iodogen method and polyethylene glycol was used for separation of free and bound tracer. BNP was extracted from plasma using Sep-Pak C18 cartridges. The recovery of unlabelled BNP added to plasma was 77.5 +/- 6.2% (mean +/- SD). The detection limit in plasma was 0.55 pmol l-1. No cross reactivity existed with the natriuretic peptides ANP, CNP or urodilatin. In 124 healthy subjects the mean BNP was 1.8 +/- 1.0 pmol l-1 (SD), range 0.6-5.5. BNP increased slightly with age, was higher in women than men and had no circadian rhythm. In eight patients with congestive heart failure the median BNP level was 30.5 pmol l-1, range 3.9-65.3. In 14 patients with chronic renal failure the median BNP level was 50.5 pmol l-1, range 10.9-219.8 before dialysis, and 38.0 pmol l-1, range 9.4-180.0 immediately following dialysis. In 25 patients with liver cirrhosis the median BNP value was 7.8 pmol l-1, range 1.2-43.1. There was no difference between patients with or without ascites. In 18 medically treated patients with essential hypertension the median BNP level was 5.0 pmol l-1, range 1.2-45.5 pmol l-1. PMID- 9350074 TI - Minimal residual disease in B-lymphoproliferative disorders by PCR detection of immunoglobulin heavy chain gene recombination. AB - We amplified and sequenced the rearranged immunoglobulin heavy chain VDJ genomic unit in B-leukemias and used it as a clone-specific marker for the molecular monitoring of the patients during and after therapeutic treatment. The method described is patient-specific rather than disorder-specific, more sensitive and less time-consuming than other conventional techniques for the detection of minimal residual disease. We propose reproducible and quick procedures, from DNA extraction to Southern blotting, that can be easily performed in any clinical laboratory and also applied to other kinds of investigation. PMID- 9350075 TI - An improved rapid troponin T test with a decreased detection limit: a multicentre study of the analytical and clinical performance in suspected myocardial damage. AB - In a multicentre study, we evaluated the analytical and diagnostic performance of the second version of the TROPT rapid test (TROPT 2, CARDIACT in the US). We tested TROPT 2 on 796 blood samples from 487 patients admitted on suspicion of myocardial infarction between 1 and 72 h after onset of symptoms and determined cTnT ELISA and CK MB mass in the corresponding serum samples. Frequency distributions of the results with TROPT 2 showed a detection limit of 0.18 microgram/l (for 50% positive results) as determined by the quantitative cTnT ELISA method. In a total of 796 samples the sensitivities in the detection of myocardial infarction (WHO criteria) 8-12 h after onset of symptoms were highest for cTnT ELISA (98%), followed by the rapid assay and CK MB mass (92%). A subgroup of 87 patients was primarily classified by the WHO criteria for definite infarction. Based on the maximum values within each patient time-series, diagnostic sensitivities for infarction were 100% for TROPT2, cTnT ELISA and CK MB mass. The corresponding specificities were 90%, 82% and 100%, respectively. After reclassification summarizing all cases of myocardial damage (acute and subacute myocardial infarctions and minor myocardial damage) the sensitivities were 87% (TROPT2), 100% (cTnT ELISA) and 71% (CK MB mass). The specificities of all three markers were 100%. Over 50% of all cases of minor myocardial damage were detected by TROPT 2. The clinical evaluation showed that the diagnostic performance of TROPT 2 is only slightly lower than that of cTnT ELISA. PMID- 9350076 TI - Comparison of gas chromatography and immunoassay methods for the detection of atrazine in water and soil. AB - Leachate and soil samples collected from different tillage systems were analyzed for atrazine using gas chromatography (GC) and an enzyme-linked immunosorbent assay (ELISA) based on magnetic particle technology. Solid-phase extraction (SPE) was used to concentrate atrazine residues in leachate samples and soil extracts before GC analysis. Atrazine concentrations determined by GC ranged from 0.1 to 600 micrograms L-1 for water samples and from 1.0 to 700 micrograms kg-1 for soil samples. Atrazine concentrations in 92 leachate samples as determined by ELISA were well-correlated (R = 0.97) with GC levels over the entire concentration range. Soil samples (215) were prepared and analyzed by three combinations of extraction/detection methods: 1) conventional extraction for GC/detection by GC analysis; 2)conventional extraction for GC/detection by ELISA analysis; 3)extraction for ELISA using a commercially available field kit/detection by ELISA analysis. Methanol (MeOH) in water was the common extractant. Although the initial comparison of soil extracts between the two different systems (Method 1 versus Method 3) was favorable (R = 0.97), two-thirds of the samples contained levels below the lower threshold for atrazine detection by both methods and some extracts were perceived to provide unfavorable substrate conditions (> 10% MeOH). Elimination of these data points reduced the correlation value (R = 0.77). To determine possible sources of variability, the extraction and detection methods were examined separately. In a comparison of extraction methods (Method 2 versus Method 3), ELISA analysis of kit extracts underestimated (R = 0.71) atrazine levels compared to those conventionally extracted, suggesting that differences in extraction time between methods may have accounted for reduced kit efficiency. Where detection methods (Method 1 versus Method 2) were compared on specific extracts (< 10% MeOH), good agreement (R = 0.99) was achieved between ELISA and GC values, illustrating that control of extractant concentration is critical in using this assay for atrazine detection in soil. PMID- 9350077 TI - Carbofuran degradation in soil profiles. AB - Two soils, Puyallup fine sandy loam from Puyallup, WA, and Ellzey fine sand from Hastings, FL, each with a prior history of carbofuran exposure but with different pedological and climatological characteristics, were found to exhibit enhanced degradation toward carbofuran in surface and subsurface soil layers. The treated Puyallup and Ellzey soils exhibited higher mineralization rates for both the carbonyl and the aromatic ring of carbofuran when compared to untreated soils. Disappearance rates of [14C-URL (uniformly ring labeled)] carbofuran in the treated Ellzey soil was faster than in untreated soil, and also faster in surface soil than in subsurface soil. Initial degradation patterns in the treated Ellzey soil were also different from those in the untreated soil. The treated Ellzey soil degraded carbofuran mainly through biological hydrolysis, while untreated soil degraded carbofuran through both oxidative and hydrolytic processes. PMID- 9350078 TI - Bioconcentration and excretion of fenitrothion in the brain of the eel (Anguilla anguilla). AB - The bioconcentration of fenitrothion in the brain of the european eel (Anguilla anguilla) and its posterior elimination have been studied. Animals were exposed to a sublethal concentration of fenitrothion (0.04 mg/L) for 96 hours in a flow through test system. After this pesticide exposure, animals were transferred to clean water for 72 hours more. Bioconcentration and elimination processes of fenitrothion were studied in blood and brain. This insecticide showed a strong tendency to bioconcentrate into selected tissues. A steady-state was observed in blood in few hours. Highest accumulation was detected in brain, where any steady state could be observed. Elimination started rapidly from both tissues when a recovery period was allowed. Elimination kinetics were adjusted to one compartment model. K2 of 0.015 and 0.044 hr-1 were calculated for fenitrothion in blood and brain. These K2 values were related with a relatively short half-live of fenitrothion in the analyzed tissues; probably due to the low biotransformation rate of this toxicant in the european eel. That fact would protect the animals against many biotransformation products even more toxic than the parent fenitrothion. PMID- 9350079 TI - Response and recovery of acethylcholinesterase activity in the European eel Anguilla anguilla exposed to fenitrothion. AB - European eel (Anguilla anguilla) were exposed to sublethal fenitrothion concentrations in a continuous flow-through system for 4 days. Muscle acetylcholinesterase (AChE) activity was evaluated after 2, 8, 12, 24, 32, 48, 56, 72 and 96 hours pesticide exposure. Results showed that AChE activity in eel muscle tissue decreased as concentration of fenitrothion increased. Pesticide induced significant inhibitory effects on the AChE activity of A. anguilla ranging from > 35% inhibition at sublethal concentration of 0.02 ppm to > 44% inhibition at sublethal concentration of 0.04 ppm. Eel were exposed to both fenitrothion concentrations for 96 hours and then allowed a period of recovery in pesticide-free water. Samples were taken out at 8, 12, 24, 48, 72, 96, 144 and 192 hours and eel muscle AChE activity was evaluated. Following 1 week of recovery, the AChE activity for those animals previously exposed to 0.02 and 0.04 ppm fenitrothion was still different from the controls. So, the AChE activity of eel muscle at the end of the recovery phases remained significantly depressed. PMID- 9350080 TI - Balancing phosphine in manure fermentation. AB - The evolution of phosphine gas during the anaerobic batch fermentation of fresh swine manure was detected and correlated to the production of methane and hydrogen sulphide. A close temporal relationship between phosphine liberation and methane formation was found. However, the gaseous phosphine released from manure during fermentation only represents a tiny fraction of the overall phosphine balance. The majority of phosphine is captured in solid manure constituents. This matrix-bound phosphine is eliminated by more than 50% during anaerobic batch fermentation. Seasonally determined phosphine concentrations in biogas and manure from two large-scale manure treatment plants also revealed net losses of phosphine in fermentation. Consequently, manure has to be considered more as a sink of phosphine rather than a phosphine-generating medium. Furthermore, a close relationship between phosphine in the feed of swine and manure of these swine was observed, implying that phosphine residues in the feed (possibly as a result of grain fumigation) represent an important source of phosphine in manure technologies that is relevant before the faecals of swine enter manure treatment plants. PMID- 9350084 TI - The management of urinary tract infection in children. PMID- 9350085 TI - Acamprosate for alcohol dependence? PMID- 9350086 TI - Humalog--a new insulin analogue. PMID- 9350087 TI - Stopping panic attacks. PMID- 9350088 TI - Progenitor cell transplantation. PMID- 9350090 TI - Value of prostatic cancer screening in patients treated for benign prostatic hyperplasia with medical or minimally invasive modalities. AB - OBJECTIVE: To assess the likelihood of overlooking the diagnosis of prostate cancer (PCA), using current screening methods, in patients treated for benign prostatic hyperplasia (BPH) with medical or minimally invasive treatment modalities, which do not produce tissue specimens for histologic review. To appraise this, we examined the impact of the preoperative use of prostatic specific antigen (PSA) in combination with transrectal ultrasound (TRUS) and systematic sextant prostate needle biopsy (PNbx) on the subsequent incidence of stage A PCA in patients undergoing transurethral resection of the prostate (TURP). METHODS/RESULTS: After excluding all patients found to have PCA during pretreatment screening, 485 patients who underwent TURP for presumed BPH from 1976 to 1994, were reviewed. From 1976 to 1989, PSA was not used for pretreatment screening, and stage A PCA was diagnosed in 11.4% of 317 patients. In 1990 and 1991, pretreatment screening included PNbx obtained under ultrasound guidance for PSA > or = 15.0 ng/ml. Stage A PCA was diagnosed in 14.2% of 63 patients. From 1992 to 1994, pretreatment screening included systematic sextant PNbx performed for PSA > 4.0 ng/ml, and stage A PCA was diagnosed in 2.8% of 105 patients. The difference in incidence of stage A PCA between the first two groups and group three was significant (p = 0.021); so is the difference in incidence of stage A2 (p = 0.037). For stage A1, the difference did not reach statistical significance (p = 0.089). CONCLUSION: Our findings suggest that systematic sextant PNbx for PSA > 4.0 ng/ml significantly reduces the risk of overlooking prostate cancer in patients undergoing treatment of BPH with modalities that do not provide tissue specimens. PMID- 9350092 TI - Legionnaires' disease in residents of Scotland: 1996. PMID- 9350091 TI - Legionnaires' disease in residents of England and Wales: 1996. AB - Two hundred and one cases of legionnaires' disease were reported to the PHLS Communicable Disease Surveillance Centre in 1996. Twenty-four cases (12%) were known to have died. One hundred and one cases were associated with travel, either abroad or in the United Kingdom. Two cases acquired infection in hospital, the smallest number ever reported, and the remaining 98 were presumed to have acquired infection in the community. Fifty-five (27%) of the 201 cases were linked to outbreaks or clusters and the remaining 146 (73%) were reported as single cases. Six outbreaks were associated with industrial premises. Twenty-two of the travel associated cases were part of three travel outbreaks and six clusters. The proportion of cases diagnosed by detection of urinary antigen has continued to increase and in 1996 this method of diagnosis was used for 43% of the cases. PMID- 9350093 TI - Serogroups/types and antibiotic resistance of referred isolates of Streptococcus pneumoniae: 1993 to 1995. AB - Surveillance of prevalent serogroups/types of Streptococcus pneumoniae and their susceptibility to antimicrobial agents is important for understanding the epidemiology of pneumococcal infections and for guiding empirical treatment. Current vaccines for prevention of pneumococcal infection utilise serotype specific antigens, so knowledge of the prevalence of particular serotypes is relevant to vaccine use and development. Five thousand seven hundred and ninety six isolates of S. pneumoniae from separate patients were serogrouped or serotyped by the Streptococcus and Diphtheria Reference Unit between 1993 and 1995. Antibiotic susceptibility testing was carried out by the Antibiotic Reference Unit on 3821 (65.9%) of these isolates. A total of 40 distinct serogroups/types, together with a small number of non-typable isolates, were noted over the three year period. The same five serogroups/types (6, 9, 14, 19, and 23) occurred most commonly in each year of the study, not only in the total population of isolates studied, but also in isolates obtained from blood or cerebrospinal fluid, and among isolates with antibiotic resistance. Ninety-six per cent of the isolates belonged to serogroups/types included in the currently available 23-valent capsular polysaccharide pneumococcal vaccine; the conjugate petna-, hepta-, and nonavalent vaccines covered 51%, 75%, and 80% of isolates respectively. The nonavalent vaccine offers the most promise as 74% of all blood and cerebrospinal fluid isolates and 90% of antibiotic resistant isolates belonged to serogroups or types included in this formulation. PMID- 9350094 TI - Audit of the implementation of selective neonatal BCG immunisation in south east London. AB - The implementation of a selective neonatal BCG immunisation policy adopted in south east London boroughs of Lambeth, Southwark, and Lewisham in 1992 has been audited. A survey conducted 18 months after the policy was implemented showed that only 11% of infants identified as eligible for neonatal BCG immunisation had been immunised. The results of the survey were fed back to neonatal units, which were encouraged to improve access to BCG immunisations for eligible infants. A second survey 17 months later showed that 14% of eligible infants had been immunised. Difficulties in applying complex selection criteria, rapid turnover of trained staff in acute units, and short neonatal stay were thought to contribute to the poor uptake of the selective programme delivered in the neonatal units. A community based BCG immunisation service has been commissioned to improve uptake. PMID- 9350096 TI - Increased incidence of hepatitis A in south east England. PMID- 9350095 TI - Lone parent families are an independent risk factor for lower rates of childhood immunisation in London. AB - The aim of this study was to determine associations between indicators of social deprivation and the uptake of primary immunisation in London. Correlation coefficients were calculated between immunisation coverage in London for each of the 28 inner and outer London district health authorities in November 1991 and a range of possible explanatory variables from small area statistics data from the November 1991 census. The proportions of children under 5 years of age, lone parent families, unemployed members of the workforce, domestic overcrowding, ethnic minorities, and unskilled workforce were correlated significantly with the coverage of primary immunisation for third dose diphtheria (D3) and pertussis (P3) at 12 months. A significant correlation with measles, mumps, and rubella (MMR) at 24 months existed only for lone parent families. Multiple linear regression weighted by population size was used to identify independent predictors of variation in immunisation cover. The proportion of lone parent families in each district health authority was the only significant independent risk factor consistently associated with variation in immunisation coverage for D3, P3, and MMR. The proportion of lone parent families explained 42% of the variation in coverage for D3 in November 1991. This study has identified lone parenthood as an important independent risk factor in London for failure to complete immunisation. PMID- 9350097 TI - Pathophysiological effects of laparoscopy: current knowledge. PMID- 9350098 TI - A 20-year experience with malpractice screening panels. PMID- 9350099 TI - Computer systems for hospitals and integrated health systems. PMID- 9350100 TI - Relationship between levels of soluble interleukin-2 receptors and the types and activity of vitiligo. AB - Levels of serum-soluble interleukin-2 receptors (serum sIL-2R) are thought to indicate the activation of immunocompetent cells, mainly lymphocytes. Elevated levels of serum sIL-2R have been observed in various infectious and autoimmune diseases and also after organ transplantation. It has been hypothesized that autoimmune mechanisms are involved in the pathogenesis of vitiligo. Therefore, we studied serum sIL-2R levels in relation to disease types and activity of vitiligo. We used sera separated from venous blood samples from 12 patients with dermatomally distributed type B vitiligo, 17 patients with non-dermatomally distributed type A vitiligo during the active stage, 9 patients with type A vitiligo during the inactive stage, and 12 normal control subjects. Serum sIL-2R levels were similar in type B vitiligo patients and the controls but were significantly elevated in patients with active type A vitiligo compared with controls and inactive type A vitiligo patients. It is suggested that the immune system is activated in patients with type A vitiligo during the active stage and that autoimmune mechanisms may play a role only in type A vitiligo. PMID- 9350101 TI - Human leukocyte interferon-alpha in a hydrophilic cream versus in a gel for the treatment of genital herpes in males: a placebo-controlled, double-blind, comparative study. AB - The aim of this double-blind, placebo-controlled, comparative study was to differentiate the clinical efficacy and tolerability of human leukocyte interferon-alpha incorporated (2 x 10(6) IU/g) in a hydrophilic cream and in a gel to heal males afflicted with first episodes of genital herpes. Patients (n = 60), aged 18-40 years (mean 23.2) with culture-confirmed diagnosis of herpes genitalis were randomized to three parallel groups. Each patient was allocated a precoded 40-g tube, containing either preparation or placebo. Cream or gel was applied three times daily for 5 consecutive days. The duration of the active treatment was two weeks. Patients were examined after 48 hours in initial treatment, and thereafter two times a week. A reepithelialized lesion with some residual erythema was recorded as healed. The study demonstrated that patients treated with leukocyte interferon-alpha cream had both significantly shorter mean duration of lesions than gel and placebo recipients (5.3 days vs. 8 days, 13 days respectively; p < 0.001) and a higher number of healed patients (80% vs. 55%, 20% respectively; p < 0.001). Of the 60 patients, 49 (82%) complained of no drug related side effects. Eleven patients predominantly in the cream/gel groups reported non-objective transitory increase in their body temperature (> 38 degrees C) with moderate headache, malaise and myalgia. The study was followed-up for 24 months after the first day of the treatment, and out of 31/60 cured patients, 4 had a relapse after 18 months. In conclusion the study affirmed that human leukocyte interferon-alpha (2 x 10(6) IU/g) in a hydrophilic cream is more efficacious than its incorporation in gel or placebo, thus suggesting that leukocyte interferon-alpha in a hydrophilic cream, with a profile of non objective mild to moderate drug-induced indications, may be considered an alternative and effective treatment modality to cure male patients afflicted with first episodes of genital herpes. PMID- 9350102 TI - Lupus panniculitis treated by a combination therapy of hydroxychloroquine and quinacrine. AB - Lupus erythematosus panniculitis (LEP) is an unusual clinical variant of lupus erythematosus (LE) in which the cutaneous inflammatory reaction occurs primarily in the deeper corium. The common clinical features of LEP includes asymptomatic, firm, sharply defined nodules. The histologic findings are characterized by nonspecific panniculitis composed of lymphoid cells, plasma cells, and histiocytes with varying degrees of necrobiotic changes with fibrinoid deposits. In our case, a 24-year-old male patient visited our clinic with non-tender, hard, plaque-like lesions and overlying erythema on the left zygomatic, nasal, and submandibular area. Histopathologic and direct immunofluorescent findings of the lesion were compatible with LEP. His skin lesions waxed and waned with systemic steroid or hydroxychloroquine therapy. He has responded well to a combination therapy of hydroxychloroquine and quinacrine. PMID- 9350103 TI - A verrucous lesion on skin grafted after necrotizing fasciitis in a diabetic patient successfully treated with combined topical 5-FU and tacalcitol. AB - Many complications of diabetes mellitus involve the feet. These include infections, neuropathy, vasculopathy, and poor wound healing. Neuropathy causes chronic pressure or friction on an area of sensory loss and occasionally causes verrucous skin lesions. We describe a diabetic patient, complicated by necrotizing fasciitis, who developed a verrucous skin lesion on a skin graft site. The verrucous skin lesion was treated successfully with combined topical 5 fluorouracil and vitamin D3 application. PMID- 9350104 TI - Neonatal lupus erythematosus occurring in identical twins. AB - The patients were one-month-old identical twins. Scaly erythema was noted mainly on the trunk, face, and scalp of one twin starting about three weeks after birth and starting about two weeks after birth in the other. The patients' courses were observed without treatment; the eruptions tended to disappear two months after birth. The mother had a past history of transient facial erythema. Both twins and mother were positive for antinuclear antibody, anti-SS-A antibody, and anti-SS-B antibody. The histopathological findings corresponded to those of discoid lupus erythematosus (DLE). The class of HLA typing revealed Cw3 in both twins, which is frequently observed in neonatal lupus erythematosus (NLE), and A24 in the mother, which is frequently observed in mothers of babies with NLE. PMID- 9350106 TI - A case of pseudovascular adenoid squamous cell carcinoma of the skin with spindle cell pattern. AB - We report a case of pseudovascular adenoid squamous cell carcinoma which was in the form of a dome-shaped, cherry red tumor on the right cheek of an 86-year-old Japanese woman. Histologically, two types of atypical cells were identified; round cells in the upper part and spindle cells in the lower part. In the upper part, inter-anastomosing sinusoid-like pseudolumina were observed between the cords of the tumor cells. Immunohistochemically, the tumor cells did not express Factor VIII-related and CD34 antigens or bind Ulex europaes I agglutinin, except for only one anti-cytokeratin antibody. Ultrastructurally, the tumor cells contained tonofilaments and desmosomes and represented acantholysis. From the electron microscopy, possible role of capillary hyperpermeability on the acantholytic process of the neoplastic cells was suggested. PMID- 9350105 TI - Koebner phenomenon in an ANCA-positive patient with pyoderma gangrenosum. AB - A male with pyoderma gangrenosum is reported. The clinical and histological features were typical. The initial lesions resolved with characteristic cribriform scars. A few days after the complete recovery, he developed several necrotizing focal lesions localized to the scarred areas. A further histological examination revealed a granulation tissue rich in neutrophils and signs of necrotizing vasculitis. We found a high titer of circulating perinuclear antineutrophil antibodies (p-ANCA), which are a serological marker for various systemic diseases. An immunological circulating factor has been repeatedly suggested to be the "primum movens" of pyoderma gangrenosum. We discuss the unusual clinical presentation interpreted as a Koebner phenomenon and the possible role of immune factors in enhancing circulating-endothelial cell interactions in relation to the pathogenesis of pyoderma gangrenosum. PMID- 9350107 TI - Zeis gland carcinoma. AB - Sebaceous carcinoma of the Zeis gland of the cilia is a rare malignant tumor. A 90-year-old Japanese man with a Zeis gland carcinoma on the left upper eyelid is reported. The tumor was a 12 x 3-mm, reddish, partially yellow, papillomatous nodule accompanied by loss of several lashes. Histologically, two types of tumor cells, a dark basaloid cell and a larger pale cell, showed lobular and comedo acinar growth patterns. The carcinoma did not recur during the 8 months following surgery; however, the patient died of gastric carcinoma. PMID- 9350109 TI - Rudimentary meningocele of the scalp. AB - A rudimentary meningocele, a variant of primary cutaneous meningioma, was seen on the scalp of a 9-month-old Japanese boy. Clinically, the lesion on the left parietal area was round, about 1.6 cm in diameter, alopecic, and slightly elevated. Histologically, the lesion, located from the dermis to the subcutis, consisted of scattered foci of meningothelial cells, an anastomosing network of empty spaces with psammoma bodies and collagen bodies, and small vessels. Immunohistochemically, the meningothelial cells were positive for vimentin and desmin. Ultrastructurally, they had elongated cytoplasmic processes, intermediate filaments in the cytoplasm, and desmosomal junctions. PMID- 9350108 TI - Clear cell acanthoma developing in epidermal nevus. AB - A 33-year-old Japanese woman presented with a black papule on a pigmented lesion which had been on her right thigh since her early childhood. A hematoxylin-eosin stained section revealed a sharply demarcated, acanthotic epidermis composed of enlarged clear cells, which stained positively for epithelial membrane antigen and negatively for carcinoembryonic antigen. With antikeratin antibodies, the tumor cells stained for AE1 and AE3, but did not stain for CAM5.2. They contained abundant glycogen. Histologically, we diagnosed the case as a clear cell acanthoma which developed in the pre-existing epidermal nevus. This is the second such case in the literature. PMID- 9350110 TI - Erythema nodosum leprosum in histoid leprosy: a case report. AB - A 28-year-old married female developed histoid papules and nodules de novo over her face, extremities, back, buttocks and thighs. She had developed erythema nodosum leprosum lesions without any antileprosy treatment. Histopathology from a histoid nodule showed well defined nodular collections of plump, spindle-shaped histiocytes. A few globi were also seen with Ziehl Neelson staining. Leucocytoclastic vasculitis was present in the ENL lesion. The CD4:CD8 ratio was 1.5:1. PMID- 9350112 TI - Sexually transmitted diseases in Rhode Island: a panoply of familiar as well as previously unrecognized challenges. PMID- 9350111 TI - Is there any treatment of leukotrichia in stable vitiligo? PMID- 9350113 TI - The Salvation Army and phossy jaw. PMID- 9350114 TI - Sexually transmitted diseases: the hidden epidemic. PMID- 9350115 TI - Sexually transmitted diseases in Rhode Island: past, present, and future. PMID- 9350116 TI - Pelvic inflammatory disease in Rhode Island: epidemiology, diagnosis and therapy. PMID- 9350117 TI - Pathogenesis of human papillomavirus infection of the genitals. PMID- 9350118 TI - HIV infection in Rhode Island adolescents. PMID- 9350119 TI - Human immunodeficiency virus infection and sexually transmitted diseases. PMID- 9350120 TI - Three decades of research on sexual behavior and sexually transmitted pathogens in college students. PMID- 9350121 TI - Changes in sexual behavior related to HIV/AIDS knowledge. PMID- 9350122 TI - Proposed prostate cancer screening recommendations. PMID- 9350123 TI - Animated suspension in the intercostal state: a light-hearted ribbing. PMID- 9350124 TI - Point of view: from Gregor Mendel to coronary atherosclerosis. PMID- 9350126 TI - A promising partnership. PMID- 9350125 TI - The Morocco cooking scandal of 1959. PMID- 9350127 TI - When symptoms defy diagnosis. PMID- 9350128 TI - Treating the batterer. Help for men who hurt. PMID- 9350129 TI - Diagnostic and treatment guide for domestic violence. PMID- 9350130 TI - Kids at risk. PMID- 9350131 TI - Domestic violence. Reality for some of your toughest patients. AB - Until recently, domestic violence was considered a criminal justice or social service problem. However, physicians see victims in emergency settings and clinics with complaints and symptoms that go beyond physical injuries. A study by a Minnesota health plan shows that, on average, a victim of domestic violence costs the health care system $1,434 more per year than a nonvictim. This article discusses the prevalence of domestic violence and the variety of presentations. Guidelines for screening, what to do with a positive response, when to call the police, and how to document and code are reviewed, as are issues unique to older victims of domestic violence. PMID- 9350132 TI - The physician's role in tracking injuries: the importance of E-codes. PMID- 9350133 TI - Screening for domestic violence in a rural family practice. AB - This study evaluates the effectiveness of a screening form used to assess the incidence of current and past domestic abuse among patients seen in a rural family practice. Two of nine family physicians screened most adult women they saw at a rural primary care clinic for three months in early 1996. Of the 280 women who completed surveys, 94(34%) reported experiencing physical or emotional abuse at some time in the past. Twenty-three (8%) reported physical abuse within the past year. Fourteen women (5%) reported being currently afraid of their partner or someone else. Although current or past abuse was seen across all groups, women reporting abuse were more likely to be unmarried, younger, and on medical assistance. They were also more likely to have children at home. We concluded that the prevalence of women reporting current abuse in a rural family practice is sufficient to warrant mass screening. PMID- 9350134 TI - Legal issues and remedies for victims of domestic violence. PMID- 9350135 TI - The physical location of genes cdc2 and prh1 in maize (Zea mays L.). AB - A biotin-labelling in situ hybridization technique was first used to physically map two single copy genes, cdc2 and prh1, in maize. These two genes are metabolically interrelated genes. The full-length cDNA clones cdc2ZmA and ZmPPI of genes cdc2 and prh1 were adopted as the probes. They are 1.3 and 1.6 kb in size, respectively. Clone cdc2ZmA was physically mapped on the long arm of chromosomes 4, 8, and 9. The percent distances from centromere to detection site were 57.9 +/- 2.7, 28.4 +/- 1.5, and 88.2 +/- 3.3. The detection rate was 19.2%. Clone ZmPPI was physically mapped on the long arm of chromosomes 4, 6, and 8. The percent distances were 53.6 +/- 1.2, 60.8 +/- 2.9 and 17.1 +/- 1.6. The detection ratio was 18.5%. The technique of chromosome ISH and the relationship between the location and function of these two genes have been discussed. PMID- 9350136 TI - A specific alpha-tubulin is associated with the initiation of parthenogenesis in 'Salmon' wheat lines. AB - The 'Salmon system' consists of isogenic but alloplasmic wheat lines with either sexual or autonomous embryo development. Using two-dimensional gel electrophoresis these lines have been screened for proteins potentially involved in the initiation of parthenogenesis. A temporally altered expression of the polypeptide 'P 115.1' in the sexual and parthenogenetic 'Salmon' lines seems to be related with the autonomous embryo formation. Around anthesis when most of the egg cells begin the parthenogenetic development, the polypeptide 'P 115.1' was present in ovaries of the parthenogenetic lines but not in ovaries of the sexual line. Moreover, this polypeptide is only expressed in the ovaries of amphidiploid parthenogenetic plants containing differentiated embryo sacs. It is absent from ovaries of the analogous polyhaploid plants, which lack any embryo sac structure within their ovules. Furthermore, the polypeptide was neither detectable in meristematic tissue of root tips nor in leaves. N-terminal amino acid sequencing identified 'P 115.1' as an alpha-tubulin. Thus, 'P 115.1' apparently represents an embryo sac-specific isoform of alpha-tubulin involved in the initiation of embryo development. PMID- 9350137 TI - Correlations among size-related traits affected by chromosome inversions in Drosophila buzzatii: the comparison within and across environments. AB - Genetic variation in correlations among size-related traits of head, thorax, and wings was examined in Drosophila buzzatii, by comparing the pattern of the Phenotypic Correlation Matrix (Rp) between inversion karyotypes of the second chromosome. No similarity in Rp was observed between some karyotypes in a natural population. The pattern of Rp in wild-reared heterokaryotypes, but not in homokaryotypes, was similar to the whole population represented by laboratory reared flies. While phenotypic correlations in wild-reared flies were found to be larger than in laboratory-reared flies, similarity in the pattern of Rp was very high for one homokaryotype reared in both environments: the relatively homogeneous lab environment and the more variable field environment. While no such a similarity across environments was detected between different karyotypes, the pattern of Rp was similar for a same homokaryotype in different populations. Thus, the lack of karyotypic similarity in Rp is, at least partially, genetic. These results indicate that chromosomal inversions are factors affecting genetic correlations among traits known to be phenotypically correlated with adult fitness components in this species. PMID- 9350138 TI - The effect of polymorphic inversions on body size in two natural populations of Drosophila buzzatii from Argentina. AB - Previous works in a colonized and an original population of Drosophila buzzatii have shown a consistent relationship between the inversion polymorphism and thorax length, a measure of body size. However, the populations studied in those reports share a close genealogical relationship as suggested by several lines of evidence. In the present paper, we revisit this issue by analysing the correlation between second chromosome arrangements and thorax length in two Argentinian natural populations (Termas de Rio Hondo and Arroyo Escobar) from different biogeographic areas with different host plants. Our findings are: (1) inversion frequencies were significantly different between populations; (2) the mean thorax length of flies collected in both populations was not significantly different; and (3) we obtain confirming evidence that flies carrying 2st, the ancestral gene order, have on average a smaller body size than those carrying the derived arrangements (2j and 2jz3). These results suggest that the biometrical effect of inversions on body size previously described are due to genetic differences between arrangements and not to the close historical relationship between the populations studied in previous reports. PMID- 9350139 TI - Polymorphism of alcohol dehydrogenase genes in alcoholic and nonalcoholic individuals from Valencia (Spain). AB - Polymorphisms in the two variable ADH loci (ADH2 and ADH3) were analyzed in two groups (alcoholics and nonalcoholics) from a Spanish population. The frequencies were similar to those reported for other Causcasian groups. ADH2-1 and ADH3-1 genotypes were more frequent in alcoholics than in nonalcoholics, but the differences were not significant. PMID- 9350140 TI - Cytogenetic evaluation of 20 primary breast carcinomas. AB - Chromosome analysis was performed on samples from 20 Brazilian patients with breast cancer. All the samples were from untreated patients who presented the clinical symptoms for months or years before surgical intervention. Six cases showed axillary lymph node metastases. Clonal chromosome abnormalities were detected in all cases. The numerical alterations most frequently observed involved the loss of chromosomes X, 19, 20, and 22 followed by gain of chromosomes 9 and 8. Among the structural anomalies observed, there was preferential involvement of chromosomes 11, 6, 1, 7, 3, and 12, supporting previous reports that these chromosomes may harbour genes of importance in the development of breast tumors. Two cases with a family history of breast cancer had in common total or partial trisomy 1. PMID- 9350141 TI - Variation in coxII intron in the wild ancestral species of wheat. AB - To study the maternal lineage and evolution of polyploid species of wheat, variation in mitochondrial DNA was investigated in Triticum and Aegilops by PCR aided RFLP analysis. A 1.3 kb region containing the intron of coxII was studied using 20 accessions from five species of Sitopsis section of Aegilops, one species of Einkorn wheat, four species of tetraploid wheat, and one species of common wheat. Only three restriction site changes and a single deletion/insertion were found among 884 restriction fragments surveyed. This fact suggests the highly conserved nature of this region within Triticum and Aegilops. Four haplo types were recognized in coxII intron. A parsimonious relationship indicated that three haplo-types were independently derived from one prototype which was found in wild Einkorn and Aegilops species except for Ae. speltoides. All but one accession of Ae. speltoides possessed a derivative haplo-type, common in Timopheevi wheat. The result supported the hypothesis that Ae. speltoides donated the G genome to Timopheevi wheat; however did not agree with that Ae. speltoides was the B genome donor to the Emmer and common wheat. PMID- 9350142 TI - Physical localization of rRNA genes by two-colour fluorescent in-situ hybridization and sequence analysis of the 5S rRNA gene in Phalaris coerulescens. AB - The 18S-5.8S-26S rDNA and 5S rDNA loci have been mapped physically by fluorescent in-situ hybridization to the chromosomes of Phalaris coerulescens. The biotin labelled heterologous 18S-5.8S-26S rRNA probe (pTa71) detected one locus, which corresponded to the secondary constriction (nucleolar organizer) on the long arm of the satellited chromosome II. The homologous 5S rDNA probe (Bam2.12) detected two pairs of 5S rRNA gene clusters which were localized at two different non satellited chromosomes, one near the telomere on the short arm of the chromosome I, which is the largest chromosome of the complement, and the other about 42% out on the long arm of the chromosome III. A BamHI fragment containing the 5S rRNA gene, has been isolated and characterized. The 5S rDNA repeat unit is 309 bp in length, consisting of 121 bp highly conserved coding region and 188 bp variable spacer region. The karyotype of Phalaris coerulescens is characterized by the similar size of chromosomes within the group 2, group 3, or group 4. This study represents the first step towards the understanding the genome organization of Phalaris coerulescens and provides reliable markers for chromosome identification in this grass, an important species as a model system for the study of self incompatibility in grasses. PMID- 9350143 TI - Cardiac contractility in noninsulin dependent diabetes mellitus evaluated using the relation between endsystolic wall stress and velocity of circumferential fiber shortening. AB - The relation between left ventricular (LV) endsystolic wall stress (sigma es) and rate-corrected velocity of circumferential fiber shortening (Vcfc), which is independent of heart rate (HR) and loading conditions has previously been used to assess cardiac contractility in insulin dependent diabetes mellitus (IDDM). This study is the first report in which this relation has been utilized with data obtained by echocardiography in addition to the traditional indices, to evaluate the cardiac function in asymptomatic, middle-aged patients with noninsulin dependent diabetes mellitus (NIDDM) at baseline and during dobutamine stimulation. There were 16 NIDDM patients in our study and these patients were classified into 2 groups. Group 1 consisted of 11 patients without microvascular complications. Group 2 consisted of the remaining 5 patients with microvascular complications. Ten age- and sex-matched normal subjects were enrolled as a control group. At baseline, diabetic patients tended to have a faster HR and a greater LV enddiastolic dimension, though these values were not significantly different from the normal subjects. Ejection fraction (EF) in group 1 was significantly higher than that of the normal controls (73 +/- 2% vs 65 +/- 2%, p < 0.005). Mitral inflow pattern was normal (E/A > 1) in the normal subjects (1.11 +/- 0.06), but reversed in group 1 (0.87 +/- 0.07) and group 2 (0.95 +/- 0.12). Isovolumic relaxation time corrected for HR (IVRTc) and the slope of relation between sigma es and Vcfc were similar among the 3 groups. Comparing Vcfc at 50 g/cm2 of sigma es, it tended to increase from the normal subjects (0.883 +/- 0.057 cir/sec) to 0.969 +/- 0.048 in group 1 and 1.034 +/- 0.101 in group 2, though this result was not statistically significant. During dobutamine stimulation, EF increased and IVRTc shortened significantly only in the normal subjects. E/A became normalized in both diabetic groups. The increment in Vcfc representing cardiac reserve of contractility was significantly lower in the diabetics (0.110 +/- 0.040 in group 1 and 0.057 +/- 0.043 in group 2) than in normal subjects (0.244 +/- 0.044). In conclusion, using the index of relation between sigma es and Vcfc, the cardiac contractility of NIDDM was not impaired at baseline, and even had a tendency to increase. However, during dobutamine stimulation, the inadequate reserve of contractility was exposed, especially in those patients who had microvascular complications. These results indicate the importance of controlling diabetes, not only to prevent the development of microvascular complications but also to preserve cardiac function. PMID- 9350144 TI - Additional data on body surface potential maps of ventricular repolarization in normal adults. AB - To date most published studies on normal basic data of the potential distribution of cardiac activity have been restricted to ventricular depolarization. Only a few papers have dealt with ventricular repolarization in normal subjects. This is an attempt to establish the basic data of the body surface potential maps (BSPM) of ventricular repolarization in normal adults. BSPM of ventricular repolarization utilizing 87 electrodes through the heart potential map system designed by Toyama et al. were studied in 50 normal Chinese male adults. The following information on BSPMs was obtained: (1) In the initial phase of ventricular repolarization, the potential maximum was located on the precordial area and the potential minimum appeared on the right-superior portion of the back; (2) The movement of potentials was counterclockwise and stable during the later portion of the T loop; (3) The magnitudes of the potentials changed in a spindle pattern with the drifting of ventricular repolarization. They first increased and then decreased with the largest value being around the peak period of the T wave; (4) The absolute value was greater in the potential maximum than the potential minimum throughout ventricular repolarization. There was no "reversal" potential distribution pattern. This parameter may be of importance clinically; (5) There were usually multiple potential maxima and multiple potential minima during the ST segment and early phase of the T wave; (6) The largest potential maximum and potential minimum were 0.93 +/- 0.28 mV and 0.35 +/ 0.19 mV, respectively. The potential maximum and potential minimum at the peak T wave were 0.91 +/- 0.23 mV and 0.35 +/- 0.17 mV, respectively. Obviously, this study offers valuable basic data on the BSPM in normal adults and will be helpful to our understanding of the BSPM in various heart diseases. PMID- 9350145 TI - Transesophageal echocardiographic prediction of initially successful electrical cardioversion of isolated atrial fibrillation. Effects of left atrial appendage function. AB - Left atrial appendage (LAA) flow velocities prior to electrical cardioversion were recorded using transesophageal pulsed Doppler echocardiography to predict initially successful cardioversion of isolated atrial fibrillation (AF). Patients with AF were placed into either a success group (19 patients) in which sinus rhythm was maintained for at least 2 days or a failure group (12 patients). The duration of AF was shorter in the success group. The maximum left atrial diameter was the same for the two groups. The maximum LAA area was smaller in the success group. The maximum forward and backward LAA velocities were greater in the success group, as were the mean forward and backward LAA velocities. In the patients with mean LAA flow velocities greater than 19 cm/sec, the success of cardioversion could be predicted with high sensitivity (80%) and specificity (88%). We conclude that the duration of AF, the maximum LAA area, and LAA flow velocities prior to cardioversion predict the initial recovery of sinus rhythm for isolated AF. PMID- 9350146 TI - The effect of cigarette smoking during pregnancy on cord blood lipid, lipoprotein and apolipoprotein levels. AB - We examined the relationship between maternal smoking during pregnancy and serum lipid, lipoprotein and apolipoprotein levels in newborns. Serum concentrations of total cholesterol, triglyceride, high density lipoprotein cholesterol, low density lipoprotein cholesterol, apolipoprotein A-1, apolipoprotein B and lipoprotein (a) were assessed in blood samples from 38 mothers who were smokers and their newborns obtained at delivery and compared to blood sample from 42 nonsmokers and their newborns. As compared with newborns of nonsmoker mothers, newborns of smoker mothers showed a lower mean level of high density lipoprotein cholesterol (21 versus 26 mg/dl, p < 0.01), a higher total cholesterol to high density lipoprotein cholesterol (4.7 versus 3.7, p < 0.01), a higher low density lipoprotein cholesterol to high density lipoprotein cholesterol ratios (3.2 versus 2.3, p < 0.05), a lower mean level of apolipoprotein A-1 (105 versus 129 mg/dl, p < 0.01) and a higher apolipoprotein B to apolipoprotein A-1 ratio (0.44 versus 0.3, p < 0.01). These parameters were also different between smoker and nonsmoker mothers. There were no significant differences in TC, TG, LDL-C, Apo B and Lp (a) values between the two newborn groups. These data suggest that maternal smoking during pregnancy markedly affects lipid metabolism in the fetus. PMID- 9350147 TI - Reversal of early metabolic dysfunction in hypertensive rat left-ventricular myocytes by angiotensin-converting enzyme inhibition. AB - We evaluated the effects of angiotensin-converting enzyme (ACE) inhibition on metabolic changes in myocardial organelles, myocardial hypertrophy, and interstitial fibrosis in the early stage of hypertension. An ACE inhibitor, imidapril (2.5 mg/kg per day), a calcium-channel blocker, diltiazem (30 mg/kg per day), or vehicle was given to spontaneously hypertensive rats (SHRs) from 10 to 18 weeks of age. Single myocytes were isolated enzymatically from the left ventricles of these SHRs and normotensive Wistar-Kyoto (WKY) controls at 18 weeks of age. In single ventricular myocytes, enzyme activities in the sarcoplasmic reticulum (SR) and the sarcolemma (SL) and the mitochondrial respiratory control ratio (RCR) were determined. In 18-week-old SHRs receiving vehicle, myocardial hypertrophy and interstitial fibrosis developed, and SR Ca2+ AT-Pase activity and the mitochondrial RCR were significantly lower and SL Na+, K(+)-ATPase activity was significantly higher than in age-matched WKYs. However, compared with diltiazem, imidapril was better able to prevent the development of myocardial hypertrophy and interstitial fibrosis, to improve SR Ca(2+)-ATPase activity and the mitochondrial RCR, and to increase SL Na+, K(+)-ATPase activity. These results suggest that ACE inhibition can prevent the development of morphologic changes associated with hypertension-induced left ventricular remodeling, such as myocardial hypertrophy and interstitial fibrosis, and can counteract ongoing dysfunction of organelle metabolism early in the development of hypertension. PMID- 9350149 TI - Thrombectomy for left ventricular protruding thrombi in a patient with dilated cardiomyopathy. AB - Left ventricular thrombus is one source of cardiogenic embolism. The protruding, mobile type is the highest risk subgroup but is rarely encountered. Thrombectomy is one choice of therapy, and variously recommended based primarily on the experience with myocardial infarction. We report a rare case of successful left ventricular thrombectomy for two protruding, mobile thrombi in a patient with idiopathic dilated cardiomyopathy in order to prevent repeat embolization. PMID- 9350148 TI - The effects of dopamine, dobutamine and amrinone on mitochondrial function in cardiogenic shock. AB - The impairment of mitochondrial in non-infarcted myocardium under cardiogenic shock complicated by acute myocardial infarction was studied. We induced acute myocardial infarction in dogs by ligating the circumflex branch of the left coronary artery (LCX). On basis of left ventricular systolic pressure (LVPs) after 60 minutes, we divided the dogs into two groups: a group in which LVPs fell to below 70% of the pre-LCX ligation level, and a Control group in which LVPs remained more than 90%. The former group was further divided into four subgroups, depending on infusion of dopamine, dobutamine, amrinone or saline after 90 minutes. Mitochondria were prepared and mitochondrial respiratory activity determined. In the Saline group, hemodynamics became reduced to less than 70% of the preligation level after 120 minutes, however, in the Dopamine and Dobutamine groups, hemodynamics became restored to the preligation level. In the Amrinone group, LVPs decreased slightly, while cardiac output, LV Max. dp/dt and myocardial blood flow increased. In the Saline group, mitochondria in the non infarcted myocardium functioned at a lower level of activity than that of the Control group. However, in the Dopamine, Dobutamine, and Amrinone groups, the mitochondria functioned at a higher level. Electron microscopy revealed mitochondrial damage in the Saline group only. The results indicate that an energy production disorder in the non-infarcted myocardium may have pathogenetic implications in cardiogenic shock associated with acute myocardial infarction, while dopamine, dobutamine, and amrinone improve mitochondrial function, and ultimately improve cardiac function. PMID- 9350150 TI - Exercise-induced neurally mediated syncope. AB - We describe a 16-year-old female referred for evaluation of syncope associated with competitive long distance running. She had experienced 4 episodes of syncope during competitive long distance racing. The syncope associated with marked bradycardia and asystole was demonstrated by head-up tilt testing without isoproterenol infusion. Oral propranolol therapy failed to prevent the syncope. Oral disopyramide therapy, however, prevented the syncope induced by both head-up tilt testing and competitive long distance racing. Caution should be urged in evaluating athletes with syncope, especially in the pediatric age group, because the cause of the syncope may result from life-threatening disorders such as cardiomyopathy, long QT syndrome, or exercise-induced arrhythmias. The head-up tilt test is an important diagnostic tool for the evaluation of exercise associated syncope. PMID- 9350151 TI - Rhabdomyolysis, acute renal failure and hepatopathy induced by lovastatin monotherapy. AB - Lovastatin is a popular drug for the treatment of hypercholesterolemia. Few serious complications are associated with its use although rhabdomyolysis with renal failure has been reported when lovastatin is combined with cyclosporine or other drugs following transplantation. We report the case of a patient who developed hepatopathy and rhabdomyolysis accompanied by acute renal failure after 4 weeks of lovastatin monotherapy. The pathogenesis and treatment of these complications are also discussed. PMID- 9350152 TI - The use of anticholinergics in asthma. AB - Anticholinergic medications have been accepted as an important treatment modality in chronic bronchitis and chronic asthma, but their use in acute asthma is more controversial. A brief historical context of anticholinergics is given. The innervations of the lung that govern bronchoconstriction and bronchodilatation are reviewed. The pharmacological and neurological properties of anticholinergics make them excellent modalities for treatment of asthma. The benefits of anticholinergics in acute asthma, exercise-induced asthma, nocturnal asthma, and psychogenic asthma are reviewed. The use of anticholinergics in anaphylaxis with beta-blockade is examined. PMID- 9350153 TI - Occupational allergic disease in cereal workers by stored grain pests. AB - It is well known that workers occupationally exposed to grain dust have a high prevalence of respiratory symptoms, but their pathogenesis remains obscure when sensitization to cereal flour cannot be demonstrated. Storage mites, tenebroids, and cockroaches are stored-grain pests found in grain and cereal products frequently in our area, where the cereal industry is the most important industry. An epidemiological analysis of sensitization of these stored-grain pests was performed on 4379 patients residing in an area of cereal industries. Fifty grain workers were selected for in vivo diagnostic tests with nine genera of mites, Tenebrio molitor and Blatta orientalis. Specific IgE antibodies to the extracts were demonstrated by prick tests and RAST. Association between respiratory symptoms and occupational exposure was confirmed by challenge tests (specific and methacholine). The prevalence of mite sensitization in the total sample studied (4379) was 18.96% (SEM 0.58, 95% CI 16.93-19.19). The prevalence of sensitization to storage mites among mite-sensitive patients was 11.88% (SEM 1.15, 95% CI 9.63 14.3). Among the 50 selected patients the most frequent sensitization was that to Dermatophagoides pteronyssinus (58%), followed by Dermatophagoides frinae (48%), Lepidoglyphus destructor and Tyrophagus putrescentiae (38%), Blomia kulagini (34%), and Acarus siro and Chortoglyphus arcuatus (24%). In addition, 22% of the patients presented negative prick tests and RAST for Dermatophagoides species with positive test to storage mites. Fifty percent of the 50 patients were sensitizated to Tenebrio molitor (SEM 0.7, CI 95% 36-64), and 36% to Blatta orientalis (SEM 0.67, CI 95% 23-49). The identification of mites, tenebroids, and cockroaches in dust samples yields useful data for the diagnosis of our patients. PMID- 9350154 TI - Asthma training in third-year medical students. AB - To assess the educational experiences of physicians-in-training with asthma patients, we had medical students complete asthma surveys at the beginning and end of their internal medicine clerkship (IMC). At the beginning of the IMC, all students received a 1-hr asthma lecture and half of the students received a compilation of pocket cards containing many of the algorithms from the National Heart, Lung, and Blood Institute asthma guidelines. We found that students had relatively few encounters with asthmatic patients during the IMC. Students were good judges of asthma severity but performed poorly on survey questions pertaining to asthma treatment. Confidence in treating and assessing patients improved by the end of the IMC, but remained low. We conclude that the usual 1-hr lecture and limited contact with asthma patients during the IMC may be in adequate to train students to care for patients with asthma. PMID- 9350155 TI - Patient referrals to a multispecialty asthma clinic. Asthma Care Center Clinical Consortium. AB - A total of 656 patients with asthma have been referred to a multispecialty pediatric asthma clinic for evaluation; 52 (7.9%): mild; 406 (61.9%): moderate; 177 (27%): severe; and 21 (3.2%): incomplete data. No significant differences in demographics or payer source were observed across disease severity levels. Only 25% of the patients with primary care providers were referred by these practitioners. Over 20% of the mild asthmatics were using inhaled bronchodilators regularly. Only 40% and 50% of the moderate and severe asthmatics, respectively, were using inhaled bronchodilators regularly, and only 19% and 36%, respectively, were on maintenance inhaled corticosteroids. Pressures to reduce subspecialty services may place some of these asthma patients at increased risk for complications from this chronic lung disease. PMID- 9350156 TI - Evaluation of an educational program for asthma control in adults. AB - We have developed a 6-month educational plan associated with outpatient follow-up and special clinical care for asthmatic patients in a deprived population, with serious socioeconomic problems and a very low level of education. The objective was to determine the effects of the program on clinical asthma outcomes, lung function, and quality of life. Forty patients were enrolled in the program with a regular schedule of outpatient visits, and 31 finished the 6-month intervention, which included information about asthma, instruction in appropriate use of medication, training in metered-dose inhaler technique, how to identify and control asthma triggers, how to use symptom diary cards, and how to recognize early signs of deterioration. Patients included 8 males and 23 females, 47.8 +/- 16.5 years old, with 77.4% elementary school education and 22.6% illiterate, and an average monthly income of around $450. After the 6-month program there was a significant change in asthma control with a reduction in asthma emergency visits and hospitalization, reduction of score symptoms, and improvement in quality of life. Based on the results, educational programs are recommended and should be adapted to the socioeconomic and cultural characteristics of the target population. PMID- 9350157 TI - Changes of soluble ICAM-1 levels in serum and bronchoalveolar lavage fluid from patients with atopic bronchial asthma after allergen challenge. AB - Leukocyte-endothelial cell interaction is essential for leukocyte infiltration into inflammatory sites. Initiation of adhesion is through the up-regulated expression of adhesion molecules in the endothelium or epithelium and the activation of adhesion molecules on leukocytes. To our knowledge, there have been few reports concerning soluble intercellular adhesion molecule-1 (sICAM-1) in patients with atopic bronchial asthma after allergen challenge. If the levels of sICAM-1 vary between bronchial asthma patients and normal controls, this variance would be useful to assess the state of this disease. Therefore, we measured the levels of sICAM-1 in sera from 17 patients with atopic bronchial asthma and normal control subjects. Levels of sICAM-1 in sera from bronchial asthma patients in prechallenge conditions were higher than in normal control subjects. Levels of sICAM-1 in sera from bronchial asthma patients 8 hr after challenge were higher than those in sera obtained during prechallenge periods. sICAM-1 levels in bronchoalveolar lavage (BAL) fluids from bronchial asthma patients 8 hr after challenge were higher than at 30 min after challenge. These results suggest that higher levels of sICAM-1 in sera and BAL fluids reflect the up-regulation of ICAM 1 expression in allergic bronchial asthma and these high levels may contribute to the pathogenesis of atopic bronchial asthma. PMID- 9350158 TI - Aspirin-induced asthma as a risk factor for asthma mortality. AB - We have recently reported severe airway obstruction and bronchial hyperresponsiveness (BHR) in fatal asthma compared with patients without a history of near-fatal asthma. Based on these findings, we evaluated whether aspirin-induced asthma (AIA) is a risk factor for asthma mortality by analyzing pulmonary function and BHR. FEV1.0% and methacholine inhalation threshold were significantly lower in the fatal asthma group compared with the AIA and non-AIA groups. However, there were no significant differences between the AIA and non AIA groups. Our results suggested that AIA is not a risk factor for asthma mortality if avoidance of aspirin and aspirin-like drugs is assured. PMID- 9350160 TI - Nasal Pathophysiology impacts bronchial reactivity in asthmatic patients with allergic rhinitis. PMID- 9350161 TI - Floating patterns of metered dose inhalers. AB - As long as metered dose inhalers have existed, patients have sought a reliable method to determine if a given canister was still potent. Concerning beta agonists, the answer to this question may be lifesaving. Issues of compliance have made dating canisters or counting doses impractical. Likewise, previous claims of floating characteristics are unreliable. In tap water, we float-tested 13 commonly used inhalers three times each, observing variations as they were incrementally actuated, emptying their contents. One essential pattern was observed. Almost all prescription-size canisters sink when full; all float by the time one-third of their contents is gone. Orientation of prescription-size canisters changes in a distinct pattern especially near 90% depletion. Sample size canisters showed some variance. Results suggest that the pharmaceutical industry should include individual floating characteristics as part of the package insert as they provide a reproducible means of gauging contents. PMID- 9350159 TI - Symptomatic improvement following emergency department management of asthma: a pilot study of intramuscular dexamethasone versus oral prednisone. AB - Systemic corticosteroid therapy is an established adjunct to beta-adrenergic medications in acute exacerbations of asthma. To date, no study has defined the role of long-acting intramuscular preparations of corticosteroids in pediatric patients with asthma. A pilot study was conducted to prospectively compare symptomatic improvement following a single injection of intramuscular dexamethasone (IMD) to a 3-day regimen of oral prednisone (OP) for children with mild to moderate wheezing episodes that are responsive to nebulized medications in the Pediatric Emergency Department (PED). The following children presenting with acute exacerbations of asthma to the PED were eligible for enrollment: age 3 16 years; more than two prior wheezing episodes; mild to moderate wheezing; and oxygen saturation 95% or more in room air. The study patients were randomly assigned to receive either IMD (n = 21) or OP (n = 21) in addition to a standardized treatment regimen of nebulized albuterol. All of the children were clinically rated for wheezing severity by the Pulmonary Index (PI) score at regular intervals during the study. Discharge home was based on clinical improvement during treatment in the PED; patients who were admitted to the hospital were removed from the study. Follow-up was conducted the fifth day after discharge from the ED either by clinic visit or by telephone. Patients were assessed for symptomatic improvement and relapse or clinical deterioration during the study period by a clinician blinded to group assignment. Forty-two children participated in this pilot study. There were no significant differences between the IMD and OP groups for gender or age. Mean ages were: 82 months (SD 46 months), IMD group; 63 months (SD 36 months), OP group. Clinical progress (based on PI) with treatment in the PED was the same in both groups: pretreatment median, PI = 6; PED discharge median, PI = 2. None of the study patients were hospitalized during the follow-up period, and all reported symptomatic improvement since initial treatment. The data of this pilot study suggest that IMD may be a feasible alternative to OP for treatment of acute wheezing episodes in children with asthma. IMD provides sufficient treatment to prevent clinical deterioration within 5 days after initial therapy for mild to moderate pediatric exacerbations of asthma that are responsive to nebulized medications. PMID- 9350162 TI - Will the decline in mortality from coronary heart disease in Sweden continue? PMID- 9350163 TI - Erythromelalgia: a clinical study of 87 cases. AB - We report on aetiological factors, clinical findings and prognosis of 87 patients with erythromelalgia (EM). This is the largest material reported in the western literature. There is a 100% follow up of patients with observation period up to 11 years. There were 61 females and 26 males. About two-thirds of the patients were primary cases and around three-quarters had a chronic condition. The condition was more common in lower than in upper extremities. Over time patients with erythromelalgic syndrome gradually get worse, those with primary and secondary acute EM get better, whilst primary and secondary chronic EM remain stable. PMID- 9350164 TI - Risk factors for cardiovascular disease during the period 1985-1995 in Goteborg, Sweden. The GOT-MONICA Project. AB - AIMS: To report levels of cardiovascular risk factors in 1985, 1990 and 1995 in three population samples in Goteborg, Sweden, and to compare with previous population risk factor levels. POPULATION: The study was performed within the framework of the WHO MONICA Project which compares risk factor levels as well as the incidence of coronary heart disease and stroke in 38 populations. METHODS: Three random samples of men and women aged 25-34, 35-44, 45-54 and 55-64 comprising 152-218 subjects in each age group who responded to the invitation for screening procedures which included questionnaires, physical and laboratory investigations in 1985, 1990 and 1995. RESULTS: More men than women had smoked, except for those aged 35-44 where there was no difference between men and women. The proportion of men who had smoked decreased strongly between the first and third investigations (P < 0.0001), particularly amongst the younger age-groups, with a similar tendency amongst women. In the 25-44-years age group there was a tendency towards more women than men to be smokers in 1995. Snuff was used by 27% and 19% of men aged 25-34 and 35-44 years, respectively, in 1995. Up to 5% of women used snuff; higher in the younger age groups. More young men than women reported regular physical activity during leisure time with a tendency towards an increase from 1985 to 1995. The proportion of men reporting psychological stress varied little over the study period, but women aged 25-34 reported increased stress from 1985 to 1995. Body weight increased whereas height remained stable and consequently body mass index increased in men and women (P = 0.0001). Similarly, waist:hip ratio (measured in 1990 and 1995 only) also increased (P = 0.0001). Mean systolic and diastolic blood pressure increased with age and there was also a small increase between 1985 and 1995. Systolic blood pressure increased by a mean of 1.24 mmHg per 5-year period independent of sex and age (P = 0.0001). Antihypertensive treatment increased with age, but was stable between 1985 and 1995. Serum total and LDL cholesterol concentrations increased with age, and there was a nonsignificant tendency also to higher HDL cholesterol concentrations at older ages. Serum total cholesterol concentration declined between 1985 and 1995, and HDL cholesterol declined significantly between 1985 and 1995 in all age groups for men and women only when all age groups were analysed together. Similar to total cholesterol, levels of LDL cholesterol declined between 1985 and 1995 for all ages. Serum triglyceride levels increased for men and women between 1985 and 1995. PMID- 9350165 TI - Acute intermittent porphyria: prevalence of mutations in the porphobilinogen deaminase gene in blood donors in France. AB - OBJECTIVES: Acute intermittent porphyria (AIP) is an autosomal dominant disorder resulting from a 50% deficiency in porphobilinogen deaminase (PBG deaminase). The true prevalence in the general population of mutations in the PBG deaminase gene capable of causing AIP is unknown. However, it is important to identify asymptomatic carriers of AIP mutations because all are at risk to have an acute attack. DESIGN: We measured erythrocyte PBG deaminase from 3350 healthy blood donors. When a clear cut deficiency (< mean minus 2.5 SD) was found, the PBG deaminase gene was analysed by molecular biology technics. SUBJECTS: Four subjects with PBG deaminase deficiency were identified. Two had mutations in the PBG deaminase gene which are known to cause AIP. CONCLUSION: We conclude that, in France, the mutations of the PBG deaminase gene show a high prevalence in the healthy population. If only these two confirmed latent cases are used for the calculation, in France the minimal prevalence of the AIP gene is 1:1675. PMID- 9350166 TI - Moderate beer consumption and positive biochemical changes in patients with coronary atherosclerosis. AB - OBJECTIVES: The aim of this study was to evaluate the influence of moderate beer consumption on lipid metabolism and antioxidant activity in patients (pts) with coronary artery disease (CAD). SUBJECTS: Forty-eight male pts with CAD not alcohol beverages consumers were randomly divided into experimental (EG) and control (CG) groups, 24 pts each. SETTING: Rehovot University Hospital, Israel. INTERVENTION: Every patient of the EG during a period of 30 consecutive days consumed 330 ml of Maccabee beer (> 20 g of alcohol). The pts of the CG did not consume alcohol during the trial period. METHODS: A wide range of tests including total cholesterol, LDL-C, HDL-C, total tocopherol and alpha-tocopherol. RESULTS: Only in the pts of the EG were found a tendency to an increase of the level of HDL-C and a statistically significant rise in the level of total tocopherol (P < 0.025) and alpha-tocopherol (P < 0.025). CONCLUSIONS: Even a short period of moderate beer consumption leads to some favourable biochemical changes in blood of pts with CAD which are widely regarded as indicators of CAD prevention. PMID- 9350167 TI - Impact of liver transplantation on autonomic neuropathy in familial amyloidotic polyneuropathy: an evaluation by spectral analysis of heart rate variability. AB - OBJECTIVES: To evaluate the impact of liver transplantation on familial amyloidotic polyneuropathy type I (FAP) patients' autonomic neuropathy. DESIGN: An open study. SETTING: A tertiary referral centre. SUBJECTS: Twelve liver transplanted FAP patients evaluated before and one year or longer after liver transplantation. INTERVENTIONS: Spectral analysis of heart rate variability. The low-frequency band after tilting (sympathetic), and the high-frequency band in supine position (parasympathetic) were analysed, as were the pulse and blood pressure reaction to tilting. Clinical symptoms related to autonomic disturbances were recorded. MAIN OUTCOME MEASURES: Spectral band power for sympathetic and parasympathetic activity. RESULTS: No statistically significant improvements in sympathetic or parasympathetic band power after liver transplantation was found (sympathetic band power: 2.7 (2.2-3.2) before, 2.9 (2.2-3.6) after; parasympathetic 2.0 (1.6-2.4) before and 2.0 (1.7-2.3) after. A significant correlation was noted between orthostatic blood pressure reaction and sympathetic activity before transplantation, but not after the operation. A trend was noted for improved orthostatic blood pressure reaction. Symptomatic improvements in bowel function and orthostatic symptoms were reported by several patients. CONCLUSIONS: Although improvements in autonomic symptoms are reported after liver transplantation, no significant improvement is noted in sympathetic or parasympathetic spectral band power of heart rate variability. However, the follow-up period of 17 months may be too short. Further evaluation after an additional two and four years is needed. PMID- 9350168 TI - Heart failure ketosis. AB - OBJECTIVE: To assess whether blood ketone bodies are increased in congestive heart failure (CHF). METHODS: Thirteen patients with CHF and 11 cardiac patients without CHF took part in the study. Blood acetoacetate and b-hydroxybutyrate levels and the pertinent metabolic and hormonal milieu were measured during 20 h fast and after 2 h glucose infusion. RESULTS: The averaged blood ketone body and free fatty acid levels were significantly higher during the fast and also remained higher after glucose infusion in patients with CHF than in the control group. The areas under ketone body concentration time curve over the last 8 h of the fast were 3522 +/- 662 mumol L-1 h-1 (SE) and 1789 +/- 192 mumol L-1 h-1 in patients with and without CHF, respectively (P = 0.022). Circulating noradrenaline and growth hormone were higher but glucagon lower in patients with CHF than in the controls (P < 0.05 for all differences) whereas the glucose and insulin concentrations were comparable in the study groups. At the time of peak ketonaemia the glucagon-to-insulin ratio was lower in patients with CHF than in patients without CHF (P = 0.04). CONCLUSIONS: These data suggest that severe CHF is a ketosis-prone state. Augmented supply of free fatty acids for ketogenesis due to increased stress hormone-related lipolysis is one likely mechanism. PMID- 9350169 TI - Quality of life in patients with ischaemic heart disease: a prospective controlled study. AB - OBJECTIVES: To assess quality of life in patients after acute myocardial infarction (AMI), coronary artery by-pass grafting surgery (CABG) and percutaneous transluminal coronary angioplasty (PTCA) as compared with healthy controls. DESIGN: Self-administered questionnaires were completed 1 month and 1 year after the event. SETTING: Department of Cardiology, University Hospital, Malmo, Sweden; 1989-1992. SUBJECTS: 296 AMI, 99 CABG, 18 PTCA patients and 88 randomly selected healthy controls were included; 349 patients completed the entire programme. MAIN OUTCOME MEASURES: Quality of life in the dimensions of perceived general health, thoracic pain, breathlessness, feeling of arrhythmia, anxiety, depression, self-esteem, experience of social life and sex life. RESULTS: Patients differed from controls in both psychological and somatic aspects of QL after 1 month. Furthermore, 1 month after the event AMI patients experienced more anxiety (P = 0.001) than CABG patients, whilst CABG patients experienced a poorer sex life (P < 0.001) than AMI patients. One year after the event patients differed from controls primarily in somatic symptoms: no significant differences were found across patient groups. Patients who sought emergency out-patient care during the follow-up year for clinically diagnosed angina pectoris or cardiac incompensation had reported higher levels of thoracic pain (P < 0.001) and breathlessness (P < 0.001) at 1 month follow-up than patients who did not seek such care. CONCLUSIONS: Quality of life is considerably affected in patients following a cardiac event, especially during the initial recovery phase. Although substantial improvement in quality of life occurs over time, the persistence of residual distress at 1-year follow-up is a challenge for clinicians concerned with the full rehabilitation of the cardiac patient. PMID- 9350170 TI - Bradydysrhythmia-related presyncope secondary to pheochromocytoma. AB - Pheochromocytoma endures as a life-threatening disorder. In the absence of systemic hypertension, diagnosis may be difficult. We present a 46-year-old normotensive male with a history of presyncope. One of these episodes could be documented, and revealed symptomatic bradycardia suspicious of sinus node arrest. Due to hints of an elevated sympathetic tone (Schellong test, circadian blood pressure pattern without diurnal rhythm) 24-h urinary catecholamine concentrations were measured and found increased. MIBG-scintigraphy and abdominal computed tomography indicated the location of the pheochromocytoma. After removal of the tumour, no further episodes of presyncopes or bradydysrhythmias were observed. PMID- 9350171 TI - Large phlebotomy in variegate porphyria. AB - There are no reports on effects of large blood losses in acute hepatic porphyria. In the present study we report the experiences of repeated large therapeutic phlebotomies in a patients with porphyria cutanea tarda coexisting with variegate porphyria. Neither a series of 12 phlebotomies, 300 mL each, resulting in a 17% decrease in blood haemoglobin, nor a single 400 mL phlebotomy activated the acute porphyric condition. It is concluded that the increased bone marrow metabolic throughput resulting from blood loss, in acute types of porphyria does not overload the normoblast or leukocyte precursor haem synthetic pathways in a way which will increase porphyrin precursor excretion or trigger acute porphyric symptoms. PMID- 9350172 TI - A superfemale with primary Sjogren's syndrome which involved systemic organs. AB - A 52-year-old Japanese woman complicated by a sex chromosomal anomaly as a superfemale, a mosaic of XXXXX/XXXX/XXX/XX/XO, with mild mental retardation, was hospitalized for dry mouth, dry eyes, and proteinuria. The sialography of the right parotid gland showed a globular-type gland enlargement. A definite diagnosis of primary Sjogren's syndrome (SS) was made, and further examinations revealed not only typical sicca syndrome but also systemic extraglandular lymphocytic infiltration; interstitial pneumonitis, glomerular- and interstitial nephritis, superficial gastritis, thyroiditis, and a severe excitation conductive impairment of heart. We report a very rare case of superfemale with primary SS which involved systemic organs. PMID- 9350173 TI - An unusual case of severe combined immunodeficiency with hypereosinophilia. AB - Investigation of the cytokine profile in a 26-year-old man, suffering from combined immunodeficiency with hypereosinophilia, revealed high levels of interleukin-4 and interleukin-5 and relatively low levels of interleukin-2 and interferon gamma, consistent with a T-helper type 2 pattern, as has been reported in Omenn's syndrome. However, some distinct clinical and immunological features suggest that this case may represent a unique disease with specific pathogenesis. PMID- 9350174 TI - Rapid dipstick urinalysis in the internal medicine clinic: what is missed? PMID- 9350175 TI - Resistance to activated protein C caused by the R506Q mutation in the gene for factor V is a common risk factor for venous thrombosis. AB - The protein C system is an important natural anticoagulant pathway. Protein C is the key component of the system and it is activated by thrombin bound to thrombomodulin on the surface of endothelial cells. Activated protein C (APC) inhibits coagulation by cleaving and inactivating coagulation factors factor Va and factor VIIIa. Until recently, the major genetic causes of familial venous thrombophilia were inherited deficiencies of protein C, protein S or antithrombin, but together they were found in less than 5-10% of patients with thrombosis. In 1993, the situation changed drastically with the description of inherited APC-resistance as a novel risk factor for venous thrombosis. APC resistance is characterized by a poor anticoagulant response to APC. Inherited APC-resistance is the most common genetic risk factor for this disease and it is found in 20-60% of patients. The condition is caused by a single point mutation in the gene for factor V which predicts substitution of arginine (R) at position 506 with a glutamine (Q). Mutated factor V (FVR506Q, FV:Q506 or FV Leiden) expresses normal procoagulant properties but is partially resistant to APC. The resulting hypercoagulable state confers a life long increased risk of venous but not arterial thrombosis. The FVR 506Q mutation is common in Caucasians with a prevalence of 1-15%, whereas it is not found in other human races. The FVR 506Q mutation may, due to its high prevalence, be an additional risk factor in individuals carrying other inherited defects such as deficiency of protein S, protein C or antithrombin. Such individuals have a high incidence of thrombosis and severe thrombophilia is a multigenetic disease. The high prevalence of inherited APC-resistance and the availability of easy functional and genetic tests will stimulate the development of prophylactic regimens and hopefully result in a decreased incidence of thrombosis. PMID- 9350176 TI - Familial thrombophilia: genetic risk factors and management. AB - There are now a number of potential candidates for inherited thrombophilia but a definite causal relationship has been established for only a proportion of these. Accepted causes of familial thrombophilia include the factor V Leiden defect and the prothrombin 20210 G > A variant, as well as deficiencies of antithrombin, protein C and protein S. Together these inherited abnormalities account for 30 50% of individuals presenting with venous thromboembolism. Factor V Leiden, which is present in up to 7% of the European population, is the most common cause of familial thrombophilia. On a worldwide basis its prevalence varies greatly with ethnic origin. In common with other types of familial thrombophilia the frequency of factor V Leiden is highly dependent on the population group studied. Venous thromboembolism, present in approximately 55% of individuals with familial coagulation inhibitor deficiencies, is the predominant clinical manifestation of familial thrombophilia. There are indications that the venous thrombotic risk is somewhat less in those with factor V Leiden. The thrombotic risk is markedly increased in those with combined defects and in those who are homozygous for factor V Leiden. Risk factors for thrombosis include pregnancy, including the puerperium, surgery, oral contraceptive usage and prolonged periods of immobilization. A substantial proportion of venous thrombotic events may occur spontaneously, i.e. without an obvious precipitating event. The management of patients with familial thrombophilia comprises counselling, thromboprophylaxis and thrombosis treatment. Although the immediate treatment of an acute thrombotic event is not significantly different from that of patients without recognised abnormalities, detailed patient management is seriously hampered by a lack of appropriate clinical trials. Prospective clinical studies, designed to ascertain individual thrombotic risk and to evaluate different therapeutic strategies are urgently required. PMID- 9350177 TI - Treatment of acute myelogenous leukaemia. AB - The author presents a brief overview of current therapy in adults of various ages with acute myelogenous leukemia, including remission induction and post-remission treatment and the use of haematopoietic growth factors. PMID- 9350178 TI - New strategies for the treatment of acute promyelocytic leukaemia. AB - Acute promyelocytic leukaemia (APL) is a distinct entity of acute myeloid leukaemia characterized by blast cell morphology, severe coagulopathy and t(15;17) translocation that fuses the PML gene on chromosome 15 to the retinoic acid receptor alpha (RAR alpha) gene on chromosome 17. Past experience indicated that APL is highly sensitive to anthracycline-based chemotherapy. GIMEMA experience reported a similar complete remission (CR) rate (77% versus 69%) in APL patients treated with idarubicin alone or idarubicin plus Ara-C, respectively. At present all-trans-retinoic-acid (ATRA) represents the mainstay of APL treatment. Current available clinical trials show that combination of ATRA and anthracycline induction therapy produces approximately 90% CR rate and seems to significantly improve disease-free survival. Furthermore ATRA combined therapy reduces induction death rate since ATRA syndrome has been managed with high-dose corticosteroids. However the development of ATRA resistance could limit the use of ATRA as post-remission treatment and therefore future efforts should be addressed to the search of new retinoids with comparable clinical activity, which can overcome ATRA resistance. PMID- 9350179 TI - Treatment of childhood and adult acute lymphoblastic leukaemia. AB - In the last 30 years the treatment of acute lymphoblastic leukaemia has radically changed and intensified and has resulted in improvements in the chances of cure in children to up to 70% but in adults only 30% will achieve long-term disease free survival. Data from large therapeutic trials have determined good and poor prognostic risk factors which have been of use in planning risk-directed treatment protocols and can influence the chance of cure. However intensification of treatment has also been associated with increased toxicity and significant late effects, particularly in children. In the future it will be necessary for more international collaboration and a more uniform approach to treatment in order to achieve continued improvements in the survival from this disease. In children it will be necessary to focus efforts on improving treatment of relapsed patients: chemotherapy protocols in those with a first remission of > 36 months, or for the high-risk patients with a shorter first remission, new transplantation approaches directed towards enhancing the graft-versus-leukaemia effect are going to be of increasing importance. In adults, continued efforts will be directed towards improving first remission rates with the use of increasingly intensive chemotherapeutic protocols and growth factors. The use of unrelated donor transplantation is also likely to increase, particularly in patients with 'poor risk' disease. PMID- 9350180 TI - Iron chelation therapy. AB - Desferrioxamine (DFX) remains the most effective and safe iron chelator for treatment of patients with transfusional iron overload. It is usually given by intermittent subcutaneous infusions for 8-12 h on 4-6 days weekly using a battery driven pump. Disposable balloon infusers provide a suitable method of giving continuous subcutaneous infusions with improved patient compliance. For patients with cardiac abnormalities due to iron overload, continuous intravenous desferrioxamine is essential to eliminate toxic plasma non-transferrin bound iron and to reduce body iron stores. Deferiprone (L1, l-2 dimethyl-3hydroxy-pyrid-4 one) is a less effective iron chelator but has the advantage of being orally active. Long-term trials in which patients have taken 75 mg/kg/day have shown that deferiprone is capable of maintaining body iron stores at safe levels in a proportion of thalassaemia major patients but body iron stores, assessed by liver biopsy remain at high levels (> 15.0 mg/g dry weight) in a substantial number of patients. These concentrations have been associated with tissue damage. Trials of increased doses of deferiprone (up to 100 mg/kg/day) or of combined therapy with daily deferiprone and DFX or 1 or 2 days each week are being carried out in an attempt to achieve lower body iron burden in these patients. Preliminary results show that the drugs can be given safely together and urine iron excretion produced is additive, implying that the drugs chelate different body iron pools. Patients previously well chelated with serum ferritin levels less than 2500 micrograms/L have the fewest side-effects from deferiprone and usually may be kept at the same level of body iron for periods of at least 4 years, assessed by serum ferritin and urine iron excretion. The side-effects of deferiprone result in some patients discontinuing therapy. These side-effects, especially arthropathy, mainly occur in previously poorly chelated and so the most heavily iron-loaded patients. Nausea and other gastrointestinal symptoms, agranulocytosis or milder degrees of neutropenia account with arthropathy for nearly all the withdrawals from deferiprone therapy. Patients with cardiomyopathy due to iron overload should be given intravenous DFX rather than deferiprone. Deferiprone, licensed for pharmaceutical use in India, awaits official approval for widespread clinical use in Western Europe and North America. Meanwhile, attempts to find new orally active iron chelators and improved methods of administration of desferrioxamine are in progress. PMID- 9350181 TI - New therapies for the haemoglobinopathies. AB - Re-activation of the fetal globin genes is the most realistic approach to correct the deranged pathophysiology of the haemoglobinopathies because the presence of gamma-chains can neutralize the toxic effects of the unbound alpha-globin chains in the beta-thalassaemias and inhibit the polymerization of Hb S in the sickle cell syndromes. Re-induction of fetal haemoglobin synthesis can be brought about either by direct activation of the respective promoter genes and possibly other positively acting elements, or by recruitment into proliferation and differentiation of a population of erythroid precursors which retain the gamma chain synthesis programme but remain dormant in the bone marrow of the adult unless called up in cases of acute erythroid expansion. Examples of the first group include the butyric acid and derivatives and 5' azacytidine. The second group comprises erythropoietin and a series of cytostatics, with hydroxyurea as the main representative. The activity of most of the above agents has already been studied in cell cultures and animals and confirmed in several patients, both at the haematological and biochemical level as well as through their frank clinical improvement. However, application of these drugs at large is not yet justified because a series of questions concerning their long-term efficacy, the correct dosage and timing, their tolerance and toxicity, and the potential long term dangers, including mutagenicity are still unresolved. PMID- 9350182 TI - Bone marrow transplantation for thalassaemia. AB - For all patients with a histocompatibility antigen (HLA) identical donor we are actually using two protocols to whom the patients is assigned. This is based on which class the patients belongs to at the time of bone-marrow transplant and is independent from the patient's age. For 116 patients in Class 1 and for 271 patients in Class 2 prepared for the transplant with busulfan 14 mg/kg, cyclophosphamide 200 mg/kg and cyclosporin alone, the probabilities of survival and of event-free survival are 95% and 90% for Class 1 and 85% and 81% for Class 2. For 125 Class 3 patients prepared for the transplant with busulfan 14 mg/kg, cyclophosphamide reduced to 120-160 mg/kg, cyclosporin and 'short' methotrexate, the probabilities of survival and of event-free survival are 78% and 54%. For 108 adult patients aged between 17 and 35 years, who underwent the transplant after preparation with the same protocol used for the Class 2 or Class 3 patients, the probabilities of survival are 67% and of event-free survival are 63%. Bone marrow transplantation remains the only form of radical treatment of thalassaemia in those patients with an HLA identical donor. PMID- 9350183 TI - Malignant lymphoma: current aspects in biology and classification. PMID- 9350184 TI - Treatment of follicular follicle centre lymphomas: current status and future perspectives. AB - Follicle centre lymphomas (FCLs) comprise the predominant subtype of indolent nodal lymphomas. Therapy is based on the stage of the disease and consists of extended field or total nodal irradiation in stages I and II. Patients with advanced stages III and IV may initially remain untreated and be watched until the occurrence of disease-related symptoms such as B-symptoms, haematopoietic insufficiency, lymphoma progression or bulky disease. On the occurrence of these signs a cytoreductive chemotherapy of mild to moderate intensity such as cyclophosphamide, vincristine, prednisone (COP) or mitoxantrone, chlorambucil, prednisone (MCP) should be initiated. In responding cases maintenance with interferon-alpha (IFN alpha) leads to a significant prolongation of the progression-free interval. Modifications of this approach include the upfront combination of IFN alpha with anthracycline containing combinations such as cyclophosphamide, doxorubicin, teniposide, prednisone (CHVP). New perspectives arise from the introduction of myelo-ablative radio-chemotherapy with subsequent stem-cell transplantation and antibody-based immunobiological therapies. PMID- 9350185 TI - Treatment of B-cell chronic lymphocytic leukaemia: current status and future perspectives. AB - In the last two decades, important advances have been made in the biology, natural history, and prognosis of B-cell chronic lymphocytic leukaemia (CLL). In addition, treatment possibilities for patients with CLL have changed as a result of the identification of prognostic factors for survival and the availability of new drugs and treatment strategies. Patients in the early clinical stages (Binet A, Rai 0) with stable disease have a probability of long survival and should not be treated unless the disease progresses. In contrast, most patients with poor prognostic features, such as an advanced clinical stage (Binet B, C; Rai III, IV), diffuse bone-marrow infiltration or rapidly increasing blood lymphocyte levels, have a median survival probability of < 5 years and require therapy. Purine analogues are highly effective. Among these, fludarabine has become the treatment of choice for patients failing standard therapies. The role of purine analogues either alone or in combination with other drugs as front-line therapy is being investigated. Certain situations (e.g. autoimmune cytopenias, hypersplenism) require special treatment approaches (e.g. corticosteroids, splenectomy). Transplants of progenitor haematopoietic cells are also increasingly performed and deserve further investigation in younger patients with poor prognostic features. As a result of these advances, symptoms palliation is no longer the only possible goal in CLL therapy; sustained remissions and even cures are likely to be obtained in the near future. PMID- 9350186 TI - Empirical and subsequent use of antibacterial agents in the febrile neutropenic patient. AB - The objective of this analysis were an assessment of the feasibility of a more individually tailored approach of empirical antibiotic therapy in febrile neutropenia and an exploration of the reasons to modify the initial regimen. DESIGN, SETTING AND SUBJECTS: The main source was a database on febrile neutropenic cancer patients from an unblinded large trial conducted in 35 centres world-wide. This was supplemented by data from patients enrolled in a consecutive series of randomized trials at the Department of Haematology, University Hospital Nijmegen. INTERVENTIONS: Diagnostic procedures were standardized, types of possible infections defined and the reasons for modifying an empirical regimen were recorded. MAIN OUTCOME MEASURES: Survival of the febrile neutropenic episode, development of microbiologically and clinically defined infection in relation to causative organisms, and results of modification. RESULTS: Monotherapy was as effective as combination therapy with an overall mortality of < or = 7%, with 21% of neutropenic episodes accompanied by a clinically defined infection proving fatal compared with only 4% of episodes without a focus. At the end of treatment the empirical regimen had been added to in 60% of cases in the multicentre trial, in contrast to 39% in our own institution, in many cases simply because of continuing fever. CONCLUSION: The development of local guidelines for individually tailoring antibiotic therapy by complementing the empirical regimen is a feasible option for achieving an optimal anti-infective strategy for febrile neutropenic cancer patients. PMID- 9350187 TI - Is there a rationale for the use of antimicrobial prophylaxis in neutropenic patients? AB - Antimicrobial prophylaxis in neutropenic patients has been practised in one form or another for several decades but the goal is no longer clear. From being initially solely an attempt at decontamination, drugs such as co-trimoxazole and later the fluoroquinolones were preferred to non-absorbable regimens because they achieve reliable protection against bacteraemia due to Gram-negative bacilli. Nevertheless, fever still invariably occurs during neutropenia leading to the initiation of traditional empirical therapy. Not only is this approach illogical but it also ignores the flexibility afforded the oral and parenteral formulations of the fluoroquinolones. Instead, it might be as effective and less costly if these agents were given orally until the end of neutropenia unless there was evidence of malabsorption or poor oral intake, in which case treatment would be continued parenterally. Should patients develop fever, an attempt would be made to complement treatment with another anti-microbial agent for microbiologically or clinically defined infection. This would be carried out at diagnosis, before any changes in the prophylactic regimen could be made. Otherwise, treatment with the prophylactic regimen would continue without modification. There is a less compelling need for prophylaxis against candidosis, herpes simplex and cytomegalovirus disease as these would be better managed pre-emptively when there is evidence of yeast carriage or re-activation of viral infection. Similarly, prophylaxis of aspergillosis is a forlorn hope and again a pre-emptive approach might serve us better once there is a screening test available and a safe and effective drug. PMID- 9350188 TI - Antifungal treatment in patients with cancer. AB - Invasive fungal infections are one of the leading causes of morbidity and mortality in cancer patients. Amphotericin B deoxycholate is still considered the gold standard of antifungal therapy, although the new triazoles (itraconazole and, especially, fluconazole) have shown to be able to replace amphotericin B for some therapeutic indications. The new lipid formulations of amphotericin B have disclosed new therapeutic perspectives, especially in patients with severe renal failure and documented, infections. At this time, indications, contraindications and limitation of the various drugs in the antifungal armamentarium are still partially unclear. Antifungal prophylaxis with fluconazole may be indicated in high-risk patients, although the duration of such prophylaxis should be limited as much as possible, in order to prevent selection of resistant strains and acquired resistance. Empirical antifungal therapy is used extremely widely (maybe, too widely) in many cancer centres, despite being based on limited clinical data. For this indication, fluconazole may also be effective in patients not receiving fluconazole prophylaxis, in whom Aspergillus infection is unlikely. PMID- 9350189 TI - Gene therapy of haematopoietic cells. AB - Gene therapy of haematopoietic stem cells (HSCs) has been under investigation for 15 years. HSCs can be easily transduced with retroviral vectors (Moloney murine leukaemia virus = MMLV) in the mouse and expression of transferred genes can be achieved in long-term reconstituted mice. While human haematopoietic progenitor cells can be transduced with high efficiency using amphotropic MMLV retroviral vectors, and expression of the transferred gene is easily obtained in their progeny cells, it has proven problematic to transfer genes efficiently into the HSCs of humans in clinical trials. Efforts are now under way, in many laboratories, to increase the gene transfer efficiency of retroviral vectors, or alternatively, to develop new vectors that can transduce quiescent human HSCs with higher efficiency than is currently possible. This brief review will address the two main research areas that are being explored. Firstly, investigations in human haematopoiesis to gain understanding into the molecular and cellular mechanisms that control HSC behaviour in vitro and in vivo. Secondly, development of new vectors that can transfer genes to quiescent human HSCs. PMID- 9350190 TI - Gene transfer trials in clinical haematology. AB - The haematopoietic stem cells in the bone marrow have long been considered, at least in theory, as an ideal target for gene therapy. Of the more than 250 clinical gene transfer protocols reported from around the world up to December 1996, almost one in three involves manipulation of haematopoietic cells. This includes both marketing trials and trials with a therapeutic intent. The human gene transfer trials targeting haematopoietic cells, the knowledge gained from them and their limitations are briefly summarized. PMID- 9350191 TI - Current challenges in cancer gene therapy. AB - The ability to transfer novel genes into mammalian cells has allowed us to conceive of novel strategies towards cancer therapy. Since the initial gene transfer clinical trial in 1989, over 300 cancer patients have been enrolled in gene therapy trials. Despite this, an NIH-sponsored panel concluded that 'clinical efficacy has not been definitively demonstrated at this time in any gene therapy protocol'. However, the first 8 years of gene therapy research have provided us with insights regarding the areas that require further scientific progress. For example, it is now clear that while in vitro assays of gene modified haemotopoietic progenitor cells suggest high transduction efficiencies, once these cells are infused in vivo, only a small percentage of circulating transduced cells can be detected. While the initial clinical studies have demonstrated that gene transfer in patients can be safe and feasible, they have also indicated that future research is necessary towards the development of improved gene transfer techniques for these approaches to be successful. PMID- 9350192 TI - Understanding von Willebrand's disease from gene defects to the patients. AB - von Willebrand's disease (vWD) is caused by qualitative (type 2) and quantitative (types 1 and 3) abnormalities of von Willebrand factor (vWF). vWD type 3, a severe form of the disease with nearly complete deficiency of the protein in plasma, are found to be homozygous or compound heterozygous for null mutations in the vWF gene. Null mutations in both alleles of the vWF gene completely disrupt the protein synthesis resulting in a nearly complete deficiency of the vWF in the type 3 patients. The vWD type 1 patients (mild form with partial deficiency of the protein) could be heterozygous for null mutations or compound heterozygous for the mutations (null mutation + missense mutation) in the gene. The vWD type 2, divided into four variants: types 2A, 2B, 2M and 2N, are caused exclusively by missense mutations within three different domains of the protein (gain or loss of function). The majority of type 2A mutations are located in the A2 domain and the types 2B and 2M mutations are in the A1 domain, while the type 2N mutation is in the FVIII binding domain. PMID- 9350193 TI - The problem of diagnosing von Willebrand's disease. AB - Diagnosis of von Willebrand's disease (vWD), particularly vWD Type 1, remains a clinical problem for several aspects. Its definitive diagnosis requires documentation of three factors: bleeding, low levels of qualitatively normal von Willebrand factor (vWF), and inheritance. In the absence of any of these factors the diagnosis may be only merely 'possible', or even unacceptable. Laboratory diagnosis of vWD includes screening tests and confirmatory tests. vWD Types 2 and 3 are relatively easy to diagnose and appear to be genetic disease of a single locus, the vWF gene. As new genetic and possibly non-genetic factors are discovered, the diagnosis of vWD Type 1 may become easier. PMID- 9350194 TI - Treatment of von Willebrand's disease. AB - von Willebrand's disease is the most frequent of inherited bleeding disorders (1:100 affected individuals in the general population). The aim of therapy is to correct the dual defects of haemostasis, i.e. abnormal coagulation expressed by low levels of factor VIII and abnormal platelet adhesion expressed by a prolonged bleeding time. There are two main options available for the management of von Willebrand's disease: desmopressin and transfusion therapy with blood products. Desmopressin is the treatment of choice in patients with Type 1 von Willebrand's disease, who account for approximately 80% of cases. The pharmacological compound raises endogenous factor VIII and von Willebrand factor and corrects the prolonged bleeding time in most patients. In Type 3 and in the majority of Type 2 patients, desmopressin is not effective and it is necessary to resort to plasma concentrates containing factor VIII and von Willebrand factor. Treated with virucidal methods, these concentrates are currently effective and quite safe, even though the bleeding time defect is not always corrected by them. Platelet concentrates or desmopressin can be used as adjunctive treatments when poor correction of the bleeding time is associated with continued bleeding. PMID- 9350195 TI - Transport proteins in drug resistance: biology and approaches to circumvention. AB - At least two transport proteins, P-glycoprotein (Pgp) and the multidrug resistance associated protein (MRP), are believed to play a significant role in clinical resistance to cytotoxic therapy. These proteins are expressed at relatively high levels in a number of malignant diseases including various types of leukaemias. They are variably expressed on both the plasma membrane and intracellular vesicular membranes resulting in cellular drug efflux or vesicular drug sequestration, respectively. The action of MRP as a drug transporter depends on intracellular levels of glutathione. A number of strategies for circumvention of these drug resistance mechanisms have been developed and some of these are now in clinical trial. PMID- 9350196 TI - Avoidance of apoptosis as a mechanism of drug resistance. AB - Inherent or acquired drug resistance is a major obstacle for the successful treatment of cancers. Many mechanisms of drug resistance have been described including a decreased drug uptake, an increase in DNA damage repair, enhanced drug detoxification, an altered level or mutation of the intracellular drug target or an increased drug efflux from the cell. Most of these mechanisms impinge upon the interaction of a drug with its cellular target or immediate consequences of such as interaction. For example, a decrease in the cellular levels of topoisomerase II thwarts the efficacy of certain topoisomerase II inhibitors, and enhanced levels of glutathione increase resistance to DNA alkylating agents. However, some tumours are inherently resistant to all chemotherapeutic agents, i.e. with different mechanisms of action. What is the mechanism(s) underlying this pleiotropic drug resistance? One possibility is that such drug-resistant tumour cells have an abnormally high threshold for the engagement of apoptosis (programmed cell death). The suppression of apoptosis as a mechanism for drug resistance is discussed in this article. PMID- 9350197 TI - Transport proteins in drug resistance: detection and prognostic significance in acute myeloid leukemia. AB - Resistance to natural product-derived anti-cancer drugs, such as the anthracyclines and etoposide, contributes to the failure of chemotherapeutic treatment of leukaemia. One biological resistance mechanism of potential importance is the overexpression of the plasma membrane drug transporter proteins P-glycoprotein (Pgp) and multidrug resistance protein (MRP). Many studies have reported evidence for a correlation of Pgp/MDR1 expression with unfavourable prognostic features in acute myeloid leukaemia (AML). Failure to achieve complete remission (CR) is correlated with Pgp and the CD34+ phenotype. For MRP fewer data are available, which suggest a basal expression level in most AMLs. Another protein reported to correlate with treatment failure in AML is the lung resistance protein or major vault protein (LRP), a protein with a still unknown function. Co-expression of Pgp and LRP especially seems to define an adverse prognostic population. Further progress towards the understanding of the clinical importance of these proteins is hampered by the lack of validation of methods to determine their expression. A reliable way to measure Pgp seems to be the assessment of the active transport of fluorescent Pgp substrates, such as rhodamine 123 out of AML cells. Such functional Pgp assays can be used to validate mRNA or protein measurements and to quantify the effect of Pgp or the magnitude of the effect of a blocker of the Pgp-mediated drug efflux on the intracellular drug concentration. The prognostic value of such methods has still to be shown. PMID- 9350198 TI - Kaposi's sarcoma and the new herpesvirus. PMID- 9350199 TI - Role of haemolytic and non-haemolytic phospholipase C from Pseudomonas aeruginosa in interleukin-8 release from human monocytes. AB - A massive accumulation of neutrophils, mainly due to enhanced interleukin-8 (IL 8) levels, is believed to contribute to the deleterious effects of Pseudomonas aeruginosa lung infection, e.g., in cystic fibrosis (CF). Antibodies to phospholipase C, an exoenzyme of P. aeruginosa, are detected early and at high levels in CF patients. However, P. aeruginosa produces at least two types of phospholipase C (PLC), one haemolytic (PLC-H) and the other non-haemolytic (PLC N), both with mol.wts of c. 77 kDa. Experiments were performed to evaluate the potential contribution of P. aeruginosa PLC to neutrophil accumulation during infection. Therefore, P. aeruginosa PLC-H and PLC-N were compared with regard to IL-8 generation from human monocytes. Purified PLC-H as well as culture supernates (mol.wt > 50 kDa) of a P. aeruginosa strain capable of producing both PLC-H and PLC-N, and mutant strains deficient in the production of one or other phospholipase, or both, were examined. Purified PLC-H (only at low concentrations up to 1 unit/4 x 10(5) monocytes), induced a dose-dependent increase in IL-8 release and IL-8-specific mRNA expression over that of unstimulated cells (at 4-, 12- and 24-h incubation times). Higher concentrations of PLC-H led to a decrease in IL-8 release and IL-8-specific mRNA expression. These findings were confirmed by the results obtained with the supernates of cultures of mutant strains of P. aeruginosa PAO1 that produced either a PLC-H or PLC-N or neither. Stimulation and inhibition of IL-8 release and mRNA expression were associated with a culture supernate fraction of mol. wt > 50 kDa and containing PLC-H. These results contribute to the understanding of the role of both P. aeruginosa PLC in IL-8 generation during their interaction with human monocytes. PMID- 9350200 TI - Composition of staphylococcal bi-component toxins determines pathophysiological reactions. AB - Staphylococcus aureus produces numerous bi-component toxins, e.g., Panton Valentine leukocidin (Luk-PVL) and gamma-haemolysin, which consist of type S and F proteins. Previous studies showed that Luk-PVL induces inflammatory mediator release from human granulocytes that might reflect the in-vivo effects, e.g., dermonecrosis by Luk-PVL. Clinical isolates not only harbour the two genes coding for Luk-PVL (S-protein: LukS-PVL, F-protein: LukF-PVL) but also the three genes encoding gamma-haemolysin (S-protein: HlgA, HlgB; F-protein: HlgC). The interaction of all the possible potential toxins with human granulocytes was studied with regard to the generation of oxygen metabolites (chemiluminescence response), enzyme activity (beta-glucuronidase) and histamine release as well as interleukin (IL)-8 generation. The data clearly show that the individual subunits (S, F) differ in their activities. The following activities were obtained for the S components: LukS-PVL > HlgC > HlgA; the F components LukF-PVL and HlgB were similarly active. Thus, the toxins LukS-PVL/LukF-PVL and LukS-PVL/HlgB were the most potent inducers of inflammatory mediator release from human granulocytes, followed by HlgC/LukF-PVL and HlgC/HlgB and to a lesser degree by the toxins HlgA/LukF-PVL and HlgA/HlgB. The data indicate that class S components and class F components are interchangeable and give toxins with genuine biological activities. PMID- 9350201 TI - Channel-forming leucotoxins from Staphylococcus aureus cause severe inflammatory reactions in a rabbit eye model. AB - Panton-Valentine leucocidin arises from the combination of one S component (LukS PV) with one F component (LukF-PV), whereas gamma-haemolysin comprises two S components (HlgA and HlgC) with one F component HlgB. The intravitreal injection of rabbit eye with the six combinations (S + F) of channel-forming leucotoxins produced by Staphylococcus aureus ATCC 49775 induced acute inflammatory reactions depending on time and doses of toxins. These reactions involved posterior chamber as well as anterior chamber and conjunctiva, eyelids and annexes. Histological examination confirmed the involvement of eye tissues and the disruption of the retinal barrier. The lesions began only 4 h after injections and persisted for at least 5 days. Clinical and biological effects of each leucotoxin were modulated by the speed of onset and intensity of inflammation and necrosis, leading to a functional classification according to the severity of the lesions (HlgA + LukF PV > HlgA + HlgB > or = LukS-PV + HlgB > or = LukS-PV + LukF-PV > HlgC + HlgB > or = HlgC + LukF-PV). Moreover, N-acetyl beta-D glucosaminidase assays on crude extracts of vitreous revealed granules and granule secretions from polymorphonuclear cells with levels according the above classification. These results show that channel-forming leucotoxins have a very significant inflammatory activity. As most S. aureus strains produce two or even six leucotoxins depending on the production of Panton-Valentine leucocidin, these compounds could be considered to be virulence factors. PMID- 9350202 TI - Detection of Mycobacterium tuberculosis DNA in blood of patients with acute pulmonary tuberculosis by polymerase chain reaction and non-isotopic hybridisation assay. AB - The detection of Mycobacterium tuberculosis DNA in peripheral blood mononuclear cells (PBMC) by PCR and non-isotopic hybridisation assay was evaluated for the laboratory diagnosis of pulmonary M. tuberculosis infection. The PCR technique was based on the presence of IS6110, a DNA sequence specific for M. tuberculosis, and performed on PBMC from 30 patients belonging to the fifth group of the American Thoracic Society (ATS) classification of tuberculosis. The identification of amplification products was confirmed after electrophoresis by hybridisation with a non-isotopic probe in a DNA enzyme immunoassay (DEIA). Of the 30 blood samples studied by the PCR-DEIA technique, 26 gave positive results and four gave negative results. Blood samples from 30 subjects in a control group were negative by this technique. The data suggest that PCR-DEIA of blood may provide a sensitive, specific and useful means of diagnosing mycobacterial infection. PMID- 9350203 TI - Genetic differentiation of Australian isolates of Mycobacterium tuberculosis by pulsed-field gel electrophoresis. AB - As part of an epidemiological study of tuberculosis in Australia, 84 isolates of Mycobacterium tuberculosis from patients were analysed by pulsed-field gel electrophoresis (PFGE). The isolates were genetically heterogeneous, with 66 different DNA banding patterns obtained following digestion of genomic DNA with Dra1 and 53 patterns with Xba1. When the results were compared with those previously obtained in restriction fragment length polymorphism analysis (RFLP), in 87% of cases the results with Dra1 were consistent with those obtained with insertion sequence IS6110 as a probe in RFLP. However, PFGE was able to differentiate four of eight isolates which were identical with IS6110 typing. The high polymorphism amongst strains and the high average age of the patients (51 years) suggested that most organisms were cultured from patients who had reactivation of existing infections. Isolates with identical DNA patterns were found in different states of Australia, but no one strain predominated in any area. This suggests that tuberculosis has been introduced into Australia from various sources. PMID- 9350204 TI - Pulsed-field gel electrophoresis genomic fingerprinting of hospital Escherichia coli bacteraemia isolates. AB - Pulsed-field gel electrophoresis (PFGE), because of the increased sensitivity it affords over other methods of bacterial genotyping, represents a potentially powerful tool for the characterisation of isolates from hospital infections. Genomic fingerprinting by PFGE was applied to all clinical isolates of Escherichia coli obtained from blood during a 6-month period (78 isolates, 58 patients) at the University of Michigan Medical Center. The rare-restriction patterns of these isolates, in contrast to those of isolates from the E. coli reference collection (ECOR), were not randomly distributed through the E. coli species. Four related clusters, which represented c. 21% of the blood isolates, were identified. Two of these genotypic clusters were also clustered temporally, their members all being isolated within the same 2-week period, while the other two clusters spanned the study period. These observations indicate in-hospital endemic vectors or the occurrence of specialised E. coli lineages that are capable of invading the bloodstream and exploiting in-hospital vectors, or both. PMID- 9350205 TI - International quality control of phage typing of Staphylococcus aureus. International Union of Microbial Societies Subcommittee. AB - A questionnaire was sent to the 48 national typing centres for Staphylococcus aureus and 31 replies were received. Methods of phage typing varied and molecular methods were not universally available, although pulsed-field gel electrophoresis was offered by 13 centres. Results for a quality control phage typing exercise were received from 25 centres. Increased standardisation of methods and definitions are indicated. Differences from the consensus patterns were mainly due to typing at an inappropriate dilution of phage, but five strains caused difficulties in many centres. Overall reproducibility was good. Phage typing remains a cost-effective method for epidemiological studies, particularly on a large scale. The strains selected for the quality control exercise included many strains suitable for controlling molecular methods as well as testing phage typing. Molecular methods help in the validation of the conclusions which may be drawn from phage typing. PMID- 9350206 TI - Risk factors, aetiology, therapy and outcome in 123 episodes of breakthrough bacteraemia and fungaemia during antimicrobial prophylaxis and therapy in cancer patients. AB - One hundred and twenty-three breakthrough bacteraemias (BB) were defined during a 5-year period in a National Cancer Centre, among 9986 admissions and a total of 979 bacteraemic episodes analysed. Of 123 bacteraemias in 103 patients, 77 were polymicrobial and 116 of the 323 organisms isolated were resistant to currently administered antimicrobial agents. Sixty-seven of the bacteraemic episodes were catheter-associated, as confirmed by the isolation of the same organisms from both blood and catheter tip. The strains isolated most frequently were coagulase negative staphylococci (30.5%), corynebacteria (10%), Pseudomonas aeruginosa (10%), Enterococcus faecalis (9%) and viridans streptococci (8.5%). Gram-positive aerobes accounted for two-thirds of all micro-organisms isolated during breakthrough bacteraemic and fungaemic episodes. Polymicrobial episodes were associated more frequently with vascular catheters and neutropenia, and had a less favourable outcome than monomicrobial infections. Relapse was associated more frequently with catheter-related episodes, but the overall mortality rate was similar and independent of catheter insertion. Breakthrough bacteraemic and fungaemic episodes were associated more frequently with acute leukaemia. Catheter removal, as an independent variable, and modification of antimicrobial therapy were essential for better outcome. PMID- 9350207 TI - Media for the detection and recognition of the enteropathogen Providencia alcalifaciens in faeces. AB - A medium (PAM: Providencia alcalifaciens medium) is described that enables the presence of the enteropathogen P. alcalifaciens in faeces to be detected with ease and simplicity. This organism is probably the only oxidase-negative organism likely to be present in tetrathionate broth cultures of faeces that is unable to ferment the mannitol, xylose or galactose present in the medium. Thus the red colonies of P. alcalifaciens appeared quite distinct from the lemon-yellow acid forming colonies of all the other bacteria that ferment one or more of these sugars. Extensive tests showed the medium to be both highly specific and sensitive in detecting P. alcalifaciens. Two additional media are described that enable the identity of presumptive P. alcalifaciens isolates to be confirmed unequivocally and with ease. PMID- 9350208 TI - T-cell transfer and cytokine/TCR gene deletion models in the study of inflammatory bowel disease. AB - Until recently there existed no appropriate immunological animal models for human inflammatory bowel diseases (IBD). Today a number of models, mostly in the mouse and rat, have proved useful in the study of several aspects of IBD, including the histopathology and the disease-inductive and -protective cell types, subsets and cytokines, for example CD4+ T cells, IFN gamma, IL-12, IL-2, IL-10 and TGF beta. Furthermore, these recent IBD models make it possible to examine various chemo- and immunotherapeutic approaches. This review focuses on IBD development in adoptive T-cell transfer models and in gene-deleted mice. PMID- 9350209 TI - Effects of monocyte purification and culture on integrin expression. AB - Selective alterations in the surface expression of members of the LeuCAM (leukocyte cell adhesion molecule) family of integrins occur during in vitro culture of human monocytes. Such changes may relate in part to cellular maturation, but also to activation following purification and culture of monocytes. In this paper, we examined the effects of monocyte isolation, adherence during culture and endotoxin exposure on the expression of these molecules and the ligand for LFA-1, ICAM-1 (CD54). Expressions of CD11b, CD18 and CD54, but not CD11a or CD11c, were higher on monocytes freshly isolated by density gradient separation and plastic adherence as compared with cells labelled directly in whole blood. However, the surface expression of the LeuCAMs and CD54 on cultured monocytes was not affected by short-term adherence to plastic for 2 h, as determined by comparisons of their expression on adherence-isolated and elutriated monocytes. In contrast, prolonged adhesion of monocytes for up to 21 days in culture altered expression of CD11a without affecting that of the other LeuCAMs or CD54. Expression of CD11a decreased more rapidly on adherence maintained cells as compared with suspension-cultured cells. Our results show that cellular manipulations required for in vitro studies of monocyte/macrophages may alter expression of the LeuCAMs. PMID- 9350210 TI - Modification of mononuclear cell function after incubation with albumin-bound unsaturated fatty acids or soybean oil emulsion. AB - Administration of total parenteral nutrition (TPN) with soybean oil emulsion leads to a linoleic acid enrichment of the plasma membrane that may explain an in vivo activation of mononuclear cells (MNC) seen in our previous studies. Fatty acids from the lipid emulsion may have been accessible to MNC after endocytosis of lipid particles, or by direct uptake of fatty acids after lipoprotein lipase hydrolyzation of the emulsion triglycerides. To resemble the incorporation of fatty acids in vivo, we have modified MNC membrane lipid composition by incubation with different albumin-bound unsaturated fatty acids (UFA) or soybean oil emulsion. After incubation with albumin-bound linoleic and oleic acid, the unstimulated release of superoxide anion was unchanged, while zymosan-stimulated release was 140% (n.s) and 112% (p < 0.05) and phorbol-myristate-acetate (PMA) stimulated release 148% (p < 0.05) and 124% (p < 0.05) of controls, respectively. Incubation with other UFAs or emulsion did not change superoxide anion release. Unstimulated lymphocyte proliferation increased 3 to 13-fold (p < 0.05) after incubation with all UFAs compared to controls, while UFA incubation did not change phytohemagglutinin (PHA) or PMA-stimulated proliferation. Unstimulated lymphocyte proliferation was decreased after incubation with emulsion, while PHA/PMA-stimulated proliferation was unchanged. Increase in membrane fluidity was detectable only after incubation with emulsion. The increased reactivity may have been caused by changes in the lipid environment surrounding membrane-bound enzymes important for signal transduction through the plasma membrane. PMID- 9350211 TI - Characterization of the lactoferrin-dependent inhibition of the adhesion of Actinobacillus actinomycetemcomitans, Prevotella intermedia and Prevotella nigrescens to fibroblasts and to a reconstituted basement membrane. AB - Lactoferrin was previously shown to inhibit the adhesion of A. actinomycetemcomitans, P. intermedia and P. nigrescens to human cells. Lactoferrin was also shown to competitively inhibit the binding of these bacteria to the basement membrane protein laminin. The present study aimed to determine the type of interactions inhibited by lactoferrin. Lactoferrin binds to fibroblast monolayers and Matrigel, a reconstituted basement membrane, through ionic interactions. The adhesion of A. actinomycetemcomitans to these substrata was mainly dependent on the ionic strength of the environment. P. intermedia and P. nigrescens also adhere to fibroblasts mainly by ionic interactions, while their adhesion to Matrigel seems to be mediated by specific mechanisms. Lectin type interactions were not found to be involved in the binding of these bacteria to the substrata. Treatment of either A. actinomycetemcomitans or fibroblasts with lactoferrin decreased the adhesion in a dose-dependent manner, while lactoferrin treatment of Matrigel alone had no adhesion-counteracting effect. Adhesion of P. intermedia and P. nigrescens to Matrigel was not significantly affected by the ionic strength, but the presence of lactoferrin inhibited the adhesion. Lactoferrin bound to Matrigel, P. intermedia and P. nigrescens was rapidly released, while lactoferrin bound to A. actinomycetemcomitans and fibroblasts was retained. These findings indicate that lactoferrin-dependent inhibition of the adhesion of A. actinomycetemcomitans, P. intermedia and P. nigrescens to fibroblasts and Matrigel can involve binding of lactoferrin to both the bacteria and substrata. The decreased adhesion may be due to blocking of both specific adhesin-ligand as well as non-specific charge-dependent interactions. PMID- 9350212 TI - A short exposure to a high-glucose milieu stabilizes the acidic vacuolar apparatus of insulinoma cells in culture to ensuing oxidative stress. AB - It was recently suggested that extracellular hydrogen peroxide, after diffusing into and throughout adjacent cells--which may be the case if they have only a weak capacity to degrade hydrogen peroxide--labilizes their lysosomal compartment due to its content of low-molecular-weight iron in redox-active form. The iron would be present as a consequence of normal autophagocytotic degradation of various iron-containing metalloproteins. Beta- and insulinoma cells are especially vulnerable to oxidative stress, since they possess only low capacity to degrade hydrogen peroxide, and, perhaps, since they normally have a certain degree of autophagocytotic degradation of secretory granules with some iron content--crinophagy. The toxicity to beta cells of oxidative stress, such as an exposure to alloxan, that results in extracellular formation of hydrogen peroxide, is considerably reduced if animals are initially given an intravenous bolus dose of glucose, temporarily bringing up the blood level to about 20 mM. In this study it was demonstrated that already as short an exposure as 30 min to 20 mM D-glucose reduces the sensitivity of HIT and NIT insulinoma cells in culture to a subsequent exposure to hydrogen peroxide. In parallel, exposure to such a high-glucose medium also reduces their desferrioxamine-available amount of iron and, moreover, stabilizes their lysosomal membranes against oxidative stress- thus preventing diffusion to the cytosol of damaging lysosomal contents following iron-catalyzed, Fenton-type, intralysosomal reactions. We suggest that both general autophagocytotic turnover and, in particular, crinophagy of secretory granules are decreased by an increased glucose concentration of the surrounding milieu, with attendant reduced amounts of intralysosomal low-molecular-weight iron and, thus, diminished sensitivity to oxidative stress. PMID- 9350213 TI - Inhibition of phytohaemagglutinin-induced lymphoproliferation by soluble annexin II in sera from patients with renal cell carcinoma. AB - Annexin II (AII) is a member of a family of glycoproteins which bind negatively charged phospholipids in a calcium-dependent manner. Annexins are membrane associated proteins, expressed both in normal and malignant cells, but have also been detected as soluble molecules in serum and other body fluids. Because of their adhesive properties, it has been suggested that annexins play a role in the metastatic process. An ELISA was established for quantification of soluble AII. Within-run variation was 5.2-10.4% and run-to-run variation 12.4-15.6%. Soluble AII was detected in all sera studies. A strongly positive serum was arbitrarily given the value 100 AII units and used as reference serum. The mean level in sera from 20 normal blood donors was 49 (SE 5.6) AII units. Sera from peripheral blood of five patients with renal cell carcinoma and sera from blood obtained from the renal vein of the same patients contained 47 (SE 20) and 83 (SE 28) AII units, respectively. In two patients, AII levels were increased in renal vein serum as compared with peripheral blood serum. Interestingly, in both cases, and in none of the three remaining cases, phytohaemagglutinin-stimulated lymphoproliferation was suppressed by renal vein serum as compared with peripheral blood serum. Affinity absorption of AII from the renal vein sera with increased AII levels strongly reduced their immunosuppressive activity. Addition of affinity-purified AII to cell cultures suppressed lymphoproliferation. These data show that the level of AII is markedly increased in renal vein sera from some patients with renal cell carcinoma, suggesting that AII may be locally released in vivo. The study also demonstrates an immunosuppressive effect of soluble AII in vitro. We speculate that soluble AII released by the tumour has immunosuppressive properties. This study identifies soluble AII as a novel immunosuppressive factor in sera from patients with renal cell carcinoma. A further study including a larger number of patients is currently in progress, in order to investigate the pathological significance of this finding. PMID- 9350214 TI - Purulent meningitis due to Rhodococcus equi. A case of posttraumatic infection. AB - Opportunistic infections due to Rhodococcus equi have been increasingly reported in the immunocompromised population, especially in patients with AIDS. In this report, we present an unusual case of purulent meningitis that developed in an immunocompetent six-year-old child through direct inoculation of R. equi. PMID- 9350215 TI - Cefuroxime resistance in Klebsiella pneumoniae. Susceptibility to cefotaxime and ceftazidime despite production of ESBLs. AB - The production of beta-lactamases, the outer membrane protein (OMP) patterns, some clinical impacts and the prevalence of resistance among cefuroxime-resistant Danish clinical isolates of Klebsiella pneumoniae were investigated. Fifteen resistant and five susceptible strains were collected from 14 patients during 1991-1994. Isolates from five patients produced extended-spectrum beta-lactamases (ESBLs). Cefuroxime resistance was accompanied by a 10-fold elevation of ciprofloxacin minimal inhibitory concentration (MIC), and for some isolates by an alteration of the OMP pattern. The relationship between alterations of the OMP patterns and cross-resistance to ciprofloxacin and the other antibiotics tested was not universal. Ten of the cefuroxime-resistant strains had elevated MICs of cefotaxime or ceftazidime, but the MICs were still below the breakpoint for susceptibility. The MICs of imipenem were not affected. Nosocomial infection or long-term colonization with resistant strains may be of importance since five patients were not treated with cefuroxime prior to isolation of the resistant strain, and all patients had either serious diseases or stayed at the hospital for a long period of time. The prevalence of cefuroxime and ciprofloxacin resistance among clinical isolates from Copenhagen county during 1990-1995 was 8.3% and 7.5%, respectively, but higher for urinary tract specimens. A greater consumption of cefuroxime as compared to cefotaxime and ceftazidime in this study, as seen generally in Denmark, indicated that ESBLs produced by the investigated strains of K. pneumoniae may be selected with cefuroxime. PMID- 9350216 TI - Sulphasalazine, olsalazine and sulphapyridine induce mitogenic actions in the rat intestinal epithelium. AB - Our aim was to study the influence of sulphasalazine (SASP), olsalazine (ADS) and sulphapyridine (SP) on the cell kinetics of the intestinal epithelium in conventional rats. Groups of rats were treated with SASP, ADS or SP for 9 days. After an intraperitoneal injection of a metaphase blocker, the rats were killed and the jejunum, ileum and colon were examined in histological sections by means of the cumulative mitotic index (MI), growth fraction and number of cells in crypts and villi. SP increased both the MI in the jejunum, ileum and colon and the number of crypt cells (p < 0.05 vs controls). In contrast, SASP and ADS increased the MI only in the colonic epithelium (p < 0.05 vs controls). The growth fraction was essentially unaffected. Our results suggest that SASP, SP and ADS have a selective compartment-dependent proliferative action on the epithelium of the intestinal tract. PMID- 9350217 TI - Transduction potential of human retroviruses in highly proliferating small-cell lung cancer cells as well as non-proliferating hematopoietic stem cells. AB - Direct gene transfer to solid tissues or metastatic cancer cells requires vectors capable of in vivo transduction to specific cells. The predominant retroviral vectors of murine origin are inactivated by human complement, which precludes their use in vivo. Such inactivation does not take place with vectors based on human retroviruses. Murine retroviral vectors are also limited to proliferating cells, which human retroviruses are not. In this study we examined whether or not a vector using components from the human retroviruses HIV-1 and HTLV-1 could infect small-cell lung cancer cells and resting CD34+ hematopoietic stem cells. While HIV-1 itself was unable to infect cells lacking the CD4-membrane molecule, chimeric viral particles (pseudotype virus) with HIV-1 genome and HTLV-1 envelope components were able to infect both CD4-containing lymphocytic cells, CD4 negative tumour cells and hematopoietic stem cells. After infection with the pseudotype vector, the RNA genome was reverse transcribed and integrated. Transduction efficiency and gene expression under the HIV-1 LTR promoter in both tumour and stem cells were found to be of a similar or greater magnitude than in lymphocytic cells. These results suggest that gene transfer targeting proliferating as well as resting cells in vivo may be realized using components from human retroviruses. PMID- 9350218 TI - Ki-67 expression in relation to clinicopathological variables and prognosis in colorectal adenocarcinomas. AB - Ki-67 is a protein associated with cell proliferation which is expressed in all phases of the cell cycle except Go. In the present study, Ki-67 expression in 255 human colorectal adenocarcinomas was examined using immunohistochemistry with the monoclonal antibody MIB-1. One hundred and fifty-seven (62%) cases had more than 50% positive tumour cells and 98 (38%) cases less than 50%. The tumours showed a wide range of Ki-67 expression, from 13% to 90%, which indicated a variation in proliferative activity. There was no significant relationship between Ki-67 expression and sex, age, tumour location, Dukes' stage, growth pattern, differentiation, DNA content, S-phase fraction or survival (p > 0.05). In conclusion, the proliferative activity as measured by Ki-67 antibody was not related to clinicopathology and prognosis in colorectal cancer. PMID- 9350219 TI - The FEMA GRAS assessment of furfural used as a flavour ingredient. Flavor and Extract Manufacturers' Association. AB - The Expert Panel of the Flavor and Extract Manufacturers' Association (FEMA) has assessed the safety of furfural for its continued use as a flavour ingredient. The safety assessment takes into account the current scientific information on exposure, metabolism, pharmacokinetics, toxicology, carcinogenicity and genotoxicity. Furfural was reaffirmed as GRAS (GRASr) as a flavour ingredient under conditions of intended use based on: (1) its mode of metabolic detoxication in humans; (2) its low level of flavour use compared with higher intake levels as a naturally occurring component of food; (3) the safety factor calculated from results of subchronic and chronic studies, (4) the lack of reactivity with DNA; and (5) the conclusion that the only statistically significant finding in the 2 year NTP bioassays, an increased incidence of hepatocellular adenomas and carcinomas in the high-dose group of male mice, was secondary to pronounced hepatotoxicity. Taken together, these data do not indicate any risk to human health under conditions of use as a flavour ingredient. This evidence of safety is supported by the occurrence of furfural as a natural component of traditional foods, at concentrations in the diet resulting in a 'natural intake' that is at least 100 times higher than the intake of furfural from use as a flavour ingredient. PMID- 9350220 TI - Hepatic and associated response of rats to pregnancy, lactation and simultaneous treatment with butylated hydroxytoluene. AB - This paper describes changes in the livers of rats fed diets containing butylated hydroxytoluene (BHT) over two generations in two separate studies. BHT did not produce tumours when tested for carcinogenicity in several studies by the conventional way. However, when BHT was given to rats in a two-generation carcinogenicity study, a high incidence of hepatic tumours was found in males but not in female rats of the F1 generation. A sequential study has been carried out to gain an insight into this unexpected finding, paying particular attention to the perinatal period. In the dose-ranging study designed to assess the tolerance of rats to BHT, groups of male and female rats (F0 generation) were fed diets calculated to deliver 0, 500, 750 and 1000 mg/kg body weight/day. Following a loading period of 5 wk the rats were mated. The BHT content of the diet was not adjusted during pregnancy and lactation. Owing to the normal increase in food consumption during lactation, intakes peaked at double the nominal value by 21 days after the birth of pups. At this time the pups (F1) were weaned onto control diet and maintained on it for 4 wk. At birth, the body weights of pups from the BHT-treated dams were comparable to those of the controls but at weaning the body weights of the pups from all three dose levels were less than those of the controls. At the termination of the experiment (4 wk after weaning), the pups from BHT-treated dams still weighed less than those from untreated controls. In the main experiment the F0 generation were fed 0, 25, 100 and 500 mg/kg body weight/day. Their offspring (F1 generation) were weaned on diets containing the same amount of BHT as the respective parents, apart from the group given the highest dose level (500 mg/kg body weight/day). This dose level was reduced to 250 mg/kg body weight/day at weaning in order to conform with previously published findings. The pups from the dams given the highest dose level were maintained on a dietary concentration of 250 mg/kg body weight/day for the entire study. A group of age-matched non-pregnant females was also studied and the results obtained compared with those from pregnant dams. Pups from all groups were examined at day 20 of gestation, at weaning (21 days after birth), and at 4 and 22 wk post-weaning. There were no effects on fertility and no increase in foetal abnormalities at any dose of BHT. Dams receiving BHT at a nominal dose of 500 mg/kg body weight/day showed liver enlargement accompained by induction of pentoxyresorufin O-depentylase and glutathione S-transferase, and proliferation of the endoplasmic reticulum. Pups from these dams were of the same weight at birth as controls but lost weight during the lactation period. This deficit was not recovered by the time the experiment was terminated. Hence, in two independent studies, the only significant finding in rats treated with BHT in utero and during lactation was that the weight gain of pups during lactation was less than expected when dams received at least 500 mg BHT/kg body weight/day. The body weight of pups did not return to normal following a return to a control diet for 4 wk. It is postulated that the retardation in weight gain of the pups could be due to inadequate milk production. PMID- 9350222 TI - Identification of 2,5-dimethyl-4-hydroxy-3[2H]-furanone beta-D-glucuronide as the major metabolite of a strawberry flavour constituent in humans. AB - 2,5-Dimethyl-4-hydroxy-3[2H]furanone (Furaneol, DMHF) [3658-77-3], an important flavour constituent of strawberry fruit, was administered to four male and two female volunteers using fresh strawberries as a natural DMHF source. The amount excreted was determined by measuring urinary levels of DMHF and DMHF glucuronide. DMHF glucuronide was synthesized and the structure elucidated by mens of 1H, 13C and two dimensional nuclear magnetic resonance, as well as mass spectral data. Identification and quantification of DMHF glucuronide in human urine were achieved after solid phase extraction on XAD-2 using reverse-phase reverse-phase HPLC with either on-line UV/VIS or electrospray tandem mass spectrometry detection. Male and female volunteers excreted 59-69% and 81-94%, respectively, of the DMHF dose (total of free and glycosidically bound DMHF in strawberries) as DMHF glucuronide in urine within 24 hr. The amount of DMHF excretion was independent of the dose size and the ratio of free to glycosidically bound forms of DMHF in strawberry fruit. DMHF, DMHF glucoside and its 6'-O-malonyl derivative, naturally occurring in strawberries, were not detected in human urine. PMID- 9350221 TI - Effect of evening primrose oil on gastric ulceration and secretion induced by various ulcerogenic and necrotizing agents in rats. AB - The evening primrose oil (EPO) commercially known as Callanish evening primrose oil (omega-6 polyunsaturated fatty acid) is linoleic acid (LA) and gamma linolenic acid (GLA)-enriched oil obtained from the seeds of Oenothera biennis L. (Fam. Onagraceae). EPO was investigated for its ability to protect the gastric mucosa against injuries caused by pylorus ligation, non-steroidal anti inflammatory drugs (NSAIDs; aspirin, indomethacin and phenylbutazone), hypothermic restraint stress and necrotizing agents [0.6 M HCl, 0.2 M NaOH, 25% NaCl or 80% (v/v) aqueous ethanol]. It was administered by gastric intubation at doses of 5 and 10 ml/kg body weight to rats fed standard chow diet. An additional group of animals was given the same amount of corn oil in each experimental model studied. The results showed that EPO at the doses of 5 and 10 ml/kg body weight provided significant protection in various experimental models used. It produced a significant inhibition of gastric mucosal damage induced by pylorus ligation, NSAIDs, or hypothermic restraint ulcers. EPO also had a marked cytoprotective effective effect against all necrotizing agents used in this study. The results suggest that EPO rich in LA and GLA possesses both antisecretory and anti ulcerogenic effects. PMID- 9350223 TI - Lack of promotion of urinary bladder carcinogenesis by sodium bicarbonate and/or L-ascorbic acid in male ODS/Shi-od/od rats synthesizing alpha 2 mu-globulin but not L-ascorbic acid. AB - The study was designed to investigate whether sodium bicarbonate (NaHCO3) and/or L-ascorbic acid (AsA) promote urinary bladder carcinogenesis in male ODS/Shi od/od (ODS) rats, which, unlike male F344 rats, are resistant to sodium L ascorbate (Na-AsA)-promoting effects. Whereas F344 rats can synthesize AsA and alpha 2 mu-globulin (A2 mu-G), only A2 mu-G in produced in ODS rats. The two strains were given 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in their drinking water for 2 wk and then were fed basal CA-1 diet supplemented with 3% NaHCO3 plus 5% AsA (NaHCO3 + AsA), 3% NaHCO3, 5% AsA, or no chemicals for 32 wk. ODS rats given BBN-NaHCO3 or BBN-(NaHCO3 + AsA) had only a few small carcinomas in the urinary bladder, like those receiving BBN alone or BBN-AsA. In contrast, F344 rats administered BBN-NaHCO3 or BBN-(NaHCO3 + AsA) had many more, larger, carcinoma than animals of the same strain given BBN alone or BBN-AsA. AsA alone did not have any effect in either strain. Administration of NaHCO3 alone or NaHCO3 + AsA was associated with significant elevation of urinary pH and Na+ concentration to the same extent in both strains but, again, AsA alone was without effect. NaHCO3 + AsA and AsA alone increased the urinary concentration of total ascorbic acid in both strains but the observed levels wer lower in ODS rats. The results indicate that ODS rats are resistant to the modifying effects of NaHCO3 and/or AsA on two-stage urinary bladder carcinogenesis, and thus that the susceptibility to the promotional activity of sodium-salt-type compounds may be regulated by factors other than A2 mu-G-synthesizing ability and urinary levels of pH, Na+ and total ascorbic acid. PMID- 9350224 TI - Phenobarbital and 3-methylcholanthrene treatment alters phase I and II enzymes and the sensitivity of the rat colon to the carcinogenic activity of azoxymethane. AB - It has been hypothesized that cancer risk may be influenced by phase I and II drug-metabolizing enzyme systems. This study attempted to determine the relationship between colon phase I and II enzyme activity and the subsequent induction of aberrant crypt foci (ACF), preneoplastic lesions by azoxymethane (AOM), a colon-specific carcinogen. Phenobarbital (PB) and 3-methylcholanthrene (MC) treatment (prototype hepatic inducers of phase I and II enzymes) provided the framework to study the induction of phase I and II enzymes in the rat colonic mucosa. Following induction for five consecutive days, the animals were given a single injection of AOM. Phase I and II enzymes were determined fluorometrically and spectrophotometrically and ACF were identified microscopically. Phase I and II xenobiotic metabolizing enzymes were induced in the rat colonic mucosa by prototype hepatic inducers. A lower number of ACF and crypt multiplicity was observed in animals induced with MC than in those in the non-induced and PB groups. Altered levels of phase I and II enzymes in the colon during preinitiation stages were associated with modulation in the growth of ACF, putative preneoplastic lesions. PMID- 9350225 TI - Liver tumour-promoting effects of oxfendazole in rats. AB - To examine whether oxfendazole has tumour-promoting activity, a total of 100 male Fisher 344 rats were initiated with a single ip injection of 100 mg/kg of diethylnitrosamine (DEN) or given saline vehicle alone and starting 1 wk later given diet containing 500, 250, 100, 10 or 0 ppm of oxfendazole for 8 wk. Sub groups of five rats each from the DEN plus 250 and 0 ppm groups were killed after wk 1 of oxfendazole treatment and the remaining animals at wk 8. At the termination relative liver weights were significantly increased in the DEN initiated and non-initiated groups treated with 250 ppm and 100 ppm or more, respectively, compared with the corresponding controls values. Light microscopical examination showed centrilobular hepatocellular hypertrophy in all animals receiving 100 ppm or more. Electron microscopy also revealed marked increases in smooth endoplasmic reticulum in hepatocytes of the DEN plus 500 ppm group. Furthermore, induction of cytochrome P-450 (CYP) 1A1/2, 2B1/2 or 4A1 was observed in the DEN plus 100 ppm group, that of CYP 1A1/2 being most marked. A similar change in CYP 1A1/2 was seen in the DEN plus 10 ppm group. The numbers and areas of connexin 32 (Cx32)-positive spots per hepatocyte were also significantly decreased in a dose-dependent manner. Similar changes in liver weights, P-450 isozymes and Cx32 immunohistochemistry were already evident in the DEN plus 250 ppm group at wk 1. The number of placental form glutathione S transferase positive single cells was significantly increased in the DEN initiated groups treated with 250 ppm or more. The results therefore strongly suggest that oxfendazole exerts liver tumour promotion potential. PMID- 9350226 TI - Oxidative degradation and detoxification of mycotoxins using a novel source of ozone. AB - Practical methods to degrade mycotoxins using ozone gas (O3) have been limited due to low O3 production capabilities of conventional systems and their associated costs. Recent advances in electrochemistry (i.e. proton-exchange membrane and electrolysis technologies) have made available a novel and continuous source of O3 gas up to 20% by weight. It is possible that the rapid delivery of high concentrations of O3 will result in mycotoxin degradation in contaminated grains--with minimal destruction of nutrients. The major objectives of this study were to investigate the degradation and detoxification of common mycotoxins in the presence of high concentrations of O3. In this study, aqueous equimolar (32 microM) solutions of aflatoxins B1 (AfB1), B2 (AfB2), G1 (AfG1), G2 (AfG2), cyclopiazonic acid (CPA), fumonisin B1 (FB1), ochratoxin A (OA), patulin, secalonic acid D (SAD) and zearalenone (ZEN) were treated with 2, 10 and/or 20 weight% O3 over a period of 5.0 min and analysed by HPLC. Results indicated that AfB1 and AfG1 were rapidly degraded using 2% O3, while AfB2 and AfG2 were more resistant to oxidation and required higher levels of O3 (20%) for rapid degradation. In other studies, patulin, CPA, OA, SAD and ZEN were degraded at 15 sec, with no by-products detectable by HPLC. Additionally, the toxicity of these compounds (measured by a mycotoxin-sensitive bioassay) was significantly decreased following treatment with O3 for 15 sec. In another study, FB1 (following reaction with O3) was rapidly degraded at 15 sec, with the formation of new products. One of these appeared to be a 3-keto derivative of FB1. Importantly, degradation of FB1 did not correlate with detoxification, since FB1 solutions treated with O3 were still positive in two bioassay systems. PMID- 9350227 TI - Measurement of styrene oxide in polystyrenes, estimation of migration to foods, and reaction kinetics and products in food simulants. AB - The concentration of styrene-7,8-oxide has been measured in nine base resins and 16 samples of polystyrene articles intended for food contact. The epoxide was not detected in the resins (limit of detection 0.5 mg/kg) but was found in 11 of the 16 packaging samples at up to 2.9 mg/kg. Assuming that the propensity of styrene oxide to migrate is the same as styrene monomer, and using existing survey data for styrene monomer in packaging and foods, the migration levels expected for styrene oxide were calculated. Estimates were from 0.002 to 0.15 microgram/kg styrene oxide in foods. The stability of styrene oxide in the four standard EU food simulants was studied at 40, 100, 150 and 175 degrees C, to establish the transformation products to be expected following migration testing. The half-life at 40 degrees C in distilled water, 15% aqueous ethanol, 3% aqueous acetic acid and olive oil was 15, 23, < 1, > 2000 hr, respectively. The principal product was the diol from hydrolysis of the epoxide group. Ring opening in aqueous ethanol simulant gave the diol and also the glycol monoethyl ether. It is concluded that this instability of styrene oxide will reduce concentrations in foods, from an already low migration level to even lower levels with the formation of hydrolysis products that are less toxic than the parent epoxide. PMID- 9350228 TI - Brick tea consumption as the cause of dental fluorosis among children from Mongol, Kazak and Yugu populations in China. AB - Dietary fluoride intake and the prevalence of dental fluorosis were investigated in children from three population groups (mongol, Kazak and Yugu) in Gansu Province, China. The concentration of fluoride in drinking water ranged from 0.11 to 0.32 mg/litre; there was no other fluoride pollution. There was a high prevalence of dental fluorosis-52, 84 and 76% among the Mongol, Kazak and Yugu children, respectively). Dental fluorosis was particularly severe among the Kazak population (severity index: 2.00, 3.05 and 2.57 among the three populations, respectively). Each of the population groups had a long tradition of drinking milk tea made from brick tea water. This milk tea was found to contain high concentrations of fluoride (2.58-3.69 mg/litre). The daily fluorine consumption was 1.36-2.42-times the US RDA of 2.5 mg for children. Regression analysis showed that fluorosis was significantly correlated with the consumption of milk tea made from brick tea water, but not with any other dietary component (including milk). PMID- 9350229 TI - Metanil yellow: a bifunctional inducer of hepatic phase I and phase II xenobiotic metabolizing enzymes. AB - Metanil yellow, a non-permitted food colour, has been found in various foodstuffs. The induction potential of metanil yellow on hepatic microsomal cytochrome P-450 (P-450)-dependent monooxygenases and cytosolic detoxification enzymes, namely, glutathione S-transferase (GST) and quinone reductase (QR), was investigated. Oral administration of metanil yellow (430 mg/kg body weight) to four animals for seven days caused significant induction of hepatic P-450 (48%) and its dependent aryl hydrocarbon hydroxylase (100%) activity and cytosolic GST (136%) and QR (92%) activities. Parenteral administration of metanil yellow (80 mg/kg body weight) to another set of four animals for 3 days resulted in higher induction of ethoxyresorufin-O-deethylase (228%) as compared to other monooxygenases (64-92%), while GST and QR were also found to be induced (59-95%). Spectra of metanil yellow-induced microsomes showed an increase in P-450 with a shift of 2.2 nm in the soret region. The results suggest that metanil yellow acts as a bifunctional inducer of specific isozymes of P-450 and cytosolic enzymes and thus may involve the cytosolic aryl hydrocarbon (Ah) receptor for this type of induction. PMID- 9350230 TI - Gold: an allergen of growing significance. AB - Gold moved into the limelight of medical literature thanks to the anti inflammatory activity and effectiveness of gold compounds in the treatment of rheumatoid arthritis, but more recently also because of the growing incidence of hypersensitivity induced by it which is expressed in cutaneous and mucosal reactions. This review discusses dermatotoxicity associated with gold. In some countries gold has moved into second place as allergen, following nickel. Such recognition is mainly due to improved diagnostic methods and to its inclusion in routine dermal patch testing. Some unconventional manifestations of hypersensitivity are associated with use patterns which involve intimate contact with the metal as a component of jewelry. In-depth analysis of the growing number of cases of allergy has revealed various immunological idiosyncrasies as being characteristic of this metal. These include late reactions to challenge, extraordinary persistence of clinical effects, formation of intracutaneous nodules and immunogenic granuloma unresponsive to conventional steroid therapy, the occurrence of eczema at sites distant from the site of contact, and flare-ups of eczema upon systemic provocation with allergen which are characteristic of drug induced allergy. These manifestations demand investigations at the molecular level of the unusual mechanisms of action involved. PMID- 9350231 TI - The classification of skin irritants by human patch test. AB - The human 4 hour patch test provides an opportunity to identify substances with significant skin irritation potential without recourse to the use of animals. The protocol is designed to avoid the production of more than mild irritant reactions and meets the highest ethical standards. This paper provides the background to the development of the method and comments on its performance in the light of recent intra- and inter-laboratory investigations. In particular, the value of the method in providing 'gold standard' data for the identification of those substances (or preparations) which should, or should not, be classified as irritant to skin in European legislation is discussed. On the basis of the published data and supplementary investigations, recommendations are made on both the conduct and interpretation of the human 4 hour patch test. Finally, the lack of any necessity for formal validation of this assay is addressed. PMID- 9350232 TI - [Implementation of new Japanese GCP and the quality of clinical trials--from the standpoint of the pharmaceutical industry]. AB - As a part of the amendment of the pharmaceutical law and regulations in Japan, the GCP has been revised and enacted into the law and regulations. The new GCP is based on ICH-GCP and aims at upholding the quality of the Japanese clinical trials to an internationally acceptable level. The roles of the people involved in clinical trials have been renewed. The sponsor is responsible for management of clinical trials. At the same time, the procedures for quality assurance and quality control have extensively been brought into the new GCP. Not only monitoring but also audit of clinical trials are now the obligations of the sponsor. In this article are described major responsibilities newly assigned to the sponsor, medical institutions and investigators as well as the matters to be tackled. Discussion was also made extending the theme to matters other than GCP for upholding the quality of clinical trials in Japan to the internationally acceptable level. It is evident that we will confront many difficulties to solve these matters, but the implementation of the new GCP should give us a good chance to improve the quality of clinical studies performed in Japan. PMID- 9350233 TI - [Current status of low-dose CDDP. 5-FU therapy for solid malignant tumors- nationwide questionnaire survey]. AB - A nation-wide questionnaire survey was undertaken concerning low-dose anticancer therapy of CDDP plus 5-FU, which involves (5-10 mg CDDP/body/day + 300-500 mg/body/day) for 4-6 weeks. Out of 1,525 cases from 130 institutions, 847 cases with evaluable lesions were collected from 79 institutions. The response rate was 56.4% in esophageal cancer, 34.3% in gastric cancer, 35.3% in colorectal cancer, 47.2% in liver cancer and 35.9% in lung cancer, respectively. Adverse effects were found to be fewer and compliance was much better than the conventional therapy. Such figures suggest that the present regimen may be more effective than any so far. Problems for medical administration such as unlicensed CDDP for colorectal cancer were pointed out, which hinder the forthcoming third phase study. PMID- 9350234 TI - [Head and neck cancer]. AB - Phase II study of nedaplatin (NDP), a new derivative of cisplatin, was completed in 1990, so this agent is now commercially available. NDP is very effective for head and neck cancer. Out of the 90 evaluable patients, CR was achieved in 11 patients and PR in 27 with a response rate of 42%. A new combination chemotherapy containing NDP, especially NDP + 5-FU, was clinically tried. Furthermore concurrent NDP and radiotherapy will be tried in the near future. Phase II study of S-1 (tegafur + CDHP + Oxo) and taxotere (TXT), however, is ongoing. The results obtained so far are almost satisfactory. The aouthor also adopted several new agents which were presented at the ASCO meeting (1993-1997): taxol (TXL), taxotere (TXT), topotecan, amonafide, vinorelbine and thymitaq. Response rates of these agents were as follows: TXL: 26-37%, TXT: 27-41%, topotecan: 0-27%, vinorelbine: 6.7-12.5%, thymitaq: 18.2% and amonafide: 3.6%. So TXL and TXT are very effective for head and neck cancer. In terms of combination chemotherapy, response rates are 33-71% in TXL + CDDP, 23-62% in TXL + CBDCA, 78% in TXT + CDDP and 75% in TXT + CDDP + 5-FU. Concurrent radiotherapy and chemotherapy including new agents are interesting and important issues. Two kinds of protocol were adopted, 5-FU + HU + TXL + RT and TXL + CBDCA + RT. Both protocols are responsive to squamous cell carcinoma of the head and neck. But severe local toxicity (stomatitis) and bone marrow suppression pose problems. PMID- 9350235 TI - [New drugs in metastatic breast cancer--1997]. AB - The introduction of new drugs may offer significant hope for improvement in the treatment of breast cancer. They include new cytotoxic agents such as Taxanes, Topoisomerase inhibitor, MDR modulator, new hormone such as 3rd generation nonsteroidal aromatase inhibitor, pure antiestrogen for patients with refractory metastatic breast cancer and new bisphosphonates for those who have metastases to bone. Therefore, some reports evaluating such new drugs were reviewed. Japanese studies revealed response rate of Taxanes in the treatment of metastatic breast cancer as follows; Six out of 22 (27%) with 1 CR for 24 hours infusion, 21 out of 52 (34%) with 2 CR for 3 hours infusion of paclitaxel respectively, 21 out of 31 (41%) with 2 CR for 1 hour infusion, and 32 out of 72 (44%) with 5 CR for 1 hour infusion of Docetaxel, In addition, the MTD of the combination was reached at dose level with 60 mg/sqm of Docetaxel and 400 mg/sqm of cyclophosphamide. As regards hormone therapy, thirty four out of 176 (19%) with 5 CR were observed in phase II study of Fadrozol but results from the phase II study of 3rd generation aromatase inhibitor is not yet accomplished in Japan. However, any european reports did not show a difference in response rate in patients with tamoxifen refractory breast cancer between newer aromatase inhibitor and megasterol, and a good choice of hormones in the treatment of metastatic breast cancer appears to be difficult. PMID- 9350236 TI - [Lung cancer]. AB - Recently, a number of novel chemotherapeutic agents under development show consider able promise in the treatment of lung cancer. Optimally, these new agents should exhibit a unique mechanism of action, a favorable toxicity profile in comparison to standard cisplatin-based chemotherapy, significant single agent activity (> 20% response rate, or > 40% 1-year survival rate) and synergistic antitumor effects with cisplatin and/or radiation. This review will describe the current development status of these new agents, including paclitaxel, docetaxel, vinorelbine, CPT-11, topotecan and gemcitabine. PMID- 9350237 TI - [New anti-cancer drugs for gastrointestinal cancers]. AB - Promising new drugs for gastrointestinal cancers in clinical phase II studies in Japan were reviewed. Treatment with l-Leucovorin (l-LV, combined with 5-FU), S-1, a methionine-free intravenous amino acid solution (AO-90), BOF-A2, and interferon (combined with 5-FU), is based on biochemical modulation-related 5-FU. With combination of l-LV and 5-FU, response rates of over 30% were reported in clinical rate phase II studies for gastric and colorectal cancers respectively. S 1, a new oral tegaful pulse modulator, showed a response rate of 53.6% (in eligible cases) in early phase II studies for gastric cancer. Except for other than biochemical modulation drugs, 254-S (new cisplatin derivatives), CPT-11 (camptothecin derivatives) and docetaxel are also promising for gastrointestinal cancers. These drugs showed activity against advanced gastrointestinal cancers, suggesting that further clinical studies of these drugs combined with other active chemotherapy agents are warranted. PMID- 9350238 TI - [Promising new drugs for gynecological cancer]. AB - Five promising new drugs for gynecological cancer were reviewed. Taxans (Paclitaxel: Taxol and Docetaxel: Taxotere) diterpenoid plant products enhance the polymerization of tublin. Taxol showed significant activity for platinum refractory ovarian cancer in a phase 1 clinical trial in the United States. The combination with cisplatin (CDDP) showed superior results to CDDP plus Cyclophosphamide and has been recognized as a new standard in adjuvant chemotherapy for advanced ovarian cancer. The major toxicities are myelosuppression, alopecia, and hypersensitivity reactions (HSRs). HSRs were overcome by pretreatment with anti-histamines and over 24 hours administration. It was also reported that Taxol was administered safely by over 3 hours infusion with reduced myelotoxicity, but the incidence of HSRs may be increased. Clinical trials of intraperitoneal administration and combination with Carboplatin (CBDCA) are ongoing. Taxotere, an analog of Taxol, is also effective as Taxol with a low incidence of HSRs. Topoisomerase inhibitors (Irinotecan hydrochloride: CPT-11 and Topotecan) have promising antitumor activity for ovarian and cervical cancer. CPT 11 is a semisynthetic camptothesin analog developed in Japan. It was also effective for platinum-resistant ovarian cancer, such as mucinous and clear cell carcinoma. An adverse effect was observed in the combination of CPT-11 and CDDP. The phase 1 clinical trial showed a 40% response rate against recurrent ovarian cancer. CPT-11 50-60 mg/m2 (day 1,8,15) and CDDP 50-60 mg/m2 (day 1) are a recommended schedule. The major toxicities are neutropenia and diarrhea. Thrombocytopenia is not severe and diarrhea is also controllable. Topotecan is also a promising topoisomerase inhibitor and reported superior result to Taxol for platinum refractory ovarian cancer. A phase II trial is ongoing for ovarian and cervical cancer in Japan. Nedaplatin, a new analog of cisplatin, has similar activity especially for cervical cancer with less myelotoxicity and nephrotoxicity. PMID- 9350239 TI - [Development of new drugs for urological cancers]. AB - Urological cancers have a variety of biological characteristics. Thus, anti cancer agents in this field are often developed focusing on the biological characteristics. For example, LH-RH agonist and anti-androgens have been developed for androgen-sensitive prostate cancer, interferons and IL-2 for renal cell cancer, and BCG for superficial bladder cancer. In this manuscript, new agents which are thought to be promising in the urological field, bropirimine for bladder cancer and liarozole for prostate cancer, are briefly introduced. PMID- 9350240 TI - [Expression of ATP binding cassette superfamily (multidrug resistance-1, multidrug resistance-associated protein, human canalicular multispecific organ anion transporter) mRNA in etoposide and m-AMSA resistant cell lines]. AB - The efficacy of all chemotherapeutic agents is limited by the occurrence of drug resistance. To further understand resistance to topoisomerase (topo) II inhibitors, 50 sublines were isolated as single clones from parental cells by exposure to etoposide or m-AMSA. Subsequently, a population of cells from each sublines was exposed to three-fold higher drug concentrations allowing 16 stable sublines to be established at higher extracellular drug concentration. Quantitative aspects of MRP and C-MOAT were studied by Northern blotting in 66 resistant cell lines. Increased MRP mRNA was observed in 48.5% of resistant cell lines (64.7% of etoposide resistant cells and 31.3% of m-AMSA resistant cell lines). Increased C-MOAT mRNA was also observed in 39.4% of resistant cell lines (41.2% in etoposide resistant cell lines and 37.5% in m-AMSA resistant cell lines). To characterize the function of C-MOAT, cellular accumulation assay for 3H-etoposide was performed in three resistant cell lines which overexpress C-MOAT but do not express MRP. Accumulation of etoposide was reduced in the cell lines. Our findings suggest that increased MRP and O-MOAT mRNA seems to be an important mechanism of resistance to topo II inhibitors. PMID- 9350241 TI - [Effect on 5'-deoxy-5-fluorouridine (5'-DFUR) of pyrimidine nucleoside phosphorylase (PyNPase), matrix metalloprotease and serum IAP values. Hokuriku Colorectal Cancer Chemotherapy Study Group]. AB - PyNPase activity, MMPs activity and serum IAP values were measured in tumor tissues from colorectal cancer patients who had been divided into two groups, one given preoperative 5'-DFUR and the controls. PyNPase activity of the preoperative administration group was approximately equivalent to that of the controls. In the control group, correlations were assessed between PyNPase activity and activities of MMP1 and MMP3. To assess the effect of 5'-DFUR on the activity of MMPs, we divided patients into two groups, a high and a low PyNPase activity group. Although there was no correlation with MMPs activity of the preoperative administration group and the control group in the low PyNPase activity group, the activities of MMP1 and MMP9 of the control group were significantly higher in the high PyNPase activity group. Moreover, the serum IAP value of the administration group was significantly lower than that of the control group. These results indicated that PyNPase activity was thus suggested to be somehow related to MMPs activity and serum IAP values. PMID- 9350242 TI - [Prospective controlled study on the usefulness of Carmofur as a postoperative adjuvant chemotherapy for colorectal cancer]. AB - A prospective controlled study, the 7th cooperative study of the Japanese Foundation for Multidisciplinary Treatment, was conducted to evaluate the usefulness of concomitant therapy with MMC + HCFU as a postoperative adjuvant therapy in patients with colorectal cancer who had undergone curative resection for a period of 2 years and 11 months from February, 1986. The Dukes B and C patients with colorectal cancer classified by macroscopic examination who had an intravenous MMC 6 mg/m2 on the day of operation and followed by oral HCFU for 12 months from 2 weeks after operation (Group X) were compared with patients who had operation only (Group Y). Some 978 patients with colon cancer and 713 patients with rectal cancer were enrolled in the study, 85 (5.0%) of whom were not eligible. The 5-year survival rate of Group X in colon cancer was 79.3% and that of Group Y was 76.4%: thus the survival rate of Group X was slightly better than that of Group Y, but no significant difference was found between the two groups. Subset analysis revealed that the survival rate of Group X in advanced cancer of stage III b + IV, according to the General Rules for Clinical and Pathological Studies on Cancer of the Colon, Rectum and Anus in Japan, was 62.4% and that of Group Y was 46.2%. Thus, the survival rate of Group X was significantly better than that of Group Y (logrank test: p = 0.035, generalized Wilcoxon test: p = 0.025). The disease-free survival rate was not significantly different between the groups with colon cancer and rectal cancer. The above results suggest that HCFU is useful for patients with a high risk of recurrence who had advanced colon cancer (stage III b + IV). However, additional prospective studies are required to verify them. PMID- 9350243 TI - [Thermoradiotherapy combined with daily administration of low dose cisplatin for locally advanced laryngeal carcinoma]. AB - We applied thermoradiotherapy combined with daily administration of low dose cisplatin (CDDP) to five patients with locally advanced laryngeal carcinoma. The total response rate (CR + PR) was 100% after irradiation of 30 or 40 Gy. One case showed CR, and the larynx could be preserved by adding a full dose of irradiation. CDDP administration was discontinued in two cases due to renal dysfunction or thrombopenia, but other cases tolerated the therapy without severe side effects. Total laryngectomy and bilateral neck dissection was carried out in four cases after irradiation of 30 or 40 Gy. Removed larynxes were prepared for light microscopic observations of the serial sections. One of the four specimens revealed a CR histologically. The combination of thermoradiotherapy with low dose CDDP showed a remarkable effect on reduction of tumor size and cytotoxicity of tumor cells. This might contribute to saving the larynx of patients with locally advanced laryngeal carcinoma. PMID- 9350244 TI - [Study on the most appropriate time for Romurtide administration (Nopia) in radiotherapy patients]. AB - The most appropriate time for administration was studied based on examination of the total number of leukocytes, differential white blood count, and the number of platelets in 30 cases in which 200 micrograms Romurtide (Nopia) was injected subcutaneously on the day of radiotherapy, 5 times a week, for 2 weeks, totally 10 times because leukopenia was caused during treatment with radiotherapy. It was recognized that the number of leukocytes, mainly neutrophils, increased from 1 week after starting administration of Romurtide to after the completion of administration (2 weeks after administration), except 1 week after completion of administration. The increasing effect in the number of platelets was not recognized, and it had decreased from at the completion of administration to 1 week after the completion of administration of Romurtide. The rate of completion of radiotherapy was 100% without any serious adverse events, but there were 4 cases in which fever was observed. Therefore, it is not very favorable to administer Romurtide immediately after radiation, taking account of the treatment period of radiotherapy. It is also considered that it would be necessary to start radiation after sufficient recovery of the number of leukocytes or increase in their number by using G-CSF preparation in cases in which the leukocyte count has fallen since starting radiation due to the influence of the preceding chemotherapy. PMID- 9350245 TI - [Histopathological effect of neoadjuvant chemohormonal therapy for prostatic cancer]. AB - To improve the therapeutic results in prostatic cancer, radical prostatectomy or total cystoprostatectomy were performed with chemohormonal therapy before operation. Radical prostatectomies were conducted in eight patients with localized prostatic cancer and total cystoprostatectomies in ten patients with severe cystic infiltration. The administration schedule of chemohormonal therapy was as follows: prior to operation, 30-60 mg/sqm/day of etoposide was administered for 7 days every 3 weeks, 250-500 mg/day of diethylbestrol diphosphate for 30 days, and 3.6 mg of LH-RH agonist was also administered. Sixteen of the subjects survived, and were socially rehabilitated (14 cases of NED, 1 case of NC and 1 case of PD) and 2 of the subjects died of cancer. Histopathological findings showed 9 cases of poorly differentiated adenocarcinoma, 4 cases of well differentiated adenocarcinoma and 5 cases of moderately differentiated adenocarcinoma. Histopathological effect of neoadjuvant chemohormonal therapy in surgical specimen showed that 2 of the subjects had grade 0a effect, grade 0b in 7 cases, grade 1 in 5 cases and grade 2 in 4 cases. PMID- 9350246 TI - [Evaluation of a new anti-cancer drug regimen against uterine cervical cancer in nude mice]. AB - The purpose of this study was to describe a new anti-cancer drug regimen for uterine cervical cancer. The cytotoxicities of some anti-cancer drugs regimens against the human uterine cervical cancer xenografted into nude mice have been studied. The activities of CDDP, CPT-11, TXL, CDDP + BLM, CDDP + MMC, CPT-11 + BLM, CPT-11 + CDDP, CDDP + 5-FU, CPT-11 + MMC, CPT-11 + TXL and CDDP + TXL for squamous cell carcinoma (TCR, TCK, TCG), and CDDP, MMC, TXL, CDDP + TXL, CDDP + MMC and MMC + TXL for adenocarcinoma (TCO, TCM, TCY), were evaluated comparing with a control group using saline. Five mice were used for each groups. When the xenografted tumor reached 6 mm in diameter, 1/5 LD50 of these drugs were administered into the peritoneal cavity of the mice once a week for three weeks. The effective regimens were CDDP + MMC, CDDP + BLM, CDDP + CPT-11 and CPT-11 + MMC for squamous cell carcinoma of the uterine cervix. CDDP + MMC, CDDP + TXL, MMC + TXL and CDDP were effective for endocervical adenocarcinoma. It was suggested that these new drug regimens should be used in clinical studies. PMID- 9350247 TI - [Efficacy of systemic chemotherapy in adenocarcinoma of the lung with pleuritis carcinomatosa]. AB - The efficacy of systemic chemotherapy (CDDP + IFO + 5-FU or CDDP + IFO + CPT-11) was evaluated in 41 patients with malignant pleural effusion secondary to adenocarcinoma of the lung. The overall response rate for measurable disease was 56.1%. The response for pleural effusion was evaluated according to the criteria of the Japan Lung Cancer Society. The overall response for pleural effusion was 53.7% (34.1% CR and 19.5% PR). The median survival time was 361 days. These results suggested that systemic chemotherapy is an effective treatment for pleuritis carcinomatosa. PMID- 9350248 TI - [Cardiotoxicity due to prolonged administration of THP (2"R-4'-O tetrahydropyranyl-adriamycin)]. AB - The effect of the anthracycline analogue, pirarubicin, on cardiac function was examined. One hundred and four patients with gynecologic malignancies were treated with 40-50 mg/body THP ADM every 3 to 4 weeks by iv bolus injection. Three out of 104 cases that had developed cardiac failure received more than 1,500 mg. One case receiving 1,340 mg developed cardiac failure and expired 3 years after completion of treatment for malignancy. From the above experience, it is concluded that the MTD of pirarubicin seems to be 1,500 mg/body or 1,100 mg/m2. PMID- 9350249 TI - [A case of local advanced cervical esophageal cancer treated with concurrent radiochemotherapy followed by surgery]. AB - A 61-year-old-male with local advanced inoperable cervical esophageal cancer (squamous cell carcinoma, T4N1M0 stage III) was treated by concurrent radiochemotherapy. A dose of 50.6 Gy/46 Fr/36 days and one course each of CDDP-5 FU and nedaplatin-5-FU were delivered safely. Radical surgery could be performed thereafter because of the good tumor response. Pathological CR was obtained in metastatic cervical lymph nodes and almost total necrosis in primary cancer. Concurrent radiochemotherapy produced a significant improvement in this case of advanced cervical esophageal cancer. PMID- 9350251 TI - [Cisplatin/5-fluorouracil intraperitoneal administration superior to intravenous administration for liver metastases of colonic carcinoma]. AB - A 28-year-old female underwent sigmoidectomy for sigmoid colon cancer with peritoneal seeding. One month later, a solitary metastasis was found in S3 of the liver. After CDDP/5-FU intravenous chemotherapy, another metastasis appeared in S7. Intravenous administration showed PD. But the metastatic tumors shrank and became inobservable by CT after the 1st round of CDDP/5-FU intraperitoneal chemotherapy, and S7 tumor could not be identified after the 2nd round. Many previous reports demonstrated the concentration of cytotoxic drug in intraperitoneal administration was much higher than in intravenous administration. Theoretically, intraperitoneal chemotherapy is superior to intravenous chemotherapy for the prevention and treatment of liver metastases. This case demonstrated this hypothesis was right. We think adjuvant intraperitoneal chemotherapy should be re-considered for the prevention of the liver metastases of gastrointestinal cancers. PMID- 9350250 TI - [A case of papillary adenocarcinoma of the stomach with liver metastases and carcinomatous peritonitis treated effectively by methotrexate/5-fluorouracil sequential chemotherapy]. AB - A 54-year-old male was admitted to receive treatment by anticancer drug for advanced gastric cancer with liver metastases and carcinomatous peritonitis. Upper gastrointestinal series showed a Borrmann type 4 gastric cancer occupying the body and antrum, and a pathological diagnosis of papillary adenocarcinoma was made by endoscopic biopsy. Two cycles of neoadjuvant chemotherapy consisting of 5 FU and low-dose CDDP (FP therapy) were given. No effect was observed, so that the next chemotherapy schedule of sequential MTX/5-FU therapy was performed. We adopted the intermediate (MTX: 100 mg/m2 and 5-FU 600 mg/m2 weekly) dose regimen. When six courses of this regimen were completed, the primary lesion of the stomach was decreased to 60% and the metastatic liver tumor to 72%, basel on criteria for the evaluation of clinical effects of solid cancer chemotherapy. However, pylorous stenosis remained, and we performed gastrojejunostomy and biopsy of the regional lymph node to determine the response to the chemotherapy. A pathological examination of lymph node revealed metastatic papillary adenocarcinoma with xanthogranulomatous change, and was judged as an effect showing grade 2. He was discharged and had the outpatient hospital treatment. He died of exacerbation of carcinomatous peritonitis 10 months after his initial admission. The response duration for sequential MTX/5-FU therapy was 4 months. This therapy was usually effective in patients with poorly differentiated adenocarcinoma of the stomach. Our result indicates that this therapy can be effective for not only poorly differentiated adenocarcinoma but also papillary adenocarcinoma of the stomach in an advanced stage. PMID- 9350252 TI - [Colon and rectum]. AB - The present TNM classification of the large bowel cancer which appeared in the "TNM Classification of malignant Tumours. Fourth Edition 2nd Revision 1992" was explained. The differences of T classification among large intestine and stomach and also depth of invasion classification of the Japanese Society for Cancer of the Colon and Rectum (JSCCR) was also explained. Using TNM Supplement 1993, detailed explanation was given for some delicate terminological problems such as pericolic nodes, perirectal nodes and lymph nodes along the course of the named vascular trunk. The new classification of cancer of the small intestine and a proposal for a classification of the gastrointestinal sarcoma were introduced. PMID- 9350253 TI - Drugs in salmonid aquaculture--a review. AB - In contrast to mammalian therapeutics, the use of pharmaceutical substances is rather limited in fish. It is basically restricted to anaesthetic agents and anti infective agents for parasitic and microbial diseases. Anaesthetic agents are used primarily in fish farm and laboratory settings to provide analgesia and immobilization of fish for minor procedures. The anti-infective agents are used for controlling diseases and the choice of drug depends on efficacy, ease of application, human safety, target animal safety including stress to the fish, environmental impact, regulatory approval, costs, and implications for marketing the fish. In this article, the major drugs used in salmonids in North America and Europe will be reviewed and some insight into future directions for drug development and use for the salmonid industry will be introduced. The mechanisms of action, pharmacokinetics, side effects, and uses of the drugs are emphasized. PMID- 9350255 TI - Pharmacokinetics of intravenous and intragastric cimetidine in horses. I. Effects of intravenous cimetidine on pharmacokinetics of intravenous phenylbutazone. AB - Cimetidine was administered intravenously and by the intragastric route to six mares at a dose of 4.0 mg/kg of body weight (bw). Specific and sensitive high performance liquid chromatographic methods for the determination of cimetidine in horse plasma and urine and cimetidine sulfoxide in urine are described. Plasma cimetidine concentration vs. time data were analysed by non-linear least squares regression analysis to determine pharmacokinetic parameter estimates. The median (range) plasma clearance (Cl) was 8.20 (4.96-10.2) mL/min.kg of body weight, that of the steady-state volume of distribution (Vdss) was 0.771 (0.521-1.15) L/kg bw, and that of the terminal elimination half-life (t1/2 beta) was 92.4 (70.6-125) minutes. The median (range) renal clearance of cimetidine was 4.08 (2.19-6.23) mL/min.kg bw or 55.4 (36.3-81.8)% of the corresponding plasma clearance. Cimetidine sulfoxide was excreted in urine and its urinary excretion through 8 h accounted for 12.0 (9.8-16.6)% of the plasma clearance of cimetidine. The median (range) extent of intragastric bioavailability was 14.4 (6.82-21.8)% and the maximum plasma concentration after intragastric administration was 0.31 (0.24 0.50) microgram/mL. Intravenous cimetidine had no effect on the disposition of intravenous phenylbutazone or its metabolites except that the maximum plasma concentration of gamma-hydroxyphenylbutazone was less after cimetidine treatment. PMID- 9350254 TI - Reversal of medetomidine-induced sedation in reindeer (Rangifer tarandus tarandus) with atipamezole increases the medetomidine concentration in plasma. AB - The pharmacokinetics of two potent alpha 2-adrenoceptor agents that can be used for immobilization (medetomidine) and reversal (atipamezole) of the sedation in mammals, were studied in three reindeer (Rangifer tarandus tarandus) in winter and again in summer. Medetomidine (60 micrograms/kg) was injected intravenously (i.v.), followed by atipamezole (300 micrograms/kg) intravenously 60 min later. Drug concentrations in plasma were measured by HPLC. The administration of atipamezole resulted in an immediate 2.5-3.5 fold increase in the medetomidine concentration in plasma. Clearance for medetomidine (median 19.3 mL/min.kg) was lower than clearance for atipamezole (median 31.0 mL/min.kg). The median elimination half-lives of medetomidine and atipamezole in plasma were 76.1 and 59.9 min, respectively. The animals became resedated 0.5-1 h after the reversal with atipamezole. Resedation may be explained by the longer elimination half-life of medetomidine compared to atipamezole. PMID- 9350257 TI - Pharmacokinetics of ceftiofur and metabolites after single intravenous and intramuscular administration and multiple intramuscular administrations of ceftiofur sodium to dairy goats. AB - Twelve (12) lactating dairy goats (46-71 kg body wt at study initiation) were divided into four treatment groups and dosed with ceftiofur sodium at 1.1 mg ceftiofur free acid equivalents (CFAE)/kg or 2.2 CFAE/kg using a complete two route (intravenous, i.v.; intramuscular, i.m.), two-period crossover design, with a 2-week washout between injections. After another 2-week washout period, the goats were dosed with ceftiofur sodium i.m. for 5 consecutive days at either 1.1 or 2.2 mg CFAE/kg. The goats from the 2.2 mg/kg multiple dose group were dried off and the i.v. kinetic study repeated. After all injections, blood samples were obtained serially for determination of combined serum concentrations of ceftiofur and metabolites. After intravenous doses of 1.1 and 2.2 mg/kg, the harmonic means of the terminal phase half-lives were 171.8 and 233 min, respectively, for lactating does. The harmonic mean of the terminal phase half-life after an i.v. dose of 2.2 mg/kg in non-lactating does was 254 min. The AUC0-infinity was significantly less and the clearance significantly greater during lactation. After i.m. doses of 1.1 and 2.2 mg/kg, the harmonic mean terminal phase half lives were 163 and 156 min, respectively. The i.m. bioavailability of ceftiofur sodium in goats was 100%, and the AUC0-infinity was dose-proportional from 1.1 2.2 mg CFAE/kg body weight. After five daily i.m. doses of ceftiofur sodium at either 1.1 or 2.2 mg CFAE, there was minimal accumulation of drug in serum as assessed by Cmax, and serum concentrations were dose-proportional after the multiple dosing regimen. PMID- 9350256 TI - Pharmacokinetics of an injectable sustained-release formulation of morphine for use in dogs. AB - This study investigated the pharmacokinetics of morphine sulphate in an injectable chitosan-based gel. Gels were made from a combination of N-O carboxymethylchitosan (NOCC) and chitosan and were easily injectable via a 22 gauge needle and appeared stable during long-term storage. Groups of six beagles were injected subcutaneously (s.c.) with 1.2 mg/kg morphine sulphate, either in sterile saline or in sterilized gels, and serial blood samples were withdrawn via a jugular catheter and later analysed for morphine concentrations using radioimmunoassay. Data were analysed according to non-compartmental pharmacokinetics. NOCC-based gels resulted in significantly lower serum morphine concentrations at 10 and 30 min following injection but significantly higher concentrations at all points from 120 to 480 min post-injection. Dogs receiving morphine gel exhibited equivalent or lesser variability in serum morphine concentrations than dogs receiving conventional morphine sulphate. Pharmacokinetic analysis revealed that morphine release from the gel matrix was significantly prolonged but fully bioavailable. There were no significant differences in either distribution (Vd) or terminal elimination (t 1/2). Dogs experienced no adverse effects other than those normally associated with morphine administration at the time of injection but all dogs receiving the gel presented with an undefined stiffness the next day that resolved spontaneously within 48 h. We conclude that carboxymethylchitosan-based gels hold considerable promise for the development of injectable sustained-release formulations of opioid analgesics. PMID- 9350258 TI - The withdrawal time estimation of veterinary drugs: a non-parametric approach. AB - The study of a veterinary drug for use in a food producing animal requires the estimation of the withdrawal time which is defined as the time when it can be assessed with a controlled risk that a given percentage (less than 100(1-alpha)%) of animals have a concentration below the Maximal Limit of Residue (MRL). A simple and understandable non-parametric method which allows the control of the risk is presented. This method does not require the assumptions needed for the correct use of the current method of estimation to be met. A simple example which illustrates the use of this method is presented. PMID- 9350259 TI - The withdrawal time estimation of veterinary drugs revisited. AB - The aim of this article is to review current issues concerning the statistical determination of a withdrawal time for a veterinary drug. Special attention has been paid to the Food and Drug Administration (FDA) approach which consists of using a regression method to estimate a 99th percentile of the population with a 95% confidence level. The different assumptions of the simple regression techniques are reviewed. It is shown, both on theoretical grounds and using Monte Carlo simulations, that the main assumptions for the correct use of linear regression do not hold when a tissue depletion curve is under investigation. Finally, it is suggested that methods other than the regression technique must be evaluated, especially a revised version of the so-called simpler approach, which consists of using a non-parametric strategy to estimate a withdrawal time. PMID- 9350260 TI - Effects of a selective and a nonselective muscarinic cholinergic antagonist on heart rate and intestinal motility in dogs. AB - The effects of methoctramine, a cardioselective muscarinic cholinergic antagonist, on heart rate and small intestinal motor activity were compared to those of the nonselective competitive muscarinic antagonist, atropine. Methoctramine or atropine, 6, 10, 30, 60 micrograms/kg, or sterile isotonic saline, was administered intravenously to six conscious dogs in cross-over studies. Methoctramine administration caused dose-dependent tachycardia without affecting intestinal motility, while atropine administration caused dose dependent tachycardia accompanied by significant reductions in small intestinal motility. Additionally, methoctramine did not inhibit intestinal smooth muscle contractile activity initiated by the muscarinic agonist bethanechol, while atropine inhibited bethanechol-induced contractile activity in a dose-dependent manner. Calculated, dosages of methoctramine and atropine required to produce a 50% increase in heart rate over baseline were 35.1 +/- 5.3 and 39.5 +/- 6.2 micrograms/kg, respectively. This dosage of atropine caused a 93 +/- 13.9% reduction in intestinal motility. These findings suggest that selective muscarinic antagonists may be useful drugs for those veterinary patients in which nonselective muscarinic antagonists have the potential to produce untoward effects on intestinal motility. PMID- 9350261 TI - Development and characterization of an equine behaviour chamber and the effects of amitraz and detomidine on spontaneous locomotor activity. AB - This report describes the development of a behaviour chamber and the validation of the chamber of measure locomotor activity of a horse. Locomotor activity was detected by four Mini-beam sensors and recorded on a data logger every 5 min for 22 h. Horses were more active during daytime than in the evening, which was at least partially related to human activity in their surroundings. To validate the ability of the chambers to detect changes in activity, fentanyl citrate and xylazine HCl, agents well-characterized as a stimulant and a depressant, respectively, were administered to five horses. Fentanyl citrate (0.016 mg/kg) significantly increased locomotor activity which persisted for 30 min. Xylazine HCl (1 mg/kg) significantly reduced locomotor activity for 90 min. Amitraz produced a dose-dependent decrease in locomotor activity, lasting 75 min for the 0.05 mg/kg dose, 120 min for the 0.10 mg/kg dose, and 180 min for the 0.15 mg/kg dose. In a separate experiment, yohimbine administration immediately reversed the sedative effect of amitraz. This suggests there is a similarity in the mode of action of amitraz, xylazine and detomidine, as yohimbine acts primarily by blocking central alpha 2 -adrenoceptors that are stimulated by agents like xylazine. There was also a significant decrease in locomotor activity following injection of detomidine (0.02, 0.04 and 0.08 mg/kg) for 1.5, 3.5 and 5.0 h, respectively. The locomotor chamber is a useful, sensitive and highly reproducible tool for measuring spontaneous locomotor activity in the horse, which allows investigators to determine an agent's average time of onset, duration and intensity of effect on movement. PMID- 9350262 TI - Concentration of enrofloxacin in equine tissues after long-term oral administration. PMID- 9350263 TI - Pharmacokinetics of pefloxacin in sheep. PMID- 9350264 TI - Comparison of halothane minimum alveolar concentration and minimum effective concentration in ponies. PMID- 9350265 TI - The effect of praziquantel on the hepatic activities of some drug-metabolizing enzymes in rabbits experimentally infected with Schistosoma bovis. PMID- 9350266 TI - Recent studies on the effects of tiamulin and monensin on hepatic cytochrome P450 activities in chickens and turkeys. PMID- 9350280 TI - Viruses and autoimmune diseases. PMID- 9350281 TI - Induction of HIV-1 specific mucosal immune responses by DNA vaccination. AB - DNA vaccination has been shown to induce immunity against several different pathogens including HIV-1. The authors demonstrate here that administration of DNA vaccines via the intranasal route is sufficient to induce immune responses both at distal mucosal sites and systemically. Since transmission of HIV-1 occurs largely across mucosal surfaces, the intranasal route provides a further means of application for DNA immunization. PMID- 9350282 TI - Serum amyloid P component binds to influenza A virus haemagglutinin and inhibits the virus infection in vitro. AB - Serum amyloid P component (SAP) is a member of the phylogenetically conserved and structurally related group of proteins called pentraxins. SAP exhibits multispecific calcium-dependent binding to oligosaccharides with terminal N acetyl-galactosamine, mannose and glucuronic acid. The authors report that SAP can bind to influenza A virus and inhibit agglutination of erythrocytes mediated by the virus subtypes H1N1, H2N2 and H3N2. SAP also inhibits the production of haemagglutinin (HA) an the cytopathogenic effect of influenza A virus in MDCK cells. The binding of SAP to the virus requires physiological calcium concentrations and is blocked by specific SAP antibodies. Denaturated and renaturated SAP retained inhibition of HA. Electron microscopy shows Ca(2+) dependent binding of SAP to spikes on the viral envelope and immunoblotting indicates that SAP binds to a 50-55 kDa peptide corresponding to the mass of the HA1 peptide. Of several monosaccharides tested only D-mannose interfered with SAP's inhibition of both HA and infectivity. The glycosaminoglycans heparan sulfate and heparin, which bind SAP, reduced SAPs binding to the virus. The results indicate that the inhibition by SAP is due to steric effects when SAP binds to terminal mannose on oligosaccharides localized close to the sialic acid binding site of the HA trimer. PMID- 9350283 TI - Interferon-gamma enhancement of E-selectin expression on endothelial cells is inhibited by monensin. AB - The expression of E-selectin reaches a maximum 4-6 h after stimulation of human umbilical vein endothelial cells (HUVEC) in vitro with tumour necrosis factor alpha (TNF-alpha) and then declines to basal level within 24 h. If interferon gamma (IFN-gamma) is added to the cell culture medium together with TNF-alpha the surface expression of E-selectin is augmented and prolonged in a synergistic way. The aim of the present study was to investigate if altered protein glycosylation could explain the IFN-gamma induced persistent surface expression of E-selectin. SDS-PAGE analysis of HUVEC glycoproteins, metabolically radiolabelled in the carbohydrate portion, indicated that addition of IFN-gamma produced an altered protein glycosylation. Lectin blot analysis using the Sambucus nigra agglutinin lectin also indicated differences in protein glycosylation when HUVEC were incubated with IFN-gamma/TNF-alpha compared to TNF-alpha alone. The kinetics of surface expression of E-selectin were measured using a cell ELISA assay. When HUVEC were incubated with monesin, a potent inhibitor of late Golgi function, together with both TNF-alpha and IFN-gamma, the additive effect of IFN-gamma on E selectin expression was almost abolished. Since monensin is known to affect glycosylation processing, this experiment suggested that the IFN-gamma induced change in protein glycosylation might induce the prolonged surface expression of E-selectin. However, when HUVEC were cultured with IFN-gamma/TNF-alpha in the presence of several different inhibitors of N-glycosylation processing, no significant effect of E-selectin expression was observed. Regulation of adhesion molecule expression after activation of endothelial cells is likely to play a pivotal role for the inflammatory response. Further studies are needed to understand the mechanisms underlying this regulation. PMID- 9350284 TI - Murine thymic nurse cells express ICAM-1 on caveolar and vacuolar membranes. AB - The thymic nurse cell is a unique type of epithelial cell in the thymic cortex. It is in intimate contact with the developing thymocytes by harbouring up to 200 thymocytes in distinct vacuoles, called caveoles. This investigation is concerned with the nurse cell expression of the intercellular adhesion molecule ICAM-1, the ligand for thymocyte LFA-1. Nurse cells from young Balb/c mice were isolated in a density gradient. ICAM-1 expression was studied by using two different immunotechniques: alkaline phosphatase labelled cryosections, and immunogold electron microscopy. The specific antibody was a monoclonal rat anti-mouse ICAM 1. Immunostaining of cryosections demonstrated that ICAM-1 is expressed on the surface membrane and in the internal caveolar membranes of thymic nurse cells. Electron microscopy of immunogold labelled sections revealed ICAM-1 on the surface membrane of thymic nurse cells and on the membranes of the caveoles, the small cytoplasmic vesicles, as well as on the Golgi apparatus. PMID- 9350285 TI - Regional phenotypic specialization of intraepithelial lymphocytes in the rat intestine does not depend on microbial colonization. AB - Recent studies in mice and humans have provided evidence for regional specialization of gut intraepithelial lymphocytes (IEL). Here the authors report striking regional variability in the composition of IEL in rat small and large intestine. Two-colour immunofluorescence in situ analysis showed that the distribution of the CD3+ and CD3- IEL subpopulations varied, the proportion of T cells (CD3+) being higher in the ileum than in the jejunum and smallest in the colon. These differences were explained by variable numbers of the T-cell receptor (TCR)alpha/beta + (both CD8+ and CD4+) but not the TCR gamma/delta + subset. Moreover, the various IEL subpopulations showed distinct intraepithelial distribution patterns with CD4+ and CD8 alpha beta + T cells situated near the lamina propria, while CD3- IEL were located preferentially towards the adluminal part of the epithelium. Regional phenotypic variation did not depend on intestinal colonization because analogous results were obtained in germ-free rats. Conventionalization nevertheless caused a marked relative increase of small intestinal TCR alpha/beta + but not TCR gamma/delta + IEL. This increase was more sustained in the jejunum than ileum and eventually reduced the phenotypic IEL differences between the two sites. By contrast, microbial colonization of the colon induced only a transient increase of intraepithelial TCR alpha/beta + cells with no permanent phenotypic alterations. Both CD3+ and CD3- IEL contained subpopulations that expressed NKR-P1 independent of intestinal colonization. These results demonstrate phenotypic specialization of IEL at different levels of the gut and suggest that the indigenous flora is not essential to this end. PMID- 9350286 TI - Mannuronan enhances survival of lethally irradiated mice and stimulates murine haematopoiesis in vitro. AB - Mannuronan (poly-beta-(1-->4)-D-mannuronate or poly-M), produced by Pseudomonas aeruginosa as a mucoid exopolysaccharide, has previously been shown to exhibit immunostimulating activity. The authors investigated the in vivo and in vitro effects of mannuronan on murine haematopoiesis. In vivo, prophylactic (-24 h, intraperitoneal) administration of mannuronan enhanced survival of lethally irradiated mice from zero day 40 survivors (NaCl) to 20, 80 and 70% survival at 0.5, 1 and 2 mg/kg bw mannuronan, respectively. In vitro, primary stromal cultures stimulated with mannuronan produced high levels of interleukin(IL)-1, IL 6 and colony stimulating activity. Mannuronan alone did not have any colony stimulating activity on GM-CFC, BFU-E, Mix-CFC or HPP-CFC progenitors in clonogenic assays, but acted synergistically with suboptimal amounts of growth factors on GM-CFC, Mix-CFC and HPP-CFC colony formation. Limiting dilution analysis showed that 1 of 423 bone marrow cells formed colonies in response to suboptimal GM-CSF plus mannuronan compared to 1 of 592 for suboptimal GM-CSF alone. The primitive Lin-Sca-1+ haematopoietic progenitors showed increased day 10 colony size in the presence of mannuronan in single cells assays. These stimulating effects of mannuronan on haematopoiesis may prove to have clinical importance. PMID- 9350287 TI - Complement C6 and C2 biosynthesis in syngeneic PVG/c- and PVG/c+ rat strains. AB - A PVG rat with total deficiency of C6 and partial deficiency of C2 (PVG/c-), and a syngeneic control strain (PVG/c+), were used to study the production of extrahepatically synthesized complement. Livers of complement deficient rats were transplanted in sufficient rats (Tx-L). The C6 and C2 levels in Tx-L rats declined within 2 days to 25% and 30%, respectively, and remained stable for more than 6 weeks. To investigate the contribution of C6 synthesis by the liver, C6 sufficient livers were grafted in deficient rats (Tx + 1). After an initial increase, with maximum C6 levels of 119% at 10 days following transplantation, the C6 levels decreased gradually and C6 was no longer detectable 28 days after transplantation. This decline in C6 levels was dependent on antibody production against C6. No significant change in the C3, C4, factor H and factor B levels was observed. Expression of C6 mRNA in the grafted PVG/c+ sufficient liver was comparable to the expression of C6 mRNA in control PVG/c+ livers while C6 mRNA expression in the transplanted PVG/ c- liver and the control PVG/c- liver was lower. In conclusion, it was demonstrated in vivo that not only C6 but also C2 is synthesized extrahepatically in PVG/c rats. PMID- 9350288 TI - Lymphoid cell accumulation in salivary glands of autoimmune MRL mice can be due to impaired apoptosis. AB - MRL-lpr mice and the congenic strain MRL +/+ exhibit pathological abnormalities in the salivary glands similar to Sjogren's syndrome in humans. The lpr genotype has been identified as a mutation in the gene encoding Fas which is a cell surface protein that mediates apoptosis. The mutation is leaky, allowing for low levels of the APO-1/Fas (CD95) receptor and partial activity of Fas/Fas ligand mediated programmed cell death in this strain. To examine the expression of Fas in situ, the authors analysed thymus, lymph node and salivary gland tissue from BALB/c, MRL +/+ and MRL-lpr mice by an immunohistochemical technique (ABC immunoperoxidase) using an anti-Fas (Jo2) antibody. For detection of apoptotic cells the authors used the terminal deoxynucleotidyl-transferase-mediated dUTP digoxigenin nick end labelling (TUNEL) method. Thymus from MRL +/+ and normal BALB/c mice showed a higher frequency of Fas expression than was seen in the lpr mice, but the +/+ mice had similar expression of Fas in lymph nodes as lpr mice. The Fas protein was detected among infiltrating mononuclear cells in the salivary glands of both lpr and +/+ mice. Apoptotic cells were found in the thymus with similar frequency in all three strains, while in the lymph nodes only BALB/c mice showed apoptosis. There was no, or very low, frequency of apoptosis among infiltrating mononuclear cells in salivary glands of both MRL strains. In conclusion, despite mutation of the Fas gene in the MRL-lpr strain, there was nevertheless an expression of the apoptosis-related Fas protein in lymphoid tissue and salivary glands of these mice. Based on analysis of apoptotic activity, the impaired Fas in autoimmune MRL mice seems to affect primarily the peripheral organs. PMID- 9350290 TI - Interferon-gamma production in antigen specific T cell response: quantitation of specific mRNA and secreted protein. AB - Interferon gamma (IFN-gamma) production as a measure of cellular sensitization was studied by detection of the cytokine in culture supernatant by enzyme immunoassay (EIA) and by measuring cellular mRNA using the reverse transcriptase polymerase chain reaction (RT-PCR) method. These assays were compared to the standard lymphocyte proliferation assay as a marker of T cell responsiveness to foreign antigens. When blood donors seropositive for herpes simplex virus (HSV) were compared to seronegative donors, all measurements of cellular sensitization separated the groups without overlap. There were significant correlations between the IFN-gamma mRNA titre and the secreted IFN-gamma (r = 0.57, P = 0.03), and the proliferative response and the secreted IFN-gamma (r = 0.78, P = 0.001), as well as between the IFN-gamma mRNA titre and the proliferative response (r = 0.78, P < 0.001). When tetanus toxoid (TT) responses were studied in immunized subjects, a wide range of responsiveness could be seen and correlation between various measurements was poor. However, constant individual levels of the cytokine production were demonstrated. Six people who had received their last TT booster vaccination more than 5 years ago were revaccinated and repeatedly studied. An increase in the levels of produced IFN-gamma could be seen in all subjects and two who lacked a lymphocyte proliferation response developed it after revaccination. PMID- 9350289 TI - Interferon-gamma-induced MHC class I expression and defects in Jak/Stat signalling in methylcholanthrene-induced sarcomas. AB - Seventy-eight uncloned tumour cell lines, each established from a primary sarcoma induced with methyl-cholanthrene in immunocompetent nu/+ BALB/c and C.B.-17 mice or in immunodeficient nu/nu BALB/c and severe combined immunodeficient (SCID) mice, were examined for sensitivity to interferon-gamma (IFN-gamma) as measured by tumour cell augmentation of major histocompatibility complex (MHC) class I expression. The tumour cells were cultured with IFN-gamma and their expression of Kd, Dd and Ld was measured by fluorescence-activated cell sorter analysis. All but three of the 78 tumour lines up-regulated Kd, Dd and Ld to a variable degree in response to IFN-gamma, indicating that IFN-gamma resistance is not a common property of these sarcomas. The tumour cell lines varied greatly in their MHC class I expression before as well as after IFN-gamma stimulation. There was a tendency towards a higher MHC expression after IFN-gamma stimulation in tumour lines from immunocompetent mice compared to immunodeficient mice, but no common maximum MHC class I expression level was found for the 78 tumour cell lines. Three of the tumour lines, all from immunodeficient mice, completely failed to respond to IFN-gamma by up-regulating MHC class I expression. The same three also displayed absence of IFN-gamma-induced Stat1 beta tyrosine phosphorylation and low Stat1 alpha tyrosine phosphorylation, indicating a defect in the signal transduction pathway affecting phosphorylation of Stat1. These findings strongly suggest a link between defects in Stat1 phosphorylation and the failure to up regulate MHC class I. In all tumour lines tested, the Stat1 Western blotting revealed a 78 kDa protein (p78) not previously described. PMID- 9350291 TI - Cytokine regulation of human immune response to Schistosoma mansoni: analysis of the role of IL-4, IL-5 and IL-10 on peripheral blood mononuclear cell responses. AB - The role of cytokines on the in vitro proliferative response of peripheral blood mononuclear cells (PBMC) from Schistosoma mansoni infected patients to soluble egg (SEA) and adult worm antigens (SWAP) were evaluated. The results obtained demonstrated that the proliferative response of PBMC from chronic intestinal (INT) patients to SEA and SWAP is increased by the blockage of IL-10 with specific monoclonal antibodies (MAb). The effects of these antibodies were readily reversed by the addition of recombinant IL-10. In contrast, no effect was observed on the PBMC response of acute and hepatosplenic patients (HS) in the presence of anti-IL-10. Anti-IL-4 antibodies decreased the PBMC response of the intestinal (INT) and HS individuals to SEA and SWAP, and the PBMC response of acute patients to SEA but not to SWAP. Addition of anti-IL-5 MAb did not decrease the PBMC response of acute patients to SEA or SWAP. These results suggested that IL-10 has an important role in the modulation of the immune response in chronic asymptomatic patients and that this cytokine may be an important factor in controlling morbidity. PMID- 9350292 TI - Evaluation of the role of Fc gamma and complement receptors in the decreased phagocytosis of hereditary haemochromatosis patients. AB - Hereditary haemochromatosis (HH) monocytes have a decreased antibody mediated phagocytosis of rabbit erythrocytes and Staphylococcus aureus compared to control monocytes. In order to investigate whether this decrease could be attributed to a different level of expression of Fc gamma receptors (Fc gamma R) or complement receptors (CR), which cooperate even in the absence of complement, the surface expression of these receptors was determined on monocyte-enriched suspensions. In contrast to what was expected, HH monocytes displayed a significantly higher level of Fc gamma RI and Fc gamma RIIa as compared to healthy donor monocytes, but these differences were very small. The expression of the other receptors studied were similar for both groups. The heat-inactivated mouse serum used for opsonizing the erythrocytes mainly contained mouse IgG1. Two genetically different forms of Fc gamma RIIa are known, each with a different affinity for mouse IgG1 antibodies. Therefore, the Fc gamma RIIa polymorphism in monocytes (MN) of both groups was also investigated. A similar distribution was found for patients and healthy donors. In addition, the extent of erythrophagocytosis of both donors and patients was independent of Fc gamma RIIa allotype. Our results indicate that the altered phagocytosis by HH monocytes cannot be attributed to a different level of expression of receptors involved in phagocytosis or to Fc gamma RIIa polymorphism. PMID- 9350293 TI - Interleukin-10 response abnormalities in systemic lupus erythematosus. AB - It has been previously reported that the production of interleukin-6 (IL-6) is often enhanced in systemic lupus erythematosus (SLE). The authors examined the secretion of IL-6, tumour necrosis factor-alpha (TNF-alpha), granulocyte macrophage colony-stimulating factor, IL-1 alpha and IL-4 by B cells and monocytes from lupus patients and compared this to the production in normal controls and in rheumatoid arthritis patients. IL-6 production was increased an average of 3.4-fold compared to that in normal subjects and 8.4-fold compared to rheumatoid arthritis patients. In SLE, a strongly positive correlation was found between the levels of IL-6 and TNF-alpha (R = 0.8987, P = 0.002). Since production of both IL-6 and TNF-alpha is regulated by IL-10, the enhancement of the production of these cytokines could reflect a defect in either IL-10 production or responsiveness. However, spontaneous production of IL-10 was enhanced in cultures of B cells and monocytes from lupus patients, compared to normal controls, the levels being increased 3.1- to 6-fold for monocytes and B cells, respectively. The finding of increased secretion of these cytokines implies an abnormality in IL-10-mediated suppression in SLE. To assess this possibility, the authors examined recombinant human IL-10-mediated suppression of IL-6 production by monocytes and B cells from lupus patients, compared to normal controls, and found that whereas IL-10 caused a concentration-dependent suppression of IL-6 production in normal B cells and monocytes, this suppression was deficient in B cells and monocytes from lupus patients. In SLE, it therefore appears that there may be an intrinsic defect in IL-10-induced suppression of cytokine synthesis. This could explain the increased levels of IL-10 and IL-6 found in this condition, and may also be responsible for the characteristic polyclonal B-cell activation that is seen. PMID- 9350294 TI - High levels of neutralizing autoantibodies against IL-1 alpha are associated with a better prognosis in chronic polyarthritis: a follow-up study. AB - Neutralizing autoantibodies to interleukin (IL)-1 alpha were detected in a subset of chronic polyarthritis patients characterized by an increased proportion of patients with primary Sjogren's syndrome or self-limiting inflammatory arthritis, diseases with a much better prognosis than rheumatoid arthritis (RA). The evolution of anti-IL-1 alpha antibody levels was followed over 3 years. Incidence and levels were higher in patients with a benign form of polyarthritis. In these patients levels remained stable or increased over the follow-up period. In contrast, incidence and levels were lower and some RA patients became negative. Negative correlations were observed between the levels of anti-IL-1 alpha antibodies and the clinical and biological indices of disease activity. The relative risk factor of developing RA was 12 in the absence of high anti-IL-1 alpha antibody levels and 18.2 when associated with the presence of HLA-DR4. In conclusion, the presence of anti-IL-1 alpha autoantibodies appears to be protective and their detection could represent a marker of good prognosis for destruction. PMID- 9350295 TI - Use of specific urinary proteins as diagnostic markers for renal disease. PMID- 9350296 TI - Systemic lupus: from the bedside to the laboratory bench. AB - The author highlights some of the clinical, biological and serological signs associated with systemic lupus erythematosus and use them as keys to enter into the most recent advances in the immunopathology of the disease, in particular the mechanisms underlying the production of pathogenic autoantibodies. PMID- 9350298 TI - Nosocomial bacteraemia life cost in Belgium. PMID- 9350297 TI - Beta-thalassaemia in indigenous Belgians: an update. AB - Beta-thalassaemia, a widespread autosomal recessive disorder, occurs sporadically in Northern and Western European countries. Molecular analysis of the beta-globin gene has been carried out in 30 members of 15 unrelated indigenous Belgian families which presented with non sideropenic hypochromic and microcytic anaemia. For all of them, extensive search failed to find an ancestor at risk for the disease. The beta-globin genes were first screened for frequent beta-thalassemic mutations by dot-blot hybridization with specific radiolabeled oligonucleotide probes. Direct automated fluorescence-based DNA sequencing and, in one case, Southern blotting were also used. All the 30 patients were found to be heterozygous for a beta-thalassemic mutation. Eight different mutations were identified. Among these, four are commonly found in the Mediterraneans: codon 8 ( AA), IVS-I-1 (G-->A), IVS-1-6 (T-->C) and codon 39 (C-->T); three have occasionally been reported: initiation codon (T-->C) and codon 35 (C-->A) and a rare deletion of 12.6 kb which removes all the beta-globin gene and its flanking regions. A new mutation, a -CC deletion at codon 38/39 was found in one family. These results both at the biological and molecular level show that beta thalassaemia exist in indigenous Belgian families with no known ancestor a risk for the disease. They also show that clinicians and biologists should keep in mind the existence of beta-thalassaemia in indigenous Belgian families when investigating hypochromic and microcytic anaemia in patients whom the past familial history does not evocate a risk for the disease. PMID- 9350299 TI - Phenylketonuria in Britain: genetic analysis gives a historical perspective of the disorder but will it predict the future for affected individuals? PMID- 9350301 TI - A two year prospective study to compare culture and polymerase chain reaction amplification for the detection and diagnosis of Lyme borreliosis. AB - AIM: To compare polymerase chain reaction (PCR) amplification of borrelial DNA and culture isolation of spirochaetes for the diagnosis of Lyme borreliosis by direct detection of Borrelia burgdorferi sensu lato in patients with erythema migrans and acrodermatitis chronica atrophicans lesions. METHODS: Skin biopsy specimens from erythema migrans and acrodermatitis chronica atrophicans lesions were subdivided and tested by PCR amplification assay and culture using two artificial growth media, Barbour-Stoenner-Kelly II (BSK II) and modified Kelly Pettenkofer (MKP). Five classes of lesions were studied: typical erythema migrans, spontaneously resolved erythema migrans, atypical/partially treated erythema migrans, typical acrodermatitis chronica atrophicans, and atypical/partially treated acrodermatitis chronica atrophicans. RESULTS: For both erythema migrans and acrodermatitis chronica atrophicans lesions, the most sensitive detection method was MKP culture. PCR was less sensitive than MKP culture, but more sensitive than BSK II culture. Results for 758 typical erythema migrans specimens showed positivity rates of 36% for MKP, 25% for PCR, and 24% for BSK II. Differences were statistically significant. The overall positivity rate for all three methods combined was 54%, but few specimens (6%) were positive by all three methods. Examination of multiple erythema migrans lesions from the same patient increased the diagnostic yield. These findings, and similar results for acrodermatitis chronica atrophicans lesions, suggest that the distribution of spirochaetes in skin biopsies is not homogeneous. CONCLUSIONS: Although possessing the potential to provide a rapid diagnosis, PCR is not more sensitive than culture for the direct detection of borrelia. Spirochaetes appear to be unevenly distributed throughout biopsy specimens, suggesting that diagnosis of Lyme borreliosis by direct detection of the causative agent in skin lesions in vulnerable to sample bias. PMID- 9350300 TI - Role of platelet adhesion in homeostasis and immunopathology. AB - Various molecules expressed on the surface of platelets have been shown to mediate the protective or deleterious role of these cells in immuno-inflammatory mechanisms. Increasing evidence points to the involvement of the cell adhesion molecules, gpIIb-IIIa, P-selectin, CD31, LFA-1, and CD36 in the interaction between platelets and endothelial cells as well as other cell types. The possible role of these molecules in the ability of platelets to support endothelium and to protect against tumour necrosis factor mediated cytolysis or parasitic invasion are reviewed. The involvement of platelets as effectors of tissue damage in cerebral malaria, lipopolysaccharide induced pathology, and pulmonary fibrosis is also discussed. This has then been extended to include the intercellular mechanisms underpinning their pathogenic role in metastasis, transplant rejection, stroke, brain hypoxia, and related conditions. A better understanding of the complex regulation and hierarchical organisation of these various platelet adhesion molecules may prove useful in the development of new approaches to the treatment of such diseases. PMID- 9350303 TI - Proliferative activity in human glioblastomas: evaluation of different Ki67 equivalent antibodies. AB - AIMS: Determination of proliferative activity in human gliomas may be of clinical importance. Immunohistochemical estimation of the proliferative index with the prototypic monoclonal antibody Ki67 is often used but has the disadvantage that it must be carried out on frozen material. However, novel Ki67 equivalent antibodies have been developed for use on formalin fixed and paraffin wax embedded tissue. In this study, the prototypic Ki67 antibody and several new Ki67 equivalent antibodies were tested on human glioblastoma tissue. METHODS: Eleven glioblastomas were included in the study. The antibodies used were the prototypic monoclonal Ki67 and the novel Ki67 equivalent antibodies MIB1 (monoclonal), NC MM1 (monoclonal), NC-Ki67p (polyclonal), and rabbit antihuman Ki67 antigen (polyclonal). The prototypic Ki67 was used on frozen sections and the other Ki67 antibodies on microwave oven heated, formalin fixed and paraffin wax embedded sections. RESULTS: All antibodies exhibited specific granular nuclear staining of weak to strong intensity. In some tumours the labelling indices were within the same range, whereas in others the antibodies elicited divergent values. CONCLUSIONS: All the novel Ki67 equivalent antibodies provided satisfactory staining on paraffin sections. However, a significant spread of labelling indices was recorded in some cases. Therefore, Ki67 immunostaining is encumbered with some degree of uncertainty and requires further optimisation before it can be regarded as a reliable prognostic marker. PMID- 9350302 TI - Mature osteoblasts in human non-union fractures express collagen type III. AB - AIMS: High levels of collagen type III are biochemically detectable in biopsies of non-uniting fractures, and in the serum of patients suffering from this condition. The aim of this study was to determine whether the expression of collagen type III was limited to fibrous tissue in non-unions, or whether some was present in bone. METHODS: Biopsies from normally healing human fractures and non-unions were examined using in situ hybridisation and immunohistochemistry. RESULTS: The mesenchymal cell population, which includes fibroblast and osteoblast precursors, expressed mRNA for collagen type III. However, mature osteoblasts on the surface of woven bone varied profoundly between normally healing fractures (in which they were negative or occasionally weakly positive) and non-unions (in which they were strongly positive). Areas of woven bone that had osteoblasts positive for collagen type III mRNA also immunostained positively for the protein. CONCLUSIONS: This study shows that non-union fracture callus osteoblasts on the surfaces of woven bone exhibit an unusual phenotype: they express collagen type III, a molecule characteristic of an earlier stage of osteoblast differentiation, which is not expressed by osteoblasts on woven bone surfaces of bone that develops normally. This finding may be useful in developing an early clinical test for impending non-union. PMID- 9350304 TI - NO38 expression and nucleolar counts are correlated with cellular DNA content but not with proliferation parameters in colorectal carcinomas. AB - AIMS: To investigate the expression of nucleolar protein NO38, to determine the numbers of nucleoli per cell, and to examine the relations of these nucleolar parameters to tumour DNA index, total cellular DNA content, S phase fraction, and Ki67 labelling index. METHODS: 36 colorectal tumours and 14 normal mucosas were studied. An anti-NO38 monoclonal antibody, 31A12, and flow cytometric analysis were used to detect expression of NO38 by means of a biotin-streptavidin-FITC (fluorescein isothiocyanate) staining method. Nucleolar counts were determined using fluorescence microscopy. Flow cytometry was used to determine tumour DNA indices and the sizes of the S phase fractions. Ki67 labelling indices were determined from tissue sections stained immunohistochemically with the MIB-1 antibody against the Ki67 nuclear protein. RESULTS: Generally, tumour cell nucleoli were larger and more irregular in shape compared with nucleoli in normal mucosal cells. DNA aneuploid and diploid tumours expressed 2.8 and 2.1 times more NO38 than normal mucosa. The mean (SD) values for nucleolar counts were higher for the DNA aneuploid tumours (3.81 (0.93)) than the diploid tumours (2.62 (0.38)) and normal mucosa (2.34 (0.37)). NO38 expression and numbers of nucleoli correlated significantly (r = 0.52, p = 0.01). There were, however, no significant correlations between these nucleolar parameters and either the sizes of tumour S phase fractions or Ki67 labelling indices. Cell cycle resolved expression of NO38 in tumours and normal mucosa demonstrated that expression increased approximately in proportion to the DNA content throughout the cell cycle. In aneuploid tumours, NO38 expression was 43% and 98% higher in S and G2 phases, respectively, compared with the G1 phase. Sorting of these populations revealed that the nucleolar count also increased as the DNA content increased but by only 29% and 47% in S and G2, respectively. Apoptotic cells lacked NO38. CONCLUSIONS: NO38 expression is higher in tumours than in normal mucosa owing to the increased DNA content and larger nucleoli in tumours; expression increases proportionally with DNA content as cells progress through the cell cycle from G1 through S and G2. However, NO38 expression does not correlate with the tumour S phase fraction or Ki67 labelling index and is lost during apoptosis. Also the results suggest that nucleoli grow in size during the cell cycle, which would account for the doubling of NO38 expression from G1 to G2, as the nucleolar count increased by only 47%. PMID- 9350305 TI - Reverse transcriptase-polymerase chain reaction analysis of cytokeratin 19 expression in the peripheral blood mononuclear cells of normal female blood donors. AB - BACKGROUND: Early detection of haematogenous dissemination of epithelial tumours afforded by the analysis of epithelial antigen expression in the peripheral blood mononuclear fraction (PBMN) and bone marrow may confer a worse prognosis to patients with carcinoma. Cytokeratin 19 is a protein normally expressed by epithelial cells including normal and malignant mammary cells. Previous studies have demonstrated that analysis of cytokeratin 19 expression by the reverse transcriptase-polymerase chain reaction (RT-PCR) can detect one epithelial cell in as many as 10(5)-10(7) haematopoetic cells. Despite its sensitivity concern has been voiced recently about the specificity of this technique owing to the detection of cytokeratin 19 expression in the PBMN of normal volunteers and the bone marrow of patients with haematological malignancies. AIMS: To assess the sensitivity and specificity of RT-PCR detection of cytokeratin 19 in PBMN of normal female blood donors. METHODS: Blood was taken from 52 normal female blood donors and PBMN separated through Fycol gradient centrifugation. Cytokeratin 19 was measured using a two step nested RT-PCR assay. RESULTS: No amplification was found in the first step for any of the samples studied, whereas in the second step amplification was observed in 10 of the 52 samples. Both steps could detect one MCF-7 cell (the cytokeratin 19 positive control) in 10(6) CEM (cytokeratin 19 negative control) cells. CONCLUSIONS: As both PCR steps are sensitive to the 10( 6) level, performing only the first amplification step may decrease the non specificity of this method. Further studies are needed to define the specificity and sensitivity of this technique in blood and bone marrow specimens of women with breast cancer. PMID- 9350307 TI - Microdissection of stained archival tissue. AB - In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns sections are dewaxed, stained with haematoxylin and eosin, dried, covered with Sellotape, and the tissue cut out using a scalpel blade under direct visual control. The method is quick, eliminates problems of operator tremor, preserves the architecture of the micro-dissected tissue (for photographic documentation) and requires no special equipment. The presence of Sellotape and adhesive in the reaction mixture has no detrimental effect on the ability to extract DNA or to perform PCR. PMID- 9350306 TI - Fatty acid oxidation disorders as primary cause of sudden and unexpected death in infants and young children: an investigation performed on cultured fibroblasts from 79 children who died aged between 0-4 years. AB - BACKGROUND: Disorders of fatty acid metabolism are known to be responsible for cases of sudden and unexpected death in infancy. At least 14 disorders are known at present. 120 cases of sudden infant death syndrome (SIDS) had been examined for a prevalent mutation (G985) causing medium chain acyl CoA dehydrogenase deficiency, which is inherited in an autosomal recessive mode. No over representation of either homozygous or heterozygous cases was found. AIMS: To investigate a broader spectrum of fatty acid oxidation disorders in a wider range of sudden deaths in infants and young children. METHODS: Seventy nine cases of unexpected death in infants and young children younger than 4 years old were examined for a minimum of nine fatty acid oxidation disorders, using the global [9, 10-3H] myristic acid oxidation assay in cultured fibroblasts from achilles tendon biopsies taken at postmortem examination. RESULTS: Three cases with fatty acid oxidation disorders and two carriers of the G985 mutation were found, all categorized as non-SIDS or borderline SIDS. The global assay used has the advantage of simplicity. CONCLUSIONS: These results indicate that disorders of fatty acid oxidation play a small but significant role in the cause of unexpected death in infants and young children, and that infants and children dying in this way should be regarded as high risk candidates for metabolic diseases. PMID- 9350308 TI - A new mutation in the human lipoprotein lipase gene causing familial hyperchylomicronaemia. AB - Lipoprotein lipase plays a major role in the regulation of lipid metabolism. The enzyme acts to hydrolyse triglycerides, providing free fatty acids for energy generation or storage, thus affecting the maturation of circulating lipoproteins. Biochemical and molecular analyses were performed on two siblings of consanguineous Pakistani origin, presenting with hyperchylomicronaemia, which revealed that the disorder resulted from lipoprotein lipase deficiency. Molecular analysis of the lipoprotein lipase gene has revealed a novel homozygous mutation, leucine to proline at amino acid residue 303, within the amino terminal domain of the protein. PMID- 9350309 TI - Human papilloma virus detection by in situ hybridisation signal amplification based on biotinylated tyramine deposition. PMID- 9350310 TI - Evaluation of proliferating cell nuclear antigen (PCNA) in supraglottic carcinoma. AB - It is widely accepted that tumor invasion and metastasis are the primary causes for the lethal clinical outcome of cancers. In recent years, attention has been drawn to PCNA (Proliferating Cell Nuclear Antigen) to predict the prognosis of some cancers. In the present paper, we have studied anti-PCNA antibody PC10 in supraglottic cancer by means of immunohistochemistry using paraffin-embedded sections, to demonstrate its clinical significance in this type of malignancy. Twenty-two patients with supraglottic cancer, including T1 (5 cases), T2 (13 cases) and T4 (4 cases) with N- (14 cases) and N+ (8 cases), were investigated for the PCNA expression. The percentages of PCNA positive cells were divided into three groups: < 25%, 25-75%, > 75%, with the range from 10.6 to 95.2%. Results showed that PCNA was well correlated with lymph node metastasis, and appeared to have an inverse correlation with histopathological grades. In this small group, we did not find that PCNA was correlated with T stages and tumor size. However, compared with other T-stages and tumor sizes, the correlation between lymph node metastasis and PCNA seemed to have more clinical significance in T2-stage and in tumors larger than 2 cm. PCNA could be used as a marker in predicting the clinical outcome in supraglottic cancers. An analysis on a large scale is anticipated. PMID- 9350311 TI - Temporary voice changes after uncomplicated thyroidectomy. AB - Voice characteristics were studied before and after thyroidectomy in patients with intact vocal fold motility. The speaking voice was acoustically analysed in 47 patients and phonetograms were made in 17 patients. Eight parameters were measured and the pre- and postoperative values compared. The results show that the most affected parameter was the pitch of the speaking voice. The fourth postoperative day there was, on average, a lower SFo and a smaller Fo range during speaking. Postoperatively a progressive normalisation took place. After three months there were no more statistical differences and, looking at the individual measures, the SF0 of all patients fell within 2 semitones from their preoperative level. Vocal quality was also altered in the first postoperative examination, as shown by the higher jitter and smaller harmonics. These measures normalised after two weeks. In the same way, the evaluation of the limits of the voice by means of the phonetogram, showed that the maximal performances in the intensity and pitch domain were decreased in the earliest postoperative period. Information about temporary voice change is useful in patients undergoing thyroidectomy. PMID- 9350313 TI - Hypernasality as a strategy in speech sound acquisition: a case report. AB - Hypernasality is a common problem in children that may have various causes, organic as well as functional ones. This paper reports a case of hypernasality in a 5-year-old girl. The speech pattern she displayed was unusual in that the hypernasality apparently functioned as a strategy in speech sound acquisition. PMID- 9350312 TI - Subjective results after uvulo-palato-pharyngoplasty. AB - The subjective effect of uvulo-palato-pharyngoplasty surgery (U.P.P.P) for snoring was evaluated in 31 patients. Results were obtained following a questionnaire sent to patients after a median follow-up of 10 months, as well as through follow-up visits. Seventy seven point five percent of the patients and their partners were satisfied with the operation. No major complications were noted. Minor complications included pain, transient pharyngonasal reflux and sensitvity problems of the posterior wall of the pharynx. PMID- 9350314 TI - Acoustical control of voice changes after surgery: an exploratory essay. AB - In this preliminary study, the authors try to validate the use of a quantitative method based upon the use of Long Term Average Spectra (LTAS) and indices of spectral (dis)similarity. The whole spectral information is used and an evolution ratio is introduced. The technique has been tested on 5 patients with organic lesions of the vocal cords (oedema, polypoid degeneration, hyperkeratosis, polyp, pseudocyst). Each patient has been recorded three times: the day before surgery, one and 5 weeks later. The obtained values suggest good correlation between the index variability and the magnitude of clinical change. The values obtained from the quantitative technique turn out to be well correlated with the evolution of the patients' state. PMID- 9350315 TI - Nasal submucosal glands: lectin histochemistry study. AB - Submucosal glands (SG) are the most important secretory elements in the human nasal mucosa. However, there is little information about the glycoconjugates of the SG secretions. Ten patients were selected in an aleatory way and a wide biopsy of the right inferior nasal turbinate mucosa was performed. The patients did not present any otorhinolaryngologic disease and the nasal exploration was normal. The serous cells of SG show a moderate reactivity to WGA, N-PNA, N-WGA and ConA and mild to WEA-I. The mucus cells of SGNM demonstrate a strong reactivity to N-PNA and N-WGA, moderate to WEA-I and WGA and there is no reactivity to Con-A. For DBA, SBA and LTA lectins no reactivity was observed. PMID- 9350316 TI - Posterior epistaxis: our experience with transantral ligation and embolisation. AB - Transantral ligation of the arteria maxillares internae (AMI) (with or without ligation of the arteria ethmoidalis) and percutaneous embolisation of AMI in cases of intractable epistaxis (i.e. epistaxis not responding to classical posterior packing) are compared. Except when ethmoidal bleeding is suspected, we recommend embolisation for epistaxis not responding to classical measures as it leads to fewer complications and recurrences, to shorter hospital stay and to improved postoperative comfort. PMID- 9350317 TI - Conservative treatment of chronic maxillary sinusitis in children. Long-term follow-up. AB - A placebo-controlled prospective study is performed to evaluate conservative treatment of chronic maxillary sinusitis in children. Antibiotic treatment and drainage do not seem to have a permanent curative effect. PMID- 9350318 TI - Unusual choanal polyps. AB - Four children with choanal polyps are presented. Their age and/or the site of origin of the polyps are unusual. At the time of diagnosis, the two boys were respectively 3 years 9 months and 4 years 6 months. The two girls were aged 9 and 10. In one of the patients the polyp originated from the sphenoidal sinus, in another from the inferior turbinate. The age of the child complicates diagnosis, endoscopic sinus surgery and postoperative care. PMID- 9350319 TI - Morpho-functional partition of the middle ear cleft. AB - The observation of an inter-attico-tympanic diaphragm in the middle ear (ME) cleft and the functional histology of the mucosa lead to a concept of ME morpho functional partition. There are two separate compartments: an antero-inferior one, principally devoted to the muco-ciliary clearance function, and a postero superior one, more devoted to the gas exchange function. With this concept of partition the mechanisms involved in the pathogeny of "otitis media" can be better understood. PMID- 9350320 TI - Large vestibular aqueduct syndrome with mixed hearing loss: a case report. AB - A case report of a large vestibular aqueduct syndrome (LVAS) as the sole radiographically detectable inner ear anomaly with a mixed hearing loss is presented. Hearing loss in LVAS is sensorineural with a conductive component in 15-27% of the cases. The hearing loss fluctuates and is progressive, resulting in the deterioration of the hearing after minor head trauma. The pathophysiology of the sensorineural hearing loss and the conductive component remains unknown. Endolymphatic sac surgery has been abandoned because of possible adverse results. In our case a complex anomaly was found in the stapes region without noticeable hearing gain after correction. In case of a progressive mixed hearing loss in children with a normal tympanic membrane, a CT-scan is mandatory to rule out LVAS before attempting middle ear surgery that is to no avail. PMID- 9350321 TI - Osteoma of the internal auditory canal. AB - A case of an osteoma of the internal auditory canal in a patient with severe hearing loss is presented. Osteomata of the internal auditory canal have only rarely been diagnosed. Symptoms consist in sensorineural hearing loss, vertigo and/or tinnitus. CT scan is an appropriate imaging modality. In selected cases surgery can be beneficial. A review of the literature is presented. PMID- 9350322 TI - The relevance of using tragal cartilage in tympanoplasty. AB - Chronic endotympanic depression is a pathological situation which leads to tympanic atelectasis, retraction pockets and cholesteatoma. It is also at the origin of tympanoplasty failure. Tragal composite perichondrium-cartilage graft is a procedure which gives good results in these difficult surgical cases. PMID- 9350323 TI - Pregnancy following intracytoplasmic sperm injection of immotile spermatozoa selected by the hypo-osmotic swelling-test: a case report. AB - The success of intracytoplasmic sperm injection (ICSI) is poor when only immotile spermatozoa can be retrieved. In a couple with complete male asthenozoospermia the possible use of the hypo-osmotic swelling test to select spermatozoa for microinjection was examined. Following incubation in hypo-osmotic medium (Hypo 10, IVF Science, Goteborg, Sweden), 26% of immotile spermatozoa showed signs of sperm swelling (HOS-positive). After injection of HOS-positive spermatozoa, 5 out of 12 oocytes fertilized (41%) and after transfer of three embryos a healthy singleton pregnancy was achieved. In a previous ICSI cycle of this couple without preselection of spermatozoa by the HOS test, only 1 out of 10 oocytes fertilized. It is concluded that selection of spermatozoa by hypo-osmotic swelling-test prior to sperm microinjection seems to be a valuable tool to increase the fertilization rate in cases with complete asthenozoospermia. PMID- 9350325 TI - Localization of adhesion molecules on human spermatozoa by fluorescence microscopy. AB - The expression of adhesion molecules on human spermatozoa of healthy probands was analysed. The localization patterns of adhesion molecules (AM) on the spermatozoal surface were documented by fluorescence microscopy. Spermatozoa were incubated with antibodies against alpha 1 (CD49a), alpha 2 (CD49b), alpha 3 (CD49c), alpha 4 (CD49d), alpha 5 (CD49e), alpha 6 (CD49f) chains of beta 1 integrins, beta 1 (CD29), beta 2 (CD18), alpha V (CD51), beta 3 (CD61) and beta 4 integrin chains, the LFA-3 (Lymphocyte function antigen, CD58) from the immunoglobulin superfamily and the extra-cellular matrix proteins laminin, fibronectin and collagen IV. For collagen IV, alpha 1 and alpha 2 chains no expression could be noticed. Laminin was detected at the acrosomal membrane, fibronectin and beta 4 chain mainly at the equatorial membrane. The fibronectin receptors alpha 3, alpha 4 and alpha 5 chains of the beta 1 integrins were mainly located on acrosomal and equatorial membrane areas. Laminin receptor alpha 6 chain was located postacrosomal and less frequently acrosomal. beta 2 chain and vitronectin receptors alpha V and beta 3 chains had a mainly postacrosomal localization pattern. LFA-3 was found constantly on postacrosomal membrane areas. Double staining technique was used to prove the simultaneous occurrence of fibronectin and its integrin receptors alpha 3, alpha 4 and alpha 5 chains and of alpha V and beta 2 chains on spermatozoa. The localization patterns of integrins on double stained spermatozoa were similar to the patterns described for single stained spermatozoa. The localization of fibronectin appeared to be influenced by the presence of integrins: the typical equatorial fibronectin band disappeared in case of an equatorial localization of integrins. PMID- 9350324 TI - Calcium requirement and time course of capacitation of goat spermatozoa assessed by chlortetracycline assay. AB - We standardized chlortetracycline fluorescent assay for studies of calcium requirement and time course of capacitation of goat spermatozoa. Three distinct fluorescent patterns were easily detected in goat spermatozoa incubated under capacitating conditions. Categorised according to nomenclature reported earlier, these are: 'F' with bright fluorescence in the postacrosomal region, characteristic of uncapacitated acrosomal-intact cells; 'B' with bright fluorescence on the anterior portion of the head and dark band in the postacrosomal region, characteristic of capacitated, acrosome-intact cells; 'AR' with lack of fluorescence on the head characteristic of acrosome-reacted cells. A close correspondence was observed when the results of CTC assay were compared with those obtained by transmission electron microscopy. Goat spermatozoa were not capacitated when calcium was omitted from the medium and 80% had CTC fluorescence of 'F' type. The size of 'B' cell population increased with increase in calcium concentration; at 1.0 mmol l-1 a peak representing 65-70% capacitated cells accumulated in 4 h. At higher concentrations, 'AR' cells were found along with 'B' cells and the two cell types were in equal proportions at 1.71 mmol l-1. Time course studies revealed a 2 h incubation period at 1.0 mmol l-1 and 1 h at 2 mmol l-1 calcium concentration before transformation of 'F' cells to 'B' cells was noticed. However, at no time were 'AR' cells found exclusively pointing to an equilibrium between the two sperm populations. Goat spermatozoa were also not capacitated when phosphate was omitted from the medium. Permeant anions (NO3-, SCN-), permeant weak acid (HCO3-) and organic phosphates (beta-glycerophosphate, glucose-6-phosphate) were unable to replace phosphate. The reason for their failure for the incidence of capacitation was traced to low uptake of calcium by goat spermatozoa. In the presence of phosphate, a 6-8-fold increase was measured over the calcium uptake when phosphate was omitted (2-4 nmol l-1 10(8) cells-1). Mersalyl inhibited the calcium uptake by goat spermatozoa as well as its capacitation most likely by inhibiting the calcium phosphate transporter located in the sperm plasma membrane. PMID- 9350326 TI - Monoclonal antibodies to canine intra-acrosomal sperm proteins recognizing acrosomal status during capacitation and acrosome reaction. AB - Monoclonal antibodies Ds-1 and Ds-2 specifically labelling dog sperm acrosome were prepared by immunization of mice with acetic acid extracts of dog spermatozoa. Electron microscopy and indirect immunofluorescence localized the site of Ds-1 and Ds-2 proteins inside the acrosomal vesicle. Ds-1 antibody detected 55, 76, 115, 120 and 190 kDa proteins under non-reducing conditions, and 73 kDa and 54 kDa proteins after reduction (p73/Ds-1 and p54/Ds-1). 92 kDa and 40 kDa proteins recognized by Ds-2 (p92/Ds-2 and p40/Ds-2) migrated at > 200 kDa in the absence of reducing agent. In vivo, p73/Ds-1 and p54/Ds-1 are therefore likely to be present both in free and complexed form, while all of p92/Ds-2 and p40/Ds-2 form disulfide-bonded complexes. Decrease in the rate of acrosomes stained with Ds-1 and Ds-2 was correlated with the progress of capacitation resulting in the increased rate of spontaneous acrosome reactions, as suggested by a dramatic effect of A23187. Monoclonal antibody to boar acrosin (ACR-2) recognized dog sperm acrosin homologue. A higher rate of ACR-2-negative spermatozoa was observed after capacitation and A23187 treatment compared to Ds-1 and Ds-2, indicating that proteins recognized by Ds-1 and Ds-2 are localized in a specific compartment of acrosome, distinct from acrosin and possibly representing fraction of acrosomal matrix. PMID- 9350327 TI - Effect of natural antioxidants, superoxide dismutase and hydrogen peroxide on capacitation of frozen-thawed bull spermatozoa. AB - Bovine spermatozoa from frozen-thawed semen are sensitive to lipid peroxidation. Vitamin E protects sperm membrane against oxidative damage. Sperm capacitation produces structural changes on the plasma membrane. Reactive oxygen species could be involved in the capacitation process. The aim of this work was to study the influence of natural antioxidants on the plasma membrane and the influence of reactive oxygen species during bovine sperm capacitation. Sperm samples were frozen in a standard diluent, with and without vitamin E (1 mg ml-1). Heparin (60 micrograms ml-1) was used as a sperm capacitation inductor. Sperm capacitation was evaluated by chlorotetracycline assay. Lipid peroxidation was determined by the 2-thiobarbituric acid assay. A diminution of thiobarbituric acid reactive substances was observed in sperm samples frozen with vitamin E (P < 0.05). The addition of vitamin E to the freezing diluent had no effect on the capacitated pattern (P > 0.05). When vitamin E and vitamin E + vitamin C were added to the capacitation medium, a significant decrease in the percentage of capacitated spermatozoa (P < 0.05) was observed in both cases. The addition of superoxide dismutase (0.1 mg ml-1) or H2O2 (50 microM) in the incubation medium, decreased the percentage of capacitated spermatozoa (P < 0.05). Vitamin E protects the plasma membrane against lipid peroxidation during sperm capacitation, and the presence of superoxide anion would be necessary for frozen-thawed bull sperm capacitation. PMID- 9350328 TI - Notulae seminologicae. 8. Ultrastructural sperm defects in two men, carriers of autosomal inversion. AB - The electron microscopical analysis of spermatozoa in two infertile male carriers of a pericentric inversion in one of the chromosomes 9 revealed the presence of a peculiar defect affecting the tails' fibrous sheath in both patients. This structure appeared completely disorganized and hyperplasic; sometimes the defect was associated with other usual malformations concerning the nuclear and acrosomal shape and texture and the axonemal assembly. Most spermatozoa (90-100%) of these patients were immotile. Our findings point to a definite ultrastructural sperm defect found in cases of autosomal inversion. PMID- 9350329 TI - The junctions between efferent ductules and epididymal duct in the boar. AB - Sperm transit through the male excurrent duct system is dependent on complete luminal patency. Since male excurrent ducts are derived embryologically from originally separate structures, the junctions between mesonephric tubules and the mesonephric duct are of major interest with respect to obstructive disorders. Therefore, we investigated the junctions between efferent ductules and epididymal duct in adult boars by means of serial semithin and ultrathin sections. Based on the anastomosing pattern and on the site of transition from low columnar, ciliated epithelium to high columnar, non-ciliated epithelium, three different types of anastomoses were identified, one of which was associated with luminal stenosis and sperm accumulation. The occurrence of end-to-side junctions with the epithelial transition being localized within the lumen of the tributary may thus impede normal sperm transport. PMID- 9350330 TI - Cationic and secretory effects of 6-O-acetyl-D-glucose in rat pancreatic islets. AB - Selected esters of D-glucose were recently proposed as tools to supply the sugar to cells affected by a defect in the carrier-mediated process of hexose transport across the plasma membrane. The present study extends this knowledge to 6-O acetyl-D-glucose. At a 2.0 mM concentration, the ester was indeed found to stimulate insulin release from perifused rat pancreatic islets. This coincided with stimulation of calcium influx into the islet cells, as judged from comparison of the ester effects on 45Ca outflow from prelabelled islets perifused in either the absence or presence of extracellular Ca2+. The facilitated inflow of Ca2+ did not appear attributable to a decrease in K+ conductance, 6-O-acetyl-D glucose augmenting 86Rb efflux from islets prelabelled with this radioactive cation. Both the latter phenomenon and the insulinotropic action of 6-O-acetyl-D glucose failed to be antagonized by D-mannoheptulose. These findings indicate that the cationic events coupling the recognition of D-glucose to the stimulation of insulin release are not identical in islets exposed to the hexose or its ester. PMID- 9350331 TI - Purification and characterization of an endoglucanase from the marine rotifer, Brachionus plicatilis. AB - The marine rotifer, Brachionus plicatilis, is able to digest Chlorella efficiently, suggesting that the rotifer contains a powerful cellulolytic enzyme system. A multi-component cellulolytic complex, including endoglucanase (CM cellulase), cellobiohydrolase and beta-glucosidase, was found in Brachionus plicatilis. Endoglucanase (endo-beta-1,4 glucanase) was purified to homogeneity from rotifer homogenates using a sequential chromatographic method. The purified enzyme exhibits a strong hydrolytic activity with carboxymethyl(CM)-cellulose. The optimum temperature and pH for the endoglucanase activity were 37 degrees C and 7.0, respectively. 80% of the CM-cellulase activity was retained in salt mixture that ranged from 150 to 500 mM NaCl equivalent. The purified protein was isolated with a molecular weight of approximately 62 kDa estimated by SDS polyacrylamide gel electrophoresis. PMID- 9350332 TI - Functional analysis of the evolutionary conserved arginine 182 residue in human glutathione S-transferase P1-1. AB - The mutational replacement of Arg182 with threonine markedly decreased the specific activities for GSH-conjugation reaction. The Kcat of R182T for GSH-[1 chloro-2,4-dinitrobenzene] conjugation reaction was about 100-fold smaller than that of the wild type. On the other hand, the affinity for GSH of R182T was not significantly affected. The pKa of the thiol group of GSH bound in R182T was approximately 0.9 pK units higher than those in the wild type, but the Kcat/Km1 chloro-2,4-dinitrobenzene values at high pH were not so much lower than those of the wild type. The thermostability of R182T was significantly lower than that of the wild type. Therefore, Arg182 seems to be important for the construction of the active enzyme structure. PMID- 9350333 TI - Effect of 4-methylumbelliferone on cell-free synthesis of hyaluronic acid. AB - Human skin fibroblasts were cultured in the presence of 0.5 mM 4 methylumbelliferone for 12 h, and cell-free synthesis of hyaluronic acid was performed using membrane-rich fraction from the cells. The preincubation of the cells with 4-methylumbelliferone reduced hyaluronic acid synthesis to 15% of that of non-preincubated cells, although its chain length was not changed. On the other hand, without preincubation of the cells with 4-methylumbelliferone, hyaluronic acid synthesis was not changed even when 4-methylumbelliferone was added directly to the reaction mixture. These results suggest that 4 methylumbelliferone represses the expression of hyaluronic acid synthase on the cell surface. PMID- 9350334 TI - Levels of interleukin-6, CRP and alpha 2 macroglobulin in cerebrospinal fluid (CSF) and serum as indicator of blood-CSF barrier damage. AB - We measured the levels of interleukin-6 (IL-6), albumin, C-reactive protein (CRP) and alpha 2 macroglobulin (alpha 2M), all of which have different spectrums of molecular weight, in the cerebrospinal fluid (CSF) and serum in 121 patients to evaluate damage to the blood-cerebrospinal fluid barrier (BCB) in meningitis. There was an extraordinary high level of IL-6 in the CSF when patients had bacterial or viral meningitis, but the level returned to a normal range within a week in almost all of these cases. There were no significant differences in CSF albumin levels among the different disease groups. The CRP level in CSF is considered to correlate with the serum level, and CSF CRP was higher in bacterial meningitis than in viral meningitis, however, CRP in CSF was increased in some of the infectious diseases without meningitis. The alpha 2M in CSF, which tends to be at extraordinarily high levels when there is damage to the BCB, correlated highly with CSF cell counts. CSF IL-6 seemed to be a useful indicator to identify the acute active phase of meningitis. CRP and alpha 2M in CSF are considered to be useful to differentiate bacterial meningitis, bacterial infection without meningitis and viral meningitis. Extraordinarily high levels of alpha 2M, which has a high molecular weight, in CSF is indicative of BCB damage. PMID- 9350335 TI - High affinity bradykinin binding to human inflammatory cells. AB - Specific direct bradykinin (BK) binding and competitive inhibition was detected in human neutrophil and peripheral blood mononuclear cell (PBMC) detergent solubilized extracts and purified plasma membranes using in vitro radioreceptor ligand binding. Scatchard analyses of [125I]-BK binding revealed an equilibrium dissociation constant (Kd) of 2.9 x 10(-11) M for neutrophils and 5.6 x 10(-11) M for PBMC using [des-arg9]-BK a B1 agonist; 2.6 x 10(-11) M for neutrophils, 6.2 x 10(-11) M for PBMC with BK a B2 agonist; 5.4 x 10(-11) M for PBMC using Lys-BK a B2 agonist. The number of binding sites (Bmax) was calculated to be 0.113 fM/microgram protein (720 receptors per cell) for neutrophils and 0.200 fM/microgram protein (1289 receptors per cell) for PBMC with the B1 agonist while with the B2 agonists the values were 0.128 fM/microgram protein (818 receptors per cell) for neutrophils and 0.157 fM/microgram protein (1005 receptors per cell) for PBMC with BK, and 0.293 fM/microgram protein (1870 receptors per cell) with Lys-BK for PBMC. In a competitive binding inhibition assay using neutrophil and PBMC glycerol purified plasma membranes, high affinity binding in the nanomolar range was detected to Lys-BK and BK but with [des-arg9]-BK a 10-100 fold lower order affinity was observed this being indicative of pharmacologically defined B2 characteristics. PMID- 9350336 TI - Hydroxyl radical-induced decline in motility and increase in lipid peroxidation and DNA modification in human sperm. AB - We employed the xanthine-xanthine oxidase system to produce H2O2 or simply used commercially available H2O2 solution to investigate the effects of exogenous hydroxyl radicals on the motility characteristics and on lipid peroxidation and DNA modification of human sperm. The functional parameters of sperm motility declined concomitantly upon incubation of sperm with hydroxyl radicals. After incubation of freshly ejaculated human sperm with 0.23 mM H2O2 in the presence of 1.8 mM ADP and 2.7 mM FeSO4 for 1 hr at 37 degrees C, 90% reduction of motility was observed. Effect of hydroxyl radicals on sperm motility was dependent on the concentrations of FeSO4 and H2O2, respectively. The remaining motility of sperm after 1 hr incubation showed negative linear correlation with FeSO4 concentration. The response of sperm motility to FeSO4 was also dependent on the concentration of H2O2. Except for the amplitude of lateral head displacement, functional parameters of sperm declined with the increase of H2O2 concentration. Moreover, we found that lipid peroxidation measured as malondialdehyde (MDA) and accumulation of modified DNA indicated by 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in human sperm were significantly accelerated by exogenous hydroxyl radicals. The contents of lipid peroxides and 8-OH-dG in the spermatozoa were increased from 24.6 +/- 2.4 nmol MDA/1 x 10(7) sperm and 0.17 +/- 0.02% in the untreated group to 30.6 +/- 1.2 nmol MDA/1 x 10(7) sperm and 1.9 +/- 0.47%, respectively, in the sperm treated at 37 degrees C for 1 hr with 2.03 mM H2O2, 1.8 mM ADP and 4.5 mM FeSO4. Taken together, these results suggest that the detrimental effects of hydroxyl radicals on human sperm functions may be mediated, at least partly, through lipid peroxidation and DNA modification. PMID- 9350337 TI - Iron as a possible mediator of the oxic-to-anoxic transition in Paracoccus denitrificans. AB - Expression of the lacZ gene from the Fnr-regulated FF-melR promoter on a plasmid in iron-deprived Paracoccus denitrificans cells required not only a decreased oxygen tension but also supplementation with iron. The levels of beta galactosidase and 5-aminolevulinate synthase showed comparable responses to changes in iron availability. The presence of soluble and particulate enzymes catalyzing the reduction of Fe(III) by NADH suggests a hypothesis in which the redox state of the cytoplasmic NAD-couple determines the concentration of free Fe(II) and thereby modulates the activity of a common regulator of the Fnr type. PMID- 9350338 TI - Inhibition of platelet aggregation by L-arginine and polyamines in alloxan treated rats. AB - The antiaggregating effect of L-arginine and polyamines (putrescine, spermidine and spermine) was studied in platelets of normal and diabetic rats (120 mg/Kg alloxan, i.p.). This effect was compared with insulin. The assays of platelet aggregation were carried out using platelet-rich plasma (PRP) obtained from both groups of animals. The platelet aggregation test were first standardized by using PRP obtained from human health donors. 2.5 mumoles/ml ADP, 250 mumoles/ml epinephrine and 0.4 U/ml of thrombin were used as inductors of platelet aggregation. 60% of inhibition was observed with 10 microM of L-arginine or polyamines in PRP of normal rats, and 50% with PRP of diabetic rats when thrombin was used as an agonist. These results show that L-arginine and the polyamines putrescine, spermidine and spermine have an antagonist action in platelet aggregation. In addition, we demonstrated the platelet arginase activity not only in rat platelets but also in human, was less under hyperglycaemia. The activity of this enzyme has been associated with polyamine synthesis, required to regulate platelet function. PMID- 9350339 TI - Penetration of erythrocyte membrane by peroxynitrite: participation of the anion exchange protein. AB - Participation of the anion exchange protein (Band 3 protein) of the erythrocyte membrane in the transport of peroxynitrite into erythrocytes was shown by partial inhibition of hemoglobin oxidation by extracellular peroxynitrite in cells treated with Band 3 inhibitors. These results demonstrate that permeation in the anionic form may be a minor pathway in the membrane transport of the peroxynitrite anion/peroxynitrous acid couple, especially in membranes reach in anion exchangers or channels. PMID- 9350340 TI - cDNA cloning and genomic organization of enhancer of split groucho gene from nematode Caenorhabditis elegans. AB - This first genomic Enhancer of split groucho (ESG) gene and its full length complementary DNA (cDNA) from nematode C. elegans were cloned and sequenced via homology with the corresponding Drosophila groucho cDNA. The cDNA of 2.1-Kb encodes a protein of 612 amino acids, and the nematode ESG protein is the smallest and most different in structure compared to all ESG related proteins. The gene isolated is 4,246-bp in size, including 1,219-bp promoter region. A putative TATA-box at position -1166, two consensus sequence of ACTGG, characteristic of leader binding protein-1 (LBP-1) binding motifs at position 563 and -211 and nine CAAT boxes were found in the promoter region of ESG gene. The protein-coding sequence is interrupted by five introns. The length of introns 1 to 5 is 52, 252, 87, 53 and 518 bp, respectively. The overall structural relationships of the ESG-related proteins among human, mouse, rat, Xenopus, Drosophila and nematode were also analyzed. PMID- 9350341 TI - The increased level of PDGF-A contributes to the increased proliferation induced by mechanical stimulation in osteoblastic cells. AB - Mechanical stimulation can prompt healing of bone fractures. However, it is largely unknown how osteogenesis is promoted by mechanical stimulation. In this study, we found that mechanical strain-induced proliferation of osteoblastic cells (MC3T3-E1) accompanied increased levels of platelet-derived growth factor-A (PDGF-A) mRNA, determined by quantitative reverse transcription/polymerase chain reaction. In addition, neomycin and W-7, which blocked mechanical strain-induced proliferation of the osteoblast cells, also blocked mechanical stimulation induced elevation of PDGF-A mRNA. Finally, an antibody against PDGF can inhibit physical stimulation-induced proliferation of MC3T3-E1 cells, suggesting that the increased MC3T3-E1 cells produced by mechanical stimulation at least partially depends on the increased activity of PDGF. PMID- 9350342 TI - cDNA sequence analysis of a novel neurotoxin homolog from Taiwan banded krait. AB - The cDNA encoding a novel protein was constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus (Taiwan banded krait) by reverse transcription-polymerase chain reaction. The deduced amino acid sequence of this novel protein contains 68 amino acid residues with 10 cysteine residues. Comparative sequence analyses show that it is structurally related to alpha bungarotoxin and kappa-bungarotoxins from Bungarus multicinctus venom. Eight out of the ten cysteine residues in this protein are located at the homologous positions as those in the neurotoxins. However, instead of the fifth disulfide linkage appearing in loop II of alpha-bungarotoxin and kappa-bungarotoxins, the other two cysteine residues in this novel toxin are situated at the N-terminal region. Phylogenetic analyses suggest that it probably represents a small evolutionary divergence between the long and short neurotoxins. PMID- 9350343 TI - Augmentation of protein kinase C activity and liver cell proliferation in lead nitrate-treated rats. AB - It is shown that lead alters calcium mediated cellular processes in several biological systems. Calcium enhances the activity of protein kinase C (PKC) which takes part in eliciting cell mitosis. In this study, the effects of lead nitrate on the activity of PKC enzyme were investigated in rat liver. The PKC activity was determined at 12, 24, 48, 72, 120, 168 hours after treatment with a single dose of lead nitrate in male Wistar rats. The results showed that the specific PKC activity of the purified particulate fraction was increased and reached a maximum at 24 hour, and lasted for 48 hours. This augmented activity of PKC was parallel with the increase of the lead level in the purified particulate fraction, although the protein levels of PKC alpha, PKC delta and PKC zeta were unchanged. Moreover, the frequency of mitotic cells also exhibited a significant increase, and like PKC activity, reached its maximum at 24 hour with accompany signs of liver enlargement. The results suggest that the PKC activation may be involved in promoting liver cell proliferation in lead nitrate-treated rats. PMID- 9350344 TI - Novel substrates of yeast alcohol dehydrogenase-3. 4-dimethylamino-cinnamaldehyde and chloroacetaldehyde. AB - 4-Dimethylamino-trans-cinnamaldehyde and chloroacetaldehyde are novel substrates of yeast alcohol dehydrogenase (EC 1.1.1.1). In the present work, we have reported the steady-state kinetic constants for both substrates, and their chemical reactions with the enzyme protein itself. Both substrates are potentially useful for biotechnology, chemoenzyme syntheses and analytical biochemistry. PMID- 9350345 TI - Mapping the common antigenic determinants in avidin and streptavidin. AB - An epitope scan analysis of the whole sequence of avidin and core streptavidin using polyclonal antibodies to these two antigens reveal the presence of multiple common epitopes in both the proteins. These antigenic determinants consist mostly of either identical or similar residues. The antibody recognition sites in both antigens are shown to be localized to homologous regions. PMID- 9350346 TI - Antagonistic regulation of protein kinase C-induced stimulation of fibronectin synthesis by cyclic AMP in human lung fibroblasts. AB - We studied the interaction between cAMP and protein kinase C (PKC) signaling pathways in the regulation of fibronectin synthesis in human lung fibroblasts. Phorbol myristate acetate (PMA), a PKC activator, stimulated fibronectin synthesis and its mRNA expression in both normal and transformed human lung fibroblasts (WI-38 and WI-38 VA13, respectively). On the other hand, dibutyryl cAMP (Bt2cAMP), a cAMP analogue, did not alter fibronectin synthesis in both cell lines. The combined treatment of Bt2cAMP with PMA, however, suppressed the PMA induced stimulation of fibronectin synthesis and mRNA expression in these cells. This study shows that cAMP pathway antagonizes PKC pathway in regulating fibronectin synthesis in human lung fibroblasts and provides an example of antagonistic interaction between cAMP and PKC signaling pathways. PMID- 9350347 TI - Cloning and sequencing of the secY gene homolog from Mycobacterium bovis BCG. AB - The complete nucleotide sequence of a 1,513 bp fragment of Mycobacterium bovis BCG containing the secY gene homolog and partial adk gene that encodes an adenylate kinase has been determined. The secY gene of BCG has an open reading frame of 441 amino acids with homology to the SecY protein family. Comparative analyses of the deduced amino acid sequence of additional partial ORF revealed strong similarity to the known adenylate kinases. PMID- 9350348 TI - The role of reactive oxygen species (ROS) in the effector mechanisms of human antimycobacterial immunity. AB - The aim of this study was to determine the role of reactive oxygen species (ROS) in checking the growth of intracellular mycobacteria within human phagocytes. Peripheral blood-derived neutrophils and monocyte-derived macrophages were isolated from Chronic Granulomatous Disease (CGD) patients and normal healthy human volunteers. CGD patients are known to have a defect in the NADPH oxidase pathway, resulting in their neutrophils and monocyte-derived macrophages being unable to generate oxygen radicals which are required to kill intracellular bacteria. The cells were then infected with Bacille Calmette Guerin (BCG) or Mycobacterium avium, and the bacterial growth in each cell type determined by Colony Forming Units (CFU) estimate. The results obtained indicate that there was no demonstrable inhibition in the intracellular mycobacterial growth within neutrophils or macrophages derived from either Chronic Granulomatous Disease (CGD:deficient in NADPH oxidase pathway) or normal healthy volunteers. Macrophage treatment with either IFN-gamma or TNF-alpha had no effect. PMID- 9350349 TI - Multiple cellular proteins are recognized by the adeno-associated virus Rep78 major regulatory protein and the amino-half of Rep78 is required for many of these interactions. AB - Adeno-associated virus (AAV) encoded Rep78 is a multi-functional protein which is required for AAV DNA replication, is able to regulate both AAV and heterologous gene expression at the transcriptional level, and appears necessary for site specific integration of AAV DNA into human chromosome 19. By comparison with the analogous replication protein of the polyomaviruses, large T antigen, it seemed likely that Rep78 would interact with cellular proteins to carry out at least some its functions. This study demonstrates that Rep78 is able to interact with multiple cellular proteins, from cellular extracts as measured by West(far) western, coimmunoprecipitation, and Rep78-affinity chromatography analysis. Eight cellular proteins of approximately 180, 140, 120, 95, 75, 55, 45, and 35 kDa (+/- 10%), were observed to bind Rep78 in all three assay systems. Two others, of 30 and 24 kDa, were observed in two of three assay systems. Furthermore, using truncated Rep78 proteins, it is demonstrated that the amino-terminus is required for most Rep78-cellular protein interactions. However, the extreme carboxy terminus of Rep78 was found to be all that is required for binding to the 55 kDa cellular protein. PMID- 9350350 TI - Sucrose-phosphate synthase in tree species: light/dark regulation involves a component of protein turnover in Prosopis juliflora (SW DC). AB - Light dependent modulation of sucrose-phosphate synthase activity (SPS; EC 2.4.1.14) was studied in a tree species, namely Prosopis juliflora. In this paper we demonstrate that cycloheximide, an inhibitor of cytoplasmic protein synthesis, when fed to detached leaves of P. juliflora through transpiration stream in the dark or in light completely prevents in vivo light activation of Vlim and Vmax activities of SPS. In case of spinach, however, cycloheximide feeding affects only Vlim activity while Vmax activity remained unchanged. In contrast, chloramphenicol, an inhibitor of protein synthesis in chloroplast has no effect on the light activation of SPS in Prosopis. The treatment with cycloheximide showed slight reduction in the rate of O2 evolution indicating that cycloheximide had very little effect on overall photosynthesis. These results indicate that short term protein turnover of the SPS protein and some other essential component(s) (e.g., a putative protein that modifies SPS activity) is one of the primary steps in a complex and unique regulatory cascade effecting the reversible light activation of SPS. PMID- 9350351 TI - Characterization of the orf31-petG gene cluster from the plastid genome of Populus deltoides. AB - The orf31-petG gene cluster is located approximately 1.2 kb away from the psbEFLJ operon in the chloroplast genome of Populus deltoides. The orf31 (ycf7) encodes an unidentified polypeptide while the petG gene encodes subunit V of an important component, cytochrome b6/f complex, involved in photosynthetic electron transport. We have determined the nucleotide sequence of the orf31-petG gene cluster from the plastid genome of a tree, Populus deltoides. Our sequence analysis suggests that these genes possess high homology with the published sequences of these genes from other plants. Northern analysis suggests development dependent transcription of the orf31-petG cluster in leaves. PMID- 9350352 TI - Fucoidan inhibits leukocyte recruitment in a model peritoneal inflammation in rat and blocks interaction of P-selectin with its carbohydrate ligand. AB - Neutrophil recruitment into systemic inflammatory sites in vivo is thought to be initiated by selectin-mediated endothelial adherence. The effect of fucoidan (natural sulfated polymer of L-fucose) on the selectin dependent PMN migration into rat peritoneum following the induction of inflammation by peptone injection was studied. Peritonitis was characterized by an increase in the total cell number (from 45.3 x 10(6) to 91.6 x 10(6)/rat), and by highly elevated PMN content (from 0.2% to 58%) in the rat peritoneal cavity 3 h after peptone injection. Intravenous administration of fucoidan was found to reduce, in a dose dependent manner, neutrophil migration into peritoneum. Fucoidan in a dose as low as 0.8 mg per rat caused 96.8% reduction of neutrophil extravasation. The inhibitory effect of fucoidan was also dependent on the time intervals between the peptone and fucoidan injections. The maximal inhibitory effect of fucoidan was observed within the first 15 min after the induction of peritonitis and it was maintained at a level of 80% during 1.5 h. Administration of fucoidan 2.5 h after peptone injection had practically no effect on PMN extravasation. Since P selectin is known to play a key role at the earlier stages of PMN extravasation, it was suggested that the inhibitory effect of fucoidan was mostly due to its interaction with P-selectin. The in vitro experiments demonstrated the high affinity of fucoidan for both isolated P-selectin and P-selectin in plasma membranes of activated platelets. PMID- 9350353 TI - Evidence for the essential histidine residues in geranylgeranyl transferase type I from bovine testis. AB - Geranylgeranyl transferase was purified 30,000-fold by sequential use of 30-50% ammonium sulfate fractionation, Q-Sepharose anion exchange chromatography, Phenyl Superose hydrophobic interaction chromatography, Sephacryl S-300 gel filtration chromatography, and peptide (YREKKFFCAIL) affinity chromatography. Geranylgeranyl transferase, when incubated with diethyl pyrocarbonate (DEPC), a histidine specific reagent, shows time-dependent inactivation, and the activity is restored by the addition of neutral hydroxylamine. The inactivation follows pseudo-first order kinetics with a second order rate constant of 0.319 M-1min-1. The overall results thus provide evidence that a histidine residue in the active site is involved in the catalytic mechanism of the geranylgeranyl transferase reaction. PMID- 9350354 TI - Biomolecular computer: roots and promises. AB - There are two basic issues inherent in contemporary molecular and biomolecular computing and urgent for its future development. They are: (i) fundamentals, that is general principles, starting points and correlation's between different approaches to design biomolecular information processing devices, (ii) working criteria, that is goals and principles for the choice of the device designing characteristics to achieve these goals. These issues are common to all major approaches of contemporary biomolecular computing. Increasing behavioral complexity of an information processing device seems to be a criterion for designing biomolecular means capable of solving problems of high computational complexity. PMID- 9350355 TI - Isomorphism between cell and human languages: molecular biological, bioinformatic and linguistic implications. AB - The concept of cell language has been defined in molecular terms. The molecule based cell language is shown to be isomorphic with the sound- and visual signal based human language with respect to ten out of the 13 design features of human language characterized by Hockett. Biocybernetics, a general molecular theory of living systems developed over the past two and a half decades, is found to provide a physical theory underlying the phenomenon of cell language. The concept of cell language integrates bioenergetics and bioinformatics on the one hand and reductionistic and holistic experimental data on the other to account for living processes on the molecular level. The isomorphism between cell and human languages suggests that the DNA of higher eucaryotes contains two classes of genes--structural genes corresponding to the lexicon and 'spatiotemporal genes' corresponding to the grammar of cell language. The former is located in coding regions of DNA and the latter is predicted to reside primarily in noncoding regions. The grammar of cell language is identified with the mapping of the nucleotide sequences of DNA onto its 4-dimensional folding patterns that control the spatiotemporal evolution of gene expression. Such a mapping has been referred to as the second genetic code, in contrast to the first genetic code which maps nucleotide triplets onto amino acids. The cell language theory introduces into biology the linguistic principle of 'rule-governed creativity,' leading to the formulation of the concept of 'rule-governed creative molecules' or 'creations.' This concept sheds new light on molecular biology, bioinformatics, protein folding, and developmental biology. In addition, the cell language theory suggests that human language is ultimately founded on cell language. PMID- 9350356 TI - Models of CO2 concentrating mechanisms in microalgae taking into account cell and chloroplast structure. AB - Detailed mathematical models have been developed for the functioning of CO2 concentration mechanisms in microalgae. The models treat a microalgal cell as several compartments: pyrenoid, chloroplast stroma, cytoplasm and periplasmic space. Cases for both the active bicarbonate transport through the plasmalemma and the passive CO2 diffusion through it with the subsequent concentrating of CO2 inside the chloroplast are analyzed. CO2 evolution from bicarbonate inside the pyrenoid is modeled. The great diffusion resistance for CO2 flux from the pyrenoid is caused by a starch envelope and the concentric thylakoid membranes surrounding it. The role of carbonic anhydrase in the periplasmic space, cytoplasm and inside the chloroplast is evaluated numerically. The models also offer an explanation for the absence of 'short-circuited' inorganic carbon fluxes between the external medium and the cytoplasm under active bicarbonate transport through the plasmalemma and in the presence of carbonic anhydrase in the cytoplasm. If the cytoplasm is driven from the space between a chloroplast envelope and plasmalemma upon the microalgae adaptation to low concentration of the dissolved inorganic carbon, the inorganic carbon leak might be avoided. The models reproduce accurately the majority of known experimental data. The high efficiency of CO2 concentrating mechanisms in microalgae can be explained by a considerable diffusion resistance for CO2 flux from the pyrenoid and by the effective scavenging of CO2 leaking outward from the chloroplast to cytoplasm and from cell to periplasmic space. PMID- 9350357 TI - Diffusion model for growth factors--cell receptors interaction. AB - A theoretical model describing the interaction between growth factor molecules and their receptors from the cell surface within an 'off-centre' diffusion approximation is developed in this paper. It is assumed that a number of non interacting particles (growth-factor molecules) more diffusively in the presence of traps (specific receptors) which are located at the surfaces of a number of cells uniformly distributed in space. The diffusion equations system is solved by a perturbative method. The model predicts a nonlinear dependence of the receptor occupation and of the time needed to reach a threshold value of receptor occupation on the total number of cell receptors. An autocrine binding, when the ligand is secreted by the responsive cells accounts for the interaction between IL-2 and helper T-cells. PMID- 9350359 TI - The cost of cancelling surgery. PMID- 9350358 TI - Protein stability; optimization of electrostatic contributions by partially neutralizing surface ionic charges. AB - 'Partial Charge-Neutralization Method' is developed to study influence of the relative amount of positive and negative charges in proteins on their structural stability. A given number of either positively or negatively charged groups are neutralized in all of their possible combinations to generate a whole set of distinct species. The Coulomb energy of each species is calculated by numerically solving the Poisson-Boltzmann equation for aqueous solutions. Partial neutralization of lysine residues of tuna cytochrome c in aqueous solution at neutral pH with the Debye-Huckel screening parameter kappa = 1 nm-1 reproduces qualitatively well the destabilization of acetylated cytochrome c observed in physicochemical measurements at pH 7. The stabilization of its molten globule state at pH 2 is also studied with the present method. It is shown that the electrostatic contribution to the structural stability of natural proteins can be optimized by changing the difference in number of their positive and negative charges. PMID- 9350360 TI - A "zero tolerance for overtime" increases surgical per case costs. AB - PURPOSE: One technique which some hospitals have used in an attempt to control Operating Room costs is a "zero tolerance for overtime" policy. We used a case cost analysis to determine if this policy was always cost effective. METHOD: A case cost analysis was designed based on a "test case" which would start late in the day. The case would last for three hours of which 1 1/2 hr would be during regular hours, and 1 1/2 hr would incur overtime. Costs were analysed using a "patient pays," "society pays," and "hospital pays" analysis. Costs were based on figures determined from the SMBD-Jewish General Hospital budget, Quebec Health Insurance fees, and Government of Canada statistics. RESULTS: Regardless of who pays, in this case scenario it was more cost effective to proceed than to postpone surgery. Costs of proceeding with the surgery in the "patient pays," "society pays," and "hospital pays" models were $1,832.00, $1,227.40, and $1,215.00 respectively. The costs of postponing the surgery in the same three models were $1,937.00, $1,336.80, and $1,436.00. CONCLUSION: A "zero tolerance for overtime" policy may be too rigid to be consistently cost effective. PMID- 9350361 TI - Continuous spinal anaesthesia using a standard epidural set for extracorporeal shockwave lithotripsy. AB - PURPOSE: Continuous spinal anaesthesia (CSA) offers considerable advantages over "single shot" spinal or epidural anaesthesia since it allows titration of anaesthesia using small doses of local anaesthetics (LA). We evaluated the feasibility of CSA using a standard epidural set for extracorporeal shockwave lithotripsy (ESWL). METHODS: Charts of 100 consecutive CSAs for ESWL were retrospectively reviewed. Lumbar CSA was performed using a 20G epidural catheter through an 18G Tuohy needle. The CSA was preplanned, or followed inadvertent dural puncture. Small LA boluses were injected to achieve the desired sensory level of anaesthesia. Demographic data, anaesthetic duration, LA doses, the most cephalad sensory level to pinprick, arterial blood pressure, heart rate, use of systemic sympathomimetics and complications were recorded. RESULTS: Mean age was 66.2 +/- 9.9 (SD). The ASA status was III-IV in 54.1% and 5.5% of the preplanned and inadvertent patients, respectively. In 85 anaesthetics, hyperbaric bupivacaine 0.1% (9.7 +/- 7.5 mg) was used as the sole anaesthetic. Sensory level was T4-T8. Maximal decrease in systolic and diastolic blood pressures and heart rate was 19.0 +/- 9.8%, 13.4 +/- 13.3%, and 7.2 +/- 11.7 respectively. Intravenous sympathomimetics were used in nine of 82 (11.0%) preplanned, and in six of 18 (33.3%) inadvertent anaesthetics. Post dural puncture headache appeared following two of 82 (2.5%) preplanned, and four of 18 (22.2%) inadvertent anaesthetics. No postanaesthetic neurological deficit was detected. CONCLUSION: Continuous spinal anaesthesia, using a standard epidural set and hyperbaric bupivacaine is feasible for ESWL in high risk patients. Inadvertent dural puncture does not preclude CSA under these circumstances. PMID- 9350362 TI - Enhanced pain management for postgastrectomy patients with combined epidural morphine and fentanyl. AB - PURPOSE: To determine whether clinical advantages could be demonstrated by epidural fentanyl given in addition to epidural morphine for postgastrectomy analgesia. METHODS: One-hundred and twenty two patients undergoing elective gastrectomy were prospectively studied in a randomised, double-blind fashion. All patients received epidural lidocaine 1.5% with epinephrine (1:200,000) followed by light general anaesthesia for surgical anaesthesia. They were assigned to four groups according to the combinations of each epidural opioid: 2 mg morphine alone, 2 mg morphine + 100 micrograms fentanyl, 4 mg morphine alone, and 4 mg morphine + 100 micrograms fentanyl. Morphine and fentanyl were given epidurally approximately 60 and 15 min, respectively, before the completion of surgery. RESULTS: Addition of epidural fentanyl to both doses of morphine not only decreased intensity of pain associated with coughing during the early postoperative period, but also prolonged the time until the first analgesic request at each morphine dose studied. Of the combination doses, 4 mg morphine + 100 micrograms fentanyl provided the longest time to the first request for analgesic, and was associated with least amount of postoperative analgesic supplement and best patient satisfaction without increasing incidence of side effects. CONCLUSION: The addition of 100 micrograms fentanyl to 2 mg or 4 mg epidural morphine provides clinical advantages over morphine alone for post gastrectomy analgesia. PMID- 9350363 TI - Trial of three methods of intraoperative bupivacaine analgesia for pain after paediatric groin surgery. AB - PURPOSE: To evaluate the relative effectiveness of three techniques of regional anaesthesia in the provision of postoperative analgesia in children. METHODS: Random assignment was made of 183 children scheduled for groin surgery to one of three groups. Bupivacaine 0.5% plain (2 mg.kg-1) was injected by the surgeon after skin incision. Group A received wound infiltration. Group B had regional nerve blockade. Group C had a combination of both methods. Postoperatively, pain was assessed using the CHEOPS behavioural scale at half hourly intervals until discharge home. Satisfactory pain control was arbitrarily defined as a CHEOPS score of < or = six. Potential differences among the groups were sought using graphical representation of mean pain scores, the frequencies of maximum pain scores, and the incidence of postoperative vomiting and oral analgesic consumption. RESULTS: Fifteen patients had to be excluded from analysis. This left 61 patients in Group A, 55 in Group B, and 52 in Group C. There were no demographic differences among the groups. No differences were demonstrated among the groups either in CHEOPS pain scores at any observation point (P = > 0.8), or in the incidence of vomiting or need for postoperative analgesia. (P = 0.52 and P = 0.41 respectively). Overall, 80% of the observations made (1,135/1,425) met our definition of satisfactory pain control. A post hoc calculation of the power of the study confirmed sufficient power to detect a 5% difference among groups. CONCLUSION: All three methods achieved analgesia with 80% of the pain scores meeting our definition of satisfactory pain control. None of the techniques enjoyed any apparent advantage. PMID- 9350364 TI - Treatment of intrathecal morphine-induced pruritus following caesarean section. AB - PURPOSE: To compare both the efficacy and cost of nalbuphine and diphenhydramine in the treatment of intrathecal morphine-induced pruritus following Caesarean section. METHODS: Eighty patients, undergoing elective Caesarean section under spinal anaesthesia, were randomized, in a prospective, double-blind trial, to receive either nalbuphine (Group NAL) or diphenhydramine (Group DIP) for the treatment of SAB morphine-induced pruritus. All patients received an intrathecal injection of 10-12 mg hyperbaric bupivacaine 0.75% and 200 micrograms preservative free morphine. Postoperative pruritus was assessed, using a visual analogue scale (VAS), for 24 hr. Pruritus treatment was administered upon patient request and by a nurse blinded to the treatment given. Patients who failed to respond to three doses of the study drug were deemed treatment failures. Patient satisfaction was assessed with a questionnaire given 24 to 48 hr after surgery. Direct drug costs were calculated based on the pharmacy provision costs as of April 1996. RESULTS: Eighty patients were enrolled and 45 requested treatment for pruritus. Patients treated with NAL (n = 24) were more likely to achieve a VAS score of zero with treatment (83% vs 43%, P < 0.01), had a higher delta VAS following treatment (4 +/- 2 vs 2 +/- 2, P < 0.003), and experienced fewer treatment failures (4% vs 29%, P < 0.04), than those treated with DIP (n = 21). Group NAL patients were also more likely to rate their pruritus treatment as being good to excellent (96% vs 57%, P < 0.004). Direct drug costs were higher for NAL than for DIP ($6.4 +/- 3.1 vs $1.7 +/- 0.7, respectively, P < 0.0001). CONCLUSION: Nalbuphine is more effective than diphenhydramine in relieving pruritus caused by intrathecal morphine and the cost differences are small. PMID- 9350365 TI - Haemodynamic and catecholamine changes during rapid sevoflurane induction with tidal volume breathing. AB - PURPOSE: To compare haemodynamic and plasma catecholamine changes with rapid (three minute) inhalational anaesthesia induction with tidal volume breathing of sevoflurane 7%, conventional (seven minute) slow inhalation induction with increasing sevoflurane concentration up to 5%, and induction with thiamylal i.v. METHODS: Twenty-four patients were randomly divided into three groups of eight. In Group A, anaesthesia was induced with tidal volume breathing of sevoflurane 7% (inspiratory concentration) and nitrous oxide 50% in oxygen (total flow; 6 l.min 1) for three minutes: Group B received conventional slow induction for seven minutes and increasing sevoflurane concentration by 0.5% every two or three breaths up to 5% with nitrous oxide 50% in oxygen: and Group C received 5 mg.kg-1 thiamylal with an inhalation of 100% oxygen. Blood pressure, heart rate, rate pressure product, and plasma concentrations of epinephrine and norepinephrine were measured. RESULTS: There were no changes in blood pressure before or after intubation in Group A (sevoflurane 7%) whereas both were increased in patients in Group C (thiamylal). Changes in heart rate and rate pressure product were not different for the two inhalation groups. Plasma epinephrine concentrations decreased in all the three groups. Norepinephrine concentrations were increased before intubation in both inhalation groups but not in the thiamylal group. CONCLUSION: Rapid induction of anaesthesia with sevoflurane 7% and tidal volume breathing for three minutes induced less haemodynamic changes than the other methods studied and has no inhibitory effect on sympathetic activity. PMID- 9350366 TI - Interactions between nicardipine and enflurane, isoflurane, and sevoflurane. AB - PURPOSE: During nicardipine induced hypotension, different inhalational anaesthetics may have different effects on haemodynamic variables, sympathetic function and drug metabolism. Therefore, the haemodynamic effects and pharmacokinetics of nicardipine were studied in the presence of the three inhalation anaesthetics enflurane, isoflurane and sevoflurane. METHODS: Thirty patients scheduled for neurosurgery were randomly assigned to one of three anaesthetic techniques: enflurane, isoflurane or sevoflurane. Nicardipine (0.017 mg.kg-1) was administered during stable anaesthesia and the following measurements made for 30 min: blood pressure, heart rate, and plasma concentration of norepinephrine, epinephrine and nicardipine. RESULTS: With sevoflurane, plasma concentrations of nicardipine, five minutes after administration, (39.8 +/- 3.5 ng.ml-1, mean +/- SEM) were higher (P < 0.05) than in the other two groups (28.3 +/- 2.9 ng.ml-1, 32.6 +/- 4.3 ng.ml-1, enflurane and isoflurane, respectively). With isoflurane, the approximated half-life of nicardipine (14 +/- 4 min) was shorter and clearance (2.1 +/- 0.3 l.min-1) more rapid. Peak heart rates were similar in all groups but elevated rates continued longer with isoflurane (> 30 min). Nicardipine-induced reduction in blood pressure was greater with sevoflurane but low pressures persisted for longer with isoflurane. Plasma catecholamine concentrations increased with isoflurane and enflurane, but not with sevoflurane: considerably higher epinephrine concentrations were seen with isoflurane. CONCLUSION: This study showed that the action of nicardipine is modified by different inhalational anaesthetic agents. Nicardipine has a prolonged duration of action in the presence of isoflurane and produces greater initial hypotension with sevoflurane. PMID- 9350367 TI - Comparison of cerebral oximeter and evoked potential monitoring in carotid endarterectomy. AB - PURPOSE: To assess the cerebral oximeter, which measures regional oxygen saturation (rSO2) continuously and noninvasively, as a cerebral monitor during carotid endarterectomy (CEA). The rSO2 was compared with Somatosensory Evoked Potentials (SSEPs) as an indicator for shunting and as a predictor of postoperative neurological deficits. METHODS: Seventy-two consenting patients undergoing CEA with general anaesthesia were studied. Normocarbia, normothermia and normotension were maintained. Cerebral monitoring consisted of bilateral median nerve SSEPs and the INVOS 3100 cerebral oximeter with the sensor pad placed on the ipsilateral forehead. Decreases in SSEP amplitude of 50% and in rSO2 of 10% were considered clinically significant. Neurological assessment was performed at emergence from anaesthesia, 24 hr postoperatively and at discharge. The rSO2 changes were compared with SSEP changes and with neurological deficits. Statistical analysis was with chi square and analysis of variance P < 0.05 was considered significant. RESULTS: During carotid artery clamping, rSO2 decreased from 72 +/- 8% to 68 +/- 9% and mean arterial blood pressure increased from 92 +/ 14 mmHg to 98 +/- 14 mmHg. In four patients, the carotid artery was shunted because of SSEP changes after cross-clamping. Five patients had > or = 10% decreases in rSO2 following clamp application. Changes in both SSEP and rSO2 occurred in two patients. Three of the four shunted patients had transient postoperative neurological deficits. One patients had a transient deficit without changes in either monitor. There were no persistent postoperative deficits. Compared with SSEPs, rSO2 had a sensitivity of 50% and a specificity of 96%. CONCLUSION: Clinical experience with this evolving technology is ongoing. Its role in neurovascular procedures has yet to be established. PMID- 9350368 TI - Cardiovascular responses to tracheal extubation or LMA removal in normotensive and hypertensive patients. AB - PURPOSE: This study was undertaken to evaluate the haemodynamic changes of tracheal extubation or removal of a laryngeal mask airway (LMA) in normotensive and hypertensive patients. METHODS: In a randomized trial of normotensive and hypertensive patients (n = 40 of each), tracheal extubation or LMA removal was performed. Changes in heart rate (HR), mean arterial pressure (MAP) and rate pressure product (RPP) were measured before and 1, 2, 3, 5, and 10 min after tracheal extubation or LMA removal. RESULTS: In normotensive patients, HR, MAP and RPP increased following tracheal extubation or LMA removal, and remained elevated for a maximum three minutes (P < 0.05). In hypertensive patients, the haemodynamic increases in response to extubation or LMA removal were observed for up to five minutes (P < 0.05). The immediate cardiovascular responses to extubation were greater than those related to LMA removal in both normotensive and hypertensive patients (normotensive: HR; 95 +/- 14 vs 81 +/- 11, MAP; 124 +/- 18 vs 106 +/- 10, RPP; 14,951 +/- 2720 vs 10,654 +/- 1898, hypertensive: HR 105 +/- 10 vs 87 +/- 13, MAP; 146 +/- 17 vs 119 +/- 12, RPP; 20,492 +/- 1674 vs 12,862 +/- 2115, mean +/- SD, P < 0.05). Following extubation or LMA removal, these haemodynamic variables increased more markedly in hypertensive patients than in normotensive patients (P < 0.05). CONCLUSION: Removal of LMA is associated with less cardiovascular change than tracheal extubation in both normotensive and hypertensive patients. PMID- 9350369 TI - Same day drainage and removal of a giant ovarian cyst. AB - PURPOSE: An unusual case of a giant ovarian cyst was successfully anaesthetized with a combination of epidural followed by general anaesthesia. The method was chosen to avoid circulatory depression and re-expansion pulmonary oedema in removal of a giant tumour in a woman who did not understand the nature of her disease. CLINICAL FEATURES: A 58-yr-old woman (107.6 kg, 150 cm and abdominal girth: 163.5 cm) was admitted for removal of a giant ovarian cyst. There was gross-pitting oedema of both legs and an elevated diaphragm but no pleural effusion. She did not understand the severity of her disease. It was decided to drain the cyst gradually, followed by total surgical removal on the same day. An epidural catheter was inserted at the L3-4 interspace with the patient in the left lateral position and, under epidural anaesthesia, 44.3 L fluid were drained over two hours without producing circulatory depression or pulmonary oedema. General anaesthesia was induced, with the patient in the supine position, by slow injection of 10 mg midazolam, 100 micrograms fentanyl and inhalation of nitrous oxide 50% in oxygen, and maintained with adding epidural block using lidocaine 1.5% and bupivacaine 0.5%, and sevoflurane 0.4 to 0.8%. During surgery, the volume of infused fluid was carefully controlled with central venous pressure monitoring. Ulinastatin, a protease inhibitor, was infused to prevent pulmonary oedema. No circulatory depression nor pulmonary oedema occurred perioperatively. CONCLUSION: For the removal of a giant ovarian cyst, slow drainage over two hours under epidural anaesthesia may safely precede later removal of the cyst on the same day under general anaesthesia. PMID- 9350370 TI - Anaesthetic management of a patient with Leigh's syndrome. AB - PURPOSE: Leigh's syndrome, a progressive neurodegenerative disorder of infancy and childhood, is clinically characterized mainly by developmental delay, nervous system dysfunction and respiratory abnormalities such as aspiration, wheezing, breathing difficulties, gasping, hypoventilation and apnoea. Acute exacerbation and respiratory failure may follow surgery, general anaesthesia or intercurrent illnesses. Hyperlecithinemia is variably present. Histopathological findings include necrosis, vascular proliferation, astrocytosis and demyelination of several brain areas. We present a 30-month-old patient with Leigh's syndrome anaesthetized for extracorporeal shockwave lithotripsy, and describe the anaesthetic considerations. CLINICAL FEATURES: Leigh's syndrome was diagnosed at five months of age based on failure to thrive, lethargy, hypotonicity, choreo athetosis and lactic acidaemia, with basal ganglia hypodense areas demonstrated by brain computerized tomographic scan. Muscle pyruvate dehydrogenase complex and NADH coenzyme Q oxidoreductase activity were 25% and 13% of control. No preoperative respiratory symptoms or signs were present. Preoperative fasting lasted two hours and gastric aspiration was negative. Anaesthesia was induced with ketamine and midazolam im, and N2O in oxygen, and maintained with propotol and N2O. No volatile anaesthetics were used. Intravenous fluids given were 1/2 normal saline and glucose 5% administered. Besides laryngospasm during anaesthetic induction, relieved by sublingual succinylcholine injection, the perianaesthetic course was uneventful. The lungs were mechanically ventilated and lithotripsy was performed. No adverse sequelae have occurred, and the patient was discharged one day later. CONCLUSION: Perioperative management of patients with Leigh's syndrome requires cautious attention to the metabolic, neurological and respiratory aspects of the disease, and appropriate selection of anaesthetic drugs. PMID- 9350371 TI - Cerebral protection using retrograde cerebral perfusion during hypothermic circulatory arrest. AB - BACKGROUND: Retrograde cerebral perfusion through the superior vena cava (SVC) has been proposed to protect the brain from ischaemic injury during profound hypothermic circulatory arrest (PHCA). Its contribution to cerebral protection is unclear. Furthermore, the addition of anaesthetic or vasodilating agents to the SVC perfusate to enhance brain protection, has never been described. METHODS: In three patients undergoing repair of the ascending aorta utilizing PHCA, the upper body was retrogradely perfused with cold (16 degrees C) blood through the SVC by the cardiopulmonary bypass pump. Electroencephalographic activity was monitored using a computerized electroencephalographic monitor (Cerebro Trac 2500, SRD). Perfusion pressure was measured at a port in the cannula connector. Etomidate or thiopentone was injected into the SVC perfusate to arrest reappearing electroencephalographic activity. Nitroglycerin or nitroprusside was injected into the perfusate to increase retrograde flow and maintain a constant perfusion pressure. RESULTS: During PHCA periods of up to 61 min, recurrent electroencephalographic activity was abolished by the retrograde administration of small boluses of etomidate (total 50 mg) or thiopentone (total 500 mg). Nitroprusside (100 micrograms) and nitroglycerin (2 micrograms.kg-1.min-1) increased retrograde flow from 220 to 550 and 660 ml.min-1, respectively, while maintaining perfusion pressure (25-26 mmHg). Recovery from anaesthesia and surgery was uneventful, with no adverse neurological sequelae. CONCLUSION: Injection of anaesthetic agents into the retrograde SVC perfusate during PHCA, can suppress reoccurring electroencephalographic activity and retrograde injection of vasodilators can facilitate an increase in perfusion. It is suggested that both may augment brain protection. PMID- 9350372 TI - Venous gas embolism--a comparison of carbon dioxide and helium in pigs. AB - PURPOSE: The use of helium for insufflation during laparoscopic surgery avoids hypercarbia and acidosis associated with absorbed CO2, but the effects of helium gas embolism are unknown. We compared the effects of CO2 with He gas embolism on survival, haemodynamic variables, oxygenation, and ventilation in pigs. METHODS: Anaesthetized juvenile pigs were given progressively larger boluses of either CO2 (n = 5) or He (n = 4) into the right atrium. Measurements of haemodynamic variables, oxygenation, and PETCO2 were made before and after each gas injection. RESULTS: All animals survived injection of 300 ml CO2 while no animal survived more than 120 ml He (P < 0.01). Mean arterial pressure decreased more after 60 ml He (99 +/- 14 to 44 +/- 20 mmHg) than after 60 ml CO2 (110 +/- 12 to 88 +/- 14 mmHg, P < 0.001). Cardiac output did not change at any injection volume. The PETCO2 decreased more after 60 ml He (30 +/- 2 to 3 +/- 6 mmHg) than after 60 ml CO2 (35 +/- 3 to 30 +/- 3 mmHg, P < 0.001). Only the He group showed a decrease in PaO2 (190 +/- 51 to 68 +/- 22 mmHg at 60 ml, P < 0.05). CONCLUSION: Helium gas embolism has a greater deleterious effect than CO2 gas embolism on survival, MAP, PETCO2, and PaO2. These different effects of gas embolism should be recognized when considering the use of helium or other insoluble gases for abdominal laparoscopic insufflation. PMID- 9350373 TI - Isoflurane inhibits endothelium-mediated nitric oxide relaxing pathways in the isolated perfused rabbit lung. AB - PURPOSE: The role of volatile anaesthetics on nitric oxide (NO)-dependent relaxation is unclear in the pulmonary circulation. We examined the effects of isoflurane on NO-dependent relaxation in isolated perfused rabbit lungs. METHODS: Eighteen rabbit lungs were perfused in a constant-flow recirculation manner. In study 1 (n = 12), acetylcholine (ACh, 4 x 10(-10)-10(-8) M) or nitroglycerine (NTG, 6 x 10(-10)-10(-8) M) was cumulatively injected into the pulmonary artery in the absence or presence of isoflurane (1, 2 MAC). In study 2 (n = 6), ACh was injected as in study 1 in the presence or absence of N omega-nitro-L-arginine methyl ester (L-NAME, 100 microM), an NO synthesis blocker. In all experiments, indomethacin was administered to prevent formation of vasoactive prostanoid metabolites, and the pulmonary vessels were preconstricted with prostaglandin F2 alpha (PGF2 alpha) infused before ACh or NTG injection. The ACh- or NTG-induced relaxation was expressed as % decrease in PGF2 alpha preconstriction. RESULTS: Isoflurane at 2 MAC attenuated the dose-dependent relaxation to ACh at doses of 4 x 10(-9) M and 4 x 10(-8) M from 27.8 +/- 4.3% and 38.8 +/- 5.3% to 17.0 +/- 3.5% and 25.5 +/- 4.9%, respectively (P < 0.05). Isoflurane did not change the dose dependent relaxation to NTG and L-NAME abolished the ACh-induced relaxation. CONCLUSION: Isoflurane inhibited NO-dependent relaxation in the pulmonary circulation at a site distal to the endothelial cell receptor-mediated responses but proximal to guanylate cyclase activation of vascular smooth muscle. Acetylcholine-induced relaxation in isolated perfused rabbit lungs was regulated primarily by NO. PMID- 9350374 TI - Midazolam reverses histamine-induced bronchoconstriction in dogs. AB - PURPOSE: Midazolam has been used clinically as a sedative and as an anaesthetic induction agent. However, the bronchodilating effects of midazolam have not been comprehensively evaluated. We sought to determine relaxant effects of midazolam on the airway. METHODS: After our Animal Care Committee approved the study, eight mongrel dogs were anaesthetized with 30 mg.kg-1 pentobarbitone iv, and were paralysed with 200 micrograms.kg-1.hr-1 pancuronium. The trachea was intubated with an endotracheal tube (ID 7 mm) that had a second lumen for insertion of a superfine fibreoptic bronchoscope (OD 2.2 mm) to measure the bronchial cross sectional area (BCA) continuously. The tip of the bronchoscope was placed at the level of the second or third bronchial bifurcation of the right bronchus. A videoprinter printed the BCA which was then measured with a NIH image program. Bronchoconstriction was produced with histamine (H) 10 micrograms.kg-1 followed by 500 micrograms.kg-1.hr-1. Thirty minutes later, 0 [saline], 0.01, 0.1 and 1.0 mg.kg-1 midazolam and 25 micrograms.kg-1 flumazenil were given. The BCA was assessed before (basal area) and 30 min after the start of H infusion, and was also measured five minutes after each midazolam and flumazenil iv. At the same time, arterial blood was sampled for plasma catecholamine measurement. RESULTS: Histamine infusion decreased BCA to 49.7 +/- 17.3% of basal BCA. More than 0.1 mg.kg-1 midazolam increased BCA up to 71.7 +/- 15.3% of the basal (1.0 mg.kg-1) (P < 0.01). Plasma adrenaline concentration was decreased from 6.9 +/- 3.8 to 3.7 +/- 1.9 ng.ml-1 by 1.0 mg.kg-1 midazolam (P < 0.05). Flumazenil did not antagonize the relaxant effect of midazolam but reversed the inhibitory effect of midazolam on histamine-induced adrenaline release. CONCLUSION: Midazolam has a spasmolytic effect on constricted airways but this bronchodilatation was not reversed by flumazenil. PMID- 9350375 TI - Factors influencing MAC reduction after cardiopulmonary bypass in dogs. AB - BACKGROUND: Anaesthetic requirements may be reduced following surgery employing cardiopulmonary bypass (CPB). This study, in dogs, determined the role of a) volatile agents (enflurane [E] vs isoflurane [I]), b) oxygenator (bubble [B] vs membrane [M]), and c) presence [FL] vs absence [NoFL] of an in-line arterial filter in the bypass circuit in altering anaesthetic requirements following CPB. METHODS: Male mongrel dogs were anaesthetized with either enflurane (n = 24) or isoflurane (n = 24). They were randomly assigned to one of eight groups (n = 6 per group); Group 1 (E/B/FL), Group 2 (E/M/FL), Group 3 (E/M/NoFL), Group 4 (E/B/NoFL), Group 5 (I/M/FL), Group 6 (I/B/FL), Group 7 (I/M/NoFL) or Group 8 (I/B/NoFL). MAC was determined using the tail-clamp method at hourly intervals, twice before and three times after a one hour normothermic perfusion using aortoatrial cannulation and CPB. RESULTS: Prior to CPB, MAC was reproducible (enflurane: MAC1 2.17 +/- 0.29 vs MAC2 2.14 +/- 0.28%; isoflurane: MAC1 1.42 +/- 0.31 vs MAC2 1.41 +/- 0.33%) and differed among groups only for the volatile agent employed. Following CPB, MAC was reduced in all groups (P < 0.05 vs pre-CPB measurements) except Group 1 (E/B/FL). The degree of MAC reduction in other groups ranged from 39-64% and was not different based on type of agent employed, use of a membrane or bubble oxygenator, or presence or absence of an in-line arterial filter. CONCLUSION: In dogs, MAC reduction following CPB was variable, not related to type of volatile agent employed, use of a membrane or bubble oxygenator, or presence or absence of an in-line arterial filter. The explanation for reductions in anaesthetic requirements following CPB in this model remains speculative. PMID- 9350376 TI - Postoperative rocuronium reparalysis. PMID- 9350377 TI - Masquerading neuropathies. PMID- 9350378 TI - Detecting endobronchial intubation during breast surgery. PMID- 9350379 TI - Enlarged parathyroid gland--airway problem. PMID- 9350380 TI - Abbreviated laparotomy and planned re-operation. PMID- 9350381 TI - Convergence of occipital nerve and superior sagittal sinus input in the cervical spinal cord of the cat. AB - Co-existence of facial and occipital pain may occur in occipital neuralgia, migraine and cluster headache; suggesting convergence of trigeminal and cervical afferents. Such convergence has been shown in humans and other animals, but the site and extent of this are uncertain. In anaesthetized adult cats, the superior sagittal sinus and occipital nerve were stimulated electrically, and extracellular recordings made in the dorsolateral area of the upper cervical cord using glass-coated tungsten electrodes. Of 49 units in 10 cats, 33 (67%) had input from the superior sagittal sinus and the occipital nerve. Thirteen (27%) had superior sagittal sinus input and 3 (6%) had occipital nerve input. Convergent receptive fields were identified mechanically in 7 units. These experiments in cats show convergent input from occipital nerve and superior sagittal sinus on dorsolateral area units in two-thirds of cases studied. This experimental site of trigeminocervical convergence may relate to referral of pain in occipital neuralgia and other headaches. PMID- 9350382 TI - Migraine therapy: relationship between serotonergic contractile receptors in canine and rabbit saphenous veins to human cerebral and coronary arteries. AB - Canine and rabbit vascular contractile responses to serotonergic agonists have been used to predict antimigraine efficacy for several antimigraine agents, including sumatriptan. The purpose of the present study was to establish the assumed predictive value of contractile responses in canine and rabbit saphenous veins to contractile efficacy for a series of agonists in human cerebral and coronary arteries and to understand better the receptors mediating such responses. The canine and rabbit saphenous veins contracted similarly (both qualitatively and quantitatively) to a series of structurally diverse serotonergic agonists, suggesting that the receptors mediating serotonin-induced contractility in these tissues were similar. In addition, the contractile potency (estimated as EC50 values) for these structurally diverse serotonergic agonists in either the rabbit or canine saphenous vein significantly correlated with contractile potency for these agonists in human cerebral arteries. Thus, to the extent that contractile responsiveness of human cerebral arteries may predict antimigraine agents, contractile responses of the rabbit and/or canine saphenous vein may be useful surrogates for antimigraine efficacy. In addition, the contractile potency for this series of serotonergic agonists in the rabbit or canine saphenous vein significantly correlated with contractile potency of these agonists in human coronary arteries. These data suggest that the use of the saphenous vein to identify potent vasoconstrictors will also reveal agents capable of contracting human coronary arteries, a liability for using this approach to evaluate promising antimigraine therapies. PMID- 9350383 TI - Peripheral vascular effects and pharmacokinetics of the antimigraine compound, zolmitriptan, in combination with oral ergotamine in healthy volunteers. AB - Members of the new class of antimigraine compounds, 5HT1B/1D agonists, as well as ergotamine, may cause vasoconstriction through stimulation of 5HT receptors on peripheral vessels. The cardiovascular effects of 20 mg oral zolmitriptan (Zomig, formerly 311C90), 2 mg oral ergotamine and the combination were assessed in a randomized double-blind, placebo-controlled crossover study in 12 healthy subjects. Pharmacodynamic measures included oscillometric blood pressure, systolic blood pressure at the toe and arm using a strain gauge technique, stroke volume and cardiac output using bioimpedance cardiography, high-resolution ultrasound to measure brachial arterial diameter and a novel Doppler method to measure blood flow velocity. Both drugs produced small degrees of peripheral vasoconstriction, including increases in diastolic blood pressure and blood flow velocity and decreases in arterial diameter and toe-arm systolic pressure gradient. These effects were generally additive with the combination but of no clinical importance. There were no significant changes in cardiac output, stroke volume heart rate or ECG. Zolmitriptan, at eight times the likely therapeutic dose, was generally well tolerated both alone and in combination with ergotamine. Ergotamine had no clinically important effects on zolmitriptan pharmacokinetics. PMID- 9350384 TI - Double-blind, placebo-controlled, dose-finding study of rizatriptan (MK-462) in the acute treatment of migraine. AB - Rizatriptan (MK-462) is a potent 5HTID receptor agonist. This multicenter, double blind, placebo-controlled, outpatient study investigated the clinical efficacy, safety, and tolerability of rizatriptan (2.5, 5, and 10 mg) as a function of dose for acute migraine. Patients with moderate or severe migraine (n = 417) were treated with placebo (n = 67), rizatriptan 2.5 mg (n = 75), 5 mg (n = 130), or rizatriptan 10 mg (n = 145). Headache severity, functional disability, and migraine symptoms were measured immediately before dosing (0) and at 0.5, 1, 1.5, 2, 3, and 4 h post-dose. Patients were permitted to take a second dose of test drug at 2 h if their headache pain was moderate or severe (i.e., placebo initially-->rizatriptan 10 mg as optional second dose; rizatriptan 2.5 mg, 5 mg, or 10 mg initially-->placebo as optional second dose). An upward dose-response relationship was observed among placebo, rizatriptan 2.5 mg, 5 mg, and 10 mg in the primary efficacy measure of proportion of patients reporting pain relief, i.e., a change in headache severity to "no pain or mild pain" at 2 h post-dose. The relationship was evident even at the first recorded timepoint, 30 min, and was statistically significant at 1.5 h and beyond. At the primary timepoint of 2 h after the initial dose, the proportion of patients reporting pain relief was 47.6% for rizatriptan 10 mg; 45.4% for rizatriptan 5 mg; 21.3% for rizatriptan 2.5 mg; and 17.9% for placebo. Seventy percent of patients on rizatriptan 10 mg reported pain relief at 4 h. Patients who took rizatriptan 5 mg and 10 mg were significantly less functionally disabled than those who took placebo at 1.5 and 2 h post-dose. Rizatriptan 10 mg was consistently more effective than 5 mg, although the differences were not statistically significant. The most frequent clinical adverse events were dizziness, somnolence, and asthenia/fatigue. No patients were discontinued for any adverse experiences and there were no serious adverse experiences. PMID- 9350385 TI - Erythrocyte and plasma levels of glutamate and aspartate in children affected by migraine. AB - In this study we determined plasma and erythrocyte amino acids in children affected by migraine, in order to evaluate glutamate and aspartate metabolism in the pathogenesis of this disorder. Fifteen children with migraine with aura (mean age +/- SD = 10.3 +/- 1.56), 19 children with migraine without aura (mean age +/- SD = 10.4 +/- 1.48) and 16 healthy normal controls (mean age +/- SD 10.6 +/- 1.53) were investigated. In both migraine groups there were significantly lower plasma glutamate and aspartate levels and significantly higher erythrocyte/plasma concentration (E/P) ratios of these amino acids with respect to the controls. Erythrocyte aspartate concentrations were significantly elevated in migraine children compared to the controls, while erythrocyte glutamate concentrations showed no significant differences between groups. Similar results were observed in both migraine groups. These results seem to suggest the presence of a higher activity of the erythrocytes' glutamate/aspartate transport system that could reflect a similar alteration at the neuronal/glial cell level in the CNS. Our study suggests an imbalance of the excitatory amino acid turnover in the pathogenesis of migraine in children. PMID- 9350386 TI - Endozepine stupor in children. AB - Recurring episodes of stupor in adults have been shown to be related to increased levels of endozepines, which are endogenous ligands for the GABAA receptors. We report here two children presenting with recurrent episodes of stupor associated with fast EEG activity who had increased levels of endozepine-4 in plasma. Mass spectroscopy did not reveal commercially available benzodiazepines. Interictal endozepine-4 levels were normal. In one of the patients, administration of flumazenil (0.25 mg i.v.), a benzodiazepine inverse agonist, induced improvement of consciousness and attenuation of EEG fast activity. In conclusion, children presenting with recurrent episodes of stupor and EEG fast activity should be evaluated for endozepine levels and can be effectively treated with i.v. flumazenil. PMID- 9350387 TI - Outcome of early school-age migraine. AB - We studied the outcome of migraine from early school-age to prepuberty in a group of 84 children. The children belonged to a population-based, unselected follow-up sample of a 1-year age cohort. At the age of 8 to 9 years, 95 (2.7%) children of this age cohort had migraine according to a postal questionnaire. At age 11 to 12 years, 84 of them were traced and interviewed face-to-face. Only four (4.8%) of these children no longer had headache. Fifty-three (63.1%) children had migraine. Seventeen (20.2%) had migraine-type headache which did not completely fulfil the International Headache Society criteria for migraine, seven (8.3%) children had episodic tension-type headache and three (3.6%) had other headache. Among the children who had migraine at age 11 to 12, boys had significantly more frequent migraine attacks than girls (mean 2.7/month versus 1.8/month; p = 0.016). They also used more drugs and were more frequently absent from school because of headache than girls, but these differences were not significant. Problems in the relationships between parents seemed to be another factor associated with frequent migraine. PMID- 9350388 TI - The m-chlorophenylpiperazine test in cluster headache: a study on central serotoninergic activity. AB - The central serotoninergic agonist m-chlorophenylpiperazine (m-CPP) stimulates several 5HT receptor subtypes. It induces the release of both cortisol and prolactin (PRL). In this study we investigated central serotoninergic responsiveness in cluster headache by monitoring cortisol and PRL responses to m CPP administration. Twenty-three patients with episodic cluster headache and 17 sex-matched and age-matched healthy subjects were studied. The cluster headache patients were tested during a cluster period, and none were receiving prophylaxis. A single oral dose of m-CPP, 0.5 mg/kg, was given at time 0. Blood samples were drawn at -30, 0, 30, 60, 90, 120, 150 and 180 min. PRL and cortisol levels were assayed in the samples. PRL and cortisol delta maxima (delta maximum = maximum response - baseline level at time 0/baseline level at time 0) were evaluated in each patient and mean values compared. Serum levels of m-CPP were detected by HPLC and correlated to hormonal responses. Reduced cortisol (p < 0.02) and increased PRL (p < 0.05) delta maxima were observed in cluster headache patients. Increased basal cortisol plasma levels (p < 0.05) and reduced basal PRL plasma levels (p = 0.06) also characterized cluster headache patients. This is the first study evaluating central serotoninergic responsiveness to m-CPP in cluster headache and these data suggest impaired central serotoninergic function in this pathology. PMID- 9350389 TI - Double-blind placebo-controlled trial of lithium in episodic cluster headache. AB - Lithium is widely used in the prophylaxis of episodic cluster headache without formal evidence of efficacy. Placebo-controlled clinical trials are not easy in conditions characterized by frequent severe pain. In this study, it was assumed that lithium would work quickly if at all, and placebo response would be zero. Strict diagnostic criteria excluded uncertain or atypical cases. Patients were male in so-far untreated episodes expected to last for at least 3 weeks more. In a double-blind, placebo-controlled comparison of matched parallel groups, treatment was either slow-release lithium carbonate, 800 mg/day, or placebo. After 7 days, compliance was estimated by tablet count, blood was taken for lithium assay, efficacy was assessed (attacks stopped or substantially improved) and adverse reactions were recorded. The study was stopped after planned sequential analysis of the 27th patient (13 on lithium, 14 on placebo). Estimated compliance was usually but not always good. Plasma lithium levels were mostly in the range 0.5-0.6 mmol/l on lithium, zero on placebo. Cessation of attacks within 1 week occurred in two patients in each group, substantial improvement in 6/14 (43%) on placebo, 8/13 (62%; NS) on lithium. Only minor adverse events were reported. Lithium treatment was therefore associated with a useful subjective improvement rate but the assumptions made at outset had proved wrong. The trial was stopped because superiority over placebo could not be demonstrated. There were lessons for future trials. PMID- 9350390 TI - Orbito-sphenoidal Aspergillus infection mimicking cluster headache: a case report. PMID- 9350391 TI - Cluster headache after orbital exenteration. AB - A 37-year-old man developed an ipsilateral headache which fulfilled the criteria for cluster headache after orbital extenteration because of a traumatic lesion of the bulb. The headache could be treated successfully by drugs usually applied in the therapy of cluster headache. Six similar cases of cluster headache after orbital exenteration could be identified in the literature suggesting that the eye itself is not necessarily part of the pathogenesis of cluster headache. We hypothesize that orbital exenteration can cause cluster headache by lesions of sympathetic structures. Possibly, these mechanisms are similar to those of sympathetic reflex dystrophy (Sudeck-Leriche syndrome) causing pain of the limbs. PMID- 9350392 TI - Genetic epidemiology of migraine and cluster headache. PMID- 9350393 TI - Organization and delivery of services to headache patients. Task Force of The International Headache Society. PMID- 9350394 TI - Central sites of actions of zolmitriptan and sumatriptan: 5HT1B, 5HT1D and 5HT1F receptor distribution. PMID- 9350395 TI - Antigliadin antibodies in migraine patients. PMID- 9350396 TI - Hereditary epilepsy syndromes. AB - This paper reviews the present knowledge on the genetics of the epilepsies. Main clinical features, gene localization and pattern of inheritance of the idiopathic epilepsies, the progressive myoclonus epilepsies, and some other genetic disorders often associated with epilepsy, are described. PMID- 9350397 TI - Hyperekplexia-like syndromes without mutations in the GLRA1 gene. AB - Hyperekplexia (MIM: 149400), or startle disease, is an autosomal dominant neurological disorder characterized by an extreme generalized stiffness immediately after birth, normalizing during the first years of life. Other features of this disorder are excessive startle reactions to unexpected, particularly auditory, stimuli together with a short period of generalized stiffness during which voluntary movements are impossible. Linkage analysis mapped a gene for this disorder to chromosome 5q33-q35. Subsequently, mutations in the GLRA1 gene encoding the alpha 1-subunit of the glycine receptor proved to be causally related to the disease. In the present study, mutation analysis of all exon and flanking intron sequences of this gene was performed in sporadic patients and their parents. Moreover, a branch of the original Dutch hyperekplexia family with a very severely affected individual was screened for an additional mutation in the GLRA1 gene. Except for two polymorphisms, of which one results in an amino acid change, no potentially disease causing mutations were found in the alpha 1-subunit of the glycine receptor. Together with haplotype analysis these results exclude a recessive inheritance or new mutation etiology in these hyperekplexia-like syndrome and emphasize that hyperekplexia-like syndromes can be caused by other genetic factors. The involvement of other genes encoding subunits of the functional glycine receptor complex has not been excluded. PMID- 9350398 TI - Supratentorial cavernous malformations and epilepsy: seizure outcome after lesionectomy on a series of 35 patients. AB - Epilepsy is the most frequent presenting sign in patients with cavernous angiomas and is the major cause of morbility. Persistence of seizures after surgical treatment prompted many authors to examine the possibility of removing the cavernoma and the surrounding tissue. In our series of 53 cavernous angiomas, all the 35 patients with preoperative seizures underwent surgery by means of lesionectomy alone. One hundred percent of patients with less than five preoperative seizures and/or an history under 12 months was seizure free, while only 62.5% of patients with more than five seizures and/or an history longer than 12 months was seizure free. Number and duration of seizures before surgery seems to be the most important factor in the seizure outcome after surgical treatment. PMID- 9350399 TI - Molecular aspects of neuro-oncology. AB - Detailed understanding of molecular events responsible for brain tumor growth is a prerequisite for the development of effective therapeutic modalities leading to improved prognosis and cure. Advances in molecular biology in the past decades have revolutionized our understanding of cancer, including brain tumors. We have learned that abnormal proliferation, inability of the cells to die and their potential to modify their tissue environment result from accumulation of genetic aberrations. This article reviews genetic mechanisms implicated in the pathogenesis of nervous system tumors, such as unactivation of tumor suppressor and replication error genes, generation of abnormal growth factor loops, alterations of apoptotic pathways and angiogenesis. PMID- 9350400 TI - Glossopharyngeal schwannoma, an uncommon posterior fossa tumor: diagnostical and therapeutical aspects: a case report. AB - Among posterior fossa tumors, schwannomas arising from glossopharyngeal nerve are rare; only about 30 cases of glossopharyngeal neuroma have been described. Though a typical 'jugular foramen syndrome' has been described for tumors of this region, the clinical onset may often closely resemble that of acoustic neuromas thus misleading the diagnosis. Because of its different surgical implications, such a rare condition must be clearly recognised preoperatively. A new case is hereby reported with particular focus on clinical and surgical management. PMID- 9350401 TI - Spontaneous intracranial hypotension: clinical and magnetic resonance imaging characteristics. AB - The syndrome of spontaneous intracranial hypotension (SIH) is an uncommon cause of postural headache. We describe three patients with classical low pressure headache associated with low CSF pressure, one of whom presented with sudden deafness and another with a unilateral VIth nerve palsy. Initial magnetic resonance imaging (MRI) scans revealed bilateral diffuse subdural fluid collections in all three cases. Follow up MRI scans performed on two patients at 6 months demonstrated partial resolution of the subdural collections but persistent striking meningeal enhancement despite clinical recovery. These findings differ from previous reported cases wherein clinical resolution of postural symptoms was preceded or closely followed by resolution of the MRI changes. PMID- 9350402 TI - Epidural electrical stimulation of the motor cortex in patients with facial neuralgia. AB - Chronic facial neuralgias often do not respond sufficiently to standard treatment methods. Alternative modalities are needed for long-term reduction of pain in such cases. The present preliminary report describes two patients with trigeminal and glossopharyngeal neuralgia, respectively, treated with standard methods without obtaining satisfactory pain relief. Electrical stimulation of the motor cortex contralateral to the pain area was employed in both cases and proved able to produce a long-term facial pain reduction. Alleviation of pain occurred after activation of the flat quadripolar electrode placed epidurally on the precentral cortical area and lasted as long as the stimulator was working. By changing the polarity of the electrodes, it was possible to induce tingling sensations and muscle activation not only contralaterally to the stimulated motor cortex, but also in the ipsilateral part of the face. No stimulator-independent pain reduction resulted from long-term use of the stimulation device. During a follow up period of 18 months, a sufficient and relatively stable analgesic effect of electrostimulation was observed. One major complication of motor cortex stimulation during the follow-up period was a single generalized epileptic seizure in one of the patients. PMID- 9350403 TI - Primary cerebral leiomyosarcoma. AB - A case of a rare primary cerebral leiomyosarcoma in an 8-year-old male is described. The patient presented with a new-onset seizure disorder and was found to have a rapidly expanding left parietal extra-axial lesion, documented by radiological imagings. The patient underwent surgical resection of the leiomyosarcoma, as well as adjuvant radiotherapy and chemotherapy. He is still surviving to date in stable neurological condition. The clinical presentation, surgical procedure, pathological findings and post-operative clinical course will be reported. The possible etiology of this rare extra-axial neoplasm is discussed. PMID- 9350404 TI - Regional cerebral blood flow changes related to affective speech presentation in persistent vegetative state. AB - A story told by his mother was presented on tape to a trauma patient in persistent vegetative state (PVS). During auditory presentation, measurements of regional cerebral blood flow (rCBF) were performed by means of positron emission tomography (PET). Changes in rCBF related to this stimulus condition, as compared to presenting non-word sound, were evaluated by means of statistical parametric mapping (SPM). This analysis indicated activation of rostral anterior cingulate, right middle temporal and right premotor cortices, which may reflect appropriate cortical involvement in processing emotional attributes of sound or speech. PMID- 9350405 TI - Hemiplegia caused by inadvertent intra-carotid infusion of total parenteral nutrition. AB - A 24 year-old woman developed acute hemiplegia and a seizure following accidental catheterization of the right common carotid artery and total parenteral nutrition infusion. Magnetic resonance imaging of the brain showed lesions in the frontal lobe and putamen consistent with an ischemic stroke. Angiography through the central venous catheter confirmed its intra-arterial location. The patient's weakness improved after hyperbaric oxygen treatment. We concluded that stroke or seizures during total parenteral nutrition administration through a central venous catheter should alert one to the possibility of inadvertent intra-arterial infusion, especially in patients who have had central lines inserted several times previously. PMID- 9350406 TI - A rare type of false negative three-dimensional CT angiography of a cerebral aneurysm. PMID- 9350407 TI - Reversible posterior leukoencephalopathy following organ transplantation. Description of two cases. AB - Although neurologic changes after organ transplantation are often secondary to opportunistic infections or vascular insults, new pathological entities are emerging. We have recently encountered two patients who, a few days after liver and heart transplant, respectively, developed neurological signs and symptoms. Head computerized tomography (CT) scan showed nonenhancing areas of low attenuation, and magnetic resonance imaging (MRI) demonstrated multiple areas of increased signal intensity in the subcortical white matter on T2-weighted images. Stereotactic biopsy of the intracranial lesions was performed in one case. Light microscopic examination demonstrated only mildly edematous white matter. No infectious organisms were observed on light or electron microscopy. In one patient, follow-up MRI 3 months later showed almost complete resolution of the signal abnormalities. Both patients' clinical condition progressively improved. The neuroradiological abnormalities described are consistent with the 'reversible posterior leukoencephalopathy' syndrome associated with cyclosporine toxicity. The pathophysiology of these lesions is unclear; however, it has been suggested that cyclosporine causes an acute ischemic insult secondary to vascular spasm with resultant axonal swelling. This hypothesis is supported by the hypoattenuation seen on CT, the prolonged T2 relaxation seen on MRI, and the absence of contrast enhancement. Concomitant factors (such as hypocholesterolemia or associated therapy with high dose steroids) are important in the development of these lesions as in both of our patients cyclosporine levels were in the normal range. Fortunately, these lesions and the associated manifestations are most often reversible and regress with adjustments of cyclosporine dosage and/or correction of concomitant facilitating factors. PMID- 9350408 TI - Comparison of cycloserine-cefoxitin-fructose agar (CCFA) and taurocholate-CCFA for recovery of Clostridium difficile during surveillance of hospitalized patients. AB - The effectiveness of cycloserine-cefoxitin-fructose agar (CCFA) and taurocholate CCFA (TCCFA) in isolating Clostridium difficile from swabs of the rectum or stools from 184 hospitalized patients who were monitored weekly and when they had diarrhea was compared. The number of surveillance time points ranged from two to eight per patient over a period of 4 to 34 days per patient, totalling 621 comparisons of the media. C. difficile was isolated more frequently by TCCFA than CCFA at seven of eight surveillance points, a significant trend (O'Brien test, p = 0.002). This difference reached statistical significance at the second surveillance time point when the prevalence of C. difficile was sufficiently high. At the second surveillance point, C. difficle was isolated only by TCCFA in 7 of 184 comparisons of the media, only by CCFA in none of the comparisons, and by both media in 19 comparisons (p = 0.016). C. difficle was first isolated at an earlier surveillance time point on TCCFA in 11 of 36 patients and on CCFA first only once (p = 0.005). Use of TCCFA media increased the rapidity and sensitivity of culture for C. difficle when doing patient surveillance but did not increase sensitivity when diagnosing patients with diarrhea. PMID- 9350409 TI - Candidemia in a Canadian tertiary care hospital from 1976 to 1996. AB - The incidence of candidemia was reviewed at the Health Sciences Centre in Winnipeg, Canada, over a 21-year period (1976 to 1996). Candida species were identified as blood-stream isolates in significantly (p < 0.05) higher numbers from 1991 to 1996 than in the previous 15 years. Antifungal susceptibilities remained unchanged with Candida albicans isolates tested from 1985 to 1996. Retrospective chart reviews revealed that all patients with candidemia possessed at least two risk factors. The main risk factors identified were recent or concurrent antibiotic therapy (95% of patients), presence of a central line (93% of patients), and immunosuppression (88% of patients). Treatment generally involved amphotericin B therapy (81% of patients), and death occurred in 52% of the patients. Mortality directly attributable to Candida species could be established in 23% of patients. PMID- 9350410 TI - Molecular typing and fluconazole susceptibility of urinary Candida glabrata isolates from hospitalized patients. AB - At our community teaching hospital between August 1994 and August 1995, Candida glabrata accounted for 14% of all Candida isolates and for 31% of urinary Candida isolates. The culture site was urine for 68% of C. glabrata isolates compared to 30% of all Candida isolates (p < 0.001, chi 2). To study the association between C. glabrata and isolation from the urine, we analyzed all available C. glabrata urinary isolates over a 3-month period (23 isolates from 20 patients) using electrophoretic karyotyping, random amplified polymorphic DNA analysis, and fluconazole susceptibility testing. Random amplified polymorphic DNA generated eight types, although electrophoretic karyotyping generated 17 types. Combining the two methods resulted in 19 types indicating that urinary C. glabrata strains at our hospital are genetically diverse and the association between C. glabrata and urinary tract isolation does not appear to be due to horizontal transmission of a single or small number of strains. In vitro susceptibility tests showed that C. glabrata isolates from patients receiving fluconazole had significantly higher minimum inhibitory concentrations to fluconazole than those not receiving fluconazole (p < 0.05). Despite a limited number of patients and isolates, our data suggest that selection of less susceptible organisms by the presence of antifungal agents may be an important contributor to increased urinary isolation of C. glabrata from patients in our hospital. PMID- 9350411 TI - Use of different PCR-based DNA fingerprinting techniques and pulsed-field gel electrophoresis to investigate the epidemiology of Acinetobacter calcoaceticus Acinetobacter baumannii complex. AB - Acinetobacter calcoaceticus-Acinetobacter baumannii complex is an important nosocomial pathogen for which optimal typing methods in epidemiologic investigations have not been defined. We compared DNA macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) with different PCR-based DNA fingerprinting techniques, including enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR), repetitive extragenic palindromic (REP) PCR, arbitrary-primed PCR with primer M13, and multiplex PCR with primers REP-1, REP-2 and M13, for characterization of 98 clinical isolates (including 10 apparent outbreak-related isolates and 68 presumed epidemiologically unrelated isolates) in a tertiary-care hospital over a 4-year period. The PFGE patterns after Smal restriction of the bacterial DNA were analyzed by computer software (Gelcompar) using the unweighted pair group method with arithmetic averages clustering and the Dice coefficient. A cluster of 48 isolates (cluster A), including 9 outbreak isolates, linked at a level of 83.4% similarity was observed. This epidemic strain and its variants were also found among the 68 presumed epidemiologically unrelated isolates, and this may represent ongoing endemic infection in this institution. The discrimination index for the PCR-based DNA fingerprinting techniques was 0.75 for enterobacterial repetitive intergenic consensus 1, 0.71 for M13, 0.77 for REP-1, 0.77 for REP-2, and 0.87 for multiplex PCR. The discriminatory power of PFGE was found to be higher than those of PCR-based techniques. It was concluded that both PFGE and PCR-based fingerprinting are useful for typing of A. calcoaceticus-A. baumannii complex. However, PFGE can detect minor mutations among outbreak strains, and this is important for epidemiological study of this species in a complex endemic setting. PMID- 9350412 TI - Argentinian collaborative study on prevalence of erythromycin and penicillin susceptibility in Streptococcus pyogenes. The Argentinian Streptococcus Study Group. AB - Two monthly studies on the prevalence of penicillin and erythromycin susceptibility of Streptococcus pyogenes were performed in May and October of 1994 in Argentina. A total of 58 centers from 27 cities participated in these studies. A total of 1072 isolates were tested by a diffusion method, although 595 isolates were tested both by the diffusion and an agar dilution method (n = 1767 isolates). No penicillin-resistant streptococci were found in our study (MIC100 = 0.03 microgram/ml). Only four isolates were confirmed as erythromycin resistant S. pyogenes (prevalence 0.14 and 0.28% in May and October 1994, respectively). Resistance in three isolates was due to an inducible mechanism, although in one strain a different phenotype was observed. PMID- 9350413 TI - In vitro activity of trimethoprim alone compared with trimethoprim sulfamethoxazole and other antimicrobials against bacterial species associated with upper respiratory tract infections. AB - Trimethoprim-sulfamethoxazole has been used to treat various respiratory tract infections. Nevertheless, for many patients, intolerance of the sulfonamide component precludes use of this combination. This study examined the activity of trimethoprim alone in comparison to that of trimethoprim-sulfamethoxazole and other antimicrobials against bacterial species implicated in respiratory tract infections. For Haemophilus influenzae, minimal inhibitory concentrations of trimethoprim were equal to or one dilution greater than those of trimethoprim sulfamethoxazole, with 56 of 58 strains inhibited by the former at < or = 0.25 microgram/ml. All oxacillin-susceptible Staphylococcus aureus and 96.7% of Streptococcus pyogenes were inhibited by trimethoprim < or = 2 micrograms/ml. In contrast, only 50% of Streptococcus pneumoniae were inhibited by this concentration of trimethoprim, whereas 93.3% were susceptible to the combination at < or = 2/38 micrograms/ml. All oxacillin-resistant S. aureus and all Moraxella catarrhalis were resistant to trimethoprim, although many of the former and all of the latter were susceptible to trimethoprim-sulfamethoxazole. PMID- 9350414 TI - High rates of resistance to piperacillin/tazobactam among Escherichia coli and Klebsiella pneumoniae strains isolated in a Greek hospital. AB - The activities of piperacillin/tazobactam (PTZ) and 10 other beta-lactams were evaluated by a broth microdilution method using the PASCO system in 1078 Escherichia coli and 447 Klebsiella pneumoniae strains isolated from hospitalized patients in a Greek hospital. Overall, 10.5% of the former and 62.6% of the latter strains displayed resistance to PTZ. Most of the PTZ-resistant strains (71.2%) expressed extended spectrum beta-lactamases and were also resistant to broad spectrum cephalosporins and aztreonam. The high rates of PTZ resistance in E. coli and K. pneumoniae and the extensive cross-resistance to other beta lactams suggest that PTZ should be used with caution in our clinical setting. PMID- 9350415 TI - Vancomycin-resistant Enterococcus raffinosus: molecular epidemiology, species identification error, and frequency of occurrence in a national resistance surveillance program. AB - Enterococcal blood stream infections are the third most common among all nosocomial blood stream infections in the United States and the occurrence of glycopeptide (vancomycin, teicoplanin) resistance in these isolates has markedly increased. Control of hospital-acquired infections with vancomycin-resistant enterococci requires high quality antimicrobial susceptibility test methods and species identification procedures as a supplement to epidemiologic investigation and appropriate infection control procedures. In this report, bacteremias caused by Enterococcus avium (BioMerieux Vitek, Hazelwood, MO, USA) were observed to be Enterococcus raffinosus infections (six of eight cases; 1.1% of all cases) when reference biochemical identification methods were applied. The vancomycin susceptible E. raffinosis (two strains) and E. avium (two strains) had unique phenotypic and genotypic molecular profiles. In contrast, four vancomycin resistant E. raffinosus strains (van A by polymerase chain reaction) from a single institution had the same phenotypic and molecular (PCR, PFGE, ribotyping) pattern, indicating clonal dissemination among four patients over a 66-day period. Clinical laboratories should be aware of the high probability that van A genes may be transferred from Enterococcus faecium or Enterococcus faecalis to other more rarely encountered Enterococcus species. Also contemporary, widely used commercial identification systems may fail to accurately identify those rare species. Errors appear to be most prevalent for E. avium, Enterococcus durans, and E. raffinosus based on the experience of the SCOPE Program. PMID- 9350416 TI - Fusospirochetal superinfection of pre-existing oral lesion in patients with acquired immunodeficiency syndrome. AB - Three patients with AIDS presented with nonbleeding, painful, fetid, oral ulcers overlaid with a grayish-black semiadherent membrane at the sites of a pre existing lesion. These lesions persisted despite treatment directed toward the primary etiology (cytomegalovirus, Kaposi's sarcoma). Gram- and Giemsa-stained smears of teased membrane fragments revealed an impressive bacterial flora with fusiforms and Borrelia-type spirochetes. Prompt treatment with penicillin brought amelioration of symptoms and sloughing of the overlaying membrane. PMID- 9350417 TI - Anti-Legionella activity of trovafloxacin compared with seven other antimicrobial agents including an intermethod evaluation. AB - The activity of trovafloxacin, a new fluorinated naphtheridone, was tested against 61 Legionella spp. isolates and compared with that of 4 fluoroquinolones, 2 macrolides, and rifampin. Trovafloxacin MICs were determined by a reference agar dilution method and E-test (Solna, Sweden) strips on buffered charcoal yeast extract agar. Among the fluoroquinolone compounds, the rank order of activity (on the basis of MIC90 results) determined with E-test strips was as follows: levofloxacin (MIC90, 0.094 microgram/ml) > trovafloxacin = sparfloxacin = ofloxacin (MIC90, 0.19 microgram/ml). Rifampin (MIC90, 0.008 microgram/ ml) and clarithromycin (MIC90, 0.032 microgram/ml) were the most potent of all drugs tested, and erythromycin and ciprofloxacin were the least active. In this study, the E-test strips with trovafloxacin were validated (100% of results +/- one log2 dilution compared to the reference value) for susceptibility testing with Legionella isolates. PMID- 9350418 TI - Ionizing radiation induces, via generation of reactive oxygen intermediates, intercellular adhesion molecule-1 (ICAM-1) gene transcription and NF kappa B-like binding activity in the ICAM-1 transcriptional regulatory region. AB - Ionizing radiation produces reactive oxygen intermediates in mammalian tissues and may serve as a model system for the investigation of the biologic effects of free radicals. We have previously shown that the adhesion molecule ICAM-1 is induced by ionizing radiation, and here we have investigated the molecular mechanisms responsible. ICAM-1 mRNA and cell surface expression was induced in HeLa and HaCaT cells after exposure to ionizing radiation. This induction was blocked by preincubation with the antioxidants PDTC and N-acetyl cysteine. ICAM-1 promoter activity was assessed by transiently transfecting HeLa cells with CAT reporter gene constructs containing sequential ICAM-1 5' deletions. ICAM-1 5' fragments -1162/+1 (relative to the transcription start site) and -277/+1 displayed increased promoter activity when cells were exposed to ionizing radiation, but no induction was seen in a -182/+1 construct associating positions -277 to around -182 with inducibility by ionizing radiation. Nuclear extracts from HaCaT cells were tested in mobility shift assays using an NF kappa B-like binding site of the ICAM-1 5' region (positions -186/-177). There was marked enhancement of DNA-protein complex forming in extracts from irradiated versus untreated cells. Incubation of cells with antioxidants prior to irradiation prevented the radiation-dependent increase in complex formation. We conclude that reactive oxygen intermediates are involved in ICAM-1 induction by ionizing radiation. The ionizing radiation-induced, antioxidant-inhibitable binding at the ICAM-1 NF kappa B-like binding site is consistent with the view that NF kappa B is a pro-oxidant transcription factor. PMID- 9350419 TI - Diethyldithiocarbamate and nitric oxide synergize with oxidants and with membrane damaging agents to injure mammalian cells. AB - The effect of diethyldithiocarbamate (DDC) and sodium nitroprusside (SNP) on the killing of endothelial cells and on the release of arachidonate by mixtures of oxidants and membrane-damaging agents was studied in a tissue culture model employing bovine aortic endothelial cells labeled either with 51Chromium or 3arachidonic acid. While exposure to low, subtoxic concentrations of oxidants (reagent H2O2, glucose-oxidase generated peroxide, xanthine xanthine oxidase, AAPH-generated peroxyl radical, menadione-generated oxidants) did not result either in cell death or in the loss of membrane-associated arachidonic acid, the addition of subtoxic amounts of a variety of membrane-damaging agents (streptolysin S, PLA2, histone, taurocholate, wheatgerm agglutinin) resulted in a synergistic cell death. However, no significant amounts of arachidonate were released unless proteinases were also present. The addition to these reaction mixtures of subtoxic amounts of DDC (an SOD inhibitor and a copper chelator) not only very markedly enhanced cell death but also resulted in the release of large amounts of arachidonate (in the complete absence of added proteinases). Furthermore, the inclusion in DDC-containing reaction mixtures of subtoxic amounts of SNP, a generator of NO, further enhanced, in a synergistic manner, both cell killing and the release of arachidonate. Cell killing and the release of arachidonate induced by the DDC and SNP-containing mixtures of agonists were strongly inhibited by catalase, glutathione, N-acetyl cysteine, vitamin A, and by a nonpenetrating PLA2 inhibitor as well as by tetracyclines. A partial inhibition of cell killing was also obtained by 1,10-phenanthroline and by antimycin. It is suggested that DDC might amplify cell damage by forming intracellular, loosely bound complexes with copper and probably also by depleting antioxidant thiols. It is also suggested that "cocktails" containing oxidants, membrane-damaging agents, DDC, and SNP might be beneficial for killing of tumor cells in vivo and for the assessment of the toxicity of xenobiotics in vitro. PMID- 9350420 TI - Absence of synproportionation between oxy and ferryl leghemoglobin. off. AB - The synproportionation reaction between ferryl leghemoglobin and oxyleghemoglobin does not occur, at least under conditions where this process could be clearly demonstrated with myoglobin and hemoglobin. In contrast, a cross synproportionation can occur between oxyleghemoglobin and ferryl myoglobin or between ferryl leghemoglobin and oxymyoglobin. The non-exposure, at the surface of the leghemoglobin molecule, of the nearest tyrosine residue to the heme group could explain this behaviour. Thus leghemoglobin per se does not appear to be able to act as an antioxidant in removing H2O2 by synproportionation. However, in the presence of ascorbate and/or glutathione which can reduce ferryl leghemoglobin, this hemoprotein could act as an H2O2-removing antioxidant, in a process similar to that described for myoglobin. This could also explain why, despite the absence of synproportionation, ferryl leghemoglobin is not detected in nodule extracts. PMID- 9350421 TI - Primaquine alters antioxidant enzyme profiles in rat liver and kidney. AB - The effects of primaquine treatment on antioxidant enzyme activities were investigated in rat liver and kidney. Male Sprague-Dawley rats were treated with 0.21 mg/kg daily for two weeks (chronic treatment) or a single dose at 0.21 or 0.63 mg/kg. Antioxidant enzyme activities were determined in liver and kidney cytosolic fractions whereas glutathione (GSH) and malondialdehyde (MDA) levels were determined in tissue samples. Results for the liver showed increases in cytosolic superoxide dismutase (SOD) and glutathione peroxidase (GPX) enzymatic activities after chronic primaquine treatment. Levels of MDA, a marker for lipid peroxidation, were also increased by more than 50% indicating enhanced oxidative damage in the liver. In the single dose study, 0.63 mg/kg primaquine caused a more than 100% increase in liver SOD and a 36% increase in NAD (P) H: quinone oxidoreductase (NQOR) activities. Results for the kidney, however, showed fewer primaquine-induced changes in antioxidant enzyme activities when compared to the liver in both the chronic and single dose studies. Overall, our results indicate that primaquine treatment causes an oxidative stress in the two rat organs. These results are consistent with the known pro-oxidant effects of primaquine in vivo, and supplement current knowledge on the effects of antimalarial drugs on various enzyme systems. PMID- 9350422 TI - Radical formation from the reaction of combustion smoke with diphenylamine. AB - We have attempted to examine the effects of radical scavengers, such as amines and phenols, to trap gas-phase radicals produced from the combustion of Poly (methyl methacrylate)(PMMA), which might cause damage to a living body, using an electron spin resonance (ESR) spin-trapping technique. As a result, diphenylamine did not decrease the amount of radicals but rather increased it. It indicates that under the conditions of this study, gas-phase radicals were hardly trapped by radical scavengers and that the precursors to produce other kinds of radicals can exist. It was suggested that from the experiments using several peroxides, the precursors should be diacylperoxides produced from the combustion of PMMA. PMID- 9350423 TI - Effects of alpha-phenyl-N-tert-butyl nitrone (PBN) on compression injury of rat spinal cord. AB - alpha-Phenyl-N-tert-butyl Nitrone (PBN) is a free radical scavenger which recently has proved to be neuroprotective in experimental studies on focal cerebral ischemia and infarction. We therefore studied the effect of this drug in a model of moderate compression injury to rat spinal cord at the midthoracic level. The compound was given intraperitoneally 0.5 h before (100 mg/kg b.w) and at 1.5 h (50 mg/kg b.w) and 3.5 h (50 mg/kg b.w) after compression. Treated animals and controls (vehicle alone) were allowed to survive for 1 or 9 days following trauma. The functional outcome was tested by the inclined plane method and the motor performance score. By using MAP2 immunostaining the number of nerve cell bodies in the ventral horn and the ratio of MAP2 immunostained area to area of whole section of the cord were assessed to detect loss of neurons and loss of dendrites in the compressed segment. beta APP and PGP9.5 immunostaining was used to demonstrate axonal lesions. Treated and control rats showed at day 1 when tested with the inclined plane method a marked reduction of the capacity angle. This abnormality recovered gradually over the following days and was normalized at day 9. The motor performance score showed a marked reduction at day 1 which almost normalized at day 9. There was no difference regarding the functional outcome between rats given PBN and controls in none one of these functional tests. The spinal cord of normal rats presented immunoreactivity to MAP2 in nerve cell bodies and dendrites but not in axons and other structures. Following compression there was at day 1 and 9 a marked loss of MAP2 immunoreactivity in dendrites and nerve cell bodies. We could not detect any difference between the PBN and the control rats regarding the degree of cell loss or degree of reduction of dendrite staining. No difference between the two groups was seen with the axonal immunostainings (beta APP and PGP9.5). In conclusion, our study did not reveal any neuroprotective effect of PBN on the functional outcome and morphology (immunostaining to MAP2, beta APP and PGP9.5) in this model of moderate compression trauma to rat spinal cord. PMID- 9350424 TI - New method to measure the carbamoylating activity of nitrosoureas by electron paramagnetic resonance spectroscopy. AB - A new method for measuring the carbamoylating activity of nitrosoureas and isocyanates using electron paramagnetic resonance (EPR) spectroscopy is described. The extent and time course of carbamoylation reaction of chloroethyl isocyanate and a series of 9 nitrosoureas toward amino group of 4-amino-2,2,6,6 tetramethyl-piperidine-1-oxyl were examined with both the EPR method and the HPLC method which has been proposed by Brubaker et al. [Biochem. Pharmacol. 35:2359 (1986)]. Spin-labeled nitrosoureas we synthesized are included in this study since they have less toxicity or more efficiency than commercially available drug in some cases. The concentration of carbamoylated product was easily determined with the EPR spectra. There is a very high correlation (r = 0.982, t = 2.58, N = 10, p < 0.001) between the EPR and HPLC methods. Spin-labeled nitrosoureas showed lower carbamoylating activity than non-labeled analogues. The carbamoylating activity for these nitrosourea depended on the reactivity of isocyanate intermediate and almost independent of their half life. This rapid and simple EPR method is suitable for the detailed investigation of the rate and extent of carbamoylation reaction. PMID- 9350425 TI - Reference values for alpha-tocopherol and beta-carotene in the Whitehall II Study. AB - Plasma alpha-tocopherol, beta-carotene, serum lipids and their derived ratios were determined in British Civil Servants (n = 7177) at the second medical examination of the Whitehall II Study, a longitudinal study of cardiovascular disease. For plasma alpha-tocopherol the non-parametric 95% reference interval (90% confidence limits) for the total population was: 11.1 (10.9-11.3)-51.5 (50.6 52.7) mumol/l. For plasma beta-carotene the non-parametric reference interval for the total population was: 0.05 (0.05-0.05)-2.14 (2.08-2.21) mumol/l. The latter interval was wider than those previously published with a higher mean (0.61 mumol/l) and median (0.75 mumol/l). Plasma beta-carotene concentrations were higher in women than men with age-adjusted means of 0.70 and 0.57 mumol/l respectively (p < 0.001). This may reflect differences in diet, lifestyle and metabolism between the sexes. The alpha-tocopherol/cholesterol ratio, as in other surveys, did not vary with age. Among men, current- and ex-smokers had a higher alpha-tocopherol/cholesterol ratio than never-smokers with age-adjusted means of 4.18, 4.19 mumol/mmol and 4.05 mumol/mmol respectively. This difference is as yet unexplained. Follow-up of these subjects will help to clarify the role of antioxidant nutrients as protective factors for cardiovascular disease and cancer. PMID- 9350426 TI - Evaluation of antioxidant effectiveness of a few herbal plants. AB - We have screened a number of plants from the Indian soil for potential antioxidant properties out of which fifteen extracts were found to be positive. Leaves/bulk from the plants were crushed and extracted with organic solvents by three different ways. The first group of plants were extracted with CHCL3:CH3OH (2:1), evaporated, partitioned between petroleum ether and methanol (9:1), aqueous methanolic part re-partitioned between methanol:H2O (4:1) and dichloromethane. Methanol was evaporated from the aqueous methanolic part and extracted with n-butanol. The second group of plants were extracted with methanol followed by partitioning between petroleum ether and CH3OH. The rest of the extraction procedure was the same as above. A third extraction procedure was used for Ocimum sanctum which after extraction with CHCL3:CH3OH (2:1), partitioned between CCL4 and CH3OH:H2O (9:1). Aqueous methanolic part was repartitioned between CH3OH:H2O (4:1) and CHCl3 and CHCl3 soluble part was used for the study. Free radical scavenging activities of the plant extracts were examined by chemiluminescence method. Peroxyl radical was generated from 2,2'-azobis(2 amidinopropane) dihydrochloride (AAPH), superoxide radical (O2-) from xanthine/xanthine oxidase (XO) and hydroxyl radical (OH) from Xanthine/XO/FeCl3/ EDTA. In addition, O2- and OH. scavenging activities were also determined by cytochrome C reduction and deoxyribose oxidation methods, respectively. The results of this study demonstrate that these plant extracts possess potent antioxidant activities. PMID- 9350427 TI - Real time detection of reactions between radicals of lycopene and tocopherol homologues. AB - Laser flash photolysis of lycopene in homogeneous chloroform solution together with tocopherol homologues results in rapid formation of the lycopene radical cation and slower formation of tocopheroxyl radicals. Time-resolved detection by absorption spectroscopy of decay of the lycopene radical cation, of formation of the tocopheroxyl radicals, and of bleaching of lycopene has shown that alpha tocopherol is able to reduce the lycopene radical cation and thereby partially regenerate lycopene on a ms timescale. In contrast, lycopene is able to reduce the delta-tocopheroxyl radical, whereas an equilibrium exists between the lycopene radical cation and beta- or gamma-tocopherol. The relative stability of these antioxidant radicals is hence: alpha-tocopheroxyl > lycopene radical cation approximately beta-tocopheroxyl approximately gamma-tocopheroxyl > delta tocopheroxyl. PMID- 9350428 TI - Hydroxylation of salicylate and phenylalanine as assays for hydroxyl radicals: a cautionary note visited for the third time. AB - Hydroxylation of salicylate to 2,3- and 2,5-dihydroxy-benzoates (DHBs) is widely used as an index of hydroxyl radical (OH.) formation in vivo and in vitro. Several recent studies indicate that peroxynitrite can lead to generation of DHBs from salicylate and it is uncertain as to whether or not OH. is involved. A similar problem may occur in the use of phenylalanine as an OH. detector. Hence formation of hydroxylation products from salicylate (or phenylalanine) may not in itself be a definitive index of OH. generation, especially in cases where such generation in physiological systems is decreased by inhibitors of nitric oxide synthase. Determination of salicylate (or phenylalanine) nitration products can allow distinction between peroxynitrite-dependent aromatic hydroxylation and that involving "real" OH.. PMID- 9350430 TI - Malondialdehyde and 4-hydroxy-2-nonenal in plant tissue cultures: LC-MS determination of 2,4-dinitrophenylhydrazone derivatives. AB - The cytologically active secondary lipid peroxidation products, malondialdehyde (MDA) and 4-hydroxy-2-nonenal (HNE) have been detected as their 2,4 dinitrophenylhydrazone (DNP) derivatives in plant tissue cultures using LC-MS. This paper reports, for the first time, the use of LC-MS methodology to definitively identify 4-hydroxy-2-nonenal in plants. Limits of detection for the two derivatives are approximately 5 pmol (1.2 x 10(-9) g; 1 microM) and 0.1 pmol (3 x 10(-11) g; 20 nM) respectively. Mass spectrometer response was linear in the range from 2-200 microM DNP-MDA and 0.02-10 microM DNP-HNE. This methodology has been used to assess the formation of aldehydic secondary lipid peroxidation products in dedifferentiated callus cultures of Daucus carota. The finding that profiles of MDA and HNE can be correlated with embryogenic competence is of considerable interest as oxidative status has already been implicated as a regulatory factor in animal development. PMID- 9350429 TI - Oxidation of 2',7'-dichlorofluorescin by peroxynitrite. AB - The simultaneous production of nitric oxide and superoxide anion leads to the formation of peroxynitrite, a potent oxidant which may be an important mediator of cellular injury. Oxidation of dichlorofluorescin to the fluorescent dichlorofluorescein has been used as a marker for cellular oxidant production. The mechanisms of peroxynitrite-mediated oxidation of dichlorofluorescin to dichlorofluorescein were investigated. Chemically synthesized peroxynitrite (50 500 nM) induced the oxidation of dichlorofluorescin to dichlorofluorescein in a linear fashion. In addition, the simultaneous generation of nitric oxide and superoxide anion induced the oxidation of dichlorofluorescin to dichlorofluorescein, while nitric oxide (1-10 microM) alone under aerobic conditions did not. Peroxynitrite-mediated oxidation of dichlorofluorescin was not inhibited by the hydroxyl radical scavengers mannitol (100 mM) or dimethylsulfoxide (100 mM). Moreover, peroxynitrite-mediated oxidation of dichlorofluorescin was not dependent upon metal ion-catalyzed reactions. Furthermore, dichlorofluorescein formation was diminished at alkaline pH. These findings suggest that peroxynitrite-mediated dichlorofluorescein formation results directly from the protonation of peroxynitrite to form the conjugate peroxynitrous acid. L-cysteine was an efficient inhibitor (KI approximately 25 microM) of dichlorofluorescin oxidation through competitive oxidation of free sulfhydryls. Urate was a less efficient with a maximum inhibition of only 49%. These results demonstrate that dichlorofluorescin is efficiently oxidized by peroxynitrite. Therefore, under conditions where nitric oxide and superoxide are produced simultaneously, oxidation of dichlorofluorescin may be mediated by the formation of peroxynitrite. PMID- 9350431 TI - Oxidative modification of glutamine synthetase by amyloid beta peptide. AB - beta-Amyloid peptide (A beta), the main constituent of senile plaques and diffuse amyloid deposits in Alzheimer's diseased brain, was shown to initiate the development of oxidative stress in neuronal cell cultures. Toxic lots of A beta form free radical species in aqueous solution. It was proposed that A beta derived free radicals can directly damage cell proteins via oxidative modification. Recently we reported that synthetic A beta can interact with glutamine synthetase (GS) and induce inactivation of this enzyme. In the present study we present the evidence that toxic A beta(25-35) induces the oxidation of pure GS in vitro. It was found that inactivation of GS by A beta, as well as the oxidation of GS by metal-catalyzed oxidation system, is accompanied by an increase of protein carbonyl content. As it was reported previously by our laboratory, radicalization of A beta is not iron or peroxide-dependent. Our present observations consistently show that toxic A beta does not need iron or peroxide to oxidize GS. However, treatment of GS with the peptide, iron and peroxide together significantly stimulates the protein carbonyl formation. Here we report also that A beta(25-35) induces carbonyl formation in BSA. Our results demonstrate that beta-peptide, as well as other free radical generators, induces carbonyl formation when brought into contact with different proteins. PMID- 9350432 TI - The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase. AB - The tetrazolium dyes MTS and XTT were reduced to their soluble formazans by superoxide radical anions (O2-) produced by the oxidation of xanthine by xanthine oxidase under standard conditions. These reactions were compared to the well known reductions of NBT and cytochrome c by the xanthine/xanthine oxidase system. Reduction of the dyes was completely inhibited by superoxide dismutase (SOD). Rate constants for the reaction of MTS and XTT with O2- were estimated at 1.3 +/- .1 x 10(5) M-1S-1 and 8.6 x 10(4) M-1S-1 respectively. The stable MTS and XTT formazans have high extinction coefficients in the visible range which enable sensitive detection and quantification of superoxide radicals, avoiding some of the problems inherent in assays based on production of the insoluble NBT formazan. MTS and XTT have considerable potential both for the quantitative assay of radical production in living tissues and for the assay of superoxide dismutase activity in tissue extracts. Implications for the interpretation of cell culture growth assays which employ these dyes are discussed. PMID- 9350433 TI - Effect of aluminium ions on liposomal membranes as detected by Laurdan fluorescence. AB - We report here an investigation of the influence of aluminium on iron-induced peroxidation in brain model membranes. Laurdan fluorescence emission spectra and generalised polarisation measurements have been used to investigate how ferrous and aluminium ions can affect the phase components of phospholipid membranes. An increase in the generalised polarisation of oxidised liposomes with respect to controls has been observed, which reveals the presence of a less polar environment surrounding the probe that changes the properties of the bilayer. Aluminium has been shown to facilitate iron-mediated oxidation as detected from emission fluorescence spectra. However, no quantitative influence has been calculated relative to general polarisation and derived phase state determinations. The structural influence of aluminium on membranes may therefore be less significantly marked than initially expected. PMID- 9350434 TI - Involvement of different transduction pathways in NF-kappa B activation by several inducers. AB - Double-stimulation was used to demonstrate that, in a T lymphocytic cell line (CEM), phorbol myristate acetate (PMA) rapidly induced NF-kappa B through a signaling pathway which did not involve reactive oxygen species (ROS) and was different from the activation triggered by either H2O2 or tumor necrosis factor alpha (TNF-alpha). Since these latter compounds were known to activate NF-kappa B translocation in a redox-sensitive way, we have demonstrated that NF-kappa B activation by PMA was resistant to antioxidant N-acetyl-L-cysteine (NAC) and sensitive to kinase inhibitors staurosporine and H7 while activation by H2O2 or TNF-alpha were not. PMID- 9350435 TI - Effect of active oxygen radicals on protein and carbohydrate moieties of recombinant human erythropoietin. AB - Our previous study showed that active oxygen radicals generated from a Fenton system and a xanthine plus xanthine oxidase system caused serious loss of in vivo bioactivity of recombinant human erythropoietin (EPO), a highly glycosylated protein. In the present study, we characterized the oxidative modifications to the protein and carbohydrate moiety of EPO, which lead to a reduction of its bioactivity. In vitro bioactivity was reduced when EPO was treated with oxygen radicals generated from a Fenton system in the presence of 0.016 mM H2O2, and the reduction was directly proportional to the loss of in vivo bioactivity. SDS-PAGE analysis showed that dimer formation and degradation was observed under more severe conditions (Fenton reaction with 0.16 mM H2O2). The tryptophan destruction was detected at 0.016 mM H2O2 and well correlated with the loss of in vitro bioactivity, whereas loss of other amino acids were occurred under more severe conditions. Treatment with the Fenton system did not result in any specific damage on the carbohydrate moiety of EPO, except a reduction of sialic acid content under severe condition. These results suggest that active oxygen radicals mainly react with the protein moiety rather than the carbohydrate moiety of EPO. Destruction of tryptophan residues is the most sensitive marker of oxidative damage to EPO, suggesting the importance of tryptophan in the active EPO structure. Deglycosylation of EPO caused an increased of susceptibility to oxygen radicals compared to intact EPO. The role of oligosaccharides in EPO may be to protect the protein structure from active oxygen radicals. PMID- 9350436 TI - Molecular mechanisms of apoptosis in HL-60 cells induced by a nitric oxide releasing compound. AB - Nitric oxide (NO) generated from 1-hydroxy-2-oxo-3, 3-bis(2-aminoethyl)-1 triazene (NOC 18), an NO-releasing compound, induced monocytic differentiation of human promyelocytic leukemia HL-60 cells as assessed by expression of nonspecific esterases and morphologic maturation. Simultaneously, DNA fragmentation and morphological alterations typical of apoptosis were also induced. To investigate the mechanisms of apoptosis during differentiation of HL-60 cells induced by NO, the endogenous levels of Bcl-2 and Bax were assessed by immunoblotting. Treatment of cells with NOC 18 slightly reduced the level of Bcl-2 followed by Bax. These changes might be involved in the induction of apoptosis. The involvement of the activation of the interleukin-1 beta converting enzyme (ICE) family of proteases (caspases), such as ICE and CPP32, in the pathways was also investigated. CPP32, but not ICE, was strongly activated in response to NOC 18 stimulation, thereby implicating CPP32-like activity in the induction of apoptosis. Moreover, the possible involvement of tyrosine phosphorylation in apoptosis was investigated. Pretreatment of cells with herbimycin A, an inhibitor of tyrosine kinases, suppressed DNA fragmentation and CPP32-like activity, whereas pretreatment with vanadate, an inhibitor of tyrosine phosphatases, enhanced both parameters, suggesting that tyrosine phosphorylation might be involved in the pathways of apoptosis in HL-60 cells induced by NO. PMID- 9350437 TI - The effect of the phenolic antioxidant ferulic acid on the oxidation of low density lipoprotein depends on the pro-oxidant used. AB - The action of ferulic acid during the oxidation of LDL has been investigated using both copper ions and the haem protein metmyoglobin as pro-oxidants. The results demonstrate the ability of ferulic acid to act as a pro-oxidant when LDL oxidation is induced by copper at concentrations of the phenolic acid which are protective when the LDL oxidation is mediated by metmyoglobin. The suggested mechanism involves the reduction of Cu2+ to Cu+ by ferulic acid resulting in the production of the ferulic phenoxyl radical. PMID- 9350438 TI - Leptin: the voice of the adipose tissue. AB - Leptin is a newly discovered hormone that acts as a feedback signal from the adipose tissue. It plays a pivotal role in the modulation of neuronal and hormonal systems involved in the regulation of body weight and reproductive functions. This brief overview focuses on the regulation of circulating leptin levels and leptin in extreme clinical states of body weight, summarizing mainly results from the University of Giessen in collaboration with other groups. Finally, a possible role for leptin is presented. PMID- 9350439 TI - Functional role of insulin-like growth factor binding proteins. AB - Insulin-like growth factors (IGF-I, IGF-II) are important regulators of cell division and differentiation. In their free form, IGFs form a complex with specific binding proteins (IGFBPs), six of which have now been characterized. These IGFBPs differ in their ability to bind IGF-I and/or IGF-II, and, depending on their location and metabolic circumstances, they inhibit or augment IGF action to varying extents. New findings have changed our understanding of IGFBPs, which are now considered to be major regulators of IGF action and IGF transport proteins between compartments. Recently, the IGF-independent action of IGFBPs was discovered. Levels of IGFBP-3, for instance, are important indicators of states of altered growth hormone secretion and action, as well as being important during treatment with growth hormone or IGF-I. IGFBP-1 and -2 levels reflect changes related to nutrition, insulin secretion, foetal development and malignant state. The unravelling of the intricate interactions of IGFs, IGFBPs and their targets is bound to influence our understanding of the physiology and development of biological systems. Likewise, the possibility of regulating the IGFBP system is likely to open up new fields in the treatment of diseases in the future. PMID- 9350440 TI - Disturbances in the growth hormone-insulin-like growth factor axis in children and adolescents with different eating disorders. AB - Numerous endocrine abnormalities of the growth hormone (GH)-insulin-like growth factor axis have been described in patients with both anorexia nervosa and obesity during childhood and adolescence. These alterations include changes in the levels of 24-hour spontaneous GH secretion, high-affinity, low-capacity GH binding protein (GHBP), IGF-I, IGF-II and the IGF binding proteins (IGFBPs). However, the existing information is sometimes confusing and contradictory. Furthermore, little or no data in these pathologies are available concerning IGFBP-2 or free IGF-I. We have analysed the GH-IGF axis in large populations of adolescents with anorexia nervosa and prepubertal children with exogenous obesity. These patients were studied at the time of diagnosis and at two timepoints during nutritional therapy and normal weight recovery. The results of these studies using age- and sex-matched controls are described here. PMID- 9350441 TI - Mechanisms of growth failure in non-growth-hormone deficient children of short stature. AB - So far, the clinical evaluation of short children has focused on the measurement of immunoreactive growth hormone (GH) in the blood to determine if the growth retardation is due to GH deficiency. However, GH-dependent short stature may be caused by defects in either the secretion of bioactive GH or by the inability to respond to GH. Both GH secretion and GH responsiveness should, therefore, be evaluated when investigating the cause of short stature. Recent advances in molecular biology have generated new ways of studying different molecules involved in growth regulation, and new genetic defects have been identified in short children. PMID- 9350442 TI - Growth hormone treatment of children with short stature secondary to intra uterine growth retardation: effect of 2 years' treatment and 2 years' follow-up. AB - Growth hormone (GH) treatment has been proposed to improve final height in patients with short stature associated with intra-uterine growth retardation (IUGR). In this study, 30 prepubertal patients aged 9.5 +/- 0.9 years with IUGR and normal GH secretion on pharmacological testing were treated with GH. These patients had a mean birth length of -3.11 +/- 0.80 SDS, and mean growth retardation of -2.58 +/- 0.49 SDS for chronological age. GH, 1.4 IU/kg/week (= 0.07 mg/kg/day), was given for 2 years. Height gain (calculated as the difference of height SDS at baseline and after 2 years) was 1.3 +/- 0.4 SD and was not significantly correlated with height SDS or growth velocity at baseline. These data confirm that 2 years of recombinant human GH treatment increases height gain in patients with IUGR. Two years after treatment interruption, mean gain was maintained at +1.08 SDS and 83% of the children had normal height. PMID- 9350443 TI - Growth hormone therapy and malignancy. AB - The possibility that human growth hormone (GH) replacement therapy might either increase the risk of cancer recurrence in a child who has previously been treated for a brain tumour or leukaemia, or induce de novo cancer, has worried paediatricians for a number of years. Concern arises from animal experiments, the association of acromegaly with malignancy, and the Japanese experience of a cluster of de novo leukaemia cases in children treated with GH. It is reassuring that so far the results from single centre studies and from the pharmaceutical industry surveillance programmes have shown no evidence of an increased risk of malignancy, recurrent or de novo. The confidence intervals, however, are wide and the scientific nature of these studies is flawed as there has never been a prospective randomized study of GH replacement in children with radiation-induced GH deficiency. For clinical reasons, such a study is unlikely to be performed and therefore surveillance must be maintained at a very high level. PMID- 9350444 TI - Gonadotrophin and thyrotrophin receptors. AB - Gonadotrophin and thyrotrophin receptors belong to a subgroup of G-protein coupled receptors. These receptors are characterized by a large extracellular domain that is responsible for the binding of the hormone. Soluble receptors, such as some luteinizing hormone receptors, arise from premessenger RNA alternative splicing, or, in the case of thyroid-stimulating hormone (TSH) receptors, by the cleavage and shedding of the ectodomain. Follicle-stimulating hormone and TSH receptors are restricted to the basolateral domain of their target cells. These receptors are also present in endothelial cells of target organ vessels and are involved in hormone transcytosis. Various genetic abnormalities of these receptors have been described. PMID- 9350445 TI - Effect of growth hormone on cardiac function. AB - At present, there is a growing body of evidence implicating growth hormone (GH) and/or insulin-like growth factor-I (IGF-I) in the intricate cascade of events connected with the regulation of heart development and hypertrophy. In addition, advanced clinical manifestations of abnormal GH levels almost always include impaired cardiac function, which may reduce life expectancy. This finding is related both to a primary impairment of heart structure and function and to metabolic changes such as hyperlipidaemia, increased body fat and premature atherosclerosis. Acromegalic cardiomyopathy is better correlated with disease duration than with GH or IGF-I levels. Myocardial hypertrophy with interstitial fibrosis, lymphomononuclear infiltration and areas of monocyte necrosis often result in increased right and left ventricular mass concentric hypertrophy. Conversely, patients with childhood or adult-onset GH deficiency (GHD) have a reduced left ventricular mass (LVM) and ejection fraction (EF) and the indices of left ventricular systolic function remained markedly depressed during exercise. Cardiac function is reported to improve during octreotide and GH replacement treatment in acromegaly and GHD, respectively. The evidence that GH can increase cardiac mass suggests its use in the treatment of idiopathic dilated cardiomyopathy. In a recent study on such patients, the administration of recombinant GH (rGH) was demonstrated to increase myocardial mass and reduce the size of the left ventricular chamber, resulting in improved haemodynamics, myocardial energy metabolism and clinical status. PMID- 9350446 TI - Resistance to thyroid hormone. AB - Resistance to thyroid hormone (RTH) is usually dominantly inherited and is characterized by elevated free thyroid hormones in the serum and failure to suppress pituitary thyroid stimulating hormone (TSH) secretion with variable refractoriness to hormone action in peripheral tissues. Two major forms of the disorder are recognized: asymptomatic individuals with generalized resistance (GRTH) and patients with thyrotoxic features, suggesting predominant pituitary resistance (PRTH). Molecular genetic analyses indicate that both GRTH and PRTH are associated with diverse mutations in the thyroid hormone receptor beta gene, which localize to three regions in the hormone binding domain of the receptor. In addition to being functionally impaired, the mutant receptors are also able to inhibit their wild-type counterparts in a dominant negative manner. Recognized features of RTH include failure to thrive, growth retardation and attention deficit hyperactivity disorder in childhood, and goitre and thyrotoxic cardiac symptoms in adults. The pathogenesis of variable tissue resistance is not fully understood but may be related to the differing tissue distributions of a and b thyroid hormone receptors and variable dominant negative activity of mutant receptors on different target genes. PMID- 9350447 TI - New pathophysiological mechanisms for hyperthyroidism. AB - From the analysis of accumulated data about adrenergic receptors, it was hypothesized that some mutations within the thyroid-stimulating hormone (TSH) receptor would activate it constitutively. As TSH positively controls the function, differentiation and growth of thyrocytes by cyclic AMP-dependent mechanisms, such somatic or germline mutations would cause readily identifiable phenotypes in the form of monoclonal hyperfunctional benign tumours or hereditary dominant hyperthyroidism, respectively. This hypothesis turned out to be correct: of 29 hyperfunctioning adenomas, 23 were found to have a mutation at various positions of the serpentine portion of the TSH receptor. In five families with autosomal dominant hyperthyroidism and one sporadic case, five additional different mutations were identified. Altogether, 20 different residues have been found mutated in toxic adenomas or toxic thyroid hyperplasia. It is tempting to speculate that as the TSH receptor is already 'noisy' in its wild-type state, it would be more readily activated by mutations than others. PMID- 9350448 TI - New aspects of thyroid immunity. AB - Like all autoimmune diseases, those affecting the thyroid arise as the result of an interaction between genetic, environmental and endogenous factors. Genes outside the human leukocyte antigen (HLA) complex play a critical role, with recent associations described between a T cell surface receptor for the costimulatory signal B7 (provided by antigen presenting cells; CTLA-4), and interleukin-1 (IL-1) receptor antagonist polymorphisms and autoimmune thyroid disease. Birth weight and hormonal changes constitute important endogenous influences on susceptibility, while iodine intake, stress, infections and toxins are all environmental factors. Once initiated, events within the thyroid determine whether or not autoimmunity is maintained. Originally, HLA class II molecule expression by thyroid cells was thought to be a likely trigger in thyroid autoimmunity. However, thyroid cells cannot provide a necessary costimulatory signal, required in addition to class II molecule-mediated antigen presentation, making them incapable of initiating activation of autoreactive T cells. Moreover, class II-positive thyroid cells can induce peripheral tolerance in T cells and so class II expression can be viewed as a protective mechanism. Later, in established disease, costimulatory signals are not required by T cells for further stimulation, and class II expression could then exacerbate disease. PMID- 9350449 TI - Genetics of type 1 insulin-dependent diabetes mellitus. AB - Many human insulin-dependent (type 1) diabetes mellitus (IDDM) genes have recently been mapped. The HLA region on chromosome 6 and the insulin gene promoter on chromosome 11p have been extensively investigated. Genome scanning of many families with several affected individuals has successfully mapped other predisposing loci situated on different chomosomal regions. In addition, the positional cloning of IDDM susceptibility genes is in progress. Advances in high throughput genotyping and large-scale sequencing, and the availability of high density maps of human genes will help to identify IDDM genes. Understanding the genetic basis of IDDM will aid in the design of new strategies to prevent or cure the disease. PMID- 9350450 TI - Immune mechanisms leading to type 1 insulin-dependent diabetes mellitus. AB - Insulin-dependent diabetes mellitus (IDDM) results from the autoimmune destruction of the endocrine (beta) cells responsible for the secretion of insulin. Experimental and human studies have resulted in unpredicted developments over the past ten years. Against most predictions, a long list of possible autoantigens has been defined, most of which are not beta-cell specific. Seemingly, up to 10 to 20 genetic regions, rather than 2 to 3 as initially predicted, carry markers that associate them with IDDM. Finally, there is accumulating evidence that beta cells are not a passive bystander in the event that leads to the activation of the diabetes autoimmune reaction. Whether the immune system or the islet, or both, are at the initiation of the disease process is a challenge for the near future. PMID- 9350451 TI - Sardinia: a battlefield approach to type I diabetes epidemiology. Sardinia-IDDM Study Groups. AB - Sardinia and Finland have the highest incidence of insulin-dependent diabetes mellitus (IDDM) in the world. Therefore, both regions represent ideal observatories for investigating the environmental, genetic and immunological factors which have led to this dramatic increase. We have concentrated our efforts on Sardinia. Among several projects, there is the mapping of the island for hot and cold spots for overt IDDM. In order to map the island for pre-IDDM, we have collected and bled around 10,000 school children (age 6-14 years) and we are now in the process of enrolling around 30,000 new-born babies. We report here our initial results, which show that progression to IDDM is accompanied in both cohorts by the presence of a combination of islet-cell antibodies with either glutamic acid decarboxylase or IA-2 antibodies or both. This approach should lead to the design of reliable models of IDDM prediction in the general population, which will benefit an early insulin treatment and, hopefully, an effective prevention of the disease. PMID- 9350452 TI - Insulin in diabetes prevention. AB - Insulin-dependent diabetes (IDD) is a chronic immune-endocrine disease in which there is a progressive destruction of insulin-secreting pancreatic beta cells, caused primarily by autoreactive T cells. Many islet cell proteins including insulin, glutamic acid decarboxylase, and tyrosine phosphatase antigens (IA-2) are targeted by the autoimmune responses in IDD patients. Since its discovery 75 years ago, insulin has been the major player in the clinical management of hyperglycaemia in these patients. The morbidity and mortality associated with IDD derives mainly from the complications of the disease. However, routine insulin injections seldom achieve a consistent, near-normal glucose level, where multiple daily doses of the hormone involve considerable restrictions to a normal lifestyle. In terms of economics, the management of diabetes is expensive, and in the USA diabetes alone accounts for one seventh of the healthcare budget. These clinical, lifestyle and economic issues emphasize the need to investigate alternative preventative measures in IDD treatment. Recent reports suggest a pivotal role for insulin in various aspects of the immune system. In this study, insulin and B-chain were used to modulate autoimmune responses in non-obese diabetic mice, findings which have therapeutic implications in man. PMID- 9350454 TI - Nosocomial outbreak of scabies clinically resistant to lindane. PMID- 9350455 TI - Does a cheaper mask save money? The cost of implementing a respiratory personal protective equipment program. PMID- 9350453 TI - Management of insulin-dependent diabetes mellitus in adolescents. AB - There are many reasons for a specific management plan for adolescents with insulin-dependent diabetes mellitus (IDDM). Although most new patients can be managed initially on an outpatient basis, as they reach their teenage years, blood glucose control deteriorates and insulin requirements increase. Eating disorders, missing insulin injections, excess of alcohol all contribute to poor glycaemic control. Teenagers who have had diabetes for a number of years may have developed microvascular complications and disturbances of growth can occur, more often in girls than in boys. The transition to adult care continues to be a significant problem both for paediatricians and patients. Diabetes management during adolescence requires a team effort involving nurse educator, dietitian, paediatric diabetologist and, possibly, a social worker, with referral to specialists if necessary. Organizing this type of care requires a Regional Paediatric Diabetes Centre. PMID- 9350456 TI - Susceptibility of vancomycin-resistant enterococci to environmental disinfectants. PMID- 9350457 TI - Natural history of colonization with vancomycin-resistant Enterococcus faecium. PMID- 9350458 TI - Hepatitis B immunization of hospital employees in an endemic area: should we screen? PMID- 9350459 TI - Selection and use of vaccines for healthcare workers. PMID- 9350460 TI - Cost-effectiveness of hepatitis A vaccination in healthcare workers. AB - OBJECTIVE: To study the cost-effectiveness of vaccination for hepatitis A. SETTING: Hypothetical analysis of students currently enrolled in medical school in the United States. METHOD: A Markov-based model was developed using data from the literature, actual hospital costs, and an annual discount rate of 5%. The incidence rate was based on the lowest annual rate for the US population during the past decade. RESULTS: Over the lifetimes of students currently in medical school, the model estimated that there would be 286 hepatitis A cases with four deaths and 107 lost years of life. With routine vaccination, these numbers would decrease to 17, 0.3, and 6, respectively. The costs per life-year saved and quality adjusted life-year saved were $58,000 and $47,000, respectively. Serologic screening prior to vaccination was less cost-effective than universal vaccination. If the incidence of hepatitis A was underestimated by a factor of 5, the cost per life-year saved would decrease to $5,500. If the incidence of hepatitis was underestimated by a factor of 10, vaccination would result in a net cost savings. CONCLUSION: We conclude that the cost per life-year saved by routine hepatitis A vaccination was similar to many other standard medical modalities. For routine vaccination of medical students to be cost-saving, the incidence rate for hepatitis A must be at least 10 times higher than the rate presently reported for the general population. Serological screening prior to vaccination was not cost-effective. PMID- 9350461 TI - Noncompliance with universal precautions and the associated risk of mucocutaneous blood exposure among Danish physicians. AB - OBJECTIVE: To study the compliance, and reasons for noncompliance, with Universal Precautions and the associated circumstances of mucocutaneous blood exposure (MCE) among Danish physicians. DESIGN: A nationwide questionnaire survey. SETTING: All Danish hospitals. PARTICIPANTS: All hospital-employed physicians. RESULTS: Of 9,384 questionnaires, 6,256 (67%) were returned, and 6,005 were eligible for analysis. Only 35% of respondents were compliant with the basic principle of Universal Precautions. Compliance with specific barriers in the preceding week among "surgeons and pathologists" and "other physicians" was as follows: gloves, 63.0% and 23.4%; masks, 55.2% and 17.6%; and protective eyewear, 11.5% and 4.0%, respectively. Common arguments for non-compliance were "interferes with working skills," "forget," "wear spectacles," "not available," "too much trouble to get," or "gloves do not fit." Detailed descriptions of 741 MCEs were obtained. Blood splashes in the eyes (n = 320) was the most common MCE in surgical specialties and pathology, whereas blood on the hands (n = 290) was most common in other specialties. In 20% of MCEs of the eyes, the exposure occurred despite the use of spectacles. An estimated 84% to 98% of MCEs potentially would have been preventable had appropriate barriers been worn. More than one half of MCEs were preventable by two interventions only: compulsory use of protective eyewear during operations and use of gloves during insertion of peripheral intravenous catheters. CONCLUSION: Compliance with Universal Precautions is unacceptably low, as reflected by the circumstances of MCE. Increased efforts to ensure education in Universal Precautions, easy accessibility of protective barriers, and improved design of the barriers are necessary to improve compliance and reduce the risk of MCE. PMID- 9350462 TI - The infection control practices of general dental practitioners. AB - OBJECTIVES: To investigate the infection control practices of general dentists in Ontario in 1994. DESIGN: Confidential coded questionnaires were mailed to all general dental practitioners in Ontario (n = 5,176), with three follow-up attempts. Data were analyzed using Pearson's chi-squared test and multiple logistic regression. SETTING: Offices of general dental practitioners in Ontario. PARTICIPANTS: General dental practitioners actively involved in treating patients. RESULTS: The response rate adjusted for nondelivery was 70%. A high proportion of respondents reported using gloves (always, 91.8%; sometimes, 7.8%), masks (always, 74.8%; sometimes, 21.1%), or protective eyewear (always, 83.6%; sometimes, 13%); heat sterilization of handpieces (83.9%); and hepatitis B (HBV) vaccination of dentists (92.3%). However, only 61.4% of respondents reported HBV vaccination of all clinical staff, and 87.7% used additional precautions for patients with human immunodeficiency virus (HIV). Significant predictors of the use of recommended infection control procedures (i.e., always using gloves, masks, and eye protection; heat sterilization of handpieces; HBV vaccination for dentist and staff; and no extra precautions for patients with HIV) were age < 40 years (odds ratio [OR], 2.6), lack of concern regarding increased personal risk (OR, 2.0) or costs of infection control procedures (OR, 1.5), and knowledge of the low infectivity of HIV after a needlestick injury (OR, 2.0) and that infection control procedures for HBV are adequate for HIV (OR, 2.7). CONCLUSION: Additional education is required to promote a more realistic perception of risk of HIV transmission in the dental office and the use of all recommended infection control practices, including Universal Precautions. PMID- 9350463 TI - Serratia marcescens outbreak associated with extrinsic contamination of 1% chlorxylenol soap. AB - OBJECTIVES: To determine risk factors for Serratia marcescens infection or colonization, and to identify the source of the pathogen and factors facilitating its persistence in a neonatal intensive-care unit (NICU) during an outbreak. DESIGN: Retrospective case-control study; review of NICU infection control policies, soap use, and handwashing practices among healthcare workers (HCWs); and selected environmental cultures. SETTING: A university-affiliated tertiary care hospital NICU. PATIENTS: All NICU infants with at least one positive culture for S marcescens during August 1994 to October 1995. Infants who did not develop S marcescens infection or colonization were selected randomly as controls. RESULTS: Thirty-two patients met the case definition. On multivariate analysis, independent risk factors for S marcescens infection or colonization were having very low birth weight (< 1,500 g), a patent ductus arteriosus, a mother with chorioamnionitis, or exposure to a single HCW. During January to July 1995, NICU HCWs carried their own bottles of 1% chlorxylenol soap, which often were left standing inverted in the NICU sink and work areas. Cultures of 16 (31%) of 52 samples of soap and 1 (8%) of 13 sinks yielded S marcescens. The 16 samples of soap all came from opened 4-oz bottles carried by HCWs. DNA banding patterns of case infant, HCW soap bottle, and sink isolates were identical. CONCLUSIONS: Extrinsically contaminated soap contributed to an outbreak of S marcescens infection. Very-low-birth-weight infants with multiple invasive procedures and exposures to certain HCWs were at greatest risk of S marcescens infection or colonization. PMID- 9350465 TI - Effect of a comprehensive program to reduce needlestick injuries. AB - The Arlington Hospital Needlestick Injury (NSI) Prevention Program was created to protect healthcare workers from NSI and to assess the effectiveness of our interventions. Interventions included revising NSI policy and procedures. The average NSI rate dropped from 109 to 43 per year after the interventions, over a period of 4 years. PMID- 9350464 TI - Healthcare workers' attitudes and compliance with universal precautions: gender, occupation, and specialty differences. AB - We describe variations in healthcare workers' attitudes toward double gloving and reporting needlesticks, and in their readiness to comply with double gloving and hepatitis B vaccine. Differences related to occupation, specialty, and gender have implications for the need to tailor interventions for specific groups of healthcare workers to improve compliance with Universal Precautions. PMID- 9350466 TI - Nosocomial acquisition of pertussis diagnosed by polymerase chain reaction. AB - Awareness is growing of the role of pertussis infection among adolescents and adults, and of the possibility of nosocomial transmission. However, pertussis remains difficult to diagnose. We report a case of nosocomially acquired pertussis in a healthcare worker, diagnosed by polymerase chain reaction. The high sensitivity and rapid turnaround time for this test is useful to confirm the diagnosis when advising a healthcare worker to refrain from working. PMID- 9350467 TI - Occupational blood exposures in dentistry: a decade in review. AB - This review summarizes data from self-reported and observational studies describing the nature, frequency, and circumstances of occupational blood exposures among US dental workers between 1986 and 1995. These studies suggest that, among US dentists, percutaneous injuries have declined steadily over the 10 year period. Data also suggest that, in 1995, most dental workers (dentists, hygienists assistants, and oral surgeons) experienced approximately three injuries per year. Work practices (eg, using an instrument instead of fingers to retract tissue), safer instrumentation or design (eg, self-sheathing needles, changes in dental-unit design), and continued worker education may reduce occupational blood exposures in dentistry further. PMID- 9350468 TI - Product evaluation. AB - As healthcare budgets shrink, infection control personnel must become more involved in the process by which their institutions choose products and equipment. This article suggests an approach that individuals or programs can use to assess products in a consistent and thorough manner. PMID- 9350469 TI - Biochemistry and histochemistry of oxidant stress and lipid peroxidation. AB - Oxidant stress--i.e. the prevalence within the cell of oxidizing species over the cellular antioxidant potential--is recognized as a primary factor in the pathogenesis of a series of important human pathologies. The possibility of a preventive or therapeutic intervention by means of antioxidant factors, including those naturally contained in diet, increases the importance of a correct understanding of the molecular mechanisms involved in such pathologies. A number of sophisticated biochemical methods are available, for the monitoring of nearly all oxidant stress-related processes; their applicability to in vivo conditions is however limited. In these conditions, the histochemical visualization in tissue sections and isolated cells of selected molecular markers for oxidative phenomena can often provide valuable information about the distribution of processes in vivo. This paper includes an overview of the basic biochemical reactions occurring during oxidant stress and lipid peroxidation, as well as of the main histochemical studies published in the field. PMID- 9350470 TI - Antioxidative vitamins in prevention of ischemia/reperfusion injury. AB - Involvement of oxygen free radicals in ischemia-/reperfusion injury is based on measurement of increased products of lipid peroxidation after organ ischemia and restoration of blood flow during surgical operations and reperfusion of organ transplants. In cardiology inverse epidemiological correlations between plasma vitamin E levels and mortality due to ischemic heart disease, as well as beneficial effects of vitamin E on experimentally induced oxidative damage to the heart support the hypothesis, that vitamin E might have a protective role against myocardial ischemia-/reperfusion injury. In abdominal surgery efficiency of free radical scavengers has been intensively studied on animal models of hepatic ischemia and reperfusion. Examination of free radical scavengers and adenosine metabolites in liver tissue during hepatic ischemia revealed that vitamin E and glutathione levels as well as hepatic adenosine triphosphate levels are decreased during hepatic ischemia and reperfusion. The beneficial effects of alpha tocopherol on hepatic viability and survival rate after ischemia and reperfusion demonstrated in these studies will be of great importance concerning further studies in organ preservation. In clinical kidney transplantation prevention of lipid peroxidation and improvement in kidney viability and function was demonstrated after infusion of a multivitamin cocktail in a prospective randomised study. PMID- 9350471 TI - Interaction of antioxidative micronutrients with host defense mechanisms. A critical review. AB - An adequate host defense activity critically depends upon the micronutrient status of an individual among which the cellular oxidant-antioxidant balance is an important determinant. Oxidative burst is part of the physiological function of phagocytes connected to a massive production and release of reactive oxygen intermediates. At the same time, maintenance of the functional capacity of the host defense system in fighting against microorganisms and foreign antigens is significantly affected by the various reactive oxygen species. In order to compensate for this critical condition phagocytes do show active uptake and physiologically high intracellular concentrations of antioxidants. Thus, optimal function of the host defense system depends upon an adequate supply of antioxidative micronutrients and, on the other hand, impaired host defense activity can act as a very early and sensitive marker of marginal deficiency of antioxidant micronutrients. Assessment of immune functions can serve as an important preventative diagnostic tool in the detection of marginal but functionally relevant micronutrient deficiencies. Intervention into the functionally deficient antioxidant micronutrient status can act as the appropriate preventative or therapeutic treatment with higher efficacy and less adverse effects than direct pharmacological modulation of single immune functions by specific mediators. PMID- 9350472 TI - Effect of antioxidative vitamins on immune function with clinical applications. AB - Infection and trauma cause inflammatory stress in patients. Tissue damage, enhanced inflammatory mediator production and suppressed lymphocyte function may occur as a consequence. The antioxidative vitamins, ascorbic acid and the tocopherols, are important not only for limiting tissue damage but also in preventing increased cytokine production which is a consequence of excessive activation of NF kappa B. Glutathione is a major endogenous antioxidant and is important for lymphocyte replication. Two vitamins, vitamin B6 and riboflavin participate in the maintainance of glutathione status. The former vitamin acts as a cofactor in the synthesis of cysteine (the rate limiting precursor for glutathione biosynthesis) and the latter vitamin is a cofactor for glutathione reductase. Deficiencies in tocopherol, vitamin B6 and riboflavin reduce cell numbers in lymphoid tissues of experimental animals and produce functional abnormalities in the cell mediated immune response. Ascorbic acid and tocopherols exert anti-inflammatory effects in studies in man and animals. In humans, dietary supplementation with ascorbic acid, tocopherols and vitamin B6 enhances a number of aspects of lymphocyte function. The effect is most apparent in the elderly. PMID- 9350473 TI - Antioxidative vitamins in prematurely and maturely born infants. AB - Postnatally a rapid change occurs from a relatively hypoxic to a relatively hyperoxic environment, especially during artificial ventilation with all risks of ROS-formation. Among the non enzymatic antioxidative strategies the vitamins E, C, A and B2 are of major importance. Vitamin E is considered the most important radical scavenging vitamin of the lipid soluble compartment. Hereby vitamin E itself is converted into a radical which is handed over to vitamin C and glutathione into the water soluble compartment. The vitamin E content of the fetus increases with the fetal fat mass mainly during the last trimester of pregnancy. Placenta is only slightly permeable to lipid soluble vitamins. Vitamin E deficiency may rapidly develop typically at about 6-8 weeks of age. Vitamin E is able to prolong significantly the onset of retinopathic changes during oxygen therapy and may prevent intraventricular hemorrhage. Vitamin C is together with glutathione a major representative of the non enzymatic antioxidative system in the water soluble compartment. The best determinant of the vitamin C status is its concentration in leukocytes. Vitamin C reduces iron to the divalent state which supports the hydroxyl radical formation (Haber-Weiss reaction). This should be considered mainly in cases of intraventricular hemorrhage. Vitamin B2 acts mainly as cofactor of glutathione reductase which keeps glutathione in the reduced state. It can therefore be considered an indirect antioxidative vitamin. Vitamin B2 is destroyed by light. Phototherapy has been recognized as a cause of riboflavin deficiency. Vitamin A comprises all retinols with properties like trans-retinol. Retinol storage in the fetal liver increases during late pregnancy. In both, premature and mature newborns, the serum concentrations amount to only about 50% of those of their mothers. Vitamin A has a paramount importance for fetal lung development, because the individual surfactant proteins are selectively regulated by retinoic acid. PMID- 9350474 TI - Vitamin C supplementation and the common cold--was Linus Pauling right or wrong? AB - In 1970 Linus Pauling claimed that vitamin C prevents and alleviates the episodes of the common cold. Pauling was correct in concluding from trials published up till then, that in general vitamin C does have biological effects on the common cold, but he was rather over-optimistic as regards the size of benefit. His quantitative conclusions were based on a single placebo-controlled trial on schoolchildren in a skiing camp in the Swiss Alps, in which a significant decrease in common cold incidence and duration in the group administered 1 g/day of vitamin C was found. As children in a skiing camp are not a representative sample of the general population, Pauling's extrapolation to the population at large was too bold, erring as to the magnitude of the effect. Nevertheless, Pauling's general conclusion that vitamin C has physiological effects on the common cold is of major importance as it conflicts with the prevailing consensus that the only physiological effect of vitamin C on human beings is to prevent scurvy. PMID- 9350475 TI - Oxidative stress and signal transduction. AB - Reactive oxygen intermediates (ROIs) are an evolutionarily ancient threat to all organisms. Both prokaryotic and higher eukaryotic cells are able to alter their genetic program in response to changes in the intracellular levels of ROIs. In bacteria and yeast, this response leads to the new synthesis of proteins that protect cells from the consequences of oxidative damage, such as DNA strand breaks, lipid peroxidation and oxidative damage of proteins. Intriguingly, higher organisms have also evolved cellular mechanisms to actively produce ROIs. There is increasing evidence that ROIs fulfil an important role as second messengers involved in signal transduction. We have proposed that ROIs have been conserved throughout evolution as universal pathogen messengers turning on genes with important functions in the immune response and cell proliferation. The higher eukaryotic transcription factors NF-kappa B and AP-1 will be described as proteins which are regulated by ROIs under a great variety of pathogenic conditions and initiate the new expression of genes with important roles in immune, inflammatory and other pathogen-related genetic responses. PMID- 9350477 TI - Management of osteoporosis: is there a role for vitamin K? AB - Vitamin K is required for the biological activity of several coagulation factors, which is considered as the classical function of vitamin K. Recent research, however, suggests a role of vitamin K in bone metabolism. The metabolic role of vitamin K is to facilitate the carboxylation of glutamyl to gamma-carboxyglutamyl residues. Besides the hepatic tissue, in which the clotting factors are produced gamma-carboxyglutamyl-containing proteins are also abundantly available in bone tissue. Osteocalcin accounts for up to 80% of the total gamma-carboxyglutamyl content of mature bone. Human carboxylated osteocalcin contains 3 gamma carboxyglutamyl residues which confer a highly specific affinity to the calcium ion of the hydroxyapatite molecule. Besides the gamma-carboxylation of osteocalcin vitamin K may also affect other parameters of bone metabolism, such as calcium hemostasis, and prostaglandin E2 and interleukin 6 production. Evidence from observational studies and first intervention trials indicate that vitamin K intakes much higher than the current recommendations improved biochemical markers of bone formation as well as bone density. In conclusion, the mechanistic data as well as the observational data and the results of the first controlled clinical trials in humans point to a beneficial effect of additional intakes of vitamin K in bone health. PMID- 9350476 TI - Signalling functions of alpha-tocopherol in smooth muscle cells. AB - alpha-Tocopherol but not beta-tocopherol, activates protein phosphatase 2A, decreases protein kinase C activity and attenuates smooth muscle cell proliferation at physiological concentrations. beta-Tocopherol prevents the effects of alpha-tocopherol. Inhibition of protein kinase C alpha, but not of the other isoforms, by the inhibitor Go6976 prevents the effect of alpha-tocopherol. Protein kinase C alpha, immunoprecipitated from alpha-tocopherol treated cells, is less phosphorylated and inactive. It is proposed that the specific activation of protein phosphatase 2A by alpha-tocopherol results in dephosphorylation and inactivation of protein kinase C alpha. Finally, this cascade of events leads to smooth muscle cell proliferation inhibition. PMID- 9350478 TI - Comparative assessment of the activity of beta-carotene, retinoyl-beta-D glucuronide and retinoic acid on growth and differentiation of a human promyelocytic leukemia cell line HL-60. AB - We compared the ability of all-transretinoic acid (RA), all-trans-retinoyl-beta-D glucuronide (RAGL), and all-trans-beta-carotene (BC) to inhibit growth and to induce differentiation of the human promyelocytic leukemia cell line HL-60 into morphologically mature granulocytes. BC was made water-soluble by the solutol solvent-system. RA (1 microM) could induce differentiation of 85% of the HL-60 cells after a total incubation time of 180 hours, RAGL (5 microM) induced 64% of the cells, whereas 33% of the HL-60 cells were differentiated after incubation with BC (10 microM), which was determined by assessing cell functional capacity to reduce nitroblue tetrazolium dye in response to phorbolesters. The absence of RA in RAG and BC treated cells gives strong evidence that RAG and BC exert intrinsic biological effects. PMID- 9350479 TI - Carotenoids and intercellular communication via gap junctions. AB - Stimulation of gap junctional communication has been suggested to be one of the biochemical mechanisms underlying the cancer-preventive activity of carotenoids. The stimulatory activity is associated with structural properties of the carotenoids. It appears that the presence of a six-membered ring substituent at the end of the conjugated system of double bonds is required for the effects; five-membered ring carotenoids are less active. There is increasing evidence that oxidation products of carotenoids, especially retinoic acid analogs, significantly contribute to this biological property. PMID- 9350480 TI - Redifferentiation of oral dysplastic mucosa by the application of the antioxidants beta-carotene, alpha-tocopherol and vitamin C. AB - In a clinical trial the effect of chemoprevention with beta-carotene, vitamin E and C on dysplastic tissue was studied. The study included 24 patients with oral leukoplakia and 24 patients after radical resection of a primary oral cancer. There was a reduction of increased cell kinetic parameters like the S-phase portion or the average number of nuclear-organizer regions (NOR) per cell nucleus, a decrease of the micronuclei portion and a normalization of the cytokeratin gene-expression. The general response was 97.5%. Stopping the alcohol and tobacco abuse the effect of the antioxidative vitamins on redifferentiation of the oral mucosa was more intense than by persistance of the alcohol and tobacco abuse, but a long term prevention seems to be ineffective. PMID- 9350481 TI - Multiple carboxylase deficiency: inherited and acquired disorders of biotin metabolism. AB - Acquired biotin deficiency and the two known congenital disorders of biotin metabolism, biotinidase and holocarboxylase synthetase (HCS) deficiency, all lead to deficiency of the 4 biotin-dependent carboxylases, i.e. to multiple carboxylase deficiency (MCD). The underlying mechanism in HCS-deficiency, discovered in 1981, is decreased affinity of HCS for biotin impairing the formation of holocarboxylases at physiological biotin levels. In biotinidase deficiency, discovered in 1983, MCD results from progressive development of biotin-deficiency due to inability to liberate and recycle biotin which is lost in urine as biocytin. MCD leads to typical organic aciduria and severe life threatening illness. Main symptoms and signs are feeding difficulties, neurologic abnormalities (hypotonia, impaired consciousness, seizures, ataxia) and cutaneous changes (rash, alopecia). However, the clinical presentation and age of onset are extremely variable, and organic aciduria may initially be absent in biotinidase deficiency. Therefore, the definitive diagnosis requires enzyme studies. MCD can be detected in lymphocytes obtained before treatment and biotinidase deficiency is confirmed or excluded by a colorimetric enzyme assay in plasma. Newborn screening for biotinidase deficiency has resulted in the detection of patients with partial deficiency (10-30% of mean normal activity) in addition to patients with profound deficiency (0-10%). Severe illness has been observed mainly in patients with O-activity or a Km-mutation, detection of which requires detailed investigation. HCS-deficiency has to be confirmed by enzyme assay in cultured cells. Both congenital disorders respond clinically and biochemically to oral biotin therapy. Whereas 10 mg/day or less is sufficient to treat profound biotinidase deficiency, the optimal biotin dose for patients with HCS-deficiency must be assessed individually. The prognosis of both disorders is good if biotin therapy is introduced early and continued throughout life. However, delayed commencement of therapy in biotinidase deficiency can result in irreversible neurological damage, and in HCS-deficiency a few patients have responded only partially even to massive biotin doses of up to 100 mg/day. PMID- 9350482 TI - Enzymatic effects of folic acid and vitamin B12. PMID- 9350483 TI - Causes and consequences of hyperhomocyst(e)inemia. AB - Increased plasma concentrations of the sulfur-containing amino acid homocyst(e)ine are designated as hyperhomocyst(e)inemia. Various definitions have been used to derive cut-off levels for hyperhomocyst(e)inemia. The classification by Kang is now generally used distinguishing moderate, intermediate and severe hyperhomocyst(e)inemia. A variety of causes are discussed for the etiology of the disease which can be grouped into genetic and nongenetic factors. Severe hyperhomocyst(e)inemia is accompanied by homocystinuria and several other symptoms occurring early in life. Treatment is mandatory for normal development and prevention of premature atherosclerosis. Even less severe forms of hyperhomocyst(e)inemia imply a substantially elevated risk for vascular diseases. Etiology and severity of defect(s) leading to hyperhomocyst(e)inemia are the basis for treatment. In genetic defects, supplementation with the cofactor(s) of the affected enzyme is used to enhance enzyme activity. Alternative routes in the pathway may also be enhanced. Nongenetic hyperhomocyst(e)inemia often requires correction of suboptimal vitamin concentrations. Nutritive doses of the vitamins may be sufficient for treatment of less severe forms as well as for prevention of hyperhomocyst(e)inemia. PMID- 9350485 TI - The development of communication and language in deaf preschool children with cochlear implants. AB - The study is an ongoing longitudinal and qualitative psycho-social study of the communicative development in 19 preschool children with cochlear implants, using sign language. The children are video-recorded in natural interactional settings. Analysis of patterns of communication show that 16 of the children use sign language in communication with adults and peers. With regard to oral communication, 13 children were observed to utter single words or speech-like sounds upon an adult's request, but seldom used spoken words spontaneously. Six children used single spoken words in dialogues with adults if the content of the dialogue was about the here and now, and if the topic of reference was clear. None of the children in the study were able to take part in age-adequate play activities with peers when speech was used in communication. The results are discussed in reference to early mother-infant interaction, the development of communication and language, and the significance of early close relationships for children's social and emotional development. PMID- 9350484 TI - The impact of day care on ventilation tube insertion. AB - OBJECTIVE: To study the effect of day care and tube type, as well as other risk factors related to ventilation tube insertion and reinsertion. DESIGN: A case series of 456 consecutive cases with 162 controls from a well-baby examination group evaluated for age, sex, smoking history and day care attendance. SETTING: Private practice in a mid-sized, southeastern university town. PATIENTS AND CONTROLS: This was a referred sample of patients who were entered consecutively in a private-practice setting. All children were age five or less at entry into the study. INTERVENTIONS: Children underwent ventilation tube insertion with or without adenoidectomy, using standing indications. MAIN OUTCOME MEASURES: The primary outcome measures were whether or not a child had the need for a second set of tubes, and also determining the status of the child's ears for at least 1 year after tubes had extruded. RESULTS: Day care and younger age were both identified as risk factors associated with initial ventilation tube insertion. Only 11% (seven out of 63) of home care children, as compared with 31% (108 out of 346) day care children, had the insertion of ventilation tubes (P = 0.000). Day care children who had tubes inserted and adenoidectomy (with or without tonsillectomy) had a significantly lower rate of reintubation than children who had tube insertion alone (P = 0.00). Day care and young age are significant risk factors for any child, both with a first set of tubes and for ventilation tube reinsertion. Children in day care had a reintubation rate of 36% as compared to 11% for those in home care. Parents should be aware that day care can represent a two-fold hazard both in the observed connection between day care and tube insertion and the demonstrated increased probability of reinsertion. Any studies looking at ventilation tube outcomes need to make certain to monitor for day care attendance. PMID- 9350486 TI - Intranasal endoscopic silicone intubation for congenital obstruction of the nasolacrimal duct in children. AB - Aim of the present study was to determine the efficacy of intranasal endoscopic silicone intubation to treat congenital nasolacrimal duct obstruction in children who failed conventional lacrimal system probing. Silicone intubation with intranasal endoscopic visualization was performed in 18 eyes of 16 patients with an age range of 18-48 months (mean, 25 months) to treat congenital nasolacrimal duct obstruction. Out of 16 patients, ten had already undergone previous probings with failure, the other six were primary intubation cases without any previous probings. The tubes were left in place an average of 6 months. Follow-up ranged from 4 months to 2 years (mean, 12 months) following removal of the silicone tubing. Intranasal endoscopic silicone intubation is effective in the treatment of congenital nasolacrimal duct obstruction as a primary procedure in children over 18 months of age and, it is also recommended as the next step in the management of epiphora which fails to resolve after probing. PMID- 9350487 TI - Influence of hyperoxia on in vitro growth of rabbit middle ear epithelium and auditory meatal epithelium. AB - The oxygen partial pressure of middle ear gas increases more than 3-fold upon insertion of ventilation tubes, while the carbon dioxide partial pressure decreases. Whereas the middle ear gas is normally equilibrated to venous gases and has an oxygen partial pressure of 43 mmHg, 138 mmHg is measured in ventilated ears. The present study was undertaken to compare the effects of these oxygen tensions on in vitro growth and glycoprotein secretion of rabbit middle ear epithelium and for comparison auditory meatal epithelium. Cultures were incubated in atmospheres of 7, 21 or 75% O2 in 5% CO2 and the remnant N2. The cell layer protein mass, [3H]thymidine-incorporation, DNA content and [3H]glucosamine incorporation was measured in identical subcultures every third day during a 15 day period. In middle ear epithelium the DNA content, DNA synthesis and cell layer protein mass were significantly higher at 7% oxygen compared to 21% and 75%. In conclusion hyperoxia leads to decreased growth of middle ear epithelium in vitro. If applicable to in vivo conditions, this might contribute to the mechanism of action of ventilation tubes. Moreover the proliferation rate of auditory meatal epithelium exceeds that of middle ear epithelium both at 7 and 21% oxygen, an interesting point with regards to cholesteatoma pathogenesis. PMID- 9350488 TI - Videonasopharyngoscopy as an instrument for visual biofeedback during speech in cleft palate patients. AB - Videonasopharyngoscopy was used as an instrument for visual biofeedback during speech in cleft palate patients. Seventeen cleft palate patients were randomly selected for the study. All patients showed velopharyngeal insufficiency (VPI), compensatory articulation (CA) and negative movement of lateral pharyngeal walls (NMLPW) during speech. Nine patients received speech therapy for correcting CA. Eight patients received speech therapy and underwent videonasopharyngoscopy as an instrument for visual biofeedback of the velopharyngeal sphincter. After 12 weeks, NMLPW was modified in the patients receiving speech therapy and visual biofeedback. In contrast, NMLPW was still present in eight out of nine patients receiving only speech therapy. These patients received visual biofeedback and NMLPW was corrected in all cases. After six months, all 17 patients had corrected CA during isolated speech. All patients received a tailor-made pharyngeal flap. VPI was completely corrected in 15 cases. In the two cases in which VPI was still present postoperatively, the size of the defect at the velopharyngeal sphincter had been significantly reduced. In these two patients, visual biofeedback was used postoperatively for increasing lateral pharyngeal walls (LPW) motion towards the borders of the flap. After 18 months since the onset of speech therapy all the patients had normal nasal resonance and normal articulation during connected speech. PMID- 9350489 TI - Three-dimensional imaging of the pediatric airway. AB - Accurate imaging of the pediatric tracheobronchial tree is indicated for the evaluation of congenital or acquired abnormalities. Conventional axial computed tomography (CT) is now considered the best imaging modality for evaluation of the trachea and major bronchi, and has almost completely replaced the former gold standard of tracheobronchography. Preliminary results indicate that CT scan performance is further enhanced through the application of spiral technology and two-dimensional (2D) and 3D representation of the tracheobronchial tree. Spiral CT scans with 3D surface rendering offers an opportunity to replace traditional tracheobronchography with a safer, less invasive modality. PMID- 9350490 TI - Pediatric tonsillectomy: post-operative morbidity comparing microsurgical bipolar dissection versus cold sharp dissection. AB - A prospective single-blinded study was conducted to compare pediatric tonsillectomy using the traditional cold sharp dissection with ligation for securing hemostasis (CSDL) versus microsurgical bipolar dissection technique (MBCT). Sixty children aged between 2 and 14 years were sequentially assigned to each group. Blood loss and postoperative pain in the first hour were markedly decreased in the MBCT group (P < 0.0001 and P < 0.05, respectively). Average surgical time was slightly decreased in diathermy tonsillectomy (15'30") compared with the cold technique (16'30"). Return to normal diet was significantly earlier in the CSDL group (P < 0.05). Late postoperative pain measured on the tenth day showed a moderate increase in children who underwent MBCT. There was no difference between the two techniques in the incidence of postoperative hemorrhage. No major complications occurred. We believe MBCT is a fast and safe technique which may be useful in children with known bleeding disorders. However, it increases late postoperative pain and, therefore, delay in patients' return to normal activities. Consequently, it does not represent a significant advantage over the traditional CSDL procedure. PMID- 9350491 TI - Acoustic rhinometry in the evaluation of congenital choanal malformations. AB - Previous studies have shown that acoustic rhinometry (AR) is well suited to describe the nasal airway dimensions in healthy infants. The technique is quick to perform, non-invasive, without potential hazards and requires minimal cooperation. Due to the small dimensions of the infant nasal airways, the optimized miniprobe (Rhinometrics, Lynge, Denmark) provides abilities superior to those of adult probes. The equipment is portable and examinations can be performed in the maternity ward, out-patient department or at home. Measurements on models simulating the pre- and post-operative nasal geometry were performed to determine the accuracy and resolution of AR and to improve the interpretation of the curves obtained in vivo. Acoustic measurements in five infants with congenital respiratory distress caused by bilateral choanal atresia or stenosis have been compared with CT-scans and the results of the model simulations to evaluate the diagnostic value of the method. We conclude that AR represents a new and valuable tool in the diagnosis of congenital choanal malformations. Despite current technical limitations, it may also be of value in the post-operative evaluation, particularly if the posterior part of the septum is not resected. PMID- 9350492 TI - Hearing loss associated with neonatal ECMO: a clinical investigation. AB - To answer the question as to the prevalence of sensori-neural hearing loss (SNHL) among neonates receiving ECMO, a retrospective chart review was conducted on 198 infants having surgery between November 1987 and January 1995. One hundred fifty seven (79.7%) survived. One hundred thirty infants met our criteria of having a pre-discharge auditory brainstem evoked response (ABR) test and at least one follow-up behavioral audiologic examination. Strict criteria were set for normal hearing on both the ABR and follow-up examinations. Only follow-up results are reported. At the time of the most recent follow-up examination, two children could not be adequately studied, 106 exhibited normal hearing, 21 (16%) exhibited either unilateral or bilateral conductive hearing loss and three (2.3%) exhibited unilateral or bilateral SNHL. Only one child is using amplification. With the largest sample size to date, we found a lower prevalence of SNHL after ECMO than has been previously noted in the literature. Although the prevalence of hearing loss is low, the post-ECMO group of infants must be considered at risk for hearing loss. The prevalence of hearing loss cannot be based solely on a pre discharge ABR, i.e. ongoing follow-up testing is necessary. PMID- 9350493 TI - Non-endoscopic techniques for the evaluation of the pediatric airway. AB - In children with stridor, a detailed evaluation of the airway is often required to assess precisely its anatomical and functional status. Various methods of assessment have been developed and airway management may include, as well as rigid and flexible endoscopy, the use of imaging techniques such as plain X-rays, a barium oesophagogram, ultrasound, a CT scan, a magnetic resonance image (MRI) and an angiogram, as well as respiratory function tests including acoustic rhinometry and flow volume loops or even pH monitoring. This article aims to highlight the valuable information these alternative techniques can provide. PMID- 9350494 TI - The use of the salivagram in the evaluation of severe and chronic aspiration. AB - Chronic salivary aspiration may be responsible for a significant percentage of pneumonia in the neurologically impaired child. The radionuclide salivagram (RS), a simple investigative study, can document salivary aspiration as the source of pulmonary contamination. The purpose of this study was to determine if the results of the RS would accurately identify children with severe and chronic salivary aspiration who would benefit from laryngotracheal separation (LTS). We reviewed 30 records of children with chronic aspiration pneumonitis who underwent LTS between 1988 and 1996. We recorded the number of inpatient days required for respiratory infections before and after LTS. This number was compared with the number of inpatient days for respiratory infection from children (n = 27) who underwent the RS during a ten-month period but who were never referred for LTS. Fifteen children who underwent LTS had a preoperative RS. The RS documented salivary aspiration in 11 of these children. Aspiration was effectively controlled by LTS for this group. There were three studies that failed to show either aspiration or progression of the Technetium 99m sulfur colloid (Tc 99m SC) into the esophagus This finding was felt to represent significant swallowing dysfunction and, therefore, was also considered a positive finding. There was a significant difference in the number of inpatient days for children who had a negative RS and were never referred for LTS when compared with the number of inpatient days for those children who had a positive RS and were referred for LTS. We feel that the RS is an effective tool to document salivary aspiration as the source of recurrent pneumonia. A modification of the technique and interpretation of RS is suggested. PMID- 9350495 TI - Botryomycosis: improved therapy for a difficult infection. AB - Botryomycosis is a chronic bacterial granulomatous disease often involving the skin and subcutaneous tissue. Head and neck involvement is rare. Botryomycosis presents with clinical and histological features similar to actinomycosis or mycetoma, but the causative organism is usually Staphylococcus aureus. Microscopically the organisms appear to be encapsulated in granules, which are thought to protect them from the effects of standard courses of antibiotics. Botryomycosis usually requires surgical intervention for cure. Major debilitating surgery has been required for most patients, because the infection has been unresponsive to seemingly appropriate medical therapy. We present an 8-month-old male with periorbital botryomycosis. Surgical specimens for diagnosis were obtained, but complete resection would have created debilitating functional and cosmetic defects. The lesion failed to respond to nafcillin alone or combination therapy with hyperbaric oxygen, but showed slow, steady improvement with long term clindamycin. The patient has been disease free for more than 4 years, with minimal scarring and no functional impairment. Prolonged medical therapy for botryomycosis may be a viable alternative to the traditionally recommended surgical resection, thereby reducing cosmetic and functional morbidity. PMID- 9350496 TI - Primitive pharyngeal dyskinesia associated with upper airway collapse in an infant. AB - We report a case of total collapse of the upper pharyngeal airway in a slightly premature baby, resulting in a noisy breathing disorder. Primary immaturity of the central nervous system contributing to pharyngeal muscle hypotonia has been implicated in association with the increase in nasal pressure. The infant experienced complete resolution of symptoms a few weeks after the placement of a nasopharyngeal tube. This case report demonstrates the difficulty in diagnosis and management. The developmental spectrum and exploration are reviewed. PMID- 9350498 TI - Cardioexcitatory neurons in the snail Achatina fulica. AB - The excitatory motor unit of Achatina fulica is composed of five identified cardioregulatory motor neurons: three tonically active neurons (TAN-1, -2 and 3), a periodically oscillating neuron (PON) and a VG1 neuron. High frequency discharges in TAN neurons evoked slow depolarization waves. The PON elicited biphasic excitatory post synaptic potentials (EPSP). One spike in PON was sufficient to increase heart rate for several minutes. VG1 elicited discrete fast EPSP in the myocardium. Bursts of spikes from VG1 resulted in a summation of EPSP and transiently increased heart. VG1 inhibited spontaneous electrical activity in PON. Our results suggest that the use of semi-intact preparations allow elucidation of new functional cardioregulatory properties of intact neural networks. PMID- 9350497 TI - Cardiac inhibitory neurons in gastropod snail Achatina fulica. AB - Two inhibitory heart motor neurons (HI-1, HI-2) were identified in the visceral ganglion of the snail Achatina fulica. Both motor neurons were connected monosynaptically with the myocardium. Irregular action potentials composed a typical pattern of neuronal spontaneous electrical activity. The neurons shared common excitatory synaptic inputs, and were connected through electrical synapses. Neuronal endings of these cells responded to tactile stimulation and to cardiac stretching when the chemical synapses were blocked. HI-1 and HI-2 neurons both elicited inhibitory post synaptic potentials in the myocardium with a constant delay and thereby modulated cardiac spikes frequency and amplitude. Staining with the dye Lucifer yellow revealed that these two neurons were unipolar cells which had dendrites only in the visceral ganglion. PMID- 9350499 TI - Localisation of glutamate receptors in the substantia nigra pars compacta of the monkey. AB - The distribution of ionotropic glutamate receptor subunits GluR1, GluR2/3 and NMDAR1, and meta-botropic receptor mGluR1 alpha was studied in the monkey substantia nigra. High levels of immunoreactivity to GluR1, GluR2/3 and NMDAR1, and moderate levels of immunoreactivity to mGluR1 alpha were observed in the substantia nigra pars compacta. GluR1 and GluR2/3 were mostly in cell bodies and larger stem dendrites, whilst NMDAR1 and mGluR1 alpha were present on medium sized and small dendrites, respectively. The substantia nigra receives glutamatergic afferents from the subthalamic nucleus and the frontal cortex. Overactivity of the subthalamic nucleus, coupled with high levels of glutamate receptors on the neurons in the pars compacta, could predispose these neurons to excitotoxic injury, and could contribute to the development of Parkinson's disease. PMID- 9350500 TI - Projections from the nucleus isthmi to the visual and auditory centres in the frog, Rana esculenta. AB - The lectin Phaseolus vulgaris leucoagglutinin was injected into the nucleus isthmi (NI) in order to study its anterograde and retrograde projections in the frog. The following areas of termination could be discerned in the brainstem: (1) Each of the five subnuclei of the torus semicircularis (TOS) received fibres from the NI. The projection was the most extensive on the three main subnuclei which disclosed also retrogradely labelled neurones on the side of injections. The subependymal subnuclei contained the least number of labelled fibres. (2) Both hemispheres of the optic tectum (TO) were supplied by fibres from the NI. Labelled fibres were more numerous on the side of injections, and preterminal and terminal fibres covered columnar-like areas in layers 8 and 9. Several retrogradely labelled neurones were found in layer 6. Relatively few labelled fibres were seen on the contralateral side. They formed patch-like areas of termination in layer 9. (3) The anterodorsal (AD) and anteroventral (AV) nuclei were reciprocally inter-connected with the NI. The fibre connections were less extensive on the contralateral side. In the rhombencephalon (4) the cochlear nucleus (CN) and (5) the superior olive (SO) were also reciprocally connected with the NI on both sides, but with much weaker projection on the side contralateral to injections. (6) Only a weak anterograde labelling was observed in the contralateral NI and in the ipsilateral reticular formation. PMID- 9350501 TI - Do microglial cells have a neuroprotective function? AB - To explore the possible neuroprotective role of microglia after a peripheral nerve lesion, the present study used athymic mice shown to have a lower number of microglia to highlight some functions of microglia not normally obvious in animals with a normal number of such cells. Observation of semithin sections showed that unoperated 5-day-old athymic mice had a significant lower number of microglial cells around sciatic motoneurons, compared to their BALB/c littermates. After right sciatic nerve cut at mid-thigh level, motoneuron loss occurred faster at 5 and 10 days after operation in neonatal athymic mice than BALB/c mice. The motoneuron loss by 15 days after nerve cut was, however, the same in both strains of mice. Microglial reaction after sciatic neurectomy, as revealed by Mac-1 immunohistochemistry, was obviously less in intensity and number in athymic mice than in BALB/c mice. The results indicated a neuroprotective function of microglia, which, when not present in adequate numbers, could result in a faster motoneuron death. The study also showed that while microglia were reduced in number in athymic mice, there was no significant difference in the number of astrocytes and oligodendrocytes in the spinal cord of the athymic mice, compared to that in BALB/c mice. This indicates that the development of astrocytes and oligodendrocytes in the spinal cord may not be affected by a reduced number of microglia. PMID- 9350502 TI - Group I metabotropic glutamate receptor agonist causes neurodegeneration in rat hippocampus. AB - Activation of group I metabotropic glutamate receptors results in an increase in cytosolic Ca2+. High levels of free Ca2+ in the cytoplasm could be toxic to cells. We now report that intracerebroventricular injections of a group I metabotropic glutamate receptor agonist (S)-3,5-dihydroxyphenylglycine (DHPG) leads, within hours, to changes in the distribution of ionotropic glutamate receptors in the hippocampus and a delayed (4-7 days) loss of hippocampal pyramidal neurons. Injection of glutamate transporter substrates resulted in no loss of neurons and the administration of an NMDA agonist produced lesions different from those caused by (S)-DHPG. These results suggest that the activation of metabotropic glutamate receptors of group I in vivo may indeed result in neurotoxic events. PMID- 9350503 TI - Species differences in the localisation of gamma-glutamyl transpeptidase immunopositive cells at the blood-brain interface. AB - The expression of gamma-glutamyl transpeptidase (GGT) has been associated with the emergence of a functional blood-brain barrier. We have undertaken a precise localisation of this enzyme in the cerebral cortical vessels of different species, by immunocytochemistry using a polyclonal antibody and light and electron microscopy. GGT immunoreaction product was present on the luminal surface of endothelial cells in 1-day-old to 3-month-old rats, whereas in the mouse, monkey and human cortex, this protein was present in astrocytic endfeet around the vessels. No labelling was observed in the other cellular components of the vessel walls, such as pericytes, fibroblasts, smooth muscle cells and perivascular cells. The localisation of GGT in astrocytes in mice, monkeys and humans suggests that these cells could play a role in the detoxication of lipophilic xenobiotic substances that cross the endothelial barrier. In these species, astrocytes can be viewed as a second line of defense against xenobiotics, beyond the capillary endothelium. PMID- 9350504 TI - Increase in extracellular calcium in the optic tectum of fish after optic nerve transection. AB - In this study the extracellular distribution of cytochemically generated calcium reaction product in the denervated optic tectum of a cichild fish (Oreochromis mossambicus) was investigated. The left optic nerve had been transected and the fish (5 per experimental condition) maintained for 2, 10 and 21 days. The amount of the calcium-containing precipitates was estimated using energy-filtering transmission electron microscopy (EFTEM) and image analysis. A special degeneration type of the optic terminals (neurofibrillar hypertrophy) was found which seems to be rare in other teleosts and was therefore chosen for quantification of the calcium deposits. These terminals are surrounded by astroglial processes and the calcium reaction product in the extracellular spaces between these glial processes and the terminals was measured and compared to normal optic terminals in nonoperated controls. A distinct and significant increase in the amount of calcium deposits was found 2 and 10 days after surgery which decreased to control levels after 21 days. This rise of deposits around the degenerating terminals was very local as arbitrarily selected extracellular spaces near these terminals showed values which were at the level of the nonoperated controls. Therefore, a transient and local increase in extracellular calcium precipitates was found after optic nerve transection which affected only the degenerating synapses. PMID- 9350505 TI - Structure of glycogen particles in organ of Corti's outer hair cells in three rodent species. AB - Organ of Corti's outer hair cells are one of the few cell types in mammals to contain large cytoplasmic glycogen stores, and the only one in the adult auditory receptor. Previous reports on the structure and distribution of glycogen in the adult organ of Corti were mainly based on light microscopy histo- and cytochemical methods, and the scare EM studies on the topic relied on techniques which were not sensitive or specific enough. Furthermore, it has been reported that glycogen particles are not present in outer hair cells of all species. A first goal of the present study was to describe the ultrastructure of glycogen stores in organ of Corti's outer hair cells in Guinea pig, rat, and mouse, using the periodic acid-thiocarbohydrazide-silver proteinate method. In addition, differences in the subcellular and cochleotopic distribution of this substance were analyzed. In the adult organ of Corti only the outer hair cells contain glycogen stores. Present throughout their cytoplasm, these deposits appear either as single beta particles, or as aggregates of these, forming alpha particles. Though most alpha particles are round, some appear long and conspicuously straight in longitudinal sections of those cells near the apex of the cochlea, and they seem to be apposed to some filamentous structure. On the other hand, when the cells are sectioned transversely the larger aggregates of glycogen particles follow a curved course. Since outer hair cells of the apical region of the cochlea contain a bundle of contractile microfilaments, our results suggest that glycogen is associated with the contractile apparatus of these cells. This hypothesis is in good accordance with previous experimental data which suggest that glycogen is used as energy source for the contractile movements of outer hair cells. PMID- 9350506 TI - Distribution of amyloid beta-protein immunoreactivity in the hippocampus of rats injected with kainate. AB - The distribution of amyloid beta-protein immunoreactivity was investigated in the hippocampus of rats injected intravenously with kainate. Very light labelling was observed in cell bodies and dendrites of pyramidal neurons and dentate granule cells in the normal hippocampus. At 6,7,9,10,12 and 20 days postinjection, moderately densely labelled astrocytes were present in the stratum oriens and lacunosum moleculare, extending thick processes towards the stratum radiatum which was degenerating. Immunoreactivity was present as floccules in the mitochondria-rich but glial-filament-poor portions of reactive astrocytes, and in fibrillar form in the neuropil. This suggests that amyloid beta-protein might be present in fibrillar (presumably polymerised) form in degenerating profiles and the neuropil, but in floccular (presumably non-polymerised) form in reactive astrocytes. PMID- 9350507 TI - Neurokinin B receptor (NK3)-positive neurons expressing c-fos after chemical noxious stimulation on the peritoneum: a double staining study in the nucleus tractus solitarius of the rat. AB - Neurokinin B receptor (NKR; NK3)-like immunoreactivity (LI) was densely distributed in the nucleus tractus solitarius (NST) of the rat. Approximately 32% of the neurons with NKR-LI in the NST expressed c-fos protein after chemical irritation on the peritoneum with acetic acid. The present results suggest that neurons containing NKR in the NST may be involved in the integration of noxious afferent information from the peritoneum in the rat. PMID- 9350508 TI - Differential changes in the expression of AMPA receptors genes in rat brain after chronic exposure to ethanol: an in situ hybridization study. AB - Chronic treatment with ethanol of adult rat was used as an experimental paradigm to investigate changes in the expression of glutamate receptor genes under the condition of a slowly progressing neurodegeneration that is associated with a plastic neuronal growth response. Effects of chronic oral ethanol exposure (20% v/v, 28 weeks) on the expression of the AMPA receptor mRNA subtypes GluR-A, GluR B and GluR-C (flip variants) were analysed in the rat hippocampus by in situ hybridization and subsequent quantitative autoradiography. In both controls and ethanol treated animals strong hybridization signals could be detected over pyramidal neurones and dentate gyrus cells. GluR-C mRNA was elevated by 15% to 30%, while expression of the AMPA receptor transcripts GluR-A and GluR-B were not significantly altered after ethanol treatment. The present findings demonstrate that chronic ethanol exposure affects gene expression of AMPA receptor subtypes differentially. Differential influences of distinct AMPA-receptor subtypes on neuronal survival and plastic neuronal response under conditions of chronic neuronal degeneration might, therefore, be suggested. PMID- 9350509 TI - Antibody to keyhole limpet hemocyanin labels retinal horizontal cells in some amphibians, but not in others. AB - Antibody to keyhole limpet hemocyanin (KLH) reacts with putative horizontal cells in anuran amphibians of the superfamily Bufonoidea. The reactive epitope appears to be located on the cell membrane. No KLH-like immunoreactivity was observed in the outer plexiform layer (OPL) of anurans not members of this superfamily, nor in the OPL of urodeles or other vertebrates. Thus KLH-like immunoreactivity in the OPL provides a tool for assessing phylogenetic relationships within anurans. PMID- 9350510 TI - On the role of Muller glia cells in histogenesis: only retinal spheroids, but not tectal, telencephalic and cerebellar spheroids develop histotypical patterns. AB - The establishment of cell and fibre layers and the specification of different cell types are crucial processes during development of the central nervous system. Here we investigated the developmental architecture of radial glia cells in these processes using so-called spheroids that arise from dissociated chicken embryonic neural cells in rotation culture. We were able to produce retinal, tectal, and telencephalic spheroids from E6 embryos and cerebellar spheroids from E10 embryos. Cell and fibre differentiation can be observed in all types of spheroids, however, it is most abundant in retinal spheroids. Moreover, only in retinal spheroids a histotypic organization can be detected. Using immunohistochemistry and electron microscopy, we assign this -at least partially- to the capacity of Muller cells to form radial scaffolds, since we observe a congruency between these radial scaffolds and the presence of rosettes formed by photoreceptor precursors and Muller cells. Tectal, telencephalic and cerebellar spheroids do not show organized radial glia scaffolds, instead, the radial glia cells are randomly arranged and the spheroids do not show histotypical organization. The application of the specific gliotoxin 6-aminonicotinamide to growing retinal spheroids leads to a significant decrease in the number and size of the rosettes. Concomitantly, the degree of histotypical organization is also drastically reduced. This organizing capacity of Muller cells in vitro now strongly suggests the presence of a comparable function also in vivo. Moreover, since non-retinal radial glia cells are not able to re-organize an histotypic organization in vitro, Muller cells seem to be qualitatively different from other radial glia cells. In future studies we want to untangle these differences. PMID- 9350511 TI - Histological visualization of long fiber tracts in the white matter of adult human brains. AB - The position and extent of fiber pathways and their intersubject variability in the white matter of the forebrain have not been analyzed with histological techniques in the adult human brain. This is due mainly to the lack of a staining procedure with which the different fiber tracts can be visualized with sufficient contrast. The here described modification of the Heidenhain-Woelcke myelin stain yields increased contrast between heavily and less heavily myelinated pathways, facilitating identification and tracing of single fiber tracts. The modification makes the microscopical resolution possible which is necessary for preparing comprehensive maps of fiber tracts. Such maps are valuable tools for clinical diagnosis, since they provide a basis for localizing anatomically the structures involved in subcortical lesions. Correlations between the size of a lesion and its location relative to identified fiber tracts on the one hand and postlesional functional alterations and recovery on the other can only be made on the basis of topographic and morphometric maps of these structures in the normal adult human brain. PMID- 9350512 TI - Anterograde projections of the cortical tongue area of the tree shrew (Tupaia belangeri). AB - In altogether seven tree shrews, Tupaia belangeri, the anterograde projections of the motorcortical tongue area were investigated as part of a larger comparative study. Identification of the tongue area was carried out by electrical brain stimulation. Three different tracers were used: biotin-conjugated dextranamine (BDA), Phaseolus vulgaris leucoagglutinin (PHA-L) and 3H-leucine. Intracortical projections were found to the motor cortex around the injection site, the premotor cortex, supplementary motor area, the homologues of the primate frontoparietal operculum and insula, the anterior cingulate and agrannular retrosplenial cortex, the somatosensory and bordering inferior parietal cortex as well as to the perirhinal cortex. Except the very weak projections into the retrosplenial, posterior parietal and perirhinal region which were ipsilateral, all other projections were bilateral. Subcortically, there was a projection into the ventral putamen, rostrodorsal claustrum and, very sparsely, into the caudate nucleus. In the thalamus, terminal labeling was found in the nuclei reticularis, anteroventralis, anteromedialis, ventralis lateralis, ventralis posterior medialis, ventralis posterior inferior, medialis dorsalis, in the intralaminar nuclei paracentralis, centralis lateralis, centrum medianum and parafascicularis, in the midline thalamus and in the nuclei posterior and pulvinaris. Further diencephalic projections, however all of them wak, could be traced into the zona incerta, dorsolateral subthalamus, dorsomedial, lateral and supraoptic hypothalamus. In the midbrain, labeling was found in the deep layers of the lateral superior colliculus, in the bordering reticular formation and, very sparsely, in the periaqueductal grey. In the lower brain-stem, fibres ended in the griseum pontis, dorsolateral reticular formation, principal and spinal trigeminal nucleus and, sparsely, in the lateral parabrachial region, solitary tract nucleus, inferior olive and magnocellular reticular formation. No terminals were found in the hypoglossal nucleus. The projection system revealed with PHA-L was less extensive than that demonstrated with BDA and 3H-leucine, both of which were similar. PMID- 9350513 TI - Combined amplification and sequencing in a single reaction using two DNA polymerases with differential incorporation rates for dideoxynucleotides. AB - We describe an approach, which combines the process of DNA amplification and sequence determination by using a pair of primers and two DNA polymerases with different incorporation rates for dideoxynucleotides. The process of target sequence amplification is carried out by the DNA polymerase with a low dideoxynucleotide incorporation rate while its polymerase counterpart with a high incorporation rate generates a sequence ladder. The needs for separate amplification via polymerase chain reaction (PCR) or cloning into plasmids including the respective purification steps therefore can be avoided. In addition, the use of dye terminator chemistry enables the simultaneous generation of forward and reverse sequence ladders, which can be separated based on the streptavidin-biotin system when one amplification primer is biotinylated. PMID- 9350514 TI - Ussing chamber for high-frequency transmural impedance analysis of epithelial tissues. AB - An Ussing chamber was designed for impedance analysis of epithelial tissue and optimized for the use of high-frequency alternating current stimuli. Shielded voltage electrodes, located axially within the electric field of the Ussing chamber, minimized the reactive properties of the set-up. By vectorial subtraction, the small reactive contribution of the optimized Ussing chamber was completely compensated for. This method allowed for transmural impedance measurements in a frequency range of 1-65 kHz. For the first time, Nyquist plots of cultured intestinal cell monolayers (HT-29/B6) are presented. The epithelial monolayers in different stages of confluence showed the impedance locus as a semicircle, with the high frequency end close to the origin. These epithelial monolayers could be modeled by a simple RC-parallel circuit without a series resistance. PMID- 9350515 TI - Manipulation and trapping of sub-micron bioparticles using dielectrophoresis. AB - A non-uniform alternating electric field induces motion in polarisable particles called dielectrophoresis. The effect is governed by the relative magnitudes of the dielectric properties of the medium and the particles. The technology has been used to manipulate particles for biotechnological applications, including purification, fractionation and concentration of cells and microorganisms. However, the lower size limit for the dielectrophoretic manipulation of particles was believed to be about 1 micron, but recent work has proved otherwise. The dielectrophoretic movement and properties of latex beads and a simple rod-shaped virus, tobacco mosaic virus (TMV), have been measured using microfabricated electrode structures. Measurements have been made over a range of suspending medium conductivities, applied frequencies and electric field strengths. It is shown that under appropriate conditions both latex beads and tobacco mosaic virus particles can be selectively attracted to regions of high electric field strength located at the tips of microfabricated electrode structures. The ability to selectively trap and separate bio-particles has many potential applications in the area of biotechnology. PMID- 9350516 TI - A photo microcalorimetric system for studies of plant tissue. AB - A gas perfusion microcalorimeter, primarily designed for studies of plant tissue, has been equipped with light guides in order to allow studies under the influence of light (UV-vis), and with variations in gas phase composition. The calorimetric system consists of three twin microcalorimeters of the heat conduction type positioned in a thermostated water bath. The biological sample is placed in a 20 ml photo calorimetric vessel. Light is introduced by means of a quartz rod connected to one of the arms of a bifurcated light cable. The other arm of the cable is connected to a second microcalorimeter, serving as the photo calorimetric reference. The gas flow leaving the main calorimeter will pass the reaction vessel of the third microcalorimeter, where CO2 is trapped by NaOH solution. The measurement with this calorimeter will lead to values for the rate of uptake/release of CO2 by the biological sample. Test experiments have been conducted with pieces of spinach leaf, and values have been derived for the uptake of light energy at different concentrations of CO2. From the calorespirometric results a value was derived for the Gibbs energy of formation of photosynthate from CO2 and H2O. PMID- 9350518 TI - Sialic acid measurement by a modified Aminoff method: a time-saving reduction in 2-thiobarbituric acid concentration. AB - Our results show that if the 2-thiobarbituric acid concentration is decreased, its co-precipitation with the chromophore is diminished. Subsequent running of this reaction mixture by high-performance liquid chromatography still allows measurement of Neu-5-Ac in the picomole order, with a substantial time and reactive saving, as compared with the original assay. PMID- 9350517 TI - Quantitative determination and reversible modification of thiols using imidazolidine biradical disulfide label. AB - Earlier we reported an ESR method of quantitative determination of sulfhydryl groups. The method is based on the application of the imidazoline biradical disulfide label, R1S-SR1, which participates in the reaction of thiol-disulfide exchange followed by dramatic changes in ESR spectra. One of the disadvantages of the application of R1S-SR1 at physiological conditions is the requirement of excess of the biradical compared with thiol content which results in the consumption of the thiols and irreversible damage of the system under study. In the present paper we propose imidazolidine biradical disulfide reagent, R2S-SR2, for ESR determination of thiols and provide an experimental basis for its application. This label has the advantages of the previously used biradical disulfide, R1S-SR1, such as high sensitivity down to 1 microM of thiols even in opaque samples and could possibly be used for reversible modification of proteins and enzymes. The particular properties of the R2S-SR2 are pH-sensitivity of its ESR spectrum, higher stability of the imidazolidine radical fragment towards biological reductants and low concentration of the label sufficient for thiol determination at physiological conditions. The latter makes it possible to use ESR spectroscopy for non-invasive thiol measurements in biological systems, in vivo applications included. PMID- 9350519 TI - Time differential perturbed angular correlation of 111In-EDTA linked to the single free cysteine of bovine serum albumin. AB - By means of a facile chemical modification of the bovine serum albumin molecule, it was possible to measure its hydrodynamic radius with high accuracy (approximately 3%) using the TDPAC technique. The new approach presented here allows a wide use of the TDPAC technique to perform high precision studies of backbone dynamics of almost any protein. PMID- 9350554 TI - Plaque removal with variable instrumentation. AB - The purpose of this study was to evaluate dental plaque removal in a normal healthy mouth, during routine oral hygiene appointments using different techniques and without the use of any disclosing agents. 12 dental hygienists, randomly selected from a continuing education course, were asked to perform oral hygiene on the same patient to remove all the supra-gingival plaque without any time restriction and without the use of a disclosing agents. The plaque index score (O'Leary) was assessed before and after each session with the use of fluorescine and UV light source by an independent examiner. 3 groups of instruments were utilized: group A: ultrasonic scalers + prophy cups; group B: ultrasonic scalers + prophy cups + dental floss; group C: Gracey curettes + prophy cups. While no group was able to remove all the plaque, groups B and C performed significantly better. PMID- 9350555 TI - The optimization of the BANA test as a screening instrument for gingivitis among subjects seeking dental treatment. AB - Porphyromonas gingivalis, Treponema denticola and Bacteroides forsythus have been implicated in periodontal disease and each possesses an enzyme capable of hydrolyzing the synthetic trypsin substrate, BANA. We have used a chairside test for BANA hydrolysis to diagnose an anaerobic periodontal infection in patients with advanced forms of clinical disease using a 15-min/55 degrees C incubation protocol. However, the BANA test performance is dependent upon the length and temperature of incubation. In the present study, we have evaluated a 5-min/35 degrees C, a 5-min/55 degrees C and a 15-min/55 degrees C incubation protocol to determine whether the performance of the BANA test could be optimized using plaque samples obtained from subjects seeking dental treatment. Logistic regression models were tested with age, smoking status, and gingivitis scores as covariates. The best fitting model obtained with the 5-min/35 degrees C protocol had a sensitivity of 71%, a specificity of 68%, a false-positive proportion of 9%, a false-negative proportion of 65%, and an overall accuracy of 80%. When maximum likelihood estimates were obtained in this model, plaques from individuals who reported that they currently smoked were 9.57x, and those who quit smoking were 4.73x more likely to have a positive BANA score than someone who never smoked. Plaques were 4.55x more likely to be BANA-positive if they were removed from sites with gingivitis. These findings indicate that the performance of the BANA test is best using the 5-min/35 degrees C incubation protocol. PMID- 9350556 TI - Alveolar bone anatomic profiles as measured from dry skulls. Clinical ramifications. AB - The purpose of this study was to evaluate the relationship of alveolar bone morphology to tooth shape and form. 111 dry skulls were evaluated at Baylor College of Dentistry (Dallas, Texas). The skulls were arbitrarily divided into flat, scalloped and pronounced scalloped anatomic profiles according to alveolar bone anatomy. The number of buccal dehiscences and fenestrations was determined for each skull according to their anatomic morphotype. 10 skulls from each group were selected for bone height measurements. The measurements were made with a periodontal probe and ruler from the height of the interproximal bone to the buccal alveolar crest. Kodachrome slides were used to measure mesial-distal tooth width and length from ten skulls from each anatomic category. The average number of fenestrations for each group was 3.5. The mean number of dehiscences for flat and scalloped skulls was 0.5. The average number of dehiscences for pronounced scalloped was 1.2. There were no significant differences when the groups were compared. The mean distance from the height of the interdental bone to the alveolar crest was statistically significant when the groups were compared (flat 2.1 mm, scalloped 2.8 mm, pronounced 4.1 mm) (Tukey, p = 0.05). There were no significant differences when tooth shapes were compared with bone anatomy. Pronounced scalloped anatomic profiles were slightly narrower when compared with the other groups. The observations reported have treatment ramifications when patients with scalloped or pronounced scalloped morphotypes are being considered for dental implant placement. PMID- 9350557 TI - A short-term clinical study design to investigate the chemical plaque inhibitory properties of mouthrinses when used as adjuncts to toothpastes: applied to chlorhexidine. AB - The removal of plaque by toothbrushing with toothpaste is the most common form of plaque control in the developed world. However, the use of chemical adjuncts such as mouthrinses is increasing. In practice mouthrinses and toothpaste are used together, however, in many clinical trials, employed to assess mouthrinse activity, toothpaste use is suspended. This fails to measure the effect of chemical interactions which are known to occur between toothpaste ingredients and mouthrinses. The objective of this trial was to develop a methodology which would assess the adjunctive chemical plaque inhibitory action of mouthrinses, when used with toothpaste but without the indeterminate variable of toothbrushing. The study was a single blind, randomised, 7-way crossover design, based on a variation of a 4 day plaque regrowth model. The 2 x daily rinsing regimens produced increasing plaque scores in the following order: (1) water/chlorhexidine, (2) chlorhexidine/water, (3) chlorhexidine/toothpaste slurry, (4) toothpaste slurry/chlorhexidine, (5) water/toothpaste slurry, (6) toothpaste slurry/water, (7) water/water. Chlorhexidine and water or chlorhexidine and toothpaste slurry combinations produced significantly lower plaque scores than water alone. Slurry and water combinations resulted in less plaque than water alone, but differences were not significant. Toothpaste slurry and chlorhexidine produced significantly increased plaque scores compared to chlorhexidine and water. The study suggests that, outside the Hawthorne effect, chlorhexidine rinses would be less effective in reducing plaque when used with toothpaste than when used alone. The methodology could be employed as a screening tool for the evaluation of mouthrinses expected to be used as adjuncts to normal oral hygiene methods. The same could be used to optimise oral hygiene regimens which include the use of mouthrinses. PMID- 9350558 TI - On the dynamics of periodontal tissue formation in degree III furcation defects. An experimental study in dogs. AB - The aim of the experiment was to describe the formation of periodontal tissues in degree III furcation defects following GTR therapy. The study was performed in 8 foxhound dogs. The 2nd and 4th premolars in both sides of the mandible were extracted. Furcation defects were produced in the 3rd mandibular premolars. 3 weeks later, reconstructive surgery was performed. The dogs were scheduled for sacrifice 2, 4, 8, and 20 weeks after GTR therapy. Tissue blocks containing the experimental teeth were excised, demineralized in EDTA and embedded in paraffin. Serial sections were cut in the mesio-distal plane and parallel with the long axis of the roots. The microtome was set at 7 microns. The sections were stained in hematoxyline and eosin. From each biopsy, 3 sections representing the central part of the furcation, were selected for light microscopic examination. In the healed furcation sites, descriptive histological analysis of the newly-formed tissues was performed and the relative proportions of the hard and soft tissues were determined. It was demonstrated that at 2 weeks, the furcation defect contained granulation tissue and cell-rich connective tissue, while at 4 weeks the furcation was mainly occupied by connective tissue. At 8 weeks, woven bone occupied the central portion of the furcation, whereas connective tissue and cementum were observed in the lateral portions. The furcation area at 20 weeks was comprised of newly-formed cementum, periodontal ligament and bone. The onset of cementum formation had started as early as 2 weeks after GTR therapy. The cementum formation apparently occurred in 3 phases: organisation of collagen fibers adjacent and perpendicular to the root surface (phase 1), assembly of the collagen fibers and deposition of matrix (phase 2), and addition of cells and collagen fibers organised parallel to the root surface (phase 3). Bone formation took place through a process that included (1) organisation of a fibrous connective tissue, (2) differentiation of this tissue into woven bone and, (3) maturation of the woven bone into lamellar bone and bone marrow. PMID- 9350559 TI - Clinical comparison of resorbable and non-resorbable barriers for guided periodontal tissue regeneration. AB - The purpose of this study was to compare the clinical results of guided periodontal tissue regeneration (GPTR) using a resorbable barrier manufactured from a copolymer of polylactic and polyglycolic acids (Resolut Regenerative Material) with those of non-resorbable e-PTFE barrier (Gore-Tex Periodontal Material). 12 subjects participated, 6 with similarly paired class II furcations and 6 with 2 similar 2, 3-wall periodontal lesions. The resorbable and non resorbable barriers were randomly assigned to 1 defect in each subject. Non resorbable barriers were removed in six weeks. Plaque index (PlI), gingival index (GI), probing depth (PD), clinical attachment level (CAL) and gingival recession (R) were recorded at baseline, (i.e., immediately prior to surgery) and at 12 months postsurgically. The clinical healing was similar and uneventful in both groups. Intrabony pockets depicted significant changes from baseline (p < 0.05) for probing depth reduction and gain in clinical attachment levels. No differences were found between treatments. Class II furcations showed significant improvements from baseline (p < or = 0.05) for probing depth reduction and clinical attachment gain. No differences were detected between treatments. It is concluded that the resorbable barrier tested is as effective as the nonresorbable e-PTFE barrier for the treatment of class II furcations and intrabony defects. PMID- 9350560 TI - Effects of tetracycline hydrochloride and chlorhexidine gluconate on Candida albicans. An in vitro study. AB - This study examined the effects of tetracycline hydrochloride (TCN) and chlorhexidine gluconate (CHX) on the growth and viability of Candida albicans. Subcultures of Candida albicans on Sabouraud's agar, were divided into 5 treatment groups: group 1, untreated control; group 2, 0.12% CHX; group 3, 3.0 mg/ml TCN adjusted to pH 4.5; groups 4 and 5, sodium azide free Tris buffer adjusted to pH 4.5 and pH 7.4, respectively. All groups were incubated for 10 days, and sampled and subcultured daily to determine the viability of each group. Additional samples from group 2 (day 4), group 4 (day 7) and all groups at day 10 were selected for SEM and TEM examination. Visual, SEM and TEM results showed that for groups 1, 3, 4, and 5 there was a heavy and constant uniform growth of Candida albicans throughout the period of the study. However, group 2 (CHX), showed decreasing viability and attachment from day 3 to day 10, with SEM and TEM revealing decreased blastospores and profound changes in the ultrastructural morphology, indicating inhibition of normal cell growth and replication. These results show that TCN even when used at high concentrations, in vitro, will allow uninhibited growth of Candida albicans whereas CHX inhibits cell growth and replication. PMID- 9350561 TI - Subgingival temperature in smokers and non-smokers with periodontal disease. AB - The purpose of this study was to compare subgingival temperature in a group of smokers to that of a group of non-smokers with similar levels of periodontal disease. 40 adult subjects, 20 cigarette smokers and 20 non-smokers with evidence of adult periodontitis were examined. Subgingival temperature was measured at 6 sites around each of 4 maxillary anterior teeth. Probing depth, and the presence or absence of bleeding was also recorded. In addition, the sublingual temperature was recorded. All sites were classified as diseased or healthy. Healthy sites did not bleed and had a probing depth of < or = 4 mm, diseased sites were any site which had a probing depth > or = 5 mm, or which bled on probing. Mean sublingual and site temperatures were calculated for smokers and non-smokers. Mean temperature differentials (delta T) between the sublingual temperature and the site temperature were calculated for each site. Smokers had a warmer mean sublingual temperature than non-smokers. A significant difference in subgingival site temperature was demonstrated between the smokers and non-smokers, with the mean site temperature being 0.4 degree C warmer in smokers (p < 0.01). When healthy or diseased sites were compared between smokers and non-smokers, smokers also had warmer mean site temperatures than non-smokers for both healthy and diseased sites (p < 0.01). When the mean temperature differentials (delta T) between healthy and diseased sites were compared across each group, significant differences were also found. For healthy sites, the smokers had a mean delta T 0.2 degree C lower (p < 0.01) than the non-smokers, representing warmer sites. In diseased sites however, delta T was 0.3 degree C higher (p < 0.01) in smokers, representing cooler sites. PMID- 9350562 TI - Clinical and microbiological features of subjects with adult periodontitis who responded poorly to scaling and root planing. AB - In a previous report, it was shown that scaling and root planing (SRP) decreased mean pocket depth and attachment level in subjects with adult periodontitis, as well as the levels and prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. However, a subset of subjects in that study exhibited mean loss of attachment following SRP. The purpose of the present investigation was to seek clinical and microbiological differences between subjects who responded well or poorly to SRP. 57 subjects with adult periodontitis were treated by full-mouth SRP under local anaesthetic. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth prior to and 3 months post-SRP. Attachment level measurements were repeated at each visit and differences in means between visits used to assess change. 18 subjects showed mean attachment loss 3 months post-SRP (poor response group), while 39 showed mean attachment level gain (good response group). The prevalence and levels of 40 subgingival taxa in subgingival plaque samples from the mesiobuccal site of each tooth (maximum 28 sites) in each subject prior to and 3 months post-SRP were assessed using checker-board DNA-DNA hybridization. The prevalence of each species was computed for each subject and averaged across subjects in the 2 treatment-response groups at each visit. Differences between groups were sought using the Mann-Whitney test. There were no statistically significant differences between the 2 response groups in any clinical parameter prior to therapy. Subjects in the good response group showed more attachment level gain at sites with baseline pocket depths of < 4 mm, 4-6 and > 6 mm than poor response subjects. Of 40 species evaluated, A. naeslundii genospecies 2 (A. viscosus), T. denticola, C. gracilis and C. rectus were significantly higher and more prevalent pre-therapy in the good response subjects. Mean attachment level change post SRP could be predicted using multiple linear regression with A. naeslundii genospecies 2 (A. viscosus) and T. denticola as the predictor variables (r2 = 0.373, p < 0.00001). Sites that gained > or = 2 mm of attachment post therapy showed a significant decrease in the counts of P. gingivalis (7.5 +/- 3.5 to 0.2 +/- 0.2 x 10(5)), T. denticola (8.2 +/- 3.5 to 1.8 +/- 1.1 x 10(5)) and B. forsythus (11.1 +/- 5.7 to 0.3 +/- 0.2 x 10(5)). The data of the present investigation indicate that SRP is most effective in subjects and sites with high levels of the subgingival species that this therapy affects. PMID- 9350563 TI - Short-term effects of triclosan on healing following subgingival scaling. AB - The present clinical trial was performed to evaluate short-term effects of a triclosan-containing dentifrice/gel combination on soft tissue healing, when applied supra-/sub-gingivally at periodontal sites treated with scaling and root planing. 16 subjects with moderate periodontitis participated in a 2x 2-week, split-mouth designed clinical trial. 2 combinations of gel/dentifrice (the test combination containing triclosan) were used. 2 pairs of contralateral sites with probing pocket depth (PPD) > or 5 mm, and which bled on probing (BoP +) were selected in each patient as experimental units. A baseline examination included assessments of PPD, BoP, gingival index scores, plaque index scores, and the composition of the subgingival microbiota (dark-field microscopy). The assigned quadrant was anaesthetized and the teeth exposed to meticulous scaling and root planing. Immediately after the completion of mechanical therapy, either the test or control gel was applied subgingivally at the experimental sites. The volunteer was instructed to brush his/her teeth with an assigned dentifrice and to apply the gel (via a custom-made stent) supra-gingivally 2x daily for the following 2 weeks. He/she was recalled on day 7 for a second professional subgingival gel application. Re-examinations were carried out on days 2, 7 and 14 after treatment. 1-week wash-out periods separated the 2 experimental periods. The mean PPD reductions (between days 0 and 14) were 1.8 mm and 1.9 mm for the test and control gel/dentifrice sites. The reduction in BoP and gingival index scores was significantly greater during the test than during the control regimen. No significant differences were observed between the 2 regimens regarding plaque scores and composition of the subgingival microbiota. The findings from the present investigation demonstrated that triclosan, applied both sub- and supra gingivally reduced soft tissue inflammation following scaling and root planing. PMID- 9350564 TI - Power Doppler imaging in preoperative planning and postoperative monitoring of muscle flaps. AB - PURPOSE: We assessed the utility of power Doppler imaging (PDI) in preoperative planning and postoperative evaluation of microvascular tissue transfers. METHODS: Twenty-five PDI studies were performed on 23 patients using a 5-10-MHz linear array transducer. Thirteen patients were assessed preoperatively for patency of the desired donor vessel; 8 of them had surgical scars overlying the desired vascular territory. Twelve patients (including 2 from the first group) were evaluated postoperatively for patency of the vascular anastomoses and adequacy of the blood supply to the transferred tissue. RESULTS: Twelve of the 13 patients assessed preoperatively had successful flap transfers. Four of the 8 patients with scars over the desired vascular territories had absent or aberrant arteries, necessitating a change in the operative plan. None of these patients had operative complications. Eight of the 12 patients scanned postoperatively had patent anastomoses. In 2 of these patients, impending surgery was averted when the adequacy of the tissue blood supply was established with PDI. In 4 patients, PDI showed arterial or venous compromise, which was confirmed at surgery. CONCLUSIONS: PDI is a useful technique in microsurgical tissue transfer for assessing the patency of desired donor vessels preoperatively and for postoperative evaluation of blood supply. PMID- 9350565 TI - Gray-scale and color flow sonography of pancreatic ductal adenocarcinoma. AB - PURPOSE: Current sonographic technology has enhanced imaging. This study analyzes the sonographic findings in a large series of patients with pancreatic ductal adenocarcinoma. METHODS: The sonograms of 62 patients with pathologically confirmed pancreatic ductal adenocarcinoma were retrospectively analyzed. RESULTS: Tumors were an average of 4.5 x 3.5 cm in cross section. The largest lesion was 14.0 x 9.0 cm, and the smallest was 1.8 x 1.1 cm. Forty-three tumors (69%) were located in the head of the pancreas, 1 (2%) at the junction of the head and body, and 16 (26%) in the body or tail; 2 lesions (3%) were diffuse. Tumors were ovoid or spherical in 37 patients (60%) and irregular in 25 (40%). Forty tumors (65%) markedly deformed the shape of the gland. Six lesions (10%) caused no glandular contour abnormality and were visualized only because tumor echogenicity differed from that of the normal pancreas. Thirty-four tumors (55%) were homogeneously hypoechoic compared with the normal pancreas, 2 (3%) were homogeneously hyperechoic, 1 (2%) was isoechoic, and 25 (40%) had heterogeneous echotextures. Many of the heterogeneous tumors were predominantly hypoechoic with areas of varied echogenicity. Calcifications were noted in 4 patients (6%) and small intratumoral cystic areas in 9 patients (15%). Postobstructive pseudocysts were found in 4 patients (6%). Color Doppler flow information was available for 19 patients; internal flow was detected in only 1 tumor (5%). Vascular occlusion was found in 3 patients and circumferential vascular encasement in 8; the tumors in these patients were unresectable. Tumors were noted to touch vessels in another 6 patients. CONCLUSIONS: Current sonographic equipment allows the demonstration of new findings in pancreatic carcinoma. Color Doppler sonography can define tumor involvement of blood vessels and potentially affect clinical staging and treatment decisions. PMID- 9350566 TI - Color Doppler imaging of paragangliomas in the neck. AB - PURPOSE: In this study, we describe the color Doppler imaging findings in carotid body tumors and vagal body tumors. METHODS: B-mode and color Doppler imaging were performed on 17 patients who had a total of 25 previously diagnosed paragangliomas (14 carotid body tumors and 11 vagal body tumors). RESULTS: Nineteen of 25 tumors were depicted. Five small vagal body tumors in the region of the nodose ganglion and 1 carotid body tumor could not be depicted. With B mode imaging, paragangliomas appeared as well-defined, solid, hypoechoic masses. With color Doppler imaging, hypervascularity with a low-resistance flow pattern was demonstrated in all but 1 of the 19 tumors. CONCLUSIONS: The use of color Doppler imaging in the workup of an ambiguous neck mass is advocated. PMID- 9350567 TI - Detection of congenital mullerian duct anomalies using three-dimensional ultrasound. AB - PURPOSE: The purpose of this study was to assess the value of 3-dimensional sonography in the diagnosis of congenital mullerian duct anomalies, which cause infertility, preterm labor, and first trimester abortion. METHODS: A prospective study was undertaken in which 40 patients with histories of repeated spontaneous abortions or infertility were first examined using conventional 2-dimensional sonography or hysterosalpingography. Three-dimensional transvaginal sonography was then performed. RESULTS: Twenty-eight women had mullerian duct abnormalities, and 12 women had normal uterine anatomy. Mullerian duct defects detected in this study were unicornuate uterus (3), bicornuate uterus (3), complete or partial septate uterus (12), arcuate uterus (9), and didelphic uterus (1). The diagnosis of mullerian duct anomalies in these patients was confirmed by laparoscopic and/or hysteroscopic examinations. Three-dimensional sonography demonstrated all congenital uterine abnormalities with a sensitivity and specificity of 100%. Separate uterus and bicornuate uterus could be correctly diagnosed using 3 dimensional sonography in 11 (92%) of 12 cases and 3 (100%) of 3 cases, respectively. These 2 abnormalities were commonly confused with each other using hysterosalpingography and conventional sonography. CONCLUSIONS: Three-dimensional sonography with image reconstruction is less expensive and less invasive than hysterosalpingography for the assessment of uterine anatomy and diagnosis of mullerian duct abnormalities. The ability to visualize both the uterine cavity and the myometrium on a 3-dimensional scan facilitates the diagnosis of uterine anomalies and enables the differentiation of septate from bicornuate uteri for preoperative surgical planning. PMID- 9350568 TI - Clinical significance of echogenic foci in fetal lungs. AB - PURPOSE: We reviewed our experience with echogenic foci in fetal lungs. METHODS: During the period January 1991 through December 1995, 16,292 patients underwent comprehensive ultrasound examinations between 16 and 42 weeks of pregnancy. Echogenic foci in the lungs were identified in 8 fetuses. All 8 underwent karyotyping, fetal echocardiography, screening for infectious agents, and follow up sonography. The neonatal outcome was obtained in each case. RESULTS: The 5 fetuses in whom echogenic foci in the lungs were the only abnormal finding all had normal outcomes. One fetus had echogenic foci identified in 1 lung and the abdomen. This fetus tested positive for cytomegalovirus, and the pregnancy was terminated. Two fetuses with echogenic foci in the lungs had associated anomalies: 1 had an omphalocele, and the other had cerebral ventriculomegaly. Both of these pregnancies were terminated. CONCLUSIONS: In our series, isolated echogenic pulmonary foci were rare findings that carried a good prognosis. When echogenic foci in the lungs are identified, careful evaluation for associated abnormalities is warranted. PMID- 9350569 TI - Femoral artery pseudoaneurysm: Doppler sonographic features predictive for spontaneous thrombosis. AB - PURPOSE: The objective of this prospective study was to evaluate whether Doppler imaging characteristics can be used to predict spontaneous thrombosis of femoral artery pseudoaneurysms (PAs). METHODS: Eleven post-cardiac catheterization PAs were monitored with color Doppler sonography. Total volume of the lesion, volume filled with free-flowing blood, length and width of the neck of the PA, and its anatomic position were evaluated. RESULTS: All of the PAs in our series underwent spontaneous thrombosis. PAs with necks 0.9 cm or longer underwent spontaneous thrombosis in 9.8 days on average, while PAs with necks shorter than 0.9 cm required an average of 52 days to thrombose. CONCLUSIONS: PAs with longer neck lengths are more likely to thrombose in a shorter period than are those with shorter necks. It may thus be worthwhile to await spontaneous resolution when the aneurysmal neck length is 0.9 cm or more. PMID- 9350570 TI - Use of Doppler sonography to differentiate AIDS-related intrathoracic lymphoma from tuberculosis. AB - We present a case of AIDS-related intrathoracic lymphoma in which the pulmonary lesions were evaluated with Doppler sonography. The presence of internal vascularity within such lesions may help to differentiate this entity from tuberculosis. PMID- 9350571 TI - Unilateral testicular microlithiasis associated with a seminoma. AB - Unilateral testicular microlithiasis is an uncommon entity that is important because of its association with malignancy. We describe a case in which the initial clinical presentation was that of metastatic cervical lymphadenopathy. Subsequent sonographic examination of the testes revealed right testicular microlithiasis and a small, hypoechoic, ill-defined mass, which proved to be a seminoma. Since testicular microlithiasis is highly associated with testicular malignancy, it cannot be considered a benign condition. Sonographic follow-up examinations are warranted in patients with testicular microlithiasis to detect the possible development of malignancy. PMID- 9350572 TI - Klinefelter's syndrome associated with testicular microlithiasis and mediastinal germ-cell neoplasm. AB - Klinefelter's syndrome is a genetic disorder of male sexual differentiation characterized by an XXY karyotype. Although considered a benign condition, it is associated with several types of malignancies, including mediastinal germ-cell neoplasm. In addition, Klinefelter's syndrome has been rarely associated with testicular microlithiasis. Whereas patients with Klinefelter's syndrome have an increased incidence of extragonadal germ-cell neoplasms, patients with testicular microlithiasis have a predisposition to testicular germ-cell neoplasms. To our knowledge, an extragonadal germ-cell neoplasm has not been previously described in association with testicular microlithiasis. We present a patient with 2 unusual conditions, both of which are independently associated with Klinefelter's syndrome: mediastinal germ-cell neoplasm and testicular microlithiasis. PMID- 9350573 TI - Fetal acrania: five new cases and review of the literature. AB - Acrania is a lethal malformation in which there is an absence of the flat skull bones covering the brain. Five new cases are described, and a review of the English-language medical literature is presented. The sonographic differential diagnosis of acrania includes anencephaly, large cephalocele, osteogenesis imperfecta, and hypophosphatasia. The diagnosis of acrania can be established sonographically even in the first trimester if a large mass of disorganized brain tissue covered only by a thin membrane is detected. PMID- 9350574 TI - Sarcoid myopathy: imaging findings. AB - Sarcoidosis is a granulomatous multisystem disorder that may uncommonly involve muscle. Muscular sarcoid may be nodular, atrophic myopathic, or acute myositic. We illustrate a case of the myopathic type of muscular sarcoid that is unusual because the abdominal wall muscles, rather than the extremity muscles, were involved. Muscular involvement by sarcoid should be considered in the differential diagnosis of focal muscle disease, especially in a patient with a known history of sarcoid. The presence of typical bilateral hilar adenopathy on a chest radiograph as well as the presence of abdominal findings (hepatosplenomegaly and retroperitoneal adenopathy) may help establish the diagnosis. Otherwise, sonographically guided biopsy may be necessary for definitive diagnosis. PMID- 9350575 TI - Wax-and-wane hydronephrosis: sonographic finding in vesicoureteral reflux. PMID- 9350576 TI - Nephrocalcinosis induced by long-term therapy with furosemide. PMID- 9350577 TI - What's in a name? Problems with the classification of hypertension in pregnancy. PMID- 9350578 TI - Implications of the linear pressure-natriuresis relationship and importance of sodium sensitivity in hypertension. AB - Although the concept of the pressure-natriuresis curve is very clear, considerable confusion concerning its importance and utility in understanding the pathophysiology of hypertension persists. We recently showed that the pressure natriuresis curve could be considered linear. In this brief review, we would like to stress the advantages of treating it as a line. Its linear approximation simplifies understanding of the sodium sensitivity of the blood pressure and mechanisms of hypertension. The blood pressure can be expressed as the sum of two components: the non-sodium-sensitive component determined by the x intercept of the pressure-natriuresis curve and the sodium sensitive one determined by the product of the reciprocal of the slope and the amount of sodium intake. Theoretically, it can be affected in two different ways to cause hypertension; either a parallel shift along the blood pressure axis toward a higher blood pressure level due to the increase in the x intercept or a decrease in the slope. The parallel shift induces non-sodium-sensitive hypertension, whereas the decrease in slope induces sodium-sensitive hypertension. Thus, the linear approximation makes the definition of the sodium sensitivity of the blood pressure very clear and, furthermore, suggests that mechanisms of hypertension can be clarified if the determinants of the x intercept and the slope of the pressure-natriuresis curve are known. A clear definition of sodium sensitivity allows us to study its importance as a marker of a greater risk of renal and cardiovascular complications. PMID- 9350579 TI - Blood pressure in women using oral contraceptives: results from the Health Survey for England 1994. AB - OBJECTIVE: To assess whether the blood pressure is higher among women who take oral contraceptives than it is among those who do not. DESIGN: A cross-sectional survey of a stratified random sample of English adults (aged > or = 16 years). SETTING: Non-institutionalized households in England during 1994. PARTICIPANTS: From this sociodemographically representative sample of English adults, 3545 premenopausal women, of whom 892 were current users of oral contraceptives, were evaluated. INTERVENTIONS: An interviewer-administered questionnaire determined details of menopausal status, use of oral contraceptives and antihypertensive agents and other sociodemographic variables. Measurements of the weight, height and blood pressure (the mean of the last two of three readings taken with a Dinamap 8100 device) were recorded. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressures adjusted for potential confounders by oral-contraceptive-user status. RESULTS: Mean blood pressures adjusted for age were significantly higher among oral contraceptive users (125/70 mmHg) than they were among non-users (123/68 mmHg, P < 0.001 both for systolic and for diastolic blood pressures). These results remained unchanged after further adjustment for the body mass index, alcohol intake, physical activity and hypertension treatment. Blood pressure differences tended to be larger among older oral contraceptive users. Oral contraceptives containing progestogen only were not associated with higher blood pressures. CONCLUSIONS: Despite the fact that most combined oral contraceptives in current use in England contain low doses of oestrogen, slightly but significantly higher blood pressures were observed among oral contraceptive users. Blood pressures should be screened before oral contraceptives are supplied and should be monitored regularly during oral contraceptive use. PMID- 9350580 TI - Can the blood pressure predict cognitive task performance in a healthy population sample? AB - OBJECTIVES: To study the relation between the blood pressure and the neurocognitive function within the full adult age range in a large population sample. DESIGN: A cross-sectional study of 936 healthy adults who were recruited from a register of family practices, stratified for age (24-81 years), sex, and occupational level, who took part in a medical and neurocognitive test program. METHODS: The blood pressure status was studied in relation to five measures of cognitive ability, including verbal memory and speed of information processing. Other vascular risk factors were treated as control variables and included smoking, alcohol intake, body mass index, and body fat distribution. The blood pressure was measured five times using an automatic recording technique (with a Dinamap 8100 device). RESULTS: After adjustment for age, sex, and educational level in a hierarchical regression analysis, we found no unequivocal association between the mean systolic and diastolic blood pressures (or any other studied vascular risk factor) and cognitive test performance both for the whole group and for the subgroup of subjects who were not being administered antihypertensive medication and whose medical history did not include cardiovascular events. Stratified analysis within four age levels revealed no age-specific associations between the blood pressure and the cognitive function. Subjects whose blood pressure was within the hypertensive range performed worse than did matched controls at letter digit copying, but not according to other cognitive measures. CONCLUSIONS: With a population-based sample unselected for blood pressure status we found no linear relationship between the actual blood pressure level and various aspects of cognitive performance. Prospective studies are needed to investigate the possibility that the systemic blood pressure load over time is associated with a decline in specific cognitive abilities. PMID- 9350581 TI - Panic disorder, anxiety and depression in resistant hypertension--a case-control study. AB - BACKGROUND: It has been suggested that panic disorder can cause or contribute to hypertension or resistance to antihypertensive drugs. OBJECTIVE: To compare the prevalences of panic disorder, panic attacks, anxiety and depression between patients with resistant hypertension and age- and sex-matched patients with non resistant hypertension. DESIGN: A case-control study of patients attending the Sheffield Hypertension Clinic, using self-completed postal questionnaires to assess panic disorder, anxiety and depression. PATIENTS CASES: With resistant hypertension were defined as patients who presently or previously had systolic blood pressure above 160 mmHg or diastolic blood pressure above 90 mmHg despite the use of three or more antihypertensive agents at full dose. For each of 136 cases, one control with non-resistant hypertension, defined as controlled to < or = 160/90 mmHg by one or two antihypertensive agents, was identified by a bias free method. Cases and controls were matched for age and sex. MAIN OUTCOME MEASURES: Lifetime and current prevalence of panic attacks, the prevalences of panic disorder, anxiety and depression by Hospital Anxiety and Depression Scale scores, and the severity and frequency of panic attacks. RESULTS: Of the resistant hypertensive patients, 33% had experienced a panic attack compared with 39% of the control non-resistant hypertensives (resistant-non-resistant -6%, 95% confidence interval -19 to +7%). Twelve per cent of the resistant patients and 14% of controls fulfilled the criteria for a current or previous diagnosis of panic disorder (resistant-non-resistant -2%, 95% confidence interval -11% to +7%). There were also no significant differences between the groups in the prevalences of current panic attacks, panic attacks rated as moderate or worse, spontaneous panic attacks and in the frequency of panic attacks. There remained no significant difference between the groups for panic attacks and panic disorder when the analysis was limited to those patients who had idiopathic hypertension. The two groups did not differ significantly in scores for anxiety and depression measured by the Hospital Anxiety and Depression Scale. CONCLUSION: We observed no differences in the prevalences of panic, anxiety and depression between patients with resistant hypertension and non-resistant controls. These factors are probably not implicated in resistance to drug treatment. However, the prevalences of panic disorder and panic attacks were remarkably high in both groups of patients attending a hospital hypertension clinic. The relationship between panic disorder and hypertension deserves further study in a general hypertensive population. PMID- 9350582 TI - Blood pressure, serum cholesterol and nutritional state in Tanzania and in the Amazon: comparison with an Italian population. AB - OBJECTIVE: To confirm that westernization of dietary habits represents a stimulus for the expression of cardiovascular risk. DESIGN: Three representative age- and sex-matched samples of general populations of three continents were compared cross-sectionally by analysis of variance. PARTICIPANTS: In total 1110 subjects aged 22-89 years, divided into three groups (370 from Tanzania and Uganda, 370 from the Amazonian region of Brazil, and 370 from northern Italy; 111 men and 259 women in each group). RESULTS: The blood pressure of Africans eating a low-salt fish and vegetable' diet was lower than those of Brazilians, whose diet was based on cereals and meat, and highly urbanized Italians. The systolic blood pressure was correlated to the body mass index for all three populations, but with age only for the Brazilians and Italians. The total cholesterol level and body mass index, both of which are low among Africans, increased progressively with increasing economic level. CONCLUSIONS: Transition from a rural to an urbanized lifestyle is accompanied by a rise in the main cardiovascular risk factors; the present data also show that environmental rather than racial factors have a crucial impact on the risk pattern of populations. PMID- 9350583 TI - Genotype-phenotype analysis of a newly discovered family with Liddle's syndrome. AB - OBJECTIVE: To investigate the clinical, biologic, and molecular abnormalities in a family with Liddle's syndrome and analyze the short- and long-term efficacies of amiloride treatment. PATIENTS: The pedigree consisted of one affected mother and four children, of whom three suffered from early-onset and moderate-to-severe hypertension. METHODS: In addition to the biochemical and hormonal measurements, genetic analysis of the carboxy terminus of the beta subunit of the epithelial sodium channel (beta ENaC) was conducted through single-strand conformation analysis and direct sequencing. The functional properties of the mutation were analyzed using the Xenopus expression system and compared with one mutation affecting the proline-rich sequence of the beta ENaC. RESULTS: Mild hypokalemia and suppressed levels of plasma renin and aldosterone were observed in all affected subjects. Administration of 10 mg/day amiloride for 2 months normalized the blood pressure and plasma potassium levels of all of the affected subjects, whereas their plasma and urinary aldosterone levels remained surprisingly low. A similar pattern was observed after 11 years of follow-up, but a fivefold increase in plasma aldosterone was observed under treatment with 20 mg/day amiloride for 2 weeks. Genetic analysis of the beta ENaC revealed a deletion of 32 nucleotides that had modified the open reading frame and introduced a stop codon at position 582. Expression of this beta 579del32 mutant caused a 3.7 +/- 0.3-fold increase in the amiloride-sensitive sodium current, without modification of the unitary properties of the channel. A similar increase was elicited by one mutation affecting the carboxy terminus of the beta ENaC. CONCLUSIONS: This new mutation leading to Liddle's syndrome highlights the importance of the carboxy terminus of the beta ENaC in the activity of the epithelial sodium channel. Small doses of amiloride are able to control the blood pressure on a long-term basis in this monogenic form of hypertension. PMID- 9350584 TI - Role of endothelium in the endothelin-1-mediated potentiation of the norepinephrine response in the aorta of hypertensive rats. AB - OBJECTIVE: To investigate the role of the endothelium in the functional interaction between endothelin-1 and norepinephrine in the contractile response of aortas from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). METHODS: Thoracic aorta rings with and without endothelium from SHR and from WKY rats were suspended in an organ bath to record the isometric tension. After an equilibration period of 120 min, the preparations with and without endothelin-1 were subjected to single and cumulative additions of norepinephrine in different experiments. To characterize the mechanisms involved in the interaction between endothelin-1 and norepinephrine, the aortic rings were pretreated with a cyclooxygenase pathway inhibitor (piroxicam, SO29548), an inhibitor of NO synthase [NG-nitro-L-arginine (NLA)], or selective endothelin receptor blockers (BQ-123 or BQ-788). In some experiments we examined the contractile responses to norepinephrine in aortas pretreated either with angiotensin II (AII) or with U46619, an agonist of prostaglandin H2-thromboxane A2 receptors. Finally, we examined the effect of the combination of calcium-entry blockade by administration of nifedipine and treatment with either endothelin-1 or U46619 on the norepinephrine reactivity. RESULTS: Administration of 3 x 10(-10) mol/l endothelin-1 potentiated the contractile response to norepinephrine in SHR aortas with endothelium, irrespective of whether they had been treated with NLA. No endothelin-1-mediated enhancement of the response to norepinephrine was observed in SHR denuded rings and in untreated and NLA-treated WKY rat aortas. All did not affect the response to norepinephrine in SHR rings with endothelium. The amplification by endothelin-1 of the response to (1-100) x 10(-9) mol/l norepinephrine was abolished by blockade of the cyclooxygenase pathway with piroxicam or SO29548. In WKY rat and SHR denuded aortas, 10(-8) mol/l U46619 potentiated the contractile responses to norepinephrine. Administration of 3 x 10(-6) mol/l BQ-123 abolished the increase in reactivity to norepinephrine evoked by endothelin-1 in intact SHR aorta, whereas 3 x 10(-6) mol/l BQ-788 failed to modify this potentiating effect. Administration of 10(-8) mol/l nifedipine inhibited the potentiation of the norepinephrine-induced contractions evoked both by endothelin-1 in SHR aortic rings with endothelium and by U46619 in SHR denuded rings. CONCLUSION: Our results show that a low concentration of endothelin-1 induced potentiation of the contractile response to norepinephrine in SHR aortas but not in WKY rat aortas. This response was endothelium-dependent. Furthermore, our study affords functional arguments that both endothelial and smooth muscle pathways are involved in the potentiating interaction. We propose that endothelin 1 stimulates the production of endothelium- and cyclooxygenase-generated vasoconstrictor factors, which in turn may serve directly as priming stimuli at the vascular smooth muscle level, to activate the Ca(2+)-signal pathway and consequently to increase locally the vascular sensitivity to norepinephrine. PMID- 9350585 TI - The elastic modulus of conductance coronary arteries from spontaneously hypertensive rats is increased. AB - OBJECTIVE: To assess the alterations of morphological and functional properties of conductance coronary and mesenteric resistance arteries in spontaneously hypertensive rats (SHR). DESIGN: The in-vitro intrinsic elastic properties of the wall material in SHR coronary arteries were determined in comparison with those of Wistar-Kyoto (WKY) rats. Mesenteric resistance arteries from rats of both strains were also studied. METHODS: Arterial segments were cannulated at both ends using an arteriograph system and subjected to pressure increments with simultaneous measurements of the wall thickness and internal diameter. The strain, stress and incremental elastic modulus (Einc) were calculated from diameter-pressure curves. RESULTS: Over the full range of pressures tested (10 160 mmHg), the internal diameters of SHR coronary arteries were not significantly different from those of WKY rat arteries, whereas we observed that SHR mesenteric resistance arteries had a significantly smaller diameter. The stress-strain curve for coronary arteries was shifted significantly to the left-hand side for the SHR group indicating more stress per unit strain, whereas the opposite was found for mesenteric resistance arteries. When Einc was determined under isobaric conditions, we found no difference between SHR and WKY rat coronary arteries, whereas this parameter was decreased significantly for SHR mesenteric resistance arteries. When Einc was estimated at the respective operating pressures, it was 1.7- to 2.8-fold greater for SHR than it was for WKY rat mesenteric resistance and coronary arteries. Moreover, the total collagen area: lumen area ratio was significantly greater for the SHR than it was for the WKY rat coronary artery wall, but this ratio was similar for mesenteric preparations from the two strains. CONCLUSION: These results show that, at a given stress or operating pressure level, the material of SHR coronary artery wall is characterized by an increase in Einc, whereas there is no increase in Einc for in mesenteric resistance arteries. This functional alteration is accompanied by an increase in the relative proportion of collagen, a component with a high elastic modulus, in the wall. In contrast, we found no change in elastic modulus and in the relative proportion of collagen for the SHR mesenteric resistance arteries. Furthermore, the present results support the hypothesis that alterations in distensibility differ among the components of the SHR vasculature. PMID- 9350586 TI - Antisense oligodeoxynucleotide complementary to platelet-derived growth factor A chain messenger RNA inhibits the arterial proliferation in spontaneously hypertensive rats without altering their blood pressures. AB - OBJECTIVE: To evaluate the effects of the antisense oligodeoxynucleotide (ODN) to platelet-derived growth factor (PDGF) A-chain messenger RNA (mRNA) on the growth of cardiovascular organs in hypertension. DESIGN: 15-Mer antisense ODN complementary to the initiation codon region of rat PDGF-A chain mRNA and non sense ODN of identical proportion but with a random order of bases relative to that of antisense ODN were synthesized with a DNA synthesizer. METHODS: We examined the effects of the antisense ODN on the growth of vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats, and on the expression of PDGF A-chain mRNA by reverse transcription and polymerase chain reaction and PDGF A-chain protein by Western blot analysis in vitro. We evaluated the distribution of 32P-labeled antisense ODN and examined the effects of the antisense ODN on the growth of cardiovascular organs in vivo. RESULTS: The antisense ODN reduced the basal DNA synthesis of VSMC from SHR significantly, but did not do so in cells from Wistar-Kyoto rats. Mutations in the antisense ODN sequence reduced the ODN-induced inhibition of DNA synthesis. Addition of serum or transforming growth factor-beta 1 increased the DNA synthesis in the SHR-derived VSMC that was inhibited by the antisense ODN. The antisense ODN inhibited the production of PDGF A-chain protein, but not of the PDGF A-chain mRNA. The injection of 32P-antisense ODN in vivo led to a greater accumulation of radioactivity in the aorta than in other organs. Infusion of antisense ODN for 28 days did not alter the systolic blood pressure appreciably in rats of either strain. However, in SHR, it reduced markedly the elevated DNA content, [3H]-thymidine uptake, and incorporation of [3H]-thymidine into aortic DNA, and suppressed the production of aortic PDGF A-chain protein. These results indicated that the PDGF A-chain is involved in the exaggerated growth of VSMC from SHR by which inhibition of the translation of PDGF A-chain mRNA to the protein with antisense ODN occurs in vitro, and that antisense ODN to PDGF A chain suppresses the exaggerated arterial proliferation in SHR without altering the high blood pressure in vivo. CONCLUSION: These results imply that inhibition of the final responsible growth factor PDGF A-chain by antisense ODN can suppress the arterial proliferation in hypertension without altering the blood pressure, suggesting that the arterial proliferation in hypertension is independent of the high blood pressure in part, and that antisense therapy could be feasible for treating hypertension. PMID- 9350587 TI - Serum hepatocyte growth factor as a possible indicator of arteriosclerosis. AB - OBJECTIVE: To investigate the possible involvement of hepatocyte growth factor in arteriosclerotic lesions, by studying the relationship between serum concentrations of hepatocyte growth factor and grades of retinal arteriosclerosis. METHODS: We measured the blood pressure, body mass index, serum concentrations of total cholesterol, high-density lipoprotein cholesterol, triglycerides, creatinine, uric acid, total protein, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, gamma-glutamyltranspeptidase, alkaline phosphatase, and hepatocyte growth factor, erythrocyte counts, hemoglobin concentration, and hematocrit levels of 112 adults. Serum concentrations of hepatocyte growth factor were measured by a specific enzyme linked immunosorbent assay. For each subject, photographs of both optic fundi were taken, and the grade of arteriosclerotic changes in the retinal arteries was evaluated according to Scheie's classification. RESULTS: Individuals with more advanced grades of arteriosclerotic changes had higher serum hepatocyte growth factor values (grade 0, 0.056 +/- 0.004 ng/ml, n = 86; grade 1, 0.132 +/- 0.026 ng/ml, n = 17, P < 0.01, versus grade 0; grade 2-3, 0.271 +/- 0.023 ng/ml, n = 9, P < 0.01, versus grades 0 and 1). The serum hepatocyte growth factor concentrations were also correlated significantly to the serum uric acid concentrations (r = 0.230, P = 0.015) and erythrocyte counts (r = 0.299, P = 0.001), but not to the systolic and diastolic blood pressures, and other physical and humoral parameters. CONCLUSIONS: Serum hepatocyte growth factor levels are thought to indicate the presence or development of arteriosclerotic lesions and may be a useful biochemical parameter for estimating the development of systemic arteriosclerosis irrespective of blood pressure levels. PMID- 9350588 TI - The importance of pulsatile components of hypertension in predicting carotid stenosis in older adults. AB - OBJECTIVE: To evaluate pulsatile components of the blood pressure as risk markers for carotid stenosis in isolated systolic hypertension. DESIGN: Duplex scans with Doppler measures of the blood flow velocity were used to diagnose carotid stenosis in 187 participants in the Systolic Hypertension in the Elderly Program and in 187 normotensive and mildly hypertensive control subjects. METHODS: The systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure, and mean arterial pressure (MAP) were selected as independent variables. A logistic regression model for carotid stenosis was used to adjust for potentially confounding risk factors. Serial models, each containing single or double blood pressure variables, were run to compare risk markers for carotid stenosis. Receiver operating characteristic curves were compared to assess the predictive value of each model. RESULTS: In the multivariate analysis, both the SBP (P = 0.005) and the pulse pressure (P < 0.001) were predictive of carotid stenosis, but the DBP and MAP were not. However, when either the SBP or the pulse pressure was included in the model, the DBP was associated negatively with carotid stenosis (P < 0.001 and P = 0.023, respectively). An increased pulse pressure and a decreased DBP were independent risk markers for carotid stenosis. Comparison of receiver operating characteristic curves indicated that the pulse pressure had superior predictive value to the SBP (P = 0.034). CONCLUSIONS: The pulse pressure is the single best predictor of carotid stenosis. There is a negative correlation between the DBP and carotid stenosis for subjects with isolated systolic hypertension, but this can be demonstrated only after one has stratified for the SBP or for the pulse pressure. Thus, the pulsatile components of the blood pressure, increased pulse pressure and decreased DBP, are the most sensitive risk markers for the diagnoses of carotid stenosis. PMID- 9350589 TI - Actions of angiotensin and lisinopril on thalamic somatosensory neurons in normotensive, non-transgenic and hypertensive, transgenic rats. AB - OBJECTIVE: To investigate the effects of angiotensin II on discharge rates of somatosensory thalamic neurons and whether these effects are altered in hypertensive transgenic rats [TGR(mREN-2)27] and by long-term treatment with the angiotensin converting enzyme inhibitor lisinopril. DESIGN AND METHODS: Three strains of rats anesthetized with urethane were used (normotensive Wistar and Sprague-Dawley rats (SDR), and [TGR(mREN-2)27]). In addition, the effects of lisinopril treatment on SDR and transgenic animals were tested. The neuronal discharge frequency and the pattern were recorded extracellularly, and their behaviors in response to angiotensin and angiotensin antagonists administered iontophoretically were analyzed. RESULTS: Angiotensin-sensitive neurons located in the ventral posteromedial and ventral posterolateral thalamic nuclei, and in the zona incerta were excited mainly by angiotensin II. The increase in the firing rates induced by administration of angiotensin II often coincided with an increase in the number of bursts of discharges. Effects induced by angiotensin II could be blocked by administration of specific antagonists (losartan, PD 123319). Long-term treatment with lisinopril reduced the neuronal responsiveness to angiotensin II in SDR significantly in comparison with that of untreated SDR controls. Lisinopril-treated SDR had a significantly lower responsiveness to angiotensin II than did hypertensive transgenic rats that had been treated with lisinopril. CONCLUSION: The results show for the first time that administration of angiotensin II induced changes in discharge rates of somatosensory neurons, and that long-term administration of lisinopril caused a significant difference between the neuronal responsiveness to angiotensin of normotensive SDR and that of hypertensive transgenic rats. PMID- 9350590 TI - Is the tissue availability of circulating insulin-like growth factor I involved in organ damage and glucose regulation in hypertension? AB - BACKGROUND: Besides its capacity to regulate organ and tissue growth, the insulin like growth factor I exerts biologic actions that resemble those of insulin. Tissue access of the factor depends on the distribution of the circulating bound factor between its binding protein 3 that remains within the intravascular space and its binding protein 1 that is able to cross the endothelium. OBJECTIVE: To investigate whether the distribution of the circulating factor between its binding proteins is altered in patients with essential hypertension and whether this is related to changes in organ damage and glucose regulation in these patients. DESIGN: The study subjects were 30 never-treated patients with essential hypertension and 27 age- and sex-matched normotensive controls. METHODS: Serum insulin-like growth factor I-binding proteins 3 and 1 and plasma insulin-like growth factor I levels were determined by specific radioimmunoassays. RESULTS: Insulin-like growth factor I levels were significantly higher in the hypertensive patients than they were in the normotensive controls. Whereas the serum level of binding protein 1 was significantly higher in hypertensives than it was in controls, we found no differences in the level of binding protein 3 between the two groups. With the upper 100% confidence limit of the normotensive population as the cut-off point, a subgroup of 16 hypertensives had an abnormally high serum level of binding protein 1. Compared with patients with normal binding protein 1 levels, patients with increased binding protein 1 levels were characterized by the following: lower fasting glucose and insulin levels, lower insulin: glucose ratios, lower triglyceride levels, higher left ventricular mass indexes, higher creatinine clearances and higher urinary albumin excretion rates. The serum binding protein 1 level was correlated inversely to the plasma insulin level for the whole group of hypertensives. CONCLUSIONS: These results show that the distribution of circulating insulin-like growth factor I between its binding proteins 1 and 3 is altered in essential hypertension. Thus, there is a subgroup (53%) of hypertensive patients with increased serum levels of insulin-like growth factor I binding protein 1. Access of the circulating factor to tissues is more easily achieved in these patients. The clinical characteristics of this subgroup of patients suggest that the tissue availability of insulin-like growth factor I is a determinant of organ damage and insulin sensitivity in essential hypertension. PMID- 9350591 TI - Basal insulin-level oscillations in normotensive individuals with genetic predisposition to essential hypertension exhibit an irregular pattern. AB - BACKGROUND: Insulin is secreted in regular pulses at intervals of 12-14 min in normal fasting subjects. An abnormal pattern has been found in subjects with non insulin-dependent diabetes mellitus (NIDDM) and in young individuals predisposed to NIDDM. It has been suggested that there might be a causal relationship between insulin-secretion abnormalities and insulin resistance. OBJECTIVE: To examine whether insulin-secretion abnormalities are also present in offspring of patients with essential hypertension. METHODS: Eleven young (aged 18-35 years) normotensive individuals each of whom had two parents with essential hypertension were compared with 10 age- and sex-matched controls each of whom had two normotensive parents. We verified that diabetes and morbid obesity were absent among the subjects and their parents. We studied basal insulin-secretion patterns during a 60 min period, glucose tolerance by administering an oral glucose tolerance test, insulin resistance by using an isoglycaemic hyperinsulinaemic clamp and basal plasma catecholamine levels. RESULTS: Autocorrelation analysis of insulin concentrations showed that the hypertension-prone subjects had a significantly reduced or irregular oscillatory pattern compared with the regular insulin-level oscillations with a period of 12-14 min in control subjects. The hypertension-prone subjects had significantly higher systolic blood pressures and tended to be insulin-resistant. CONCLUSION: This is the first evidence of early insulin-secretion abnormalities in young normotensive individuals with a genetic predisposition to essential hypertension, but with a normal glucose tolerance and without a genetic predisposition to NIDDM. Early insulin-secretion abnormalities may be the very first step towards the development of insulin resistance and an important factor initiating the hypertension in hypertension-prone individuals. PMID- 9350592 TI - Role of left ventricular hypertrophy in diastolic dysfunction in aged hypertensive patients. AB - OBJECTIVE: To evaluate the influence of left ventricular hypertrophy (LVH) on the diastolic dysfunction in older hypertensive patients. METHODS: In total 665 patients (58% men, 61% White, aged 55-80 years) with mild-to-moderate essential hypertension underwent Doppler echocardiography. Data included left ventricular dimensions, left ventricular mass index, body mass index, E- and A-wave mitral flow velocities, E:A ratio, deceleration time > 150 ms), impaired relaxation (E:A ratio < 1.0, prolonged deceleration time according to age), and restrictive physiology (E:A ratio > 2.1, deceleration time < 150 ms)]. Data were distributed according to age (50-59, 60-69, and 70-80 years). RESULTS: The overall prevalence of sex-adjusted LVH in this study was 65%. When we compared hypertensive patients with and without LVH, the E- and A-wave velocities, E:A ratio, and deceleration time were similar. Moreover, the prevalences of normal, impaired relaxation, and restrictive physiology patterns among patients with and without LVH did not differ significantly (20, 79.5, and 0.5 versus 24, 75.5, and 0.5%). When the mitral flow patterns were adjusted according to age, the impaired relaxation pattern increased further with age (to 73% during the fifth decade, 83% during the sixth decade, and 88% during the seventh decade). CONCLUSIONS: LVH is not an independent factor associated with abnormal flow patterns in hypertensive patients aged over 50 years with normal systolic contractility. The impaired relaxation is the predominant pattern of diastolic dysfunction in older hypertensive patients and increases further with aging. PMID- 9350593 TI - Combined effects of an angiotensin converting enzyme inhibitor and a calcium antagonist on renal injury. AB - BACKGROUND: Angiotensin converting enzyme inhibitors have uniformly been shown to prevent the development both of proteinuria and of glomerulosclerosis in rats with a remnant kidney. Conversely, dihydropyridine calcium antagonists (DCA) have failed to demonstrate such a benefit in spite of causing an equivalent reduction in blood pressure. OBJECTIVE: To test the hypothesis that concomitant administration of an angiotensin converting enzyme inhibitor and a DCA would lead to a smaller increase both in proteinuria and in glomerulosclerosis relative to that caused by administration of a DCA alone at similar levels of blood pressure. METHODS: Experiments were carried out using Sprague-Dawley rats that had been subjected to five-sixths renal ablation. Animals were allocated randomly to one of four groups: control (no treatment), amlodipine (A rats), benazepril (B rats), or a combination of benazepril and amlodipine (B + A rats). We implanted intraperitoneal sensors for telemetric monitoring of the animal's blood pressure. Other parameters measured at baseline included proteinuria and inulin clearance. After approximately 7 weeks all of the parameters were remeasured and animals killed for morphologic assessment of the kidney. RESULTS: The B + A rats had lower levels of proteinuria than did the rats in group A (21 +/- 12 mg/day for B + A rats versus 59 +/- 24 mg/day for A rats, P < 0.05). The degree of glomerulosclerosis in the B + A rats was also reduced markedly compared with that in A rats (12 +/- 4% for B + A rats versus 43 +/- 12% for A rats, P < 0.05). Moreover, the results on proteinuria and glomerulosclerosis of B + A rats were similar to those for B rats. These differences could not be explained totally in terms of differences in blood pressure control (144 +/- 12 mmHg in A rats versus 132 +/- 13 mmHg in B + A rats, NS). CONCLUSION: The results were consistent with the observation that a combination of benzepril and amlodipine provides additional protection against renal injury compared with that provided by amlodipine alone. The mechanism for this benefit is not known. PMID- 9350594 TI - Evidence for decreased structurally determined preglomerular resistance in the young spontaneously hypertensive rat after 4 weeks of renal denervation. AB - OBJECTIVES: To study the effects of denervation of the kidney on renal vascular resistance at maximal dilatation and renal function during the development of hypertension in the spontaneously hypertensive rat (SHR). METHODS: SHR aged 6 weeks were subjected to left renal denervation or a sham-operation (n = 18 denervated, n = 13 sham). When they were aged 10 weeks, pairs of denervated and sham-operated left kidneys were perfused with 2% dextran in Tyrode's solution and pressure-flow and pressure-glomerular filtration rate (GFR) relationships at maximal vasodilation were established. The awake mean arterial blood pressure, in vivo renal function and renal noradrenaline content were also measured. RESULTS: There were no significant differences between the pressure-flow relationships for denervated and sham-operated kidneys. However, there was a marked, parallel, shift leftwards in the pressure-GFR relationship (P < 0.001). Thus, the denervated kidneys commenced filtering at a lower threshold perfusion pressure than did the sham-operated ones. In-vivo renal plasma flow and GFR were significantly greater in the denervated left kidneys of SHR than they were in the contralateral kidneys. The noradrenaline content in denervated kidneys was 5 +/- 3% of that in innervated kidneys. The awake mean arterial pressure was 135 +/- 1 and 138 +/- 2 mmHg in the denervated and sham-operated groups respectively. CONCLUSION: Denervation of the kidney of SHR aged 6 weeks of age altered the pressure-GFR but not the pressure-flow relationship for these rats 4 weeks later. The results are compatible with there having been an increase in average preglomerular and a decrease in post-glomerular vessel lumen diameters. These changes suggest that the renal nerves affect the structural development of the renal vasculature in SHR. PMID- 9350595 TI - Electrocardiographic and signal monitoring in ischaemic heart disease: state of the art and perspective. AB - The current role of ECG and signal monitoring in the diagnosis of Ischaemic Heart Disease is outlined in relation to imaging techniques giving accurate information on myocardial anatomy and function. ECG monitoring during stress testing remains the first step non-invasive method providing pathophysiological information. Long term continuous monitoring of the ECG and of other signals (e.g. arterial blood pressure and respiration) is commonly used to control patients with suspected or ascertained IHD. Progress of technology and of signal processing methods are driving the exploitation of signal information for diagnosis, prognosis and therapy control of ischaemic patients. PMID- 9350596 TI - ECG in stress testing: child of a lesser diagnostic god? AB - When new technologies are added to the previously existing ones, the latter can be prematurely discarded and judged obsolete not only on the basis of rational scientific facts, but also on irrational trends. Old techniques, like electrocardiography, suffer from diagnostic ambiguities that can be solved by combination with a cardiac imaging technique, like stress echocardiography. ECG monitoring during all forms of stress testing can still offer surprising dividends for a better understanding of the complex physiology of coronary artery disease, a better clinical characterization of patients with microvascular angina, and may serve as an important adjunct marker to cardiac imaging techniques. PMID- 9350597 TI - Imaging-documented cardiovascular signal database for assessing methods for ischaemia analysis. AB - A new database of cardiovascular signals has recently been developed at the CNR Institute of Clinical Physiology in a study based on patients admitted to the Coronary Care Unit for suspected ischaemic heart disease (IHD), who underwent both ECG effort stress test and echo or radionuclide diagnostic imaging procedures associated with pharmacological test of myocardial ischaemia. During stress testing, in addition to 12-lead ECG, arterial blood pressure and respiration signals are measured non-invasively and recorded. Signals and representative image frames at baseline and during ischaemia are stored in the database, which is planned to include 50 cases, annotated beat by beat and archived on CD-ROM. Each case also contains resting ECG and a comprehensive patient clinical record; if possible Holter ECG and coronary arteriography frames. PMID- 9350598 TI - Rewards in practice from chrono-meta-analyses 'recycling' heart rate, ectopy, ischemia and blood pressure information. AB - Previously published average curves of heart rate and duration of ischemia in patients with coronary artery disease, studied while on placebo or on treatment with either atenolol or diltiazem, are re-analysed for the assessment of about daily (circadian) and about-weekly (circaseptan) changes in these variables and of any treatment effect on rhythm characteristics. In addition to circadians, a circaseptan pattern characterizes the duration of ischemia in all three aforementioned study stages. Both drugs decrease the duration of ischemia, atenolol, but not diltiazem, also affects the circadian amplitude and acrophase of this variable. A circaseptan pattern is also found for heart rate on placebo and on treatment with atenolol, but not with diltiazem. Both drugs lower heart rate and the circadian amplitude and 24-h standard deviation of heart rate, atenolol much more markedly than diltiazem. Circadian and circaseptan rhythm characteristics and their alterations with treatment serve to optimize treatment by timing its administration. Chronobiologic surveillance of variables that are being readily monitored as-one-goes by modern implantable devices can also serve for the validation of the effectiveness of drug and electrical therapy. Rhythm alterations, in turn, can provide the earliest warnings of an elevated disease risk and lead to an improved diagnosis. PMID- 9350599 TI - Diagnostic acceptability of FFT-based ECG data compression. AB - There are three signal domains in which ECG data compression can be performed, namely time domain, frequency domain and parameter extraction. The present paper deals with the frequency domain method of compression using fast Fourier transform. The algorithm has been tested on the third set of the CSE database library. A performance evaluation has been made using two important parameters, namely compression ratio and percent root-mean-square difference besides visual comparison. Further, in order to know the clinical acceptable quality of the reconstructed signal peak, boundary and interval measurements were made both on the reconstructed and the original signal of the same record for comparison. The visual examination reveals that most of the noise in the original signal had been filtered out during the compression. This amounts to reduction of electromyographic noise to a considerable extent. The experimental observations show that a compression ratio of 8 is feasible while ensuring clinical acceptability. PMID- 9350600 TI - Temperature distribution in expiratory speaking flow, and early detection of vocal fold pathology. AB - This paper describes an application of heat transfer fundamentals to the development and testing of an instrument with potential use for speech production analysis. The method exploits an assumed difference between the air flow patterns of individuals with healthy and breathy voices: during breathy speech production, the glottis does not close completely, and the leakage of warm air through the glottis increases the extent of the temperature field outside the oral cavity. The proposed instrument is a pipe through which the tested individual breathes out while producing a sustained vowel. The pipe wall temperature is maintained uniform at a level considerably lower than the body temperature. The temperature gradient along the pipe centreline is measured and related to the average air velocity through the glottis. The measurements compare favourably with numerical results for the temperature field inside the instrument. These findings therefore suggest that the temperature distribution outside the oral cavity could be useful in understanding changes in air flow patterns through the vocal folds. The centreline temperature chart to be used in conjunction with the instrument is reported in dimensionless terms. PMID- 9350601 TI - An European concerted action investigating the validity of perinatal mortality as an outcome indicator for the quality of antenatal and perinatal care. AB - In this paper the concepts, objectives, design, and data analysis procedures of the EuroNatal study are described. This study started in 1996 and is a concerted action including 14 countries in Europe. The EuroNatal study aims at determining the validity of national perinatal mortality rates as an outcome indicator for the quality of antenatal and perinatal care. It is based on a conceptual model describing the relationships between differences in quality of antenatal and perinatal care, maternal and infant risk factors, variation in applied definitions, reliability of registration procedures and practices, and the outcome in terms of "true" and "observed" differences in perinatal mortality. In the first part of the study data is collected at national and aggregate level; in the second part data is collected retrospectively on individual cases of perinatal mortality in a regional sample area. Analysis of the individual cases of perinatal mortality will be by means of a perinatal audit conducted by an international expert panel. The project builds upon the work done by the participants in their respective countries. By applying common research protocols, international comparability of data collection will be enhanced and will help to create a common body of knowledge in the area of perinatal epidemiology and perinatal care. Comparison between countries is likely to lead to new insights into the strengths and weaknesses of antenatal and perinatal care systems of individual countries. PMID- 9350602 TI - Predictive value of a single CTG, ultrasound and Doppler examination to diagnose acute and chronic placental insufficiency in multiple pregnancies. AB - A non-stress test, an ultrasound biometry (biparietal and abdominal diameter) and a Doppler sonography blood flow measurement (fetal descending aorta, umbilical artery and fetal middle cerebral artery) were performed in the third trimester of 130 multiple pregnancies. These three methods were compared in terms of their prognostic value for fetal growth retardation (81 from 263 children; defined as weight at birth < 10 percentile) and a pathological "fetal outcome" (76 from 263 children, defined as 5-min-Apgar < 8, umbilical artery-pH < 7.20 and transfer to neonatal intensive care unit). Fetal growth retardation could best be predicted by means of the Doppler results for all three blood vessels ("total Doppler result") (sensitivity of 75.9%). Doppler results for all three blood vessels showed the best result in predicting a pathological "fetal outcome"; the sensitivity was 60.3%. The biometric examinations with ultrasound and the non stress test produced worse results compared to Doppler sonography. Doppler velocimetry of only one blood vessel showed worse results compared to Doppler velocimetry of more than one blood vessel. Doppler sonography should be performed as a routine test for all multiple pregnancies. More intensive pregnancy surveillance is urgently recommended with pathological findings. PMID- 9350603 TI - Changes in TcPCO2 regarding pulmonary mechanics due to pneumotachometer dead space in ventilated newborns. AB - The aim of this study was to analyze the effect of added dead space on PaCO2 after application of a pneumotachometer during the measurement of pulmonary mechanics. The study was based on 24 observations of TcPCO2 changes during the measurement of pulmonary function in 9 newborns subjected to mechanical ventilation. All newborns remained stable during the 23 minutes of the test. The introduction of a low dead space pneumotachometer (1.7 mL) for 10 minutes led to an increase in TcPCO2 of 5.40 +/- 2.66 mm Hg, from 39.76 +/- 8.69 to 45.17 +/- 9.22. Pulmonary mechanics indexes that correlated with the percentage of CO2 increase were peak inspiratory flow and expiratory time/time constant. When the pneumotachometer was removed, TcPCO2 fell but remained 0.99 +/- 2.13 mm Hg above basal TcPCO2. Final TcPCO2 tended to relate negatively with the minute volume. We conclude that this transient increase in PaCO2 should be born in mind in neonates with a high basal level and can be prevented by maintaining a long expiratory time and a high minute volume. PMID- 9350604 TI - Reliability of individual umbilical artery pH measurements. AB - To examine the reliability of umbilical artery pH measurements, two study designs were employed: (1) contemporaneous measurement of two adjoining segments of umbilical cords at birth (n = 40) and (2) repeat measurements at < 5, 15, 30, 45 and 60 minutes after birth in separate sections of the same cords (n = 40). The cord sections were left at room temperature. Limits of agreement of the contemporaneous pH measurements were from -0.066 to +0.066. Mean umbilical artery blood pH declined only slightly with time. Limits of agreement for delayed measurements were -0.061 to +0.031 at 15 minutes, -0.087 to +0.033 at 30 minutes, -0.090 to +0.046 at 45 minutes and -0.091 to +0.049 at 60 minutes. We therefore conclude that in both contemporaneous and interval sampling the accuracy of umbilical artery pH measurements is subject to biological variation. The 95% confidence interval for an individual umbilical artery pH measurement typically lies between -0.066 and +0.066 of the measured value. PMID- 9350605 TI - Decreased first trimester uric acid production in future preeclamptic patients. AB - The relationship between first trimester uric acid production and later development of pregnancy induced hypertensive disorders (PIHD) was investigated. An anti-oxidant role for uric acid has been mentioned. Since uric acid and fibronectin (PF) are both markers of preeclampsia, the relationship between these two substances was also studied. Controls (n = 72) and patients with PIHD (n = 120) were selected. Uric acid was measured in serum and 24-hours urine samples (uric acid excretion) and PF in blood plasma in 270 nulliparous women at 13 +/- 2 weeks of gestation. Uric acid excretion was significantly lower in the first trimester in a group of patients who later develop PIHD as compared to patients who remain normotensive (p < 0.05), especially when corrected for body weight (p < 0.01). Patients with elevated PF levels in the first trimester showed a significantly lower uric acid excretion than patients with normal PF levels (p < 0.05). The data show diminished uric acid production in patients who will likely develop preeclampsia suggesting an impaired anti-oxidant production in the first trimester. This observation fits well with the hypothesis that an imbalance between anti-oxidant and oxidants plays an important role in the pathogenesis of preeclampsia. PMID- 9350606 TI - Plasma hypoxanthine reacts more abruptly to changes in oxygenation than base deficit and uric acid in newborn piglets. AB - Previously, high postmortem concentrations of hypoxanthine have been found in vitreous humor of children dying from sudden infant death syndrome (SIDS). We wanted to investigate further the accumulation of hypoxanthine in vitreous humor during hypoxia. Twenty-four piglets aged 9-15 days were exposed to continuous hypoxemia (180 min 11% O2, n = 6), long interval intermittent hypoxemia (60 min 11% O2, 20 min room air, n = 7) or short interval intermittent hypoxemia (10 min 9% O2, 10 min room air with (n = 6) or without (n = 5) superimposed ligation of both carotid arteries). The increase in vitreous humor Hyp was four-fold higher (p < 0.01) with ligation of the carotid arteries (14 +/- 2.4 to 38 +/- 8.9 mumol/l) than without ligation (15 +/- 2.8 to 21 +/- 5.9 mumol/l). During continuous hypoxemia, plasma Hyp (r = 0.85), Xa (r = 0.89) uric acid (UA) (r = 0.85), and base deficit (BD) (r = 0.78) increased almost linearly (p < 0.001). Plasma Hyp responded more abruptly to changes in oxygenation than base deficit (BD) and UA. Ligation of the carotid arteries had a strong impact on Hyp accumulation in vitreous humor, suggesting that vitreous humor Hyp is not merely a filtration product of plasma Hyp, but reflects local hypoxia/ischemia in the eye. PMID- 9350608 TI - Fibrinolysis changes in normal pregnancy. AB - The aim of this study was to evaluate the changes in fibrinolysis parameters during pregnancy. Normal pregnant women (n = 60) formed the study population. Blood samples were taken in the first, second and third trimester, during delivery and three days after delivery. Fibrinolysis parameters were estimated using commercial tests. Tissue plasminogen activator, D-dimer and plasminogen activator inhibitors (PAI-1 and PAI-2) were determined. Tissue plasminogen activator and D-dimer increased after the first trimester and reached maximum levels during delivery. Plasminogen activator inhibitors type 1 and type 2 were also higher, in particular PAI-2, and reached maximum levels in the third trimester. On the third day after delivery, fibrinolysis activity recovered, but D-dimer and PAI-2 levels remained above the normal non-pregnant range. PMID- 9350609 TI - Intervillous circulation in all three trimesters of normal pregnancy assessed by color Doppler. AB - Our cross-sectional study included 115 healthy pregnant women in gestational age between 7 and 24 weeks. The aim of the study was to compare resistance (RI) and pulsatility (PI) indices, peak systolic velocity (PSV), end-diastolic velocity, and temporal averaged maximum velocity (TAMV) of the spiral arteries and vessels within the intervillous space in all three trimesters of pregnancy. The impedance to blood flow within the intervillous space significantly decreased towards the mid-pregnancy (p < 0.01) and then remained stable. Blood flow velocities within the intervillous space expressed by PSV, EDV and TAMV increased significantly (p < 0.01) towards the mid-pregnancy. After reaching the plateau between 16 and 22 weeks of gestation, these parameters remained almost constant until the 36th gestational week. Near the term low-significant decrease of blood flow velocities was noted: for PSV and TAMV p < 0.07, and for EDV p < 0.05. A significant increase in continuous intervillous blood flow velocity was noted from the 11th week onward (28 +/- 12 vs. 36 +/- 12) until the 36th week of gestation (36 +/- 12 cm/s). The first report of the haemodynamic changes within the intervillous space during pregnancy may have implication in better understanding of the metabolical interchange between maternal and fetal side. PMID- 9350607 TI - Comparison of vancomycin and teicoplanin for prophylaxis of sepsis with coagulase negative staphylococci (CONS) in very low birth weight (VLBW) infants. AB - A prospective randomized study was performed to evaluate the efficacy of a low dose (5 mg/kg/day bid) vancomycin compared with a low dose (5 mg/kg/day once daily) teicoplanin therapy to prevent CONS sepsis in VLBW-infants. All infants received this therapy after their 4th day of life or after an eventual therapy of early onset sepsis as long as an i.v. line was in place or 1500 g body weight. Twenty-seven infants were treated with vancomycin (birth weight 1103 +/- 286 g, gest. age 28.8 +/- 1.9 weeks), 28 with teicoplanin (birth weight 1133 +/- 226 g, gest. age 29.04 +/- 2.2 weeks). The infants were observed for clinical and laboratory signs of sepsis. On day 4 of therapy and every 3rd day during therapy serum creatinine levels, tracheal aspirates, stool cultures and vancomycin/teicoplanin peak and trough levels were obtained. We could not detect any case of blood culture positive sepsis and 1 case of suspected sepsis (neg. blood cultures) in both groups, as compared with a former CONS sepsis rate of 24% in our institution's VLBW infants without antibiotic prophylaxis. Nine patients in the vancomycin and five in the teicoplanin group had tracheal colonization with CONS. In both groups peak and trough levels of antibiotics were in the bactericidal range. Serum creatinine was not normal in both groups. We conclude that teicoplanin is preventing CONS sepsis as well is vancomycin. The minimal inhibitory concentrations of both antibiotics against grampositive isolates in units using this strategy have to be observed carefully to detect emerging resistance. PMID- 9350610 TI - Neonatal sepsis due to echovirus 18 infection. AB - Clinical manifestations of neonatal echovirus type 18 infections include a nonspecific febrile illness, diarrhea, and meningitis with or without exanthem. We report a successful outcome in a case of neonatal sepsis with shock caused by echovirus type 18, a complication not previously associated with this serotype. PMID- 9350611 TI - Alterations of lymphocyte subsets in patients with recurrent fetal wastage positive for antiphospholipid antibodies treated with Chinese herbal medicine. AB - The alterations of peripheral blood lymphocyte subsets were sequentially analyzed in patients with recurrent fetal wastage who were treated with the Chinese herbal medicine, Sairei-to (Chan ling-Tan) for positive antiphospholipid antibodies to analyze the underlying mechanisms of the therapy. The titer of antiphospholipid antibodies was significantly decreased by administration of Sairei-to at one and two months after commencement of treatment and in the newly pregnant state compared with that before administration. The percentage of CD19-positive cells significantly decreased at two months after commencement of Sairei-to treatment and at the newly pregnant state compared with that before administration in successful pregnancies. The percentage of CD4-positive cells significantly increased two months after commencement of Sairei-to treatment compared with that before administration in both successful pregnancies and total cases. The CD4/CD8 ratio was increased significantly after two months administration of Sairei-to in both successful pregnancies and total cases. Thus, it is suggested that the administration of Sairei-to might induce the predominance of CD4-positive cells in parallel with the suppression of antiphospholipid antibodies. Moreover, the suppression of CD19-positive cells (B cells) was induced by administration of Sairei-to which might be involved in successful continuation of pregnancy in the patients. PMID- 9350612 TI - Hydrops fetalis caused by chylothorax: an exception to the rule. PMID- 9350613 TI - Ca2+ and Na+ permeability of high-threshold Ca2+ channels and their voltage dependent block by Mg2+ ions in chick sensory neurones. AB - 1. The Mg2+ block of Na+ and Ca2+ currents through high-voltage activated (HVA; L and N-type) Ca2+ channels was studied in chick dorsal root ganglion neurones. 2. In low extracellular [Ca2+] (< 10(-8) M) and with Na+o and Cs+i as the main charge carriers (120 mM), HVA Na+ currents started to activate at -40 mV, reached inward peak values near 0 mV and reversed at about +40 mV. 3. Addition of 30-500 microM Mg2+ to the bath caused a strong depression of inward Na+ currents that was voltage and dose dependent (KD = 39 microM in 120 mM Na+ at -10 mV). The block was maximal at negative potentials (< -70 mV) and decreased with increasing positive potentials, suggesting that Mg2+ cannot escape to the cell interior. 4. Block of Ca2+ currents by Mg2+ was also voltage dependent, but by three orders of magnitude less potent than with Na+ currents (KD = 24 mM in 2 mM Ca2+ at -30 mV). The high concentration of Mg2+ caused a prominent voltage shift of channel gating kinetics induced by surface charge screening effects. To compensate for this, Mg2+ block of inward Ca2+ currents was estimated from the instantaneous I-V relationships on return from very positive potentials (+100 mV). 5. Inward Na+ and Ca2+ tail currents following depolarization to +90 mV were markedly depressed, suggesting that channels cleared of Mg2+ ions during strong depolarization are quickly re-blocked on return to negative potentials. The kinetics of re-block by Mg2+ was too fast (< 100 microseconds) to be resolved by our recording apparatus. This implies a rate of entry for Mg2+ > 1.45 x 10(8) M-1 S-1 when Na+ is the permeating ion and a rate approximately 3 orders of magnitude smaller for Ca2+. 6. Mg2+ unblock of HVA Na+ currents at +100 mV was independent of the size of outward currents, whether Na+, Cs+ or NMG+ were the main internal cations. 7. Consistent with the idea of a high-affinity binding site for Ca2+ inside the channel, micromolar amounts of Ca2+ caused a strong depression of Na+ currents between -40 and 0 mV, which was effectively relieved with more positive as well as with negative potentials (KD = 0.7 microM in 120 mM Na+ at -20 mV). In this case, the kinetics of re-block could be resolved and gave rates of entry and exit for Ca2+ of 1.4 x 10(8) M-1 S-1 and 2.95 x 10(2) s-1, respectively. 8. The strong voltage dependence and weak current dependence of HVA channel block by divalent cations and the markedly different KD values of Na+ and Ca2+ current block by Mg2+ can be well described by a previously proposed model for Ca2+ channel permeation based on interactions between the permeating ion and the negative charges forming the high-affinity binding site for Ca2+ inside the pore (Lux, Carbone & Zucker, 1990). PMID- 9350614 TI - Kinetics of Ca2+ release by InsP3 in pig single aortic endothelial cells: evidence for an inhibitory role of cytosolic Ca2+ in regulating hormonally evoked Ca2+ spikes. AB - 1. The role of the InsP3 receptor and its interaction with Ca2+ in shaping endothelial Ca2+ spikes was investigated by comparing InsP3-evoked intracellular Ca2+ release with hormonally evoked Ca2+ spikes in single endothelial cells. 2. InsP3 was generated by flash photolysis of intracellular caged InsP3. InsP3 at 0.2 microM or higher released Ca2+ from stores with a time course comprising a well-defined delay, a fast rise of free [Ca2+] to a peak where net flux into the cystosol is zero, and a slow decline to preflash levels. InsP3-evoked Ca2+ flux into unit cytosolic volume was measured as the rate of change of free [Ca2+]i during the fast rise, d[Ca2+]i/dt (mol s-1 l-1). 3. The mean delay decreased from 433 ms at 0.2 microM to 30 ms at 5 microM. At very high InsP3 concentrations, 78 microM, the delay was shorter, < 10 ms. At low InsP3 concentration the delay was reduced by approximately 30% by prior elevation of free [Ca2+]i, supporting a co operative action of free [Ca2+] and InsP3 in activation. 4. Both Ca2+ flux and peak free [Ca2+]i increased with InsP3 concentration within each cell. Maximal activation was at > 5 microM, 50% maximum Ca2+ flux was at 1.6 microM InsP3 and the Hill coefficient was between 3.6 and 4.3. A large variation of Ca2+ flux and peak [Ca2+]i was found from cell to cell at the same InsP3 concentration. 5. Strong inhibition of InsP3-evoked flux was produced by an immediately preceding response, with complete inhibition at peak free [Ca2+]i due to the first pulse. InsP3 sensitivity returned over 1-2 min, with 50% recovery at approximately 25 s. The recovery of InsP3 sensitivity may determine the minimum interval between hormonally evoked spikes. 6. Ca2+ flux due to a pulse of InsP3 terminated rapidly, in the continued presence of InsP3, producing a well-defined peak [Ca2+]. A reciprocal relation was found between the duration and the rate of Ca2+ flux, such that high Ca2+ flux was of brief duration. The rate of termination of flux measured as the reciprocal of the 10-90% rise time of free [Ca2+]i showed a linear correlation with Ca2+ flux over a large range in all cells. A systematic deviation from linearity at low InsP3 concentration showed a greater rate of termination at low InsP3 concentration than at high for the same flux. 7. Elevating cytosolic free [Ca2+] by 0.1-2.5 microM strongly inhibited Ca2+ release by InsP3, and buffering free [Ca2+] to low levels greatly prolonged Ca2+ release. Both results support the idea that Ca2+ flux quickly produces locally high free [Ca2+] which inhibits the receptor and terminates Ca2+ release. 8. Hormonally evoked Ca2+ spikes showed a similar reciprocal relation between rise time and Ca2+ flux, seen in the initial Ca2+ spike evoked by extracellular ATP in porcine aortic endothelial cells and by acetylcholine in rat aortic endothelial cells in situ, supporting the idea that the same mechanism of cytosolic Ca2+ inhibition determines the duration of hormonally and InsP3-evoked Ca2+ spikes. PMID- 9350615 TI - The interaction of nucleotides with the tolbutamide block of cloned ATP-sensitive K+ channel currents expressed in Xenopus oocytes: a reinterpretation. AB - 1. We have examined the mechanism by which nucleotides modulate the tolbutamide block of the beta-cell ATP-sensitive K+ channel (KATP channel), using wild-type and mutant KATP channels heterologously expressed in Xenopus oocytes. This channel is composed of sulphonylurea receptor (SUR1) and pore-forming (Kir6.2) subunits. 2. The dose-response relation for tolbutamide block of wild-type KATP currents in the absence of nucleotide showed both a high-affinity (Ki = 2.0 microM) and a low-affinity (Ki = 1.8 mM) site. 3. The dose-response relation for tolbutamide block of Kir6.2 delta C36 (a truncated form of Kir6.2 which is expressed independently of SUR1) was best fitted with a single, low-affinity site (Ki = 1.7 mM). This indicates that the high-affinity site resides on SUR1, whereas the low-affinity site is located on Kir6.2. 4. ADP (100 microM) had a dual effect on wild-type KATP currents: the nucleotide enhanced the current in the presence of Mg2+, but was inhibitory in the absence of Mg2+. Kir6.2 delta C36 currents were blocked by 100 microM ADP in the presence of Mg2+. 5. For wild-type KATP currents, the blocking effect of 0.5 mM tolbutamide appeared greater in the presence of 100 microM MgADP (84 +/- 2%) than in its absence (59 +/- 4%). When SUR1 was mutated to abolish MgADP activation of KATP currents (K719A or K1384M), there was no difference in the extent of tolbutamide inhibition in the presence or absence of MgADP. 6. The Ki for tolbutamide interaction with either the high- or low-affinity site was unaffected by 100 microM MgADP, for both wild-type and K719A-K1384M currents. 7. MgGDP (100 microM) enhanced wild-type KATP currents and was without effect on K719A-K1384M currents. It did not affect the Ki for tolbutamide block at either the high- or low-affinity site. 8. Our results indicate that interaction of tolbutamide with the high-affinity site (on SUR1) abolishes the stimulatory action of MgADP. This unmasks the inhibitory effect of ADP and leads to an apparent increase in channel inhibition. Under physiological conditions, abolition of MgADP activation is likely to constitute the principal mechanism by which tolbutamide inhibits the KATP channel. PMID- 9350616 TI - Photodynamic triggering of calcium oscillation in the isolated rat pancreatic acini. AB - 1. Photodynamic agents, due to their photon-dependent selective activation, can selectively activate a number of physiological processes and may directly modulate signal transduction in a number of cells including pancreatic acinar cells. 2. Activation of the photodynamic agent sulphonated aluminium phthalocyanine (SALPC) triggered recurrent cytosolic calcium ([Ca2+]i) spiking in pancreatic acinar cells. 3. The photodynamically triggered calcium spiking could be blocked by phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor U73122, but not by phosphatidylcholine-specific phospholipase C inhibitor D609. 4. Removal of extracellular Ca2+ abolished spiking, as did 2 aminoethoxydiphenylborate (2-APB), an inhibitory modulator of IP3-mediated Ca2+ release from intracellular stores. 5. These data suggest that SALPC photodynamic action may permanently fix PI-PLC in an active conformation, and this produced recurrent [Ca2+]i spiking. PMID- 9350617 TI - Volume-activated DIDS-sensitive whole-cell chloride currents in trout red blood cells. AB - 1. The nystatin-perforated whole-cell recording mode of the patch-clamp technique was used to investigate the membrane conductance of trout (Oncorhynchus mykiss) red blood cells in the steady state, 5 min after exposure to hyposmotic medium and 10 min after return to normal isosmotic medium. 2. Whole-cell I-V relations showed outward rectification when red blood cells were bathed in isosmotic (320 mosmol l-1) saline solution and the patch pipette was filled with 117 mM KCl. The membrane conductance was 2.58 +/- 0.59 nS (number of experiments, n = 18) between 0 and 100 mV and 1.32 +/- 0.19 nS (n = 18) between 0 and -100 mV. Removal of Cl- from the extracellular side or incubation with the Cl- channel blocker DIDS caused a reduction in whole-cell membrane conductance by more than 50%, indicating that the membrane current was generated by Cl- ions. The remaining conductance was voltage independent and probably due to non-selective cation conductance. 3. The membrane conductance increased approximately 2-fold after cell swelling induced by exposure to hyposmotic saline solution (215 mosmol l-1). This effect was abolished in Cl(-)-free hyposmotic medium or in the presence of DIDS. 4. The return to isosmotic solution produced a fall in membrane conductance to, or below, control values. 5. We conclude that trout red blood cells possess a significant Cl- conductance in the steady state which is reversibly activated during cell swelling and contributes to volume recovery. PMID- 9350619 TI - Chloride conductance in mouse muscle is subject to post-transcriptional compensation of the functional Cl- channel 1 gene dosage. AB - 1. In mature mammalian muscle, the muscular chloride channel ClC-1 contributes about 75% of the sarcolemmal resting conductance (Gm). In mice carrying two defective alleles of the corresponding Clc1 gene, chloride conductance (GCl) is reduced to less than 10% of that of wild-type, and this causes hyperexcitability, the salient feature of the disease myotonia. Potassium conductance (GK) values in myotonic mouse muscle fibres are lowered by about 60% compared with wild-type. 2. The defective Clcadr allele causes loss of the 4.5 kb ClC-1 mRNA. Mice heterozygous for the defective Clc1adr allele contain about 50% functional mRNA in their muscles compared with homozygous wild-type mice. 3. Despite a halved functional gene dosage, heterozygous muscles display an average GCl which is not significantly different from that of homozygous wild-type animals. The GK values in heterozygotes are also indistinguishable from homozygous wild-type animals. 4. These results indicate that a regulatory mechanism acting at the post transcriptional level limits the density of ClC-1 channels. GK is probably indirectly regulated by muscle activity. PMID- 9350618 TI - Tachykinin-induced activation of non-specific cation conductance via NK3 neurokinin receptors in guinea-pig intracardiac neurones. AB - 1. Whole mount preparations from guinea-pig hearts were used to characterize the receptors and ionic mechanisms mediating the substance P (SP)-induced depolarization of parasympathetic postganglionic neurones of the cardiac ganglion. 2. Measurement of the amplitude of depolarization in response to superfusion of different tachykinin agonists (neurokinins A (NKA) and B (NKB), SP, and senktide) gave a rank-order potency of NKB = senktide > NKA > SP, indicating involvement of an NK3 receptor. The use of the selective tachykinin receptor antagonists SR 140333, SR 48986, and SR 142801 demonstrated that only the NK3 receptor antagonist SR 142801 inhibited the SP-induced depolarization. 3. The SP-induced depolarization was not inhibited by Ba2+, TEA, or niflumic acid, or altered by reduced Cl- solutions, but was attenuated in reduced Na+ solutions. Single electrode voltage clamp studies demonstrated that the SP-induced inward current increased in amplitude at more negative potentials, had a reversal potential of approximately 0 mV, and was reduced in amplitude in reduced Na+ solutions. 4. We conclude that the SP-induced depolarization in guinea-pig postganglionic parasympathetic neurones of the cardiac ganglion is due to NK3 mediated activation of a non-selective cation conductance. PMID- 9350620 TI - Stimulation-induced changes in [Ca2+] in lizard motor nerve terminals. AB - 1. Motor axons were injected ionophoretically with one of five Ca(2+)-sensitive dyes (fluo-3, Calcium Green-2, Calcium Green-5N, fluo-3FF and Oregon Green BAPTA 5N). Changes in fluorescence (delta F/Frest) within motor terminal boutons following a single action potential and brief stimulus trains were monitored with high temporal resolution using a confocal microscope. 2. Stimulation-induced increases in delta F/Frest were confined primarily to boutons, with roughly uniform increases in all the boutons of a terminal. The increase in delta F/Frest began prior to, and decayed more slowly than, the endplate potential (EPP) recorded in the underlying muscle fibre. delta F/Frest was graded with bath [Ca2+]. Both delta F/Frest and the EPP were reduced, but not eliminated, by omega conotoxin GVIA (5-10 microM). 3. For dyes with lower affinity for Ca2+ (e.g. Oregon Green BAPTA-5N, Kd approximately 60 microM) stimulation-induced increases in delta F/Frest were measured in the presence of the K+ channel blocker 3,4 diaminopyridine (3,4-DAP, 100 microM). During brief stimulus trains (4 at 50 Hz) in 3,4-DAP, the EPP exhibited profound depression, but the fluorescence increase associated with each stimulus showed little decrement, suggesting that depression was not mediated by a reduction in Ca2+ entry. 4. For dyes with a higher affinity for Ca2+ (e.g. fluo-3, Kd approximately 0.5-1 microM) stimulation-induced increases in delta F/Frest could also be measured in normal physiological saline. Increases in delta F/Frest were much greater with 3,4-DAP present, but the amplitude decreased with successive stimuli due to partial dye saturation. 5. Calculations suggested that following a single action potential the average [Ca2+] within a bouton increased by up to 150 nM in normal saline and 940 nM in 3,4-DAP. With low affinity dyes the delta F/Frest measured near the membrane had a higher peak amplitude and a faster early decay than that measured in the centre of the bouton, suggesting that substantial spatial [Ca2+] gradients exist within boutons for at least 15 ms following stimulation. PMID- 9350621 TI - Metabotropic synaptic regulation of intrinsic response properties of turtle spinal motoneurones. AB - 1. The effect of a brief train of electric stimuli in the dorsolateral funiculus on the intrinsic response properties of turtle motoneurones was investigated in transverse sections of the spinal cord in vitro. 2. Even when glutamatergic, GABAergic and glycinergic ionotropic synaptic transmission was blocked by antagonists of AMPA, NMDA, glycine and GABA receptors, dorsolateral funiculus (DLF) stimulation induced a facilitation of plateau potentials during current clamp and the underlying inward current in voltage clamp. This facilitation lasted more than 10 s. 3. The plateau potential and the facilitation by DLF stimulation was absent in the presence of 10 microM nifedipine. The DLF-induced facilitation was reduced by antagonists of 5-HT1A, group 1 metabotropic glutamate receptors and muscarine receptors. 4. These findings suggest that the intrinsic properties of spinal motoneurones are dynamically regulated by afferent synaptic activity. These afferents can be of spinal and extraspinal origin. Continuous regulation of intrinsic response properties could be a mechanism for motor flexibility. PMID- 9350622 TI - Immunoglobulins from motoneurone disease patients enhance glutamate release from rat hippocampal neurones in culture. AB - 1. The whole-cell configuration of the patch-clamp technique was used to study the effects of immunoglobulins (IgGs) from patients affected by amyotrophic lateral sclerosis (ALS) on spontaneous glutamatergic currents in rat hippocampal cells in culture. 2. Focal application of ALS IgGs (100 micrograms ml-1) to hippocampal cells induced a rise in frequency but not in amplitude of spontaneous excitatory postsynaptic currents (SEPSC) which outlasted the period of IgG application. The mean frequency ratio (ALS over control) was 3.2 +/- 0.6 (n = 19). No changes in frequency or amplitude of SEPSCs were observed after treatment with IgGs obtained from healthy donors (n = 5) or from patients with Alzheimer's disease (n = 4). 3. ALS IgGs also increased the frequency (by a factor of 2.0 +/- 0.3) but not the amplitude of miniature excitatory postsynaptic currents (mEPSC) recorded in the presence of TTX (n = 19). A rise in frequency of mEPSC was also seen in cells superfused with a calcium-free solution (n = 4). 4. In the presence of TTX, ALS IgGs did not modify the amplitude or the shape of currents evoked by AMPA (100 microM), recorded at a holding potential of -50 mV. 5. It is concluded that ALS IgGs enhance both SEPSCs and mEPSCs through a presynaptic type of action. The excessive release of glutamate from nerve endings may be the cause of motoneurone death in ALS patients. PMID- 9350623 TI - Inhibition of spontaneous EPSCs and IPSCs by presynaptic GABAB receptors on rat supraoptic magnocellular neurons. AB - 1. The function of presynaptic GABA receptors in the regulation of transmitter release in supraoptic nucleus (SON) magnocellular neurons was investigated by recording spontaneous postsynaptic currents from rat magnocellular SON neurons in a slice preparation (150 microns thick, 1.8 mm in diameter) using the whole-cell patch-clamp technique. 2. Both the spontaneous EPSCs and IPSCs were TTX resistant. The EPSCs were abolished by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), whereas the IPSCs were abolished by picrotoxin, suggesting that the EPSCs and IPSCs are synaptic inputs from glutamatergic and GABAergic neurons, respectively. 3. The selective GABAB agonist, baclofen, reduced the frequency of both the EPSCs and IPSCs without affecting the amplitude. The time constant of the decay phase of both the EPSCs and IPSCs remained unchanged after baclofen application. 4. The reduction of the frequency of the synaptic currents by baclofen was dose dependent (10 nM to 100 microM) and the EC50 values were 5.8 and 8.5 microM for the EPSCs and IPSCs, respectively. 5. The effect of baclofen (10 microM) was antagonized by the selective GABAB antagonist, 2-hydroxy-saclofen (2OH-saclofen), at 300 microM. 6. When given alone, 2OH-saclofen (100 microM) increased the frequency of both the EPSCs and IPSCs without affecting their amplitude, suggesting that endogenously released GABA in the slice acts on presynaptic GABAB receptors. 7. The GABAA agonist, muscimol, reduced the frequency of EPSCs, and picrotoxin increased the frequency of the EPSCs, suggesting that GABAA receptors also participate in the presynaptic inhibition of glutamate release. 8. Taken together, these data suggest that GABAB receptors are present on the presynaptic terminals of both GABA and glutamate neurons in the SON, and that these presynaptic GABAB receptors play an important role in the regulation of the neuronal activity in SON magnocellular neurons. PMID- 9350624 TI - cAMP-dependent reversal of opioid- and prostaglandin-mediated depression of the isolated respiratory network in newborn rats. AB - 1. Membrane potential (Vm) and resistance (Rm) of ventral respiratory group (VRG) neurons were measured in the isolated brainstem-spinal cord from newborn rats during bath application of the opioid receptor agonists fentanyl or [D-Ala2, D Leu5]-enkephalin (Ala-Leu-Enk) and of the prostaglandin E1 (PGE1). 2. PGE1 (0.1-3 microM) and fentanyl or Ala-Leu-Enk (1-50 microM) produced depression and, at higher doses, block of inspiratory nerve activity and respiration-related postsynaptic potentials. This apnoea was associated with hyperpolarization and Rm fall in 25% of thirty-two VRG neurons tested, whereas resting Vm and Rm were not changed in the other cells. 3. The selective mu- and delta-receptor blockers naloxonazine (10-20 microM) and naltrindole (50-100 microM) antagonized the effects of 5 microM fentanyl and 50 microM Ala-Leu-Enk, respectively. 4. Opioid- and PGE1-evoked respiratory depression was reversed upon elevation of endogenous cAMP levels by stimulating adenylyl cyclase with 100 microM forskolin, activating dopamine D1 receptors with 50-100 microM 6-chloro-7,8-dihydroxy-3-allyl-1-phenyl 2, 3,4,5-tetrahydro-1H-3-benzazepine (6-chloro-APB) or preventing cAMP breakdown with 50-100 microM isobutylmethylxanthine. 5. The results indicate that opioid- or prostaglandin-induced respiratory depression is due to a fall in cAMP levels in cells responsible for generation of rhythm or providing a tonic drive to the respiratory network. 6. We suggest that elevation of cAMP levels is an effective antidote in neonates against such forms of respiratory depression. PMID- 9350625 TI - Membrane currents underlying the modified electrical activity of guinea-pig ventricular myocytes exposed to hyperosmotic solution. AB - 1. Guinea-pig ventricular myocytes were superfused with hyperosmotic (sucrose) Tyrode solution (1.2-2.8 times (T) normal osmolality) for up to 40 min. Action potentials were recorded with microelectrodes, and membrane currents with the perforated- or ruptured-patch technique. 2. Hyperosmotic treatment for 20 min shrunk cell volume and hyperpolarized the membrane. Moderate (1.2-1.5 T) treatment caused biphasic changes in action potential configuration (rapid minor shortening quickly followed by lengthening to a stable 110% control duration). Severe (2.2-2.8 T) treatment caused triphasic changes (marked early shortening, strong rebound lengthening and subsequent pronounced shortening). At peak lengthening (6-10 min) action potentials (165% control duration) had a hump near 30 mV and slowed terminal repolarization. 3. In accordance with previous studies, hyperosmotic solution inhibited the delayed rectifier K+ current, and enhanced the outward Na(+)-Ca2+ exchange current (INaCa) at plateau potentials. A novel finding was that hyperosmolality reduced the amplitude of L-type Ca2+ current (ICa,L) and slowed its rate of inactivation. Experiments on myocytes loaded with indo-1 suggest that the reduction in ICa,L is due to a rapid elevation of [Ca2+]i. 4. When impaled myocytes were preloaded with EGTA, severe hyperosmotic treatment induced a rapid monotonic shortening of the action potential to a stable 20% of control duration. Addition of external K+ quickly nulled the hyperpolarization and slowly lengthened the action potential. 5. The results suggest that modified electrical activity in osmotically shrunken myocytes is primarily caused by increases in [K+]i, [Na+]i and [Ca2+]i: (i) elevated [K+]i hyperpolarizes the membrane (which may contribute to increased [Na+]i); (ii) elevated [Na+.]i shortens all phases of the action potential (increased outward directed INaCa); and (iii) elevated [Ca2+]i has antagonistic plateau shortening (inhibition of inward ICa,L) and plateau lengthening (reduced outward INaCa) influences, as well as a strong subplateau lengthening effect (enhanced inward INaCa). PMID- 9350626 TI - Fast (mainly 30-100 Hz) oscillations in the cat cerebellothalamic pathway and their synchronization with cortical potentials. AB - 1. Intracellular recordings from 216 thalamocortical (TC) neurones in the ventrolateral (VL) nucleus of intact-cortex and decorticated cats under ketamine xylazine anaesthesia revealed spontaneously occurring fast oscillations (mainly 30-100 Hz) in 86% of investigated cells. The fast depolarizing events consisted of excitatory postsynaptic potentials (EPSPs), giving rise to fast prepotentials (FPPs) in 22% of neurones, which eventually lead to full-blown action potentials. The frequency of fast events changed by factors of 2-5 in periods as short as 0.3 1.0 s. 2. The spontaneous oscillations were similar to responses evoked in VL relay neurones by stimuli to the afferent cerebellofugal axons in brachium conjunctivum (BC) and were strikingly reduced or abolished after electrolytic lesion of BC axons. 3. The amplitude and duration of fast depolarizing events were significantly reduced during the descending phase of the inhibitory postsynaptic potentials (IPSPs) in TC cells, related to spontaneous spindles or evoked by local thalamic stimulation. 4. Averaged field potentials recorded from motor cortex and triggered by EPSPs and/or action potentials of intracellularly recorded VL cells demonstrated that both spontaneous and BC-evoked fast depolarizations in VL relay neurones were coherent with fast rhythms in cortical area 4. 5. These results show that, in addition to the thalamic and cortical generation sites of the fast (so-called gamma) oscillations, prethalamic relay stations, such as deep cerebellar nuclei, are major contributors to the induction of fast rhythms which depend on the depolarization of thalamic and cortical neurones and which represent a hallmark of brain activation patterns. PMID- 9350627 TI - Electrophysiological analysis of the function of the mammalian renal peptide transporter expressed in Xenopus laevis oocytes. AB - 1. To gain information on the mode of operation of the renal proton-coupled peptide transporter PepT2, voltage clamp studies were performed in Xenopus laevis oocytes expressing the rabbit renal PepT2. 2. Using differently charged glycyl dipeptides we show that PepT2 translocates these dipeptides by an electrogenic pH dependent process that is essentially independent of the substrate net charge. The apparent substrate affinities are in the micromolar range (2-50 microM) between pH 5.5 and 7.4 and membrane potentials of +/- 0 to -50 mV. 3. Maximal substrate-evoked inward currents (Imax) are affected by membrane voltage (Vm) and extracellular pH (pHo). Potential-dependent interactions of H+/H3O+ with PepT2 seem to be mediated by a single low affinity binding site and PepT2 remains pH dependent at all voltages. 4. The effects of voltage on apparent Imax and substrate affinity display an inverse relationship. As Vm is altered from -50 to 150 mV substrate affinities decrease 10- to 50-fold whereas apparent Imax increases almost 10-fold. 5. Even at saturating H+/H3O+ and dipeptide concentrations the I-V curves did not show saturation at negative membrane potentials, suggesting that other steps in the reaction cycle and not the ligand affinity changes are rate limiting. These are possibly the conformational changes of the empty and/or loaded transporters. 6. These findings demonstrate that not only substrate affinities but also other kinetic characteristics of PepT2 differ markedly from those of the intestinal peptide transporter isoform PepT1. PMID- 9350628 TI - Hypoxia-induced catecholamine secretion in isolated newborn rat adrenal chromaffin cells is mimicked by inhibition of mitochondrial respiration. AB - 1. In newborn mammals, systemic hypoxia provokes catecholamine secretion from the adrenal medulla. In contrast to adults, this release is independent of sympathetic innervation. We have studied the cellular processes involved in hypoxia-induced catecholamine secretion, employing fluorimetric techniques to measure changes in [Ca2+]i, NADH and mitochondrial potential, and voltammetric techniques to record changes in PO2 and catecholamine secretion. 2. In adrenal chromaffin cells freshly dissociated from newborn rats, severe hypoxia increased [Ca2+]i and secretion of catecholamines, indicating that the response of the newborn adrenal medulla to hypoxia is an intrinsic property of these cells. Discrete quantal secretory events were identifiable, suggesting an exocytotic mechanism of secretion. 3. Hypoxia-induced secretion was only seen when PO2 fell below 5 mmHg, similar to the threshold arterial PO2 reported to stimulate release in vivo. Such oxygen tensions also inhibited mitochondrial metabolism, shown by an increase in NADH autofluorescence. We therefore explored the involvement of mitochondria in oxygen sensing. Inhibition of mitochondrial respiration either by CN- at complex IV or by rotenone at complex I mimicked severe hypoxia, reversibly increasing both [Ca2+]i and catecholamine secretion. The CN(-)-induced depolarization of the mitochondrial inner membrane potential preceded the increase in [Ca2+]i by approximately 6 s. 4. The effects of severe hypoxia and CN on [Ca2+]i and catecholamine secretion were not additive, suggesting a common mechanism. 5. Chemical anoxia failed to increase [Ca2+]i in a significant proportion of cells dissociated from 2- to 4-week-old rats. Thus, the sensitivity to hypoxia is specific to adrenal chromaffin cells dissociated from newborn rats. 6. These data indicate that hypoxia-induced catecholamine secretion in the newborn adrenal medulla is mediated by reversible inhibition of mitochondrial respiration, leading to an increase in [Ca2+]i and catecholamine secretion. PMID- 9350629 TI - Glucose-induced swelling in rat pancreatic beta-cells. AB - 1. Changes in relative cell volume in response to hypotonic solutions and glucose were studied in single isolated rat pancreatic beta-cells using a video-imaging technique. beta-cell electrical activity was recorded under similar conditions using the perforated patch technique. 2. Exposure of beta-cells to hypotonic solutions (10 and 33% hypotonicity) caused an immediate increase in cell volume to relative values of 1.09 and 1.33, respectively. This was followed by a gradual regulatory volume decrease. 3. Raising the concentration of glucose from 4 to 20 mM or 12 mM (with substitution of mannitol) increased beta-cell volume by 12 and 10%, respectively. This effect of glucose persisted when CO2+ was added to inhibit insulin release. Glucose-induced volume increases were sustained for the duration of exposure to elevated hexose concentration. The addition of 16 mM 3-O methylglucose, which is transported into the beta-cell but not metabolized, produced only a transient 5% increase in beta-cell volume. 4. Exposure of beta cells to a 15% hypotonic solution resulted in a transient depolarization and electrical activity. Raising the glucose concentration to 20 or 12 mM caused a sustained depolarization and generation of electrical activity. However, the addition of 16 mM 3-O-methylglucose had no effect on beta-cell membrane potential. The glucose-induced increase in volume and induction of electrical activity, when measured in single beta-cells simultaneously, showed comparable kinetics. 5. The secretion of insulin from intact pancreatic islets was stimulated by exposure to hypotonic solutions (10-33% hypotonicity). A 15% hypotonic solution stimulated insulin release to a peak value comparable to that elicited by raising the glucose concentration from 4 to 20 mM. Whereas hypotonic solutions caused a transient stimulation of insulin release, the effect of glucose was sustained. 6. It is suggested that glucose increases the volume in rat pancreatic beta-cells by a mechanism dependent upon metabolism of the sugar. The extent of cell swelling evoked by raised glucose concentrations is sufficient to depolarize the cells and induce electrical and secretory activity and may involve activation of a volume-sensitive anion conductance. PMID- 9350630 TI - Paradoxical effect of oxygen administration on breathing stability following post hyperventilation apnoea in lambs. AB - 1. Oxygen administration is thought to suppress periodic breathing (PB) by reducing carotid body activity, and yet earlier experiments in neonates have shown that PB incidence may be increased following the application of hyperoxia. To clarify this paradox, we studied the changes in the pattern of PB that occur following administration of oxygen in a lamb model of PB. 2. PB was induced in eleven of seventeen anaesthetized lambs following passive hyperventilation with air. When oxygen was administered during PB, the pattern was first enhanced, as evidenced by a sudden decrease in the ratio of the ventilatory duration to the apnoeic pause duration, and then suppressed, as evidenced by a progressive return to stable breathing which was associated with an increase in minute ventilation. 3. Five of the six lambs that did not show PB following passive hyperventilation with air could be made to do so if oxygen was substituted for air as the inspired gas following passive hyperventilation. 4. Five of the eleven lambs that showed PB following hyperventilation with air responded to the application of oxygen during PB by switching to a gross form of episodic breathing consisting of long apnoeic pauses followed by equally long periods of breathing during which minute ventilation fell progressively with time. 5. We conclude that when applied against a background of arterial hypoxaemia, oxygen has a destabilizing influence on ventilation in that (a) it accentuates the unstable breathing that occurs during PB, (b) it induces PB in lambs that exhibited stable breathing in air, and (c) it may precipitate episodic breathing. PMID- 9350631 TI - Forearm sympathetic withdrawal and vasodilatation during mental stress in humans. AB - 1. In humans, mental stress elicits vasodilatation in the muscle vascular beds of the forearm that may be neurally mediated. We sought to determine the extent to which this vasodilatation is due to sympathetic withdrawal, active neurogenic vasodilatation, or beta-adrenergically mediated vasodilatation. 2. We simultaneously measured forearm blood flow and muscle sympathetic nerve traffic to the forearm during mental stress in humans. In a second study, we measured forearm blood flow responses to mental stress after selective blockade of alpha adrenergic neurotransmission in one forearm. In a final study, we measured forearm blood flow responses to mental stress after unilateral anaesthetic blockade of the stellate ganglion, alone or in combination with selective beta adrenergic receptor blockade of the forearm. 3. During mental stress, muscle sympathetic nerve activity decreased from 5113 +/- 788 to 1509 +/- 494 total integrated activity min-1 (P < 0.05) and forearm vascular resistance decreased from 96 +/- 29 to 33 +/- 7 mmHg (dl of tissue) min ml-1 (P < 0.05). Considerable vasodilation was still elicited by mental stress after selective blockade of alpha-adrenergic neurotransmission. Vasodilatation also occurred during mental stress after stellate ganglion blockade. This dilatation was reduced by selective blockade of beta-adrenergic receptors in the forearm. 4. Our results support a role for both sympathetic withdrawal and beta-adrenergic vasodilatation as the major causes of the forearm vasodilatation during mental stress in humans. PMID- 9350632 TI - Discharge of human muscle spindle afferents innervating ankle dorsiflexors during target isometric contractions. AB - 1. There are discrepancies in the literature about the reproducibility of forces at which human muscle spindle afferents accelerate their discharge during isometric voluntary contractions. The aim of this study was to determine for single muscle spindle afferents both the reproducibility of the 'acceleration threshold' and the factors contributing to variability of 'acceleration threshold'. 2. Microneurographic recordings were made from muscle spindle afferents innervating tibialis anterior while subjects performed isometric ankle dorsiflexions. Subjects matched the force of their contractions with a visually displayed 'ramp-and-hold' template. Template parameters were determined by the force of maximal isometric ankle dorsiflexion (MVC), and expressed as per cent MVC. The required 'ramp' rate and 'hold' force was adjusted between trials (range, 0.5-5% MVCs-1 and 0.5-20% MVC, respectively). The duration of the hold phase was 4 s and, following each contraction, stretch was applied transversely to the tendon to minimize the influence of any 'after-effects' on spindle afferent responses in subsequent contractions. 3. For each contraction, the force at which the rate of muscle spindle discharge increased was defined as the 'acceleration threshold'. Of twenty-six muscle spindle afferents innervating tibialis anterior, all but two increased their discharge in the test contractions. In 90% of contractions, acceleration thresholds were less than 3.2% MVC (range, 0.01-11.9% MVC). 4. Individual muscle spindle afferents increased their discharge at similar but not identical forces in repeated contractions. There was a positive correlation between the rate of contraction and the acceleration threshold (P < 0.001), but the strength of the target contraction had no effect on the threshold, and there was no trend for thresholds to change over time. 5. The results suggest, first, that most muscle spindle endings in the human pretibial muscles receive a significant increase in fusimotor drive during relatively weak isometric efforts and secondly, that when fusimotor after-effects are controlled, much of the residual variability in 'acceleration threshold' for any one spindle in repeated contractions is due to extrafusal factors, particularly variability in contraction rate. PMID- 9350633 TI - Lymph flow dynamics in exercising human skeletal muscle as detected by scintography. AB - 1. The effects of dynamic and isometric muscle contractions on the lymph flow dynamics in human skeletal muscle were studied with a scintographic method. 2. Radioactively labelled human serum albumin (99mTc-HSA) was injected bilaterally into the vastus lateralis muscles of eight men (n = 16), four of whom had had an endurance training background. The subjects performed 100 submaximal contractions in 10 min as (i) dynamic knee extensions (CONS), (ii) isometric contractions with the knees at full extension (IMExt), or (iii) isometric contractions with knees fixed at 90 deg angle flexion (IMFlex). The exercises were separated by 65 min periods in supine rest. The level of radioactivity at the injection site was monitored by a gamma-camera, and the clearance rate of radioactivity (CR) was calculated as the fractional decrease during the periods of interest (CR unit = % min-1). 3. The clearance rate was low during the rest periods (0.04 +/- 0.05% min 1), though higher in the trained than in the sedentary subjects (0.06 +/- 0.05 vs. 0.03 +/- 0.03% min-1; P = 0.008). Exercise increased the clearance rate three to sixfold, to 0.16 +/- 0.16% min-1 during CONS, 0.20 +/- 0.15% min-1 during IMExt and 0.09 +/- 0.11% min-1 during IMFlex. There were no differences between the subject subgroups. 4. The higher clearance rate during IMExt than during IMFlex (P = 0.02) demonstrates the importance of muscle deformations on lymph propulsion and experimentally confirms the current concepts of lymph formation and propulsion in voluntarily active skeletal muscle. It is suggested that lymph propulsion by working muscle is most efficient when the muscle is able to shorten close to its minimum length. PMID- 9350634 TI - The effect of altitude hypoxia on glucose homeostasis in men. AB - 1. Exposure to altitude hypoxia elicits changes in glucose homeostasis with increases in glucose and insulin concentrations within the first few days at altitude. Both increased and unchanged hepatic glucose production (HGP) have previously been reported in response to acute altitude hypoxia. Insulin action on glucose uptake has never been investigated during altitude hypoxia. 2. In eight healthy, sea level resident men (27 +/- 1 years (mean +/- S.E.M); weight, 72 +/- 2 kg; height, 182 +/- 2 cm) hyperinsulinaemic (50 mU min-1 m-2), euglycaemic clamps were carried out at sea level, and subsequently on days 2 and 7 after a rapid passive ascent to an altitude of 4559 m. 3. Acute mountain sickness scores increased in the first days of altitude exposure, with a peak on day 2. Basal HGP did not change with the transition from sea level (2.2 +/- 0.2 mg min-1 kg-1) to altitude (2.0 +/- 0.1 and 2.1 +/- 0.2 mg min-1 kg-1, days 2 and 7, respectively). Insulin-stimulated glucose uptake rate was halved on day two compared with sea level (4.5 +/- 0.6 and 9.8 +/- 1.1 mg min-1 kg-1, respectively; P < 0.05), and was partly restored on day 7 (7.4 +/- 1.4 mg min-1 kg-1; P < 0.05 vs. day two and sea level). Concentrations of glucagon and growth hormone remained unchanged, whereas glucose, C-peptide and cortisol increased on day 2. Noradrenaline concentrations increased during the stay at altitude, while adrenaline concentrations remained unchanged. In response to insulin infusion, catecholamines increased on day 2 (noradrenaline and adrenaline) and day 7 (adrenaline), but not at sea level. 4. In conclusion, insulin action decreases markedly in response to two days of altitude hypoxia, but improves with more prolonged exposure. HGP is always unchanged. The changes in insulin action may in part be explained by the changes in counter-regulatory hormones. PMID- 9350635 TI - Depressed autoantibody synthesis in Trypanosoma cruzi-infected rats born to mothers undergoing this infection during pregnancy. AB - Earlier work indicated that Trypanosoma cruzi infection in pregnant rats decreased the amount of myocardial damage that developed in their chronically infected offspring. Given the suspected role of autoimmune mechanisms in the generation of chronic myocarditis, we evaluated whether this maternal intervention was likely to affect the synthesis of autoantibodies in infected young. Autoantibodies were investigated against molecules exhibiting cross reactivity with T. cruzi antigens or not, that is cerebroside sulphate (sulphatide) and actin, respectively. Female '1' rats (75 days old) that had been mated with syngeneic sires were separated into two groups, one challenged with living trypomastigotes at 7, 14 and 21 days following mating, and the other one given physiologic saline at the same intervals. At the time of weaning, offspring were injected with 10(6)/T. cruzi to constitute two infected groups: young born to infected mothers (InMoTc) and young delivered by uninfected mothers (CoMoTc). Serum antibodies were investigated by ELISA at 30 and 60 days post-infection, which represents acute and chronic infection, respectively. T. cruzi infection was associated with the production of anti-sulphatide antibodies, but the phenomenon was significantly less evident in InMoTc young and virtually unnoticeable during their chronic infection. Unlike the anti-sulphatide results, levels of anti-actin antibodies showed no differences between CoMoTc and InMoTc rats when compared during acute or chronic infection. The decreased production of anti-sulphatide autoantibodies of InMoTc offspring may be due to a modification of the immune repertoire of offspring because of the contact with parasite antigens during ontogeny. PMID- 9350636 TI - Chlamydia psittaci infection in sheep: a paradigm for human reproductive tract infection. AB - Chlamydiae are important reproductive tract pathogens in a wide variety of animals. In humans, chronic or repeated infection of the female genital tract with Chlamydia trachomatis has been identified as a significant factor in the development of occlusive infertility or increased risk of ectopic pregnancy. The spectrum of reproductive disease recognized in sheep to be caused by Chlamydia psittaci has been primarily restricted to pregnant animals because the organism was clearly identified as a major cause of infectious abortion. However, following pregnancy failure, a chronic chlamydial infection can become established in the reproductive tracts of experimentally infected ewes. Persistent infection of the ewe's reproductive tract may eventually result in pathology, similar to that observed in women infected with C. trachomatis, thus decreasing the breeding life of affected ewes. Furthermore, ewes that experienced C. psittaci induced abortion provide a unique opportunity to study the host: parasite dynamic as it relates to persistent infection. This natural model of persistent infection may, in some ways, be superior to more contrived models in which the chlamydial isolate is not a normal reproductive pathogen of the study animal. Thus, the study of persistent chlamydial infection in sheep may be used for the benefit of both human and veterinary medicine. PMID- 9350637 TI - The equine immune response to endometrial cups. AB - Out of all the areas of comparative immunological study in the horse, the field of reproductive immunology has proven to be one of the most fertile and exciting. Maternal immunological interactions with the fetus involve a set of events which prevent maternal rejection of trophoblastic tissue invading the uterus, and at the same time control this invasion to regulate growth and prevent damage to maternal tissues. Unique features of equine placentation make it exceptionally well-suited to studying these immunological interactions. PMID- 9350638 TI - The factor V Leiden mutation is not a common cause of recurrent miscarriage. AB - Some investigators suggest that placental thrombosis and infarction can cause recurrent miscarriage. We have shown that the common missense mutation in the factor V gene, the Leiden mutation, which renders factor Va resistant to cleavage inactivation by activated protein C, predisposes to placental thrombosis and spontaneous miscarriage. Our objective was to determine the frequency of the Leiden mutation in a population with well-characterized idiopathic recurrent miscarriage. DNA was extracted from whole blood of 40 couples with a history of idiopathic recurrent miscarriage and 25 couples with a history of proven fertility (seven or more live births). The polymerase chain reaction was used to amplify exon 10 of the factor V gene followed by allele-specific restriction with Mnl1 for mutation detection. Results were analyzed with a chi 2 contingency table. None of the 40 women with idiopathic recurrent miscarriage carried the mutation and only one of their reproductive partners was heterozygous for the mutation. Similarly, none of the control women carried the mutation, and only one of the 25 control male partners was heterozygous for the mutation. In our referral population, the factor V Leiden mutation which predisposes to thrombosis is not a common cause of recurrent miscarriage. PMID- 9350639 TI - Differential secretion of chemokines from peripheral blood in pregnant compared with non-pregnant women. AB - The maintenance of a normal pregnancy is dependent on the delicate interaction between the endocrine and the immune systems. Cytokines are thought to play a key role in pregnancy by way of local modulation of the immune system at the level of peripheral leukocytes. This study examined the potential of peripheral venous blood cultures from pregnant women throughout gestation and from non-pregnant women to produce the chemokines monocyte chemotactic protein-1 (MCP-1), interleukin-8 (IL-8) and RANTES. Significantly (P = < 0.001), higher levels of MCP-1 were released from peripheral blood cultures from pregnant women at term than during the first trimester or from women who were not pregnant. This could not be accounted for by differences in differential blood counts. Significantly higher levels (P = < 0.05) of MCP-1 were released from PBMC preparations from pregnant compared with non-pregnant women. No 'rebound' increase in MCP-1 was observed on withdrawing progesterone support to the PBMC preparations. MCP-1 was secreted predominately from CD14+ cells with those from pregnant women producing more than those from non-pregnant women. There was no statistical difference in release of IL-8 or RANTES from either peripheral blood or PBMC preparations from pregnant or non-pregnant women. IL-8 and RANTES were secreted from CD14+ and CD14 cells, respectively. The hypothesis proposed is that the monocytes are fundamentally different in pregnancy and that measurement of MCP-1 has the potential to act as a marker of pregnancy status. PMID- 9350640 TI - Hypothesis: the role of acquired tubulointerstitial disease in the pathogenesis of salt-dependent hypertension. AB - We present a new hypothesis to explain the development of salt-dependent hypertension in humans. We propose that hypertension has two phases: an early phase in which elevations in blood pressure (BP) are mainly episodic and are mediated by a hyperactive sympathetic nervous or renin-angiotensin system, and a second phase in which BP is persistently elevated and that is primarily mediated by an impaired ability of the kidney to excrete salt (NaCl). We propose that the transition from the first phase to the second occurs as a consequence of catecholamine-induced elevations in BP that preferentially damage regions of the kidney (juxtamedullary and medullary regions) that do not autoregulate well to changes in renal perfusion pressure. The catecholamine response is associated with both an increase in peritubular capillary pressure and a reduction in peritubular capillary plasma flow, resulting in injury to the peritubular capillaries with ischemia to the tubules and interstitium. The local injury triggers the release or activation (angiotensin II, adenosine, renal sympathetic nerves) or inhibition (nitric oxide, prostaglandins, dopamine) of vasoactive mediators that further augment ischemia and result in abnormal tubuloglomerular feedback and enhanced NaCl reabsorption. The peritubular capillary injury with rarefaction simultaneously blunts the pressure natriuresis mechanism. The combined effect of enhanced tubuloglomerular feedback and impaired pressure natriuresis results in a defect in NaCl excretion which, on the exposure to salt, results in the development of persistent hypertension. Evidence is provided to suggest that this may be the major mechanism for the development of salt dependent hypertension, and particularly for the hypertension associated with blacks, aging and obesity. Thus, essential hypertension may be a type of acquired tubulointerstitial renal disease. PMID- 9350641 TI - Renal magnesium handling: new insights in understanding old problems. AB - Recent research has provided new concepts in our understanding of renal magnesium handling. Although the majority of the filtered magnesium is reabsorbed within the loop of Henle, it is now recognized that the distal tubule also plays an important role in magnesium conservation. Magnesium absorption within the cTAL segment of the loop is passive and dependent on the transepithelial voltage. Magnesium transport in the DCT is active and transcellular in nature. Many of the hormonal (PTH, calcitonin, glucagon, AVP) and nonhormonal (magnesium-restriction, acid-base changes, potassium-depletion) influences that affect magnesium transport within the cTAL similarly alter magnesium absorption within the DCT. However, the cellular mechanisms are different. Actions within the loop affect either the transepithelial voltage or the paracellular permeability. Influences acting in the DCT involve changes in active transcellular transport either Mg2+ entry across the apical membrane or Mg2+ exit from the basolateral side. These transport processes are fruitful areas for future research. An additional regulatory control has recently been recognized that involves an extracellular Ca2+/Mg(2+)-sensing receptor. This receptor is present in the basolateral membrane of the TAL and DCT and modulates magnesium and calcium conservation with elevation in plasma divalent cation concentration. Further studies are warranted to determine the physiological role of the Ca2+/Mg(2+)-sensing receptor, but activating and inactivating mutations have been described that result in renal magnesium-wasting and hypermagnesemia, respectively. All of these receptor mediated controls change calcium absorption in addition to magnesium transport. Selective magnesium control is through intrinsic control of Mg2+ entry into distal tubule cells. The cellular mechanisms that intrinsically regulate magnesium transport have yet to be described. Familial diseases associated with renal magnesium-wasting provide a unique opportunity to study these intrinsic controls. Loop diuretics such as furosemide increase magnesium excretion by virtue of its effects on the transepithelial voltage thereby inhibiting passive magnesium absorption. Distally acting diuretics, like amiloride and chlorothiazide, enhance Mg2+ entry into DCT cells. Amiloride may be used as a magnesium-conserving diuretic whereas chlorothiazide may lead to potassium depletion that compromises renal magnesium absorption. Patients with Bartter's and Gitelman's syndromes, diseases of salt transport in the loop and distal tubule, respectively, are associated with disturbances in renal magnesium handling. These may provide useful lessons in understanding segmental control of magnesium reabsorption. Metabolic acidosis diminishes magnesium absorption in MDCT cells by protonation of the Mg2+ entry pathway. Metabolic alkalosis increases magnesium permeability across the cTAL paracellular pathway and stimulates Mg2+ entry into DCT cells. Again, these changes are likely due to protonation of charges along the paracellular pathway of the cTAL and the putative Mg2+ channel of the DCT. Cellular potassium-depletion diminishes the voltage-dependent magnesium absorption in the TAL and Mg2+ entry into MDCT cells. However, the relationship between potassium and magnesium balance is far from clear. For instance, magnesium-wasting is more commonly found in patients with Gitelman's disease than Bartter's but both have hypokalemia. Further studies are needed to sort out these discrepancies. Phosphate deficiency also decreases Mg2+ uptake in distal cells but it apparently does so by mechanisms other than those observed in potassium depletion. Accordingly, potassium depletion, phosphate deficiency, and metabolic acidosis may be additive. The means by which cellular potassium and phosphate alter magnesium handling are unclear. Research in the nineties has increased our understanding of renal magnesium transport and regulation, but there are many in PMID- 9350642 TI - Expression of polycystin in mouse metanephros and extra-metanephric tissues. AB - The presence of messenger RNA for the mouse homologue of the polycystic kidney disease 1 gene (PKD1) was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) methods in mouse embryo messenger RNA. A single locus for the PKD1 gene was detected on mouse chromosome 17 by fluorescent in situ hybridization. Immunoprecipitation of proteins from [35S] methionine-labeled mouse metanephric explants with an anti-polycystin antibody (Pc1) revealed high molecular weight bands, the highest being > 400 kDa. Immunoperoxidase staining of mouse embryos with Pc1 revealed expression of polycystin as early as day 8 gestation. The expression was seen in epithelial cells of the ureteric bud, in condensing blastemal cells of the developing metanephros and, subsequently, in cells of the nascent tubules. In addition, Pc1 immunoreactivity was seen in hepatocytes and biliary epithelium, cardiac and skeletal muscle, neural tissue, gut, and bronchial epithelium. In post-natal and adult mouse kidney and liver persistent slight to moderate immunoreactivity was observed. Immunofluorescent studies of cultured 13-day mouse metanephroi revealed polycystin expression in ureteric bud epithelium, early glomerular structures (that is, condensates, S shaped and comma-shaped bodies) and in proximal and distal tubular epithelia. These data indicate that the mouse has a single gene homologous to human PKD1 on chromosome 17, and polycystin is expressed in a variety of tissues during embryonic development. PMID- 9350643 TI - Immunolocalization of V1 vasopressin receptors in the rat kidney using anti receptor antibodies. AB - By using immunocytochemical techniques we have been able to localize the V1 vasopressin receptor in the rat kidney. Immunoblotting using an antiserum raised against an affinity-purified vasopressin receptor showed a 55,000 daltons protein band that has a molecular mass similar to that of the liver V1 vasopressin receptor, as demonstrated by cross-linking studies. Immunoblotting of the antibody showed a band of 55,000 daltons in A-10 cells, which contains the V1 subtype, whereas it did not stain LLC-PK1 cells, which possess the V2 subtype, showing that the antibody recognizes the V1 vasopressin receptor. The immunostaining of kidney sections with this antiserum showed a strong reaction of the connecting tubules and cortical and medullary collecting ducts. The immunostaining pattern of connecting tubule and collecting duct cells was different, that is, the former showed a staining of both the apical and basal plasma membrane but also in the cytoplasm, whereas the latter showed a strong reaction mainly in the basolateral membrane. Immunostaining of consecutive serial sections with an antiserum raised against tissue kallikrein, an enzyme present exclusively in connecting tubules, and with the anti-receptor serum allowed us to show, for the first time, the presence of the vasopressin receptor in the connecting tubule cells and their absence in intercalated cells, the other cell type present in connecting tubules. These findings support experiments carried in the eighties on the release of renal tissue kallikrein by AVP. PMID- 9350644 TI - Effect of insulin-like growth factor binding proteins on the response of proximal tubular cells to insulin-like growth factor-I. AB - The insulin-like growth factor binding proteins (IGFBP) are major modulators of insulin-like growth factor-I (IGF-I) action, but relatively little is known about their production by kidney tubular cells or about their modulating effects on the action of IGF-I on these cells. In this study we demonstrated that rabbit proximal tubular cells express the genes for IGFBP-2, -4 and -5 and secrete 24 and 32 kDa size binding proteins. The rate of IGFBP production by these cells was regulated by several growth factors including hydrocortisone, which was potently stimulatory, and EGF, which was inhibitory. The overall effect of these kidney cell-secreted IGFBPs was to inhibit the mitogenic activity of IGF-I. Similarly, recombinant IGFBP-3, the major circulating IGFBP that in kidney is produced close to the proximal tubules, also inhibited IGF-I stimulated DNA synthesis in cultured rabbit proximal tubular cells and in cultured opossum kidney (OK) cells. IGFBP-3 also inhibited basal DNA synthesis in OK cells in the absence of added IGF-I, suggesting that this IGFBP may have an IGF-I independent action. These findings highlight the important effect that IGFBPs have on the action of IGF-I on kidney cells and support the notion that the changes in IGFBPs observed in various renal diseases may contribute to the pathophysiology of these diseases. PMID- 9350646 TI - Reversibility of experimental secondary hyperparathyroidism. AB - Chronic uremia is associated with secondary hyperparathyroidism (HPT). The purpose of the present investigation was to study the reversibility of secondary HPT after reversal of uremia by an isogenic kidney transplantation in the rat. Secondary HPT was induced in two models: Model A comprised 5/6 nephrectomized rats kept on a standard diet (N = 12; PTH 210 +/- 43 pg/ml; plasma urea 24 +/- 2 mmol/liter; and normal control rats, N = 12; PTH 45 +/- 5 pg/ml; plasma urea 6 +/ 0.2 mmol/liter); and Model B comprised 5/6 nephrectomized rats kept on a high phosphorus diet (N = 12; PTH 769 +/- 157 pg/ml; plasma urea 18 +/- 2 mmol/liter). The parathyroid function was examined by measuring the secretory response of PTH to an acute induction of hypo- and hypercalcemia. Acute hypocalcemia in the hyperphosphatemic uremic rats did not significantly increase serum PTH levels (N = 6, delta Ca2+ -0.56 mmol/liter; maximal PTH 1045 +/- 164 pg/ml; basal PTH 690 +/- 134 pg/ml; NS). During hypercalcemia the PTH levels were significantly higher than in the normal controls (N = 6; minimal PTH 24 +/- 5 pg/ml vs. normal controls 5 +/- 0.2 pg/ml, P < 0.05). After 20 weeks of uremia, the uremia was reversed by the isogenic kidney transplantation. One week after reversal of the uremia the PTH levels became normal in both models A and B (28 +/- 6 and 63 +/- 16 pg/ml, respectively) and the kidney transplanted rats from model B had a normal secretory response of PTH to both hypo- and hypercalcemia. To study whether both parathyroid cell hypertrophy and hyperplasia could be down regulated, 8 uremic glands (N = 9) or 20 normal glands (N = 6) were implanted into one normal rat. Within two weeks the rats regained normocalcemia and PTH levels remained normal from the third day after the increase of glandular mass. The 20 gland rats all had normal PTH suppressibility in response to calcium (minimal PTH 5 +/- 0.3 pg/ml). In conclusion, experimental severe secondary hyperparathyroidism is reversible very quickly after the reversal of uremia. Hyperphosphatemia in uremia is important for the non-suppressibility of the parathyroid glands to calcium. In non-uremic rats even severe parathyroid hyperplasia can be controlled, resulting in normal plasma PTH and Ca2+ levels and in a normal response to hypercalcemia. Thus, the minimal PTH secretion obtained during the induction of hypercalcemia is not an expression of the parathyroid mass. PMID- 9350645 TI - Interleukin-4 ameliorates experimental glomerulonephritis and up-regulates glomerular gene expression of IL-1 decoy receptor. AB - Monocytes/macrophages and pro-inflammatory cytokines such as interleukin (IL)-1 are important in the pathogenesis of acute glomerulonephritis. The aim of this study was to examine whether IL-4, a cytokine with anti-inflammatory activity, could modulate glomerular inflammation and reduce injury in vivo. Treatment with recombinant rat IL-4 in a model of anti-glomerular basement membrane (GBM) antibody mediated glomerulonephritis in rats reduced glomerular injury. Albuminuria was less (73% less at day 4) and a lower proportion of glomeruli had capillary thrombi (79% less at day 4). In IL-4 treated rats, there was a moderate reduction in the number of macrophages in the glomeruli and also suppression of pro-inflammatory activities of the macrophages. Northern blot analysis of glomerular RNA showed that treatment with IL-4 up-regulated mRNA levels of type II IL-1 receptor (IL-1RTII). IL-1RTII, also known as IL-1 decoy receptor, may act as a decoy molecule to inhibit the effect of IL-1 beta. To our knowledge, this is the first demonstration of (i) recombinant IL-4 reducing glomerular inflammation in vivo and (ii) a treatment that increases IL-1RTII expression in association with reduction of tissue injury in vivo. PMID- 9350647 TI - Differential regulation of renal cyclooxygenase mRNA by dietary salt intake. AB - Experiments were done to investigate the influence of dietary salt intake on renal cyclooxygenase (COX) I and II mRNA levels. To this end rats were fed either a low NaCl diet (LS; 0.02% NaCl wt/wt) or a high NaCl diet (HS diet; 4% NaCl wt/wt) for 5, 10 and 20 days. After 10 days Na excretion differed 760-fold, plasma renin activity and renin mRNA were increased eight- and threefold in LS compared to HS animals. Total renal COX I mRNA decreased 50% following the LS diet and did not change after the HS diet. Conversely, COX II mRNA declined after HS intake and transiently increased after salt depletion. COX I and II mRNAs were unevenly distributed along the cortical-medullary axis with ratios of the cortex:outer medulla:papilla of 1:3:23 and 1:1:2, respectively. Cortical COX mRNAs were inversely regulated by salt intake with eightfold changes in COX II. Conversely, in medullary zones, COX I mRNA correlated directly with salt intake. We conclude that dietary salt intake influences renal cyclooxygenase mRNAs zone specifically with opposite responses between cortex and medulla. Cortical COX II mediated prostaglandin formation is probably important in low salt states whereas medullary COX I-produced prostaglandins seem to be more important for renal adaptation to a high salt intake. PMID- 9350648 TI - Overexpression of aminopeptidase A abolishes the growth promoting effects of angiotensin II in cultured mouse mesangial cells. AB - Angiotensin II (Ang II) has diverse effects on the glomerular tuft such as regulation of glomerular hemodynamics and stimulation of mesangial cell growth, and may be one pivotal factor in the progression of renal disease. In order to locally inactivate Ang II, we overexpressed aminopeptidase A (E.C. 3.4.11.7; ATA), a peptidase involved in the conversion of Ang II into angiotensin III, in a mouse mesangial cell line (MMC) that normally does not exhibit this enzyme. Stable transfections were selected in medium containing G418. ATA-overexpressing clones ATA5 and ATA21 revealed mRNA, protein, and enzyme activity in contrast to wild-type MMCs or mock-transfected Neo3 cells (stably transfected with expression vectors without ATA cDNA). There was no difference in the binding of Ang II to its putative receptors in all cell lines. Ang II increased intracellular inositol 1,4,5-triphosphate (IP3) in Neo3, but not in ATA5 and ATA21 cells. In contrast to MMCs and Neo3 cells, Ang II failed to stimulate proliferation in ATA5 and ATA21 clones as measured by [3H] thymidine incorporation and direct cell counts. However, ATA5 and ATA21 revealed a mitogenic response not different from MMCs after stimulation 2% or 10% of fetal calf serum. Treatment of ATA5 and ATA21 with 0.1 mM of the ATA-inhibitor amastatin or an ATA-inhibiting specific monoclonal antibody restored the proliferative effect of Ang II, suggesting that surface activity of ATA is involved in the attenuated mitogenesis in these cell. Our study demonstrates that it is feasible to overexpress Ang II-degrading enzymes in cultured mesangial cells and that this overexpression attenuated some effect of exogenous Ang II. These experiments are a first step toward the development of novel strategies to selectively antagonize locally generated Ang II in the kidney. PMID- 9350649 TI - Localization of bradykinin B2 binding sites in rat kidney following chronic ACE inhibitor treatment. AB - Bradykinin exerts important influences on renal hemodynamics and tubular function by acting on renal bradykinin B2 receptors. However, the precise sites and mechanisms of its actions on the kidney are not known. To help elucidate the mechanisms of renal actions of bradykinin in vivo, we have employed high resolution electron microscopic autoradiography to localize bradykinin B2 binding sites in the rat kidney following intravenous administration of a radiolabeled ligand, 125I-HPP-Hoe140 (3-4-Hydroxyphenyl-propionyl-DArg0-[Hyp3-Thi5-D-Tic 7 Oic8]-bradykinin), a derivative of the highly selective bradykinin B2 receptor antagonist, Hoe140. In non-treated rats, bradykinin B2 binding sites were localized to the cell bodies and the luminal brush border of the proximal convoluted tubules in the cortex. In the medulla (except for the outer stripe of the outer medulla), binding occurred in the distal tubules, thin limbs of the loop of Henle, collecting ducts, peritubular capillary endothelium and renomedullary interstitial cells. To exclude the possibility that the radioligand may bind to angiotensin converting enzyme, rats were pretreated with the angiotensin converting enzyme inhibitor, perindopril. In these rats, binding to the cell bodies and the luminal brush border of the proximal convoluted tubules in the cortex was completely abolished, while binding remained unaltered in the medulla. Further studies using high performance liquid chromatography revealed that while the radioligand was degraded following systemic administration in nontreated rats, the degradation was significantly reduced in the rats pretreated chronically with perindopril. These results indicate that binding detected in the proximal tubules in the normal rats is due primarily to the tubular uptake of the degraded radioligand, and that bradykinin B2 binding sites occur predominantly in the renal tubules, vascular endothelium, and renomedullary interstitial cells of the renal medulla. PMID- 9350650 TI - Effect of glycine on prelethal and postlethal increases in calpain activity in rat renal proximal tubules. AB - The effect of glycine on hypoxia- and ionomycin-induced increases in calpain activity in rat proximal tubules was determined. Calpain activity was determined both in vitro and in the intact cell using the fluorescent substrate N-succinyl Leu-Leu-Val-Tyr-7-amido-4-methyl coumarin (N-succinyl-Leu-Leu-Val-Tyr-AMC) and Western blotting for calpain-specific spectrin breakdown products (BDP), respectively. At 7.5 minutes of hypoxia (prelethal injury model) there was a significant (10-fold) increase in in vitro calpain activity that was not inhibited by glycine. At 15 minutes of hypoxia (postlethal injury model) there was a further increase in calpain activity that was inhibited by glycine. Normoxic tubules incubated with the calcium ionophore ionomycin (5 microM) for two minutes and 10 minutes had a significant increase in calpain activity that was not inhibited by glycine. After 15 minutes of hypoxia in the presence of glycine, there was an increase in calpain-specific spectrin breakdown products (BDP) in both Triton X-100 soluble and cytosolic extracts from proximal tubules. Glycine in concentrations up to 10 mM had no direct effect on the in vitro calpain activity of purified calpains. The present study demonstrates that: (1) prelethal increases in calpain activity stimulated by hypoxia and ionomycin treatment are not affected by glycine; (2) calpain-mediated spectrin breakdown during hypoxia occurs in the presence of glycine; (3) glycine does prevent the additional postlethal increase in calpain activity probably by maintaining membrane integrity to calcium influx. PMID- 9350651 TI - Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. AB - Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts, characterized by the de novo expression of alpha-smooth muscle (alpha-sm) actin and/or desmin. Both the mechanisms underlying this differentiation and their effects on cellular function are poorly understood. In vitro studies are difficult since the phenotypes of fibroblasts in culture have as yet not been well defined. We have, therefore, examined the phenotype of inner medullary fibroblasts (IMF) during the transition from in vivo to in vitro in various cell fractions derived from the inner medulla of healthy rats. IMF were positive for the lectin BSL-1 and negative for markers of endothelial cells. IMF first lost their prominent lipid droplets in vitro. Subsequently they developed cytoplasmic processes accompanied by a decrease in their reactivity for the lectin BSL-1 from strong to weak. From day 3 in primary culture, exclusively these weakly positive BSL-1 cells showed a de novo expression of alpha-sm actin (day 4 of primary culture, 75 +/- 4%; day 20, 94 +/- 2%) and desmin (day 4, 43 +/- 8%; day 20, 66 +/- 6%), classifying them as myofibroblasts. This transformation depended on culture conditions. In a mixed coculture with inner medullary collecting duct (IMCD) cells the transformation of IMF was largely absent: a significantly greater number of strong BSL-1 positive cells contained prominent lipid droplets (39 +/- 4 vs. 19 +/- 4%, P < 0.05) on day 4 of primary culture, and the transition of strongly to weakly positive BSL-1 IMF was almost completely blocked. By reducing the seeding density of IMCD cells the effect of this condition on IMF transformation could be largely abolished. This first detailed phenotypic characterization of rat fibroblasts during the transition from in vivo to in vitro demonstrates that these cells-depending on culture conditions-differentiate to myofibroblasts within a few days of primary culture and that subcultured IMF exhibit predominantly this phenotype. The presented model may serve as a useful tool for the in vitro study of myofibroblast formation and the consequences of such a differentiation for the physiological functions of IMF. PMID- 9350652 TI - Adhesion, internalization and metabolism of calcium oxalate monohydrate crystals by renal epithelial cells. AB - The interaction between crystals that nucleate in the nephron lumen and tubular cells could be an important determinant of renal calcification. Kidney epithelial cells in monolayer culture (BSC-1 line), used to model the tubule, rapidly bound and internalized crystals of calcium oxalate monohydrate (COM), the most common constituent of renal stones. Transmission and scanning electron microscopy, enzyme histochemistry, and kinetic analysis of [14C]-labeled crystals were used to study the interaction between renal cells and COM crystals. Electron microscopy revealed that adherent crystals on the apical cell surface can serve as sites for aggregation of additional crystals. Enhanced binding of exogenous crystals to plasma membrane domains overlying internalized crystals was observed for at least 24 hours after the initial cell-crystal interaction. Following internalization, crystals appeared to dissolve within lysosomal inclusion bodies during the ensuing five to seven weeks. Over this time, many cells still containing crystals clustered together in the monolayer. These observations suggest that adhesion and internalization can promote crystal retention in the nephron, whereas intracellular dissolution of crystals may serve as an important, hitherto unrecognized defense against pathologic renal calcification. PMID- 9350653 TI - Nonenzymatic glycation of type IV collagen and matrix metalloproteinase susceptibility. AB - The glomerular basement membrane (GBM) is damaged in diabetes through complex mechanisms that are not fully understood. Prominent among them is nonenzymatic protein glycation leading to the formation of so-called advanced glycation end products (AGEs). We examined the effects of in vitro glycation of intact collagen type IV in bovine lens capsule (LBM) and kidney glomerular (GBM) basement membranes on their susceptibility to matrix metalloproteinases, using stromelysin 1 (MMP-3) and gelatinase B (MMP-9). Sites of cleavage of unmodified LBM collagen were located in the triple helical region. In vitro glycation by glucose severely inhibited the release of soluble collagen cleavage peptides by MMP-3 and MMP-9. The distribution of AGEs within the three domains of collagen IV (7S, triple helical, and noncollagenous NC1) were compared for LBM glycation using AGE fluorescence, pentosidine quantitation, and immunoreactivity towards anti-AGE antibodies that recognize the AGE carboxymethyllysine (CML). Marked asymmetry was observed, with the flexible triple helical domain having the most pentosidine and fluorescent AGEs but the least CML. The in vivo relevance of these findings is supported by preliminary studies of AGE distribution in renal basement membrane (RBM) collagen IV domains from human kidneys of two insulin-dependent diabetics and one normal subject. Pentosidine and fluorescent AGE distributions of diabetic RBM were similar to LBM, but the CML AGE in diabetic kidney was less in the triple helical domain than in NC1. Our results support the hypothesis that nonenzymatic glycation of collagen IV contributes to the thickening of basement membranes, a hallmark of diabetic nephropathy. PMID- 9350654 TI - Abrogation of glomerular injury in nephrotoxic nephritis by continuous infusion of interleukin-6. AB - Interleukin-6 (IL-6) has been reported to have pro- and anti-inflammatory effects. It has also been shown to cause mesangial cell proliferation in vitro and has been suggested as a mediator of injury in proliferative nephritis. We have assessed the effects of continuous infusion of human recombinant (hr) IL-6, by osmotic minipump, on the degree of glomerular injury, and on glomerular and interstitial cell proliferation, in the accelerated autologous phase of nephrotoxic nephritis. Two groups of rats were pre-immunized with 1 mg of normal rabbit IgG in Freund's complete adjuvant. One week later, nephritis was induced by an intravenous injection of 1 ml of rabbit nephrotoxic serum. One day before the induction of nephritis, group 1 (N = 9) was subcutaneously implanted with osmotic minipumps filled with 50 micrograms (200 microliters) of IL-6 (equivalent to a dose of 6 micrograms/day), while in group 2 (N = 11) the minipumps were filled with 200 microliters of normal saline. In group 3 (N = 6) normal rats were infused with 50 micrograms of IL-6 alone. The rats were killed seven days after implantation of minipumps. The administered hrIL-6 was detectable in the circulation within the pathophysiological range, and induced a hepatic acute phase response, as assessed by alpha 2-macroglobulin levels. Continuous treatment with IL-6 resulted in a significant reduction in albuminuria (from 195 +/- 37 mg/20 hr to 60 +/- 15 mg/20 hr on day 1, and from 494 +/- 52 mg/20 hr to 238 +/- 30 mg/20 hr on day 7, P < 0.002) and in the prevalence of glomerular capillary thrombosis (from 19 +/- 3% to 5 +/- 1%, P < 0.002). There was also a reduction in macrophage infiltration (ED1 + ve cells from 524 +/- 34 to 466 +/- 14 per 50 glomeruli, P < 0.02) and activation (ED3 + ve cells from 106 +/- 13 to 42 +/- 5 per 50 glomeruli, P < 0.002). Immunohistology showed fewer interstitial Ia + ve cells (OX3 and OX4) in the IL-6 treated group. Similar results were obtained in a second set of experiments in which the IL-6 treatment was extended until day 14. Kidney sections taken from nephritic rats infused with IL-6 showed no increase in glomerular or interstitial cell proliferation when stained with antibodies to proliferating cell nuclear antigen. There was no difference in the deposition of rabbit IgG or rat IgG along the glomerular basement membrane (GBM), and the titer of rat anti-rabbit IgG was similar in the IL-6 and control treated rats. Infusion of IL-6 alone in normal rats had no functional or pathological effects. In conclusion, these results show that IL-6 has powerful anti-inflammatory effects in a rat model of anti-GBM nephritis, and does not induce mesangial cell proliferation in vivo. PMID- 9350655 TI - Isolation of proximal and distal tubule cells from human kidney by immunomagnetic separation. Technical note. AB - After collagenase digestion and Percoll density gradient centrifugation of human renal tissue, tubular epithelial cells of the proximal and the distal segments were isolated with an immunomagnetic method using MACS microbeads. To enrich proximal tubular (PT) cells we used a monoclonal antibody (mAb) against aminopeptidase M (APM, CD 13), specific of the proximal tubule. Distal tubular (DT) cells were isolated through a mAb recognizing Tamm-Horsfall glycoprotein (THG), a specific antigen for the thick ascending limb and the early distal convoluted tubule. Cells of the proximal primary isolate were histochemically strongly positive for aminopeptidase M (98.6%), however, cells of the distal portion were negative (98.7%). Ultrastructural analysis of PTC primary isolates revealed highly preserved brush border microvilli, well-developed endocytosis apparati and numerous mitochondria, whereas DTC primary isolates showed smaller cells with basolateral invaginations and less apical microvilli. Characterization by immunofluorescence indicated the coexpression of cytokeratin and vimentin, whereas staining for desmin, smooth muscle actin, a fibroblast-specific marker and von Willebrand factor was negative. Cultured PT and DT cells displayed different adenylate cyclase responsiveness to hormonal stimulation. PTH (10(-6) M) increased cAMP production in distal cells up to 32.8-fold of the basal level and in proximal only up to 3.5-fold (10(-8) M, DT 14.4x and PT 2.25x). Calcitonin stimulated adenylate cyclase in DT in a dose dependent fashion (10(-6) M, 4.3x; 10(-8) M, 2.25x), whereas only a low calcitonin response was found in PT cells (10(-6) M, 1.6x; 10(-8) M, 1.4x). AVP (10(-6) M) activated the distal cAMP production only up to 1.9x of the basal level, but the proximal cAMP-production was negligible (only 1.3x the basal level). The data of this study indicate the proximal and distal tubule origin of the cultured cells that were isolated according to their segment-specific antigens. PMID- 9350656 TI - Hyperosmotic stress up-regulates amino acid transport in vascular endothelial cells. AB - Cultured vascular endothelial cells take up L-proline by sodium-dependent transport. Cells incubated in medium made hyperosmotic by addition of sucrose showed a dose-dependent increase in Na+/proline cotransport. Studies with alpha (methylamino)isobutyric acid revealed that the up-regulation was specific for amino acid transport system A. Up-regulation was blocked by actinomycin D and cycloheximide, indicating roles for gene transcription and protein synthesis. Up regulation was maximum after five to six hours of hyperosmotic treatment, but returned to control levels when osmotic stress was maintained for 24 hours. The decline at 24 hours was accompanied by a significant increase in Na+/gamma aminobutyric acid cotransport. The activity of this system, which also transports betaine, remained unchanged after just five hours of hyperosmotic stress. Inclusion of betaine in the hyperosmotic medium reduced up-regulation of system A. Na/Pi cotransport also was up-regulated by five hours of hyperosmotic stress. Up-regulation of system A, but not Na/Pi cotransport, was detected in isolated membrane fractions indicating that increased activity of this membrane transport system may be one mechanism by which vascular endothelial cells accumulate amino acids. The amino acids may act as organic osmolytes to help maintain normal cell volume during the early phase of hyperosmotic stress. PMID- 9350657 TI - Intracellular transport, cell-surface exposure and release of recombinant Tamm Horsfall glycoprotein. AB - Human Tamm-Horsfall glycoprotein (T-H), first described as the major urinary glycoprotein, is a glycosylphosphatidyl-inositol (GPI)-anchored membrane protein which mainly resides at the luminal face of cells of the thick ascending limb of Henle's loop (TAL) and early distal convoluted tubules of nephron. Since no human renal cell-line producing T-H is available, T-H cDNA was transfected in HeLa cells and a cell line was selected in which 95% of the cells stably expressed T H, in order to elucidate the biosynthesis, mechanisms regulating the transport of T-H along the exocytic pathway, exposure at the cell surface and release in soluble form. Treatment of cells with an exogenous reducing agent results in a drastic delay in the conversion from precursor to mature T-H. Since the accumulating T-H-precursor carries glycans not yet processed by Golgi mannosidases, we propose that the formation of a correct set of intrachain disulphide bonds is required for T-H exit out the endoplasmic reticulum. Even the treatment of cells with an inhibitor of GPI-anchor biosynthesis results in an intracellular accumulation of T-H precursor, loss of T-H localization into Golgi apparatus and reduced surface exposure. These results indicate that the GPI anchor addition is necessary for T-H delivery to the cell-surface. The release rate of new synthesized T-H shows an initial lag time very likely depending on the time required for T-H surface exposure. A portion of released T-H appears to contain ethanolamine, a component of GPI anchor, indicating that, at least in HeLa cells, a GPI-specific phospholipase contributes to the T-H release. Exposure of cells to monensin and brefeldin A results in a loss of accumulation of T-H in the Golgi perinuclear region and a reduced delivery to the cell surface. Under monensin treatment an intermediate T-H form non-exposed at the cell surface is released in the medium, indicating that a soluble T-H may be produced inside the cell under conditions that alter the Golgi apparatus. If such an event occurs in polarized kidney cells, a T-H release from the basolateral face may be postulated, inasmuch as the GPI-anchor is an apical sorting signal. Since T-H is a powerful autoantigen, the accumulation of soluble T-H in the interstitium of TAL may cause the formation of immunocomplexes. PMID- 9350658 TI - Indomethacin and staurosporine reverse alpha 2 inhibition of water transport in rat IMCD. AB - These studies were conducted to determine if the prostaglandin-synthesis inhibitor indomethacin or the protein kinase C (PKC) inhibitor staurosporine affect the inhibition of osmotic water permeability (Pf) by the alpha-2 (alpha 2) agonist dexmedetomidine in the rat inner medullary collecting duct (IMCD). Terminal IMCDs from Wistar rats were perfused and Pf was increased with either 220 pM arginine vasopressin (AVP) or 0.1 mM 8-chlorophenylthio cyclic adenosine monophosphate (8CPTcAMP). All agents were added to the bathing solution. Dexmedetomidine at 100 nM inhibited both AVP- and 8CPTcAMP-stimulated Pf. When Pf was increased by AVP, indomethacin at 0.1 mM or 5 microM reversed the dexmedetomidine-induced inhibition by 68% and 43%, respectively. When Pf was increased by 8CPTcAMP, indomethacin at 0.1 mM or 5 microM reversed inhibition by 83% and 70%, respectively. Indomethacin increased AVP and 8CPTcAMP-stimulated Pf by 20 to 30% and dexmedetomidine inhibited the AVP+ indomethacin-stimulated Pf. Staurosporine at 10 nM yielded similar results. Results suggest that PKC and prostaglandins are involved in the alpha 2 mediated mechanism, and staurosporine and indomethacin-sensitive cellular mediators modulate basal Pf. PMID- 9350659 TI - Measurement of renal vein blood flow in rats by high-field magnetic resonance. AB - The aim of the present study was to examine whether magnetic resonance imaging (MRI) based method for non-invasive in vivo measurement of vein blood flow in rats could be used to estimate renal blood flow (RBF). Measurements were performed using a high-field (7 Tesla) MRI scanner with a short echo time phase contrast velocity measurement pulse sequence. The method was evaluated in vitro by flow measurements in an acrylic pipe and in vivo by recording left renal vein blood flow in normal and unilaterally nephrectomized rats. In a subset of animals RBF was measured by a direct method using 14C-tetraethylammoniumbromide. In vitro a high accuracy was found between applied and MRI measured flow rates in the range from 0.5 to 33 ml/min (r = 0.997; P < 0.001). In vivo the MRI measured left renal vein blood flow was 70% higher in unilaterally nephrectomized animals compared to control animals (3.4 +/- 0.4 ml/min/ 100 g body wt vs. 2.0 +/- 0.1 ml/min/100 g body wt, P < 0.001). Direct measurements of RBF revealed comparable values (3.4 +/- 0.3 ml/min/100 g body wt vs. 2.3 +/- 0.4 ml/min/100 g body wt, P = 0.05). In addition, the left kidney volume was recorded by MRI with an increase amounting to 40% (1.18 +/- 0.05 ml vs. 0.84 +/- 0.02 ml; P < 0.001) in the nephrectomized group compared to controls. Finally, a positive correlation was seen between left renal vein blood flow and MRI measured renal volume (r = 0.91; P < 0.001). In summary, MRI is a non-invasive tool by which measurement of renal vein blood flow can be performed, and it is concluded that MRI-based renal vein flow measurements can be used to estimate RBF in small rodents. PMID- 9350660 TI - Pressure diuresis and natriuresis in DOCA-salt mice. AB - The increasing use of mice in renal and cardiovascular studies has necessitated adapting physiological methods used for rats to mice, which are far smaller in size. We have adapted measurements of continuous renal blood flow, pressure natriuresis and diuresis, and laser-Doppler cortical and medullary flow to 40 g mice with DOCA-salt hypertension. We demonstrated a rightward shift in the pressure-natriuresis-diuresis curve. We conclude that with current, commercially available equipment, sophisticated renal physiology can be conducted in the mouse. These methods will be important to investigations of gene-targeted mice. PMID- 9350661 TI - Impaired autoregulation of GFR in hypertensive non-insulin dependent diabetic patients. AB - We investigated the effect of acute lowering of blood pressure (BP) upon glomerular filtration rate (GFR) in hypertensive non-insulin-dependent diabetes mellitus (NIDDM) patients, 14 with diabetic nephropathy and 12 with normoalbuminuria. The study was performed twice with the subjects receiving an intravenous injection of either clonidine (150 to 225 micrograms) or saline (0.154 mmol/liter). We assessed GFR, albuminuria, and BP. The two groups were well matched with respect to demographic data, baseline GFR and BP. Clonidine induced similar reductions in mean arterial blood pressure 19 (SE +/- 4) and 21 (SE +/- 3) mm Hg in patients with and without nephropathy, respectively. In the nephropathy group GFR diminished in average from 90 (SE +/- 6) to 81 (SE +/- 7) ml/min/1.73 m2 (P = 0.006), fractional clearance of albumin (x 10(-6)) declined from a geometric mean of 219 (antilog SE /divided by 1.3) to 186 (antilog SE /divided by 1.3) (P = 0.04), and four patients had a complete pressure-passive vasculature, defined as delta GFR% = delta MABP%. A significant correlation between relative reductions in MABP and GFR (r = 0.78, P < 0.001) was demonstrated in albuminuric patients. None of the normoalbuminuric patients had a complete pressure-passive vasculature and there were no significant differences in GFR between the two examinations, but five had abnormal autoregulation of GFR. Mean difference between changes in GFR (95% confidence interval) between the nephropathic and normoalbuminuric group was 5.5 (divided by 2.7 to 13.7) ml/min/1.73 m2 (P = 0.18). Our study suggests that hypertensive NIDDM patients, particularly patients with nephropathy, frequently suffer from impaired or abolished autoregulation of GFR. PMID- 9350662 TI - Antibodies to C1q in systemic lupus erythematosus: characteristics and relation to Fc gamma RIIA alleles. AB - Autoantibodies to the collagen-like region of the first complement component (C1qAB) are found in patients with systemic lupus erythematosus (SLE), particularly those with renal disease. In a cohort of 46 SLE patients with diffuse proliferative glomerulonephritis, we found declining C1qAB titers in 77% of treatment responders and in only 38% of treatment non-responders (P < 0.03). To further characterize this autoantibody, we tested 240 SLE patients for the presence of C1qAB. Positive titers were found in 44% of patients with renal disease and 18% of patients without renal disease (chi2 P < 0.0003). Analysis of IgG subclass revealed IgG2 C1qAB alone in 34%, IgG1 C1qAB alone in 20%, and both IgG1 and IgG2 in 46% of patients. Fewer than 10% of patients had measurable titers of IgG3 or IgG4 C1qAB. The pathogenic role of these IgG2-skewed C1qAB may relate to impaired immune complex clearance by the mononuclear phagocyte system: IgG2 antibodies are efficiently recognized by only one IgG receptor, the H131 allele of Fc gamma RIIa (Fc gamma RIIa-H131). In contrast, Fc gamma RIIa-R131, which is characterized by minimal IgG2 binding, has recently been associated with lupus nephritis. In our C1qAB positive patients, the presence of Fc gamma RIIA R131 was associated with an increased risk for renal disease. Autoantibodies to C1q may have pathogenic significance in SLE patients with genetic defects in the ability to clear IgG2 containing immune complexes. PMID- 9350663 TI - Calcium channel blockers correct in vitro mitochondrial toxicity of cellulose acetate. AB - We studied the action of rinse solutions from cellulose acetate hemodialyzers on isolated mitochondria. We showed that concentrates from the rinses impaired the adenosine 5'-triphosphate (ATP) synthesis as reflected by the decrease in respiration during state 3 and in P/O ratio. This impairment results from a calcium release from mitochondria that is induced by rinse solution concentrates. The release, triggering the mitochondrial calcium carrier, would explain the decrease in ATP synthesis. Moreover, rinse solution concentrates hinder mitochondrial calcium storage. The rise in cytosolic calcium in hemodialyzed patients may be related, at least in part, to these findings, since a lack of ATP impairs the ATP-dependent cellular calcium-extrusion pumps. We also showed that calcium channel blockers, at therapeutically relevant doses, restore ATP synthesis and calcium storage in mitochondria impaired by rinse solution concentrates. Finally, these in vitro results were confirmed by experiments on cells in culture proving that Diltiazem counteracts the cytotoxicity of rinse solution concentrates. These findings are consistent with observations that these drugs suppress the increase in leukocyte cytosolic calcium in dialyzed patients. Moreover, this would help explain the efficiency of calcium channel blockers in cells without L-calcium channels. PMID- 9350664 TI - Recombinant human erythropoietin enhances superoxide production by FMLP stimulated polymorphonuclear leukocytes in hemodialysis patients. AB - Recombinant human erythropoietin (rHuEPO) is a hematopoietic growth factor that has a broad spectrum of action. We have observed the in vivo and in vitro effects of rHuEPO on the superoxide production of circulating polymorphonuclear leukocytes (PMNs) in hemodialysis patients. The PMNs were separated from heparinized blood after dextran sedimentation and Ficoll-Conray centrifugation and stimulated with formyl-methionyl-leucyl-phenylalanine (FMLP), serum-treated zymosan (STZ), or phorbol myristate acetate (PMA). The in vivo study showed that rHuEPO therapy for 12 weeks enhanced the superoxide production by FMLP-stimulated PMNs (P < 0.01). However, no significant changes on superoxide production was found in either STZ- or PMA-stimulated PMNs. Simultaneous measurement of PGE2 production by PMNs in response to all three stimulants did not show any significant changes after rHuEPO therapy. The direct in vitro effect of rHuEPO on PMNs showed that rHuEPO does not enhance the superoxide production by non stimulated PMNs. However, preincubation of rHuEPO enhanced superoxide production from FMLP- and STZ-stimulated PMNs. Our results indicate that rHuEPO enhanced FMLP-stimulated superoxide production of PMNs both in vivo and in vitro in hemodialysis patients, which may be responsible for the increased oxidant stress in hemodialysis patients after rHuEPO therapy. PMID- 9350665 TI - Comparison of methods to predict equilibrated Kt/V in the HEMO Pilot Study. AB - The ongoing HEMO Study, a National Institutes of Health (NIH) sponsored multicenter trial to test the effects of dialysis dosage and membrane flux on morbidity and mortality, was preceded by a Pilot Study (called the MMHD Pilot Study) designed to test the reliability of methods for quantifying hemodialysis. Dialysis dose was defined by the fractional urea clearance per dialysis determined by the predialysis BUN and the equilibrated postdialysis BUN after urea rebound is completed (eKt/V). In the Pilot Study the blood side standard for eKt/V was calculated from the predialysis, postdialysis, and 30-minute postdialysis BUN. Four techniques of approximating eKt/V that eliminated the requirement for the 30-minute postdialysis sample were also evaluated. The first adjusted the single compartment Kt/V using a linear equation with slope based on the relative rate of solute removal (K/V) to predict eKt/V (rate method). The second and third techniques used equations or mathematical curve fitting algorithms to fit data that included one or more samples drawn during dialysis (intradialysis methods). The fourth technique (dialysate-side) predicted eKt/V from an analysis of the time-dependent profile of dialysate urea nitrogen concentrations (BioStat method; Baxter Healthcare, Inc., Round Lake, IL, USA). The Pilot Study demonstrated the feasibility of conventional and high dose targets of about 1.0 and 1.4 for eKt/V. Based on the blood side standard method, the mean +/- SD eKt/V for patients randomized to these targets was 1.14 +/- 0.11 and 1.52 +/- 0.15 (N = 19 and 16 patients, respectively). Single-pool Kt/Vs were about 0.2 Kt/V units higher. Results were similar when eKt/V was based on dialysate side measurements: 1.10 +/- 0.11 and 1.50 +/- 0.11. The approximations of eKt/V by the three blood side methods that eliminated the delayed 30-minute post-dialysis sample correlated well with eKt/V from the standard blood side method: r = 0.78 and 0.76 for the single-sample (Smye) and multiple-sample intradialysis methods (N = 295 and 229 sessions, respectively) and 0.85 for the rate method (N = 295). The median absolute difference between eKt/V computed using the standard blood side method and eKt/V from the four other methods ranged from 0.064 to 0.097, with the smallest difference (and hence best accuracy) for the rate method. The results suggest that, in a dialysis patient population selected for ability to achieve an equilibrated Kt/V of about 1.45 in less than a 4.5 hour period, use of the pre and postdialysis samples and a kinetically derived rate equation gives reasonably good prediction of equilibrated Kt/V. Addition of one or more intradialytic samples does not appear to increase accuracy of predicting the equilibrated Kt/V in the majority of patients. A method based on dialysate urea analysis and curve-fitting yields results for equilibrated Kt/V that are similar to those obtained using exclusively blood based techniques of kinetic modeling. PMID- 9350666 TI - Percutaneous recanalization of occlusion of central and proximal veins in chronic hemodialysis. Technical note. AB - Occlusion of the central and proximal veins in chronic hemodialysis patients results in considerable edema of the arm of the vascular access that is unable to drain normally. This is a formidable problem because it is very often necessary to close the vascular access, which is sometimes the last available one. To avoid resorting to this disastrous solution, recanalization of the occluded vein by percutaneous recanalization followed by endoluminal angioplasty was successfully performed in five patients (4 innominate veins and one axillary vein). Immediate failure occurred in a sixth patient, and delayed failure after two months of patency (innominate vein) in another patient for whom there had been no systematic stent placement. Recanalization was still patent in four other patients at 3, 6, 12 and 26 months. These results are an encouragement to attempt percutaneous recanalization by angioplasty of occluded central veins because, when successful, this technique makes it possible to preserve the vascular access and to avoid onerous surgery. We believe that this technique should therefore become better known. PMID- 9350667 TI - Disorders of bone and mineral metabolism after renal transplantation. PMID- 9350668 TI - An update on uremic toxins. AB - The author reviews trends and evolution in biochemical methodology, clinical symptomatology description, experimental models and definitions of uremic toxins. The assumption is made of one specific toxic effect for one specific toxin. PMID- 9350669 TI - Parathyroid hormone, chronic renal failure and the liver. PMID- 9350670 TI - p-cresol and uric acid: two old uremic toxins revisited. PMID- 9350671 TI - Uremic cardiomyopathy: role of parathyroid hormone. PMID- 9350672 TI - Indoxyl sulfate increases the gene expressions of TGF-beta 1, TIMP-1 and pro alpha 1(I) collagen in uremic rat kidneys. AB - We recently reported that the serum levels of indoxyl sulfate, a dietary protein metabolite, are increased in both uremic rats and patients, and that the administration of indoxyl sulfate to uremic rats accelerates the progression of glomerular sclerosis. Thus, we hypothesize that the overload of protein metabolites such as indoxyl sulfate on nephrons promotes the progression of chronic renal failure (CRF). Recent studies revealed that tubulointerstitial injury is of equal or greater importance than glomerular sclerosis in determining whether progressive renal dysfunction will ensue in various renal diseases. In the present study, to clarify the role of indoxyl sulfate in the progression of CRF, the expressions of genes related to tubulointerstitial fibrosis such as transforming growth factor (TGF)-beta 1, tissue inhibitor of metalloproteinases (TIMP-1) and pro-alpha 1(I) collagen were examined in the renal cortex of 5/6 nephrectomized uremic rats given indoxyl sulfate. In the first experiment, the administration of indoxyl sulfate for five weeks significantly increased the mRNA levels of TGF-beta 1, TIMP-1 and pro-alpha 1(I) collagen in the uremic rats given indoxyl sulfate compared with the control uremic rats, accompanied by a significant decline in renal function and worsening of glomerular sclerosis. In the second experiment, the administration of indoxyl sulfate for 2.5 weeks also increased the expression of the mRNA levels with no significant decline in the renal function. In conclusion, these findings indicate that the overload of the protein metabolite indoxyl sulfate on remnant nephrons is involved in the increased bioactivity of TGF-beta 1 in uremic kidneys, which enhances the renal expression of TIMP-1 and type 1 collagen, leading to the progression of CRF. PMID- 9350673 TI - The protein metabolite hypothesis, a model for the progression of renal failure: an oral adsorbent lowers indoxyl sulfate levels in undialyzed uremic patients. AB - We have recently demonstrated that indoxyl sulfate promotes the progression of glomerular sclerosis in uremic rats. In the present study, we determined whether an oral adsorbent (AST-120) could reduce the serum and urine levels of indoxyl sulfate and suppress the progression of chronic renal failure (CRF) in undialyzed uremic patients. Twenty-five undialyzed uremic patients were given AST-120 at a dose of 6 g/day for 6 months, while 10 undialyzed uremic patients were not given AST-120. The effects of the oral adsorbent on the slope of the 1/serum creatinine (Scr)-time plot, and the serum and urine levels of indoxyl sulfate were evaluated. Administration of AST-120 significantly decreased the serum and urine levels of indoxyl sulfate, and tended to improve the slope of the 1/SCr-time plot in the CRF patients. Among the patients in whom urinary excretion of indoxyl sulfate was reduced by AST-120, the oral adsorbent significantly improved the slope of the 1/SCr-time plot. The change in the slope of the 1/SCr-time plot showed a significant negative correlation with the change in the urine level of indoxyl sulfate. Thus, patients who showed a greater decrease of urinary indoxyl sulfate also showed more marked suppression of the progression of CRF. These results support the notion that indoxyl sulfate, a protein metabolite, is involved in the progression of CRF, and that an oral adsorbent can delay progression at least partly by reducing the serum and urine levels of indoxyl sulfate. PMID- 9350674 TI - Role of increased glomerular protein traffic in the progression of renal failure. AB - Clinical and experimental data have indicated that heavy proteinuria in renal glomerular diseases is associated with the formation of tubulointerstitial fibrosis and contributes to the progression of renal failure. In recent years studies have focused on the possibility that albumin and other proteins that accumulate in the lumen of proximal tubular cells as a consequence of glomerular permeability dysfunction, are a direct cause of tubular cell injury. Specific proteins that have been shown to be cytotoxic are transferrin/iron, lipoproteins and complement components, all of which appear in the urine in proteinuric states. As an additional pathway of injury one may consider the effects of lipids bound to albumin and lipoproteins, including oxidized low density lipoproteins, which, by inducing an oxidative stress to tubular cells, are potent cytotoxic molecules. Moreover, reabsorption of high molecular weight proteins activates proximal tubular cells to produce matrix proteins, cytokines, chemoattractants and vasoactive mediators that may-converge in stimulating interstitial inflammation and scarring. Given the functional toxicity of filtered proteins on the kidney, pharmacological and dietary manipulations aimed at reducing glomerular protein traffic may have a beneficial impact on the deterioration of renal function in progressive nephropathies. PMID- 9350675 TI - Renal fibrosis: role of impaired proteolysis and potential therapeutic strategies. PMID- 9350676 TI - Lipids in progression of renal disease. AB - There is increasing evidence for the role of hyperlipidemia as a contributing factor to the progression of chronic renal disease. Experimental studies in animal models suggest that lipids might be important modulators in progressive renal disease. On one hand maneuvers designed to elevate cholesterol worsen renal disease while reduction of lipid-lowering agents have demonstrated beneficial effects. In humans, correcting abnormalities in lipid metabolism with antilipemic therapy may help slow the rate of functional decline in patients with progressive renal disease as well as reduce the frequency of acute rejection after renal transplantation. However, large controlled clinical trials examining the effect of lipid-lowering strategies on progression of renal disease have not been carried out. Further clinical trials delineating the precise role of antilipemic agents in treating patients with renal disease appears warranted. PMID- 9350677 TI - Mechanism of the progression of diabetic nephropathy to renal failure. AB - Although the evolution of diabetic nephropathy is brought about mostly by persistent hyperglycemia, its progression may be influenced by various other factors such as hypertension and dietary protein intake. It has been recently suggested in the literature that the gene polymorphism of angiotensin converting enzyme (ACE) might be associated with the development of diabetic nephropathy, because the DD genotype of ACE gene is closely associated with the presence of nephropathy in diabetic subjects. However, in our present analysis the frequency of the DD genotype in patients with non-insulin dependent diabetes is not significantly related to the presence or absence of nephropathy. It remains to be clarified by multi-center analysis using large numbers of patients whether the gene polymorphism of ACE is related to the progression of diabetic nephropathy to renal failure. Furthermore, it has been postulated that the interstitial fibrosis evaluated in renal biopsy specimens is significantly correlated with the declining of renal function in diabetic patients. However, it is not possible to clinically quantitate the interstitial fibrosis without performing renal biopsy. We have recently found that the urinary excretion of type IV collagen is significantly increased in diabetic patients. Moreover, the increase in urinary type IV collagen is well correlated with the amount of urinary albumin. Since type IV collagen in the urine is probably derived from tubulointerstitial tissue, it is likely that the increased amount of type IV collagen in the urine may reflect the fibrotic change in diabetic kidneys. Whether the increase in urinary type IV collagen is able to predict for the progression of diabetic nephropathy in the future should be examined. PMID- 9350678 TI - Mechanisms of malnutrition in uremia. AB - The pathogenesis of protein wasting in chronic renal failure is multifactorial. Potential mediators of protein catabolism in chronic uremia include anorexia, low protein-energy intake, increased cortisol and parathyroid hormone secretion, insulin resistance, metabolic acidosis and unidentified uremic toxins. In non acidotic uremic patients the rate of protein turnover (that is, synthesis and degradation) has often been found to be decreased. Malnutrition also decreases both protein synthesis and degradation. In contrast, during acidosis protein degradation is primarily accelerated and results in rapid loss of body proteins. Cytokine concentrations have often been found increased in both dialyzed and undialyzed chronically uremic patients. Our study determined the circulating levels of TNF-alpha and of type I (60 kDa) and type II (80 kDa) soluble TNF-alpha receptors in undialyzed uremic patients, and found that their plasma levels were greatly increased. Serum creatinine correlated with TNF-alpha soluble receptors but not with the TNF-alpha. Thus, TNF-alpha is potentially an important mediator of protein wasting in chronically uremic patients. Pharmacological therapy of protein catabolism in chronic uremia may include the administration of pentoxifylline, which has been shown to decrease protein degradation by interfering with the TNF-alpha system (that is, TNF-alpha and its soluble receptors) in experimental models. Growth hormone and insulin-like growth factor 1 administration may also be beneficial in these patients, but further evaluation of the hormone effects on glucose and glutamine metabolism is called for. PMID- 9350680 TI - Carbohydrate and insulin metabolism in renal failure. PMID- 9350679 TI - IGF-1 resistance in chronic renal failure: current evidence and possible mechanisms. PMID- 9350682 TI - Metabolism of vitamin B6 and its requirement in chronic renal failure. AB - Vitamin B6 is very important for the normal function of multiple organ systems. In the majority of patients with chronic renal failure and in patients during various forms of renal replacement therapy can develop vitamin B6 deficiency from many causes. Intravenous administration of 20 mg furosemide led to the increase of urinary excretion and fraction excretion (FE) of vitamin B6 in patients with chronic renal failure. This is a new side effect of furosemide. The daily oral dose of pyridoxine 6 mg was optimal for the patients without erythropoietin (EPO) treatment during the period of 12 months of CAPD. Erythrocyte vitamin B6 was determined by an indirect method, that is, by measuring the effect of pyridoxal-5 phosphate (PLP). In the other group of CAPD patients plasma vitamin B6 was in the reference range, and the mean value of peritoneal clearance of vitamin B6 was very low: 8.8% of urea clearance. In addition, an indirect relationship between the effect of PLP and plasma vitamin B6 was found. Indirect evidence has shown that erythrocyte vitamin B6 is consumed by the hemoglobin synthesis much more during EPO treatment in hemodialysis patients. No influence of pyridoxine 5 to 6 mg/day on decreased parameters of cellular immunity was found. For prevention of vitamin B6 deficiency in hemodialysis and CAPD patients we recommend the following doses of pyridoxine: for patients without EPO treatment 5 mg/day, and with EPO treatment 20 mg/day. A favorable effect of pyridoxine 50 mg/day has also been found on several parameters of cellular immunity in hemodialysis patients. PMID- 9350681 TI - Inflammation, dyslipidemia and vascular risk factors in hemodialysis patients. AB - Cardiovascular complications account for more than 50% of death in hemodialysis patients. Strong and independent predictors of mortality or cardiovascular complications are low levels of serum albumin, high plasma C-reactive protein and lipoprotein(a), plasma proteins that are described to function as negative or positive acute phase reactants. Further prominent and known risk factors that contribute to the increased incidence of atherosclerosis in hemodialysis patients are disorders in lipoprotein metabolism and elevated plasma fibrinogen concentrations. The latter has also been described to increase following acute or chronic inflammation. The main metabolic abnormality of the lipoprotein profile is a delayed catabolism of triglyceride-rich apoB-containing lipoproteins caused by a decreased activity of lipolytic enzymes. Inhibition of lipoprotein lipase activity by cytokines or parathyroid hormone impedes conversion of very-low density lipoprotein to low-density lipoprotein, resulting in remnant accumulation and hypertriglyceridemia. Another acute phase condition, namely, acute myocardial infarction, results in a similar pattern of dyslipidemia and coagulation disorder. In summary, the acute phase response deeply influences serum lipids and lipoproteins as well as other atherogenic acute phase proteins in hemodialysis patients. Appreciation of acute phase lipoprotein changes is essential for accurate diagnosis of dyslipidemias, proper design of future clinical studies, and correct interpretation of published data. PMID- 9350683 TI - Resistance of the parathyroid glands to vitamin D in renal failure: implications for medical management. PMID- 9350684 TI - Uremic toxins and vitamin D metabolism. PMID- 9350685 TI - Is the bone mass of hemodialysis patients genetically determined? AB - Polymorphism of the vitamin D receptor gene (VDR) has been linked to bone mineral density in twins, postmenopausal osteoporosis, and premenopausal woman. We examined the possibility that the bone mass in hemodialysis (HD) patients might be determined by VDR. The study consisted of 229 HD patients with a mean age of 53.3 years (range 21 to 83), who were dialyzed three times a week for an average of 8.65 (range 0.2 to 24) years. We determined their VDR using DNA of peripheral white blood cells by restriction enzyme BsmI and the polymerase chain reaction restriction fragment length polymorphism method. Bone mineral content (BMC) was estimated at 1/3 of the radius using dual energy X-ray bone absorptiometry, and expressed in z-scores standardized by gender and age. Distributions of VDR in this hemodialysis population were BB (9.9%), Bb (13.1%), and bb (77.0%), showing no significant different from those in 105 healthy volunteers (BB, 7.6%; Bb, 13.3%; and bb, 79.0%). Multiple regression analysis revealed that gender, age, duration on HD, and serum osteocalcin are major determinants of BMC (r = 0.762, P < 0.001), while VDR and serum parathyroid hormone are not. In a subgroup with younger (< 65 years) patients dialyzed for less than 8.65 years, the z-score of BMC of patients with BB allele was less than those with Bb and bb allele (N = 77, P = 0.020). We conclude that vitamin D receptor polymorphism is not one of the main determinants of BMC of HD patients, though it might partially effect bone mass in a subgroup of younger HD patients with shorter HD histories. Further studies with longitudinal observation will be needed to confirm these possibilities. PMID- 9350686 TI - Incidence and clinical characteristics of hypoparathyroidism in dialysis patients. AB - The incidence and clinical characteristics of hypoparathyroidism (Hypo) were evaluated in 8188 hemodialysis (HD) and 1207 CAPD patients treated in 65 hospitals or clinics in Japan. Hypo was defined by an intact parathyroid hormone (PTH) level below 160 pg/ml, which corresponded to the low normal limit of intact PTH to maintain a normal osteoblastic surface in 40 bone biopsy specimens of Japanese dialysis patients, and patients were classified into two groups: absolute Hypo (A-Hypo), intact PTH < 60 pg/ml, and relative Hypo (R-Hypo), 60 pg/ml < or = intact PTH < 160 pg/ml. A total of 2537 (31.0%) and 2736 (33.4%) HD patients were classified into A- and R- Hypo, and 401 (31.3%) and 379 (31.4%) CAPD patients occupied A- and R-Hypo, respectively. A high incidence of Hypo was observed in the HD patients with diabetes mellitus (DM) and old age (> or = 70 years old) compared with that of a nationwide epidemiological report for dialysis patients by Japanese Society for Dialysis Therapy. Hypo patients who were treated by CAPD had a background of being younger, having a shorter duration on dialysis, and were less frequently diagnosed with DM than in those Hypo patients on HD. Bone pain and metastatic calcification were observed in approximately 20% and 25% of Hypo patients, respectively. No difference was observed in the background factors and the prevalence of signs and symptoms between the A- and R-Hypo groups, regardless of the mode of treatment (HD or CAPD). These results suggest that a very high incidence and specific backgrounds (DM and aging) of Hypo exist in Japanese dialysis patients. PMID- 9350689 TI - Evidence that the anemia of renal failure participates in overall uremic toxicity. PMID- 9350688 TI - Immune dysfunction in uremia. AB - Among uremic patients on hemodialysis and continuous ambulatory peritoneal dialysis treatment infectious complications leading to a high incidence of morbidity and mortality are a well documented problem. Polymorphonuclear leukocytes (PMNLs) are the main cells of the unspecific defence system during bacterial infections. There is a number of partly interdependent factors responsible for the diminished PMNL functions (chemotaxis, phagocytosis, intracellular killing by proteolytic enzymes and toxic oxygen radicals) found in uremia: iron overload, elevated levels of intracellular calcium and hemodialysis treatment per se has been shown to be involved in altered PMNL functions. Uremic toxins are circulating plasma factors accumulating in the serum of uremic patients. They are thought to play a crucial role in inhibiting the unspecific immune defence. A number of uremic toxins has already been purified and characterized. In our laboratory, a granulocyte inhibiting protein (GIP) with homology to immunoglobulin light chains has been isolated. We could show that free immunoglobulin light chains per se are able to interfere with essential PMNL functions. A GIP with homology to beta 2-microglobulin was also isolated from dialysis patients. Angiogenin was purified from uremic patients as a PMNL degranulation inhibiting protein and complement factor D was shown to adversely affect PMNL functions. A modified form of ubiquitin isolated from peritoneal dialysis patients interferes with PMNL chemotaxis. Furthermore, p-cresol was identified as a uremic solute that impairs the respiratory burst activity of PMNL. There is also increased clinical evidence for profound defects in the specific immune defence in uremia, such as the high susceptibility to viral infections in uremic patients, the deficient responses of their T lymphocytes, and the significantly depressed specific antibody responses. PMID- 9350687 TI - Apolipoprotein E and alpha 1-antichymotrypsin in dialysis-related amyloidosis. AB - Dialysis-related amyloidosis as represented by carpal tunnel syndrome is a serious complication of long-term dialysis treatment of patients with chronic renal failure. beta 2-microglobulin has been identified as a structural component of the amyloid deposits, but other factors also are associated with amyloid formation. We recently demonstrated the presence of apolipoprotein E and alpha 1 antichymotrypsin in the amyloid deposits. We therefore analyzed how polymorphic variants of both genes were related to the onset of amyloidosis. Among the apolipoprotein E genotypes, allele epsilon 2 represented a protective factor that delayed the onset of disease. In contrast, polymorphic alpha 1-antichymotrypsin alleles had no effect on the onset of amyloidosis. Thus, the apolipoprotein E epsilon 2 allele can be added to the list of factors that determine the onset of dialysis-related amyloidosis, which include patient age at initiation of dialysis therapy, dialysis duration, and the dialysis membrane used. PMID- 9350690 TI - Hemostasis disorders in chronic renal failure. AB - Dialysis patients with CRF show a fibrinolysis defect at the level of plasminogen activation. The fibrinolysis defect in CRF deepens as renal function declines. Reduced fibrinolysis may be responsible, along with other factors, for the development of thrombosis, atherosclerosis and their thrombotic complications. A number of important and relevant questions, which are more or less interrelated, continue to be left without clear-cut answers. These include, for instance, what is the relationship between fibrinolysis defects in CRF and "prothrombotic" metabolic disorders? To what extent, if at all, does hypofibrinolysis normalize during treatment of these metabolic disorders? Indeed, is there a causative relationship between the decrease in fibrinolysis and thrombotic complications in patients with CRF? PMID- 9350691 TI - Atherosclerosis in uremia: possible roles of hyperparathyroidism and intermediate density lipoprotein accumulation. AB - Cardiovascular motality is high in patients with chronic renal failure treated with dialysis, and secondary hyperparathyroidism may promote atherosclerogenesis. Recent studies have revealed advanced atherosclerosis in hemodialysis patients by using high-resolution B-mode ultrasonography. Multiple regression analyses indicated that hyperphosphatemia and hyperparathyroidism were associated with increased intima-media thickness (IMT) of the carotid and femoral arteries in hemodialysis patients, respectively. Hypocalcemia and hyperparathyroidism independently and adversely affect the lipoprotein profile by suppressing hepatic triglyceride lipase (HTGL), a lipid-regulating enzyme playing important roles in the metabolism of intermediate density lipoprotein (IDL) and high density lipoprotein (HDL). Plasma IDL is raised markedly, and HDL is lowered in uremia. These lipoprotein changes are closely associated with increased aortic pulse wave velocity (PWV), an index of aortic sclerosis. These findings support the hypothesis that deranged calcium-phosphate homeostasis and secondary hyperparathyroidism promote atherosclerosis in uremia, at least partly by affecting lipoprotein metabolism. Adequate dialysis and efforts to normalize calcium, phosphate and PTH would be beneficial in preventing not only bone disease, but atherosclerosis as well. PMID- 9350692 TI - Current trends in dialysis therapy. PMID- 9350693 TI - Adequacy of dialysis. AB - Despite improvements in the delivery of dialysis, morbidity and mortality remain excessively high in end-stage renal disease patients. Multiple lines of evidence suggest that inadequate dialysis is responsible these trends. The limits of our understanding of the influence of dialysis dose on mortality and morbidity are discussed. In that context, the relationship between dialysis dose and nutritional status of dialysis patients is also reviewed. The best marker for adequacy of dialysis is yet to be determined. However, urea as a surrogate for small molecular weight solute clearance, has been demonstrated by many studies to be a parameter of adequacy, both in hemodialysis and peritoneal dialysis populations. The application of kinetic modeling of urea is discussed for hemodialysis. The new insights that will be forthcoming at the conclusion of the National Institute of Health HEMO study should help to define an optimal dose of dialysis. PMID- 9350694 TI - Are biocompatible membranes superior for hemodialysis therapy? PMID- 9350695 TI - Issues affecting the longevity of the continuous peritoneal dialysis therapy. AB - Continuous ambulatory peritoneal dialysis (CAPD) is one of the accepted dialysis modalities worldwide. However, the longevity of the CAPD technique and the patient survival associated with this modality have not been established. We review our 16 year experience in one treatment center in 224 patients who underwent CAPD. Overall, these patients survived on CAPD for a mean of 6.6 years; a 50% survival was reached at a mean of 5.5 years. Ultrafiltration loss was the most prominent cause of withdrawal from CAPD among patients in whom the technique was successful for more than 6 years. Countermeasures to decrease the CAPD associated morbidity are crucial to prolonging the success of the technique, and thus increasing the duration of patient survival. PMID- 9350696 TI - New frontiers in continuous ambulatory peritoneal dialysis. PMID- 9350697 TI - Salt: a sacred substance. AB - Salt is the last relic of the ocean where life was born. Its presence has influenced the whole gamut of history and its name is linked to hundred of geographical locations. Its importance for nutrition is supported by the discovery of Aeneolithic salt cellars. Salt cellars and pyramids of salt have been included in paintings and other works of art. In Japan where salt was and still is obtained from the sea, a salt culture has developed that can be traced in the rituals of everyday life, including meal preparation, sports, and Shinto ceremonies. PMID- 9350698 TI - Report on primate supply for biomedical scientific work in the UK. EUPREN UK Working Party. AB - A Working Party of the UK group of European Primate Resources Network (EUPREN) considered primate supply for scientific work in the UK. Through a questionnaire, which achieved a very good response, it obtained details of primate use, sources and breeding in the UK and it put forward options to ensure that animal welfare is the best possible whilst ensuring continued supply. The questionnaire showed that contract research laboratories and pharmaceutical companies use about 80% of the 4233 primates used annually at the moment, with the rest accounted for by academic establishments and public sector laboratories. Fifty-four per cent are cynomolgus macaques (Macaca fascicularis), of which nearly 90% are captive-bred outside the European Union (EU), the remainder being bred in the UK. Nearly 90% of cynomolgus macaques are used by only five institutions. Thirty-seven per cent of primates used are marmosets (Callithrix jacchus jacchus), all of which are bred in the UK. Most of the rest are rhesus macaques (Macaca mulatta), about half of which are captive-bred outside the EU, the other half being bred in the UK. Overall primate use has increased from about 3000 per year in 1990 and users predict that requirements for all species except baboons (Papio sp.) will be maintained or increase. Marmoset breeding in the UK is already closely matched to use, and it could be increased reasonably easily if necessary. Some of the existing breeding centres of macaques in the UK would be prepared to consider expanding to supply others, although investment and imported breeding stock would be needed and it is likely that a large investment would be needed to breed a significant fraction of the macaque use in the UK. A further problem is that the users of only about 10% of the cynomolgus macaques said that they could replace this species by rhesus macaques, which are easier to breed in the UK. The questionnaire showed that much of the use of macaques would be transferred to other countries equally remote from the natural source countries of the animals, if constraints on primate use became more severe in the UK. Users felt that it is unlikely that much of the work could be transferred to the natural source countries themselves. A review of the literature revealed a paucity of information on the effects of transport on primate welfare. The importance of obtaining this information before making decisions about alternative means of supply is stressed. Current schemes for the accreditation of primate breeders were reviewed. A list of options is presented for discussion. Users vary so much in their requirements that it is unlikely that one means of supply will be applicable to all. Animal welfare will benefit and supply will be more certain if cooperation between those concerned (preferably through the UK group of EUPREN) is maintained. PMID- 9350699 TI - Sanitary aspects of handling non-human primates during transport. Report of the Federation of European Laboratory Animal Science Associations (FELASA) Working Group on Non-human Primate Health accepted by the FELASA Board of Management, April 1997. PMID- 9350700 TI - Brief review of scientific studies of the welfare implications of transporting primates. AB - The transportation of primates has become an important welfare issue and the outcome of the debate over its cost to the animal will have effects on the future of medical research using these species. There is a paucity of scientific studies on transport relating to primates and the need for gathering of further scientific evidence is highlighted. PMID- 9350701 TI - Salivary cortisol: a non-invasive measure of hypothalamo-pituitary-adrenocortical activity in the squirrel monkey, Saimiri sciureus. AB - Salivary cortisol is a non-invasive and easy-to-assess measure of the activity of the hypothalamo-pituitary-adrenocortical (HPA) system. Here we report that salivary cortisol determination can be used in squirrel monkeys (Saimiri sciureus) to monitor variations in HPA system activity induced by both housing and experimental conditions. Saliva cortisol assessment has several advantages over blood cortisol analysis such as stress-free frequent sampling, laboratory independence and lower costs. Therefore, this non-invasive measure can be the method of choice in primatological research projects and routine programmes related to the well-being of these laboratory animals. PMID- 9350702 TI - Reporting animal use in scientific papers. AB - This paper reports the results of an examination of the 'methods' sections of a range of experimental research papers in biomedical science, focusing on the descriptions of animal use and housing. Detailed descriptions in the methods should enable replication, and also enable readers to judge scientific quality. Relatively few papers sampled gave adequate descriptions of housing conditions and many failed to give details of physiologically relevant variables such as weight of animals. Thirty per cent of papers omitted the number of animals used, and the deaths of animals (whether as part of the protocol, or accidental death) were not always recorded. Adequate reporting of the conditions of animal maintenance and use are important, both in relation to the quality of the science produced, and also because of public concerns about the ethics of animal experiments. PMID- 9350703 TI - Analgesics in mice used in cancer research: reduction of discomfort? AB - During the last decades, an increase is apparent in the use of analgesics for laboratory animals in situations where this was previously considered unnecessary. Mice with advanced tumours often show clear signs of discomfort which may be a result of chronic pain or a result of general ill-being. The syngeneic murine tumour model most frequently used in our experiments was used to investigate whether this discomfort can be reduced with an analgesic. Twenty DBA/2 mice bearing SL2 lymphoma were given 0.5 mg/kg buprenorphine (Temgesic) in food gel twice daily, 20 tumour-bearing mice were given control food gel at the same times. Indicators of well-being were monitored daily. These included behavioural parameters such as exploration, grooming, and posture; food and water consumption and fur quality. All mice showed a clear increase of discomfort with time: explorative behaviours and grooming decreased, while sitting in hunched posture increased. Food and water consumption and fur quality also decreased. Major significant differences between the buprenorphine treated group and the control group were not apparent. In conclusion, we could not document a positive effect or buprenorphine on discomfort in mice as evaluated by our scoring system. It remains possible that pain itself was not the primary cause of the discomfort in mice bearing these tumours, or that the analgesic effect of buprenorphine was insufficient under these circumstances. PMID- 9350704 TI - Extrusion of earthworm coelomocytes: comparison of the cell populations recovered from the species Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum. AB - Coelomocytes were extruded from three earthworm species: Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum. Featuring a simple low-vacuum holding device, the proposed methodology allows the recovery of cells with minimum risk of contamination by faecal material. The viability of O. tyrtaeum coelomocytes was highly reproducible (average 93%), with an average yield of 0.92 x 10(6) viable cells per earthworm. Cell viability for L. terrestris and E. fetida averaged approximately 68% but the cell yields were higher (respectively 1.67 x 10(6) and 1.28 x 10(6)). Large inter-individual differences in cell yields were observed with L. terrestris. Flow cytometric analyses indicated species to species differences in cell populations. Coelomocytes from E. fetida were the smallest with approximately 57% of the total viable cells recovered being monitored between 2 and 10 microns. Large granulated cells (> or = 20 microns) were detected in fairly large proportions in L. terrestris and O. tyrtaeum (approximately 52 and approximately 96%, respectively) while they were less abundant in E. fetida (approximately 9%). Using the vital dye neutral red to assess functional integrity, average cellular uptakes were significantly higher for L. terrestris and O. tyrtaeum than for E. fetida (2.94, 2.66 and 0.64 micrograms/2 x 10(5) cells, respectively). In summary, the extrusion methodology herein described is applicable for the recovery of coelomocytes from a wide range of earthworm sizes and species. Moreover, this study strengthens the fact that extruded coelomocytes could be used for the evaluation of cell dysfunction and/or cell death following an in vitro and/or in vivo treatment. PMID- 9350705 TI - Behavioural and cardiovascular responses of rats to euthanasia using carbon dioxide gas. AB - Our results showed more rapid falls in pulse rate and blood pressure in rats euthanized in a chamber precharged with carbon dioxide (CO2), when compared with rats euthanized more slowly, but death still took over 5 min in the former group. There was no behavioural evidence of pain or distress in either group during euthanasia. Initial ataxia and dyspnoea was punctuated by a lag before death, thus separating euthanasia into three clearly defined phases. All visual signs of death preceded complete vascular collapse by about 1 min in both groups, so we recommend that gas flow be maintained for at least 1 min after apparent death. PMID- 9350706 TI - Hypoxia-induced pulmonary hypertension in an optimized environment for the guineapig. AB - Prolonged exposure to hypoxia elicits a variety of time-related morphologic and physiologic changes in the pulmonary vasculature of mammals, including humans. The study of hypoxia-induced changes in rodents generally requires a prolonged exposure to 9% oxygen for a minimum of 10 days in an airtight chamber, which has only been generally described in the literature as large (200-400 l), sealed acrylic chambers. To assist in the search for better therapies for diseases associated with chronic hypoxia using animal models, we have custom-built an airtight chamber for hypoxic exposure of rodents, and characterized the effect of chronic hypoxia on functional and morphologic changes in the pulmonary vasculature of the guineapig using this system. This chamber has been designed to alleviate any unnecessary stress related to food or water intake, cleanliness and excess illumination to the animals during the hypoxic-exposure period. Chronic exposure of the guineapig to hypoxia (0-21 days) produced time-related physiologic, morphologic, and haematologic changes. For example, after 10 days in hypoxia (9% oxygen), pulmonary artery pressure was significantly increased from 13 +/- 1 mmHg in normoxic controls (day 0, n = 6) to 26 +/- 0 mmHg (day 10, n = 4, P < 0.01). Right ventricular hypertrophy in hypoxic animals, presented as a ratio of right ventricle free wall weight to body weight, showed a significant increase from 0.054 +/- 0.004 (day 0) to 0.069 +/- 0.004 on day 10 (P < 0.05), while age-matched normoxic animals showed no changes in right ventricular weight (day 0 = 0.059, day 10 = 0.058; P > 0.05). Red blood cell count significantly increased over the same time period, from 5.9 +/- 0.1 (day 0) to 6.4 +/- 0.1 (day 10, P < 0.05), as did haematocrit, 48 +/- 0.7 (day 0) to 61 +/- 0.9 (day 10, P < 0.05), and haemoglobin, 16 +/- 0.2 (day 0) to 20 +/- 0.1 (day 10, P < 0.05). It is concluded that considerations for the well-being of the test animals (i.e. continuous water, ample food supplies, burrow-like hiding places, sanitation and protection from excess illumination) can easily be incorporated into a hypoxic chamber. The purpose of the present study was to explore modifications that may provide the animal with an optimized environment which will reduce anxiety and stress, as seen in their behaviour when inside the chambers, and to thoroughly characterize the morphologic and physiologic changes associated with chronic hypoxia which develop in a consistent time-related manner. PMID- 9350707 TI - Individual housing influences certain biochemical parameters in the rat. AB - Individual housing has been reported to modify animal behaviour. The present study compares the plasma levels of glucose, triglycerides and total cholesterol, weight, and food and water intake in two groups of female rats. Group A: 10 rats who remained grouped in two cages for 21 days; and Group B: 10 rats housed in two cages for 7 days, then isolated in individual cages from day 8 to day 15, and finally grouped together again for the last 7 days of the study. The results showed that the plasma values of glucose declined (P < 0.05) in the Group B rats when they had been returned to group condition (4.79 +/- 0.72 mM) than when they had been isolated (5.45 +/- 0.94 mM). Plasma triglyceride levels were lower (P < 0.05) in isolated rats (0.70 +/- 0.26 mM) than in any determination of the grouped rats. Group B: 1st week 1.21 +/- 0.21 mM, 3rd week 0.88 +/- 0.20 mM; and Group A: 1.22 +/- 0.20, 0.96 +/- 0.16, and 0.96 +/- 0.36 mM, in the first, second, and third week, respectively. There were no statistically significant differences in total cholesterol values as a function of the individual housing of animals. While there was no weight difference between the two groups of rats that could be ascribed to individual housing, there was a statistically significant increase (P < 0.05) in the food intake of isolated rats (17.5 +/- 3.2 g) with respect to values in the same Group B animals when they were housed together (1st week, 16.6 +/- 2.8 g; 3rd week, 16.8 +/- 3.1 g). These results therefore confirm that individual housing of female rats provoke variations in certain biochemical parameters, and that if this is not taken into account in performing different scientific studies, it could give rise to unreliable or even dubious results. PMID- 9350708 TI - An immunological assessment of group-housed rabbits. AB - Laboratory rabbits kept in barren 'traditional' cages tend to develop stereotypic behaviours and bone deformities. We have used an alternative regime, housing adult does as groups of four or five in floor pens (2.5-3 m2) supplied with hiding places and bedding. High- and low-ranking members of each group were identified, and their immunological status compared in terms of blood leucocyte function (chemiluminescence and mitogen tests), complement activity, and antibody production to soluble and cellular antigens. We found no evidence of immunosuppression, either in groups of a 'docile' breed (New Zealand White) or Dutch crosses. These results, together with the animals' general health and ease of handling, lead us to conclude that group-housed does are suitable for raising antisera and other purposes, provided that they are adequately monitored. PMID- 9350709 TI - Systemic mast cell disease in the Mongolian gerbil, Meriones unguiculatus: case report. AB - A case of spontaneous occurrence of systemic mast cell disease in a non-treated laboratory-reared gerbil is reported. This case corresponds to the form of systemic mastocytosis with bone marrow and visceral involvement associated with skin disease and displays clinically aggressive behaviour. The histopathological, histochemical and ultrastructural features are described. PMID- 9350710 TI - Hereditary cataract mouse with lens rupture: disputed priority. PMID- 9350712 TI - A model for the lateral penumbra in water of a 200-MeV proton beam devoted to clinical applications. AB - An experimental approach for modeling the lateral penumbra of a proton beam has been investigated. Measurements were made with a silicon diode in a water tank. Several geometrical configurations (phantom position, collimator-to-surface distance, collimator diameter, bolus thickness, air gap, etc.) and beam characteristics (range, modulation, etc.) have been studied. The results show that the lateral penumbra is almost independent of the beam modulation and the diameter of the collimator. The use of scaled variables for depth and penumbra allows us to represent the increase in penumbra with depth for any configuration with a second order polynomial function, provided that the penumbra at the entrance of the medium and at the depth of the range are known. PMID- 9350711 TI - Code of practice for brachytherapy physics: report of the AAPM Radiation Therapy Committee Task Group No. 56. American Association of Physicists in Medicine. AB - Recommendations of the American Association of Physicists in Medicine (AAPM) for the practice of brachytherapy physics are presented. These guidelines were prepared by a task group of the AAPM Radiation Therapy Committee and have been reviewed and approved by the AAPM Science Council. PMID- 9350713 TI - Evaluation of a cassette-screen-film combination for radiation therapy portal localization imaging with improved contrast. AB - A traditional limitation with radiation therapy portal images is low image contrast, due in part to the low attenuation of the exposing radiation by the tissues being imaged, and the contrast capabilities of the image receptor. We have developed, and have clinically evaluated, a cassette-screen-film combination for portal localization imaging, which features a copper front screen plus Gd2O2S:Tb fluorescent screens and a slow-speed, fine grain, film emulsion with inherently high contrast coated on both sides of a 7 mil Estar base. The film can be processed in a conventional rapid-process film processor. Sensitometric data indicate that the film contrast (average gradient) for the new combination is approximately 3.5 times higher than the conventional portal localization systems in current use. The new combination has been clinically compared with two conventional systems. The required monitor unit settings were found to be similar. Initial clinical results indicate portal images made with the new combination are superior to those obtained with the conventional combinations. The images have much higher contrast, subjective impressions of lower noise, show clearer definition of structures, and are much easier to read. PMID- 9350714 TI - Experimental determination of fluence correction factors at depths beyond dmax for a Farmer type cylindrical ionization chamber in clinical electron beams. AB - Recently, it has been recommended that electron beam calibrations be performed at a new reference depth [Burns et al., Med. Phys. 23, 383 (1996)] given by dref = 0.6R50-0.1 cm, where R50 is the depth of 50% depth dose. In order to calibrate electron beams at dref with a Farmer type cylindrical ionization chamber, the values of the perturbation correction factors Pwall and Pfl at dref are required. Using a parallel plate Holt chamber as a reference chamber, the product PwallPfl has been determined for a 6.1-mm-diameter PTW cylindrical ionization chamber at dref as a function of R50 of clinical electron beams (6 < or = nominal energy E < or = 22 MeV). Assuming that Pwall for the PTW chamber is unity in electron beams, the measured Pfl values ranged from 0.96 to 0.98 as the energy is increased. These results are in close agreement with recently reported calculated values. Determination of dref requires the knowledge of R50. A relation between I50 and R50 is given in the IAEA Protocol [TRS No. 277 (IAEA, Vienna, 1987), pp. 1-98] for broad beams at SSD = 100 cm. It has been shown experimentally that the equation R50 = 1.029 x I50-0.063 cm, derived by Ding et al. [Med. Phys. 22, 489 (1995)] from Monte Carlo simulations of realistic clinical electron beams, can be used satisfactorily to obtain R50 from I50, where I50 is the depth of 50% ionization. The largest difference between the measured value of R50 and that calculated by using the above equation has been found to be about 1 mm at 22 MeV. PMID- 9350715 TI - The CT's sample volume as an approximate, instrumental measure for density resolution in densitometry of the lung. AB - Ultimately CT-densitometry of the lung should give comparable results on all scanners. One prerequisite for this is the use of the same density resolution. Unfortunately, density resolution is impractical as a performance specifying parameter because it depends on the cellular material scanned. Therefore, another parameter that can be used for scanner and protocol characterization, and that does not depend on a special phantom, would be highly preferable. We investigated how well the CT's nominal sample volume (V), calculated from section thickness and in-plane spatial resolution as specified by the CT manufacturer, can serve as a simple measure, for density resolution. Six CT scanners were studied using foam and lung phantoms. On all scanners we observed for foam an approximately linear relation between density resolution and V-1/2. Density resolution on different scanners varied to some extent. These differences can be interpreted as being caused by deviations of the true sample volume from the nominal value: the 95% confidence interval runs for instance for V = 8 mm3 from 4.6 mm3 to 16.9 mm3. Acceptability of this spread depends on the consequences for parameters of clinical interest, like percentiles and pixel indexes. To evaluate this we used data from a previous patient study on the dependence of histogram parameters on sample volume. With these data it is found that large interscanner differences in histogram parameters are possible for small values of V, as used in thin-section densitometry. For larger values of V, as required for a more adequate density resolution, the differences are much smaller and probably acceptable when compared to other sources of variability in lung densitometry. In conclusion, for sections of at least 2 mm and smooth reconstruction filters, corresponding to V > or = 8 mm3, the CT's nominal sample volume might be used for interscanner and interprotocol comparison of density resolution. PMID- 9350717 TI - A method for simultaneous correction of spectrum hardening artifacts in CT images containing both bone and iodine. AB - A method is described capable of correcting artifacts in x-ray computer tomography (CT) images due to beam hardening in an arbitrary number of substances. The method works with reconstructed image data and does not require the original raw data. It is necessary to have an estimate of the spectrum of the incident x-ray beam. The method is similar to previously described iterative methods that correct artifacts induced by bones. Our implementation was designed to correct for hardening in both bone and iodine contrast agent. It is necessary to identify those regions of the image which contain bone and iodine. A central concept is that of effective density, which is the ratio of CT number of the substance to that of water. It is necessary to establish by a preliminary experiment the relationship between CT number and mass density of iodine or bone. From these data one estimates path integrals through soft tissue (water equivalent), bone, and iodine using a reprojection algorithm applied to the given image. Given this input, a key equation is solved numerically which provides a correction term to be subtracted from the reprojected data. This can be shown to eliminate the nonlinear terms in the projections due to beam hardening, assuming that the original density estimates were correct. In principle, the method can be repeated iteratively to improve the accuracy. However, in our experience using an image of a phantom containing iothalamate meglumine and K2HPO4, scanned using the Siemens Evolution electron beam tomography scanner, the quality of the corrected image was excellent and no further iteration is needed for the phantoms studied. More research is needed to implement the method on clinical scans. PMID- 9350716 TI - Relative speeds of Kodak computed radiography phosphors and screen-film systems. AB - Relative mAs values required to generate a constant plate readout signal for the Kodak Ektascan general purpose (GP-25) and high resolution (HR) photostimulable phosphors were measured as a function of x-ray beam quality and for a range of representative x-ray examinations. The signal intensity was determined from the exposure index (EI) generated during the read out of uniformly exposed phosphor imaging plates. These data were compared to the corresponding relative mAs values required to produce a constant film density of Lanex screen-film combinations with nominal speeds of 40, 400, and 600. The relative detection performance of the photostimulable phosphors generally decreased with increasing kVp and beam filtration. The relative response of GP-25 phosphors was independent of examination type, and modified by approximately 10% when scattered radiation was present. The HR phosphor was more efficient than a Lanex Single Fine extremity screen used with an EM-1 film. These relative response data will be useful for selecting the x-ray technique factors which minimize patient dose in x-ray examinations performed with photostimulable phosphors. PMID- 9350718 TI - Optimal pitch in spiral computed tomography. AB - The focus of this paper is to analytically optimize spiral/helical computed tomography (CT) protocols based on a simplified imaging model. Spiral CT was approximately modeled as follows: Using the half-scan raw data interpolation method, the variance of the spiral CT slice sensitivity profile is equal to the sum of squared detector collimation divided by 12 and squared table increment divided by 24. Image noise variance is inversely proportional to tube current and detector collimation. The maximum continuous scanning time is inversely proportional to tube current. Slice thickness, image noise, and signal-to-noise ratio were, respectively, optimized for a given scanning coverage, consistently resulting in pitch of square root of 2. To avoid longitudinal aliasing, at least 2-3 transverse slices should be reconstructed per collimation. When the simplified spiral CT model is valid and a scanning range specified, 1.4 pitch is required for optimal image quality. The method can be applied to more accurate spiral CT models. PMID- 9350719 TI - Measurement of the effective size of the input phosphor of an x-ray image intensifier. AB - A technique is described, using a mobile radiographic x-ray unit, for determining the effective size of the input phosphor of an image intensifier (I.I.), in particular for a unit with over-table x-ray tube, under-table II, and with the I.I. physically inaccessible. The techniques described also enable (1) locating the central axis of the I.I.; (2) the ratio, R, of the dimensions of the radiological image incident upon the front of the I.I. to the dimensions of the image on the monitor screen; and (3) the distance between the input phosphor and the table top, which is needed to determine the distance from focal spot to input phosphor. PMID- 9350720 TI - Suitability of laser stimulated TLD arrays as patient dose monitors in high dose x-ray imaging. AB - Skin entrance doses of patients undergoing interventional x-ray procedures are capable of causing skin damage and should be monitored routinely. Single TLD chips are not suitable because the location of maximum skin exposure cannot be predicted. Most photographic films are too sensitive at diagnostic x-ray energies for dosimetry, exhibit temporal changes in response, and require special packaging by the user. We have investigated the suitability of laser heated MgB4O7 TLDs in a polyimide binder in the range of 0.2-20 Gy. These are available in radioluscent arrays up to 30 x 30 cm for direct measurement of patient skin dose. Dose response was compared with a calibrated ion chamber dosimeter. Exposures were made at 60, 90, and 120 kVp, at low (fluoroscopy) and high (DSA) dose rates, and at different beam incidence angles. Longitudinal reproducibility and response to temperature changes during exposure were also checked. The dose response is linear below approximately 6 Gy where the slope starts to increase 2% per Gy. Errors were less than +/- 2% over a 0-80 degrees range of beam incidence angles; less than +/- 3% for both dose rate variations and kVp differences between 70 and 120 kVp. The response was unaffected by temperature changes between 20 and 37 degrees C. Reproducibility is current +/- 7%. MgB4O7 TLD arrays are suitable for patient dosimetry in high dose fluoroscopy procedures if appropriate calibrations are used. Uncertainty in skin dose measurement is less than 10%, which is substantially better than film dosimetry. PMID- 9350722 TI - Evaluation of three commercial enzyme-linked immunosorbent assays and two latex agglutination assays for diagnosis of primary Epstein-Barr virus infection. AB - Three commercially available enzyme-linked immunosorbent assays (ELISAs) from Gull, Biotest, and Behring (Enzygnost) and two latex agglutination tests for heterophile antibodies (Monolatex [Biotest] and Mono-Lex [Trinity Laboratories]) were evaluated for the diagnosis of primary Epstein-Barr virus (EBV) infection and EBV seropositivity. Two hundred fourteen consecutive samples from 197 patients with symptoms of primary EBV infection were analyzed by the five assays at a clinical microbiology laboratory. The samples were also analyzed independently by immunofluorescence methods at a reference laboratory. According to the reference methods, 37 patients (40 serum samples) had primary EBV infections, 120 patients (127 serum samples) had had past EBV infections, 33 patients (36 serum samples) were seronegative, and 7 patients (11 serum samples) exhibited atypical reactions. The respective sensitivities and specificities for the diagnosis of primary EBV infection were 95 and 100% for the Gull assays, 100 and 94% for the Biotest assays, and 100 and 89%, for the Enzygnost assays. The Monolatex and Mono-Lex methods showed similar sensitivities and specificities (78 to 85% and 100 to 99%, respectively) for the diagnosis of primary EBV infection. This study demonstrates the usefulness of commercially available assays for the rapid diagnosis of primary EBV infection, but also the importance of large-scale testing of routine samples before choosing an assay. PMID- 9350723 TI - Molecular diagnosis of bacterial endocarditis by broad-range PCR amplification and direct sequencing. AB - Broad-range PCR amplification of part of the 16S rRNA gene followed by single strand sequencing was applied to samples of 18 resected heart valves from patients with infective endocarditis. The PCR results were compared with those of cultures of valves and with those of previous blood cultures. For two patients there was agreement with the cultures of the valves; for nine patients there was agreement with the previous blood cultures, which were positive, while the cultures of the valves were negative; a Streptococcus sp. and Tropheryma whippelii each were found in one patient with negative cultures (valve and blood); for two patients the cultures of the valves as well as the PCR results were negative but the blood cultures were positive; for one patient amplification was inhibited; and for two patients the PCR results were positive but the amplicons could not be sequenced. It is concluded that broad-range PCR is a promising tool for patients with culture-negative endocarditis and allows the detection of rare, noncultivable organisms. PMID- 9350721 TI - Laboratory diagnosis of rickettsioses: current approaches to diagnosis of old and new rickettsial diseases. PMID- 9350724 TI - Quantification of cytomegalovirus DNA in peripheral blood leukocytes by a branched-DNA signal amplification assay. AB - Quantification of cytomegalovirus (CMV) DNA in blood may aid in the identification of patients at highest risk for developing CMV disease, the evaluation of new therapeutics, and the prompt recognition of drug-resistant CMV strains. A branched-DNA (bDNA) assay was developed for the reliable quantification of CMV DNA in peripheral blood leukocytes. The bDNA assay allowed for the highly specific and reproducible quantification of CMV DNA in clinical specimens. Furthermore, the bDNA assay was at least as sensitive as culture techniques and displayed a nearly 3 log10 dynamic range in quantification. Changes in CMV DNA levels measured by the bDNA assay in a human immunodeficiency virus-positive patient undergoing therapy were consistent with CMV culture, antigen, and genotype results and correlated with disease progression and resistance markers. The bDNA assay for the quantification of CMV DNA may provide a useful tool that can be used to aid physicians in monitoring disease progression, evaluating therapeutic regimens, and recognizing viral resistance and drug failure. PMID- 9350725 TI - Sequences of Pneumocystis carinii f. sp. hominis strains associated with recurrent pneumonia vary at multiple loci. AB - The sequences of the internal transcribed spacer (ITS) of Pneumocystis carinii f. sp. hominis strains from 7 of 15 AIDS patients were found to vary during discrete episodes of P. carinii pneumonia. Changes in the ITS sequence correlated with changes in the mitochondrial large-subunit rRNA sequence. The coincidence of changes in the sequences of the ITS, which is located in the nucleus, with changes in a mitochondrial gene excludes mutation as the cause of the genetic differences between P. carinii f. sp. hominis strains isolated during different episodes of P. carinii pneumonia and supports the hypothesis that recurrent P. carinii pneumonia is caused by reinfection rather than by reactivation of latent organisms. Thus, limiting the exposure of immunocompromised patients to P. carinii f. sp. hominis should help prevent P. carinii pneumonia. PMID- 9350726 TI - Rapid detection of Pneumocystis carinii in bronchoalveolar lavage specimens from human immunodeficiency virus-infected patients: use of a simple DNA extraction procedure and nested PCR. AB - We report on the development of a rapid nested PCR protocol for the detection of Pneumocystis carinii DNA in bronchoalveolar lavage (BAL) specimens in which the protocol included the use of a commercially available DNA extraction kit (GeneReleaser). GeneReleaser enabled us to obtain amplification-ready DNA within 20 min without requiring the purification of the DNA. The nested PCR was performed with the primers pAZ102-E, pAZ102-H, and pAZ102-L2 (A. E. Wakefield, F. J. Pixley, S. Banerji, K. Sinclair, R. F. Miller, E. R. Moxon, and J. M. Hopkin, Lancet 336:451-453, 1990.). Results were obtained in about 4 h with the adoption of denaturation, annealing, and extension steps shortened to 20 seconds. The sensitivity of the nested PCR was tested with a P. carinii cyst suspension and was found to be less than one cyst (one to eight nuclei). The detection limit was the same with the use of GeneReleaser or proteinase K-phenol chloroform for DNA extraction. The nested PCR assay was prospectively compared with staining with Giemsa and methenamine silver stains for the detection of P. carinii in 127 BAL samples from 105 human immunodeficiency virus-infected patients investigated for acute respiratory illness. Twenty-five BAL specimens (20%) were positive by staining and the nested PCR and 25 (20%) were negative by staining and positive by the nested PCR. These 25 BAL specimens with conflicting results were obtained from 23 patients, 82% of whom were receiving prophylactic therapy against P. carinii pneumonia (PCP). Only two patients were diagnosed with possible PCP. The final diagnosis was not PCP for 20 patients who were considered to be colonized or to have a low level of infection. This colonization is not of clinical importance but is of epidemiological importance. Our rapid, simple, and sensitive amplification protocol may be performed in clinical laboratories for the routine diagnosis of PCP with BAL specimens. PMID- 9350728 TI - Colonization of skin by Helcococcus kunzii. AB - In order to investigate the role of Helcococcus kunzii as a colonizer of skin and as a possible participant in diabetic foot ulcers, we used a selective medium to culture both lower- and upper-extremity skin from a study group of podiatry patients (60 diabetics and 60 nondiabetics) and a control group of 50 healthy volunteers. Although differences in colonization were not statistically significant, a trend toward higher colonization rates in the group of podiatry patients was noted. H. kunzii appears to preferentially colonize the skin of the feet, and while its pathogenic role in diabetic foot ulcers is difficult to establish, it may be a previously unrecognized component of the polymicrobial flora characteristically isolated from patients with these infections. PMID- 9350727 TI - Heterogeneity of BmpA (P39) among European isolates of Borrelia burgdorferi sensu lato and influence of interspecies variability on serodiagnosis. AB - The molecular and antigenic variabilities of BmpA (P39) among European isolates of Borrelia burgdorferi were analyzed. The bmpA sequences of 12 isolates representing all three species of B. burgdorferi sensu lato pathogenic for humans were amplified by PCR, cloned, and sequenced. The BmpA protein of Borrelia garinii is heterogeneous, with an amino acid sequence identity ranging from 91 to 97%, whereas the BmpA proteins of Borrelia afzelii and B. burgdorferi sensu stricto strains appear to be highly conserved (>98.5% intraspecies identity). The interspecies identities ranged from 86 to 92%. Cluster analysis of BmpA reflected the subdivision of B. burgdorferi sensu lato isolates into the three species as well as a considerable heterogeneity among B. garinii strains. The BmpA protein of each species of B. burgdorferi sensu lato was recombinantly expressed in Escherichia coli, purified, and used to generate monoclonal antibodies. Seven BmpA-specific antibodies were identified; six of them recognized conserved epitopes of all three species, whereas one was specific for BmpA of B. afzelii and B. garinii. A monoclonal antibody (H1141) recommended by the Centers for Disease Control and Prevention for use in the standardization of immunoblots showed strong reactivity with BmpA of B. burgdorferi sensu stricto but no or only weak reactivity with BmpA of B. garinii and B. afzelii, respectively. Sera from 86 European patients with Lyme borreliosis in different stages and 73 controls were tested in immunoglobulin G (IgG) and IgM immunoblots with the recombinant BmpA proteins of the three species, revealing specificities of 98.6 to 100%. IgM antibodies against recombinant BmpA were only rarely detected (1.1 to 8.1%). With the BmpA proteins of B. afzelii and B. garinii, sensitivities for the IgG test (sera from stages I to III) were 36.0 and 34.9%, respectively, in contrast to 13.9% with BmpA of B. burgdorferi sensu stricto. Therefore, we recommend that recombinant BmpA of B. afzelii or B. garinii should be used solely, or in addition to B. burgdorferi sensu stricto BmpA, in serodiagnostic tests for Lyme borreliosis in Europe. PMID- 9350729 TI - Heminested PCR assay for detection of six genotypes of rabies and rabies-related viruses. AB - A heminested reverse transcriptase PCR (hnRT-PCR) protocol which is rapid and sensitive for the detection of rabies virus and rabies-related viruses is described. Sixty isolates from six of the seven genotypes of rabies and rabies related viruses were screened successfully by hnRT-PCR and Southern blot hybridization. Of the 60 isolates, 93% (56 of 60) were positive by external PCR, while all isolates were detected by heminested PCR and Southern blot hybridization. We also report on a comparison of the sensitivity of the standard fluorescent-antibody test (FAT) for rabies antigen and that of hnRT-PCR for rabies viral RNA with degraded tissue infected with a genotype 1 virus. Results indicated that FAT failed to detect viral antigen in brain tissue that was incubated at 37 degrees C for greater than 72 h, while hnRT-PCR detected viral RNA in brain tissue that was incubated at 37 degrees C for 360 h. PMID- 9350730 TI - Molecular characterization of Mycobacterium avium complex isolates giving discordant results in AccuProbe tests by PCR-restriction enzyme analysis, 16S rRNA gene sequencing, and DT1-DT6 PCR. AB - Based on cultural and biochemical tests, a total of 84 strains (72 clinical and 12 environmental isolates from the Caribbean Isles, Europe, and the Indian subcontinent) were identified as members of the Mycobacterium avium complex (MAC). They were further characterized with MAC, M. avium, and M. intracellulare probes of the AccuProbe system, and this was followed by selective amplification of DT6 and DT1 sequences. Seventy isolates gave concordant results; 63 were identified as M. avium, 5 were identified as M. intracellulare, and 24 remained untypeable by both methods. Fourteen isolates gave discrepant results, as they were DT1 positive but gave negative results by the M. intracellulare AccuProbe test. Consequently, a detailed molecular analysis of all DT1-positive isolates (14 discrepant strains plus 5 M. intracellulare strains) was performed by PCR restriction analysis (PRA) of the hsp65 gene and 16S rRNA gene sequencing. The results confirmed the reported heterogeneity of M. intracellulare, as only 6 of 19 isolates (32%) gave PRA results compatible with published M. intracellulare profiles while the rest of the isolates were grouped in four previously unpublished profiles. 16S rRNA gene sequencing showed that only 8 of 19 isolates (42%) were related to M. intracellulare IWGMT 90247 (EMBL accession no. X88917), the rest being related to MCRO19 (EMBL accession no. X93030) and MIWGTMR10 (EMBL accession no. X88915). In conclusion, we have characterized a significant number of MAC isolates which were not identified by the AccuProbe test, PRA, or 16S rRNA sequencing. However, all of them were identifiable by DT1-DT6 PCR (they were DT6 negative and DT1 positive) and could be tentatively identified as M. intracellulare based on previously published observations. It is noteworthy that the majority of such isolates (14 of 19) were from the Indian subcontinent, with 12 of 14 being environmental isolates. Our study confirms the marked heterogeneity of M. intracellulare isolates and shows the utility of in-house DT1 PCR to detect this group of isolates, which would otherwise have been missed by the AccuProbe system in a routine clinical microbiology laboratory. PMID- 9350731 TI - Evaluation of BBL CHROMagar orientation medium for detection and presumptive identification of urinary tract pathogens. AB - The microbiological performance of BBL CHROMagar Orientation medium and CPS ID2 agar was compared to that of Columbia agar with 5% sheep blood and MacConkey agar without crystal violet for the enumeration and presumptive identification of bacteria responsible for urinary tract infections. Of a total of 658 clinical urine specimens, 118 specimens yielded no growth, 402 specimens yielded growth with cell counts of > or = 10(5) CFU/ml, and 138 specimens yielded growth with cell counts of < 10(5) CFU/ml. Of the specimens with cell counts of > or = 10(5) CFU/ml, 163 were pure cultures and 239 were mixed cultures. A total of 266 Escherichia coli organisms were isolated on both chromogenic media, 260 were isolated on blood agar, and 248 were isolated on MacConkey agar. One strain (0.4%) failed to develop the expected pink color on CHROMagar Orientation medium, and 23 strains (8.7%) failed to develop the expected pink color on CPS ID2 agar. Enterococci (CHROMagar Orientation medium, n = 266; CPS ID2 agar, n = 265) produced small blue-green colonies on both chromogenic media. Fifty of the mixed cultures contained enterococci that were detected only on the chromogenic media. The Klebsiella-Enterobacter-Serratia (KES) and the Proteus-Morganella-Providencia (PMP) groups could be identified on both chromogenic media. Of 66 isolates of the KES group, 63 grew with the expected color on CHROMagar Orientation medium and 58 of 64 isolates grew with the expected color on CPS ID2 agar. Other microorganisms required further identification. The use of chromogenic medium formulations offers a time-saving method for the reliable detection, enumeration, and presumptive identification of urinary tract pathogens. One of the greatest advantages of these media is the easy recognition of mixed cultures. PMID- 9350732 TI - Phenotypic and genotypic characterization of Vagococcus fluvialis, including strains isolated from human sources. AB - This study presents phenotypic and genotypic data for seven isolates of Vagococcus fluvialis, including four strains recovered from human clinical sources, one strain isolated from an environmental source, and two strains isolated from pigs. On the basis of phenotypic characteristics, most isolates were initially classified as "unidentified enterococci," because they resembled atypical arginine-negative enterococcal species. All seven strains as well as the type strain of V. fluvialis reacted with the AccuProbe Enterococcus genetic probe. The seven isolates had virtually indistinguishable whole-cell protein profiles that were similar to that of the V. fluvialis type strain and distinct from those of Enterococcus and Lactococcus species. DNA-DNA reassociation experiments confirmed that the strains were V. fluvialis. They were 71% or more related to the V. fluvialis type strain under optimum and stringent conditions, with 2.5% or less divergence within related sequences. All strains were susceptible to ampicillin, cefotaxime, trimethoprim-sulfamethoxazole, and vancomycin and were resistant to clindamycin, lomefloxacin, and ofloxacin. Strain to-strain variation was observed in relation to susceptibilities to 18 other antimicrobial agents. Chromosomal DNA was analyzed by pulsed-field gel electrophoresis (PFGE) after digestion with SmaI. Distinctive PFGE patterns were generated, suggesting the nonclonal nature of V. fluvialis strains. Although the number of strains was small, this report provides molecular characterization of V. fluvialis and the first evidence of a possible connection of this species with human infections. PMID- 9350734 TI - Human salmonellosis associated with exotic pets. AB - During the period from 1994 to 1996, an increase in the number of laboratory confirmed cases of human salmonellosis associated with exposure to exotic pets including iguanas, pet turtles, sugar gliders, and hedgehogs was observed in Canada. Pet turtle-associated salmonellosis was recognized as a serious public health problem in the 1960s and 1970s, and in February 1975 legislation banning the importation of turtles into Canada was enacted by Agriculture Canada. Reptile associated salmonellosis is once again being recognized as a resurgent disease. From 1993 to 1995, there were more than 20,000 laboratory-confirmed human cases of salmonellosis in Canada. The major source of Salmonella infection is food; however, an estimated 3 to 5% of all cases of salmonellosis in humans are associated with exposure to exotic pets. Among the isolates from these patients with salmonellosis, a variety of Salmonella serotypes were also associated with exotic pets and included the following: S. java, S. stanley, S. poona, S. jangwani, S. tilene, S. litchfield, S. manhattan, S. pomona, S. miami, S. rubislaw, S. marina subsp. IV, and S. wassenaar subsp. IV. PMID- 9350733 TI - Phenotypic and molecular characterization of three clinical isolates of Mycobacterium interjectum. AB - Introduction of molecular biology-based technology into an Australian mycobacterial reference laboratory has resulted in the identification of three isolates of Mycobacterium interjectum in the past 12 months. Conventional phenotypic methods failed to identify the species of these isolates, and high performance liquid chromatography found that only one of the three isolates had a mycolic acid pattern similar to that of the type strain. In contrast, all three isolates were rapidly identified as M. interjectum by 16S rRNA gene sequence analysis. Two isolates were recovered from the lymph nodes of children with cervical lymphadenitis, confirming the pathogenicity of this organism. However, the third isolate was obtained from the sputum of an elderly male with chronic lung disease without evidence of clinical or radiological progression, suggesting that isolation of M. interjectum should not imply disease. With the increasing use of molecular biology-based technology in mycobacterial laboratories, M. interjectum may be recognized more frequently as a pathogen or commensal organism. PMID- 9350735 TI - Inhibitory effects of polyoxyethylene stearate, PANTA, and neutral pH on growth of Mycobacterium genavense in BACTEC primary cultures. AB - We report on the influences of polyoxyethylene stearate (POES), PANTA, and pH on primary cultures of Mycobacterium genavense in BACTEC vials. As a model for primary cultures from tissue, seven different strains first isolated from AIDS patients (five from Switzerland and two from the United States) were inoculated into nude mice in order to obtain large amounts of bacilli to test different conditions simultaneously. Our results demonstrate that the size of the inoculum (10[6] acid-fast bacilli/vial), an acid pH (pH 6.0), and the absence of additives (POES and PANTA) significantly (P < 0.001) increased the probability of a successful culture in 1 month, considering growth index (GI) of > or =100 or a GI of > or =999 as criterion of success. In logistic regression analysis, all factors maintained a significant (P < 0.001) independent effect, and no interactions were observed between them. The best conditions for the primary cultures of M. genavense were the use of Middlebrook 7H12 medium at pH 6.0 without any additives. PMID- 9350736 TI - Quantification of human immunodeficiency virus type 1 RNA from dried plasma spots collected on filter paper. AB - To assess dried plasma spots (DPSs) as a source of material for virus quantification, human immunodeficiency virus type 1 (HIV-1) RNA levels were quantified in matched DPS and liquid plasma samples from 73 infected patients, including 5 neonates and 4 adult patients with acute HIV-1 infection. Quantifications were performed by commercially available assays (NASBA [nucleic acid sequence-based amplification] or Amplicor, or both). There was a strong correlation between HIV-1 RNA levels in plasma and DPSs. More importantly, there was no decline in HIV-1 RNA levels in DPSs stored for as long as 2 weeks at 20 degrees C. Similarly, storage of DPSs for 3 days at 37 degrees C resulted in no decrease in viral RNA levels. For patients with primary infection, the DPS method allowed for the measurement of RNA levels in plasma during the initial spike in the level of viremia and in the subsequent period of suppressed viral replication. DPS quantification was equally informative in the neonatal setting, with all five newborns showing HIV-1 RNA loads of greater than 4.991 log10 copies/ml. We conclude that the viral RNA levels in DPSs are equivalent to those measured in fresh-frozen plasma. The ease and economy of DPS sampling, the minute volumes required, and the unexpected stability of dried RNA suggest that the use of DPSs will be particularly valuable for small-volume neonatal samples and large, population-based studies in which cold storage and transportation present special problems, as is often the case in developing countries. The ability to measure viral changes during primary infection suggests that the method will be useful for assessing vaccine efficacy in large field trials. PMID- 9350737 TI - Detection of rifampin resistance by single-strand conformation polymorphism analysis of cerebrospinal fluid of patients with tuberculosis of the central nervous system. AB - Mutations in a 69-bp region of the rpoB gene of Mycobacterium tuberculosis are associated with rifampin resistance (Rif[r]). These have been detected with mycobacterial DNA extracted from bacterial suspensions or respiratory specimens that were acid-fast smear positive. We experimented with a strategy for the rapid detection of Rif(r) in cerebrospinal fluid (CSF) samples. The strategy involves the amplification of the 69-bp region of rpoB by means of PCR and the identification of nucleotide mutations by single-strand conformation polymorphism (SSCP) analysis of the amplification products. Sixty-five CSF specimens collected from 29 patients (19 patients were coinfected with human immunodeficiency virus) with culture or autopsy-confirmed (22 patients) or highly probable (7 patients) tuberculosis of the central nervous system (CNS-TB) were processed. Amplified products suitable for evaluation by SSCP analysis were obtained from 37 CSF specimens from 25 subjects (86.2%). PCR-SSCP of CSF correctly identified the rifampin susceptibility phenotype of isolates from all 17 patients for whom the results of susceptibility tests carried out with strains cultured from CSF or respiratory samples were available. Moreover, this assay revealed the rifampin susceptibility genotype of isolates from the eight patients (three patients with culture-confirmed CNS-TB and five patients in whom CNS-TB was highly probable) for whom no susceptibility test results were available; the PCR-SSCP data obtained for these patients were concordant with the outcome after a standard antituberculosis treatment. The evolution of a mutation in the rpoB gene was documented in a patient during the course of treatment. PCR-SSCP analysis of CSF seems to be an efficacious method of predicting Rif(r) and would reduce the time required for susceptibility testing from approximately 4 to 8 weeks to a few days. PMID- 9350738 TI - Molecular epidemiology of varicella-zoster virus in East London, England, between 1971 and 1995. AB - The molecular epidemiology of varicella-zoster virus in London, England, between 1971 and 1995 was examined by using two informative polymorphic markers, variable repeat region R5 and a BglI restriction site in gene 54. Viruses from 105 cases of chickenpox and 144 of zoster were typed. Two alleles of R5, A and B, were found at prevalences of 89 and 6%, respectively. No difference in allele frequency between the zoster and chickenpox cases was found, and no change in the frequencies of these alleles was observed to occur over time. By contrast, a BglI restriction site (BglI+) was found with increasing frequency over time among cases of varicella (P < 0.005) and, to a lesser extent, cases of zoster. The BglI+ polymorphism was strongly associated (P < 0.0005) with zoster in subjects who had immigrated to the United Kingdom from countries with low adult immunity to varicella (LAIV). Sixty-three percent of the subjects with zoster who had emigrated from countries with LAIV carried the BglI+ virus, in contrast to 10% of adults who had grown up in countries with high adult immunity to varicella. The significance of these data, in view of the changing epidemiology of chickenpox, is discussed. PMID- 9350739 TI - Discrimination of Campylobacter jejuni isolates by fla gene sequencing. AB - Comparison of the entire coding sequence of flaA (1,764 nucleotides) from 15 isolates of Campylobacter jejuni showed two regions of high variability, one region approximately from base positions 700 to 1,450 and a short variable region (SVR) from base positions 450 to 600. Parsimony analysis of the SVR sequences yielded a dendrogram similar to that which was derived by analysis of the entire gene. PCR was used to generate templates, and the SVR was sequenced with primers constructed to hybridize to conserved flanking sequences. The SVRs of 22 isolates of C. jejuni from four outbreaks that have been well characterized and a larger panel of isolates from three additional outbreaks were sequenced. Analysis of the nucleotide sequences produced results that grouped the isolates very similarly to other subtyping techniques. Sequence data were also generated for isolates from three additional outbreaks. Categorizing the isolates by fla SVR DNA sequence placed them in epidemiologically relevant groups. Sequence analysis of the C. jejuni flaA SVR may be a useful tool for epidemiologic investigations and could complement or replace serotyping and other subtyping methods. PMID- 9350740 TI - Report of spores of Henneguya salminicola (Myxozoa) in human stool specimens: possible source of confusion with human spermatozoa. AB - The spores of Henneguya salminicola, a common tissue parasite of salmonid fishes in the northern hemisphere, were observed in stool specimens from two different patients with diarrhea. The spores' superficial resemblance to human spermatozoa resulted, in one instance, in an incorrect report, leading to suspicion of sexual abuse. H. salminicola spores and human spermatozoa can be differentiated on the basis of size, morphology, and staining characteristics. Laboratory personnel who perform microscopic examinations of stool specimens for ova and parasites should be aware that spores of H. salminicola may be seen from time to time. PMID- 9350742 TI - Recent emergence of new variants of Yersinia pestis in Madagascar. AB - Yersinia pestis, the causative agent of plague, has been responsible for at least three pandemics. During the last pandemic, which started in Hong Kong in 1894, the microorganism colonized new, previously unscathed geographical areas where it has become well established. The aim of this longitudinal study was to investigate the genetic stability of Y. pestis strains introduced into a new environment just under a century ago and to follow the epidemiology of any new genetic variant detected. In the present study, 187 strains of Y. pestis isolated between 1939 and 1996 from different regions of Madagascar and responsible mainly for human cases of bubonic and pneumonic plague were studied. Our principal genotyping method was rRNA gene profiling (ribotyping), which has previously been shown to be an effective scheme for typing Y. pestis strains of different geographical origins. We report that all studied Y. pestis strains isolated in Madagascar before 1982 were of classical ribotype B, the ribotype attributed to the Y. pestis clone that spread around the world during the third pandemic. In 1982, 1983, and 1994, strains with new ribotypes, designated R, Q, and T, respectively, were isolated on the high-plateau region of the island. Analysis of other genotypic traits such as the NotI genomic restriction profiles and the EcoRV plasmid restriction profiles revealed that the new variants could also be distinguished by specific genomic and/or plasmid profiles. A follow-up of these new variants indicated that strains of ribotypes Q and R have become well established in their ecosystem and have a tendency to spread to new geographical areas and supplant the original classical strain. PMID- 9350741 TI - Distribution of Acinetobacter species on human skin: comparison of phenotypic and genotypic identification methods. AB - At least 19 genomic species are recognized as constituting the genus Acinetobacter. However, little is known about the natural reservoirs of the various members of the genus. An epidemiological study was therefore performed to investigate the colonization with Acinetobacter spp. of the skin and mucous membranes of 40 patients hospitalized in a cardiology ward and 40 healthy controls. Single samples were obtained once from each of nine different body sites, i.e., forehead, ear, nose, throat, axilla, hand, groin, perineum, and toe web. Identification of Acinetobacter isolates was achieved by using phenotypic properties and was compared to identification by amplified ribosomal DNA restriction analysis. Selected isolates were further investigated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis, ribotyping, and DNA-DNA hybridization. Plasmid profile analysis was used for epidemiological typing. Thirty patients (75%) and 17 controls (42.5%) were found to be colonized with Acinetobacter spp., and the colonization rates of patients increased during their hospital stay. The most frequently isolated species were Acinetobacter lwoffii (47%), A. johnsonii (21%), A. radioresistens (12%), and DNA group 3 (11%). In contrast, A. baumannii and DNA group 13TU, the most important nosocomial Acinetobacter spp., were found only rarely on human skin (0.5 and 1%, respectively) and their natural habitat remains to be defined. A good correlation between phenotypic and genotypic methods for identification of Acinetobacter spp. was observed, and only two isolates could not be assigned to any of the known DNA groups. PMID- 9350743 TI - Genotype-specific RNA probes for direct identification of wild polioviruses by blot hybridization. AB - We have developed RNA probes for the direct identification of wild poliovirus isolates by blot hybridization. The probes are complementary to sequences of the first 30 to 32 codons of VP1, which evolve more extensively (approximately 1.5 fold) than the rest of VP1. To illustrate our general approach, we describe the design of probes specific to each of four major genotypes recently endemic (1981 to 1991) to the Americas: Andean type 1, Brazil type 1, Brazil type 3, and Central America-Mexico type 3. A wild isolate of each genotype was selected according to molecular and epidemiologic criteria to be representative of the principal lineages in circulation. Variable VP1 sequences of the representative isolates were amplified by the reverse transcriptase PCR and were inserted into a plasmid vector containing a T7 promoter. The in vitro transcripts, labeled with digoxigenin, served as probes. These formed stable hybrids only with RNAs of isolates of the corresponding genotypes. Hybrids were detected by a sensitive chemiluminescence assay, capable under normal diagnostic conditions of detecting specific wild poliovirus sequences in samples containing up to a 100-fold excess of Sabin vaccine strain-related sequences of the same serotype. PMID- 9350744 TI - Antigenic lipopolysaccharide components of Legionella pneumophila recognized by monoclonal antibodies: possibilities and limitations for division of the species into serogroups. AB - Legionella pneumophila accounts for the majority of cases of Legionnaires' disease. By using rabbit antisera, the species has been divided into 14 numbered and 1 unnumbered serogroups. To recognize the antigenic diversity of the lipopolysaccharide (LPS) responsible for this classification, the Dresden Legionella LPS MAb panel, containing 98 monoclonal antibodies (MAbs), was created. Each serogroup reference strain possesses at least one specific epitope not found on any other reference strain and therefore designated the serogroup specific epitope. When the appropriate MAbs were used for serotyping of 1,064 human and environmental isolates, 1,045 (98%) could be placed into the known serogroups. In most cases (97%), this was in agreement with the polyclonal typing. Of the 29 isolates that showed strong cross-reactivities with the rabbit antiserum panel, 11 could be typed easily by MAbs; for the remaining 18, however, only serogroup-cross-reactive epitopes could be determined. Below the serogroup level, monoclonal subtypes were found for 11 serogroups. Altogether, the Dresden Legionella LPS MAb panel was able to divide the 1,064 isolates tested into 64 phenons, indicating its usefulness for both serogrouping and subgrouping of L. pneumophila strains. In order to compare the identities of patient and environmental isolates, testing their reactivity with MAbs should be the first step, especially if large numbers of colonies are to be typed. Only in cases of identical patterns are the more time consuming and expensive genetic fingerprints necessary. Moreover, the MAbs can also be used for specific antigen detection in respiratory specimens on the serogroup or subgroup level. PMID- 9350745 TI - Performance of the Amplicor human immunodeficiency virus type 1 PCR and analysis of specimens with false-negative results. AB - Over a 4-year period, the Roche Amplicor kit was used in a United Kingdom reference laboratory for the detection or confirmation of human immunodeficiency virus (HIV) type 1 infection, particularly in infants born to HIV-infected mothers. Of 408 specimens from adults and older children tested, the 122 seronegative specimens were all Amplicor negative. Of the 286 seropositive specimens, 268 were Amplicor positive. On the basis of these results, the Amplicor assay has a specificity of 100% and a sensitivity of 93.7%. In addition, for 247 specimens from infants and young children, serological results may not have been diagnostic because of placental transfer of maternal antibodies. Forty eight were Amplicor positive, and of the 199 Amplicor-negative specimens, 19 were assumed to be false negative on the basis of clinical data, serological markers (including p24 antigen), and/or results for previous or follow-up specimens. This represents a sensitivity of 75% for the Amplicor test for specimens from patients under 2 years of age. Of these 37 false-negative specimens plus 2 specimens from other laboratories, 31 could be characterized by amplifying extracted material from them by an in-house nested gag PCR spanning the Amplicor target region. The amplicons were sequenced and found to represent subtypes A (35.5%), B (22.6%), C (22.6%), D (16.1%), and G (3.2%). False-negative results by the Amplicor assay may be ascribed to low-target copy number, the physical behavior of one primer (SK462), and sequence variation in the target region of the other primer (SK431). PMID- 9350746 TI - Use of multiplex PCR for simultaneous detection of four bacterial species in middle ear effusions. AB - A multiplex PCR procedure was developed for the simultaneous detection of Alloiococcus otitidis, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae in middle ear effusions (MEEs) from patients with chronic otitis media with effusion. The bacterial 16S rRNA gene was chosen as the target, and the procedure used one common lower primer and four species-specific upper primers. The reaction was optimized by changing the primer concentrations to yield equal amounts of amplification products. The specificity of the reaction was verified with various bacterial species found in the nasopharynx. The performance of the procedure was examined with 25 MEE specimens, and the results were compared to those obtained by conventional culture methods. A detection level of 10 bacterial cells/reaction for each of the study organisms was achieved. By conventional culture methods, 8 (32%) of the specimens showed growth of one of the study organisms. In contrast, 21 (84%) of the specimens tested positive by the multiplex PCR. None of the culture-positive specimens were PCR negative, whereas three (12%) of the PCR-positive specimens tested positive for two of the four study organisms. Thus, the multiplex PCR method improves the detection rate significantly compared to that of the conventional culture method. PMID- 9350747 TI - Use of pulsed-field gel electrophoresis to determine genomic diversity in strains of Helicobacter hepaticus from geographically distant locations. AB - In 1992 a helical microorganism associated with chronic active hepatitis and a high incidence of hepatocellular tumors was identified in the hepatic parenchyma of A/JCr mice. By using biochemical tests, phenotypic characterization, and 16S rRNA gene sequence analysis, the organism was classified as a novel Helicobacter species and named Helicobacter hepaticus. Recent surveys completed in our laboratory indicate that H. hepaticus is widespread in academic and commercial mouse colonies. The aim of this study was to examine the H. hepaticus genome by pulsed-field gel electrophoresis (PFGE) to determine the degree of genomic variation and genomic size. This technique has been used to identify significant genomic diversity among strains of Helicobacter pylori and to demonstrate only slight genomic diversity among strains of Helicobacter mustelae. Genomic DNAs from 11 isolates of H. hepaticus from the United States, Germany, France, and The Netherlands were subjected to PFGE after digestion with SmaI. Isolates from three independent sources within the United States had very similar PFGE patterns, suggesting that the genomic DNAs of these isolates are conserved. Genomic DNA isolated from a fourth source within the United States had a PFGE pattern different from those of the other U.S. isolates. Isolates obtained from Germany, France, and The Netherlands had PFGE patterns that differed markedly from those of the U.S. isolates and from one another. The use of DNA fingerprinting may be useful in subsequent epidemiological studies of H. hepaticus when the source and method of spread of this murine pathogen need to be ascertained. By PFGE, the genomic size of H. hepaticus is estimated to be roughly 1.3 Mb, which compares to 1.67 Mb for H. pylori and 1.7 Mb for H. mustelae. PMID- 9350748 TI - Frequency and natural history of rhinovirus infections in adults during autumn. AB - Human rhinovirus (HRV) accounts for a significant portion of common-cold illness, with the peak incidence being in the early fall. Three hundred forty-six adults who had self-diagnosed colds of 48 h or less were enrolled in a study during September and October 1994 to determine the frequency and clinical course of HRV infections. Nasal wash specimens for viral culture and reverse transcription-PCR (RT-PCR) for HRV RNA and human coronavirus OC43 and 229E RNA detection were collected on enrollment, and participants recorded their symptoms twice daily for 14 days. Middle ear pressure (MEP) was measured with a digital tympanometer on days 1 and 7. Picornaviruses (224 HRV and 7 enterovirus isolates) were detected by culture in 67% (231 of 346) of the subjects. Among 114 samples negative by culture, HRV was detected by RT-PCR in 52 (46%) for an overall picornavirus infection rate of 82% (283 of 346 subjects). Among the remaining 62 negative samples, human coronavirus RNA was detected by RT-PCR in 5 patients, so that 288 (83%) of patients had documented viral infection. The first symptom noticed most often was sore throat (40%) in HRV culture- or PCR-positive patients and stuffy nose in HRV-negative patients (27%). No differences in symptom scores over time or in the presence of individual symptoms were noted between groups. The median duration of the cold episodes was 11 days in HRV culture-positive patients, 9.5 days in HRV RT-PCR-positive patients, and 11.5 days in HRV-negative patients. On enrollment, abnormal MEPs (< or = -100 or > or = +100 mm of H2O) were found for 21% of HRV culture-positive patients, 14% of HRV RT-PCR-positive patients, and 10% of HRV-negative patients. No important differences in the clinical course of HRV culture-positive, HRV culture-negative and RT-PCR-positive, or HRV-negative colds were found. These results represent the highest frequency of virologically confirmed natural colds to date and document the importance of rhinoviruses as the cause of colds during fall months. PMID- 9350749 TI - Molecular evidence and clinical significance of herpesvirus coinfection in the central nervous system. AB - A total of 60 cerebrospinal fluid (CSF) specimens from patients manifesting symptoms resembling viral central nervous system (CNS) disease were examined for the presence of herpes simplex virus (HSV), human herpesvirus 6 (HHV-6), Epstein Barr virus (EBV), cytomegalovirus, varicella-zoster virus, Borrelia burgdorferi, and Tropheryma whippelii DNA by PCR. Of 30 specimens which were selected on the basis of HSV DNA positivity, 2 were concomitantly positive for HHV-6 DNA and 1 was positive for EBV DNA. In the three specimens positive for more than one herpesvirus, amplicons generated with virus-specific primer sets hybridized specifically to the corresponding virus-specific probe. Sequence analysis of the two amplified DNA fragments demonstrated that they were derived from distinct herpesviruses. Of 22 patients with clinically diagnosed encephalitis, 2 of 3 patients coinfected with HSV and HHV-6 died, compared to 1 of 19 (5%) patients infected with only HSV. Of 30 CSF specimens that were negative for HSV DNA, EBV DNA was detected in one sample. These data indicated the presence of DNA specific for two distinct herpesviruses in the same CSF specimen, providing molecular evidence that coinfection with this group of viruses may occur in the CNS. PMID- 9350752 TI - Seroprevalence of Bartonella henselae infection and correlation with disease status in cats in Switzerland. AB - The prevalence of infection with Bartonella henselae was investigated in cats from different areas of Switzerland. Serum samples of 728 cats were examined for antibodies to B. henselae by immunofluorescent antibody testing, and the results were analyzed with a view to a possible correlation between a positive titer and signalment, clinical signs, infection with feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), feline coronavirus (FCoV), or feline spumavirus (FeSFV), and the living environments of the cats. The seroprevalence in all cats was 8.3%. No significantly different prevalence was found in sick versus healthy cats (9.2 versus 7.2%); however, in sick cats seropositive for B. henselae, there was an increased frequency of stomatitis and a variety of diseases of the kidneys and the urinary tract. There was an increased prevalence of B. henselae in cats positive for FCoV (P = 0.0185) or FeSFV (P = 0.0235) and no statistically significant increased prevalence in cats infected with FeLV or FIV. There was no correlation between a positive titer and sex or breed. The same prevalence of B. henselae antibodies was found in cats with and without access to the outdoors and in cats from single- and multicat households. The seroprevalence was increased in cats living south of the Alps (12.1%); however, this difference was not significant (P = 0.0616). PMID- 9350750 TI - Laboratory diagnosis of central nervous system infections with herpes simplex virus by PCR performed with cerebrospinal fluid specimens. AB - Until recently, the laboratory diagnosis of central nervous system (CNS) infections with herpes simplex virus (HSV) has been limited by poor sensitivity and/or specificity. We assessed the diagnostic utility of PCR for detection of HSV in over 2,100 specimens referred to the Mayo Clinic from August 1993 to May 1996. DNA extracted from cerebrospinal fluid (CSF) samples with IsoQuick was amplified by PCR with oligonucleotide primers directed to the DNA polymerase gene of HSV, yielding a 290-bp amplicon. HSV DNA was detected in 150 (135 by gel electrophoresis, 15 by Southern blotting only) of 2,106 (7.1%) specimens. PCR positive CNS disease occurred in patients ranging in age from 13 days to 89 years; 59% of the cases occurred in patients between the ages of 30 and 69, and 21 (14%) of the patients were infants. Genotype analysis was not routinely performed; however, amplification of a 335-bp product within the thymidine kinase gene of HSV revealed 13 positions within a span of 80 nucleotides that accurately identified the two serotypes of the virus according to 14 reference strains. We conclude that PCR detection of HSV DNA in CSF specimens should be considered an emerging "gold standard" for the laboratory diagnosis of CNS infections with this virus. PMID- 9350751 TI - Comparison of a photometric method with standardized methods of antifungal susceptibility testing of yeasts. AB - We determined the fluconazole MICs for 101 clinical isolates of Candida and Cryptococcus neoformans using the macro- and microdilution methods recommended by the National Committee for Clinical Laboratory Standards. We compared the MICs obtained by these methods with those obtained by a photometric assay that quantified the reduction of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) by viable fungi. The MIC determined by this method was defined as the highest fluconazole concentration associated with the first precipitous drop in optical density. For Candida, both the MTT and the microdilution methods demonstrated excellent agreement with the standard macrodilution method. The MTT method, however, generated MICs at 24 h that were comparable to those generated by the standard macrodilution method, whereas the microdilution method required 48 h. For C. neoformans, the levels of agreement between the MICs determined by the MTT and microdilution methods after 48 h and those determined by the standard 72-h macrodilution method were 94% (29 of 31) and 94% (29 of 31), respectively. The MTT method therefore provided results comparable to those of currently recommended methods and had the advantages of a more rapid turnaround time and potential adaptability to use as an automated system. Furthermore, the MICs determined by the MTT method were determined photometrically, thereby eliminating reader bias. PMID- 9350753 TI - Effects of specimen collection, processing, and storage conditions on stability of human immunodeficiency virus type 1 RNA levels in plasma. AB - To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 (HIV-1) viral load testing, plasma HIV-1 RNA levels were quantitated with the NASBA HIV-1 RNA QT System from blood specimens that were collected, processed, and stored under a variety of conditions that might have affected HIV-1 RNA stability. We determined that when whole blood was processed within 2 h of specimen collection the levels of HIV-1 RNA detected in EDTA-, heparin-, and acid citrate dextrose (ACD)-anticoagulated plasma samples were comparable. The levels of HIV-1 RNA in serum specimens (mean = 4.126 log units) were significantly lower (P < 0.01) than the levels in corresponding plasma samples (mean = 4.501 log units). One cycle of freeze-thaw (-70 degrees C) did not significantly reduce the level of HIV-1 RNA detected in EDTA-, heparin-, or ACD-anticoagulated plasmas. The EDTA-anticoagulated plasmas showed the smallest decrease in HIV-1 RNA copies (0.050 log units). HIV-1 RNA levels decreased over a 6-month time period in serum as well as in EDTA-, ACD-, and heparin-anticoagulated plasmas stored at -70 degrees C. However, the only significant decreases were for serum (mean decrease = 0.317 log units) and heparin-anticoagulated samples (mean decrease = 0.384 log units). A comparison of the levels of HIV-1 RNA in cell-free plasma collected in VACUTAINER EDTA Plasma Preparation Tubes and in standard VACUTAINER EDTA tubes determined that HIV-1 RNA levels were stable for up to 30 h after collection when stored at either room temperature (mean standard deviation [SD] = +/- 0.101 log units) or at 4 degrees C (mean SD = +/- 0.102 log units) as cell-free plasma or as EDTA-anticoagulated whole blood (mean SD = +/- 0.109 log units). These data indicate that EDTA anticoagulated plasma is the most suitable and stable matrix for HIV-1 RNA quantitation. PMID- 9350754 TI - Molecular epidemiology of Ornithobacterium rhinotracheale. AB - Ornithobacterium rhinotracheale is a recently described gram-negative rod-shaped bacterium associated with respiratory tract infections in poultry. In order to determine the molecular epidemiology of this bacterium, we characterized 55 O. rhinotracheale isolates from eight countries on four continents by multilocus enzyme electrophoresis (MLEE), repetitive sequence based-PCR (rep-PCR), and 16S rRNA gene sequencing. MLEE discriminated the O. rhinotracheale isolates into six electrophoretic types (ETs), of which only three ETs were recovered from domesticated poultry. The 16S rRNA gene sequence and rep-PCR analyses confirmed the results obtained by MLEE and indicated limited heterogeneity among isolates of O. rhinotracheale recovered from poultry. Taken together, the results of our analysis demonstrate that the majority of O. rhinotracheale isolates recovered from domesticated poultry throughout the world are represented by a small group of closely related clones and suggest that the bacterium was recently introduced to domesticated poultry from wild bird populations. PMID- 9350756 TI - An outbreak of listeriosis suspected to have been caused by rainbow trout. AB - An outbreak of listeriosis in Sweden, consisting of nine cases, was investigated by means of molecular typing of strains from patients and strains isolated from suspected foodstuffs, together with interviews of the patients. Listeria monocytogenes was isolated from six of the patients, and all isolates were of the same clonal type. This clonal type was also isolated from a "gravad" rainbow trout, made by producer Y, found in the refrigerator of one of the patients. Unopened packages obtained from producer Y were also found to contain the same clonal type of L. monocytogenes. Based on the interview results and the bacteriological typing, we suspect that at least six of the nine cases were caused by gravad or cold-smoked rainbow trout made by producer Y. To our knowledge, this is the first rainbow trout-borne outbreak of listeriosis ever reported. PMID- 9350755 TI - Vaginal carriage of enterotoxigenic Bacteroides fragilis in pregnant women. AB - Bacteroides fragilis is an anaerobic bacterial species that is involved in gynecological infections and pathology. The incidence of vaginal carriage is largely unknown, and in order to study this, 120 pregnant women attending a general hospital for delivery were examined. Cultures were positive for eight of these women (6.6%). Interestingly, potential clonal relatedness could be demonstrated among several of the nonenterotoxigenic B. fragilis strains. Among the strains, only one produced metalloprotease enterotoxin. The presence of the gene for the metalloprotease, giving rise to the pathogenic effect on cultured eukaryotic HT29/C1 cells, was confirmed by a newly designed specific PCR assay. The enterotoxigenic B. fragilis (ETBF) strain was analyzed with the help of arbitrarily primed PCR (AP-PCR) and PCR-mediated ribotyping. The ETBF strain was shown to be genetically different compared to several other strains obtained from diverse sources. Our data indicate a relatively high vaginal B. fragilis carriage rate among pregnant women in Warsaw, Poland. Although neither ETBF nor B. fragilis colonization presented a clinical problem, the possible genetic relatedness among the colonizing B. fragilis strains indicates the need for additional research in the field of ETBF transmission and molecular epidemiology. PMID- 9350757 TI - Phylogenetic analyses of Chlamydia psittaci strains from birds based on 16S rRNA gene sequence. AB - The nucleotide sequences of 16S ribosomal DNA (rDNA) were determined for 39 strains of Chlamydia psittaci (34 from birds and 5 from mammals) and for 4 Chlamydia pecorum strains. The sequences were compared phylogenetically with the gene sequences of nine Chlamydia strains (covering four species of the genus) retrieved from nucleotide databases. In the neighbor-joining tree, C. psittaci strains were more closely related to each other than to the other Chlamydia species, although a feline pneumonitis strain was distinct (983 to 98.6% similarity to other strains) and appeared to form the deepest subline within the species of C. psittaci (bootstrap value, 99%). The other strains of C. psittaci exhibiting similarity values of more than 99% were branched into several subgroups. Two pigeon strains and one turkey strain formed a distinct clade recovered in 97% of the bootstrapped trees. The other pigeon strains seemed to be distinct from the strains from psittacine birds, with 88% of bootstrap value. In the cluster of psittacine strains, three parakeet strains and an ovine abortion strain exhibited a specific association (level of sequence similarity, 99.9% or more; bootstrap value, 95%). These suggest that at least four groups of strains exist within the species C. psittaci. The 16S rDNA sequence is a valuable phylogenetic marker for the taxonomy of chlamydiae, and its analysis is a reliable tool for identification of the organisms. PMID- 9350759 TI - Growth and morphological transformations of Helicobacter pylori in broth media. AB - Helicobacter pylori, a cause of peptic ulcer disease and certain types of gastric cancers, has usually been cultured on diverse agar-based media, resulting in a requirement for 2 to 4 days of growth at 37 degrees C. We have developed a novel broth medium consisting of a base medium supplemented with 2% newborn calf serum, Mg2+, Cu2+, Fe2+, Zn2+, Mn2+, and 1 mg of lysed human erythrocytes per ml. This medium supports rapid growth of H. pylori, with a doubling time of about 50 min. Optimal growth was obtained in a pH range higher than that supporting most other gram-negative bacteria (at pH 8.5). H. pylori cultured in this supplemented broth retains the spiral morphology seen in both histological sections and cultures from agar-based media and also retains a high urease activity. After 18 h in this broth, H. pylori transforms to a coccal form with a complete loss of urease activity. Previously these cocci have been reported to be senescent, since they could not be subcultured on agar medium. Our experiments suggest that some of the cocci can revert back to the spiral morphology with full recovery of urease activity when subcultured in fresh microaerobic broth medium. PMID- 9350758 TI - Culture of the causative organism of donovanosis (Calymmatobacterium granulomatis) in HEp-2 cells. AB - We report successful culture of Calymmatobacterium granulomatis by standard cell culture methods. Swabs were obtained from lesions in three patients with a clinical diagnosis of donovanosis. For two patients, there was histological confirmation of the disease (i.e., the presence of Donovan bodies in Giemsa stained smears). Specimens were inoculated onto cycloheximide-treated HEp-2 cell monolayers in RPMI 1640 medium (supplemented with fetal calf serum, NaHCO3, vancomycin hydrochloride, and benzylpenicillin). At 48 h, organisms resembling Donovan bodies were identified in monolayer cultures from all three specimens. The organisms appeared as pleomorphic bacilli with characteristic bipolar staining and "safety pin" appearance. Using a PCR designed to differentiate C. granulomatis from the Klebsiella species (which have a high degree of molecular homology), we were able to demonstrate that the cultured organisms produced a PCR product identical to that obtained from the original swab specimens. It is now possible to test in vitro susceptibility of C. granulomatis to antibiotics and to provide a ready source of DNA and antigenic material to enable the development of serological tests and, possibly in the future, a vaccine. PMID- 9350760 TI - Decreased capacity for type-specific-antigen synthesis accounts for high prevalence of nontypeable strains of group B streptococci in Mexico. AB - The low incidence of group B streptococcal (GBS) invasive neonatal disease in Mexico has been attributed to the low prevalence of serotype III strains, a major serotype in developed countries. In addition, nontypeable strains account for 12% of the isolates in Mexico and < 1% of the isolates in the United States. In this study, 57 GBS isolates (28 nontypeable by the Lancefield procedure) from carrier and infected neonates and women from Mexico were also examined for the presence of type-specific antigen by an enzymatic procedure using N-acetylmuramidase digestion of the cell wall to release soluble type-specific antigen. Of the 28 nontypeable strains from Mexico, 23 were typeable by the enzyme extraction procedure, with serotype III being the predominant serotype in invasive disease. These results suggest that nontypeable isolates of GBS should be further examined by the enzymatic extraction procedure to determine the presence of type-specific antigen. Furthermore, these limited results suggest that serotype III is likely a major serotype in invasive disease also in Mexico. PMID- 9350762 TI - Evaluation of performances of three DNA enzyme immunoassays for detection of Helicobacter pylori PCR products from biopsy specimens. AB - PCR is recognized as a promising method for the detection of Helicobacter pylori in gastric biopsy specimens. However, detection of PCR products by gel electrophoresis is difficult to implement in routine clinical laboratories. The aim of this study was to compare three new DNA enzyme immunoassays with the standard method in their ability to detect PCR products. The three assays were based on the amplification of a fragment of the ureC gene of H. pylori and a colorimetric hybridization assay. The first assay (GEN-ETI-K DNA enzyme immunoassay; Sorin, Sallugia, Italy) was based on the hybridization of amplified DNA with a probe bound in microtiter wells and detected with labelled anti-DNA antibody. The second assay (Pylori-prob; Biocode, Sclessin, Belgium) comprised a solid-phase sandwich hybridization system with a specific biotinylated probe being used for detection. Finally, the third assay (PCR enzyme-linked immunosorbent assay; Boehringer, Mannheim, Germany) was based on the hybridization of amplified DNA labelled with digoxigenin as a probe (used as a coating in microtiter wells) and detected with antidigoxigenin-peroxidase as conjugate. The sensitivity of the colorimetric assay was evaluated by using amplification products from PCR assays performed on several 10-fold dilutions of DNA from H. pylori CIP 101260, and the specificity was assessed with different urease-positive bacteria. Biopsy specimens from 199 patients were tested; 106 were classified as H. pylori positive, and 93 were classified as H. pylori negative by culture and/or histological examination as the "gold standard." The receiving operating characteristic curve was used to determine the best cutoff point for each assay. The detection of PCR products by colorimetric hybridization increases the sensitivity up to 100-fold compared to that with gel electrophoresis. The results are rapid (4 h) and easy to interpret and can be automated. PMID- 9350763 TI - Isolation and characterization of a coronavirus from elk calves with diarrhea. AB - This is the first report of the isolation of a coronavirus from elk calves. Two fecal samples from elk calves with diarrhea were shown to be positive for coronavirus-like particles by electron microscopy, and the particles were propagated in the human rectal tumor-18 cell line. After 24 h, syncytia were observed, and cell culture supernatants from both samples showed hemagglutinating activity with mouse erythrocytes. Cells infected with both elk coronavirus (ECV) isolates reacted with Z3A5, a monoclonal antibody against the spike protein of bovine coronavirus (BCV), on an indirect fluorescent antibody test. The protein profiles of both ECV isolates were similar to that of BCV as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. On Northern blot analysis, the transcriptional pattern of ECV was typical of coronaviruses, with a nested set of transcripts with common 3' end sequences. Based on a published nucleoprotein gene sequence for BCV (Mebus isolate), we arbitrarily designed two primers for amplification by PCR. After cloning, the nucleoprotein was sequenced and a high degree of homology (99%) between the nucleoprotein gene sequences of ECV and BCV was observed. Thus, ECV is closely related genetically and antigenically to BCV and will be a new member of antigenic group 2 of the mammalian coronaviruses, which possess hemagglutinin-esterase protein. PMID- 9350761 TI - Rapid diagnosis of human brucellosis by peripheral-blood PCR assay. AB - A single-step PCR assay with genus-specific primers for the amplification of a 223-bp region of the sequence encoding a 31-kDa immunogenetic Brucella abortus protein (BCSP31) was used for the rapid diagnosis of human brucellosis. We examined peripheral blood from 47 patients, with a total of 50 cases of brucellosis, and a group of 60 control subjects, composed of patients with febrile syndromes of several etiologies other than brucellosis, asymptomatic subjects seropositive for Brucella antibodies, and healthy subjects. Diagnosis of brucellosis was established in 35 cases (70%) by isolation of Brucella in blood culture and in the other 15 cases (30%) by clinical and serological means. The sensitivity of our PCR assay was 100%, since it correctly identified all 50 cases of brucellosis, regardless of the duration of the disease, the positivity of the blood culture, or the presence of focal forms. The specificity of the test was 98.3%, and the only false-positive result was for a patient who had had brucellosis 2 months before and possibly had a self-limited relapse. In those patients who relapsed, the results of our PCR assay were positive for both the initial infection and the relapse, becoming negative once the relapse treatment was completed and remaining negative in the follow-up tests at 2, 4, and 6 months. In conclusion, these results suggest that the PCR assay is rapid and easy to perform and highly sensitive and specific, and it may therefore be considered a useful tool for diagnosis of human brucellosis. PMID- 9350764 TI - Detection and identification of Candida species in experimentally infected tissue and human blood by rRNA-specific fluorescent in situ hybridization. AB - Two 18S rRNA-targeted oligonucleotide probes specific for Candida albicans and Candida parapsilosis were used to detect and identify by fluorescent in situ hybridization these medically important Candida species in deep organs of mice after experimental systemic infection. The C. albicans-specific probe detected fungal cells in kidney, spleen, and brain sections of a mouse infected with C. albicans but not in a mouse infected with the closely related species C. parapsilosis. Conversely, the C. parapsilosis-specific probe detected fungal cells in the deep organs of a mouse infected with C. parapsilosis but not in the deep organs of a C. albicans-infected mouse. In addition, the C. albicans specific probe was used to detect this species in human blood spiked with yeast cells by a lysis-filtration assay and subsequent fluorescent in situ hybridization. By this assay, as few as three yeast cells per 0.5 ml of blood were consistently detected. Our results demonstrate that fluorescent in situ hybridization with species-specific rRNA-targeted oligonucleotide probes provides a novel, culture-independent method for the sensitive detection and identification of Candida species in clinically relevant material. PMID- 9350765 TI - Scytalidium dimidiatum and Lecythophora hoffmannii: unusual causes of fungal infections in a patient with AIDS. AB - Immunocompromised patients are susceptible to infections by fungi that seldom cause disease in humans. We describe a human immunodeficiency virus-infected patient who had simultaneous infections with two fungi which are rare causes of serious infection: Lecythophora hoffmannii, causing chronic sinusitis, and Scytalidium dimidiatum, causing skin lesions, lymphangitis, and lymphadenitis. The clinical and pathologic findings are discussed. PMID- 9350767 TI - Genes coding for enterotoxins and verotoxins in porcine Escherichia coli strains belonging to different O:K:H serotypes: relationship with toxic phenotypes. AB - Seventy-four E. coli strains isolated from piglets with diarrhea or edema disease in Spain were serotyped and examined for production of heat-labile (LT) and heat stable (ST) enterotoxins (LT-I, LT-II, STaH, STaP, and STb) and verotoxins (VT1, VT2, and VT2v = VTe) by phenotypic (Vero cell assay and infant mouse test) and genotypic (colony hybridization and PCR) methods. In general, an excellent correlation was found between the results obtained with a PCR approach and those determined with biological assays. DNA probes used in the hybridization also showed a very good agreement with phenotypic results, with the exception of a VT1 probe that initially produced 10 false-positive reactions. The gene coding for STb (58 strains) was the most prevalent gene detected by PCR, followed by those coding for STa (46 strains), LT (19 strains), VT2v (11 strains), and VT1 (1 strain). Apparently, in Spain three seropathotypes are predominant: (i) O149:K91:H10 K88+ LT-I+ STb+, (ii) O141:K85ab:H- P987+ STaP+, and (iii) O138:K81:H14 or H- STaP+ VT2v+. We conclude that PCR is a fast, specific, and practical method for the identification of enterotoxin and VT genes in clinical and epidemiological studies. PMID- 9350766 TI - Production of toxins (enterotoxins, verotoxins, and necrotoxins) and colicins by Escherichia coli strains isolated from septicemic and healthy chickens: relationship with in vivo pathogenicity. AB - Since the mechanism of virulence of Escherichia coli strains pathogenic to birds is not fully understood, the prevalence of toxic factors produced by E. coli strains pathogenic to other animals was investigated. A total of 625 E. coli strains isolated from visceral organs of chickens with colisepticemia and from feces of healthy chickens in Spain were tested for production of enterotoxins (heat labile [LT] and heat stable [STa]), verotoxins (VT1, VT2, and VT2v), cytotoxic necrotizing factors (CNF1 and CNF2), alpha-hemolysin (Hly), enterohemolysin (EntHly), colicin V (Col V) and other types of colicins, and necrotic and lethal activities. Only 45 (7%) of avian E. coli strains were toxigenic: 20 strains produced a cytotoxic response in HeLa but not in Vero cells, indicating the production of a cytotoxin not related to the VTs; 16 were EntHly+; 5 produced a new cytotonic product that causes the appearance of whitish vacuola in Vero and HeLa cells; 3 synthesized soluble factors that cause lethal activity in mice; and 1 elaborated LT. None of 625 avian E. coli strains was positive for production of VTs or CNFs. In contrast, colicinogenicity occurred in 335 (73%) of the 458 septicemic strains and 97 (58%) of 167 fecal isolates (P < 0.01), and this property was correlated with in vivo pathogenicity of strains. Thus, 80% (P < 0.001) and 66% (P < 0.001) of strains producing Col V and other types of colicins were characterized as being of high pathogenicity, whereas only 15% of the noncolicinogenic strains were classified as highly pathogenic. Our results clearly support the special pathogenicity theory, because 60% of the E. coli strains belonging to 18 serogroups (O1, O2, O5, O8, O12, O14, O15, O18, O20, O53, O78, O81, O83, O102, O103, O115, O116, and O132) most frequently identified among clinical septicemic strains were classified as highly pathogenic in in vivo assays, whereas only 24% of the strains with O serogroups less prevalent among diseased chickens were considered highly pathogenic (P < 0.01). PMID- 9350768 TI - Application of immunohistochemistry and in situ hybridization for detection of bovine coronavirus in paraffin-embedded, formalin-fixed intestines. AB - A monoclonal antibody (MAb) (Z3A5) against spike protein subunit of bovine coronavirus (BCV) reacted with the virus in formalin-fixed intestines in an immunoperoxidase test. We found an 88% correlation between immunohistochemistry with Z3A5 and in situ hybridization with a BCV nucleoprotein cDNA probe. MAb Z3A5 reacted with 90 BCV isolates from the United States and was an effective reagent for the diagnosis of BCV. PMID- 9350769 TI - Strains of glycopeptide-resistant Enterococcus faecium can alter their van genotypes during an outbreak. AB - Two isolates of Enterococcus faecium with VanA glycopeptide resistance were isolated during a hospital outbreak of E. faecium with plasmid-mediated VanB resistance. Both were found to be identical to the VanB outbreak strain by pulsed field gel electrophoresis. The genotype of this strain changed from vanB to vanA through an intermediate isolate that contained both the vanA and vanB gene clusters on distinct plasmids. PMID- 9350770 TI - Rapid identification of mycobacteria to species level by PCR-restriction fragment length polymorphism analysis of the hsp65 gene and proposition of an algorithm to differentiate 34 mycobacterial species. AB - PCR-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene (A. Telenti, F. Marchesi, M. Balz, F. Bally, E. C. Bottger, and T. Bodmer, J. Clin. Microbiol. 31:175-178, 1993) was applied to 108 mycobacterial isolates representing 34 species to evaluate its potential as a rapid reference method. A total of 49 distinct patterns were obtained; 25 species were characterized by a single PRA pattern, while 9 species gave more than one specific pattern. An algorithm describing these 34 species (which includes five additional species and new subgroups of Mycobacterium kansasii, M. abscessus, and M. peregrinum) is proposed. A relatively simple and inexpensive method, PRA may be particularly helpful in routine clinical microbiology laboratories. PMID- 9350771 TI - The aerobic and anaerobic bacteriology of perirectal abscesses. AB - The microbiology of perirectal abscesses in 144 patients was studied. Aerobic or facultative bacteria only were isolated in 13 (9%) instances, anaerobic bacteria only were isolated in 27 (19%) instances, and mixed aerobic and anaerobic flora were isolated in 104 (72%) instances. A total of 325 anaerobic and 131 aerobic or facultative isolates were recovered (2.2 anaerobic isolates and 0.9 aerobic isolates per specimen). The predominant anaerobes were as follows: Bacteroides fragilis group (85 isolates), Peptostreptococcus spp. (72 isolates), Prevotella spp. (71 isolates), Fusobacterium spp. (21 isolates), Porphyromonas spp. (20 isolates), and Clostridium spp. (15 isolates). The predominant aerobic and facultative bacteria were as follows: Staphylococcus aureus (34 isolates), Streptococcus spp. (28 isolates), and Escherichia coli (19 isolates). These data illustrate the polymicrobial aerobic and anaerobic microbiology of perirectal abscesses. PMID- 9350772 TI - Rapid pulsed-field gel electrophoresis protocol for typing of Escherichia coli O157:H7 and other gram-negative organisms in 1 day. AB - Genomic DNA patterns generated by pulsed-field gel electrophoresis are highly specific for different strains of an organism and have significant value in epidemiologic investigations of infectious-disease outbreaks. Unfortunately, time consuming and tedious specimen processing is an inherent problem which limits the use of this powerful technology as a real-time epidemic investigational tool. Here, I describe a rapid method to improve the response time and provide specific bacterial strain identification for the typing of Escherichia coli O157:H7 and other gram-negative organisms in a single day. PMID- 9350773 TI - Virulence patterns of Escherichia coli K1 strains associated with neonatal meningitis. AB - The prevalence of the ibe10 gene, of the pap, afa, and sfa adhesin-encoding operons, and of a 14.9-kb rrn-containing HindIII fragment was studied for 67 Escherichia coli neonatal meningitis strains, 58 E. coli K1 commensal strains, and 47 E. coli blood isolates from neonates without meningitis. ibe10, sfa, and the 14.9-kb HindIII fragment were observed significantly more often in the meningitis strains than in blood or commensal strains. PMID- 9350774 TI - Evaluation of the automated COBAS AMPLICOR hepatitis C virus PCR system. AB - To evaluate the reliability and feasibility of the automated Roche COBAS AMPLICOR PCR system for routine detection of hepatitis C virus (HCV) RNA, a total of 405 serum samples previously tested by an in-house nested PCR and manual Roche AMPLICOR microwell plate HCV test were examined. Complete concordance was found between the results with the HCV COBAS AMPLICOR system and the previously determined HCV RNA status. The automated HCV COBAS AMPLICOR system provides the clinical microbiology laboratory with a specific and sensitive PCR method for rapid and reliable detection of HCV RNA. PMID- 9350776 TI - Surveillance and detection of erythromycin resistance in Bordetella pertussis isolates recovered from a pediatric population in the Intermountain West region of the United States. AB - Forty-seven Bordetella pertussis isolates recovered from January 1985 to June 1997 at Primary Children's Medical Center were tested for erythromycin resistance. Agar dilution MICs were determined on Regan-Lowe agar. Forty-six isolates were found to be erythromycin susceptible (all MICs were less than or equal to 0.12 microg/ml). One isolate was found to be erythromycin resistant (MIC, 32 microg/ml). In addition, we compared Etest MIC results and disk diffusion zone diameter measurements, performed on commercially prepared Regan Lowe agar, to the agar dilution MIC result. Etest MIC and/or disk diffusion testing on commercial Regan-Lowe agar appears to be an adequate method for erythromycin resistance screening of B. pertussis isolates. PMID- 9350775 TI - Development of an antimicrobial susceptibility surveillance system for Neisseria gonorrhoeae in Malawi: comparison of methods. AB - Susceptibility of Neisseria gonorrhoeae to gentamicin, the primary treatment for gonorrhea in Malawi since 1993, was determined by using agar dilution MICs, E test MICs, disc diffusion, and clinical cure rate. Agar dilution MICs were slightly higher in 1996 than in 1993 isolates, with a concomitant drop in the clinical cure rate. E-test MICs were substantially lower than agar dilution determinations, with only 77.4% within 1 log2 concentration. PMID- 9350777 TI - Tilletiopsis minor: a new etiologic agent of human subcutaneous mycosis in an immunocompromised host. AB - We describe herein the isolation of Tilletiopsis minor from a subcutaneous cyst of a 70-year-old immunocompromised male. The diagnosis was based on repeated isolation of the fungus, observation of hyphal elements in tissue sections, the ability of the mold to grow at or near body temperature, and the achievement of a complete cure following surgery and antifungal therapy. PMID- 9350778 TI - Serotyping of group A rotaviruses in Egyptian neonates and infants less than 1 year old with acute diarrhea. AB - Group A human rotavirus G serotypes were detected in stool specimens from neonates and infants with and without acute diarrhea in Cairo by using monoclonal antibodies in an enzyme-linked immunosorbent assay. Serotypes G1 and G4 predominated in all age groups. Mixed (G1 plus G4) and nontypeable specimens represented 16.1 and 38.7% of the total number serotyped, respectively. PMID- 9350779 TI - Immunoglobulin A antibodies to Helicobacter pylori. AB - Serological testing for immunoglobulin G (IgG) antibodies to Helicobacter pylori has proven useful in supporting the diagnosis of infection with this organism, but the clinical value of IgA antibodies in H. pylori-related gastritis remains controversial. The purpose of our study was to determine the frequency of IgA positive IgG-negative patients with symptoms of gastrointestinal (GI) disorders, thus assessing the clinical utility of IgA testing for H. pylori-related gastritis. It was found previously that the frequency of infected individuals in this category (IgA positive and IgG negative) is about 2%, but a large number of IgG-negative patients with GI disorders suggestive of H. pylori infection have not been investigated until now. PMID- 9350780 TI - Characterization to species level of Mycobacterium avium complex strains from human immunodeficiency virus-positive and -negative patients. AB - Forty human clinical Mycobacterium avium-M. intracellulare complex strains isolated in Greece were characterized to the species level by PCR with three sets of primers specific for one or both species. M. avium predominated in both human immunodeficiency virus-positive and -negative patients, but the frequency of M. intracellulare isolation appeared to be higher in the latter. PMID- 9350782 TI - Proline-aminopeptidase test for rapid screening of Clostridium difficile. PMID- 9350781 TI - Comparison of charcoal- and starch-based media for testing susceptibilities of Legionella species to macrolides, azalides, and fluoroquinolones. AB - We compared growth characteristics of 46 Legionella strains grown on buffered charcoal yeast extract alpha (BCYE alpha) agar and buffered starch yeast extract (BSYE) agar and MICs of macrolides, azalides, and fluoroquinolones for these organisms. Growth was poor and not reproducible on BSYE agar. Growth was excellent on BCYE alpha, and MICs were easy to interpret. BCYE alpha is superior to BSYE for testing susceptibilities of Legionella species by agar dilution. PMID- 9350783 TI - Amplification-based DNA fingerprinting: from artifactual to definitive typing and in between. PMID- 9350785 TI - Candida dubliniensis. PMID- 9350784 TI - Absence of human herpesvirus 8 DNA in benign and malignant endothelial lesions. PMID- 9350786 TI - Research and clinical issues in chronic venous disease. AB - The purpose of this study was to summarize the current issues in chronic venous disease by reviewing of the literature relating to the condition. The review was conducted by members of the Committee on Research of the American Venous Forum and includes the Committee's venous disease and current/future directions. Progress in the understanding and management of chronic venous problems has lagged behind that in arterial disease, despite the large number of patients affected. The complex pathophysiology of venous problems, lack of accepted evaluation standards and lack of prosthetic conduits are some of the factors which contribute to this. New information in these areas has laid the foundation for advances in both operations and non-operative therapy. In conclusion, many opportunities for clinical and basic research in the area of chronic venous disease are available. Application of basic science techniques, including those of molecular biology, will lead to new insights into pathophysiology of chronic venous syndrome. Developments in technology, classification and basic science suggest multiple new potentials therapeutic approaches in the next decade. PMID- 9350788 TI - Celebration of the 50th anniversary of endarterectomy: the operation of Joao Cid dos Santos. PMID- 9350787 TI - Dipyridamole: an unfulfilled promise? PMID- 9350789 TI - Correlating clinical indicators of lower-limb ischaemia with quality of life. AB - The objectives of the study were to analyse the impact of increasing lower-limb ischaemia upon quality of life and to assess the correlation between clinical indicators of lower-limb ischaemia and such quality. A prospective observational study of a consecutive series of 235 patients (144 men and 91 women; median age 68 (range 41-87) years presenting with varying degrees of lower-limb ischaemia graded according to ISCVS criteria was performed. Data was collected at interview before any intervention. Clinical indicators of lower-limb perfusion included: intermittent claudication and maximum walking distance on standardized treadmill testing; ankle:brachial pressure indices and isotope limb blood flow. Quality of life analysis was performed using the EuroQol (EQ) questionnaire. This is a standardized generic instrument for describing health-related quality of life and consists of a descriptive system of five dimensions, each measured on three levels. Thus, a profile and two single indices of quality of life were derived using different methods. Increasing lower-limb ischaemia results in a statistically significant deterioration in both global quality of life and in all EQ-measured quality of life dimensions (P < 0.01 Kruskal-Wallis, ANOVA). The correlation between clinical indicators and quality of life is statistically significant but not sufficiently close (correlation coefficients < 0.6) to assume that variations in clinical indicators result in reciprocal variations in quality of life. In conclusion, as might be expected, a significant correlation exists between clinical indicators of lower-limb ischaemia and health-related quality of life. However, the low correlation coefficients emphasize how tenuous the association is. Thus, a significant improvement in the clinical indicators of lower-limb ischaemia cannot be assumed to impart a similar benefit on quality of life. The latter concept must therefore be analysed independently. PMID- 9350790 TI - Internal mammary artery graft function is not affected in hypertensive patients on therapy. AB - Results are presented which assess the reactivity of isolated human internal mammary artery fragments from non-hypertensive and treated hypertensive patients in vitro. Material from three patient groups was examined: group I, no hypertension; group II, arterial hypertension treated with ACE inhibitors; and group III, arterial hypertension treated with nifedipine. Responses to KCl, norepinephrine and acetylcholine, as well as the influence of N(G)-monomethyl-L arginine (L-NMMA) on the effects of norepinephrine were tested. Response to KCl was highest in group III, while the contractile reactivity to norepinephrine was depressed in group II. Relaxation after acetylcholine was enhanced in groups II and III. Incubation of vessel fragments with L-NMMA sensitized the tissue to norepinephrine in the order of potency group II>group III>group I. Internal mammary artery function as the graft, and particularly in terms of endothelial function, is not adversely affected in arterial hypertension, although proper antihypertensive treatment may be essential. PMID- 9350791 TI - Prediction of requirement for, and outcome of, prolonged mechanical ventilation following cardiac surgery. AB - The prediction of requirement for, and short- and long-term outcome of, prolonged mechanical ventilation after cardiac surgery is ill-defined. The aims of this study were to isolate any predictive indices which might identify those groups of patients who may require prolonged ventilation postoperatively and to determine which factors significantly affect outcome in the prolonged-ventilation group. Following case note review of 139 consecutive cardiac surgical patients ventilated for > or = 7 days following surgery, 43 factors were recorded on each patient, including smoking, pulmonary function, chest infection, and chronic obstructive airways disease. Of 139 patients, 89 were discharged from hospital (64% survival); of these, 52 were alive at long-term follow-up (58% long-term survival). Statistical analysis identified urban residence, chronic obstructive airways disease, prolonged operation, and bypass time as significant predictors of requirement for prolonged ventilation postoperatively. On multivariate analysis five factors were predictive of increased intensive care mortality, including urban residence, inotrope days, sepsis, perioperative cerebrovascular accident and coagulopathy requiring fresh frozen plasma transfusion postoperatively. Following discharge from hospital, four factors were found to be significant predictors of increased mortality: these are impaired preoperative ejection fraction, increasing age, impaired preoperative pulmonary function, and abscence of preoperative aspirin medication. These factors should be considered in intensive care planning, long-term follow-up and importantly on clinical decision making in the individual patient. PMID- 9350792 TI - Myocardial revascularization without cardiopulmonary bypass in pigs: a comparison between beta-blockers and Ca2+-channel blockers. AB - The purpose of this study was to develop the surgical technique for internal mammary artery grafting on the beating heart. In 10 pigs, heart rate was reduced with esmolol (n = 5) or verapamil (n = 5). In addition, the anti-ischaemic and anti-arrhythmic potencies of these drugs were investigated. Haemodynamics, mechanical function and ischaemia-sensitive laboratory measurements were assessed perioperatively. There were no differences in pre-ischaemic data including ischaemic clamping time of the left anterior descending artery. At the end of ischaemia haemodynamic parameters were significantly lower in the esmolol-group (P < 0.05). In the verapamil-group, one pig died from ventricular fibrillation during ischaemia, and one showed fibrillation during reperfusion (P < 0.01). There were no differences in cardiac function or enzymes between the groups. Reduction of heart rate was provided by both drugs, but no conclusive evidence was provided with regard to additional anti-ischaemic and anti-arrhythmic protection. PMID- 9350793 TI - Bifurcated stent-graft for abdominal aortic aneurysm. AB - The purpose of this study was to examine bifurcated stent-graft implantation for abdominal aortic aneurysm. Fifty-seven patients were treated and followed with serial computed tomography scans for up to 3 years. Patients were allocated to three groups (first 20, second 20, last 17) according to when the repair was performed. Successful treatment is defined as exclusion of the aneurysm from the circulation, based on contrast computed tomography. Success rates in the three groups were 55%, 70% and 100%. Perigraft leak (endoleak) was present on initial assessment in 4/20, 2/20 and 1/17. Two of these aneurysms ruptured early in the postoperative period. Thereafter, leaks were sought and treated aggressively. Kinking and thrombosis occurred in six of the first 20 patients, but did not occur in any of the last 37 patients, in whom the graft limbs were routinely stent-supported throughout. Infrarenal implantation in very a short neck (< 10 mm), or a thrombus-lined neck was associated with proximal stent migration. In conclusion, changes in patient selection and technique have led to a steady rise in the short-term success rate of the stent-grafted implantation. PMID- 9350794 TI - Stent grafts for iliofemoral occlusive disease. AB - This report summarizes the technical feasibility and early results of endovascular iliofemoral stented grafts in the treatment of iliofemoral occlusive disease. Twenty-four patients (mean age 71 years) underwent 29 lower-extremity inflow procedures for claudication (n = 7) or limb threatening ischaemia (n = 17). The technical success rate for endovascular grafts was 93% (n = 27). Some 85% of the grafts originated from the aortoiliacjunction or the common iliac arteries. Outflow procedures were performed in all cases and consisted of profundaplasty (n = 17) and/or femorodistal grafting (n = 13). The operative mortality rate was 9% and one occlusion was noted in the early postoperative period. The mean (s.d.) primary and secondary cumulative patency rates after 1 year were 85(10)% and 95(5)% respectively. The corresponding limb salvage rate was 95(4)%. The authors conclude that endovascular iliofemoral stented grafts through a single groin incision are technically feasible and that early patency rates are acceptable. More experience is needed however before widespread application of these new techniques can be justified. PMID- 9350796 TI - The thrombosed arteriovenous graft: an endovascular model for vascular surgeons. AB - Arteriovenous dialysis grafts are the most commonly implanted prosthetic grafts. Thrombectomy with selective graft revision is traditional therapy for occlusions, but patency is minimally prolonged. Stenoses are determined by tactile feedback from an embolectomy catheter and lack of prograde and retrograde bleeding. An objective method for studying the graft and inflow and outflow tracts that permits appropriate endoluminal or surgical correction is described. This approach is appealing because: (i) the current approach is inadequate; (ii) it offers an objective, quantitative method to determine frequency and severity of critical stenoses within the failed access graft; (iii) remote and perigraft stenoses can be treated at the same setting; and (iv) it promotes the development of endovascular skills by surgeons in a high-volume, low-risk setting. PMID- 9350795 TI - Closed superficial femoral artery endarterectomy: a 2-year follow-up. AB - Closed superficial femoral artery endarterectomy is a minimal invasive technique. Through a single groin incision the occluded intima is remotely removed by a newly modified ring stripper. This device has a double ring which acts like remote scissors and enables the surgeon to recanalize long segmental femoral popliteal occlusive disease. The device was used in 103 limbs in 90 patients. The cumulative assisted primary and secondary patency was 86%. This technique is an alternative to bypass grafting when vein is not available. PMID- 9350797 TI - Utility of carotid duplex in young adults with lower extremity atherosclerosis: how aggressive should we be in screening young patients? AB - The purpose of this study was to determine the prevalence and degree of carotid disease in patients with premature lower-extremity atherosclerosis. Seventy-six young men (mean age at onset of symptoms 42+/-0.5 years with premature lower extremity atherosclerosis who underwent complete carotid duplex scans were studied. The mean lowest ankle: brachial index was 0.49+/-0.02. Forty-seven patients (62%) required interventions to treat advanced leg symptoms, and 18 (24%) experienced disease progression during the study period. Carotid duplex scans showed internal carotid occlusions in eight (11%); advanced or critical plaque disease (60-99% diameter loss) in 14 (18%); moderate plaque disease (40 59% diameter loss) in 16 (21%); mild plaque disease (intimal thickening or 1-39% diameter loss) in 18 (24%); and normal carotid arteries in 20 (26%). Comparing the 20 subjects with normal carotid arteries to the S6 with any evidence of disease, there were no differences in age of onset, risk factors, coronary artery disease, mean ankle: brachial index, number of interventions, disease progression, amputation, or death. Fifteen (27%) of the patients with carotid atherosclerosis ultimately developed transient ischemic attack or stroke; 13 of these had advanced carotid stenoses or carotid occlusions. In conclusion, carotid plaque disease is prevalent among patients with premature atherosclerosis of the lower extremity. The presence of carotid atherosclerosis is not related to the degree of lower extremity atherosclerosis, nor to the rate of disease progression. Carotid duplex scans are indicated to screen these young patients for compelling lesions that might warrant prophylactic carotid endarterectomy. PMID- 9350798 TI - A critical approach for longitudinal clinical trial of stretch PTFE aortic grafts. AB - Adaptation of new clinical products should be based upon thorough scientific evaluation of properties and performance in vitro and in vivo. Developmental animal research experimentation is classically carried out by the manufacturer with eventual government approval. However, objective data needs to be recorded during clinical trials including handling characteristics, bleeding, tensile strength, kinking, seamline break, dilatation, anastomotic deterioration, patency, and incorporation. Since April 1991, 1010 stretch polytetrafluoroethylene (PTFE) aortic grafts have been implanted at our institution and data were recorded prospectively. Six hundred and seven were for elective abdominal aortic aneurysms, 46 for symptomatic abdominal aortic aneurysms, 58 for ruptured abdominal aortic aneurysms, 17 for elective thoracoabdominal aneurysms, 3 for ruptured thoracoabdominal aneurysms and the remainder were for various aortoiliac pathology. Average age of the patients was 69 (range: 10-95), 66% were males, 25% were diabetics. Overall operative mortality was 5.8% (2.9% in elective cases and 26.6% in emergent cases). There were 23 occlusions; 21 were revised and 2 were replaced with axillofemoral bypasses. Estimated blood loss was 784 cc in elective cases and 1918 cc in emergent cases. Grafts were followed by duplex ultrasound or CT scan every 3 months during the first year and every 6 months thereafter. There were no graft dilatations or false aneurysms in this series. There was one graft infection and no perigraft seromas or anastomotic deteriorations during this follow up. Follow up was complete in 94% of these patients. In conclusion, stretch PTFE graft has acceptable handling characteristics, no excessive bleeding at the suture line and had no anastomotic or graft dilatation. This graft material was suitable for thoracic, visceral, renal and abdominal aortic reconstructions. PMID- 9350799 TI - Re-do operations after failed multisegmental reconstructive arterial surgery for critical limb ischaemia. AB - The purpose of this study was to investigate the long-term graft patency rates after multisegmental arterial reconstruction for treatment of chronic critical limb ischemia, and to evaluate the role of re-do surgery in treatment of graft failure. A total of 449 aortofemoropopliteal/tibial grafts carried out over a 10 year period were retrospectively reviewed. All patients were operated upon with chronic critical limb ischemia grade III and IV according to the Fontaine classification; 221 operations were performed in one stage (group A), and 228 in two stages (group B). Distribution of graft failures in the postoperative period, re-do operations and their impact on limb salvage were investigated using life table methods. During follow up, 62 cases of inflow graft thrombosis were observed (23 in group A and 39 in group B). To correct the inflow graft failure, 59 re-do procedures were performed (27 in group A, 32 in group B). Inflow graft failures were most common during 24 months after primary surgery. During the same period, 92 cases of isolated outflow graft thrombosis were observed (45 in group A and 47 in group B). Outflow graft thromboses were most common after 24-36 months. For treatment of recurrent symptoms caused by outflow graft thrombosis, 68 re-do operations were performed. The 5 year cumulative primary graft patency, secondary graft patency and limb salvage rates were 43.2%, 71.8% and 79.9% in group A, and 23.8%, 54% and 67.5% in group B respectively. In conclusion the long term primary graft patency rate after multisegmental aortofemoropopliteal/tibial reconstructive surgery is low and significantly lower, when compared with single segment reconstructions. Re-do operations have a positive impact on secondary long-term graft patency and limb salvage. PMID- 9350800 TI - Quality of life changes after angioplasty for claudication: medium-term results affected by comorbid conditions. AB - Rapid improvements in walking distance and quality of life have been identified in patients with intermittent claudication following percutaneous transluminal angioplasty, but the medium-term results are less well defined. The aim of this study was to assess quality of life and walking distance in the medium term. Walking distance was assessed before percutaneous transluminal angioplasty and at 6 weeks and 1 year after the procedure using a previously validated questionnaire. At the same time, quality of life was assessed using a EuroQol questionnaire and a visual analogue scoring system. Twenty-four patients (12 men, 12 women, mean age 65 years) underwent successful percutaneous transluminal angioplasty (five iliac, 17 femoropopliteal, two both). Significant improvements in walking distance and quality of life were demonstrated following percutaneous transluminal angioplasty. These were maintained at 1 year, although perceived health state deteriorated. During the study period, six patients developed other serious comorbidities. Development of comorbid conditions may affect the medium term outcome of quality of life studies in patients treated for intermittent claudication. Data from such studies should therefore be interpreted with care. PMID- 9350801 TI - Annular dilatation increases stress in the mitral valve and delays coaptation: a finite element computer model. AB - The purpose of this study was to examine the effects of annular dilatation on coaptation, and leaflet and chordal stresses, using a three dimensional finite element computer model. To do this, the whole mitral valve was simulated using ANSYS 4.4A software. Normal model geometry, collagen fiber orientation, tissue thickness, and material properties were determined from fresh porcine valves. For annular dilatation, the annular circumference was increased by 18% versus normal. Isovolumic contraction and rapid ventricular ejection were simulated. Data showed that, in the annular dilatation model, the stress magnitudes increased more than two-fold compared with normal in both the anterior leaflet and posterior leaflet. Coaptation was greatly delayed in the dilatation model, and the leaflets never fully coapted. Chordal stresses were also greatly increased in the dilatation model. In conclusion, increased stress due to annular dilatation may lead to tissue disruption, further dilatation, delayed coaptation, and increased regurgitation, in a 'closed-loop' degenerative process. PMID- 9350802 TI - Differential patterns of atherosclerotic disease in patients with unilateral hemiparesis resulting from poliomyelitis: case reports demonstrating the possible effect of hemodynamics. AB - The relative importance of hereditary and mechanical factors in the pathogenesis of aneurysms remains as controversial today as it was two decades ago. The cases of two patients with unilateral paresis resulting from poliomyelitis who presented with abdominal aortic aneurysms are reported. In addition, each patient had iliofemoral aneurysms contralateral to, and iliofemoral occlusive disease ipsilateral to, their affected extremity. The two cases detailed within this report suggest that hemodynamic forces may alter the pattern of disease in arteries affected by arteriosclerosis. PMID- 9350803 TI - Aortico-left ventricular tunnel: late reoperations. AB - Aortico-left ventricular tunnel is a rare congenital anomaly that presents as aortic regurgitation in infancy or childhood. Information on late results of surgery, especially with reoperations for progressive aortic regurgitation and recurrences are limited. The case histories of two patients with reoperations following correction of aortico-left ventricular tunnel are reported. PMID- 9350804 TI - Basic fibroblast growth factor promotes extension of regenerating axons of peripheral nerve. In vivo experiments using a Schwann cell basal lamina tube model. AB - Schwann cell basal lamina tubes serve as attractive conduits for regeneration of peripheral nerve axons. In the present study, by using basal lamina tubes prepared by in situ freeze-treatment of rat saphenous nerve, the effects of exogenously applied basic fibroblast growth factor (bFGF) on peripheral nerve regeneration was examined 2 and 5 days after bFGF administration. Regenerating axons were observed by light and electron microscopy using PGP9.5 immunohistochemistry for specific staining of axons. In addition, the localizations of bFGF and its receptor (FGF receptor-1) were examined by immunohistochemistry using anti-bFGF antibody and anti-FGF receptor-1 antibody, respectively. Regenerating axons extended further in the bFGF-administered segment than in the bFGF-untreated control segment. Electron microscopy showed that regenerating axons grew out unaccompanied by Schwann cells. Findings concerning angiogenesis and Schwann cell migration were very similar between the bFGF treated and control nerve segment. bFGF-immunoreactivity was not detected in the control nerve segment. In contrast, bFGF-immunoreactivity was detected on the basal lamina tubes as well as on the plasmalemma of regenerating axons facing the basal lamina in the bFGF treated nerve segment up to 5 days after administration, suggesting that exogenous bFGF can be retained in the basal lamina for several days after administration. FGF receptor was detected on the plasma membrane of regenerating axons where they abutted the basal lamina. These results indicate that bFGF could promote the extension of early regenerating axons by directly influencing the axons, but not via Schwann cells or angiogenesis. PMID- 9350805 TI - Organization of the cortical endoplasmic reticulum in the squid giant axon. AB - The organization of the cortical endoplasmic reticulum in the squid giant axon was investigated by rapid freeze and freeze-substitution electron microscopy, thereby eliminating the effects of fixatives on this potentially labile structure. Juvenile squid, which have thinner Schwann sheaths, were used in order to achieve freezing deep enough to include the entire axonal cortex. The smooth endoplasmic reticulum is composed of subaxolemmal and deeper cisternae, tubules, tethers and vesicles. The subaxolemmal cisternae make junctional contacts with the axolemma which are characterized by filamentous-granular bridging structures approximately 3 nm in diameter. The subaxolemmal junctions with the axolemma resemble the coupling junctions between the sarcoplasmic reticulum and the T tubules in muscle. Reconstruction of short series of sections showed that a number of the elements of the endoplasmic reticulum were continuous but numerous separate vesicles were present as well. The morphology of endoplasmic reticulum as described here suggests that it is a highly dynamic entity as well as a Ca2+ sequestering organelle. PMID- 9350806 TI - Loss and survival of spiral ganglion neurons in the guinea pig after intracochlear perfusion with aminoglycosides. AB - Loss of cochlear hair cells results in a loss of ganglion cells and further neurodegenerative changes throughout the auditory pathway. Understanding more about the early stages of ganglion cell loss in vivo may lead to ways of ameliorating or preventing the loss of these neurons. To examine these stages, the effects of intracochlear perfusion with aminoglycoside antibiotics on the organ of Corti and spiral ganglion cells were evaluated in young adult guinea pigs at survival periods ranging from 1 hour to 12 weeks, using immunocytochemical and ultrastructural techniques. At 1 hour survival a base-to apex gradient of damage was indicated in the cochlea by the appearance of severely damaged hair cells and injured ganglion cells in the basal coil while in the apical coil, hair cells were damaged but intact and ganglion cells appeared normal. By 4 hours the appearance of severely disrupted hair cells and damaged ganglion cells had extended throughout the cochlea. The ultrastructural appearance of many injured ganglion cells demonstrated features characteristic of cell death including condensed cytoplasm, non-marginal clumping of nuclear chromatin, and wrinkled nuclear membrane. Despite the loss of many ganglion cells, a population of these cells remained at 12 weeks survival. These contained large amounts of rough endoplasmic reticulum, were unmyelinated apart from the central process and were surrounded by satellite cells. These features are typical of ganglion cells during development, before the onset of hearing. Immunolabelling of cochlear whole mounts after hair cell destruction with protein gene product 9.5 (PGP 9.5) revealed the presence of neural elements in the organ of Corti at up to 12 weeks survival. These may be associated with the remaining ganglion cells. In these surviving ganglion cells, the intense labelling with PGP 9.5 together with the increase in rough endoplasmic reticulum, indicates the presence of active protein synthesis which may be connected with their survival. PMID- 9350807 TI - Hindshaker, a novel myelin mutant showing hypomyelination preferentially affecting the spinal cord. AB - Animals with spontaneous mutations affecting myelin formation have provided useful information about the genetic and cellular mechanisms regulating normal and abnormal myelination. In this paper we describe a novel murine mutation termed hindshaker (hsh), which is inherited in an autosomal recessive manner. Affected mice are characterised by a variable tremor of the hind end which commences at about 2 weeks of age and largely disappears in animals older than 6 weeks. There is hypomyelination affecting predominantly the spinal cord, although the optic nerves and brain are involved to a much lesser degree. The defect of thinly myelinated and naked axons is maximal at 20 days of age and largely resolves with time so that in the adult most axons are myelinated. The myelin structure appears normal and immunostains for the major proteins. Although the distribution of oligodendrocytes in the spinal cord is similar to normal during the period of hypomyelination, there are fewer mature cells. The hsh mutation appears to delay the maturation of oligodendrocytes, particularly in the spinal cord. Additionally, there is a considerable variation in phenotypic expression and in penetrance when the mutation is expressed on different genetic backgrounds, suggesting the hsh locus is subject to the influence of modifying gene(s). Identification of the hsh gene should identify a factor important in the development of oligodendrocytes, particularly those in the spinal cord. PMID- 9350808 TI - Development of endothelial paracellular clefts and their tight junctions in the pial microvessels of the rat. AB - The microvessels of the pia mater lack an investment with astrocyte processes but nonetheless have a high transendothelial electrical resistance which has caused them to be regarded as part of the blood-brain barrier. This high resistance is known to be acquired in the perinatal period. The aim of our study was to relate the known physiological changes with differentiation of the endothelial paracellular clefts and especially of their tight junctions which provide the basis for the high transendothelial resistance of blood-brain barrier vessels. Tight junctions of endothelial cell paracellular clefts in pial microvessels were examined by transmission electron microscopy using goniometric tilting to reveal and measure membrane separations at tight junctions in fetal, postnatal and adult rats. These tight junctional membrane separations narrowed over the period (E16: 6.3 nm, D1: 6.4 nm, D7: 5.4 nm) and differentiated into two groups by the adult stage: one with a membrane separation of 2.8 nm and the staining characteristics of non-brain endothelial junctions, and the other with no detectable membrane separation and the staining characteristics of blood-brain barrier endothelial junctions. This patchy and incomplete differentiation of pial tight junctions into a blood-brain barrier-like form could result either from non-uniform exposure to inductive signals or to local variation in responsiveness to such agents. Although these changes in junction organization may be related to the known increase in pial transendothelial resistance in the perinatal period, we have not yet identified any sharply defined structural change which coincides with this physiological event. PMID- 9350809 TI - Severe manifestations in carrier females in X linked retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a group of progressive hereditary disorders of the retina in which various modes of inheritance have been described. Here, we report on X linked RP in nine families with constant and severe expression in carrier females. In our series, however, the phenotype was milder and delayed in carrier females compared to hemizygous males. This form of X linked RP could be regarded therefore as partially dominant. The disease gene maps to chromosome Xp2.1 in the genetic interval encompassing the RP3 locus (Zmax=13.71 at the DXS1100 locus). Single strand conformation polymorphism and direct sequence analysis of the retinitis pigmentosa GTPase regulator (RPGR) gene, which accounts for RP3, failed to detect any mutation in our families. Future advances in the identification of X linked RP genes will hopefully help to elucidate the molecular basis of this X linked dominant RP. PMID- 9350810 TI - Spectrum of clinical features associated with interstitial chromosome 22q11 deletions: a European collaborative study. AB - We present clinical data on 558 patients with deletions within the DiGeorge syndrome critical region of chromosome 22q11. Twenty-eight percent of the cases where parents had been tested had inherited deletions, with a marked excess of maternally inherited deletions (maternal 61, paternal 18). Eight percent of the patients had died, over half of these within a month of birth and the majority within 6 months. All but one of the deaths were the result of congenital heart disease. Clinically significant immunological problems were very uncommon. Nine percent of patients had cleft palate and 32% had velopharyngeal insufficiency, 60% of patients were hypocalcaemic, 75% of patients had cardiac problems, and 36% of patients who had abdominal ultrasound had a renal abnormality. Sixty-two percent of surviving patients were developmentally normal or had only mild learning problems. The majority of patients were constitutionally small, with 36% of patients below the 3rd centile for either height or weight parameters. PMID- 9350812 TI - A syndrome including thumb malformations, microcephaly, short stature, and hypogonadism. AB - We report on eight patients from seven different families affected with a syndrome which includes thumb defects, short stature, microcephaly, and mental retardation. Most of the patients had additional malformations, in particular amenorrhoea and azoospermia in the adults. There were no haematological manifestations and the chromosomes were normal without evidence of breakage even after stimulation. In five of the cases the probands' mother received hormonal treatment before or at the beginning of her pregnancy or both. The syndrome may be inherited as an autosomal recessive trait since the patients included both males and females and their parents were related in most cases. In addition, supporting this possibility, they all originated from a small village which may be considered as an isolate. However, in all cases but one, only one person was affected in each family and there was a significant apparent excess of healthy sibs of the probands. These observations may be the result of the variability of the syndrome or a more complex type of inheritance. PMID- 9350813 TI - Characterisation of a satellited non-fluorescent Y chromosome (Y[nfqs]) by FISH. AB - A fetus was prenatally diagnosed as having a Y(nfqs) chromosome which was inherited from the father. With the QFQ technique, the Yqh was observed to be nonfluorescent and contained cytological satellites which were attached to the terminal long arm. The satellites were positively stained by the Ag-NOR technique suggesting that the NORs were active. A battery of DNA probes was used to characterise the Y(nfqs). Hybridisation experiments using a chromosome 15 specific classical satellite DNA probe (D15Z1) and a Yq telomere DNA probe showed that the additional satellited material on Yq originated from 15p, and that the Yq terminal region had been lost. This is the first reported case in which the origin of cytological satellites on Yq has been determined by FISH, but this does not imply that all satellited Y chromosomes are derived from 15p. However, the clinical significance of this Y(nfqs) chromosome remains obscure. PMID- 9350811 TI - The clinical and molecular genetic approach to Duchenne and Becker muscular dystrophy: an updated protocol. AB - Detection of large rearrangements in the dystrophin gene in Duchenne and Becker muscular dystrophy is possible in about 65-70% of patients by Southern blotting or multiplex PCR. Subsequently, carrier detection is possible by assessing the intensity of relevant bands, but preferably by a non-quantitative test method. Detection of microlesions in Duchenne and Becker muscular dystrophy is currently under way. Single strand conformational analysis, heteroduplex analysis, and the protein truncation test are mostly used for this purpose. In this paper we review the available methods for detection of large and small mutations in patients and in carriers and propose a systematic approach for genetic analysis and genetic counselling of DMD and BMD families, including prenatal and preimplantation diagnosis. PMID- 9350815 TI - Genetic heterogeneity of autosomal dominant polycystic kidney disease in Argentina. AB - Autosomal dominant polycystic kidney disease (ADPKD) is an inherited disorder with genetic heterogeneity. Up to three loci are involved in this disease, PKD1 on chromosome 16p13.3, PKD2 on 4q21, and a third locus of unknown location. Here we report the existence of locus heterogeneity for this disease in the Argentinian population by performing linkage analysis on 12 families of Caucasian origin. Eleven families showed linkage to PKD 1 and one family showed linkage to PKD2. Two recombinants in the latter family placed the locus PKD2 proximal to D4S1563, in agreement with data recently published on the cloning of this gene. Analysis of clinical data suggests a milder ADPKD phenotype for the PKD2 family. PMID- 9350816 TI - Evaluation of a project to enhance knowledge of hereditary diseases and management. AB - During 1992 and 1993, in a designated suburban area of Perth, Western Australia, information on hereditary disease was provided for health professionals and the general community. This information was in the form of posters, pamphlets, postal flyers and return letter cards, a static display, newspaper articles, advertisements and radio broadcasts, and professional seminars. The aim of this project was to evaluate the effectiveness of combined strategies to convey practical information about hereditary disease to the community and health professionals. Multiple measures of response evaluation were used, which included structured questionnaire surveys of health professionals and members of the community before and after the project. In the community surveys, respondents who were female, married, middle aged, and parents, and had a higher level of education or were born in Australia, New Zealand, or the United Kingdom were generally better informed about hereditary diseases. This intervention resulted in only meagre changes in community knowledge about hereditary disease, even though promotional materials were shown to be appropriate. General Practitioners (GPs) and Child Health Nurses (CHNs) were supportive of clinical genetic services and recognised a need for continuation of education in this field. There is a rapidly increasing need for community and health professional comprehension of the applications of the new genetic technology. This project indicates that routine educational and health promotion strategies will not be enough to achieve desired levels of knowledge and attitude change. PMID- 9350814 TI - Paternally inherited duplications of 11p15.5 and Beckwith-Wiedemann syndrome. AB - We present a three generation family in which a father and son have a balanced chromosome translocation between the short arms of chromosomes 5 and 11 (karyotype 46,XY,t(5;11)(p15.3;p15.3)). Two family members have inherited the unbalanced products of this translocation and are trisomic for chromosome 11p15.3 ->pter and monosomic for chromosome 5p15.3-->pter (karyotype 46,XY,der(5)t(5;11)(p15.3;p15.3)pat). Paternally derived duplications of 11p15.5 are associated with Beckwith-Wiedemann syndrome (BWS) and both family members trisomic for 11p15.5 had prenatal overgrowth (birth weights >97th centile), macroglossia, coarse facial features, and broad hands. We review the clinical features of BWS patients who have a paternally derived duplication of 11p15.5 and provide evidence for a distinct pattern of dysmorphic features in those with this chromosome duplication. Interestingly, our family is the fifth unrelated family to be reported with a balanced reciprocal translocation between the short arms of chromosomes 5 and 11. The apparently non-random nature of this particular chromosome translocation is suggestive of sequence homology between the two chromosome regions involved in the translocation. PMID- 9350817 TI - Wolfram (DIDMOAD) syndrome. AB - Wolfram syndrome (MIM 222300) is the association of juvenile onset diabetes mellitus and optic atrophy, also known as DIDMOAD (Diabetes Insipidus, Diabetes Mellitus, Optic Atrophy, and Deafness). Patients present with diabetes mellitus followed by optic atrophy in the first decade, cranial diabetes insipidus and sensorineural deafness in the second decade, dilated renal outflow tracts early in the third decade, and multiple neurological abnormalities early in the fourth decade. Other abnormalities include primary gonadal atrophy. Death occurs prematurely, often from respiratory failure associated with brainstem atrophy. Most patients eventually develop all complications of this progressive, neurodegenerative disorder. The pathogenesis is unknown, but the prevalence is 1 in 770000 in the UK and inheritance is autosomal recessive. A Wolfram gene has recently been mapped to chromosome 4p16.1, but there is evidence for locus heterogeneity, and it is still possible that a minority of patients may harbour a mitochondrial genome deletion. The best available diagnostic criteria are juvenile onset diabetes mellitus and optic atrophy, but there is a wide differential diagnosis which includes other causes of neurodegeneration. PMID- 9350818 TI - Marshall-Smith syndrome: the expanding phenotype. AB - We report a child of 3 years 9 months with the Marshall-Smith syndrome (MSS), characterised by the typical facial features, developmental delay, and advanced bone age. After the diagnosis was made at 5 months of age, careful observation for respiratory complications and failure to thrive was initiated. By 3 1/2 years of age, although our patient had no life threatening respiratory complications, investigation showed significant upper airway obstruction, which has been successfully treated. Aggressive treatment for failure to thrive has also allowed her to maintain a weight on the 50th centile. The purpose of this report is to suggest that early diagnosis and aggressive management may improve the ultimate prognosis with respect to the respiratory and feeding difficulties seen in this rare syndrome. PMID- 9350819 TI - Renal tubular dysgenesis with calvarial hypoplasia: report of two additional cases and review. AB - We report two cases of renal tubular dysgenesis (RTD) with calvarial hypoplasia and review the originally reported cases of RTD that came from our institution and published reports regarding the association of RTD and skull abnormalities. Although previously reported in association with RTD, calvarial hypoplasia is probably under-recognised. The cases reported here support the idea that the skull abnormalities observed in the inherited form of renal tubular dysgenesis are a common component of the disorder, as they are in the acquired form of RTD associated with maternal use of ACE inhibitors. Renewed attention to this clinical manifestation of RTD may be important in suggesting the diagnosis before death, providing more complete information to parents and physicians facing important management decisions and ensuring appropriate pathological examination postmortem. PMID- 9350821 TI - Parent-child transmission of infantile cholestasis with lymphoedema (Aagenaes syndrome). AB - We report a mother and daughter with features of Aagenaes syndrome. Unlike most previous cases, there is no Norwegian ancestry and the pedigree favours dominant rather than recessive inheritance. PMID- 9350820 TI - Macrocephaly, epilepsy, autism, dysmorphic features, and mental retardation in two sisters: a new autosomal recessive syndrome? AB - We report two sisters with macrocephaly, epilepsy, and severe mental retardation. The first child was a 14 year old girl born at term after a normal pregnancy, with birth weight 3600 g and occipitofrontal circumference (OFC) 36 cm (75th centile). Her head size increased markedly during the first six months of life, and was later stable at 2-3 cm above the 97.5th centile. Her development was characterised by psychomotor delay, epilepsy, and autistic features. Her face appeared mildly dysmorphic with a large forehead, short philtrum, and bushy eyebrows. Her younger sister was also born at term with birth weight 2600 g and OFC 34 cm (25th centile). She also developed postnatal macrocephaly with OFC 2 cm above the 97.5th centile and the same mild dysmorphic facial features as her sister. Her development was also characterised by psychomotor delay, autistic features, and epilepsy. In addition, she suffered from coeliac disease. She died unexpectedly at the age of 5 years, probably from an epileptic attack. Necropsy confirmed megalencephaly but no other pathological changes were found. The clinical features in these two sisters do not fit with any known syndrome and may represent a previously unrecognised autosomal recessive disorder. PMID- 9350822 TI - Spondylo-mesomelic-acrodysplasia with joint dislocations and severe combined immunodeficiency: a newly recognised immuno-osseous dysplasia. AB - A newborn girl is described with an association of spondylo-acrodysplasia, mild short limbed dwarfism without significant metaphyseal changes, joint dislocations, and severe immune system dysfunction. This association is distinct from other known immuno-osseous dysplasias, including Schimke dysplasia, ADA deficiency with osseous changes, and Omenn phenotype with short limbed dwarfism. PMID- 9350823 TI - The phenotypic effects of chromosome rearrangement involving bands 7q21.3 and 22q13.3. AB - We report a family in which the proband has a direct insertion of band 7q21.3 into chromosome 22 at 22q13.3, karyotype 46,XX,dir ins(22;7)(q13.3;q21.2q22.1). Two of her children have unbalanced chromosome rearrangements involving 7q21.3, with one girl monosomic for the region and a boy trisomic for the region. The child monosomic for band 7q21.3 has a split hand/split foot (SHSF) anomaly and her clinical features are consistent with the 7q21-q22 contiguous gene deletion syndrome. In situ hybridisation studies have shown that the proband and her son have a submicroscopic deletion of chromosome band 22q13.3. Interstitial deletions of this chromosome band have rarely been reported. PMID- 9350824 TI - Interstitial deletion of bands 4q12-->q13.1: case report and review of proximal 4q deletions. AB - We report a 6 year old child with a small de novo interstitial deletion of proximal 4q, karyotype 46,XX,del(4)(pter-->q12::q13.1-->qter). She has made good developmental progress and attends normal school with minimal assistance. We review published reports and clinical findings in patients with proximal 4q deletions. PMID- 9350825 TI - Isolated sacral agenesis in a fetus monosomic for 7q36.1-->qter. AB - A fetus with severe sacral agenesis and intrauterine growth retardation, ascertained at prenatal diagnosis, was found to be carrying an unbalanced form of a paternal balanced reciprocal translocation (7;19)(q36.1;q13.43), resulting in functional monosomy for 7q36.1-->qter. Necropsy confirmed that the fetus had isolated sacral agenesis type II. A critical region for autosomal dominant sacral agenesis has recently been mapped to the 7q36 region. This case provides further evidence for a sacral agenesis locus in this region and may help to refine the critical region further. PMID- 9350826 TI - Persistence of Mediterranean anaemia in Sicily. AB - We report 40 cases of homozygous beta thalassaemia, aged between 3 and 24 months, who were observed between January 1990 and June 1996 at the Thalassaemia Centre, Paediatric Department, Catania University. A questionnaire was used to find out the parents' knowledge of their risk before the birth of the affected children and showed that the persistence of Mediterranean anaemia in Sicily was mainly because of the following reasons: (1) poor information (62.5%), (2) laboratory error (12.5%), (3) difficulty in the differential diagnosis of beta thalassaemia trait (10%), and (4) not performing prenatal testing or selective abortion of an affected fetus (15%). We conclude that improved preventive measures at various medical and social levels can remove risk factors and so further reduce the incidence of Mediterranean anaemia in Sicily. PMID- 9350827 TI - Instability of normal (CTG)n alleles in the DM kinase gene. AB - We report on a myotonic dystrophy (DM) family exhibiting instability of normal sized (CTG)n alleles in the DM kinase gene on the non-DM chromosome. At least two mutational events involving normal DM alleles must have occurred in this family; one was characterised as a 34-35 (CTG)n repeat mutation. These findings represent a dissociation between (CTG)n repeat instability and myotonic dystrophy. Furthermore, this family highlights genetic counselling issues relating to the pathogenicity of alleles at the upper end of the normal size range and the risk of further expansion into the disease range. PMID- 9350828 TI - Mutation in the 3' region of the alpha-1-antitrypsin gene and chronic obstructive pulmonary disease. AB - A mutation in the 3' flanking region of the alpha-1-antitrypsin gene has been reported to be associated with chronic obstructive pulmonary disease (COPD). We have investigated the prevalence of this mutation in a group of 185 patients with airway obstruction and in 69 non-obstructed controls. The subjects were selected on the basis of their development of lung cancer and therefore had similar exposure to cigarette smoke, the major risk factor for COPD. In the majority of subjects, the level of airway inflammation and loss of elastic recoil was known, and therefore we were able to test whether the mutation was associated with one of these pathological mechanisms. The prevalence of heterozygotes for the mutation was 10% in both the obstructed group and controls. The mutation was not associated with increased airway inflammation or loss of elastic recoil. This result indicates that the 3' mutation is not a significant risk factor for COPD in this population, and suggests heterogeneity in the pathogenesis of the disease. PMID- 9350830 TI - Gaucher disease plus. PMID- 9350831 TI - Molecular characterisation of cystic fibrosis patients in the state of Sao Paulo (Brazil) PMID- 9350832 TI - Phorbol esters alter the morphology of cultured guinea-pig myenteric glia via a protein kinase C-independent mechanism. AB - Cultures of myenteric ganglia from adult guinea-pigs were used to study the influence of neuroactive substances on glial cells by monitoring changes in their morphology. The following substances had no effect on glial morphology: adenosine, ATP, carbachol, glutamate, bradykinin, isoprenaline, prostaglandin E2, sodium nitroprusside and lipopolysaccharide. The only substances found to affect glial morphology were phorbol esters, and in particular phorbol 12-myrisate 13 acetate (PMA), which acted at the nM range. Glial cells, which were normally polygonal, assumed a stellate shape within 30-60 min after the addition of PMA. Protein kinase C (PKC) inhibitors did not block this effect, and PKC activators did not mimic it. The effect of PMA was also not mediated by changes in the intracellular concentrations of either Ca2+, H+ or cyclic AMP. Dye coupling among glial cells was blocked by PMA. The phorbol ester-mediated effect on glial structure may have profound influence on neuronal organization and function. PMID- 9350833 TI - Sexual impotence is associated with a reduced production of oxytocin and with an increased production of opioid peptides in the paraventricular nucleus of male rats. AB - Oxytocin plays a physiological stimulatory role on sexual behavior. Conversely, opioid neuropeptides play a physiological inhibitory role. Here we show that in sexually impotent rats there is a reduced expression of oxytocin mRNA and an increased expression of proenkephalin and pro-dynorphin mRNA in the paraventricular nucleus of hypothalamus (PVN), a brain structure of key importance for sexual behavior. These data suggest that an imbalance in the production of oxytocin and of opioid peptides in the PVN, with prevalence of opioid peptides, may underlie a condition of sexual impotence. PMID- 9350834 TI - Carbachol-induced inositol phosphate formation in rat striatum is mediated by both M1 and M3 muscarinic receptors. AB - In vibratome-cut slices from rat striatum and in the presence of 10 mM LiCl, the cholinergic agonist carbachol stimulated the accumulation of total [3H]inositol phosphates (EC50 11+/-1 microM and maximum effect 546+/-36% of basal). The response to 100 microM carbachol (497+/-24% of basal) was inhibited by muscarinic antagonists (1 microM), the rank order of efficacy being 4-diphenylacetoxy-N methylpiperidine methiodide (4-DAMP; 100% inhibition) approximately pirenzepine (98+/-3%) > p-fluoro analog of hexahydro-sila-difenidol (pFHHSiD; 90+/-3%) >> methoctramine (32+/-7%) approximately tropicamide (30+/-10%). Antagonist inhibition curves best fit to a single-site model for 4-DAMP (pKi 8.9+/-0.2) whereas, for both pirenzepine and pFHHSiD, the best fit was to the two-site model. The pKi values for the high-affinity (8.3+/-0.2) and low-affinity (6.9+/ 0.2) components for pirenzepine-mediated inhibition corresponded to those reported for M1 and M3 receptors, respectively. The pKi values for the high affinity (8.2+/-0.3) and low-affinity (7.0+/-0.2) components for pFHHSiD inhibition were in good agreement with those reported for M3 and M1 receptors, respectively. Altogether these results indicate that carbachol-induced [3H]inositol phosphate formation in rat striatal slices is mediated by both M1 and M3 muscarinic receptors. PMID- 9350835 TI - Growth/differentiation factor 5 protects nigrostriatal dopaminergic neurones in a rat model of Parkinson's disease. AB - Growth/differentiation factor 5 (GDF5), a novel member of the transforming growth factor beta superfamily, promotes the survival of dopaminergic neurones in vitro. We present here the first evidence for a neuroprotective action of GDF5 in vivo. We investigated the effects of intracerebral administration of GDF5 on a rat model of Parkinson's disease. GDF5 was administered just above the substantia nigra and into the lateral ventricle immediately before ipsilateral injection of 6-hydroxydopamine into the medial forebrain bundle. GDF5 prevented the development of amphetamine-induced rotations and preserved the integrity of striatal dopaminergic nerve terminals, as measured by positron emission tomography. Post-mortem studies showed that GDF5 spared dopamine levels in the striatum and tyrosine hydroxylase positive neurones in the midbrain. This study suggests that GDF5 has potential for the treatment of Parkinson's disease. PMID- 9350836 TI - Effects of thermal acclimation on the neurotransmitters, serotonin and norepinephrine in the discrete brain of male and female tilapia, Oreochromis mossambicus. AB - Effects of thermal acclimation on the serotonin (5-HT) and norepinephrine (NE) contents in the discrete brain of male and female tilapia, Oreochromis mossambicus were investigated. Sexually mature males and females were exposed to 26 degrees C, 29 degrees C, or 32 degrees C of water temperature for 3 weeks. The hypothalamic 5-HT content in the 29 degrees C and 32 degrees C acclimated male was lower than that in the 26 degrees C group. In females, the hypothalamic 5-HT content in the 32 degrees C acclimated group was less than those in the 26 degrees C and 29 degrees C groups. Similar results were found in the hypothalamic NE contents of males and females. In the optic lobe, the elevated temperature acclimation (29 degrees C and 32 degrees C) resulted in a higher 5-HT content in both males and females; whereas, the NE content was increased by the elevated temperature acclimation in females but not altered in that of males. In the telencephalon, the elevated temperature acclimation had no influence on the 5-HT content of males and females, but resulted in a lower NE content in both males and females. These results demonstrate that the neurotransmitter activity of teleost is influenced by the thermal acclimation in a sex- and regional-dependent pattern. The alterations of 5-HT and NE in the central nervous system might be involved in the physiological and biochemical responses that occur during thermal acclimation in fish. PMID- 9350837 TI - Long duration pressor responses following activation of subfornical organ neurons in rats are the result of increased circulating vasopressin. AB - Electrical stimulation in the subfornical organ (SFO) of male Sprague-Dawley rats resulted in biphasic increases in blood pressure (BP) without a change in heart rate. The initial short duration (0-10 s) increase in BP lasted throughout the 10 s stimulation period (area under the curve (AUC) = 104.3+/-15.26 mmHg/s, (mean+/ SEM) P < 0.001). Upon termination of the electrical stimulus, the BP remained elevated for approximately 55 s (long duration response, AUC = 327.5+/-48.22 mmHg/s, P < 0.001). This long duration BP response was determined to be the result of an increase in circulating vasopressin (VP) as administration of a V1 receptor antagonist abolished this response (AUC = -210.7 +/- 42.38 mmHg/s, P < 0.01). The results of the present study demonstrate that the long duration component of the biphasic increase in BP observed on response to electrical stimulation of the SFO is the result of increased concentrations of circulating VP. PMID- 9350838 TI - Basic fibroblast growth factor-induced seizures in rats. AB - The neurotrophic effects of basic fibroblast growth factor on hippocampal neurons have been well documented. However, the acute effects of basic fibroblast growth factor on hippocampal electrical activities in vivo have not yet been studied. To assess the impact of basic fibroblast growth factor on hippocampal electroencephalographic (EEG) activity, two doses of basic fibroblast growth factor were injected into the right ventral hippocampus of freely moving animals. Unilateral injection of 50 ng of basic fibroblast growth factor into the dentate region of the hippocampus resulted in immediate EEG ictal discharges and behavioral seizures in 7/9 rats. These seizures lasted about 2 h following the injection. Injection of 25 ng of basic fibroblast growth factor into the same sites induced ictal discharges of approximately 60 min duration in 3/7 rats. Injection with heat-inactivated basic fibroblast growth factor into the same sites did not cause any EEG or behavioral modifications. These results suggest that in addition to growth-promoting effects, basic fibroblast growth factor is able to acutely alter the electrical activity of hippocampal neurons and cause seizures. PMID- 9350839 TI - Morphologic difference of neuropil threads in Alzheimer's disease, corticobasal degeneration and progressive supranuclear palsy: a morphometric study. AB - Using Gallyas-Braak's silver stain, neuropil threads (NTs) in Alzheimer's disease (AD), corticobasal degeneration (CBD) and progressive supranuclear palsy (PSP) were analyzed morphologically and morphometrically. The NT density was highest in the cerebral cortical layer V in AD and CBD, and in the subcortical white matter in PSP. An overlaid, two-dimensional, camera lucida drawing revealed differences in the fine profile of NTs among these three disease groups. The differences were confirmed by computerized feature analysis which revealed differences in maximum length, breadth, Feret's angle and orientation of the NTs. Unlike a previous assumption that all NTs have a similar appearance, our study revealed that NTs in AD, CBD and PSP were distinctively different with respect to their morphology. PMID- 9350840 TI - The role of nitric oxide in the control of plasma glucose concentration in spontaneously hypertensive rats. AB - Glucose homeostasis was studied in the spontaneously hypertensive rat (SHR). The fasting plasma glucose levels were similar in the SHR and normotensive Wistar Kyoto (WKY) rat (102.7+/-2.4 vs. 107.4+/-4.2 mg/dl, P > 0.01). One hour after glucose challenge, the plasma glucose level was slightly but insignificantly increased in both SHR and WKY rat (117+/-2.5 vs. 114.3+/-3.2 mg/dl, P > 0.01). After N(G)nitro-L-arginine methyl ester (L-NAME) 20 mg/kg per day was administered intraperitoneally (i.p.) for 4 days, the plasma glucose level was significantly increased in the rats (SHR 167.3+/-4.9; WKY rat 136.0+/-4.8 mg/dl); the increase was significantly more pronounced in the SHR. The fasting insulin levels were similar in the SHR and WKY rats (2.3+/-0.4 vs. 2.0+/-0.3 ng/ml, P > 0.01). One hour after glucose challenge, the insulin level was significantly increased in the WKY rat (4.8+/-0.7 ng/ml) but not in the SHR (2.2+/-0.4 ng/ml). With L-NAME treatment, plasma insulin increase was noted in the WKY rat but not SHR (4.6+/-0.6 vs. 2.6+/-0.4 ng/ml, n = 8, P < 0.01). One hour after insulin 1 IU/kg was injected intramuscularly (i.m.), the plasma glucose level was significantly decreased in both the SHR (from 115.0+/-6.5 to 48.6+/-3.6 mg/dl, n = 8) and WKY rat (from 108.3+/-3.8 to 52.6+/-4.2 mg/dl, n = 8). No significant difference was noted between the decrease of the two groups (P > 0.01). The present findings suggested that NO plays a role in the glucose homeostasis of rats. NO-synthase blockade resulted in an increase of plasma glucose level. The SHR maintains normal glucose level and tolerance in spite of a defective insulin release response. This is probably due the compensatory effect of a more prominent NO-dependent glucose homeostatic function. PMID- 9350841 TI - Role of phospholipase A2 in the release of free fatty acids during ischemia reperfusion in the rat cerebral cortex. AB - Phospholipases are implicated in ischemic damage. In this study, we have examined the time course of changes in free fatty acid (FFA) levels and cytosolic phospholipase A2 (PLA2) activity during 10 or 20 min periods of four vessel occlusion rat cerebral ischemia followed by reperfusion. Ischemia for a duration of 20 min resulted in a biphasic release of FFA in rat cortical superfusates. There was a rapid elevation in the first 10 min followed by a slower release in the next 10 min. Reperfusion for 10 min resulted in another rapid release of FFA and thereafter the levels gradually declined, but did not return to basal levels. Measurements by enzymatic assay and Western blot analysis showed a significant increase in the activity and protein levels of cPLA2 during the ischemic periods. These remained elevated at 10 min of reperfusion, but returned to base line levels after 20 min of reperfusion. The possible role of phospholipase A2 (PLA2) in the release of FFA and ischemic injury to the brain is discussed. PMID- 9350842 TI - Developmental lead exposure causes spatial learning deficits in adult rats. AB - Groups of male rats exposed to lead (Pb) during different developmental periods were tested as adults in a water maze. A highly significant (P < 0.01) impairment in water maze performance was measured in rats exposed to Pb only during gestation and lactation (maternal exposure). At the time of testing (100-106 days old), blood and brain Pb concentrations were at control levels. Significant impairments (P < 0.05) were also present in rats continuously exposed to Pb from conception through adulthood. Post-weaning Pb exposure alone did not result in impaired performance despite significantly elevated blood and brain Pb levels at the time of testing. This study supports the hypothesis that a window of vulnerability to Pb neurotoxicity exists in the developing brain and that Pb exposure can result in long-term cognitive deficits. PMID- 9350843 TI - Quantification of cat's articular afferents containing calcitonin gene-related peptide or substance P innervating normal and acutely inflamed knee joints. AB - In cats with an acute (32 h) unilateral knee joint inflammation the proportion of calcitonin gene-related peptide-(CGRP) and substance P-(SP) immunoreactive articular afferents, retrogradely labelled by Fast Blue (FB), were determined using immunohistochemistry. The proportion of neurons containing CGRP was significantly higher on the inflamed side (52%) than on the contralateral side (39%) and in controls (42%). However, the proportion of SP-immunoreactive articular perikarya on the inflamed side (26%) did not differ from the contralateral side (24%) and the control cats (22%). These data indicate that acute inflammation induces the synthesis of CGRP but not of SP in joint afferents. PMID- 9350844 TI - The pontine micturition center projects to sacral cord GABA immunoreactive neurons in the cat. AB - Stimulation of the pontine micturition center (PMC) results in micturition, i.e. an immediate relaxation of the bladder sphincter and a contraction of the detrusor muscle of the bladder. Earlier studies have shown that the bladder contraction is brought about by a direct excitatory pathway from the PMC to the parasympathetic bladder motoneurons in the sacral cord. How the PMC produces the inhibition of the bladder sphincter is not known. The present study in two adult male cats demonstrates at the ultrastructural level a direct pathway from the PMC to the dorsal gray commissure of the sacral cord. More than half (55%) of these terminals made contact with gamma amino butyric acid (GABA) immunoreactive neurons or somata, the others with non-GABA immunoreactive profiles. The PMC terminals contained many round vesicles, some dense cored vesicles and exclusively asymmetric synaptic clefts, which correspond with an excitatory pathway. A concept is put forward in which this pathway produces the relaxation of the bladder sphincter during micturition. PMID- 9350845 TI - The reorganization of mu opioid receptors in the rat dorsal horn following peripheral axotomy. AB - The mu opioid receptor is concentrated in laminae I and II (LI and LII, respectively) of the normal rat dorsal horn. Fourteen days after transection of the L4-L6 segmental peripheral nerves, image analysis demonstrates a 49, 34 and 17% decrease in mu opioid staining density in the medial, middle and lateral thirds of the superficial dorsal horn, respectively, when comparing the operated to the unoperated side. Intralaminar analysis demonstrates that the greatest change in density occurs in LI and LII outer, compared to LII inner. By 31 days post-surgery, staining has returned to normal with side to side differences no longer present. These results imply that mu opioid ligands such as morphine might be less effective in ameliorating pain 2 weeks after a peripheral nerve lesion than they are in the normal condition, but that this effectiveness should return as the receptors are restored to their normal levels. Thus, the time following a lesion may be an important variable in assessing the effectiveness of mu opioid ligands in alleviating neuropathic pain. Furthermore, this study shows that the organization of opioid receptors in the superficial dorsal horn is malleable and could lead to changes in drug efficacy. PMID- 9350846 TI - The time course of interhemispheric EEG coherence during a GO/NO-GO task in humans. AB - Event-related coherence of the EEG was calculated for 10 subjects performing a visual discrimination GO/NO-GO task. The subjects were instructed to push (GO) or not to push (NO-GO) a button according to visual stimuli. Twenty-one-channel scalp EEGs were recorded and the surface Laplacian was calculated using the source derivation method. The time courses of the coherence between F3 and F4, C3 and C4, and P3 and P4 were calculated using the fast Fourier transform for each task and were compared between conditions. Statistical analysis showed that coherence in the NO-GO condition became significantly higher than that in the GO condition between F3 and F4. The synchronization between bilateral dorsolateral frontal areas might therefore play an important role in the motor inhibition process. PMID- 9350847 TI - Chronic psychosocial stress does not affect the number of pyramidal neurons in tree shrew hippocampus. AB - Exposure of the hippocampus for prolonged periods to an excess of glucocorticoids is thought to result in damage and finally loss of pyramidal neurons. By applying improved methods for quantifying cell numbers we investigated whether chronic psychosocial stress in tree shrews reduces the total number of hippocampal pyramidal neurons. For 28 days male tree shrews were exposed to subordination stress resulting in constantly elevated cortisol levels. The number of pyramidal neurons in the hippocampal fields CA1 and CA3 was estimated with the optical fractionator technique. In both hippocampal fields, neuron number in the stressed subjects was not significantly altered in comparison to unstressed controls. This constancy of neuron number represents an essential finding for identifying the effects of chronic stressful conditions on hippocampal structure. PMID- 9350848 TI - Expression of mRNAs for neuropeptide receptors and beta-adrenergic receptors in human osteoblasts and human osteogenic sarcoma cells. AB - In human periosteum-derived osteoblastic cells (SaM-1) and human osteosarcoma derived cells (SaOS-2, HOS, MG-63), the mRNA expressions of calcitonin gene related peptide receptor (CGRP-R), substance P receptor (SP-R), neuropeptide Y receptor (NPY-R), beta-adrenergic receptors (beta1-R, beta2-R, beta3-R), vasoactive intestinal polypeptide type 1 and type 2 receptors (VIP-1R, VIP-2R) and pituitary adenylate cyclase activating polypeptide receptor (PACAP-R) were examined by reverse transcription-polymerase chain reaction (RT-PCR). According to the magnitude of the mRNA expression of alkaline phosphatase (ALP), the relative state of commitment of these osteoblastic cell lines to the osteoblast lineage was SaM-1 > SaOS-2 > HOS > MG-63. CGRP-R, NPY-R, VIP-1R and beta2-R, but not SP-R, VIP-2R, PACAP-R, beta1-R and beta3-R, were expressed in osteoblasts as well as osteosarcoma cells. Expression of these receptors seems to be a common feature in osteoblastic cells, but the magnitude of expression was not dependent upon the relative state of commitment of the osteoblastic cells to the osteoblast lineage. In addition, VIP mRNA was not expressed in osteoblastic cells, suggesting the absence of an autocrine system of VIP in osteoblasts. These observations suggest that these neuropeptides and norepinephrine are involved in local regulation of human bone metabolism. PMID- 9350849 TI - Blockage of urinary responses by inhibitors for IP3-mediated pathway in rat vomeronasal sensory neurons. AB - The mammalian vomeronasal system is involved in the effects of urinary chemicals on gonadal functions and sexual behaviors. For example, exposure to urine affects the timing of oestrous cycles in rats. Rat vomeronasal sensory neurons in slice preparation were studied under on-cell patch clamp conditions. We found that urine excreted from male Wistar rats increased impulse frequency in vomeronasal sensory neurons of female Wistar rats. The urinary responses were blocked by an inositol-1,4,5-trisphosphate (IP3)-channel inhibitor (10 microM ruthenium red) or phospholipase C inhibitors (10 microM U-73122 and 1 mM neomycin), suggesting that pheromone-like substances in the urine induce the response in the rat vomeronasal sensory neurons via the IP3-dependent transduction pathway. PMID- 9350850 TI - Nitric oxide does not modulate kainate receptor binding in human brain. AB - The ability of three nitric oxide (NO) donor compounds to modify ligand binding to kainate receptors was studied in tissue from human adult autopsy brains. Binding of [3H]kainic acid (5 nM) was measured in frontal cortex membranes made from Brodmann area 8 (BA 8) and autoradiographically using sections of frontal cortex (BA 8 and 9). None of the three donors, S-nitroso-N-acetyl-D,L penicillamine (SNAP), S-nitrosocysteine (Cys-NO) and 3-morpholinosydnonimine chloride (SIN-1) altered the specific binding of [3H]kainic acid. Nitrite accumulation assays confirmed that adequate amounts of NO were released by the donors under the ligand binding conditions used. The findings show that binding to the kainate receptor, in contrast to the other ionotropic glutamate receptors, is not affected by NO and strongly suggest that endogenous NO produced by NO synthase (NOS) does not modulate kainate receptors in vivo. Mechanisms whereby NOS inhibitors potentiate kainic acid-induced seizures in animal models may include altered modulation of glutamate N-methyl-D-aspartate (NMDA) receptors. PMID- 9350851 TI - A late ON response remains in visual response of the mGluR6-deficient mouse. AB - In a previous study, we showed that the visual ON response recorded from the superior colliculus is absent in the metabotropic glutamate receptor subtype 6 (mGluR6) knockout mice. However, these mutant mice can respond to some visual inputs and are able to learn a light-dark discrimination task. Here, we systematically investigated the possibility that some ON responses could remain in the mutant mice. Recording light evoked field potentials from the superior colliculus of mGluR6-deficient mice at scotopic and mesopic backgrounds, we found a characteristic ON response that differs from the ON response of wild-type mice. The ON responses recorded from the mutant mice appeared with a latency (200-300 ms) longer than that of the wild-type mice (50-100 ms) and amplitudes of this late ON response decreased upon increase of the light intensity in most cases examined. The late ON response may contribute to the apparent normal behavior and some physiological responses to light stimuli in mGluR6-deficient mice. PMID- 9350852 TI - Sigma ligands stimulate GTPase activity in mouse prefrontal membranes: evidence for the existence of metabotropic sigma receptor. AB - We studied effects of various sigma ligands on GTPase activity in mouse prefrontal membranes. Some representative sigma agonists, such as (+) pentazocine, SA4503 and (+)-3-PPP, stimulated the GTPase activity in a concentration-dependent manner in ranges of 10 nM to 10 microM. Maximal effect was almost 10% increase to the control without treatment of drugs. However, another representative agonist, (+)-SKF10,047 showed only a partial activity with maximal effect 5% at 1 microM. NE-100, a representative antagonist, showed no effect at concentrations not more than 100 nM, while it did stimulate GTPase activity at 1 and 10 microM. Furthermore, these stimulative effects of both (+) pentazocine and SA4503 on GTPase activity were significantly antagonized by NE 100 at 100 nM, suggesting that NE-100 possesses agonist-antagonist property. These findings suggest the possibility that there exist metabotropic sigma receptors. PMID- 9350853 TI - No association between the low density lipoprotein receptor-related protein (LRP) gene and late-onset Alzheimer's disease in a community-based sample. AB - It is now commonly known that possession of the epsilon4 allele of the apolipoprotein E (APOE) gene confers an increased risk for both familial and sporadic Alzheimer's disease (AD), in a dose-dependent way. Other genes that may play a role in AD, either through independent association with the disease or through modification of the existing APOE risk, have been reported with conflicting results. One such gene, the low density lipoprotein receptor-related protein (LRP) gene, was recently reported by two groups to be associated with AD, although the groups identified different risk-conferring alleles. Both studies were based on clinic-derived AD populations (one American, one French), and both reported only marginally significant results. We have genotyped a community-based AD and control population at this LRP polymorphism and find no association between the variants at that polymorphism and the occurrence of AD. Further, despite the biochemical relationship between LRP and the ApoE protein, we find no significant statistical interaction between the alleles at these loci. PMID- 9350854 TI - Fast blue, a fluorescent tracer in glioma cell culture, affects cell proliferation and motility. AB - The azo-dye, Fast Blue (FB), initially employed for retrograde neuronal tracing is increasingly used in cell invasion and migration studies to detect living cells in monolayer and glioma tumor cell spheroid models. As yet, it is assumed that a cell stained with a tracker dye retains the characteristics of the original cell. The following experiments compared the adhesion, migration and proliferation properties of the cell lines U373 and GaMG with and without FB staining. FB staining reduced cell adhesion (P < 0.01) and proliferative activity (P < 0.01) and also had a significant inhibitory effect on cell migration (P < 0.001). From the results presented it follows that FB staining markedly influences basic cell characteristics. PMID- 9350856 TI - Subdivisions of the olfactory system in the sterlet Acipenser ruthenus. AB - Subdivisions of the olfactory system of the sterlet Acipenser ruthenus were investigated by means of horseradish peroxidase (HRP) injections into the nose, and by soybean agglutinin binding studies. With both methods primary olfactory fibers were labeled which projected to the ventral part of the glomerular layer of the olfactory bulb. The dorsal part of the olfactory bulb did not bind soybean agglutinin, however, even though HRP tracing showed primary olfactory fibers in that area. This confirms earlier morphological studies which claim the existence of distinct subdivisions of the olfactory system in the sturgeon. The lack of soybean agglutinin binding in the dorsal part of the olfactory bulb suggests, however, that this part is not homologous with the accessory olfactory system of tetrapods. PMID- 9350855 TI - Dissociation between the perception of body verticality and the visual vertical in acute peripheral vestibular disorder in humans. AB - Estimates of the subjective visual and postural vertical were obtained from five patients with acute peripheral vestibular lesions and 20 normal subjects. The visual vertical was assessed by asking the subjects to align a target line to earth vertical by means of remote control. Postural vertical judgments were obtained by exposing them to rotational displacements in the roll plane while sitting on a motor-driven chair and requiring them to align their body to vertical using a joystick control. While the patients showed strong deviations of the visual vertical towards the lesion side, their postural vertical judgments remained veridical. We conclude that the above perceptions are not processed identically and that the participating sensory systems are differently weighted during these tasks. PMID- 9350857 TI - Real-time measurement of nitric oxide production in rat brain by the combination of luminol-H2O2 chemiluminescence and microdialysis. AB - A chemiluminescence method of detecting nitric oxide (NO) in combination with a microdialysis technique was employed for the real-time measurement of NO production in living rat brain. This method based on the luminol-H2O2 system has a detection limit of 1 nM, and is the most sensitive method currently available for measuring NO. We applied this new technique to rat cerebellum to record the increase of chemiluminescence intensity arising from NO production after the injection of N-methyl-D-aspartate or kainic acid around the microdialysis probe. This highly sensitive method should be useful for the direct clarification of the functions of NO in the central nervous system. PMID- 9350858 TI - Control of the ToxR virulence regulon in Vibrio cholerae by environmental stimuli. AB - Many bacterial pathogens regulate the expression of virulence genes in a co ordinate manner in response to changes in the environment. For example, the human pathogen, Vibrio cholerae, possesses a virulence regulon composed of over 20 genes involved in colonization, toxin production and bacterial survival within the host, which are co-ordinately regulated by external stimuli, such as temperature, pH and osmolarity. Although the expression of the regulon is dependent upon the transcriptional activator ToxR, most of these genes are controlled by a second transcriptional activator, ToxT, which is itself positively regulated by ToxR. The mechanisms by which environmental stimuli influence the ToxR regulon are not yet understood, but ToxR-mediated control over the expression of toxT clearly plays a role. The recent finding that the global regulator cAMP-CRP also influences the expression of the ToxR regulon under various environmental conditions raises new issues regarding the pathways and mechanisms by which this regulation is achieved and indicates that multiple overlapping systems are involved. PMID- 9350859 TI - Nicking by transesterification: the reaction catalysed by a relaxase. AB - DNA relaxases play an essential role in the initiation and termination of conjugative DNA transfer. Purification and characterization of relaxases from several plasmids has revealed the reaction mechanism: relaxases nick duplex DNA in a site- and strand-specific manner by catalysing a transesterification. The product of the reaction is a nicked double-stranded DNA molecule with a sequestered 3'-OH and the relaxase covalently bound to the 5' end of the cleaved strand via a phosphotyrosyl linkage. The relaxase-catalysed transesterification is isoenergetic and reversible; a second transesterification ligates the nicked DNA. However, the covalent nucleoprotein complex is relatively long-lived, a property that is likely to be essential for its role as an intermediate in the process of conjugative DNA transfer. Subsequent unwinding of the nicked DNA intermediate is required to produce the single strand of DNA transferred to the recipient cell. This reaction is catalysed by a DNA helicase, an activity intrinsic to the relaxase protein in some, but not all, plasmid systems. The first relaxase-catalysed transesterification is essential for initiation of conjugative strand transfer, whereas the second is presumably required for termination of the process. The relaxase, in conjunction with several auxiliary proteins, forms the relaxation complex or relaxosome first described nearly 30 years ago as being associated with conjugative and mobilizable plasmids. PMID- 9350860 TI - Altered ParA partition proteins of plasmid P1 act via the partition site to block plasmid propagation. AB - The partition system of the P1 plasmid, P1par, consists of the ParA and ParB proteins and a cis-acting site, parS. It is responsible for the orderly segregation of plasmid copies to daughter cells. Plasmids with null mutations in parA or parB replicate normally, but missegregate. ParB binds specifically to the parS site, but the role of ParA and its ATPase activity in partition is unclear. We describe a novel class of parA mutants that cannot be established or maintained as plasmids unless complemented by the wild-type gene. One, parAM314l, is conditional: it can be maintained in cells in minimal medium but cannot be established in cells growing in L broth. The lack of plasmid propagation in L broth-grown cells was shown to be caused by a ParB-dependent activity of the mutant ParA protein that blocks plasmid propagation by an interaction at the parS site. Thus, ParA acts to modify the ParB-parS complex, probably by binding to it. Partition is thought to involve selection of pairs of plasmids before segregation, either by physical pairing of copies or by binding of copies to paired host sites. We suggest that ParA is involved in this reaction and that the mutant ParA protein forms paired complexes that cannot unpair. PMID- 9350861 TI - Mutational analysis of slyD, an Escherichia coli gene encoding a protein of the FKBP immunophilin family. AB - slyD encodes a 196 amino acid polypeptide that is a member of the FKBP family of cis-trans peptidyl-prolyl isomerases (PPlases). slyD mutations affect plaque formation by the phage phiX174 by blocking the action of the phage lysis protein E. Here we describe the selection of a set of spontaneous slyD mutations conferring resistance to the expression of gene E from a plasmid. These mutations occur disproportionately in residues of SlyD that, based on the structure of the prototype mammalian FKBP12, make ligand contacts with immunosuppressing drug molecules or are conserved in other FKBP proteins. A wide variation in the plating efficiency of phiX174 on these E(R) strains is observed, relative to the parental, indicating that these alleles differ widely in residual SlyD activity. Moreover, it is found that slyD mutations cause significant growth rate defects in Escherichia coli B and C backgrounds. Finally, overexpression of slyD causes filamentation of the host. Thus, among the FKBP genes found in organisms across the evolutionary spectrum, slyD is unique in having three distinct drug independent phenotypes. PMID- 9350862 TI - Clonal descent and microevolution of Neisseria meningitidis during 30 years of epidemic spread. AB - Serogroup A meningococci of subgroups III, IV-1 and IV-2 are probably descended from a common ancestor that existed in the nineteenth century. The 10.5kb sequences spanning five distinct chromosomal loci, encoding cell-surface antigens, a secreted protease or housekeeping genes and intergenic regions, were almost identical in strains of those subgroups isolated in 1966, 1966 and 1917 respectively. During the subsequent two to three decades, all of these loci varied as a result of mutation, translocation or import of DNA from unrelated neisseriae. Thus, microevolution occurs frequently in naturally transformable bacteria. Many variants were isolated only once or within a single geographical location and disappeared thereafter. Other variants achieved genetic fixation within months or a few years. The speed with which sequence variation is either eliminated or fixed may reflect sequential bottlenecks associated with epidemic spread and contrasts with the results of phylogenetic analyses from bacteria that do not cause epidemics. PMID- 9350863 TI - The dipeptide repeat region of the fibrinogen-binding protein (clumping factor) is required for functional expression of the fibrinogen-binding domain on the Staphylococcus aureus cell surface. AB - Clumping factor of Staphylococcus aureus is a fibrinogen-binding protein that is located on the bacterial cell surface. The protein has an unusual repeat domain (region R) comprising mainly the dipeptide aspartate and serine. To determine if region R has a role in the surface display of the fibrinogen-binding region A domain, deletions lacking the region R encoding region of the clfA gene were generated. To determine the minimum length of region R required for wild-type levels of ClfA expression, variants with truncated region R domains were constructed. S. aureus cells expressing mutated clfA genes were tested for (i) proteins released by lysostaphin treatment that reacted with antisera specific for region A, (ii) clumping in soluble fibrinogen, (iii) adherence to immobilized fibrinogen and (iv) expression of the ClfA antigen on the cell surface by fluorescent activated cell sorting analysis. Each construct expressed three major immunoreactive proteins, two of which were putative N-terminal degradation products. Region R residues greater than 40 were required between region A and W (72 residues between region A and the LPDTG sorting signal) for wild-type levels of clumping in fibrinogen. A stepwise decrease in clumping titre was observed as the distance between region A and LPDTG was decreased from 72 to 4 residues. Similarly, a decrease in binding of anti-ClfA serum and in binding to fibrinogen coated plastic surfaces was observed with cells expressing ClfA with 40 region R residues or less. Nevertheless, low levels of adherence to fibrinogen and binding to anti-ClfA serum occurred with ClfA derivatives that lacked region R altogether. This indicates that a small proportion of the ClfA molecules are linked to peptidoglycan very close to the cell surface but that residues greater than 72 are needed to allow sufficient ClfA molecules to span the entire cell wall and to display the biologically active A domain in a form that can participate fully in fibrinogen binding. PMID- 9350864 TI - A developmentally regulated Streptomyces endoribonuclease resembles ribonuclease E of Escherichia coli. AB - We report that the Streptomyces species S. lividans and S. coelicolor, morphologically complex gram-positive soil bacteria, contain a developmentally regulated endoribonuclease activity (here named RNase ES) that functionally and immunologically resembles Escherichia coli RNase E. In Streptomyces cells, RNA I the antisense repressor of replication of ColE1-type plasmids - is cleaved at sites attacked by RNase E. A Mg2+-dependent endonuclease that produces RNase E like cleavages in RNA I and 9S ribosomal RNA was identified in S. lividans cell extracts. A Streptomyces peptide migrating at 70kDa in SDS/polyacrylamide gels binds to RNase E substrates and reacts with three separate anti-RNase E monoclonal antibodies; the endonucleolytic cleavage activity co-purified with the immunoreactive 70 kDa peptide. We show that RNase ES activity is regulated during the Streptomyces life cycle: activity increased as cells progressed from exponential growth to stationary phase in liquid culture, or from mycelial growth to sporulation on solid media. While mutations that interfere with S. coelicolor development late in its life cycle did not prevent this developmentally associated increase in RNase ES activity, the increase was blocked by a mutation (bldA) that interferes early with both morphological and physiological differentiation. PMID- 9350865 TI - The division during bacterial sporulation is symmetrically located in Sporosarcina ureae. AB - Immunofluorescence microscopy was used to visualize the FtsZ band that marks the site of septation in Sporosarcina ureae. Image analysis indicated that the vegetative division was symmetrically located with respect to the ends of the cells. Fusions of lacZ to the sporulation loci, spollA and cotE, of Bacillus subtilis were introduced into S. ureae by mobilization of plasmids containing the fusions from Escherichia coli. The fusions showed similar patterns of sporulation associated expression in S. ureae to those observed in B. subtilis. Formation of beta-galactosidase encoded by the spollA-lacZ fusion made it possible to identify early sporulating cells by immunofluorescence microscopy. Analysis of the position of FtsZ bands in cells expressing spollA-lacZ indicated that the location of sporulation division was symmetrical with respect to the ends of the cells, in sharp contrast to the asymmetrical location of septation in sporulating Bacilli. It is inferred that asymmetry of location of the sporulation division is not essential for the compartmentalization of gene expression that follows the division. PMID- 9350866 TI - Phase variation in tcpH modulates expression of the ToxR regulon in Vibrio cholerae. AB - We evaluated a spontaneous mutant of Vibrio cholerae, which was avirulent in an infant mouse and had reduced expression of cholera toxin and TcpA in response to environmental signals. The toxR, toxS and toxT genes in the mutant were normal, but transcription of toxT was absent. A plasmid expressing wild-type tcpP and tcpH complemented the mutant. The mutation resulted from a frameshift in a string of nine G residues within tcpH; similar slipped-strand mutations in tcpH arose at a frequency of 10(-4) during overnight growth and in the majority of colonies by the end of 5 days of growth in ToxR-inducing conditions. Transcription of tcpPH was regulated by temperature and pH independently of ToxR or ToxT. These results suggest that TcpH couples environmental signals (temperature and pH) to expression of the ToxR regulon, and provide a model for phase variation in the co ordinate expression of cholera virulence factors. PMID- 9350867 TI - SpsA, a novel pneumococcal surface protein with specific binding to secretory immunoglobulin A and secretory component. AB - The interaction of pathogenic bacteria with host serum and matrix proteins is a common strategy to enhance their virulence. Streptococcus pneumoniae colonizes the human upper respiratory tract in healthy individuals and is also able to cause invasive diseases. Here, we describe a novel pneumococcal surface protein, SpsA, capable of binding specifically to human secretory immunoglobulin A (SIgA). The dissociation constant of SIgA binding to SpsA was 9.3 x 10(-9) M. Free secretory component (SC) also binds to S. pneumoniae, whereas serum IgA does not, suggesting that pneumococcal binding to SIgA is mediated by the SC. To our knowledge, this is the first defined interaction of SC with a prokaryotic protein. The spsA gene encodes a polypeptide of 523 amino acids with a predicted molecular mass of 59 151 Da. The SIgA- or SC-binding domain is located in the N terminal part of SpsA and exhibits no significant homology to any other proteins. The purified SIgA-binding domain of SpsA could completely inhibit the binding of SIgA to pneumococci. SpsA was expressed by 73% of the tested S. pneumoniae isolates and was substantially conserved between different serotypes. The interaction between S. pneumoniae and SC via SpsA represents a novel biological interaction that might increase virulence by the impairment of bacterial clearance. PMID- 9350868 TI - Intracellular targeting of exoenzyme S of Pseudomonas aeruginosa via type III dependent translocation induces phagocytosis resistance, cytotoxicity and disruption of actin microfilaments. AB - Exoenzyme S (ExoS) is an ADP-ribosyltransferase secreted by the opportunistic pathogen Pseudomonas aeruginosa. The amino-terminal half of ExoS exhibits homology to the YopE cytotoxin of pathogenic Yersinia. Recently, YopE was found to be translocated into the host cell by a bacteria-cell contact-dependent mechanism involving the ysc-encoded type III secretion system. By using an approach in which exoS was expressed in different strains of Yersinia, including secretion and translocation mutants, we could demonstrate that ExoS was secreted and translocated into HeLa cells by a similar mechanism to that described previously for YopE. Similarly to YopE, the presence of ExoS in the host cell elicited a cytotoxic response, correlating with disruption of the actin microfilament structure. A similar cytotoxic response was also induced by a mutated form of ExoS with a more than 2000-fold reduced ADP-ribosyltransferase activity. However, the enzymatically active ExoS elicited a more definite rounding up of the HeLa cells, which also correlated with decreased viability of the cells after prolonged infection compared with cells infected with strains expressing mutated ExoS or YopE. This suggests that ExoS can act through two different mechanisms on the host cell. The expression of ExoS by Yersinia also mediated an anti-phagocytic effect on macrophages. In addition, we present evidence that extracellularly located P. aeruginosa is able to target ExoS into eukaryotic cells. Taken together, our data suggest that P. aeruginosa, by analogy with Yersinia, targets virulence proteins into the eukaryotic cytosol via a type III secretion-dependent mechanism as part of an anti-phagocytic strategy. PMID- 9350869 TI - Cascade regulation of the two CRP/FNR-related transcriptional regulators (ANR and DNR) and the denitrification enzymes in Pseudomonas aeruginosa. AB - ANR, an analogue of Escherichia coli FNR, has been shown to be necessary for denitrification, arginine deiminase activity and cyanide production of Pseudomonas aeruginosa. Another CRP/FNR-related regulator, DNR, has also been shown to be necessary for denitrification. In this study, we have found that the transcription of the dnr gene is under the control of ANR. When the dnr gene was expressed by tac promoter in an anr mutant, the strain recovered the ability to grow under anaerobic conditions by denitrification. Nitrate, nitrite, nitric oxide and nitrous oxide reducing activities were expressed, and the structural genes for nitrite and nitric oxide reductases were transcribed under anaerobic conditions in the anr mutant strain transformed with the dnr gene. These findings suggest that the expression of the denitrification system is controlled not directly by ANR but indirectly via DNR. The arginine deiminase activity and cyanide production were not regulated by DNR. PMID- 9350870 TI - Interspecies sequence differences in the Mip protein from the genus Legionella: implications for function and evolutionary relatedness. AB - The nucleotide sequence of the mip genes and their inferred amino acid sequences were determined from 35 Legionella species and compared with the published sequences for L. pneumophila, L. micdadei and L. longbeachae. The sequences were 69-97% conserved at the nucleotide level and 82-99% at the amino acid level, with total conservation of amino acids determined to be associated with sites known to be involved in peptidyl prolyl cis-trans isomerase activity. No apparent difference could be determined in the arrangement of amino acids that would predict a functional difference in Mip from species associated with disease and Mip from species isolated only from the environment. Additionally, a phylogenetic comparison of the sequences with published 16S RNA sequences, using both genetic distance and maximum parsimony methods, was performed. Few relationships were apparent that were well supported by both data sets, the most robust being a clade comprising ([(cincinnatiensis, longbeachae, sainthelensi, santicrucis) gratiana] (moravica, quateirensis, shakespearei, worsleiensis) anisa, bozemanii, cherrii, dumoffii, gormanii, jordanis, parisiensis, pneumophila, steigerwaltii, tucsonensis, and wadsworthii). PMID- 9350871 TI - Phage display selection of peptides against enzyme I of the phosphoenolpyruvate sugar phosphotransferase system (PTS). AB - The bacterial phosphoenolpyruvate-sugar phosphotransferase system (PTS) mediates the uptake and phosphorylation of carbohydrates and is involved in signal transduction. In response to the availability of carbohydrates it modulates catabolite repression, intermediate metabolism, gene expression and chemotaxis. It is ubiquitous in bacteria but does not occur in animals and plants. Uniqueness and pleiotropic function make the PTS a target for new antibacterial drugs. Enzyme I is the first component of the divergent protein phosphorylation cascade of the PTS. It transfers phosphoryl groups from phosphoenolpyruvate to the general phosphoryl carrier protein HPr. Six 15-mer, nine 10-mer and nine 6-mer peptides that inhibit enzyme I were selected from phage display libraries. Of these, 16 were synthesized and characterized. The majority of the peptides contain a histidine with an adjacent arginine. Two peptides were found to contain cysteines but no histidine. All peptides are rich in basic residues and lack acidic amino acids. The peptides inhibit the phosphotransferase system in vitro with IC50 of between 10 microM and 2 mM. Some, but not all, of the peptides inhibit cell growth in the agar diffusion test by an as yet undefined mechanism. All peptides are phosphorylated by enzyme I, and some are regenerated by slow autocatalytic hydrolysis of the phospho-peptide bond. PMID- 9350872 TI - Differences in the control of the temperature-dependent expression of four genes for desaturases in Synechocystis sp. PCC 6803. AB - Cyanobacteria are capable of desaturating the fatty acids in their membrane lipids in response to decreases in temperature. The cyanobacterium, Synechocystis sp. PCC 6803, contains four desaturases, which specifically catalyse desaturation at the delta6, delta9, delta12 and omega3 positions of fatty acids. The levels of the mRNAs transcribed from the genes that encode the delta6, delta12 and omega3 desaturases increased about 10-fold, but at different rates, upon a decrease in temperature from 34 degrees C to 22 degrees C, whereas the level of the mRNA for the delta9 desaturase remained constant. The increases in the levels of mRNAs were caused both by the enhanced transcription and by the increased stability of the mRNAs at the low temperature. Western blotting analysis demonstrated that levels of the delta6, delta12 and omega3 desaturases increased at different rates at the low temperature, while that of the delta9 desaturase remained constant. These observations indicate that the expression of the genes for the four desaturases is regulated by temperature in different ways. PMID- 9350873 TI - Systematic base composition variation around the genome of Mycoplasma genitalium, but not Mycoplasma pneumoniae. PMID- 9350874 TI - A tRNA(2Arg) gene of Corynebacterium diphtheriae is the chromosomal integration site for toxinogenic bacteriophages. PMID- 9350875 TI - A novel family of proteins that regulates antibiotic production in streptomycetes appears to contain an OmpR-like DNA-binding fold. PMID- 9350876 TI - Keeping a cool head, post-hypoxic hypothermia--an old idea revisited. AB - Hypoxia-ischaemia produces permanent brain damage by processes that continue for many hours after reoxygenation/reperfusion. This provides a window of opportunity for therapy aimed at preventing further loss of brain cells. Reducing brain temperature by 2-6 degrees C for 3-72 h after reoxygenation/reperfusion has been shown to reduce brain damage by 25-80% in controlled trials with six different neonatal animal models of hypoxia-ischaemia. No adverse effects from mild hypothermia have been documented. The mechanisms of protection are unknown but may include a reduction in extracellular excitotoxic amino acids, reduced nitric oxide synthesis and inhibition of apoptosis. Mild hypothermia is currently the most promising clinically feasible neural rescue therapy for full-term infants at risk of developing hypoxic-ischaemic encephalopathy, but clinical use must be restricted to approved trial protocols. PMID- 9350877 TI - The doula: an essential ingredient of childbirth rediscovered. AB - Eleven randomized control trials examined whether additional support by a trained lay person (called a doula), student midwife or midwife, who provides continuous support consisting of praise, encouragement, reassurance, comfort measures, physical contact and explanations about progress during labor, will affect obstetrical and neonatal outcomes. The women were healthy primigravidas at term. Meta-analysis of these studies showed a reduction in the duration of labor, the use of medications for pain relief, operative vaginal delivery, and in many studies a reduction in caesarian deliveries. At 6 weeks after delivery in one study a greater proportion of doula-supported women were breastfeeding, reported greater self-esteem, less depression, a higher regard for their babies and their ability to care for them compared to the control mothers. Observations during labor showed that fathers remained farther away from mothers than doulas, talked and touched less. When the doula was present with the couple during labor the father offered more personal support. The father-to-be' s presence during labor and delivery is important to the mother and father, but it is the presence of the doula that results in significant benefits in outcome. PMID- 9350878 TI - Ongoing challenges in pediatric hospice care. AB - Reports have begun to proliferate throughout the world which describe various models of pediatric hospice care. While encouraging, these reports identify universal obstacles that continue to compromise effective care. Challenges persist in areas of pain management, medical ethics, program administration, cost analysis, staff development and bereavement follow-up. Cooperative efforts are encouraged to address these issues. PMID- 9350879 TI - Effect of modest hypothermia on the immature kidney. PMID- 9350880 TI - Multiple food allergy: a possible diagnosis in breastfed infants. AB - Six infants suspected of food allergy during breastfeeding were evaluated using prick tests, total IgE, RASTs and intestinal permeability measurements during fast and provocation with mother's milk. An elimination diet was undertaken in mothers, removing first cow's milk protein (CMP), then, when inefficient, all foods suspected on the clinical history or a positive prick test in the child, followed by oral challenges in mother's diet with the corresponding food. The sole CMP-free diet in mothers always proved insufficient. In four, an additional diet excluding two to three other foods cleared the symptoms. Oral provocations in mother's diet with those foods were positive in all. In two, mothers turned down a diet excluding more than four foods, symptoms cleared while feeding the child with an extensively hydrolysed formula, whereas challenges with mother's milk induced immediate reactions. Intestinal permeability was altered during provocation tests with mother's milk sampled before maternal diet. Food allergy during breastfeeding may be due to multiple foods and the inefficacy of the sole CMP elimination in mothers does not rule out food sensitization. PMID- 9350881 TI - Oral rehydration with a plantain flour-based solution in children dehydrated by acute diarrhea: a clinical trial. AB - A clinical trial was conducted in order to prove the efficacy of a solution containing 50 g/l of plantain flour and 3.5 g/l of sodium chloride (NaCl) for the rehydration of children with acute diarrheal diseases. 121 children were given WHO-ORS (Group A) and 117 a plantain flour-based solution (Group B). Rehydration was successful in 85.9% in Group A and 88.0% in Group B (p = 0.634). Rehydration was completed in 5.28 h (SD 1.99) in Group A and in 4.88 h (SD 2.11) in Group B (p = 0.132). The average solution intake for rehydration was 24.56 ml/kg/h (SD 10.12) in Group A and 21.17 ml/kg/h (SD 9.35) in Group B (p = 0.00782). The mean stool output during rehydration was 8.45 g/kg/h (SD 9.72) in Group A and 4.69 g/kg/h (SD 4.98) in Group B (p = 0.00053). Decrease in blood levels of sodium and potassium occurred in some children in group B. The plantain flour-based solution proved effective for the treatment of dehydration due to acute diarrheal diseases and should be considered as an alternative when standard WHO-ORS is not available. PMID- 9350882 TI - High dose vitamin A supplementation in the course of pneumonia in Vietnamese children. AB - We carried out a randomized, placebo-controlled, double-blinded trial to evaluate the effect on morbidity of high dose oral vitamin A, given on hospital admission to 592 children aged 1-59 months with moderate and severe pneumonia. Severely underweight children were not included, but 45% were moderately underweight. The vitamin A and placebo groups were comparable in baseline characteristics. Four patients died. Among all of the surviving children, no differences were found regarding mean time for normalization of fever, respiratory rate and time of hospitalization. Stratification for moderate malnutrition, degree of pneumonia, age and sex revealed moderately malnourished vitamin A-supplemented children to have a shorter time of hospitalization (p = 0.04), due to an effect in females aged > 12 months (p = 0.02) and females with very severe pneumonia (p = 0.048). This study indicates that, in developing countries like Vietnam, supplementation with vitamin A in children with pneumonia could shorten the recovery rate in the ones that are undernourished, especially females > 1 y old. PMID- 9350883 TI - Salmonella meningitis: clinical experience of third-generation cephalosporins. AB - Fifteen paediatric patients with Salmonella meningitis were retrospectively reviewed. Presenting symptoms and signs included fever, vomiting, seizures, poor activity, diarrhoea and bulging anterior fontanelle in most patients. Seven out of eight patients with prolonged fever for > 10 days had neurologic sequelae; therefore, prolonged fever is a significant prognostic factor of a poor outcome (p < 0.005). All 15 patients had a brain ultrasound or computed tomography in the acute stage and 11 patients had abnormal findings. The 14 surviving patients were treated with a third-generation cephalosporin for at least 3 weeks. Seven patients (47%) made complete recoveries; two of them were treated solely with a third-generation cephalosporin. Only one mortality (6%) occurred and there were no relapses. In conclusion, high frequencies of prolonged fever, neuroimaging abnormalities and neurologic sequelae were seen in patients with Salmonella meningitis treated with third-generation cephalosporins. PMID- 9350884 TI - Interleukin 6, but not tumour necrosis factor-alpha, is a good predictor of severe infection in febrile neutropenic and non-neutropenic children with malignancy. AB - OBJECTIVE: Interleukin-6 (IL6), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) are important mediators of the inflammatory response in human infection. The aim of this study was to determine the relationship between serum levels of IL6, TNF-alpha, IFN-gamma and CRP in febrile children with malignant disease, and relate these levels to aetiology of fever, presence of neutropenia and the effect of untreated malignancy. METHODS: 110 febrile episodes in 70 children with malignant disease were included. Cytokine analyses were performed with sensitive immunoradiometric methods using double monoclonal antibodies. RESULTS: IL6 had a sensitivity of 74% in detecting sepsis in children with fever and malignant disease. This sensitivity was not influenced by the presence of neutropenia or newly diagnosed malignancy. A positive correlation between IL6 and the CRP levels on the following day was observed (r = .53). TNF alpha was elevated in 22% of the episodes and mean levels were significantly higher in untreated malignancy but lower in neutropenic patients. IFN-gamma was elevated in 18% of cases and correlated strongly with mean TNF-alpha levels. CONCLUSIONS: IL6 is a sensitive and early predictor of bacterial infection in both neutropenic and non-neutropenic febrile children with malignancy. It is more sensitive than CRP in detecting sepsis, but the predictive value is too low to allow IL6 levels to influence initial treatment decisions in patients with granulocytopenia. TNF-alpha production seems to be impaired in neutropenic children and serum TNF-alpha cannot be employed as an indicator of bacterial infection. PMID- 9350885 TI - The impact of thyroid autoimmunity in children and adolescents with Down syndrome. AB - The extent to which autoimmunity contributes to thyroid dysfunction in individuals with Down syndrome (DS) has not been clarified. In this study, we used the same highly sensitive method to detect both thyroid autoantibodies (thyroglobulin and thyroid peroxidase autoantibodies) in 70 children (32 M and 38 F) with DS, mean age 10.5 y (range 1-19 y). Twenty-seven (39%) of the patients were found to have thyroid autoantibodies, the prevalence of antibody positivity increasing with age. Of the 17 (24%) of the series who were hypothyroid (i.e. high basal TSH level and a low total- or free-T4 level), 11 had thyroid autoantibodies, and another 6 with thyroid autoantibodies became hypothyroid during 13-35 months of follow-up. Thus, the findings suggest that the majority of hypothyroid children with DS suffer from autoimmune thyroid disease. PMID- 9350886 TI - Constructional dyspraxia in preterm diplegia: isolation from visual and visual perceptual impairments. AB - OBJECTIVE: To evaluate ophthalmological profiles, visual perception and constructional function in preterm children with spastic diplegia (SD) and to clarify their neuropsychological deficits in comparison with a control group. METHODS: Thirty-five SD and 34 control children were investigated for visual acuity, eye position, stereoacuity, depth perception, visual perception, visuo spatial construction and constructional praxis. Each of the results was compared among the four groups as SD with and without strabismus, and control with and without strabismus. RESULTS: Strabismic SD showed worse visual acuity, worse stereoacuity and worse depth perception than the other groups. Constructional dyspraxia was detected in 94.1% of SD either with or without strabismus, while it was rare in the control group. There was no significant contribution of visual acuity, eye position, stereoacuity or depth perception to constructional dyspraxia by stepwise multiple linear regression analysis. CONCLUSION: Strabismic preterm SD children are at high risk for visual dysfunction. Constructional dyspraxia was frequently found in SD children and may be a dysfunction isolated from ophthalmological and visual perceptual dysfunctions. PMID- 9350887 TI - CNS tumours in south-western Finland: high location, high incidence. AB - The Department of Paediatrics at the University Central Hospital of Turku, Finland has 130,000 children under 17 y of age within its catchment area. We collected all 103 cases of newly diagnosed CNS tumours from the 15-y period of 1981-95. The incidence was 5.3:100,000, a figure twofold those usually presented. During the period 1981-85 the incidence was lower (4) than during the subsequent 5-y periods (5.7 and 6.2). There were no statistical differences between the incidences of the supra- vs infratentorial brain tumours. Optic glioma was unusually common (17%, CI 13.9-20%). PMID- 9350888 TI - Immune deficiencies in chronic intestinal pseudo-obstruction. AB - AIM: Chronic intestinal pseudo-obstruction has been associated with urinary disorders, myopathy, and ophthalmoplegia in adults and cholelithiasis in children. We observed a high percentage of total-parenteral-nutrition-dependent patients with pseudo-obstruction and recurrent infections requiring gammaglobulin infusions. METHODS: All records for 23 children with chronic intestinal pseudo obstruction (10 females and 13 males, mean age 9.8 y +/- 4.9 y, range 4-24 y) referred for a nutritional evaluation from 1992 to 1995 were reviewed. Chronic intestinal pseudo-obstruction was diagnosed by clinical, radiographic findings and antroduodenal manometry. Intestinal full-thickness biopsies were performed in seven children. RESULTS: Hypogammaglobulinemia was diagnosed in 18 patients (78%): 16 patients had various immunoglobulin deficiencies and 2 had selective antibody deficiency. Intravenous gammaglobulin was administered in 14 patients. Other medical conditions affecting the children are summarized as follows: autonomic dysfunction in 10 patients (43%), recurrent hypoglycemia in 9 (39%), asthma in 9 (39%), cholecystitis in 7 (30%), low serum carnitine level in 6 (26%), urinary dysfunction in 6 (26%), pancreatitis in 5 (22%), behavioral problems in 5 (22%), myopathy in 2 (9%), idiopathic thrombocytopenia in 2 (8%), velopharyngeal insufficiency in 1 (4%), oculocutaneous albinism in 1 (4%), Pierre Robin syndrome in 1 (4%), and protein C deficiency in 1 (4%). Munchausen syndrome was suspected in two patients. CONCLUSIONS: Chronic intestinal pseudo-obstruction appears to be associated with immune deficiencies. It is unclear if the immune deficiencies, intestinal pseudo-obstruction, and the other medical conditions have a common underlying etiology. Repeated infections may be due to impaired immune function in children with chronic intestinal pseudo-obstruction. We recommend screening for immune deficiencies in children with chronic intestinal pseudo-obstruction. PMID- 9350889 TI - Glucose metabolism and insulin secretion in children with cyanotic congenital heart disease. AB - The aim of the study was to reveal differences in carbohydrate metabolism in children with cyanotic congenital heart diseases (CHD). Thirteen children with diseases of these kinds were investigated with regard to glucose tolerance and insulin secretion and comparisons were made with healthy controls of the same age. Investigations included an intravenous glucose tolerance test, insulin response to the glucose load in plasma and insulin secretion rate. The results reveal lower fasting glucose levels and signs of a higher insulin secretion rate in the relatively few patients in the CHD group where C-peptide measurements were performed, but no differences in glucose tolerance. The reasons for the differences are unclear, but the chronic increases in circulating catecholamines in combination with the impaired nutritional status of these children with CHD are probably the most important factors. We conclude that these divergences in carbohydrate metabolism should be emphasized in the care of children with CHD. PMID- 9350890 TI - Evaporation rate and skin blood flow in full term infants nursed in a warm environment before and after feeding cold water. AB - Earlier results have shown that some infants born by elective Caesarean section start to sweat in a warm environment while others do not, and that sweating can be inhibited by feeding cold glucose. To determine whether these earlier observations, indicating a difference in postnatal temperature adaptation, could be reproduced in vaginally born infants, we measured the rate of evaporation from the skin surface, body and skin temperatures from several sites, skin blood flow and respiratory rate in newborn infants nursed in a warm environment, before and after feeding cold water. In all infants the body and skin temperatures increased in the warm environment (p < 0.01), with a decreasing difference between oesophageal and leg skin temperature (p < 0.01). Visible sweating occurred in 9/14 infants at a rectal temperature of 37.5 degrees C. In the infants who started to sweat, evaporation rate increased from 5.6 +/- 2.8 (SD) g/m2/h 15 min before sweating to 15.7 +/- 10.6 g/m2/h (p < 0.05) when sweat became visible and the infants were fed cold water. After feeding of cold water the evaporation rate decreased and within 10 min returned to a value not significantly different from the pre-sweating value. Interscapular skin blood flow had increased by 42% (p < 0.01) at the time of sweating and decreased by 22% (p < 0.01) after feeding cold water. In the infants who did not start to sweat, no increase in evaporation rate was noted and the changes in skin blood flow were not statistically significant. The infants who started to sweat did not differ from those who did not regarding maternal medication during delivery. We conclude that some, but not all, newborn infants start to sweat at a body temperature of 37.5 degrees C. In the infants who start to sweat, sweating and an increase in skin blood flow can be inhibited by feeding cold water. There seem to be individual differences in the regulation of body temperature in newborn infants, possibly due to a delayed change in the central temperature set-point in some infants. PMID- 9350891 TI - Respiratory distress syndrome in infants with impaired intrauterine growth. AB - The recently introduced intrauterine growth curve, based on ultrasonically estimated foetal weights, was retrospectively applied to an inborn population of 883 infants born before 33 gestational weeks at the University Hospital of Lund, during 1985-94. The estimation of birthweight deviation resulted in 630 (71.3%) infants with a birthweight appropriate for gestational age (AGA), 244 (27.6%) infants with a birthweight small for gestational age (SGA) and 9 (1.1%) infants with a birthweight large for gestational age. Birthweight deviation was associated with an increased mortality [odds ratio (OR) adjusted for gestational age 1.29 per SD (12%) change in birthweight for gestational age, 95% CI: 1.10 1.50; p = 0.002]. At gestational age 25-28 weeks, SGA-infants had an increased incidence of respiratory distress syndrome (RDS) as compared to AGA-infants (OR adjusted for gestational age: 1.98, 95% CI: 1.12-3.52; p = 0.019). At gestational age 29-32 weeks, SGA-infants had a lower incidence of RDS as compared to AGA infants (OR adjusted for gestational age: OR 0.52, 95% CI: 0.34-0.80; p = 0.003). After adjustment for confounding variables, infants born at gestational age 25-28 weeks from mothers with pre-eclampsia, appeared to be a high-risk group for RDS, whereas at the age of 29-32 gestational weeks, negative birthweight deviation had a protective effect against RDS. Antenatal corticosteroid administration appeared to have a less beneficial effect on mortality, RDS and cerebral haemorrhage in infants born SGA vs in those born AGA. PMID- 9350892 TI - Rapid detection of neonatal sepsis using polymerase chain reaction. AB - Clinical diagnosis of sepsis in newborn infants is not easy and there is no laboratory test with 100% specificity and sensitivity, with the exception of blood culture, the results of which are not available for at least 48-72 h. Polymerase chain reaction methodology has been used to diagnose different bacterial, viral and protozoal infections, and the possibility of amplifying the DNA region common to all bacteria could represent an optimal method for the diagnosis of sepsis. The authors have performed PCR in a group of 33 neonates at risk for early-onset sepsis, correlating molecular data with blood culture results. The presence of bacterial DNA in blood samples was evaluated, amplifying the DNA region encoding the 16S rRNA. There were no false negative results (four positive blood cultures and four positive PCR), with competitive costs and time. This method also allows the diagnosis of sepsis due to uncommon species and also, using a second PCR with specific primers, an aetiological diagnosis. PMID- 9350893 TI - Coagulation and fibrinolytic systems in the ill preterm newborn. AB - AIM: The activation pattern of the clotting and fibrinolytic systems in 63 preterm infants (GA 31, 6 +/- 2.3 weeks) was studied. METHODS: The infants were divided into four groups: (i) IRDS, (ii) asphyxia at birth, (iii) sepsis, and (iv) mild infection. A control group was composed of preterm infants without any apparent disease (GA 32 +/- 1.8 weeks). RESULTS: During IRDS we found a systemic activation of both coagulation and fibrinolysis at birth which was represented by lower levels of ATIII (27.7 +/- 8.8%) and significantly greater levels of TAT (37.9 +/- 31.9 ng/ml), D-dimers (1242.7 +/- 206.9 ng/ml), tPA Ag (10.9 +/- 5.3 ng/ml) and PAI Ag (59.9 +/- 16.7 ng/ml) than in the control group. In the asphyxiated newborns there were no significant differences from the controls. During their seventh day of life, a significant reduction of all the analysed parameters (TAT, D-dimers, tPA, PAI) and a significant increase in ATIII were seen in the newborns with IRDS, while no significant modification was observed in the newborns with asphyxia at birth. When the newborns with sepsis were compared with those with mild infection, their TAT and PAI values proved to be significantly higher for the first tests (21.7 +/- 18.8 vs 9.2 +/- 6.9 microg/l and 53.6 +/- 14.4 vs 37.7 +/- 10.2 ng/ml respectively). During the second tests, 7 days later, only TAT (16.7 +/- 14.7 vs 6.3 +/- 4 microg/l) levels remained high while D-dimers (1094.2 +/- 400.6 vs 646 +/- 200ng/ml) and tPA (11.3 +/- 8 vs 4.9 +/- 2 ng/ml) were significantly higher in the septic group of newborns than those with mild infection. CONCLUSIONS: These data indicate that there is an activation of the clotting and fibrinolytic systems both in the initial phase of IRDS as well as during sepsis. PMID- 9350894 TI - Plasma thrombomodulin level in very low birthweight infants at birth. AB - OBJECTIVE: Plasma soluble thrombomodulin level reflects endothelial damage. The plasma thrombomodulin level at birth is increased in asphyxiated full-term infants. There is no report of plasma thrombomodulin level in premature infants. To determine the thrombomodulin level in premature infants and whether it might reflect endothelial damage, we examined the plasma thrombomodulin level in very low birthweight (VLBW) infants at birth. METHODS: Forty-five VLBW infants, of whom 14 had perinatal asphyxia complications, were recruited. As a control, 50 full-term infants without complications were also studied. Plasma thrombomodulin concentration, pH, base deficit, serum creatinine and D-dimer concentration, platelet count and fibrinogen concentration were measured within 1 hour after birth. RESULTS: There were significant differences in plasma pH, creatinine concentration, platelet count, antithrombin III activity and D-dimer concentration between VLBW infants and full-term infants. Plasma thrombomodulin concentration (39.0 (16.6-93.7) vs 27.0 (16.6-39.1) microg/L, p < 0.0001) and plasma thrombomodulin-to-serum creatinine ratio (0.82 (0.19-2.65) vs 0.47 (0.24 0.70) microg/micromol, p < 0.0001) were significantly higher in VLBW infants than those in full-term infants. By univariate analyses for all neonates, there were significant relations between plasma thrombomodulin concentration and gestational age, birthweight, plasma pH, creatinine concentration, platelet count and antithrombin III activity. A stepwise multiple linear regression model using the above variables as dependent factors showed only birthweight contributed significantly to plasma thrombomodulin concentration (plasma thrombomodulin concentration (microg/l) = 45.677-0.006 (birthweight; g), r2 = 0.323, p < 0.0001, n = 94). Plasma thrombomodulin concentration and plasma thrombomodulin-to -serum creatinine ratio in VLBW infants with asphyxia were higher than in those without asphyxia, but not significantly different (43.2 +/- 17.7 vs 38.3 +/- 8.5 microg/l and 0.92 +/- 0.60 vs 0.83 +/- 0.37 microg/micromol). CONCLUSION: Plasma thrombomodulin level in VLBW infants shows a high value at birth, and we consider the main factor responsible for this elevation may be endothelial damage or low clearance rate of thrombomodulin, which may be related to early gestational age. PMID- 9350895 TI - Prevalence of asthma in Danish children aged 8-10 years. AB - The aim of the study was to estimate the point prevalence of asthma in schoolchildren aged 8-10 y in the County of Copenhagen, Denmark. In all, 1040 schoolchildren were randomly chosen for the study and a total of 774 completed the study. The suspicion of asthma was based on a questionnaire about respiratory symptoms and on daily registration of respiratory symptoms and peak expiratory flow rate (PEF) for 4 weeks. The conclusive diagnosis was based on interview, clinical examination, spirometry and an exercise test. The prevalence of children with asthma diagnosed by their GPs was 31/774 (4.0%). A further 20/774 (2.6%) were diagnosed as having asthma. There is evidence to suggest that asthma is less frequent in Denmark than in Great Britain, New Zealand and Australia. PMID- 9350896 TI - How do they grow? A study of south-eastern Nigerian adolescent girls. AB - This study assessed the nutritional status of Nigerian adolescent girls living in two areas of south-eastern Nigeria. A cross sectional survey was undertaken in a rural village in Ogoniland, and five secondary schools in Port Harcourt, south eastern Nigeria. All (386) menarcheal girls aged 14-19 y living in the rural village, and a stratified cluster sample (845) of menarcheal girls aged 14-19 in the five urban schools were investigated. Mean heights and weights of rural girls were around -1 Z-score below the British reference median. 10.4% of rural and 4.7% of urban girls were stunted (< OR =2nd centile, British 1990 reference values). After calculating mean body mass index-for-age, various cut-off points for low body mass index were tested. At a cut-off of < OR =9th centile, 15.6% of rural and 8.0% of urban girls would be classified as thin. Girls with a haemoglobin <10.Og/dl were significantly more likely to have a low body mass index than those with haemoglobin values > OR =10.0 g/dl. More studies are needed to refine the definition and interpretation of low body mass index in adolescents. PMID- 9350897 TI - Secular trend in the sexual maturation of southern Chinese girls. AB - In 1993, a cross-sectional study of sexual maturation of normal Chinese schoolgirls was performed in Hong Kong. The aim of the study was to obtain an up to-date reference for normal pubertal development in Chinese girls. Breast development was assessed in 3749 girls aged 7-19 y, and pubic hair rating was assessed in 3745 girls. Menstrual status was recorded in 6467 girls over 6 y of age. The median age of onset of puberty as indicated by breast stage II or above was 9.78 (95% CI 9.70-9.85) y. The median age of onset of pubic hair development was 11.64 (95% CI 11.56-11.72) y. The median age of menarche was 12.38 (95% CI 11.98-12.78) years. Percentile values for the age at which each puberty staging appeared were constructed and incorporated into the height-for-age charts. When comparison is made with similar studies done in 1962 and 1979, a significant downward secular trend in sexual maturation is observed (p < 0.01). Except for breast development the downward secular trend in sexual maturation appears to be diminishing and may be coming to a halt in the Chinese girls in Hong Kong. Their median ages of sexual maturation are now among one of the earliest medians recorded in the world population studied. PMID- 9350898 TI - Terminal care of the child with cancer at home. AB - One hundred paediatric patients with either leukaemia (36%), solid tumours (34%) or brain tumours (30%), treated at the Children's Hospital, University of Helsinki, Finland, died during 1987-92; 70 of them died while in organized terminal care. They were treated at home (60%), in hospital (29%), and partly at both (11%). One or both parents stayed at home to take care of their child. Personnel of the oncologic ward coordinated home care. The purpose of this study was to evaluate the advantages and disadvantages of a terminal care program, with special reference to terminal care at home. Evaluation included retrospective analysis of patients' records, as well as a structured interview with the two parents separately. The quality of life of the children during the terminal period was greatly influenced by their happiness at being at home. Relief of symptoms, particularly pain, was in most instances adequate. Most parents had no complaints to make afterwards. Only some of them complained of having received too little information, too little supervision and support, and insufficient preparation for the death of the child. Thus, the system of terminal care at home proved satisfactory for the child and the whole family in many different respects. For successful home care, the parents need continuous supervision, help and support by well-trained personnel. PMID- 9350899 TI - Bedwetting and behavioural and/or emotional problems. AB - OBJECTIVE: To assess the link between enuresis nocturna and the severity of behavioural and/or emotional problems in Dutch children and the course of these problems. SETTING: West-Mine Region in the Netherlands. SUBJECTS AND METHODS: Prospective cohort study involving 66 of the 80 bedwetting children from all 1652 children born in 1983 in this region. After 1 y, contact was still possible with 64 of the enuretics. We used the Dutch version of the Child Behaviour Checklist (CBCL) and a questionnaire about bedwetting. RESULTS: The mean T-score for Total Problems (CBCL score) in 1992 (M1; mean age 8.6) was 52.1, and 1 y later was 49.2 (M2). There was no significant difference in the CBCL scores for M1, M2 and a matching group from the Dutch CBCL norm population, either in the group who remained wet or in the group who became dry. There were no differences between the sexes. There was no link between the severity of behavioural and emotional problems and the frequency of bedwetting. However, more children with bedwetting than expected were in the clinical range. CONCLUSION: There was no difference in behavioural and/or emotional problems between the first and the second measurement and the matching group from the CBCL norm group. There were no differences in behavioural and/or emotional problems between primary and secondary bedwetters, nor were there any consequences related to the frequency of bedwetting. PMID- 9350900 TI - Hepatitis B and C virus infections in Turkish children with haemophilia. AB - We examined 41 Turkish children with haemophilia for evidence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections using the enzyme-linked immunosorbent assay (ELISA). Hepatitis B surface antigen was found to be positive in 11 patients (26.8%) and HCV-specific antibody (anti-HCV) was detected in 10 (24.4%) patients. There was a close relationship of the number of transfusions of blood plasma to the presence of HCV specific antibody, but not to the serum markers of HBV infection. In countries where HBV infection is commonly seen and problems in transfusion practice continue, as in Turkey, children with haemophilia are at greater risk for HBV and HCV infections. PMID- 9350901 TI - Continuous mannose infusion in carbohydrate-deficient glycoprotein syndrome type I. AB - The effects on isoelectrofocusing patterns of serum glycoproteins were studied in a patient with CDG syndrome type I and phosphomannomutase deficiency during 3 weeks of continuous intravenous mannose infusion. Doses of 5.7 g/kg/day led to stable serum mannose levels up to 2.0 mmol/l and were well tolerated without signs of liver or renal toxicity. While most of the pathological glycoprotein patterns, including alpha1-antitrypsin, typical for CDG syndrome type I remained unchanged, mannose infusion led to a unique change of the isoelectrofocusing pattern of serum sialotransferrins with appearance of two extra bands after 3 weeks of treatment. PMID- 9350902 TI - Non-organic failure to thrive complicated by benign intracranial hypertension during catch-up growth. AB - Severe non-organic failure to thrive associated with physical and emotional abuse including food deprivation was diagnosed in a 9-y-old boy. Rapid catch-up growth (weight and height) followed change of carer. Recovery of poor growth hormone response to clonidine stimulation was associated with benign intracranial hypertension accompanied by headaches and vomiting. Possible mechanisms are discussed. PMID- 9350903 TI - Early clinical manifestation of glutaric aciduria type I and nephrotic syndrome during the first months of life. AB - We describe a male patient with glutaric aciduria type I which had already presented during the neonatal period with therapy-resistant seizures. In the course of the disease, he also developed choreoathetosis and dystonia. The disease was associated with nephrotic syndrome. Renal histology showed signs of a glomerular disorder with shrinking of glomerular tufts, increase in mesangial matrix, proliferation of extracapillary epithelial cells and formation of larger epithelial crescents. The child died at 22 weeks of age due to end-stage renal failure. This report illustrates an unusual and early clinical manifestation of glutaric aciduria type I and a hitherto unknown association with nephrotic syndrome in early childhood. PMID- 9350904 TI - We need a new understanding of the reabsorption of cerebrospinal fluid--II. PMID- 9350905 TI - Probiotics and prebiotics in the treatment of infections due to vancomycin dependent Enterococcus faecalis and of imbalance of the intestinal ecosystem (dysbiosis) PMID- 9350906 TI - Another cause of hyponatraemia in patients with bacterial meningitis: cerebral salt wasting. PMID- 9350908 TI - Regional remodeling of atherosclerotic arteries: a major determinant of clinical manifestations of disease. AB - In this review we present the current data on remodeling, based on in vivo ultrasound imaging or postmortem histologic analysis of native peripheral and coronary arteries from animal models and studies in patients (coronary artery saphenous vein bypass grafts, lesions of restenosis after balloon angioplasty and other catheter-based interventions). Histologic and ultrasound imaging studies of arteries with atherosclerosis and after vascular injury reveal that arterial remodeling is common and that the cross-sectional area of the vessel is not constant. Compensatory enlargement, inadequate compensatory enlargement and shrinkage at the site of atherosclerotic lesions occurs in coronary and peripheral arteries. Current studies demonstrate that arterial remodeling is a major determinant of vessel lumen size. PMID- 9350909 TI - Management strategies and determinants of outcome in acute major pulmonary embolism: results of a multicenter registry. AB - OBJECTIVES: The present study investigated current management strategies as well as the clinical course of acute major pulmonary embolism. BACKGROUND: The clinical outcome of patients with acute pulmonary embolism who present with overt or impending right heart failure has not yet been adequately elucidated. METHODS: The 204 participating centers enrolled a total of 1,001 consecutive patients. The inclusion criteria were based on the clinical findings at presentation and the results of electrocardiographic, echocardiographic, nuclear imaging and cardiac catheterization studies. RESULTS: Echocardiography was the most frequently performed diagnostic procedure (74%). Lung scan or pulmonary angiography were performed in 79% of clinically stable patients but much less frequently in those with circulatory collapse at presentation (32%, p < 0.001). Thrombolytic agents were given to 478 patients (48%), often despite the presence of contraindications (193 [40%] of 478). The frequency of initial thrombolysis was significantly higher in clinically unstable than in normotensive patients (57% vs. 22%, p < 0.001). Overall in-hospital mortality rate ranged from 8.1% in the group of stable patients to 25% in those presenting with cardiogenic shock and to 65% in patients necessitating cardiopulmonary resuscitation. Major bleeding was reported in 92 patients (9.2%), but cerebral bleeding was uncommon (0.5%). Finally, recurrent pulmonary embolism occurred in 172 patients (17%). CONCLUSIONS: Current management strategies of acute major pulmonary embolism are largely dependent on the degree of hemodynamic instability at presentation. In the presence of severe hemodynamic compromise, physicians often rely on the findings of bedside echocardiography and proceed to thrombolytic treatment without seeking further diagnostic certainty in nuclear imaging or angiographic studies. PMID- 9350910 TI - Pulmonary embolism for cardiologists. PMID- 9350911 TI - Population-based analysis of the effect of the Northridge Earthquake on cardiac death in Los Angeles County, California. AB - OBJECTIVES: We sought to determine whether a natural disaster affected total cardiovascular mortality and coronary mortality in an entire population. BACKGROUND: The effect of the January 17, 1994 Northridge Earthquake (NEQ) on all deaths and causes of deaths within the entire population of Los Angeles County is unknown. The purposes of our study were to analyze all deaths in this entire population before, during and after the NEQ and to determine whether the NEQ temporally and spatially altered death due to cardiovascular disease. METHODS: We analyzed all death certificate data (n = 19,617) from Los Angeles County during January of 1992, 1993 (control periods) and 1994, using International Classification of Diseases, 9th Revision codes for ischemic heart disease (IHD) and atherosclerotic cardiovascular disease (ASCVD), as well as other causes of death. RESULTS: There was an average of 73 deaths per day due to IHD and ASCVD during January 1 to 16, 1994; this increased to 125 on the day of the NEQ, and then decreased to 57 deaths per day from January 18 to 31 (p < 0.00001, before NEQ vs. day of NEQ; after NEQ vs. day of NEQ; and before NEQ vs. after NEQ). The NEQ was associated with an increase in deaths due to myocardial infarction and trauma but not cardiomyopathy, hypertensive heart disease, valvular heart disease, cerebrovascular disease or noncardiovascular causes. Based on plots of daily deaths due to IHD and ASCVD, the decrease in deaths during the 14 days after the NEQ (-144) overcompensated for the increase on the day of the NEQ (+55). Geographic analysis revealed a redistribution of deaths due to IHD and ASCVD toward the epicenter on the day of the NEQ. CONCLUSIONS: When an entire population simultaneously experiences a major environmental stress, there is an increase in death due to coronary artery disease (but not other cardiac causes), followed by a decrease that overcompensates for the excess of death. The overcompensation may represent a residual population that is more resistant to stress or a possible preconditioning effect of the stress, or both. This study supports the concept that cardiovascular events within an entire population can be triggered by a shared stress. PMID- 9350912 TI - Quantification of the benefit of earlier thrombolytic therapy: five-year results of the Grampian Region Early Anistreplase Trial (GREAT). AB - OBJECTIVES: This report presents the 5-year results of the Grampian Region Early Anistreplase Trial (GREAT) and quantifies the benefit of earlier thrombolysis in terms that are generally applicable. BACKGROUND: Although it is accepted that the earlier thrombolytic therapy is given for acute myocardial infarction the greater the benefit, there are widely differing estimates of the magnitude of the time related benefit of thrombolysis because of inappropriate trial design and analysis. METHODS: In a previously reported randomized trial, anistreplase (30 U) was given intravenously either before hospital admission or in the hospital, at a median time of 105 and 240 min, respectively, after onset of symptoms. Intention to treat and multivariate analyses of the 5-year results were performed. RESULTS: By 5 years, 41 (25%) of 163 patients had died in the prehospital treatment group compared with 53 (36%) of 148 in the hospital treatment group (log-rank test, p < 0.025). Delaying thrombolytic treatment by 1 h increases the hazard ratio of death by 20%, equivalent to the loss of 43/1,000 lives within the next 5 years (95% confidence interval 7 to 88, p = 0.012). Delaying thrombolytic treatment by 30 min reduces the average expectation of life by approximately 1 year. CONCLUSIONS: The magnitude of the benefit from earlier thrombolysis is such that giving thrombolytic therapy to patients with acute myocardial infarction should be accorded the same degree of urgency as treatment of cardiac arrest. Policies should be developed for giving thrombolytic therapy on-site if practicable and by the first qualified person to see the patient. PMID- 9350913 TI - Six-month outcome in patients with myocardial infarction initially admitted to tertiary and nontertiary hospitals. RESCATE Investigators. Recursos Empleados en el Sindrome Coronario Agudo y Tiempos de Espera. AB - OBJECTIVES: The aim of the present study was to ascertain whether the degree of accessibility to coronary angiography and revascularization results in differing usages or outcomes, or both, in the setting of a high coverage national health system. BACKGROUND: The selective use of coronary angiography and revascularization procedures in the management of acute myocardial infarction (MI) remains controversial. METHODS: A cohort of 1,460 consecutive patients with a first MI admitted to four referral teaching hospitals (one with tertiary facilities) were followed up for 6 months after admission. Only patients initially admitted to each of the study hospitals were retained for analysis in the original hospital's cohort. End points were 6-month mortality and readmission for reinfarction, unstable angina, heart failure or severe ventricular arrhythmia. RESULTS: Patients admitted to the tertiary hospital were more likely to undergo coronary angiography (adjusted relative risk 4.22, 95% confidence interval [CI] 3.37 to 5.45) than those admitted to the nontertiary sites (use rate: 22.1% for nontertiary care, 55.5% for tertiary care). Revascularization procedures were performed in 21.2% of patients in the tertiary hospital and in 8.3% in the nontertiary hospitals (p < 0.0001). Median delay for emergency coronary angiography was shorter in the tertiary hospital (within 1 vs. 2 days, p < 0.0001). Six-month mortality or readmission rates were similar (23.7% and 24.7% for tertiary and nontertiary care, respectively). After adjustment for comorbidity and disease severity, the relative risk of death or readmission for the tertiary hospital was 1.03 (95% CI 0.69 to 1.53) times that of the nontertiary hospitals. CONCLUSIONS: Selective use of coronary angiography and revascularization procedures may be as effective as less restricted use in the management of acute MI. PMID- 9350914 TI - Beneficial effect of intracoronary verapamil on microvascular and myocardial salvage in patients with acute myocardial infarction. AB - OBJECTIVES: We assessed the acute effect of intracoronary injection of verapamil on microvascular function after primary percutaneous translumanal coronary angioplasty (PTCA) for acute myocardial infarction (AMI) with myocardial contrast echocardiography (MCE) in relation to functional outcomes. BACKGROUND: Recent clinical studies have documented the potential of verapamil for possible increase in coronary blood flow after primary PTCA. METHODS: Forty patients with a first AMI were randomly assigned to the verapamil group (n = 20) or the control group (n = 20). In the verapamil group, verapamil (0.5 mg) was injected into the infarct-related artery shortly after PTCA, followed by the oral administration. We performed MCE with an intracoronary injection of sonicated microbubbles before and after verapamil. To assess microvascular integrity, we determined the baseline-subtracted peak intensity in the risk area and the ratio of the no reflow zone plus the low reflow zone to the risk area (low reflow ratio). We determined the average wall motion score (dyskinesia/akinesia = 3; normal = 0) in the risk area on the day of AMI and a mean of 24 days later. RESULTS: The low reflow zone was observed shortly after PTCA in 14 verapamil group patients, and the low reflow ratio decreased after verapamil (0.39 +/- 0.23 vs. 0.29 +/- 0.17 [mean +/- SD], p < 0.05). Peak intensity significantly (p < 0.05) increased from 6 +/- 5 to 12 +/- 6 after verapamil. The reduction in wall motion score from the acute (day -1) to the late stage (day -24) was significantly greater in the verapamil group than in the control group (0.7 +/- 0.8 vs. 0.2 +/- 1.3, respectively, p < 0.05). CONCLUSIONS: Intracoronary administration of verapamil after primary PTCA can attenuate microvascular dysfunction and thereby augment myocardial blood flow in patients with AMI, leading to better functional outcome than with PTCA alone. PMID- 9350915 TI - Mortality from coronary heart disease and cardiovascular disease among adult U.S. Hispanics: findings from the National Health Interview Survey (1986 to 1994). AB - OBJECTIVES: We sought to estimate the coronary heart disease (CHD) and cardiovascular disease (CVD) mortality experience of U.S. Hispanics. BACKGROUND: Limited information is available concerning the mortality from CHD among U.S. Hispanics, the nation's second largest minority group. METHODS: The study used data from the National Health Interview Survey (1986 to 1994), including representative national samples of 246,239 non-Hispanic whites, 38,042 blacks and 14,965 Hispanics who were > or = 45 years old at baseline. Mean follow-up of mortality was 5 years (range 1 to 10). RESULTS: During the follow-up period, 27,702 whites (11%), 4,976 blacks (13%) and 1,061 Hispanics (7%) died. Among men, the age-adjusted total mortality per 100,000 person-years was 3,089 in whites and 2,466 in Hispanics, and among women, it was 1,897 and 1,581 in whites and Hispanics, respectively. The Hispanic/white mortality rate ratio for CHD was 0.77 (95% confidence interval [CI] 0.64 to 0.93) and 0.82 (95% CI 0.66 to 1.01) for men and women, respectively. The rate ratio was 0.79 (95% CI 0.68 to 0.91) and 0.80 (95% CI 0.69 to 0.94), respectively, for mortality from cardiovascular diseases. Given the lower all-cause mortality in Hispanics, the proportion of total deaths due to CHD and CVD was similar between the two populations for the same gender and were, respectively, 29.7% and 44.7% in white men, 28.1% and 44.3% in Hispanic men, 24.9% and 43.2% in white women and 24.1% and 41% in Hispanic women. CONCLUSIONS: These data from a cohort of a large national sample are consistent with vital statistics that show that all-cause, CHD and CVD mortality is approximately 20% lower among adult Hispanics than among whites in the United States. PMID- 9350916 TI - A mutation in the methylenetetrahydrofolate reductase gene is not associated with increased risk for coronary artery disease or myocardial infarction. AB - OBJECTIVES: We sought to determine whether the C677T transition in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with increased risk for coronary artery disease (CAD) or myocardial infarction (MI). BACKGROUND: Elevated plasma homocysteine has been identified as a risk factor for coronary atherosclerosis. Homocysteinemia may result from deficient MTHFR activity. A thermolabile form of MTHFR, associated with a C677T genetic transition, shows reduced activity and may be a risk factor for CAD. METHODS: Blood was withdrawn from patients undergoing coronary angiography, and DNA was extracted by a phenol chloroform method. Genotyping was done by polymerase chain reaction (PCR) amplification of a 198-base pair segment of the MTHFR gene that brackets nucleotide 677. The amplicon was digested with the HinfI restriction enzyme. Products were visualized after electrophoresis in 1.5% agarose with ethidium bromide. RESULTS: Among 200 patients with a diagnosis of MI, the polymorphic allelic frequency was 33.3%, compared with 32.1% among 554 control subjects (p = 0.68); homozygosity was present in 11.5% of patients and 10.6% of control subjects (p = 0.74, odds ratio [OR] 1.09, 95% confidence interval [CI] 0.63 to 1.82). Among 510 patients with severe CAD (>60% stenosis), allelic frequency was 32.0%, compared with 34.8% for 168 subjects without CAD (<10% stenosis, p = 0.33); 11.2% of patients with CAD compared with 13.1% of control subjects were homozygous (p = 0.50, OR 0.83, 95% CI 0.5 to 1.40). CONCLUSIONS: Patients with angiographic evidence of CAD or clinical MI do not show an increased frequency of the C677T transition in the MTHFR gene. Our findings do not support this polymorphism as a risk factor for CAD or MI in a predominantly white, well nourished population of unrestricted age. PMID- 9350917 TI - HMG-CoA reductase inhibitors decrease CD11b expression and CD11b-dependent adhesion of monocytes to endothelium and reduce increased adhesiveness of monocytes isolated from patients with hypercholesterolemia. AB - OBJECTIVES: This study sought to determine whether inhibitors of 3-hydroxy-3 methyl-glutaryl coenzyme A (HMG-CoA) reductase affect CD11b expression and adhesiveness of monocytes in vitro and after treatment of patients with hypercholesterolemia. BACKGROUND: HMG-CoA reductase inhibitors improve survival of patients with coronary heart disease (CHD) and prevent CHD in hypercholesterolemic men. Because these drugs have been shown to modulate monocyte functions, they may act by reducing monocyte adhesion to endothelium, which is crucial in atherogenesis. METHODS: Isolated human blood monocytes were subjected to flow cytometric detection of CD11b and adhesion assays on fixed human endothelial cells after treatment with lovastatin in vitro or ex vivo before and after treatment of hypercholesterolemic patients with HMG-CoA reductase inhibitors. RESULTS: The integrin heterodimer CD11b/CD18 expressed on monocytes interacts with intercellular adhesion molecule-1 on endothelium and is involved in monocyte adhesion to endothelium. Treatment of monocytes with lovastatin in vitro slightly and dose dependently reduced surface expression of CD11b on monocytes. Moreover, lovastatin inhibited CD11b-dependent adhesiveness to fixed endothelium of unstimulated monocytes or monocytes stimulated with monocyte chemotactic protein 1. Coincubation with mevalonate, but not with low density lipoprotein (LDL), reversed the effects of lovastatin, suggesting that early cholesterol precursors, but not cholesterol, are crucial for adhesiveness of CD11b. In hypercholesterolemic patients, adhesion of isolated monocytes to endothelium ex vivo was dramatically increased over values in healthy control subjects. Treatment of these patients with the HMG-CoA reductase inhibitors lovastatin or simvastatin (20 to 40 mg/day) for 6 weeks slightly decreased total and LDL cholesterol plasma levels and monocyte CD11b surface expression but resulted in a significant reduction of monocyte adhesion to endothelium (p < 0.01, n = 7). CONCLUSIONS: The reduction of CD11b expression and inhibition of CD11b-dependent monocyte adhesion to endothelium may crucially contribute to the clinical benefit of HMG-CoA reductase inhibitors in CHD, independent of cholesterol-lowering effects. PMID- 9350918 TI - HMG-CoA reductase inhibitors: a new class of anti-inflammatory drugs? PMID- 9350919 TI - Effect of L-arginine on human coronary endothelium-dependent and physiologic vasodilation. AB - OBJECTIVES: We hypothesized that L-arginine would improve abnormal coronary vasodilation in response to physiologic stress in patients with atherosclerosis and its risk factors by reversing coronary endothelial dysfunction. BACKGROUND: Studies have demonstrated that physiologic coronary vasodilation correlates with endothelial function and that L-arginine, the substrate for nitric oxide synthesis, improves the response to acetylcholine (Ach). METHODS: Changes in coronary blood flow and epicardial diameter response to Ach, adenosine and cardiac pacing were measured in 32 patients with coronary atherosclerosis or its risk factors and in 7 patients without risk factors and normal coronary angiograms. RESULTS: Intracoronary L-arginine did not alter baseline coronary vascular tone, but the epicardial and microvascular responses to Ach were enhanced (both p < 0.001). The improvement after L-arginine was greater in epicardial segments that initially constricted with Ach; similarly, L-arginine abolished microvascular constriction produced by higher doses of Ach. Thus, there was a negative correlation between the initial epicardial and vascular resistance responses to Ach and the magnitude of improvement with L-arginine (r = -0.55 and r = -0.50, respectively, p < 0.001). D-Arginine did not affect the responses to Ach, and adenosine responses were unchanged with L-arginine. Cardiac pacing induced epicardial constriction was abolished by L-arginine, but microvascular dilation remained unaffected. CONCLUSIONS: Thus, L-arginine improved endothelium dependent coronary epicardial and microvascular function in patients with endothelial dysfunction. Prevention of epicardial constriction during physiologic stress by L-arginine in patients with endothelial dysfunction may be of therapeutic value in the treatment of myocardial ischemia. PMID- 9350920 TI - Normal high energy phosphate ratios in "stunned" human myocardium. AB - OBJECTIVES: We sought to investigate whether alterations in cardiac high energy phosphates occur in postischemic "stunned" human myocardium. BACKGROUND: Transient postischemic myocardial dysfunction is a common phenomenon that occurs in a variety of clinical settings in the absence of necrosis, and its pathogenesis is still unclear. Cardiac high energy phosphates are reduced during ischemia, and persistently altered myocardial high energy phosphate metabolism has been suggested as a mechanism contributing to stunning. METHODS: We studied 29 patients with a first anterior myocardial infarction (MI) who underwent successful reperfusion within 6 h of the onset of chest pain. These patients underwent 31P magnetic resonance spectroscopy (MRS) a mean of 4 days after MI for measurement of left ventricular contractility and relative high energy phosphate metabolites. Twenty-one patients underwent a second 31P MRS study a mean of 39 days after MI. Eight volunteers served as control subjects. RESULTS: Global and infarct area wall motion scores improved significantly between the early and late studies. No difference was found between early cardiac phosphocreatine (PCr)/beta adenosine triphosphate (beta-ATP) ratios in patients and control subjects ([mean +/- SD] 1.51 +/- 0.17 vs. 1.61 +/- 0.18, respectively, p = 0.17) or between early and late study results in patients (1.51 +/- 0.17 vs. 1.53 +/- 0.17, respectively, p = 0.6). For alpha of 0.05, the study had a 90% power to detect a 9% difference. CONCLUSIONS: The results of this study demonstrate normal myocardial PCr/ATP ratios in patients with myocardial stunning after reperfusion and suggest that relative cardiac high energy phosphates are not depleted in stunned human myocardium. PMID- 9350921 TI - Serial changes in response of hibernating myocardium to inotropic stimulation after revascularization: a dobutamine echocardiographic study. AB - OBJECTIVES: We sought to evaluate the serial changes in the response of the hibernating myocardium to dobutamine stimulation after revascularization. BACKGROUND: An improvement in myocardial contraction during dobutamine stress echocardiography (DSE), particularly a biphasic response, predicts recovery of rest function. However, little is known about the changes in the response of the myocardium to dobutamine after revascularization. METHODS: Thirty-four patients with stable coronary artery disease and regional left ventricular dysfunction underwent DSE before, early (within 1 week) and late (>6 weeks) after coronary angioplasty. Dobutamine was given in incremental doses from 2.5 to 40 microg/kg body weight per min. RESULTS: Of 180 revascularized segments with severe rest dysfunction, recovery of rest function was seen in 56 segments (31%) late after angioplasty, 80% of which had early recovery. Ventricular function during DSE was similar early and late after revascularization. Patients who showed a biphasic response to DSE before revascularization (n = 12) had the most improvement in function at rest (mean [+/-SD] wall motion score index [WMSI] 1.98 +/- 0.75 vs. 1.35 +/- 0.54, p < 0.05) and during DSE (2.11 +/- 0.67 vs. 1.21 +/- 0.41, p < 0.05) late after revascularization. Patients with sustained improvement during DSE before revascularization had no significant change in wall motion during DSE after angioplasty. However, patients without improvement in function at low dose DSE, who demonstrated worsening of function at a high dose, had significant augmentation in wall motion during DSE after revascularization (WMSI 2.16 +/- 0.50 vs. 1.60 +/- 0.57, p < 0.05). Patients who had no recovery of rest function had significant improvement in wall motion response to DSE, particularly when ischemia was inducible before revascularization. CONCLUSIONS: In myocardial hibernation, the majority of recovery of rest function occurs early after revascularization. Although patients who recover rest function show the most marked improvement in wall motion during DSE, those without recovery of rest function also have improved function during DSE, particularly when there is evidence of ischemia before revascularization. PMID- 9350922 TI - Assessment of myocardial viability in patients with previous myocardial infarction by using single-photon emission computed tomography with a new metabolic tracer: [123I]-16-iodo-3-methylhexadecanoic acid (MIHA). Comparison with the rest-reinjection thallium-201 technique. AB - OBJECTIVES: We compared the ability of rest single-photon emission computed tomography (SPECT) with [123I]-16-iodo-3-methylhexadecanoic acid (MIHA) and the thallium-201 (Tl-201) rest-reinjection technique to detect myocardial viability after infarction. BACKGROUND: After myocardial infarction, MIHA frequently shows increased uptake in the areas with exercise Tl-201 defects (mismatch), even in patients with an irreversible Tl-201 reinjection defect. Whether such increased uptake is indicative of ischemic but viable myocardium is not known. METHODS: We studied 38 patients who 1) underwent exercise SPECT Tl-201 with rest-reinjection and rest SPECT with MIHA before undergoing percutaneous transluminal coronary angioplasty (PTCA) of an infarct-related coronary artery, and 2) were found to have successful revascularization at follow-up angiography. The relation between SPECT results before PTCA and subsequent improvement in left ventricular wall motion was assessed. RESULTS: A mismatch was evident before PTCA in 51 of 76 infarct-related segments and correlated with subsequent improvement in wall motion (overall accuracy 71%), even for the 27 segments whose exercise defects remained irreversible after Tl-201 reinjection (overall accuracy 81%). The finding of a mismatch clearly enhanced the results provided by the finding of > or = 50% Tl-201 uptake as determined at redistribution (p < 0.05), but not as determined at reinjection, although there was a trend toward a better specificity for the findings of a mismatch. CONCLUSIONS: MIHA is an efficient marker of viability inside exercise-underperfused areas after infarction, even in patients with irreversible Tl-201 reinjection defects. Assessment by conventional SPECT of a mismatch between results obtained with a metabolic tracer (MIHA) and a flow tracer analyzed at exercise (Tl-201) as a marker of myocardial viability is a promising area of research. PMID- 9350923 TI - Blood pressure changes during transient myocardial ischemia: insights into mechanisms. AB - OBJECTIVES: We investigated the contribution of changes in systemic blood pressure to the genesis of spontaneous myocardial ischemia. BACKGROUND: Although increases in heart rate often precede the development of spontaneous myocardial ischemia, it remains a subject of controversy whether these are accompanied by simultaneous changes in blood pressure. METHODS: Using an ambulatory monitoring device that triggered blood pressure recordings from the level of the ST segment, we documented systolic and diastolic blood pressure and heart rate changes related to episodes of ST segment depression in 17 patients with stable coronary artery disease. RESULTS: Systolic blood pressure and heart rate, but not diastolic pressure, increased significantly before the onset of ST segment depression and persisted throughout the ischemic episode. There was a significant correlation between the changes in heart rate and systolic blood pressure during episodes of myocardial ischemia (r = 0.5, p = 0.0005) and between heart rate and systolic blood pressure changes at 1-mm ST segment depression during treadmill exercise testing and ambulatory monitoring (r = 0.73, p = 0.0005 for heart rate; r = 0.77, p = 0.0008 for systolic blood pressure), indicating that patients with a low heart rate threshold during ischemic episodes also had a lower systolic blood pressure threshold before ischemia during both tests. Circadian changes in systolic blood pressure paralleled the variations in heart rate and ischemic episodes, with the lowest values at night. CONCLUSIONS: Significant increases in myocardial oxygen demand, including systolic blood pressure, occur during episodes of spontaneous myocardial ischemia. Patients with a lower heart rate threshold during ischemic episodes had a lower systolic blood pressure threshold during both ambulatory monitoring and treadmill exercise. The effects of antianginal therapy on blood pressure changes during ischemia need to be explored further. PMID- 9350924 TI - Evaluation of exercise thallium scintigraphy versus exercise electrocardiography in predicting survival outcomes and morbid cardiac events in patients with single and double-vessel disease. Findings from the Angioplasty Compared to Medicine (ACME) Study. AB - OBJECTIVES: We sought to evaluate the prognostic ability of cardiac exercise stress tests in predicting cardiac mortality and morbidity in a low risk group of patients with established coronary artery disease (CAD). BACKGROUND: Although previous studies have demonstrated the superior value of stress nuclear cardiac scintigraphy in the prognosis of patients with CAD, none of these studies have focused on patients with a proven angiographic low risk profile (i.e., single- and double-vessel CAD). METHODS: Three hundred twenty-eight patients with documented single- and double-vessel disease were treated by random assignment to percutaneous transluminal coronary angioplasty or medical therapy in the Angioplasty Compared to Medicine (ACME) trial. Six months after randomization, maximal symptom-limited exercise tests were performed with electrocardiography (n = 300) and thallium scintigraphy (n = 270). Patients were followed up for a minimum of 5 years thereafter. RESULTS: A reversible thallium perfusion deficit documented after 6 months of either therapy was associated with an adverse mortality outcome (18% mortality rate with a reversible thallium perfusion defect and 8% mortality rate with no reversible thallium perfusion deficit, p = 0.02). Moreover, an important mortality gradient was demonstrated in relation to the number of reperfusing defects (0 = 7%, 1 to 2 = 15%, >3 = 20%, p = 0.04). Exercise electrocardiography did not predict this mortality outcome. CONCLUSIONS: A reversible thallium perfusion deficit demonstrated 6 months after medical therapy or coronary angioplasty is a valuable prognostic marker in patients with angiographically documented single- and double-vessel disease and is superior to exercise electrocardiography in this regard. PMID- 9350926 TI - Recovery of myocardial perfusion in acute myocardial infarction after successful balloon angioplasty and stent placement in the infarct-related coronary artery. AB - OBJECTIVES: This study sought to investigate changes in myocardial perfusion after direct percutaneous transluminal coronary angioplasty (PTCA) in acute myocardial infarction (MI). BACKGROUND: After initially successful recanalization of the infarct-related artery, coronary perfusion may deteriorate as a result of reocclusion, distal embolization of platelet aggregates formed at the dilated plaque or microvascular reperfusion injury. This change could offset the benefit from early intervention. METHODS: The study included 19 patients in whom the infarct-related artery was successfully recanalized by PTCA with Palmaz-Schatz stent placement within 24 h after the onset of pain. Basal and papaverine-induced coronary blood flow were assessed by Doppler flow velocity measurements and quantitative coronary angiography. In addition, basal and adenosine-induced myocardial blood flow were measured by nitrogen-13 ammonia positron emission tomography (PET). RESULTS: Immediately after completion of the intervention, the average coronary flow reserve (CR) in the recanalized vessel was 1.56 +/- 0.51; it increased to 2.04 +/- 0.65 at 1 h (p = 0.013) and to 2.66 +/- 0.72 at 2 weeks after reperfusion (p = 0.008, n = 16). PET studies in 12 patients revealed that perfusion defect size and CR in the infarct region (2.19 +/- 0.89 vs. 2.33 +/- 0.86) did not change significantly between day 2 after recanalization and 2 weeks. However, we found significant (p < 0.03) increases in basal (by 26%) and adenosine-induced (by 40%) blood flow in the infarct region. CONCLUSIONS: Despite the persistence of a perfusion defect after successful recanalization of the occluded artery in acute MI, CR of the infarct region improves in most patients within 1 h and further improves within 2 weeks. PMID- 9350925 TI - Bivalirudin compared with heparin during coronary angioplasty for thrombus containing lesions. AB - OBJECTIVES: We investigated whether bivalirudin is more effective than heparin in preventing ischemic complications in high risk patients undergoing coronary angioplasty for thrombus-containing lesions detected by angiography. BACKGROUND: Heparin is administered during coronary angioplasty to prevent closure of the dilated vessel. Bivalirudin (Hirulog) is a direct thrombin inhibitor that can be safely substituted for heparin during angioplasty. Bivalirudin has several theoretic advantages over heparin as an anticoagulant agent. METHODS: We performed an observational analysis of the Hirulog Angioplasty Study in which 4,098 patients with unstable or postinfarction angina were randomized to receive either bivalirudin or heparin during coronary angioplasty. The study group for this analysis consisted of 567 patients who had thrombus-containing lesions on angiography. The primary end point was death, myocardial infarction, emergency coronary artery bypass graft surgery or abrupt vessel closure before hospital discharge. RESULTS: Patients with thrombus-containing lesions had a higher incidence of myocardial infarction (5.1% vs. 3.2%, p = 0.03) and abrupt vessel closure (13.6% vs. 8.3%, p < 0.001) than those without thrombus. In patients with thrombus-containing lesions, however, the incidence of the primary end point was not different between the bivalirudin and heparin treatment groups. Furthermore, no difference in the incidence of ischemic events at 6 months was seen between the treatment groups. CONCLUSIONS: Bivalirudin is not more effective than heparin in preventing ischemic complications in patients undergoing coronary angioplasty for thrombus-containing lesions detected by angiography. Other approaches, perhaps involving potent anti-platelet agents, should be considered for patients with thrombus-containing lesions. PMID- 9350927 TI - Long-term follow-up of a high risk cohort after stent implantation in saphenous vein grafts. AB - OBJECTIVES: We sought to provide short- and long-term clinical outcomes of a high risk cohort treated with stents in saphenous vein grafts (SVGs). BACKGROUND: Data on the long-term outcome of SVG stenting in high risk patients are limited. METHODS: Johnson & Johnson stents were implanted in the SVGs of 186 patients (302 stents, 244 lesions). Ninety percent of patients presented with myocardial infarction (MI) or unstable angina (mean +/- SD ejection fraction [EF] 44 +/- 11%, patient age 71 +/- 9 years, graft age 9.4 +/- 5 years). Using a risk score classification, 155 patients (83%) were defined as high risk for repeat surgical repair or angioplasty. RESULTS: The procedural success rate was 97.3%, with 2.7% major complications (death, Q wave MI, coronary artery bypass graft surgery [CABG]). Clinical follow-up was obtained in 177 patients (mean 19.1 +/- 13.5 months, range 7 to 59). Event rates were 10% for death; 9% for MI; 11% for repeat CABG; and 15% for repeat angioplasty (total events 45%). Kaplan-Meier estimated survival and event-free survival at 4 years were 0.79 +/- 0.06 and 0.29 +/- 0.07, respectively. Predictors of death were congestive heart failure (p < 0.01) and EF <44% (p < 0.05). Predictors of combined events of death, MI and CABG were low EF (p < 0.01) and high SVG age (>10 years, p < 0.01). There were 66 revascularization procedures (35% of patients), 24% of which were in nontarget lesions. Fifty-three percent of the cardiac events occurred during the first year of follow-up. Of the 160 survivors, 36% were free of angina, 49% were in Canadian Cardiovascular Society functional class I or II, and 15% were in class III or IV. Sixty-nine percent of patients were in class I or II according to the Specific Activity Scale, and 31% of patients were in class III or IV. CONCLUSIONS: Balloon expandable stent implantation in the SVGs of high risk patients is associated with a low early complication rate. Expected survival rates are good, as are the anginal and functional classifications, but there is a high rate of recurrent events and need for repeat revascularization. Vein graft stenting is an acceptable palliative option in many high risk patients. PMID- 9350928 TI - Percutaneous interventions alter the hemostatic profile of patients with unstable versus stable angina. AB - OBJECTIVES: The objectives of this study were to define the hemostatic profiles of patients with unstable angina compared with patients with stable angina and to investigate the effect of percutaneous interventions on the follow-up hemostatic profiles of these patients. BACKGROUND: Disturbances in hemostatic factors have been shown to be present in various clinical syndromes involving coronary artery disease. However, their role in stable angina versus unstable angina is less well defined. METHODS: We studied 61 patients with either stable or unstable angina undergoing percutaneous coronary interventions. Blood samples were drawn immediately before the intervention and at 1-month follow-up. Plasma levels of tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI 1) and von Willebrand factor (vWF) were measured by enzyme-linked immunosorbent assays. RESULTS: Patients with unstable angina had significantly higher t-PA levels (mean [+/-SE] 23.7 +/- 3.4 vs. 14.3 +/- 1.4 ng/ml, respectively, p = 0.02) and vWF antigen concentrations (2,231 +/- 157 vs. 1,792 +/- 108 mU/ml, respectively, p = 0.03) than patients with stable angina. No statistically significant differences were observed in the PAI-1 levels between the two groups (27.9 +/- 5.5 vs. 21.4 +/- 2.5 ng/ml, respectively, p = 0.25). At 1-month follow up, there were no longer any significant differences in the t-PA or vWF levels between the two groups (15.7 +/- 1.2 vs. 13.6 +/- 0.6 ng/ml, p = 0.13; 1,962 +/- 170 vs. 1,809 +/- 88 mU/ml, p = 0.39, respectively). There were no significant differences between the hemostatic profiles of patients undergoing percutaneous transluminal coronary angioplasty or coronary stenting initially and at 1-month follow-up. CONCLUSIONS: These data suggest that elevated plasma levels of t-PA and vWF may correlate with instability of atheromatous plaques, and that their decrease after coronary interventions may reflect plaque reendothelialization and stabilization. PMID- 9350929 TI - Left ventricular diastolic filling with an implantable ventricular assist device: beat to beat variability with overall improvement. AB - OBJECTIVES: We studied the effects of left ventricular (LV) unloading by an implantable ventricular assist device on LV diastolic filling. BACKGROUND: Although many investigators have reported reliable systemic and peripheral circulatory support with implantable LV assist devices, little is known about their effect on cardiac performance. METHODS: Peak velocities of early diastolic filling, late diastolic filling, late to early filling ratio, deceleration time of early filling, diastolic filling period and atrial filling fraction were measured by intraoperative transesophageal Doppler echocardiography before and after insertion of an LV assist device in eight patients. A numerical model was developed to simulate this situation. RESULTS: Before device insertion, all patients showed either a restrictive or a monophasic transmitral flow pattern. After device insertion, transmitral flow showed rapid beat to beat variation in each patient, from abnormal relaxation to restrictive patterns. However, when the average values obtained from 10 consecutive beats were considered, overall filling was significantly normalized from baseline, with early filling velocity falling from 87 +/- 31 to 64 +/- 26 cm/s (p < 0.01) and late filling velocity rising from 8 +/- 11 to 32 +/- 23 cm/s (p < 0.05), resulting in an increase in the late to early filling ratio from 0.13 +/- 0.18 to 0.59 +/- 0.38 (p < 0.01) and a rise in the atrial filling fraction from 8 +/- 10% to 26 +/- 17% (p < 0.01). The deceleration time (from 112 +/- 40 to 160 +/- 44 ms, p < 0.05) and the filling period corrected by the RR interval (from 39 +/- 8% to 54 +/- 10%, p < 0.005) were also significantly prolonged. In the computer model, asynchronous LV assistance produced significant beat to beat variation in filling indexes, but overall a normalization of deceleration time as well as other variables. CONCLUSIONS: With LV assistance, transmitral flow showed rapidly varying patterns beat by beat in each patient, but overall diastolic filling tended to normalize with an increase of atrial contribution to the filling. Because of the variable nature of the transmitral flow pattern with the assist device, the timing of the device cycle must be considered when inferring diastolic function from transmitral flow pattern. PMID- 9350930 TI - Evaluation of acute dual-chamber pacing with a range of atrioventricular delays on cardiac performance in refractory heart failure. AB - OBJECTIVES: This study evaluated how variations in atrioventricular (AV) delay affect hemodynamic function in patients with refractory heart failure being supported with intravenous inotropic and intravenous or oral inodilating agents. BACKGROUND: Although preliminary data have suggested that dual-chamber pacing with short AV delays may improve cardiac function in patients with heart failure, detailed Doppler and invasive hemodynamic assessment of patients with refractory New York Heart Association class IV heart failure has not been performed. METHODS: Nine patients with functional class IV clinical heart failure had Doppler assessment of transvalvular flow and right heart catheterization performed during pacing at AV delays of 200, 150, 100 and 50 to 75 ms. RESULTS: Systemic arterial, pulmonary artery, right atrial and pulmonary capillary wedge pressures, cardiac index, systemic and pulmonary vascular resistances, stroke volume index, left ventricular stroke work index (SWI) and arteriovenous oxygen content difference demonstrated no significant changes during dual-chamber pacing with AV delays of 200 to 50 to 75 ms. There were also no changes in the Doppler echocardiographic indexes of systolic or diastolic ventricular function. The study was designed with SWI as the outcome variable. Assuming a clinically significant change in the SWI of 5 g/min per m2, a type I error of 0.05 and the observed standard deviation from our study, the observed power of our study is 85% (type II error of 15%). CONCLUSIONS: Changes in AV delay between 200 and 50 ms during dual-chamber pacing do not significantly affect acute central hemodynamic data, including cardiac output and systolic or diastolic ventricular function in patients with severe refractory heart failure due to dilated cardiomyopathy. PMID- 9350931 TI - Stress-induced left ventricular outflow tract obstruction: a potential cause of dyspnea in the elderly. AB - OBJECTIVES: We sought to identify the pattern of disturbed left ventricular physiology associated with symptom development in elderly patients with effort induced breathlessness. BACKGROUND: Limitation of exercise tolerance by dyspnea is common in the elderly and has been ascribed to diastolic dysfunction when left ventricular cavity size and systolic function appear normal. METHODS: Dobutamine stress echocardiography was used in 30 patients (mean [+/-SD] age 70 +/- 12 years; 21 women, 9 men) with exertional dyspnea and negative exercise test results, and the values were compared with those in 15 control subjects. RESULTS: Before stress, left ventricular end-diastolic and end-systolic dimensions were reduced, fractional shortening was increased, and the basal septum was thickened (2.3 +/- 0.5 vs. 1.4 +/- 0.2 cm, p < 0.001, vs. control subjects) in the patients, but posterior wall thickness did not differ from that in control subjects. Left ventricular outflow tract diameter, measured as systolic mitral leaflet septal distance, was significantly reduced (13 +/- 4.5 vs. 18 +/- 2 mm, p < 0.001). Isovolumetric relaxation time was prolonged, and peak left ventricular minor axis lengthening rate was reduced (8.1 +/- 3.5 vs. 10.4 +/- 2.6 cm/s, p < 0.05), suggesting diastolic dysfunction. Transmitral velocities and the E/A ratio did not differ significantly. At peak stress, heart rate increased from 66 +/- 8 to 115 +/- 20 beats/min in the control subjects, but blood pressure did not change. Transmitral A wave velocity increased, but the E/A ratio did not change. Left ventricular outflow tract velocity increased from 0.8 +/- 0.1 to 2.0 +/- 0.2 m/s, and mitral leaflet septal distance decreased from 18 +/- 2 to 14 +/- 3 mm, p < 0.001. In the patients, heart rate rose from 80 +/- 12 to 132 +/- 26 beats/min and systolic blood pressure from 143 +/- 22 to 170 +/- 14 mm Hg (p < 0.001 for each), but left ventricular dimensions did not change. Peak left ventricular outflow tract velocity increased from 1.5 +/- 0.5 m/s (at rest) to 4.2 +/- 1.2 m/s; mitral leaflet septal distance fell from 13 +/- 4.5 to 2.2 +/- 1.9 mm (p < 0.001); and systolic anterior motion of mitral valve appeared in 24 patients (80%) but in none of the control subjects (p < 0.001). Measurements of diastolic function did not change. All patients developed dyspnea at peak stress, but none developed a new wall motion abnormality or mitral regurgitation. CONCLUSIONS: Although our patients fulfilled the criteria for "diastolic heart failure," diastolic dysfunction was not aggravated by pharmacologic stress. Instead, high velocities appeared in the left ventricular outflow tract and were associated with basal septal hypertrophy and systolic anterior motion of the mitral valve. Their appearance correlated closely with the development of symptoms, suggesting a potential causative link. PMID- 9350932 TI - Valve excrescences: prevalence, evolution and risk for cardioembolism. AB - OBJECTIVES: We sought to determine prospectively the prevalence, evolution and embolic risk of valve excrescences in normal subjects and patients with and without suspected cardioembolism. BACKGROUND: Valve excrescences detected by transesophageal echocardiography (TEE) have been considered a cardioembolic substrate in selected patients. METHODS: Ninety healthy volunteers (Group I) and 88 patients without suspected cardioembolism and a normal TEE (Group II) were studied and followed up clinically for 58 +/- 21 and 48 +/- 20 months, respectively. To assess the evolution of valve excrescences, 45 of these subjects underwent repeat TEE at 31 +/- 13 months. The findings in Groups I and II were compared with those of Group III--49 patients referred for TEE for suspected cardioembolism. RESULTS: Valve excrescences were detected in 34 subjects (38%) in Group I and in 41 patients (47%) in Group II. In Group III, 20 patients (41%) had excrescences, but 85% of them had other potential cardiac or vascular sources of embolism. In all groups, mitral valve excrescences were predominant (68% to 76%), followed by aortic (38% to 50%) and right-sided valves (<10%). Excrescences were equally frequent in men and women and between all age groups studied. During follow-up in Groups I and II, excrescences persisted unchanged, and 1 (1.4%) of 74 patients with and 2 (2%) of 99 subjects without excrescences had cerebral ischemic events (80% power to detect a clinically meaningful difference of 4%). CONCLUSIONS: Valve excrescences are common on the left-sided heart valves of normal subjects and patients regardless of gender and age; they persist unchanged over time and do not appear to be a primary source of cardioembolism. PMID- 9350933 TI - Valve excrescences: harmless and common or strokes-in-waiting? PMID- 9350934 TI - Assessing the outcomes of coronary artery bypass graft surgery: how many risk factors are enough? Steering Committee of the Cardiac Care Network of Ontario. AB - OBJECTIVES: We sought to determine whether more comprehensive risk-adjustment models have a significant impact on hospital risk-adjusted mortality rates after coronary artery bypass graft surgery (CABG) in Ontario, Canada. BACKGROUND: The Working Group Panel on the Collaborative CABG Database Project has categorized 44 clinical variables into 7 core, 13 level 1 and 24 level 2 variables, to reflect their relative importance in determining short-term mortality after CABG. METHODS: Using clinical data for all 5,517 patients undergoing isolated CABG in Ontario in 1993, we developed 12 increasingly comprehensive risk-adjustment models using logistic regression analysis of 6 of the Panel's core variables and 6 of the Panel's level 1 variables. We studied how the risk-adjusted mortality rates of the nine cardiac surgery hospitals in Ontario changed as more variables were included in these models. RESULTS: Incorporating six of the core variables in a risk-adjustment model led to a model with an area under the receiver operating characteristic (ROC) curve of 0.77. The ROC curve area slightly improved to 0.79 with the inclusion of six additional level 1 variables (p = 0.063). Hospital risk-adjusted mortality rates and relative rankings stabilized after adjusting for six core variables. Adding an additional six level 1 variables to a risk-adjustment model had minimal impact on overall results. CONCLUSIONS: A small number of core variables appear to be sufficient for fairly comparing risk-adjusted mortality rates after CABG across hospitals in Ontario. For efficient interprovider comparisons, risk-adjustment models for CABG could be simplified so that only essential variables are included in these models. PMID- 9350935 TI - Clinical shock tolerability and effect of different right atrial electrode locations on efficacy of low energy human transvenous atrial defibrillation using an implantable lead system. AB - OBJECTIVES: The objectives of this study were 1) to evaluate the effect of different right atrial electrode locations on the efficacy of low energy transvenous defibrillation with an implantable lead system; and 2) to qualitate and quantify the discomfort from atrial defibrillation shocks delivered by a clinically relevant method. BACKGROUND: Biatrial shocks result in the lowest thresholds for transvenous atrial defibrillation, but the optimal right atrial and coronary sinus electrode locations for defibrillation efficacy in humans have not been defined. METHODS: Twenty-eight patients (17 men, 11 women) with chronic atrial fibrillation (AF) (lasting > or = 1 month) were studied. Transvenous atrial defibrillation was performed by delivering R wave-synchronized biphasic shocks with incremental shock levels (from 180 to 400 V in steps of 40 V). Different electrode location combinations were used and tested randomly: the anterolateral, inferomedial right atrium or high right atrial appendage to the distal coronary sinus. Defibrillation thresholds were defined in duplicate by using the step-up protocol. Pain perception of shock delivery was assessed by using a purpose-designed questionnaire; sedation was given when the shock level was unacceptable (tolerability threshold). RESULTS: Sinus rhythm was restored in 26 of 28 patients by using at least one of the right atrial electrode locations tested. The conversion rate with the anterolateral right atrial location (21 [81%] of 26) was higher than that with the inferomedial right atrial location (8 [50%] of 16, p < 0.05) but similar to that with the high right atrial appendage location (16 [89%] of 18, p > 0.05). The mean defibrillation thresholds for the high right atrial appendage, anterolateral right atrium and inferomedial right atrium were all significantly different with respect to energy (3.9 +/- 1.8 J vs. 4.6 +/- 1.8 J vs. 6.0 +/- 1.7 J, respectively, p < 0.05) and voltage (317 +/- 77 V vs. 348 +/- 70 V vs. 396 +/- 66 V, respectively, p < 0.05). Patients tolerated a mean of 3.4 +/- 2 shocks with a tolerability threshold of 255 +/- 60 V, 2.5 +/- 1.3 J. CONCLUSIONS: Low energy transvenous defibrillation with an implantable defibrillation lead system is an effective treatment for AF. Most patients can tolerate two to three shocks, and, when the starting shock level (180 V) is close to the defibrillation threshold, they can tolerate on average a shock level of 260 V without sedation. Electrodes should be positioned in the distal coronary sinus and in the high right atrial appendage to achieve the lowest defibrillation threshold, although other locations may be suitable for certain patients. PMID- 9350936 TI - Heart rate variability and dispersion of QT interval in patients with vulnerability to ventricular tachycardia and ventricular fibrillation after previous myocardial infarction. AB - OBJECTIVES: This study was designed to compare QT dispersion measured from the standard 12-lead electrocardiogram and 24-h heart rate variability in patients with vulnerability to either ventricular tachycardia or ventricular fibrillation after a previous myocardial infarction. BACKGROUND: Increased QT interval dispersion and reduced heart rate variability have been shown to be associated with vulnerability to ventricular tachyarrhythmias, but the data have mainly been pooled from patients with presentation of stable ventricular tachycardia and ventricular fibrillation. METHODS: QT dispersion and time domain and two dimensional vector analysis of heart rate variability were studied in 30 survivors of ventricular fibrillation with a previous myocardial infarction and with inducible unstable ventricular tachyarrhythmia by programmed electrical stimulation and in 30 postinfarction patients with clinical and inducible stable monomorphic sustained ventricular tachycardia. Both of these patient groups were matched, with respect to age, gender and left ventricular ejection fraction, with an equal number of postinfarction control patients without a history of arrhythmic events or inducible ventricular tachyarrhythmia and arrhythmia-free survival during a follow-up period of 2 years. Forty-five age-matched healthy subjects served as normal control subjects. RESULTS: Standard deviation of all sinus intervals and long-term continuous RR interval variability analyzed from Poincare plots were reduced in patients with vulnerability to ventricular fibrillation (p < 0.001 for both), but not in patients with ventricular tachycardia (p = NS for both), compared with postinfarction control subjects. Corrected QT (QTc) dispersion was significantly broader both in patients with ventricular fibrillation (p < 0.001) and in those with ventricular tachycardia (p < 0.05) than in matched postinfarction control subjects. Heart rate variability performed better than QTc dispersion in predicting vulnerability to ventricular fibrillation. CONCLUSIONS: Increased QT dispersion is associated with vulnerability to both ventricular tachycardia and ventricular fibrillation. Low heart rate variability is specifically related to susceptibility to ventricular fibrillation but not to stable monomorphic ventricular tachycardia, suggesting that the autonomic nervous system modifies the presentation of life-threatening ventricular arrhythmias. PMID- 9350937 TI - Adenosine-sensitive ventricular tachycardia from the anterobasal left ventricle. AB - OBJECTIVES: This study demonstrates that exercise-provocable tachycardia resembling right ventricular outflow tract tachycardia may originate from the anterobasal left ventricle. BACKGROUND: Reentry is the operative mechanism of idiopathic left ventricular tachycardia, with a QRS complex of right bundle branch block and superior axis that is responsive to verapamil but not adenosine. Whether some mechanism other than reentry is operative in some idiopathic left ventricular tachycardias is unclear. METHODS: In 4 of 53 consecutive patients with idiopathic left ventricular tachycardia, the tachycardia was sensitive to adenosine. These four patients were women 63, 61, 61 and 31 years old and were the subjects of the present study. RESULTS: In all four patients, spontaneous tachycardia was related to exercise or emotional stress. The tachycardia displayed atypical left (one patient) or right (three patients) bundle branch block with an inferior axis and marked variation in cycle length. An intravenous bolus of adenosine triphosphate (10 to 20 mg) terminated tachycardia in all four patients. Tachycardia was terminated or prevented in three patients given intravenous or oral verapamil. Atrial or ventricular incremental or extrastimulus testing induced tachycardia in all four patients (three with, one without isoproterenol infusion). Electrically induced tachycardia also demonstrated marked variation in cycle length, which ranged from 230 to 390 ms. Entrainment was not demonstrable with overdrive pacing from multiple sites. Endocardial mapping during tachycardia revealed that the earliest activations were registered 25, 40, 35 and 50 ms before onset of the QRS complex, respectively, from the anterior aspect of the left ventricle just below the mitral annulus, adjacent to the left ventricular outflow tract. High frequency Purkinje spikes were not recorded at this site. Radiofrequency current delivered to this site successfully ablated the tachycardia in three of the four patients. CONCLUSIONS: Exercise provocable, catecholamine-mediated, verapamil-responsive, adenosine-sensitive ventricular tachycardia may arise from the anterobasal left ventricle adjacent to the outflow tract. PMID- 9350938 TI - Definition of predicted effective antiarrhythmic drug therapy for ventricular tachyarrhythmias by the electrophysiologic study approach: randomized comparison of patient response criteria. AB - OBJECTIVES: We sought to compare efficacies of therapy for ventricular tachyarrhythmias selected by programmed stimulation using two different patient response efficacy criteria: <5 versus <16 repetitive ventricular responses. BACKGROUND: Therapy selection for ventricular tachyarrhythmias by programmed stimulation requires definition of a patient response that predicts long-term efficacy. Such definitions have not been previously compared prospectively. METHODS: Patients with sustained ventricular tachyarrhythmias were randomized to therapy selection using either the <5 or <16 repetitive response criterion of predicted effective therapy. The primary end point was sudden death or recurrence of ventricular tachyarrhythmia requiring intervention. RESULTS: Predicted effective drug therapy was found for 23 (34%) of 68 patients randomized to the <5 criterion and 29 (36%) of 81 patients randomized to the <16 criterion (p = NS). Definition of therapy required 3.0 +/- 1.6 drug trials (mean +/- SD) in patients randomized to the <5 criterion and 2.9 +/- 1.8 trials in patients randomized to the <16 criterion (p = NS). Patients randomized to the <5 criterion had a lower 2 year probability of the primary end point (0.20 +/- 0.05) than did patients randomized to the <16 criterion (0.33 +/- 0.05, one-tailed p = 0.004). The advantage of the <5 criterion was also seen in subgroup analyses involving patients with and without an initial drug efficacy prediction. CONCLUSIONS: The programmed stimulation approach to the selection of antiarrhythmic therapy for ventricular tachyarrhythmias using a patient response criterion of <5 repetitive ventricular responses results in a lower probability of recurrence of ventricular tachyarrhythmia than does use of a <16 repetitive response criterion. PMID- 9350939 TI - Cardiac troponin T in patients with clinically suspected myocarditis. AB - OBJECTIVES: The present study investigated whether myocyte injury can be assessed sensitively by measurement of serum levels of cardiac troponin T (cTnT) in patients with clinically suspected myocarditis and whether cTnT levels may predict the results of histologic and immunohistologic analysis of endomyocardial biopsy specimens. BACKGROUND: Conventionally used laboratory variables often fail to show myocyte injury in patients with clinically suspected myocarditis, possibly because of a low extent of myocardial injury in these patients. Sensitive variables for myocyte injury have not yet been investigated. METHODS: Eighty patients with clinically suspected myocarditis were screened for creatine kinase (CK) activity, MB isoform of CK (CK-MB) activity and cTnT. Endomyocardial biopsy specimens were examined histologically and immunohistologically. RESULTS: cTnT was elevated in 28 of 80 patients with clinically suspected myocarditis, CK in 4 and CK-MB in 1. Histologic analysis alone of the endomyocardial biopsy specimen revealed evidence of myocarditis in only five patients, all with elevated cTnT levels. Twenty-three of 28 patients with elevated cTnT levels had histologically negative findings for myocarditis. Additional immunohistologic analysis revealed evidence of myocarditis in 26 (93%) of 28 patients with elevated cTnT levels and in 23 (44%) of 52 patients with normal cTnT levels. Mean cTnT levels were higher in patients with myocarditis proved histologically or immunohistologically, or both, than in patients without myocarditis (0.59 +/- 1.68 vs. 0.04 +/- 0.05, p < 0.001). CONCLUSIONS: Measurement of serum levels of cTnT provides evidence of myocyte injury in patients with clinically suspected myocarditis more sensitively than does conventional determination of cardiac enzyme levels. Myocardial cell damage may be present even in the absence of histologic signs of myocarditis. Additional immunohistologic analysis often shows lymphocytic infiltrates in these patients. Elevated levels of cTnT are highly predictive for myocarditis in this group. PMID- 9350940 TI - Automatic quantitation of regional myocardial wall motion and thickening from gated technetium-99m sestamibi myocardial perfusion single-photon emission computed tomography. AB - OBJECTIVES: We developed an automatic quantitative algorithm for the measurement of regional myocardial wall motion and wall thickening from three-dimensional gated technetium-99m sestamibi myocardial perfusion single-photon emission computed tomographic images. BACKGROUND: The algorithm measures the motion of the three-dimensional endocardial surface using a modification of the centerline method, as well as wall thickening using both geometry (gaussian fit) and partial volume (counts). METHODS: The algorithm was tested using a "variable thickness" heart phantom, and the quantitative results were compared with visual segmental assessment of myocardial motion and thickening in 79 clinical patients with a wide range of ejection fractions (6% to 87%). RESULTS: Phantom measurements of simulated motion and thickening were accurate regardless of the camera used (dual or triple detector), the angular span of reconstructed data (180 degrees or 360 degrees), the amount of motion (3 or 6 mm) or the amount of thickening (33%, 50% or 100%). Quantitative measurements of segmental motion and thickening in the patients were correlated with visual scores (r = 0.668, exact agreement 72.6%, kappa 0.433 and r = 0.550, exact agreement 74.7%, kappa 0.408, respectively). Significant inverse linear relations exist between the global (summed) visual motion score and the average quantitative motion, and between the global (summed) visual thickening score and the average quantitative thickening. Automatic quantitative ejection fraction measurements correlated extremely well with average quantitative motion (r = 0.929) and thickening (r = 0.959). CONCLUSIONS: Our algorithm is accurate and may be the first automatic technique for the quantitative three-dimensional assessment of regional ventricular function in cardiology. PMID- 9350941 TI - Sustained ventricular tachycardia in adult patients late after repair of tetralogy of Fallot. AB - OBJECTIVES: We sought to determine the features associated with sustained monoform ventricular tachycardia (VT) in adult patients late after repair of tetralogy of Fallot (TOF) and to review their management. BACKGROUND: Patients with repair of TOF are at risk for sudden death. Risk factors for ventricular arrhythmia have been identified from patients with ventricular ectopic beats because of the low prevalence of sustained VT. METHODS: From a retrospective chart review of patients assessed between January 1990 and December 1994, 18 adult patients with VT were identified and compared with 192 with repaired TOF free of sustained arrhythmia. RESULTS: There was no significant difference in age at repair, age at follow-up or operative history. Patients with VT had frequent ventricular ectopic beats (6 of 9 vs. 21 of 101), low cardiac index ([mean +/- SD] 2.4 +/- 0.4 vs. 3.0 +/- 0.8) and more structural abnormalities of the right ventricle (outflow tract aneurysms and pulmonary or tricuspid regurgitation) than control patients. Electrophysiologic map-guided operation was performed in 10 of 14 patients who required reoperation. VT has reoccurred in three of these patients. Four patients did not undergo operation (three received amiodarone; one underwent defibrillator implantation). Two patients with VT also had severe heart failure and died. CONCLUSIONS: Most patients with VT late after repair of TOF have outflow tract aneurysms or pulmonary regurgitation, or both. These patients have a greater frequency of ventricular ectopic beats than arrhythmia-free patients after repair of TOF. A combined approach of correcting significant structural abnormalities (pulmonary valve replacement or right ventricular aneurysmectomy, or both) with intraoperative electrophysiologic-guided ablation may reduce the potential risk of deterioration in ventricular function and enable arrhythmia management to be optimized. PMID- 9350942 TI - Long-term survival in patients with repair of tetralogy of Fallot: 36-year follow up of 490 survivors of the first year after surgical repair. AB - OBJECTIVES: We sought to analyze risk factors for long-term survival (up to 36 years) after surgical repair of tetralogy of Fallot (TOF). BACKGROUND: Survival after repair is excellent, but data >20 years are rare. METHODS: From 1958 to 1977, 658 patients underwent correction of TOF at our institution and were analyzed for survival. Of this patient group (age 12.2 +/- 8.6 years [mean +/- SD], range 2 to 67), 39.7% had a previous palliation. Operative (n = 139) and 1 year (n = 29) deaths were excluded for long-term calculations, resulting in a study group of 490 patients. RESULTS: Actuarial 10-, 20-, 30- and 36-year survival rates were 97%, 94%, 89% and 85%, respectively. Mortality increased 25 years postoperatively from 0.24%/year to 0.94%/year (p = 0.003). The most common cause of death was sudden death (n = 13), followed by congestive heart failure (n = 6). Multivariate correlates of impaired long-term survival were date of operation (before 1970, p = 0.0104), preoperative polycythemia (p = 0.0487) and use of a right ventricular (RV) outflow patch (p = 0.0079). Postoperative systolic RV/left ventricular pressure ratio and age showed no influence. Patients without preoperative polycythemia and an RV outflow patch (n = 164) had a 36-year actuarial survival rate of 96% and normal life expectancy. CONCLUSIONS: Cyanosis, operative experience of the surgeon and an RV outflow tract patch influence long term outcome after repair of TOF in older children. Early repair by experienced surgeons to avoid polycythemia and excessive RV hypertrophy is supported by this study. However, mortality risk increases 25 years postoperatively, and thus heart monitoring should be intensified. PMID- 9350943 TI - Ventricular arrhythmia after repair of tetralogy of Fallot. PMID- 9350945 TI - Vena contracta imaged by Doppler color flow mapping predicts the severity of eccentric mitral regurgitation better than color jet area: a chronic animal study. AB - OBJECTIVES: This study sought to evaluate the relation between the color Doppler imaged vena contracta and the severity of mitral regurgitation (MR) in a chronic animal model of MR. BACKGROUND: The vena contracta, which is defined as the smallest connection between the laminar flow acceleration zone and the turbulent regurgitant jet, has been reported to be a clinically useful marker for evaluating the severity of valvular regurgitation. METHODS: Six sheep with chronic MR produced by previous operation severing the chordae tendineae were examined. MR jet flows and vena contracta widths were imaged using a Vingmed 775 scanner with a 5-MHz transducer. Image data were directly transferred in digital format to a microcomputer for off-line measurement. MR was quantified as peak and mean regurgitant flow rates, regurgitant stroke volumes and regurgitant fractions determined using mitral and aortic electromagnetic flow probes and flowmeters balanced against each other. RESULTS: Vena contracta width correlated well with regurgitant severity determined by electromagnetic flowmeters (r = 0.95, SEE = 0.05 cm, p < 0.0001 for peak regurgitant flow rate; r = 0.85, SEE = 0.08 cm, p < 0.0001 for regurgitant stroke volume; r = 0.90, SEE = 0.07 cm, p < 0.0001 for regurgitant fraction). CONCLUSIONS: This study shows that the vena contracta width method is useful for predicting the severity of MR. It is simple and conveniently available with high resolution equipment. The quantitative comparisons in the present study lay the foundation for future clinical and research studies using this vena contracta technique. PMID- 9350944 TI - Comparison of the effects of selective endothelin ETA and ETB receptor antagonists in congestive heart failure. AB - OBJECTIVES: This study was designed 1) to determine the extent to which endogenous endothelin (ET) affects hemodynamic, hormonal and body fluid balance through ETA and ETB receptors in congestive heart failure (CHF); and 2) to assess the therapeutic benefits and adverse effects of ET receptor antagonists for ETA and ETB on cardiorenal and neurohormonal variables. BACKGROUND: ET has two receptors, ETA and ETB, both of which are distributed in various tissues and cells. In vascular beds, ETA receptors mediate vasoconstriction, whereas ETB receptors mediate vasorelaxation. However, ETB receptors also exist in smooth muscle and mediate vasoconstriction. METHODS: We administered either the ETA receptor antagonist FR139317 (FR [n = 8], 1 and 10 mg/kg body weight) or the ETB receptor antagonist RES-701-1 (RES [n = 8], 0.2 and 1.5 mg/kg) to dogs with CHF induced by rapid ventricular pacing. The effects of both antagonists on cardiorenal and hormonal functions were studied. RESULTS: FR decreased cardiac pressures and the plasma atrial natriuretic peptide (ANP) level and increased cardiac output (CO). Urinary flow rate and urinary sodium excretion increased in association with an increase in the glomerular filtration rate and renal plasma flow (RPF). In contrast, RES increased cardiac pressures and decreased CO. It also decreased the plasma aldosterone level and RPF. Neither antagonist affected plasma norepinephrine levels. CONCLUSIONS: Endogenous ETs increase cardiac pressures and the retention of body fluid through ETA receptors in CHF. The vasodilative action through ETB receptors is overall functionally more important than the constrictive action through ETB receptors. ETs may regulate the secretion of ANP and aldosterone. Our findings suggest that selective ETA receptor antagonists have potential therapeutic benefits affecting both hemodynamic variables and diuresis, whereas ETB receptor antagonists have adverse hemodynamic effects, with the possibility of preventing fluid retention through suppression of aldosterone secretion in dogs with CHF. PMID- 9350946 TI - Effect of significant two-vessel versus one-vessel coronary artery stenosis on myocardial contrast defects observed with intermittent harmonic imaging after intravenous contrast injection during dobutamine stress echocardiography. AB - OBJECTIVES: We sought to determine the effect of multivessel as opposed to single vessel coronary artery stenosis on myocardial contrast defects observed with intermittent harmonic imaging and intravenous perfluorocarbon-exposed sonicated dextrose albumin contrast injection. BACKGROUND: Intermittent harmonic imaging has permitted the detection of myocardial perfusion abnormalities with an intravenous ultrasound contrast agent. The effect of multivessel disease on inducibility of these perfusion abnormalities is unknown. METHODS: In 10 dogs, intravenous injections of contrast agent were given at rest and during dobutamine stress echocardiography when a single coronary artery stenosis was present (> or = 50% diameter by quantitative angiography) and again when a second stenosis (range 44% to 92% diameter) was present in the vessel supplying the adjacent perfusion bed. The peak myocardial contrast was visually and quantitatively assessed in the mid and lateral regions of the perfusion bed of the first stenosis (original stenosis zone) in the presence of one- and two-vessel stenosis. RESULTS: Peak myocardial contrast defects in both the mid and lateral segments of the original stenosis zone during dobutamine stress echocardiography was significantly lower when two-vessel stenosis was present (p = 0.015), especially in the lateral segment. The spatial extent of the perfusion defect in the original stenosis zone risk area increased significantly when two-vessel stenosis was present, and correlated closely with actual risk area (r = 0.99). Previous total occlusion followed by reperfusion of the vessel supplying the original stenosis zone significantly increased the amount of collateral activity between perfusion beds. CONCLUSIONS: Collateral flow limits the spatial extent of inducible ischemia within the risk area of single-vessel stenosis. Restoring blood flow to one perfusion bed reduces the extent of a perfusion abnormality that can be induced in an adjacent stenosed bed. PMID- 9350948 TI - President's page: participate in the "cure". PMID- 9350947 TI - Myocardial cell death and apoptosis in hibernating myocardium. AB - OBJECTIVES: This study was designed to study apoptosis in hypoperfused hibernating myocardium subtending severe coronary stenosis. BACKGROUND: Apoptosis contributes to myocyte death in acute myocardial infarction. METHODS: A left anterior descending coronary artery stenosis was created in 13 pigs and maintained for 24 h (n = 4), 7 days (n = 5) and 4 weeks (n = 4) to reduce coronary blood flow by a mean of 34% with severe regional myocardial systolic dysfunction, as documented by echocardiography. Apoptosis was detected with an in situ end-labeling method and confirmed by "deoxyribonucleic acid laddering" on agarose-gel electrophoresis. The severity of apoptosis was expressed as the percentage of apoptotic myocyte nuclei and nonapoptotic myocardial nuclei. RESULTS: Myocardial blood flow of the anterior left ventricular wall was reduced from 1.00 +/- 0.18 to 0.66 +/- 0.21 ml/min per g (p < 0.01), with a severe reduction of anterior regional wall thickening from a mean (+/-SD) of 39 +/- 4% to 9 +/- 8% (p < 0.01). There was no myocardial infarction in five pigs and minimal patchy infarction of < or = 6% of the area at risk in eight pigs. Apoptotic myocytes were observed in the hibernating myocardial region in all pigs (4.8 +/- 2.3%). Myocyte apoptosis was patchy in distribution and was found predominantly in the subendocardial myocardium (9.8 +/- 4.6%) and rarely in the subepicardial myocardium (0.32 +/- 0.45%). Apoptosis was found not only around focal fibrosis areas, but also in areas without fibrosis or patchy infarction. Apoptosis was found not only in 24-h hypoperfused myocardium, but also in 4-week hypoperfused myocardium. The severity of myocyte apoptosis correlated significantly with regional coronary blood flow reduction (r = 0.75, p < 0.01). No apoptosis was found in the normal control region. CONCLUSIONS: This study demonstrates that there is ongoing myocyte death through myocyte apoptosis in hypoperfused hibernating myocardium. PMID- 9350949 TI - Folie a deux in Japan -- analysis of 97 cases in the Japanese literature. AB - In order to clarify the characteristics of folie a deux in Japan, we examined a total of 97 cases of folie a deux in the Japanese literature covering a period of 90 years, and compared them with the cases reported in Western countries. About 75% of the Japanese cases occur in two individuals, and of these are family cases. The most common combinations are mother-child and married couple. Mother child combinations are much more common than father-child combinations. Female subjects are more often involved than males. The most common diagnosis for the dominant partner is schizophrenia, and the most common diagnosis for the submissive partner is paranoid reaction. Delusion is the most common symptom shared by both partners in Japan. Comparing these Japanese cases to Western ones, sister-sister combinations are less frequent, younger subjects influence the older ones more, and acute religious delusion is more common in Japan than in Western countries. PMID- 9350951 TI - Depressive symptoms among international exchange students, and their predictors. AB - The study described in the present paper prospectively examined the depressive symptoms displayed by a cohort of Japanese high school students (n=144) before and during their 1-year placements with volunteer host families in various countries under the aegis of an international cultural exchange programme. The subjects' level of depression showed a statistically significant increase 6 months after such a placement, but had returned to pre-departure levels by the completion of their 1-year placement. The variables that significantly predicted depressive symptoms during the placement in multiple regression analyses were the neuroticism score and the depressive symptoms measured prior to departure. However, the parental rearing practices and the fluency in the English language did not show a significant correlation. When measures of social support during the placement were entered, the perceived adequacy of social support was found to make an additional contribution to the prediction of depressive symptoms. PMID- 9350950 TI - Symptoms, standards of living and subjective quality of life: a comparative study of schizophrenic and depressed out-patients. AB - The subjective quality of life (QOL) (i.e. individual evaluation of one's life experiences) has been studied according to a series of different parameters such as resource availability, and sociodemographic and clinical variables, at times yielding contradictory results. Subjective quality of life and standard of life from a selected sample of 45 chronic out-patients (25 schizophrenics, and 20 patients with major depression) were evaluated by means of structured interviews. Statistical analysis revealed that subjective QOL was largely independent of standard of living (so long as basic needs were satisfied), diagnosis, and clinical course of illness, and only partly dependent on sociodemographic variables. No correlation was found between clinically evaluated symptoms (both psychotic and depressive) and subjective QOL. On the contrary, significant correlations were found between self-ratings of depression, depressive cognitive attitudes and subjective ratings of QOL. PMID- 9350952 TI - Prevalence of long-term mental health care utilization in The Netherlands. AB - The objective of this study is to estimate the proportion of the population in The Netherlands who receive long-term care for chronic psychiatric problems. The care needs of this population are assessed in terms both of diagnosis and of specific impairments and disabilities. Data from three surveys and two psychiatric case registers in five different areas of The Netherlands provide an estimate of about 3.5 long-term users of psychiatric care per 1000 members of the population aged 20 years or over. One-third of them receive a diagnosis of schizophrenia and related psychotic disorders. Patients most frequently suffer from impairments of mood and affect, volition and drives. Nearly all patients are disabled in their occupational role (work), and about half of the population have problems with self-care and household tasks. Long-term care is to a large extent (40%) provided in hospitals and sheltered accommodation, and the role of day services is relatively insignificant. PMID- 9350953 TI - The prediction of suicidal intent in depressed patients. AB - The aim of the present study was to examine the relationships between suicidal ideation or suicidal attempts and severity of depression, presence of personality disorders, and sociodemographic factors in a population of depressed in-patients. A total of 338 adult depressed psychiatric in-patients were examined and classified according to DSM-III criteria as having major depression with or without melancholic or psychotic features, adjustment disorder with depressed mood or dysthymic disorder. Scores on the Hamilton Depression Rating Scale (HDRS), Beck Depression Inventory (BDI) and Zung Self-Rating Depression and Anxiety Scales (SDS and SAS) were measured. We found that suicidal ideation was significantly related to severity of depression (according to the HDRS and all self-rating scales), a lower global assessment of functioning the year before hospitalization, and previous psychiatric hospitalizations. The items with the strongest predictive value for suicidal ideation were hopelessness, depressed mood, feelings of guilt, loss of interest and low self-esteem. These symptoms predicted 43% of the variance in suicidal ideation. None of the above predictors of suicidal ideation was related to suicidal attempts. Depressed patients with a personality disorder attempted significantly more suicidal attempts and showed more suicidal ideation than depressed patients without personality disorder. No significant correlations were found between suicidal ideation or suicide attempts and gender, marital status, employment status or psychosocial stressors during the previous 6 months. PMID- 9350955 TI - A comparison of quetiapine and chlorpromazine in the treatment of schizophrenia. AB - A 6-week, double-blind, randomized, multicentre, parallel-group study was conducted to compare the efficacy of quetiapine ('Seroquel') (n=101) with that of chlorpromazine (n=100) in hospitalized patients with acute exacerbation of subchronic or chronic schizophrenia, or schizophreniform disorder. The tolerabilities of the two treatments were also compared. The mean daily doses of quetiapine and chlorpromazine at the end of the study were 407 mg and 384 mg, respectively. Both treatments were effective in the treatment of positive and negative symptoms, with a trend towards superior efficacy for quetiapine. The quetiapine group had a lower incidence of adverse events than the chlorpromazine group, and a low incidence of treatment-emergent extrapyramidal symptoms. Quetiapine was not associated with a sustained increase in serum prolactin. These clinical data support the preclinical profile of quetiapine as an atypical antipsychotic agent. PMID- 9350956 TI - Relationship between neurological 'soft signs' and syndromes of schizophrenia. AB - Past research on the importance of 'soft' neurological signs in schizophrenia has often not examined the relationship between specific groups of neurological signs and different dimensions of schizophrenia psychopathology. Gender differences in the reported relationships have never been explored. In this paper we describe a study of 100 DSM-III-R (65 male and 35 female) schizophrenic patients who were rated for neurological 'soft signs' with the Neurological Evaluation Scale (NES) (1), and for schizophrenic symptomatology with the Scale for Assessment of Negative Symptoms (SANS) and the Scale for Assessment of Positive Symptoms (SAPS). Following a factor analysis of NES items, differential relationships were examined between the five derived NES factors and three well-established dimensions of schizophrenic symptomatology, namely psychomotor poverty, disorganization and reality distortion. Our results failed to show any relationship between NES dimensions and either the reality distortion or disorganization dimensions. There was a modest but differentially significant relationship between psychomotor poverty and an extrapyramidal factor on the NES. This relationship was shown only by male subjects, and was influenced by duration of illness but not by age or neuroleptic medication. On the other hand, female subjects showed a significant relationship between psychomotor poverty and an NES factor reflecting attention and initiative, and between reality distortion and coordination/sequencing of motor activity. These relationships in female subjects were, relative to relationships for male subjects, more independent of the effect of medication and duration of illness. PMID- 9350954 TI - Fertility and schizophrenia: evidence for increased fertility in the relatives of schizophrenic patients. AB - Fertility has been observed to be reduced in patients with schizophrenia, although the disorder was seen to occur at a steady rate in the general population. The hypothesis of increased fertility in the healthy relatives of the patients, which maintained the genetic contribution to the disorder has been proposed but has not received much support. The present study reports the fertility rate in 100 schizophrenic patients and their relatives (grandparents, parents, uncles, aunts and siblings). The fertility of the different family members was compared, taking into account the completion of age of maximum reproductivity, i.e. up to 50 years of age. The trends in fertility rates over three generations of patients' families were compared with those in the general population of India over a corresponding period from 1950 to 2000 AD. The patients were observed to be hypofertile, but their parents showed a higher fertility than all other relatives, as well as the general population. The siblings of the patients also tended to have higher fertility rates than the general population. This increased fertility in parents and sibs, who are the probable carriers of the abnormal gene, could compensate for the reduction in genetic contribution to morbid risk for schizophrenia due to reduced reproductivity of the patients themselves. PMID- 9350957 TI - Hospital Anxiety and Depression Scale (HAD): some psychometric data for a Swedish sample. AB - The Hospital Anxiety and Depression Scale (HAD) was evaluated in a Swedish population sample. The purpose of the study was to compare the HAD with the Beck Depression Inventory (BDI) and Spielberger's State Trait Anxiety Inventory (STAI). A secondary aim was to examine the factor structure of the HAD. The results indicated that the factor structure was quite strong, consistently showing two factors in the whole sample as well as in different subsamples. The correlations between the total HAD scale and BDI and STAI, respectively, were stronger than those obtained using the different subscales of the HAD (the anxiety and depression subscales). As expected, there was also a stronger correlation between the HAD and the non-physical items of the BDI. It was somewhat surprising that the factor analyses were consistently extracting two factors, 'depression' and 'anxiety', while on the other hand both BDI and STAI tended to correlate more strongly with the total HAD score than with the specific depression and anxiety HAD subscales. Nevertheless, the HAD appeared to be (as was indeed originally intended) a useful clinical indicator of the possibility of depression and clinical anxiety. PMID- 9350958 TI - Prevalence of DSM-III-R and ICD-10 psychiatric disorders in a Spanish population of 18-year-olds. AB - The aim of this study was to estimate the current prevalence of DSM-III-R and ICD 10 psychiatric disorders in Spanish 18-year-old members of the general population. Subjects were assessed using the Schedules for Clinical Assessment in Neuropsychiatry (SCAN). Nearly 30% of the study subjects reported at least one current disorder according to ICD-10 criteria, and almost 21% reported at least one current disorder according to DSM-III-R criteria. Women had a significantly higher probability of suffering from any psychiatric disorder than men. The most common disorders were insomnia, dysthymia, major depression and simple phobia. Nearly 40% of the diagnosed subjects had one or more comorbid disorders. Comorbidity was found to be higher among female subjects. Consistent with previous risk factor research, it was found that women had higher rates of mood, anxiety and sleep disorders than men. Good communication between parents and their offspring was found to be a protecting factor for all disorders. PMID- 9350959 TI - Platelet monoamine oxidase activity in relation to alleles of dopamine D4 receptor and tyrosine hydroxylase genes. AB - Human personality characteristics and vulnerability to psychopathology are to a large extent dependent upon genetic factors which have yet to be fully defined. The allele distribution of the dopamine D4 receptor (D4DR) and thrombocyte monoamine oxidase (trbc MAO) activity have both been associated with personality traits which are supposedly related, namely 'sensation seeking' according to Zuckerman and 'novelty seeking' according to Cloninger, respectively. In this report, the D4DR allele distribution and trbc MAO activity were studied in 31 psychiatric patients and 21 control subjects. Trbc MAO activity is a biochemical marker of personality that has been shown to be under strong genetic influence. However, no association between the D4DR alleles and trbc MAO could be observed in this material. To our knowledge, this is the first report comparing these two markers, and based upon the results obtained, we speculate that they may be connected with different types of overlapping personality characteristics. The allele distribution of the tyrosine hydroxylase (TH) gene was also determined. TH is the rate-limiting enzyme in the biosynthesis of catecholamines, and it is believed to be involved in different kinds of psychopathology. No covariation between TH gene alleles and trbc MAO activity or D4DR alleles was observed in this material. PMID- 9350960 TI - Admission rates for schizophrenia in The Netherlands: an urban/rural comparison. AB - This article examines admission rates for schizophrenia in terms of degree of urbanization. Patient data was obtained from the Dutch psychiatric register. The scale of urbanization used had five categories and was utilized to establish the degree of urbanization in each municipality (n=647). Admission rates showed a significant positive correlation with the degree of urbanization in the 15-34 years and 35-54 years age groups for both sexes. The average duration of hospitalization and the average number of readmissions showed no association with the degree of urbanization. Taken as a whole, these utilization rates provide evidence of a much higher incidence of schizophrenia in the most urbanized municipalities of The Netherlands. PMID- 9350961 TI - Dysfunctional parenting and a lifetime history of depression in a volunteer sample of Japanese workers. AB - This study explored whether dysfunctional parenting, as measured by the Parental Bonding Instrument, is linked to a lifetime diagnosis of major depressive disorder (MDD) provided by the Inventory to Diagnose Depression, Lifetime Version, in a non-clinical sample from a non-Western culture. Of a total of 239 Japanese volunteer workers, 22 subjects were diagnosed as having had lifetime MDD. Subjects with lifetime MDD reported a significantly lower 'maternal care' score than did those without lifetime MDD. Logistic regression analysis revealed that subjects who reported a lower 'maternal care' score were at a significantly higher risk for lifetime MDD. The results suggest that the link between dysfunctional parenting and MDD may be independent of cultural differences, lending some support to the hypothesis that dysfunctional parenting in childhood may cause adult depression. PMID- 9350962 TI - Immunocytochemical method for investigating in vivo neuronal oxygen radical induced lipid peroxidation. AB - The investigation of oxygen radical-induced lipid peroxidative neuronal damage in the context of acute and chronic neurodegenerative disorders has been largely limited to the use of ex vivo analytical methodologies. These are often fraught with sensitivity or specificity problems, or they are indirect. Furthermore, none of the analytical methods allow precise anatomical identification of the cells that are undergoing peroxidative injury. This paper describes an immunocytochemical method for localization of central nervous system (CNS) lipid peroxidation (LP) that employs a rabbit-derived antibody raised against malondialdehyde (MDA)-modified rabbit serum albumin (RSA). MDA is a breakdown product of peroxidized membrane polyunsaturated fatty acids that avidly binds to cellular proteins. Using the anti-MDA-RSA, we herein illustrate increased MDA derived immunostaining: (1) in the spinal cord of transgenic familial amyotrophic lateral sclerosis (ALS) mice; and (2) in the selectively vulnerable gerbil hippocampal CA1 region after a 5 min episode of forebrain ischemia and its relationship to the time course of neuronal degeneration. PMID- 9350963 TI - A floating microwire technique for multichannel chronic neural recording and stimulation in the awake freely moving rat. AB - We describe a cheap, and relatively simple, method for the construction and implantation of bundles of six fully-floating 25 microm microwire electrodes in the rat central nervous system (CNS). Wires are stiffened for implantation by temporarily attaching them to a micropipette guide with sucrose which subsequently dissolves in the brain. The associated headpiece connector mates with a plug-in FET which gives high quality recordings, free of movement artefacts, even when used with 3 m of unscreened cable. Different electrode configurations are easily selected and sufficient space is available on the headpiece to accommodate injection guide cannulae. The electrode performance was evaluated in 42 implanted rats where the system was used successfully for long term recording of superior colliculus (SC) deep layer neurons and behavioural responses evoked by electrical stimulation of the same wires. We obtained neural recordings on 81% of the 252 implanted wires, with 79% of these providing good, reliably discriminable, single unit responses following post-operative recovery. During a five-week testing period on a subsample of the 42 'best' electrodes (one per animal), we found the same sensory or motor response 1 week after initial testing in 91% wires, 67% after 2 weeks and 24% after 5 weeks. Using waveform templating techniques we were able to show that 62% of the electrodes still working at 5 weeks were reliably recording the same cell. PMID- 9350964 TI - Transvenous versus perinervous stimulation of the phrenic nerve to assess the diaphragmatic strength in rabbits. AB - Diaphragmatic strength can be measured by transdiaphragmatic pressure during phrenic nerve stimulation. In order to avoid phrenic nerve dissection, a transjugular approach of the phrenic nerve can be performed. The objective of this study was to verify the identity of perinervous and transvenous techniques of phrenic nerve stimulation to assess diaphragmatic force. In intact (n = 9) or right phrenicotomized (n = 12) rabbits, we compared esophageal pressure (Peso) induced by supramaximal perinervous stimulation of the phrenic nerve with that obtained by transvenous stimulation of the phrenic nerve. Electromyography (EMG) of the thoracic muscles was studied in four animals. We found no difference between Peso induced by perinervous (PNS) and transvenous (TVS) unilateral or bilateral phrenic nerve stimulation. During unilateral stimulation, no EMG activity was recorded in the non stimulated diaphragm, or in the middle part of the esophagus, or in ipsi- and contralateral accessory inspiratory muscles. We conclude that in rabbits, unilateral or bilateral TVS of the phrenic nerve is functionally equivalent to PNS, whatever the side of stimulation; Peso is not altered by esophageal contraction in TVS. Transvenous stimulation can replace perinervous stimulation in experimental studies, when cervical access is difficult. PMID- 9350965 TI - A method to biotinylate and histochemically visualize ibotenic acid for pharmacological inactivation studies. AB - Ibotenic acid (IA) and kainic acid (KA) are commonly used tools to selectively inactivate neuronal perikarya, eventually leading to their degeneration, without affecting fibers of passage. Reversible inactivations and experimental paradigms that do not allow for long survival times, however, do not permit for histological verification of the site and extent of the lesion by identifying the area showing gliosis. We describe here a method in which IA and KA were conjugated with biotin and thus could be easily visualized histochemically. We pressure-injected biotinylated IA and KA into various hindbrain areas of the electrosensory system in electric fish while monitoring neuronal responses at the injection site and assessing effects on the behavior. Whereas the effects of biotinylated IA did not differ from those of the unbiotinylated form, biotinylated KA lost its physiological activity. Thus, only biotinylated IA could be used successfully. The size of the gliosis seen after a survival time of seven days was similar to the extent of biotin label observed after injection of comparable volumes of biotinylated IA. Moreover, this method resulted in labeling of individual neurons presumably affected by IA and yielded information about their projection patterns which was comparable to labeling seen after intracellular injections of neurobiotin or biocytin. PMID- 9350966 TI - Storing analog data in a video record. AB - We have designed a simple device that will encode, in machine readable form, multiple analog data in a video record. The analog data is visible to the user within the video frame permitting visual correlation of these signals with activity observed in the image. By superimposition of both analog and video data sets, the two are tightly synchronized and remain so in all copies of the data. The analog data can be separated from the image following digitization by a frame grabber. The bandwidth for each of the five analog channels is approximately 5 kHz. The device, which is essentially an eight channel video multiplexor, includes a video channel, a field counter, an amplitude calibration signal and five analog data channels. The amplitude calibrator allows corrections for gain errors that are particularly prevalent when data is stored on video tape. PMID- 9350967 TI - On the accuracy of an [18F]FDOPA compartmental model: evidence for vesicular storage of [18F]fluorodopamine in vivo. AB - The biological accuracy of a nonlinear compartmental model describing the in vivo kinetics of L-3,4-dihydroxy-6-[18F]fluorophenylalanine ([18F]FDOPA) metabolism was investigated. Tissue activities for [18F]FDOPA and its labeled metabolites 3 O-methyl-[18F]FDOPA ([18F]OMFD), 6-[18F]fluorodopamine ([18F]FDA), L-3,4 dihydroxy-6-[18F]fluorophenylacetic acid ([18F]FDOPAC), and 6 [18F]fluorohomovanillic acid ([18F]FHVA) were calculated using a plasma [18F]FDOPA input function, and kinetic constants estimated previously by chromatographic fractionation of 18F-labeled compounds in plasma and brain extracts from rat. Present data accurately reflected the measured radiochemical composition in rat brain for tracer circulation times past 10 min. We formulated the hypothesis that the discrepancy between calculated and measured fractions of [18F]FDOPA and the deaminated metabolite [18F]FDOPAC at times earlier than 10 min reflected storage of [18F]FDA in vesicles without monoamine oxidase. This hypothesis explained the initially rapid appearance of [18F]FDOPAC in striatum by delayed transfer of [18F]FDA from cytosol into vesicles. We conclude that the simpler model of [18F]FDOPA compartmentation is accurate when the cytosolic and vesicular fractions of [18F]FDA are at steady-state; the approach to equilibrium has a time constant of 15-30 min. The present model is valid for positron emission tomography studies of [18F]FDOPA metabolism in living brain. PMID- 9350968 TI - Biotinylated dextran amine and biocytin hydrochloride are useful tracers for the study of retinal projections in the frog. AB - Anatomical study of the topographic organization of retinal projections requires a tracer capable of resolving fine morphological detail and permitting analysis of the projection from either the whole retina or selected areas. To obtain a permanent record of the experiments and to have access to ultrastructural data, it is preferable for the tracer to be compatible with both brightfield microscopy and electron microscopy. Biotinylated dextran amine and biocytin hydrochloride, as employed in the present experiments, meet these needs exceptionally well for anterograde tracing studies on the frog visual system. Both tracers labeled axons and terminal arbors more prominently than comparable material studied by the widely used methods of anterograde fiber-filling with horseradish peroxidase or cobalt. When used to trace the projections from small sectors of retina, the finest unmyelinated fibers in layers A, C and E of the frog optic tectum and their synaptic boutons were made readily visible by the new tracers. PMID- 9350969 TI - An arterially-perfused trunk-hindquarters preparation of adult mouse in vitro. AB - We describe a preparation of arterially-perfused spinal cord with attached hindquarters, taken from adult mouse. This is the first preparation of adult mammalian spinal cord tissue to have the advantages of an in vitro approach whilst retaining intact intraspinal circuitry, sensory inputs, and somatic and sympathetic segmental outputs. The functional integrity of the preparation has been demonstrated by the motor and sympathetic reflexes that can readily be evoked by peripheral noxious thermal, mechanical and electrical stimuli, and also by bladder distension. The mechanical stability of the preparation allows intracellular recordings to be made from spinal dorsal or ventral horn neurones. The intact connectivity permits synaptic responses to be evoked by stimulation of functionally-defined peripheral sensory receptors. The preparation is relatively quick to set up and remains viable for more than 6 h. This model offers the opportunity to perform complex electrophysiological and pharmacological studies on functionally characterised synaptic responses of mature spinal neurones. The choice of the mouse will furthermore permit studies to be performed on genetically mutant strains. PMID- 9350970 TI - Characterization of variables defining hindpaw withdrawal latency evoked by radiant thermal stimuli. AB - We have examined the stability and sources of variation within the nociceptive model of rat hind paw withdrawal from an under-glass radiant stimulus (Hargreaves et al., 1988) using a system where stimulus intensity and floor temperature can be controlled and reproducibly changed. The current study demonstrates that: (i) increased stimulus intensity with a fixed surface temperature is associated with a monotonic decrease in mean response latency and its variance; (ii) for a fixed stimulus intensity, the mean paw withdrawal latency and variance increased as the glass floor temperature is lowered from 30 degrees C to room temperature (25 degrees C). Using subcutaneously-implanted thermocouples and a 30 degrees C glass surface, the subcutaneous paw temperature observed at an interval corresponding to the time at which the animal displayed a paw withdrawal did not differ across multiple heating rates (41-42.5 degrees C). This finding is in agreement with human studies of pain thresholds and C-fiber activity. These studies emphasize the importance of maintaining a fixed surface temperature to reduce experimental variability and the utility of this apparatus across multiple stimulus intensities to define agonist efficacy. PMID- 9350971 TI - On-column monitoring of secretion of catecholamines from single bovine adrenal chromaffin cells by capillary electrophoresis. AB - The secretion of catecholamines from individual bovine adrenal medullary cells was quantitatively monitored by capillary electrophoresis with laser-induced native fluorescence detection. By using a physiological balanced-salt solution as the running buffer for CE, the amount of norepinephrine (NE) and epinephrine (E) secreted by their physiological secretagogue, acetylcholine, and the amount remaining in a single cell can be simultaneously quantified. Among the six different glands (from separate cows) studied, a predominance of E-rich cells were found. There was no apparent relationship between the ratio of NE/E released and the original NE/E content in the cell. The secretion process was also monitored dynamically with this method by continuously passing acetylcholine over the cell during stimulation. From the peak width and shape of the released material, one can estimate the time scale of the release process. PMID- 9350972 TI - A comparison of methods used to detect changes in neuronal discharge patterns. AB - The discharge pattern of two thalamic neurones was recorded from a conscious monkey performing voluntary movements about the wrist joint. The neuronal discharge was displayed as a raster centred on movement of the wrist. The discharge patterns of both neurones was very strongly correlated with movement. Three experienced researchers were asked to examine the data and to classify every part of each trial as background discharge, 'on' (increased firing rate) or 'off' (decreased or zero firing rate) and to mark the times that neuronal discharge changed state. A 'standard output' was made from these classifications. A back-propagation artificial Neural Network (the Network) was used to model the standard output and cumulative sums (CUSUMs) and maximum likelihood was then performed on the data and compared with the Network. There was a high correlation between the output of each observer (r > 0.61) and the standard output and between the Network and the standard output (r > 0.99). However the correlation between standard output and CUSUMs (r = 0.06) and standard output and maximum likelihood (r = 0.36) was much lower. The Network could be trained with as few as 12 trials, indicating a high degree of constancy in the methods employed by the observers. The Network was also highly efficient at detecting changes in state of neuronal activity (r > 0.99). In summary, when used on single trial data, visual inspection is a reliable method for detecting timing of change neuronal discharge and is superior to CUSUM and maximum likelihood. As well it is capable of detecting neuronal discharge state: that is whether firing rate is increased, normal or decreased. Neural Networks promise to be a useful method of confirming the consistency of visual inspection as a means of detecting changes in neuronal discharge pattern. PMID- 9350973 TI - Genomic structure and embryonic expression of the rat type 1 vasoactive intestinal polypeptide receptor gene. AB - The genomic structure of the rat vasoactive intestinal polypeptide type 1 receptor (VIPR I) gene was characterized using polymerase chain reactions (PCR) and DNA sequencing. These studies show that the rat VIPR I gene spans more than 20 kb of DNA sequence and includes thirteen exons separated by twelve introns, ranging from 0.2 to 4.5 kb. VIP is known to play an important role early in embryonic development. To elucidate the receptor-mediated biological functions of VIP in development, the temporal and spatial expression of VIPR I during rat embryonic development was examined in this study. Using in situ hybridization histochemistry with 35S-labeled riboprobe, VIPR I mRNA was detected as early as embryonic day 11 (E11), in the neuroepithelium, foregut, heart and in cells lining the blood vessels. From E14 through E18, VIPR I mRNA was detected in multiple tissues derived from all three germ layers. These include neuronal cells in brain and spinal cord (ectoderm); smooth muscle cells of intestines, cells lining the wall of the heart and blood vessels, kidney, adrenal gland (mesoderm); epithelial cells of airway and of liver (endoderm). The early onset and wide tissue expression of VIPR I suggests that binding of VIP to this receptor may play an important role in rat embryonic development. PMID- 9350974 TI - Evidence that vasoactive intestinal polypeptide is a parasympathetic neurotransmitter in the endocrine pancreas in dogs. AB - Vasoactive intestinal polypeptide (VIP) has been found in pancreatic nerves in several species. Studies were conducted to determine if VIP could be a parasympathetic neurotransmitter in the canine endocrine pancreas. To verify that VIP is localized in pancreatic parasympathetic nerves, sections of canine pancreas were immunostained for VIP. VIP staining was identified in the majority of neuronal cell bodies in intrapancreatic parasympathetic ganglia. In addition. VIP was localized in nerve fibers innervating pancreatic islets in the proximity of alpha cells. Next, to determine if VIP is released during electrical stimulation of parasympathetic nerves, pancreatic spillover of VIP was measured during vagal nerve stimulation (VNS) in anesthetized dogs. VIP spillover increased from a baseline of 630+/-540 pg/min to 2580+/-540 pg/min (delta = +1950+/-490 pg/min, p <0.01). Pancreatic VIP release during VNS was not affected by atropine, whereas ganglionic blockade with hexamethonium nearly abolished the VIP response to VNS (p<0.005 vs control), suggesting that VIP is a postganglionic neurotransmitter in the dog pancreas. To examine the effects of VIP on pancreatic hormone secretion, synthetic VIP was infused locally into the pancreatic artery. VIP, at a low dose (5 pmol/min), increased glucagon secretion from 1750+/-599 to 3800+/-990 pg/min (delta = +2060+/-870 pg/min, p<0.05), but did not affect insulin secretion (delta = -1030+/-760 microU/min, NS). Thus, VIP is contained in and released from pancreatic parasympathetic nerves in proximity to islet alpha cells and exogenous VIP, at a dose which approximates the increase of VIP spillover during VNS, preferentially stimulates glucagon vs insulin secretion. Therefore, VIP is likely to function as a parasympathetic neurotransmitter in the endocrine pancreas in dogs. We hypothesize that VIP could mediate the glucagon response to parasympathetic activation which has been shown to resistant to cholinergic blockade with atropine in several species. PMID- 9350975 TI - Comparison of Fos induced in rat brain by GLP-1 and amylin. AB - The patterns of Fos-like immunoreactivity (Fos-ir) in rat brain were compared following treatment of rats with two anorectic 'gut' peptides. Central administration of GLP-1 produced dose-related increases in Fos-ir in the area postrema (AP) and caudal nucleus of the solitary tract (cNTS) as well as strong activation in the lateral parabrachial nucleus (LPBE), hypothalamic paraventricular nucleus (PVN), bed nucleus of the stria terminalis (BNST) and central nucleus of the amygdala (CeA). At centrally-active doses, peripheral administration of GLP-1 did not induce Fos-ir in brain. In contrast, peripheral administration of amylin produced strong Fos-ir in the AP and cNTS, as well as the BNST and CeA, but not in the PVN. The common activation of the LPB-BNST-CeA by these and other previously-studied anorectics suggest this is an important circuit involved in satiety. PMID- 9350976 TI - The AT4 receptor agonist [Nle1]-angiotensin IV reduces mechanically induced immediate-early gene expression in the isolated rabbit heart. AB - Angiotensin II (ANG II), acting principally at the AT1 receptor, modulates mechanically-induced cardiac growth. The ANG II metabolite Angiotensin IV (ANG IV) has been shown to inhibit ANG II-induced mRNA and protein synthesis in chick cardiomyocytes. This effect did not involve the AT1 receptor, but was likely an action at the AT4 receptor. To determine if ANG IV also modulates a mechanically induced cardiac growth response, we studied the effects of two AT4 receptor ligands, [Nle1]-ANG IV and [divalinal]-ANG IV, on mechanically-induced immediate early gene expression (c-fos, egr-1, and c-jun) in the buffer perfused (30 degrees C), ejecting, isolated rabbit heart. Mechanical load alone (high systolic pressure and high end-diastolic volume) induced approximately 23-, 49- and 5-fold increases in c-fos, egr-1 and c-jun mRNA (in comparison to control hearts). Perfusion with [Nle1]-ANG IV (10[-10] mol/l) reduced the mechanically-induced expression of c-/fos and egr-1 by 42% and 48%, respectively (P < 0.05). Mechanically-induced c-jun expression was not significantly reduced. Perfusion with [divalinal]-ANG IV (10[-8] mol/l) had no effect on mechanically-induced immediate-early gene expression. We conclude that AT4 receptor agonism influences mechanical immediate-early gene expression, and propose the hypothesis that AT1 and AT4 receptors initiate opposing effects on mechanically-induced immediate early gene expression in the isolated rabbit left ventricle. PMID- 9350977 TI - PACAP and VIP inhibit pyloric muscle through VIP/PACAP-preferring receptors. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide with structural homology to vasoactive intestinal polypeptide (VIP). Two receptor types for PACAP have been described: PACAP preferring receptors are selective for PACAP; whereas VIP/PACAP preferring receptors have similar affinity for both PACAP and VIP. Both VIP and PACAP are present in enteric nerves at the pylorus. VIP is known to exert inhibitory effects on pyloric muscle; the effect of PACAP is unknown. The aims of this study were to determine the effect of PACAP on pyloric muscle and to characterize the PACAP receptor. METHODS: Rabbit pyloric muscle strips were cut parallel to circular muscle fibres and placed in muscle baths. The effect of PACAP and VIP were quantitated as percent of basal motility index (MI). RESULTS: PACAP-27, PACAP-38, and VIP had dose dependent inhibitory effects on the spontaneous phasic contractions of the pylorus. The PACAP-27- induced relaxation was inhibited by the PACAP receptor antagonist PACAP6-27, but was not affected by tetrodotoxin. VIP also had dose dependent inhibitory effects on pyloric muscle. The VIP relaxation was inhibited by PACAP6-27, but not affected by tetrodotoxin. CONCLUSIONS: These studies indicate that, similar to VIP, PACAP inhibits pyloric muscle. The inhibitory effect of the PACAP receptor antagonist on both PACAP and VIP-induced relaxation suggest that PACAP and VIP act at the same receptor, a VIP/PACAP preferring receptor. PMID- 9350978 TI - Purification and characterization of islet hormones (insulin, glucagon, pancreatic, polypeptide and somatostatin) from the Burmese python, Python molurus. AB - Insulin was purified from an extract of the pancreas of the Burmese python, Python molurus (Squamata:Serpentes) and its primary structure established as: A Chain: Gly-Ile-Val-Glu-Gln-Cys-Cys-Glu-Asn-Thr10-Cys-Ser-Leu-Tyr-Glu-Leu- Glu-Asn Tyr-Cys20-Asn. B-Chain: Ala-Pro-Asn-Gln-His-Leu-Cys-Gly-Ser-His10-Leu-Val-Glu-Ala Leu-Tyr- Leu-Val-Cys-Gly20-Asp-Arg-Gly-Phe-Tyr-Tyr-Ser-Pro-Arg-Ser30. With the exception of the conservative substitution Phe --> Tyr at position B25, those residues in human insulin that comprise the receptor-binding and those residues involved in dimer and hexamer formation are fully conserved in python insulin. Python insulin was slightly more potent (1.8-fold) than human insulin in inhibiting the binding of [125I-Tyr-A14] insulin to the soluble full-length recombinant human insulin receptor but was slightly less potent (1.5-fold) than human insulin for inhibiting binding to the secreted extracellular domain of the receptor. The primary structure of python glucagon contains only one amino acid substitution (Ser28 --> Asn) compared with turtle/duck glucagon and python somatostatin is identical to that of mammalian somatostatin-14. In contrast, python pancreatic polypeptide (Arg-Ile-Ala-Pro-Val-Phe-Pro-Gly-Lys-Asp10-Glu-Leu Ala-Lys-Phe- Tyr20-Thr-Glu-Leu-Gln-Gln-Tyr-Leu-Asn-Ser-Ile30-Asn-Arg-Pro-Arg Phe.NH2) contains only 35 instead of the customary 36 residues and the amino acid sequence of this peptide has been poorly conserved between reptiles and birds (18 substitutions compared with alligator and 20 substitutions compared with chicken). PMID- 9350979 TI - Isolation and structural elucidation of eight kinins from the retrocerebral complex of the American cockroach, Periplaneta americana. AB - By monitoring the contractile activity of the hindgut of the American cockroach in vitro eight myotropic neuropeptides were isolated from the retrocerebral complex of the American cockroach. Peptide sequence analysis and mass spectrometry yielded the following structures: Arg- Pro-Ser-Phe-Asn-Ser-Trp-Gly NH2 (Pea-K-1), Asp-Ala-Ser-Phe-Ser-Ser-Trp-Gly-NH2 (Pea-K-2), Asp-Pro-Ser-Phe-Asn Ser-Trp-Gly-NH2 (Pea-K-3), Gly-Ala-Gln-Phe-Ser-Ser-Trp-Gly-NH2 (Pea-K-4), Ser-Pro Ala-Phe-Asn-Ser-Trp-Gly-NH2 (Pea-K-5), Asp-Pro-Ala-Phe-Ser-Ser-Trp-Gly-NH2 (Lem-K 7), Gly-Ala-Asp-Phe-Tyr-Ser-Trp-Gly-NH2 (Lem-K-8) and Ala-Phe-Ser-Ser-Trp-Gly-NH2 (Lom-K). The C-terminal sequence Phe-X-Ser-Trp-Gly-NH2 characterized the peptides as members of the insect kinin family. All structures were confirmed by comparison of retention times between synthetic and natural peptides. The threshold concentration for stimulatory effects of the synthetic peptides on the isolated hindgut was about 10(-9) M and there was no significant difference measured between the different kinin forms. These neuropeptides are the first members of the insect kinin-family isolated from the American cockroach. Their occurrence in the retrocerebral complex suggests a physiological role as neurohormone. PMID- 9350980 TI - The C-terminal domain of the human EP4 receptor confers agonist-induced receptor desensitization in a receptor hybrid with the rat EP3beta receptor. AB - Prostaglandin E2 receptors (EPR), which belong to the family of heterotrimeric G protein-coupled ectoreceptors with seven transmembrane domains, can be classified into four subtypes according to their ligand binding and G protein coupling specificity. Of these, EP3betaR is coupled to Gi, whereas EP4R is coupled to Gs. EP4R, in contrast to EP3betaR, shows agonist-induced desensitization. The C terminal domain and the third intracellular loop of these receptors have been implicated in G protein coupling specificity and desensitization. Here, receptor hybrids consisting of the main portion of rat EP3betaR and either the C-terminal domain or the third intracellular loop of human EP4R were used to study the contribution of the respective receptor domains to G protein coupling and desensitization. Neither the EP4R C-terminal domain nor the EP4R third intracellular loop alone was sufficient to change the coupling specificity of the rEP3hEP4 receptor hybrids from Gi to Gs or to confer additional Gs coupling. However, the EP4R C-terminal domain but not the third intracellular loop was necessary and sufficient to mediate rapid agonist-induced, second messenger independent desensitization in the Gi-coupled hybrid receptors. PMID- 9350981 TI - The bacterial pigment xanthomonadin offers protection against photodamage. AB - Xanthomonas oryzae pv. oryzae is a bacterial pathogen that causes leaf blight, a serious disease of rice. Most members of the genus Xanthomonas produce yellow, membrane bound, brominated aryl polyene pigments called xanthomonadins whose functional role is unclear. We find that pigment-deficient mutants of X. oryzae pv. oryzae exhibit hypersensitivity to photobiological damage. A clone containing the xanthomonadin biosynthetic gene cluster alleviates the hypersensitivity of the pigment-deficient mutant. Extracts containing xanthomonadin provide protection against photodynamic lipid peroxidation in liposomes. These results lead us to suggest a role for the pigment, namely protection against photodamage. PMID- 9350982 TI - Structural requirement of highly-conserved residues in globins. AB - Globins have remarkable sequence diversity, and yet maintain a common fold. In spite of the diversity, there are highly-conserved residues at several sites. The conserved residues were examined in terms of the structural stability, by employing the pseudo-energy functions of the structure/sequence compatibility method. The fitness of each residue type to the structural environment was evaluated at seven highly-conserved sites: the Leu (at the B10 site), Phe (CD1), and Leu (F4) residues were found to fit their respective sites due to hydrophobic interactions; Pro (C2) stabilizes the N-terminal edge of an alpha-helical structure; and Phe (CD4) is stabilized by backbone hydrogen-bonding to Phe (CD1). On the other hand, the other two residues, His (E7) and His (F8), are poorly suited to the sites from a structural viewpoint, suggesting that their conservation clearly results from a heme-related functional requirement. The invariant Phe residue (CD1) has been suggested to be important for supporting the heme. The present analysis revealed that this residue is also well suited to the site in terms of energy. PMID- 9350983 TI - Leptin interacts with glucagon-like peptide-1 neurons to reduce food intake and body weight in rodents. AB - The adipose tissue hormone, leptin, and the neuropeptide glucagon-like peptide-1 (7-36) amide (GLP-1) both reduce food intake and body weight in rodents. Using dual in situ hybridization, long isoform leptin receptor (OB-Rb) was localized to GLP-1 neurons originating in the nucleus of the solitary tract. ICV injection of the specific GLP-1 receptor antagonist, exendin(9-39), at the onset of dark phase, did not affect feeding in saline pre-treated controls, but blocked the reduction in food intake and body weight of leptin pre-treated rats. These findings suggest that GLP-1 neurons are a potential target for leptin in its control of feeding. PMID- 9350984 TI - Two-dimensional distribution of Gi2alpha in the plasma membrane: a critical evaluation by immunocytochemistry. AB - Caveolae have been postulated as a center for signal transduction, because many signaling molecules are concentrated in caveolin-rich fractions. We took Gi2alpha as an example and examined whether it is constitutively concentrated in caveolae. First, the behavior of caveolin and Gi2alpha in density-equilibrium ultracentrifugation was reexamined. By collecting fractions efficiently, caveolin and Gi2alpha were found to distribute differently. Secondly, by novel immunocytochemical methods it was found that the labeling density of Gi2alpha was 2.29 times higher in caveolae than in the non-caveolar plasma membrane. The results indicate that the concentration of Gi2alpha in caveolae is lower than deduced from most biochemical studies. PMID- 9350985 TI - A cytoplasmic peptide of the neurotrophin receptor p75NTR: induction of apoptosis and NMR determined helical conformation. AB - The neurotrophin receptor (NTR) and tumor necrosis factor receptor family of receptors regulate apoptotic cell death during development and in adult tissues [Beutler and van Huffel, Science 264 (1994) 667-668]. We have examined a fragment of p75NTR from the carboxyl terminus of the receptor and a variant form of this peptide via NMR techniques and in vitro assays for apoptotic activity. The wild type peptide induces apoptosis and adopts a helical conformation oriented parallel to the surface of lipid micelles, whereas the variant form adopts a non helical conformation in the presence of lipid and shows no activity. These experiments suggest a link between structure and function of the two peptides. PMID- 9350986 TI - Preliminary NMR studies of Thermus thermophilus ribosomal protein S19 overproduced in Escherichia coli. AB - The gene for the ribosomal protein S19 from Thermus thermophilus was cloned, sequenced and overexpressed in Escherichia coli. A simple procedure for isolating the recombinant protein was developed. Preliminary NMR studies revealed a high content of alpha-helical secondary structure in the protein. PMID- 9350987 TI - Opposite effects of cell differentiation and apoptosis on Ap3A/Ap4A ratio in human cell cultures. AB - The biological role of diadenosine oligophosphates (DAOP) remains obscure in spite of numerous attempts to solve this enigma. It is known that Ap3A contrary to Ap4A accumulates in human cultured cells treated with interferons (IFNs) alpha or gamma. Since IFNs are considered as antiproliferative regulators, we assumed that different cell status may be associated with varying intracellular levels of DAOP. Promyelocytic human cell line HL60 induced by phorbol ester (TPA) to differentiate to macrophage-like cells in culture exhibits a profound loss of proliferative potential. Here we have shown a 4-5-fold increase in Ap3A concentration in HL60 cells induced by TPA, similar to the effect of IFN, while the Ap4A concentration remained unchanged. On the contrary, in cells undergoing apoptosis induced by VP16, a topoisomerase II inhibitor, the Ap3A concentration considerably decreased, while the Ap4A concentration increased. These findings combined with earlier results suggest an involvement of the Ap3A/Ap4A ratio in signal transduction pathways controlling the cell status. PMID- 9350988 TI - Telethonin, a novel sarcomeric protein of heart and skeletal muscle. AB - In this paper we describe a novel 19 kDa sarcomeric protein named telethonin. The cDNA sequence discloses an open reading frame of 167 amino acids that does not resemble any known protein. Antibodies against a recombinant telethonin fragment were used for Western blot analysis, confirming the presence of this 19 kDa protein in heart and skeletal muscle and revealing an immunofluorescence pattern typical of sarcomeric proteins, overlapping myosin. The frequency of specific cDNA clones in different libraries indicates that the telethonin transcript is amongst the most abundant in skeletal muscle. In human, telethonin maps at 17q12, adjacent to the phenylethanolamine N-methyltransferase gene. PMID- 9350989 TI - Intracellular Ca2+ dependence of nitric oxide mediated enhancement of interleukin 8 secretion in human endothelial cells. AB - Nitric oxide (NO.) can induce transient [Ca2+] changes in endothelial cells not different from receptor mediated signalling. Whether this Ca2+ signal may provide a link with IL-8 secretion induced by NO. donors was investigated in human endothelial cells. Sodium nitroprusside (SNP) and S-nitroso-N-acetyl-DL penicillamine (SNAP) dose dependently increased IL-8 production in this cell type. Additive IL-8 secretion was found with TNFalpha. Buffering intracellular Ca2+ with MAPT/AM suppressed NO. induced [Ca2+]i changes and reduced subsequent IL-8 secretion. The additive effect of both NO. donors on TNFalpha induced IL-8 secretion was completely blocked in the presence of MAPT/AM. SKF 96365, which has been shown to block receptor mediated Ca2+ entry, and TMB-8, which blocks intracellular Ca2+ release, both inhibited IL-8 secretion, particularly when TNFalpha was used as a costimulator, indicating that [Ca2+]i changes are important components of IL-8 induction by NO.. PMID- 9350990 TI - Temporal regulation of in vitro import of precursor proteins into tobacco mitochondria. AB - Protein import into isolated tobacco mitochondria was investigated using mitochondria from leaves harvested at different times of the day and night. Efficient import was only detected with mitochondria isolated from leaves harvested during the dark period of the growth cycle, only low levels of import were detected from leaves harvested during the light period. However, this temporal difference seen in import did not appear to be circadian in nature. This implies that the protein import process in mitochondria isolated from leaves is not constitutive. This has important implications for targeting specificity studies performed in transgenic plants, as unless the plants are tested at the time when import is occurring, the true in vivo targeting abilities of chimeric constructs will not be measured. PMID- 9350991 TI - Inhibition of glucose-induced insulin secretion by long-term preexposure of pancreatic islets to leptin. AB - Here we evaluated the effect of leptin on glucose-induced insulin secretion by normal rat pancreatic islets. We show in perifusion experiments that leptin had no acute effect on the secretory activity of beta-cells. However, following preexposure to leptin a pronounced time- and dose-dependent inhibition of both first and second phases of secretion was observed. Maximum inhibition was obtained at 24 h and with 100 nM leptin. This inhibition did not involve a decrease in cellular insulin content. It was also not observed with islets from fa/fa rats. Leptin thus inhibits insulin secretion by a mechanism which requires long-term preexposure to the hormone and which may involve alteration in beta cell gene expression. PMID- 9350992 TI - Unfolding of dimeric creatine kinase in urea and guanidine hydrochloride as measured using small angle X-ray scattering with synchrotron radiation. AB - The denaturation of dimeric creatine kinase (CK) induced by urea and guanidine hydrochloride (GuHCl) has been studied by small angle X-ray scattering (SAXS), which is a direct way to measure the changes in the overall dimensions of a protein molecule. The radii of gyration (Rg) of CK are 29+/-0.4 angstroms in the native state and 46+/-1.5 angstroms in the unfolded state in either 8 M urea or 3 M GuHCl. The transition curves of urea denaturation derived from the Rg values and the zero angle intensity (I(0)) are similar to that from intrinsic fluorescence, indicating that the changes in the molecular shape, the tertiary structure and the dissociation of the subunits proceed simultaneously. In the case of GuHCl-induced denaturation, the dramatic increases both in Rg and in I(0) in 0.3-0.5 M GuHCl suggest clearly that soluble aggregates form at low GuHCl concentrations. The aggregates dissociate and the molecule unfolds at higher GuHCl concentrations. The results suggest that the mechanisms of CK denaturation in urea and in GuHCl are somewhat different and the intermediate in GuHCl denaturation can much more easily form soluble aggregates. PMID- 9350993 TI - Identification of the human Lewis(a) carbohydrate motif in a secretory peroxidase from a plant cell suspension culture (Vaccinium myrtillus L.). AB - This paper reports for the first time the presence of the human Lewis(a) type determinant in glycoproteins secreted by plant cells. A single glycopeptide was identified in the tryptic hydrolysis of the peroxidase VMPxC1 from Vaccinium myrtillus L. by HPLC/ESI-MS. The oligosaccharide structures were elucidated by ESI-MS-MS and by methylation analysis before and after removal of fucose by mild acid hydrolysis. The major structure determined is of the biantennary plant complex type containing the outer chain motif Lewis(a) [structure in text]. A corresponding fucosyltransferase activity catalyzing the formation of Lewis(a) type structures in vitro was identified in cellular extracts of the suspension cultures. PMID- 9350994 TI - Identification of positively charged residues contributing to the stability of plasminogen activator inhibitor 1. AB - Plasminogen activator inhibitor 1 (PAI-1), a member of the serpins, has a unique conformational flexibility. A typical characteristic is its intrinsic lability resulting in the conversion of the active conformation to a latent conformation. In the present study, we have evaluated the effect of substitution of positively charged residues located at the turn connecting strand s4C with strand s3C, either with negatively charged or with neutral residues, on the functional stability of PAI-1. The following mutants were constructed, purified and characterized in comparison to wild-type (wt) PAI-1: PAI-1-R186E,R187E (Arg186--> Glu and Arg187--> Glu), PAI-1-H190E,K191E (His190--> Glu and Lys191--> Glu) and PAI-1-H190L,K191L (His190--> Leu and Lys191--> Leu). In contrast to wtPAI-1 the mutants exhibited no inhibitory activity. Whereas latent wtPAI-1 can be reactivated (up to a specific activity of 78+/-19%) by treatment with guanidinium chloride, a similar treatment applied to these mutants resulted in a significant but relatively small increase of specific activity (i.e. to 14%). Evaluation of the functional stability (at 37 degrees C, pH 5.5, 1 M NaCl revealed a strongly decreased functional stability compared to wtPAI-1 (i.e. 3-9 h for the mutants vs. > 24 h for wtPAI-1). Further characterization by heat denaturation studies and plasmin susceptibility confirmed that removal or reversal of the positive charge on the turn connecting s4C with s3C results in PAI-1 mutants with a highly accelerated conversion of active to latent forms. We can therefore conclude that the pronounced positive charge in the turn connecting s4C with s3C is of the highest importance for the functional stability of PAI-1. PMID- 9350995 TI - Extensive cellular uptake into endothelial cells of an amphipathic beta-sheet forming peptide. AB - Extensive internalization into endothelial cells has been found for a water soluble amphipathic 26-mer beta-sheet peptide (FLUOS-DPKGDPKGVTVTVTVTVTGKGDPKPD NH2; VT5). With the D-Val13,D-Thr14 di-D-amino acid analog of VT5 (DD-VT5), exhibiting an identical primary structure but no propensity to adopt a beta-sheet conformation, only about 5% of the cellular uptake of VT5 was found. The mechanism of entry of VT5 into the cells remained unclear, but proved to be energy, temperature and pH dependent and, therefore, clearly distinct from that reported for helical amphipathic peptides. No detectable cytotoxicity, high solubility in water and the found extensive entry into endothelial cells make VT5 appear a good lead for developing new types of vectors for delivering oligonucleotides and peptides into intact cells. PMID- 9350996 TI - Agonist-induced signaling and trafficking of the mu-opioid receptor: role of serine and threonine residues in the third cytoplasmic loop and C-terminal domain. AB - The human mu-opioid receptor and a mutant form, muS/ T[i3+Cter]A, in which all Ser and Thr residues from the third cytoplasmic loop and C-terminal domain were changed to Ala, were studied after expression in CHO-K1 cells. Although the mutant receptors had similar affinities for agonists and EC50 values for inhibition of adenylyl cyclase as compared to wild-type receptors, the Emax were almost 2-fold decreased, suggesting a role of the mutated residues in G-protein coupling. After chronic morphine or etorphine, the EC50 values of the agonists were about 5-fold increased at both receptors but the Emax values were not altered; upon agonist withdrawal forskolin-stimulated cAMP levels were increased to almost 200% of control levels. Sequestration and rapid down-regulation of the mu-opioid receptor were induced by DAGO and etorphine but not morphine. In contrast, the muS/T[i3+Cter]A receptor was not sequestered and was up-regulated (150-380%) after treatment with agonists. The results indicate that the Ser and Thr residues in the third cytoplasmic loop and C-terminus of the mu-opioid receptor are not involved in the limited desensitization or in the adenylyl cyclase superactivation promoted by agonists but that their integrity and/or their phosphorylation is required in the intricate and coordinately regulated pathways involved in receptor signaling and trafficking. PMID- 9350997 TI - A new family of K+ transporters from Arabidopsis that are conserved across phyla. AB - Transport of K+ in higher plants, as in bacteria and fungi, is mediated by two broad classes of transport proteins that operate in the millimolar and micromolar K+ concentration ranges. A search of the Expressed Sequence Tag database using amino acid consensus sequences for the K+ transporters HAK1 from Schwanniomyces and Kup of Escherichia coli yielded two homologous sequences for Arabidopsis. Cloning and sequencing of these genes gave single open reading frames for the putative transporters, AtKT1 and AtKT2, with predicted molecular weights of 79 and 88 kDa. The predicted gene products showed a high degree of homology at the amino acid level (56% identity) and exhibited significant hydrophobic stretches in their N-terminal halves, consistent with 12 membrane-spanning, alpha-helical domains. Database searches using AtKT1 and AtKT2 identified 10 additional sequences in Arabidopsis as well as additional homologous sequences in the plant species Oryza and Allium, the bacterium Lactococcus lactis, and in Homo sapiens. Expression of AtKT2 rescued growth on low millimolar [K+] in Saccharomyces cerevisiae carrying deletions for the genes encoding the K+ transporters TRK1 and TRK2. Rescue was associated with a 2-fold stimulation of Rb+ uptake and was sensitive to competition with external Na+ but not to extracellular pH, indicating that the gene encodes a low-affinity K+ transporter. These and additional results suggest that AtKT1 and AtKT2 belong to a superfamily of cation transporters that have been conserved through evolution. PMID- 9350998 TI - Matrilin-3 from chicken cartilage. AB - By subtractive cDNA cloning we have identified a novel constituent of chicken cartilage termed matrilin-3. This constituent is encoded by a mRNA of 2.2 kbp whose expression is restricted to cartilaginous tissues. The predicted protein is composed of 452 amino acids with a molecular mass of 49 kDa. It contains a single von Willebrand factor A-like domain, four epidermal growth factor-like repeats and an alpha-helical region which may induce the formation of oligomers via a coiled-coil. The primary structure is similar to that of matrilin-1 which is also expressed in a cartilage-specific manner. This similarity suggests that the genes for the two proteins may have evolved from a common ancestor by gene duplication. PMID- 9350999 TI - Repression of apoC-III gene expression by TNFalpha involves C/EBPdelta/NF-IL6beta via an IL-1 independent pathway. AB - Apolipoproteins A-II and C-III, which participate in the control of cholesterolemia and triglyceridemia, are negative acute phase proteins. Treatment of HepG2 cells with TNFalpha showed that apoA-II and apoC-III mRNA levels were decreased. Using transient transfection, we found that apoC-III gene expression is controlled at the transcriptional level. By competition and supershift experiments, we demonstrate that TNFalpha-induced complexes were related to C/EBPdelta/NF-IL6beta and p50 and that overexpression of C/EBPdelta was able to reproduce the inhibitory effect of TNFalpha on the apoC-III promoter. RT-PCR failed to detect the IL-1 transcript in TNFalpha-treated HepG2 cells, suggesting that activation of C/EBPdelta by TNFalpha is not related to the IL-1-signalling pathway. PMID- 9351000 TI - Sequential assignments and secondary structure of the RNA-binding transcriptional regulator NusB. AB - The NusB protein is involved in transcriptional regulation in bacteriophage lambda. NusB binds to the RNA form of the nut site and along with N, NusA, NusE and NusG, stabilizes the RNA polymerase transcription complex and allows stable, persistent antitermination. NusB contains a 10 residue Arg-rich RNA-binding motif (ARM) at the N-terminus but is not sequentially homologous to any other proteins. In contrast to other known ARM-containing proteins, NusB forms a stable structure in solution in the absence of RNA. NMR spectroscopy was used to determine that NusB contains six alpha-helices: R10-Q21, 127-F34, V45-L65, Q79-S93, Y100-F114 and D118-L127. The structure of NusB makes it a member of a newly emerging class of alpha-helical RNA-binding proteins. PMID- 9351001 TI - Cleavage of Gag precursor is required for early replication phase of HIV-1. AB - A mutant of human immunodeficiency virus type 1 (HIV-1), which is deficient for Gag precursor cleavage and noninfectious, was characterized with respect to its defective step in the viral replication phase. Upon transfection, the mutant produced a normal level of progeny virions as monitored by electron microscopy and RNA hybridization. Single-round replication assay demonstrated, in contrast, that the mutant was defective at the early phase of the replication cycle. Furthermore, no viral DNA was detected in the cells infected with the mutant. Taken together, it is concluded that maturation of Gag precursor protein of HIV-1 is required for an early event(s) before or during a coupled process of uncoating/reverse transcription. PMID- 9351002 TI - HIV-1 capsid mutants inhibit the replication of wild-type virus at both early and late infection phases. AB - In-frame mutations were introduced into various portions of the human immunodeficiency virus type 1 (HIV-1) gag gene, and potentials of the mutants to suppress the replication of wild-type HIV-1 were monitored. In contrast to results obtained with matrix and nucleocapsid mutants, almost all capsid mutants blocked HIV-1 replication completely in single-round replication assays. A capsid mutant designated C6b was demonstrated to be one of the most efficient inhibitors for HIV-1 reported to date, and to be effective at both early and late viral replication phases. T-cells, which are engineered to express the C6b Gag in response to HIV-1 infection, were perfectly resistant to HIV-1. PMID- 9351003 TI - Improving scFv antibody expression levels in the plant cytosol. AB - Expression of single-chain antibody fragments (scFvs) in the plant cytosol is often cumbersome. It was unexpectedly shown that addition at the C-terminus of the ER retention signal KDEL resulted in significantly improved expression levels. In this report the cytosolic location of the scFv-CK was confirmed, excluding possible mistranslocation to other subcellular compartments. It was shown that expression of several other scFvs was also improved in tobacco protoplasts. In addition expression was improved in transgenic potato. Changing from KDEL to KDEI did not affect the enhanced protein expression level. Addition of the KDEL motif is a simple and straightforward tool to stabilize in planta cytosolic expression of many scFvs. PMID- 9351021 TI - Assignment of glial brain tumors in humans by in vivo 1H-magnetic resonance spectroscopy and multidimensional metabolic classification. AB - This study presents a simple approach for the noninvasive assignment of glial brain tumors according to malignancy by single-voxel proton magnetic resonance spectroscopy at short echo times (TE < or = 50 milliseconds). Based on peak area ratios, a five-dimensional data set was obtained for each investigated subject. This vector was then projected along metabolic coordinates in a two-dimensional metabolic space. These coordinates had been determined in a previous study (Hagberg G et al., 1995, Magn Reson Med 34: 242-252). Tumor assignment was done without any knowledge of histology by comparing the location of the new cases to the features of the previous study. All 11 investigated glioblastomas multiforme, as well as 4 of 5 astrocytomas grade II, could easily be assigned to the groups of high- and low-grade tumors, respectively. Classification was more difficult in the case of a cystic astrocytoma grade II and one astrocytoma grade III. Two spectra measured in normal-appearing matter of glioblastoma patients were not classified as healthy. Using single-voxel proton magnetic resonance spectroscopy at short echo times with the knowledge of a base study, a straightforward, fast, and noninvasive differential diagnosis of glial brain tumors is possible. PMID- 9351022 TI - Effects of riluzole on the evolution of focal cerebral ischemia: a magnetic resonance imaging study. AB - The aim of this study was to investigate the effects of riluzole on the lesion induced by a permanent middle cerebral artery occlusion (MCAO) in rats. Riluzole at 4 or 8 mg/kg i.v. significantly reduced the cortical ischemic brain damage. With the most effective dose of 8 mg/kg, the time evolution of the lesion was assessed by T2-weighted magnetic resonance imaging (MRI) repeated on the same animals after MCAO. MRI obtained at 24, 48, and 72 hours after MCAO showed a progressive increase of the ischemic lesion, except in the cortex of the riluzole treated rats (8 mg/kg i.v.). Furthermore, there was no difference between lesion volumes as measured by MRI or by histology. This study indicates that MRI may be a valuable method to quantify in vivo the neuroprotective profile of a drug. PMID- 9351023 TI - Contrast-modified gradient echo imaging using rotary echo preparatory pulses. AB - The use of on-resonance 121 binomial composite pulses in two- or three dimensional magnetization-prepared gradient-recalled echo magnetic resonance imaging experiments generates rotary echoes, leading to an increase in contrast range that is, in part, determined by the ratio of T2 to T1. In comparison with other fast gradient-recalled echo imaging techniques designed for enhanced T2 contrast, this method is more robust with respect to radiofrequency field inhomogeneity and less sensitive with respect to motion artifacts. Three dimensional parametric images may be calculated using least-squares fitting based on a simple model for steady-state longitudinal magnetization during the imaging sequences. PMID- 9351024 TI - 1H-spectroscopic imaging with read gradient during acquisition in inhomogeneous fields: analysis, measurement strategy, and data processing. AB - The proton magnetic resonance spectroscopic imaging techniques that use read gradient during acquisition produce proton spectra with high spatial and moderately high spectroscopic resolution in a reasonable time for in vivo applications. These techniques suffer mainly from the spatial and spectral distortions caused by the convolution of spectral/spatial information (chemical shift artifacts) and from the spectral shifts caused by static magnetic field inhomogeneities. The investigators analyze the chemical-shift artifacts in the presence of nonnegligible static magnetic field inhomogeneities and propose a postdetection processing scheme to correct for such effects. Spectral artifacts caused by chemical shifts, spectral line overlapping, streak broadening, and magnetic field inhomogeneities are discussed. The postdetection data processing scheme is demonstrated on measurements of a phantom as well as a human leg. PMID- 9351025 TI - Investigation of blood-brain barrier permeability to magnetite-dextran nanoparticles (MD3) after osmotic disruption in rats. AB - The permeability of experimentally disrupted blood-brain barrier (BBB) to superparamagnetic nanoparticles (MD3) was studied in rats. BBB opening was induced by intracarotid injection of mannitol. One hundred eighty rats were used for the study. Rats were examined at two time points, 30 minutes and 12 hours after intracarotid mannitol injection. Different preparations intravenously injected 30 minutes before rat sacrifice were used for characterization of BBB disruption. BBB integrity was determined with 99mTc-diethylenetriamine pentaacetic acid (DTPA) and 99mTc-albumin. Iron oxide-glucose particles (12-nm mean diameter), 99mTc-labeled lecithin-cholesterol liposomes of three different sizes (50, 100, and 200 nm), and polyethylene glycol (PEG)-coated 99mTc liposomes (50 nm) were used for investigations of the dependence of BBB permeability on particle system size or surface. Magnetite-dextran nanoparticles (MD3) were evaluated as superparamagnetic contrast agent to monitor with magnetic resonance imaging (MRI) the BBB breakdown. In vitro T1 and T2 relaxation times of the brain tissue were measured at 40 MHz and 37 degrees C, and T2-weighted MR images were acquired at 0.5 T. After intracarotid mannitol infusion, as expected, the BBB breakdown was immediate and temporary as judged by soluble molecule diffusion. MD3 nanoparticles crossed the BBB 12 hours after intravenous mannitol injection, at a time when brain permeability for molecules or small particles returns to normal. Magnetite crystals were found in cytoplasmic vesicles of glial cells. On MRI, signal intensity decreased after injection of MD3, even 12 hours after mannitol injection. This particularity could be useful in the study of focal pathological lesions accompanied by BBB permeability modifications. In such conditions, superparamagnetic particle contrast agents could be caught by the BBB, allowing the observation of impaired BBB areas without detectable cellular lesions. PMID- 9351026 TI - Changes in proton transverse relaxation times of rat myocardium that has suffered a previous oxidative insult. AB - An oxidative insult can induce severe damage, as in the phenomenon of myocardial ischemia and reperfusion. However, there are situations in which the damage is not so obvious (e.g., silent ischemia or aging), and the negative effects will be seen only in time. Our aim was to reveal these small changes in the myofilaments by using the nuclear magnetic resonance (NMR) technique. We used Wistar rat hearts in a constant-pressure Langendorff system, perfused with oxygenated Krebs Henseleit buffer at 37 degrees C. After 15 minutes of stabilization, the hearts were perfused with buffer supplemented with H2O2 at 50, 75, or 100 micromol/L for 15 or 30 minutes. Fifteen-minute and 45-minute perfusion controls and unperfused hearts were also collected. Heart rate (HR) and left ventricular developed pressure (LVDP) were determined with the help of a latex balloon, inserted in the left ventricle and connected with a pressure transducer. Proton transverse relaxation times (T2) were determined at the end of the experiment. T2 values were measured again in the same tissue fragments after they had been glycerinated and incubated in relaxation and rigor media. The functional parameters (HR, LVDP, coronary flow) were not significantly changed in control and 50 micromol/L H2O2 groups but were increased in the 75 micromol/L H2O2 group and significantly decreased in the 100 micromol/L H2O2 group. T2 is significantly decreased in rigor media starting with 50 micromol/L H2O2 administrated for 30 minutes and does not correlate with dose and duration of the oxidative insult. T2 in rigor is shorter than in relaxation media within the groups, and this difference is increased in the treated hearts. PMID- 9351028 TI - Double-tuned four-ring birdcage resonators for in vivo 31P-nuclear magnetic resonance spectroscopy at 11.75 T. AB - A high signal-to-noise ratio (SNR) in 31P-nuclear magnetic resonance (31P-NMR) spectroscopy can be obtained only with good B0-field homogeneity and optimal coil sensitivity. This demands double-tuned coils with a highly sensitive 31P channel and an additional 1H channel for 1H-magnetic resonance imaging, shimming, 1H decoupling, and nuclear Overhauser enhancement (NOE). For studies on an 11.75 T magnet, we built coils derived from the four-ring birdcage design originally described by Murphy-Boesch. A comparison with conventional, single-tuned coils shows that, in spite of double tuning, there is no significant loss in 31P sensitivity while the 1H channel provides the requested performance. The coil design offers the advantage of circular polarization on both channels. PMID- 9351027 TI - Physiological constraints on changes in pH and phosphorus metabolite concentrations in ischemically exercising muscle: implications for metabolic control and for the interpretation of 31P-magnetic resonance spectroscopic studies. AB - Relationships between pH and the concentrations of phosphocreatine (PCr), inorganic phosphate (Pi), and lactate during ischemic exercise depend on passive buffering, proton consumption as a consequence of net PCr breakdown, the control of glycogenolysis, (particularly in relation to the concentration of Pi, a substrate of glycogen phosphorylase that is produced by net PCr breakdown), and the creatine kinase equilibrium. The author analyzes the implications of these relationships for the interpretation of 31P-magnetic resonance spectroscopic data and for the control of glycogenolysis. For realistic adenosine diphosphate (ADP) concentrations, given the constraints of the creatine kinase equilibrium, the pH must be near-linear with lactate, with an apparent buffer capacity (i.e., the ratio of lactate accumulation to pH change) that is nearly twice the true buffer capacity (i.e., the ratio of net proton loading to pH change). The implications for glycogenolytic control depend on adenosine triphosphate (ATP) turnover, but an upper limit of activation of glycogen phosphorylase (i.e., the amount of the a form) that would permit no increase in ADP concentration can be calculated. Phosphorylase activation during ischemic exercise seems approximately proportional to the power output, consistent with calcium stimulation of phosphorylase b kinase. In simulations, ADP concentration is highly sensitive to this proportionality, as (unlike in purely oxidative exercise) ADP concentration is not known to participate in any closed feedback loops in ischemic exercise. PMID- 9351029 TI - Gadolinium-enhanced magnetic resonance angiography of the pelvis in patients with erectile impotence. AB - This study evaluated the potential of contrast-enhanced digital-subtraction magnetic resonance angiography (CE-DS-MRA) for noninvasive angiographic delineation of the arterial supply of the penis in patients with erectile dysfunction. After induction of an erection with prostaglandin E, a three dimensional fast imaging with steady-state precision (FISP) sequence with TE of 1.8-2 milliseconds, TR of 4.4-5 milliseconds, and flip angle of 40 degrees-60 degrees was used to obtain high-resolution angiograms of the pelvis and penis during the injection of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) 0.3 mmol/kg body weight, within 30-50 seconds. DS maximum intensity projections (MIPs) and multiplanar reconstructions (MPRs) were compared with clinical work-up and directional Doppler ultrasound in 11 patients. In all 11 patients (100%), the arterial supply of the penis could be delineated from the aortic bifurcation via the iliac and internal pudendal arteries to the dorsal and deep penile arteries. Of the 22 internal pudendal arteries, 6 (27%) were occluded on CE-DS-MRA and 5 (23%) had stenoses, of which 4 (18%) were greater than 50%. In 7 patients (64%) good correlation between CE-DS-MRA and clinical findings and/or Doppler ultrasound was found; in 2 patients (18%), the correlation was moderate, and in 2 patients (18%) results were discrepant. In 6 patients (55%), MRA provided additional information to the clinical and Doppler ultrasound work-up. CE-DS-MRA can delineate small vessels such as the internal pudendal and penile arteries and thus has the potential to become a noninvasive angiography method in the work-up of erectile impotence. PMID- 9351030 TI - Cure with omeprazole plus amoxicillin versus long-term ranitidine therapy in Helicobacter pylori-associated peptic ulcer bleeding. AB - BACKGROUND: Long-term prophylaxis with ranitidine reduces the risk of recurrent bleeding in patients with a history of bleeding peptic ulcers. To date, no randomized study has been performed to compare cure of Helicobacter pylori infection versus H2 blocker prophylaxis in patients with bleeding peptic ulcer. METHODS: In a prospective randomized study, 95 consecutive patients with H. pylori-associated peptic ulcer bleeding were randomized to either ranitidine prophylaxis (150 mg at night) for 2 years or to H. pylori-eradication with omeprazole 60 mg twice daily plus amoxicillin 750 mg three times daily for 10 days. RESULTS: (Intention-to-treat analysis). Forty-eight patients were enrolled in the ranitidine group; 47 in the omeprazole-plus-amoxicillin group. Mean follow up was 576 days (77 to 730). Ulcer recurrence rate was 31.3% in the ranitidine group (group 1) versus 6.37% in the eradication group (group 2; p = 0.0018). More patients had recurrent bleeding in group 1 (8.3%) than in group 2 (4.2%) but we were not able to show a statistically significant difference with respect to recurrent bleeding between groups (p = 0.29). Definite cure of H. pylori infection was achieved in 89.3%. CONCLUSIONS: Cure of H. pylori infection reduces recurrence of peptic ulcer and therefore rebleeding more effectively than does long-term maintenance therapy with an H2 blocker. PMID- 9351032 TI - Endoscopic submucosal tumorectomy for gastrointestinal submucosal tumors restricted to the submucosa: a new form of endoscopic minimal surgery. AB - BACKGROUND: Endoscopic minimally invasive therapy for submucosal tumors of the gastrointestinal tract by use of endoscopic ultrasound has not yet come into widespread use, and this technique has not been fully evaluated. We therefore investigated this method of treatment in patients with gastrointestinal submucosal tumors. METHODS: Forty-five patients with suspected gastrointestinal submucosal tumors (esophagus [5], stomach [1], duodenum [16], colon [23]) based on barium enema studies and endoscopy underwent endoscopic ultrasound by the water-filled or balloon method. The layer of origin and the internal echogenicity of the lesions were evaluated. After confirming that the tumors were submucosal, the lesions were resected using injection of physiological saline solution and electrocautery. RESULTS: Using a one-channel or two channel method, all tumors were completely resected without serious complications and the diagnosis was histologically confirmed. Ulceration at the site of resection healed within 2 to 4 weeks (mean 23 days) and there has been no local recurrence. CONCLUSIONS: Our technique of endoscopic submucosal tumorectomy appears to be a safe and useful diagnostic-cum-therapeutic procedure for gastrointestinal submucosal tumors. PMID- 9351031 TI - Endoscopy is not a risk factor for Helicobacter pylori infection--but medical practice is. AB - BACKGROUND: Previous studies have suggested an increased risk for Helicobacter pylori infection in physicians who perform UGI endoscopy because of exposure to potentially infectious gastric secretions. Therefore, the H. pylori infection status of the endoscopy staff was compared with the H. pylori prevalence of medical staff without endoscopy experience and control subjects who had no contact with patients. METHODS: The noninvasive 13C-urea breath test was performed in 2108 volunteers: 1460 physicians (mean age 44 +/- 12 years), 235 nurses (33 +/- 10 years), and 413 control subjects (43 +/- 12 years) who were not working in clinical medicine. All subjects completed a questionnaire concerning the weekly frequency of gastroscopies and the duration of endoscopic experience. RESULTS: Overall, 37.4% of the physicians and 35.3% of the nurses, but only 27.1% of the control subjects were infected. H. pylori infection was not significantly different between endoscopy-performing (37.8%; n = 1091) and general medical staff (35.9%; n = 604). Neither the frequency of gastroscopies nor the duration of endoscopy practice correlated with H. pylori status. With respect to the age distribution; however, a statistically significant higher prevalence of H. pylori was observed in physicians and nurses compared with the 413 control subjects without patient contact (p < 0.01). CONCLUSION: UGI endoscopy is not a risk factor for H. pylori infection, but medical practice slightly raises H. pylori acquisition. PMID- 9351033 TI - Usefulness of cytopathology and histology in the evaluation of Barrett's esophagus in a community hospital. AB - BACKGROUND: Brush cytology and histology have been found to be complementary in the evaluation of Barrett's esophagus at a referral medical center. This study evaluated the usefulness of brush cytology and histology in a community hospital setting. METHODS: One hundred consecutive patients with Barrett's esophagus underwent esophagogastroscopy performed by four staff gastroenterologists. Four quadrant biopsy specimens for histopathology at 3-cm intervals throughout the Barrett's segment and one brushing for cytology were obtained. All specimens were interpreted by four board-certified staff pathologists in a blinded fashion. RESULTS: Histologic diagnosis included three adenocarcinomas, one high-grade dysplasia, six low-grade dysplasias, and one indeterminate dysplasia. Cytology diagnosed the same three adenocarcinomas, no high-grade dysplasia, three low grade dysplasia, and two indeterminate dysplasias. The case of high-grade dysplasia on histology was diagnosed as normal by cytology. The six patients found to have low-grade dysplasia by histology were found to have low-grade dysplasia (3), indeterminate dysplasia (2), and no abnormality (1) by cytology. In no case was a higher grade of dysplasia diagnosed by cytology than by histology. CONCLUSION: Adding brush cytology to histology increased the cost but not the diagnostic yield in the evaluation of Barrett's esophagus in a community hospital setting. PMID- 9351034 TI - Prospective study of bacteremia rate after elastic band ligation and sclerotherapy of esophageal varices in patients with hepatosplenic schistosomiasis. AB - BACKGROUND: Esophageal band ligation is considered to be as efficient as endoscopic sclerotherapy, with a lower complication rate, including bacteremia. There are few studies comparing the two methods. The aim of this study was to compare the incidence of bacteremia after both treatments in patients with portal hypertension secondary to schistosomiasis. METHODS: Endoscopic sclerotherapy and band ligation were performed using standard techniques. Blood samples were obtained 5 and 30 minutes after endoscopic band ligation or sclerotherapy and cultured for aerobic and anaerobic organisms. RESULTS: In the sclerotherapy group 2 of 43 (4.6%) blood cultures were positive (Peptostreptococcus sp and Streptococcus mitis). A similar result was obtained in the band ligation group: 2 of 35 (5.7%) had positive cultures, both with Staphylococcus aureus. CONCLUSIONS: There is no difference in the frequency of bacteremia after treatment of esophageal varices with endoscopic sclerotherapy or endoscopic band ligation in patients with portal hypertension secondary to schistosomiasis. PMID- 9351035 TI - The uncleared fundal pool in acute upper gastrointestinal bleeding: implications and outcomes. AB - BACKGROUND: The implications and outcomes of patients with an uncleared fundal pool of blood found at emergent upper endoscopy are not well described. METHODS: We reviewed the records of 484 consecutive patients who presented over a 12-month period to our medical center with acute upper gastrointestinal hemorrhage. All patients underwent upper endoscopy within 24 hours of their initial presentation. Patients with an uncleared fundal pool of blood at initial endoscopy were included in this study, and their findings and outcomes were compared with a randomly selected subgroup of these same patients who did not have residual gastric blood. RESULTS: Sixty-one patients (13%) had uncleared fundal pools despite gastric lavage and patient positioning. Findings on initial endoscopy included esophageal varices in 29 (47%), gastric ulcer in 12 (20%), portal hypertensive gastropathy in 5 (8%), Mallory-Weiss tear in 5 (8%), duodenal ulcer in 5 (8%), gastric varices in 4 (7%), Dieulafoy's lesion in 2 (3%), and other in 7 (11%). Twelve of these 61 patients had multiple findings and 4 (7%) had no lesion identified. Thirty-two of the 61 patients (52%) had at least one follow-up endoscopy, with new fundal lesions identified in 13 (41%): portal hypertensive gastropathy in 8, gastric ulcer in 2, gastric varices in 2, and leiomyoma in 1. Of these 13 new findings, 5 (38%) were judged significant either by the presence of active bleeding or stigmata of recent hemorrhage. Of the 4 patients with no identifiable lesion on initial endoscopy, 3 had a follow-up endoscopy and 2 were found to have a significant new finding in the fundus. The control group had a statistically significant lower percentage of endoscopic findings related to portal hypertension. Recurrent bleeding during the index hospitalization occurred in 54% of the patients with uncleared fundal pools versus 11% of the control group (0 < 0.01). Length of stay, number of units of blood transfused, need for emergent surgery for bleeding, as well as overall and bleeding-related mortality were all significantly greater in the patients with the uncleared fundal pool than in the control patients. CONCLUSIONS: The inability to clear a fundal pool of blood at emergent upper endoscopy is associated with significant morbidity and mortality. Further, new fundal lesions can be identified in 41% of patients on follow-up examination, with many being clinically significant. These data support the importance of clearing a fundal pool in patients undergoing endoscopy for upper gastrointestinal bleeding. PMID- 9351036 TI - Endoscopic ultrasonography for the initial staging and follow-up in patients with low-grade gastric lymphoma of mucosa-associated lymphoid tissue treated medically. AB - BACKGROUND: Endoscopic ultrasonography is an appropriate procedure to assess the depth of tumoral infiltration in primary gastric lymphoma. The aims of the present study were to characterize the endoscopic ultrasonographic aspects of low grade gastric lymphoma of mucosa-associated lymphoid tissue and to determine the value of this procedure in medical treatment assessment. METHODS: Between 1991 and 1996, 15 patients with low-grade gastric lymphoma of mucosa-associated lymphoid tissue were treated with oral cyclophosphamide and/or anti-Helicobacter pylori treatment. Endoscopic ultrasonography was carried out at the time of the diagnosis in all patients, 8 of whom (4 in complete remission and 4 with a stable or progressive disease) had at least one endoscopic ultrasonography examination within the treatment period (median follow-up 17 months). RESULTS: The initial procedure showed an increased gastric wall thickness from 6 to 12 mm in 8 patients, equal to 5 mm in 5 patients, and normal in 2 patients. The thickening was predominantly of the mucosa alone and/or the submucosa but never extended beyond the muscularis propria. No lymph node was found. Gastric wall thickness returned to normal in the 4 patients in complete remission and remained thick in 3 of the 4 patients with a stable or progressive disease. Of these 3 patients, at least one set of biopsy samples, carried out during follow-up, showed the absence of lymphoma, but histology performed subsequently found evidence of disease. CONCLUSIONS: Endoscopic ultrasonography differentiates superficial from infiltrative types of gastric lymphoma of mucosa-associated lymphoid tissue, which may have a prognostic significance and confirms remission or persistence of the disease with medical treatment during follow-up. When the gastric wall remains thick, even if histology is negative, repeated biopsies should be performed to detect evolving disease or relapse. PMID- 9351037 TI - The use of endoscopic ultrasonography to reduce the cost of treating ampullary tumors. AB - BACKGROUND: Local excision of selected ampullary tumors may result in the same benefit as Whipple resection with less morbidity and mortality. The purpose of this study was to determine if endoscopic ultrasonography could aid in the selection of patients for local resection and to determine if there was a significant cost difference between the two surgical procedures. METHODS: In this retrospective study of 32 patients who underwent surgery for ampullary tumors, endoscopic ultrasonography staging was performed in 18 patients. Resected specimens were used to determine pathologic staging. Local disease was defined as stage T2N0 or less. Cost data were available for 20 patients. RESULTS: The sensitivity and specificity of endoscopic ultrasonography for differentiating local from advanced ampullary tumors were both 83%. The median total cost for a local resection was $9314 versus $16,017 for a Whipple resection (p < 0.0017). CONCLUSION: Endoscopic ultrasonography is an effective tool for identifying patients with localized ampullary tumors. The cost of a local resection for ampullary tumors is significantly less than that of a Whipple resection. The use of endoscopic ultrasonography to select patients for local resection may be a cost-effective technique in the management of patients with ampullary tumors. PMID- 9351038 TI - The diagnostic yield of lower endoscopy plus biopsy in nonbloody diarrhea. AB - BACKGROUND: Patients presenting with diarrhea frequently undergo lower endoscopy plus biopsy as part of their diagnostic evaluation. The diagnostic yield of this approach has not been systematically evaluated. METHODS: To evaluate the diagnostic yield of endoscopy and biopsy in the investigation of nonbloody diarrhea, we performed a retrospective analysis using the endoscopy unit database of a tertiary care university hospital over a 3-year period. The database was searched for cases in which colonoscopy was performed for the single indication of diarrhea. The endoscopic findings and initial biopsy reports were extracted from a chart review, and each clinician was interviewed for the patient's current clinical diagnosis. The clinical diagnoses were compared with the endoscopy and biopsy results to determine whether the tests had contributed to making the clinical diagnoses. RESULTS: Three hundred six patients were identified. One hundred one were excluded for standardized predefined exclusion criteria, leaving 205 evaluable patients, of whom 77 had flexible sigmoidoscopy and 128 had colonoscopy. Eighteen percent had specific clinical diagnoses facilitated by endoscopy and/or biopsy. Endoscopy and biopsy results were normal in 74% of cases. In 8% of the cases either the endoscopy or biopsy findings were inconsistent with the final clinical diagnoses. CONCLUSIONS: Endoscopy and biopsy are important diagnostic tools in the evaluation of patients with nonbloody diarrhea, leading to a specific diagnosis in nearly one fifth of cases. PMID- 9351039 TI - Lack of complications following short-term stent therapy for extrahepatic bile duct strictures in primary sclerosing cholangitis. AB - BACKGROUND: In 10% to 20% of patients with primary sclerosing cholangitis, a dominant stricture of an extrahepatic bile duct is responsible for symptoms and an exacerbation of cholestasis. The complications of a dominant stricture can usually be relieved by endoscopic placement of a stent through the stricture. The conventional policy of leaving stents in situ for 2 to 3 months is associated with a high incidence (e.g., 50%) of clinical deterioration due to stent occlusion. We have attempted to overcome this problem by substantially reducing the duration of stent placement. METHODS: Sixteen patients with symptomatic primary sclerosing cholangitis and dominant extrahepatic bile duct strictures were treated by stent placement for a median interval of only 9 days. RESULTS: In all patients endoscopic stent therapy was technically successful with a 7% incidence of transient procedure-related complications. During median follow-up of 19 months (range 7 to 27 months) serum biochemical evidence of cholestasis decreased substantially and 13 (81%) of the 16 patients became asymptomatic. No patient had a recurrence or exacerbation of either symptoms or biochemical evidence of cholestasis that could be attributed to stent occlusion. CONCLUSIONS: Short-term endoscopic stent therapy is a safe and effective treatment for symptomatic dominant extrahepatic bile duct strictures in patients with primary sclerosing cholangitis. PMID- 9351040 TI - High-intensity focused ultrasound transducers suitable for endoscopy: feasibility study in rabbits. PMID- 9351041 TI - Expandable thermal-shaped memory metal esophageal stent: experiences with a new nitinol stent in 129 patients. PMID- 9351042 TI - Endoscopic fibrin sealant injection: a novel method of closing a refractory gastrocutaneous fistula. PMID- 9351043 TI - Endoscopic management of a persistent pancreatopleural fistula. PMID- 9351044 TI - Mirizzi syndrome successfully treated by extracorporeal shock wave lithotripsy following endoscopic sphincterotomy. PMID- 9351045 TI - Magnetic resonance cholangiopancreatography: a problem solving modality. PMID- 9351046 TI - Effective palliation of a colovaginal fistula using a self-expanding metal stent. PMID- 9351047 TI - Massive hematochezia from a visible vessel within a stercoral ulcer: effective endoscopic therapy. PMID- 9351048 TI - Bleeding ulcers and Helicobacter pylori. PMID- 9351049 TI - Outcomes research in endoscopy: current status and future directions. PMID- 9351050 TI - Regression of gastric lymphoma of mucosa-associated lymphoid tissue by Helicobacter pylori eradication: endoscopic observation. PMID- 9351052 TI - Efficacy of EUS. PMID- 9351051 TI - Endoscopic balloon dilation and Helicobacter pylori eradication in the treatment of gastric outlet obstruction. PMID- 9351053 TI - Rectal examination during endoscopy: feel what you can't see. PMID- 9351054 TI - A novel approach to common bile duct stone extraction. PMID- 9351055 TI - Prospective randomized trial of endoscopic sclerotherapy versus variceal band ligation for esophageal varices: influence on gastropathy, gastric varices and variceal recurrence. PMID- 9351056 TI - Rapid measurement of urinary trypsinogen-2 as a screening test for acute pancreatitis. PMID- 9351057 TI - Benefit of endoscopic retrograde cholangiopancreatography in acute biliary pancreatitis-contemporary state at three surgery clinics in Prague. PMID- 9351058 TI - Acute pancreatitis-etiology, development, diagnosis and treatment. PMID- 9351059 TI - Endo-loop in the prevention of the post-polypectomy bleeding: preliminary results. PMID- 9351060 TI - What have we learned from the HOT (Hypertension Optimal Treatment) study? PMID- 9351061 TI - Compressive HLA matching. PMID- 9351062 TI - Molecular mechanisms of immunosuppressive chemical agents recently introduced in clinical transplantation protocols. PMID- 9351063 TI - Cyclosporin nephrotoxicity following cardiac transplantation. AB - The greatest change in GFR in response to treatment with cyclosporin occurs in the first 3-6 months and the magnitude of the decrement in the first year (or perhaps the first few months) appears to be a vital indicator of future problems. However, the apparent stabilization of renal function, particularly when monitored only by plasma creatinine, can conceal progressive tubulointerstitial injury, and increasing proteinuria is an ominous sign. Although lower doses of cyclosporin and careful monitoring of renal function may be helpful, there is at present no pharmacological intervention to protect or reverse the reduction in GFR that occurs. We believe that the vascular lesion induced by cyclosporin is fundamental, with early and initially reversible cyclosporin-induced vasospasm leading to progressive vascular damage with activation of endothelial cells and increased platelet interactions. Amongst other determinants, the renal response to this vasculopathy will depend on the balance between the presence of vasoactive factors with the vasoconstrictors promoting interstitial fibrosis and the vasodilators inhibiting proliferation. It is likely that the kidneys of heart transplant recipients are chronically ischaemic and as a consequence their renin angiotensin systems massively activated, which may further sensitize their kidneys to cyclosporin. Overproduction of angiotensin II, associated with the DD ACE genotype, has already been associated with poor prognosis in diabetic and IgA nephropathy. It is interesting to speculate that this ACE genotype, which is associated with a poor outcome in non-ischaemic heart disease can influence renal sensitivity to cyclosporin and predict the development of morphological injury. Extension of these experimental findings into the clinical arena with a placebo controlled trial of early introduction of ACE inhibitor therapy in recipients of cardiac transplants would be timely. PMID- 9351064 TI - Treatment of lupus nephritis--the advantages of a flexible approach. PMID- 9351065 TI - National Kidney Foundation: Dialysis Outcome Quality Initiative--development of methodology for clinical practice guidelines. AB - BACKGROUND: The NKF-DOQI (National Kidney Foundation--Dialysis Outcomes Quality Initiative) began in March 1995 with the following objectives: To improve end stage renal disease (ESRF) patient survival, to reduce patient morbidity, to increase efficiency of care, and to improve quality of life for ESRF patients. METHODS: The development of the NKF-DOQI Clinical Practice Guidelines focused on haemodialysis adequacy, peritoneal dialysis adequacy, vascular access, and anaemia. Principles guiding the methodology included scientific and methodological rigour, interdisciplinary approach, independence of the Work Groups, and an open review process. Unique features of this evidence-based guideline development process included the use of mock guidelines and rationales to guide the literature research, and an unprecedented three-stage review process. RESULTS: Work Groups reviewed more than 11,000 articles. A total of 114 clinical practice guidelines were developed by the Work Groups; these have been reviewed by more than 1200 professionals and patients. CONCLUSION: The NKF-DOQI Clinical Practice Guidelines has been a unifying effort for the entire renal community. The guidelines are an important step in the process of improving the quality of dialysis practice and improving ESRF patient outcomes. Priorities for implementation of the guidelines include education of ESRF professionals and patients, working with providers and insurers to encourage compliance, and follow up evaluation of patient outcomes to quantify compliance results. PMID- 9351066 TI - Inhibitory effect of Tripterygium wilfordii multiglycoside on increased glomerular albumin permeability in vitro. AB - BACKGROUND: Tripterygium wilfordii Hook F is a medicinal plant used for the treatment of glomerulonephritis in China. We studied the effect of Tripterygium wilfordii multiglycoside (TWG) on glomerular albumin permeability (Palbumin) in vitro. METHODS: Isolated rat glomeruli were incubated with protamine (600 micrograms/ml) for 30 min, or with human recombinant tumour necrosis factor (TNF alpha 0.4 ng/ml), superoxide (10 units/ml), or serum from a focal segmental glomerular sclerosis (FSGS) patient for 10 min at 37 degrees C. TWG, 1 mg/ml, was added in parallel tubes to study the effect on Palbumin. Control glomeruli were incubated under identical conditions. The albumin reflection coefficient (sigma albumin) was calculated from the change in glomerular volume in response to an applied oncotic gradient. Convectional permeability (Palbumin) was calculated as (1 - sigma albumin). RESULTS: Compared with controls, protamine increased the Palbumin of glomeruli (0.83 +/- 0.05, n = 25, vs 0.18 +/- 0.03, n = 20); pretreatment with TWG blocked this effect (0.13 +/- 0.04, n = 25). TNF-alpha also increased the Palbumin (0.79 +/- 0.04, n = 24 vs 0.04 +/- 0.07, n = 19); preincubation with TWG blocked this effect (0.03 +/- 0.09, n = 24). Palbumin of glomeruli incubated with xanthine and xanthine oxidase, resulting in the production of superoxide, also increased as compared to controls (0.85 +/- 0.04, n = 15 vs 0.08 +/- 0.05, n = 14); TWG blocked this effect as well (0.21 +/- 0.08, n = 14). FSGS serum also increased Palbumin of glomeruli significantly (0.88 +/- 0.02, n = 49 vs 0.00 +/- 0.02, n = 49); preincubation with TWG blocked this effect (0.05 +/- 0.07, n = 30). TWG by itself had no effect on Palbumin (0.19 +/- 0.10, n = 15). CONCLUSIONS: Our results show that TWG blocks protamine, TNF alpha, superoxide, and FSGS serum-mediated increase in glomerular albumin permeability in vitro. We conclude that reduction of proteinuria by Tripterygium wilfordii multiglycoside in various kinds of glomerular diseases in vivo might be due to protection of the glomerular filtration barrier. PMID- 9351067 TI - Frequency and impact of autosomal dominant polycystic kidney disease in the Seychelles (Indian Ocean). AB - BACKGROUND: As little such data is available in African populations, we investigated the prevalence of ADPKD and the impact of the disease in the Seychelles islands, where approximately 65% of the population is of African descent and 30% of Caucasian or mixed descent. METHODS: Prevalent cases were identified over a 3-year period by requesting all the doctors in the country (most of them are employed within a national health system) to refer all presumed or confirmed cases and by systematically examining the family members of all confirmed cases. The diagnosis was based on standard criteria including ultrasonographic findings and family history. RESULTS: Forty-two cases were identified in this population of 74,331 inhabitants, a total prevalence (per 100,000 total population) of 57 (95% CI, 41-76). All but one of the cases were of Caucasian descent so that the prevalence rates of the disease in the populations of Black and Caucasian descents were respectively 2 (0-11) and 184 (132-249). The prevalence rates of the gene(s) carriers were estimated to be 75 (45-117) in the total population respectively 6 (0-33) and 236 (140-372) in the Black and Caucasian populations. Haplotype analysis in 58 cases from three families showed a common DNA fragment in all affected individuals. Cases had significantly higher blood pressure compared to the general population and 21% had serum creatinine higher than 120 mumol/l. Among the established pedigrees, mean age of death between 1960 and 1995 (haemodialysis was introduced in 1992) was younger in subjects with than those without ADPKD (50.5 vs 67.7 years; P < 0.001). CONCLUSIONS: In the Seychelles, ADPKD clusters in the Caucasian population (possibly a founder effect), is rare in individuals of black descent, and is associated with substantial clinical and survival impact. PMID- 9351068 TI - Ambulatory blood pressure in hypertensive patients with autosomal dominant polycystic kidney disease. AB - BACKGROUND: Ambulatory blood pressure is more closely correlated with various indices of hypertensive target-organ damage, and is a better prognostic predictor of cardiovascular morbidity and mortality than conventional methods of blood pressure measurement. Autosomal dominant polycystic kidney disease (ADPKD) is complicated by hypertension, progressive renal failure, and an increased risk of cardiovascular mortality. This study investigated the 24-h ambulatory blood pressure profile in patients with ADPKD in view of the sparsity of such data in these patients and the possibility that abnormal diurnal blood pressure variations may have prognostic consequences. METHODS: Ambulatory blood pressure was measured over a 24-h period by the oscillometric method with an automatic non invasive recorder (SpaceLabs 90207 system) in matched groups of 25 hypertensive patients with ADPKD and 25 patients with essential hypertension. RESULTS: Both groups showed a nocturnal decrease in blood pressure, but this was significantly smaller in patients with ADPKD. There was no evidence of enhanced lability of blood pressure in ADPKD. CONCLUSIONS: The nocturnal fall in blood pressure was attenuated in patients with ADPKD. Further studies are required to assess the importance of this finding and its possible contribution to the progression of renal failure or increased cardiovascular mortality in these patients. PMID- 9351069 TI - Captopril-enhanced scintigraphy using the method of the expected renogram: improved detection of patients with renin-dependent hypertension due to functionally significant renal artery stenosis. AB - PURPOSE AND DESIGN OF STUDY: Asymmetric-induced changes of the renogram under angiotensin-converting enzyme inhibition (ACE-i), i.e. lateralization, is probably the most distinctive finding for the detection of haemodynamically significant renal artery stenosis (RAS) in compensated kidney, since bilateral and symmetric patterns are non-specific. In the Consensus statement of diagnostic criteria of renovascular hypertension with captopril renography (Am J Hypertens 1991; 4: 749-755S) ACE-i-induced asymmetry of renograms for the left and right kidney was viewed as vitally important. However, detection of change in split function is a reliable parameter only when using a glomerular tracer, i.e. 99mTc DTPA. No indication regarding a more widely used tubular tracer such as 99mTc mercaptoacetyltriglycine (99mTc-MAG3) has been given. METHODS AND RESULTS: The theoretical contralateral curve, called 'expected renogram', was calculated frame by frame from renal curves obtained under ACE-i and one of two baseline curves. The expected renogram was compared with the recorded ipsilateral curve. More than +/- 2 SD difference between expected and recorded renograms was assumed as suggestive of monolateral or bilateral RAS. Twenty-nine patients with angiographically proven RAS (bilateral in 12) and 20 patients without arteriographic evidence of stenosis were evaluated by postcaptopril/baseline 99mTc-MAG3 renography. Results obtained with the expected renogram analysis were compared with those obtained by standard criteria which included: improvement of peak time under baseline conditions, wash-out (75%) time, and monolateral or bilateral residual cortical activity > 10%, but asymmetrical, i.e. with > 5% change in split function. Compared to the standard evaluation, the use of the expected renogram for the diagnosis of RAS improved the specificity from 70 to 95% (P < 0.03) without loss of sensitivity (79.3%). Follow-up data after revascularization were available in 18 scintigraphically positive and six scintigraphically negative patients with RAS. The sensitivity of the expected renogram method referring to short-term (1 month) patient outcome following revascularization was 88.8%. The beneficial effects on blood pressure response persisted in 77% of the these patients at 18 months. Notably, four of six scintigraphically negative patients with RAS did not show any short-term benefit from revascularization and the improvement in blood pressure values lasted for 18 months in only one case. CONCLUSIONS: The high specificity of the expected renogram method reduces the number of unnecessary invasive procedures. This is a critical point for a low-prevalence disease such as renovascular hypertension. PMID- 9351070 TI - Energy metabolism in exertional heat stroke with acute renal failure. AB - BACKGROUND: Heat stroke is the clinical syndrome produced when the body overheats. It can develop in the army and in healthy civilian populations who physically exert themselves in a hot and humid environment during the summer, and may result in a significant number of heat-related deaths. Since strenuous exercise is one of the major exacerbating and precipitating factors, the incidence of exertional heat stroke (ExHS) is high among military personnel undergoing military training. Furthermore, acute renal failure (ARF) may occur in 25% of patients with ExHS and it can cause metabolic alterations that affect amino acid, carbohydrate, and lipid metabolism. Adequate nutritional support is essential for the treatment of ARF. The most important determinant of nutrient requirement in ARF is the degree of hypercatabolism caused by disease associated with renal function impairment. Indirect calorimetry (IDCM) is the method by which metabolic rate is estimated from measurements of oxygen consumption and carbon dioxide production. It can also provide information about the type and rate of substrate utilization in vivo (protein, carbohydrate, and fat). METHOD: The present clinical study is a comprehensive analysis of metabolic changes which includes energy expenditure (EE) and substrate utilization in 10 patients with ExHS with ARF and 10 patients with exertional heat exhaustion (ExHE) by the use of IDCM. RESULTS: Serum urea nitrogen, creatinine, peak creatine phosphokinase levels and heart rate were significantly increased in ExHS patients during ARF stage. Serum albumin levels were significantly decreased in ExHS patients with ARF. Resting energy expenditure (REE) was increased in patients with ExHS induced ARF and was not correlated with body temperature (r = 0.421). The average increase in EE during ARF stage was about 24%. The respiratory quotient in patients with ExHS induced ARF was lower than that in normal subjects and also in patients with ExHE. Urea nitrogen appearance rate increased in patients with ExHS induced ARF and in patients with ExHE without ARF. The percentage of total REE derived from fat in ExHS induced ARF and ExHE increased, while in patients with ExHS induced ARF and ExHE, the percentages of total REE derived from carbohydrate and protein were lower than those in control subjects. CONCLUSIONS: The present results suggest that patients with exertional heat injury (both ExHS and ExHE) have hypermetabolism during the acute stage. Furthermore, patients with exertional heat-induced rhabdomyolysis and ARF have a moderately higher hypermetabolism than those without ARF during the acute stage. We believe that this mainly reflects a more pronounced reduction of the vital cell mass (muscle) in relation to body weight, and/or a compromised substrate oxidation in ExHS with ARF. Whether or not this subgroup of patients will require a higher energy/caloric support merits further investigation. PMID- 9351071 TI - Odour perception in chronic renal disease. AB - BACKGROUND: The sense of smell plays an important role in the quality of life. Many studies have shown a declining odour perception in the elderly, as well as in subjects in poor health or nutritional state. Considering the high prevalence of poor nutritional state in renal disease and the importance of odour perception in nutrition and health, the relationship between renal function, nutritional state, and odour perception is explored in this study. METHODS: A total of 101 patients with chronic renal failure participated in the study. Thirty-eight haemodialysis patients (mean age = 64.3 years) were evaluated both before and after dialysis. Sixteen patients on peritoneal dialysis treatment (mean age = 64.0 years), 28 transplanted patients (mean age = 53.5 years, mean creatinine clearance = 64.0 ml/min) and 19 patients with varying degrees of renal insufficiency were also included (mean age = 63.7 years, mean creatinine clearance = 29.5 ml/min). Patients with cognitive deficits or upper respiratory airway diseases were excluded. A validated objective procedure was used to measure odour perception, by determining the detection threshold for isoamyl acetate (banana odour) as the lowest detectable odour concentration. RESULTS: Healthy control persons had significantly lower odour thresholds compared to patients on peritoneal (P = 0.001) and haemodialysis (P = 0.002). No significant difference was observed in odour perception between patients on peritoneal and haemodialysis (P = 0.779) and for patients on haemodialysis before and after a dialysis session. Transplanted patients had significantly better odour perception compared to matched patients on dialysis (P < 0.001). Odour perception of transplanted patients and matched healthy control persons was similar (P = 0.81). In patients with varying degrees of renal insufficiency, including healthy controls and transplanted patients, a significant positive correlation was found between odour perception and creatinine clearance (P = 0.02). A significant negative correlation was found between odour perception and serum concentration of urea (P < 0.001), serum phosphorus (P = 0.022) and protein catabolic rate (P < 0.05). Other parameters measuring nutritional status (albumin, BMI) were not correlated with odour perception. CONCLUSION: Our results show that the ability to smell is severely impaired in patients with chronic renal failure and is related to the degree of renal impairment and the degree of accumulation of uraemic toxins. After renal transplantation, patients have a normal odour perception, indicating the capacity of the olfactory system to recover once the concentration of uraemic toxins remains below a critical threshold. Acute removal of uraemic toxins by dialysis does not correct olfactory disturbances, suggesting a long lasting effect of uraemia on olfactory function. PMID- 9351072 TI - Serological evidence for reactivation of EBV infection due to uraemic immunodeficiency. AB - BACKGROUND: Reactivation of EBV infection is a common finding in immunocompromised individuals. The influence of 'uraemic immunodeficiency' on EBV infection is so far not well defined. METHODS: We determined specific antibodies to EBV nuclear antigens (EBNA) 1 and 2 in sera of 286 patients with immunodeficiency due to progressive chronic renal failure and of 51 healthy controls. We used the baculovirus vector expression system for recombinant production of EBNA1 and EBNA2. RESULTS: Serological evidence of reactivated or chronic persistent EBV infection, i.e. an anti-EBNA1/anti-EBNA2 ratio (E1/E2) < 1, was found in 18% of patients with chronic renal failure not yet receiving renal replacement therapy (CRF), 11% of peritoneal dialysis patients (CAPD), 25% of haemodialysis patients (HD), 24% of renal transplant recipients (TX), and in 6% of healthy controls. Rate of EBV reactivation was significantly increased in HD (P = 0.004) and TX (P = 0.006) patients compared to healthy controls. Moreover, the difference between HD and CAPD patients was statistically significant (P < 0.05). This finding may reflect additional effects modulating the function of the immunosystem, probably through activation of immunologically competent cells by contact with the artificial surfaces of dialysis membranes. Although the rate of EBV reactivations is expected to increase further under conditions of therapeutic immunosuppression, our serological approach did not detect an additional effect of immunosuppressive therapy following renal transplantation. However, this finding may reflect an impaired endogenous synthesis of antibodies caused by immunosuppressive agents. CONCLUSIONS: We conclude that determination of E1/E2 is useful for assessment of EBV infection in patients with chronic renal failure and 'uraemic immunodeficiency'. In patients with immunosuppressive therapy following renal transplantation additional testing including direct estimation of viral load, is necessary to correctly assess the state of EBV infection. PMID- 9351073 TI - Poor pre-dialysis glycaemic control is a predictor of mortality in type II diabetic patients on maintenance haemodialysis. AB - BACKGROUND: In type II diabetic patients, a better glycaemic control has been reported to slow down the progression of nephropathy. The effect of pre-dialysis glycaemic control on the long term prognosis in type II diabetics on haemodialysis is still uncertain. The purpose of this study is to evaluate the effect of glycaemic control before starting maintenance haemodialysis on the clinical outcome in type II diabetic haemodialysis patients. METHODS: One hundred and thirty-seven type II diabetics receiving regular haemodialysis in a single university hospital were enrolled. The patients were classified as either good or poor glycaemic control group according to their glycaemic control within 6 months before starting haemodialysis. Serum albumin, haematocrit, cholesterol, triglyceride, residual renal function, diabetic complications, and patient survival were analysed in both groups. RESULTS: There was no significant difference in age, gender, predialysis albumin level, cholesterol level, triglyceride level, and residual renal function between the two groups. The 1 year (94.5% vs 80.0%), 3-year (82.9% vs 58.1%), and 5-year (75.8% vs 21.8%) cumulative survival rates were lower in the poor glycaemic control group than in the good glycaemic control group (P < 0.001). The poor glycaemic control group also had more cardiovascular morbidity during the period of dialysis (P < 0.001). The increase in cardiovascular complications also accounted for the increased mortality during the course of haemodialysis. CONCLUSIONS: We conclude that poor glycaemic control before starting dialysis is a strong predictor of cardiovascular morbidity and survival for type II diabetics on haemodialysis. These results imply that better glycaemic control before dialysis might be important in improving the long-term prognosis in type II diabetics on haemodialysis. PMID- 9351074 TI - Time of diagnosis of chronic renal failure and assessment of quality of life in haemodialysis patients. AB - BACKGROUND: Time of diagnosis of chronic renal failure ani predialysis care may be important factors related to the quality of life of patients on dialysis treatment. METHODS: We evaluated the quality of life of 113 haemodialysis patients who had a late (< or = 1 month before starting dialysis, n = 53) or early (> or = 6 months, n = 60) diagnosis of chronic renal failure. At the time of the survey patients had been on dialysis for a median duration of 55 days (range 1-109). Quality of life was measured by the Kidney Disease Questionnaire (KDQ), including five dimensions with scales ranging from 1.0 to 7.0 (1.0 = more impairment): the health and life satisfaction indices (higher score = more dissatisfied), functional status (Karnofsky scale), and the time trade-off technique. RESULTS: Mean scores of quality of life measures were worse in the late- than in the early-diagnosis group. A significant difference (P < 0.05) was observed in the depression (4.46 +/- 1.45 vs 5.23 +/- 1.36), relationships with others (3.95 +/- 1.31 vs 4.53 +/- 1.31) and frustration (4.08 +/- 1.51 vs 5.21 +/ 1.34) dimensions of the KDQ. and in life satisfaction (4.11 +/- 1.92 vs 3.32 +/- 1.57). Functional status declined compared to 1 year before dialysis, particularly in the late-diagnosis group. Among the elderly patients, the magnitude of the difference was more pronounced, (including in the physical symptoms item of the KDQ). CONCLUSIONS: Our findings demonstrate that late diagnosis of chronic renal failure and the consequent lack of predialysis care adversely affect the quality of life of haemodialysis patients. Early diagnosis and regular predialysis care should be encouraged to improve the quality of life during dialysis treatment. PMID- 9351075 TI - Predicting future trends in the number of patients on renal replacement therapy in Denmark. AB - OBJECTIVES: To predict the future prevalence of patients on renal replacement therapy due to chronic renal failure in Denmark. SUBJECTS AND METHODS: Four thousand and nine terminal uraemic patients (median age 50.0 years, 15.2% diabetic) were treated in Denmark with renal replacement therapy in the period 1 January 1991 to 31 December 1995. Incidence rates and rates of transition between the treatment modalities (haemodialysis, peritoneal dialysis, and renal transplantation) were calculated. The prediction was made using a Markov model in three ways: (1) using the average rates (deterministic model), (2) using rates simulated with pseudorandom numbers based on the average rates (stochastic model), and (3) using increasing incidence rates in a deterministic model. RESULTS: Using present rates both model types predicted a significant increase in the prevalence of renal transplant recipients < 60 years (from 1003 in 1995 to about 1465 in 2006) and the prevalence of haemodialysis patients > or = 60 years (from 456 in 1995 to about 903 in 2006) while the prevalence of other treatment modalities would change less dramatically. The overall prevalence proportion would increase from 539 patients per million population (p.m.p.) in 1995 to about 777 p.m.p. in 2006. The stochastic model clearly demonstrated the uncertainties linked to the prognosis in contrast to the deterministic model. The deterministic model with increasing rates predicted a prevalence proportion of 1162 p.m.p. in 2006. CONCLUSION: Even with present rates the prevalence of haemodialysis patients in Denmark will continue to increase. Mathematical models offers a good tool to study future trends and to plan future capacity. PMID- 9351076 TI - Another simpler bypassing dialysate technique for measuring post-haemodialysis BUN. AB - BACKGROUND: The most popular method for estimating post-haemodialysis BUN is the low blood flow technique. The low blood flow technique with a blood flow of 50 ml/min for 3 min may encompass the first two phases of post-haemodialysis urea rebound by access and cardiopulmonary recirculations. Some patients may have risk of clotting in extracorporeal circuit while slowing blood flow. Therefore we proposed another simpler sampling technique for measuring post-haemodialysis BUN(C2). METHODS: In the present study, 28 long-term haemodialysis patients were divided into two groups. In group 1 (n = 15), C2 sample (C2-50) was collected immediately after blood flow was slowed down to 50 ml/min for another 3 min. Then blood flow was reset to 300 ml/min. Finally, C2 sample (C2B) was obtained from arterial port at the end of bypassing dialysate for another 3 min. In group 2 (n = 13), C2 sample (C2B) was obtained from arterial port at the end of bypassing dialysate for 3 min with a blood flow of 250 ml/min. Then dialysate was reset to original flow rate and C2 sample (C2-50) was collected soon after blood flow was slowed down to 50 ml/min for another 3 min. In the meantime, recirculation rate was also checked. RESULTS: The above two groups have similar results and there are no significant differences of post-haemodialysis BUN and calculated Kt/V between low blood flow technique and bypassing dialysate technique. CONCLUSION: The bypassing dialysate technique is another simpler and practical technique for the routine estimation of post-haemodialysis BUN. PMID- 9351077 TI - A comparison of solute clearance and ultrafiltration volume in peritoneal dialysis with total or fractional (50%) intraperitoneal volume exchange with the same dialysate flow rate. AB - BACKGROUND: The most efficient way to perform automated peritoneal dialysis (APD) has not yet been defined. Tidal peritoneal dialysis (TPD) has been claimed to be more efficient than traditional intermittent peritoneal dialysis (IPD), but few comparative studies have been done keeping dialysate flow the same in the two treatment techniques. METHODS: Six patients were treated with 10, 14 and 24 litres total dialysis fluid volume during 9 h (flow rate 18.5, 25.9 and 44.4 ml/min), receiving the treatments both as IPD and TPD. Glucose concentration in the fluid was held constant during all treatments. Transperitoneal clearances (ml/min) for urea, creatinine and uric acid and ultrafiltration volume was calculated, and comparisons made between TPD and IPD. The total intraperitoneal dwell time was calculated for each treatment session. A peritoneal equilibration test was also done for each patient. RESULTS: The ratio of the creatinine concentration in dialysate to the concentration in plasma at 4 h obtained with the peritoneal equilibration test (PET) averaged 0.77 (range 0.69-0.82). Urea clearance was higher for IPD than for TPD with 10 litres: 14.3 +/- 2.4 and 13.3 +/- 2.7 (P = 0.0092). For 14 and 24 litres urea clearance for IPD and TPD was 16.9 +/- 2.3 and 15.9 +/- 3.5 (n.s.) and 20.9 +/- 3.6 and 19.9 +/- 5.6 (n.s.) respectively. Creatinine clearance was higher for IPD than for TPD with 10 litres: 9.6 +/- 1.3 and 8.9 +/- 1.3 (P = 0.0002). For 14 and 24 litres creatinine clearance for IPD and TPD was 11.0 +/- 0.7 and 9.9 +/- 2.0 (n.s.) and 12.3 +/- 1.2 and 12.4 +/- 2.2 (n.s.) respectively. Uric acid clearance was higher for IPD than for TPD with 10 litres: 8.4 +/- 1.3 and 7.7 +/- 1.0 (P = 0.0054). For 14 and 24 litres uric acid clearance for IPD and TPD was 9.3 +/- 1.7 and 8.9 +/- 2.2 (n.s.) and 11.3 +/- 2.9 and 10.6 +/- 2.6 (n.s.) respectively. IPD gave significantly higher ultrafiltration volume (ml) than IPD for both 10 and 14 litres: 944 +/- 278 and 783 +/- 200 (P = 0.0313) and 1147 +/- 202 and 937 +/- 211 (P = 0.0478). For 24 litres there was no significant difference between IPD and TPD: 1220 +/- 224 and 1253 +/- 256. CONCLUSION: With the lowest dialysate flow rate (18.5 ml/min), solute clearance and ultrafiltration volume was higher on IPD than on TPD. With the intermediate flow rate (25.9 ml/min) the ultrafiltration volume was higher on IPD, but no difference was found for solute clearance. With the highest flow rate (44.4 ml/min) there was no difference neither for ultrafiltration nor for solute clearances. PMID- 9351079 TI - Therapy of post-renal transplantation hyperlipidaemia: comparative study with simvastatin and fish oil. AB - BACKGROUND: Recipients of renal transplantation (RT) exhibit disturbances of serum lipids and apoproteins that may contribute to their cardiovascular morbidity and mortality. In our renal transplant department the hypercholesterolaemia prevalence at the first and fifth year of RT is 70.0% and 81.2%, respectively. Lipid-lowering therapy has been utilized in many Transplant Units. The aim of our study was to evaluate post-RT hyperlipidaemia control with simvastatin or fish oil. METHODS: Forty-three RT patients (26 men and 17 women) with persistent hypercholesterolaemia and stable graft function which were resistant to a lipid-lowering diet (American Heart Association Step Two) were randomized into two groups and treated for 3 months with simvastatin (S) (10mg/day; n = 25) and fish oil (F) (6 g/day; n = 18). Total cholesterol (TC), LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C), lipoprotein a (Lp(a)), apolipoprotein A1 (Apo A1), and apolipoprotein B (Apo B) were monitored and at the study baseline they were similar between the two groups. RESULTS: No side effects were detected after 3 months of therapy. In group S, the concentrations of TC (271 +/- 46 mg% vs 228 +/- 49 mg%; P < 0.001), TG (180 +/- 78 vs 134 +/- 45; P < 0.01), LDL-C (177 +/- 40 vs 144 +/- 43; P < 0.01) and Apo B (96 +/- 18 vs 82 +/- 16; P < 0.001) were significantly reduced, and Apo A1 concentration had increased (135 +/- 24 vs 149 +/- 30; P < 0.01). In group F, the concentrations of TC (266 +/- 25 vs 240 +/- 31; P < 0.001), TG (203 +/- 105 vs 156 +/- 72; P = 0.02) and HDL-C (63 +/- 15 vs 53 +/- 12; P < 0.01) were significantly reduced. CONCLUSIONS: We concluded that low-dose simvastatin and fish oil are both effective and safe in correcting post-RT hyperlipidaemia. Further prospective studies with larger follow-up are needed to clarify whether this therapy has an impact on cardiovascular morbidity and mortality in RT patients. PMID- 9351078 TI - Serum anti-rabbit and anti-horse IgG, IgA, and IgM in kidney transplant recipients. AB - BACKGROUND: The therapeutic efficacy of horse antilymphocyte globulins (ALG) or of rabbit antithymocyte globulins (ATG), used for both the prevention and treatment of allograft rejection has been well documented. However, clinical use of these heterologous antibodies can result in the production of antibodies against horse or rabbit proteins and in the development of serum sickness via circulating immune complexes. METHODS: We studied the production of human IgG, and IgM anti-rabbit and anti-horse globulins, in 240 serum samples from 111 kidney transplant recipients, of whom 89 were treated with ALG or ATG (Merieux France) as prophylaxis. RESULTS: Up to 8.9% of the patients had anti-ALG and/or ATG antibodies before the first transplantation. This proportion increased significantly after. Preimmunization did not appear to be predictive of the occurrence of clinical serum sickness, yet sensitization increased, after transplantation, in up to 71% of the subjects who developed this disorder (P = 0.02). In patients receiving a second transplant, pretransplantation antibody levels were not modified by the immunosuppressive therapy applied. No relationship was found between early rejection and antiglobulin antibodies. CONCLUSIONS: Serum anti-rabbit and/or -horse antibodies were demonstrated in a significant proportion of kidney recipients, even before transplantation, possibly due to environmental exposure. A classical pattern of IgM increase was observed when the patients developed an immune response to ALG or ATG, and an IgA response after ALG. These results suggest that patients receiving ALG/ATG should be monitored for the production of anti-ALG/ATG immunoglobulins. PMID- 9351080 TI - High prevalence of adynamic bone disease diagnosed by biochemical markers in a wide sample of the European CAPD population. AB - BACKGROUND: Adynamic bone disease (ABD) has been described in the current dialysis population to have an unexpectedly high prevalence. Moreover, it is clearly more prevalent in CAPD patients, compared to haemodialysis patients. Recently we demonstrated that both a low (< or = 27 U/1) level of bone alkaline phosphatase (BAP) as determined by an optimized agarose gel electrophoretic technique and a low (< or = 150 pg/ml ) level of iPTH are good markers of ABD with sensitivities of 78.1% and 80.6% and specificities of 86.4% and 76.2% respectively. METHODS: In this study (n = 212), the prevalence of ABD in the European CAPD population was evaluated by means of these biochemical markers. Clinical data on the patients included were recorded at the moment of blood sampling. In patients under CAPD treatment for longer than 9 months, we calculated an index of calcium exposure through PD fluid. RESULTS: In this population with a low exposure to aluminium, the prevalence of ABD as indicated by either a low level of BAP or PTH was 43%. The following risk factors could be identified: advanced age, shorter time on renal replacement therapy, male gender, and high calcium content of PD fluid. The index of calcium exposure was significantly higher in the patients with low BAP and low iPTH levels compared to those with either BAP > or = 27 U/1 or iPTH > 150 pg/ml. The latter finding gives further support to the hypothesis that a high calcium load administered to renal failure patients may lead to 'oversuppressed' parathyroids in ABD. In a subgroup of patients with a high level of BAP associated with a low iPTH level a profile previously shown to be associated in the presence of aluminium overload, significantly higher serum aluminium levels were noted. suggesting that even in patients with low exposure to aluminium, this element still can affect bone metabolism. CONCLUSION: A high prevalence of ABD--as diagnosed by biochemical markers--was observed in the European CAPD population. A number of risk factors could be put forward. The aetiology and pathogenesis of this type of renal osteodystrophy remain to be elucidated, but appear, however, to be multifactorial. PMID- 9351081 TI - Acoustic properties of dialysed kidney by scanning acoustic microscopy. AB - BACKGROUND: A correlation between acquired renal cysts in the dialysed kidney and renal cancer has long been debated, but no changes in the physical properties of kidneys at the microscopic level have been reported. The purpose of the present study was to classify the physical properties of the kidneys of patients undergoing haemodialysis at several stages of pathology by use of the scanning acoustic microscope. METHODS: Sixteen surgically excised kidneys of dialysis patients were investigated. Tissues were fixed in 10% formalin, frozen in acetone, and cut 10 microns thick on a cryostat. We used a scanning acoustic microscope operated in the frequency range of 100-200 MHz. Attenuation constant and sound speed were measured on a two-dimensional distribution. RESULTS: The attenuation constant for inflammatory granulation tissue was significantly higher than that for hyaline degeneration tissue (P < 0.001). Sound speed was high for granulation tissue, but tended to diminish gradually for hyaline degeneration. Sound speed increased again with progression to cystic degeneration (P < 0.001), but the attenuation constant remained low. When a cystic kidney contained a malignant lesion, the previously low attenuation constant rose at that site (P < 0.001), and the previously high sound speed was diminished (P < 0.001). CONCLUSION: Our data suggest that the physical properties of dialysed kidneys at different stages of pathology can be classified by their acoustic properties. Simultaneous evaluation of attenuation constant and sound speed is considered applicable to determining whether tissues contain malignant elements. PMID- 9351082 TI - Delayed onset of membranoproliferative glomerulonephritis in a patient with type I cryoglobulinaemia. PMID- 9351083 TI - Minimal-change disease in association with sarcoidosis. PMID- 9351084 TI - Consider sarcoidosis in patients with nephrocalcinosis, even if the chest roentgenogram is normal. PMID- 9351086 TI - Bilateral retrobulbar optic neuritis with hepatitis B vaccination. PMID- 9351085 TI - Potential beneficial effect of tamoxifen in retroperitoneal fibrosis. PMID- 9351087 TI - Aortoenteric fistula: an unusual cause of gastrointestinal bleeding in a haemodialysis patient. PMID- 9351088 TI - Xanthomonas maltophilia--a growing problem in the haemodialysis population. PMID- 9351089 TI - Salmonella pericarditis and pericardial effusion in a patient with systemic lupus erythematosus on haemodialysis. PMID- 9351090 TI - Opaque enema CT scan allows early diagnosis of non-occlusive right colonic ischaemia in dialysis patients. PMID- 9351091 TI - Treatment of an arteriovenous fistula by the placement of a Z-stent and embolization in a patient with nephrotic syndrome. PMID- 9351092 TI - A renal transplant recipient with acute paraparesis due to an Aspergillus epidural abscess. PMID- 9351093 TI - The dialysed lady with one swollen cheek. PMID- 9351094 TI - The description of the renal glomeruli by Marcello Malpighi. PMID- 9351095 TI - Take the shadow for the substance. PMID- 9351096 TI - Recurrence of autonomous hyperparathyroidism in dialysis patients. PMID- 9351097 TI - Colchicine could be safely used in renal patients. PMID- 9351098 TI - Antiproteinuric effect of losartan in patients with chronic renal diseases. PMID- 9351099 TI - Long-term treatment of IgA nephropathy with cyclosporin A--a preliminary report. PMID- 9351100 TI - Antibody level after hepatitis B vaccination. PMID- 9351101 TI - Uptake of influenza vaccination by patients receiving renal replacement therapy. PMID- 9351102 TI - A survey of hepatitis C virus infection in haemodialysis patients over a 7-year follow-up. PMID- 9351103 TI - MRI of paramedian thalamic stroke with sleep disturbance. AB - The paramedian thalamus is believed to play an important role in the regulation of sleep, and disturbances of sleep regulation are known to occur in paramedian thalamic stroke (PTS). We examined 12 consecutive patients with PTS and sleep disturbance by MRI. Two distinct groups of patients could be defined: six presenting with severe hypersomnia (group 1) and six with slight sleepiness (group 2). On MRI, all patients had ischaemic lesions involving the paramedian thalamic nuclei, the centre of the lesions being the dorsomedial and centromedial thalamic nuclei. In group 1 the lesions were bilateral, butterfly-shaped infarcts involving the paramedian nuclei (three cases); or unilateral with an extension into the subthalamic nuclei. In group 2 the lesions were unilateral and limited to the paramedian nuclei, mainly the dorsomedial nucleus. Bilateral lesions can be attributed to a common origin in some cases for both paramedian thalamic arteries and the mesencephalic arteries. PMID- 9351104 TI - MRI of the brain in patients with miliary pulmonary tuberculosis without symptoms or signs of central nervous system involvement. AB - MRI was performed on patients with miliary pulmonary tuberculosis to look for brain involvement and to study the features sequentially, during treatment. We studied seven patients with typical radiographic tuberculosis, and no symptoms or signs of central nervous system involvement. Conventional spin-echo (SE) imaging, including contrast enhanced images, was performed in all cases. All patients showed brain involvement: four patients showed lesions mainly less than 3 mm in diameter, better seen on contrast-enhanced images. These patients showed oedema around the lesions after 2 months of treatment, with subsequent regression on follow-up. The remaining three patients had multiple lesions, 3 mm or more in diameter, which showed a gradual decrease on follow-up. We conclude that the brain may commonly be involved in miliary pulmonary tuberculosis. The response to treatment depends on the stage of the granuloma and shows a definite pattern of healing on follow-up. PMID- 9351105 TI - Dilatation of deep medullary veins in cortical venous occlusion due to focal tuberculous leptomeningitis. AB - We report a case of surgically proven, focal left parietal tuberculous leptomeningitis. Occlusion of superficial cerebral veins in the affected area led to dilatation of medullary veins to drain the left parietal lobe. Deep medullary veins were clearly demonstrated on MRI and angiography. PMID- 9351106 TI - Human African trypanosomiasis: MRI. AB - We report a case of human African trypanosomiasis caused by Trypanosoma brucei rhodesiense. After the febrile period of parasite dissemination, the patient had meningeal involvement but normal CT. MRI showed the appearances of meningitis. After two periods of arsenical treatment, a severe encephalopathy occurred suggesting post-therapeutic reactive encephalitis (PTRE). Nevertheless, T2 weighted MRI showed no oedema, but focal bilateral high signal areas in the white matter. PTRE was excluded and a third course of treatment was undertaken. The lesions progressively disappeared. PMID- 9351107 TI - Cyclosporine-related reversible posterior leukoencephalopathy: MRI. AB - Three patients aged 48, 11 and 40 years, two of whom were recent recipients of renal transplants and one of a bone marrow transplant, developed seizures, with cortical blindness in two cases. All were immunosuppressed with cyclosporine and were hypertensive at the onset of symptoms. MRI showed predominantly posterior signal changes in all three cases. The abnormalities were more conspicuous on fast FLAIR images than on conventional T2-weighted spin-echo images. PMID- 9351108 TI - Primary germinoma of the posterior cranial fossa: a case report. PMID- 9351109 TI - Imaging and pathological features of primary malignant rhabdoid tumours of the brain and spine. AB - In this article two cases of primary malignant extrarenal rhabdoid tumour are described. In the affected children the brain and the spinal cord were the primary sites of origin of the tumour. The imaging findings are presented and the pathology discussed. Although the imaging features are non-specific, rhabdoid tumour should be included in the differential diagnosis of childhood intracranial and spinal neoplasms. PMID- 9351110 TI - MRI of autosomal dominant pure spastic paraplegia. AB - We examined 16 patients with autosomal dominant pure spastic paraplegia (HSP) and 15 normal controls matched for age and sex using MRI of the brain and spinal cord. Images were assessed qualitatively by two independent radiologists, blinded to the clinical diagnosis. Areas of the brain and corpus callosum on one midsagittal slice and the area of the brain on one axial slice were measured and a "corpus-callosum index" expressing the size of the corpus callosum relative to that of the brain was calculated. Cross-sectional areas and anteroposterior and transverse diameters of the spinal cord at the levels of C 2, C 5, T 3, T 6, T 9 and T 11 were measured. No significant differences between patients and controls were found on qualitative evaluation of the images. The patients had a significantly smaller corpus callosum and "corpus-callosum index" than controls. This finding, not reported previously, might indicate that the disease process in pure HSP is not confined to the spinal cord. The anteroposterior diameters of the spinal cord at T 3 and T 9 were significantly smaller in patients than in controls. This might correspond to the degeneration of the pyramidal tracts and the dorsal columns described at neuropathological examination. PMID- 9351111 TI - Suitability of cerebrospinal fluid as a signal-intensity reference on MRI: evaluation of signal-intensity variations in the lumbosacral dural sac. AB - The suitability of the cerebrospinal fluid (CSF) in the lumbosacral dural sac as an internal signal-intensity reference was studied on magnetic resonance imaging (MRI) of the lumbar spine using a surface coil and motion artefact suppression technique. A signal-intensity reference is needed when signal is compared between images, studies or subjects. Homogeneity of the CSF was estimated visually on T2 weighted images of 60 subjects at 1.5 T and of another 60 subjects at 0.1 T. Spines with a severely narrowed dural sac or marked scoliosis were excluded from the study to avoid partial volume effect. CSF was homogeneous in 82% and 73% of the examinations at 1.5 T and 0.1 T, respectively. The type and location of the local inhomogeneities did not relate to local narrowings of the dural sac. The signal intensity of CSF was measured in 108 examinations at 0.1 T after correcting the spatially-dependent signal-intensity non-uniformities with a phantom-based method. The signal-intensity difference between the CSF in the upper and lower lumbar dural sac was less than 10% in 73% of the examinations. The CSF in the lumbosacral dural sac can be a useful signal-intensity reference for estimation of the signal of the adjacent structures in patients without severe narrowing of the dural sac or marked scoliosis. It may contribute to assessing spinal disease processes. PMID- 9351112 TI - Imaging of spinal cord compression due to thoracic extramedullary haematopoiesis in myelofibrosis. AB - We describe a case of spinal cord compression secondary to extramedullary haematopoiesis in a patient with primary myelofibrosis. We show that MRI should be the procedure of choice for patients suspected of this condition. Furthermore, it could be of value for assessing the extent of cord compression, planning radiotherapy and for follow-up. PMID- 9351113 TI - Spontaneous spinal epidural haematomas with repeated remission and relapse. AB - We present a case of spontaneous spinal epidural haematoma (SSEH) with a rare clinical course of repeated spontaneous recovery and relapse. The patient suffered three episodes of upper-back pain of sudden onset followed by sensory and motor dysfunction after weight lifting. In the first two episodes, the neurological deficits recovered spontaneously and completely. In the last episode, paraplegia persisted even after emergency surgery. Serial studies with computed tomographic (CT) myelography and magnetic resonance imaging (MRI) demonstrated the remitting and relapsing course of the SSEHs. The possible causes of the SSEHs and the mechanisms of spontaneous recovery are discussed. PMID- 9351114 TI - Nasopharyngeal carcinoma: MRI and CT assessment. AB - Precise assessment of the extent of nasopharyngeal carcinoma (NPC) represents the basic step towards optimal treatment. We compared the capacity of CT and MRI in assessing the extent of NPC in 67 patients. MRI was superior to CT in demonstrating lesions in the retropharyngeal node, skull base, intracranial area, carotid space, longus colli muscle and levator palatini muscle. Of 25 cases in which retropharyngeal adenopathy was recognised only on MRI, seven had been reported as showing oropharyngeal involvement and 18 as primary extension to the carotid space on CT. MRI showed skull-base involvement in 40 patients compared with 27 on CT and intracranial involvement in 38 patients versus 24 on CT. There was not a single case in which skull base invasion was seen on CT but not on MRI. MRI enabled improved recognition of tumour infiltration of longus colli muscles (34 cases compared with 15 on CT). It allowed us to clarify 12 questionable sinonasal opacities on CT. Overall, T-staging was changed in 18 of 67 patients (26.9%), including upstaging in 15 cases and down-staging in 3 cases, after comparing CT with MRI. The nodel status was changed from negative on CT to positive on MRI in 4 of 67 patients (6%). We believe that MRI allows more accurate evaluation of the extent of NPC than CT and should be the primary mode of investigation. PMID- 9351115 TI - Neurosarcoma of the face: MRI. AB - Neurosarcoma is a rare tumour originating from the sheath of peripheral nerves. Facial lesions have been reported in about 20 patients. We describe the MRI appearances of neurosarcoma with histological correlation in three patients. The lesions lay in the submandibular region, the left parapharyngeal space and the right orbit. MRI showed a well-defined mass with mixed components. The lesions were moderately heterogeneous on T1-weighted images in two cases and on T2 weighted images in all cases. Gadolinium enhancement occurred in all cases to variable degrees. In two cases, small high signal foci were seen on T2-weighted sequences. MRI appearances of neurosarcoma are not specific. PMID- 9351116 TI - Repeat intra-arterial papaverine for recurrent cerebral vasospasm after subarachnoid haemorrhage. AB - We assessed the prevalence of recurrent vasospasm following failure of intra arterial papaverine and the efficacy of repeat intra-arterial infusions of papaverine for control of recurrent vasospasm. Of 24 patients treated with intra arterial papaverine for vasospasm following aneurysm surgery, 12 did not improve clinically after the initial treatment; 9 received second or third infusions on consecutive days; 6 received only a second infusion; and 3 received a third. Superselective infusion into the intracranial arteries was performed in all nine cases. Despite angiographic improvement after the initial or second infusions, all nine patients showed varying degrees of recurrent vasospasm at the time of the second or third treatment. Within 24 h of a second infusion, three of the six patients had significant clinical improvement, and one of these showed marked improvement soon after a third infusion. Our preliminary results suggest that repeat papaverine infusion may be a way of controlling recurrent or recalcitrant vasospasm. PMID- 9351117 TI - Screening for colorectal cancer. PMID- 9351118 TI - Circannual rhythm of measles subsequent to immunisation. AB - This is an epidemiological study of biennial and annual rhythms of measles in England and Wales between 1959-1994 highlighting changes following immunisation. The study describes annual rhythms starting in autumn (Week 41) and the statistical method tests the fit of a sine curve to annual data. Before immunisation the previously established biennial rhythm was seen: the two years differed in magnitude of seasonal variation, position of the peak, "visibility of school holidays" and peak breadth. After immunisation (1968) these biennial features tended to disappear, one annual rhythm becoming the predominant feature. By the late eighties adjacent years had very similar sine curve fits. About 1990 (89-91) the fit of the sine curve was no longer significant, was of low amplitude and numbers were at an all time low. In 1992 significant annual variation returned, numbers rose and magnitude of seasonal variation increased. At this time a new epidemic was being predicted on other evidence and this renewed sine curve fit may be an additional warning signal. A possible influence of normal birth rhythm on numbers of susceptibles is described. PMID- 9351119 TI - The management of first seizures in adults in a district general hospital. AB - This study considered the management of first seizures in adults in Stirling Royal Infirmary over a six month period. Thirty-four patients presented of whom 19 were admitted to medical wards. Alcohol was implicated in 35% of cases. Blood tests were done in many but provided little useful information. CT Scan was performed in 53% and was abnormal in 15% (five patients). EEG was requested for 21% and failed to influence management in any. Six patients (18%) were started on anticonvulsant therapy. It was recorded in only three cases that advice on driving had been given. The literature concerning single seizures is complex, especially with regards to recurrence risk and treatment benefits. We await with interest the publication of the SIGN (Scottish Intercollegiate Guidelines Network) guidelines for seizure investigation and treatment in Scotland. PMID- 9351120 TI - The neonatal consequences and neonatal cost of reducing the number of embryos transferred following IVF. AB - This clinical audit project examined the effects of change of policy between 1990 and 1993 transferring an average two (maximum three for particular cases) embryos to women undergoing IVF in the West of Scotland programme. All women who achieved clinical pregnancy in 1990 (92 women) and 1993 (93 women) as a result of the IVF programme were included in the study. The hospital records of women via the programme were analysed. The results of the study showed that there was a significant reduction in the rate of multiple pregnancy, preterm birth and low birth weight babies in the 1993 group (new policy). The cost of neonatal intensive care in 1993 for babies born following IVF was about nine times lower than that in 1990 (old policy). This study concluded that a policy of transferring two embryos (or three for particular cases) to women in an IVF programme, had improved the perinatal outcome and reduced the cost of the neonatal service for those babies. PMID- 9351121 TI - Treatment of severe acute vascular rejection in a renal allograft with mycophenolate mofetil and high dose steroids. AB - A case of a highly sensitised haemodialysis patient who developed severe vascular rejection in her third renal allograft is presented. This severe rejection episode responded to mycophenolate mofetil (MMF) and high dose steroids. PMID- 9351122 TI - Audit of acute upper gastrointestinal haemorrhage: the effects of education and the introduction of a protocol. AB - The aim of this audit was to monitor the effects of the introduction of a protocol for the management of acute upper GI haemorrhage and a teaching programme for House Officers in Ninewells Hospital Dundee. All patients admitted to hospital with a history of acute haematemesis or melaena were included in the study and purpose designed audit forms were completed on all patients. Following an initial six month audit period the protocol and teaching were introduced. A further six months audit was then performed. A total of 310 patients were audited over the two six month periods. The results suggest that the introduction of a management protocol and training, in conjunction with a policy of active endoscopic intervention may reduce the number of out of hours emergency endoscopies and the need for emergency surgery. PMID- 9351123 TI - Glasgow resurrectionists. AB - The Napoleonic Wars and the colonial campaigns of the early 1800s created a great need for surgical training. Many of the cadavers used in Glasgow's schools of Anatomy were resurrected from local churchyards or imported from Ireland. In the 1820s, the activities of some resurrectionists showed gross insensitivity, with bodies being stolen before the funeral. In the early 1830s, cholera riots and the fear of "burking" led to the Anatomy Bill of 1832 receiving the Royal Assent. PMID- 9351124 TI - Therapeutic potential of S-nitrosothiols as nitric oxide donor drugs. PMID- 9351125 TI - Recreational overdosage of carbamazepine in Paisley drug abusers. PMID- 9351126 TI - Pontine microinjection of carbachol does not reliably enhance paradoxical sleep in rats. AB - It has been repeatedly shown in cats that acute administration of carbachol into the pontine reticular formation (PRF) readily evokes a state that closely mimics natural paradoxical sleep (PS). Surprisingly, there are few corresponding studies in rats. In order to further characterize the effects of pontine carbachol in rats, 151 injections of different doses (from 3 micrograms to 0.005 microgram in 0.1 microliter saline) of carbachol were made at different sites within the PRF of 70 rats. Sleep-waking states obtained in the 4 hours following carbachol administration were compared to control values, obtained both under baseline condition (no injection) and following pontine injection of 0.1 microliter saline. On the one hand, from the whole set of carbachol injections, it appeared that: 1) most injections (112/151) did not significantly alter the sleep-wake states; 2) when carbachol was effective, it induced either increased PS (20 injections) or increased waking (19 injections); and 3) effective injection sites were intermingled with noneffective sites. Dose- or site-dependency effects can account in part, but not totally, for these discordant results. On the other hand, in accordance with previous rat studies, we found that: 1) the PRF medial and ventral to the motor trigeminal nucleus was the most effective region for carbachol to increase PS; 2) carbachol-induced PS enhancement was of moderate magnitude (+60% above control saline level over the 4-hour recording time); 3) latency to onset of the first PS episode was not shortened; and 4) only the number of PS episodes was increased, their duration was not prolonged. These characteristics of carbachol-induced PS enhancement strongly differ, both in terms of magnitude and timing, from those described in cats. We suggest that the less reliable and weaker effects of pontine carbachol injection in rats compared to cats can be due to methodological problems inherent in the intracerebral microinjection technique and also to species-related differences in the mechanisms controlling the PS state. PMID- 9351127 TI - Sleep-disordered breathing and motor vehicle accidents in a population-based sample of employed adults. AB - Studies have consistently shown that sleep apnea patients have high accident rates, but the generalizability of the association beyond clinic populations has been questioned. The goal of this investigation was to determine if unrecognized sleep-disordered breathing in the general population, ranging from mild to severe, is associated with motor vehicle accidents. The sample comprised 913 employed adults enrolled in an ongoing study of the natural history of sleep disordered breathing. Sleep-disordered breathing status was determined by overnight in-laboratory polysomnography and motor vehicle accident (MVA) history was obtained from a statewide data base of all traffic violations and accidents from 1988 to 1993. Men with five or more apneas and hypopneas per hour of sleep [apnea-plus-hypopnea index (AHI) > 5], compared to those without sleep-disordered breathing, were significantly more likely to have at least one accident in 5 years (adjusted odds ratio = 3.4 for habitual snorers, 4.2 for AHI 5-15, and 3.4 for AHI > 15). Men and women combined with AHI > 15 (vs. no sleep-disordered breathing) were significantly more likely to have multiple accidents in 5 years (odds ratio = 7.3). These results, free of clinic selection bias, indicate that unrecognized sleep-disordered breathing in the general population is linked to motor vehicle accident occurrence. If the association is causal, unrecognized sleep-disordered breathing may account for a significant proportion of motor vehicle accidents. PMID- 9351128 TI - Sleep in a truck berth. AB - The aim of the present pilot study was to study the effects of sleep in a truck berth. Experiment A included eight subjects who slept during two conditions (laboratory and in a truck berth during quiet conditions). Experiment B included two conditions (truck-berth sleep during quiet and noisy/disturbed conditions, respectively); six subjects participated. Polysomnography was recorded and ratings of sleep quality and postsleep sleepiness were made. During the truck berth conditions, noise was continuously recorded. When two-tailed t tests were used, the results showed no significant effects (alpha level = 0.05) for any of the experiments. However, when one-tailed tests were used, experiment A showed a longer rapid eye movement (REM) latency for the truck-berth condition. Experiment B showed less-refreshing sleep for the disturbed condition (one-tailed test). The noise level was significantly higher during the disturbed condition. The results showed that electroencephalograph (EEG)-recorded sleep was not affected by sleeping in a truck, even when the truck was parked at a noisy location (truck terminal). However, considering some limitations of the experiments, for example small sample size, lack of adaptation night, etc., the present results should be interpreted with some caution and need to be replicated. PMID- 9351129 TI - Value of the multiple sleep latency test (MSLT) for the diagnosis of narcolepsy. AB - Since its introduction, the multiple sleep latency test (MSLT) has played a major role in the diagnosis of narcolepsy. We assessed its diagnostic value in a series of 2,083 subjects of whom 170 (8.2%) were diagnosed with narcolepsy. The sensitivity of the combination of two or more sleep onset rapid eye movement (REM) periods (SOREMPs) with a mean sleep latency of < 5 minutes on an initial MSLT was 70% with a specificity of 97%, but 30% of all subjects with this combination of findings did not have narcolepsy. In some narcoleptics who had more than one MSLT, the proportion of naps with SOREMPs varied substantially from the initial MSLT to the follow-up test. The highest specificity (99.2%) and positive predictive value (PPV) (87%) for MSLT findings was obtained with the criteria of three or more SOREMPs combined with a mean sleep latency of < 5 minutes, but the sensitivity of this combination was only 46%. The combination of a SOREMP with a sleep latency < 10 minutes on polysomnography yielded a specificity (98.9%) and PPV (73%) almost equal to those obtained from combinations of MSLT findings, but the sensitivity was much lower. Our results suggest that the MSLT cannot be used in isolation to confirm or exclude narcolepsy, is indicated only in selected patients with excessive daytime sleepiness, and is most valuable when interpreted in conjunction with clinical findings. PMID- 9351130 TI - MRI findings in narcolepsy. AB - The neuropathology of narcolepsy is unknown. Recently, Plazzi et al. (1) reported magnetic resonance imaging (MRI) abnormalities in the pontine tegmentum of three patients with long-standing idiopathic narcolepsy. Considering the localization of the neuroradiological findings in the pontine reticular formation, where rapid eye movement (REM) sleep is generated, the authors suggested a causal relationship between narcolepsy and MRI abnormalities. Frey and Heiserman, however, found pontine MRI abnormalities in only two of 12 patients with narcolepsy both of whom had long-standing hypertension (2). Pullicino et al. noted similar pontine MRI abnormalities in patients with subcortical arteriosclerotic encephalopathy-like ischemic rarefaction of the pons (3). Thus, the changes noted by Plazzi et al. may have been caused by small-vessel disease rather than narcolepsy. To assess whether altered pontine MRI signals are a regular feature of idiopathic narcolepsy, we selected randomly from our database seven patients with narcolepsy with cataplexy. Of these seven, three agreed to have brain MRIs; their cases are described below. None had pontine MRI abnormalities. PMID- 9351131 TI - The effect of cutaneous and deep pain on the electroencephalogram during sleep- an experimental study. AB - The interaction between sleep and pain has been insufficiently studied, and no experiments have investigated whether pathologic sleep patterns as seen in pain patients can be replicated experimentally by well-defined pain stimuli. An experimental model would therefore be valuable for further studies on the interaction between pain and sleep. In this study, three well-defined experimental stimuli (muscle, joint, and cutaneous pain) were applied during sleep, and the electroencephalogram (EEG) pattern was quantified. The pain stimuli were applied during slow-wave sleep in 10 healthy subjects. Using nine surface recordings, the EEG was sampled before and during pain stimuli. Frequency analysis was performed, resulting in 10 EEG features describing the responses to pain. During the muscle-pain stimulus an arousal effect was observed and a decrease in delta (0.5-3.5 Hz) and sigma (12-14 Hz) as well as increases in alpha 1 (8-10 Hz) and beta (14.5-25 Hz) activities were seen. During joint pain, however, more universal EEG changes were seen with a decrease in the lowest frequency bands [delta, theta (3.5-8 Hz) and alpha 1] and an increase in the higher frequencies [alpha 2 (10-12 Hz), sigma and beta bands]. No background EEG changes were observed during the cutaneous stimulus. There were several differences in the responses from the nine EEG channels, but no derivation seemed especially sensitive to detect the evoked changes. The study highlights the complexity of pain on the sleep EEG. The experimental model has shown that pain from different body structures, as well as signals from various EEG derivations, may give different responses in sleep microstructure. PMID- 9351132 TI - Sleep scoring at a lower resolution. AB - Sleep scoring of whole-night polysomnograms is labor intensive. Scoring fewer epochs saves labor at the cost of accuracy; this study investigates the trade-off between the two. Whole-night sleep measures of 12 patients with sleep apnea syndrome, 10 patients with narcolepsy, and 35 controls were first computed using conventional successive 30-second epochs. Using the resulting list of sleep stages, a variable number of epochs was skipped among remaining epochs; the measures were recomputed for the reduced lists. The Bland-Altman analysis was used to define the agreements among the sleep measures at the conventional resolution and those at the lower resolutions. Scoring one-half to one-third of the number of epochs changes the duration of sleep stages only up to 2.5% and 5%, respectively, for all groups and sleep stages. In apnea patients, rapid eye movement (REM) latency deviates < 15 minutes when half of the epochs are scored. In controls and narcoleptics, much lower resolutions can be used before reaching the same level. Potential restrictions for the application of the method are discussed. PMID- 9351133 TI - Reduced hospitalization with cardiovascular and pulmonary disease in obstructive sleep apnea patients on nasal CPAP treatment. AB - Cardiovascular and pulmonary disease (CVPD) is common in patients with obstructive sleep apnea syndrome (OSAS). This retrospective study addressed the accumulated in-hospital time during 2 years prior to treatment with nasal continuous positive airway pressure (nCPAP) as compared to 2 years after initiating of nCPAP in patients with OSAS and CVPD. A cohort representing all patients (n = 88) receiving nCPAP during the period 1988-1994 at the Skovde Central Hospital, Skovde, Sweden, was studied. Data collection was based on interviews with patients as well as reviews of clinic charts. All hospitalizations and diagnostic codes by any type were thereby successfully gathered for the whole group. Six patients with confounding serious diseases were excluded from the analysis. A CVPD diagnosis (ICD-9, codes 401-435 and 490-496) was found in 54 out of 82 patients (66%), of whom 36 of 58 were nCPAP users (62%) and 18 of 24 were nonusers (75%). In 54 sleep apneics with CVPD, 31 were hospitalized acutely under one or more of these diagnostic codes during the study period of 4 years. The total number of in-hospital days due to CVPD in the nCPAP users (n = 19) before nCPAP prescription was 413 days (median 10, range 3-66) compared to 54 days (median 0, range 0-25) after nCPAP (p < 0.0001). The corresponding values for the nonuser group (n = 12) was 137 days (median 8.5, range 0-42) before and 188 days (median 9.5, range 0-47) after the nCPAP prescription (ns). We conclude that nCPAP treatment reduces the need for acute hospital admission due to CVPD in patients with OSAS. This reduction of concomitant health care consumption should be taken into consideration when assessing the cost-benefit evaluation of nCPAP therapy. PMID- 9351134 TI - Respiratory arousal from sleep: mechanisms and significance. AB - The mechanisms by which respiratory stimuli induce arousal from sleep and the clinical significance of these arousals have been explored by numerous studies in the last two decades. Evidence to date suggests that the arousal stimulus in nonrapid eye movement sleep (NREM) is related to the level of inspiratory effort rather than the individual stimuli that contribute to ventilatory drive. A component of the arousal stimulus proportional to the level of inspiratory effort may originate in mechanoreceptors either in the upper airway or respiratory pump. Medullary centers responsible for ventilatory drive may also send a signal proportionate to the level of drive to higher centers in the brain which are responsible for arousal. Thus, the arousal stimulus may consist of multiple components, each increasing as inspiratory effort increases. The level of effort triggering arousal is an index of the arousability of the brain (arousal threshold). A deeper stage of sleep, central nervous system depressants, prior sleep fragmentation, and the presence of obstructive sleep apnea (OSA) have been observed to increase the arousal threshold to airway occlusion. Less information is available concerning the mechanisms of arousal from rapid eye movement (REM) sleep. While REM sleep is associated with the longest obstructive apneas in patients with OSA, normal human subjects appear to have a similar or lower arousal threshold to respiratory stimuli in REM compared to NREM sleep. Recent studies have challenged the assumption that the termination of all obstructive apnea is dependent on arousal from sleep. Improvements in methods to detect and quantitate changes in the cortical electroencephalogram (EEG) may better define the relationship between arousal and apnea termination. This may result in improved criteria for identifying EEG changes of clinical significance. While little is known concerning the mechanisms of arousal in central sleep apnea, arousal may play an important role in inducing this type of apnea in some patients. PMID- 9351136 TI - Bibliography of recent literature in sleep research. PMID- 9351135 TI - Epworth Sleepiness Scale in a sample of the Spanish population. PMID- 9351137 TI - Undercover careseekers: simulated clients in the study of health provider behavior in developing countries. AB - The simulated client method (SCM) has been used for over 20 years to study health care provider behavior in a first-hand way while minimizing observation bias. In developing countries, it has proven useful in the study of physicians, drug retailers, and family planning services. In SCM, research assistants with fictitious case scenarios (or with stable conditions or a genuine interest in the services) visit providers and request their assistance. Providers are not aware that these clients are involved in research. Simulated clients later report on the events of their visit and these data are analyzed. This paper reviews 23 developing country studies of physician, drug retail, and family planning services in order to draw conclusions about (1) the advantages and limitations of the methods; (2) considerations for design and implementation of a simulated client study; (3) validity and reliability; and (4) ethical concerns. Examples are also drawn from industrialized countries, related methodologies, and non health fields to illustrate the issues surrounding SCM. Based on this review, we conclude that the information gathered through the use of simulated clients is unique and valuable for managers, intervention planners and evaluators, social scientist, regulators, and others. Areas that need to be explored in future work with this method include: ways to ensure data validity and reliability; research on additional types of providers and health care needs; and adaptation of the technique for routine use. PMID- 9351138 TI - Images of health and health care options among low income women in Punjab, Pakistan. AB - We studied women's beliefs and experiences of health and health care among 42 women in an urban slum area in Lahore and a village 40 km outside of Lahore. Data were collected through repeated, in-depth interviews in Urdu or Punjabi totalling 200 hours. For triangulation purposes, four focus group discussions with additional women were performed, as well as in-depth interviews with eight mothers-in-law, three traditional practitioners and three medical practitioners. The women's images of health reflected expectations on the women in society. Women from the village and women from the lowest socioeconomic stratum (SES) spoke of health in terms of physical strength; women from the city and women from low SES spoke of health in terms of mental strength; and women from medium SES discussed it in terms of cultural competence. Overall, health had a very low priority in these women's lives. Two health problems were reported by all women: mental tension leading to headache and white vaginal discharge leading to body pains and fatigue. These health problems were seen as part of womanhood; if treatment was sought, it was often from traditional healers. Village women had a flexible, pragmatic attitude toward health care resources and used all types until treated. Their relation to the doctor was specific; they were mostly concerned with the medical treatment. In contrast, city women chose health care providers depending on type of illness, and being met with respect was for them of equal concern. Childbearing experiences influenced the perceptions of health and health care. Mothers of daughters were seen to both need and deserve less food, health care and attention. These mothers were less vocal about health complaints. Women without children spoke of health in terms of physical strength. These women may have less access to health care because children cannot be used as an "excuse", and because they are not worth spending resources on. PMID- 9351139 TI - Developing community mental health services for children in South Africa. AB - As a result of South Africa's Apartheid history, mental health care for black people, especially in rural areas, has been grossly inadequate and even non existent in many areas. Children have been severely neglected in this regard. This paper describes an attempt by clinical psychologists to develop a community intervention programme for children with emotional problems. From their hospital base the authors set out, on a monthly basis, to outlying areas up to 250 km away to (1) train primary care nurses and other personnel in the basic techniques of identifying and dealing with uncomplicated psychological problems of childhood, and (2) render consultations to psychologically disturbed children. The paper argues the need to provide primary care workers with mental health skills and thus integrate childhood mental health care into the primary care structure. Such a move could make mental health care accessible to all inhabitants, thus deviating from the policies of the past. PMID- 9351140 TI - A study of the breast cancer dynamics in North Carolina. AB - This work is concerned with the study of breast cancer incidence in the State of North Carolina. Methodologically, the current analysis illustrates the importance of spatiotemporal random field modelling and introduces a mode of reasoning that is based on a combination of inductive and deductive processes. The composite space/time analysis utilizes the variability characteristics of incidence and the mathematical features of the random field model to fit it to the data. The analysis is significantly general and can efficiently represent non-homogeneous and non-stationary characteristics of breast cancer variation. Incidence predictions are produced using data at the same time period as well as data from other time periods and disease registries. The random field provides a rigorous and systematic method for generating detailed maps, which offer a quantitative description of the incidence variation from place to place and from time to time, together with a measure of the accuracy of the incidence maps. Spatiotemporal mapping accounts for the geographical locations and the time instants of the incidence observations, which is not usually the case with most empirical Bayes methods. It is also more accurate than purely spatial statistics methods, and can offer valuable information about the breast cancer risk and dynamics in North Carolina. Field studies could be initialized in high-rate areas identified by the maps in an effort to uncover environmental or life-style factors that might be responsible for the high risk rates. Also, the incidence maps can help elucidate causal mechanisms, explain disease occurrences at a certain scale, and offer guidance in health management and administration. PMID- 9351141 TI - Collecting retrospective data: accuracy of recall after 50 years judged against historical records. AB - Recent interest in a lifecourse perspective on health inequalities will rekindle concerns about the accuracy of retrospective data. The present paper demonstrates that recalled information on some types of social circumstances can be obtained with a useful degree of accuracy using an interview technique which helps to minimize recall bias. Lifegrid information about social circumstances during their youth and childhood was collected from 57 subjects in early old age and compared with archive material of the same subjects' social circumstances recorded 50 years previously. A comparison of interview with archive data revealed that a substantial majority of subjects had recalled simple socio demographic information after a period of 50 years with a useful degree of accuracy. Within lifecourse research, it is concluded, carefully collected retrospective data offer a valuable complement to birth cohort studies, provided that such usage is sensitive to the types of items of information which can, and can not, be recalled accurately. PMID- 9351142 TI - Ageing and the relationship between functional status and self-rated health in elderly men. AB - Functional status (measured as functional limitations or disabilities) is an important determinant of self-rated health in the elderly. Several issues which are not yet clear in this association are addressed in this study: (i) the modifying effect of age on the association; (ii) the effect of recent changes in disability level on the current level of self-rated health, and (iii) the effect of functional limitations on self-rated health, independent of disabilities. Data were derived from the 1990, 1993 and 1995 surveys of the Zutphen Elderly Study, a longitudinal health study in men born between 1900 and 1920. Analyses of repeated measurements were performed with self-rated health as dependent variable and disabilities, functional limitations, age, survey year, and interaction terms as independent variables. Odds ratios were calculated from these models. Men with disabilities in instrumental activities of daily living had no different health ratings than men without disabilities. Those with disabilities in mobility and basic activities of daily living, however, had an odds ratio on poor self-rated health of 4.7 (95% confidence interval: 2.7-7.9) and 8.9 (4.6-17.1) respectively. This association became weaker with increasing age, leading to an absence of a significant association in the oldest group. The current level of self-rated health was only associated with the current level of disabilities. Information on previous levels of disabilities did not contribute to current self-rated health. Functional limitations had a small, but significant, effect on self-rated health when disabilities were taken into account. This study helps in enhancing insight in the complex relationship between functional status and self-rated health in the elderly. PMID- 9351143 TI - Unintended effects of a cost-containment policy: results of a natural experiment in Germany. AB - In this paper empirical evidence for substitution processes caused by the budget for drugs prescribed by office-based physicians is provided. Due to substitution processes in a natural experiment the number of referrals and hospital admissions increased significantly after the introduction of a drug budget in Germany. This leads to additional direct and indirect cost for the health care system. PMID- 9351144 TI - Decision-making and hormone replacement therapy: a qualitative analysis. AB - Biomedical discourse dominates the research literature and media accounts of menopause. Middle aged women are increasingly faced with decisions about hormone replacement therapy (HRT) in the context of differing constructions of menopause and often inconclusive information. There is an apparent discrepancy between the beneficial claims made for HRT in the medical literature and the numbers of women who use it in the U.K. An educational approach has been advocated which assumes that with adequate information more women will take and adhere to HRT. Middle aged women's own views and opinions about medication, health and menopause have been relatively neglected. The extent to which women use medical discourse in discussions of menopause and the extent to which menopause has become "medicalized" remain unclear. This is a descriptive study using in-depth interviews and a qualitative methodology to investigate women's accounts of their decisions relating to HRT use. Ninety-three 50 year old women were recruited from the age/sex register of a North London general practice; 45 women agreed to take part. Three main themes were identified in the women's accounts of their decisions: (1) the presence or absence of troublesome vasomotor symptoms, (2) doctors' views and advice, and (3) views toward menopause and medication. There was a general preference not to take medication, particularly for menopause, which was seen as a natural process unless severe symptoms were present. Women appeared to be considering different criteria from health professionals when making decisions about HRT. Whether these accounts are voiced in a medical consultation will partly depend upon the doctor's beliefs and communication skills as well as the assertiveness of the woman herself. PMID- 9351145 TI - Health-related behaviours and psycho-social characteristics of 18 year-old Australians. AB - Psychosocial variables associated with health-related behaviours for diet, physical activity, alcohol consumption and smoking were examined in 18 year-old Australian men (n = 301) and women (n = 282). These psychosocial variables included Type A behaviour and depression, perceived self-efficacy for engaging in healthy behaviours and perceived barriers to performing these behaviours. Self efficacy for following a healthy diet and moderating alcohol intake was greater in females but males had higher self-efficacy for physical activity. Self efficacy for smoking did not differ according to gender. Lack of willpower was perceived as a barrier to desirable dietary, smoking and physical activity behaviours. Other perceived diet-related barriers included buying suitable foods when eating out, ignorance about appropriate foods and, in young women, perceived expense. Barriers for desirable levels of physical activity included planning time, tiredness, limiting social life and lack of social support. Social occasions were the main perceived barriers preventing both alcohol moderation and quitting smoking. Lack of family support, stress and concerns about weight gain, particularly in women, were perceived barriers to smoking cessation. Type A behaviour was associated with smoking and "unsafe" drinking in both men and women, generally unhealthy dietary choices in young women but with greater physical activity in young men. Depressive affect was significantly higher in female smokers and "unsafe" drinkers and tended to have an inverse relationship with physical activity in men and women. Depressive affect was inversely related to self-efficacy in both men and women for each of the health behaviours examined. Health promotion in young adults should therefore attempt to increase self-efficacy and address perceived barriers to change, taking into account gender-related differences in attitudes and the influence of depression and Type A characteristics on health-related behaviours. PMID- 9351146 TI - Suicide: qualitative data from focus group interviews with youth. AB - Suicide is a leading cause of morbidity and mortality among people aged 15-24 years of age. This paper illustrates the use of focus groups with young people to enhance knowledge of ways to address youth suicide. Analysis of the findings identified three themes perceived by participants as being warning signs of a suicidal friend (personality changes, risk-taking behaviour and unusual actions). An important finding, which has implications for the planning of further suicide prevention strategies, was that young people would either cope alone or turn to a friend if they were feeling suicidal. The fact that a lack of knowledge was identified as the major barrier to youth using existing services/resources suggests that health promotion awareness campaigns which provide information on where young people could access help need to be developed. The use of focus groups with young people has provided valuable insights into ways to address youth suicide. We urge other researchers to incorporate similar methodologies. PMID- 9351147 TI - The rise and rise of proton pump inhibitor drugs: patients' perspectives. AB - The prescribing of proton pump inhibitor (PPIs) drugs has increased by 456% in the past 4 yr, despite no evidence of increased morbidity for gastrointestinal conditions. There has been no full explanation for this dramatic increase. Doctors attribute the rapid and apparently unjustified increase in prescribing of this new and expensive drug to patients' demands and not to the extensive advertising campaign. This paper presents the findings from interviews with 20 patients who were taking PPIs. The results revealed that the prescribing of PPIs was mainly initiated in primary care, with little evidence of overt patient demand for PPIs, influenced by the media or social contacts. Patients' perceptions and beliefs about PPIs are explored and discussed. Patients modified the prescribed regimen to suit their perceived needs. Patients felt PPIs were more effective than other drugs they had tried previously and expressed their concerns about stopping PPIs, or changing to another drug. However, despite these reservations, the majority of patients interviewed said they would change if their general practitioner (GP) suggested it. PPIs led some patients to abandon, or not to attempt, lifestyle changes. The consequences of this and the implications of the continued prescribing of PPIs are discussed. PMID- 9351148 TI - Sex differences in prescribed medications: another case of discrimination in general practice. AB - Biological, social and behavioural factors influence doctors to prescribe different types of medications to male and female patients. Secondary analysis of data from the Australian Morbidity and Treatment Survey 1990-1991 was conducted using multiple logistic regression to discriminate male and female patient encounters in general practice. The approach used considered possible confounding influences of GP and patient characteristics. The results showed that females were significantly more likely than males to receive prescriptions for: antibiotics; hormones; drugs affecting the central nervous, cardiovascular and urogenital systems; drugs for allergy and immune disorders; ear and nose topical preparations, and skin preparations, even after taking into account morbidity differences. If males and females were treated according to their presenting problems, differences in morbidity patterns would account for the management differences. However, the present investigation would suggest that GP and patient behaviours are also important factors that lead to differences in the prescriptions received by male and female patients in general practice. PMID- 9351149 TI - Health and gender differences between middle and senior managers in the Canadian Public Service. AB - Most studies of the relationship between socioeconomic status (SES) and health have concentrated on disparities between the richest and poorest men; few studies have examined such relationships for women due to difficulties in measuring SES for women. For the present study, data collected from Canadian Public Service middle and senior managers provided an opportunity to examine associations between SES and health within the upper end of the SES spectrum for both genders, since women managers can be assumed to have a relatively high SES. Demographic, health and lifestyle characteristics are compared for middle and senior managers for each gender separately to determine whether women experience the health benefits associated with higher SES that have been previously observed for men. The results support the hypothesis that achieving a higher SES through work is a more stressful process for women than for men and that women's upward mobility is restricted compared to that of men. Despite these findings, there is little evidence that women's health has been adversely affected. Compared to male managers, fewer female managers smoke or drink and fewer have high body mass index, high blood pressure or high cholesterol levels. Female managers are also more likely to report being in good health. PMID- 9351150 TI - "Culturalizing" the upsurge of tuberculosis. PMID- 9351151 TI - Thermometer use among Mexican immigrant mothers in California. AB - A community-based household survey was utilized to assess the relationship between thermometer use, home treatment and utilization of health care services. Using a cross-sectional design, the study surveyed 688 low income Mexican origin mothers of children between the ages of 8 and 16 months in San Diego County. Mothers were asked how they determine that their child has fever and how often they use a thermometer. Nearly 40% of low income Mexican mothers interviewed in San Diego county never used a thermometer for determining childhood fever. Approximately two-thirds (64.7%) relied either primarily or exclusively on embodied methods such as visual observation or touch to determine fever in their child. A multivariate logistic regression analysis determined that low education and a separated or divorced marital status decreased the odds of thermometer use, whereas regular contact with the health care system doubled the likelihood of thermometer use. Mothers who relied on embodied methods were more likely to use over-the-counter medications than those who relied on thermometers; however, no significant differences were found between groups using other methods of home treatment. Fever determination modalities can be used to screen for lack of access to care and to provide for other health care needs in a culturally appropriate manner. While clinicians' expectations may include parental experience with temperature taking, current pediatric literature questions the need for home-based thermometer use. Possible alternatives to the traditional rectal thermometer might include digital thermometers and color coded thermometer strips. PMID- 9351152 TI - The relationship between condition-specific morbidity, social support and material deprivation in pregnancy and early motherhood. ALSPAC Survey Team. Avon Longitudinal Study of Pregnancy and Childhood. AB - Poorer health has been consistently associated with both low levels of social support and material deprivation. It has been proposed that social support constitutes a causal link between health and deprivation such that those with lower socio-economic status have poorer health because their lack of social support makes them more vulnerable to disease. This assumption was tested in this study for women moving from pregnancy to early motherhood. The sample of 9208 women was drawn from the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC). Health status was measured by self-report of morbidity for three contrasting conditions: backache, depression and urinary infection. Data were collected by self-completion questionnaire at eight weeks prepartum and at eight weeks postpartum. The sample was divided into four groups for each condition on the basis of identification of the condition (1) on both occasions, (2) on neither occasion, (3) at eight weeks prepartum only, and (4) at eight weeks postpartum only. Chi-square tests were used to measure the association between presence or absence of a condition as defined above, material deprivation and low social support. Responses on an eight item social support questionnaire tapping emotional, instrumental and communal aspects of perceived social support were compared between these groups for each condition. Results showed significant associations, in late pregnancy and early motherhood, between poorer health and both material deprivation and low social support for two of the three conditions, depression and urinary infection. Changes in levels of perceived social support occurred as a consequence of motherhood; the percentage of women perceiving their partner to be supportive decreased as did the percentage of those who felt that their family or friends would help with financial difficulties; in contrast, the percentage of those perceiving that other mothers/neighbours offered support increased from late pregnancy to early motherhood. The extent and direction of such change was consistently associated with the presence or absence of depression, but not with backache or urinary infection. When mental health "improved", in that depression was present pre- but not postpartum, the percentage feeling supported increased and vice versa. This pattern was in evidence across all items of social support. It was concluded that perceived social support was unlikely in itself to constitute a causal link between poorer health and lower socio-economic status. In this context the relationship between social support and depression requires further consideration, particularly the extent to which they constitute component parts of a more general feeling of emotional well-being, a construct which is, in its turn, in need of further articulation in relation to health outcomes. PMID- 9351153 TI - Making sense of randomization; responses of parents of critically ill babies to random allocation of treatment in a clinical trial. AB - Randomized controlled trials (RCTs) are widely accepted by the scientific community as the most rigorous way of evaluating interventions in health care. Although their central feature, random allocation of treatment, is generally seen as methodologically appropriate, its application has caused much debate amongst health professionals and ethicists. This paper describes the views of parents who consented that their critically ill newborn baby should be enrolled in a neonatal trial. In-depth interviews were used to determine their response to the trial and randomization. The nature of the trial was often poorly understood. The random basis of the allocation of treatment and the rationale behind this approach were also problematic issues. Some parents did not perceive a random element in the process at all. These findings advance understanding of the perceptions of trial participants and raise important issues for those concerned with RCTs. Greater understanding of participants' views provides the potential to improve the management of future trials and so the experience of those agreeing to take part. PMID- 9351154 TI - Psychological and social predictors of motorcycle use by young adult males in New Zealand. AB - Motorcycle riding is a significant cause of serious injuries to young males. Little is known about the psychological and social characteristics of these riders, despite such knowledge being potentially important for the targeting of appropriate injury prevention interventions. Using problem-behaviour theory to broadly guide and structure the research, the present study focused on identifying predictors of motorcycle riding. Previous research investigating differences between riders and non-riders has tended to be inconclusive, methodologically limited, and lacking in explicit theoretical foundations. The present research was based on the birth cohort enrolled in the Dunedin Multidisciplinary Health and Development Study (DMHDS), a comprehensive New Zealand longitudinal study of health, development, attitudes, and behaviours. Logistic regression models were built using prior measures of health risk behaviour, other psychological and social factors, and motorcycle riding history as potential predictors of any motorcycle use at the age of 18 years. The strongest predictors were early motorcycle riding, including illegal on-road driving at age 13 (OR 4.0; 95% CI 1.7, 9.1), below average reading skills (OR 2.4; 95% CI 1.3, 4.6) and fighting in a public place at age 15 (OR 2.9; 95% CI 1.2, 6.9). It was of particular interest that this profile tended to fit less well those subgroups of riders with greatest exposure to on-road motorcycle driving. Although based on small numbers, this finding was consistent with earlier cross-sectional research that linked casual and unlicensed driving with less protective motorcycling opinions and behaviours. Some implications for injury prevention and public policies regarding motorcycling are discussed. In particular, stricter enforcement of present licensing regulations and stronger penalties for their violation could help to reduce the number of less responsible riders. PMID- 9351155 TI - Opportunities and impediments for a consolidating and expanding profession: genetic counseling in the United States. AB - Genetic counseling in the United States is consolidating and expanding its professional identity at a point in time when major biotechnological advances are rapidly being incorporated into clinical genetic practice and major shifts in the American health care system are being carried out under a mandate of cost containment. External factors--biomedical advances, political and economic contexts, managed care organizations, health economics criteria and exclusionary strategies of related health care occupations--combined with internal factors- training and certification requirements, specialization, leadership talent and ability to adapt to changing conditions--will strongly influence the structure and scope of genetic counseling services in the coming years. PMID- 9351156 TI - Marital status effects on health: are there differences between never married women and divorced and separated women? AB - To test whether the effects of marital status on health differ between never married women and divorced and separated women, this study utilizes prospective panel data for a large national sample of non-institutionalized young women in the U.S. (the National Longitudinal Surveys of Young Women). The women were aged 24-34 at the beginning of two successive five-year follow-up intervals (1978-1983 and 1983-1988). The health effects of marital status were evaluated in regressions which assessed the relationships between initial marital status and subsequent health trends in each follow-up interval. In the first follow-up interval, never married women tended to have worse health trends than divorced and separated women for physical impairments and for overall health problems. However, there were no differences between never married women and divorced and separated women in health trends for psychosomatic symptoms in either follow-up interval or for any health measure in the second follow-up interval. Our analyses of cross-sectional data showed few significant differences in health between never married women and divorced and separated women. Taken together, the evidence from our study and previous studies suggests that differences between never married women and divorced and separated women may vary by age and/or cohort. Evidence for the 1970s and 1980s suggests that, among older women, divorced and separated women may have experienced more harmful health effects than never married women; however, among younger women, this difference may have been absent or possibly reversed. PMID- 9351157 TI - Early probable Alzheimer's disease and awareness context theory. AB - The purpose of this research was to explore the explanatory value of Awareness Context Theory for social interactional issues in early probable Alzheimer's Disease (AD). Glaser and Strauss's Awareness Context Theory [Glaser and Strauss (1965) Awareness of Dying, Aldine, New York] served as the framework for the analysis of interview data from 14 early probable AD clients and 14 family caregivers, a written autobiographical account, a fictionalized account, observations of a family care-giver focus group, and excerpts that focused on early AD from field notes recorded during two years of participant observation at a specialized AD daycare center and a family caregiver support group. Initial open-ended study questions focused on the experience of early AD from the diverse perspectives represented in the data. After preliminary analysis of data suggesting emergent fit with Awareness Context Theory, questions were refocused to address awareness contexts. Data were coded and analyzed for fit with the theory. Awareness Context Theory provided a useful heuristic for thinking about the nuances and complexities of social interaction in early AD. Attention to awareness contexts should enable health care providers to suggest interventions to improve caregiver-client interactions. PMID- 9351158 TI - "We decide, you carry it out": a social network analysis of multidisciplinary long-term care teams. AB - The purpose of this study was to describe the structure of multidisciplinary long term care teams by identifying the pattern of relationships that develop amongst staff as they go about their work. Using a social network analysis approach, team members were classified as occupying the same structural position based on their patterns of relationships with other team members. The analysis was based on the results of a self-administered survey of 93 health care workers on three teams in the same multilevel geriatric care facility in Metropolitan Toronto. A common structure of the teams was identified consisting of two sub-teams: a multiprofessional sub-team and a nursing sub-team, each of which has a different structure indicating differential involvement in different types of teamwork. The multiprofessional sub-team has an "organic" structure and is mainly involved in teamwork that involves decision-making and problem-solving, whereas the nursing sub-team has a "mechanistic" structure and is mainly involved in task oriented work. The findings of this analysis indicate that while teamwork may be increasing the participation in decision-making by health professionals other than medicine, rather than flattening the hierarchical structure throughout the health care division of labour, its effects are limited to a group of higher status professionals. The clearly defined hierarchy remains for the lower status subdisciplines, and "I decide, you carry it out" has simply become "We decide, you carry it out". PMID- 9351159 TI - From careless consumptives to recalcitrant patients: the historical construction of noncompliance. AB - Thousands of articles on "noncompliance" have appeared since 1975. Yet the term has been criticized as paternalistic--as wrongly implying that patients should necessarily follow doctors' orders. This paper, which reviews how noncompliance has been constructed historically, argues that the problem with noncompliance is more than just one of terminology. Changing social and cultural factors during the 20th century have influenced the way in which uncooperative patients have been described. For example, resentment of poor immigrants in the early 1900s led doctors to describe tuberculosis patients who did not follow advice as "ignorant" and "vicious." Following World War II, patients who balked at taking new curative antibiotics for tuberculosis were called "recalcitrant." The term "noncompliance," popularized by Sackett and Haynes in the 1970s, reflected their early role in the field of research now termed "evidence-based medicine." While Sackett and Haynes had hoped that the new term would eschew earlier value judgments, noncompliance, through its association with the positivistic ethos of evidence-based medicine, has been conceptualized as a "tragic" problem potentially solvable by clinical research. Hence, noncompliant patients are still seen as deviant. With the growth of managed care in the United States, there is increasing pressure to get patients to follow medical recommendations. History suggests that labels such as "noncompliant" are invariably judgmental. Rather than seeing the provider's role as trying to get noncompliant patients to comply, we should emphasize the importance of negotiation and accommodation within the provider-patient relationship. PMID- 9351160 TI - The effects of the cultural context of health care on treatment of and response to chronic pain and illness. AB - Qualitative data from two studies in Puerto Rico and New England are used to show how cultural values, standards and beliefs in different health care contexts affect (1) health care professionals' responses to patients' problems, (2) the relationships between providers and patients, and (3) the patients' responses to chronic pain and illness. Influencing elements in the care setting include the world view of the relationship of mind and body in illness processes, the dominant values and standards regarding pain and illness behaviors and the degree of cooperation between the providers and other agencies the patient depends on for compensation, rehabilitation and health insurance. In the New England study, the biomedical world view of mind-body dualism was shared by providers and most patients, but this shared belief often contributed to substantial patient stress and alienation. In contrast, in the Puerto Rican study providers and patients often shared a view of mind-body integration in illness and valued treatments which addressed chronic pain as a biopsychosocial experience. In this setting, shared views and values contributed to more supportive patient-provider relationships, and patients thus experienced less treatment-related stress. PMID- 9351161 TI - The significance of the mouth in old age. AB - Information on the significance of the mouth in old age has been obtained from structured interviews with older subjects focused largely on the significance and impact of oral dysfunction. There is, however, a growing sense that inventories of dysfunction do not explain the full significance of aging and that structured interviews offer little opportunity to explore feelings and concerns. This study adopted a qualitative approach to collect and analyse data from unrestricted responses to the question: "What is the significance of oral health in the lives of older adults?" The data were collected by interviewing 24 elders, and major themes in transcripts of the interviews were identified by the research team using inductive analytical techniques. Our findings indicate that the significance of oral health in this age group was considered largely within the context of three interacting themes--comfort, hygiene and health--that can be illustrated within a theoretical framework that corresponds with more general theories of aging to offer guidance for health promotion and further research. Overall, the participants offered a positive perspective on the mouth, and they emphasized the need to adapt as an integral part of successful aging and a means of coping with the impact of oral disorders. PMID- 9351162 TI - Muscle inhibition following knee injury and disease. AB - It has been observed that knee extensor muscles cannot be fully activated during voluntary contractions following knee injuries. This muscle inhibition has an unknown origin and appears to hinder full rehabilitation of the affected joint. We have investigated muscle inhibition during and following knee injuries in non athletic and athletic patients and compared their results to non-athletic, unaffected volunteer subjects. There appears to be a small amount of muscle inhibition in the knee extensors of normal subjects; this inhibition increases dramatically following knee injury, and appears to go back to normal levels following surgical intervention, aggressive physiotherapy, or a sufficient amount of time. Depending on the intervention, strength deficits of the affected compared to the unaffected knee extensor muscles may persist. Aggressive physiotherapy can eliminate strength deficits following knee injury through an increased ability to recruit the knee extensors in patients more completely compared to normal subjects. PMID- 9351163 TI - The relationship between electromyogram and muscle force. AB - Some guidelines are given for the interpretation of the electromyogram (EMG). In static isometric contractions there is, usually a linear, relationship between muscle force and smoothed rectified EMG (SRE). It should be considered, however, that most of the time several muscles are active simultaneously around a joint. This finding implies that the relationship between the SRE of a single muscle and the total joint moment needs not be linear. During movements, the force-length velocity relationship and the elastic properties of muscle should be taken into account. Also, EMGs during concentric movements are larger than those measured during isometric actions. A further consideration in relating force to EMG is that the force signal has a much lower frequency content than the rectified EMG. Finally, the limited speed of the activation and deactivation process results in a 50 to 200 ms delay of the muscle force relative to the EMG. PMID- 9351164 TI - Sizing clinical trials with variable endpoint event rates. AB - Although many researchers in cardiovascular clinical trials have disciplined themselves to execute sample size calculations in the design of their studies, these computations become difficult in the presence of control group endpoint event rate uncertainty. Recent experience in cardiovascular clinical trials suggests that, although one may know the control group event rate during the design phase of the trial, it can decrease during the trial's execution. Its resultant overestimation can lead to a power reduction with serious consequences for the trial's interpretation. Although the investigators may acknowledge the likelihood that the control group event rate will decrease during the time course of the trial, there is no formal means to adjust the design phase estimate. In this paper, I first formulate the sample size as a function of the control group event rate theta and then I place a proper probability distribution on theta, allowing for the uncertainty in this parameter's value during the course of the study. From this assumption, the sample size itself becomes a random variable, whose expectation and variance are computed. I explore the implications for sample size for various reasonable proper probability distributions on the control group event rate. PMID- 9351166 TI - A simple non-linear model in incidence prediction. AB - A simple model is proposed for incidence prediction. The model is non-linear in parameters but linear in time, following models in environmental cancer epidemiology. Assuming a Poisson distribution for the age and period specific numbers of incident cases approximate confidence and prediction intervals are calculated. The major advantage of this model over current models is that age specific predictions can be made with greater accuracy. The model also preserves in the period of prediction the age pattern of incidence rates existing in the data. It may be fitted with any package which includes an iteratively reweighted least squares algorithm, for example GLIM. Cancer incidence predictions for the Stockholm-Gotland Oncological Region in Sweden are presented as an example. PMID- 9351165 TI - The latent class model for multiple binary screening tests. AB - Given multiple binary tests, such as repeated application of a blind screening test to each individual in a sample, we attempt to estimate the prevalence, sensitivity and specificity of the test without knowing the true disease status of those tested (gold standard). This problem is equivalent to finding the mixing distribution of a mixture of binomial distributions. We suggest a new method to determine the number of latent classes. Our simulations show that the coverage probabilities of the bootstrap confidence intervals of our estimates are correct. Our methods are illustrated by examples from published medical research. PMID- 9351167 TI - Hierarchical polytomous regression models with applications to health services research. AB - The analysis of variations is an important area of interest in health services and outcomes research and has two main goals: to identify and quantify variability across units, such as geographic regions or health care providers, in terms of procedure utilization and outcomes, and to explore the links between process, such as regional or hospital practice patterns, and outcomes, such as patient mortality and functional status. Hierarchical regression models are well suited for this type of analysis. In this paper we formulate a hierarchical polytomous regression model and apply it to the analysis of variations in the utilization of alternative cardiac procedures in a national cohort of elderly Medicare patients who had an acute myocardial infarction during 1987. The model is designed to accommodate clustered multinomial data with covariate vectors available on individual cases and on clusters. We present a Bayesian approach to fitting and checking the model using simulated values from the posterior distribution of the parameters. The simulation algorithms are based on Gibbs sampling in combination with Metropolis steps. Using the hierarchical polytomous regression model, we examine how the rates of cardiac procedures depend on patient-level characteristics, including age, gender and race, and whether there exist interstate differences and regional patterns in the use of these procedures. PMID- 9351168 TI - A method for deciding early stopping of inconclusive case-control studies in settings where data are stratified. AB - Statistical methodology for early stopping of clinical trials is well developed. However, in epidemiologic studies, methods for deciding early stopping have rarely been considered. In contrast to clinical trials, termination due to an early significant result is indeed seldom relevant, because further expenditure is then likely to be worthwhile in order to obtain a more precise effect estimate and to more precisely account for confounders. On the other hand, if data obtained early indicate an inconclusive final result and if continuation of the epidemiologic study is expensive, then it might be desirable to stop the study early. The present paper proposes a method for evaluating the possibility that an ongoing case-control study will produce an inconclusive final result. The method is developed for stratified case-control data. In principle, probable final results, given the available data, are predicted by repeatedly simulating the remaining data using plausible assumptions on the true exposure effect. These assumptions involve the current effect estimate as well as some alternatives, chosen with respect to the investigator's prior hypothesis concerning a harmful exposure effect, which are in reasonable agreement with the available data. PMID- 9351169 TI - Estimating a delay distribution from incomplete data, with application to reporting lags for AIDS cases. AB - Statistical inference for the probability distribution of a reporting delay is considered when delays are recorded only after a certain point in time tau. A method is proposed which utilizes data on incidences arising prior to tau. By a suitable choice of parameters we find explicit expressions for maximum likelihood estimates and standard errors. An application to reporting delay data on Australian AIDS diagnoses demonstrates that inclusion of data on AIDS diagnoses made prior to tau results in significant gains in the precision of estimates. A simulation study indicates for this data set that there is minimal bias in the estimates and the large sample formulae for the standard errors give good estimates of the standard deviation of the estimators. PMID- 9351171 TI - New strategies for antibacterial drug design. PMID- 9351170 TI - Using the general linear mixed model to analyse unbalanced repeated measures and longitudinal data. AB - The general linear mixed model provides a useful approach for analysing a wide variety of data structures which practising statisticians often encounter. Two such data structures which can be problematic to analyse are unbalanced repeated measures data and longitudinal data. Owing to recent advances in methods and software, the mixed model analysis is now readily available to data analysts. The model is similar in many respects to ordinary multiple regression, but because it allows correlation between the observations, it requires additional work to specify models and to assess goodness-of-fit. The extra complexity involved is compensated for by the additional flexibility it provides in model fitting. The purpose of this tutorial is to provide readers with a sufficient introduction to the theory to understand the method and a more extensive discussion of model fitting and checking in order to provide guidelines for its use. We provide two detailed case studies, one a clinical trial with repeated measures and dropouts, and one an epidemiological survey with longitudinal follow-up. PMID- 9351172 TI - Mycobacterium tuberculosis: bringing down the wall. PMID- 9351173 TI - The role of superantigens in resistance to retroviral infection. PMID- 9351174 TI - Origins of the mobile gene cassettes found in integrons. AB - Many of the acquired antibiotic resistance genes found in enterobacteria and pseudomonads are part of small mobile elements known as gene cassettes, and other genes are also likely to be found in cassettes. The origins of the genes and the recombination sites that make up cassettes are not known, but recent analyses of available data suggest that cassettes may be ancient structures, and some hypotheses for how they are formed can now be examined. PMID- 9351175 TI - Bacterial avirulence proteins as triggers of plant disease resistance. AB - Disease resistance in plants is often characterized by matching resistance and avirulence genes in host and pathogen, respectively. It has recently been shown that expression of bacterial avirulence genes in plants induces resistance gene dependent defense reactions. The finding that avirulence proteins act inside plant cells represents a major advance in our understanding of host-pathogen specificity. PMID- 9351176 TI - Reactivation of Epstein-Barr virus: regulation and function of the BZLF1 gene. AB - The switch from latent infection to virus replication in Epstein-Barr virus (EBV) infected B cells is initiated by expression of the viral BZLF1 gene. Recent studies have identified the key cellular transcription factors involved in regulating this switch in viral programs and the signal transduction pathways to which they respond. Understanding this switch may facilitate development of strategies to interfere with EBV infection. PMID- 9351177 TI - Hydrophobins and fungal infection of plants and animals. AB - Hydrophobins are among the most important structural proteins produced by fungi. Their least-understood function is how they act to promote infection-related morphogenesis. Although the hydrophobin of at least one plant pathogen appears to be involved as a signal molecule in pathogenesis, a role for hydrophobins in animal pathogenesis has not been convincingly documented. PMID- 9351178 TI - Pathogenesis of dengue virus diseases: missing pieces in the jigsaw. AB - The mechanisms involved in the pathogenesis of dengue hemorrhagic fever and dengue shock syndrome remain unresolved. Antibody-dependent enhancement of infection has long been thought to play a central role; however, this remains unverified. The alternative hypothesis that virus variation, virulence and dynamics may account for severe dengue disease, particularly in children, should be considered. PMID- 9351179 TI - Mycoplasma genes: a case for reflective annotation. AB - Although function can be assigned to genome sequence by homology at a macroscopic level, this can be misleading in the absence of data on enzyme activities. Together, such data can reveal whether open reading frames are expressed, identify multienzyme function and point to 'orphan' function. Because of their small size and small genomes, the genome sequences of some Mycoplasma spp. are very amenable to detailed analyses. PMID- 9351180 TI - Food safety: the veterinarian's central role. PMID- 9351181 TI - Abnormalities of heart rate and rhythm in bovine spongiform encephalopathy. AB - Heart rates of healthy cows and cows suspected of having bovine spongiform encephalopathy were measured by auscultation and by a portable cardiac monitor. Bradycardia was demonstrated in suspect cases which were confirmed histopathologically. Disturbances in cardiac rhythm were also evident in some cases. Healthy cows deprived of food exhibited bradycardia. The administration of pharmacological doses of atropine indicated that bradycardia in BSE was mediated by increased vagal influence, suggesting that the cardioinhibitory reflexes in the caudal brainstem were functionally altered by the disease. PMID- 9351182 TI - Efficacy of moxidectin, ivermectin and albendazole oral drenches for suppression of periparturient rise in ewe worm egg output and reduction of anthelmintic treatment for lambs. AB - Sixty multiparous crossbred ewes which had lambed within three days in the first week of April 1996, were divided into four groups. Each group consisted of 15 ewes plus 12 pairs of twins and three single lambs. Group 1 was left untreated, group 2 was treated with albendazole 2.5 per cent drench, group 3 received moxidectin 0.1 per cent drench and group 4 received ivermectin 0.08 per cent drench. The ewes in each group were dosed with their anthelmintic on April 4 (day 0) before being turned out to separate equal-sized paddocks within the same field on the following morning. The field had been used for grazing sheep annually for many years and was considered to be contaminated with infective larvae of the common gastrointestinal nematodes infecting sheep in the region. Faecal samples were collected every two weeks from the ewes and lambs until July 25 (day 112). The lambs in each group were dosed with the anthelmintic used for their dams on day 42, and the dose was repeated when more than 50 per cent of the lambs in any group had a faecal egg count of more than 200 eggs per gram (epg). The total faecal egg output of the treated ewes over days 14 to 70, compared with that of the untreated control group, was reduced by 78.9 per cent by the moxidectin drench, by 47.6 per cent by ivermectin, and by 21.5 per cent by albendazole. The lambs in the groups treated with moxidectin and ivermectin required only one treatment on day 42 before reaching finishing weight; those in the albendazole treated group were treated twice and the control group once. The faecal egg outputs of the lambs from day 42 until the end of the experiment on day 112 were reduced by 75 per cent by the moxidectin drench, by 48.5 per cent by ivermectin, and by 9 per cent by albendazole. There were no significant differences between the rates of weight change of either ewes or lambs in any of the groups. PMID- 9351183 TI - Canine monocytic ehrlichiosis: a retrospective study of 100 cases, and an epidemiological investigation of prognostic indicators for the disease. AB - One hundred cases of monocytic ehrlichiosis diagnosed in Israeli dogs were confirmed by the presence of anti-Ehrlichia canis indirect immunofluorescent antibody titres greater than 1:40. The disease occurred in all age groups and there was no sex predilection. German shepherd dogs were significantly over represented whereas crossbreed dogs were significantly under-represented (P > 0.0005). The most common clinical signs were depression, lethargy, lymphadenomegaly, fever, anorexia, panting, pale mucous membranes and bleeding, of which epistaxis was most common. Thrombocytopenia, anaemia (mainly normocytic normochromic) and lymphopenia were the predominant haematological findings. Forty nine of the 100 cases were followed up for a year. Thirty-two dogs survived and 17 died. A Cox proportional hazards regression model was used to examine the effect of host, environmental, and haematological prognostic factors on survival. It was concluded that severe anaemia, severe leucopenia, pancytopenia, a tendency to bleed (especially epistaxis) and being a German shepherd dog were important indicators of poor survival in cases of monocytic ehrlichiosis in dogs. PMID- 9351184 TI - Effect of time between farm loading and processing on carcase quality of broiler chickens. PMID- 9351185 TI - Successful cryopreservation of porcine embryos by vitrification. PMID- 9351186 TI - Farm assurance schemes. PMID- 9351187 TI - Chronic hepatopathy (hepatic lipodystrophy) of Galloway cattle. PMID- 9351188 TI - Structure and thermal vibrations of spermine phosphate hexahydrate from neutron diffraction data at 125 K. AB - Spermine phosphate hexahydrate crystallizes in space group P2(1)/a with unit-cell dimensions a = 7.931 (1), b = 23.158 (5), c = 6.856 (2) A, and beta = 113.44 (2) degrees at 125 K with unit-cell contents [(C10H30N4)2(4+)(HPO4)4(2-).12H2O]. The packing of spermines and monohydrogen phosphates in this crystal structure has features which may be relevant to the binding of spermine to DNA. Another important structural feature is the presence of channels containing water that is hydrogen bonded as in ice-Ih with disordered protons. The channels occur between sheets of spermine long chains and are also bordered by hydrogen-bonded monohydrogen phosphate chains. The hydrogen-bonding scheme of these water chains proposed on the basis of an earlier X-ray study is now confirmed. Nuclear positions, anisotropic mean-square (m.s.) displacements, an overall scale factor and two extinction parameters (rho and g) were refined using full-matrix least squares giving values of R(F0(2)) = 0.09, Rw(F0(2)) = 0.11 and S = 1.02. Thermal vibrational analysis revealed that the backbone of the spermine cation can be described as a single rigid segment with a substantial libration of 27 deg2 around the spermine molecular long axis. PMID- 9351189 TI - The importance of intermediate filaments in the adaptation of tissues to mechanical stress: evidence from gene knockout studies. AB - Research over the past few years on the function of intermediate filaments in cells in culture has not produced convincing results, because the key role of intermediate filaments is within tissues and at certain periods of development. Only recently the technique of gene knockout has been used to examine intermediate filaments in mice and has provided the first evidence that intermediate filaments are directly involved in cell resilience and the maintenance of tissue integrity. Knockout of the gene encoding keratin K8 is lethal in the embryo, and results in hepatic or intestinal lesions, while knockout of the K14 or K10 genes leads to rupture of stratified epithelia. Knockout of the gene encoding desmin causes the rupture of skeletal and cardiac muscle, and collapse of blood vessel walls. Knockout of the gene coding for GFAP leads to a loss of cerebral white matter, and knockout of the gene coding for vimentin causes degeneration of the cerebellar Purkinje cells. The results reveal the lack of compensation by another intermediate filament. Tissues without intermediate filaments fall apart; they are mechanically unstable, unable to resist physical stress, and this leads to cell degeneration. By maintaining the shape and plasticity of the cell, the intermediate filament network acts as an integrator within the cell space. The state of mechanical force imposed on a tissue or a cell can alter the shape of certain elements of the cytoskeleton and thus participate to the control of cell functions. PMID- 9351190 TI - TPA induces apoptosis in MPC-11 mouse plasmacytoma cells grown in serum-free medium. AB - Apoptotic-like events could be rapidly induced by the phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA) in cells of the mouse plasmacytoma cell line MPC-11 grown in serum-free medium. Indicators for apoptosis were morphological changes visualized by light and electron microscopy, such as chromatin condensation and the formation of cellular buds and fragments, as well as biochemical indices like the appearance of the so-called 'DNA ladder'. Additionally, in these cells which are usually devoid of significant amounts of cytoplasmic intermediate filament (cIF) proteins, synthesis and accumulation of the cIF protein vimentin was rapidly induced by TPA treatment and almost all cells became vimentin-positive. Later on, substantial amounts of vimentin and lamin B degradation products appeared, and an increasing fraction of cells displayed low or even undetectable quantities of intact vimentin. This subpopulation was characterized via microscopy to be in the late stages of apoptosis. We suggest that in MPC-11 cells undergoing apoptosis in response to TPA treatment vimentin as well as lamin B are degraded, leading to a rearrangement and eventual loss of their respective filament networks. PMID- 9351191 TI - Antisense inhibition of beta-actin mRNA localization and its effect on smooth muscle cell migration. AB - A crucial step in cell migration involves changes in the actin cytoskeleton in response to extracellular signals. We have previously shown that beta-actin transcripts are associated with mobile regions of mouse 3T3 fibroblasts when grown in the presence of serum. In the current study we used in situ hybridization and laser scanning confocal microscopy to show that cultured rat smooth muscle cells also localize beta-actin mRNA to the cell periphery and that this peripheral pool of beta-actin mRNA is dependent on the presence of growth factors in the culture medium. We also show that antisense phosphorothioated oligonucleotides directed against sequences in the 3' untranslated region of rat beta-actin mRNA block peripheral localization of beta-actin mRNA while the corresponding control oligonucleotides have no effect. Time-lapse video analysis demonstrates that the antisense oligonucleotides inhibit rat smooth muscle cell migration in culture and analysis of beta-actin mRNA confirms this is not due to changes in beta-actin gene expression or instability of the message. Our results suggest that depletion of beta-actin transcripts from the cell periphery is associated with suppression of SMC migration. PMID- 9351192 TI - Ultrastructural changes in the Golgi apparatus and secretory granules of HL-60 cells treated with the imino sugar N-butyldeoxynojirimycin. AB - The imino sugar N-butyldeoxynojirimycin inhibits the N-linked oligosaccharide processing enzymes alpha-glucosidases I and II, and the ceramide specific glucosyltransferase which catalyses the first step in glucosphingolipid biosynthesis. We have studied the effects of this compound on the ultrastructure of HL-60 cells to identify novel activities of this compound. Treatment of HL-60 cells with this imino sugar results in several morphological changes within the cell, none of which result in cytotoxicity. The plasma membrane stains heavily with potassium ferrocyanide within 30 min following addition of the compound to the medium, and there is then a time dependent involvement of all other intracellular membranes. Secretory granules become enlarged and lose their dense core morphology and appear either empty and vacuolated or have low density contents. However, the most striking effect of NB-DNJ treatment is on the Golgi apparatus. The Golgi exhibits a time-dependent change from typical Golgi morphology to a structure almost completely devoid of cisternae and consisting predominantly of vesicles. All the observed changes are fully reversible on withdrawal of the compound. PMID- 9351193 TI - Lectin binding pattern and band 3 localization in toad skin epithelium and the effect of salt acclimation. AB - Seven lectins were employed to localize glycoconjugates in the skin of a toad (Bufo viridis). Each of the lectins exhibited a particular, specific and selective binding pattern. Peanut lectin (PNA) and WGA bound to mitochondria-rich (MR) cells, but WGA bound also abundantly, in the dermis. Band 3-like protein, as indicated by the reaction with polyclonal anti band 3 antibody, was localized exclusively in MR cells. Ionic acclimation (200 mmol/L NaCl, or 50 mmol/L KCl) affected profoundly the binding pattern of the lectins. High NaCl acclimation resulted also in diminishing anti band 3 antibody binding, whereas in skins of KCl-acclimated toads the staining remained similar to the control. The binding of WGA but not PNA, corresponded with the same cells that stained with anti band 3 antibody. PNA in concentration of > 10 micrograms/mL reduced reversibly, both the resting and activated Cl- conductance by 25-30%. Based on differential binding of band 3, WGA and PNA, these observations provide conclusive verification of the presence of at least two populations of MR cells in the toad skin epithelium. It is suggested that the PNA positive MR cells may correspond to a beta-type MR cell. The information can be used to study molecular mechanisms that are involved in ionic acclimation. PMID- 9351194 TI - Evolution of the hydrogenosome. AB - Since its discovery almost 25 years ago the enigmatic hydrogenosome, a redox organelle of anaerobic unicellular eukaryotes, has puzzled evolutionists as to its origin and function. Synthesis of recent molecular, physiological and morphological studies now favours the hypothesis that hydrogenosomes derived from a modification of pre-existing mitochondria, and argues against the previously held view that the hydrogenosome had a polyphyletic origin. These data provide evidence for a more ancient origin of mitochondria than hitherto thought. PMID- 9351195 TI - Trans-acting regulation of antibiotic TA genes in Myxococcus xanthus. AB - Two regulatory mutations of Myxococcus xanthus, which cause an increase in the transcription of genes required for antibiotic TA synthesis, were mapped by transduction and their effect on transcription of four TA genes examined. The two regulatory mutations were closely linked and located within the 40-kb TA gene cluster on the M. xanthus chromosome. Recombinants were constructed which contained one of the regulatory mutations and promoter probes in the four different TA genes. Both regulatory mutations enhanced transcription of three of the four TA genes. However, construction of a strain containing the over expression regulatory mutation in a wild-type background produced less antibiotic than the parental strain. PMID- 9351196 TI - Degradation of the polyamine alkaloid aphelandrine by endophytic fungi isolated from Aphelandra tetragona. AB - Members of the genus Aphelandra (Acanthaceae) produce rare macrocyclic polyamine alkaloids which consist of spermine acylated with two units of 3-(4 hydroxyphenyl)prop-2-enoic acid. Endophytic fungi were isolated from roots and shoots of Aphelandra tetragona and tested for their ability to metabolize the main alkaloid aphelandrine, which accumulates exclusively in the roots of the plants. Several endophytes were able to metabolize aphelandrine but only root endophytes belonging to the Nectriaceae were good metabolizers. In addition, the endophytes were grown on an agar medium containing putrescine, spermidine, or spermine as the sole nitrogen source. All fungi were able to grow on putrescine, but only the good aphelandrine metabolizers grew well on spermidine or spermine. Acremonium sp. 15, one of the most active metabolizers, grew also on a medium containing aphelandrine as sole nitrogen source. A number of strains thought to be conspecific with Acremonium sp. 15 were also tested for their ability to metabolize aphelandrine. The ability of the endophytes to metabolize aphelandrine suggests an ecological adaptation of the symbionts to their host. The possibility of using the aphelandrine metabolism as a taxonomic character is briefly discussed. PMID- 9351197 TI - The biodegradation of tributyl phosphate by naturally occurring microbial isolates. AB - The biodegradation of tributyl phosphate by a mixed culture of Pseudomonads was demonstrated. Growth and the rate of tributyl phosphate consumption were variable and divisible into rapid and slow rates. Rapidly growing, rapidly tributyl phosphate-utilising cultures contained a 22-24 kb DNA fragment isolated by two methods, which was not visible in the cultures growing slowly. The mixed culture gave five periods of rapid growth interspersed with periods of poor growth during 7 months of weekly subculture, with the 22-24 kb DNA fragment detectable during the rapidly growing periods only. Seven Pseudomonads isolated from the culture grew at the expense of tributyl phosphate as the sole phosphorus source but spontaneously and irreversibly lost this ability after eight serial subcultures. PMID- 9351198 TI - Comparison of two PCR methods for rapid identification of Leptospira genospecies interrogans. AB - Based on (i) an analysis of Leptospira 16S rDNA sequences determined by us and of those from databases and (ii) a previously published finding that restriction fragment length polymorphisms (RFLPs) within the Leptospira 16S and 23S rDNA were detected by nine restriction enzymes and these RFLPs allowed categorisation of Leptospira into eight genospecies, we predicted that one particular DdeI restriction site polymorphism within 16S rDNA could be independently used for identifications of Leptospira strains belonging to the genospecies interrogans. Two PCR-based methods, namely allele-specific amplification (ASA) and PCR-RFLP, were tested for the rapid detection of the DdeI restriction site polymorphism. One or two representative strains from each of nine genospecies were tested by ASA, whereas 73 strains from nine genospecies and two field isolates were tested by PCR-RFLP. Our experiments showed that the ASA method was not as specific as intended, but the PCR-RFLP method was useful for rapid identifications of the genospecies interrogans. We have not only confirmed a previous finding and extended the number of samples particularly from the genospecies biflexa, weilii, and inadai, but also simplified a previous PCR-RFLP protocol. PMID- 9351199 TI - Bacterial flavohaemoglobins: a consensus sequence and identification of a discrete enterobacterial group and of further bacterial globins. AB - The amino acid sequences of haemoglobin-like proteins from the bacteria Alcaligenes eutrophus, Bacillus subtilis, Erwinia chrysanthemi, Escherichia coli, Vibrio parahaemolyticus, Vitreoscilla sp. and the yeast Saccharomyces cerevisiae were studied. Phylogenies based on distance and parsimony analysis showed that the eubacterial group can be easily distinguished from the other haemoglobin-like proteins. The construction of a consensus bacterial flavohaemoglobin based on the alignment of six bacterial and one yeast globins allowed the design of consensus primers to search for haemoglobin-like genes in other bacteria. PCR products of the expected size were found in Campylobacter jejuni, Salmonella typhimurium, Listeria monocytogenes, Rhizobium leguminosarum, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. PMID- 9351200 TI - Spirochaeta smaragdinae sp. nov., a new mesophilic strictly anaerobic spirochete from an oil field. AB - An obligately anaerobic spirochete designated strain SEBR 4228T (T = type strain) was isolated from an oil field of Congo, Central Africa. The strain grew optimally with a sodium chloride concentration of 5% (sodium chloride concentration) growth range 1.0-10%) at 37 degrees C (growth temperature range 20 40 degrees C) and pH of 7.0-7.2 (pH growth range pH 5.5-8.0). Strain SEBR 4228T grew on carbohydrates (glucose, fructose, ribose, D-xylose, galactose, mannitol and mannose), glycerol, fumarate, peptides and yeast extract. Yeast extract was required for growth and could not be replaced by vitamins. It reduced thiosulfate and sulfur, to H2S. Glucose was oxidised to lactate, acetate, CO2 and H2S in the presence of thiosulfate but in its absence lactate, ethanol, CO2 and H2 were produced. Fumarate was fermented to acetate and succinate. The G + C content of strain SEBR 4228T was 50%. Strain SEBR 4228T was spiral shaped measuring 5-30 by 0.3-0.5 micron and was motile with a corkscrew-like motion. Electron microscopy revealed the presence of periplasmic flagella in a 1-2-1 arrangement. Strain SEBR 4228T possessed features typical of the members of the genus Spirochaeta. 16S rRNA sequence analysis revealed that it was closely related to Spirochaeta bajacaliforniensis (similarity 98.6%). The lack of DNA homology with S. bajacaliforniensis (38%), together with other phenotypic differences, indicated that strain SEBR 4228T is a new species, which we have designated Spirochaeta smaragdinae. The type strain is SEBR 4228T (= DSM 11293). PMID- 9351201 TI - Nickel resistance in Escherichia coli V38 is dependent on the concentration used for induction. AB - Strain Escherichia coli V38 resistant to 4 mM NiCl2 was isolated from the city sewage sludge. It showed low nickel accumulation by cells and nickel ion efflux. Cells were pregrown (induced) overnight in the presence of Ni2+, then the culture was kept on ice for 20-30 min and transferred to 37 degrees C for further incubation. When the Ni2+ concentration during growth was the same as during incubation, there was no noticeable accumulation of Ni2+. When the Ni2+ concentration during incubation was higher than that used for induction, uptake of 63Ni2+ and delayed efflux were seen. The uptake and delay of both efflux and growth were directly proportional to the difference between the concentrations used for induction and incubation. Active nickel ion uptake was seen in cells taken from cultures in the delayed efflux period. PMID- 9351202 TI - Protection against haemorrhagic septicaemia induced by vaccination of buffalo calves with an improved oil adjuvant vaccine. AB - An experimental oil adjuvant vaccine was developed against haemorrhagic septicaemia, a disease of cattle and buffalo caused by Pasteurella multocida serotype B and E. Mineral oil, Mercol 52, was used as adjuvant together with Span 85 and Tween 85 as emulsifiers. The vaccine was evaluated by single dose intramuscular immunisation of 1-2 year old buffalo calves. IgG and IgM class antibodies were determined by ELISA. The group of animals immunised with the experimental oil adjuvant vaccine showed a high titre of the IgG class of antibodies measured at 300 days post vaccination. To compare the protective efficacy of the vaccine with the commonly used broth bacterin, another group of buffalo calves was immunised by broth bacterin. This group showed a low level of IgG antibodies. Protection was assessed by challenge with 10(9) viable bacteria of P. multocida type B:2,5 administered subcutaneously, 250 days post vaccination. Animals vaccinated with the experimental oil adjuvant vaccine were fully protected. The other groups of animals, vaccinated with broth bacterin or used as control (non-vaccinated), developed symptoms of haemorrhagic septicaemia. A strong relationship between IgG but not IgM class antibody level and resistance to challenge was observed. The experiment demonstrated that the experimental oil adjuvant vaccine was superior to broth bacterin in providing protection against experimental haemorrhagic septicaemia in young buffalo calves beyond 250 days. PMID- 9351203 TI - Pyroglutamic acid and iron regulate the expression of the pcp gene in Pseudomonas fluorescens MFO. AB - Pyrrolidone carboxyl peptidase (Pcp) is an aminopeptidase (EC 3.4.11.8) able to specifically remove the L-pyroglutamyl residue from the amino-terminus of polypeptides. Since nothing was known concerning the regulation and function of Pcps, a mutant of a milk-isolated strain lacking Pcp activity (Pseudomonas fluorescens MB1), was constructed by homologous recombination using a transcriptional fusion between pcp and a reporter gene (uidA). The wild-type and mutant strains were grown in synthetic media and in milk to investigate the environmental effects on pcp transcription. The expression of pcp and of the transcriptional fusion pcp::uidA was not sensitive to environmental conditions like temperature, osmolarity or nitrogen and phosphate starvation but was induced by the product of the enzymatic activity, pyroglutamic acid (pGlu). The expression of the native gene and the fusion in inducing conditions was also controlled by the iron concentration. The identification in the pcp promoter sequence of putative ferric uptake regulator (Fur) binding sites suggests a transcriptional regulation in a Fur-dependent fashion. Two other putative regulatory stretches, corresponding to inverted repeated sequences with perfect and imperfect symmetry, were also identified. pGlu and iron are therefore at least two of the transcriptional effectors of pcp expression. PMID- 9351204 TI - Sequence analysis of a 1296-nucleotide plasmid from Xylella fastidiosa. AB - A cryptic plasmid from Xylella fastidiosa strain ATCC 35868 was cloned, sequenced, and the sequence entered into GenBank (U71220). The plasmid is 1296 nucleotides in length with 55% GC content and three open reading frames. A plasmid with sequence homology was found in only one other strain of X. fastidiosa, ATCC 35878. Searches of the GenBank reveal nucleotide sequence homology with plasmid pNKH43 from Stenotrophomonas maltophilia, and amino acid sequence homology with phage Pf3 from Pseudomonas aeruginosa, plasmid pAP12875 from Acetobacter pasteurianus, and plasmid pVT736-1 from Actinobacillus actinomycetemcomitans. PMID- 9351205 TI - High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes. AB - Many streptomycetes, including S. coelicolor A3(2), possess a potent methyl specific restriction which can present an effective barrier to the introduction of heterologous DNA. We have compared the efficiency of intergeneric conjugal transfer of different types of plasmids to S. coelicolor and S. lividans 66 using two E. coli donors: the standard, methylation proficient strain S17-1, and the methylation deficient donor, ET12567(pUB307). We demonstrate that the methylation deficient donor can yield > 10(4)-fold more S. coelicolor exconjugants than the standard donor. In the case of pSET152 derivatives, which integrate into the host chromosome by site-specific recombination, up to 10% of streptomycete spores in the conjugation mixture inherit the plasmid. The conjugation procedure is efficient enough to obtain exconjugants with 'suicide' delivery plasmids and therefore provides a simple route for conducting gene disruptions in methyl DNA restricting streptomycetes, and possibly other bacteria. PMID- 9351206 TI - Note: purification of amylase secreted from Bifidobacterium adolescentis. AB - Bifidobacterium adolescentis Int-57 isolated from human faeces produced extracellular amylase. The enzyme was purified from the culture supernatant fluids by ammonium sulphate precipitation, gel-filtration chromatography (Sephadex-G-75), ion-exchange chromatography (CM-cellulose) and FPLC. SDS-PAGE of the purified enzyme revealed a major band with an apparent molecular weight of 66 kDa. The pI was 5.2. Enzyme activity was optimal at 50 degrees C, and at pH 5.5. The enzyme was stable at 20-40 degrees C, and at pH 5-6 with a K(m) value of 2.4 g l-1 soluble starch. The activation energy was 42.3 kJ mol-1. The enzyme was significantly inhibited by maltose (10%), glucose (10%), Cu2+ (5 mmol l-1), Zn2+ (5 mmol l-1), N-bromosuccinimide (5 mmol l-1), EDTA (5 mmol l-1), I2 (1 mmol l-1) and activated by beta-mercaptoethanol (10 mmol l-1). PMID- 9351207 TI - The germinability of spores of a psychrotolerant, non-proteolytic strain of Clostridium botulinum is influenced by their formation and storage temperature. AB - The formation and storage temperatures of Clostridium botulinum spores are shown to influence their subsequent ability to germinate. Spores were formed at 10 degrees, 20 degrees, 30 degrees and 37 degrees C and following harvest were stored as aqueous suspensions at 20 degrees C (ambient temperature), 4 degrees C (refrigerated) or -20 degrees C (frozen) for periods of up to 1 month. The spores formed at 20 degrees C germinated most rapidly and to the greatest extent. When the spores were germinated immediately after harvest (fresh), there was no difference in the germinability of those spores formed at 20 degrees or 30 degrees C, whether or not they had been heat-shocked before use. However, following storage overnight or longer, differences in the relative germinabilities of the different spore samples were seen. Spores which had been stored at ambient temperature overnight germinated significantly faster and to a greater extent than did those which had been stored for up to 1 month. Similar differences were also observed between spores germinated fresh and those stored overnight, when the spores were stored refrigerated or frozen. Germinability was also influenced by the temperature of storage, since there were differences between spores formed at the same temperature but stored at different temperatures for the same period of time: for example, when spores which had been formed at 20 degrees C were germinated at 10 degrees C following a heat-shock, those which had been stored at ambient temperature germinated faster and to a greater extent than did those which had been stored refrigerated or frozen. It is concluded that there is a complex interaction between formation, storage and germination temperatures, which determines spore germinability. The fact that the changes are time-dependent and can occur in the frozen state is taken to mean that they are physico-chemical rather than metabolic. It is also significant in relation to refrigerated foods which are at risk from Cl. botulinum in that changes which occur during cool or frozen storage can enhance the germinability of spores if the temperature rises above that of chill cabinets. PMID- 9351208 TI - The contribution of moulds and yeasts to the fermentation of 'agbelima' cassava dough. AB - Agbelima, a fermented cassava meal widely consumed in Ghana, Togo and Benin, is produced by fermenting grated cassava with one of several types of traditional cassava dough inoculum. During fermentation a smooth textured sour dough is produced, the toxicity of cassava is reduced and there is a build up of volatile aroma compounds. Four types of inocula were included in the present investigation. In one type moulds were found to form a dominant part of the microbiota, the species present being Penicillium sclerotiorum, P. citrinum, P. nodulum, Geotrichum candidum and a basidiomycete. All these moulds were found to possess cellulase activity which was responsible for the hydrolysis of cassava tuber cellulose during fermentation leading to a breakdown of the coarse texture of cassava dough. The yeasts Candida krusei, C. tropicalis and Zygosaccharomyces spp. were present in high numbers in the four types of inocula including the moudly inoculum. The yeasts C. tropicalis and some strains of Zygosaccharomyces, all of which possessed cellulase activity, were also found to contribute to the modification of cassava texture during fermentation. All yeasts and moulds exhibited linamarase activity and were therefore capable of breaking down the cyanogenic glucosides present in cassava. PMID- 9351209 TI - Detection, distribution and probable fate of Escherichia coli O157 from asymptomatic cattle on a dairy farm. AB - The use of commercial anti-Escherichia coli O157-labelled magnetic beads was investigated to improve detection of E. coli O157 by immunomagnetic separation (IMS) from a range of environments on a dairy farm. Immunomagnetic separation proved effective for separation of target cells from laboratory mixtures and during stress in sterile and non-sterile pond water. The IMS procedure was possible with a range of samples (water, faeces, slurry, grass and soil). Non specific binding of non-target bacterial cells proved problematic in a number of sample types. However, indigenous E. coli O157 cells were detected from samples with a high faecal load, and only with use of IMS. Data on the probable survival and spread of the organism around the farm environment are also discussed. PMID- 9351210 TI - Effect of NaCl-tolerant lactic acid bacteria and NaCl on the fermentation characteristics and aerobic stability of silage. AB - NaCl-tolerant lactic acid bacteria (LAB) strains LC-10 (Lactobacillus casei) and LP-15 (Lact. plantarum) and NaCl were used as additives to sorghun (Sorghum bicolor). Numbers of LAB were significantly (P < 0.05) higher in all the additive treated silages than in the control silage at an early stage of ensiling. During the fermentation process, addition of NaCl or LAB effectively inhibited the growth of aerobic bacteria and clostridia, but not yeasts. All the additive treated silages had significantly (P < 0.05) lower pH, ammonia nitrogen content, dry matter loss and gas production but significantly (P < 0.05) higher lactic acid content and residual water soluble carbohydrates compared with the control silage. The improvement in silage quality was in the order: LAB > NaCl > control. Yeast counts were high in all additive-based silages and they increased during the exposure of the silages to air. As a result, these silages suffered aerobic deterioration, whereas the control silage was stable. The results confirmed that the NaCl or LAB improved fermentation quality but did not prevent aerobic deterioration of the silage. PMID- 9351211 TI - A rapid, sensitive and automated method for detection of Salmonella species in foods using AG-9600 AmpliSensor Analyzer. AB - The AG-9600 AmpliSensor Analyzer is an automated fluorescence-based system for detection of polymerase chain reaction (PCR) products. The principle of the AmpliSensor PCR assay involves amplification-mediated disruption of a fluorogenic DNA signal duplex (AmpliSensor) that is homologous to a target sequence within a 284-bp amplified fragment of the Salmonella invA gene. Since the assay is homogenous, the data can be obtained by direct measurement of fluorescence of the amplification mixture. The accumulation of the amplified product, reflected by the fluorescence index, is monitored cycle by cycle by the AG-9600 Analyzer. The detection limit of the assay was less than 2 colony-forming units (cfu) per PCR reaction using a pure culture of Salmonella typhimurium. In post-spiking experiments in which Salmonella was added to the overnight pre-enriched samples (chicken carcass rinses, ground beef, ground pork and raw milk), the detection limit of the assay was 2-6 cfu per PCR reaction. In pre-spiking experiments in which Salmonella was added to the samples prior to overnight pre-enrichment, the detection limit was less than 3 cfu per 25 g or 25 ml of food. The assay was up to 2 orders of magnitude more sensitive than detection by ethidium bromide stained agarose gel electrophoresis. To further evaluate assay performance, 54 naturally contaminated chicken carcass rinses, 65 raw milk and six ground pork samples were tested in the study. Thirty-eight Salmonella-positive samples confirmed by the Modified Semi-solid Rappaport-Vassiliadis (MSRV) culture assay were found positive using the AmpliSensor assay. Two chicken carcass rinses found positive using the assay were MSRV-negative. In addition, relative quantification using the AmpliSensor assay was linear up to 3 logs of initial target concentration in artificially contaminated food samples. PMID- 9351212 TI - Evaluation of preservative effectiveness in pharmaceutical products: the use of a wild strain of Pseudomonas cepacia. AB - A sodium benzoate-sorbic acid preservative system of a pharmaceutical product was proved effective against a wild strain of Pseudomonas cepacia, following the official method of the Italian and British Pharmacopoeias. However, this preservative system was ineffective against a challenge of Ps. cepacia wild strain cells grown in the unpreserved pharmaceutical product and on culture media different from those described by the Pharmacopoeias. The adaptive resistance of the wild strain of Ps. cepacia was not demonstrated with a laboratory strain (ATCC 25609). In contrast, p-hydroxybenzoate-based preservative systems proved to be efficient in protecting the pharmaceutical product against a challenge of wild and laboratory strains of Ps. cepacia grown in the different conditions described above. The results obtained suggest the usefulness, in the official methods for testing pharmaceutical preservatives, of using wild microbial strains isolated from the pharmaceutical environment. Metabolic adaptive responses, capable of affecting the antimicrobial sensitivity of wild micro-organisms used to challenge the preserved product, can be detected by using cells grown in the unpreserved pharmaceutical product. PMID- 9351213 TI - Use of the Malthus conductance growth analyser to determine numbers of thermophilic streptococci on stainless steel. AB - The use of the Malthus conductance growth analyser for the detection of Streptococcus bovis attached to stainless steel surface was evaluated. A comparison between the results from acridine orange epifluorescence direct counts, swab recovery viable count and conductance estimates of attached cell concentrations, based on calibrations for planktonic cells, showed that the conductance results were up to 2 log10 greater than the epifluorescence results and the swab counts. The growth rates of planktonic and attached cells were similar over 16 h using the Malthus technique. This suggests that the Malthus technique detects more attached cells of Strep. bovis than epifluorescence microscopy or swab recovery. PMID- 9351214 TI - Detection and enumeration of viable but non-culturable transconjugants of Escherichia coli during the survival of recipient cells in river water. AB - Viable but non-culturable transconjugant cells were detected by a modification of the direct viable count (DVC) method. This modification involved the addition of parental antimicrobial markers (kanamycin and streptomycin) to the elongation medium in order to promote selective elongation of the transconjugant cells. Presence of viable, other than culturable, transconjugants was demonstrated in matings with parental cells from TSB culture as well as with recipient cells from survival in river water (under illuminated and non-illuminated systems). In matings with a recipient strain from illuminated systems, culturable transconjugants were not detected after the third day of recipient cell survival. In spite of this, viable transconjugants were detected in numbers that exceeded 10(5) cells ml-1. These results clearly show that a fraction of non-culturable recipient cells is able to receive and express plasmids by conjugation processes and form viable but non-culturable transconjugant cells. PMID- 9351215 TI - Comparison between the evaluation of bacterial regrowth capability in a turbidimeter and biodegradable dissolved organic carbon bioreactor measurements in water. AB - In recent years, two different approaches to the study of biodegradable organic matter in distribution systems have been followed. The assimilable organic carbon (AOC) indicates the portion of the dissolved organic matter used by bacteria and converted to biomass, which is directly measured as total bacteria, active bacteria or colony-forming units and indirectly as ATP or increase in turbidity. In contrast, the biodegradable dissolved organic carbon (BDOC) is the portion of the dissolved organic carbon that can be mineralized by heterotrophic microorganisms, and it is measured as the difference between the inflow and the outflow of a bioreactor. In this study, at different steps in a water treatment plant, the bacterial regrowth capability was determined by the AOC method that measures the maximum growth rate by using a computerized Monitek turbidimeter. The BDOC was determined using a plug flow bioreactor. Measurements of colony forming units and total organic carbon (TOC) evolution in a turbidimeter and of colony-forming units at the inflow/outflow of the bioreactor were also performed, calculating at all sampling points the coefficient yield (Y = cfu/delta TOC) in both systems. The correlations between the results from the bioreactor and turbidimeter have been calculated; a high correlation level was observed between BDOC values and all the other parameters, except for Y calculated from bacterial suspension measured in the turbidimeter. PMID- 9351216 TI - Utilization of starch and synthesis of a combined amylase/alpha-glucosidase by the human colonic anaerobe Bacteroides ovatus. AB - Bacteroides ovatus preferentially utilized starch and pectin when grown on a mixture of polysaccharides in batch culture, indicating that these carbohydrates are important substrates for the bacterium in the human large intestine. Further studies on starch breakdown showed that continuous cultures grew on the polysaccharide when it provided the sole carbohydrate source, to yield a single hydrolytic product at low dilution rates (D = 0.04 h-1), with an estimated molecular mass of 13 kDa. In contrast, two major types of oligomeric products were formed at higher dilution rates (D = 0.44 h-1), with approximate molecular weights of 11 and 140 kDa. Analysis of cell-associated starch-degrading enzymes produced by Bact. ovatus using ion exchange chromatography and HPLC gel filtration showed that amylase and alpha-glucosidase activities eluted in the same fractions. The single peak containing amylase and alpha-glucosidase activities obtained by HPLC gel-filtration chromatography corresponded to a molecular mass of approximately 140 kDa, and activity staining of gels for alpha glucosidase activity after polyacrylamide gel electrophoresis, in the presence of sodium dodecyl sulphate, gave an estimated molecular mass of 70 kDa, indicating this enzyme to be a dimer. After renaturation, the 70 kDa band was cut from the gels and solubilized. The extract hydrolysed gelatinized starch and p-nitrophenyl alpha-D-glucopyranoside. PMID- 9351217 TI - Growth of enterobacteria on fructo-oligosaccharides. AB - Fructo-oligosaccharides (FOS) can be fermented by most species of enterobacteria present in the human intestine. Fermentation was confirmed by increased growth rates, low final pH and degradation patterns using high performance anion exchange chromatography (HPAEC). Growth rates were increased when FOS was added to the growth medium. Growth rates on all substrates differed widely between strains within the same species. HPAEC analysis showed that each strain degraded the oligosaccharides differently, but a preference for the smaller oligosaccharides was observed. No differences were observed between the two commercial preparations, the inulo-oligosaccharides and neosugars. Fermentation was rapid as could be determined by acidification tests using cell suspensions. It can be concluded that enterobacteria may play a role in overall fermentation of FOS in the colon and, in addition, due to competitive exclusion, may prevent survival of ingested pathogenic enterobacteria. PMID- 9351218 TI - Polymerase chain reaction detection and speciation of Campylobacter upsaliensis and C. helveticus in human faeces and comparison with culture techniques. AB - A polymerase chain reaction (PCR) assay based on the 16S rRNA gene and an improved DNA extraction procedure were developed for the direct detection and differentiation of Campylobacter upsaliensis and C. helveticus in seeded human faeces. The PCR assay was compared with culture detection by a membrane filter (MF) technique and on selective agar (SA) containing 8 mg l-1 cefoperazone. Both MF culture and the PCR assay detected 10(5) colony-forming units (cfu) g-1 faeces. Selective agar culture of some strains could detect as few as 10(3) cfu g 1 faeces. However, some strains were susceptible to cefoperazone and either failed to grow or were detected only with reduced sensitivity in the presence of the antibiotic. Detection by MF and SA both required 48-96 h incubation in a microaerobic atmosphere and did not specifically identify the isolate. By contrast, the PCR assay could be completed within 8 h and accurately identified the two phenotypically similar species, C. upsaliensis and C. helveticus. PMID- 9351219 TI - Subtyping of Listeria monocytogenes on the basis of plasmid profiles and arsenic and cadmium susceptibility. AB - The susceptibilities to arsenic and cadmium together with the detection of plasmid DNA were evaluated for use as epidemiological markers for the subtyping of Listeria monocytogenes. Plasmid DNA was detected in 34% of 322 apparently unrelated isolates of L. monocytogenes. The resistance to cadmium and arsenic differentiated 565 apparently unrelated cultures into four groups, the smallest being 5% of cultures resistant to both agents, and the largest (53%) being sensitive to cadmium and resistant to arsenic. The resistance patterns to these agents and the presence of plasmid DNA varied markedly between the serotypes of the cultures. The detection of plasmid DNA was strongly associated with cadmium resistance in serogroup 1/2 cultures, but not within those of serogroup 4. Arsenic resistance was not associated with plasmid DNA. All methods were sufficiently stable to be useful for epidemiology investigations. The techniques described here offer simple methods which can be easily utilized in laboratories without a specialized expertise for this bacterium. PMID- 9351220 TI - Combined effect of bacteriocin-producing lactic acid bacteria and lactoperoxidase system activation on Listeria monocytogenes in refrigerated raw milk. AB - The bactericidal activity of three bacteriocin-producing lactic acid bacteria alone and in combination with milk lactoperoxidase (LP) system activation against Listeria monocytogenes in refrigerated raw milk was studied. After 4 d at 4 degrees C, the population of L. monocytogenes in milk inoculated with bacteriocin producing Lactococcus lactis subsp. lactis ATCC 11454, L. lactis subsp. lactis ESI 515 or Enterococcus faecalis INIA 4 was reduced by 0.21-0.24 log units. Activation of the LP system did not enhance inhibition at this temperature. After 4 d at 8 degrees C, L. monocytogenes levels in the non-activated LP system milk inoculated with L. lactis subsp. lactis ATCC 11454, L. lactis subsp. lactis ESI 515 or Ent. faecalis INIA 4 were reduced by 1.87, 1.54 and 1.11 log units compared to control milk, whereas in the activated LP system milk, this reduction was 1.99, 2.10 and 1.06, respectively. The higher nisin production by L. lactis subsp. lactis ESI 515 in milk with activated LP system than in non-activated LP system milk was responsible for the more pronounced decrease of L. monocytogenes counts in the former. PMID- 9351221 TI - Increased ELISA sensitivity using a modified extraction buffer for detection of Xanthomonas campestris pv. vesicatoria in leaf tissue. AB - In vitro and in planta sensitivity of an indirect enzyme-linked immunoassay technique, using a monoclonal antibody specific for the lipopolysaccharide (LPS) of Xanthomonas campestris pv. vesicatoria, was increased 10-fold by using a new extraction buffer (gl of: KH2PO4, 2; NaHPO4, 11.5; EDTA disodium, 0.14; thimerosal, 0.02; and lysozyme, 0.2). The procedure improved sensitivity without increasing background levels. In vitro, the limit of detection was between 1 x 10(7) and 1 x 10(8) cells ml-1 with the conventional extraction buffer phosphate buffered saline (PBS) and less than 1 x 10(6) cells ml-1 when lysozyme extraction buffer was substituted for PBS. In comparing 22 X. c. vesicatoria strains, absorbance readings were increased close to three-fold with the lysozyme extraction buffer as opposed to PBS. When leaf tissue extract was spiked with the bacterium, the limit of detection was 1 x 10(7) cfu ml-1 and 1 x 10(8) cfu ml-1 with the lysozyme solution and PBS, respectively, as the extraction buffers. When using the lysozyme extraction buffer in combination with a commercial amplification system, the limit of detection was decreased to less than 1 x 10(5) cfu ml-1 in leaf tissue. The addition of the lysozyme and EDTA to the phosphate buffer resulted in release of a significant quantity of LPS and concomitant dramatic increase in sensitivity. The new procedure, termed lysozyme ELISA (L ELISA), should increase sensitivity of ELISA reactions where LPS is the reacting epitope. PMID- 9351222 TI - Determination of the influence of organic acids and nisin on shelf-life and microbiological safety aspects of fresh pork sausage. AB - The effect of replacing sulphur dioxide with organic acids and nisin to reduce the microbial counts in fresh pork sausage was examined. The potential of sodium citrate or sodium lactate, used singly or in combination with nisin, was also assessed in sausage inoculated with Staphylococcus aureus MMPR 3 and Salmonella kentucky AT 1. The results indicate that a combination of sodium lactate and nisin wa particularly effective in reducing total bacterial counts in this food product. It also appears that this combination provides an increased protection against common pathogenic contaminants of fresh pork sausage, i.e. Staph. aureus and Salmonella species. PMID- 9351223 TI - Characterization of the aggregation promoting factor from Lactobacillus gasseri, a vaginal isolate. AB - Lactobacillus gasseri 2459, isolated from the human vagina, exhibits a strong autoaggregating phenotype. Filter-sterilized spent supernatants of this strain promote aggregation of Lact. plantarum LL441 and Enterococcus faecalis EF. Aggregation was abolished upon exposure of the cells to proteases and, in the case of Ent. faecalis, to metaperiodate, which suggests the involvement of cell surface proteins and glycoproteins, respectively, in the aggregation phenotype. In accordance with this, a 75 kDa surface protein, and possibly another of approximately 94 kDa, appears in Lact. plantarum LL441 cultures incubated with Lact. gasseri culture supernatants. The diffusible aggregation promoting factor was purified from stationary phase culture supernatants and determined to be a 2 kDa hydrophilic peptide active at pH 3-4 and stable at neutral and acid pH. The activity was resistant to heat, chymotrypsin, chelating agents, triton X-100 and reducing agents, but sensitive to other proteases and SDS. PMID- 9351224 TI - Diversity among aromatic hydrocarbon-degrading bacteria and their meta-cleavage genes. AB - Sixty-one strains of bacteria capable of growth on 4-methyl benzoic acid (29 isolates) or naphthalene (32 isolates) as the sole source of carbon and energy were isolated from sediments and water samples from the River Tyne, UK. Random amplification of polymorphic DNA from genomic DNA extracted from the different strains demonstrated that 14 of the 4-methyl benzoate-degrading isolates were unique and the remainder fell into seven groups containing two or three isolates that produced identical banding patterns. Thirteen of the naphthalene-degrading isolates were unique and nine groups with two or three identical representatives encompassed all other isolates. Screening of the bacterial strains for the presence of genes homologous to xylE, nahC and bphC by polymerase chain reaction and dot blot hybridization demonstrated that most strains harboured xylE- and/or nahC-like genes and only a single isolate was found that did not harbour any of these genes. None of the isolates harboured bphC-like genes. It was concluded that, while considerable diversity existed in host strains isolated using a single simple enrichment procedure, the extradiol dioxygenase genes involved in aromatic ring cleavage, present in these strains, were conserved to a considerable degree. PMID- 9351225 TI - Immersion heat treatments for inactivation of Salmonella enteritidis with intact eggs. AB - The effects of water-bath immersion heat treatments on the inactivation of Salmonella enteritidis within intact shell eggs were evaluated. Six pooled strains of Salm. enteritidis (ca 3 x 10(8) cfu, inoculated near the centre of the yolk) were completely inactivated within 50-57.5 min at a bath temperature of 58 degrees C and within 65-75 min at 57 degrees C (an 8.4 to 8.5-D process per egg). Following the initial 24 to 35-min come-up period, semilogarithmic survivor curves obtained at 58 and 57 degrees C yielded apparent decimal reduction times (D-values) of 4.5 and 6.0 min, respectively. Haugh unit values increased during heating, while yolk index and albumen pH values were unaffected. Albumen clarity and functionality were affected by the thermal treatments; therefore, extended whip times would be required for meringue preparation using immersion-heated egg whites. Immersion-heated shell eggs could provide Salmonella-free ingredients for the preparation of a variety of minimally-cooked foods of interest to consumers and foodservice operators. PMID- 9351226 TI - The use of an automated growth analyser to measure recovery times of single heat injured Salmonella cells. AB - A new approach to the study of recovery times of single heat-injured Salmonella cells is described. It comprises the generation of a standard heat-injured culture, serial dilution of this culture to near extinction, inoculation of the serial dilutions across many microtitre plates and measurement of the subsequent recovery and growth using an automated turbidometric analyser. Lag times for individual cells were estimated from turbidity data using a model that accurately extrapolated the growth curve back to the starting inoculum level. Lag times were compared using a number of different commercially available pre-enrichment media. The most typical result was a very broad distribution of lag times at the single cell inoculum level, with many values in excess of 20 h. Even at an inoculum level 10-fold higher, lag times for some injured cells were estimated to be > 10 h. More significantly, it was found that some media recovered more injured cells than others and vice versa. Between the worst and best media there were as many as 3 log10 cycles difference in the number of cells recoverable. No trends were apparent linking choice of medium with performance. The implications of these findings, in relation to traditional and rapid methodology, are discussed. PMID- 9351227 TI - A selective medium for the rapid detection by an impedance technique of Pseudomonas spp. associated with poultry meat. AB - A new medium for detecting and enumerating Pseudomonas spp. associated with poultry meat spoilage by a rapid impedance technique was developed, after testing potential growth promoters for eight Pseudomonas strains and inhibitors against eight competing strains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977). Four basal media (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB)) and a synthetic medium were evaluated. Whitley impedance broth was the best basal medium for detecting variations in impedance in relation to Pseudomonas growth. The efficiency of WIB was improved by adding compounds which enhanced the growth of Pseudomonas on the synthetic medium. Among the incubation temperatures tested, 22 degrees C proved to be the best compromise between growth of Pseudomonas associated with poultry meat spoilage and inhibition of competitors. Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium: metronidazole, carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium). Preliminary results obtained from experiments with beef and pork meat showed that this medium could also be used without diamide and at an incubation temperature of 25 degrees C. The impedance technique using MCCCD medium was then compared with an official method which uses the medium of Mead and Adams (1977) on 106 samples of poultry neck skin. The linear regression coefficient between the two techniques was approximately r = 0.85. Impedance was able to detect 10(3) Pseudomonas g-1 within less than 19 h making it a promising technique for predicting poultry meat spoilage. PMID- 9351229 TI - Isolation and characterization of a cryptic plasmid from Erwinia citreus ATCC 31623. AB - A multiple-copy plasmid pPZG500 (3.8 kb) was isolated from a phytopathogenic bacterium Erwinia citreus ATCC 31623. This is the smallest plasmid so far isolated from the genus Erwinia. The plasmid was partially characterized by a set of restriction enzymes and the unique restriction sites were mapped for HindIII, EcoRI, EcoRV and XBaI, while three sites were found for BglII. Nineteen other enzymes did not cut pPZG500. By deletion analyses minimal regions required for replication (ori) and segregational stability (par) were localized on 1.4 kb EcoRV/BglII and 0.7 kb Bgl/II/EcoRI fragments, respectively. The erythromycin resistance marker (Emr) was cloned into pPZG500 and two plasmid derivatives, pPZG502 and pPZG503, were constructed expressing erythromycin resistance as a good selective marker for recombinant selection in Erw. citreus and Escherichia coli. The segregational stability of both constructed plasmids during 90 generations in E. coli JM109 and Erw. citreus C-4 showed that plasmid pPZG503 lacking the presumptive par region was lost from the population at a higher rate. The results of this study demonstrate that plasmid pPZG500 and derivatives are suitable prerequisites for the construction of useful cloning vector(s) in the genus Erwinia. PMID- 9351228 TI - Bacterial influence on the production of paralytic shellfish toxins by dinoflagellated algae. AB - This study investigated the role of intracellular and extracellular bacteria in the production of paralytic shellfish toxins by dinoflagellated algal cells. Three strains of the toxic dinoflagellate species, Alexandrium tamarense, were purified by external bacteria using penicillin G (Pen. G) at levels of 500 and 1000 p.p.m. Levels of toxicity of the resulting purified dinoflagellate cultures were similar to those of the original strains contaminated with external bacteria, indicating that the external bacteria had no influence on toxicity. No bacterial colony forming units (cfu) arose from disruption of algal cells derived from penicillin-treated cultures, indicating that intracellular bacteria were not responsible for the toxicity of cultures. PMID- 9351230 TI - Isolation and characterization of nisin-producing Lactococcus lactis subsp. lactis from bean-sprouts. AB - Bacterial isolates from bean-sprouts were screened for anti-Listeria monocytogenes bacteriocins using a well diffusion method. Thirty-four of 72 isolates inhibited the growth of L. monocytogenes Scott A. One, HPB 1688, which had the biggest inhibition zone against L. monocytogenes Scott A, was selected for subsequent analysis. Both ribotyping and DNA sequencing of 16S ribosomal RNA gene demonstrated that the isolate was Lactococcus lactis subsp. lactis. Polymerase chain reaction and nucleotide sequencing revealed that the genomic DNA of the bean-sprout isolates contained a nisin Z structural gene. In MRS broth, bean-sprout isolate HPB 1688 survived at 3-4.5 degrees C for at least 20 d, grew at 4 degrees C and produced anti-listerial compounds at 5 degrees C. When co cultured with L. monocytogenes in MRS broth, the isolate inhibited the growth of L. monocytogenes at 4 degrees C after 14 d and at 10 degrees C after 2 d. When co inoculated with 10(2) cells g-1 of L. monocytogenes on fresh-cut ready-to-eat Caesar salad, L. lactis subsp. lactis (10(8) cells g-1) was able to reduce the number of L. monocytogenes by 1-1.4 logs after storage for 10 d at 7 zero and 10 degrees C. A bacteriocin-producing Enterococcus faecium was also able to reduce the numbers of L. monocytogenes on Caesar salad, but did not act synergistically when co-inoculated with L. lactis subsp. lactis. PMID- 9351232 TI - A quantitative PCR-ELISA for the rapid enumeration of bacteria in refrigerated raw milk. AB - We have developed a quantitative PCR-ELISA for the rapid enumeration of bacteria in refrigerated raw milk using primers designed from conserved regions in the 16S ribosomal RNA gene (rRNA). The designed primers permitted the amplification of a 147 bp DNA fragment from a wide selection of bacteria which may grow in milk at refrigeration temperatures. Amplified PCR products generated using a digoxigenin labelled primer were heat-denatured before being quantified by an enzyme-linked immunosorbent assay (ELISA). A biotinylated probe immobilized onto streptavidin coated microplates was used to capture the digoxigenin-labelled fragments that were detected with a peroxidase anti-digoxigenin conjugate. Subsequent enzymic conversion of substrate gave distinct absorbency differences when assaying milk samples containing bacteria in the range 10(3)-10(7) cfu ml-1. The detection threshold for the PCR-ELISA assay developed in this work is 103 cfu ml-1. PMID- 9351231 TI - The influence of cell surface properties of thermophilic streptococci on attachment to stainless steel. AB - The quality of milk products is threatened by the formation of biofilms of thermophilic streptococci on the internal surfaces of plate heat exchangers used in milk processing. Although attachment to stainless steel surfaces is one of the first stages in the development of a biofilm, the mechanisms involved in attachment have not been reported. The cell surface properties of 12 strains of thermophilic streptococci were examined to determine their importance in attachment to stainless steel surfaces. Hydrophobicity, extracellular polysaccharide production and cell surface charge varied between the different strains but could not be related to numbers attaching. Treating the cells with sodium metaperiodate, lysozyme or trichloroacetic acid to disrupt cell surface polysaccharide had no effect on attachment. Treatment with trypsin or sodium dodecyl sulphate to remove cell surface proteins resulted in a 100-fold reduction in the number of bacteria attaching. This result suggests that the surface proteins of the thermophilic streptococci are important in their attachment to stainless steel. PMID- 9351233 TI - Improved preservation of the ram spermatozoan plasma membrane using betaine in the primary fixative. AB - Improved preservation of ram spermatozoa was obtained by filtration onto a Millipore filter followed by fixation in a fixative containing 23 mM betaine, which raised fixative osmolality only slightly. In samples fixed in control preparations with betaine omitted the plasma membrane had a ruffled appearance, while the betaine-containing fixative gave the plasma membrane a smooth contour which closely followed underlying structures. Betaine is known to function as an osmoprotectant in other situations, and may protect the cells from osmotic damage during the initial stages of fixation. PMID- 9351234 TI - Videodensitometric analysis of electron spectroscopic micrographs--a tool for detection of biologically relevant elements with high resolution. AB - Electron energy-loss spectroscopic imaging (ESI) yields high-resolution, element sensitive images. However, ESI suffers from difficulties in distinguishing element-specific and background contributions. New methods have therefore been introduced which use grey-level measurements in micrographic images for a more accurate detection of element distributions. A videodensitometric method allowed the detection of low phosphorus levels in axoplasmic neurofilaments of squid giant axons. Here we further verify these results by investigating the relationship of videodensitometry and electron energy-loss spectroscopy (EELS), particularly considering the peculiarities of these methods in terms of automatic background correction and representation of the results. Six biological specimens and two nonbiological specimens were examined both by EELS and by videodensitometry. In all cases comparable results were obtained. The overlapping PL2,3 and SL2,3 ionization edges could clearly be recognized individually by both methods, and controls showed that mass density variations within the specimens did not impair elemental analysis. Additional evidence supporting the detection of phosphorylation sites in squid neurofilaments was obtained in both EELS and videodensitometric measurements of neurofilament-enriched pellets and of aggregated axoplasmic particles. Thus, video-densitometry appears to be a useful tool for an improved exploration of the full imaging capabilities of energy filtering electron microscopy. PMID- 9351235 TI - Development of a standing-wave fluorescence microscope with high nodal plane flatness. AB - This article reports about the development and application of a standing-wave fluorescence microscope (SWFM) with high nodal plane flatness. As opposed to the uniform excitation field in conventional fluorescence microscopes as SWFM uses a standing-wave pattern of laser light. This pattern consists of alternating planar nodes and antinodes. By shifting it along the axis of the microscope a set of different fluorescent structures can be distinguished. Their axial separation may just be a fraction of a wavelength so that an SWFM allows distinction of structures which would appear axially unresolved in a conventional or confocal fluorescence microscope. An SWFM is most powerful when the axial extension of the specimen is comparable to the wavelength of light. Otherwise several planes are illuminated simultaneously and their separation is hardly feasible. The objective of this work was to develop a new SWFM instrument which allows standing-wave fluorescence microscopy with controlled high nodal plane flatness. Earlier SWFMs did not allow such a controlled flatness, which impeded image interpretation and processing. Another design goal was to build a compact, easy-to-use instrument to foster a more widespread use of this new technique. The instrument developed uses a green-emitting helium-neon laser as the light source, a piezoelectric movable beamsplitter to generate two mutually coherent laser beams of variable relative phase and two single-mode fibres to transmit these beams to the microscope. Each beam is passed on to the specimen by a planoconvex lens and an objective lens. The only reflective surface whose residual curvature could cause wavefront deformations is a dichroic beamsplitter. Nodal plane flatness is controlled via interference fringes by a procedure which is similar to the interferometric test of optical surfaces. The performance of the instrument was tested using dried and fluorescently labelled cardiac muscle cells of rats. The SWFM enabled the distinction of layers of stress fibres whose axial separation was just a fraction of a wavelength. Layers at such a small distance would lie completely within the depth-of-field of a conventional or confocal fluorescence microscope and could therefore not be distinguished by these two methods. To obtain further information from the SWFM images it would be advantageous to use the images as input-data to image processing algorithms such as conceived by Krishnamurthi et al. (Proc. SPIE, 2655, 1996, 18-25). To minimize specimen-caused nodal plane distortion, the specimen should be embedded in a medium of closely matched refractive index. The proper match of the refractive indices could be checked via the method presented here for the measurement of nodal plane flatness. For this purpose the fluorescent layer of latex beads would simply be replaced by the specimen. A combination of the developed SWFM with a specimen embedded in a medium of matched refractive index and further image processing would exploit the full potential of standing-wave fluorescence microscopy. PMID- 9351236 TI - Centenary of Gaule and Lewin, pioneers in cell counting methodology. AB - We commemorate the one hundredth anniversary of the publication of a pioneering paper on cell counting, by Gaule and Lewin. Their paper describes a new method for counting cells in tissue sections. First they found the mean number of cell profiles per cell by examining 50 selected cells in serial sections. Then they counted the total number of cell profiles in the ganglion, and finally divided this total profile number by the mean number of profiles per cell. They thought this method more accurate than counting cells by counting only profiles that showed the nucleolus because they noted that a cell's nucleolus sometimes appeared in more than one section and that a single cell could have more than one nucleolus. PMID- 9351237 TI - T-DNA tagging reveals a novel cDNA triggering cytokinin- and auxin-independent protoplast division. AB - Activation T-DNA tagging was used to generate four cytokinin-independent (cyi1-4) tobacco cell lines. Plants regenerated from the mutant lines displayed similar phenotypes: reduced apical dominance, poorly developed roots, delayed growth and flowering, and male and female sterility. Tissue culture experiments demonstrated that the mutations in the different lines uncouple cell proliferation from the effects of both cytokinin and auxin. No significant increase of cytokinin or auxin was found in transgenic calli in comparison with untransformed callus. The functional plant sequence tagged in one of the mutant lines, cyi1, was used to isolate an active cDNA, cyi1a, that was able to trigger cytokinin- and auxin independent protoplast division. Northern analysis shows that the transcript corresponding to cyi1a accumulates to high levels in the untransformed protoplasts shortly before the onset of cell division, and that these levels decrease when protoplasts reach maximum rates of cell division. A small putative open reading frame, starting with the first ATG in cyi1a and encoding a 22 amino acid peptide, has the same activity in tobacco protoplasts as the whole cDNA. This activity is destroyed by a frame shift mutation. Apparently cyi1a encodes a peptide which participates in the events downstream of a joint point of cytokinin and auxin action leading to cell division. PMID- 9351238 TI - Light-regulated expression of the pea plastocyanin gene is mediated by elements within the transcribed region of the gene. AB - Expression of the pea plastocyanin gene (PetE) is regulated by light in both pea and transgenic tobacco plants. However, the PetE promoter with the 5' untranslated leader region does not direct light-regulated expression of the GUS reporter gene in transgenic tobacco. This suggested that sequences downstream of the translation start of the PetE gene are required for light-regulated expression. To investigate this possibility the expression of a series of chimeric gene constructs in transgenic tobacco plants was examined to assess the contributions of the promoter, the 5' untranslated leader region, the coding region and the 3' region of the PetE gene to light-regulated expression. Both the coding region and the 5' untranslated leader region of the PetE gene were found to be required for full light regulation. Full light regulation of chimeric gene constructs containing the cauliflower mosaic virus (CaMV) 35S promoter required the deletion of CaMV 5' leader and polylinker sequences from the constructs. The presence of CaMV and polylinker sequences at the 5' end of the PetE leader masked the light regulation directed by the transcribed region of the pea PetE gene. PMID- 9351239 TI - Conserved expression of a TASSELSEED2 homolog in the tapetum of the dioecious Silene latifolia and Arabidopsis thaliana. AB - To investigate the genetics of male sex determination and stamen development in the dioecious plant Silene latifolia (white campion), male-specific transcripts were isolated from developing flowers by cDNA subtraction. One of the cDNAs identified, STA1, had high DNA and amino acid sequence homology to the male sex determining gene of Zea mays (maize), TASSELSEED2. Both genes are expressed in male and not in female flowers, However, they do not share the same expression pattern. The TASSELSEED2 gene product is expressed in the gynoecium primordia of male maize flowers where it is necessary for pistil abortion. STA1 is not expressed in the gynoecium primordia of male white campion and therefore its gene product cannot perform the same function in sex determination that TASSELSEED2 performs in maize. STA1 is expressed in tapetal cells of white campion male flowers and of white campion hermaphroditic mutants. A homologous gene is also expressed in the tapetum of hermaphroditic Silene species. Tapetal expression of a homologous gene (named ATA1) was also found in Arabidopsis thaliana. The similarity in primary sequence and expression pattern of STA1 and ATA1 indicate that these genes have a conserved role in tapetum development. PMID- 9351240 TI - Identification of three genetic loci controlling leaf senescence in Arabidopsis thaliana. AB - Four mutants that show the delayed leaf senescence phenotype were isolated from Arabidopsis thaliana. Genetic analyses revealed that they are all monogenic recessive mutations and fall into three complementation groups, identifying three genetic loci controlling leaf senescence in Arabidopsis. Mutations in these loci cause delay in all senescence parameters examined, including chlorophyll content, photochemical efficiency of photosystem II, relative amount of the large subunit of Rubisco, and RNase and peroxidase activity. Delay of the senescence symptoms was observed during both age-dependent in planta senescence and dark-induced artificial senescence in all of the mutant plants. The results indicate that the three genes defined by the mutations are key genetic elements controlling functional leaf senescence and provide decisive genetic evidence that leaf senescence is a genetically programmed phenomenon controlled by several monogenic loci in Arabidopsis. The results further suggest that the three genes function at a common step of age-dependent and dark-induced senescence processes. It is further shown that one of the mutations is allelic to ein2-1, an ethylene insensitive mutation, confirming the role of ethylene signal transduction pathway in leaf senescence of Arabidopsis. PMID- 9351241 TI - Movement and subcellular localization of a tobamovirus in Arabidopsis. AB - Tobamoviruses represent a well-characterized system used to examine viral infection, whereas Arabidopsis is a choice plant for most genetic experiments. It would be useful to combine both approaches into one experimental system for virus plant interaction. Most tobamoviruses, however, are not pathogenic in Arabidopsis. Here, we describe infection of Arabidopsis by a recently discovered crucifer-infecting turnip vein clearing tobamovirus (TVCV). Using this system, we determined patterns and kinetics of viral local and systemic movement within Arabidopsis plants. Localization studies showed that the virus infects both vegetative and reproductive plant tissues. However, there may be a transport barrier between the seed coat and the embryo which virions cannot cross, preventing seed transmission of TVCV. The ability to move both locally and systemically in Arabidopsis, causing mild and fast-developing symptoms but allowing survival and fertility of the infected plants, distinguish TVCV infection of Arabidopsis as a model system to study virus-plant interaction. PMID- 9351243 TI - Sites of rDNA transcription are widely dispersed through the nucleolus in Pisum sativum and can comprise single genes. AB - Incorporation by RNA polymerases of BrUTP into both plant root tissue and isolated plant nuclei as a method for localization of the sites of transcription has been used. In this paper pea root tissue was used, and under the conditions employed, nearly all the incorporation occurs in the nucleolus, and thus must be catalysed by RNA polymerase I. Immunofluorescence and confocal microscopy shows that incorporation occurs in a pattern consisting of many small foci distributed widely through the dense fibrillar component of the nucleoli. Immunogold labelling using silver-enhanced Nanogold probe at the electron microscopic level confirms the sites of transcription as small foci approximately 200 nm in diameter. Simultaneous fluorescence in situ hybridization with a probe to the external transcribed spacer (ETS) region of the pre-rRNA shows that the structures revealed by this probe and the BrUTP immunofluorescence labelling are very similar. A probe to the transcribed portion of the rDNA (18S) also shows a good correlation to the sites of BrUTP incorporation within the nucleolus. On the other hand a probe to the non-transcribed intergenic spacer region (NTS) shows very little coincidence with the sites of BrUTP incorporation, and double fluorescence in situ labelling with both 18S and NTS probes confirms this difference in localization. These results suggest that most BrUTP foci correspond to single transcribed genes. PMID- 9351242 TI - Differential expression of two P5CS genes controlling proline accumulation during salt-stress requires ABA and is regulated by ABA1, ABI1 and AXR2 in Arabidopsis. AB - Proline is a common compatible osmolyte in higher plants. Proline accumulation in response to water stress and salinity is preceded by a rapid increase of the mRNA level of delta 1-pyrroline-5-carboxylate synthase (P5CS) controlling the rate limiting step of glutamate-derived proline biosynthesis. P5CS is encoded by two differentially regulated genes in Arabidopsis. Gene AtP5CS1 mapped to chromosome 2-78.5 is expressed in most plant organs, but silent in dividing cells. Gene AtP5CS2 located close to marker m457 on chromosome 3-101.3 contributes 20-40% of total P5CS mRNA in plant tissues, but is solely responsible for the synthesis of abundant P5CS mRNA in rapidly dividing cell cultures. Accumulation of AtP5CS transcripts is regulated in a tissue specific manner and inducible by drought, salinity, ABA, and to a lesser extent by auxin. Induction of AtP5CS1 mRNA accumulation in salt-treated seedlings involves an immediate early transcriptional response regulated by ABA signalling that is not inhibited by cycloheximide, but abolished by the deficiency of ABA biosynthesis in the aba1 Arabidopsis mutant. However, inhibition of protein synthesis by cycloheximide prevents the induction of AtP5CS2 mRNA accumulation, and blocks further increase of AtP5CS1 mRNA levels during the second, slow phase of salt-induction. Mutations abi1 and axr2, affecting ABA-perception in Arabidopsis, reduce the accumulation of both AtP5CS mRNAs during salt-stress, whereas ABA-signalling functions defined by the abi2 and abi3 mutations have no effect on salt-induction of the AtP5CS genes. PMID- 9351244 TI - PIS1, a negative regulator of the action of auxin transport inhibitors in Arabidopsis thaliana. AB - In order to clarify the mechanism underlying the polar auxin transport system, the pis1 mutant in Arabidopsis thaliana that is hypersensitive to N-1 naphthylphthalamic acid (NPA), an auxin transport inhibitor was isolated and characterized. Whereas the pis1 mutant is normally sensitive to phytohormones, auxins, cytokinin and ethylene precursor, this mutant is hypersensitive to NPA over the broad spectrum of its effects such as growth of seedlings, root elongation, root gravitropism, root phototropism and root curling. This result indicates that the pis1 mutant is specifically affected in the polar auxin transport system. This result also defines a genetic factor controlling both gravitropism and phototropism, and strongly indicates the involvement of auxin transport during both tropic responses. NPA, 2,3,5-triiodobenzoic acid (TIBA) and 9-hydroxyfluorene-9-carboxylic acid (HFCA) represent different classes of auxin transport inhibitors. The pis1 mutation conferred hypersensitivity to both NPA and TIBA but not to HFCA. These results show the genetic separation of the actions of NPA/TIBA and of HFCA. The PIS1 gene product might be specifically involved in the response pathway of NPA/TIBA, leading to interference with auxin efflux carriers, and might act as a negative regulator of the action of NPA/TIBA. PMID- 9351245 TI - Development of Arabidopsis thaliana leaves at low temperatures releases the suppression of photosynthesis and photosynthetic gene expression despite the accumulation of soluble carbohydrates. AB - Arabidopsis thaliana plants were grown at 23 degrees C and changes in carbohydrate metabolism, photosynthesis and photosynthetic gene expression were studied after the plants were shifted to 5 degrees C. The responses of leaves shifted to 5 degrees C after development at 23 degrees C are compared to leaves that developed at 5 degrees C. Shifting warm developed leaves to 5 degrees C lead to a severe suppression of photosynthesis that correlated with a rapid and sustained accumulation of hexose phosphates and soluble sugars. Associated with the suppression of photosynthesis and the accumulation of soluble sugars was a reduction in the amount of transcript for genes encoding photosynthetic proteins (cab and rbcS). In contrast, leaves that developed at 5 degrees C showed an increase in photosynthesis and control levels of photosynthetic gene expression. This recovery occurred even though leaves that developed at 5 degrees C maintained large pools of soluble sugars. Leaves that developed at 5 degrees C also showed a strong upregulation of the cytosolic pathway for soluble sugar synthesis but not of the chloroplastic pathway for starch synthesis. This was shown at the level of both enzyme activity and the amount of transcript. Thus, development of Arabidopsis leaves at 5 degrees C resulted in metabolic changes that enabled them to produce and accumulate large soluble sugar pools without any associated suppression of photosynthesis or photosynthetic gene expression. These changes were also associated with enhanced freezing tolerance. We suggest that this reprogramming of carbohydrate metabolism associated with development at low temperature is essential to the development of full freezing tolerance and for winter survival of over-wintering herbaceous annuals. PMID- 9351246 TI - The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. AB - The Arabidopsis thaliana MALE STERILITY 2 (MS2) gene product is involved in male gametogenesis. The first abnormalities in pollen development of ms2 mutants are seen at the stage in microsporogenesis when microspores are released from tetrads. Expression of the MS2 gene is observed in tapetum of wild-type flowers at, and shortly after, the release of microspores from tetrads. The MS2 promoter controls GUS expression at a comparable stage in the tapetum of transgenic tobacco containing an MS2 promoter-GUS fusion. The occasional pollen grains produced by mutant ms2 plants have very thin pollen walls. They are also sensitive to acetolysis treatment, which is a test for the presence of an exine layer. The MS2 gene product shows sequence similarity to a jojoba protein that converts wax fatty acids to fatty alcohols. A possible function of the MS2 protein as a fatty acyl reductase in the formation of pollen wall substances is discussed. PMID- 9351247 TI - Light-dependent regulation of carotenoid biosynthesis occurs at the level of phytoene synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana seedlings. AB - In chloroplasts, carotenoids are essential pigments involved in photosynthesis. During-photomorphogenesis, a coordinated increase in the amounts of chlorophylls and carotenoids, in conjugation with other components, leads to the formation of a functional photosynthetic apparatus. To investigate the regulation of carotenoid biosynthesis during this process at the molecular level, GGPS, PSY and PDS cDNAs have been cloned from white mustard (Sinapis alba L). GGPS encodes a key enzyme in plastid isoprenoid metabolism, while the products of PSY and PDS catalyse the subsequent steps in carotenoid biosynthesis. Due to the low mRNA levels of the genes involved, the use of a RT-PCR protocol was necessary to measure gene expression during photomorphogenesis. With light, there is an up regulation of PSY expression, the first gene within the carotenoid biosynthetic pathway, while PDS and GGPS expression levels remain constant. Treatment with different light qualities reveals a phytochrome-mediated regulation of PSY expression in developing white mustard seedlings. To obtain more detailed information on the light-regulation, Arabidopsis thaliana wild-type and phytochrome mutants were utilized. Continuous far-red and red light both increase the expression of PSY in wild-type seedlings, demonstrating that both light labile and light-stable phytochromes are involved in PSY regulation. The response to far-red light is completely abolished in the phyA mutant, showing that PHYA mediates the increase in PSY transcript levels under these light conditions. In the phyB mutant, the red light response is normal, indicating that PSY expression is not controlled by PHYB but by other light-stable phytochromes. Measurement of chlorophylls and carotenoids under the same light regimes shows that the up regulation of PSY expression does not necessarily result in an increase of the carotenoid content. Only those light conditions which allow chlorophyll biosynthesis lead to a significant increase of the carotenoid content. Therefore, it is proposed that up-regulation of PSY mRNA levels leads to an increased capacity for the formation of carotenoids. However, this only takes place under light conditions leading to protochlorophyllide photoconversion. PMID- 9351248 TI - Developmental and auxin-induced expression of the Arabidopsis prha homeobox gene. AB - A single-copy Arabidopsis homeobox gene, prha, which encodes a homeodomain protein with a molecular weight of 90,500 has been characterized. The position of the gene was mapped to the distal part of chromosome 4. Expression of the gene differs in various vegetative and floral plant tissues and is positively influenced by the phytohormone auxin. In Arabidopsis plants, a complex pattern of prha promoter-driven GUS expression is observed, often associated with regions of developing vascular tissue. Exogenously applied auxin strongly increased endogenous prha transcript levels. In addition, the prha promoter is highly responsive to the synthetic auxin, naphthalene acetic acid, in transient assays using tobacco protoplasts. The PRHA protein has the capacity to bind to TAATTG core sequence elements but requires additional adjacent bases for high-affinity binding. These findings are discussed in relation to studies of other plant homeobox genes as well as possible in vivo target genes for PRHA. PMID- 9351249 TI - Overexpression of light-dependent PORA or PORB in plants depleted of endogenous POR by far-red light enhances seedling survival in white light and protects against photooxidative damage. AB - The structurally related light-dependent protochlorophyllide (Pchlide) oxidoreductases PORA and PORB mediate the only light-requiring step in chlorophyll (Chl) biosynthesis in higher plants. Correlative evidence suggests that some in vivo functions of PORA and PORB may be unique, including a postulated photoprotective role for PORA. For example, wild-type Arabidopsis thaliana seedlings grown in non-photooxidative far-red light (cFR) resemble those grown in white light (WL), but they are yellow and do not green normally thereafter in WL. This defect is accompanied by the absence of detectable PORA and reduced levels of PORB expression. Here, direct evidence is provided that the presence of POR, either as PORA or PORB, can confer photoprotection in plants. In contrast to the wild-type, the plastids of transgenic PORA- or PORB overexpressing Arabidopsis seedlings grown in cFR possess extensive prolamellar bodies. Upon a subsequent shift to WL, POR-overexpressing seedlings develop thylakoid membranes, accumulate large amounts of Chl and are viable at fluence rates lethal to the wild-type. Intriguingly, the plastid membrane architectures of greening transgenic seedlings seem to depend on whether PORA or PORB has been overproduced. POR-overexpressing seedlings shifted from cFR to WL of fluence rates from 20 to 500 muE m-2 sec-1 accumulate substantially higher amounts of Chl than does the wild-type. Furthermore, the WL fluence rate that permits maximal Chl accumulation increases from 8 muE m-2 sec-1 in the wild-type to 125 muE m-2 sec-1 in transgenic seedlings. POR overexpression during growth in cFR also correlates with a fourfold decrease in the steady-state content of Pchlide, a potentially lethal photosensitizer. PMID- 9351250 TI - The VLF loci, polymorphic between ecotypes Landsberg erecta and Columbia, dissect two branches of phytochrome A signal transduction that correspond to very-low fluence and high-irradiance responses. AB - Phytochromes play a key role in the perception of light signals by plants. In this study, the three classical phytochrome action modes, i.e. very-low-fluence responses (VLFR), low-fluence responses (LFR) and high-irradiance responses (HIR), were genetically dissected using phyA and phyB mutants of Arabidopsis thaliana (respectively lacking phytochrome A or phytochrome B) and a polymorphism between ecotypes Landsberg erecta and Columbia. Seed germination and potentiation of greening, hypocotyl growth inhibition and cotyledon unfolding in etiolated seedlings of the ecotype Landsberg erecta showed biphasic responses to the calculated proportion of active phytochrome established by one light pulse or repeated light pulses. The first phase, i.e. the VLFR, was absent in the phyA mutant, normal in the phyB mutant (both in the Landsberg erecta background) and severely deficient in Columbia. The second phase, i.e. the LFR, was present in the phyA mutant, deficient in the phyB mutant and normal in Columbia. Under continuous far-red light, HIR of etiolated seedlings were absent in phyA and normal in phyB and Columbia. The segregation of VLFR in recombinant inbred lines derived from a cross between Landsberg erecta and Columbia was analysed by MAPMAKER/QTL. Two quantitative trait loci, one on chromosome 2 (VLF1) and another on chromosome 5 (VLF2), were identified as responsible for the polymorphism. Phytochrome A is proposed to initiate two transduction pathways, VLFR and HIR, involving different cells and/or different molecular steps. This is the first application of the analysis of quantitative trait loci polymorphic between ecotypes to dissect transduction chains of environmental signals. PMID- 9351251 TI - Enhanced tolerance to bacterial pathogens caused by the transgenic expression of barley lipid transfer protein LTP2. AB - Purified lipid transfer protein LTP2 from barley applied on tobacco leaves eliminated symptoms caused by infiltration of Pseudomonas syringae pv. tabaci 153. Growth of the pathogen in leaves of transgenic tobacco plants was retarded when compared with non-transformed controls. The percentage of inoculation points that showed necrotic lesions was greatly reduced in transgenic tobacco (17-38% versus 78%) and the average size of these lesions was 61-81% that of control. The average total lesion area (necrosis and chlorosis) in the transgenic plants was also reduced (38% of control). Arabidopsis thaliana transgenic plants inoculated with P. syringae pv. tomato DC3000 also had lower percentages of necrotic lesions (22-38% versus 76%), a reduced average area for each lesion (53-67% of control), and a smaller total lesion area per inoculation (43% of control). These results further support the assignment of a defense role for LTPs and highlight their biotechnological potential. PMID- 9351252 TI - Cloning and characterization of LRG5, a gene involved in blue light signaling in Chlamydomonas gametogenesis. AB - The Chlamydomonas reinhardtii mutant lrg2 exhibits a partial defect in the blue light activated signal transduction chain that controls gametogenesis. By genomic complementation, a cosmid clone that corrects the lrg2 phenotype was isolated. The smallest fragment of this cosmid that gave complementation was used in RFLP analysis, which revealed that the cloned gene was not linked to the LRG2 locus. The sequence of the genomic clone and a cDNA of this gene, which we named LRG5, was determined. Although no significant homology with sequences in the databases was found, the nuclear localization signal and the presence of negatively and positively charged domains suggests a function for the LRG5 protein in the regulation of transcription. The correction of the lrg2 phenotype is probably caused by overexpression of the LRG5 gene in the transformants, as these expressed a level of LRG5 approximately twofold higher than wild-type or mutant strains. Southern blot analysis provided evidence for the existence of homologous sequences in other green algae as well as in higher plants, suggesting a conserved function for the LRG5 protein. PMID- 9351253 TI - A negative selection scheme based on the expression of cytosine deaminase in plastids. AB - The enzyme cytosine deaminase (CD) encoded by codA catalyzes deamination of cytosine to uracil. CD is present in prokaryotes and in many eukaryotic micro organisms, but is absent in higher plants. 5-fluorocytosine (5FC) is metabolized in CD-expressing cells, causing cellular death. A chimeric codA has been introduced into the tobacco plastid genome and 5FC was used to select against tissue culture cells and seedlings expressing CD. This negative selection scheme will be useful in identifying nuclear genes which control plastid gene expression in higher plants. PMID- 9351254 TI - Microdissection and amplification of coding sequences from a chromosome fragment restoring male fertility in alloplasmic male-sterile tobacco. AB - An alloplasmic male-sterile line of tobacco, containing the nucleus of Nicotiana tabacum and the cytoplasm of Nicotiana repanda, is restored to fertility by introgression of an alien chromosome fragment obtained from the cytoplasm donor. To isolate the restorer gene(s), the alien chromosome fragment was microdissected from metaphase plates of the restored line. The microdissected chromosomes represented only 0.1 pg of DNA, which was amplified using a degenerate oligonucleotide-primed PCR method (DOP-PCR), from which a chromosome fragment specific library was created. Compared with previous strategies used for microcloning, a modified and improved method was developed by the subsequent isolation of expressed sequences. The library was screened with cDNA probes synthesized by reverse transcription and DOP-PCR amplification (RT/DOP-PCR), of total RNAs isolated from early developing restored and male-sterile flower buds. By this strategy, transcribed DNA sequences specific for the restored line were cloned. PMID- 9351255 TI - Construction of an approximately 2 Mb contig in the region around 80 cM of Arabidopsis thaliana chromosome 2. AB - A method for construction of bacterial artificial chromosome (BAC) contigs from a yeast artificial chromosome (YAC) physical map is described. An approximately 2 Mb contig, consisting of two large BAC contigs linked by a small YAC, has been assembled in the region around 80 cM of Arabidopsis thaliana chromosome 2. Clones from this contig will facilitate gene isolation in the region and can be used directly as substrates for DNA sequencing. PMID- 9351256 TI - Isolation and characterization of a transposon mutant of Shewanella putrefaciens MR-1 deficient in fumarate reductase. AB - A transposon mutant, designated CMTn-3, of Shewanella putrefaciens MR-1 that was deficient in fumarate reduction was isolated and characterized. In contrast to the wild-type, CMTn-3 could not grow anaerobically with fumarate as the electron acceptor, and it lacked benzyl viologen-linked fumarate reductase activity. Consistent with this, CMTn-3 lacked a 65 kDa c-type cytochrome, which is the same size as the fumarate reductase enzyme. CMTn-3 retained the wild-type ability to use nitrate, iron(III), manganese(IV) and trimethylamine N-oxide (TMAO) as terminal electron acceptors. The results indicate that the loss of the fumarate reductase enzyme does not affect other anaerobic electron transport systems in this bacterium. PMID- 9351257 TI - Nisin production by Lactococcus lactis using two-phase batch culture. AB - Nisin production by Lactococcus lactis subsp. lactis C2SmPrt-(TnNip) was investigated using two-phase batch culture. A solvent (phenyl-methyl silicone oil) was introduced in addition to the aqueous phase. The partition coefficient of this solvent in aqueous nisin varied as the cultivation medium changed, with a minimum value being 1.04. The two-phase batch culture supported a 21% increase in growth and a 24% increase in the level of nisin produced compared to the single phase batch culture. PMID- 9351258 TI - Extraction of lithium from spodumene by bioleaching. AB - The recovery of lithium from spodumene (6.9% Li2O) by bioleaching was investigated. This process was carried out using heterotrophic micro-organisms previously isolated from the mineral. Penicillium purpurogenum, Aspergillus niger and Rhodotorula rubra were assayed separately. Two different media were used for bioleaching; one of them (M2 medium) was highly limited in Mg2+, Fe2+ and K+. The assays were carried out in 500 ml Erlenmeyer flasks with 1 g of ground mineral at 50-80 mesh and 150 ml of leaching medium. Lithium extracted and accumulated in biomass during 30 d of bioleaching with P. purpurogenum was 6.35 mg % dry weight (d.w.) in M1 medium and 10.8 mg % d.w. in M2 medium, while in the leach liquor, Li concentration was 1.06 ppm (M1 medium) and 1.26 ppm (M2 medium). Results of leaching on day 30 with R. rubra were 5.87 mg % d.w. and 16.7 mg % d.w. of lithium accumulated in biomass in M1 medium and M2 medium, respectively. In leach liquor, lithium was 0.5 ppm (M1 medium) and 1.53 ppm (M2 medium). Aspergillus niger was able to accumulate 1.60 mg % d.w. (M1 medium) and 5.1 mg % d.w. of lithium (M2 medium) in biomass. Lithium in leach liquor was 0.37 ppm (M1 medium) and 0.75 ppm (M2 medium). Chemical analysis of the leach liquor showed gluconic and citric acids. It was possible to detect capsular exopolymers in the yeast. These metabolic products seem to be related to leaching but a more important factor for enhancing this process may be microbial adaptation to a low nutrient environment. PMID- 9351260 TI - Alveolar macrophage reaction to Candida species. AB - Candida species are increasingly important fungal pathogens. The reaction of rat alveolar macrophages (AM) to Candida albicans was compared with that to C. glabrata and C. krusei. Phagocytosis of C. glabrata was similar to that of C. albicans, but significantly slower for C. krusei due to reduced attachment. After opsonization, attachment of C. albicans and C. krusei to AM was significantly increased and there was no significant difference between the two species. The oxidative metabolism of AM with candida species was two to three times higher than that of the resting AM both during and 24 h after the phagocytosis. All three species showed a considerable fraction (5-10%) of phagolysosomes with pH > or = 6.5 after 3 h and a smaller percentage (1%) after 24 h. PMID- 9351259 TI - Growth inhibition of Helicobacter pylori by a polyamine synthesis inhibitor, methylglyoxal bis(cyclopentylamidinohydrazone). AB - The anti-proliferative effect of methylglyoxal bis(cyclopentylamidino-hydrazone) (MGBCP), a multi-enzyme inhibitor of polyamine biosynthesis, on the growth of Helicobacter pylori was investigated. MGBCP inhibited the cell growth of H. pylori in a dose-dependent manner. The inhibition was partially reversed by the addition of spermidine. Synthesis of macromolecules, DNA, RNA and protein, was inhibited in the spermidine-depleted H. pylori cells. These findings suggest that MGBCP exhibits an anti-proliferative effect on H. pylori by suppression of macromolecule synthesis. PMID- 9351261 TI - Detection of Clostridium botulinum types A, B, E and F in foods by PCR and DNA probe. AB - A PCR procedure was developed for the detection of Clostridium botulinum in foods. PCR products were detected in agarose gels and by Southern hybridization. The sensitivity of PCR was tested in broth cultures and in canned asparagus, dry cured ham and honey. The sensitivity of the method in broth was high (2.1-8.1 cfu ml-1) for types A and B, but rather low (10(4) cfu ml-1) for types E and F. However, after enrichment at 37 degrees C for 18 h, it was possible to detect Cl. botulinum types A, B, E and F in food samples at initial levels of about 1 cfu 10 g-1 of food. This PCR detection protocol provides a sensitive and relatively rapid technique for the routine detection of Cl. botulinum in foods. PMID- 9351262 TI - The effect of yellow affinity substance on cellulases of Ruminococcus flavefaciens. AB - Cellulolytic cultures of Ruminococcus flavefaciens produced a yellow affinity substance (YAS) with a strong affinity to microcrystalline cellulose (MC). YAS was bound to MC in the range of pH from 5 to 8 and at temperatures from 10 degrees C to 60 degrees C. The positive effect of YAS on adsorption of ruminococcal cellulases was demonstrated by comparing the adsorption behaviour of endoglucanases and cellobiohydrolases onto MC and YAS-treated MC. HPLC chromatography proved the presence of two yellow compounds with affinity to cellulose as well as to ruminococcal cellulases. Both YAS compounds were sensitive to oxidation. The observed YAS properties showed a close relation to YS of Clostridium thermocellum. PMID- 9351263 TI - A Penicillium freii gene that is highly similar to the beta-ketoacyl synthase domain of polyketide synthase genes from other fungi. AB - A Penicillium freii DNA fragment with similarity to the beta-keto-acyl synthase motif of the P. griseofulvum 6-methylsalicylic acid synthase encoding gene (MSAS) was identified by screening a cosmid library using a part of MSAS as the probe. Two exons of 93 and 849 bp, encoding a predicted polypeptide of 314 amino acids, and a molecular mass of 33.4 kDa, were identified (PfKS). PfKS was transcribed as a 1.6 kbp messenger. A region corresponding to the MSAS gene encoding essential enzyme activities for the assembly of a polyketide chain was absent in PfKS. Gene disruption experiments in P. freii with a truncated version of PfKS did not result in the detection of homologous integration events. PMID- 9351264 TI - Novel selective and non-selective optical detection of microorganisms. AB - A new instrument, capable of detecting metabolic changes due to microbiological activity, is described. Optical changes in growth media are monitored in a semi fluid zone that separates the liquid medium containing the sample. Data demonstrate that common media can be utilized in conjunction with this rapid automated technology. Nutrient broth with the pH dye indicator. bromocresol purple was suitable for total counts. Selective media containing dyes were utilized to assess the presence or absence of specific groups of organisms. Biochemical reactions, such as lysine decarboxylase activity, were identified by the unique generated patterns, and specific enzymatic cleavage reactions with chromogenic substrates, such as 5-bromo-4 chloro-3 indolyl-beta-D-glucuronic acid (X-GLUC), were monitored. PMID- 9351265 TI - Effect of medium composition, agitation and the presence of EDTA on the antimicrobial activity of cryptolepine. AB - The antimicrobial activity of cryptolepine is influenced by the type of medium employed, agitation and the presence of non-inhibitory concentrations of EDTA. The use of Mueller-Hinton broth (MHB), iso-sensitest broth and tryptone soya broth (TSB) produced lower minimum inhibitory concentrations (MICs) for some of the test organisms compared with nutrient broth or yeast dextrose broth (YDB). For example, a fourfold drop in MIC was recorded for Saccharomyces cerevisiae in MHB compared with the same organism tested in YDB. Agitation of the broths during incubation nearly always produced lower MICs for the bacteria, an eightfold decrease in MIC being recorded for Escherichia coli cultured in nutrient broth with agitation compared with a statically maintained culture. A non-inhibitory concentration (10(-3) mol l-1) of disodium EDTA enhanced the antimicrobial activity of cryptolepine. Against E. coli NCTC 11,560, an eightfold decrease in MIC and minimum bactericidal concentration (MBC) was recorded when tested in the presence of EDTA. PMID- 9351266 TI - Isolation and characterization of proteinase- and aminopeptidase-deficient mutants of Lactobacillus casei subsp. casei IFPL 731. AB - Proteinase-deficient (Prt-) and aminopeptidase-deficient (Amp-) variants of Lactobacillus casei subsp. casei IFPL 731 were isolated and characterized. The Prt- mutant was isolated from strains that developed poorly on glucose milk agar. The Amp- mutant was isolated on the basis of its inability to hydrolyse L-leucine beta-naphtylamide. The Prt- variant developed poorly, while in milk the Amp- variant grew at about the same rate as the parental strain. The characterization of aminopeptidase activity in more detail showed that at least two enzymes are involved The results of the present study suggest that the proteolytic system of Lactobacillus casei is subjected to a regulatory system. PMID- 9351267 TI - Partial deletion of transposon Tn4560 integrated into the genome of Streptomyces tendae. AB - Polymerase chain reaction (PCR), Southern hybridization and DNA sequencing experiments were done to determine whether all of Tn4560, a Streptomyces transposon, integrated into the genomes of three nikkomycin non-producing mutants. A deletion of 279 bases occurred at one end of Tn4560 while present in the genome of one of the mutants. PMID- 9351268 TI - Identification and characterization of homofermentative mesophilic Lactobacillus strains isolated from artisan starter-free cheeses. AB - Fifty-six strains of mesophilic lactobacilli from hand-made cheeses made without starters have been isolated, identified and characterized. Of these, 21 strains were classified as Lactobacillus plantarum, 18 as Lact. casei subsp. pseudoplantarum, 10 as Lact. curvatus, five as Lact. casei subsp. casei, and two remained unidentified. The numerical classification of these strains, based on 80 different physiological and morphological characteristics, correlated well with the phenotypic classification. Most of the technologically important traits have been examined in these strains, which will allow the selection of some of them to be tested as adjunct cultures in the manufacture of dairy products. PMID- 9351269 TI - The determination of the partial 18 S ribosomal DNA sequences of Cordyceps species. AB - Cordyceps species, which are used in Chinese traditional medicines, are fungal parasites of insects. In this study the partial nucleotide sequences of 18 S ribosomal DNA from four Cordyceps species were determined and compared with the sequences of published ascomycetes. The sequence data support the concept that Cordyceps species belong to the pyrenomycetes. Based on sequence data the phylogenetic tree was constructed using the neighbor-joining (NJ) method. Diversity in the phylogenetic tree was found for Cordyceps species. A new classification of Cordyceps species can be constructed based on the phylogenetic information obtained from such rDNA sequences. PMID- 9351270 TI - Degradation of quillaja saponins by mixed culture of rumen microbes. AB - Quillaja saponin (QS) was incubated at 39 degrees C in an in vitro medium containing rumen liquor from a cow fed a roughage diet. No degradation of QS was observed up to 6 h of fermentation. Incubation for 9, 12 and 24 h decreased the content of QS by 16%, 45% and 100%. The content of QS did not decrease when incubated for 24 h in the medium containing autoclaved rumen liquor, suggesting that rumen microbes have enzyme(s) capable fo degrading QS. The fate of QS will help gain a better understanding of mechanisms of action of QS on rumen fermentation, and its beneficial effects mediated by binding to ammonia. PMID- 9351271 TI - Use of antibody-coated cellulose sponges for enhanced isolation of salmonella. AB - One thousand, four hundred and fifty-one naturally contaminated samples from pig, poultry and cattle farms, poultry hatcheries and animal feed mills were examined in a trial in which transfer of small portions of cellulose sponge coated with salmonella somatic polyvalent antiserum was compared with transfer of standard liquid inocula from pre-enrichment to selective enrichment culture. Salmonella was found in 281 (19.4%) of the samples using the standard method, compared with 385 (26.5%) using the sponge method. It was therefore concluded that antibody coated cellulose sponges could be a simple means of increasing the recovery of salmonellas from pre-enrichment broths and thereby enhancing the test sensitivity. PMID- 9351273 TI - Yeast exoglycoproteins produced under NaCl-stress conditions as efficient cryoprotective agents. AB - Six extracellular yeast glycoproteins were prepared from three yeast species in osmotic equilibrium and unequilibrium environments and used as non-penetrating cryoadditives. The glycoprotein secreted by the strain Dipodascus australiensis into the growth medium containing NaCl (8% w/v) was found to be the most effective cryoadditive. It was possible to use this glycoprotein alone (without penetrating agent DMSO) for the cryoprotection of the yeasts studied. PMID- 9351274 TI - Significance of temperature and preincubation temperature on survival of Listeria monocytogenes at pH 4.8. AB - Listeria monocytogenes is a food-borne pathogenic bacterium that can be found in soft cheese. At the beginning of cheese ripening, the pH is about 4.85-4.90. The aim of this work was to study the influence of temperature, preincubation temperature (temperature at which the inoculum was cultivated) and initial bacterial concentration on the survival of L. monocytogenes (strain Scott A) at pH 4.8. It was demonstrated in an earlier study that these factors did influence growth kinetics. Survival studies of L. monocytogenes were done in a laboratory broth simulating cheese composition. Four test temperatures (2, 6, 10 and 14 degrees C) and two preincubation temperatures were studied (30 degrees C or the test temperature). Listeria monocytogenes (strain Scott A) was unable to grow at pH 4.8 under all conditions tested. The time for 10% survival was about 11 and 2 d, at 2 degrees C with preincubation at 2 degrees C and 30 degrees C, respectively; 9 d at 6 degrees C with preincubation at 6 degrees C; 4 d at 6 degrees C with preincubation at 30 degrees C; and 1 d at 14 degrees C with preincubation at 14 degrees C or at 30 degrees C. The results show that survival of L. monocytogenes (strain Scott A) at pH 4.8 is not dependent on initial bacterial concentration but on both the test and preincubation temperatures. PMID- 9351272 TI - Glucose respiration and fermentation in Zygosaccharomyces bailii and Saccharomyces cerevisiae express different sensitivity patterns to ethanol and acetic acid. AB - In the yeast Zygosaccharomyces bailii ISA 1307, respiration and fermentation of glucose were exponentially inhibited by ethanol, both processes displaying similar sensitivity to the alcohol. Moreover, the degree of inhibition on fermentation was of the same magnitude as that reported for Saccharomyces cerevisiae. Acetic acid also inhibited these two metabolic processes in Z. bailii, with the kinetics of inhibition again being exponential. However, inhibition of fermentation was much less pronounced than in S. cerevisiae. The values estimated with Z. bailii for the minimum inhibitory concentration of acetic acid ranged from 100 to 240 mmol 1(-1) total acetic acid compared with values of near zero reported for S. cerevisiae. The inhibitory effects of acetic acid on Z. bailii were not significantly potentiated by ethanol. PMID- 9351275 TI - Production of a cell wall-specific monoclonal antibody to Fibrobacter succinogenes. AB - A monoclonal antibody (mAb) was raised against Fibrobacter succinogenes and produced after fusion as ascites in BALB/c mice. An ELISA was used to test for specificity and sensitivity of the mAb to detect F. succinogenes. The mAb BD1 was tested for sensitivity and cross-reactivity in detecting F. succinogenes with ELISA. The lower limits for F. succinogenes detection in pure and mixed culture using mAb BD1 with ELISA was 10(5) cells ml-1. Twenty-six other species of bacteria, including 12 cellulolytic species, were tested for cross-reactivity with the ELISA but none was detected. Electron micrographs of F. succinogenes cells with immunogold labelling showed that the mAb BD1 reacted exclusively with cell wall epitopes but not intracellular material, as confirmed by ELISA. PMID- 9351276 TI - Heat tolerance of Salmonella typhimurium DT104 isolates attached to muscle tissue. AB - Eight separate experiments were performed with three isolates of Salmonella typhimurium DT104 to examine the impact that attachment to pork muscle tissue has on heat tolerance. In five experiments, attachment to muscle increased heat tolerance. For example, in one experiment the D (58 degrees C) value increased from approximately 2 min for free cells, to > 10 min for attached cells. In three other experiments, differences between free and attached cells were not so pronounced, although attached cells were still more tolerant. This suggests that muscle attachment, which may occur naturally during the preparation of comminuted meat products, could permit greater survival during subsequent cooking and thus may be a possible explanation for the involvement of cooked foods in outbreaks/cases of infection with Salm. typhimurium DT104. PMID- 9351277 TI - Toxicity of crude extracellular products of Aeromonas hydrophila in tilapia, Tilapia nilotica. AB - Extracellular products (ECP) secreted from Aeromonas hydrophila with haemolytic and proteolytic activity were studied with respect to temperature and time of incubation as well as the lethal toxicity on tilapia, Tilapia nilotica. The highest production of the haemolysin product was achieved when Aer. hydrophila was grown at 35 degrees C for 30 h. Tilapia erythrocyte was found to be more susceptible than sheep erythrocyte for determining the haemolytic activity. The haemolytic activity against tilapia erythrocyte was completely inactivated after heating the ECP at 60 degrees C for 10 min or 55 degrees C for 15 min. The proteolytic activity was maximized when the bacterium was grown at 30 degrees C for 36 h. Complete inactivation of the protease enzyme was performed after heating the ECP at 80 degrees C for 10 min or 70 degrees C for 15 min. Aeromonas hydrophila was found to produce haemolytic and proteolytic exotoxin lethal to tilapia (LD50 2.1 x 10(4) cell/fish), as well as heat stable unknown virulent factors that were responsible for 20% mortality. The lethality of ECP was decreased by heating and completely inactivated by boiling at 100 degrees C for 10 min. PMID- 9351278 TI - Interaction of silver nitrate with readily identifiable groups: relationship to the antibacterial action of silver ions. AB - Microbiologically it was demonstrated that amino acids, e.g. cysteine (CySH), and other compounds, e.g. sodium thioglycollate, containing thiol groups neutralized the activity of silver nitrate against Pseudomonas aeruginosa PAO1. Amino acids with disulphide bonds were inactive, with the exception of L-cystine dimethyl ester, as were all amino acids with no sulphur groups. Iodoacetamide reacted with CySH to produce a CyS-acetamide complex that was unable to quench the activity of Ag+. Chemical analyses using cyclic voltammetry demonstrated that high coordination numbers (3.1) were obtained with thiol-containing amino acids and low numbers (0.28-0.4) with other amino acids. Both microbiologically and chemically, the results imply that interaction of Ag+ with thiol groups plays an essential role in bacterial inactivation. PMID- 9351279 TI - Chitinolytic activity of an endophytic strain of Bacillus cereus. AB - Bacillus cereus strain 65, previously isolated as an endophyte of Sinapis, was shown to produce and excrete a chitinase with an apparent molecular mass of 36 kDa. The enzyme was classified as a chitobiosidase because it was able to cleave diacetylchitobiose (GlcNAc)2 from the non-reducing end of trimeric chitin derivatives. The chitinase exhibited activity over the pH range 4.5-7.5 and was stable between pH 4.0 and 8.5. The enzyme had an isoelectric point of 6.4. Application of B. cereus 65 directly to soil significantly protected cotton seedlings from root rot disease caused by Rhizoctonia solani. PMID- 9351280 TI - Flow cytometric analysis of Lactobacillus plantarum to monitor lag times, cell division and injury. AB - Flow cytometry in combination with fluorescent molecular markers 5- (and 6-) carboxyfluorescein succinimidylester (CFSE) and propidium iodide (PI) have been applied to determine lag times, numbers of cell divisions and injury after mild heat (50 degrees C, 5 min) and nisin treatments (0.1 and 1.0 microgram ml-1) of Lactobacillus plantarum. Initial labelling with covalently bound dye CFSE (20 and 100 micrograms ml-1) allowed determination of lag times and cell proliferation for up to eight generations. Double-labelling with CFSE and PI (5 micrograms ml 1) provided additional information about damage levels and distributions within populations. Subpopulations surviving treatment could be identified easily and selectively sorted. PMID- 9351281 TI - An immuno-dot blot assay for detection of thermostable protease from Pseudomonas sp. AFT-36 of dairy origin. AB - A dot-ELISA technique for the detection of Pseudomonas protease was developed using IgG of anti-Pseudomonas AFT-36 protease as capture antibody. The detection limit of protease in buffer or milk was 1.01 ng ml-1. The procedure was performed at room temperature, took about 2.5 h and was economical. Protease AFT-36 is immunologically related to five out of seven Pseudomonas spp. The results suggest that the assay could be used to detect proteases in dairy products. PMID- 9351282 TI - PCR amplification of crude microbial DNA extracted from soil. AB - A rapid, inexpensive, large-scale DNA extraction method involving minimal purification has been developed that is applicable to various soil types. DNA was extracted from 100 g of soil using direct lysis with glass beads and sodium dodecyl sulphate (SDS) followed by polyethylene glycol precipitation, potassium acetate precipitation, phenol extraction and isopropanol precipitation. The crude extract could be used in PCR directed at high-copy number (bacterial small subunit rRNA) and single-copy (fungal beta-tubulin) genes. PMID- 9351283 TI - Biopolymers from marine prokaryotes. AB - Biopolymers from marine prokaryotes, both Bacteria and Archaea, offer a number of novel material properties and commercial opportunities. The characteristics of marine exopolysaccharides and melanins that enhance the survival ability of the organisms producing them can be exploited for a number of products ranging from emulsifiers to adhesives. In the prokaryotes, the polyhydroxyalkanoates form carbon-storage molecules, but their technological application is entirely different, serving as a potential base material for biodegradable plastics. Marine biopolymers are a significant and undeveloped biological resource. PMID- 9351284 TI - Polyunsaturated fatty acids, Part 1: Occurrence, biological activities and applications. AB - Polyunsaturated fatty acids form a unique class of food constituents that show a wide range of functions in biological systems. Investigations over the past two decades have uncovered their roles and those of their eicosanoid metabolites, and have highlighted their homeostatic functions in mammals. A growing number of common human medical conditions are thought to be traceable to dysfunctions in the eicosanoid system, which could in turn be due to imbalances in the intake and/or metabolism of polyunsaturated fatty acids. This, together with medical advances, has spurred the introduction of biomedical products, nutritionals, fortified foods and health supplements. PMID- 9351285 TI - Drug delivery to the nervous system. AB - Delivery of drugs to the nervous system remains a challenge despite advances in our understanding of the mechanisms involved in the development of neurodegenerative disorders and the actions of neuroactive agents. Drug accessibility to the central nervous system is limited by the blood-brain barrier; although the peripheral nervous system is more accessible than the central nervous system, problems are still encountered, mainly owing to the poor stability and considerable side effects of many neuroactive compounds when administered systemically. Microencapsulation of neuroactive compounds and living cells producing such substances can overcome some of these shortcomings for delivery to the nervous system. PMID- 9351286 TI - Emerging tandem-mass-spectrometry techniques for the rapid identification of proteins. AB - State-of-the-art techniques such as liquid-chromatography-electrospray-ionisation tandem mass spectrometry have, in conjunction with database-searching computer algorithms, revolutionised the analysis of biochemical species from complex biological mixtures. With these techniques, it is now possible to perform high throughput protein identification at picomolar to subpicomolar levels from protein mixtures. This article provides an overview of the techniques and methodologies available for the structural elucidation and identification of proteins and peptides from complex biological samples. PMID- 9351287 TI - Applications of dielectrophoresis in biotechnology. AB - Recent progress in the development of microelectrode structures has led to new techniques for the dielectrophoretic characterization and sorting of cells, microorganisms and other bioparticles using nonuniform AC electric fields. These methods utilize differences in the dielectric polarizabilities of cells for their effectiveness, and factors controlling such properties include the conductivity and permittivity of membranes and any cell walls, electrical double layers associated with surface charges, cell morphologies, and internal structures. Applications of dielectrophoresis have included the selective spatial manipulation and separation of mixtures of bacteria, viable and unviable cells, cancerous and normal cells, and red and white blood cells. PMID- 9351288 TI - Membrane-protein engineering. AB - Membrane proteins perform many of the essential functions required for life. They are often the targets of medicinal drugs and have many potential uses in biotechnological processes. Therefore our ability to understand them and manipulate their functions is both important and necessary to enable protein engineers to create 'designer' membrane proteins (that is, proteins designed to have desired properties). PMID- 9351289 TI - Advances in immunotherapy. Cellular and molecular bases of modern immunotherapy. PMID- 9351290 TI - New immunosuppressive agents for organ transplantation. PMID- 9351291 TI - Neonatal sepsis due to nonencapsulated Haemophilus influenzae biotype IV. AB - A case report of a newborn with sepsis due to nontypable H.Influenzae biotype IV is presented. There were no prematurity nor maternal obstetrical complications involved. The child however suffered from severe respiratory distress. With the aspiration of secretions, the resuscitation with mask oxygen and the empirically started combination of ampicillin and cefotaxime, his condition rapidly improved. PMID- 9351292 TI - Palliative care: knowledge and attitudes of general practitioners: the results of a questionnaire after training. AB - A questionnaire about attitudes and knowledge in palliative care treatment was sent to 185 general practitioners who participated such a seminar some months before. The response rate was low (69/185). Pain is the most frequent symptom. All the responding doctors assume they can treat pain adequately. However, none of them can answer the knowledge questions correctly. They appreciate the palliative care organizations for the help in symptom control or for difficulties in psychological symptoms. A multifactorial etiology might explain the difficult control of asthenia. This survey agrees with other studies that knowledge in symptom control and palliative care should be improved. PMID- 9351293 TI - Pressure recovery across the aortic valve. AB - The Bernoulli equation relates the pressure exerted on a fluid to its flow velocity and its density, in addition to its flow acceleration and its viscous friction loss. When flow velocity increases at a narrowing, the local pressure decreases proportionally. It has been wrongfully assumed that pressure lost distal to a stenosis can never recover. It is, however, the energy content of the fluid, equal to the kinetic plus the potential energy, which can not increase. When flow slows distal to a narrowing and little energy is lost to friction, pressure does actually increase. Pressure recovery has been well demonstrated to exist in a variety of pathophysiological states. Bicuspid aortic valve prostheses such as the St. Jude valves can produce quite remarkable pressure recovery. This causes a great discrepancy between pressure drop calculations based on continuous wave doppler on the one hand and true pressure drop across the prosthesis on the other. Reliance on doppler measurements only might wrongfully lead one to conclude that the prosthesis was malfunctioning. Less extreme pressure recovery is possible across a stenotic native aortic valve, but interpretation of the flow velocity across the valve might make the difference between recommendations to replace or to retain the valve. When interpreting doppler signals across narrowings the phenomenon of pressure recovery should be kept in mind. PMID- 9351294 TI - Total quality management for clinical laboratories: a need or a new fashion? AB - In most European countries, concepts of quality management in medical laboratories have been proposed. These concepts are based on general standards for test laboratories (EN 45001, ISO 25) or specific adapted standards. Improvement of quality lays on the foundation of the implementation of quality systems in medical laboratories. This new approach will have consequences on management style and on working conditions. Efficacy on the implementation can only be tested by external audits. During this audit, not only the quality system and analytical competence must be examined, but also if there is a real contact between pathologists and clinicians and if laboratory results are clinically validated (clinical audit). This new vision on quality in medical laboratories will ask a reconsideration of the tasks, duties and knowledge of clinical pathologists. PMID- 9351295 TI - Recent advances in Alport syndrome: the cross-fertilisation between genetics and clinical medicine. PMID- 9351296 TI - Hypoglycemia or non-hypoglycemia. PMID- 9351297 TI - "How do you do it?". PMID- 9351299 TI - Validity of the TONI-2 with deaf and hard of hearing children. AB - The test of Nonverbal Intelligence, 2nd edition (TONI-2) is a quick, practical measure of cognitive functioning. Its concurrent, construct, and predictive validity as an instrument and its utility with deaf and hard of hearing children were explored in this study. Results obtained from research with 27 deaf and hard of hearing children of elementary school age indicated a positive, moderate correlation between the TONI-2 IQ and the Wechsler Intelligence Scale for Children, 3rd edition (WISC-III), Performance IQ, supporting concurrent validity. Construct validity was established by significant correlations between the TONI-2 and five of six WISC-III subtests. The TONI-2 had predictive value for Stanford Achievement Test (SAT) spelling and mathematical problem-solving and reasoning tasks. PMID- 9351298 TI - Support services for parents and their children who are deaf or hard of hearing. A national survey. AB - In the spring and summer of 1996, 404 parents of children who are deaf or hard of hearing and were born in 1989 or 1990 completed questionnaires about early services received and the current status of their children's development. The children were enrolled in 137 different programs in 39 states; about one quarter of the programs participating in Gallaudet's Annual Survey of Deaf and Hard-of Hearing Children and Youth. Children who are deaf comprised 46% of the group for whom responses were provided; those who are hard of hearing comprised 54%. Hearing loss was confirmed at the mean age of 14.5 months for the deaf group, and at the mean age of 28.6 months for the hard of hearing group. Additional conditions place 32% at risk for educational or developmental difficulties. One or both parents are deaf in 13% of responding families. Almost 40% of mothers have some training beyond high school; one third of the children came from non White or mixed-race backgrounds. (These characteristics of children and families are used in analyzing other data collected from responding parents). Communication approaches used in children's initial programs included: speech alone (24%), sign + speech (66%), sign alone (5%), cues (3%), and sign + cues (3%). Parents gave highly favorable evaluations to intervention programs, and placed teachers at the top of a "sources of help" list. Parents from minority groups and those with no college training reported that their children showed more behavior problems and less language progress, and gave more negative responses to questions regarding the impact of deafness on their families. This suggests that program personnel may need to increase their intervention efforts for these subgroups of special education consumers. PMID- 9351300 TI - Pedophilia and deafness. AB - Data from 22 cases of deaf individuals suffering from pedophilia are presented along with a tabular summary of recent articles from the deaf press, about deaf victims of pedophilia and deaf pedophiles. Results indicate a number of factors that distinguish deaf pedophiles from hearing pedophiles. First is the prevalence of Primitive Personality Disorder in the deaf group. Corollary to this, with a significant number of pedophiles, competence to stand trial is a major issue. Other significant differences include a high rate of brain damage, illiteracy, poor communication skills, and other psychiatric illnesses. Two of the 22 cases were deaf females with pedophilia. The mean performance IQ of the sample was 102.8 and the distribution of scores was bimodal. Case histories are presented and discussed, and legal issues, prevention, and punishment are addressed. PMID- 9351301 TI - Using a TTY to contact clinical graduate programs. AB - This study investigated accessibility of 177 APA-accredited clinical and counseling programs to deaf applicants via TTY phone lines, what happened when we called these numbers, and where these phone lines connected. We were able to obtain TTY phone numbers for 135 schools, of which we could successfully reach 86 schools using a TTY alone. Most of these lines (60%) were connected to campus disabled student services, and none connected directly to the APA-accredited programs or the departments in which they were housed. Comments from university personnel underscore the difficulties facing deaf applicants. We argue that the difficulties deaf applicants encounter when trying to contact programs constitute significant barriers to the application process. We give six specific recommendations for expanding access to programs. PMID- 9351302 TI - A group storybook-reading intervention with children at a residential school for the deaf. AB - Deaf children's patterns of emergent literacy can parallel those of hearing children, but their acquisition of conventional reading skills in elementary school is often delayed. Group storybook reading in the residences of a state sponsored school for the Deaf was investigated as a means of fostering literacy development outside the classroom. Eighteen children, ages 4-11 years, participated. The nine children in the experimental group cottages participated in group storybook reading twice each week for 5 months. Both the experimental and the control-group cottages were provided with a variety of books that were rotated biweekly. Children were highly engaged during the storybook-reading sessions, particularly when the readers used an interactive/expressive reading style. Children in the experimental group performed more independently on an emergent reading task, and their counselors judged them to be more interested in books than children in the control group. PMID- 9351303 TI - The use of signed English pictures to facilitate reading comprehension by deaf students. AB - Many deaf students have severe difficulty acquiring literacy and developing reading comprehension beyond an elementary school level. This difficulty apparently results from a combination of perceptual, communication, instructional, linguistic, and experiential deficits. Although some deaf students develop a degree of signed English proficiency, this does not necessarily translate into reading proficiency. Recent studies examining the possible association between signed English pictures and comprehension of printed text present some support for facilitation of students' word recognition in a format combining those two elements. Whether this format enhances comprehension remains unclear from previous studies. The present study, involving 16 severely or profoundly deaf students across two reading-proficiency groups, examined whether the use of signed English pictures in association with printed text enhances students' reading comprehension. The study found that comprehension was significantly enhanced by the use of signed English reading books, with poorer readers deriving greater benefit than better readers. Practical implications of the findings are discussed. PMID- 9351304 TI - Schwannoma of the trachea. AB - In this study we present a case of schwannoma of the trachea investigated with computer tomography (axial, precontrast) and magnetic resonance (1 Tesla, sagittal T1w before and after Gadolinium, sagittal T2w, coronal and axial after Gadolinium). The topographic abilities of MR allowed us to determine the exact location and extension of the tumor in the trachea. As far as we know, this study, based on magnetic resonance, is the first of the kind in the radiologic literature. PMID- 9351305 TI - Ischemic jejunitis caused by primary small bowel volvulus. AB - A case of ischemic jejunitis caused by primary small bowel volvulus is presented. The radiological signs of ischemia persisted after detorsion. Contrast examinations of the small intestine demonstrated severe jejunitis with ulcerations, segmental narrowing and fistulas. This last sign is a rather uncommon event in ischemic disorders. The radiological signs of acute ischemia of the small intestine are discussed. PMID- 9351306 TI - Is intravenous urography still used in patients with prostatism? AB - The value of intravenous urography in patients with prostatism was retrospectively evaluated. One thousand four hundred ninety five intravenous urograms of male patients referred by the department of urology were reviewed. Based on the clinical information, only patients with complaints of prostatism as a single symptom were selected. Patients with associated symptoms (i.e. hematuria, urinary infection) were excluded. Forty seven patients could be included based on these criteria. In 29 of 47 cases (61.7%) no abnormalities were found. Abnormalities found in 18 cases included dilatation of the excretory system, urinary calculi, congenital anomaly, acquired small kidney, renal cysts and retroperitoneal fibrosis. In 5 cases (10.1%) the intravenous urography necessitated further treatment and/or follow up. In 3.1% prostatism was the indication for the examination. The number of relevant abnormalities at intravenous urography performed for prostatism is low and this is in accordance with results reported in literature. These results provide further evidence for the continuously changing indications for intravenous urography. PMID- 9351307 TI - Dural sinus thrombosis: CT and MR imaging of different stages. AB - Cerebral dural sinus thrombosis remains an uneasy clinical diagnosis because it may present with a spectrum of nonspecific manifestations. CT and MR findings have been described to help recognize this entity. We report here a case with different stages of thrombosed superior sagittal and right transverse dural sinuses demonstrated by CT and MR imaging. PMID- 9351308 TI - Mycotic pseudo-aneurysm of the extracranial carotid artery. AB - A rare case of mycotic pseudo-aneurysm of the common carotid artery as a complication in an immunosuppressed paediatric patient is presented. Treatment of pseudo-aneurysms of the common carotid artery is generally considered to be an emergency, necessitating quick and accurate diagnosis. In patients with septicemia, angiography has to be avoided. We were able to provide the surgeon with the exact diagnosis and accurate topographical information with helical CT with 3D reformation. PMID- 9351309 TI - A patient with Peyronie's disease: ultrasonographic features. AB - Clinical, radiographic, and ultrasonographic findings in a 45-year-old patient with Peyronie's disease are reported. Ultrasonography precisely identified calcified as well as fibrous plaques characteristic of this disorder. PMID- 9351310 TI - Ultrasonography of the knee. AB - A comprehensive review of ultrasonography of the knee is presented and includes the choice of equipment and best patient positioning as well as a review of the normal anatomy and major disorders involving the knee, menisci and ligaments. PMID- 9351311 TI - Low cost MR imaging: medical and economic perspectives. AB - A critical review of the potential applications, clinical usefulness and shortcomings of low field MR imaging with a view to cost control is provided. PMID- 9351312 TI - Facet joints diseases. AB - A review of the normal anatomy, developmental abnormalities, traumas and disorders affecting the facet joints is provided, with a recall of the major imaging modalities. PMID- 9351313 TI - Digital imaging: the Agfa experience. PMID- 9351314 TI - 1997 William J. Stickel Gold Award. Morphological and biochemical properties of metatarsophalangeal joint cartilage. AB - Although there is sparse information concerning the properties of foot-joint cartilages, knowledge of the morphology and biochemistry of these cartilages is important in the study of changes that occur in the development of osteoarthritis. Normal first and fifth metatarsophalangeal joints were chosen for comparison because of the difference between these two joints in the prevalence of osteoarthritis, particularly with advancing age. The authors' study shows that there is no age-related decrease in articular-cartilage thickness; however, there is an age-related decrease in the chondrocyte density in the superficial zone in both joints. There is, however, a difference between the two joints in the level of expression of matrix-degrading enzymes. This difference may indicate differences in specific chondrocyte activity that precedes or accompanies the development of osteoarthritis or other degenerative morphological changes. PMID- 9351315 TI - 1997 William J. Stickel Silver Award. The anti-inflammatory action of locally injected ketorolac. AB - Locally injected steroids are used to treat inflammatory conditions, in spite of the complications associated with their use. Ketorolac tromethamine, an injectable nonsteroidal anti-inflammatory drug, has not previously been evaluated for treatment of musculoskeletal inflammatory conditions via local administration. Eighty Achilles tendons of rabbits were traumatized in a controlled fashion. At the time of trauma, a single dose of ketorolac (1, 3, or 5 mg/kg) or normal saline was administered peritendinously. Three days later, the tendons were harvested and examined histologically to evaluate the degree of inflammation present in the tissue. No statistically significant difference was found between the experimental and control groups. The authors conclude that locally injected ketorolac does not prevent the onset of an inflammatory process. PMID- 9351316 TI - 1997 William J. Stickel Bronze Award. Comparison of strategies for reducing pressure at the site of neuropathic ulcers. AB - Few scientific data are available on the effectiveness of commonly used modalities for reducing pressure at the site of neuropathic ulcers in persons with diabetes mellitus. The authors' aim was to compare the effectiveness of total contact casts, half-shoes, rigid-soled postoperative shoes, accommodative dressings made of felt and polyethylene foam, and removable walking casts in reducing peak plantar foot pressures at the site of neuropathic ulcerations in diabetics. Using an in-shoe pressure-measurement system, data from 32 midgait steps were collected for each treatment. There was a consistent pattern in the devices' effectiveness in reducing foot pressures at ulcer sites under the great toe and ball of the foot. Removable walking casts were as effective as or more effective than total contact casts. Half-shoes were consistently the third most effective modality, followed by accommodative dressings and rigid-soled postoperative shoes. PMID- 9351317 TI - Dilute lidocaine ankle blocks in the diagnosis of sympathetically maintained pain. AB - The author has developed a technique of dilute anesthetic ankle block that appears, on the basis of these preliminary observations, to relieve pathologic pain that may be maintained by the sympathetic nervous system. Symptomatic relief following the use of this injection confirms that the patient's problem is not somatic and that further evaluation and treatment of sympathetically maintained pain syndrome may be indicated. PMID- 9351319 TI - Ankle arthrodesis following avascular necrosis of the talus in a patient with lupus. AB - Avascular necrosis of bone is a common manifestation of systemic lupus erythematosus, particularly in those patients receiving corticosteroids. The authors review the pathogenesis and diagnosis of avascular necrosis and describe an ankle arthrodesis in a patient with systemic lupus erythematosus who developed avascular necrosis of the talus. PMID- 9351318 TI - Mixed cavernous and capillary intraosseous hemangioma of the foot. AB - Hemangiomas of bone are rare lesions accounting for approximately 1% of all primary bone tumors. Intraosseous hemangiomas of the foot are especially rare, with only sparse reports in the literature. Presented here is a case study of an erosive bony lesion of the midfoot that was microscopically and histopathologically proven to be a mixed cavernous and capillary hemangioma. Eradication of the lesion during diagnostic biopsy obviated further treatment. PMID- 9351320 TI - Torsion of the tendon of peroneus brevis. AB - The authors present a previously undescribed torsion located within the tendon of peroneus brevis. The musculotendinous unit of peroneus brevis was isolated from 46 lower extremities of cadavers. A goniometer was constructed and utilized to quantify the degree of torsion located within each peroneus brevis tendon. Torsion was present in all 46 cadaver specimens, with a mean of 38.5 degrees and a range of 26 degrees to 56 degrees. The regional anatomy and biomechanical functions of peroneus brevis are discussed, and proposed bases for the embryologic origins and functional significance of the torsion are presented. PMID- 9351321 TI - Using humor in medical practice. PMID- 9351322 TI - Screening for hemochromatosis encouraged. PMID- 9351323 TI - Radiology Quiz #14. Spleen laceration and bowl rupture. PMID- 9351324 TI - Independent physician associations (IPAs): regaining control of managed care. PMID- 9351325 TI - The use of standard management reports in medical practice. Part II. PMID- 9351326 TI - Clinical predictors of domestic violence. PMID- 9351327 TI - The causes of hemoptysis revisited. A review of the etiologies of hemoptysis between 1986 and 1995. PMID- 9351328 TI - Pathophysiology of liver circulation with an overview of medical and invasive treatments. AB - Liver hemodynamics is characterized by a dual venous and portal blood supply whose physiologic variations are particularly evident during digestion. In the normal subject portal blood flow is laminar with the left liver receiving the blood from the small intestine while the left liver is supplied by the blood from the spleen and colon. In pathologic conditions increased arterial blood flow accompanies the decreased portal flow. Portal hypertension in its various forms is the most frequent and important circulatory alteration in chronic liver disease; besides the "organic" obstacles to the hepatic blood flow there are the dynamic mechanisms which regulate the vascular resistance in the microcirculation Therapies which impact on liver circulation are surgical, of interventional radiology and medical, used in the prevention and cure of complications of portal hypertension. PMID- 9351329 TI - Role of angiography in the evaluation of hepatic perfusion. AB - Angiography was the first method to be used for a morphofunctional study of hepatic perfusion. It can be performed with direct puncture of portal system or indirect opacification after contrast injection into the splenic artery or superior mesenteric artery. At present, direct angiographic procedures have only a historical value in the diagnostic approach while they have a preliminary role in interventional maneuvers on the portal system (TIPSS, embolization of portal branches or left gastric vein). Indirect angiographic procedures allow the study of arteries, parenchymas and portal system. Much of the information on arterial hepatic and portal circulation is now supplied by noninvasive procedures (US,CT,MRI); however in selected cases, angiography can be performed. Furthermore, the knowledge of angiographic findings of hepatic circulation is basic to the interpretation of "functional" information supplied by color-Doppler US, spiral CT and MR angiography. PMID- 9351330 TI - Is there still a role for functional radionuclide study of the liver? AB - Clinical applications of radionuclide methods for the study of liver hemodynamics and hepatocyte function are examined. In particular, as for hemodynamic studies, perfusion assessment with radiocolloids, 99mTc-IDA scintigraphy or 99mTc-labeled red blood cells, is underlined; they allow characterization of the different cellular component of space-occupying liver processes. The use of hepatic perfusion index (HPI) is reconsidered both as prognostic parameter in cirrhotic patients and as predictor of liver metastasis from colorectal cancer. The diagnostic role of recent procedures, as those based on endorectal radiopharmaceuticals in the evaluation of portosystemic shunts in cirrhosis, is analyzed. Studies of hepatocyte function of practical concern are essentially devoted to the "excretory function" and "asialoglycoprotein metabolism". In the first case, a major role is played by IDA halogenated derivatives and functional parameters drawn from them by mathematico-statistical evaluations of radiohepatogram (simple or applied to compartmental models). For metabolic studies, at present an artificial glycoprotein, 99mTc-galactosylneoglycoalbumin (99mTc-NGA) that binds with hepatocellular receptors is used. Information on the rate of blood plasma clearance and liver uptake, receptor density (altered in some pathologic conditions) and plasmic hepatic flow, is supplied. PMID- 9351331 TI - Color Doppler US of intrahepatic vascular system. AB - Aim of this article is an up-dating of the state of the art of color Doppler US in the assessment of intrahepatic vascularization. Recent reports are reviewed, based on already acquired certainties to better the knowledge of the physiology and pathophysiology of hepatic circulation to investigate new clinical applications of color Doppler US. PMID- 9351332 TI - Hepatic perfusion: new perspectives at computed tomography. AB - The dual blood supply to the liver, that of hepatic arterial supply and portal venous blood supply represents both opportunity and trouble for radiologists in using contrast material to detect liver neoplasms. It is this unique feature that allows one to optimize tumor detection by optimizing contrast flow to either the liver parenchyma or to tumors. Conversely, however, if one does not optimize contrast administration to take advantage of differences in blood flow between liver parenchyma and liver neoplasms, potentially tumors can be obscured at imaging. The focus of this article will be to review the physiology of liver blood flow and how various methods of contrast material administration can be used to optimize CT liver tumor detection and characterization. PMID- 9351334 TI - Diffuse liver disease: anatomopathologic indications for functional radiology of the liver. AB - Main histopathologic features of diffuse liver disease with particular reference to chronic viral hepatitis in view of functional radiology of the liver are presented and discussed. Attention is focused on histological grading and staging of fibrosis in chronic hepatitis, considered as most significant histopathologic parameters for comparison with the findings of functional radiology. PMID- 9351333 TI - Functional radiology of the liver: magnetic resonance imaging. AB - Magnetic Resonance (MR) images sensitive to the flowing blood are defined as images of MR angiography. Proton movement within a magnetic field modifies both the intensity and the phase of Nuclear Magnetic Resonance (NMR) signal; two techniques of MR angiography are thus distinguished: (TOF) the "time of flight" (intensity) and the "phase-contrast" (phase) technique. In the time of flight MR angiography the blood may appear as hypointense or hyperintense compared to stationary tissues. Blood hypointensity in vessels is due to the flow void phenomenon while hyperintensity is due to the phenomenon of flow-related enhancement. In phase contrast MR angiography, protons moving within a magnetic field modify their phase directly proportional to the displacement velocity and gradient intensity. Moreover, MRI allows noninvasive measurement of blood flow. Flow velocity is measured with TOF sequences or phase-contrast sequences. In TOF sequences quantitative measurement is performed with the bolus tracking procedure. In contrast-phase sequences the velocity is measured based on the extent of signal phase modification induced by the proton displacement velocity. The recent use of liver-specific contrast media supplies information on parenchymal liver function. PMID- 9351336 TI - Hares and tortoises in the race to sequence the human genome: expectations and realities. PMID- 9351335 TI - New perspectives on the clinical applications of functional radiology of the liver. AB - Preliminary results of "new clinical applications" of functional imaging of the liver are reported. In 20 healthy volunteers portal flow measurement with Doppler US at the level of right, left portal branch and main portal trunk, showed the preferential distribution in baseline conditions of portal flow to the right liver (about 68%) as compared to the left portal branch. This influenced MRI volumetry of right liver as compared to left liver. After meal intake, flow increase was significantly higher at the level of left portal branch suggestive for a "functional reserve" in left liver. Portal flow physiology was examined in preparation of portal imaging before and after portal vein embolization, a procedure performed preoperatively before enlarged hepatectomies. PMID- 9351337 TI - A TWIST in development. PMID- 9351338 TI - Executive decision: chromatin structure and gene regulation. PMID- 9351339 TI - Differential genome display. PMID- 9351340 TI - Comparative genomics: lessons from cats. AB - The genomics era, spear headed by dazzling technological developments in human and mouse gene mapping, has additionally provoked extensive comparative gene mapping projects for domestic species of several vertebrate orders. As the human genome project promises a one dimensional string of 100,000 genes and sequences, comparative mapping will extend that inference to a second dimension representing index species of the 20 living mammalian orders and to a third dimension by phylogenetic description of the genomes of mammal ancestors. We review here the remarkable extent of genome homology conservation among mammals illustrated by technology applications in the feline genome project. PMID- 9351341 TI - Haemophilus influence: the impact of whole genome sequencing on microbiology. AB - The publication of the Haemophilus influenzae genome sequence in 1995 was a landmark in microbiological research. It has changed our understanding of the prokaryotic world, and will influence the approach and focus of research on microorganisms over the next few years. In this article we outline what has been learned from this and other genome sequencing projects, and discuss some of the potential avenues of investigation that will follow in the 'post-genome era'. PMID- 9351342 TI - Genome screening by comparative genomic hybridization. AB - Comparative genomic hybridization (CGH) provides a molecular cytogenetic approach for genome-wide scanning of differences in DNA sequence copy number. The technique is now attracting wide-spread interest, especially among cancer researchers. The rapidly expanding database of CGH publications already covers about 1500 tumors and is beginning to reveal genetic abnormalities that are characteristic of certain tumor types or stages of tumor progression. Six novel gene amplifications, as well as a locus for a cancer-predisposition syndrome, have been discovered based on CGH data. CGH has now been established as a first line screening technique for cancer researchers and will serve as a basis for ongoing efforts to develop high-resolution next-generation genome scanning, such as the microarray technology. PMID- 9351343 TI - Sunlight and the onset of skin cancer. AB - The photons of sunlight precipitate a series of genetic events in skin leading to cancer. These events involve somatic mutations as well as inherited alleles. Competition between cell populations ensues, as a single mutated cell expands into a clone. Thus cancer involves both a single-cell problem and a many-cell problem; in skin cancer, sunlight appears to drive both. PMID- 9351344 TI - SWISS-PROT + TREMBL. PMID- 9351345 TI - Nitric oxide synthases and cardiac muscle. Autocrine and paracrine influences. AB - The different cell types comprising cardiac muscle express one or more of the three isoforms (neuronal NOS, or nNOS; inducible NOS, or iNOS; and endothelial NOS, or eNOS) of nitric oxide synthase (NOS). nNOS is expressed in orthosympathetic nerve terminals and regulates the release of catecholamines in the heart. eNOS constitutively expressed in endothelial cells inhibits contractile tone and the proliferation of underlying vascular smooth muscle cells, inhibits platelet aggregation and monocyte adhesion, promotes diastolic relaxation, and decreases O2 consumption in cardiac muscle through paracrinally produced NO. eNOS is also constitutively expressed in cardiac myocytes from rodent and human species, where it autocrinally opposes the inotropic action of catecholamines after muscarinic cholinergic and beta-adrenergic receptor stimulation. iNOS gene transcription and protein expression are induced in all cell types after exposure to a variety of inflammatory cytokines. Aside from participating in the immune defense against intracellular microorganisms and viruses, the large amounts of NO produced autocrinally or paracrinally mediate the vasoplegia and myocardial depression characteristic of systemic immune stimulation and promote cell death through apoptosis. In cardiac myocytes, NO may regulate L-type calcium current and contraction through activation of cGMP dependent protein kinase and cGMP-modulated phosphodiesterases. Other mechanisms independent of cGMP elevations may operate through interaction of NO with heme proteins, non-heme iron, or free thiol residues on target signaling proteins, enzymes, or ion channels. Given the multiplicity of NOS isoforms expressed in cardiac muscle and of the potential molecular targets for the NO produced, tight molecular regulation of NOS expression and activity at the transcriptional and posttranscriptional level appear to be needed to coordinate the many roles of NO in heart function in health and disease. PMID- 9351346 TI - The unstable atheroma. PMID- 9351347 TI - Cellular heterogeneity of the vascular tunica media. Implications for vessel wall repair. PMID- 9351348 TI - Redistribution of von Willebrand factor in porcine carotid arteries after balloon angioplasty. AB - von Willebrand factor (VWF) is a well-characterized multimeric glycoprotein present in platelets and plasma and synthesized by vascular endothelial cells and megakaryocytes. Its role in platelet-vessel wall interactions has been studied extensively, but its involvement in intravascular events after balloon angioplasty has not been clarified. VWF antigen is not present in porcine arterial endothelium (except for the pulmonary artery) but is readily detected in porcine venous endothelial cells. We have examined the localization of VWF in porcine vessel walls during neointima formation after bilateral carotid balloon angioplasty. Endothelium was denuded by balloon injury but regenerated by 7 days and was fully confluent by 42 days. VWF was detected at the site of injury in localized, adherent platelet aggregates at 10 minutes after angioplasty that were not present at later time points. A well-demarcated homogeneous layer of VWF was observed on the luminal surface from 30 minutes to day 7, but there was a progressive shift of positive staining from the lumen to the outer media from days 1 to 7. VWF was also strongly detected at sites proximal and distal to the balloon injury from 30 minutes to day 7, although endothelial disruption was minimal and the monolayer remained substantially intact at these sites. Regrowing endothelial cells appeared to contain granular VWF from days 12 to 21, but this was not readily evident at later time points. The results suggest that balloon injury is associated with deposition and medial absorption of plasma or platelet VWF in this porcine model over a time period that precedes and overlaps vascular smooth muscle proliferation and endothelial recoverage. The findings provide evidence to support the concept of a wider role for VWF in tissue injury responses. PMID- 9351349 TI - Hepatic lipase gene polymorphisms influence plasma HDL levels. Results from Finnish EARS participants. European Atherosclerosis Research Study. AB - Hepatic lipase (HL), a triglyceride lipase found in liver, adrenals, testes, and ovaries, takes part in the uptake, remodeling, and function of lipoproteins including HDL, as well as VLDL and chylomicrons. In the present study, the genotype distribution of five HL polymorphisms (-C480T, V133V, T202T, L334F, T457T) and their association to plasma lipid values were investigated. The study participants included 92 students with paternal history of myocardial infarction before the age of 55 and 194 matched control subjects, ie, the Finnish participants of the European Atherosclerosis Research Study (EARS). The allele T of the HL polymorphism -C480T showed an association with elevated HDL, apoA-I, and LpA-I values (ANOVA P < .01). No difference in genotype distribution was observed in the offspring with and without paternal history of myocardial infarction. PMID- 9351350 TI - Normolipidemic subjects with low HDL cholesterol levels have altered HDL subpopulations. AB - Epidemiological studies have established that plasma concentration of HDL is inversely correlated with the risk of coronary heart disease, even in the absence of increased LDL cholesterol levels. We postulate that specific HDL subpopulations may be responsible for antiatherogenic properties of HDL. HDL subpopulations were quantitated by two-dimensional gel electrophoresis in 79 normolipidemic healthy male subjects. To eliminate the influence of diet, volunteers consumed an average American diet for 6 weeks. After the diet period, subjects were stratified according to their HDL cholesterol (HDL-C) levels to low HDL-C < 0.91 mmol/L (< 35 mg/dL), medium > 0.91 < 1.30 mmol/L (> 35 < 50 mg/dL), and high > or = 1.30 mmol/L (> or = 50 mg/dL) groups. Plasma triglycerides and insulin levels were in the normal range, but subjects with low HDL-C levels had higher concentrations of plasma triglycerides and insulin than subjects with medium or high HDL-C concentrations. The absolute concentration (mg/dL) of apoA-I in the largest alpha-migrating HDL subpopulation (alpha 1) was (P < .01) lower in the low HDL-C subjects compared with the medium and high HDL-C groups. The relative concentration (percent distribution) of apoA-I was decreased (P < .01) in alpha 1 and increased (P < .01) in alpha 3 subpopulations. A positive correlation between HDL-C and alpha 1 (P < .001) and a negative correlation between HDL-C and alpha 3 were observed. The inverse correlation of apoA-I distribution (relative concentration) between alpha 1 and alpha 3 suggests an interconversion of alpha 1 and alpha 3 subpopulations, possibly by cholesteryl ester transfer protein. Pre-beta subpopulations showed an inverse trend with HDL C, while the pre-alpha subpopulation behaved similarly to the alpha-migrating subpopulation. Colocalization of apoA-I and apoA-II particles in the different HDL subpopulations demonstrated that alpha 1, pre-beta 1, and pre-beta 2 subpopulations are apoA-I-only particles rather than apoA-I:A-II particles. PMID- 9351351 TI - Excess prevalence of fasting and postmethionine-loading hyperhomocysteinemia in stable renal transplant recipients. AB - Hyperhomocysteinemia, either fasting or after methionine loading, may contribute to the increased incidence of cardiovascular disease events experienced by renal transplant recipients. Limited data are available on fasting homocysteine (Hcy) levels, and none on postmethionine-loading Hcy levels, in these patients. We assessed the prevalence and potential determinants of fasting and postmethionine loading hyperhomocysteinemia in 29 stable renal transplant recipients and 58 age- and sex-matched, population-based controls free of renal disease with serum creatinine levels of 1.5 mg/dL or less. Total (t) plasma Hcy was determined fasting and 2 hours after methionine loading, along with fasting determinations of the B-vitamin cofactors/substrates for Hcy metabolism, ie, pyridoxal 5' phosphate, B-12, and folate and serum creatinine. Geometric mean fasting (18.1 versus 9.8 microM, P < .001) and postmethionine-loading increase (22.0 versus 15.2, P = .001) in tHcy levels were significantly greater in the renal transplant recipients, as were the prevalence odds (with 95% confidence intervals) for fasting [14.8 (3.4-64.7)], postmethionine loading [6.9 (1.5-32.8)], combined fasting and postmethionine-loading [18.0 (2.3-142.1)] hyperhomocysteinemia, and inadequate circulating folate [4.2 (1.1-16.5)] or pyridoxal 5'-phosphate [3.2 (0.9-11.0) status. Correlation analyses suggested important potential relationships between creatinine and both fasting (+0.64, P < .001) and postmethionine-load increase (+0.38, P = .045) in tHcy, folate and fasting ( 0.41, P = .025) tHcy, and pyridoxal 5'-phosphate and postmethionine-loading increase (-0.33, P = .091) in tHcy. We conclude that there is an excess prevalence of fasting and postmethionine-loading hyperhomocysteinemia in stable renal transplant recipients. Renal function is related to both fasting and postmethionine loading-hyperhomocysteinemia, inadequate folate status is associated with fasting hyperhomocysteinemia, and inadequate vitamin B-6 status may be related to postmethionine-loading hyperhomocysteinemia in this patient population. PMID- 9351354 TI - Factor VII polymorphisms in populations with different risks of cardiovascular disease. AB - Increased plasma factor VII coagulant activity (FVII:C) has been associated with the risk of ischemic heart disease (IHD). Differences in plasma FVII:C among individuals are associated with three common polymorphisms in the FVII gene. Therefore, we investigated FVII polymorphisms in four populations that differ in their risk of developing cardiovascular disease, namely, Europeans, Greenland Inuit, Gujarati Indians, and Afrocaribbeans. We studied (1) the promoter polymorphism, which is the result of a decanucleotide insertion in the FVII promoter at position -323 from the start of translation; (2) the hypervariable region 4 polymorphism (HVR4), which is the result of a variable number of tandem repeats in intron 7; and (3) the RQ353 polymorphism, a guanine-to-adenine substitution in the position of the codon for amino acid 353 resulting in an amino acid replacement of arginine (R) by glutamine (Q) in the FVII protein. The frequencies of these three polymorphisms and their linkage disequilibrium were different in the four populations studied. The frequencies of the alleles associated with higher plasma FVII:C were lower in the Europeans than in the Inuit, a population with a lower incidence of IHD. There was an association between both the promoter polymorphism and the RQ353 polymorphism and the plasma FVII:C in the Europeans, the Inuit, and the Gujarati Indians, and an association only between the RQ353 polymorphism and plasma FVII:C in the Afrocaribbeans. Only in the Inuit was the HVR4 polymorphism associated with plasma FVII:C. In multiple regression analysis, the additional information provided by the promoter polymorphism when the other polymorphisms were already included in the model was the most pronounced, suggesting that the promoter polymorphism may be the functional mutation having the greatest effect on determining plasma FVII:C. PMID- 9351353 TI - Lovastatin decreases de novo cholesterol synthesis and LDL Apo B-100 production rates in combined-hyperlipidemic males. AB - The effect of lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity, on the kinetics of de novo cholesterol synthesis and apolipoprotein (apo) B in very-low-density lipoprotein (VLDL), intermediate density lipoprotein (IDL), and low-density lipoprotein (LDL) was investigated in five male patients with combined hyperlipidemia. Subjects were counseled to follow a Step 2 diet and were treated with lovastatin and placebo in randomly assigned order for 6-week periods. At the end of each experimental period, subjects were given deuterium oxide orally and de novo cholesterol synthesis was assessed from deuterium incorporation into cholesterol and expressed as fractional synthesis rate (C-FSR) and production rate (C-PR). Simultaneously, the kinetics of VLDL, IDL, and LDL apo B-100 were studied in the fed state using a primed-constant infusion of deuterated leucine to measure fractional catabolic rates (FCR) and production rates (PR). Drug treatment resulted in significant decreases in total cholesterol (-29%), VLDL cholesterol (-40%), LDL cholesterol ( 27%), and apo B (-16%) levels and increases in HDL cholesterol (+13%) and apolipoprotein (apo) A-I (+11%) levels. Associated with these plasma lipoprotein responses was a significant reduction in both de novo C-FSR (-40%; P = .04) and C PR (-42%; P = .03). Treatment with lovastain in these patients had no significant effect on the FCR of apoB-100 in VLDL, IDL, or LDL, but resulted in a significant decrease in the PR of apoB-100 in IDL and LDL. Comparing the kinetic data of these patients with those of 10 normolipidemic control subjects indicates that lovastatin treatment normalized apoB-100 IDL and LDL PR. The results of these studies suggest that the declines in plasma lipid levels observed after treatment of combined hyperlipidemic patients with lovastatin are attributable to reductions in the C-FSR and C-PR of de novo cholesterol synthesis and the PR of apoB-100 containing lipoproteins. The decline in de novo cholesterol synthesis, rather than an increase in direct uptake of VLDL and IDL, may have contributed to the decline in the PR observed. PMID- 9351352 TI - Dysregulation of monocytic nuclear factor-kappa B by oxidized low-density lipoprotein. AB - Nuclear factor-kappa B (NF-kappa B)/Rel transcription factors may be involved in atherosclerosis, as is suggested by the presence of activated NF-kappa B in human atherosclerotic lesions. The aim of the present study was to investigate the effects of oxidized LDL (oxLDL) on the NF-kappa B system in human THP-1 monocytic cells as well as adherent monocytes. Our results demonstrate that short-term incubation of these cells with oxLDL activated p50/p65 containing NF-kappa B dimers and induced the expression of the target gene IL-8. This activation of NF kappa B was inhibited by the antioxidant and H2O2 scavenger pyrrolidine dithiocarbamate and the proteasome inhibitor PSI. The oxLDL-induced NF-kappa B activation was accompanied by an initial depletion of I kappa B-alpha followed by a slight transient increase in the level of this inhibitor protein. In contrast, long-term treatment with oxLDL prevented the lipopolysaccharide-induced depletion of I kappa B-alpha, accompanied by an inhibition of both NF-kappa B activation and the expression of tumor necrosis factor-alpha and interleukin-1 beta genes. These observations provide additional evidence that oxLDL is a potent modulator of gene expression and suggest that (dys)regulation of NF-kappa B/Rel is likely to play an important role in atherogenesis. PMID- 9351355 TI - Possible mechanisms of collar-induced intimal thickening. AB - The positioning of a soft silicone collar around the rabbit carotid artery induces intimal thickening. We investigated to which extent occlusion of the vasa vasorum, damage of the perivascular nerve network, and/or changes in blood flow velocity contribute to intimal thickening. To this end, collars with different bores (diameter of inlet and outlet) were positioned around the carotid artery of male rabbits for 14 days. In another experiment, 75% of the wall of fitting collars was removed (open collar). In the midcollar region, the cross-sectional area of the intima reached a maximum (72 +/- 14 mm2/1000) when the endings of the collar fitted the artery closely. Removal of the side wall of these fitting collars reduced intimal thickening by 90%. Examination of unoperated carotid arteries never showed penetration of the adventitia or the media by vasa vasorum. The perivascular neuronal network in the region surrounded by a closed or an open collar was almost completely lost as compared with the zones outside the collar. Both the closed and open collar slightly bent the artery and increased the peak systolic velocity, measured with pulsed color Doppler after 6 hours, to a similar extent as compared with the proximal zone outside the collar. After 2 weeks, the peak systolic velocity within both the closed and open collar was partly normalized and was statistically not different from the proximal zone outside the collar. In conclusion, the geometry of the collar influenced the extent of intimal thickening, whereby more intimal thickening was obtained with a collar whose endings fit the carotid artery, rather than with a loose collar. Moreover, a closed structure was essential. The results obtained with the open collar exclude occlusion of vasa vasorum, damage of the perivascular neuronal network, kinking of the artery, and changes in blood flow velocity as major factors in the collar-induced intimal thickening. Our findings are consistent with the possibility that intimal thickening is the consequence of the combination of both vascular injury and hindrance of transmural flow by the collar. The obstruction of transmural fluid transport may then lead to retention of toxic metabolites, and/or cytokines within the segment enclosed by the collar. PMID- 9351356 TI - Thrombin receptor-mediated increase of two matrix metalloproteinases, MMP-1 and MMP-3, in human endothelial cells. AB - Matrix metalloproteinases (MMPs) are responsible for the degradation of extracellular matrix components and are secreted by a variety of cells including human endothelial cells. Because alpha-thrombin is known to interact with matrix components and has been shown to activate latent MMP-2 in human umbilical vein endothelial cells, we investigated whether human alpha-thrombin could also regulate other MMPs secreted by the human saphenous vein or mammary artery endothelial cells (EC). After treatment of EC with increasing concentrations of thrombin for different periods of time, a significantly higher gelatinolytic activity of both MMP-1 and MMP-3 was observed in addition to MMP-2 activation. The effect of thrombin was time and dose-dependent, reaching a maximum at 24 hours. After treatment with 5 NIH U/ml thrombin for 24 hours, Western blotting revealed 9.5- and 4.4-fold increases over control values for MMP-3 and MMP-1, respectively. The synthetic thrombin receptor agonist peptide SFLLRNPNDKYEPF fully reproduced the action of thrombin, whereas chemical inactivation of the catalytic site of thrombin abolished its effect on MMP-1 and MMP-3. Thrombin and SFLLRNPNDKYEPF both induced MMP-3 mRNA synthesis but had no significant influence on constitutive MMP-1 mRNA levels. These results demonstrate that thrombin not only activates latent MMP-2 but also modulates MMP-1 and MMP-3 production in EC, this latter effect being mediated by the G-protein-coupled thrombin receptor. Hence, our present data provide evidence to support the suspected role of thrombin in tissue remodeling and angiogenesis. PMID- 9351357 TI - Folic acid deficiency enhances oral contraceptive-induced platelet hyperactivity. AB - In previous studies conducted in female rats and in women, oral contraceptives (OC) were found to induce a platelet hyperactivity that was related to an oxidative stress. Because cases of megaloblastic anemia have been reported to occur in women taking OC, these treatments are suspected of depleting folate stores. In the study presented herein, which was conducted in rats, we sought to determine the influence of dietary folic acid deficiency (FD) on the thrombogenicity of OC. Animals were fed for 6 weeks with either a folic acid deficient diet (250 micrograms/kg folic acid) or a control diet (750 micrograms/kg). One-half of the animals in each group were treated with OC (ethinyl estradiol plus lynestrenol). FD and OC individually potentiated platelet aggregation in response to thrombin and ADP and the release and metabolism of arachidonic acid, in particular, the biosynthesis of thromboxane. These platelet activities were further enhanced in animals given both the folic acid-deficient diet and the OC treatment. In addition, FD enhanced the pro-oxidant state in OC treated rats characterized by (1) a fall in platelet and plasma n-3 fatty acids, (2) an increase in plasma lipid peroxidation products such as conjugated dienes, lipid peroxides, and thiobarbituric reactive substances, (3) a rise in ex vivo erythrocyte susceptibility to free radicals. Moreover, we found that OC treatment led to a reduction of plasma and erythrocyte folate concentrations associated with a moderate hyperhomocysteinemia. Under our experimental conditions, we did not find significant synergistic effects between OC and FD. We propose that, although the untoward effects associated with the OC treatment may not primarily be dependent on FD, the folic acid deficiency magnified OC-induced oxidative stress, which resulted in platelet hyperactivity by elevating the pro-oxidant homocysteine plasma concentration. Despite the limitations of this animal model, the data of the present study suggest that in addition to cigarette smoking, inadequate folic acid intake might predispose those taking OC to vascular thrombosis. PMID- 9351358 TI - Homocyst(e)ine and risk of cardiovascular disease in the Multiple Risk Factor Intervention Trial. AB - A nested case-control study was undertaken involving men participating in the Multiple Risk Factor Intervention Trial (MRFIT). Serum samples from 712 men, stored for up to 20 years, were analyzed for homocyst(e)ine. Cases involved nonfatal myocardial infarctions (MIs), identified through the active phase of the study, which ended on February 28, 1982, and deaths due to coronary heart disease (CHD), monitored through 1990. The nonfatal MIs occurred within 7 years of sample collection, whereas the majority of CHD deaths occurred more than 11 years after sample collection. Mean homocyst(e)ine concentrations were in the expected range and did not differ significantly between case patients and control subjects: MI cases, 12.6 mumol/L; MI controls, 13.1 mumol/L; CHD death cases, 12.8 mumol/L; and CHD controls, 12.7 mumol/L. Odds ratios versus quartile 1 for CHD deaths and MIs combined were as follows: quartile 2, 1.03; quartile 3, 0.84; and quartile 4, 0.92. Thus, in this prospective study, no association of homocyst(e)ine concentration with heart disease was detected. Homocyst(e)ine levels were weakly associated with the acute-phase protein (C-reactive protein). These results are discussed with respect to the suggestion that homocyst(e)ine is an independent risk factor for heart disease. PMID- 9351359 TI - Cytokine modulation of LDL oxidation by activated human monocytes. AB - There is considerable evidence to suggest that cytokines modulate the pathological cellular events that occur in human atherosclerosis. We sought to determine the effects of T-helper-lymphocyte (TH)-1- and TH2-type cytokines on the ability of human monocytes to oxidize LDL, one of the pathological processes believed to occur in atherosclerosis. The ability of opsonized zymosan (ZOP) activated human monocytes to oxidize LDL in a 24-hour period was significantly enhanced by pretreatment of the monocytes with the TH2 cytokines, interleukin (IL)-4, or IL-13 compared with untreated monocytes. In contrast, interferon (IFN) gamma, a TH1 cytokine, inhibited LDL oxidation by activated monocytes. Treatment with IFN-gamma also prevented the IL-4- and IL-13-mediated enhancement of LDL oxidation by ZOP-activated monocytes. Untreated or cytokine-treated unactivated monocytes did not oxidize LDL. The enhancement of LDL oxidation mediated by IL-4 or IL-13 treatment was not due to a mitogenic effect of the cytokines on the monocytes, nor to modulation of superoxide anion (O2-) production. The cytokine regulation of 15-lipoxygenase (LO) in the monocytes was also examined. IL-4 and IL-13 induction of 15-LO mRNA and 15-LO activity in the monocytes was confirmed, as was the previously reported inhibition of induction by IFN-gamma. In summary, IL-4 and IL-13 enhance the ability of activated human monocytes to oxidize LDL, whereas IFN-gamma inhibits the cell-mediated oxidation. The up- and downregulation of activated monocyte-mediated LDL oxidation by these cytokines correlates with the expression of 15-LO activity. Considerable evidence suggests that the progression of atherosclerosis includes events that are immunologically mediated, lending potential physiological relevance to these in vitro observations. PMID- 9351360 TI - High concentration of glucose increases mitogenic responsiveness to heparin binding epidermal growth factor-like growth factor in rat vascular smooth muscle cells. AB - The effect of a high extracellular glucose concentration on the mitogenic response of rat vascular smooth muscle cells (SMCs) to heparin-binding epidermal growth factor-like growth factor (HB-EGF) was investigated. The mitogenic effect of HB-EGF was significantly greater in SMCs cultured in high glucose (25 mmol/L) than in cells cultured in low glucose (5.5 mmol/L) or at high osmolarity (5.5 mmol/L glucose plus 19.5 mmol/L mannitol). The mitogenic effect of epidermal growth factor (EGF), which shares the EGF receptor with HB-EGF, was not affected by glucose concentration. The mitogenic effect of HB-EGF was greater when incubated with heparan sulfate (HS) isolated from SMCs cultured in high glucose than with HS from cells cultured in low glucose. HS synthesized by cells in high glucose was of smaller molecular size and less sulfated than HS synthesized by cells in low glucose. The abundance of mRNA encoding HS-N-deacetylase/N sulfotransferase (HS-NdAc/NST), a regulatory enzyme in the biosynthesis of HS, was decreased by high glucose in a protein kinase C-independent manner. These observations suggest that the enhanced mitogenic response to HB-EGF in SMCs cultured in high glucose may be attributable to changes in cell-associated HS. Downregulation of HS-NdAc/NST gene expression by high glucose may be related to the altered HS biosynthesis. PMID- 9351361 TI - A common mutation in the lipoprotein lipase gene promoter, -93T/G, is associated with lower plasma triglyceride levels and increased promoter activity in vitro. AB - Single-strand conformational polymorphism analysis of the lipoprotein lipase promoter identified a T-->G transition at position -93. The frequency in healthy white men was 3.4% (n = 1575). There was an 83% allelic association between -93T- >G and Asp9-->Asn (D9N); all N9 mutations occurred on a -93G allele, but not all 93G mutations occurred on an N9 allele. It was thus possible to assess the effect on plasma triglyceride (Tg) levels of the rare -93G mutation in the presence of the wild-type D9. Carriers of the -93G, with genotype TG/DD, had significantly lower Tg levels than TT/DD individuals (1.36 versus 1.78 mmol/L, P = .01); carriers of both mutations (TG/DN) had the highest Tg levels (1.93 mmol/L). When the group was stratified above and below the sample mean for body mass index (BMI), carriers of the -93G on a D9 allele (TG/DD) were "protected" against the Tg-raising effect of obesity, as assessed by BMI. In Afro-Caribbeans (n = 91), the carrier frequency of -93G was 18-fold higher (63%), with weaker (17%) allelic association between -93G and N9. In vitro, the -93G promoter had 24% higher activity than the -93T in a rat smooth muscle cell line and 18% higher activity in a human adrenal cell line. A protein identified by band-shift assays bound to the -93G but not to the -93T allele, which may explain the lower Tg levels in 93G carriers. PMID- 9351362 TI - Role of tyrosine kinases in extracellular matrix-mediated modulation of arterial smooth muscle cell phenotype. AB - Fibronectin (FN) promotes the modulation of freshly isolated arterial smooth muscle cells (SMCs) from a contractile to a synthetic phenotype by interacting with integrins on the cell surface. This process is characterized by a structural and functional transformation of the cells, including a reorganization of the cytoskeleton, the formation of a large secretory apparatus, and the acquisition of proliferative capacity. In this study we have investigated the role of integrin signaling through tyrosine kinases in the structural changes that occur in SMCs during primary culture on FN. A gradual increase in phosphotyrosine staining in focal adhesions and a concomitant increase in tyrosine phosphorylation of proteins including focal adhesion kinase were observed. In contrast, cells seeded on laminin formed few focal adhesions, and tyrosine phosphorylation of proteins was less than in cells cultured on FN. Treatment of cells cultured on FN with the tyrosine kinase inhibitor genistein strongly suppressed focal adhesion formation, cell spreading, and cytoskeletal reorganization. In addition, electron microscopic analysis demonstrated that the phenotypic modulation was slowed down. These results indicate that the ability of extracellular matrix components to promote a change in the phenotypic properties of SMCs depends on the assembly of focal adhesions with associated tyrosine kinase activity. PMID- 9351363 TI - Effects of angiotensin II on cardiac function and peripheral vascular structure during compensated heart failure in the rat. AB - The present experiments were designed to test the hypothesis that the activation of the renin-angiotensin system during compensated heart failure may have adverse effects on cardiac function and change the peripheral vascular structure. ANG II (250 ng/kg/min) or saline (0.9% NaCl) were infused in myocardial-infarcted and sham-operated rats. After 2 weeks, cardiac function and peripheral vascular changes were investigated. RESULTS: ANG II infusion reduced baseline cardiac index in sham rats but did not further reduce this index in ANG II-infused MI rats. Total peripheral resistance was similarly increased in ANG II-infused infarcted and sham rats, and also plasma ANG II concentrations were comparable. ANG II elevated systolic blood pressure by approximately 70 mm Hg in sham rats and increased the medial cross-sectional area of the superior mesenteric artery by 33%. However, ANG II infusions in MI rats resulted in only a minor increase in blood pressure, whereas the cross-sectional area of the superior mesenteric artery did not change. ANG II infusion had no effect on vessel dimensions of the resistance arteries of the pulmonary and mesenteric vascular bed of either group. Calculated ED50 and peak pressor response to acute ANG II injections were comparable in all groups, confirming the presence of functionally intact AT1 receptors. The increases in plasma atrial natriuretic peptide (ANP) and nitric oxide (NO) synthase activity (estimated by aortic cyclic GMP concentrations) were higher in ANG II-infused MI rats than in ANG II-infused sham rats. CONCLUSION: ANG II infusion in rats with and without MI has comparable negative effects on cardiac function but has different effects on blood pressure and vascular structure. The concomitant increases in plasma ANP and NO synthase activity in ANG II-infused MI rats suggest that the growth stimulatory and hypertensive actions of ANG II in sham rats may be counter-regulated by activation of inhibitory neurohumoral systems such as ANP or NO in MI rats. PMID- 9351364 TI - Effect of bone marrow transplantation on lipoprotein metabolism and atherosclerosis in LDL receptor-knockout mice. AB - The LDL receptor (LDLR) plays an important role in the removal of LDL and its precursors, the intermediate and very low density lipoproteins, from the blood circulation. The receptor is expressed on various cell types. In this study the relative importance of the LDLR on macrophages for lipoprotein metabolism and atherogenesis was assessed. For this purpose, irradiated LDLR-knockout (-/-) mice were transplanted with bone marrow of normal C57BL/6J mice. DNA analysis showed that the transplanted mice were chimeric. The transplantation resulted in a slight decrease of total serum cholesterol when compared with LDLR-/- mice that were transplanted with LDLR-/- bone marrow. This modest decrease, however, did not reach statistical significance at all time points examined. This decrease can be almost completely attributed to a decrease in LDL cholesterol. The specific lowering of LDL cholesterol could clearly be observed at 4 weeks after transplantation, but the decrease was less at 12 weeks after transplantation. Quantification of atherosclerotic lesions of mice fed a 1% cholesterol diet for 6 months revealed that there were no differences in mean lesion area between mice transplanted with wild-type bone marrow or LDLR-/- bone marrow. We anticipate that in LDLR-/- mice transplanted with wild-type bone marrow, the LDLR is downregulated by the relatively high concentrations of circulating cholesterol. In vitro incubations of peritoneal macrophages with 125I-LDL indicated that the LDLR of these cells could be downregulated by 25-hydroxycholesterol. Peritoneal macrophages isolated from LDLR-/- mice transplanted with wild-type bone marrow, in contrast to those transplanted with LDLR-/- bone marrow, were able to degrade 125I-LDL, indicating that the capacity to express functional LDLR was achieved. In conclusion, introduction of the LDLR into LDLR -/- mice via bone marrow transplantation resulted in only a relatively modest decrease of LDL cholesterol that became less pronounced at later time points, possibly due to downregulation of the LDLR. To utilize the LDLR in macrophages for effective cholesterol lowering, either the sterol-regulatory elements have to be "silenced" or a high expression LDLR construct has to be introduced into macrophages, eg, via transplantation of in vitro transfected hematopoietic stem cells. PMID- 9351365 TI - Androgen deprivation is associated with enhanced endothelium-dependent dilatation in adult men. AB - Male gender is an independent risk factor for coronary artery disease, and androgen administration has been associated with increased atherosclerosis in experimental animals. Since endothelial dysfunction is an important event in the atherogenic process, we hypothesized that androgen deprivation in adult men might be associated with enhanced arterial endothelial function. Using external vascular ultrasound, brachial artery diameter was measured at rest, after flow increase (causing endothelium-dependent dilatation) and after nitroglycerin (an endothelium-independent dilator). We studied 30 adult males aged 40 to 70 years: 10 had had bilateral orchidectomy and/or maximal androgen blockade for > or = 6 months for treatment of prostate cancer, and all were in complete remission (group 1). Ten healthy controls (group 2) and 10 controls who had remission from nonprostate cancers (group 3) were matched for age and smoking history. Testosterone levels were lower in men in group 1 versus groups 2 or 3 (0.8 +/- 0.1 versus 19.2 +/- 8.4 or 16.1 +/- 4.9 nmol/L, P < .001). By contrast, endothelium-dependent dilatation was markedly higher in group 1 than in groups 2 or 3 (6.2 +/- 3 versus 2.7 +/- 2 or 2.0 +/- 1.9%, P < .001). The nitroglycerin response was similar in all three groups (P = .92). On multivariate analysis, increased endothelium-dependent dilatation was significantly associated with low serum testosterone levels (P = .001) but not with cholesterol levels or with a past history of malignancy (P > .25). The withdrawal of male sex hormones may be associated with enhanced endothelial function in adult men. This is consistent with a deleterious effect of physiologic levels of male sex steroids on the arterial wall. PMID- 9351366 TI - Effects of genotype and diet on cholesterol efflux into plasma and lipoproteins of normal, apolipoprotein A-I-, and apolipoprotein E-deficient mice. AB - We investigated the contribution of apoE to cholesterol efflux into plasmas of normal, apoA-I-, and apoE-deficient mice, which were fed with chow- and cholesterol-rich diets. Plasmas of normal and apoA-I-deficient mice contain apoE in pre-beta-migrating VLDL as well as in HDL-like lipoproteins, which have either electrophoretic alpha- or gamma-mobilities. The latter particle resembled gamma LpE in human plasma also by its mobility on nondenaturing two-dimensional electrophoresis. No apoE-containing lipoproteins were found in plasmas of apoE deficient mice. When apoA-I- and apoE-deficient mice received both chow- and fat rich diets, their plasmas released significantly less 3H-cholesterol from radiolabeled fibroblasts than did plasma of normal mice. Removal of apoE from plasmas of normal and apoA-I-deficient mice by anti-apoE immunoaffinity chromatography decreased their cholesterol efflux capacities (per 1 minute/per 1 hour) by 26%/40% (P = 0.0092/0.0007) and 30%/26% (P = 0.0092/0.0003), respectively. Net cholesterol efflux from fibroblasts into apoA-I-deficient plasma was 45% lower compared with plasma of normal mice. Incubation of fibroblasts with apoE-deficient plasma caused net influx of cholesterol. Prior addition of human apoE to or removal of apoB-containing lipoproteins from apoE deficient plasma restored its ability to cause net cholesterol efflux to 50% of normal plasma. Some of the differences between cholesterol efflux into normal and apoE-deficient plasmas were attributable to the failure of apoE-deficient plasmas to take up cell-derived 3H-cholesterol into gamma-LpE. Compared with normal plasma, both apoA-I-deficient and apoE-deficient plasmas were significantly decreased in their activity to esterify cell-derived 3H-cholesterol. Anti-apoE chromatography decreased significantly cholesterol esterification in normal plasma and apoA-I-deficient plasma but not in apoE-deficient plasma. Taken together, the data provide evidence that apoE is an important contributor to reverse cholesterol transport, partially because of initial uptake of cell derived cholesterol by gamma-LpE and partially because of the contribution of apoE-containing lipoproteins to esterification of cholesterol in plasma. PMID- 9351367 TI - Niacin decreases removal of high-density lipoprotein apolipoprotein A-I but not cholesterol ester by Hep G2 cells. Implication for reverse cholesterol transport. AB - Niacin (nicotinic acid) is the most potent clinically used agent for increasing plasma HDL and apolipoprotein (apo) A-I. The mechanism by which niacin increases apoA-I is not clearly understood. We have examined the effect of niacin on the hepatic production and removal of apoA-I using Hep G2 cells as an in vitro model. Incubation of Hep G2 cells with niacin resulted in increased accumulation of apoA I in the medium in a dose-dependent manner. Incorporation of [3H]leucine and [35S]methionine into apoA-I and apoA-I mRNA expression was unchanged by niacin, suggesting that it did not affect apoA-I de novo synthesis. Uptake of radiolabeled HDL protein and HDL apoA-I by Hep G2 cells was significantly reduced to as much as 82.9 +/- 2.2% (P = .04) and 84.2 +/- 2.8% (P = .02), respectively, of the baseline with increasing concentrations of niacin (0 to 3.0 mmol/L). Specific 125I-HDL protein uptake measured with a 50-fold excess of unlabeled HDL was reduced to as much as 78.3 +/- 4.8% (P = .005) in niacin-treated cells. The uptake of labeled cholesterol esters in HDL was unaffected by niacin. Niacin also effected a similar decrease in HDL protein uptake, but not cholesterol esters, from apoA-I-containing HDL particles isolated by immunoaffinity. The conditioned medium obtained from Hep G2 cells incubated with niacin significantly (P = .002) increased cholesterol efflux from cultured human fibroblasts. These data indicate a novel mechanism whereby niacin selectively decreases hepatic removal of HDL apoA-I but not cholesterol esters, thereby increasing the capacity of retained apoA-I to augment reverse cholesterol transport. PMID- 9351368 TI - Response of vascular smooth muscle cells to the neuropeptide secretoneurin. A functional role for migration and proliferation in vitro. AB - Mesenchymal cell migration and replication are central biologic events involved in atherosclerosis and lung and hepatic fibrosis. Tissue repair and fibrosis are thought to be regulated by growth regulatory molecules, comprising both stimulators and inhibitors of mesenchymal cell functions, including platelet derived growth factor (PDGF), transforming growth factor-beta (TGF-beta), fibroblast growth factors, and several neuropeptides such as substance P. Secretoneurin (SN), a novel 33-amino acid neuropeptide derived from secretogranin II (chromogranin C), is widely distributed in the central and peripheral nervous and neuroendocrine systems, including afferent C-fibers, and can be released in the periphery by capsaicin. Recently, we reported that SN triggers the selective migration of human monocytes and fibroblasts in vitro, implicating its involvement in inflammatory responses. We report herein that SN stimulates specific migration (maximal response at 10(-10) M) of cultured arterial smooth muscle cells (SMCs), originating from rat thoracic aorta, and initiates DNA synthesis and SMC growth (BrdU incorporation, MTT test) with a maximum at 10(-8) M SN to a similar extent as observed by PDGF. Both functional activities of SN were inhibited by specific anti-SN immunoglobulins (dilution, 1:1000), and furthermore, a trypsinized SN peptide (10(-8) M) was unable to provoke biologic effects. Our studies suggest that SN functions as a regulatory peptide to modulate SMC migration and proliferation, which in conjunction with other factors could serve to aggravate and accelerate the development of atherosclerotic or restenotic lesions at sites of vascular injury. PMID- 9351369 TI - Lipoprotein(a) isoforms display differences in affinity for plasminogen-like binding to human mononuclear cells. AB - Binding of lipoprotein(a) (Lp(a)) to membrane proteins of the monocyte-macrophage cell lineage may be an important event in atheroma formation. Since Lp(a) with distinct apolipoprotein(a) (apo(a)) isoforms may show differences in their affinity with regard to fibrin binding, the existence of such a functional behavior in the interaction of apo(a) in Lp(a) with these cells was explored using the monocytic cell line THP-1. Lp(a) preparations containing small size apo(a) isoforms (M(r) = 450,000 to 550,000) and high molecular mass isoforms (M(r) > or = 700,000) were purified from plasmas containing > 0.35 g/L of Lp(a) obtained from subjects (n = 14) with cardiovascular atherosclerotic disease. Binding of plasminogen to THP-1 cells was performed using the method of radioisotopic dilution. For binding of Lp(a) to cells, the THP-1 monocytic cells were incubated with varying concentrations of the different Lp(a) preparations; cells were then washed and the amount of Lp(a) bound was detected with a radiolabeled polyclonal antibody directed against apo(a). Binding due to kringle interactions with lysine residues was calculated by subtracting from the total bound the amount of Lp(a) bound (approximately 10%) in the presence of 6 aminohexanoic acid. Analysis of data with the Langmuir equation indicated identical and independent (non-interacting) sites and allowed evaluation of the Kd. Binding isotherms of small size isoforms showed saturation and a high affinity (Kd = 25.8 +/- 19 nmol/L) relative to that of plasminogen (Kd = 1750 +/- 760 nmol/L). A similar difference (Kd = 17.5 +/- 7.9 nmol/L versus Kd = 600 +/- 220 nmol/L) was found when binding experiments were performed with a fibrin surface. In contrast, binding isotherms of the high molecular mass isoforms did not show saturation at the highest Lp(a) concentrations used, thus indicating a lower affinity. In conclusion, these results show that apo(a) isoforms may display polymorphism-linked functional heterogeneity with regard to cell binding, which may explain the higher association with cardiovascular risk of small size isoforms. These qualitative differences in the binding of apo(a) isoforms to fibrin or cells may modulate the cardiovascular risk associated with high levels of Lp(a). PMID- 9351370 TI - Altered platelet function detected by flow cytometry. Effects of coronary artery disease and age. AB - Platelet activation state and responsiveness to physiological agonists were measured in 65 patients with documented coronary artery disease (54 male and 11 female; mean age, 58 years). Twelve patients (mean age, 52 years), selected at random from the male cohort, were compared with 12 age-matched male control subjects (mean age, 52 years) and with 10 normal, young male subjects (mean age, 25 years). Whole-blood flow cytometry was used to measure platelet activation status ex vivo and platelet responsiveness to physiological agonists in vitro. Peripheral blood samples were analyzed for bound fibrinogen and expression of P selectin, GPIb, and GPIIb-IIIa at rest and in response to ADP (0.1 to 10 mumol/L) and thrombin (0.02 to 0.32 mu/mL). No significant differences were seen in the basal levels of fibrinogen binding between any of the groups, but P-selectin expression was significantly lower in patients compared with age-matched control subjects (P = .0005). When stimulated with agonists, patients' platelets had significantly decreased fibrinogen binding (P < .03) but no difference in P selectin expression compared with the age-matched group. Both agonist-induced fibrinogen binding and P-selectin expression were, however, higher in the young subjects compared with either the older control group or the patients (P < .05). GPIb and GPIIb-IIIa expression were lowest in the patients with angina and highest in the young control subjects, with levels in the age-matched control subjects falling between these values. Data from the total patient cohort (n = 65) were identical to those in the smaller cohort (n = 12). In conclusion, atherosclerosis impairs platelet aggregatory responses (fibrinogen binding) over and above the decreased response seen with age. Platelet degranulation (P selectin expression) is also impaired in patients with coronary artery disease, but only in comparison with younger subjects, not age-matched controls. PMID- 9351372 TI - Evaluation of endothelial shear stress and 3D geometry as factors determining the development of atherosclerosis and remodeling in human coronary arteries in vivo. Combining 3D reconstruction from angiography and IVUS (ANGUS) with computational fluid dynamics. AB - The predilection sites of atherosclerotic plaques implicate rheologic factors like shear stress underlying the genesis of atherosclerosis. Presently no technique is available that enables one to provide 3D shear stress data in human coronary arteries in vivo. In this study, we describe a novel technique that uses a recently developed 3D reconstruction technique to calculate shear stress on the endothelium with computational fluid dynamics. In addition, we calculated local wall thickness, the principal plane of curvature, and the location of plaque with reference to this plane, relating these results to shear stress in a human right coronary artery in vivo. Wall thickness and shear stress values for the entire vessel for three inflow-velocity values (10 cm/second, 20 cm/second, and 30 cm/second equivalents with the Reynolds numbers 114,229, and 457) were as follows: 0.65 +/- 0.37 mm (n = 1600) and 19.6 +/- 1.7 dyne/cm2; 46.1 +/- 8.1 dyne/cm2 and 80.1 +/- 16.8 dyne/cm2 (n = 1600). Curvature was 25 +/- 9 (m-1), resulting in Dean numbers 20 +/- 8; 46 +/- 16, and 93 +/- 33. Selection of data at the inner curvature of the right coronary artery provided wall thickness values of 0.90 +/- 0.41 mm (n = 100), and shear stress was 17 +/- 17, 38 +/- 44, and 77 +/- 54 dyne/cm2 (n = 100), whereas wall thickness values at the outer curve were 0.37 +/- 0.17 mm (n = 100) and shear stress values were 22 +/- 17, 60 +/- 44, and 107 +/- 79 dyne/cm2 (n = 100). These findings could be reconciled by an inverse relationship between wall thickness and shear stress for each velocity level under study. For the first time for human vessels in vivo, evidence is presented that low shear stress promotes atherosclerosis. As the method is nondestructive, it allows repeated measurements in the same patient and will provide new insights in the progress of atherosclerosis. PMID- 9351371 TI - Heritability analysis of lipids and three gene loci in twins link the macrophage scavenger receptor to HDL cholesterol concentrations. AB - We studied 100 healthy monozygotic and 72 dizygotic twin pairs (mean age, 34 +/- 14 years) to test for genetic influences on blood lipids and to examine relevant gene loci. Total cholesterol (TC), LDL cholesterol (LDL-C), HDL cholesterol (HDL C), and triglyceride (TG) levels were determined after a 12-hour fast. Zygosity was determined with the use of microsatellite markers. Heritability estimates were conducted by using the lisrel 8 program; a sib-pair analysis was conducted by using the sibpal program. Linear regression analyses were carried out between identical-by-descent status and squared within-pair differences of TC, LDL-C, HDL C, and TG values. Heritability estimates of the lipid serum concentrations ranged from .58 to .66. A significant linkage relationship was found for HDL-C (P = .008) and TGs (P = .05) with D8S261 on chromosome 8p. However, no linkage was found between any of the lipid variables and the lipoprotein lipase gene locus (LPL GZ14/15 and D8S282). Because D8S261 is located approximately halfway between the LPL and macrophage scavenger receptor genes, we examined the nearby markers D8S549 and D8S1731. Linkage was found for HDL-C and D8S549 (P = .001) and for HDL C and D8S1731 (P = .04). On the other hand, we found no linkage between the LDL receptor gene locus and LDL-C serum concentrations nor between the LPL gene locus and the various other lipid fractions. Our data suggest a significant influence of the macrophage scavenger receptor gene locus on HDL-C and weak influence on TG levels. We suggest that inherited variability in the macrophage scavenger receptor gene has an influence on serum lipid concentrations. PMID- 9351373 TI - Modified LDL decreases the binding of prostaglandin E2, I2, and E1 onto monocytes in patients with peripheral vascular disease. AB - Recent data suggest that various eicosanoids including prostaglandins play an important regulatory role in the development of atherosclerotic lesions. Peripheral blood monocytes have been implemented in early atherogenesis because they express receptors specific for modified LDL. In this study we investigated the binding of tritium prostaglandins E2 (3H-PGE2), E1 (3H-PGE1) and I2 (3H-PGI2) onto intact peripheral monocytes isolated from 20 patients (32-71 years) with manifested ischemic peripheral vascular disease stage II according to Fontaine and compared the results with those obtained in 16 healthy volunteers (21-68 years). In control subjects, Scatchard analyses of the binding data indicated a single class of high-affinity binding sites for 3H-PGE2 (maximal binding capacity [Bmax] = 11,400 +/- 3200 sites/cell; dissociation constant [Kd] = 1.3 +/- 0.5 nmol/L) and two classes of binding sites for 3H-PGE1 (Bmax1 = 11,200 +/- 4900 sites/cell, Kd1 = 1.5 +/- 0.5 nmol/L; Bmax2 = 47,800 +/- 6100 sites/cell, Kd2 = 12.8 +/- 5.9 nmol/L) as well as for 3H-PGI2 (Bmax1 = 10,100 +/- 3700 sites/cell, Kd1 = 1.7 +/- 0.7 nmol/L; Bmax2 = 81,200 +/- 5200 sites/cell, Kd2 = 14.2 +/- 6.5 nmol/L). In the patients, an absence of the higher-affinity binding class and significantly (P < .01) fewer lower-affinity binding sites were found for each ligand (PGE2: Bmax = 6600 +/- 3600 sites/cell, Kd = 12.1 +/- 3.2 nmol/L; PGI2: Bmax = 6400 +/- 3100 sites/cell, Kd = 22.1 +/- 8.3; PGE1: Bmax = 5300 +/- 1700 sites/ cell, Kd = 20.5 +/- 7.0 nmol/L). After incubation of monocytes with modified LDL (oxidized LDL or acetylated LDL), the binding of prostaglandins was significantly (P < .01 to P < .001) decreased, whereas native VLDL, LDL, and HDL did not interfere with prostaglandin binding. Prostaglandin-induced adenosine 3' 5' cyclic monophosphate (cAMP) formation by monocytes was significantly (P < .01) lower in patients (the concentrations causing 50% elevation of basal cAMP formation [ED50] were 3.8 +/- 2.4 nmol/L for PGE2, 6.3 +/- 3.5 nmol/L for PGE1, and 5.6 +/- 4.1 nmol/L for PGI2) than in the control subjects (ED50 was 1.6 +/- 1.2 nmol/L for PGE2, 4.8 +/- 2.5 nmol/L for PGE1, and 3.1 +/- 1.4 nmol/L for PGI2). After preincubation with modified LDL, the PG-induced cAMP production by monocytes was remarkably decreased in both patients and control subjects (P < .05). Our results suggest a direct effect of modified LDL on PGE2, PGE1, and PGI2 binding onto monocytes by reducing the number of cell surface-expressed receptors available. Modified LDL also reduces the sensitivity of monocytes to prostaglandins, which results in decreased cAMP production. The complex interactions between prostaglandins and lipoproteins may play an important role during atherogenesis. PMID- 9351374 TI - Homocysteine as a risk factor for vascular disease. Enhanced collagen production and accumulation by smooth muscle cells. AB - An increased plasma homocysteine level is an independent risk factor for vascular disease. However, the pathological mechanisms by which homocysteine promotes atherosclerosis are not yet clearly defined. Arterial smooth muscle cells cultured in the presence of homocysteine grew to a higher density and produced and accumulated collagen at levels significantly above control values. Homocysteine concentrations as low as 50 mumol/L significantly increased both cell density and collagen production. Cell density increased by as much as 43% in homocysteine-treated cultures. Homocysteine increased collagen production in a dose-dependent manner. Smooth muscle cells treated with homocysteine at concentrations observed in patients with hyperhomocysteinemia had collagen synthesis rates as high as 214% of control values. Likewise, collagen accumulation in the cell layer was nearly doubled in homocysteine-treated cultures. Addition of aquacobalamin to homocysteine-treated cultures controlled the increase in smooth muscle cell proliferation and collagen production. These results indicate a cellular mechanism for the atherogenicity of homocysteine and provide insight into a potential preventive treatment. PMID- 9351375 TI - Plasminogen activator inhibitor-1 (PAI-1) antigen plasma levels in subjects attending a metabolic ward: relation to polymorphisms of PAI-1 and angiontensin converting enzyme (ACE) genes. AB - Plasminogen activator inhibitor 1 (PAI-1) is a determinant of vascular events. Subjects in metabolic wards are at high risk for these events. The renin angiotensin system modulates plasma PAI-1 levels. An insertion (4G)/deletion (5G) polymorphism is involved in the regulation of the circulating levels of PAI-1. We have evaluated the levels of plasma PAI-1 in 208 individuals from our metabolic ward and correlated these levels with the 4G/5G genotype as well as with a genotype (homozygosity for a deletion polymorphism, DD genotype) of the angiotensin-converting enzyme (ACE) gene. Homozygosity for the insertion genotype (5G/5G) was associated with PAI-1 levels lower than those associated with the deletion genotype (4G/4G) (26.2x/:1.6 versus 33.7x/:1.7 ng/mL, P = .036). Plasma PAI-1 levels appeared to depend on the genotype (P = .014) as much as on age (P = .044), t-PA (P = .0001), or triglyceride levels (P = .005). The association between triglycerides and PAI-1 was significant in subjects carrying the 4G/4G and the 4G/5G genotypes (P = .013 and .036, respectively) but not in those with the 5G/5G genotype. When stratified according to PAI-1 and ACE genotypes, individuals homozygous for both deletions (4G/4G-DD genotypes) exhibited higher plasma PAI-1 levels compared with those of individuals without such homozygosities. However, this difference did not reach statistical significance. We conclude that in a group of subjects from a metabolic ward, a 4G/5G polymorphism of the PAI-1 gene exerts effects on plasma PAI-1 antigen levels comparable to those of established determinants. The association between triglycerides and plasma PAI-1 levels is genotype dependent. A trend to a positive interaction between ACE DD and PAI-1 4G/4G in the regulation of circulating plasma PAI-1 levels is present in this setting. PMID- 9351376 TI - Monounsaturated and polyunsaturated n-6 fatty acid-enriched diets modify LDL oxidation and decrease human coronary smooth muscle cell DNA synthesis. AB - Proliferation of smooth muscle cells (SMCs) plays an important role in atherosclerotic lesion progression. The purpose of this investigation was to examine the effect of diets differing in fatty acid composition on human coronary SMC entry in the cell proliferation cycle. Twenty-four healthy men and women were placed on four consecutive diets lasting 5 weeks each: (1) saturated fatty acid (SFA)-rich diet with palm oil; (2) monounsaturated fatty acid (MUFA)-rich diet with olive oil; (3) polyunsaturated fatty acid (PUFA) n-6-rich diet with sunflower oil; and (4) PUFA n-3-rich diet (3.8 g/d). All diets supplied 35% of calories as fat. Compared with the SFA diet, all unsaturated diets reduced LDL cholesterol. Resistance of LDL to oxidative modification was significantly increased during the MUFA period (P < .05). Human coronary SMCs were cultured and induced by sera derived from the different groups. 3H-Thymidine incorporation into doubling DNA was significantly (P < .01) reduced during the MUFA and PUFA n 6 periods but not during the PUFA n-3 diet with respect to the SFA diet. This effect was more pronounced in women than in men. In conclusion, the MUFA-enriched diet reduced SMC DNA synthesis and LDL levels and protected LDL from oxidation. Therefore, these combined effects suggest that an oleic acid-rich Mediterranean diet could be better than PUFA (n-6)- or PUFA (n-3)-rich diets in the prevention of atherosclerosis. PMID- 9351377 TI - Postprandial elevation of ApoB-48-containing triglyceride-rich particles and retinyl esters in normolipemic males who smoke. AB - Smokers have an increased risk for coronary artery disease (CAD), which can only partly be explained by fasting lipoprotein changes. Recent studies have indicated that smokers express metabolic abnormalities characteristic of insulin resistance syndrome. A preliminary study reported an increased postprandial triglyceride (TG) response in smokers compared with nonsmokers. To investigate the effect of smoking on postprandial lipemia, a fat-rich mixed meal (837 kcal, 63 g of fat) was served to 12 healthy smokers and 12 controls with similar fasting lipoprotein profiles, body composition, and lifestyles. Blood was drawn before and 3, 4, 6, and 8 hours postprandially, and triglyceride-rich lipoprotein (TRL) fractions (chylomicrons, VLDL1, VLDL2, and IDL) were separated with density gradient ultracentrifugation. Pre- and postprandial TG, retinyl esters (RE), apolipoprotein B-48 (apoB-48) and B-100 (apoB-100) were measured in each fraction. Smokers showed a significantly increased postprandial TG response in chylomicrons, VLDL1, and VLDL2. The areas under the incremental curve (AUIC) of apoB-48 in chylomicrons (2.83 +/- 0.84 versus 0.56 +/- 0.17; P < .05) and VLDL1 (10.17 +/- 1.96 versus 2.95 +/- 2.44; P = < .01) were markedly higher in smokers than in controls. Changes of RE responses of all TRL fractions were consistent with those of apoB-48. Postprandial apoB-100 concentrations and lipolytic enzymes were similar between the two groups. In conclusion, smokers have the syndrome of impaired TG tolerance because of defective clearance of chylomicrons and their remnants. Prolonged residence time of atherogenic remnant particles may constitute a significant risk factor for CAD in smokers. PMID- 9351378 TI - Risk of coronary heart disease and activation of factor XII in middle-aged men. AB - Increased activity is known to be present in the extrinsic, intrinsic, and final common pathways of the hemostatic system in men at high risk of coronary heart disease (CHD), but the status of the contact system of coagulation in this condition is uncertain. Plasma levels of activated factor XII (XIIa), the initial product of contact activation, have therefore been measured by ELISA in 2464 men aged 51 to 62 years, clinically free of CHD, who were taking part in a prospective cardiovascular survey based in general medical practices. Statistically significant, independent, and positive associations of XIIa were found with serum cholesterol and triglyceride concentrations, blood pressure, body mass index, factor VII activity, plasma fibrinogen concentration, and tobacco smoking, all associated with CHD. Plasma XIIa also increased with recent alcohol intake. Men in the highest quintile of risk according to their conventional risk factors had a mean XIIa of 2.07 ng/mL (95% confidence interval 1.99-2.16), 31% higher than that of men in the lowest quintile (1.58; 95% confidence interval 1.51-1.65). Thus, the contact system of coagulation appears to be activated when CHD risk is increased. Furthermore, the independent associations of XIIa with the major conventional CHD risk factors and its broad range of values in the general population (0.1 to 12.5 ng/mL), combined with a relatively low day-to-day variability in individuals (the within-person component of its total variation being 14.7%), suggest its potential usefulness as a marker of atherosclerotic vascular damage. PMID- 9351379 TI - Thrombospondin-1 is a potent mitogen and chemoattractant for human vascular smooth muscle cells. AB - Thrombospondin-1 (TSP-1) is a matricellular protein that is present in negligible amounts in normal human vasculature but occurs in significant amounts in diseased vessels. In this study, we examined the effect of TSP-1 on DNA synthesis, proliferation, and migration in human vascular smooth muscle cells grown from saphenous vein. TSP-1 (0.1 to 30 micrograms/mL) elicited a concentration dependent increase in DNA synthesis under serum-free conditions. In combination with platelet-derived growth factor, TSP-1 induced a synergistic effect on DNA synthesis that was significantly higher than the additive effect of both agents. In proliferation assays, TSP-1 increased cell numbers by 50% relative to the serum-free controls over 14 days. In migration assays, conducted using modified Boyden chambers, TSP-1 (> or = 10 micrograms/mL) elicited marked chemotaxis to a degree equivalent to platelet-derived growth factor. The chemotactic response to TSP-1 (10 micrograms/mL) was abolished by the GRGDSP peptide but unaffected by the control GRGESP peptide, whereas neither peptide inhibited DNA synthesis stimulated by TSP-1. Inhibition of tyrosine kinase activity with genistein or tyrphostin A23 abolished DNA synthesis induced by TSP-1, and a neutralizing antibody to platelet-derived growth factor had no effect on DNA synthesis. Similarly, migration in response to TSP-1 was largely inhibited by these tyrosine kinase inhibitors. TSP-1 is a strong mitogen and chemoattractant for human vascular smooth muscle cells under serum-free conditions. The novel finding that TSP-1 is mitogenic for human cells contrasts with previous studies that have not shown any significant effect of TSP-1 itself on the growth of animal-derived smooth muscle cells. TSP-1 may play an important modulatory role in the local regulation of vascular smooth muscle function in vascular pathologies in humans. PMID- 9351380 TI - Sustained anti-CD4/CD8 treatment blocks inflammatory activation and intimal thickening in mouse heart allografts. AB - We evaluated inflammatory activation and vascular thickening in a heterotopic murine heart transplant model. C57BL/6J recipient mice received anti-CD4 therapy (days 1 to 4 after transplantation) or sustained, combined anti-CD4/CD8 therapy (days 1 to 4, weekly thereafter). Morphometric analysis of grafts (> 95 days) found the mean percentage of vessel occlusion to be 51.7% in allografts treated with anti-CD4, 8.3% in allografts treated with sustained anti-CD4/CD8, and 6.7% in isografts. Mean transcript levels of the adhesion molecules P-selectin, intercellular adhesion molecule 1 (ICAM-1), and leukocyte function-associated antigen 1 (LFA-1) and the cytokines interleukin 4 (IL-4), interferon-gamma (IFN gamma), inducible nitric oxide synthase (iNOS), allograft inflammatory factor 1 (AIF-1), and monocyte chemoattractant protein 1 (MCP-1) were measured with reverse transcription-polymerase chain reaction [RT-PCR] assays using deoxycytidine triphosphate radiolabeled with phosphorus 32 [32P-dCTP]. The assays were normalized against glyceraldehyde-3-phosphate dehydrogenase [G3PDH] Levels were found to be significantly higher in the anti-CD4 group than in the anti CD4/CD8 group. A strong correlation was also found between the percentage of luminal occlusion and the expression of these markers of inflammation (r = .92 .99, P < .0001). Sustained therapy involving proximal blockade of CD4 and CD8 interrupts pathways leading to inflammation and vascular thickening. However, long-term heart allografts in mice treated with a short course of anti-CD4 display an ongoing inflammatory cell activation that culminates in arteriosclerosis. This model may help examine the role of targeted immune factors using knockout mice to identify those causally involved in vessel thickening. PMID- 9351381 TI - Nitric oxide mediates LDL uptake in the artery wall in response to high concentrations of 17 beta-estradiol. AB - Female sex hormones are known to affect lipoprotein flux in the artery wall and atherosclerosis. However, the mechanisms of these artery wall effects are unclear. To examine the effect of 17 beta-estradiol (estradiol) on LDL uptake in the artery wall, we developed an isolated perfused rat carotid artery model from ovariectomized rats. LDL flux in the artery wall was measured by quantitative fluorescence microscopy before and after treatment with estradiol (0.001 to 10,000 nmol/L). Dose-response experiments showed no significant difference in the rate of LDL uptake when arteries were perfused with estradiol at physiological concentrations (0.001 to 1 nmol/L) compared with control perfusions. However, higher concentrations of estradiol (10 to 10,000 nmol/L) significantly increased the rate of LDL uptake in isolated arteries. Artery lumen volume significantly increased with perfusion of estradiol (1 to 100 nmol/L) but decreased after perfusions of higher concentrations of estradiol (1000 to 10,000 nmol/L). Additional studies were performed to examine mechanisms of estradiol-mediated increases in LDL uptake. The effect of estradiol (10 nmol/L) on the rate of LDL uptake was blocked by nitric oxide synthase inhibitors. However, the estrogen receptor antagonist tamoxifen did not block the effects of estradiol on the rate of LDL uptake. Our study indicates that modulation of LDL uptake in the artery wall by estradiol is concentration dependent. High concentrations of estradiol increase LDL uptake by production of endothelium-derived nitric oxide. These observations suggest that increased nitric oxide production compromises endothelial layer barrier function to increase LDL uptake in the artery wall. PMID- 9351382 TI - Large versus small unilamellar vesicles mediate reverse cholesterol transport in vivo into two distinct hepatic metabolic pools. Implications for the treatment of atherosclerosis. AB - Phospholipid liposomes are synthetic mediators of "reverse" cholesterol transport from peripheral tissue to liver in vivo and can shrink atherosclerotic lesions in animals. Hepatic disposal of this cholesterol, however, has not been examined. We compared hepatic effects of large (approximately equal to 120-nm) and small (approximately equal to 35-nm) unilamellar vesicles (LUVs and SUVs), both of which mediate reverse cholesterol transport in vivo but were previously shown to be targeted to different cell types within the liver. On days 1, 3, and 5, rabbits were intravenously injected with 300 mg phosphatidylcholine (LUVs or SUVs) per kilogram body weight or with the equivalent volume of saline. After each injection, LUV- and SUV-injected animals showed large increases in plasma concentrations of unesterified cholesterol, indicating mobilization of tissue stores. After hepatic uptake of this cholesterol, however, SUV-treated animals developed persistently elevated plasma LDL concentrations, which by day 6 had increased to more than four times the values in saline-treated controls. In contrast, LUV-treated animals showed normal LDL levels. By RNase protection assay, SUVs suppressed hepatic LDL receptor mRNA at day 6 (to 61 +/- 4% of control, mean +/- SEM), whereas LUVs caused a statistically insignificant stimulation. Hepatic HMG-CoA reductase message was also significantly suppressed with SUV, but not LUV treatment, and hepatic 7 alpha-hydroxylase message showed a similar trend. These data on hepatic mRNA levels indicate that SUVs, but not LUVs, substantially perturbed liver cholesterol homeostasis. We conclude that LUVs and SUVs mobilize peripheral tissue cholesterol and deliver it to the liver, but to distinct metabolic pools that exert different regulatory effects. The effects of one of these artificial particles, SUVs, suggest that reverse cholesterol transport may not always be benign. In contrast, LUVs may be a suitable therapeutic agent, because they mobilize peripheral cholesterol to the liver without suppressing hepatic LDL receptor mRNA and without provoking a subsequent rise in plasma LDL levels. PMID- 9351383 TI - Effect of coffee lipids (cafestol and kahweol) on regulation of cholesterol metabolism in HepG2 cells. AB - We studied the effect of the coffee diterpene alcohols, cafestol and kahweol, on cholesterol metabolism in HepG2 cells. Uptake of 125I-tyramine cellobiose-labeled LDL was decreased by 15% to 20% (P < .05) after 18 hours of preincubation with cafestol (20 micrograms/mL), whereas 25-hydroxycholesterol reduced uptake by 55% to 65% (P < .05). Degradation of LDL in the presence of cafestol was decreased by 20% to 30% (P < .05) under the same conditions. The effect of cafestol (20 micrograms/mL) on uptake and degradation of LDL was greatest (35% to 40%, P < .05) after 6 and 10 hours of preincubation, respectively. Furthermore, the effect of cafestol was also dependent on its concentration, and a significant decrease in the LDL uptake (19%) was observed at 10 micrograms/mL (P < .05). Specific binding of LDL was reduced by 17% (P < .05) and 60% (P < .05) after preincubation with cafestol (20 micrograms/mL) and 25-hydroxycholesterol (5 micrograms/mL) for 6 hours, respectively, compared with control cells. Analysis of LDL binding showed that cafestol reduced the number of binding sites for LDL on the cell surface (capacity) by 35% (P < .05). In contrast, no significant effect on the level of mRNA for the LDL receptor was observed after incubation with cafestol, whereas 25-hydroxycholesterol reduced the mRNA level for the LDL receptor by 40% to 50% (P < .05). A fusion gene construct consisting of a synthetic sterol regulatory element-1 (SRE-1) promoter for the human LDL receptor coupled to the reporter gene for chloramphenicol acetyltransferase (CAT) was transfected into HepG2 cells. No change was observed in CAT activity in SRE-1-transfected cells after incubation with cafestol, whereas 25-hydroxycholesterol reduced CAT activity by 30% to 40% (P < .05). Incorporation of [14C]acetate into unesterified cholesterol and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity were unaffected in cells incubated with cafestol as well as the cafestol kahweol mixture compared with control cells. Moreover, cafestol and the cafestol kahweol mixture did not promote increased incorporation of radiolabeled [14C]oleic acid into cholesteryl esters after short-term incubation compared with control cells. On the other hand, 25-hydroxycholesterol caused a 70% to 90% reduction of cholesterol synthesis (P < .05) and HMG-CoA reductase activity (P < .05), decreased HMG-CoA reductase mRNA level by 70% to 80% (P < .05), and promoted a twofold increase in cholesterol esterification (P < .05). Finally, no effect of the coffee diterpenes on bile acid formation was observed. These results suggest that cafestol (and kahweol) may reduce the activity of hepatic LDL receptors and thereby cause extracellular accumulation of LDL. PMID- 9351384 TI - Gender differences in intima-media permeability to low-density lipoprotein at atherosclerosis-prone aortic sites in rabbits. Lack of effect of 17 beta estradiol. AB - Premenopausal women are protected from coronary heart disease, and premenopausal nonhuman primates are protected from atherosclerosis, the underlying cause of coronary heart disease. Estrogen is thought to account for this protection in females, and part of this protection is independent of the effects on risk factors, including lipoprotein levels. This study considered the hypothesis that reduced intima-media permeability to low-density lipoproteins (LDL) may account for the protection from atherosclerosis and coronary heart disease in premenopausal females and that this effect might be mediated by estrogen. Intima media permeability to LDL was determined in male and female rabbits made hypercholesterolemic by feeding them 0.5% cholesterol for 8 days. The diet of half of the female rabbits was supplemented with 17 beta-estradiol (4 mg/d) during cholesterol feeding and the preceding 4 weeks. Estrogen treatment in the female rabbits did not influence the intima-media permeability to LDL. However, intima-media permeability to LDL for branch sites of the abdominal aorta and aortic arch (regions highly susceptible to atherosclerosis) was 43% and 38% lower, respectively, in male rabbits than in female rabbits: (2.93 +/- 0.39 microL/h/g, (n = 8), vs 6.28 +/- 0.86 microL/h/g, (n = 16), P < .001, and 4.69 +/ 0.28 microL/h/g, (n = 8) vs 7.57 +/- 0.75 microL/h/g, (n = 16), P < .02). In contrast, intima-media permeability to LDL in 7 of 8 aortic sites relatively resistant to atherosclerosis did not differ between male and female rabbits. These data suggest that the protection from atherosclerosis associated with female sex and estrogen is mediated by mechanism(s) other than reduction in intima-media permeability to LDL. PMID- 9351385 TI - HDL and ApoA prevent cell death of endothelial cells induced by oxidized LDL. AB - We have previously demonstrated that toxic doses of mildly oxidized LDL evokes in cultured cells a delayed and sustained rise of cytosolic [Ca2+], eliciting in turn irreversible cell damage and leading finally to cell death. HDL and delipidated apolipoprotein (apo). A prevented effectively the toxic effect of oxidized LDL to bovine aortic endothelial cells, in a time- and dose-dependent manner. The major part of the protective effect was mimicked by purified apoA-I, whereas purified apoA-II exhibited only very low protective activity. The protective effect was independent of the paraoxonase-linked HDL activity. The protective effect of HDL is independent of the contact of HDL with oxidized LDL, as shown by preincubation of oxidized LDL with HDL or apoA. In contrast, the protective effect was dependent on the integrity of apoA and on the contact of HDL with cells, thus suggesting that HDL acts directly on cells by enhancing their resistance against oxidized LDL. Preincubation experiments show that the protective effect is dependent on the duration of the contact of cells with HDL (maximal effect observed after 12 to 16 hours' preincubation), is also dependent on protein synthesis, and is persistent for at least 48 hours after the end of the contact of HDL with cells. Finally, effective concentrations of HDL inhibit the Ca2+ peak, which is directly involved in the cytotoxic effect of oxidized LDL, as shown by the inhibitory effect of Ca2+ chelators. All together, these results suggest that HDL, mainly apoA-I, increases the resistance of endothelial cells against oxidized LDL and prevents its toxic (apoptotic) effect by blocking the pathogenic intracellular signaling (culminating in sustained Ca2+ rise) involved in cell death. PMID- 9351386 TI - Lifetime smoking exposure affects the association of C-reactive protein with cardiovascular disease risk factors and subclinical disease in healthy elderly subjects. AB - Blood levels of C-reactive protein (CRP), a marker of inflammation, are related to cardiovascular disease risk. To determine cross-sectional correlates in the elderly, we measured CRP in 400 men and women older than 65 years and free of clinical cardiovascular disease at baseline as part of the Cardiovascular Health Study. Only 2% of the values were greater than 10 mg/L, the cut-point usually used to identify inflammation. CRP levels appeared tightly regulated, since there were strong bivariate correlations between CRP and the following: inflammation sensitive proteins such as fibrinogen (r = .52); measures of fibrinolysis such as plasmin-antiplasmin complex (r = .23); pack-years of smoking (r = .30); and body mass index (r = .24; all P values < or = .001). The association with pack-years was independent of the length of time since cessation of smoking. CRP levels were also associated with coagulation factors VIIc, IXc, and Xc; HDL cholesterol (negative) and triglyceride; diabetes status; diuretic use; ECG abnormalities; and level of exercise. Because of effect modification, two multiple linear regression prediction models were developed for CRP, one each for never smokers and ever smokers. An a priori physiologic model was used to guide these analyses, which disallowed the use of other inflammation-sensitive variables such as fibrinogen. In never smokers, the independent predictors were body mass index (+), diabetes status (+), plasmin-antiplasmin complex (+), and the presence of ECG abnormalities (+); this model predicted 15% of the CRP population variance. In ever smokers, the predictors were body mass index (+), plasmin-antiplasmin complex (+), pack-years of smoking (+), HDL cholesterol (-), and ankle-arm blood pressure index (-); this model predicted 42% of the population variance. We conclude that levels of CRP in the healthy elderly are tightly regulated and reflect lifetime exposure to smoking as well as level of obesity, ongoing level of fibrinolysis, diabetes status, and level of subclinical atherothrombotic disease. Moreover, exposure to smoking affects the relation of CRP to these other factors. PMID- 9351387 TI - Involvement of calcium and G proteins in the acute release of tissue-type plasminogen activator and von Willebrand factor from cultured human endothelial cells. AB - In this study, we investigated the role of Ca2+ and G proteins in thrombin induced acute release (regulated secretion) of tissue-type plasminogen activator (TPA) and von Willebrand factor (vWF), using a previously described system of primary human umbilical vein endothelial cells (HUVECs). The acute release of TPA and vWF, as induced by alpha-thrombin, was almost zero after chelation of Ca2+i, showing that an increase in [Ca2+]i was required. It did not matter whether the increase in [Ca2+]i came from an intracellular or extracellular Ca2+ source. Thrombin-induced release of TPA and vWF already started at low [Ca2+]i, around 100 nmol/L. Half-maximal release was found at a [Ca2+]i, of 261 nmol/L for TPA and at 222 nmol/L for vWF. The Ca2+ signal was transduced to calmodulin, as calmodulin inhibitors inhibited TPA and vWF release. The Ca2+ ionophore ionomycin dose dependently released vWF; half-maximal vWF release occurred at a [Ca2+]i of 311 nmol/L. In contrast, no TPA release was found at all below a [Ca2+]i of 500 nmol/L. Thus, below 500 nmol/L [Ca2+]i, an increase in [Ca2+]i alone was sufficient to induce vWF release but not sufficient to induce TPA release. Protein kinase C did not appear to be involved in TPA or vWF release, as neither an activator nor an inhibitor of protein kinase C significantly influenced release. Inhibition of phospholipase A2 also did not reduce thrombin-induced TPA and vWF release. The involvement of G proteins was studied by using both saponin permeabilized and intact cells. GDP-beta-S, which inhibits heterotrimeric and small G proteins, significantly inhibited thrombin-induced vWF and TPA release from permeabilized cells. AlF-4, which activates heterotrimeric G proteins, induced TPA and vWF release in both intact and permeabilized HUVECs. Preincubation of HUVECs with pertussis toxin significantly inhibited thrombin induced vWF release, due to inhibition of thrombin-induced Ca2+ influx. Pertussis toxin did not affect ionomycin-induced release. The inhibitory effect of pertussis toxin was less obvious in thrombin-induced TPA release, because it was counterbalanced by a positive effect of the toxin on TPA release. Thus, both inhibitory and stimulatory (pertussis toxin-sensitive) G proteins were involved in TPA release. Therefore, thrombin-induced acute release of TPA and vWF differed in two respects. First, below a [Ca2+]i of 500 nmol/L, an increase in Ca2+ was sufficient for vWF release but not for TPA release. Second, pertussis toxin sensitive G proteins were differentially involved in acute TPA and vWF release. PMID- 9351388 TI - The angiotensin-converting enzyme gene and the angiotensin II type I receptor gene as candidate genes for microalbuminuria. A study in nondiabetic and non insulin-dependent diabetic subjects. AB - Familial clustering of microalbuminuria with cardiovascular disease suggests a possible common genetic antecedent. We have tested the hypothesis that the angiotensin-converting enzyme (ACE) DD genotype and the angiotensin II type I receptor (AT1R) gene C allele represent the common link between microalbuminuria and coronary heart disease. The frequency of polymorphisms of the ACE and AT1R genes were investigated in 509 nondiabetic white subjects and in 86 non-insulin dependent diabetic white patients. There was no significant difference in albumin excretion rate between the genotypes in nondiabetic subjects on either a daytime or an overnight sample or in diabetic subjects expressed as a normalized albumin concentration on an untimed morning urine collection. We have found no evidence for an association between polymorphism of the ACE or AT1R genes and microalbuminuria in two groups of subjects without insulin-dependent diabetes. PMID- 9351389 TI - Gender-specific differences in the effects of testosterone and estrogen on the development of atherosclerosis in rabbits. AB - The aim of the present study was to investigate whether there are gender-specific differences in the effects of testosterone and estrogen on the process of atherogenesis. Thirty-two castrated male and 32 ovariectomized female rabbits were separated into 4 study groups of 8 males and 8 females each and received postoperatively a 0.5% cholesterol diet for 12 weeks. During this period either no hormones, estradiol (1 mg/kg body wt/week), testosterone (25 mg/kg body wt/week IMM), or estrogen combined with testosterone in above dosages were administered. Computerized morphometric analysis of the intimal thickening in the proximal aortic arch showed a significant inhibitory effect of estrogen in female and of testosterone in male animals (P < .05). In the group with combined treatment, the plaque size in both sexes was smaller than in the animals of the control group (P < .05). These differences were independent of changes in plasma lipid parameters. The incorporation of 5'-bromo-2'-deoxyuridine, associated with cell proliferation, into cells of the neointima was not significantly affected by the different hormone application regimens in males. In females, the incorporation rate was significantly lowered in the estrogen treated group compared with the control group (P < .05). Due to the observed differences in the sex specific atheroprotective effects of testosterone and estrogen, these data suggest that complex hormone interactions, which are independent of changes in plasma lipids, may play an important role in the process of atherogenesis. PMID- 9351390 TI - Fas is expressed in human atherosclerotic intima and promotes apoptosis of cytokine-primed human vascular smooth muscle cells. AB - The membrane protein Fas/Apo-1/CD95 signals programmed cell death or apoptosis in activated T lymphocytes. Vascular smooth muscle cells (SMCs) bear markers of programmed cell death or apoptosis in advanced atherosclerotic plaques that contain immune cells e.g., macrophages and T lymphocytes. This study tested the hypothesis that the Fas death-signaling pathway contributes to apoptosis of SMCs exposed to proinflammatory cytokines produced by these immune cells during atherogenesis. All atherosclerotic plaques examined (n = 14) contained immunoreactive Fas. The majority of the Fas+ SMCs localized in the intima of the plaques, whereas the medial SMCs expressed Fas antigen less prominently. Double staining for DNA fragments (TUNEL) and Fas or cell identification markers colocalized Fas with TUNEL+ SMCs in the areas that contained CD3+ T cells and CD68+ macrophages, suggesting a role for Fas in the induction of SMC apoptosis by activated T cells during atherogenesis. In culture, stimulation with interferon gamma, tumor necrosis factor-alpha, and interleukin-1 beta increased expression of Fas in SMCs. Incubation with an activating anti-Fas antibody triggered apoptosis of the cytokine-primed but not the untreated SMCs, as demonstrated by TUNEL and electrophoresis of oligonucleosomal DNA fragments. These data suggest that activation of the Fas death-signaling pathway contributes to the induction of SMC apoptosis during atherogenesis and furnish a mechanism whereby immune cells and their cytokines promote this cell death process related to vascular remodeling and plaque rupture. PMID- 9351392 TI - Topographical association between the cyclin-dependent kinases inhibitor P21, p53 accumulation, and cellular proliferation in human atherosclerotic tissue. AB - The cell cycle is controlled by cyclin-dependent protein kinases (CDKs). The activity of these enzymes is directed by inhibitors of CDKs. The 21-kD protein product (P21) of the WAF1/CIP1 gene, which can be transactivated by the protein product of the tumor suppressor gene p53, acts as an inhibitor of cyclin dependent kinases. To assess whether both P21 and p53 may play a role in the control of cellular proliferation in atherosclerotic lesions, the topographical association between p53, P21, and the proliferation marker MIB1/Ki-67, was analyzed by immunohistochemistry in human carotid atheromatous plaques of 26 patients. p53 immunoreactivity (IR) was present in 26 of 26 cases in the nuclei of virtually all cell types (macrophages [MPs], smooth muscle cells [SMCs], endothelial cells [ECs]) in areas with chronic inflammation in 71.08 +/- 8.28% of the nuclei. p53 staining in the control tissue from human coronary arteries was present in 0.3 +/- 0.45% of the cells (P < .002): P21-IR was present in 24 of 26 specimens in 64.38 +/- 10.13% of the cells (controls: 3.8 +/- 1.85%, P < .002) and localized to nuclei of MPs (CD68 positive) and SMCs (alpha-actin positive), as well as ECs of microvessels present in 21 specimens (21 of 21) and luminal ECs present in 18 specimens (16 of 18). As shown by double labeling, P21-IR colocalized with p53-IR in most MPs (24 of 24), intimal SMCs (22 of 24), ECs of microvessels (19 of 21), and luminal ECs (10 of 16). Interestingly, few p53 positive cells did not show simultaneous P21-IR, and, conversely, not all P21 positive cells demonstrated p53-IR. MIB1/Ki-67-positive cells were identified in 21 of 26 tissue specimens in 3.53 +/- 1.79% of the nuclei (controls: 0%, P < .002) and localized principally to MPs bordering the atheromatous lipid core (21 of 26) and to a few scattered SMCs (16 of 26), ECs of microvessels (13 of 21), and luminal ECs (2 of 18). Most importantly, none of the cells coexpressing P21 and p53 were positive for MIB1/Ki-67-IR, indicating the absence of proliferating activity. In summary, this study demonstrates that P21-IR is present in the atherosclerotic plaque and colocalizes with p53 in most MPs, SMCs, and ECs. The lack of proliferation markers in cells coexpressing p53 and P21 suggests that transcriptional activation of the WAF1/CIP1 gene by p53 may be involved in the control of cellular proliferation in advanced human atherosclerotic plaques. PMID- 9351393 TI - DNA fragmentation and ultrastructural changes of degenerating cells in atherosclerotic lesions and smooth muscle cells exposed to oxidized LDL in vitro. AB - Degeneration of smooth muscle cells in the fibrous cap of atherosclerotic lesions is an important factor in plaque rupture. Recent studies have suggested that many plaque cells are in a process of apoptosis as determined by positive deoxyribonucleotide-transferase-mediated dUTP end labeling. In this study, we demonstrate the existence of a colocalization between deoxyribonucleotide transferase-mediated dUTP end labeling-positive smooth muscle cells and oxidized LDL immunoreactivity in human carotid plaques. Oxidized LDL was found to induce deoxyribonucleotide-transferase-mediated dUTP end labeling positivity in cultured human smooth muscle cells, but only in the presence of tumor necrosis factor alpha and interferon-gamma. Electron microscopic analysis of cultured smooth muscle cells exposed to oxidized LDL in the absence of cytokines demonstrated cytoplasmic swelling and disruption of the plasma membrane, suggesting cell death by oncosis. Cells exposed to both oxidized LDL and cytokines were characterized by chromatin and cytoplasmic condensation compatible with cell death by apoptosis. These findings further support the notion that oxidized lipids play a role in plaque cell death. PMID- 9351391 TI - Altered compliance and residual strain precede angiographically detectable early atherosclerosis in low-density lipoprotein receptor deficiency. AB - BACKGROUND: This study was performed to detect changes in vascular biomechanical properties early in atherogenesis. METHODS AND RESULTS: Age- and weight-matched LDL-receptor deficient Watanabe hypercholesterolemic male rabbits (Group I: n = 11) and normal rabbits (Group II: n = 11) were studied. Fasting plasma lipoprotein concentrations, aortic angiography and intravascular ultrasound, in vivo aortic compliance evaluation, ex vivo aortic residual strain measurements, aortic lipid content and histopathology were determined. Plasma cholesterol was increased 9.8 fold and aortic cholesterol content was increased from 20 to 43 fold in Group I compared to Group II, respectively (P < .00005). Angiography revealed no stenoses in either group, whereas intravascular ultrasound and histological studies of Group I showed small circumferential plaques with < 10% cross-sectional area involvement. The residual strain in Group I was significantly increased in the ascending thoracic aorta (22.1 +/- 6.9% versus 10.4 +/- 3.2% in Group II, P < .0001), descending thoracic aorta (15.7 +/- 7.2% versus 4.8 +/- 1.3% in Group II, P < .0001), and abdominal aorta (18.0 +/- 4.8% versus 8.3 +/- 6.3% in Group II, P < .005). Changes in residual strain were inversely correlated with the aortic cholesterol content in the ascending thoracic aorta (r = -.72; P = -.001), descending thoracic aorta (r = -.95; P < .001), and abdominal aorta (r = -.51; P = .019). CONCLUSIONS: Early atherosclerosis in LDL-receptor deficient rabbits, undetectable by angiography yet observed by intravascular ultrasound imaging and histology, is associated with marked changes in ex vivo residual strain. Alterations in vascular biomechanical properties, associated with changes in cholesterol content, may have physiologic consequences and may be useful in detecting and quantitating early atherosclerosis. PMID- 9351394 TI - Tissue factor expression on macrophages in coronary plaques in patients with unstable angina. AB - Tissue factor is a membrane-bound glycoprotein that functions in the extrinsic pathway of blood coagulation by acting as a cofactor for factor VII, and the resulting complex leads to thrombin production in vivo. The purpose of the present study is to determine whether macrophages express tissue factor in human coronary atherosclerotic plaques. We examined directional coronary atherectomy specimens from 24 patients with unstable angina and 23 with stable exertional angina. In these specimens, macrophages were detected in 22 (92%) of 24 patients with unstable angina versus 12 (52%) of 23 with stable exertional angina (P = .003). The percentage of macrophage infiltration area was significantly larger in patients with unstable angina than in those with stable exertional angina (17 +/- 3% versus 6 +/- 2%, P = .008). The immunohistochemical double staining revealed the expression of tissue factor on macrophages in 18 (75%) of 24 patients with unstable angina versus 3 (13%) of 23 with stable exertional angina (P < .0001). Thrombus was identified in 20 (83%) of 24 patients with unstable angina versus 12 (52%) of 23 with stable exertional angina (P = .02). Fibrin deposition was mainly observed around macrophages expressing tissue factor in the patients with unstable angina. We have shown that tissue factor expression on macrophages was more frequent in coronary atherosclerotic plaques in patients with unstable angina. Tissue factor expressed on macrophages may play an important role in the thrombogenicity in coronary atherosclerotic plaques of these patients. PMID- 9351395 TI - Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow. AB - The ability of gene targeting in the mouse species presents a powerful tool to determine the role of specific molecules in vascular biology. Using a denuding injury procedure, we recently reported that intimal lesions can be induced in the carotid artery of outbred mice. The technical challenge associated with achieving complete denudation and the relatively small size of the developing lesions prompted us to design the present model of neointima formation and vascular remodeling in the carotid artery of the inbred FVB mouse strain. Complete ligation of the vessel near the carotid bifurcation induced rapid proliferation of medial smooth muscle cells, leading to extensive neointima formation in the presence of an endothelial lining. Thrombus formation was not observed except in the most distal part of the vessel adjacent to the ligature. At 4 weeks after ligation, luminal area was reduced by approximately 80% through a combination of decreased vessel diameter and neointima formation. Ultrastructural analysis provided evidence for cell death in the developing neointima as well as the remodeling media. The present model might be useful in identifying those genes important for neointima formation and vascular remodeling. PMID- 9351396 TI - Increased blood flow induces regression of intimal hyperplasia. AB - We have previously shown that high shear stress inhibits growth of developing neointima in a primate model of polytetrafluoroethylene (PTFE) graft healing. We used this model to test the hypothesis that increased shear stress can cause atrophy of an established neointima. High porosity PTFE grafts were inserted into the aorto-iliac circulation bilaterally in baboons. These grafts develop neointimal hyperplasia comprising smooth muscle cells and a luminal surface of confluent endothelium. Neointima was allowed to develop for 2 months. At that time 8 animals were sacrificed. In eight other animals blood flow in one of two grafts was increased by construction of a femoral arterio-venous fistula. These animals were sacrificed 2 months later (4 months after graft placement). At four months, intimal cross sectional area was smaller on the high shear stress side compared to the contralateral, normal shear stress side (2.53 +/- 0.75 versus 6.83 +/- 0.65 mm2, P < .05). Neointima from grafts exposed to 2 months normal shear stress followed by 2 months of high shear stress had regressed when compared to normal-shear stress grafts studied at 2 months (2.53 +/- 0.75 versus 4.56 +/- 0.68 mm2, P < .05). Morphometric analysis using transmission electron microscopy revealed that the decrease in intimal cross sectional area was attributable to a loss of both smooth muscle cells and matrix. Endothelial nitric oxide synthase was induced in high-flow graft intima. These observations support the conclusion that elevated shear stress can cause vessel wall atrophy. This process might be mediated by nitric oxide. PMID- 9351397 TI - Effect of streptozotocin-induced hyperglycemia on lipid profiles, formation of advanced glycation endproducts in lesions, and extent of atherosclerosis in LDL receptor-deficient mice. AB - Investigations into the mechanisms by which diabetes accelerates atherosclerosis have been hampered by the lack of suitable animal models. We hypothesized that streptozotocin-treated LDL receptor-deficient mice would be a good model of diabetic atherosclerosis because streptozotocin causes diabetes in the parent C57BL/6 strain and because in these mice diet-induced hypercholesterolemia leads to the formation of advanced atherosclerotic lesions throughout the aorta. Diabetes was induced in 18 mice by intraperitoneal injection of streptozotocin. Low-dose insulin was given subcutaneously to prevent excessive mortality and extreme elevations in triglyceride levels. The control group was subjected to sham injections. Both groups were fed a diet containing .075% cholesterol for six months. Average blood glucose was higher in the diabetic group than in the control group (257 +/- 67 mg/dL versus 111 +/- 7 mg/dL, P < 0.05). Although plasma cholesterol was similar (966 +/- 399 versus 1002 +/- 180 mg/dL) in both groups, VLDL cholesterol was higher whereas LDL cholesterol was lower in the diabetic group. Immunocytochemical analysis demonstrated significantly more advanced glycation end-product (AGE) epitopes in the artery wall of the diabetic group, whereas staining for oxidation-specific epitopes was similar in both groups. Sera of diabetic mice also contained significantly more IgG autoantibodies that bound to several AGE epitopes than did sera from control mice. Despite the presence of hyperglycemia, diabetic dyslipidemia, and enhanced AGE formation in the diabetic mice, both groups had a similar extent of atherosclerosis (diabetic, 17.3 +/- 5.2; control, 16.5 +/- 6.6% of the aortic surface). These data suggest that, at least under conditions of marked hypercholesterolemia; hyperglycemia and enhanced AGE formation do not contribute significantly to atherogenesis in LDL-/- mice. PMID- 9351398 TI - Expression of phospholipase A2 isoforms in human normal and atherosclerotic arterial wall. AB - LDL particles must be modified in the arterial wall to be taken up by macrophages at an excessive rate, leading to foam cell formation. Phospholipase A2 (PLA2) has been shown to modify LDL particles in vitro by degrading its phospholipids, resulting in enhanced uptake by macrophages. Reaction products of PLA2 are lysophospholipids and nonesterified fatty acids (mainly arachidonic acid), which are precursors of potent inflammatory mediators and which have been found in atherosclerotic regions of the arterial wall. To elucidate the expression of PLA2 in normal and diseased arteries, frozen tissue sections of human nonatherosclerotic mesenteric artery and carotid plaques were examined by immunohistochemistry using specific antibodies against secretory PLA2 types I and II and cytosolic PLA2 (85 kd). Secretory PLA2 type I was not detected. High expression of secretory PLA2 type II was found throughout the media in both normal and atherosclerotic artery specimens, in which smooth muscle cells dominated. Cytosolic PLA2 was found exclusively in diseased artery, mainly in the intima in regions with an inflammatory infiltrate consisting of macrophages and smooth muscle cells. Furthermore, both normal and atherosclerotic artery possessed substantial PLA2 activity. It is suggested that secretory PLA2 type II could play an important role in early atherogenesis because it is present in the preatherosclerotic arterial wall, where it may lead to LDL modification, foam cell formation, and activation of immune mechanisms. PMID- 9351399 TI - A partial estrogen receptor agonist with strong antiatherogenic properties without noticeable effect on reproductive tissue in cholesterol-fed female and male rabbits. AB - Estrogen replacement therapy retards the development of cardiovascular disease and osteoporosis in postmenopausal women. However, long-term unopposed use increases the risk of cancer in endometrium and possibly in breast. The racemic compound ormeloxifene, widely used in India as an antifertility agent, is a partial estrogen receptor agonist with antiosteoporotic properties. The present study was undertaken to investigate the effect of the L-enantiomer (levormeloxifene) and the d-enantiomer (d-ormeloxifene) on the development of atherosclerosis. In a short-term experiment (6 weeks), 4 x 10 ovariectomized female rabbits were fed a 0.25% cholesterol-enriched diet and the effect on plasma cholesterol levels was studied. In a long-term experiment (13 weeks), 4 x 15 ovariectomized female and 4 x 15 shamoperated male rabbits were maintained at a similar plasma cholesterol level of 25 mmol/L and the effect on undamaged and balloon-injured arterial wall was studied. In both experiments, the rabbits were treated with levormeloxifene, d-ormeloxifene, 17 beta-estradiol, or placebo, respectively. In the short-term experiment, levormeloxifene, in contrast to d ormeloxifene, significantly reduced plasma cholesterol by 30% compared with the placebo group. In the long-term experiment, levormeloxifene, in contrast to d ormeloxifene, significantly reduced atherosclerosis by 50% in the undamaged arterial wall of both female and male rabbits. Because these rabbits were cholesterol-clamped, the antiatherogenic effect was not mediated via plasma cholesterol lowering. Like estrogen, levormeloxifene did not inhibit atherosclerosis in the endothelium-denuded site of aorta. The antiatherogenic effects of levormeloxifene were thus similar to those of estrogen, but produced in the absence of any noticeable estrogenic effect on uterine or testicular tissue. PMID- 9351400 TI - Dose-response comparison of RRR-alpha-tocopherol and all-racemic alpha-tocopherol on LDL oxidation. AB - Much data have accrued in support of the concept that oxidation of LDL is a key early step in atherogenesis. The most consistent data with respect to micronutrient antioxidants and atherosclerosis appear to relate to alpha tocopherol (AT), the predominant lipid-soluble antioxidant in LDL. There are scant data on the direct comparison of RRR-AT and all-racemic (rac)-AT on LDL oxidizability. Hence, the aim of the present study was to examine the relative effects of RRR-AT and all-rac-AT on plasma antioxidant levels and LDL oxidation in healthy persons in a dose-response study. The effect of RRR-AT and all-rac-AT at doses of 100, 200, 400, and 800 IU/d on plasma and LDL AT levels and LDL oxidation was tested in a randomized, placebo-controlled study of 79 healthy subjects. Copper-catalyzed oxidation of LDL was monitored by measuring the formation of conjugated dienes and lipid peroxides over an 8-hour time course at baseline and again after 8 weeks. Plasma AT, lipid-standardized AT, and LDL AT levels rose in a dose-dependent fashion in both the RRR-AT and all-rac-AT groups compared with baseline. There were no significant differences in plasma, lipid standardized, and LDL AT levels between RRR-AT and all-rac-AT supplementation at any dose comparison. The lag phases of oxidation were significantly prolonged with doses > or = 400 IU/d of RRR-AT and all-rac-AT, as measured by conjugated dienes assay and at 400 IU/d of RRR-AT and 800 IU/d of both forms of AT by lipid peroxide assay. Again, there were no significant differences in the lag phase of oxidation at each dose for RRR-AT when compared with all-rac-AT. Also, there were no significant differences in LDL oxidation after in vitro enrichment of LDL with RRR-AT and all-rac-AT. Thus, supplementation with either RRR-AT or all-rac-AT resulted in similar increases in plasma and LDL AT levels at equivalent IU doses, and the degree of protection against copper-catalyzed LDL oxidation was only evident at doses > or = 400 IU/d for both forms. PMID- 9351401 TI - Egr-1 is activated in endothelial cells exposed to fluid shear stress and interacts with a novel shear-stress-response element in the PDGF A-chain promoter. AB - Exposure of vascular endothelial cells to fluid mechanical forces can modulate the expression of many genes involved in vascular physiology and pathophysiology. Here, we report that platelet-derived growth factor (PDGF) A-chain gene expression is induced at the level of transcription in cultured bovine aortic endothelial cells exposed to a physiologic level of steady laminar shear stress (10 dyn/cm2). 5' Deletion analysis of the human PDGF-A promoter revealed that a GC-rich region near the TATA box was required for shear-inducible reporter gene expression. This element conferred shear inducibility onto a heterologous promoter-reporter construct that was otherwise unresponsive to shear stress. The induction of PDGF-A expression by shear was preceded by rapid and transient induction in the expression of the immediate-early gene, egr-1, which binds to GC rich sequences. Gel shift studies indicated that shear-induced Egr-1 bound to the proximal PDGF-A promoter in a specific and time-dependent manner, displacing Sp1 from their overlapping recognition elements. Overlapping consensus binding sites for Egr-1 and Sp1 also appear in the proximal promoters of several other endothelial genes, including transforming growth factor-beta 1 and tissue factor, whose expression is modulated by shear stress. These findings define the Egr-1 binding site in the proximal PDGF-A promoter as a shear-stress-responsive element and suggest that shear-stimulated Egr-1 gene expression may be a unifying theme in the induction of various other endothelial genes exposed to biomechanical forces. PMID- 9351402 TI - Adipose tissue lipoprotein lipase and hormone-sensitive lipase. Contrasting findings in familial combined hyperlipidemia and insulin resistance syndrome. AB - The metabolism of free fatty acids (FFA) is altered in two common atherosclerosis promoting disorders: familial combined hyperlipidemia (FCHL) and insulin resistance syndrome (IRS). It has been suggested that these two conditions may have a common etiology. The enzymes lipoprotein lipase (LPL) and hormone sensitive lipase (HSL) are rate-limiting steps for the turnover of fatty acids in adipose tissue, because they hydrolyze extracellular triglycerides in lipoproteins (LPL) and intracellular triglycerides in adipocytes (HSL). The present study was undertaken to simultaneously determine the activities of LPL and HSL in subcutaneous adipose tissue from male patients with FCHL and IRS. LPL and HSL activity was investigated in 10 nonobese FCHL patients and compared with 10 matched healthy nonobese subjects, and in 8 essentially normolipidemic IRS patients (who did not have overt diabetes mellitus) and compared with 9 nonobese matched control subjects. LPL activity was 43% lower in patients with IRS (P < .0005), as compared with control subjects, but HSL activity was not significantly different in the two groups, On the other hand, HSL activity was decreased by 45% in FCHL patients (P < .01), as compared with control subjects, but LPL activity was not significantly different in FCHL patients and the control group. In conclusion, triglyceride metabolism in adipose tissue is altered in both FCHL and IRS. However, the abnormalities observed involve impaired function of LPL in IRS and impaired function of HSL in FCHL, suggesting separate etiologies for the altered lipolysis in these conditions, at least in male subjects. PMID- 9351403 TI - Evidence for an alpha-granular pool of the cytoskeletal protein alpha-actinin in human platelets that redistributes with the adhesive glycoprotein thrombospondin 1 during the exocytotic process. AB - In a previous study, we have demonstrated that the platelet adhesive glycoprotein thrombospondin-1 (TSP-1) interacts specifically with the cytoskeletal protein alpha-actinin in a solid-phase binding assay. Stored in the alpha-granules of platelets, TSP-1 is secreted during cell activation and binds to the plasma membrane promoting the platelet macroaggregate formation. However, the molecular mechanism by which TSP-1 reaches and binds to the platelet surface is to date unelucidated. alpha-Actinin is an actin-binding and actinin-cross-linking protein that is present in most cells and may act as a link between the bundles of F actin and the plasma membrane. In this study, we have investigated a possible interaction of alpha-actinin with TSP-1 in platelets by examining their respective subcellular location during the platelet activation process. By indirect immunofluorescence. alpha-actinin was found to display a granular staining in resting platelets similar to that of TSP-1. Performing postembedding immunogold labeling for electron microscopy, we detected the presence of alpha actinin throughout the cytoplasm, but the strongest gold staining was found in organelles identified as alpha-granules on the basis of their ultrastructure and TSP-1 content. With the use of double immunogold labeling on platelets at different stages of activation by thrombin, both alpha-actinin and TSP-1 were seen redistributing from the alpha-granules to the platelet surface via the open canalicular system (OCS). At the same time, the cytoplasmic alpha-actinin concentrated toward the plasma membrane, but no colocalization with the F-actin bundles was evidenced. Finally, preembedding immunogold labeling and immunoprecipitation of 125I-surface-labeled, thrombin-activated platelets further demonstrated that alpha-actinin was expressed on the plasma membrane in the absence of any detectable expression of actin and that it could from molecular complexes with TSP-1 on activated platelets. These results suggest that alpha actinin found to be present on the platelet surface together with TSP-1 originates in the alpha-granules by fusion of the alpha-granules with the plasma membrane during platelet exocytosis. PMID- 9351404 TI - Preventive effect of ritodrine hydrochloride and/or urinary trypsin inhibitor against lipopolysaccharide-induced preterm delivery in mice. AB - BACKGROUND: The purpose of this study was to confirm the preventive effect of ritodrine hydrochloride (ritodrine) alone or ritodrine plus urinary trypsin inhibitor (UTI) in a mouse model of preterm delivery. METHODS: On day 17 of pregnancy, female C3H/HeN mice impregnated by male B6D2F1 mice were given two intraperitoneal injections of lipopolysaccharide (LPS) (50 micrograms/kg) at a 3 hour interval, which induced a 100% incidence of preterm delivery within 25 hours of the second dose. Ritodrine (1, 3, or 10 mg/kg, p.o.), UTI (25 X 10(4) units/kg, i.p.), ritodrine (3 mg/kg, p.o.) plus UTI (25 x 10(4) units/kg, i.p.), distilled water (10 ml/kg, p.o.), or distilled water (10 mg/kg, p.o.) plus saline solution (10 ml/kg, i.p.) were administered to the pregnant animals 10 times at 1 hour intervals from 8:00 AM to 5:00 PM on day 18 of pregnancy. In addition, the preventive effect of ritodrine, UTI, or ritodrine plus UTI was examined on LPS induced contraction of uterine muscle strips isolated from pregnant mice on day 17 of gestation. RESULTS: The incidence of preterm delivery decreased significantly in a dose-dependent fashion with ritodrine treatment, and there was a significant and synergistic decrease after combined treatment with ritodrine plus UTI. The in vitro uterine contraction induced by LPS was significantly suppressed by both ritodrine and UTI. CONCLUSIONS: Combination therapy with ritodrine plus UTI may be helpful for preventing preterm delivery in humans without the cardiovascular side effects that often accompany treatment with ritodrine alone. PMID- 9351405 TI - Maternal serum screening for Down's syndrome on population basis. AB - BACKGROUND: The favorable attitude among the public towards prenatal diagnostics in Finland allowed us to start a trial on population basis when screening for Down's syndrome by maternal serum markers and age was introduced. METHODS: Screening by maternal serum markers for Down's syndrome was offered to all 17,200 pregnant women in the Helsinki area during the study period of 2.5 years. Screening due to advanced maternal age, 37 years or more, was continued as previously, and 1133 pregnant mothers used this option. Alpha-fetoprotein, human chorionic gonadotrophin, and during the first year also unconjugated estriol were used as markers. RESULTS: The uptake of serum screening was 84%. The proportion of false positive results i.e. risk for Down's syndrome, 1:350 or more at term, was initially 5.7%. After ultrasound scan 4.1% of the mothers remained 'screen positive'. The amniocentesis or chorionic villus sampling uptake was 98.4%. Ten out of eighteen cases of Down's syndrome were detected by maternal serum screening, sensitivity 56%, 95% CI 31-79%. Other chromosomal abnormalities were found in three cases, and there were four cases of mosaicisms confined to the placenta. These were trisomies 16, 7 and 2, and tetraploidy. Elevated serum alpha fetoprotein was found initially in 0.7% of the cases. One case of congenital nephrosis of the Finnish type and ten other, mainly structural, abnormalities were detected by high AFP. CONCLUSIONS: The screening was well received by the mothers. The detection rate of 56% is in the same range as in previous studies. Ultrasound scan before the test would effectively lower the false positive rate caused by incorrect timing. PMID- 9351406 TI - Iron supplementation in pregnancy: is less enough? A randomized, placebo controlled trial of low dose iron supplementation with and without heme iron. AB - BACKGROUND: The purpose of the present study was to evaluate the efficacy of low dose iron supplementation with and without a heme component, prescribed for women in the second half of pregnancy. METHOD: A randomized, double-blind, placebo controlled trial. Thirty-one women received a daily dose of 27 mg elemental iron in a product containing both heme iron and non-heme iron (Hemofer), 30 women received the same dose as pure non-heme iron with vitamin C (Collets jern med vitamin C), and 29 women received placebo. A double dummy technique was used to mask tablets. The women were tested for red cell indices and iron status markers (s-ferritin, s-iron, Total Iron Binding Capacity and erythrocyte protoporphyrin) throughout pregnancy and 8 and 24 weeks postpartum. The results were analyzed according to the 'intention to treat' principle. RESULTS: The hematological effects were equal in the two treatment groups. 25% of the supplemented women fell below 110 g/l in Hb vs 52% in the placebo group (p < 0.05); none fell below 100 g/l in the supplemented groups, 14% in the placebo group. Iron status was significantly better for all measured parameters in the heme iron group compared to placebo at the end of pregnancy. Differences between the other groups were only shown for some parameters, probably due to the small sample size. In the heme iron group there were fewer women with empty iron stores postpartum than at the start of pregnancy (from 14% to 8%), in the non-heme iron group there was a significant increase (from 3% to 27%), and in the placebo group the percentage of women with empty iron stores was more than doubled (from 21% to 52%). CONCLUSIONS: A daily dose of 27 mg elemental iron, containing a heme component, given in the second half of pregnancy, prevents depletion of iron stores after birth in most women. An equivalent dose of pure inorganic iron seems less effective, but the sample size in this study was too small to demonstrate significant differences between the two treatment groups. PMID- 9351407 TI - Ambulatory blood pressure monitoring in pregnancy induced hypertension. AB - OBJECTIVE: To determine the role of ambulatory blood pressure monitoring in the diagnosis of pregnancy-induced hypertension in women detected as hypertensive in the clinic by the conventional method. DESIGN: An observational study of ambulatory blood pressure monitoring. SETTING: A teaching hospital in Singapore. METHODS: One hundred and twenty-eight women between 28-37 weeks of pregnancy diagnosed to have non-proteinuric pregnancy-induced hypertension in the clinic had 24 hours ambulatory blood pressure monitoring. The mean systolic and diastolic BP, systolic and diastolic 'white coat effect' and the diastolic load were the main parameters noted. RESULTS: One hundred and twenty subjects had valid recordings. Only 46 (38.3%) were found to be truly hypertensive on ABP monitoring, using a mean diastolic pressure cut off of 85 mmHg. The 'white-coat effect' was seen in both groups of women--the hypertensives as well as the normotensives, although the magnitude of the white coat effect had poor correlation with the clinic diastolic BP. A cut-off value for diastolic load of 20 per cent was found to detect all hypertensives correctly (sensitivity 100%) with a modest false positive rate of 17.5 per cent. CONCLUSIONS: 'White-coat hypertension' is common in pregnancy and ambulatory blood pressure monitoring would be helpful in identifying the true hypertensive without requiring unnecessary hospitalization. PMID- 9351408 TI - Tick-borne relapsing fever and pregnancy outcome in rural Tanzania. AB - OBJECTIVE: To assess the impact of tick-borne relapsing fever (TBRF) on the outcome of pregnancy. DESIGN: Case control study of 137 pregnant women (cases) and 120 non-pregnant women (controls) with TBRF between 1985 and 1995. SETTING: A rural hospital in Tabora Region, Tanzania. RESULTS: Risk of birth during the attack of TBRF was 58.0%, with an extremely high perinatal mortality of 436 per 1000 births. The total loss of pregnancies including abortions was 475. per 1000. Case-fatality rate in pregnant women was 1.5%, compared to 1.7% in the non pregnant women. A Jarisch-Herxheimer reaction was seen in 1.5% of the cases and in 1.7% of controls. Relapse rate was 3.6%, compared to 1.7% in non-pregnant women. Pregnant women with TBRF show higher densities of spirochetes than non pregnant women (p < 0.001). The risk of delivery during the attack was positively correlated to increasing density of the spirochetemia (p < 0.001) and to gestational age (p < 0.001). Perinatal death was related to low birthweight (p < 0.001) and low gestational age (p < 0.001) and not to degree of spirochetemia. CONCLUSIONS: The extremely high perinatal mortality rate during an attack asks for prevention and early effective management of TBRF. This is a challenge where access to health services in rural areas of developing countries is hampered by many factors. PMID- 9351409 TI - Analysis of obstetric complications reported to the National Patient Insurance Association in Finland from 1987 to 1995. AB - BACKGROUND: The launch of the National Patient Insurance Association in 1987 offered a good starting point to evaluate obstetric claims in Finland. In order to obtain full compensation after patient injury, proof of malpractice is no longer required. Thus, the register of the Association offers a solid data base to analyze these injuries. METHODS: A nationwide descriptive study of obstetric claims reported to the National Patient Insurance Association from 1.5.1987 to 31.12.1995. The recorded statistical datafiles of the Association were used in the analysis. RESULTS: A total of 801 obstetric claims were analyzed. This comprised 2% of all (n = 39189) claims during the same time period. Nearly all injuries leading to claims (683/801, 85%) occurred during delivery. In all, 156 (19.5%) claims resulted in compensation. The total sum of compensation paid was $ 1.3 million. Most often (30/80 37.5%) and highest compensation ($ 0.8 million) was paid due to delay in the diagnosis of fetal asphyxia. In all, 118 (0.3%) malpractice trials in court have arisen from all claims to the Association during the study period; only two (1.7%) resulted from obstetric causes. CONCLUSIONS: The data indicate that most common and serious obstetric complications are associated with delay in the diagnosis of fetal asphyxia. Thus, from both the legal and the medical points of view, pertinent fetal monitoring during delivery and umbilical artery blood gas analysis after delivery in all risk deliveries should be obligatory in all delivery units. Finally, and perhaps most importantly, the patient insurance has effectively prevented obstetric malpractice trials in court. PMID- 9351410 TI - Preterm birth and cerebral palsy. Predictive value of pregnancy complications, mode of delivery, and Apgar scores. AB - BACKGROUND: Preterm infants are at 8 times higher risk than term infants for pre- and perinatal brain damage, resulting in cerebral palsy. In this paper we have analysed the influence of prenatal and birth-related risk factors on cerebral palsy in preterm infants. METHODS: In a register-based study, 175 preterm singleton infants with cerebral palsy, born in 1982-86, were compared with 687 controls matched by gestational age and year of birth. RESULTS: Statistically significant higher rates in cases were found in parity > or = 3 (22% vs. 16%, p < 0.05), Cesarean section (67% vs. 56%, p < 0.01), and low Apgar scores at 1 minute (45% vs. 36%, p < 0.05). By multivariate analyses, two variables remained statistically significant: parity > or = 3 (adjusted OR = 1.53 (95% CI 1.00 2.34), p < 0.05) and Cesarean section (adjusted OR = 1.57 (95% CI 1.07-2.32), p < 0.05). CONCLUSIONS: Pregnancy complications preceding preterm birth did not imply a higher risk of cerebral palsy. Delivery by Cesarean section was a prognostic factor for developing cerebral palsy, and the predictive value of Apgar scores was highly limited. PMID- 9351411 TI - Stillbirth and maternal well-being. AB - BACKGROUND: Stillbirth imposes severe strains on the mother. Little is known about the long-term well-being after such an experience. METHODS: The study population comprises 380 women who experienced stillbirth and 379 control women with a livebirth in 1991. Data were collected by a postal questionnaire in 1994 in a nationwide study in Sweden. The response rate was 84%. RESULTS: The index women stated more often than controls that they had an improved relationship with the baby's father at the time of follow-up than before the child's birth, the ratio of proportions (with 95% confidence interval) of women with an improved relationship being 1.8 (1.4-2.2). The corresponding figure for high satisfaction with home and family situation at the time of the survey was 1.3 (1.1-1.6) and 3.7 (1.6-8.3) for low satisfaction with the appreciation they encountered outside the home. The cumulative incidence of separation/divorce after a stillbirth was the same as after a livebirth, 8%. Marital status strongly modified the results; for single women the ratio of proportions (stillbirth compared to livebirth) for an improved relationship with the baby's father was 0.2 (0.0-1.4) and for high satisfaction with home and family situation 0.1 (0.0-0.9). CONCLUSIONS: Stillbirth increases satisfaction with the relationship with the baby's father and the home and family situation, but it decreases maternal satisfaction of appreciation by others outside of the home. This traumatic experience does not affect the risk of separation/divorce, but single women are at risk of social complications after stillbirth, and psychosocial support may be appropriate for this subgroup. PMID- 9351412 TI - Posttraumatic stress reactions after emergency cesarean section. AB - BACKGROUND: The study aimed at answering the following questions: Do women experience emergency cesarean section as traumatic? Do women experience any posttraumatic stress reactions or even posttraumatic stress disorder (PTSD) one to two months after emergency cesarean section? METHODS: Twenty-five consecutive women were interviewed a few days and one to two months after emergency cesarean section. RESULTS: Nineteen (76%) of the 25 women had experienced their delivery by emergency cesarean section as a traumatic event. One to two months postpartum none of these women met all the diagnostic criteria of PTSD. However, 13 women had various forms of posttraumatic stress reactions and in eight cases (33%) symptoms of serious posttraumatic intrusive stress reactions. CONCLUSIONS: The emergency cesarean section was in the majority of the cases experienced as a mental trauma. Although none of the women suffered from PTSD one to two months postpartum, one third had serious posttraumatic intrusive stress reactions. The concept of traumatic stress thus seems to be relevant for investigations of psychological aspects of emergency cesarean section. PMID- 9351413 TI - C-reactive protein in uncomplicated parturients delivered by cesarean section. AB - BACKGROUND: The purpose of this study was to determine normal values and evaluate clinical variables associated with postoperative C-reactive protein serum concentration after uncomplicated abdominal delivery. METHODS: C-reactive protein serum concentrations were determined serially by a quantitative immunoturbidimetric assay in 479 clinically non-infected parturients undergoing uncomplicated Cesarean section, before operation and during the first week of puerperium. Associations between clinical variables and maximal postoperative C reactive protein levels were examined by variance analysis. RESULTS: The preoperative values were significantly higher in parturients from whom samples were obtained after the onset of labor and/or rupture of the membranes (16 +/- 10 (s.d.) mg/l) compared with the preoperative values obtained from parturients operated with no labor and intact membranes (12 +/- 7 (s.d.) mg/l) (p < 0.0001). An overall significant increase was observed in C-reactive protein concentrations after delivery, peaking on the second postoperative day, compared with the preoperative values. In the parturients operated upon after the onset of labor or ruptured membranes, postoperative values were 24% higher up to the sixth postoperative day than in parturients operated upon with intact membranes and no labor. The onset of labor, duration of operation, the number of vaginal examinations and the duration of internal monitoring before operation were associated with the maximal postoperative C-reactive protein concentrations, but they explained only 9% of postoperative CRP level variation. CONCLUSIONS: The wide range of serum C-reactive protein concentrations during puerperium complicates its use as a simple marker in obstetric postoperative complications. However, after normal uncomplicated operative delivery, C-reactive protein concentrations rapidly decrease towards the baseline after the second to third postoperative day. PMID- 9351414 TI - Effects of prostaglandin treatment and paracervical blockade on postoperative pain in patients undergoing first trimester abortion in general anesthesia. AB - BACKGROUND: The postoperative analgesic efficacy of a paracervical blockade (PCB) as an adjunct to general anesthesia (GA) during outpatient abortion (dilatation and curettage) is unclear, and the present study was initiated to evaluate if PCB is of significant importance per- or postoperatively. METHODS: Two hundred women (aged 18-49 years) were assigned to one of four groups; group 1 received vaginal prostaglandin (PG) (PGE1, gemeprost 1 mg) for softening of the cervix preoperatively and surgery was performed in GA, group 2 received preoperative PG and surgery was performed in GA + PCB; group 3 did not receive PG treatment and surgery was performed in GA and group 4 were subjected to GA + PCB without preoperative PG. RESULTS: Women receiving preoperative prostaglandin treatment (groups 1 and 2) reported significantly higher pain intensity already preoperatively, but also postoperatively as compared to patients not treated with PG. The patients subjected to prostaglandin treatment (groups 1 and 2) also had a significantly higher consumption of analgesics as compared to non-PG treated groups (3 and 4). The addition of PCB did not influence pre- and postoperative pain intensity significantly or consumption of analgesics. Patients receiving PG also reported significantly more nausea than the others although nausea was of low intensity. Patients receiving PG were, however, discharged earlier than the others from the hospital. CONCLUSIONS: Preoperative treatment of the cervix with prostaglandins was associated with significantly higher pain intensity both pre- and postoperatively, and increased need for analgesics postoperatively and more intense nausea. PCB given just before surgery did not result in significant postoperative analgesia. More efficient techniques for pain control should be developed for women subjected to first trimester abortion with preoperative PG treatment. PMID- 9351415 TI - Oral contraceptive use among female elite athletes and age-matched controls and its relation to low back pain. AB - BACKGROUND: Exogenous and endogenous female sex steroids may influence the risk of low back pain. The fact that back pain is a very common symptom during pregnancy supports this theory. Back pain is also more common among female than male athletes. Oral contraceptives have been suggested to increase the risk of low back pain. OBJECTIVE: To evaluate whether the prevalence of low back pain is higher among oral contraceptive users than non-users and if it differs between women taking part in different sports. METHODS: A questionnaire was sent to female elite athletes in volleyball (n = 205), basketball (n = 150), and soccer (n = 361) as well as to age-matched controls (n = 113). The questionnaire comprised questions about age, constitution, occupation, parity and use of contraceptive method as well as previous and current back pain and possible consequences of the back problems. RESULTS: The response rate was 85%. Between 42% and 52% of the women in the different groups used oral contraceptives. The groups were similar in most background variables, except that the volleyball and basketball players were taller. The prevalence of current low back pain was between 21% and 34% in the different athlete groups with an average of 30%, whereas only 18% of the controls suffered from low back pain (p < 0.01). The prevalence of low back pain within each group, athletes as well as controls, was similar in women who used, and did not use oral contraceptives. CONCLUSIONS: This study does not support the theory that low back pain is affected by the use of oral contraceptives. Instead, constitutional factors and mechanical stress during intense physical activity is probably more important. PMID- 9351416 TI - Spirometric disorders in women with genital descensus. AB - BACKGROUND: We hypothesized that abnormalities in connective tissue, found in women with genital descensus, could impact their pulmonary function. METHOD: Therefore we compared lung flows and volumes between women with (n = 100) and without (n = 100) descensus. RESULTS: Patients exhibited highly significant decrements in all expiratory flows, especially in the peak expiratory flow (-35%) and other flows at large lung volumes. The forced vital capacity and forced expired volume at 1 second, but not their ratio, were also decreased (-16% and 17%, respectively). These differences were exaggerated in postmenopausal subjects and in patients with third degree of descensus, but did not depend on the presence of stress incontinence. Lung flows and volumes did not change between follicular and luteal phase of the cycle, either in patients or in controls. The forced vital capacity decreased with increasing years past the menopauses in patients (65 +/- 10 ml per year), but not in controls. CONCLUSION: In women with genital descensus deteriorations in lung ventilatory function were observed in association with the presence and duration of postmenopauses. PMID- 9351417 TI - Nephropathia Epidemica infection during first trimester of pregnancy--normal fetal outcome. PMID- 9351418 TI - Chorea gravidarum. PMID- 9351419 TI - Intramural ectopic pregnancy. Sonographic picture and its relation with adenomyosis. PMID- 9351420 TI - Intermembraneous polyhydramnios in a pregnancy with separated membranes. PMID- 9351421 TI - Factor V deficiency in pregnancy complicated by Rh immunization and placenta previa. A case report and review of the literature. PMID- 9351422 TI - Leiomyomatosis peritonealis disseminata in a postmenopausal woman. PMID- 9351423 TI - Prostate cancer screening--what's a physician to do? PMID- 9351424 TI - Homocysteine levels and cardiovascular disease. PMID- 9351425 TI - Prophylaxis after HIV exposure. PMID- 9351426 TI - Use of adhesive nasal strips for nasal obstruction. PMID- 9351427 TI - Repairing lacerations in children. PMID- 9351428 TI - Evaluation of recurrent thrombosis and hypercoagulability. AB - The primary mechanisms for coagulation were described more than a century ago. However, inherited disorders of coagulation have been recognized for only a few decades. Activated protein C resistance and other related protein defects account for many cases of hypercoagulability. Primary care physicians can initiate testing in individuals for whom there is a high degree of suspicion. Testing for specific disorders is best performed before initiation of anticoagulant therapy. Management often includes long-term warfarin therapy following initial anticoagulation with heparin. PMID- 9351429 TI - Homocysteine: a new risk factor for atherosclerosis. AB - The accumulating evidence for the role of homocysteine as a risk factor for atherosclerosis is persuasive. A high plasma homocysteine concentration induces pathologic changes in the arterial wall and thus is strongly associated with an increased risk of atherosclerosis, manifested as cardiovascular, cerebrovascular and peripheral vascular events. Studies are being conducted to determine whether lowering homocysteine levels prevents occlusive events. At present, testing for elevated homocysteine concentrations should be considered in patients with premature atherosclerosis or a strong family history of atherosclerosis, since hyperhomocysteinemia is a common risk factor in these patients. Treatment of hyperhomocysteinemia is straightforward and associated with minimal risk. This disorder is usually correctable with vitamin supplements containing folic acid. PMID- 9351430 TI - Local treatment of rectal cancer. AB - While abdominoperineal resection with permanent colostomy is still required for most distal [corrected] rectal cancers, sphincter-saving local treatment by means of local excision, electrocoagulation or endocavitary contact radiation can be used for some highly selected distal tumors. Local treatment avoids a permanent colostomy and is associated with much lower morbidity and mortality rates than abdominoperineal resection. Strict criteria for patient selection are essential to successful local treatment. Optimal candidates include patients exhibiting the following features of rectal cancer: a distal rectal cancer less than 8 cm from the anal verge; a tumor with a diameter of 3 cm or less; a tumor that is well to moderately well differentiated histologically, and a tumor that is limited to the bowel wall. Preoperative studies such as transrectal ultrasonography enhance the accuracy of preoperative staging. In properly selected patients, the results of local treatment are equivalent to those of abdominoperineal resection of comparable tumors. Close follow-up is essential, and tumor recurrence can be treated for cure by abdominoperineal resection. PMID- 9351431 TI - Epilepsy in pregnancy. AB - Family physicians who provide obstetric care may periodically encounter a patient with a history of epilepsy, which may manifest before or after pregnancy. In either case, several issues need to be addressed. Pregnant women with epilepsy may have an increased frequency of seizures, with the potential for resultant maternal and fetal morbidity and mortality. Teratogenic effects of antiepileptic drugs include craniofacial abnormalities and neural tube defects. Management strategies include the prenatal use of folic acid and vitamin K, monotherapy with a single antiepileptic drug, and obtaining at least monthly free serum drug levels. Fortunately, with close monitoring and proper management, more than 90 percent of pregnancies in women with epilepsy will be uncomplicated. PMID- 9351432 TI - Nail gun injuries of the hand. AB - Nail gun injury of the hand is commonly encountered among workers in the construction industry. Successful management requires a thorough understanding of this unique injury, the recognition of nail shaft barbs, and appropriate nail removal and wound care, with referral when indicated. If barbs are encountered, nail removal involves cutting off the head of the nail and extracting the nail in the direction of entry. PMID- 9351433 TI - ACP issues guidelines on the early detection of prostate cancer and screening for prostate cancer. PMID- 9351434 TI - Rapid assay for HIV drug resistance. PMID- 9351435 TI - Is there major involvement of the renin-angiotensin system in cardiac hypertrophy? PMID- 9351436 TI - Left ventricular stretch stimulates angiotensin II--mediated phosphatidylinositol hydrolysis and protein kinase C epsilon isoform translocation in adult guinea pig hearts. AB - Stretch of neonatal cardiomyocytes activates phospholipase C with production of inositol trisphosphate and diacylglycerol in part by formation of angiotensin II (Ang II). However, the response of this pathway to physical stimuli in the adult heart is poorly understood. Thus, in isovolumic perfused guinea pig hearts, we characterized stretch-mediated phosphatidylinositol (PI) hydrolysis and protein kinase C (PKC) isoform translocation using elevated diastolic pressure. Balloon dilatation (minimum diastolic pressure, 25 mm Hg) of the left ventricle (LV) stimulated PI hydrolysis. Pretreatment of stretched hearts with the specific angiotensin (AT1) receptor antagonist losartan abolished stretch-mediated accumulation of inositol phosphates. To examine PKC isoform expression and activation under these conditions, whole-heart extracts were examined by immunoblot analysis. Ang II translocated PKC epsilon to the particulate fraction. 4 beta-Phorbol 12-myristate 13-acetate but not an inactive congener translocated PKC epsilon to the particulate fraction and produced a decrease in myocardial contractile function. Mechanical stretch also translocated PKC epsilon to the particulate fraction; however, this was attenuated but not abolished by losartan. We conclude that in the adult heart, LV dilation produced stretch-mediated activation of phospholipase C, which resulted in PI hydrolysis and PKC epsilon activation in part by stimulation of the local renin angiotensin system. In contrast to stretch-mediated inositol phosphate accumulation, PKC epsilon translocation is not prevented by AT1 receptor blockade, indicating that this PKC isoform can be activated in response to mechanical deformation by an Ang II independent mechanism in the adult myocardium. PMID- 9351437 TI - Evidence that angiotensin II and lipoxygenase products activate c-Jun NH2 terminal kinase. AB - The effect of angiotensin II (Ang II) to activate c-Jun amino-terminal kinase (JNK) was studied in a Chinese hamster ovary fibroblast cell line overexpressing the rat vascular type-1a Ang II receptor (CHO-AT1a). Ang II treatment induced a time-dependent activation of JNK. Ang II (10(-7) mol/L) activated JNK activity, with a peak at 30 minutes (9.39 +/- 2.52-fold, n = 7, P < .02 versus control), which was maintained until 3 hours (2.7 +/- 0.65-fold, n = 3, P < .02 versus control). Ang II-induced JNK activation at 30 minutes was inhibited by a specific lipoxygenase (LO) pathway inhibitor, cinnamyl-3,4-dihydroxy-alpha-cyanocinnamate (1 mumol/L) by 87.5% (n = 4, P < .01 versus Ang II-induced JNK activity). The direct addition of 12-HETE also induced a time-dependent JNK activation. 12-HETE (10(-7) mol/L) activated JNK activity, with a peak at 10 minutes (3.43 +/- 0.87 fold, n = 6, P < .02 versus control), which remained elevated until 1 hour. These results suggest that the LO pathway is a mediator of Ang II-induced JNK activation. 15-HETE can also activate JNK at 5 minutes, but this activity was reduced at 30 minutes and could not be seen at 1 hour, indicating that the time course was different from that seen with 12-HETE. N-Acetylcysteine (NAC), an antioxidant, was used to perturb intracellular reactive oxygen intermediate (ROI) levels to assess the role of endogenous ROIs in regulating JNK activity. Pretreatment of cells with 500 mumol/L NAC for 1 hour attenuated approximately 50% of Aug II-induced JNK activation, suggesting that ROIs, at least partially, mediate Ang II-induced JNK activation. Furthermore, 12-HETE-induced JNK activation was reduced by approximately 90% by NAC. Finally, pertussis toxin completely blocked 12-HETE-induced JNK activation, suggesting that Gi-protein signaling participates in 12-HETE-induced effects. These results suggest that LO activation plays a role in mediating Ang II-induced JNK activation in part by altering the redox tone and Gi-protein signaling of cells. PMID- 9351438 TI - Leukemia inhibitory factor, a potent cardiac hypertrophic cytokine, activates the JAK/STAT pathway in rat cardiomyocytes. AB - Leukemia inhibitory factor (LIF) is a member of the interleukin-6 family of cytokines, which induces a wide range of responses in a variety of cells. The aim of this study was to investigate whether LIF induces cardiomyocyte hypertrophy and transmits signals through the JAK/STAT (indicating just another kinase/signal transducer and activator of transcription) pathway in primary cultured neonatal rat cardiomyocytes. LIF increased protein content and [3H]phenylalanine uptake in cardiomyocytes in a dose-dependent manner. LIF (10(3) U/mL) induced rapid tyrosine phosphorylation of gp130, JAK1, JAK2, STAT1, and STAT3 but not Tyk2 or STAT2. LIF also induced autokinase activity of JAK1 in a time-dependent manner. Gel shift assays for interferon gamma activation site/interferon-stimulated responsive element and sis-inducible element (SIE) revealed that LIF induced dimerization of STAT1 and STAT3 and formation of sis-inducing factor complexes, which subsequently interacted with SIE in the promoter. Preincubation with anti STAT1 and anti-STAT3 antibodies inhibited the binding of SIF complexes. In conclusion, LIF induces cardiac hypertrophy and directly stimulates the JAK/STAT pathway in cardiomyocytes. PMID- 9351439 TI - Increased expression of interleukin-1 beta and monocyte chemotactic and activating factor/monocyte chemoattractant protein-1 in the hypertrophied and failing heart with pressure overload. AB - Studies on the effects of proinflammatory cytokines on the heart suggest that they play some roles in the pathogenesis of congestive heart failure (CHF). To determine the involvement of proinflammatory cytokine in cardiac hypertrophy and CHF induced by mechanical overload, we investigated the expression of interleukin (IL)-1 beta and monocyte chemotactic and activating factor (MCAF)/monocyte chemoattractant protein-1 (MCP-1) in the left ventricle (LV) of Dahl salt sensitive (DS) rats that showed hypertrophy of the LV induced by hypertension and subsequently developed CHF. The IL-1 beta mRNA content in the LV of DS rats increased 3.9-fold when LV hypertrophy developed, and the increase reached 6.2 fold at the CHF stage compared with that of age-matched Dahl salt-resistant (DR) rats. The amount of IL-1 beta in the LV was positively correlated with the LV weight/body weight ratio. Most of the IL-1 beta immunoreactivity was localized in the endothelial cells and interstitial macrophages. The mRNA levels of MCAF in the LV increased 3.6-fold at 11 weeks and reached 4.8-fold at the CHF stage relative to the age-matched DR rats. MCAF protein was localized to the endothelial cells and interstitial macrophages. In DS rats, the number of interstitial macrophages increased diffusely throughout the LV. We suggest that increased chemokine expression, macrophage infiltration, and proinflammatory cytokine expression play some role in the pathogenesis of cardiac hypertrophy and failure induced by chronic mechanical overload. PMID- 9351440 TI - Activated RhoA stimulates c-fos gene expression in myocardial cells. AB - Rho regulates various cell functions, including cell morphology and motility. However, the functional role of Rho on the signaling pathway in myocardial cells (MCs) is unknown. In the present study, we attempted to explore the mode of Rho action for c-fos gene expression in MCs. Expression of the c-fos promoter/enhancer linked to the luciferase reporter gene (c-fos luciferase) was stimulated by the wild type of RhoA and the point-mutated active form of RhoA (RhoA Val14) but not the biologically inactive effector domain mutant of RhoA. Rho GDP dissociation inhibitor inhibited the action of RhoA on c-fos luciferase expression. The deletion analysis revealed that the c-fos serum response element (SRE) and the 12-O-tetradecanoylphorbol-13-acetate response element (TRE) mainly account for c-fos luciferase expression by RhoA Val14. The c-fos SRE mutant, which contains an intact binding site for the serum response factor but lacks the ternary complex factor binding site, was activated by RhoA Val14. The action of RhoA Val14 on c-fos luciferase expression was not inhibited by downregulation of protein kinase C, protein kinase C inhibitors, or tyrosine kinase inhibitors. These results indicate that activated RhoA stimulates c-fos gene expression through the c-fos SRE and TRE and that the signaling pathway from activated RhoA to the c-fos promoter/enhancer is independent of these inhibitor-sensitive pathways in MCs. PMID- 9351441 TI - Trophic effect of human pericardial fluid on adult cardiac myocytes. Differential role of fibroblast growth factor-2 and factors related to ventricular hypertrophy. AB - Pericardial fluid (PF) may contain myocardial growth factors that exert paracrine actions on cardiac myocytes. The aims of this study were (1) to investigate the effects of human PF and serum, collected from patients undergoing cardiac surgery, on the growth of cultured adult rat cardiac myocytes and (2) to relate the growth activity of both fluids to the adaptive changes in overloaded human hearts. Both PF and serum increased the rate of protein synthesis, measured by [14C]phenylalanine incorporation in adult rat cardiomyocytes (PF, +71.9 +/- 8.2% [n = 17]; serum, +14.9 +/- 6.5% [n = 13]; both P < .01 versus control medium). The effects of both PF and serum on cardiomyocyte growth correlated positively with the respective left ventricular (LV) mass. However, the magnitude of change with PF was 3-fold greater than with serum (P < .01). These trophic effects of PF were mimicked by exogenous basic fibroblast growth factor (FGF2) and inhibited by anti-FGF2 antibodies and transforming growth factor-beta (TGF-beta), suggesting a relationship to FGF2. In addition, FGF2 concentration in PF was 20 times greater than in serum. On the other hand, the LV mass-dependent trophic effect, present in both fluids, was independent of FGF2 concentration or other factors, such as angiotensin II, atrial natriuretic factor, and TGF-beta. These data suggest that FGF2 in human PF is a major determining factor in normal myocyte growth, whereas unidentified LV mass-dependent factor(s), present in both PF and serum, participates in the development of ventricular hypertrophy. PMID- 9351442 TI - Interaction of the heavy and light chains of cardiac myosin subfragment-1 with F actin. AB - The interaction of the heavy chain (HC) and the light chain (cdLC1) of cardiac S1 (cdS1) with F-actin was studied by cross-linking, Western blotting, and fluorescence polarization methods. Incorporation of cdLC1 in cross-linked products was examined by Western blots using the primary antibody against 71-74 residues of cdLC1. Cross-linking with zero-length, water-soluble reagent yielded three products with apparent molecular masses of 150, 160, and 210 kD. Like in the case of cross-linking of skeletal S1 with actin, these complexes included only HC of S1 and actin. The composition of the products were as follows: 150 kD, one HC of S1 cross-linked through a primary site (on the C-terminal of the 20-kD fragment) to the N-terminus of actin; 160 kD, one HC of S1 cross-linked through a secondary site (on the 50 kD fragment) to the N-terminus of actin; and 210 kD, one HC of S1 cross-linked through primary and secondary sites to two actins. Four additional products with apparent molecular masses of 66, 120, 185, and 235 kD contained cdLC1 and were identified as cdLC1 + actin, cdLC1 + HCS1, cdLC1 + actin + HCS1, and cdLC1 + two actins + HCS1, respectively. The same products were observed when cross-linking was performed in cardiac myofibrils incubated with cdS1. The production of cross-linked complexes of the heavy and light chain with actin decreased with an increase in the molar ratio of cdS1:actin. To test whether the orientation of myosin heads depended on a degree of occupation of thin filaments, myofibrils were irrigated with varying concentrations of cdS1. Fluorescence polarization measurements of cdS1 bound to individual I-bands revealed that the orientation depended on the concentration. PMID- 9351443 TI - Involvement of transcriptional and posttranscriptional mechanisms in cardiac overload-induced increase of B-type natriuretic peptide gene expression. AB - The induction of atrial and ventricular B-type natriuretic peptide (BNP) gene expression is one of the earliest events occurring during hemodynamic overload. To examine the molecular mechanisms for increased BNP gene expression during cardiac overload, we studied the induction of the BNP gene expression compared with that of atrial natriuretic peptide (ANP) in a modified perfused rat heart preparation. An increase in right atrial pressure of 5 mm Hg resulted in a 1.4 fold (P < .05) and 2.2-fold (P < .01) increase in BNP mRNA levels after 1 and 2 hours, respectively, whereas ANP mRNA levels remained unchanged. Stretching for up to 2 hours also significantly increased right atrial immunoreactive BNP (ir BNP) levels (from 15.8 +/- 2.2 to 20.1 +/- 1.2 ng/mg, P < .05). Actinomycin D (10 micrograms/mL), a transcriptional inhibitor, completely inhibited the stretch induced increase in atrial BNP mRNA levels at 1 hour (P < .05) and 2 hours (P < .001), whereas a protein synthesis inhibitor, cycloheximide (90 micrograms/mL), had no effect on basal or direct mechanical stretch-induced increase in right atrial BNP mRNA levels. Furthermore, we examined the role of tyrosine kinase and protein kinase C activities in acute mechanical stretch-induced increase in BNP synthesis. Tyrosine kinase inhibitors lavendustin A (1 mumol/L) and tyrphostin A25 (3 mumol/L) and protein kinase C inhibitors staurosporine (30 nmol/L) and chelerythrine (1 mumol/L) prevented the stretch-induced increase in right atrial ir-BNP concentrations at 2 hours. In addition, chelerythrine inhibited the increase of right atrial BNP mRNA levels stimulated by cardiac overload. These resuls demonstrate that the early increase of BNP mRNA levels by mechanical stretch results from increased transcriptional activation and is independent of protein synthesis. Our results also suggest that protein kinase C and tyrosine kinases activities may be involved in coupling cardiac overload to alterations in atrial BNP synthesis. PMID- 9351444 TI - Skeletal muscle sarcoplasmic reticulum Ca(2+)-ATPase gene expression in congestive heart failure. AB - Congestive heart failure leads to skeletal muscle abnormalities, one of which is a prolongation of sarcoplasmic reticulum Ca2+ flux. The purpose of this study was to determine whether skeletal muscle of spontaneous hypertensive and heart failure rats have alterations in the expression of the sarcoplasmic (or endoplasmic) reticulum Ca(2+)-ATPase (SERCA) gene. Northern analysis revealed that SERCA1, the predominant skeletal muscle isoform, was decreased by 45%, 43%, and 58% in the tibialis anterior, plantaris, and diaphragm muscles, respectively. Ribonuclease protection assay showed that the decrease was due to the adult isoform, SERCA1a, with minor changes in the alternatively spliced neonatal isoform, SERCA1b. There was no change in SERCA1 mRNA levels in gastrocnemius muscles. No change was found in SERCA2a (cardiac/slow skeletal isoform) mRNA or protein levels or in SERCA2b (smooth muscle isoform), dihydropyridine receptor, or alpha-actin mRNA levels in diaphragm muscle. Northern blot and ribonuclease protection assays showed that SERCA2a decreased 61% in the heart while the alternatively spliced isoform, SERCA2b, decreased 27%. Western analysis of the tibialis anterior, diaphragm, and gastrocnemius muscles showed a decrease in SERCA1 protein levels by 46%, 64%, and 42%, respectively, whereas sarcoplasmic reticulum Ca(2+)-ATPase activity, a functional correlate of SERCA expression, was decreased by 38%, 38%, and 40% in the same muscles, SERCA2 protein expression decreased by 36% in the failing heart. Decreases in both mRNA and protein suggest pretranslational control of SERCA1 expression, whereas the lack of decreased SERCA1 mRNA in gastrocnemius muscle suggests translational regulation. The decreased SERCA1 protein expression in all muscles studied probably contributes to contractile abnormalities related to excitation-contraction coupling function in heart failure. PMID- 9351445 TI - Unexpected and differential effects of Cl- channel blockers on the Kv4.3 and Kv4.2 K+ channels. Implications for the study of the I(to2) current. AB - The Kv4.3 K+ channel is thought to underlie the Ca(2+)-insensitive transient outward current (I(to1)) in ventricular myocytes of canine and human heart and to contribute to the I(to1) in rat myocytes. It has been suggested that there is a second component of the transient outward current in some species that is contributed by a Ca(2+)-activated Cl- current (known as I(to2)). The evidence for the existence of the I(to2) current is based, in part, on the pharmacological effects of various Cl- channel blockers. To test for possible interactions between these compounds and I(to1), the effect of several different Cl- channel blockers on the Kv4.3 channel was examined. The fenamates (niflumic and flufenamic acid) were found to have large effects on the position of the steady state inactivation curve of the Kv4.3 channel. The disulfonic stilbenes (DIDS and SITS) had markedly different effects and were found to greatly reduce the rate of recovery from inactivation of the Kv4.3 channel without large changes in the position of the activation and steady state inactivation curves. Both classes of drugs produced an apparent blockade of the Kv4.3 channel under some recording conditions. Surprisingly, the closely related Kv4.2 channel was found to be markedly less sensitive to these drugs. Caffeine was found to block both the Kv4.3 and Kv4.2 channels to a similar extent. These nonspecific drug effects have implications for the study of the two components of the transient outward current and suggest that purely pharmacological criteria cannot be used to define the physiological role of I(to2). PMID- 9351446 TI - Electrophysiological characterization of an alternatively processed ERG K+ channel in mouse and human hearts. AB - Mutants of HERG, the human form of ERG (the ether-a-go-go-related K+ channel gene), are responsible for some forms of the long-QT syndrome, an abnormality of cardiac repolarization. HERG was cloned from brain and has properties similar but not identical to the rapidly activating component of the native cardiac K+ channel current (Ikr). We identified in the mouse an alternatively processed form of ERG (MERG B) that is expressed abundantly in heart but only in trace amounts in brain. MERG B has a unique 36-amino acid NH2-terminal domain that is strongly basic and considerably shorter than the 376-amino acid NH2-terminal domain of HERG. When expressed in Xenopus oocytes, the kinetics of activation and deactivation of the MERG B current were best fit by a biexponential function, with the fast components dominant over the slow components. The fast component of activation had a mean tau value of 163 +/- 16 ms at -20 mV and 8 +/- 4 ms at +20 mV (n = 4). The fast component of deactivation had a mean tau value of 145 +/- 29 ms at -20 mV and 12 +/- 4 ms at -90 mV (n = 4). The MERG B current was blocked by the selective IKr blocker, dofetilide, with an IC50 of 54 nmol/L. In addition, we isolated HERG B, the human homologue of MERG B, which has electrophysiological characteristics qualitatively similar to those of MERG B. We have identified ERG B, an alternatively processed isoform of the ERG gene, expressed selectively in heart and with electrophysiological characteristics similar to those of native cardiac IKr. PMID- 9351447 TI - Ionic mechanisms of propagation in cardiac tissue. Roles of the sodium and L-type calcium currents during reduced excitability and decreased gap junction coupling. AB - In cardiac tissue, reduced membrane excitability and reduced gap junction coupling both slow conduction velocity of the action potential. However, the ionic mechanisms of slow conduction for the two conditions are very different. We explored, using a multicellular theoretical fiber, the ionic mechanisms and functional role of the fast sodium current, INa, and the L-type calcium current, ICa(L), during conduction slowing for the two fiber conditions. A safety factor for conduction (SF) was formulated and computed for each condition. Reduced excitability caused a lower SF as conduction velocity decreased. In contrast, reduced gap junction coupling caused a paradoxical increase in SF as conduction velocity decreased. The opposite effect of the two conditions on SF was reflected in the minimum attainable conduction velocity before failure: decreased excitability could reduce velocity to only one third of control (from 54 to 17 cm/s) before failure occurred, whereas decreased coupling could reduce velocity to as low as 0.26 cm/s before block. Under normal conditions and conditions of reduced excitability, ICa(L) had a minimal effect on SF and on conduction. However, ICa(L) played a major role in sustaining conduction when intercellular coupling was reduced. This phenomenon demonstrates that structural, nonmembrane factors can cause a switch of intrinsic membrane processes that support conduction. High intracellular calcium concentration, [Ca]i, lowered propagation safety and caused earlier block when intercellular coupling was reduced. [Ca]i affected conduction via calcium-dependent inactivation of ICa(L). The increase of safety factor during reduced coupling suggests a major involvement of uncoupling in stable slow conduction in infarcted myocardium, making microreentry possible. Reliance on ICa(L) for this type of conduction suggests ICa(L) as a possible target for antiarrhythmic drug therapy. PMID- 9351448 TI - Direct inhibition of expressed cardiac L-type Ca2+ channels by S-nitrosothiol nitric oxide donors. AB - NO donors have complex effects on Ca2+ currents in native cardiac cells, with reports of direct stimulation and indirect cGMP-mediated inhibition or stimulation. To investigate the molecular basis of these effects, we tested the effects of one class of NO donors, S-nitrosothiols (RSNOs), on expressed cardiovascular L-type Ca2+ channels (alpha 1C +/- beta 1a +/- alpha 2 or alpha 1C +/- beta 2a +/- alpha 2) in human embryonic kidney (HEK293) cells. The RSNO compounds we used were S-nitroso-N-acetylpenicillamine (SNAP, 5 to 10 nmol/L or 100 to 800 mumol/L), S-nitrosocysteine (SNC, 100 mumol/L or 1 mmol/L), and S nitrosoglutathione (GSNO, 1 mmol/L). Currents were measured using whole-cell patch recordings with 2 to 10 mmol/L Ba2+ as the charge carrier. SNAP reduced the amplitude of barium currents (IBa) through all the subunit combinations, with and EC50 of 360 mumol/L for alpha 1C + beta 1a channels. SNC or GSNO also inhibited IBa, albeit less potently. The inhibitory effect of SNAP was not affected by methylene blue (10 to 30 mumol/L) or 8-bromo-cGMP (200 to 400 mumol/L). The effects are relatively specific for Ca2+ channels, as expressed cardiac or skeletal muscle Na+ channels, which have a similar overall architecture, were barely affected by SNAP at concentrations as high as 1 mmol/L. We conclude that in the HEK293 expression system, the S-nitrosothiol NO donors inhibit L-type Ca2+ channels by a mechanism independent of cGMP. PMID- 9351449 TI - Attachment of meandering reentrant wave fronts to anatomic obstacles in the atrium. Role of the obstacle size. AB - Acetylcholine chloride (ACh) induces nonstationary meandering reentrant wave fronts in the atrium. We hypothesized that an anatomic obstacle of a suitable size prevents meandering by causing attachment of the reentrant wave front tip to the obstacle. Eight isolated canine right atrial tissues (area, 3.8 x 3.2 cm) were mounted in a tissue bath and superfused with Tyrode's solution containing 10 to 15 mumol/L ACh. Holes with 2- to 10-mm diameters were sequentially created in the center of the tissue with biopsy punches. Reentry was induced by a premature stimulus after eight regular stimuli at 400-ms cycle length. The endocardial activation maps and the motion of the induced reentry were visualized dynamically before and after each test lesion using 509 bipolar electrodes. In the absence of a lesion (n = 8), the induced single reentrant wave front, in the form of a spiral wave, meandered irregularly from one site to another before terminating at the tissue border. Holes with 2- to 4-mm diameters (n = 6) had no effect on meandering. However, when the hole diameters were increased to 6 mm (n = 8), 8 mm (n = 8), and 10 mm (n = 6), the tip of the spiral wave attached to the holes, and reentry became stationary. Transition from meandering to an attached state converted the irregular and polymorphic electrogram to a periodic and monomorphic activity with longer cycle lengths (101 +/- 11 versus 131 +/- 9 ms for no hole versus 10-mm hole, respectively; P < .001). Regression analysis showed a significant positive linear correlation between the cycle length of the reentry and the hole diameter (r = .89, P < .01) and between the cycle length of the reentry and the excitable gap (r = .89, P < .05). We conclude that a critically sized anatomic obstacle converts a nonstationary meandering reentrant wave front to a stationary one. This transition converts an irregular "fibrillation-like" activity into regular monomorphic activity. PMID- 9351450 TI - Bradykinin B2-receptor activation augments norepinephrine exocytosis from cardiac sympathetic nerve endings. Mediation by autocrine/paracrine mechanisms. AB - We determined whether local bradykinin production modulates cardiac adrenergic activity. Depolarization of guinea pig heart sympathetic nerve endings (synaptosomes) with 1 to 100 mmol/L K+ caused the release of endogenous norepinephrine (10% to 50% above basal level). This release was exocytotic, because it depended on extracellular Ca2+, was inhibited by the N-type Ca(2+) channel blocker omega-conotoxin and the protein kinase C inhibitor Ro31-8220, and was potentiated by the neuronal uptake-1 inhibitor desipramine. Typical of adrenergic terminals, norepinephrine exocytosis was enhanced by activation of prejunctional angiotensin AT1-receptors and attenuated by adrenergic alpha 2 receptors, adenosine A1-receptors, and histamine H3-receptors. Exogenous bradykinin enhanced norepinephrine exocytosis by 7% to 35% (EC50, 17 nmol/L), without inhibiting uptake 1. B2-receptor, but not B1-receptor, blockade antagonized this effect. The kininase II/angiotensin-converting enzyme inhibitor enalaprilat and the addition of kininogen or kallikrein enhanced norepinephrine exocytosis by approximately equal to 6% to 40% (EC50, 20 nmol/L) and approximately equal to 25% to 60%, respectively. This potentiation was prevented by serine protease inhibitors and was antagonized by B2-receptor blockade. Therefore, norepinephrine exocytosis is augmented when bradykinin synthesis is increased or when its breakdown is inhibited. This is the first report of a local kallikrein-kinin system in adrenergic nerve endings capable of generating enough bradykinin to activate B2-receptors in an autocrine/paracrine fashion and thus enhance norepinephrine exocytosis. This amplification process may operate in disease states, such as myocardial ischemia, associated with severalfold increases in local kinin concentrations. PMID- 9351451 TI - Pulsatile stretch stimulates superoxide production and activates nuclear factor kappa B in human coronary smooth muscle. AB - There is increasing evidence that oxidative stress is of pathophysiological importance in cardiovascular disease. Mechanical forces such as pulsatility may also contribute. Using human coronary artery smooth muscle cells (HCAS), we tested the hypothesis that stretch-induced cell proliferation is associated with oxidative stress. Stretch induced DNA synthesis in HCAS, and this was prevented by the antioxidants N-acetylcysteine and pyrrolidinedithiocarbamate (PDTC). Pulsatile stretch also increased superoxide production from HCAS in a time- and stretch dependent manner. Stretch-induced superoxide production was inhibited by diphenyleneiodoniumchloride, an NADPH oxidase inhibitor, and p chloromercuriphenylsulfonic acid, an NADH oxidase inhibitor, but not by the xanthine oxidase inhibitor oxypurinol or the cyclooxygenase inhibitor indomethacin. In electrophoretic mobility shift assays, tumor necrosis factor alpha activated nuclear factor-kappa B (NF-kappa B) with a peak at approximately 3 hours, whereas pulsatile stretch showed sustained activation during stimulation for up to 24 hours. The sustained activation of NF-kappa B was abolished by cotreatment with N-acetylcysteine or PDTC. Furthermore, treatment of HCAS with antisense p65 and p50 oligodeoxynucleotides of NF-kappa B inhibited stretch induced DNA synthesis. We propose that pulsatile stretch increases oxidative stress and, in turn, promotes DNA synthesis via NF-kappa B in cultured human coronary artery smooth muscle cells. PMID- 9351452 TI - Macrophage-dependent regulation of syndecan gene expression. AB - Heparan sulfates in the extracellular matrix are required for a variety of biological processes, including cellular response to heparin-binding growth factors. However, little is known regarding the regulation of their expression and composition under pathophysiological conditions. In the present study, we have investigated the regulation of expression of two key heparan sulfate chain carrying core proteins, syndecan-1 and syndecan-4, in a mouse/rat infarct model of tissue injury and repair. Induction of myocardial infarction was associated with a prompt increase in expression of both syndecan genes. Although infiltrating macrophages accounted for a substantial increase in syndecan expression, increased expression was noted in the levels of syndecan-1 mRNA in endothelial cells and syndecan-4 mRNA in cardiac myocytes. This increase in expression was limited to the immediate peri-infarct region and was absent from remote areas of the left or right ventricles. The influx of blood-derived macrophages in the heart correlated with the appearance of PR-39 peptide, which has previously been shown to increase syndecan expression in vitro. Studies in the op/op mice strain (which demonstrates sharply reduced levels of circulating monocytes) showed that myocardial infarction was associated with markedly reduced levels of macrophage influx and corresponding reduction in the expression of PR 39 and both syndecan genes. Pretreatment of op/op mice with granulocyte macrophage colony-stimulating factor restored myocardial macrophage content with corresponding restoration of PR-39/syndecan expression. In summary, myocardial infarction is associated with a distinct spatial and temporal pattern of syndecan 1 and -4 gene expression, which is induced by an influx of blood-derived macrophages. PMID- 9351453 TI - Expression, function, and regulation of E-type prostaglandin receptors (EP3) in the nonischemic and ischemic pig heart. AB - The action of prostacyclin, prostaglandin E1 (PGE1), and their mimetics on myocardial function includes changes in contractility, electrophysiological properties, and protection from injury caused by transient myocardial ischemia. This study was undertaken to investigate the basic properties of myocardial E type prostaglandin (EP) receptors. Ligand binding studies using an enriched preparation of sarcolemmal membranes prepared from pig hearts revealed a single class of binding sites for [3H]PGE1, with a Kd of 3.7 nmol/L and a Bmax of 92 fmol/mg protein. Competition experiments indicated highest affinity for EPs, suggesting an EP receptor. In addition, the EP receptor subtype-selective agonists sulprostone (EP1 and EP3) and M&B 28.767 (EP3) were active, suggesting the presence of an EP3 receptor subtype. PGE1 stimulated sarcolemmal GTPase and inhibited sarcolemmal adenylyl cyclase activity, indicating EP3 receptor coupling to an inhibitory G protein (Gi). Additional in vivo experiments showed that intracoronary infusion of PGE1 (1 nmol/min) decreased isoprenaline-stimulated left ventricular contractile activity without altering systemic vascular resistance. This inhibition of beta-adrenergic effects is compatible with the known myocardial anti-ischemic action of prostaglandins. Further experiments examined EP3 receptor density and G-protein coupling in sarcolemma from ischemic and reperfused ischemic myocardium. In anesthetized open-chest minipigs, occlusion of the left anterior descending coronary artery for 60 minutes increased EP3 receptor density by 50%, whereas receptor affinity was unchanged. This upregulation was prevented by pretreatment with colchicine (2 mg/kg i.v.), indicating microtubule-dependent receptor externalization. Northern hybridization showed comparable EP3 receptor mRNA expression in control and ischemic myocardium. The increase of receptor protein was reversed during 60 minutes of reperfusion. G-protein coupling proved to be intact in ischemic and reperfused ischemic myocardial tissue, as shown by preserved GTP-gamma-S-induced decrease of [3H]PGE1 binding. These data demonstrate for the first time that myocardial receptors for PGE1 belong to the EP3 subtype. The properties of this receptor include inhibition of adenylyl cyclase and upregulation during regional myocardial ischemia, suggesting an involvement in the anti-ischemic activity of E and I-type prostaglandins. PMID- 9351454 TI - Angiotensin II-induced leukocyte adhesion on human coronary endothelial cells is mediated by E-selectin. AB - Clinical data suggest a link between the activation of the renin-angiotensin system and cardiovascular ischemic events. Leukocyte accumulation in the vessel wall is a hallmark of early atherosclerosis and plaque progression. E-Selectin, vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) are adhesion molecules participating in mediating interactions between leukocytes and endothelial cells and have been found to be expressed in athero sclerotic plaques. We investigated whether angiotensin II, the effector of the renin-angiotensin system, influences the endothelial expression of E-selectin, VCAM-1, and ICAM-1. In coronary endothelial cells derived from explanted human hearts, angiotensin II (10(-11) to 10(-5) mol/L) induced a concentration dependent increase in E-selectin expression. The effect was measured by cell ELISA and duplex reverse-transcription polymerase chain reaction (RT-PCR) and reached its maximum at 10(-7) mol/L. Angiotensin II induced only a small increase in E-selectin expression in cardiac microvascular endothelial cells. VCAM-1 and ICAM-1 were not affected by angiotensin II stimulation. In addition, the effect of angiotensin II-induced E-selectin expression on leukocyte adhesion was quantified under flow conditions. Angiotensin II (10(-7) mol/L) increased leukocyte adhesion significantly to 67% of the maximal effect by tumor necrosis factor-alpha at a wall shear stress of 2 dyne/cm2. This adhesion was found to be E-selectin dependent, as demonstrated by blocking antibodies. The AT1-receptor antagonist DUP 753 significantly reduced E-selectin-dependent adhesion, whereas the AT2-receptor antagonist PD 123177 had no inhibitory effect. In addition, only AT1-receptor, but not AT2-receptor, mRNA could be detected by RT-PCR in coronary endothelial cells. Therefore, it is suggested that AT1 receptors mediate the effects of angiotensin II on E-selectin expression and leukocyte adhesion on coronary endothelial cells. PMID- 9351455 TI - Phenylephrine-induced Ca2+ oscillations in canine pulmonary artery smooth muscle cells. AB - Modulation of [Ca2+]i in response to receptor activation is a critical determinant of vascular smooth muscle tone. In this study, we examined the effect of continuous stimulation of alpha 1-adrenoceptors with phenylephrine (PE) on [Ca2+]i in single pulmonary artery smooth muscle cells (PASMCs) cultured from explants of canine intrapulmonary artery. Fura 2-loaded PASMCs pretreated with propranolol (5 mumol/L) were continuously superfused with PE at 37 degrees C on the stage of an inverted fluorescence microscope, and [Ca2+]i was measured using a dual-wavelength spectrofluorometer. Resting values of [Ca2+]i were 96 +/- 4 nmol/L. PE (10 mumol/L) stimulated oscillations in [Ca2+]i at a frequency of 1.35 +/- 0.07/min, which reached a peak [Ca2+]i of 650 +/- 26 nmol/L (n = 69 cells). The oscillations lasted for > 30 minutes and were constant in amplitude and frequency. Both the amplitude and frequency of PE-induced [Ca2+]i oscillations increased in a dose-dependent (3 x 10(-8) to 10(-4) mol/L) manner. Pretreatment with the alpha 1-adrenoceptor antagonist prazosin (50 nmol/L) or removal of extracellular Ca2+ abolished the repetitive [Ca2+]i oscillations induced by PE. The voltage-operated Ca2+ channel blockers nifedipine (1 mumol/L) and verapamil (1 mumol/L) had no effect on the [Ca2+]i oscillations. In contrast, inhibition of phospholipase C with U73122 (10(-7) to 10(-5) mol/L) attenuated the oscillations in a dose-dependent fashion. The nonselective protein kinase inhibitor staurosporine (10(-9) to 10(-7) mol/L) had a minimal inhibitory effect on the oscillations. Caffeine (30 mmol/L) and thapsigargin (1 mumol/L) abolished the oscillations, whereas pretreatment with ryanodine (1 to 100 mumol/L) had no effect. In freshly dispersed PASMCs, PE (10 mumol/L) induced oscillations in [Ca2+]i similar to those observed in cultured cells, and patch-clamp experiments revealed oscillations in membrane potential. These results indicate that PE induces [Ca2+]i oscillations in PASMCs via stimulation of alpha 1-adrenoceptors coupled to phospholipase C activation. Voltage-operated Ca2+ channels and protein kinases are not required for the oscillations. The requirement for extracellular Ca2+ and intracellular Ca2+ stores indicates that both Ca2+ influx and intracellular Ca2+ release play a role in the maintenance of the oscillations. PMID- 9351456 TI - Coagulation factor Xa induces endothelium-dependent relaxations in rat aorta. AB - The relaxing effect of coagulation factor Xa on phenylephrine-contracted rat aortic rings was compared with the effect of thrombin and trypsin. All three proteases induced a dose-dependent relaxation in the presence of an intact endothelium. EC50 values were 3 +/- 1, 24 +/- 9, and 16 +/- 1 nmol/L for thrombin, trypsin, and factor Xa, respectively. Whereas thrombin induced rapid relaxations followed by partial recontraction, trypsin and factor Xa induced slower sustained effects. Factor Xa-induced relaxations were not affected by hirudin at high concentrations (1 mumol/L) but were abolished by DX9065A, a specific inhibitor of the catalytic activity of factor Xa. Furthermore, no relaxations to factor Xa could be elicited in the presence of the NO synthase inhibitor N omega-nitro-L-arginine methyl ester (100 mumol/L), whereas relaxations were not altered in the presence of the inactive enantiomer N omega nitro-D-arginine methyl ester (100 mumol/L). Addition of factor Xa together with thrombin induced relaxations that were larger than those induced by thrombin alone, whereas factor Xa had no additional effects on trypsin-induced relaxations. Further-more, factor Xa relaxed thrombin-desensitized aortic rings but was ineffective in trypsin-desensitized tissues. These data suggest that factor Xa acts on a cleavable endothelial receptor that induces NO release, resulting in the relaxation of precontracted rat aortic rings. Factor Xa does not act through endothelial thrombin receptors but may activate another cleavable trypsin-sensitive receptor. PMID- 9351457 TI - Urokinase but not tissue plasminogen activator mediates arterial neointima formation in mice. AB - To define the role of the plasminogen activators (PAs) tissue PA (t-PA) and urokinase PA (u-PA) in vascular wound healing, neointima formation and reendothelialization were evaluated after electric or mechanical arterial injury in mice with a single or combined deficiency of t-PA (t-PA-/-) and/or u-PA (u-PA /-). In both models, neointima formation and neointimal cell accumulation were reduced in u-PA-/- and in t-PA-/-/u-PA-/- arteries but not in t-PA-/- arteries. The electric injury model was used to characterize the underlying cellular mechanisms. Topographic analysis of vascular wound healing in electrically injured wild-type and t-PA-/- arteries revealed a similar degree of migration of smooth muscle cells from the noninjured borders into the necrotic center. In contrast, in u-PA-/- and t-PA-/-/u-PA-/- arteries, smooth muscle cells accumulated at the uninjured borders but failed to migrate into the necrotic center. Cultured u-PA-/- but not t-PA-/- smooth muscle cells also failed to migrate in vitro after scrape wounding. Proliferation of smooth muscle cells was not affected by PA deficiency. Reendothelialization after electric injury was similar in all genotypes. In situ analysis revealed markedly elevated u-PA zymographic activity, mRNA, and immunoreactivity in smooth muscle cells, endothelial cells, and leukocytes within 1 week after injury, eg, when cells migrated into the wound. Thus, u-PA plays a significant role in vascular wound healing and arterial neointima formation after injury, most likely by affecting cellular migration. PMID- 9351458 TI - Dual role for nitric oxide in the regulation of plasma volume and albumin escape during endotoxin shock in conscious rats. AB - To assess the role of nitric oxide (NO) produced by the constitutive (cNOS) and inducible NO synthase (iNOS) in the regulation of vascular functions, we compared the effects of aminoguanidine, a relatively selective inhibitor of iNOS, and NG nitro-L-arginine methyl ester (L-NAME), a nonselective NOS inhibitor on blood pressure, plasma volume, and albumin escape during the early and delayed phases of endotoxin shock in conscious, chronically catheterized rats. Red blood cell volume and plasma volume were determined by using chromium-51-tagged erythrocytes and iodine-125-labeled albumin, respectively. Injection of lipopolysaccharide (LPS) 10 mg/kg i.v. resulted in a fall in blood pressure, hemoconcentration, and increased total-body albumin escape, which is reflected by a 25% reduction in plasma volume. When LPS was injected into animals pretreated with L-NAME (7.4 mumol/kg i.v. 15 minutes before LPS), losses in plasma volume and albumin escape were significantly greater than in rats that received LPS alone, despite that L NAME attenuated the hypotensive action of LPS. Aminoguanidine pretreatment (162 mumol/kg) had no effect on the early responses to LPS, whereas it was as potent as L-NAME in reversing hypotension when injected 70 minutes after LPS. Aminoguanidine treatment also prevented further losses in plasma volume and markedly attenuated total-body and organ albumin escape rates elicited by LPS. L NAME produced only a slight attenuation of LPS-induced losses in plasma volume and albumin escape in most organs studied, whereas it potentiated albumin extravasation in the lung. These results demonstrate that inhibition of cNOS potentiates, whereas inhibition of iNOS markedly attenuates, losses in plasma volume and albumin escape elicited by LPS, and suggest that selective inhibitors of iNOS may be more effective than nonselective inhibitors of all forms of NOS in the therapy of septic shock. PMID- 9351459 TI - Potentiation of the actions of bradykinin by angiotensin I-converting enzyme inhibitors. The role of expressed human bradykinin B2 receptors and angiotensin I converting enzyme in CHO cells. AB - Part of the beneficial effects of angiotensin I-converting enzyme (ACE) inhibitors are due to augmenting the actions of bradykinin (BK). We studied this effect of enalaprilat on the binding of [3H]BK to Chinese hamster ovary (CHO) cells stably transfected to express the human BK B2 receptor alone (CHO-3B) or in combination with ACE (CHO-15AB). In CHO-15AB cells, enalaprilat (1 mumol/L) increased the total number of low-affinity [3H]BK binding sites on the cells at 37 degrees C, but not at 4 degrees C, from 18.4 +/- 4.3 to 40.3 +/- 11.9 fmol/10(6) cells (P < .05; Kd, 2.3 +/- 0.8 and 5.9 +/- 1.3 nmol/L; n = 4). Enalaprilat preserved a portion of the receptors in high-affinity conformation (Kd, 0.17 +/- 0.08 nmol/L; 8.1 +/- 0.9 fmol/10(6) cells). Enalaprilat decreased the IC50 of [Hyp3-Tyr(Me)8]BK, the BK analogue more resistant to ACE, from 3.2 +/ 0.8 to 0.41 +/- 0.16 nmol/L (P < .05, n = 3). The biphasic displacement curve of the binding of [3H]BK also suggested the presence of high-affinity BK binding sites. Enalaprilat (5 nmol to 1 mumol/L) potentiated the release of [3H]arachidonic acid and the liberation of inositol 1,4,5-trisphosphate (IP3) induced by BK and [Hyp3-Tyr(Me)8]BK. Moreover, enalaprilat (1 mumol/L) completely and immediately restored the response of the B2 receptor, desensitized by the agonist (1 mumol/L [Hyp3-Tyr(Me)8]BK); this effect was blocked by the antagonist, HOE 140. Finally, enalaprilat, but not the prodrug enalapril, decreased internalization of the receptor from 70 +/- 9% to 45 +/- 9% (P < .05, n = 7). In CHO-3B cells, enalaprilat was ineffective. ACE inhibitors in the presence of both the B2 receptor and ACE enhance BK binding, protect high-affinity receptors, block receptor desensitization, and decrease internalization, thereby potentiating BK beyond blocking its hydrolysis. PMID- 9351460 TI - Role of tissue renin in the regulation of aldosterone biosynthesis in the adrenal cortex of nephrectomized rats. AB - The aim of the study was to investigate whether the adrenal renin-angiotensin system plays an independent role in the regulation of mineralocorticoid biosynthesis in the adrenal gland and to explore the mechanisms of this action. Twelve-week-old male Sprague-Dawley rats were studied: 22 rats were maintained on a regular diet; 27 and 22 rats received a low salt diet with and without treatment, respectively, with the angiotensin II (Ang II) AT1-subtype receptor antagonist losartan (10 mg/kg per day). A fraction of each group of rats underwent bilateral nephrectomy (n = 12, 15, and 10, respectively) and was killed 48 hours later. In an additional group of 24 (12 intact and 12 nephrectomized) rats, the effects of the Ang II AT2-subtype receptor antagonist PD123319 were investigated. In intact rats, plasma renin activity (PRA) and adrenal renin activity and expression were progressively raised by salt restriction and losartan, whereas aldosterone synthase mRNA and plasma aldosterone (PA) levels were increased by salt restriction and reduced by losartan. Forty-eight hours after nephrectomy, PRA fell to undetectable levels; in contrast, adrenal renin expression, assessed by semiquantitative reverse-transcriptase polymerase chain reaction (using GAPDH as a standard for gene expression), showed an 18-fold increase and was further increased after salt restriction and losartan (all P < .05). Also, adrenal renin activity was raised after nephrectomy and further increased after salt restriction (P < .05) and losartan. Cytochrome P450 aldosterone synthase expression in the adrenal cortex was stimulated by nephrectomy alone and by nephrectomy combined with low salt intake (P < .05), with consequent increases in PA concentrations. In losartan-treated salt restricted nephrectomized rats, cytochrome P450 aldosterone synthase expression (P < .05 versus nephrectomy alone and nephrectomy plus salt restriction) and PA concentrations were diminished (P < .05) in spite of the observed increases of adrenal renin expression. The AT2-receptor antagonism did not significantly affect PRA, adrenal renin, and aldosterone biosynthesis and production in either intact or nephrectomized salt-restricted rats. These results demonstrate that the adrenal renin-angiotensin system plays an independent role in the regulation of mineralocorticoid biosynthesis in vivo. This action is mediated primarily via the Ang II AT1-subtype receptors. PMID- 9351461 TI - Nitric oxide inhalation inhibits platelet aggregation and platelet-mediated pulmonary thrombosis in rats. AB - Endothelium-derived nitric oxide (NO) inhibits in vitro platelet aggregation via a cGMP-dependent mechanism. The effect of inhaled NO on platelet-mediated pulmonary thrombosis following intravenous thrombotic challenge with collagen was examined in rats and compared with the effect of G4120, a cyclic Arg-Gly-Asp containing synthetic pentapeptide that binds to the platelet glycoprotein IIb/IIIa receptor. Intraplatelet cGMP dose-dependently increased from 39 +/- 6 fmol/10(8) platelets in control to 46 +/- 6, 68 +/- 13, and 81 +/- 13 fmol/10(8) platelets after inhalation with 20, 40, and 80 ppm NO, respectively (P < .05 for 40 and 80 ppm). Ex vivo platelet aggregation of platelet-rich plasma induced by 1 microgram/mL collagen was reduced from 75 +/- 4% in control rats to 22 +/- 10% and 20 +/- 7% in rats ventilated with 40 and 80 ppm NO, respectively, and to 30 +/- 9% in G4120-treated rats (each P < .05 versus control). Circulating platelet counts 3 minutes after collagen injection were significantly higher in the inhaled NO and G4120 groups compared with control rats (250,000 +/- 18,000 and 223,000 +/- 10,000/microL versus 160,000 +/- 18,000/microL, each P < .05). The rise in pulmonary arterial pressure after collagen injection was significantly reduced in NO- and G4120-treated rats (26 +/- 1 and 27 +/- 1 versus 32 +/- 1 mm Hg in control rats, each P < .05). The number of pulmonary resistance vessels containing platelet thrombi was significantly smaller after inhaled NO and G4120 treatment compared with control (56 +/- 3% and 50 +/- 3% versus 68 +/- 3%, respectively; P < .05). Thus, NO inhalation reduces in vivo activation of circulating platelets and platelet-rich thrombosis in thromboembolic pulmonary hypertension. Inhalation of NO may be useful in cardiovascular diseases associated with platelet activation. PMID- 9351463 TI - Actions of cADP-ribose and its antagonists on contraction in guinea pig isolated ventricular myocytes. Influence of temperature. AB - Although it is becoming widely accepted that cADP-ribose (cADPR) can regulate calcium release from the endoplasmic reticulum in sea urchin eggs and in a variety of mammalian cell types, it remains controversial whether this substance might influence calcium release during excitation-contraction coupling in cardiac muscle. We have investigated possible actions of cADPR in intact cells isolated from guinea pig ventricle, paying particular attention to the possible influence of temperature. At 36 degrees C, myocyte contraction was influenced by cytosolic application of cADPR in a concentration-dependent manner (showing an approximately 30% increase in contraction with 5 mumol/L cADPR applied via a patch pipette in myocytes stimulated to fire action potentials at 1 Hz). Calcium transients measured with fura 2 were also increased by 5 mumol/L cADPR. Antagonists of cADPR reduced contraction at 36 degrees C (by approximately 35% with either 50 mumol/L 8-Br-cADPR or 5 mumol/L 8-amino-cADPR applied via the patch pipette). At room temperature (approximately 20 degrees C to 24 degrees C), no significant effects on contraction were detected with either cADPR or its antagonists. At 36 degrees C, treatment of the cells with a mixture of 2 mumol/L ryanodine and 1 mumol/L thapsigargin to suppress function of the sarcoplasmic reticulum stores of calcium prevented the action of 5 mumol/L cADPR applied via a patch pipette. These observations are consistent with an action of cytosolic cADPR to enhance calcium-induced calcium release from the sarcoplasmic reticulum in guinea pig ventricular myocytes at 36 degrees C. The observed influence of temperature under the conditions of our experiments is one factor that might help to account for failure to detect actions of cADPR and its analogues in some previous studies. PMID- 9351462 TI - Two isoforms of the mouse ether-a-go-go-related gene coassemble to form channels with properties similar to the rapidly activating component of the cardiac delayed rectifier K+ current. AB - HERG, the human ether-a-go-go-related gene, encodes a K(+)-selective channel with properties similar to the rapidly activating component of the delayed rectifier K+ current (IKr). Mutations of HERG cause the autosomal-dominant long-QT syndrome (LQTS), presumably by disrupting the normal function of IKr. The current produced by HERG is not identical to IKr, however, and the mechanism by which HERG mutations cause LQTS remains uncertain. To better define the role of Erg in the heart, we cloned Merg1 from mouse genomic and cardiac cDNA libraries. Merg1 has 16 exons and maps to mouse chromosome 5 in an area syntenic to human chromosome 7q, the map locus of HERG. We isolated three cardiac isoforms of Merg1: Merg1a is homologous to HERG and is expressed in heart, brain, and testes, Merg1a' lacks the first 59 amino acids of Merg1a and is not expressed abundantly, and Merg1b has a markedly shorter divergent N-terminal cytoplasmic domain and is expressed specifically in the heart. The Merg1 isoforms, like HERG, produce inwardly rectifying E-4031-sensitive currents when heterologously expressed in Xenopus oocytes. Merg1a and HERG produce currents with slow deactivation kinetics, whereas Merg1a' and Merg1b currents deactivate more rapidly. Merg1b coassembles with Merg1a to form channels with deactivation kinetics that are more rapid than those of Merg1a or HERG and nearly identical to IKr. In addition, a homologue of Merg1b is present in human cardiac and smooth muscle. Thus, we have identified a novel N-terminal Erg isoform that is expressed specifically in the heart, has rapid deactivation kinetics, and coassembles with the longer isoform in Xenopus oocytes. This N-terminal Erg isoform may determine the properties of IKr and contribute to the pathogenesis of LQTS. PMID- 9351464 TI - 17 beta-estradiol regulation of human endothelial cell basal nitric oxide release, independent of cytosolic Ca2+ mobilization. AB - Estradiol retards the development of atherosclerosis. Animal models have suggested that NO may be a critical effector molecule in this cardiovascular protection. In this study, female human umbilical vein endothelial cells (HUVECs) were propagated in phenol red-free gonadal hormone-free medium and pretreated with 17 beta-estradiol (E2). Reduced NO2- and NO3- (NOx) concentration, determined by chemiluminescence, demonstrated a rapid increase in basal HUVEC NO release in response to physiological concentrations of E2. The estrogen receptor (ER) antagonist ICI 164,384 inhibited the augmented NO release, demonstrating an ER-mediated component of this response. Because endothelial NO synthase (eNOS) activity is largely regulated by cytosolic Ca2+, relative [Ca2+]i in response to E2 was determined in a fluorometric assay. E2 did not promote HUVEC Ca2+ fluxes. Furthermore, eNOS activity in E2-pretreated endothelial whole-cell lysates was not dependent on additional Ca2+. Despite involving the ER, this is a nongenomic effect E2, as demonstrated by maintained responses in transcriptionally inhibited cells and by the rapidly (10 minutes) of cGMP formation in an NO bioassay. We demonstrate, for the first time, that independent of cytosolic Ca2+ mobilization, there is augmentation of eNOS activity with a resultant increase in HUVEC basal NO release in response to short-term estradiol exposure. Implications for the cardiovascular protective role of estrogen are discussed. PMID- 9351465 TI - [New therapeutic principle in the management of cardiac insufficiency: A II antagonists. Results of the ELITE study (Evaluation of Losartan In The Elderly)]. PMID- 9351466 TI - Deciphering protein sequence information through hydrophobic cluster analysis (HCA): current status and perspectives. AB - Ten years after the idea of hydrophobic cluster analysis (HCA) was conceived and first published, theoretical and practical experience has shown this unconventional method of protein sequence analysis to be particularly efficient and sensitive, especially with families of sequences sharing low levels of sequence identity. This extreme sensitivity has made it possible to predict the functions of genes whose sequence similarities are hardly if at all detectable by current one-dimensional (1D) methods alone, and offers a new way to explore the enormous amount of data generated by genome sequencing. HCA also provides original tools to understand fundamental features of protein stability and folding. Since the last review of HCA published in 1990 [1], significant improvements have been made and several new facets have been addressed. Here we wish to update and summarize this information. PMID- 9351467 TI - Propranolol-induced seizures in mice: the role of noradrenaline. AB - The effects of some noradrenergic agents, phenobarbitone, diazepam and phenytoin on seizures produced by propranolol were investigated in mice. Isoprenaline and DL-threo-3,4-dihydroxyphenylserine (DOPS) effectively antagonized the seizures elicited by propranolol. Pargyline and imipramine significantly attenuated propranolol-induced seizures and also significantly potentiated the protecting effect of DOPS against the seizures. alpha-Methyl-p-tyrosine, disulfiram and reserpine significantly potentiated propranolol-elicited seizures. However, DOPS significantly antagonized the seizure-potentiating effects of alpha-methyl-p tyrosine, disulfiram and reserpine. Phenylephrine, clonidine, prazosin, idazoxan, phenobarbitone, diazepam and phenytoin did not significantly alter propranolol induced seizures. These results suggest that propranolol-induced seizures in mice may involve a noradrenergic mechanism mediated via central beta-adrenoceptors. PMID- 9351468 TI - Drosophila unconventional myosin VI is involved in intra- and intercellular transport during oogenesis. AB - During mid-oogenesis of Drosophila, cytoplasmic particles are transported within the nurse cells and through ring canals (cytoplasmic bridges) into the oocyte by means of a microfilament-dependent mechanism. Video-intensified fluorescence timelapse microscopy, in combination with microinjections of antibodies directed against Drosophila 95F myosin, have revealed that this unconventional myosin of class VI is involved in the transport processes. The results indicate that certain cytoplasmic particles in the nurse cells move along microfilaments due to their direct association with myosin VI motors. Additional myosin-VI molecules located at the rim of the ring canals seem to be involved in particle transport into the oocyte. Microinjected mitochondria-specific dyes have revealed that some of these particles are mitochondria. PMID- 9351469 TI - Anti-granulocyte antibody suppression of active and passive anaphylactic shock in WBB6F1-W/Wv mice. AB - Our previous study revealed that anaphylactic shock can be produced in WBB6F1 W/Wv (abbreviated as W/Wv) mice. Because they are congenitally deficient in mast cells, we are certain that some other cell types are involved as mediator sources. In the present study, with the aim of examining the role of circulatory granulocytes, the effect of monoclonal antibody to a mouse granulocyte antigen, Gr-1, upon active and passive anaphylactic shock was tested in W/Wv mice using several mast cell-bearing strains as references. An intravenous injection of 40 micrograms of anti-granulocyte antibody one day before the antigen challenge produced a marked decrease in neutrophils and nearly complete abolition of lethal shock in W/Wv mice regardless of the sensitizing method. Both prevention of shock and reduction of neutrophils lasted for three days after the treatment with the antibody but not for six days. From these results, a reasonable conclusion would be that circulating Gr-1+ cells (predominantly composed of neutrophils) are the major mediator source. Reference experiments using mast cell-bearing strains revealed that the suppressive effect of anti-granulocyte antibody was also observed against active anaphylactic shock in C3H/HeN mice but not against active and passive anaphylactic shock in the other mice. PMID- 9351470 TI - Antimitotic effects of usnic acid on different biological systems. AB - Usnic acid is a biosynthesis product characteristic of several epiphytic lichens such as Evernia, Cladonia and Parmelia. Usnic acid has several interesting biological properties. It is an antibiotic and it also seems to exert an antimitotic action. It has even been postulated that usnic acid can play a role as an environmental indicator, since its concentration varies according to the presence of toxic agents. A series of tests have been run on different biological systems such as fungi, yeasts, plant cells and neoplastic human cell cultures in order to make a general evaluation of the properties of usnic acid and to highlight any analogy between its effects on phylogenetically distant organisms. The results obtained confirm some of the already known properties of usnic acid and identify concentration ranges that are active against cells from different organisms. Furthermore, at low concentrations, the acid displays a capacity to stimulate cell metabolism in some of the biological systems tested. PMID- 9351471 TI - Characterization of specific corticosterone binding sites in adrenal cortex plasma membrane and their localization by autoradiographic studies. AB - Specific corticosterone binding to calf adrenal cortex plasma membrane was measured using the biologically active radioligand [3H]corticosterone. Corticosterone binding was found to be time-dependent, saturable and reversible, and was reduced by more than 70% when membranes were pretreated with proteases. The population of corticosterone binding sites in calf adrenal cortex plasma membrane was homogeneous and displayed the following characteristics: equilibrium dissociation constant Kd = 77 +/- 8 nM and maximum specific binding capacity Bmax = 70,378 +/- 6,385 fmol/mg protein. The relative affinities of several structural analogues of steroids were deduced from competition assays. From these experiments we can conclude that the plasma membrane binding site characterized is selective for corticosterone and progesterone derivatives, and different from nuclear glucocorticoid, mineralocorticoid, estrogen and progestin receptors. Likewise, this corticosterone binding site is independent of mineralocorticoid and Na+, K(+)-ATPase digitalis receptors. From autoradiographic studies we suggest these corticosterone binding sites are located in the whole adrenal cortex. PMID- 9351472 TI - Recombinant synthesis of mouse Zn3-beta and Zn4-alpha metallothionein 1 domains and characterization of their cadmium(II) binding capacity. AB - Genetic engineering, coupled with spectroscopic analyses, has enabled the metal binding properties of the alpha and beta subunits of mouse metallothionein 1 (MT) to be characterized. A heterologous expression system in E.coli has led to high yields of their pure zinc-complexed forms. The cadmium(II) binding properties of recombinant Zn4-alpha MT and Zn3-beta MT have been studied by electronic absorption and circular dichroism. The former binds Cd(II) identically to alpha fragments obtained from mammalian organs, showing that the recombinant polypeptide behaves like the native protein. Titration of Zn3-beta MT with CdCl2 results in the formation of Cd3-beta MT. The addition of excess Cd(II) leads to Cd4-beta MT which, with the extra loading of Cd(II), unravels to give rise isodichroically to Cd9-beta MT. The effect of cadmium-displaced Zn(II) ions and excess Cd(II) above the full metal occupancy of three has been studied using Chelex-100. The Cd3-beta MT species is stable in the presence of this strong metal-chelating agent. PMID- 9351473 TI - Comparison of the rate of phagocytosis of orthorhombic cyclosporine A (CsA) and latex particles by alveolar macrophages from hamsters. AB - The aim of this study was to develop an in vitro model to estimate the clearance of pulmonary administered cyclosporine A (CsA). To do this we estimated the volume of CsA particles phagocytosed by alveolar macrophages (AM) lavaged from hamsters. AM were cultured with CsA particles at two doses of particles (0.1 mg or 0.5 mg) and at three incubation times (1 h, 6 h or 24 h). The AM were also incubated with or without latex particles. After incubation, AM were processed for light and electron microscopy and the mean volume of phagocytosed particles was estimated stereologically from micrographs of the cells. Here, however, the CsA particles were dissolved during the embedding process and only their negative images (vacuoles) could be detected. An indirect method was therefore developed. The volume of cytoplasmic vacuoles (called 'background' vacuoles) was estimated in control macrophages (without particles or with latex particles and subtracted from the total volume of vacuoles in macrophages incubated with CsA, which gave the volume of phagocytosed CsA. The volume of the 'background' vacuoles remained constant in all study conditions. At a dose of 0.1 mg CsA the volume phagocytosed per macrophage was 13.83 microns3 at 1 h, 8.43 microns3 at 6 h and 4.50 microns3 at 24 h. At a dose of 0.5 mg CsA, the volume phagocytosed varied from 26.59 microns3 at 1 h, to 4.13 microns3 at 6 h and 49.10 microns3 at 24 h. These results show no statistically significant dependence on time for either dose, and a statistically significant dose effect only at 24 h. With latex particles, the phagocytosed volume increased significantly with time and dose and was significantly higher than for CsA particles. This study showed that CsA particles are phagocytosed by AM from hamsters but to a lesser extent than latex particles. This difference could be correlated with physical properties, i.e. a difference between particle size and shape and/or chemical properties, latex particles being inert and CsA particles being peptidic. Moreover, different surface receptors on AM could be involved in the process of phagocytosis of CsA and latex particles. PMID- 9351474 TI - Continuous light exposure modifies the nocturnal increase in rat thymus type II thyroxine 5'-deiodinase. AB - In the present study we show that thymus type II thyroxine deiodinase activity exhibits a nyctohemeral profile, with basal values during the day and high values at night. This rhythmic character is dependent on neuroadrenergic input since exposure to continuous light at night completely abolished the nocturnal rise of the enzyme activity. However, treatment with isoproterenol under light exposure at night restored it. PMID- 9351475 TI - From apoplexy to stroke: plus ca change... PMID- 9351476 TI - From apoplexy to stroke. PMID- 9351477 TI - Reduced visual acuity in elderly people: the role of ergonomics and gerontechnology. AB - Gerontology is the scientific study of the ageing process and special problems of aged people. Ergonomics is an applied science for optimizing performance and productivity and reducing the risks of injury, discomfort and illness. Gerontechnology is concerned with fundamental and applied research on the complex interaction of elderly people with technological products and the built environment. It has the potential to improve the capability of people confronted by the challenges of ageing. We suggest that gerontechnology may have a particular role in relation to the reduction of visual acuity, and can improve the comfort and safety of older people. PMID- 9351478 TI - Prescription of antimicrobial agents to elderly people in relation to the type of infection. AB - OBJECTIVE: to describe how frequently antimicrobial agents are prescribed for elderly people and to examine the prescribing practice of treating physicians. DESIGN: for each of the 1196 subjects examined, a review of the medical records was carried out for the year preceding the examination and data on antimicrobial prescriptions, including types of infections, were recorded. The sales statistics of antimicrobial agents in this population were compared with the nation-wide ones, collected by the National Agency for Medicines. SETTING: a health centre in Lieto, a rural district in southwestern Finland; 1990-91. SUBJECTS: 1196 subjects, aged 64-97 years, 488 men and 708 women. MAIN OUTCOME MEASURES: frequencies of prescriptions of specified antimicrobial agents, including types of infection in relation to gender and form of care over a 1-year period. RESULTS: more women (36%) than men (28%) had received antimicrobial agents. The proportion who had received such agents increased with increasing age, the trend being more marked in men. The mean number of prescriptions per user per year was slightly lower in men than women (0.6 and 0.7 respectively). In both sexes, 54% of the users of antimicrobial agents had received only one prescription. Cephalosporins and penicillins were the most commonly prescribed agents. Among those who had received three or more antimicrobial prescriptions, cephalosporins had been used most frequently. Elderly people living in long-term institutional care were treated with antimicrobial agents more frequently than those living outside institutions. Of all antimicrobial prescriptions given to women, 60% were prescribed for urinary tract infections, 21% for respiratory infections and 8% for skin infections. The figures for men were 18, 45 and 10%. CONCLUSIONS: multiple use of antimicrobial agents is common in old age, especially in those in institutions. More attention should be to the provision of appropriate antimicrobial treatment. PMID- 9351479 TI - The Parkinson's Disease Questionnaire (PDQ-39): development and validation of a Parkinson's disease summary index score. AB - OBJECTIVES: to briefly outline the development and validation of the Parkinson's Disease Questionnaire (PDQ-39) and then to provide evidence for the use of the measure as either a profile of health status scores or a single index figure. DESIGN: the PDQ-39 was administered in two surveys: a postal survey of patients registered with local branches of the Parkinson's Disease Society of Great Britain (n = 405) and a survey of patients attending neurology clinics for treatment for Parkinson's disease (n = 146). Data from the eight dimensions of the PDQ-39 were factor-analysed. This produced a single factor on the data from both surveys. OUTCOME MEASURES: the eight dimensions of the PDQ-39 and the new single index score-the Parkinson's disease summary index (PDSI), together with clinical assessments (the Columbia rating scale and the Hoehn and Yahr staging score). RESULTS: in the postal survey 227 patients returned questionnaires (58.2%). AH 146 patients approached in the clinic sample agreed to take part. Higher-order principal-components factor analysis was undertaken on the eight dimensions of the PDQ-39 and produced one factor on both datasets. Consequently it was decided that the scores of the eight domains could be summed to produce a single index figure. The psychometric properties of this index were explored using reliability tests and tests of construct validity. The newly derived single index was found to be both internally reliable and valid. DISCUSSION: data from the PDQ-39 can be presented either in profile form or as a single index figure. The profile should be of value in studies aimed at determining the impact of treatment regimes upon particular aspects of functioning and well-being in patients with Parkinson's disease, while the PDSI will provide a summary score of the impact of the illness on functioning and well-being and will be of use in the evaluation of the overall effect of different treatments. Furthermore, the PDSI reduces the number of statistical comparisons and hence the role of chance when exploring data from the PDQ-39. PMID- 9351480 TI - Effect of intermittent cyclical disodium etidronate therapy on bone mineral density in men with vertebral fractures. AB - OBJECTIVES: to investigate the effects of oral intermittent cyclical etidronate therapy on bone mineral density (BMD) in men with idiopathic vertebral osteoporosis. DESIGN: consecutive case series. SETTING: regional specialist clinic for metabolic bone disease. SUBJECTS: 42 men aged 35-81 (median 60.5) with established vertebral crush fractures and back pain, in whom secondary causes of osteoporosis had been excluded. INTERVENTION: repeated cycles of treatment with oral disodium etidronate 400 mg daily for 14 days followed by oral calcium 500 mg as citrate daily for 76 days. OUTCOME MEASURES: BMD measurement of the lumbar spine and femoral neck by dual energy x-ray absorptiometry at 6-12-month intervals; bone biochemistry (serum calcium, phosphate, alkaline phosphatase and urine calcium/creatinine and hydroxyproline/creatinine ratios) at 6-month intervals. RESULTS: all 42 men have been treated for more than 18 months, and 35 of them for more than 24 months. Median follow-up for the group as a whole is 31 months (range 18-45). The treatment was well tolerated. BMD at the lumbar spine increased by a mean of 0.024 g/cm2 per year of follow-up (95% confidence interval 0.017-0.032 g/cm2). This is equivalent to an average annual rate of change of 3.2% of baseline values. There was a small, non-significant rise in mean BMD at the hip equivalent to 0.7% of baseline values per year. Serum alkaline phosphatase tended to fall in the first 6 months of treatment, returning to baseline values at 2 years. Serum calcium and phosphate were unchanged and no decrease in urinary calcium/creatinine ratio or hydroxyproline/creatinine ratio was seen. CONCLUSIONS: intermittent cyclical etidronate therapy increased lumbar spine BMD over a 2-year period in an unselected group of men with osteoporotic vertebral fractures. This treatment warrants further evaluation in a randomized controlled trial. PMID- 9351482 TI - Drug-related problems in elderly patients admitted to Tayside hospitals, methods for prevention and subsequent reassessment. AB - INTRODUCTION: although drug-related problems (DRPs) are known to be prevalent in elderly patients, the literature on prevention of iatrogenic disease is sparse. The present study addresses this requirement. OBJECTIVES: to assess the incidence of DRPs in elderly patients admitted to Tayside hospitals before (phase I) and after (phase II) implementation of preventive strategies. DESIGN: all elderly people admitted to hospital were screened by a pharmacist; individual case reviews were prepared for all those with a potential DRP and reviewed by a three member panel which made a final decision on the presence of a DRP and its contribution to admission. SETTING: all hospital wards admitting elderly patients in the Tayside region of Scotland. SUBJECTS: 1011 elderly patient admissions over a 9-month period (phase I); 857 elderly patient admissions over an 8-month period (phase II). MAIN OUTCOME MEASURES: incidence of DRPs before and after targeted intervention strategies (information bulletin for general practitioners, patient information leaflet, oral presentation to trainee general practitioners). RESULTS: in phase I, the incidence of DRPs was 144/1011 (14.2%), with 54/1011 (5.3%) of the admissions identified as being definitely or probably drug-related. Non-steroidal anti-inflammatory drugs (NSAIDs) were the main drug group involved, being responsible for 15/54 (28%) of admissions primarily due to a DRP. Over 66% of admissions due to adverse effects of NSAIDs were considered to be definitely preventable. In phase II, after targeted intervention strategies, there was no significant reduction in total incidence of DRPs or incidence of DRPs related to NSAIDs. However, there appeared to be an improvement in the first 4 months, and a significant drop in NSAID prescribing in Tayside compared with the rest of Scotland was observed. CONCLUSION: DRPs remain a significant problem in elderly patients and NSAIDs are the major contributor. The intervention strategies used in the study were not demonstrably effective, but a continuous programme of education may be necessary to limit NSAID use. PMID- 9351483 TI - Drug use by demented and non-demented elderly people. AB - AIM: to determine the use of drugs by demented and non-demented elderly people in a population, by dementia status and type, age, sex and accommodation type. METHOD: data were obtained from the Kungsholmen project, a longitudinal community study of people over 75 in Stockholm, Sweden. RESULTS: 85% used at least one medicinal drug, and of these 12% were demented. Mean numbers of drugs used were 2.8 for demented and 3.2 for non-demented people. 45% of demented people and 38% of non-demented people used psychotropic agents. Psychotropic use was higher in women and increased with institutionalization. Antipsychotic agents were used more by demented (22%) than by non-demented (3.5%) people: this was largely explained by differences in accommodation type. The odds ratio (OR) for use of antipsychotics by those in institutions compared with those living in their own homes was 9.32. Opioids were commonly prescribed for demented people. The proportions taking opioids in those using analgesics were 42% in demented and 23% in non-demented people (OR 2.07). Laxatives were used by 18% of the demented people in institutions compared with 39% of non-demented people in institutions. CONCLUSION: being in an institution had a stronger association with the use of certain drugs (e.g. psychotropics) than did dementia status. Demented people, especially those in institutions, used a large number of antipsychotics and opioids, but fewer laxatives and minor analgesics. Prescribers and institutional staff should be aware of these factors so they can optimize patient treatment. PMID- 9351481 TI - Medically recognized urinary incontinence and risks of hospitalization, nursing home admission and mortality. AB - OBJECTIVES: this study examined the association between medically recognized urinary incontinence and risk of several disease conditions, hospitalization, nursing home admission and mortality. DESIGN: review and abstraction of medical records and computerized data bases from 5986 members, aged 65 years and older, of a large health maintenance organization in northern California. RESULTS: there was an increased risk of newly recognized urinary incontinence following a diagnosis of Parkinson's disease, dementia, stroke, depression and congestive heart failure in both men and women, after adjustment for age and cohort. The risk of hospitalization was 30% higher in women following the diagnosis of incontinence [relative risk (RR) = 1.3, 95% confidence interval (CI) = 1.2-1.5] and 50% higher in men (RR = 1.5, 95% CI = 1.3-1.6) after adjustment for age, cohort and co-morbid conditions. The adjusted risk of admission to a nursing facility was 2.0 times greater for incontinent women (95% CI = 1.7-2.4) and 3.2 times greater for incontinent men (95% CI = 2.7-3.8). In contrast, the adjusted risk of mortality was only slightly greater for women (RR = 1.1; 95% CI = 0.99 1.3) and men (RR= 1.2; 95% CI= 1.1-1.4). CONCLUSIONS: urinary incontinence increases the risk of hospitalization and substantially increases the risk of admission to a nursing home, independently of age, gender and the presence of other disease conditions, but has little effect on total mortality. PMID- 9351484 TI - Functional assessment scales in detecting dementia. AB - AIM: to evaluate the use of different functional scales in detecting dementia in a population study. METHODS: the study is part of the Helsinki Ageing Study. A random sample of 795 subjects aged 75 (n = 274), 80 (n = 266) and 85 years (n = 255) was taken. The prevalences of dementia (DSM-III-R criteria) in these age groups were 4.6, 13.1 and 26.7% respectively. The functional scale scores were known for 71% of the non-demented and 66% of the demented subjects. A structured questionnaire completed by a close informant included four functional scales: the index of activities of daily living (ADL), the modified Blessed dementia scale (DS), the instrumental activities of daily living scale (IADL) and the Functional Assessment Questionnaire (FAQ). RESULTS: all the functional scales discriminated demented from non-demented subjects. Based on receiver operating characteristics analysis, the area under the curve (95% confidence interval) was 0.90 (0.80-0.94) for the ADL, 0.94 (0.87-0.97) for the DS, 0.95 (0.90-0.98) for the IADL and 0.96 (0.92-0.98) for the FAQ. The effects of age, sex and education in detecting dementia were minor or non-existent in the ADL, DS and FAQ scales, but age had an effect on the performance of the IADL scale. All the scales detected even mild dementia adequately. CONCLUSIONS: functional scales can be used in detecting dementia when functional assessment is already used for other purposes, such as among elderly primary care patients. PMID- 9351485 TI - Death certification in treated cases of presenile Alzheimer's disease and vascular dementia in Scotland. AB - INTRODUCTION: although death certification data are commonly used in dementia epidemiology, their reliability has been questioned. METHODS: death certificates were available from the Registrar General for Scotland for all patients with Alzheimer's disease/presenile dementia (AD PSD) or vascular dementia (VaD) who had died in Scotland up until 31 December 1994. Primary (immediate and underlying) and contributory causes of death were noted as well as place of death. Occupations of male patients were obtained from death certificates or from case notes and classified according to the Standard Occupational Classification. Bronchopneumonia was considered a non-specific cause of death and specific causes of death were classified as: cardiac disease, dementia, cerebrovascular disease, neoplasms, other vascular diseases and other diseases. Place of death was recorded as psychiatric hospital, district general hospital, nursing home or private residence. RESULTS: death certificates of 398 people who had been treated for AD PSD and 348 who had been treated for VaD were identified. Bronchopneumonia was the most common immediate cause of death in the AD PSD group (70.9%) but less so for the VaD group (51.7%). For both groups place of death was associated with significant differences in pneumonia being reported as the immediate cause of death as well as specific underlying and contributory causes of death. Dementia was recorded for 90.5% of AD PSD patients but for only 49.7% of the VaD group. CONCLUSIONS: Scottish death certificate data significantly underestimate the prevalence of presenile VaD. Changes in patterns of institutional care may affect dementia rates estimated from death certificate data. PMID- 9351486 TI - Alzheimer's disease: clinical predictors of rates of intellectual decline--a prospective study. PMID- 9351487 TI - Factor V Leiden in a healthy Northern Ireland elderly population. PMID- 9351488 TI - Predictors of relapse in elderly diabetic patients admitted with sulphonylurea induced severe hypoglycaemic attacks. PMID- 9351489 TI - Can response to partial sleep deprivation in depressed patients be predicted by regional changes of cerebral blood flow? AB - The possible predictive value of regional cerebral perfusion patterns with respect to the response to partial sleep deprivation (PSD) was evaluated in 15 major depressive patients (mean age = 54.9 years, mean Hamilton depression score = 21.6). Patients were studied with single photon emission computed tomography with technetium-99 m-D,L-hexamethyl-propylene amine oxime. Scans were performed on the morning before and after (at 08.00 h) PSD. Responders to PSD had significantly higher perfusion in the right orbitofrontal cortex than did non responders before PSD. Multiple regression analysis indicated that right orbitofrontal/basal cingulate perfusion (r = -0.77, P < 0.001) before PSD, and left inferior temporal perfusion (r = 0.59, P = 0.01) after PSD, were fairly accurate predictors of change in Hamilton depression scores. Thus, it appears that the orbitofrontal cortex and the cingulate are involved in PSD and may serve as predictors of therapeutic response. PMID- 9351490 TI - Is working memory intact in alcoholics? An ERP study. AB - Few investigators have applied the working memory theory to studies on abstinent chronic alcoholics, though it has been reported that the deficits in short-term memory appear to be specific to visuo-spatial and problem-solving tasks. In the present study, we recorded ERPs from 40 male control subjects and 78 alcoholics performing a modified delayed matching to sample task. To minimize the possible confound of retinotopic projections for the matching stimuli, in contrast to the non-matching stimuli, we employed a unique set of stimuli in our delayed matching to sample task. Our results indicate that an ERP component, occurring at approximately 250 ms post-stimulus, may be a reflection of the ERP mnemonic effect for working memory. This component distinguishes the two groups at the right occipitotemporal region, providing evidence of right hemisphere dysfunction in alcoholics. Thus, the current experiment may show electrophysiological evidence of working memory deficits in alcoholics. PMID- 9351491 TI - Extrapyramidal side effects with risperidone and haloperidol at comparable D2 receptor occupancy levels. AB - Risperidone is an antipsychotic drug with high affinity at dopamine D2 and serotonin 5-HT2 receptors. Previous clinical studies have proposed that risperidone's pharmacologic profile may produce improved efficacy for negative psychotic symptoms and decreased propensity for extrapyramidal side effects; features shared by so-called 'atypical' neuroleptics. To determine if routine risperidone treatment is associated with a unique degree of D2 receptor occupancy and pattern of clinical effects, we used [123I]IBZM SPECT to determine D2 occupancy in subjects treated with routine clinical doses of risperidone (n = 12) or haloperidol (n = 7). Both risperidone and haloperidol produced D2 occupancy levels between approximately 60 and 90% at standard clinical doses. There was no significant difference between occupancy levels obtained with haloperidol or risperidone. Drug-induced parkinsonism was observed in subjects treated with risperidone (42%) and haloperidol (29%) and was observed at occupancy levels above 60%. Based on these observations, it is concluded that 5-HT2 blockade obtained with risperidone at D2 occupancy rates of 60% and above does not appear to protect against the risk for extrapyramidal side effects. PMID- 9351492 TI - [123I]IBZM SPECT in patients treated with typical and atypical neuroleptics: relationship to drug plasma levels and extrapyramidal side effects. AB - [123I]Iodobenzamide (IBZM) is an iodine-labeled dopamine receptor ligand and can be used to visualize brain D2 receptors in humans with single photon emission computerized tomography (SPECT). The ratio of striatal IBZM uptake to uptake in frontal cortex (ST/FC ratio) represents a semiquantitative measure of D2 receptor binding in the striatum. Our study sample included six patients treated with haloperidol (3.0-8.0 mg/day orally; one patient with an average of 0.9 mg/day intramuscularly), five patients with benperidol (9.0-15.0 mg/day orally) and nine patients treated with clozapine (200.0-600.0 mg/day orally). Typical neuroleptics (TNs) and atypical neuroleptics (ANs) were significantly different in their ST/FC ratios. The ST/FC ratios indicated that patients treated with benperidol exhibited the lowest ST/FC ratios, with increasingly higher ratios in patients on haloperidol or clozapine. We found a curvilinear relationship between the ST/FC ratios and the dose/kg body wt. of TNs and ANs on the basis of a dose normalization according to Ki-values of the neuroleptic at D2 receptors and a weaker, but also curvilinear relationship between ST/FC ratios and normalized dosages according to clinically defined chlorpromazine equivalents. The specific uptake of IBZM did not correlate with the plasma levels of the TN haloperidol at the present dose range (0-12.4 ng/ml). For clozapine, a meaningful negative correlation between plasma levels and ST/FC ratio could be established. There was a negative continuous correlation between uptake of IBZM and extrapyramidal side effects, which is different from the threshold-based relationship between extrapyramidal side effects and IBZM uptake reported previously. PMID- 9351493 TI - Gender differences in D2 dopamine receptor binding in drug-naive patients with schizophrenia: an [123I]iodobenzamide single photon emission computed tomography study. AB - Recent studies have described a left lateralized striatal asymmetry of D2 dopamine receptors in male patients with schizophrenia. To replicate this finding and to explore its potential functional consequences, we investigated the D2 dopamine receptor system in 23 drug-naive patients with schizophrenia using single photon emission computed tomography (SPECT). Patients were examined in the drug-naive state and 72 h after completing a standardized neuroleptic treatment with benperidol (12-16 mg/day) for 25 days. Each SPECT examination comprised two scans: the first scan was taken 2 h after intravenous injection of 185 MBq [123I]iodobenzamide. After completion of the first scan, patients received benperidol (8 mg) intravenously. The second scan was started 20 min later. For analysis, basal ganglia to frontal cortex ratios were calculated. Fifteen of the 23 patients originally recruited completed the study on day 28. When compared to female patients, male patients showed a left lateralized asymmetry of striatal D2 dopamine receptor binding in the drug-naive state with an almost significant (P = 0.07) sex x hemisphere interaction. In the male patients, benperidol challenge led to a reversal of asymmetry patterns. These findings support previous reports of a left lateralized striatal D2 receptor binding in drug-naive male patients with schizophrenia and suggest that this asymmetry may affect the binding of conventional neuroleptics such as benperidol at the D2 dopamine receptor. PMID- 9351494 TI - Effects of clozapine on rat striatal muscarinic receptors coupled to inhibition of adenylyl cyclase activity and on the human cloned m4 receptor. AB - 1. Clozapine has recently been claimed to behave as a selective and full agonist at the cloned m4 muscarinic receptor artificially expressed in Chinese hamster ovary (CHO) cells. In the present study we have investigated whether clozapine could activate the rat striatal muscarinic receptors coupled to the inhibition of adenylyl cyclase activity, considered as pharmacologically equivalent to the m4 gene product. In addition, we have examined the effect of the drug on various functional responses following the activation of the cloned m4 receptor expressed in CHO cells. 2. In rat striatum, clozapine (1 nM-10 microM) caused a slight inhibition of forskolin-stimulated adenylyl cyclase activity, which was not counteracted by 10 microM atropine. On the other hand, clozapine antagonized the inhibitory effect of acetylcholine with a pA2 value of 7.51. Moreover, clozapine (1 microM) failed to inhibit dopamine D1 receptor stimulation of adenylyl cyclase activity, but counteracted the inhibitory effect of carbachol (CCh). Clozapine displaced [3H]-N-methylscopolamine ([3H]-NMS) bound to striatal M4 receptors with a monophasic inhibitory curve and a pKi value of 7.69. The clozapine inhibition was not affected by the addition of guanosine-5'-O-(thio)triphosphate (GTPgammaS). 3. In intact CHO cells, clozapine inhibited forskolin-stimulated cyclic AMP accumulation with an EC50 of 31 nM. This effect was antagonized by atropine. CCh produced a biphasic effect on cyclic AMP levels, inhibiting at concentrations up to 1 microM (EC50=50 nM) and stimulating at higher concentrations (EC50 = 7 microM). Clozapine (0.3-5 microM) antagonized the CCh stimulation of cyclic AMP with a pKi value of 7.47. Similar results were obtained when the adenylyl cyclase activity was assayed in CHO cell membranes. 4. In CHO cells pretreated with the receptor alkylating agent 1-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline (10 microM), the maximal inhibitory effect of clozapine on cyclic AMP formation was markedly reduced, whereas the CCh inhibitory curve was shifted to the right with no change in the maximum. 5. As in rat striatum, in CHO cell membranes the displacement of [3H]-NMS binding by clozapine yielded a monophasic curve which was not affected by GTPgammaS. 6. Clozapine (10 nM-10 microM) had a small stimulant effect (approximately 20%) on the binding of [35S] GTPgammaS to CHO cell membranes, whereas CCh caused a 250% increase of radioligand binding. Moreover, clozapine (50 nM-5 microM) antagonized the CCh stimulated [35S]-GTPgammaS binding with a pA2 value of 7.48. 7. These results show that at the striatal M4 receptors clozapine is a potent and competitive antagonist, whereas at the cloned m4 receptor it elicits both agonist and antagonist effects. Thus, clozapine behaves as a partial agonist, rather than as a full agonist, at the m4 receptor subtype, with intrinsic activity changing as a function of the coupling efficiency of the receptor to effector molecules. PMID- 9351495 TI - Relationships between structure and vascular activity in a series of benzylisoquinolines. AB - 1. In the present work, the properties of 3-methyl isoquinoline, 3,4 dihydropapaverine, tetrahydropapaverine and tetrahydropapaveroline were compared with those of papaverine and laudanosine. The work includes functional studies on rat isolated aorta contracted with noradrenaline, caffeine or KCl, and a determination of the affinity of the compounds for alpha1-adrenoceptors and calcium channel binding sites, with [3H]-prazosin, [3H]-nitrendipine and [3H]-(+) cis-diltiazem binding to rat cerebral cortical membranes. The effects of papaverine derivatives on the different molecular forms of cyclic nucleotide phosphodiesterases (PDE) isolated from bovine aorta were also determined. 2. The three papaverine derivatives show greater affinity than papaverine for the [3H] prazosin binding site. They are therefore more selective as inhibitors of [3H] prazosin binding as opposed to [3H]-(+)-cis-diltiazem, while papaverine appears to have approximately equal affinity for both. [3H]-nitrendipine binding was not affected by either papaverine or papaverine derivatives in concentrations up to 100 microM. 3-Methylisoquinoline had no effect on any of the binding sites assayed. 3. Contractions evoked by noradrenaline (1 microM) in rat aorta were inhibited in a concentration-dependent manner by 3,4-dihydropapaverine, tetrahydropapaverine and with a lower potency, by tetrahydropapaveroline. In Ca2+ free solution, tetrahydropapaverine and to a lesser extent, tetrahydropapaveroline, inhibited the noradrenaline (1 microM) evoked contraction in a concentration-dependent manner and did not modify the phasic contractile response evoked by caffeine (10 mM). This suggests that these alkaloids do not act at the intracellular level, unlike papaverine which inhibits the contractile response to caffeine and noradrenaline. 4. Inositol phosphates formation induced by noradrenaline (1 microM) in rat aorta was inhibited by tetrahydropapaverine (100 microM) and tetrahydropapaveroline (300 microM), thus suggesting that alpha1D-adrenoceptors are coupled to phosphoinositide metabolism in rat aorta. 5. Unlike papaverine, which has a significant effect on all the PDE isoforms, the three alkaloids assayed did not have an inhibitory effect on the different forms of PDE isolated from bovine aorta. 6. These results provide evidence that papaverine derivatives with a partially or totally reduced isoquinoline ring have a greater affinity for alpha1-adrenoceptors and a lower affinity for benzothiazepine sites in the Ca2+-channel than papaverine. This structural feature also implies a loss of the inhibitory activity on PDE isoforms. The planarity of the isoquinoline ring (papaverine) impairs the interaction with the alpha1-adrenoceptor site and facilitates it with the Ca2+-channels and PDEs, whereas the more flexible tetrahydroisoquinoline ring increases the binding to alpha1-adrenoceptors. PMID- 9351496 TI - Neurokinin A-LI release after antigen challenge in guinea-pig bronchial tubes: influence of histamine and bradykinin. AB - 1. Our aim was to determine if antigen challenge stimulates sensory nerves and provokes the release of tachykinins. The involvement of histamine and bradykinin was studied by using specific receptor antagonists. Capsaicin-induced responses were also examined. Experiments were performed in vitro on tracheal and bronchial preparations from ovalbumin-sensitized guinea-pigs. 2. Characterization of ovalbumin-induced contraction, with regard to histamine and bradykinin, was carried out on airway ring preparations in the presence of phosphoramidon. The histamine H1 receptor antagonist pyrilamine reduced allergen-induced bronchial contractions by about 30%, whereas the bradykinin B2 receptor antagonist icatibant (Hoe 140) did not significantly affect the response. Combined treatment with pyrilamine (1 microM) and icatibant (0.1 microM) reduced the contractions by about 80%, indicating a synergistic inhibitory action. Tracheal preparations were not significantly affected by treatments, neither were capsaicin-induced contractions. 3. To study the outflow of tachykinins, we used a perfused bronchial-tube preparation, allowing simultaneous measurement of smooth muscle tension and mediator release. Neurokinin A-like immunoreactivity (NKA-LI) and substance P-like immunoreactivity (SP-LI) were determined by radioimmunoassay. 4. The results of the perfusion study showed an increased outflow of NKA-LI into the perfusate in response to ovalbumin (127% of basal) challenge. SP-LI determined in some of the samples showed a much lower amount (40 to 70 times lower) of SP-LI than NKA-LI. Treatment with icatibant and pyrilamine, separately and in combination, significantly reduced the ovalbumin-induced NKA-LI outflow by 38%, 26% and 22%, respectively. 5. Capsaicin-induced outflow (124% of basal) was not significantly affected by treatments (icatibant 121%, pyrilamine 107% and combined treatment 111% of basal). However, when pyrilamine was present the increased outflow was not statistically significant. 6. In conclusion, we found that allergen provocation of guinea-pig bronchi caused an increased outflow of NKA-LI that was reduced by treatment with both pyrilamine and icatibant. These findings demonstrate that the allergen-induced release of histamine and bradykinin stimulate sensory nerves and thereby increase outflow of tachykinins that contribute to the allergic reaction. PMID- 9351497 TI - Role of ATP in fast excitatory synaptic potentials in locus coeruleus neurones of the rat. AB - 1. Intracellular recordings were made in a pontine slice preparation of the rat brain containing the nucleus locus coeruleus (LC). The pressure application of alpha,beta-methylene ATP (alpha,beta-meATP) caused reproducible depolarizations which were depressed by suramin (30 microM) and abolished by suramin (100 microM). Pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS; 10, 30 microM) also concentration-dependently inhibited the alpha,beta-meATP-induced depolarization, although with a much slower time-course than suramin. Almost complete inhibition developed with 30 microM PPADS. Reactive blue 2 (30 microM) did not alter the effect of alpha,beta-meATP, while reactive blue 2 (100 microM) slightly depressed it. 2. Pressure-applied (S)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) also depolarized LC neurones. Kynurenic acid (500 microM) depressed and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 50 microM) abolished the response to AMPA. Suramin (100 microM) potentiated the AMPA effect. 3. Pressure-applied noradrenaline hyperpolarized LC neurones. Suramin (100 microM) did not alter the effect of noradrenaline. 4. Focal electrical stimulation evoked biphasic synaptic potentials consisting of a fast depolarization (p.s.p.) followed by a slow hyperpolarization (i.p.s.p.). A mixture of D(-)-2-amino-5-phosphonopentanoic acid (AP-5; 50 microM), CNQX (50 microM) and picrotoxin (100 microM) depressed both the p.s.p. and the i.p.s.p. Under these conditions suramin (100 microM) markedly inhibited the p.s.p., but did not alter the i.p.s.p. In the combined presence of AP-5 (50 microM), CNQX (50 microM), picrotoxin (100 microM), strychnine (0.1 microM), tropisetron (0.5 microM) and hexamethonium (100 microM), a high concentration of suramin (300 microM) almost abolished the p.s.p. without changing the i.p.s.p. 5. In the presence of kynurenic acid (500 microM) and picrotoxin (100 microM), PPADS (30 microM) depressed the p.s.p. Moreover, the application of suramin (100 microM) to the PPADS (30 microM)-containing medium failed to cause any further inhibition. Neither PPADS (30 microM) nor suramin (100 microM) altered the i.p.s.p. 6. It was concluded that the cell somata of LC neurones are endowed with excitatory P2 purinoceptors. ATP may be released either as the sole transmitter from purinergic neurones terminating at the LC or as a co-transmitter of noradrenaline from recurrent axon collaterals or dendrites of the LC neurones themselves. PMID- 9351498 TI - Potentiation by vasopressin of adrenergic vasoconstriction in the rat isolated mesenteric artery. AB - 1. The aim of the present study was to investigate in rat mesenteric artery rings whether low concentrations of vasopressin could modify the contractile responses to noradrenaline and electrical stimulation of perivascular nerves. 2. Vasopressin (10[10]-10[-7] M) caused concentration-dependent contractions (pD2 = 8.36+/-0.09). The V1-receptor antagonist d(CH2)5Tyr(Me)AVP (10[-9]-10[-8] M) produced parallel rightward shifts of the control curve for vasopressin. Schild analysis yielded a pA2 value of 9.83 with a slope of 1.10+/-0.14. 3. Vasopressin (3 x 10[-10] and 10[-9] M) caused concentration-dependent potentiation of the contractions elicited by electrical stimulation (2-8 Hz; 0.2 ms duration for 30 s) and produced leftward shifts of the concentration-response curve for noradrenaline. The V1-receptor antagonist induced concentration-dependent inhibitions of potentiation induced by vasopressin. The selective V1-receptor agonist [Phe2, Orn8]-vasotocin (3 x 10[10] and 10[-9] M) induced potentiation of electrical stimulation-evoked responses which was also inhibited in the presence of the V1 antagonist (10[-8] M). In contrast, the V2-receptor agonist deamino-8-D arginine vasopressin (desmopressin 10[-8]-10[-7] M) did not modify the electrical stimulation-induced responses and the V2-receptor antagonist [d(CH2)5, D-Ile2, Ile4, Arg8]-vasopressin (10[-8]-10[-7] M) did not affect the potentiation evoked by vasopressin. 4. In artery rings contracted by 10(-6) M noradrenaline in the presence of 10(-6) M guanethidine and 10(-6) M atropine, electrical stimulation (2, 4 and 8 Hz) produced frequency-dependent relaxations which were unaffected by 10(-9) M vasopressin but abolished by 10(-6) M tetrodotoxin. 5. Vasopressin also potentiated contractions elicited by KCl and contractions induced by addition of CaCl2 to KCl depolarized vessels. The augmenting effects were inhibited by the V1 antagonist. 6. In the presence of the calcium antagonist nifedipine (10[-6] M), vasopressin failed to enhance the contractile responses to electrical stimulation, noradrenaline and KCl. 7. The results demonstrate that low concentrations of vasopressin strongly potentiate the contractions to adrenergic stimulation and KCl depolarization. This effect appears to be mediated by V1 receptor stimulation which brings about an increase in calcium entry through dihydropyridine-sensitive calcium channels. PMID- 9351499 TI - RPR 106541, a novel, airways-selective glucocorticoid: effects against antigen induced CD4+ T lymphocyte accumulation and cytokine gene expression in the Brown Norway rat lung. AB - 1. The effects of a novel 17-thiosteroid, RPR 106541, were investigated in a rat model of allergic airway inflammation. 2. In sensitized Brown Norway rats, challenge with inhaled antigen (ovalbumin) caused an influx of eosinophils and neutrophils into the lung tissue and airway lumen. In the lung tissue there was also an accumulation of CD4+ T lymphocytes and increased expression of mRNA for interleukin-4 (IL-4) and IL-5, but not interferon-gamma (IFN-gamma). These findings are consistent with an eosinophilia orchestrated by activated Th2-type cells. 3. RPR 106541 (10-300 microg kg[-1]), administered by intratracheal instillation into the airways 24 h and 1 h before antigen challenge, dose dependently inhibited cell influx into the airway lumen. RPR 106541 (100 microg kg[-1]) caused a significant (P<0.01) (98%) inhibition of eosinophil influx and a significant (P<0.01) (100%) inhibition of neutrophil influx. RPR 106541 was approximately 7 times and 4 times more potent than budesonide and fluticasone propionate, respectively. 4. When tested at a single dose (300 microg kg[-1]), RPR 106541 and fluticasone each caused a significant (P<0.01) (100%) inhibition of CD4+ T cell accumulation in lung tissue. Budesonide (300 microg kg[-1]) had no significant effect. RPR 106541 and fluticasone (300 microg kg[-1]), but not budesonide (300 microg kg[-1]), significantly (P<0.05) inhibited the expression within lung tissue of mRNA for IL-4. RPR 106541 (300 microg kg[-1]) also significantly (P<0.05) inhibited expression of mRNA for IL-5. 5. The high topical potency of RPR 106541 in this model, which mimics important aspects of airway inflammation in human allergic asthmatics, suggests that this glucocorticoid may be useful in the treatment of bronchial asthma. PMID- 9351500 TI - Cyclo-oxygenase isozymes in mucosal ulcergenic and functional responses following barrier disruption in rat stomachs. AB - 1. We examined the effects of selective and nonselective cyclo-oxygenase (COX) inhibitors on various functional changes in the rat stomach induced by topical application of taurocholate (TC) and investigated the preferential role of COX isozymes in these responses. 2. Rat stomachs mounted in ex vivo chambers were perfused with 50 mM HCl and transmucosal potential difference (p.d.), mucosal blood flow (GMBF), luminal acid loss and luminal levels of prostaglandin E2 (PGE2) were measured before, during and after exposure to 20 mM TC. 3. Mucosal application of TC in control rats caused a reduction in p.d., followed by an increase of luminal acid loss and GMBF, and produced only minimal damage in the mucosa 2 h later. Pretreatment with indomethacin (10 mg kg[-1], s.c.), a nonselective COX-1 and COX-2 inhibitor, attenuated the gastric hyperaemic response caused by TC without affecting p.d. and acid loss, resulting in haemorrhagic lesions in the mucosa. In contrast, selective COX-2 inhibitors, such as NS-398 and nimesulide (10 mg kg[-1], s.c.), had no effect on any of the responses induced by TC and did not cause gross damage in the mucosa. 4. Luminal PGE2 levels were markedly increased during and after exposure to TC and this response was significantly inhibited by indomethacin but not by either NS-398 or nimesulide. The expression of COX-1-mRNA was consistently detected in the gastric mucosa before and after TC treatment, while a faint expression of COX-2-mRNA was detected only 2 h after TC treatment. 5. Both NS-398 and nimesulide significantly suppressed carrageenan-induced rat paw oedema, similar to indomethacin. 6. These results confirmed a mediator role for prostaglandins in the gastric hyperaemic response following TC-induced barrier disruption, and suggest that COX-1 but not COX-2 is a key enzyme in maintaining 'housekeeping' functions in the gastric mucosa under both normal and adverse conditions. PMID- 9351501 TI - Noradrenaline release from rat sympathetic neurones triggered by activation of B2 bradykinin receptors. AB - 1. The role of bradykinin receptors in the regulation of sympathetic transmitter release was investigated in primary cultures of neurones dissociated from superior cervical ganglia of neonatal rats. These cultures were loaded with [3H] noradrenaline and the outflow of radioactivity was determined under continuous superfusion. 2. Bradykinin (100 nmol l[-1] applied for 10 min) caused a transient increase in tritium outflow that reached a peak within four minutes after the beginning of the application and then declined towards the baseline, despite the continuing presence of the peptide. ATP (100 micromol l[-1]) and nicotine (10 micromol l[-1]) caused elevations in 3H outflow with similar kinetics, whereas outflow remained elevated during a 10 min period of electrical field stimulation (0.5 ms, 50 mA, 50 V cm[-1], 1.0 Hz). 3. When bradykinin was applied for periods of 2 min, the evoked 3H overflow was half-maximal at 12 nmol l(-1) and reached a maximum of 2.3% of cellular radioactivity. The preferential B1 receptor agonist des-Arg9-bradykinin failed to alter 3H outflow. The B2 receptor antagonists, [D Phe7]-bradykinin (1 micromol l[-1]) and Hoe 140 (10 nmol l[-1]), per se did not alter 3H outflow, but shifted the concentration-response curve for bradykinin evoked 3H overflow to the right by a factor of 7.9 and 4.3, respectively. 4. Bradykinin-induced overflow was abolished in the absence of extracellular Ca2+ and in the presence of either 1 micromol l(-1) tetrodotoxin or 300 micromol l(-1) Cd2+, as was electrically-induced overflow. Activation of alpha2-adrenoceptors by 1 micromol l(-1) UK 14,304 reduced both bradykinin- and electrically-triggered overflow. The Ca2+-ATPase inhibitor thapsigargin (0.3 micromol l[-1]) failed to alter either type of stimulated overflow. Caffeine (10 mmol l[-1]) enhanced bradykinin-induced overflow, but reduced overflow triggered by electrical field stimulation. 5. Inclusion of Ba2+ (0.1 to 1 mmol l[-1]) in the superfusion medium enhanced electrically induced overflow by approximately 100% and potentiated bradykinin-triggered overflow by almost 400%. Application of 1 mmol l(-1) Ba2+ for periods of 2 min triggered 3H overflow, and this overflow was abolished by 1 micromol l(-1) tetrodotoxin and enhanced by 10 mmol l(-1) caffeine. In contrast, inclusion of tetraethylammonium (0.1 to 1 mmol l[-1]) in the superfusion buffer caused similar increases of bradykinin- and electrically evoked 3H overflow (by about 100%), and tetraethylammonium, when applied for 2 min, failed to alter 3H outflow. 6. Treatment of cultures with 100 ng ml(-1) pertussis toxin caused a significant increase in bradykinin-, but not in electrically-, evoked tritium overflow. Treatment with 100 ng ml(-1) cholera toxin reduced both types of stimulated 3H overflow. 7. These data reveal bradykinin as a potent stimulant of action potential-mediated and Ca2+-dependent transmitter release from rat sympathetic neurones in primary cell culture. This neurosecretory effect of bradykinin involves activation of B2-receptors, presumably linked to pertussis- and cholera toxin-insensitive G proteins, most likely members of the Gq family. Results obtained with inhibitors of muscarinic K+ (KM) channels, like caffeine and Ba2+, indicate that the secretagogue action of bradykinin probably involves inhibition of these K+ channels. PMID- 9351502 TI - Inhibitory effects of (+/-)-propranolol on excitation-contraction coupling in isolated soleus muscles of the rat. AB - 1. The effect of a beta-adrenoceptor antagonist, propranolol, was investigated on excitation-contraction coupling in small, intact bundles of soleus muscle fibres from the rat. 2. (+/-)-Propranolol significantly inhibited twitch and tetanic tension with IC50 values of 6.7 microM and 3.5 microM, respectively. 3. (+) Propranolol (which has 100 times less beta-blocking activity than the (+/-) form) was approximately one third as effective as the (+/-) form at inhibiting isometric tension. 4. (+/-)-Propranolol (20 microM) had no significant effect on the amplitude of caffeine contractures, suggesting that it did not directly inhibit Ca2+ release from the sarcoplasmic reticulum. 5. The resting membrane potential measured after 15 min perfusion with 20 microM (+/-)-propranolol was not significantly different from control. However, this concentration of (+/-) propranolol significantly reduced both the peak amplitude and the maximum rate of rise of the action potential. Both effects were only partially reversible after extensive washing. 6. (+/-)-Propranolol perfusion caused a modest reduction in the amplitude of sub-maximal K+ contractures at concentrations (5 microM) that markedly depressed tetanic tension. 7. The results indicate that (+/-) propranolol can decrease isometric tension independently of beta-receptor occupation by (i) reducing the amplitude and rate of rise of the action potential and (ii) by directly inhibiting excitation-contraction coupling. The relatively low IC50 for the 'membrane-stabilizing' action of propranolol on tetanic tension (3.5 microM), combined with the ability of the drug to accumulate gradually in biological membranes, may contribute to a peripheral component of the tremorolytic and fatigue-inducing actions of propranolol on skeletal muscle. PMID- 9351503 TI - In vitro and in vivo characterization of NK3 receptors in the rabbit eye by use of selective non-peptide NK3 receptor antagonists. AB - 1. Inhibition of NK3 receptor agonist-induced contraction in the rabbit isolated iris sphincter muscle was used to assess the in vitro functional activity of three 2-phenyl-4-quinolinecarboxamides, members of a novel class of potent and selective non-peptide NK3 receptor antagonists. In addition, an in vivo correlate of this in vitro response, namely NK3 receptor agonist-induced miosis in conscious rabbits, was characterized with some of these antagonists. 2. In vitro senktide (succinyl-[Asp9,MePhe8]-substance P (6-11) and [MePhe7]-neurokinin B ([MePhe7]-NKB) were potent contractile agents in the rabbit iris sphincter muscle but exhibited quite different profiles. Senktide produced monophasic log concentration-effect curves with a mean pD2=9.03+/-0.06 and mean nH=1.2+/-0.02 (n=14). In contrast, [MePhe7]-NKB produced shallow log concentration-effect curves which often appeared biphasic (nH=0.54+/-0.04, n=8), preventing the accurate determination of pD2 values. 3. The contractile responses to the NK3 receptor agonist senktide were antagonized in a surmountable and concentration dependent manner by SB 223412 ((-)-(S)-N-(alpha-ethylbenzyl)-3-hydroxy-2 phenylquinoline-4-ca rboxamide; 3-30 nM, pA2=8.4, slope=1.8+/-0.3, n=4). SB 222200 ((-)-(S)-N-(alpha-ethylbenzyl)-3-methyl-2-phenylquinoline-4-car box amide; 30-300 nM, pA2=7.9, slope=1.4+/-0.06, n=4) and SB 218795 ((-)-(R)-N-(alpha methoxycarbonylbenzyl)-2-phenylquinoline-4-carboxamide; 0.3 and 3 microM apparent pKB=7.4+/-0.06, n=6). 4. Contractile responses to the NK3 receptor agonist [MePhe7]-NKB in the rabbit iris sphincter muscle were unaffected by SB 218795 (0.3 and 3 microM, n=8). In contrast, SB 223412 (30 and 300 microM n=4) and SB 222200 (0.3 and 3 microM, n=4) inhibited responses to low concentrations (< or = 1 nM), to a greater extent than higher concentrations (> 1 nM) of [MePhe7]-NKB. Furthermore, log concentration-effect curves to [MePhe7]-NKB became steeper and monophasic in the presence of each antagonist. 5. SB 218795 (3 microM, n=4) had no effect on contractions induced by transmural nerve stimulation (2 Hz) or substance P, exemplifying the selectivity of this class of antagonist for functional NK3 receptors over NK1 receptors in the rabbit. 6. In vivo, senktide (1, 10 and 25 microg i.v., i.e. 1.2, 11.9 and 29.7 nmol, respectively) induced concentration-dependent bilateral miosis in conscious rabbits (maximum pupillary constriction=4.25+/-0.25 mm; basal pupillary diameter 7.75+/-0.48 mm; n=4). The onset of miosis was within 2-5 min of application of senktide and responses lasted up to 30 min. Responses to two i.v. administrations of 25 microg senktide given 30 min apart revealed no evidence of tachyphylaxis. Topical administration of atropine (1%) to the eye enhanced pupillary responses to 25 microg senktide. This was probably due to the mydriatic effect of atropine since it significantly increased baseline pupillary diameter from 7.0+/-0.4 mm to 9.0+/-0.7 mm (n=4), thereby increasing the maximum capacity for miosis. Senktide-induced miosis was inhibited by SB 222200 (1 and 2 mg kg[-1], i.v., i.e. 2.63 and 5.26 micromol kg[ 1]; maximum inhibition 100%; n=3-4), SB 223412 (0.5 and 1 mg kg[-1], i.v., i.e. 1.31 and 2.61 micromol kg[-1]; maximum inhibition 100%; n=3), SB 218795 (0.5 and 1 mg kg[-1] i.v., i.e. 1.26 and 2.52 micromol kg-1; maximum inhibition 78%; n=3), and the structurally distinct NK3 receptor antagonist SR 142801 ((S)-(N)-(1-(3-(1 benzoyl-3-(3,4-dichlorophenyl)piperidin-3-yl)propyl)-4-phenylepipiperidin-4-yl)-N methylacetamide; 1.5mg kg-1, i.v., i.e. 2.47micromol kg-1, maximum inhibition 92%; n=3). 7. Topical administration of senktide (25microg; 29.7nmol) to the eye induced unilateral miosis in the treated eye only. At this dose there was no significant difference (P<0.05) between pupillary constriction obtained by topical or i.v. senktide, and topically administered atropine had no significant effect on responses to topical senktide (n=4). 8. [MePhe7]-NKB (125, 250 and 500microg, i.v., i.e. 98.31, 196.62 and 393.24nmol, respectively) also induced bilateral miosis in conscious rabbits (maximum pupillary constriction=4.13+/ 0.30mm; n=4), but in contrast to in vitro studies this agonist was approximately 100 fold less potent than senktide. [MePhe7]-NKB-induced miosis was inhibited by SB 222200 (5mg kg-1, i.v., i.e. 13.14micromol kg-1; maximum inhibition 69%; n=3). 9. In summary, SB 223412, SB 222200 and SB 218795 are potent and selective antagonists of NK3 receptor-mediated contraction in the rabbit isolated iris sphincter muscle. In addition, NK3 receptor agonist-induced miosis in conscious rabbits is a good in vivo correlate of the in vitro rabbit iris sphincter muscle preparation and appears to be a useful model for characterizing the pharmacodynamic profile and efficacy of structurally distinct NK3 receptor antagonists, such as SB 222200, SB 223412, SB 218795 and SR 142801. PMID- 9351504 TI - Characterization of action potential-triggered [Ca2+]i transients in single smooth muscle cells of guinea-pig ileum. AB - 1. To characterize increases in cytosolic free Ca2+ concentration ([Ca2+]i) associated with discharge of action potentials, membrane potential and [Ca2+]i were simultaneously recorded from single smooth muscle cells of guinea-pig ileum by use of a combination of nystatin-perforated patch clamp and fura-2 fluorimetry techniques. 2. A single action potential in response to a depolarizing current pulse elicited a transient rise in [Ca2+]i. When the duration of the current pulse was prolonged, action potentials were repeatedly discharged during the early period of the pulse duration with a progressive decrease in overshoot potential, upstroke rate and repolarization rate. However, such action potentials could each trigger [Ca2+]i transients with an almost constant amplitude. 3. Nicardipine (1 microM) and La3+ (10 microM), blockers of voltage-dependent Ca2+ channels (VDCCs), abolished both the action potential discharge and the [Ca2+]i transient. 4. Charybdotoxin (ChTX, 300 nM) and tetraethylammonium (TEA, 2 mM), blockers of large conductance Ca2+-activated K+ channels, decreased the rate of repolarization of action potentials but increased the amplitude of [Ca2+]i transients. 5. Thapsigargin (1 microM), an inhibitor of SR Ca2+-ATPase, slowed the falling phase and somewhat increased the amplitude, of action potential triggered [Ca2+]i transients without affecting action potentials. In addition. in voltage-clamped cells, the drug had little effect on the voltage step-evoked Ca2+ current but exerted a similar effect on its concomitant rise in [Ca2+]i to that on the action potential-triggered [Ca2+]i transient. 6. Similar action potential triggered [Ca2+]i transients were induced by brief exposures to high-K+ solution. They were not decreased, but rather increased, after depletion of intracellular Ca2+ stores by a combination of ryanodine (30 microM) and caffeine (10 mM) through an open-lock of Ca2+-induced Ca2+ release (CICR)-related channels. 7. The results show that action potentials, discharged repeatedly during the early period of a long membrane depolarization, undergo a progressive change in configuration but can each trigger a constant rise in [Ca2+]i. Intracellular Ca2+ stores have a role, especially in accelerating the falling phase of the action potential-triggered [Ca2+]i transients by replenishing cytosolic Ca2+. No evidence was provided for the involvement of CICR in the action potential triggered [Ca2+]i transient. PMID- 9351505 TI - Effects of ibuprofen enantiomers and its coenzyme A thioesters on human prostaglandin endoperoxide synthases. AB - 1. Ibuprofen enantiomers and their respective coenzyme A thioesters were tested in human platelets and blood monocytes to determine their selectivity and potency as inhibitors of cyclo-oxygenase activity of prostaglandin endoperoxide synthase 1 (PGHS-1) and PGHS-2. 2. Human blood from volunteers was drawn and allowed to clot at 37 degrees C for 1 h in the presence of increasing concentrations of the test compounds (R-ibuprofen, S-ibuprofen, R-ibuprofenoyl-CoA, S-ibuprofenoyl-CoA, NS-398). Immunoreactive (ir) thromboxane B2 (TXB2) concentrations in serum were determined by a specific EIA assay as an index of the cyclo-oxygenase activity of platelet PGHS-1. 3. Heparin-treated blood from the same donors was incubated at 37 degrees C for 24 h with the same concentrations of the test compounds in the presence of lipopolysaccharide (LPS, 10 microg ml[-1]). The contribution of PGHS 1 was suppressed by pretreatment of the volunteers with aspirin (500 mg; 48 h before venepuncture). As a measure of LPS induced PGHS-2 activity immunoreactive prostaglandin E2 (irPGE2) plasma concentrations were determined by a specific EIA assay. 4. S-ibuprofen inhibited the activity of PGHS-1 (IC50 2.1 microM) and PGHS 2 (IC50 1.6 microM) equally. R-ibuprofen inhibited PGHS-1 (IC50 34.9) less potently than S-ibuprofen and showed no inhibition of PGHS-2 up to 250 microM. By contrast R-ibuprofenoyl-CoA thioester inhibited PGE2 production from LPS stimulated monocytes almost two orders of magnitude more potently than the generation of TXB2 (IC50 5.6 vs 219 microM). 5. Western blotting of PGHS-2 after LPS induction of blood monocytes showed a concentration-dependent inhibition of PGHS-2 protein expression by ibuprofenoyl-CoA thioesters. 6. These data confirm that S-ibuprofen represents the active entity in the racemate with respect to cyclo-oxygenase activity. More importantly the data suggest a contribution of the R-enantiomer to therapeutic effects not only by chiral inversion to S-ibuprofen but also via inhibition of induction of PGHS-2 mediated by R-ibuprofenoyl-CoA thioester. 7. The data may explain why racemic ibuprofen is ranked as one of the safest non-steroidal anti-inflammatory drugs (NSAIDs) so far determined in epidemiological studies. PMID- 9351506 TI - Role of peroxynitrite and activation of poly (ADP-ribose) synthase in the vascular failure induced by zymosan-activated plasma. AB - 1. Zymosan is a wall component of the yeast Saccharomyces Cerevisiae. Injection of zymosan into experimental animals is known to produce an intense inflammatory response. Recent studies demonstrated that the zymosan-induced inflammatory response in vivo can be ameliorated by inhibitors of nitric oxide (NO) biosynthesis. The cytotoxic effects of NO are, in part, mediated by the oxidant preoxynitrite and subsequent activation of the nuclear enzyme poly (ADP-ribose) synthetase (PARS). In the present in vitro study, we have investigated the cellular mechanisms of vascular failure elicited by zymosan-activated plasma and the contribution of peroxynitrite production and activation of PARS to the changes. 2. Incubation of rat aortic smooth muscle cells with zymosan-activated plasma (ZAP) induced the production of nitrite, the breakdown product of NO, due to the expression of the inducible isoform of NO synthase (iNOS) over 6 24 h. In addition, ZAP triggered the production of peroxynitrite in these cells, as measured by the oxidation of the fluorescent dye dihydrorhodamine 123 and by nitrotyrosine Western blotting. 3. Incubation of the smooth muscle cells with ZAP induced DNA single strand breakage and PARS activation. These effects were reduced by inhibition of NOS with NG-methyl-L-arginine (L-NMA, 3 mM), and by glutathione (3 mM), a scavenger of peroxynitrite. The PARS inhibitor 3 aminobenzamide (1 mM) inhibited the ZAP-induced activation of PARS. 4. Incubation of thoracic aortae with ZAP in vitro caused a reduction of the contractions of the blood vessels to noradrenaline (vascular hyporeactivity) and elicited a reduced responsiveness to the endothelium-dependent vasodilator acetylcholine (endothelial dysfunction). 5. Preincubation of the thoracic aortae with L-NMA (1 mM), glutathione (3 mM) or by the PARS inhibitor 3-aminobenzamide (1 mM) prevented the development of vascular hyporeactivity in response to ZAP. Moreover, glutathione and 3-aminobenzamide treatment protected against the ZAP induced development of endothelial dysfunction. The PARS-related loss of the vascular contractility was evident at 30 min after incubation in endothelium intact, but not in endothelium-denuded vessels and also manifested at 6 h after incubation with ZAP in endothelium-denuded rings. The acute response is probably related, therefore, to peroxynitrite formation (involving the endothelial NO synthase), whereas the delayed response may be related to the expression of iNOS in the smooth muscle. 6. The data obtained suggest that zymosan-activated plasma causes vascular dysfunction by inducing the simultaneous formation of superoxide and NO. These radicals combine to form peroxynitrite, which, in turn causes DNA injury and PARS activation. The protective effect of 3-aminobenzamide demonstrates that PARS activation contributes both to the development of vascular hyporeactivity and endothelial dysfunction during the vascular failure induced by ZAP. PMID- 9351507 TI - Actions of 4-chloro-3-ethyl phenol on internal Ca2+ stores in vascular smooth muscle and endothelial cells. AB - 1. Recently, 4-chloro-3-ethyl phenol (CEP) has been shown to cause the release of internally stored Ca2+ apparently through ryanodine-sensitive Ca2+ channels, in fractionated skeletal muscle terminal cisternae and in a variety of non-excitable cell types. Its action on smooth muscle is unknown. In this study, we characterized the actions of CEP on vascular contraction in endothelium-denuded dog mesenteric artery. We also determined its ability to release Ca2+, by use of Ca2+ imaging techniques, on dog isolated mesenteric artery smooth muscle cells and on bovine cultured pulmonary artery endothelial cells. 2. After phenylephrine (PE, 10 microM) sensitive Ca2+ stores were depleted by maximal PE stimulation in Ca2+-free medium, the action of CEP on refilling of the emptied PE stores was tested, by first pre-incubating the endothelium-denuded artery in CEP for 15 min before Ca2+ was restored for a 30 min refilling period. At the end of this period, Ca2+ and CEP were removed, and the arterial ring was tested again with PE to assess the degree of refilling of the internal Ca2+ store. 3. In a concentration-dependent manner (30, 100 and 300 microM), CEP significantly reduced the size of the post-refilling PE contraction (49.4, 28.9 and 5.7% of control, respectively) in Ca2+-free media. This suggests that Ca2+ levels are reduced in the internal stores by CEP treatment. CEP alone did not cause any contraction either in Ca2+-containing or Ca2+-free Krebs solution. 4. Restoring Ca2+ in the presence of PE caused a large contraction, which reflects PE-induced influx of extracellular Ca2+. The contraction of tissues pretreated with 300 microM CEP was significantly less compared with controls. However, tissues pretreated with 30 and 100 microM CEP were unaffected. Washout of CEP over 30 min produced complete recovery of responses to PE in Ca2+-free and Ca2+-containing medium suggesting a rapid reversal of CEP effects. 5. Concentration-response curves were constructed for PE, 5-hydroxytryptamine (5-HT) and K+ in the absence of and after 30 min pre-incubation with 30, 100 and 300 microM CEP. In all cases, CEP caused a concentration-dependent depression of the maximum response to PE (84.8, 43.4 and 11.6% of control), 5-HT (65.4, 25.7 and 6.9% of control) and K+ (77.6, 41.1 and 10.8% of control). 6. Some arterial rings were pre-incubated with ryanodine (30 microM) for 30 min before the construction of PE concentration response curves. In Ca2+-free Krebs solution, ryanodine alone did not cause any contraction. However, 58% (11 out of 19) of the tissues tested with ryanodine developed contraction (6.9+/-1.2% of 100 mM K+ contraction, n=11) in the presence of external Ca2+. EC50 values for PE in ryanodine-treated tissues (1.7+/-0.25 microM, n=16) were not significantly different from controls (2.5+/-0.41 microM, n=22). Maximum contractions to PE (118.5+/-4.4% of 100 mM K+ contraction, n=16) were also unaffected by ryanodine when compared to controls (129+/-4.2%, n=23). 7. When fura-2 loaded smooth muscle cells (n=13) and endothelial cells (n=27) were imaged for Ca2+ distribution, it was observed that 100 and 300 microM CEP in Ca2+-free medium caused Ca2+ release in both cell types. Smooth muscle cells showed a small decrease in cell length. Addition of EGTA (5 mM) reversed the effect of CEP on intracellular Ca2+ to control values. 8. These data show, for the first time in vascular smooth muscle and endothelial cells, that CEP releases Ca2+ more rapidly than ryanodine. Unlike ryanodine, CEP caused no basal contraction but depressed contractions to PE, 5-HT and K+. The lack of basal contraction may result from altered responsiveness of the contractile system to intracellular Ca2+ elevation. PMID- 9351508 TI - Presynaptic inhibition of synaptic transmission in the rat hippocampus by activation of muscarinic receptors: involvement of presynaptic calcium influx. AB - 1. Modulation of presynaptic voltage-dependent calcium channels (VDCCs) by muscarinic receptors at the CA3-CA1 synapse of rat hippocampal slices was investigated by using the calcium indicator fura-2. Stimulation-evoked presynaptic calcium transients ([Ca(pre)]t) and field excitatory postsynaptic potentials (fe.p.s.ps) were simultaneously recorded. The relationship between presynaptic calcium influx and synaptic transmission was studied. 2. Activation of muscarinic receptors inhibited [Ca(pre)]t, thereby reducing synaptic transmission. Carbachol (CCh, 10 microM) inhibited [Ca(pre)]t by 35% and reduced fe.p.s.p. by 85%. The inhibition was completely antagonized by 1 microM atropine. An approximate 4th power relationship was found between presynaptic calcium influx and postsynaptic responses. 3. Application of the N-type VDCC-blocking peptide toxin omega-conotoxin GVIA (omega-CTx GVIA, 1 microM) inhibited [Ca(pre)]t and fe.p.s.ps by 21% and 49%, respectively, while the P/Q-type VDCC blocker omega-agatoxin IVA (omega)-Aga IVA, 1 microM) reduced [Ca(pre)]t and fe.p.s.ps by 35% and 85%, respectively. 4. Muscarinic receptor activation differentially inhibited distinct presynaptic VDCCs. Omega-CTx GVIA-sensitive calcium channels were inhibited by muscarinic receptors, while omega-Aga IVA sensitive channels were not. The percentage inhibition of omega-CTx GVIA sensitive [Ca(pre)]t was about 63%. 5. Muscarinic receptors inhibited presynaptic VDCCs in a way similar to adenosine (Ad) receptors. The percentage inhibition of omega-CTx GVIA-sensitive [Ca(pre)]t by Ad (100 microM) was about 59%. There was no significant inhibition of omega-Aga IVA-sensitive channels by Ad. The inhibitions of [Ca(pre)]t by CCh and Ad were mutually occlusive. 6. These results indicate that inhibition of synaptic transmission by muscarinic receptors is mainly the consequence of a reduction of the [Ca(pre)]t due to inhibition of presynaptic VDCCs. PMID- 9351509 TI - Kappa1- and kappa2-opioid receptors mediating presynaptic inhibition of dopamine and acetylcholine release in rat neostriatum. AB - 1. The effects of selective opioid receptor agonists and antagonists on N-methyl D-aspartate (NMDA, 10 microM)-induced release of [3H]-dopamine and [14C] acetylcholine (ACh) from superfused neostriatal slices were studied to investigate the possible occurrence of functional kappa-opioid receptor subtypes in rat brain. 2. The kappa receptor agonists (-)-ethylketocyclazocine ((-)-EKC), U69593 and the endogenous opioid peptide dynorphin A1-13 caused a naloxone reversible inhibition of NMDA-induced [3H]-dopamine release, with pD2 values of about 9, 8.5 and 8.2, respectively, whereas both the mu agonist Tyr-D-Ala-Gly (NMe)Phe-Gly-ol (DAMGO) and the delta agonist D-Pen2-D-Pen5-enkephalin (DPDPE) were ineffective in this respect. The inhibitory effect of submaximally effective concentrations of dynorphin A1-13, U69593 and (-)-EKC on NMDA-induced [3H] dopamine release were not changed by the delta1/delta2-opioid receptor antagonist naltrindole (up to a concentration of 1 microM, but reversed by the kappa receptor antagonist nor-binaltorphimine (nor-BNI), with an IC50) as low as 0.02 nM, indicating the involvement of U69593-sensitive kappa1-opioid receptors. 3. NMDA-induced [14C]-ACh release was reduced in a naloxone-reversible manner by DPDPE (pD2 about 7.2), dynorphin A1-13 (pD2 6.7) and EKC (pD2 6.2), but not by U69593 and DAMGO. The inhibitory effect of a submaximally effective concentration of DPDPE, unlike those of dynorphin A1-13 and (-)-EKC, on NMDA-induced [14C]-ACh release was antagonized by naltrindole with an IC50 of 1 nM, indicating the involvement of delta-opioid receptors in the inhibitory effect of DPDPE. On the other hand, the inhibitory effects of dynorphin A1-13 and (-)-EKC on [14C]-ACh release were readily antagonized by nor-BNI with an IC50 of about 3 nM. A 100 fold higher concentration of nor-BNI also antagonized the inhibitory effect of DPDPE, indicating the involvement of U69593-insensitive kappa2-opioid receptors in the inhibitory effects of dynorphin A1-13 and (-)-EKC. 4. Although naloxone benzoylhydrazone (NalBzoH), displaying high affinity towards the putative kappa3 opioid receptor, antagonized the inhibitory effects of dynorphin A1-13 and (-) EKC on [3H]-dopamine and [14C]-ACh release as well as that of U69593 on [3H] dopamine release, it displayed a low apparent affinity (IC50 about 100 nM) in each case. 5. In conclusion, whereas activation of kappa1-opioid receptors causes presynaptic inhibition of NMDA-induced dopamine release, kappa2 receptor activation results in inhibition of ACh release in rat neostriatum. As such, this study is the first to provide unequivocal in vitro evidence for the existence of functionally distinct kappa-opioid receptor subtypes in the brain. PMID- 9351510 TI - Pharmacokinetic-pharmacodynamic modelling of the anti-lipolytic and anti-ketotic effects of the adenosine A1-receptor agonist N6-(p-sulphophenyl)adenosine in rats. AB - 1. The purpose of this study was to develop and validate an integrated pharmacokinetic-pharmacodynamic model for the anti-lipolytic effects of the adenosine A1-receptor agonist N6-(p-sulphophenyl)adenosine (SPA). Tissue selectivity of SPA was investigated by quantification of haemodynamic and anti lipolytic effects in individual animals. 2. After intravenous infusion of SPA to conscious normotensive Wistar rats, arterial blood samples were drawn for determination of blood SPA concentrations, plasma non-esterified fatty acid (NEFA) and beta-hydroxybutyrate levels. Blood pressure and heart rate were monitored continuously. 3. The relationship between the SPA concentrations and the NEFA lowering effect was described by the indirect suppression model. Administration of SPA at different rates and doses (60 microg kg[-1] in 5 min and 15 min, and 120 microg kg[-1] in 60 min) led to uniform pharmacodynamic parameter estimates. The averaged parameters (mean+/-s.e., n=19) were Emax: -80+/-2% (% change from baseline), EC50: 22+/-2 ng ml(-1), and Hill factor: 2.2+/-0.2. 4. In another group, given 400 microg kg(-1) SPA in 15 min, pharmacodynamic parameters for both heart rate and anti-lipolytic effect were derived within the same animal. The reduction in heart rate was directly related to blood concentration on the basis of the sigmoidal Emax model. SPA inhibited lipolysis at concentrations lower than those required for an effect on heart rate. The EC50 values (mean+/-s.e., n=6) were 131+/-31 ng ml(-1) and 20+/-3 ng ml(-1) for heart rate and NEFA lowering effect, respectively. 5. In conclusion, the relationship between blood SPA concentrations and anti-lipolytic effect was adequately described by the indirect suppression model. For SPA a 6 fold difference in potency was observed between the effects on heart rate and NEFAs, indicating some degree of tissue selectivity in vivo. PMID- 9351511 TI - Differential effect of dexamethasone on interleukin 1beta- and cyclic AMP triggered expression of GTP cyclohydrolase I in rat renal mesangial cells. AB - 1. Endogenous synthesis of tetrahydrobiopterin (BH4) is an essential requirement for cytokine-stimulated nitric oxide (NO) synthesis in rat mesangial cells. GTP cyclohydrolase I, the rate-limiting enzyme in BH4 synthesis, is expressed in renal mesangial cells in response to two principal classes of activating signals. These two groups of activators comprise inflammatory cytokines such as interleukin (IL)-1beta and agents that elevate cellular levels of cyclic AMP. 2. We examined the action of the potent anti-inflammatory drug dexamethasone on GTP cyclohydrolase I induction in response to IL-1beta and a membrane-permeable cyclic AMP analogue, N6, O-2'-dibutyryladenosine 3'-5'-phosphate (Bt2cyclic AMP). 3. Nanomolar concentrations of dexamethasone markedly attenuated IL-1beta-induced GTP cyclohydrolase I mRNA steady state level as well as IL-1beta-induced GTP cyclohydrolase I protein expression and enzyme activity. In contrast, dexamethasone did not inhibit Bt2cyclic AMP-triggered increase in GTP cyclohydrolase I mRNA level and protein expression, and low (1 nM) or high (1 and 10 microM) doses of dexamethasone consistently increased Bt2cyclic AMP-induced GTP cyclohydrolase activity. 4. In summary, these results suggest that glucocorticoids act at several levels, critically dependent on the stimulus used, to control GTP cyclohydrolase I expression. PMID- 9351512 TI - Effect of chronic treatment with the GABA transaminase inhibitors gamma-vinyl GABA and ethanolamine O-sulphate on the in vitro GABA release from rat hippocampus. AB - 1. The effects of 2, 8 and 21 day oral treatment with the specific gamma aminobutyric acid transaminase (GABA-T) inhibitors gamma-vinyl GABA (GVG) and ethanolamine O-sulphate (EOS) on brain GABA levels, GABA-T activity, and basal and stimulated GABA release from rat cross-chopped brain hippocampal slices was investigated. 2. Treatment with GABA-T inhibitors lead to a reduction in brain GABA-T activity by 65-80% compared with control values, with a concomitant increase in brain GABA content of 40-100%. 3. Basal hippocampal GABA release was increased to 250-450% of control levels following inhibition of GABA-T activity. No Ca2+ dependence was observed in either control or treated tissues. 4. GVG and EOS administration led to a significant elevation in the potassium stimulated release of GABA from cross-chopped hippocampal slices compared with that of controls. Although stimulated GABA release from control tissues was decreased in the presence of a low Ca2+ medium, GVG and EOS treatment abolished this Ca2+ dependency. 5. GABA compartmentalization, Na+ and Cl- coupled GABA uptake carriers and glial release may provide explanations for the loss of the Ca2+ dependency of stimulated GABA release observed following GVG and EOS treatment. 6. Administration of GABA-T inhibitors led to increases in both basal and stimulated hippocampal GABA release. However, it is not clear which is the most important factor in the anticonvulsant activity of these drugs, the increased GABA content 'leaking' out of neurones and glia leading to widespread inhibition, or the increase in stimulated GABA release which may occur following depolarization caused by an epileptic discharge. PMID- 9351513 TI - An endogenous A2B adenosine receptor coupled to cyclic AMP generation in human embryonic kidney (HEK 293) cells. AB - 1. Cyclic AMP generation by adenosine analogues was examined in human embryonic kidney (HEK 293) cells by use of a [3H]-adenine pre-labelling methodology. 2. Adenosine analogues showed the following rank order of potency (pD2 value): 5'-N ethylcarboxamidoadenosine (NECA, 5.24)>2-chloroadenosine (4.41) > or = adenosine (4.19)= N6-(2-(4-aminophenyl)-ethylamino)adenosine (APNEA, 4.11). The A2A selective agonist CGS21680 failed to elicit a significant stimulation of cyclic AMP generation at concentrations below 30 microM. 3. Of these agents, NECA was observed to exhibit the greatest intrinsic activity, while in comparison maximal responses to adenosine (76+/-8% NECA response), 2-chloroadenosine (70+/-6%) and APNEA (40+/-3%) were significantly reduced. 4. Antagonists of the NECA-evoked cyclic AMP generation showed the rank order of apparent affinity (apparent pA2 value): CGS 15943 (7.79)=XAC (7.74)>DPCPX (7.01)=PD115199 (6.93) 8FB-PTP (6.80)>KF 17837 (5.98)>3-propylxanthine (5.13). 5. Agarose gel electrophoresis of the products of the polymerase chain reaction, with cDNA generated from HEK 293 cell total RNA showed virtually identical patterns and nucleotide sizes in comparison with the vector for the full length human brain A2B adenosine receptor. 6. We concluded that HEK 293 cells express an endogenous adenosine receptor coupled to cyclic AMP generation which is of the A2B subtype. PMID- 9351514 TI - Anti-inflammatory activity of cationic lipids. AB - 1. The effect of liposome phospholipid composition has been assumed to be relatively unimportant because of the presumed inert nature of phospholipids. 2. We have previously shown that cationic liposome formulations used for gene therapy inhibit, through their cationic component, the synthesis by activated macrophages of the pro-inflammatory mediators nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha). 3. In this study, we have evaluated the ability of different cationic lipids to reduce footpad inflammation induced by carrageenan and by sheep red blood cell challenge. 4. Parenteral (i.p. or s.c) or local injection of the positively charged lipids dimethyldioctadecylammomium bromide (DDAB), dioleyoltrimethylammonium propane (DOTAP), dimyristoyltrimethylammonium propane (DMTAP) or dimethylaminoethanecarbamoyl cholesterol (DC-Chol) significantly reduced the inflammation observed in both models in a dose-dependent manner (maximum inhibition: 70-95%). 5. Cationic lipids associated with dioleyol- or dipalmitoyl-phosphatidylethanolamine retained their anti-inflammatory activity while cationic lipids associated with dipalmitoylphosphatidylcholine (DPPC) or dimyristoylphosphatidylglycerol (DMPG) showed no anti-inflammatory activity, indicating that the release of cationic lipids into the macrophage cytoplasm is a necessary step for anti-inflammatory activity. The anti-inflammatory activity of cationic lipids was abrogated by the addition of dipalmitoylphosphatidylethanolamine-poly(ethylene)glycol-2000 (DPPE PEG2000) which blocks the interaction of cationic lipids with macrophages. 6. Because of the significant role of protein kinase C (PKC) in the inflammatory process we have determined whether the cationic lipids used in this study inhibit PKC activity. The cationic lipids significantly inhibited the activity of PKC but not the activity of a non-related protein kinase, PKA. The synthesis of interleukin-6 (IL-6), which is not dependent on PKC activity for its induction in macrophages, was not modified in vitro or in situ by cationic lipids. The synthesis of NO and TNF-alpha in macrophages, both of which are PKC-dependent, was downregulated by cationic lipids. 7. These results demonstrate that cationic lipids can be considered as novel anti-inflammatory agents. The downregulation of pro-inflammatory mediators through interaction of cationic lipids with the PKC pathway may explain this anti-inflammatory activity. Furthermore, since cationic lipids have intrinsic anti-inflammatory activity, cationic liposomes should be used with caution to deliver nucleic acids for gene therapy in vivo. PMID- 9351515 TI - The signalling pathway which causes contraction via P2-purinoceptors in rat urinary bladder smooth muscle. AB - 1. The signalling pathway which causes contractions to adenosine 5'-O-2 thiodiphosphate (ADPbetaS) and alpha,beta-methylene adenosine 5'-diphosphate (alpha,beta-Me ADP) was investigated in rat urinary bladder smooth muscle by measuring isotonic tension. 2. The responses to 10 microM alpha,beta-methylene adenosine 5'-triphosphate (alpha,beta-Me ATP) in 0 and 3.6 mM Ca2+ were 5.9+/-1.3 (n=10) and 122.2+/-6.4 (n=8) % respectively of those obtained in 1.8 mM Ca2+, whereas those to 100 microM ADPbetaS were 34.6+/-3.3 (n=8) and 96.8+/-7.2 (n = 8) %, in 0 and 3.6 mM Ca2+, respectively. In both experimental conditions, the responses to the two agonists expressed as % of the control responses were significantly different (P<0.01). 3. Indomethacin at high concentrations (>1 microM) decreased the responses to alpha,beta-Me ATP (10 microM), ADPbetaS (100 microM) and alpha,beta-Me ADP (100 microM). However, no significant difference was obtained between the responses to all the agonists at 30 microM indomethacin. 4. 2-Nitro-4-carboxphenyl n,n-diphenylcarbamate (NCDC) at concentrations between 1 microM and 100 microM concentration-dependently decreased the responses to ADPbetaS (100 microM) and alpha,beta-Me ADP (100 microM) and almost completely inhibited them at 100 microM. Although the responses to alpha,beta-Me ATP (10 microM) were also inhibited by the drug, at 50 and 100 microM NCDC the responses to alpha,beta-Me ATP were significantly larger than those to ADPbetaS and alpha,beta-Me ADP (P<0.01). 5.NCDC 100 microM significantly inhibited the KCl induced contraction to 65.9+/-4.9% (n=6) of the control (P<0.01). 6. It is suggested that the contraction via ADPbetaS-sensitive receptors in the rat urinary bladder smooth muscle mainly depends on Ca2+ ions liberated from intracellular Ca2+ stores, though the contribution of Ca2+ ions from the extracellular space cannot be neglected. The release of Ca2+ ions from stores is mainly mediated by the production of inositol trisphosphate (IP3) via the activation of phospholipase C. PMID- 9351516 TI - Comparison of the acute cardiotoxicity of the antimalarial drug halofantrine in vitro and in vivo in anaesthetized guinea-pigs. AB - 1. Several unrelated drugs have pro-arrhythmic activity associated with an ability to prolong the QT interval of the ECG. The aim of this work was to examine the effects of the antimalarial drug halofantrine in vivo and in vitro. 2. In anaesthetized guinea-pigs consecutive bolus doses of halofantrine (0.3, 1, 3, 10 and 30 mg kg(-1), i.v.) at 25 min intervals caused dose-dependent prolongation of the rate corrected QTc interval and bradycardia. The change in heart rate became significant after administration of 10 mg kg(-1) halofantrine ( 23+/-9 beats min[-1]) whereas the increase in QTc was significant with only 1 mg kg(-1) halofantrine (22+/-10 ms). It was only with the highest dose of halofantrine that the PR interval was increased (from 52+/-3 to 67+/-4 ms) and second degree atrioventricular (AV) block (type 1 Mobitz) occurred in all animals. No changes were observed in any parameters in a separate group of guinea pigs which received vehicle (dimethylacetamide 60% propylene glycol 40%) at equivalent time points. 3. The blood concentrations of halofantrine ranged from 0.26+/-0.17 microM after administration of 0.3 mg kg(-1) to 2.79+/-0.87 microM after 30 mg kg(-1), i.v. There was a significant correlation between the blood concentrations of halofantrine and the changes in QTc interval. 4 In guinea-pig left papillary muscles the effective refractory period was increased significantly 60 min after addition of halofantrine; from 161+/-4 to 173+/-6 ms with 10 microM, 156+/-8 to 174+/-6 ms with 30 microM and 165+/-6 to 179+/-5 ms with 100 microM halofantrine. However, the vehicle (0.1% Tween 80 in DMSO; final concentration of vehicle in Krebs, 1%) also increased the effective refractory period from 164+/-5 to 173+/-6 ms. Similar results were obtained in right ventricular strips but left atrial effective refractory periods were not altered by either the vehicle or halofantrine. 5. The results of these experiments suggest that any direct effects that halofantrine may have had on the effective refractory period of cardiac muscle cannot be separated from those of the vehicle. The prolongation of QTc and consistent observation of AV block with halofantrine in anaesthetized guinea-pigs suggest that in vivo models may be more useful for further studies investigating the mechanisms underlying the cardiotoxicity of halofantrine. PMID- 9351517 TI - Acute gentamicin-induced hypercalciuria and hypermagnesiuria in the rat: dose response relationship and role of renal tubular injury. AB - 1. Standard renal clearance techniques were used to assess the dose-response relationship between acute gentamicin infusion and the magnitude of hypercalciuria and hypermagnesiuria in the anaesthetized Sprague-Dawley rat. Also investigated were whether these effects occurred independently of renal tubular cell injury. 2. Acute gentamicin infusion was associated with a significant hypercalciuria and hypermagnesiuria evident within 30 min of drug infusion. The magnitude of these responses was related to the dose of drug infused (0.14-1.12 mg kg(-1) min[-1]). Increased urinary electrolyte losses resulted from a decreased tubular reabsorption of calcium and magnesium. 3. A rapid dose-related increase in urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion was also observed in response to gentamicin infusion. However, there was no evidence of renal tubular cell injury and no myeloid bodies were observed within the lysosomes of the proximal tubular cells. Gentamicin may thus interfere with the mechanisms for cellular uptake and intracellular processing of NAG causing increased NAG release into the tubular lumen. 4. The absence of changes in renal cellular morphology indicates that the excessive renal losses of calcium and magnesium were an effect of gentamicin per se and not the result of underlying renal tubular injury. The renal effects described in this paper were apparent after administration of relatively low total drug doses, and with plasma concentrations calculated to be within the clinical range. These findings suggest that disturbances of plasma electrolyte homeostasis could occur in the absence of overt renal injury in patients receiving aminoglycoside antibiotics. PMID- 9351518 TI - Nitroglycerin-inhibited whole blood aggregation is partially mediated by calcitonin gene-related peptide -- a neurogenic mechanism. AB - 1. The role of the vasculature and calcitonin gene-related peptide (CGRP) in nitroglycerin (NTG)-mediated platelet inhibition was studied. 2. In vitro incubations of CGRP in whole blood induced a dose-dependent inhibition of platelet aggregation with an IC50 of 62.1 nM. 3. The platelet inhibition induced by CGRP was blocked by co-incubation of 0.53 microM CGRP8-37, as well as 30 microM N(G)-nitro-monomethyl-L-arginine (L-NMMA). 4. In a separate group of experiments, 100 nM NTG in rat whole blood (WB) induced platelet inhibition of 6.0 +/- 1.3% (mean +/- s.d.), which was enhanced to 77.6+/-3.5% by the addition of rat aortic tissue (AT) (P<0.001). The inclusion of CGRP8-37 with NTG and AT in WB reduced platelet inhibition to 31.6+6.8% (P<0.01). Incubation of WB and AT with 30 microM L-NMMA reduced NTG-induced inhibition of platelet aggregation to 26.4+/-4.2% (P<0.001). 5. It is concluded that vascular tissue contributes to the antiplatelet mechanism of action of NTG. Furthermore, NTG apparently evokes the release of CGRP from vascular tissue and this neuropeptide contributes to the antiplatelet actions of NTG. 6. The antiplatelet activity of CGRP in whole blood is mediated primarily through the activation of nitric oxide synthase. PMID- 9351520 TI - A toxin from the spider Phoneutria nigriventer that blocks calcium channels coupled to exocytosis. AB - 1. The aim of the present experiments was to investigate the pharmacological action of a toxin from the spider Phoneutria nigriventer, Tx3-3, on the function of calcium channels that control exocytosis of synaptic vesicles. 2. Tx3-3, in confirmation of previous work, diminished the intracellular calcium increase induced by membrane depolarization with KCl (25 mM) in rat cerebrocortical synaptosomes. The toxin was very potent (IC50 0.9 nM) at inhibiting calcium channels that regulate calcium entry in synaptosomes. In addition, Tx3-3 blocked the exocytosis of synaptic vesicles, as measured with the fluorescent dye FM1-43. 3. Using omega-toxins that interact selectively with distinct neuronal calcium channels, we investigated whether the target of Tx3-3 overlaps with known channels that mediate exocytosis. The results indicate that the main population of voltage-sensitive calcium channels altered by Tx3-3 can also be inhibited by omega-agatoxin IVA, an antagonist of P/Q calcium channels. Omega-conotoxin GVIA, which inhibits N type calcium channels did not decrease significantly the entry of calcium or exocytosis of synaptic vesicles in depolarized synaptosomes. 4. It is concluded that Tx3-3 potently inhibits omega-agatoxin IVA-sensitive calcium channels, which are involved in controlling exocytosis in rat brain cortical synaptosomes. PMID- 9351519 TI - Selective antagonism of the GABA(A) receptor by ciprofloxacin and biphenylacetic acid. AB - 1. Previous studies have shown that ciprofloxacin and biphenylacetic acid (BPAA) synergistically inhibit y-aminobutyric acid (GABA)A receptors. In the present study, we have investigated the actions of these two drugs on other neuronal ligand-gated ion channels. 2. Agonist-evoked depolarizations were recorded from rat vagus and optic nerves in vitro by use of an extracellular recording technique. 3. GABA (50 microM)-evoked responses, in the vagus nerve in vitro, were inhibited by bicuculline (0.3-10 microM) and picrotoxin (0.3-10 microM), with IC50 values and 95% confidence intervals (CI) of 1.2 microM (1.1-1.4) and 3.6 microM (3.0-4.3), respectively, and were potentiated by sodium pentobarbitone (30 microM) and diazepam (1 microM) to (mean+/-s.e.mean) 168+/-18% and 117+/-4% of control, respectively. 5-Hydroxytryptamine (5-HT; 0.5 microM)-evoked responses were inhibited by MDL 72222 (1 microM) to 10+/-4% of control; DMPP (10 microM) evoked responses were inhibited by hexamethonium (100 microM) to 12+/-5% of control, and alphabetaMeATP (30 microM)-evoked responses were inhibited by PPADS (10 microM) to 21+/-5% of control. Together, these data are consistent with activation of GABA(A), 5-HT3, nicotinic ACh and P2X receptors, respectively. 4 Ciprofloxacin (10-3000 microM) inhibited GABA(A)-mediated responses in the vagus nerve with an IC50 (and 95% CI) of 202 microM (148-275). BPAA (1-1000 microM) had little or no effect on the GABA(A)-mediated response but concentration dependently potentiated the effects of ciprofloxacin by up to 33,000 times. 5. Responses mediated by 5-HT3, nicotinic ACh and P2X receptors in the vagus nerve and strychnine-sensitive glycine receptors in the optic nerve were little or unaffected by ciprofloxacin (100 microM), BPAA (100 microM) or the combination of these drugs (both at 100 microM). 6. GABA (1 mM)-evoked responses in the optic nerve were inhibited by bicuculline with an IC50 of 3.6 microM (2.8-4.5), a value not significantly different from that determined in the vagus nerve. Ciprofloxacin also inhibited the GABA-evoked response with an IC50 of 334 microM (256-437) and BPAA (100 microM) potentiated these antagonist effects. However, the magnitude of the synergy was 48 times less than that seen in the vagus nerve. 7. These data indicate that ciprofloxacin and BPAA are selective antagonists of GABA(A) receptors, an action that may contribute to their excitatory effects in vivo. Additionally, our data suggest that the molecular properties of GABA(A) receptors in different regions of the CNS influence the extent to which these drugs synergistically inhibit the GABA(A) receptor. PMID- 9351521 TI - Evaluation of an automated photometric fibrinogen assay. AB - A recently-introduced automated method for the determination of plasma fibrinogen is based on the principle of von Clauss, combined with photometric detection: after addition of thrombin, the coagulation time is determined by measuring the change in absorption at 405 nm. This method was evaluated and compared with the original coagulometric Clauss assay and with the prothrombin time (PT)-derived automated method. The inter-assay coefficient of variation of the Clauss-derived assay was lower (14.1, 3.8 and 4.6%) than the PT-derived assay (16.1, 7.5 and 10.5%, respectively) at all three fibrinogen levels tested (1.2, 4.0 and 7.5 g/l). The correlation between the assays was investigated according to the method of Passing and Bablok and could be described as follows: Clauss-derived = 0.79 (PT-derived) + 0.66; Clauss-derived = 1.12 (Clauss) + 0.143. The interference of heparin (< 1.5 U/ml), haemoglobin (< 30 micromol/l), bilirubin (< 200 micromol/l) and triglycerides (< 5.5 mmol/l) in the Clauss-derived assay was negligible. The effects of fibrinogen degradation products on the Clauss-derived assay were comparable with the effects on the Clauss assay, in contrast to the effects on the PT-derived assay. In conclusion, the Clauss-derived assay is a specific and precise automated method to determine fibrinogen concentrations in plasma, which is not liable to interference from different pathophysiological substances. PMID- 9351522 TI - A Protac-based screening test for activated protein C-resistant factor Va and other defects of the protein C anticoagulant pathway. AB - We have standardized a simple screening test for abnormalities of the protein C anticoagulant system. The test is basically a modified prothrombin time in which one aliquot of a test plasma is incubated for 3 min at 37 degrees C with Protac and another is incubated with buffer. During the incubation the Protac activates both protein C and factor V. The plasmas are then clotted with thromboplastin plus Ca2+, and the clotting time difference reflects the ability of the activated protein C (APC) to inactivate factor Va. With the use of Thromboplastin C Plus as the activator, clotting time differences found in 31 normal subjects (10.4 +/- 3.5 s, mean +/- 2SD) were distinct from clotting time differences found in 57 of 58 subjects with established APC-resistant factor Va (3.6 +/- 3.0 s). In addition, the Protac-based test detected six of seven patients with isolated protein C deficiency and 20 of 28 patients with isolated protein S deficiency. Because of the reported high prevalence of heterozygous APC-resistant factor Va in Caucasian populations, it should be particularly useful in determining whether this genetic risk is present in individuals who have experienced or are at increased environmental risk of venous thrombosis. PMID- 9351523 TI - Molecular analysis of a compound heterozygote for hypoprothrombinemia and dysprothrombinemia (-G 7248/7249 and ARG 340 TRP). AB - Hypoprothrombinemia is an uncommon hereditary coagulation defect characterized by low levels of biologically active prothrombin. Automated fluorescence-based DNA sequence analysis of amplified genomic DNA was used to define prothrombin gene regions from a patient with severe functional hypoprothrombinemia and little detectable prothrombin antigen. Two changes that alter amino acid sequence were observed: a deletion of one nucleotide (-G, 7248/7249) in exon 8 of one allele, causing a frameshift at codon 249/250 that results in premature termination of translation; and a C --> T change resulting in the substitution of tryptophan (TGG) for arginine (CGG) at amino acid 340 in exon 10 of the prothrombin gene. Computer modeling of the thrombin molecule confirmed that arginine 340 is located at the surface of the thrombin molecule, which points to the aqueous solvent. As tryptophan is a highly hydrophobic amino acid, the Arg --> Trp change may be associated with instability of the thrombin molecule. PMID- 9351524 TI - Increased serum levels of thrombopoietin in patients with thrombotic thrombocytopenic purpura, idiopathic thrombocytopenic purpura, or disseminated intravascular coagulation. AB - The serum levels of thrombopoietin (TPO) were measured in 16 patients with thrombotic thrombocytopenic purpura (TTP), 12 with hemolytic uremic syndrome (HUS), 10 with aplastic anemia (AA), 10 with disseminated intravascular coagulation (DIC), and 71 with idiopathic thrombocytopenic purpura (ITP). The serum TPO levels were measured with a sensitive sandwich enzyme-linked immunosorbent assay. The serum TPO level in the ITP group (1.68 +/- 0.85 fmol/ml) were not significantly increased compared with those of the normal subjects. The TPO levels in the TTP (2.77 +/- 1.38 fmol/ml) and HUS groups (5.77 +/- 4.41 fmol/ml) were higher than those of the normal subjects. The patients with AA (12.7 +/- 8.0 fmol/ml) and those with DIC (13.3 +/- 5.7 mol/ml) had significantly higher serum TPO levels than did the normal subjects and ITP patients. The TPO levels were well correlated with the platelet counts in the TTP patients, and were negatively correlated with the platelet counts in the ITP patients. These results suggest that the serum TPO levels in some thrombocytopenic diseases are regulated not only by the platelet count and the megakaryocyte mass, but also by other factors. PMID- 9351526 TI - APC resistance and factor V Leiden (FV:Q506) mutation in patients with ischemic cerebral events. Vienna Thrombophilia in Stroke Study Group (VITISS) PMID- 9351525 TI - Rheological study of the dynamic process of fibrinolysis. AB - The dynamic process of fibrinolysis induced by tissue plasminogen activator was examined using a rheological technique. Change in a rheological parameter (logarithmic damping factor) of whole blood and platelet-free plasma during fibrinolysis was largely dependent on the initial concentration of tissue plasminogen activator. Addition of activated partial prothrombin time reagent allowed determination of the time both of onset and end of fibrinolysis without affecting the coagulation process. The changes in the clot structure of fibrin during fibrinolysis were observed with a scanning electron microscope and compared with the time-dependent behavior of the logarithmic damping factor. Differences in the logarithmic damping factor during fibrinolysis were evident in alteration of the network structure of clots. It will be shown that the present rheological technique is useful for examining the concentration dependence of fibrinolytic reagent on fibrinolysis as well as for monitoring the dynamic process of fibrinolysis. PMID- 9351527 TI - Exogenous heparin reduces soluble plasma thrombomodulin levels. PMID- 9351528 TI - Relationship between factor VIII replacement therapy and joint damage in severe haemophilia. AB - Most of the physical, psychosocial and financial disability in severe haemophilia A is caused by the effects of recurrent haemarthroses and of chronic arthritis. Recent evidence suggests that this morbidity is related to insufficient factor VIII replacement therapy, not only quantitative (annual dose) but also qualitative (need for regular long-term prophylaxis). The recent development of recombinant factor VIII may increase the acceptability of prophylaxis to parents and patients, and hence reduce morbid outcomes. PMID- 9351530 TI - Central venous access devices in children with hemophilia: an update. AB - The most frequent indication for placement of a central venous access device in hemophiliacs is in very young boys (ages 1-2 years) with severe hemophilia who are started on a program of long-term factor prophylaxis designed to eliminate target joint bleeding and the development of chronic musculoskeletal disease. Although expensive, this strategy is extremely successful. It involves intravenous infusion of 25-40 factor units per kg on alternate days (minimum 3 times a week) for boys with severe hemophilia A, and twice a week for boys with severe hemophilia B. To facilitate this prophylaxis regimen some hemophilia clinics routinely recommend placement of a central venous access device; others, more concerned about associated complications such as sepsis, stress the importance of using peripheral veins wherever possible, with central access devices reserved for occasional, selected cases only. A decision to use such a device should only be made after discussion of the risks/benefits with parents (or guardians) and with patients if of an appropriate age. If such a system is to be used, we recommend that a totally implantable device (Port-A-Cath) be placed because of the lower risk of infection, and because totally implantable devices allow children to take part in activities such as swimming. Important complications include catheter-related sepsis, which may occur in 25% or more of devices over time and, much less frequently, catheter-related deep vein thrombosis. PMID- 9351529 TI - Prophylaxis in haemophilic children. AB - Prophylaxis with recombinant factor concentrates is the standard for children with severe haemophilia at the end of the 1990s but there are still many who are not given this optimal therapy for a variety of reasons; the dominant one globally is almost certainly financial, and efforts over the next decade must concentrate on ways of reducing costs and reducing factor concentrate requirements. Dosing according to kinetic principles can give a more cost effective use of concentrate and a computerized pharmacokinetic model can be used. Another possibility is the introduction of an implantable sustained-release pump delivery system connected to a central venous access system, which can deliver factor concentrate to maintain a constant level of around 5% and thus reduce the overall amount of factor used. The dose of factor required with this system has been estimated as 700-875 IU/kg per year to keep the plasma level at 2% and 1700-2200 IU/kg per year to maintain it at 5%. PMID- 9351531 TI - Acquired hemophilia and its treatment. AB - The development of auto-antibodies against clotting factor VIII is a rare disease that occurs predominantly in adults: in pregnant women or people with various immunologic disorders or with no apparent underlying disease. As a consequence of an immune system dysregulation, the evolution of this condition is unpredictable. The aim of treatment is to restore normal factor VIII levels in circulation using desmopressin (DDAVP), massive doses of factor VIII (human or porcine), plasmapheresis and factor VIII infusions. In patients with high-titer inhibitors, products with 'bypasing' activity can be used. The most difficult task is to treat the immune disorder. High-dose infusions of immunoglobulins might be useful, due to the presence of anti-idiotypes to factor VIII inhibitors in the commercial preparations. Corticosteroids and cyclophosphamide are currently the most widely used treatments for the immune disease. PMID- 9351532 TI - Viral safety of plasma-derived factor VIII and IX concentrates. AB - Treatment, or prevention, of bleeding in severe haemophilia requires lifelong therapy with repeated injections of factor VIII/IX concentrate which carry a risk of viral infection. The overall safety of all blood products depends not only upon careful donor selection and the screening of donations for infectious viruses, but also on the efficacy of specific anti-viral steps in the manufacturing process. As a result of the episodes of virus transmission by currently available concentrates there have been calls for all manufacturing processes to include two viral inactivation procedures. There have been no clinical studies, however, to demonstrate that these products have enhanced viral safety. A variety of other virucidal techniques are under evaluation. The way forward may be a form of post-marketing surveillance or pharmacovigilance in which large numbers of haemophiliacs are routinely screened for known transmissible viruses and the data systematically collected. It would probably be necessary to have a multinational arrangement because of the rarity of the transmission episodes. Such a system could also provide quick early warning of a difficulty with an individual product. PMID- 9351533 TI - Who should receive recombinant factor VIII? AB - Recent guidelines published by the United Kingdom Haemophilia Centre Directors' Organisation state that recombinant factor VIII constitutes the treatment of choice for all patients with haemophilia A. However, certain categories may have to be accorded priority as financial resources are limited. The general order of priority for the introduction of recombinant factor VIII is (1) previously untreated patients, (2) human immunodeficiency virus (HIV)- and hepatitis C negative patients, (3) HIV-negative, hepatitis C-positive patients and (4) patients seropositive for anti-HIV. PMID- 9351534 TI - Previously untreated patients and recombinant factor VIII concentrate studies. AB - Inhibitors are more common than previously thought in any population of patients with haemophilia, particularly those with severe haemophilia. Comparison of a recent analysis of retrospective data on inhibitor incidence with data derived from recombinant factor VIII studies suggests that the incidence of inhibitors is not substantially different between the two groups. However, a distinction must be made between high- and low-responding inhibitors and it is not clear whether low-level inhibitors in some patients are part of the natural history of haemophilia. Questions remain on why some groups might differ in inhibitor levels. Further work is required to ensure standardization of future protocols and continued observation of present cohorts. PMID- 9351535 TI - Recombinant factor VIII in hemophilia A: the Canadian experience. AB - Programs that provide health care for people with inherited bleeding disorders should strive to ensure that all patients have equal access to the safest and most efficacious replacement products. The widespread introduction of very high purity, plasma-derived and recombinant factor concentrates for use in the Canadian hemophilia population in October 1993 is consistent with this goal. The data from Canada are reassuring in that the prevalence of inhibitors has not increased significantly in a large number of severe hemophilia A patients switched to recombinant factor VIII concentrate, and that no cases of therapy associated thromboses have been reported with hemophilia B patients switched to high purity factor IX concentrates. However, high-purity factor concentrates are very expensive. In Canada, funding for health care is a government-based responsibility. In other countries where funding for health care is organized differently, other treatment recommendations may be more appropriate. PMID- 9351536 TI - Pharmacoeconomics of haemophilia. AB - Pharmacoeconomic analysis is one method of providing health-care decision-makers with information to assist them in assigning priorities for the allocation of resources, both within a particular disease area and between different diseases. Conducted well, such analyses can provide a valuable contribution to the published literature which can contribute to improving clinical outcomes through the endorsement of evidence-based medicine. The treatment of haemophilia with replacement therapy is an area which could benefit from such an approach, both by modelling pharmacoeconomic analyses on published clinical studies and by incorporating pharmacoeconomic analysis and quality of life measurements alongside existing clinical trials. PMID- 9351537 TI - Histologic grade as a prognostic factor in breast carcinoma. PMID- 9351538 TI - The role of histologic grading in the prognosis of patients with carcinoma of the breast: is this a neglected opportunity? AB - BACKGROUND: Quantified grading of breast carcinoma through histologic analysis has been practiced for many years, but generally has not been used as an aid in treatment decision-making. METHODS: A representative sample of the literature is reviewed and discussed. RESULTS: The literature overwhelmingly confirms that histologic features suitable for grading provide important prognostic information at all stages of the disease, but there is no uniformly agreed on methodology for the application of this information. Disagreement still exists as to its validity and it is often ignored. CONCLUSIONS: It appears reasonable that serious attempts be made to overcome the perceived problems with the grading of breast carcinoma and to develop a consensus regarding a methodology for quantifying this well established index of the virulence of the disease that is comparable to the consensus reached in the American Joint Committee on Cancer staging of the extent of disease. This information then could be used in the design of clinical trials. [See editorial counterpoint on pages 1703-5 and reply to counterpoint on pages 1706-7, this issue.] PMID- 9351539 TI - Early response to therapy and outcome in childhood acute lymphoblastic leukemia: a review. AB - BACKGROUND: Early response to therapy is defined as the initial response prior to Day 28 of treatment, the conventional time of marrow evaluation. The number of reports linking early response to therapy with the ultimate outcome of childhood acute lymphoblastic leukemia is substantial and growing. When this study began, these experiences had yet to be comprehensively reviewed. METHODS: A comprehensive search of the published literature yielded contributory reports of 14 trials conducted in the United States and Europe. In addition, unpublished data from one Children's Cancer Group trial were made available. Outcome measures were standardized by conversion to ratios of the incidence of adverse events among poorer and better responders. RESULTS: Early response to therapy was an independent prognostic factor in each of the 15 trials, which together included more than 10,000 patients. The incidence of slower early response ranged from 2 33%, with various measures and criteria used in different trials. Patients with a slower early response were 1.5-6.1 times (median, 2.7) more likely to have an adverse event than patients with a more rapid early response, however defined. Early response maintained prognostic significance after the exclusion of induction failure and within risk strata defined by age, white blood cell count, and/or immunophenotype. Its significance was also maintained in multivariate analyses where performed. CONCLUSIONS: Early response to therapy, whether determined by evaluation of bone marrow or peripheral blood, is a consistent, independent prognostic factor in childhood acute lymphoblastic leukemia. Slower early response may serve as a useful surrogate for outcome, a more complex end point, in investigations of the cellular and molecular determinants of resistance to therapy. It may also allow early identification of a patient subpopulation for whom current therapy is less effective and alternative strategies may be justified. PMID- 9351541 TI - Low serum levels of soluble CD44 variant 6 are significantly associated with poor prognosis in patients with pancreatic carcinoma. AB - BACKGROUND: Variant CD44 splice products, especially CD44 variant 6 (CD44v6), are expressed on activated lymphocytes and tumor cells. The soluble forms of CD44 standard (CD44s) and CD44v6 are present in the serum of normal individuals. The aim of the current study was to evaluate the concentrations and the prognostic potential of soluble CD44s and CD44v6 in patients with pancreatic carcinoma. METHODS: The serum CD44s and CD44v6 levels were determined quantitatively by enzyme-linked immunosorbent assay. The molecular mass of CD44v6 isoforms was determined by immunoprecipitation and Western blot analysis. CD44 mRNAs were analyzed by reverse transcriptase polymerase chain reaction followed by exon specific analysis. RESULTS: Both serum CD44s and serum CD44v6 were significantly reduced in patients with pancreatic carcinoma (n = 93, P < 0.001 and P < 0.00005). The median survival in the group with CD44v6 serum concentrations below 100 ng/mL was significantly decreased compared with that in the group with serum concentrations higher than 100 ng/mL (6.7 vs. 15.1 months, P < 0.0005). The isoforms containing soluble CD44v6 (sCD44v6) that were detected in the sera of pancreatic carcinoma patients showed molecular masses comparable to the sCD44v6 isoforms detected in the supernatant of lymphocytes activated by phorbol myestral acetate, whereas the sCD44v6 isoforms detected in the supernatant of pancreatic carcinoma cell lines exhibited higher molecular masses. CONCLUSIONS: These results suggest that serum CD44v6 is significantly reduced in pancreatic carcinoma patients and could serve as a good prognostic marker for patients with this disease. PMID- 9351540 TI - 9-Aminocamptothecin by 72-hour continuous intravenous infusion is inactive in the treatment of patients with 5-fluorouracil-refractory colorectal carcinoma. AB - BACKGROUND: 9-Aminocamptothecin (9AC) and its parent compound, camptothecin, have shown outstanding preclinical activity against colorectal carcinoma. Irinotecan (CPT-11), another camptothecin derivative, has demonstrated clinical activity in patients with 5-fluorouracil (5-FU)-refractory colorectal carcinoma. METHODS: The authors performed a Phase II trial of 9AC involving patients with measurable metastatic colorectal carcinoma who had progressed through only one prior regimen of 5-FU-based chemotherapy. 9AC was given initially at a dose of 59 microg/m2/hour by continuous intravenous infusion for 72 hours, with treatments repeated every 14 days. Granulocyte-colony stimulating factor was given on Days 5 12. RESULTS: Sixteen patients were treated on this trial. Fourteen were evaluable for response. Contrary to expectations, no major objective antitumor responses were observed. Eight patients experienced stable disease for a median of 4.1 months (range, 2.2-9.5 months). Toxicities, especially myelosuppression, were severe and necessitated a 15% reduction in the initial dose after the first 9 patients. Toxicities at this reduced dose remained unacceptable. CONCLUSIONS: 9AC did not demonstrate substantial activity against 5-FU-refractory colorectal carcinoma on the schedule studied. Toxicities at the doses and schedule studied were unacceptable in this patient population. Based on their results, the authors consider it unlikely that 9AC administered as a 72-hour continuous intravenous infusion will play a major role in the treatment of colorectal carcinoma. PMID- 9351542 TI - The clinical behavior of breast carcinoma is probably determined at the preinvasive stage (ductal carcinoma in situ). AB - BACKGROUND: Mammography has greatly increased the number of women diagnosed with ductal carcinoma in situ (DCIS) of the breast. New classifications of DCIS recently have been proposed. In this study, these classifications were applied to DCIS associated with invasive tumors and correlated with patient prognosis. METHODS: Three hundred cases of infiltrating ductal carcinoma of the breast in which there was associated DCIS in the surrounding breast were studied. The DCIS was classified as well, moderately, or poorly differentiated, according to two recently published systems using the cytonuclear features of the malignant cells (Holland classification) and cytology and the presence of necrosis (Van Nuys classification). RESULTS: The differentiation of DCIS was significantly correlated with the grade of the invasive carcinoma (P < 0.0001), the Nottingham prognostic index (P < 0.0001), and the clinical outcomes of the patients (P < 0.0001). CONCLUSIONS: These findings indicate that a model in which there is increasing in situ dysplasia before the development of stromal invasion is incorrect for breast carcinoma. Rather, in most cases, well-differentiated DCIS probably gives rise to low grade invasive breast carcinoma with a better long term clinical outcome. These data suggest that many of the important prognostic biologic and genetic characteristics of breast carcinoma are already established in the neoplastic clones of malignant cells at the preinvasive stage of the disease. PMID- 9351543 TI - Surgical treatment of 70 patients with brain metastases from breast carcinoma. AB - BACKGROUND: Brain metastases are diagnosed in 15% of patients with metastatic breast carcinoma. Most patients are treated with whole-brain radiotherapy (WBRT) and/or chemotherapy. The information on surgical results is sparse. METHODS: Among 709 patients with tumors metastatic to the brain who underwent craniotomy at Memorial Hospital, New York, New York, between January 1974 and December 1993, 70 (10%) had a primary breast carcinoma. Their treatment outcomes were analyzed retrospectively. RESULTS: The median age at diagnosis of primary breast carcinoma and brain metastasis was 46 and 50 years, respectively. All but two patients had metachronous diagnoses of breast carcinoma and brain metastasis. The median interval between both diagnoses in this subgroup was 28 months. In all 70 patients, the overall median survival was 54 months after diagnosis of the primary breast tumor and 16.2 months after diagnosis of the brain tumor. Only 5 patients (7%) were alive at last follow-up. The overall median survival after brain surgery was 14 months. Four patients died within 30 days of craniotomy. Twelve patients had a solitary cerebellar metastasis and 16 had multiple metastases; their median survival was 10.9 months and 14.8 months, respectively. There was no statistical difference in survival for patients who had single or multiple lesions. The median survival of 22 patients with positive hormonal receptor (estrogen receptor [ER] or progesterone receptor [PR]) was significantly longer than the median survival of 20 patients with negative ER/PR (21.9 vs. 12.5 months, P < 0.05). For 35 patients (50%) who had brain lesions > or =4 cm, the median survival was 11 months, compared with 16.3 months for patients with smaller lesions (P = 0.16, not significant [NS]). Patients age < or =50 years versus >50 years had survival of 17.3 and 11.1 months, respectively (P = NS). Neurologic deficit prior to craniotomy shortened survival for 24 patients to 11.5 months, compared with 17.4 months for patients without deficit (P = NS). Fifteen patients experienced failure with WBRT prior to undergoing craniotomy, and their median survival was shorter than for those who underwent craniotomy as the initial treatment (6.3 vs. 15.8 months, P < 0.03). However, their survival after diagnosis of brain metastasis was not significantly different (19.2 vs. 16.1). Forty-seven patients received WBRT postoperatively, and 9 patients did not receive adjuvant radiation therapy. Subsequent relapse in the brain was diagnosed in 27 patients, and 8 of them underwent reresection. One-year, 2-year, 3-year, and 5-year survival rates were 53%, 25.7%, 18.6%, and 7%, respectively. In multivariate analysis, the adjuvant WBRT after craniotomy and the absence of meningeal carcinomatosis were the only significant predictive variables for longer survival. CONCLUSIONS: In a subset of selected patients, craniotomy followed by WBRT can positively impact survival. PMID- 9351544 TI - Simultaneous antiandrogen withdrawal and treatment with ketoconazole and hydrocortisone in patients with advanced prostate carcinoma. AB - BACKGROUND: Although antiandrogen withdrawal has moderate efficacy in patients with hormone refractory prostate carcinoma (HRPC), the effect of the simultaneous suppression of adrenal androgens with ketoconazole at the time of antiandrogen withdrawal is not known. METHODS: Twenty consecutive patients with HRPC who had developed progressive disease despite combined androgen blockade were treated with antiandrogen withdrawal and simultaneous ketoconazole as a means of inhibiting adrenal steroid production. Prostate specific antigen (PSA) response was defined as a > 50% fall in PSA from baseline that was maintained for at least 8 weeks. RESULTS: Ten patients had established metastatic disease, 2 had high PSAs and no imaging studies (PSA of 70 and 160 ng/mL, respectively), 3 had microscopically positive lymph nodes and serologic progression, and 5 had serologic progression alone. Overall, of 20 evaluable patients, 11 (55%) had a > 50% fall in PSA (95% confidence interval [CI], 31.5-76.9%). The median PSA response duration was 8.5 months (95% CI, 7-17 months). The median survival was 19 months. Toxicity was mild, with Grade 1 and 2 nausea and emesis in 15% of patients, Grade 1 fatigue in 10% of patients, and reversible Grade 1 or 2 hepatotoxicity in 10% of patients. Mild skin toxicity was observed in 20% of patients. CONCLUSIONS: The addition of ketoconazole and hydrocortisone to antiandrogen withdrawal appears to increase the PSA response proportion observed with antiandrogen withdrawal alone. Toxicity is mild. PMID- 9351545 TI - Serum concentration of type I collagen metabolites as a quantitative marker of bone metastases in patients with prostate carcinoma. AB - BACKGROUND: Bone scans, widely used for the detection of bone metastases from prostate carcinoma, can neither quantitate metastatic lesions nor detect osteolytic lesions. METHODS: Serum concentrations of the carboxyterminal propeptide of Type I procollagen (PICP), the carboxyterminal pyridinoline cross linked telopeptide of Type I collagen (ICTP), and prostate specific antigen (PSA) were measured by radioimmunoassays in 48 patients with benign prostatic hyperplasia (BPH), 25 patients with prostate carcinoma (PCA) without bone metastases, and 36 patients with PCA and bone metastases. RESULTS: Serum concentrations of PICP were significantly higher in patients with PCA with bone metastases than in patients with BPH or PCA without bone metastases. No significant differences were observed between patients with BPH and those with PCA without bone metastases. Serum ICTP concentrations were significantly higher in patients with PCA than in patients with BPH regardless of the presence or absence of bone metastases. Serum concentrations of PICP, ICTP, and PSA correlated significantly with Soloway's grading system for bone scans. The serum concentrations of PICP and ICTP in patients without bone metastases showed a significant downward trend in response to antiandrogen therapy. CONCLUSIONS: These observations suggest that the serum concentrations of PICP and ICTP are quantitative markers of bone metastases from PCA when followed serially in individual patients. PMID- 9351546 TI - Appraisal of intratumoral microvessel density, MIB-1 score, DNA content, and p53 protein expression as prognostic indicators in patients with locally confined renal cell carcinoma. AB - BACKGROUND: The prognostic values of intratumoral microvessel density (iMVD), tumor cell proliferation rate, DNA content (ploidy), and p53 protein expression are controversial or have not been well studied in patients with renal cell carcinoma (RCC) confined to the kidney. METHODS: A uniform group of 52 clear cell (conventional) RCCs confined to the kidney (classified as T1N0M0 or T2N0M0) were analyzed for iMVD, MIB-1 score, DNA content, S-phase fraction, and p53 protein expression by immunohistochemical methods or flow cytometry. iMVD was evaluated in a single area (X200, 1.15 mm2) representative of the highest MVD (neovascular "hot spot") after independently highlighting endothelial cells with antibodies specific for factor VIII-related antigen (F8/86) and CD31 (JC/70A). The MIB-1 antibody (Ki-67 antigen) score was used as a marker for the tumor cell proliferation rate. DNA content and S-phase fraction were determined by flow cytometry using paraffin embedded tissue. p53 expression was assessed using the D07 antibody. RESULTS: The median time of clinical follow-up was > 9 years. Eleven patients died of disease; the median time to death was 26 months. iMVD counts using antifactor VIII and anti-CD31 were tightly correlated (correlation coefficient = 0.89). S-phase fraction was higher in aneuploid tumors than in diploid tumors (mean, 12.4% vs. 4.3%; P = 0.01). Using univariate survival analyses, tumor size (stage classification pT1 vs. PT2; P = 0.01) and nuclear grade (P = 0.04) were associated with shortened survival. No statistically significant differences in survival were found for iMVD, MIB-1 score, DNA content, S-phase fraction, or p53 expression. Only two cases strongly expressed p53 protein; both tumors were of high nuclear grade. Using multivariate survival analyses, nuclear grade and tumor size were the only independent prognostic factors (best model P = 0.002). CONCLUSIONS: In this study, nuclear grade and tumor size were found to be independent predictors of survival in locally confined clear cell (conventional) RCC, as has been shown previously for locally confined RCC in general. MIB-1 score, iMVD counts, DNA content, S-phase fraction, and p53 expression did not contribute additional prognostic information. PMID- 9351547 TI - Combined intraarterial chemotherapy and radiotherapy in the treatment of bladder carcinoma. AB - BACKGROUND: The combination of radiotherapy and cisplatin-based chemotherapy has proved to be an effective treatment for bladder carcinoma in many clinical studies. Intra-arterial approaches to chemotherapy have been developed to reduce systemic toxicities and improve response rates. This study was designed to determine the effectiveness of intra-arterial chemotherapy with cisplatin and doxorubicin combined with radiotherapy in the treatment of patients with invasive bladder carcinoma. The objectives were to evaluate the response rate, bladder preservation rate, toxicity, and survival rate. METHODS: Thirty-five patients with muscle-invasive bladder carcinoma at clinical stage T2-T4N0M0 were each treated with 2courses of intra-arterial cisplatin and doxorubicin at 3-week intervals, whereas radiotherapy was administered for 4 weeks (2 gray [Gy] given a total of 20 times, at 5 fractions per week). Patients with complete responses were given an additional course of chemotherapy (intra-arterial cisplatin and doxorubicin) and irradiation (20 Gy), and patients with residual tumor after the initial chemoradiotherapy underwent cystectomy. RESULTS: A clinical complete response was observed in 26 patients (74%; 95% confidence interval, 59-89%), and an incomplete response was observed in 9 (26%; 95% confidence interval, 11-41%). The bladder was preserved in all patients with a complete response, and it was tumor free in 19 of them (54% of all patients). The actuarial survival rate was 76.6% at 5 years. After a median follow-up interval of 45 months, 28 patients (80%) were alive and 7 (20%) had died due to disease progression. The regimen was well tolerated, with no severe systemic or local toxicities. CONCLUSIONS: The high rates of response, survival, and bladder preservation observed indicate that this combined intra-arterial chemotherapy and radiotherapy regimen would be useful in the management of invasive bladder carcinoma. This was a small Phase II trial; the results are preliminary, and the utility of this treatment modality in patient management remains to be proven. PMID- 9351548 TI - Langerhans' cell histiocytosis in adults: a clinical and therapeutic analysis of 11 patients from a single institution. AB - BACKGROUND: Langerhans' cell histiocytosis (LCH) is a rare disorder of uncertain etiology, characterized by a wide clinical spectrum and varied behavior. METHODS: This retrospective study analyzed 11 adult patients with a diagnosis of LCH observed at the study institution between April 1988 and March 1993. RESULTS: Based on the sites and extent of disease at diagnosis, patients were divided into four categories. Group A was comprised of four patients with unifocal bone disease who had surgical curettage. At last follow-up only 1 patient was in continuous complete response (CCR) at 29+ months. The other 3 patients recurred at 3, 12, and 30 months, respectively, after surgery and at last follow-up were found to be in CR at 16+, 48+, and 124+ months, respectively, after therapy with vinblastine (VBL) and high dose methylprednisolone (HDMP). Group B was comprised of three patients with multifocal bone disease. Two of these patients received VBL + HDMP; at last follow-up, 1 patient was in CCR 8 months after completion of therapy, and the other developed progressive disease 11 months later. The third patient was treated with interferon (IFN) and at last follow-up was in CCR at 35+ months. Group C was comprised of 2 patients with bone and visceral disease who were treated with etoposide (VP-16) + HDMP; at last follow-up, 1 patient was in CCR at 42+ months and the other patient, who had isolated vulvar recurrence 16 months later, was in CR with treatment with local IFN. Group D was comprised of two patients with lung and lymph node involvement, one of whom was treated with VP-16 + HDMP and the other with cyclophosphamide, doxorubicin, vincristine, and prednisone; at last follow-up, both were in CCR at 30+ and 71+ months, respectively. CONCLUSIONS: VBL + HDMP showed efficacy in patients with bone disease, in particular those treated for recurrent LCH after surgery. Therapy with VP-16 and HDMP was successfully employed in patients with visceral disease. IFN was effective both for localized disease and in patients with multiple bone lesions. PMID- 9351549 TI - Nonmetastatic intracranial germinoma: the experience of the French Society of Pediatric Oncology. AB - BACKGROUND: Standard treatment of localized intracranial germinoma is focal irradiation of the primary tumor (45-50 grays [Gy]) combined with craniospinal radiotherapy (RT). To decrease late effects related to extensive fields of RT, the French Society of Pediatric Oncology decided in 1990 to replace prophylactic RT with chemotherapy (CT) and to deliver focal RT at 40 Gy. METHODS: Twenty-nine patients with localized, biopsy proven germinoma were included in this study between January 1990 and December 1994. CT consisted of 2 cycles of carboplatin 600 mg/m2 on Day 1, etoposide 150 mg/m2 on Days 1-3, ifosfamide 1.8 g/m2 on Days 22-26, and etoposide 150 mg2 on Days 22-24, followed by RT delivered to the initial tumor volume (40 Gy). RESULTS: The median age of the 19 boys and 10 girls was 12.8 years; 25 patients had a unifocal tumor in the pineal (13), suprasellar (10), or thalamic (2) area, and 4 patients had a bifocal tumor. Three patients initially had complete surgery. Of the 26 patients evaluable for CT response, 11 had a small amount of tumor residue and 15 no residue; no patient underwent surgery after CT or RT. One patient recurred 3 years after diagnosis and is in his second complete remission. Twenty-eight patients are in their first complete remission after a median follow-up of 32 months (range, 7-68 months); 9 of the 28 have a small amount of tumor residue that is considered nonevolving. Overall survival at 4 years is 100% and event free survival is 93.3% (+/- 6%) after a median follow-up of 32 months. CONCLUSIONS: This treatment strategy avoids craniospinal RT and reduces focal RT, with results equivalent to those achieved with extensive RT. Thus, the authors consider it a valid treatment of nonmetastatic germinoma. PMID- 9351550 TI - Consensus Conference on the classification of ductal carcinoma in situ. The Consensus Conference Committee. PMID- 9351551 TI - TNM Classification of Malignant Tumors, fifth edition (1997). Union Internationale Contre le Cancer and the American Joint Committee on Cancer. PMID- 9351552 TI - American Cancer Society guidelines for the early detection of prostate cancer: update, June 10, 1997. PMID- 9351553 TI - An adjustment to the 1997 estimate for new prostate cancer cases. PMID- 9351554 TI - Observations on the early detection of prostate cancer from the American Cancer Society National Prostate Cancer Detection Project. AB - BACKGROUND: The American Cancer Society National Prostate Cancer Detection Project (ACS-NPCDP) was established in 1987. The experience of the ACS-NPCDP demonstrates the yield and impact of periodic examinations for the early detection of prostate cancer. METHODS: A cohort of 2999 well men ages 55-70 years was tested annually at 10 clinical centers by prostate specific antigen (PSA), transrectal ultrasound (TRUS), and digital rectal examination (DRE). Biopsies were performed on men with suspicious findings. Pathologic findings were reviewed. The initial study outcomes were the detection yield of multimodality testing and the comparative sensitivity and specificity of the different tests employed. Longer term outcomes included patient quality of life and survival. RESULTS: The cancer detection rate declined significantly across the years of intervention. DRE had lower sensitivity than TRUS or PSA, particularly in later years of follow-up. The specificity of TRUS was lower than that of DRE. Fewer than 9% of the cancers detected in this study were clinically advanced at the time of diagnosis. Ninety-four percent of patients in whom cancer was detected are alive after an average follow-up of 54 months. In one case, death occurred after surgery. Two deaths were attributed to prostate cancer, and eleven other deaths were unrelated to prostate cancer or its treatment. CONCLUSIONS: Results of the ACS-NPCDP indicate that a combined-modality approach to prostate cancer detection yields high levels of early detection with infrequent adverse outcomes. Continued follow-up is required to evaluate long term morbidity and mortality. PMID- 9351555 TI - Prostate carcinoma screening in the county of Tyrol, Austria: experience and results. AB - BACKGROUND: This article summarizes the experience and results of different prostate carcinoma screening projects using total prostate specific antigen (PSA) as the initial test and different diagnostic tests to improve specificity. METHODS: The seven projects studied included 1) a mass screening study using PSA as the initial test in 21,079 volunteers; 2) an investigation of the usefulness of normal and age-referenced PSA cutoffs in 1618 men; 3) a PSA-based screening study of 2272 asymptomatic blood donors; 4) an investigation of the incidence and clinical significance of transitional zone carcinoma in 340 men with negative rectal examination findings and clearly visible prostatic zones on three dimensional transrectal ultrasound; 5) determination of percent free PSA in one retrospective and two prospective screening studies to define the optimal range of total PSA and determine the appropriate cutpoints for percent free PSA within this range; 6) evaluation of the diagnostic benefit of PSA transitional zone density in 308 screening volunteers; and 7) a study of the impact of PSA-based screening on the percentage of incidental prostate carcinoma diagnosed in 1543 men undergoing transurethal resection of the prostate. RESULTS: 1) Of the 21,078 volunteers, 1618 (8%) had elevated PSA levels. Of these men, 778 (48%) underwent biopsies; 197 biopsies (25%) were positive for prostate carcinoma and 135 patients underwent radical prostatectomy. Ninety-five of the 135 pathologically staged lesions (70%) were found to be organ-confined. 2) A PSA cutoff of 2.5 ng/mL in men age 45-49 years and of 3.5 ng/mL in men age 50-59 years with normal digital rectal examination findings resulted in an 8% increase in both the number of biopsies (66 of 778) and the detection rate of organ-confined disease. 3) Of the 2272 men, 284 had elevated PSA levels and prostate carcinoma was detected in 62 men. All patients underwent radical prostatectomy and histologic examination revealed organ-confined disease in all but eight men. 4) Ninety-eight of 340 men (28.8%) had biopsies positive for carcinoma; 28 of these patients (28.5%) had carcinoma that originated in the transitional zone only. 5) In the retrospective study, receiver operating characteristic curve analysis showed that by using a percent free PSA of 18% as a biopsy criterion in men with an elevated PSA serum level, 37% of the negative biopsies could be eliminated although 94% of all carcinomas would still be detected. In the first prospective study, 106 of 158 men with elevated total PSA values between 2.5 and 10.0 ng/mL were further evaluated and 37 prostate carcinomas were detected. By using a percent free PSA of < or =22% as a biopsy criterion, 30% of the negative biopsies could be eliminated although 98% of the carcinomas would still be detected. In the second prospective study, 120 of 465 men with total PSA levels between 1.25 and 6.49 ng/mL, a percent free PSA of <18%, and normal digital rectal examination findings were further evaluated and 27 (22.5%) were found to have prostate carcinoma. 6) Receiver operating characteristic curve analysis for PSA transitional zone density showed that by using a PSA transitional zone density of >0.22 ng/mL/cc as a biopsy criterion, 24.4% of negative biopsies could be avoided without missing the detection of a single carcinoma. 7) In the prescreening era the incidence of T1a Grade 1 and 2 carcinomas was 3.1% and the incidence of T1a Grade 3 and T1b carcinoma was 2.3%, whereas in the years after the establishment of PSA-based screening the incidence was 4.6% and 1.03%, respectively. CONCLUSIONS: These data suggest that PSA-based screening increases the detection rate of clinically significant and organ-confined tumors. Percent free PSA and PSA transitional zone density provide an additional diagnostic benefit over total PSA. PMID- 9351556 TI - The European Randomized Study of Screening for Prostate Cancer: an update. AB - BACKGROUND: A consensus meeting on screening and global strategy for prostate carcinoma, held in Antwerp in 1994, determined the willingness among European cancer prevention centers to pursue vigorously the collaborative formation of a multinational randomized screening trial. This trial was to be named the European Randomized Study of Screening for Prostate Cancer (ERSPC). METHODS: During the years prior to that meeting, several feasibility trials were conducted in Antwerp and Rotterdam to evaluate the pitfalls and problems of a randomized procedure for population screening. Today, five centers in five European countries share their study work and results via the ERSPC, and others are lining up to join this massive effort. Regular meetings and specific work groups enable the research centers to compare their data, because the trial methodology differs slightly from one center to another. RESULTS: However, a common work strategy and analysis of the data has recently been reached, and the first study results of the trial (evaluating 180,000 men over a 10-year screening period) are expected by the year 2007. CONCLUSIONS: A randomized trial of prostate carcinoma screening is set up in Europe currently with five participating centers from five countries. First overall effect results of regular screening are expected after a 10-year period of follow-up. PMID- 9351557 TI - The results of prostate carcinoma screening in the U.S. as reflected in the surveillance, epidemiology, and end results program. AB - BACKGROUND: The rapid escalation in the incidence of prostate carcinoma between the years 1988 and 1992 has been attributed to prostate specific antigen screening. There have been concerns regarding the possible diagnosis and treatment of insignificant tumors in the absence of randomized, controlled trial evidence of a decrease in mortality. Descriptive studies suggest that serial screening decreases the detection of advanced disease. In November 1996, the National Center for Health Statistics recorded a decrease in prostate carcinoma mortality. METHODS: The basis of this analysis is 208,234 prostate carcinoma cases diagnosed between 1973 and 1993 in population-based Surveillance, Epidemiology, and End Results registries. The general staging system was used rather than that of the American Joint Committee on Cancer to permit observation of long term trends. Grade incorporating Gleason scores was used as an indication of the significance of the prostate carcinoma. Age-adjusted survival rates were used to separate prostate carcinoma deaths from deaths due to other causes. RESULTS: The increase in the incidence of prostate carcinoma has been greater than for any other malignancy. The increase was largely in Grade 2 significant tumors and not in Grade 1 (15%) insignificant tumors. There was a decrease in the detection of advanced disease. After the peak incidence in 1992, a progressive decrease to near baseline levels occurred. Approximately 38% of all deaths were from prostate carcinoma. Deaths from other causes increased with age. When corrected for death from other causes, men age > 69 years had a greater rate of death from prostate carcinoma than men age 50-69 years. Approximately 61% of all deaths from prostate carcinoma occurred within 5 years of diagnosis and 88% within 10 years. The 10-year survival rate for patients treated by radical prostatectomy was 100%, 78% for patients treated by radiation, and 33% for patients treated with other (noncomparable modalities). CONCLUSIONS: The indirect evidence suggested that prostate carcinoma screening of men ages >50 years decreased the incidence of distant disease, which influences the mortality rate. PMID- 9351558 TI - Eight years of "Prostate Cancer Awareness Week": lessons in screening and early detection. Prostate Cancer Education Council. AB - BACKGROUND: Prostate Cancer Awareness Week began in 1989 to raise public awareness of the disease and impact on its clinical dynamics. Prior to 1990, prostate carcinoma was usually diagnosed in symptomatic men as advanced and ipso facto incurable. METHODS: A 5-year longitudinal study of screening and early detection was initiated in 1992, involving 250 centers that tested over 50,000 men annually. The study analyzed the following: 1) the efficacy of digital rectal examination and prostate specific antigen (PSA) tests for the early detection of prostate carcinoma, 2) the impact of serial screening on stage of disease at diagnosis and cancer detection, 3) age and race specific reference ranges for PSA, 4) prostate carcinoma risk factors, and 5) psychosocial variables that influence the appropriateness and acceptability of both screening and treatment. RESULTS: Community-based screening for prostate carcinoma is as effective as programs conducted at academic centers, but annual testing may not be necessary for all men. Participants in community screening programs may require more prescreening information and education regarding the potential risks and benefits of screening, and also regarding the cascade of diagnostic and treatment decisions that follow an abnormal digital rectal examination or an elevated PSA finding. CONCLUSIONS: Community-based prostate carcinoma screening programs have contributed to the shift in the diagnosis of prostate carcinoma at an earlier stage. They provide data that are useful in studying the natural course of prostate carcinoma and in designing studies of the effect that prostate carcinoma screening has on mortality from the disease. PMID- 9351559 TI - The early detection of prostate carcinoma with prostate specific antigen: the Washington University experience. AB - BACKGROUND: It is not yet known whether screening for the detection of early prostate carcinoma will reduce mortality rates. However, data are available to assess intermediate outcomes from screening, including the performance characteristics of the screening tests and shifts in disease stage. METHODS: Approximately 30,000 community volunteers (mean age 60 years; <5% nonwhite) were enrolled in 1 of 3 screening studies. Volunteers were screened with PSA or PSA in combination with digital rectal examination at 6-month intervals, and prostatic biopsy was recommended for those with results suspicious for cancer. Based on a first-time screen, the current study reports screening test results, the proportion of men recommended to undergo biopsy, the proportion who actually underwent biopsy, and the carcinoma detection rates for each study, stratified by initial PSA level. The authors also report the pathologic features of screen detected carcinomas for a subset of men who underwent radical prostatectomy and for whom complete embedding and microscopic examination of the surgical specimen was performed. RESULTS: Approximately 10% of the volunteers had PSA levels >4.0 ng/mL and 3-10% had digital rectal examination results suspicious for cancer. Overall, 9-20% of volunteers were recommended to undergo biopsy and 8-13% actually underwent the procedure. The positive predictive value for carcinoma detection ranged from 25-33% across studies. In the subset of men for whom surgical specimens were completely embedded, the majority of tumors detected had the clinicopathologic features of significant carcinoma (<10% possibly harmless). CONCLUSIONS: The intermediate outcomes for screening with PSA and/or PSA in combination with digital rectal examination are encouraging. In community volunteers these screening tests demonstrated reasonable positive predictive value and detected carcinomas at an earlier stage. The majority of screen detected tumors had the pathologic characteristics of medically significant carcinoma. PMID- 9351560 TI - Prostate carcinoma incidence and patient mortality: the effects of screening and early detection. AB - BACKGROUND: Screening for and the aggressive treatment of prostate carcinoma are controversial, but they are nevertheless being practiced in the U.S. Current clinical studies of the effectiveness of screening will take years to complete. Meanwhile, screening for prostate carcinoma is already having an effect on society. METHODS: National and regional trends in prostate carcinoma incidence and data on patient mortality and survival from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute are described in this article. SEER is a population-based cancer data base comprised of nine discrete areas. Fundamental principles of screening are used in this article to explain the impact that prostate carcinoma screening has had in the U.S. RESULTS: According to the data in the SEER registries, overall prostate carcinoma incidence rates increased at a far greater pace than prostate carcinoma mortality rates during the period 1973-1994. During that period, there was a shift in stage at diagnosis characterized by an increase in local and regional disease, and a decline in distant disease at diagnosis. Overall 5-year survival rates for prostate carcinoma patients also increased. The increase in incidence rates, the shift in stage at diagnosis, and the increase in survival rates are all evidence of increasing early detection. However, these changes are consistent with lead time bias, length bias, a decline in mortality, and all three could have occurred. In the geographic SEER registries, the prostate carcinoma incidence rates vary markedly. These variations in incidence rates are due to regional variations in practice patterns and screening efforts. On the other hand, the SEER registries have comparable mortality rates. This is evidence of both lead time bias and length bias. CONCLUSIONS: Substantial regional variations in incidence were found, but regional mortality rates were similar. This is evidence that screening and early detection efforts are resulting in the diagnosis of prostate carcinoma in some men who do not need therapy; thus, prostate carcinoma screening can lead to unnecessary treatment for such men. Furthermore, epidemiologic data do not demonstrate that screening is decreasing mortality. The benefits of screening and early detection, although theoretically possible, are yet unproven, whereas the risks and harms of screening and resultant treatment are definite. PMID- 9351561 TI - Future benefits and cost-effectiveness of prostate carcinoma screening. American Cancer Society. AB - BACKGROUND: Estimates of cost-effectiveness for prostate carcinoma screening require a review of current data, modeling efforts, and perspectives on societal impact and costs. METHODS: Recent data from the cancer registry of the Michigan Department of Community Health was assessed for incidence trends in relation to age groups and racial differences. Differences in tumor biology between African American men (AAM) and white men were assessed from a large clinical biopsy series. A review of the literature addressing Markoff modeling to obtain estimates of treatment, screening efficacy, and costs were evaluated. RESULTS: The decline in the incidence of prostate carcinoma since 1992 primarily affected men age > 70 years whereas younger men (age 45-70 years) maintained a 100% greater incidence of localized disease than in 1989, when prostate specific antigen screening became more common. The rate of distant disease has decreased by 60% for both age groups. In the current biopsy series, AAM have distinctly more cores involved with carcinoma and have a higher number of carcinoma cores involved with a Gleason score > or = 7 in men age < or = 70 years with a PSA level < or = 10 ng/ mL (P < 0.05). Markoff models reviewed in the literature demonstrated significant sensitivity to progression, complication, and comorbidity rates. Recent models suggested significant increases in quality adjusted life expectancy for men choosing radical prostatectomy over watchful waiting if they were age < 70 years and had no severe comorbidities. Original cost estimates from benefit-cost analysis showed similar results of cost per carcinoma and cost per quality-adjusted life-year extension as later cost effectiveness models. CONCLUSIONS: Current diagnostic trends toward the persistent increased detection of localized prostate carcinoma in younger men, combined with a marked reduction in distant stage disease, suggest significant potential mortality reductions. These trends may have greater implications for AAM, but further research is needed to produce models of mortality reduction from emerging data. PMID- 9351562 TI - Family history facilitates the early diagnosis of prostate carcinoma. AB - BACKGROUND: Although all men age >50 years are at an increased risk for the development of prostate carcinoma, 2 major factors increase this risk: family history and race. This article outlines the influence of family history on the risk of prostate carcinoma and current understanding of factors that increase this risk. METHODS: Published studies investigating the familial and hereditary link to prostate carcinoma are reviewed. The results of an investigation into the mendelian inheritance of prostate carcinoma are discussed as well as the relation between hereditary cancer syndromes such as breast and ovarian carcinoma and prostate carcinoma. RESULTS: A positive family history of prostate carcinoma increases the relative risk of prostate carcinoma in male first-degree relatives approximately twofold. Prostate carcinoma is inherited as an autosomal dominant trait. The relative risk of prostate carcinoma increases with multiple affected relatives. CONCLUSIONS: Hereditary prostate carcinoma is estimated to be associated with 43% of men in whom the diagnosis of prostate carcinoma is made at age <55 years, 34% of men in whom the diagnosis is made at age <70 years, and only 9% of men diagnosed before age 85 years. Hereditary prostate carcinoma should be suspected in families with an early age at onset of the disease and/or multiple affected family members. Because hereditary prostate carcinoma is characterized by an early age at onset, first-degree relatives in high risk families should begin screening before age 50 years. PMID- 9351563 TI - Results of hospital cancer registry surveys by the American College of Surgeons: outcomes of prostate cancer treatment by radical prostatectomy. AB - BACKGROUND: The number of prostate cancer patients treated by radical prostatectomy has increased. Different data sources have yielded various estimates of the outcomes of this treatment and the need for additional therapy. To provide additional perspective on these issues, the American College of Surgeons conducted surveys of cancer registries and reviewed related data. METHODS: In 1993, in the first phase of the study, hospital cancer registries and programs were sent survey forms and instructions requesting data on up to 5 patients treated by radical prostatectomy at their institutions in 1990. In 1996, in the second phase of the study, additional data were requested on treatment administered to the 1990 patients up to 5 years after surgery, and hospitals were also invited to submit new data on patients diagnosed in 1993. Responses were received from 482 hospitals concerning 2122 patients for 1990, and 265 hospitals provided data on 1304 patients diagnosed in 1993. Follow-up data on 1076 of the 1990 patients were provided by 258 hospitals. Kaplan-Meier survival curves were calculated to determine the probability of additional treatment after radical prostatectomy. RESULTS: Similar surgical pathology outcomes were reported for the 1990 and 1993 patients. For 1990 and 1993, respectively, it was reported that 27.5% and 29.7% of patients maintained erectile function adequate for intercourse after surgery. For 1990 and 1993, respectively, complete control or only occasional urinary incontinence requiring no pads was reported for 81.3% and 79.8% of patients. The surgical mortality rates were less than 1% for both the 1990 and the 1993 patients. The 5-year cumulative probability of any additional treatment after radical prostatectomy was 10.5%. Seminal vesicle involvement, positive surgical margins, lymph node involvement, capsular penetration, high Gleason score, and high prostate specific antigen were significantly associated with greater probability of additional treatment. CONCLUSIONS: Hospital cancer registries are valuable sources of data on patterns of care and outcome for prostate cancer patients. Continuing evaluation of the outcomes of prostate cancer treatments is needed to reconcile the differences in outcomes reported from different data sources. PMID- 9351564 TI - Bizarre leiomyomas of the uterus: a comprehensive pathologic study of 24 cases with long-term follow-up. AB - A series of 24 bizarre (symplastic, pleomorphic) leiomyomas was analyzed to determine their spectrum of gross and microscopic features and to establish their clinical behavior by obtaining long-term follow-up. Patients' ages ranged from 25 to 51 years (mean 40.7). Maximum dimension of the tumors was about 1-14 cm (mean 4.2), with 83% measuring <5.5 cm and 8% > or = 10 cm. Grossly, the tumors generally resembled ordinary leiomyomas; one third had yellow or tan areas, two had hemorrhages, and a few had focal softening, cavitation or myxoid change. When evaluable (50%), the tumor border was generally circumscribed. Cellularity was 1+ in 21%, 2+ in 58%, and 3+ in 21%. Two tumors were predominantly of epithelioid cell type. Bizarre giant cells were unifocal in 12.5%, multifocal in 37.5%, and diffusely distributed in 50%. More than one third of the tumor contained giant cells in 67%. Degenerative changes occurred in 38%, usually hydropic change. One had infarct-type necrosis. None had tumor cell necrosis. Fibrinoid change of blood vessels occurred in 21%, usually accompanied by chronic inflammation. Mitosis counts ranged from zero to 2.8 mitotic figures (MFs)/10 high-power fields (HPFs) (mean 0.8) by the average count method. By the highest count method, the range was 0-7 MFs/10 HPFs (mean 1.6), with 13% having none and 29% having > or = 2 MFs/10 HPFs. Some pyknotic and karyorrhectic nuclei resembled abnormal MFs. Treatment was hysterectomy in 20 and myomectomy in six (including two with subsequent hysterectomy). Complete follow-up was available in 100%, ranging from 1 to 18.9 years (mean 11.2). Postoperative intervals were > or = 10 years in 58% and > or = 5 years in 83% of cases. All patients were alive and well. Although three patients subsequently developed unrelated second primary malignant neoplasms, none developed recurrence or metastasis of bizarre leiomyoma. Bizarre leiomyomas have a wider range of morphologic changes and mitotic activity than previously documented. This study firmly establishes the benign behavior of bizarre leiomyomas, even for those tumors with high cellularity, numerous widely distributed bizarre cells, and mitosis counts in the 2-7 MFs/10 HPFs range by the highest count method. PMID- 9351565 TI - Large cell calcifying Sertoli cell tumor of the testis: contrasting features of six malignant and six benign tumors and a review of the literature. AB - We report six malignant and six benign large cell calcifying Sertoli cell tumors of the testis and compare the features of malignant and benign cases based on these cases and those in the literature. All the tumors in this report consisted of sheets, nests, solid tubules, and cords of eosinophilic cells, with focal calcifications, as well as a substantial neutrophilic infiltrate in 11 of them. Analysis of our cases and those in the literature showed that the malignant tumors were unilateral and solitary and occurred at a mean age of 39 years (range 28-51 years), whereas the benign neoplasms were bilateral and multifocal in 28% of cases and occurred at a mean age of 17 years (range 2-38 years). Only one malignant tumor occurred in a patient with evidence of a genetic syndrome (Carney syndrome), whereas 36% of benign tumors had various genetic syndromes or endocrine abnormalities. Most of the tumors in the latter cases were bilateral and multifocal. There were strong associations of malignant behavior with size >4 cm, extratesticular growth, gross or microscopic necrosis, high-grade cytologic atypia, vascular space invasion, and mitotic rate greater than three mitoses per 10 high-power fields. All malignant cases exhibited at least two of these features, whereas all benign cases lacked any of them. The presence of any one of these features in a solitary large cell calcifying Sertoli cell tumor, especially in a patient >25 years of age, should be viewed as suspicious for malignant behavior, whereas the presence of two or more of these features indicates a strong probability of a malignant course. "Low" percentages (< or =35%) of tumor cells staining for proliferating cell nuclear antigen (PCNA) also may correlate with benign behavior, but some benign tumors have high PCNA values. Ki-67 values (MIB-1 antibody) did not correlate with biologic behavior, nor did immunostains for p53 protein. PMID- 9351566 TI - Primary leiomyosarcoma of bone: a clinicopathologic, immunohistochemical, and ultrastructural study of 33 patients and a literature review. AB - Leiomyosarcoma of bone is a rare tumor in an unusual location. Previous analysis of this entity mostly involved small numbers of cases with limited follow-up. Thirty-three patients with leiomyosarcoma of bone between 1977 and 1996 were studied, and the histologic appearance and grade were correlated with subsequent treatment and clinical behavior. To be included in this study the tumor had to be intraosseous, with other primary sites of origin clinically excluded. Also, most of the sarcomatous tissue (> or =70%) had to be of intramedullary location with only limited extraosseous extension. The patient's age at diagnosis ranged from 13 to 77 years (average 44.4). The gender distribution was equal. The long bones were preferentially affected (64%), with the lower extremity, around the knee joint, predominantly involved. Five patients (15%) developed postradiation leiomyosarcomas. The histologic analysis showed that the osseous leiomyosarcomas are most commonly of the classic type, followed by the epithelioid, myxoid, and pleomorphic variants. Immunoreactivity for smooth muscle markers (smooth muscle actin, common muscle actin, desmin) was positive in all tumors, and ultrastructural confirmation was obtained in 21% of cases. All sarcomas were histologically graded, which accurately reflected the subsequent prognosis. Seventy-five percent of the lesions were high-grade and the rest low-grade. The histologic grade of the tumors correlated with both the recurrence as well as the metastatic rates and together with the clinicopathologic stage of disease represented the cornerstone on which prudent therapy should be based. PMID- 9351567 TI - Angiomyoid and follicular dendritic cell proliferative lesions in Castleman's disease of hyaline-vascular type: a study of 10 cases. AB - Castleman's disease of hyaline-vascular type (HV CD) may rarely be associated with a confusing variety of stromal cell overgrowths and neoplasms. We report here on the pathologic and clinical findings of 10 such cases. In addition to the usual complex histoimmunophenotype of the stroma of HV CD and some unusual features that mimicked neoplasms, we observed focal proliferations of angiomyoid (five cases) and follicular dendritic cell type (five cases). The former were nonneoplastic growths featuring compact tangles of spindle cells, exhibiting immunoreactivity for smooth muscle actin and interpreted as vessel-related pericytes and myoid cells. The latter were neoplastic growths of oval to spindle cells intermixed with lymphocytes; the tumor cells grew in long, intersecting bundles, featured various degree of atypia, and expressed the markers of follicular dendritic cells (CD21, CD35, KiM4p). The two types were clinically distinct. Four of five patients with angiomyoid proliferations were young women, who presented with an abdominal mass and were cured by surgery; that is, they had a clinical profile similar to that of patients with the stroma-rich variant of HV CD. The follicular dendritic cell proliferations were in older patients of either gender presenting with masses at various sites, recapitulating the profile of follicular dendritic cell tumors arising independently from HV CD; in three patients with long-term follow-up, recurrences or metastases developed at various intervals from the initial diagnosis (1 1/6, 3 1/2, and 11 years), and one patient died as a result. This study confirms the potential for, and the variety of, stromal cell proliferations in HV CD. Because their biologic behavior differs, correct identification of these various proliferative lesions is clinically important. PMID- 9351568 TI - Primary cutaneous marginal zone B-cell lymphoma: a recently described entity of low-grade malignant cutaneous B-cell lymphoma. AB - Recently a new classification of primary cutaneous B-cell lymphomas (PCBCLs) has been proposed by the European Organization for Research and Treatment of Cancer (EORTC)--Cutaneous Lymphoma Project Group. The marginal zone B-cell lymphomas (MZLs) were not included as a distinct entity because of insufficient experience and controversial opinions. We have studied 32 patients (M:F ratio 1.5:1; age range 25-93 years; mean age 49.6 years; median age 50 years) to determine the diagnostic criteria of primary cutaneous MZL and the relationship with other low grade malignant PCBCLs. For comparison, three patients with immunocytoma were included in the study. Clinically, patients presented with solitary or clustered reddish or red-brown papules, nodules, and plaques, sometimes surrounded by an erythematous halo. Histopathologic sections showed nodular or diffuse infiltrates involving the dermis and subcutaneous fat. Cytomorphologically small to medium sized cells with indented nuclei and abundant pale cytoplasm (marginal zone cells, centrocyte-like cells) predominated. In addition, scattered blasts, lymphoplasmacytoid cells, and plasma cells were observed below the epidermis and at the periphery of the infiltrates. Reactive germinal centers were present in 75% of the cases. The three cases of immunocytoma showed a more monomorphous pattern with predominance of lymphoplasmacytoid cells. The marginal zone cells showed a CD20+, CD79a+, CD5- and Bcl-2+ immunophenotype. They expressed immunoglobulin G in the majority of the cases. Staining with the monocytoid B cell-related antibody KiM1p gave positive results in all specimens with a typical intracytoplasmic granular pattern. A monoclonal distribution of immunoglobulin light chains was observed in marginal zone cells in 75% of the cases. Germinal centers, when present, were either polyclonal or negative for both kappa and lambda light chains. Monoclonal rearrangement of the JH gene was detected via polymerase chain reaction (PCR) in 18 of 26 investigated specimens. Analysis in 12 patients of the bcl-2/immunoglobulin heavy chain gene rearrangement using PCR yielded negative results. Lesions were treated by surgical excision followed in some patients by local radiotherapy. Systemic antibiotic therapy was administered to three patients, with good response in two. The prognosis is excellent. After a mean follow-up of 47.9 months (range 6-252; median 24) all patients are alive without signs of systemic lymphoma. Primary cutaneous MZL represents a distinct clinicopathologic subtype of low-grade malignant PCBCL. PMID- 9351569 TI - Thymoma with pseudosarcomatous stroma: report of an unusual histologic variant of thymic epithelial neoplasm that may simulate carcinosarcoma. AB - Six cases are described of an unusual type of primary thymic epithelial neoplasm characterized by a biphasic epithelial/spindle cell morphology that closely resembled a carcinosarcoma. The patients were two women and four men 28-70 years of age. The tumors presented clinically as asymptomatic anterior mediastinal masses found incidentally on routine chest radiographs. All patients were treated by complete surgical excision. Grossly, the tumors consisted of well circumscribed, encapsulated masses that measured 6-14 cm in greatest diameter and showed a gray-white, homogeneous, rubbery cut surface. Histologically, the lesions were composed of anastomosing islands and cords of oval to polygonal epithelial cells displaying large nuclei with occasional prominent nucleoli and rare mitotic figures, separated by areas containing a highly cellular spindle cell proliferation without nuclear atypia. Thymic remnants could be identified in the periphery of the lesions in four cases. Immunohistochemical stains showed diffuse strong positivity for keratin and focally for epithelial membrane antigen (EMA) in the epithelial cell component, and strong positivity for vimentin and focally for actin in the spindle cell stromal component. Stains for keratin, EMA, desmin, S-100 protein, and CD34 were negative in the spindle stromal cells in all cases except one, in which EMA positivity was present; CD5 stains were negative in the epithelial cells in all cases examined. Electron microscopic examination in one case showed well-formed desmosomes and tonofilaments in the epithelial elements, as well as features indicative of fibroblastic differentiation in the spindle stromal cells. Because of the unusually florid spindle cell stromal component and the focally atypical features of the epithelial cells, some of these tumors initially were misinterpreted as examples of carcinosarcoma. Clinical follow-up in five cases showed that the patients were alive and without evidence of disease over a period of 5-20 years (mean follow-up 10 years), suggesting a benign or very low grade malignant biologic behavior. The present cases appear to represent an unusual, previously undescribed morphologic variant of thymoma characterized by a prominent pseudosarcomatous stromal component. Because of the distinctive histologic appearance and indolent clinical behavior, these lesions should be distinguished from other more aggressive anterior mediastinal neoplasms displaying a biphasic morphology. PMID- 9351570 TI - Malignant lymphoma of the bladder: evidence from 36 cases that low-grade lymphoma of the MALT-type is the most common primary bladder lymphoma. AB - Patients with malignant lymphoma of the bladder were studied, and three clinical groups were defined: those with primary lymphoma localized in the bladder, lymphoma presenting in the bladder as the first sign of disseminated disease (nonlocalized lymphoma), and recurrent bladder involvement by lymphoma in patients with a history of malignant lymphoma (secondary lymphoma). The differences in these groups regarding lymphoma type, clinical presentation, and clinical outcome were studied. Mayo Clinic Tissue Registry records from 1940 to 1996 were searched to identify patients with lymphomas involving the bladder. The lymphomas were classified based on review of the histology and immunophenotype performed by immunoperoxidase methods. Clinical records were reviewed. Presenting symptoms included urinary frequency, dysuria, hematuria, and lower abdominal and back pain. Primary lymphoma was present in six patients. All were B-cell lineage low-grade lymphomas of the mucosa-associated lymphoid tissue (MALT) type. No patient had recurrent lymphoma or died of lymphoma. Nonlocalized bladder lymphoma occurred in 17 patients; one with low-grade lymphoma of the MALT type, four with follicle center lymphomas, and 12 with large cell lymphomas. Excluding two patients who died postoperatively, median survival was 9 years. Six patients died of lymphoma in the follow-up period. Secondary bladder lymphoma occurred in 13 patients: two with low-grade lymphoma of the MALT type, one with follicle center lymphoma, one with mantle cell lymphoma, and nine with diffuse large cell lymphomas. Median survival in this group was 0.6 years. Low-grade lymphoma of the MALT type was the most frequent type of primary bladder lymphoma and was associated with an excellent prognosis. The bladder can be the presenting site of lymphomatous involvement in patients with more widespread disease. Survival in this group is quite favorable and is presumably dependent on lymphoma histologic type, stage of disease, and other prognostic factors. Bladder involvement by recurrent lymphoma is a sign of widely disseminated disease and is associated with a very poor prognosis. PMID- 9351571 TI - Localized hyperplastic gastropathy of the mucous cell- and mixed cell-type (localized Menetrier's disease): a report of 11 patients. AB - Clinical and pathologic findings in five women and six men with the rare localized form of hyperplastic gastropathy of the mucous cell-(foveolar) or mixed cell-(mucous cell and glandular) type are reported. Upper abdominal discomfort, loss of appetite, loss of weight, and anemia were the principal symptoms. Preoperative hypoproteinemia was documented in two patients. Gross findings consisted of a circumscribed area of giant folds, well demarcated from the surrounding normal-appearing mucosa, located predominantly in the corpus in six patients and predominantly in the antrum in four patients. Histologically they corresponded to an increase in the epithelial cell mass principally of mucous cells with elongated and sometimes cystically dilated foveolae, accompanied by a mild inflammatory infiltrate. This so-called localized form of hyperplastic gastropathy has been known since the first description of the disease but has gained relatively little attention in the literature. However, its recognition seems diagnostically important and pathogenetically interesting. Etiology, pathogenesis, and the natural history are mostly unknown. Five of the 11 patients had concomitant adenocarcinoma of the stomach. In four of them the carcinoma was not located within but outside the area of hyperplasia. Because of that and because of a rather unusual accumulation of other tumors of the gastrointestinal tract in these patients, it is suggested that localized hyperplastic gastropathy could be an indicator of an increased risk for gastrointestinal tumors in general more than a possibly premalignant lesion by itself. PMID- 9351572 TI - Crohn's-like complications in patients with ulcerative colitis after total proctocolectomy and ileal pouch-anal anastomosis. AB - Total proctocolectomy with ileal pouch-anal anastomosis (IPAA) has become an established surgical procedure for ulcerative colitis. Occasional patients who have undergone IPAA develop persistent or recurrent episodes of pouchitis (chronic pouchitis), from which a subset also develop gastrointestinal and systemic complications that are identical to those seen in Crohn's disease. These complications include enteric stenoses or fistulas in the pouch or pouch inlet segment, perianal fistulas or abscesses, pouch fistulas, arthritis, iridocyclitis, and pyoderma gangrenosum. The development of Crohn's-like gastrointestinal complications in a patient with chronic pouchitis frequently engenders concern that the pathologist misinterpreted the proctocolectomy specimen as ulcerative colitis instead of Crohn's disease. We describe eight patients who developed chronic pouchitis and Crohn's-like complications after IPAA and total proctocolectomy. In each case, concern was voiced about misinterpretation of the proctocolectomy specimen as ulcerative colitis instead of Crohn's disease after the development of the Crohn's-like complications. Preoperatively, all eight patients had characteristic clinical, radiographic, and pathologic features of ulcerative colitis. Review of the pathology specimens indicated that all eight had ulcerative colitis. Crohn's-like complications are most likely related to chronic pouchitis, which probably is a form of recrudescent ulcerative colitis within the novel environment of the pouch. A diagnosis of Crohn's disease after IPAA surgery should only be made when reexamination of the original proctocolectomy specimen shows typical pathologic features of Crohn's disease, Crohn's disease arises in parts of the gastrointestinal tract distant from the pouch, pouch biopsies contain active enteritis with granulomas, or excised pouches show the characteristic features of Crohn's disease, including granulomas. There were no histologic differences in the total colectomy specimens between the eight ulcerative colitis study patients and 16 control ulcerative colitis patients who had a favorable clinical outcome after IPAA surgery groups. Crohn's-like complications and chronic pouchitis does not necessarily imply an incorrect original interpretation of ulcerative colitis by the pathologist. PMID- 9351573 TI - The immunophenotypic spectrum of meningeal hemangiopericytoma: a comparison with fibrous meningioma and solitary fibrous tumor of meninges. AB - Despite controversy regarding its histogenesis, meningeal hemangiopericytoma (HPC) is a well-defined clinicopathologic entity exhibiting high rates of recurrence and late extracranial metastasis. It must be distinguished from several benign neoplasms, particularly fibrous meningioma (FM) and solitary fibrous tumor (SFT). To determine the immunoprofile of HPC, we studied 27 meningeal examples, including 13 low-grade and 14 high-grade tumors. For comparison, 20 FMs and eight SFTs of the meninges were also evaluated. The immunotype of HPC included vimentin (85%), factor XIIIa (78%) in individual scattered cells, Leu-7 (70%), and CD34 (33%) in a weak, patchy pattern. Focal desmin and cytokeratin positivity was only occasionally encountered (20% each). The SFT shared a similar immunophenotype, except that CD34 expression (100%) was characteristically strong and diffuse. The FM characteristically expressed epithelial membrane antibody (EMA) (80%) and S-100 protein (80%); CD34 reactivity (60%) was patchy and weak. Both within and among all three tumor types, MIB-1 labeling indices varied widely. Specifically, they were unrelated to tumor grade in HPC. Significant reactivity for p53 protein was detected in 52% of HPCs, 17% of SFTs, and 5% of FMs. Meningeal HPC exhibits a distinct antigenic profile, one enabling the exclusion of other entities in nearly all cases. The rare expression of desmin or cytokeratin in HPC suggests either the occurrence of divergent differentiation or, less likely, the possibility that its distinctive morphology is but a phenotype shared by several types of meningeal sarcoma. PMID- 9351574 TI - Cutaneous involvement in polyvinylpyrrolidone storage disease: a clinicopathologic study of five patients, including two patients with severe anemia. AB - Polyvinylpyrrolidone (PVP), formerly a plasma expander, has continued to be inappropriately used in Taiwan for intravenous injection as a "blood tonic." Five cases of PVP storage disease with cutaneous involvement were studied. Two patients presented with cutaneous eruptions mimicking collagen vascular disease and chronic pigmented purpuric dermatosis. Two other cases were found incidentally: one was with a metastatic tumor and the other in a pemphigus lesion. The fifth case was seen in a blind skin biopsy specimen taken to exclude Niemann-Pick disease after hematologic examination of a bone marrow smear. The latter patient and the patient with a collagen vascularlike disease also had severe anemia and serious orthopedic and neurologic complications due to massive infiltration of PVP-containing cells in the bone marrow with destruction of the bone. Severe irreversible anemia due to PVP storage disease has not been reported before. Three patients admitted having a history of receiving intravenous injection of PVP. The samples obtained from two of them indeed contained 5% PVP as determined by chemical analysis. PVP storage disease can be diagnosed by its histopathologic features. The skin biopsy specimens all showed a variable number of characteristic blue-gray vacuolated cells around blood vessels and adnexal structures with positive tinctorial reactions to mucicarmine, colloidal iron, and alkaline Congo red and negative to periodic acid-Schiff (PAS) and alcian blue. The PVP storage cells were shown to be CD68+ macrophages. The presence of PVP in the skin induced little or no inflammatory reaction. Only the pelvic mass in one patient had a foreign body granuloma formation. Our study showed that systemic parenteral administration of PVP preparation could result in the accumulation of PVP storage cells in the skin, with or without clinical eruptions. The diagnosis of systemic PVP storage disease can be established by performing a skin biopsy for pathologic study. It is important for pathologists and clinicians to be aware of this iatrogenic storage disease to avoid misdiagnosis for hereditary storage disease, osteomyelitis, or signet-ring cell carcinoma. Serious hematologic and orthopedic complications can be caused by repeated massive intravenous injection of PVP. Therefore, PVP preparations should be strictly prohibited for systemic administration. PMID- 9351575 TI - Are primary cutaneous immunocytoma and marginal zone lymphoma the same disease? PMID- 9351576 TI - Florid angiogenesis in mucosa surrounding an ileal carcinoid tumor expressing transforming growth factor-alpha. AB - Carcinoid tumors of the gastrointestinal tract are known to be associated with fibrosis and vascular elastosis, either within the tumor or at distant sites. The current report describes prominent vascular proliferation in the villi extending 38 cm proximal and 15 cm distal to an ileal carcinoid tumor. These villi were expanded by vessels, producing a segmental carpet of multiple small polypoid protrusions around the tumor. Immunohistochemical analysis suggested that the major stromal components were of endothelial and myofibroblastic cell origin. The stroma of the tumor itself had minimal fibrosis and vascularity. To our knowledge, this is the first description of vascular proliferation in the vicinity but distinct from a carcinoid tumor. The demonstration of transforming growth factor-alpha (TGF-alpha) synthesis by tumor cells supports the possibility of a field effect by angiogenic factor(s) secreted by the tumor. PMID- 9351577 TI - Adenomatoid tumor of the heart: report of a case. AB - We report a case of an adenomatoid tumor involving the heart. The lesion was found incidentally at the time of cardiac surgery, measured 1.0 cm, and was poorly demarcated from the adjacent myocardium. Microscopically, the tumor consisted of aggregates of relatively large, epithelioid cells that coalesced to form tubular spaces and occasionally branched into anastomosing channels. The neoplastic cells were strongly immunoreactive with antibodies against cytokeratin. The pathologic features of this unusual cardiac tumor are diagnostic of an adenomatoid tumor, a relatively rare benign neoplasm of mesothelial origin usually found in association with the genital tract. Although rare cases of adenomatoid tumors found outside of the genital tract have been described, including two recently reported pleural tumors, it has not been described to involve the heart. PMID- 9351578 TI - Cytogenetic abnormalities and clonal evolution in an adult hepatoblastoma. AB - Hepatoblastomas usually occur in children < 3 years of age, and only occasional adult cases have been described. To date, 20 cytogenetically abnormal childhood hepatoblastomas have been reported. Karyotypic investigations have shown that most hepatoblastomas are diploid or hyperdiploid, often displaying trisomies for chromosomes 2 and 20. We have cytogenetically investigated an adult hepatoblastoma for which no previous karyotypic data exist. A hypertriploid stemline with multiple numerical and structural chromosomal aberrations, including +2 and +20, was found. In addition, the tumor displayed extensive clonal evolution with 11 subclones. Although the tumor thus displayed some chromosomal abnormalities commonly observed in childhood tumors, providing further support for the importance of these abnormalities in the development of hepatoblastoma, the level of genomic complexity seen in the present case has never been described in childhood hepatoblastomas and may suggest a different etiology or pathogenesis. PMID- 9351579 TI - Lymph node revealing solution: a new method for detection of minute axillary lymph nodes in breast cancer specimens. AB - The staging and prognosis of patients with breast cancer is related to the presence or absence of axillary lymph node involvement. However, in some cases no lymph nodes or too small a number of lymph nodes are revealed by the traditional method of palpating and sectioning the axillary fat. In the present study we demonstrate the usefulness of the lymph node revealing solution (LNRS) in breast cancer. Specimens from 13 patients, in whom <10 lymph nodes were identified in the axilla by the traditional method, were included in the study. After excising the lymph nodes by the traditional method, axillary tissue was immersed in LNRS for 6-12 hours. Additional lymph nodes, which stood out as white chalky nodules, were excised and processed as usual. The LNRS increased the mean number of nodes per case from 6.0+/-2.5 found by the traditional method to 12.54+/-4.61 nodes per case (p < 0.01). The size of the nodes identified by the LNRS was significantly smaller (p < 0.01) than those detected by the traditional method. The LNRS changed the lymph node stage of the disease in four of the studied cases (30%). LNRS seems to be the technique of choice for detection of axillary lymph nodes in cases where the number of detected lymph nodes by the traditional method is too small for accurate staging. PMID- 9351580 TI - Prostate cancer volume. PMID- 9351581 TI - Atypical fibroxanthoma with osteoclastlike giant cells. PMID- 9351582 TI - Genetics of asthma: conference summary. PMID- 9351583 TI - Genetic analysis of a quantitative trait in a mouse model of polycystic kidney disease. AB - The development of a variety of powerful tools for genome analysis has facilitated the ability to genetically map loci which contribute to the variation of a quantitative trait. However, the fact that these traits are often determined as a result of complex genetic interactions has made their analysis considerably more difficult then the molecular characterization of qualitative traits that are monogenic in origin. We have described the use of a novel method of chromosomal exclusion to map the recessive mutation juvenile cystic kidney (jck) to mouse chromosome 11 using an intercross between (C57BL/6J x DBA/2J) F1 jck/+ mice. The severity of polycystic kidney disease (PKD) in the intercross progeny, which could be quantitated as a function of kidney size, was significantly more variable than that found in the parental C57BL/6J strain, suggesting that a modifier locus or loci introduced from DBA/2J affects expression of jck. Two regions (one from DBA/2J on chromosome 10 and a second from C57BL/6J on chromosome 1) were found to be associated with inheritance of a more severe PKD phenotype. The finding of a highly significant association of inheritance of a C57BL/6J-related locus with disease severity was unexpected since the PKD phenotype in this inbred background is mild. This result suggests that inheritance in the affected F2 mice of loci from the two different parental backgrounds results in the more severe phenotype, presumably as a consequence of a direct or indirect interaction between their protein products. This type of effect, which is an example of genetic epistasis, will make the molecular characterization of loci that contribute to complex traits markedly more difficult than the analysis of monogenic disorders. PMID- 9351584 TI - Cytokine manipulation in animal models of asthma. AB - We have developed in C57 Black 6 mice an in vivo model of allergic airway inflammation characterized by the presence of IgE antibodies to an inhaled antigen, peribronchial infiltrates with an increased number of eosinophils, and an increased airway responsiveness to nonantigenic bronchoconstrictor stimuli. In this animal model we have investigated the role of different cytokines in the development of IgE antibodies to inhaled antigen, eosinophilic airway inflammation, and airway hyperresponsiveness. The studies were performed by using knockout mice or by exogenous administration of cytokines or cytokine antagonists. Interleukin-4 (IL-4) knockout mice were unable to develop an allergic eosinophilic airway infiltration and did not produce specific IgE antibodies. Chronic aerosol exposure to antigen also did not induce an increase in airway responsiveness. In studies of wild-type mice, pretreatment with the combination of anti-IL5 and anti-IL-5 receptor antibodies, given in an attempt to fully inhibit the effect of endogenously released IL-5, caused a pronounced inhibition of the antigen-induced airway eosinophilia but did not prevent the increase in airway responsiveness. Treatment with IL-12 during the active immunization prevented airway eosinophilia, production of specific IgE antibodies, and the antigen-induced increase in airway responsiveness. In contrast, administration of IL-12 to actively immunized mice during the aerosol exposure abolished airway eosinophilia and airway hyperresponsiveness without affecting the production of specific IgE. PMID- 9351585 TI - Genetics of native airway responsiveness in mice. AB - The inbred mouse represents a powerful tool for dissecting both simple and complex traits. Genetic studies in the mouse should identify disease genes acting in the same biochemical pathway as in the human. Problems associated with genetic heterogeneity, inability to control environmental conditions, lack of an abundant supply of genetic markers, and ethical considerations regarding human genetic crosses are but some of the reasons to study airway responsiveness in the mouse. At present, only a handful of studies have shed light on the genetics of airway responsiveness; even fewer have sought to identify genetic loci that regulate this trait. It is clear that both genetic and environmental factors influence the asthma phenotype and that genetic background is an important consideration when interpreting segregation analysis data. The controversy over the specific mode of inheritance and number and location of quantitative trait loci (QTL) illustrates the need for additional studies. However, given that numerous candidate loci implicated in the pathogenesis of asthma map near QTLs identified in two recent studies, and given the considerable homology between the human and mouse genome, a targeted search for susceptibility genes is warranted in the human. Ideally, these regions will demonstrate linkage in humans. Thus, further work remains to be done to create detailed maps of the regions of linkage in the mouse, and to ultimately identify gene(s) that modify airway responsiveness. mice. PMID- 9351586 TI - The genetics of allergen-induced airway hyperresponsiveness in mice. AB - Airway hyperresponsiveness (AHR) is a fundamental aspect of asthma that has been shown to be influenced by both environmental and genetic factors. Antigen sensitization and challenge of the A/J inbred mouse strain induced AHR, eosinophilic airway inflammation, and lung goblet cell hyperplasia. We discuss the evidence that supports the role of T helper cells and their subsets in determining the airway inflammatory and contractile responses to antigen in a mouse model. Airway hyperresponsiveness and pulmonary eosinophilic inflammation induced by antigen challenge are associated with a Th2 pattern of cytokine expression in the murine lung. CD4+ T cells mediate the airway reaction to antigen, as depletion of CD4+ T cells attenuates the response. The presence of interleukin (IL)-4 induces the Th2 type of immune response, and this cytokine is required for mice to manifest AHR and inflammation to antigen. The Th1 type of immune response is stimulated by IL-12. Antigen-mediated AHR and inflammation are inhibited by IL-12 administration. Airway hyperresponsiveness in the noninflammatory state (without antigen treatment) is inherited in A/J and C3H/HeJ inbred mouse strains. One quantitative trait locus for AHR in progeny derived from these strains is located on murine chromosome 6. We propose that antigen inducd AHR and inflammation also have heritable components. Based on the available immunological data, genes that influence the balance between Th1 and Th2 cells are logical candidate genes for antigen-induced AHR and inflammation. Knowledge of the genes that determine this phenotype will help us understand the mechanisms of human asthma. PMID- 9351587 TI - Modifications of experimental bronchopulmonary hyperresponsiveness. AB - Bronchopulmonary hyperresponsiveness (BHR) is a hallmark of asthma and other inflammatory diseases of the airways. Animal models of BHR are available in which systemic or local immunizations, followed by acute allergenic provocations into the airways, augment responses to intravenous or intratracheal nonspecific bronchoconstrictor agents. Guinea-pig models are easy to manipulate but have serious handicaps: lack of proper genetics, lack of biomolecular tools, and frequent excess of eosinophils in the bronchoalveolar lavage fluid (BALF). Murine models have proper genetics and molecular tools, and they have the further advantage of being widely used for the study of other pathologies. In many of these studies, interleukin (IL)-5 appears as a major cytokine, produced by Th2 lymphocytes. Interleukin-5 promotes eosinophil differentiation and maturation, recruitment to the airways, and possibly activation. The presence of eosinophils in the airways and in the BALF may be necessary but is not sufficient to support BHR, since intense eosinophilia may be present in its absence. Bronchopulmonary hyperresponsiveness is also induced by the administration of lipopolysaccharide (LPS); in that case, eosinophils are not involved, and the role of neutrophils and of tumor necrosis factor-alpha, even though likely, has not been proven. Comparison of BHR induced by allergen (Th2- and largely eosinophil-dependent) and by LPS (probably macrophage-dependent) should allow for a better understanding of the mechanisms of BHR and for the development of important remedies. PMID- 9351588 TI - Genetics of complex disease: approaches, problems, and solutions. AB - Complex or multifactorial diseases are defined as diseases that are ultimately determined by a number of genetic and environmental factors. Although there are many technologies and strategies that can be used to detect genetic factors influencing complex diseases, these technologies and strategies have inherent limitations. In fact, the very name "complex disease" suggests that the results from relevant studies will not be simple to decipher. Ultimately, both the detection and precise characterization of a factor's contribution to a complex disease are difficult undertakings, because the effect of any one factor may be obscured or confounded by other factors. However, the genetic dissection of complex diseases can be greatly facilitated by paying heed to two very basic distinctions. The first distinction is between complexity at the level of individuals and complexity at the level of populations. The second distinction is between the two sequentially pursued components of gene discovery paradigms: gene identification and gene effect characterization. Although genetic epidemiology, as a research field, is oriented to both components of gene discovery for complex diseases, it is suited to gene effect characterization at the population level more than anything else. This paper reviews the origins of the genetic basis of complex traits, as well as the problems plaguing genetic epidemiologic analysis strategies, with the hope of showing how greater attention to these distinctions, as well as a greater integration of relevant knowledge, can alleviate confusion and shape future investigations. In addition, a new discipline, "phenomics" or "phenometrics," could be initiated that would complement genomic research as presently performed. PMID- 9351589 TI - Linkage and candidate gene studies in asthma. PMID- 9351590 TI - Evidence for multiple genetic susceptibility loci for asthma. AB - Genetic susceptibility to asthma is due to multiple genes that interact with each other and the environment. There are many known environmental influences, such as viral and other respiratory infections and exposure to allergens, air pollutants, and active or passive cigarette smoke (1). Genome-wide screens for asthma and atopy have been completed and show statistical evidence for linkage in different racial groups and population samples (4, 5). Some of these linkages have already been replicated in different studies, and most of them are in chromosomal regions containing relevant candidate genes that may regulate inflammatory processes including cytokine synthesis, T-cell responses, or other immune functions. These associations support the relevance of this genetic approach in understanding susceptibility to and expression of asthmatic and allergic phenotypes. Once specific sequence variants are identified, it will become important to test for gene-environment interaction in order to understand the significance and relative effect of each gene on the overall phenotype. PMID- 9351591 TI - Complexities of the genetics of asthma. AB - The task of deciphering the genetics of asthma is very complex. Recent studies of the familiar segregation of asthma showed that no single gene accounts for a major part of the expression of the disease, and that a polygenic model with some evidence of an oligogenic influence (i.e., a handful of loci being responsible for most of the genetic control) provided the best fit to the data. Although a final common pathway of recurrent bronchial obstruction is present in most cases of asthma, the disease shows marked phenotypic variability, suggesting etiologic heterogeneity and strong environmental influences. In an effort to circumvent these obstacles, linkage studies for genes controlling for apparently simpler phenotypes have been attempted. Total serum immunoglobulin E (IgE) levels, for example, show strong familiar aggregation and are known to be strongly correlated with asthma risk. Recent epidemiologic studies have suggested, however, that the inherited component of total serum IgE may be of little relevance as a determinant of asthma. Sensitization to certain aeroallergens is also associated with increased prevalence of asthma and is likely to have a genetic component, but the aeroallergens involved vary markedly with locale. In addition, sensitization to aeroallergens occurring at an early age is more strongly associated with asthma risk than late allergic sensitization, suggesting genetic heterogeneity. Therefore, studies of the genetics of phenotypes known to be strongly associated with asthma may clarify the causal role (if any) of the genes regulating their expression in the pathogenesis of asthma. PMID- 9351592 TI - Epidemiological study of the genetics and environment of asthma, bronchial hyperresponsiveness, and atopy: phenotype issues. AB - The Epidemiological Study of the Genetics and Environment of Asthma (EGEA) combined a case-control study and a family study. The total sample of 1,854 consisted of 348 patients with asthma selected through chest clinics and 416 control subjects and nuclear families ascertained through the cases. The protocol included standardized questionnaires, bronchial responsiveness, allergen skin prick tests according to international protocols, total serum immunoglobulin E (IgE) level measurements, and blood eosinophilia. Criteria used to select subjects with asthma and determine asthma status of relatives for affected sibling pair linkage analysis are described. Based on figures from the 348 asthma cases of the EGEA study, issues relative to the definition of severe asthma and intermediate phenotypes such as bronchial responsiveness and allergic markers are discussed. Given the phenotypic heterogeneity involved, relevant phenotypes that may lead to the detection of genetic factors will depend on the hypothesis tested. Standardization of primary data and subphenotypes is a prerequisite for pooling data, which will be needed in the future to better understand the genetics and environmental factors of asthma. PMID- 9351593 TI - Genetics of asthma: the University of Toronto Program. University of Toronto Genetics of Asthma Research Group. AB - In 1991, we initiated a study to identify susceptibility gene(s) predisposing individuals to the development of asthma. Our strategy was to focus on the collection of inbred populations because (1) they are likely to exhibit greater homogeneity than outbred populations, potentially important in multigenic diseases, (2) there may be fewer genes explaining the asthma diathesis in families with a "founder effect," (3) it is possible to follow the pattern of inheritance more closely, and (4) environmental factors are likely to be more uniform in geographically isolated poulations. We have identified, and in some cases collected data on inbred and/or isolated populations in Brazil, China, Easter Island, Israel, and Tristan da Cunha. We have completed a genome-wide scan on samples from Tristan da Cunha based on a coverage of approximately 20 cM between markers. In addition, we have collected hundreds of outbred individuals from Canada; these data have been helpful in narrowing a linked region based on the transmission/disequilibrium test. PMID- 9351594 TI - Approaches toward the genetic analysis of complex traits: asthma and atopy. AB - With the challenges emerging from the analysis and interpretation of the human genome, and the specific issues pertinent to pursuing the Genome Project itself, it is truly an exciting time in the development of the biological sciences. The occasion is certainly ripe for the emergence of new concepts and ideas, as the theories of complexity, natural selection, and reductionism become integrated into a new whole. We need to learn how to approach the analysis of the complex data sets that will be generated by the Genome Project and address, more generally, the problems inherent in the analysis of the complex diseases such as asthma. Finally, we need to consider how the recent advances in genetics and genomics will affect biomedical research into the next millennium and beyond. PMID- 9351595 TI - Human leukocyte antigen associations in occupational asthma induced by isocyanates. AB - Exposure to diisocyanates is recognized as a leading cause of occupational asthma. Occupational asthma induced by isocyanates shares many characteristics with immunoglobulin E (IgE)-mediated asthma: in both, the responsible agent is known, and the clinical presentation, response to inhalation challenge in the laboratory, and response to antiasthma drugs are similar. Although asthma mediated by an IgE mechanism occurs in atopic subjects, occupational asthma induced by isocyanates occurs mostly in nonatopic asthmatics, and an IgE-mediated mechanism has not been consistently demonstrated. However, activated T lymphocytes, methacromatic cells, and eosinophils are increased in the bronchial mucosa of allergic and nonallergic asthmatics and subjects with occupational asthma induced by isocyanates, suggesting similar, probably immunologically mediated mechanisms for both nonoccupational and occupational asthma. Occupational asthma occurs in up to 5-10% of the exposed subjects. Evaluation of major histocompatibility complex (MHC) class II genes in exposed subjects who develop toluene diisocyanate (TDI) asthma has shown a negative association with HLA-DQB1*0501 and a positive association with HLA-DQB1*0503 alleles. In addition, a high proportion of TDI asthmatics express the HLA-DQB1*0503-associated aspartic acid at residue 57, suggesting that HLA-DQ may have a key role in conferring susceptibility. Thus, asthma induced by the low-molecular-weight agent TDI may result from an immunologic reaction due to the interaction of genetic susceptibility with exposure in the workplace. PMID- 9351596 TI - The candidate region approach to the genetics of asthma and allergy. AB - To date, the strongest linkage claims for genes underlying asthma and atopy have been for chromosomes 5 and 11. Chromosome 5q contains the cytokine cluster and the beta2 adrenoceptor, and the beta chain of the high-affinity IgE receptor (FCepsilonRI-beta) is encoded on chromosome 11q13. We have attempted to replicate these findings in two distinct sample populations from the United Kingdom. Allelic associations were identified in both regions, but there was no significant evidence for linkage. Although we could not substantiate the existence of the nucleotide changes reported within exon 6 of the FCepsilonRI beta gene, an amino acid substitution in exon 7 was strongly linked to asthma and atopy. We have also identified positive linkage and allelic associations to several markers on chromosome 12q in both our UK populations. Independent evidence from another study also supports linkage to 12q, so although our data could not confirm linkage to chromosomes 5 or 11, we have identified an additional region of the genome that could be important for the genetic predisposition to asthma and atopy. PMID- 9351597 TI - Genetics of atopy and asthma: the rationale behind promoter-based candidate gene studies (IL-4 and IL-10). AB - The genetics of atopy and asthma has become a very interesting area for research. Potential candidate genes identified either by the immunopathogenesis of asthma or bronchial hyperresponsiveness, or uncovered by the whole-genome screen, will lead to new and better ways of diagnosing asthma and, more importantly, the potential for drug discovery related to the products of the candidate genes identified in the various genome screening efforts. The candidate gene approach has been applied to the promoter region of a number of cytokine genes, both within and outside of the human 5q33 cytokine gene cluster. As a prototype for both cytokines, work relating to an interleukin (IL)-4 promoter polymorphism and an IL-10 promoter polymorphism will be reviewed as providing a potential molecular mechanism for dysregulation of these cytokine genes in asthma. PMID- 9351598 TI - Polymorphisms of the beta2-adrenergic receptor and asthma. AB - Several missense mutations (polymorphisms) within the coding block of the beta adrenergic receptor (beta2AR) gene on chromosome 5q31 have been identified in the human population. In studies utilizing site-directed mutagenesis and recombinant expression, three loci at amino acid positions 16, 27, and 164 have been found to significantly alter receptor function. The Ile164 form displays altered coupling to adenylyl cyclase, the Gly16 receptor displays enhanced agonist-promoted downregulation, and the Glu27 form is resistant to downregulation. The frequencies of these various forms of the beta2AR are not different in asthmatics than in normal populations. However, given the importance of beta2AR in modulating lung function, studies have been carried out to determine if polymorphic forms may play roles in promoting asthmatic phenotypes, establishing bronchial hyperreactivity, or influencing the response to acute or chronic beta agonist therapy. The results of case-control and family studies to date support these notions. Thus beta2AR polymorphisms act as disease modifiers in asthma and represent one of probably many genetic variables involved in the pathophysiology of asthma. PMID- 9351599 TI - Is "crop rotation" of antibiotics the solution to a "resistant" problem in the ICU? PMID- 9351600 TI - Patient-assessed health outcomes in chronic lung disease: what are they, how do they help us, and where do we go from here? PMID- 9351601 TI - Scheduled change of antibiotic classes: a strategy to decrease the incidence of ventilator-associated pneumonia. AB - The purpose of this study was to determine the impact of a scheduled change of antibiotic classes, used for the empiric treatment of suspected gram-negative bacterial infections, on the incidence of ventilator-associated pneumonia and nosocomial bacteremia. Six hundred eighty patients undergoing cardiac surgery were evaluated. During a 6-mo period (i.e., the before-period), our traditional practice of prescribing a third generation cephalosporin (ceftazidime) for the empiric treatment of suspected gram-negative bacterial infections was continued. This was followed by a 6-mo period (i.e., the after-period) during which a quinolone (ciprofloxacin) was used in place of the third-generation cephalosporin. The incidence of ventilator-associated pneumonia was significantly decreased in the after-period (n = 327) compared with the before-period (n = 353) (6.7 versus 11.6%; p = 0.028). This was primarily due to a significant reduction in the incidence of ventilator-associated pneumonia attributed to antibiotic resistant gram-negative bacteria (0.9 versus 4.0%; p = 0.013). Similarly, we observed a lower incidence of bacteremia attributed to antibiotic-resistant gram negative bacteria in the after-period compared with the before-period (0.3 versus 1.7%; p = 0.125). These data suggest that a scheduled change of antibiotic classes can reduce the incidence of ventilator-associated pneumonia attributed to antibiotic-resistant gram-negative bacteria. PMID- 9351603 TI - Exogenous surfactant and partial liquid ventilation: physiologic and pathologic effects. AB - We compared the effects of surfactant and partial liquid ventilation (PLV), and the impact of administration order, on oxygenation, respiratory system compliance (Crs), hemodynamics, and lung pathology in an animal lung injury model. We studied four groups: surfactant alone (S; n = 8); partial liquid ventilation alone (PLV-only; n = 8); surfactant followed by partial liquid ventilation (S PLV; n = 8); and partial liquid ventilation-followed by surfactant (PLV-S; n = 8). Following treatments, all animals had improved oxygenation index (OI) and Crs. Animals in PLV groups showed continued improvement over 2 h (% change OI: PLV-S -83% versus S -47%, p < 0.05; % change Crs: S-PLV 73% versus S 13%, p < 0.05). We also saw administration-order effects: surfactant before PLV improved Crs (0.92 ml/cm H2O after surfactant versus 1.13 ml/cm H2O after PLV, p < 0.02) without changing OI, whereas surfactant after PLV did not change Crs and OI increased (5.01 after PLV versus 8.92 after surfactant, p < 0.03). Hemodynamics were not different between groups. Pathologic analysis demonstrated decreased lung injury in dependent lobes of all PLV-treated animals, and in all lobes of S PLV animals, when compared with the lobes of the S animals (p < 0.05). We conclude that surfactant therapy in combination with PLV improved oxygenation, respiratory system mechanics, and lung pathology to a greater degree than surfactant therapy alone. Administration order affected initial physiologic response and ultimate pathology: surfactant given before PLV produced the greatest improvements in pathologic outcomes. PMID- 9351602 TI - Primary importance of zwitterionic over anionic phospholipids in the surface active function of calf lung surfactant extract. AB - The relative contributions of zwitterionic and anionic phospholipids to the surface-active function of calf lung surfactant extract (CLSE) were assessed by measurements of surface properties in vitro and pressure-volume (P-V) mechanics in excised rat lungs in situ. Surface activity and mechanical effects were compared for chromatographically purified CLSE subfractions containing the complete mix of phospholipids (PPL) or modified phospholipids depleted in anionic components (mPPL), alone or combined with 1.3% (by weight) of hydrophobic surfactant proteins (SP-B and SP-C). Surface pressure-time (pi-t) adsorption isotherms at 37 degrees C were very similar for dispersions of PPL and mPPL in a Teflon dish with a stirred subphase to minimize diffusion resistance. Combination of either PPL or mPPL with hydrophobic SP substantially improved adsorption, but mixtures of PPL:SP and mPPL:SP had only small differences in pi-t isotherms and reached the same final equilibrium pi of approximately 47 mN/m achieved by CLSE. Surface pressure-area (pi-A) isotherms and maximum surface pressures were also very similar for spread films of PPL versus mPPL and PPL:SP versus mPPL:SP on the Wilhelmy balance (23 degrees C and 37 degrees C). Respreading based on pi-A isotherm area calculations was slightly better in surface-excess films of PPL versus mPPL and PPL:SP versus mPPL:SP, but differences were minor and were smaller at 37 degrees C than at 23 degrees C. Overall dynamic surface activity in oscillating bubble studies was not significantly different for PPL versus mPPL or for PPL:SP versus mPPL:SP, and the latter two mixtures both reached minimum surface tensions < 1 mN/m (37 degrees C, 20 cycles/min, 0.5 mM phospholipid). Dispersions of PPL:SP, mPPL:SP, and CLSE were also not significantly different in improving P-V mechanics almost to normal when instilled in lavaged, excised rat lungs at 37 degrees C (30 mg/2.5 ml saline). These data suggest that zwitterionic phospholipids have a major role over anionic phospholipids in interacting with hydrophobic SP in the adsorption, dynamic surface tension lowering, film respreading, and pulmonary mechanical activity of the hydrophobic components of calf lung surfactant in CLSE. PMID- 9351604 TI - Diaspirin crosslinked hemoglobin improves systemic oxygen uptake in oxygen supply dependent septic rats. AB - Diaspirin crosslinked hemoglobin (DCLHb) is a cell-free hemoglobin derived from human erythrocytes. DCLHb has been shown to improve blood flow to vital organs in healthy and septic animals. In this study, we determined the efficacy of DCLHb by comparing its effect on systemic O2 uptake to freshly stored and aged red blood cells (RBCs) in septic rats. Twenty-four hours after induction of sepsis by cecal ligation and perforation, O2 supply dependency was created by isovolemic hemodilution with rat plasma. In O2 supply dependency, rats were randomized to receive an exchange transfusion of 7.5 ml "fresh" RBCs (stored < 6 d; Hct: 70%), "fresh" diluted RBCs (stored < 6 d; Hct: 30%), "old" RBCs (stored 28 to 35 d; Hct: 70%), or DCLHb (Hb: 100 g/L). We found, that survival following O2 supply dependency and transfusion with old RBCs was poor (33% versus 91.7% in the other groups; p < 0.01), precluding further analysis of post-transfusion data from this group. Systemic O2 uptake increased in all remaining groups (p < 0.001), while systemic O2 delivery increased with "fresh" RBCs (p < 0.0001) and "fresh" diluted RBCs (p < 0.05) but not with DCLHb. Systemic O2 extraction increased with DCLHb as compared to baseline (p < 0.05) and to the other groups (p < 0.0001). Improved tissue oxygenation was associated with an increase in blood pressure and a fall in arterial lactate in all groups. We conclude that transfusion of DCLHb or "fresh" RBCs was efficacious at increasing systemic O2 uptake in O2 supply dependent, septic rats. PMID- 9351605 TI - Reduced lipid peroxidation and ischemia-reperfusion injury after lung transplantation using low-potassium dextran solution for lung preservation. AB - Ischemia-reperfusion injury is one of the significant problems in clinical lung transplantation. We investigated the effect of lung preservation with Euro Collins solution (EC group) or low-potassium dextran solution (LPD group) on lipid peroxidation and ischemia-reperfusion injury in a pig model of lung allotransplantation. The donor lungs were preserved at 4 degrees C for 18 h. Left sided single lung transplantation was performed, followed by 6 h of reperfusion. Lipid peroxidation was measured as thiobarbituric acid-reactive materials (TBARM) in bronchoalveolar lavage (BAL) fluid and effluent solutions from pulmonary artery (Effluent). After 18 h of ischemia, the LPD group showed lower TBARM in BAL and Effluent than the EC group (p < 0.05). After ischemia plus reperfusion, lung wetto-dry weight ratios and TBARM levels in BAL in the LPD group were lower than those of the EC group (p < 0.05). Lung wet-to-dry weight ratios correlated with TBARM levels in BAL (p < 0.05, r = 0.50). We conclude lipid peroxidation in BAL and Effluent may reflect the degree of ischemia-reperfusion injury, and lung preservation with LPD can reduce lipid peroxidation and lung injury as compared with EC. PMID- 9351606 TI - Impairment of lung and chest wall mechanics in patients with acute respiratory distress syndrome: role of abdominal distension. AB - Recent data have suggested that the elastic properties of the chest wall (CW) may be compromised in patients with ARDS because of abdominal distension (4). We partitioned CW and lung (L) mechanics, assessed the role of abdominal distension, and verified whether the underlying disease responsible for ARDS affects the impairment of respiratory mechanics. Volume-pressure (V-P) curves (interrupter technique) were assessed in nine patients with surgical ARDS and nine patients with medical ARDS. Relative to nine patients undergoing heart surgery, V-P curves of the respiratory system (rs) and L of patients with surgical or medical ARDS showed a rightward displacement. V-P curves of the CW and the L showed an upward concavity in patients with medical ARDS and a downward concavity in patients with surgical ARDS. Although the CW and the abdomen (abd) V-P curves in patients with medical ARDS were similar to those obtained in patients undergoing heart surgery, they showed a rightward shift and a downward flattening in patients with surgical ARDS. In five of these patients, a reduction in static end-inspiratory pressure of the abd (69+/-4%), rs (30+/-3%), CW (41+/-2%), and L (27+/-3%) was observed after abdominal decompression for acute bleeding. Abdominal decompression therefore caused an upward and leftward shift of the V-P curves of the respiratory system, chest wall, lung, and abdomen. In conclusion we showed that impairment of the elastic properties of the respiratory system may vary with the underlying disease responsible for ARDS. The flattening of the V-P curve at high pressures observed in some patients with ARDS may be due to an increase in chest wall elastance related to abdominal distension. These observations have implications for the assessment and ventilatory management of patients with ARDS. PMID- 9351607 TI - Lower respiratory tract colonization and infection during severe acute respiratory distress syndrome: incidence and diagnosis. AB - Ventilator-associated pneumonia (VAP) is difficult to detect and is often unsuspected during adult respiratory distress syndrome (ARDS). We prospectively evaluated lower respiratory tract (LRT) colonization and infection in 30 patients with severe ARDS (PaO2/FIO2 ratio < 150 mm Hg), using repeated quantitative cultures of plugged telescopic catheter (PTC) specimens taken blindly via the endotracheal tube every 48 to 72 h after onset of ARDS. All patients except one were receiving antibiotics. When VAP was suspected on the presence of clinical criteria for infection, a repeated PTC and, when possible, a bronchoalveolar lavage (BAL) were obtained before any new antimicrobials were administered; samples growing > or = 10(3) cfu/ml (PTC) or > or = 10(4) cfu/ml (BAL) were considered diagnostic of infection. Twenty-four VAP episodes were diagnosed in 18 patients (60% of patients or 4.2/100 ventilator-days) a mean of 9.8+/-5.7 d after onset of ARDS. Eighteen LRT colonization episodes were recorded; 16 of 24 (66%) VAP episodes were preceded (by 2 to 6 d) by LRT colonization with the same organism(s), and only two episodes of colonization were not followed by VAP. We conclude that although VAP is of relatively late-onset during severe ARDS, its incidence is much higher than in other conditions and can be underestimated. Lower airways colonization is consistently followed by infection with the same organisms and precedes VAP in two thirds of episodes. Repeated protected specimens taken blindly may provide a useful means to predict infection and therefore allow early antimicrobial therapy in high-risk patients with diffuse lung injury. PMID- 9351608 TI - Lactate production by the lungs in acute lung injury. AB - Arteriovenous differences in lactate (AVLAC) across the lungs are usually small and close to zero. However, it has recently been reported that the lungs can produce increased amounts of lactate in some patients with acute respiratory distress syndrome (ARDS). The aim of this study was to evaluate lactate production in various types of acute lung injury requiring mechanical ventilation and hemodynamic monitoring. Since the differences involved are usually small, minor errors in lactate measurement could greatly influence AVLAC. Based on an analysis of these errors (see text for details), we averaged five arterial and venous samples for each measurement. We investigated 122 patients: 43 with acute lung injury (ALI), nine with cardiogenic pulmonary edema (CPE), 37 with bronchopneumonia (BPN), seven with single lung transplantation (LTX), and 26 with other causes of respiratory failure (OTHER). There was no difference in arterial lactate between the various groups. AVLAC was higher in patients with ALI than in the other groups (0.20+/-0.23 versus 0.07+/-0.11 mEq/L). In patients with ALI, AVLAC was proportional to the Murray's lung injury score (-0.032+/-0.032x; r = 0.46, p < 0.01). Lung lactate production was calculated as the product of the cardiac index times AVLAC and was significantly higher in patients with ALI than in the other groups (0.69+/-0.88 versus 0.19+/-0.30 mEq/min; p < 0.05). In patients with ALI, lung lactate production was inversely related to the PaO2/FIO2 (1.42 - 0.005x; r = 0.35, p < 0.05) but directly related to the venous admixture (-0.36 + 0.003x; r = 0.49, p < 0.01) and the lung injury score (-0.19 + 0.36x; r = 0.45, p < 0.01). Lung lactate production was not significantly related to arterial lactate levels. These data indicate that AVLAC and lung lactate production can be increased in patients with ARDS but remain within the normal range in other types of respiratory failure. PMID- 9351609 TI - The systemic inflammatory response in the development of ventilator-associated pneumonia. AB - Ventilator-associated pneumonia (VAP) is the most frequent occurring infection among mechanically ventilated patients. The clinical presentation of VAP ranges from relatively benign to a severe illness with septic shock. The influence of VAP on patient outcome has not been elucidated and its effects on the inflammatory response of the host are unknown. In a case-control study, the systemic inflammatory response was investigated in patients developing VAP as compared with control patients matched on duration of mechanical ventilation and underlying diseases. Patients developing VAP (n = 42) were matched to a single control (without VAP), who was matched on seven variables. VAP was diagnosed with bronchoscopic techniques. The inflammatory response, reflected by circulating levels of interleukin-6 (IL-6) and interleukin-8 (IL-8), was determined on the day of diagnosis (or day of matching for controls), 4 and 2 d before diagnosis, and 2 d after diagnosis. The development of VAP was not associated with an increase in circulating levels of IL-6 or IL-8. Among patients in which VAP was associated with a clinical presentation of severe sepsis or septic shock (n = 10), IL-6 and IL-8 levels increased and were higher than in the corresponding controls. Moreover, 60% of cases with severe sepsis or septic shock died as compared with 20% of their matched controls (p = 0.06). Mortality rates were similar in patients with uncomplicated VAP and their matched controls (25% and 34%, respectively). High circulating levels of IL-6 and IL-8 were associated with higher mortality rates. The clinical picture of VAP can be subdivided into different types, ranging from uncomplicated to an infection associated with severe sepsis or septic shock, elevated circulating levels of IL-6 and IL-8, and an increased mortality rate. PMID- 9351610 TI - Normoxic lung ischemia/reperfusion accelerates shedding of angiotensin converting enzyme from the pulmonary endothelium. AB - Normoxic lung ischemia/reperfusion (I/R) leads to oxidative injury of the pulmonary tissue. We analyzed angiotensin-converting enzyme (ACE) in perfused rat lungs upon I/R in order to assess the endothelial injury produced. I/R led to a time-dependent increase in ACE activity in the perfusate, from 145+/-14 mU to 252+/-1 mU, and to reduction of ACE activity in the lung tissue homogenate, from 29.7+/-2.3 U to 22.7+/-1.7 U. About 80% of ACE activity in control and I/R rat lungs was associated with an aqueous phase of extracted perfusates, thus indicating that I/R accelerates shedding of the hydrophilic form of ACE from the plasma membrane. To specifically assess ACE localized on the luminal surface of the pulmonary endothelium, we perfused rat lungs with a radiolabeled monoclonal antibody (mAb) to ACE (anti-ACE mAb 9B9). Pulmonary uptake of mAb 9B9 with I/R was reduced from 32.1+/-1.7% to 24.8+/-0.9%. In contrast, I/R led to a marked increase in the pulmonary uptake of nonspecific [125I]IgG, from 0.17+/-0.02% to 0.67+/-0.04%. Lung wet weight was equal to 0.78+/-0.08% of body weight in the I/R group versus 0.57+/-0.02% at the control level. The observed increase in [125I]IgG uptake and wet lung weight indicate that I/R causes an increase in lung vascular permeability. These results indicate that normoxic lung I/R induces injury to the pulmonary vascular endothelium. PMID- 9351611 TI - Health-related quality of life after acute lung injury. AB - Our study objective was to assess health-related quality of life in survivors of acute lung injury (ALI) and to supplement generic and disease-specific questionnaires with findings from a focus group of ALI survivors. Six patients participated in the focus group, which revealed patient concerns with amnesia, depressed mood, avoidance behaviors, and a prolonged recovery period. Using a cross-sectional study design, 24 patients completed a questionnaire 6 to 41 mo after their lung injury. A total of 43% of the patients with ALI met criteria for depression; 43% had self-reported significant functional limitations, although 39% had minimal or no limitations. Significant respiratory and psychologic symptoms were reported in a quarter to a third of patients. There were large decrements in all domains of the SF-36 (a generic health-related quality-of-life instrument) in our sample compared with norms previously established for the general population. In addition, our patients had similar physical difficulties compared with previously studied patients with chronic medical illnesses but had more deficits in the social functioning and mental health domains. We conclude that long after lung injury, survivors have significantly lower health-related quality of life than the general population and are likely to have pulmonary and psychologic symptoms. PMID- 9351612 TI - Effect of hyperoxic hypercapnia on variational activity of breathing. AB - Dysrhythmias of breathing occur in several clinical disorders, but their mechanistic basis is obscure. To understand their pathophysiology, factors responsible for the variability of breathing need to be defined. We studied the effect of hyperoxic hypercapnia (CO2) on the variational activity of breathing in 14 volunteers before and after delivering CO2 nonobstrusively via a plastic hood. Compared with air, CO2 increased the gross variability of minute ventilation (VI) and tidal volume (VT), and decreased that of inspiratory time (TI) and expiratory time (TE) (all p < 0.03). CO2 increased the autocorrelation coefficient at a lag of one breath for VI (p < 0.05), the number of consecutive breath lags having significant autocorrelation coefficients for VI and VT (both p < 0.01), and the cycle time of oscillations in VI (p = 0.03) and VT (p = 0.04). Uncorrelated random behavior constituted > or = 80% of the variance of each breath component, correlated behavior represented 9 to 20%, and oscillatory behavior represented < 1% during both air and CO2. CO2 increased the correlated behavior of volume components, which was accompanied by development of low-frequency oscillations with a cycle time consistent with central chemoreceptor activation. PMID- 9351613 TI - Increased carbon monoxide in exhaled air of asthmatic patients. AB - Exhaled carbon monoxide (CO) concentrations were measured on a CO monitor by vital capacity maneuvers in asthmatic patients receiving or not receiving inhaled corticosteroids and in nonsmoking and smoking healthy control subjects. CO was detectable and measured reproducibly in the exhaled air of all subjects. The exhaled CO concentrations were higher in asthmatic patients not receiving inhaled corticosteroids (5.6+/-0.6 ppm, p < 0.001) and similar in asthmatic patients receiving inhaled corticosteroids (1.7+/-0.1 ppm) compared with those in nonsmoking healthy control subjects (1.5+/-0.1 ppm). Smoking healthy control subjects had the highest levels of exhaled CO concentration among the groups (21.6+/-2.8 ppm, p < 0.001). To examine whether inhaling corticosteroids reduce exhaled CO concentration in a given asthmatic patient, 12 patients with symptomatic asthma who were being treated by inhaled beta2-agonists alone underwent measurements of exhaled CO concentration before and 4 wk after the initiation of inhaled corticosteroid treatment. All patients had reductions in exhaled CO concentration (p < 0.001) and eosinophil cell counts in sputum (p < 0.01) that were accompanied by an improvement in airway obstruction. Changes in exhaled CO concentration were significantly related to those in the eosinophil cell counts in sputum (p < 0.001). The present study shows an elevation of exhaled CO in asthmatic patients that decreases with corticosteroid therapy. Increases in the exhaled CO levels therefore may reflect inflammation in the asthmatic lung. PMID- 9351614 TI - Treatment of nocturnal airway obstruction improves daytime cognitive performance in asthmatics. AB - It has been shown that asthmatics have nocturnal symptoms associated with impaired cognitive performance. We explored more carefully different therapeutic approaches on this performance in relation to lung function in 46 atopics with mild to moderate asthma and with a circadian variation in peak expiratory flow (PEF) > or = 15%. In a double-blind, parallel study they inhaled salmeterol 50 microg or fluticasone 250 microg or a combination of both twice daily for 6 wk. The psychometric tests used informed about focused attention, mental flexibility, concentration, and attention. The results of the psychometric tests were compared with those in healthy control subjects. The PASAT score and the finishing time of the color-word chart subtest were significantly lower in these asthmatics than in the control subjects. Circadian PEF variation was the only independent factor significantly associated with impaired cognitive performance before the treatment period. The three treatment groups were equally effective in reducing circadian PEF variation below 10% and in improving FEV1 and bronchial hyperresponsiveness to methacholine (MCh) both day and night. After 6 wk of therapy, the daytime cognitive performance was improved to levels comparable to those of the healthy control subjects no matter which drug was inhaled. We conclude that a high level of circadian PEF variation (> or = 20%) has been associated with lower daytime cognitive performance in asthmatics. Reduction of circadian PEF variation to below 10% is an important goal of treatment in asthmatics. PMID- 9351615 TI - Airway responsiveness to sulfur dioxide in an adult population sample. AB - We determined the prevalence of airway hyperresponsiveness to sulfur dioxide (SO2) in an adult population sample of 790 subjects 20 to 44 yr of age. Subjects were drawn randomly from the population of Hamburg, Northern Germany, within the framework of the European Community Respiratory Health Survey. In addition, we analyzed the relationship between SO2 responsiveness and a number of risk factors, such as a history of respiratory symptoms, methacholine responsiveness, and atopy derived from skin-prick test results. SO2 inhalation challenges were performed during isocapnic hyperventilation at constant rate (40 L x min(-1), for 3 min) with doubling concentrations of SO2 up to a maximum concentration of 2.0 ppm. If subjects achieved a 20% decrease in FEV1 from baseline during the challenge, they were considered to be hyperresponsive to SO2. The raw prevalence of SO2 hyperresponsiveness within the population sample studied was 3.4% (95% confidence interval [CI]: 2.3 to 5.0%). Adjustment for nonparticipation led to an estimated prevalence of SO2 hyperresponsiveness of 5.4%. Among subjects with hyperresponsiveness to methacholine, 22.4% (95% CI: 20.1 to 25.3) demonstrated hyperresponsiveness to SO2. There was no significant correlation between the degrees of hyperresponsiveness to methacholine and SO2. Predictors of a positive SO2 response were hyperresponsiveness to methacholine (p < 0.0001), a positive history of respiratory symptoms (p < 0.05), and a positive skin-prick test to at least one common allergen (p < 0.05). We conclude from these data that airway hyperresponsiveness to SO2 can be found in about 20 to 25% of subjects within the 20- to 44-yr age range who are hyperresponsive to methacholine. PMID- 9351616 TI - Dose-response relationship to inhaled endotoxin in normal subjects. AB - Exposure to endotoxin and to its purified derivative lipopolysaccharide (LPS) is related to several occupational pulmonary diseases and to severe domestic asthma. An inhalation of a given dose of pure LPS produces both a systemic and a bronchial inflammatory response. Information on the dose-response relationship to inhaled LPS in normal subjects is a prerequisite to define the safety threshold of exposure. In the present study, the clinical and inflammatory responses to rising doses of inhaled LPS was evaluated. Nine normal volunteers were challenged weekly by inhalation with saline, 0.5, 5, and 50 microg LPS (Escherichia coli). The response determinators are the clinical symptoms, fever, FEV1, blood polymorphonuclear neutrophils (PMNs) with their level of activation (measured by luminol enhanced-chemiluminescence), and both the blood and the urine concentrations of the C-reactive protein (CRP). To assess the bronchial inflammatory response, an induced sputum was obtained 6 h after each dose of LPS, and the total and differential cell counts as well as the MPO, ECP, and TNF-alpha concentrations were measured. Compared with the saline, an inhalation of 0.5 microg LPS induces a significant decrease in the PMN luminol-enhanced chemiluminescence (p < 0.01), which could reflect a process of margination and/or extravascular sequestration of activated PMN. Inhalation of 5 microg LPS is associated with a significant rise in blood CRP (p < 0.01) and PMNs (p < 0.001) and in sputum PMNs (p < 0.05), monocytes (p < 0.05), and MPO (p < 0.05). Inhalation of 50 microg LPS was characterized by a significant increase in temperature (p < 0.01), blood PMNs (p < 0.001), blood and urine CRP (p < 0.01 and < 0.01), and sputum PMNs (p < 0.001), monocytes (p < 0.05), lymphocytes (p < 0.05), MPO (p < 0.01), TNF-alpha (p < 0.01), and ECP (p < 0.01) while five subjects develop symptoms. In normal subjects, the response to inhaled LPS is dose-related, the most sensitive markers of LPS-induced inflammation being the blood PMNs count with their level of activation, the blood CRP concentration, and the sputum PMNs count. The no-response threshold to an acute inhalation of LPS is less than 0.5 microg. PMID- 9351618 TI - Volume dependence of respiratory impedance in infants. AB - We previously studied low-frequency respiratory impedance (Zrs) data at an elevated lung volume to separate airway and tissue mechanical properties in normal infants (Am. I. Respir. Crit. Care Med. 1996; 154:161-166). The aim of the present study was to determine the volume dependence of the airway and tissue mechanics by extending Zrs measurements to lower lung volumes. Zrs spectra between 0.5 and 21 Hz were measured in supine sleeping infants (n = 8; 7 to 26 mo of age) at mean transrespiratory pressures (Ptr[mean]) of 20, 10, and 0 cm H2O, during periods of apnea induced by inflating the infants' lungs to a pressure of 20 cm H2O through a face mask. At each inflation pressure, a model containing airway resistance (Raw) and inertance (law) and tissue damping (G) and elastance (H) was fitted to Zrs data. At FRC, the values of Raw, law, G, and H were 20.6+/ 4.9 (SD) cm H2O x s/L, 0.037+/-0.014 cm H2O x s2/L, 39.6+/-10.3 cm H2O/L, and 147+/-35 cm H2O/L, respectively. Increase of Ptr(mean) caused a monotonous decrease in Raw (42+/-7% of the value at FRC), while law remained constant. The tissue parameters were minimal at a Ptr(mean) of 10 cm H2O (68+/-10% and 78+/-6% in G and H, respectively) and significantly higher at both 0 and 20 cm H2O. Although Zrs measurements can be made in most infants at lung volumes as low as FRC, an inflation pressure of 20 cm H2O provides a higher success rate and is therefore a more suitable condition for general use. PMID- 9351617 TI - An evaluation of colchicine as an alternative to inhaled corticosteriods in moderate asthma. National Heart, Lung, and Blood Institute's Asthma Clinical Research Network. AB - Colchicine demonstrates an array of anti-inflammatory properties of potential relevance to asthma. However, the efficacy of colchicine as an alternative to inhaled corticosteroid therapy for asthma is unknown. Five centers participated in a controlled trial testing the hypothesis that in patients with moderate asthma needing inhaled corticosteroids for control, colchicine provides therapeutic benefit as measured by maintenance of control when inhaled steroids are discontinued. Subjects were stabilized on triamcinolane acetonide (800 microg daily) and then enrolled in a 2-wk run-in during which all subjects took both colchicine (0.6 mg/twice a day) and triamcinolone. At the end of the run-in, all subjects discontinued triamcinolone and were randomized to continued colchicine (n = 35) or placebo (n = 36) for a 6-wk double-blind treatment period. The treatment groups were similar in terms of disease severity. After corticosteroid withdrawal, 60% of colchicine-treated and 56% of placebo-treated subjects were considered treatment failures as defined by preset criteria. No significant difference in survival curves was found between treatment groups (log rank = 0.38). Other measures, including changes in FEV1, peak expiratory flow, symptoms, rescue albuterol use, and quality of life scores, also did not differ between groups. Of note, subjects failing treatment had significantly greater methacholine responsiveness at baseline than did survivors (PC20, 0.81+/-1.38 versus 2.11+/-2.74 mg/ml; p = 0.01). An analysis of treatment failures suggested that the criteria selected for failure reflected a clinically meaningful but safe level of deterioration. We conclude that colchicine is no better than placebo as an alternative to inhaled corticosteroids in patients with moderate asthma. Additionally, we conclude that the use of treatment failure as the primary outcome variable in an asthma clinical trial where treatment is withdrawn is feasible and safe under carefully monitored conditions. PMID- 9351619 TI - Bronchial lability and responsiveness in school children born very preterm. AB - We evaluated bronchial lability and responsiveness in 29 prematurely born children (birth weight < 1,250 g) 8 to 14 yr of age, 12 with histories of bronchopulmonary dysplasia (BPD). Flow-volume spirometry, a bronchodilator test, and histamine challenge at the office and home monitoring of peak expiratory flow (PEF) values twice daily for 4 wk with and without a beta2-agonist were performed with a novel device, the Vitalograph Data Storage Spirometer. The spirometric values at the office and the results of home monitoring were compared with those for a control group of children born at term. All spirometric values except FEV1/FVC were significantly lower in the BPD group than in the non-BPD group (p < 0.0001). Ten children (83%) in the BPD group and four (24%) in the non-BPD group had subnormal spirometric values at the office, indicating bronchial obstruction. Of the children with obstruction, 79% reported respiratory symptoms during the preceding year, and 57% had increased diurnal PEF variation and/or responded to administration of a beta2-agonist during home monitoring or at the office. The BPD children were significantly more responsive to histamine than the non-BPD children (p = 0.002). All spirometric values were significantly lower in both preterm groups than in the control group born at full term (p < 0.01). In conclusion, regardless of BPD, bronchial obstruction, bronchial lability, and increased bronchial responsiveness are common in prematurely born children of school age. PMID- 9351620 TI - Effect of prone and supine positions on functional residual capacity, oxygenation, and respiratory mechanics in ventilated infants and children. AB - Although numerous reports have described the improvement in PAO2 in patients in the prone position, the underlying mechanism has yet to be determined. Some authors have suggested this phenomenon may be related to an increase in functional residual capacity (FRC); however, no previous studies have described positional changes in FRC in children with severe lung disease or in those under neuromuscular blockade. We measured arterial blood gases, FRC, Rrs, and Crs in supine and prone positions in 30 patients under neuromuscular blockade with lung disorders including moderately severe restrictive (n = 10) and obstructive (n = 10) disease and control subjects without significant lung disease (n = 10). Prone positioning was not associated with a significant increase in FRC in the cohort of 30 patients, nor in any of the subgroups. Although individual patients demonstrated large improvements in oxygenation, a statistically significant (but clinically insignificant) increase in AaPO2 ratio was observed only in the subgroup of patients with obstructive disease (0.35+/-0.03 to 0.38+/-0.04, p = 0.027). There was no correlation between changes in FRC and changes in AaPO2 (r = 0.225, p = 0.23). A significant improvement in Rrs occurred in the prone position compared to supine in patients with obstructive lung disease, decreasing from 0.264+/-0.024 to 0.216+/-0.021 cm H2O/ml/s, p = 0.009. No significant changes in Crs were seen in the prone position. We conclude that prone positioning has no effect on FRC and in this series of 30 patients significantly improved oxygenation only in patients with obstructive airway disease. A significant decrease in Rrs in patients with obstructive lung disease was also observed. PMID- 9351621 TI - Controlled trial of inhaled budesonide in patients with cystic fibrosis and chronic bronchopulmonary Psuedomonas aeruginosa infection. AB - The efficacy and safety of anti-inflammatory treatment with inhaled glucocorticosteroids in patients with cystic fibrosis (CF) and complicating chronic Pseudomonas aeruginosa (P.a.) lung infection was studied in a placebo controlled, parallel, double-blind single center trial. Active treatment consisted of budesonide dry powder, 800 microg twice daily, delivered from a Turbuhaler. The study period covered two successive 3-mo intervals between elective courses of intravenous anti-Pseudomonas antibiotics. Fifty-five patients entered the study, with a mean age of 20 yr and a mean FEV1 of 63% of predicted. Analysis of all patients entered, irrespective of trial adherence ("intention to treat"), showed a decrease in FEV1 in the first period of -0.032 L in patients on budesonide versus -0.187 L in patients on placebo (p = 0.08). The corresponding figures for the patients adhering to the protocol during the first period were 0.017 L versus -0.198 L (p < 0.05, confidence interval of the difference: -0.035 to +0.327 L). For all patients entered, as well as for patients adhering to the trial, there was always a trend in favor of budesonide, as judged by changes in FEV1 and FVC in both 3-mo periods. None of the patients had asthma, but the patients on budesonide had a mean improvement in histamine reactivity of +1.15 dose steps over the entire 6-mo period, as opposed to +0.017 dose steps in patients on placebo (p < 0.05). There was also a significant (p = 0.01) correlation between pre-trial histamine reactivity and the change in FEV1 in the first period in patients on budesonide. We conclude that inhaled glucocorticosteroids can be of short-term benefit in patients with CF and chronic P.a. infection and that those patients most likely to benefit from this treatment are patients with hyperreactive airways. Prolonged studies in larger number of patients are necessary to determine the long-term efficacy of this treatment. PMID- 9351622 TI - Lower airway inflammation in infants and young children with cystic fibrosis. AB - Airway inflammation is an important component of cystic fibrosis (CF) lung disease. To determine whether this begins early in the illness, before the onset of infection, we examined bronchoalveolar lavage (BAL) fluid from 46 newly diagnosed infants with CF under the age of 6 mo identified by a neonatal screening program. These infants were divided into three groups: 10 had not experienced respiratory symptoms or received antibiotics and pathogens were absent in their BAL fluid; 18 had clear evidence of lower respiratory viral or bacterial (> or = 10(5) CFU/ml) infection; and the remaining 18 had either respiratory symptoms, taken antibiotics, or had < 10(5) CFU/ml of respiratory pathogens. Their BAL cytology, interleukin-8, and elastolytic activity were compared with those from 13 control subjects. In a longitudinal study to assess if inflammation develops or persists in the absence of infection, the results of 56 paired annual BAL specimens from 44 CF infants were grouped according to whether they showed absence, development, clearance, or persistence of infection. In newly diagnosed infants with CF, those without infection had BAL profiles comparable with control subjects while those with a lower respiratory infection had evidence of airway inflammation. In older children, the development and persistence of infection was accompanied by increased inflammatory markers, whereas these were decreased in the absence, or with the clearance, of infection. We conclude that airway inflammation follows respiratory infection and, in young children, improves when pathogens are eradicated from the airways. PMID- 9351623 TI - Ventilation-perfusion mismatch in patients with pleural effusion: effects of thoracentesis. AB - Pleural effusion (PE) often causes abnormal pulmonary gas exchange. Thoracentesis is commonly used to relieve dyspnea in patients with PE, but its effect upon arterial oxygenation is varied and poorly understood. This investigation sought to: (1) characterize the distribution of ventilation-perfusion (VA/Q) ratios in patients with PE and (2) assess the effects of PE drainage by thoracentesis upon pulmonary gas exchange. We studied nine patients (two females) with a mean age of 39+/-20 (SD) yr. All of them had PE of recent clinical onset (< 2 wk of symptoms), without other apparent medical conditions. Before thoracentesis, PaO2 was 82.3+/-10.2 mm Hg and AaPO2 was 28.7+/-10.0 mm Hg. Patients had broadened unimodal VA/Q distributions with small amounts of blood flow perfusing lung units with low VA/Q ratios (< 0.1) (1.4+/-2.2%) and mild intrapulmonary shunt (6.9+/ 6.7%). PaO2 was significantly related to the amount of shunt (rho = -0.82; p < 0.01) but not to the percentage of blood flow perfusing low VA/Q units. While thoracentesis drained 693+/-424 ml of fluid and caused a significant fall in mean pleural pressure (by -10.7 +/- 7.1 mm Hg; p < 0.01), PaO2, AaPO2, and shunt remained unchanged; only the amount of blood flow perfusing low VA/Q ratios increased slightly (2.4+/-2.6%; p < 0.05). This study shows that: (1) intrapulmonary shunt is the main mechanism underlying arterial hypoxemia in patients with PE and (2) effective thoracentesis has minor short-term effects upon pulmonary gas exchange. These findings are in accord with delayed (> 30 min) pulmonary volume re-expansion after thoracentesis with or without the coexistence of mild ex vacuo pulmonary edema. PMID- 9351624 TI - End-inspiratory airway occlusion: a method to assess the pressure developed by inspiratory muscles in patients with acute lung injury undergoing pressure support. AB - We evaluated the end-inspiratory occlusion maneuver as a means to estimate the inspiratory effort during pressure support ventilation (PS). In nine nonobstructed acute lung injury (ALI) patients, we applied four levels of PS (0, 5, 10, 15 cm H2O) to modify the inspiratory effort. End inspiratory occlusions (2 to 3 s) were performed at the end of each experimental period by pushing the inspiratory hold button of the ventilator (Servo 900 C; Siemens, Berlin, Germany). We took the difference between the end-inspiratory occlusion plateau pressure and the airway pressure before the occlusion (PEEP + PS) as an estimate of the inspiratory effort and called it PMI (Pmusc,index). From the esophageal pressure tracing we obtained a reference measurement of the pressure developed by the inspiratory muscles at end inspiration (Pmusc,ei) and of the pressure-time product per breath (PTP/b) and per minute (PTP/min). In each patient, PMI was correlated with Pmusc,ei (p < 0.01) and PTP/b (p < 0.01). A PMI threshold of 6 cm H2O detected PTP/min < 125 cm H2O s/min with a sensitivity of 0.89 and a specificity of 0.89. We conclude that PMI is a good estimate of the pressure developed by the inspiratory muscles in ALI patients and may be used to titrate PS level. The major advantage of PMI is that it can be obtained from the ventilator display without any additional equipment. PMID- 9351625 TI - Surfactant proteins-A and -B are elevated in plasma of patients with acute respiratory failure. AB - Surfactant protein-A (SP-A) leaks into the circulation of patients with acute respiratory distress syndrome (ARDS) or acute cardiogenic pulmonary edema (APE) in a manner inversely related to lung function. Since surfactant protein-B (SP-B) is synthesized as a precursor considerably smaller than alveolar SP-A, we investigated whether it enters the circulation more readily. Reactivities consistent with SP-B proprotein (approximately 42 to approximately 45 kD) and the approximately 25 kD processing intermediate were detected in plasma. Plasma immunoreactive SP-B levels were significantly higher in ARDS (8,007+/-1,654 ng/ml [mean+/-SEM], n = 22) and APE (3,646+/-635 ng/ml, n = 10) patients compared with normal subjects (1,685+/-58 ng/ml, n = 33) and ventilated patients with no cardiorespiratory disease (1,829+/-184 ng/ml, n = 7). All groups had plasma SP B/SP-A ratios approximately 6- to approximately 8-fold higher than in normal lavage or ARDS tracheal aspirate fluid, consistent with protein sieving. During admission, both plasma SP-B and the SP-B/SP-A ratio were inversely related to blood oxygenation (PaO2/FIO2) (p < 0.0001 and p < 0.025, n = 260 from 39 patients; Spearman) and static respiratory system compliance (deltaV/deltaP) (p < 0.0001 and p < 0.01, n = 168 from 25 patients). We describe in detail three patients and conclude that immunoreactive SP-B enters more readily than SP-A, is cleared acutely, and provides a better indicator of lung trauma. PMID- 9351626 TI - Interleukin-1 in ischemia-reperfusion acute lung injury. AB - Interleukin-1 (IL-1) is a proinflammatory cytokine produced by blood-borne and resident inflammatory lung tissue involved in the thrombotic occlusion of the pulmonary microcirculation and the increase of the vascular permeability following a wide variety of injuries and sepsis. The locally accentuated, organ related activation of this cytokine seems to be responsible for the development of acute lung injury. The present study was conducted to determine if IL-1beta was produced in an ischemia-reperfusion (I/R) rat model subjected to lung injury. We measured sequential perfusate levels of IL-1beta by ELISA and we measured IL-1 gene expression in the rat lung tissue by a reverse-transcriptase polymerase chain reaction method. Little IL-1beta gene expression was observed in normal rat lung tissue. Perfusate IL-1beta slightly increased 2 h after induced ischemia and 3 h after reperfusion. IL-1beta gene expression rapidly increased as early as 30 min after ischemia and continued to increase for up to 120 min. IL-1beta gene expression was dramatically upregulated during reperfusion after cessation of ischemia, reached a peak at 1 h, and then gradually decreased (2 to 3 h) to near baseline levels. During ischemia, the increased IL-1 gene expression was not significantly different between the ventral and dorsal sites of the lung. However, IL-1 gene expression markedly increased on the dorsal part (the dependent site for a rat in a supine position) after reperfusion. From these results, it appears that IL-1 may have an important role in I/R lung injury. PMID- 9351627 TI - Bacterial pneumonia causes augmented expression of the secretory leukoprotease inhibitor gene in the murine lung. AB - The cDNA of murine secretory leukoprotease inhibitor (SLPI) was cloned from a mouse lung cDNA library. The amino acid sequence deduced from the cDNA showed 58 and 51% homology with those of human and porcine SLPI, respectively. A two-domain structure with similar amino acid sequences, four intradomain disulfide bonds, and high proline content, which are characteristics common to human and porcine SLPI, was also found in the mouse protein. The amino acid residues for the signal sequence and active site are also conserved in mouse SLPI. RNase protection assay showed the expression of the SLPI gene in liver, intestine, spleen, and epididymis, suggesting the distribution of SLPI in tissues other than lung and seminal vesicles. In the lung infected with Streptococcus pneumoniae strain FP1284, 10 h after inoculation of bacteria the number of SLPI mRNA transcripts was three times higher than baseline. The increased level of expression remained constant for at least 48 h. This result clearly contrasts to that obtained for spleen, in which the SLPI mRNA transcript level was mostly unchanged during the course of pneumonia. These facts suggested the local regulation of the SLPI gene expression in vivo in response to inflammatory stimuli at the site of inflammation. PMID- 9351628 TI - Quantitative detection of human cytomegalovirus DNA in lung transplant recipients. AB - Human cytomegalovirus (HCMV) disease remains a major cause of morbidity and mortality after lung transplantation. Currently, routine diagnostic tests for HCMV are inefficient and insensitive or nonspecific for HCMV disease. We describe an efficient, highly sensitive, quantitative polymerase chain reaction (PCR) assay for HCMV using competitive PCR and fluorescently labeled primers, and we have used this to measure HCMV DNA load in donor and recipient tissues of six lung transplant recipients at the time of transplantation, and 2 wk after transplantation when clinically stable. Total DNA yield was adequate for analysis in transbronchial biopsy, bronchoalveolar lavage, and peripheral blood leukocytes, but the endobronchial biopsy specimens did not consistently produce sufficient DNA for analysis. There was a large intersubject and intrasubject variability between tissues in HCMV DNA load, with a tendency for greater levels in lung tissue compared with BAL or peripheral blood cells. All six HCMV IgG seronegative donors or recipients were found to have HCMV DNA present. One of the three seronegative matched transplant recipients developed histopathologically proven HCMV disease, and HCMV DNA levels were shown to increase at that time point and subsequently decrease with ganciclovir treatment. This assay will allow prospective studies to confirm the predictive value of HCMV DNA load in donor and recipient tissues for HCMV disease. PMID- 9351629 TI - Contrasting effects of hypochlorous acid and hydrogen peroxide on endothelial permeability: prevention with cAMP drugs. AB - Activated polymorphonuclear leukocytes generate a cascade of reduced oxygen metabolites. In addition to their antimicrobial role, hydrogen peroxide (H2O2) and hypochlorous acid (HOCl) function as inflammatory mediators and increase the protein permeability of the vascular endothelium. The objectives of the present study were to compare the effects of H2O2 and HOCl with respect to relative potencies and the time course and magnitude of changes in cell shape and permeability of endothelial cell monolayers derived from bovine pulmonary artery, to determine if HOCl produced by conversion of H2O2 with myeloperoxidase and Cl- produces comparable results as the direct administration of HOCl, and to show that adenosine 3',5'-cyclic monophosphate (cAMP)-enhancing agents can prevent the increased endothelial permeability induced by HOCl and H2O2. HOCl given directly or produced by myeloperoxidase, H2O2, and Cl- caused faster and greater changes in cell shape (cell retraction), electrical resistance, and protein permeability (125I-labeled albumin clearance) of endothelial cell monolayers than induced by H2O2. HOCl (10 to 100 microM) induced these changes within 1 to 3 min, whereas H2O2 (50 to 400 microM) required approximately 30 min. 8-Bromo-cAMP prevented the increased endothelial protein permeability induced by HOCl or H2O2, but isoproterenol only prevented the H2O2 response. Thus, HOCl at a much lower concentration caused a faster and greater increase in endothelial permeability in vitro than H2O2, and an increased intracellular level of cAMP prevented the increased permeability induced by either oxidant. PMID- 9351630 TI - Comparison of murine nasal-associated lymphoid tissue and Peyer's patches. AB - The nasal mucosal is the first site of contact with inhaled antigens. However, the nature of local immune responses and the role of nasal-associated lymphoid tissue (NALT) in those responses have rarely been studied. To characterize the cells involved in mucosally derived immune responses, NALT and Peyer's patch (PP) cells from normal mice, and mice immunized intragastrically or intranasally with cholera toxin (CT), were isolated and analyzed. Compared with PP cells, unstimulated NALT cells contained a higher proportion of T-cells. The CD4:CD8 ratio in NALT cell preparations was less than that observed in PP and more closely resembled that seen in spleen. Additionally, the total B-cell frequency in NALT cell isolates was 20% lower than that observed in PP cell preparations. Although NALT and PP cell isolates contained both mature B-cells and cells undergoing activation to express surface IgA, unlike PP, NALT showed no significant frequency of IgA-switched cells. After intranasal immunization with CT, toxin-specific IgA antibody-forming cells (AFCs) were detected in NALT cell preparations. The numbers of these cells correlated with CT-specific IgA in nasal, but not in gut washes or sera, thus suggesting local nasal production of antigen-specific mucosal antibodies. There was no evidence of anti-CT AFCs in NALT or CT-specific antibody in nasal washes after intragastric CT administration. These results support the notion that nasal mucosal antibody production is best achieved via direct stimulation of IgA-committed, NALT-derived B-cells. PMID- 9351631 TI - Ribavirin therapy for adenovirus pneumonia in an AIDS patient. AB - We report the effectiveness of ribavirin in an AIDS patient with multinodular pneumonia due to adenovirus. A 38-year-old AIDS patient who experienced multiple opportunistic infections and whose CD4 lymphocyte count was 5/mm3 developed bilateral nodular lung opacities. Lung surgical biopsy yielded necrotizing pneumonitis with characteristic nuclear inclusions and positive immunocytology with adenovirus antibodies. Marked clinical and radiological improvement was obtained after intravenous then oral ribavirin. Ribavirin was discontinued after 40 d because of anemia. Relapse of pneumonia with respiratory distress led to death 8 mo later. This observation illustrates a rarely reported pulmonary opportunistic infection in AIDS and the potential value of ribavirin therapy for adenovirus pneumonia. PMID- 9351632 TI - Enteral feeding improves outcome and protects against glycerol-induced acute renal failure in the rat. AB - Acute renal failure is a common cause of morbidity and mortality in critically ill patients and frequently results from vasoconstrictive ischemic injury to the kidney. Protein and amino acids can vasodilate renal blood vessels. Thus, we tested the hypothesis that enteral feeding could prevent renal ischemic injury using an experimental model in which renal vasoconstriction is believed to cause ischemic renal injury. This study was performed using male Sprague-Dawley rats, and renal injury was induced by glycerol injection into the hind limbs. The resulting muscle necrosis (rhabdomyolysis) causes acute renal injury. In the first part of the study, 35 animals were randomized to a peptide-based enteral diet or water via a duodenal feeding tube and subsequently injected with glycerol. Seventy-eight percent (14 of 18) of the animals receiving the enteral diet survived 3 d compared with 35% (six of 17) of the water-fed animals (p < 0.05). Blood urea nitrogen (47+/-8 versus 137+/-27 mg/dl) and creatinine (0.8+/ 0.1 versus 2.0+/-0.3 mg/dl) were significantly lower in the enteral survivors than in the water survivors. In the second part of the study, renal plasma flow (para-aminohippurate clearance) and glomerular filtration rate (insulin clearance) were measured in similarly treated animals (n = 14) 1 d after injury. Renal plasma flow (4.83+/-0.65 versus 2.37+/-0.62 ml/min) and glomerular filtration rate (2.05+/-0.27 versus 0.89+/-0.22 ml/min) were significantly higher in the enteral group than in the water group. These data indicate that enteral feeding can prolong survival and decrease renal injury after glycerol-induced rhabdomyolysis. The mechanism for the protection is partly related to maintenance of renal blood flow. PMID- 9351633 TI - Chest radiographic findings in patients with tuberculosis with recent or remote infection. AB - To determine if chest radiographic findings differ in adult tuberculosis patients with recent and remote infection, we reviewed the chest radiographs of 103 patients with tuberculosis in Los Angeles and performed RFLP analyses of their Mycobacterium tuberculosis isolates. Patients whose isolates had identical or closely related RFLP patterns were considered a "cluster." Most patients in large clusters (more than seven patients) had tuberculosis from recent infection, whereas most unclustered patients had tuberculosis from remote infection. Mediastinal adenopathy or pleural effusions were classified as typical of recent infection, and upper lobe infiltrates, cavitation, or fibrosis were classified as characteristic of remote infection. Radiographic patterns were typical of remote infection in 62% of patients and were characteristic of recent infection in 23% of patients. The distribution of these radiographic patterns was similar in clustered and unclustered patients, both with or without human immunodeficiency virus (HIV) coinfection. However, mediastinal adenopathy and pleural effusions were significantly more common in HIV-infected patients. We conclude that: (1) chest radiographic findings in adults with tuberculosis of recent infection are similar to those in patients with remote infection; (2) the distinctive chest radiographic findings in HIV-infected patients with tuberculosis are not due to an increased frequency of recent infection. PMID- 9351634 TI - Dose-response effect for adrenal suppression with repeated twice daily inhaled fluticasone propionate and triamcinolone acetonide in adult asthmatics. AB - A single blind randomized crossover trial was performed comparing placebo (PL); low (L), medium (M) and high (H) doses of fluticasone propionate (FP) L: 330 microg, M: 770 microg, H: 1,540 microg per day and triamcinolone acetonide (TAA) L: 400 microg, M: 800 microg, H: 1,600 microg per day. Each drug was given twice daily over a total of 9 d, with 3 d for each dose level. Each 9-d drug sequence was preceded by a 3-d placebo, and was separated by a 12-d washout period. Twelve mild-to-moderate, stable adult asthmatics, mean (SEM) age, 34.3 (2.9) yr, mean FEV1: 82.1 (2.0) % predicted, and FEF25-75%: 53.6 (5.5) % predicted, receiving up to 400 microg of inhaled corticosteroid per day, were studied. After each 3-d treatment period, blood samples were taken for 8:00 A.M. serum cortisol. Ten-hour overnight urine collections were taken for measurement of urinary cortisol and corrected for creatinine excretion, starting at 10:00 P.M. following the sixth dose. For 8:00 A.M. serum cortisol compared with PL there was significant (p < 0.001) dose-related suppression with FP but not with TAA, which amounted to a 2.03-fold ratio for H FP versus H TAA. For corrected urinary cortisol/creatinine excretion, there was a significant (p < 0.005) dose-related suppression for FP but not for TAA. This amounted to a 1.9-fold ratio for H FP versus H TAA. For doses < 1,000 microg/d, the number of individual results with an abnormal low urinary cortisol value (< 10 nmol/10 h) were: 10/24 for FP versus 3/24 for TAA (p < 0.005). In conclusion, for 8:00 A.M. serum cortisol and overnight corrected urinary cortisol/creatinine excretion, there was significant dose-related suppression with FP but not with TAA. For both of these parameters at the highest dose of both drugs, this amounted to a two-fold ratio in suppression. PMID- 9351635 TI - The effect of deep inspiration on methacholine dose-response curves in normal subjects. AB - Normal subjects develop exaggerated airway narrowing when deep inspiration (DI) is voluntarily suppressed during methacholine challenge. Failure of periodic inflation may interfere with the bronchodilating effect of DI, and this may be fundamental to the difference in bronchodilation caused by DI in asthmatics and normal subjects. To determine whether repeated exhalations to residual volume (RV) and/or incomplete inspiration to baseline total lung capacity (TLC) could contribute to exaggerated narrowing during challenge, we tested 10 subjects on three separate days using modified methacholine challenge protocols. On Day 1, partial and complete flow volume curves were obtained after each dose. On Day 2, DI was prohibited, but partial curves were performed. On Day 3, DI and exhalation to RV were prohibited. TLC was measured pre- and post-challenge on each day. After comparable doses of methacholine, there was a greater change in FEV1 on Day 2 (27+/-15) and Day 3 (38+/-17) than on Day 1 (14+/-8) (p < 0.05). There were no differences in changes in FEV1 and FVC between Days 2 and 3, or in TLC between all 3 d. We conclude that exaggerated airway narrowing occurs in normal subjects when DI is prohibited and that this effect is not due to repeated expiration to RV, nor due to an artifact caused by a failure to inhale to TLC. PMID- 9351636 TI - Fair allocation of intensive care unit resources. American Thoracic Society. PMID- 9351637 TI - Age-related changes in lung structure and function in the senescence-accelerated mouse (SAM): SAM-P/1 as a new model of senile hyperinflation of lung. PMID- 9351638 TI - Cingulate function in depression. PMID- 9351639 TI - A role for the tumour suppressor gene p53 in regulating neuronal apoptosis. AB - The tumour suppressor gene p53 is a nuclear phosphoprotein whose correct functioning is crucial for an appropriate cellular response to DNA damage. It has been suggested that p53 may act as a 'guardian of the genome' since when DNA damage is mild, p53 functions to halt cell cycle progression allowing DNA repair to occur before progression through the cell cycle. This prevents 'fixing' of lesions into the genome during replication. However when DNA damage is severe and irreversible, p53 induces the cell to undergo apoptosis. Recent studies have demonstrated DNA fragmentation and increased expression of p53 within neurons after injury. It appears that p53 expression may precede DNA fragmentation suggesting that rather than being induced in neurons in response to DNA damage, p53 expression may actually initiate neuronal apoptosis leading to DNA fragmentation. Recent reports documenting the resistance of neurons derived from p53-null mice (p53-/-) to excitotoxicity and DNA damaging agents both in vitro and in vivo and showing that p53 overexpression induces neuronal apoptosis in vitro support a role for the tumour suppressor gene p53 in regulating neuronal apoptosis. Here we review the recent evidence and discuss likely mechanisms involved in p53-mediated neuronal apoptosis. PMID- 9351640 TI - Substrate-bound carbohydrates stimulate signal transduction and neurite outgrowth in an olfactory neuron cell line. AB - Subpopulations of olfactory receptor neurons, which are dispersed throughout the olfactory neuroepithelium, express specific cell surface carbohydrates and project to discrete regions of the olfactory bulb. Cell surface carbohydrates such as N-acetyl-lactosamine have been postulated to mediate sorting and selective fasciculation of discrete axon subpopulations during development of the olfactory pathway. Substrate-bound N-acetyl-lactosamine promotes neurite outgrowth by both clonal olfactory receptor neuron cell lines and olfactory receptor neurons in vitro, indicating that cell surface carbohydrates may be ligands for receptor-mediated stimulation of axon growth in vivo. In the present study, the role of transmembrane signaling in N-acetyl-lactosamine-stimulated neurite outgrowth was examined in the clonal olfactory neuron cell line 4.4.2. Substrate-bound N-acetyl-lactosamine stimulated neurite outgrowth which was specifically inhibited by antagonists to N- and L-type calcium channels and to tyrosine kinase phosphorylation. These results indicate that N-acetyl-lactosamine can evoke transmembrane receptor-mediated responses capable of influencing neurite outgrowth. PMID- 9351642 TI - Effects of benzodiazepine receptor agonists in neurones acutely dissociated from the rat neostriatum. AB - To understand the properties of benzodiazepine receptor in the neostriatum, we examined the potentiating effects of diazepam, triazolam and brotizolam on the GABA(A) receptor-mediated Cl- current in dissociated rat neostriatal neurones using the nystatin-perforated patch recording configuration. Neurones were classified into large and small neurones, on their somatic size. In the large neurones, which are putative cholinergic interneurones, all the benzodiazepine receptor agonists recognized a single effective site. However, in the small neurones, which are mostly considered to be projecting neurones, the effect of brotizolam was best described when two effective sites were assumed. Therefore, the properties of benzodiazepine receptor differed among large and small neurones while at least two kinds of functional binding sites were also found to exist in small neurones. PMID- 9351641 TI - Cloning and expression of human 5-HT4S receptors. Effect of receptor density on their coupling to adenylyl cyclase. AB - We have isolated a cDNA encoding the 5-HT4S receptor by RT-PCR on poly (A)+ RNA from both human heart and brain. The sequence homology with the rat and mouse 5 HT4 receptors was high: 93.8% of identity in the amino acid sequence. None of the 24 amino acid substitutions observed between rat and human receptors are at positions likely to modify their pharmacology. Comparing the pharmacological properties of six agonists and five antagonists on rat and human 5-HT4S receptors revealed no significant differences. We have analyzed the behavior of renzapride, a full and a partial agonist on mouse colliculi neurons and human heart biological responses respectively. The coupling efficiency of renzapride was two fold lower than that of 5-HT for the stimulation of 5-HT4S receptors transfected in two different cell lines (LLC-PK1 and COS-7), but increasing the receptor density suppressed the partial agonist effect of renzapride. PMID- 9351643 TI - Activity-dependent conduction velocity changes of A(delta) fibers in a rat model of neuropathy. AB - Activity-dependent changes of conduction velocity (CV) and conduction block in single A(delta) fibers of primary afferent neurons were characterized in a rat model of neuropathy (NP). Injured dorsal root (DR) fiber in NP rats exhibited profoundly greater decreases of CV following impulse activity than did DR fiber in normal rats. Activity-dependent conduction block was absent up to 100 Hz of activity rate in DR fiber of NP rats, but was present above 25 Hz in normal rats. Profiles of activity dependence in sciatic fibers were similar in both NP and normal rats. These results suggest that nerve injury may alter activity-dependent hypoexcitability of A(delta) DR fibers. Furthermore, this excitability change may be responsible for the elevated pain perception in neuropathy. PMID- 9351644 TI - Relationship between change of movement direction and activity of hippocampal place cells. AB - Hippocampal neurons which increase their activity when the rat passes through a certain position in the environment are called place cells. This paper describes the relationship between their temporal firing patterns and the behavior of the rat with the aim of characterizing the information they encode. The place field was identified in a circular open field during locomotion rewarded by intracranial self-stimulation of lateral hypothalamus. Detailed analysis of temporal firing patterns shows a correlation between changes in the orientation of movement trajectory and occurrence of the last firing of the burst trains emitted during passage of the animal through the place field. This suggests that some hippocampal neurons may encode the spatial information about the orientation changes in the place field. PMID- 9351645 TI - Block of LTP in rat hippocampus in vivo by beta-amyloid precursor protein fragments. AB - The effects of beta-amyloid precursor protein (beta-APP) fragments on plasticity of glutamtatergic synaptic transmission were examined in the hippocampus of urethane anaesthetized rats. I.c.v. injection of beta-amyloid (A beta) 1-40 and 1 42 and the C-terminal fragment CT105 greatly shortened the duration of high frequency stimulation-induced long-term potentiation (LTP) of field excitatory postsynaptic potentials in the CA1 area. Whereas in vehicle injected animals LTP was stable over a 5 h recording period, doses of these peptides (A beta 1-40, 0.4 and 3.5 nmol; A beta1-42, 0.01 nmol; CT105, 0.05 nmol) which did not affect baseline synaptic transmission abolished LTP within 3-5 h. The reduced duration of this form of synaptic plasticity may contribute to the cognitive deficits in Alzheimer's disease. PMID- 9351646 TI - Modulation of rat brain cannabinoid receptors after chronic morphine treatment. AB - Intraperitoneal injection of delta9-THC (7.5 mg/kg) in rats made tolerant to morphine by s.c. implantation of morphine pellets had a much greater analgesic effect than in placebo pellet plus delta9-THC treatment. To investigate whether this was due to some change in cannabinoid receptor levels and/or expression induced by chronic morphine, we designed this autoradiographic binding study coupled with in situ hybridization on sagittal sections of the treated rat brains. Binding showed a significant increase in CB1 receptor density (15%) specifically in the caudate-putamen, in parallel with a significant enhancement of CB1 mRNA in the same area (20%). We suggest that morphine chronic treatment leads to a functional modulation between the opioid and cannabinoid systems at least for analgesia in a specific area, in this case the striatum. PMID- 9351647 TI - Insulin-like growth factor 1 induces climbing fibre re-innervation of the rat cerebellum. AB - The effect of insulin-like growth factor 1 (IGF-1) on neonatal plasticity was studied using the rat olivocerebellar projection as a model. Unilateral removal of climbing fibres in the rat before postnatal day 7 induces re-innervation of the deafferented hemi-cerebellum, which does not occur after postnatal day 10. Rats aged 11 or 12 days underwent climbing fibre transection followed by IGF-1 injection into the denervated cerebellar cortex 24 h later. The exogenous IGF-1 induced climbing fibre re-innervation of the denervated hemicerebellum in a pattern similar to that seen in the immature rat. Thus IGF-1 can extend the window of neonatal plasticity of the brain and therefore may be of potential therapeutic use post-trauma. PMID- 9351648 TI - An CSF anamalous molecular form of acetylcholinesterase in demented and non demented subjects. AB - Analysis of 139 CSF samples from living subjects, using iso-electric focusing in polyacrylamide gels, demonstrated an anomalous molecular form of acetylcholinesterase (AChE). This form was present in 84 of 87 patients with a clinical diagnosis of Alzheimer's disease (AD), 28 of whom have died and in whom histopathological confirmation of AD was obtained. The abnormal AChE form was also present in 22 of 23 patients with clinical dementia not regarded as AD type. In the six patients who died this abnormal AChE form was found in three cases of multi-infarct dementia, one with cerebral glioma with dementia and one with clinical dementia, but no pathology was found based on the Khachaturian criteria for AD. One patient with normal pressure hydrocephalus was negative when tested for the abnormal AChE form. This evidence indicates that the anomalous molecular form of AChE may not be specific for AD, and may possibly be a common indicator for organic dementia. The discovery of this form in 27 of 29 age-matched non demented controls may indicate that the anomalous molecular form of AChE may not only exist in patients with clinically detectable dementia, but is probably present for a period before the onset of dementia. Recognizing and understanding the existence of pre-clinical dementia would be beneficial in designing a strategy for both the prevention and the treatment of dementia. PMID- 9351649 TI - Effects of involuntary auditory attention on visual task performance and brain activity. AB - Involuntary attention to auditory stimulus changes during a visual discrimination task was studied with event-related potentials (ERPs) recorded from the human scalp. A repetitive standard tone or an infrequent, slightly higher deviant tone preceded each visual target stimulus. Deviant tones elicited the mismatch negativity and P3a ERP components and caused increases in reaction time and error rate in the visual task indicating involuntary attention to an auditory stimulus change. These effects were observed even when the tones occurred simultaneously with a visual warning stimulus introduced to keep attention focused on the visual task. In the latter condition, involuntary switching of attention away from the visual task also attenuated the N1 ERP component to visual target stimuli preceded by the deviant tone. PMID- 9351650 TI - Human short latency cortical responses to somatosensory stimulation. A high resolution EEG study. AB - Human short-latency cortical responses to median nerve stimulation were investigated with a new high resolution electroencephalography technology that markedly enhanced spatial details of somatosensory-evoked potentials (SEPs). Maximum amplitude potentials were estimated over contralateral and/or frontal mesial scalp regions about 20, 22, 24, 26, 30, 32 and 45 ms following the stimulation. Frontal-lateral P20-N24-N30-P45 and parietal-lateral N20-P24-P30-N45 showed dipolar patterns, whereas frontal-mesial N24-N30-P45 and central-lateral P22-N26-N32-P45 presented no clearcut dipole counterpart. Plausibly, the spatially enhanced frontal-parietal SEP components were generated (tangential dipoles) within the lateral central sulcus cortex, and anticipated the central lateral and frontal-mesial components generated (radial dipoles) from the crown of the pre- and/or post-central gyri and the supplementary motor area, respectively. PMID- 9351651 TI - Depletion of striatal dopamine transporter does not affect psychostimulant induced locomotor activity. AB - The effect of neurotoxin-induced depletion of striatal dopamine transporter (DAT) binding sites on animals' responses to psychostimulants was investigated. Multiple 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or methamphetamine (METH) injections but not a single METH injection to Swiss Webster mice resulted in > 60% depletion of striatal DAT. MPTP-induced depletion of DAT did not affect METH- and cocaine-stimulated locomotor activity compared with the response of control mice. Pre-exposure to either the neurotoxic or the single non-neurotoxic dose of METH resulted in a marked locomotor sensitization in response to METH or cocaine challenge injections. The present results indicate that > 60% loss in striatal DAT binding sites has no effect on animals' responses to psychostimulants, and suggest that neural systems other than striatal DAT may contribute to the induction of locomotor sensitization to METH and cocaine. PMID- 9351652 TI - Tau phosphorylation in transgenic mice expressing glycogen synthase kinase-3beta transgenes. AB - In order to investigate the effect on tau of manipulating glycogen synthase kinase (GSK)-3beta activity in the brain, we created transgenic mice harbouring wild-type GSK-3beta genes or a mutant GSK-3beta that is predicted to be more active. Transgene-derived mRNAs were detected in the brains of a number of the transgenic mouse lines and several of these transgenic lines displayed transgenic GSK-3beta activity. Western blot analyses of the two lines with the highest levels of transgenic GSK-3beta activity revealed that the phosphorylation status of tau was elevated at the AT8 epitope. These observations strongly suggest that GSK-3beta is an in vivo tau kinase in the brain. Only low levels of expression of GSK-3beta were obtained and it is possible that high levels of GSK-3beta activity are lethal. PMID- 9351653 TI - Serotonin inhibits calcium-activated K+ current in rat taste receptor cells. AB - Little is definitively known of the identity or actions of neurotransmitters utilized within mammalian taste buds. Serotonin has been immunocytochemically localized to taste cells of several species but its physiological actions are unknown. Using whole-cell patch clamp recordings on dissociated posterior rat taste cells, data are presented to suggest that exogenously applied serotonin inhibits a calcium-activated potassium current by up to 50%. This current, best visualized at depolarized holding potentials, is both apamin- and charybdotoxin sensitive. Approximately 60% of the tested taste cells were serotonin sensitive. This inhibition was mimicked by N-(3-trifluoromethylphenyl)piperazine (TFMPP), a general serotonin receptor agonist, by 8-hydroxy-dipropylaminotetralin (8-OH DPAT), a selective 5-HT1A receptor agonist, but not by phenylbiguanide, a 5-HT3 receptor agonist. These are the first data to establish a physiological effect of serotonin on mammalian taste cells. PMID- 9351654 TI - Cycloheximide phase-shifts, but does not prevent, de novo Krox-24 protein expression. AB - Previous studies show that focal hippocampal injury transiently increases NMDA receptor-dependent expression of inducible transcription factors (ITFs including Krox-24) in rat dentate gyrus neurons. Furthermore, pretreatment with the protein synthesis inhibitor, cycloheximide (CHX), prevents de novo ITF protein expression 1 h post-injury. Here, we further characterize the effects of a single pretreatment dose of CHX on injury-induced expression of Krox-24 and show that CHX pretreatment phase-shifts (delays), but does not prevent, de novo expression of Krox-24 in hippocampal dentate gyrus neurons following injury. This may have implications for studies which use CHX pretreatment to examine the role of gene expression and de novo protein synthesis in long-term memory formation, the stabilization of long-term potentiation, kindling and neuronal injury. PMID- 9351655 TI - Sensitization of postsynaptic dorsal column neuronal responses by colon inflammation. AB - The role of a newly identified component of the postsynaptic dorsal column (PSDC) system in viscerosensory processing has been recently described. The purpose of this study was to examine the effect of colon inflammation on the responses of single PSDC cells, located in the vicinity of the central canal at L6-S1 spinal segments, to graded colorectal distension (CRD) and to cutaneous stimulation. Experiments were conducted on seven male Sprague-Dawley rats anesthetized with pentobarbital. Recordings were made from seven PSDC cells located around the central canal at L6-S1 in response to CRD and cutaneous stimulation before and after colon inflammation. Inflammation of the colon with mustard oil (MO) induced an increase in the background activity of these cells. Colon inflammation also potentiated the responses of the PSDC cells to graded CRD but not to cutaneous stimulation. This is consistent with previously observed effects of colon inflammation on the responses of viscerosensitive cells in the ventral posterolateral (VPL) nucleus of the thalamus and in the nucleus gracilis (NG). These observations support a role of the PSDC system in viscerosensory processing and primary visceral hyperalgesia. PMID- 9351656 TI - Obligatory role of the LIFG in synonym generation: evidence from PET and cortical stimulation. AB - We report results from a patient in whom we obtained converging evidence from positron emission tomography (PET) and intraoperative stimulation mapping to support a one-way dissociation between the functional areas involved in word repetition and synonym generation. Intraoperative stimulation mapping interfered with synonym generation but did not disturb word repetition at the same left inferior frontal site at which a cerebral blood flow (CBF) increase had been observed for a synonym generation task. The results for this single subject suggest that the functional areas involved in different aspects of linguistic processing are dissociable and that specific disruption under conditions of cortical stimulation can be correlated with the brain regions identified via PET as the most active during performance of a specific task. PMID- 9351657 TI - Recognition memory for words and pictures: an event-related potential study. AB - Event-related potentials (ERPs) were recorded during the test phase of recognition memory tests for words and pictures of objects. ERPs elicited by recognized items contained a temporo-parietally distributed positive shift (the parietal old/new effect), which was strongly left lateralized regardless of stimulus type. This finding suggests that the lateral distribution of the parietal old/new effect is unrelated to the lateralization of the memory functions supported by the medial temporal lobe memory system. The ERPs to pictures, but not to words, also demonstrated frontally distributed old/new effects, which shifted over time from a left- to a right-sided maximum. These effects may reflect the richer informational content associated with episodic memory for pictures. PMID- 9351658 TI - Synaptic inputs on rat brainstem motoneurones in organotypic slice culture. AB - To study the formation of target specific afferents on brain stem motoneurones of the rat, we used an organotypic co-culture of embryonic rat (E18) brain stem explants containing the facial or hypoglossal motor nuclei together with a tongue explant. The brain stem explants also contained known dorsal premotor structures such as lateral reticular nuclei and vestibular or spinal trigeminal nuclei. In cultures maintained in vitro for over 3 weeks, silver impregnation studies identified neurones in the dorsal sensory structures with axons arborizing within the motor nucleus. A double fluorescent labelling procedure demonstrated that axons originating from dorsal sensory regions come in close contact with identified motoneurones. Electrical stimulation of neurones in the dorsal regions induced monosynaptic and polysynaptic EPSPs and spikes in identified motoneurones together with muscle contraction. This work demonstrates that premotor structures in slice cultures develop organotypic functional synaptic connections with embryonic brain stem motoneurones. PMID- 9351659 TI - Opposite effects of lanthanum on different types of nicotinic acetylcholine receptors. AB - The effects of lanthanum (La3+) were studied on muscle and neuronal nicotinic acetylcholine receptors (AChRs) expressed in Xenopus oocytes. La3+ exerts a dose dependent positive modulation on alpha1 beta1 gamma8 muscle AChRs, whereas it modulates negatively either alpha2 beta2, alpha2 beta4 or alpha3 beta4 neuronal AChRs. Moreover, La3+ appears to accelerate the desensitization of neuronal receptors. In both muscle and neuronal AChRs, the respective potentiating or inhibiting effects of La3+ on the ACh-currents are voltage-independent, suggesting that La3+ is acting at a site located in the external domain of the receptor. PMID- 9351660 TI - The beta-amyloid epitope masking activity in human brain is identified as albumin. AB - Human brain homogenate proteins were analyzed for binding and processing activity in relation to brain beta-amyloid precursor protein (APP). The homogenate was purified by arginine-Sepharose 4B affinity chromatography, which traps proteins with affinity to certain groups of arginine residue, such as serine proteases and zymogens. A 69 kDa protein that masks epitope(s) of brain APP was found in a weakly bound fraction. The nature of the 69 kDa brain protein was identified as albumin by N-terminal amino acid sequencing and Western blot analysis using anti human albumin antibody. Western blot analysis with domain-specific anti-APP antibodies revealed that the masking activity is complete for beta-amyloid epitope(s), but incomplete for cytoplasmic and extracellular domain epitopes, suggesting that the interaction site of the albumin is beta-amyloid itself. Therefore, it seems that brain albumin is not merely a carrier protein for beta amyloid in cerebrospinal fluid, but also a modulator which interferes with processing of beta-amyloid precursor protein and its peptides. PMID- 9351661 TI - Marked suppression of cortical auditory evoked response shortly before the onset of REM sleep. AB - In 10 of 12 subjects examined, the amplitude of N300, a component of the cortical auditory evoked potential, was evidently smaller in rapid eye movement (REM) sleep than in non-REM sleep. The start of the reduction associated with the onset of the first episode of REM sleep was examined in these 10 subjects. In five of these, a marked reduction of N300 amplitude occurred 0.5-2.5 min before the appearance of muscle atonia of REM sleep. In two subjects, a similarly marked reduction of the N300 amplitude occurred 0.5-1.0 min before the disappearance of sleep spindles or K-complexes. This suggests that a suppression of the synchronizing mechanism in the cerebrum sometimes occurs briefly prior to the occurrence of other physiological phenomena associated with REM sleep. PMID- 9351662 TI - Regulation of GDNF expression in cultured astrocytes by inflammatory stimuli. AB - Astrocytes express increased levels of neurotrophic factors in response to pathological conditions in the CNS such as injury and inflammation. We have examined the effects of lipopolysaccharide (LPS) and inflammatory cytokines on the expression of GDNF by mouse astrocytes and by C6 glial cells. LPS and tumor necrosis factor-alpha (TNF-alpha) induced an increase in level of glial-derived neurotrophic factor (GDNF) mRNA in both cell types. Similarly, the synthesis of GDNF protein was increased by both treatments. Interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma) induced similar effects on GDNF production, whereas IL-2 and IL-6 had no significant effects. These results indicate that the expression of GDNF in astrocytes is regulated by inflammatory stimuli and therefore may provide neurotrophic support to injured neurons in inflammatory conditions in the CNS. PMID- 9351664 TI - Multiple transcripts encode the 5-HT1F receptor in rodent brain. AB - The mouse serotonin 1F (5-HT1F) receptor is encoded by at least three transcripts in mouse brain. These transcripts are expressed predominantly in cortex and hippocampus. Similar transcripts are seen in Northern analysis of rat brain mRNA. 5' RACE showed a predominant transcription start site around 350 bp upstream of the translational start present in mouse cDNA. Our results suggest that the heterogeneity seen in transcript size is due to differences in the 3' untranslated region, which could play a critical role in mRNA targeting and localization. The mouse 5-HT1F genomic clone shows the coding region to be intronless and an intron splice junction is seen in the 5' untranslated region which is conserved in both rat and mouse. PMID- 9351663 TI - Expression of peripherin, NADPH-diaphorase and NOS in the adult rat neocortex. AB - Peripherin is mainly expressed in peripheral neurones and in CNS neurones which extend axons into peripheral nerves. However, this intermediate filament protein has also been detected in a few other neurones entirely located within the CNS. The present study focuses on the adult rat neocortex. Peripherin immunoreactive (P+) neuronal somata and their neuritic extensions were observed in cortical layers II, III, V and VI, while a few P+ nerve fibres could be seen in layer I. All the P+ neurones could be selectively stained using reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry, a typical feature of aspiny neurones. Some of the P+ neurones could also be immunostained with an antibody raised against nitric oxide synthase (NOS). These results provide evidence that peripherin is present in a discrete population of aspiny interneurones of the adult rat neocortex. The functional significance of the co expression of peripherin and NOS needs further investigation. PMID- 9351665 TI - Comparison of the neuroprotective effects of APV and bcl-2 in glutamate-induced cell death. AB - The neuroprotective effects of the NMDA receptor blocker APV were compared with those of bcl-2 in a glutamate-induced excitotoxic cell death model in cultured rat cortical neurons. Exposure to 100 microM glutamate for 5 h caused approximately 95% of the cultured neurons to die in 24 h. The NMDA-selective antagonist D-(-)-2-amino-5-phosphonopentanoate (APV) protected the neurons effectively when applied prior to or soon after glutamate treatment. However, infection with a viral vector expressing the proto-oncogene bcl-2 strongly protected neurons even if applied as late as 8 h following the glutamate insult. These data provides evidence that APV blocks an early stage of the death cascade in response to elevations of glutamate. By contrast bcl-2 appears to act at a fairly late stage in the cell death process and these results suggest a possible clinical role in treatment of ischemic brain disorders. PMID- 9351667 TI - Okadaic acid modulates the cytoskeleton changes induced by amyloid peptide (25 35) in cultured astrocytes. AB - Amyloid beta-protein (25-35) (betaA) induced a marked morphological change in astrocytes, changing their flat polygonal shape into a stellate process-bearing morphology. The changes induced by betaA were concentration and time-dependent, whereas the addition of a scrambled peptide did not alter astrocyte morphology. We discard the possibility of betaA-astrocytes being type II-like astrocytes. We also analysed the influence of the presence of kinase and phosphate inhibitors on this morphological change. Our data indicate that the betaA-induced phenotype was not affected by the inhibition of protein tyrosine kinase or tyrosine phosphatases. Only the addition of okadaic acid to astrocytes prevented the morphological transformation from flat to stellate shape, induced by betaA (25 35). Inhibition of the stellate phenotype by okadaic acid was initiated at a concentration of 10 nM which suggested that either phosphatase 2A or 1 plays an important role in the betaA astrocytic transformation. PMID- 9351666 TI - Dopamine transporter mRNA levels are high in midbrain neurons vulnerable to MPTP. AB - The neurotoxin MPTP kills only certain midbrain dopaminergic (DA) neurons to produce a model of Parkinson's disease. The dopamine transporter (DAT) is important to MPTP toxicity because to be neurotoxic, an MPTP metabolite must first gain access to the DA neuron via the DAT. Also, MPTP is less toxic to DA neurons that contain the putative neuroprotective calcium-binding protein calbindin-D28k (CB). The present study examined the relative importance of DAT activity and CB for cellular vulnerability to MPTP-induced degeneration in the C57BL/6 mouse. Cells that were vulnerable to MPTP were found to contain high levels of DAT mRNA, whereas cells that were not vulnerable contained low levels. Also, the few substantia nigra cells remaining after a toxic dose of MPTP contained only low levels of DAT mRNA. However, there was not a strong relationship between cellular resistance to MPTP toxicity and cells containing CB. These data provide in vivo evidence for a direct correlation between midbrain cellular vulnerability to MPTP toxicity and the activity of the DAT. PMID- 9351668 TI - Computer model of clonazepam's effect in thalamic slice. AB - In the thalamus, paradoxical changes in response to augmentation of inhibition can occur as a result of either cellular or network effects. Clonazepam, a GABA(A) agonist, produces a paradoxical reduction in evoked thalamocortical neuron inhibitory postsynaptic potential (IPSP) in thalamic slice. This has been hypothesized to be a result of augmentation in inhibitory to inhibitory connections. In a computer model, orthodromic simulation produced an increase in initial IPSP, a result contrary to that found experimentally. This failure was traced to the inability of orthodromic activation to produce fast enough recurrent inhibition to alter initial reticularis neuron firing. Simulated antidromic stimulation was able to reduce this initial spike train and reproduced the experimental finding. PMID- 9351669 TI - Cellular mechanism for the temperature sensitive spatial orientation in Clione. AB - The swimming mollusk Clione is normally oriented vertically. As water is warmed, this orientation is lost or reversed. CPB3 interneurons, which transmit signals from the statocyst receptors (SRCs) to the tail motoneurons and play a key role in space orientation, were strongly depolarized upon warming. Normally, intracellular stimulation of the rostro-dorsal SRC (DSRC) excited CPB3b. Upon warming the excitation gradually decreased and in some cases was even replaced by inhibition. The reversal potential for the synaptic potentials (PSP) produced in CPB3b by DSRC stimulation is depolarized relative to the normal membrane potential at lower temperature. Warming causes depolarization of the membrane potential such that the PSP reversal potential is approached and even passed, with attenuant effects on PSP amplitude and polarity. This effect provides a mechanism for the temperature sensitive changes in the orientation of Clione. PMID- 9351670 TI - Localization of mGluR4 protein in the rat cerebral cortex and hippocampus. AB - The cellular distribution of the rat metabotropic glutamate receptor type 4 (mGluR4) was examined in the adult rat cerebral cortex and hippocampus. Antibodies were raised against amino acid residues located in the extracellular amino terminal domain that is common to both the mGluR4a and mGluR4b splice variants, and used for an immunohistochemical investigation. The affinity purified antibodies on immunoblot analysis specifically detected mGluR4 protein in transfected mammalian cells, showing no cross-reactivity with other members of the mGluR family. At the light microscope level intense mGluR4-like immunoreactivity was detected in the CA2 region of Ammon's horn in the hippocampus. In the cerebral cortex numerous non-pyramidal cells were strongly immunolabelled. PMID- 9351671 TI - Blockade of nitric oxide-evoked smooth muscle contractions by an inhibitor of guanylyl cyclase. AB - Nitric oxide-induced contractile responses of smooth muscle were studied in vitro in guinea-pig small intestine. Application of nitric oxide (NO; 0.3-30 microM) evoked a small initial relaxation followed by a marked contractile response in plexus-containing longitudinal smooth muscle preparations from small intestine. The extent of the NO-evoked contractile response was dose-dependent and the response was blocked by tetrodotoxin. Atropine significantly reduced the NO evoked contraction and the remaining part was abolished by the NK1-receptor antagonist CP 96,345. An inhibitor of soluble guanylyl cyclase, ODQ (1H [1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one), abolished the NO-evoked contractile response. The results suggest that NO, in addition to the classical direct smooth muscle relaxing effect, causes activation of excitatory neurones, via a pathway utilizing soluble guanylyl cyclase, which leads to a smooth muscle contraction. PMID- 9351672 TI - Post-hypoxic hypothermia reduces cerebrocortical release of NO and excitotoxins. AB - Hypothermia applied after hypoxia offers neuroprotection in neonatal animals, but the mechanisms involved remain unknown. Hypoxia was induced in newborn piglets and changes in excitatory amino acids (EAAs) and the citrulline:arginine ratio (CAR) were followed by microdialysis for 5 h. After the 45 min hypoxic insult, the animals were randomized to receive normothermia (39 degrees C; n=7) or hypothermia (35 degrees C; n = 7). After reoxygenation, extracellular glutamate, aspartate and the excitotoxic index were significantly lower in the cerebral cortex of hypothermic animals than in normothermic animals. A progressive rise of the CAR occurred during reoxygenation in the normothermic group whereas the ratio tended to decrease in the hypothermic group. In conclusion, post-hypoxic hypothermia attenuated NO production and overflow of EAAs. PMID- 9351673 TI - Auditory evoked magnetic fields in patients with right hemisphere language dominance. AB - Auditory evoked magnetic fields for pure-tone stimuli were measured in seven subjects with right hemisphere dominance for language using a whole-head magnetoencephalography system linked to magnetic resonance imaging. N100m responses were observed in both hemispheres in five subjects. The N100m response latency to contralateral stimulation was significantly shorter on the right than on the left in all cases. Normal right-handed subjects with left hemisphere dominance showed exactly the same responses. Therefore, the shorter N100m latency in the right hemisphere is independent of the dominant hemisphere for language. PMID- 9351674 TI - CNS GABA neurons express the mu-opioid receptor: immunocytochemical studies. AB - It has been proposed that mu-opioid receptors excite neurons in hippocampus and nucleus raphe dorsalis (NRD) by decreasing GABAergic tone. In the present study, we examined whether immunocytochemical evidence of interaction between GABAergic neurons and the mu-opioid receptor could be found in the CNS. Portions of rat brain were sectioned and stained for GABA and for the cloned mu-opioid receptor (MOR1) using two-color immunofluorescence. Neurons double-labeled for GABA and MOR1 were present in hippocampus and NRD, as well as in olfactory bulb, dorsal lateral periaqueductal gray matter, nucleus raphe medianis, nucleus raphe obscurus, and the spinal trigeminal nucleus and tract. We conclude that expression of the mu-opioid receptor by GABAergic neurons is common in the rat CNS. PMID- 9351675 TI - Item and source memory: differential age effects revealed by event-related potentials. AB - The neural substrates of age-related memory differences were evaluated by recording event-related potentials (ERPs) from young and older adults during a recognition memory paradigm. Subjects studied two temporally distinct lists of sentences (each with two nouns) and were tested for their memory of the nouns and of the list (i.e. temporal source) in which they had occurred. Compared with the young, the old showed a greater source than item memory performance decrement. Both age groups showed equivalent posterior-maximal old/new ERP effects. However, only the young produced a frontal-maximal, late onset old/new effect that differed as a function of subsequent source attribution. Age-related explicit memory differences may be due to a deficit in a prefrontal cortical system that underlies source memory. PMID- 9351677 TI - Attenuated glutamate release during ischemia in ethanol-administered gerbils. AB - Previous studies have found an association between prior ethanol consumption and aggravated stroke outcome. Gerbils were intermittently given ethanol injections (s.c.) for 21 days at doses of 1 and 4 g/kg. After cessation of injections and appropriate weight gain, subjects underwent bilateral carotid occlusion while amino acid neurotransmitter levels in the hippocampus were monitored. Both the low and high dose ethanol groups demonstrated significantly decreased glutamate release compared with saline-treated controls during ischemia (p < 0.05). These results are consistent with a long-lasting ethanol-induced decrease in synaptic density in the hippocampus. That no intergroup differences on histological or neurobehavioral measures was found may suggest a functional dissociation of glutaminergic involvement in the pathogenesis of aggravated stroke outcome with alcoholism. PMID- 9351676 TI - Alterations of GABA(A)beta2/3 immunoreactivity in the dentate gyrus after perforant pathway lesion. AB - Immunocytochemical techniques were employed to examine the changes in the GABA receptor subunits beta2/3 within the dentate gyrus of the rat brain 1, 3, 7, 14, 30 and 90 days after a unilateral perforant pathway lesion. Three days post lesion we observed a decrease in beta2/3 immunolabeling in the inner molecular layer of the dentate gyrus followed by a comparable decrease in the outer molecular layer 7 days post-lesion. These decreases were transient; 30 and 90 days post-lesion, beta2/3 immunolabeling appeared similar to controls in the inner portion of the molecular layer, while in the outer region the labeling was increased. In this latter region we also observed a dense band of AChE fibers. Following survival times of 3 days we observed a diffuse staining of the neuropil in the hilar region, and a dense amorphous accumulation of peroxidase reaction product in the polymorphic region. These responses were transient and by 14 days the hilar/polymorphic region appeared indistinguishable from controls. These data suggest a unique pattern of immunoabeling in the molecular and polymorphic region in response to perforant pathway lesion. A putative explanation for this response is discussed. PMID- 9351678 TI - NOS type-1 mRNA expression and protein localization in spinal autonomic neurons. AB - Autonomic neurons of the rat spinal cord show strong NADPH diaphorase activity and immunoreactivity for nitric oxide synthase (NOS). Here we show mRNA expression of NOS type-1 (neuronal or brain NOS) transcripts in cell bodies of sympathetic preganglionic neurons (SPNs) of the intermediolateral (IML) cell column by non-radioactive in situ hybridization using NOS-I riboprobes. Hybridization signals occurred only in neuronal cell bodies and not outside, in what appeared to be fibers and/or terminals. In preganglionic fibers of SPNs, however, dense axoplasmic immunogold labeling was detected with a monoclonal anti NOS-I antibody. Expression of NOS-I mRNA in SPN cell bodies and axoplasmic immunolocalization of NOS-I protein suggest that protein translocation is involved in NO-mediated preganglionic control of peripheral targets. PMID- 9351679 TI - Vestibular inputs to bulbar respiratory interneurons in the cat. AB - Vestibular inputs to medullary respiratory interneurons were studied in decerebrated and artificially ventilated cats. Extracellular recordings were made from 40 neurons located in the area of pre-Botzinger complex and activated antidromically from the contralateral ventral respiratory group. Neuronal populations analyzed included inspiratory and expiratory neurons with augmenting, constant and decrementing firing patterns, and a late inspiratory neuron. Seventeen neurons responded to ipsilateral and/or contralateral vestibular nerve electrical stimulation. These responses were observed in all seven cell types. Most neuronal reflex responses consisted of inhibition, while a few consisted of either excitation or a combination of both inhibition and excitation. These results indicate that pre-Botzinger respiratory interneurons, which may be involved in respiratory rhythmogenesis, also participate in vestibulorespiratory responses. PMID- 9351680 TI - Subarachnoid hemorrhage induces c-fos, c-jun and hsp70 mRNA expression in rat brain. AB - To detect stress responses of the brain to subarachnoid hemorrhage (SAH), we investigated the expression of immediate early genes (IEGs) and hsp70 mRNA by in situ hybridization. Experimental SAH was produced in 49 rats by endovascular penetration. We also monitored the intracranial pressure (ICP) changes. The genes c-fos and c-jun were induced in the cerebral cortex, hippocampus and dentate gyrus in the penetrated side. mRNA coding for hsp70 was induced in the cerebral cortex, hippocampus, thalamus, hypothalamus and caudoputamen in the penetrated side and extended to the contralateral hemisphere. IEGs in the cerebral cortex were completely blocked by MK-801 pretreatment, but hsp70 mRNA was not. This suggests that the expression of IEGs correlates with spreading depression. The IEGs and hsp70 expression may reflect the severity of SAH impact and relate to the mechanisms of symptomatic vasospasm. PMID- 9351682 TI - ATP-promoted amyloidosis of an amyloid beta peptide. AB - Amyloidosis is implicated in the aetiology of a number of disorders of human health. The factors that influence its instigation and subsequent rate of progress are the subject of a considerable research effort. The peptide fragment A beta(25-35) is amyloidogenic and has proven to be a useful model of the processes involved in amyloidosis. It is demonstrated herein that the assembly of A beta(25-35) into thioflavin T-reactive fibrils and their subsequent rearrangement into advanced glycation endproducts is accelerated by ATP. Aluminium potentiated these effects of ATP, suggesting a possible link with the aetiology of amyloidoses in vivo. PMID- 9351681 TI - Auditory projections from the IC to the SCN by way of the LG in the mole, Mogera. AB - To study morphological substrates for sensory specialization in subterranean mammals, we investigated both auditory and visual pathways in the mole. The inferior colliculus (IC), an auditory relay, projects not only to the medial geniculate, the major gateway to the auditory cortex, but also to the lateral geniculate (LG), the major gateway to visual cortex. Further evidence is that the LG does not send many fibers to the cortex in the mole. Instead, the auditory inputs to the LG are likely to be conveyed to the suprachiasmatic hypothalamic nucleus (SCN), which plays a role in photoperiodic functions in common mammals. Auditory inputs to the SCN may subserve periodic reproductive behaviors in the exclusively separated territorial domains of subterranean mammals. PMID- 9351683 TI - Intracellular responses of the rat cochlear nucleus to sound and its role in temporal coding. AB - The anteroventral cochlear nucleus (AVCN), the first centre of the central auditory pathway, contains globular bushy cells, which are unique in their ability to produce fast excitatory post-synaptic potentials (EPSPs). Using in vivo intracellular recordings in the rat AVCN we examined these fast EPSPs in relation to temporal coding. At frequencies up to 2.5 kHz, EPSPs were evoked on successive sine waves of the stimulus with EPSP summation limited. This one-to one relationship between the EPSPs and the sound wave period was present at higher frequencies and over a greater intensity range than for action potentials. These results suggest that temporal coding is possible in globular bushy neurones by their ability to extract temporal information through fast processing of convergent presynaptic input. PMID- 9351684 TI - Effect of step duration during incremental exercise on breathing pattern and mouth occlusion pressure. AB - We compared the effects of two step durations on breathing pattern, mouth occlusion pressure and "effective" impedance of the respiratory system during incremental exercise. Nine normal subjects (mean age: 27.8+/-1.21 years) performed two incremental exercise tests in randomized order: one test with step increments every 1 min 30s and the other, every 4 min. After a warm-up at 25 W for the 1 min 30 s test, the power was increased by 50 W from 50 W to exhaustion. During the last minute at each power, we measured ventilation (VE), tidal volume (VT), breathing frequency (fR), inspiratory and expiratory time (TI and TE), total time of the respiratory cycle (TTOT), TI/TTOT, mean inspiratory flow (VT/TI), mouth occlusion pressure (P0.1), "effective" impedance of the respiratory system (P0.1/(VT/ TI)) and venous blood lactate concentration ([La]). Our result showed that at maximal exercise the power was significantly higher (p < 0.01) and [La] lower (p < 0.01) in the 1 min 30 s test. At 100, 150 and 200 W, the 4 min test showed significantly higher oxygen uptake (VO2), carbon dioxide output (VCO2), VE, P0.1, fR, VT/TI and HR (p <0.001) and significantly lower TI, TE and TTOT (p<0.01). [La] was significantly higher at 150 W (p<0.05) and 200 W (p<0.001). At the same VCO2, P0.1 was not significantly different between the two tests, whereas VE showed a tendency to be higher (p = 0.08) and P0.1/(VT/TI) was significantly lower during the 4 min test. In conclusion, this study allowed us to quantify the difference in inspiratory neuromuscular output and ventilatory response between 1 min 30s and 4 min tests and showed that different step durations alter the relationship between inspiratory neuromuscular output and mean inspiratory flow. PMID- 9351685 TI - Bone density and bone metabolic markers in active collegiate athletes: findings in long-distance runners, judoists, and swimmers. AB - We investigated the bone metabolic system status of 103 male and female volunteer collegiate athletes, who were actively pursuing one of three different sports: Long-distance running (LR); judo (JU); and swimming (SW). The following parameters were evaluated: total body bone mineral density (TMBD); bone-forming metabolic markers; serum procollagen type I C-peptide (PICP) levels; bone alkaline phosphatase (B-ALP) content; bone resorption markers, urinary pyridinoline (Pyd) and deoxypyridinoline (Dpd) levels. We found that the TBMD and urinary Dpd values in JU athletes were significantly higher (p < 0.001) than in athletes of the same sex in the other two groups. The urinary Pyd level in male JU athletes was also higher (p < 0.001) than that in the other two groups, but that in females JU athletes was only higher (p < 0.01) than that in female LR athletes. The PICP levels were similar to the TBMD values in all groups. No differences in bone density or in bone metabolic markers were seen in LR and SW athletes of the same sex. We thus conclude that differences in bone mineral density are in part due to the demands of the specific sport, and that they are reflected in bone metabolic markers. In addition, the status of bone metabolic turnover in male JU athletes in training may be hypermetabolic and as well as that of female JU athletes with regular menses cycles. PMID- 9351686 TI - Skeletal maturation, somatic growth and physical fitness in girls 6-16 years of age. AB - The importance of chronological age (CA) and skeletal age (SA) in explaining variation in somatic dimensions, and the independent contributions of CA, SA, stature (ST) and weight (WT) to variability in physical fitness were investigated in a sample of 6593 girls 6-16 years of age. Body dimensions included lengths, breadths, circumferences, skinfolds, and Heath-Carter somatotype, while fitness tests included measures of health- and performance-related fitness, and cardiovascular and lung functions. Age-specific correlations were calculated between SA and anthropometric dimensions, fitness tests and cardiovascular and lung functions, while age-specific stepwise multiple regressions were used to investigate the relative importance of SA, CA, ST and WT in explaining fitness and cardiovascular and lung functions. SA is most highly correlated with lengths and then with breadths, circumferences and skinfolds in this order. SA per se or in interaction with CA is the only significant predictor of somatic characteristics. Among fitness items, physical working capacity and static strength correlate highest with SA. Bent arm hang, leg lifts and sit-ups correlate negatively with SA but values are low, while all other components correlate at non-significant or low levels. Results of the multiple regression analysis indicate that, with few exceptions, CA, SA, ST and WT and their interactions explain less than 10% of the variance in most physical fitness items. However, for PWC, arm pull strength, and bent arm hang, the interaction terms explain between 12% and 67% of the variance. PMID- 9351687 TI - Gender effects on submaximal energy expenditure in children. AB - Previous studies have suggested that submaximal energy expenditure relative to body mass during weight-bearing exercise may be greater in boys compared to girls. This two-part study examined a) gender-related five-year longitudinal changes in submaximal walking economy and b) gross, net, and delta muscle work efficiency during submaximal cycle exercise in a cross-sectional analysis of boys and girls. In the longitudinal study, the influence of pre-exercise metabolic expenditure, stride frequency, and substrate utilization (by RER) on changes in economy were examined. During the five years, mean VO2 per kg during submaximal treadmill walking (measured at 8% slope, 3.25 mph) decreased 16% in girls and 13% in boys (p > 0.05 for gender). Likewise, no significant gender differences were observed in decline of stride frequency over time. RER values were similar between sexes except in the final two years when girls had significantly greater values than the boys. No gender-related differences were observed in any measure of muscle work efficiency. This study failed to reveal significant gender differences in utilization of energy during submaximal exercise in children. PMID- 9351688 TI - Exercise training of moderate intensity does not abate the effects of denervation on muscle morphology. AB - Denervation elicits profound alterations in the morphometry of skeletal muscle. These alterations include changes in fiber type composition as well as reductions in fiber size. There is evidence that the increased mechanical load placed upon muscle via rhythmic stretching attenuates denervation induced alterations in muscle morphology. The purpose of the present study was to determine whether the mechanical stimuli associated with exercise training, i.e. rhythmic stretching and mechanical loading, would effectively moderate the changes in muscle morphometry observed following denervation. Unilateral denervation of the soleus muscle of eight male Sprague-Dawley rats was performed under aseptic conditions. The animals were then randomly assigned to two groups: sedentary controls and exercise trained. The exercise training protocol featured treadmill running five days per week for six weeks. At the conclusion of the experimental period, animals from both groups were sacrificed and soleus muscles were histochemically analyzed for muscle morphometry. Results demonstrated that denervation caused marked alterations in fiber type profile and in fiber cross-sectional areas. Interestingly, the degree of denervation induced atrophy appeared to be fiber type specific. However, the data presented here indicate that in denervated soleus muscles there were no significant differences in fiber type composition or fiber size between the sedentary and exercise trained groups. Hence, it appears that the mechanical stimuli provided by treadmill running of moderate intensity and duration are not sufficiently potent to ameliorate muscle morphometric responses to denervation. PMID- 9351689 TI - The effect of training status on the serum creatine kinase response, soreness and muscle function following resistance exercise. AB - Untrained individuals develop muscle soreness and increased serum creatine kinase (CK) activity in the blood after strenuous, unaccustomed exercise. An unpublished observation in our laboratory revealed that trained weightlifters also experience considerable soreness after unaccustomed exercise, but may not show a dramatic CK response. This study examined the CK and soreness responses to strenuous exercise in weightlifters (TR, n = 10) and untrained subjects (UTR, n = 10). Trained subjects had a minimum of three years weightlifting experience, and regularly performed squats and leg presses. Untrained subjects had not participated in any regular resistance exercise for the past three years. Following two acclimation sessions, subjects reported to the lab on seven consecutive days and on the tenth day after knee extensor exercise. Weight training sessions occurred on day 1 for the knee extensors (KE) and day 2 for the knee flexors (KF). The weight training consisted of these exercises (sets): squat (5), leg press (3), leg extension and lunge (3) for the KE, double leg curls (6), single leg curls (3), stiff-legged deadlifts (4, TR group only) for the KF at 12 RM for all exercises. To document the stress due to exercise, the loss in strength (isometric peak torque, IPT) was assessed on a Biodex isokinetic dynamometer. Maximal voluntary IPT of the KE at 90 degrees and the KF at 80 degrees decreased 17-30% with no significant differences between groups. Muscle soreness during simulated squat leg curl movement was assessed by a 100 mm visual analog scale (VAS). Average peak KE soreness was 76 mm for TR and 58 mm for UTR, KF soreness was 60 mm for TR and 47 mm for UTR post-exercise. Serum CK levels were significantly different between groups with a peak of 1349 IU for TR and 3272 IU for the UTR (p < 0.01). Although the TR group experienced greater soreness than the UTR, peak serum CK activity was significantly lower, suggesting that trained individuals can develop severe soreness without the same degree of increase in serum CK activity observed in untrained individuals. PMID- 9351690 TI - In vitro model of characterizing the effects of compressive loading on proteoglycans in anatomically intact articular cartilage. AB - An in vitro method has been developed of cultivating anatomically intact articular cartilage (humeral head of dog) while excluding both bone tissue and other connective tissues with a condom, which controls changes in hydration and minimizes loss of proteoglycans. Compressive loading experiments were carried out with the condom-covered humeral head, to which contact stresses of 2.1 MPa, 3.3 MPa and 6.4 MPa, respectively, were applied with a rubber disc of a simple loading apparatus. The model makes it possible, to compare experimentally loaded regions with experimentally unloaded regions in both the same joint area and the contralateral control joint. Intermittent pressure loading (4-s-on/16-s-off cycle) during 2 hours of pulse-experiment and 18 hours of chase-experiment resulted in a 40% increase in proteoglycan synthesis rate at 2.1 MPa, an unaltered synthesis rate at 3.3 MPa, and a 45% decrease in proteoglycan synthesis rate at 6.4 MPa, as measured by 35S-sulfate incorporation. No significant increase in degradation rate of proteoglycans was noted during the various loading experiments. PMID- 9351691 TI - The Cosmed K4 telemetry system as an accurate device for oxygen uptake measurements during exercise. AB - The purpose of this study was to test the accuracy of oxygen uptake (VO2) measurements using the Cosmed K4 portable telemetry system. This system of higher technology than the original Cosmed K2 device, contains a CO2 electrode allowing measurements alternatively by either the Cosmed K4 system (K4) or the CPX Medical Graphics (CPX) during a maximum oxygen uptake (VO2max) ergocycle test, at rest and during several submaximal exercises (25, 50 and 75% of maximal work rate) in seven subjects. Heart rate values were comparable for exercise at the same work stage during gas collection using the two systems, indicating that the physiological stresses were similar. The VO2 values did not significantly differ at rest (4.40+/-0.83 vs 4.16+/-0.58ml x min(-1) x kg[-1]), at 25% Wmax (20.97+/ 1.31 vs 21.32+/-2.54 ml x min(-1) x kg[-1]), at 50% Wmax (33.32+/-3.92 vs 33.50+/ 3.51 ml x min(-1) x kg[-1]), at 75% Wmax (47.01+/-7.51 vs 47.49+/-7.11 ml x min( 1) kg[-1]) and at maximal intensities (62.07+/-8.48 vs 62.84+/-11.31 ml x min(-1) kg[-1]) using K4 and CPX devices, respectively. The results of this study indicated that the K4 system was accurate for all oxygen uptake measurements from rest to maximum exercise levels. PMID- 9351692 TI - The validity of the telemetric system CORTEX X1 in the ventilatory and gas exchange measurement during exercise. AB - With the portable spirograph CORTEX X1 both the oxygen consumption and the carbon dioxide output can be determined. Therefore, the aim of the present study was to determine the accuracy of the CORTEX X1 in measuring F(E)O2, F(E)CO2 and V(E) when attached to a motor-driven mechanical syringe and to validate the CORTEX X1 against a standardized breath-by-breath system during a graded bicycle ergometry. Fifteen subjects (8 male, 7 female; 26.7+/-3.3 years) performed two graded exercise tests on a bicycle ergometer (50 W incline every 3 min) until volitional fatigue in randomized order. During rest and during the last 30 s of each step ventilatory and gas exchange parameters were measured with the CORTEX X1 and the OXYCONgamma. At rest and at each step no significant differences exist for VO2 (F = 0.97) and VCO2 (F = 0.90). The orthogonal regression equation of the VO2-values was VO2(X1) = -75.5 + 1.01 x VO2(Oxy) and the equation of the VCO2-values was VCO2(X1) = 21.7-1.008 x VCO2(Oxy). The VO2-max-values were nearly the same: 3569+/-924 ml/min (Oxy) and 3497+/-993ml/min (X1). Similar findings were made with regard to VCO2max 4117+/-1010 ml/min (Oxy) and 4126+/-1090 ml/ min. (X1). Maximal values for heart rate were 181+/-10 beats/ min (X1) and 180+/-8 beats/min (Oxy) (F=0.21), for maximal power 256+/-64 W (X1) and 257+/-63 (Oxy) (F = 0.0001) and for maximal ventilation 118+/-31 l/min) and 120+/-35 l/min (Oxy) (F = 0.03) with no significant difference. When attached to the motor-driven syringe V(E) was accurately measured up to 288 l/ min. Over a period of 40 min there was no drift observed in F(E)O2 and F(E)CO2. In conclusion, with the CORTEX X1 VO2 and VCO2 can be accurately determined. PMID- 9351693 TI - Relationship between blood lactate response to exercise and endurance performance in competitive female master cyclists. AB - The blood lactate response to graded exercise and its relationship to performance in the field was examined in highly competitive female master cyclists. Thirteen women, age 47.5+/-2.2yr (mean+/-SE), all of whom were United States Cycling Federation (USCF) competitors, underwent laboratory testing for aerobic capacity (VO2max) and lactate threshold (LT), and field testing for performance in 13.5 km and 20 km time-trials. The mean VO2max of the subjects (50.6+/-2.7 ml x kg(-1) x min[-1]) was approximately 10% higher than that previously reported for other age matched female athletes and correlated moderately (r=-0.67) with age. Maximal heart rate was unrelated to age (r=-0.25, p>0.05). Blood lactate concentration (BLC) while time-trialing was significantly higher than that at the LT (2.86+/ 0.17 mmol x l[-1]) for both the 13.5 km (7.59 mmol x l(-1), p < 0.0001), and the 20 km trials (6.99 mmol x l(-1), p<0.002). The LT occurred at a mean power output of 168+/-11.3 W and 66% of VO2max. The VO2 corresponding to a BLC of 4.0 mmol x l(-1) (LT4) was 72% of VO2max. As expected, time-trial performance was highly correlated with VO2max (r=-0.85); however, regression analysis indicated that power output at the lactate threshold was the best laboratory predictor of performance (13.5 km: r2 = 0.83; 20 km: r2 = 0.78). Relative to maximal heart rate, heart rate at LT (88% HRmax) was significantly (p<0.05) lower than time trial heart rate (92-94% HRmax). The cyclists in this study have higher aerobic capacities and maximal heart rates than those previously reported for other age matched female athletes and are able to cycle for extended periods at blood lactate concentrations significantly higher than those at the lactate threshold. Traditional methods of exercise prescription, particularly when using age estimates of maximal heart rate, underestimate training intensities required to be above the LT in female master cyclists. PMID- 9351694 TI - Time course of training-induced changes in maximal exercise of short duration in men and women. AB - The purpose of the present study was to test the hypothesis that gender differences are present in the extent and time course of exercise training induced changes in maximal 10- and 90-s performance test. Thirty-six sedentary subjects (19 women and 17 men) were submitted to 15 weeks of training involving both continuous and interval ergocycle exercise sessions, while 13 other subjects (5 women and 8 men) served as a control group. Maximal power output after 10 s (P10) and 90s (P90) of cycling exercise was measured before and at each 5-week interval of the 15-week training period in both groups. Significant (p < 0.01) training-induced increases in performance were noted after 5 weeks, 10 weeks and 15 weeks of training for P10, and after 5 and 10 weeks for P90 in both genders. P10 and P90 were significantly increased in both genders (about 25% in men and 35% in women) following the 15-week training program and overall absolute increases were not statistically different between men and women. Slight increases (about 5%) in performance tests were observed in control subjects, but only during the first 5-week interval. P10 and P90 of women expressed as a percentage of that of men remained the same throughout the 15-week program. No significant relationship between pre-training values of P10 and their responses to training was found in men and women. In conclusion, results of the present study indicate that women have the same capacity to increase maximal short-term performance in response to training in comparison to men. PMID- 9351695 TI - Influence of acute physical activity and relaxation on state anxiety and blood lactate in untrained college males. AB - The purpose of this investigation was to evaluate the acute effects of physical activity and relaxation on state anxiety and blood lactate. Thirty male Ss performed resistance exercise (N=15) or cycling (N=15) for 50 min at 70% of maximum, while 30 additional male Ss practiced autogenic relaxation (N=15) or rested quietly (N=15) in a sound chamber for 50 min. Assessment of state anxiety and blood lactate was performed before, 5-10 min and 60 min following treatments. The data were analyzed with a repeated measures ANOVA for multifactor experiments, and results indicate a significant group by trial interaction for state anxiety (p<0.0001) and lactate (p < 0.0001). Post-hoc analysis revealed that: 1. lactate increased (p<0.001) immediately following resistance exercise and fell to baseline levels 60 min post exercise; 2. state anxiety was decreased (p < 0.01) at 5-10 min following autogenic relaxation and quiet rest; and 3. a reduction (p < 0.001) in state anxiety was noted at 60 min following cycling. It is concluded that: 1. comparable anxiolytic effects occur following aerobic exercise (cycling), autogenic relaxation and quiet rest, but the effect persists for a longer period of time following aerobic exercise; and 2. accumulation of lactate does not influence state anxiety in normal individuals. PMID- 9351696 TI - VO2max and haemoglobin mass of trained athletes during high intensity training. AB - The correlation between relative haemoglobin mass (Hb mass, g x kg[-1]) and relative maximal oxygen consumption (VO2max, ml x kg(-1) x min[-1]) in 62 trained athletes (33 male runners, 12 male rowers and 17 female rowers) with national and/ or international competitive experience was examined. The correlation between Hb mass and VO2max was highest for the female rowers (n=17, r=0.92, p<0.0001), lower for the male rowers (n = 12, r=0.79, p < 0.005) and lowest for the male runners (n=33, r=0.48, p = 0.005). These results suggest that, within an athletic sample, Hb mass may be used to estimate potential aerobic power. In a second series of experiments, Hb mass was measured before and after three different training programs in sub-sets of the subjects used in the earlier study. Hb mass did not change following 12 weeks of intense rowing training, 4 weeks of heat training (32 degrees C), or 4 weeks of medium-altitude training (1740 m). The corresponding increases in VO2max were 7.8%, no change and 2.1 %, respectively. These results suggest that heat or altitude training does not increase Hb mass in trained athletes. Previous studies that demonstrate increases in total red cell volume following altitude acclimatization used subjects with only modest aerobic power, whereas the present study used trained subjects. It is concluded that trained athletes with erythrocythemic hypervolemia have limited capability to increase further either total red cell volume or Hb mass. PMID- 9351697 TI - Effects of exercise on the macrophage MHC II response to inflammation. AB - The purpose of this investigation was to determine the effects of different doses of exercise on the ability of Propionibacterium acnes (P. acnes) to induce major histocompatibility complex (MHC) II antigen expression on macrophages (M phi's). Pathogen-free male Balb/c mice were exercised on a treadmill moderately (MOD, 15 17 m/min, 5% grade, 30 min/day) or exhaustively (EXH, 15-40m/min, 5% grade, 2 4hr/day) for a period of 7 days during P. acnes-induced inflammation. A control group (CON) consisted of animals exposed to the treadmill environment and handling. Sub-optimal (0.03 mg/g b.wt., i.p.) and optimal (0.08 mg/g b.wt.) doses of P. acnes were used to increase M phi MHC II expression. Animals were sacrified on Day 7 and M phi's were harvested by peritoneal lavage. Direct immunofluorescent staining was performed by incubating peritoneal exudate cells (10[6]) with an FITC-labeled anti-mouse MHC II (I-A[d]) antibody. Basal expression of MHC II was not affected by exercise. There were no significant differences among the groups in the percentage of M phi's expressing MHC II at any dose of P. acnes. However, EXH significantly (p < 0.05) suppressed the expression (mean fluorescent intensity, MFI) of MHC II when compared to MOD (37.1+/-1.95 [mean+/-sem] vs 49.1+/-2.15, p < 0.05) at the suboptimal P. acnes dosage. At the optimal P. acnes dose, both MOD and EXH significantly suppressed (27+/-1.6, 25+/-2.2 and 41.5+/-3.2, for EXH, MOD, and CON, respectively, p<0.0001) P. acnes-induced M phi MHC II MFI. Plasma corticosterone was highly (r= 0.71, p = 0.001) inversely correlated with M phi MHC II expression. However, exercise failed to affect P. acnes-induced production of interferon-gamma. These data suggest that, dependent on the degree of stimulation, exercise can negatively affect M phi expression of MHC II, an effect that may be detrimental to the M phi's ability to present antigen to T lymphocytes. PMID- 9351698 TI - James D. Watson at the Congress of Molecular Medicine. PMID- 9351699 TI - Dinucleotides with a twist: molecular signaling with diadenosine polyphosphates. PMID- 9351700 TI - Molecular databases on the Internet. PMID- 9351701 TI - Genes and politics. PMID- 9351702 TI - Recent progress in studies of neurotrophic factors and their clinical implications. AB - Neurotrophic factors are endogenous soluble proteins that regulate long-term survival and differentiation of neurons of the peripheral and central nervous systems. These factors play an important role in the structural integrity of the nervous system, and therefore are good candidates as therapeutic agents for neurodegenerative diseases. However, recent studies have revealed some unexpected, novel roles of neurotrophic factors. Of particular significance is the discovery of the new functions of brain-derived neurotrophic factor (BDNF) and glia-derived neurotrophic factor (GDNF). Physiological experiments indicate that BDNF may serve as regulatory factors for synaptic transmission as well as for learning and memory. Gene targeting studies demonstrate that GDNF may be essential for development of the enteric nervous system (ENS) and kidney organogenesis. These results not only provide new insights into our understanding of the function of neurotrophic factors but may also have significant implications in the therapeutic usages of neurotrophic factors. PMID- 9351703 TI - Bubonic plague: a molecular genetic case history of the emergence of an infectious disease. AB - Yersinia pestis, the bacterial agent of bubonic plague, is transmitted primarily by fleas and has been responsible for devastating epidemics throughout history. Y. pseudotuberculosis is a food- and water-borne pathogen that causes a much more benign enteric disease in humans. Despite these profoundly different pathogenesis strategies, the two bacteria are very closely related phylogenetically. Thus, identifying the specific genetic differences between them should provide an instructive case study in the evolution of microbial pathogenicity. Some key pathogenesis-related genes of Y. pestis and Y. pseudotuberculosis that have been described to date are compared in this review. Factors that potentiate plague transmission as well as disease are discussed, since dependence on the blood sucking flea for transmission likely fueled the selection of virulent Y. pestis strains able to produce a high-density bacteremia. Retracing the evolutionary steps between these two Yersinia species may ultimately furnish a historical model for the sudden emergence of new human disease agents. PMID- 9351704 TI - Expression of two neuronal markers, growth-associated protein 43 and neuron specific enolase, in rat glial cells. AB - Recent studies have revealed that proteins such as growth-associated protein 43 (GAP-43) and neuron-specific enolase (NSE), believed for many years to be expressed exclusively in neurons, are also present in glial cells under some circumstances. Here we present an overview of these observations. GAP-43 is expressed both in vitro and in vivo transiently in immature rat oligodendroglial cells of the central nervous system, in Schwann cell precursors, and in non myelin-forming Schwann cells of the peripheral nervous system. GAP-43 mRNA is also present in oligodendroglial cells and Schwann cells, indicating that GAP-43 is synthesized in these cells. GAP-43 is also expressed in type 2 astrocytes (stellate-shaped astrocytes) and in some reactive astrocytes but not in type 1 astrocytes (flat protoplasmic astrocytes). These results suggest that GAP-43 plays a more general role in neural plasticity during development of the central and peripheral nervous systems. NSE enzymatic activity and protein and mRNA have been detected in rat cultured oligodendrocytes at levels comparable to those of cultured neurons. NSE expression increases during the differentiation of oligodendrocyte precursors into oligodendrocytes. In vivo, NSE protein is expressed in differentiating oligodendrocytes and is repressed in fully mature adult cells. The upregulation of NSE in differentiating oligodendrocytes coincides with the formation of large amounts of membrane structures and of protoplasmic processes. Similarly, NSE becomes detectable in glial neoplasms and reactive glial cells at the time when these cells undergo morphological changes. The expression of the glycolytic isozyme NSE in these cells, which do not normally contain it, could reflect a response to higher energy demands. This expression may also be related to the neurotrophic and neuroprotective properties demonstrated for this enolase isoform. NSE activity and protein and mRNA have also been found in cultured rat type 1-like astrocytes but at much lower levels than in neurons and oligodendrocytes. Thus GAP-43 and NSE should be used with caution as neuron-specific markers in studies of normal and pathological neural development. PMID- 9351705 TI - Immune-deficient SCID and NOD/SCID mice models as functional assays for studying normal and malignant human hematopoiesis. AB - Many events and requirements of the developmental program of human hematopoietic stem cells have not yet been discovered. A major impediment has been the lack of an appropriate experimental system. At present the conditions for maintaining human stem cells in vitro are not fully known. As a result within a short period the small stem cell pool is lost due to differentiation, making it difficult to examine the correlation between these cells and their function in vivo. Most of our knowledge of hematopoietic stem cells is from animal models in which purified stem cell canididates are assayed based on their functional ability to rescue lethally conditioned recipients. The permanent correction of many genetic disorders of the hematopoietic system requires efficient methods for introducing genes into stem cells in vitro. However, progress has been hindered by the absence of preclinical models that assay the repopulating capacity of primitive human cells. In addition, the development of therapy for malignant diseases also requires assays to identify the target leukemic stem cells based on their ability to initiate the disease. The recent development of methods to transplant or implant both normal and leukemic cells into immune-deficient mice provides the foundation for human stem cell assays. These models assay the repopulating capacity of primitive human cells and provide an important approach to identify and characterize human stem cells, both normal and leukemic. This review focuses on the development of functional assays for normal and leukemic human stem cells and on the new insights that these models are beginning to provide on the organization of the human stem cell hierarchy. PMID- 9351706 TI - Effect of diadenosine polyphosphates on Ca2+ ATPase activity. AB - Diadenosine tri-, tetra-, penta-, and hexaphosphate (Ap3A, Ap4A, Ap5A and Ap6A) have been described as having various effects on vascular tone depending on the number of phosphate groups. This study examined the effect of diadenosine polyphosphates on Ca2+ ATPase activity. The activity of the enzyme was measured spectrophotometrically as the difference in hydrolysis of ATP in the presence and absence of Ca2+ with various concentrations of ATP and diadenosine polyphosphates. The diadenosine polyphosphates increased the activity of the Ca2+ ATPase. The effect tended to be stronger with Ap5A and Ap6A than with Ap3A and Ap4A in the order of potency: Ap3A approximately AP4A < Ap5A approximately AP6A. The stimulatory effect of diadenosine polyphosphates was not competitive with that of ATP, suggesting an allosteric activation of Ca2+ ATPase by diadenosine polyphosphates. This effect may be physiologically relevant for limiting the increase in cytosolic free Ca2+ concentration elicited by diadenosine polyphosphates by receptor activation and modulating Ca2+ ATPase function under resting conditions. PMID- 9351707 TI - Antigen-independent in vitro expansion of T cells does not affect the T cell receptor V beta repertoire. AB - Analysis of the variable chains (V alpha/V beta) of the specific T cell receptor (TCR) of organ-infiltrating T cells may provide further insights into the pathogenesis of many infectious diseases, malignancies, and autoimmune disorders. To determine the TCR V beta repertoire of these small T cell populations antigen independent in vitro expansion is necessary but may select for certain T cell subpopulations. In this study various antigen independent T cell activation protocols were used to stimulate peripheral blood mononuclear cells (PBMC) of six healthy blood donors, and TCR V beta molecules were analyzed by flow cytometry and semiquantitative reverse-transcriptase polymerase chain reaction. In addition, the analysis of in vitro expanded liver-infiltrating T cells and autologous peripheral blood T cells derived from five patients with autoimmune hepatitis but none of six controls revealed a selective overexpression of single TCR V beta molecules in the liver tissue. In contrast to freshly isolated PBMC, no preferential expansion of single TCR V beta families was observed using phytohemagglutinin, anti-CD3 antibodies, or oxidative stress for antigen independent T cell activation. In conclusion, antigen-independent T cell activation offers the chance to analyze small populations of organ-infiltrating T cells without skewing the TCR V beta repertoire. PMID- 9351708 TI - Ultrafast magnetic resonance imaging improves the staging of pancreatic tumors. AB - OBJECTIVE: This prospective study was undertaken to evaluate the accuracy of a noninvasive "all-in-one" staging method in predicting surgical resectability in patients with pancreatic or periampullary tumors. SUMMARY BACKGROUND DATA: Despite progress in imaging techniques, accurate staging and correct prediction of resectability remains one of the chief problems in the management of pancreatic tumors. Staging algorithms designed to separate operable from inoperable patients to save the latter an unnecessary laparotomy are becoming increasingly complex, expensive, time-consuming, invasive, and not without risks for the patient. METHODS: Between August 1996 and February 1997, 58 consecutive patients referred for operation of a pancreatic or periampullary tumor were examined clinically and by 5 staging methods: 1) percutaneous ultrasonography (US); 2) ultrafast magnetic resonance imaging (UMRI); 3) dual-phase helical computed tomography (CT); 4) selective visceral angiography; and 5) endoscopic cholangiopancreatography (ERCP). The assessment of resectability by each procedure was verified by surgical exploration and histologic examination. RESULTS: The study comprised 40 male and 18 female patients with a median age of 63 years. Thirty-five lesions were located in the pancreatic head (60%), 11 in the body (19%), and 1 in the tail of the gland (2%); there were 9 tumors of the ampulla (16%) and 2 of the distal common duct (3%). All five staging methods were completed in 36 patients. For reasons ranging from metallic implants to contrast medium allergy or because investigations already had been performed elsewhere, US was completed in 57 (98%), UMRI in 54 (93%), CT in 49 (84%), angiography in 48 (83%), and ERCP in 49 (84%) of these 58 patients. Signs of unresectability found were vascular involvement in 22 (38%), extrapancreatic tumor spread in 16 (26%), liver metastases in 10 (17%), lymph node involvement in 6 (10%), and peritoneal nodules in only 2 patients (3%). These findings were collated with those of surgical exploration in 47 patients (81 %) and percutaneous biopsy in 5 (9%); such invasive verification was deemed unnecessary and therefore unethical in 6 clearly inoperable patients (10%). In assessing the four main signs of unresectability (extrapancreatic tumor spread, liver metastases, lymph node involvement, and vascular invasion), the overall accuracy of UMRI was 95.7%, 93.5%, 80.4%, as compared to 85.1%, 87.2%, 76.6% for US and 74.4%, 87.2%, 69.2% for CT. In assessing vascular invasion, the sensitivity, specificity, and overall accuracy of angiography were 42.9%, 100%, and 68.8%, respectively. There were 3 complications (12.5%) after 24 resections, 5 in 17 palliative procedures, and none after 6 explorations only. The hospital stay was 14 days after resection, 13 after palliative bypass, and 6 after exploration alone. There was no operative or hospital mortality in these 58 cases. CONCLUSIONS: Although it is by no means 100% accurate, UMRI is equal or even superior to all other staging methods. It probably will replace most of these, because it provides an "all-in-one" investigation avoiding endoscopy, vascular cannulation, allergic reactions, and x radiation. But because even UMRI is not perfect, the final verdict on resectability of a tumor still will depend on surgical exploration in some cases. PMID- 9351709 TI - Long-term survival after retransplantation of the liver. AB - OBJECTIVE: The authors determined the long-term outcome of patients undergoing hepatic retransplantation at their institution. Donor, operative, and recipient factors impacting on outcome as well as parameters of patient resource utilization were examined. SUMMARY BACKGROUND DATA: Hepatic retransplantation provides the only available option for liver transplant recipients in whom an existing graft has failed. However, such patients are known to exhibit patient and graft survival after retransplantation that is inferior to that expected using the same organs in naiive recipients. The critical shortage of donor organs and resultant prolonged patient waiting periods before transplantation prompted the authors to evaluate the results of a liberal policy of retransplantation and to examine the factors contributing to the inferior outcome observed in retransplanted patients. METHODS: A total of 2053 liver transplants were performed at the UCLA Medical Center during a 13-year period from February 1, 1984, to October 1, 1996. A total of 356 retransplants were performed in 299 patients (retransplant rate = 17%). Multivariate regression analysis was performed to identify variables associated with survival. Additionally, a case control comparison was performed between the last 150 retransplanted patients and 150 primarily transplanted patients who were matched for age and United Network of Organ Sharing (UNOS) status. Differences between these groups in donor, operative, and recipient variables were studied for their correlation with patient survival. Days of hospital and intensive care unit stay, and hospital charges incurred during the transplant admissions were compared for retransplanted patients and control patients. RESULTS: Survival of retransplanted patients at 1, 5, and 10 years was 62%, 47%, and 45%, respectively. This survival is significantly less than that seen in patients undergoing primary hepatic transplantation at the authors' center during the same period (83%, 74%, and 68%). A number of variables proved to have a significant impact on outcome including recipient age group, interval to retransplantation, total number of grafts, and recipient UNOS status. Recipient primary diagnosis, cause for retransplantation, and whether the patient was retransplanted before or after June 1, 1992, did not reach statistical significance as factors influencing survival. In the case-control comparison, the authors found that of the more than 25 variables studied, only preoperative ventilator status showed both a significant difference between control patients and retransplanted patients and also was a factor predictive of survival in retransplanted patients. Retransplant patients had significantly longer hospital and intensive care unit stays and accumulated total hospitalization charges more than 170% of those by control patients. CONCLUSIONS: Hepatic retransplantation, although life-saving in almost 50% of patients with a failing liver allograft, is costly and uses scarce donor organs inefficiently. The data presented define patient characteristics and preoperative variables that impact patient outcome and should assist in the rational application of retransplantation. PMID- 9351711 TI - Pancreatoduodenectomy for chronic pancreatitis: anatomic selection criteria and subsequent long-term outcome analysis. AB - OBJECTIVE: The authors sought to provide a framework through outcome analysis to evaluate operations directed toward the intractable abdominal pain of severe chronic pancreatitis centered in the pancreatic head. Pancreatoduodenectomy (PD) was used as an example. SUMMARY BACKGROUND DATA: Head resection for severe chronic pancreatitis is the treatment of choice for a ductal system in the head obliterated by severe disease when associated with intractable abdominal pain. To evaluate the effectiveness of promising head resection substitutes for PD, a framework is necessary to provide a reference standard (i.e., an outcome analysis) of PD. METHODS: Inclusion criteria were severe chronic pancreatitis centered in the pancreatic head, intractable abdominal pain, and a main pancreatic duct obstruction or stricture resulting in absent drainage into the duodenum from the uncinate process and adjacent pancreatic head areas or the entire gland. Since 1986, 57 consecutive cases with these criteria underwent PD (47 head only and 10 total pancreatectomy). Clinical and anatomic predictor variables were derived from the history, imaging studies, and pathologic examination. These variables then were tested for association with the following outcome events gathered during annual follow-up: pain relief, onset of diabetes, body weight maintenance, and peptic ulceration. RESULTS: Operative mortality was zero. In 57 patients with a mean follow-up of 42 months, the 5-year outcome event for survival was 93% and the onset of diabetes was 32%. All new cases of diabetes occurred more than 1 year after resection. In 43 cases > or =1 year postoperative with a mean follow-up of 55 months, all patients indicated significant pain relief and 76% were pain free. Pain relief was more common in patients with diabetes or in those patients with a pancreatic duct disruption. Death was more common in patients with diabetes. Weight maintenance was more common if preoperatively severe ductal changes were not present. Total pancreatectomy was associated with peptic ulceration. CONCLUSIONS: Using selection criteria, the outcome analysis standardized anatomic and clinical variables as to how they were associated with the outcome events (calibrated the effects of the operation with each variable). In these selected patients, PD is safe and significantly relieves pain. Sequelae are from diabetes, provided total pancreatectomy is avoided. PMID- 9351710 TI - Minimally invasive cardiac valve surgery improves patient satisfaction while reducing costs of cardiac valve replacement and repair. AB - OBJECTIVE: This study compares the quality of valve replacement and repair performed through minimally invasive incisions as compared to the standard operation for aortic and mitral valve replacement. SUMMARY BACKGROUND DATA: With the advent of minimally invasive laparoscopic approaches to orthopedic surgery, urology, general surgery, and thoracic surgery, it now is apparent that standard cardiac valve operations can be performed through very small incisions with similar approaches. METHODS: Eighty-four patients underwent minimally invasive aortic (n = 41) and minimally invasive mitral valve repair and replacement (n = 43) between July 1996 and April 1997. Demographics, procedures, operative techniques, and postoperative morbidity and mortality were calculated, and a subset of the first 50 patients was compared to a 50-patient cohort who underwent the same operation through a conventional median sternotomy. Demographics, postoperative morbidity and mortality, patient satisfaction, and charges were compared. RESULTS: Of the 84 patients, there were 2 operative mortalities both in class IV aortic patients from multisystem organ failure. There was no operative mortality in the patients undergoing mitral valve replacement or repair. The operations were carried out with the same accuracy and attention to detail as with the conventional operation. There was minimal postoperative bleeding, cerebral vascular accidents, or other major morbidity. Groin cannulation complications primarily were related to atherosclerotic femoral arteries. A comparison of the minimally invasive to the conventional group, although operative time and ischemia time was higher in minimally invasive group, the requirement for erythrocytes was significantly less, patient satisfaction was significantly greater, and charges were approximately 20% less than those in the conventional group. CONCLUSIONS: Minimally invasive aortic and mitral valve surgery in patients without coronary disease can be done safely and accurately through small incisions. Patient satisfaction is up, return to normality is higher, and requirement for postrehabilitation services is less. In addition, the charges are approximately 20% less. These results serve as a paradigm for the future in terms of valve surgery in the managed care environment. PMID- 9351712 TI - Is hypothermia in the victim of major trauma protective or harmful? A randomized, prospective study. AB - OBJECTIVE: The purpose of this randomized, prospective clinical trial was to determine whether hypothermia during resuscitation is protective or harmful to critically injured trauma patients. SUMMARY BACKGROUND DATA: Hypothermia has both protective and harmful clinical effects. Retrospective studies show higher mortality in patients with hypothermia; however, hypothermia is more common in more severely injured patients, which makes it difficult to determine whether hypothermia contributes to mortality independently of injury severity. There are no randomized, prospective treatment studies to assess hypothermia's impact as an independent variable. METHODS: Fifty-seven hypothermic (T < or = 34.5 C), critically injured patients requiring a pulmonary artery catheter were randomized to a rapid rewarming protocol using continuous arteriovenous rewarming (CAVR) or to a standard rewarming (SR) control group. The primary outcome of interest was first 24-hour blood product and fluid resuscitation requirements. Other comparative analyses included coagulation assays, hemodynamic and oxygen transport measurements, length of stay, and mortality. RESULTS: The two groups were well matched for demographic and injury severity characteristics. CAVR rewarmed significantly faster than did SR (p < 0.01), producing two groups with different amounts of hypothermia exposure. The patients who underwent CAVR required less fluid during resuscitation to the same hemodynamic goals (24,702 mL vs. 32,540 mL, p = 0.05) and were significantly more likely to rewarm (p = 0.002). Only 2 (7%) of 29 patients who underwent CAVR failed to warm to 36 C and both died, whereas 12 (43%) of 28 patients who underwent SR failed to reach 36 C, and all 12 died. Patients who underwent CAVR had significantly less early mortality (p = 0.047). CONCLUSION: Hypothermia increases fluid requirements and independently increases acute mortality after major trauma. PMID- 9351713 TI - Major injury induces increased production of interleukin-10 by cells of the immune system with a negative impact on resistance to infection. AB - OBJECTIVE: The purpose of this study was to compare the production of interleukin 10 (IL-10) by peripheral blood mononuclear cells (PBMC) from injured patients and control subjects to determine the responsible cell types and to relate IL-10 production to the occurrence of sepsis. A mouse model of burn injury was used to confirm the human findings and to assess the importance of IL-10 in the lowered resistance to infection after injury. SUMMARY BACKGROUND DATA: Severe injury is associated with depressed immune responses. Although IL-10 is known to inhibit several aspects of immune reactivity, the role of IL-10 in postinjury immune suppression remains controversial. METHODS: Peripheral blood mononuclear cells from 14 burn and 12 trauma patients and 16 healthy individuals were studied at serial intervals for IL-10 production stimulated by a T-cell mitogen, phytohemagglutinin, and by bacterial lipopolysaccharide. To determine the source of IL-10, CD4+ and CD8+ lymphocyte subsets were obtained by selective depletion of PBMC with antibody-coated magnetic beads and were stimulated by anti-CD3 antibody to induce IL-10 secretion. In addition, IL-10 production by patients' PBMC in the first 10 days after injury was assessed for correlation with subsequent septic events. Anti-CD3-stimulated IL-10 production also was determined for CD4- and CD8-enriched lymphocyte subsets obtained by antibody and complement depletion of splenocytes harvested from groups of burn and sham burn mice at day 10 after injury, the time of maximal susceptibility to a septic challenge, cecal ligation and puncture (CLP). Finally, to test the importance of IL-10 in immune suppression in vivo, groups of burn and sham burn mice were treated with anti-IL-10 monoclonal antibody or control immunoglobulin G (IgG) on days 1 and 3 postinjury and were observed for survival after CLP on day 10. RESULTS: Patients' PBMC produced significantly more IL-10 than did controls' PBMC 7 to 14 days after injury. Patients' CD4+ (T-helper) but not CD8+ (T-cytotoxic) lymphocytes also showed increased IL-10 production versus those of control subjects early after injury. Increased PBMC IL-10 production in the first 10 days postinjury correlated significantly (p < 0.05) with subsequent septic events. Burn mouse CD4-enriched but not CD8-enriched splenocytes produced more IL-10 than did sham burn splenocyte subsets on day 10 after injury. Burn mice treated with anti-IL-10 antibody but not with control IgG had significantly increased survival after CLP. CONCLUSION: Serious injury in humans and in a mouse burn model is followed by increased stimulated production of IL-10 by cells of the immune system. The CD4+ T-helper cells appear to be a major source of IL-10 after injury. In injured patients, increased IL-10 production is correlated with subsequent septic events, and in the burn mouse, IL-10 appears to induce decreased resistance to infection. PMID- 9351714 TI - Implantable left ventricular assist devices: an evolving long-term cardiac replacement therapy. AB - OBJECTIVE: The authors' 8-year experience with both inpatient and outpatient left ventricular assist device (LVAD) support is presented to show the possibilities and limitations of long-term outpatient mechanical circulatory assistance. SUMMARY BACKGROUND DATA: The limitation of suitable cardiac donors has led to the use of LVADs as a temporizing measure for patients awaiting cardiac transplantation. The success of such devices in the short and medium term as a bridge to transplantation has led to their evaluation as a long-term destination therapy for end-stage heart disease. METHODS: Between August 1990 and February 1997, 85 patients with end-stage heart disease underwent insertion of implantable LVADs. Fifty-two patients underwent pneumatic device insertion and 32 patients received a vented electric device. RESULTS: Patients were supported for a mean of 109+/-13 days for an overall survival to transplant (54) or explant (3) of 73%. Nineteen patients were discharged from the hospital on a mean of postoperative day 41+/-4 (range, 17-68) for an outpatient support time of 108+/-30 days (range, 2-466). Of 12 patients supported after postcardiotomy cardiogenic shock, 10 (82%) survived to hospital discharge. Perioperative right ventricular failure was treated in most patients with inotropic agents and inhaled nitric oxide with only six patients requiring right ventricular assist device support. Thromboembolic rate was low (0.016 events/patient-month) despite minimal or no anticoagulation in all cases. CONCLUSIONS: Left ventricular assist device support has evolved to become an outpatient therapy with excellent survival rates and an acceptable morbidity. Accordingly, wearable LVADs should be studied as permanent treatment options for patients who are not transplant candidates. PMID- 9351715 TI - Simultaneous pancreas-kidney transplantation from live donors. AB - OBJECTIVE: In this first report of a clinical series of simultaneous pancreas kidney transplants (SPKs) from live donors, the authors assess donor and recipient outcome as well as the spectrum of surgical and metabolic complications. SUMMARY BACKGROUND DATA: The rationale for live (vs. cadaveric) donation includes an immunologic advantage (better matching, decreased drugs, and fewer rejection episodes) and elimination of waiting time. Only sequential kidney and pancreas or pancreas transplants alone from live donors had been done until the authors' current series. METHODS: Between March 15, 1994, and March 15, 1997, the authors performed 20 SPKs from live donors (6 human leukocyte antigen identical siblings, 14 mismatched relatives [5 parents, 7 siblings, 1 daughter, 1 aunt]). Of the 20 donors, 13 were women, and 7 were men; median age was 43 years (range, 30-58 years). All donors underwent standardized metabolic workup, including oral glucose tolerance tests, determination of hemoglobin A1 c levels, and tests to study insulin secretion and functional insulin secretory reserve. Of the 20 recipients, 12 were women, and 8 were men; median age was 34 years (range, 14-50 years). Management of exocrine pancreatic secretions was with bladder drainage in 17 and duct injection in 3 recipients. Median follow-up was 9 months (range, 1-36 months). RESULTS: Currently, all 20 kidney grafts are functioning. Of the 20 pancreas grafts, 15 are functioning, 3 thrombosed, but 2 of those patients underwent immediate retransplantation from a cadaveric donor, and their grafts currently are functioning. Recipient complications included three anastomotic leaks and three intra-abdominal abscesses. Donor complications included four splenectomies, two peripancreatic fluid collections, one pseudocyst, and one intra-abdominal abscess; two donors underwent reoperation. Three donors had impaired glucose metabolism postdonation. Using tacrolimus and mycophenolate mofetil for mainstay immunosuppression, only 8 of 20 recipients experienced > or =1 rejection episode; only 1 pancreas graft was lost to rejection. Donor and recipient mortality was 0%. CONCLUSION: Simultaneous pancreas-kidney transplants from live donors can be done with no mortality and good graft outcome. With stringent donor criteria, this approach could become another surgical alternative for endocrine replacement therapy in selected patients with uremic type I diabetes. PMID- 9351716 TI - Comparison of open and laparoscopic live donor nephrectomy. AB - OBJECTIVE: This study compares an initial group of patients undergoing laparoscopic live donor nephrectomy to a group of patients undergoing open donor nephrectomy to assess the efficacy, morbidity, and patient recovery after the laparoscopic technique. SUMMARY BACKGROUND DATA: Recent data have shown the technical feasibility of harvesting live renal allografts using a laparoscopic approach. However, comparison of donor recovery, morbidity, and short-term graft function to open donor nephrectomy has not been performed previously. METHODS: An initial series of patients undergoing laparoscopic live donor nephrectomy were compared to historic control subjects undergoing open donor nephrectomy. The groups were matched for age, gender, race, and comorbidity. Graft function, intraoperative variables, and clinical outcome of the two groups were compared. RESULTS: Laparoscopic donor nephrectomy was attempted in 70 patients and completed successfully in 94% of cases. Graft survival was 97% versus 98% (p = 0.6191), and immediate graft function occurred in 97% versus 100% in the laparoscopic and open groups, respectively (p = 0.4961). Blood loss, length of stay, parenteral narcotic requirements, resumption of diet, and return to normal activity were significantly less in the laparoscopic group. Mean warm ischemia time was 3 minutes after laparoscopic harvest. Morbidity was 14% in the laparoscopic group and 35% in the open group. There was no mortality in either group. CONCLUSIONS: Laparoscopic live donor nephrectomy can be performed with morbidity and mortality comparable to open donor nephrectomy, with substantial improvements in patient recovery after the laparoscopic approach. Initial graft survival and function rates are equal to those of open donor nephrectomy, but longer follow-up is necessary to confirm these observations. PMID- 9351717 TI - Prevalence of activating K-ras mutations in the evolutionary stages of neoplasia in intraductal papillary mucinous tumors of the pancreas. AB - OBJECTIVE: The purpose of the study was to determine the prevalence of activating K-ras mutations in the pancreas of patients with intraductal papillary mucinous tumors (IPMT) and to analyze their relation to the degree of site-specific histopathologic abnormality. BACKGROUND: Intraductal papillary mucinous tumors of the pancreas have a biologic behavior that is significantly different from pancreatic ductal adenocarcinoma. Activating K-ras mutations, which may be important events in a multistage process of carcinogenesis, have been reported in IPMT. METHODS: Forty-six different histologic specimens (comprising normal pancreatic ducts, hyperplasia, low-grade dysplasia, high-grade dysplasia carcinoma in situ, and carcinoma) from 16 patients with IPMT and 9 specimens from patients with pancreatic ductal adenocarcinomas were designated by a pathologist. Genomic DNA was extracted from paraffin-embedded tissue sections after microdissection. The K-ras gene was amplified by polymerase chain reaction and subjected to DNA sequencing. RESULTS: The K-ras mutations were detected in at least one specimen in 13 (81.2%) of 16 patients with IPMT. All mutations were found in codon 12. No codon 13 mutations were detected. The relative frequency of K-ras mutations in the different stages of IPMT was 16.7% in normal epithelium and papillary hyperplasia, 28.6% in low-grade dysplasia, and 57.1% in high-grade dysplasia-carcinoma in situ and invasive carcinoma. The K-ras mutations were detected in 6 (66%) of 9 pancreatic ductal adenocarcinomas. CONCLUSIONS: The K ras codon 12 point mutations are as frequent in IPMT as in ductal adenocarcinoma. A stepwise increase in the frequency of codon 12 mutations correlated with the stage of neoplastic evolution to cancer. This finding is consistent with an important role of K-ras gene mutations in the transformation from normal epithelium to invasive carcinoma in the majority of patients with IPMT. PMID- 9351718 TI - Atrial fibrillation after cardiac surgery: a major morbid event? AB - OBJECTIVE: The purpose of the study was to investigate the incidence, predictors, morbidity, and mortality associated with postoperative atrial fibrillation (AF) and its impact on intensive care unit (ICU) and postoperative hospital stay in patients undergoing cardiac surgery in the Department of Veterans Affairs (VA). SUMMARY BACKGROUND DATA: Postoperative AF after open cardiac surgery is rather common. The etiology of this arrhythmia and factors responsible for its genesis are unclear, and its impact on postoperative surgical outcomes remains controversial. The purpose of this special substudy was to elucidate the incidence of postoperative AF and the factors associated with its development, as well as the impact of AF on surgical outcome. METHODS: The study population consisted of 3855 patients who underwent open cardiac surgery between September 1993 and December 1996 at 14 VA Medical Centers. Three hundred twenty-nine additional patients were excluded because of lack of complete data or presence of AF before surgery, and 3794 (98.4%) were male with a mean age of 63.7+/-9.6 years. Operations included coronary artery bypass grafting (CABG) (3126, 81%), CABG + AVR (aortic valve replacement) (228, 5.9%), CABG + MVR (mitral valve replacement) (35, 0.9%), AVR (231, 6%), MVR (41, 1.06%), CABG + others (95, 2.46%), and others (99, 2.5%). The incidence of postoperative AF was 29.6%. Multivariate logistic regression analysis of factors found significant on univariate analysis showed the following predictors of postoperative AF: preoperative patient risk predictors: advancing age (odds ratio [OR] 1.61, 95% confidence interval [CI] 1.48-1.75, p < 0.001), chronic obstructive pulmonary disease (OR 1.37, 95% CI 1.12-1.66, p < 0.001), use of digoxin within 2 weeks before surgery (OR 1.37, 95% CI 1.10-1.70, p < 0.003), low resting pulse rate <80 (OR 1.26, 95% CI 1.06-1.51, p < 0.009), high resting systolic blood pressure >120 (OR 1.19, 95% CI 1.02-1.40, p < 0.026), intraoperative process of care predictors: cardiac venting via right superior pulmonary vein (OR 1.42, 95% CI 1.21-1.67, p < 0.0001), mitral valve repair (OR 2.86, 95% CI 1.72-4.73, p < 0.0001) and replacement (OR 2.33, 95% CI 1.55-3.55, p < 0.0001), no use of topical ice slush (OR 1.29, 95% CI 1.10-1.49, p < 0.0009), and use of inotropic agents for greater than 30 minutes after termination of cardiopulmonary bypass (OR 1.36, 95% CI 1.16-1.59, p < 0.0001). Postoperative median ICU stay (3.6 days AF vs. 2 days no AF, p < 0.001) and hospital stay (10 days AF vs. 7 days no AF, p < 0.001) were higher in AF. Morbid events, hospital mortality, and 6-month mortality were significantly higher in AF (p < 0.001): ICU readmission 13% AF vs. 3.9% no AF, perioperative myocardial infarction 7.41 % AF vs. 3.36% no AF, persistent congestive heart failure 4.57% AF vs. 1.4% no AF, reintubation 10.59% AF vs. 2.47% no AF, stroke 5.26% AF vs. 2.44% no AF, hospital mortality 5.95% AF vs. 2.95% no AF, 6-month mortality 9.36% AF vs. 4.17% no AF. CONCLUSIONS: Atrial fibrillation after cardiac surgery occurs in approximately one third of patients and is associated with an increase in adverse events in all measurable outcomes of care and increases the use of hospital resources and, therefore, the cost of care. Strategies to reduce the incidence of AF after cardiac surgery should favorably affect surgical outcomes and reduce utilization of resources and thus lower cost of care. PMID- 9351719 TI - Ileal pouch-anal canal anastomosis for familial adenomatous polyposis: early and late results. AB - OBJECTIVE: The objective was to review the early and late results of ileal pouch anal anastomosis (IPAA) done for patients with familial adenomatous polyposis (FAP). SUMMARY BACKGROUND DATA: Patients with FAP will have colorectal adenomas develop and die of colorectal cancer if left untreated. Ileal pouch-anal anastomosis removes all disease-bearing mucosa while preserving transanal passage of stools. METHODS: Between 1981 and 1994, 187 patients with FAP, 11 to 59 years of age with a mean follow-up of 60 months (range, 5-170 months) had proctocolectomy and IPAA at Mayo Medical Center in Rochester, Minnesota. All patients had a proximal anal canal mucosal excision and a hand-sewn anastomosis of the pouch to the anal canal at the dentate line. A temporary ileostomy was used in 85% of the patients. RESULTS: No early postoperative deaths occurred, although two patients died later of metastatic colorectal carcinoma present at their initial operation. More important, no patient had a new cancer develop after IPAA. The overall morbidity after operation was 24%, with small bowel obstruction being the most common complication (13%). Patients had four bowel movements/24 hours and good fecal control, which continued during follow-up. CONCLUSIONS: The IPAA eradicates the risk of colorectal cancer in patients with FAP. It can be performed with low mortality, acceptable morbidity, and good functional results over the long term. PMID- 9351720 TI - Inflammation and specialized intestinal metaplasia of cardiac mucosa is a manifestation of gastroesophageal reflux disease. AB - OBJECTIVE: The purpose of the study was to test the hypothesis that cardiac mucosa, carditis, and specialized intestinal metaplasia at an endoscopically normal-appearing cardia are manifestations of gastroesophageal reflux disease. SUMMARY BACKGROUND DATA: In the absence of esophageal mucosal injury, the diagnosis of gastroesophageal reflux disease currently rests on 24-hour pH monitoring. Histologic examination of the esophagus is not useful. The recent identification of specialized intestinal metaplasia at the cardia, along with the observation that it occurs in inflamed cardiac mucosa, led the authors to focus on the type and condition of the mucosa at the gastroesophageal junction and its relation to gastroesophageal reflux disease. METHODS: Three hundred thirty-four consecutive patients with symptoms of foregut disease, no evidence of columnar lined esophagus, and no history of gastric or esophageal surgery were evaluated by 1) endoscopic biopsies above, at, and below the gastroesophageal junction; 2) esophageal motility; and 3) 24-hour esophageal pH monitoring. The patients were divided into groups depending on the histologic presence of cardiac epithelium with and without inflammation or associated intestinal metaplasia. Markers of gastroesophageal reflux disease were compared between groups (i.e., lower esophageal sphincter characteristics, esophageal acid exposure, the presence of endoscopic erosive esophagitis, and hiatal hernia). RESULTS: When cardiac epithelium was found, it was inflamed in 96% of the patients. The presence of cardiac epithelium and carditis was associated with deterioration of lower esophageal sphincter characteristics and increased esophageal acid exposure. Esophagitis occurred more commonly in patients with carditis whose sphincter, on manometry, was structurally defective. Specialized intestinal metaplasia at the cardia was only seen in inflamed cardiac mucosa, and its prevalence increased both with increasing acid exposure and with the presence of esophagitis. CONCLUSION: The findings of cardiac mucosa, carditis, and intestinal metaplasia in an endoscopically normal-appearing gastroesophageal junction are histologic indicators of gastroesophageal reflux disease. These findings may be among the earliest signs of gastroesophageal reflux and contribute to the authors understanding of the pathophysiology of the disease process. PMID- 9351721 TI - Development of a true primary repair for the full spectrum of esophageal atresia. AB - OBJECTIVE: To determine whether or not a true primary repair, without myotomies and with the gastroesophageal junction below the diaphragm, can be accomplished across the esophageal atresia (EA) spectrum. Our hypothesis is that the esophageal anastomosis can withstand significant tension. The consequences, particularly for those patients with a very long gap atresia, were assessed. SUMMARY OF BACKGROUND DATA: Difficulties arise roughly in proportion to the size of the gap between esophageal segments. Reported surgical complications remain frequent, and particularly at the far end of the EA spectrum, not all children are left with a satisfactorily functioning esophagus or esophageal substitute. METHODS: The outcomes of all infants who had a true primary repair of EA from 1976-1997 were determined. Surgically, the methods used to achieve a reliable true primary repair were expanded to accomplish this, even for a very long gap EA. RESULTS: From 1976-97, 70 infants with or without associated tracheoesophageal fistula (TEF) had primary repairs performed with no surgery related deaths and 11% later deaths. No interpositions were performed since 1983. There were no discernible anastomotic leaks and one late recurrent TEF related to the early use of balloon dilation. Ten infants had gaps of 5.0-6.8 cm and, among these, four had gaps of 5.5-6.8 cm that could not be pulled together initially. Traction sutures in the esophageal ends, however, produced sufficient lengthening within 6-10 days for a true primary repair. Very long gap repairs produced more reflux (10 of 10 required a fundoplication versus 24 of 70 overall) and more dilations to relieve strictures. Two infants underwent stricture resection with no recurrence. On follow-up, all patients over 2 years of age were eating well or satisfactorily, and none had a gastrostomy tube. CONCLUSIONS: (1) The esophageal anastomosis can withstand considerable tension and allows a reliable true primary repair for the full EA spectrum. (2) Growth is rapid and traction sutures will produce significant esophageal lengthening within days. (3) With increasing tension, gastroesophageal reflux (GER) and strictures are more common; however, both are treatable. Follow-up reveals the benefits of true primary repair over other solutions. PMID- 9351722 TI - Extracorporeal life support for 100 adult patients with severe respiratory failure. AB - OBJECTIVE: The authors retrospectively reviewed their experience with extracorporeal life support (ECLS) in 100 adult patients with severe respiratory failure (ARF) to define techniques, characterize its efficacy and utilization, and determine predictors of outcome. SUMMARY BACKGROUND DATA: Extracorporeal life support maintains gas exchange during ARF, providing diseased lungs an optimal environment in which to heal. Extracorporeal life support has been successful in the treatment of respiratory failure in infants and children. In 1990, the authors instituted a standardized protocol for treatment of severe ARF in adults, which included ECLS when less invasive methods failed. METHODS: From January 1990 to July 1996, the authors used ECLS for 100 adults with severe acute hypoxemic respiratory failure (n = 94): paO2/FiO2 ratio of 55.7+/-15.9, transpulmonary shunt (Qs/Qt) of 52+/-22%, or acute hypercarbic respiratory failure (n = 6): paCO2 84.0+/-31.5 mmHg, despite and after maximal conventional ventilation. The technique included venovenous percutaneous access, lung "rest," transport on ECLS, minimal anticoagulation, hemofiltration, and optimal systemic oxygen delivery. RESULTS: Overall hospital survival was 54%. The duration of ECLS was 271.9+/-248.6 hours. Primary diagnoses included pneumonia (49 cases, 53% survived), adult respiratory distress syndrome (45 cases, 51 % survived), and airway support (6 cases, 83% survived). Multivariate logistic regression modeling identified the following pre-ECLS variables significant independent predictors of outcome: 1) pre-ECLS days of mechanical ventilation (p = 0.0003), 2) pre-ECLS paO2/FiO2 ratio (p = 0.002), and 3) age (years) (p = 0.005). Modeling of variables during ECLS showed that no mechanical complications were independent predictors of outcome, and the only patient-related complications associated with outcome were the presence of renal failure (p < 0.0001) and significant surgical site bleeding (p = 0.0005). CONCLUSIONS: Extracorporeal life support provides life support for ARF in adults, allowing time for injured lungs to recover. In 100 patients selected for high mortality risk despite and after optimal conventional treatment, 54% survived. Extracorporeal life support is extraordinary but reasonable treatment in severe adult respiratory failure. Predictors of survival exist that may be useful for patient prognostication and design of future prospective studies. PMID- 9351723 TI - A prospective, randomized trial of early enteral feeding after resection of upper gastrointestinal malignancy. AB - OBJECTIVE: The purpose of the study was to determine whether early postoperative enteral feeding with an immune-enhancing formula (IEF) decreases morbidity, mortality, and length of hospital stay in patients with upper gastrointestinal (GI) cancer. SUMMARY BACKGROUND DATA: Early enteral feeding with an IEF has been associated with improved outcome in trauma and critical care patients. Evaluable data documenting reduced complications after major upper GI surgery for malignancy with early enteral feeding are limited. METHODS: Between March 1994 and August 1996, 195 patients with a preoperative diagnosis of esophageal (n = 23), gastric (n = 75), peripancreatic (n = 86), or bile duct (n = 11) cancer underwent resection and were randomized to IEF via jejunostomy tube or control (CNTL). Tube feedings were supplemented with arginine, RNA, and omega-3 fatty acids, begun on postoperative 1, and advanced to a goal of 25 kcal/kg per day. The CNTL involved intravenous crystalloid solutions. Statistical analysis was by t test, chi square, or logistic regression. RESULTS: Patient demographics, nutritional status, and operative factors were similar between the groups. Caloric intake was 61% and 22% of goal for the IEF and CNTL groups, respectively. The IEF group received significantly more protein, carbohydrate, lipids and immune-enhancing nutrients than did the CNTL group. There were no significant differences in the number of minor, major, or infectious wound complications between the groups. There was one bowel necrosis associated with IEF requiring reoperation. Hospital mortality was 2.5% and median length of hospital stay was 11 days, which was not different between the groups. CONCLUSION: Early enteral feeding with an IEF was not beneficial and should not be used in a routine fashion after surgery for upper GI malignancies. PMID- 9351724 TI - Refractory angina pectoris in end-stage coronary artery disease: evolving therapeutic concepts. AB - Refractory angina pectoris in coronary artery disease is defined as the persistence of severe anginal symptoms despite maximal conventional antianginal combination therapy. Further, the option to use an invasive revascularization procedure such as percutaneous coronary balloon angioplasty or aortocoronary bypass grafting must be excluded on the basis of a recent coronary angiogram. This coronary syndrome, which represents end-stage coronary artery disease, is characterized by severe coronary insufficiency but only moderately impaired left ventricular function. Almost all patients demonstrated severe coronary triple vessel disease with diffuse coronary atherosclerosis, had had one or more myocardial infarctions, and had undergone aortocoronary bypass grafting (70% of cases). We present three new approaches with antiischemic properties: long-term intermittent urokinase therapy, transcutaneous and spinal cord electrical nerve stimulation, and transmyocardial laser revascularization. PMID- 9351725 TI - Attenuation over 24 hours of the efficacy of thrombolysis of pulmonary embolism among patients with cancer. AB - The efficacy and safety of thrombolysis in patients with cancer with pulmonary embolism is uncertain. Therefore we studied the effects of thrombolysis in 57 patients with cancer and 254 patients without cancer who were treated in five clinical trials with tissue plasminogen activator or urokinase for pulmonary embolism. Immediately after thrombolysis, the proportion of patients with and without cancer who improved on follow-up angiography (77% vs 73%; p = 0.65) was similar. The angiogrophic reduction in clot burden (1.83 +/- 0.27 vs 1.38 +/- 0.13; p = 0.13) was somewhat greater in patients with cancer than in patients without cancer. Twenty-four hours after initiation of thrombolytic therapy, the proportion of patients who improved on follow-up perfusion scan continued to be similar (72% vs 78%; p = 0.40). However, the extent of reperfusion at 24 hours was less in patients with cancer than in patients without cancer (6% vs 13% reperfusion of lung tissue; p = 0.007). These data suggest that patients with cancer should receive effective anticoagulation in the upper portion of the therapeutic range immediately after thrombolysis. It is possible that such a strategy might preserve initial improvement from thrombolysis and prevent attenuation of benefit during the ensuing 24 hours. PMID- 9351726 TI - Effect of beta-adrenergic blocking agents on mortality rate in patients not revascularized after myocardial infarction: data from a large HMO. AB - We investigated whether patients who do not undergo coronary angiography and therefore any form of revascularization after a myocardial infarction derive greater benefit from chronic beta-blocker therapy than patients who undergo coronary angiography. With multivariate analyses, treatment with beta-blockers was a much stronger predictor of survival in patients who did not undergo coronary angiography (relative risk = 0.38, p = 0.005) than in those patients who did undergo catheterization (p < 0.05 for interaction). Our findings provide direct support for the recommendation by the American College of Cardiology/American Heart Association task force that beta-blocker therapy should be initiated for all infarct survivors who do not undergo revascularization and who have no contraindications. PMID- 9351727 TI - Effects of enalapril during continuous nitrate therapy: analysis of diameter of coronary arteries and platelet cyclic guanosine monophosphate. AB - To investigate the effects of enalapril, an angiotensin-converting enzyme inhibitor, on nitrate tolerance during continuous nitrate therapy, coronary artery diameters and platelet cyclic guanosine monophosphate (cGMP) levels were measured before and 2 minutes after intracoronary injection of nitroglycerin 200 microg in 60 patients with coronary artery disease and were compared among 20 patients treated with nitrates (nitrate group), 20 patients treated with both nitrates and enalapril (enalapril group), and 20 untreated patients (control group). The percent increase in platelet cGMP and coronary dilatation in the nitrate group was significantly less than in the control group, but the percent increase in the enalapril group was significantly greater than that in the nitrate group. These results indicate that enalapril may be helpful as concomitant therapy to maintain the effect of nitrates during continuous nitrate therapy. PMID- 9351729 TI - Prognostic value of clinical markers of reperfusion in patients with acute myocardial infarction treated by thrombolytic therapy. AB - Patients who cannot be reperfused after thrombolytic therapy have a high mortality rate. Noninvasive clinical markers of reperfusion have been widely studied, yet their prognostic significance remains unclear. To assess the prognostic value of commonly used noninvasive clinical markers of early reperfusion we studied 327 patients who received intravenous thrombolytic treatment (1.5 MU streptokinase in 1 hour or 100 mg alteplase in 3 hours) within 6 hours of acute infarction. Successful clinical reperfusion (SCR) was defined as the presence of at least two of the following criteria at 2 hours after thrombolytic treatment: (1) significant relief of pain (a 5-point reduction on a 1 to 10 subjective scale), (2) > or =50% reduction of sum of ST segment elevation, and (3) abrupt initial increase of creatine kinase levels (more than twofold over the upper-normal or baseline elevated values). Clinical variables that were significantly associated by univariate analysis were tested by multivariate analysis to obtain independent predictors of 30-day mortality rate. SCR was present in 210 (64%) patients (group 1), and absent in 117 (36%) patients (group 2). The groups were similar for most baseline characteristics, although group 2 patients were slightly older (mean 60 vs 57 years, p < 0.02). Thirty-day outcomes for group 2 patients compared with group 1 patients were heart failure in 23.1% and 10.5% (p < 0.005), progression to cardiogenic shock in 12.8% and 0.5%, (p < 0.00001), and death in 16.2% and 3.8% (p < 0.0001), respectively. By multivariate analysis the Killip class at admission (p < 0.00001), the absence of SCR (p = 0.017), anterior infarct location (p = 0.021), and age (p = 0.03) were independent predictors of mortality rate, and sex (p = 0.051) had borderline significance. The absence of SCR defined a group of patients with significantly higher mortality rate (odds ratio 4.89, 95% confidence interval 2.07 to 11.57). Three simple noninvasive clinical criteria of successful reperfusion may be used to identify a group of patients with poor prognosis after thrombolytic therapy in whom alternative strategies could be applied. PMID- 9351728 TI - Myoglobin, creatine kinase MB, and cardiac troponin-I to assess reperfusion after thrombolysis for acute myocardial infarction: results from TIMI 10A. AB - BACKGROUND: The availability of a reliable, noninvasive serum marker of reperfusion may permit early identification of patients with occlusion after thrombolysis who might benefit from further interventions. METHODS: We measured myoglobin, creatine kinase MB (CK-MB), and cardiac troponin-I (cTnI) concentrations in sera obtained just before thrombolysis (T0) and 60 minutes later (T60) in 30 patients given TNK-tPA for acute myocardial infarction as part of the Thrombolysis in Myocardial Infarction (TIMI) 10A trial. RESULTS: Angiography at T60 showed reperfusion (TIMI flow grade 2 to 3; n = 19) or occlusion (TIMI flow grade 0 to 1; n = 8). The median serum T60 concentration, the ratio of the T60 and T0 serum concentration, and the slope of increase over a 60-minute period for each serum marker were significantly higher in patients with patent arteries compared with patients with occluded arteries. The areas under the receiver operator characteristics curve for diagnosis of occlusion were 0.96, 0.91, and 0.87 for the T60 concentration of myoglobin, CK-MB and cTnI, respectively. Although the T60 levels of <469 ng/ml for myoglobin, <11.5 ng/ml for CK-MB, and < 1.1 ng/ml for cTnI identified all patients with occlusion, the specificity of myoglobin (94%) was higher than that of CK-MB (61%) and cTnI (67%). Similar results were obtained for the 60-minute ratios and 60-minute slopes for each marker, with indexes for myoglobin having the highest specificity. CONCLUSIONS: In this pilot study, noninvasive diagnosis of occlusion 60 minutes after thrombolysis was achieved with a high degree of sensitivity and specificity with the myoglobin, CK-MB, and cTnI concentrations measured at that time point. These preliminary findings may permit a new strategy for assessment of the success of reperfusion, with triage to rescue angioplasty for patients in whom the 60-minute cardiac marker values or indexes are consistent with occlusion of the infarct-related artery. PMID- 9351731 TI - Accuracy of heart rate-adjusted ST segments in populations with and without posttest referral bias. AB - We compared the accuracy of the ST segment/heart rate (STHR) index and slope to standard criteria (> or =1 mm horizontal/downsloping ST-segment depression at J + 60 msec) in 1358 patients (152 underwent angiography). All exercise tests used the Cornell protocol and computer measurements of maximum ST-segment depression at J + 60 msec. Test accuracy was determined for the entire group with a probability-based method. Thresholds with equal specificity to standard criteria were determined. By using only patients who underwent angiography, neither STHR index nor slope was more accurate than standard criteria (maximum sensitivity: standard criteria, 42%; STHR index, 51%; STHR slope, 40%). However, by using the entire group, both STHR index and slope were more accurate than standard criteria, but only STHR index achieved statistical significance (maximum sensitivity: standard criteria, 31%; STHR index, 60%; STHR slope, 47%). We conclude that heart rate-adjusted ST-segment criteria are more accurate than standard ST-segment criteria. A lack of demonstration of improved accuracy of STHR index and slope only occurs in patients affected by posttest referral bias. PMID- 9351730 TI - Noninvasive prediction of residual blood flow within the risk area during acute myocardial infarction: a multicenter validation study of patients undergoing direct coronary angioplasty. AB - BACKGROUND: In a previous study from a single center, radionuclide measures of collateral flow with technetium 99m sestamibi have been shown to be significantly associated with angiographic residual (antegrade and collateral) flow and independent predictors of final infarct size in acute myocardial infarction. This study examined whether the previously described radionuclide measures of blood flow to the infarct zone were reproducible with different laboratories and imaging systems. METHODS AND RESULTS: Residual flow to the infarct zone was assessed by both invasive and noninvasive methods in 77 patients with first-time myocardial infarction (32 anterior, 45 nonanterior). All patients underwent acute coronary angiography before any intervention within 8 hours of the onset of chest pain (4.0 +/- 1.5 hours; range 1.2 to 7.9 hours). 99mTc sestamibi was injected intravenously before reperfusion therapy, and tomographic imaging was performed 1 to 6 hours after injection. A central core laboratory processed the acquired images from three centers, each with a unique camera and computer system. Three previously published methods based on the severity of the acute perfusion defect were used to measure residual flow to the infarct zone (nadir, severity index, area). Antegrade (Thrombolysis in Myocardial Infarction flow) and collateral flow before direct angioplasty were blindly graded on a four-point scale (0 to 3) from the acute angiogram. The simple sum of the two grades was defined as the angiographic flow index, representing residual flow to the jeopardized zone. All three noninvasive measures of residual flow were highly associated with the angiographic flow index in a linear fashion: severity index (p = 0.0006), area (p = 0.003), and nadir (minimum/maximum counts; p = 0.004). This association was independent of the laboratory where the data were acquired. CONCLUSIONS: Despite different laboratories and camera systems, radionuclide measures of residual flow were highly associated with the angiographic flow index before reperfusion therapy. These results suggest that these measures are applicable on a broader scale for the noninvasive determination of collateral and antegrade flow in acute myocardial infarction. PMID- 9351732 TI - Valvular perforation in left-sided infective endocarditis: a prospective echocardiographic evaluation and clinical outcome. AB - We undertook this study to determine the use of transthoracic and transesophageal echocardiography in detecting valvular perforation and the clinical impact of the latter on the outcome of left-sided infective endocarditis. Transthoracic echocardiography was performed in 58 consecutive patients with infective endocarditis. According to the study protocol, a subgroup of 42 patients also underwent transesophageal echocardiogrophy. At referral, 20 (34%) of 58 patients had echocardiographic evidence of valvular perforation (group A). No valvular perforations were found in the remaining 38 patients (group B). During a follow up period of 27 +/- 16 months, a major complication occurred in 18 of 20 patients in group A and in 11 of 38 patients in group B (p < 0.0001). Univariate analysis indicated previous infective endocarditis, aortic involvement, and New York Heart Association functional class had a predictive value for valvular perforation (p < 0.001). Stepwise regression analysis confirmed aortic valve perforation as the only independent predictive variable for surgery and death. Valvular perforation is a common complication of infective endocarditis and is associated with an adverse outcome. Transthoracic echocardiography can detect or suggest valvular perforation in infective endocarditis, but transesophageal echocardiography better defines this complication and predicts severe heart failure or the need for early surgical management. PMID- 9351733 TI - Echocardiography can detect cloth cover tears in fully covered Starr-Edwards valves: a long-term clinical and echocardiographic study. AB - The incidence of cloth cover tears in fully covered Starr-Edwards valves, as assessed by autopsy or repeat surgery, is approximately 1% per patient-year. However, no echocardiographic study has explored this phenomenon. This study was designed as a one-time observational study and aimed to explore the ability of two-dimensional transthoracic echocardiography to identify cloth cover tears in 35 late survivors with 38 fully covered Starr-Edwards valves who had been operated on 20 to 24 years earlier. The hemodynamic profile, clinical status, and valve-related complications in this highly selected group of late survivors were also studied. Five patients also underwent transesophageal echocardiography. An elongated echogenic mass attached to the prosthetic valve cage and floating downstream was considered indicative of cloth tear. There were 16 patients with aortic valve prostheses, 16 with mitral valve prostheses, and three with double prosthetic valves. In six (17.1%) patients (four with aortic valve prostheses, two with mitral valve prostheses), an echogenic mass suggestive of cloth cover tear was detected, which was confirmed by transesophageal echocardiography in three patients. In two patients the echocardiographic finding was confirmed at surgery. The initial presentation of these six patients was endocarditis, possible embolism, unexplained dyspnea, and weakness in one patient each. Two patients were asymptomatic. There was no evidence of significant prosthetic valve malfunction in any patient. The transvalvular gradients were similar in patients with and without cloth cover tears. Echocardiographic findings highly suggestive of cloth cover tears are not uncommon and can be detected in the third postoperative decade in patients with fully covered Starr-Edwards valves. A prospective study to evaluate the clinical significance of an incidental echocardiographic finding suggestive of cloth cover tears in asymptomatic patients with these valve models is warranted. PMID- 9351734 TI - An agreement approach to predict severe angiographic coronary artery disease with clinical and exercise test data. AB - OBJECTIVE: To demonstrate that an agreement approach to applying equations on the basis of clinical and exercise test variables is an accurate, self-calibrating, and cost-efficient method for predicting severe coronary artery disease in clinical populations. DESIGN: Retrospective analysis of consecutive patients with complete data from exercise testing and coronary angiography referred for evaluation of possible coronary artery disease. After developing an equation in a training set, this equation and two other equations developed by other investigators were validated in a test set. The study was performed at two university-affiliated Veteran's Affairs medical centers. PATIENTS: 1080 consecutive men studied between 1985 and 1995 who had coronary angiography within 3 months of the treadmill test. The population was randomly divided into a training set of 701 patients and a test set of 379 patients. Patients with previous coronary artery bypass surgery, valvular heart disease, marked degrees of resting ST depression, and left bundle branch block were excluded. MEASUREMENTS: Recording of clinical and exercise test data along with visual interpretation of the electrocardiogram recordings on standardized forms and abstraction of visually interpreted angiographic data from clinical catheterization reports. RESULTS: Simple clinical and exercise test variables improved the standard application of exercise-induced ST criteria for predicting severe coronary artery disease. By setting probability thresholds for severe disease of <20% and >40% for the three prediction equations, the agreement approach divided the test set into three groups: low risk (patients with all three equations predicting <21% probability of severe coronary disease), no agreement, and high risk (all three equations with >39% probability) for severe coronary artery disease. Because the patients in the no agreement group would be sent for further testing and would eventually be correctly classified, the sensitivity of the agreement approach was 89% and the specificity was 96%. The agreement approach appeared to be unaffected by disease prevalence, missing data, variable definitions, or even angiographic criteria. CONCLUSIONS: Requiring diagnosis of severe coronary disease to be dependent on agreement between these three equations has made them likely to function in all clinical populations. The agreement approach should be an efficient method for the evaluation of populations with varying prevalence of coronary artery disease, limiting the use of more expensive noninvasive and invasive testing to patients with a higher probability of left main or triple-vessel coronary artery disease. This approach provides a strategy that can be applied by inputting the results of basic clinical assessment into a programmable calculator or a computer to assist the practitioner in deciding when further evaluation is appropriate, thus assuring patients access to subspecialty care. PMID- 9351735 TI - Arterial remodeling after balloon angioplasty of the coronary artery: an intravascular ultrasound study. PICTURE Investigators. PostTreatment IntraCoronary Transluminal Ultrasound Result Evaluation. AB - OBJECTIVE: Before balloon dilation, failure of compensatory enlargement and even arterial shrinkage are frequently observed at the lesion site in response to plaque accumulation. Balloon angioplasty may be regarded as artificial remodeling to enlarge the artery. The prevalence of the different types of arterial wall remodeling after applied stretch by balloon angioplasty is unknown. METHODS AND RESULTS: In 181 patients an intravascular ultrasound study was performed after coronary balloon angioplasty (n = 200 lesions). The vessel area was measured at a proximal and distal reference site and at the lesion site. Subsequently, the relative vessel area [(Vessel area lesion site)/Vessel area reference site) x 100] was calculated. Lesions were classified in three groups on the basis of their relative vessel areas: > or =105%, <105% but >95%, and < or =95%. A relative vessel area > or =105%, indicating enlargement compared with the reference site, was observed in 84 (44%) lesions. A relative vessel area <105% but >95% was observed in 43 (22%) lesions. A relative vessel area < or =95%, indicating "shrinkage" compared with the reference site, was observed in 66 (34%) lesions. CONCLUSIONS: After balloon angioplasty, the vessel area was found to be smaller compared with the reference site in 34% of the lesions. This small vessel area at the lesion site compared with a reference site may be a reflection of insufficient stretch by balloon angioplasty. PMID- 9351737 TI - Cardiovascular complications of malignant carcinoid disease. AB - Carcinoid tumors are endocrinologic malignancies often associated with a characteristic syndrome-the malignant carcinoid syndrome. Cardiovascular manifestations of this rare illness result from unique pathophysiologic characteristics, are associated with poor prognosis, and are difficult to treat medically. The hemodynamic consequences of this disease present unique management problems perioperatively. New pharmacologic and surgical therapies for malignant carcinoids have improved quality of life for patients to the extent that carcinoid heart disease now has more impact on morbidity and mortality rates. Cardiologists may be called on to diagnose and treat this rare cardiac disease. We review, for consulting cardiologists, the pathophysiologic characteristics, cardiovascular manifestations, and management of this disease. PMID- 9351736 TI - Dobutamine stress echocardiography in the detection of coronary artery disease: importance of the pretest likelihood of disease. AB - Although the accuracy of dobutamine stress echocardiography for the detection of coronary artery disease in a high-risk population is known, it has not been well defined for lower risk groups. Two probability groups, high (>75%; n = 199) and intermediate (>10% but < or =75%; n = 118), were studied. Dobutamine stress echocardiography was performed in a standard fashion. Significant coronary artery disease was defined as a >50% luminal diameter stenosis on coronary angiography. The positive predictive accuracy of dobutamine stress echocardiography for the detection of coronary artery disease was greater in the high-probability group (96% vs 86%), as was the sensitivity (89% vs 78%), whereas the negative predictive value was greater in the intermediate-probability group (50% vs 23%), as was the specificity (63% vs 50%). Dobutamine stress echocardiography does have a diagnostic role in the evaluation of patients with an intermediate probability of coronary artery disease. PMID- 9351738 TI - Angioscopic evaluation of angiographically complex coronary lesions. AB - Coronary angioscopy (CA) provides direct visualization of the endoluminal surface of coronary vessels. The usefulness of CA during coronary angioplasty of angiographically complex lesions remains to be established. This study was designed to determine the value of CA to elucidate the underlying substrate of angiographically complex lesions. Forty-seven consecutive patients with angiographically complex lesions were studied with CA before coronary intervention. Mean age of the group was 59 +/- 9 years; six patients were women. Forty (85%) patients had unstable angina. Complex angiographic lesions included coronary occlusions (n = 23) (14 with Thrombolysis in Myocardial Infarction coronary flow grade 0 and nine with flow grade 1), lesions with intraluminal filling defects suggestive of thrombus or ulceration (n = 8), and lesions that were highly eccentric (n = 16). Items analyzed with CA included red thrombus (lining or protruding) and plaque color (yellow, white, or mixed). In all patients, CA visualized the protruding material causing the angiographic appearance. At this site CA detected red thrombus in 34 (72%) patients (14 protruding, 20 lining) and atherosclerotic plaque in 45 (96%) patients. At the site of the angiographically complex lesion, plaque was classified as predominantly yellow in 24 patients, mixed in 12, and white in nine. The incidence of thrombus on CA was higher for occluded vessels (91%) or lesions with intraluminal filling defects or ulceration (87%) than in eccentric lesions (37%) (p < 0.05). However, plaque coloration was not significantly different among these three angiographic subgroups. Initial procedural success (without stent requirement) was lower in lesions showing protruding thrombus on CA (64% vs 91 %, p < 0.05). Thus most angiographically complex lesions contain thrombus. On CA red thrombus was more frequently identified on occluded vessels and lesions with filling defects or ulceration than in eccentric lesions. Yellow or mixed plaques are common in these patients, suggesting lipid-laden plaques as the underlying pathologic substrate of angiographically complex lesions. PMID- 9351739 TI - Effectiveness of tranilast on restenosis after directional coronary atherectomy. AB - Tranilast is an antiallergic drug used widely in Japan that also inhibits the migration and proliferation of vascular smooth muscle cells. This pilot study was undertaken to determine the effectiveness of tranilast on restenosis after successful directional coronary atherectomy. After the procedure, 40 patients (56 lesions, tranilast group) were treated with oral tranilast for 3 months, and 152 patients (188 lesions, control group) did not receive tranilast. Angiographic and clinical variables were compared between the two groups. The minimal lumen diameter was significantly larger in the tranilast group than in the control group at both 3-month (2.08 vs 1.75 mm, p = 0.004) and 6-month follow-up (2.04 vs 1.70 mm, p = 0.003). The diameter stenosis in the tranilast group was smaller than that in the control group both 3 months (28% vs 40%, p = 0.0007) and 6 months (30% vs 43%, p = 0.0001) after the procedure, with a lower restenosis rate (percent diameter stenosis > or =50) in the tranilast group at 3 months (11 % vs 26%, p = 0.03). The number of clinical events over the 12-month period after the procedure was significantly reduced by tranilast administration (p = 0.013). These findings suggest that the oral administration of tranilast strongly prevents restenosis after directional coronary atherectomy. PMID- 9351740 TI - Trends in success rate after percutaneous transluminal coronary angioplasty in men and women with coronary artery disease. AB - Women with coronary artery disease are less likely to undergo percutaneous transluminal coronary angioplasty (PTCA) because of the potential referral bias in favor of men with coronary artery disease in the use of invasive diagnostic procedures and interventions. This difference may represent a sex bias in the delivery of medical care. The apparent sex difference in short-term success of PTCA seen in the early 1980s has not persisted in subsequent studies. The higher in-hospital mortality rate, if any, in women compared with men after PTCA is related more to the severity of their underlying disease rather than sex alone. In addition, women have a better long-term PTCA success rate. PTCA should not be withheld in women who are considered appropriate anatomic candidates for fear of reduced success or increased major complications. PMID- 9351741 TI - Systolic function, readmission rates, and survival among consecutively hospitalized patients with congestive heart failure. AB - We sought to describe the relation between left ventricular systolic function and rates of hospital readmission and survival among consecutively hospitalized patients with congestive heart failure. Medical records were reviewed for these patients at an academic medical center between Jan. 1, 1992, and Dec. 31, 1993. Left ventricular systolic function assessments performed within 6 months before discharge were used to classify left ventricular systolic function. Hospital readmission rates and survival through Dec. 31, 1994, were compared between patients with systolic dysfunction and those with preserved systolic function. Among 412 patients hospitalized with a primary diagnosis of congestive heart failure, 224 had undergone a left ventricular function assessment during the 6 months before hospital discharge. In-hospital mortality and readmission rates were higher among patients without a recent assessment of left ventricular systolic function. Of patients with systolic dysfunction, 55% versus 41% of patients with preserved systolic function were either readmitted or had an emergency room visit within 6 months after discharge (p = 0.06). At 27 months' follow-up, cumulative survival probabilities were 65% for patients with preserved systolic function, 65% for patients with systolic dysfunction, and 60% for patients without a left ventricular systolic function assessment (p = 0.24). Patients without a recent left ventricular systolic function assessment have significantly higher hospital readmission rates than patients with a recent systolic function assessment. Among hospitalized patients, mortality rates are comparable between patients with systolic dysfunction and those with preserved systolic function. However, patients with heart failure with systolic dysfunction may have higher readmission rates. PMID- 9351742 TI - Nitric oxide inhalation reduces pulmonary tidal volume during exercise in severe chronic heart failure. AB - Multiple mechanisms have been proposed to explain the hyperventilation and the limited exercise capacity in congestive heart failure (CHF) including increased intrapulmonary pressures, total pulmonary resistance, and airway abnormalities. We investigated the hypothesis that inhalation of nitric oxide could influence the maximum exercise capacity and excessive ventilatory response to exercise in CHF. Fifteen patients in CHF (mean age 48 +/- 12 years) underwent a control and a nitric oxide inhalation progressive treadmill exercise test with 30 ppm. We determined the maximum oxygen consumptiom (peak VO2), CO2 production (VCO2), minute pulmonary ventilation (VE), respiratory rate, tidal volume (VT), ventilatory equivalent for oxygen (VE/VO2), ventilatory equivalent for carbon dioxide (VE/VCO2), estimated physiologic dead space/tidal volume ratio (VD/VT), VE/VCO2 slope, heart rate, systemic arterial pressure, VE/exercise time slope, and VT/exercise time slope during every incremental exercise. Mean maximum exercise values of heart rate, systolic systemic arterial pressure, diastolic systemic arterial pressure, VD/VT, respiratory rate, peak VO2, VO2/heart rate, VE/CO2, and maximum exercise time were unchanged by inhalation of nitric oxide. There was a strong trend toward reduction of VE/VO2 from 53 +/- 15 to 47 +/- 12 (p = 0.051) and in maximum VE from 58 +/- 21 to 48 +/- 17 L x min(-1) (p = 0.059). Maximum VT decreased from 1639 +/- 556 to 1406 +/- 479 ml (p = 0.04). The VE/VCO2 slope was reduced from 43 +/- 12 to 35 +/- 8 (p = 0.018). Two patients had signs of pulmonary congestion during peak exercise or the recovery period with inhalation of nitric oxide. The VE/exercise time slope and VT/exercise time slope during incremental exercise were reduced by inhalation of nitric oxide, demonstrating a statistically significant minor increase in VE and VT. Inhalation of nitric oxide attenuated the excessive increase in VT response to exercise in CHF. The L-arginine-nitric oxide pathway may be involved in mechanisms contributing to hyperventilation during exercise in CHF. PMID- 9351743 TI - Is left atrial appendage flow a predictor for outcome of cardioversion of nonvalvular atrial fibrillation? A transthroacic and transesophageal echocardiographic study. AB - Accurate echocardiographic parameters for predicting the success of cardioversion or maintenance of sinus rhythm are poorly defined. This prospective transthoracic and transesophageal echocardiographic study was conducted to test the hypothesis that the left atrial appendage flow pattern could be a predictive parameter of the success of cardioversion and maintenance of sinus rhythm in patients with nonvalvular atrial fibrillation. Eighty-two consecutive patients with nonvalvular atrial fibrillation of <6 months' duration underwent transesophageal examination after transthoracic echocardiography. After exclusion of left atrial thrombus, pharmacologic (n = 18) or electrical (n = 64) cardioversion was successful in 75 of 82 patients. In the group that underwent successful cardioversion, maintenance of sinus rhythm (n = 35) or recurrence of arrhythmia (n = 40) was assessed during a 1-year follow-up. During transesophageal examination, five left atrial appendage thrombi were found, spontaneous echo contrast was present in 26 (32%) patients, and mean peak left atrial appendage emptying velocity was 35 +/- 18 cm/sec. Peak left atrial appendage emptying velocity was found to be statistically related to parameters of left ventricular and left atrial function but not to long-term maintenance of sinus rhythm. No other echocardiographic parameter was identified as a predictor for either the success of cardioversion or the maintenance of sinus rhythm at follow-up. In patients with nonvalvular atrial fibrillation of recent onset, peak left atrial appendage emptying velocity appears to be a complex parameter depending on left atrial and left ventricular function but that does not predict either the success rate of cardioversion or long-term maintenance of sinus rhythm after successful cardioversion. PMID- 9351745 TI - Blood pressure and cardiovascular morbidity and mortality rates in the elderly. AB - The influence of blood pressure on the development of cardiovascular disease and mortality rate beyond 75 years of age has been uncertain. A reported inverse relation to mortality rate noted in the very old could reflect poor cardiovascular health. This report examines the impact of blood pressure on cardiovascular morbidity and mortality rates and all-cause mortality rate in those aged 75 to 94 years. For this analysis the data were the Framingham study subjects found free of cardiovascular disease and a second sample of those with cardiovascular disease present on the biennial examinations. Investigation of the relation of systolic and diastolic blood pressures to all causes and cardiovascular morbidity and mortality rates within each 2-year interval for those aged 75 to 94 years was carried out. Over the period of 38 years of follow up there were increasing cardiovascular morbidity and mortality rates with increasing blood pressure levels for both men and women in the sample free of cardiovascular disease. In those with cardiovascular disease at the biennial examination there was a distinct U-shaped curve of cardiovascular mortality rate in relation to systolic blood pressure in men with a substantial increase in mortality rate below systolic pressures of 120 mm Hg for both men and women. Excess mortality rate reported in elderly persons with low blood pressure appears to be a result of poor cardiovascular health and not from the low pressure itself. The excess mortality rate reported for low blood pressure levels in persons older than 75 years derives from the inclusion of the substantial proportion of this age group who have cardiovascular disease. PMID- 9351744 TI - Circadian variation of sustained ventricular tachycardia in patients subject to standard adrenergic blockade. AB - Morning peaks in the circadian variation of sustained ventricular tachycardia (VT) may reflect the contribution of sympathetic activation to onset of VT. We hypothesized that adrenergic blockade would eliminate this morning peak. Fifty four patients using a defibrillator had 1114 time-stamped episodes of VT requiring therapy with a device: 1012 episodes with and 102 episodes without antiadrenergic medications. Nine patients had episodes both with and without antiadrenergic medication and were examined separately. In patients taking antiadrenergic agents, data fitted to a harmonic regression model revealed a morning peak at 9:00 AM (R2= 0.542; p < 0.05), with a secondary peak at 4 PM. Those not receiving antiadrenergic therapy had a similar morning peak. Antiadrenergic agents as used in standard clinical practice do not prevent circadian variation in onset of VT. This variation may be mediated by systems other than adrenergic receptor-linked activation or may reflect inadequacy of adrenergic blockade in standard clinical dosing. PMID- 9351746 TI - Effects of hormone-replacement therapy on hemostatic factors, lipid factors, and endothelial function in women undergoing surgical menopause: implications for prevention of atherosclerosis. AB - Women with premature menopause are at high risk for vascular compications associated with thrombogenesis and atherogenesis. The use of hormone-replacement therapy (HRT), however, may protect against these complications. Hemostatic abnormalities and endothelial function are closely related to the processes of thrombogenesis and atherogenesis. The purpose of the study was to evaluate the effects of premature menopause on markers of hemostasis, platelet function, and endothelial function and the effects of starting HRT. This is a prospective longitudinal study of premenopausal women undergoing surgical menopause in whom estrogen HRT is started. We measured sequential changes in plasma levels of the hemostatic factors (fibrinogen, fibrin D-dimer, and plasminogen activiator inhibitor [PAI]), markers of platelet function (soluble leukocyte adhesion molecule P-selectin) and endothelial function (von Willebrand factor [vWf], soluble thrombomodulin [sTM], and tissue plasminogen activator [TPA]), and serum lipid levels, including lipoprotein A. Twenty-seven premenopausal women (mean age 43.6 +/- 6.5 years) undergoing hysterectomy and bilateral salpingo-oophrectomy were studied. In the postsurgical menopausal state (visit 2), there was a significant elevation in sTM levels (paired Wilcoxon test, p = 0.008). There was also a trend toward higher median soluble P-selectin, PAI, and mean TPA levels and lower vWf levels. After 6 weeks of HRT (visit 3), there was a significant reduction in mean vWf (paired Wilcoxon test, p = 0.0026), sTM (p = 0.039), and TPA levels (p = 0.02) compared with premenopausal levels. There were no significant changes in plasma fibrinogen, fibrin D-dimer, and PAI levels at visit 2 or visit 3 compared with premenopausal levels. There was a significant increase in serum lipoprotein A (paired Wilcoxon test, p = 0.008), cholesterol, and triglyceride levels after surgical menopause (paired t test, p < 0.01). Lipoprotein A and cholesterol levels after HRT (visit 3) were not significantly different from prehysterectomy levels, although triglyceride levels were increased further. HRT results in a significant reduction in vWf, sTM, and TPA levels, suggesting beneficial effects on endothelial function and atherogenesis. Although there was a significant increase in serum lipoprotein A and cholesterol levels after surgical menopause, lipoprotein A and cholesterol levels after HRT were not significantly different from presurgery levels. These observations are consistent with the beneficial effects of HRT in cardiovascular hemodynamics and cardiovascular disease. PMID- 9351747 TI - A novel prognostic scoring system to predict late outcome after percutaneous balloon valvotomy in patients with severe mitral stenosis. AB - We developed a prognostic scoring system to predict the outcome of follow-up after balloon mitral valvotomy. The system incorporates seven variables before valvotomy: age, New York Heart Association class, fluoroscopic calcification, echocardiographic score, cardiac rhythm, mitral regurgitation, and mitral valve area. Each variable was coded with either 0 or 1 and a total score was between 0 and 7. The study included 150 patients with a mean follow-up of 33 +/- 24 months. In patients with scores of 0-1, 2-3, 4-5, and 6-7, the estimated cardiac event free survival rate was 97%, 94%, 86%, and 68%, respectively, at 1 year; 95%, 88%, 74%, and 47%, respectively, at 3 years; and 92%, 82%, 61%, and 30%, respectively, 5 years after valvotomy (p = 0.0001). The hazard risk ratio for cardiac events was 1.7 times greater for every step up of the score (p = 0.0001). Our scoring system provides a simple but effective method to predict late outcome of balloon mitral valvotomy. PMID- 9351748 TI - Increased accumulation of acidic fibroblast growth factor in left ventricular myocytes of patients with idiopathic cardiomyopathy. AB - To clarify the pathophysiologic role of fibroblast growth factors in idiopathic cardiomyopathy, we evaluated endomyocardial biopsy specimens obtained from 24 patients (nine with hypertrophic cardiomyopathy [HCM], 12 with dilated cardiomyopathy [DCM], and three with hypertensive hypertrophy) and six controls. All the specimens were stained for acidic fibroblast growth factor (aFGF) and basic FGF (bFGF) with immunohistochemistry. In situ hybridization was carried out for detection of aFGF mRNA. The average diameter of the myocytes, the percent area of interstitial fibrosis, and capillary vessel density were assessed in each biopsy specimen with morphometric methods. Positive staining of aFGF was observed in the myocytes of the biopsy specimens taken from 15 of 21 (71%) patients with cardiomyopathy (six of nine HCM, nine of 12 DCM) and all hypertensive hypertrophy patients but in none of the controls (p < 0.01). The average diameter of the myocytes was significantly larger in the patients with positive aFGF staining than in those with negative staining (23.1 +/- 1.5 versus 18.3 +/- 1.2 microm, p < 0.05). The percent area of fibrosis and the density of capillaries did not differ between the two groups. Intense expression of aFGF mRNA was observed in the myocytes from the patients with positive aFGF protein. In conclusion, the expression of FGF was significantly increased in myocytes obtained from the left ventricle of patients with cardiomyopathy. Acidic FGF may contribute to the hypertrophy of myocytes as the repair response to myocardial injury in patients with idiopathic cardiomyopathy. PMID- 9351749 TI - Progesterone metabolite allopregnanolone in women with premenstrual syndrome. AB - OBJECTIVE: To evaluate the anxiolytic 3alpha-5alpha-reduced progesterone metabolite allopregnanolone in the luteal phase of the menstrual cycle in women with premenstrual syndrome (PMS) and controls. METHODS: Thirty-five women with prospectively documented PMS and 36 controls were evaluated. Serum progesterone and allopregnanolone levels were measured on days 19 and 26 of the cycle as determined by urinary LH detection kits. Analysis of variance and Student t tests were used to analyze the data. RESULTS: Allopregnanolone levels were significantly lower on day 26 in the PMS group than in controls (3.6 +/- 0.8 versus 7.5 +/- 1.3 ng/mL; P < .04). Significant differences in the ratio of the metabolite to progesterone also were noted, with a smaller ratio in the PMS subjects (0.9 +/- 0.3 versus 3.2 +/- 1.3 ng/mL; P < .05). There were no significant differences between the PMS and control groups with respect to serum progesterone levels. CONCLUSION: Subjects with PMS manifested lower levels of the anxiolytic metabolite allopregnanolone in the luteal phase when compared with controls. Diminished concentrations of allopregnanolone in women with PMS may lead to an inability to enhance gamma aminobutyric acid-mediated inhibition during states of altered central nervous system excitability, such as ovulation or physiologic or psychological stress. The lowered metabolite levels could contribute to the genesis of various mood symptoms of the disorder, such as anxiety, tension, and depression. PMID- 9351750 TI - A randomized controlled trial to evaluate the use of the endocervical brush after endocervical curettage. AB - OBJECTIVE: To determine if using the endocervical brush after curetting the endocervix will increase the yield of endocervical tissue retrieved for an endocervical curettage (ECC) specimen. METHODS: Between March 1, 1995, and June 30, 1996, we recruited for participation patients with abnormal Papanicolaou smears referred for colposcopy. Exclusion criteria were pregnancy, previous hysterectomy, and a history of diethylstilbestrol exposure. Colposcopy and biopsies were performed by residents under direct supervision of the attending staff. Endocervical curettages were performed using the Kevorkian endocervical curette. The subjects were then assigned randomly to one of two ways of collecting the ECC tissue: with either a curette or an endocervical brush. Specimens were reviewed by pathologists, who were blinded to the method of ECC collection. RESULTS: During the study period, 124 patients agreed to participate; 62 were assigned to the control group, and 62 to the study group. Six subjects had missing data, leaving 118 patients available for analysis. In the control group, six of the 58 ECC samples obtained contained insufficient endocervical tissue for pathologic diagnosis. None of the 60 samples from the endocervical brush group was insufficient. The difference between the two groups was statistically significant (P = .01). CONCLUSION: The addition of the endocervical brush to endocervical tissue sampling at colposcopy in the study decreased the number of insufficient samples. The endocervical brush method of collection of an ECC specimen from the canal after the Kevorkian curette is used is a valuable addition to this diagnostic tool. We recommend its use in obtaining an ECC specimen. PMID- 9351752 TI - Factors affecting detrusor contraction strength during voiding in women. AB - OBJECTIVE: To compare voiding mechanisms in continent and stress incontinent women and to assess the effects of aging, childbirth, menopausal status, and anterior vaginal wall relaxation on detrusor contraction strength during voiding. METHODS: Thirty-eight asymptomatic female volunteers underwent a thorough evaluation including multichannel urodynamic testing and instrumented pressure flow voiding studies. The voiding mechanisms and detrusor contraction strengths, available in 30 women, were compared with those of 70 women evaluated previously with objective evidence of genuine stress urinary incontinence. The effect of age, parity, bladder neck mobility, and anterior vaginal wall relaxation on maximum detrusor pressure was assessed using chi2 and linear regression analyses. Detrusor pressures in premenopausal and postmenopausal women and continent and stress incontinent women were also compared. RESULTS: Four types of voiding mechanisms were identified. All 30 of the continent women voided with a detrusor contraction, compared with 59 (84%) of genuine stress incontinent subjects. The mean +/- standard deviation [SD]) detrusor contraction was significantly stronger in continent women than incontinent women (20.3 +/- 14.2 cm H2O compared with 12.3 +/- 11.0 cm H2O; P < .01). In continent and incontinent subjects, maximum detrusor pressure did not correlate significantly with increasing age, parity, bladder neck mobility, or degree of anterior vaginal wall relaxation. Premenopausal women had significantly higher mean (+/- SD) maximum detrusor pressures than postmenopausal women (16.3 +/- 13.0 cm H2O compared with 11.5 +/- 11.0 cm H2O; P < .01). CONCLUSION: Women with genuine stress urinary incontinence may be more likely to void with a weak or absent detrusor contraction than continent women. Menopausal status was the only factor identified that significantly affected maximum detrusor pressure during voiding. PMID- 9351751 TI - Does prolonged high-impact activity contribute to later urinary incontinence? A retrospective cohort study of female Olympians. AB - OBJECTIVE: To determine whether women engaged in strenuous, provocative exercise are more likely to be incontinent in future life than similarly fit women who participated in less provocative exercise. METHODS: In this retrospective cohort study, female American Olympians who competed in swimming (low-impact group) and in gymnastics and track and field (high-impact group) between 1960 and 1976 completed a structured questionnaire. Primary outcome measures included the prevalence of the symptoms of stress and urge incontinence. Statistical analyses of results included chi2, Fisher exact test, two-tailed t tests, Wilcoxon rank sum test, and stepwise multiple logistic regression. P < .05 was considered significant. RESULTS: One hundred four women responded (response rate 51.2%). High-impact athletes were older (46.2 compared with 42.4 years) and were more likely to report incontinence when they were doing their sport as Olympians (35.8% compared with 4.5%) than low-impact athletes; low-impact athletes were more likely to be parous (83.3% compared with 60.7%). There was no difference in the prevalence of the symptom of stress incontinence between the high- versus low impact groups: any incontinence, 41.1% compared with 50%; daily or weekly incontinence, 10.7% compared with 8.3%; and incontinence that bothered them moderately or greatly, 10.7% compared with 4.2%. With our sample size, this study had 80% power to detect a fourfold difference in daily or weekly incontinence between groups, but only a 30% power to detect a twofold difference, given a baseline prevalence of 10%. When age, body mass index (BMI), parity, Olympic sport group, and incontinence during Olympic sport were entered into stepwise logistic regression analyses, only BMI was significantly associated with regular stress or urge incontinence symptoms. CONCLUSION: Participation in regular, strenuous, high-impact activity when younger did not predispose women to a markedly higher rate of clinically significant urinary incontinence in later life. PMID- 9351753 TI - EndothelinA receptors in human uterine leiomyomas. AB - OBJECTIVE: To determine if there are endothelin receptors on human uterine leiomyomas. METHODS: Samples of leiomyomas from eight patients were analyzed for [iodine (I)-125]endothelin-1 binding. Several subtype-selective ligands were used to determine the endothelin receptor population. RESULTS: Binding of [125I]endothelin-1 to uterine leiomyoma membranes was specific and saturable, with a mean +/- dissociation constant 85.5 +/- 8.4 pM. Competition binding studies showed that the order of potency was endothelin-1 > endothelin-3, which was consistent with the presence of the endothelinA receptor subtype. Binding of [125I]endothelin-1 was displaced by an endothelinA-selective antagonist, but not by sarafotoxin 6c, an endothelinB-selective agonist. An endothelins-selective ligand was not specifically bound to leiomyoma. CONCLUSION: These results indicate that only endothelinA receptors are present in human uterine leiomyomas. We speculate that endothelin-1 may act through these endothelinA receptors to influence the development or regulation of hypertrophy and proliferation of the human myometrium during pregnancy and in uterine disorders like leiomyomas. PMID- 9351754 TI - Postoperative pain relief following laparoscopic tubal sterilization with silastic bands. AB - OBJECTIVE: To evaluate postoperative pain relief of intramuscular ketorolac, topical bupivacaine, and placebo in patients undergoing laparoscopic tubal sterilization with silastic bands. METHODS: One hundred five women undergoing laparoscopic tubal sterilization with silastic bands were randomized to one of three groups: one received intramuscular ketorolac and topical placebo applied to the fallopian tubes, the second received intramuscular placebo and topical bupivacaine, and the third received intramuscular placebo and topical placebo. Surgical procedures, anesthesia, and recovery were conducted with standardized protocols. Postoperative pain perception was graded using the modified McGill pain intensity scale at 30 minutes postoperatively, at discharge from the recovery room, and the next morning by telephone interview. Other measured variables included postoperative vomiting, additional analgesia requirement, and length of time spent in the recovery room. RESULTS: Only topical bupivacaine was found to decrease postoperative pain scores significantly over those with placebo, at 30 minutes postoperatively (median score 2 compared with 4, P = .002) and at discharge from the recovery room (median score 2 compared with 3, P = .03). There was no significant decrease in pain scores with intramuscular ketorolac compared with placebo. No differences in pain scores were found between the three groups at the next morning phone call. There were no significant differences between the three groups with respect to requirements for supplemental pain medications in the recovery room, incidence of postoperative vomiting, or length of time spent in the recovery room. CONCLUSION: Topical bupivacaine decreases postoperative pain scores significantly compared with placebo in women undergoing laparoscopic tubal sterilization with silastic bands. PMID- 9351755 TI - Vaginal misoprostol compared with oral misoprostol in termination of second trimester pregnancy. AB - OBJECTIVE: To compare the efficacy of vaginal with oral misoprostol in termination of second-trimester pregnancy after pretreatment with mifepristone. METHODS: Women requesting termination of second-trimester pregnancy were randomized into two groups. Thirty-six to 48 hours after oral administration of 200 mg of mifepristone, women were given either oral or vaginal misoprostol 200 microg every 3 hours for a maximum of five doses in the first 24 hours. Women receiving oral misoprostol also were given a vaginal placebo (vitamin B6), whereas those receiving vaginal misoprostol were given an oral placebo. If they failed to abort, a second course was given by the same route. RESULTS: The median induction-abortion interval in the vaginal group (9 hours) was significantly shorter than that in the oral group (13 hours). The percentage of women aborting within 24 hours in the vaginal group (90%) was significantly higher than that in the oral group (69%). The median amount of misoprostol used in the vaginal group (600 microg) also was significantly less than that in the oral group (1000 microg). There was no significant difference in the incidence of side effects between the two groups except for fatigue and breast tenderness, which were more common in the oral group. Seventy-six percent of the women preferred the oral route, and 24.5% of the women preferred the vaginal route. CONCLUSION: Vaginal misoprostol is more effective than oral misoprostol in termination of second trimester pregnancy after pretreatment with mifepristone, but more women preferred the oral route. PMID- 9351756 TI - Cervicovaginal human immunodeficiency virus secretion and plasma viral load in human immunodeficiency virus-seropositive women. AB - OBJECTIVE: To evaluate human immunodeficiency virus (HIV)-1 RNA burden in paired plasma and cervicovaginal lavage specimens and to assess the relation of plasma HIV-1 RNA level, CD4 cell count, and antiretroviral therapy with cervicovaginal HIV-1 viral load. METHODS: Paired blood and cervicovaginal lavage specimens were collected from 72 HIV-infected women. Quantitation of HIV-1 RNA from plasma and cervicovaginal lavage specimens was performed by using the nucleic acid sequence based amplification assay. Analyses examined relations between cervicovaginal HIV 1 RNA and plasma HIV-1 RNA level, CD4 count, and antiretroviral therapy. RESULTS: Plasma HIV-1 RNA was detectable in 61 of 72 women (85%), with copy numbers ranging from 330 to 1,600,000 copies/mL. Twenty-eight of 72 (39%) had detectable HIV-1 RNA in cervicovaginal lavage specimens, ranging from 320 to 440,000 copies/mL. The cervicovaginal lavage HIV-1 RNA level was detectable in 9%, 29%, 52%, and 53% of the women with plasma HIV-1 RNA of less than 400, 400-9999, 10,000-100,000, and more than 100,000 copies, respectively (P = .043). Among women with CD4 counts of less than 200, 200-500, and greater than 500/mm3, cervicovaginal lavage HIV-1 RNA was detected in 67%, 32%, and 25% of subjects, respectively (P = .018). Among women receiving antiretroviral therapy, cervicovaginal lavage revealed HIV-1 RNA in 67%, 31%, and 25% with CD4 cell counts of less than 200, 200-500, and more than 500/mm3, respectively (P = .042). CONCLUSION: The presence of HIV-1 RNA in cervicovaginal lavage correlates significantly with the level of HIV-1 RNA in plasma and negatively with CD4 cell count. PMID- 9351757 TI - Acetowhitening of the cervix and vulva as a predictor of subclinical human papillomavirus infection: sensitivity and specificity in a population-based study. AB - OBJECTIVE: To evaluate acetowhite changes of the cervix and vulva as a predictor of human papillomavirus (HPV) infection. METHODS: In this population-based study all women aged 19, 21, 23, and 25 years and registered as living in a primary health care area within the city of Umea, Sweden were eligible for inclusion. Each participant underwent a gynecologic examination with sampling of epithelial cells for HPV-DNA detection and Papanicolaou smear. Colposcopy was performed 5 minutes after application of 5% acetic acid. A two-step polymerase chain reaction (PCR) technique was employed for HPV-DNA detection. RESULTS: Colposcopy and sampling of epithelial cells could be performed in 535 women. The sensitivity of detection of HPV infection by the acetowhitening of the cervix was 22% (95% confidence interval [CI] 18%, 26%). The specificity of detection of HPV infection by the acetowhitening of the cervix was 90% (95% CI 87%, 93%). The sensitivity of detection of HPV infection by cytology was 13% (95% CI 10%, 16%), and the specificity was 99% (95% CI 98%, 100%). The combination of acetowhitening and cytology did not improve the diagnostic value. CONCLUSION: Acetowhitening of the cervix and vulva has low sensitivity as a predictor of HPV infections as determined by PCR. PMID- 9351758 TI - Human papillomavirus type 16 and risk of preinvasive and invasive vulvar cancer: results from a seroepidemiological case-control study. AB - OBJECTIVE: To examine whether human papillomavirus (HPV) type 16 is involved in the etiology of vulvar carcinomas. METHODS: We studied 142 histologically confirmed cases of vulvar intraepithelial neoplasia (VIN) grade 3 and invasive vulvar cancer and 126 community controls. In addition to a detailed questionnaire through which we obtained information on putative risk factors for vulvar cancer, blood samples were collected from participating subjects and tested for the presence of antibodies to HPV-16 virus-like particles. Data were analyzed by logistic regression. RESULTS: Subjects positive for HPV-16 antibodies were at a 5.3-fold increased risk of vulvar neoplasia (95% confidence interval [CI] 2.5, 11.1), and subjects with high antibody levels were at a 20-fold increased risk of disease (95% CI 5.4, 76.7). A stronger association between HPV-16 seropositivity and disease was observed for VIN grade 3 (odds ratio [OR] 13.4; 95% CI 3.9, 46.5) than for invasive disease (OR 2.9; 95% CI 0.94, 8.7), and for invasive tumors, there was a suggestion that the association was stronger for women diagnosed with squamous carcinoma of basaloid and/or warty types (OR 3.8; 95% CI 0.76, 18.9) than for those diagnosed with keratinizing squamous cell carcinomas (OR 1.6; 95% CI 0.35, 7.4). Number of sexual partners and herpes simplex virus type 2 seropositivity remained as independent risk factors for vulvar neoplasia after control for confounding by HPV-16. The risk associated with HPV-16 seropositivity was higher among smokers (OR 8.5; 95% CI 3.8, 19) than among nonsmokers (OR 3.4; 95% CI 0.85, 13). CONCLUSION: Our results confirm that HPV is associated with vulvar carcinomas. Findings also suggest the possibility that other sexually transmitted agents might be involved in the etiology of some vulvar tumors and that smoking may be an important cofactor involved in the etiology of HPV-related vulvar tumors. Evaluation of the role of HPV types other than HPV-16 in the etiology of vulvar cancer is needed, and additional efforts aimed at further elucidating the role of smoking and other cofactors in this disease process are warranted. PMID- 9351759 TI - Order of endocervical and ectocervical cytologic sampling and the quality of the Papanicolaou smear. AB - OBJECTIVE: To determine whether the order of cell collection, endocervical or ectocervical cells first, has an effect on the quality of the Papanicolaou smear. METHODS: One thousand smears were obtained using an Ayre spatula and an endocervical brush. In 500 cases the endocervical brush was used first, and in 500 cases the spatula was used first. All Papanicolaou smears were collected by resident physicians in our university hospital gynecologic clinics. A smear was considered limited for interpretation for the following reasons: 1) lack of endocervical component, 2) obscured by blood, 3) obscured by inflammation, 4) drying artifact, and 5) too thick. RESULTS: The brush-first group had 405 (81%) adequate smears compared with 410 (82%) adequate smears in the spatula-first group. More smears were obscured by blood when the brush was used first (22 or 4.4% compared with three or 0.6%, P < .001). No endocervical component (ie, metaplastic cells, endocervical cells, or mucus) was found in 29 (5.8%) smears from the brush-first group compared with 45 (9.0%) of the spatula-first group, an insignificant difference. More squamous intraepithelial lesions were found when the spatula was used first (55 or 11% compared with 35 or 7.0%, P < .05). CONCLUSION: The quality of the Papanicolaou smear can be improved by using the Ayre spatula first followed by the endocervical brush. Fewer smears will be obscured by blood, which could result in more squamous intraepithelial lesions being detected. PMID- 9351760 TI - Risk of residual invasive disease in women with microinvasive squamous cancer in a conization specimen. AB - OBJECTIVE: To quantify the risk of residual invasion when cervical conization reveals microinvasive squamous carcinoma and to determine whether any factors affect this risk. METHODS: We reviewed the charts and histopathology slides of 87 women who underwent a conization that contained microinvasive squamous carcinoma, followed by either a repeat conization or hysterectomy. Depth of invasion, number of invasive foci, and status of the internal margin and post-conization endocervical curettage (ECC) were assessed. The findings were correlated with the presence of residual invasion. RESULTS: Significant predictors of residual invasion included status of the internal margin (residual invasion present in 22% of women with an involved margin versus 3% with a negative margin; P < .03) and the combined status of the internal margin and post-conization ECC (residual invasion in 4% of patients if both negative, 13% if one positive, and 33% if both positive; P < .015). Depth of invasion and number of invasive foci in the conization specimen were not significant. The power of this study to detect a 25% difference in the risk of residual invasion was 73% for depth of invasion and 75% for number of invasive foci. CONCLUSION: Women with microinvasive squamous carcinoma in a conization specimen in which both the internal conization margin and post-conization ECC are negative have a low risk of residual invasion and are candidates for follow-up or simple hysterectomy. If either the internal margin or the post-conization ECC contains dysplasia or carcinoma, the risk of residual invasion is high and warrants repeat conization before definitive treatment planning. PMID- 9351761 TI - Basal cell carcinoma of the vulva: clinical features and treatment results in 28 patients. AB - OBJECTIVE: To review our experience and that in the recent literature regarding basal cell carcinoma of the vulva to see whether current management guidelines are appropriate. METHODS: Twenty-eight women with basal cell carcinoma of the vulva were seen over 25 years at the BC Cancer Agency. The clinical-pathologic features were tabulated and the outcome was analyzed. RESULTS: The mean age was 74 years, and almost two-thirds were over the age of 70 at diagnosis. Patients typically presented with an irritation or soreness, with a symptom duration ranging from a few months to several years. Most lesions were confined to the anterior half of the vulva, and 23 of the 28 patients had T1 lesions. Wide local excision was the treatment method used most commonly. Only one patient was known to have died from disease metastasis. Ten women had other basal cell carcinomas, either before or after the diagnosis of their vulvar lesions, and in ten patients 11 other malignancies were diagnosed. CONCLUSION: Basal cell carcinoma of the vulva is an extremely uncommon tumor that rarely metastasizes or spreads. Primary treatment should consist of wide local excision and continued follow-up. PMID- 9351762 TI - Written patient information about triple-marker screening: a randomized, controlled trial. AB - OBJECTIVE: To investigate to what extent a newly revised educational pamphlet on triple-marker screening improves patient knowledge and to identify subgroups of women who may not benefit from these materials. METHODS: Women in six geographically and demographically diverse Ontario sites were allocated randomly to receive the pamphlet on triple-marker screening or a similar-appearing educational pamphlet on daily activities during pregnancy. The primary outcome measure was the Maternal Serum Screening Knowledge Questionnaire, a previously validated 14-item scale. RESULTS: Baseline demographic, obstetric, and medical factors were comparable in the intervention and control groups, as were measures of previous exposure to triple-marker screening. Knowledge scores were significantly higher among the 133 women receiving the intervention pamphlet than among 64 women who received the control pamphlet (0.89 versus 0.52 on a scale from -2 to +2, P < .001). Subgroups not benefiting from the pamphlet on triple marker screening were women age 25 and younger and those not speaking English at home. Those who had completed university or postgraduate education had high levels of knowledge with and without the pamphlet. CONCLUSION: Written patient information can contribute in an important way to patient knowledge about triple marker screening. Providers of antenatal care should be made aware of the value of written patient information as well as the limitations for some subgroups of women. These subgroups are likely to require additional educational materials and resources. It would be appropriate to make these materials available to the general public and pregnant women in their physicians' offices. PMID- 9351763 TI - A randomized placebo-controlled evaluation of terbutaline for external cephalic version. AB - OBJECTIVE: To evaluate the efficacy of subcutaneous terbutaline therapy on the success rate of external cephalic version in term gestation. METHODS: Women with singleton noncephalic gestations were assigned randomly to receive either terbutaline (0.25 mg) or placebo. Physicians were blinded to the assignment. Fifteen to 30 minutes after the study drug was administered, external cephalic version was attempted. It was discontinued after three attempts, for patient discomfort, for fetal heart rate decelerations, or when successful. Patients were discharged home after the procedure and allowed to enter spontaneous labor. Primary outcomes evaluated included initial success of version, presentation in labor, and route of delivery. RESULTS: One hundred three women were enrolled in the study between January 1994 and June 1995, of whom 52 were assigned to terbutaline and 51 to placebo. External cephalic version was successful in 27 of 52 (52%) women receiving terbutaline compared with 14 of 51 (27%) of those receiving placebo (P = .019). This comparison yielded a relative risk (RR) of 1.9 (95% confidence interval [CI] 1.3, 6.5). Four of the 27 (15%) successful versions in the terbutaline group and three of the 14 (21%) successful versions in the placebo group spontaneously reverted to breech presentation. Ultimately, in labor there were 24 (46%) cephalic presentations in the terbutaline group and 13 (25%) in the placebo group (P = .048, RR 1.84, 95% CI 1.1, 5.8). Cesarean delivery rates were 11 of 41 (27%) for women with successful versions and 58 of 62 (94%) among those with failed versions (P < .001). CONCLUSION: Terbutaline (0.25 mg) administered subcutaneously before an attempted version in women at term with noncephalic presentations significantly increased the initial success rate of version and the rate of cephalic presentations in labor while decreasing the rate of cesarean delivery. PMID- 9351764 TI - Maternal insulin sensitivity and cord blood peptides: relationships to neonatal size at birth. AB - OBJECTIVE: To examine the relationship of multiple maternal and cord blood correlates of newborn size to determine the relative strength of the insulin-like growth factor-I association. METHODS: Thirty-seven venous cord blood specimens were obtained at the time of delivery. Ponderal index and birth weight percentile were calculated at birth. Neonatal length estimates were performed with a measuring board. All mothers were nonsmokers and had normal glucose tolerance. There was a wide range of maternal prepregnancy body mass indexes (BMI) (19.6 43.4). Neonates had a wide range of ponderal indexes (2.12-2.75) and birth weight percentiles (7-99th percentile). Univariate correlation coefficients were calculated to determine simple relationships. Stepwise linear regression analyses were performed to determine the relative contribution of potential explanatory variables to both ponderal index and birth weight percentile. Potentially explanatory independent variables included maternal prepregnancy BMI, weight gain in pregnancy, and maternal insulin sensitivity at 32 weeks' gestation. Maternal insulin sensitivity was estimated using the minimal model technique. Neonatal variables included sex, cord blood albumin, insulin, insulin-like growth factor I, insulin-like growth factor-binding protein-1, and insulin-like growth factor binding protein-3. RESULTS: Significant positive univariate correlations were identified between cord blood insulin-like growth factor-I and insulin-like growth factor-binding protein-3 with neonatal ponderal index and birth weight percentile. Maternal insulin sensitivity demonstrated a negative correlation with birth weight percentile (r = -.35, P < .05). Cord blood insulin correlated positively with birth weight percentile (r = .32, P < .05). There were no significant associations of cord blood insulin-like growth factor-binding protein 1 or albumin with either index of newborn size. Stepwise logistic regression analysis demonstrated an independent association of insulin-like growth factor-I with ponderal index (r2 = .41, P < .001). Both insulin-like growth factor-I and male sex were associated independently with birth weight percentile (r2 = .38, P < .001). No additional independent variables contributed to the prediction of ponderal index or birth weight percentile. CONCLUSION: These data support a unique relationship between cord blood insulin-like growth factor-I and newborn size under normal growth conditions. This is manifest by the strength and independence of the association between insulin-like growth factor-I and neonatal birth weight percentile ponderal index. PMID- 9351765 TI - The value of the cervical score in predicting successful outcome of labor induction. AB - OBJECTIVE: To compare cervical dilation and the Bishop score as correlates of successful labor induction and vaginal delivery and to determine whether the prognosis of post-ripening cervical characteristics varies with the method of ripening used. METHODS: Four hundred forty-three women with Bishop scores less than 9 who required induction of labor were assigned randomly to cervical ripening with prostaglandin E2 gel or hygroscopic dilation. The Bishop score and its component characteristics were evaluated as univariate correlates of successful induction of labor and vaginal delivery and then were assessed using logistic regression to adjust for other maternal and fetal factors. The differences in the association between method of ripening and successful labor induction were evaluated relative to pre-ripening and post-ripening cervical examination characteristics. RESULTS: Cervical dilation was a better correlate of successful labor induction and vaginal delivery than was the Bishop score, even after exclusion of patients with initial Bishop scores greater than 6 and dilation greater than 3.0. Both ripening methods yielded similar success in labor induction and vaginal delivery, but when categorized by post-ripening cervical examinations, patients undergoing hygroscopic ripening had lower rates of successful labor induction and vaginal delivery. CONCLUSION: Cervical dilation is a better predictor of successful labor induction and vaginal delivery than either the Bishop score or any other Bishop score component characteristic. The likelihood of successful labor induction and vaginal delivery based on post ripening cervical characteristics varies by the ripening method used. PMID- 9351766 TI - Illicit use of clonidine in opiate-abusing pregnant women. AB - OBJECTIVE: To examine prevalence rates and psychosocial correlates of clonidine use in a sample of opiate-dependent pregnant women. METHODS: Clonidine use was assessed in 90 treatment-seeking, pregnant, opiate-abusing women using both self report and urinalysis toxicology. Clonidine-positive and -negative subjects were compared for selected demographic, substance use, and psychosocial measures. RESULTS: One-third of the sample was clonidine-positive. Urinalysis identified 26 clonidine-positive subjects, whereas self-report detected only six cases. Logistic regression identified four predictors of clonidine use at treatment admission: recent clinical anxiety, greater severity of family or social problems, recent cocaine use, and recent drug treatment. CONCLUSION: Clonidine use is prevalent in treatment-seeking opiate abusers, particularly those with concurrent cocaine use. The abuse potential of the drug warrants further study in this high-risk population. PMID- 9351767 TI - Effect of long-term cocaine administration to pregnant ewes on fetal hemodynamics, oxygenation, and growth. AB - OBJECTIVE: To assess uterine and fetal blood flows by Doppler velocimetry and fetal growth and oxygenation in pregnant ewes treated daily with cocaine and to determine whether cocaine impairs fetal cardiac and cerebral reactivity. METHODS: The study groups received 70 mg (n = 7) or 140 mg (n = 7) of cocaine and the control group (n = 7) received placebo injected intramuscularly daily on days 60 134. Hemodynamic data were measured at rest and during two acute hypoxic tests at cesarean delivery performed on day 134. RESULTS: The fetal heart rate (FHR) and umbilical and uterine resistance indices (RIs) were higher in the cocaine groups than in the control group (FHR: 187 +/- 8 and 166 +/- 8 beats per minute at 83 and 123 days, respectively, in controls and 9-11% higher in cocaine groups; umbilical RI: 0.79 +/- 0.06, 0.60 +/- 0.04, and 0.52 +/- 0.06, at 83, 105, and 123 days, respectively, in controls and 11-17% higher in the cocaine groups [P < .01]; and uterine RI: 0.40 +/- 0.05, 0.40 +/- 0.04, and 0.37 +/- 0.04, at 83, 105, and 123 days, respectively, in controls and 13-35% higher in cocaine groups [P < .05]). At delivery on day 134, the following characteristics were found to be different in the cocaine groups: fetal weight (4.03 +/- 0.2 kg in controls and 15-21% lower in the cocaine groups [P < .02]), partial pressure of oxygen (26.5 +/- 1.4 mmHg in controls and 15-16% lower in cocaine groups [P < .05]), umbilical RI (0.40 +/- 0.03 in controls and 11-17% higher in cocaine groups [P < .01]), cerebral RI (0.61 +/- 0.03 in controls and 9-15% lower in cocaine groups [P < .01]), and cerebral-umbilical ratio (1.52 +/- 0.04 in controls and 22-23% lower in cocaine groups [P < .001]). During the hypoxic tests, the cerebral RI (P < .05) and the cerebral-umbilical ratio (P < .05) decreased significantly less in the two cocaine groups. The FHR response was reduced significantly in the two cocaine groups (P < .05). CONCLUSION: Long-term exposure to cocaine induces uterine and fetal blood flow disorders, fetal growth restriction, and hypoxia. It reduces the capability of the cerebral vessels to vasodilate and the heart rate to increase during acute hypoxia. PMID- 9351768 TI - Outcome of infants born at 24-26 weeks' gestation: I. Survival and cost. AB - OBJECTIVE: To determine neonatal survival, short-term morbidities, and cost per survivor in pregnancies delivered at 24-26 weeks' gestation in a center in which antenatal steroids and exogenous surfactant are standard care. METHODS: A retrospective cohort study compared survival, short-term outcome, and initial hospital charges for pregnancies delivered at 24-26 weeks during 1990-1994. We calculated hospital costs for each year by using the corresponding institutional cost-charge ratio. RESULTS: There were 138 infants after excluding those with severe anomalies. Survival was 43%, 74%, and 83% at 24, 25, and 26 weeks, respectively (P = .006). The majority of women received antenatal steroids, and the majority of surviving neonates received exogenous surfactant. Severe retinopathy of prematurity and chronic lung disease decreased significantly from 24 to 26 weeks (P < or = .026). The likelihood of having a surviving infant without chronic lung disease or severe retinopathy of prematurity was 35% at 24 weeks and 78% at 26 weeks. Hospital costs for the 29 nonsurvivors were $1.46 million and for the 94 surviving infants were $16.9 million. The cost per day was similar at each gestational age, whereas the cost to produce a survivor was $294,749, $181,062, and $166,215 at 24, 25, and 26 weeks, respectively. CONCLUSION: Survival at 24 weeks was only 43% despite treatment with antenatal steroids and exogenous surfactant. The cost per survivor for infants born at 24 weeks was higher than the cost for those born after 1 more week in utero. Outcome improved markedly between 24 and 26 weeks, and small differences in gestational age lead to large economic differences. All efforts should be attempted to prolong pregnancy, and if prolongation is unsuccessful, treatment options including nonintervention should be available to parents of 24-week gestations. PMID- 9351769 TI - Outcome of infants born at 24-26 weeks' gestation: II. Neurodevelopmental outcome. AB - OBJECTIVE: To assess the neurodevelopmental outcome of infants born at 24-26 weeks' gestation. METHODS: One hundred thirty-eight nonanomalous infants were born at our hospital after pregnancies of 24-26 weeks' gestation between 1990 and 1994. Ninety-four infants survived to discharge and 86 were followed in a nursery follow-up program for outcome. Associations between gestational age and neurodevelopmental outcome and risk factors and outcome were analyzed. Mean age at follow-up was 32 months. RESULTS: The frequency of cerebral palsy did not differ significantly in the three groups (11, 20, and 11% at 24, 25, and 26 weeks, respectively). The incidence of normal cognitive outcome was associated significantly with gestational age at birth (28, 47, and 71% normal at 24, 25, and 26 weeks, respectively). Poor neurologic outcome was associated with the medical risk factor of intracranial hemorrhage grade 3 or 4 or periventricular leukomalacia. Poor cognitive outcome was correlated with both medical and social risk factors; however, there was an association between poor cognitive outcome and lower gestational age (P < .05), regardless of the relationships of any other risk factors to cognitive outcome. CONCLUSION: Although the incidence of cerebral palsy was low in these three groups, the high percentage of infants born at 24 and 25 weeks' gestation with cognitive deficits is concerning. PMID- 9351770 TI - Umbilical cord blood interleukin-6 levels and neonatal morbidity. AB - OBJECTIVE: To study umbilical cord interleukin-6 levels and the occurrence of neonatal sepsis, congenital pneumonia, necrotizing enterocolitis, and grade II-IV intraventricular hemorrhage. METHODS: Umbilical cord blood was collected from 133 preterm newborns. The study population was divided according to the presence or absence of neonatal complications. Interleukin-6 levels and clinical characteristics were compared by univariate and multivariate analyses. RESULTS: Sixteen neonates had adverse outcomes, and 117 were unaffected. The median interleukin-6 level was significantly higher in affected than in unaffected infants (145 pg/mL versus 0 pg/mL, P = .002). Elevated interleukin-6 levels were associated independently with neonatal morbidity in multiple logistic regression modeling that included gestational age, birth weight, and antenatal steroid exposure. CONCLUSION: Umbilical cord blood interleukin-6 levels are elevated in neonates who subsequently develop sepsis, congenital pneumonia, necrotizing enterocolitis, or grade II-IV intraventricular hemorrhage. PMID- 9351772 TI - Cost-effectiveness of fetal lung maturity testing in preterm labor. AB - OBJECTIVE: To determine the marginal cost-effectiveness of two strategies for preventing respiratory distress syndrome (RDS) resulting from preterm birth: 1) tocolysis with beta-mimetic agonists and treatment with corticosteroids (TREATALL), and 2) amniocentesis and testing for fetal lung maturity, with treatment based on test results (TESTALL), compared with no treatment. METHODS: We used a Markov decision analytic model to estimate the outcomes of each strategy, from a hospital-based perspective. Probability variables were obtained from the literature, whereas cost variables came from the Beth Israel-Deaconess Medical Center. Sensitivity analysis was performed on all variables. RESULTS: The most cost-effective strategy varied with the probability of RDS. TREATALL was the most cost-effective strategy above a probability of 17% (before 34 weeks' gestation), TESTALL was most cost-effective from 17% to 2% (34-36 weeks), and it was most cost-effective to use no treatment at probabilities less than 2% (after 36 weeks). TREATALL and TESTALL were both cost-saving compared with no treatment at probabilities of RDS above 2%. TREATALL was more highly favored as the costs of RDS and preterm birth increased, whereas TESTALL was more favored as the specificity of the test and the cost of maternal hospitalization increased. CONCLUSION: Although testing for fetal lung maturity is useful in many clinical situations, the cost-effectiveness of such testing in the setting of idiopathic preterm labor from a tertiary medical center perspective depends primarily on the probability and costs of RDS and the costs of non-RDS-related morbidity. At our institution, such testing is cost-effective between 34 and 36 weeks' gestation. PMID- 9351771 TI - Neonatal effects and serum cortisol levels after multiple courses of maternal corticosteroids. AB - OBJECTIVE: To determine the effects of multiple courses of maternal betamethasone for fetal lung maturation on neonatal serum cortisol levels and clinical Cushing syndrome. METHODS: Seventy-nine mother-infant pairs delivered between 24 and 36 weeks' gestation were enrolled in the study. They were grouped according to the number of courses of betamethasone received between 24 and 34 weeks' gestation for fetal lung maturation: those receiving no courses, one course, and two or more courses. Physical examinations were performed and serum glucose, electrolyte, and cortisol levels were measured on days 1 and 3 of life. RESULTS: For those receiving multiple courses of betamethasone (n = 43), the mean (+/- standard error of the mean [SEM]) number of courses was 5.3 +/- 0.4, with a mean (+/-SEM) total dose of 125.0 +/- 10.7 mg. No neonates had findings suggestive of Cushing syndrome. Day 1 cortisol levels (pooled mean +/- SEM) were 12.6 +/- 2.4, 5.3 +/- 3.2, and 4.4 +/- 1.8 microg/dL in those receiving no courses, one course, and two or more courses, respectively (P = .03; no courses versus two or more courses, P = .03), but the differences were not significant when corrected for multiple variables. Differences among day 3 cortisol levels (pooled mean +/- SEM) were not significant: 8.3 +/- 1.6, 5.8 +/- 1.4, and 5.8 +/- 0.9 microg/dL in those receiving no courses, one course, and two or more courses, respectively. None of the neonates in the group receiving no courses of betamethasone had day 1 cortisol levels lower than normal, whereas 22% and 11% of the neonates receiving one and two or more courses, respectively, had day 1 levels lower than normal. On day 3, 15% of those receiving one course and 10% of those receiving two or more courses had serum cortisol levels lower than normal, whereas none of those who received no courses had a low cortisol level. Multivariate regression analysis could show no association between the number of courses or total dose of betamethasone and the day 1 or day 3 cortisol values. The day 1 cortisol level (log10) was most associated with the severity of respiratory distress syndrome (RDS) and day 3 cortisol level (log10) with race and severity of RDS. Only in neonates with absent or mild RDS did number of courses correlate with day 3 cortisol levels (log10), but this was a positive correlation. CONCLUSION: Serum cortisol levels either were independent of the number of courses or total dose of corticosteroids given or, in a subpopulation, were associated with increasing levels with increasing doses, suggesting that there is no suppressive effect with repeated dosing. PMID- 9351773 TI - Placental pathology of absent and reversed end-diastolic flow in growth restricted fetuses. AB - OBJECTIVE: To identify placental histopathology associated with absent and reversed end-diastolic flow demonstrated by umbilical artery (UA) Doppler velocimetry in fetal growth restriction (FGR). METHODS: Between January 1989 and June 1995, 64 consecutive, nonanomalous singletons at less than the tenth percentile for birth weight were admitted to the neonatal intensive care unit, with UA Doppler velocimetry obtained within 3 days of delivery; 54 of the 64 (84%) had placental histopathology. Umbilical artery Doppler wave forms were classified as having end-diastolic flow (n = 26), and either absent (n = 20) or reversed end-diastolic flow (n = 8). Blinded review of placental histology scored lesions in categories of intraplacental vaso-occlusion, uteroplacental vascular pathology, chronic inflammation, and coagulation. RESULTS: Using cases of FGR with end-diastolic flow present as the control population, we found that absent end-diastolic flow cases had significantly more fetal stem vessels with medial hyperplasia and luminal obliteration, and cases of reversed end-diastolic flow had significantly more poorly vascularized terminal villi, villous stromal hemorrhage, "hemorrhagic endovasculitis," and abnormally thin-walled fetal stem vessels (each P < .005). CONCLUSION: In FGR, UA Doppler velocity wave forms do not demonstrate a continuum of placental lesions in which reversed end-diastolic flow reflects more severe placental histopathology than absent end-diastolic flow and end-diastolic flow present. As expected, absent end-diastolic flow cases had more occlusive lesions of the intraplacental vasculature. In reversed end diastolic flow, lesions suggesting vascular remodeling and/or damage by pathologic conditions of intraplacental flow predominated. PMID- 9351774 TI - Digital communication with fetal monitors. AB - BACKGROUND: Fetal heart rate (FHR) values in the averaged format that are provided by commercial computed cardiotocography analysis systems may be unsuitable for special analysis purposes. METHOD: I developed a communication software program to obtain any measured values of fetal monitors for individual analysis of computed cardiotocography. EXPERIENCE: The software program was used to study the data continuity of beat-to-beat FHR values as an experiment for chaos theory and power spectrum analysis. The results indicated that the signal loss was recognized at a precision of 95%. CONCLUSION: The described method of digital communication with fetal monitors was found to be useful for individual purposes in the field of computed cardiotocography analysis. PMID- 9351775 TI - Cross-sectional imaging anatomy of the anal sphincters. AB - BACKGROUND: To describe the cross-sectional anatomy of the anal sphincter mechanism relevant to magnetic resonance imaging (MRI) and ultrasound cross sectional images. METHOD: Axial, sagittal, and coronal 5-mm sections of female pelves were reviewed from six cadaver specimens (ages 24-72 years). Fetal anatomy was studied in plastinated histologic sections from 19 and 26 weeks' gestation. Images of the anal sphincter were obtained by MRI in six and by ultrasound using an exoanal technique in 12 nulliparous volunteers. EXPERIENCE: The internal anal sphincter is clearly visible in anatomic sections central to the external sphincter and is visible in MRI and ultrasound images. The external anal sphincter can be subdivided into a subcutaneous and a deep portion. On anatomic sections and on MRI, the subcutaneous part shows as two parallel muscle strips in the axial plane; the deep portion presents with a characteristic teardrop form in the section perpendicular to the axis of the anal canal. The puborectalis muscle and the external anal sphincter form a "double bump" in the sagittal section. The longitudinal muscle can be identified by its fiber orientation in anatomic sections but is not clearly visible in imaging studies. CONCLUSION: This information should make it possible to identify accurately anal sphincter anatomy in two-dimensional sectional images of the anal sphincter. PMID- 9351776 TI - Currycombs for the vaginal paravaginal defect repair. AB - BACKGROUND: The paravaginal defect, present in more than three-quarters of patients with cystoceles, can be repaired by both the abdominal and vaginal approaches. The technical challenges of the vaginal paravaginal repair have militated against its widespread adoption by gynecologic surgeons. INSTRUMENT: Currycombs can be used to facilitate suture management during vaginal paravaginal repair. EXPERIENCE: The vaginal paravaginal repair using currycombs was performed as part of pelvic repair surgery on 27 patients. Perioperative complications were minimal. A cystocele cure rate of 80% was achieved after a mean follow-up of 8 months. CONCLUSION: The use of currycombs during performance of the vaginal paravaginal repair facilitates suture management. The addition of this technique should help gynecologic surgeons to perform this somewhat daunting surgical procedure. PMID- 9351777 TI - Antenatal screening for factor V Leiden mutation: a critical appraisal. AB - Thromboembolic disease is a leading cause of maternal mortality in the United States. Recently, inherited resistance to activated protein C has been recognized as a major risk factor for thrombosis and has been demonstrated in 20-60% of patients with clinically evident thrombosis. The factor V Leiden mutation, which is readily detectable by molecular DNA techniques, is responsible for 90-95% of cases of activated protein C resistance. Because 5% of whites and 1% of blacks in the United States are heterozygous for the Leiden mutation, at least one group has suggested that screening of asymptomatic gravidas for the mutation should be considered. Therefore, we conducted a combined MEDLINE and bibliographic literature search for relevant data and evaluated screening for the factor V Leiden mutation in the context of well-elucidated desirable characteristics for a successful screening program. Based on this evaluation, we conclude that routine antenatal screening for the factor V Leiden mutation cannot be recommended at the present time. PMID- 9351778 TI - Antenatal corticosteroids in pregnancies complicated by preterm premature rupture of membranes. AB - In 1994, the National Institutes of Health Consensus Development Conference on Antenatal Steroids recommended corticosteroids between 24 and 30-32 weeks' gestation in pregnancies complicated by preterm premature rupture of membranes (PROM). Since the Consensus Conference, the use of antenatal corticosteroids has increased to approximately 60% of potential treatment candidates. Some of the remaining 40% of pregnant candidates may go untreated because of concern that corticosteroids could increase the risk of neonatal infection. Using decision analysis techniques, we compared the potential benefit of antenatal corticosteroids in reducing the incidence of severe intraventricular hemorrhage with the potential risk of increasing the rate of neonatal sepsis. Our analysis indicates that the benefit of a small decrease in severe intraventricular hemorrhage outweighs the potential harm of a large increase in the rate of neonatal sepsis. Therefore, we support the Consensus Conference panel's recommendation that antenatal corticosteroids be used in pregnancies complicated by preterm PROM. PMID- 9351779 TI - Analysis of the effectiveness of an endoscopy education program in improving residents' laparoscopic skills. AB - OBJECTIVE: To evaluate the effectiveness of a gynecologic endoscopy education program in enhancing residents' proficiency in laparoscopic surgery. METHODS: The program was designed to provide residents with the knowledge and skills necessary for laparoscopic surgery, before entering the operating room, in a cost-effective manner that honored the principles of adult education. The 7-week program included didactic sessions to provide conceptual learning but focused on practical skills enhancement through practice in both pelvic trainer and animal laboratory settings. The program design included dominant, nondominant, and two handed skills as well as models for laparoscopic dissection, hemostasis, and suturing. The evaluation of the program is based on timing of laparoscopic skills as well as resident and faculty evaluation of laparoscopic proficiency at the beginning and end of the program. RESULTS: Prior to the program, 48% of residents and 75% of faculty were satisfied with laparoscopic training. All residents improved operating times in pelvic trainer skills after the program, with first year residents improving by 68%, third-year residents by 58%, and fourth-year residents by 72%. The residents self-assessment of competence in 14 laparoscopic skills revealed an increase in all skills following the program. The faculty assessment showed an upward trend in skills competence. At the conclusion of the program, 100% of residents and 92% of faculty were satisfied with the laparoscopic training. CONCLUSION: A structured program emphasizing skills enhancement is an effective approach to improve residents' performance in laparoscopic surgery. PMID- 9351780 TI - The safety of early postpartum discharge: a review and critique. AB - OBJECTIVE: To determine the effect of early postpartum discharge (less than 48 hours after vaginal birth or 96 hours after cesarean delivery) on maternal and neonatal complications, maternal concerns, patient satisfaction, and cost savings. DATA SOURCES: We performed a MEDLINE search of English-language journals for pertinent articles published from 1966 through January 1997. We also reviewed reference lists in all the articles retrieved in the search as well as those of major obstetric texts. METHODS OF STUDY SELECTION: We included all studies describing early postpartum discharge. TABULATION, INTEGRATION, AND RESULTS: Studies included five randomized controlled trials, ten cohort studies, one case control study, and 12 case-series reports. We classified the data using the rating system of the U.S. Preventive Services Task Force. We calculated relative risks and 95% confidence intervals for maternal and neonatal readmission and outpatient treatment after early postpartum discharge. Most studies did not show an increase in maternal or neonatal morbidity after early discharge. The five randomized controlled studies did not meet criteria for properly designed trials. Most evidence consists of cohort studies and case-series (class II-2 and III evidence) of highly selected patients with extensive supplemental antepartum and postpartum care and education. CONCLUSION: The current data do not support or condemn widespread use of early postpartum discharge in the general population (class C recommendation). Early postpartum discharge appears safe for carefully selected, consenting patients. Whether these data can be extrapolated to the general population of pregnant women remains unknown. PMID- 9351781 TI - A study of ruptured tubal ectopic pregnancy. PMID- 9351801 TI - Structure of mouse 7S NGF: a complex of nerve growth factor with four binding proteins. AB - BACKGROUND: Nerve growth factor (NGF) is a neurotrophic factor that promotes the differentiation and survival of certain populations of neurons in the central and peripheral nervous systems. 7S NGF is an alpha 2 beta 2 gamma 2 complex in which the beta-NGF dimer (the active neurotrophin) is associated with two alpha-NGF and two gamma-NGF subunits, which belong to the glandular kallikrein family of serine proteinases. The gamma-NGF subunit is an active serine proteinase capable of processing the precursor form of beta-NGF, whereas alpha-NGF is an inactive serine proteinase. The structure of 7S NGF could be used as a starting point to design inhibitors that prevent NGF binding to its receptors, as a potential treatment of neurodegenerative diseases. RESULTS: The crystal structure of 7S NGF shows that the two gamma-NGF subunits make extensive interactions with each other around the twofold axis of the complex and have the C-terminal residues of the beta-NGF subunits bound within their active sites. The 'activation domain' of each of the alpha-NGF subunits is in an inactive (zymogen-like) conformation and makes extensive interactions with the beta-NGF dimer. The two zinc ions that stabilize the complex are located at the relatively small interfaces between the alpha-NGF and gamma-NGF subunits. CONCLUSIONS: The structure of 7S NGF shows how the twofold axis of the central beta-NGF dimer organizes the symmetry of this multisubunit growth factor complex. The extensive surface of beta-NGF buried within the 7S complex explains the lack of neurotrophic activity observed for 7S NGF. The regions of the beta-NGF dimer that contact the alpha-NGF subunits overlap with those known to engage NGF receptors. Two disulphide-linked loops on alpha-NGF make multiple interactions with beta-NGF and suggest that it might be possible to design peptides that inhibit the binding of beta-NGF to its receptors. PMID- 9351802 TI - La cage aux fold: asymmetry in the crystal structure of GroEL-GroES-(ADP)7. AB - The structure of the molecular chaperone GroEL from Escherichia coli in complex with GroES and seven ADP molecules has recently been reported to 3 A resolution. The structure illustrates how the cavity of GroEL is converted from a hydrophobic environment, suitable for binding unfolded polypeptides, to a much larger hydrophilic environment suitable for refolding proteins. PMID- 9351803 TI - Structures of class pi glutathione S-transferase from human placenta in complex with substrate, transition-state analogue and inhibitor. AB - BACKGROUND: Glutathione S-transferases (GSTs) are detoxification enzymes, found in all aerobic organisms, which catalyse the conjugation of glutathione with a wide range of hydrophobic electrophilic substrates, thereby protecting the cell from serious damage caused by electrophilic compounds. GSTs are classified into five distinct classes (alpha, mu, pi, sigma and theta) by their substrate specificity and primary structure. Human GSTs are of interest because tumour cells show increased levels of expression of single classes of GSTs, which leads to drug resistance. Structural differences between classes of GST can therefore be utilised to develop new anti-cancer drugs. Many mutational and structural studies have been carried out on the mu and alpha classes of GST to elucidate the reaction mechanism, whereas knowledge about the pi class is still limited. RESULTS: We have solved the structures of the pi class GST hP1-1 in complex with its substrate, glutathione, a transition-state complex, the Meisenheimer complex, and an inhibitor, S-(rho-bromobenzyl)-glutathione, and refined them to resolutions of 1.8 A, 2.0 A and 1.9 A, respectively. All ligand molecules are well-defined in the electron density. In all three structures, an additionally bound N-morpholino-ethansulfonic acid molecule from the buffer solution was found. CONCLUSIONS: In the structure of the GST-glutathione complex, two conserved water molecules are observed, one of which hydrogen bonds directly to the sulphur atom of glutathione and the other forms hydrogen bonds with residues around the glutathione-binding site. These water molecules are absent from the structure of the Meisenheimer complex bound to GST, implicating that deprotonation of the cysteine occurs during formation of the ternary complex which involves expulsion of the inner bound water molecule. The comparison of our structures with known mu class GST structures show differences in the location of the electrophile-binding site (H-site), explaining the different substrate specificities of the two classes. Fluorescence measurements are in agreement with the position of the N-morpholino-ethansulfonic acid, close to Trp28, identifying a possible ligandin-substrate binding site. PMID- 9351804 TI - The role of oil in macromolecular crystallization. AB - The different facets of the utilization of oil demonstrate that an individual oil and/or combinations of different oils can influence the outcome of crystallization experiments. The oil can play a part in the control of nucleation, affect the rate of equilibration and consequently determine the size of the forming crystals. Whether used for microbatch, vapour diffusion or for control of nucleation, the presence of oil is a parameter that can contribute to the accuracy, cleanliness and to the increase in the reproducibility of the experiments. Furthermore, the oil has a role in the protection of the trials during the course of their duration and in maintaining the stability of the resulting crystals. PMID- 9351805 TI - Insight into the stabilization of A-DNA by specific ion association: spontaneous B-DNA to A-DNA transitions observed in molecular dynamics simulations of d[ACCCGCGGGT]2 in the presence of hexaamminecobalt(III). AB - BACKGROUND: Duplex DNA is more than a simple information carrier. The sequence dependent structure and its inherent deformability, in concert with the subtle modulating effects of the environment, play a crucial role in the regulation and packaging of DNA. Recent advances in force field and simulation methodologies allow molecular dynamics simulations to now represent the specific effects of the environment. An understanding of the environmental dependence of DNA structure gives insight into how histones are able to package DNA, how various proteins are able to bind and modulate nucleic acid structure and will ultimately aid the design of molecules to package DNA for more effective gene therapy. RESULTS: Molecular dynamics simulations of d[ACCCGCGGGT]2 in solution in the presence of hexaamminecobalt(III) [Co(NH3)6(3+)] show stabilization of A-DNA and spontaneous B-DNA to A-DNA transitions, which is consistent with experimental results from NMR and Raman spectroscopic and X-ray crystallographic studies. In the absence of Co(NH3)6(3+), A-DNA to B-DNA transitions are observed instead. In addition to their interaction with the guanines in the major groove, Co(NH3)6(3+) ions bridge opposing strands in the bend across the major groove, probably stabilizing A-DNA. CONCLUSIONS: The simulation methods and force fields have advanced to a sufficient level that some representation of the environment can be seen in nanosecond length molecular dynamics simulations. These simulations suggest that, in addition to the general explanation of A-DNA stabilization by dehydration, hydration and ion association in the major groove stabilize A-DNA. PMID- 9351806 TI - The SH2 domain from the tyrosine kinase Fyn in complex with a phosphotyrosyl peptide reveals insights into domain stability and binding specificity. AB - BACKGROUND: SH2 domains are found in a variety of signal transduction proteins; they bind phosphotyrosine-containing sequences, allowing them to both recognize target molecules and regulate intramolecular kinase activity. Fyn is a member of the Src family of tyrosine kinases that are involved in signal transduction by association with a number of membrane receptors. The kinase activity of these signalling proteins is modulated by switching the binding mode of their SH2 and SH3 domains from intramolecular to intermolecular. The molecular basis of the signalling roles observed for different Src family members is still not well understood; although structures have been determined for the SH2 domains of other Src family molecules, this is the first structure of the Fyn SH2 domain. RESULTS: The structure of the Fyn SH2 domain in complex with a phosphotyrosyl peptide (EPQpYEEIPIYL) was determined by high resolution NMR spectroscopy. The overall structure of the complex is analogous to that of other SH2-peptide complexes. Noteworthy aspects of the structure are: the BG loop, which contacts the bound peptide, contains a type-I' turn; a capping-box-like interaction is present at the N-terminal end of helix alpha A; cis-trans isomerization of the Val beta G1 Pro beta G2 peptide bond causes conformational heterogeneity of residues near the N and C termini of the domain. CONCLUSIONS: Comparison of the Fyn SH2 domain structure with other structures of SH2 domains highlights several interesting features. Conservation of helix capping interactions among various SH2 domains is suggestive of a role in protein stabilisation. The presence of a type-I' turn in the BG loop, which is dependent on the presence of a glycine residue at position BG3, is indicative of a binding pocket, characteristic of the Src family, SykC and Abl, rather than a binding groove found in PLC-gamma 1C, p85 alpha N and Shc, for example. PMID- 9351807 TI - The crystal structure of vascular endothelial growth factor (VEGF) refined to 1.93 A resolution: multiple copy flexibility and receptor binding. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is an endothelial cell specific angiogenic and vasculogenic mitogen. VEGF also plays a role in pathogenic vascularization which is associated with a number of clinical disorders, including cancer and rheumatoid arthritis. The development of VEGF antagonists, which prevent the interaction of VEGF with its receptor, may be important for the treatment of such disorders. VEGF is a homodimeric member of the cystine knot growth factor superfamily, showing greatest similarity to platelet-derived growth factor (PDGF). VEGF binds to two different tyrosine kinase receptors, kinase domain receptor (KDR) and Fms-like tyrosine kinase 1 (Flt-1), and a number of VEGF homologs are known with distinct patterns of specificity for these same receptors. The structure of VEGF will help define the location of the receptor-binding site, and shed light on the differences in specificity and cross-reactivity among the VEGF homologs. RESULTS: We have determined the crystal structure of the receptor-binding domain of VEGF at 1.93 A resolution in a triclinic space group containing eight monomers in the asymmetric unit. Superposition of the eight copies of VEGF shows that the beta-sheet core regions of the monomers are very similar, with slightly greater differences in most loop regions. For one loop, the different copies represent different snapshots of a concerted motion. Mutagenesis mapping shows that this loop is part of the receptor-binding site of VEGF. CONCLUSIONS: A comparison of the eight independent copies of VEGF in the asymmetric unit indicates the conformational space sampled by the protein in solution; the root mean square differences observed are similar to those seen in ensembles of the highest precision NMR structures. Mapping the receptor-binding determinants on a multiple sequence alignment of VEGF homologs, suggests the differences in specificity towards KDR and Flt-1 may derive from both sequence variation and changes in the flexibility of binding loops. The structure can also be used to predict possible receptor binding determinants for related cystine knot growth factors, such as PDGF. PMID- 9351808 TI - The coupling of light-induced electron transfer and proton uptake as derived from crystal structures of reaction centres from Rhodopseudomonas viridis modified at the binding site of the secondary quinone, QB. AB - BACKGROUND: In a reaction of central importance to the energetics of photosynthetic bacteria, light-induced electron transfer in the reaction centre (RC) is coupled to the uptake of protons from the cytoplasm at the binding site of the secondary quinone (QB). In the original structure of the RC from Rhodopseudomonas viridis (PDB entry code 1PRC), the QB site was poorly defined because in the standard RC crystals it was only approximately 30% occupied with ubiquinone-9 (UQ9). We report here the structural characterization of the QB site by crystallographic refinement of UQ9-depleted RCs and of complexes of the RC either with ubiquinone-2 (UQ2) or the electron-transfer inhibitor stigmatellin in the QB site. RESULTS: The structure of the RC complex with UQ2, refined at 2.45 A resolution, constitutes the first crystallographically reliably defined binding site for quinones from the bioenergetically important quinone pool of biological, energy-transducing membranes. In the UQ9-depleted QB site of the RC structure, refined at 2.4 A resolution, apparently five (and possibly six) water molecules are bound instead of the ubiquinone head group, and a detergent molecule binds in the region of the isoprenoid tail. All of the protein-cofactor interactions implicated in the binding of the ubiquinone head group are also implicated in the binding of the stigmatellin head group. In the structure of the stigmatellin-RC complex, refined at 2.4 A resolution, additional hydrogen bonds stabilize the binding of stigmatellin over that of ubiquinone. The tentative position of UQ9 in the QB site in the original data set (1PRC) was re-examined using the structure of the UQ9-depleted RC as a reference. A modified QB site model, which exhibits greater similarity to the distal ubiquinone-10 (UQ10) positioning in the structure of the RC from Rhodobacter sphaeroides (PDB entry code 1PCR), is suggested as the dominant binding site for native UQ9. CONCLUSIONS: The structures reported here can provide models of quinone reduction cycle intermediates. The binding pattern observed for the stigmatellin complex, where the ligand donates a hydrogen bond to Ser L223 (where 'L' represents the L subunit of the RC), can be viewed as a model for the stabilization of a monoprotonated reduced intermediate (QBH or QBH-). The presence of Ser L223 in the QB site indicates that the QB site is not optimized for QB binding, but for QB reduction to the quinol. PMID- 9351809 TI - The crystal structure of HIV-1 Nef protein bound to the Fyn kinase SH3 domain suggests a role for this complex in altered T cell receptor signaling. AB - BACKGROUND: Human immunodeficiency virus (HIV) Nef protein accelerates virulent progression of acquired immunodeficiency syndrome (AIDS) by its interaction with specific cellular proteins involved in signal transduction and host cell activation. Nef has been shown to bind specifically to a subset of the Src family of kinases. The structures of free Nef and Nef bound to Src homology region 3 (SH3) domain are important for the elucidation of how the affinity and specificity for the Src kinase family SH3 domains are achieved, and also for the development of potential drugs and vaccines against AIDS. RESULTS: We have determined the crystal structures of the conserved core of HIV-1 Nef protein alone and in complex with the wild-type SH3 domain of the p59fyn protein tyrosine kinase (Fyn), at 3.0 A resolution. Comparison of the bound and unbound Nef structures revealed that a proline-rich motif (Pro-x-x-Pro), which is implicated in SH3 binding, is partially disordered in the absence of the binding partner; this motif only fully adopts a left-handed polyproline type II helix conformation upon complex formation with the Fyn SH3 domain. In addition, the structures show how an arginine residue (Arg77) of Nef interacts with Asp 100 of the so-called RT loop within the Fyn SH3 domain, and triggers a hydrogen-bond rearrangement which allows the loop to adapt to complement the Nef surface. The Arg96 residue of the Fyn SH3 domain is specifically accommodated in the same hydrophobic pocket of Nef as the isoleucine residue of a previously described Fyn SH3 (Arg96-->lle) mutant that binds to Nef with higher affinity than the wild type. CONCLUSIONS: The three dimensional structures support evidence that the Nef-Fyn complex forms in vivo and may have a crucial role in the T cell perturbating action of Nef by altering T cell receptor signaling. The structures of bound and unbound Nef reveal that the multivalency of SH3 binding may be achieved by a ligand induced flexibility in the RT loop. The structures suggest possible targets for the design of inhibitors which specifically block Nef-SH3 interactions. PMID- 9351810 TI - The crystal structure of Escherichia coli purine nucleoside phosphorylase: a comparison with the human enzyme reveals a conserved topology. AB - BACKGROUND: Purine nucleoside phosphorylase (PNP) from Escherichia coli is a hexameric enzyme that catalyzes the reversible phosphorolysis of 6-amino and 6 oxopurine (2'-deoxy)ribonucleosides to the free base and (2'-deoxy)ribose-1 phosphate. In contrast, human and bovine PNPs are trimeric and accept only 6 oxopurine nucleosides as substrates. The difference in the specificities of these two enzymes has been utilized in gene therapy treatments in which certain prodrugs are cleaved by E. coli PNP but not the human enzyme. The trimeric and hexameric PNPs show no similarity in amino acid sequence, even though they catalyze the same basic chemical reaction. Structural comparison of the active sites of mammalian and E. coli PNPs would provide an improved basis for the design of potential prodrugs that are specific for E. coli PNP. RESULTS: The crystal structure of E. coli PNP at 2.0 A resolution shows that the overall subunit topology and active-site location within the subunit are similar to those of the subunits from human PNP and E. coli uridine phosphorylase. Nevertheless, even though the overall geometry of the E. coli PNP active site is similar to human PNP, the active-site residues and subunit interactions are strikingly different. In E. coli PNP, the purine- and ribose-binding sites are generally hydrophobic, although a histidine residue from an adjacent subunit probably forms a hydrogen bond with a hydroxyl group of the sugar. The phosphate-binding site probably consists of two main-chain nitrogen atoms and three arginine residues. In addition, the active site in hexameric PNP is much more accessible than in trimeric PNP. CONCLUSIONS: The structures of human and E. coli PNP define two possible classes of nucleoside phosphorylase, and help to explain the differences in specificity and efficiency between trimeric and hexameric PNPs. This structural data may be useful in designing prodrugs that can be activated by E. coli PNP but not the human enzyme. PMID- 9351811 TI - Twists and turns of the nucleosome: tails without ends. AB - The high-resolution structure of a nucleosome core particle gives us our first detailed look at the primary level of eukaryotic DNA organization. The structure reveals the nature of histone-DNA contacts and provides some surprises regarding the histone tails and their possible involvement in higher levels of chromatin organization. PMID- 9351812 TI - The class II MHC protein HLA-DR1 in complex with an endogenous peptide: implications for the structural basis of the specificity of peptide binding. AB - BACKGROUND: Class II major histocompatibility complex (MHC) proteins are cell surface glycoproteins that bind peptides and present them to T cells as part of the mechanism for detecting and responding to foreign material in the body. The peptide-binding activity exhibits allele-specific preferences for particular sidechains at some positions, although the structural basis of these preferences is not understood in detail. We have determined the 2.45 A crystal structure of the human class II MHC protein HLA-DR1 in complex with the tight binding endogenous peptide A2 (103-117) in order to discover peptide-MHC interactions that are important in determining the binding motif and to investigate conformational constraints on the bound peptide. RESULTS: The bound peptide adopts a polyproline II-like conformation and places several sidechains within pockets in the binding site. Bound water molecules mediate MHC-peptide contacts at several sites. A tryptophan residue from the beta 2 'lower' domain of HLA-DR1 was found to project into a pocket underneath the peptide-binding domain and may be important in modulating interdomain interactions in MHC proteins. CONCLUSIONS: The peptide-binding motif of HLA-DR1 includes an aromatic residue at position +1, an arginine residue at position +2, and a small residue at position +6 (where the numbering refers to the normal MHC class II convention); these preferences can be understood in light of interactions observed in the peptide-MHC complex. Comparison of the structure with that of another MHC-peptide complex shows that completely different peptide sequences bind in essentially the same conformation and are accommodated with only minimal rearrangement of HLA-DR1 residues. Small conformational differences that are observed appear to be important in interactions with other proteins. PMID- 9351813 TI - The multi-talented beta-catenin makes its first appearance. AB - beta-catenin plays a central part in cell adhesion as a structural component of the cadherin complex. In a seemingly disparate role, it is also important in embryo patterning, and now has emerged as a leading actor in carcinogenesis. beta catenin achieves its diverse functions by interacting with many partners. The recent structure of the core domain from beta-catenin suggests how this talented molecule can achieve its many functions. PMID- 9351815 TI - Regulation of stability and function of the epithelial Na+ channel (ENaC) by ubiquitination. AB - The epithelial Na+ channel (ENaC), composed of three subunits (alpha beta gamma), plays a critical role in salt and fluid homeostasis. Abnormalities in channel opening and numbers have been linked to several genetic disorders, including cystic fibrosis, pseudohypoaldosteronism type I and Liddle syndrome. We have recently identified the ubiquitin-protein ligase Nedd4 as an interacting protein of ENaC. Here we show that ENaC is a short-lived protein (t1/2 approximately 1 h) that is ubiquitinated in vivo on the alpha and gamma (but not beta) subunits. Mutation of a cluster of Lys residues (to Arg) at the N-terminus of gamma ENaC leads to both inhibition of ubiquitination and increased channel activity, an effect augmented by N-terminal Lys to Arg mutations in alpha ENaC, but not in beta ENaC. This elevated channel activity is caused by an increase in the number of channels present at the plasma membrane; it represents increases in both cell surface retention or recycling of ENaC and incorporation of new channels at the plasma membrane, as determined by Brefeldin A treatment. In addition, we find that the rapid turnover of the total pool of cellular ENaC is attenuated by inhibitors of both the proteasome and the lysosomal/endosomal degradation systems, and propose that whereas the unassembled subunits are degraded by the proteasome, the assembled alpha beta gamma ENaC complex is targeted for lysosomal degradation. Our results suggest that ENaC function is regulated by ubiquitination, and propose a paradigm for ubiquitination-mediated regulation of ion channels. PMID- 9351817 TI - The large subunit of replication factor C is a substrate for caspase-3 in vitro and is cleaved by a caspase-3-like protease during Fas-mediated apoptosis. AB - Caspase-3 is an ICE-like protease activated during apoptosis induced by different stimuli. Poly(ADP-ribose) polymerase (PARP), the first characterized substrate of caspase-3, shares a region of homology with the large subunit of Replication Factor C (RF-C), a five-subunit complex that is part of the processive eukaryotic DNA polymerase holoenzymes. Caspase-3 cleaves PARP at a DEVD-G motif present in the 140 kDa subunit of RF-C (RFC140) and evolutionarily conserved. We show that cleavage of RFC140 during Fas-mediated apoptosis in Jurkat cells and lymphocytes results in generation of multiple fragments. Cleavage is inhibited by the caspase 3-like protease inhibitor Ac-DEVD-CHO but not the caspase-1/ICE-type protease inhibitor Ac-YVAD-CHO. In addition, recombinant caspase-3 cleaves RFC140 in vitro at least at three different sites in the C-terminal half of the protein. Using amino-terminal microsequencing of radioactive fragments, we identified three sites: DEVD723G, DLVD922S and IETD1117A. We did not detect cleavage of small subunits of RF-C of 36, 37, 38 and 40 kDa by recombinant caspase-3 or by apoptotic Jurkat cell lysates. Cleavage of RFC140 during apoptosis inactivates its function in DNA replication and generates truncated forms that further inhibit DNA replication. These results identify RFC140 as a critical target for caspase-3-like proteases and suggest that caspases could mediate cell cycle arrest. PMID- 9351819 TI - Branched O-linked oligosaccharides ectopically expressed in transgenic mice reduce primary T-cell immune responses. AB - Core 2 beta-1,6-N-acetylglucosaminyltransferase, C2GnT, is a key enzyme in O linked oligosaccharide (O-glycan) biosynthesis and the resultant core 2 branch serves as a backbone for additional glycosylation to form oligosaccharide ligands such as sialyl Le(x). Since the expression of C2GnT is highly regulated during T cell development and increases in pathological conditions such as the Wiskott Aldrich syndrome, we have generated transgenic mice overexpressing C2GnT in the T cell lineage. Surprisingly, T lymphocytes in the transgenic mice develop normally, but they exhibit a reduced immune response when assayed by delayed-type hypersensitivity, proliferation upon stimulation and cytokine production. Moreover, T lymphocytes from the transgenic mice adhere much less efficiently to ICAM-1 and fibronectin than do T lymphocytes from non-transgenic mice. These results indicate that overexpression of the core 2 branched O-glycans in T lymphocytes results in reduced immune responses due to impaired cell-cell interaction. Such an impaired immune response may be one of the causes for immunodeficiency in the Wiskott-Aldrich syndrome. PMID- 9351818 TI - Interferon action and apoptosis are defective in mice devoid of 2',5' oligoadenylate-dependent RNase L. AB - 2',5'-Oligoadenylate-dependent RNase L functions in the interferon-inducible, RNA decay pathway known as the 2-5A system. To determine the physiological roles of the 2-5A system, mice were generated with a targeted disruption of the RNase L gene. The antiviral effect of interferon alpha was impaired in RNase L-/- mice providing the first evidence that the 2-5A system functions as an antiviral pathway in animals. In addition, remarkably enlarged thymuses in the RNase L-/- mice resulted from a suppression of apoptosis. There was a 2-fold decrease in apoptosis in vivo in the thymuses and spleens of RNase L-/- mice. Furthermore, apoptosis was substantially suppressed in RNase L-/- thymocytes and fibroblasts treated with different apoptotic agents. These results suggest that both interferon action and apoptosis can be controlled at the level of RNA stability by RNase L. Another implication is that the 2-5A system is likely to contribute to the antiviral activity of interferon by inducing apoptosis of infected cells. PMID- 9351820 TI - Identification of a species-specific inhibitor of glycosylphosphatidylinositol synthesis. AB - Glycosylphosphatidylinositol (GPI)-anchoring represents a mechanism for attaching proteins to the cell surface that is used among all eukaryotes. A common core structure, EthN-P-Man3-GlcN-PI, is synthesized by sequential transfer of sugars and ethanolamine-P to PI and is highly conserved between organisms. We have screened for natural compounds that inhibit GPI-anchoring in yeast and have identified a terpenoid lactone, YW3548, that specifically blocks the addition of the third mannose to the intermediate structure Man2-GlcN-acyIPI. Consistent with the block in GPI synthesis, YW3548 prevents the incorporation of [3H]myo-inositol into proteins, transport of GPI-anchored proteins to the Golgi and is toxic. The compound inhibits the same step of GPI synthesis in mammalian cells, but has no significant activity in protozoa. These results suggest that despite the conserved core structure, the GPI biosynthetic machinery may be different enough between mammalian and protozoa to represent a target for anti-protozoan chemotherapy. PMID- 9351821 TI - SecY and SecA interact to allow SecA insertion and protein translocation across the Escherichia coli plasma membrane. AB - SecA, the preprotein-driving ATPase in Escherichia coli, was shown previously to insert deeply into the plasma membrane in the presence of ATP and a preprotein; this movement of SecA was proposed to be mechanistically coupled with preprotein translocation. We now address the role played by SecY, the central subunit of the membrane-embedded heterotrimeric complex, in the SecA insertion reaction. We identified a secY mutation (secY205), affecting the most carboxyterminal cytoplasmic domain, that did not allow ATP and preprotein-dependent productive SecA insertion, while allowing idling insertion without the preprotein. Thus, the secY205 mutation might affect the SecYEG 'channel' structure in accepting the preprotein-SecA complex or its opening by the complex. We isolated secA mutations that allele-specifically suppressed the secY205 translocation defect in vivo. One mutant protein, SecA36, with an amino acid alteration near the high-affinity ATP binding site, was purified and suppressed the in vitro translocation defect of the inverted membrane vesicles carrying the SecY205 protein. The SecA36 protein could also insert into the mutant membrane vesicles in vitro. These results provide genetic evidence that SecA and SecY specifically interact, and show that SecY plays an essential role in insertion of SecA in response to a preprotein and ATP and suggest that SecA drives protein translocation by inserting into the membrane in vivo. PMID- 9351822 TI - The chaperone-assisted membrane release and folding pathway is sensed by two signal transduction systems. AB - The assembly of interactive protein subunits into extracellular structures, such as pilus fibers in the Enterobacteriaceae, is dependent on the activity of PapD like periplasmic chaperones. The ability of PapD to undergo a beta zippering interaction with the hydrophobic C-terminus of pilus subunits facilitates their folding and release from the cytoplasmic membrane into the periplasm. In the absence of the chaperone, subunits remained tethered to the membrane and were driven off-pathway via non-productive interactions. These off-pathway reactions were detrimental to cell growth; wild-type growth was restored by co-expression of PapD. Subunit misfolding in the absence of PapD was sensed by two parallel pathways: the Cpx two-component signaling system and the sigma E modulatory pathway. PMID- 9351823 TI - MAPK inactivation is required for the G2 to M-phase transition of the first mitotic cell cycle. AB - Down-regulation of MAP kinase (MAPK) is a universal consequence of fertilization in the animal kingdom, although its role is not known. Here we show that MAPK inactivation is essential for embryos, both vertebrate and invertebrate, to enter first mitosis. Suppressing down-regulation of MAPK at fertilization, for example by constitutively activating the upstream MAPK cascade, specifically suppresses cyclin B-cdc2 kinase activation and its consequence, entry into first mitosis. It thus appears that MAPK functions in meiotic maturation by preventing unfertilized eggs from proceeding into parthenogenetic development. The most general effect of artificially maintaining MAPK activity after fertilization is prevention of the G2 to M-phase transition in the first mitotic cell cycle, even though inappropriate reactivation of MAPK after fertilization may lead to metaphase arrest in vertebrates. Advancing the time of MAPK inactivation in fertilized eggs does not, however, speed up their entry into first mitosis. Thus, sustained activity of MAPK during part of the first mitotic cell cycle is not responsible for late entry of fertilized eggs into first mitosis. PMID- 9351825 TI - Cross-cascade activation of ERKs and ternary complex factors by Rho family proteins. AB - Mitogens promote cell growth through integrated signal transduction networks that alter cellular metabolism, gene expression and cytoskeletal organization. Many such signals are propagated through activation of MAP kinase cascades partly regulated by upstream small GTP-binding proteins. Interactions among cascades are suspected but not defined. Here we show that Rho family small G proteins such as Rac1 and Cdc42hs, which activate the JNK/SAPK pathway, cooperate with Raf-1 to activate the ERK pathway. This causes activation of ternary complex factors (TCFs), which regulate c-fos gene expression through the serum response element. Examination of ERK pathway kinases shows that neither MEK1 nor Ras will synergize with Rho-type proteins, and that only MEK1 is fully activated, indicating that MEKs are a focal point for cross-cascade regulation. Rho family proteins utilize PAKs for this effect, as expression of an active PAK1 mutant can substitute for Rho family small G proteins, and expression of an interfering PAK1 mutant blocks Rho-type protein stimulation of ERKs. PAK1 phosphorylates MEK1 on Ser298, a site important for binding of Raf-1 to MEK1 in vivo. Expression of interfering PAK1 also reduces stimulation of TCF function by serum growth factors, while expression of active PAK1 enhances EGF-stimulated MEK1 activity. This demonstrates interaction among MAP kinase pathway elements not previously recognized and suggests an explanation for the cooperative effect of Raf-1 and Rho family proteins on cellular transformation. PMID- 9351824 TI - Regulation of the pp72syk protein tyrosine kinase by platelet integrin alpha IIb beta 3. AB - pp72syk is essential for development and function of several hematopoietic cells, and it becomes activated through tandem SH2 interaction with ITAM motifs in immune response receptors. Since Syk is also activated through integrins, which do not contain ITAMs, a CHO cell model system was used to study Syk activation by the platelet integrin, alpha IIb beta 3. As in platelets, Syk underwent tyrosine phosphorylation and activation during CHO cell adhesion to alpha IIb beta 3 ligands, including fibrinogen. This involved Syk autophosphorylation and the tyrosine kinase activity of Src, and it exhibited two novel features. Firstly, unlike alpha IIb beta 3-mediated activation of pp125FAK, Syk activation could be triggered by the binding of soluble fibrinogen and abolished by truncation of the alpha IIb or beta 3 cytoplasmic tail, and it was resistant to inhibition by cytochalasin D. Secondly, it did not require phosphorylated ITAMs since it was unaffected by disruption of an ITAM-interaction motif in the SH2(C) domain of Syk or by simultaneous overexpression of the tandem SH2 domains. These studies demonstrate that Syk is a proximal component in alpha IIb beta 3 signaling and is regulated as a consequence of intimate functional relationships with the alpha IIb beta 3 cytoplasmic tails and with Src or a closely related kinase. Furthermore, there are fundamental differences in the activation of Syk by alpha IIb beta 3 and immune response receptors, suggesting a unique role for integrins in Syk function. PMID- 9351827 TI - Phosphorylation of activation functions AF-1 and AF-2 of RAR alpha and RAR gamma is indispensable for differentiation of F9 cells upon retinoic acid and cAMP treatment. AB - The role of RAR alpha 1 and RAR gamma 2 AF-1 and AF-2 activation functions and of their phosphorylation was investigated during RA-induced primitive and parietal differentiation of F9 cells. We found that: (i) primitive endodermal differentiation requires RAR gamma 2, whereas parietal endodermal differentiation requires both RAR gamma 2 and RAR alpha 1, and in all cases AF-1 and AF-2 must synergize; (ii) primitive endodermal differentiation requires the proline directed kinase site of RAR gamma 2-AF-1, whereas parietal endodermal differentiation additionally requires that of RAR alpha 1-AF-1; (iii) the cAMP induced parietal endodermal differentiation also requires the protein kinase A site of RAR alpha-AF-2, but not that of RAR gamma; and (iv) the AF-1-AF-2 synergism and AF-1 phosphorylation site requirements for RA-responsive gene induction are promoter context-dependent. Thus, AF-1 and AF-2 of distinct RARs exert specific cellular and molecular functions in a cell-autonomous system mimicking physiological situations, and their phosphorylation by kinases belonging to two main signalling pathways is required to enable RARs to transduce the RA signal during F9 cell differentiation. PMID- 9351826 TI - Biphasic activation of p21ras by endothelin-1 sequentially activates the ERK cascade and phosphatidylinositol 3-kinase. AB - Endothelin-1 (ET-1) induces cell proliferation and differentiation through multiple G-protein-linked signaling systems, including p21ras activation. Whereas p21ras activation and desensitization by receptor tyrosine kinases have been extensively investigated, the kinetics of p21ras activation induced by engagement of G-protein-coupled receptors remains to be fully elucidated. In the present study we show that ET-1 induces a biphasic activation of p21ras in rat glomerular mesangial cells. The first peak of activation of p21ras, at 2-5 min, is mediated by immediate association of phosphorylated Shc with the guanosine exchange factor Sos1 via the adaptor protein Grb2. This initial activation of p21ras results in activation of the extracellular signal-regulated kinase (ERK) cascade. We demonstrate that ET-1 signaling elicits a negative feedback mechanism, modulating p21ras activity through ERK-dependent Sos1 phosphorylation, findings which were confirmed using an adenovirus MEK construct. Subsequent to p21ras and ERK deactivation, Sos1 reverts to the non-phosphorylated condition, enabling it to bind again to the Grb2/Shc complex, which is stabilized by persistent Shc phosphorylation. However, the resulting secondary activation of p21ras at 30 min does not lead to ERK activation, correlating with intensive, ET-1-induced expression of MAP kinase phosphatase-1, but does result in increased p21ras associated phosphatidylinositol 3-kinase activity. Our data provide evidence that ET-1-induced biphasic p21ras activation causes sequential stimulation of divergent downstream signaling pathways. PMID- 9351829 TI - Epstein-Barr virus latent membrane protein-1 triggers AP-1 activity via the c-Jun N-terminal kinase cascade. AB - The Epstein-Barr virus latent membrane protein-1 (LMP-1) is an integral membrane protein which transforms fibroblasts and is essential for EBV-mediated B-cell immortalization. LMP-1 has been shown to trigger cellular NF-kappa B activity which, however, cannot fully explain the oncogenic potential of LMP-1. Here we show that LMP-1 induces the activity of the AP-1 transcription factor, a dimer of Jun/Jun or Jun/Fos proteins. LMP-1 effects on AP-1 are mediated through activation of the c-Jun N-terminal kinase (JNK) cascade, but not the extracellular signal-regulated kinase (Erk) pathway. Consequently, LMP-1 triggers the activity of the c-Jun N-terminal transactivation domain which is known to be activated upon JNK-mediated phosphorylation. Deletion analysis indicates that the 55 C-terminal amino acids of the LMP-1 molecule, but not its TRAF interaction domain, are essential for AP-1 activation. JNK-mediated transcriptional activation of AP-1 is the direct output of LMP-1-triggered signaling, as shown by an inducible LMP-1 mutant. Using a tetracycline-regulated LMP-1 allele, we demonstrate that JNK is also an effector of non-cytotoxic LMP-1 signaling in B cells, the physiological target cells of EBV. In summary, our data reveal a novel effector of LMP-1, the SEK/JNK/c-Jun/AP-1 pathway, which contributes to our understanding of the immortalizing and transforming potential of LMP-1. PMID- 9351828 TI - Conservation of a stress response: human heat shock transcription factors functionally substitute for yeast HSF. AB - Heat shock factors (HSF) are important eukaryotic stress responsive transcription factors which are highly structurally conserved from yeast to mammals. HSFs bind as homotrimers to conserved promoter DNA recognition sites called HSEs. The baker's yeast Saccharomyces cerevisiae possesses a single essential HSF gene, while distinct HSF isoforms have been identified in humans. To ascertain the degree of functional similarity between the yeast and human HSF proteins, human HSF1 and HSF2 were expressed in yeast cells lacking the endogenous HSF gene. We demonstrate that human HSF2, but not HSF1, homotrimerizes and functionally complements the viability defect associated with a deletion of the yeast HSF gene. However, derivatives of hHSF1 that give rise to a trimerized protein, through disruption of a carboxyl- or aminoterminal coiled-coil domain thought to engage in intramolecular interactions that maintain the protein in a monomeric state, functionally substitute for yeast HSF. Surprisingly, hHSF2 expressed in yeast activates target gene transcription in response to thermal stress. Moreover, hHSF1 and hHSF2 exhibit selectivity for transcriptional activation of two distinct yeast heat shock responsive genes, which correlate with previously established mammalian HSF DNA binding preferences in vitro. These results provide new insight into the function of human HSF isoforms, and demonstrate the remarkable functional conservation between yeast and human HSFs, critical transcription factors required for responses to physiological, pharmacological and environmental stresses. PMID- 9351830 TI - Inhibition of NF-kappa-B cellular function via specific targeting of the I-kappa B-ubiquitin ligase. AB - Activation of the transcription factor NF-kappa B is a paradigm for signal transduction through the ubiquitin-proteasome pathway: ubiquitin-dependent degradation of the transcriptional inhibitor I kappa B in response to cell stimulation. A major issue in this context is the nature of the recognition signal and the targeting enzyme involved in the proteolytic process. Here we show that following a stimulus-dependent phosphorylation, and while associated with NF kappa B, I kappa B is targeted by a specific ubiquitin-ligase via direct recognition of the signal-dependent phosphorylation site; phosphopeptides corresponding to this site specifically inhibit ubiquitin conjugation of I kappa B and its subsequent degradation. The ligase recognition signal is functionally conserved between I kappa B alpha and I kappa B beta, and does not involve the nearby ubiquitination site. Microinjection of the inhibitory peptides into stimulated cells abolished NF-kappa B activation in response to TNF alpha and the consequent expression of E-selectin, an NF-kappa B-dependent cell-adhesion molecule. Inhibition of NF-kappa B function by specific blocking of ubiquitin ligase activity provides a novel approach for intervening in cellular processes via regulation of unique proteolytic events. PMID- 9351831 TI - Direct evidence for SIR2 modulation of chromatin structure in yeast rDNA. AB - The yeast SIR2 gene maintains inactive chromatin domains required for transcriptional repression at the silent mating-type loci and telomeres. We previously demonstrated that SIR2 also acts to repress mitotic and meiotic recombination between the tandem ribosomal RNA gene array (rDNA). Here we address whether rDNA chromatin structure is altered by loss of SIR2 function by in vitro and in vivo assays of sensitivity to micrococcal nuclease and dam methyltransferase, respectively, and present the first chromatin study that maps sites of SIR2 action within the rDNA locus. Control studies at the MAT alpha locus also revealed a previously undetected MNase-sensitive site at the a1-alpha 2 divergent promoter which is protected in sir2 mutant cells by the derepressed a1-alpha 2 regulator. In rDNA, SIR2 is required for a more closed chromatin structure in two regions: SRR1, the major SIR-Responsive Region in the non transcribed spacer, and SRR2, in the 18S rRNA coding region. None of the changes in rDNA detected in sir2 mutants are due to the presence of the a1-alpha 2 repressor. Reduced recombination in the rDNA correlates with a small, reproducible transcriptional silencing position effect. Deletion and overexpression studies demonstrate that SIR2, but not SIR1, SIR3 or SIR4, is required for this rDNA position effect. Significantly, rDNA transcriptional silencing and rDNA chromatin accessibility respond to SIR2 dosage, indicating that SIR2 is a limiting component required for chromatin modeling in rDNA. PMID- 9351832 TI - Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells. AB - Genomic reprogramming of primordial germ cells (PGCs), which includes genome-wide demethylation, prevents aberrant epigenetic modifications from being transmitted to subsequent generations. This process also ensures that homologous chromosomes first acquire an identical epigenetic status before an appropriate switch in the imprintable loci in the female and male germ lines. Embryonic germ (EG) cells have a similar epigenotype to PGCs from which they are derived. We used EG cells to investigate the mechanism of epigenetic modifications in the germ line by analysing the effects on a somatic nucleus in the EG-thymic lymphocyte hybrid cells. There were striking changes in methylation of the somatic nucleus, resulting in demethylation of several imprinted and non-imprinted genes. These epigenetic modifications were heritable and affected gene expression as judged by re-activation of the silent maternal allele of Peg1/Mest imprinted gene in the somatic nucleus. This remarkable change in the epigenotype of the somatic nucleus is consistent with the observed pluripotency of the EG-somatic hybrid cells as they differentiated into a variety of tissues in chimeric embryos. The epigenetic modifications observed in EG-somatic cell hybrids in vitro are comparable to the reprogramming events that occur during germ cell development. PMID- 9351833 TI - Dual role for fimbriata in regulating floral homeotic genes and cell division in Antirrhinum. AB - The fimbriata (fim) gene of Antirrhinum affects both the identity and arrangement of organs within the flower, and encodes a protein with an F-box motif. We show that FIM associates with a family of proteins, termed FAPs (FIM-associated proteins), that are closely related to human and yeast Skp1 proteins. These proteins form complexes with F-box-containing partners to promote protein degradation and cell cycle progression. The fap genes are expressed in inflorescence and floral meristems in a pattern that incorporates the domain of fim expression, supporting an in vivo role for a FIM-FAP complex. Analysis of a series of novel fim alleles shows that fim plays a key role in the activation of organ identity genes. In addition, fim acts in the regions between floral organs to specify the correct positioning and maintenance of morphological boundaries. Taking these results together, we propose that FIM-FAP complexes affect both gene expression and cell division, perhaps by promoting selective degradation of regulatory proteins. This may provide a mechanism by which morphological boundaries can be aligned with domains of gene expression during floral development. PMID- 9351834 TI - The asymmetric distribution of the constituents of the Ran system is essential for transport into and out of the nucleus. AB - The GTPase Ran is essential for nuclear import of proteins with a classical nuclear localization signal (NLS). Ran's nucleotide-bound state is determined by the chromatin-bound exchange factor RCC1 generating RanGTP in the nucleus and the cytoplasmic GTPase activating protein RanGAP1 depleting RanGTP from the cytoplasm. This predicts a steep RanGTP concentration gradient across the nuclear envelope. RanGTP binding to importin-beta has previously been shown to release importin-alpha from -beta during NLS import. We show that RanGTP also induces release of the M9 signal from the second identified import receptor, transportin. The role of RanGTP distribution is further studied using three methods to collapse the RanGTP gradient. Nuclear injection of either RanGAP1, the RanGTP binding protein RanBP1 or a Ran mutant that cannot stably bind GTP. These treatments block major export and import pathways across the nuclear envelope. Different export pathways exhibit distinct sensitivities to RanGTP depletion, but all are more readily inhibited than is import of either NLS or M9 proteins, indicating that the block of export is direct rather than a secondary consequence of import inhibition. Surprisingly, nuclear export of several substrates including importin-alpha and -beta, transportin, HIV Rev and tRNA appears to require nuclear RanGTP but may not require GTP hydrolysis by Ran, suggesting that the energy for their nuclear export is supplied by another source. PMID- 9351835 TI - The crystal structure of the human DNA repair endonuclease HAP1 suggests the recognition of extra-helical deoxyribose at DNA abasic sites. AB - The structure of the major human apurinic/ apyrimidinic endonuclease (HAP1) has been solved at 2.2 A resolution. The enzyme consists of two symmetrically related domains of similar topology and has significant structural similarity to both bovine DNase I and its Escherichia coli homologue exonuclease III (EXOIII). A structural comparison of these enzymes reveals three loop regions specific to HAP1 and EXOIII. These loop regions apparently act in DNA abasic site (AP) recognition and cleavage since DNase I, which lacks these loops, correspondingly lacks AP site specificity. The HAP1 structure furthermore suggests a mechanism for AP site binding which involves the recognition of the deoxyribose moiety in an extrahelical conformation, rather than a 'flipped-out' base opposite the AP site. PMID- 9351836 TI - Mechanism of open complex and dual incision formation by human nucleotide excision repair factors. AB - During nucleotide excision repair in human cells, a damaged DNA strand is cleaved by two endonucleases, XPG on the 3' side of the lesion and ERCC1-XPF on the 5' side. These structure-specific enzymes act at junctions between duplex and single stranded DNA. ATP-dependent formation of an open DNA structure of approximately 25 nt around the adduct precedes this dual incision. We investigated the mechanism of open complex formation and find that mutations in XPB or XPD, the DNA helicase subunits of the transcription and repair factor TFIIH, can completely prevent opening and dual incision in cell-free extracts. A deficiency in XPC protein also prevents opening. The absence of RPA, XPA or XPG activities leads to an intermediate level of strand separation. In contrast, XPF or ERCC1 defective extracts open normally and generate a 3' incision, but fail to form the 5' incision. This same repair defect was observed in extracts from human xeroderma pigmentosum cells with an alteration in the C-terminal domain of XPB, suggesting that XPB has an additional role in facilitating 5' incision by ERCC1 XPF nuclease. These data support a mechanism in which TFIIH-associated helicase activity and XPC protein catalyze initial formation of the key open intermediate, with full extension to the cleavage sites promoted by the other core nucleotide excision repair factors. Opening is followed by dual incision, with the 3' cleavage made first. PMID- 9351837 TI - DnaA protein binding to individual DnaA boxes in the Escherichia coli replication origin, oriC. AB - The formation of nucleoprotein complexes between the Escherichia coli initiator protein DnaA and the replication origin oriC was analysed in vitro by band-shift assays and electron microscopy. DnaA protein binds equally well to linear and supercoiled oriC substrates as revealed by analysis of the binding preference to individual DnaA boxes (9-mer repeats) in oriC, and by a competition band-shift assay. DnaA box R4 (oriC positions 260-268) binds DnaA preferentially and in the oriC context with higher affinity than expected from its binding constant. This effect depends on oriC positions 249 to 274, is enhanced by the wild-type sequence in the DnaA box R3 region, but is not dependent on Dam methylation or the curved DNA segment to the right of oriC. DnaA binds randomly to the DnaA boxes R1, M, R2 and R3 in oriC with no apparent cooperativity: the binding preference of DnaA to these sites was not altered for templates with mutated DnaA box R4. In the oriC context, DnaA box R1 binds DnaA with lower affinity than expected from its binding constant, i.e. the affinity is reduced to approximately that of DnaA box R2. Higher protein concentrations were required to observe binding to DnaA box M, making this low-affinity site a novel candidate for a regulatory dnaA box. PMID- 9351838 TI - Intramolecular synapsis of duplex DNA by vaccinia topoisomerase. AB - Complexes formed by vaccinia topoisomerase I on plasmid DNA were visualized by electron microscopy. The enzyme formed intramolecular loop structures in which non-contiguous DNA segments were synapsed within filamentous protein stems. At high enzyme concentrations the DNA appeared to be zipped up within the protein filaments such that the duplex was folded back on itself. Formation of loops and filaments was also observed with an active site mutant, Topo-Phe274. Binding of Topo-Phe274 to relaxed DNA circles in solution introduced torsional strain, which, after relaxation by catalytic amounts of wild-type topo-isomerase, resulted in acquisition of negative supercoils. We surmise that the topoisomerase DNA complex is a plectonemic supercoil in which the two duplexes encompassed by the protein filaments are interwound in a right handed helix. We suggest that topoisomerase-mediated DNA synapsis plays a role in viral recombination and in packaging of the 200 kbp vaccinia genome during virus assembly. PMID- 9351839 TI - Functional differences between the human LINE retrotransposon and retroviral reverse transcriptases for in vivo mRNA reverse transcription. AB - We have analysed the reverse transcriptase (RT) activity of the human LINE retrotransposon and that of two retroviruses, using an in vivo assay within mammalian (murine and human) cells. The assay relies on transfection of the cells with expression vectors for the RT of the corresponding elements and PCR analysis of the DNA extracted 2-4 days post-transfection using primers bracketing the intronic domains of co-transfected reporter genes or of cellular genes. This assay revealed high levels of reverse-transcribed cDNA molecules, with the intron spliced out, with expression vectors for the LINE. Generation of cDNA molecules requires LINE ORF2, whereas ORF1 is dispensable. Deletion derivatives within the 3.8 kb LINE ORF2 allowed further delineation of the RT domain: > 0.7 kb at the 5' end of the LINE ORF2 is dispensable for reverse transcription, consistent with this domain being an endonuclease-like domain, as well as 1 kb at the 3'-end, a putative RNase H domain. Conversely, the RT of the two retroviruses tested, Moloney murine leukemia virus and human immunodeficiency virus, failed to produce similar reverse transcripts. These experiments demonstrate a specific and high efficiency reverse transcription activity for the LINE RT, which applies to RNA with no sequence specificity, including those from cellular genes, and which might therefore be responsible for the endogenous activity that we previously detected within mammalian cells through the formation of pseudogene-like structures. PMID- 9351874 TI - Spinal and femoral DXA for the assessment of spinal osteoporosis. AB - The objective was to determine the diagnostic sensitivity of spinal and femoral dual x-ray absorptiometry (DXA) and to study whether a combination of both sites may enhance discriminatory capability in regard to the presence of vertebral fractures. Spinal and femoral DXA were obtained in 324 postmenopausal women, of whom 90 had at least one vertebral fracture. Age-adjusted logistic regression analyses, ROC analyses, and sensitivity-specificity statistics were used to assess the discriminatory ability of spinal and femoral bone density (BMD) alone and in combination. The age-adjusted odds ratios per standard deviation decrease in BMD (OR) for spinal and femoral measurements were comparable (Ward's triangle: OR = 1.62; femoral neck: OR = 1.51; total hip: OR = 1.47; spine: OR = 1.34). Combining spinal and femoral bone density measurements did not improve diagnostic sensitivity of DXA considerably as compared to using BMD of a single site and adjusting the "fracture threshold." The conclusion drawn is that spinal and femoral BMD measurements using DXA have a comparable diagnostic sensitivity for vertebral fracture discrimination. Different individuals at risk for osteoporosis may be identified using both methods. The clinical usefulness of a combination of two bone density measurements needs further study in a prospective setting. PMID- 9351875 TI - Comparison of three bone densitometry methods in osteoporotic women. AB - Three techniques of bone mass measurement were evaluated in the diagnosis of postmenopausal osteoporosis; the overlap in the measurements and the capacity for discriminating was determined among 51 postmenopausal normal (mean age 66.6 +/- 8.4 years) and 42 postmenopausal osteoporotic women (mean age 68.5 +/- 7.5 years). All bone mass was evaluated by total body bone mineral content (BMCTB), density (BMDTB), ultrasound bone velocity (UBV) in proximal phalanxes 2-5 of the nondominant hand (UBV = mean value of all ultrasound measurements), and peripheral quantitative computed tomography of the nondominant forearm (pQCT). BMCTB was found to be significantly better (P < 0.0001) for diagnosing postmenopausal osteoporosis than the other methods; both cortical and trabecular pQCT measurements were more discriminating than the corresponding UBV measurements (P < 0.001). T-score values in normals, subjects versus osteoporotic ones were BMCTB -1.15 +/- 0.79 versus -3.17 +/- 0.74; BMDTB -1.01 +/- 0.97 versus -3.28 +/- 0.81; UBV -1.51 +/- 1.02 versus -2.34 +/- 1.21; trabecular-pQCT -0.40 +/- 0.72 versus -1.57 +/- 0.37; cortical-pQCT -1.00 +/- 0.87 versus -2.67 +/- 0.53; and total-pQCT -0.65 +/- 1.01 versus -2.34 +/- 0.27, respectively. The overlap in values between the postmenopausal normal and postmenopausal osteoporotic groups was 50% with UBV, 6% with BMCTB, 9% with BMDTB, 25% with cortical pQCT, and 42% with trabecular pQCT. BMCTB, BMDTB, UBV, and pQCT correlated well with each other as measurements of bone mass, but BMCTB was more discriminating than the other measurements in the diagnosis of osteoporosis. PMID- 9351840 TI - Plus-strand strong-stop DNA transfer in yeast Ty retrotransposons. AB - The yeast Ty1 LTR retrotransposon replicates by reverse transcription and integration; the process shows many similarities to the retroviral life cycle. However, we show that plus strand strong-stop DNA transfer in yeast Ty1 elements differs from the analogous retroviral process. By analysis of the native structure of the Ty1 primer binding site and by a series of manipulations of this region and assessment of the effects on retrotransposition, we show that primer binding site inheritance is not from the tRNA primer, which is inconsistent with classical retroviral models. This unusual inheritance pattern holds even when the Ty1 primer binding site is lengthened in order to be more retrovirus-like. Finally, the distantly related Ty3 element has an inheritance pattern like Ty1, indicating evolutionary conservation of the alternative pathway used by Ty1. Based on these results we arrive at a plus strand primer recycling model that explains Ty1 plus strand strong-stop DNA transfer and inheritance patterns in the primer binding site. PMID- 9351872 TI - Truncation of the cytoplasmic domain of beta3 in a variant form of Glanzmann thrombasthenia abrogates signaling through the integrin alpha(IIb)beta3 complex. AB - Glanzmann thrombasthenia is an inherited bleeding disorder characterized by absence or dysfunction of the platelet integrin alpha(IIb)beta3. Patient RM is a thrombasthenic variant whose platelets fail to aggregate in response to physiological agonists, despite the fact that they express abundant levels of alpha(IIb)beta3 on their surface. Binding of soluble fibrinogen or fibrinogen mimetic antibodies to RM platelets did not occur, except in the presence of ligand-induced binding site (LIBS) antibodies that transformed the RM integrin complex into an active conformation from outside the cell. Sequence analysis of PCR-amplified genomic DNA and platelet mRNA revealed a C2268T nucleotide substitution in the gene encoding the integrin beta3 subunit that resulted in an Arg724Ter mutation, producing a truncated protein containing only the first eight of the 47 amino acids normally present in the cytoplasmic domain. Functional analysis of both RM platelets and CHO cells stably expressing this truncated integrin revealed that the alpha(IIb)beta3Arg724Ter complex is able to mediate binding to immobilized fibrinogen, though downstream events, including cytoskeletally-mediated cell spreading and tyrosine phosphorylation of focal adhesion kinase, pp125FAK, fail to occur. These studies establish the importance of the membrane-distal portion of the integrin beta3 cytoplasmic domain in bidirectional transmembrane signaling in human platelets, and the role of integrin signaling in maintaining normal hemostasis in vivo. PMID- 9351876 TI - Comparison of assay of total and bone-specific alkaline phosphatase in the assessment of osteoblast activity in patients with metastatic bone disease. AB - The evaluation of response of osseous metastases to systemic treatments is often low as a consequence of the different radiologic appearances that make objective assessment not only difficult but sometimes impossible. Radiographic evidence of recalcification, the UICC criterion of response, is often evident for 6 months and sometimes may be delayed even more. This accounts for lower response rates in bone with respect to other metastatic sites in clinical trials. A transient rise in bone formation indices may provide an early indication of bone healing and, along with measurement of symptomatic changes, could ameliorate the response evaluation. Among the biochemical markers of bone formation, total alkaline phosphatase (TALP) is widely employed, but it lacks specificity. Estimation of bone isoenzyme (E-BALP) by electrophoretic techniques is time consuming and semiquantitative. The immunoradiometric assay (I-BALP) seems to overcome these limitations. In this study, we compared the two methods of bone isoenzyme estimation with each other and with the levels of bone gla protein (BGP) and carboxyterminal propeptide of type I procollagen (PICP) in a group of 136 cancer patients with bone metastases stratified as having lytic or mixed and blastic lesions at X-ray, and in 62 cancer patients without apparent bone involvement. The same indices were also evaluated prospectively in a patient subset submitted to chemotherapy associated with pamidronate. The aims of the study were to evaluate whether I-BALP is superior to E-BALP and whether both methods of bone isoenzyme estimation are more advantageous than TALP, BGP, and PICP in the assessment of osteoblast activity either in baseline conditions or in response to treatment. In bone metastatic patients with lytic appearances, values above the cut-off limit were observed in 32.1%, 23.3%, 48.9%, 32.9%, and 14% for, TALP, E BALP, I-BALP, PICP, and BGP, while the corresponding percentages in those with blastic/mixed appearances were 74.0%, 84.8%, 76.9%, 51.9%, and 43.8%, respectively. In the patients without bone involvement, values within the normal range were 90.2%, 98.2%, 89.6%, 71.7%, and 90.2%, respectively. Levels of TALP, E BALP, and I-BALP were reciprocally correlated in the three groups examined. In bone metastatic patients, however, the degree of correlation of the enzymes with PICP and BGP was weak. Liver isoenzyme of alkaline phosphatase (LALP) was found to correlate with E-BALP, but not with I-BALP, in patients with mixed/blastic lesions. Thirty-eight patients were submitted to pamidronate therapy (60 mg every 3 weeks, administered 4 times) in association with cytotoxic treatment. Osteoblastic markers were determined before any administration. Serum TALP, E BALP, and I-BALP showed a transient rise in 9 cases, a progressive reduction in 12, no change in 2, and a progressive increase in 6. Changes in E-BALP and I-BALP from baseline were greater than those of TALP. A divergent pattern between TALP and both I-BALP and E-BALP was found in 9 patients, whereas a divergent temporal profile between the two methods of bone isoenzyme estimation was recorded in only 3 patients. Eight out of 38 cases obtained a partial recalcification of lytic and mixed lesions. Seven of them showed the concomitant early increase in TALP, E BALP, and I-BALP followed by a gradual decline (osteoblastic flare), whereas 1 patient demonstrated the flare of E-BALP and I-BALP but not of TALP. No relationship was found between response and temporal changes in in BGP and PICP serum levels. We conclude that I-BALP is a useful marker for detecting excess osteoblastic activity in patients who have at imaging "pure" lytic bone metastases. In the longitudinal evaluation of patients receiving multiple pamidronate infusions plus chemotherapy, TALP, E-BALP, and I-BALP, but not BGP and PICP, appeared to be useful to identify responders in bone. (ABSTRACT TRUNCATED) PMID- 9351877 TI - Identification of metabolic bone disease in patients with endogenous hyperthyroidism: role of biological markers of bone turnover. AB - Active hyperthyroidism is associated with reduced bone mass. Nevertheless, not all patients show the same risk for developing osteoporosis. Our aim was to analyze some clinical and biochemical potential predictors of low bone mass in hyperthyroid patients. We studied 127 consecutive hyperthyroid patients (110 females, 17 males; aged 42 +/- 16 years). Bone mineral density (BMD) was measured by dual X-ray absorptiometry (DXA) at lumbar spine (LS; L2-L4) and femoral neck (FN). Data were expressed as g/cm2 and T-score. Patients were placed into two groups based on recent WHO criteria: Group A, no osteoporosis (n = 98); and group B, lumbar or femoral osteoporosis (n = 29). Study protocol included evaluation of osteoporosis risk factors, anthropometrical variables, thyroid function, and bone turnover markers. Receiver-operating characteristic (ROC) plots for the precision of bone markers and multivariate analysis for the prediction of BMD and osteoporosis were performed. Group B showed greater age and proportion of menopausal females; lower weight, height, and calcium intake; longer duration of menopause; and greater levels of total and bone alkaline phosphatase and of urine hydroxyproline. No differences in thyroid function, osteocalcin, tartrate resistant acid phosphatase, and type I collagen C-telopeptide (ICTP) were found. The best predictive model accounted for 46% and 62% of the variability of lumbar and femoral BMD respectively and correctly classified 89% of the osteoporotic hyperthyroid patients. No significant difference in ROC plots was observed. It is concluded that hyperthyroid patients with lumbar or femoral osteoporosis show a typical clinical and biochemical profile illustrating that the relationship between BMD and bone markers is better in high turnover states. Classical bone turnover markers show high performance in the evaluation of hyperthyroid bone disease. PMID- 9351878 TI - Not all postmenopausal women on chronic steroid and estrogen treatment are osteoporotic: predictors of bone mineral density. AB - Chronic steroid use results in osteoporosis, and postmenopausal women are believed to be at a high risk for steroid-induced bone loss. The purpose of this study was to determine predictors of bone mineral density (BMD) in postmenopausal women on both chronic steroid and hormone replacement therapy. Seventy-six postmenopausal women (> or = 3 years postmenopausal, > or = 2 years of steroid treatment of > or = 5 mg/day of prednisone, and > or 1 year of hormone replacement therapy) were recruited into this study. Measurements of BMD of the lumbar spine and femoral neck were obtained in all subjects. Risk factors for osteoporosis were obtained by questionnaire. Discriminant analysis was performed to determine predictors of BMD. Osteoporosis, defined by a T score of < -2.5, was present in the lumbar spine or femoral neck in 34 of the 76 subjects. Based on these criteria, women with osteoporosis were significantly older, were more years postmenopausal, and had a lower body mass index (BMI) than women who did not have osteoporosis. Predictors of osteoporosis for both the femoral neck and spine included a low BMI (P < 0.05), more years postmenopausal (P < 0.01), and more years on steroids (P < 0.01). Low BMI was the only significant predictor of osteoporosis in the lumbar spine (P < 0.05), whereas for the femoral neck both years on steroids (P < 0.05) and BMI (P < 0.05) were significant predictors of low BMD. In summary, not all postmenopausal women on chronic steroid and hormone replacement therapy are osteoporotic but a low BMI, more years on steroids, and more years postmenopausal were significant predictors of osteoporosis in these subjects. PMID- 9351879 TI - Prevention of corticosteroid-induced osteoporosis with alendronate in sarcoid patients. AB - Prolonged corticosteroid administration, as often required in the treatment of sarcoidosis, increases the risk of osteoporosis and fracture. The aim of the present study was to evaluate the usefulness of alendronate, a third generation bisphosphonate, in preventing corticosteroid-induced osteoporosis. Forty-three consecutive, previously untreated, sarcoid patients (17 men and 26 premenopausal women) were included in the study: 13 needed no treatment and served as controls (Group 1) and 30 needed glucocorticoids (prednisone) and were randomly selected to also receive either placebo (n = 15, Group 2) or alendronate 5 mg/day (n = 15, Group 3). Bone mineral density (BMD) at the ultradistal radius by dual photon absorptiometry (Osteograph 1000, NIM, Verona, Italy) and biochemical markers of bone turnover were measured at baseline and after 6 and 12 months of glucocorticoid therapy. No significant difference was found between Groups 2 and 3 in the mean cumulative dose of prednisone (4945 +/- 1956 mg and 5110 +/- 2013 mg, respectively). At the end of the study period, BMD increased by 0.8% in the alendronate-treated group; in the placebo-treated group, BMD decreased by 4.5%. The difference between groups was significant (P < 0.01, ANOVA). A significant decrease in markers of bone formation was found in all patients treated with prednisone (Groups 2 and 3), independently of alendronate. Alendronate, however, counteracted the increase in markers of bone resorption induced by glucocorticoid therapy. Our data suggest that alendronate is effective in preventing glucocorticoid-induced bone loss in sarcoid patients. Further studies on alendronate use in steroid-induced osteoporosis are needed. PMID- 9351881 TI - The effect of weight change on DXA scans in a 2-year trial of etidronate therapy. AB - Variation in soft tissue composition is a potential cause of error in dual X-ray absorptiometry (DXA) measurements of bone mineral density (BMD). We investigated the effect of patients' change of weight on DXA scans in 152 women enrolled in a 2-year trial of cyclical etidronate therapy. Scans of the spine, hip, and total body were performed at baseline, 1 and 2 years on a Hologic QDR-2000. The study was completed by 135 subjects (64 on etidronate, 71 on placebo). Results were expressed as the percentage change in BMD (spine, femoral neck, total body) or bone mineral content (BMC)(total body only) at 2 years. Total body scans were analyzed using the manufacturer's 'standard' and 'enhanced' algorithms. Analysis was performed using multivariate regression with percentage change in BMD or BMC as the dependent variable, and treatment group and percentage change in weight as the independent variables. Weight change varied between -14.4% and +16.7%. All DXA variables showed a statistically significant treatment effect. Standard total body BMD and BMC and enhanced total body BMC all showed a significant dependence on weight change (P < 0.01, P < 0.001 and P < 0.01, respectively). No effect of weight change was seen on spine, femoral neck, or enhanced total body BMD. In order to investigate the effects of weight on long-term precision, patients were allocated to two groups according to baseline body mass index (BMI < 25 and > 25 kg/m2, respectively). For femoral neck BMD the root mean square (RMS) residual percentage change was statistically significantly larger in the high BMI group (P < 0.05) but all other bone density variables showed no significant difference. With patients allocated to two groups according to their absolute percentage change in weight (< 5% and > 5%, respectively) the RMS residual percentage changes in the bone density variables were statistically significantly larger in the large weight change group for femoral neck BMD (P < 0.05) and for standard and enhanced total body BMC (P < 0.01 and P < 0.05, respectively). With the exception of the standard total body algorithm, weight change in a longitudinal study of postmenopausal women was not found to cause systematic errors in the results of DXA studies but may adversely affect precision. PMID- 9351882 TI - TSC-36 (follistatin-related polypeptide) gene expression in estrogen receptor positive osteoblastic cell line, CDO7F. AB - Using an osteoblastic cell line (CDO7F) that was selected as an estrogen receptor positive cell by immunocytochemical detection and reverse transcription polymerase chain reaction (RT-PCR) subtractive cDNA cloning using oligo(dT)30 Latex and PCR was performed to isolate a gene that is induced by estrogen. A cDNA clone was isolated and its nucleotide sequence identified it as TSC 36, previously called follistatin-related polypeptide. Northern blot analysis showed that the TSC-36 gene expression was certainly upregulated by estrogen. Tamoxifen also upregulated this gene expression. These findings indicate that TSC-36 may be one of the estrogen-regulated genes in osteoblastic cells. PMID- 9351880 TI - An in vitro and in vivo study of cytokines in the acute-phase response associated with bisphosphonates. AB - We studied the acute phase response, including specific cytokine production, [interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor alpha(TNF alpha)] following a single dose of Aredia (disodium pamidronate) in patients with increased bone turnover and, in vitro, the role played by specific cytokines in the acute-phase reaction which may follow the administration of aminobisphosphonates. An in vivo exploratory study was done on 24 in- and outpatients with increased bone turnover given a single intravenous dose of pamidronate 60 mg. Measurements were taken at baseline and at 24, 48, and 72 hours. The main outcome measures were changes from baseline in serum IL-1, IL-6, and TNF alpha. In addition, C-reactive protein (CRP), white blood cell count (WCC), lymphocyte count, and elastase concentration were measured. Symptomatic evaluation was made of fever, bone pain, and rigors. In vitro, whole blood from eight healthy volunteers was exposed to various concentrations of the three bisphosphonates--pamidronate, clodronate, and zoledronate. Measurements were taken immediately before and at 3, 6, and 10 hours after exposure to drugs. The main outcome measures were changes in serum IL-1, IL-6, and TNF alpha. In vivo, there was a statistically significant (P < 0.001) increase in median values of TNF alpha in all post-baseline measurements. Median values for IL-6 also showed a significant (P < 0.001) increase at 24 hours after dosing. There were no statistically significant changes in median IL-1 values. Few patients showed any change from baseline in total WCC or in lymphocyte count, but 62.5% of patients with normal range baseline values for CRP increased to above normal levels after treatment. Fourteen patients experienced fever; 2 reported rigors. There was no correlation between fever and changes in cytokines. There were no serious adverse experiences or premature discontinuations due to poor tolerability, and 91% of the patients expressed willingness to receive pamidronate again. In vitro, an increase in TNF alpha and a mild increase in IL-6 was seen with all bisphosphonates, with the greatest effects seen with the highest concentration of both pamidronate and zoledronate. No changes were observed in IL-1 with any agent. Significant changes in both TNF alpha and IL-6 were observed within 3 days of a single dose of pamidronate in patients treated for the first time confirming previous findings. However, the lack of change in IL-1 in vivo and in vitro does not support the hypothesis that this cytokine plays a major role in the acute phase reaction. The cellular mechanism of the interaction among aminobisphosphonates, IL-6, and TNF alpha requires further investigations. The results of the in vitro study are consistent with the in vivo findings. PMID- 9351883 TI - Triiodothyronine, a regulator of osteoblastic differentiation: depression of histone H4, attenuation of c-fos/c-jun, and induction of osteocalcin expression. AB - Thyroid hormones influence growth and differentiation of bone cells. In vivo and in vitro data indicate their importance for development and maintenance of the skeleton. Triiodothyronine (T3) inhibits proliferation and accelerates differentiation of osteoblasts. We studied the regulatory effect of T3 on markers of proliferation as well as on specific markers of the osteoblastic phenotype in cultured MC3T3-E1 cells at different time points. In parallel to the inhibitory effect on proliferation, T3 down-regulated histone H4 mRNA expression. Early genes (c-fos/c-jun) are highly expressed in proliferating cells and are down regulated when the cells switch to differentiation. When MC3T3-E1 cells are cultured under serum-free conditions, basal c-fos/c-jun expressions are nearly undetectable. Under these conditions, c-fos/c-jun mRNAs can be stimulated by EGF, the effect of which is attenuated to about 46% by T3. In addition, T3 stimulated the expression at the mRNA and protein level of osteocalcin, a marker of mature osteoblasts and alkaline phosphatase activity. All these effects were more pronounced when cells were cultured for more than 6 days. These data indicate that T3 acts as a differentiation factor in osteoblasts by influencing the expression of cell cycle-regulated, of cell growth-regulated, and of phenotypic genes. PMID- 9351884 TI - Analysis of whole skeleton 3H-tetracycline loss as a measure of bone resorption in maturing rats. AB - Several modifications of the 3H-tetracycline bone labeling method for measuring whole skeleton bone resorption were tested. Under steady state conditions of whole skeleton resorptive activity, bone labeling for intervals longer than 2 weeks prior to experimentation did not significantly alter the urinary 3H tetracycline loss curve. The utilization of nonlinear regression analysis showed that the urinary loss of 3H-tetracycline was best described by double exponential equations, indicating the loss of label from two distinct and independent exchangeable bone compartments. This conclusion was supported by the finding that soft tissues were effectively depleted of 3H-tetracycline by 24 hours after the final injection of label. Hence, it was concluded that approximately 40% of the 3H-tetracycline loss from skeletal bone is associated with a "fast" compartment which is depleted within 6 or 7 days after label loading. The size and rate of 3H tetracycline loss from the fast compartment decreased (40%) with age such that the depletion time remained constant between 8 and 24 weeks of age in both male and female rats. The remaining 60% of 3H-tetracycline loss from a "slow" compartment which was depleted in about 70 days in young (8 week) rats. This compartment, which is believed to reflect cell-mediated resorption of calcified bone; decreased in size with age in both male (50%) and female (30%) rats. The rate of label loss from this compartment, however, remained relatively high so that the depletion time decreased (approximately 35%) between 8 and 24 weeks of age. By determining whole skeletal mass and calculating these parameters on the basis of skeletal mass, we were further able to demonstrate significantly higher resorptive activity in female than in male rats by 24 weeks of age. PMID- 9351885 TI - The domain of hypertrophic chondrocytes in growth plates growing at different rates. AB - In this study, we tested the hypotheses that (a) both the domain volume (volume of the cell and the matrix it has formed) and matrix volume of juxtametaphyseal hypertrophic chondrocytes in the growth plate is tightly controlled, and that (b) the domain volume of juxtametaphyseal hypertrophic chondrocytes is a strong determinant of the rate of bone length growth. We analyzed the rate of bone length growth (oxytetracycline labeling techniques) and nine stereologic and kinetic parameters related to the juxtametaphyseal chondrocytic domain in the proximal and distal radial and tibial growth plates of 21- and 35-day-old rats. The domain volume increased with increasing growth rates, independent of the location of the growth plate and the age of the animal. Within age groups, the matrix volume per cell increased with increasing growth rates, but an identical growth plate had the same matrix volume per cell in 21- and 35-day-old rats. The most suitable regression model (R2 = 0.992) to describe the rate of bone length growth included the mean volume of juxtametaphyseal hypertrophic chondrocytes and the mean rate of cell loss/cell proliferation. This relationship was independent of the location of the growth plate and the age of the animal. The data suggest that the domain volume of juxtametaphyseal hypertrophic chondrocytes, as well as the matrix volume produced per cell, may be tightly regulated. In addition, the volume of juxtametaphyseal hypertrophic chondrocytes and the rate of cell loss/rate of cell proliferation may play the most important role in the determination of the rate of bone length growth. PMID- 9351887 TI - Molecular properties of hepatic uptake systems for bile acids and organic anions. PMID- 9351886 TI - PTHrP(107-111) inhibits in vivo resorption that was stimulated by PTHrP(1-34) when applied intermittently to neonatal mice. AB - Stimulated resorption by PTH(1-34) and PTHrP(1-34) was studied in a neonatal mouse model by injecting a relatively high dose (0.2 microgram/g BW) of the peptides for 6 or 16 consecutive daily injections. 3H-Tetracycline was applied once, before the beginning of the injection period. Only PTHrP(1-34) increased resorption as revealed by decreased radioactivity remaining in the tibia when compared with vehicle-treated mice. This activity was related to increased numbers of mature osteoclasts in the metaphysis of the young bones. PTHrP(107 111) completely aborted the stimulated resorption when applied simultaneously with PTHrP(1-34). PMID- 9351888 TI - Electrophysiological characterization of the flounder type II Na+/Pi cotransporter (NaPi-5) expressed in Xenopus laevis oocytes. AB - The two electrode voltage clamp technique was used to investigate the steady state and presteady-state kinetic properties of the type II Na+/Pi cotransporter NaPi-5, cloned from the kidney of winter flounder (Pseudopleuronectes americanus) and expressed in Xenopus laevis oocytes. Steady-state Pi-induced currents had a voltage-independent apparent K(m) for Pi of 0.03 mM and a Hill coefficient of 1.0 at neutral pH, when superfusing with 96 mM Na+. The apparent K(m) for Na+ at 1 mM Pi was strongly voltage dependent (increasing from 32 mM at -70 mV to 77 mM at 30 mV) and the Hill coefficient was between 1 and 2, indicating cooperative binding of more than one Na+ ion. The maximum steady-state current was pH dependent, diminishing by 50% or more for a change from pH 7.8 to pH 6.3. Voltage jumps elicited presteady-state relaxations in the presence of 96 mM Na+ which were suppressed at saturating Pi (1 mM). Relaxations were absent in non-injected oocytes. Charge was balanced for equal positive and negative steps, saturated at extremes of potential and reversed at the holding potential. Fitting the charge transfer to a Boltzmann relationship typically gave a midpoint voltage (V0.5) close to zero and an apparent valency of approximately 0.6. The maximum steady state transport rate correlated linearly with the maximum Pi-suppressed charge movement, indicating that the relaxations were NaPi-5-specific. The apparent transporter turnover was estimated as 35 sec-1. The voltage dependence of the relaxations was Pi-independent, whereas changes in Na+ shifted V0.5 to -60 mV at 25 mM Na+. Protons suppressed relaxations but contributed to no detectable charge movement in zero external Na+. The voltage dependent presteady-state behavior of NaPi-5 could be described by a 3 state model in which the partial reactions involving reorientation of the unloaded carrier and binding of Na+ contribute to transmembrane charge movement. PMID- 9351889 TI - Osmotic regulation of Na+ transport across A6 epithelium: interactions with prostaglandin E2 and cyclic AMP. AB - Previous work from this laboratory has shown that apical membrane sodium channel activity is stimulated by serosal hyposmotic solutions (Wills, Millinoff & Crowe, 1991). In the present study, we determined whether this stimulation of sodium transport is additive with the actions of prostaglandin E2 (PGE2) or cyclic AMP (cAMP). Addition of exogenous PGE2 (100 nM; serosal bath) to isosmotic solutions led to large increases in the amiloride-sensitive short-circuit current (Isc) and transepithelial conductance (Gt), whereas no significant effects of PGE2 were observed in hyposmotic serosal solutions. Subsequent addition of mucosal amiloride reduced Isc by approximately 95% and Gt by approximately 60%. Inhibition of endogenous PGE2 production by blockers of phospholipase A2 activity (quinacrine or 3[4-octadecyl]-benzoylacrylic acid; OBBA), or inhibition of cyclooxygenase activity by indomethacin reduced the stimulation of Isc and Gt by hyposmotic solutions. Addition of forskolin (FSK) or 3-Isobutyl-1-methylxanthine (IBMX) also resulted in approximately twofold increases in the amiloride sensitive Isc and Gt and abolished the effects of subsequent hyposmotic challenge. The effects of forskolin, PGE2, and hyposmotic challenge were diminished by pretreatment with H89, a protein kinase A (PKA) inhibitor. We conclude that osmotic regulation of sodium channel activity interacts with multiple intracellular signaling pathways, specifically the arachidonic acid metabolic pathway and the cAMP/PKA intracellular messenger cascade. PMID- 9351890 TI - G-protein regulation of an L-type calcium channel current in canine jejunal circular smooth muscle. AB - Calcium entry into smooth muscle cells is essential to maintain contractility. In canine jejunal circular smooth muscle cells the predominant calcium entry pathway is through L-type calcium channels. The aim of this study was to determine the G protein regulation of L-type calcium channel current (ICaL) in isolated canine jejunal circular smooth muscle cells. Barium (80 mM) was used as the charge carrier. GTP-gamma S and GTP increased maximal inward current from 118.7 +/- 12 pA to 227.5 +/- 21.5 pA (n = 8) and 174.6 +/- 10.1 pA (n = 6) respectively. The increase in inward current was blocked by nifedipine suggesting it was through L type calcium channels. Pertussis toxin did not alter baseline ICaL while cholera toxin increased ICaL from 125 +/- 19 pA in controls (n = 6) to 347 +/- 30 pA (n = 4). Staurosporine inhibited the increase in current evoked by GTP-gamma S and calyculin further increased ICaL over the increase evoked by GTP-gamma S. The results suggest that cholera toxin sensitive G-proteins activate L-type calcium channels in isolated canine jejunal circular smooth muscle cells through protein phosphorylation. PMID- 9351891 TI - Cell swelling activates phospholipase A2 in Ehrlich ascites tumor cells. AB - Ehrlich ascites tumor cells, loaded with 3H-labeled arachidonic acid and 14C labeled stearic acid for two hours, were washed and transferred to either isotonic or hypotonic media containing BSA to scavenge the labeled fatty acids released from the cells. During the first two minutes of hypo-osmotic exposure the rate of 3H-labeled arachidonic acid release is 3.3 times higher than that observed at normal osmolality. Cell swelling also causes an increase in the production of 14C-stearic acid-labeled lysophosphatidylcholine. This indicates that a phospholipase A2 is activated by cell swelling in the Ehrlich cells. Within the same time frame there is no swelling-induced increase in 14C-labeled stearic acid release nor in the synthesis of phosphatidyl 14C-butanol in the presence of 14C-butanol. Furthermore, U7312, an inhibitor of phospholipase C, does not affect the swelling induced release of 14C-labeled arachidonic acid. Taken together these results exclude involvement of phospholipase A1, C and D in the swelling-induced liberation of arachidonic acid. The swelling-induced release of 3H-labeled arachidonic acid from Ehrlich cells as well as the volume regulatory response are inhibited after preincubation with GDP beta S or with AACOCF3, an inhibitor of the 85 kDa, cytosolic phospholipase A2. Based on these results we propose that cell swelling activates a phospholipase A2--perhaps the cytosolic 85 kDa type--by a partly G-protein coupled process, and that this activation is essential for the subsequent volume regulatory response. PMID- 9351892 TI - High-affinity NO(3-)-H+ cotransport in the fungus Neurospora: induction and control by pH and membrane voltage. AB - High-affinity nitrate transport was examined in intact hyphae of Neurospora crassa using electrophysiological recordings to characterize the response of the plasma membrane to NO3- challenge and to quantify transport activity. The NO3(-) associated membrane current was determined using a three electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in hyphae transferred to NO3(-)-free, N-limited medium for 15 hr, and in hyphae grown in the absence of a nitrogen source after a single 2-min exposure to 100 microM NO3-. In the latter, induction showed a latency of 40-80 min and rose in scalar fashion with full transport activity measurable approx. 100 min after first exposure to NO3-; it was marked by the appearance of a pronounced sensitivity of membrane voltage to extracellular NO3- additions which, after induction, resulted in reversible membrane depolarizations of (+)54-85 mV in the presence of 50 microM NO3-; and it was suppressed when NH4+ was present during the first, inductive exposure to NO3-. Voltage clamp measurements carried out immediately before and following NO3- additions showed that the NO3(-)-evoked depolarizations were the consequence of an inward-directed current that appeared in parallel with the depolarizations across the entire range of accessible voltages (-400 to +100 mV). Measurements of NO3- uptake using NO3(-)-selective macroelectrodes indicated a charge stoichiometry for NO3- transport of 1(+):1(NO3 ) with common K(m) and Jmax values around 25 microM and 75 pmol NO3- cm-2sec-1, respectively, and combined measurements of pHo and [NO3-]o showed a net uptake of approx. 1 H+ with each NO3- anion. Analysis of the NO3- current demonstrated a pronounced voltage sensitivity within the normal physiological range between -300 and -100 mV as well as interactions between the kinetic parameters of membrane voltage, pHo and [NO3-]o. Increasing the bathing pH from 5.5 to 8.0 reduced the current and the associated membrane depolarizations 2- to 4-fold. At a constant pHo of 6.1, driving the membrane voltage from -350 to -150 mV resulted in an approx. 3-fold reduction in the maximum current and a 5-fold rise in the apparent affinity for NO3-. By contrast, the same depolarization effected an approx. 20% fall in the K(m) for transport as a function in [H+]o. These, and additional results are consistent with a charge-coupling stoichiometry of 2(H+) per NO3- anion transported across the membrane, and implicate a carrier cycle in which NO3 binding is kinetically adjacent to the rate-limiting step of membrane charge transit. The data concur with previous studies demonstrating a pronounced voltage dependence to high-affinity NO3- transport system in Arabidopsis, and underline the importance of voltage as a kinetic factor controlling NO3- transport; finally, they distinguish metabolite repression of NO3- transport induction from its sensitivity to metabolic blockade and competition with the uptake of other substrates that draw on membrane voltage as a kinetic substrate. PMID- 9351893 TI - Activation of divalent cation influx into S. cerevisiae cells by hypotonic downshift. AB - Subjecting Saccharomyces cerevisiae cells to a hypotonic downshift by transferring cells form YPD medium containing 0.8 M sorbitol to YPD medium without sorbitol induces a transient rapid influx of Ca2+ and other divalent cations into the cell. For cells grown in YPD at 37 degrees C, this hypotonic downshift increases Ca2+ accumulation 6.7-fold. Hypotonic downshift-induced Ca2+ accumulation and steady-state Ca2+ accumulation in isotonic YPD medium are differentially affected by dodecylamine and Mg2+. The Ca(2+)-influx pathway responsible for hypotonic-induced Ca2+ influx may account for about 10-35% of Ca2+ accumulation by cells growing in YPD. Ca2+ influx is not required for cells to survive a hypotonic downshift. Hypotonic downshift greatly reduces the ability of S. cerevisiae cells to survive a 5-min exposure to 10 mM Cd2+ suggesting that mutants resistant to acute Cd2+ exposure may help identify genes required for hypotonic downshift-induced divalent cation influx. PMID- 9351894 TI - Na+/H+ exchanger isoform 2 (NHE2) is expressed in the apical membrane of the medullary thick ascending limb. AB - Apical Na+/H+ exchangers (NHE) in the proximal tubule and medullary thick ascending limb (MTAL) display similar functions and regulation, suggesting that similar NHE isoforms are present. In the rat proximal tubule, NHE2 and NHE3 are present in the apical membrane, however, in the MTAL, NHE3, but not NHE2, mRNA has been found. In this study, the expression and subcellular localization of NHE2 in both rat and mouse MTAL were studied. To detect NHE2 mRNA, reverse transcription-polymerase chain reaction (RT-PCR) was performed in microdissected MTAL tubules using primers specific for NHE2. Analysis of PCR products with and without digestion by restriction enzymes chosen from the published NHE2 sequence gave predicted sizes. Subcloning and sequencing of the PCR product from mouse MTAL revealed 91% and 75% identity to the published NHE2 nucleotide sequence of comparable regions in rat and rabbit, respectively. Thus, NHE2 mRNA is expressed in the MTAL of mouse and rat. The subcellular localization of NHE2 was determined by immunochemistry using a specific NHE2 antibody. Immunofluorescence staining was observed in the apical, but not basolateral, membrane of MTAL of both species. In addition, anti-NHE2 antibody recognized an 85 kD protein in plasma membranes prepared from mouse and rat renal outer medulla and a MTAL cell line by Western analysis, which further support that NHE2 protein is expressed in the MTAL of both species. We conclude that NHE2 is expressed in the apical membrane of MTAL in both mouse and rat. PMID- 9351895 TI - A p53 growth arrest protects fibroblasts from anticancer agents. AB - Reversible inhibitors of the cell cycle such as the TGF-betas have been exploited to protect dividing cells from exposure to anticancer drugs and radiation. Here, rat embryo fibroblast (REF) lines expressing different p53 mutations were used to test whether the p53 growth arrest could also chemoprotect cells from high doses of anticancer drugs. Whereas the doubling times of the different REF lines at 37 degrees C were similar, cells bearing temperature-sensitive mutations (mouse 135V or human 143A) were growth arrested at 31 degrees C. Temperature-dependent p53 activity was associated with increased levels of MDM2 and p21/WAF1, and the induction of an integrated p53-responsive luciferase gene. The REF lines exhibited similar sensitivities to common anticancer drugs when grown at 37 degrees C. However, when exposed to the same agents following transient incubation at 31 degrees C, the p53-arrested cells exhibited a marked survival advantage as shown by colony-forming assays. Chemoprotection was not universal, in that colony formation was not enhanced significantly after treatment with cisplatin or 5-fluorouracil, two drugs which can cause cellular damage throughout the cell cycle. Like other negative growth regulators, an activated p53 checkpoint may mediate the survival of cells exposed to drugs that target DNA synthesis or mitosis. PMID- 9351896 TI - Glutathione-dependent conversion to glyoxylate, a major pathway of dichloroacetate biotransformation in hepatic cytosol from humans and rats, is reduced in dichloroacetate-treated rats. AB - Although it has been postulated that glyoxylate is an intermediate in the biotransformation of DCA to oxalate, CO2, and glycine, there has been no positive identification of glyoxylate as a metabolite of DCA. We have demonstrated that a GSH-dependent pathway in dialyzed hepatic cytosol from rats and humans converts a mixture of 1-14C- and 1,2-13C-DCA to isotopically labeled glyoxylate. The reaction does not occur in the presence of NADPH or NADH or in the absence of GSH. The identity of the glyoxylate was demonstrated by HPLC with radiochemical detection and confirmed by GC/MS of the methylated glyoxylate, which showed the 13C-labeled product. The apparent Km for GSH was 0.075 mM in rat hepatic cytosol. Pretreatment of rats with NaDCA, 50 mg/kg, p.o. (by mouth) for 2 days prior to preparation of hepatic cytosol on the third day affected the cytosolic metabolism of DCA. With 0.2 mM DCA as substrate, in the presence of 1 mM GSH, control rats formed 1.45 +/- 0.13 nmol glyoxylate/min/mg cytosolic protein, whereas the rate in DCA-treated rats was 0.45 +/- 0.10 nmol glyoxylate/min/mg protein (mean +/- SD, N = 4 in each group). The mechanism of this reduction in the rate of DCA biotransformation in DCA-treated rats is unknown but is consistent with in vivo observations that the elimination of DCA from plasma of humans and rats is slowed by prior administration of DCA. PMID- 9351897 TI - Mechanisms of enhanced oral availability of CYP3A4 substrates by grapefruit constituents. Decreased enterocyte CYP3A4 concentration and mechanism-based inactivation by furanocoumarins. AB - Grapefruit juice increases the oral availability of a variety of CYP3A4 substrates. It has been shown that recurrent grapefruit juice ingestion results in a loss of CYP3A4 from the small bowel epithelium. We now show that the reduction in intestinal CYP3A4 concentration is rapid; a 47% decrease occurred in a healthy volunteer within 4 hr after consuming grapefruit juice. To identify the specific components of the juice responsible for this effect, we used a recently developed Caco-2 cell culture model of human intestinal epithelium that expresses catalytically active CYP3A4. We found that grapefruit oil and two furanocoumarin constituents (6', 7'-dihydroxybergamottin and a closely related dimer) caused a dose-dependent fall in CYP3A4 catalytic activity and immunoreactive CYP3A4 concentration. The effect was selective in that concentrations of CYP1A1 and CYP2D6 did not fall, consistent with previous results obtained in vivo. Assays of various juices confirmed that 6',7'-dihydroxybergamottin is the major furanocoumarin present and, although its concentration varies significantly among types and brands of grapefruit juice, it is consistently present in concentrations exceeding the IC50 (1 microM) for loss of midazolam 1'-hydroxylase activity determined in the Caco-2 cells. Studies with recombinant CYP3A4 revealed that 6', 7'-dihydroxybergamottin is a mechanism-based inactivator, which supports the idea that loss of CYP3A4 results from accelerated degradation of the enzyme. We conclude that the effect of grapefruit juice on oral availability of CYP3A4 substrates can be largely accounted for by the presence of 6',7' dihydroxybergamottin although other furanocoumarins probably also contribute. PMID- 9351898 TI - Heterologous expression of human drug-metabolizing enzymes. AB - This article is a report on a symposium held at the March 1997 meeting of the American Society for Pharmacology and Experimental Therapeutics in San Diego. Current developments in the heterologous expression of cytochrome P450, NADPH cytochrome P450 reductase, glutathione transferase, and UDP glucuronosyltransferase enzymes are described. Systems include bacteria, insect cells, and transient and stable mammalian cells. Uses of the products are described for discernment of which enzymes are involved in metabolism of drugs, genotoxicity assays, mutagenesis (for structure-activity relationships), large scale production of enzyme products, antibody production, and production of proteins for biophysical studies. PMID- 9351899 TI - Inactivation of cytochrome P450s 2B1, 2B4, 2B6, and 2B11 by arylalkynes. AB - The time-dependent loss of the 7-ethoxy-4-trifluoromethylcoumarin (EFC) O deethylase activity of rat P450 2B1, rabbit P450 2B4, or dog P450 2B11 by 1 ethynylnaphthalene (1EN), 2-ethynylnaphthalene (2EN), 2-(1-propynyl)naphthalene (2PN), 1-ethynylanthracene (1EA), 2-ethynylanthracene, 2-ethynylphenanthrene, 3 ethynylphenanthrene, 9-ethynylphenanthrene (9EPh), 9-(1-propynyl)phenanthrene (9PPh), 4-ethynylpyrene (4EP), and 4-(1-propynyl)biphenyl (4PbP) was investigated. The rate constants for inactivation by the arylalkynes in descending order of effectiveness for the top five compounds were 9EPh>9PPh>1EN, 2EN, 2PN for 2B1, 9EPh>2EN>4EP>1EN, 1EA for 2B4, and 9EPh>1EA>4EP, 9PPh>2EN for 2B11. The size and the shape of the aromatic ring system and the placement of the alkyne functional group were important for inactivation. The most effective inactivator with all the isozymes was 9EPh. This compound also inactivated the EFC activity in microsomes from human lymphoblastoid cells expressing human P450 2B6. The specificity of 9EPh for the inhibition or inactivation of different P450 activities in microsomes from rats treated with various inducing agents was determined by measuring lidocaine, testosterone, p-nitrophenol, or erythromycin metabolism. The greatest effect was observed with the 2B-specific products from lidocaine and testosterone, whereas no effect was seen with p-nitrophenol or erythromycin. When the covalent binding of [3H]2EN to microsomal protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, a radiolabeled protein band that corresponds to 2B1 was observed in the lanes containing microsomes from rats treated with phenobarbital and, to a lesser extent, pyridine and isosafrole after incubation with NADPH. When these microsomes were incubated with [3H]9EPh or [3H]1EP, two NADPH-dependent bands were radiolabeled. One corresponded to 2B1/2 and the other to a protein of approximately 59 kDa, which was observed in the lanes from phenobarbital-treated male and female rats and pyridine-treated male rats. No radiolabeled bands were observed with [3H,14C]4PbP with any of the microsomes. PMID- 9351900 TI - Gender, age and dose effects of neonatally administered aspartate on the sexually dimorphic plasma growth hormone profiles regulating expression of the rat sex dependent hepatic CYP isoforms. AB - Newborn male and female rat pups were injected with either 2 mg or 4 mg monosodium aspartate (MSA)/g body weight or diluent on alternate days for the first 9 days of life. Both doses of the amino acid had profound effects on the sexually dimorphic growth hormone secretory profiles in adulthood. There were no measurable levels of growth hormone in any of the plasma samples obtained during 8 continuous hr of serial blood collections from the adult males and females treated neonatally with 4 mg of MSA. Male rats treated with half the dose of the amino acid (i.e., 2 mg MSA/g) exhibited typical masculine profiles of growth hormone release, except that the amplitudes of the ultradian pulses were reduced to 10-20% of normal male levels. Otherwise, like normal males, the peaks occurred about every 3-4 hr and the intervening 2.5-hr troughs had undetectable levels of growth hormone. In a similar sense, females treated with 2 mg of MSA maintained their sexually dimorphic pattern of plasma growth hormone, i.e., frequent pulses of hormone followed by short-lived troughs. However, the peaks rarely exceeded 20 ng/ml and the troughs usually fell to a measurable 8 to 10 ng/ml resulting in an approximate 75% reduction in the mean plasma concentration. Growth hormone- and gender-dependent expression of CYP2C7, 2C11, 2C12, 2C13, 2A1, 2A2, and 3A2 (mRNAs, proteins, and catalytic activities) were generally unaffected by neonatal exposure to 2 mg of MSA. In contrast, the higher 4-mg dose of the amino acid completely or near completely suppressed male-specific CYP2C11, 2C13, 2A2, and 3A2 expression while inducing small increases in female-specific CYP2C12 and female-predominant CYP2A1 in the treated males. Females exposed to the 4 mg MSA dose exhibited less severe isoform changes characterized by small reductions in CYP2C12 and 2C7 levels. Whereas expression levels of most of the CYP isoforms in both sexes were lowest in the pubertal (47-day-old) rats, and occasionally higher in the adults (207-day-old) as compared with the early postpubertal (70-day-old) rats, the effects of neonatal MSA were the same at all ages studied. Since each of the CYP isoforms are regulated by different "signaling elements" in the sexually dimorphic plasma growth hormone profiles, it is possible to correlate MSA-induced alterations in CYP expression levels to specific changes in the gender-dependent growth hormone profiles. PMID- 9351901 TI - 14C-Propoxyphene demethylation in the rat. An example of differences between liver and intestinal drug-presystemic metabolism. AB - Presystemic metabolism is believed to occur mainly in the liver with some minor intestinal participation. The aim of this study was to investigate the respective part of each of these two organs in the metabolism of the analgesic d propoxyphene (DP). Pharmacological doses of DP were given in the duodenum (ID), the portal vein (IP), and the femoral vein (IV) of male Wistar rats. A tracer dose of 14C-DP was also administered either in IV, IP, or ID as well as in hepatectomized rats or rats with bile duct diversion. In vitro demethylation occurring in liver and intestinal microsomes was also studied. Absolute DP bioavailability obtained after oral administration was two times higher than that observed after portal administration (48.9% vs. 23.2%, respectively), an result opposite (i.e. a lower bioavailability) of that expected on the basis of the existence of a liver enzyme saturation phenomenon. The 14CO2 cumulative excretion after 14C-DP administration was significantly lower after IV or ID administration than after injection in the portal vein as a bolus or within 20 min. The biliary excretion of the labeled compound varied in the opposite direction, being greater after IV or ID than after IP administration, suggesting that the metabolism of DP in the liver is influenced by an extrahepatic transformation. This most likely occurs in the gut since the production of 14CO2 after IV administration was similar to that after ID administration. This transformation did not prohibit DP detection in the systemic blood but was sufficient to increase the part eliminated with bile and to decrease the part demethylated into NP. Demethylation mainly occurs in the liver since the production of 14CO2 was nearly abolished in hepatectomized rats. Furthermore, microsomes of hepatic but not of intestinal origin were able to demethylate DP. Our data suggest that the transformation of DP occurring in gut after oral administration is responsible for changes in the hepatic metabolism of the drug. PMID- 9351902 TI - Use of the deconvolution principle in the estimation of absorption and pre systemic intestinal elimination of drugs. AB - The deconvolution principle was used to evaluate the extent of absorption and first-pass elimination of selected drugs. In the first example, deconvolution of the portal blood profiles of etretinate (ET, a synthetic retinoid) indicated that there was significant gut-wall conversion of ET to acitretin (ETA, the primary metabolite of ET) during a 60-min intestinal perfusion of ET. In the second example, deconvolution was used to confirm that the extent of carbovir disappearing from the gastrointestinal lumen was matched by the extent of carbovir appearance in the portal blood. Thus, deconvolution has several important applications in the study of absorption and intestinal first-pass metabolism. PMID- 9351903 TI - Formation of guanoxabenz from guanabenz in human liver. A new metabolic marker for CYP1A2. AB - The in vitro N-hydroxylation of guanabenz as well as the corresponding N dehydroxylation of guanoxabenz has been previously detected in biotransformation studies with microsomal fractions of different species including human hepatic microsomes. Furthermore, the N-hydroxylation of guanabenz was found to be catalyzed by enriched cytochrome P450 (P450) fractions in reconstituted systems. Strong correlations between 7-ethoxyresorufin O-deethylation (r = 0. 96; p < 0.001), caffeine N-demethylation (r = 0.92; p < 0.001), respectively, and guanabenz N-hydroxylation activities were demonstrated in 10 human liver microsomal preparations. Studies with microsomes from human B-lymphoblastoid cell lines expressing human cytochrome P450 enzymes proved that CYP1A2 is the major isozyme responsible for this metabolic pathway. Further, P450 isozymes did not show any detectable conversion rates. The reaction was inhibited in presence of the potent CYP1A2 inhibitors alpha-naphthoflavone (7, 8-benzoflavone) and furafylline. The N-reduction of guanoxabenz to guanabenz exhibits a significant correlation to the benzamidoxime N-reduction after incubation with 10 human liver microsomal preparations (r = 0.97; p < 0.001). The formation of benzamidine from benzamidoxime was described previously to be catalyzed by the benzamidoxime reductase. These results suggest that the guanabenz N-hydroxylation is mediated via CYP1A2, whereas the corresponding guanoxabenz N-reduction is catalyzed by an enzyme system composed of cytochrome b5, NADH cytochrome b5-reductase, and benzamidoxime reductase. The high affinity of guanabenz to CYP1A2 and the distinct selectivity of this P450 isozyme toward guanabenz confirms the in vitro guanabenz N-hydroxylation to be a suitable metabolic marker for CYP1A2 in biotransformation studies. PMID- 9351904 TI - Pharmacokinetics and metabolism in mice of a phosphorothioate oligonucleotide antisense inhibitor of C-raf-1 kinase expression. AB - The plasma and tissue disposition of CGP 69846A (ISIS 5132) was characterized in male CD-1 mice following iv bolus injections administered every other day for 28 days (total of 15 doses). The doses ranged from 0.8 mg/kg to 100 mg/kg. Urinary excretion of oligonucleotide was also monitored over a 24-hr period following single dose administration over the same dose range. Pharmacokinetic plasma profiles were determined following single dose administration (dose 1) and after multiple doses (dose 15) at doses of 4 and 20 mg/kg. Concentrations in kidney, liver, spleen, heart, lung, and lymph nodes were characterized following doses 1, 8, and 15 for all doses. Capillary gel electrophoresis was used to quantitate intact (full-length) oligonucleotide and its metabolites (down to N - 11 base deletions) in both plasma and tissue at all time points. The plasma and tissue disposition of CGP 69846A was characterized by a rapid distribution into all tissues analyzed. Rapid plasma clearance of the parent oligonucleotide (9.3-14.3 ml/min/kg) was predominantly the result of distribution to tissue and, to a lesser extent, metabolism. Appearance and pattern of chain-shortened metabolites seen in plasma and tissue were consistent with predominantly exonuclease-mediated base deletion. No measurable accumulation of oligonucleotide was observed in plasma following multiple-dose administration, but both the liver and the kidney exhibited 2-3-fold accumulations. In general, the tissues exhibited half-lives for the elimination of parent oligonucleotide of 16-60 hr compared with plasma half-lives of 30-45 min. After repeated administrations, significant decreases in plasma clearance and volume of distribution at steady state (Vss) were observed following dose 15 at the dose of 20 mg/kg but not at the dose of 4 mg/kg. Changes in tissue accumulation and evidence for saturation of tissue distribution at the high doses may explain the plasma disposition changes observed in the absence of alteration of metabolism or plasma accumulation. Urinary excretion was a minor pathway for elimination of oligonucleotide over the 24-hr period immediately following iv administration. However, the amount of oligonucleotide excreted in the urine increased as a function of dose from less than 1% to approximately 13% of the administered dose over a dose range of 0.8 mg/kg to 100 mg/kg. PMID- 9351905 TI - Metabolic profiles of montelukast sodium (Singulair), a potent cysteinyl leukotriene1 receptor antagonist, in human plasma and bile. AB - Montelukast sodium [1-([(1(R)-(3-(2-(7-chloro-2-quinolinyl)-(E)- ethenyl)phenyl) 3-(2-(1-hydroxy-1-methylethyl)phenyl)propyl)thio]methyl)cyclopropylacetic acid sodium salt] (MK-476, Singulair) is a potent and selective antagonist of the cysteinyl leukotriene (Cys-LT1) receptor and is under investigation for the treatment of bronchial asthma. To assess the metabolism and excretion of montelukast, six healthy subjects received single oral doses of 102 mg of [14C]montelukast, and the urine and feces were collected. Most of the radioactivity was recovered in feces, with 90% of the total radioactivity present in urine, bile, and plasma. PMID- 9351907 TI - Metabolism of the antiandrogenic drug (Flutamide) by human CYP1A2. AB - The antiandrogenic drug, flutamide, is widely used in the treatment of carcinoma of the prostate. The present study examines the metabolism of flutamide by human liver microsomes and purified recombinant human cytochrome P450s (CYP), expressed as fusion proteins. These studies show the principal role of CYP1A2 in the metabolism of flutamide to 2-hydroxyflutamide. A minor metabolite is formed during the metabolism of flutamide by CYP3A4 in the presence of an excess of added purified NADPH-P450 reductase. The metabolism of flutamide is inhibited by low concentrations of alpha-naphthoflavone and ketoconazole. Other substrates of CYP1A2, such as phenacetin, imipramine, caffeine, and estradiol, are also inhibitors of flutamide metabolism by CYP1A2. Of interest is the inhibition of flutamide metabolism by its metabolite, 2-hydroxyflutamide, and the inhibition of the 2- and 4- hydroxylation of estradiol by flutamide. CV1 cells do not metabolize flutamide to 2-hydroxyflutamide. In assays performed using this cell line transfected with the cDNA for the androgen receptor, flutamide is a pure antagonist, and 2-hydroxyflutamide, while a more potent androgen receptor (AR) antagonist, activates the AR at higher concentrations. Stable expression of CYPIA2 in these CV1 cells causes flutamide to exhibit agonistic properties at higher concentrations, a behavior not exhibited by cells stably transfected only with the expression vector encoding the AR. These findings raise the possibility that increased conversion of flutamide to 2-hydroxyflutamide or accumulation of 2 hydroxyflutamide in cells may contribute to the anomalous responses to flutamide that are observed in some advanced prostate cancers. PMID- 9351908 TI - Studies on the interactions of chiral secondary alcohols with rat hydroxysteroid sulfotransferase STa. AB - Hydroxysteroid (alcohol) sulfotransferase STa catalyzes the 3'-phosphoadenosine 5'-phosphosulfate-dependent O-sulfonation of a diverse array of alcohols including neutral hydroxysteroids. Many of the secondary alcohols that interact with this sulfotransferase are the metabolic products of stereoselective oxidation or reduction reactions. The role that the stereochemistry of secondary alcohol substrates plays in the catalytic efficiency of STa was investigated with a series of chiral benzylic alcohols and the enantiomeric 3-hydroxyl-containing steroids, androsterone and epiandrosterone. In the case of (R)-(+)- and (S)-(-) enantiomers of 2-methyl-1-phenyl-1-propanol and 1-phenyl-1-butanol, the effect of stereochemistry on the catalytic efficiency of STa was small (less than 2-fold in favor of (R)-(+)-enantiomers). However, as the number of carbons in the alpha alkyl chain increased, the stereoselectivity for the sulfation of enantiomers increased as well. The (R)-(+)-enantiomers of 1-phenyl-1-pentanol, 1-phenyl-1 hexanol, and 1-phenyl-1-heptanol were preferred as substrates over the (S)-(-) enantiomers with a 3-fold difference in catalytic efficiency. STa showed absolute stereospecificity in the sulfation of the enantiomers of 1-phenyl-1 cyclohexylmethanol; (R)-(+)-1-phenyl-1-cyclohexylmethanol was a substrate for STa, while the (S)-(-)-enantiomer was a competitive inhibitor of the enzyme. Although a lower degree of stereoselectivity was observed with the 3-hydroxyl containing steroids, androsterone and epiandrosterone, results with these substrates were also consistent with the conclusion that the stereochemistry of secondary alcohols is an important factor in the catalytic efficiency of STa. PMID- 9351909 TI - Reversible formation of fatty acid esters of budesonide, an antiasthma glucocorticoid, in human lung and liver microsomes. AB - Microsomes from human lung and liver catalyze the formation of fatty acid esters of budesonide, a glucocorticoid used for inhalation treatment of asthma. The conjugation was dependent on coenzyme A and ATP. Addition of free fatty acids to the incubations affected the pattern of metabolites, but ester formation was observed also without such addition. Budesonide oleate, palmitate, linoleate, palmitoleate, and arachidonate were identified as metabolites. The fatty acid conjugates of budesonide were shown to be substrates for lipase in vitro, thus budesonide is regainable from the conjugates. The data suggest that an equilibrium between budesonide and these pharmacologically inactive lipoidal conjugates will be established in tissues at repeated exposure to budesonide. Since the fatty acid conjugates most likely will be retained intracellularly for a longer time than unchanged budesonide, the duration of tissue exposure to budesonide will depend partly on the rate of lipase-catalyzed hydrolysis of the conjugates. The findings in this study provide a possible explanation for the efficacy of budesonide in mild asthmatics also when inhaled once daily. PMID- 9351910 TI - Synthesis and reactivity of coumarin 3,4-epoxide. AB - Coumarin is used widely as a fragrance constituent and is administered clinically in the treatment of certain lymphedemas and malignancies. Although toxicity occurs only rarely in humans treated clinically with high-dose coumarin, it is well established that coumarin is hepatotoxic in the rat. This species difference in susceptibility to toxicity reflects the disparate metabolic processes occurring in humans and rodents. In humans, coumarin is converted extensively via cytochrome P450 2A6 to the nontoxic 7-hydroxycoumarin metabolite. In contrast, coumarin 3,4-epoxidation is thought to predominate in rodent species, resulting in the formation of several potentially toxic metabolites. Coumarin epoxide is thought to be highly unstable and has not been isolated synthetically or as a microsomal product. To address this issue, coumarin 3,4-epoxide was synthesized, and its stability and fate have been determined. Coumarin 3,4-epoxide was prepared by reacting coumarin with dimethyldioxirane. The epoxide was stable in organic solvents and survived conditions required for analysis by gas chromotography. Its structure was confirmed via 1H-NMR and gas chromatography mass spectrometry-infrared spectroscopy (GC-MS-IR). In contrast, coumarin 3,4 epoxide was unstable in aqueous solution, converting within 20 sec to a ring opened compound. Using GC-MS-IR analysis, the single coumarin 3,4-epoxide product was identified as o-hydroxyphenylacetaldehyde (o-HPA). Although other investigators have suggested that 3-hydroxycoumarin is an intermediate in o-HPA formation from coumarin 3,4-epoxide, we have demonstrated that 3-hydroxycoumarin, incubated in an aqueous system or with liver microsomal proteins, does not form o HPA. Thus, the results of the present work establish that coumarin 3,4-epoxide can be synthesized and that o-HPA, which has previously been shown to be a prominent coumarin metabolite in rat liver microsomal incubations, is formed directly from coumarin 3,4-epoxide. These results suggest that both coumarin 3,4 epoxide and o-HPA may contribute to the hepatotoxicity of coumarin. PMID- 9351911 TI - Pharmacokinetics of recombinant human insulin-like growth factor-I in diabetic rats. AB - Pharmacokinetics of recombinant human insulin-like growth factor-I (rhIGF-I) was investigated after iv administration (0.32, 1.0, and 3. 2 mg/kg) to normal and streptozotocin-induced diabetic rats. rhIGF-I was eliminated from plasma biexponentially in both normal and diabetic rats. Plasma concentrations of rhIGF I were lower at almost all the time points examined in diabetic rats than in normal rats. The pharmacokinetic parameters of total body clearance (CLtotal), mean residence time (MRT), and elimination rate constant (kel) indicated that rhIGF-I disappeared more rapidly in diabetic rats than in normal rats at any dosage. The amounts of IGF binding proteins (IGFBPs) in plasma were assessed by determining the endogenous IGF-I and. Levels of the 150 kDa complex, a ternary complex of IGF-I with IGFPB-3 and an acid-labile subunit, the 50 kDa complex, a complex of IGF-I with IGFBP-2, were found to be lower in diabetic rats than in normal rats. Fractions of rhIGF-I free and bound to the binding proteins were estimated by gel chromatographic separation of rhIGF-I in plasma after iv administration, and the pharmacokinetics of free and bound rhIGF-I was analyzed independently. Plasma concentrations of free and bound rhIGF-I were lower in diabetic rats than in normal rats, especially the concentrations of the 150 kDa complex were much lower. The reduced IGFBP-3 would be responsible for the faster elimination of rhIGF-I in diabetic rats. PMID- 9351965 TI - New light on TRP and TRPL. AB - Store-operated Ca2+ entry, a mode of Ca2+ influx activated by depletion of Ca2+ from the internal stores, has been detected in a wide variety of cell types and may be the primary mechanism for Ca2+ entry in nonexcitable cells. Nevertheless, until recently, no candidate store-operated channel (SOC) had been identified molecularly. Through the serendipity of Drosophila genetics, a candidate SOC, referred to as Transient Receptor Potential (TRP), has been identified that is essential for the light-induced cation conductance in photoreceptor cells. A combination of in vitro and in vivo studies has provided strong evidence that TRP is a bona fide SOC. Moreover, TRP forms a supramolecular complex, proposed to be critical for feedback regulation and/or activation, that includes rhodopsin, phospholipase C, protein kinase C, calmodulin, and the PDZ domain-containing protein, INAD. INAD seems to be a scaffolding protein that links TRP with several of these other proteins in the complex. TRP also complexes with a related channel subunit, TRP-like, to form a heteromultimer with conductance characteristics distinct from those of TRP or TRP-like homomultimers. A family of proteins related to TRP is conserved from Caenorhabditis elegans to humans, and recent evidence indicates that at least some of these proteins are SOCs. The human TRP related proteins may mediate many of the store-operated conductances that have been identified previously in a plethora of human cells. PMID- 9351966 TI - Subtype-specific differences in subcellular localization of alpha1-adrenoceptors: chlorethylclonidine preferentially alkylates the accessible cell surface alpha1 adrenoceptors irrespective of the subtype. AB - Selective inactivation of alpha1B-adrenoceptor (AR) by the site-directed alkylating agent chlorethylclonidine (CEC) has been used as one of major pharmacological criteria to subclassify alpha1-AR; however, the mechanism for the differential CEC sensitivity of the two subtypes is uncertain, and the extent of CEC inactivation varies depending on the treatment employed. In this study, we examined the correlation between the subcellular localization of alpha1-AR subtypes (alpha1A and alpha1B) and CEC sensitivity. Constructing alpha1-AR tagged with the FLAG epitope at the amino terminus and/or green fluorescent protein (GFP) at the carboxyl terminus, we examined the subcellular distribution of alpha1-ARs expressed in COS-7 cells. Flow cytometry analysis showed that most populations of GFP-expressing alpha1B-AR cells, but very few GFP-expressing alpha1A-AR cells, were detected by the anti-amino terminus antibodies. The immunocytochemical and GFP-fluorescence confocal micrographs showed that alpha1A ARs predominantly localize intracellularly, whereas alpha1B-ARs localize on the cell surface. Furthermore, CEC (10 microM) treatment of intact cells resulted in an inactivation of approximately 42% of alpha1A-ARs and 93% of alpha1B-ARs, whereas treatment of the membrane preparations resulted in an inactivation of approximately 83% of alpha1A-ARs and 88% of alpha1B-ARs, respectively. Together, the results showed that a hydrophilic alkylating agent CEC preferentially inactivates alpha1-AR on the cell surface irrespective of its subtype, and that the subtype-specific subcellular localization rather than the receptor structure is a major determinant for CEC inactivation of alpha1-AR. Subtype-specific subcellular localization suggests an additional class of functional properties that provide new insight into drug action. PMID- 9351967 TI - Mode of interaction of G-quartets with the integrase of human immunodeficiency virus type 1. AB - Oligonucleotides that can form a highly stable intramolecular four-stranded DNA structure containing two stacked guanosine-quartets (G-quartets) have been reported to inhibit the replication of the human immunodeficiency virus type 1 (HIV-1) in cell culture. Two possible mechanisms for the observed antiviral activity have been proposed: interference with virus adsorption to the cell and/or inhibition of HIV-1 integrase. We investigated the molecular interaction of G-quartet-containing oligonucleotides with HIV-1 integrase in comparison with random oligonucleotides and dextran sulfate. The prototypical G-quartet containing oligonucleotide, T30177 (Zintevir), inhibited the overall integration reaction with an IC50 value of 80 nM. A random oligonucleotide was 10-fold less potent, but dextran sulfate was more potent, with an IC50 value of 7 nM. We developed novel kinetic assays to dissect the overall integration reaction in three steps: the formation of the initial stable complex (ISC), the 3'-processing reaction, and the DNA strand-transfer step. We then analyzed the kinetics of the ISC formation and 3'-processing. The rate constant determined for the conversion of ISC into the cleaved product was 0.08 +/- 0.01 min-1. T30177 did not inhibit 3'-processing or DNA strand transfer, whereas dextran sulfate inhibited DNA strand transfer to some extent. Binding studies using surface plasmon resonance technology revealed that both T30177 and dextran sulfate were capable of preventing the binding of integrase to specific DNA. We propose a model in which the interaction of HIV-1 integrase with G-quartets results in the inhibition of the formation of the ISC between integrase and substrate DNA. Finally, we selected for an HIV-1 strain that was resistant to T30177 in cell culture. DNA sequence analysis revealed mutations in the envelope glycoprotein gp120 but not in the integrase gene. Although gp120 seems to be the main target for the antiviral activity in cell culture of G-quartets, the study of their specific inhibition of HIV-1 integrase may lead to the development of effective integrase inhibitors. PMID- 9351968 TI - The vascular smooth muscle type I angiotensin II receptor mRNA is destabilized by cyclic AMP-elevating agents. AB - Although processes involved in mRNA degradation play a significant role in dictating steady state mRNA levels, the influence of cell surface signaling on mRNA stability control is understood incompletely. In this study, the effects of cAMP-elevating agents on type I angiotensin II receptor (AT1-R) mRNA levels were assessed in cultured rat aortic vascular smooth muscle cells (VSMCs). AT1-R mRNA levels are rapidly reduced by forskolin treatment, in which the maximal effect yields an 80% reduction in AT1-R mRNA levels after 6 hr of treatment. The rate of AT1-R mRNA decay in response to forskolin is greater than its apparent intrinsic decay, as assessed in the presence of the transcriptional inhibitor 5,6-dichloro 1-beta-D-ribofuranosylbenzimidazole, suggesting forskolin treatment destabilizes the AT1-R mRNA. Nuclear run-on analysis indicates forskolin treatment does not affect transcription of the AT1-R gene in VSMCs, implying induced AT1-R mRNA destabilization accounts for the entire effect of forskolin in decreasing AT1-R mRNA levels. Dose-effect studies that assessed AT1-R mRNA levels and cAMP production were conducted using forskolin and the beta-adrenergic receptor agonist isoproterenol as agonists. Isoproterenol is almost 3 orders of magnitude more potent at eliciting the reduction in AT1-receptor mRNA levels than it is at stimulating cAMP production. Similarly, forskolin elicits reductions in AT1-R mRNA, which occur at concentrations that fail to elicit a detectable production of cAMP. However, protein kinase A activity is stimulated maximally by isoproterenol and forskolin concentrations that do not stimulate detectable cAMP production. These data provide evidence that the mechanism for down-regulation of AT1-R mRNA levels by cAMP-elevating agents in VSMCs occurs via a PKA-regulated mRNA destabilization pathway. PMID- 9351969 TI - Ligand-induced phosphorylation, clustering, and desensitization of A1 adenosine receptors. AB - Through immunocytochemistry with the use of antibodies against A1 adenosine receptors (A1Rs) and confocal microscopy, we show that stimulation of A1Rs by the agonist (R)-phenylisopropyladenosine [(R)-PIA] caused a rapid (5-15 min) aggregation (clustering) of receptor molecules on the surface of DDT1MF-2 cells. Internalization of the chronically stimulated receptor was slower and occurred concomitantly, with a time-dependent decrease (50%) in the number of cell surface [3H](R)-PIA binding sites. The reduction of binding sites was due partly (30%) to internalization and partly (20%) to the presence of desensitized cell surface receptor molecules that were unable to bind the ligand. Chronic exposure of DDT1MF-2 cells to 50 nM (R)-PIA produced functional desensitization, as deduced from second messenger production assays. Quantification of the content of A1Rs by immunoblotting and flow cytometry in cells pretreated with 50 nM (R)-PIA indicates a time-dependent slow down-regulation of the receptor. Receptor clustering and agonist-induced receptor phosphorylation, which occurred in serine and tyrosine, were simultaneous. The finding that activators of protein kinase A or C were able to induce functional desensitization of A1Rs, phosphorylate A1Rs in serine and threonine, and trigger clustering of the receptor suggests that phosphorylation of A1Rs in serine/threonine is involved in desensitization related events. PMID- 9351970 TI - Fludarabine-mediated repair inhibition of cisplatin-induced DNA lesions in human chronic myelogenous leukemia-blast crisis K562 cells: induction of synergistic cytotoxicity independent of reversal of apoptosis resistance. AB - We demonstrated previously that the nucleoside of fludarabine (F-ara-A), a clinically effective agent against chronic lymphocytic leukemia and low-grade lymphoma, produces synergistic cytotoxicity against cisplatin-resistant CP2.0 human colon tumor cells when administered in combination with cisplatin. The purpose of this study was 2-fold: (i) to determine whether the synergy occurs in K562 human chronic myelogenous leukemia cells, which, unlike CP2.0 cells, are relatively resistant to drug-induced apoptosis because they express P210(bcr-abl) and (ii) to study the underlying mechanism for the synergy if the enhancement of cytotoxicity occurs in K562 cells. When K562 cells were treated with fludarabine nucleoside and cisplatin as single agents for 4 hr, IC50 values for fludarabine and cisplatin were 3.33 and 2.28 microM, respectively, as measured by a clonogenic survival assay. The simultaneous treatment of K562 cells with the two agents resulted in synergistic cell killing as determined by median-effect analysis. Such synergistic cell killing by combined cisplatin and fludarabine could not be detected in repair-deficient human xeroderma pigmentosum cell lines. Within the range of cytotoxic concentrations, fludarabine (2.5-15 microM) and cisplatin (3-30 microM) as single agents produced no detectable internucleosomal DNA fragmentation as revealed by gel electrophoresis, nor did the combination of the two drugs induce apoptotic DNA degradation. The effects of fludarabine on the repair of cisplatin-induced DNA adducts and interstrand cross-links in K562 cells were analyzed to determine their correlation with the cytotoxic synergy. The interstrand cross-links were measured by the ethidium bromide binding fluorescence assay and quantitative Southern blotting technique. Repair of the intrastrand adducts was detected with whole-cell extracts using a cisplatin damaged plasmid as the substrate for the in vitro repair assay. Fludarabine at clinically achievable concentrations (1.5-4.5 microM fludarabine nucleoside; 20 100 microM fludarabine triphosphate) inhibited the repair of the DNA lesions induced by cisplatin in a dose-dependent fashion in K562 cells but not in xeroderma pigmentosum cells. Cotreatment with fludarabine preferentially increased the number of interstrand cross-links induced by cisplatin in actively transcribed genes in K562 cells. These data demonstrate the DNA-repair-inhibitory effect of fludarabine and suggest that this effect may contribute to the synergistic cytotoxicity of the fludarabine/cisplatin combination that resulted in decreased clonogenic survival of apoptosis-resistant K562 cells. PMID- 9351971 TI - Ligand binding pocket of the human somatostatin receptor 5: mutational analysis of the extracellular domains. AB - The ligand binding domain of G protein-coupled receptors for peptide ligands consists of a pocket formed by extracellular and transmembrane domain (TM) residues. In the case of somatostatin (SRIF), however, previous studies have suggested that the binding cavity of the octapeptide analog SMS201-995 (SMS) is lined by residues in TMs III-VII. The additional involvement of the extracellular domains for binding SMS or the natural SRIF ligands (SRIF-14, SRIF-28) has not been clarified. Using a cassette construct cDNA for the human somatostatin 5 receptor (sst5R), we systematically examined the role of exofacial structures in ligand binding by creating a series of mutants in which the extracellular portions have been altered by conservative segment exchange (CSE) mutagenesis for the extracellular loops (ECLs) and by deletion (for the NH2-terminal segment) or truncation analysis (ECL3). CHO-K1 cells were stably transfected with wild type or mutant human sst5R constructs, and agonist binding was assessed using membrane binding assays with 125I-LTT SRIF-28 ligand. Deletion of the NH2 terminus or CSE mutagenesis of ECL1 and ECL3 produced minor 2-8-fold decreases in affinity for SRIF-14, SRIF-28, and SMS ligands. Truncation of ECL3 to mimic the size of this loop in sst1R and sst4R (the two subtypes that do not bind SMS) did not interfere with the binding of SMS, SRIF-14, or SRIF-28. In contrast, both ECL2 mutants failed to bind 125I-LTT SRIF-28. Immunocytochemical analysis of nonpermeabilized cells with a human sst5R antibody revealed that the mutant receptors were targeted to the plasma membrane. Labeled SMS (125I-Tyr3 SMS) also failed to bind to the mutant ECL2 receptors. These results suggest a potential contribution of ECL2 (in addition to the previously identified residues in TMs III-VII) to the SRIF ligand binding pocket. PMID- 9351972 TI - Evidence for nucleotide excision repair as a modifying factor of O6-methylguanine DNA methyltransferase-mediated innate chloroethylnitrosourea resistance in human tumor cell lines. AB - We examined the O6-methylguanine-DNA methyltransferase (MGMT) protein as well as MGMT activity levels and the excision repair cross-complementing rodent repair deficiency gene, ERCC2 (XPD), protein levels in 14 human tumor cell lines not selected for chloroethylnitrosourea (CENU) resistance. These results were compared with 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU) cytotoxicity and UV light sensitivity. MGMT protein correlated significantly with MGMT activity (r = 0.9497, p = 0.0001). There was no significant linear correlation between BCNU cytotoxicity and MGMT content as determined by both Western analysis (r = 0.139, p = 0. 6348) and activity assay (r = 0.131, p = 0.6515). However, MGMT-rich cell lines were found to be more resistant than MGMT-poor cell lines to BCNU (t = 2.2375, p = 0.0225) but not to UV (t = 1.1734, p = 0.1317). Furthermore, the most BCNU-sensitive cell lines were all MGMT-poor. UV sensitivity was significantly correlated to BCNU cytotoxicity (r = 0.858, p = 0.0001). Significant correlations were found between ERCC2 protein levels and BCNU cytotoxicity (r = 0.786, p = 0.0009) or UV sensitivity (r = 0.874, p = 0.0001). Our results confirm that MGMT plays an important role in CENU resistance, but not in UV resistance. The correlation of UV sensitivity with BCNU cytotoxicity suggests that nucleotide excision repair is an important modifying factor of MGMT-mediated innate CENU resistance in human tumor cell lines, especially in highly resistant cell lines. ERCC2 may be implicated in this process. PMID- 9351973 TI - Phosphorylation of the Kv2.1 K+ channel alters voltage-dependent activation. AB - The voltage-gated delayed-rectifier-type K+ channel Kv2.1 is expressed in high density clusters on the soma and proximal dendrites of mammalian central neurons; thus, dynamic regulation of Kv2.1 would be predicted to have an impact on dendritic excitability. Rat brain Kv2.1 polypeptides are phosphorylated extensively, leading to a dramatically increased molecular mass on sodium dodecyl sulfate gels. Phosphoamino acid analysis of Kv2.1 expressed in transfected cells and labeled in vivo with 32P shows that phosphorylation was restricted to serine residues and that a truncation mutant, DeltaC318, which lacks the last 318 amino acids in the cytoplasmic carboxyl terminus, was phosphorylated to a much lesser degree than was wild-type Kv2.1. Whole-cell patch-clamp studies showed that the voltage-dependence of activation of DeltaC318 was shifted to more negative membrane potentials than Kv2.1 without differences in macroscopic kinetics; however, the differences in the voltage-dependence of activation between Kv2.1 and DeltaC318 were eliminated by in vivo intracellular application of alkaline phosphatase, suggesting that these differences were due to differential phosphorylation. Similar analyses of other truncation and point mutants indicated that the phosphorylation sites responsible for the observed differences in voltage-dependent activation lie between amino acids 667 and 853 near the distal end of the Kv2.1 carboxyl terminus. Together, these parallel biochemical and electrophysiological results provide direct evidence that the voltage-dependent activation of the delayed-rectifier K+ channel Kv2. 1 can be modulated by direct phosphorylation of the channel protein; such modulation of Kv2.1 could dynamically regulate dendritic excitability. PMID- 9351975 TI - Mitindomide is a catalytic inhibitor of DNA topoisomerase II that acts at the bisdioxopiperazine binding site. AB - The antitumor drug mitindomide (NSC 284356) was shown to inhibit the decatenation activity of human and Chinese hamster ovary (CHO) topoisomerase II [DNA topoisomerase (ATP-hydrolyzing), EC 5.99.1.1]. Mitindomide did not induce the formation of topoisomerase II-DNA covalent cleavable complexes in CHO cells. These results taken together indicate that mitindomide is a catalytic/noncleavable complex-forming-type inhibitor of topoisomerase II. The growth inhibitory effects of mitindomide and dexrazoxane toward a sensitive parent CHO cell line and the dexrazoxane-resistant DZR cell line, which is highly (500-fold) resistant to the bisdioxopiperazine dexrazoxane, were measured. The DZR cell line was shown to be 30-fold cross-resistant to mitindomide. Mitindomide, like dexrazoxane, was shown to inhibit cleavable complex formation by the topoisomerase II poison etoposide. The attenuated inhibition of etoposide induced cleavable complexes in DZR compared with CHO cells was, likewise, very similar for dexrazoxane and mitindomide. Together these results suggest that mitindomide acts at the same site on topoisomerase II as does dexrazoxane and other bisdioxopiperazines. Various molecular parameters obtained by molecular modeling were compared for mitindomide and dexrazoxane. Mitindomide, which is conformationally very rigid, has highly coplanar imide rings, as does dexrazoxane in the solid state. Other molecular parameters, such as the imide nitrogen-to imide nitrogen bond distances, and polar and nonpolar surface areas were also very similar. Thus, it is concluded that mitindomide exerts its antitumor effects through its inhibition of topoisomerase II by binding to the bisdioxopiperazine binding site. PMID- 9351974 TI - The interaction of arginine 106 of human prostaglandin G/H synthase-2 with inhibitors is not a universal component of inhibition mediated by nonsteroidal anti-inflammatory drugs. AB - The three-dimensional cocrystal structures of ovine prostaglandin G/H synthase-1 (PGHS-1) with S-flurbiprofen and murine PGHS-2 with S-flurbiprofen and indomethacin reveal that the carboxylate acid groups of these nonsteroidal anti inflammatory drugs (NSAIDs) form a salt bridge with the guanidinium group of Arg120 in PGHS-1 and Arg106 in PGHS-2. Mutagenesis studies confirmed that the Arg120 residue of PGHS-1 is critical for binding of substrate and inhibitors through ionic interactions of its guanidinium group with the carboxylate moieties of arachidonic acid and certain NSAIDs. We report here that the analogous R106E substitution in human PGHS-2 results in a catalytically active enzyme with a 30 fold higher Km value for arachidonic acid. Comparison of the inhibition of hPGHS 2(R106E) with wild-type hPGHS-2 by 11 structurally diverse selective and nonselective PGHS inhibitors revealed a 0-1000-fold decrease in inhibitory potency on the mutant enzyme. The loss of inhibitory potency of NSAIDs on hPGHS 2(R106E) could not be correlated with the presence or absence of a carboxylate functional group in the inhibitor, as was demonstrated previously for the PGHS 1(R120E) mutant, or with the selective or nonselective nature of the PGHS inhibitor. The decreases in the inhibitory potencies on hPGHS-2(R106E) by the carboxylate-containing NSAIDs flurbiprofen, indomethacin, meclofenamic acid, and diclofenac on hPGHS-2(R106E) were 965-, 48-, 5.5-, and 4.5-fold, respectively. The nonuniversal requirement for interaction of the carboxylate group of certain NSAIDs with the Arg106 residue in hPGHS-2 is supported by the observation that the methyl ester derivative of indomethacin was a more potent inhibitor than indomethacin on both hPGHS-2 and hPGHS-2(R106E). The greatest loss of potency for inhibition of hPGHS-2(R106E) was observed with the hPGHS-2-selective sulfonamide containing inhibitors NS-398 and flosulide. The PGHS-2-selective inhibitor DuP697 and a desbromo-sulfonamide analogue of DuP697 displayed equivalent potency on hPGHS-2(R106E) and hPGHS-2. The change in inhibitory potency of NS-398 on hPGHS 2(R106E) was due to a difference in the kinetics of inhibition, with NS-398 displaying time-dependent inhibition of hPGHS-2 but time-independent inhibition of PGHS-2(R106E). The time-dependent inhibition of hPGHS-2 by DuP697 was not affected by the presence of the R106E mutation. We conclude that the Arg106 residue of hPGHS-2 is involved in binding arachidonic acid and certain NSAIDs, but interactions with Arg106 are not a universal requirement for inhibition by either carboxylate-containing NSAIDs or PGHS-2-selective inhibitors. PMID- 9351976 TI - Canine mast cell adenosine receptors: cloning and expression of the A3 receptor and evidence that degranulation is mediated by the A2B receptor. AB - We cloned and characterized the canine A3 adenosine receptor (AR) and examined AR induced degranulation of the BR line of canine mastocytoma cells. Canine A3AR transcript is found predominantly in spleen, lung, liver, and testes and encodes a 314-amino acid heptahelical receptor. 125I-N6-Aminobenzyladenosine binds to two affinity states of canine A3AR with KD values of 0.7 +/- 0.1 and 16 +/- 0.8 nM, reflecting G protein-coupled and -uncoupled receptors, respectively. Xanthine antagonists bind with similar affinities to human, canine, and rabbit receptors but with 80-400-fold lower affinities to rat A3AR. Although canine BR mastocytoma cells contain A1AR, A2BAR, and A3AR, degranulation seems to be mediated primarily by A2BARs stimulated by the nonselective agonist 5'-N-ethylcarboxamidoadenosine (NECA) but not by the A3-selective agonist N6-(3-iodobenzyl)adenosine-5'-N methylcarboxamide. NECA-stimulated degranulation is not prevented by pertussis toxin and is blocked by enprofylline (Ki = 7 microM), an antiasthmatic xanthine with low affinity (Ki > 100 microM) for A1AR, A2AAR, and A3AR. NECA increases canine mastocytoma cell cAMP, Ca2+, and inositol trisphosphate levels; these responses are antagonized half-maximally by 7-15 microM enprofylline. The results suggest that (i) the cloned canine A3AR is structurally and pharmacologically more similar to human than to rat A3AR; (ii) the A2BAR, and not the A1AR or A3AR, is principally responsible for adenosine-mediated degranulation of canine BR mastocytoma cells; and (iii) the BR cell A2BAR couples to both Ca2+ mobilization and cAMP accumulation. Although A2B receptors play a major role in the regulation of BR mast cell degranulation, multiple AR subtypes and G proteins may influence mast cell functions. PMID- 9351977 TI - N-Glycosylation is not a prerequisite for glutamate receptor function but Is essential for lectin modulation. AB - All ionotropic glutamate receptor (iGluR) subunits analyzed so far are heavily N glycosylated at multiple sites on their amino-terminal extracellular domains. Although the exact functional significance of this glycosylation remains to be determined, it has been suggested that N-glycosylation may be a precondition for the formation of functional ion channels. In particular, it has been argued that N-glycosylation is required for the formation of functional ligand binding sites. We analyzed heterologously expressed recombinant glutamate receptors (GluRs) of all three pharmacological subclasses of glutamate receptors, N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, and kainate receptors. By expressing the GluR subunits in tunicamycin-treated, nonglycosylating Xenopus laevis oocytes, we determined that in neither case is N glycosylation required for ion channel function, although for NMDA receptors, functional expression in the absence of N-glycosylation is very low. Furthermore, we analyzed and compared the interaction of the desensitization-inhibiting lectin concanavalin A (ConA) with all functional GluR subunits. We show that although ConA has its most pronounced effects on kainate receptors, it potentiates currents at most other receptor subtypes as well, including certain NMDA receptor subunits, although to a much lesser extent. One notable exception is the alpha amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor GluR2, which is not affected by ConA. Furthermore, we show that ConA acts directly via binding to the carbohydrate side chains of the receptor protein. PMID- 9351978 TI - Key amino acids in the gamma subunit of the gamma-aminobutyric acidA receptor that determine ligand binding and modulation at the benzodiazepine site. AB - Pharmacological analyses of gamma-aminobutyric acidA (GABAA) receptor subtypes have suggested that both the alpha and gamma subunits, but not the beta subunit, contribute to the benzodiazepine binding site. We took advantage of the different pharmacological properties conferred by the inclusion of different gamma subunits in the receptor macromolecule to identify amino acids gamma2Phe77 and gamma2Met130 as key determinants of the benzodiazepine binding site. gamma2Phe77 was required for high affinity binding of the benzodiazepine site ligands flumazenil, CL218,872, and methyl-beta-carboline-3-carboxylate but not flunitrazepam. This amino acid was, however, required for allosteric modulation by flunitrazepam, as well as other benzodiazepine site ligands. In contrast, gamma2Met130 was required for high affinity binding of flunitrazepam, clonazepam, and triazolam but not flumazenil, CL218, 872, or methyl-beta-carboline-3 carboxylate and did not affect benzodiazepine efficacy. Introduction of the phenylalanine and methionine into the appropriate positions of gamma1 was not sufficient to confer high affinity for the benzodiazepine site ligand zolpidem. These data show that gamma2Phe77 and gamma2Met130 are necessary for high affinity binding of a number of benzodiazepine site ligands. Although most previous studies have focused on the contribution of the alpha subunit, we demonstrated a critical role for the gamma subunit at the benzodiazepine binding site, indicating that this modulatory site is located at the interface of these two subunits. Furthermore, gamma2Phe77 is homologous to alpha1Phe64, which has been previously shown to be a key determinant of the GABA binding site, suggesting a conservation of motifs between different ligand binding sites on the GABAA receptor. PMID- 9351979 TI - Epirubicin-induced oxidative DNA damage and evidence for its repair in lymphocytes of cancer patients who are undergoing chemotherapy. AB - Anthracycline derivatives have been widely used in the treatment of several types of human malignancies. Cytotoxicity of these drugs has been attributed to inhibition of topoisomerase II as well as intracellular production of free radicals. In our work we used a gas chromatography/mass spectrometry technique to study free radical-induced DNA base modifications in chromatin isolated from lymphocytes of cancer patients who received chemotherapy with epirubicin (one of anthracycline's antitumor derivatives). The anticancer therapy caused significant increases in the amount of all four DNA base modifications over control levels in the lymphocytes of most of the patients. For the majority of the cases the base products returned to the control value 24 hr after the infusion of the drug, which suggests the removal of these lesions by cellular repair processes. However, some of the modified bases escaped repair. Because part of these modifications may possess premutagenic properties, they may be responsible for secondary cancers induced by chemotherapy. PMID- 9351980 TI - Potentiation and inhibition of neuronal nicotinic receptors by atropine: competitive and noncompetitive effects. AB - Atropine, the classic muscarinic receptor antagonist, inhibits ion currents mediated by neuronal nicotinic acetylcholine receptors expressed in Xenopus laevis oocytes. At the holding potential of -80 mV, 1 microM atropine inhibits 1 mM acetylcholine-induced inward currents mediated by rat alpha2beta2, alpha2beta4, alpha3beta2, alpha3beta4, alpha4beta2, alpha4beta4, and alpha7 nicotinic receptors by 12-56%. Inward currents induced with a low agonist concentration are equally inhibited (alpha3beta2, alpha3beta4), less inhibited (alpha2beta4, alpha7), or potentiated (alpha4beta2, alpha4beta4) by 1 microM atropine. Effects on the more sensitive alpha4beta4 nicotinic receptors were investigated in detail by systematic variation of acetylcholine and atropine concentrations and of membrane potential. At high agonist concentration, atropine inhibits alpha4beta4 nicotinic receptor-mediated ion current in a noncompetitive, voltage-dependent way with IC50 values of 655 nM at -80 mV and of 4.5 microM at 40 mV. At low agonist concentration, 1 microM atropine potentiates alpha4beta4 nicotinic receptor-mediated ion current. This potentiating effect is surmounted by high concentrations of acetylcholine, indicating a competitive interaction of atropine with the nicotinic receptor, and potentiation is also reversed at high atropine concentrations. Steady state effects of acetylcholine and atropine are accounted for by a model for combined receptor occupation and channel block, in which atropine acts on two distinct sites. The first site is associated with noncompetitive ion channel block. The second site is associated with competitive potentiation, which appears to occur when the agonist recognition sites of the receptor are occupied by acetylcholine and atropine. The apparent affinity of atropine for the agonist recognition sites of the alpha4beta4 nicotinic acetylcholine receptor is estimated to be 29.9 microM. PMID- 9351981 TI - Down-regulation of mu-opioid receptor by full but not partial agonists is independent of G protein coupling. AB - In C6 glial cells stably expressing rat mu-opioid receptor, opioid agonist activation is negatively coupled to adenylyl cyclase through pertussis toxin sensitive G proteins. In membranes, [D-Ala2, N-MePhe4,Gly-ol5]enkephalin (DAMGO) increases guanosine-5'-O-(3-[35S]thio)triphosphate (GTP[gamma-35S]) binding by 367% with an EC50 value of 28 nM. Prolonged exposure to agonists induced desensitization of the receptor as estimated by a reduction in the maximal stimulation of GTP[gamma-35S] binding by DAMGO and rightward shifts in the dose response curves. In cells treated with 10 microM concentrations of etorphine, DAMGO, beta-endorphin, morphine, and butorphanol, DAMGO-stimulated GTP[gamma-35S] binding was 58%, 149%, 205%, 286%, and 325%, respectively. Guanine nucleotide regulation of agonist binding was correspondingly lower in membranes from tolerant cells. Furthermore, chronic opioid treatment increased forskolin stimulated adenylyl cyclase activity, and potency of DAMGO to inhibit cAMP accumulation was lower in morphine- and DAMGO-tolerant cells (EC50 = 55 and 170 nM versus 18 nM for control). Chronic treatment with agonists reduced [3H]DAMGO binding in membranes with the rank order of etorphine > DAMGO = beta-endorphin > morphine > butorphanol, and the affinity of DAMGO in alkaloid- but not peptide treated membranes was significantly lower in comparison with control. Pertussis toxin treatment of the cells before agonist treatment did not prevent the down regulation by full agonists; DAMGO and etorphine exhibited approximately 80% internalization, whereas the ability of partial agonists was greatly impaired. In addition to establishing this cell line as a good model for further studies on the mechanisms of opioid tolerance, these results indicate important differences in the inactivation pathways of receptor triggered by full and partial agonists. PMID- 9351982 TI - Expression of methylthioadenosine phosphorylase cDNA in p16-, MTAP- malignant cells: restoration of methylthioadenosine phosphorylase-dependent salvage pathways and alterations of sensitivity to inhibitors of purine de novo synthesis. AB - 5'-Deoxy-5'-methylthioadenosine phosphorylase (MTAP) is involved in the salvage of adenine and methylthio moieties of 5'-deoxy-5'-methylthioadenosine, a byproduct of polyamine synthesis, to adenine nucleotides and methionine, respectively. The gene encoding MTAP, MTAP, is frequently codeleted along with the tumor suppressor gene p16 in malignant cells bearing homozygous deletions in the chromosome 9p21 region. p16-, MTAP- malignant cells have been shown to be more susceptible to the purine de novo inhibitory actions of antifolates such as methotrexate than are p16+, MTAP+ cells. To understand the underlying mechanism, we reintroduced MTAP activity into two p16-, MTAP- cell model systems, the MiaPaCa-2 and PANC-1 human pancreatic carcinoma cell lines, by transfection with MTAP cDNA. It was found that transfection with MTAP cDNA (i) restored both the MTAP-dependent adenine and methionine salvage pathways, (ii) decreased the rates of purine de novo synthesis (18-47% lower than the wild-type or sham-transfected counterparts), and (iii) decreased cellular sensitivity to the antipurine-related growth-inhibitory actions of methotrexate and azaserine. These data support the hypothesis that operation of the MTAP-dependent adenine salvage pathway renders MTAP+ cells less dependent on de novo purine synthesis and hence less susceptible than MTAP- malignant cells to the growth-inhibitory actions of agents (e.g. antifolates) whose mechanism of action in part involves the de novo purine pathway. These findings provide a theoretical basis for the relatively selective action certain antifolates may have against MTAP-deficient malignancies. PMID- 9351983 TI - Role of free radicals in primary nonfunction of marginal fatty grafts from rats treated acutely with ethanol. AB - Acute treatment with one large dose of ethanol, which mimics binge drinking, causes marginal fatty liver and decreases survival significantly after liver transplantation in rats, yet mechanisms remain unclear. Therefore, we evaluated the possible role of free radicals in primary nonfunction caused by acute ethanol. Female donor rats were administered ethanol (5 g/kg orally) 20 hr before explantation, and grafts were stored in UW cold storage solution for 24-42 hr before implantation. Free radicals were trapped with alpha-(4-pyridyl 1-oxide)-N tert-butylnitrone after transplantation, and adducts were detected using electron spin resonance spectrometry. Ethanol increased a carbon-centered radical adduct in bile approximately 2-fold and elevated serum lipid hydroperoxides approximately 4-fold. Ethanol also increased transaminase release 3.7-fold and decreased bile production by 55%. Catechin, a free radical scavenger, minimized the increase in free radicals, blunted transaminase release, and elevated bile production significantly, indicating that free radical production plays an important role in ethanol-induced fatty graft injury. GdCl3 (20 mg/kg intravenously), a selective Kupffer cell toxicant, largely blocked the increases in free radical and lipid hydroperoxide production caused by ethanol. In addition, ethanol nearly doubled white blood cell adhesion after transplantation, leading to increased superoxide production in fatty grafts. GdCl3 largely blocked leukocyte adhesion as well as superoxide production. Allopurinol, an inhibitor of xanthine oxidase, also diminished free radical production, blunted transaminase release, and improved bile production in fatty grafts significantly. Taken together, we conclude that free radical formation increases in ethanol-induced fatty grafts due mainly to activation of Kupffer cells and increased adhesion of white blood cells. Antioxidants can effectively block free radical formation and minimize injury to marginal fatty grafts caused by binge drinking. PMID- 9351984 TI - The European Brain Injury Consortium. Nemo solus satis sapit: nobody knows enough alone. PMID- 9351985 TI - Evaluation of hemodynamic responses in head injury patients with transcranial Doppler monitoring. AB - Transcranial Doppler (TCD) can monitor middle cerebral artery (MCA) velocity which can be recorded simultaneously with other physiologic parameters such as end tidal (Et) CO2, arterial blood pressure and intracranial pressure (ICP), in head injured patients. Relative changes in MCA velocity can be used to reflect relative MCA blood flow changes during ICP waves, and also to evaluate cerebral autoregulation, CO2 reactivity and hemodynamic responses to mannitol and barbiturates. The utility and practicality of short intervals of TCD monitoring to evaluate hemodynamic responses, was evaluated in a group of 22 head injured patients (average Glasgow coma score 6). During ICP A waves, MCA velocity always decreased during the peak of the wave, and during ICP B waves, fluctuated synchronously with the ICP. Dynamic cerebral autoregulation, and reactivity to CO2, were reduced within 48 hours of admission. Impaired cerebral autoregulation within 48 hours of admission did not correlate with outcome at 1 month. Mannitol infusion caused an increase in MCA velocity (15.4 +/- 7.9%) which was significantly correlated to the impairment of dynamic autoregulation (r = 0.54, p < 0.0001). The MCA velocity response to a test dose of barbiturates was significantly correlated to the ICP (r = 0.61, p < 0.01) response as well as to the CO2 reactivity (r = 0.37, p < 0.05). Continuous MCA velocity monitoring using TCD may be useful in evaluating a variety of hemodynamic responses in head injury patients and may replace more cumbersome cerebral blood flow techniques which have been used in the past for these purposes. PMID- 9351986 TI - Chronic subdural haematoma--a comparison of two different treatment modalities. AB - Burr-hole craniotomy (BHC) and closed-system drainage undoubtedly is currently the most accepted treatment offered in chronic subdural haematoma (CSDH). Although twist-drill trephination (TDT) techniques have been available for years, now a special subdural catheter kit has been launched for treatment of CSDH. In a prospective study, 33 patients with 36 CSDH were treated with a 5-mm TDT regimen and insertion of a CORDIS subdural catheter (CORDIS Corp., Miami, USA). The results are compared with a consecutive series of 33 patients treated previously with an 11-mm BHC and closed-system drainage for 40 CSDH: Recurrence and persistence rate of CSDH treated with TDT necessitating a second intervention was 18.1%, no further surgical intervention was necessary. In BHC treated patients, 33.3% of haematomas had to be reoperated on, another 6.0% had to be re-operated on a third time. Infection rate in BHC treated patients was 18.1% as compared with a 0% infection rate in patients treated with the TDT technique. Mortality rate for the BHC method was 9.0% as compared with 6.0% in the TDT treatment regimen. Significantly better clinical results are achieved using the TDT technique with insertion of a special subdural catheter, making this procedure superior to the BHC regimen. PMID- 9351987 TI - Detailed evaluation of 2959 allogeneic and xenogeneic dense connective tissue grafts (fascia lata, pericardium, and dura mater) used in the course of 20 years for duraplasty in neurosurgery. AB - Surgical experience with 2959 allogeneic and xenogeneic dense connective tissue grafts (1767 of fascia lata, 909 of pericardium, and 283 of dura mater), used in 2665 neurosurgical operations performed in the course of 20 years (1976 to 1995) is reported. Duraplasty using either allogeneic or xenogeneic grafts has had a similar, and favourable clinical outcome. Nevertheless, the pliable deep frozen fascia lata grafts, which could be used in any location, have been reserved for sella turcica plugging, anterior cranial base plasty, aneurysmal wrapping, and surgery of lipomyelomeningocele. Pericardium and dura mater grafts were in the majority of cases used over the brain convexity and posterior cranial fossa. Ovine pericardium proved to be superior to bovine and allogeneic pericardia because of its workability, flexibility, reduced thickness, and better transparency. Postsurgical complications occurred in 7.3%, and they were: 1) cerebrospinal fluid fistulas in 2.8%; 2) meningites in 2.3% (aseptic 1.4%, bacterial 0.8%, and tumoural 0.1% meningites); 3) pseudomeningoceles in 2.2%; 4) wound infections in 0.6%; 5) malresorptive hydrocephalus in 0.5%; and 6) adhesions to nerve tissue in 0.5%. The majority of complications healed without surgery. Forty-eight grafts (1.6%) failed to fulfil the requirements of the surgeon, and 46 of them were re-operated upon. Though another thirty-nine grafts healed successfully, 39 shunts (1.5%) had to be performed for malresorptive hydrocephalus (0.9%), and/or for a big pseudomeningocele (0.6%). So, the pure complication rate in 2665 duraplasties was 3.1%. The complex evaluation of the allogeneic and xenogeneic grafts (fascia, pericardium, and dura mater), used for duraplasty in neurosurgery during the last 20 years proved them, as remarkably good, with a success rates of 96.9%. PMID- 9351988 TI - Olfactory neuroblastoma: detection of genomic imbalances by comparative genomic hybridization. AB - Olfactory neuroblastoma (esthesioneuroblastoma) is a very rare tumour of the olfactory mucosa. Morphological features and cytogenetic studies strongly suggest a neuro-ectodermal origin. Up to now, cytogenetic studies are inconsistent. Some of them have proposed that the tumour belongs to the pPNET family. In the present study we describe genomic imbalances in olfactory neuroblastoma in a 46-year-old woman by using the molecular cytogenetic technique--comparative genomic hybridization (CGH)--in order to define the spectrum of genetic abnormalities in the tumour. The anatomical location and morphological findings were the basis for the diagnosis of esthesionearoblastoma. Immunohistochemical reactions for NSE, synaptophysin, chromogranin A, HNK-1/Leu-7 and S-100 revealed a characteristic immunophenotype. The CGH analysis showed multiple changes including DNA overrepresentations of chromosomes 4, 8, 11 and 14, partial DNA gains of the long arms of chromosomes 1 and 17, deletions of the entire chromosomes 16, 18, 19 and X, and partial losses of chromosomes 5q and 17p. This study represents an early utilisation of the CGH technique in olfactory neuroblastoma and demonstrates that the tumour carries complex chromosomal aberrations. PMID- 9351989 TI - Apoptosis in astrocytic neoplasms. AB - Apoptosis is a form of programmed cell death characterized by specific morphologic and biochemical properties. Tumorgenesis is the consequence not only of cell proliferation but also the loss of the ability to undergo apoptosis [2]. Bcl-2 is a protooncogene which has the ability to block apoptosis in many cell types. Astrocytic neoplasms are very aggressive tumors which many times fail to respond to surgery, radiation or chemotherapy. They frequently overexpress wild type p53 which is associated with the expression of bcl-2, and thus they may have evolved a mechanism to subvert apoptosis and allow continued growth. We examined the apoptotic index in fifty-nine astrocytic tumors of various histological grades (Oncor ApopTag Plus In Situ Detection Kit) and compared this with the level of bcl-2 expression. Low grade astrocytomas (0.21 +/- 0.05; range 0.0-0.9) and anaplastic astrocytomas (0.27 +/- 0.13; range 0.0-2.6) had significantly less apoptosis than glioblastomas (0.70 +/- 0.13; range 0.0-2.1; Kruskal-Wallis test, P < or = 0.01). In contrast, bcl-2 expression was similar in all grades of astrocytic tumors and did not correlate with the apoptotic index. Cells of low grade and anaplastic astrocytomas are less likely to undergo apoptosis; however, this does not seem to be a direct consequence of the regulation of bcl-2 expression. The difference in growth potential despite differences in apoptotic index is likely to be attributed to differences in mitotic not apoptotic activity. PMID- 9351990 TI - Absence of apoptosis in somatotropinomas treated with octreotide. AB - Octreotide is a potent agonist of somatostatin that lowers the serum level of growth hormone (GH), and reduces the size of somatotropinomas. However, the detailed mechanism of shrinkage of this tumour is not known. We, therefore, evaluated 11 patients with somatotropinomas who were treated with octreotide 300 micrograms/day for 2-5 weeks to observe the morphological changes in the tumour using electron microscopy and the immunocytochemical study of apoptosis using polyclonal anti-single stranded DNA. Findings were compared with those obtained with bromocriptine treatment (10 mg/day, 2 weeks) of 5 patients with somatotropinomas, and 11 patients who received no preoperative treatment (control group). The octreotide group showed neither increase in stromal tissue nor cell death. The size of tumour cells appeared to be slightly reduced. No typical apoptotic bodies were seen on the electron micrographs. The apoptotic index in the octreotide group (0.40 +/- 0.60%; mean +/- SD) resembled that in the control group (0.81 +/- 0.79%). In contrast, the bromocriptine group showed some cell death and an increase in stromal tissue. The bromocriptine group also showed the apoptotic index which (20.1 +/- 14.8%) was significantly higher than that of the control group (0.81 +/- 0.79%). Thus, octreotide did not induce apoptosis in somatotropinomas despite the presence of tumour shrinkage. Because of the lack of fibrosis observed in the octreotide-treated tumours, the preoperative administration of octreotide may help to improve the outcome of the transsphenoidal operation. PMID- 9351991 TI - Surgical treatment of anterior skull base tumours. AB - Skull base tumours represent a special challenge to surgeons due to the complex anatomy of the area. While small tumours are easy to remove, large lesions can pose complex situations. The most difficult aspects are not only the approach and removal, but specially the repair of the defects created by the resection of the tumour. We present here our experience with the surgical removal of tumours on the anterior skull base. To achieve a good approach, we resort to a bifrontal craniotomy including the cilliar arches. To obtain a skull base bone flap that can be used for repair at the end of the procedure, we remove the roof of the nose and a part of the medial wall and roof of both orbits. While the tumour is removed, the skull base bone flap is autoclaved to kill all tumoural cells. At the end of the procedure this bone flap is replaced, wrapped with a flap of pericranium. Provided no orbit needs to be emptied, no other flap is needed to reconstruct the area. One advantage is that the surgical cavity is not occluded with tissues, thus facilitating early identification of any recurrence. The area can be explored with the aid of an endoscope introduced into the nasal cavities through the nostrils, and in case of doubt, biopsies taken from all suspicious area. Our technique facilitates the repair of the surgical defect, and while not compromising the healing process it has a very low incidence of CSF leaks and infections. PMID- 9351992 TI - Micro-anatomical study of the carotid cave. AB - The surgical treatment of aneurysms located in the carotid cave is often hazardous and difficult. We studied the micro-anatomy of the carotid cave and its neighbourhood by microscopic observation and histological examination using 50 sides from 25 autopsy cases. The carotid caves were found in 34 out of the 50 sides (68%) examined and were usually located in the posteromedial aspect of the carotid dural ring. They were classified into three types according to the topographic micro-anatomy: the slit-type (17/50, 34%) which showed a small, thin recess of the dura mater with fine connective tissue loosely adhered to the carotid wall; the pocket-type (12/50, 24%) which had a definite dural pouch with the apex attached to the vessel wall; and the mesh-type (5/50, 10%) which formed a slit- or pocket-type dural cave covered with a mesh-like dural roof. The remaining 16 sides (32%) showed tight dural attachment without any caval structure around the dural ring. The posteromedial portion of the carotid dural ring had no contact with any bony structure, and this distinct anatomical feature thus appear to facilitate the formation of the carotid cave. Furthermore, the availability of this potential space and the closely situated origin of the superior hypophyseal artery as well as the haemodynamic effect of the internal carotid artery may allow the development of the carotid cave aneurysm. PMID- 9351993 TI - Comparative study of two customary cerebrospinal fluid shunting systems in early childhood hydrocephalus. AB - The validity of clinical studies on shunt-treated hydrocephalic patients is often hindered by inhomogeneity of the patient population examined, technical devices used, or by other specific factors. In an effort to introduce a homogeneous clinical study on hydrocephalic patients 66 hydrocephalic newborns and infants have been treated exclusively with CORDIS Orbis-Sigma Valve (OSV) System (CORDIS Corporation, Miami, USA) in 1990-1995. The results are compared with an equivalent group of 53 children treated with CODMAN Holter Valve (HV) System (CODMAN Inc., Randolph, USA) during a similar 5-year-period (e.g., 1986-1991). Searching for different reasons of shunt insufficiency in both groups demonstrates a more than double risk of shunt complication for ventriculo-atrial HV treated patients (VA-HV) in comparison with those treated ventriculo peritoneally with OSV System (VP-OSV): 4.22 versus 1.98 mean surgical procedures per person. The different revision and survival rates are discussed and specific problems are mentioned. PMID- 9351994 TI - Multiple intracerebral intravascular papillary endothelial hyperplasia. AB - Intravascular papillary endothelial hyperplasia (IPEH) is a rare benign reactive lesion usually found in thrombosed subcutaneous blood vessels. It uncommonly occurs in the central nervous system and may be mistaken for a more malignant type of tumour such as angiosarcoma. We present a first case of multiple IPEH occurring intracranially in a 51-year-old woman. She developed neurological compromises secondary to the mass affect of the haematoma arising from one of the lesions. Prompt surgical evacuation of the haematoma stabilized her condition. Surgical treatment, pathological findings, radiographic characteristics, and a review of the literature are presented. PMID- 9351995 TI - Intramedullary neurenteric cyst without any associated malformation. One case evaluated by RMI and electron microscopic study. AB - A 46 years old woman presented with several years history of low back pain. For five years she suffered from weakness of the left lower limb and three years later she experienced an episode of right foot weakness. She suffered too from occasional urinary urgency. The examination showed decreased power and diminished sensory perception in the left leg. On myelography, a block at L2 level was observed. RMI evaluation showed an intramedullary cyst in the anterior part of the spinal cord without any enhancement of its wall by the Gadolinium. At operation a thin-wall cyst was found containing clear fluid. After a biopsy of the wall, a cystosubarachnoid shunt was performed. Histological examination of the surgical sample showed a simple cuboidal epithelium lying on collagen fibrills. Electron microscopic studies showed ciliated cells with a clearly visible basement membrane. The diagnosis of neurenteric cyst was confirmed. In the postoperative course the patient complained about sensory loss of the legs and the perineal area. Six months later, she exhibited a sensory disturbance of the feet and the right sacral area, a motor deficit of the distal left leg without urinary disturbance. Neurenteric cysts are dysraphic lesions which can be observed without other abnormalities. They are usually extramedullary and the intramedullary forms are very rare: among 5 cases reported in the literature, one has been evaluated by RMI. In the absence of enhancement by the Gadolinium, the other possible diagnosis seems an ependymal cyst. Contrary to extramedullary forms the postoperative course of intramedullary neurenteric cysts are not always eventful. Because the cyst wall cannot be removed, repeated RMI are desirable in the follow-up. PMID- 9351996 TI - MRI detection of spontaneous rupture of a well differentiated pineal teratoma. PMID- 9351997 TI - Calvarial perforation caused by astrocytoma with extra-medullary growth. PMID- 9351998 TI - Worm control practices on sheep farms in Nyandarua District of Kenya. AB - A questionnaire investigation was used to examine anthelmintic usage and practical worm control for sheep on 50 farms selected randomly in Nyandarua District of Central Kenya. Control of helminths was based primarily on the use of anthelmintics on all 50 farms. On the majority (54%) of these properties, lambs were drenched two times per year. Ewes and rams were drenched three or four times per year on 74% of the farms. Most treatments were given at intervals of approximately 3 months with no specific drenching programmes. Anthelmintic doses for the sheep were based on weights estimated using visual appraisal on 98 and 96% of the properties for lambs and adult sheep, respectively. Only on a small proportion of the farms (22%) was the recommended weight of the heaviest animal used when drenching groups of either lambs or adult sheep. In 1994, the majority (68%) of farmers used levamisole (LEV) in combination with oxyclosanide (OXY) a fasciolicide, 10% used benzimidazoles (BZs), 10% LEV alone and 12% LEV and BZs together. This pattern of anthelmintics use was maintained from 1988 to 1994. Eighty one percent of the farmers had been using only LEV or BZs for three or more consecutive years from 1990 to 1994. The implications of these findings for the development of anthelmintic resistance are discussed. PMID- 9351999 TI - Natural Schistosoma mansoni infection in wild rats from Guadeloupe: parasitological and immunological aspects. AB - Rattus rattus is the predominant rodent in the mangrove area of Guadeloupe. Between 1990 and 1991 we found 73 R. rattus and five R. norvegicus. Among the infected rats with Schistosoma mansoni, 59% for R. rattus and 80% for R. norvegicus, the comparison of the median of the worm load was not statistically different. Both species of infected rats showed adult worms and eggs in the lungs and 20% of them showed, at the same time, two and even three generations of worms. Neither adults nor eggs were seen in the intestinal wall or stools of R. norvegicus, instead R. rattus had eggs in the liver, in the intestinal wall and the stools. Therefore, R. norvegicus gets infection as well as R. rattus, but does not participate in the transmission of the schistosomiasis. In order to elucidate this difference, we looked at the humoral recognition of these two rats, to the molecular antigens of the three stages of the parasite: cercaria, adult worm (AWA) and egg (SEA). In general, R. norvegicus recognized cercarial antigens more frequently than R. rattus, 73, 81 and 172 kDa being statistically different. Regarding AWA, molecules 82, 86, 117 and 150 kDa were recognized more often by R. rattus as compared to R. norvegicus. The reverse was true for the 18, 33 and 61 kDa. Only the differences between 61 and 150 kDa molecules were statistically significant. With respect to SEA, R. norvegicus recognized more 28, 45, 47, 49, 64 and 92 kDa molecules than R. rattus, but the latter recognized the 140 kDa molecules of SEA to a higher degree (95 and 140 kDa were significantly different). It is plausible that the immune response to cercarial invasion is more effective in R. norvegicus in allowing the parasites to reach adulthood, but it does not let them live in the mesenteric veins and therefore to lay their eggs in the intestinal wall and feces. PMID- 9352000 TI - Detection of lectin activity in Leishmania promastigotes and amastigotes. AB - Cell lysates from 16 strains of eight Leishmania species were used to test haemagglutination activity (HA) against a variety of RBC. HA was detected using native or neuraminidase-treated rabbit RBC; it was found in promastigotes of all the Leishmania strains tested and in axenic amastigotes of L. mexicana. The HA was trypsin-sensitive, heat-resistant and partially dependent on divalent cations. The HA was inhibited by amino-sugars, LPS from E. coli K 235, fetuin and heparin. The HA is probably located on the surface of promastigotes, as shown by the same sugar-binding specificity when live cells were used in inhibition tests. Leishmania promastigotes were agglutinated with neoglycoproteins NAc-glc-BSA and NAc-gal-BSA. This agglutination was blocked by galactosamine, glucosamine and sialic acid, but not by glcNAc or galNAc. The level of HA is increased in axenic amastigotes when compared to promastigotes. In general, HA was found at a higher titre in infective compared to uninfective strains of Leishmania. These results suggest that the haemagglutinin could play a role in the vertebrate phase of the parasite life cycle, possibly in macrophage attachment or invasion. PMID- 9352001 TI - T- and B-cell responses of malaria immune individuals to synthetic peptides corresponding to non-repeat sequences in the N-terminal region of the Plasmodium falciparum antigen Pf155/RESA. AB - While the C-terminal repeat region of Pf155/RESA, a Plasmodium falciparum vaccine candidate has been extensively studied for B- and T-cell reactivities, little is so far known about the non-repeat region in this respect. The present study aimed at investigating the non-repeat sequence 171-227 of Pf155/RESA for T- and B-cell epitopes. Eight overlapping peptides were synthesised and assayed for their ability to stimulate peripheral blood mononuclear cells obtained from P. falciparum-immune donors to proliferate and to induce secretion of interferon gamma (IFN-gamma) and/or interleukin 4 (IL-4) using the ELISPOT assay. The plasmas of the corresponding donors were tested for antibody reactivity with the same peptides in ELISA. The individual cellular responses to the different peptides varied and in general they were not correlated, emphasising the importance of including several parameters for T-cell activation. The most frequent T-cell responses (proliferation, IFN-gamma and/or IL-4) were seen with two partially overlapping peptides corresponding to the sequences 171-185 and 181 195 that induced responses in 71 and 62% of the donors, respectively. Although, the frequency of responders was high, the magnitude of the responses was generally low. Two overlapping peptides corresponding to the sequence 186-206 bound antibodies from a large number of plasma samples. IL-4 producing cells were frequently found in donors whose sera contained antibodies to the corresponding peptide. However, there was no absolute correlation and many donors having anti peptide antibodies could also be induced to produce IFN-gamma. In conclusion, the non-repeat region of Pf155/RESA contains several epitopes inducing functionally distinct T-cell responses. The sequence 171-206 was found to contain both B- and T-cell epitopes recognised by almost all individuals naturally primed to malaria. Thus, this sequence should be a useful tool in future immuno-epidemiological studies and/or for inclusion into a subunit vaccine against the asexual blood stages of the P. falciparum parasite. PMID- 9352002 TI - Maternal diagnosis and treatment of children's fever in an endemic malaria zone of Uganda: implications for the malaria control programme. AB - A mother's ability to suspect malaria in the presence of fever has important consequences for child survival in malaria-endemic areas. This paper presents results of a clinic-based study of mothers' abilities to suspect malaria in the event of recognizing fever and other physiological and behavioral changes associated with the disease. The study population consisted of all (439) women or mothers who had accompanied children 5 years and below to the Old Mulago Hospital, Kampala, Uganda over a 10 day period during the malaria season of 1992. The children were those who had fever as a major complaint at the time of the visit or those who had fever in the last 7 days and were visiting the clinic for the first time for the current illness. The children were physically examined and their blood tested for malaria parasites. Mothers' diagnosis was compared with clinical and laboratory diagnosis of malaria. Mothers associated the presence of fever with several types of illness and malaria was often not suspected. Only 40% of the mothers suspected malaria in their children. The mothers were poor at recognizing malaria when, in fact, it was present. The sensitivity of the mothers' diagnosis of malaria was found to be 37%; 63% of malaria cases were misclassified as other conditions. The doctors classified most (92%) of the cases presenting with fever as having malaria, but laboratory tests indicated that only 64% of the children really had malaria. The sensitivity of clinical diagnosis was 98%, but the specificity was only 18%. Ninety percent of the mothers gave some medicines before visiting the health centre; and, of these, 76% gave modern drugs exclusively, including antimalarials, antipyretics, antibiotics and other drugs. Among the modern drugs given to children suspected of having malaria, 50% were antimalarials. The most commonly used antimalarial was chloroquine tablets. Mothers indiscriminately administered antimalarials to children irrespective of the perceived cause of the fever. There is need to educate mothers to suspect malaria first in every case of febrile illness, just like the doctors do, and about the first line drugs for the treatment of malaria. PMID- 9352003 TI - Experimental cutaneous leishmaniasis. IV. The humoral response of Cebus apella (Primates: Cebidae) to infections of Leishmania (Leishmania) amazonensis, L. (Viannia) lainsoni and L. (V.) braziliensis using the direct agglutination test. AB - The direct agglutination test (DAT) was used to evaluate the serological response of 150 serum samples taken from 15 captive-bred capuchin monkeys Cebus apella. These animals had been experimentally infected with either L. (Leishmania) amazonensis, L. (Viannia) lainsoni or L. (V.) braziliensis. Monkeys infected with L. (L.) amazonensis or L. (V.) lainsoni were challenged with the homologous parasite one month after their spontaneous cure. DAT antigens were prepared from L. (L.) donovani, L. (L.) amazonensis and L. (V.) braziliensis. Antigens were difficult to standardise and it was impossible to produce an L. (V.) lainsoni antigen as parasites remained aggregated even after trypsinization. The DAT detected significant humoral responses in all the infected monkeys. Titres were higher when homologous antigens were used, especially in secondary responses. This suggests that homologous antigen should be used to detect antibodies in human cutaneous leishmaniasis. PMID- 9352004 TI - An evaluation of Schistosoma japonicum infections in three villages in the Dongting lake region of China. I. Prevalence, intensity and morbidity before the implementation of adequate control strategies. AB - We examined three Chinese villages (one farming village and two fishing villages) in an area highly endemic for schistosomiasis japonica in order to study the prevalence, intensity of infection and the associated morbidities before the implementation of adequate control strategies. Socio-economic status, medical histories including the frequency and type of water contact, physical examinations, parasitological examinations and questionnaires relevant to the knowledge of schistosomiasis were performed on a random sample of 1542 individuals (45% female; 55% male). The prevalence of Schistosoma japonicum was 9.4% in the farming village and 16.5 and 26.2% in the fishing villages. Eighty three percent of the infected population had light infections (8-100 eggs per gram stool (epg)) and only 6% had heavy infections (> 400 epg). Both the prevalence and intensity of infection varied significantly (P < 0.01) with the frequency of water contact. All the morbidity indicators (weakness, inability to work, diarrhoea, hepatomegaly and splenomegaly) were significantly higher (P < 0.01) among those infected with S. japonicum. Knowledge of schistosomiasis, in general, was unsatisfactory in all three villages; 12.4% of the population was infected when their knowledge of schistosomiasis was good, whereas 26.6% of the population was infected when their knowledge was poor. Further, it appears that schistosomiasis control based on selective chemotherapy (praziquantel) of randomly selected stool-positive individuals was ineffective in significantly reducing the prevalence of S. japonicum and its associated clinical manifestations in the villages under study. PMID- 9352005 TI - An examination of current control strategies for Asian schistosomiasis in the Dongting lake region of China. II. A five year follow-up survey on Qingshan island. AB - In 1995-1996 we conducted an epidemiological survey in two communities (1656 individuals) on Qingshan island, Hunan province P.R. China, in order to determine the efficacy of current control strategies since their upgrading in 1991. In 1996, the overall prevalence for Schistosoma japonicum, Ascaris lumbricoides, Ancylostoma duodenale, and Trichuris trichiura had decreased moderately since 1991. The age-specific prevalence for S. japonicum for each of the representative age groups decreased slightly, but there was a significant reduction in these prevalences for the 5-9 (P < 0.01), 55-59 (P < 0.05) and the over 60 (P < 0.01) age groups. The 1996 intensities of infection for schistosomiasis were higher for all the age categories except for those aged 0-4 and 25-29 years of age. When the study population was further classified according to the percent uninfected, lightly infected (8-100 eggs/g (epg)), moderately infected (101-400 epg) and heavily infected (> 400 epg) for S. japonicum, there were fewer (5.6%) people infected in 1996 but the proportions of moderately (21.3 vs. 15.5%) and heavily (7.6 vs. 2.3%) infected individuals were higher than those observed in 1991. The reported cases of weakness and hepatomegaly (MSL > or = 3) were significantly lower (P < 0.01) in 1996 for both uninfected and infected (all intensities) individuals. General episodes of diarrhoea were also significantly lower in 1996 for those lightly (P < 0.05) and heavily (P < 0.01) infected. Likewise, the occurrence of splenomegaly (Hackett's > or = 2) was significantly lower among uninfected (P < 0.01) and heavily infected (P < 0.05) patients. In summary, although significant progress has been made in controlling schistosomiasis and other helminth infections in this highly endemic focus for schistosomiasis, there is still room for improvement. Chemotherapy for bovines and humans, mollusciding for Oncomelania control and health education should be initiated and upgraded if the health and well being of these island communities is to further improve. PMID- 9352006 TI - Direct PCR amplification and sequence analysis of extrachromosomal Plasmodium DNA from dried blood spots. AB - The Plasmodium parasite possesses two extrachromosomal genomes; the mitochondrial genetic element and the extrachromosomal plastid-like DNA. The latter has only been fully described for one culture strain of P. falciparum. In this study, a rapid procedure for amplifying plastid DNA from dried blood spots of blood infected with different malaria species was developed. PCR amplification of a 595 bp fragment within the plastid-like large subunit ribosomal-RNA (LSU-rRNA) gene was achieved using primers derived from the P. falciparum sequence. The PCR product was observed in all Plasmodium species examined. Sequence analysis of amplified products homologous to an LSU-rRNA fragment of the plastid-like extrachromosomal circle revealed extensive conservation between Plasmodium species including P. falciparum, P. vivax, P. malariae and P. berghei. PMID- 9352007 TI - Canine filarial infections in north Taiwan. AB - To assess the current status of Dirofilaria immitis infection and to determine whether there were other canine filarial infections in north Taiwan, postmortem examination was conducted in 180 stray dogs more than 12 months old. Blood and serum samples were examined using a modified Knott's test and an antigen detecting enzyme-linked immunosorbent assay (ELISA) kit, respectively. Filarial infection was found in 60.6% of the dogs: 55% with D. immitis and 12.2% Dipetalonema reconditum. Moreover, the ELISA was determined to be more sensitive than the Knott's test. Although canine heartworm infection in Taiwan has been attributed to the unrestricted import of dogs from endemic areas, the results of this study indicate that transmission of D. immitis and Dip. reconditum may occur indigenously in the local canine population. This study is also the first record of Dip. reconditum in Taiwan. PMID- 9352008 TI - Visceral leishmaniasis from Bal'a, Palestine, caused by Leishmania donovani s.1. identified through polymerase chain reaction and restriction fragment length polymorphism analysis. AB - A 5-year old female from Bal'a, Tulkarm area, Palestine, was admitted with an 8 month history of fever, excessive night sweating, abdominal distension and enlargement, weight loss and sever anorexia. She was investigated elsewhere without reaching specific diagnosis. On admission, the history and symptoms were compatible with visceral leishmaniasis and bone marrow aspirate was positive for Leishmania amastigotes. The serum titer, using IFAT, was 1:640 for L. infantum and 1:320 for L. major promastigotes. When bone marrow material was also subjected to PCR followed by RFLP enzyme analysis, three fragments of the PCR product of the parasite present were obtained: two fragments of 260 bp and one fragment of 80 bp, identical with the pattern obtained with L. donovani. The patient received sodium stibogluconate, 200 mg IM for 30 days. Six months after treatment, the spleen was 2 cm below the costal margin, the liver was not palpable and she gained 1 kg. This case alerts general practitioners, pediatricians and health authorities to the presence of visceral leishmaniasis in Palestine and to the possibility of the disease being encountered in Jordan. PMID- 9352009 TI - Expression of IGFs and their receptors is a potential marker for embryo quality. AB - PROBLEM: Insulin-like growth factors (IGFs) and insulin have been demonstrated to stimulate oocyte maturation and embryo development. Therefore, the expression of IGFs and their receptors may be an important intrinsic factor for embryo growth and may be a potential marker for embryo quality. METHOD OF STUDY: Thirty donated day 3 embryos were cultured in vitro for an additional 3 days to observe their developmental potential and were semiquantitatively analyzed for the expression of IGF-I, IGF-II, IGF-IR, IGF-IIR, and insulin-R. RESULTS: Our results show that the activity of these gene expressions correlates well with the morphological assessment and that high and more gene expressions were often associated with embryos of high growth potential. CONCLUSION: The IGF system may indeed play an important role in human embryogenesis; IGF gene expressions can be a good indicator of embryonic developmental stage and/or growth potential; finally, the IGF system can serve as a marker for embryo quality. PMID- 9352010 TI - Regulation of decidual cell and chorion cell production of interleukin-10 by purified bacterial products. AB - PROBLEM: To determine whether cultured human decidual cells and chorion cells produce interleukin-10 (IL-10) after incubation with purified bacterial products. METHOD OF STUDY: Decidual cell cultures and chorion cell cultures were established by standard techniques. With confluence, monolayers of each culture were incubated with purified bacterial products, including various concentrations of lipopolysaccharide (LPS), lipid A, and lipoteichoic acid (LTA) for 16 hr in quadruplicate. Culture supernatants were collected and assayed for immunodetectable IL-10 by enzyme-linked immunoadsorbent assay (ELISA). RESULTS: Both decidual cell cultures and chorion cell cultures produced significant quantities of IL-10 after stimulation with LPS, lipid A, and LTA. Cultures of decidual cells produced more IL-10 than did chorion cell cultures. CONCLUSIONS: Our data indicate that both maternal decidual cells and fetally derived chorion cells can produce IL-10 after incubation with bacterial virulence factors. This finding contrasts with our previous findings in which chorion cells did not produce IL-10 after stimulation with IL-1 beta, suggesting that chorion cell production after incubation with bacterial products is independent of IL-1 beta. We speculate that the contribution of anti-inflammatory IL-10 production by human gestational tissues to the inflammatory process in these tissues may be overcome or abrogated by the pro-inflammatory process. PMID- 9352011 TI - Detection of human defensins in the placenta. AB - PROBLEM: The placenta is a highly selective barrier against the hematogenous dissemination of infectious agents. Despite the presence of seemingly intact physical and immunologic barriers, infections nonetheless occur. These observations prompted the examination of placental tissue, amnion, and chorion for previously unrecognized protective mechanisms. METHOD OF STUDY: Messenger RNA from term placenta, amnion, and chorion were reverse transcribed using a 3' RACE adapter. 3' rapid amplification of cDNA ends (RACE)-polymerase chain reaction (PCR) was conducted on cDNA from these tissues to detect the presence of human defensins. Southern analysis and partial sequence analysis were subsequently performed to confirm identity. RESULTS: PCR amplification of placental, amnion, and chorion cDNA yielded a 468-bp product and a weakly detectable band of 300 bp. Southern analysis demonstrated two corresponding hybridizing bands in the placenta, amnion, and chorion but not from a negative cDNA control. Partial sequence analysis of the 468-bp product from placenta confirmed the presence of either defensin 1 or 3 in human placenta. CONCLUSIONS: The human placenta, amnion, and chorion express defensins at the level of transcription. These findings suggest that a novel and previously unrecognized mechanism of protecting the fetus against infection may be present within these tissues. PMID- 9352012 TI - Soluble human leukocyte antigens, interleukin-6, and interferon-gamma during pregnancy. AB - PROBLEM: Soluble human leukocyte antigens (sHLA), interferon-gamma (IFN-gamma), and interleukin-6 (IL-6) were studied during human pregnancy to test the hypothesis that sHLA concentrations are regulated by these specific cytokines. METHOD OF STUDY: Enzyme-linked immunoadsorbent assays (ELISA) were used to measure sHLA I and II in maternal circulation, cord blood, and placenta effluents of pregnant and nonpregnant women; maternal serum cytokines were also determined. RESULTS: sHLA in maternal and cord blood were equivalent to that in the placenta. By the third trimester, sHLA I concentrations in maternal plasma were significantly reduced compared to the first or second trimesters. sHLA II was increased during the second trimester relative to that postpartum. Maternal IL-6 and IFN-gamma concentrations were not statistically different throughout gestation or postpartum. CONCLUSIONS: These data do not suggest a role for maternal plasma IL-6 or IFN-gamma in regulation of systemic sHLA class I during pregnancy, but they do not address whether such events take place in local tissues of the maternal-fetal unit. PMID- 9352013 TI - Protein transport across the in vitro perfused human placenta. AB - PROBLEM: Placental transport of various proteins present in human serum, such as immunoglobulins (IgG, IgA), specific anti-tetanus IgG (anti-TT-IgG), and tetanus toxoid-antigen (TT-AG), was investigated. In addition, the transport of IgG modified with biotin (IgG-BT) and 14C-bovine serum albumin (14C-BSA, a permeability marker for macromolecules), was assessed. METHOD OF STUDY: During the perfusion of an isolated cotyledon from human term placenta the perfusate was recirculated on both maternal and fetal sides. After an initial stabilisation phase of 2 hr (control phase), media on both sides were exchanged and perfusion was continued comparing two different conditions (experimental phase). In the first group (control experiments [A, n = 3]), no test proteins were added during the experimental phase (4-6 hr). In the second group (B, n = 5), during the experimental phase (6 hr) the maternal perfusion medium contained IgG (Sandoglobuline, 6-10 g/L), anti-TT-IgG (21-25 mg/L), TT-AG (0.19-0.24 mg/L), and IgA (0.13-0.19 g/L). IgG-BT (2 g/L) and 14C-BSA (30-40 nCi/ml) were added to the medium on the maternal side. IgGs and TT-AG were determined by specific enzyme linked immunosorbent assay. RESULTS: Both groups showed stable metabolic conditions with constant rates of glucose consumption, lactate production, and hormone (human chorionic gonadotropin, human placental lactogen) release observed throughout the experiment. Washout levels of endogenous IgG and IgA observed in the maternal circuit at the end of the control period were 5 and 1000 times higher than in the fetal circuit. In the experimental phase these levels remained constant at 50-80% of control levels with no change in the last 4 hr of perfusion (group A). In group B, with addition of extra proteins, trace amounts of IgG-BT, IgA, and 14C-BSA were detectable in the fetal circuit within 1 hr, with no significant further increase in circulating levels in the following 4 hr of the perfusion. In contrast, the detection of IgGs in the fetal circuit was delayed by 2 hr; thereafter, a continuous linear increase was observed for all IgGs. TT-AG in fetal perfusate was below the detection limit. TT-AG was found on the fetal side only after ultrafiltration of samples obtained at the end of the experiment. For permeability comparison, the ratio between concentrations on the fetal and maternal side multiplied by 100 ([F:M] x 100), as detected after 6 hr of perfusion, was assessed (n = 5, mean +/- SD). Labelling of IgG with biotin (IgG BT) reduced its placental transfer by a factor 10 (0.04 +/- 0.01) when compared with the natural IgG (0.49 +/- 0.08) or the specific antibody (anti-TT-IgG). The relative fetal-to-maternal ratio found for TT-AG (0.48 +/- 0.12) was similar to anti-TT-IgG (0.46 +/- 0.11), and approximately 4 and 50 times that of 14C-BSA (0.12 +/- 0.03) and IgA (0.01 +/- 0.01), respectively. Considering that the molecular weights of TT-AG and anti-TT-IgG were at least twice that of BSA and similar to IgA, the difference in transfer suggests a specific mechanism of transport. CONCLUSIONS: Compared with other proteins there is a significantly increased transfer of IgGs across the in vitro perfused human placenta from the maternal to the fetal side, indicating a specific transport mechanism. The similarity in transfer of anti-TT-IgG and tetanus antigen may suggest the transport as antibody-antigen complex. PMID- 9352015 TI - The effect of interleukin-1 beta and interleukin-4 on the expression of prostaglandin receptors EP1 and EP3 in amnion WISH cells. AB - PROBLEM: Although prostaglandin E2 (PGE2) is believed to modulate biochemical and immunological events leading to parturition, the role of prostaglandin E receptors during labor has not been investigated. METHOD OF STUDY: Amnion WISH cells were incubated in media containing increasing concentrations of either interleukin-1 beta (IL-1 beta) or IL-4. Increased EP1 and EP3 protein expression was determined by Western blot analysis with peptide-specific antibodies. Concomitant measurements of culture media PGE2 were made by an enzyme immunoassay. RESULTS: Incubation of WISH cells with IL-1 beta or IL-4 caused a two- to three-fold increase in EP1 protein levels. IL-1 beta and IL-4 also caused six- and two-fold increases, respectively, in culture fluid PGE2 concentrations. IL-1 beta or IL-4 had no effect on EP3 protein levels. CONCLUSIONS: Based on these results, it is proposed that IL-1 beta and IL-4 may be involved in the initiation and promotion of labor by inducing EP1 levels and PGE2 production in amnion. PMID- 9352014 TI - Regulation of interleukin (IL)-6 and IL-8 production in an amnion-derived cell line by cytokines, growth factors, glucocorticoids, and phorbol esters. AB - PROBLEM: To determine whether amnion cells produce interleukin (IL)-6 and -8 and thus may contribute to the high concentrations of these cytokines in amniotic fluid at term. METHOD OF STUDY: Amnion-derived WISH cells were treated in culture with stimuli over 16 hr, and IL-6 and IL-8 concentrations in the conditioned media were measured by enzyme-linked immunosorbent assay or bioassay (IL-6 only). RESULTS: IL-8 production was approximately 5-fold higher than that of IL-6 under basal and stimulated conditions. Significant (by Dunnett's test after analysis of variance) stimulation of production of both cytokines was achieved by IL-1 beta (> 0.2 ng/ml), TNF alpha (> 10 ng/ml), and the phorbol ester, phorbol 12 myristate 13-acetate (> 2 nM), over a 16-hr culture period. Epidermal growth factor at 10 ng/ml induced a small increase in production of IL-8, but not of IL 6, whereas bacterial lipopolysaccharide had minimal effects on production of either cytokine. Basal and cytokine-stimulated IL-6 and IL-8 production was inhibited by dexamethasone at concentrations equal to or greater than 1 nM. CONCLUSION: These findings suggest that amnion may be a significant contributor to the IL-6 and IL-8 content of amniotic fluid, and that WISH cells may be a suitable model for the study of cytokine production by amnion epithelial cells. PMID- 9352016 TI - Amniotic fluid granulocyte colony stimulating factor levels: a rapid marker for diagnosing chorioamnionitis. AB - PROBLEM: To assess the usefulness of amniotic fluid (AF) granulocyte colony stimulating factor (G-CSF) levels as a rapid marker for diagnosing chorioamnionitis. METHOD OF STUDY: AF levels were obtained from term and preterm patients with and without chorioamnionitis (CAM). Patients with urinary tract, respiratory tract, and other infections were excluded. Results obtained from the AF G-CSF assays were compared with those from other parameters used for diagnosing CAM: maternal fever, leukocytosis, tachycardia, fetal tachycardia, AF glucose levels, white blood cell count, Gram stain, and aerobic and anaerobic cultures. The sensitivity, specificity, and predictive values were calculated. RESULTS: In the uninfected AF samples, G-CSF levels were present but low, ranging from 400 to 1600 pg/ml. Levels in the infected samples, however, were markedly increased, ranging from 1600 to 14,000 pg/ml; P < 0.05. When a cutoff of 2000 pg/ml was used as a clear marker for CAM, the sensitivity was 67%, the specificity was 100%, and the positive and negative predictive values were 100% and 86%, respectively. The comparison of the other AF G-CSF laboratory parameters also revealed high sensitivity, specificity, and predictive values for detecting CAM. CONCLUSION: (i) AF G-CSF levels are elevated in CAM. (ii) An AF G-CSF level > 2000 pg/ ml is a strong positive predictor of CAM. (iii) Elevated AF G-CSF levels appear to be more reliable in predicting CAM than any other single test currently used in clinical practice. PMID- 9352017 TI - Immunolocalization of the inducible nitric oxide synthase isoform in human fetal membranes. AB - PROBLEM: Nitric oxide (NO) synthesized by fetal membranes may protect the fetus from maternal infection or immune challenge or have a tocolytic effect on myometrium. The sites of synthesis and enzymes responsible for NO production in human fetal membranes remain unidentified. METHOD OF STUDY: Fetal membranes were obtained from four groups of patients: term (> 37 weeks gestation) or preterm (< 37 weeks gestation), both either in labor or not in labor. Frozen sections of membrane rolls were immunostained for inducible (iNOS) and endothelial (eNOS) nitric oxide synthase isoforms and the monocyte/macrophage marker CD14. RESULTS: Positive iNOS immunostaining was found in fibroblasts of amnionic and chorionic mesenchyme and in decidual macrophages identified by CD14 from all four groups of tissues. No iNOS immunostaining was seen in amnion epithelium or chorion trophoblast. Very intense iNOS staining was seen with evidence of monocyte/macrophage invasion of membranes. eNOS immunostaining was only found in decidual vascular endothelium. CONCLUSIONS: Constitutive expression of iNOS in decidual macrophages and fetal membrane fibroblasts may form an immune barrier against maternal insult. In chorioamnionitis, macrophage recruitment and NO expression may be part of the maternal immune response. PMID- 9352018 TI - Expression of cell adhesion molecules in the extravillous trophoblast is altered in IUGR. AB - PROBLEM: The invasion of trophoblast cells into the uterine wall and its arterial system is essential for the normal development of pregnancy. Cell adhesion molecules (CAM), such as the immunoglobulin superfamily and integrins, play a crucial role in a number of immunological reactions and in the invasion of the human trophoblast. Intrauterine growth restriction (IUGR) has been associated with abnormal trophoblast invasion. Therefore, the expression of CAM in the extravillous trophoblast of pregnancies complicated by IUGR might be different from normal pregnancies. METHOD OF STUDY: Normal (n = 21) and IUGR (n = 19) placentas were collected and stored at -70 degrees C. Immunohistochemistry (avidin-biotin complex peroxidase-doublestaining) of frozen tissue sections was performed using antibodies specific for the immunoglobulin superfamily vascular adhesion molecule-1 (VCAM-1; CD 106), intercellular adhesion molecule (ICAM-1) (CD 54), ICAM-2 (CD 102), ICAM-3 (CD 50), the integrins alpha 2 beta 1, alpha 3 beta 1, alpha 4 beta 1, alpha 5 beta 1, alpha 6 beta 1 and cytokeratin. The percentage of immunopositive extravillous trophoblast cells (EVT) and the intensity of the immunoreactivity for the various CAM and integrin antibodies was assessed. RESULTS: In IUGR placentas, there was less expression of VCAM-1 (CD 106), alpha 2 beta 1, alpha 3 beta 1, and alpha 5 beta 1 (P < 0.05) in the extravillous trophoblast than in normal pregnancies. Finally we observed for the first time that ICAM-3 was expressed on EVT and that its expression was markedly up-regulated in the EVT or IUGR placentas. No differences were found for ICAM-1 (CD 54), ICAM-2 (CD 102), alpha 4 beta 1 and alpha 6 beta 1. CONCLUSION: Our data show that there are significant differences in the expression of cell adhesion molecules of the extravillous trophoblast from IUGR and normal pregnancies. These differences might reflect changes in the immunological reactions and cell-cell interactions between mother and the developing fetus which could interfere with fetal growth. PMID- 9352019 TI - Vascular endothelial growth factor is increased in patients with preeclampsia. AB - PROBLEM: This study was conducted to determine whether altered levels of vascular endothelial growth factor (VEGF) may play a role in the pathogenesis of preeclampsia. METHOD OF STUDY: Maternal plasma samples were collected from 19 patients with preeclampsia (group A) either before the onset of labor, or before induction of labor or medical intervention. Plasma samples were also obtained from 19 normotensive patients with uncomplicated pregnancies (group B), who were matched with the patients with preeclampsia for gestational age and parity. Samples were frozen at -70 degrees C until assayed for VEGF by a specific enzyme linked immunoassay. RESULTS: The mean maternal age was similar in groups A and B. For both groups the VEGF was detectable in all plasma samples. However, the plasma concentrations of VEGF were significantly increased in the group A patients, compared with those in group B (median, 47 ng/ml; range, 10.6-72 ng/ml versus median, 13.6 ng/ml; range, 0.66-20 ng/ml; P < 0.001). In group A, a positive correlation was noted between VEGF concentrations and the systolic and diastolic blood pressure (r = 0.56; P = 0.01 and r = 0.48; P = 0.037, respectively). CONCLUSIONS: Maternal plasma VEGF levels were elevated in the patients with preeclampsia and correlated with the severity of hypertension, suggesting a role for VEGF in the pathogenesis of preeclampsia. PMID- 9352020 TI - Amniotic fluid granulocyte colony stimulating factor levels in chorioamnionitis do not predict neonatal sepsis. AB - PROBLEM: To assess the usefulness of amniotic fluid (AF) granulocyte colony stimulating factor levels (G-CSF) in chorioamnionitis (CAM) to predict neonatal sepsis. METHOD OF STUDY: AF samples were obtained from term and preterm patients with (Group I) and without (Group II) CAM and were assayed for G-CSF levels. Patients with other infections were excluded. All AF samples were also tested for gram stain and cultures. The sensitivity, specificity, and predictive values of these parameters for diagnosing neonatal sepsis were assessed. RESULTS: Positive AF cultures were the best predictors of neonatal sepsis in CAM, with a sensitivity of 67% and a positive predictive value (PPV) of 80%. Elevated AF G CSF levels (> 1,000 pg/ml) were poor predictors of neonatal sepsis with a sensitivity of 29% and PPV of 39%. CONCLUSION: Even though AF G-CSF levels were markedly elevated in patients with CAM, they were poor predictors of subsequent neonatal sepsis. PMID- 9352021 TI - Lethal outcome of uterine infection in pregnant but not in nonpregnant rats and increased death rate with inhibition of nitric oxide. AB - PROBLEM: Limited information is available on potential differences in sensitivity to urogenital infections between pregnant and nonpregnant hosts. METHOD OF STUDY: In this study, we evaluated Escherichia coli infectious complications in pregnant and nonpregnant rats and the effect of nitric oxide (NO) inhibitor, NG-nitro-L arginine methyl ester (L-NAME), on the outcome of an experimental uterine infection. RESULTS: Of the infected pregnant animals, 31% were found dead in 24 48 hr. The death rate was increased 2-fold (66%) with L-NAME treatment. No deaths occurred in nonpregnant animals with or without L-NAME treatment. The rate of uterine infection in pregnant animals was about 10-fold higher than in nonpregnant animals. CONCLUSION: We propose that infectious complications of pregnancy may be related to gestation-dependent sensitivity to the pathogenic microorganism and the host NO status. PMID- 9352022 TI - Suppressive effect on lymphoproliferation in vitro by soluble annexin II released from isolated placental membranes. AB - PROBLEM: Syncytiotrophoblast microvillous plasma membranes (StMPM) are potent suppressors of lymphoproliferation in vitro. We have previously shown that soluble annexin II (AII) is present at higher levels in retroplacental serum (RPS) than in peripheral serum, and that soluble AII has an immunosuppressive effect. The aims of this study were to determine whether AII can be released from StMPM and whether soluble AII from StMPM exerts any immunosuppressive effect. METHOD OF STUDY: Isolated StMPM were incubated in growth medium for 18 hr and supernatants were prepared by ultracentrifugation. Soluble AII was detected by immunoblotting. StMPM, StMPM supernatant, and affinity-purified AII were analysed in a lymphoproliferation assay for immunomodulating activity. RESULTS: AII heavy chain and its p11 light chain were detected both in StMPM supernatant and in RPS after removal of StMPM particles by ultracentrifugation. StMPM, StMPM supernatant, and purified AII suppressed lymphoproliferation in a dose-dependent manner. Absorption of AII from StMPM supernatant reduced the suppressive activity. The suppressive effect of StMPM supernatant and purified AII was completely reversed by heating at 100 degrees C for 30 min or by adding recombinant interleukin-2 at 100 units/ml. Although StMPM and affinity-purified AII suppressed the proliferation of lymphocytes from all donors tested, StMPM supernatant suppressed the proliferation of lymphocytes from 12 of 23 donors. Six of eight female non-suppressed donors were multiparae, whereas five of five female suppressed donors were nulliparae. CONCLUSIONS: Annexin II is released by isolated placental membranes in vitro and is present in RPS, indicating in vivo release of AII at the fetomaternal interface, probably as AII heterotetramer. AII has immunosuppressive activity and may be important in fetal allograft survival. PMID- 9352023 TI - Paternal leukocytes selectively increase secretion of IL-4 in peripheral blood during normal pregnancies: demonstrated by a novel one-way MLC measuring cytokine secretion. AB - PROBLEM: It has been proposed that immune responses in normal pregnancy are Th2 like, thereby protecting the fetus and placenta from being rejected. Some studies have shown Th2-deviated systemic responses to different antigens and mitogens. The aim of this study was to demonstrate the specific T cell cytokine responses directed toward paternal histocompatibility leukocyte antigen (HLA), because this is the most prominent target for rejection of the feto-placental unit. METHOD OF STUDY: A novel one-way mixed leukocyte culture (MLC) combined with the detection of cytokine secretion with a sensitive ELISPOT assay was developed. Peripheral blood from 11 pregnant women was investigated with respect to allo-reactivity toward paternal leukocytes and pooled leukocytes from unrelated blood donors. This was done at three different occasions during pregnancy and 8 weeks after delivery. Nine age-matched non-pregnant women served as controls. RESULTS: In the second and third trimesters of pregnancy significantly larger numbers of IL-4 secreting cells (Th2) were induced by paternal leukocytes as compared to unrelated leukocytes. CONCLUSIONS: The findings indicate a selective immune deviation toward Th2, which may protect the fetus from rejection and thus may be an important homeostatic mechanism in normal pregnancies. PMID- 9352024 TI - Preliminary characterization of an immunosuppressive inducer factor secreted by the JEG-3 choriocarcinoma cell line: in vitro and in vivo studies. AB - PROBLEM: The direct immunosuppressive and suppressive inducing capacities of supernatants from human trophoblastic choriocarcinoma cell lines (HCS) are well investigated in several former studies. The responsible factor is not yet determined. METHOD OF STUDY: We first confirmed those data and we purified a 3-5 kDa suppressor-inducer factor from HCS by using high performance liquid chromatography (HPLC) on both DEAE and gel filtration columns, followed by ultrafiltration. We then tested the activities of such isolated fractions on in vitro immune responses from human cells and in vivo by its effects in a murine local graft-versus-host (GVH) assay (popliteal lymph node assay, PLN). RESULTS: A single fraction induces both "direct suppression" in vitro as well as in vitro suppressor cell activation/development in human peripheral blood lymphocyte cultures as assessed by suppression of cells cultured in such a fraction containing culture medium of the mixed lymphocyte reaction. Furthermore the very same fraction suppresses in vivo murine allogeneic immune responses as assessed by a local GVH reaction (PLN assay). CONCLUSIONS: We have isolated a suppressive fraction, whose activities suggest that it might be of interest not only in reproductive immunology, but also in transplantation systems. PMID- 9352025 TI - Stability of serum interleukin-10 levels during the menstrual cycle. AB - PROBLEM: Menstrual cycle-associated variability in the circulating levels of several cytokines can be a confounding factor in measurements of in vivo cytokine levels in clinical studies. Since pregnancy-associated increases in interleukin 10 (IL-10) levels are well documented, we have investigated the variability in serum levels of IL-10 in healthy women at different stages of the menstrual cycle to ascertain whether this is a problem in comparative studies of circulating IL 10 levels. METHOD OF STUDY: We obtained fifty-four successive serum samples at points in the menstrual cycles of 12 healthy fertile women, precisely timed by measurement of the luteinizing hormone surge, and measured the interleukin-10 levels. RESULTS: Levels of IL-10 in successive serum samples from each woman taken on days LH - 7 (that is seven days prior to LH surge), LH - 4, LH + 1, LH + 7, and LH + 10 showed that IL-10 does not vary in a systematic way during the menstrual cycle. CONCLUSION: These results validate the sampling of women in studies of IL-10 levels in various clinical situations and establish that these levels are not dependent on menstrual cycle dates. They also suggest that menstrual cycle-related changes in IL-1 are not mediated by IL-10. The rise in progesterone in the luteal phase of the menstrual cycle is not mirrored by a rise in the circulating IL-10 level, which implies either that the pregnancy associated rise is not related to progesterone or that it is only observed at the higher progesterone levels in pregnancy. PMID- 9352026 TI - Induction of autoimmune prostatitis using liposomes is associated to peritoneal cells activation. AB - PROBLEM: Study and characterization of rat peritoneal cells (PC) involved in the induction of autoimmune prostatitis after the intraperitoneal administration of native extract of accessory glands (RAG) associated with liposomes (RAGL). METHOD OF STUDY: Induction of the autoimmune response in normal recipients by transferring PC or adherent-PC loaded with RAGL (RAGL-PC), but not with PC loaded with empty liposomes (L-PC). Characterization of the morphology, the ultrastructure, and the phenotype of L-PC or RAGL-PC. Study of the respiratory burst by the nitroblue tetrazolium (NBT) reduction assay after stimulation with phorbol myristate acetate (PMA) in both L-PC and RAGL-PC. RESULTS: Liposomes attached to the cell surface of the M phi were observed by electron microscopy. FACS analyses showed a similar staining pattern with high expression of Ia molecules on L-PC and RAGL-PC compared with controls. PMA-stimulated L-PC or RAGL PC markedly reduced the NBT compared with controls. CONCLUSION: Our results suggest that the effective uptake of liposomes and the initial activation of PC together with a prolonged stimulatory effect help to disrupt the tolerance state. The present experimental model is an interesting approach to further characterize events associated with antigenic presentation when an autoimmune response is triggered. PMID- 9352027 TI - Flow cytometric analysis of leukocytes in the human female reproductive tract: comparison of fallopian tube, uterus, cervix, and vagina. AB - PROBLEM: The tissues of the human female reproductive tract (Fallopian tube, uterus, cervix, and vagina) may play different roles in the provision of mucosal immunity. The purpose of this study was to develop a uniform method suitable for quantitative comparison of the leukocytes from all these tissues. METHOD OF STUDY: Tissues, typically 0.5-1.0 g, were dispersed by enzyme treatment. A flow cytometric gating procedure based on CD45-positivity and low far-red autofluorescence permitted unfractionated, freshly dispersed cells to be phenotyped with respect to T lymphocytes, B lymphocytes, macrophages, and granulocytes. RESULTS: Reproductive tract tissues contain leukocytes that represent approximately 6-20% of the total number of cells, with the Fallopian tubes and uterus containing a higher proportion of leukocytes than the cervix and vagina. The uterine endometrium from post-menopausal women has fewer leukocytes than does uterine endometrium from pre-menopausal women. T lymphocytes are a major constituent (30-60%) of leukocytes from all tissues. The Fallopian tube contains granulocytes as another major constituent; granulocytes are significantly less numerous in the other tissues. All tissues contain B lymphocytes and macrophages as clearly detectable but minor components. CONCLUSIONS: Three-color flow cytometry is an appropriate method for quantitative comparison of leukocytes from the different tissues of the female reproductive tract, during all phases of the menstrual cycle and within post-menopausal samples. Results indicate that the tissues differ from each other, particularly with respect to the large number of granulocytes in the Fallopian tubes. PMID- 9352028 TI - Post-thymectomy murine experimental autoimmune oophoritis is associated with reduced natural killer cell activity. AB - PROBLEM: Natural killer (NK) cells can influence the immune response by secreting potent lymphokines. It has been suggested that NK cells have a suppressive action on B cells, and that impaired NK cell activity may play a role in some types of autoimmunity. NK cell abnormalities have been reported in women with premature ovarian failure. We therefore examined NK cell activity during the development of murine experimental autoimmune oophoritis, which serves as a model for autoimmune ovarian failure in women. METHOD OF STUDY: Neonatally thymectomized and sham operated C57B1/6 x A/J (B6A) mice were prepared and sacrificed at 4, 6, 8, and 10 weeks after surgery. Splenic NK cell activity was determined in groups of five or more mice by measuring the percent specific lysis of target YAC-1 lymphoma cells using a standard 4-hr chromium release cytotoxicity assay. The number of splenic NK cells in neonatally thymectomized and sham-operated animals was also compared using flow cytometry. In a subsequent experiment, interleukin 12 (IL-12; NK cell stimulating factor) was administered to neonatal mice before neonatal thymectomy. RESULTS: Neonatally thymectomized mice with associated autoimmune oophoritis had a 75% reduction in the number of splenic NK cells, and 50% or greater reduction in splenic NK cell activity at 4, 6, and 8 weeks after surgery. IL-12 treatment before neonatal thymectomy maintained NK cell activity and was shown to ameliorate the associated autoimmune oophoritis. CONCLUSION: Murine post thymectomy autoimmune oophoritis is associated with reduced NK cell number and impaired NK cell activity, and in these respects the model is similar to premature ovarian failure in women. Research to define the relationship between NK cell abnormalities and the mechanism of ovarian failure in this model might lend insight into the pathogenesis of premature ovarian failure in women. PMID- 9352029 TI - Effect of rat placental culture supernatants on cellular and humoral immune responses. AB - PROBLEM: To evaluate the effect of rat placental culture supernatants (PS) on spontaneous, mitogen- and alloantigen-induced lymphoproliferation, antibody synthesis regulation, and symmetric/asymmetric antibody ratio. METHOD OF STUDY: The effect of PS was determined: (a) on cell proliferation of murine hybridoma cells and on spontaneous or ConA-induced proliferation of murine and rat splenocytes by thymidine incorporation; (b) on rat or mouse cell-mediated cytotoxicity (CMC) by 51Cr release; and (c) on antibody synthesis by enzyme linked immunoadsorbent assay (ELISA). RESULTS: With 20% PS, hybridoma cell inhibition was 37% and that of splenocytes up to 60%, whereas it was 75 and 43%, respectively, in the presence of ConA. Despite marked cell death, hybridoma proliferation index increased significantly. There was a drop in total antidinitrophenylated (DNP) immunoglobulin G1 (IgG1) antibody production and an increase in asymmetric antibody percentage, correlating with placental supernatant concentration. CONCLUSIONS: Rat placental culture supernatants inhibit cell proliferation in all cases, diminish total antibody production, and increase the percentage of asymmetric antibodies by the hybridoma, and they increase antibody production by rat splenocytes. PMID- 9352030 TI - Recent advances in andrology research: physiopathology and clinical application to fertility and infertility. AB - Sperm maturation depends on androgen and is mediated by several unidentified epididymal factors: glycoproteins and metabolites affecting acrosomal stability and fertilizing potential of capacitated sperm. Several genes encoding human epididymis-specific proteins have been described. One of the cloned epididymal cDNA encodes a polypeptide designated as HE4 with an estimated molecular mass of 10,000. Leydig cells are rich in lipid droplets and display epithelioid features. These cells have a cord-like arrangement; the cords are formed by one or two closely apposed cells. In between these cells, labyrinthine or canalicular-like spaces are opened in wide perivascular spaces that improve cell secretion of hormones and facilitate their transport into the blood, as well as the traffic of fluids and metabolites. Coagulation and liquefaction in human semen plays an important role in the capacitation of semen. The liquefaction of semen is retarded by the powerful synthetic inhibitors of 6-amidino-2-naphtyl-p guanidinobenzoate dimethansulfonate. PMID- 9352031 TI - Relationship between etiological factors and total motile sperm count in 350 infertile patients. AB - The prevalence of different etiologic factors has been evaluated in 350 male patients consulting the same physician in an urban, ambulatory setting for primary or secondary infertility of more than 1 year. Environmental factors such as alcohol or drugs represented 12% of the etiologies, acquired diseases such as varicocele and prostatitis 40%, congenital diseases and primary testicular failure 16.2%, idiopathic cases 19.4%, and abnormality of sperm transport 7.4%. The severity of sperm alterations in the different etiologic categories was evaluated by the total motile sperm count per ejaculate (TMS) (normal > 16). The TMS was less than 5 in classical causes of male infertility such as testicular failure, endocrinopathy, cancer, or antisperm antibodies. It was more than 10 in controversial causes of infertility such as varicocele, prostatis, chlamydial infections, and professional exposure to heat. After treatment, there was a nonsignificant increase of the TMS in the latter cases. In cases of azoospermia of pituitary origin, the TMS was normalized by a hormonal treatment. In some cases of azoospermia of possible obstructive origin, sperm appeared in the ejaculate after diclofenac treatment. The utility of andrological investigation and treatment is discussed. PMID- 9352032 TI - Effects of various concentrations of native seminal plasma in cryoprotectant on viability of human sperm. AB - To investigate the effect of native seminal plasma on the recovery of frozen human sperm, various concentrations of seminal plasma (0, 25, 50, 75, and 100%) were used in cryoprotectant for freezing sperm, and the viabilities of frozen thawed sperm were compared. The post-thaw sperm motility of 50 or 75% seminal plasma in the fertile group was significantly higher than that of 0, 25, or 100%. The post-thaw motility of 75% donor seminal plasma in the patient group was higher than that of other concentrations. It was suggested that a certain concentration of native seminal plasma in cryoprotectant would be helpful to the viability of human sperm cryopreservation. PMID- 9352033 TI - Fluorescence in situ hybridization studies on the sex chromosome constitution of human sperm. AB - More males are conceived than females and more males are born as a result of in vitro fertilization (IVF) and donor insemination procedures. All donor sperm samples are frozen for a minimum of 6 months before they are used. The ratio of more boys to girls has been consistently reported over the years. A similar finding has been noted in the Galway Fertility Unit at University College Hospital Galway, Ireland. Traditionally, it has been accepted that the reason for this excess is that a Y chromosome-bearing sperm swims faster than the X-bearing sperm because it contains less DNA and is therefore "lighter." To test this hypothesis, semen samples were collected at the Galway Fertility Unit from men presenting for routine semen assessments. Each sample was divided into a number of aliquots. The first aliquot was assessed as the "raw" ejaculate to measure the initial ratio of X to Y. The second aliquot was prepared using Percoll density gradients, which allows for greater recovery of sperm with higher motility and improved sperm function. The final aliquot was frozen. The frozen sample was later thawed and prepared using Percoll. The prepared sperm were kept for 48 h and sampled at the time of preparation and at 24 h and 48 h to establish if there was any differential survival over time. The X:Y ratio was analyzed using the technique of fluorescence in situ hybridization (FISH). This allowed the sex chromosomes to be specifically stained and identified simultaneously. No difference was found in the X:Y ratio of the sperm. Therefore, any selection for the Y sperm must take place at some later stage. PMID- 9352034 TI - Sperm motility and ATP content in seminal hyperviscosity. AB - Objective spermatic motility (Hamilton Thorne Research), the rapid progressive spermatozoa (grade A) recovery after swim-up, and the spermatozoa ATP content (bioluminescence) were studied in normoviscous and hyperviscous asthenospermic samples. The amplitude of lateral head displacement (ALH) was significantly lower in hyperviscous semen (normal: 4.6 +/- 0.7 microns [n = 20], high: 3.5 +/- 1.2 microns [n = 16]; p < .05). The grade A recovery percentage after swim-up was significantly higher in semens with high consistency (normal: 71.0 +/- 38.0 [n = 14], high: 181.3 +/- 108.9 [n = 6]; p < .05). The ATP content per living spermatozoa was in the normal consistency group 449.4 +/- 65.1 pmol per million living spermatozoa (n = 29) and in the high consistency batch 605.1 +/- 242.8 (n = 9), p < .05. In asthenospermia, the spermatozoa from hyperviscous samples have minor ALH values, better response to swim-up, and high ATP content than those from normoviscous ejaculates. PMID- 9352035 TI - Cryptorchidism: treatment with human chorionic gonadotropin--a Venezuelan experience. AB - The study comprised 323 cryptorchidic boys, between 6 months and 14 years of age (mean age 5.68 years) with 440 maldescended testes. Testicular position was graded as inguinal low or prescrotal (I), inguinal middle (II), inguinal high (III), and abdominal testes (IV). Boys before 4 years of age received human chorionic gonadotropin (hCG) as intramuscular injections (I.M.), 500 IU twice a week for 5 weeks; and boys 4 or more years of age received hCG (IM), 1000 IU twice a week for 5 weeks. The objectives of this study were to evaluate the response of maldescended testes to treatment with hCG, and to investigate possible associations between the patients' ages and position of the testes with the response to hCG. Out of the 440 maldescended testes, 329 were in an inguinal location (75%) and 111 were abdominally located (25%). The overall response to hCG was 40%, and the inguinal testes response was 49%, with the highest success rates (72%) for the prescrotal testes. A positive correlation was found (p < .0001) between the rate of success and the testicular position. There was no association between the hCG response and the age at which treatment was initiated. PMID- 9352036 TI - Evaluation of semen parameters in man with hyperprolactinemia induced by metoclopramide. AB - Hyperprolactinemia in man decreases libido and potency, but the few reports concerning its influence on spermatogenesis are contradictory. The aim of this study was to evaluate the effect of induced hyperprolactimemia on semen parameters. A total of 15 potentially fertile male volunteers, aged 28.2 +/- 4.3 years, were given 10 mg metoclopramide three times daily for 12 weeks. Serum and seminal plasma prolactin levels and semen parameters were determined before and 4, 8, and 12 weeks following initiation of metoclopramide administration. A fivefold increase of serum prolactin levels was observed, semen volume and abnormal sperm forms decreased, while spermatozoa velocity increased. On the contrary, no influence was noted on the number of spermatozoa per milliliter, the total number of spermatozoa, the percentage of motile spermatozoa, or the index of motility. Hyperprolactinemia seems to improve spermatozoal velocity and morphology, although direct effect of metoclopramide on these parameters cannot be excluded. PMID- 9352037 TI - Transtrochanteric rotational osteotomy for osteonecrosis of the femoral head. 43 patients followed for at least 3 years. AB - We reviewed 48 hips in 43 patients 3-7.1 years (average 4.6 years) after Sugioka transtrochanteric rotational osteotomy for osteonecrosis of the femoral head. The average age at operation was 41 years. Thirty-four patients were men and 9 women. Overall results at the final follow-up were satisfactory in 30 hips (62%). Kaplan Meier's survivorship was 62% at 3 years and 60% at 5 years postoperatively. Six hips for which the ratio of the intact area of the articular surface on the preoperative lateral radiograph was less than 30% showed further collapse. Five hips were converted to bipolar hemiarthroplasties or total hip arthroplasties. Complications, such as varus deformity, subtrochanteric fracture, and ectopic bone formation, occurred in eight hips. Five of them were operated on in the first 2 years of this series. Three of these five operations had unsatisfactory results. We conclude that satisfactory results can be achieved using this osteotomy by maintaining exact surgical technique and by limiting the surgical indications to hips with an intact area of more than one-third of the entire articular surface on the lateral radiograph of the femoral head. PMID- 9352038 TI - Histological and biomechanical observations of the rabbit patellar tendon after removal of its central one-third. AB - Using 35 Japanese white rabbits, a study was made of tissue regeneration and the mechanical properties of the patellar tendon after removal of its central one third. After removal of the central one-third of the patellar tendon on one side, in experiment 1 the strength of the entire patellar tendon including the regenerated tissue was compared with that of the patellar tendon on the opposite side with the central one-third removed at the time of killing, and in experiment 2 the strength of only the regenerated tissue was compared with that of the patellar tendon on the opposite side with two-thirds of the medial and lateral sides removed at the time of death. In experiment 1, the maximum load showed no significant difference between the operated side and the control. In one half of the cases, the strength of the operated side including the regenerated tissue was weak, suggesting weakening of the patellar tendon on the residual bilateral sides. In experiment 2, the maximum load of the regenerated tissue was significantly lower than that of the control, the former being 25% of the latter even at 6 months. Histologically, the characteristics of the cells and collagen fibers gradually approached those of normal tissue, but the crimp pattern of the collagen fibers and fibrils was evidently smaller than that of the control. These results indicate that regenerated tissue was still mechanically weak and immature at 6 months. PMID- 9352039 TI - Surgical treatment for myeloma of the bone. A retrospective analysis of 22 cases. AB - In a retrospective study, 22 patients treated surgically for solitary or multiple myeloma between 1980 and 1993 were analysed. The main complaint was pain. A fracture was observed in 7 cases and motor-sensory impaired neurology due to spinal compression in 3. Apart from incisional biopsies, tumour resections, reductions (with and without stabilization by osteosynthesis) and endoprotheses were performed either at the extremities or on the spine. In addition, radiation and chemotherapy were included in the therapeutical concept. Early mobilization was achieved in all cases, and the 5-year survival rate (Kaplan-Meier method) was 48%. The results presented in this study demonstrate that a variety of surgical interventions can be of importance in the treatment of myeloma of the bone, ranging from biopsy or even curative resections in selected cases to endoprosthetic replacement. Thus, good functional results can be achieved and maintained over often long survival times. PMID- 9352041 TI - Surgical treatment of bone sarcomas of the fibula. Analysis of 19 cases. AB - Nine patients with Ewing's sarcomas and seven patients with osteosarcoma of the fibula were treated surgically. The bone defect after tumour resection ranged from 5 to 25 cm (median 14 cm). Ten sarcomas were located in the proximal and six in the diaphyseal or distal fibula. Nine of ten patients with sarcomas located in the proximal fibula underwent a resection of the tumour including the common peroneal nerve. In one patient with a tumour in the proximal fibula, the peroneal nerve was preserved; however, this patient underwent amputation because of surgery with an intralesional margin. In five patients with a tumour in the distal fibula, the peroneal nerve was preserved. However, two of these five patients underwent amputation as an adequate surgical margin could not be achieved during resection. All ten patients in whom the peroneal nerve was resected achieved satisfactory function by wearing a peroneal brace. In patients with Ewing's sarcoma of the proximal fibula, preservation of the common peroneal nerve may be chosen as an alternative possibility of resection. PMID- 9352043 TI - Patient's memory or repeated pain and function scores as index for major clinical change caused by knee replacement? AB - Change of the clinical condition is the aim of replacement of the knee. This has at least four components: a clinical phenomenon, an event for the phenomenon, size of the event and time for the event. This study dealt with major change of pain and function at the time of remission. The agreement between the patients's opinion of change and a clinical change score was poor. That a special index of changes is needed is shown in this study. PMID- 9352042 TI - Application of extracorporeal shock-waves in the treatment of pseudarthrosis of the lower extremity. Preliminary results. AB - Between January 1991 and January 1996, pseudarthroses of the legs were treated prospectively in 48 patients by application of high-energy extracorporeal shock waves with an experimental device. The mean duration of pseudarthrosis was 12 months. On average, 2.4 surgical interventions had previously been performed. A total of 3000 impulses with an energy density of 0.6 mJ/mm2 was applied to the pseudarthrosis. Bony union was achieved in 60.4% of our patients after an average of 3.4 months. Failures were found especially in the atrophic types of pseudarthrosis as well as in congenital bone disorders like fibrous dysplasia or osteogenesis imperfecta. No serious complications were observed. Even after numerous surgical interventions high-energy extracorporeal shock-wave therapy showed a fair success rate. A higher success rate of this non-invasive method for the treatment of bony non-unions may be expected by applying strict selection criteria. PMID- 9352040 TI - Bone and muscle mass after femoral neck fracture. A controlled quantitative computed tomography study of osteosynthesis versus primary total hip arthroplasty. AB - The cortical bone mineral density (BMD), bone volume, bone mass and muscle volume of the thigh, and the BMD of the distal femur and proximal tibia were measured quantified by quantitative computed tomography (QCT) after an operation for a displaced femoral neck fracture. Twenty patients were randomized to osteosynthesis or total hip arthroplasty (THA). Both legs were scanned after 18 months, and the operated side was compared with the healthy side. Clinical assessment was performed with a Harris hip score. A reference group of 9 patients, who had undergone THA because of arthrosis, was chosen. In the fracture patients, we found a 9% decrease in bone mass and muscle volume of the middle femur. The BMD of the distal femur and proximal tibia showed a more marked osteopenia. There was no difference in these parameters between the two groups. In the reference group of operated arthrosis patients, we did not find any differences between sides postoperatively. After the operation, the fracture patients had a lower Harris score than the arthrosis patients, and this was most pronounced among those who had undergone osteosynthesis. The finding of a marked osteopenia after a femoral neck fracture, irrespective of treatment, but no bone loss after THA because of arthrosis, implies that patients with a femoral neck fracture are more sensitive to osteopenia, and that the bone loss is not proportional to the operative trauma. PMID- 9352044 TI - Different healing rates of bone autografts, syngeneic grafts, and allografts in an experimental rat model. AB - Matching of donors and recipients for tissue antigens is vitally important for successful transplantation of essentially all organs and tissues, the major exception being bone. The importance of tissue-typing for the healing of bone allografts remains, however, a controversial issue as development of both humoral and cell-mediated immunity against the grafted bone has been observed in some experimental systems. In the present study, we compared the healing patterns of frozen antigen-mismatched allografts, frozen antigen-matched allografts (syngeneic grafts), and fresh cortical bone autografts in an experimental rat model. Histomorphometry of the graft-host interface revealed that new bone formation started significantly earlier in autografts than in allografts or syngeneic grafts. By 2 weeks, the level of new bone formation in the syngeneic grafts had reached that in autografts. Antigen-mismatched allografts, however, continued to exhibit a retarded formation of new bone throughout the union process. These histomorphometric observations were confirmed by molecular biologic analyses for the mRNA levels of type I collagen, which increased earlier and reached a higher level in autografts than in allografts. Use of syngeneic grafts resulted in a longer persistence of type I collagen mRNA expression in the healing tissue than in antigen-mismatched allografts. No apparent differences were seen between allografts and autografts in the expression of type III collagen. No cartilage-specific type II collagen mRNA was observed, indicating that antigen-mismatching or preservation by freezing did not alter the basic mechanism of the interface healing process, although it did slow down the beginning of the process. The experiments suggest that a major antigen mismatch between donor and recipient affects the temporal gene expression of extracellular bone matrix and delays new bone formation at the graft-host interface of cortical bone allografts. PMID- 9352045 TI - Stabilization of osteochondral fractures: an experimental study comparing polyglycollic acid degradable pin with K-wire stabilization in rabbits. AB - Conventional metal implants may be unsuitable for the stabilization of osteochondral fractures as they may interfere with joint function and eventually require implant removal. We therefore compared the use of biodegradable implants with conventional metal ones in an animal experimental study conducted in skeletally mature rabbits. Biodegradable polyglycollic acids pins (PGA) 1.5 mm in diameter were used to stabilize an osteochondral fragment surgically created in the distal femur of rabbits. In another group of 36 animals, conventional metal K wire of the same diameter was used for stabilization. The animals were killed at intervals of 3 to 24 weeks. Satisfactory union of the fragments was noted in 92% of the PGA implants as compared with 50% with the metal implants group. No implant migration was seen in the PGA group, while migration was noted in all animals with the metal implants. Histological studies showed that in 80% of the cases fixed with PGA implants, the fragment was viable. In the metal group 33% of the fragments underwent fragmentation and necrosis. PMID- 9352047 TI - Rehabilitation of patients after hip disarticulation. AB - We review rehabilitation of patients after hip disarticulation operated on over the past 5 years. Sixty-two patients underwent 63 hip disarticulations: 24 had malignancies, 23 arteriosclerosis, 11 Buerger's disease, 3 diabetes and 1 uncontrollable infections. The mean age of tumour patients was 39 years (range 6 69 years), that of the vascular patients was 55 years (range 33-78 years). The postoperative mortality rate of the vascular patients was 16/37, and 0/24 of those with malignancies. There was one bilaterally operated patient with bilateral purulent coxitis from decubitus ulcers, who died on the 32nd post operative day. Complicated wound healing was observed in 5 of the 24 tumour patients, and in 24 of the 37 vascular patients. The strategy of prostheses fitting has a two-stage concept: temporary prostheses in the 1st or 2nd month, permanent prostheses in the 6th month. All 24 tumour patients were fitted and could walk, while only 2 of 37 vascular patients were able to walk with prostheses. In our experience the outcome is significantly dependent upon the primary illness: in vascular cases it is poor, while in malignancies it is fairly good. PMID- 9352046 TI - Bacterial colonization of bone allografts related to increased interval between death and procurement: an experimental study in rats. AB - Whereas organs from donors must be removed almost immediately after death to maximize organ viability in the recipient, there is a slightly longer window for tissue allograft recovery. To determine the maximum safe interval after death within which bone allografts may be harvested for clinical use, an experimental model was devised using adult Sprague-Dawley (SD) rats and duplicating cadaveric storage techniques. Allografts were procured at increasing time intervals after death. The grafts were then transplanted to 80 living SD rats, and the animals killed at 7 weeks to evaluate any increase in the risk of infection and bacterial colonization. None of the allografts procured within 48 h after death were colonized with bacteria, while 12% of grafts procured at 96 h and 50% of allografts procured at 1 week were colonized. The results suggest that it may be possible to extend the safe period within which cadaveric tissue may be procured for transplantation to up to 96 h following death, provided scrupulous measures to prevent and detect microorganism contamination are followed. PMID- 9352048 TI - Familial occurrence of hyperplastic callus in osteogenesis imperfecta. AB - There is a hypothesis that hyperplastic callus (HC) in osteogenesis imperfecta (OI) is not merely a rare complication but could actually be inherited, although this idea has not yet been investigated. We described two cases, a mother and son, with mild OI, normal scleral colour and no dentinogenesis imperfecta, who repeatedly had HC in their femur. Familial occurrence of HC was found in 13 cases in 5 families among 21 cases in 7 families with a familial background of OI in the literature (including this report). This is higher than the reported incidence of HC, 1.5% (5 cases of 333), and the mode of transmission is concomitant with autosomal dominant inheritance in all these families. Since a review of 47 cases in the literature shows that HC occurs independently of scleral colour and the degree of bone fragility, it may be an additional criterion for subdivision within each type of the Sillence classification. PMID- 9352049 TI - Flexor digitorum profundus avulsion with associated fracture of the distal phalanx. AB - Two unusual cases of flexor digitorum profundus avulsion with an associated fracture of the distal phalanx are reported. In one case the injury consisted of avulsion of a large bone fragment from the distal phalanx and an associated profundus avulsion from the fragment. The other case showed simultaneous shaft fracture of the distal phalanx with avulsion of the profundus tendon, which could not be included in the existing classification of profundus injuries. The latter injury was misdiagnosed initially. PMID- 9352050 TI - Attritional flexor tendon ruptures after a malunited intra-articular fracture of the distal radius. AB - Rupture of the flexor tendon following Colles fracture is uncommon. In all reported cases it occurred as a complication of an extra-articular, displaced fracture of the distal radius. We report a case in which flexor tendon rupture occurred 30 years after a comminuted intra-articular fracture of the distal end of the radius. There have been no reports of delayed flexor tendon rupture after an intra-articular fracture of the distal radius in young adults. PMID- 9352052 TI - Stress fracture of the tibia after total knee arthroplasty. AB - We present a case of total knee arthroplasty in which the patient suffered a transverse stress fracture of the tibia 2 weeks after replacement without significant injury. Plain radiographs were obtained on event presentation. Later radiographic examination showed the fracture and a periosteal reaction. The fracture healed after immobilising the limb in a cast for 2 months. This case illustrates well an insufficiency fracture to the occurrence of stress risers in a weakened bone when subjected to repetitive loading as a complication following total knee arthroplasty. PMID- 9352051 TI - Gorham massive osteolysis. AB - Gorham syndrome (massive osteolysis) is a very rare tumour-like lesion characterized by progressive osteolysis. The diagnosis must be confirmed by the microscopic finding of intramedullary angioma-like vascular structures. We report a case of a 15-year-old boy with a pathological fracture in his left humerus. Imaging modalities such as magnetic resonance imaging, computed tomography, angiography and bone scintigraphy failed to disclose to tumorous lesion that filled a cavity in the left humerus. After observing the boy's progress for 6 months, a temporary diagnosis of Gorham syndrome was made, and surgical treatment was chosen. After resection of the left humeral head and the proximal one-quarter of the humerus, thorough curettage was performed in the distal humerus and an intramedullary artificial humeral head fixed with adequate success. Pathological examination of the specimen revealed intramedullary haemangioma of the humerus. PMID- 9352053 TI - Unusual causes of scapular clicking. Lymphangioma of the thoracic wall and aneurysmal bone cyst of the scapula. AB - Scapular clicking with shoulder motion has been described for a variety of conditions. Two unusual cases, a lymphangioma of the thoracic wall in a 42-year old man and an aneurysmal bone cyst of the scapula in an 8-year-old boy, are presented. The lymphangioma was treated by marginal excision of the lesion and the aneurysmal bone cyst, by excision of the infraspinal portion of the scapula with resolution of symptoms. PMID- 9352054 TI - Restoration of elbow flexion 16 years after brachial plexus root avulsion. AB - To restore elbow flexion in brachial plexus lesions or cervical root avulsions, surgical nerve reconstruction can be attempted after approximately 6 months. If the reconstruction is not successful or was not performed, tendon transfer may improve the function of a paralysed limb. The selection of the muscle for transfer can be influenced by strengthening potentially available muscles by a myo-feedback method. PMID- 9352055 TI - Production of nitric oxide (NO) during the oxidation of human oxyhemoglobin by nitrite: application of a NO-selective electrode for the measurement of NO. AB - By using a nitric oxide (NO) selective electrode system. NO produced during the oxidation of human hemoglobin by nitrite was monitored. When 160 microM oxyhemoglobin (in heme) was reacted with 500 microM nitrite. NO was generated quickly at the initial lag phase of the oxidation of oxyhemoglobin by nitrite and decreased gradually during the second burst phase of the reaction. While the oxidation of oxyhemoglobin by nitrite proceeded in a sigmoidal manner including the initial lag phase and second burst phase. The maximal amount of NO produced under this condition was estimated to be 48 microM. According to the increase of nitrite concentrations added, the amounts of NO produced at the initial phase increased, being in good accordance with the increased rate of the oxidation of oxyhemoglobin. These results strongly suggest the critical role of NO in the oxidation mechanism of oxyhemoglobin by nitrite. PMID- 9352056 TI - Bis[2-(4-carboxyphenoxy)carbonylethyl]phosphinic acid (BCCEP): a new reagent for hemoglobin modification. AB - The synthesis and hemoglobin cross-linking studies of a novel organic reagent, bis[2-(4-carboxyphenoxy)carbonylethyl]phosphinic acid (BCCEP; 2) has been reported. The reagent was synthesized in four steps from hydroxybenzoic acid. The tri-sodium salt of BCCEP was employed to cross-link oxyHb, and the product was purified by DEAE-cellulose chromatography. The purified material was analyzed by SDS-PAGE, IEF, and HPLC analyses, which clearly showed the formation of covalent, intramolecular cross-links. While SDS-PAGE analyses of individual bands pointed to the molecular weight range of 32 kDa, the HPLC analyses suggested that the cross-links had formed between beta 1-beta 2 subunits. The oxygen equilibrium measurements and the Hill plots were performed on the purified bands to assess oxygen affinity as well as cooperativity of oxygen binding of the modified hemoglobins. All bands corresponding to modified hemoglobins showed significantly reduced oxygen affinity as compared with that of cell-free hemoglobin, as desired. The modified hemoglobins, however, exhibited somewhat reduced oxygen binding cooperativity as contrasted with human stroma-free hemoglobin. Molecular dynamics simulation studies (Insight II/Discover/Biosym) on the Reagent-HbA0 complex suggested that the most likely amino acid residues involved in the cross linking are Lys82 or N-terminal Val1 on one of the beta chains, and Lys144 on the other. PMID- 9352057 TI - Validation of the heat treatment step used in the production of diaspirin crosslinked hemoglobin (DCLHb) for viral inactivation--effect of crosslinking. AB - Two experiments were performed to assess viral inactivation during the crosslinking and heat treatment steps of the DCLHb manufacturing process. Stroma free hemoglobin (SFHb) collected from a large scale manufacturing lot was tested in a 1:680 scaled down system in which the key parameters used in the manufacturing process were replicated. In the first study Porcine Parvovirus (PPV), a non-enveloped virus, was used to assess inactivation, while in the second study Bovine Viral Diarrhea Virus (BVDV), an enveloped virus, was utilized. In both experiments, the SFHb solution was deoxygenated and an aliquot of virus suspension was added. To initiate the crosslinking reaction, a solution of bis (3,5-dibromosalicyl) fumarate (DBBF) in HEPES buffer was added to the test solution. In both experiments the reaction times and the degree of crosslinking were normal. After crosslinking, the reaction mixtures were heated to 74 +/- 1 degrees C over 30 minutes, held at 74 +/- 1 degrees C for 90 minutes, and cooled to less than 10 degrees C over 30 minutes. In each experiment the degree of crosslinking of final product was 100% and yield of hemoglobin recovery was normal. Samples were removed prior to crosslinking, after crosslinking and before, during and after heat treatment for determination of virus titer and evaluation of key process parameters. The results from these experiments were consistent with those obtained from the full scale manufacturing process for the deoxygenation, crosslinking and the heat treatment step during the production of DCLHb. The results of virus assays showed that crosslinking has no effect on viruses and their subsequent inactivation by heat treatment. PMID- 9352058 TI - Molar masses and structure in solution of haemoglobin hyperpolymers--a common calibration of size exclusion chromatography of these artificial oxygen carriers. AB - We are developing artificial oxygen carriers for medical use, based on synthetic polymers--so-called hyperpolymers--obtained by cross-linking mammalian haemoglobins. One requirement with respect to the polymers is that they should not increase the oncotic pressure of blood remarkably--this can be realized by high molecular weights of the polymers with a narrow distribution. They may act as a oxygen transporting blood additive, and--in combination with a plasma expander--as a blood substitute. Another important and desired property of the artificial oxygen carrier is a low viscosity, which--first--is due to a high degree of uniformity of the polymer size (or molar mass) distribution and--second -is influenced by the so-called structure in solution of the haemoglobin hyperpolymers. In this paper former determinations of molar masses--with size exclusion chromatography (SEC)--and of the structure in solution--using viscometric measurements--of hyperpolymers of human haemoglobin, synthetized with glutaraldehyde and with bis(thioisocyanato) benzenesulfonic acid as cross linkers, were extended to hyperpolymers of bovine and pyridoxylated porcine haemoglobin, cross-linked with glycolaldehyde. These determinations were done by applying a new iterative procedure. Within a range of error all SEC calibration curves found were the same for all hyperpolymers investigated. So-called MARK HOUWINK or structure in solution diagrams (logarithm of intrinsic viscosity versus logarithm of molar mass) yield equal straight lines for all the haemoglobin polymers. The first derivatives of these lines are the MARK-HOUWINK exponents which has a mean value of 0.38. These results indicate that there exists a common SEC calibration line for all different polymer haemoglobins produced with comparable preparative procedures. This calibration line differ significantly from that of native globular proteins--haemoglobin hyperpolymers are less compact--so a calibration of SEC with globular proteins for the determination of molar masses of haemoglobin polymers is erroneous. Furthermore, the structure in solution of the hyperpolymers is clearly different from that of flexible, randomly coiled, linear artificial polymers: hyperpolymers are more compact. A possible explanation is that the hyperpolymers--according to a great number of functional amino groups of haemoglobin-contain many intra-polymeric cross-links, and thus are at least import branched polymers or even macromolecular networks of the constituting haemoglobin "monomers". PMID- 9352059 TI - Modified polyacrylamide microspheres as immunosorbent. AB - Extracorporeal immunoadsorption system has been used for the specific removal of immunologically active substances from the blood. In this study, an attempt is made to utilise polyacrylamide microspheres as a matrix for immunoadsorption. Phenyl alanine is coupled onto the polymer beads using glutaraldehyde. These modified beads exhibit a high binding affinity for gamma-globulin compared to bare beads. The surface modified ones showed selective adsorption of immunoglobulins of IgG class. The C3 adsorption pattern is not altered significantly upon modification. Modified beads are found to be less hemolytic after modification. However, more studies are to be conducted for the development of a hemoperfusion column based on this modified matrix. PMID- 9352060 TI - Plasma-modified nylon meshes as supports for cell culturing. AB - The polymeric surfaces of three commercially available nylon films with mesh openings of 5 microns, 10 microns and 20 microns were treated with anhydrous ammonia gaseous plasma. Cells cultured on the plasma-treated nylon films have higher proliferation rate and assume morphology distinct from those cultured on the unmodified films. Of the three plasma-modified membranes, the one with 5 microns mesh openings supported a largest population of cell growth. The plasma treated nylon meshes provided a stronger anchorage for the collagen matrices formed within the mesh openings. Application of this collagen/nylon meshes for cell culturing is demonstrated. PMID- 9352061 TI - Effects of perfluorocarbon emulsions on cultured human endothelial cells. AB - Perfluorocarbons (PFCs) and their emulsions (PFCEs) were used in organ preservation before transplantation, but not in organ perfusion. Our purpose was to achieve organ perfusion with a PFCE at room temperature or at 37 degrees C, i. e. with oxygenation, to prevent damages related to reoxygenation after hypoxia. Therefore, we first investigated the effect of such emulsions on endothelial cells, the first cells to be in contact with the emulsion. A stem emulsion was prepared from perfluorooctyl bromide (90% w/v), emulsified with egg yolk phospholipids (2% w/v) and stabilized with a mixed fluorocarbon-hydrocarbon "molecular dowel" (1.4% w/v) (droplets of ca 0.2 micron in diameter). This emulsion was found to be stable when diluted with cell culture media or organ preservation fluids. Endothelial cells from human umbilical vein (HUVECs) were cultured in multiwell plates in M199 medium (with growth factors, 10% foetal calf serum and 5% human serum). Confluent cells were incubated overnight with 51Cr, washed and overlayed with M199 (control) or the above PFCE diluted 2x or 4x with M199 (test). After incubation, the cytotoxicity of the PFCEs was estimated by measuring 51Cr release and observing cell morphology by electron and light microscopy. The percentages of released 51Cr were identical to those of the control cells for the 2x, 3x or 4x diluted PFCEs at 4, 25 or 37 degrees C. After return to the M199 medium, the cells grew and multiplied normally. We conclude that the diluted PFCEs were devoid of cytotoxicity. The 2x diluted PFCE was however partially taken up by the cells: by microscopy, we observed intracellular PFC droplets and by density gradient analysis we found a slight increase in cellular density. The diluted PFCEs were compared to classical organ preservation solutions : HUVECs were incubated with UW (University of Wisconsin) or EC (EuroCollins) solutions at +4 and 37 degrees C (3, 17 or 24 h of incubation). The solutions were observed to be toxic to the cells under these conditions, with cell mortality after return to the M199 medium. This cytotoxicity may be attributed to the high K+ concentration of UW and EC, since similar assays performed on HUVECs with Hank's solution adjusted to 100 mM K+ showed a similar % of 51Cr release. UW and EC are therefore not acceptable as dilution media for PFCEs. PMID- 9352062 TI - Haemoglobin-enhanced mitotic division in cultured protoplasts. AB - Protoplasts from cell suspensions of albino Petunia hybrida cv. Comanche were cultured for 9 days in nutrient medium containing Erythrogen, a purified bovine haemoglobin solution (supplied at 10% w/v) at 1:50-1:500 (v/v). In some assessments, the non-ionic surfactant Pluronic F-68 (Poloxamer 188), was also added to the culture medium at 0.01-1.0% (w/v). Erythrogen at 1:50 (v/v) increased the mean initial protoplast plating efficiency (IPE; 18.5 +/- 0.8%, n = 5 throughout) by 64% (P < 0.001) above that of controls (11.3 +/- 0.4%). Supplementation of medium with 1:50 (v:v) Erythrogen and 0.01% (w/v) Pluronic F 68, increased the mean IPE (24.4 +/- 1.4%) by 92% (P < 0.001) over control (12.7 +/- 1.1%). Similar results were obtained for mesophyll protoplasts of Passiflora suberosa, with 1:50 and 1:100 (v/v) Erythrogen increasing the mean IPEs to 87% and 93% respectively, over controls. This beneficial and synergistic effect of Erythrogen with Pluronic F-68, on mitotic division of cultured Petunia and Passiflora protoplasts, should also facilitate the culture of isolated protoplasts and cells of other, agronomically-important, species. PMID- 9352063 TI - Changes in cell biochemistry in response to culture of protoplasts with oxygenated perfluorocarbon. AB - Superoxide dismutase (superoxide oxidoreductase; EC 1.15.1.1; SOD) was measured in enzymatically isolated protoplasts of Salpiglossis sinuata following culture in aqueous nutrient medium overlaying oxygen-gassed perfluorodecalin (Flutec PP6; BNFL Fluorochemicals, UK). SOD was extracted from harvested, lysed protoplast derived cells after 1, 3, 7 and 14 days of culture and assayed spectrophotometrically. Protoplasts cultured with oxygenated PFC (+/- s.e.m, n = 5) showed significant increases in mean SOD activity to 4.2 +/- 0.1 U after 1 day (P < 0.05) and 9.3 +/- 0.7 U after 3 days (P < 0.01), with a fall in mean SOD after 7 days (5.1 +/- 0.9 U), similar to control. The decrease in SOD after 7 days correlated closely with a progressive fall in pO2 in the PFC phase over the same period. In contrast, control protoplasts (medium alone) or protoplasts cultured in medium overlaying non-oxygenated PFC showed no significant changes in mean SOD activity over the 14-day culture assessment period. PMID- 9352064 TI - Differentiation of substrate-binding sites in pyruvate kinase by selective photoaffinity labeling. AB - The four substrate-binding sites in porcine liver pyruvate kinase have been labeled with the photoaffinity reagent 8-azido-2'-O-dansyl-[alpha-32P]ATP (AD ATP) under different experimental conditions. In the dark, the native pyruvate kinase was reversibly and competitively inhibited by AD-ATP, with KI = 2.8 microM and KADP = 0.18 mM. Under UV-irradiation, the enzyme was covalently labeled in the presence of Mg2+ by AD-ATP and inactivated irreversibly. Measurement of this photoinactivation process in the presence of various concentrations of ADP gave KI = 4.0 microM and KADP = 0.2 mM. A linear plot of the relative specific activity of the partially photolabeled enzyme after gel-filtration vs. the number of label per pyruvate kinase molecule introduced in the presence of Mg2+ shows that each covalent label completely inactivates a tetrameric pyruvate kinase molecule. In the strict absence of Mg2+, up to three substrate-binding sites in each pyruvate kinase molecule can be labeled by AD-ATP without decreasing enzyme activity. Subsequent addition of Mg2+ enables AD-ATP to label the remaining site and inactivate the enzyme. These observations show that there are one catalytic and three non-catalytic substrate-binding sites in each pyruvate kinase molecule. A probable structural reason for possible functional differentiation of intrinsically identical substrate-binding sites in all tetrameric enzymes is suggested. PMID- 9352065 TI - Metabolic inhibitors as tools to delineate participation of distinct intracellular pathways in enhancement of lactose-induced dissociation of neutrophil and thymocyte aggregates formed by mediation of a plant lectin. AB - Signaling processes in the course of the formation of the lectin-mediated aggregates may partake in conveying enhanced stability to the cell clusters. To prove the validity of this reasoning in a model, we have studied the impact of addition of three metabolic inhibitors (N-ethylmaleimide, nordihydroguaiaretic acid, and trifluoperazine) on lactose-dependent dissociation of cell aggregates, formed in the presence of the galactoside-binding mistletoe lectin. Using both human neutrophils and rat thymocytes to avoid measurement of responses restricted to a single cell type, an enhanced dissociation of lectin-formed cell aggregates was observed, when lactose and an inhibitor were present. Among the tested inhibitors, nordihydroguaiaretic acid and N-ethylmaleimide were more potent enhancers of cell dissociation than trifluoperazine. These results suggest that biosignalling pathways connected with lipoxygenase activity as well as the level of intracellular sulfhydryl groups confer further stability to lectin-dependent cell aggregates. The systematic evaluation of inhibitors for defined activities is thus suggested as a tool to disclose the nature and the contribution of individual signaling mechanisms to post-binding effects following lectin initiated cell contact formation. PMID- 9352066 TI - Design of novel analogue peptides with potent fungicidal but low hemolytic activity based on the cecropin A-melittin hybrid structure. AB - In order to design synthetic peptides with potent antifungal activity but low cytotoxic activity under physiological conditions, several analogues of the previously reported cecropin A (CA)-melittin (ME) hybrid peptide, CA(1-8)-ME(1 12), were synthesized. These analogues were designed by analysis of the alpha helical wheel diagram of CA(1-8)-ME(1-12). Antifungal activities were measured by growth inhibition of the yeast Trichosporon beigelii and by hemolytic assay with human red blood cells, respectively. Substitution of Thr for Lys at position 18 and 19 of CA(1-8)-ME(1-12) caused a dramatic reduction in hemolytic activity. Two analogue peptides (analogue I and III) showed more potent antifungal and lower hemolytic activity than the original peptide. To study the antifungal mechanism of these peptides, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed with the most powerful antifungal analogue I peptide designed in the present study. As determined by propidium iodide staining, fungal cells treated with analogue I or melittin showed higher fluorescence intensity than those treated with the weak antifungal peptide, cecropin A. By confocal microscopy the analogue I was detected in the intracellular region as well as the in cell membrane. These facts suggested that the antifungal function of this novel peptide analogue acts by pore formation in the cell membrane. PMID- 9352067 TI - Effects of hydrostatic pressure on the activity of rat ribosome and cell-free translation system. AB - The effects of high hydrostatic pressure on the protein synthesis activity of rat liver ribosome and the reconstructed cell-free translation system were studied. The results indicated that the activity of the ribosome decreased with the increase in applied pressure and the activity was totally lost as pressure went up to 2,400 bar, but there was a plateau approximately from 300 to 1,200 bar where the activity only had a minor change. The activity of the cell-free translation system seems to be more sensitive to pressure. Its activity was entirely lost when the pressure was up to 900 bar. The activity of rat ribosome and the cell-free translation system treated by pressure below 900 bar could be almost completely restored by incubation after releasing the pressure, but could be partially restored at higher pressure. PMID- 9352068 TI - Competitive inhibition of mouse brain nitric oxide synthase by amiloride: a case for enzyme cross-inhibition. AB - Amiloride, a diuretic and antihypertensive drug, inhibits Na+ transporting systems, diamine oxidases and the human urinary plasminogen activator. Present results indicate that amiloride is a competitive inhibitor of mouse brain nitric oxide synthase (NOS; Ki = 4.5 x 10(-4) M, at pH 7.5 and 37.0 degrees C), but not a nitric oxide (NO) precursor, representing a case for enzyme cross-inhibition. The decreased levels of NO, as a consequence of NOS inhibition, might induce some clinically-adverse amiloride reactions, such as an unexpected reduced antihypertensive effect. PMID- 9352069 TI - Single-strand breaks and repair in nuclear DNA in the presence of hydralazine assayed by the nucleoid technique. AB - The nucleoid sedimentation assay was used to study hydralazine-induced DNA structural changes and repair in the fibroblasts cultured in vitro. The drug induced a dose dependent loss in negative DNA supercoiling due to the physical breakage of the DNA. Relaxation of supercoiled DNA resulted in the nucleoids sedimenting with lower velocities than those of undamaged control cultures. Repair incubation of the cells did not cause the restoration of DNA supercoiling to control level. Unsuccessful repair of DNA damaged by hydralazine may result in maintaining the damaged DNA in the cell which could have immunologic consequences. PMID- 9352070 TI - Chaperone-like function of lipocortin 1. AB - Lipocortin 1 (LC1) is a 37 kDa member of the annexin family of proteins. It has been proposed to act as a mediator of some of the actions of glucocorticoids in anti-inflammatory and immune suppressive functions. LC1 has been shown to play a role in cell proliferation, apoptosis, and differentiation. However, the exact biological functions of LC1 still remain obscure. Here it is shown that LC1 displays a chaperone-like function. Stoichiometric amounts of LC1 suppressed thermally induced inactivation and aggregation of the test enzymes citrate synthase and glutamate dehydrogenase. LC1 was also effective in refolding guanine hydrochloride-denatured glutamate dehydrogenase, as judged by circular dichroism spectroscopy. PMID- 9352071 TI - Integrin expression and collagenase activity of RSV-transformed Syrian hamster fibroblasts, differing in spontaneous metastasizing. AB - The expression of extracellular matrix (ECM) specific receptors, integrins, and the activities of type IV collagenase and interstitial collagenase were investigated in two strains of oncogenically transformed fibroblasts, drastically differing in spontaneous metastasizing. Both strains were shown to express quite limited patterns of integrins. Of those, alpha 5 beta 1 integrin is greatly reduced on highly metastatic (HM) cells, while the expression of alpha v beta 3 is strongly suppressed on lowly metastatic (LM) fibroblasts. No differences between the strains were found in either intracellular or secreted activities of type IV collagenase, while the activity of interstittial collagenase, secreted by HM cells, was twice as much as secreted by LM cells. The results imply the role of cooperated modifications of integrin-directed cell-ECM interaction and collagenase activity in establishing a metastatic phenotype. PMID- 9352072 TI - Formation and characteristics of an unusual lambda-DNA species. AB - An unusual DNA species, termed as DNA species A, has been isolated and purified from thermal-denatured lambda-DNA Hind III by Sephadex G-200 gel filtration. Our studies indicate that DNA species A is resistant to DNase I digestion and has a higher melting point. The new DNA species showed a lower absorbency at 260 nm, and a lower fluorescence quantum yield after interaction with ethidium bromide (EB) than native double-stranded lambda-DNA. CD spectrum of DNA species A consists of a broad positive band centered at 245 nm and a weak negative band at 220 nm. transmission electron microscope (TEM) visualizations showed that their lengths of DNA species A fell mainly in three regions (300-500 nm, 750-1000 nm and 1500 nm) that corresponded to three fluorescence bands in the EB-stained gels. Their apparent width and height were 65-75 nm and 2.2 nm respectively as observed by images of atomic force microscope (AFM). PMID- 9352073 TI - Immunohistochemical analysis of heme oxygenase-I in rat liver after ischemia. AB - Total hepatic ischemia was induced by clamping the hepatic artery, portal vein, and bile duct. After 15 min hepatic ischemia, we examined a time course of the changes in the steady-state levels of heme oxygenase-1 (HO-1) mRNA and protein in the livers. The levels of HO-1 mRNA was transiently induced and peaked at 4 h after the start of reperfusion. In addition, a pattern of the time course of HO-1 protein levels was similar to that of HO-1 mRNA levels. Immunohistochemical analysis clearly showed that the localization of HO-1 protein induced by hepatic ischemia was restricted to the hepatocytes in the pericentral vein. PMID- 9352074 TI - Characterization of a beta-lactamase from Mycobacterium smegmatis SN2. AB - Beta-lactamases have been reported to be largely responsible for beta-lactam resistance in Mycobacteria. We report the characterization of a cell-associated beta-lactamase from Mycobacterium smegmatis. The enzyme hydrolyzed the "beta lactamase-stable" oximinocephalosporins. Nitrocefin was the best substrate. 6 Beta-iodopenicillanate, clavulanate and sulbactam were effective inhibitors, whereas the Ki value for aztreonam was high. From its substrate and inhibitor profile, the enzyme appeared to be a cephalosporinase of group 2e. PMID- 9352075 TI - Regulation of CREB phosphorylation by cAMP and Ca2+ in parotid acinar cells. AB - Since various secretory stimuli regulate not only secretion but also protein, RNA, and DNA syntheses in salivary glands, we evaluated the effect of secretory stimuli on the phosphorylation state of CREB (cAMP response element-binding protein). Isoproterenol, forskolin, and CPS-cAMP markedly stimulated the phosphorylation of CREB in parotid acinar cells, and PKA inhibitors H-8 and H-89 dose-dependently inhibited it. In contrast, carbachol (CCH) and A23187 decreased CREB phosphorylation, but CCH did not decrease it in the absence of extracellular Ca2+. Although protein phosphatase inhibitor calyculin A alone markedly increased the phosphorylation, it could not prevent CCH-induced dephosphorylation of CREB. CaM kinase IV, a putative protein kinase for CREB in response to Ca2+ elevation, was undetectable in parotid acinar cells. PMID- 9352076 TI - Dual temperature model for the estimation of energetics parameters for acetylcholinesterase inhibition by cyclophosphamide. AB - The present work addresses the 'dual temperature' model for the estimation of Gibb's free energy change (delta G), enthalpy change (delta H), heat of activation (delta H.) entropy change (delta S), temperature coefficient (Q 10) and activation energy (Ea) of chicken brain acetylcholinesterase (AChE) inhibited by cyclophosphamide monohydrate (CP). In this investigation, the PZ factor (number of sterically and energy wise favorable collisions occurring between CP and AChE) has also been studied. The inhibitor has considerably increased all energetics parameters except delta S and Q10. These results are in general agreement with the data from the other reported studies. The significance of the use of CP in cancer therapy and various aspects of thermodynamic parameters have also been discussed. PMID- 9352078 TI - Regulation of progesterone biosynthesis in the human placenta by estradiol 17 beta and progesterone. AB - Ex vivo addition of estradiol 17 beta to first trimester or term human placental minces caused a significant increase in the quantity of progesterone produced. Addition of an aromatase inhibitor, CGS 16949 A, or the estrogen receptor antagonist, ICI 182780, significantly inhibited progesterone production confirming the role of estradiol 17 beta in the regulation of progesterone synthesis in human placenta. RU 486 and ZK 98299, which are antagonists of progesterone receptor, significantly modulated progesterone synthesis in the human placenta but exhibited paradoxical effects on the first trimester and term placenta. We conclude that progesterone synthesis in the human placenta is regulated by estradiol 17 beta and progesterone. This is the first report providing evidence for autoregulation of progesterone synthesis in the human placenta. PMID- 9352077 TI - Isolation and characterization of anti morphine analgesic peptide from canine brain. AB - The anti-morphine peptide from canine brain, which is homologous to alpha subunit of hemoglobin of canine, has been isolated and characterized. The canine brain was homogenized and extracted in 0.1 M acetic acid. The supernatant was collected, and purified by chromatography of Sephadex G-50 and ion-exchange and RP-HPLC. A polypeptide was then obtained, which is termed as AMP5. It shows prominent antimorphine analgesic activity. AMP5 has an apparent molecular mass of 10.23kD. The isoelectric point was estimated to be around pH 6.7 by IEF. The amino acid composition of AMP5 and sequence of the N-terminal 50 amino acid residues was determined. PMID- 9352080 TI - Purification of recombinant human precursor acid alpha-glucosidase. AB - Large quantities of recombinant acid alpha-glucosidase are needed for in vivo experimentation of enzyme replacement therapy in Pompe disease. We describe a new purification method for the purification of this recombinant enzyme from tissue culture medium consisting of concanavalin A affinity chromatography, hydrophobic interaction chromatography, affinity chromatography on Superdex, and anion exchange chromatography. The new method is amenable to scale up, and has increased speed, and improved reproducibility with similar high yield and purification efficiency when compared to previous methods. PMID- 9352079 TI - Thermal stabilization of carboxypeptidase A as a function of pH and ionic milieu. AB - In this study we investigated the contribution of pH, phosphate anions and salt concentration to the catalytic and structural thermostability of the carboxypeptidase A (CPA). The concentration of 75-100 mM phosphate as well as neutral pH values were found to be optimal for stabilizing CPA at high temperatures. Although moderate concentrations of sodium chloride had no effect on thermal stability, high concentrations of the salt destabilized the enzyme. The experimental results and theoretical analysis suggested that the main contribution to heat stabilization of CPA is related to intramolecular electrostatic interactions and Arginine and/or Lysine are the putative groups able to bind phosphate and stabilize the enzyme molecule against thermal denaturation. PMID- 9352081 TI - Kinetic properties of cytosolic fructose 1,6-bisphosphatase from grapefruit. Effect of citrate. AB - cFBP is studied for its affinity to Mg++ and Fru-1,6-P2. The affinity for Mg++ is not very high with a Km of 0.24 +/- 0.01 mM. High concentrations of Mg++ are inhibitory. The saturation curve for Fru-1,6-P2 is hyperbolic with a Km of 0.54 +/- 0.014 microM. The presence of citrate (10 mM) induces a sigmoidal curve, modifying both Vmax and S0.5. Citrate affects the allosteric properties of cFBPase: at low substrate concentration cooperativity becomes negative while at higher concentration it is positive. Addition of higher concentrations of Mg++ shows a synergistic effect with citrate, decreasing of the affinity for Fru-1,6 P2: S0.5 equals 7.6 +/- 0.25 mM, 9.0 +/- 0.86 mM and 21.5 +/- 1.46 mM in presence of 5, 7.5 and 10 mM Mg++, respectively. PMID- 9352082 TI - Alteration of intracellular free calcium and acylphosphatase levels in differentiating SH-SY5Y neuroblastoma cells. AB - Levels of acylphosphatase isoenzymes and free intracellular calcium have been investigated in cultured SH-SY5Y human neuroblastoma cells under stimulation with all-trans retinoic acid and phorbol-12-myristate-13-acetate. Under these conditions morphological and functional characteristics demonstrated the differentiation of SH-SY5Y cells towards neuronal phenotype. Retinoic acid treatment caused a progressive and synchronous increase of the organ common-type acylphosphatase and of free intracellular calcium but not of the muscle-type acylphosphatase. Phorbol-12-myristate-13-acetate treatment gave rise to a peak of the muscle-type acylphosphatase levels during the early differentiation stage whereas organ common-type isoenzyme and free calcium levels show a pattern similar to that observed in retinoic acid-treated cells. These evidences indicate that the two acylphosphatase isoenzymes play different roles in SH-SY5Y differentiation and that during this process the expression of organ common-type acylphosphatase increases in a synchronous way with intracellular free calcium concentration. PMID- 9352083 TI - Binding characteristics of SLE anti-DNA autoantibodies to modified DNA analogues. AB - Native calf thymus DNA was modified with furocoumarins and reactive oxygen species (ROS). The modifications were probed by UV & Fluorescence spectroscopy, thermal denaturation and hydroxyapatite chromatography. In CD spectroscopy changes in the ellipticity of the DNA molecule were observed as a result of modification. The binding specificity of naturally occurring anti-DNA antibodies with modified DNA molecules was assessed by ELISA and quantitative precipitin titration. The affinity of anti-DNA antibodies for modified conformers was found to be quite high. PMID- 9352085 TI - Cloning of the human cDNA sequence encoding the NADH:ubiquinone oxidoreductase MLRQ subunit. AB - A cDNA clone encoding human NADH:ubiquinone oxidoreductase (complex I of mitochondrial respiratory chain) MLRQ subunit was isolated from human fetal liver cDNA library. The clone contained an open reading frame of 246 by which predicted a protein comprising 81 amino acids with a calculated molecular weight of 9,370 Da. The deduced amino acid sequence exhibited 95% homology (88% identity and 7% favored substitution) to that of bovine MLRQ subunit. Northern analysis revealed that the cDNA clone hybridized with a 0.7 kb mRNA species which was present in all tissues examined. The expression level of the 0.7 kb mRNA in heart, skeletal muscle, and brain was higher than in other organs. Human MLRQ cDNA could cross hybridize with the genomic DNAs from various species. PMID- 9352084 TI - Carbohydrate moiety of Plasmodium falciparum glycoproteins: the nature of the carbohydrate-peptide linkage in the MSP-2 glycoprotein. AB - Metabolic labelling of Plasmodium falciparum parasites with [3H]GlcN, [3H]Man, [3H]Gal and [3H]ethanolamine, and subsequent purification by SDS-PAGE of the labelled material provided effective labelling of the MSP-1, 195 kDa, and MSP-2, 42-53 kDa, glycoproteins. Reductive beta-elimination of the MSP-2 released from the gel consisted of glycopeptides containing labelled sugars. Processing of the eliminated components and identification of the sugar residues demonstrated the presence of N-acetylglucosaminitol and N-acetylgalactosaminitol amongst other labelled sugars. Reductive beta-elimination with sodium hydroxide-sodium borotritide-borohydride showed the presence of glucosaminitol and alanine in the hydrolysis products. The MSP-2 was retained on solid phase wheat-germ agglutinin and was released from the lectin by treatment with GlcNAc. Upon treatment with O glycanase the MSP-2 glycoprotein released labelled amino sugar, and derived oligosaccharides on treatment with exoglycosidases released labelled components corresponding to the metabolically incorporated sugars. Labelled Gal was incorporated into the MSP-2 glycoprotein using [3H]UDP-Gal and galactosyltransferase. The galactosylated glycoprotein released labelled Gal upon treatment with beta-galactosidase. The results of the present study suggest that the carbohydrate chains of the MSP-2 glycoprotein are attached to the protein backbone via GlcNAc- and GalNAc-serine/threonine in O-glycosyl linkage and the glycoprotein has terminal GlcNAc and Gal residues. The carbohydrate moieties of MSP-2, glycoprotein consist mainly of short chains linked to the protein core. PMID- 9352086 TI - Expression of chloroplastic genes during autumnal senescence in a deciduous tree Populus deltiodes. AB - In Populus deltoides, a deciduous tree, the development on new leaves starts in the month of March, the leaves reach maturity by October and fall by December. Changes in the composition and function of the photosynthetic apparatus were analysed during autumnal senescence. With the progress of senescence, there was an initial increase followed by a decrease in the steady state levels of psbA, psbD/C and psaA/B gene transcripts. Decrease in the steady state level of D1 protein was faster than that of Cytochrome f. The decline in LHCP level was seen only during late senescence. Although the leaves continue to look green and healthy till late November, the electron transport driven by individual photosystems started declining by October end suggesting the onset of senescence. PMID- 9352087 TI - Unusual AMP-deaminase solubilization from teleost fish white muscle. AB - A previous study described an unusual influence of neutral salts on the behavior of trout muscle AMP-deaminase (AMPD) in its interactions with subcellular particulate matter (Lushchak and Storey 1994, Fish Physiol. Biochem. 13: 356 368). The present study shows that this behavior is also shared by the muscle enzyme of two other fish species, sea scorpion (Scorpaena porcus) and corb (Sciena umbra), indicating that this describes a principle for AMPD interaction with cellular particulate material. AMPD binding to particulate matter increased with increasing KCl concentration through the physiological range (100-200 mM), but at higher salt concentrations the amount of bound enzyme was reduced. The pattern of binding was not influenced by hydrophobic interactions since addition of the nonionic detergents, Triton X-100 or Tween-80, did not alter the distribution of bound versus free enzyme although both detergents, at low concentrations, enhanced enzyme maximal activity. AMPD binding to particulate matter was also influenced by pH, the amount of free enzyme rising by nearly 3 fold as pH fell within the physiological range from 7.5 to 6.5. It is concluded that neither electrostatic nor hydrophobic forces alone can account for the unusual solubilization of AMPD from fish muscle and it is possible that the effect is also related to ion-induced conformational changes in the structure of AMPD and/or of the myosin to which the enzyme binds. PMID- 9352088 TI - Chlamydomonas U2, U4 and U6 snRNAs. An evolutionary conserved putative third interaction between U4 and U6 snRNAs which has a counterpart in the U4atac-U6atac snRNA duplex. AB - The spliceosomal UsnRNAs U2, U4 and U6 from the green alga Chlamydomonas reinhardtii (Cre) were sequenced using a combination of RNA and cDNA sequencing methods and were compared to other sequenced UsnRNAs. The lengths of Cre U6 and Cre U2 RNAs are similar to those of their higher plant equivalents. Cre U4 RNA is shorter (139 nt) than its counterpart from higher plants (150-154 nt), and contains stem IV and loop D which are absent, with the exception of the Tetrahymena U4 RNA, from the U4 RNAs of other unicellular organisms studied to date. Base-pairing interactions between U6 and U4 RNAs and between U6 and U2 RNAs, identical to those described for mammalian and yeast systems, are structurally feasible in the Cre system. In addition, based on comparative analyses of the predicted U4/U6 RNA duplex from various species, an evolutionary conserved third putative U6-U4 interaction was found. Interestingly, it can also be formed with the recently discovered U6atac and U4atac RNAs. This is a strong support in favor of the possible biological significance of this third putative interaction. Based on comparative analysis, an extension of the earlier described U6-U2 interaction patterns is also proposed. PMID- 9352089 TI - Acetylcholinesterase in Spirographis spallanzanii (Polychaeta: Sedentaria): presence of two dimeric membrane-bound forms. AB - In the annelid polychaete Spirographis spallanzanii two acetylcholinesterases, named DS and HSDS, were detected. They differ in relative amount, membrane anchoring and pharmacological properties. Studies with inhibitors evidenced complete inhibition of both acetylcholinesterases by 10(-3) M eserine and different sensitivities for edrophonium or procainamide. Both enzymes, sensitive to BW284c51, were unaffected by iso-OMPA; at variance, only the HSDS form underwent excess-substrate inhibition. DS and HSDS enzymes were solubilized by homogenization in a low-salt or high-salt-Triton X-100 buffer and then purified by affinity chromatography on edrophonium- or procainamide-Sepharose column respectively. According to gel-filtration chromatography, sedimentation analysis and SDS-PAGE, the least represented (30%) DS form is a G2 amphiphilic globular dimer (124-130 kDa, 6.0-7.0S) with S-S linked monomers (66 kDa). Phosphatidylinositol anchors give cell membrane insertion, self-aggregation and detergent (Triton X-100, Brij 97) interaction. The prevailing (70%) HSDS acetylcholinesterase is once again a G2 form similar to DS enzyme in its molecular size (117-125 kDa), sedimentation coefficient (6.0S) of the native form and presence of S-S linked subunits (66 kDa). However, it is likely attached to the cell membrane by involvement of strong electrostatic interactions. DS acetylcholinesterase displays moderate active site specificity with differently sized substrates. The HSDS form is inactive on butyrylthiocholine. DS and HSDS forms show a comparable catalytic efficiency (kcat/K(m)) approaching that of other invertebrate enzymes. The results suggest that DS and HSDS enzymes, likely encoded by distinct genes, are both functional in cholinergic synapses. PMID- 9352090 TI - GTP-dependent modification of a 21-kDa substrate with NAD+ in bovine brain soluble fraction is not ADP-ribosylation of small G-protein but tailing of tRNA. AB - Labeling of 21-kDa material was observed when bovine brain soluble fraction was incubated with [adenylate-32P]NAD+ in the presence of GTP. The 21-kDa substrate, slightly smaller than C3 substrate in size, was labeled even without C3 exoenzyme. GTP could be replaced by nucleoside triphosphates other than ATP while ATP inhibited the GTP-induced labeling of 21-kDa substrate. After incubation of the soluble fraction with [adenylate-32P]NAD+ in the presence of GTP, [32P]ADP and [32P]ATP were detected in addition to [32P]AMP and [32P]ADP-ribose while only the last two nucleotides were observed without GTP. The 21-kDa substrate was labeled with [alpha-32P]ATP even in the absence of GTP, suggesting adenylylation rather than ADP-ribosylation. The labeled 21-kDa substrate, was extractable by phenol, disappeared with RNase treatment but not with tryptic digestion. Alkaline treatment of the phenol extract yielded an equal mixture of 3'-[32P]CMP and 2' [32P]CMP. From these results we concluded that the 21-kDa labeling is a result of tRNA tailing with [alpha-32P]ATP generated from the [32P]AMP moiety of [adenylate 32P]NAD+. Results from reconstitution experiments using enzymes and tRNA purified from bovine brain soluble fraction, which are involved in this pathway, confirmed our conclusion. PMID- 9352091 TI - Synthesis of region-labelled proteins for NMR studies by in vitro translation of column-coupled mRNAs. AB - A method to synthesise region-labelled proteins for structural studies with NMR is suggested. The technique is based on in vitro translation of matrix-coupled mRNAs. Translation starts with unlabelled amino acids from the initiation codon of the mRNA and continues to the beginning of the region of interest. Here, the ribosomes pause while the tRNAs charged with unlabelled amino acids are replaced with tRNAs charged with isotope-labelled amino acids. Translation then proceeds through the region of interest until the ribosomes pause at its end. At this point aminoacyl-tRNAs are changed again. Translation is resumed with unlabelled amino acids and continues to the STOP codon of the mRNA, where the ribosomes pause. In the final step the complete, region-labelled protein is eluted from the column in almost pure form. The method is demonstrated for small scale synthesis of the DNA binding domain (DBD) of the glucocorticoid receptor (GR), where the DNA-recognising helix is labelled but the rest of DBD is unlabelled. The new technique can be generalised to allow a desired region in a protein to be isotope labelled. PMID- 9352092 TI - Copurification of dihydroxyacetone-phosphate acyl-transferase and other peroxisomal proteins from liver of fenofibrate-treated rats. AB - Dihydroxyacetone-phosphate acyl-transferase (DHAP-AT), a peroxisomal membrane bound enzyme that catalyzes the first step of ether-glycerolipid synthesis, was purified from liver of rats treated with fenofibrate, a peroxisome proliferator. The protocol first included isolation of peroxisomes, their purification through a discontinuous gradient and solubilization of membranes in CHAPS. DHAP-AT was further purified by four chromatographic steps, namely low-pressure size exclusion, cation-exchange, hydroxylapatite and chromatofocusing. The chromatofocusing step led to a 4000-fold increase in the specific activity of DHAP-AT with respect to the liver homogenate with a yield of about 0.2%. Trypsin digestion of a 64-kDa protein band upon SDS-PAGE resulted in a peptide sequence unknown in databases. A corresponding degenerated oligonucleotide was used as a probe in Northern blotting, and a transcript of 3.3 kb was detected in some rat tissues. Moreover, the overall procedure allowed co-purification of four major peroxisomal enzymes: urate-oxidase, catalase, multifunctional enzyme and palmitoyl-CoA oxidase, respectively. PMID- 9352093 TI - On the use of Zn2+ to discriminate endonucleases activated during apoptosis. AB - One approach to discriminate among specific DNases in apoptosis is to use inhibitors specific for each endonuclease. Zn2+ is known to inhibit Ca(2+)- and Mg(2+)-dependent endonuclease enzymatic activities during apoptosis. Acidic DNases were thought to be insensitive to Zn2+. In this paper, we analyse the effects of Zn2+ on activity of DNase II, either purified or in nuclei from lens fiber cells. These cells follow a physiological nuclear degeneration with DNase II accumulation in their nuclei. We show that Zn2+ is able to inhibit also this acidic endonuclease at a concentration of 1-6 mM. At a higher concentration of Zn2+, DNA is extensively degraded during the assay, masking the inhibition of the enzyme. This DNA degradation in the presence of Zn2+ has led to an overestimation of the activity of DNase II in studies of apoptosis. Hence, Zn2+ cannot be used to specifically identify one endonuclease among the different DNases involved in nuclear degradation during programmed cell death. PMID- 9352094 TI - Deletion screening and carrier detection in Duchenne muscular dystrophy in Polish population via direct analysis of DNA and RNA transcripts. AB - Analysis of 102 Polish Duchenne/Becker muscular dystrophy (D/BMD) patients was performed by 'multiplex' amplification of 22 fragments of the DMD/BMD gene and deletions were found in 55% of the patients. The data obtained using PCR were compared with results of 25 Southern blotting and hybridization experiments with cDNA probes and with immunostaining using anti-dystrophin antibodies. In order to determine more precise deletion breakpoints, additional experiments were performed on dystrophin transcripts isolated from peripheral blood lymphocytes. These data found direct application in carrier analysis in the respective families by detection or exclusion of aberrant cDNA fragments. Carrier detection was also performed by RFLP-PCR, analysis of polymorphic (CA)n repeats and single stranded conformational polymorphism (SSCP) for selected exons of the DMD gene. PMID- 9352095 TI - Metal-ion catalyzed oxidation affects fibrinogen activity on platelet aggregation and adhesion. AB - Exposure of fibrinogen to the Fe3+/ascorbate oxidative system resulted in structural modifications and altered functionality of the glycoprotein. The overnight treatment of fibrinogen by oxidants caused a 20-fold increase of carbonyl content with respect to the native protein. Formation of dityrosines as well as loss of tryptophan following fibrinogen oxidation were observed. The occurrence of conformational changes of the fibrinogen molecule as a consequence of the oxidative treatment was also established. Oxidized fibrinogen showed a distinct capability from the native molecule to mediate platelet aggregation and adhesion. The percentage of ADP-induced platelet aggregation decreased as a function of fibrinogen oxidative damage. Further, both unstimulated platelets and ADP-activated platelets showed a reduced ability to adhere to oxidized fibrinogen than to the native protein. These results suggest that oxidative treatment alters fibrinogen domains involved in the recognition and the binding of this molecule by the platelet receptor GP IIb/IIIa. PMID- 9352096 TI - Vanadates form insoluble complexes with histones. AB - Vanadium oxoanions are known to have a variety of physiological effects including insulin-like activity, inhibition of phosphotyrosine phosphatases, as well as direct interactions with a variety of cellular proteins, such as microtubules. In this study, vanadate was found to form insoluble complexes with histones, as well as other positively charged proteins, in a concentration dependent fashion. This interaction occurred over a 0.5-10 mM range which corresponds to the concentration range required for many of vanadate's known physiological effects. Results from precipitation experiments using vanadate solutions with or without the yellow-orange decavanadate indicated that the decamer form is primarily responsible for this precipitation. Vanadate was able to selectively precipitate histones from soluble chromatin as well as from a soluble bacterial protein extract to which a low concentration of histones had been added. Vanadate was also able to effectively precipitate histone from solutions as low as 0.006 mg/mL histone. Thus, the selective precipitation of histones and other positively charged proteins by vanadate can be utilized as a tool for protein purification. In addition, this interaction may provide insight into the mechanisms for the physiological effects of vanadate. PMID- 9352097 TI - Developmental expression of two rat sialyltransferases that modify the neural cell adhesion molecule, N-CAM. AB - Polysialylation of the neural cell adhesion molecule (N-CAM) reduces the efficacy of N-CAM-mediated homophilic binding and is regulated both during development and in regions undergoing neurogenesis or remodeling in the adult. Hamster PST-1 (PST) and rat STX are two related sialytransferases that catalyze the polysialylation of N-CAM. We have isolated a cDNA clone for the rat homologue of PST and compared its amino acid and nucleotide sequence to that of rat STX. This analysis revealed regions of high sequence similarity corresponding to the enzymatic domains of the two molecules. Other regions of lower similarity were used to generate specific probes for in situ hybridization. The distribution of PST and STX mRNAs, polysialic acid, and N-CAM were analyzed at three developmental stages. PST and STX mRNAs were expressed abundantly throughout the nervous system at embryonic day 15 and postnatal day 4 and were coexpressed in most tissues examined. In the adult brain, STX expression was reduced relative to PST and expression of both mRNAs was restricted to subsets of cells in areas undergoing constant synaptic rearrangement including hippocampus and olfactory system. The results suggest that both PST and STX participate in the polysialylation of N-CAM in vivo and that their expression levels are dynamically controlled during development and regeneration. PMID- 9352098 TI - Physiology and pharmacology of native glycine receptors in developing rat auditory brainstem neurons. AB - Glycinergic neurotransmission is mediated via inhibitory glycine receptors (GlyRs) which are heterogeneous during development. Electrophysiological studies performed on recombinant GlyRs have identified different pharmacological properties and attributed them to differences in their subunit composition. Here, we report on age-related changes in the response properties of native GlyRs in the mammalian brain. Whole-cell patch-clamp recordings were obtained from neurons of the medial nucleus of the trapezoid body (MNTB), a major relay station in the mammalian auditory brainstem. Experiments were performed in acute medullary slices of rats between postnatal day (P) 1 and P15, a period during which synapse maturation occurs. Glycine-induced currents were present throughout the period under investigation and displayed age-related modifications in their amplitude, kinetic characteristics, and sensitivity to drugs. Current amplitudes and GlyR desensitization behavior increased with age. The alpha 1 subunit-specific GlyR antagonist cyanotriphenylborate (CTB) was barely effective in reducing glycine induced currents during the first few postnatal days, yet a significant increase of the inhibitory effect occurred after the first postnatal week. This finding indicates that alpha 1 subunit-containing GlyRs become expressed only postnatally in the MNTB. Picrotoxin, which most effectively blocks recombinant alpha 2 homooligomers, reduced glycine-induced currents in neonatal MNTB neurons, suggesting that alpha 2-homooligomers may form native GlyR isoforms. Our results show that the physiology and pharmacology of GlyRs in the auditory brainstem underlie age-related changes which are most probably produced through a replacement of "neonatal" alpha 2 subunits with "adult" alpha 1 subunits. PMID- 9352099 TI - In vitro differentiation of chick spinal cord neurons in the presence of Reissner's fibre, an ependymal brain secretion. AB - The subcommissural organ (SCO), which belongs to the circumventricular organs, is a specialized ependymal structure of the brain that secretes glycoproteins into the cerebrospinal fluid (CSF) which condense to form a thread-like structure, the Reissner's fibre (RF). Regarding the presence of this ependymal brain secretion all along the central canal of the developing spinal cord, we analysed a putative developmental activity of RF on neuronal spinal cord cells. The effects of RF proper and soluble RF-material were examined in primary cultures of dissociated spinal cord cells from day 6 chicken embryos. In serum-containing mixed glial/neuronal cell cultures, both RF and soluble RF-material promoted neuronal survival. This effect was blocked by addition of specific antibodies raised against bovine RF into the culture medium. In serum-free neuron-enriched cultures, no neuronal survival activity was observed; however, under these conditions RF proper induced neuronal aggregation and neuritic outgrowth of spinal cord cells. Interestingly, neurites extending from the aggregates appeared mainly unfasciculated. Our results suggest a direct modulation of cell-cell interactions by SCO/RF glycoproteins and an indirect survival effect on neurons. These data strengthen the hypothesis of the involvement of SCO/RF complex in the development of the central nervous system (CNS) and are discussed regarding molecular features of SCO-spondin, a novel glycoprotein recently identified in this complex. PMID- 9352100 TI - Altered cell proliferation in the spinal cord of mouse neural tube mutants curly tail and Pax3 splotch-delayed. AB - The mutant mouse strains splotch-delayed (Pax3Sp-d) and curly tail (ct) develop neural tube defects (NTDs) in the lumbosacral region of the neuraxis. Some research has focused on cell proliferation around the time of posterior neuropore closure in these mutants; however, there are little data on the effects of NTDs on cell birth at later stages of development. To investigate the role neural tube closure might play in cytogenesis of the spinal cord, the thymidine analog 5 bromo-2'-deoxyuridine (BrdU) was injected into pregnant splotch-delayed and curly tail mice at various stages of gestation. The mean number of labelled cells in the dorsal and ventral halves of spina bifida and control embryos was then calculated per section and per mm2. Mutagenically separated PCR (MS-PCR), was used to ascertain the genotype of splotch-delayed embryos. Our data indicate that the peak proliferation dates, for both the dorsal and ventral regions of the cord, are similar in spina bifida and control embryos. However, the quantity of proliferation is significantly different between affected and unaffected embryos. In general, there are markedly fewer cells born in spina bifida embryos in early neural tube development, followed by a short period of equal proliferation, and culminating in a significant increase in cell proliferation later in gestation. This increase in proliferation results in a greater number of cells being born in spina bifida embryos compared to controls. Several possible explanations for this phenomenon are considered, including the hypothesis that the roof plate, or other factors induced by neural tube closure, might have an anti-mitotic activity. PMID- 9352101 TI - Evidence for increased seizure susceptibility in rats exposed to cocaine in utero. AB - Clinical observations indicate that cocaine use during pregnancy is a major health concern in the United States and may result in seizure-like behavior in the offspring. In the present study, we investigated whether prenatal cocaine exposure altered seizure thresholds measured in Sprague-Dawley rats, 60-90 days postnatal. In vitro postnatal studies, focusing on hippocampal tissue, revealed a reduced threshold for both electrical stimulation- and potassium-induced epileptiform discharges in slices from cocaine-exposed animals. Modest elevation of extracellular potassium concentration from 3 to 6 mM KCl elicited spontaneous epileptiform discharges in the majority of slices from cocaine-exposed animals (13/20) but rarely in slices from saline-exposed animals (2/18). In vivo studies on awake, freely behaving adult rats indicated a significant reduction in thresholds for both flurothyl- and kainic acid-induced seizures in cocaine exposed animals. Video-EEG monitoring during administration of kainic acid revealed reduced latencies to first 'electrographic seizure' and first 'electrographic seizure with behavior' in rats exposed to cocaine in utero compared to saline-treated controls. These studies provide strong experimental evidence that adult animals exposed to cocaine during gestation are at high risk for the development of seizure activity. PMID- 9352102 TI - Developmental expression of parvalbumin by rat lower cervical spinal cord neurones and the effect of early lesions to the motor cortex. AB - Expression of calcium binding proteins (CaBPs), increasing neuronal activity and phases of synapse elimination are widely believed to be linked during development. We have employed immunocytochemistry to study the expression of the CaBP parvalbumin (PV) during the postnatal development of the lower cervical spinal cord and investigated how early lesions to the motor cortex, at the onset of corticospinal synaptogenesis, perturb the normal pattern of PV expression. This study confirms previous observations that in normal rats PV-like immunoreactivity is confined to large sensory afferents for at least 10 days postnatally (P10) and that the adult pattern of expression emerges from about P18 and involves mainly dorsal horn neurones. However, the study has also demonstrated a transient wave of expression in ventral horn neurones which reaches a maximum between P14-18 and declines thereafter. Unilateral lesions made at P7 to the forelimb motor cortex, which sends an almost completely crossed projection to the spinal cord, resulted in reduced neuronal expression of PV in the lower cervical spinal cord contralaterally at a range of ages (P14-31). The median ratio of PV positive neurones contralateral/ipsilateral to the lesion in spinal cord segments C7 and C8 was significantly lower (p < 0.01) at 56.0% (34.5 76.8 95% confidence limits, n = 14) than in sham operated controls (99.7%, range 93.7-113.6, n = 5). The lesion affected the transient wave of expression seen in ventral horn neurones during the third postnatal week as well as dorsal horn expression at older ages. We conclude that there is considerable plasticity in PV immunoreactivity during spinal cord development. PV is transiently expressed by ventral horn neurones at an age when movement control is functionally maturing. Early cortical lesions disrupt this transient phase of expression but also alter mature patterns of PV localisation. This suggests a critical role for corticospinal pathways in guiding maturation of segmental spinal cord circuitry. PMID- 9352104 TI - Failure of neural tube closure in the loop-tail (Lp) mutant mouse: analysis of the embryonic mechanism. AB - Loop-tail (Lp) is unique among mouse mutants in failing to initiate neural tube closure at the cervical/hindbrain boundary (so-called 'Closure 1'), at the 5-7 somite stage. Lp/Lp embryos go on to develop a malformation that closely resembles cranio-rachischisis, the most severe neural tube defect found in humans. We investigated several possible embryological mechanisms that may underlie this failure of neural tube closure in Lp. The genotypes of Lp/Lp, Lp/+ and +/+ embryos from mixed litters were identified using the polymerase chain reaction to amplify a polymorphic microsatellite sequence that is very closely linked to Lp. At post-neurulation stages of development, Lp/Lp embryos have a shortened body axis, which could suggest a defect of axial elongation as the primary anomaly in Lp. However, we found that axial elongation is normal in Lp homozygotes prior to the stage of defective Closure 1, indicating that the shortened body axis of later embryos is a secondary effect of the neurulation anomaly, or an independent effect of the Lp mutation. Some workers have reported cell proliferation rates to be abnormal in later stage Lp/Lp embryos. We observed variations in [3H]thymidine labelling index, and mitotic index, between embryonic tissues, and between embryos at different somite stages. However, Lp/Lp, Lp/+ and +/+ embryos had closely similar cell proliferation parameters, arguing against a mechanism based on faulty embryonic growth. Thirdly, we tested the hypothesis that the defect in loop-tail results from an inability of the neural folds to become apposed, specifically at the site of Closure 1. By tying a silk suture around the embryonic axis, at the future site of Closure 1, we were able to effect convergence of the neural folds at this site. Neural fold closure failed to progress along the body axis in sutured Lp/Lp embryos, however, in contrast to operated Lp/+ and +/+ embryos which exhibited normal progression of neural tube closure. The embryonic defect in loop-tail appears, therefore, to involve either a general inability of the spinal neural folds to become apposed along the spinal region, or a defect in the process of neural fold fusion. PMID- 9352103 TI - Development of vomeronasal receptor neuron subclasses and establishment of topographic projections to the accessory olfactory bulb. AB - Previous studies of the adult vomeronasal system have shown that vomeronasal receptor neurons in the middle layer (expressing Gi alpha 2) and deep layers (expressing Go alpha) of the sensory epithelium project to the anterior and posterior parts of the accessory olfactory bulb (AOB), respectively. In the present study, the development of the two populations of vomeronasal receptor neurons and their segregated projections were investigated in the opossum, Monodelphis domestica. Antibodies to G proteins Gi alpha 2 and Go alpha were used to identify the two subpopulations of receptor neurons. The Gi alpha 2 immunoreactive (ir) cells and Go alpha-ir cells appeared between postnatal day 0 (P0) and postnatal day 3 (P3) and both types of cells increased in number during later development. The differential localization of Gi alpha 2-ir cells in the middle layer and Go alpha-ir cells in the deep layer of the VNO could be seen as early as P3 and became more prominent at later stages. The AOB was clearly identified at P10, and at this stage segregated projections of Gi alpha 2-ir fibers to the anterior part and Go alpha-ir fibers to the posterior part of the AOB were seen. The segregation of the two types of fibers in the AOB resemble that in the adult after P21. These results suggest that Gi alpha 2-ir and Go alpha-ir subpopulations of receptor neurons in the VNO develop in parallel, and that segregation of the two populations of receptor neurons in the VNO and the topographic projection to the AOB are established at very early stages during development. PMID- 9352105 TI - Expression of cerebellar specific glutamate and GABAA receptor subunits in heterotopic cerebellar grafts. AB - In the cerebellum, up- and downregulation of specific GABAA and NMDA receptor subunits coincide with granule cell migration and differentiation. In this study, in situ hybridization techniques with GABAA and NMDA receptor subunit specific probes were employed to assess whether the molecular phenotype of heterotopically grafted cerebellar granule cells corresponds to that of normal cerebellum. The cerebellar anlage of rat fetuses was stereotactically grafted into the rostral striatum of adult rats. Eight weeks after transplantation, analysis demonstrated acquisition of an adult differentiation status reflected by abundant GABAA alpha 6 and NR2C mRNA expression in granule cells. Complete lack of NR2B transcripts, molecular markers of immature granule cells, argues against persistence of undifferentiated cells. The data suggest that intrinsic cell-autonomous factors largely determine the molecular commitment of granule cells and that a restricted specific environment is not necessary to promote granule cell differentiation. PMID- 9352106 TI - Developmental changes in the dendritic architecture of salt-sensitive neurons in the nucleus of the solitary tract. AB - Recent studies have provided evidence that brainstem gustatory neurons undergo substantial dendritic growth during a period of postnatal development that coincides with the maturation of their response to salts, suggesting a relationship (perhaps causal) between the physiology and morphology of developing salt-sensitive neurons. In an initial effort to explore this issue, we used extracellular and intracellular recording and intracellular labeling techniques to examine the structure and function of individual gustatory neurons in the rostral nucleus of the solitary tract (rNST) of young (postnatal day [P] 22-28) and adult rats. We found that P22-28 cells that responded to all three of the salts in our taste array had a greater dendritic length, a greater cell volume, and more dendritic branches than the cells that responded to one salt. As a group, taste-sensitive neurons in P22-28 animals had a higher maximum dendritic branch order and a trend toward more dendritic branch points than gustatory neurons in adult animals. The dendritic arbors of P22-28 taste neurons that responded to all three salts were larger (greater surface area and volume), more extensive in the rostrocaudal axis and exhibited a higher maximum branch order, more branch points and higher swelling density than adult cells that responded to all three salts. These results demonstrate that the morphology of salt-sensitive gustatory neurons in developing animals is closely related to the number of salts that evoke a response. The data also support the postulate that gustatory neurons in the rat brainstem undergo substantial dendritic remodeling between the fourth week of life and adulthood. Dendritic remodeling may play an important role in the maturation of the rNST response to NaCl. PMID- 9352107 TI - Modulation of neurite branching by protein phosphorylation in cultured rat hippocampal neurons. AB - The control of branching of axons and dendrites is poorly understood. It has been hypothesized that branching may be produced by changes in the cytoskeleton [F.J. Diez-Guerra, J. Avila, MAP2 phosphorylation parallels dendrite arborization in hippocampal neurones in culture, NeuroReport 4 (1993) 412-419; P. Friedrich, A. Aszodi, MAP2: a sensitive cross-linker and adjustable spacer in dendritic architecture, FEBS Lett. 295 (1991) 5-9]. The assembly and stability of microtubules, which are prominent cytoskeletal elements in both axons and dendrites, are regulated by microtubule-associated proteins, including tau (predominantly found in axons) and MAP2 (predominantly found in dendrites). The phosphorylation state of tau and MAP2 modulates their interactions with microtubules. In their low-phosphorylation states, tau and MAP2 bind to microtubules and increase microtubule assembly and/or stability. Increased phosphorylation decreases these effects. Diez-Guerra and Avila [F.J. Diez-Guerra, J. Avila, MAP2 phosphorylation parallels dendrite arborization in hippocampal neurones in culture, NeuroReport 4 (1993) 412-419] found that protein phosphorylation correlates with neurite branching in cultured rat hippocampal neurons, and hypothesized that increased protein phosphorylation stimulates neurite branching. To test this hypothesis, we cultured rat hippocampal neurons in the presence of specific modulators of serine-threonine protein kinases and phosphatases. Inhibitors of several protein kinases, which would be expected to decrease protein phosphorylation, reduced branching. KT5720, an inhibitor of cyclic AMP-dependent protein kinase, and KN62, an inhibitor of Ca(2+)-calmodulin dependent protein kinases, inhibited branching of both axons and dendrites. Calphostin C and chelerythrine, inhibitors of protein kinase C, inhibited branching of axons but not dendrites. Treatments that would be expected to increase protein phosphorylation, including inhibitors of protein phosphatases (okadaic acid, cyclosporin A and FK506) and stimulators of PKA (SP-cAMPS) or PKC (phorbol 12-myristate 13-acetate), increased dendrite branching. Only FK506 and phorbol 12-myristate 13-acetate stimulated axon branching. A subset of these agents was tested to confirm their effects on protein phosphorylation in this preparation. Okadaic acid, FK506 and SP-cAMPS all increased protein phosphorylation; KT5720 and KN62 decreased protein phosphorylation. On Western blots, the position of MAP2c extracted from cultures exposed to okadaic acid was slightly shifted toward higher molecular weight, suggesting greater phosphorylation, while the position of MAP2c from cultures exposed to KT5720 and KN62 was slightly shifted toward lower molecular weight, suggesting less phosphorylation. We conclude that protein phosphorylation modulates both dendrite branching and axon branching, but with differences in sensitivity to phosphorylation and/or dephosphorylation by specific kinases and phosphatases. PMID- 9352108 TI - Proton-induced cation current in embryonic rat spinal cord neurons changes ion dependency over time in vitro. AB - A rapid increase in proton concentration [H+]0 induces Na+ conductance in a variety of cell types. Here we report that H+ trigger a cation-selective channel whose ion dependency changes over time in culture. Whole-cell recordings of ventral spinal cord neurons dissociated at E15 and cultured for up to 14 days revealed that more than 80% had H(+)-induced inward current responses exhibiting a rapid decay phase. The current response was activated beginning about pH 6.8. Following decay, several minutes were required for complete recovery. More modest decreases in pH, which by themselves failed to activate this current, depressed those triggered by effective changes in pH. The currents recorded from cells in culture for less than 7 days could be abolished completely in the absence of Ca2+ and persisted in Na(+)-free and Ba(2+)-containing solutions. Ensemble analysis of current fluctuations recorded at the peak of the current allowed us to estimate a unitary channel conductance of 7.0 pS and a mean open time of 4.1 ms. In neurons cultured 2 weeks or more, protons induced an inward current response with similar kinetic properties, but with [Na+]0 dependency. Thus, proton-activated cation conductance in embryonic rat spinal cord neurons is self-limiting and involves brief openings of cation-selective channels whose ion dependency changes over time in culture. PMID- 9352110 TI - Neonatally wounded skin induces NGF-independent sensory neurite outgrowth in vitro. AB - An in vitro model was established to investigate factors underlying the sensory hyperinnervation of neonatal rat skin wounds that has been observed in vivo (Reynolds and Fitzgerald, J. Comp. Neurol. 358 (1995) 487-489). Explants of normal and wounded rat dorsal foot skin were co-cultured with explants of embryonic chick or newborn rat dorsal root ganglia for 24 h and the number of sensory neurites counted. Explants of skin surrounding a wound made at birth were taken 3 (P3) or 10 (P10) days later and compared with normal skin of the same age. In addition, explants were taken from adult skin wounded 3 and 10 days earlier. At P3, normal skin induced weak neurite outgrowth (mean 13.1 +/- 2.1 neurites per ganglion explant) but skin that had been wounded 3 days earlier, at birth, induced three times more neurite outgrowth (37.8 +/- 3.3). Ten days after wounding at birth, neurite outgrowth was still substantial (40.9 +/- 3.3) although at that age (P10), even normal skin stimulates substantial growth (37.4 +/- 2.9). Normal adult skin also stimulated neurite outgrowth (28.7 +/- 0.45) but this was not increased by wounding 3 or 10 days earlier, and this was enhanced 3 days but not 10 days after wounding. Anti-NGF (nerve growth factor) added to the culture medium blocked the constitutive neurite stimulating activity from normal P10 and adult skin but was ineffective in blocking the neurite stimulating activity produced by neonatal wounding. It is concluded that skin wounding at birth results in release of one or more sensory neurotrophic factors that stimulate rat and chick dorsal root ganglia neurite outgrowth for at least 10 days, but which do not include NGF. PMID- 9352111 TI - Inhibition of glutathione synthesis can enhance cycloheximide-induced protection of developing neurons against axotomy. AB - Developing neurons depend for survival on target-derived trophic substances. These are thought to block the expression of a genetic program of cell death. Nevertheless, it is known that less orderly events such as oxidative stress are involved in neuron death. In vivo, retinal ganglion cell death induced by axotomy can be reduced by antioxidants. In this study, we investigated the effects of inhibiting glutathione synthesis by means of buthionine sulfoximine to characterize the influence of endogenous glutathione-dependent antioxidant systems on ganglion cell death. Moreover, since protein synthesis inhibition by cycloheximide has been shown to enhance glutathione synthesis in vitro, we studied the effects on cell death of intraocular injections of buthionine sulfoximine, cycloheximide and combinations of the two inhibitors. Cycloheximide's protective action did not seem to involve an increase in glutathione synthesis. Surprisingly, buthionine sulfoximine injected before cycloheximide enhanced its protective effects, whereas it inhibited them when injected later. We interpret our results as an interaction between death promoting effects of glutathione depletion through an elevation of free radical concentrations and cycloheximide-sensitive effects of oxidative stress through the synthesis of both death-inhibiting and death-promoting proteins. PMID- 9352109 TI - Developmental expression of synaptophysin, synapsin I and syntaxin in the rat retina. AB - Expression of synaptophysin, synapsin I and syntaxin was studied immunocytochemically in the developing rat retina using indirect immunoperoxidase technique. In the inner plexiform layer (IPL), syntaxin immunoreactivity appeared at postnatal day 1 (P1) whereas synaptophysin and synapsin I staining were first observed at P2. In the outer plexiform layer (OPL), synaptophysin appeared at P4, while synapsin I and syntaxin appeared at P8. In the case of synaptophysin, a punctate pattern of staining was observed from the time of its appearance (P4) in the OPL and from P12 onwards in the IPL. Synapsin I and syntaxin immunoreactivity in the OPL were of a low intensity throughout the development and in the adult stage. These findings are discussed in relation to synaptogenesis in the rat retina. PMID- 9352112 TI - Increased beta adrenoceptor activation overcomes conditioned olfactory learning deficits induced by serotonin depletion. AB - It was hypothesized that 5-HT2 receptors in the olfactory bulb prime the bulbar response to a beta adrenoceptor mediated unconditioned stimulus (UCS) during odor preference learning in 1-week-old rat pups. The ability of 4 mg/kg of isoproterenol + stroking and 6 mg/kg of isoproterenol + no stroking to induce normal odor preference learning in pups depleted of bulbar 5-HT in the present study supports the hypothesis. The inverted-U curve relation between UCS strength and learning also appears to occur within the bulb. PMID- 9352113 TI - Change of zinc distribution in rat brain with increasing age. AB - Zinc (Zn) accumulation in the brain of rats of various ages was studied to look into the significance of Zn for the development and function of the brain. The Zn concentration of the cerebral hemisphere was relatively low in 1- to 11-day-old rats. The Zn concentration of the cerebellum gradually increased after birth and reached nearly a plateau at 11 days old. At 48 weeks old, the Zn concentrations of the cerebral cortex and hippocampus formation were approximately twice that of the cerebral hemisphere at the early stage after birth and significantly higher than that of the cerebellum. When 65ZnCl2 was injected into two groups of rats at 5 days and 48 weeks old for comparison, 65Zn distribution in the brain of the former group was higher than that of the latter. In the neonatal rats, the highest concentration of 65Zn was found in the cerebellum, followed by the hippocampus formation, a Zn-containing neuron-rich region. In the adult rats, the highest concentration of 65Zn was found in the CA3 and dentate gyrus of the hippocampus formation. At 48 weeks, 65Zn distribution in the cerebellum was relatively low and at about the same level as in the cerebral cortex. These results suggest that Zn is highly demanded by the cerebellum, which develops rapidly after birth. The increase in Zn concentration with increasing age may reflect the Zn requirement for functioning as an neuromodulator as well as for brain development. PMID- 9352114 TI - Expression of HGF and cMet in the developing and adult brain. AB - Hepatocyte growth factor (HGF) was recently recognized as a potential neurotrophic factor in the developing brain. We studied expression of HGF and its receptor using Northern blot analysis and in situ hybridization for mRNA and double immunofluorescent laser confocal microscopy. HGF and cMet messages were abundant in the hippocampus of both human and rat brains. In this region, both messages were localized in the neuronal layer. Segregation of HGF predominantly in the hippocampal CA3-4 and cMet in CA1 supports the hypothesis that HGF may mediate important neurotrophic functions in both developing and adult brains. PMID- 9352115 TI - Cerebellar granule cells elaborate neurites before mitosis. AB - Neuronal birth and neurite outgrowth have been regarded as discrete, sequential stages of development. However, we recently found that sympathetic neuroblasts often elaborate axons before mitosis, in culture [E. Wolf, I.B. Black, E. DiCicco Bloom, Mitotic neuroblasts determine neuritic patterning of progeny, J. Comp. Neurol. 367 (1996) 623-635] and in vivo [E. Wolf, I.B. Black, E. DiCicco-Bloom, Central and peripheral neuroblasts elaborate neurites prior to division in vivo and in vitro, Soc. Neurosci. Abstr. 21 (1995) 785; E. Wolf, I.B. Black, E. DiCicco-Bloom, Mitotic neuroblasts engage in axonal outgrowth and pathfinding in vivo, Soc. Neurosci. Abstr. 22 (1996) 525]. Here, we report that cerebellar granule cells often divide with heritable neurites in vitro. Therefore, mitotic CNS precursors, in addition to peripheral neuroblasts, simultaneously undergo proliferation and process formation. Potentially, neurites on dividing precursors may allow target fields to influence directly the course of neurogenesis. PMID- 9352116 TI - Developmental plasticity of selected spinocerebellar axons. Studies using the North American opossum, Didelphis virginiana. AB - When the thoracic spinal cord of the opossum is hemisected at postnatal day 5 or 8, but not at day 12 or later ages, spinocerebellar axons which originate from spinal border cells, the sacral/coccygeal ventrolateral nucleus, and Stilling's nucleus grow through the lesion and reach the cerebellum. The critical period for such growth is comparable to that reported previously for spinocerebellar axons originating within Clarke's nucleus and for axons of the fasciculus gracilis, but shorter than that for most descending spinal axons. It appears, therefore, that differences exist in the ability of ascending and descending axons to traverse a lesion of their spinal pathway during development. PMID- 9352117 TI - Heavy-metal toxicity in an insect cell line. Effects of cadmium chloride, mercuric chloride and methylmercuric chloride on cell viability and proliferation in Aedes albopictus cells. AB - We evaluated the toxicity of CdCl2, HgCl2, and MeHgCl on the C6/36 cell line of Aedes albopictus. This cell line proved to be a suitable tool for studying heavy metal toxicity in insect cells. Since data on heavy-metal toxicity in invertebrate cell cultures are almost nonexistent, our results are discussed in relation to in vivo invertebrate and in vitro vertebrate studies. Viability and proliferation were assessed by dye exclusion and DNA quantification, respectively. Viability tests were carried out with and without 5% fetal calf serum in the medium. The three metal species decreased viability to different extents (MeHgCl > HgCl2 > CdCl2), and fetal calf serum had a protective effect. In serum-deprived cultures, LD50 values were 140.20, 2.51, and 2.08 mumol/L for CdCl2, HgCl2, and MeHgCl, respectively. For cultures with fetal calf serum, LD50 values were 149.71, 12.01, and 5.47 mumol/L, respectively. The viability curve for CdCl2 under serum-free conditions suggests the induction of a cell defense system. The three metal species also inhibited cell proliferation (MeHgCl > CdCl2 > HgCl2). The IC50 values were 1.75, 18.36, and 0.96 mumol/L for CdCl2, HgCl2, and MeHgCl, respectively. In summary, low MeHgCl concentrations caused both cell death and inhibition of cell proliferation; HgCl2 primarily disrupted the plasma membrane, whereas CdCl2 primarily inhibited cell proliferation. PMID- 9352118 TI - Liver cell culture methods to measure DNA alterations produced by chemicals and radiation. AB - Liver culture systems, both primary cultures of hepatocytes and replicating cell lines, can be used in a variety of ways to study the DNA-damaging effects of chemicals and radiation. The present report describes some of the available methods and their interpretation. PMID- 9352119 TI - Antibodies to oxidative DNA damage: characterization of antibodies to 8 oxopurines. AB - The 8-oxo-7,8-dihydropurines (8-oxopurines) are important cellular premutagenic lesions produced in DNA by free radicals. Specific antibodies were prepared to detect these lesions. For antigens, 8-oxo-7,8-dihydroadenosine (8-oxoAdo) and 8 oxo-7,8-dihydroguanosine (8-oxoGuo) were synthesized from the bromonucleosides, and the immunogens were produced by conjugating these to either bovine serum albumin or rabbit serum albumin by the periodate method. Polyclonal antibodies specific for the haptens were elicited from rabbits immunized with the BSA conjugates. The antibodies to 8-oxoAdo (anti-8-oxoAdo) and 8-oxoGuo (anti-8 oxoGuo) precipitated the homologous antigens in an Ouchterlony gel diffusion assay and no cross-reactivity was observed toward the normal nucleosides or to the heterologous 8-oxopurine. Specificity was also examined by hapten inhibition of antibody reactivity with the homologous conjugates using ELISA. For anti-8 oxoAdo, the IC50 for 8-oxoAdo was 8 mumol/L and 8-bromoadenosine, guanosine, and inosine did not inhibit, even at concentrations of 1.25 mmol/L. Similarly, the IC50 for anti-8-oxoGuo for 8-oxoGuo was 0.1 mumol/L. 8-Methoxyguanosine also inhibited the reaction but was about 500-fold less effective than the eliciting hapten. Other nucleosides tested did not inhibit at concentrations up to 100 mumol/L. Both antibodies could easily detect the corresponding damage in x irradiated f1 DNA at a dose of 7.5 Gy and both antibodies recognized the corresponding lesion in duplex DNA; however, with anti-8-oxoGuo the signal was reduced about 50% compared to single-stranded DNA. In order to determine the exact amount of each lesion produced in irradiated DNA, and to standardize the ELISA signal, both products were measured after alkaline phosphatase digestion of x-irradiated calf thymus DNA using high-pressure liquid chromatography (HPLC) coupled to an electrochemical detector. Anti-8-oxoGuo could detect ten 8-oxoG residues and anti-8-oxoAdo could detect two 8-oxoA residues per 10,000 nucleotides. Thus, these antibodies should be useful for the detection and measurement of 8-oxopurines in cellular DNA. PMID- 9352120 TI - Ineffectiveness of the presence of H-ras/p53 combination of mutations in squamous cell carcinoma cells to induce a conversion of a nontumorigenic to a tumorigenic phenotype. AB - Human tumor cells have properties in vitro or in surrogate hosts that are distinct from those of normal cells, such as immortality, anchorage independence, and tumor formation in nude mice. However, different cells from individual tumors may exhibit some, but not all of these features. In previous years, human tumor cell lines derived from different tumor and tissue types have been studied to determine those molecular changes that are associated with the in vitro properties listed above and with tumorigenicity in nude mice. In the present study, seven cell lines derived from human tumors were characterized for p53 and ras mutations that may occur in SCC tumor phenotypes and for tumor formation in nude mice. This investigation was designed to examine whether co-occurrence of mutated ras and p53 lead to a malignant stage in the progression process. None of the seven cell lines contained mutations in the recognized "hot spots" of the p53 tumor suppressor gene, but four had a nonsense/splice mutation in codon 126 and a mutation in codon 12 of the H-ras gene. The remaining three cell lines had p53 mutations in intron 5, in codon 193, and a missense mutation in codon 126, respectively. Four of seven cell lines were nontumorigenic; two of these cell lines contained a nonsense p53-126 mutation and mutated ras; one had a missense mutation at codon 126 but no mutated ras; the the fourth had only a p53 mutation at codon 193. Two of the nontumorigenic cell lines were converted to tumorigenicity after treatment with methyl methanesulfonate or N-methyl-N'-nitro N-nitrosoguanidine with no apparent additional mutations in either gene. Our analysis revealed that there was a high frequency of genetic diversity and mutations in both p53 and H-ras. There was also a lack of a causal relationship in the presence of mutations in p53 and the cells' ability to exhibit a malignant potential in nude mice. PMID- 9352121 TI - Validity of a model of cultured myocardial cells for assessment of cardioplegia. AB - Myocardial protection is usually studied in vitro on perfused heart preparations, but never directly on cultured cardiomyocytes. We evaluated a model of cultured newborn rat cardiomyocytes to study both the cytotoxicity and the protective effect against chemical hypoxia of three cardioplegic solutions (St Thomas' I, Bretschneider, St Thomas' II) under normothermic (37 degrees C) and hypothermic (4 degrees C) conditions. Cytotoxicity was evaluated in 50% and 100% concentrations of the cardioplegic solutions with incubation times from 90 to 360 min. Myocardial protection was studied in 50% cardioplegic solution with metabolic inhibitors. Immediate and late viabilities, after 24 h of recovery in the medium, were evaluated by simultaneous staining with fluorescein diacetate and propidium iodide. At 37 degrees C, the 50% concentration of the three cardioplegic solutions did not modify cell viability. At 37 degrees C, with 360 min of incubation, the 100% concentration of the St Thomas' I and Bretschneider solutions diminished immediate viability (mean +/- SD; medium 87% +/- 2%; St Thomas' I 58% +/- 5%; Bretschneider 37% +/- 8%; St Thomas' II 89% +/- 3%) as well as late viability (medium 69% +/- 2%; St Thomas' I 32% +/- 3%; Bretschneider 24% +/- 7%; St Thomas' II 65% +/- 4%). At 4 degrees C, immediate and late viabilities were unaffected by cardioplegic solutions. At 37 degrees C, after 360 min incubation time, metabolic inhibitors diminished immediate viability to 29% +/- 1% and late viability to zero. None of the three cardioplegic solutions used at 50% concentration prevented this effect. At 4 degrees C, immediate viability was not significantly affected by metabolic inhibitors (73% +/- 10%), but the use of Bretschneider cardioplegic solution seemed to be detrimental (53% +/- 9%). On the other hand, recovery phase after pretreatment with metabolic inhibitors with or without cardioplegic solutions for 360 min significantly diminished late viability (medium 63% +/- 7%; metabolic inhibitors 17% +/- 8%; St Thomas' I 17% +/- 6%; Bretschneider 8% +/- 6%; St Thomas' II 15% +/- 3%) and again cardioplegia was inefficient. In conclusion, in this in vitro model for the study of cardioplegic solutions, only pure concentrations of the St Thomas' I and Bretschneider solutions under normothermic conditions were cytotoxic. The well known protective effects of hypothermia against ischemia and reperfusion injury were both reproduced. Therefore, and even though cardioplegia failed to have any protective effect, probably owing to a severe metabolic inhibition, this model may be useful for studying myocardial protection. PMID- 9352122 TI - Comparative cytotoxicity of 5-aminosalicylic acid (mesalazine) and related compounds in different cell lines. AB - Nonsteroidal anti-inflammatory drugs can cause serious side-effects such as tubulo-interstitial nephritis. Mesalazine (5-ASA, 5-aminosalicylic acid) is used for the treatment of colitis ulcerosa, Crohn disease, and other diseases; it has been found to induce necrosis of both proximal convoluted tubules and renal papillaries. The comparative cytotoxicity of 3-, 4-, and 5-aminosalicylic acid, acetylsalicylic acid (AcSA), and the parent compound salicylic acid (SA) was investigated for the free acids and for their sodium salts. The interaction with endogenous glutathione (GSH) was also investigated. Four established cell lines were used: MDCK, LLC-PK1, NRK as renal cells, and HepG2 as hepatic cells. The free acid compounds were less toxic than their corresponding salts. Acidic 5-ASA was the most toxic of the three isomers in MDCK and LLC-PK1 cells, while NRK and HepG2 were more susceptible to acidic 3-ASA. Addition of NaOH modified the relative toxicity of 3-ASA and 5-ASA. The LLC-PK1 and HepG2 cells were more sensitive to the test chemicals as their salts than were the NRK and MDCK cells. SA and 5-ASA decreased the GSH content in renal cells and increased it in HepG2. GSH depletion with L-buthionine-(S,R)-sulfoximine enhanced the toxicity only for SA in NRK and for 5-ASA and AcSA in HepG2. No correlation between endogenous GSH and the susceptibility of MDCK and LLC-PK1 to the test compounds was observed. The results suggest that no typical nephrotoxic effect occurred. No explanation could be found for the tubulo-interstitial nephritis caused by 5-ASA therapy. PMID- 9352123 TI - Establishment of a rat Sertoli cell line that displays the morphological and some of the functional characteristics of the native cell. AB - Primary rat Sertoli cells are widely used as a model for mechanistic and toxicological studies, since they are often the target of toxicants in vivo. However, their isolation from testicular homogenates is tedious and requires the regular use of numerous immature animals. It is therefore of great interest to have available established cell lines that are usable in vitro for unlimited periods and closely similar to native cells. To this end, we have established a line of Wistar rat Sertoli cells (SerW3) by immortalization of fresh primary cells with the T antigens of the Simian virus (SV40). When plated on Matrigel, this cell line presents many of the functional characteristics of Sertoli cells in vivo. In addition, they are sensitive to cisplatin and secrete transferrin, although they do not show a clear response to follicle-stimulating hormone. They also present many morphological similarities, including the presence of tight junctions which mimic the natural epithelial barrier. Like Sertoli cells in vivo, they show extensive phagocytic activity. Finally, they display all the characteristics of immortalized, but not transformed, cells, i.e., topo inhibition and apoptosis at confluence or under serum deprivation. PMID- 9352124 TI - Low and very low density lipoprotein composition and resistance to copper-induced oxidation are not notably modified in smokers. AB - To study whether tobacco use was associated with oxidative phenomena affecting lipoproteins, we estimated susceptibility of LDL and VLDL to an in vitro copper mediated oxidation, and measured serum autoantibody titers against oxidized LDL in 45 middle-age healthy nonsmokers, 35 smokers and 37 ex-smokers of both sexes, taking into account the detailed lipid composition of the lipoproteins. VLDL from female smokers had higher triglyceride, phospholipid, apolipoprotein E and alpha tocopherol content and showed a higher rate of copper-induced oxidation in comparison with those from nonsmokers (P < or = 0.05) whereas the relative composition of these particles in saturated, mono- or poly-unsaturated fatty acids was not modified by tobacco consumption. After adjustment for triglyceride content, no statistically significant difference in oxidation rate was observed. Lipid, alpha-tocopherol and protein composition of LDL did not appear to be influenced by smoking; in accordance with these observations, no difference in indices of in vitro oxidizability of LDL was noticed between the different groups. Autoantibody titers against oxLDL were similar in smokers and nonsmokers. We conclude that, in supposed healthy individuals, smoking does not seem to be associated with notable variations in composition of VLDL and LDL or with an increase of oxidizability of these atherogenic lipoproteins. PMID- 9352125 TI - Characterization of chymase from human vascular tissues. AB - A chymostatin-sensitive angiotensin II-generating enzyme was found in human gastroepiploic arteries. The enzyme was purified using heparin affinity and gel filtration columns. The molecular mass of the purified enzyme was 30 kDa, and the optimum pH was between 7.5 and 9.0. Enzyme activity was inhibited by soybean trypsin inhibitor, phenylmethylsulfonyl fluoride and chymostatin, but not by ethylenediaminetetraacetic acid, pepstatin and aprotinin. The enzyme rapidly converted angiotensin I to angiotensin II (K(m), 67 mumol/l; Vmax, 43 pmol/s, kcat, 65/s), but did not hydrolyse angiotensin II, substance P, bradykinin, vasoactive intestinal peptide, luteinizing hormone-releasing hormone, somatostatin and alpha-melanocyte-stimulating hormone. The N-terminal sequence was identical to the sequence for human skin/heart chymase. Thus, the chymostatin sensitive angiotensin II-generating enzyme in human vascular tissues is identified as chymase. PMID- 9352126 TI - Serum laminin and type III procollagen in chronic hepatitis C. Diagnostic value in the assessment of disease activity and fibrosis. AB - Laminin P1 (pepsin-resistant fragment of laminin) and aminoterminal peptide of type III procollagen are measurable in serum and are now considered useful serum markers of fibrogenesis and inflammation in chronic liver diseases. However, very few studies thus far have focused on assessing the diagnostic value of these markers in detecting fibrosis and necro-inflammatory activity in chronically diseased liver. The aim of the present study was therefore to investigate the correlations of laminin and type III procollagen with liver histology and to compare their diagnostic value in detecting the degree of liver fibrosis and necro-inflammatory activity in a homogeneous group of 99 patients suffering from chronic hepatitis C, and lacking other factors which can directly affect the serum levels of the two markers. Both these serum markers were measured by radioimmunoassay, employing commercially available kits. The three main aspects of liver pathology, i.e. portal-periportal activity, lobular activity and fibrosis, were histologically evaluated and semiquantitatively expressed by numerical scores. The results of this study show that laminin and type III procollagen were both positively correlated with the histological scores for portal-periportal activity and with those for fibrosis, whereas no significant correlation was observed between each of the two serum markers and the histological scores for lobular activity. The sensitivity and specificity of laminin and type III procollagen in detecting histological aspects of fibrosis and disease activity in liver, computed at various cut-off levels, showed overlapping trends for the two markers; however, the diagnostic value was in general rather low, whatever the cut-off considered. We therefore conclude that the 'static' measurement of both serum laminin and type III procollagen is of limited value for individual diagnosis of liver damage. PMID- 9352127 TI - The use of myoglobin/carbonic anhydrase III ratio as a marker for myocardial damage in patients with renal failure. AB - To evaluate the clinical significance of myoglobin and myoglobin/CA III ratio as a biochemical marker for acute myocardial infarction (AMI) in patients with renal failure; we studied 300 patients admitted to the hospital with a history of symptoms characteristic of AMI, and 33 renal failure patients who were undergoing chronic maintenance dialysis treatment and who did not have clinical or electrocardiographic evidence of AMI. Fifteen of 300 patients admitted to the hospital had AMI based on the WHO criteria, and a concomitant value of serum creatinine concentration (S-Crea) over 140 mumol/l indicating renal failure. Fourteen of these 15 patients (93%) had serum myoglobin concentration over 70 micrograms/l and myoglobin/CA III ratio over 2.20 as measured by time-resolved fluoroimmunoassay (TR-FIA); these values were cutoff values for AMI diagnosis. Twenty-two of 300 patients admitted to the hospital had S-Crea over 140 mumol/l in the absence of myocardial injury. Sixteen of these 22 (73%) patients had increased serum myoglobin concentration, but only four of 22 (18%) had myoglobin/CA III ratio over 2.20. A positive correlation between serum myoglobin and CA III concentrations (rs = 0.933, P < 0.001) was observed in hemodialyzed patients with chronic renal failure. The values for serum myoglobin/CA III ratio observed in this group were similar to those measured in the 22 non-AMI patients with S-Crea over 140 mumol/l admitted to the hospital and differed statistically from that for patients with AMI (P < 0.001). We conclude that serum myoglobin, as well as CA III values, are elevated in patients with renal failure, and therefore S-myoglobin can not be used as a marker for AMI in these patients. Our results suggest that the serum myoglobin/CA III ratio is a reliable AMI marker even in renal failure patients, and therefore provides a tool for AMI diagnosis in this patient group. PMID- 9352129 TI - The tumor-derived fetal-intestinal alkaline phosphatase cDNA is identical in sequence to the adult intestinal alkaline phosphatase isozyme gene. AB - The alkaline phosphatase (AP) of Caco-2 cells, a cell line derived from a human adenocarcinoma of the colon, is quite similar to fetal intestinal AP in its enzymatic properties. The nucleotide sequence of a cDNA encoding AP produced in Caco-2 cells was examined. The sequence was identical to one of the three sequences of adult intestinal AP reported previously. We further investigated the entire nucleotide sequence of cDNA of intestinal-type AP produced in cancer cell lines such as HuH-7 cells, FL-amnion cells, and HuG-1 cells. The sequence of these cell APs was identical to that of Caco-2 cell AP. These results indicate that cancer cells producing intestinal-type AP have the same nucleotide sequence as that of adult intestinal AP, and suggest that the differences in electrophoretic mobilities of these cell APs compared with adult intestinal AP may be due to post-translational modifications. PMID- 9352130 TI - Prostate-specific antigen (protein and mRNA) analysis in the differential diagnosis and staging of prostate cancer. AB - We analyzed complexed and free prostate-specific antigen (PSA), the free/total PSA and complexed/free PSA ratios, acid phosphatase, and prostatic phosphatase in serum from 36 patients with prostatic carcinoma and from 48 non-neoplastic control patients (20 with prostatitis and 28 with benign prostatic hyperplasia). Receiver-operating characteristic plots showed that serum PSA was the most efficient variable, singly used, in discriminating neoplastic from non-neoplastic patients. At a cut-off value of 10.0 ng/ml, serum PSA had a diagnostic sensitivity of 87% and a diagnostic specificity of 83%. In particular, three patients with prostatic carcinoma and twenty non-neoplastic controls had serum PSA levels of between 4 and 10 ng/ml. The subsequent analysis of the serum free/total PSA ratio, in this subgroup, using a cut-off level of 15%, allowed us to classify correctly all prostatic cancer cases and 18/20 non-neoplastic diseases. We next analyzed PSA mRNA in circulating cells using an improved reverse-transcriptase polymerase chain reaction dot blot procedure, from six patients with prostatic carcinoma with distant metastases, and in seventeen with localized cancer. The analysis had a high sensitivity (up to dilutions 1:10(6) of total RNA from prostatic cancer cells vs total RNA from normal blood cells). The analysis revealed circulating micrometastatic cells in 3/6 (50%) cases of metastatic cancer and in 4/17 cases of localized cancer. To conclude, serum total PSA combined with the free/total PSA ratio is a very efficient algorithm in discriminating neoplastic from non-neoplastic prostatic diseases, while other mRNA species must be analyzed, in addition to PSA mRNA, in circulating cells to increase the efficiency in detecting metastatic prostatic cancer. PMID- 9352128 TI - Overexpression of calcium-binding protein calgranulin B in colonic mucosal diseases. AB - Subtractive two-dimensional gel electrophoresis (2-DE) has been used for the study of the protein patterns of the normal colonic mucosa and the specimens collected from patients diagnosed for inflammatory bowel disease (IBD), colonic polyps and colorectal cancer. We found a 13 kDa protein that was detected in five of seven adenomas and in 13 of 15 colorectal carcinomas while it was absent or only slightly expressed in normal colonic mucosa. Furthermore, this protein occurred in all specimens collected from patients suffering from IBD and its quantity reflected the increased severity of inflammation. The combination of microsequencing and mass spectrometry led to the identification of the 13 kDa spot as calgranulin B. Our results indicate that the production of calgranulin B is unregulated in inflammatory, preneoplastic and neoplastic lesions of colonic mucosa. PMID- 9352131 TI - Lipid peroxidation and antioxidant status in experimental animals: effects of aging and hypercholesterolemic diet. AB - Effects of aging and hypercholesterolemic diet on lipid peroxidation and antioxidant status were investigated in rats. The rats were divided into four groups of ten: Group I; young rats receiving standard lab chow; Group II; young rats on hypercholesterolemic diet (0.4 g/rat/day); Group III; aged rates receiving standard lab chow; Group IV; aged rats on hypercholesterolemic diet (0.4 g/rat/day). Plasma lipid peroxidation end product level was determined as thiobarbutiric acid reactive substances (TBARS). Plasma cholesterol concentration was analyzed by a kinetic enzymatic method. Erythrocyte superoxide dismutase (CuZn SOD), glutathione peroxidase (GSH Px) and glutathione (GSH) levels were determined spectrophotometrically. Cholesterol values were found to be significantly high (p < 0.001), TBARS (0.05 > p > 0.02) and GSH (p < 0.001) levels significantly low in aged rats in comparison with young rats. Hypercholesterolemic diet induced significant increases in GSH (p < 0.001) and CuZn SOD (p < 0.001) levels, whereas a significant decrease in GSH Px activity (0.05 > p > 0.02) was observed in aged rats. In young rats hypercholesterolemic diet caused a significant increase in both GSH and CuZnSOD levels. Our results indicate an imbalance between radical production and destruction in favour of prooxidant conditions in the young rats and the induction by hypercholesterolemic diet of the antioxidative response in erythrocytes. PMID- 9352132 TI - 1H NMR spectra of normal urines: reference ranges of the major metabolites. AB - Serial urine samples from 50 normal subjects were studied by 1H NMR spectroscopy operating at 300 MHz. Analyses of the spectra have shown the presence of the following metabolites in 100% of the normal subjects: Creatinine, lactate, alanine, citrate, dimethylamine, trimethylamine-N-oxide, glycine and hippurate. Other analytes, such as creatine, valine, betaine, leucine and isoleucine, were sometimes found. All metabolites were quantified on the basis of peak heights and were expressed as mmol/mol of creatinine. The study of metabolic profiles in serial samples allowed us to evaluate intra-individual variability and physiological changes due to feeding. The aim of our report is to define standard conditions for this analytical technique and to calculate confidence intervals for the major metabolites in normal urine samples, such as preliminary and mandatory stages for clinical diagnostic 1H NMR utilization. PMID- 9352133 TI - Lipoprotein (a) phenotype distribution in a population of bypass patients and its influence on lipoprotein (a) concentration. AB - A case control study was undertaken to compare the distribution of apolipoprotein (a) phenotypes in patients suffering from atherosclerosis and undergoing coronary bypass surgery with the distribution observed in adequately selected controls. Cases differed from controls for triglycerides (1.90 +/- 0.88 mmol l-1 and 1.16 +/- 0.79 mmol l-1, P < 0.0001, respectively), HDL cholesterol (1.15 +/- 0.34 mmol l-1 and 1.69 +/- 0.42 mmol l-1, P < 0.0001, respectively), apolipoprotein AI (1.31 +/- 0.24 g l-1 and 1.70 +/- 0.29 g l-1, P < 0.0001, respectively) and lipoprotein a (Lp(a)) (0.32 +/- 0.30 g l-1 and 0.19 +/- 0.20 g l-1, P < 0.0001, respectively). The apolipoprotein (a) phenotypes were distributed differently in cases and controls (chi 2 = 25.26, P < 0.0001) with a lower percentage of isoforms of larger size and a higher percentage of isoforms of smaller size in patients. The Lp(a) concentration remained significantly higher in patients than in controls for most of the phenotypes, suggesting that both a high Lp(a) concentration and a different apolipoprotein (a) size distribution could be involved in the development of atherosclerosis in this population. In addition, patients exhibiting the highest Lp(a) concentrations had higher levels of LDL cholesterol and apolipoprotein B than patients exhibiting the lowest Lp(a) concentrations. This feature was not observed in controls. By contrast, controls with the highest Lp(a) concentration had significantly higher triglyceride levels than controls with the lowest Lp(a) concentration. This feature was not observed in patients. Our results indicate that patients undergoing bypass surgery have higher Lp(a) concentrations than controls, this increase being not completely explained by the difference in apolipoprotein (a) phenotype distribution. The high Lp(a) concentration seems to be associated with different lipid profiles in patients than in controls. PMID- 9352135 TI - Quantitation of plasminogen epitopes of serum lipoprotein(a) by sandwich enzyme linked immunosorbent assay. PMID- 9352134 TI - Glutathione and neonatal lung disease. AB - Glutathione (GSH) was measured using HPLC-electrochemical detection in bronchoalveolar lavage fluid from 28 neonates for up to 21 days after birth. GSH levels varied from 0.1-11.2 mumol l-1 (with a geometric mean concentration of 1.3 mumol l-1). GSH in epithelial lining fluid was estimated using the urea dilution method at 15.0 mumol l-1 (range 0.5-196 mumol l-1), which is significantly lower than observed in adult subjects. There was an L shaped relationship between GSH and the two markers of oxygen therapy, oxygen index and FiO2. The lowest GSH levels were associated with the group of infants with the most severe airways problems who required high oxygen. PMID- 9352136 TI - Oxidative damage in red blood cells of vitamin E deficient patients. PMID- 9352138 TI - CYP2D6 Hhal genotype and the neuroleptic malignant syndrome (NMS) PMID- 9352137 TI - Insulin receptor defects in the erythrocytes of obese Indian women with Acanthosis Nigricans. PMID- 9352139 TI - The laboratory diagnosis of malaria. The Malaria Working Party of The General Haematology Task Force of the British Committee for Standards in Haematology. AB - Audits of malaria diagnosis in the UK have revealed shortcomings. The use of recommended procedures should improve the standard of malaria diagnosis. Both thick and thin films should be examined. Thick films should be stained unfixed with a Giemsa or modified Field's stain. Thin films should be fixed and stained with a Giemsa or a Leishman stain. All films should be examined for an adequate period of time by two observers. In the case of P. falciparum infection parasites should be quantified. Microscopy may be supplemented by an immunological or fluorescence-based method. Slides from all cases in which a diagnosis of malaria is made should be sent to a reference centre for verification. Laboratories should participate in a relevant NEQAS scheme and should take steps to ensure that all those carrying out malaria diagnosis maintain their skills. PMID- 9352140 TI - The use of automated HPLC to detect and quantitate haemoglobins. AB - The introduction of automation for haemoglobinopathy screening is an important advance in technology for haematology laboratories. This paper evaluates the utility of an automated HPLC instrument, the Bio-Rad 'Variant' for the detection and quantitation of the normal haemoglobins (Hb A, A2 and F) and the common abnormal haemoglobins (Hb S, C, DPunjab, E, OArab and Lepore) which need to be evaluated in laboratories undertaking carrier and/or neonatal screening for sickle cell and thalassaemia. The instrument only uses a small amount of whole blood (5 microliters), a 3 mm disc from a Guthrie spot may also be used for analysis of samples from neonates. It uses a 100 place automatic sampler with a cycle time of 6.5 min for adult samples (using the 'Beta Thalassaemia Short' reagent pack) and 3 min for neonatal samples. The automatic sampler also allows samples to be analysed 'out of hours'. A 'STAT'; position allows urgent samples to be analysed before, or during, a routine analytical run. All reagents, other consumables and application notes are provided by the suppliers. Other types of reagent packs, such as the 'Sickle Cell Short' for neonatal screening were not assessed during this study. PMID- 9352141 TI - Seasonal differences in blood cell parameters and the association with cigarette smoking. AB - Seasonal changes in red cell parameters need to be defined in order to properly interpret laboratory results. In this study blood counts were obtained prospectively in 104 healthy men (84 non-smokers and 20 smokers) aged 28-63 years during the summer and winter months. Seasonal changes in plasma volume were also calculated. In healthy non-smokers, their haemoglobin concentration and haematocrit ratio were lower in summer than in winter with a concomitant 5.5% increase in plasma volume. In smokers, there was no change in plasma volume, but haematocrit levels increased in summer. We conclude that both smoking status and seasonal variation should be taken into account in the evaluation of blood count results. Further studies are warranted to determine if our results can be extrapolated to subjects of both sexes and of various ages exposed to different climatic conditions. PMID- 9352142 TI - An evaluation of the Spuncrit infra-red analyser for measurement of haematocrit. AB - An open, single centre study was carried out to evaluate the accuracy of the Spuncrit (Micro Diagnostics, Bethlehem, PA, USA) infra-red analyser which can be used for near-patient testing to measure haematocrit and estimate haemoglobin concentration. The primary comparison was with the Sysmex NE1500 (Tao Medical) analyser situated in the main hospital laboratory. Secondary comparison was with the Ciba Corning 288 (Ciba Corning Diagnostics Ltd, Halstead, UK) blood gas analyser currently used for near-patient testing in the Northern General Hospital. A total of 217 samples from 50 patients was analysed. The Pearson's correlation coefficients for haematocrit and haemoglobin concentration between the Spuncrit and Sysmex NE1500 and between the Spuncrit and Ciba Corning 288 were all close, between 0.85 and 0.92. The method of Bland and Altman was used to assess agreement between the results of the Spuncrit and the Sysmex NE1500. The agreement for haematocrit was good with 2 SD of the Spuncrit results being between -5.66 and +4.42% of the measurement from the Sysmex NE1500. In conclusion, the Spuncrit haematocrit measurement agreed well with results from the central laboratory, but the estimated haemoglobin concentrations agreed less well and three reasons are discussed. PMID- 9352143 TI - Evaluation of Sysmex SF-3000 performance concerning interpretive morphology flagging of the leucocyte differential count. AB - Results from the Sysmex SF-3000 automated haematology analyser are reported with special attention to leucocyte morphology flagging in pathological conditions. Results for leucocyte differential counts provided by the Sysmex SF-3000 were compared with those obtained from the Sysmex NE-8000 and a visual evaluation based on a differential count of 400 leucocytes. One hundred samples from apparently healthy subjects and 100 samples from patients with haematological abnormalities were examined. The SF-3000 yielded a rather high frequency of flagging inappropriately referring to blasts, immature granulocytes, left shift and nucleated red blood cells/platelet clumps. The presence of atypical lymphocytes was not indicated by appropriate flagging. PMID- 9352144 TI - An evaluation of the differential from the Abbott CD 3500 in a population of patients with haematological abnormalities. AB - The Cell Dyn 3500 (CD 3500) Haematology Analyser (Abbott Diagnostics, Lane Cove, NSW, Australia) has been evaluated to determine the reliability of the differential and white cell suspect flagging in a population of patients with known haematological abnormalities. The evaluation included the assessment of white cell differential parameters in addition to sensitivity, specificity and efficiency of white cell suspect flagging. The study showed that the CD 3500 is an efficient and sensitive screening tool for detecting the presence of clinically significant white cell abnormalities. Generally, the Blast and Band flags demonstrated the highest level of false positive flagging (lowest sensitivity). The immature granulocyte flag was found to be an extremely reliable indicator of the presence of myelocytes, metamyelocytes and promyelocytes on the film. There was a 0.4% false negative rate where significant numbers of variant lymphocytes (> 10%) were not detected by the CD 3500 on flagging alone. When analyser flags and white cell scatter distribution are considered in combination with defined laboratory limits, all white cell suspect flags have acceptable sensitivity, specificity and efficiency rates. PMID- 9352145 TI - Serum levels of tumour necrosis factor-alpha predict response to recombinant human erythropoietin in patients with myelodysplastic syndrome. AB - We measured pretreatment serum levels of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) in 25 patients with myelodysplastic syndrome receiving recombinant human erythropoietin (rhEPO) at dosages up to 300 U/kg thrice weekly for 12 weeks. Both TNF-alpha and IL-1 beta levels were measured using commercially available enzyme-linked immunoassays. A complete response (CR) was defined as a rise in untransfused haemoglobin concentrations of at least 2 g/dl or a 100% decrease in RBC transfusion requirements over the treatment period; a partial response (PR) was an increase in untransfused haemoglobin values of 1-2 g/dl or a decrease in RBC transfusion requirements equal to or greater than 50%; no response (NR) was defined as a response less than a PR. After 12 weeks of rhEPO treatment, four patients showed a CR, five patients a PR, and 16 patients NR. Serum levels of both TNF-alpha (80.5 %/- 64.8 vs 8.1 +/- 4.2 ng/l, P < 0.001) and IL-1 beta (60.4 +/- 49.9 vs 8.9 +/- 4.7 ng/l, P < 0.001) were higher in MDS patients than in a group of 28 normal controls. Responders (CR + PR) showed significantly lower serum levels of TNF-alpha than non-responders (21.6 +/- 26.2 vs 106.3 +/- 60.8 ng/l, P < 0.001), whereas IL-1 beta concentrations between those who benefited from therapy and unresponsive cases were not significantly different (39.8 +/- 48.9 vs 73.4 +/- 48.2 ng/l, P = 0.120). It is noteworthy that TNF-alpha levels were within the normal range in all responsive patients but one, whereas all non-responders presented elevated cytokine concentrations. No relationship was found between TNF-alpha or IL-1 beta values and haemoglobin levels, transfusion requirement, serum EPO or ferritin concentrations. We conclude that pre-treatment TNF-alpha levels might help to select those MDS patients who are most likely to benefit from rhEPO treatment. PMID- 9352146 TI - Comparison of oral anticoagulant control by a nurse-practitioner using a computer decision-support system with that by clinicians. AB - With increasing work-loads in anticoagulant clinics different methods of service delivery need evaluation. The quality of anticoagulant control achieved by a nurse-practitioner using a computer decision-support system (CDSS) was compared with that achieved by trainee doctors without CDSS. Eighty-one out-patients (group A, therapeutic range 2-3) and 96 out-patients (group B, therapeutic range 3-4.5) were randomized to management by a nurse-practitioner or by trainee doctors (clinicians). Thirty-seven patients in group A and 50 patients in group B were randomized to be managed by the nurse-practitioner. In group A, patients in the nurse-practitioner group spent a longer time in the therapeutic range than those in the clinician group (60.7% compared with 51.6%). Dose suggestion acceptance in the nurse-practitioner group (88%) was higher compared with agreement between the CDSS and the clinicians (60%). In group B, patients in the clinician group spent a slightly longer time in the therapeutic range (70% compared with 67.6%). Acceptance of dose suggestion was lower in the nurse practitioner group (67%) compared with agreement between the CDSS and the clinicians (73%). In conclusion, the CDSS can improve the quality of control of warfarin therapy by a nurse-practitioner over that by trainee doctors for the therapeutic range 2-3. Similar quality of control is achieved for the therapeutic range 3-4.5. The CDSS may be used by nurse-practitioners to achieve safe and effective anticoagulation in hospital-based or out-reach anticoagulant clinics. PMID- 9352148 TI - Priapism in a patient with protein C deficiency. AB - We present the case of a patient with classical protein C deficiency presenting with acute priapism during warfarinization for thrombophlebitis. Priapism is a well-recognized complication of a number of conditions including sickle cell disease and haematological malignancies, but to our knowledge it has not previously been reported in association with protein C deficiency. This case highlights the potential dangers of initiating oral anticoagulant therapy using conventional loading dose regimens in patients with protein C deficiency. PMID- 9352147 TI - An original method to study autoantibody specificity in haemoglobin stained eluates by the column agglutination techniques. AB - When studying autoantibody specificity by the indirect antiglobulin test with column agglutination techniques ether and xylene elution techniques result in haemoglobin stained eluates which give a red colouration to the gel or glass beads and do not allow the identification of positive reactions. Xylene eluates were incubated with commercially available group O-test red cell panels at 37 degrees C for 45 min in the wells of a microtitre plate in a 3:1 eluate:red cell ratio. After washing with normal saline, sensitized red cells, resuspended in low ionic strength solution (LISS), were applied onto the microtubes containing the antiglobulin serum and positive reactions were recorded after centrifugation. We studied the specificity of 35 autoantibody containing eluates from 12 patients with lymphoproliferative disorders (six having autoimmune haemolysis) and 23 HIV patients without autoimmune haemolysis. All patients had a gel or column positive (IgG) direct antiglobulin test while the tube direct antiglobulin test failed to show red cell bound IgG. We found a reactive indirect antiglobulin test in 20/23 eluates from HIV infected patients (with a panreactive specificity), in all patients with autoimmune haemolysis (one with anti-C, two with anti-E, one with anti-K and two with a panreactive specificity) and in all patients with positive direct antiglobulin test but without immune mediate haemolysis (in all cases with panreactive specificity). The method proposed is a promising tool for the study of the specificity of antibody containing haemoglobin stained eluates; in this study it allowed us to confirm that some HIV patients have specific binding of IgG on their RBC and to identify the specificity of tube test non-reactive eluates. PMID- 9352149 TI - Interaction of cyclosporin A and etoposide. Clinical and in vitro assessment in blast phase of chronic myeloid leukaemia. AB - Combination chemotherapy has had a low impact on survival of blast crises in chronic myelogeneous leukaemia (CML) which may be due to drug resistance. This work attempted to correlate the clinical response and some experimental evidence for the MDR phenotype. Blast cells were positive for P-glycoprotein using APAAP assay. In vitro tests showed that etoposide was partially toxic to blast cells when used alone but had its toxicity increased by nearly sixfold when combined with cyclosporin A (CSA). The patient responded poorly to treatment with etoposide combined with mitoxantrone and high-dose ara-c. However, when etoposide was associated with CSA, this patient returned to the chronic phase reinforcing our in vitro studies. Because no serious toxicity was seen clinically, we are inclined to consider the circumvention protocol an useful strategy to treat blast crises of CML. PMID- 9352150 TI - Sea-blue histiocytosis and pancytopaenia associated with chronic total parenteral nutrition administration. AB - A 22-year-old female on chronic total parenteral nutrition for short bowel syndrome presented for investigation of pancytopaenia and hepatosplenomegaly. Bone marrow examination revealed an infiltrate of sea-blue histiocytes and cytochemistry confirmed these to be lipid laden macrophages. The total amount of fat in the feeding regimen was subsequently reduced, and there has been a partial haematological improvement. The occurrence of sea-blue histiocyte syndrome complicating the fat emulsion component of chronic total parenteral nutrition has been reported recently. To our knowledge this report is the first where reduction in the lipid content of the feeding regimem has resulted in an improvement in the degree of pancytopaenia. PMID- 9352152 TI - Neutrophil leucocytosis in atherosclerosis. PMID- 9352151 TI - Autoimmune haemolytic anaemia and thrombocytopenia associated with ciprofloxacin. PMID- 9352153 TI - Gene-polymorphisms of angiotensin converting enzyme and endothelial nitric oxide synthase in patients with primary glomerulonephritis. AB - The ACE D- and the ecNOS a-allele have been associated with an adverse prognosis in patients with glomerulonephritis (GN). Using genomic DNA we investigated by RT PCR whether the two polymorphisms are useful prognostic markers in GN patients. In patients with primary GN (IgA-GN n = 70, membranous GN n = 23, FSGS n = 17, MPGN n = 6) neither the whole group nor disease-specific subgroups exhibited any alterations from the normal ACE genotype distribution. No significant associations were detected between the ACE genotype and the development of hypertension, antihypertensive therapy required, progression rate of the disease, age of diagnosis and the antiproteinuric response to ACE-inhibition. In 40 IgA-GN patients with ESRD no increased prevalence of the D-allele was noted. The distribution of the ecNOS-alleles in the above patients (a-allele 22%, b-allele 78%) was comparable to that of the normal controls. No association with any of the parameters mentioned above were detected in the case of the ecNOS-alleles. CONCLUSIONS: In our Caucasian patients neither the determination of the ACE nor the ecNOS genotype offered any diagnostic or prognostic help. PMID- 9352154 TI - Decreased excretion of glycosaminoglycans in patients with primary glomerular diseases. AB - Urine glycosaminoglycans (GAG) concentrations were measured in 150 patients with primary glomerulonephritides: endocapillary glomerulonephritis, mesangial proliferative glomerulonephritis, IgA nephropathy, membranous glomerulonephritis and minimal change nephropathy, and in 63 healthy controls and 19 patients with diabetes nephropathy. The urine GAG to creatinine ratios (GCR) were significantly reduced (p < 0.01) in all the glomerulonephritides investigated (0.20 mg/mmol in endocapillary glomerulonephritis, 1.60 mg/mmol in mesangial proliferative glomerulonephritis, 1.74 mg/mmol in IgA nephropathy, 1.09 mg/mmol in membranous nephropathy, and 1.16 mg/mmol in minimal change nephropathy) compared to healthy controls (2.87 mg/mmol) but not compared to diabetes patients (1.17 mg/mmol). Also, the GCR in a group of 23 non-albuminuric glomerulonephritis patients (1.98 mg/mmol) was shown to be significantly decreased (p < 0.01) compared to healthy controls. Moreover, the GCR was significantly lower (p < 0.01) in endocapillary glomerulonephritis than in any of the other diseases studied. The GAG excretion per functioning glomerular area, calculated as fractional GAG excretion (FGE), was decreased in all the glomerulonephritides investigated compared to both healthy controls and diabetes nephropathy. The decreased GAG excretion in glomerulonephritides, obtained in the present study, might be a consequence of decreased synthesis or turnover of GAG in the functioning nephrons whereas the mechanisms for the reduced GAG excretion in diabetes nephropathy might be of a different nature. Urinary GAG excretion in this group of glomerular disorders and particularly in endocapillary glomerulonephritis, may lead to new approaches in non-invasive renal diagnostics and, particularly with regard to the differentiation of acute and chronic forms of glomerulonephritides. PMID- 9352155 TI - Renovascular hypertension may cause nephrotic range proteinuria and focal glomerulosclerosis in contralateral kidney. AB - Little attention has been paid to nephropathies and proteinuria in renovascular hypertension (RVH). Recently there has been a growing interest in the conditions induced by RVH. 10 cases of RVH were diagnosed by angiography and renin sampling from renal veins in the last 6 years in our hospital. The patients were all male and mean age was 64 +/- 8 (SD) years. Data were as follow: protein excretion was 3.8 +/- 2.2 g/day (> or = 3.5 g/day in 8 patients), sBP 202 +/- 24 mmHg, dBP 113 +/- 17 mmHg, serum renin concentration 64 +/- 45 pg/ml, and ipsilateral/contralateral renal vein renin ratio 3.3 +/- 1.0. RVH was treated by nephrectomy in 3 patients, percutaneous transluminal renal angioplasty (PTA) in 2, and angiotensin converting enzyme inhibitors (ACE-I) administration in 8. Biopsies were performed on contralateral kidney in 4 patients. Focal segmental glomerulosclerosis (FGS) was found in 3 patients, and nephrosclerosis in 1, whereas only nephrosclerosis was found in nephrectomized kidneys in all 3 patients. After nephrectomy, PTA and the treatment by ACE-I, not only blood pressure but also proteinuria was markedly reduced. These findings suggest that severe stenosis of the renal artery led to renal ischemia, which activated renin excretion, to cause glomerular hyperfiltration through vasoconstriction of the efferent arterioles in the contralateral kidney. FGS-like lesion thus induced appeared to have caused massive proteinuria. PMID- 9352156 TI - Effect of lacidipine, a dihydropyridine calcium antagonist on renal function of hypertensive patients with renal insufficiency. AB - There are few studies on the use of dihydropyridine calcium antagonists in hypertensive patients with moderate renal insufficiency. We undertook an open study on the effects on renal function, albumin excretion and blood pressure of the slow-onset, long-acting dihydropyridine calcium antagonist, lacidipine, in 14 patients with stable, chronic renal insufficiency (mean assessed GFR 0.78 ml/s, range 0.50-1.17 ml/s) and moderate hypertension. Following a 2 week washout phase, lacidipine was administered for 24 weeks in a dose of 2 mg/day with the dose being titrated at 2 weekly intervals to a maximum of 6 mg/day in order to achieve adequate blood pressure control. Frusemide was introduced if blood pressure was not controlled on the maximum lacidipine dose. Blood pressure, creatinine clearance, 24 h urinary albumin excretion and plasma creatinine and albumin concentrations were measured at regular intervals throughout the study. Isotopic GFR was determined at the end of the washout period and at week 24. Lacidipine was not very effective in controlling blood pressure and had an adverse effect on renal function. In 3 patients with an incipient nephrotic syndrome this necessitated withdrawal from the study. Mean GFR of the 10 patients who completed the study decreased from 0.69 ml/s/1.73 m2 at baseline to 0.56 ml/s/1.73 m2 at week 24 (p = 0.006) with a decline in GFR being observed in 9 of these patients. The decrease in GFR was greatest in patients with poorly controlled blood pressure. An insignificant increase in mean urinary albumin excretion occurred during the study with this increase being observed only in patients with albuminuria > 1 g/24 h at baseline. These findings indicated that systemic hypertension altered glomerular hemodynamics and that the vasodilatation of pre-glomerular vessels which followed introduction of the calcium antagonist may have exacerbated this situation. The withdrawal of an angiotensin converting enzyme inhibitor during the washout period may have contributed to these changes. We suggest that renal function should be monitored closely in patients with renal insufficiency when a calcium antagonist is being used to control blood pressure, particularly in those with either marginal blood pressure control, significant albuminuria or an incipient nephrotic syndrome. PMID- 9352157 TI - Correction of acidosis in dialysis patients increases branched-chain and total essential amino acid levels in muscle. AB - Earlier studies have shown increased oxidation of the branched-chain amino acids (BCAA), valine, isoleucine and leucine, in experimental acidosis and low levels of valine in the muscle of acidotic HD patients. Using HPLC, free amino acids in plasma and muscle were studied before and after correction of acidosis in 9 HD patients over 6 months by dialysis with a high bicarbonate solution. The predialysis standard bicarbonate concentration in blood increased from 20.6 +/- 1.3 mmol/l (mean +/- SD) before correction of acidosis to 25.9 +/- 1.8 mmol/l after correction. Correction of acidosis resulted in a significant increase in the i.c. concentrations of valine, isoleucine and leucine by 48%, 28% and 32%, as well as for the sum of BCAA and EAA, from a level lower than controls. The intra- and extracellular gradient increased for several amino acids and for the sum of EAA and BCAA, suggesting an increased influx or reduced efflux of amino acids across the cell membrane. Anthropometric data and the levels of S-albumin and transferrin did not change after correction of acidosis. The increases in the i.c. concentrations of BCAA after correction of acidosis suggest that the catabolism of these amino acids had been reduced. The effects of correction of acidosis on the concentrations of essential amino acids could be beneficial since low concentrations in muscle may reduce protein synthesis. PMID- 9352158 TI - Moderate metabolic acidosis and its effects on nutritional parameters in hemodialysis patients. AB - We screened the laboratory data of all of our chronic hemodialysis patients to identify 2 groups: 1) those with a pre-dialysis total CO2 concentration equal or less than 21 mEq/l (group A) and 2) those with a pre-dialysis total CO2 concentration equal or greater than 25 mEq/l (group B) and then both groups were compared for the following parameters: protein catabolic rate, dietary protein intake by dietary history, Kt/V, serum albumin, weight, pre-dialysis serum creatinine and pre-dialysis BUN. Patients from group A had a significantly lower age, a significantly higher protein catabolic rate and significantly higher values for pre-dialysis serum creatinine and BUN. The values for body weight, dietary protein intake and serum albumin were also higher in group A than in group B but the differences did not reach statistical significance. There was a good correlation between protein catabolic rate and pre-dialysis total CO2 and between the latter and serum albumin. These results suggest that moderately low pre-dialysis serum bicarbonate concentration is usually the result of high protein intake and should be of no concern in well-dialyzed patients with a protein catabolic rate greater than 1 g/kg/day. However, further studies are needed to confirm this conclusion. PMID- 9352160 TI - Cardiac surgery in patients with end-stage renal failure. AB - End-stage renal failure is commonly considered a significant factor for an increased risk after coronary artery bypass grafting. This holds true for patients who have received a kidney transplant (NTX group) as well as for patients who require chronic hemodialysis (HD group). To assess the risk in our population we performed a retrospective analysis of 22 patients with end-stage renal failure (HD group: 17, NTX group: 5) who underwent cardiac surgery. The perioperative course was compared to a normal population. In addition to standard data we assessed the following factors: renal failure etiology, risk factors, concurrent diseases, duration of renal failure, function of renal graft, ECG (paying special attention to signs of previous myocardial infarctions and rhythm disorders), results of cardiac catheterization and coronary angiography, NYHA class and urgency of operative intervention. Complications and mortality were the main measures of the perioperative course. We analyzed the hospital charts retrospectively and requested the patients' physicians to complete a questionnaire about the patient's present condition. All HD group patients were dialyzed on the day before surgery. The first postoperative HD was performed for hyperkalemia or signs of volume overload (pulmonary capillary wedge pressure > 20 mmHg) when signs of pulmonary function deterioration were seen. HD was successful in treating these conditions. 3 of the 17 patients on HD expired postoperatively, 4 died within 3 years, all of unrelated diseases. Mortality and morbidity was 0% in the NTX group. In one NTX patient who required intermittent HD preoperatively because of poor renal graft function, renal function improved postoperatively, presumably secondary to better renal perfusion, and he did not require HD after his cardiac surgery. By surgical intervention the NYHA class of all patients improved (by 1.6 on the average) as well as their quality of life. Because of these good short- and long-term results and relatively low operative risk we support an approach of prompt work-up and surgical intervention when necessary in HD and NTX patients. PMID- 9352159 TI - Increase urinary hepatocyte growth factor excretion in human acute renal failure. AB - Studies in animals suggest that hepatocyte growth factor (HGF) is an important mediator of kidney development, compensatory growth and tubule repair following acute injury, however, evidence for HGF action in human renal disease is scant. To determine whether increased renal production of HGF occurs in man, urine HGF excretion rate was measured in normals and in patients with a variety of acute and chronic renal diseases. Urine samples were collected from 9 healthy individuals, 25 individuals with acute tubular necrosis (ATN), 20 individuals with chronic glomerular disease, 9 patients with polycystic kidney disease and 10 individuals with severe chronic renal failure not yet receiving renal replacement therapy. Samples were initially frozen and then HGF content measured by ELISA and factored for creatinine concentration measured by autoanalyzer. Detectable but low levels of HGF were found in the urine of normals and in patients with chronic glomerular or polycystic disease. Levels were also not increased in patients with advanced, chronic renal insufficiency. In contrast, a marked increase in urine HGF was observed in patients with acute renal failure. In addition, HGF excretion tended to correlate with disease severity as higher levels were observed in patients with oliguric ATN. Urine HGF levels declined to control values in patients recovering from ATN, generally within one week. These findings are consistent with a role for HGF in promoting tubule cell proliferation, differentiation and recovery from acute tubular injury in man. PMID- 9352161 TI - Clinical characterization of Dicea a new cellulose membrane for haemodialysis. AB - A prospective randomised clinical study comparing the functional performance and biocompatibility of a new cellulose diacetate variant (Dicea) in which the degree of hydroxyl group substitution differs, with cellulose diacetate and low flux polysulfone incorporated into commercially produced hollow fiber hemodialysers with a surface area 1.5-1.6 m2 has been undertaken. All dialysers studied demonstrated clinically acceptable performance in terms of their small molecular removal characteristics, with minor statistical but not clinical differences. Use of both cellulose diacetate membranes but not low flux polysulfone resulted in a reduction in plasma beta(2) microglobulin levels. The membranes were impermeable to albumin, but showed some permeability to low molecular weight proteins. The average protein recovery from the dialysis fluid was 3105 mg for Dicea, 2913 mg for cellulose diacetate and 2842 mg for low flux polysulfone. For Dicea the white cell count by 15 minutes had declined to 68% of pre treatment value, compared with 59% and 86% for cellulose diacetate and low flux polysulfone. The differences between Dicea and cellulose diacetate were not significant, but both cellulose based membranes differed from low flux polysulfone (p = 0.0015). There was a strong evidence of differences between the membranes in respect of C5a and C5b-9 generation (p = 0.0001) but not for C3a (p = 0.16) furthermore the levels of C5b-9 generated during dialysis also showed a significant positive correlation compared to C5a for all membranes. (Pearson's correlation coefficient = 0.856, p = 0.0001). It is concluded that the two cellulose diacetate membranes are not identical, with the differences observed being a consequence of the degree of acetyl substitution, resulting in alteration of membrane structure and the method of sterilization. The clinical significance of these differences are difficult to characterize but the modification of the cellulose structure appears to be a promising method to improve the biocompatibility of cellulose membranes. The improved biocompatibility offered by this method still falls short of that achieved with low flux synthetic membranes such as Fresenius Polysulfone. PMID- 9352162 TI - Acute renal failure after ingestion of Cortinarius speciocissimus. AB - In August 1995 a 23-year-old man was admitted to the hospital because of acute anuria. 14 days prior to admission he had consumed five fruit bodies of raw mushrooms of the Cortinarius speciocissimus species. The tentative diagnosis of acute renal failure due to orellanine intoxication was confirmed by the histologic finding of an acute interstitial nephritis in a first renal biopsy one week after onset of anuria. The patient required hemodialysis for the following weeks and months, is now on peritoneal dialysis and is awaiting renal transplantation. Six months after onset of symptoms a second renal biopsy was performed, which revealed increasing interstitial fibrosis. In contrast to the findings of Rapior et al. 1989, orellanine could not be detected in this specimen. The negative toxin test in this second renal biopsy is possibly explained by a wide variability of pharmacokinetics of orellanine. PMID- 9352163 TI - Papillary necrosis: a late complication of nephropathia epidemica? AB - In the following we describe a case of nephropathia epidemica, which exhibited, a few years after acute infection, arterial hypertension and multiple papillary necrosis. Papillary necrosis was diagnosed by i.v. pyelography and CT Scan. A complete evaluation of the patient failed to show any other etiology for medullary necrosis. If arterial hypertension has already been reported as a complication of nephropathia epidemica, papillary necrosis is exceptional, but may be explained by the severe vascular troubles engendered by Hantavirus infection in kidney medulla. PMID- 9352164 TI - Acute hemorrhagic gastritis associated with acetazolamide intoxication in a patient with chronic renal failure. AB - Acetazolamide (Diamox) is a carbonic anhydrase inhibitor commonly used in patients with glaucoma in order to reduce intraocular pressure. Acetazolamide (AZ) is mostly excreted in the urine, therefore, the blood levels of AZ often tend to increase in patients with chronic renal failure. We experienced a case of chronic renal failure in a patient suffering from acute hemorrhagic gastritis associated with AZ intoxication. A 66-year-old female with chronic renal failure was referred to our hospital because of drowsiness and an acute deterioration of renal function. She had been treated with AZ, 500 mg per every day for eleven days for the treatment of glaucoma. Laboratory studies showed leukocyturia, thrombocytopenia, severe anemia, and tarry stools. The serum concentration of AZ was elevated to a maximum of 76.5 mg/ml. She was thus diagnosed as having AZ intoxication. On further examination, acute extensive hemorrhagic gastritis was also found by gastroscopy. Despite of the administration of intensive therapies, she died of disseminated intravascular coagulation (DIC) and septic shock due to bone marrow depression 6 days after admission. It is generally known that excessive blood levels of AZ inhibit not only the gastric juices but also prostaglandin levels and HCO3- excretion in the gastric mucosal barrier. We thus concluded that an excessive dose of AZ had probably destroyed the gastric mucosal barrier or thrombocytopenia due to bone marrow disorder and thus eventually led to the development of hemorrhagic gastritis. As far as we know, this is the first case report of acute hemorrhagic gastritis associated with AZ intoxication. Even though AZ tends to strongly bind to plasma protein and its clearance is generally poor by hemodialysis (HD), in our patient, HD was observed to be rather effective since the clearance of AZ was 45.8 ml/min on HD and 66 ml/min on direct hemoperfusion (DHP). DHP often reduces the number of platelets, also DHP needs a lot of heparin, therefore, we should have performed HD alone instead of DHP. In patients with an impaired renal function, AZ should therefore be administered very carefully in order to avoid an accumulation of the drug. In addition, HD alone should be used to remove any excessive amounts of AZ from the blood. PMID- 9352165 TI - An analysis of the self of the western physician: a study on the evolution of homo Hippocratus. AB - This paper is about western physicians in general, and the Israeli variant in particular. The theoretical focus will be on the nature and dynamics of the medical professional self within this interplay between the cultural concepts of universality and difference. The discussion will address the genesis of this professional self and the implications of its extraordinary strength and tenacity, which has endured for centuries in different local contexts wherever western medicine has existed. The impetus for this study is the subdued rumbling which is echoing throughout the profession suggesting that this self is no longer the "Rock of Gibraltar" that it seemed in the past. PMID- 9352166 TI - For doctors' eyes only: medical records in two Israeli hospitals. AB - "Scientific" and "craft" representations of medical diagnosis can be regarded as complementary discursive systems used by physicians in order to legitimate and monopolize their professional power. This paper examines the medical record as a context for the interplay of these two discourses. During interviews conducted with 78 Israeli physicians, 94% have refused to give patients access to their medical records. This refusal is discussed vis-a-vis a reading of the actual contents of medical records, which are shown to contain many errors, inconsistencies and ambiguities. The paper concludes by offering an alternative, anthropological model for medical records as fieldnotes. PMID- 9352167 TI - The cultural construction of social support in Brazil: associations with health outcomes. AB - The association of social support and health outcomes has received considerable attention in recent years, but the cultural dimension of social support has not been extensively investigated. In this paper, using data collected in a Brazilian city, we present results indicating that those individuals whose reported access to social support more closely approximates an ideal cultural model of access to social support have lower blood pressure and report fewer depressive symptoms and lower levels of perceived stress. The cultural model of social support is derived using a combination of participant observation, semi-structured interviews, and the systematic ethnographic technique of cultural consensus modelling. These results are then used to develop a measure of an individual's approximation to that model of social support in a survey of four diverse neighborhoods in the city (n = 250). We call this approximation to the ideal cultural model of social support "cultural consonance" in social support. The association of health outcomes with cultural consonance in social support is independent of individual differences in the reporting of social support, and of standard covariates. In the case of blood pressure and perceived stress, it is independent of diet, and other socioeconomic and psychosocial variables. The association with depressive symptoms is not independent of other psychosocial variables. The implications of these results are discussed with respect to research on cultural dimensions of the distribution of disease. PMID- 9352169 TI - Cultural formulation of psychiatric diagnosis. Sakit Gila in an Iban longhouse: chronic schizophrenia. PMID- 9352168 TI - Expressions of anxiety in African Americans: ethnography and the epidemiological catchment area studies. AB - High levels of anxiety have long been reported for African Americans. Recent analyses of Epidemiological Catchment Area (ECA) data have failed to support this, although contemporary ethnographies have discussed important African American folk idioms of anxiety. This study compares ethnographically reported symptoms of anxiety in African Americans to those reported in the ECA data. A multivariate analysis of female African American and European American differences in comparable ECA and ethnographic symptoms was performed. Significant differences were found not in ethnicity but in education levels. Alternative interpretations are discussed. Methodological problems are discussed highlighting limitations of both household survey research, such as the ECA project, and ethnography. PMID- 9352170 TI - Transmyocardial laser revascularization for inoperable coronary artery disease. AB - Interest in transmyocardial laser revascularization for the treatment of otherwise inoperable coronary artery disease has increased rather dramatically in recent years. The results of several industrially sponsored clinical series have been reported recently, all with significant improvement in angina pectoris that appears both rapid and sustained. In most instances, an associated improvement in exercise tolerance has been reported. Improvement in regional myocardial perfusion has been proclaimed, although it is less consistent and less complete than symptom relief. The mechanisms whereby this clinical effect is achieved remain unknown. Histologic analysis of autopsy material has yielded somewhat conflicting results regarding the persistent patency of laser-created channels. The results of laboratory investigations of this therapy have been equally inconsistent. Despite our ignorance regarding the mechanism of angina relief, clinical experience continues to grow. In addition to the CO2 laser energy source used in early studies, trials of alternative devices using holmium:yttrium aluminum-garnet and eximer lasers are underway. The latter two employ fiberoptic technology and are currently under development for endovascular approaches. PMID- 9352171 TI - Glycoprotein IIb/IIIa inhibitors in unstable angina. AB - The inhibitors of platelet membrane integrin receptor glycoprotein IIb/IIIa occupy the receptor, preventing fibrinogen binding and platelet aggregation. The inhibition is direct and may be advantageous over the partial inhibition induced by agents interfering at individual pathways to platelet aggregation. The inhibitors, and more specifically abciximab, the monoclonal antibody against the receptor, are effective to prevent the acute complications associated with percutaneous intervention procedures. Based on the positive results of these trials, which have enrolled a large proportion of patients with unstable angina, and on a few pilot studies in unstable angina, large trials have been completed and are ongoing in patients with an acute coronary syndrome. It is hoped that these drugs will prevent the acute complications associated with thrombus formation, and also, past the acute phase, prevent the recurrence of the disease and rapid progression of atherosclerosis. PMID- 9352172 TI - Adjunctive therapy for acute myocardial infarction. AB - Based on results from a multitude of clinical trials of acute myocardial infarction, the beneficial role of a variety of pharmacologic agents, used in conjunction with thrombolytic therapy, has emerged. This is particularly true for aspirin, beta-blockers, heparin, and angiotensin-converting enzyme inhibitors. In addition, the dosage and timing of their administration have been shown to be crucial to their efficacy. The scientific and pathophysiologic basis as well as practical recommendations regarding their use are discussed. The exact role of nitrate therapy and potential for magnesium are presented in addition to data on agents that might be detrimental in the early stages of acute myocardial infarction. PMID- 9352173 TI - Assessment of myocardial viability. AB - The noninvasive assessment of myocardial viability has proved clinically useful for distinguishing hibernating myocardium from irreversibly injured myocardium in patients with chronic ischemic heart disease or recent myocardial infarction who exhibit marked regional and global left ventricular dysfunction. Noninvasive techniques utilized for detection of viability in asynergic myocardial regions include single-photon-emission CT perfusion imaging with 201Tl or one of the new 99mTc-labeled perfusion agents, positron emission tomographic imaging of perfusion and glucose uptake, low-dose dobutamine echocardiography for assessment of inotropic reserve, and contrast echocardiography for evaluation of microvascular integrity. The greater the number of viable myocardial segments by any of these techniques, the greater is the probability of improvement in regional and global left ventricular function, improvement in heart failure symptoms and functional capacity, and enhanced survival after revascularization. Patients with a decreased left ventricular ejection fraction and extensive myocardial viability treated medically have a high cardiac event rate. Similarly, patients with poor viability preoperatively who still undergo coronary bypass surgery have a high rate of early and late cardiac death or need for transplantation compared with patients with greater viability. Finally, some patients with severe ischemic cardiomyopathy referred for cardiac transplantation may have substantial zones of hibernation and may still be candidates for coronary bypass surgery, even in the absence of angina. PMID- 9352174 TI - Intracoronary irradiation for the prevention of restenosis. AB - Restenosis after coronary angioplasty is a major limitation of an otherwise highly effective and safe procedure for the treatment of atherosclerotic coronary artery disease. Although the advent of coronary stenting has reduced restenosis rates for selected patients, an overall restenosis rate of 20% to 25% remains. Despite numerous trials, no effective pharmacologic therapy has been found. Intracoronary irradiation is a new technique proposed to prevent restenosis after angioplasty. In animal models of restenosis after balloon injury, there is marked reduction of neointimal proliferation when the injured vessel is irradiated, using a variety of radiation sources and delivery systems. Early human trials have underscored the importance of careful source calibration and dosimetry. A small, randomized, double-blind, placebo-controlled study of intracoronary irradiation to prevent recurrent restenosis recently reported striking reductions in angiographic restenosis as well as clinical event rates. A number of important issues remain unresolved, such as defining which component of the arterial wall serves as the target tissue for radiation, the minimal effective dose, the maximum tolerable dose, and user-friendly radiation delivery systems. Further studies are needed to define the safety, efficacy and the ultimate usefulness of intracoronary irradiation as an adjunct to current procedures in interventional cardiology. PMID- 9352176 TI - Minimally invasive coronary artery bypass grafting. AB - Minimally invasive cardiac surgery has generated a tremendous amount of enthusiasm in the cardiology and cardiac surgical communities. Coronary revascularization without cardiopulmonary bypass through a small anterior thoracotomy or mediastinotomy has been introduced as an alternative to the conventional approach. An endovascular or port-access technique for cardiopulmonary bypass and cardioplegic arrest has been developed for use in cardiac surgery. This peripherally based system achieves aortic occlusion, cardioplegia delivery, and left ventricular decompression; thus, coronary revascularization and various cardiac procedures can be effectively performed in a less invasive fashion than conventional median sternotomy. Continued technical advances in minimally invasive cardiac surgery will facilitate these procedures, increase patient safety, and contribute to acceptable long-term results. PMID- 9352175 TI - Ischemic preconditioning. AB - It has been shown that repeated brief coronary occlusions increase myocardial resistance towards prolonged episodes of ischemia. This phenomenon, which renders the heart more tolerant to ischemia with subsequent limitation of infarct size, has been termed ischemic preconditioning and has been described in a variety of species. Preconditioning may also protect the heart against postischemic dysfunction and ventricular arrhythmias. Although the beneficial effects seem to be transient, they re-appear at 24 hours, representing a "second window of protection." Ischemia-induced activation of adenosine receptors and opening of ATP-sensitive potassium channels appear to play a role in the acute cardioprotection. For the late protection, stress protein synthesis may play a role. There is experimental and clinical evidence that preconditioning effects may also exist in the human heart. If so, the mechanisms of ischemic preconditioning might be applied to future therapy. PMID- 9352178 TI - Hypertension. PMID- 9352179 TI - Diseases of the aorta, pulmonary, and peripheral vessels. PMID- 9352177 TI - The impact of lipid lowering in coronary artery disease. AB - A growing number of clinical trials using various methods of lowering cholesterol levels have demonstrated that cholesterol lowering decreases the risk of coronary disease in both primary and secondary prevention. Recent clinical and angiographic trials have shown beneficial effects of cholesterol lowering in patients with average cholesterol levels and with other lipid abnormalities. Studies using aggressive low-density lipoprotein (LDL) lowering have demonstrated improvements in angiographic endpoints and the functional capacity of patients. Data from other studies suggest improvement in endothelial function and in ischemia with LDL lowering. The challenge for the future is to extend the benefits of therapy to more patients and to expand our knowledge about other lipid and nonlipid risk factors to decrease the risk of coronary heart disease. PMID- 9352180 TI - Clinical trials. PMID- 9352181 TI - Ischemic heart disease. PMID- 9352182 TI - Recombination in the mammalian germ line. AB - Elucidation of meiotic recombination mechanisms in mammals faces many obstacles. Much of our understanding has been built upon studies in the fungi, which have served to guide experimental design in mammalian cells and mice. A clearer picture is now emerging which reveals that many of the general principles of recombination are conserved across this evolutionary divide. A number of genes critical to meiotic recombination in yeast also exist in mammals. Transgenic technologies, in addition to advances in molecular biology, now provide several strategies to investigate the properties and regulation of mammalian recombination. This chapter reviews the current state of knowledge regarding recombination in the mammalian germ line, covering topics such as gene conversion, recombination mechanics, recombination-based genetic mutation, crossing over, and genes involved in meiotic recombination. PMID- 9352184 TI - Pairing sites and the role of chromosome pairing in meiosis and spermatogenesis in male Drosophila. AB - Mechanistic and regulatory aspects of meiotic chromosome pairing and segregation have received increasing attention in recent years. This review is concerned with the role of chromosomal sites and chromosome organization in pairing and sperm development in Drosophila. Two major topics are reviewed. The first concerns the distribution and identification of meiotic pairing sites in male Drosophila. Cytogenetic data show that pairing sites are distributed widely in the euchromatin of autosomes but are absent from centromeric heterochromatin. The reverse distribution holds for the X, where the major pairing site is located in the central region of the centric heterochromatin, co-mapping with the rDNA locus. Recent transgenic studies have demonstrated that this pairing site consists mainly of a 240-bp repeated sequence in the intergenic spacers of the rDNA repeats. These spacer repeats contain RNA polymerase I promoters, which must be functional for the repeats to have pairing activity, suggesting a mechanistic connection between pairing and transcription. The general idea that pairing sites coincide with transcribed sequences is discussed. The second major topic involves the effects of sex chromosome rearrangements on spermiogenesis. A variety of rearrangements involving the sex chromosomes, including heterochromatic deletions and translocations with autosomes, have been shown to lead either to meiotic drive or to sterility. Recent evidence strongly implicates the X chromosome pairing site in the etiology of these effects. These findings are discussed in terms of a novel model that interprets the spermiogenic disruptions associated with sex chromosome rearrangements as resulting from disabling of spermatids due to triggering of a checkpoint concerned with monitoring chromosome alignment at meiotic metaphase. PMID- 9352185 TI - Functions of DNA repair genes during meiosis. AB - One of the most basic functions in any organism is DNA repair. In addition, programmed DNA "damage," in the form of DNA double-strand breaks (DSBs), is a regular part of the physiology of most organisms. There are three main types of DSB repair: homologous recombination; single-strand annealing; and nonhomologous end joining. The gene products known to be required for these repair processes are conserved in evolution, but the relative dependence on different pathways for DSB repair is different when systems are compared. In the yeast Saccharomyces cerevisiae, the formation and repair of DNA double-strand breaks (DSBs) is apparently an essential feature of meiotic recombination. However, it is not clear whether DSBs are a conserved feature of meiotic recombination in eukaryotes. The basidiomycete Coprinus cinereus presents an experimental system which is amenable to genetic analysis, processes DSBs in a manner similar to complex eukaryotes, and has a naturally synchronous meiosis. An understanding of the functions of conserved genes in DSB repair in C. cinereus and other similar systems will help to determine whether DSB repair is a unifying theme in meiotic recombination or whether conserved gene products have other essential functions that tie together DNA repair and meiosis. PMID- 9352186 TI - Gene expression during mammalian meiosis. AB - The expression of a wide variety of genes is developmentally regulated during mammalian meiosis. Drawing mainly on studies in spermatogenesis, this review shows that some of these genes are transcribed exclusively in germ cells, while others are also transcribed in somatic cells. Some of the genes expressed exclusively in spermatogenic cells are unlike any expressed in somatic cells, while others are isologous to genes expressed in somatic cells and are in the same gene family. Some of the developmentally regulated genes also expressed in somatic cells produce spermatogenic cell-specific transcripts, while others produce transcripts that are apparently the same in somatic and germ cells. Possible answers to why so many genes have atypical patterns of expression during meiosis are that: (1) all cell types express certain genes that define their cell type and lineage, (2) spermatogenesis is a developmental process that progresses according to a genetic program directing the sequential and coordinate expression of specific genes, (3) some genes are expressed that encode proteins required for meiosis, (4) some genes are expressed that encode proteins not required until after meiosis, (5) some genes are expressed to compensate for other genes that become inactivated with X chromosome condensation, and (6) it has been suggested that regulation of gene expression becomes leaky during spermatogenesis due to changes in DNA organization, leading to production of irrelevant transcripts. However, it is largely unknown how extrinsic cues from the endocrine system and surrounding somatic cells interact with intrinsic mechanisms of germ cells to activate signal transduction processes regulating transcription during mammalian meiosis. PMID- 9352187 TI - Caught in the act: deducing meiotic function from protein immunolocalization. AB - Meiotic division comprises a complex series of events, many of which are unique in the life cycle of the organism. The process utilizes both proteins that participate in normal mitotic cell cycle progression and DNA damage repair and proteins expressed only during meiosis. Until recently, few meiotic protein participants had been identified and characterized, but several recent developments have changed this situation. Proteins can be selected for study based on their cDNA sequence and similarity to known proteins with "suspicious" repair/recombination or cell cycle activity and antibodies against these proteins applied to meiotic nuclei to test for activity. With the development of gene sequence data bases from many organisms, similarity to a known protein need not be based on the same or even a closely related species. Potential interactions between two or more proteins can be identified and involvement in a common process inferred based on antibody colocalization. The gene sequence can be disrupted and the effect on meiotic progression directly examined. Previously identified structures, the synaptonemal complex (SC) and both early and late recombination nodules (RNs), provide structural and temporal landmarks that assist in inferring meiotic activity of the protein being studied. Mammalian meiosis is especially attractive for these kinds of studies since spermatocyte and oocyte nuclei are large with distinct nuclear organelles and since meiosis is highly protracted, occurring over a period of several days. In this chapter, an approach to the study of mammalian meiosis based on use of specific antibodies is outlined and methods of coupling this approach to other techniques, such as targeted gene disruption or chromosome aberrations, are described. Some of the proteins already identified as participants in meiotic prophase are reviewed and their presumed functions discussed. PMID- 9352188 TI - Chromosome cores and chromatin at meiotic prophase. AB - We review the synaptonemal complex, SC, of the synapsed homologous chromosomes at meiotic prophase in insects and mammals in terms of its formation, and the association of specific chromatin elements with the synaptonemal complexes. The focus is: (1) The SC as visualized with a variety of techniques; (2) The nature of the chromatin loops where they are associated with the SCs--the bases of the loops may be instrumental in recombinant events judging from the presence of Rad51 protein and late recombination nodules at the SCs; (3) Differences in DNA content of similarly sized loops; (4) Requirements for chromatin attachment to the chromosome cores, requirements that are apparently lacking in foreign DNA inserts; (5) Regulation of loop size by the position along the chromosome; (6) The structural correlates of recombination at the SCs--these comments are based on studies of SC structure, DNA-core protein associations, fluorescent in situ hybridization to visualize specific DNA segments, and fluorescent immunocytology to visualize the chromosome core proteins. PMID- 9352189 TI - Chromosome segregation during meiosis: building an unambivalent bivalent. AB - Faithful chromosome segregation during anaphase requires that stable microtubule connections are established between chromosomes and both spindle poles by metaphase. Bipolar orientation follows an active period of transient connections between the kinetochores and poles, and tension mediated through attachments between the chromosomes stabilizes those bivalents that have connections to opposite poles. This review focuses on how the chromatids are tied together in the bivalent to ensure proper segregation in the two meiotic divisions. Homologs are partitioned in meiosis I, and reciprocal crossovers, cytologically defined as chiasmata, usually hold the homologs together for this division. The crossovers themselves must be prevented from migrating off the chromatid arms. Binding substances localized to the crossover and sister-chromatid cohesion distal to the crossover have been proposed to prevent loss of chiasmata. Spontaneous nondisjunction events and mutations that disrupt the maintenance of chiasmata are analyzed in the context of these models. Homologs that segregate in meiosis I without chiasmata are briefly discussed. The bivalent must also be constructed so that four chromatids present only two functional kinetochores prior to anaphase I. Cytology and genetic data suggest that the sister kinetochores are duplicated but constrained to act as a single kinetochore. Additionally, centromeric regions of sister chromatids preserve their cohesion until anaphase II, even as cohesion on the sister-chromatid arms is lost at anaphase I. Mutations that specifically disrupt this process are presented. PMID- 9352183 TI - Meiotic recombination hotspots: shaping the genome and insights into hypervariable minisatellite DNA change. AB - Meiotic homologous recombination serves three principal roles. First, recombination reassorts the linkages between newly-arising alleles to provide genetic diversity upon which natural selection can act. Second, recombination is used to repair certain types of DNA damage to provide a mechanism of genomic homeostasis. Third, with few exceptions homologous recombination is required for the appropriate segregation of homologous chromosomes during meiosis. Recombination rates are elevated near DNA sites called "recombination hotspots." These sites influence the distribution of recombination along chromosomes and the timing of recombination during the life cycle. Recent advances have revealed biochemical steps of hotspot activation and have suggested that hotspots may regulate when and where recombination occurs. Two models for hotspot activation, one in which hotspots act early in the recombination pathway and one in which hotspots act late in the recombination pathway, are presented. The latter model can account for changes at hypervariable minisatellite DNA in metazoan genomes by invoking resolution of Holliday junctions at minisatellite DNA repeats. PMID- 9352190 TI - Regulation and execution of meiosis in Drosophila males. AB - In this chapter we review the regulation and execution of the meiotic cell divisions in the context of the developmental program that comprises Drosophila spermatogenesis. Male germ line cells undergoing meiosis are readily identifiable and are of a size and abundance that makes this system well suited for morphological characterizations of cell division. Furthermore, a wide range of molecular genetic techniques are available, facilitating mechanistic investigations. We present an overview of key stages in spermatogenesis and, in particular, meiosis. We consider the pathways controlling entry into the meiotic divisions in the context of established cell cycle regulators as well as newly identified loci required for meiotic entry. We then review the assembly and function of both the meiotic spindle and the contractile ring. We conclude with a consideration of questions and problems that await further investigation. PMID- 9352191 TI - Sexual dimorphism in the regulation of mammalian meiosis. PMID- 9352192 TI - Genetic control of mammalian female meiosis. PMID- 9352193 TI - Nondisjunction in the human male. PMID- 9352194 TI - A major glycoprotein of Xenopus egg vitelline envelope, gp41, is a frog homolog of mammalian ZP3. AB - A predominant glycoprotein in the vitelline envelope (VE) of the anuran Xenopus laevis is gp41, known to be proteolytically converted from gp43 of the coelomic egg envelope concomitant with the acquisition of egg fertilizability. To characterize the protein core of gp41, purified gp41 from VE was digested with lysyl endopeptidase, and peptides isolated from the digests were sequenced for amino acids to design degenerate primers for polymerase chain reaction. By reverse transcription-polymerase chain reaction with a poly(A)+ RNA from the ovary of an ovulated female Xenopus, a specifically amplified band was obtained and sequenced. The upstream and downstream sequences of the sequenced region were completed by 5'- and 3'-rapid amplification of cDNA ends, respectively. The cDNA, referred to as gp43 cDNA, comprises 1423 base pairs and contains one open reading frame with a sequence for 460 amino acids. The predicted amino acid sequence of gp43 cDNA has a close similarity with that of mammalian ZP3. Northern blot and in situ hybridization studies indicated that gp43 mRNA is expressed in oocytes, particularly in the previtellogenic oocytes. A comparison of the N-terminal sequences of gp41 and gp43 strongly suggested that gp41 is generated at least by processing of the N-terminal portion of gp43 with oviductin. PMID- 9352195 TI - Probable participation of phospholipase A2 reaction in the process of fertilization-induced activation of sea urchin eggs. AB - In sea urchin eggs activated by sperm, A23187 or melittin, BPB (4-bromophenacyl bromide, a phospholipase A2 inhibitor) blocked fertilization envelope formation and transient CN(-)-insensitive respiration in a concentration-dependent manner. BPB had virtually no effect on the increase in [Ca2+]i (cytosolic Ca2+ level), the activity of phosphorylase a and the rate of protein synthesis, as well as acid production and augmentation of CN(-)-sensitive respiration. BPB also inhibited fertilization envelope formation and augmentation of CN(-)-insensitive respiration induced by melittin. Melittin, known to be an activator of phospholipase A2, induced the envelope formation, acid production, augmentation of CN(-)-insensitive and sensitive respiration, but did not cause any increase in [Ca2+]i, the phosphorylase a activity and the rate of protein synthesis. An activation of phospholipase A2 induced by Ca2+ or melittin seems to result in cortical vesicle discharge and production of fatty acids, which are to be utilized in CN(-)-insensitive lipid peroxidase reactions. Activation of other examined cell functions in eggs activated by sperm or A23187, probably results from Ca(2+)-triggered sequential reactions other than Ca(2+)-caused activation of phospholipase A2. PMID- 9352196 TI - Cell ablation by ectopic expression of cell death genes, ced-3 and Ice, in Drosophila. AB - We have developed a system for killing specific cells in Drosophila using ectopic expression of cell death genes. CED-3 and ICE (caspase-1) are proteins required for programmed cell death in the nematode Caenorhabditis elegans and in mammals, respectively. Our previous study has shown that both ced-3 and Ice can elicit cell death in Drosophila. By expressing ced-3 or Ice in several kinds of cells using a GAL4-UAS system and examining the resulting morphological defects, we show that these abnormalities are thought to be caused by the action of ced-3 or Ice genes. As cells are killed by apoptosis in our system, we could eliminate the possibility of harmful effects on the neighboring cells. Our system provides an alternative and novel cell ablation method to elucidate mechanisms of cell differentiation and cell-cell interactions during development in Drosophila. PMID- 9352197 TI - Specific cellular localization of tyrosinase mRNA during Ciona intestinalis larval development. AB - A Ciona intestinalis cDNA clone that encodes a protein highly homologous to other tyrosinases was isolated. Northern blot analysis showed that expression of Ciona tyrosinase starts at the early neurula stage and continues throughout the tail bud and tadpole larval stages. The earliest tyrosinase expression was detected, by in situ hybridization, at the neural plate stage, in pigment precursor cells located along the two neural folds, in the animal region of the embryo. In the course of embryonic development the strong hybridization signal was always localized, within the rostral part of the developing brain, in the pigment precursor cells and was later detected in the otolith and ocellus. These results are discussed in relation to tyrosinase as an early marker of neural induction. PMID- 9352198 TI - Bh (black at hatch) gene appears to be expressed in melanocytes of feather germs in the Japanese quail (Coturnix coturnix japonica). AB - The Bh (black at hatch) gene was examined to determine whether it is expressed in plumage melanocytes by analyzing pigmentation patterns of Bh melanocytes placed in the micro-environment of the feather germs of quail embryos with pink eyes. These host quails genetically lack a large part of plumage melanin. The Bh locus in these almost white quails is wild-type. When Bh neural crest cells were transplanted orthotopically into the host embryos, wild-type and Bh/+ melanocytes, which differentiated from the transplanted neural crest cells, formed plumage pigmentation patterns characteristic of each genotype in the micro environment of the host feather germs. Brown plumage pigmentation, which was very similar to that of 10-day Bh/Bh embryos, was also observed in the feather germs of host embryos that received Bh neural crest cells, although the genotype of the donors could not be determined. These donors died before pigmentation of their feather germs occurred. The results demonstrate that pigmentation patterns of Bh melanocytes are not altered in the micro-environment of the host germs, suggesting that the Bh gene is autonomous in Bh melanocytes and is expressed in melanocytes of both Bh and the host feather germs, and that it causes the normal pigmentation pattern to be altered. PMID- 9352199 TI - N-cadherin is crucial for heart formation in the chick embryo. AB - The developing heart primordium strongly expresses N-cadherin. In order to investigate the role of this adhesion molecule in heart morphogenesis, chicken embryos were cultured at stages 5-12, and injected with anti-N-cadherin antibodies that can specifically block the activity of this cadherin. In the injected embryos, the epimyocardial layers, which develop bilaterally from the splanchnic mesoderm, did not fuse to form a single cardiac tube. Moreover, each of the unfused layers became fragmented into epithelioid clusters. At the cellular level, large intercellular gaps were observed in the antibody-treated myocardial layers. These disorganized myocardial layers beat to some extent, suggesting that their differentiation was not blocked; however, their contraction was not coordinated. Morphogenesis of other tissues, not only N-cadherin-negative but also N-cadherin-positive tissues, such as the neural tube and notochord, proceeded normally even in the presence of anti-N-cadherin antibodies. These results suggest that N-cadherin is indispensable for heart formation, but not for morphogenesis of the other tissues, at the developmental stages examined. For the latter processes, expression of other cadherin subtypes presumably compensated for the loss of N-cadherin activity. PMID- 9352200 TI - cDNA cloning and sequence analysis of the Xenopus laevis egg envelope glycoprotein gp43. AB - The glycoproteins of the Xenopus laevis egg envelope function in fertilization and development. As the unfertilizable coelomic egg transits the pars recta region of the oviduct, it is converted to a fertilizable egg by limited proteolysis of the envelope glycoprotein gp43 to gp41. This conversion is caused by an oviductally secreted serine active site protease, oviductin. We cloned a cDNA for gp43 from an oocyte cDNA library. The cDNA encoded a 454 amino acid protein homologous to the ZPC family of glycoproteins previously shown to be present in mammalian and fish egg envelopes. Conserved ZPC domains and motifs present in the Xenopus sequence included a signal peptide sequence, an N-linked glycosylation site, and 12 aligned Cys residues. In mammalian and Xenopus sequences, a furin-like (convertase) site and a C-terminal transmembrane domain were present reflecting the biosynthesis of ZPC in these species via the secretory glycoprotein pathway. However, fish envelope glycoproteins lack these sequences since they are synthesized via a different route (in the liver, transported to the ovary, and assembled into the egg envelope surrounding the oocyte). Consensus amino acid residues were identified by sequence comparisons of seven ZPC family members; 19% of the amino acid residues were invariant and 48% of the residues were identical in at least four of the seven sequences. The consensus sequence was used to make structure-fertilization function predictions for this phylogenetically conserved family of glycoproteins. PMID- 9352202 TI - Conditions for a heat shock response during oogenesis and embryogenesis of the amphibian Pleurodeles waltl. AB - The optimal conditions capable of inducing an increase in HSP70 neosynthesis during development of the urodele amphibian Pleurodeles waltl were determined in this study. These conditions depend on temperature, heat shock duration and recovery duration. In oocytes, a heat shock response was repeatedly obtained at 37 degrees C for 15 min followed by 1 h recovery. These results provided evidence for heat shock response at every stage considered. An increase in HSP70 synthesis was noted throughout oogenesis, but it did not lead to an increase in the amount of soluble HSP70, except for stage VI oocytes. Such results suggest that from stage II to stage IV oocytes, an equilibrium occurs between the HSP70 used and the HSP70 neosynthesized. In contrast, in stage VI oocytes, heat shock led to overproduction of HSP70. During early development, the heat shock response was repeatedly obtained only from the gastrula stage with a 37 degrees C shock and a 15 min duration of treatment. Surprisingly, during cleavage stage, the soluble HSP70 total amount increased after heat shock at a time when no HSP70 neosynthesis occurred. PMID- 9352203 TI - Color pattern formation on the wing of a butterfly Pieris rapae. 2. Color determination and scale development. AB - It has been shown that microcautery on the prospective apical black region of the early pupal forewing of a butterfly, Pieris rapae, causes alteration of the scale color on the adult wing and a delay in histogenesis of the pupal wing. From these results, it has been assumed that the developmental delay of scale cells in the pupal wing alters their developmental fate and the hypothesis that different color fates of scales are determined by differences in the developmental timetables between scale cells is proposed. In this study, we attempted to find the developmental timetables of individual scales expressing specific color to test this hypothesis. It was found that the holes on the upper surface of a scale become larger as they develop and the hole sizes of scales in the white region are always larger than in the black region on the same wings either during pupal period or after eclosion. This suggests that the scale hole size is a good index that reflects developmental rate of the scale and a difference in the hole size between adult scales is attributed to a difference in the developmental timetables when their ancestral scale precursor cells were in the pupal period. A comparison of the hole sizes between adult scales in different color regions suggested that normal white scales were in a more advanced state than were the black ones but white scales induced by microcautery were in a less advanced state than black ones on the same wing. This supports our hypothesis. PMID- 9352201 TI - A cis-regulatory element within the 5' flanking region of arylsulfatase gene of sea urchin, Hemicentrotus pulcherrimus. AB - The 5' flanking region of the sea urchin Hemicentrotus pulcherrimus arylsulfatase (Ars) gene was scanned to define cis-regulatory elements required for proper expression congruent to that of the endogenous gene. The region between -100 bp and +38 bp from the transcription start site contains minimum information required for temporal initiation of transcription of the Ars gene. Progressive deletion analysis of Ars-luciferase reporter constructs containing the Ars sequence from -3484 bp to +38 bp suggests the existence of several cis-regulatory elements within this region. Results from luciferase assays of internal deletion mutants show strong enhancer activity detected within the sequence from -194 bp to -144 bp. By gel mobility shift assay, we have identified a nuclear factor that interacts sequence-specifically with this 50 bp region, and appears in a developmental stage-specific manner. Further deletion analysis determined that the enhancer activity lies within a 22 bp sequence between -186 bp and -164 bp. PMID- 9352204 TI - Murine forebrain and midbrain crest cells generate different characteristic derivatives in vitro. AB - Neural crest (NC) is a transient structure that gives rise to various types of tissues. Many NC cells are pluripotent in the sense that their progeny can generate more than one derivative. However, the potentiality to differentiate into certain derivatives, such as cartilage and bone, seems to be specified with respect to the neuraxial levels at which the NC generates. In order to compare the differentiation potentiality of different regions of head NC, the derivatives of forebrain and midbrain mouse NC have been investigated in vitro using explant cultures of neuroepithelial fragments. From morphology and expression of specific markers, the midbrain crest cultures obviously generated earlier and were greater in number of neuronal cells than were the forebrain ones. Moreover, collagen type II positive cells were detected in the midbrain but not in the forebrain crest cultures. Finally, pigment cells were only observed in the forebrain cultures. The results suggest that the forebrain and midbrain crest cells have a different potentiality to differentiate. PMID- 9352205 TI - Adrenergic neurotransmitters and calcium ionophore-induced situs inversus viscerum in Xenopus laevis embryos. AB - Xenopus laevis embryos at the blastula-early tail bud stage were exposed to norepinephrine or octopamine dissolved in culture saline until they reached the larval stage. The left-right asymmetry of the heart and gut was then examined. We found that these adrenergic neurotransmitters induced situs inversus in the heart and/or gut in up to 35% of tested neurula embryos. Norepinephrine-induced situs inversus was blocked by the alpha-1 adrenergic antagonist prazosin. Furthermore, A23187, a calcium ionophore, also increased the incidence of situs inversus up to 54% when late-neurula embryos were exposed to the solution. A23187 treatment initiated before neural groove formation was less effective. The incidence of situs inversus induced by these reagents decreased towards the control level (2.2%, 25 untreated embryos out of 1127 embryos in total) in embryos past the stage of neural tube closure. In the present experiments we obtained 22 gut-only situs inversus embryos having an inverted gut and a normal heart. In contrast, such embryos were not observed among the 1127 untreated embryos. An adrenergic signal mediated by an increase in intracellular free calcium may be involved in the asymmetrical visceral morphogenesis of Xenopus embryos. PMID- 9352206 TI - Double-segment defining role of even-skipped homologs along the evolution of insect pattern formation. AB - Recent studies on insect patterning suggest that the genetic hierarchy may be roughly conserved in phylogenetically divergent species, but pair-rule genes may not function identically in all insects. In order to understand potential evolutionary changes in the role of the pair-rule genes, a Bombyx even-skipped homolog was cloned and its expression pattern during early embryogenesis studied. Eight stripes of Bombyx even-skipped were progressively expressed in an antero posterior order. Later, these stripes disappeared anteriorly. Under this detection system, Bombyx even-skipped stripes clearly do not resolve into the corresponding secondary stripes, an obvious difference from Drosophila and Tribolium. These results suggest that Bombyx even-skipped may serve a double segment defining role and may determine the odd-numbered engrailed stripes. PMID- 9352207 TI - Activation of protein kinase C induces cortical granule exocytosis in a Ca(2+) independent manner, but not the resumption of cell cycle in porcine eggs. AB - The effects of protein kinase C (PKC) activation on meiotic resumption and cortical granule (CG) exocytosis as well as its dependence on Ca2+ in porcine eggs matured in vitro were studied. Cortical granule release was judged by both confocal laser microscopy after the eggs were labeled with fluorescein isothiocyanate-peanut agglutinin (FITC-PNA) and electron microscopy. Meiotic resumption and pronuclear formation were observed after eggs were stained with acetic orcein. When eggs were treated with PKC activators, 1-oleyl-2-acetyl glycerol (OAG) or phorbol 12-myristate 13-acetate (PMA), the pronuclear formation percentage was significantly lower than that of Ca2+ ionophore A23187-treated group, but not statistically different from that in negative control group (P > 0.05), and most of the eggs were still arrested at metaphase II stage, suggesting that PKC activation does not induce the resumption of meiosis and pronuclear formation. In contrast, PKC activation induced 89.1% to 100% of the eggs completely or partially released their CG in different groups, not statistically different from A23187-treated group, and this effect could be overcome by PKC inhibition. When the intracellular free Ca2+ was chelated with acetoxymethal ester form of 1,2-bis(O-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA AM), and then treated with PMA or OAG in Ca(2+)-free medium, the proportions of eggs with CG release were 90.9% and 78.1%, respectively, not statistically different from the above-treated groups, suggesting that CG exocytosis induced by PKC activation is independent of Ca2+ rise. The results indicate that different events of porcine egg activation may be uncoupled from one another. PMID- 9352208 TI - An embryological study of ventralization of dorsal structures in the tail of medaka (Oryzias latipes) Da mutants. AB - In adult Da (double anal fin) mutants of medaka (Oryzias latipes), structures such as the dorsal fin and the dorsal half of the caudal fin are ventralized in adult fish. However, there have been few embryological studies of the development of mutant phenotypes except those of the caudal fin. In this study, development of mutant phenotypes of the tail where they typically develop was examined morphologically at various stages of embryogenesis. The arrangement of melanocytes along the dorsal midline, the shape of the dorsal fin fold, and the shape of the dorsal myotome exhibited a ventral pattern in the tail at various embryonic stages in Da mutants. PMID- 9352210 TI - Sensitivity of macrophyte-dominated freshwater microcosms to chronic levels of the herbicide linuron. I. Primary producers. AB - Effects of chronic concentrations of linuron (0, 0.5, 5, 15, 50, and 150 micrograms/L) were studied in indoor, macrophyte dominated, freshwater microcosms. The concentrations were kept at a constant level for 4 weeks. This paper is the first in a series of two and summarizes the course of the linuron concentrations in time and its effects on macrophytes, periphyton, and phytoplankton. These endpoints were studied from 3 weeks before the start of the treatment until 11 weeks after the start. The degradation of linuron in the water was lower at higher treatment levels, probably due to a decrease in pH. Linuron treatment resulted in a decrease in biomass of the macrophyte Elodea nuttallii and a clear decrease in abundance of the algae Cocconeis, Chroomonas, and Phormidium foveolarum. It was found that Cocconeis first decreased in biovolume and after 2 weeks also in abundance. The alga Chlamydomonas increased in abundance at the two highest doses, resulting in higher chlorophyll-a levels. The NOECs of 0.5 micrograms/L for the inhibition of the growth and photosynthesis of Elodea nuttallii, the abundance of Cocconeis and Chroomonas, and the oxygen and pH levels were the lowest recorded in the microcosms. The safety factors adopted by the EU in the Uniform Principles appeared to ensure adequate protection for the ecosystem in the case of chronic exposure to linuron. PMID- 9352209 TI - Use of the Vibrio harveyi toxicity test for evaluating mixture interactions of nitrobenzene and dinitrobenzene. AB - A mixture toxicity investigation was conducted using the bioluminescent marine bacterium Vibrio harveyi as the test organism for dual combinations of nitrobenzene and dinitrobenzene. Change in bioluminescence was used for determination of toxicity. Combination toxicity was evaluated using statistical comparisons, isopleths (isobologram and isobole plot), an additive index, and a mixture toxicity index. Both isopleths and mixture toxicity indices suggest that various combinations are additive, while the additive index value suggests antagonism. All evaluations were conducted as equitoxic mixtures. Statistical determination was performed using the z test. Numerous comparisons were different at the 1% level. Slope of line associated with isobole plot was suggested to be an important factor, resulting in statistical differences among comparisons. Distribution, using the Shapiro-Wilk test, was determined for both individual combination groups and solution composition in isopleths. All distributions evaluated were normal. These results suggest that the V. harveyi toxicity test is useful for mixture toxicity studies. PMID- 9352211 TI - Sensitivity of macrophyte-dominated freshwater microcosms to chronic levels of the herbicide linuron. II. Community metabolism and invertebrates. AB - Effects of a chronic application of the herbicide Afalon (active ingredient linuron) on physicochemical conditions, decomposition of plant litter, and densities of zooplankton and macroinvertebrates were studied in indoor microcosms intended to model drainage ditches. For 28 days, concentrations of 0, 0.5, 5, 15, 50, and 150 micrograms/L linuron were maintained, each in two replicates. The microcosms were dominated by the macrophyte Elodea nuttallii. The functional response of the ecosystem is discussed in relation to shifts in community structure. Treatment effects of linuron on community metabolism, as a direct effect of the inhibition of the photosynthesis of macrophytes and algae, resulted in a decrease in dissolved oxygen and pH, and an increase in alkalinity and conductivity (NOEC 0.5 microgram/L). During the posttreatment period, differences between controls and highest dose fell gradually, but were still significant 7 weeks after the start of linuron application. Decomposition of particulate organic material in litter bags was not affected, despite decreases in DO. The negative effect of linuron on several algae (cryptophytes, diatoms) and the positive effect on the green alga Chlamydomonas resulted in a decrease of several Rotatoria and an increase in Copepoda, and, to a lesser extent, Cladocera. The complete disappearance of the macrophyte E. nuttallii in the 150 micrograms/L microcosms and a 50% reduction of its biomass in the 50 micrograms/L microcosms reduced the numbers of the snail Physella acuta, which normally inhabits macrophytes. Artificial substrates indicated a significant increase in the isopod Asellus aquaticus in the 50 and 150 micrograms/L microcosms during the post treatment period. This, however, was counteracted by a significant decrease in A. aquaticus at the final harvest. Changes in the ecosystem structure (decline in macrophyte biomass) made the artificial substrates more attractive. PMID- 9352212 TI - Effect of low-molecular-weight alkanes on the plant cell photosynthetic apparatus. AB - The effects of aliphatic hydrocarbons--methane, ethane, propane, butane, and their mixture--on the photosynthetic apparatus of maize (Zea mays) and raygrass (Arrhenetherum elatius) leaves have been studied. The pathology of subcellular organelles as well as of the whole architectonics of the cell was observed. An especially destructive action of alkanes is expressed on the granalamellae system of chloroplasts. This action is more profound in the upper part of the leaf. PMID- 9352213 TI - Serum "B" esterases as a nondestructive biomarker for monitoring the exposure of reptiles to organophosphorus insecticides. AB - A field study was conducted to validate serum B esterases as nondestructive biomarkers (NDBs) in lizards. Serum butyrylcholinesterase (BChE) and carboxylesterase (CbE) activities were measured in lizards and four species of birds collected in an area of 0.5 ha sprayed with 0.36 kg a.i./ha of Folidol SE5 (5% parathion). Serum B esterase activities were determined in a total of 213 lizards (Gallotia galloti) and 81 birds of four species (Sylvia melanocephala, Serinus canaria, Parus caeruleus, and Erithacus rubecula) collected for 23 days after the spraying. A control group of 39 lizards and 58 birds was sampled before the spraying. No relationship was found between serum B esterases and sex or biometric parameters in all species. Inhibition of BChE (> 40%) and CbE (> 50%) activities was recorded in lizards 23 days after spraying. BChE activity was found to be more sensitive than CbE to inhibition by parathion. Inhibition of serum B esterase activities was recorded in only two bird species (S. melanocephala and S. canaria), but the number of individuals collected was much less than the lizards. The advantages and disadvantages of G. galloti as bioindicator of exposure to organophosphorus insecticides in the Canary Islands (Spain) are discussed in relation to birds commonly used for this purpose. PMID- 9352214 TI - Bioaccumulation of methyl parathion and its toxicology in several species of the freshwater community in Ignacio Ramirez dam in Mexico. AB - Environmental contamination by pesticides, including the presence of chemical residues in aquatic wildlife, is a widespread ecological problem. Methyl parathion (MP), a widely used organophosphorate insecticide, is a potent neurotoxic in both vertebrates and invertebrates. The effect of a subchronic exposure to MP in aquatic organisms was evaluated in a natural ecosystem measuring acetyl cholinesterase (AChE) and gamma glutamil transpeptidase (GGT) activity. Two samples were conducted. Physicochemical characterization was done at each sampling time and organisms were collected. MP and metabolite 4 nitrophenol (4-NP) concentrations were measured in water sediment and organisms. The major differences in physical features between season were an increase of turbidity and salinity and depletion of dissolved oxygen in the rainy season. MP and 4-NP are bioconcentrated in organisms in response to environmental stress. MP concentration was measured in different size/age and reproductive stages separately. A significant concentration in reproductive tissues (plants)/unborn progeny (animals) was always found, and this can affect egg viability. The metabolite 4-NP is bioaccumulated and is toxic because it causes an increase of AChE activity. GGT activity was higher than that in controls. The increase in enzymatic activity provides a detoxification mechanism from chronic sublethal exposure, when hepatic glutation depletion occurs, and may be an indicator of liver damage. PMID- 9352215 TI - Infectivity and effects of gypsy moth and spruce budworm nuclear polyhedrosis viruses ingested by rainbow trout. AB - Rainbow trout fingerlings were fed dried krill injected with gypsy moth or spruce budworm nuclear polyhedrosis virus (LdNPV and CfNPV, respectively) at a total dose of 1.4 x 10(7) occlusion bodies (OBs) per fish. By the end of the 21-day experimental period there were no adverse effects on fish survival or behavior and no significant differences in feeding rates or growth between treated and control fish. The internal organs of all fish were examined at the end of the experiment and there were no signs of lesions, discoloration, swelling, hemorrhaging, or other aberrations. Visceral tissues were analyzed with a horseradish peroxidase-labeled whole genomic DNA probe (enhanced chemiluminescence procedure) to detect infection by the NPVs. There were no indications of NPV infection (no positive signals) in stomach and intestinal tract tissues of treated fish. High background signals were obtained from liver samples, but further analyses indicated that these were not due to the presence of LdNPV or CfNPV. The protocols outlined here should be applicable to determining infectivity and effects of genetically modified insect viruses on fish. PMID- 9352216 TI - Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. AB - Four members of the tissue inhibitor of metalloproteinases (TIMP) family have been characterized so far, designated as TIMP-1, TIMP-2, TIMP-3, and TIMP-4. TIMP 1 and TIMP-2 are capable of inhibiting the activities of all known matrix metalloproteinases (MMPs) and as such play a key role in maintaining the balance between extracellular matrix (ECM) deposition and degradation in different physiological processes. Accelerated breakdown of ECM occurs in various pathological processes, including inflammation, chronic degenerative diseases and tumor invasion. TIMP-1 and TIMP-2 can inhibit tumor growth, invasion, and metastasis in experimental models which has been associated with their MMP inhibitory activity. Recent developments in TIMP research suggest that TIMP-1 and TIMP-2 are multifunctional proteins with diverse actions. Both inhibitors exhibit growth factor-like activity and can inhibit angiogenesis. Structure-function studies have separated the MMP inhibitory activity of TIMP-1 from its growth promoting effect. TIMP-1 has also been implicated in gonadal steroidogenesis and as a cellular elongation factor. TIMP-3 is the only member of the TIMP family which is found exclusively in the extracellular matrix (ECM). It is regulated in a cell cycle-dependent fashion in certain cell types and may serve as a marker for terminal differentiation. The most recently discovered TIMP, TIMP-4, may function in a tissue-specific fashion in extracellular matrix hemostasis. The main aim of this article is to review recent literature on TIMPs with special emphasis on their biological activities and the possibility that they may have paradoxical roles in tumor progression. PMID- 9352217 TI - Intracellular localization and nucleocytoplasmic transport of Ro RNP components. AB - The Ro and La autoantigens, which were originally identified by sera from autoimmune patients, consist of RNA-protein complexes (RNPs), in which the protein components carry most of the autoantigenic determinants. In human cells, Ro RNPs consist of one of four small RNA molecules, termed hY1, hY3, hY4 and hY5 [35], associated with several proteins, Ro60, Ro52 and La [8, 35, 106] and possibly other, yet unidentified polypeptides. Controversial data have been published on the association of the protein calreticulin with Ro RNPs [53, 58, 75, 78], but recently in vitro evidence has been obtained that non-phosphorylated calreticulin is able to interact with hY RNAs directly [17]. The physiological significance of this interaction remains to be established. In addition to its association with hY RNAs, La is (in most cases transiently) associated with all other RNA polymerase III transcripts. The hY RNAs are the only known cellular RNAs stably bound by La. While the Ro RNPs were found to reside mainly in the cytoplasm, the other La RNPs are mainly nuclear. In this review recent progress made on the intracellular localization and transport of Ro RNP components and of assembled Ro RNPs will be discussed. PMID- 9352218 TI - Targeting of green fluorescent protein to neuroendocrine secretory granules: a new tool for real time studies of regulated protein secretion. AB - Human chromogranin B (hCgB), a soluble marker protein of neuroendocrine secretory granules, was fused to green fluorescent protein (GFP). Two GFP-mutants with different folding properties, S65T and EGFP, were used to produce two recombinant proteins, hCgB-GFP(S65T) and hCgB-EGFP, respectively. After transient expression only hCgB-EGFP elicited green fluorescence in the neuroendocrine cell line PC12. Pulse-chase experiments with [35S]sulfate followed by subcellular fractionation showed that hCgB-EGFP was sorted with high efficiency to immature secretory granules (ISG). Confocal microscopy revealed that fluorescent hCgB-EGFP colocalized largely with synaptotagmin, a membrane marker of secretory granules and synaptic-like microvesicles, and significantly with endogenous rat chromogranin B (rCgB), a soluble marker of secretory granules. Upon stimulation of transfected cells with 5 mM Ba2+ or by depolarization with 50 mM K+ hCgB-EGFP underwent regulated exocytosis. The dynamics of green fluorescent secretory granules beneath the plasma membrane (PM) of living PC12 cells were visualized by confocal microscopy. The majority of these vesicles did not move within 8.5 sec as if they were docked. In contrast, in NGF-induced cells most of the secretory granules beneath the somatic PM moved within the same time period whereas only little movement was observed in the neurites. These findings indicate that in differentiated PC12 cells the majority of the docking zones are not in the soma but are distributed along the neurites. In conclusion, the fusion protein hCgB EGFP provides a powerful tool to study in real time vesicular traffic in the regulated pathway of protein secretion. PMID- 9352219 TI - Rat homologues of yeast sec7p. AB - Mutations in the Saccharomyces cerevisiae sec7 locus lead to a pleiotropic secretory phenotype that is characterized by an accumulation of Golgi cisternae and a loss of secretory granules. This indicates that the corresponding gene product sec7p is involved in the budding of secretory granules from the Golgi apparatus. Here we report the primary structure of three rat homologues of sec7p, called msec7-1, -2, and -3. The mRNAs of these genes are expressed in all tissues tested. All msec7s share the same domain structure in which an N-terminal coiled coil domain is followed by a sec7-homology domain and a pleckstrin-homology domain. On the protein level, msec7s are present in all rat tissues tested, with highest protein levels in brain and adrenal. In the adult rat brain, they are present in soluble and membrane-associated pools. PMID- 9352220 TI - Protein segregation in peripheral 15 degrees C intermediates in response to caffeine treatment. AB - Previous studies have shown that caffeine treatment at 20 degrees C causes the intermediate compartment protein p58 to redistribute from the Golgi region without affecting the localization of the Golgi stack protein mannosidase II (J. Jantti, E. Kuismanen, J. Cell Biol. 120, 1321-1335 (1993). Here we have dissected further the effect of caffeine on transport of Golgi and intermediate compartment proteins from the cell periphery to the perinuclear Golgi region. To accumulate proteins in the peripheral membranes, BHK-21 cells were treated with brefeldin A to redistribute marker proteins towards the ER. Following BFA wash-out and subsequent incubation at 15 degrees C, p58, the coat protein beta-COP, and Man II were all localized in the peripheral 15 degrees C-intermediates. When the cells were shifted from 15 degrees C to 20 degrees C all the proteins were recentralized to the Golgi region. However, if the temperature shift was carried out in the presence of 10 mM caffeine, p58 and beta-COP maintained their peripheral localization, whereas Man II was transported to the Golgi region. The results indicate that caffeine at 20 degrees C does not block the centralization of Man II from peripheral sites to the central Golgi region. Therefore, its effect on ER to Golgi transport appears to be manifested specifically at ER exit. Furthermore, our results indicate that segregation of intermediate compartment and Golgi stack proteins can occur at the level of the peripheral 15 degrees C intermediates. Immunoelectron microscopic localization of p58 and Man II showed that these peripheral intermediates consisted of tubules and small stacks of cisternae. Within the tubular intermediates both p58 and Man II appeared to segregate to membrane subdomains. Finally, examination of serial and thick sections support the idea that the stacked structures can be generated from tubular intermediates. PMID- 9352221 TI - Altered distribution of plectin/HD1 in dystrophinopathies. AB - Plectin/HD1 is a high molecular weight protein (approximately 500 kDa) that has been proposed to act as an important and versatile cytoskeletal cross-linker molecule. Mutations of the human plectin gene have recently been associated with the autosomal recessive disorder epidermolysis bullosa simplex with muscular dystrophy. We studied the expression of plectin/HD1 in various neuromuscular disorders by indirect immunofluorescence. In cross sections of normal human muscle, plectin/HD1 showed a checkerboard-like distribution with moderate to intense cytoplasmic and sarcolemmal staining in type 1 fibers and a faint staining of the sarcolemma in type 2 fibers. In longitudinal sections of plectin/HD1-positive fibers a cross-striation staining pattern was noted. This fiber type-related expression was significantly altered in the group of dystrophinopathies, whereas it was maintained in all other myopathies and denervating disorders. In seven dystrophinopathies studied, a markedly increased plectin/HD1 immunoreactivity at the sarcolemmal level of type 2 fibers was observed. Confocal laser microscopy of normal skeletal muscle revealed a colocalization of desmin and plectin/HD1 at the level of the sarcolemma. This suggests that plectin/HD1- in analogy to its demonstrated involvement in cytokeratin-hemidesmosome linkage in epidermis-may mediate the anchorage of desmin to the sarcolemma (i.e. to costameres). PMID- 9352222 TI - Increased expression of CD44 on astrocytoma cells induced by binding myelin basic protein. AB - An astrocytoma cell line (HTB-14), expressing high amounts of a CD44 variant compared to other astrocytoma lines was shown to bind myelin basic protein to a greater extent than low expressing lines in a concentration-dependent manner. The CD44 variant expressed by HTB-14 cells was determined to migrate in sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular mass of 100 kDa compared to that from white matter which had a molecular mass of 80 kDa. The most cationic component of myelin basic protein (MBP), (component 1) bound more avidly than the least cationic isomer (component 8). Internalization of MBP was demonstrated by immunogold electron microscopy and was localized to the perinuclear area with some gold particles in the cytoplasm but not near the plasma membrane. Colocalization with glial fibrillary acid protein suggested an interaction between these two molecules. Binding and internalization of MBP was accompanied by an increase in CD44 as determined by quantitation of gold particles and the measurement of CD44 by sandwich enzyme-linked immunosorbent assay. The implication of these studies for the mechanism of demyelination is discussed. PMID- 9352223 TI - Nucleolar evolution and coiled bodies during meiotic prophase in Olea europaea: differential localization of nucleic acids. AB - We studied the ultrastructural evolution of the nucleolus during meiotic prophase in olive microsporocytes. During prophase, nuclear bodies morphologically similar to coiled bodies were observed. The nucleic acid composition of these bodies was examined in microsporocytes using electron microscopic techniques with EDTA preferential ribonucleoprotein staining, anti-DNA immunolabeling, the in situ terminal deoxynucleotidyl transferase-immunogold technique, and in situ hybridization with 18S rRNA and U3 snoRNA digoxigenin-labeled probes. The ultrastructural appearance of the meiocyte nucleolus indicated a low level of activity from the early prophase stage: the granular component was practically absent and nucleoli were constituted almost exclusively by dense fibrillar component containing large fibrillar centers that lacked chromatin inclusions. However, the appearance of reactivation vacuoles in the nucleolus during zygotene and high levels of rRNA in the nucleoplasm during pachytene support the presence of a peak in rRNA synthesis. Our results also show that the nuclear bodies that appear during prophase I are ribonucleoproteinaceous in nature; neither DNA nor ribosomal RNA were detected. The presence of U3 snoRNA, as shown by in situ hybridization in nuclear bodies from plant material, is also evidence that these structures are coiled bodies. We suggest that coiled bodies are involved not only in pre- and post-splicing events but also in the storage, transport or recycling of rRNA maturation elements. PMID- 9352224 TI - Bile acid-induced morphological changes in hepatoma cells with elevated sodium dependent bile acid uptake capacity. AB - McNtcp.24 cells are rat hepatoma cells that were made competent to take up conjugated bile acids actively from the culture medium. Treatment of McNtcp.24 cells with certain species of bile acids caused significant changes in cell structure. Incubation of McNtcp.24 cells in medium containing 100 microM taurocholic acid induced a profound alteration of cellular morphology. Very larger vesicles, visible by phase contrast microscopy, were the most prominent feature of bile acid-treated McNtcp.24 cells. Staining of cells with Oil red O and filipin indicated that the vesicles did not contain neutral lipids or free cholesterol. The vesicles remained in the cells after efflux of radiolabeled taurocholic acid from bile acid loaded cells, indicating that these structures are not intracellular stores of bile acids. Electron microscopic analysis of bile acid-treated McNtcp.24 cells confirmed that the vesicles were localized within the cells. Taurine-conjugated bile acid species were generally potent inducers of the morphological changes, although tauroursodeoxycholic acid did not have a significant effect. Unconjugated bile acid species were ineffective or only mildly effective. Bile acid treatment also caused profound alteration of mitochondrial structure. Surprisingly, there was no significant effect on the ability of treated cells to oxidize fatty acids. The bile acid-treated cells remained viable and upon withdrawal of bile acids from the culture medium, the cells returned to normal morphology by 24 h. The morphological changes observed after treatment of McNtcp.24 with bile acids are reminiscent of the morphological changes observed in hepatocytes following induction of cholestasis. PMID- 9352225 TI - Interaction of lipoproteins with type II pneumocytes in vitro: morphological studies, uptake kinetics and secretion rate of cholesterol. AB - Apart from dipalmitoyl phosphatidylcholine, cholesterol is the most abundant surfactant lipid. About 90 to 99% of cholesterol of the alveolar surfactant is derived from serum lipoproteins. The aim of this study was to identify the lipoprotein which preferentially supplements type II pneumocytes with cholesterol destined for surfactant production. Ultrastructural investigations revealed that type II pneumocytes bind and take up HDL, LDL and VLDL. Binding and uptake of VLDL occurred even in the presence of excess LDL indicating that, besides LDL receptors, type II pneumocytes express additional binding sites for VLDL. Type II pneumocytes in primary culture are able to take up cholesterol added in the form of HDL, LDL and VLDL. Cholesterol uptake was lowest from HDL and highest from VLDL. The maximal velocity of cholesterol uptake from VLDL was more than three times that of cholesterol uptake from LDL. The half-maximal saturation of cholesterol uptake from VLDL was nearly half that of LDL. From these kinetic data and the distribution of free cholesterol among the serum lipoproteins, we calculated that the cholesterol uptake from VLDL is more than three times that of cholesterol uptake from LDL. In double-labeling experiments type II pneumocytes secreted palmitic acid-labeled phospholipids together with labeled free cholesterol taken up from lipoproteins. The secretion rates of both phospholipids and free cholesterol were stimulated to nearly the same extent by isoproterenol. From our results we conclude that type II pneumocytes interact specifically with HDL, LDL and VLDL. Cholesterol taken up in the form of the individual lipoproteins shows no difference in its availability for the formation of cholesterol ester and surfactant by type II pneumocytes in vitro. Based on the kinetic studies, it appears that VLDL is the major gateway through which cholesterol is provided to satisfy the cholesterol requirements of type II pneumocytes for the synthesis of surfactant. PMID- 9352226 TI - Biological functions of haptoglobin--new pieces to an old puzzle. AB - Haptoglobin, an "acute phase" protein, has different functions, which display genetic polymorphism. The complex of haptoglobin with haemoglobin is metabolized in the heptic reticuloendothelial system. Biosynthesis of haptoglobin occurs not only in the liver, but also in adipose tissue and in lung; providing antioxidant and antimicrobial activity. Changes in the measured concentrations of haptoglobin in serum may help to assess the disease status of patients with inflammations, infections, malignancy etc. (increases) as well as in haemolytic conditions (decreases). Haptoglobin plays a role in stimulation of angiogenesis and has highly potent cholesterolcrystallization-promoting activity. Probably the most important biological function of haptoglobin consists in the host defence responses to infection and inflammation, acting as a natural antagonist for receptor-ligand activation of the immune system. PMID- 9352227 TI - Substance P induces the secretion of gelatinase A from human synovial fibroblasts. AB - We investigated the secretion of the matrix metalloproteinases, interstitial collagenase (matrix metalloproteinase-1), gelatinase A (matrix metalloproteinase 2) and stromelysin-1 (matrix metalloproteinase-3) in human synovial fibroblasts after stimulation with the neuropeptide substance P. Human synovial fibroblasts were stimulated with substance P or interleukin-1 beta (IL-1 beta). In the cell culture media gelatinase A, interstitial collagenase and stromelysin-1 were identified and their activities towards different substrates were determined. Substance P in synovial fibroblasts induced an increase in the overall matrix metalloproteinase activity towards the dinitrophenyl-labelled peptide by 85%, against an increase of 124% after stimulation with IL-1 beta. In case of substance P stimulation, the increase in activity reflects a significantly enhanced secretion of gelatinase A, whereas no significant increase of stromelysin-1 and collagenase secretion could be observed. The matrix metalloproteinase pattern showing the highest gelatinase A secretion was obtained after stimulation with substance P. This pattern was very pronounced and differed very clearly from the pattern seen after IL-1 beta stimulation which caused a significant rise in collagenase and stromelysin-1 activity. We assume that distinct stimulation pathways are involved and that the neuropeptide (substance P), which is always present in the inflamed joint, plays its own and separate role in proliferative processes leading to the cartilage destruction. PMID- 9352228 TI - Effect of cyclosporine A on the release of tissue factor pathway inhibitor from endothelial cells in heart transplant patients and cell culture. AB - We investigated the influence of cyclosporine A on the concentration of tissue factor pathway inhibitor and von Willebrand factor antigen in plasma of heart transplant outpatients. Tissue factor pathway inhibitor was quantified in plasma of blood donors (n = 50) and heart transplant outpatients (n = 50) by a chromogenic substrate assay with a mean of 32.4 micrograms/l and 98.2 micrograms/l, respectively. Von Willebrand factor antigen was determined with an enzyme-linked immunoassay with a mean of 90.9% for blood donors and 184.5% in plasma of heart transplant recipients. In addition, we investigated the effect of cyclosporine A on endothelial cell cultures over an incubation period of four days. A dose-dependent effect of cyclosporine A on the release of endothelial tissue factor pathway inhibitor and von Willebrand factor antigen was determined in a concentration range from 100 to 200 micrograms/l cyclosporine A. The tissue factor pathway inhibitor and von Willebrand factor antigen concentrations in the cell culture supernatant increased during the incubation time according to the cyclosporine A concentration 2-3 fold and 2 fold, respectively. For a further elucidation of the cyclosporine A effect we investigated the influence of cremophor EL, the vehicle of cyclosporine A. Cremophor EL alone did not increase the tissue factor pathway inhibitor release. However, the release was enhanced 2 4 fold after co-stimulation with the calcium ionophore A 23187 (10(-4) mol/l) in a concentration-dependent mode. We conclude that a generalized endothelial damage or activation is most probably caused by cyclosporine A and its vehicle cremophor EL. This process probably depends upon the increase of cytosolic free calcium, as described for the liberation of von Willebrand factor by endothelial cells. PMID- 9352230 TI - The determination of inorganic sulphate in serum and synovial fluid by high performance ion chromatography. AB - A method for the determination of inorganic sulphate based on high performance ion chromatography is presented. The separation was performed on an anion exchange column with a 1.8 mmol/l sodium carbonate/ 1.7 mmol/l sodium hydrogen carbonate-buffer, pH 10.35. Conductivity of the eluate was monitored after suppression of the background conductivity caused by the eluent-buffer. Serum and synovial fluid samples were prepared by ultrafiltration through membranes with a molecular mass cutoff of M(r) 10,000. The viscosity of the synovial fluids was reduced by treatment with hyaluronate lyase before ultrafiltration. The method showed a linear response for sulphate concentrations between 0.5 and 1000 mumol/l. The limit of detection was 1 mumol/l for aqueous standards. For serum the coefficient of variation within-run was 2.3%-2.4%, the coefficient of variation between days 2.9%-3.1%. For synovial fluids the coefficient of variation within-run was 3.1%-3.4%, the coefficient of variation between days 4.6%-5.7%. Standard recovery experiments performed by spiking pools of human sera containing low sulphate concentrations with sulphate concentrations between 5 mumol/l and 40 mumol/l showed recoveries between 98.9% and 100.6%. The corresponding experiments with pools of synovial fluids showed recoveries of 98.3% to 100.9%. As determined from 127 serum samples the reference range for sulphate was 262 mumol/l-420 mumol/l, with a mean value of 314 mumol/l. No dependence on age or sex was observed. The sulphate concentration in 36 synovial fluids from knees affected by inflammatory processes showed a mean value of 424 mumol/l and a standard deviation of 70 mumol/l. In 41 synovial fluids from knees affected by chronic degeneration joint disease, the sulphate concentrations were statistically significantly lower, with a mean of 374 mumol/l and a standard deviation of 58 mumol/l. The concentrations of sulphate in the synovial fluids were statistically significantly higher than those in the serum samples used for determination of the reference range. Following the oral application of a subtoxic single dose of acetaminophen (32.5 mg/kg body weight-62.5 mg/kg body weight) to 4 healthy volunteers, there was a significant decrease in the concentration of sulphate in serum with a minimum at 4-5 h after application of the drug. The cumulative concentration decrease of sulphate in serum and the kinetic constant of the sulphate depletion were not correlated with the applied acetaminophen dose normalized for body weight. PMID- 9352229 TI - Determination of advanced glycation end products in serum by fluorescence spectroscopy and competitive ELISA. AB - Recent studies suggest that advanced glycation endproducts play an important role in cardiovascular complications of ageing, diabetes and end-stage renal failure. Since highly elevated levels of advanced glycation endproducts are present in serum of patients on maintenance haemodialysis, an accurate and rapid assay for their determination would be useful. This would be particularly valuable for monitoring the removal of advanced glycation endproducts by novel dialysis membranes, as well as the effect of new drugs for the inhibition of their formation. Measurement of advanced glycation endproducts in serum was performed by two competitive ELISAs, using a monoclonal antibody directed against imidazolone, an advanced glycation endproduct formed by the reaction of arginine with 3-deoxyglucosone, and a polyclonal antibody directed against keyhole limpet haemocyanin-advanced glycation endproduct, as well as by quantitative fluorescence spectroscopy. Each of the assays showed significant differences between the controls and the maintenance haemodialysis patients. Advanced glycation endproduct levels determined by each of the ELISAs correlated with total and protein-bound fluorescence, but not with each other, suggesting a variable distribution of advanced glycation endproducts on serum proteins among the maintenance haemodialysis patients. PMID- 9352231 TI - Determination of total homocysteine in human plasma by isocratic high-performance liquid chromatography. AB - A simple, sensitive and precise isocratic HPLC method for the determination of total homocysteine in human plasma is described. The thiol compounds were liberated from plasma proteins by reduction with tri-n-butylphosphine and derivatized with a thiol-specific fluorogenic marker, 7-fluoro-benzo-2-oxa-1,3 diazole-4-sulphonate. The derivatives were separated isocratically within 7 min by reversed-phase HPLC using a Superspher 100 RP-18 column as stationary phase. By using this approach more than 200 samples a day can be assayed for total homocysteine. The method was linear up to 100 mumol/l and proved to be sensitive with a detection limit of 0.1 mumol/l and the lowest limit of reliable quantification of 0.5 mumol/l for homocysteine in buffer. Intra- and inter-assay coefficients of variation were both < 4% at a concentration of 10 mumol/l homocysteine. Similar results were obtained for homocysteine concentrations between 0.5 and 100 mumol/l. The analytical recovery for these concentrations ranged from 94.9 to 117.0%. As compared to other protocols published so far, this modified method is less complicated but equally sensitive and reproducible and allows a rapid determination of total homocysteine and cysteine in human plasma under routine conditions. PMID- 9352232 TI - Comparison of urine dipsticks with quantitative methods for microalbuminuria. AB - We describe a new dip- and read dipstick that detects urine albumin at concentrations of 10 mg/l and above and urine creatinine at concentrations of 300 mg/l and above. The albumin assay is based on a high-affinity, dye-binding technique while the creatinine assay is based on the peroxidase-like activity of copper creatinine complexes. With these two-test dipsticks, urines from normal adults supplemented with albumin and creatinine were correctly identified to within +/- 15% of the expected value for both analytes; the between-day coefficients of variation ranged from 7.1% to 16.1%. We tested 275 patients' unmodified urines by the Bayer and Boehringer Mannheim Micral-Test albumin dipsticks and for albumin with the Beckman Array on the same specimens. We also analyzed 42 selected urines from the group of 275 for albumin by another quantitative immunochemical method and by electrophoresis plus a total protein method to estimate the albumin concentration. The quantitative immunochemical methods appear to underestimate the urine albumin concentrations; in these 42 urines measured as negative, i.e., < ca. 16-20 mg/l, by one of the quantitative method but positive by the Bayer dipstick, 33 of these were positive by the electrophoresis/total protein assay combination. The Bayer albumin dipstick correctly identified urines as having < 16 mg/l or > or = 16 mg/l at an 80% rate. At a cutoff of 20 mg/l, the rate increased to 87%. We also determined the urinary albumin/creatinine ratios on the 275 patients using the Bayer two-pad dipstick and found agreement 84% of the time with the same ratio obtained from a quantitative immunochemical method for albumin and a rate-Jaffe method for creatinine; an albumin/creatinine ratio (mg/g) of 30 was used as the discrimination point. Albumin stability studies performed on the Beckman Array patients with six fresh urines showed small but consistent decreases at -20 degrees C but not at 4 degrees C after one month of storage. The albumin in contrived urines, as estimated by electrophoreses/total protein and by the dipsticks did not change at these storage conditions. Boric acid at 1 g/l as a urine preservative had no effect on the measurement of albumin by any of the methods described here nor of the assay of creatinine. Other urinary proteins present at abnormal excretion rates did not interfere with the Bayer albumin dipstick. Abnormal concentrations of bilirubin, citrate, creatine, ascorbic acid, albumin, hemoglobin and myoglobin in urine did not interfere with the creatinine dipstick measurements. The first four of the above did not affect the Bayer dipstick results for albumin. PMID- 9352233 TI - Falsely high ionized magnesium results by an ion-selective electrode method in severe hypomagnesemia. AB - Changes in serum total and ionized magnesium (Mg and Mg2+) and calcium (Ca and Ca2+) were monitored in three patients who transiently developed severe (total Mg < 0.50 mmol/l) to profound hypomagnesemia (total Mg < 0.35 mmol/l) due to cisplatin or interleukin-2 therapies. Mg2+ and Ca2+ were measured with the Nova ion-selective electrodes at 37 degrees C and all results were normalized to pH 7.40. Independent of the etiology, the Mg2+ fraction (Mg2+/total Mg) increased as the concentration of the serum total Mg decreased in all three patients. When the total Mg was around or below 0.35 mmol/l the Mg2+ approached or exceeded total Mg, suggesting an error in the measurement of Mg2+. The findings were extended by including a group of 31 additional patients whose serum total Mg, Mg2+, total Ca, and Ca2+ concentrations varied from abnormally low to above normal. The serum total and ionized concentrations strongly correlated for both Mg (r2 = 0.88) and Ca (r2 = 0.92). The Mg2+ fraction rapidly increased with a fall in the total Mg concentration (r2 = 0.76) and total Mg/total Ca ratio (r2 = 0.71). In fact, with decreasing total Mg concentrations or total Mg/total Ca ratios, the Mg2+ fraction progressively increased to 93-128% of the total, confirming an error in the Mg2+ determinations. The Ca2+ fraction showed a slight and insignificant decrease with falling total Ca concentrations and total Mg/total Ca ratios. The Mg2+ concentration was directly related (r2 = 0.62), whereas the Ca2+ concentration showed a complex relationship to the total Mg/total Ca ratio. Whether this latter relationship represents a technical artifact or a true biological phenomenon requires further study. The apparent overestimation of Mg2+ at very low total Mg concentrations, and in the presence of a very low total Mg/total Ca ratio, could be due to improper chemometric correction of the Ca effect on the Mg electrode, non-linearity, and inadequate calibration. Whatever the mechanism, the failure of this method to correctly measure very low serum Mg2+ concentrations in the sera of patients with severe hypomagnesemia, or likely in any patient with an unusually low total Mg/total Ca ratio, erodes its diagnostic usefulness. PMID- 9352234 TI - Evaluation of the VALAB expert system. AB - The validation of a clinical laboratory report is a process that guarantees the results contained in the report have been obtained under satisfactory metrological conditions and that they are compatible with the information available on the patient. This validation is generally carried out manually by a clinical laboratory professional, but also may be done by an expert system properly programmed, such as the VALAB system. The evaluation presented in this article consists of comparing human and system decisions of validation for 500 randomly selected clinical laboratory reports from hospitalized patients. In this evaluation, 84.8% of the reports examined by the VALAB are accepted directly without any human aid, and only 15.2% require examination by clinical biochemists. PMID- 9352235 TI - Multicentric reference values: shared reference limits. AB - In order to obtain shared reference limits, three laboratories in the same geographical area with a homogeneous population have developed a proposal to produce multicentric reference values. The strategy simulates a virtual laboratory, actually formed by the laboratories involved; the reference limits produced in the virtual laboratory are in fact derived from the blend of reference values obtained by each laboratory. Each laboratory has chosen its own reference sample and has measured the biochemical quantities under study. Reference individuals (n = 171) and 15 biochemical quantities among the most measured in clinical laboratories were selected. The reference values obtained in each laboratory were blended when permitted by the Harris & Boyd test (Clin Chem 1990; 36:265-70). The multicentric reference limits obtained by the virtual laboratory for each quantity were estimated according to the recommendations of the International Federation of Clinical Chemistry. For each quantity, each laboratory, with the results observed in their reference sample, estimated the diagnostic specificity, using as cut-off values the corresponding multicentric reference limits. Each observed value of diagnostic specificity was compared with the theoretical diagnostic specificity value, equal to 0.975, that should be observed when a reference limit is used as cut-off value. The multicentric reference limits obtained by the virtual laboratory are valid in all cases with the exception of the upper reference limit for the concentrations of calcium(II) and urate in serum in one of the laboratories. PMID- 9352236 TI - Current stage of standardization of measurements of specific polypeptides and proteins discussed in light of steps needed towards a comprehensive measurement system. AB - We present a standardization model for the measurement of specific polypeptides and proteins, which is based on an integrated development of all important elements of a reference measurement system. Generally, the model is in line with other current recommendations. However, it puts special emphasis on the definition of the analyte and on the role of reference methods for verification of the standardization process by measurement of patient specimens. Further, we discuss the needs for its implementation in the routine laboratory. In the light of this model, we investigate the current stage of standardization of routine methods for enzymes, peptide hormones, proteins, apolipoproteins, glycohaemoglobin, and tumour markers. PMID- 9352237 TI - Harris & Boyd's test for partitioning the reference values. PMID- 9352238 TI - Storage of serum for the determination of ionized magnesium. PMID- 9352239 TI - Viruses and cancers: possible role of hepatitis C virus. AB - Oncogenesis is a multifactorial process in which environmental, genetic and infectious factors are variably involved. A possible role of specific viruses has been suggested in at least 15% of human cancers. Hepatitis C virus (HCV), which is both hepato- and lymphotropic, is responsible for various liver disorders, i.e. chronic hepatitis, cirrhosis and hepatocelluar carcinoma, as well as for a constellation of extrahepatic immune-mediated manifestations, among which is mixed cryoglobulinaemia. This is a systemic disorder secondary to a chronic, benign B-lymphocyte proliferation, which in some subjects may evolve to a malignant non-Hodgkin's lymphoma (NHL). Interestingly, recent studies reported the appearance of malignant B-cell neoplasias in patients with type C chronic hepatitis; moreover, in a significant number (from 22% to 50%) of 'idiopathic' NHLs, the presence of HCV infection has been demonstrated. The presence of a geographical etherogeneity in the prevalence of HCV-positive NHL suggests that other co-factors, i.e. genetic and environmental, could be involved in the lymphomagenesis. HCV may exert its oncogenic potential in two different directions, leading to liver cancer or B-cell lymphoma. PMID- 9352240 TI - Multiple biochemical effects in the pathogenesis of alcoholic fatty liver. AB - The pathogenesis of alcoholic fatty liver is unknown, but several causes have been proposed based on biochemical findings. These include the metabolism of alcohol leading to a shift in the cytosolic [NAD+]/ [NADH] ratio to reduction, which in turn causes a direct inhibition of beta-oxidation and enhanced triacylglycerol formation via the [glycerol-3-phosphate]/[dihydroxyacetone phosphate] ratio. There are also chronic effects of ethanol on hepatic enzyme activities. Thus, increased activity of phosphatidate phosphohydrolase, an increased amount of fatty acid binding protein, decreased secretion of very low density lipoprotein and impairment of the respiratory chain as a result of decreased protein synthesis or decreased amounts of ubiquinone could all lead to fat accumulation and steatosis. The interplay of each of these with nutritional and genetic factors would then lead to the heterogeneity of the severity and characteristics of the steatosis observed in human alcoholics. PMID- 9352241 TI - Elevated plasma levels of reduced homocysteine in common variable immunodeficiency--a marker of enhanced oxidative stress. AB - Based on previous studies from our group, we hypothesized that enhanced oxidative stress in association with a persistent immune activation may be important in both the immunopathogenesis and certain clinical manifestations in a subgroup of patients with common variable immunodeficiency (CVI). To explore this hypothesis further, we examined plasma levels of lipid peroxidation, antioxidant vitamins and redox status of various thiol species in 20 CVI patients and 16 healthy control subjects. We found significantly higher malondialdehyde (MDA) levels in plasma from CVI patients than in healthy control subjects. Furthermore, in a subgroup of CVI patients characterized by persistent immune activation in vivo (CVIHyper), we found significantly decreased levels of vitamin E and beta carotene. In the CVI patients, there was a significant inverse correlation between MDA levels and levels of vitamin E and beta-carotene. Finally, we found a marked elevation in plasma levels of reduced homocysteine in the CVI group, but no corresponding rise in plasma levels of total homocysteine. In the CVI group, the high plasma levels of reduced homocysteine were significantly correlated with enhanced lipid peroxidation and low levels of vitamin E. The results of the present study further support a role for enhanced oxidative stress in the immunopathogenesis of CVI. Furthermore, our finding of markedly elevated plasma levels of reduced homocysteine in CVI patients without simultaneous elevation of other homocysteine species suggests that this disturbance in homocysteine metabolism may be related to enhanced oxidative stress. PMID- 9352242 TI - Oxidative stress in immunodeficiency. PMID- 9352243 TI - Stress-induced blood pressure measurements predict left ventricular mass over three years among borderline hypertensive men. PMID- 9352244 TI - Lipoprotein lipase gene polymorphisms in ischaemic stroke and carotid stenosis. AB - Ischaemic stroke is pathogenetically heterogeneous, but there is strong evidence that genetic as well as environment factors contribute to the risk of the individual. Here we report the similar distribution of polymorphic markers of the lipoprotein lipase (LPL) gene in 128 patients with ischaemic stroke, 56 patients with carotid artery stenosis and 95 healthy control subjects, in spite of a significant influence of the Asn291-->Ser mutation on serum levels of triglycerides. We conclude that these LPL polymorphisms do not contribute greatly to the overall risk of ischaemic stroke in the general population. PMID- 9352246 TI - Ocular and systemic reactivity to isoprenaline in patients with insulin-dependent diabetes mellitus. AB - There is experimental evidence of decreased beta-adrenergic myocardial sensitivity in patients with insulin-dependent diabetes mellitus (IDDM). In the present study we hypothesized that the ocular response to isoprenaline, as a consequence of increased arterial vessel rigidity, might also be blunted in patients with IDDM. We therefore compared the correlation between systemic pulse pressure amplitude (PPA) and fundus pulsation amplitude (FPA) during intravenous isoprenaline administration in 11 otherwise healthy IDDM patients and 11 healthy control subjects. Ocular fundus pulsations were measured by a recently developed laser interferometric method. Isoprenaline increased PPA in both study groups in a dose-dependent way, but the response was significantly less in IDDM patients (at 0.8 microgram min-1: +38% in control subjects, +27% in IDDM patients, P < 0.05 between groups). Moreover, a dose-dependent increase in FPA was observed, which again was more pronounced in healthy subjects (at 0.8 microgram min-1: +45% in controls, +17% in IDDM patients, P < 0.005 between groups). The regression line between PPA and FPA was very close to the 45 degrees line in healthy subjects, whereas it was significantly flattened in IDDM patients. In conclusion, linear regression between PPA and FPA during isoprenaline suggests arterial stiffening in patients with IDDM. Hence, comparison of systemic PPA and FPA during isoprenaline provocation may be a useful method of estimating changes in arterial capacitance in patients with diabetes mellitus. PMID- 9352245 TI - Human uterine smooth muscle exhibits a very low phosphocreatine/ATP ratio as assessed by in vitro and in vivo measurements. AB - The purpose of the study was to investigate by in vitro and in vivo methods the phosphocreatine (PCr)/ ATP ratio as an expression of the energy metabolic state of human myometrium in comparison with striated skeletal muscle. The contents of PCr and adenylates in biopsies of uterine smooth muscle and m. rectus abdominis from seven term pregnant women were determined in vitro and compared with results obtained in vivo by phosphorus magnetic resonance spectroscopy (31P-MRS) in the uterus and m. gastrocnemius of eight non-pregnant women. The PCr/ATP ratio in the striated skeletal muscle was about three times higher than that of the myometrium. The results of the in vitro biopsy part of the study and the in vivo 31P-MRS part conformed with each other. In the biopsies both PCr and ATP concentrations were significantly lower in the myometrium than in the rectus muscle, but the difference for PCr was more pronounced, accounting for the significantly lower PCr/ATP ratio in the uterine smooth muscle. The energy metabolic pattern of uterine smooth muscle differs from that of striated skeletal muscle regarding the contents of high-energy phosphocompounds and the PCr/ATP ratio. This in vivo finding is the first report on human smooth muscle using 31P MRS. PMID- 9352247 TI - Microalbuminuria and renal haemodynamics in essential hypertension. AB - The present study was designed to evaluate the renal haemodynamic pattern of never-treated microalbuminuric and normoalbuminuric patients with essential hypertension. A total of 19 never-treated essential hypertensive patients with microalbuminuria were selected and, as control subjects, 24 never-treated essential hypertensive patients without microalbuminuria (determined on three 24 h urine collections) were recruited. In the two groups, we compared blood pressure values, standing plasma noradrenaline, plasma renin activity, plasma aldosterone, urinary aldosterone, lipid profile, serum glucose and uric acid, glomerular filtration rate and renal plasma flow. In comparison with normoalbuminuric patients, microalbuminuric patients showed significantly higher systolic blood pressure values (P < 0.05), higher renal vascular resistances (P < 0.05) and lower plasma renin activity values (P < 0.01). Urinary albumin excretion showed a significant positive correlation with systolic (r = 0.46, P < 0.005) and mean blood pressure (r = 0.38, P < 0.05), serum uric acid (r = 0.43, P < 0.005) and triglyceride values (r = 0.36, P < 0.005), and a significant negative correlation with plasma renin activity (r = -0.34, P < 0.05). The present data are consistent with the occurrence of renal vasoconstriction in microalbuminuric never-treated essential hypertensive patients. PMID- 9352248 TI - Both plasma and renal endothelin-1 participate in the acute cardiovascular response to exercise. AB - Plasma endothelin (ET-1) and renal endothelin are two distinct functional systems involved in maintaining blood volume. To investigate whether plasma and renal ET 1 participate in the cardiovascular response to exercise-induced hypovolaemia, we studied changes in plasma and urinary ET-1 in healthy non-professional athletes after 2 h of jogging performed both without and with drinking isotonic fluids. After the run, which caused a 13% plasma volume (PV) reduction, plasma and renal ET-1 (+117% and +118%) increased significantly (all P < 0.01). Fluid loss restitution during the run significantly attenuated either the PV contraction ( 1.2%) and plasma and renal ET-1 increase (+2 and +3%). At multiple regression analysis changes in AVP plasma concentration, and not in PRA or PV per se, were significantly related to ET-1 changes both in plasma and urine. The present findings indicate that both plasma and renal ET-1 participate in the cardiovascular response to hypovolaemia induced by long-lasting, dynamic exercise. PMID- 9352249 TI - Glycaemic control and in vivo non-oxidative Maillard reaction: urinary excretion of pyrraline in diabetes patients. AB - The presence of pyrraline in human urine has recently been described. Using reversed-phase high-performance liquid chromatography, we measured urinary pyrraline in 45 insulin-treated diabetic patients with preserved renal function and in 30 age- and sex-matched healthy subjects. The relationship between urinary pyrraline and metabolic control parameters in the diabetic population (glycaemia, fructosamine, haemoglobin A1c, and 1-year mean haemoglobin A1c) was evaluated. The mean urinary level of pyrraline in diabetic patients with poor glycaemic control (HbA1c > 9.5%) was higher than that in healthy subjects (1.12 +/- 0.35 vs. 0.75 +/- 0.2 mumol mmol-1 creatinine, P < 0.04), whereas in patients with good to moderate glycaemic control (HbA1c < 9.5) it was slightly but not significantly higher than in healthy subjects (0.80 +/- 0.3 mumol mmol-1 creatinine vs. 0.75 +/- 0.2 mumol mmol-1 creatinine). There is a significant correlation between urinary pyrraline level and glycaemia (P < 0.008), haemoglobin A1c (P < 0.01) and 1-year mean haemoglobin A1c values (P < 0.007), but not with fructosamine. The results of the present work prove, for the first time, that glycaemic status influences circulating levels of advanced Maillard reaction products. PMID- 9352250 TI - Influence of family history of hypertension on insulin sensitivity in lean and obese hypertensive subjects. AB - We evaluated the influence of family history of hypertension on insulin sensitivity in lean and obese hypertensive subjects (H): 40 lean [body mass index (BMI) < or = 25 kg m-2] H with normotensive parents (F-), 50 lean H with one or two parents hypertensive (F+), 30 obese HF- (BMI > or = 30 kg m-2) and 35 obese HF+. The four groups were comparable in terms of age, sex and ambulatory blood pressure values. We evaluated glucose, insulin and C-peptide before and 30, 60, 90 and 120 min after an oral glucose load, insulin sensitivity index (ISI, fasting glucose/insulin ratio), fasting insulin/C-peptide ratio (I/Cp). Glucose, fasting and during test, and I/Cp were similar among the four groups; insulin and C-peptide, fasting and stimulated, were significantly higher and ISI lower in obese H than in lean H; at similar BMI, insulin and C-peptide were significantly higher in F+ than in F-. Insulin directly correlated with night-time blood pressure only in lean HF-. The correlation between insulin and BMI was significantly closer in F-than in F+. In conclusion, family history of hypertension appears to play a relevant role in insulin sensitivity in hypertensive subjects also in the presence of obesity. PMID- 9352251 TI - Effects of dietary fat saturation on eicosanoid production, platelet aggregation and blood pressure. AB - The effects of dietary fat saturation on eicosanoid urinary excretion, platelet aggregation (PA) and blood pressure (BP) were studied in 42 healthy subjects. They consumed four consecutive diets differing in their fat saturation [saturated (SFA); monounsaturated (MUFA); polyunsaturated n-6 (PUFA n-6); and polyunsaturated n-6/n-3, (PUFA n-3)]. Each diet period lasted 5 weeks. There were no differences in 24-h 2,3-dinor-6- keto-prostaglandin F1 alpha excretion among dietary periods. A significant effect was noted regarding the excretion of 11 dehydro-thromboxane B2 (P < 0.0001). During the PUFA n-6 phase the excretion was significantly higher than during SFA and MUFA periods. Dietary fatty acid composition had a significant effect on ADP (1 mumolL-1) and collagen (2 mgL-1) induced PA. Dietary fat also had a significant effect on systolic and diastolic blood pressure (P < 0.0001). Both were significantly higher during the SFA period than during the other three periods. Our findings suggest that changes in dietary fatty acids may have mild, but significant, effects on eicosanoid production, platelet aggregation and blood pressure. PMID- 9352252 TI - Effects of the angiotensin-converting enzyme inhibitor enalapril on blood haematopoietic progenitors and acetyl-N-Ser-Asp-Lys-Pro concentrations. AB - Acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) is a physiological inhibitor of the proliferation of haematopoietic stem cells. In 12 healthy volunteers treated with the angiotensin-converting enzyme (ACE) inhibitor enalapril (20 mg day-1 for 15 days), we studied plasma and urinary AcSDKP levels, the in vitro degradation of AcSDKP by plasma ACE and the numbers of circulating haematopoietic progenitors (granulocyte-monocytic colony forming unit: CFU-GM; burst forming unit-erythroid: BFU-E; and mixed colony forming unit: CFU-mixed). During treatment, plasma and urinary AcSDKP concentrations increased 2- to 5-fold, degradation of AcSDKP was reduced, and CFU-mixed significantly increased by 100% while BFU-E and CFU-GM significantly decreased by 16% and 26%, respectively. These results indicate that ACE inhibitors may be of value during chemotherapy or radiotherapy, warranting further study. PMID- 9352253 TI - Muramic acid in human peripheral blood leucocytes in different age groups. AB - Presence of muramic acid (as a marker for bacterial cell wall peptidoglycan) was analysed by gas chromatography and mass spectrometry in the peripheral blood leucocytes of subjects from a range of ages (9-80 years) and groups (healthy individuals, patients with rheumatoid arthritis, osteoarthrosis, essential hypertension or multiple sclerosis). Sixty per cent of the sample from the youngest subjects contained detectable muramic acid. The percentage of people with circulating leucocytes containing muramic acid decreased gradually with age, being less than 5% in all groups over 40 years. No clear correlation between the presence of muramic acid and the disease was observed. PMID- 9352254 TI - Diphtheria immunity in Flanders. AB - A serological survey to determine the immunity to diphtheria in the Flemish population was conducted according to the recommendations of the World Health Organization. Immunity to diphtheria was determined on a randomised, stratified sample (1679 serum samples) from an existing serum bank (4058 serum samples) representative of the Flemish population. All age groups between 0 and 100 years were included. A tissue (Vero cell) culture toxin neutralisation assay was used to measure serum diph-theria antitoxin concentrations. The results showed that 43% of the Flemish population was protected against diphtheria (antitoxin titre, > or = 0.1 IU/ml), while 32% was susceptible (antitoxin titre, < 0.01 IU/ml); for 25%, protection was of limited duration (antitoxin titre, > or = 0.01 IU/ml and < 0.1 IU/ml). The proportion of susceptible subjects showed a significant age related increase, with the highest values in the 35 to 44 and 45 to 54 age groups (57.9% and 55.5%, respectively). These results emphasise the need for booster immunization of adults. PMID- 9352255 TI - Diagnostic laparoscopy in patients with acute leukemia and suspected hepatic candidiasis. AB - To assess the value of laparoscopy in the diagnosis of suspected hepatosplenic candidiasis in patients with acute leukemia, a retrospective analysis of 28 laparoscopies was conducted. In all but two cases, imaging of the liver showed focal lesions before laparoscopy. Diagnosis of hepatic candidiasis was established significantly more often when the biopsy was targeted at white nodules (in 12 of 22 laparoscopies) than when targeted randomly or at scars (0 of 6 laparoscopies) (p = 0.017, chi-square test). Yeast was detected more often if the laparoscopy was performed during the three-week period after recovery from neutropenia (in 8 of 12 laparoscopies) than when performed later (in 4 of 16 laparoscopies) (p = 0.028, chi-square test). In addition to the 12 laparoscopically diagnosed patients, eight (29%) patients were diagnosed with disseminated Candida infection by other methods. In another eight (29%) patients the causative agent was not identified. No bleeding or other problems occurred after the laparoscopy. Laparoscopy-guided liver biopsy is most useful if biopsies are targeted to macroscopic lesions and if laparoscopy is performed soon after recovery from neutropenia. PMID- 9352256 TI - Pneumococcal resistance patterns in Europe. AB - The emergence of Streptococcus pneumoniae strains with decreased susceptibility to penicillin has been reported worldwide over the past 20 years. However, there are striking discrepancies in penicillin susceptibility among various European countries, suggesting that local conditions may affect clonal propagation or de novo selection of resistant strains. In the present study, data on penicillin resistance patterns, antibiotic use and mode of administration, and treatment compliance in five European countries (France, Spain, Germany, Italy, and the UK) were compared. High prevalence rates of penicillin-resistant pneumococci have been reported in Spain and France, where antibiotics are widely prescribed, and overall in Europe, patient compliance with more than 50% of oral antimicrobial prescriptions is inadequate. The low prevalence of penicillin resistance in Germany and the UK coincides with lower antibiotic consumption and better treatment compliance in these countries. Recent attempts to raise public awareness and to restrict and improve indications for antimicrobial agents have resulted in decreased pneumococcal resistance in Hungary and Iceland, suggesting that pneumococcal resistance can be reversed. PMID- 9352257 TI - Evaluation of commercial slides for detection of immunoglobulin G against Bartonella henselae by indirect immunofluorescence. AB - Four commercial slides were compared with in-house slides for the detection of immunoglobulin G (IgG) against Bartonella henselae in 58 healthy persons from a rural region by an indirect immunofluorescence assay. MRL-BA slides (MRL Diagnostics, USA) and Virion slides (Virion, Switzerland) with agar-derived Bartonella henselae showed IgG titers of > or = 1:256 in 44.8% and 51.7%, respectively, whereas Bion slides (Bios, Germany), MRL-Vero slides (MRL Diagnostics), and in-house slides with cell-associated Bartonella henselae showed such titers in 3.4%, 5.1% and 3.4%, respectively. The MRL-Vero slides (Bartonella IgG substrate slides, MRL Diagnostics) were further evaluated with 26 patients with cat scratch disease, 20 patients with lymphadenopathy not due to cat scratch disease, 100 blood donors from an urban area, and 120 blood donors from a mixed urban/rural area. In our mixed urban/rural population the IgG titer of 1:256 had a sensitivity of 84.6% and a specificity of 93.4% for the serodiagnosis of cat scratch disease. Seroprevalence was higher in blood donors from the mixed area (50.8%) than from the urban area (37%). MRL-Vero slides were considered useful for the serodiagnosis of cat scratch disease by indirect immunofluorescence and have replaced our in-house system. However, patients with low IgG titers should be retested three to four weeks after initial sampling to demonstrate a possible rise of IgG titers in paired sera. PMID- 9352259 TI - Correlation between antiretroviral resistance mutations, biological parameters, and clinical evolution in zidovudine-treated patients infected with human immunodeficiency virus type 1. AB - To evaluate the correlation between zidovudine (ZDV) resistance mutations of human immunodeficiency virus type 1 (HIV-1), biological parameters, and clinical evolution, 111 HIV-1-infected patients treated with ZDV were studied. Specific mutations at codons 70, 215, and 41 in the HIV-1 reverse transcriptase coding region conferring resistance to ZDV were detected using a selective polymerase chain reaction. The appearance of ZDV resistance mutations was significantly correlated with baseline clinical stage, CD4+ cell count, and viral load, but not with duration of ZDV therapy or p24 antigen level. In univariate analysis, results showed a prognostic role of mutations at codons 215 and 41 for clinical progression to the acquired immune deficiency syndrome (AIDS) or death. In multivariate analysis after controlling for viral load, CD4+ cell count, and clinical stage, the presence of the mutation at codon 215 (but not at codon 41) remained an independent predictor of subsequent clinical evolution. PMID- 9352258 TI - Evaluation of a novel immunoglobulin A capture enzyme immunoassay for diagnosis of cytomegalovirus infection in renal and heart transplant recipients. AB - In a retrospective cohort study of 68 organ transplant patients, the usefulness of a new commercial immunoglobulin A (IgA) antibody capture enzyme immunoassay (EIA) specific to human cytomegalovirus (CMV) for the early diagnosis of CMV disease was investigated. The results were compared with those obtained with the CMV pp65 antigen assay in peripheral blood leukocytes, an IgM immunoblot assay, and six other commercial EIAs. In 21 of 28 patients with CMV disease, the pp65 antigen assay and the immunoblot assay identified patients before the onset of disease more frequently than any other serological test method (17 and 13 patients, respectively; p = 0.0029). In patients at risk for primary CMV infection, the pp65 antigen assay was the only method that identified all patients prior to the onset of CMV disease (p = 0.008). For the other patients who were at risk for CMV infection, the pp65 antigen assay and the immunoblot assay detected infection before CMV disease more frequently than any other test system (p = 0.026). With respect to CMV disease, both immunoblotting and the pp65 antigen assay showed excellent sensitivity (100% and 89%, respectively). However, the specificity of the immunoblot was poor (41%), while the specificity of the pp65 antigen assay was reasonably good (68%). The IgA capture EIA showed moderate sensitivity (61%) and reasonable specificity (76%). In conclusion, the pp65 antigen assay, which detects pp65 antigen in leukocytes, is the method of choice for diagnosis of either primary or recurrent CMV infection. The specificity of the pp65 antigen assay was improved by additional testing for specific IgA and IgM antibodies (95% vs. 68%). The IgA assay is of limited value in renal and heart transplant patients, since it detected IgA antibodies only sporadically, and even then, too late for a timely therapy. PMID- 9352260 TI - Multicenter clinical comparison of resin-containing bottles with standard aerobic and anaerobic bottles for culture of microorganisms from blood. AB - In a study comparing the Bactec 9240 (Plus Aerobic/F and Anaerobic/F bottles, containing resins; Becton Dickinson, USA) and the Vital (standard aerobic and anaerobic bottles, with no additives; bioMerieux, France) blood culture systems, 6456 sets of four bottles of 9660 blood cultures submitted were evaluated. There were 531 clinically significant isolates from 795 positive blood cultures. Of the 531 positive blood cultures, 355 were positive in both systems, 141 with the Bactec 9240 alone, and 30 with the Vital alone (p < 0.001); five were not detected by either system. The average time to detection of positive cultures for the matched sets was 10.65 h and 18.41 h by the Bactec 9240 system and the Vital system, respectively. The false-positive rate per bottle was 0.65% in the Bactec 9240 and 0.71% in the Vital. The rate of false-negative pairs (i.e., major errors) was very low (0.12% for the Bactec 9240, 0.19% for the Vital) and not significantly different between the two systems. The striking differences in recovery of microorganisms may be due to the presence of resins in the Bactec medium. However, the observed superiority of the Bactec 9240, even for patients not receiving antibiotics, suggests that resins adsorb other inhibitors present in patients' blood. PMID- 9352261 TI - Eosinophilia in patients infected with the human immunodeficiency virus. AB - The prevalence and significance of peripheral blood eosinophilia in patients infected with the human immunodeficiency virus (HIV) were evaluated. Fifteen of 119 consecutive patients had absolute eosinophil counts of > 450/mm3. During a mean follow-up period of 419 days eosinophilia could be identified as secondary to a parasitic infection in only one patient. Correlation with disease stage showed a higher rate of advanced disease in patients with absolute eosinophilia. In a multivariate regression analysis, only low CD4+ cell counts, not the CDC disease stage or the use of antiretroviral therapy or primary prophylaxis, contributed significantly to the prevalence of eosinophilia. It is concluded that expensive laboratory investigations in asymptomatic patients with advanced-stage HIV disease are neither necessary nor cost effective. PMID- 9352262 TI - Acute hepatitis associated with Campylobacter jejuni bacteraemia. AB - A case of acute hepatitis associated with Campylobacter jejuni bacteraemia is reported. Transaminase levels were increased over 50-fold in a patient with clinical features of enteritis and septicaemia. Campylobacter jejuni was isolated from blood and faecal cultures. Other infective and noninfective causes of acute hepatitis were excluded. The patient's symptoms and liver function values improved after antimicrobial therapy. Hepatitis should be considered as a complication of human Campylobacter jejuni infection. PMID- 9352263 TI - Lactobacillus species as emerging pathogens in neutropenic patients. AB - The intensive use of broad-spectrum antibiotics in the context of prolonged and severe neutropenia has contributed to the emergence of unusual pathogens. Four new cases of severe Lactobacillus infections-three of septicemia and one of pneumonia-are reported. They occurred in patients with acute leukemia who had chemotherapy-induced neutropenia. All patients were treated in the same intensive care unit and received the same antimicrobial prophylaxis which included a total bowel decontamination containing vancomycin. The four patients were treated with a combination of intravenous ceftazidime and vancomycin prior to the development of Lactobacillus infection. Improvement in the condition of all patients was obtained with a treatment including penicillin and concurrent recovery of granulopoiesis. PMID- 9352264 TI - Evaluation of a new commercial microimmunofluorescence test for detection of antibodies to Chlamydia pneumoniae, Chlamydia trachomatis, and Chlamydia psittaci. AB - A new commercial test for chlamydial serology, the MRL-Micro-Immunofluorescent Test (MRL; MRL Diagnostics, USA) was compared with the standard microimmunofluorescence test (MIF) using sera from 246 patients. Chlamydia pneumoniae immunoglobulin G (IgG) antibodies were detected in 46.3% (MIF) and 64.2% (MRL) of sera and Chlamydia trachomatis IgG in 23.2% (MIF) and 25.2% (MRL); Chlamydia psittaci IgG antibodies were found with the MRL in 1% of the sera from a general population and in 17.3% of preselected sera with elevated complement fixation titers. Titers were usually higher with the MRL. IgG titers of > or = 1:512 were detected in only 2% of sera using the standard MIF but in 30% using the MRL. In 16 sera from three Chlamydia pneumoniae culture-positive patients, the diagnosis of acute infection could be confirmed serologically in one with the MRL test but in none with the MIF test, indicating a higher sensitivity of the MRL. PMID- 9352265 TI - Simple enzymatic method for rapid identification of a Staphylococcus aureus subspecies aureus biovar. AB - In order to correctly identify a new biovar of Staphylococcus aureus subsp, aureus, (NBSA) a simple, rapid, and reliable enzymatic assay was developed. The assay was based on the detection of the production of three enzymes: alpha glucosidase, beta-glucosidase and beta-N-acetyl-glucosaminidase. Of a total of 46 isolates of Staphylococcus aureus subsp. aureus from clinical specimens, the new assay correctly identified 19 as NBSA and 27 as typical Staphylococcus aureus subsp. aureus. Among the 19 NBSA isolates, 15 (79%) showed a clear biochemical profile while only four isolates (21%) showed a less well-defined enzymatic combination, due to a phenotypic alteration caused by subculturing. Since this assay is both simple to perform and inexpensive, it is potentially applicable in the laboratory. PMID- 9352266 TI - Antifungal susceptibility testing of yeast isolates from blood cultures by microbroth dilution and the E test. AB - The results of microbroth dilution were compared with those of the E test for 169 yeast isolates tested for their susceptibility to antifungal agents. All isolates were tested by both methods against amphotericin B, ketoconazole, fluconazole, and itraconazole. The E test results generally correlated well with those obtained by the reference method. There was at least 80% agreement of minimum inhibitory concentration results within two dilutions for all yeast species and agents tested, except for Cryptococcus neoformans tested with fluconazole (8% agreement). The E test appears to be a suitable alternative antifungal susceptibility test method for yeasts, although improvements are required for testing Cryptococcus neoformans against fluconazole. PMID- 9352267 TI - Candida famata fungemia in a surgical patient successfully treated with fluconazole. PMID- 9352268 TI - Disseminated isosporiasis in an AIDS patient. PMID- 9352269 TI - Influence of lectins on the infectivity of elementary bodies of Chlamydia trachomatis D IC CAL 8 by synovial cells. PMID- 9352270 TI - Transferable resistance to cefotaxime, ceftazidime, and aztreonam and production of extended-spectrum beta-lactamase in a strain of Salmonella enteritidis. PMID- 9352271 TI - Hyperopia correction using an erodible mask excimer laser delivery system coupled to an axicon: preliminary results. AB - PURPOSE: This paper presents the results of the first human trial on the correction of hyperopia using an erodible mask excimer laser delivery system coupled to an axicon. METHODS: We treated 17 eyes of 17 patients (age range 34-62 years) for the correction of +3.21 +/- 1.04 D (range +1.00 to +4.00 D). The hyperopic correction was made using an erodible mask inserted on the laser optical pathway, to produce a circular ablation measuring 6.5 mm in diameter. An axicon was then used to create a blend transition zone from 6.5 mm up to 9.4 mm in diameter. Eyes were evaluated at one, three and six months after surgery. RESULTS: Reepithelization was always observed by the fifth postoperative day, despite the large area of deepithelization (diameter 9.5 mm). Mean refractive error one month after treatment was -2.44 +/- 1.59 D (range 0.00 to -6.50 D). Five eyes (29.4%) had a best corrected visual acuity loss more than two to three lines; all eyes showed mild annular haze not involving the central part of the cornea. Six months after treatment, mean refractive error was -0.88 +/- 0.99 D (range +0.50 to -3.00 D). Compared to preoperative status, 13 eyes (76.5%) showed an improvement in uncorrected distance visual acuity (1-8 lines), and 14 eyes (82.4%) showed an improvement in uncorrected vision at reading distance (3-7 lines). Two eyes (11.7%) showed a best corrected visual acuity loss of two of three lines. CONCLUSIONS: These preliminary results indicate this approach is effective in reducing hyperopia, while its predictability has still to be proved in a larger treatment group with longer follow-up. A cautious approach to this technique is still advisable, especially for higher hyperopic corrections, in view of the large best corrected visual acuity loss seen in two eyes at six months. PMID- 9352272 TI - Eyelid hyperlaxity and obstructive sleep apnea (O.S.A.) syndrome. AB - PURPOSE: An association between the floppy eyelid syndrome and the obstructive sleep apnea syndrome (O.S.A.) has been reported. We studied eyelid tissue elasticity and other ophthalmologic findings in a large number of patients with sleep disorders. MATERIAL AND METHODS: Sixty-nine patients with sleep disorders were evaluated. Two thirds were found to have O.S.A., and one third was treated at night by nasal continuous positive airway pressure (nasal C.P.A.P.). Slit lamp examination, eyelid measurements and Schirmer test were performed. RESULTS: Eyelid hyperlaxity was increased in patients with O.S.A. The floppy eyelid syndrome (associated papillary conjunctivitis), however, was rare. Associated corneal lesions were rare, and most patients were asymptomatic. In some cases, ocular irritation was due to air leaks from nasal C.P.A.P. A significant proportion of patients required treatment for primary open angle glaucoma. CONCLUSIONS: Our study of 69 patients found an association between O.S.A. and eyelid hyperlaxity. PMID- 9352273 TI - Medial canthus tumor surgery: a prospective study of microscopically controlled excision. AB - OBJECTIVES: We set out to demonstrate that medial canthus tumors are malignancies requiring microscopically-controlled excision for a high cure rate. We also aim to show that reconstruction can have good esthetic results with a few simple techniques. METHODS: During 1992, we treated 38 basal cell carcinomas of the medial canthus, employing our own two-step Mohs' surgery. All cases were reconstructed with five simple techniques: "laissez faire", full thickness graft, nasoglabellar flap, mild-line forehead flap or combination of flaps. RESULTS: No recurrent basal cell carcinomas have been observed in our patients during the last four years. All the medial canthus tumours were basal cell carcinomas, eight involving morpheiform infiltration. Perineural infiltration was observed in two cases. CONCLUSIONS: Micrographic surgery for medial canthus malignant tumors is the best resection technique. Infiltrating, basal cell carcinomas, are the most common tumors of medial canthus, but also have an excellent cure rate. Reconstruction with a small number of flaps and skin graft is generally an easy process, producing highly satisfactory results. PMID- 9352274 TI - Causes of enucleation: a clinicopathological study. AB - BACKGROUND: Enucleation is an approach used for unresponsive end-stage ocular disease often resulting in blind, painful or cosmetically unacceptable eyes. METHODS: We reviewed the clinicopathological data on 3506 enucleations performed over a 50-year period, 1945-1995. Histopathological data were divided into eight groups according to the causes leading to enucleation: trauma, phthisis, corneal disease, inflammation, vitreoretinal disease, glaucoma, tumors and infections. RESULTS: The study considered 3506 enucleated eyes of 3482 patients, 2467 (70.8%) males and 1011 (29.1%) females (4 sex unspecified). The z-test showed there were significantly more enucleations in males for phthisis (p < 5.05), infections (p < 0.01), trauma (p < 0.01) and inflammation (p < 0.01) and more enucleations for tumors in females (p < 0.01). There were no differences between males and females with regard to enucleations for glaucoma, vitreoretinal and corneal diseases (p > 0.05). The 0-9 years age group was most frequently affected, accounting for 29.7% of the cases. Patients aged less than 30 years constituted 53.6% of all enucleations. The primary or underlying causes leading to enucleation were tumors (1185 eyes, 33.8%), phthisis (587 eyes, 16.7), glaucoma (561 eyes, 16.0%), vitreoretinal diseases (320 eyes, 9.1%), infections (259 eyes, 7.4%), corneal disease (229 eyes, 6.5%), trauma (209 eyes, 6.0%) and inflammation (156 eyes, 4.4%). Time trends in enucleating eyes with different causes showed the number of enucleations for phthisis, infections, corneal diseases, trauma and inflammations had dropped during the ten-year period 1986-1995 compared to 1976-1985 (z-test, p < 0.01). There were no real changes in enucleations for glaucoma and vitreoretinal diseases and there was an increase in the number of enucleations for tumors (p < 0.01). CONCLUSIONS: Improved diagnostic and therapeutic methods, widespread use of photocoagulation in vascular disorders and vitreoretinal surgery in traumas, effective antimicrobial treatment, increasing use of corticosteroids and immunosuppressants, have contributed to the decreasing frequency of enucleation. Tumor patients generally presented late with advanced tumors totally filling the eye, not salvageable by other non-invasive treatment methods. Prompt diagnosis of intraocular malignant tumors (retinoblastoma and malignant melanoma) may reduce the need for enucleation. PMID- 9352275 TI - Electrophysiological assessment of visual pathways in glaucoma. AB - PURPOSE: To assess nerve conduction in visual pathways in patients with open angle glaucoma. METHODS: Pattern-electroretinograms (PERG) and visual-evoked potentials (VEP) were simultaneously recorded in 16 patients with open-angle glaucoma (POAG) and 15 age-matched controls. The visual stimuli were checker board patterns (the check edges subtend 15'; the contrast was 70% and reversed at the rate of 2 reversals/s). RESULTS: POAG patients showed significantly higher PERG and VEP latencies (ANOVA: P < 0.01) and significantly lower amplitudes than controls; the retinocortical time (RCT: difference between VEP P100 latency and PERG P50 latency) was longer (P < 0.01) in POAG than controls and the longer RCT was correlated with the reduced PERG amplitude (r:0.798, P < 0.01). CONCLUSIONS: This suggests that POAG patients have an involvement of the innermost retinal layers and impaired nerve conduction in their visual pathways. PMID- 9352277 TI - Influence of optic and haptic materials on the adherence of Staphylococcus epidermidis to intraocular lenses: a pilot study. AB - To evaluate in vitro the adherence of Staphylococcus epidermidis to intraocular lenses (IOL) of different optic and haptic materials and design, we used a quantitative cultural method. Polymethylmethacrylate (PMMA), PMMA-prolene, polyHEMA, silicone and surface-modified PMMA (wet and dry) implants were tested. Adherence differed significantly in the various groups, with the best performance by all-PMMA IOL. PMID- 9352276 TI - Topical diclofenac sodium compared with prednisolone acetate after phacoemulsification-lens implant surgery. AB - PURPOSE: This study was performed to compare the efficacy, safety and tolerability of diclofenac sodium 0.1% ophthalmic solution with that of prednisolone acetate 1.0% ophthalmic suspension for treatment of inflammation following phacoemulsification and posterior chamber lens implantation. METHODS: One hundred and sixteen patients (diclofenac 57, prednisolone 59) with visually disabling cataract were enrolled in this prospective, randomised, double-masked, parallel group study in two centres. Post-operative patient assessments at day 1, 5-8 and 12-16 included visual acuity, slit-lamp examination, applanation tonometry and subjective evaluation of local tolerance. RESULTS: There was no statistically significant difference between the diclofenac and predisolone groups in the sum of the grades of anterior chamber flare and cells or the degree of conjunctival hyperaemia at any study visit. The overall assessment of local tolerance was similar for both the study medications. There were two (3.4%) possibly drug-related adverse events in the prednisolone group but neither was severe. CONCLUSIONS: Diclofenac sodium 0.01% ophthalmic solution was as effective, safe and well tolerated overall as prednisolone acetate 1.0% ophthalmic suspension. PMID- 9352278 TI - Choice of surgical technique in the management of cataract combined with vitreous surgery. AB - PURPOSE: To compare different methods of lens removal during vitreous surgery. METHODS: We reviewed the data of 37 consecutive eye operations with combined surgery of the lens and vitreous in the Mulheim Eye Hospital between March '93 and September '94. RESULTS: In 14 eyes a pars plana lensectomy was done, in 7 an ECCE (extra capsular cataract extraction), and in 16 phacoemulsification was combined with a regular three-port pars plana vitrectomy. CONCLUSIONS: The choice of procedure was mainly influenced by the hardness of the lens and whether an IOL implant was considered. If no IOL is planned and the lens is soft enough, the best way to remove it is by pars plana lensectomy. If the nucleus seems too hard, phacoemulsification should be performed, because of the risk of releasing the nucleus into the posterior segment. If an IOL is planned, the best method of lens removal is phacoemulsification via a scleral tunnel. In both cases if the nucleus is very hard ECCE should be performed because of the risk of corneal edema. PMID- 9352279 TI - The significance of serum anti-Borrelia antibodies in the diagnostic work-up of uveitis. AB - PURPOSE: To assess the utility of testing uveitis patients for anti-Borrelia antibodies in an area endemic for Lyme borreliosis. METHODS: We examined 161 uveitis patients for serum antibodies to Borrelia burgdorferi by Lyme ELISA. Antibodies were determined in patients with uveitis of unknown etiology and non selectively from patients with an established diagnosis. RESULTS: Concentrations of antibodies to B. burgdorferi were elevated in 26 uveitis patients (16.1%), with elevated IgG in 11 of them (6.8%). In four of these patients Lyme borreliosis was a highly suggestive cause of uveitis because of a history of tick bites, systemic symptoms, response to antibiotic therapy, and/or a positive polymerase chain reaction result. Other causes of uveitis were ruled out. All these patients had vitritis. CONCLUSIONS: Non-selective testing of uveitis patients for Lyme antibodies is not reasonable even in endemic areas. We recommend using the Borrelia antibody test only in cases of uveitis of unknown cause, especially in patients with vitritis or other symptoms of Lyme borreliosis. PMID- 9352281 TI - Optic nerve hypoplasia in fetal alcohol syndrome: an update. AB - Optic nerve hypoplasia was detected in up to one half of a group of Swedish children born to alcoholic mothers. Using an experimental model of pre- and postnatal alcohol exposure in rats fed a liquid diet, reduced optic nerve size from gestational day 21 (294 +/- 26 x 10(2) microns2 vs 502 +/- 16 x 10(2) microns2; n = 6; p < or = 0.001) to later in development was observed as a result of the daily mean blood alcohol levels achieved in dams and their offspring. Altered glial cells and degenerating and atrophic optic axons, myelin sheaths and ganglion cells were frequent in the alcohol-exposed optic nerves. Smaller optic nerve (1.918 +/- 61 x 10(2) microns2 vs 2.195 +/- 40 x 10(2) microns2; n = 4; p < or = 0.001), reduced gaglion cell and axonal densities, and ultrastructural damage to the macroglial cells and myelin sheaths were also detected in the treated group. All these changes remained in the retina and optic nerve of the oldest rats, as a consequence of the long-lasting effects of prenatal alcohol exposure. In summary, alcohol as a major teratogenic agent may induce dysmorphogenesis and irremediable damage to the retina and optic nerve, which frequently manifests itself as hypoplastic optic nerve. PMID- 9352280 TI - Color Doppler ultrasound in ocular Behcet's disease. AB - The purpose of this study was to evaluate the hemodynamic changes occurring in the ophthalmic vasculature of eyes with Behcet's disease in a controlled clinical trial. Both eyes of patients with retinal involvement due to Behcet's disease were consecutively evaluated and were established as having mild or severe retinal vasculitis according to the ophthalmoscopic and fundus fluorescein angiographic findings. One eye from each patient was randomly selected and 25 eyes with mild to moderate and 25 eyes with severe vasculitis were identified. Color Doppler imaging (CDI) was used to quantitate blood flow velocities and vascular resistance in the ophthalmic artery (OA), central retinal artery (CRA) and central retinal vein (CRV) of these patients and those of 25 healthy volunteers. All three groups were age- and sex-matched. In the OA, peak systolic, end diastolic and average flow velocities were significantly higher in patients with Behcet's disease than in the control group (p < 0.05). CRA blood flow velocities of patients with severe retinal involvement were significantly lower than those with mild to moderate vasculitis and control groups and the average vascular resistance was significantly higher than in the control group (p < 0.05). Additionally, the average blood flow velocities in the CRV of patients with severe vasculitis were significantly lower than in mild to moderate vasculitis and control patients. Marked circulatory changes were seen in the ophthalmic vasculature of eyes with Behcet's disease. Although larger studies are required to define the true sensitivity and specificity of this technique, these initial results suggest that CDI could play a major part in the assessment of patients with ocular Behcet's disease. PMID- 9352282 TI - Acquired bilateral superior oblique palsy: a localising sign in the dorsal midbrain. AB - Bilateral superior oblique palsy is an uncommon ocular motility problem, the commonest cause being closed head trauma. Two cases, both adults, are presented in whom bilateral superior oblique palsy occurred as a result of neoplastic infiltration of the dorsal midbrain in the region of the anterior medullary velum. In the absence of a history of head trauma, the presence of an acquired bilateral superior oblique palsy is a definite sign of a single lesion in the region of the decussation of the trochlear nerves and appropriate imaging is indicated. PMID- 9352283 TI - Relationship between retinal lesions and axial length, age and sex in high myopia. AB - We analysed the relationship between central and peripheral retinal lesions and axial length (AL), patient's age and sex with myopia greater than 6 diopters. A total of 212 eyes of 109 patients with high myopia underwent detailed funduscopy and A-scan ultrasonography. AL was measured, and central and peripheral retinal lesions were noted. Results were analysed using Student's t-test. Sixty-one patients (118 eyes) were female and 48 (94 eyes) male. Mean age was 31.00 +/- 13.67 years, and mean AL was 28.31 +/- 2.02 mm. Chorioretinal atrophy, Fuchs' spot, posterior staphyloma and posterior vitreous detachment increased significantly with AL and age. Fuchs' spot was more common in females. White without-pressure (WWP) was inversely correlated with AL and age, and was more common in males. The high frequency of WWP in younger patients and moderate AL suggests that these lesions result from vitreoretinal tractions. Lattice degeneration was also a frequent finding in high myopia, and tended to increase with AL and age, though without reaching statistical significance. PMID- 9352284 TI - Vitreoretinal surgery in diabetic patients on hemodialysis. AB - The present paper reports our first results after pars plana vitrectomy in patients with diabetic retinopathy and hemodialysis with a follow-up of 6 to 24 months. Between January 1992 and October 1994 we performed vitreoretinal surgery with silicone oil tamponade in nine eyes of seven patients with diabetic nephropathy on hemodialysis. All patients had had type I diabetes for 19-32 years. Over the observation period the retina was completely attached in eight eyes. Final visual acuity of 0.1-0.7 was attained in four eyes, 0.06 two, hand movements in one eye. Two eyes had no useful final visual acuity because of redetachment of the retina or secondary glaucoma with rubeosis iridis. The small number of complications shows that pars plana vitrectomy can be done in diabetic patients with nephropathy on hemodialysis. This significantly improves their quality of life. PMID- 9352286 TI - Necrotizing choroiditis-retinitis as presenting symptom of disseminated aspergillosis after lung transplantation. AB - BACKGROUND: Endogenous endophthalmitis due to Aspergillus is rare affecting the severely immunosuppressed population, in particular recipients of heart and lung transplants. Ocular involvement of aspergillosis has always been observed late in the course of the disease. SUBJECT: A young woman noted blurred vision in one eye three weeks after lung transplantation. At this stage, no systemic manifestations of fungal infection were detected and the ocular findings were attributed to viral infection. RESULTS: Twenty-four hours after the original ocular complaint, an aggressive endophthalmitis developed in the left eye. The possibility of fungal endophthalmitis was raised. Within 48 hours of her first ocular complaint the patient died. Cultures from a vitreous tap and from autopsy ocular specimens were positive for Aspergillus fumigatus. CONCLUSIONS: Aspergillus endopthalmitis may occur in patients undergoing lung transplantation despite antifungal therapy. Increased awareness of this unusual entity may be life and vision saving in these patients. PMID- 9352285 TI - 1-Octadecene as a solvent for ferrofluids for intraocular use. AB - PURPOSE: 1-Octadecene is a hydrocarbon with one double bond in its structure that could serve as a solvent for ferrofluids. The aim of this pilot study was to obtain preliminary information on intraocular tolerance to 1-octadecene. METHODS: Vitreous compression with perfluoropropane gas was achieved in 20 eyes of albino rabbits. Four days after gas injection a fluid-gas exchange was undertaken. Sixteen eyes received 1-octadecene. Four eyes received balanced salt solution. Eyes were obtained at 3, 7, 14 and 30 days. The samples were fixed in 10% buffered formalin, processed in paraffin and sections were stained with hematoxylin and eosin. RESULTS: Emulsification of the oil bubble was observed in 31.25% of the cases by the fifth day; light microscopy showed normal retinal architecture in all the eyes and epiretinal and vitreous macrophages in 50% of the eyes. CONCLUSIONS: 1-Octadecene does not appear to have any retinal cytotoxic effect but elicits an inflammatory response in the vitreous activity. PMID- 9352287 TI - Central serous chorioretinopathy complicating systemic corticosteroid therapy. AB - PURPOSE: To present evidence that systemic corticosteroid therapy may cause central serous chorioretinopathy. METHODS: A 20-year-old male with idiopathic thrombocytopenic purpura was examined during systemic treatment with corticosteroids (100 mg daily). RESULTS: The patient had central serous chorioretinopathy. Spontaneous recovery accompanied discontinuation of the steroid treatment. CONCLUSIONS: This case provides further evidence that cortisol may play a role in the development of central serous chorioretinopathy. PMID- 9352288 TI - Orbital metastasis of renal cell carcinoma masquerading as Amaurosis fugax. AB - Renal cell carcinoma (RCC) is the most common malignancy involving the kidney. Only rarely does it metastasize to the eye and orbit, sometimes mimicking other lesions. A 70-year-old woman was referred from neurology because of a right orbital lesion, six months after the start of a neurological investigation because of amaurosis fugax. Six months earlier she had complained of transient visual disturbances in her right eye. After excluding cardiovascular abnormalities and coagulopathies as the source of her complaints, she was diagnosed as having a right senile ptosis. A computed tomography scan, done to complete the workout, detected a right orbital mass. The patient was referred to the oculoplastic unit. A biopsy and then a lateral orbitotomy were performed. Histopathological examination proved it to be a metastatic renal cell carcinoma, seven years after the primary tumor had been diagnosed and treated by nephrectomy. The characteristics of metastatic renal cell carcinoma are discussed, in view of the rarity of metastasis to the eye and, in particular, to the orbit, and its tendency to masquerade as other lesions or symptoms. In this case it presented as amaurosis fugax before other signs appeared. PMID- 9352289 TI - Executive control and the comprehension of medical information by elderly retirees. AB - This study examined the independent contributions of executive control function, general cognition, age, education, and medication usage to the comprehension of medical information. Randomly selected elderly retirees (N = 105) more than 70 years of age completed the Executive Interview (EXIT25), the Mini-Mental State Exam (MMSE), and the Hopkins Competency Assessment Test (HCAT). Cognitive measures were stronger predictors of HCAT scores than age, education, or number of prescribed medications. A discriminant model based on EXIT25 and MMSE scores correctly classified 91% of subjects relative to their HCAT scores. It was concluded that executive impairment is strongly associated with impaired comprehension of medical information. As many as 88% of probable Alzheimer's disease patients, 69% of institutionalized elderly retirees, and 49% of noninstitutionalized retirees may be impaired in their ability to comprehend medical information, even when it has been presented well below their educational level. PMID- 9352290 TI - Metacognition and medication adherence: how do older adults remember? AB - Fifty-one older adults (M age = 75.9 years, SD = 6.9) reported their use of memory strategies for taking of medication using the Prospective Memory for Medication Questionnaire. Older adults used internal strategies more often when the domain was restricted to medication taking but used external strategies more often when queried across a variety of everyday situations. Surprisingly, the hypothesis that medical factors would be the primary determinants of older adults' reports of memory strategy use and perceived adherence was not supported. Metamemorial variables of non-domain-specific memory self-efficacy and memory anxiety in everyday life were significant predictors of strategy use and perceived adherence over and above variables related to the domain of health. PMID- 9352291 TI - Aging and mind wandering: reduced inhibition in older adults? AB - Hasher and Zacks (1988) theorize that aging disrupts the efficient operation of an inhibitory mechanism that, when functioning normally, is thought to suppress information irrelevant to one's cognitive goals. Problems with this inhibitory mechanism should produce increased mind wandering, and the present experiment examined this possibility using a performance-based measure of mind wandering. Younger and older participants were presented with a long list of words and were occasionally stopped (at unpredictable intervals) and asked to recall the most recently presented items. Mind wandering was inferred by conditionalizing recall on these unpredictable trials with recall on short, predictable trials (in which, presumably, participants were able to maintain full attention to the recall task). Whereas mind wandering was shown to be higher on longer trials than shorter ones, there was no evidence of age differences in mind wandering. PMID- 9352292 TI - Age and forgetfulness: absolute versus comparison decisions about capability. AB - Perceivers were assigned to one of two decision conditions. In an absolute decision condition, perceivers rated how likely they would be to allow a young or old highly forgetful, slightly forgetful, or nonforgetful target to perform a challenging task. In a comparison decision condition, perceivers rated two targets, one young and one old, who had a similar level of forgetfulness. Separate Decision Type x Target Forgetfulness analyses of variance were conducted on ratings of the two target age groups. Young targets received higher ratings in the comparison than in the absolute condition, whereas old targets were rated the same in the two conditions. There was some preference for young targets in a comparison situation, but it was concluded that forgetfulness was a more important factor than age in perceivers' ratings. PMID- 9352293 TI - Reach profiles of men and women 65 to 89 years of age. AB - Reach and anthropometric data were collected on a heterogeneous group of subjects 65 to 89 years of age. The purpose of this pilot study was to determine whether the process of aging has an effect on the ability to reach. Analysis of the data collected showed that age-related changes had an effect on reaching in the vertical plane, especially for the male subjects. The vertical reach fingertip and grip (VRT and VRG) measurements for men 85 to 89 years old were significantly lower than those for all other age groups, indicating that reaching abilities decrease between the ages of 80 and 85 years. The results for women were not as conclusive, but additional analysis showed a decline in VRT and VRG in terms of stature. Horizontal reach measurements showed no age-related effect in the analysis. An attempt was made to compare reach data collected here with those collected in previous studies. However, comparisons of the reach data were limited as a result of differences in reaching activities and measuring techniques in other studies. Anthropometric measurements of this older cohort were compared with those of a younger population. It was noted that older subjects had smaller anthropometric measurements than their younger cohorts. Differences in body size and reach, whether statistically significant or not, need to be considered when designing living environments and appliances for the aged. PMID- 9352294 TI - Sulfated polysaccharides extracted from sea algae as potential antiviral drugs. AB - The inhibitory effects of polyanionic substances on the replication of herpes simplex virus (HSV) and other viruses were reported almost four decades ago. However, these observations did not generate much interest, because the antiviral action of the compounds was considered to be largely nonspecific. Shortly after the identification of human immunodeficiency virus (HIV) as the causative agent of the acquired immune deficiency syndrome (AIDS) in 1984, heparin and other sulfated polysaccharides were found to be potent and selective inhibitors of HIV 1 replication in cell culture. Since 1988, the activity spectrum of the sulfated polysaccharides has been shown to extend to various enveloped viruses, including viruses that emerge as opportunistic pathogens (e.g., herpes simplex virus [HSV] and cytomegalovirus [CMV]) in immunosuppressed (e.g., AIDS) patients. As potential anti-HIV drug candidates, sulfated polysaccharides offer a number of promising features. They are able to block HIV replication in cell culture at concentrations as low as 0.1 to 0.01 microgram ml-1 without toxicity to the host cells at concentrations up to 2.5 mg ml-1. We noted that some polysulfates show a differential inhibitory activity against different HIV strains, suggesting that marked differences exist in the target molecules with which polysulfates interact. They not only inhibit the cytopathic effect of HIV, but also prevent HIV-induced syncytium (giant cell) formation. Furthermore, experiments carried out with dextran sulfate samples of increasing molecular weight and with sulfated cyclodextrins of different degrees of sulfation have shown that antiviral activity increases with increasing molecular weight and degree of sulfation. A sugar backbone is not strictly needed for the anti-HIV activity of polysulfates because sulfated polymers composed of a carbon-carbon backbone have also proved to be highly efficient anti-HIV agents in vitro. Other, yet to be defined, structural features may also play an important role. Sulfated polysaccharides may act synergistically with other anti-HIV drugs (e.g., azidothymidine [AZT]). They are known to lead very slowly to virus-drug resistance development and they show activity against HIV mutants that have become resistant to reverse transcriptase inhibitors, such as AZT, tetrahydro-imidazo [4,5,l-jk] [1,4]-benzodiazepin-2(1H) thione (TIBO) and others. From studies on their mechanism of action we concluded that polysulfates exert their anti-HIV activity by shielding off the positively charged sites in the V3 loop of the viral envelope glycoprotein (gp120). The V3 loop is necessary for virus attachment to cell surface heparan sulfate, a primary binding site, before more specific binding occurs to the CD4 receptor of CD4+ cells. This general mechanism also explains the broad antiviral activity of polysulfates against enveloped viruses. Variations in the viral envelope glycoprotein region may result in differences in the susceptibility of different enveloped viruses to compounds that interact with their envelope glycoproteins. The efficacy of polysulfates in the therapy and/or prophylaxis of retroviral infections and opportunistic infections remains to be demonstrated both in animal models and humans. It is important to consider not only treatment of patients who are already infected with HIV, but also prophylaxis and protection from HIV and/or other virus infections. Because (i) sexual transmission is responsible for the large majority of HIV infections worldwide; (ii) this transmission is mostly mediated via mononuclear cells that infect epithelial cells of the genital tract; and because (iii) polysulfates effectively inhibit cell-cell adhesion, polysulfates may be considered as potentially effective in a vaginal formulation to protect against HIV infection. PMID- 9352295 TI - The role of DNA damage in the cytotoxic response to hydrogen peroxide/histidine. AB - 1. Histidine enhances the cytotoxic and clastogenic effects of hydrogen peroxide. In this review, we will focus on two lesions that are generated in the presence of histidine in oxidatively injured cells--namely, DNA single- and double-strand breaks (SSBs and DSBs). 2. Hydrogen peroxide is a potent inducer of DNA SSBs, and histidine modulates the formation of these lesions. This effect has been extensively characterized with the use of purified DNA, and the results obtained have demonstrated that, upon exposure to low or high concentrations of H2O2, histidine reduces or enhances the formation of DNA SSBs, respectively. The protective effect has been ascribed to iron chelation, whereas the enhancing effect is probably the consequence of the formation of a histidine/iron/DNA complex. 3. In cultured cells, histidine potentiates the formation of H2O2 induced DNA SSBs but these lesions are efficiently repaired and do not appear to mediate the cytotoxic response. 4. In the presence of micromolar levels of histidine, H2O2 also induces DNA DSBs, a type of lesion that is not generated by the oxidant alone. The experimental evidence that has been thus far collected would suggest that these DNA DSBs are toxic and are indeed the cause of cell death induced by the cocktail H2O2/histidine. PMID- 9352296 TI - Effects of adenosine and isoprenaline in left atria from both neonatal and middle aged noninsulin-dependent diabetic rat models. AB - 1. This study examined the ability of atria from neonatal and middle-aged noninsulin-dependent diabetic rat models to respond to both adenosine and isoprenaline. 2. Cumulative additions of adenosine (1-1000 microM) produced concentration-dependent decreases in the force of contraction of rat atria that were unchanged in neonatal diabetic animals. Although direct inotropic responses to adenosine were unchanged, atria from neonatal diabetic animals exhibited an increase in maximum response to adenosine-induced antiadrenergic effect. 3. Atria from middle-aged noninsulin-dependent diabetic rats exhibited a supersensitivity to the direct inotropic effect of adenosine compared with atria from age-matched control rats. The middle-aged, noninsulin-dependent diabetic state did not alter the maximum response of atria to adenosine-induced antiadrenergic effect. 4. A comparison was made between middle-aged (10-month-old) controls and young (4 month-old) controls. Atria from middle-aged control animals exhibited a lower sensitivity and responsiveness to the direct inotropic effect of adenosine compared with those from young controls. 5. Cumulative additions of isoprenaline (10(-9)-10(-6) M) produced concentration-dependent increases in inotropy that were unchanged in atria from either neonatal or middle-aged noninsulin-dependent diabetic rats. 6. These results show that neonatal and middle-aged noninsulin dependent diabetes and age-related factors lead to significant changes in atrial reactivity to the adenosine-induced stimulation in the absence and presence of isoprenaline. However; isoprenaline-induced positive inotropic response cannot change in each diabetic heart to an apparent extent. PMID- 9352297 TI - Modulation by APGW-amide, an Achatina endogenous inhibitory tetrapeptide, of currents induced by neuroactive compounds on Achatina neurons: amines and amino acids. AB - 1. Modulatory effects of APGW-amide (Ala-Pro-Gly-Trp-NH2), proposed as an inhibitory neurotransmitter of Achatina neurons, perfused at 3 x 10(-6) M on the currents induced by small-molecule putative neurotransmitters were examined by using Achatina giant neuron types, v-RCDN (ventral-right cerebral distinct neuron), TAN (tonically autoactive neuron) and RAPN (right anterior pallial nerve neuron), under voltage clamp. These putative neurotransmitters were ejected locally to the neuron by brief pneumatic pressure. 2. Outward current (Iout) induced by erythro-beta-hydroxy-L-glutamic acid (erythro-L-BHGA) on v-RCDN, which was probably K+ dependent, was enhanced with membrane conductance (g) increase under APGW-amide. From dose (pressure duration)-response curves of erythro-L-BHGA measured in physiological solution (control curve) and with APGW-amide (drug curve), ED50 values of the two curves were nearly comparable, whereas Emax of the drug curve was significantly larger than that of the other. From a Lineweaver Burk plot of these data, the cross point of the control line and the drug line was on the abscissa. 3. K(+)-dependent Iout caused by dopamine (DA) on v-RCDN was inhibited with a g increase by APGW-amide. The inhibition of this current caused by APGW-amide was mainly in a noncompetitive and partly uncompetitive manner. 4. 5-Hydroxytryptamine (5-HT) produced an inward current (Iin) with two (fast and slow) components on TAN, which was probably Na+ dependent. The fast component of the Iin was inhibited by APGW-amide. The inhibition was mainly in a noncompetitive manner. 5. The currents induced by acetylcholine, gamma aminobutyric acid and L-glutamic acid on Achatina neuron types were not affected by APGW-amide. 6. The inhibitory effects of APGW-amide on the Iin (fast component) induced by 5-HT were nearly equipotent or a bit stronger than those on the Iout caused by DA. 7. The g increase produced by APGW-amide would be a cause for inhibiting the Iout induced by DA. In addition, we consider that APGW-amide affects intracellular signal transduction systems or ionic channels, thus modulating these currents. PMID- 9352298 TI - Modulation by APGW-amide, an Achatina endogenous inhibitory tetrapeptide, of currents induced by neuroactive compounds on Achatina neurons: peptides. AB - 1. Modulatory effects of APGW-amide (Ala-Pro-Gly-Trp-NH2), proposed as an inhibitory neurotransmitter of Achatina neurons, perfused at 3 x 10(-6) M on the currents induced by neuroactive peptides, ejected by brief pressure, were examined by using Achatina giant neuron types, v-RCDN (ventral-right cerebral distinct neuron) and PON (periodically oscillating neuron), under voltage clamp. 2. Outward current (Iout) caused by FMRFamide (Phe-Met-Arg-Phe-NH2) on v-RCDN, which was probably K+ dependent, was inhibited with membrane conductance (g) increase by APGW-amide. From the dose (pressure duration)-response curves of FMRFamide and a Lineweaver-Burk plot of these data, the inhibition caused by APGW amide was mainly in an uncompetitive manner. 3. Iout caused by APGW-amide on v RCDN, which was probably K+ dependent, was inhibited with g increase by APGW amide. The inhibition caused by APGW-amide was partly in a competitive manner and partly in a noncompetitive manner. 4. Iout caused by [Ser2]-Mytilus inhibitory peptide, [Ser2]-MIP (Gly-Ser-Pro-Met-Phe-Val-NH2) on v-RCDN, which was probably K+ dependent, was inhibited with g increase by APGW-amide. Because the modulation of this current was not so marked, a dose-response study of this compound was not carried out. Iin induced by oxytocin on PON was not affected by APGW-amide. 5. From the dose-response curves of APGW-amide, perfused consecutively, the inhibitory effects of APGW-amide on the Iout caused by APGW-amide were stronger than those on the Iout caused by FMRFamide. 6. The inhibition of the APGW-amide induced Iout on v-RCDN by APGW-amide was partly due to the competition in the receptor sites and partly to the g increase. The inhibition by APGW-amide on the Iout induced by FMRFamide and [Ser2]-MIP would be partly due to the g increase. In addition, we consider that APGW-amide affects intracellular signal transduction systems or ionic channels, thus modulating these currents. 7. The currents modulated by APGW-amide were different from those modulated by achatin 1, another Achatina endogenous neuroexcitatory peptide. We consider that the mechanisms underlying the modulatory effects of APGW-amide are different from those of achatin-I. PMID- 9352299 TI - Effects of several class I antiarrhythmic drugs on isolated rat aortic vascular smooth muscle. AB - 1. The vasorelaxant effects of seven NA+ channel blockers (i.e., class I antiarrhythmic agents), quinidine, disopyramide, imipramine, lidocaine, mexiletine, flecainide, and desipramine, were investigated in isolated endothelium-denuded rat aorta. 2. All drugs induced a concentration-dependent relaxation in aorta precontracted with either 80 mM KCl or 10(-5) M noradrenaline and, with the exception of mexiletine, they were more potent in inhibiting KCl induced contractions. 3. The degree of inhibition of high KCl-induced contractions produced by quinidine and desipramine increased with the time of depolarization. Furthermore, the inhibitory effect of quinidine also increased in aorta preincubated in 40 mM KCl, whereas the inhibitory effects of other antiarrhythmics were almost similar in 5 or 40 mM KCl solution. 4. In conclusion, all these class I antiarrhythmic drugs inhibited Ca2+ entry through voltage- and receptor-gated channels as well as Ca2+ release from intracellular stores. As a consequence, they decrease the availability of intracellular free Ca2+ required for vascular smooth muscle contraction. PMID- 9352300 TI - Antianginal effects of YM430, a novel calcium entry-blocking and beta adrenoceptor-blocking agent in several experimental angina models. AB - 1. We evaluated the antianginal effects of YM430 in several experimental models in vitro and in vivo. 2. In isolated dog coronary artery, YM430 (10(-8)-10(-6) M) inhibited 3,4-diaminopyridine-induced rhythmic contractions with an IC50 value of 59.2 nM. 3. In anesthetized rats, YM430 (10-100 mg/kg PO) inhibited arginine vasopressin-induced ST-segment depression with an IC50 value of 36.6 mg/kg PO. 4. In anesthetized dogs, YM430 (0.3 mg/kg IV) significantly inhibited ST-segment elevation induced by coronary artery occlusion. 5. These findings suggest that YM430 may be of value in the treatment of various types of angina pectoris such as variant and stable angina. PMID- 9352301 TI - Changes in the activities of antioxidant enzymes of the lymphoid organs of 21-day pregnant rats due to administration of fish oil by gavage. AB - 1. The effect of fish oil administration by gavage (0.4% body weight) on activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) and on content of thiobarbituric acid reactive substances (TBARs) of the lymphoid organs [thymus, spleen and mesenteric lymph nodes (MLN)] and liver was investigated in 21-day pregnant rats. The results were compared with those obtained by administration of soybean oil, cocoa butter and coconut oil. 2. Oil administration did not have any significant effect on antioxidant enzyme activities of the liver, whereas marked changes were found in the lymphoid organs. The MLN presented the most pronounced changes: SOD and catalase activities were increased by the four oils; GSH-Px activity was raised by soybean and fish oils; coconut oil reduced the activity of the three antioxidant enzymes in this organ. 3. Fish oil given by gavage does affect the antioxidant capacity of the lymphoid organs; however, similar effect was also observed for cocoa butter and soybean oil. These changes in the antioxidant enzyme activities were able to prevent the lipid peroxidation process in the lymphoid organs. PMID- 9352302 TI - Influence of extracellular H+ and Ca2+ on Ro 22-9194-induced block of sodium current in cardiac myocytes. AB - 1. Ro 22-9194 reduced the Na current in ventricular myocytes in either a tonic block or phasic block manner. 2. Ro 22-9194 had a higher affinity to the inactivated state (Kdi = 10.3 microM) than to the rested state (Kdrest = 180 microM). 3. Extracellular acidification enhanced the tonic block but reduced the phasic block. 4. Elevation of extracellular Ca2+ inhibited the enhancing effects of extracellular acidification. 5. These findings suggest that Ro 22-9194 strongly inhibits Na+ channels of the ventricular myocytes of the diseased hearts, characterized by the depolarized cell membranes and by acid conditions. PMID- 9352303 TI - Changes in isoprenaline-induced endothelium-dependent and -independent relaxations of aorta in long-term STZ-diabetic rats: reversal effect of dietary vitamin E. AB - 1. The present study concerns in vitro isoprenaline (ISO)-induced relaxation of aortic rings of long-term streptozotocin (STZ)-diabetic and nondiabetic rats, both with and without dietary vitamin E supplementation. 2. Incubation with propranolol, NG-nitro-L-arginine methyl ester and methylene blue, as well as absence of endothelium, all negatively affect the ISO-induced relaxations. 3. Thiobarbituric acid reactivity levels used as an index of lipid peroxidation are elevated in the aorta by diabetes. Four months of STZ-diabetes results in a significant increase in the ISO-induced relaxations together with endothelial dysfunction in the rat aorta. Diabetes also causes the loss of vascular integrity. 4. Dietary vitamin E supplementation during the last 2 months of diabetes allows normalization of the levels of lipid peroxides. This vitamin also completely reverses the increased sensitivity (pD2 value) of the aorta to ISO, whereas the maximum ISO-induced relaxations are partially restored after the treatment in diabetic rats. 5. The results suggest that ISO-induced relaxation in the aorta partially depends on the intact endothelium and that the endothelium dependent relaxant effect of ISO is mediated by endothelium-derived relaxing factor. Results also indicate that abnormal vascular reactivity and structure of the diabetic rat aorta may be related to the increased lipid peroxidation. In conclusion, vitamin E can protect the arterial wall from oxidative stress-induced injury associated with chronic STZ-diabetes and allows normalization of the response to ISO and the structure of the aorta in diabetic rats. PMID- 9352304 TI - Altered heme pathway regulation and drug metabolizing enzyme system in a mouse model of hepatocarcinogenesis: effect of veronal. AB - 1. Male CF 1 mice were fed p-dimethylaminoazobenzene (DAB) for 35 days and received 5,5-diethylbarbituric acid, before or after DAB treatment, with the purpose of investigating whether the onset of the preinitiation stage of carcinogenesis alters the natural regulatory mechanism of the heme pathway. 2. Changes detected in drug metabolizing enzymes are likely to be the consequence of a primary deregulation mechanism of heme metabolism, shown by an increase in delta-aminolevulinic acid synthetase activity and a decrease in microsomal heme oxygenase, which would finally lead to a great enhancement of cytochrome P450 levels. 3. The alterations found here would give rise to a pattern distinctive to that usually observed in the so-called resistant hepatocyte. PMID- 9352305 TI - Multiple forms of AMPA-type glutamate receptor mRNA phenotypes in goldfish retina and tectum. AB - 1. A goldfish AMPA-type glutamate receptor cDNA (GFGR49) was cloned from a goldfish retinal cDNA library. 2. The GFGR49 clone is an immature product of pre mRNA, possessing the C-flip exon and flanking introns. 3. RNase protection assays revealed multiple mRNAs that contain the C-flip exon. In addition, these assays indicated differential processing of these RNAs between retina and brain. 4. Finally, RNase protection assays employing probes spanning the 3'-coding region of GFGR49 revealed at least two different mRNA phenotypes. 5. Comparison with the sequence of genomic DNA, which was obtained by polymerase chain reaction amplification, suggests that such multiple mRNA phenotypes are attributed to the use of alternative intron-exon splicing junctions and to RNA editing. PMID- 9352307 TI - Effect of locust poison on neuromembrane functional activity. AB - 1. The effect of locust poison (LP) on membrane excitability, chemosensitivity and Na-K pump activity was studied. 2. LP in a concentration of 10(-4) mg/ml has a potential independent blocking effect on neuromembrane excitability. 3. LP has an activation effect on Na-K pump-induced membrane hyperpolarization. 4. In a concentration higher than 10(-4) mg/ml, LP leads to a decrease in the acetylcholine-induced current and to an increase in this current after the washout of LP from the medium. PMID- 9352306 TI - Effects of sweetening agents on morphine-induced analgesia in mice by formalin test. AB - 1. There is evidence that sweet-tasting substances such as sucrose and saccharin can interact with endogenous opioid systems. Further evidence showed that feeding mice different concentrations of sucrose and saccharin alter the latency in the tail-flick test. 2. In the current study, the effects of a 12-day regimen of different sweetening agents [sucrose (32%), saccharin (0.08%) and aspartame (0.16%)] on morphine-induced analgesia with the formalin test were investigated. 3. Male albino mice (20-27 g) were used for the experiments. Animals were given 12 days to adapt to dietary conditions. Animals were first given saline or morphine subcutaneously (1.5, 3.0, 6.0, or 9.0 mg/kg) 30 min before the observation period. The recording of the early phase started immediately and lasted for 10 min. The recording of the late response started 20 min after formalin injection and lasted for 10 min. Statistical analysis was performed by using analysis of variance followed by Newman-Keuls test, and P < or = 0.05 was considered significant. 4. Sucrose and aspartame increased morphine analgesia in the early phase, but saccharin had no effect on the early phase. On the other hand, saccharin and sucrose decreased the effect of morphine in the late phase, but aspartame increased the effect of morphine-induced analgesia. 5. In conclusion, the present data provide further evidence for an important role for dietary variables in determining the effects of exogenous opioids on pain sensitivity. PMID- 9352308 TI - Effects of phorbol-12,13-dibutyrate and protein kinase C inhibitors on Mn(2+) dependent norepinephrine-induced contractions involving increase in Mn2+ sensitivity in Ca(2+)-depleted vas deferens of the guinea pig. AB - 1. In Ca(2+)-depleted Mn(2+)-loaded vas deferens from the guinea pig (Mn-loaded preparations), norepinephrine (NE) induced a tonic contraction dose dependently without extracellular Ca2+ and Mn2+. 2. In the beta-escin skinned vas deferens, Mn2+ as well as Ca2+ induced contractions. Guanosine triphosphate and NE increased the sensitivity of contractile mechanisms to these divalent cations. 3. Phorbol-12,13-dibutyrate (PDBu) did not induce contractions in normal (Mn(2+) unloaded Ca(2+)-contained) preparations, whereas it induced slow sustained contractions dose dependently in Mn-loaded preparations. Although cumulative applications of PDBu desensitized the preparations to this phorbol ester, the desensitization did not affect Mn(2+)-dependent NE-induced contractions. PDBu did not affect the dose-response relation of NE in Mn-loaded preparations. 4. Staurosporine (10-100 nM) preferentially inhibited NE-induced contractions in normal and in Mn-loaded preparations to that induced by K+ in normal preparations. However, bisindolylmaleimide I (1 microM) did not inhibit NE induced contractions in normal and Mn-loaded preparations but abolished PDBu induced contractions. 5. These results suggest that the NE-induced increase in the Mn2+ sensitivity of contractile mechanisms contributes to Mn(2+)-dependent NE induced contractions, which may not involve the activation of protein kinase C. PMID- 9352310 TI - Effects of erythromycin on chemoattractant-activated human polymorphonuclear leukocytes. AB - 1. Erythromycin (2-100 micrograms ml-1) produced a concentration-related inhibition of superoxide generation and elastase release induced by in vitro exposure of human polymorphonuclear leukocytes (PMNs) to the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (FMLP; 30 nM). 2. By contrast, erythromycin (100 micrograms ml-1) did not alter the leukotriene B4 production elicited by FMLP (30 nM; in the presence of thimerosal 20 microM) or the intracellular calcium changes promoted by FMLP (30 nM; in the absence or presence of thimerosal 20 microM). 3. These results indicate that by reducing chemoattractant-triggered release of oxidative and proteolytic mediators from human PMNs, erythromycin may have clinically useful antiinflammatory effects. PMID- 9352309 TI - Purinergic modulation of rat urinary bladder detrusor smooth muscle. AB - 1. Rat detrusor muscle was responsive to both ATP and adenosine; ATP elicited an excitatory response, whereas adenosine had an inhibitory effect. 2. ATP and adenosine had an inhibitory modulatory action on responses to acetylcholine, potassium depolarization and field stimulation. 3. Quinidine inhibited the ATP response and blocked the inhibitory effect of ATP on acetylcholine, potassium depolarization and field-stimulation responses. The effect of adenosine remained unaltered in the presence of quinidine. 4. Caffeine and theophylline blocked the adenosine inhibition of responses to field stimulation. 5. It is concluded that excitatory P2-type purinoreceptors mediated by ATP and inhibitory P1-type purinoreceptors mediated by adenosine exist in rat urinary bladder detrusor smooth muscle and that both ATP and adenosine exhibit a modulatory action on detrusor muscle agonist-induced responses. PMID- 9352311 TI - Endothelin receptor-mediated Ca2+ mobilization and contraction in bovine oviductal arteries: comparison with noradrenaline and potassium. AB - 1. The effects of endothelin-1 (ET-1) were studied in bovine oviductal arteries and compared to those of noradrenaline (NA) and high K+ (K+). The influence of endothelium, the receptor subtypes involved, and the mechanisms of Ca2+ mobilization were assessed. 2. ET-1 (0.1-300 nM) induced concentration-dependent contractions with a potency of 10(3) and 10(2) times higher than NA (0.1 microM 0.1 mM) and K+ (9.5-119 mM), respectively. Removal of endothelium or NG-nitro-L arginine (L-NOARG, 0.1 mM) pretreatment did not affect responses to either ET-1 or K+, whereas the NA response was significantly increased. Indomethacin (1 microM) had no effect on either of these agonists. 3. The rank order of potency for the ET isopeptides was: ET-1 = ET-2 > ET-3. The ETA receptor-selective agonist, sarafotoxin 6c (S6c), had no effect. The ETA receptor-selective antagonist, BQ-123, showed a competitive antagonism on the ET-1 response (pA2 value of 6.58 +/- 0.01), whereas contractions to ET-3 were completely abolished by BQ-123 at 0.1 microM. 4. Concentration-response curves to both ET-1 and NA were shifted to the right and their maximum response reduced to approximately 56% and 65% of controls, respectively, under 30 min of incubation in Ca(2+)-free solution, whereas responses to K+ were almost abolished by this treatment. Contractions to both NA (30 microM) and ET-1 (30 nM) were maximally inhibited after 10 min of extracellular Ca2+ deprivation. 5. Contractions to ET-1 were more potently inhibited by nickel (Ni2+, 0.3 mM), whereas nifedipine (1 microM) and cadmium (Cd2+, 0.1 mM) induced only a slight effect. In contrast, opposite effects were found for both NA and K+. 6. Treatment with ryanodine (100 microM) and caffeine (10 mM) in Ca(2+)-free solution reduced the tension measured 5 min after NA (30 microM) and ET-1 (30 nM) addition, but the sustained response (tension at 25 min) remained unaffected. 7. Calphostin C (1 microM), a specific protein kinase C (PKC) inhibitor, reduced the maximum contractile response to ET 1 by about 50% without significantly affecting its pD2 value. 8. These results suggest that ET-1 acts in bovine oviductal arteries by directly activating a homogenous population of ETA receptors in smooth muscle, without endothelial modulation. Several Ca2+ activation mechanisms seem to be involved in the contractile action of the peptide, including: (1) extracellular Ca2+ entrance through Ni(2+)-sensitive and L-type Ca2+ channels; (2) intracellular Ca2+ release from a ryanodine-sensitive Ca2+ store; and (3) sensitization of the contractile machinery to Ca2+ via PKC. PMID- 9352312 TI - Effects of allocryptopine, an alkaloid isolated from Glaucium arabicum on rat isolated ileum and urinary bladder. AB - 1. The alkaloid, allocryptopine, was isolated from the chloroform extract of Glaucium arabicum. 2. The effect of allocryptopine on urinary bladder and ileal smooth muscles was investigated in this study. 3. Allocryptopine, in concentrations from 1 x 10(-5) to 3 x 10(-3) M caused a concentration-dependent contraction of rat isolated urinary bladder and a concentration-dependent relaxation of rat ileal smooth muscles. 4. Theophylline (10(-5) M) shifted to the left the allocryptopine concentration-effect curve on ileum and increased the maximum inhibitory effect of allocryptopine. 5. Methylene blue (10(-3) M) had no significant effect on the concentration-effect curve of allocryptopine of the ileum. 6. Phentolamine (10(-6) M) shifted to the right the allocryptopine concentration-effect curve of urinary bladder. 7. These observations suggest that allocryptopine induces a relaxing effect on the ileum by inhibiting phosphodiesterase enzyme, and thus elevating cellular cAMP and its contractile effect on the urinary bladder by affecting alpha-adrenergic receptors in this tissue. PMID- 9352313 TI - Ouabain-sensitive K(+)-dependent outward current caused by threo-beta-hydroxy-L glutamic acid on a snail neuron. AB - 1. An analog of L-glutamic acid, threo-beta-hydroxy-L-glutamic acid (threo-L BHGA), was applied locally to the giant neuron of an Achatina snail by pneumatic brief pressure ejection and induced an outward current (Iout) on the ventral-left cerebral distinct neurone (v-LCDN). The present study aimed to elucidate the ionic mechanisms of the Iout caused by threo-L-BHGA (ItL-BHGA) of v-LCDN and the effects of ouabain on this current under voltage clamp. 2. The reversal potentials of ItL-BHGA (EtL-BHGA) of v-LCDN in varied K+o were fitted to the Nernst equation as ItL-BHGA = IK (K+ current) and were almost unchanged in Cl-o free and Na+o-reduced (20% of normal) states. The ItL-BHGA is due to the increase in permeability of the neuromembrane to K+(K(+)-dependent) and is neither Na(+)- nor Cl-(-)-dependent. K(+)-channel blockers, a mixture of tetraethyl-ammonium (TEA) and 4-amino-pyridine (4-AP), blocked ItL-BHGA mainly in a noncompetitive and partly in an uncompetitive manner. 3. Unexpectedly, ItL-BHGA of v-LCDN was almost abolished in the Na+o-free state and significantly reduced in the Cl-o free state. However, an Na(+)-channel blocker, tetrodotoxin, showed a tendency to enhance ItL-BHGA. On the other hand, ItL-BHGA was enhanced in K+o-free state. 4. Ouabain markedly inhibited ItL-BHGA in both noncompetitive and uncompetitive manners. Benzamil, an inhibitor of the Na(+)-Ca2+ exchange applied simultaneously with ouabain could not prevent ouabain inhibition on ItL-BHGA. The currents induced by other putative neurotransmitters, including a K(+)-dependent Iout caused by dopamine on v-LCDN, were not affected by ouabain. 5. According to our previous study, the threo-L-BHGA receptors are not linked with protein kinases or calmodulin. Then, ItL-BHGA could be produced by the receptor K+ channel complex or the receptor-G-protein-K+ channel combination. The present results indicate that the ATPase activity inhibited by ouabain and the presence of extracellular Na+ and Cl- are needed for threo-L-BHGA to activate the K(+)-dependent structure. Furthermore, the K+o-free state, which inactivates the Na(+)-K+ pump, and tetrodotoxin, which suppresses the Na+ channel at least partly, did not affect the structure to be activated. PMID- 9352314 TI - Effects of cAMP-phosphodiesterase isozyme inhibitor on cytokine production by lipopolysaccharide-stimulated human peripheral blood mononuclear cells. AB - 1. The effects of cAMP-phosphodiesterase (PDE) isozyme inhibitors on the production of tumor necrosis factor alpha (TNF-alpha), and interleukins 1 beta 8 (IL-1 beta and IL-8) by lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMC) were evaluated. In addition, we investigated the effects of dibutyryl cAMP (dbcAMP) and beta-adrenergic receptor agonist on the production of these cytokines. 2. Type IV PDE inhibitors were more effective at inhibiting the production of TNF-alpha and IL-1 beta by LPS-stimulated PBMC than a nonselective, type III or type III/IV inhibitor. In contrast, these agents had no effect on IL-8 production. 3. Increasing concentrations of dbcAMP progressively reduced the production of TNF-alpha and IL-1 beta but not IL-8. 4. The addition of beta-agonist increased the inhibitory effect of PDE inhibitors tested on the production of TNF-alpha and IL-1 beta. 5. Type IV PDE inhibitors could be potent pharmacological agents for the treatment of diseases in which TNF alpha and IL-1 beta are important etiological factors. PMID- 9352315 TI - Effects of amiodarone on ouabain binding to microsomal Na+, K(+)-ATPase in guinea pig heart preparations. AB - 1. The influence of amiodarone on [3H]ouabain (OUA) binding to myocardial Na+, K(+)-ATPase was studied at various KCl concentrations in guinea pig heart microsomal preparations to test the hypothesis that the drug acts on the same receptor sites as cardiac glycosides. 2. First, a series of assays for OUA binding to Na+, K(+)-ATPase were performed in the range of 64-800 nM at 2.5, 5.0 and 10.0 mM K+ concentration. The drug exhibited increasing binding tendency with increasing concentrations and elevation in K+ levels. 3. Competitive binding assays were then performed at 256, 512 and 800 nM OUA in the presence of 50, 100 and 200 microM amiodarone at 5.0 mM KCl, respectively. At each OUA concentration, a concentration-dependent left-to-right shift was observed in the binding affinity with increasing amiodarone concentration. similar assays at 2.5 and 10.0 mM K+ showed the same trends. These effects were significant for 200 mM amiodarone at all K+ levels and OUA concentrations. 4. Furthermore, different OUA concentrations were also shown to displace amiodarone in a concentration dependent fashion. 5. These results indicate that amiodarone competes with OUA for specific binding sites on myocardial microsomal Na+, K(+)-ATPase. They lead to the conclusion that myocardial Na+, K(+)-ATPase is a possible receptor for some of the cardiac actions of amiodarone, such as its proarrhythmic effects. PMID- 9352316 TI - Selective depression of the spinal polysynaptic reflex by the NMDA receptor antagonists in an isolated spinal cord in vitro. AB - 1. The effects of N-methyl-D-aspartate (NMDA) receptor glycine-binding site antagonists 7-chlorokynurenate (7-Clkyn) and (+/-)-3-amino-1-hydroxy-2 pyrrolidone (HA-966) on spinal reflexes in an isolated spinal cord that was maintained in Mg(2+)-free medium in vitro were examined. The actions of 7-Clkyn and HA-966 were compared with those of the channel-site antagonist (i.e., dizocilpine) and NMDA-binding site antagonists--that is, 3-[(+/-)-2 carboxypiperazin-4-yl]-propyl-1-phosphonate (CPP) and DL-2-amino-5 phosphonovalerate (APV). 2. 7-Clkyn and HA-966 produced a selective depression of the polysynaptic reflex (PSR) while negligibly affecting the activity of the monosynaptic reflex (MSR). The PSR was also differentially suppressed by dizocilpine, CPP and APV. The PSR inhibitory activity of the NMDA antagonists was in the following order: dizocilpine > CPP > APV = 7-Clkyn > HA-966. 3. The inhibitory effects of 7-Clkyn on PSR were markedly antagonized by the simultaneous application of D-serine, an agonist for the NMDA receptor glycine binding sites. However, PSR inhibition by dizocilpine and CPP was unaffected. 4. Inhibition of the PSR by 7-Clkyn persisted in the presence of strychnine, which markedly increased the PSR activity by itself. 5. These findings suggest that the NMDA receptor glycine-binding sites play a role in generating the NMDA receptor mediated PSR in the spinal cord in vitro. PMID- 9352317 TI - Effects of S-adenosyl-L-methionine on blood platelet activation. AB - 1. In rats treated with S-adenosyl-L-methionine (SAMe), platelet aggregation in whole blood was inhibited by 50-57% with respect to control values. 2. In whole blood, SAMe also inhibited platelet aggregation, particularly when aggregation was induced with collagen (maximum inhibition 78.6 +/- 5.8% with 10(-5) M SAMe). 3. The antiaggregant response was seen with SAMe in the range of concentrations from 10(-7) to 10(-5) M, whereas concentrations in the range from 10(-4) to 10( 3) M had a progressively weaker effect. 4. Both red blood cells and leukocytes enhanced the antiaggregant effect of SAMe, as did simultaneous incubation with L arginine. 5. The intraplatelet concentration of glutathione also was increased by SAMe. PMID- 9352318 TI - Effects of 4-methyl-2-aminopyridine on [3H]-noradrenaline overflow and contractility of isolated rabbit arteries. AB - 1. The effects of 4-methyl-2-aminopyridine (4M2AP) and 4-aminopyridine (4AP) on spontaneous and evoked [3H]-noradrenaline overflow were compared in rabbit ear artery strips. The effects of 4M2AP on smooth muscle contractility were also investigated in isolated perfused ear arteries. 2. Both 4M2AP and 4AP enhanced spontaneous [3H] overflow from arterial strips in a concentration-dependent manner (10-1000 microM). A bell-shaped dose-response relation was obtained for evoked [3H] overflow over the same concentration range, with maximum effects occurring at 10 microM for 4M2AP (163 +/- 31% increase) and 100 microM for 4AP (154 +/- 16% increase). 3. 4M2AP did not significantly affect evoked tension in the 1-to 100-microM range but clearly depressed it at 1,000 microM (by 65 +/- 11%). In contrast, 4AP enhanced evoked tension in the 10- to 100-microM range (by 30-50%). 4. 4M2AP (10-100 microM) enhanced vasoconstrictor responses to exogenous noradrenaline injections in isolated perfused rabbit ear arteries, whereas higher concentrations (1,000 microM) caused significant depression. 5. 4M2AP (1,000 microM) markedly potentiated vasoconstrictor responses induced by perfusion with a high extracellular K+ solution. When 4M2AP was present during the reloading of noradrenaline-sensitive Ca2+ stores, it enhanced the subsequent vasoconstrictor responses to noradrenaline obtained in a Ca(2+)-free medium. 6. The results show that 4M2AP, like 4AP, enhances [3H] overflow from sympathetic nerve terminals and has complex effects on vascular smooth muscle contractility, indicating the ability of these compounds to affect the Ca2+ permeability of both extracellular and intracellular membrane systems. PMID- 9352319 TI - Adjuvant-carrageenan-induced inflammation in mice. AB - 1. A study was conducted to evaluate the optimum conditions for the induction of adjuvant-carrageenan-induced inflammation (ACII) in Swiss and DBA/1 mice. 2. ACII was induced in mice under experimental conditions similar to those known to be effective in rats. Mice were immunized by subdermal injection of Freund's complete adjuvant (CFA), followed by a subplantar inoculation of carrageenan at different times. 3. The diversities of the responses on ACII between both strains of mice and rats were observed. Data obtained indicate that DBA/1 mice showed an increase in hindpaw and ankle joint swelling, which was more evident on day 21 after carrageenan injection, independently of the time of application of this phlogistic agent. At this time, the histopathological changes were similar to those seen in rats, and were characterized by epidermal hyperplasia, with leukocyte infiltration and granuloma formation. 4. We found that DBA/1 mice, instead of rats, can be used for the evaluation of anti-inflammatory drug activity. However, it is advisable also to consult the histological data to establish whether the synovial changes revert. PMID- 9352320 TI - Suppression of type II collagen-induced arthritis by N-acetyl-L-cysteine in mice. AB - 1. The antiarthritic and anti-inflammatory efficacy of N-acetyl-L-cysteine (NAC) was tested in male DBA/1 hybrid mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index and the phagocytic responses recorded by chemiluminescence in unseparated blood were used for the assessment of disease activity. 2. Mice were immunized by subdermal injection of bovine type II collagen in Freund's complete adjuvant. The treatment with NAC started at day 42 after immunization and was continued over a period of six weeks: in doses ranging up to 50 mg/kg, a dose-dependent suppression of arthritis was noted; between 50 and 200 mg/kg, the inhibition curve had a plateau [ED50 = 50 mg/(kg x day)]. 3. The arthritis index correlated positively with the generation of chemiluminescence by reactive oxygen species (ROS) produced in neutrophils and monocytes activated by 12-O-tetradecanoylphorbol 13-acetate. 4. After treatment with 100 mg/kg of NAC from day 42 after immunization over a period of six weeks, the ROS production was reduced to levels occurring in whole blood of healthy animals. 5. It is concluded that low-molecular-weight antioxidants such as NAC may be adequate for controlling oxidative stress-derived damage in rheumatic diseases by modulation of ROS-dependent signal transduction pathways. PMID- 9352321 TI - Alteration of rat submandibulary gland secretion of protein, calcium and N-acetyl beta-D-glucosaminidase activity by lead. AB - 1. Effects of various doses of long-term lead treatment (0.01%, 0.04% and 0.05%) on rat submandibular saliva were investigated in this study. 2. Both submandibular ducts were cannulated intraorally with polyethylene tubes and saliva was collected from anesthetized lead treated and control rats using pilocarpine as secretagogue. 3. Saliva protein concentration was found to be reduced in lead (0.04%)- and (0.05%)-treated groups. 4. Saliva calcium concentration had a significant reduction only in the lead (0.05%)-treated group. 5. The secretion of the lysosomal enzyme, N-acetyl-beta-D-glucosaminidase (NAG) in saliva decreased significantly in the lead (0.04%)- and (0.05)-treated groups. Specific activity of the enzyme showed an increase in these treated groups. 6. Good correlations were found between saliva protein concentration and NAG activity, saliva protein and calcium concentrations and calcium concentration and NAG activity. 7. There was a correlation between blood and submandibular saliva lead levels, and the saliva/blood ratio was approximately constant for all treated groups. 8. With respect to the ability of lead to substitute for calcium in several intracellular regulatory events, explanation for these alterations in submandibular saliva composition can be made. PMID- 9352322 TI - In vitro administration of 17 beta-estradiol inhibits drug-induced contractions of the rat isolated seminal vesicle. AB - 1. The rat isolated seminal vesicle responded to noradrenaline (NA), acetylcholine (ACh), potassium chloride (KCl) and barium chloride (BaCl2) with reproducible contractions. 2. 17 beta-estradiol (17 beta E) cumulatively added in the isolated organ bath reduced the number of contractions with all agonists used in the rank order of potency: BaCl2 > or = KCl > ACh > NA. The dose-response curves constructed in the presence of 17 beta E (2 x 10(-5) mol/l) produced a rightward shift and a reduction in the maximum response showing inhibitory activity. 3. When the calcium content in the normal Krebs medium (2.5 mmol/l) was reduced to half, the inhibitory activity of 17 beta E was potentiated. The maximum inhibition rates to KCl (phasic and tonic), BaCl2 and ACh were significantly (P < 0.05) different from each other. 4. The inhibitory effects of 17 beta E against all agonists tested were found to be similar in their responses to verapamil, but were much lower in potency. 5. The inhibitory effects of 17 beta E was seen only when the hormone was present in the tissue environment and was readily reversible as soon as the tissue was washed with the Krebs medium, suggesting that the effect of 17 beta E is localized. 6. It is suggested that the in vitro application of 17 beta E on the rat isolated seminal vesicle interferes with the process of translocation of calcium ions from the extracellular medium. PMID- 9352323 TI - Gluconeogenic activity in response to chronic administration of amphetamine sulphate and drug withdrawal. AB - 1. Chronic amphetamine administration (10 mg/kg) to young rats (Rattus norvegius) (40-90 g) produced an initial hyperglycemia following the initial drug administration. This was followed by progressive hypoglycemia with the continuous drug treatment. 2. The present data also demonstrate increases in levels of serum ACTH and corticosterone that were maintained after drug withdrawal, in the case of corticosterone. 3. The Analysis of the transaminase activity revealed the occurrence of significant increases in serum GOT level. Furthermore, progressive increase in the G-6-Pase activity was recorded, reaching its maximum level by the end of the experiment, which demonstrates that induced amphetamine toxicity is time-dependent. PMID- 9352324 TI - Mitoxantrone in the treatment of acute myelogenous leukemia: a review. AB - Mitoxantrone is an intravenous anthracenedione structurally related to the anthracycline antibiotics. This drug has been used for several years in the treatment of acute myelogenous leukemia (AML). Its use has been based on its pharmacological properties, its incomplete cross-resistance with other intercalating agents, and its better tolerance as predicted by preclinical studies. Various treatment schedules, using mitoxantrone alone and in combination with other antileukemic agents, have been used in clinical trials. Complete remission (CR) rates ranged from 14 to 44% in refractory AML and from 46 to 79% in relapsed patients. Although a superiority of mitoxantrone over anthracyclines has not been clearly demonstrated in newly diagnosed patients, mitoxantrone is now recognized as a useful drug in first line therapy. The tolerability profile of mitoxantrone indicates that it offers patients an acceptable quality of life compared with standard treatment regimens, and could be a good alternative to the anthracyclines. The development of new therapeutic concepts aiming at an optimization of its use is now in process and first results are promising. PMID- 9352325 TI - Recent advances in paroxysmal nocturnal hemoglobinuria. From the biology to the clinic. AB - PNH is now known as an acquired, clonal disorder of the hematopoetic stem cells caused by somatic mutation in the X-linked PIG-A gene encoding a protein involved in the synthesis of the glycosylphosphatidylinositol (GPI) anchor by which many proteins are attached to the membrane. Since the past few years, significant advances in the knowledge of the biology of this rare disease have been done. Similarly on the clinical ground, large series of patients with PNH have been published recently, providing estimates of factors affecting survival and of long term follow-up of significant numbers of patients. In this overview we focus on recent advances in the biology and the clinical aspects of this disease, and more importantly try to underline the numerous aspects of yet un-answered questions. PMID- 9352326 TI - The emergence of cell therapy in France. Public health, regulations and other controversial issues. AB - Cell therapy can be defined as the in vivo use of autologous, allogeneic or xenogeneic cells for the prevention, treatment or attenuation of disease. The definition of cell therapy products has been the source of a recent controversy in France. Presently, cell therapy is associated with biological products and manufacturing processes which are frequently poorly defined. A stringent evaluation of cell therapy, as well as the establishment of an optimal regulatory environment, are therefore justified. In France, a recent law (05.28.96) defines cell therapy products as biological products with therapeutic purpose. Cell therapy centers will receive an agreement from the Health Ministry. The French Drug Agency will have responsibility for clinical protocol approval and monitoring, as well as ensure overall quality control. Hopefully, these decisions will contribute to the emergence of cell therapy as a well established, high quality, therapeutic procedure. PMID- 9352327 TI - Cytokine mediated expansion of human umbilical cord blood CD34+ cells: comparison of the use of partially purified and pure CD34+ target cells. AB - To determine the optimal cell population for cytokine mediated expansion, we compared the use of Magnetic Cell Sorting (MACS) system enriched CD34+ human umbilical cord blood (HUCB) cells with that of MACS enriched, flow purified CD34+ HUCB cells. Both MACS enriched CD34+ cells and MACS enriched, flow purified CD34+ cells (mean starting purity of CD34+ SC 51.27 +/- 7.6% and 96.36 +/- 1.34% respectively n = 6) were incubated for seven days with Interleukin-1 (IL-1) + IL 3 + Stem Cell Factor (SCF) and showed a fold increase in the number of nucleated cells (10.02 +/- 2.6 and 18.23 +/- 4.73 respectively) and a reduction in the percentage of CD34+ cells (5.55 +/- 1.23% and 12.21 +/- 3.29% respectively). An increase in the absolute numbers of CD34+ cells (4.8 x 10(4) +/- 2.3 x 10(4)) was observed with MACS enriched CD34+ cells as compared to no change (1.3 x 10(5) +/- 8.8 x 10(4) with MACS enriched, flow purified CD34+ cells. An increase in IL-3 + GM-CSF + SCF responsive colony forming unit (CFU) (1.7 x 10(4) +/- 9.4 x 10(3) and 1.6 x 10(5) +/- 7.7 x 10(4) respectively) was also observed as compared with input values (1.5 x 10(4) +/- 1 x 10(4) and 2.3 x 10(4) +/- 8.9 x 10(3) respectively). We conclude that MACS enriched, flow sorted CD34+ HUCB cells have greater cytokine mediated expansion potential as measured by progenitor expansion, than MACS enriched CD34+ HUCB cells. PMID- 9352328 TI - Parvovirus [correction of Parovirus] B19-induced red cell aplasia in solid-organ transplant recipients. Two case reports and review of the literature. AB - Two solid-organ transplant recipients (one heart and one lung) developed severe anemia with reticulocytopenia. Both were heavily immunosuppressed. Bone marrow aspiration revealed almost complete absence of erythroid precursors. A few giant megaloblastic proerythroblasts with cytoplasmic vacuolisation and intranuclear inclusions were seen. Human parvovirus B19 (B19V)-DNA genome was found by nested PCR assays in blood and bone marrow samples in both cases. Twelve similar cases are described in the literature. When looked for, B19V DNA was positive either in serum or bone marrow or both. Twelve of the fourteen patients were successfully treated by high dose i.v. immunoglobulin (IVIG). One patient recovered spontaneously and another after treatment with recombinant human erythropoietin (rHu-EPO) only. Transplant patients should be considered at risk for severe erythroblastopenic anemia due to B19V infection. Diagnosis is based on bone marrow examination and detection of B19V DNA by PCR in serum and/or marrow. IVIG is an effective and safe treatment. The role of erythropoietin in this indication needs further study. PMID- 9352329 TI - A case of veno-occlusive disease complicated by severe hemorrhagic cystitis successfully treated with recombinant plasminogen activator. AB - Recombinant tissue plasminogen activator (rt-PA) is an effective treatment for veno-occlusive disease (VOD) after bone marrow transplantation (BMT). However rt PA therapy is limited by the risk of hemorrhagic complications. There is little guidance about the use of rt-PA for patients with severe VOD and severe hemorrhage. We report the case of a 16-year-old woman who developed severe VOD associated with life-threatening hemorrhagic cystitis (HC). A dramatic improvement in VOD was obtained after administration of recombinant tissue plasminogen activator (rt-PA). HC was managed with continuous bladder irrigation and blood transfusions. Administration of rt-PA was followed by a moderate increase in blood transfusion requirement but rt-PA did not cause dramatic aggravation of the HC. We conclude that severe HC might not be a contraindication to rt-PA therapy and such patients can be included in randomized trials conducted to determine the efficacy and risk benefit of rt-PA therapy for VOD. PMID- 9352330 TI - Autoimmune thrombocytopenia after six cycles of fludarabine phosphate in a patient with chronic lymphocytic leukemia. AB - Fludarabine phosphate (FDR) has demonstrated a remarkable clinical activity in chronic lymphocytic leukemia (CLL). Myelosuppression is the main toxicity although autoimmune hemolytic anemia (AIHA) is frequently reported. The pathogenesis of AIHA is still unknown however the role of T-cell immunosuppression is suspected. One case of thrombopenia after FDR has been described in a patient with a previous history of an autoimmune thrombocytopenia. We here report a 73-year-old man with a B-CLL and no previous autoimmune disorder who received six courses of fludarabine phosphate and developed afterwards an autoimmune thrombocytopenia. PMID- 9352331 TI - The present state of occupational medicine in Poland. AB - Occupational medicine in Poland has a long tradition, dating back to the establishment of the first health-care institutions for industrial workers at an early stage of Poland's industrialization. Legal foundations of the industrial health-care system, based on the Soviet model, were enacted in 1953. During its most dynamic period of development (1970s and 1980s) the industrial health-care system provided medical services to about 6 million workers. The process of the political and economic transition in Poland began in 1989, and since 1991 there have been numerous transformations in the structure and function of industrial health services. The present article describes the main stages of the transformation process, occupational health training, research, advisory bodies, and the system for setting of hygienic standards. PMID- 9352332 TI - Exposure assessment in occupational epidemiology: measuring present exposures with an example of a study of occupational asthma. AB - The aim of the present paper is to present a comprehensive review of the issues involved in exposure assessment for occupational epidemiology studies and to provide an example. Exposure assessment for occupational epidemiology studies is becoming more quantitatively refined. This paper discusses important issues that need to be taken into account for exposure assessment, with particular reference to occupational asthma. It discusses issues such as survey design, data collection, the effect of measurement error and data interpretation. It presents recently developed methodology to evaluate exposure variability and its effect on the attenuation of risk estimates. It also presents methodology to control for such variability. It uses examples from a recent cohort study of flour millers and bakers. This example shows various characteristics of exposure and demonstrates that various measures of exposure, such as peak and full-shift exposure measurements, are regularly correlated, which has consequences for the analyses of exposure-response relationships. This paper stresses the importance of the recognition and evaluation of exposure variability and its effect on risk estimates and shows that with different exposure grouping schemes, different health risk estimates can be obtained. Quantitative exposure assessment is generally difficult, time-consuming and expensive and many issues need to be taken into account, but it can be rewarding and has become an absolute necessity for many occupational epidemiology studies. Evaluation of components of exposure variance is absolutely necessary. Exposure variability could lead to serious attenuation of risk estimates. PMID- 9352333 TI - Skin absorption of the industrial catalyst dimethylethylamine in vitro in guinea pig and human skin, and of gaseous dimethylethylamine in human volunteers. AB - OBJECTIVES: The aims of the study were three-fold: to assess the skin uptake of the industrial catalyst dimethylethylamine (DMEA) (a) in vitro from water solutions by fresh guinea-pig and human skin specimens, (b) in gaseous form in vivo in human volunteers, and (c) to estimate the relevance of the uptake as an occupational hazard. METHODS: Specimens from the in vitro and in vivo experiments were analysed by gas chromatography using a nitrogen-sensitive detector. DESIGN: DMEA, diluted with water or isotonic saline solution was applied to fresh human or guinea-pig skin, mounted in Teflon flow-through cells with a perfusion fluid flow rate of 1.5 ml/h, samples being collected at 2-h intervals for 48 h. Three healthy male volunteers each had their right forearm exposed (in a Plexiglass chamber) for 4 h to DMEA at each of three different levels (250, 500 and 1000 mg/m3 air). Urine was collected up to 24 h after the start of each experiment. RESULTS: DMEA penetrated both guinea-pig and human skin. The median steady-state flux and permeability coefficient (Kp) values, were 0.009 mg/cm2 x h and 0.001 cm/h, respectively, for guinea-pig skin, and 0.017 mg/cm2 x h and 0.003 cm/h, respectively, for human skin. The median uptake in the three volunteers at the different DMEA exposure levels (250, 500 or 1000 mg/m3) was 44, 64 and 88 micrograms, respectively. The median Kp for all experiments was 0.037 cm/h. CONCLUSION: Uptake of DMEA through the skin is of far less importance than simultaneous uptake via the airways. Thus, the amount of DMEA excreted in urine is a variable of limited use for the purposes of biological monitoring. Although a wide range of Kp values was obtained in the in vitro experiments, both for guinea-pig and human skin, there was no marked difference in median Kp values between the two types of skin. The Kp values were lower than those obtained for human forearm skin in vivo. However, future studies of other tertiary aliphatic amines may show the in vitro method to yield values predictive of those obtained in in vivo studies. PMID- 9352334 TI - Respiratory impairment among children living in the vicinity of a fertilizer plant. AB - The study included 162 second-grade children (85 boys and 77 girls) aged 8-9 years, attending two schools in an area with a fertilizer production plant, and 59 second-graders of the same age (32 boys and 27 girls) from a small neighbouring town located 20 km west of the plant, without any particular source of pollution. During the period from December 1990 to May 1991 the incidence of acute respiratory diseases was surveyed in children and their family members, and forced expiratory volumes were measured in selected second-graders in December 1990 and April 1991. In the area with the fertilizer plant as well as in the compared area ammonia, hydrogen fluoride, nitrogen dioxide, total suspended particulate matter and smoke were measured daily in ambient air and inside the school buildings. The mean concentrations of pollutants during the study period were below the recommended limits, with only a few exceptions, but daily fluctuations, particularly of ammonia and hydrogen fluoride in the area around the plant happened to exceed these values. The observed differences in the levels of air pollution correlated to some extent with the health parameters followed up during the study period. The incidence of acute respiratory diseases corresponded to the registered differences in the exposure to measured pollutants. Forced expiratory volume values in the compared groups of children did not consistently reflect the differences in exposure levels. PMID- 9352335 TI - Determination of carcinogenic potential of mineral fibers by 8 hydroxydeoxyguanosine as a marker of oxidative DNA damage in mammalian cells. AB - 8-Hydroxydeoxyguanosine (8-OH-dG) is a typical form of oxidative DNA damage, which causes mutations in vitro and in vivo. To develop a simple method of testing the carcinogenicity of fibrous materials, the formation of 8-OH-dG was determined in the DNA of J774 cells, an established reticulum cell sarcoma line, after treatment with various natural and man-made mineral fibers. The amount of 8 OH-dG was determined using high-pressure liquid chromatography (HPLC) equipped with an electrochemical detector (ECD). We tested three natural mineral fibers (crocidolite, amosite, and chrysotile) and three man-made mineral fibers (ceramic, glass, and potassium octatitanate). Among them, a significant increase in 8-OH-dG formation was observed in the crocidolite- and amosite-treated cells. We also measured the amount of tumor necrosis factor (TNF) produced by J774 cells incubated with the fibrous materials. Cellular TNF production increased after treatment with all the fibers tested, but it was not statistically significant except in the case of chrysotile. Therefore, these results indicate that the mechanism of TNF production is different from that of 8-OH-dG formation, and that the carcinogenicity of various fibrous materials can be better evaluated by measuring the 8-OH-dG level in J774 cellular DNA after treatment with these fibers. PMID- 9352336 TI - Bronchial hyperresponsiveness and exposure in pig farmers. AB - OBJECTIVE: To study the effect of exposure on bronchial responsiveness in pig farmers. METHOD: A group of 196 pig farmers were tested for lung function and bronchial responsiveness to histamine in the summer of 1992. To achieve sufficient contrast in respiratory morbidity and exposure, 96 of the farmers were selected because they had chronic respiratory symptoms and the remaining 100 because they were free from any respiratory symptoms. Personal exposure to dust, endotoxins and ammonia was measured during 1 working day in the summer of 1991 and 1 day in the winter of 1992. Data on farm characteristics were gathered in the same period. RESULTS: After adjusting for age and smoking behaviour, mild bronchial responsiveness, defined as PC10 < or = 16 mg/ml, was associated with the use of quaternary ammonium compounds as disinfectant [prevalence odds ratio (POR) 6.7, 95% confidence interval (CI) 1.4-32.8], use of wood-shavings as bedding (POR 13.3, CI 1.3-136.7), use of automated dry feeding (POR 2.8, CI 1.0 7.8), use of pellets as feeding material (POR 4.8, CI 1.1-21.1) and location of air exhaust via pit or roof in the confinement units (POR 2.7, CI 1.2-6.3). The association with the use of disinfectants other than quaternary ammonium compounds was not significant (POR 2.4, CI 0.7-8.4). No associations between bronchial responsiveness and measured exposure to dust, endotoxins or ammonia were discernible. CONCLUSION: Protective measures, designed to prevent airway disease in confinement farming, should be based on information about the operational and other characteristics of farms that are related to high exposure and health effects. Specifically, the use of quaternary ammonium compounds as disinfectant, the use of wood-shavings as bedding and the use of automated dry feeding should be discouraged. PMID- 9352337 TI - Urinary excretion of phenols as an indicator of occupational exposure in the coke plant industry. AB - OBJECTIVE: In the present study the relationship between the level of exposure to o-cresol and of 2,4- +2,5-, 3,4-, and 3,5-xylenols and the urinary excretion of their metabolites was examined. The mixed exposure to phenolic derivatives of exposed workers during their work shift was monitored by personal air sampling of the breathing-zone air and by measurements of phenol, o-cresol, and xylenol isomer concentrations in shift-end urine. METHODS: The study subjects were 76 men working at a coke plant who were 22-58 years old and 34 nonexposed subjects. Concentrations of phenolic compounds were determined in the breathing-zone air during the work shift, whereas concentrations of phenol, cresol, and xylenol isomers were measured in urine collected after the work shift. Concentrations of phenols in air and urine were determined by gas chromatography with flame ionization detection. Urine samples were extracted after acid hydrolysis of glucuronides and sulfates by solid-phase extraction. The gas chromatography-mass spectrometry method was applied to identify metabolites in urine samples. RESULTS: The time-weighted average concentrations of phenol, cresol, and xylenol isomers detected in breathing-zone air showed that the exposure level of the workers was relatively low. The geometric mean values were as follows: 0.26 mg/m3 for phenol, 0.09 mg/m3 for o-cresol, 0.13 mg/m3 for p- and m-cresol, and 0.02 0.04 mg/m3 for xylenols at the tar-distillation process. Corresponding urinary concentrations were 10.39, 0.53, and 0.25-0.88 mg/g creatinine for phenol, o cresol, and xylenol isomers, respectively. The correlation coefficients between the o-cresol and 2,4-, 2,5-, 3,4-, and 3,5-xylenol concentrations measured in urine and in the breathing-zone air were statistically significant, varying in the range of 0.54-0.74 for xylenol isomers and being 0.69 for o-cresol. CONCLUSION: We have found that the presence of o-cresol and xylenol isomers in urine can be used as a biomarker for phenol exposure. Analysis performed on workers at the tar-distillation process showed that they were exposed to relatively low concentrations of phenolic compounds. PMID- 9352338 TI - Time-resolved cutaneous absorption and permeation rates of methanol in human volunteers. AB - This paper reports on an experimental study of dermal exposure to neat methanol in human volunteers for the purposes of estimating percutaneous absorption rates, permeation kinetics, baseline (pre-exposure) levels of methanol in blood, and inter- and intrasubject variability. A total of 12 volunteers (seven men and five women) were exposed to methanol via one hand for durations of 0 to 16 min in a total of 65 sessions, making this the largest controlled study of percutaneous absorption for this common solvent. In each session, 14 blood samples were collected sequentially and analyzed for methanol. These data were used to derive absorption rates and delivery kinetics using a two compartment model that accounts for elimination and pre-exposure levels. The pre-exposure methanol concentration in blood was 1.7 +/- 0.9 mg 1(-1), and subjects had statistically different mean concentrations. The maximum methanol concentration in blood was reached 1.9 +/- 1.0 h after exposure. Delivery rates from skin into blood lagged exposure by 0.5 h, and methanol continued to enter the systemic circulation for 4 h following exposure. While in vitro studies have reported comparable lag times, the prolonged permeation or epidermal reservoir effect for such miscible solvents has not been previously measured. The mean derived absorption rate, 8.1 +/- 3.7 mg cm-2 h-1, is compatible with that found in the other in vivo study of methanol absorption. Both in vivo absorption rate estimates considerably exceed in vitro estimates. The maximum concentration of methanol in blood following an exposure to one hand lasting approximately 20 min is comparable to that reached following inhalational exposures at a methanol concentration of 200 ppm, the threshold limit value-time weighted average (TLV-TWA). While variability in blood concentrations and absorption rates approached a factor of two, differences between individuals were not statistically significant. The derived absorption and permeation rates provide information regarding kinetics and absorbed dose that can help to interpret biological monitoring data and confirm mathematical models of chemical permeation. PMID- 9352339 TI - Musculoskeletal complaints in The Netherlands in relation to age, gender and physically demanding work. AB - OBJECTIVES: This cross-sectional study was performed in order to elucidate the relationship of musculoskeletal complaints with age, gender and physically demanding work in the Netherlands. METHODS: Questionnaire data of male (n = 36756) and female (n = 7730) employees, gathered as part of periodical occupational health surveys among active workers in the Netherlands, were stratified for age, gender, and type of work demands. For each stratified group prevalence rates (PR) were calculated for complaints of the back, neck, upper and lower extremities. Moreover, prevalence rate differences (PRD) were estimated as an absolute effect measure of exposure to various types of physical work demands, with active employees in mentally demanding work acting as a reference population. RESULTS: Musculoskeletal complaints among workers in physically demanding occupations were found to increase with age for both sexes. For several complaints, substantially higher rates were reported for women than for men, with a relatively high number of complaints observed among the older female workers (around 40% for complaints of back, upper and lower extremities). Significant PRDs were present in particular for employees in heavy physically demanding occupations and in jobs with mixed mental and physical work demands. CONCLUSIONS: With the ageing of the workforce in mind, these findings stress the need for implementation of preventive measures. Special attention towards the susceptible group of female employees, the elderly age groups in particular, seems justified. In order to clarify the combined effects of age and physical work demands on musculoskeletal complaints, additional studies are required. PMID- 9352340 TI - Risperidone: an analysis of the first three years in general use. AB - Since the introduction in 1993 of the novel serotonin-dopamine antagonist antipsychotic risperidone, over 12 million patient-months of exposure to the drug have been accumulated. Further studies have confirmed the efficacy of risperidone across a broad range of patients with schizophrenia who were not represented in the two pivotal clinical trials. Two studies confirm the efficacy of risperidone in first-episode schizophrenia and subanalyses of these studies suggest that the dose in these patients should be lower than in patients with chronic schizophrenia. In addition, a prospective comparison with risperidone and clozapine and a subanalysis of the North American Trial of risperidone show that risperidone is effective in treatment-resistant schizophrenia. The efficacy of risperidone against negative symptoms has been confirmed by a meta-analysis of seven clinical trials comparing risperidone with active control medication, and by further analyses of the North American Trial (analysis of covariance and path analysis). A long-term open study of risperidone has shown that the benefits of the drug extend well beyond the 8 weeks of the double-blind trials, and the low liability of risperidone for extrapyramidal side effects suggests that patients will be more likely to be compliant with risperidone treatment than with conventional neuroleptic treatment. Relapse rates can therefore be expected to be lower. Evidence from over 1100 patients, 503 of whom had taken risperidone for at least 1 year, suggests that the annual incidence of tardive dyskinesia in patients taking risperidone (7.6-9.4 mg/day) is 0.3%, compared to an annual incidence in patients taking conventional neuroleptics of 5-10%. Studies also suggest that risperidone reduces the number of days that patients with chronic schizophrenia spend as inpatients. It is concluded that mental health-care workers will need to raise their expectations about the treatment outcome of schizophrenia as a result of the introduction of risperidone. PMID- 9352341 TI - Clinical experience in developing treatment regimens with the novel antipsychotic risperidone. AB - Early intervention with antipsychotic treatment has been shown to reduce the risk of relapse and to improve long-term morbidity in patients with schizophrenia. However, treatment with conventional neuroleptics carries with it a significant risk of developing extrapyramidal side effects. Conventional neuroleptics exert their antipsychotic effects at doses similar to those that cause extrapyramidal side effects. Although anticholinergic medication (e.g. benztropine) may reduce the severity of these side effects, anticholinergics are themselves associated with significant side effects, such as dry mouth, constipation, blurred vision, urinary retention and sexual dysfunction. Anticholinergic drugs are also associated with impaired cognition and worsening of the patient's psychosis. Newer antipsychotic drugs, such as risperidone, which have a dual mechanism of action (serotonin-dopamine antagonism) are reported to have a lower risk of extrapyramidal side effects than conventional agents. In particular, risperidone produces significant antipsychotic effects at doses lower than those that cause extrapyramidal side effects. Patients presenting with a first episode of psychosis and who are antipsychotic drug-naive may exhibit abnormal movements before treatment is initiated. Unless these patients are carefully assessed at baseline these movements could be mistaken for drug-induced extrapyramidal side effects. It is important to develop treatment strategies for these patients that improve positive and negative psychotic symptoms as quickly as possible, with a minimal risk of extrapyramidal side effects developing. Prompt and appropriate antipsychotic treatment can substantially reduce the risk of relapse. Whereas approximately 60% of unmedicated patients will relapse in the first year following resolution of the acute psychotic episode, prophylactic antipsychotic medication can reduce this rate to less than 20%. Recent clinical experience in Canada has shown that risperidone is effective in the treatment of patients with first-episode psychosis. Moreover, lower doses of risperidone were required than those previously used to treat chronically ill patients with a history of multiple psychotic episodes and prolonged exposure to conventional neuroleptics. As further data are accumulated, the mean daily dose of risperidone required by first-episode patients has been shown to be close to 4 mg. The low incidence of extrapyramidal side effects in these patients (< 10% required anticholinergic medication) supports the use of risperidone and offers the prospect of improved compliance and a better long-term outcome. PMID- 9352342 TI - Aiding resocialization of the chronic psychotic patient. AB - One difficulty of moving the focus of care for the mentally ill from hospital to the community is that some patients with severe mental illness, who have been in hospital for a long time, are difficult to place in the community. The main problems that appear to be barriers to community-based care are aggressiveness and adverse sexual behaviour. New antipsychotics, such as risperidone, are likely to increase the proportion of patients who can be successfully placed in the community because the low liability for extrapyramidal symptoms will contribute to better compliance. A crucial aspect of the move to the community is the attitudes of neighbours. Studies show that the public is deterred from making contact with the mentally ill because they expect them to have inadequate communication skills. The public also expect bizarre behaviour and aggressiveness. However, the majority of those living near proposed new community homes would welcome the establishment of mental health facilities in their neighbourhood. A study of a public education campaign focussed on the neighbours of a sheltered home found that fear of the mentally ill was reduced and willingness to socialize with them was increased. This change in attitudes was reflected in increased social contact between patients and their neighbours. Therefore, attempts to open community homes for the mentally ill as unobtrusively as possible are likely to lead to more objections than if well constructed public education campaigns focussed on the neighbours are undertaken. PMID- 9352343 TI - Managing the behavioral and psychological signs and symptoms of dementia. AB - As the world's population ages, increasing numbers of patients with dementia can be expected, the signs and symptoms of which can be extremely disruptive. In particular, behavioral and psychological signs and symptoms of dementia reduce the quality of life of carers (usually family members) and increase the cost of care. Conventional neuroleptics have been used for many years in the management of disturbed and disruptive demented patients, although there are few well controlled clinical trials demonstrating their efficacy. The use of the low potency neuroleptics is associated with orthostatic hypotension, cardiac toxicity, anticholinergic side effects and daytime sedation. The high-potency neuroleptics tend to cause extrapyramidal side effects and akathisia. Clozapine although less likely to cause extrapyramidal symptoms than conventional neuroleptics, can cause orthostatic hypotension and requires continual blood monitoring. Early-phase open trials suggest that risperidone is efficacious in patients with behavioral and psychological signs and symptoms of dementia and that it has a low side-effects profile. Further trials are needed to confirm this, but it is likely that the newer antipsychotics, as typified by risperidone, will lead to safer and more effective management of patients with the disruptive and costly behavioral and psychological signs and symptoms of dementia. Non pharmacologic interventions may also provide benefit, though controls are rare. PMID- 9352344 TI - Cognitive function in schizophrenia. AB - Impaired cognitive function in schizophrenia, once thought to be a secondary effect of the psychosis, is now seen as an enduring and core feature. It has many manifestations, but the most disruptive element is arguably a fundamental defect in the patient's ability to manipulate available information. The magnitude of the cognitive deficit in schizophrenia is considerable and remains relatively stable despite fluctuations in other symptoms. The degree of dysfunction also has a high predictive value for long-term disability. In recent years, more attention has been directed towards cognitive dysfunction in schizophrenia as a result of which assessment scales and diagnostic systems increasingly incorporate cognitive dysfunction as an independent domain. Good cognitive function depends upon the brain's ability to prioritize tasks and to switch from parallel processing to sequential processing when the processing load is excessive. This requires working executive memory. Neuroimaging and functional analyses suggest that such cognitive function relies upon unimpaired prefrontal activity. In addition, there is increasing evidence that antipsychotic drugs with 5-hydroxytryptamine (5-HT)2A blocking activity produce better cognitive function in patients with schizophrenia than drugs with predominantly dopamine (D)2-blocking activity (conventional neuroleptics). The development of sophisticated, computer-delivered maze tasks has shown that newer antipsychotics, such as clozapine and risperidone, differ from conventional neuroleptics in their effects on cognitive function. The prospects, therefore, are that patients treated with drugs having 5 HT2A-blocking activity will have better cognitive function and will be better able to function in life's roles than will patients treated with conventional neuroleptics. PMID- 9352346 TI - The generation of encephalitogenic T cell lines from experimental allergic encephalomyelitis-resistant strains of mice. AB - While only a few strains of mice are susceptible to the primary induction or passive transfer of experimental allergic encephalomyelitis (EAE), the basis of EAE resistance remains unclear. In the present studies, we have defined two approaches that allow for the generation of encephalitogenic, myelin basic protein-reactive, T cell lines from EAE-resistant strains of mice. The first approach, based on the putative relevance of apoptosis to autoimmune disease, involves repeat antigenic stimulation of recently initiated T cell lines. The second approach involves the initiation of lymph node cultures in the absence of exogenous splenocytes as antigenic-presenting cells and the use of a higher antigen concentration. Both approaches lead to the generation of encephalitogenic T cell lines from EAE-resistant mouse strains and will be useful for identifying factors relevant to the pathogenesis of EAE. PMID- 9352345 TI - Critical stages of tumor growth regulation in transgenic mice harboring a hepatocellular carcinoma revealed by distinct patterns of tumor necrosis factor alpha and transforming growth factor-beta mRNA production. AB - There is now good evidence that cytokines contribute to the regulation of tumor growth. The cytokine-driven modulation of tumor growth was investigated during the progression of a hepatocellular carcinoma (HCC) in SV40 large T tumor antigen transgenic mice. In vivo, an increased rate of liver growth correlated with increased transforming growth factor (TGF)-beta 1 mRNA expression, while the greatest amounts of tumor necrosis factor (TNF)-alpha mRNA were detected earlier during tumor development. Conversely, no particular alteration of IL-1 alpha, IL 1 beta, IL-6, IL-2, IL-4 and IFN-gamma mRNA production could be reported. In vitro, hepatocyte-like tumor cell lines established at two stages, either before or after HCC differentiation, were characterized. The early-stage-derived cell line produced TNF-alpha mRNA, but had barely detectable expression of TGF-beta 1 mRNA, while later-stage-derived cell lines showed the reciprocal pattern. All cell lines displayed a lack of sensitivity to TNF-alpha, although some degree of sensitivity to TNF-alpha could be observed in the presence of actinomycin-D or after treatment with IFN-gamma. The early-stage-derived cell line was sensitive to the growth inhibitory effects of TGF-beta 1, but late-stage-derived tumor cell lines displayed a loss of sensitivity to TGF-beta 1 which correlated with the increased expression of TGF-beta 1 mRNA. Altogether, this suggests that tumor cells contribute to the discrete TNF-alpha and TGF-beta 1 expression patterns during HCC progression. This model of HCC could be of valuable interest to assess the impact of various immunotherapeutic strategies on modulation of tumor growth. PMID- 9352347 TI - Altered effector responses of H-Y transgenic CD8+ cells. AB - The primary role of CD8+ T cells is to destroy virus-infected or tumor cells expressing cognate antigens in the form of peptide-MHC class I complexes. This destruction is primarily achieved by the actions of lytic mediators and/or lymphokines. In this report, we show that mature, H-Y/Db-specific CD8+ T cells from H-Y TCR transgenic mice were unable to efficiently release lytic mediators after antigenic stimulation. However, anti-TCR antibody induced granule exocytosis and target cell lysis, arguing against signaling and/or cytolytic machinery defects in CD8+ cells, and demonstrating that male antigen induced differentiation of 'naive' into effector CD8+ cells. Stimulation of H-Y-specific effector CD8+ T cells with male stimulators, although insufficient to induce lytic granule release, was sufficient for H-Y-specific IFN-gamma production. Unexpectedly, this effector-phase IFN-gamma production was dependent on B7-2 engagement. We hypothesize that altered effector functions in H-Y-specific CD8+ cells are due to the low affinity of TCR-antigen-MHC interaction and/or the elevated threshold of CD8+ T cell activation. PMID- 9352348 TI - Subcellular localization and translocation of protein kinase C isoforms zeta and epsilon in human peripheral blood lymphocytes. AB - The calcium-independent members of the protein kinase C (PKC) family may play a significant role in T cell function. We have characterized the subcellular localization and redistribution of calcium-independent kinase C activity and of two specific members of this family (zeta and epsilon) in response to activation of human peripheral blood lymphocytes with phorbol myristate acetate (PMA) or through the TCR-CD3 complex. Both PMA and OKT3, an antibody against the TCR associated CD3 complex, induce an increase in membrane and cytoskeletal activity with a concomitant decrease in cytosolic activity. By Western blot analysis, PKC epsilon is present in resting cytosol and membrane fractions, and is detected in the membrane following activation with PMA and in both the membrane and cytoskeleton following OKT3 activation. By contrast, PKC zeta is progressively lost from the cytoskeleton following activation with anti-CD3. Immunocytochemistry reveals distinct redistribution patterns for these enzymes in response to activation through anti-CD3 and by PMA. These findings demonstrate that signaling through the CD3 complex induces significant changes in calcium independent PKC activity and in the intracellular distribution of specific isoenzymes, and support a role for specific functions for individual isoenzymes in T cell activation. Lastly, changes in the cytoskeletal distribution of these isoenzymes suggest a potential role in the modulation of cell structure in response to activation. PMID- 9352349 TI - Control of TCR V alpha-mediated positive repertoire selection and alloreactivity by differential J alpha usage and CDR3 alpha composition. AB - In rats expressing the f allele of the rat MHC (RT1f), CD8 T cells utilizing the V alpha 8.2 segment are 10-fold overselected during thymic development, resulting in V alpha 8.2 expression by 14% of mature CD8 T cells as compared to 1-2% in MHC congenic strains. In the alloreactive responses of CD8 T cells from RT1f-negative rats against RT1f, V alpha 8.2+ CD8 T cells are also preferentially expanded. Neither overselection nor alloreactivity of V alpha 8.2+ TCR require selective V beta pairing. However, RT1f alloreactive V alpha 8.2+ TCR preferentially use a related set of J alpha segments which contribute short homogeneous CDR3 alpha loops, with features suggesting peptide promiscuity, and little N additions. In contrast, only few overselected V alpha 8.2+ CD8 T cells showed an imprint of positive selection on J usage or CDR3 composition. The results demonstrate that a single V alpha segment can promote both MHC allele-specific positive selection and alloreactivity, and that the latter is more dependent on an additional contribution of CDR3 alpha, possibly by promoting reactivity with a diverse set of MHC-bound peptides or by providing additional MHC contacts. PMID- 9352350 TI - Exposure of resting peripheral blood T cells to HIV-1 particles generates CD25+ killer cells in a small subset, leading to induction of apoptosis in bystander cells. AB - Apoptosis is a major mechanism whereby HIV-1 depletes uninfected CD4+ and CD8+ T cells. We previously showed that resting peripheral blood T cells derived from healthy donors were killed by an apoptotic mechanism after adsorption to gp120 containing, protease-defective HIV-1 (L-2) particles, more effectively than parental wild-type LAI adsorption or rgp 120-mediated CD4 cross-linking, followed by mitogenic stimulation. Here, we present evidence that the L-2 particle-based apoptosis was induced both in CD4+ and CD8+ cells by generation of effector cells which were mainly derived from a resting memory CD4+CD38- subset. This subset enhanced the CD25 expression on the surface and secreted IFN-gamma in the culture supernatant after L-2 particle exposure. Significant elevation of Fas ligand mRNA was found in the subset by L-2 particle exposure, while expression of Fas antigen on uninfected T cells was induced by exposure to IFN-gamma. These results indicate that L-2 particles can shift the CD4+CD38- subpopulation from a resting to an activated state, and this activation leads to killing of bystander CD4+ and CD8+ T cells by a Fas-mediated mechanism. In fact, purified CD4+CD38- cells exposed to L-2 particles were converted into effector cells that were able to kill autologous as well as allogenic target T cells pretreated with IFN-gamma. Further, we found that the observation of apoptosis due to L-2 particles was a more general phenomenon, that also occurred with Thai primary HIV-1 isolates. These results suggest that such specific types of HIV-1 particles may play a major role in the induction of apoptosis for both bystander CD4+ and CD8+ T cells, through inappropriate activation of CD4+CD38- cells. PMID- 9352351 TI - Mouse germinal center B cells with the xid mutation retain responsiveness to antimouse CD40 antibodies but diminish IL-5 responsiveness. AB - The germinal center (GC) develops in secondary lymphoid tissues in response to thymus-dependent (TD) antigens. To investigate the molecular mechanism of B cell differentiation in GC, we enriched GC B cells from spleen of TD antigen-immunized wild-type and X-linked immunodeficient (XID) mice, and examined the differentiation of GC B cells into antigen-specific IgG1 antibody-forming cells (AFC) in response to anti-CD40 mAb and cytokines. A significant proportion of freshly purified GC B cells expressed receptors for IL-4 and IL-5. Anti-CD40 mAb sustained the viability of GC B cells and IL-4 co-operated with anti-CD40 mAb for further enhancement of the cell viability. Anti-CD40 mAb and IL-4 were essential for inducing differentiation of GC B cells into antigen-specific IgG1-AFC and IL 5 efficiently enhanced their differentiation. GC B cells with the xid mutation responded for proliferation to CD40 ligation to a lesser extent and for the IgG1 AFC response to anti-CD40 mAb together with IL-4, but they showed impaired responsiveness to IL-5, regardless of enhanced expression of IL-5R in response to anti-CD40 mAb and IL-4. These results suggest that anti-CD40 mAb, IL-4 and IL-5 play a critical role in the differentiation of mouse GC B cells. The GC B cells from XID mice show a functional defect with respect to IL-5-mediated differentiation. PMID- 9352353 TI - Continued differentiation during B lymphopoiesis requires signals in addition to cell survival. AB - During B lymphopoiesis, cells undergo successive rounds of division and growth arrest coupled to intermittent selection on the basis of Ig expression. It is unresolved whether differentiation requires specific signaling or is merely the consequence of sustained cell survival. Transgenic expression of the cell death antagonist, Bcl-2, promoted accumulation of B lymphoid cells in mice deficient in antigen receptor rearrangement (scid or rag-1-/-) and in mice lacking the IgM transmembrane domain (microMT). Continued differentiation occurred, however, only in the bcl-2/scid and bcl-2/microMT mice. The appearance of B lineage cells expressing CD21, CD22 and CD23 was associated with DHJH rearrangements which encode a truncated C mu-containing protein called D mu in bcl-2/scid mice and with expression of Ig heavy chain classes other than IgM in the bcl-2/ microMT mice. In neither case, however, were proliferating cells observed in the more mature B lineage compartments in the bone marrow. Thus, continued B cell development requires signaling via Ig heavy chain-containing receptors and is not simply a consequence of blocking apoptosis. PMID- 9352354 TI - Polymorphism at beta 85 and not beta 86 of HLA-DR1 is predominantly responsible for restricting the nature of the anchor side chain: implication for concerted effects of class II MHC polymorphism. AB - The first hydrophobic pocket, P1, of class II MHC has been shown to be an important site of peptide anchoring. Two polymorphisms occur in this pocket in the human class II MHC beta chain at position 85 and 86. beta 85 is usually Val, occasionally Ala, whereas beta 86 can be Gly or Val. However, Ala85 is found only in conjunction with Val86. The independent effect of the polymorphism at these two positions on the binding of normal and substituted antigenic peptides has never been examined. To do so, three soluble HLA-DR1 variants that contain the naturally occurring combinations of these side chains at these two positions were generated and tested with a panel of influenza matrix peptides varying at anchor P1. DR1 alleles differing only at position 86 are very similar in the binding of a panel of antigenic peptide, indicating that beta 86 does not substantially influence the peptide binding of DR1. In contrast, DR1 varying only at position beta 85 differ in their binding of substituted peptides containing Ala, Tyr or Trp at the P1 anchor position. Thus, beta 85 shows the predominant effect on the P1 anchor side chain preference of the P1 pocket in DR1. This is in contrast to other HLA-DR alleles where beta 86 has been shown to control the nature of the P1 anchor. These previous data together with our own imply that the role of polymorphism in P1 may be influenced by the contextual framework of the remaining allelic polymorphism. PMID- 9352352 TI - Pre-TCR signaling components trigger transcriptional activation of a rearranged TCR alpha gene locus and silencing of the pre-TCR alpha locus: implications for intrathymic differentiation. AB - A rearranged TCR alpha transgene remains transcriptionally inactive in rag-2-/- thymocytes but can be induced by CD3-mediated signals with concomitant maturation of double-negative (DN) thymocytes to the CD4+CD8+ double-positive (DP) stage. Reciprocally, the same signals silence pre-TCR alpha (pT alpha) expression. In normal C57BL/6 thymocytes, TCR alpha expression is not detected in DN thymocytes while, in contrast, TCR beta expression is initiated at the most immature c kit+CD44+CD25- stage and continues throughout thymocyte development. pT alpha expression is first detected at the intermediate c-kit +/- CD44+CD25+ DN stage, increases during transition to the more mature c-kit-CD44-CD25+ stage and is lost at the DP stage. Thus, although TCR beta and pT alpha expression are independent, the pre-TCR complex mediates signals controlling the appearance of alpha beta TCR through selective regulation of TCR alpha and pT alpha genes. PMID- 9352355 TI - Human Ig heavy chain CDR3 regions in adult bone marrow pre-B cells display an adult phenotype of diversity: evidence for structural selection of DH amino acid sequences. AB - Ig repertoires generated at various developmental stages differ markedly in diversity. It is well documented that Ig H chain genes in human fetal liver are limited with regard to N-regional diversity and use of diversity elements. It is unclear whether these characteristics persist in pre-B cell H chain genes of adult bone marrow. Using Ig H chain CDR3 fingerprinting and sequence analysis, we analyzed the diversity of Ig H chain third complementarity determining regions (HCDR3) in adult bone marrow pre-B and mature B lymphocytes. Pre-B cell HCDR3 sequences exhibited adult characteristics with respect to HCDR3 size, distribution of N regions and usage of diversity elements. This suggested that pre-B cells in adults are distinct from fetal B cell precursors with regard to Ig H chain diversification mechanisms. At the DNA sequence level, HCDR3 diversity in mature B cells was similar to that in pre-B cells. Pre-B HCDR3s, however, frequently contained a consecutive stretch of hydrophobic amino acids, which were rare in mature B cells. We propose that highly hydrophobic pre-B HCDR3s may be negatively selected on the basis of structural limitations imposed by the antigen binding site. At the same time, usage of hydrophilic HCDR3 sequences (thought to support HCDR3 loop formation) may be promoted by positive selection. PMID- 9352356 TI - Heparin disaccharides inhibit tumor necrosis factor-alpha production by macrophages and arrest immune inflammation in rodents. AB - Inflammation is the clinical expression of chemical mediators such as the pro inflammatory cytokine tumor necrosis factor (TNF-)-alpha produced by macrophages and other cells activated in the immune response. Hence, agents that can inhibit TNF-alpha may be useful in treating arthritis and other diseases resulting from uncontrolled inflammation. We now report that the cleavage of heparin by the enzyme heparinase I generates sulfated disaccharide (DS) molecules that can inhibit the production of TNF-alpha. Administration of nanogram amounts of the sulfated DS molecules to experimental animals inhibited delayed-type hypersensitivity to a skin sensitizer and arrested the joint swelling of immunologically induced adjuvant arthritis. Notably, the sulfated DS molecules showed a bell-shaped dose-response curve in vitro and in vivo: decreased effects were seen using amounts of the DS molecules higher than optimal. Thus, molecular regulators of inflammation can be released from the natural molecule heparin by the action of an enzyme. PMID- 9352357 TI - The role of I-Ag7 beta chain in peptide binding and antigen recognition by T cells. AB - We examine here how the beta chain of the class II MHC molecule I-Ag7 influences T cell recognition. Three sets of T cell clones were identified. The first set recognizes peptides bound to I-Ag7, I-Ad and I-Ag7 mutant in which the allele specific residues His and Ser at position 56 and 57 were changed to the Pro at residue 56 and to non-polymorphic Asp at residue 57. The second set responds to the antigen presented only by I-Ag7 and does not recognize the peptides bound to the other class II molecules. The third set is also specific for I-Ag7 as a result of the poor binding of the peptide to I-Ad and the mutant I-Ag7. These results indicate that positions 56 and 57 of the I-Ag7 class II MHC beta chain play a role in both T cell recognition of the MHC-peptide complex and peptide binding to MHC. These two different functions may be involved in I-Ag7-restricted beta cell antigen recognition by diabetogenic T cell clones. PMID- 9352358 TI - Agonist peptide modulates T cell selection thresholds through qualitative and quantitative shifts in CD8 co-receptor expression. AB - Engagement of the TCR is a pivotal step in thymocyte development, ultimately resulting in the survival (positive selection) or loss (negative selection) of developing T cells. The roles of peptides and stromal cell interactions necessary for these selection events, however, are still poorly understood. To investigate the effects of agonist peptide in positive selection, we used a novel cell suspension model for in vitro thymic positive selection in adults. Target thymocytes from H-2Db-restricted TCR transgenic mice, specific to the lymphocytic choriomeningitis virus (LCMV) peptide bred on a non-selecting MHC background (H 2d or TAP-1-/-), were co-cultured with freshly isolated H-2b thymic stromal cells. In the presence of selecting stroma the nominal agonist LCMV peptide induced apoptosis at high concentrations and at low concentrations enhanced the efficiency of positive selection both in numbers of cells 'rescued' and kinetics of appearance of selected single-positive cells. We further illustrate down modulation of CD8 alpha beta or CD8 beta at high but non-deleting concentrations of agonist peptide. This highlights the ability of the T cell, within the window of positive selection, to modify surface co-receptors both qualitatively and quantitatively in response to increasing avidity TCR-peptide-MHC interactions. The direct consequence of this would be to lower the total signaling events below the threshold for apoptosis induction. Hence if self peptide were not presented in sufficient quantities in the thymus, autoreactive cells may escape deletion and may actually be positively selected. PMID- 9352360 TI - Suppression of MHC class II expression by human class II trans-activator constructs lacking the N-terminal domain. AB - The class II trans-activator (CIITA) is a bi- or multi-functional domain protein which plays a critical role in the expression of MHC class II genes. We report that removal of the N-terminal 151 amino acids, encompassing all of the acidic domain but leaving intact the proline/serine/threonine-rich domain, results in a mutant protein with potent suppressive properties for MHC class II expression. HeLa cells stably or transiently transfected with mutant CIITA constructs showed up to 99% suppression of MHC class II antigen induction by IFN-gamma and marked suppression of HLA-DRA mRNA expression. Transient transfection of a B lymphoma line resulted in up to 89% reduction of constitutive MHC class II expression within 5 days and suppression of HLA-DRA mRNA synthesis. PMID- 9352359 TI - Targeting Epstein-Barr virus nuclear antigen 1 (EBNA1) through the class II pathway restores immune recognition by EBNA1-specific cytotoxic T lymphocytes: evidence for HLA-DM-independent processing. AB - Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is the only viral protein consistently expressed in all malignancies associated with EBV and there is now convincing evidence to suggest that EBNA1 is not recognized by MHC class I restricted cytotoxic T lymphocytes (CTL). The lack of recognition of EBNA1 has been attributed to a cis-acting inhibitory effect of glycine-alanine repetitive (G-Ar) sequences on the endogenous processing of this antigen through the class I pathway. In the present study we have explored the possibility of targeting EBNA1 through an alternative mechanism using the MHC class II pathway. Using purified EBNA1 protein, we demonstrate here that CD4+ CTL can efficiently recognize EBV transformed B cells and Burkitt's lymphoma cells following exogenous sensitization with this antigen, and this immune recognition is not affected by the G-Ar domain within EBNA1. Analysis of the processing mechanism revealed that intracellular loading of class II molecules with an EBNA1 epitope occurs through an HLA-DM-independent pathway. These results highlight a novel mechanism for immune recognition of EBNA1 and also demonstrate that the G-Ar-mediated protection from processing can be overridden if this antigen is presented through the class II pathway. PMID- 9352361 TI - Detection of precursor Th cells in mesenteric lymph nodes after oral immunization with protein antigen and cholera toxin. AB - We have characterized the earliest antigen-specific Th cells in murine mesenteric lymph nodes (MLN), following oral immunization with the hen egg lysozyme (HEL) as antigen and cholera toxin (CT) as adjuvant. We did this by analyzing in vitro proliferation and cytokine production in response to HEL by the MLN T cells. MLN cells taken 5 days after a single oral immunization with HEL and CT provided the earliest source of proliferating HEL-specific T cells. This proliferation was completely inhibited by anti-IL-2, but not inhibited by anti-IL-4 antibody. IL-2 protein was detected in culture supernatants but not IL-4 using ELISA or bioassays. IL-4 mRNA was not found in responding cells using RT-PCR. Some of the day 5 MLN cultures produced IFN-gamma in response to HEL, but isolated T cells from the same MLN did not. Exogenous IL-4 alone did not stimulate day 5 MLN T cells, but IL-4 did synergize with HEL to induce a large proliferative response. The data indicate that the HEL-specific CD4 T cell pool in MLN 5 days after oral immunization is composed of undifferentiated precursor Th cells. These cells have the potential for IL-2 production and IL-4R expression upon re-stimulation in vitro. PMID- 9352362 TI - Monocyte chemotactic protein-1 is a proinflammatory chemokine in rat skin injection sites and chemoattracts basophilic granular cells. AB - Chemokines may control mast cell infiltrates found in many inflammatory diseases. These cells act through at least two main functions: migration and degranulation. Here we show that human recombinant monocyte chemotactic protein (MCP)-1 (10 ng/50 microliters) induces, after 4 h, an inflammatory vascular permeability and cellular extravasation reaction, determined by Evan's blue dye (1% in saline) injected into the tail vein of the rat, when injected intradermally in the rat skin. The blue color accumulating at the sites of injection provides evidence of vascular permeability and cellular extravasation. The colored areas of the skin were then enucleated and immersed in a fixative solution. Slides were prepared with sections of tissue colored with toluldine blue and analyzed under an optical microscope. A significant number of basophilic cells migrated to the injected area where MCP-1 (10 ng/50 microliters) was used compared to the control PBS treatment. Cell recruitment was slightly less than N-formyl-methionine-leucyl phenylalanine (used at 10(-6) M/50 microliters). Electron microscopy studies confirmed the presence of basophilic granular cells where MCP-1 was intradermally injected. After preparation of a histidine decarboxylase (HDC) probe, a Northern blot analysis was determined for HDC mRNA in the enucleated tissue injected with MCP-1 (10 ng/50 microliters). Steady-state levels of HDC mRNA levels were induced after 4 h. These results were confirmed by the higher amount of histamine release, compared to the control PBS, in the enucleated tissue from the MCP-1 injection sites. Our results suggest that MCP-1 could play a significant role in diseases characterized by basophilic cell accumulation and migration to sites of tissue damage. Moreover, we show for the first time that MCP-1 is a pro inflammatory chemokine that induces basophilic cell migration in rat skin injection sites. PMID- 9352363 TI - Analysis of murine CD22 during B cell development: CD22 is expressed on B cell progenitors prior to IgM. AB - CD22 is a B cell-restricted glycoprotein involved in cell adhesion and signaling. Since CD22 is likely to play an important role in interactions between B cells and other cells, and in regulating signaling thresholds, we characterized the expression of murine CD22 during different stages of B cell development. In contrast to previous reports, we show that CD22 is expressed on B cell progenitors prior to expression of IgM. IL-7-responsive B cell precursors from the fetal liver and early B lineage cells (B220+IgM-) from the bone marrow both express a low density of surface CD22. The majority of the earliest B cell progenitors (B220+IgM-CD43+) in the bone marrow, however, do not express CD22. As B cells mature, the density of CD22 molecules on the cell surface increases. B220brightIgM+ bone marrow cells express high levels of CD22, as do splenic B cells. The correlation of CD22 levels with B cell maturation is replicated in an in vitro culture system, which distinguishes stages of B cell development based on function. Following activation of mature resting splenic B cells with anti-mu mAb or lipopolysaccharide (LPS), levels of CD22 decrease. Finally, we show that the addition of anti-CD22 mAb augments the proliferative response of both anti-mu and LPS-stimulated B cells, suggesting a role for CD22 in diverse signaling pathways. PMID- 9352364 TI - Constitutive activation of NF-kappa B in an animal model of aging. AB - The pathophysiology of many disease states observed in aged individuals has been linked to the dysregulated production of several pleiotropic cytokines. We have demonstrated that NF-kappa B, a major transcriptional regulator of these aberrantly expressed cytokines, exists in a constitutively activated state in cells obtained from the major lymphoid organs of aged animals. Therapeutic treatment with dietary antioxidants or with agents capable of activating the peroxisome proliferator-activated receptor (PPAR)-alpha was able to correct the abnormal nuclear NF-kappa B activity, reduce lipid peroxide levels, and eliminate the dysregulated expression of cytokines and other genes under NF-kappa B control. These results suggest that abnormal activation of NF-kappa B in aging contributes to the dysregulated expression of certain pleiotropic cytokines. Effective therapeutic regimens for aging and other inflammatory disease states might benefit from the administration of antioxidants or other agents which specifically activate PPAR-alpha. PMID- 9352365 TI - Role of IL-2 in rat fetal thymocyte development. AB - Early during rat thymus ontogeny, an important proportion of thymocytes expresses IL-2R and contains IL-2 mRNA. To investigate the role of the IL-2-IL-2R complex in rat T cell maturation, we supplied either recombinant rat IL-2 or blocking anti-CD25 mAb to rat fetal thymus organ cultures (FTOC) under several experimental conditions. The IL-2 treatment initially stimulated the growth of thymocytes and, as a result, induced T cell differentiation, but the continuous addition of IL-2 to rat FTOC, as well as the anti-CD25 administration, resulted in cell number decrease and inhibition of thymocyte maturation. These results indicate that immature rat thymocytes bear functional high-affinity IL-2R and that IL-2 promotes T cell differentiation as a consequence of its capacity to stimulate cell proliferation. Modifications in TCR alpha beta repertoire and increased numbers of NKR-P1+ cells, largely NK cells, were also observed in IL-2 treated FTOC. Furthermore, IL-2-responsiveness of different thymocyte subsets changed throughout thymic ontogeny. Immature CD4-CD8-cells responded to IL-2 in two stages, early in thymus development and around birth, in correlation with the maturation of two distinct waves of thymic cell progenitors. Mature CD8+ thymocytes maximally responded to IL-2 around birth, supporting a role for IL-2 in the increased proliferation of mature thymocytes observed in vivo in the perinatal period. Taken together, these findings support a role for IL-2 in rat T cell development. PMID- 9352366 TI - Peptide-induced deletion of CD8 T cells in vivo occurs via apoptosis in situ. AB - The ultimate fate of T cells undergoing antigen-induced cell death in vivo remains controversial. Whereas apoptosis of CD4+ T cells driven by superantigen is readily detectable in lymphoid organs, CD8+ T cells have been reported to disappear from the lymphoid organs and accumulate in the liver where they undergo apoptosis. Using transgenic mice that produce large numbers of ovalbumin-specific CD8+ T cells (OT-I cells), we were able to investigate the events that follow soluble peptide administration in an independent CD8+ T cell system. Here we show that the OT-I cells undergo proliferation and apoptosis in situ in lymphoid organs in response to antigenic stimulation with no evidence for liver involvement. This is similar to the course of events found for CD4+ T cell activation and counters the view that the liver is a general site for CD8+ T cell clearance following antigen-specific activation. PMID- 9352367 TI - Molecular genetic characterization of XRCC4 function. AB - XRCC4 is a generally expressed protein of 334 amino acids that is involved in the repair of DNA double-strand breaks and in V(D)J recombination, but its function is unknown. In this study, we have used a mutational approach and the yeast two hybrid method to perform an initial characterization of this protein. We show that the XRCC4 protein is located in the nucleus. We also demonstrate that several potential phosphorylation sites are not required for XRCC4 function in a transient V(D)J recombination assay. In addition, we show that XRCC4 forms a homodimer in vivo with the homodimerization domain being located within amino acids 115-204. Finally, we define a core domain of XRCC4 that functions in V(D)J recombination and comprises amino acids 18-204. Potential functions of XRCC4 are discussed. PMID- 9352368 TI - Specificity of helix-induction by 2,2,2-trifluoroethanol in polypeptides. AB - The specificity of helix-induction in polypeptides by 2,2,2-trifluoro ethanol (TFE) is studied using an all beta-sheet protein such as cardiotoxin analogue I (CTX I) from the Taiwan Cobra (Naja naja atra) and a homopolymer such as poly-L lysine. It is found that alcohols including TFE can 'non-specifically' induce helix at high concentrations both in CTX I and polylysine at neutral pH. However, among the alcohols used, only TFE could transform the heat-induced beta-sheet conformation of polylysine at pH 11.5 into an alpha-helix. The beta-sheet to alpha-helix conversion in polylysine (in the beta-sheet conformation) occurs even at very low concentrations of TFE (< 5% v/v). In addition, experiments on the effect(s) of TFE on the denatured and reduced CTX I (rCTX I) indicate the helix induction in rCTX I takes place at low TFE concentrations (< 20% v/v). The results of this study hint at the possible influence of disulfide bridges on the induction of helix by TFE. PMID- 9352369 TI - Thermodynamic behaviour of gliadins mixture and the glass-softening transition of its dried state. AB - The glass-softening transition of a mixture of gliadins extracted from wheat flour has been studied in its dry state by differential scanning calorimetry (DSC). Further, the rate of removal of its water vapours on its evaporation from a gliadins mixture containing different amounts of water has been investigated, and through this the presence of any exothermic effect that could be attributed to polymerization of gliadins has been examined. The heat absorbed in this evaporation is comparable with the heat of evaporation of pure water measured in a separate experiment in identical conditions. This showed that the gliadins mixture did not polymerize on heating up to 473 K in the presence of moisture. In this respect the behaviour of the gliadins mixture differs remarkably from that of gluten studied before (J Phys Chem 1996:100:19692). The effects of purge gas, helium and argon, on the calorimetric effects during the evaporation of water have been studied. A restudy of gluten shows that helium decreases substantially the endothermic signal in the DSC measurements, and thereby reveals the exothermic effects of polymerization in gluten, but argon does not do so. The structural relaxation time, t, of dry gliadins mixtures at different temperatures has been calculated from an analysis of its glass-softening endotherm. The temperature at which t = 1 ks is 452 K, and the Tg, obtained by the usual method of intersection of the straight lines drawn, is 443 K, 7 K higher than for the polymerized dry gluten, the distribution of relaxation time parameter is 0.25, and increase in the heat capacity in this range is 0.21 J/g K. Physical ageing effects are considerable in the gliadins mixture, which alters the glass softening endotherm but not the structural relaxation time or its distribution. PMID- 9352370 TI - An evaluation of crystal structure of mannan I by X-ray powder diffraction and molecular mechanics studies. AB - The present study reports the re-examination of the crystal structure of mannan I by the X-ray powder diffraction study combined with the miniature crystal model simulation. A primary objective of this study was to investigate an adequate chain staggering position along the fiber axis. Among the several crystal structure models proposed for mannan I, that derived from the electron diffraction study of the single crystals was adopted as a starting model. The X ray crystallographic residuals were calculated for the single crystal structure at different chain staggering positions, while the ab projection was kept invariant. In a similar fashion, the miniature crystal models consisting of seven mannotetraoses were constructed, each having different chain staggering values and the three-dimensional structures of all the models were subsequently optimized by using the MM3(92) program without introducing any constraint. Both the X-ray residual and lattice energy plots with respect to the chain staggering position showed the common minima around the -0.25 x c chain staggering. The minimum models were subjected to a search for preferred orientations of O2 and O6 hydroxyl groups. It was found that the hydroxyl groups present inside the minicrystal tended to rotate into particular orientations during the structure optimizations to form the O5-H-O2 and O3-H-O6 intermolecular hydrogen bonds between the adjacent oligomers in an alternative direction. PMID- 9352372 TI - Examination of polypeptide beta-sheet structure in solutions and thin layers: determination of the concentration and the 'critical aggregation concentration' using a cyanine dye as sensor. AB - In the visible spectra of some cyanine dyes a bathochromic shift of the dye monomer band was observed on the preconditions that: (1) beta-sheet containing polypeptides (denotes also proteins) were presented; and (2) these polypeptides were embedded in a layer or aggregated in solution. The band with the polypeptides which contained only the alpha-helix did not shift. In several cases the absorbance lifetime of the shifted band was limited. This was caused by dye self-association at the polypeptide surface, but there were enough quantities for this lifetime to obtain exact analytical measurements. These were executed quantitatively (100-20% beta-sheet), qualitatively (to about 10% beta-sheet) and moreover for the determination of the 'critical aggregation concentration' (cac). The applications of the dye sensor in biophysics, medicine and pharmacy were discussed. PMID- 9352371 TI - Scanning tunnelling microscopy observation of cytochrome-c denaturation induced by bromopyrogal red on highly oriented pyrolytic graphite. AB - The denaturation of cytochrome-c (cyt-c) induced by bromopyrogal red (BPR) was studied by scanning tunnelling microscopy (STM) on the electrochemically pretreated highly oriented pyrolytic graphite (HOPG) surface. STM images reveal that denatured cyt-c molecules exist in variable states including aggregates, globular compact, partially unfolded and combined with BPR molecule. The apparently low image contrast of denatured cyt-c observed in this experiment comparing to that of native cyt-c molecules, and the relative low image contrast of the unfolded part comparing with the compact globular part, are ascribed to the unfavourable tunnelling paths for the conformational variations of denatured cyt-c molecules. PMID- 9352373 TI - The muscle motor: 'simultaneous' levers or sequential impulses? AB - We use the step-size distance equation z = u/n developed in our two previous papers (z is the step-size distance, u is the actin filament relative velocity and n is the rate of ATP splitting on a given actin filament), and introduce one additional concept: that the impulsive contractile forces developed on an actin filament should proceed sequentially along a given actin-myosin train. This enables us to elucidate some unexplained and puzzling data in the literature, and to predict the surprisingly high values of ATPase in intact muscle that have recently been found experimentally. It seems that a sequential impulsive model of the actin myosin interaction may give a better explanation of many phenomena in muscle physiology than does the current model of the action of simultaneous levers. PMID- 9352374 TI - Sequence similarity between xylose isomerase and replicase: another TIM-barrel in the replicase structure? AB - The BLAST search using the strand beta 2 (46_GAHGVTFHDDDLIP) of the (alpha/beta)8 barrel of xylose isomerase from Streptomyces olivochromogenes resulted in retrieving the sequentially similar segment of replicase from garlic latent virus (692_GGHGIGFHRDD). The detailed analysis of the entire amino acid sequences of both xylose isomerase and replicase suggested that the polypeptide segment 644 1046 of replicase (the entire length of this enzyme is 1924 residues) could share the structure of xylose isomerase (20.7% identity using the entire sequence of xylose isomerase). The relatedness of replicase and xylose isomerase is supported by the fact that the sequence similarity can be observed along the whole sequence of xylose isomerase (386 amino acid residues). The sequence of replicase exhibits moreover the similarity with that of lycopene cyclase, an enzyme implicated in the beta-carotene biosynthesis, that was previously found to share similarity with xylose isomerase. Thus the relevant segment of replicase is predicted to adopt an (alpha/beta)8-barrel topology similar to that of xylose isomerase. PMID- 9352376 TI - Hemostatic tests in the prediction of atherothrombotic disease. AB - Hemostatic components play major roles in the pathogenesis of human atherothrombotic disease. There has been interest in determining whether tests for hemostatic components predict this disorder. Recent population-based and clinically-oriented prospective studies have begun to provide useful information. Northwick Park Heart (NPH) study made the initial observation that the plasma fibrinogen level is an independent risk factor for non-fatal and fatal coronary heart disease (CHD). This has been confirmed by other population-based studies. By contrast, factor VIIc levels which were reported by NPH to be an independent risk factor for CHD, especially fatal myocardial infarction (MI) has not been confirmed by other studies. Factor VIIIc, von Willebrand factor (vWF), tPA and PAI-1 are reported to be associated with CHD. Prospective angina pectoris studies have identified fibrinogen, C-reactive protein (CRP) and von Willebrand factor as independent risk factors for acute MI. These findings raise a possible mechanism of inflammation in CHD. A number of studies imply an association of platelet activation with CHD. A prospective study has shown that a persistent positive spontaneous platelet aggregation (SPA) test is an independent risk factor for recurrent MI. Hence, prospective studies indicate a potential value for certain hemostatic tests to predict atherothrombotic disorder. However, clinical utility of these tests remains to be established. Controlled clinical trials may be required to provide a more definite information regarding the use of these tests for selecting high risk patients for preventive strategies. PMID- 9352375 TI - Hyperhomocysteinemia: a risk factor for arterial and venous thrombotic disease. AB - Patients with the rare homozygous hereditary defects of homocysteine metabolism that cause severe hyperhomocysteinemia and homocystinuria are at high risk of arterial and venous thrombosis. This prompted studies of the relationship between moderate hyperhomocysteinemia and thrombotic risk in the general population. In the last 2 decades, retrospective case-control studies and prospective cohort studies have demonstrated moderate hyperhomocysteinemia to be a frequent and independent risk factor for premature vascular disease in the coronary, cerebral, and peripheral arteries. More recently, the association of moderate hyperhomocysteinemia with venous thrombosis was shown in patients with early onset or recurrent disease and in the general population. Genetic and environmental factors act in concert to cause moderate hyperhomocysteinemia. Since inadequate intake of folic acid, vitamin B12, or vitamin B6 are most frequently associated with hyperhomocysteinemia, dietary supplementation of these vitamins could have a tremendous impact on the epidemiology and natural history of arterial and venous thrombotic diseases. PMID- 9352377 TI - Prediction of postoperative venous thrombosis using haemostasis tests. AB - The prediction of patients who are at sufficiently high risk of postoperative deep venous thrombosis to indicate perioperative antithrombotic prophylaxis has traditionally used only clinical risk factors. The associations of preoperative and/or postoperative haemostatic tests with postoperative deep venous thrombosis are reviewed. In general, the results support the biological concept of a preoperative and postoperative prothrombotic tendency in patients who develop deep venous thrombosis. Increased levels of coagulation activation markers and decreased assays of fibrinolytic potential show consistent relationships to postoperative deep venous thrombosis. At present, however, the clinical utility of such tests is unproven; so that at present they cannot be advocated for routine preoperative or postoperative screening. PMID- 9352378 TI - Gamma delta T-cells in human cutaneous immunology. AB - Gamma delta T-Cells represent a minor subpopulation of T-lymphocytes in man and their role in normal and diseased human skin is unknown. This article is a comprehensive review of T-lymphocytes bearing the gamma delta T-cell receptor in normal and pathological human skin. Firstly, we have documented the occurrence of gamma delta T-cells in normal skin and in a range of reactive and malignant skin conditions. We have then discussed the experimental findings regarding the repertoire used by gamma delta T-cells in normal human skin and in cutaneous disorders with an increased percentage of gamma delta T-cells. PMID- 9352379 TI - Acquired transplant tolerance. AB - Increasing the acceptance rate of organs is the central goal of transplantation research. Long-term survival of vascularized organs without chronic immunosuppressive therapy has been achieved in experimental animals. In humans, the possibility of achieving immunological tolerance and a drug-free state has been reported occasionally in patients who after withdrawal of immunosuppressants because of major toxicity still carry a functioning graft. It has been proposed that organ transplant implies a migratory flux of donor 'passenger' leukocytes out of the graft into the recipient tissue or organs, to establish a persistent condition of 'microchimerism'. Although there is evidence that the same migratory mechanisms apply to all organ grafts, migration of 'passenger' leukocytes is less in kidney and heart than in liver. To enhance the acceptance of organs less tolerogenic than liver, perioperative infusion of donor bone marrow has been attempted to increase the donor 'passenger' leukocyte load. It has been suggested that the established microchimerism is not only associated with long-term acceptance of the graft, but it also plays an active role in induction and maintenance of donor-specific unresponsiveness. However, the intimate mechanism(s) responsible for prolonged graft survival in this setting remain speculative. Experimental evidence is also available that the thymus plays a major role in the development of self-tolerance and is critical in the induction of acquired tolerance to exogenous antigens. It has been reported that after intrathymic injection of donor cells clonal deletion of maturing thymocytes occurs and is the major mechanism in the induction of donor-specific tolerance, since peripheral T-cell component would be devoid of alloreactive population. Studies are warranted in the near future to explore whether the thymus technique can be employed to prolong survival or induce tolerance to allograft in humans. An interesting novel strategy for transplant tolerance is also the oral administration of alloantigens, which has been recently applied to the cardiac transplant model in rat. All these approaches will have a major impact in the near future on transplant medicine, opening new perspectives to obtain indefinite graft survival. PMID- 9352381 TI - Peripheral neutrophils after allergic asthmatic reactions. AB - The response of peripheral neutrophils was studied in 16 patients with allergic asthma after challenge with birch/grass pollen allergen, in order to identify inflammatory markers associated with only the early asthmatic reaction and those associated with both early and late asthmatic reactions. The allergen challenge proceeded until the patients had an early asthmatic reaction with 100% increase in specific airway resistance. Bronchoconstriction after allergen challenge was monitored hourly over 9 h and finally after 18 h, by measurement of the forced expiratory volume in 1 s. Seven patients had a late reaction, defined as a decrease in forced expiratory volume in 1 s of more than 15%. Blood samples were taken before and 18 h after challenge. After allergen challenge (18 h) the blood concentration of neutrophils in patients with a late asthmatic reaction was 1.4 times higher than before challenge and there was a tendency for increased Fc gamma receptor-mediated chemiluminescence. Lewis X-antigen (CD 15), which is associated with endothelial adhesion and extravasation, significantly decreased at the same time. Neutrophils were incubated with the tetrapeptide arginine glycine-aspartate-serine before and 18 h after allergen challenge. Both patient groups showed an increased Fc gamma receptor-mediated chemiluminescence and a decreased Fc gamma receptor membrane expression following allergen challenge, suggesting a preactivation. In conclusion, patients with a dual asthmatic reaction show a sustained primed inflammatory response and primed neutrophils compared with patients with only an early reaction when measured after the decline of clinical symptoms provoked by allergen challenge. PMID- 9352380 TI - Metabolic and hemodynamic effects of peptide leukotriene C4 and D4 in man. AB - The time course of the effects of intravenous or intracoronary administration of peptide leukotrienes on metabolic parameters and on systemic and coronary hemodynamics was evaluated in 15 patients with normal coronary arteries. Peptide leukotriene C4 (2 nmol given as a bolus intravenous injection) induced an early fall (at 2 min) in mean arterial pressure (P < 0.02) associated with a rise in heart rate (P < 0.001) and in plasma levels of epinephrine (P < 0.05) and norepinephrine (P < 0.005), but without significant changes in coronary blood flow or coronary vascular resistance. Mean arterial pressure, heart rate, norepinephrine, and epinephrine returned to baseline values 10 min after leukotriene C4 administration. In contrast, at 10 min post leukotriene C4, with coronary blood flow and myocardial oxygen consumption unchanged, an increase in coronary vascular resistance (P < 0.05) and in myocardial oxygen extraction (P < 0.01) was observed, which returned to baseline values at 20 min post leukotriene C4. Peptide leukotriene D4 (3 nmol, given in the left coronary artery) induced an early (20 s) and transient fall in mean arterial pressure (P < 0.001) paralleled by a rise in heart rate and plasma levels of epinephrine and norepinephrine, all of which returned to baseline at 10 min. Coronary vascular resistance increased at 10 and 15 min (P < 0.02 and P < 0.05, respectively) and myocardial oxygen extraction at 15 min (P < 0.02). These results suggest that small doses of peptide leukotrienes induce both an early and transient fall in mean arterial pressure associated with secondary sympathoadrenergic activation, and a late increase in small coronary arteriolar resistance. PMID- 9352382 TI - Polymorphonuclear leukocytes from asthmatics release more calcium from intracellular stores and have enhanced calcium increase after stimulation with N formyl-methionyl-leucyl-phenylalanine. AB - Polymorphonuclear leukocytes isolated from peripheral blood of asthmatics appear to be primed to release more reactive oxygen species than cells of healthy subjects. The enhanced agonist-induced rise in the intracellular free calcium concentration may be responsible for this increased respiratory burst. To test this hypothesis we studied the N-formyl-methionyl-leucyl-phenylalanine- and cyclopiazonic acid--(an inhibitor of Ca(2+)-ATPase of intracellular calcium stores) induced calcium increase in the polymorphonuclear leukocytes of 28 subjects (16 with moderate asthma, 69.6% +/- 8.3% predicted normal peak expiratory flow and 12 normal controls) using a fluorescent probe Fura-2AM at 100 nM and 1 mM extracellular calcium concentrations. In 1 mN calcium, the N-formyl methionyl-leucyl-phenylalanine-induced calcium increase was 1.7-fold higher in asthmatics than in healthy subjects. Similarly, the contribution of calcium from intracellular stores to the calcium response to N-formyl-methionyl-leucyl phenylalanine was higher in asthmatics (55% +/- 14% vs. 39% +/- 14%, P < 0.01). The pool of calcium released from intracellular stores by N-formyl-methinoyl leucyl-phenylalanine and cyclopiazonic acid was 2.3- and 2.2-fold larger than in control cells. There was a correlation between maximal intracellular calcium concentration related to N-formyl-methionyl-leucyl-phenylalanine-induced calcium release from intracellular stores and forced expiratory volume in 1 s expressed as percentage predicted and reversibility in asthmatics (r = 0.63, r = -0.53, P < 0.05). In conclusion, polymorphonuclear leukocytes of asthmatics exhibit an altered calcium response that is mainly dependent on increased calcium release from intracellular stores. PMID- 9352383 TI - Bioavailability of Desmin, a low molecular weight dermatan sulfate, after subcutaneous administration to healthy volunteers. AB - The bioavailability of two different s.c. doses of Desmin (a new low molecular weight dermatan sulfate) was evaluated in 12 healthy volunteers (6 men, 6 women aged 22-45 years) who were injected, on 3 separate days and with a wash-out period of at least 21 days between each administration, with 200 and 300 mg of Desmin by the s.c. route and 200 mg by the i.v. route. Immediately before injection and at various times thereafter (after 15 min and 30 min for i.v. only and after 1, 2, 3, 4, 6, 8, 12, and 24 h for both s.c. and i.v. dosing), blood samples were drawn to investigate bioavailability by measuring several coagulation parameters: activated partial thromboplastin time, thrombin time, inhibition of factor Xa, Heptest, and heparin cofactor II. Furthermore the local tolerance of the s.c. and i.v. injections were investigated. The s.c. administration of the two Desmin doses had a negligible effect on the activated partial thromboplastin time and a very small effect on the thrombin time, measured with human thrombin; in contrast, Heptest, heparin cofactor II, and anti Xa activities increased, with a good drug bioavailability (more than 100%). The plasma effects of Desmin were dose dependent only when measured by Heptest, which also gave a greater response after the s.c. administrations. There were no symptoms of intolerance or pain at the injection site after single i.v. and s.c. Desmin administration. PMID- 9352384 TI - Influence of bisphosphonate on the negative erythropoietic effects of uranyl nitrate. AB - Uranium salts, such as uranyl nitrate, induce severe renal dysfunction and tubular necrosis and a significant impairment of both oxygen dependent erythropoietin production and response to recombinant human erythropoietin. All effects are transient and reach maximal severity on the 7th day post injection. We investigated the effects of ethane 1-hydroxy-1, 1-bisphosphonate, which counteracts the inhibitory effect of uranyl nitrate on bone formation, on the negative erythropoietic effects of uranyl nitrate. Adult female Wistar rats received 1 mg/kg body weight of uranyl acetate by the i.v. route. Ethane 1 hydroxy-1,1-bisphosphonate was injected simultaneously at a dose of 7.5 mg/kg by the same route. Seven days after drug injections, plasma erythropoietin was estimated after hypobaric hypoxemia or cobalt chloride administration. The response to exogenous erythropoietin was also measured in uranyl nitrate- and/or ethane 1-hydroxy-1,1-bisphosphonate-injected rats made polycythemic by transfusion. The erythroid response was quantitated in terms of red blood cell 59iron uptake. Ethane 1-hydroxy-1, 1-bisphosphonate counteracted the effect of uranyl nitrate on oxygen-dependent and cobalt-dependent erythropoietin production, but did not correct the right shift of the dose-response relationship for exogenous erythropoietin induced by uranyl nitrate in the polycythemic rat. PMID- 9352385 TI - Distribution of IgG subclasses after anti-hepatitis B virus immunization with a recombinant vaccine. AB - To assess whether a different IgG subclass distribution was elicited in "low" and "high responders" after vaccination with recombinant hepatitis B virus surface antigen, we selected from 360 vaccine recipients 30 "low-responder" subjects, with anti-HBs levels of 10-160 mIU/ml, and 40 "high-responder" subjects, with anti-HBs levels greater than 10,000 mIU/ml. In both groups all IgG subclasses were elicited in the anti-HBs response and the greatest contribution was that of IgG1, followed by IgG2. IgG1 was significantly less represented after the second (58%) and third doses (61%) of vaccine in "low responders" compared with "high responders" (65% and 69%). The relative percentage of IgG2 was significantly higher after the second (33%) and third (30%) doses of vaccine in "low responders" than in "high responders" (29% and 26%). In "low responders" the age of vaccine recipients significantly influenced the anti-HBs IgG subclass distribution: IgG2 and IgG4 production was positively correlated with age, whereas the opposite was observed for IgG1. These data support the evidence that: (1) IgG1 and IgG2 subclasses are mainly involved in the specific anti-HBs response both in "high" and "low responders"; (2) the relative contribution of specific IgG2 to vaccination is higher in low responders and progressively increases with age. PMID- 9352387 TI - Human pharmacology and PK/PD modelling. PMID- 9352386 TI - Liver is not the unique site of synthesis of beta 2-glycoprotein I (apolipoprotein H): evidence for an intestinal localization. AB - Apolipoprotein H is a protein of about 50 kilodaltons, structurally related to the regulators of the complement activation family. Its physiological function is poorly understood but it has been implicated in lipid metabolism and coagulative pathways. The major site of synthesis is thought to be the liver. Several reports indicate that apolipoprotein H is the antigen of the antiphospholipid antibodies and also behaves as an acute-phase reactant. Moreover, 40% of plasma apolipoprotein H is associated with very low-density lipoprotein, high-density lipoprotein, and postprandial chylomicrons. In this study we investigated other sites of synthesis by reverse transcription/polymerase chain reaction and we found apolipoprotein H mRNA expression in intestinal cell lines and tissues. Immunohistochemistry was performed on various fresh and paraffin-embedded tissues and apolipoprotein H was immunolocalized in the cytoplasm of hepatocytes and epithelial cells from colon and jejunum. This study indicates that apolipoprotein H is expressed at both mRNA and protein levels in enterocytes. PMID- 9352388 TI - Basic concepts of pharmacokinetic/pharmacodynamic (PK/PD) modelling. AB - Pharmacokinetic (PK) and pharmacodynamic (PD) information from the scientific basis of modern pharmacotherapy. Pharmacokinetics describes the drug concentration-time courses in body fluids resulting from administration of a certain drug dose, pharmacodynamics the observed effect resulting from a certain drug concentration. The rationale for PK/PD-modelling is to link pharmacokinetics and pharmacodynamics in order to establish and evaluate dose-concentration response relationships and subsequently describe and predict the effect-time courses resulting from a drug dose. Under pharmacokinetic steady-state conditions, concentration-effect relationships can be described by several relatively simple pharmacodynamic models, which comprise the fixed effect model, the linear model, the long-linear model, the Emax-model and the sigmoid Emax model. Under non steady-state conditions, more complex integrated PK/PD-models are necessary to link and account for a possible temporal dissociation between the plasma concentration and the observed effect. Four basic attributes may be used to characterize PK/PD-models: First, the link between measured concentration and the pharmacologic response mechanism that mediates the observed effect, direct vs. indirect link; second, the response mechanism that mediates the observed effect, direct vs. indirect response; third, the information used to establish the link between measured concentration and observed effect, hard vs. soft link; and fourth, the time dependency of the involved pharmacodynamic parameters, time-variant vs. time-invariant. In general, PK/PD-modelling based on the underlying physiological process should be preferred whenever possible. The expanded use of PK/PD-modelling is assumed to be highly beneficial for drug development as well as applied pharmacotherapy and will most likely improve the current state of applied therapeutics. PMID- 9352389 TI - Connection of pharmacokinetics and pharmacodynamics--how does it work? AB - The present article gives an introduction to the correlation between drug transports and drug action in a body system. Direct linked model uses the law of mass action to describe the PK/PD modelling. More complex models like indirect linked models are discussed. PMID- 9352390 TI - PK/PD modelling of high-dose diltiazem--absorption-rate dependency of the hysteresis loop. AB - To investigate bioequivalence of 2 different sustained-release diltiazem formulations the preparations each containing 180 mg diltiazem-HCl were given to 20 healthy male volunteers in an open, randomized, 2-way crossover design. Blood samples were taken before drug administration and at 14 times until 30 hours post application. 12-lead ECGs were recorded at the same time points, and atrioventricular conduction time was monitored as a safety parameter. Plasma samples of 8 subjects were assayed by HPLC. Peak values of plasma concentrations and prolongation of the PQ interval were taken from the plasma concentration or ECG data directly, AUCs of pharmacokinetic and pharmacodynamic effects were calculated by the linear trapezoidal rule, MRTs were calculated as the first moment over AUC. Bioequivalence was tested according to Schuirman (ratios of parameters and shortest 90% confidence intervals) using pharmacokinetic and pharmacodynamic data sets. Relative bioavailability of the test preparation with respect to AUC0-30 was 110% with the 90% confidence interval ranging from 100 to 130%. Bioavailability with respect to Cmax was significantly higher (190%) with a 90% confidence interval not even including 100%. Consequently, MRT was significantly lower with the test preparation (80%), again with a confidence interval not including unity. Relative bioavailability of the test product in terms of pharmacodynamic parameters was 160% in the extent of the effect (AUEC0 10), 190% even with the rate of the effect (Emax) and 80% with the mean residence time (MRTE). All parameters differ significantly between the products, Bioinequivalence was therefore concluded from these results. The functional relationship between pharmacokinetic and pharmacodynamic parameters could be described by hysteresis loops, however, with a clockwise rotation. This cannot be explained in the classical way by the time-lag between central and effect compartments. Two alternative conceivable explanations, namely formation of antagonistic metabolites or downregulation, were tested for plausibility. A comparison of the present results to literature data favors the model of downregulation/tolerance development. This model is additionally supported by the finding that the shape of the hysteresis is dependent on the absorption rate of diltiazem, calculated as mean input time according to MIT = MRT - 1/lambda z. It is concluded that acute tolerance develops at least with the electrophysiological action of diltiazem after oral application and that the extent of tolerance development increases when decreasing its absorption rate. Bioequivalence assessment of diltiazem is possible using pharmacodynamic parameters, however, since PK/PD relationships are influenced by the absorption rate, extent parameters may be misinterpreted when rate parameters of the test formulations are different. PMID- 9352391 TI - Pharmacokinetic-pharmacodynamic modelling of the in vitro antiinfective effect of piperacillin-tazobactam combinations. AB - PURPOSE: The aim of the study was to investigate the in vitro antiinfective effect of piperacillin-tazobactam (PIP-TZB) combinations on Escherichia coli in simulations of free concentration time profiles of both drugs, similar to those obtained in human tissue after i.v. bolus administrations. METHODS: An in vitro dilution model was used to expose E. coli ATCC 35218 (beta-lactamase producer) to various piperacillin-tazobactam concentration profiles obtained after i.v. bolus multiple dose, using different dose ratio combinations (1:4, 1:8, 1:16) and dosing regimens, ranging from once-a-day to 4 times a day. The antimicrobial effect was evaluated by determination of the number of bacteria over time. The concentration of PIP in the model was determined by HPLC. RESULTS: A modified Emax model was used to describe the pharmacodynamic effect. The model was linked with the piperacillin concentrations determined experimentally to provide a pharmacokinetic-pharmacodynamic (PK-PD) model. The EC50 for piperacillin alone averaged 5.66 +/- 0.29 micrograms/ml. The EC50 for all doses of piperacillin combined with 0.5 g of tazobactam were dose-dependent and averaged 1.70 +/- 0.56, 3.95 +/- 1.02, and 6.14 +/- 1.24 micrograms/ml for PIP 2, 4, and 8 g, respectively. By increasing the dose of TZB in combination with a fixed dose of PIP, a decreased EC50 was observed. CONCLUSIONS: The PK-PD model allowed a detailed evaluation of the dosing regimens investigated. The results suggested that for these combinations, 3 times a day administration is as effective as 4 times a day. Pharmacodynamic activity of the combinations can be prolonged by sufficiently high inhibitor concentrations. PMID- 9352393 TI - Analysis of drug-receptor interactions in vivo: a new approach in pharmacokinetic pharmacodynamic modelling. AB - Analysis of pharmacodynamic data using the empirical Hill equation only provides limited insights in the underlying factors that determine the shape and location of a concentration-effect curve, such as agonist affinity and efficacy. We have developed a method which allows for the estimation of agonist affinity and efficacy in vivo and yields more insight in the factors that determine pharmacodynamic variability. The method is based on the "operational model of agonism", which describes agonist concentration-effect curves in terms of the maximum system effect (Em), the slope of the transducer function (n), the agonist dissociation equilibrium constant (KA) and an efficacy parameter (tau). We applied the model to obtain estimates of apparent affinity and efficacy of a series of N6-cyclopentyladenosine (CPA) analogues for adenosine A, receptor mediated in vivo effects on heart rate in rat [Van der Graaf et al. 1997]. In all cases, the model converged and estimates of apparent affinity (pKA) and efficacy (tau) were obtained which were highly consistent with results from in vitro radioligand-binding studies. In conclusion, we have shown that the operational model of agonism can provide meaningful measures of agonist affinity and efficacy in vivo. The model may serve as a practical guide for future development of partial adenosine A1 receptor agonists and help to elucidate the mechanisms underlying adenosine A1 receptor-mediated responses in vivo. PMID- 9352392 TI - Pharmacodynamic and pharmacokinetic properties of an angiotensin II receptor antagonist--characterization by use of Schild regression technique in man. AB - OBJECTIVE: The pharmacodynamic properties of a new angiotensin II receptor antagonist (BAY 10-6734) in humans were to be quantitatively characterized from the rightward shifts of the agonist dose-response curves after administration of different doses of the antagonist. METHODS: 24 healthy male volunteers received single oral doses of 20-300 mg BAY 10-6734. Before and up to 23 h post dosing (p.d.) plasma was obtained for HPLC measurement of parent compound and active metabolite BAY 10-6735. Exogenous angiotensin II was infused in increasing dose steps until blood pressure had increased by +25 mmHg. Angiotensin II dose response curves were fitted individually using the sigmoidal Emax model. From the antagonist-induced rightward shifts, as compared to a premedication curve, dose ratios (DR) were determined and DR-1 plotted versus applied dosages and measured plasma concentrations. From these Schild regression plots the fictive doses and concentration (Ki) inducing a DR-1 = 1, i.e. a 2-fold shift in agonist dose response curves, were derived. The "doubling (t2.0) time" of the apparent Ki doses was calculated. RESULTS: BAY 10-6734 dose-dependently induced rightward shifts of the angiotensin II blood pressure response curves, mean maximum DR at 2 h p.d. ranged from 42 (80 mg) to 216 (300 mg), and at 23 h p.d. decreased to about 2 (80 mg) to 4 (300 mg). Pharmacodynamic (3.4-4.6 h) and pharmacokinetic half-lives (3.4-4.3 h) were nearly identical. Apparent Ki doses increased from about 1-2 mg at 2 h p.d. to about 80-100 mg at 23 h p.d., their time course revealed a doubling (t2.0) time of 3.5-3.8 h. A Ki concentration of about 10 micrograms/l was obtained for the active metabolite BAY 10-6735. CONCLUSIONS: Oral administration of BAY 10-6734 in man antagonized angiotensin II dose blood pressure response curves in a dose-dependent manner. The time kinetics of the pharmacodynamic effect, derived from the decay of DR-1 values, as well as the doubling time of the apparent Ki values well agreed with the pharmacokinetic half life. Schild regression revealed competitive angiotensin II antagonistic properties within the dose/concentration range tested. This technique was shown to be an adequate means to evaluate pharmacodynamic potency and kinetic behavior of an angiotensin II receptor antagonist in vivo. PMID- 9352394 TI - Pharmacokinetics and pharmacodynamics of triamterene and hydrochlorothiazide and their combination in healthy volunteers. AB - Although triamterene has been in clinical use for over 30 years, the linearity of triamterene kinetics was not systematically tested. Moreover, although triamterene is mostly applied concomitantly with thiazide-type diuretics the interaction of triamterene (TA) with hydrochlorothiazide (HCT) is subject to a controversial discussion. Therefore, the aim of this study was to examine the dose linearity of TA and the pharmacokinetic and pharmacodynamic interaction of triamterene and hydrochlorothiazide. In the first study 10 healthy volunteers received 0, 12.5, 25, 50, and 100 mg triamterene orally in a balanced crossover design. In the second study 0, 25, and 50 mg TA with 12.5, and 25 mg HCT, respectively, were administered to 12 healthy volunteers. Urine volume and concentration of sodium, TA, hydroxytriamterene sulfate (OH-TA ester), and HCT were measured by flame photometry and thin-layer chromatography, respectively. The observation period for each treatment was 3 days and the drug was given on the second day. Sodium excretion was increased by both drugs. Renal excretion of both TA and OH-TA ester seemed to be reduced at higher doses. However, statistical evaluation revealed no significant (p = 0.37, and p = 0.20, respectively) deviation from linearity. Renal excretion of HCT was not affected by TA and vice versa. However, renal excretion of OH-TA ester is significantly reduced when HCT is administered concomitantly. The renal excretion rate of sodium can be described by a common Emax model when the effects of the excretion rates of both TA and HCT are additive. It is concluded that the pharmacokinetics of TA is linear within the tested dose range and that pharmacodynamic additivity of HCT and TA is not due to a pharmacokinetic interaction. The results support the hypothesis of a sequential nephron blockade for both drugs acting on different tubular segments. PMID- 9352395 TI - Inverse PK/PD: estimation and differentiation of bioavailability from effect kinetics--observations with beta-adrenoceptor antagonists. AB - The time course of pharmacodynamic effects allow to resolve bioavailability relevant pharmacokinetic information, provided simple assumptions can be made about their interrelation. This approach furthermore allows to differentiate ancillary properties of chemically distinct pharmacological agents on the basis of their mutual pharmacodynamic actions. The advantages and pitfalls of this strategy are discussed here for beta-adrenoceptor antagonists on the basis of their pharmacodynamic characterization by the extent of their binding ability to ex vivo in vitro beta-adrenoceptors (RRA assay) and their blunting effects on the ergometric rise in heart rate. PMID- 9352396 TI - Chronopharmacological aspects of PK/PD modelling. AB - Nearly all physiological functions as well as pathophysiological events display reproducible rhythmic changes within 24 hours of a day, including the cardiovascular system. Clinical chronopharmacological studies with antihypertensive drugs gave evidence that effects on the rhythms in blood pressure and heart rate are also dependent on the time of day. Chronopharmacokinetic studies with propranolol, oxprenolol, nifedipine, verapamil, etc. also revealed daily variations in the drugs' kinetics. In general, Cmax was higher and/or tmax shorter after morning than evening dosing of these rather lipophilic drugs. However, independently of whether or not daily variations in the kinetics were found the dose-response relationship was always dependent on the time of day. These data demonstrate that PK/PD modelling has to take into account reproducible rhythmic variations both in the kinetics and/or effects of cardiovascular active drugs. PMID- 9352397 TI - Complex PK/PD models--an alcoholic experience. AB - The absorption and disposition of ethanol are among the more complex of challenges for pharmacokinetics. Attempts to explain the relative bioavailability of ethanol in beer of differing ethanol concentrations have led to models of first pass extraction which recognize the concentration dependent nature of ethanol elimination as well as the role of hepatic blood and the rate of absorption from the gut. The pharmacodynamics of ethanol have also proven to be of matching complexity. While effects on motor coordination only required a delay between plasma and response time profiles the effects on a cognitive test revealed the rapid development of tolerance in addition to a delay. Description of the time course of ethanol effects in an individual after a single oral dose requires over 10 parameters and offers a serious challenge for forensic predictions. PMID- 9352398 TI - Exploring clinical study design by computer simulation based on pharmacokinetic/pharmacodynamic modelling. AB - Computer simulations have been successfully applied in various industries (e.g. automobile, aerospace) to make product development more efficient. Just recently, it was suggested to use simulations in support of clinical drug development for predicting clinical outcomes of planned trials. The methodological basis for this approach is provided by pharmacokinetic and pharmacodynamic mathematical models together with Monte Carlo techniques. In the present paper, the basic notions of clinical trial simulation are introduced and illustrated with the example of an oral anticancer drug. It is shown that computer simulation helps to evaluate consequences of design features on safety and efficacy assessment of the drug which are not easily obtained otherwise. An overview of existing simulation resources with respect to training and software is provided. PMID- 9352399 TI - PK/PD simulations as a tool for rational design of clinical dosage regimens: an example with Fradafiban. AB - In clinical studies, pharmacodynamic effects should be achieved, e.g. maintenance of certain effects with reversibly acting drugs. Available data base for early phase II studies is frequently kinetics in healthy volunteers from phase I studies and pharmacodynamic effects from in vivo or ex vivo data. Using the fibrinogen receptor antagonist Fradafiban as an example, a procedure to achieve rational dosage regimens is described. Applied methods were: curve fitting of plasma concentrations obtained in phase I studies, correlation of fibrinogen receptor occupancy (FRO) to plasma concentrations using a sigmoid Emax model with Hill coefficient for PK/PD correlation, simulation of time course of FRO for various dosage regimens, using estimates for variability from PK and PD data in order to estimate not only mean values but also expected range of FRO. In a phase II study with Fradafiban administered intravenously, therapeutically active plasma levels had to be achieved rapidly and maintained over 24 hours employing a simple infusion regimen in 20 patients and 3 dose groups. For the target dose, a FRO of 80% should be achieved in most patients. Using the tools mentioned above, a rapid initial infusion rate of 10 mg over 30 minutes, followed by a maintenance infusion of 30 mg for the remaining 23.5 hours for the target dose resulted in a median predicted FRO of 82%. For the lower dose, a 5/15 mg infusion over 0.5/23.5 hours, achieving a predicted FRO of 68% was used and the upper dose (15/45 mg) resulted in 87% FRO. Median experimental results were 84% FRO for the target dose and 73% and 88%, respectively, for the lower and upper dose. As these results fit reasonably well to the predictions, it can be concluded that kinetics in the mostly elderly patients is similar to kinetics in healthy young volunteers. Furthermore, FRO in patients is nearly identical to that in spiked human plasma. Taken together, it could be proven that PK/PD methods are a useful tool for design of clinical studies of Fradafiban. PMID- 9352400 TI - Clinical PK/PD modelling as a tool in drug development of corticosteroids. AB - Corticosteroids are used for the treatment of a variety of different diseases both locally and systemically. Most therapeutic effects result from glucocorticoid receptor-mediated events, and there seems to be no substance specific difference in the post-receptor reaction cascade. Therefore, the extent and duration of glucocorticoid effects depend only on the availability of the respective steroid at the receptor site and its affinity to the receptor. This makes glucocorticoids an ideal candidate for PK/PD modelling. Availability at the receptor site is governed by pharmacokinetic parameters such as bioavailability, clearance, protein binding, and volume of distribution. The receptor affinity can easily be measured in vitro. A suitable indirect-response PK/PD model is presented that allows description of the receptor-mediated drug effects such as endogenous cortisol suppression as a function of time. Furthermore, this model allows prediction of the systemic activity of newly developed corticosteroids based on their pharmacokinetics and their respective receptor-binding affinity. The model can also be applied in order to study systemic steroid effects after topical administration or to investigate the effect of the time of dosing on cortisol suppression. Comparison of predictions based on this model and results from large clinical studies are in excellent agreement. Corticosteroids may represent an ideal class of drugs for the successful use of PK/PD modelling during drug development allowing to save time and expenses. PMID- 9352401 TI - Advances in the cutaneous manifestations of thyroid disease. PMID- 9352402 TI - Women in dermatology--we couldn't do without them. PMID- 9352403 TI - Skin and treponemal diseases among Asian domestic house-helpers in northern Saudi Arabia. AB - BACKGROUND: Asian domestic house-helpers in Saudi Arabia come from a different socioeconomic setting with a different disease pattern from that of their host country. This study reports the incidence of skin and treponemal diseases in this group seen at a referral hospital in northern Saudi Arabia. METHODS: The study was based on the analysis of the dermatologic and serologic examinations of 1520 domestic house-helpers during resident permit issue, and a retrospective study of clinical records of house-helpers with skin disorders. RESULTS: Routine examination revealed significant skin disease in 374 (24.6%) individuals, and the disease was transmissible in 126 (8.3%). Treponemal infection (5, 0.3%) and leprosy (1, 0.07%) were seen. Hand dermatitis and chicken pox were the most common causes of hospital attendance. Psychologic skin disorders included three cases of neurotic excoriations, two cases of delusion of parasitosis, and a case of dermatitis artefacta. CONCLUSIONS: The prevalence of transmissible skin diseases in Asian domestic house-helpers is low compared with that in their home countries. Excluding individuals with stigmata of atopic dermatitis from employment as house-helpers, adequate counselling will reduce the incidence of hand dermatitis and psychologic skin disorders. There is a need for continuous surveillance to prevent the introduction of skin diseases not normally seen in the native population. PMID- 9352404 TI - Skin colonization of Staphylococcus aureus in atopic dermatitis patients seen at the National Skin Centre, Singapore. AB - OBJECTIVE: This prospective study sought to determine the bacterial colonization rates on eczematous and non-eczematous skin and nasal mucosa of patients with atopic dermatitis attending a tertiary dermatologic referral clinic in Singapore. The colonization rates were evaluated according to age, sex, race, and severity of dermatitis compared with controls. The results may help to determine whether antibiotics should be considered in the treatment of atopic dermatitis. PATIENTS: Patients, of any age, presenting with atopic dermatitis at the subsidized clinic of the National Skin Centre, Singapore, between 23 August 1996 and 14 September 1996, were included in the study. RESULTS: Thirty-three patients with atopic dermatitis were seen at the outpatient clinic during the study period. Staphylococcus aureus was isolated in 69.7% of the eczematous lesions and in 42.4% of non-eczematous skin of patients with atopic dermatitis. S. aureus was isolated in 53% of patients with mild dermatitis, and in 100% with moderate and severe dermatitis. The nasal carriage rate of S. aureus was higher in atopic dermatitis patients (51.5%) than in non-atopics (35%) (not significant). S. aureus was isolated in 42% of non-eczematous skin in atopics compared with only 5% in the control group (p = 0.003). In patients with atopic dermatitis, all S. aureus isolated was sensitive to cloxacillin, cephalexin, clindamycin, and co trimoxazole; 92% was sensitive to erythromycin, but only 13% was sensitive to penicillin and ampicillin. In the control group, all S. aureus isolated was sensitive to cloxacillin, cephalexin, erythromycin, clindamycin, and co trimoxazole, but only 13% was sensitive to penicillin and ampicillin, and 87% to tetracycline. CONCLUSIONS: This study confirmed that the skin of patients with atopic dermatitis was more frequently colonized with S. aureus than that of non atopics. The more severe the dermatitis, the higher the rate of colonization. S. aureus is also more of than present in non-eczematous skin of atopics than of non atopics. There is also a higher percentage of S. aureus nasal carriage in patients with atopic dermatitis than in non-atopics. Hence antibiotics may have a role in the treatment of atopic dermatitis. Because 87% of S. aureus is resistant to penicillin and ampicillin, antibiotics such as cloxacillin and cephalexin should be used to eradicate S. aureus in the skin of atopic dermatitis individuals. PMID- 9352405 TI - Two feet-one hand syndrome: a retrospective multicenter survey. AB - BACKGROUND: The two feet-one hand syndrome is not uncommon; however, there have only been a few reports on this condition. This study was undertaken to obtain a better understanding of the epidemiology of the two feet-one hand syndrome. METHODS: A retrospective chart review was conducted of all the patients seen in our practices over the past 15 years with the diagnosis of two feet-one hand syndrome. RESULTS: A total of 80 patients with mycologically confirmed disease were identified (men, 72 (90%); women, 8 (10%); 77 (96%) Caucasian; 3 (4%) African-American; age (mean +/- standard error (SE)), 55.9 +/- 2.1 years). The mean age of the patients when the physician was first seen for the condition was 51.3 +/- 2.0 years. The mean ages when the symptoms first developed on the feet and hand were 37.1 +/- 2.4 years and 45.7 +/- 2.2 years, respectively. Tinea pedis was found to occur at an earlier age than tinea manuum (t(65) = 6.92, P < 0.01). There was a significant relationship between the hand in which tinea manuum developed, the hand used to excoriate the soles of feet (chi 2(4) = 14.82, P < 0.01), and the hand used to pick toenails (chi 2(4) = 14.82, P < 0.01); however, there was no significant relationship between handedness and the development of tinea manuum in the dominant hand. The occupation of the patient at the time of development of the two feet-one hand syndrome was categorized according to whether the intensity of hand use was high, moderate, or low. Patients with a high intensity of hand use in their jobs were significantly more likely to develop tinea pedis/onychomycosis (r = -0.27, F(1,61) = 4.77, P < 0.05) and tinea manuum (r = -0.30, F(1,62) = 6.31, P < 0.05) at an earlier age. The best multiple predictors of the age at which medical attention was sought were the age of onset of tinea manuum and a family history of tinea infection (r = 0.86, F(2,59) = 86.9, P < 0.01). The age of onset of tinea manuum was the best single predictor, with a correlation of 0.85. CONCLUSIONS: In the two feet-one hand syndrome, the development of tinea pedis/onychomycosis generally preceded the development of tinea manuum. Tinea manuum usually developed in the hand used to excoriate the feet or pick toenails. Patients whose occupation involved a high intensity of use of the hands were more likely to develop the disease at an earlier age. Patients were more likely to seek attention once tinea manuum had developed, particularly if there was a family history of tinea infection. PMID- 9352406 TI - Tinea capitis in south-western Ethiopia: a study of risk factors for infection and carriage. AB - BACKGROUND: Tinea capitis is a common dermatophyte infection which constitutes an important public health problem among children worldwide. The endemic nature of scalp ringworm in Africa is perpetuated mainly by the lack of knowledge about the prevalence and carrier status, and the absence of control measures. METHODS: Two hundred and nineteen schoolchildren from urban and rural communities of the Illubabor district, south-western Ethiopia, were examined, and scalp samples were taken. Children were classified according to clinical signs and mycologic findings. RESULTS: Physical examination revealed that 29% of the children had clinical lesions compatible with tinea capitis. Dermatophytes were isolated from 33% of the children's scalp samples; of these, 16% had clinical lesions and 17% were identified as carriers. Trichophyton violaceum was responsible for 97% of infections. CONCLUSIONS: Tinea capitis was the second most prevalent cutaneous finding in these children, with a higher prevalence in the urban community; the predictive value of the clinical diagnosis was low and a high proportion of children were identified as carriers in these communities. No relationship between household overcrowding and scalp infection was found. PMID- 9352407 TI - A retrospective study of the clinical presentation and outcome of herpes zoster in a tertiary dermatology outpatient referral clinic. AB - BACKGROUND: This is a retrospective study of the epidemiology and morbidity of herpes zoster and the risk factors for herpes zoster morbidity in Singapore. RESULTS: The mean age of 164 patients with herpes zoster seen at our dermatology clinic between January 1994 and December 1995 was 48.8 years, with a sex ratio of 1:1. The common presenting symptoms were pain (90%), feelings of helplessness and depression (20%), and flu-like symptoms (12%). The commonest prodromes were pain (41%), itching (27%), and paresthesia (12%). Prodromal pain was more frequently experienced by patients aged more than 50 years (42%) than by patients aged less than 30 years (25%). The thoracic (45%) and cervical (23%) dermatomes were the most commonly affected in all age groups. There was no statistically significant difference in the frequency of dermatomal distribution among the different age groups and between the sexes. Pain was experienced by almost all (95%) patients during the course of their disease. It tended to be more severe in older patients. Burning (26%), stabbing (15%), and shooting (15%) pain were the most common types experienced. Post-herpetic neuralgia was significantly more common in older patients. The prevalence of post-herpetic neuralgia decreased over time in all age groups. A higher proportion of older patients (more than 50 years of age) (20%) suffered from post-herpetic neuralgia compared with younger patients (less than 30 years of age) (7%) (not significant). Patients in all age groups considered acute pain (46%) and persistent pain (25%) to be their most unbearable symptoms during the course of herpes zoster. The most significant problems caused by herpes zoster pain were insomnia (25%), misery (feeling helpless and depressed) (20%), limitation of movement (9%), and inability to continue work (8%). Insomnia was significantly more commonly experienced by patients more than 50 years of age (36%) than those less than 30 years of age (P = 0.026). Few patients (9%) consulted their general practitioner (GP) during the prodrome or on the day of appearance of skin eruptions. Most patients (45%) consulted their GP within the first 3 days of the onset of skin eruptions; 33% sought treatment more than 3 days after the appearance of zoster symptoms. Only 30% of patients were willing to pay more than S$200 for antiviral therapy. Most (43%) were only prepared to pay for antiviral treatment if it cost less than S$200. The most important features the patients wished to derive from antiviral therapy were a shortening of the duration of skin lesions (55%) and a reduction in the severity of pain (acute and chronic) (30%). CONCLUSIONS: Our study indicated that older patients (aged more than 50 years) were at a higher risk of developing post herpetic neuralgia. They were also more likely to suffer morbidity, e.g. insomnia. There is a need to educate patients at risk to identify the prodrome and skin eruptions of herpes zoster so that early antiviral therapy can be considered. PMID- 9352408 TI - The role of human papillomavirus in the development of pyogenic granulomas. AB - BACKGROUND: Pyogenic granulomas (lobular capillary hemangiomas) and condyloma acuminata share similar locations and risk factors. Human papillomavirus (HPV) types 6 and 11 are commonly associated with condyloma acuminata, but their association with pyogenic granulomas has not been evaluated. The purpose of this study was to determine whether pyogenic granulomas contain evidence of infection with condyloma-producing HPVs. METHODS: Polymerase chain reaction assays for the E6 and E7 gene sequences of HPV types 6 and 11 and another assay for the E7 region of HPV types 16, 31, 33, 35, 42, and 58 were used to evaluate deoxyribonucleic acid (DNA) extracted from archival pyogenic granuloma biopsies taken from cutaneous and oral epithelium. RESULTS: Neither cutaneous nor oral pyogenic granulomas contain amplifiable E6 or E7 sequences from any of these viruses. CONCLUSIONS: Pyogenic granulomas are not caused by HPV 6, 11, 16, 31, 33, 35, 42, or 58. This study does not exclude the possibility that other viruses may be responsible for these tumors. PMID- 9352409 TI - Neonatal skin lesions due to a spirochetal infection: a case of congenital Lyme borreliosis? PMID- 9352410 TI - Faint erythema. Another manifestation of cutaneous sarcoidosis? PMID- 9352411 TI - Lymphocutaneous infection due to Scedosporium apiospermum. PMID- 9352412 TI - Atypical acropustulosis in infancy. PMID- 9352413 TI - Febrile ulceronecrotic Mucha-Habermann's disease with fatal outcome. PMID- 9352414 TI - Bilateral scrotal extramammary Paget's disease in a Chinese man. PMID- 9352415 TI - Progressing dermatofibromas following surgery. PMID- 9352416 TI - Albendazole: a new therapeutic regimen in cutaneous larva migrans. AB - BACKGROUND: Various therapeutic modalities have been used to treat cutaneous larva migrans, including physical treatments (cryotherapy), topical drugs (tiabendazole), and systemic drugs (tiabendazole, albendazole, and ivermectin). Physical treatments are often ineffective and not devoid of side-effects. Topical tiabendazole is difficult to find in many countries; it is effective orally but frequently causes side-effects. Ivermectin has been used in a small number of patients. METHODS: Eleven (six men and five women) adult patients with cutaneous larva migrans characterized by multiple and/or diffuse lesions were treated with oral albendazole (400 mg daily for 7 days). No other topical or systemic drugs were used and no physical treatment was given. RESULTS: All patients were cured at the end of treatment. No side-effects were complained of or observed, and no laboratory abnormalities were recorded. No recurrences were observed. CONCLUSIONS: Albendazole is effective in the treatment of cutaneous larva migrans characterized by multiple and/or diffuse lesions. This new therapeutic regimen can reduce the number of no responses and recurrences, sometimes observed following shorter (e.g. 3-5 days) treatments with albendazole. The longer duration of treatment is not accompanied by the appearance of new and/or more severe side-effects. PMID- 9352417 TI - The in vitro effect of hydroxychloroquine on skin morphology and transglutaminase. AB - BACKGROUND: Antimalarials are some of the most notorious drugs which may induce psoriasis, with 25% of all reported cases being associated with them. Antimalarials do not induce psoriasis de novo, but trigger subclinical psoriasis. In a previous report, we suggested that antimalarials exert their effect by interfering with the epidermal transglutaminase (TGase) activity. OBJECTIVE: To verify this hypothesis by examining the effect of hydroxychloroquine sulfate (HCQS) on cultured human skin and on TGase activity in vitro. MATERIALS AND METHODS: Skin samples from normal donors were cultured in the presence of HCQS for 4 days, and then processed for microscopic examination. TGase activity was assayed in the presence of HCQS and compared with blanks. RESULTS: Significant changes in epidermal morphology were seen in all explants cultured in the presence of HCQS at all concentrations employed. Areas of enhanced and irregular keratinization were observed in the upper epidermis, while a loss of cell polarity, with keratinocyte crowding and disarray, was seen in the lower epidermis. In addition, we observed intraepidermal splitting at different levels and dermo-epidermal detachments. HCQS showed a concentration-dependent inhibition of TGase activity. CONCLUSIONS: We suggest that HCQS causes an initial break in the barrier function of the epidermis by inhibiting TGase activity; this is followed by a physiologic response of the epidermis aimed at barrier restoration. This rather nonspecific stimulus to epidermal proliferation is probably sufficient to trigger psoriasis in predisposed individuals or aggravate it in psoriatic patients. PMID- 9352418 TI - Autoimmune progesterone dermatitis: effective prophylactic treatment with danazol. AB - BACKGROUND: Autoimmune progesterone dermatitis is a rare condition appearing during the perimenstrual period or following progesterone treatment. Various treatment modalities have been suggested, but most have proved to be ineffective. METHODS: We used the anabolic androgen danazol as a preventive treatment for recurrent episodes of autoimmune progesterone dermatitis in two young women. The treatment regimen consisted of 200 mg danazol twice daily, starting 1-2 days before the expected date of each menses and continuing for 3 days thereafter. RESULTS: This treatment regimen proved to be highly effective in preventing the eruptions in these two patients. CONCLUSIONS: Patients with autoimmune progesterone dermatitis may benefit from prophylactic treatment with danazol. PMID- 9352419 TI - The "anthrax" of two Byzantine emperors: Constantine V (741-775) and Leo IV (775 780). PMID- 9352420 TI - Herpes zoster-like Sweet's syndrome in acute myelogenous leukemia. PMID- 9352421 TI - Epinephrine-induced ischemia. PMID- 9352422 TI - Topical cyclosporine-A for treatment of oral ulcers of Behcet's syndrome. PMID- 9352424 TI - Reversal of a visuoconstructional disorder by weak electromagnetic fields in a child with Tourette's syndrome. AB - Tourette's syndrome (TS), a chronic familial neuropsychiatric disorder of unknown etiology, is characterized clinically by the occurrence of motor and vocal tics and by the presence of a variety of neurobehavioral and neurocognitive abnormalities including hyperactivity, self-mutilatory behavior, obsessive compulsive behavior, learning disabilities, and conduct disorder. Cognitive deficits related to right hemispheric dysfunction are common in TS patients accounting for decrements in visuospatial, visuoconstructional and visuomotor skills. An 11 year old boy with a 5 years history of TS exhibited during a routine neuropsychological assessment an unusual visuoconstructional disorder which previously has been observed in dyslexic children. Specifically, when instructed to draw a bicycle from memory, he drew spontaneously a design executed from the perspective of a bird's eye view. After receiving a 20 minute treatment session with picotesla range electromagnetic fields (EMFs) applied extracranially, this visuocontructional disorder was spontaneously reversed and he drew an elaborate and detailed bicycle positioned in profile. A placebo EMF treatment, which was administered prior to magnetic therapy, had no effect on this child's visuoconstructional disorder. During the ensuing week there was a marked reduction in the child's hyperactive behavior with attentuation of motor tics. Spontaneous drawing of a bicycle a week after the administration of magnetic therapy was executed in profile although some elements were presented from a bird's eye view. This case demonstrates the potential impact of treatment with picotesla EMFs in reversing specific cognitive deficits in TS related to right posterior hemispheric dysfunction. PMID- 9352423 TI - Resolution of sleep paralysis by weak electromagnetic fields in a patient with multiple sclerosis. AB - Sleep paralysis refers to episodes of inability to move during the onset of sleep or more commonly upon awakening. Patients often describe the sensation of struggling to move and may experience simultaneous frightening vivid hallucinations and dreams. Sleep paralysis and other manifestations of dissociated states of wakefulness and sleep, which reflect deficient monoaminergic regulation of neural modulators of REM sleep, have been reported in patients with multiple sclerosis (MS). A 40 year old woman with remitting progressive multiple sclerosis (MS) experienced episodes of sleep paralysis since the age of 16, four years prior to the onset of her neurological symptoms. Episodes of sleep paralysis, which manifested at a frequency of about once a week, occurred only upon awakening in the morning and were considered by the patient as a most terrifying experience. Periods of mental stress, sleep deprivation, physical fatigue and exacerbation of MS symptoms appeared to enhance the occurrence of sleep paralysis. In July of 1992 the patient began experimental treatment with AC pulsed applications of picotesla intensity electromagnetic fields (EMFs) of 5Hz frequency which were applied extracerebrally 1-2 times per week. During the course of treatment with EMFs the patient made a dramatic recovery of symptoms with improvement in vision, mobility, balance, bladder control, fatigue and short term memory. In addition, her baseline pattern reversal visual evoked potential studies, which showed abnormally prolonged latencies in both eyes, normalized 3 weeks after the initiation of magnetic therapy and remained normal more than 2.5 years later. Since the introduction of magnetic therapy episodes of sleep paralysis gradually diminished and abated completely over the past 3 years. This report suggests that MS may be associated with deficient REM sleep inhibitory neural mechanisms leading to sleep paralysis secondary to the intrusion of REM sleep atonia and dream imagery into the waking state. Pineal melatonin and monoaminergic neurons have been implicated in the induction and maintenance of REM sleep and the pathogenesis of sleep paralysis and it is suggested that resolution of sleep paralysis in this patient by AC pulsed applications of EMFs was related to enhancement of melatonin circadian rhythms and cerebral serotoninergic neurotransmission. PMID- 9352425 TI - Modulation by calcium of Deiters' neuron GABAA receptors in the rabbit. AB - The function of classical GABAA receptors of the rabbit Deiters' neurons has been studied at the single membrane level by a biochemical micromethod involving the study of labelled chloride permeation. In particular, labelled chloride permeation across microdissected fresh single membranes was studied in a microchamber system. The stimulation of 36Cl- out-->in permeation by "extracellular" GABA was determined under different conditions in the respect of Ca++. When the conditions were such that "intracellular" Ca++ was 0.02 microM there appeared to be an optimal effect by GABA on chloride passage. Conditions presumably resulting in an increase of [Ca++]i beyond the level reported above led to a decreased GABA effect, especially at the highest GABA concentrations used (> or = 10(-4)M). However, complete removal of Ca++ by a high (12 mM) intracellular EGTA concentration erased completely the GABA effect. These results indicate that in these neurons an optimal GABAA receptor function requires [Ca++]i levels well below micromolar. The high [EGTA]i effect seems to imply that too low a [Ca++]i is also harmful to the proper function of these GABAA receptors. However, an alternative explanation is possible. PMID- 9352426 TI - Treatment with electromagnetic fields reverses the long-term clinical course of a patient with chronic progressive multiple sclerosis. AB - It is estimated that 10-20% of patients with multiple sclerosis (MS) have a chronic progressive (CP) course characterized by an insidious onset of neurological deficits followed by steady progression of disability in the absence of symptomatic remission. To date no therapeutic modality has proven effective in reversing the clinical course of CP MS although there are indications that prolonged treatment with picotesla electromagnetic fields (EMFs) alters the clinical course of patients with CP MS. A 40 year-old woman presented in December of 1992 with CP MS with symptoms of spastic paraplegia, loss of trunk control, marked weakness of the upper limbs with loss of fine and gross motor hand functions, severe fatigue, cognitive deficits, mental depression, and autonomic dysfunction with neurogenic bladder and bowel incontinence. Her symptoms began at the age of 18 with weakness of the right leg and fatigue with long distance walking and over the ensuing years she experienced steady deterioration of functions. In 1985 she became wheelchair dependent and it was anticipated that within 1-2 years she would become functionally quadriplegic. In December of 1992 she began experimental treatment with EMFs. While receiving regularly weekly transcortical treatments with AC pulsed EMFs in the picotesla range intensity she experienced during the first year improvement in mental functions, return of strength in the upper extremities, and recovery of trunk control. During the second year she experienced the return of more hip functions and recovery of motor functions began in her legs. For the first time in years she can now initiate dorsiflexion of her ankles and actively extend her knees voluntarily. Over the past year she started to show signs of redevelopment of reciprocal gait. Presently, with enough function restored in her legs, she is learning to walk with a walker and is able to stand unassisted and maintain her balance for a few minutes. She also regained about 80% of functions in the upper limbs and hands. Most remarkably, there was no further progression of the disease during the 4 years course of magnetic therapy. This patient's clinical recovery cannot be explained on the basis of a spontaneous remission. It is suggested that pulsed applications of picotesla EMFs affect the neurobiological and immunological mechanisms underlying the pathogenesis of CP MS. PMID- 9352427 TI - Increased in vitro induced CD4+ and CD8+ T cell IFN-gamma and CD4+ T cell IL-10 production in stable relapsing multiple sclerosis. AB - Multiple sclerosis (MS) is presumed to be a T-cell mediated chronic inflammatory disease of the central nervous system. Investigators previously demonstrated increased IFN-gamma (pro-inflammatory) and IL-10 (counterregulatory anti inflammatory) in MS. The balance of pro-inflammatory and counterregulatory anti inflammatory cytokines may be important in the stabilization of disease activity. Purified CD4+ and CD8+ T cells from patients with clinically definite, stable relapsing MS (RRMS) were stimulated by anti-CD3 mAb or Con A for 48 hours and cytokine supernatants analysed for production of IL-2, IL-6, IFN-gamma, TNF-alpha (potential pro-inflammatory) and IL-4, IL-10, and TGF-beta (potential counterregulatory anti-inflammatory). Con A activated CD4+ and CD8+ T cell proinflammatory cytokine IL-2 secretion, CD4+ T cell IL-6 secretion, CD4+ and CD8+ T cell TNF-alpha secretion and CD8+ T cell IFN-gamma secretion was decreased significantly in RRMS subjects compared to controls. CD3 activated CD4+ and CD8+ T cell IL-6 secretion and CD4+ T cell TNF-alpha secretion was significantly decreased in MS subjects compared to controls. In contrast, there was increased CD3-induced IFN-gamma in both CD4+ and CD8+ T cells and counterregulatory anti inflammatory CD3-induced IL-10 secretion in CD4+ T cells in RRMS compared to controls. These data suggest that an equilibrium of a pro-inflammatory (IFN gamma) and a counterregulatory anti-inflammatory (IL-10) cytokine may define stable clinically definite early RRMS. PMID- 9352428 TI - Astrocytes grafted into rat nucleus basalis magnocellularis immediately after ibotenic acid injection fail to survive and have no effect on functional recovery. AB - In order to determine if the "trophic" properties of astrocytes makes them appropriate for use as a therapeutic agent to excitotoxic brain damage, adult male rats received grafts of cultured cerebral cortical astrocytes into the NBM immediately after infusion of ibotenic acid into the same structure. Twenty four hours after grafting and every other day for 11 days post surgery, the animals were tested for locomotor activity and habituation in an open field. Animals with NBM lesions had significantly reduced rearing activity as compared to counterparts with no lesions. Nine days after surgery, rats with NBM lesions and astrocyte grafts were as impaired in the acquisition of passive avoidance (PA) as their untreated counterparts. All animals with ibotenic lesions were impaired on PA retention compared to rats with no lesions. There was no difference between animals that had received grafts and those that had not. Fourteen days after grafting, all brains were processed for Nissl stain, acetylcholinesterase (AChE) histochemistry, GFAP immunocytochemistry, and bisbenzamide fluorescent microscopy. Decreases in the number of neurons in the NBM as well as decreases in the density of AChE staining in the ipsilateral cortex (the area of innervation of the NBM cholinergic neurons) was evident in all animals with NBM lesions. In addition, a large number of host reactive astrocytes were seen within the NBM, its vicinity, and in the ipsilateral neocortex. Grafted astrocytes survived and integrated into the host tissue when they were grafted into the brain of intact animals but no living grafted astrocytes were found in animals injected with ibotenate. In this latter case, two weeks after grafting, instead of surviving astrocytes only fluorescent tissue 'masses' were seen in the NBM, surrounded by a cavity. Grafted astrocytes did not have any effect on the extension of the lesion caused by ibotenic acid infusion. These results suggest that the concentration of ibotenic acid used to injure the NBM killed not only the host cholinergic neurons but also the grafted astrocytes. The failure of astrocytes to ameliorate the behavioral deficits caused by ibotenic acid lesions of the NBM may be due to the ibotenic acid creating a lethal environment for the grafted and freshly dissociated, cultured astrocytes. PMID- 9352429 TI - The anticonvulsant effect of deprenyl on pentylenetetrazol-induced seizures in Lewis rats. AB - There is recent evidence that deprenyl may have anticonvulsant action in a rat kindling model of epilepsy as well as in a maximal electroshock model. We therefore investigated the effect of deprenyl on the brain sensitivity threshold to pentylenetetrazol (PTZ)-induced maximal seizures in Lewis rats, in a model that provides pharmacodynamic information free of pharmacokinetic interference. The novel finding of this investigation was the anticonvulsant effect of deprenyl following repetitive administration whereas a single deprenyl dose did not affect the PTZ concentrations required to induce maximal seizures. The data suggests that the mechanism of this effect is not associated with the dopaminergic activity of deprenyl since pretreatment with both bromocriptine (a dopamine D2 agonist) and haloperidol (dopamine antagonist) did not affect the seizure threshold, whereas levodopa caused a proconvulsant effect. It was also concluded that the mechanism is not related to changes in acetylcholine levels since prolonged pretreatment with deprenyl did not attenuate the brain sensitivity to pilocarpine-induced seizures. The fact that long term administration of deprenyl was needed to produce its anticonvulsant effect may indicate that the anticonvulsant effect of deprenyl may be due to changes in levels of certain endogenous compounds or down or up-regulation of relevant receptor/effector units. PMID- 9352430 TI - Event-related potentials in a two-choice task involving within-form comparisons of pictures and words. AB - Behavioral as well as electrophysiological evidence suggests that words are processed differently than pictures in a variety of tasks. In this study fifteen adult subjects were tested on a speeded "same-different" judgment task between printed names and drawings of common objects. In one condition, subjects decided on the identity between their internal image of the object that was named by S1 (word) and a subsequently presented drawing of an object (Word-Picture condition). In a second condition, comparisons were made on the basis of the name of the depicted object (Picture-Word trials). Event-related potentials (ERPs) were recorded from 12 scalp locations in response to the second item in each pair. No-match waveforms were characterized by larger N350 and P500 deflections compared to Match ERPs. The two responses could be distinguished on the basis of their lateral and anterior-posterior scalp distribution. Within-form comparisons involving pictures produced increased positivity between 150 and 600 ms poststimulus onset at posterior recording sites, whereas the opposite effect was noted at anterior sites during the early portion of the ERP. Decisions on word stimuli were associated with prolonged reaction time and longer N350 peak latency compared to decisions on pictures. These results demonstrate the existence of independent sources of ERP variability, each possibly reflecting a different aspect of cognitive comparisons. Latency and reaction time data provided valuable information regarding differences in the course of the comparison process when linguistic, as opposed to pictorial, stimuli/representations were involved. PMID- 9352431 TI - Is the hippocampus involved in temporal discrimination and the memory of short intervals? AB - Rats with lesions to the hippocampus proper and the subiculum were tested for timing behavior and temporal memory. Using the peak procedure, they were trained to discriminate a 40 s interval and a retention gap tested the memory for time. Results were interpreted within the theoretical framework of the internal clock and with respect to current theories on hippocampal function. Timing behavior was unaffected by either lesion and no shifts in the temporal discrimination functions were observed. The lesions also failed to show a deficit in the memory for temporal events. For all groups, the retention gap increased the mean peak time by the time of the gap. This indicated that all rats used the stop rule which required the use of working memory. Thus, it was concluded that the hippocampus is neither necessary for accurate timing behavior nor for the memory of temporal events. PMID- 9352432 TI - The accelerated aging hypothesis of Parkinson's disease is not supported by the pattern of circadian melatonin secretion. AB - Hypotheses pertaining to the etiology of Parkinson's disease (PD) have suggested that the disease reflects an accelerated form of the normal aging process. Aging is associated with progressive failure of the pineal gland associated with a gradual decline in nighttime plasma melatonin secretion. The decline in melatonin secretion with age, at an average rate of 10-15% per decade, is considered a marker of brain aging in humans and estimations of plasma melatonin levels could be used to distinguish the processes of normal aging from pathological age related changes. The accelerated aging hypothesis of PD is not supported by studies which have examined nocturnal melatonin secretion in drug naive Parkinsonian patients compared to age matched normal control subjects. Specifically, these studies have revealed no significant differences in the melatonin rhythms (i.e., peak nocturnal melatonin level and 24-hour melatonin output) between PD patients and normal age matched controls. On the other hand, melatonin secretion is significantly lower in Alzheimer's patients compared to age matched normal subjects. Collectively, it is suggested, on the basis of melatonin circadian rhythms, that Alzheimer's disease rather than PD is related to an accelerated aging process, a hypothesis which is supported by pathological and neurochemical studies. PMID- 9352433 TI - Aging and olfactory recognition memory: effect of encoding strategies and cognitive abilities. AB - The effects of two encoding manipulations on recognition memory for odors were examined in 20 young and 20 elderly males. Subjects were instructed to use two different encoding strategies: (1) labeling-plus-definition (i.e., naming the odor and giving a short description) and, (2) life-episode association (i.e., associating a memory of a life-episode with each odor). Results revealed that elderly subjects performed significantly worse than young subjects in the labeling-plus-definition condition, but not in the life-episode task in which they achieved a level of performance not significantly different from the young. Encoding specificity or precision did not significantly impact recognition memory for the odors in either group. Analysis of response bias revealed that, in the label-plus-definition condition, young subjects were more conservative in responding style, while more liberal in their responding in the life-episode condition. In contrast, elderly subjects exhibited nearly identical response bias across both encoding conditions. The results suggest that age-related deficits in olfactory memory may be strategy dependent. PMID- 9352434 TI - Prognostic value of nuclear morphometry in feline mammary carcinomas. AB - This study of 30 cats with mammary carcinoma was designed to investigate the relationship between (1) six nuclear morphometric parameters (mean nuclear profile area [MNA], standard deviation of MNA [SDa], coefficient of variation of MNA [CVa], nuclear form factor (p2/4 pi area) [FF], standard deviation of FF [SDf], and coefficient of variation of FF [CVf]) assessed by image analysis, and (2) survival for > 1 year or < 1 year after surgical removal of the tumour. Only the SDf and the CVf appeared to be related to survival. Cats that died within 1 year had an SDf or CVf (or both) higher than the corresponding mean values (SDft and CVft) for all 30 cats; but only four of 16 cases (25%) with a SDf lower than SDft and five of 17 cases (29.4%) with a CVf lower than CVft died within 1 year. The authors conclude that SDf and CVf represent reliable prognostic parameters in feline mammary carcinomas. PMID- 9352435 TI - Non-suppurative myocarditis in piglets associated with porcine parvovirus infection. AB - The involvement of porcine parvovirus (PPV) in the aetiology of non-suppurative myocarditis in sucking piglets was investigated by a polymerase chain reaction (PCR), designed to assess the presence of viral genome in formalin-fixed paraffin wax-embedded tissue of diseased animals. Myocardium and lung of stillborn piglets with a confirmed PPV infection were used to set up the PCR amplification method. Subsequently, 20 myocardia with inflammatory lesions were examined in parallel with 20 myocardia without lesions, from age-matched control piglets. Tissues were first tested for the presence and the integrity of porcine DNA by amplifying a sequence encoding the highly conserved nuclear protein histone H4. Tissue from 15 out of 20 animals with myocarditis contained amplifiable histone H4 DNA and in 12 of the 15 histone H4-positive samples, PPV DNA was detected. It proved possible to amplify histone H4 DNA in all 20 negative controls (without myocarditis), and PPV DNA was detected in three cases. In-situ hybridization with a digoxigenin labelled probe homologous to PPV was performed in four PCR-positive cases of non suppurative myocarditis. In two animals several positively stained nuclei were observed in the myocardium, within or close to the mild inflammatory cellular infiltrates. These results strongly suggest that PPV can cause non-suppurative myocarditis in sucking piglets. PMID- 9352436 TI - Protective effect of exposure to non-virulent Trypanosoma cruzi clones on the course of subsequent infections with highly virulent clones in mice. AB - The protective effect of primary infection with non-virulent Trypanosoma cruzi clones against subsequent infection with highly virulent clones was determined in groups of BALB/c mice. All mice inoculated with the highly virulent m3 and m4 clones succumbed in < or = 16 days. Mice inoculated with the non-virulent h1 and h2 clones survived and were superinfected with the m3 and m4 clones. Low degrees of parasitaemia were observed in mice challenged with the highly virulent clones. The survival ratios of the superinfected mice were not statistically different from those seen in mice that received a single injection of non-virulent T. cruzi. Mice given a non-virulent infection and subsequently challenged with a virulent clone differed from those given only a non-virulent infection in showing more frequently an inflammatory infiltrate in the heart, skeletal muscle and intestines. PMID- 9352437 TI - Histological and immunohistochemical findings in thoracic lymph nodes of cattle with contagious bovine pleuropneumonia. AB - Outbreaks of contagious bovine pleuropneumonia (CBPP) were reported in Lombardy, Northern Italy, at the end of 1990. For the purpose of this study, 54 slaughtered Holstein-Friesian cows showing typical lung lesions of CBPP from which the small colony type of Mycoplasma mycoides subspecies mycoides (M. m. mycoides SC) was isolated, were selected. Thoracic lymph nodes from these animals were sampled for bacteriological, histological and immunohistochemical analysis. Acute, subacute and chronic lesions were observed in 13, 12 and 29 cases, respectively. In the 13 animals showing acute lung lesions, an increased number of macrophages was observed, especially in the subcapsular sinuses, but frequently also in the cortical and medullary sinuses of the thoracic lymph nodes; in all 13 acute cases M. m. mycoides SC antigen was detected immunohistochemically in the cytoplasm of the macrophages. In 10 out of the 12 cases with subacute lung lesions, mycoplasma antigen was observed in macrophages located in sinuses, as well as in those scattered in the lymph node parenchyma. Hyperplasia of germinal centres in follicles was observed histologically in most of the 29 cases with chronic lung lesions. In immunohistochemically labelled sections, the characteristic finding observed in 27 of the chronic cases, was the presence of a variable amount of positive material in the germinal centres. These findings demonstrate the involvement of thoracic lymph nodes in CBPP. PMID- 9352438 TI - Influence of immunization on the pulmonary inflammatory response of rabbits induced by Pasteurella haemolytica A1 lipopolysaccharide. AB - Immune complex formation has long been thought to play a role in the pathogenesis of Pasteurella haemolytica pneumonia. This study in laboratory rabbits was designed to investigate immune-mediated damage in respiratory tissue caused by lipopolysaccharide (LPS). Severe lesions were induced by the intratracheal (IT) injection of P. haemolytica A1 LPS (50 micrograms) into rabbits previously immunized with P. haemolytica killed whole cells emulsified with Freund's incomplete adjuvant (FIA); these lesions included perivascular oedema and polymorphonuclear leucocyte (PMN) infiltration of the subintima, with degeneration and necrosis of the media. Smaller vessels were occluded by PMNs in various stages of degranulation. PMN counts in bronchoalveolar lavage (BAL) fluid were significantly elevated (P < 0.05). Lesions were also induced by the IT injection of LPS (50 micrograms) into rabbits pretreated with an emulsion consisting merely of FIA and formol-saline; these lesions included moderate to severe congestion, interstitial oedema, alveolar serofibrinous exudation and PMN infiltration. PMNs were also present in BAL fluid. Rabbits pretreated with FIA in formol-saline and given a later IT injection of saline, and rabbits pretreated with bovine serum albumin (BSA) in FIA and given a later IT injection of BSA, were included as negative and positive control groups. Cutaneous lesions were also induced by the intradermal injection of LPS into rabbits immunized against P. haemolytica and of BSA into rabbits immunized with BSA. Overall, the pulmonary and cutaneous lesions induced in vaccinated rabbits by antigen administration were more severe than those seen in non-vaccinated rabbits. The lesions in rabbits, which were similar to those seen in natural cases of P. haemolytica pneumonia in cattle, were characterized by a fibrinopurulent inflammatory process with extensive interstitial oedema, fibrinous exudate, and PMNs. This model may help to elucidate the pathogenesis of pneumonic pasteurellosis in immunized animals. PMID- 9352439 TI - Disseminated intravascular coagulation in chickens inoculated with Erysipelothrix rhusiopathiae. AB - In a first experiment, 28 specific pathogen-free chickens aged 3 weeks showed clinical signs 1 to 5 days after intramuscular inoculation with Erysipelothrix rhusiopathiae. Twelve of 28 birds died 2 to 4 days after inoculation. Macroscopically, the liver, spleen and kidneys were seen to be enlarged and congested. Histologically, fibrinous thrombus formation, seen in the hepatic sinusoids, renal glomerular capillaries and small pulmonary blood vessels, was a characteristic feature. In addition, the liver showed marked congestion, increase of mononuclear cells and heterophils in the sinusoids, hyperplasia of sinusoidal lining cells, and vacuolar changes in hepatic cells. The spleen showed fibrinous exudation of the lymphoid follicles and ellipsoids with lymphocytic depletion, and hyperplasia of ellipsoidal reticular cells. There was oedema, congestion and cellular infiltration in the interstitium of the kidney. The bursa of Fabricius and thymus showed marked lymphocytic depletion. In a second experiment, the blood chemical values (uric acid, glutamic-oxalacetic transaminase, lactate dehydrogenase and gamma-glutamyl transpeptidase) of birds inoculated intramuscularly with E. rhusiopathiae were significantly higher than those of uninfected controls. The blood prothrombin times and activated partial thromboplastin times of the inoculated group were significantly greater than those of the control group. The pathological and haematological findings demonstrated that E. rhusiopathiae induced disseminated intravascular coagulation in the chickens. PMID- 9352440 TI - Detection of nucleic acids of porcine reproductive and respiratory syndrome virus in the lungs of naturally infected piglets as determined by in-situ hybridization. AB - Replication of porcine reproductive and respiratory syndrome virus (PRRSV) was studied in formalin-fixed paraffin wax-embedded lung tissues from seven naturally infected piglets by in-situ hybridization with a non-radioactive digoxigenin labelled probe. A 433 base pair cDNA probe for the viral RNA encoding the nucleocapsid proteins of a Korean PRRSV isolate was generated by the polymerase chain reaction. All seven piglets infected with PRRSV showed a distinct, positive signal, scattered throughout the alveolar septa and spaces. Positive cells typically exhibited dark brown staining deposits in the cytoplasm without background staining. In-situ hybridization demonstrated that PRRSV replicated primarily in interstitial and alveolar macrophages, and occasionally in type 2 pneumocytes. The bronchial or bronchiolar epithelium did not exhibit a hybridization signal for PRRSV nucleic acids. The anterior and middle lobes of the lung were more reliable than the caudal or accessory lobes for the detection of PRRSV nucleic acids. The in-situ hybridization technique used was rapid, specific and sensitive, and may prove useful for the diagnosis of PRRSV infection in routinely fixed and processed tissues. PMID- 9352441 TI - Adaptation of a fluorocarbon-based non-aqueous fixation regime for the ultrastructural study of the teleost epithelial mucous coat. AB - Studies on the microanatomy of the mucus-rich biofilm surface of normal or damaged teleost skin tissue have been limited because conventional fixation regimes do not effectively retain mucus during tissue preparation. A non-aqueous fixation method, based on a technique devised to retain airway mucous for ultrastructural study, and consisting of the use of an inert perfluorocarbon solvent with osmium teroxide 1%, was successfully used to prepare skin tissues of healthy juvenile rainbow trout. The skin's mucous coat was examined by transmission electron microscopy and the results were compared with those obtained with tissues prepared by a conventional glutaraldehyde-based method. In samples fixed with glutaraldehyde, the cell-surface structures retained were limited to microridges and a poorly discernible glycocalyx layer. In contrast, those fixed by the non-aqueous method had a more clearly demonstrated glycocalyx layer, and a second fibrillar layer, resembling mucus, which was separated from the glycocalyx layer by an electron-lucent zone. PMID- 9352442 TI - Pituitary adenoma with prolactin and growth hormone production in a sheep. AB - A pituitary adenoma was found in a 6-year-old ewe. This tumour was composed mainly of chromophobic cells, but acidophilic cells predominated in a few areas. Prolactin was demonstrated in the cytoplasm of some chromophobic cells by means of the immunoperoxidase technique, and secretory granules ranging in size from 100 to 900 nm in diameter were seen in all chromophobic cells examined. Almost all acidophilic cells were positive for growth hormone (GH), and a few of them were also positive for prolactin (PRL). This tumour resembled a human mixed GH cell-PRL cell adenoma. PMID- 9352443 TI - Immunohistological assessment of fibrin deposition and thrombus formation in canine mammary neoplasia. AB - A commercially available monoclonal antibody against human fibrin was used to detect fibrin in canine formalin-fixed, paraffin wax-embedded tissue by applying a slightly modified alkaline phosphatase anti-alkaline phosphatase (APAAP) technique. Twenty-eight mammary tumours from six bitches were examined for the presence of fibrin. Thrombi and extravascular fibrin deposits were detected in 15 tumours (12 complex adenocarcinomas, one adenocarcinoma, two solid carcinomas), and a single thrombus was detected in one adenoma; 12 tumours (three adenomas, one complex adenoma, four complex adenocarcinomas and four adenocarcinomas) did not show any staining reaction. PMID- 9352444 TI - Segmented filamentous bacteria in the bovine small intestine. AB - Segmented filamentous bacteria (SFB) were observed in a 28-day-old calf, attached to the absorptive villi. Morphologically, they were similar to SFB described in other animal species. Because these organisms cannot be cultured, further characterization was not possible. The organisms were confined to the upper third of the absorptive villi and were not seen attached to the follicle-associated epithelium of the Peyer's patch, or observed in the caecum or colon. Although they were often associated with minor lesions, their pathological significance was doubtful. With this report, segmented filamentous bacteria have now been described in virtually all the commercially important livestock and poultry species, in other domestic animals, and in man. PMID- 9352445 TI - Autonomic synaptic transmission at single boutons and calyces. PMID- 9352446 TI - Delayed rhodopsin regeneration and altered distribution of interphotoreceptor retinoid binding protein (IRBP) in the mi(vit)/mi(vit) (vitiligo) mouse. AB - Rhodopsin regeneration requires attachment between the retinal pigment epithelium (RPE) and rod outer segments; however, in experimentally induced retinal detachment, rhodopsin regeneration can be restored partially upon addition of IRBP (interphotoreceptor retinoid binding protein). The mi(vit)/mi(vit) (vitiligo) mutant mouse, a model of slowly progressing photoreceptor cell degeneration, has a marked elevation of IRBP at 4 weeks as well as progressive detachment of the retina. The purpose of this study was to determine whether this mutant is capable of regenerating rhodopsin within a few hours following an intense light bleach. Rhodopsin regeneration was determined spectrophotometrically in mice after an intense one hour light bleach followed by 0,1,2,4 or 24 h of dark recovery. IRBP was localized immunohistochemically in fixed frozen tissue at the light microscopic level and in LR Gold embedded tissue at the ultrastructural level. Rhodopsin regeneration experiments indicated that rhodopsin levels following 0,1,2 and 4 h dark-recovery were significantly less in mi(vit)/mi(vit) mutants compared with controls. Immunohistochemical detection of IRBP indicated an altered distribution of the protein in the mutant mice compared with controls. There was accumulation in the region of the inner segments in mutant retinas rather than distribution only to the RPE/OS apical regions as in controls. The data suggest that regeneration of rhodopsin is reduced by 4 weeks postnatally in the mi(vit)/mi(vit) mouse. There is partial detachment of the retina at this age; and IRBP, thought to be essential for proper functioning of the visual cycle, is aberrantly distributed in this mutant. PMID- 9352447 TI - Axotomy-induced apoptosis in adult rat primary sensory neurons. AB - Neuronal death following unilateral axotomy of a sensory nerve has long been inferred from neuronal counts of dorsal root ganglion neurons, using the contralateral ganglia as a control. The counting methods used usually involved the counting of neuronal nucleoli and made assumptions about them which could conceivably be flawed. Very few studies have used direct observations of dying or degenerating neurons to address questions concerning the duration of the period of neuronal death or the mechanisms involved in this process. Here we describe a morphological, morphometric and histochemical study into the nature and duration of sensory neuron death following transection and ligation of the sciatic nerve at mid-thigh level in the adult rat. We show that at least some of this neuronal loss occurs by apoptosis as defined by morphological criteria and in situ end labelling of damaged DNA. Absolute numbers of apoptotic neurons were counted from serial paraffin sections of ganglia and estimates of neuronal numbers obtained by disector analysis at 1, 2, 3 and 6 months after axotomy. Using this approach we show that axotomy-induced apoptosis begins at around 1 week and continues up to at least 6 months after axotomy. PMID- 9352448 TI - Spongiform degeneration of the gerbil cochlear nucleus: an ultrastructural and immunohistochemical evaluation. AB - Observations from ultrastructural and immunohistochemical studies suggest that spongiform lesions in the gerbil cochlear nucleus are derived principally from dendrites. Almost one-fifth of the lesion profiles examined ultrastructurally exhibited synaptic contacts with axon terminals. In addition, approximately 80% of lesions are immunopositive for the dendrite-specific microtubule associated protein, MAP2. Ultrastructural studies showed a small percentage (8%) of lesions were derived from myelinated axons, although none were immunohistochemically labelled with antibodies to the tau protein. Staining with the astrocyte-specific markers GFAP, S-100 and vimentin yielded equivocal results, but did not support a major role for astrocytes in lesion formation. The histological profile matches that seen in some other well characterized types of spongiform degeneration. PMID- 9352449 TI - Mitral annulus calcification is not an independent risk factor for stroke: a cohort study of 657 patients. AB - All studies but one in the past have shown a strong relative risk of mitral annulus calcification for stroke, but the contribution of associated cardiac and vascular risk factors, especially carotid atheroma has not been appreciated. We studied the risk of stroke in selected patients with mitral annular calcification, adjusting for clinical, echocardiographic and therapeutic factors influencing stroke risk. Of 8,160 consecutive patients with echocardiograms, 657 with and 562 without mitral annulus calcification were followed for a mean of 2.4 years (range 1-6.6) to determine stroke risk by means of proportional hazards models with clinical, echocardiographic, and therapeutic variables that influence the risk of stroke. We also determined the association of mitral annulus calcification with subtypes of ischaemic brain lesions generally considered to be specific for an underlying cardioembolic cause. We therefore distinguished between territorial, small deep, and asymptomatic (silent) brain infarcts. Fifty one patients with mitral annulus calcification and 27 controls had a stroke in the follow-up period. Mitral annulus calcification was not significantly associated with stroke in proportional hazards models (hazard ratio 0.76, 95% confidence interval 0.42-1.36, P = 0.3), or with any of the stroke subtypes, or with the presence of silent brain infarcts after adjustments for risk factors for generalized vascular disease Hypertension and carotid atheroma, with or without stenosis, ipsilateral or contralateral to the side of the stroke, were significantly associated with stroke in our patients. This study does not support the view that mitral annulus calcification is a risk factor for stroke. As others have found strong associations between mitral annulus calcification and cardiac and vascular risk factors for stroke, the increased risk of stroke in patients with mitral annulus calcification reported may be explained by these confounding risk factors. Therefore, in our opinion, mitral annulus calcification requires treatment of cardiovascular risk factors, but generally no specific measures such as surgery or oral anticoagulants are required to lower the risk of stroke. PMID- 9352450 TI - Long-term cyclosporine treatment in a group of severe myasthenia gravis patients. AB - We evaluated cyclosporine A (CsA) treatment in 9 patients (6 female and 3 male), 16-63 years old, with severe myasthenia gravis (MG) for a mean period of 2 years (range 16-36 months). All of the patients had been previously treated either with corticosteroids or by combined immunotherapy, and 5 needed periodic plasma exchanges. The reduction of plasmapheresis cycles in the 5 patients who needed periodic plasma exchange to maintain an acceptable quality of life showed an impressive cost-benefit analysis. During CsA treatment 7 of 9 patients improved their muscle strength and functional score. In all the patients except one the corticosteroid dosage was reduced and in 7 of the 9 patients the dose reduction was over 50% with subsequent reduction of the corticosteroid side effects. The findings showed that initiation of CsA treatment increased muscle strength and reduced corticosteroid dosage. The most common CsA side effects were: a serum creatinine increase that occurred in the first 6-12 months of therapy in 8 patients, other side effects like hypertrichosis and gingival hyperplasia were present in four patients. Blood pressure increase was found in only one patient. CsA treatment may be a valuable and cost effective treatment in severe MG. PMID- 9352451 TI - Prognosis of myasthenia gravis: a retrospective study of 380 patients. AB - The 9139 follow-up records of 438 myasthenia gravis (MG) patients were reviewed. Excluding those patients who were diagnosed 5 or more years after symptom onset (n = 37) and those who experienced only oculomotor symptoms throughout follow-up (n = 21), there were 380 patients. A survival analysis approach was used to assess the influence of prognostic factors on the following endpoints: (a) stable complete remission, (b) complete remission of at least 6 months and (c) pharmacological remission of at least 6 months. Early diagnosis was associated with a better prognosis with respect to all endpoints. Thymectomy also improved the prognosis but only for those patients without thymoma. Later MG onset was associated with a higher tendency to achieve pharmacological remission. PMID- 9352452 TI - The prognostic significance of coma-rating, duration of anoxia and cardiopulmonary resuscitation in out-of-hospital cardiac arrest. AB - Early determination of outcome after successful prehospital cardiopulmonary resuscitation (CPR) is a common problem with great ethical, economic, social, and legal consequences. We prospectively investigated 112 adult patients who had been resuscitated after out-of-hospital cardiac arrest (CA). The aim of our study was to determine whether coma rating by the mobile intensive care unit (MICU) is a useful tool for outcome prediction. For neurological assessment the Innsbruck Coma Scale (ICS) was used initially and after return of spontaneous circulation (ROSC) or 20-30 min after the start of CPR, before any sedating drugs were given. The duration of anoxia and CPR were determined with the automatically recorded emergency call protocol of the dispatch centre and the protocol of the MICU. For estimation of cerebral outcome at the time of discharge from hospital we used the Glasgow-Pittsburgh Cerebral Performance Categories (CPC). Restoration of spontaneous circulation was achieved in 42 patients (37%), and 15 (13%) were discharged from hospital. The first coma rating performed immediately at the time of arrival on scene had no significant prognostic value for prediction of neurological outcome (P = 0.204) and survival (P = 0.103). The second coma rating (performed after ROSC or 20-30 min after the start of CPR), however, demonstrated a significant correlation with neurological outcome (P = 0.0000) and survival (P = 0.0000), a correlation which was comparable to both duration of anoxia and duration of CPR. In patients with out-of-hospital cardiac arrest prognostic information could be obtained with the ICS as early as 20-30 min after the start of cardiopulmonary resuscitation. PMID- 9352453 TI - Hereditary motor and sensory neuropathy with spastic paraplegia and optic atrophy: report on a family. AB - We describe two siblings affected by a motor and sensory neuropathy starting in childhood. Already in infancy, a spastic gait disturbance had become obvious, leading later to multiple surgical interventions. In adolescence, progressive loss of vision developed. At the time of our examination, both siblings showed severe weakness and atrophy of the distal muscles of legs and arms. Tendon jerks were brisk in proximal muscles; in the lower extremities, muscle tone was increased. Visual acuity was severely decreased. Nerve conduction studies revealed an axonal degeneration. This finding was confirmed by evaluation of a sural biopsy specimen in one patient, showing only few remaining myelinated fibres without signs of demyelination. This combination of hereditary motor and sensory neuropathy with spastic paraplegia and optic atrophy shows features of both hereditary motor and sensory neuropathy V and VI according to the classification of Dyck, indicating that these subtypes may not represent distinct entities. PMID- 9352455 TI - Mechanisms of infarction in the superficial posterior cerebral artery territory. AB - Numerous reports have described a variety of clinical syndromes resulting from posterior cerebral artery (PCA) infarction, whereas only a few pathoanatomical and retrospective clinical studies have investigated the underlying mechanisms. Therefore we attempted to determine the causes of infarction in the superficial posterior cerebral artery (PCA) territory by means of a more comprehensive, modern vascular and cardiac study. During a 4-year period 74 consecutive patients (49 men, 25 women) with acute PCA infarction documented on CT (n = 74) and MRI (n = 41) were included in the study. Patients had a neurological examination, vascular studies [extra- and transcranial Doppler (n = 74), magnetic resonance (n = 31) or intra-arterial (n = 22) angiography], cardiac evaluation [ECG (n = 74), transthoracic (n = 74) and transoesophageal echocardiography (n = 30)], and coagulation tests. A cardiac source of embolism was established in 31%, significant vertebrobasilar artery disease in 22%, and PCA stenosis or occlusion in 8% of the patients. Rare causes, such as hypercoagulopathy or paradoxical embolism via a patent foramen ovale, were present in 15%. However, in spite of the comprehensive diagnostic evaluation, the cause of the stroke remained undetermined in 24% of the cases. Apart from complete infarcts of the posterior branches of the PCA, which occurred more frequently in cardioembolic strokes (18%, P < 0.05), the topographical patterns of infarct extension and the coincidence of infarction in the deep territories of the PCA, the cerebellum and brainstem were not significantly different among the causal subgroups. The frequency of haemorrhagic transformation (18%) was highest among cardioembolic strokes (44%, P < 0.001). This prospective study of PCA infarction demonstrated embolism from cardiac and vascular sources as the predominant cause. In contrast to previous studies, we found no evidence of migraine as a cause of PCA infarction, whereas paradoxical embolism was the presumed cause in a considerable number of cases. Whereas the cause of stroke could not reliably be derived from infarct topography, haemorrhagic transformation indicated there had been cardioembolism in most cases. PMID- 9352454 TI - S-100 protein concentration in the cerebrospinal fluid of patients with Creutzfeldt-Jakob disease. AB - We evaluated S-100 levels in paired cerebrospinal fluid (CSF) and serum samples in a group of 135 patients referred to the German Creutzfeldt-Jakob disease (CJD) surveillance unit from June 1993 to May 1995. The patients were seen in a prospective case control study. The diagnosis of probable CJD during life was made in any patient presenting with rapidly progressive dementia of less than 2 years' duration, typical periodic sharp wave complexes (PSWCs) in the EEG and at least two of the following findings: myoclonus, visual/or cerebellar symptoms, pyramidal and/or extrapyramidal signs and/or akinetic mutism. Patients presenting with the above clinical signs and symptoms but without PSWCs were classified as possible, while those with a dementia of a duration exceeding 2 years and without PSWCs were classified as other. S-100 was determined in paired CSF and serum samples by a commercially available enzyme-linked immunosorbent assay. In a group of 76 patients with definite and probable CJD, S-100 concentration (median 25 ng/ml, range 2-117) in CSF was significantly higher (P < 0.0001) than in 32 patients diagnosed as other (median 4 ng/ml, range 1-19). Serum levels of S-100 were below 0.5 ng/ml in all groups. At a cut-off of 8 ng/ml an optimum sensitivity of 84.2% with a specificity of 90.6% for the diagnosis of CJD by the determination of S-100 in CSF is obtained. S-100 levels exceeding 8 ng/ml in CSF support the diagnosis of CJD in any patient presenting with rapidly progressive dementia. PMID- 9352456 TI - Central motor conduction studies in internal capsule and corona radiata infarction. AB - Clinical and evoked-potential studies in internal capsule and corona radiata infarction are lacking. We report the results of a clinical and central motor conduction time (CMCT) study in 16 patients with internal capsule and 17 with computed tomography (CT)-proven corona radiata infarction. Patients's outcome was defined at the end of 3 months on the basis of the Barthel Index score. Four patients with type A capsular infarction (middle third of posterior limb of internal capsule) all had severe weakness, while 2 also had persistently unrecordable CMCT and poor outcome. Twelve patients with type B internal capsular infarction (genu, anterior limb, anterior or posterior third of posterior limb) had a milder degree of weakness, and CMCT was recordable in 9. At 3 months' follow-up, however, CMCT was recordable in all 12 patients. All of these patients had a partial (n = 4) or complete (n = 5) recovery. Thirteen patients with type A corona radiata infarction (middle third of corona radiata) had more pronounced weakness, and CMCT was unrecordable in all of these patients except 1 on initial examination. Follow-up after 3 months was possible in 8 patients, and CMCT became recordable in 3. One of these patients had complete, 3 partial, and 4 poor recovery. In type B corona radiata infarction (anterior or posterior third of corona radiata), the clinical signs and CMCT did not follow a regular pattern. Clinical and CMCT abnormalities in internal capsular infarction followed a more predictable pattern compared with those in corona radiata infarction. A less predictable pattern of weakness and CMCT change in corona radiata infarction may be attributed to a less definite organisation of motor pathways compared with the internal capsule. PMID- 9352457 TI - White matter lesion detection in multiple sclerosis: improved interobserver concordance with multispectral MRI display. AB - An assessment of the detectability of white matter lesions and of concordance between observers with different levels of MRI reading experience was performed with comparative evaluation of spin-echo MRI images and of corresponding "multispectral" maps in 16 patients with definite multiple sclerosis (MS). Multispectral maps were obtained by means of a recently described post-processing technique based on the simultaneous display of MRI parameters and a standardized colour scale with red, green and blue coding for relaxation rates R1 and R2 and proton density, respectively. Spin-echo images on films and multispectral maps displayed on a personal computer were randomly rated at 2-month intervals. Interobserver concordance (k-test) was assessed among three readers with different levels of MRI experience (an experienced neuroradiologist, a radiology resident and a neurologist). For multispectral maps we found increased interobserver concordance with the experienced neuroradiologist (multispectral vs conventional images; k = 0.77 vs 0.66 for the radiology resident and 0.66 vs 0.56 for the neurologist), an increased number of detected lesions and decreased reading time. Multispectral maps permit easy detection of MS lesions and may improve interobserver concordance compared with conventional spinecho studies. PMID- 9352459 TI - Infectious mononucleosis complicated by transverse myelitis: detection of the viral genome by polymerase chain reaction in the cerebrospinal fluid. PMID- 9352458 TI - Very high frequency of the His1069Gln mutation in Polish Wilson disease patients. PMID- 9352460 TI - A myotatic reflex of the extensor hallucis longus muscle. PMID- 9352461 TI - Adrenoleukodystrophy of very late onset. PMID- 9352462 TI - Potent in vivo inhibitors of rat renin: analogues of human and rat angiotensinogen sequences containing different classes of pseudodipeptides at the scissile site. AB - Using solid-phase methodology we have synthesised peptides based on the 8-14 or 6 14 human and rat angiotensinogen sequences, containing the following different isosteric units at the P1-P1' cleavage site: Leu-psi[CH2NH]Leu; Leu psi[CH(OH)CH2]Val; Leu-psi[CH(OH)CH2]Leu and Leu-psi[CH(NH2)CH2]Val. In vitro, peptide Piv-His-Pro-Phe-His-Leu-psi[CH(OH)CH2]Leu-Tyr-Tyr-Ser-NH2(XXI) is the most potent inhibitor of rat plasma renin reported having an IC50 of 0.21 nM; it is a much weaker inhibitor of human renin (IC50 45 nM). Peptide Boc-His-Pro-Phe His-Leu-psi[CH(OH)CH2] Leu-Val-Ile-His-NH2 (XX) was a highly effective inhibitor of rat renin in vivo. When infused (1 mg/kg/h) into two-kidney, one-clip chronic renal hypertensive rats, it lowered blood pressure and suppressed both plasma renin and angiotensin II. When given as a bolus (1 mg/kg) there was a divergence between the rapid rebound of renin levels and blood pressure, which remained suppressed. These results indicate that potent in vivo inhibitors of rat renin could be useful not only in examining the role of circulating renin but also in elucidating the equally important involvement of extracirculatory renin pools. PMID- 9352464 TI - Synthesis of phosphotyrosine-containing peptides using bis-(2,2,2-trichloro)ethyl groups for phosphate protection. AB - Suitability of bis-(2,2,2-trichloro)ethyl (Tc) groups for protection of phosphate moiety in Boc-mode synthesis of phosphotyrosine peptides is demonstrated Boc Tyr(PO3Tc2)-OH and Fmoc-Tyr(PO3Tc2)-OH were prepared by acylating H-Tyr(PO3Tc2) OH with (Boc)2O and Fmoc-ONSu, respectively. Phosphorus introduction was achieved by phosphorylating Boc-Tyr-OBzl with Tc phosphochloride. The Tc-phosphorus protector was found to be incompatible with the Fmoc group because the conditions of Fmoc removal (piperidine treatment) caused dephosphorylation. Complete NMR spectral assignments in the described compounds is presented. (Contribution No. 2398 from the Centre for Food and Animal Research). PMID- 9352463 TI - Synthesis and biological activity of potent, low molecular weight renin inhibitors. AB - A series of renin inhibitors containing the dipeptide transition state mimics (2R,4S,5S)-5-amino-4-hydroxy-2-methyl-6-cyclohexyl hexanoic acid (Cha psi[CH(OH)CH2]Ala) and (2R,4S,5S)-5-amino-4-hydroxy-2-isopropyl-6-cyclohexyl hexanoic acid (Cha-psi[CH(OH)CH2]Val) were prepared. A structure-activity study, using pseudopeptide (Boc-Phe-His-Leu-psi[CH(OH)CH2]Val-Ile-His-OH) as our lead structure, led to a new series of inhibitors, which correspond to tripeptides and contain no natural amino acids. For example, R,S-Bpma-Ape-Cha-psi[CH(OH)CH2]Ala NH2 (IC50 = 1.26 nM against human plasma renin at pH 6.0; molecular weight = 564) has only two thirds of the molecular weight but twice the potency of our original lead. This new class of low molecular weight renin inhibitor displays excellent specificity toward human renin versus the related aspartic proteinase pepsin and angiotensin-1-converting enzyme. Examples are given of selected inhibitors showing encouraging evidence for intestinal absorption after intracolonic and oral administration in male Sprague-Dawley rats. PMID- 9352465 TI - Synthesis and biological activities of fluorescent acridine-containing HIV-1 nucleocapsid proteins for investigation of nucleic acid-NCp7 interactions. AB - Specific interactions between the 72-amino acid nucleocapsid protein NCp7 of the human immunodeficiency virus, type 1 and the genomic RNA are essential for virus replication. Studies on the mechanism of action of NCp7 require a direct visualization of its complexes with nucleic acids and the determination of binding affinities. To facilitate these investigations, fluorescent NCp7 derivatives were developed by introduction in the NCp7 sequence of a non-natural amino acid, (S)-beta-(9-acridinyl)alanine (Aca) obtained by a chiral synthetic method. Three fluorescent NCp7 derivatives were obtained by introducing this amino acid at different positions. As shown by NMR, the three-dimensional structure of NCp7 is not altered by introduction of Aca. The fluorescent peptides were found to be as potent as their precursors in interacting with nucleic acids and in promoting HIV-1 genomic RNA dimerization. Moreover, because of their fluorescent properties, these NCp7s can be used at submicromolar concentrations to directly visualize and quantify protein-nucleic acid interactions in solution or after gel electrophoresis. This could facilitate the development of new antiviral agents aimed at inhibiting the functions of NCp7 and studies on the intracellular traffic of NCp7 within the preintegration complex. PMID- 9352466 TI - Structure-antitumor and hemolytic activity relationships of synthetic peptides derived from cecropin A-magainin 2 and cecropin A-melittin hybrid peptides. AB - The hybrid peptide (CA-ME) derived from cecropin A(1-8) and melittin (1-12) has potent antibacterial and antimalarial activities. Because the N-terminal sequence 1-12 of magainin 2 is similar to melittin(1-12), CA-MA with CA(1-8) and MA(1-12) and their analogues were designed and synthesized. Antitumor activities of these peptides were evaluated using three small cell lung cancer cell lines. Greater antitumor activity was observed when the residues 16, 18 and 19 of the peptide were hydrophobic (Leu or Val), basic (Lys) and basic (Lys), respectively. The IC50 values of the peptides with the residues were 2 to 4 microM. Residue 12 was related to hemolytic activity rather than antitumor activity. Increase in amphipathicity of P4 enhanced hemolytic activity without significant change in antitumor activity. The alpha-helicity of the peptides in a 30 mM sodium dodecyl sulfate solution was more closely correlated to hemolytic activity than antitumor activity. PMID- 9352467 TI - Conformational analysis of potent sweet taste ligands by nuclear magnetic resonance, computer simulations and X-ray diffraction studies. AB - Four potent sweet-tasting molecules, N-(3,3-dimethylbutyl)-L-aspartyl-L phenylalanine methylester 1 (7000 times more potent than sucrose), N-(3,3 dimethylbutyl)-L-aspartyl-D-valine (S)-alpha-ethylbenzylamide 2 (3000 time more potent than sucrose), L-aspartyl-D-valine (R)-alpha-methoxymethylbenzylamide 3 (1350 times more potent than sucrose and L-aspartyl-(1R,2S,4S)-1-methyl-2-hydroxy 4-phenylhexylamide 4 (2500 times more potent than sucrose) were studied by 1H NMR and computer simulations. These flexible molecules adopt multiple conformations in solution. The "L-shaped" structure, which we believe to be responsible for sweet taste is accessible to all four compounds in solution. Extended conformations with the AH and B-containing moieties in the +y-axis and the hydrophobic group X pointing in the y-axis have also been observed for all four sweeteners. For compounds 1 and 3, the solid-state conformations were determined by X-ray diffraction studies. These results demonstrate that compounds 1 and 3 adopt an "L-shaped" structure even in the crystalline state. The extraordinary potency of the N-alkylated compound 1 compared with the unsubstituted Asp-Phe-OMe may be explained by the effect of a second hydrophobic binding domain in addition to interactions arising from the "L-shaped" structure. PMID- 9352469 TI - Granulocyte-colony stimulating factor maintains a thermally stable, compact, partially folded structure at pH2. AB - At acidic pH many proteins exist in a partially unfolded form, called the "A" state. This is defined as a flexible, expanded structure with well-defined, usually native-like secondary structure, but no unique tertiary structure, and showing no cooperativity during thermal-induced denaturation. Granulocyte-colony stimulating factor (G-CSF), a four-helix bundle cytokine, maintains both thermal stability and tertiary structure at pH 2.0. We therefore examined the conformation and thermal unfolding of G-CSF at pH 2.0, 4.0 and 7.0 using circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR). The secondary structure of the molecule remains highly helical as the pH is lowered from 7.0 to 2.0. The tertiary structure of the protein is slightly different at each pH value, but even at pH 2.0 G-CSF maintains a regular three-dimensional structure. The structure is hydrodynamically compact at these different pH values, with no increase in Stoke's radius even at pH 2.0. The thermal-induced denaturation of G CSF was determined by monitoring changes in the CD or FTIR spectra. At pH 2.0 the temperature at which thermal-induced denaturation begins is higher than it is at pH 4.0 or 7.0, the thermal unfolding transition remains cooperative and some alpha-helical structure persists even at 86 degrees C. At pH 4.0 and 7.0, secondary and tertiary structures disappear simultaneously during thermal denaturation, whereas at pH 2.0 small changes in the far-UV CD region begin to occur first, followed by the simultaneous cooperative loss of tertiary structure and much of the remaining secondary structure. The structure of G-CSF at pH 2.0 is thus revealed as compact, with a unique, three-dimensional structure, highly helical secondary structure, and most importantly, a cooperative thermal unfolding transition. G-CSF at acid pH thus does not adopt the "A" state. PMID- 9352468 TI - Comparison of solution properties of human and rat ciliary neurotrophic factor. AB - The solution structure and stability of rat and human ciliary neurotrophic factor (CNTF) were examined by circular dichroism (CD), Fourier transform infrared (FTIR) and fluorescence spectroscopy and sedimentation equilibrium analyses. The secondary structure of both proteins, as assessed by CD and FTIR, consists primarily of alpha-helix, consistent with CNTF being a member of the four-helical bundle family of cytokines and neurokines, with rat CNTF containing slightly less helix (about 10% less) and slightly more disordered structure. The environment of the tyrosine and tryptophan residues, assessed by intrinsic fluorescence emission spectroscopy, appears to be the same in both proteins. Binding of anilinonaphthalene sulfonate is identical for both proteins, indicating that these two proteins have similar surface hydrophobicities in the native state. The thermal stability of the human CNTF is significantly less than that of the rat CNTF, yet their stabilities to guanidine HCl-induced denaturation are equivalent. This apparent discrepancy in stability between the two proteins may be explained by solubility differences upon thermal unfolding. Although the human protein precipitates as it is denatured by heat, the rat protein does not. It thus appears that the unfolded state of human CNTF is less soluble and more prone to aggregation than that of the rat protein upon heating, although their conformational stability is similar. Both proteins remain largely folded at pH 3.0. Sedimentation equilibrium analysis demonstrates that both rat and human CNTF exist primarily as monomers; however, significant dimer formation occurs as the protein concentrations are increased to greater than 3 mg/mL, particularly in the presence of ammonium sulfate. PMID- 9352470 TI - The functional neuroanatomy of mood disorders. AB - Mood disorders may be associated with global and regional changes in cerebral blood flow and metabolism. The accumulated functional neuroimaging findings in mood disorders were reviewed in order to examine a proposed neuroanatomic model of pathophysiology. Global cerebral blood flow and glucose metabolism appear normal, but may be decreased in late-life depression. Regional cerebral blood flow and glucose metabolism deficits are present, and may be indicators of brain regions participating in neuroanatomic circuits involved in mood disorders. Decreased pre-frontal cortex blood flow and metabolism in depressed unipolar and bipolar patients are the most consistently replicated findings, and correlate with severity of illness. Basal ganglia abnormalities have been found in depressed unipolar and bipolar patients, involving decreased blood flow and metabolism. Temporal lobe abnormalities are present in bipolar disorder patients, and perhaps unipolar depression. There is conflicting evidence of abnormalities in other limbic regions. Cognitive impairment may correlate with decreased metabolism in frontal and cerebellar areas. The relationship between functional neuroimaging findings and clinical course, and therefore state and trait characteristics, has not been systematically investigated. Antidepressant medications, but not ECT, seem to reverse some of the identified functional brain changes in the depressed state. The structural, neurotransmitter and neuropathological correlates of these functional abnormalities are yet to be determined. Functional abnormalities in frontal, subcortical and limbic structures appear to be part of the pathophysiology of mood disorders. PMID- 9352471 TI - Critical analysis of the theories advanced to explain short REM sleep latencies and other sleep anomalies in several psychiatric conditions. AB - One of the most consistent and most studied sleep modifications in several psychiatric conditions is the shortening of the rapid eye movement (REM) sleep latency. While its clinical usefulness is still to be proven and its meaning relatively obscure, the appearance of a short REM latency continues to be a daily fact in sleep laboratories. Many theories compete to explain what is observed, the most important being the circadian rhythm hypotheses, the homeostatic model and the reciprocal interaction model. These three are summarised and their pros and cons are exposed in a systematic manner. Points of conflict, possible convergences and limitations are discussed in the light of recent developments on the general theories of sleep regulation. PMID- 9352472 TI - Impulsivity in self-mutilative behavior: psychometric and biological findings. AB - This paper examines impulsivity as a central factor in moderate/superficial self mutilation such as skin-cutting and burning. A sample of 165 subjects were divided into four groups, namely self-mutilators, patients with any modes of impulsive behavior other than self-mutilation, patients without any impulsive behavior, and normal probands. All were administered the 10th version of the Barratt Impulsiveness Scale, the State-Trait Anger Expression Inventory, and the Inventory for the Assessment of Factors of Aggressiveness. They also were interviewed carefully in regards to both impulsive and self-mutilative behavior. A d-fenfluramine challenge test was administered to 36 females and prolactin levels were measured. On the whole results implicate impulsive personality functioning as a major factor in subjects with moderate/superficial self mutilative behavior whose trait pathology is similar to personality disordered patients with other modes of self-harming impulsive behavior. PMID- 9352473 TI - Personality factors and weight preoccupation: a continuum approach to the association between eating disorders and personality disorders. AB - OBJECTIVE: Evidence shows a high comorbidity of eating disorders and some forms of personality disorder. Adopting a dimensional approach to both, our study explored their connection among a non-clinical sample. METHOD: 191 young women completed personality scales of general neuroticism, and of borderline, schizotypal, obsessive-compulsive, and narcissistic (both adjustive and maladaptive) traits. Weight preoccupation (WP), as a normal analogue of eating disorders, was assessed with scales from the Eating Disorder Inventory, and height and weight measured. The data were analysed with multiple regression techniques, with WP as the dependent variable. RESULTS: In low to normal weight subjects, after controlling for the significant influence of body mass, the specific predictors of WP in the regression model were borderline personality and maladaptive narcissism, in the positive direction, and adjustive narcissism and obsessive-compulsiveness in the negative direction. In heavier women, narcissism made no contribution--nor, more significantly, did body mass. CONCLUSIONS: Patterns of association between eating pathology and personality disorder, especially borderline and narcissism, can be clearly mapped across to personality traits in the currently non-clinical population. This finding has important implications for understanding dynamics of, and identifying individuals at risk for, eating disorders. PMID- 9352474 TI - Panic-associated suicidal and aggressive ideation and behavior. AB - Some investigators have noted an increased incidence of suicidal ideation and attempts in individuals with panic attacks. The direct temporal relationship between the panic state and suicidal thoughts and behaviors has not been well elucidated however. Furthermore, although aggressive behavior is often manifested in individuals with suicidal behavior, the relationship between aggression and panic has received little attention. The aim of this study was to assess the frequency and type of reported suicidal and aggressive ideation and behaviors that occur during the panic state in patients with panic disorder. In order to evaluate the contribution of depression, individuals with pure (i.e. uncomplicated) panic disorder were compared with individuals who had comorbid panic and major depression. Nineteen patients with a diagnosis of pure panic disorder and 28 patients with comorbid panic plus major depression were included in the study. All patients were given the Panic, Suicide and Aggression Scale (PSAS), a questionnaire specifically designed to assess reported suicidal and aggressive thoughts and behaviors that occur during panic attacks. Other scales given to all patients included overall measures of impulsivity, suicide risk and violence risk. Patients with pure panic disorder reported high rates of suicidal and aggressive ideation and behavior during panic. The presence of comorbid depression resulted in a doubling of the rate of reported panic-associated suicidal ideation, property destruction and assaults, and a five-fold increase in the rate of homicidal ideation. The rate of reported suicide attempts was equal in the pure panic and comorbid group. There were also high correlations in all panic patients between measures of panic-associated suicide and aggression with the psychometric measures of impulsivity, suicide risk and violence risk. PMID- 9352476 TI - LASIK for myopia and astigmatism after penetrating keratoplasty. PMID- 9352475 TI - Acute inositol does not attenuate m-CPP-induced anxiety, mydriasis and endocrine effects in panic disorder. AB - Many anti-panic drugs, administered chronically, can block pharmacologically induced "panic attacks"; acutely they often exacerbate panic disorder. Theories of action need to account for this biphasic effect. Chronic inositol had previously shown efficacy against panic disorder. The authors investigated the effect of a single dose of 20 g inositol on an m-CPP challenge in a double-blind placebo-controlled crossover trial in panic-disorder patients. Seven patients had robust psychological, physiological and endocrine responses to 0.08 mg m-CPP i.v.; inositol had virtually no effect on these responses, although it had some acute effects during the evening before the challenge. A similar trial involving chronic inositol would be of interest. PMID- 9352477 TI - Intraocular use of hyaluronidase to dissolve sodium hyaluronic acid. PMID- 9352479 TI - Excimer laser in situ keratomileusis to correct compound myopic astigmatism. AB - PURPOSE: We studied the efficacy, predictability, stability, and safety of excimer laser in situ keratomileusis (LASIK) to correct myopia and astigmatism. METHODS: We prospectively studied 87 consecutive eyes of 56 patients who received LASIK, divided into two groups: the myopic group included eyes with myopia more than -2.00 diopters (D) and astigmatism less than 0.50 D and the astigmatism group included eyes with myopia of more than -2.00 D and astigmatism of 0.50 D or more. The Chiron automated corneal shaper and the Nidek EC-5000 excimer laser were used in all eyes. A modified personal nomogram was used in all eyes. The changes in refractive sphere and cylinder, and complications were studied at 2 and 6 weeks, 3, 6, and 12 months after surgery. Preoperatively, the mean spherical equivalent refraction was -4.41 D (range, -2.25 to -7.25; SD, 1.74) in the myopia group and -5.79 D (range, -2.25 to -15.50 D; SD, 2.45) in the astigmatism group. The mean spherical component of the refraction was -4.39 D (range, -2.25 to -7.25; SD, 1.74) in the myopia group and -5.19 D (range, -2.00 to -14.00; SD, 2.32) in the astigmatism group. The mean refractive cylinder was 1.19 D (range, 0.5 to 3.00 D; SD, 0.62) in the astigmatism group. RESULTS: At 12 months, 81 eyes (93.6%) of 51 patients were examined; the mean spherical equivalent refraction was -0.43 D (range, +0.50 to -1.25 D; SD, 0.35) in the myopia group and -0.33 D (range, +1.25 to -2.13 D; SD, 0.52) in the astigmatism group. The mean spherical component of the refraction at 12 months was -0.33 D (range, +0.50 to -1.25 D; SD, 0.33) in the myopia group and -0.17 D (range, +1.50 to -1.50; SD, 0.48) in the astigmatism group. The mean refractive cylinder was 0.19 D (range, 0 to 0.75 D; SD, 0.25) in the myopia group and 0.32 D (range, 0 to 1.25 D; SD, 0.30) in the astigmatism group. The mean change in spherical equivalent refraction between 6 weeks and 12 months after surgery was -0.08 D toward myopia (range, -0.50 to -0.75 D; SD, 0.23) in both groups. No eyes lost two or more lines of spectacle-corrected visual acuity. Patient satisfaction was high in both groups. Complications included undercorrection that necessitated reoperation (three eyes), overcorrection (two eyes), and small disc diameter (one eye). No vision threatening complications were observed. CONCLUSION: LASIK with the Nidek EC5000 laser is effective, reasonably predictable, stable, and safe for correction of compound myopic astigmatism with a spherical component between 2.00 and -14.00 D, and a cylindrical component between 0.50 and 3.00 D using the techniques in this study. Astigmatism is undercorrected with the current algorithm. Correction of higher amounts of astigmatism requires further study. PMID- 9352478 TI - Excimer laser photorefractive keratectomy for hyperopia. AB - OBJECTIVE: Excimer laser photorefractive keratectomy (PRK) has been shown to be an effective method in the treatment of refractive errors, especially myopia. We evaluated prospectively the efficacy, predictability, stability, and safety of excimer laser PRK in the treatment of hyperopia. METHODS: Thirty-four hyperopic eyes were treated with an Aesculap-Meditec (MEL 60) excimer laser. The patients were divided into two groups. In the low-moderate hyperopia group, baseline spherical equivalent refraction was between +1.50 and +6.00 diopters (D) (mean, +4.20 +/- 1.30 D) and in the high hyperopia group between +6.25 and +9.75 D (mean, +7.70 +/- 1.30 D). Follow-up visits occurred 1, 3, 6, and 12 months after surgery. RESULTS: One-year results were available for a total 27 eyes (79%): 15 eyes with low to moderate hyperopia and 12 eyes with high hyperopia. One year after PRK in the low-moderate group, six eyes (40%) had a refractive error within +/- 1.00 D of emmetropia, but in the high hyperopia group only two eyes (17%) were within +/- 1.00 D of emmetropia; three eyes (20%) and one eye (8%) were within +/- 0.50 D, respectively. The stability of the refractive change was better in the low to moderate hyperopia group; in the high hyperopia group there was still some regression after 6 months. At 12 months, 10 eyes (67%) in the low moderate and one eye (8%) in the high hyperopia group had postoperative uncorrected visual acuity of 20/40 or better. One eye in the low-moderate hyperopia group saw 20/20 without correction. Only one eye lost two lines of spectacle-corrected visual acuity. Haze was more intense in the high hyperopia group, but it did not reduce visual acuity. No vision-threatening complications were observed. CONCLUSIONS: When low to moderate hyperopia up to +6.00 D is treated, excimer laser PRK with the Aesculap Meditec MEL60 laser is safe and moderately effective, and refraction stabilizes after 3 months in most eyes. However, PRK is not sufficient to treat high hyperopia in an effective and predictable way. PMID- 9352480 TI - Excimer laser in situ keratomileusis for myopia. AB - PURPOSE: To evaluate the efficacy, safety, and predictability of excimer laser in situ keratomileusis (LASIK) for the correction of myopia. METHODS: Forty-six consecutive eyes that had LASIK with the VISX 20/20B laser and Chiron corneal shaper were evaluated. Mean spherical equivalent of the preoperative manifest refraction was -9.40 +/- 3.78 diopters (D) (range, -3.50 to -19.75 D). The refractive effect (postoperative refraction minus baseline refraction), residual refractive error (refractive effect minus planned correction) and uncorrected and spectacle-corrected visual acuity were examined. RESULTS: Mean follow-up was 6.1 months (range, 3 to 9 mo). Spectacle-corrected visual acuity was unchanged in 39 eyes (84.78%), significantly improved in five eyes (10.86%), and worse in two eyes (4.34%). Uncorrected visual acuity was 20/20 or better in 15 eyes (32.6%) and 20/40 or better in 39 eyes (84.78%). Thirteen eyes (28.26%) had a postoperative spherical equivalent refraction within +/- 0.50 D and 36 eyes (78.26%) within +/- 1.00 D of attempted correction. No intraoperative complication occurred. Postoperative complications were few and not severe: three eyes (6.52%) developed regular astigmatism, two eyes (4.34%) had interface deposits, and three eyes (6.52%) had small epithelial cysts in the interface. CONCLUSION: LASIK with the VISX 20/20B laser is safe, moderately effective, and relatively predictable for correcting myopia from -3.50 to -19.50 D. Predictability decreases with increasing myopia. PMID- 9352481 TI - Comparison of laser in situ keratomileusis and photorefractive keratectomy to correct myopia from -1.25 to -6.00 diopters. AB - BACKGROUND: We evaluated the safety and efficacy of laser in situ keratomileusis (LASIK) for the correction of low to moderate amounts of myopia (-1.25 to -6.00 D). METHODS: Photorefractive keratectomy (PRK) was performed on 432 eyes and LASIK on 137 eyes with a Chiron Keracor 116 excimer laser. Uncorrected and corrected visual acuity, corneal sensitivity, contrast sensitivity, and corneal topography were examined before and after surgery. RESULTS: One-year follow-up of 307 eyes in the PRK group and 103 eyes in the LASIK group was achieved. At 1 year, 83% (85 of 103) of LASIK eyes and 72% (221 of 307) of PRK eyes had an uncorrected visual acuity of 1.0 or better. Eighty-nine percent (92 of 103) of LASIK eyes and 83% (255 of 307) of PRK eyes had a refractive error within +/- 1.00 D of emmetropia; 71% (73 of 103) of LASIK eyes and 61% (188 of 307) of PRK eyes were within +/- 0.50 D of emmetropia. Contrast sensitivity and corneal sensitivity were reduced in both groups at the early postoperative stage but gradually returned to preoperative values; their recovery took about 3 months in LASIK eyes and 6 to 12 months in PRK eyes. CONCLUSION: LASIK is safe and more predictable than PRK to correct low to moderate amounts of myopia. Recovery from LASIK is faster than after PRK. PMID- 9352482 TI - Three multizone photorefractive keratectomy algorithms for myopia. The Melbourne Excimer Laser Group. AB - OBJECTIVE: To compare the efficacy and complications of three different excimer laser algorithms for multizone photorefractive and photoastigmatic keratectomy. METHODS: Three different software algorithms were applied to treat myopia and myopic astigmatism with the VISX 20/20 excimer laser. Each algorithm had a maximum ablation zone of 6 mm but differed in the number of zones employed, the proportion of the total treatment allocated to each ablation zone, and the treatment of astigmatism. The Melbourne multizone technique equally divided myopia correction into a maximum of three ablation zones. The Pop multizone technique biased myopia treatment into the smaller diameter zones to a maximum of six ablation zones, with one central island pretreatment. The Alpins multizone technique equally divided myopia treatment through all zones up to a maximum of six, with one central island pretreatment. RESULTS: A total of 585 patients (780 eyes) were treated and 625 eyes (80%) were followed for more than 6 months. The mean baseline spherical equivalent refractive error was -5.63 D (-1.00 to -19.50 D). Between 71 and 79% of eyes were treated for astigmatism. There was no statistically significant differences in baseline refractive error or other characteristics among the three groups. At 6 months, the Alpins multizone algorithm had more eyes with a refractive error within +/- 1.00 D of emmetropia (p = 0.01) and more within +/- 2.00 D of emmetropia (p < 0.01). This new algorithm produced more eyes with an uncorrected visual acuity of 20/20 or better at 6 months (p < 0.01). When multiple logistic regression was used to correct for any differences in baseline myopia among the three groups, this algorithm also had a higher odds ratio for achieving 20/20 or better uncorrected visual acuity (OR = 1.58). CONCLUSION: At 6 months, all three algorithms were effective in the reduction of myopia. Significantly better visual acuity and refractive results were achieved with the Alpins multizone algorithm that spread the total treatment over a larger number of ablation zones, with an equal number of diopters of treatment in each zone. PMID- 9352483 TI - Iris claw phakic intraocular lens for high myopia. AB - BACKGROUND: The implantation of a Worst-Fechner iris claw intraocular lens (IOL) is one of the surgical procedures used for the correction of high myopia. This technique reduces myopia with stable refractive results; however, its potential long-term risks have not been evaluated. We report results in 94 eyes with a minimum follow-up of 3 years. METHODS: We studied 94 eyes of 62 patients with myopia > or = -7.00 diopters (D) who underwent Worst-Fechner IOL implantation. Lens decentration, permeability of the blood-aqueous barrier by iris angiography, and changes in corneal endothelial density were analyzed. RESULTS: Mean follow-up time was 48.9 months (range 36 to 72 mo). Three years after surgery, 58 eyes (61%) had an uncorrected visual acuity > or = 20/40, and 77 eyes (82%) gained two or more lines of spectacle-corrected visual acuity with respect to the preoperative value; 75 eyes (79%) were within +/- 1.00 D of emmetropia and 46 eyes (48%) were within +/- 0.50 D of emmetropia. The mean endothelial cell loss was 17.9% at 5 years after surgery, while the percentage of hexagonality and the coefficient of cell variation tended toward preoperative levels. No vision threatening complications were seen. CONCLUSIONS: The implantation of a Worst Fechner iris claw phakic IOL reduced high myopia with a stable refractive outcome. Endothelial cell damage was within acceptable limits. The absence of major complications makes this procedure an acceptable method for correcting high myopia. PMID- 9352484 TI - Corneal asphericity in eye bank eyes implanted with the intrastromal corneal ring. AB - OBJECTIVE: To evaluate the effects of the intrastromal corneal ring, a device developed to reduce myopia, on corneal asphericity in a large set of eye bank eyes. METHODS: Forty-one deturgesced eye bank eyes were implanted with intrastromal corneal rings of five different thicknesses, ranging from 0.25 mm to 0.45 mm. Corneal asphericity, before and after implantation, was examined using two different metrologies. Corneal asphericity profiles were produced from dioptric power data collected from videokeratography. To statistically assess the corneal asphericity differences between exam times for each intrastromal corneal ring thickness, dependent sample confidence intervals (95%) were calculated for the mean differences between preoperative and postoperative measures for each topographic diameter zone. Laser holographic interferometry was used to inspect corneal asphericity in one eye bank eye case study for four intrastromal corneal ring sizes. Wave unit map and geometric zonal spot ray tracing analyses derived from laser holographic interferometry topography were surveyed. RESULTS: Videokeratographic analysis suggested that preoperative corneal shape was prolate, i.e., flattened from central to paracentral cornea. Corneal shape became more prolate with intrastromal corneal ring implantation for all intrastromal corneal ring thicknesses. Laser holographic interferometry demonstrated that prolate asphericity was preserved with the intrastromal corneal ring sizes tested and that optical collection efficiency of the cornea was not diminished. CONCLUSION: Using two different measurement techniques, this eye bank eye study demonstrated that intrastromal corneal rings maintain prolate corneal asphericity. PMID- 9352485 TI - Dichlorotriazinyl aminofluorescein inhibits human keratocyte proliferation in vitro. AB - PURPOSE: Dichlorotriazinyl aminofluorescein (DTAF) has been used to stain corneal stromal collagen as part of in vivo experimentation. Toxicity of this drug, if present, might alter the observed wound healing. To determine if this drug has any deleterious effect on keratocytes, we evaluated it in vitro. METHODS: Human keratocytes in 96 well plates were exposed to different concentrations of DTAF (10e-7, 10e-6, 10e-5, 10e-4, 10e-3, 10e-2, and 10e-1 mg/ml of media). Exposure times of 1 and 24 hours at each concentration of DTAF were evaluated. The cell number was measured 1 and 3 days after exposure to the drug using a coulter counter and a hemocytometer. RESULTS: The proliferation of keratocytes after 24 hours of exposure to the drug was inhibited in a dose dependent manner by DTAF, but 1 hour exposure of keratocytes to the drug did not inhibit keratocyte proliferation. CONCLUSION: These results suggest that DTAF has inhibitory effects on human keratocyte proliferation after 24 hours of exposure, while exposure limited to 1 hour does not induce such a change. PMID- 9352486 TI - A standardized classification of corneal topography after laser refractive surgery. PMID- 9352487 TI - Checklist of information usually submitted in an investigational device exemption (IDE) application for refractive surgery lasers. Ophthalmic Devices Advisory Panel, Food and Drug Administration. PMID- 9352488 TI - Removal of age-related cataract and iris claw phakic intraocular lens. PMID- 9352489 TI - Transgenic mouse models for studying mutations in vivo: applications in aging research. AB - To study mutation accumulation in the DNA of somatic cells and tissues during aging in vivo, a transgenic mouse model has been constructed. The model harbors plasmid vectors, containing the lacZ reporter gene, integrated head to tail at various chromosomal locations. Procedures have been worked out to efficiently recover the plasmids into E. coli host cells. A positive selection system, permitting only E. coli cells with a lacZ mutated plasmid to grow, allows for the accurate determination of mutation frequencies as the ratio of mutant colonies versus the total number of transformants, i.e., the total number of plasmid copies recovered. Results obtained from a life span study of plasmid mice with vector clusters on chromosome 3 and 4 indicated age-related mutation accumulation in the liver, but not in the brain. Comparison of the mutational spectra revealed a significantly larger proportion of large size-change mutations in liver than in brain. PMID- 9352491 TI - Molecular genetic approaches to the genes of longevity, aging and neurodegeneration in mammals. AB - Accumulative evidence suggests that species life-span is determined, at least in part, genetically. Recent cloning works using yeast (Saccharomyces cerevisiae) and worms (Caenorhabditis elegans) revealed several potential candidate genes, e.g. SIR4, a transcriptional silencing factor, and age-1, a putative signal transduction molecule, respectively, that may be involved in determining and/or regulating species life-span in lower organisms. It is, however, not clear yet whether mammalian homologs of these genes are also relevant to controlling longevity in higher organisms. In mice and humans several silencing factors are essential for cell-type specific gene expression. A variety of signal transducing molecules are also known to play important roles in mammals. I will briefly summarize recent progress in molecular genetic studies on such longevity-related genes, and discuss these results with our recent findings on a neural-selective silencing factor and a neural-specific signaling molecule that are important for functioning of the nervous system. PMID- 9352490 TI - Analysis and modulation of DNA repair in aging. AB - Nearly 40 years ago it was proposed that accumulation of mutations or increased levels of DNA damage might contribute to aging processes. Despite several correlative studies in this area, the answer as to whether genomic integrity contributes to aging has remained illusive. More recently it has been hypothesized that decreased mitochondrial DNA integrity plays a role in aging. To begin to test these hypotheses more directly, we are developing transgenic mouse and cell culture model systems. For example, transgenic mice overexpressing the DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT) have been made and have a reduced spontaneous frequency of hepatocellular carcinoma. A lifespan study using the MGMT transgenic mice is in progress in an effort to determine whether cancer impacts on the median or maximal lifespan of a species. Second, a quantitative PCR technique is being used to measure mitochondrial DNA damage in mitotic and post-mitotic cells to determine if the level of damage and/or repair is different based on mitotic status. Finally, mice deficient in metallothionein I and -II are being used in an effort to determine if the subcellular distribution of metals impact on oxidative damage with increased age. PMID- 9352492 TI - Age changes in signal transduction and gene expression. AB - Altered regulation of physiological and behavioral processes is an important functional manifestation of aging. Our laboratory has been examining a number of model systems in order to elucidate the mechanisms by which these processes, controlled mainly by hormones and neurotransmitters, change during aging. Two, in which alternations in gene expressions are critical, are loss of striatal dopaminergic motor control and impaired stimulation of hepatocyte DNA synthesis. Loss of striatal D2 dopamine receptors contributes substantially to reduced motor control in the elderly. Such receptor loss is due both to the death of some receptor-containing neurons and decreased expression of the receptor gene in the surviving neurons. Current efforts are focussed on the mechanisms responsible for neuronal death, reduced gene expression and the relationship between the two. In addition, the D2 receptor gene has now been inserted into attenuated adenoviral vectors which elicit expression of functional receptors when injected into the brains of living rats and mice. Stimulation of DNA synthesis by various agents including catecholamines and growth factors is markedly reduced in primary cultures of hepatocytes obtained from aged rats when compared with younger counterparts. Such impairment is not the consequence of receptor loss. Moreover, since very different signal transduction pathways are employed by G protein linked receptors and those mediated by tyrosine kinases, the defect would appear to be at a very functional level. Results to date indicate that increased expression of sdi-1/p21, an inhibitor of cyclin-dependent kinases, is not responsible. However, decreased stimulation of the MAP Kinase pathway, possibly due to elevated levels of MAP Kinase Phosphatase, may also play a role. IN addition, cells of aged rats appear to shift to other growth factor responsive pathways. In summary, altered gene expression during aging may be responsible for some important impairments in signal transduction and corresponding physiological and behavior functions. PMID- 9352493 TI - Hierarchical deterioration of body systems in Werner's syndrome: implications for normal ageing. AB - Normal human ageing is a complicated biological phenomenon. 'Werner's syndrome (WS)', caused by mutations of RecQ type DNA helicase, has been recognized as a top ranking 'segmental' progeroid syndrome. Patients with WS show a wide variety of clinical and biological manifestations in the four major self-assembly body systems (nervous, immune, connective tissue and endocrine-metabolic systems) similar to normal ageing at an early stage of their life, followed by death at an average age of 46. The sequential appearance of clinical and biological deterioration of the body systems observed in WS suggested that the disorder is more than a segmental progeroid syndrome, analysis of which may shed new light on the question 'Why and how we age?' PMID- 9352494 TI - Molecular and epidemiological studies of Werner syndrome in the Japanese population. AB - Werner syndrome (WS) is an autosomal recessive genetic disease characterized by many age-related features. The gene responsible for WS (WRN) has been isolated and contains a helicase domain, but its function is unknown. Six different mutations throughout the WRN gene have been reported in the Japanese population. We have studied whether patients with a specific mutation exhibit distinct phenotypes from others. Fourteen patients with different mutations showed almost the same signs and symptoms and, therefore, the C terminal part of the product appears to be crucial for its functions, although other parts may be important as well. Haplotype analyses using 13 microsatellites covering the 2.8-3.0 cM WRN region showed that two out of six different mutations had founder chromosomes. These two founder chromosomes may be evenly distributed throughout the western part of Japan, suggesting that these mutations go back to a time earlier than 1400 years ago. PMID- 9352495 TI - Coronary artery disease associated with Helicobacter pylori infection is at least partially due to inadequate folate status. AB - The numerous effects of Helicobacter pylori have attracted significant attention. The most consistent and well appreciated effect is peptic ulcer. However, gastric cancer, growth retardation and coronary artery disease are among other sequelae of this chronic infection. This discussion describes a potential relationship among risk of coronary artery disease, the changes caused in gastric juice by H. pylori-induced gastritis, and the bioavailability of folates. Reduced folate absorption can occur in an environment of increased gastric juice pH and/or decreased ascorbic acid. This can, relatively rapidly, result in inadequate folate status which inhibits the methionine synthase reaction. Reduced methionine synthase activity increases the blood concentration of homocyst(e)ine which is known to be toxic to endothelial cells, and an independent risk factor for atherosclerosis. Decreased folate bioavailability may help explain the increased risk of coronary artery disease which has been observed in populations infected with H. pylori. It would also be consistent with the increased occurrence of this association in lower socioeconomic groups, and may also help explain the low incidence of gastric cancer in Africa, despite the high prevalence of H. pylori infection. PMID- 9352497 TI - Targeting a key enzyme in cell growth: a novel therapy for cancer. AB - The enzyme ribonucleotide reductase (RR) controls the synthesis of DNA precursors and thus plays a pivotal role in cell growth. Since the free-radical-containing active-site of this enzyme can be disabled by a lone electron, low-level direct electric current should have an inhibitory effect on RR and, thus, on uncontrolled cell proliferation. This hypothesis is strongly supported by the results of several cancer electrotherapy studies reported over the years. PMID- 9352496 TI - Abdominal hypertension and disproportion: a universal and fundamental disorder which varies from minor to major but is not well known. AB - Abdominal hypertension/disproportion occurs in older patients as hernias (inguinal, umbilical, diaphragmatic, recurrent, etc.), in immature mothers whose uteri and abdomens are not ready for pregnancy, and as postoperative distention and abdominal wound dehiscence. PMID- 9352498 TI - Parapsychotic grief, theory of mind and the concept of the soul. AB - The ability to deceive is regarded as the best evidence of the cognitive ability separating humans from other primates. An alternative would be to look at the concept of the soul, which has an archetypal significance, emerging in various geographically remote cultures over the course of history. The soul will be an elusive but not an impossible concept to study with neuroimaging. In parapsychotic grief the decreased may appear to the bereaved person without these hallucinations being considered as indicative of mental illness. If this is the sort of normal human experience which has led to the emergence of the belief in the immortality of the soul it may be a useful starting point for defining the neuroanatomical basis of souls which do not necessarily seek to deceive. As the human prefrontal cortex expanded and developed and strove to understand mental activity derived from subcortical structures the human attained an awareness of his own mind which has been construed as a separable insubstantial but indestructible entity. This idea would be bizarre if it were not archetypal and therefore must be closely linked to the development of the human central nervous system. PMID- 9352499 TI - Could nitroglycerine poisoning be the cause of Alfred Nobel's anginal pains and premature death? AB - The life of 19th century Swedish chemist cum inventor Alfred Nobel can be conveniently divided into two equal phases: pre-nitroglycerine phase (1833-1863) and nitroglycerine phase (1864-1896). According to the records of Ragnar Sohlman, his assistant during his last year of life, Nobel's physical condition began to decline towards the end of the 1870s, and for the last 16 years he suffered from deep depression and anginal pains. Based on Nobel's descriptions of his condition, on his prolonged experimentation with explosives, his strenuous work habit and some recent knowledge about nitroglycerine poisoning, I hypothesize that nitroglycerine poisoning was an aggravating factor which contributed to Nobel's deteriorating health and premature death at the age of 63. PMID- 9352500 TI - Multiple sclerosis: an immune legacy? AB - The aetiology of multiple sclerosis suggests that its occurrence depends on a combination of factors, including viral infection in early childhood, genotype and an initiating event within the central nervous system. The resulting damage may be caused by events initiated by free radicals. Free radicals themselves may cause damage to myelin and also may trigger the arachidonic acid cascade, to produce compounds that are thought to initiate and augment T-cell activity. Repair of the damaged tissue is normally achieved by protective enzymes that remove damaged lipid from the myelin. PMID- 9352501 TI - The French paradox unmasked: the role of folate. AB - The French paradox relates to the paradoxical association of a diet high in saturated fat and cholesterol with low coronary heart disease mortality and is contrary to the 'lipid hypothesis'. France and other regions with low heart disease mortality have a high consumption of fruit and vegetables. Epidemiologic studies show fruit and vegetable consumption is inversely related to coronary heart disease mortality, but recent intervention studies do not support the theory that protection is due to antioxidant vitamins. Fruit and vegetables, however, are rich sources of folate. Folate lowers plasma homocysteine levels. Even mild to moderate elevation in plasma homocysteine level is a strong risk factor for arteriosclerosis of the coronary, cerebral, and peripheral arteries. This should explain not only the French paradox but also why known risk factors may explain as little as 25% of the risk for coronary heart disease. PMID- 9352502 TI - A proposed new strategy of immunotherapy for Alzheimer's disease. AB - Current data on the involvement of the immunological system in the pathogenesis of Alzheimer's disease (AD) are discussed, and results of immunotherapy for the disease are provided. Hypotheses on immune aging as a risk factor for AD, and a suggested new treatment strategy, are presented and discussed. PMID- 9352503 TI - A theoretical analysis of oriental medicine, 1: Acupuncture. AB - In spite of its long history, oriental medicine has been little understood, and therefore poorly accepted, by the Western mind. In order to penetrate beyond the veil of its jargon, a theoretical analysis of the fundamental tenets of acupuncture is given here, using concepts and symbols of modern science to elucidate phenomena of everyday practice. Thus, when Chi, Yang, and Ying are regarded as 'phased flows of bio-electromagnetic energy', intuitive comprehension should be facilitated. This novel approach has not, to the author's knowledge, been applied previously to any aspect of the medicine of the 'mysterious East'. A similar approach will be used to explore Kampo diagnosis/therapeutics and other selected topics in later papers. PMID- 9352504 TI - Parkinson's disease, amyotrophic lateral sclerosis and spinal muscular atrophy are caused by an unstable (CAG)n trinucleotide repeat microsatellite. PMID- 9352506 TI - Diseases of plants transmissible between plants and man (phytonoses) exist. AB - Since zoonoses (singular zoonosis) are infections or infectious diseases of animals transmissible to man, or occasionally from man to other animal species, it is logical that should any diseases of plants be found that are transmissible between plants and man they should be termed phytonoses (singular phytonosis). The genome of human uveitis mycoplasma-like organisms (MLO) has been reported to be showing a very high homology to plant MLO. For this reason, and because there are several other known similarities between plant and human MLO, it is hypothesized that at least one class of phytonoses, namely cross-infection of MLO between plants and man, exists. There are, however, other possibilities. Cross infection transmission experiments, the results of which could add credibility to the hypothesis, could be undertaken. PMID- 9352505 TI - Zinc-induced suppression of inflammation in the respiratory tract, caused by infection with human rhinovirus and other irritants. AB - Free ionic zinc (Zn2+) in saliva shortens duration and severity of common cold (CC) symptoms. It is proposed that Zn2+ complexes with proteins of critical nerve endings and surface proteins of human rhinovirus (HRV) (a) interrupt nerve impulses and (b) block docking of HRV on intercellular adhesion molecule-1 (ICAM 1) on somatic cells, thereby interrupting HRV infection. Since leukocyte function associated antigen-1 (LFA-1) binds leukocytes to cells through ICAM-1, initiating inflammation, Zn2+ is expected to block LFA-1/ICAM-1 binding and thereby suppress inflammation. This could explain reduction of inflammation experienced by persons taking zinc gluconate/glycine (ZGG) lozenges for CC. Allergic rhinitis (AR) and CC share many common symptoms, and ZGG also mitigates AR symptoms. Focal irritation, increased ICAM-1 expression, and recruitment of leukocytes to epithelial foci are the common elements. Zinc ions may be an important anti inflammatory factor because they can block docking of both HRV and LFA-1 with ICAM-1. PMID- 9352507 TI - Laser tweezers in cell biology. Introduction. PMID- 9352508 TI - Forces of a single-beam gradient laser trap on a dielectric sphere in the ray optics regime. AB - We calculate the forces of single-beam gradient radiation pressure laser traps, also called "optical tweezers," on micron-sized dielectric spheres in the ray optics regime. This serves as a simple model system for describing laser trapping and manipulation of living cells and organelles within cells. The gradient and scattering forces are defined for beams of complex shape in the ray-optics limit. Forces are calculated over the entire cross-section of the sphere using TEM00 and TEM*00 mode input intensity profiles and spheres of varying index of refraction. Strong uniform traps are possible with force variations less than a factor of 2 over the sphere cross-section. For a laser power of 10 mW and a relative index of refraction of 1.2, we compute trapping forces as high as approximately 1.2 x 10( 6) dynes in the weakest (backward) direction of the gradient trap. It is shown that good trapping requires high convergence beams from a high numerical aperture objective. A comparison is given of traps made using bright field or differential interference contrast optics and phase contrast optics. PMID- 9352509 TI - Basic laser tweezers. AB - The basic information has been provided here for designing and building a laser tweezers system for force measurements. If force measurements are not required, then the considerations about the analysis system, a fine piezo stage, and stability are less important. For the initial alignment and characterization of the system, red blood cells provide an easily trapped sample. For a difficult test sample, the smaller latex beads (0.15-0.3 micron in diameter) are stable and easy to obtain. Anyone setting up laser tweezers is encouraged to see a working tweezers system and to compare samples with that system. Everyone has a different background, and there may be aspects critical for you that have not been discussed here. More sophisticated systems are described later in this book. PMID- 9352510 TI - A simple assay for local heating by optical tweezers. PMID- 9352512 TI - Laser scissors and tweezers. AB - In summary, we described the use of laser scissors and tweezers from three perspectives: (a) the historical background from which these two techniques evolved, (b) an understanding and lack of understanding of the mechanisms of interaction with the biological systems, and (c) the applications of the scissors and tweezers alone and in combination. As the technology improves and we gain a better understanding of how these two tools operate they will become even more useful in probing cell structure and function, as well as practically manipulating cells in genetics, oncology, and developmental biology. PMID- 9352511 TI - Reflections of a lucid dreamer: optical trap design considerations. PMID- 9352513 TI - Optical force microscopy. PMID- 9352514 TI - Single molecule imaging and nanomanipulation of biomolecules. PMID- 9352515 TI - Signals and noise in micromechanical measurements. PMID- 9352516 TI - Cell membrane mechanics. PMID- 9352517 TI - Application of laser tweezers to studies of the fences and tethers of the membrane skeleton that regulate the movements of plasma membrane proteins. PMID- 9352518 TI - In vivo manipulation of internal cell organelles. PMID- 9352519 TI - Optical chopsticks: digital synthesis of multiple optical traps. PMID- 9352520 TI - Diagonal neglect on cancellation. AB - Patients with right hemisphere injury frequently neglect to cancel targets primarily in the left part of the page nearest the body. Since this region is diagonally opposite the area from where such patients usually begin cancelling, near left ('diagonal') neglect may be consequent to stimulus order effects ('fatigue'). We evaluated the persistence of near left neglect in nine stroke patients when they had to cancel either the near or the far half of the page before proceeding to the other half. Our results showed that near left neglect on the page was largely unaffected by cancellation order. Furthermore, a near left gradient of omissions was established within both radial (near and far) halves of the page, as well as for the entire page. Our findings suggest that diagonal cancellation neglect is unrelated to fatigue and reflects a consistent, two dimensional disorder of spatial attention. Such neglect may be related to the extent of the visible stimulus array, as well as to the array's location in egocentric space. PMID- 9352521 TI - Neuroanatomical correlates of pleasant and unpleasant emotion. AB - Substantial evidence suggests that a key distinction in the classification of human emotion is that between an appetitive motivational system association with positive or pleasant emotion and an aversive motivational system associated with negative or unpleasant emotion. To explore the neural substrates of these two systems, 12 healthy women viewed sets of pictures previously demonstrated to elicit pleasant, unpleasant and neutral emotion, while positron emission tomographic (PET) measurements of regional cerebral blood flow were obtained. Pleasant and unpleasant emotions were each distinguished from neutral emotion conditions by significantly increased cerebral blood flow in the vicinity of the medial prefrontal cortex (Brodmann's area 9), thalamus, hypothalamus and midbrain (P < 0.005). Unpleasant was distinguished from neutral or pleasant emotion by activation of the bilateral occipito-temporal cortex and cerebellum, and left parahippocampal gyrus, hippocampus and amygdala (P < 0.005). Pleasant was also distinguished from neutral but not unpleasant emotion by activation of the head of the left caudate nucleus (P < 0.005). These findings are consistent with those from other recent PET studies of human emotion and demonstrate that there are both common and unique components of the neural networks mediating pleasant and unpleasant emotion in healthy women. PMID- 9352522 TI - Impaired visual search in patients with unilateral neglect: an oculographic analysis. AB - The attentional deficit underlying hemispatial neglect was examined through a detailed analysis of the eye movement performance of a group of neglect patients. Relative to normal subjects and to patients with hemianopia without neglect, patients with left neglect make fewer fixations and have shorter inspection time on the contralesional left side. They also start their search to the right of the midline and make significantly more fixations and longer fixations on the ipsilesional right side. A positive linear relationship between horizontal location and frequency of fixations was noted for the neglect group as a whole, as well as for most of the individual patients. These findings strongly endorse the view that the attentional deficit in neglect follows a left right gradient. The peak of the maximum fixations, however, is not on the extreme right, as might be predicted by a strict gradient account, and is more consistent with recent views that the midsagittal plane of the viewer is redirected rightwards. These findings provide a detailed analysis of the eye movement patterns in neglect patients and demonstrate the robustness of oculographic analysis for examining their altered spatial representation. PMID- 9352523 TI - Comparing the visual deficits of a motion blind patient with the visual deficits of monkeys with area MT removed. AB - The performance of a 'motion blind' patient on a series of tasks in which the perception of motion played an essential or no role was compared with that of a human subject with normal vision and with that of macaque monkeys in which cortical visual area MT had been removed and adjacent areas damaged. The patient experienced difficulties on those tasks in which the perception of motion was essential, but was unimpaired on those tasks that did not require it. Similarly, the tasks which the 'motion blind' patient found impossible or difficult were precisely those tasks on which monkeys lacking area MT performed poorly. Similarly, the tasks on which the patient performed well also presented no difficulties for the animals lacking cortical area MT. The close correlation between the pattern of visual perceptual impairments in the patient and monkeys indicates that the patient's inability to perceive most forms of visual movement is attributable to total loss of, or extensive damage to, a cortical visual area that is the human equivalent of area MT and perhaps its adjacent areas. PMID- 9352524 TI - Laterality effects in the processing of melody and timbre. AB - Laterality for the processing of melody and timbre was investigated in 64 right handed non-musicians. In one block of dichotic-listening trials, participants listened for a prespecified target melody, and in a second block they listened for a prespecified target instrument. Females were more accurate on the left ear in the melody task (whereas males tended to show no ear advantage), but there were no significant ear differences in the timbre task for either sex. This supports the idea of a complementary sex-based pattern of lateralization, with males more strongly lateralized for verbal stimuli and females more strongly lateralized for non-verbal stimuli. No relation was observed between lambda measures for the two tasks, suggesting that laterality for melody processing is independent of laterality for timbre processing. PMID- 9352525 TI - Response bias affects perceptual asymmetry scores and performance measures on a dichotic listening task. AB - A dichotic listening paradigm discussed by Sidtis and Bryden (Neuropsychologia, 1978, 16, 627-632) allows one to present non-verbal as well as verbal material. This paradigm also permits signal-detection analyses to separate response biases from discrimination abilities. The present study used Sidtis' (Neuropsychologia, 1981, 19, 103-112) Complex Tone Task as an example of the paradigm. Employing signal-detection analyses, we demonstrated that commonly used performance and asymmetry indices are confounded by response bias. Several indices based on signal-detection measures are suggested to replace current widely used measures. As pointed out by Bryden and Sprott (Neuropsychologia, 1981, 19, 571-581), currently the usefulness of a perceptual asymmetry score is mainly determined by mathematical and statistical properties rather than by a theoretical framework. Thus, the choice of a particular index based on signal-detection theory is arbitrary. The present results and those of Katsuki et al. (Journal of Speech and Hearing Research, 1984, 27, 444-448) suggest that the confounding effect of response bias may be present in a variety of experiments investigating lateral processing. PMID- 9352526 TI - Visual-imitative dissociation apraxia. AB - Liepmann posited that, in right handers, the left parietal lobe contains movement formulas or representations. Therefore, performance failures may be induced by degraded representations, a failure of these representations to influence motor systems or a failure of stimuli to fully access these representations. Imitation may help the performance of subjects with degraded representations. However, patients who have impaired visual access to movement representations may perform more poorly with imitation than to verbal command. Trajectories of repetitive 'slicing' gestures made by a previously reported subject (Raymer et al.) with an infarction in the left visual association cortex (left occipital and inferior temporal lobe) that spared the parietal lobe were contrasted with those of three apraxic subjects with lesions that included the left parietal lobe and four non brain-damaged control subjects. All subjects were asked to produce the gesture to verbal command and to imitation. Movements of the left hand, wrist, elbow and shoulder were digitized from neighboring views, reconstructed in three dimensions, and analysed graphically and numerically. The apraxic subjects with left parietal damage were unable to maintain the proper linearity and spatiotemporal attributes of their wrist motions and showed interjoint coordination deficits. Their deficits were most pronounced to verbal command, with their movements improving though remaining poorly performed when they imitated. The subject with the left occipital and inferior temporal lesion that spared parietal cortex, however, showed an opposite pattern. This subject exhibited close to normal performance when producing the movement to verbal command, but significant deficits when imitating. PMID- 9352527 TI - Mirror writing in right-handers and in left-handers: a study using Chinese characters. AB - The mechanism of mirror writing was investigated using legal Chinese characters and illegal pseudocharacters. It was found from the results of right-handers in experiment 1 that the performance of normal writing and mirror writing for legal characters was better than that of normal writing and mirror writing for pseudocharacters. A reasonable explanation for this character-superiority effect is that normal engrams and mirror engrams exist only for legal characters but not for pseudocharacters. Further analysis revealed that the character-superiority effect took place in normal writing only when the right hand was used and the same effect was observed in mirror writing only when the left hand was used. It seemed that the normal engrams used in normal writing were stored in the left hemisphere while the mirror engrams used in mirror writing were stored in the right hemisphere. The results in experiment 2 from the left-handers showed the similar pattern as that of the right-handers. The mirror-engram hypothesis seems to be the best mechanism to account for the performance difference of right hand and left hand in mirror writing. PMID- 9352528 TI - Image generation and handedness: is the hemi-imagery method valid for studying the hemisphere imagery generation process? AB - The validity of the proposal by Shuren et al. (Neuropsychologia, 1996, 34, 491 492) that there is some relation between handedness and hemi-imagers was investigated by means of the hemi-imagery test. Two-hundred and two subjects were asked to image one-half of an object and to report which half (left or right) they saw. The analyses, based on criteria identical to those of Shuren et al., did not replicate their findings that subjects were more likely to image the right half of objects than the left, and that right handers are right hemi imagers. Rather, our subjects were more inclined to image the left half than the right, and no relation between handedness score and right hemi-imagers was found. It is suggested that the reading habits of the different cultures of the groups of subjects may account for this discrepancy. PMID- 9352529 TI - Patterns of dissociation between left hemineglect and deviation of the egocentric reference. AB - Sixteen control subjects and six right brain-damaged patients with left hemiparesis (three showing signs of left unilateral neglect, three with no signs of neglect) performed a straight-ahead pointing task with their right hand while blindfolded. The aim was to test the hypothesis that the egocentric reference shows significant ipsilesional deviation in left neglect patients. We found no correlation between the position of the egocentric reference and the presence of neglect signs. Neglect patients, like non-neglect patients, showed leftward, rightward or no significant deviation when pointing straight ahead. Results are discussed with reference to egocentric hypotheses of neglect and experimental remission of neglect. PMID- 9352530 TI - No evidence of hemispheric facilitation following the induction of negative and positive affect. AB - A dichotomy is thought to exist between the hemispheres whereby the right hemisphere is specialized for the processing of negative emotions and the left hemisphere is specialized for positive emotions. Van Strien and Morpurgo (Neuropsychologia, 1992, 30, 845-848) demonstrated that the activation of negative and positive emotional states resulted in the allocation of attentional resources to the contralateral hemispace. In the present experiment we sought to replicate this effect and improve upon certain methodological features of the experiment. The effect of positive and negative emotions on performance in the left and right visual fields was investigated in 30 dextral students. Positive and negative emotional states were generated by presenting subjects with an emotive word prior to each trial and subsequently requiring them to use that word within a sentence. Performance within the left and right fields was measured using a gap detection task which was neutral in relation to functional asymmetry. No evidence of right or left visual field facilitation was found for the positive or negative conditions, respectively. These results are not interpreted as a refutation of hemispheric specialization for emotional valence. Instead, they are seen to highlight the frailty of hemispheric facilitation effects. PMID- 9352531 TI - In vivo radiotracers for vesicular neurotransmitter transporters. AB - The vesicular monoamine transporter is a specific presynaptic protein involved in the transport of monoamines from the cytosol to storage vesicles of monoaminergic nerve terminals. Recently, radioligands for this transporter have been developed and utilized for in vivo positron emission tomographic (PET) imaging of monoaminergic nerve terminals in the human brain. In this review, the characteristics of vesicular transport and storage that provided the impetus for development of these radioligands are presented and discussed. PMID- 9352532 TI - [18F] beta-CIT-FP is superior to [11C] beta-CIT-FP for quantitation of the dopamine transporter. AB - beta-CIT-FP [N-(3-fluoropropyl)-2 beta-carbomethoxy-3 beta-(4 iodophenyl)nortropane] is a cocaine analogue with high affinity for the dopamine transporter. Positron emission tomography (PET) studies with [O-methyl-11C] beta CIT-FP ([11C] beta-CIT-FP) has shown that equilibrium conditions were approached but, however, not reached at the end of measurement. Moreover, metabolite studies of [11C] beta-CIT-FP in monkey plasma demonstrated a lipophilic-labelled metabolite that may enter the brain. We therefore labelled beta-CIT-FP with fluorine-18 in a position that may avoid the formation of labelled lipophilic metabolites. The more long-lived radionuclide (18F) was used to allow for measurements over longer time. [N-fluoropropyl- 18F] beta-CIT-FP ([18F] beta-CIT FP) was prepared by N-alkylation of nor-beta-CIT with [18F]fluoropropyl bromide. PET studies were performed in cynomolgus monkeys. [18F] beta-CIT-FP entered the brain rapidly. There was a high concentration of radioactivity in the striatum and much lower in the thalamus, neocortex, and cerebellum. The striatum-to cerebellum ratio was about 5 at time of transient equilibrium, which occurred after 60 to 100 min. After pretreatment with GBR 12909, radioactivity in the striatum was markedly reduced, thus indicating specific [18F] beta-CIT-FP binding to the dopamine transporter. The fraction of unchanged [18F] beta-CIT-FP determined by HPLC was 10-15% after 140 min. No lipophilic labelled metabolites were detected. The absence of measurable lipophilic labelled metabolites and the occurrence of transient equilibrium within the time of the PET measurement indicate that [18F] beta-CIT-FP is superior to [11C] beta-CIT-FP as a PET radioligand for quantification of the dopamine transporter in the human brain. PMID- 9352533 TI - [125I] beta-CIT-FE and [125I] beta-CIT-FP are superior to [125I] beta-CIT for dopamine transporter visualization: autoradiographic evaluation in the human brain. AB - The binding of the three dopamine transporter radioligands ([125I] beta-CIT, [125I] beta-CIT-FE, and [125I] beta-CIT-FP) was studied using whole-hemisphere autoradiography on postmortem human brains. The autoradiograms revealed an intense and homogeneous labeling of the nucleus caudatus and putamen but also to varying extent to serotonergic and noradrenergic transporters of neocortex and thalamus. The order of specificity estimated (striatum over neocortex ratios) was beta-CIT-FP > beta-CIT-FE > > beta-CIT, suggesting that beta-CIT-FE and beta-CIT FP should be preferred for in vivo studies of the dopamine transporter in the human brain. PMID- 9352534 TI - Importance of pre-treatment radiation absorbed dose estimation for radioimmunotherapy of non-Hodgkin's lymphoma. AB - Non-Hodgkin's lymphoma I-131 radioimmunotherapy data were analyzed to determine whether a predictive relationship exists between radiation absorbed doses calculated from biodistribution studies and doses derived from patient size. Radioactivity treatment administrations scaled to patient size (MBq/kg or MBq/m2) or fixed MBq doses do not produce consistent radiation absorbed dose to critical organs. Treatment trials that do not provide dose estimates for critical normal organs are less likely to succeed in identifying a clinical role for radioimmunotherapy. PMID- 9352535 TI - In vitro and in vivo behavior of radiolabeled chimeric anti-EGFRvIII monoclonal antibody: comparison with its murine parent. AB - The mutant version of the epidermal growth factor receptor EGFRvIII has been found on gliomas and other tumors, but not on normal tissues. Radioiodinated murine (mu) L8A4 monoclonal antibody (MAb) specifically targets EGFRvIII xenografts in vivo when labeled using N-succinimidyl 5-iodo-3-pyridinecarboxylate (SIPC). A chimeric (ch) MAb consisting of the variable region of muL8A4 and the constant domains of human IgG2 has been developed that has an affinity and radioiodinated immunoreactive fraction comparable to muL8A4. In vitro, both MAbs were internalized and processed by EGFRvIII expressing cell lines (U87MG delta EGFR or NR6M) at similar rates (maximum intracellular retention, 35-40%). In paired-label tissue distribution studies in athymic mice bearing U87MG delta EGFR tumor xenografts, the ch:mu L8A4 uptake ratio in normal tissues rose to greater than 2:1, whereas in tumor, the ratio remained 1:1 throughout the experiment. These results indicate that chL8A4 exhibits similar binding and internalization properties as its murine parent, but suggest different intracellular processing and/or deposition of catabolites in normal tissues for chL8A4. PMID- 9352536 TI - Detection of experimental infections with 99mTc-labeled monoclonal antibodies against TNF-alpha and interleukin-8. AB - This study was designed to assess monoclonal antibodies (MAbs) directed against tumor necrosis factor-alpha (TNF-alpha) (anti-TNF) or interleukin-8 (anti-IL-8) as radioactive agents for the detection of Staphylococcus aureus-or Klebsiella pneumoniae-infected thighs in mice. At 5 min (acute infection) or 20 h (established) post-infection, 20 micrograms of the 99mTc-labeled MAbs were injected. At various time intervals, the accumulation of the radiotracer in the infected thighs was assessed and expressed as a target-to-nontarget (T/NT) ratio. The binding of 99mTc-labeled MAbs to circulating mononuclear cells and granulocytes was quantitated 20 h after injection. The pharmacokinetics of the MAbs, in relation to the control agents 99mTc-labeled polyclonal human immunoglobulin (IgG) and a 99mTc-labeled nonspecific IgG1 MAb, were also studied. In acute infections, 99mTc-anti-TNF accumulated to a higher extent (p < 0.05) in S. aureus-infected thighs in mice until 4 h after the injection than 99mTc-IgG and was higher at 0.25 h in K. pneumoniae-infected mice (p < 0.03) compared with 99mTc-IgG. In established S. aureus and K. pneumoniae infections, 99mTc-anti-IL-8 detected the infection more intensely than 99mTc-IgG until 1 h after injection. In both S. aureus and K. pneumoniae infections, localization of sites of infection correlates (p < 0.05) with increased binding of the 99mTc-labeled MAbs to granulocytes and mononuclear cells in both acute and established infections. It was concluded that 99mTc-labeled MAbs, directed against TNF-alpha and IL-8, accumulate in bacterial infections in mice to a higher extent than does 99mTc-IgG after infection and is related to the binding of the antibodies to blood leukocytes. With these 99mTc-labeled MAbs, information might be gained about the development of an infection. PMID- 9352537 TI - Intratumoral microdistribution of [131I]MB-1 in patients with B-cell lymphoma following radioimmunotherapy. AB - Intratumoral microdistribution of radiolabeled anti-CD37 murine monoclonal antibody, [131I]MB-1, in lymph nodes from five patients with non-Hodgkin's B-cell lymphoma following radioimmunotherapy were evaluated by microautoradiography and image analysis of macroautoradiographs. Microdistribution of radioactivity was highly heterogeneous: silver grain counts varied from 28-70 to 8-10 per 400 X field, and the coefficients of variations calculated by image analysis ranged between 42.5 and 79.3%. Variable radiation doses delivered could have contributed to the limited durability of tumor regression. PMID- 9352538 TI - Reassessment of FDG uptake in tumor cells: high FDG uptake as a reflection of oxygen-independent glycolysis dominant energy production. AB - To determine appropriate use of 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) in the diagnosis of malignant tumors, the mechanism of enhanced FDG uptake in tumor cells was reassessed using in vitro cultured cell lines and 3H-deoxyglucose (DG), in combination with possible parameters of aerobic and anaerobic energy production. The high DG uptake in the tumor cells reflected the dependency of energy production on anaerobic glycolysis, and paradoxically on low levels of aerobic oxidative phosphorylation in mitochondria. We discuss here factors underlying anaerobic glycolysis in tumor cells. PMID- 9352540 TI - Fluorine-18-labeled fluorine gas for synthesis of tracer molecules. AB - The aim of this work was to develop a method to produce 18F-labeled fluorine gas ([18F]F2) with high specific radioactivity (SA, radioactivity/mass-ratio). 18F Labeled methyl fluoride ([18F]CH3F) was synthesized from [18F]F-aq and mixed with carrier F2 in an inert neon matrix. The constituents were atomized in an electric discharge, after which a rearrangement and 18F for 19F exchange took place. [18F]F2 with a specific radioactivity of up to 55 GBq/mumol is available for the labeling synthesis of tracers for positron emission tomography (PET). PMID- 9352539 TI - N-(N-benzylpiperidin-4-yl)-2-[18F]fluorobenzamide: a potential ligand for PET imaging of sigma receptors. AB - Four nitro- and fluorobenzamides (1-4) have been synthesized in good yields from nitro- and fluoro-substituted benzoyl chloride with 4-amino-1-benzylpiperidine. In vitro studies showed that these compounds have high affinities to sigma receptors. N-(N-Benzylpiperidin-4-yl)-2-fluorobenzamide (3), in particular, bound to sigma receptors with high affinity (Ki = 3.4 nM, guinea pig brain membranes) and high selectivity (sigma-2/sigma-1 = 120). It was, therefore, labeled with 18F and evaluated as a sigma receptor radioligand. N-(N-Benzylpiperidin-4-yl)-2 [18F]fluorobenzamide (3a) was synthesized in one step by nucleophile substitution of the 2-nitro precursor (1) with [18F]fluoride in DMSO at 140 degrees C for 20 min followed by purification with HPLC in 4-10% yield (decay corrected). The synthesis time was 90 min and the specific activity was 0.4-1.0 Ci/mumol. Tissue distribution in mice revealed that the uptakes of 3a in the brain, heart, liver, lungs, spleen, kidneys and small intestine were high, and the radioactivity in these organs remained constant from 60 to 120 min post-injection. The radioactivity in the bone did not significantly increase, suggesting in vivo defluorination may not be the major route of metabolism of 3a in mice. Blocking studies with haloperidol in rats indicated that the uptake of compound 3a in the rat brain was selective to haloperidol-sensitive sigma sites. These results suggest that compound 3a is a potent sigma receptor radioligand and may be a potential ligand for PET imaging of sigma receptors in humans. PMID- 9352541 TI - In vitro and in vivo characterization of novel water-soluble dithio-bisphosphine 99mTc complexes. AB - The water-soluble dithio-bis(hydroxymethyl)phosphine ligands (HOH2C)2P(CH2)2 S(CH2)3S(CH2)2P(CH2OH)2 and (HOH2C)2P(CH2)3S(CH2)3S(CH2)3P(CH2OH)2 were complexed with 99mTc. The 99mTc-P2S2 complexes were formed in high radiochemical purity by simple mixing of 99mTcO-4 with the ligands or by transchelation from 99mTc citrate. The 99mTc-P2S2 complexes were stable over a wide range of pHs and did not undergo in vitro decomposition for < or = 24 h. High performance liquid chromatographic analysis indicated the formation of singular chemical species. Retention times for each of the new 99mTc-P2S2 complexes are identical to those of corresponding Re(V) complexes, suggesting similar chemical species at the tracer level. Results of this study suggest that the combination of thioether and (hydroxymethyl)phosphine donor centers in new, multidentate ligand frameworks might aid in the development of new bifunctional chelating agents for the use of radiolabeling specific biomolecules. PMID- 9352542 TI - Regional cerebral blood flow measured by microsphere technique in experimental animals: technical notes for the use albumin microspheres in rats. PMID- 9352543 TI - Formulation and evaluation of a two-components lyophilized kit for Tc-sestamibi: transchelation preparation of Tc-99m-sestamibi. PMID- 9352544 TI - 3D-mode acquisition in clinical PET. PMID- 9352545 TI - Classification of mild Alzheimer's disease by artificial neural network analysis of SPET data. AB - An evaluation of the performance of artificial neural networks (ANNs) for the classification of probable Alzheimer's disease (pAD) patients was undertaken using data extracted from four regions of interest constructed on single photon emission tomographic (SPET) cerebral perfusion images. Two studies using feed forward neural networks (FFNNs) were undertaken. The first was to determine if it would be possible to classify pAD patients and normal subjects in a mixed group, comprising 29 patients diagnosed as having pAD varying in severity from mild, established dementia to moderate dementia and 10 healthy control subjects. The second was to determine if the networks generated in the first study could prospectively classify 15 additional patients with very mild or mild cognitive impairment. The results were compared to those obtained using the same data and discriminant analysis. The relative performances of the two analysis techniques were assessed on the basis of the area under receiver operating characteristics (ROC) curves. The FFNN successfully classified all datasets in the first study, achieving an area under the ROC curve of 1.00, whereas discriminant analysis achieved 0.94. When tested on data from the second group, the areas under the ROC curves varied between 0.86 and 1.00 for the FFNN, whereas that for discriminant analysis was 0.99. We conclude that FFNNs can accurately classify pAD patients with mild to moderate dementia using data obtained from SPET cerebral perfusion images. PMID- 9352546 TI - Quantification of IBZM dopamine receptor SPET in de novo Parkinson patients before and during therapy. AB - A number of neurodegenerative diseases have been evaluated with 123I iodobenzamide (123I-IBZM) dopamine receptor scintigraphy, including Parkinson's disease. Differential diagnosis is based on the semi-quantitative determination of striatal uptake in the basal ganglia. Seven procedures for calculating basal ganglia uptake were compared and checked statistically in (1) 28 previously untreated de novo parkinsonian patients before and (2) 14 patients after (mean of 9 months) commencement of anti-Parkinson medication. Of the 21 hemi-parkinsonian patients, 16 demonstrated increased uptake contralaterally (mean right-to-left difference = 8%, sensitivity = 76%) using the most robust uptake procedure. The difference in uptake between the affected and contralateral sides (mean = 6%) was significant (P = 0.02). The mean (+/- S.D.) basal ganglia/frontal cortex (BG/FC) ratio was 1.55 +/- 0.14 (attenuation-corrected). Attenuation correction did not affect the relative ratio of basal ganglia uptake (P = 0.01). The anti-Parkinson medication did not result in any significant changes in the BG/FC ratio at follow up, but responders could be differentiated from non-responders based on initial uptake (mean BG/FC ratio of 1.58 and 1.39 respectively). We conclude that 123I IBZM can be used routinely to identify which Parkinson patients will benefit from dopaminergic medication. PMID- 9352547 TI - Assessment of drainage in PUJ dilatation: pelvic excretion efficiency as an index of renal function. AB - Renal function is important when assessing the response of a dilated renal pelvis to a diuretic stimulus. Yet there is little in the literature to suggest how this should be undertaken. Our aim was to develop a model which we have called pelvic excretion efficiency (PEE). The PEE, which may be used to assess drainage, is a mathematical model of the ratio of the total kidney excretion to the total amount of isotope extracted from the blood by the kidney. Thirty-three children with a prenatal diagnosis of unilateral renal pelvic dilatation (PUJ) were treated conservatively after birth. As a group, they underwent a total of 164 diuretic DTPA renograms up to the age of 72 months. Drainage was assessed as the response to frusemide (defined as the time for the corrected renal curve to fall to 75% of the maximum value in the frusemide part of the study; T75), response to bladder emptying, a change of posture after frusemide (PM), and PEE. The contralateral normal kidney showed a combination of both 'good' T75 and PM drainage in 51% of renograms while the PEE showed drainage in all. The affected kidney with renal pelvic dilatation showed a combination of both 'poor' T75 and PM drainage in 42% of renograms. The PEE was low in 99% of these 'poor drainage' renograms. The PEE, the ratio of the mathematical model of renal uptake to excretion, is readily calculated and may be a more accurate and specific technique to assess drainage on diuretic renography. PMID- 9352548 TI - A follow-up study of vesico-ureteric reflux and renal scars in asymptomatic siblings of children with reflux. AB - The objectives of this study were to follow up children with vesico-ureteric reflux (VUR) and renal scars, to evaluate kidney growth and to determine the incidence of urinary tract infection (UTI) and elevated blood pressure in 40 asymptomatic siblings of children with VUR, in whom VUR had been detected at an early age, and to gather additional data which could help to evaluate the need for screening for VUR in asymptomatic siblings. During the follow-up period of 3 7 years, two children (5%) had UTI; 66% of VUR grade 1 and 2 disappeared. The progression of scars was only detected in two of nine children with renal scars on the initial study, both of whom had high-grade VUR. Renal ultrasound was normal in all siblings and none developed hypertension. The results indicate that low-grade sterile VUR may not play a major role in renal scarring, but this may not be the case with high-grade sterile VUR. Considering the correlations among VUR, UTI and reflux nephropathy, routine screening for VUR at an early age in asymptomatic siblings of children with VUR seems to be justified to identify those at the greatest risk of subsequent renal damage. PMID- 9352549 TI - Renal vascular transit time and tubular transit time dispersion for 99Tcm-MAG3. AB - Renal transit time usually refers to tubular transit time, as introduced by Taplin, but other measures of renal transit have been proposed. Here we examine the vascular transit time (VTT, following Rutland) and the standard deviation of tubular transit time (SDTT, following Britton) in a group of 30 patients having baseline and ACE-inhibitor 99Tcm-MAG3 renography prior to arteriography. A same day, low-dose/high-dose protocol was used for renography; only the post-captopril dose was high enough to measure VTT. Pre-captopril, the Spearman rank correlation coefficient for SDTT was rho = 0.52 (n = 53 kidneys; P < 0.0002); post-captopril, rho = 0.54 (n = 49 kidneys; P < 0.0002). For VTT, the post-captopril value was rho = 0.24 (n = 30 kidneys; N.S.). For comparison, the same statistics were calculated for Taplin's original measure of transit time: the time from injection to maximum count rate (peak time). Pre-captopril, for peak time, rho was 0.47 (n = 53 kidneys; P < 0.001); post-captopril, rho was 0.39 (n = 50 kidneys, P < 0.01). These findings confirm the diagnostic value of SDTT but not of VTT. SDTT correlated better than peak time with the arteriographic findings. PMID- 9352550 TI - 99Tcm-sestamibi scintimammography in patients with suspicious breast lesions: comparison of SPET and planar images in the detection of primary tumours and axillary lymph node involvement. AB - Planar scintimammography with 99Tcm-sestamibi (99Tcm-MIBI) has been shown to be useful in diagnosing breast carcinoma. The aim of this study was to compare single photon emission tomography (SPET) and planar imaging for scintimammography with 99Tcm-MIBI in the detection of primary breast cancer and axillary lymph node involvement. Sixty-three females with mammographically suspicious lesions and 12 controls were evaluated. Dynamic images were acquired commencing immediately after the injection of the radiopharmaceutical, followed by multiple planar images in the supine and prone positions plus SPET supine imaging. A final histopathological diagnosis was achieved after surgery. A total of 66 breast lesions were considered. No focal uptake of 99Tcm-MIBI was observed in the breasts or axillas of the controls. In the patients with breast cancer, the sensitivity was 92.9% (39/42) for SPET, 71.4% (30/42) for supine and 85.7% (36/42) for prone planar imaging, respectively; the specificity was 87.5% for SPET and 91.6% for the planar scans. Metastatic axillary lymph node involvement was seen in 19 patients: the sensitivity was 84.2% (16/19) for SPET and 63.2% (12/19) for planar images; the specificity was 91.3% and 95.7% respectively. Our results confirm the high diagnostic accuracy of 99Tcm scintimammography in the diagnosis of breast cancer, and suggest that SPET is more sensitive than planar images, especially in detecting axillary lymph node involvement. PMID- 9352551 TI - 99Tcm-MDP scintigraphy in high-voltage electrical burn patients. AB - In high-voltage electrical burn injuries (> 1000 V), it is difficult to identify the site and extent of non-viable deep tissue damage for debridement to avoid further tissue injury from wound infection and the risk of sepsis. This prospective study was designed to evaluate the usefulness of 99Tcm-methylene di phosphonate (99Tcm-MDP) scintigraphy in detecting the extent of tissue injury and determining the level of amputation required for electrical burn patients. Over a 5 year period, 33 high-voltage electrical burn patients were studied. Blood flow and blood pool studies revealed absent perfusion in 37 limbs, all of which eventually were amputated. In addition to a routine three-phase bone scan, images were obtained at 30-60 min (early images) to evaluate whether soft tissue injury could be detected better at that time. For comparison of the detection rate from the early images and bone (delayed) images, 164 corresponding spot views of both images were reviewed. Eighty-three and 125 tissue necrotic lesions were demonstrated by the early images and bone images respectively. All of the 83 lesions found by the early images were more clearly identified by the bone images. All but one of the 125 lesions underwent surgical debridement or amputation. We concluded that the blood flow and blood pool images correlated well with the level of amputation required. The site and extent of tissue necrotic lesions can be clearly identified on 99Tcm-MDP bone scans. Because the early images were less sensitive in detecting tissue necrosis, we suggest that early imaging is not necessary. PMID- 9352552 TI - The role of bone scintigraphy and plain radiography in intractable plantar fasciitis. AB - The objective of this study was to assess the role of bone scintigraphy and plain film radiography in intractable plantar fasciitis. The bone scintigrams, radiographs and clinical histories of 33 patients with chronic plantar fasciitis were reviewed. These patients were refractory to conservative treatments and were being considered for surgical plantar fascia release. Twenty-eight patients had increased uptake on scintigraphy at the medial calcaneal tubercle, while a plantar spur was seen in 21 patients. Seventy-five percent of patients with increased uptake had a calcaneal spur; 95% of patients with a spur had increased uptake on scintigraphy. It would appear that plantar calcaneal spurs are more prevalent in this group of patients than in the general population and, although they may not be the primary cause of pain, they may predispose to it. Scintigraphy was helpful in patients without a spur or with atypical symptoms or signs. It did not provide any further information on the group of patients with a spur. PMID- 9352553 TI - Out-of-hours weekend scintigraphy: assessing/predicting the need. AB - We have assessed the potential impact of a regular half-day session on Saturday only, or Sunday only, and compared this with a whole weekend on-call service for lung scans. We predicted the effect of these services using the data gathered over 2 years (1992-94), looking at the results of lung scans and admission and discharge of patients. The on-call service in all three cases would be justified if resources from the savings on patient discharge and bed availability could be earmarked for the nuclear medicine service. The cost of introducing such an on call service for the department would be Pounds 3000 per year per session at the weekend and up to Pounds 10,000 per year for a full weekend on-call service. The total cost to the hospital would be negligible. PMID- 9352554 TI - Computer simulations of lung morphologies within planar gamma camera images. AB - A mathematical model and computer code have been developed to unambiguously interpret planar gamma camera images. Specifically, the algorithm permits airway composition of the central, intermediate and peripheral partitions of scans to be determined quantitatively. The algorithm unambigously identifies the spatial coordinates of each of the millions of airways within the adult human lung, and assigns every individual airway to a precise location within the gamma camera image format prescribed by the clinician. This is done on a patient-by-patient basis. The algorithm has evolved from clinical applications of the previous work of Martonen et al. [1]. The objective of the current work was to advance the original protocol and derive an algorithm that was: (1) from a medical perspective, more physiologically realistic; and (2) from a technical perspective, easier to apply in the medical arena. Regarding (1), the major elements of the algorithm are that the outer boundary of the lung is formulated directly from lung perfusion imaging data and that the lung is divided into distinct left and right components. Regarding (2), the algorithm has been written for use with common workstations. It is our hope that the improvements will facilitate applications of the new model-code into aerosol therapy regimens. PMID- 9352555 TI - Uptake of 99Tcm-nitrido dithiocarbamate complexes by tumour cells. AB - Uptake of radiopharmaceuticals by tumour cells may provide useful information on the biochemical characteristics of the cell, such as its drug resistance status. We have prepared a series of 99Tcm-nitrido dithiocarbamate complexes of the type [99TcmN(dtc)2] (dtc = N-R1-N-R2-dithiocarbamato, R1, R2 = Me, Me; Et, Et; Et, n Bu; Me, CH2CH2NMe2; Me, CH2CH2NMe3+; Me, CH2COOMe), and investigated the kinetics of uptake of these complexes in several tumour cell lines. The 99Tcm-nitrido dithiocarbamate complexes were prepared by stannous reduction of [99Tcm]pertechnetate in a solution of DPTA and succinic dihydrazide followed by addition of the appropriate dithiocarbamate. The complexes were analysed by reverse-phase HPLC. The complexes were incubated with the human tumour cell lines MKN-45, H-69, H-348 and MCF-7 and with normal mixed leukocytes and erythrocytes, and the uptake and washout of the various complexes were determined at various time points. Uptake was rapid, high in some cell lines and lower in others. In general, the more lipophilic complexes showed high uptake, but the most lipophilic did not show the greatest uptake. Uptake was temperature-independent. Most of the bound activity was retained by the cells after removal of unbound tracer. The mechanism of uptake is different to that of 99Tcm-MIBI. The high and stable uptake of these complexes suggests that they may have application for in vivo tumour imaging and characterization, and further studies are required to establish their mechanisms of uptake. PMID- 9352556 TI - Pharmacokinetics and radiation dosimetry of 99Tcm-labelled monoclonal antibody B43.13 in ovarian cancer patients. AB - OVAREX MAb B43.13 is a new radiopharmaceutical based on a monoclonal antibody (MAb-B43.13) known to recognize CA 125, a tumour antigen associated with epithelial ovarian cancer. This MAb is capable of facile radiolabelling with 99Tcm and has been shown previously to localize in the tumours of ovarian cancer patients. The present study was initiated to measure the pharmacokinetics of this MAb in the serum of 10 patients with primary or metastatic ovarian cancer. A two compartment model was found to be best at representing the biodistribution of the 99Tcm-labelled MAb, yielding a 2.6 h distribution phase half-life and a 31.3 h elimination phase half-life. The serum and renal clearances for 99Tcm-MAb-B43.13 were 121 and 53 ml h-1 respectively. These parameters were compared with a similar model developed from the serum values of the MAb itself (determined using an ELISA detection method). Based on the serum pharmacokinetics of 99Tcm-MAb B43.13 and whole-body planar gamma camera images, an estimate of the radiation dose from 99Tcm was calculated using standard MIRD schema. The organs demonstrating significant 99Tcm uptake included the liver, kidneys, heart and spleen. The whole-body dose was similar to other 99Tcm-labelled MAbs. PMID- 9352557 TI - A computer database for the preparation of transport documents for radiopharmaceuticals. AB - The preparation of radiopharmaceuticals in central radiopharmacies for distribution to several hospitals is practised widely. Transport of the radiopharmaceuticals is usually by road and this activity is regulated by legislation. Two requirements of the legislation are that the consignor issues a transport document for each consignment and maintains a record of all transport of radioactive material. This paper describes a computer database that has been developed for use in a central radiopharmacy to achieve these requirements. PMID- 9352558 TI - Antibodies against some viruses of domestic animals in southern African wild animals. AB - Twenty-four species of South African wild animals were tested for the presence of antibodies against the viruses of 16 common diseases of domestic animals. Positive results were obtained for African horsesickness, equine encephalosis, equid herpes virus-1, infectious bovine rhinotracheitis, Allerton disease (Herpes mammillitis), lumpy skin disease, parainfluenza, encephalomyocarditis, bluetongue, Wesselsbron disease, bovine ephemeral fever, and Akabane disease complex. No antibodies could be demonstrated against the viruses of equine influenza, equine infectious anaemia, equine viral arteritis and Rift Valley fever. The negative results substantiate observations that the latter diseases, with the exception of equine viral arteritis, are absent in South Africa. The number of animal species found positive for a specific virus, ranged from 0-16. No antibodies were found in crocodiles and warthogs, whereas antibodies against Wesselsbron and bovid herpes virus-1 were present in 16 species. Antibodies against viruses of horses were found almost exclusively in zebras and, although elephants reacted to African horsesickness, no neutralizing antibodies against it could be demonstrated in their sera. Zebras were also found to be positive for Wesselsbron and Akabane, which are usually regarded as viruses of ruminants. Antibodies against most viruses were encountered in all vegetation zones in South Africa but, as a rule, most viruses were more prevalent in the high-rainfall zone in KwaZulu-Natal. PMID- 9352559 TI - Pathological changes in calves that died from experimental water intoxication. AB - The pathology of calves that died from experimental water intoxication was investigated. Oedema of the brain and urinary bladder, and renal damage were significant pathological findings in these calves. The findings were attributed to positive water balance in calves suffering from water intoxication. PMID- 9352560 TI - Acceptance of candidate baits by domestic dogs for delivery of oral rabies vaccines. AB - Protocols for evaluating oral rabies vaccine baits for domestic dogs were field tested in central Mexico, after which dog-food manufacturers and suppliers to the pet-food industry were advised as to potential ingredients for use in prototype dog baits. Bait-preference trials in which confined dogs were used were then undertaken, followed by field tests of free-ranging farmer-owned dogs in three towns in the Nile River Delta region of Egypt. Both confined and free-ranging dogs showed strong preferences for certain baits or bait coatings (poultry, beef tallow, cheese, egg and a proprietary product). Fish-meal polymer baits, widely used for wildlife species, were less preferred. In Egypt, a commercial dog-food meal bait coated with beef tallow and dry cheese, was consumed at a rate approaching that of a chicken-head bait. The percentage baits that were actually eaten after they had been offered to dogs, ranged from 71-96% for household dogs tested in Mexico, 65-91% for confined dogs (beagles and mixed breeds) tested in the United States, and 32-88% for farmer-owned dogs tested in Egypt. PMID- 9352561 TI - Helminth fauna of Anas undulata, Anas erythrorhyncha, Anas capensis and Anas smithii at Barberspan, South Africa. AB - Thirty-four species of gastrointestinal helminths were found in 25 Anas undulata, 21 Anas erythrorhyncha, ten Anas capensis and seven Anas smithii collected at Barberspan, South Africa. Excluding four new taxa, 11 new African records, and 14, 11, 12 and nine new host records were established for Anas undulata, Anas erythrorhyncha, Anas capensis and Anas smithii, respectively. The helminth community included 13 cosmopolitan species, seven species known only from the holarctic, one species known only from the south Pacific and 13 new or unidentified species that appear to be restricted to Africa. The infection levels of the common helminth species in the mainly carnivorous Anas capensis and Anas smithii, were generally much higher than those of species infecting Anas undulata or Anas erythrorhyncha. PMID- 9352562 TI - Assessment of bovine hoof conformation and its association with lameness, animal factors and management practices on small-scale dairy farms in Kiambu district, Kenya. AB - Digital health and conformation were assessed in 216 dairy cattle on 78 randomly selected small-scale farms. For each cow, gait was assessed and the digits examined in detail. Hoof measurements (angle and length of the dorsal hoof wall, heel depth and hoof-base area) were also made. Hoof measurements varied most between individual cattle. Dorsal angle was correlated with heel depth (r = 0.53; P = 0.001) and dorsal length (r = -0.40; P = 0.001). The hoof-base area was correlated with the dorsal length (r = 0.41; P = 0.001). There were significant breed differences in dorsal angle (P = 0.03) and dorsal length (P < 0.01). The dorsal angle was correlated with parity and body condition, while the dorsal length, heel depth and the hoof-base area were correlated with the heart girth (P < 0.01). Hoof conformation was associated with both clinical lameness and hoof lesions. A 1-cm increase in the dorsal length increased the odds of lameness by 16.9, heel erosion by 1.8, underrunning by 5.4 and overgrowth by 40 (P < 0.01). PMID- 9352563 TI - Prevalence of equine piroplasmosis in Central Mongolia. AB - Antigen for the indirect fluorescent antibody test (IFAT) was routinely prepared from infected erythrocytes from horses experimentally infected with Babesia equi and Babesia caballi. With the successful establishment of in vitro cultures of B. equi and B. caballi, it is now possible to employ culture-derived antigens in this test. In this study, in vitro-propagated B. equi- and B. caballi-infected erythrocytes were used as antigen in the IFAT. Various modifications to an established protocol had to be implemented to allow repeatable results. Cultures with 3-4% parasitized erythrocytes were found to be most suitable. As cross reactions of control sera on heterologous antigen were observed at serum dilutions of up to 1/40, a reciprocal titre of 80 was considered to be positive. In positive samples, specific fluorescence of Babesia parasites and/or erythrocyte membranes was observed. Fifteen sera from Babesia-free horses from Japan all tested negative in the IFAT. One hundred and ten field-horse sera from Central Mongolia were investigated in this study. The results indicate that both B. equi and B. caballi are endemic in horses in Central Mongolia, with 88.2% and 84.5% of horses being seropositive to B. equi and B. caballi, respectively. PMID- 9352564 TI - Effect of the South African asinine-94 strain of equine arteritis virus (EAV) in pregnant donkey mares and duration of maternal immunity in foals. AB - Clinical, virological and serological responses were investigated in five pregnant donkey mares after experimental exposure to the South African asinine-94 strain of equine arteritis virus (EAV), and the duration of maternal immunity to EAV was studied in their foals. In four intranasally inoculated mares, fever with maximum rectal temperatures of 39.1-40.7 degrees C was recorded 2-11 d after challenge. All the inoculated mares developed mild depression, and a serous ocular and nasal discharge; in three mares mild conjuctivitis was observed. The virus was recovered from the nasopharynx and from buffy-coat samples of all the mares 3-10 d, and 2-18 d post inoculation (p.i.), respectively. Seroconversion to EAV was detected on days 8-10 p.i. Peak serum-virus-neutralizing antibody titres of log10 1.8-2.4, and IgG ELISA OD values of 0.85-2.15 were recorded 2-3 weeks p.i. The in-contact (p.c.) control mare developed fever on days 15-19 post exposure, and showed mild clinical signs of equine viral arteritis similar to those observed in the inoculated mares. Seroconversion to EAV was detected in the p.c. mare on day 20 post exposure, and virus was isolated from nasal swabs and blood samples collected at the time of the febrile response and 1-3 d afterwards. None of the mares aborted. After they had given normal birth 45-128 d p.i. or after p.c. exposure, no virus could be isolated from their placentas. The concentration of EAV-neutralizing antibody in colostrum was two to eight times higher than in serum samples collected at the time of parturition. All the foals born to infected mares were clinically normal at the time of birth and throughout the subsequent 1-2 months of observation. No EAV was recovered from the buffy coat fraction of blood samples collected at birth nor from those collected on days 1, 2 and 7 after birth. Also, no virus-serum-neutralizing or IgG ELISA antibody to EAV was detected in sera collected immediately after birth before the foals started nursing. The colostrum-derived maternal antibodies against EAV gradually declined and could not be detected by either the VN test or ELISA for 2 3 months after birth. This study demonstrates that the asinine-94 strain of EAV does not cause abortion in pregnant donkey mares. Furthermore, no carrier state could be demonstrated in foals born to mares infected at the time of pregnancy. PMID- 9352565 TI - Development of the OPgun for bombardment of animal tissues. AB - A simple and inexpensive particle-bombardment device, the OPgun, was constructed for the delivery of DNA into animal tissues. This device is based on the particle inflow gun first described for plant-cell transfection. The delivery of tungsten particles into the epidermis of the mouse ear, without the use of vacuum and without causing damage to the tissue, was demonstrated. The system was also shown to be capable of inducing antibodies to a foreign gene in mice. PMID- 9352566 TI - Cryopreservation of sheathed third-stage larvae of Oesophagostomum radiatum (nodular worm of cattle). AB - Sheathed infective larvae of Oesophagostomum radiatum were successfully cryopreserved by the use of a procedure developed for hookworms. The survival rate, as assessed by motility, was 57.9% after 42 d of cryopreservation. PMID- 9352567 TI - Cryopreservation of third-stage larvae of Strongylus vulgaris (large strongyle of horses). AB - A technique for the cryopreservation of third-stage larvae of Strongylus vulgaris is described. Infective larvae of S. vulgaris were exsheathed in a 0.16% sodium hypochlorite solution and then transferred into cryotubes containing 0.09% saline. The samples were stored in the gas phase of liquid nitrogen. PMID- 9352568 TI - Genetic association study between pathological gambling and a functional DNA polymorphism at the D4 receptor gene. AB - A Spanish sample consisting of 68 Caucasian pathological gambling patients (47 males and 21 females) and 68 unaffected controls were screened by the molecular analysis of a functional DNA polymorphism in the locus for the D4 dopamine receptor gene. Our results are consistent with the existence of a significant association between genetic variants at a DRD4 gene polymorphism and pathological gambling (chi 2 = 11.82; P = 0.037). This association seems to be sex-influenced, since there was no significant association when only males were considered (chi 2 = 9.45; P = 0.09), but there was a more significant association if we only considered female subjects (chi 2 = 8.73; P = 0.033). Individuals with the longest allele (D7) were the most frequent in affected females (chi 2 = 4.50; P = 0.033). This work provides a new evidence of the implication of the dopaminergic reward pathways, now through the involvement of DRD4, in the aetiology of this impulsive disorder. PMID- 9352569 TI - Catechol-O-methyltransferase Val158Met polymorphism: frequency analysis in Han Chinese subjects and allelic association of the low activity allele with bipolar affective disorder. AB - Catechol-O-methyltransferase catalyses the O-methylation of biologically active or toxic catechols and is a major component of the metabolism of drugs and neurotransmitters such as L-dopa, noradrenaline, adrenaline, and dopamine. Human catechol-O-methyltransferase activity is an autosomal partially dominant trait and is strongly associated with a valine to methionine substitution at codon 158 of the protein. About 25% of Caucasians have low activity, 50% intermediate activity and 25% high activity as determined by either phenotypic or genotypic measurement. In black populations, the low activity allele (Met158; COMTL) is less frequent with about 7% being homozygous. Using a PCR based genotyping assay, we report that the Met158 allele is also less frequent in normal Han Chinese subjects with about 3% of the population being homozygous. Because of its role in catecholamine metabolism and several lines of evidence pointing to a locus for psychosis near the COMT gene on chromosome 22q11, we have analysed the COMT Val158Met polymorphism as a candidate susceptibility factor for bipolar affective disorder. We report an association between bipolar affective disorder and the Met158 allele (p = 0.004) and genotype (p = 0.01) in 93 affected Chinese subjects and 98 controls. We hypothesize that either the low activity allele of catechol-O methyltransferase is a risk factor for bipolar affective disorder in Chinese populations or is in linkage disequilibrium with a nearby susceptibility gene or polymorphism. PMID- 9352570 TI - An S-mephenytoin cysteine conjugate identified in urine of extensive but not of poor metabolizers of S-mephenytoin. AB - A conjugate of S-mephenytoin excreted in urine of extensive but not of poor metabolizers of S-mephenytoin has previously been reported. This conjugate, which is easily hydrolysed back to S-mephenytoin, has now been isolated and identified in urine from one extensive metabolizer after a single dose of 100 mg racemic mephenytoin. High performance liquid chromatography purification, followed by gas chromatographic, mass spectrometric and amino acid analyses showed that the isolated compound is a cysteine conjugate of S-mephenytoin. The significant mass spectrometric ions have been confirmed in three additional extensive metabolizers of S-mephenytoin, but were not detectable in urine from three poor metabolizer subjects. The exact structure of the conjugate is unknown, but we suggest that an S-N bond between cysteine and S-mephenytoin is formed via an oxidative radical mechanism catalyzed by CYP2C19. PMID- 9352571 TI - Genetic association between sensitivity to warfarin and expression of CYP2C9*3. AB - Cytochrome P4502C9 (CYP2C9) is largely responsible for terminating the anticoagulant effect of racemic warfarin via hydroxylation of the pharmacologically more potent S-enantiomer to inactive metabolites. Mutations in the CYP2C9 gene result in the expression of three allelic variants, CYP2C9*1, CYP2C9*2 and CYP2C9*3. Both CYP2C9*2 and CYP2C9*3 exhibit altered catalytic properties in vitro relative to the wild-type enzyme. In the present study, a patient was genotyped who had proven unusually sensitive to warfarin therapy and could tolerate no more than 0.5 mg of the racemic drug/day. PCR-amplification of exons 3 and 7 of the CYP2C9 gene, followed by restriction digest or sequence analysis, showed that this individual was homozygous for CYP2C9*3. In addition, patient plasma warfarin enantiomer ratios and urinary 7-hydroxywarfarin enantiomer ratios were determined by chiral-phase high performance liquid chromotography in order to investigate whether either parameter might be of diagnostic value in place of a genotypic test. Control patients receiving 4-8 mg warfarin/day exhibited plasma S:R ratios of 0.50 +/- 0.25:1, whereas the patient on very low-dose warfarin exhibited an S:R ratio of 3.9:1. In contrast, the urinary 7-hydroxywarfarin S:R ratio of 4:1 showed the same stereoselectivity as that reported for control patients. Therefore, expression of CYP2C9*3 is associated with diminished clearance of S-warfarin and a dangerously exacerbated therapeutic response to normal doses of the racemic drug. Analysis of the plasma S:R warfarin ratio may serve as a useful alternative test to genotyping for this genetic defect. PMID- 9352572 TI - Effects of thioridazine, an inhibitor of CYP2D6, on the steady-state plasma concentrations of the enantiomers of mianserin and its active metabolite, desmethylmianserin, in depressed Japanese patients. AB - The antidepressant mianserin is administered as a racemate of the S(+)- and R(-) enantiomers. Previous in-vitro studies have suggested that CYP2D6 is involved in the stereoselective metabolism of mianserin and its active metabolite, desmethylmianserin. To determine a role for CYP2D6 in vivo, the effects of thioridazine, an inhibitor of CYP2D6, on the steady-state plasma concentrations of the enantiomers of mianserin and desmethylmianserin were examined in 13 depressed Japanese patients. All patients were taking 30 mg of racemic mianserin at bedtime for 8-50 days. Thioridazine (40 mg/day) was coadministered for 1 week, and blood samplings were performed before and after thioridazine coadministration, 12 h after bedtime dosing. Plasma concentrations of the enantiomers of mianserin and desmethylmianserin were measured by HPLC, and the CYP2D6 genotype was determined by allele-specific PCR analysis. Thioridazine significantly increased plasma concentration of S(+)-mianserin (mean SD: 78.2 +/- 35.0 vs. 150.8 +/- 48.7 nM, P < 0.001), but not R(-)-mianserin (39.8 +/- 21.2 vs. 39.5 +/- 20.6 nM, NS). Thioridazine also significantly increased plasma concentrations of both S-desmethylmianserin (11.9 +/- 2.8 vs. 24.4 +/- 10.7 nM, P < 0.01) and R-desmethylmianserin (42.6 +/- 28.4 vs. 115.6 +/- 36.9 nM, P < 0.001). One patient homozygous for the defective allele CYP2D6*5 had the second highest and highest plasma concentrations of S(+)-mianserin and R desmethylmianserin, respectively, before thioridazine coadministration, and exhibited little increase in plasma concentration of the drugs after thioridazine coadministration. These results suggest that thioridazine specifically inhibits the metabolism of S(+)-mianserin and R-desmethylmianserin, probably through inhibition of CYP2D6, but not R(-)-mianserin. PMID- 9352573 TI - Genetically deficient CYP2D6 metabolism provides protection against oral opiate dependence. AB - Oral opiates (e.g. codeine, oxycodone, and hydrocodone) are metabolized by cytochrome CYP2D6 to metabolites of increased activity (e.g. morphine, oxymorphone and hydromorphone). CYP2D6 is genetically polymorphic, 4-10% of Caucasians lack CYP2D6 activity (poor metabolizers) due to inheritance of two non functional alleles. We tested whether the failure to activate oral opiates was a protection factor in opiate dependence by genotyping (CYP2D6*3 and CYP2D6*4 defective mutant alleles) caucasians who met or didn't meet DSM criteria for oral opiate dependence. In opiate (+/- smoking) dependent subjects we found no poor metabolizers. In contrast, the poor metabolizer frequency in never-dependent control and multi-drug dependent comparison groups was 4% and 6.5%, respectively. This under-representation of poor metabolizers (Fisher's exact test, p < or = 0.05) in people dependent on oral opiates suggests that the CYP2D6 defective genotype is a pharmacogenetic protection factor for oral opiate dependence (estimated odds ratio > 7). This is the first investigation and demonstration of differences in genetically determined P450 metabolism influencing risk for substance dependence and we suggest that these differences may influence the risk for dependence of other substrate drugs, and may occur with other genetically variable P450s. PMID- 9352574 TI - Role of CYP2D6 in the N-hydroxylation of procainamide. AB - Sequential oxidations at the arylamine moiety of the procainamide molecule leading to the formation of N-hydroxyprocainamide and its nitroso derivative may be responsible for lupus erythematosus observed in patients treated with the drug. The objective of the present study was to characterize major cytochrome P450 isozyme(s) involved in the N-hydroxylation of procainamide. Firstly, incubations were performed with microsomes from either lymphoblastoid cells or yeast transfected with cDNA encoding for specific human cytochrome P450 isozymes. Experiments performed with these enzyme expression systems indicated that the highest formation rate of N-hydroxyprocainamide was observed in the presence of CYP2D6 enriched microsomes. Additional experiments demonstrated that the formation rate of N-hydroxyprocainamide by CYP2D6 enriched microsomes was decreased from 45 +/- 4% to 93 +/- 1% by quinidine at concentrations ranging from 30 nM to 100 microM (all p < 0.05 vs control) and by approximately 75% by antibodies directed against CYP2D6. Secondly, incubations were performed with microsomes prepared from 15 human liver samples. Using this approach, an excellent correlation was observed between the formation rate of N hydroxyprocainamide and dextromethorphan O-demethylase activity (CYP2D6; r = 0.9305; p < 0.0001). In contrast, no correlation could be established between N hydroxyprocainamide formation rate and caffeine N3-demethylase (CYP1A2), coumarin 7-hydroxylase (CYP2A6), S-mephenytoin N-demethylase (CYP2B6), tolbutamide methlhydroxylase (CYP2C9), S-mephenytoin 4'-hydroxylase (CYP2C19), chlorzoxazone 6-hydroxylase (CYP2E1), dextromethorphan N-demethylase (CYP3A4), testosterone 6 beta-hydroxylase (CYP3A4/5) or lauric acid 12-hydroxylase (CYP4A11) activities. Furthermore, formation rate of N-hydroxyprocainamide was decreased in a concentration-dependent manner by quinidine (300 nM to 100 microM) and by antibodies directed against CYP2D6 but not by furafylline 20 microM (CYP1A2), ketoconazole 1 microM (CYP3A4), sulfaphenazole 10 microM (CYP2C9) or antibodies directed against CYP1A1/1A2, CYP2C, CYP2A6, CYP2E1 or CYP3A4/3A5. In conclusion, the results obtained in the present study demonstrate that CYP2D6 is the major human cytochrome P450 isozyme involved in the formation of the reactive metabolite of procainamide, namely N-hydroxyprocainamide. PMID- 9352575 TI - Specific CYP3A4 inhibitors in grapefruit juice: furocoumarin dimers as components of drug interaction. AB - Four components were isolated from grapefruit juice that inhibit human CYP3A mediated drug oxidation. The structures of these compounds were identified as furocoumarin derivatives by absorption spectra, APCI-liquid chromatography/tandem mass spectrometry and nuclear magnetic resonance after their purification by reversed-phase high performance liquid chromatography. They include two new furocoumarins, 4-[[6-hydroxy-7-[[1-[(1-hydroxy-1-methyl)ethyl]-4-methyl-6- (7-oxo 7H-furo[3,2-g][1]benzopyran-4-yl)-4-hexenyl]oxy]-3,7-dimeth yl- 2-octenyl] oxy] 7H-furo[3,2-g][1]benzopyran-7-one (GF-I-1) and 4-[[6-hydroxy-7-[[4-methyl-I- (1 methylethenyl)-6-(7-oxo-7H-furo[3,2-g][1]benzopyran-4-yl)-4- hexenyl] oxy]-3,7 dimethyl-2-octenyl]oxy]-7H-furo[3,2-g][1]benzopyran-7-one (GF-I-4). These furocoumarins are strong candidates for causative agents of grapefruit juice mediated drug interaction, because of an inhibition potential that is equal to or stronger than the prototypical CYP3A4 inhibitor, ketoconazole, on liver microsomal testosterone 6 beta-hydroxylation. PMID- 9352576 TI - The N-acetyltransferase G191 A mutation among Sudanese and Somalis. PMID- 9352577 TI - Lung cancer risk in relation to the CYP2C9 genetic polymorphism among Caucasians in Los Angeles County. PMID- 9352578 TI - Genetic analysis of CYP2C9 polymorphism in a Japanese population. PMID- 9352579 TI - CYP2D6 genotype and smoking behaviour in cigarette smokers. PMID- 9352580 TI - Effects of the CYP2D6 genotype on the steady-state plasma concentrations of haloperidol and reduced haloperidol in Japanese schizophrenic patients. PMID- 9352581 TI - Arginine-cysteine polymorphism at codon 264 of the human CYP19 gene does not affect aromatase activity. PMID- 9352582 TI - The genetic polymorphism of angiotensin-converting enzyme in a Korean population. PMID- 9352583 TI - Analgesic response in offspring of crosses between heroin delta (Swiss Webster) and mu (ICR) responding mice. PMID- 9352584 TI - Cytokines as targets for the inhibition of eosinophilic inflammation. AB - Eosinophilic inflammation is thought to play a central role in the pathogenesis of asthma. The immunoregulatory effects of interleukin (IL)-4, IL-5 and immunoglobulin (Ig)E suggest that these molecules play key roles in the effector function of eosinophils and mast cells. IL-4 regulates the development of CD4+ TH2-type cells, which elicit essential signals through IL-4 and IL-5 for the regulation of IgE production and eosinophilia, respectively. IL-5-regulated pulmonary eosinophilia and airways dysfunction can also occur independently of IL 4 and allergen-specific Igs. Such IL-4-independent pathways may also play a substantive role in the aetiology of asthma. Thus, evidence is now emerging that allergic airways disease is regulated by humoral and cell-mediated components. The essential and specific role of IL-5 in regulating eosinophilia, and the subsequent involvement of this leukocyte in the induction of lung damage and airways dysfunction, identifies IL-5 as a primary therapeutic target for the relief of airways dysfunction in asthma. PMID- 9352585 TI - Understanding and manipulating O6-methylguanine-DNA methyltransferase expression. AB - O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein that transfers methyl and alkyl lesions from the O6 position of guanine to a cysteine in its structure. The ability of MGMT to also remove precytotoxic O6-alkylguanine lesions induced by chemotherapeutic chloroethylnitrosoureas has made down regulation of MGMT expression the key component in strategies designed to sensitize tumors to the cytotoxic potential of chloroethylnitrosoureas. The study of how to regulate MGMT expression at the gene, mRNA, and protein levels has contributed not only to the development of effective inhibitors of MGMT action, but also, in a broader sense, to a better understanding of gene regulation and protein structure/function. PMID- 9352586 TI - The olfactory bulbectomized rat as a model of depression: an update. AB - The olfactory bulbectomized (OB) rat has been proposed as an animal model of depression. The following behavioural changes have been observed following bilateral olfactory bulbectomy: hyperactivity in an enclosed arena, such as the open-field; enhanced nocturnal hyperactivity in a 24-hr home cage activity monitor; deficits in memory, as shown by passive avoidance behaviour and in the Morris maze and the 8-arm radial maze; increased open arm entries in the elevated plus-maze; and changes in food motivated and conditioned taste aversion behaviour. Alterations in the noradrenergic, serotonergic, cholinergic, gamma aminobutyric acid (GABA)ergic and glutamatergic neurotransmitter systems are also associated with olfactory bulbectomy. The variety of immune changes following olfactory bulbectomy includes reduced neutrophil phagocytosis, lymphocyte mitogenesis, lymphocyte number and negative acute phase proteins, increased leucocyte adhesiveness/aggregation, monocyte phagocytosis, neutrophil number and positive acute phase proteins. An enhanced nocturnal secretion of corticosterone is observed in OB rats, which is normally suppressed by dexamethasone. The most commonly employed behavioural indicator of antidepressant activity is attenuation of the OB-related hyperactivity in the open-field. However, many of the other behavioural, neurotransmitter and immune changes have been shown to be attenuated by chronic (but not acute) antidepressant treatment. Tricyclic antidepressants (amitriptyline, desipramine), atypical agents (mianserin), selective serotonin reuptake inhibitors (paroxetine, sertraline, fluvoxamine), reversible inhibitors of monoamine oxidase A (moclobemide), as well as putative antidepressants such as 5-hydroxytryptamine1A agonists (zalospirone, ipsapirone), noncompetitive N-methyl D-aspartate antagonists (MK-801) and triazolobenzodiazepines (alprazolam, adinazolam), have demonstrated antidepressant-like activity in this model. As many of the changes exhibited by the OB rat are qualitatively similar to those observed in depressed patients, it may be concluded that the OB rat is a model of depression and not just a means whereby putative antidepressants may be tested. PMID- 9352587 TI - Oligonucleotides as modulators of cancer gene expression. AB - The delineation of gene function has always been an intensive subject of investigations. Recent advances in the synthesis and chemistry of oligonucleotides have now made these molecules important tools to study and identify gene function and regulation. Modulation of gene expression using oligonucleotides has been targeted at different levels of the cellular machinery. Triplex forming oligonucleotides, as well as peptide nucleic acids, have been used to inhibit gene expression at the level of transcription; after binding of these specific oligonucleotides, conformational change of the DNA's helical structure prevents any further DNA/protein interactions necessary for efficient transcription. Gene regulation can also be achieved by targeting the translation of mRNAs. Antisense oligonucleotides have been used to down-regulate mRNA expression by annealing to specific and determined region of an mRNA, thus inhibiting its translation by the cellular machinery. The exact mechanism of this type of inhibition is still under intense investigation and is thought to be related to the activation of RNase H, a ribonuclease that is widely available that can cleave the RNA/DNA duplex, thus making it inactive. Another well characterized means of interfering with the translation of mRNAs is the use of ribozymes. Ribozymes are small catalytic RNAs that possess both site specificity and cleavage capability for an mRNA substrate, inhibiting any further protein formation. This review describes how these different oligonucleotides can be used to define gene function and discusses in detail their chemical structure, mechanism of action, advantages and disadvantages, and their applications. PMID- 9352588 TI - Modulation of acetylcholine release in human cortical slices: possible implications for Alzheimer's disease. AB - Superfused slices of human neocortex, prepared from surgically removed tissue (to gain access to subcortical tumors) and prelabelled with [3H]choline, were stimulated electrically to evoke action potential-induced, exocytotic [3H]acetylcholine release. For comparison, rat cortex slices were also used. [3H]ACh release decreased with the age of the patients and was modulated by muscarinic autoreceptors and by 5-hydroxytryptamine1F, neurokinin1, and kappa opioid receptors located on cholinergic terminals. In addition, 5 hydroxytryptamine2 and delta-opioid receptors located on interneurons were also involved in the modulation of [3H]ACh release. The present findings might help to explain pathological conditions in Alzheimer's disease. PMID- 9352589 TI - An unusual thoracic opacity. Gossypiboma (retained surgical foreign body). PMID- 9352590 TI - The NOD mouse. PMID- 9352591 TI - Questions about NOD mouse diabetes. PMID- 9352592 TI - Insights into the chemistry and biology of the I-Ag7 class II molecule. PMID- 9352593 TI - Mechanisms underlying the loss of self tolerance in NOD mice. PMID- 9352594 TI - The role of TNF alpha and related cytokines in the development and function of the autoreactive T-cell repertoire. PMID- 9352595 TI - Immunobiology of autoimmune diabetes. PMID- 9352596 TI - CD4+ and CD8+ T lymphocytes: clarification of their pathogenic roles in diabetes in the NOD mouse. PMID- 9352597 TI - The role of CD4 and CD8 T cells in type I diabetes in the NOD mouse. PMID- 9352598 TI - Autoimmune diabetes: how many steps for one disease? PMID- 9352600 TI - Role of T-cell anergy and suppression in susceptibility to IDDM. PMID- 9352599 TI - Characterization of immunodominant peptide determinants of IDDM-associated autoantigens in the NOD mouse. PMID- 9352601 TI - Xenobiotic immunosuppressive agents: therapeutic effects in animal models of autoimmune diseases. AB - An unprecedented arsenal of new xenobiotic immunosuppressive agents has been developed recently. Most of the new immunosuppressants have been tested primarily in the treatment of allograft rejection in experimental models of transplantation, and some of the new drugs have already proven their safety and efficiency in extensive clinical trials on transplant patients. Another field for their potential application is the treatment of autoimmune diseases. This review will give an overview of the therapeutic potential of the new xenobiotic drugs in different animal models of rheumatoid arthritis, systemic lupus erythematosus, myasthenia gravis, multiple sclerosis, diabetes mellitus, thyroiditis and uveoretinitis. The new xenobiotics are either inhibitors of the de novo synthesis of nucleotides, for example mycophenolate mofetil, mizoribine, leflunomide, and brequinar, or are immunophilin-binding agents (cyclosporin, FK506 and rapamycin) that inhibit signal transduction and cell cycle progression in lymphocytes. A different mode of action is likely to account for the immunosuppressive effects of deoxyspergualin, which may interfere with intracellular chaperoning by the heat shock protein HSP70 and the activation of transcription factor NF-kappa B. PMID- 9352602 TI - Animal models of autoimmune diseases. AB - Failure of distinction between self and non-self is regarded a critical event in the pathogenesis of several human diseases such as systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, myasthenia gravis, uveoretinitis or diabetes mellitus. Autoagressive immune reactions driven by activated autoreactive lymphocytes are a characteristic feature of these autoimmune diseases. The mechanisms by which the pathogenic control of autoreactive lymphocytes deviates from physiology can be studied in appropriate animal models under well-defined experimental conditions. Experimental models of autoimmune diseases in rodent inbred strains allow for the genetic mapping of susceptibility loci and might help to identify candidate genes also relevant to the pathogenesis of human diseases. Finally, the experimental models are valuable tools to develop rational immunotherapeutic strategies. Interesting features of some of the models employed for such research will be introduced in this review. PMID- 9352603 TI - Outcome of polyarteritis nodosa in northern India. AB - Over the last 10 years, 17 patients (13 males and 4 females) diagnosed as having classical polyarteritis nodosa (PAN) were treated and followed up in the rheumatology clinic of our institute. The median age and duration of symptoms at presentation were 29 years (range 13-59) and 9.5 months, respectively. Patients presented with the typical clinical picture of classical PAN. The diagnosis was established with the help of an aortogram (7/10), sural nerve biopsy (7/8), muscle biopsy (5/7) and renal biopsy (3/4). For reasons not known, none of the 17 patients was HBsAg positive. Patients were treated with a combination of oral prednisolone (1 mg/kg per day) for 6 weeks, which was slowly tapered off over 6 months, and monthly intravenous cyclophosphamide pulses (15 mg/kg) for the first 6 months, followed by 3-monthly pulses for a total of 2 years. Remission was achieved in 14 patients after a median of 5 months of treatment. Remission was stable for a median of 5 years of follow-up. Three patients did not respond well and died within 6 months of diagnosis. The causes of death in these were intracerebral haemorrhage in one patient and gastrointestinal bleeding in two patients. This experience is in accord with the reported literature that classical PAN is mostly a monophasic disease with either an excellent response to the appropriate immunosuppressive therapy and a long remission or a downhill course culminating in death. A chronic course is rare. PMID- 9352604 TI - Radiosynoviorthesis with rhenium-186 in rheumatoid arthritis: a prospective study of three treatment regimens. AB - The aim of this study was to evaluate the efficiency of radiation synovectomy with rhenium-186 in rheumatoid arthritis. In this prospective, randomized trial we compared three different treatment regimens for shoulder, elbow, wrist, hip and ankle joints: group 1, injection of rhenium-186; group 2, injection of rhenium-186 in combination with triamcinolone hexacetonide; group 3, injection of triamcinolone hexacetonide alone. Each treatment group included 50 joints. Patients included in the study had to fulfil the following criteria: (1) they had to have a diagnosis of rheumatoid arthritis (ARA criteria 1988), (2) their disease-modifying drug had to be methotrexate, started at least 6 months prior to injection therapy and given for the entire study time, (3) their nonsteroidal anti-inflammatory drug had to be diclofenac given at a dose of 150 mg/day or less and (4) they were also given prednisolone at a dose of 7.5 mg/day or less. After 3 years of follow-up, 79 joints met these criteria, i.e. 71 joints were excluded from the study: 26 joints because the patients changed the disease-modifying drug (12 joints from group 1, 4 joints from group 2 and 10 joints from group 3); 45 joints because of recurrent synovitis and second-stage treatment (21 joints from group 1, 5 joints from group 2 and 19 joints from group 3). During the follow-up period, joints were assessed for pain, synovitis, joint motion and stage of radiological destruction. Best clinical results and slowest progression in radiological destruction were achieved with the combined injection of rhenium-186 and triamcinolone hexacetonide. Therefore, we recommend this treatment for articulosynovitis with the exception of severe forms, the latter because of the effective penetration range of rhenium-186. PMID- 9352605 TI - Measurement of transcription factor c-fos and EGR-1 mRNA transcription levels in synovial tissue by quantitative RT-PCR. AB - The transcription factors Fos and EGR-1 are known to be involved in the regulation of the transcription of metalloproteinases and their specific inhibitors. Since the overexpression of metalloproteinases is responsible for the matrix degradation in rheumatoid arthritis (RA), exact analysis of transcription levels of c-fos and EGR-1 ex vivo may serve to monitor progression or remission of the disease activity in RA. Here we report on a method based on a quantitative reverse transcription polymerase chain reaction (RT-PCR) for rapid estimation of the transcription levels of the immediate early genes c-fos and EGR-1. Coamplification with suitable internal standards, easily generated by the use of hybrid primers, allows us to semiquantitatively measure c-fos and EGR-1 induction levels in low numbers of cultured cells or very small tissue samples obtained by synovial biopsy. The sensitivity of the method was 3.5 pg/ml for c-fos- and 10 pg/ml for EGR-1-specific cDNAs. PMID- 9352606 TI - Immuno-localisation of tumour necrosis factor and its receptors in temporal arteritis. AB - Temporal arteritis (TA) is an acute vasculitis characterised by destruction of arterial architecture following infiltration of the arterial wall by macrophages, giant cells and lymphocytes. Using immunohistochemical techniques, tumour necrosis factor (TNF) was demonstrated in up to 60% of the cells in all areas of inflamed arteries. More cells staining for TNF were detected in the intima and media of inflamed vessels than control uninflamed arteries (P < 0.003 and P < 0.001, respectively). In TA, TNF was localised to giant cells and macrophages, suggesting that its predominant source is from the monocyte lineage, but, occasionally, TNF staining was found in areas infiltrated by T cells. Many endothelial cells also contained TNF, but there were no differences between the number of endothelial cells staining in inflamed and normal blood vessels. Of the two TNF receptors, the p75 receptor was sparsely represented in the inflamed vessels in TA. By comparison, the p55 receptor was widely detected on endothelial cells and infiltrating mononuclear cells close to the internal elastic lamina (IEL). Endothelial cells from normal vessels also stained for both TNF receptors, but normal smooth muscle cells in the vessel media expressed the p55 receptor, indicating that they are capable of responding to locally secreted TNF. Localisation of TNF receptors and TNF in close proximity to the IEL suggests that TNF could be involved in the leucocyte infiltration and arterial wall destruction characteristic of TA. PMID- 9352607 TI - Assessing health in musculoskeletal disorders--the appropriateness of a German version of the Sickness Impact Profile. AB - The Sickness Impact Profile (SIP) is gaining increasing popularity in clinical and epidemiological studies, assessing health status or the impact of therapeutic interventions. The SIP was created as a global measure, assessing generic dimensions of health. As there were only a few such methods available in our country, we decided in 1990 to validate a German version of the SIP and to test its appropriateness in patients with musculoskeletal disorders. Due to the up-to dateness of our results, we decided to publish now the data of this German version of the SIP. Reliability, validity and sensitivity to change of the German version of the SIP were tested in 299 patients with musculoskeletal disorders. The test-retest correlation was high for the overall score (r = 0.81), as well as for Cronbach's alpha (r = 0.83). Results for categories were significantly lower. Validity was tested by comparing the SIP to the Keitel Index (r = 0.6) and to the Measurement of Patient Outcome Scale (r = 0.72), as well as to a control group, which showed significant differences in all categories and in the overall score. Furthermore, the SIP was able to demonstrate therapy-related improvements in health in all our patients. We concluded that the german version of the SIP fulfilled statistical test criteria. The results were comparable to those achieved in the original American version. A cross-cultural comparison using the SIP is justified for at least those patients with musculoskeletal disorders. PMID- 9352608 TI - A fatal case of severe SLE complicated by invasive aspergillosis. AB - We report on the case of a 25-year-old female with severe systemic lupus erythematosus (SLE) who presented with pancytopenia, fever, arthralgia and abdominal pain. After antibiotic treatment, the patient was afebrile for 3 days before her temperature rose again. Dyspnoea and cough pointed towards pneumonia which was confirmed by X-ray. Different antibiotics and the antimycotic agent fluconazol were given. The lupus flare was treated with high-dose prednisolone. After a couple of days, the dyspnoea increased and mechanical ventilation became necessary. Bronchoscopy and transbronchial biopsy revealed the diagnosis of invasive aspergilloses. Despite of an immediate treatment with amphotericin B, the patient died because of respiratory insufficiency. The literature on aspergillosis in SLE is reviewed and prophylactic, diagnostic and therapeutic options are discussed for this infectious complication which has an 80% mortality in patients with SLE. PMID- 9352609 TI - The interplay of nitric oxide and peroxynitrite with signal transduction pathways: implications for disease. AB - Since the discovery that at least one form of endothelium derived relaxing factor is nitric oxide (NO), numerous studies have uncovered diverse roles for this free radical in a variety of physiological and pathophysiological processes. NO production, a process mediated by a family of enzymes termed NO synthases, has been detected in most cell types. Many of the effects of NO are thought to be mediated through its direct interaction with specific and defined cell signaling pathways. The nature of such interactions are highly dependent on the concentration of NO and cell type. Furthermore, specific NO derived reaction products, such as peroxynitrite, also have the potential to effect cell signal transduction events. As with NO, this can occur through diverse mechanisms and depends on concentration and cell type. It is perhaps not surprising that the reported effects of NO in different disease states are often conflicting. In this brief overview, a framework for placing these apparently disparate properties of NO will be described and will focus on the effects of NO and peroxynitrite on signaling pathways. PMID- 9352610 TI - Nitric oxide and pregnancy. AB - We have hypothesized that an alteration in the production of endothelium dependent factors by sex hormones is a potential unifying mechanism for both the decreased arterial contractility and the redistribution of cardiac output characteristic of normal pregnancy. Thus, the effect of pregnancy/ estradiol on any one vascular bed will reflect the number and distribution of estrogen receptors. In this article, we review what is known about the effects of pregnancy and estrogen on nitric oxide synthase. Pregnancy increases Ca(2+) dependent NOS activity early in gestation. The timing of the increase parallels the increase in plasma estradiol concentration. The increase in maternal brain NOS during pregnancy is blocked by tamoxifen. cGMP content increases along a similar time course in most but not all tissues. The changes in cGMP more closely approximate the changes in blood flow during pregnancy. This suggests that multiple elements of the NO:cGMP pathway are altered by pregnancy. It also shows that cGMP content cannot always be used as a surrogate for NOS activity. Estradiol, but not progesterone or testosterone, increases CA(2+)-dependent NOS activity. NO accounts for some, but not all of the pregnancy-associated changes in maternal arterial contractile response. It is not involved in uterine quiescence. Nitric oxide synthase is developmentally regulated in the fetus and is likely important in regulating the distribution of fetal blood flow. PMID- 9352611 TI - Ontogeny of nitric oxide in the pulmonary vasculature. AB - The vasodilator molecule nitric oxide is critically involved in the successful cardiopulmonary transition from fetal to postnatal life. It is produced in the pulmonary endothelium by the endothelial isoform of the enzyme nitric oxide synthase. The expression of endothelial nitric oxide synthase in the lung increases dramatically during late gestation, optimizing the capacity for nitric oxide production at the time of birth. Studies in cultured cell models indicate that the developmental upregulation may be mediated by estrogen, and that the expression of the enzyme is also upregulated by oxygen. Pulmonary endothelial nitric oxide synthase expression is diminished in models of congenital diaphragmatic hernia and neonatal pulmonary hypertension induced by fetal ductal ligation. Thus, there is normally a marked developmental upregulation in endothelial nitric oxide synthase expression in the lung during late fetal life, and attenuated expression of the enzyme may contribute to the pathophysiology of a variety of forms of neonatal pulmonary vascular disease. PMID- 9352612 TI - Models of persistent pulmonary hypertension of the newborn (PPHN) and the role of cyclic guanosine monophosphate (GMP) in pulmonary vasorelaxation. AB - At birth, a marked decrease in pulmonary vascular resistance allows the lung to establish gas exchange. Persistent pulmonary hypertension of the newborn (PPHN) occurs when this normal adaptation of gas exchange does not occur. We review animal models used to study the pathogenesis and treatment of PPHN. Both acute models, such as acute hypoxia and infusion of vasoconstrictors, and chronic models of PPHN created both before and immediately after birth are described. Inhaled nitric oxide is an important emerging therapy for PPHN. We review nitric oxide receptor mechanisms, including soluble guanylate cyclase, which produces cGMP when stimulated by nitric oxide, and phosphodiesterases, which control the intensity and duration of cGMP signal transduction. A better understanding of these mechanisms of regulation of vascular tone may lead to safer use of nitric oxide and improved clinical outcomes. PMID- 9352613 TI - Nitric oxide and platelet function: implications for neonatology. AB - Nitric oxide (NO) is a mediator that modulates vessel wall tone and hemostatic thrombotic balance. Platelet function is regulated by NO generated from platelets, endothelial cells and leukocytes. Nitric oxide has been shown to inhibit platelet adhesion, aggregation, and stimulate disaggregation of preformed platelet aggregates. Many of the effects of NO are mediated by its stimulation of guanylate cyclase and the formation of cyclic GMP and its subsequent transduction mechanism. In vivo, NO is likely to interact with prostacyclin, metabolites of ecto-nucleotidase, and lipoxygenase to modulate platelet function in a synergistic manner. An imbalance of NO production (deficiency or overproduction) has been implicated in the pathogenesis of various vascular disorders including thrombosis, atherosclerosis, septicemia, and ischemia-reperfusion injury. It is likely that some of detrimental effects of NO are mediated through its reaction with superoxide anion to form the potent oxidant, peroxynitrite. Nitric oxide gas and NO donors are used for the pharmacological treatment of various vascular disorders. Because inhaled NO has been documented to improve systemic oxygenation and reduce the need for extracorporeal membrane oxygenation, it has been widely used in neonates with severe hypoxemia. An inhibition of platelet function, resulting in a prolonged bleeding time, has been shown in adults receiving inhaled NO. Because bleeding complications may occur in high-risk infants, it is important to evaluate the effect of inhaled NO on platelet function and its correlation with clinical consequences such as intracranial hemorrhage. For these reasons, hemostasis should be carefully monitored during the administration of inhaled NO to critically ill neonates. PMID- 9352614 TI - Inhaled nitric oxide in the premature infant: animal models and clinical experience. AB - Inhaled nitric oxide (iNO) is an effective adjuvant therapy for term newborns with persistent pulmonary hypertension. However, its role in treating hypoxemic respiratory failure in premature newborns has not been established. Laboratory experiments have shown the importance of endogenously produced NO in fetal and neonatal pulmonary vasoregulation in the premature lamb. Moreover, low-dose iNO improves oxygenation and reduces pulmonary vascular resistance in the premature lamb with hyaline membrane disease. Preliminary studies have suggested the potential role of low-dose iNO in premature newborns with hyaline membrane disease, sepsis, and pulmonary hypoplasia. However, prematurity poses unique risks that must be carefully addressed with clinical trials designed to measure both safety and efficacy of this promising new therapy. PMID- 9352615 TI - Nitric oxide in respiratory failure in the newborn infant. AB - Nitric oxide given as an inhalation (INO) is a novel selective pulmonary vasodilator without effects on the systemic circulation. Preliminary observations indicated that INO treatment was associated with improvements in oxygenation in near-term newborn infants with hypoxic respiratory failure and persistent pulmonary hypertension of the newborn (PPHN). Subsequently, at least eight prospective randomized controlled trials evaluating the use of INO in the near term neonate with hypoxic respiratory failure have been presented or published. A meta-analysis of these trials has provided evidence that INO improves the PaO2 in the INO-treated infants by 52.8 mm Hg (weighted mean difference) compared with controls (95% CI, 38.2, 67.4), and significantly decreases the oxygenation index by 16.9 compared with controls (95% CI, -22.2, -11.6). The incidence of death or need for ECMO is significantly reduced by treatment with INO, relative risk 0.71 compared to control (95% CI, 0.57, 0.87), with the majority of the improvement observed in the reduction in the need for ECMO. A single study of infants with congenital diaphragmatic hernia (CDH) did not show a benefit for early INO therapy, with treated infants having a greater requirement for ECMO (P = .043). At present, there are no long-term evaluations of infants who have received INO as part of these prospective trials. INO improves oxygenation and reduces the need for ECMO in the near-term hypoxic neonate, but further research is required to evaluate the ultimate safety and benefit of this therapy. PMID- 9352616 TI - Inhaled nitric oxide in the neonate with cardiac disease. AB - As a selective pulmonary vasodilator, inhaled nitric oxide is an important diagnostic and therapeutic agent for the treatment of pulmonary hypertension in patients with congenital heart disease. Among 400 patients treated in our center with nitric oxide, 37% were newborns. Hemodynamic benefit was shown in newborns with total anomalous pulmonary venous connection, in those with congenital mitral stenosis, and in postoperative patients with preexisting left to right shunts and other lesions. It can be used to help discriminate anatomic obstruction to pulmonary blood flow from pulmonary vasoconstriction, and it may be used in the treatment or prevention of pulmonary hypertensive crises after cardiopulmonary bypass. However, none of the purported benefits of inhaled nitric oxide in children with congenital heart disease have been studied in a randomized, placebo controlled manner. PMID- 9352617 TI - On surgical intervention in the temporomandibular joint. AB - The aim of this thesis was to evaluate the indications for and the results of temporomandibular joint (TMJ) surgery in patients with long-standing severe orofacial pain and dysfunction as well as in patients with fractures of the condylar neck. The patients with long-standing pain and dysfunction had had symptoms for a mean time of 4 years, had been treated conservatively for a mean time of 2.5 years, and had undergone numerous conservative treatment methods without improvement except for a minor increase in mouth opening capacity. The indications for surgery were strict; only 1% or less of all the patients referred to the departments with a diagnosis of temporomandibular disorder (TMD) were prescribed surgery, which was considered to be the only remaining option. The TMJ surgery reduced pain, sleeping problems, and analgesic consumption and improved mouth opening capacity. The procedure showed low morbidity except for a facial nerve disturbance in three patients. Postoperatively, the bite force was observed to be normalised, and the radiographic examination showed moderate to severe osteoarthrotic changes. These changes, though extensive, were considered to be the normal outcome of diskectomy and without clinical significance, even though they resembled degenerative joint disease. In study V, surgery was performed on patients with a clear diagnosis of anterior disk displacement (ADD) with or without reduction. The preoperative pain and mouth opening capacity were markedly improved as well as other subjective symptoms. Although surgical morbidity was low, some radiographic changes were clearly detectable. In agreement with earlier reports, patients with a distinct diagnosis of ADD with or without reduction were clearly helped by diskectomy. In cases of ADD with or without reduction, it can be concluded that unsuccessful conservative treatment should not exceed 3-6 months but be discontinued in favour of the documented advantages of surgery in these cases. Patients with ankylosis should be treated surgically without delay. Unclear diagnoses such as arthralgia and osteoarthrosis with symptoms should be excluded from surgery unless overlapping muscular hyperactivity has been excluded as a major cause of the patients problem. Diskectomy is a useful surgical procedure for patients with severe long-standing TMD. It was shown in study VI that patients with dislocated fractures of the condylar neck can be successfully treated with open surgical reduction when the dislocation is large and associated with symptoms and limited function. When cognitive-behavioural profiles were measured psychometrically in study VII, a dysfunctional profile was more common in patients with myofascial pain and pain with an obscure origin than in other patients diagnosed with TMD. The dysfunctional profile was also common in patients in whom treatment of a conservative or surgical nature had failed. Among TMD patients with disk displacement, adaptive copers were most common in successfully diskectomized patients and least common in patients about to undergo invasive treatment. PMID- 9352618 TI - Studies of occlusal adjustment therapy in patients with craniomandibular disorders. PMID- 9352620 TI - Ultrasonic power Doppler imaging for prostatic cancer: a preliminary report. AB - This study was conducted to characterize Doppler blood flow signals in prostatic cancer using power Doppler imaging with a transrectal probe. Both the localization and vascularity of blood flow in the prostate were compared between patients with prostatic cancer and those with benign prostatic hyperplasia (BPH). The prominent accumulation of blood flow signals (hypervascular lesion) was recognized in all the cases with prostatic cancer, compared to only 2 (4%) of 47 with BPH (p < 0.001). Power Doppler imaging is promising for the detection of prostatic cancer. PMID- 9352619 TI - Why is the hypotensive effect of clonidine greater in hypertensive rats? AB - The original aim of this study was to observe whether the depressor drug clonidine inhibited the abnormal hindquarter tone in spontaneously hypertensive rats (SHR). In conscious SHR and normotensive control rats (NCR), hindquarter (terminal aortic) blood flow was observed with an implanted electromagnetic flow probe and mean arterial pressure with an indwelling catheter. Twenty minutes after intravenous injection of clonidine (5 micrograms/kg) when arterial pressure reached a steady lower level, hindquarter resistance (HQR), calculated as mean arterial pressure divided by hindquarter flow, did not decrease in SHR. Thus we were unable to obtain evidence for an inhibitory effect of clonidine on the abnormal hindquarter tone in SHR. In NCR, HQR increased significantly by clonidine. The decrease in arterial pressure on clonidine was greater in SHR than in NCR, presumably because the increase in HQR partially offset the hypotensive effect in NCR. It seems that the increase in HQR in NCR was induced by a reflexive excitation of regional sympathetic vasoconstrictor fibers, which, being the final common path for the abnormal hindquarter tone also, were already being excited in SHR before clonidine administration. This point was quantitatively verified. PMID- 9352622 TI - Central and peripheral effects of the non-neural substances on respiration before and after vagotomy. AB - The central effects of capsaicin, veratrine, histamine and bradykinin were studied by injecting them directly into the oerebrospinal fluid and their peripheral effects were examined by injecting into femoral vein. Our experiments were performed in Na-pentobarbital-anaesthetized dogs. Tidal volume (VT), respiratory frequency (f/min), systemic arterial pressure (BP) were recorded. A significant increase in f, and an initial apnea or hypoventilation followed by a significant increase in VT were observed with central and peripheral capsaicin. Vagotomy removed the peripheral VT response, but not the central one. While central capsaicin administration increased BP, peripheral administration decreased. After vagotomy, a significant increase was observed in BP for both administrations. Respiratory responses to central and peripheral administrations of veratrine were similar to those of capsaicin. Significant increases were observed in f and VT of the intact group in response to central and peripheral administration of histamine. Response to peripheral administration disappeared after vagotomy. While central and peripheral bradykinin increased VT significantly, there was no significant change in f. Vagotomy only removed the increase in VT in response to peripheral administration. In conclusion, respiratory responses to central administration of capsaicin and veratrine are due to direct effects of these substances on respiratory neurons. In peripheral administration, disappearance of the responses after vagotomy indicate that the responses are brought about by stimulation of the lung receptors. PMID- 9352623 TI - Genetically determined thymus enlargement in the early life in BUF/Mna rats. AB - BUF/Mna (B) rat is a mutated strain, having much larger thymus than WKY/ NCrj (W), ACI/NMs (A), and F344 (F) rats throughout their life-span. Rats of the latter 3 strains have normal sized thymuses, being less than 5.3 in the thymus to body weight ratio (mg/g), when they were killed at 6 weeks of age. Genetic segregation of large thymus size in the B strain at 6 weeks of age was studied by crossing B rats with W, A or F rats. All of 3 types of the F1 hybrid rats between the B strain and the other strains showed intermediate thymus ratios between those of both parental strains. In F2 rats between the B and W strains, the distribution of thymus ratios showed about 5 different peaks. These findings might indicate that two polymeric autosomal loci, thymus enlargement-1 (Ten-1) and thymus enlargement-2 (Ten-2), can enlarge the thymus size in B rats. Histometrically, whole thymus and cortex areas of the B rats were 2-5 times larger than the W rats during 6-12 weeks of age, but medulla areas were slightly different between the strains, showing that larger thymuses in B rats were mainly due to the enlarged cortex areas. PMID- 9352621 TI - Roles of the visceral pleura in the production of pleural effusion in permeability pulmonary edema. AB - We investigated the roles of the mesothelium of the visceral pleura on hydraulic conductivity in dogs under normal conditions and condition of permeability pulmonary edema. Nineteen mongrel dogs were divided into following 4 groups: thoracotomy alone (control group, n = 7); thoracotomy and striping of the mesothelium using Gelfilm (C + G group, n = 4); injection of oleic acid to increase the permeability of the pulmonary vessels (OA group, n = 4); injection of oleic acid and striping of the mesothelium (OA + G group, n = 4). A hemispherical capsule filled with physiological saline was attached to the visceral pleura. The transpleural fluid flow (delta V) was measured at given incremental or decremental hydrostatic pressures (delta Pcap) in the capsule. Hydraulic conductivity was calculated from the slope of linear regression line obtained from relationship between delta Pcap and the fluid flow rate (v) according to the Starling's equation. The conductivity obtained were 1.49 +/- 0.69 (nl.min-1.cmH2O-1.cm-2) in the control group, 1.37 +/- 0.88 in the C + G group, 3.75 +/- 0.74 in the OA + G group, and 7.07 +/- 2.49 in the OA + G group. The hydraulic conductivity was not increased by striping of the mesothelium (1.49 +/- 0.69 [nl.min-1.cmH2O-1.cm-2] vs. 1.37 +/- 0.88, in the control group vs. C + G group, respectively). Visceral pleural hydraulic conductivity following OA injection was increased by striping of the mesothelium (3.75 +/- 0.74 vs. 7.07 +/ 2.49 in OA group vs. OA + G group, respectively). These findings suggest that the wall of pulmonary vessels acts as a barrier to movement of pleural effusion under normal conditions, whereas the mesothelium of the visceral pleura acts as that under condition of permeability pulmonary edema. PMID- 9352624 TI - Serum secretory leukoprotease inhibitor levels in chronic bronchitis and Sjogren's syndrome. AB - Serum secretory leukoprotease inhibitor (SLPI) is synthesized and secreted by serous cells in airway glands, and the serum level is speculated to reflect airway gland hyperplasia. To test this hypothesis, we measured the serum SLPI in 38 clinically stable patients with chronic bronchitis (3F and 35M; 58 +/- 2 years, mean +/- S.E.M.) (group CB), 24 patients with Sjogren's syndrome (24F; 52 +/- 3 years) (group SG), and compared it with 12 healthy control subjects (6F and 6M; 54 +/- 3 years) (group CN) using an enzyme-linked immunosorbent assay (ELISA). The serum SLPI from group CB (105 +/- 8 ng/ml) was significantly higher than that from group CN (60 +/- 2 ng/ml) and further, it significantly correlated with sputum volume per day. Although the mean value of serum SLPI from group SG (64 +/- 5 ng/ml) was not different from that from group CN, serum SLPI significantly correlated with the duration of respiratory symptoms (cough and/or sputum) in group SG. In conclusion, serum SPLI level reflects airway gland hyperplasia, suggesting that SLPI measurement is a possible laboratory method to estimate airway glandular hyperplasia. PMID- 9352626 TI - Changes in left ventricular diastolic filling patterns before and after the closure of the ductus arteriosus in very-low-birth weight infants. AB - To evaluate serial changes in left ventricular diastolic filling patterns in preterm infants, we performed echocardiographic examinations in 18 very-low-birth weight infants and 20 fullterm infants before and after the closure of the ductus arteriosus. In the fullterm infants, the ductal closure induced significant decreases in the peak velocity and flow velocity integral of early diastole, first third filling fraction, and mitral stroke volume. In the preterm infants, by contrast, there were significant increases in the flow velocity integral of early diastole, first third filling fraction, and mitral stroke volume after the ductal closure. No differences following the ductal closure were found in the atrial phase of filling and peak filling rate normalized to stroke volume in either group. When the ductus arteriosus was open, essentially the same left-to right shunting of the ductus arteriosus was detected in both preterm and fullterm infants, but the Doppler flow patterns of the patent foramen ovale were different: the fullterm infants had a single flow peak mainly during ventricular late systole and early diastole, but the preterm infants had two or three flow peaks with nearly equal amplitudes lasting from ventricular systole to diastole, which resembled the Doppler flow pattern of atrial septal defect. Only a faint Doppler flow signal of the foramen ovale was observed after the ductus arteriosus closed. Our results obtained from the preterms suggest that the left-to-right shunt through the foramen ovale may be one important factor to alter the Doppler transmitral filling patterns during the fetal to neonatal cardiovascular changes. PMID- 9352625 TI - Gene expression of MASH-1, MATH-1, neuroD and NSCL-2, basic helix-loop-helix proteins, during neural differentiation in P19 embryonal carcinoma cells. AB - We examined the gene expression of MASH-1, MATH-1, neuroD and NSCL-2 during neural differentiation of P19 embryonal carcinoma cells using reverse transcription-polymerase chain reaction and high performance liquid chromatography. These proteins are members of basic helix-loop-helix transcription factor family and their expressions are reported to be transient and restricted in the nervous system during early neurogenesis. Retinoic acid (RA , 1 microM)-treatment and aggregation for 4 days induced and greatly increased MASH-1, neuroD and NSCL-2 mRNA in P19 cells. The increases peaked at day 3, 4 and 5, respectively. RA-treatment increased MATH-1 mRNA slightly. mRNA of MAP2, a neural differentiation marker, were increased by RA-treatment and the increases reached to the plateau at day 5. The results indicate that the gene expression of MASH-1, MATH-1, neuroD and NSCL-2 during neural differentiation in P19 cells is transient and the order is similar to that in the mouse embryo nervous system as previously reported. PMID- 9352627 TI - Acute mercury poisoning by intentional ingestion of mercuric chloride. AB - A 26-year-old woman ingested 0.9 g of mercuric chloride in a suicide attempt and developed hematemesis, melena and acute renal failure. Anuria persisted for 14 days. She was treated by plasma exchange, hemodialysis and peritoneal dialysis in combination with continued dimercaprol chelation. While hemodialysis was ineffective in removing the mercury, plasma exchange effectively eliminated mercury. After two plasma exchange therapies, mercury concentration in the blood decreased linearly on a log scale with half-lives of 23.1 days for whole blood and 19.1 days for plasma, using first-order kinetics. One month after ingestion, renal function recovered to normal as judged by serum creatine and blood urea nitrogen levels, although the beta 2-microglobulin level in urine was still elevated. At a follow-up examination four months later, renal function was found to be completely normal. This indicates that the renal damage caused by acute mercuric chloride poisoning may not be permanent. PMID- 9352628 TI - Boris Konstantinovich Vainshtein 1921-1996. PMID- 9352629 TI - Polytene chromosomes, heterochromatin, and position effect variegation. PMID- 9352630 TI - The role of retinal bipolar cell in early vision: an implication with analogue networks and regularization theory. AB - A linear analogue network model is proposed to describe the neuronal circuit of the outer retina consisting of cones, horizontal cells, and bipolar cells. The model reflects previous physiological findings on the spatial response properties of these neurons to dim illumination and is expressed by physiological mechanisms, i.e., membrane conductances, gap-junctional conductances, and strengths of chemical synaptic interactions. Using the model, we characterized the spatial filtering properties of the bipolar cell receptive field with the standard regularization theory, in which the early vision problems are attributed to minimization of a cost function. The cost function accompanying the present characterization is derived from the linear analogue network model, and one can gain intuitive insights on how physiological mechanisms contribute to the spatial filtering properties of the bipolar cell receptive field. We also elucidated a quantitative relation between the Laplacian of Gaussian operator and the bipolar cell receptive field. From the computational point of view, the dopaminergic modulation of the gap-junctional conductance between horizontal cells is inferred to be a suitable neural adaptation mechanism for transition between photopic and mesopic vision. PMID- 9352631 TI - A model for learning human reaching movements. AB - Reaching movement is a fast movement towards a given target. The main characteristics of such a movement are straight path and a bell-shaped speed profile. In this work a mathematical model for the control of the human arm during ballistic reaching movements is presented. The model of the arm contains a 2 degrees of freedom planar manipulator, and a Hill-type, non-linear mechanical model of six muscles. The arm model is taken from the literature with minor changes. The nervous system is modeled as an adjustable pattern generator that creates the control signals to the muscles. The control signals in this model are rectangular pulses activated at various amplitudes and timings, that are determined according to the given target. These amplitudes and timings are the parameters that should be related to each target and initial conditions in the work-space. The model of the nervous system consists of an artificial neural net that maps any given target to the parameter space of the pattern generator. In order to train this net, the nervous system model includes a sensitivity model that transforms the error from the arm end-point coordinates to the parameter coordinates. The error is assessed only at the termination of the movement from knowledge of the results. The role of the non-linearity in the muscle model and the performance of the learning scheme are analysed, illustrated in simulations and discussed. The results of the present study demonstrate the central nervous system's (CNS) ability to generate typical reaching movements with a simple feedforward controller that controls only the timing and amplitude of rectangular excitation pulses to the muscles and adjusts these parameters based on knowledge of the results. In this scheme, which is based on the adjustment of only a few parameters instead of the whole trajectory, the dimension of the control problem is reduced significantly. It is shown that the non-linear properties of the muscles are essential to achieve this simple control. This conclusion agrees with the general concept that motor control is the result of an interaction between the nervous system and the musculoskeletal dynamics. PMID- 9352632 TI - Self-organizing effects of spontaneous neural activity on the development of spinal locomotor circuits in vertebrates. AB - Presented in this paper is a neural network model that can be used to investigate the possible self-organizing mechanisms occurring during the early ontogeny of spinal neural circuits in the vertebrate motor system. The neural circuit is composed of multiple types of neurons which correspond to motorneurons, Renshaw cells and a hypothetical class of interneurons. While the connectivity of this circuit is genetically predetermined, the efficacies of these connections--the synaptic strengths--evolve in accordance with activity-dependent mechanisms which are initiated by the intrinsic, autonomous activity present in the developing spinal cord. Using Oja's rule, a modified Hebbian learning scheme for adjusting the values of the connections, the network stably self-organizes developing, in the process, reciprocally activated motorneuron pools analogous to those which exist in vivo. PMID- 9352633 TI - Learned changes in the complexity of movement organization during multijoint, standing pulls. AB - This paper tests the hypothesis that the central nervous system (CNS) learns to organize multijoint movements during a multijoint 'bouncing pull' task such that, after practice, motion of the anterior-posterior center of mass (CMAP) more closely resembles that of a conservative, one degree of freedom (DF), inverted pendulum model. The task requires standing human subjects to produce precise peak pulling forces on a handle while maintaining balance-goals that can be easily accomplished if movement is organized as in the model. Ten freely standing subjects practiced making brief, bouncing pulls in the horizontal direction to target forces (20-80% of maximum) for 5 days. Pulling force, body kinematic and force plate data were recorded. An eight-segment analysis determined sagittal plane CM motion. We compared the effects of practice on the regression-based fit between actual and model-simulated CMAP trajectories, and on measures of CMAP phase plane symmetry and parameter constancy that the model predicts. If the CNS learns to organize movements like the inverted pendulum model, then model fit should improve and all other measures should approach zero after practice. The fit between modeled and actual CMAP motion did not improve significantly with practice, except for moderate force pulls. Nor did practice increase phase plane symmetry or parameter constancy. Specifically, practice did not decrease the differences between the pre-impact and rebound positions or speeds of the CMAP, although speed difference increased with pulling force. CMAP at the end of the movement was anterior to its initial position; the anterior shift increased after practice. Differences between the pre-pull and balance-recovery ankle torque (TA) impulses were greater on day 5 and correlated with the anterior shift in CMAP. These results suggest that practice separately influenced the force production and balance recovery phases. A modified model with damping could not explain the observed behaviors. A modified model using the actual time-varying TA profiles improved fit at lower force levels, but did not explain the increased postural shift after practice. We conclude that the CNS does not learn to organize movements like the conservative, inverted pendulum model, but rather learned a more complex form of organization that capitalized on more time-varying controls and more intersegmental dynamics. We hypothesize that at least one additional DF and at least one time-varying parameter will be needed to explain fully how the CNS learns to organize multijoint, bouncing pulls made while standing. PMID- 9352634 TI - Fast cortical selection: a principle of neuronal self-organization for perception? AB - It is commonly accepted that larger visual objects are represented in the cerebral cortex by specific spatial patterns of neuronal activity. Self organization is a key concept in the different explanations of such neuronal representations. We here propose as a hypothesis that fast cortical selection (FCS) is an intrinsic functional element of cortical self-organization during perception. Selection is a central concept in theoretical biology which has proved its explanatory power in different fields of our natural and cultural world. The central element in the cortical selection process is the pyramidal cell with its two types of excitatory input. In primary cortical areas one of these inputs comes from any of the sensory organs, determining the topological and typological receptive field properties of the cell and also driving it directly. The other type of input connects reciprocally neighbouring pyramidal cells by axon collaterals and only facilitates the driving input. These two functionally different inputs constitute the elementary selection system working by iterative mutual facilitation as a biological algorithm. A short simulation, based entirely on such biological facts, illustrates the dynamic of this selection process: the activity of cells responding better to the external stimulus 'grow and survive' the stimulation, whereas less responsive cells decrease their activity due to competition. PMID- 9352635 TI - Action patterns and mapping of the substrate-binding regions of endo-(1-->5) alpha-L-arabinanases from Aspergillus niger and Aspergillus aculeatus. AB - The substrate binding sites of endo-(1-->5)-alpha-L-arabinanases (EC 3.2.1.99) from Aspergillus niger and Aspergillus aculeatus were investigated using reduced and regular (1-->5)-alpha-L-arabino-oligosaccharides and high performance anion exchange chromatographic analysis. Calculation of bond cleavage frequencies and kcat/K(m) parameters for these substrates enabled the determination of the number of arabinofuranosyl binding subsites and the estimation of the binding affinities of each subsite. The A. aculeatus endo-arabinanase has six subsites arranged symmetrically around the catalytic site, while the A. niger endo-arabinanase has five subsites; two from the catalytic site towards the non-reducing end of the bound substrate and three toward the reducing end. The two subsites directly adjacent to the catalytic sites in both the A. niger and A. aculeatus endo arabinanase have near-zero net free energy of binding. These results are unlike most glycopyranosyl endo-hydrolases studied which have net negative (unfavourable) energies of interaction at these two subsites, and may be related to the greater conformational flexibility of arabinofuranosyl residues than glycopyranosyl residues. The complete subsite maps are also rationalized with regard to the observed action patterns of these enzymes on linear (1-->5)-alpha-L arabinan. PMID- 9352636 TI - Study of the action of human salivary alpha-amylase on 2-chloro-4-nitrophenyl alpha-maltotrioside in the presence of potassium thiocyanate. AB - The degradation mechanism of a synthetic substrate, 2-chloro-4-nitrophenyl alpha maltotrioside (CNP-G3), by human salivary alpha-amylase (HSA) was investigated by kinetic and product analyses. It was observed that the enzyme attacked the various CNP-maltooligosaccharides (CNP-G3 to CNP-G6) releasing free CNP. Addition of 500 mM potassium thiocyanate (KSCN) was also found to greatly increase the rates of CNP-release. It was the fastest with CNP-G3, and, in the presence of KSCN, was almost comparable to that of degradation of maltopentaose (G5). On the other hand, addition of KSCN decreased the rate of cleavage between glucan-glucan bonds in maltopentaose. Product analysis showed that KSCN addition altered the cleavage distribution which occurred 100% at the bond between CNP and G3, and that product distribution of free CNP was largely dependent on substrate concentration. Formation of CNP-G6, a larger product than the original substrate CNP-G3, was found to be present in the digest at high concentrations of substrate and in the presence of KSCN. Based on these results, a degradation pathway for CNP-G3 involving transglycosylation besides direct hydrolysis is proposed. The increase of the CNP-release by the addition of KSCN would result from a corresponding increase in the interaction between the CNP moiety and the corresponding subsite near the catalytic site, as well as the enhancement of the catalytic efficiency. PMID- 9352637 TI - A new family of oligosaccharides from the xyloglucan of Hymenaea courbaril L. (Leguminosae) cotyledons. AB - The xyloglucan from cotyledons of Hymenaea courbaril was hydrolysed with endo (1,4)-beta-D-glucanase (cellulase) and analysed by TLC and HPAEC. The limit digest was different from those obtained from xyloglucans of Tamarindus indica and Copaifera langsdorffii. On treatment with nasturtium beta-galactosidase, two main oligosaccharides were detected by TLC and HPAEC. Using a process of enzymatic sequencing involving alternate treatments with a pure xyloglucan oligosaccharide-specific alpha-xylosidase, and a pure beta-glucosidase, both from nasturtium, their structures were deduced to be XXXG and a new oligosaccharide XXXXG. These structures were confirmed by 1H NMR. The relative proportions of XXXG and XXXXG indicate that approximately half of the subunits in Hymenaea xyloglucan are based on the new oligosaccharides. In the native polymer the XXXXG subunits are likely to carry galactosyl substituents in varying proportions, since cellulase hydrolysates contained many bands which were converted to XXXXG on hydrolysis with nasturtium beta-galactosidase. Although no comparative studies on the physico-chemical properties of Hymenaea courbaril xyloglucan have yet been performed, our results indicate that this polymer is less interactive with iodine when compared with T. indica and C. langsdorffii xyloglucans, suggesting that changes in conformation may occur due to the presence of XXXXG. PMID- 9352638 TI - Structural studies of the O-specific side-chain of lipopolysaccharide from Burkholderia gladioli pv. gladioli strain NCPPB 1891. AB - A polymeric fraction (the O-antigenic side-chain) has been isolated from the lipopolysaccharide of Burkholderia gladioli pv. gladioli strain NCPPB 1891 after mild acid hydrolysis. The components of the polymer and their molar proportions were L-Rha (1), D-Gal (1), D-Man (1), and O-acetyl (1). By means of chemical degradations and NMR studies, the repeating unit of the polymer was shown to be a linear trisaccharide of the structure shown. [formula: see text] PMID- 9352639 TI - A scanning force- and fluorescence light microscopy study of the structure and function of a model pulmonary surfactant. AB - The structure of an artificial pulmonary surfactant was studied by scanning force and fluorescence light microscopy (SFM, and FLM, respectively). The surfactant- a mixture of dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG) and recombinant surfactant-associated protein C (SP-C)--was prepared at the air-water interface of a Langmuir film balance and imaged by FLM under various states of compression. In order to visualize their topography by SFM, the films were transferred onto a solid mica support by the Langmuir-Blodgett (LB) technique. We found that a region of high film compressibility of the spread monolayer close to its equilibrium surface pressure (pi = 50 mN/m) was due to the exclusion of layered protrusions with each layer 5.5 to 6.5 nm thick. They remained associated with the monolayer and readily reinserted upon expansion of the film. Comparison with the FLM showed that the protrusions contained the protein in high concentration. The more the film was compressed, the larger was the number of layers on top of each other. The protrusions arose from regions of the monolayer with a distinct microstructure that may have been responsible for their formation. The molecular architecture of the microstructure remains to be elucidated, although some of it can be inferred from spectroscopic data in combination with the SFM topographical images. We illustrate our current understanding of the film structure with a molecular model. PMID- 9352640 TI - The phase behavior of lipid monolayers containing pulmonary surfactant protein C studied by fluorescence light microscopy. AB - Three compounds of the pulmonary surfactant--dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), and the surfactant associated protein C (SP-C)--were spread at the air-water interface of a Langmuir trough as a model system to mimic the properties of natural surfactant. Fluorescence microscopical images of the film formed at the interface were obtained during compression using a fluorescence dye bound covalently either to phosphatidylcholine or to SP-C. The images were quantified using statistical methods in respect to relative areas and relative fluorescence intensities of the domains found. In the early stage of compression, film pressure rose slightly and was accompanied by a phase separation which could be recognized in the images by the formation of bright and dark domains. On further compression, after a steep increase of film pressure, a plateau region of constant film pressure started abruptly. During compression in the plateau region, fluorescence intensity of the bright domain formed in the early stage of compression increased. The increasing fluorescence intensity, the non-Gaussian intensity distribution of the bright domain, and the small mean molecular area of the film in the plateau region gave rise to the assumption that multilayer structures were formed in the late stage of compression. The formation of the multilayer structures was fully reversible in repeated compression-expansion cycles including the plateau region of the phase diagram. The ability of lipid/SP-C mixtures to form reversible multilayer structures during compression may be relevant to stability in lungs during expiration and inhalation. PMID- 9352641 TI - A minichromosome carrying a pigmentation gene and brook trout DNA sequences in transgenic rainbow trout. AB - We describe the transmission of an introduced minichromosome of brook trout (Salvelinus fontinalis) origin, carrying a pigmentation gene, through three generations in rainbow trout (Oncorhynchus mykiss). The minichromosome was originally introduced into gynogenetic albino rainbow trout using gamma irradiated brook trout sperm. In the third generation, the presence of the minichromosome was correlated with pigmentation. A brook trout specific interspersed repeat DNA sequence, Fok I, was also correlated with pigmentation in these individuals. This system, the first clearly documented example of induced chromosome mediated gene transfer at the organismal level, could have applications in studies of gene mapping, development, gene regulation, and chromosome function. PMID- 9352642 TI - RAPD analysis of genetic variation in the Australian fan flower, Scaevola. AB - The use of randomly amplified polymorphic DNA (RAPD) to study genetic variability in Scaevola (family Goodeniaceae), a native Australian species used in ornamental horticulture, is demonstrated. Plants of the genus Scaevola are commonly known as "fan flowers," due to the fan-like shape of the flowers. Nineteen accessions of Scaevola (12 cultivated and 7 wild) were studied using 20 random decamer arbitrary primers. Eight primers gave a distinct reproducible amplification profile of 90 scorable polymorphic fragments, enabling the differentiation of the Scaevola accessions. RAPD amplification of genomic DNA revealed a high genetic variability among the different species of Scaevola studied. Molecular markers were used to calculate the similarity coefficients, which were then used for determining genetic distances between each of the accessions. Based on genetic distances, a dendrogram was constructed. Though the dendrogram is in general agreement with the taxonomy, it also highlights discrepancies in the classification. The RAPD data showed that Scaevola aemula (series Pogogynae) is closer to Scaevola glandulifera of series Globuliferae than to the rest of members of series Pogogynae. In addition, the RAPD banding pattern of white flower S. aemula, one of the commercial cultivars, was identical to that of Scaevola albida, indicating their genetic similarity. Our study showed that there is a large genetic distance between commercial cultivars of Scaevola (Purple Fanfare, Pink Perfection, and Mauve Cluster), indicating considerable genetic variation among them. The use of RAPDs in intra- and inter-specific breeding of Scaevola is also explored. PMID- 9352643 TI - Characterization and genomic organization of Ty1-copia group retrotransposons in rye (Secale cereale). AB - The genomic organisation of the Ty1-copia retrotransposons in rye (Secale cereale) has been studied. We have used the polymerase chain reaction (PCR) to amplify sequences from a conserved domain of the reverse transcriptase gene of the Ty1-copia retrotransposons in this species. Sequence analysis of 26 of these PCR products shows them to be a highly heterogeneous population, a feature that is common in plants. Slot blot analysis shows that there are about 100,000 individual Ty1-copia retrotransposons in rye. In situ hybridization of a heterogeneous probe, representing the whole population of rye Ty1-copia retrotransposon sequences, to chromosome spreads of triticale (xTriticosecale), a rye-wheat hybrid, shows that these sequences are present throughout all the rye chromosomes but absent from the centromeric regions and, in particular, from the terminal heterochromatin. Southern analysis of oat, barley, wheat, and rye, using as a probe R9, one of the rye sequences that is closely similar to the BARE-1 element of barley, shows that close relatives of this retrotransposon subgroup are present in all these species in high copy number. Northern analysis on RNAs from seedlings shows that the BARE-1 subgroup is transcribed in all these cereal plants but in variable amounts: high in barley, moderate in wheat and rye, and extremely low in oat. PMID- 9352644 TI - Physical localisation of repetitive DNA sequences in Alstroemeria: karyotyping of two species with species-specific and ribosomal DNA. AB - Fluorescence in situ hybridization (FISH) was used to localise two species specific repetitive DNA sequences, A001-I and D32-13, and two highly conserved 25S and 5S rDNA sequences on the metaphase chromosomes of two species of Alstroemeria. The Chilean species, Alstroemeria aurea (2n = 16), has abundant constitutive heterochromatin, whereas the Brazilian species, Alstroemeria inodora, has hardly any heterochromatin. The A. aurea specific A001-I probe hybridized specifically to the C-band regions on all chromosomes. The FISH patterns on A. inodora chromosomes using species-specific probe D32-13 resembled the C-banding pattern and the A001-I pattern on A. aurea chromosomes. There were notable differences in number and distribution of rDNA sites between the two species. The 25S rDNA probe revealed 16 sites in A. aurea that closely colocalised with A001-I sites and 12 in A. inodora that were predominantly detected in the centromeric regions. FISH karyotypes of the two Alstroemeria species were constructed accordingly, enabling full identification of all individual chromosomes. These FISH karyotypes will be useful for monitoring the chromosomes of both Alstroemeria species in hybrids and backcross derivatives. PMID- 9352645 TI - Map-based cloning of a gene sequence encoding a nucleotide-binding domain and a leucine-rich region at the Cre3 nematode resistance locus of wheat. AB - The Cre3 gene confers a high level of resistance to the root endoparasitic nematode Heterodera avenae in wheat. A DNA marker cosegregating with H. avenae resistance was used as an entry point for map-based cloning of a disease resistance gene family at the Cre3 locus. Two related gene sequences have been analysed at the Cre3 locus. One, identified as a cDNA clone, encodes a polypeptide with a nucleotide binding site (NBS) and a leucine-rich region; this member of the disease resistance gene family is expressed in roots. A second Cre3 gene sequence, cloned as genomic DNA, appears to be a pseudogene, with a frame shift caused by a deletion event. These two genes, related to members of the cytoplasmic NBS-leucine rich repeat class of plant disease resistance genes were physically mapped to the distal 0.06 fragment of the long arm of wheat chromosome 2D and cosegregated with nematode resistance. PMID- 9352646 TI - Ribosomal RNA genes specific to the B chromosomes in Brachycome dichromosomatica are not transcribed in leaf tissue. AB - Ribosomal RNA genes are present near the end of the short arm and, to a lesser extent, near the centromere of the B chromosomes of some populations of Brachycome dichromosomatica. The internal transcribed spacer (ITS2) was amplified by PCR from total leaf DNA using primers within the conserved regions encoding the 5.8S and 25S stable rRNA species. Comparison of PCR amplified ITS2 sequences from several individual plants without B chromosomes with corresponding sequences derived from microdissected B chromosomes revealed two consistent differences between the rDNA of A and B chromosomes. One of these differences produced an SfcI restriction site that was present only in the ITS2 of the B-chromosome rDNA. Amplification by PCR of ITS2 from total genomic DNA from plants with and without B chromosomes showed an additive relationship between the amount of PCR product containing the SfcI site and the number of B chromosomes present. Quantitative analysis indicated that the proportion of total nuclear rDNA present on a single B chromosome varied between 2 and 4% in different A chromosome backgrounds. Similar experiments, with appropriate positive and negative controls, using reverse transcriptase PCR of the equivalent region within the 40S precursor rRNA, suggested that the B-chromosome rDNA was not transcribed. Similarly, PCR of reverse transcribed total RNA from plants containing B chromosomes using primers specific for the B chromosome ITS2 was unable to detect a transcript from the B chromosome. PMID- 9352647 TI - Laser isolation of plant sex chromosomes: studies on the DNA composition of the X and Y sex chromosomes of Silene latifolia. AB - X and Y sex chromosomes from the dioecious plant Silene latifolia (white campion) were isolated from mitotic metaphase chromosome preparations on polyester membranes. Autosomes were ablated using an argon ion laser microbeam and isolated sex chromosomes were then recovered on excised fragments of polyester membrane. Sex chromosome associated DNA sequences were amplified using the degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) and pools of DOP-PCR products were used to investigate the genomic organization of the S. latifolia sex chromosomes. The chromosomal locations of cloned sex chromosome repeat sequences were analysed by fluorescence in situ hybridization and data complementary to laser ablation studies were obtained by genomic in situ hybridization. In combination, these studies demonstrate that the X and Y sex chromosomes of S. latifolia are of very similar DNA composition and also that they share a significant repetitive DNA content with the autosomes. The evolution of sex chromosomes in Silene is discussed and compared with that in another dioecious species, Rumex acetosa. PMID- 9352648 TI - On the origins of the tetraploid Bromus species (section Bromus, Poaceae): insights from internal transcribed spacer sequences of nuclear ribosomal DNA. AB - The internal transcribed spacer (ITS) region of nuclear ribosomal DNA from 22 diploid and tetraploid annual Bromus species of section Bromus (Poaceae) and three species belonging to other Bromus sections, Bromus catharticus (section Ceratochloa), Bromus anomalus (section Pnigma), and Bromus sterilis (section Genea), were investigated by PCR amplification and direct sequencing. The length of the ITS-1 region varied from 215 to 218 bp, and that of the ITS-2 region from 215 to 216 bp, in the species analyzed. ITS-1 was more variable and provided more informative sites (49) than ITS-2 (32). No variation was encountered within species. In pairwise comparison among species of section Bromus, sequence divergence ranged from 0.0 to 8.0% for the combined ITS-1 and ITS-2 regions. Parsimony analysis using Avena longiglumis and Hordeum vulgare as outgroups resulted in well-resolved phylogenetic trees and showed that section Bromus is monophyletic according to the species analyzed outside of the section. The analysis clarified the phylogenetic relationships among monogenomic (diploid) species. Introduction of the allotetraploid species did not change the general topology of the trees obtained using only the diploid species. Although some tetraploid-diploid species relationships will have to be clarified with faster evolving markers, the ITS sequences are shown to be useful for assessing evolutionary relationships among closely related Bromus species, as well as for clarifying taxonomic problems in previously controversial cases (e.g., Bromus alopecuros and Bromus caroli-henrici). New hypotheses are proposed concerning the origin of several allotetraploid species. For example, it is shown that the tetraploid Bromus hordeaceus diverged earlier than all other species of section Bromus, excluding the diploid B. caroli-henrici, which is found to be basal in this group. The tetraploid Bromus arenarius, which was considered a hybrid between sections Bromus and Genea, and the tetraploid Bromus adoensis are sister taxa within section Bromus; they belong in a weakly differentiated clade with the diploids Bromus brachystachys, Bromus japonicus, Bromus squarrosus, Bromus arvensis, and Bromus intermedius. PMID- 9352649 TI - Identical megabase transgenes on mouse chromosomes 3 and 4 do not promote ectopic pairing or synapsis at meiosis. AB - To investigate ectopic interactions at the chromatin level, we examined the meiotic organization of 1-2 mb phage lambda transgenes on mouse chromosomes 3 and 4 by fluorescence in situ hybridization in combination with immunocytology of meiotic chromosomes. At early meiotic prophase, the transgenes are sufficiently dispersed in the nuclear volume to permit potential DNA-DNA interactions, but no synaptonemal complexes form between the sites of transgenes residing on different chromosomes. At later stages, when the chromatin is more condensed, the transgenes on different chromosomes are not preferentially associated as they are when they are on the same chromosome. At diplotene and metaphase I, no formations were observed that could be interpreted as reciprocal crossovers or chiasmata between the transgenes located on chromosomes 3 and 4. It appears that in normal fertile mice, a I- to 2-mb homology is insufficient to initiate synapsis between nonhomologs, and it is concluded that homology is assessed within the broader context of the chromosome to initiate synapsis at meiotic prophase. PMID- 9352650 TI - A family of centromeric satellite DNAs from the European brown frog Rana graeca italica. AB - Digestion of Rana graeca italica DNA with Asp 718I produces highly repetitive fragments of 281 and 385 bp that were cloned and sequenced. The shorter fragment corresponds to the unit repeat (RgiS1b) of a satellite DNA. The longer fragment was found to be part of a 494-bp repeat of another satellite DNA (RgiS1a) that was cloned intact as an EcoRV fragment. RgiS1b is 97% homologous to RgiS1a, from which it seems to be derived by a single deletion. Among all species tested, only the related brown frog Rana dalmatina contained homologous repetitive DNA. The overall number of RgiS1a and RgiS1b repeats per R. graeca italica haploid genome was estimated to be 2.7 x 10(5). RgiS1a and RgiS1b repeats are organized in separate arrays, but repetitive units formed by various combinations of the two repeats were also observed on Southern blots. The amount of these extra repeats varies greatly among animals from the same population, representing a rare case of individual variability in the satellite DNA organization. FISH with probes specific for both satellites, or for RgiS1a only, labeled the centromeric and pericentromeric heterochromatin of all chromosomes. This indicated that RgiS1a and RgiS1b are interspersed within the same heterochromatic regions of the chromosomes. PMID- 9352651 TI - Effects of long-term hypergravity on muscle, heart and lung structure of mice. AB - Quantitative changes in lung, heart and muscle structure were assessed in mice exposed for 14 weeks to a gravitational field of 3 G since the age of 4 weeks; matched controls were kept at normal gravity (1 G). The body mass of 3-G-exposed mice was significantly reduced by 9%, while total skeletal muscle mass remained the same fraction of body mass. The mass of the soleus muscle was found to be significantly larger in 3-G-exposed mice both in absolute (+27%) and body mass specific terms (+42%). Capillary density was significantly reduced by 22% because of a relatively larger increase of fiber cross-sectional area (+47%) than of capillary to fiber ratio (+16%). Other morphometric variables remained unchanged with hypergravity. Heart mass and mitochondrial volume were both larger in 3-G exposed mice (+15% and +27%, respectively). This difference reached statistical significance when normalized to body mass. The only significant difference in lung structure detectable by morphometric methods were a smaller volume (-9%), that paralleled lower body mass, and thinner alveolar septa (-12%). From these results it is concluded that the lung's support structures in mice are sufficiently strong to withstand the stress of long-term hypergravity; however, 3 G exposure leads to a selective hypertrophy of soleus muscle fibers while absolute capillary length in this muscle remains unaltered. PMID- 9352652 TI - Binding of excess cadmium(II) to Cd7-metallothionein from recombinant mouse Zn7 metallothionein 1. UV-VIS absorption and circular dichroism studies and theoretical location approach by surface accessibility analysis. AB - A mouse metallotbionein (MT) 1 expression system has been constructed that renders recombinant MT as a high purity Zn-coordinated protein. Spectral changes in absorption and circular dichroism following the addition of up to 7 mol equivalents of Cd2+ to recombinant Zn7-MT showed that it behaves like the native protein. Exposure of Cd7-MT to Cd2+ resulted in further binding of these ions to the protein, although saturation was not achieved on the addition of up to 22 mol equivalents of Cd2+ to Zn7-MT. Spectral data are compatible with a model in which the first four additional Cd2+ ions are bound to Cd7-MT via sulfur atoms, and indicate that no further thiol groups are involved in the binding of the excess Cd(II) over 11. Cd2+ ions bound in excess to Cd7-MT appear to have lower binding constants as exposure of Cdn-MT (n > 7) species to Cbelex-100 retrieved Cd7-MT. Based on the X-ray data, the accessible surface areas of sulfur atoms in Cd5,Zn2 MT 2 were calculated. This led us to propose that the coordination of the first three additional Cd(II) ions to Cd7-MT proceeds by means of S-Met1-O-Met1, S-Cys7 S-Cys13 and S-Cys5-S-Cys26 pairs. Finally, comparison of the behavior of the entire MT with that of the recombinant alpha MT and beta MT subunits indicates that mutual influences may not be negligible. PMID- 9352653 TI - The Soret circular dichroism spectrum as a probe for the heme Fe(III)-Met(80) axial bond in horse cytochrome c. AB - A spectroscopic signal sensitive to the strength of the heme iron(III)-Met(80) bond in cytochrome c represents a useful tool that will provide valuable information on the heme pocket region and redox properties of the protein. At present, the 695-nm absorption band is perhaps the simplest diagnostic signal for the axial bond; this band disappears when the Fe(III)-Met(80) bond is disrupted. From the analysis of the Soret region circular dichroism spectrum of cytochrome c under conditions that gradually induce disruption of the Fe(III)-Met(80) bond, we present evidence that the 416-nm spectral dichroic band provides independent information addressing the strength of the axial bond between Fe(III) and Met(80) in cytochrome c. Further, this study demonstrates extension of the diagnostic application to very dilute protein samples based on the useful sample concentration of 5-10 microM vs 200-300 microM required for 695-nm absorbance measurements. PMID- 9352655 TI - Complexation of copper(I) by thioamino acids. Implications for copper speciation in blood plasma. AB - There is mounting evidence that Cu(I) is the most important oxidation state of copper in many physiological systems. Research into Cu(I)-thioamino acid complex formation serves not only to improve the chelation therapy for treating copper intoxication but may also provide a better understanding of many facets of normal copper metabolism. Formation constants for the ternary mixed ligand complexes of Cu(I) with cysteine (Cys), glutathione (GSH) and penicillamine (Pen) are reported here for the first time. Potentiometric titrations, using techniques specially developed for the stabilization of aqueous Cu(I), were performed at 25 degrees C in 1.00 M (Na)Cl. It was found that precipitation severely limits the experimentally accessible pH range and, consequently, the computer analysis of the binary metal-ligand systems; however, it is also found that this is less of a problem when two different ligands are present. This latter fact permitted better models of the binary systems to be developed. The formation constants of Cu(I) thioamino acids determined in this work were used in an improved computer simulation of copper speciation in blood plasma which, for the first time, incorporates redox equilibria. PMID- 9352654 TI - Bis-tropolonato derivatives of cobalt(III) complexes of bidentate aliphatic nitrogen mustards as potential hypoxia-selective cytotoxins. AB - A series of cobalt(III) complexes, [Co(trop)2(L)]+, where trop is the tropolonate anion and L is a bidentate amine or nitrogen mustard, have been prepared as potential hypoxia-selective cytotoxins (L = BEE, N,N'-diethylethylenediamine; DEE, N,N-diethylethylenediamine; BCE, N,N'-bis(2-chloroethyl)ethylenediamine; DCE, N,N-bis(2-chloroethyl)ethylenediamine). The 1H NMR and 13C{1H} NMR spectra of the complexes were assigned on the basis of chemical shift considerations, 2D NMR studies (including 13C-1H and 1H-1H COSY correlation experiments), and comparison to the related, known acetylacetonato (acac) complexes [Co(acac)2(L)]+. An x-ray crystal structure determination of the analogue [Co(trop)2(BEE)]ClO4 showed it to be the delta SS/lambda RR enantiomeric pair of diastereomers. The tropolonato complexes have significantly higher reduction potentials than the corresponding acac complexes, suggesting more facile cellular reduction. In agreement with this, the mustard complexes have IC50 values in cells little different to those of the free mustards even under aerobic conditions, and do not show hypoxic selectivity in a clonogenic assay under conditions where the corresponding 3-methylacac complex of DCE showed significant selectivity. PMID- 9352656 TI - Comparison of exopolyphosphatases of different yeast cell compartments. AB - Purified cell-envelope polyphosphatase as well as polyphoshatase activities of cytosol and isolated vacuoles, of nuclei and mitochondria of the yeast Saccharomyces cerevisiae were compared. The polyphosphatases of cell envelope and cytosol are similar, the polyphosphatases of nuclei, vacuoles and mitochondria differ in their kinetic properties, substrate specificity, requirements in divalent cations and in some effector actions both from these and from each other. PMID- 9352657 TI - Penicillin-binding protein sensitive to cephalexin in sporulation of Bacillus cereus. AB - Cephalexin, cefaclor, cefadroxil, and cefotaxime strongly inhibited sporulation of Bacillus cereus ts-4 at 1 microgram/ml. Cephalexin was most inhibitory on the sporulation of B. cereus when the antibiotic was added at 3 h after induction of sporulation by nutrient downshift technique. Examination of 4',6-diamidino-2 phenylindole-stained cells by fluorescence-phase contrast microscopy showed that cephalexin inhibited the formation of asymmetric septum. By using [3H]penicillin, eight penicillin-binding proteins (PBPs) were detected from the cells of B. cereus ts-4. Among them, four PBPs were also detected in sporulating cells. Affinity of cephalexin to PBPs were measured indirectly by competition for subsequent binding of radioactive penicillin G. Cephalexin strongly bound to PBP 4 with molecular weight of 72,000 in sporulating cells. PMID- 9352658 TI - Rapid physiological characterization of microorganisms by biosensor technique. AB - Eleven microorganisms, Arxula adeninivorans LS3, Candida boidinii DSM 70034, Candida lactis-condensi DSM 70635, Pichia jadinii DSM 2361, Pichia minuta DSM 7018, Kluyveromyces lactis DSM 4394, Pseudomonas putida DSM 50026, Alcaligenes sp. DSM 30002, Arthrobacter nicotianae DSM 20123 as well as Issatchenkia orientalis DSM 70077 and Rhodococcus erythropolis DSM 311 were characterized by the sensor technique by injection of 30 different substrates and substrate mixtures. The obtained data which are based on the determination of respiratory rate of microorganisms are similar to physiological characteristics obtained with conventional methods. In comparison to these conventional methods the sensor technique works much more rapid and permits quantification of the data. Therefore, the described technique provides an alternative method for the characterization of microorganisms. PMID- 9352659 TI - Antimicrobial action of propolis and some of its components: the effects on growth, membrane potential and motility of bacteria. AB - The effect of the natural bee product propolis on the physiology of microorganisms was investigated using B. subtilis, E. coli and R. sphaeroides. An ethanolic extract of propolis had a bactericidal effect caused by the presence of very active, but labile, ingredients. The exact bactericidal effect of propolis was species dependent: it was effective against gram-positive and some gram negative bacteria. Propolis and some of its cinnamic and flavonoid components were found to uncouple the energy transducing cytoplasmic membrane and to inhibit bacterial motility. These effects on the bioenergetic status of the membrane may contribute to the antimicrobial action of propolis and its observed synergism with selected antibiotics. PMID- 9352660 TI - Leukotoxic factors produced by staphylococci of ovine origin. AB - Leukotoxins produced by staphylococci, especially Staphylococcus aureus, have long been considered important virulence determinants. The present assay examined 26 strains of staphylococci isolated from sheep for their ability to produce factors leukotoxic to polymorphonuclear leukocytes deriving from ovine mammary glands. Twenty one strains were coagulase-negative staphylococci and five coagulase-positive. Of the coagulase-negative staphylococci 17 were isolated from subclinical mastitis and four from the teat skin. The coagulase-positive strains were isolated from clinical mastitis and caused death to 91-100% of the cells. The coagulase-negative strains isolated from subclinical mastitis caused death to less than 50% of cells, while those from the skin did not affect any cells. The concentration of active leukotoxins is an important staphylococcal virulence factor. Low production of leukotoxins or no production by coagulase-negative staphylococci may determine their ability to survive the mammary gland defenses or cause clinical mastitis. PMID- 9352661 TI - Differential amplification efficiency of pMB1 and p15A (ColE1-type) replicons in Escherichia coli stringent and relaxed strains starved for particular amino acids. AB - It was demonstrated previously that ColE1-type plasmids, the most commonly used vectors in molecular cloning, can be amplified in amino acid-starved relA mutants of Escherichia coli. Subsequent studies demonstrated that replication of at least some plasmids during amino acid starvation depends not only on the host relA allele but also on temperature and on the nature of deprived amino acid. Therefore, we investigated efficiency of amplification of two types of ColE1 plasmids (pMB1- and p15A-derived replicons) in E. coli relA+ and relA- hosts starved for different amino acids at 30 degrees C, 37 degrees C and 43 degrees C. We found differential amplification efficiency of plasmids pBR328 (pMB1-derived replicon) and pACYC184 (p15A-derived replicon) in the relA mutant during starvation for particular amino acids. Although amplification of pBR328 was negligible in the relA+ host, significant increase in pACYC184 content was observed in this strain starved for some (but not all) amino acids. The amplification efficiency of pBR328 and pACYC184 was found to be dependent on temperature. These results indicate that for maximal amplification of particular plasmid appropriate amino acid starvation and optimal temperature should be chosen. Our findings are in agreement with recently proposed model of the regulation of ColE1-type plasmid replication in amino acid-starved E. coli cells. PMID- 9352662 TI - Organisation of ribosomal DNA in the ascomycete Leptosphaeria maculans. AB - In the ascomycete Leptosphaeria maculans tandem repeats of ribosomal DNA (rDNA) are restricted to one or two particular chromosomes of the 15 chromosomes of 19 field isolates examined. Ribosomal DNA can account for size differences of 35% between homologous chromosomes in a particular tetrad. During crossing, no detectable recombination between blocks of tandem repeats, nor changes in their size occur. The organisation of rDNA in L. maculans differs from many other haploid fungi. Firstly, sequence heterogeneity occurs within tandem repeats of rDNA; regularly spaced Sal 1 sites (0.25 Mb apart) are present within a 1.4 Mb block of tandem repeats. Secondly, individual isolates have different-sized rDNA repeats; this variation occurs in the non-transcribed intergenic spacer region. Thirdly, there is a wide range in the copy number of the rDNA repeat (from 56 to 225) amongst only four field isolates examined. PMID- 9352663 TI - Involvement of IMP dehydrogenase activity in induction of sporulation of Bacillus cereus. AB - IMP dehydrogenase activity of B. cereus increased parallel to cell growth in YE EMM, where B. cereus did not sporulate. When B. cereus was cultured in a modified G medium, a sporulation medium, the activity reached the highest level at 6 hr and decreased thereafter. After induction of sporulation by nutritional shift down in 1/100 G medium, the enzyme activity decreased to about 5% compared with exponentially growing cells at 1 hr of resuspension. The sporulation rate of B. cereus was over 90% in the modified G medium and 1/100 G medium. Sporulation was strongly inhibited by mycophenolic acid at 1 mM, when the drug was added at 0 and 1 hr of resuspension in 1/100 G medium. Intracellular GTP concentration of B. cereus decreased to the lowest level about 1 hr of resuspension. Although GTP increased to about 50% of the exponentially growing cells at 2 hr of resuspension in control cells, the concentration did not increase in the presence of 1 mM mycophenolic acid. PMID- 9352664 TI - Molecular identification and relatedness of potato witches'-broom phytoplasma isolates from four potato cultivars. AB - Four isolates of potato witches'-broom phytoplasma, designated as PW1, PW2, PW3 and PW4, were established on four potato cultivars. The identity of each isolate was confirmed by PCR using two universal primer pairs and one specific primer set derived from phytoplasma of 16S rDNA sequences. The four isolate samples formed similar RFLP patterns after digestion of 1.2 kb PCR products with restriction endonucleases AluI, HhaI, RsaI and Sau3A. The direct DNA sequencing with the specific primer pair showed that there are no differences in the base sequences among PW1, PW2, and PW3 phytoplasma isolates and that PW4 is closely related to them. Thus, the four isolates were identified as members of the clover proliferation group. PMID- 9352665 TI - Aetiology of yam (Dioscorea rotundata) tuber rots held in traditional stores in Nigeria: importance of Fusarium spp. and yam beetle. AB - The extent and causes of yam (Dioscorea rotundata) tuber rots were investigated in Igalaland, Nigeria. Rots were found to be associated with prior physical damage in almost all cases. Numerous fungal species were isolated from the infected lesions of which Fusarium spp. predominated, not Botrydiplodia theobromae as previously reported in other studies. None of the fungi was able to infect undamaged yams in laboratory experiments. The damage was caused by both biotic and abiotic factors of which the yam beetle (Heteroligus meles) was the largest single cause. There were significant differences in the frequency with which different varieties of yam were attacked by H. meles and found to be infected with Fusarium spp. In an independent survey of farmers' experience of yam rots, varieties that scored well correlated with apparent resistance to the beetle and Fusarium spp. These studies highlight the importance of yam beetle infestation and the need to have independent methods for varietal authentication. PMID- 9352667 TI - Laccase-catalyzed formation of cinnabarinic acid is responsible for antibacterial activity of Pycnoporus cinnabarinus. AB - Concentrated culture fluid of the wood-rotting basidiomycete Pycnoporus cinnabarinus showed biological activity against a variety of bacterial strains. The maximal inhibitory effect was obtained for Gram-positive bacteria of the genus Streptococcus. In general, inhibition was higher for Gram-positive than Gram-negative bacteria. P. cinnabarinus produces the phenoxazinone derivative, cinnabarinic acid. This red pigment accumulates in sporocarps as well as in liquid cultures. As shown previously, laccase secreted by the fungus oxidizes the precursor 3-hydroxyanthranilic acid to cinnabarinic acid. The present study demonstrates that this reaction is necessary for the production of antibacterial compounds by the fungus. The biological activity of concentrated P. cinnabarinus culture fluid was nearly identical with that of cinnabarinic acid, synthesized by purified laccase in vitro. PMID- 9352666 TI - Electrophoretic analysis of hydrolytic enzymes of Escherichia coli cells starved in seawater and drinking water: comparison of gelatinolytic, caseinolytic, phosphohydrolytic and hyaluronolytic activities. AB - Starvation of four Escherichia coli clinical strains in seawater and drinking water for nine days revealed that various changes of hydrolytic enzymes were induced. Several gelatinolytic and caseinolytic activities differing in mol mass were detected both in seawater and drinking water starved cells by substrate gel electrophoresis. The major activities of gelatinase migrated with mol masses of approximately 170 kDa and approximately 45 kDa. On the contrary, hyaluronolytic activities were detected only in cells cultured in Mueller Hinton broth with average mol masses of 36 kDa and 45 kDa. Acid and alkaline phosphohydrolytic activities were detected by native electrophoresis. Both activities were decreased in number of bands in E. coli cells starved either in seawater or drinking water. PMID- 9352668 TI - Aluminium, iron and manganese in near-surface waters of a tropical rainforest ecosystem. AB - A hydrochemical investigation was undertaken in a tropical rainforest catchment to determine the aluminium, iron, and manganese concentrations in near-surface waters of a non-impacted, circum-neutral environment, with an emphasis on hydrologic controls of Al, Fe, and Mn stormflow variations. The concentrations of all three solutes increase with discharge, with antecedent moisture conditions influencing the amplitude. This increase is accompanied by a minor pH depression from about 7.3 to 6.8, and, in the case of one event, by an increase in DOC. For all three solutes, the ranges in stormflow and overland flow concentrations are nearly identical. Al, Fe, and Mn concentrations in soil and ground water are considerably lower, with the exception of Mn in shallow soil water which is similar to overland flow. We conclude that episodic increases in streamflow metal concentrations in this tropical rainforest environment are not so much the result of a pronounced pH depression, but of an overland flow-mediated input from near surface sources such as leaf litter and topsoil. PMID- 9352669 TI - Nutrient dynamics in a lowland stream impacted by sewage effluent: Great Ouse, England. AB - Nutrient and ancillary chemical changes in a stretch of the Great Ouse river near Brackley in Northamptonshire, UK, were measured on a seasonal basis over one year with an initial pilot study in the spring of 1994. River bed-sediments were characterized by their physical, adsorptive and chemical properties and a batch returned to the laboratory for experiments in a fluvarium channel. These experiments involved studies of the release and uptake of soluble reactive phosphorus to or from the overlying water in oxic conditions and release when the dissolved oxygen concentration was reduced to near zero. There was no impact of the point-sources on the concentration of nitrate in the river, a slight effect on the concentration of silicon during low-flow conditions in the summer and net uptake in the spring caused by the growth of diatoms. However there was a substantial impact on phosphorus concentrations, particularly during the summer sampling when the river was in low flow. The results for the winter showed little impact of point discharges because of the high dilution of the treated effluent. The bed-sediments at this time were found to be close to equilibrium with respect to the concentration of soluble reactive phosphorus in the overlying water. Both the fluvarium channel and field measurements obtained in the autumn are consistent with a lower net uptake of phosphorus and degradation of vegetation in the river. In the spring and summer visits, the phosphorus concentrations increased immediately downstream of the main point input and then decreased in concentration at the next downstream site. This effect was particularly noticeable in the summer and was consistent with a large uptake of phosphorus to the bed-sediment and associated vegetation. The contribution of the bed-sediment was estimated using a chemical model describing the uptake kinetics by the Elovich equation and also a parabolic equation. The stability of the waters with respect to calcite and calcium phosphate minerals was assessed in detail. Seasonal changes in the sediment composition were consistent with the deposition of calcite and coprecipitation of inorganic phosphate in the lattice of calcite, either abiotically, or in association with algal biofilms in the sediment. Good correlation between the total phosphorus and calcium contents of the sediments were evident, particularly at the sites furthest from the main sewage input. Measurements of the equilibrium phosphate concentrations of the surface sediments showed that they did not respond quickly to the higher concentrations of dissolved phosphorus found in the summer. It is also evident that the use of the equilibrium phosphate concentration to predict the magnitude of the release of soluble reactive phosphorus becomes less reliable as the solution concentration approaches the equilibrium phosphate concentration. Perturbations may arise because of changes in the surface micro-layer caused by a number of processes such as particle-size fractionation, biological activity or changes in the local redox conditions. However bearing these constraints in mind, with an equilibrium phosphate concentration of the sediments generally below 5 mumol dm-3, the release of phosphorus to the overlying water is not expected until the concentration is below this value. The results also show that phosphorus is not accumulating in the surface sediment and that much of the phosphorus in the sediment is not easily desorbed. PMID- 9352670 TI - Quantifying health effects from the combined action of low-level radiation and other environmental agents: can new approaches solve the enigma? AB - Efforts to assess the quantify deleterious effects from toxicants are directed mainly towards single agents, whereas real world environmental and occupational exposures to natural and anthropogenic agents quite often entail the concomitant presence of several toxicants. These combined exposures may lead to health risks that differ from those expected from simple addition of the individual risks. For example, combined exposures to physical and chemical agents such as radon and smoking or asbestos and smoking produce over-additive effects at exposure levels typical for earlier workplaces. In tumour therapy, the modulation of radiation effects by cytotoxic drugs is widely used to enhance the therapeutic gain. Whether interactions occurring at high exposure levels are important at the low exposure levels set for the public and for modern workplaces is difficult to answer. A scientifically sound extrapolation from these high to low-dose levels should be based on dose-effect relationships of the relevant agents alone and in combination. In general this information is not available. The existing data base on combined effects is rudimentary, mainly descriptive and rarely covers exposure ranges large enough to make direct inferences to present day low-dose exposure situations. In view of the multitude of possible interactions between the large number of potentially harmful agents in the human environment, descriptive approaches will have to be supplemented by the use of mechanistic models for critical health endpoints such as cancer. To generalise and predict the outcome of combined exposures, agents will have to be grouped depending on their physical or chemical mode of action on the molecular and cellular level. Such a grouping must be guided by specific mechanistic studies designed to examine the underlying hypothesis regarding how various classes of agents interact. PMID- 9352672 TI - Bacterial alginates: biosynthesis and applications. AB - Alginate is a copolymer of beta-D-mannuronic acid and alpha-L-guluronic acid (GulA), linked together by 1-4 linkages. The polymer is a well-established industrial product obtained commercially by harvesting brown seaweeds. Some bacteria, mostly derived from the genus Pseudomonas and belonging to the RNA superfamily I, are also capable of producing copious amounts of this polymer as an exopolysaccharide. The molecular genetics, regulation and biochemistry of alginate biosynthesis have been particularly well characterized in the opportunistic human pathogen Pseudomonas aeruginosa, although the biochemistry of the polymerization process is still poorly understood. In the last 3 years major aspects of the molecular genetics of alginate biosynthesis in Azotobacter vinelandii have also been reported. In both organisms the immediate precursor of polymerization is GDP-mannuronic acid, and the sugar residues in this compound are polymerized into mannuronan. This uniform polymer is then further modified by acetylation at positions O-2 and/or O-3 and by epimerization of some of the residues, leading to a variable content of acetyl groups and GulA residues. In contrast, seaweed alginates are not acetylated. The nature of the epimerization steps are more complex in A. vinelandii than in P. aeruginosa, while other aspects of the biochemistry and genetics of alginate biosynthesis appear to be similar. The GulA residue content and distribution strongly affect the physicochemical properties of alginates, and the epimerization process is therefore of great interest from an applied point of view. This article presents a survey of our current knowledge of the molecular genetics and biochemistry of bacterial alginate biosynthesis, as well as of the biotechnological potential of such polymers. PMID- 9352671 TI - Nutrient profiles in the everglades: examination along the eutrophication gradient. AB - We examined the concentration profiles of nutrients in the surface water, soil and pore water along the eutrophication gradient of the Water Conservation Area 2A (WCA-2A) in the northern Everglades. Phosphorus levels in the surface waters contributed by the agricultural runoff showed an exponential decrease downstream of the inflow structures attaining background values of 7-12, 7-9 and 5-6 micrograms l-1 of TP, TDP and PO4-P, respectively, at distances of 8-10 km. The pore water PO4-P concentration in the oligotrophic areas ranged between 5 and 10 micrograms l-1. Molar ratios of dissolved inorganic N and P suggest a possible switch in nutrient limitation in the surface water from P in the oligotrophic areas to N in the eutrophic areas (DIN:DIP approximately 5). External nutrient loading has also contributed to a three- to four-fold increase in soil TP concentration and enhanced pore water PO4-P in the northern marshes. Unlike P, C and N concentration in the soils remained fairly uniform along the eutrophication gradient. 210Pb dating of soil cores suggests that the increase in soil P concentration (from < 500 to 1500 micrograms g-1) and P accumulation rate (from 0.06 to 0.46 g P m-2 per year) at the eutrophic site correlates with the installation of inflow structures in 1960-1963 through which agricultural drainage from the Hillsboro canal enters the marshes. Organic P makes up 70-90% of the total P in the soils as uptake by algae and macrophytes is the primary mechanism of P removal in these wetlands. Calcium supply from the underlying bedrock suggested from the surface and pore water chemical profiles has important consequences for P-cycling in the Everglades as Ca-bound P is the major form of inorganic P storage in the soils. PMID- 9352673 TI - Effects of ethanol concentration and stripping temperature on continuous fermentation rate. AB - The operation of a pilot plant consisting of a 14-1 fermentor, 10-cm packed column and condenser for continuous fermentation and stripping of ethanol was stable for more than 100 days. The feed consisted of a non-sterile solution of 560 g/l glucose with 100 g/l corn steep water. Fouling of the packing in the column with attached growth of yeast cells was controlled by in situ washing at intervals of 3-6 days. A computer simulation of the pilot plant was developed and used to analyze the data. The productivity of the continuous fermentor varied from 14 g ethanol to 17 g ethanol l-1 h-1. The yield was equal to the maximum theoretically possible: 0.51 g ethanol/g glucose consumed. Results are fit to linear models for the effects of ethanol concentration on specific growth rate and cell yield, and for the effect of stripping temperature on specific growth rate. PMID- 9352674 TI - Production, purification and characterization of a 50-kDa extracellular metalloprotease from Serratia marcescens. AB - The extracellular metalloprotease (SMP 6.1) produced by a soil isolate of Serratia marcescens NRRL B-23112 was purified and characterized. SMP 6.1 was purified from the culture supernatant by ammonium sulfate precipitation, acetone fractional precipitation, and preparative isoelectric focusing. SMP 6.1 has a molecular mass of approximately 50,900 Da by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). The following substrates were hydrolyzed: casein, bovine serum albumin, and hide powder. SMP 6.1 has the characteristics of a metalloprotease, a pH optimum of 10.0, and a temperature optimum of 42 degrees C. The isoelectric point of the protease is 6.1. Restoration of proteolytic activity by in-gel renaturation after SDS-PAGE indicates a single polypeptide chain. SMP 6.1 is inhibited by EDTA (9 micrograms/ml) and not inhibited by antipain dihydrochloride (120 micrograms/ml), aprotinin (4 micrograms/ml), bestatin (80 micrograms/ml), chymostatin (50 micrograms/ml), E-64 (20 micrograms/ml), leupeptin (4 micrograms/ml), Pefabloc SC (2000 micrograms/ml), pepstatin (4 micrograms/ml), phosphoramidon (660 micrograms/ml), or phenylmethylsulfonyl fluoride (400 micrograms/ml). SMP 6.1 retains full activity in the presence of SDS (1% w/v), Tween-20 (1% w/v), Triton X-100 (1% w/v), ethanol (5% v/v), and 2-mercaptoethanol (0.5% v/v). The extracellular metalloprotease SMP 6.1 differs from the serratiopeptidase (Sigma) produced by S. marcescens ATCC 27117 in the following characteristics: isoelectric point, peptide mapping and nematolytic properties. PMID- 9352676 TI - Heterologous gene expression of bovine plasmin in Lactococcus lactis. AB - Heterologous production of bovine plasmin was studied in the industrially relevant bacterium Lactococcus lactis. Two sets of lactococcal gene expression signals were coupled to the region of the plasmin gene coding for the serine protease domain. When the promoter region of the prtP gene was used, plasmin was detected mainly intracellularly in strain BPL25 by Western blot hybridization. The intracellular presence of plasmin led to physiological stress. Expression of the plasmin gene driven by the promoter and complete signal sequence of the lactococcal usp45 gene resulted in efficient plasmin secretion in strain BPL420. Cell lysis was observed in strains producing plasmin fragments including the catalytic domain, but not in control strains, which only produced a non-catalytic region of plasmin. The plasmin produced was shown to be biologically active. PMID- 9352675 TI - Parameters affecting polymerase chain reaction detection of waterborne Cryptosporidium parvum oocysts. AB - Cryptosporidium parvum is an enteric protozoan parasite of medical and veterinary importance. Dissemination of environmentally resistant oocysts in surface water plays an important role in the epidemiology of cryptospridiosis. Although the polymerase chain reaction (PCR) is a well-established technique and is widely used for detecting microorganisms, it is not routinely applied for monitoring waterborne C. parvum. In order to facilitate the application of PCR to the detection of waterborne C. parvum oocysts, a comparison of published PCR protocols was undertaken and different sample-preparation methods tested. The sensitivity of a one-step PCR method, consisting of 40 temperature cycles, was 10 purified oocysts or fewer than 100 oocysts spiked in raw lake water. The detection limit of two primer pairs, one targeting the ribosomal small subunit and another specific for a C. parvum sequence of unknown function, was approximately ten-fold lower than achieved with a primer pair targeting an oocyst shell protein gene. Three cycles of freezing/thawing were sufficient to expose oocyst DNA and resulted in higher sensitivity than proteinase K digestion, sonication or electroporation. Inhibition of PCR by surface water from different local sources was entirely associated with the soluble fraction of lake water. Membrane filtration was evaluated in bench-scale experiments as a means of removing lake water inhibitors and improving the detection limit of PCR. Using gel and membrane filtration, the molecular size of inhibitory solutes from lake water was estimated to less than 27 kDa. PMID- 9352677 TI - Improved efficiency and stability of multiple cloned gene insertions at the delta sequences of Saccharomyces cerevisiae. AB - Two delta-integration vectors were evaluated for the insertion of an inducible expression cassette (the yeast CUP1 promoter fused to the Escherichia coli lacZ structural gene, CUP1p-lacZ) and a bacterial neomycin-resistance gene (neo) into the genome of Saccharomyces cerevisiae via homologous recombination. Cells containing integrations were selected by resistance to the aminoglycoside G418. The first vector was a traditional construct containing only one delta sequence; with this vector, the transformation efficiency and the number of integrations per cell were quite low. The second carried two delta sequences flanking the desired insert, and the unneeded bacterial sequences were removed by restriction enzyme digestion immediately before transformation. When this double delta vector was employed, the integrated copy number was more than doubled relative to the single delta system and final beta-galactosidase levels exceeded those obtained with the 2 mu-based plasmid. Furthermore, the integrations appeared more stable in long-term sequential culture (both with and without induction of the lacZ gene) than those obtained via the single delta vector. PMID- 9352678 TI - Extracellular reduction of selenite by a novel marine photosynthetic bacterium. AB - A novel purple nonsulfur bacterium strain NKPB030619, which has resistance to over 5 mM selenite, was isolated from a marine environment. An initial concentration of 1.1 mM selenite, added to the medium, was decreased to under 0.05 mM within 5 days. The color of the cell suspension turned red within 2 days. The red coloration gradually decreased and black precipitates appeared during 2 weeks of cultivation. Under these conditions, two main types of deposit were formed extracellularly. These deposits were thought to contain red amorphous selenium and black vitreous selenium. The selenite reduction to elemental selenium in this bacterium was induced by the introduction of light and L-malic acid under anaerobic conditions. These results suggest that selenite reduction is coupled with photosynthesis and L-malic acid can serve as the indirect electron donor for its reduction. Phylogenetic analysis based on the 16S rDNA sequence showed that NKPB0360619 belongs to the alpha subdivision of Proteobacteria and is classified into the Rhodobacter species. The highest similarity of 86.2% was observed with R. sphaeroides. PMID- 9352679 TI - Isolation and characterization of nickel-accumulating yeasts. AB - We selected three yeast strains that efficiently remove heavy metal ions from aqueous solution. We first screened yeasts that grew in the presence of 2 mM NiCl2 among our stock of wild yeasts, and then selected those that removed Ni most efficiently from aqueous solution. These strains also removed Cu and Zn from aqueous solution and were identified as Candida species. Ni uptake was efficient at pH between 4.0 and 7.0, but less efficient at pH below 3.0. The amount of Ni taken up by the yeast cells was proportional to the initial concentration of NiCl2 below about 4 mM Ni. The cells retained the abilities to remove Ni after treatment with 10 mM EDTA or 1 M HCl for repeated usage, or after heat treatment. PMID- 9352680 TI - Removal of tetrachloroethylene in an anaerobic column bioreactor. AB - Removal of tetrachloroethylene (perchloroethylene; C2Cl4) by microbial consortia from two sites with different C2Cl4 exposure histories was examined in a bench scale anaerobic column bioreactor. It was hypothesized that optimal removal would be observed in the reactor packed with sediments having an extensive exposure history. Microbial consortia were enriched from hyporheic-zone (HZ) sediments from the Portneuf aquifer near Pocatello, Idaho, and from industrial-zone (IZ) sediments from a highly contaminated aquifer in Portland, Oregon. Lactate and acetate were the electron donors during experiments conducted over 9 and 7 months for HZ and IZ sediments, respectively. In the HZ bioreactor, the retention time ranged from 31 h to 81 h, and inlet C2Cl4 concentrations ranged from 0.1 ppm to 1.0 ppm. Dechlorination of C2Cl4 averaged 60% and reached a maximum of 78%. An increase in C:N from 27:1 to 500:1 corresponded to an 18% increase in removal efficiency. Trichloroethylene production corresponded to decreased effluent C2Cl4; further intermediates were not detected. In the IZ bioreactor, the retention time varied from 34 h to 115 h; the inlet C2Cl4 concentration was 1.0 ppm. C2Cl4 removal averaged 70% with a maximum of 98%. Trichloroethylene and cis dichloroethylene were detected in the effluent. Increases in C:N from 50:1 to 250:1 enhanced dechlorination activity. PMID- 9352681 TI - Hydrophilic surroundings requisite for the solubilization of proteins related with their hydrophobicity in the AOT reversed micellar extraction. AB - The reversed micellar extraction (AOT/isooctane system) using the phase transfer method was investigated in relation to the AOT concentration and the water solubilization for ribonuclease A, lysozyme and cytochrome c. The minimal AOT concentration required for 100% forward extraction was obtained for these proteins. At the minimal AOT concentration, the hydrophilic surroundings, i.e. the molar ratio of water to extracted protein in the organic phase, were independent of the protein concentration for each protein. The hydrophilic surroundings of these proteins were linearly related with Fisher's polarity ratio, p, as an index of the hydrophobicity of the protein. Using this linear relation, a procedure to estimate the sufficient AOT concentration for the protein extraction was proposed. In the cases of cytochrome c and lysozyme, the water concentration was larger than that in the protein-free system in spite of the same AOT condition. On the contrary, in the case of ribonuclease A, this large water uptake in the organic phase was not observed. These differences of water uptake were discussed in relation to the location of the protein in the AOT reversed micelles. PMID- 9352682 TI - An efficient three steps preparative purification of penicillin acylase from Escherichia coli cells. AB - A new and efficient safe system for the purification of the penicillin acylase from Escherichia coli G271 is presented. It was found that after a selective precipitation with ammnonium sulphate, followed by two chromatographic steps (anion exchange followed by adsorption on hydroxyapatite support), the enzyme was enriched 98 times with a 100% activity recovery. An original way has also been used to study the chromatographic separation of the protein mixture in three major categories on DEAE resin, by an analysis of the concentrations of the different species in the breakthrough curve obtained from a complete saturation of the column. PMID- 9352683 TI - Centrifugal processing of cell debris and inclusion bodies from recombinant Escherichia coli. AB - The settling characteristics of cell debris and inclusion bodies prior to, and following, fractionation in a disc-stack centrifuge were measured using Cumulative Sedimentation Analysis (CSA) and Centrifugal Disc photoSedimentation (CDS). The impact of centrifuge feedrate and repeated homogenisation on both cell debris and inclusion body collection efficiency was investigated. Increasing the normalised centrifuge feedrate (Q/sigma) from 1.32 x 10(-9) m s-1 to 3.97 x 10( 9) m s-1 leads to a 36% increase in inclusion body paste purity. Purity may also be improved by repeated homogenisation. Increasing the number of homogeniser passes results in smaller cell debris size whilst leaves inclusion body size unaltered. At a normalised centrifuge feedrate of 2.65 x 10(-9) m s-1, increasing the number of homogeniser passes from two (2) to ten (10) improved overall inclusion body paste purity by 58%. Grade-efficiency curves for both the cell debris and inclusion bodies have also been generated in this study. The data are described using an equation developed by Mannweiler (1989) with parameters of k = 0.15-0.16 and n = 2.5-2.6 for inclusion bodies, and k = 0.12-0.14 and n = 2.0-2.2 for cell debris. This is the first accurate experimentally-determined grade efficiency curve for cell debris. Previous studies have simply estimated debris grade efficiency curves using an approximate debris size distribution and grade efficiency curves determined with 'ideal particles' (e.g. spherical PVA particles). The findings of this study may be used to simulate and optimise the centrifugal fractionation of inclusion bodies from cell debris. PMID- 9352684 TI - Evaluation of affinity filters for protein isolation. AB - Affinity filters were investigated for their potential in the recovery of proteins from complex samples. The experiments covered membranes carrying high and low molecular weight affinity ligands as well as group and substance specific ones. For the ready-to-use affinity filters the specific protein binding capacity was determined and compared to that of the respective Sepharose affinity gels (Pharmacia). In the case of the pre-activated membranes the influence of the coupling chemistry on the affinity mediator concentration and the protein binding capacity were considered in the study. In the case of low molecular weight ligands (e.g. Cibacron Blue, Heparin) either type of membrane yielded stationary phases of a ligand concentration, binding capacity, resolution, and long term stability similar to that of the corresponding Pharmacia material. However, the membranes could be used at a higher flow rate than the columns, since they are less mass transfer limited and cause significantly less back pressure. The immobilization of high molecular weight ligands such as antibodies (immuno filtration) on the other hand, resulted in low ligand concentrations and worse antigen binding capacities whenever conventional immobilization procedures, e.g. epoxy group-based reactions, were used. In contradistinction, good results were obtained with tosyl- and tresyl activated membranes. Such membranes were successfully employed for the immobilization of monoclonal antibodies (mAb) and Concanavalin A. Concanavalin A and an anti gp 220/350 mAb were subsequently used to produce affinity filters for the isolation of a recombinant gp 220/350 Epstein Barr virus surface antigen from culture supernatants of a Chinese hamster ovary cell line grown in protein-free medium. PMID- 9352685 TI - Measurement of bilirubin by HPLC in bovine plasma--a comparison with the conventional diazo-method. AB - Application of a HPLC-technique according to MURACA & BLANCKAERT (1983) for accurate determination of the relative amounts of unconjugated bilirubin and its sugar mono- and diconjugates in bovine plasma is described and the results are compared with those of the conventional diazo-method. The diazo-assay yielded values for total and unconjugated bilirubin which were considerably higher than those obtained by the HPLC-technique. This is probably due to unidentified diazo positive compounds distinct from bilirubin. The direct-reacting fraction in the diazo-assay showed little or no agreement with the fraction of total ester conjugates determined by HPLC. Results indicate that the diagnostical value of bilirubin subfractioning by the diazo-method as a liver function test in cattle must be reconsidered. PMID- 9352686 TI - Alpha adrenoceptor blockade in the treatment of benign prostatic hyperplasia: past, present and future. AB - The treatment of BPH by alpha blockade is built upon a sound anatomical, physiological and pharmacological rationale. The theory is borne out in clinical practice; alpha adrenoceptor antagonists have been shown in placebo-controlled studies to improve symptoms of BPH and increase urinary flow rate. In hypertensive patients, there is a clinically significant reduction in blood pressure, with little or no effect on the blood pressure of normotensive patients with BPH. The development of selective alpha-1 adrenoceptor antagonists with a gradual onset and long duration of action has improved the tolerability and makes this class of drug a valuable alternative to surgery in many cases. Further refinements in the selectivity of alpha-1 adrenoceptor antagonists may enable even better targeted alpha blockade for BPH in the future by specific antagonism of the alpha-1 A adrenoceptor, although this hypothesis has yet to be confirmed clinically. PMID- 9352687 TI - Effects of Tamm-Horsfall protein with normal and reduced sialic acid content upon the crystallization of calcium phosphate and calcium oxalate in human urine. AB - OBJECTIVE: To examine the effects of Tamm-Horsfall protein (THP) of normal and low sialic acid content on urinary crystallization, and establish whether there are changes conducive to the formation of kidney stones. MATERIALS AND METHODS: Purified samples of THP were recovered from the urine of non-stone forming individuals. A portion of each THP sample was treated with the enzyme neuraminidase to yield the low sialic acid form of the protein. The two forms of THP were added separately to ultrafiltered urine and crystallization was then induced in the urine by evaporation at 37 degrees C. Two types of experiment were then conducted with the crystals that formed; the rate at which the resulting calcium phosphate or calcium oxalate crystals sedimented in the evaporated urine was determined and the proportion of these crystals and protein which was retained when the urine was passed through a 75 microns sieve was measured. RESULTS: Calcium phosphate and calcium oxalate crystals remained in stable colloidal suspension in ultrafiltered urine when in the presence of normal THP; these suspensions passed freely through the 75 microns sieves. When crystals formed in the presence of low sialic acid THP, the sedimentation was rapid and the crystals were readily retained with protein on the sieves. CONCLUSIONS: These results indicate that whilst normal THP inhibits urinary crystal aggregation, the properties of the low sialic acid form are consistent with the promotion of crystal aggregation and hence stone formation. PMID- 9352688 TI - Nephrogenic adenoma--a study with special reference to clinical presentation. AB - OBJECTIVE: To obtain information on the presenting symptoms, location, sex distribution, age, endoscopic appearance, histopathology, suitable treatment and recurrence of nephrogenic adenoma, a rare, benign lesion of the urinary tract mucosa. PATIENTS AND METHODS: The records of 31 patients with nephrogenic adenoma diagnosed at the Sahlgrenska University Hospital between 1980 and 1996 were reviewed to determine the symptomatology, imaging investigations, endoscopic presentation, clinical outcome after resection and the frequency of recurrence. RESULTS: The lesions were found in the urinary bladder, bulbar urethra, urethral diverticula and the prostatic urethra. Eight patients presented with haematuria, 13 complained of urinary frequency and bladder pain and in 12 patients without subjective symptoms from the urinary tract, the lesion was found accidentally. Twenty-seven of the patients had a history of previous urothelial trauma, either by instrumentation or inflammation. Seventeen of the lesions were polypoid at endoscopy, the remainder being flat except for two cases, in which they were not noted because they were concealed in another lesion to be resected. Seven patients had one or more recurrences. All patients with symptoms responded well to transurethral resection. CONCLUSION: Nephrogenic adenoma mimics tumour or chronic cystitis and it is rarely suspected on clinical grounds; instead, the diagnosis is almost always histological. This study supports the view that nephrogenic adenoma may represent a metaplastic response to trauma of the urothelium and that transurethral resection provides a good method of relieving the symptoms in symptomatic nephrogenic adenoma. PMID- 9352689 TI - Flow cytometric analysis of tumour-infiltrating lymphocytes in patients with renal cell carcinoma. AB - OBJECTIVE: To determine the immunophenotype of tumour-infiltrating lymphocytes (TIL) and peripheral blood lymphocytes (PBL) isolated from patients with renal cell carcinoma (RCC) and to analyse the correlations between the quantity of analysed cell subsets and the progression of the disease. PATIENTS AND METHODS: PBL and TIL samples were obtained from 23 patients with RCC at different stages of disease. The immunophenotype of PBL and TIL was measured, and the TNM stage, tumour size, cellular type, histological grade, lymphocytic infiltration and performance status assessed. RESULTS: The predominant mononuclear cells infiltrating the tumour, in all patients, were T lymphocytes (CD3+ median 66.9%, CD8+ median 34.6%, CD4+ median 26.7%). The cells possessing gamma/delta type T cell receptor accounted for a small fraction of the T cells in PBL and TIL (median 5.6% and 3.7%). Tumour-infiltrating T lymphocytes had a significantly higher percentage of cells expressing human leucocyte antigen (HLA) DR (median 30.9%) and CD25 (median 6.2%) antigens than the equivalent populations in peripheral blood from the same patient group (P < 0.001). The degree of T cell activation appeared to negatively correlate with the tumour stage (K = -0.3, P = 0.04). The percentage of natural killer (NK) cells among TIL (median 15.4%) did not reflect the value in PBL. The percentage of B cells in TIL was slightly lower than in PBL and accounted for 5.0% of cells. There was no relationship between the degree of lymphocytic infiltration and either tumour stage or grade but there appeared to be a positive correlation between the intensity of lymphocytic infiltration and the percentage of CD4+ cells in TIL (K = 0.5, P = 0.001). Moreover, the composition of TIL depended on tumour grade, which positively correlated with the percentage of CD8+ cells (K = 0.4, P = 0.005) and negatively with the percentage of NK cells (K = -0.5, P = 0.005). There was an inverse correlation with the percentage of gamma/delta T cells in PBL and the TIL concentration (K = -0.3, P < 0.05). CONCLUSIONS: The TIL immunophenotype is different from PBL and is influenced by the histological grade of the tumour. The activation of TIL and its relationship with tumour progression suggests that they might be sensitized and activated by tumour cells. PMID- 9352690 TI - Early genetic and cellular responses in the smooth muscle layer of obstructed ureters in a rat model of obstructive uropathy. AB - OBJECTIVE: To investigate the early genetic and cellular responses in the smooth muscle layer of completely obstructed ureters, and to determine whether myocytes proliferate (hyperplasia) in the ureters during the early stage of obstructive uropathy. MATERIALS AND METHODS: The study comprised 35 female Sprague-Dawley rats which had undergone unilateral ligation of their ureters. After ureteric ligation, five rats each were killed and examined at 0.5, 1, 2, 4, 8, 16 and 24 h after ligation. The proximal portion of the ureters was prepared for light and electron microscopy. The expression of c-Fos, c-Jun, c-Myc and Ki-67 antigen was assessed immunohistochemically. RESULTS: c-Fos and c-Jun were detected 2 h after ligation and the expression of these two proteins reached a maximum after 4 h, becoming undetectable 16 h after ligation. The expressions of c-Fos and c-Jun were strongly correlated (r = 0.9854, P < 0.001). The expressions were of c-Myc and Ki-67 antigen was not detected within 24 h after ureteric ligation. The amount of rough endoplasmic reticulum (rER) in the ligated ureters increased soon after complete ligation and the increase continued throughout the period of ureteric obstruction (r = 0.9699, P < 0.001). The change in rER was also significantly correlated with the expression of c-Fos and c-Jun within 8 h after ligation. CONCLUSION: The expression of c-Fos and c-Jun, but not c-Myc, might contribute to the hypertrophy of the ureteric smooth muscle layer during the course of complete ureteric obstruction. There is no hyperplasia in ureteric smooth muscle in the early stages of obstructive uropathy. PMID- 9352691 TI - A prospective, randomized trial comparing continuous bladder drainage with catheterization at abdominal hysterectomy. AB - OBJECTIVE: To compare the infection rate and post-operative morbidity between in dwelling urinary catheterization and 'in-out' catheterization at the time of routine total abdominal hysterectomy. PATIENTS AND METHODS: The study comprised 100 patients who were blindly randomized to have either an indwelling Foley catheter or an 'in-out' catheterization at the time of surgery. Follow-up data on the retention of urine, urinary symptoms and infection were obtained. RESULTS: Of the 95 patients with complete data, 36% of those undergoing in-out catheterization had urinary retention after operation, requiring bladder emptying, compared with 4% of those receiving an indwelling catheter (P < 0.001). In addition, 29% of the catheterized group had urinary tract bacteriuria compared with 13% of the uncatheterized group (P < 0.025). CONCLUSION: This randomized controlled trial showed that in-out urinary catheterization at the time of routine abdominal hysterectomy was associated with a significantly higher incidence of post-operative urinary retention compared with in-dwelling catheterization, and may have implications for long-term bladder function. PMID- 9352693 TI - Prevalence of enuresis and other bladder symptoms in patients with active Graves' disease. AB - OBJECTIVE: To determine the prevalence of bladder symptoms in patients with Graves' disease and assess their abnormalities on urodynamic studies. PATIENTS AND METHODS: The study comprised 30 consecutive patients (13 female, 17 male, mean age 31 years, SD 10) with active Graves' disease. Before and after attaining euthyroidism with carbimazole, their bladder symptoms were assessed using a questionnaire adapted from the American Urologic Association voiding symptom score, the serum levels of total triiodo-L-thyronine (T3), serum thyroxine (T4) and thyroid-stimulating hormone (TSH) were measured by radioimmunoassay, and uroflowmetry, cystometry and perineal muscle electromyography were performed. RESULTS: Twelve of the 30 patients (40%) had the onset of bladder symptoms 1-6 months after the onset of symptoms of Graves' disease; four of the 12 patients had enuresis. Urodynamic studies were possible in five patients and showed reduced flow rates in all, a significant post-void residual volume in four, and enlarged bladder capacity and increased perineal muscle electromyographic activity during the voiding phase in three. Voiding disturbances and urodynamic abnormalities resolved after attaining euthyroidism. CONCLUSION: Significant bladder symptoms can occur in about 40% of patients with active Graves' disease but resolve on euthyroidism; in some patients, urodynamic studies show significant abnormalities which are also reversible. PMID- 9352692 TI - Willingness to pay for reduced incontinence symptoms. AB - OBJECTIVE: To measure the willingness to pay for a reduction in the number of micturitions and urinary leakages for patients with urge incontinence. PATIENTS AND METHODS: A self-administered questionnaire with a binary willingness-to-pay question was administered to 541 patients in Sweden with urge or mixed incontinence; 461 questionnaires were returned. The reduction in micturitions and urinary leakages valued in the willingness-to-pay question was varied randomly between 25% and 50% in two different subsamples. Information was also collected about the number of micturitions and urinary leakage, health-related quality of life and socio-economic characteristics of the patients in the study. RESULTS: Quality of life was significantly related to the severity of the symptoms and was worse than that of the sex- and age-matched general Swedish population. The median (mean) willingness to pay per month was 240 (530) Swedish krona (SEK, 1 Pound = SEK 11.50) for a 25% reduction in micturitions and leakages and SEK 470 (1030) for a 50% reduction in micturitions and leakages. As hypothesized, the willingness to pay was significantly related to the size of the reduction in micturitions and leakages, the initial number of micturitions and leakages, and income. CONCLUSIONS: Patients with incontinence problems are willing to pay substantial amounts for a reduction in the number of micturitions and leakages. PMID- 9352694 TI - Pitfalls of measuring residual urine volume in orthotopic neobladders. AB - OBJECTIVE: To investigate the pitfalls of measuring postvoid residual urine volume (PVR) in patients with orthotopic neobladders, in an out-patient clinic. PATIENTS AND METHODS: Fifteen patients with orthotopic neobladders were enrolled in the study; five patients had a colonic neobladder, five an ileocolic neobladder and five a sigmoid neobladder. The PVR was compared after measuring by catheterization under fluoroscopic monitoring after normal or forced voiding. RESULTS: The PVR after forced voiding was always less than that after natural voiding, regardless of the type of neobladder. CONCLUSIONS: Patients with orthotopic neobladders may have a higher PVR after voiding naturally at home than when the PVR measured in hospital. PMID- 9352695 TI - Transurethral prostatectomy: a prospective randomized study of conventional resection and electrovaporization in benign prostatic hyperplasia. AB - OBJECTIVE: To compare transurethral electrovaporization of the prostate (TUVP) with conventional transurethral resection of the prostate (TURP) in the treatment of men with benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: Seventy consecutive patients with symptomatic BPH and a prostate size of < 60 g were prospectively randomized between equal treatment groups; one group underwent standard TURP and the other TUVP. Patients were assessed at baseline and 1, 3, 6 and 12 months after treatment, giving a mean (SD) duration of follow-up of 14.4 (1.9) months (range 12-17). Variables evaluated included the duration of operation, catheterization and hospital stay, and changes in blood levels of haemoglobin, haematocrit and sodium 1 h after the operation. The American Urologic Association (AUA)-7 symptom score, peak urinary flow rate (Qmax), post voiding residual urine volume (PVR) and sexual function were also evaluated during the follow-up. RESULTS: Patients of both groups were balanced for the different baseline variables. The mean (SD) operative duration of TUVP was 52 (12.5) min, significantly longer than that of TURP, at 39.7 (8.8) min (P < 0.001). One hour after TURP, patients had significantly lower levels of haemoglobin, haematocrit and Na. The mean (SD) duration of catheterization after TURP was 2 (0.8) days, significantly more than after TUVP, at 1.1 (0.4) days (P < 0.001). The mean (SD) hospital stay was 2.5 (1) days after TURP and 1.5 (0.7) after TUVP (P < 0.001). Compared with baseline values, the AUA-7 symptom score, Qmax and PVR improved significantly in both groups at all intervals of follow-up and there were no significant differences between the groups during the follow up. None of 15 potent men undergoing TURP and two of 18 potent men undergoing TUVP complained of impotence during the follow-up. CONCLUSIONS: TUVP is as effective as TURP in the treatment of BPH in men with a prostate size of < 60 g. TUVP has the advantages of less blood loss, less absorption of irrigant and a shorter hospital stay, but it had a significantly longer operative duration. PMID- 9352696 TI - Electrovaporization of the prostate in patients with benign prostatic enlargement. AB - OBJECTIVE: To determine the effectiveness and safety of transurethral electrovaporization in the treatment of benign prostatic enlargement (BPE). PATIENTS AND METHODS: The study comprised 91 patients (median age 65 years) with BPE (median prostate volume 61 mL). Patients were assessed with a general and urological history, the International prostate symptom score (IPSS), urinary tract and prostatic ultrasonography, uroflowmetry and biopsy of the prostate. An electrosurgical generator (cutting at 200 W and coagulating at 70 W) with grooved electrodes was used to vaporize the prostate. The variables assessed before and after treatment were the IPSS, quality-of-life score, uroflowmetry, complications, and the duration of hospitalization. RESULTS: The mean operative duration was 45 min, blood loss was negligible and the mean duration of catheterization was 24 h. Recatheterization was necessary in 5.5% of patients. The median values before and after treatment were: IPSS, 19 and 5; QOL score, 4 and 2; and maximum urinary flow rate 8.3 and 22.1 mL/s (all P < 0.001). In nearly all cases, the Siroky nomogram showed that patients became unobstructed. CONCLUSIONS: Electrovaporization of the prostate has many advantages over other techniques used in the treatment of BPE, including transurethral resection (TURP). It is cheaper than laser-assisted prostatectomy in particular and is also simpler to perform. Higher-risk patients can be treated, it uses existing electrosurgical equipment with minor modifications and only the electrodes, which are inexpensive, differ from TUR loops; the TURP resectoscope can be used and there is almost no need for blood transfusion. Intra- and post-operative morbidity is very low, recatheterization rare, the hospital stay brief and the results equal to or better than those obtained with TURP. Convalescence is generally uneventful and rapid because haematuria is rare and, when present, mild. Patients can return to work earlier than after TURP. Although a promising technique, only a longer follow-up will allow firm conclusions about its suitability. PMID- 9352697 TI - Safety and efficacy of transurethral needle ablation of the prostate for symptomatic outlet obstruction. AB - OBJECTIVES: To examine, in an observational study, the safety and efficacy of transurethral needle ablation (TUNA) of the prostate as a treatment for symptomatic benign prostatic enlargement. PATIENTS AND METHODS: This prospective study included 71 symptomatic men with unequivocal obstruction on pressure-flow urodynamics. The variables measured at baseline and up to 12 months after treatment included the American Urological Association (AUA)-7 symptom index and an added quality-of-life question, the AUA BPH-Impact Index, a sexual function score, transrectal ultrasonography of the prostate, a frequency-volume chart, free-flow uroflowmetry, post-void residual urine volume (PVR) and pressure-flow urodynamics. Transurethral resection of the prostate (TURP) was offered if the symptoms failed to resolve at any time during the follow-up period. TUNA was performed under local anaesthetic and sedation in 63 (89%) men and as a day-case procedure in 10 (14%). Five patients were on warfarin which was not discontinued. RESULTS: There were no serious treatment-related adverse events. Eight of the initial nine patients who were not routinely catheterized after treatment with TUNA developed acute urinary retention. Although some haematuria occurred in all patients, only one (1.4%) developed catheter blockage by clot. There were no problems with bleeding in those patients on warfarin at the time of treatment. The mean (95% confidence interval, CI) AUA-7 index fell from 23 (1.7) to 10.6 (1.8) (P < 0.001, Mann-Whitney U-test) at 12 months, 29 men (41%) had an AUA-7 index of < or = 7. The maximum (95% CI) urinary flow rate increased from 9.0 (0.8) to 11.3 (1.1) mL/s (P < 0.001) and this was accompanied by a small but significant reduction in PVR of 70 (14) mL to 35 (8) mL (P < 0.001 Mann-Whitney U test). There was a significant reduction in both maximal voiding pressure and detrusor pressure at peak flow at 3 months (Mann-Whitney U-test, both P < 0.001) and at 12 months (P < 0.001, Wilcoxon matched-pairs signed-ranks test). However, 78% of the 45 men undergoing repeat pressure-flow studies at 12 months were unequivocally obstructed according to the Abrams-Griffiths nomogram. The mean (95% CI) prostatic volume fell from 49.0 (4.8) mL at baseline to 40.8 (4.9) mL at 3 months, but this change was not statistically significant (P = 0.011, Mann Whitney U-test). Two men reported erectile dysfunction, one experienced ejaculatory problems and seven reported an improvement in erectile function after TUNA. During the study, 22 men (31%) underwent TURP. CONCLUSIONS: TUNA is a safe treatment which can be performed as an out-patient procedure under local anaesthesia and sedation in the vast majority of patients. There was no evidence of serious adverse events and no significant adverse effect on sexual function. The symptomatic improvement was sustained at 12 months in most (54%) patients, with modest improvements in peak flow rate, PVR and voiding pressures, indicating that TUNA may result in prolonged symptomatic improvement in a proportion of patients suffering from bladder outlet obstruction. A randomized controlled study against established therapies is now essential to clearly delineate its place in the management of such patients. PMID- 9352698 TI - A dose-ranging study of the efficacy and safety of tamsulosin, the first prostate selective alpha 1A-adrenoceptor antagonist, in patients with benign prostatic obstruction (symptomatic benign prostatic hyperplasia). AB - OBJECTIVE: To evaluate the efficacy and safety in a dose-ranging study of tamsulosin (once-daily) as a modified-release formulation compared with placebo in patients with lower urinary tract symptoms (LUTS) associated with benign prostatic obstruction (BPO), and to establish the optimum dosage for phase III clinical studies. PATIENTS AND METHODS: Of 169 patients with LUTS associated with BPO enrolled in a 3 week placebo run-in period, 126 were subsequently randomized to receive placebo (28), or 0.2 mg (35), 0.4 mg (30), or 0.6 mg (33) of tamsulosin once daily for 4 weeks. Free-flow and pressure-flow measurements, and modified Boyarsky symptom scores were used to determine efficacy. Safety was evaluated by monitoring adverse events and vital signs (including 8 h after the first dose), and by laboratory determinations. RESULTS: Tamsulosin 0.4 mg and 0.6 mg produced significantly greater improvements in maximum urinary flow rate (Qmax) (2.2 mL/s, 22.6%, and 1.8 mL/s, 20.2%, respectively) than did placebo ( 0.1 mL/s, -0.9%). The results from the pressure-flow studies confirmed the results for Qmax in the free flow studies, with optimum and significant effects for tamsulosin 0.4 mg. This also applied for detrusor pressure at maximum flow, which decreased by 26.6 cmH2O (-28.2%) on 0.4 mg tamsulosin whereas it increased by 4.9 cm H2O (5.7%) on placebo. The greatest reductions in total symptom score were obtained with tamsulosin 0.4 mg and 0.6 mg (4.1, -28.7%, and 4.4 points, 28.2%, respectively) compared with reductions of 3.4 (-20.1%) in the tamsulosin (0.2 mg) and 2.9 points (-17.7%) in the placebo groups. The difference in effects on total symptom score between treatment groups was not statistically significant, which can be attributed to the small sample size. Tamsulosin was well tolerated; at least one adverse event was reported by 29%, 23%, 27% and 36% of patients in the placebo and tamsulosin 0.2 mg, 0.4 mg and 0.6 mg groups, respectively. There were no apparent tamsulosin dose-dependent changes in vital signs from baseline to the end of 4 weeks of randomized treatment. Tamsulosin caused no statistically significantly greater changes in blood pressure than placebo during the initial 8 h after the first dose. There were no clinically significant changes in laboratory variables. CONCLUSION: Tamsulosin is well tolerated and effective in improving urinary flow and relieving LUTS associated with BPO. Optimal effects are achieved with tamsulosin 0.4 mg administered once daily. PMID- 9352699 TI - Comparison of tamsulosin with alfuzosin in the treatment of patients with lower urinary tract symptoms suggestive of bladder outlet obstruction (symptomatic benign prostatic hyperplasia). The European Tamsulosin Study Group. AB - OBJECTIVE: To compare the efficacy and tolerability of the alpha 1 A-subtype selective drug tamsulosin with the nonsubtype-selective agent alfuzosin in the treatment of patients with lower urinary tract symptoms (LUTS) suggestive of bladder outlet obstruction (BOO), often termed symptomatic benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: The study comprised 256 patients with benign prostatic enlargement and LUTS suggestive of BOO (symptomatic BPH) who received tamsulosin 0.4 mg once daily or alfuzosin 2.5 mg three times daily during 12 weeks of treatment. The response was assessed by measurements of maximum urinary flow rate (Qmax), a symptom score (Boyarsky) and blood pressure at regular intervals. RESULTS: Tamsulosin and alfuzosin produced comparable improvements in Qmax and total Boyarsky symptom score. Both treatments were well tolerated with respect to adverse events. Tamsulosin had no statistically significant effect on blood pressure compared with baseline but alfuzosin induced a significant reduction in both standing and supine blood pressure, compared with baseline (P < 0.05). CONCLUSION: Tamsulosin is the first adrenoceptor antagonist that is selective for the alpha 1 A-subtype; this specificity may explain its lack of effect on blood pressure compared with alfuzosin, an agent that is not receptor subtype specific. Moreover, this finding may partly explain why tamsulosin, in contrast to other currently available alpha 1-adrenoceptor antagonists, can be administered without dose titration. Another advantage compared with alfuzosin (and prazosin) is the once-daily dosing regimen of tamsulosin. PMID- 9352701 TI - Morphometric studies of intra-prostatic volume relationships in localized prostatic cancer. AB - OBJECTIVES: To further characterize patterns of tumour growth and the distribution of markers for the aggressiveness of prostate cancer by assessing the relationships among the volume of the 'index' tumour and that of the remaining foci, with pathological (pT) stage, histological grade and DNA ploidy, and with the amount of low- and high-grade prostatic intraepithelial neoplasia (PIN). MATERIALS AND METHODS: Eighty-eight step-sectioned total prostatectomy specimens were analysed. The Gleason score, tumour stage and DNA ploidy (by flow cytometry) of multiple samples were determined. Tumour and PIN areas were outlined and their volumes estimated by computerized planimetry. RESULTS: The pT stage, Gleason sum and DNA nondiploidy increased, and PIN volumes decreased, with increasing volume of the index tumour focus (P < 0.01), but did not differ significantly between uni- and multifocal tumours. However, PIN volumes were significantly larger in multifocal cases with an index tumour volume of > 3 mL than in unifocal tumours > 3 mL (P < 0.05). Small volume, unifocal tumours had little PIN. The most malignant features of each case were always represented in the index tumour but not generally in the remaining foci. CONCLUSIONS: The volume distribution, related to multicentricity and its concomitant PIN volumes, indicates that large index tumours, uni- or multifocal, of medium or high grade, are associated with low PIN volumes. However, multifocal medium- and high-grade tumours with small index tumour volumes have higher PIN volumes. Small, single tumours are of low-grade and may represent the slowly progressing cancers possibly resembling those found in autopsy studies. PMID- 9352700 TI - Clinical comparison of selective and non-selective alpha 1A-adrenoreceptor antagonists in benign prostatic hyperplasia: studies on tamsulosin in a fixed dose and terazosin in increasing doses. AB - OBJECTIVE: To compare the efficacy and safety of a fixed dose (0.2 mg) of tamsulosin, a selective alpha 1A-adrenoreceptor antagonist, with an increasing dose (1-5 mg) of terazosin, a non-selective antagonist, in the treatment of urinary outflow obstruction associated with benign prostatic hyperplasia (BPH) in Korean patients. PATIENTS AND METHODS: The study comprised a single-blind and randomized design with tamsulosin or terazosin taken once daily for 8 weeks. A total of 98 patients was enrolled, with 72 patients included in the analyses after 4 and 8 weeks. The primary variables assessed were changes in the maximum urinary flow rate Qmax and the total International Prostate Symptom Score (IPSS), with the post-void residual urine volume, 'obstructive' and 'irritative' questions in the IPSS, and the investigators' global assessment of efficacy also determined. The number of patients with a clinically significant response to treatment with tamsulosin or terazosin was determined and defined as those with > 20% improvement from the baseline Qmax or > 20% decrease in total IPSS. Adverse reactions possibly or probably related to study medication were recorded throughout the treatment period. RESULTS: Both tamsulosin and terazosin produced similar significant improvements in subjective and objective symptoms of urinary outflow obstruction (P > 0.05). Systolic and diastolic (standing) blood pressures decreased significantly in patients treated with terazosin (P < 0.05). The adverse reactions, most frequently dry mouth and dizziness which were usually mild and transient, were significantly higher in patients on terazosin (18 patients, versus one on tamsulosin, P < 0.001). The changes led to discontinuation of therapy in two patients on terazosin. CONCLUSION: Tamsulosin was as effective as terazosin in treating urinary outflow obstruction associated with BPH, but had a markedly better safety profile. PMID- 9352702 TI - Free and total prostate-specific antigen serum concentrations do not help to detect prostate cancer in patients with urinary outlet obstruction. AB - OBJECTIVES: To determine whether different molecular forms of prostate-specific antigen (PSA) obtained before transurethral resection of the prostate (TURP) indicate the presence of prostate cancer. PATIENTS AND METHODS: The free, total and free-to-total PSA levels were measured in 261 patients scheduled for TURP, 20 of whom had known prostate cancer. The tissue histology was compared with the PSA levels and the patients were followed for 5 years. RESULTS: Prostate cancer was detected in 23 of the patients (9%) who were thought to have benign disease. Normal ranges for the distribution of the PSA levels were established based on the patients with a benign histology, but these ranges did not detect most of the unknown cancers. The sensitivity of the total PSA test in detecting cancer was 38% and the specificity 90%. The discrimination was no better when considering the free fraction or the free-to-total PSA level. However, none of the 14 patients whose cancer was missed showed general progression of the disease during the 5-year follow-up and only one died from prostate cancer. In contrast, eight of the 20 patients with a known prostatic malignancy showed general progression, and six died from the disease. CONCLUSION: PSA testing of patients with outlet obstruction often failed to detect prostate cancer, but the prognosis was moderately good in those patients in whom it was missed. PMID- 9352703 TI - Treatment of high-risk patients with subvesical obstruction from advanced prostatic carcinoma using a thermosensitive mesh stent. AB - OBJECTIVE: To evaluate the results obtained using a permanent prostatic stent system (Memotherm, Bard/ Angiomed, Karlsruhe, Germany) in high-risk patients with advanced prostatic carcinoma and subvesical obstruction. PATIENTS AND METHODS: The study included 35 patients (mean age 75.3 years, range 53-89) with advanced prostatic carcinoma and persistent subvesical obstruction despite androgen ablation. Because of serious concurrent diseases, 49% of these patients were classified as American Society of Anesthesiologists (ASA) grade 3 and 51% as ASA grade 4. The patients were treated using the Memotherm stent, a thermosensitive Nitinol mesh stent. The outcome was assessed by measuring voiding variables, a symptom score and as the incidence of complications. RESULTS: After inserting the stent, 33 (94%) of the patients were able to void spontaneously and there was a statistically significant improvement in the voiding variables. These results remained unchanged over a mean (range) follow-up of 15.2 months (3-38). There were no serious complications arising from the insertion of the stent. CONCLUSION: For high-risk patients with subvesical obstruction caused by prostatic carcinoma, the insertion of a permanent metal stent system offers a useful alternative treatment to transurethral resection. PMID- 9352704 TI - Diverticula of the female urethra. AB - OBJECTIVE: To determine the optimum procedure for the diagnosis and therapy of diveticula of the female urethra. PATIENTS AND METHODS: The study included 18 patients with urethral diverticula treated at the Cristo Re Hospital in Rome between 1987 and 1995. Most of the patients were suffering from cystitis (eight), dysuria (seven) and recurring urinary infections (11). Less frequently, more specific symptoms were present such as post-voiding dribbling (two) and anterior vaginal mass (three). The pre-operative evaluation included a history, physical examination, voiding and positive pressure voiding cysto-urethrography (VCUG) and urodynamic tests. A 'typical' surgical excision of the diverticula was carried out in all cases. Surgical excision was combined with cystopexis (Raz operation) in four patients with urinary stress incontinence and three with detrusor instability were treated postoperatively with anticholinergics for 3 months. The outcome was evaluated by a physical examination and urodynamic tests at 3, 6 and 12 months postoperatively; the mean (range) follow-up was 34 (2-80) months. RESULTS: All the urethral diverticula were in the distal two-thirds of the urethra, along the posterolateral wall. The VCUG was sufficient for diagnosis in eight patients while the other 10 required a positive-pressure VCUG. Fifteen patients were evaluated; complications included a urinary tract infection for 2 months in four patients and stress incontinence for 2 months in two. There were no recurrences or urethrovaginal fistulae. CONCLUSIONS: Diverticula in the female urethra are difficult to diagnose because the symptoms can be misleading; the positive-pressure VCUG is useful in doubtful cases. However, a detailed history and physical examination are mandatory. PMID- 9352705 TI - Male subfertility: diagnostic and therapeutic advances. PMID- 9352706 TI - Retrograde embolization and causes of failure in the primary treatment of varicocele. AB - OBJECTIVE: To assess retrograde embolization for the treatment of varicocele and to examine the causes of surgical and radiological treatment failure. PATIENTS AND METHODS: Of 154 patients with clinical varicocele associated with subfertility or symptoms who were treated, 100 underwent surgical high ligation, retrograde embolization under fluoroscopic control was attempted in 84 and 30 had both forms of treatment. Venographic findings were defined in those patients for whom embolization proved impossible and in those in whom prior high ligation had failed. Among subfertile patients, 64 had semen analyses before and at least 3 months after the procedure available for comparison. Those patients undergoing both radiological and surgical procedures were sent questionnaires to evaluate their experience. RESULTS: Retrograde embolization was technically successful in 68 (81%) of the 84 patients. Two early failures were associated with venous spasm provoked by technical inexperience, while difficulties in the remainder were caused by anomalous venous anatomy. In patients who had recurrent varicocele after previous ligation, venography showed incomplete ligation of collateral channels; 14 of 18 patients were successfully re-treated by embolization. The sperm concentration improved significantly in 83% of patients undergoing embolization and in 63% of those surgically ligated. Patients who underwent both procedures expressed a strong preference for embolization. CONCLUSION: In centres where there is a skilled interventional radiologist, embolization is an effective alternative to surgical ligation of varicocele. Carried out under local anaesthesia as an out-patient procedure, it is cost-effective, associated with minimal morbidity and most patients are able to return to normal daily activities immediately. PMID- 9352707 TI - A single-centre observational study of surgery and late malignant events after chemotherapy for germ cell cancer. AB - OBJECTIVE: To review the impact of surgical staging after treatment on the late malignant events in an unselected group of patients treated with chemotherapy for germ cell cancer of the testis over the last 16 years. PATIENTS AND METHODS: The study comprised 256 patients treated between 1978 and 1994 who were reviewed for late relapse and development of second germ cell and non-germ cell cancer. RESULTS: At diagnosis, 142 patients had clinical stage 2, 30 stage 3 and 84 stage 4 disease; 57 patients relapsed within 20 months of treatment, while late germ cell cancer relapses (> or = 24 months after treatment) occurred in six patients. Of patients relapsing early or late, 42% and 33%, respectively, received surgery after treatment. Only two of those relapsing late remain progression-free with further treatment. Four patients developed germ cell cancer in the contralateral testis, while six developed second non-germ cell cancers. CONCLUSION: Late events occurred in 6.2% of 256 patients in this series, from 29 to 141 months after treatment. Given that the late relapse rate of six of 256 (2.3%) is less than the incidence of mature teratoma at routine retroperitoneal lymph node dissection, more patients may eventually relapse. These results suggest that there might be a case to evaluate the use of ultrasonographic surveillance of the retroperitoneum and testis at 5, 10 and 20 years, in addition to extending routine surveillance. PMID- 9352708 TI - Does necrosis on frozen-section analysis of a mass after chemotherapy justify a limited retroperitoneal resection in patients with advanced testis cancer? AB - OBJECTIVE: To evaluate morbidity and relapse in patients with advanced testis cancer who underwent a post-chemotherapy resection of a residual mass and a limited retroperitoneal lymph node dissection. PATIENTS AND METHODS: A total of 62 patients underwent complete resection of retroperitoneal masses after chemotherapy and a limited lymph node dissection if frozen sections of the mass showed necrosis (37 patients) or a bilateral dissection if the frozen section indicated viable germ cell tumour or teratoma (25 patients). RESULTS: With a median follow-up of 6 years, 14 (23%) patients relapsed, but only one within the retroperitoneum (teratoma) after a limited lymphadenectomy. There was a concordance of 89% between the frozen section of the post-chemotherapy mass and the permanent-section histological diagnosis of the entire lymphadenectomy specimen. Of the 37 patients whose masses showed necrosis on frozen section, three had viable germ cell tumour and one had teratoma on final histology. In all four false-negative cases, residual tumour was confined to the resected mass. Six patients (10%) had surgical complications (one after limited and five after bilateral lymph node dissection). CONCLUSION: The surgical resection of all residual masses after chemotherapy, followed by limited retroperitoneal lymphadenectomy if frozen-section analysis shows necrosis, is a safe approach in selected patients with advanced testicular cancer. PMID- 9352709 TI - Frequency-volume chart data from incontinent children. AB - OBJECTIVE: To establish the mean and standard deviation about the mean for voiding variables of incontinent children aged 6-11 years as measured on a frequency-volume chart (FVC), and to determine the effect of type of incontinence, gender and age on these values. PATIENTS AND METHODS: All children attending two continence clinics over a 3-year period completed a FVC as a routine part of their assessment. Voided volume and voiding interval data were collected from these charts. The mean maximum, mean minimum and overall mean voided volume and voiding interval were established for the whole group and then for each age, gender and type of incontinence. RESULTS: The voiding patterns of incontinent children were very variable and thus the standard deviation for each voiding parameter was large. Multivariate analysis showed that the only variable that affected any of these apparent storage parameters was the child's age; gender and type of incontinence did not influence bladder storage patterns. Children with day-time incontinence did not have smaller voided volumes than those with nocturnal enuresis. CONCLUSION: Both the mean and the standard deviation about the mean of all voided volumes varied widely amongst incontinent children. Only age appeared to influence trends in voided volumes. Any clinical investigation using the FVC in children should consider the high standard deviation when calculating sample size. PMID- 9352710 TI - Histology of the upper pole in complete urinary duplication--does it affect surgical management? AB - OBJECTIVE: To review the histological change in the upper pole of excised duplex kidneys and assess whether ante-natal diagnosis might predispose to more conservative surgical management of this abnormality. PATIENTS AND METHODS: Fifty consecutive patients undergoing upper pole hemi-nephroureterectomy for ectopic ureter or ectopic ureterocele between 1980 and 1992 had their histology reviewed and assessed for dysplastic, inflammatory and obstructive change. RESULTS: Segmental scarring and chronic and acute inflammatory change occurred consistently and the degree of inflammation seemed unaffected by antenatal diagnosis. Dysplasia was seen in 70% of patients with ureterocele and in only 30% of those with ectopic ureter. One patient had normal histology. CONCLUSION: There was no evidence of reversible histological change in patients with ectopic ureter and ectopic ureterocele who were diagnosed ante-natally. Preservation of the upper pole of the kidney does not seem justified in the light of the histological evidence. PMID- 9352711 TI - Retrograde cerebral perfusion and hypothermic circulatory arrest to remove intracardiac renal tumour thrombi. PMID- 9352712 TI - Seromuscular myotomy to help eversion of an urostomy. PMID- 9352713 TI - Lactate dehydrogenase as a tumour marker in adult Wilm's tumour. PMID- 9352714 TI - Normal umbilicus and infra-umbilical abdominal wall in bladder exstrophy. PMID- 9352715 TI - Extramammary Paget's disease of the penis is associated with long-standing transitional cell carcinoma and radiotherapy. PMID- 9352716 TI - Indwelling catheter causing perforation of the bladder. PMID- 9352717 TI - Paraganglia as an unusual mimic of carcinoma in the prostate. PMID- 9352718 TI - Hibernoma in the scrotum. PMID- 9352719 TI - Urological complication of pelvic fracture treated by an external fixator. PMID- 9352720 TI - Methicillin-resistant Staphylococcus aureus cleared using a spiral thermoexpandable urethral stent. PMID- 9352721 TI - Rectus sheath haematoma resulting from a subcutaneous injection with goserelin. PMID- 9352722 TI - Recurrent intra-vesical foreign bodies. PMID- 9352723 TI - Unsolved problems. PMID- 9352724 TI - Neuropsychological, intellectual, and behavioral findings in patients with centrotemporal spikes with and without seizures. AB - Forty children (23 boys, 17 girls) with centrotemporal spikes (rolandic focus) with and without seizures (mean age 8.4 years +/- 4.8 SD), and 40 healthy controls matched for age, sex, and socioeconomic status were assessed for their neuropsychological, intellectual, and behavioral outcome. Compared with the controls, patients were significantly impaired in their IQ, visual perception, short-term memory, in their psychiatric status and in some subtests in a fine motor performance task. No significant differences could be computed for a simple finger-motor speed exercise or a linguistic performance test. In patients, deficits in IQ were significantly correlated with frequency of spikes in the EEG, but not with frequency of seizures, lateralization of the rolandic focus, or time since rolandic focus was diagnosed. It was concluded that a rolandic focus is not as benign as once thought. PMID- 9352725 TI - Status epilepticus in children: aetiology, treatment, and outcome. AB - This retrospective study includes 65 children treated for status epilepticus at Tampere University Hospital in Finland. Aetiology of the condition, effectiveness of the treatment protocol, including short barbiturate anaesthesia to prevent prolonged status epilepticus episodes, and neurological outcome were evaluated. Symptomatic aetiology was present in 40% of status epilepticus episodes, and 37% of episodes were induced by fever. Neurological sequelae secondary to status epilepticus were identified in 15% of the cases and subsequent epilepsy in 23% during the mean follow-up time of 3.6 years. There were no status epilepticus related deaths. The cut-off point of status epilepticus duration for significant risk for permanent neurological sequelae was 2 hours. Our treatment protocol, including short barbiturate anaesthesia in refractory cases, was able to abort status epilepticus in less than 2 hours in 75% of cases. We conclude that early and prompt use of barbiturate anaesthesia should be encouraged, and may explain our low morbidity figures. PMID- 9352726 TI - Epilepsy in patients with cerebral palsy. AB - The incidence of epilepsy in 323 patients with cerebral palsy (CP) was 41.8%. Almost half of the patients with spastic tetraplegia and hemiplegia had epilepsy. The incidence was lower in patients with spastic diplegia. No sex differences were observed. Partial seizures were by far the most common form of epilepsy in spastic hemiplegia, while generalized tonic-clonic episodes predominated in all other forms of CP. A very high incidence of West syndrome was observed in patients with spastic tetraplegia. Most of the patients with spastic tetraplegia had their first seizure in the first year of life. In patients with spastic hemiplegia the onset of epilepsy was often delayed for several years. A high rate of polytherapy was recorded, but two-thirds of the patients remained seizure-free for long periods. In just over one-fifth of the patients successful withdrawal of medication was achieved. PMID- 9352727 TI - A comparison of intensive neurodevelopmental therapy plus casting and a regular occupational therapy program for children with cerebral palsy. AB - The purpose of this research was to evaluate the combined effect of intensive neurodevelopmental therapy (NDT) and casting in improving hand function, quality of upper-extremity movement and range of motion in children aged between 18 months and 4 years with cerebral palsy (CP). A randomized crossover design was used to evaluate the difference between intensive NDT plus casting and a less intensive regular occupational therapy (OT) program. Blinded assessments of hand function, quality of upper-extremity movement, and parents' perception of hand function performance were carried out at baseline, 4 months (end of first intervention period), 6 months (after a 2-month 'washout' period), and 10 months (end of second intervention period). Analysis of the outcomes revealed no significant differences in hand function, quality of upper-extremity movement, or parents' perception of hand-function performance between the two treatment groups intensive NDT plus casting or regular OT programs. There does not appear to be any beneficial effect of an increased amount of therapy for the children in this study. PMID- 9352728 TI - Oxygen cost, walking speed, and perceived exertion in children with cerebral palsy when walking with anterior and posterior walkers. AB - Walkers with the frame positioned behind the child have been advocated recently, mainly because they may allow a more upright and therefore more normal ambulation and perhaps encourage favourable neuromuscular development. The purpose of this study was to estimate and compare speed, energy cost, and perceived exertion during walking with an anterior walker and a posterior walker. Ten children with spastic diplegia, average age 11 years, who were familiar with both types of walker participated in the study. Spasticity was measured according to the modified Ashworth scale. Oxygen cost was determined by the argon-diluted method using a mixing box mounted on a backpack, and the perceived exertion was graded. The results of the study showed that there are no differences in the measured variables in walking between the anterior and the posterior walker in children familiar with both walkers and also that most of the children preferred the posterior walker. PMID- 9352729 TI - Determinants of nocturnal enuresis in England and Scotland in the '90s. AB - The aim of this study was to assess whether changes have occurred in the determinants of nocturnal enuresis in Scotland and England in comparison with previous studies. The study was based on 22 study areas from a representative English sample, 14 areas from a representative Scottish sample, and 20 areas from an English inner-city sample. A total of 14,674 subjects was included in the analysis from 16,835 eligible children in the age range 5 to 11 years. For the main analysis, an enuretic child was one who wet the bed at least once a week. As expected, the frequency of enuresis was higher in boys and decreased markedly with age in both sexes. Bedwetting was more frequent in: Afro-Caribbean children compared with white children in the representative samples (OR 1.72 95% CI 1.22 to 2.42); those whose mothers smoked at least 10 cigarettes at home compared with non-smokers (OR 1.58 95% CI 1.26 to 1.98); children who had disturbed sleep compared with those who slept well (OR 1.96 95% CI 1.53 to 2.51); those with mothers aged less than 20 years at the child's birth compared with mothers in the age range 25 to 34 (OR 1.63 95% CI 1.20 to 2.22); and in the second- or third born in the family in comparison with the first-born (OR 1.42 95% CI 1.17 to 1.72). Father's social class was associated with enuresis only in girls. Only 50% of the parents consulted a doctor for enuresis in their child. The percentage was even lower in Afro-Caribbean families (33%). Enuresis continues to be a highly prevalent problem and has not decreased over the last 45 years. We confirm that environmental factors are still important in the aetiology of enuresis. It is surprising that despite the availability of effective treatment only half of parents consult a doctor about the problem. PMID- 9352730 TI - Helicobacter pylori in an institution for disabled children in Hong Kong. AB - Anti-Helicobacter pylori antibodies were determined in 157 institutionalised Cantonese children, mean age 9.5 +/- 3.9 (SD) years, with profound neurodevelopmental disabilities. Eighty-seven (55.4%) were H. pylori seropositive compared with four of 50 (8%, P > 0.0002) of an age-matched control group, mean age 7.2 +/- 4.3 (SD) years. Eight of 15 seropositive children with a recent history of upper gastrointestinal bleeding underwent endoscopy and in all cases gastric infection with H.pylori was confirmed. Anthropometric data from institutionalised children revealed marked malnutrition but showed no significant difference between seropositive and seronegative children. Disabled children receiving long-term residential care in Hong Kong are confirmed to be at increased risk of H.pylori infection. PMID- 9352731 TI - Complex orofacial movements and the disappearance of cerebellar mutism: report of five cases. AB - A syndrome of mutism and subsequent dysarthria occurs frequently in children after resection of a cerebellar tumour. The role of orofacial and speech motor control in this syndrome has not been studied systematically. We examined simple and complex orofacial movements during the mute phase and shortly after the resumption of speech in five children with mutism and subsequent dysarthria. The recovery of complex orofacial movements coincided with the disappearance of the mutism. PMID- 9352732 TI - Boys with Asperger's disorder, exceptional verbal intelligence, tics, and clumsiness. AB - Five boys with both Asperger's disorder and Tourette syndrome, exceptional verbal intelligence, and clumsiness are reported. Each presented at early elementary school age with a prominent complaint of social difficulties with peers. History was notable for a flapping stereotypy and the neurological examination revealed motor and/or vocal tics and numerous motor soft signs. Highly specialized interests were characteristics. Language prosody and/or pragmatics was impaired. Despite exceptional verbal intelligence, the children were not, according to their teachers and parents, faring well either socially or academically. Motor difficulties, manifested psychometrically as a significant performance IQ disadvantage, interfered with school performance and social adjustment. Tics, although not noted by parents in the clinical history, compounded their social difficulties. Asperger's disorder in these highly verbal children overlaps with pervasive developmental disorder (PDD) on account of the socioemotional difficulties and stereotypies seen in both. Asperger's disorder and Tourette syndrome overlap in these children on account of the tics. Finally, Asperger's disorder and the right-hemisphere-based learning disorders overlap on account of the visuoperceptual and attentional deficits that can occur in both. PMID- 9352733 TI - Pure congenital Foix-Chavany-Marie syndrome. AB - Foix-Chavany-Marie syndrome (FCMS) is characterized by facio-linguo-masticatory diplegia in the absence of limb weakness. The most common cause is a cortical lesion resulting from a stroke but a congenital form has been reported. We present the case of a 53-year-old man who was admitted to hospital with worsening dysphagia which was know to have been present together with anarthria and facial palsy, since birth. He demonstrated features of FCMS with pseudobulbar palsy and unaffected reflexes and automatic responses. Cranial CT and MRI scans showed bilateral opercular lesions of CSF intensity in continuity with the lateral ventricles. We conclude that this case of static FCMS for over 50 years may represent a 'pure' form of congenital FCMS with motor symptomatology and unaccompanied by mental retardation or epilepsy. PMID- 9352735 TI - Postural support systems: their fabrication and functional use. PMID- 9352734 TI - Visual agnosia with bilateral temporo-occipital brain lesions in a child with autistic disorder: a case study. AB - A 2-year-old boy meeting the criteria for autistic disorder was diagnosed 2 years later with a visual agnosia characterised by a combination of certain aspects of associative and apperceptive agnosia. MRI then revealed a severe encephalomalacia of the right temporal lobe and bilateral temporo-occipital areas. This association is discussed in terms of a clinical and aetiological relation between autistic disorder and visual agnosia. PMID- 9352736 TI - Huntington disease and the related disorder, dentatorubral-pallidoluysian atrophy (DRPLA). PMID- 9352737 TI - Histoplasmosis. Experience during outbreaks in Indianapolis and review of the literature. AB - Histoplasmosis remains a common infection in endemic regions of North America and Latin America, causing a broad spectrum of clinical findings. Experience during recurrent outbreaks in Indianapolis has shown the importance of immunosuppressive conditions including the acquired immunodeficiency syndrome (AIDS) as a risk factor for disseminated disease and expanded our knowledge of the common clinical manifestations. Pericarditis, rheumatologic manifestations, esophageal compression, and sarcoidlike manifestations were found to be relatively common findings in histoplasmosis. These studies have established the useful role of serologic testing and have led to the discovery of antigen testing for diagnosis of histoplasmosis. This experience also has offered the opportunity to examine the outcome of treatment in persons with AIDS, contributing to studies that have found itraconazole to be an excellent alternative to amphotericin B in persons with mild or moderately severe infection. PMID- 9352738 TI - Renal vascular lesions in lupus nephritis. AB - We retrospectively studied the prevalence, histologic features, clinical correlations, and long-term outcome of the intrarenal vascular lesions of lupus nephritis (LN) in a series of 169 renal biopsies performed between 1980 and 1994 in 132 patients with systemic lupus erythematosus. The most common vascular lesions were nonspecific sclerotic changes, found in 37% of the biopsies (24% if only the cases with moderate to severe changes are considered). The other common vascular lesions were "immunoglobulin microvascular casts," found in 24% of the biopsies. Vasculitis and thrombotic microangiopathy were rare lesions and were seen in only 4 (2.4%) and 1 (0.6%) cases, respectively. Isolated sclerotic vascular changes were present in biopsies from older patients with a longer duration of LN, compared with the group with no vascular lesions, and were associated with a significantly higher prevalence of hypertension. Overall, however, the long-term renal and patient survival of this group did not differ significantly from that of the patients without vascular changes. Immunoglobulin microvascular casts (IMCs) ("lupus vasculopathy") were characterized by the presence of immunoglobulin deposition within the glomerular capillaries and small arterioles. In the present study we extensively investigated the morphologic and immunologic features of this lesion. The lesions were notable for the absence of endothelial or parietal vascular lesions and of fibrin, platelets, and leukocytes, which indicates that thrombosis is not involved in the vascular obstruction. According to our data immunoglobulin precipitation in the microvasculature seems to play a central role in the pathogenesis of this lesion, which is why we propose the term "immunoglobulin microvascular casts." In general, IMCs were associated with the most severe and active forms of diffuse proliferative lupus nephritis (World Health Organization [WHO] class IV). However our data show that, in contrast to previous studies, the long-term outcome of patients with IMCs is not worse than that of other patients with class IV LN. It may even be somewhat better, suggesting that this type of lesion may reverse with immunosuppressive therapy. In addition, we did not find any association between the presence of IMCs and the lupus anticoagulant, IgG anticardiolipin antibodies, or extrarenal vascular manifestations. Concerning vasculitis and thrombotic microangiopathy, our results confirm that their occurrence is quite rare in-lupus nephritis. The outcome of our 4 patients with vasculitis was not particularly poor, which could be related to early and/or aggressive treatment. Taken as a whole, our data confirm that the presence of active and severe forms of diffuse proliferative LN (WHO class IV) carries a worse prognosis compared with the other forms of LN. In our study, and in agreement with previous reports (23), the long term renal survival of patients with class IV LN was significantly worse than that of patients with other forms of LN, with a 10-year renal survival of 70% compared with 85%, respectively. However our data do not support the conclusions of some previous studies that the presence of intrarenal vascular lesions is a marker of poor renal prognosis in lupus nephritis. More precisely, our data show that the somewhat poorer renal outcome observed in patients with IMCs is related to the fact that in most cases these lesions are associated with class IV lupus nephritis, and not related to the presence of the vascular lesion per se. PMID- 9352739 TI - Manifestations resembling thrombotic microangiopathy in patients with advanced human immunodeficiency virus (HIV) disease in a cytomegalovirus prophylaxis trial (ACTG 204). PMID- 9352740 TI - Serum levels of G-CSF, IL-3, IL-6 and GM-CSF after a single intraperitoneal dose of rhG-CSF in lethally irradiated B6D2F1 mice. AB - The objective of this study was to assess the pharmacokinetics of rhG-CSF after a single intraperitoneal injection 2 h post-TBI in B6D2F1 lethally irradiated mice and to analyze the effect of rhG-CSF on the endogenous response of interleukin-3 (IL-3), interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in these animals. For comparison, these cytokine serum levels have also been measured in nonirradiated mice. The serum concentrations of rhG-CSF in irradiated mice were higher than in nonirradiated mice at all time points during the first 60 min after injection. Furthermore, rhG-CSF administration failed to induce detectable endogenous serum levels of IL-3, IL-6 and GM-CSF, at least in the 72-hour period after administration of the rhG-CSF. The radioprotective effect of rhG-CSF in lethally irradiated mice is not mediated by an increase in endogenous serum levels of these three cytokines. PMID- 9352741 TI - Hydroxyurea therapy in sickle cell anemia patients in Curacao, The Netherlands Antilles. AB - We have treated 9 patients with sickle cell anemia (SS) with hydroxyurea (HU). All 9 patients carried 4 alpha-globin genes and the beta s-globin haplotypes 19/19 (Benin/Benin), except for 1 who had haplotype 19 together with type 3 (Benin/Senegal). Six patients received HU for 10 months and were again treated with the drug for 5 months after an interval of 1 year. One patient was given HU for 22 consecutive months. A record was kept of hematological and biochemical data, Hb F and G gamma levels, as well as possible clinical complications. Our data show that HU generally improves the hematological and biochemical values and the level of Hb F, and reduces painful crises in some patients. However, although the clinical symptoms improved in some patients during HU therapy, the older patients did not observe any changes in their general condition; the same is the case for the patient with haplotype 19/3. One patient also experienced life threatening liver sequestration during treatment. We conclude that the selection of patients who may benefit from HU therapy needs further evaluation. PMID- 9352742 TI - Prognostic factors in elderly patients with non-Hodgkin's lymphoma treated with cyclophosphamide, vincristine, prednisone, bleomycin, Adriamycin, procarbazine (COP-BLAM) therapy. AB - Elderly patients with non-Hodgkin's lymphoma (NHL) were treated with cyclophosphamide, vincristine, prednisone, bleomycin, Adriamycin, procarbazine (COP-BLAM) therapy at our institution. Prognostic factors were analyzed in 62 patients with untreated NHL aged 65 years or older. Of these patients, 47 (75.8%) achieved a complete remission and 11 (17.7%) partial remission. The overall response rate was 93.5%, with a 5-year survival rate of 70%. Factors with prognostic significance included age, performance status, albumin, stage, B symptoms, and histologic type according to the International Working Formulation. Of the 62 patients, 26 were alive at the time of writing. Of the 36 patients who died, 30 died due to progression of NHL or treatment-related disorders, and 6 died from disease other than NHL. Some of the prognostic factors identified in these elderly patients are not included among the prognostic factors reported for younger patients, suggesting the need to individualize chemotherapy for NHLs of different types, which can be defined by prognostic factors. Prognostic factors other than age should be taken into account, particularly when doses and dosing intervals are determined. PMID- 9352743 TI - Use of high-dose chemotherapy plus granulocyte colony-stimulating factor for the salvage of refractory or resistant-relapse lymphoma patients without stem cell support. AB - The combination of cyclophosphamide (CY) and etoposide is synergistic, spares bone marrow stem cells and can be given repeatedly in high doses without stem cell support. Thirteen patients with non-Hodgkin's lymphoma (n = 8) or Hodgkin's disease (n = 5), received high-dose chemotherapy (HDC). Median age was 32 years (24-52). Male to female ratio was 10:3. All the patients were in advanced-stage. Karnofsky score prior to HDC was 60% (range 40-90). Six patients showed primary refractoriness and 7 had resistant relapse. HDC consisted of CY 1,500 mg/m2/day and etoposide 300 mg/m2/day, both for 4 days. rhG-CSF was started 24 h after the last dose of chemotherapy as a continuous intravenous infusion at a dose of 0.01 mg/kg/day and stopped when the leukocyte count reached 1 x 10(9)/1 on 3 consecutive days. Overall, 69% (9/13) of patients responded to HDC. Four achieved CR and 5 achieved PR. Two of the patients showed disease progression. The other 2 died during the early period of HDC. Neutrophil and platelet recovery after HDC were 8 (6-16) and 10 (4-14) days, respectively. The major nonhematological toxicities were nausea-vomiting (100%) and diarrhea (61%). The median follow-up was 204 (7-600) days. Two patients relapsed 48 and 185 days after HDC. Eight patients are still alive, 7 progression free. The progression-free survival is 220 (40-285) days. In conclusion, HDC + granulocyte colony-stimulating factor (G CSF), without stem cell support seems to be promising in refractory or resistant relapse lymphoma patients bringing the need for randomized studies to show the cost effectiveness of HDC + G-CSF compared to HDC + autologous stem cell support. PMID- 9352744 TI - Possible involvement of bone marrow stromal cells in agranulocytosis caused by vesnarinone treatment. AB - Vesnarinone, an oral therapeutic agent for cardiac failure, causes agranulocytosis as a side effect. To elucidate the mechanism of occurrence of the agranulocytosis, we examined the effect of vesnarinone on granulopoiesis using an in vitro human long-term bone marrow culture system. Addition of vesnarinone to the culture decreased the total number of hematopoietic cells, mainly composed of mature granulocytes and macrophages, but increased the number of granulocyte macrophage progenitor cells (CFU-GM) and CD33-CD34+ cells as compared with an untreated control. Differentiation of CFU-GM was induced by removing the agent from the culture medium, indicating that the effect of vesnarinone was reversible. The agent did not directly affect CFU-GM in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Furthermore, treatment of stromal cells with vesnarinone repressed the production of G, GM, M-CSF, suggesting that the agent may cause a hematopoietic disorder, agranulocytosis, through the impairment of stromal cell function. PMID- 9352745 TI - Erythropoietin levels and microcytosis in heterozygous beta-thalassaemia. AB - Erythropoietin levels were determined in 50 Greek females: 20 beta-thalassaemia (beta-thal) heterozygotes, 15 with a diagnosis of iron-deficiency anaemia and 15 normal controls. In beta-thal trait carriers, the erythropoietin levels were slightly higher than in normal controls (16.65 +/- 4.43 vs. 12.84 +/- 2.47 mU/ml); these levels were significantly lower than those in iron-deficient subjects with the same degree of anaemia (55.24 +/- 31.35 mU/ml). In both groups, the erythropoietin levels are statistically correlated with the severity of anaemia (r = -0.537 p < 0.05 for iron deficiency; r = -0.610 p < 0.01 for beta thal heterozygotes). In beta-thal heterozygotes, a close inverse correlation with red cell number and erythropoietin levels was also noted. It is suggested that microcytosis accompanying beta-thal trait constitutes an additional factor intervening in the regulation of erythropoiesis. PMID- 9352747 TI - Lymphoid blast crisis during complete cytogenetic remission following interferon alpha and hydroxyurea therapy. AB - Chronic myelogenous leukemia (CML) is a clonal disorder starting with a chronic phase and progressing to an acute blastic phase. Philadelphia (Ph) chromosome formation results in the relocation of the ABL oncogene from the chromosome 9q34 to BCR region on 22q11, forming the BCR/ABL fusion gene. The Ph chromosome once detected rarely disappears, except as a result of therapy. We present an unusual Ph-positive CML case, which developed lymphoid blast crisis in complete cytogenetic remission following interferon-alpha and hydroxyurea therapy. Sequential cytogenetic investigations were carried out on bone marrow. After a standard Ph translocation seen at diagnosis, from the 8th month of therapy all metaphases showed a normal diploid karyotype. Fluorescence in situ hybridization detected residual BCR/ABL-positive interphase cells during the 12th month of therapy. In the 14th month, the patient showed 27% blasts in marrow though normal cytogenetics was maintained. Present findings suggest blastic transformation occurred in a Ph-negative lymphoid clone. This supports the hypothesis that an actual leukemogenic event occurs in a multipotent stem cell prior to the acquisition of Ph translocation. PMID- 9352748 TI - Bone marrow hemophagocytosis and immunological abnormalities in a patient with lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is an inborn error of amino acid transport characterized by a wide spectrum of clinical and biochemical abnormalities. Bone marrow hemophagocytosis in this disorder is an intriguing finding, present mostly in Italian patients. We report a 19-month-old Turkish infant with LPI, bone marrow hemophagocytosis, interstitial lung disease and immunological abnormalities unprecedented in the current literature. Possible etiologic factors responsible for hemophagocytosis and the differential diagnosis of hemophagic syndromes are discussed. PMID- 9352746 TI - Natural killer cell frequency and serum cytokine levels in monoclonal gammopathies: correlation of bone marrow granular lymphocytes to prognosis. AB - The percent of granular lymphocytes of total bone marrow lymphocytes was 12.5% in controls, 15% in myeloma and 27% in monoclonal gammopathy of undetermined significance (MGUS). A good correlation was found between the percent of granular lymphocytes in the bone marrow lymphocytes at diagnosis (Y) and the years of survival (X) from the diagnosis of either the IgG- or IgA-type myeloma. The linear regression equation calculated for the IgG-type myeloma was Y = 1.6X + 7.42, and for the IgA-type myeloma Y = 4.25X + 4.75. For the purpose of analyzing in detail the granular lymphocyte behavior, two-color analyses of peripheral blood mononuclear cells and the serum levels of cytokines were performed. The absolute number of CD3+ cells, CD4+CD45RA+ cells and CD4+CD29+ cells was lower in the multiple myeloma (MM) cases than that of MGUS or controls (p < 0.01). The CD57+CD16+ natural killer (NK) cells were lower in MM cases than in MGUS cases. The serum levels of IL-1, which may activate NK cells, were higher in the MGUS cases than in either myeloma cases or controls (p < 0.01). The IL-10 levels, which may inhibit the proliferation of NK cells, were higher in the myeloma cases than in the MGUS cases (p < 0.05). Detailed understanding of the cytokine network of myeloma patients and their NK cell frequency may be important for the investigation of M proteinemias and for the future strategic planning of biological modulation therapies of myeloma patients. PMID- 9352749 TI - Chronic neutrophilic leukemia evolving from a myelodysplastic syndrome. AB - Chronic neutrophilic leukemia (CNL) is a rare hematologic disorder usually presenting with a persistent neutrophilia in the leukemoid range (WBC > 40-50 x 10(9)/1) and consisting largely of mature neutrophils. Patients have no obvious cause for an elevated white count and typically have an elevated leukocyte alkaline phosphatase score, hepatosplenomegaly, elevated vitamin B12 and are Philadelphia chromosome-negative. CNL has occasionally been associated with paraproteinemia or outright myeloma. Dysplastic features within the neutrophils in CNL have rarely been reported. We report the clinical, pathological and cytogenetic features of a case of CNL in an elderly white female initially diagnosed with refractory anemia with excess blasts, which subsequently progressed to CNL. PMID- 9352750 TI - Invasive pulmonary aspergillosis complicated by subclavian artery occlusion following allogeneic stem cell transplantation. AB - We describe an unusual case of invasive pulmonary aspergillosis (IPA) complicated by subclavian artery occlusion in a 32-year-old man with severe aplastic anemia, who underwent allogeneic stem cell transplantation. He was severely neutropenic after the conditioning for transplantation, but he had no history of fungal infection. Five days after the transplantation, he developed IPA in the left upper lung, complicated by left subclavian artery occlusion. Extensive chronic graft-versus-host disease, which required the administration of potent immunosuppressants for a long period of time, interfered with the resolution of the IPA. PMID- 9352751 TI - Hemoglobin sickle-lepore: an unusual case of sickle cell disease. PMID- 9352752 TI - Vascular thrombotic problems in Behcet's disease. PMID- 9352753 TI - Haemophagocytosis in bone marrow aspirates. PMID- 9352754 TI - Clearing the airway--the development of the pharyngeal airway. PMID- 9352755 TI - Continuous measurement of arterial and end-tidal carbon dioxide during cardiac surgery: Pa-ETCO2 gradient. AB - There have been reports of a negative arterial to end-tidal CO2 gradient (Pa ETCO2) during cardiac surgery, so we used capnometry and an intravascular blood gas sensor (Paratrend 7) to continuously monitor this gradient in 20 cardiac surgical patients. We also compared the values obtained from this sensor with those obtained from a standard blood gas analyser at seven time points. We found a significant change in Pa-ETCO2 after cardiopulmonary bypass (P < 0.001) though we were unable to demonstrate a negative Pa-ETCO2 at any time (95% CI 0-14%). There was clinically acceptable agreement between laboratory and Paratrend 7 measurements during and after cardiac surgery. PMID- 9352756 TI - Respiratory drive and pulmonary mechanics during haemodialysis with ultrafiltration in ventilated patients. AB - The improvements of respiratory drive and pulmonary mechanics which follow haemodialysis with ultrafiltration in mechanically ventilated renal failure patients seem predictable but have not been studied before. In this study, 14 renal failure patients with stable haemodynamics mechanically ventilated with pressure support ventilation (PSV) were enrolled. Respiratory drive (represented as P0.1), pulmonary mechanics, breathing pattern, arterial blood gas and haemodynamics were measured according to the time schedule: pre-dialysis (Time 0), and at 60, 120, 180, 240 minutes thereafter. Following the removal of excess lung water during haemodialysis, auto-PEEP and patient's work of breathing (WOBp) decreased gradually. P0.1 lessened progressively along with the improvement in pulmonary mechanics. The changes in auto-PEEP and WOBp correlated closely to the pre- and post-dialysis decline of P0.1 (delta P0.1). There was a negative, moderately significant correlation between the amount of fluid ultrafiltrated during dialysis (delta UF) and the delta P0.1 (R = -0.54). The breathing pattern remained stable during dialysis. No hypoventilation or hypoxaemia occurred despite the development of metabolic alkalosis induced by bicarbonate dialysate. We have shown that respiratory drive decreases gradually during bicarbonate haemodialysis. The improvements of pulmonary mechanics, rather than the rapid alkalization of body fluids, responds to the decrease of P0.1 in renal failure patients ventilated with PSV. PMID- 9352757 TI - Dynamic hyperinflation: comparison of jet ventilation versus conventional ventilation in patients with severe end-stage obstructive lung disease. AB - Positive pressure ventilation in patients with obstructive lung disease may result in over-inflation of the relatively compliant lungs, resulting in dynamic hyperinflation (DHI). Using a crossover trial design, we compared high-frequency jet ventilation (HFJV) versus "optimal" intermittent positive pressure ventilation (IPPV) in ten patients undergoing lung transplantation for severe, end-stage obstructive lung disease. We measured haemodynamics and the degree of DHI after both modes of ventilation. There were no significant differences between IPPV and HFJV, with respect to efficiency of ventilation (PaCO2), haemodynamic effects (stroke volume, blood pressure and cardiac output), or lung hyperinflation (trapped gas volume). This study suggests that HFJV, when compared with optimal IPPV, is no better at minimizing DHI in patients with severe, end stage obstructive lung disease. PMID- 9352759 TI - Predicting difficult intubation--a comprehensive scoring system. AB - A study was conducted in an attempt to devise a simple and more accurate method of predicting difficult intubation. Prospective assessments were made in 282 patients and retrospective assessment in 16 patients with regard to 21 anatomical factors which were correlated with the laryngoscopic view at intubation. Twelve factors correlated significantly with difficult intubation. Four of these were eliminated after multifactorial analysis. A scoring system was devised, assigning points to each variable based on its discriminative value. A score of 6 or more correctly identified 22 out of the 23 difficult intubations and there were 50 false positives (sensitivity, specificity and PPV of 96%, 82% and 31% respectively). When negative scoring was done for factors favouring easy intubation, false positives were reduced to 36, but only 20 difficult cases could be identified correctly. PMID- 9352758 TI - Epidural infusion of bupivacaine 0.0625% plus fentanyl 3.3 micrograms/ml provides better postoperative analgesia than patient-controlled analgesia with intravenous morphine after gynaecological laparotomy. AB - One hundred and twenty women undergoing gynaecological abdominal operations were randomized to receive either epidural bupivacaine 0.0625% + fentanyl 3.3 micrograms/ml infusion (Group EPI, n = 57), or patient-controlled intravenous morphine analgesia (Group PCA, n = 54) for postoperative pain relief. The groups were comparable in demographic data, types and duration of operation. Group EPI achieved significantly lower verbal rating scale of pain (VRS) at rest at 0, 4, 12, 16, 20, 28 and 40th postoperative hours. The VRS during cough were also significantly lower in Group EPI at 0, 4, 8, 12, 28 and 36th postoperative hours. None of the patients had respiratory depression or hypotension. Nausea/vomiting occurred in 52.6%/33.3% of patients in Group EPI and 52.7%/37.0% in Group PCA. Most patients (84.2% in Group EPI and 72.2% in Group PCA) rated their pain management as "good". We conclude that epidural infusion of bupivacaine 0.0625% and fentanyl 3.3 micrograms/ml provide better analgesia than patient-controlled intravenous morphine after gynaecological laparotomy. PMID- 9352760 TI - Ventilator-CPAP with the Siemens Servo 900C compared with continuous flow-CPAP in intubated patients: effect on work of breathing. AB - The effects of continuous positive airway pressure (CPAP) provided by the Siemens Servo 900C ventilator were compared with a continuous flow system (CF-CPAP) in patients weaning from the ventilator. Thirteen patients were studied using both systems at a CPAP level of 0.5 kPa. Additional work of breathing (Wapp) and derived variables were determined in relation to the minute volumes of the patients. The Wapp imposed by the ventilator exceeded the Wapp of CF-CPAP in all patients. The difference in Wapp between ventilator- and CF-CPAP was greater at higher ventilatory needs. The increments in Wapp imposed by the ventilator were positively correlated with the actual end-expiratory pressures (EEP). The EEP increasingly exceeded the preset CPAP level of the ventilator at higher minute volumes. An inspiratory threshold due to a gradient between EEP and preset CPAP greatly increased the Wapp imposed by the ventilator. As this threshold was attributed to the resistance of the PEEP device of the ventilator, it indicates that the additional work related to the expiratory value should be taken into account when the Siemens Servo 900C ventilator is used for weaning purposes. PMID- 9352761 TI - Laser prostatectomy versus transurethral resection of the prostate for benign prostatic hypertrophy: comparative changes in haemoglobin and serum sodium. AB - A prospective study was undertaken to examine differences in haemoglobin concentration and serum electrolytes in two patient groups undergoing surgical treatment for benign prostatic obstruction. Group one underwent conventional transurethral resection of the prostate (TURP), and group two were treated by laser ablation of the prostate (LAP). Twenty-six patients were enrolled in LAP group, 25 in the TURP group. Both patient groups had the procedure performed under epidural anaesthesia. Serial measurements of haemoglobin and sodium were performed at three time intervals: immediately preoperatively, in the recovery room and 24 hours postoperatively. A fall in serum sodium levels between the mean preoperative reading (140 mmol/l) and 24 hours post surgery (138 mmol/l) was the only statistically significant alteration sustained in the laser patients (P < 0.0001). A fall in haemoglobin from preoperative measurement to recovery room measurement of 0.71 g/l was statistically significant (P < 0.0001), but did not persist to the 24 hour postoperative time period. The TURP group demonstrated statistically significant falls in both sodium and haemoglobin levels at both postsurgery measurements. Mean serum sodium levels fell from 141 mmol/l preoperatively to 138 mmol/l (P < 0.0001) in the recovery room and 137 mmol/l (P < 0.0001) at 24 hours. Preoperative haemoglobin fell from 14.8 g/l to 13.6 g/l (P < 0.0001) in recovery and 13.7 g/l (P < 0.0001) at 24 hours. PMID- 9352762 TI - Aspects of theophylline clearance in children. AB - Michaelis-Menten pharmacokinetic parameters for theophylline were estimated in a three-month infant following an accidental overdose of intravenous aminophylline. Fitting of time-concentration data was performed using nonlinear regression with MKMODEL. A mixed order elimination model was superior to a first order model. Parameter estimates were standardized to a 70 kg human using an allometric power model. Parameter estimates (SE) were: maximum rate of metabolism (Vmax) 71 (42) mg.h-1, Michaelis-Menten constant (Km) 32.3 (33.5) mg.l-1, volume of distribution (Vd) 46.9 (2.6)l. This Michaelis-Menten constant is lower than that reported for adults and consequently non-linear elimination will occur at lower plasma concentrations in infants than in adults. Theophylline clearance has traditionally been reported as directly proportional to body weight. This per kilogram model gives an erroneous impression that clearance is greatest in early childhood and then decreases with age until adult rates are reached in late adolescence. Age-related clearance values reported in the literature were reviewed using an allometric 3/4 power model. This size model demonstrates that clearance increases in infancy and reaches adult rates in the first one to two years of life. PMID- 9352763 TI - Acute weakness syndromes in critically ill patients--a reappraisal. AB - Over the last twenty years, increasing numbers of critically ill, mechanically ventilated patients who develop acute profound muscle weakness have been described. These acute weakness syndromes have not been well understood and they have been given many names including: acute steroid myopathy, acute quadriplegic myopathy, the floppy person syndrome, critical illness polyneuropathy, critical illness polyneuromyopathy, and prolonged neurogenic weakness. Many of these "syndromes" either overlap or represent the same disease process in different patients. Many have been incompletely diagnosed. During this review it became evident that the acute weakness syndromes currently recognized in critically ill patients could be categorized into four major groups: myopathy, neuromuscular junction abnormalities, neuropathy and polyneuromyopathy. Each had different possible aetiologies. "Myopathy" includes acute necrotizing myopathy and disuse atrophy. Neuromuscular junction abnormalities are subdivided into myasthenia-like syndromes and prolonged neuromuscular blockade. Neuropathies are divided into critical illness polyneuropathy and acute motor neuropathy. The anterior horn cell injury in Hopkins syndrome should also be considered in this group. Polyneuromyopathies include various combinations of neuropathy and myopathy in the same patients. PMID- 9352764 TI - Anaesthetists' attitudes towards an anaesthesia simulator. A comparative survey: U.S.A. and Australia. AB - Anaesthesia simulation has been suggested as a method to enhance the training of clinicians without exposing patient to risk. Recently, two anaesthesia simulators have become commercially available in the U.S.A. Attitudes towards anaesthesia simulators have not been previously surveyed. With institutional approval, a survey questionnaire was given to 1. all clinical staff of the Department of Anesthesiology, University of Pittsburgh Medical Center, and 2. all anaesthetists attending the Annual General Meeting of the Australian Society of Anaesthetists. An information sheet containing details about anaesthesia simulation in general and the special capabilities of a particular commercial anaesthesia simulator was included with the survey instrument. The survey was anonymous and contained 15 questions. Attitudinal responses were recorded using an anchored visual analog 100 mm scale. We surveyed anaesthetists during September-October 1993. Completed forms were returned by 183 anaesthetists. Respondents were aged 25-67 years (mean age 41 +/- 10 yr) and were grouped by staff position (78% faculty, 22% trainees), sex (79% male; 21% female), country of practice (44% Aust, 56% U.S.A.) and years in practice. Seventy-three per cent staff were in favour (VAS > 60) of departmental purchase of a simulator (with no significant difference between countries) and 76% expressed willingness (VAS > 60) to undergo testing in their own time (with Australian anaesthetists significantly more willing to do so). However, 65% were not in favour (VAS < 40) of the compulsory use of a simulator for re-certification or re-accreditation of anaesthesia practitioners, with American anaesthetists (anesthesiologists) significantly more opposed to it. The most frequent comment related to the cost. There is majority support for the purchase of an anaesthesia simulator but there is widespread concern for its high cost. In general, anaesthesia simulation is perceived more as an education tool rather than an instrument for (re)certification. PMID- 9352765 TI - Analgesia following thoracotomy: a survey of Australian practice. AB - This survey examines pain management after thoracotomy in Australian hospitals. Questionnaires were sent to senior thoracic anaesthetists at 27 hospitals (16 public and 11 private) with thoracic surgical units. Twenty-six anaesthetists replied and 24 responses were included in the analyses. Seventy-two percent of respondents were from hospitals with acute pain services (APS), and in 94% of these hospitals patients are reportedly visited by the APS. The most frequently used analgesic modalities are epidural analgesia, intravenous patient-controlled analgesia (IVPCA), and nurse-controlled intravenous opioid infusions. Over half of the anaesthetists reported using local anaesthetic intercostal nerve block, non-steroidal anti-inflammatory drugs (NSAIDs), or paracetamol. Combinations of analgesic techniques were cited frequently. Respondents reported that cryoanalgesia, interpleural blockade, paravertebral blockade, subarachnoid infusions, ketamine, and transcutaneous electrical nerve stimulation are used infrequently. Anaesthetists from public hospitals reported using epidural analgesia, IVPCA and NSAIDs more frequently than those from private hospitals. When epidural analgesia is used, most respondents place the catheter in the mid thoracic region (91%), use a regimen of opioids plus local anaesthetic (96%), use a constant infusion technique (100%), and continue analgesia for up to three days (83%). Over half of the respondents reported that post-thoracotomy patients are nursed in a high-dependency area. Seventy-nine percent of respondents selected epidural analgesia as the best available analgesia technique, whereas 21% consider IVPCA to be the best. Only 75% of respondents reported that the type of analgesia they consider best is also the type which they use most frequently. PMID- 9352766 TI - Relationship of muscle strength to potassium concentration in a hypokalaemic infant. AB - A nine-month infant, weighing 9.8 kg, presented with hypotonia secondary to acute hypokalaemia (1.0 mmol/l). Muscle strength improved as the serum potassium was increased. Muscle strength was assessed by the pressure generated inside a saline filled endotracheal tube cuff during a grasp reflex. Potassium concentration and hand grip strength were related using a sigmoidal Emax model. Zero effect was assumed when the potassium concentration was zero. The Emax, EC50 and Hill coefficient values were determined by non-linear regression using the MKMODEL program. Parameter estimates (SE) were EC50 1.79 (0.15) mmol/l, Hill coefficient 3.79 (0.92), Emax 114.4 (8.9) mmHg. PMID- 9352767 TI - Assumptions and practice in clinical medical ethics. AB - An orderly scheme of action is proposed to allow for the practical solution of clinical ethical problems. This scheme depends on understanding and discussion, between patient and doctor, of the ethical assumptions involved in any dilemma. Instead of the more usual ethical principles, arguments are presented for six basic ethical assumptions (and their associated corollaries) in favour of Life, Autonomy, Beneficence, Equity, Truth, and Law. Because these assumptions are dependent on the different personal viewpoints of the people involved and not immutable principles, such ethical assumptions are able to be set in different hierachical orders on different occasions permitting in most cases a particular solution specific for that dilemma. PMID- 9352768 TI - Perioperative management of intra-partum seizure. AB - A young nulliparous woman previously diagnosed with pregnancy-induced hypertension suffered a seizure during active first stage labour. Pre-seizure blood pressure was borderline high but she did not fulfill other criteria for preeclampsia. She underwent emergency caesarean section for presumed eclampsia. Postoperatively, she deteriorated neurologically. CT scan showed an intracranial haemorrhage requiring neurosurgical intervention. The similarities of presentation of a primary cerebrovascular event and eclampsia following an intra partum seizure made the differential diagnosis difficult. PMID- 9352769 TI - Tension pneumopericardium following tracheoplasty for congenital tracheal stenosis. AB - Tension pneumopericardium is a rare complication of mechanical ventilation following tracheoplasty for congenital tracheal stenosis. This case report describes fatal tension pneumopericardium in a three-month-old male infant some 24 hours following tracheoplasty for this condition. Because of persisting stenosis of the left main bronchus, ventilation pressures of PIP 23 cm H2O and PEEP 5 cm, progressing to PIP 28 cm and PEEP 7 cm were needed to maintain adequate respiratory exchange. Partial relief of the pneumopericardium was achieved by vigorous cardiac massage, but three days later the infant died of massive airway haemorrhage. PMID- 9352771 TI - Intraoperative convulsions in a child with arthrogryposis. AB - A case of intraoperative convulsions occurring in a child with arthrogryposis multiplex congenita is presented. Arthrogryposis and the anaesthetic management of children with this condition is discussed. Factors which may have contributed to the convulsions are considered. PMID- 9352770 TI - Tricyclic poisoning--successful management of ventricular fibrillation following massive overdose of imipramine. AB - Serious complications from tricyclic antidepressant (TCA) overdose are uncommon. We present a case of massive imipramine overdose complicated by ventricular fibrillation and a prolonged period of cardiovascular collapse. A total of 400 mmol of sodium bicarbonate, 5 mg of adrenaline and 80 mg of sotalol were given during 50 minutes of cardiac arrest. The patient made a full recovery with no apparent neurological sequelae. The highest TCA plasma level we could find in the published literature was 4873 ng/ml4; our patient's peak TCA level was 6000 ng/ml. Tricyclic antidepressant overdose is a common cause of intensive care unit admission. It has a low mortality rate. PMID- 9352772 TI - Unilateral pulmonary oedema following laryngospasm. PMID- 9352773 TI - Subarachnoid haemorrhage presenting as postoperative neck pain. PMID- 9352774 TI - Avoiding pulmonary artery rupture. PMID- 9352775 TI - Acupuncture and postoperative vomiting in day-stay paediatric patients. PMID- 9352776 TI - Propofol infusion for the difficult airway. PMID- 9352777 TI - Fatal outcome after propofol sedation in children. PMID- 9352778 TI - Propofol and malignant hyperthermia susceptibility. PMID- 9352779 TI - Fatal outcome after propofol sedation in children. PMID- 9352780 TI - Needle-through-needle technique. PMID- 9352781 TI - Hypothermia associated with subarachnoid morphine. PMID- 9352783 TI - Oximetry and patent blue five dye. PMID- 9352782 TI - Ecstasy: creatinine kinase. PMID- 9352785 TI - Gauze bite block. PMID- 9352784 TI - Hyperkalaemic cardiac arrest following succinylcholine in a longterm intensive care patient. PMID- 9352786 TI - Defibrillator pad for airway management in epidermolysis bullosa--saving face. PMID- 9352787 TI - Graseby 3300 PCA pumps. PMID- 9352789 TI - Acute myeloid leukemia following psoralen with ultraviolet A therapy: a fluorescence in situ hybridization study. AB - A woman with mycosis fungoides treated by psoralen with ultraviolet A (PUVA) and electron beam therapy developed acute myeloid leukemia (AML) three years later. Karyotypic analysis of the leukemia cells revealed monosomy 7. Fluorescence in situ hybridization showed that the monosomy 7 clone had accounted for about a third of the marrow cells after PUVA treatment, but replaced the entire marrow at leukemic transformation. These findings were consistent with a secondary AML evolving from an underlying myelodysplasia, supporting that PUVA therapy might have a mutagenic effect on hematopoietic cells. This might be related to its effect on circulating hematopoietic stem cells. PMID- 9352788 TI - Establishment and characterization of a highly tumorigenic human diploid endometrial cancer cell line. AB - A new cell line designated CUME-1 has been established from a poorly differentiated endometrial adenocarcinoma of the uterus. This cell line grew well without interruption for more than 88 months and 110 serial passages were successively carried out. The cells were highly tumorigenic in nude mice (85%). Repeated karyotype analyses from early (4th) to late (55th) passages of this cell line revealed a diploid stable clone in each passages without any noticeable structural or numerical aberrations. But from the 80th passage, a subpopulation with reciprocal translocation between chromosomes 1q and 9q consistently appeared and was observed in about 30% of the cells. This cell line is one of the rare examples of experimentally proved tumorigenic cells of human solid tumor origin that retains the diploid karyotype in vitro. HLA typing indicated the presence of DR4, DR13, DQ3 and DQ6. Cytosol estrogen and progesterone receptors were found both in fresh primary tumor and in this cell line. Gonadotropin-releasing hormone (Gn-RH) receptor mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) in cultured cells. Using the single-strand conformation polymorphism (SSCP) technique, we have screened CUME-1 cells for p53 mutation in exons 4 to 9. No mobility shift was observed. This cell line may be useful in studying the in vitro chromosomal evolution of the cell line and the in vivo properties of human endometrial adenocarcinoma. PMID- 9352790 TI - Overrepresentation of 1q21-23 and 12q13-21 in lipoma-like liposarcomas but not in benign lipomas: a comparative genomic hybridization study. AB - Twenty lipomatous tumors, including eight lipoma-like liposarcomas and 12 benign lipomas, were analyzed using comparative genomic hybridization (CGH). DNA sequence copy number changes detected in five lipoma-like liposarcomas (mean, 1.1 aberrations/tumor; range, 0-2) consisted of gains of 12q13-21 (five tumors) and 1q21-23 (four tumors). Two of the tumors showed high-level amplification at 12q14 21 and one tumor at 1q21-22. No copy number changes were found in lipomas. Overrepresentation of 1q and 12q sequences was a recurrent finding in lipoma-like liposarcomas but not in lipomas. Thus, CGH may help in the differential diagnosis of low-grade or borderline adipose neoplasms. PMID- 9352791 TI - Acute myeloid leukemia with trisomy 11: a molecular cytogenetic study. AB - Trisomy 11 is uncommon in acute myeloid leukemia (AML) and molecular studies have shown partial tandem duplication of the MLL gene in some cases. In a case of AML with trisomy 11, the MLL gene was found to be tandemly duplicated, leading to the formation of a fusion transcript involving splicing of exon 8 to exon 2. Fluorescence in situ hybridization revealed two populations of blasts, with about two-thirds of them showing trisomy 11. These findings suggested that the trisomic and non-trisomic clones had evolved from a clone with a common submicroscopic mutation. As recent studies showed that MLL gene duplication in fact occurs more often as a primary mutation in the absence of trisomy 11, it is possible that in our case the MLL gene duplication might be the common underlying mutation. The clinical course of this case was similar to the poor prognosis reported for trisomy 11. PMID- 9352792 TI - A subset of gestational trophoblastic disease characterized by abnormal chromosome 8 copy number detected by fluorescence in situ hybridization. AB - The present paper describes the results of research conducted to ascertain whether the report by Mark et al. [1], describing the concurrence of congenital trisomy 8 mosaicism and gestational trophoblastic disease (GTD) in a 42 year-old Gravida IV, Para IV patient was an isolated event. In contrast to other cases described in the literature, the patient described in Mark et al. [1] had no additional confounding chromosomal abnormalities other than trisomy 8. To the best of our knowledge, ours was the only reported case of constitutional trisomy 8 mosaicism associated with gestational trophoblastic disease, a rare gynecological disease entity. The question arises whether there exists a subset of patients with GTD characterized by an abnormal chromosome 8 copy number. The implicit hypothesis is that an abnormal number of chromosome 8 somehow predisposes to cancer. A pilot study of 10 cases of GTD was conducted using fluorescence in situ hybridization (FISH) and a commercial chromosome 8-specific alpha-satellite probe on formalin-fixed, paraffin-embedded patient tissues. Among eight informative cases successfully completed, two cases (25%) were found to be trisomic, when a cut-off point of 10% trisomic cells is adopted. Another two cases (25%) were found to be triploid. The results of our FISH study indicated that an abnormal chromosome 8 copy number found in Mark et al. [1] is unlikely to be an isolated event. Our data are consistent with the hypothesis that a subset of GTD indeed may exist which is characterized by more than two copies of chromosome 8. The present findings corroborate those recently found in breast, prostate, and other cancers. PMID- 9352793 TI - 12p rearrangement and DNA amplification mapped by comparative genomic hybridization in a patient with secondary myeloid leukemia. AB - Rearrangements of the short arm of chromosome 12 (12p) are a common finding in hematologic malignancies. There has recently been considerable interest in chromosome 12 abnormalities in view of the mapping of the TEL gene to 12p13 and frequent 12p interstitial deletions. Overrepresentation of 12p sequences is, on the other hand, a consistent finding in testicular germ cell tumor (TGCT), and the 12p11.2-p12.1 subregion has been found to be specifically involved. We have studied a secondary leukemic patient whose cells contained 12p rearrangements with a view to clarifying the underlying molecular events. Fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH) have revealed the presence of 12p11 breakpoints on both 12 homologs as well as amplification of 12p11-p12-derived sequences. Six YACs and a cosmid probe have been used in an attempt to map the amplification unit on 12p. The two YACs contigs WC-1468 and WC 985 were not amplified, and our results suggested a small amplicon localized in the 12p11.2-p12 subregion. We speculate that this region harbors gene(s) which are critical in tumor formation and could be involved in both TGCT and our patient. Whether the same gene(s) are involved in both amplification and translocation is unknown. PMID- 9352794 TI - Chromosome aberrations in renal tumors detected by fluorescence in situ hybridization. AB - The aim of this study was to investigate the relationship between chromosome aberrations detected by fluorescence in situ hybridization (FISH) and tumor grade, stage, venous involvement, and DNA ploidy status in 18 renal tumors. Using FISH with chromosome-specific DNA probes, the copy number of pericentromeric sequences on chromosomes 3, 7, 9, and 17 was detected within interphase nuclei in touch preparations from tumor specimens. Monosomy for chromosome 3 was detected in seven of 9 DNA diploid tumors, whereas all DNA aneuploid tumors demonstrated trisomy or tetrasomy for chromosome 7. Moreover, monosomy for chromosome 3 was more frequently shown in the diploid and low-stage tumors than in the aneuploid and high-stage tumors. The percentage of hyperdiploid cells significantly correlated with DNA ploidy status in the case of chromosomes 3 and 7 (p = 0.030, p = 0.007, respectively). The percentage of hyperdiploid cells for chromosome 3 had borderline significance with tumor stage. On the other hand, the percentage of diploid cells for chromosome 17 was significantly correlated with DNA ploidy status and tumor stage (p = 0.030, p = 0.027, respectively). Moreover, the percentage of diploid cells for chromosome 7 in renal cell carcinoma (RCC) with venous involvement was significantly lower than those without venous involvement (p = 0.023). These results suggest that the incidence of chromosomal aberrations detected by FISH is more frequent than the chromosomal aneuploidy reported previously by conventional cytogenetics. Therefore, loss of chromosome 3 may be associated with an early event in RCC carcinogenesis. Gain of chromosomes 3 and 7 is correlated with tumor progression as well as gain and loss of chromosome 17. Study of the chromosomal aberrations may provide a greater understanding of tumor carcinogenesis and progression in RCC. PMID- 9352795 TI - Cytogenetic aberrations and DNA ploidy in soft tissue sarcoma. A Southwest Oncology Group Study. AB - We performed cytogenetic analysis and determined DNA content by flow cytometry (FCM) on freshly disaggregated tumor biopsies from 45 patients with soft tissue sarcomas (STS). Cytogenetically aberrant clones characterized 30 (67%) tumors, with the remaining 15 yielding normal karyotypes with or without nonclonal aberrations. No tumors with multiple unrelated clones were observed. Among the 30 tumors with clonally abnormal karyotypes, 21 (70%) had near-diploid stemlines, six were near-triploid and three were near-tetraploid. Ten of the clonally aberrant tumors contained nonrandom chromosomal translocations characteristic of histologic subtypes. Overrepresentation of chromosomes 7 and 8 were common numerical aberrations. Structural aberrations most often involved chromosomes 1, 7, 9, 12, and 14. Clustering of breaks in 9p resulting in partial loss of the short arm was frequent. Unstable aberrations including rings, dicentrics, large markers, small numbers of double minutes, and telomeric associations were seen in nine tumors. With FCM, 27 (60%) tumors had aneuploid DNA content and 18 (40%) were DNA diploid. Of those 18 DNA diploid tumors, 11 showed clonal karyotypic aberrations. In addition, apparent discrepancies between the results of the cytogenetics and FCM with respect to ploidy pattern were seen in 13 samples; 11 had DNA content in the peritriploid to peritetraploid range but the corresponding karyotype was normal or near-diploid. When the findings of the cytogenetics and DNA content analyses were combined, an abnormal cell population by one or both methods was detected in 38 (84%) tumors. The concurrent application of standard cytogenetics and DNA ploidy by FCM provide complementary information confirming a high incidence of genetic alterations in STS. PMID- 9352796 TI - Cytogenetic study of twenty-three primary squamous cell carcinomas of the lung. AB - Fifty-seven primary squamous cell carcinomas of the lung were analyzed cytogenetically. Karyotyping was possible in seven cases, and chromosome counting without detailed analysis was possible in 16 other cases. The results suggested that structural chromosome rearrangements related to the short arms of chromosomes 1(5/7), 9(3/7), and 11(6/7), and the long arms of chromosomes 6(4/7) and 7(6/7) may be the primary and non-random chromosome defects which are closely associated with human lung squamous cell carcinoma. These primary and non-random chromosome defects are believed to confer a proliferative advantage to cells carrying them, and to be involved in the pathogenesis of human lung squamous cell carcinoma. PMID- 9352798 TI - A second case of hexasomy 8 in myelodysplastic syndrome. AB - Tetrasomy 8 is a rare form of acquired aneuploidy found exclusively in the myeloid leukemias. Hexasomy 8 is even rarer: only one case has been reported, thus far. We describe here the second case of hexasomy 8 as the sole abnormality in an elderly female patient with myelodysplastic syndrome (MDS). PMID- 9352797 TI - Comparison of the chromosomal pattern of primary testicular nonseminomas and residual mature teratomas after chemotherapy. AB - About 70 to 75% of patients with nonseminomatous testicular germ cell tumors (NSs) present with metastases. When these metastases are treated with chemotherapy, often residual mature teratoma (RMT) is left. RMT is composed of fully differentiated somatic tissue. Untreated metastases of NSs rarely consist exclusively of mature somatic tissue. Apparently, after chemotherapy treatment there is a shift towards higher degrees of differentiation. Investigating tumor progression and the mechanism(s) involved in therapy-related differentiation, we compared the cytogenetically abnormal karyotypes of a series of 70 NSs with those of 31 RMTs. In NSs and RMTs, the modal total chromosome number does not differ and is in the triploid range. Both the frequency and the average copy number of i(12p) are the same, and the pattern of chromosomal over- and underrepresentation and distribution of breakpoints do not differ significantly in these series. So, we found the chromosomal pattern of RMTs as abnormal as those of primary NSs. Based on cytogenetics, we found no indication that specific chromosomal alterations parallel metastasis and therapy-related differentiation of the metastases. The cytogenetic data suggest that both induction of differentiation of (selected) cells or selection of cells with capacity to differentiate are possible mechanisms for the therapy-related differentiation of RMTs. PMID- 9352799 TI - Cytogenetic and interphase cytogenetic analyses reveal chromosome instability but no clonal trisomy 8 in Dupuytren contracture. AB - The results of cytogenetic and FISH analysis performed in 26 cases of Dupuytren contracture are reported. Clonal or sporadic chromosome changes were found in 18 cases (69%). Clonal changes consisted of: +2, +16, -10, -Y, add(1)(p23), del(2)(q21), t(3;16)(p21;q24), add (3)(p24), del(18)(q21), t(Y;14)(p12;q24), +mar. The results differ from those obtained in normal palmar fascia used as control, in which -Y and +Y were the only clonal changes found in 2 of 11 analyzed cases (18%). No clonal trisomy 8 was found. FISH analysis performed in 11 cases (centromeric probe specific for chromosome 8) failed to show the presence of a cell population with +8. Clonal and sporadic structural changes were different from case to case and no clustering breakpoint was observed. The significance of the chromosome instability leading to clonal and sporadic chromosome changes not specific to Dupuytren contracture are discussed. PMID- 9352800 TI - Ph-positive CML in blastic phase with monosomy 7 in a Down syndrome patient. Monitoring by interphase cytogenetics and demonstration of maternal allelic loss. AB - We report a case of Ph-positive chronic myelocytic leukemia in blastic phase in an 11-year-old boy with Down syndrome. Monosomy 7 was the only additional chromosomal anomaly in the blastic clone. Fluorescence in situ hybridization analysis on interphase nuclei with a centromeric probe specific to chromosome 7 proved to be efficient in disease monitoring, and showed, together with the results of chromosome analysis on metaphases, that B-lymphocytes at the origin of an EBV-established line were not part of the leukemic clone. The study of DNA polymorphisms showed that the origin of the constitutional trisomy 21 was a maternal anaphase I nondisjunction, that the chromosome 7 lost in the blastic marrow clone was the maternal one, and led us to postulate that the mother's chromosomes are prone to impairment of normal disjunction. The study of allelic losses of chromosome 7 loci proved to be a further possibility for disease monitoring. PMID- 9352801 TI - Chromosome breakpoint distribution in nonmelanoma skin cancers. AB - We have identified chromosome regions that may be sites of genes activated as a result of chromosomal rearrangements observed in 61 of the 86 skin tumors referenced in the literature. The data showed that most of the breakpoints were distributed throughout the genome and some tended to cluster. Highest frequencies of breakpoints were observed in chromosomes with high relative length, except chromosomes 14 and 15 that were often affected in malignant tumors, despite their size. Our work provides a starting point for more detailed studies that may allow identification of these genes as important keys in the development and progression of skin cancers. PMID- 9352802 TI - Myelodysplastic syndrome with biclonal monosomy 7 and trisomy 8 after treatment with cladribine (2-chloro-2-deoxyadenosine) and involved field radiation therapy. AB - Therapy-related myelodysplastic syndrome (t-MDS) and acute nonlymphocytic leukemia (t-ANLL) are dramatic complications of cancer chemotherapy. Drugs like plant alkaloids or antimetabolites have not been reported to cause either t-MDS or t-ANLL. Monosomy 7(-7) and trisomy 8(+8) are among the most common abnormalities in myelodysplastic syndromes. Both abnormalities in two different clones of the same patient are very rarely reported. Such a myelodysplastic syndrome occurring shortly after treatment with an antimetabolite, the adenosine analogue cladribine (1-chlorodeoxadenosine), and involved field radiotherapy is reported here. PMID- 9352803 TI - Endoscopically assisted open removal of laterally herniated lumbar discs. AB - BACKGROUND: Adequate treatment of laterally herniated lumbar discs presents a surgical challenge. All fragments compressing the nerve root should be removed without destruction of the overlying facet joint. To accomplish this goal many techniques have been proposed, each with specific limitations. METHODS: In a small group of patients we have used a small malleable endoscope to assist in the removal of laterally herniated discs. Via a small laminotomy, the nucleus was removed in a standard manner and then the lateral disc material removed under direct endoscopic visualization. RESULTS: There were no operative complications and the nerve root was visualized and decompressed in all patients. There has been no recurrence of pain or development of spondylolisthesis. CONCLUSION: This technique negates removal of portions of the facet joint and provides adequate nerve root decompression. PMID- 9352805 TI - Vascularized pedicled laminoplasty. AB - BACKGROUND: Various advantages of a laminoplasty over the conventional laminectomy have been listed in the recent literature. An alternative method of vascularized pedicled laminoplasty is described in this report. METHODS: A linear cut is made in the laminae in their lateral aspects and two or more laminae are elevated as a flap and pedicled on supraspinous, interspinous, and interlaminar ligaments. The procedure was performed in 10 cases where surgery was performed for spinal intradural pathology. The ages of these patients ranged from 21 to 60 years. RESULTS: In an average observation period of 8 months, no demonstrable resorption of the laminar flap was observed in any case. None of the patients complained of any additional discomfort, pain, or deformity that could be related to the laminoplasty. CONCLUSIONS: Laminoplasty using laminae with a vascularized pedicle as described may be better tolerated by the patient; the segment seems to be more resistant to infection and radiotherapy. It is a simple, safe, and stable alternative to the procedures of laminoplasty previously described. No sophisticated instrumentation, prosthetic material, or technically complex maneuvers are required to harvest and reposition the laminar flap. PMID- 9352804 TI - Efficacy of surgical treatment in traumatic central cord syndrome. AB - BACKGROUND: Controversy surrounds the treatment of traumatic central cord syndrome (TCCS), as there are strong advocates for nonsurgical treatment for most patients. However, conservative treatment has been shown to yield a longer period of discomfort from pain and weakness in certain cases. METHODS: In a retrospective review of 114 patients presenting with acute or chronic TCCS from 1988-94, four different age groups were separately observed under different treatments. Motor and sensory recovery were assessed. RESULTS: Better results were achieved in younger patients, with or without radiographic abnormalities, and in patients with clinically correlated encroaching cord lesions who received early surgical decompression. CONCLUSIONS: Surgical intervention for TCCS must be addressed with careful clinical and radiographic survey. Removal of offending lesions in the subacute period results in significant motor and sensory improvement in short-term and long-term follow-up. PMID- 9352806 TI - Neurosurgery of the peripheral nervous system: injuries, degeneration, and regeneration of the peripheral nerves. PMID- 9352807 TI - Middle cerebral artery aneurysm complicated by tuberculosis. PMID- 9352808 TI - Intra-aneurysmal pressure changes during angiography in coil embolization. AB - BACKGROUND: Although elevation of blood pressure in aneurysms induced by injection of contrast medium has been postulated as a major cause of rerupture of ruptured cerebral aneurysms during angiography, no study has proved the elevation of intra-aneurysmal pressure because of difficulty in measuring the intra aneurysmal pressure during angiography. The present study demonstrated intra aneurysmal pressure to be raised by injection of contrast medium, using a microcatheter introduced into aneurysms. METHODS: To confirm the accuracy of pressure measurement through a microcatheter, we measured intra-aneurysmal pressure in a plastic model of an artery and an aneurysm during and after injection of contrast medium through a microcatheter and a needle inserted into the aneurysm. In a clinical study, intra-aneurysmal pressures were measured through the microcatheter in nine cerebral aneurysms of seven patients. RESULTS: In the model experiment, changes in the pressure measured through the microcatheter correlated well with those observed through the needle. In the clinical study, intra-aneurysmal systolic pressures increased by 5-23 mm Hg immediately after injection of contrast medium for 1-3 s in four basilar tip, three internal carotid-ophthalmic, and one middle cerebral artery aneurysm, whereas no pressure change was observed in a posterior cerebral artery aneurysm. Systemic blood pressure during angiography remained unchanged in all cases. CONCLUSIONS: This abruptly elevated intra-aneurysmal pressure by injection of contrast medium might cause rerupture of an aneurysm soon after rupture of the aneurysm, especially when the rupture site is fragile. PMID- 9352809 TI - Operation on high-lying basilar bifurcation aneurysms. AB - BACKGROUND: The surgical approach to basilar bifurcation aneurysms is a complicated procedure, especially with those located high in the interpeduncular cistern. We have developed a surgical approach to these aneurysms involving only a small anterior temporal craniotomy and detachment of the zygomatic arch. METHODS: The skin incision is placed 5 mm below the inferior border of the zygomatic arch and 5 mm anterior to the tragus, extending on towards the eyebrow but well within the hairline. The zygomatic arch is detached on either side followed by a small temporal craniotomy exposing the anterior temporal lobe. The temporal lobe is retracted and the internal carotid artery, posterior cerebral artery, posterior communicating artery, and inferior aspect of the optic chiasma are exposed. RESULTS: We have performed a transzygomatic anterior subtemporal approach in 12 patients with high-lying basilar bifurcation aneurysm. Eight patients demonstrated a good recovery as per the Glasgow outcome scale and four patients had a moderate disability that correlated with a poor WFNS grade preoperatively. One patient had an oculomotor palsy and another experienced a small infarction postoperatively. CONCLUSIONS: This approach is a suitable procedure for high-lying basilar bifurcation aneurysms in the interpeduncular cistern and 1.2 cm above the clinoid process. PMID- 9352810 TI - Asymptomatic, unruptured carotid-ophthalmic artery aneurysms: angiographical differentiation of each type, operative results, and indications. AB - BACKGROUND: Some types of carotid-ophthalmic artery aneurysms are still difficult to clip successfully because their exposure requires opening the cavernous sinus and/or retracting the optic nerve. It is useful to know the complications and to determine the type of aneurysm preoperatively for the management of carotid ophthalmic artery aneurysms. METHODS: The operative results in 15 patients with asymptomatic unruptured carotid-ophthalmic artery aneurysms were surveyed. The aneurysms were small in all the patients, and they underwent direct operation. Four patients presented with other ruptured aneurysms, four with other diseases (infarction, trauma, or pituitary adenoma), and seven were evaluated with magnetic resonance angiography for symptoms such as vertigo or headache. Among them, five had carotid cave aneurysms and one had paraclinoid aneurysm. RESULTS: Neck clipping was performed in 13 patients. Postoperatively, ipsilateral visual loss was encountered in one patient, and ipsilateral visual field defect was encountered in three patients. The visual field defect was lower nasal quadrant hemianopsia in two patients and lower hemianopsia in one patient. The cause of this complication was suspected to be retraction and/or the heat of the drill near the optic nerve. It seemed to be possible to distinguish the carotid cave or the paraclinoid aneurysm from the other carotid-ophthalmic aneurysms using carotid angiography preoperatively. CONCLUSION: When direct operation is performed for a carotid-ophthalmic artery aneurysm, care must be taken to avoid optic nerve injury caused by the retraction and/or the heat of the drill. PMID- 9352811 TI - Obliteration of a giant carotid aneurysm after extracranial-to-intracranial bypass surgery: case report. AB - BACKGROUND: Proximal arterial occlusion, with or without extracranial-to intracranial (EC-IC) bypass, is frequently used as treatment for giant intracranial aneurysms that are unclippable. The authors report on a patient who had obliteration of a giant unruptured aneurysm of the right internal carotid terminus after undergoing an EC-IC bypass without proximal arterial ligation. METHODS: This 71-year-old woman presented with repeated right cerebral ischemia caused by a giant saccular aneurysm of the right internal carotid terminus. Direct surgical clipping of the aneurysm was not recommended because of the patient's age and because of the morphology of the aneurysm. She could not tolerate occlusion of the right internal carotid artery (ICA) and, therefore, first underwent an EC-IC bypass. Four weeks later, she returned to undergo a balloon occlusion of the right ICA proximal to the aneurysm. RESULTS: The right distal ICA and aneurysm were found to be spontaneously thrombosed. At 2-year follow-up, the aneurysm was shown to be completely obliterated on the magnetic resonance imaging scans. CONCLUSIONS: The authors conclude that hemodynamic changes in the blood flow of the parent artery after EC-IC bypass caused this occurrence. PMID- 9352812 TI - Intracranial dissecting aneurysm causing subarachnoid hemorrhage: the role of computerized tomographic angiography and magnetic resonance angiography. AB - BACKGROUND: With increasing frequency, dissecting aneurysms of the intracranial arteries are recognized as a possible cause of subarachnoid hemorrhage (SAH). In the presence of a dissecting aneurysm, angiographic changes may be subtle at presentation and correct diagnosis often requires serial angiograms. We report a patient with a dissecting aneurysm of the anterior cerebral artery (ACA) causing SAH, in whom less invasive diagnostic tools, such as high-resolution computerized tomographic angiography (CTA) and magnetic resonance angiography (MRA), were helpful in confirming the diagnosis and in following the evolution of the dissection. CASE PRESENTATION: We present this 51-year old woman who experienced the sudden onset of severe headache without associated neurological deficits. Head CT showed SAH with blood in the interhemispheric fissure, suggesting a ruptured ACA aneurysm. Serial cerebral angiograms failed to demonstrate an aneurysmal sac, but showed evolving irregularities of the ACA consistent with a dissecting aneurysm. These findings were confirmed by CTA and MRA. The patient was treated conservatively and made an excellent recovery. A MRA obtained 2 months later showed slight improvement of the previously visualized ACA dilatation. CONCLUSION: Serial angiograms are often required to confirm the diagnosis and to follow the evolution of an intracranial dissection. With recent advances in neuroradiological techniques, however, critical information can be obtained by less invasive imaging studies, such as CTA and MRA. PMID- 9352813 TI - Aneurysm reinforcement in the anterior circulation. AB - BACKGROUND: Aneurysms are primarily treated with surgical clipping. Unclippable aneurysms are at risk of recurrent bleeding similar to the natural history of ruptured aneurysms. Aneurysm reinforcement with muslin and ethyl 2-cyanoacrylate may provide a safe and useful alternative to reduce the rebleeding rate in those aneurysms found to be unclippable at surgical exploration. METHODS: We examined a population of 19 patients with aneurysms treated by reinforcement with muslin and ethyl 2-cyanoacrylate. One patient was lost to follow-up. Patients had a mean follow-up of 60 months. The yearly risk of rebleeding was calculated and compared to the natural history of the disease. RESULTS: Most of the aneurysms were located in the middle cerebral artery or the anterior communicating artery. The two main causes for reinforcement were a wide base aneurysm or the presence of arterial branches originating from or intimately adherent to the dome. Those patients with aneurysms in which reinforcement was used as the primary and only treatment had a risk of rebleeding of 2.95% per year. The group of patients with ruptured aneurysms in which the reinforcement was used to supplement a clip had no rebleeding. When all the patients with ruptured aneurysms are combined, there is a risk of rebleeding of 1.94% per year. Unruptured aneurysms treated with reinforcement had not shown hemorrhage. No complications related to the use of muslin or ethyl 2-cyanoacrylate were found in this study. CONCLUSION: The reinforcement with muslin and ethyl 2-cyanoacrylate provides some protection in patients with aneurysms for which direct clipping cannot be done and in those clipped cases in which there is a residual neck or dome. This protection is inferior to clipping, but reduces the risk of rebleeding during the first 6 months after the initial hemorrhage. A longer follow-up is needed to determine if it provides better protection for late rebleeding than the natural history of ruptured aneurysms. PMID- 9352814 TI - Volume changes following gamma knife radiosurgery of intracranial tumors. AB - BACKGROUND: The primary goal of radiosurgery for brain tumors is the prevention of further growth. The purpose of this article is to evaluate temporal changes of tumor volume after Gamma Knife radiosurgery on intracranial tumors. METHOD: Some 137 patients with 148 intracranial tumors who were treated with Gamma Knife radiosurgery and underwent radiological follow-up were reviewed. The tumors with high radiosensitivities to conventional external radiation were excluded. RESULT: The median radiological follow-up period was 12 months (range 1.5-38 months). Volume decreased after radiosurgery in 15 of 45 meningiomas; 10 of 37 schwannomas; 6 of 21 pituitary adenomas; 4 of 15 benign gliomas, including both of 2 subependymal giant cell astrocytomas; and 2 of 8 malignant gliomas. Some 87% of meningiomas and 60% of schwannomas whose volume had decreased began to shrink within 12 months and after 12 months, respectively. Transitory increase in volume preceded shrinkage in 16.2% of schwannomas, 13.3% of benign gliomas, 4.8% of pituitary adenomas, and 2.2% of meningiomas. Marked shrinkage occurred in 17 of 19 metastatic tumors and in all 3 neurocytomas shortly after radiosurgery. Of eight malignant gliomas, five began to grow 2-14 months (median = 5 months) after radiosurgery. CONCLUSION: Several points should be considered carefully while following up on patients after radiosurgery: the possibility of transient volume increase, tumor-specific volume change patterns, and the tumor-specific goals of radiosurgery. PMID- 9352815 TI - Clinical features and growth fractions of pituitary adenomas. AB - BACKGROUND: The Ki-67 monoclonal antibody is expressed by proliferating and dividing cells, but not by resting cells. The specificity of the monoclonal antibody, MIB-1, against the Ki-67 antigen has been established by immunostaining of formalin-fixed paraffin-embedded tissue in a microwave oven. METHODS: The growth fraction of 85 pituitary adenomas was studied retrospectively by immunohistochemical analysis using the monoclonal antibody MIB-1. The adenomas were classified into three types: microadenoma, expansive type, and invasive type, based on findings on Gd DTPA enhanced magnetic resonance imaging. RESULTS: The mean MIB-1 index in nonfunctioning microadenomas was higher than in expansive and invasive adenomas, but this difference was not significant. The MIB-1 index in younger patients (under 30 years) with nonfunctioning adenomas was significantly higher than in patients over 40 years of age. One of 14 patients with recurrent disease had an elevated MIB-1 index, but generally patients with an MIB-1 index over 2.0% did not suffer recurrence. The mean MIB-1 index was higher in expansive and invasive functioning adenomas than microadenomas, but not significantly. No correlation between the MIB-1 index and the serum GH or PRL concentration was established. No MIB-1 positive nuclei were observed in two GH producing adenomas treated with the somatostatin analog SMS 201-995. CONCLUSIONS: No significant relationship was identified between growth fraction and the invasiveness or recurrence of pituitary adenomas. The growth fraction of nonfunctioning pituitary adenomas was higher in patients under 30 years than over 40 years of age. PMID- 9352816 TI - Prediction of recurrence in histologically benign meningiomas: proliferating cell nuclear antigen and Ki-67 immunohistochemical study. AB - BACKGROUND: Recurrence in individual patients after complete surgical removal of meningiomas cannot be predicted by histology alone because recurrence occurs even in histologically benign meningiomas. METHODS: We investigated proliferating cell nuclear antigen (PCNA) and Ki-67 labeling indices of histologically benign meningiomas in 95 patients to assess their relationship to recurrence. The labeling index (LI) was expressed as the percentage of tumor cell nuclei immunoreactive for PCNA or Ki-67 to total tumor nuclei counted per section. The cases/specimens comprised the following two groups: (1) nonrecurrent group: 82 specimens from 82 patients without recurrence, (2) recurrent group: 28 specimens from 10 patients with recurrence. RESULTS: Proliferative activities or aggressiveness do not always develop with every recurrence in recurrent meningiomas. The PCNA LI was significantly higher in the recurrent group (3.98% +/- 0.37%) than in the nonrecurrent group (0.71 +/- 0.13%) (p < 0.0001). The Ki 67 LI also was significantly higher in the recurrent group (3.15 +/- 0.40%) than in the nonrecurrent group (0.39 +/- 0.07%) (p < 0.0001). There was a good correlation between the PCNA LI and the Ki-67 LI (coefficient of correlation r = 0.79, p < 0.001). CONCLUSIONS: The results of our study suggested that a PCNA or Ki-67 LI of more than 2% may represent an increased risk for recurrence; therefore, we suggest that radiotherapy or stereotactic radiosurgery should be considered, even for histologically benign meningiomas. PMID- 9352818 TI - Remote effect of brain retraction on regional cerebral blood flow and cerebrovascular reserve on single photon emission computed tomography. AB - BACKGROUND: The purpose of this study is to evaluate the effect of brain retraction 1 year or more after intracranial aneurysm clipping, demonstrated by a regional cerebral blood flow (rCBF) imaging technique. METHODS AND RESULTS: rCBF and cerebrovascular reserve (CV) were evaluated in 40 patients 12-25 months after operation, using single photon emission computed tomography (SPECT) with Tc-99m hexamethylpropylene amine oxime (HMPAO) combined with acetazolamide test. The images were analysed semiquantitatively, focusing on regions of interest (ROIs) chosen for places retracted during the operation. The regions of hypoperfusion in the retracted tissue were clearly visible in 26 cases. Assymmetry of measured activity, expressed in the Assymmetry Index, reached 12% (SEM +/- 8). After injection of acetazolamide during hypercapnia, the assymmetry decreased. CONCLUSIONS: The results confirm the negative role of brain retraction. However, these consequences seem to be diminished by good vasoreactivity. PMID- 9352817 TI - Surgical treatment of epilepsy from schizencephaly with fused lips. AB - BACKGROUND: Surgical treatment of schizencephaly with fused lips has been reported in few cases. In all of the previously reported cases, temporal lobectomy was selected as a major surgical treatment, except for one case with cortical resection. We present a case of direct resection of dysplastic walls of the schizencephalic cleft and the surrounding epileptic area. CASE: This 20-year old college student with medication-resistant epilepsy was surgically treated by subpial cortical resection of the epileptogenic area around a schizencephalic cleft. Magnetic resonance imaging showed an unilateral schizencephalic cleft with fused lips in the right parietal lobe. Pathologic examination demonstrated dysplastic neurons in the epileptogenic cortex. Intraoperative electrocorticography clearly detected epileptiform discharges around the cleft, and the epileptogenic lesion was completely resected. He has been seizure-free for 1 year since the operation and he has no neurologic deficits. CONCLUSION: Subpial resection of the dysplastic cortex surrounding the cleft under the guide of electrocorticography is an effective and minimally invasive procedure for the treatment of schizencephaly. PMID- 9352819 TI - Intraspinal sarcoidosis: diagnosis and management. AB - OBJECTIVE: Isolated intramedullary spinal cord or cauda equina involvement by sarcoidosis is quite rare. We report three patients with intraspinal sarcoidosis and absent systemic manifestations of the disease. The clinical presentation, operative management, electrophysiologic studies, pathology, laboratory investigations, and current therapy are discussed with attention to the previous literature. METHODS: Two of the three patients had a preoperative diagnosis of a cervical intramedullary spinal cord tumor. The third patient had the preoperative diagnosis of an infectious process involving the cauda equina. Magnetic resonance imaging (MRI) with gadolinium did not suggest an inflammatory process. Intraoperative somatosensory evoked potential performed in two patients exhibited normal amplitudes, but a prolonged latency in seven out of eight extremities; with normal central conduction time suggesting a peripheral or radicular involvement. All three patients underwent laminectomy and biopsy of the intraspinal pathology. RESULTS: Pathologic examination demonstrated sarcoidosis in all three patients. Intraoperative observations, intramedullary nodules, and thickening of the meninges were inconsistent with neoplasm and limited the surgical procedure to a biopsy. Frozen sections performed at two of the operations revealed an inflammatory process that confirmed the intraoperative observations. Postoperatively, the diagnostic work-up for all patients was negative for systemic manifestations. CONCLUSIONS: Isolated intraspinal sarcoidosis is a rare process. The current management for intramedullary spinal cord or cauda equina sarcoidosis is prolonged corticosteroids. The surgeon should not attempt complete resection if this granulomatous process is suspected. PMID- 9352820 TI - Multiple intramedullary spinal sarcoidosis: case report. AB - BACKGROUND: According to past reported cases, spinal intramedullary sarcoidosis has the radiologic characteristics of a single enhanced lesion with cord swelling. However, it is difficult to make a diagnosis using only radiologic examinations. Including our case, 20 cases have been reported of intramedullary spinal sarcoidosis. Only six cases were diagnosed as primary spinal sarcoidosis. We present a rare case of multiple spinal intramedullary sarcoidosis without cord swelling. CASE DESCRIPTION: This 63 year-old man was afflicted with progressive paraparesis and numbness of the lower extremities. Magnetic resonance imaging showed intramedullary lesions without cord swelling. The patient underwent biopsy for diagnosis and the specimen showed a noncaseating granuloma mainly composed of epitheloid cells. Postoperative corticosteroid therapy was effective and the lesion disappeared 1 month after the operation. CONCLUSIONS: Biopsy is a less invasive method and is useful for early diagnosis. Early diagnosis is important for spinal sarcoidosis so that high-dose corticosteroid therapy can be commenced while the lesion still has a good chance of being successfully treated. PMID- 9352821 TI - Neurosarcoid infiltration of the ventricular catheter causing shunt failure: a case report. AB - BACKGROUND: Neurosarcoid is known to develop in 5% of patients with sarcoidosis. A frequent manifestation of this condition is hydrocephalus, which will often require treatment with a ventricular shunt. METHODS: Presented here is the case of a patient whose initial manifestation of neurosarcoidosis was hydrocephalus, and who then represented with multiple shunt failures. RESULTS: On two revisions, the proximal catheter, after removal, was found to be occluded with noncaseating granulomatous material that had infiltrated the shunt lumen. CONCLUSIONS: Although shunts may become occluded by inspissated proteinaceous or cellular debris, the occlusion of a shunt catheter by the ingrowth of the noncaseating granulomatous material of neurosarcoid has not been described before. This patient's course, the histopathologic findings, and neurosarcoidosis in general are discussed here. PMID- 9352822 TI - MRI for CSF rhinorrhea. PMID- 9352823 TI - Choice of a tone-pip envelope for frequency-specific threshold evaluations by means of the middle-latency response: normally hearing subjects and slope of sensorineural hearing loss. AB - The effects of stimulus rise-fall and plateau times on the middle-latency response (MLR) waveform (Na-Pa amplitude and Pa latency) were investigated in 14 normally hearing subjects and an objective MLR threshold was evaluated at low and middle frequencies in ten normally hearing subjects and ten patients with slope of sensorineural hearing loss, using a selected stimulus-envelope time. After analyzing the effects of envelope times on the MLR waveform and the spectra of tone-pips, it was found that a rise-fall time of 4 ms with a plateau of 2 ms (4-2 4) is an acceptable compromise between a synchronous discharge and frequency specificity for estimating the MLR threshold. The MLR threshold produced by 4-2-4 tone-pips approximated the psychoacoustic threshold at low and middle frequencies in the normal and hearing impaired subjects. This demonstrates the clinical usefulness of the MLR in estimating low- and middle-frequency thresholds. PMID- 9352824 TI - Radiation tolerance of the cochlear nerve at the gamma-knife in rabbits. AB - Since the first treatment of acoustic neurinoma using the gamma-knife by Leksell, a series of cases have been reported with good control rates. However, the most frequent complication is delayed hearing loss which occurs in more than 50% of patients. The purpose of this study was to define a safe dose by analyzing the radiosurgical dose-response relationship and histological effects on the normal cochlear nerve in rabbit. The rabbits had computed tomography (CT)-guided stereotactic radiosurgery on their cochlear nerves in the internal auditory canal with a 4 mm collimator focusing of a gamma-unit. Maximum doses of 10, 20, 30, 40, 60, 80, 100, 200 and 500 Gy were administered. After the radiosurgery, auditory brain stem responses (ABR) and the behavior of the rabbits were evaluated periodically. At the conclusion, histological investigations were performed. No physiological or histological findings were observed from doses of 30 Gy or below during the 12 month period after the radiosurgery. A dose of 100 Gy caused a severe ABR threshold elevation, vestibular dysfunction and facial palsy. Necrosis and demyelination of nerves were observed pathologically. In this study, we determined that the safe dose to the normal cochlear nerve during radiosurgery was under 40 Gy in rabbits, and complications seemed to vary due to individual differences in radiation tolerance. PMID- 9352825 TI - The effect of intravenous lidocaine injection on hearing thresholds. AB - The effect of intravenous injection of lidocaine on hearing thresholds was studied in normal subjects. Continuous and intermittent tones at 1, 4 and 8 kHz were used as stimuli and the threshold change with lidocaine injection was measured using a self-recording audiometer (Bekesy audiometer). Both increases and decreases in the threshold were observed. The former occurred more frequently than the latter. In cases of a threshold increase, lidocaine injection exhibited a frequency specific effect; the higher the frequency, the more often the threshold was increased by lidocaine injection. There was no significant difference in threshold changes between continuous and intermittent tones. The present results suggest that lidocaine may act on the inner ear hair cells. PMID- 9352826 TI - A case report of fluctuant sensorineural hearing loss after hepatitis B vaccination. AB - Hepatitis B vaccine is known to induce accidents and side-effects. We report a case of unilateral fluctuant sensorineural hearing loss with tinnitus after a series of hepatitis B vaccinations. Tinnitus regressed and hearing thresholds were normalized 6 months later. The authors discuss the potential causality of hepatitis B vaccine in the occurrence of fluctuant sensorineural hearing loss and propose a physiopathological hypothesis to explain this unusual case. PMID- 9352827 TI - Cochlear implant after reconstruction of the external bony canal wall and tympanic cavity in radically mastoidectomized patients with cholesteatoma. AB - One of the postoperative complications of cochlear implants in patients, who previously received radical mastoidectomy, is an exposure of electrode by breakdown of thin epithelium in the open mastoid cavity. To avoid such complications, in the first stage, radical mastoidectomy with the reconstruction of the posterior bony canal wall and mastoid obliteration with bone chips and plates and the creation of the new tympanic cavity, were performed. One or 3 years later, implantation of a 22-channel cochlear implant, as the second stage procedure, was successfully performed in three patients with profound sensorineural hearing loss, due to cholesteatoma in the side of the ear in which cochlear implantation was indicated. The advantages of this technique are as follows: (1) Electrode is protected from the cavity problems, such as chronic infection or erosion of the epithelium in the open mastoid cavity; and (2) reconstruction of the new tympanic cavity and tympanic membrane is beneficial for avoidance of electrode exposure in the mastoid and tympanic cavity. PMID- 9352828 TI - Comparison of TEOAE with Play audiometry for screening hearing problems in children. AB - To evaluate the usefulness of transiently evoked otoacoustic emissions (TEOAEs) for hearing screening of children at around 3 years of age, measurements were done together with Peep show test in a group of 47 children (n = 93 ears). A stimulus sound of 30 dB nHL was used as the screening intensity for the TEOAE measurement. All measurements were done with awake subjects. Twenty seven ears, all of which were revealed to have normal hearing (within 20 dB HL, assessed by Peep show test) and tympanograms, showed positive TEOAE results. Furthermore, TEOAEs were sensitive to the presence of middle ear conductive impairment, showing negative results. We conclude that, compared with Play audiometry, TEOAE measurements can not yield quantitative results, but can yield qualitative results for determining the presence of hearing impairment without sedative induced sleeping in this critical age of children. PMID- 9352829 TI - Distribution of nitric oxide in the nasal mucosa of the rat: a histochemical study. AB - We evaluated the distribution of nitric oxide (NO) in the rat nasal mucosa using nicotineamide adenosine dinucleotide phosphate (NADPH)-diaphorase histochemistry. The NADPH-diaphorase positive nerve fibers in the nasal mucosa were observed around blood vessels and submucosal glands and in sphenopalatine ganglions. Strong positive reactions for NADPH-diaphorase were observed in ganglions as compared with the other tissues. In septal and turbinate mucosa, positive reactions for NADPH-diaphorase were mainly seen in the anterior portion, and a few positive reactions were observed in the posterior portion. No positive reactions for NADPH-diaphorase were demonstrated in the sinus mucosa. These results suggest that NO may be related to regulation of blood flow, glandular secretion and neurotransmission, and also that NO may play an important role in the defense mechanism of the upper airway system against external environments. PMID- 9352830 TI - Use of vertical median forehead flap in the reconstruction of the anterior skull base: report of two cases. AB - Improvements in reconstruction of the skull base have made craniofacial surgery safe. Reconstruction of the anterior skull base must provide a seal between the cranial cavity and upper respiratory tract, as well as offer structural support for the brain. A wide variety of local flaps have been designed. The choice of flap in individual cases depends on the location and size of the defect. We report a reconstructive technique for the anterior skull base with vertical median forehead flaps which we used to treat two patients, one patient with adenocarcinoma and the other with leiomyosarcoma. Both were lesions of the ethmoid sinuses and nasal cavity. PMID- 9352831 TI - A new nutritional assessment in patients with oral and maxillofacial malignancies. AB - Combined subjective and objective nutritional assessment was performed on admission in 127 patients with oral and maxillofacial malignancies. On the basis of the nutritional assessment result, three typical nutritional parameters-body weight (BW) (X1), mid-upper-arm circumference (MAC) (X2), and hand grip strength (HGS) (X3) were used to establish a new nutritional assessment method that was developed by a computer-based discriminant analysis. The established model was as follows: Y1 = -126 + 1.09X1 + 1.34X2 + 0.23X3; Y2 = -95.63 + 0.96X1 + 1.17X2 + 0.19X3. In the model Y1 was regarded as good nutrition and Y2 as malnutrition. The larger value of Y stood for the patient's nutritional status (i.e. Y1 > Y2, well nourished; Y1 < Y2 malnourished). The new nutritional assessment correlates well with the combined subjective and objective nutritional assessment with a total agreement rate of 88.2%. With its simplicity and accuracy, the new nutritional assessment deserves a wide application in clinical situation. PMID- 9352832 TI - The significance of arytenoid edema following radiotherapy of laryngeal carcinoma with respect to residual and recurrent tumour. AB - We sometimes experience patients with persistent or progressive arytenoid edema, among which residual or recurrent cancer is often accompanied. Because it is difficult to distinguish tumour rest or recurrence from normal tissue sequelae in the early period after irradiation, it is important to know both the contributing factors for arytenoid edema, and the incidence of residual or recurrent tumours in patients with postirradiation laryngeal edema. We therefore reviewed the charts of 67 patients with early laryngeal carcinoma who had received a curative dose of irradiation in the last 5 years. Fourteen patients (20.9%) had moderate or severe laryngeal edema persisting for or developing at more than 3 months after completion of a course of definitive radiotherapy. The incidence was highest in supraglottic T2 disease, followed by glottic T2 tumour. Of the 14 patients with edema, six (42.9%) had persistent or recurrent disease. The primary disease was uncontrolled in 18 patients, 17 of whom received successful salvage surgery. In patients without residual tumours, the edema was usually moderate and resolved within a year, although four patients had chronic edema lasting more than a year after treatment. All four had supraglottic T2 lesions and received 70 Gy of X-ray. We also reviewed, for sake of comparison, the records of 38 patients treated with radiotherapy at doses of more than 40 Gy between 1975 and 1980, when endoscopic microsurgery for laryngeal cancer was introduced as a primary part of treatment. The incidence of persistent or late developed edema over the period, though not significant, was 36.8%: nearly twice that of the last 5 years. Microscopic endolaryngeal surgical procedures seem to have been a causal factor for edema in this period. PMID- 9352833 TI - Correlation between anthropometric measurements of the oropharyngeal area and severity of apnea in patients with snoring and obstructive sleep apnea. AB - The aim of this study was to investigate the relationship between severity of apnea and anthropometric oropharyngeal measurements in patients with snoring and obstructive sleep apnea. A total of 22 patients complaining of snoring and apneic spell during sleep were evaluated by polysomnographic and anthropometric measurements of the oropharyngeal area. The horizontal width of the uvula at the mid-point and the length of the uvula were measured using a T-shaped ruler. The distance between the anterior pillars, posterior pillars and retromolar raphes were also measured. The correlation between these anthropometric measurements and polysomnographic parameters including the respiratory disturbance index (RDI) and the lowest arterial O2 saturation level (lowest SaO2) of the patients were analyzed. Of the anthropometric measurements, the horizontal width of the uvula showed a significant correlation with RDI and lowest SaO2. The results of the present study indicate that patients with broader uvula may have severer sleep apnea and that anthropometric oropharyngeal measurements may give additional information to polysomnographic findings for selecting surgical candidates. PMID- 9352834 TI - Radiological findings of adenolymphoma (Warthin's tumor). AB - The radiological findings of adenolymphomas (Warthin's tumor) treated in six hospitals between 1985 and 1994 were compared with the operative and histological findings and the usefulness of the radiological examinations was evaluated. The total number of patients was 72. The mean age was 61.8 years; 61 were males and 11 were females. All tumors developed in the parotid gland. Tc-99m-pertechnetate salivary gland scanning was performed in 13 patients and an increased uptake of the isotope was observed in only six patients. Even if Tc-99m-pertechnetate salivary gland scanning does not reveal intense accumulation, this tumor should not be ruled out. By computed tomography (CT), ultrasonography (US), and magnetic resonance imaging (MRI), the margin of all tumors was evident; however, the contents of the tumor varied. The contents and multiplicity of the tumors were well demonstrated by MRI, which was found to be the most accurate imaging modality. PMID- 9352835 TI - Clinical study of esophageal foreign bodies attributable to PTP material. AB - Retrospective survey of patients with esophageal foreign bodies who were treated in the Osaka Medical College over the past 21 years, and the patients with esophageal foreign bodies attributed to press through package (PTP) who were treated at the Osaka Central Emergency Clinic, a representative holiday emergency institution in Japan, was carried out. The incidence of foreign bodies attributed to PTP material tends to increase throughout the period reviewed. The patients over 60 years of age accounted for 25/28 (89%) of all PTP patients in Osaka Medical College and 27/32 (84%) in Osaka Central Emergency Clinic, strongly suggesting that PTP dysphagia is most common in elderly patients. Diagnostic methods and preventive measures against PTP foreign bodies were discussed. PMID- 9352836 TI - Basaloid squamous carcinoma of the larynx: report of a case. AB - Basaloid squamous carcinoma (BSC) is a rare neoplasm. We present a case of basaloid squamous carcinoma of the larynx in a 57-year-old male patient. The diagnosis before treatment was supraglottic carcinoma (T3N1MO) and biopsy of the larynx revealed a poorly differentiated squamous cell carcinoma. Total laryngectomy and right radical neck dissection were performed, and pathological studies of a specimen removed from the larynx revealed BSC of the larynx. The patient's postoperative progress was uneventful, however, 12 months later he developed lung metastasis of the left side. The patient underwent partial resection of the lung. He developed recurrence of lung metastasis 6 months later. Chemotherapy with cisplatin (CDDP) and vindesine sulfate (VSD) was administered in two courses, but the efficacy was evaluated as no change (NC). At present, 26 months after the first visit, he has been asymptomatic with lung metastasis, and there was no evidence of recurrence in the neck. PMID- 9352840 TI - The pros and cons of Canada's government-run system. PMID- 9352838 TI - New paradigm for inherited colon cancer. PMID- 9352837 TI - Small cell carcinoma of the larynx: imaging findings. AB - Small cell carcinoma of the larynx is an uncommon epithelial tumor, which is the most aggressive subtype of neuroendocrine carcinomas. Because of its nonspecific clinical and radiological manifestations, the diagnosis of small cell carcinoma of the larynx is essentially based on the light microscopic examination aided by electron microscopy or immunohistochemical staining. We report a case of supraglottic small cell carcinoma accompanied by large bilateral cervical lymph node metastasis ocurring in a 70-year-old man. On CT scans, no area of low attenuation indicating necrosis was demonstrated within such large metastatic lymph nodes. We suggest that small cell carcinoma of the larynx should be included in the diagnostic considerations when a laryngeal mass is accompanied by large cervical lymph nodes without necrosis shown by CT. PMID- 9352839 TI - Opening the intestinal gate to Peyer's patches. PMID- 9352841 TI - History of the mouse in gastrointestinal research: part I. PMID- 9352842 TI - Image of the month. Disseminated infections, especially candidiasis in immunocompromised individuals, cat-scratch fever, and infiltrative diseases such as leukemia or lymphoma. PMID- 9352843 TI - Localization of the Bannayan-Riley-Ruvalcaba syndrome gene to chromosome 10q23. AB - BACKGROUND & AIMS: Bannayan-Riley-Ruvalcaba syndrome is a congenital syndrome with characteristic features of macrocephaly, cognitive and motor dysfunction, subcutaneous and visceral lipomas and hemangiomas, and intestinal juvenile polyposis. It has been suggested that Bannayan-Riley-Ruvalcaba syndrome may be a variant of juvenile polyposis coli because of the shared features of intestinal juvenile polyps. The aim of this study was to precisely map loss of DNA from 2 patients with intestinal juvenile polyposis and karyotypic abnormalities involving chromosome 10q. METHODS: DNA was extracted from peripheral leukocytes drawn from each patient and each patient's biological parents. The DNA was amplified by polymerase chain reaction using primers specific for microsatellites located on chromosome 10q. RESULTS: Precise mapping localized a maximal distance of 1.0 cM that was commonly deleted from each patient's genome, between D10S541 and D10S1735. This area overlaps the region for Cowden disease, a distinct hamartomatous intestinal polyposis syndrome with increased risk of breast and thyroid carcinoma. CONCLUSIONS: The three hamartomatous polyposis syndromes, Bannayan-Riley-Ruvalcaba syndrome, juvenile polyposis coli, and Cowden disease, may share the same genetic defect because of their common map localization to chromosome 10q23. PMID- 9352844 TI - Osmodependent dynamic localization of the multidrug resistance protein 2 in the rat hepatocyte canalicular membrane. AB - BACKGROUND & AIMS: Circumstantial evidence suggests a regulation of biliary secretion by transporter insertion and retrieval into and from the canalicular membrane. This study was undertaken to provide direct evidence for such a process. METHODS: Osmosensitivity of the subcellular localization of the mrp2 gene-encoded conjugate export pump (MRP2) was studied by immunofluorescence and confocal laser scanning microscopy of isolated hepatocyte aggregates and in perfused rat liver. RESULTS: MRP2 was localized largely in membranes of the pseudocanaliculi formed by isolated hepatocyte aggregates during hypo-osmotic exposure, whereas after hyperosmotic exposure MRP2 was also detectable in intracellular vesicles. In perfused liver, the EAG15 antibody specific for rat MRP2 and the ZO-1 antibody specific for tight junctions produced immunostaining of the canalicular membrane. However, the relative amount of MRP2 increased significantly in the pericanalicular region with increasing perfusate osmolarity, as shown by confocal microscopy of intracellular vesicles containing MRP2 (but not ZO-1) and by computed densitometry. The osmodependent distribution of MRP2 between the canalicular membrane and intracellular, pericanalicular vesicles occurred within 30 minutes and was fully reversible. CONCLUSIONS: The findings provide direct evidence for an osmosensitive dynamic insertion and retrieval of the canalicular MRP2 transporter into and out of the canalicular membrane. PMID- 9352845 TI - Physician specialty and variations in the cost of treating patients with acute upper gastrointestinal bleeding. AB - BACKGROUND & AIMS: Upper gastrointestinal tract bleeding is a frequent cause of hospitalization. The goal of this study was to assess whether the cost of treating patients with upper gastrointestinal bleeding varies among surgeons, internists, and gastroenterologists. METHODS: A retrospective study of 124 patients admitted with acute upper gastrointestinal hemorrhage was performed. Patients were stratified into three groups based on a validated risk score; length of stay and hospital costs were compared among patients primarily cared for by internists, surgeons, and gastroenterologists. RESULTS: The median length of stay (2 days) for patients admitted to the gastroenterology service was significantly shorter than for patients admitted under the care of other physicians (P < 0.05). The median hospitalization cost ($2856) for patients admitted to the gastroenterology service was significantly lower than for patients admitted to the other services (P < 0.01). There were no significant differences in the time to endoscopy among services. CONCLUSIONS: Patients admitted to an urban teaching hospital directly under the care of a gastroenterologist had shorter hospital stays that were significantly less costly than patients under the primary care of internists or surgeons. The difference in length of stay reflects the time interval between endoscopy and discharge. PMID- 9352846 TI - Familial aggregation of gastroesophageal reflux in patients with Barrett's esophagus and esophageal adenocarcinoma. AB - BACKGROUND & AIMS: Barrett's esophagus and adenocarcinoma are complications of gastroesophageal reflux disease. The aim of this study was to look for evidence of a familial predisposition to reflux. METHODS: Index patients with adenocarcinoma (n = 27), Barrett's esophagus (n = 40), and reflux esophagitis (n = 55) were recruited from tertiary care and community populations. Parents and siblings of patients (n = 243) and their spouses' relatives (n = 230) completed reflux symptom questionnaires (response rate, 86%). RESULTS: Reflux symptoms were significantly more prevalent among parents and siblings of patients with adenocarcinoma (43% vs. 23%) and Barrett's esophagus (46% vs. 27%) than spouse control relatives. No significant difference was found for the reflux esophagitis group (33% vs. 29%). Reflux was more prevalent in siblings than spouses of patients with Barrett's esophagus (41% vs. 12%) and adenocarcinoma (40% vs. 6%), a difference that was not found with reflux esophagitis (24% vs. 32%). Reflux was associated with obesity, 41% vs. 28% in the nonobese; smoking, 45% vs. 31% in nonsmokers; and men, 39% vs. 27% in women. CONCLUSIONS: There may be a genetic predisposition to the development of reflux in families of patients with Barrett's esophagus and esophageal adenocarcinoma. For uncomplicated reflux esophagitis, environmental factors appear more important. PMID- 9352847 TI - Impaired deglutitive airway protection: a videofluoroscopic analysis of severity and mechanism. AB - BACKGROUND & AIMS: Laryngeal vestibule penetration is a prerequisite for deglutitive aspiration. This study aimed to analyze the mechanism and model the risk of laryngeal penetration before or during the pharyngeal swallow. METHODS: Videofluoroscopic swallowing studies of 29 patients with neurogenic dysphagia with penetration before or during the pharyngeal swallow were compared with 12 controls. A stepwise regression analysis was used to define the coordinative defects leading to bolus penetration into the laryngeal vestibule. The mechanism was biomechanically analyzed. RESULTS: The stepwise regression modeled a laryngeal penetration index from the coordination between laryngeal vestibule closure and bolus release at the glossopalatal junction and the timing of upper esophageal sphincter opening relative to glossopalatal junction opening. The model accounted for 86% of the observed variance in severity of laryngeal penetration among the dysphagics. The observed incoordination resulted from both delayed initiation and slowed enactment of deglutitive laryngeal elevation. CONCLUSIONS: A dysphagic individual's risk of incurring laryngeal penetration before or during 1-, 3-, or 5-mL swallows is proportional to two temporal measures of coordination made from 1-mL swallows. The severity of the relevant defects (delayed and slowed laryngeal elevation) is proportional to the severity of swallow dysfunction. PMID- 9352848 TI - Mesalamine in the maintenance treatment of Crohn's disease: a meta-analysis adjusted for confounding variables. AB - BACKGROUND & AIMS: The benefit of mesalamine for maintenance of remission in Crohn's disease is controversial. The aim of this study was to assess the effectiveness of mesalamine in maintaining remission of quiescent Crohn's disease. METHODS: Pertinent randomized clinical trials were selected using MEDLINE (1986-1997) database, reference lists from published articles or reviews. Fifteen randomized, controlled trials of mesalamine maintenance therapy involving a total of 2097 patients were selected. The crude rates of patients with symptomatic relapse in treated and control groups were extracted according to the intention-to-treat method. RESULTS: Therapy with mesalamine significantly reduced the risk of symptomatic relapse (pooled risk difference, -6.3%; 95% confidence interval, -10.4% to -2.1%). The pooled risk difference was significant in the postsurgical setting (-13.1%; 95% confidence interval, -21.8% to -4.5%) but not in the medical setting (-4.7%; 95% confidence interval, -9.6% to 2.8%). Multivariate model predicts that the probability of symptomatic relapse significantly decreases with mesalamine treatment, by increasing proportion of patients with ileal disease, with prolonged disease duration, and with surgically induced remission. CONCLUSIONS: Mesalamine may be recommended for maintaining remission of quiescent Crohn's disease. The benefit is mainly observed in the postsurgical setting, in patients with ileitis and with prolonged disease duration. PMID- 9352849 TI - Effects of mesalamine on the hsp72 stress response in rat IEC-18 intestinal epithelial cells. AB - BACKGROUND & AIMS: Mesalamine has many effects and is commonly used for the treatment of inflammatory bowel diseases. Because sodium salicylate, a related compound, modulates the heat shock protein (hsp72) response in nonepithelial cells, the possibility that mesalamine confers cell protection by increasing intestinal epithelial hsp72 expression was examined. METHODS: Rat intestinal IEC 18 cells were treated with 0.3-3 mmol/L mesalamine and thermally stressed (39 degrees C-42 degrees C) for 23 minutes. The effects of mesalamine on basal expression and the threshold and time course of hsp72 thermal induction were determined. RESULTS: Although mesalamine had no effects on the basal hsp72 expression or its thermal activation threshold in IEC-18 cells, it accelerated and augmented thermal induction of hsp72 within the first 2 hours of exposure. This was associated with a transient increase in heat shock factor-heat shock element binding and enhanced cellular protection against oxidant-induced injury. In contrast, both mesalamine and sodium salicylate have been shown to lower the thermal induction threshold but not to enhance the hsp72 response in HeLa cells. CONCLUSIONS: Mesalamine augments thermal induction of the intestinal epithelial hsp72 expression in a manner that differs from that in nonintestinal epithelial cells. This effect is accompanied by increased cellular protection against oxidant injury. PMID- 9352850 TI - Peroxynitrite-induced apoptosis in human intestinal epithelial cells is attenuated by mesalamine. AB - BACKGROUND & AIMS: Peroxynitrite (PN), a potent oxidant, has been implicated in the pathogenesis of gut inflammation and epithelial cell apoptosis. The aim of this study was to investigate mesalamine, a standard therapy for inflammatory bowel disease, to see if it attenuates PN-induced cytotoxicity in human intestinal epithelial cells and if mesalamine directly interacts with PN or its precursor, nitric oxide. METHODS: T84 and HT29 cells were divided in several protocols: mesalamine was administered 2 hours before, simultaneously, or 30 minutes after PN. T84 cells, grown in filter chamber inserts, were used to determine if basolateral or apical administration of PN initiated apoptosis and epithelial barrier function. The effects of mesalamine on PN decomposition and NO half-life were determined. RESULTS: Mesalamine resulted in a dose-dependent decrease in PN-induced apoptosis, whether mesalamine was administered before (>10 micromol/L; IC50, 16 micromol/L), simultaneously (25-200 micromol/L; IC50, 24 micromol/L), or 30 minutes (200 micromol/L) after PN exposure. Mesalamine protected the epithelial barrier function of T84 cells against PN. Mesalamine rapidly degraded PN, whereas the half-life of NO was not affected. CONCLUSIONS: The beneficial effects of mesalamine in the treatment of inflammatory bowel disease may involve an attenuation of PN-induced cell injury and apoptosis through direct and indirect mechanisms without affecting NO levels. PMID- 9352851 TI - Neutrophil migration across model intestinal epithelia: monolayer disruption and subsequent events in epithelial repair. AB - BACKGROUND & AIMS: Acute inflammation of the intestine is associated with transepithelial migration of polymorphonuclear leukocytes (PMNs) and epithelial wounds that rapidly reseal. The aim of this study was to determine mechanisms by which such PMN-induced epithelial wounds reseal. METHODS: Epithelial wound closure was modeled in vitro using T84 intestinal epithelial cells and PMNs. Wound closure was analyzed by confocal microscopy and by determination of barrier function. Wounds were highlighted by apical labeling with antibody to a basolaterally restricted ligand, beta1-integrin. RESULTS: High-density PMN transepithelial migration for 70-110 minutes produced multifocal epithelial wounds that were 1-120 microm in diameter and markedly diminished epithelial barrier function that returned to baseline within 12-20 hours. Large wound closure was initiated by cell flattening and extension of F actin/vinculin/paxillin-enriched lamellipodia at the leading edge. As wounds became small (approximately <30 microm), epithelial cells at the wound edges assumed columnar phenotype with poorly formed or absent lamellipodia. Apical localized circumferential, dense F-actin/myosin II rings were found to encircle such wounds, suggesting final closure by a sphincter-like contraction. CONCLUSIONS: These data model mucosal repair in acute inflammatory conditions and, for the first time, show sequential early and late mechanisms by which epithelial discontinuities repair. PMID- 9352853 TI - Relation between integrin alpha7Bbeta1 expression in human intestinal cells and enterocytic differentiation. AB - BACKGROUND & AIMS: Cell-laminin interactions are principally mediated by specific membrane receptors of the integrin family. The integrin alpha7beta1 is one of them. Its expression in the intestine has not yet been investigated although it appears to be a key element in muscle cell differentiation. In this study, the expression of its three known isoforms has been analyzed in developing and adult small intestine and in intestinal cell lines. METHODS: The expression of the integrin alpha7beta1 was analyzed by indirect immunofluorescence, Western blotting, immunoprecipitation, and reverse-transcription polymerase chain reaction. RESULTS: The alpha7B isoform, but not the alpha7A and C isoforms, was detected in intestinal epithelial cells. In vivo, the presence of the alpha7B subunit was closely paralleled with (1) acquisition of differentiation characteristics during development and along the crypt-villus axis in the adult small intestine and (2) loss of enterocytic functions in the re-differentiated colonic epithelium. In vitro, the expression of alpha7B was also shown to correlate with the acquisition of enterocytic functions. In Caco-2 cells, the alpha7Bbeta1 integrin was found transiently up-regulated at the onset of sucrase isomaltase expression. CONCLUSIONS: Taken together, these results suggest that alpha7Bbeta1 expression is correlated with human intestinal cell differentiation. PMID- 9352852 TI - Apical expression of functional asialoglycoprotein receptor in the human intestinal cell line HT-29. AB - BACKGROUND & AIMS: The asialoglycoprotein receptor localizes to the basolateral membrane of hepatocytes and to the apical membrane of enterocytes. The aim of this study was to examine HT-29 cells as a polarized cell model for studying apically localized endogenous asialoglycoprotein receptor. METHODS: Subunits H1 and H2 (human) were detected by Western blot and immunoprecipitated using subunit specific antisera against hepatic receptor peptides. Receptor function was assessed by uptake of iodinated asialo-orosomucoid, immunoglobulin (Ig) A1, and haptocorrin. Immunocytochemistry was analyzed by standard light and confocal microscopy. RESULTS: Receptor content of the minor subunit, H2, was predominant. HT-29 cells mediated specific uptake and degradation of 125I-asialo-orosomucoid. A high-affinity (0.6 x 10(-9) mmol/L) and a low-affinity binding site were present. The specific ligand binding capacity of the apical surface was approximately twice that of the basolateral surface. Immunocytochemistry revealed a predominant apical membrane location of the minor receptor subunit, with some intracellular receptor. The apical H2 subunit was preferentially labeled with amino acid precursors compared with basolaterally located subunit. Human IgA1 bound specifically to HT-29 cells with a molar ratio of 0.26 compared with asialo orosomucoid; porcine haptocorrin bound with a molar ratio of 1.35. CONCLUSIONS: HT-29 cells produce a functional apically located asialoglycoprotein receptor and provide a model for receptor trafficking in the enterocyte. PMID- 9352854 TI - Purinergic fast excitatory postsynaptic potentials in myenteric neurons of guinea pig: distribution and pharmacology. AB - BACKGROUND & AIMS: Adenosine triphosphate (ATP) acting at P2 receptors mediates some fast excitatory postsynaptic potentials (fEPSPs) in myenteric neurons of guinea pig ileum. The present studies investigate the distribution of purinergic fEPSPs along the length of the gut and characterize the P2-receptor subtype mediating fEPSPs. METHODS: Conventional intracellular electrophysiological methods were used to record from myenteric neurons in vitro. RESULTS: At a membrane potential of -97 +/- 1 mV, the amplitude (25 +/- 1 mV; n = 307) of fEPSPs was similar along the gut. Hexamethonium (100 micromol/L) inhibited fEPSPs in the gastric corpus by 98% +/- 1% (n = 31) and in the duodenum, ileum, taenia coli, proximal colon, and distal colon by 42%-55%. In the presence of hexamethonium, suramin (100 micromol/L) or the P2X antagonist pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid (PPADS, 10 micromol/L) reduced the control fEPSP amplitude in the duodenum, ileum, taenia coli, proximal colon, and distal colon by 71%-84%. The pharmacology of the purinergic fEPSPs was investigated in detail in the ileum. Noncholinergic fEPSPs were concentration-dependently (1-30 micromol/L) inhibited by PPADS (50%-inhibitory concentration, 3 micromol/L). In addition, alpha,beta-methylene 5'-adenosine triphosphate (1 micromol/L) also reduced purinergic fEPSPs. CONCLUSIONS: Fast EPSPs mediated in part through P2X receptors are prominent in myenteric neurons along the small and large intestines but are rare in the gastric corpus. PMID- 9352855 TI - Impaired expression of nitric oxide synthase in the gastric myenteric plexus of spontaneously diabetic rats. AB - BACKGROUND & AIMS: The mechanism responsible for defective gastric accommodation in diabetes is unknown. The aim of this study was to investigate if this abnormality is due to a defective nitric oxide pathway secondary to impaired nitric oxide synthase (NOS) expression in the gastric myenteric plexus. METHODS: To test this hypothesis, we studied nonadrenergic, noncholinergic (NANC) relaxation, NOS activity, NADPH diaphorase histochemistry, NOS immunohistochemistry, NOS immunoblotting, and NOS messenger RNA expression in the gastric myenteric plexus of spontaneously diabetic biobreeding/Worcester (BB/W) rats. Age-matched nondiabetic Wistar rats were used as controls. RESULTS: Gastric neuromuscular preparations from control rats showed a frequency-dependent NANC relaxation in response to transmural stimulation. This relaxation was markedly antagonized by N(G)-nitro-L-arginine-methyl ester, indicating mediation by the neuronal release of NO. The NANC relaxation in gastric muscle preparations obtained from diabetic BB/W rats was significantly impaired. The number of NOS immunoreactive cells in the gastric myenteric plexus and the NOS activity were significantly reduced in diabetic BB/W rats, suggesting that NOS synthesis is impaired in diabetes. Northern blot analysis showed that the density of NOS messenger RNA bands at 9.5 kilobases was significantly reduced in the gastric tissues of diabetic BB/W rats. CONCLUSIONS: These results indicate that gastric relaxation in diabetics is hampered mainly by impaired NOS expression in the gastric myenteric plexus. PMID- 9352856 TI - Histamine, acting via H3 receptors, inhibits somatostatin and stimulates acid secretion in isolated mouse stomach. AB - BACKGROUND & AIMS: The role of histamine H3 receptors in the regulation of gastric acid secretion is unclear. The present study was designed to characterize the location of H3 receptors in the fundus of the stomach and the mechanism by which these receptors regulate acid secretion. METHODS: Acid, somatostatin, and histamine secretions were measured in the isolated mouse stomach. RESULTS: Thioperamide (H3 antagonist) increased somatostatin and decreased histamine and acid secretion in a concentration-dependent manner. (r)-alpha-Methylhistamine (H3 agonist) had the opposite effect, decreasing somatostatin and increasing histamine and acid secretion. The pattern implies that endogenous histamine, acting via H3 receptors, exerts an inhibitory paracrine influence on somatostatin secretion. Somatostatin antibody increased basal histamine secretion and abolished the decrease in histamine and acid secretion induced by thioperamide, confirming that changes in histamine and acid secretion induced by the activation of H3 receptors reflected changes in somatostatin secretion. Similar effects were obtained when acid secretion was stimulated by histamine: thioperamide augmented somatostatin and thus inhibited acid secretion, and (r)-alpha-methylhistamine attenuated somatostatin and increased acid secretion. CONCLUSIONS: Reciprocal inhibitory paracrine pathways link histamine and somatostatin cells in the gastric fundus. Histamine, acting via H3 receptors, augments acid secretion by eliminating the inhibitory influence of somatostatin. PMID- 9352857 TI - Roles of prostaglandin E-receptor subtypes in gastric and duodenal bicarbonate secretion in rats. AB - BACKGROUND & AIMS: Receptors activated by prostaglandin (PG) E2 are pharmacologically subdivided into four subtypes (EP1-EP4). The EP-receptor subtype(s) involved in stimulation of gastroduodenal HCO3- secretion in rats were investigated. METHODS: Under urethane anesthesia, a stomach mounted in an ex vivo chamber or a proximal duodenal loop was perfused with saline, and HCO3- secretion was measured using a pH-stat method. RESULTS: Intravenous PGE2 increased HCO3- secretion by the gastroduodenal mucosa; this action was verapamil sensitive and, only in the duodenum, potentiated by isobutylmethyl xanthine (IBMX). Duodenal HCO3- secretion was stimulated by enprostil, sulprostone (EP1/EP3 agonist), misoprostol (EP2/EP3 agonist), and ONO-NT012 (EP3 agonist) but was not affected by butaprost (EP2 agonist) or 17-phenyl-PGE2 (EP1 agonist). Gastric HCO3- secretion was stimulated by sulprostone, enprostil, and 17-phenyl-PGE2 but not by misoprostol, butaprost, or ONO-NT012. SC-51089 (EP1 antagonist) inhibited the HCO3--stimulatory action of sulprostone only in the stomach. IBMX potentiated the HCO3- response to sulprostone in the duodenum, whereas verapamil reduced the response in both the stomach and duodenum. CONCLUSIONS: PGE stimulates HCO3- secretion via different EP-receptor subtypes in the stomach and duodenum: in the stomach, EP1 receptors are linked to Ca2+; in the duodenum, EP3 receptors are coupled with both adenosine 3', 5'-cyclic monophosphate and Ca2+. PMID- 9352858 TI - Mast cell-dependent tumor necrosis factor alpha production participates in allergic gastric inflammation in mice. AB - BACKGROUND & AIMS: Immunoglobulin E-dependent gastric inflammation is characterized by neutrophil infiltration, and mast cells are required for this response. The aim of this study was to examine whether mast cell production of tumor necrosis factor (TNF)-alpha participates in the recruitment of neutrophils during this response. METHODS: The levels of TNF-alpha messenger RNA (mRNA) and protein in gastric tissues were assessed by Northern blot analysis and enzyme linked immunosorbent assay. In situ hybridization and histochemical staining were performed to identify the cells expressing TNF-alpha transcripts. Anti-TNF-alpha antibodies or cyclosporine A were used in an attempt to inhibit neutrophil infiltration. RESULTS: TNF-alpha mRNA and protein were increased in gastric tissues undergoing immunoglobulin E-dependent inflammation. Mast cells were required for the development of cells expressing TNF-alpha transcripts in the stomach. Seventy-nine percent of the cells in the mucosa and 100% of the cells in the submucosa expressing TNF-alpha mRNA were identified as mast cells. Anti-TNF alpha antibodies inhibited neutrophil infiltration in the submucosa, and cyclosporine A inhibited the tissue expression of TNF-alpha mRNA and the influx of neutrophils into the submucosa and muscularis propria. CONCLUSIONS: These findings show that mast cell-derived TNF-alpha is at least one of the mediators involved in the recruitment of neutrophils during immunoglobulin E-dependent gastric inflammation in the mouse. PMID- 9352859 TI - Water extract of Helicobacter pylori inhibits duodenal mucosal alkaline secretion in anesthetized rats. AB - BACKGROUND & AIMS: The pathophysiology behind Helicobacter pylori-induced gastroduodenal dysfunction is incompletely understood. The aim of this study was to investigate if a water extract of H. pylori distorts acid-induced duodenal mucosal alkaline secretion. METHODS: Chloralose-anesthetized rats were prepared for duodenal luminal perfusion and in situ pH-stat titration of mucosal alkaline secretion. RESULTS: Mucosal bicarbonate secretion increased approximately 55%-60% after a 5-minute exposure to 10 mmol/L HCl. This response was absent when water extracts of three strains of H. pylori (protein content, 0.2-20 microg/mL) had been added to the perfusate. Presence of 3 mmol/L L-arginine, but not the stereoisomer D-arginine, in the luminal perfusate reversed the H. pylori extract blockade of acid-induced mucosal alkaline secretion. High-performance liquid chromatography-based analyses showed that the endogenous nitric oxide synthase inhibitor asymmetric dimethyl arginine (ADMA) increased fourfold in duodenal perfusate and fivefold in duodenal tissue after H. pylori extract exposure. In vitro proteolysis of H. pylori extract also resulted in a substantial accumulation of ADMA. Exogenously administered ADMA, giving similar tissue concentrations, inhibited the mucosal alkaline response to acid exposure. CONCLUSIONS: Water extracts of H. pylori inhibit acid-induced mucosal alkaline secretion via interference with mucosal NO synthase. PMID- 9352860 TI - Glycine-extended gastrin acts as an autocrine growth factor in a nontransformed colon cell line. AB - BACKGROUND & AIMS: The hypothesis that progastrin-derived peptides act as autocrine growth factors for colorectal carcinomas has generated considerable interest. However, the influence of autocrine gastrins on nontumorigenic colonic cells has not been investigated. This study tested the above hypothesis in the nontumorigenic, conditionally immortalized mouse colon cell line YAMC. METHODS: The effects of expression of antisense or sense gastrin messenger RNA, treatment with antibodies against progastrin-derived peptides, or treatment with gastrin receptor antagonists on YAMC cell proliferation were measured. RESULTS: YAMC clones expressing antisense gastrin messenger RNA had reduced levels of immunoreactive progastrin-derived peptides and a reduced rate of proliferation, relative to vector only-transfected cells. Glycine-extended gastrin17, but not amidated gastrin17, reversed the antisense-induced inhibition of proliferation and stimulated the proliferation of sense- or vector only-transfected cells. YAMC cells bound 125I-glycine-extended gastrin17 (Kd, 0.36 nmol/L, 1810 sites/cell), but not 125I-amidated gastrin17, and binding was unaffected by gastrin receptor antagonists including benzotript. Proliferation of all YAMC clones was partially inhibited either by an antibody selective for glycine-extended gastrin or by preincubation with benzotript, and the inhibitory effects were additive. CONCLUSIONS: YAMC cells use nonamidated progastrin-derived peptides as autocrine growth factors, partly through binding to an extracellular receptor selective for glycine-extended gastrin, and partly through an intracellular mechanism. PMID- 9352861 TI - In vivo activation of mitogen-activated protein kinases in rat intestinal neoplasia. AB - BACKGROUND & AIMS: To investigate whether mitogen-activated protein kinase (MAPK) cascades might play a role in the progression of colon cancer, c-Jun N-terminal kinase (JNK) and extracellular signal regulating kinase (ERK) activity during colonic tumorigenesis were examined. METHODS: The 1,2-dimethylhydrazine (DMH) induced colon carcinoma model was used to study the activation of these kinases during intestinal carcinogenesis. Male Sprague-Dawley rats were injected with DMH for 24 weeks. Normal-appearing intestinal mucosa from control and treated animals and DMH-induced intestinal tumors were assayed for JNK and ERK activity using solid phase in vitro kinase assays. Tumors were typed for mutations in the K-ras gene. RESULTS: There was little or no difference in JNK and ERK activity in hyperproliferative mucosa from DMH-treated animals compared with normal mucosa from control animals. However, in 16 colonic neoplasms, an average of 23-fold and 29-fold increases in JNK and ERK activities were observed, respectively, over control levels. In addition, activating protein-1 binding was strongly induced in the colonic tumors. Activation did not correlate with the presence of mutations in K-ras. CONCLUSIONS: Both the JNK and ERK MAPKs are highly activated during late progression of colorectal carcinoma. This change is dependent on the tumorigenic state rather than changes in proliferation. PMID- 9352862 TI - Induction of sodium-dependent bile acid transporter messenger RNA, protein, and activity in rat ileum by cholic acid. AB - BACKGROUND & AIMS: The ileal sodium-dependent bile acid transporter reclaims bile acids from the intestinal lumen to preserve their enterohepatic recirculation. The present studies sought to determine the possible role of enteric bile acids in the molecular regulation of the apical bile acid transporter in rat ileal mucosa. METHODS: Paired rats were fed a control diet or control diet plus cholic acid (1%) or ursodeoxycholic acid (1%) for 10 days. Other paired rats underwent biliary diversion for 72 hours, followed by intraduodenal infusion of taurocholate or fluid/electrolytes. Transporter protein, messenger RNA (mRNA), and activity were determined in the distal 15 cm of ileal mucosa. RESULTS: Transporter protein and mRNA levels in cholic acid-fed rats increased approximately threefold above levels in paired rats fed the control diet (P < 0.02). Similarly, sodium-dependent [3H]taurocholate uptake into membrane vesicles from cholic acid-fed rats increased twofold above uptake into vesicles from control-fed rats because of a twofold increase in maximal transport velocity. In biliary-diverted rats (72-96 hours), transporter protein decreased to 57% +/- 5% of paired controls with intact enterohepatic circulation (P < 0.0001). The intraduodenal infusion of taurocholate (24 hours) in biliary-diverted rats resulted in a time-dependent reinduction of transporter protein expression (3.5 fold). CONCLUSIONS: The expression of the ileal apical bile acid transporter is induced at a pretranslational level by free or taurine-conjugated cholic acid within the small intestine. PMID- 9352863 TI - Noninvasive diagnosis of hepatic fibrosis or cirrhosis. AB - BACKGROUND & AIMS: The evaluation of the degree of hepatic fibrosis is especially important in patients with chronic liver disease. The aim of this study was to study the diagnostic accuracy of noninvasive means. METHODS: Sixty-three clinical, biochemical (prothrombin index, gamma-glutamyl transpeptidase and apolipoprotein A1 levels [PGA score]; and hyaluronate, alpha2-macroglobulin, N terminal peptide of type III procollagen, laminin, and transforming growth factor beta1 levels), Doppler ultrasonic, and endoscopic variables were recorded in 243 patients who were divided into four groups: whole, compensated, alcohol compensated, and viral-compensated liver disease. Diagnostic accuracy was evaluated by discriminant analysis; first globally, then by stepwise analysis. RESULTS: In three groups, hyaluronate and prothrombin index were the best predictive factors (accuracy, > or =85%). Accuracy for the diagnosis of cirrhosis varied from 89.5% to 95% with global discriminant analysis and from 91% to 94% with stepwise analysis according to the group. In the compensated group, hyaluronate concentration of > or =60 microg/L had a sensitivity of 97% and a specificity of 73%. Diagnostic accuracy was 87% globally for extensive fibrosis. Prothrombin index and hyaluronate were two independent variables predictive of the area of fibrosis (r2 = .66). CONCLUSIONS: With the use of a few noninvasive criteria, cirrhosis can be correctly diagnosed in 91%-94% of patients with chronic liver disease. Serum hyaluronate concentration is the most sensitive variable for screening. PMID- 9352864 TI - Growth hormone therapy in patients with cirrhosis: a pilot study of efficacy and safety. AB - BACKGROUND & AIMS: The protein catabolic state of cirrhosis is associated with severe growth hormone (GH) resistance, with low levels of insulin-like growth factor (IGF)-I and its major binding protein (IGFBP)-3. The aim of this study was to conduct a randomized, double-blind, placebo-controlled pilot study of GH therapy in 20 cirrhotic patients to assess the reversibility of GH resistance and subsequent impact on protein economy and safety. METHODS: Patients were treated with GH (0.25 IU/kg body wt) or placebo for 7 days. Serum levels of GH, IGF-I, IGFBP-3, and insulin were measured by radioimmunoassay and 24-hour urinary nitrogen by the Kjeldahl technique. RESULTS: IGF-I levels increased only in the GH-treated group (mean, 69.2 +/- SE 7.0 to 170.6 +/- 48.8 ng/mL; P < 0.05) together with IGFBP-3 (1.65 +/- 0.3 to 2.94 +/- 0.6 mg/L; P < 0.005). Cumulative nitrogen balance similarly improved only in the GH group (2.87-24.16 g; P < 0.05). No significant side effects of GH were observed. CONCLUSIONS: GH therapy can overcome the GH resistance of cirrhosis. The resulting improvement in nitrogen economy and possible influences on clinical outcomes will need to be confirmed in controlled studies of longer duration. PMID- 9352865 TI - Transjugular intrahepatic portosystemic stent shunt versus sclerotherapy plus propranolol for variceal rebleeding. AB - BACKGROUND & AIMS: In patients with cirrhosis of the liver, after the first variceal bleeding episode, transjugular intrahepatic portosystemic stent shunting (TIPS) and endoscopic sclerotherapy plus propranolol (ES) were compared regarding prevention of variceal rebleeding and mortality. METHODS: Eighty-three patients with cirrhosis of the liver were randomized to undergo TIPS (n = 42) or ES (n = 41). RESULTS: Median observation time was in 1.6 years in the TIPS group and 1.45 years in the ES group. Cumulative rates of rebleeding were 23% in the TIPS group and 57% in the ES group (P = 0.0001). Hepatic encephalopathy was observed in 29% of the patients in the TIPS group and in 13% of those in the ES group (P = 0.041). Cumulative rates of survival were 69% in the TIPS group and 67% in the ES group (P = 0.62). Mortality rates in both groups were positively correlated with a higher Child's classification. CONCLUSIONS: Although TIPS significantly reduced the rate of rebleeding, survival rates were not improved. Because TIPS is associated with an increased risk of encephalopathy and high rates of shunt dysfunction, which requires reintervention, the procedure cannot be recommended for elective treatment after the first variceal bleeding episode, but it is an effective therapy in patients in whom endoscopic sclerotherapy fails to control bleeding. PMID- 9352866 TI - Effects of isosorbide-5-mononitrate compared with propranolol on first bleeding and long-term survival in cirrhosis. AB - BACKGROUND & AIMS: Isosorbide-5-mononitrate (Is-5-Mn) exerts beneficial hemodynamic effects in portal hypertension, yet the long-term clinical value is uncertain. The aim of this study was to determine the long-term effects of Is-5 Mn vs. propranolol (Pro) on first bleeding, complications, and death in cirrhosis. METHODS: One hundred eighteen patients included in a previously published randomized trial comparing Is-5-Mn (20 mg three times daily) with Pro were followed up for up to 7 years (range, 2-91 months). Fifty-seven patients received Is-5-Mn and 61 received Pro. RESULTS: Thirty episodes of first upper bleeding occurred; 16 were in the Is-5-Mn group. Actuarial probability of bleeding did not differ between the two groups. Endoscopic variceal red signs were the only independent predictors of early bleeding. Of the 52 patients who died, 28 were in the Is-5-Mn group. The likelihood of death was greater among patients assigned to Is-5-Mn than to Pro, but only in patients older than 50 years (72% vs. 48% at 6 years; P = 0.006). Child-Pugh score, bleeding, age, and assignment to Is-5-Mn were independent predictors of death. The likelihood of death without bleeding was also higher (P = 0.05) in the Is-5-Mn group. CONCLUSIONS: Is-5-Mn is as effective as Pro in preventing early bleeding but is associated with higher long-term mortality. PMID- 9352867 TI - Endoscopic assessment of variceal volume and wall tension in cirrhotic patients: effects of pharmacological therapy. AB - BACKGROUND & AIMS: Variceal rupture is believed to occur when variceal wall tension is excessive. The combined use of endosonography, allowing the objective measurement of variceal radius, and endoscopic measurement of transmural variceal pressure may enable assessment of this important parameter. The aim of this study was to assess the effects on variceal hemodynamics of drugs acting through different mechanisms: decreasing portocollateral blood flow (propranolol) or resistance (isosorbide-5-mononitrate [ISMN]). METHODS: Repeated measurements of variceal radius, volume (by endosonography), and transmural pressure (using endoscopic gauge) were performed in 27 cirrhotic patients at baseline and 40 minutes after double-blind administration of placebo (n = 9), propranolol (n = 9), or ISMN (n = 9). RESULTS: Placebo had no effect. Propranolol significantly reduced variceal volume (-32% +/- 26%; P = 0.01), radius (-12% +/- 9%; P < 0.005), and pressure (-26% +/- 10%; P < 0.0001). The resulting decrease in wall tension (-34% +/- 13%; P < 0.0005) exceeded that in transmural pressure (P < 0.01). ISMN reduced transmural variceal pressure (-26% +/- 21%; P < 0.005), but not radius (-3% +/-14%; NS) and volume (-9% +/- 31%; NS). CONCLUSIONS: The combination of endosonography and endoscopic measurement of transmural variceal pressure allows quantitative estimation of variceal wall tension. Propranolol and ISMN reduce similarly transmural variceal pressure. Propranolol, but not ISMN, reduces variceal volume and radius. Therefore, despite similar decreases in variceal wall tension, propranolol may offer a greater therapeutic effect than ISMN in portal hypertension. PMID- 9352868 TI - Viremia after one month of interferon therapy predicts treatment outcome in patients with chronic hepatitis C. AB - BACKGROUND & AIMS: In chronic hepatitis C, interferon alfa induces sustained remission in less than 30% of treated patients. The aim of this study is to analyze viral status early after initiation interferon therapy as a predictor of treatment outcome. METHODS: One hundred eighty-one patients with chronic hepatitis C who had been treated with interferon alfa for 12 months (median follow-up, 49 months) were studied. Viremia and aminotransferase levels at the first and third months of therapy as well as 10 pretreatment variables were assessed as potential independent predictors of sustained response to treatment. RESULTS: Sustained response occurred in 51 patients (28%). At month 1 of treatment, viral persistence accurately predicted nonresponse (predictive value, 95.3; 95% confidence interval, 86.0-98.8; P < 0.0001). Independent predictors of sustained response were undetectable viremia at the first month of therapy (P < 0.001), undetectable viremia at the third month (P < 0.001), younger age (P = 0.006), nonsporadic infection (P = 0.012), and higher pretreatment aspartate aminotransferase levels (P = 0.032). In patients who cleared HCV RNA at month 1 of therapy, the predicted probability of sustained response averaged 70% for those younger than 30 years and diminished by 10% for each decade of age. CONCLUSIONS: Failure to clear HCV RNA at month 1 of treatment is strongly and independently associated with a very low probability of a sustained response to interferon. PMID- 9352869 TI - Efficacy of a second cycle of interferon therapy in patients with chronic hepatitis C. AB - BACKGROUND & AIMS: Approximately 75%-85% of patients with chronic hepatitis C virus (HCV) infection do not have a sustained response when treated with interferon (IFN). Limited information exists on the efficacy of retreatment with IFN alone in these patients. The aim of this study was to define the efficacy of IFN retreatment in chronic hepatitis C. METHODS: Ninety-two patients with chronic hepatitis C who had shown transient or no response to recombinant IFN-alpha were randomly retreated with different schedules of lymphoblastoid IFN-alpha and followed up for 12 months after therapy to define biochemical and virological response. RESULTS: None of 26 initial nonresponders obtained a sustained response with retreatment, independent of the schedule used. Thirteen of 66 patients (20%; 95% confidence interval [CI], 10.9-31.3) with transient response during the primary cycle developed a sustained biochemical and virological response when retreated, including 3 of 41 (7%; 95% CI, 1.5-9.9) of those receiving the same schedule and 10 of 25 (40%; 95% CI, 21.1-61.3; P < 0.004) of those retreated with a higher dosage and for a longer period. Shorter disease duration (P = 0.02), higher alanine aminotransferase (P = 0.002) and lower gamma-glutamyltransferase levels (P = 0.004), HCV genotype other than HCV-1 (P = 0.03), and a negative serum HCV-RNA test at the end of the primary cycle (P = 0.000) were associated with sustained response. CONCLUSIONS: Patients with chronic hepatitis C who have a relapse after a complete response to a 6-month IFN-alpha treatment should be retreated for 12 months. Nonresponders should not be retreated with IFN alone. PMID- 9352870 TI - Long-term follow-up of patients with chronic hepatitis B treated with interferon alfa. AB - BACKGROUND & AIMS: Therapy with interferon alfa (IFN-alpha) leads to remission of disease in one third of patients with chronic hepatitis B. The aim of this study was to better define the long-term prognosis of this outcome. METHODS: One hundred three patients with chronic hepatitis B who underwent IFN-alpha therapy in three clinical trials between 1984 and 1991 were followed up for serological status, biochemical evidence of liver disease, and liver complications or mortality through 1994. RESULTS: Among 103 patients, 31 (30%) responded to therapy with loss of hepatitis B e antigen and viral DNA from serum. Responders were more likely than nonresponders to be women, black, and to have more severe liver disease including cirrhosis (P < 0.05). Up to 11 years (mean, 6.2 years) after therapy, a higher percentage of responders than nonresponders were still negative for hepatitis B e antigen (94% vs. 40%; P < 0.001) and hepatitis B surface antigen (71% vs 8.3%; P < 0.001). Overall, the rate of liver-related complications and death did not differ by IFN-alpha response, but with adjustment for cirrhosis, nonresponders had higher rates of liver-related complications and mortality (hazard ratio, 13.7; 95% confidence interval, 3.0-63.5). CONCLUSIONS: The response to IFN-alpha therapy in chronic hepatitis B is usually a sustained improvement in disease markers and, when cirrhosis is considered, patient outcome. PMID- 9352871 TI - Transmission of hepatitis B by transplantation of livers from donors positive for antibody to hepatitis B core antigen. The National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database. AB - BACKGROUND & AIMS: Organ donors are a potential source of transmissible disease after transplantation. The aim of this study was to evaluate the risk of acquiring hepatitis B among transplantation recipients of livers from donors without serum hepatitis B surface antigen (HBsAg) but with antibody to hepatitis B core antigen (anti-HBc). METHODS: The transplantation experience of four centers between 1989 and 1994 was reviewed. Recipients of livers from 674 donors were considered informative for hepatitis B virus transmission. RESULTS: Hepatitis B developed in 18 of 23 recipients of livers from anti-HBc-positive donors (78%) compared with only 3 of 651 recipients of anti-HBc-negative donor livers (0.5%) (P < 0.0001). HBsAg persisted in all recipients with donor-related hepatitis B. Liver histology showed chronic hepatitis of moderate severity in 2 of 13 recipients at 1 year and 5 of 8 recipients between 1.6 and 4.5 years from transplantation. Liver transplantation from an anti-HBc-positive donor was associated with decreased 4-year survival (adjusted mortality hazard ratio of 2.4; 95% confidence interval, 1.4-4.0). CONCLUSIONS: De novo posttransplantation hepatitis B infection occurs at a high rate in recipients of donors with anti HBc. Transmission of hepatitis B through transplantation suggests that the virus may persist in the liver despite serological resolution of infection. PMID- 9352872 TI - Genes of the major histocompatibility complex class II influence the outcome of hepatitis C virus infection. AB - BACKGROUND & AIMS: The host's immune response may influence the course of hepatitis C virus (HCV) infection. The aim of this study was to investigate the distribution of HLA class II alleles in white subjects who spontaneously recovered from HCV infection compared with that in patients with persistent infection. METHODS: HLA-DRB1 and -DQB1 typing were performed in 103 consecutive patients with persistent HCV infection (HCV antibody positive, HCV RNA positive) and in 25 subjects with transient HCV infection (HCV antibody positive, persistently negative HCV RNA). RESULTS: No significant differences between subjects with transient or persistent infection were observed for age, sex, source of infection, or HCV serotype. The frequency of DQB1*0301 and DRB1*1101 alleles was higher in patients with transient infection than in those with persistent infection (84% vs. 30.8%, 40% vs. 9.8%; P < 0.01 and P < 0.02, respectively [Bonferroni correction]). DRB1 and DQB1 alleles did not influence viral load as an independent factor. Mean Knodell's scores were lower in patients with DQB1*0301 allele (6.12 +/- 0.4) than in those negative for DQB1*0301 (7.37 +/- 0.3; P < 0.05). CONCLUSIONS: Our results suggest that host- rather than virus related factors are probably involved in the spontaneous clearance of HCV. PMID- 9352873 TI - Hepatoprotective effects of insulin-like growth factor I in rats with carbon tetrachloride-induced cirrhosis. AB - BACKGROUND & AIMS: Bioavailability of insulin-like growth factor (IGF-I) is reduced in liver cirrhosis. The aim of this study was to analyze the effect of IGF-I on liver histopathology and function in experimental cirrhosis. METHODS: Rats received CCl4 inhalations for 11 or 30 weeks (protocols 1 and 2, respectively) and were treated with 2 microg x 100 g body wt(-1) x day(-1) IGF-I (group CI + IGF) or saline (group CI) on weeks 13 and 14 (protocol 1) or on weeks 28-30 (protocol 2). Normal rats were studied in parallel. RESULTS: Serum albumin and total protein levels were reduced in CI but not in CI + IGF rats compared with normal rats. Clotting factors II, VII, and X were significantly greater in CI + IGF than in CI rats. Liver lipid peroxidation products were significantly increased in CI but not in CI + IGF rats, and liver fibrosis was less pronounced in CI + IGF than in CI animals. The activities of antioxidant enzymes and mitochondrial transmembrane potential were reduced compared with normal animals in CI but not in CI + IGF rats. CONCLUSIONS: IGF-I improves liver function and reduces oxidative liver damage and fibrosis in rats with compensated or advanced liver cirrhosis. Improved mitochondrial function could play a role in the hepatoprotective effect of this hormone. PMID- 9352875 TI - Role of reactive oxygen intermediates in interleukin 10 release after cold liver ischemia and reperfusion in mice. AB - BACKGROUND & AIMS: Reactive oxygen intermediates and cytokines are key effectors in reperfusion injury after liver ischemia. We hypothesized that reactive oxygen intermediates act as a signal for the release of tumor necrosis factor (TNF) and interleukin 10 (IL-10) after reperfusion of cold-preserved livers. METHODS: An endotoxin-free isolated perfused mouse liver system was designed. Harvested mouse livers were stored at 4 degrees C for 0-28 hours and reperfused for 90 minutes with a warm oxygenated Hank's balanced salt solution (alone or with additives). Cytokine messenger RNA (mRNA) from whole liver was measured by reverse transcription polymerase chain reaction. Cytokine protein levels and liver injury assessed by alanine aminotransferase levels were evaluated in liver effluent during reperfusion. RESULTS: TNF and IL-10 mRNA and protein concentrations were increased after reperfusion of ischemic livers. N-Acetylcysteine and allopurinol dramatically decreased TNF (-64% and -62%) and IL-10 (-49% and -57%) levels in the effluents, as did an inhibitor of the transcription factor NF-kappaB mobilization (-73% and -76% for TNF and IL-10, respectively). Liver injury was decreased by -40%, -43%, and -54% for the three inhibitors, respectively. CONCLUSIONS: Reactive oxygen intermediates are involved in TNF and IL-10 release after reperfusion of cold-preserved livers. PMID- 9352874 TI - Antiangiogenic agents protect liver sinusoidal lining cells from cold preservation injury in rat liver transplantation. AB - BACKGROUND & AIMS: Low temperature preservation causes unique liver injuries to the sinusoidal lining cells characterized by endothelial cell detachment and rounding and Kupffer cell activation. These changes are similar to those observed during the early stages of angiogenesis. The aim of this study was to investigate if cold preservation injury is caused by the activation of angiogenic mechanisms. METHODS: Livers were obtained from rats pretreated with three well-known antiangiogenic agents (minocycline, interferon alfa-2b, and fumagillin) and were stored for various durations in cold preservation solutions. The effects of the drugs were evaluated by morphometric assessment of endothelial cell injury in H&E, trypan blue, and immunostained (TIE2/Tek) biopsy specimens. Graft functions and survival were evaluated in isolated perfused rat liver and arterialized orthotopic liver transplantation models. RESULTS: Sinusoidal lining cell integrity and viability were significantly improved in animals pretreated with the drugs. Reperfusion injury and survival were also better in pretreated animals. Interferon alfa was the most potent agent, reducing injury even in livers preserved in the current most commonly used solution (University of Wisconsin solution). CONCLUSIONS: Cold preservation injury of liver may be the results of angiogenic mechanisms. This novel observation provides a rationale for improved liver preservation using antiangiogenic agents. PMID- 9352876 TI - Effect of fasting on the uptake of bilirubin and sulfobromophthalein by the isolated perfused rat liver. AB - BACKGROUND & AIMS: Occurrence of hyperbilirubinemia after fasting has been recognized for many years. The pathogenesis of this syndrome is unclear. Although recent studies suggest that increased intestinal deconjugation and reabsorption of bilirubin may play a major role in establishment of hyperbilirubinemia during fasting, other studies have suggested that fasting down-regulates intrinsic hepatocyte transport of bilirubin. The present study was designed to examine this possibility in the isolated perfused rat liver. METHODS: Transport of 3H bilirubin and 35S-sulfobromophthalein (BSP) was examined in isolated perfused livers from 48-hour fasted or control rats using a multiple indicator dilution technique. Data were analyzed to quantify single-pass extraction (model independent analysis) and were also fit by computer to the model of Goresky to quantify unidirectional fluxes. RESULTS: Fasting for 48 hours resulted in an approximately 40% reduction in liver weight but had no effect on model-dependent or model-independent parameters of transport. Despite the fact that the liver was smaller, single-pass extraction of bilirubin and BSP by livers from fasted animals did not differ from control, indicating a greater efficiency at uptake of bilirubin and BSP. CONCLUSIONS: Enhanced enterohepatic circulation of bilirubin, not altered hepatic transport, is a major factor in the pathogenesis of fasting induced hyperbilirubinemia. PMID- 9352877 TI - Cathepsin B contributes to bile salt-induced apoptosis of rat hepatocytes. AB - BACKGROUND & AIMS: Bile salt-induced apoptosis is mediated by a trypsin-like nuclear protease. The aims of this study were to identify this protease and to elucidate its mechanistic role in bile salt-induced hepatocyte apoptosis. METHODS: Rats, isolated rat hepatocytes, and a rat hepatoma cell line stably transfected with a bile salt transporter (McNtcp.24) were used for this study. RESULTS: In the bile duct-ligated rat, a threefold increase in apoptosis and a fourfold increase in trypsin-like nuclear protease activity were observed. The nuclear protease activity was purified from bile duct-ligated rats and identified as cathepsin B. Specific, structurally dissimilar cathepsin B inhibitors blocked glycochenodeoxycholate (GCDC)-induced apoptosis in cultured rat hepatocytes. Furthermore, stable transfection of McNtcp.24 cells with the complementary DNA for cathepsin B in the antisense orientation reduced cathepsin B activity and GCDC-induced apoptosis by >75%. Next, cathepsin B cellular localization during apoptosis was determined by immunoblot analysis of nuclear cell fractions, immunocytochemistry, and by determining the compartmentation of expressed cathepsin B fused to green fluorescent protein. All three approaches showed translocation of cathepsin B from the cytoplasm to the nucleus during GCDC induced apoptosis. CONCLUSIONS: The data suggest that translocation of cathepsin B from the cytoplasm to the nucleus is a mechanism contributing to bile salt induced apoptosis of hepatocytes. PMID- 9352878 TI - The human biliary epithelial cell plasma membrane antigen in primary biliary cirrhosis: pyruvate dehydrogenase X? AB - BACKGROUND & AIMS: Patients with primary biliary cirrhosis (PBC) have autoantibodies that react with components of mitochondrial multienzyme complexes. In addition to binding to mitochondria, patients' autoantibodies to the assumed major autoantigen pyruvate dehydrogenase complex (PDC) dihydrolipoamide acetyltransferase (E2) bind to the plasma membrane of biliary epithelial cells (BECs) specifically in PBC. The aim of this study was to characterize BEC plasma membrane antigens recognized by patients' autoantibodies in PBC. METHODS: Antigens prepared from intracellular and plasma membrane-enriched fractions of BECs purified from PBC and control liver were immunoblotted with anti-PDC. RESULTS: In the intracellular fraction, anti-PDC recognized BEC protein bands corresponding to the molecular weight value of E2 and X components of human heart PDC on Western blots. No difference was observed between PDC-E2 in BECs from PBC and controls. However, in PBC but not controls, a 50-kilodalton antigen was detected in the plasma membrane-enriched fraction. This antigen comigrated with component X of purified human heart PDC and was recognized by antibodies specific for PDC-X. CONCLUSIONS: The data suggest that PDC-X or a cross-reactive 50 kilodalton antigen is the BEC plasma membrane antigen recognized by patients' autoantibodies in PBC. Furthermore, this antigen, rather than PDC-E2, may be a major B-cell target antigen in PBC. PMID- 9352879 TI - Functional expression of the apical Na+-dependent bile acid transporter in large but not small rat cholangiocytes. AB - BACKGROUND & AIMS: Bile acids interact with cholangiocytes, resulting in cholangiocyte proliferation and increases in ductal bile secretion in large but not small cholangiocytes. It was proposed that for bile acids to exert these effects on cholangiocytes, a specific uptake mechanism must be present in cholangiocytes. The aim of this study was to show the expression of a bile acid transporter in cholangiocytes. METHODS: Small and large cholangiocytes or intrahepatic bile duct units (IBDUs) were isolated from normal rats, and gene expression for the apical Na+-dependent bile acid transporter (ABAT) and the 14 kilodalton ileal cytosolic binding protein (IBABP) was assessed by ribonuclease protection assays. Tissue and subcellular distribution of bile acid transporters was also studied. [14C]-Taurocholate uptake into cholangiocytes was determined. RESULTS: Both ABAT and IBABP messenger RNAs were detected in large but not small cholangiocytes. By immunohistochemistry, ABAT was present in large but not small cholangiocytes. Immunofluorescence showed ABAT to be present in the apical membrane of large IBDUs. A Na+-dependent saturable uptake of taurocholate was present in large but not small cholangiocytes. CONCLUSIONS: These proteins may mediate bile acid uptake from the duct lumen in large ducts, resulting in modification of canalicular bile secretion and modulation of ductal bile secretion and growth. PMID- 9352880 TI - Gene targeting demonstrates additive detrimental effects of interleukin 1 and tumor necrosis factor during pancreatitis. AB - BACKGROUND & AIMS: During severe pancreatitis, interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha are produced in large quantities. The aim of this study was to determine whether either one plays a more dominant role and if their detrimental effects are additive. METHODS: Necrotizing pancreatitis was induced in transgenic (-/-) knockout mice deficient in either IL-1 type 1 receptors, TNF type 1 receptors, or both IL-1 and TNF type 1 receptors. Wild-type mice served as controls. Mortality was assessed for 10 days. Additional animals were killed on days 0, 1, 2, 3, and 4 for determination of pancreatitis severity. RESULTS: All three knockout groups showed decreased amylase and lipase, histological score, serum IL-6, and mortality compared with wild-type groups. Animals devoid of receptors for both cytokines showed improved survival and decreased IL-6 levels compared with those devoid of either IL-1 or TNF receptors individually, yet they failed to show a further decrease in pancreatitis severity. CONCLUSIONS: Preventing the activity of IL-1beta or TNF-alpha has a nearly identical beneficial effect on the severity and mortality of acute pancreatitis. Preventing the activity of both cytokines concurrently has no additional effect on pancreatitis severity but further attenuates the systemic stress response and is associated with an additional but modest decrease in mortality. PMID- 9352881 TI - Epidermal growth factor inhibits bombesin-induced activation of phospholipase C beta1 in rat pancreatic acinar cells. AB - BACKGROUND & AIMS: Epidermal growth factor (EGF) inhibits bombesin-induced activation of phosphoinositide-specific phospholipase C (PLC) in pancreatic acini. The aim of this study was to investigate the mechanism by which EGF inhibits bombesin-induced activation of PLC. METHODS: Intact pancreatic acini were pretreated with pertussis toxin to study the role of Gi/o-type heterotrimeric guanosine triphosphate-binding regulatory proteins (G proteins) in EGF-induced modulation of PLC activity. To identify the PLC isoenzyme(s) and Gi/o protein subtype(s) involved in EGF-induced signaling, PLC activity was measured in isolated pancreatic acinar membranes that had been preincubated with immunoneutralizing antibodies raised against various PLC-beta isoenzymes or G protein alpha-subunits. The association of PLC-beta1 and Gi/o-type G proteins was studied by pertussis toxin-catalyzed [32P]adenosine diphosphate ribosylation of PLC-beta1 immunoprecipitates. RESULTS: Pertussis toxin pretreatment of pancreatic acini abolished the inhibitory effect of EGF on bombesin-induced PLC activation and amylase release. Anti-PLC-beta1, -beta3, and Gq/11alpha antibodies inhibited bombesin-induced PLC activity by 50%, 35%, and 65%, respectively. Anti-Gi1 2alpha, but not a Gi3alpha-specific antibody, abolished the inhibitory effect of EGF on bombesin-induced PLC activity. Pertussis toxin-sensitive G proteins coimmunoprecipitated with PLC-beta1 in an EGF-dependent fashion. CONCLUSIONS: EGF inhibits bombesin-induced activation of PLC-beta1 by a mechanism involving activation of Gi1-2 proteins in pancreatic acinar membranes. PMID- 9352882 TI - Cell volume changes modulate cholecystokinin- and carbachol-stimulated amylase release in isolated rat pancreatic acini. AB - BACKGROUND & AIMS: Changes in cell volume have been recently identified as modulators of cell function and gene expression. This study evaluated the regulation of exocrine secretion by pancreatic acini on the basis of changes in cell hydration. METHODS: Acini were exposed to hypotonicity or hypertonicity. The effects of corresponding changes in cell volume on various cell functions were analyzed. RESULTS: Hypertonicity and hypotonicity caused a stepwise cell shrinkage and swelling, respectively. Cell shrinkage decreased and cell swelling increased amylase secretion stimulated by cholecystokinin (CCK) and carbachol but not by secretin. Changes in cell volume did not alter basal or CCK-stimulated calcium concentrations or CCK-stimulated inositol triphosphate generation. The regulation of secretion by cell volume is not mediated via changes in CCK receptor binding or protein kinase C. The increase of amylase release caused by hypotonicity was completely inhibited by cytochalasin B, colchicine, and genistein. Hypotonicity as well as CCK caused activation of mitogen-activated protein kinases. CONCLUSIONS: Changes in cell volume regulate exocrine secretion of pancreatic acini. The effects were found only for secretagogues that act via the calcium/inositol-trisphosphate pathway. However, the mechanisms involved are located at luminal parts of the signal-transduction cascade and involve the cytoskeleton, protein phosphorylation, and activation of mitogen-activated protein kinases. PMID- 9352883 TI - Short-bowel syndrome in children and adults. AB - Short-bowel syndrome is the malabsorptive state that follows extensive resection of the small intestine. Potential long-term survival without parenteral nutrition heavily depends on stimulation of the process of intestinal adaptation, through which the remaining small intestine gradually increases its absorptive capacity. This process is heavily nutrient dependent, and aggressive use of enteral nutrition is required to stimulate its completion. A combination of osmotic sensitivities, nutrient malabsorption, bowel dilatation and dysmotility, and changes in bacterial flora influence the symptoms and the management of this disorder. Chronic complications include parenteral nutrition-induced liver disease, nutrient deficiency states, and, frequently, small bowel bacterial overgrowth. Intestinal transplantation has been successfully developed in some centers in the United States, and preliminary experience suggest a long-term survival of 50%-75%, better in patients receiving an isolated intestinal transplant than a combined liver/bowel transplant. The ultimate role of intestinal transplantation is still undergoing evaluation. PMID- 9352884 TI - Paneth cells and innate immunity in the crypt microenvironment. AB - Paneth cells release granules into the lumen of the crypts of Lieberkuhn in the small intestine where their component proteins participate in mucosal immunity. The granules contain a number of proteins associated with roles in host defense, including lysozyme, secretory phospholipase A2, and alpha-defensins, termed cryptdins. Mouse cryptdins 1-6 and recombinant human Paneth cell alpha-defensin HD-5 are potent antimicrobial agents against certain microorganisms. As defensins, they kill microbes by disruption of the target cell membrane. The peptides are coded by individual, two-exon genes that map to homologous regions of chromosome 8 in mice and humans, and the differential expression of certain mouse cryptdin genes provides markers for studies of crypt ontogeny and epithelial cell differentiation and lineage determination. Neutrophil alpha defensin peptides exhibit numerous biological activities in addition to antimicrobial function including regulation of cell volume, chemotaxis, mitogenicity, and inhibition of natural killer cell activity. When administered apically, mouse cryptdins 2 and 3 can reversibly stimulate human T-84 intestinal epithelial cells to secrete chloride ion, suggesting that alpha-defensins from Paneth cells also may be multifunctional. Thus, cryptdins and varied Paneth cell secretory products seem to contribute both to innate immunity of the crypt lumen and to defining the apical environment of neighboring cells. PMID- 9352885 TI - American Gastroenterological Association. The centennial year: the development of important ideas during the last 100 years. The enigmatic electrical slow wave: a mirror on 100 years of research in gut motility. PMID- 9352886 TI - The generalist-specialist game: the rules are still changing. PMID- 9352887 TI - The pathophysiology of diabetic gastroenteropathy: more vague than vagal. PMID- 9352888 TI - Beta-adrenergic blockers and nitrovasodilators for the treatment of portal hypertension: the good, the bad, the ugly. PMID- 9352889 TI - Fasting hyperbilirubinemia: unraveling the mechanism involved. PMID- 9352890 TI - Dyspepsia algorithms and the value of endoscopy. PMID- 9352891 TI - Teaching old enterocytes new tricks. PMID- 9352892 TI - Confirmation of the Fas counterattack in host-tumor relations. PMID- 9352893 TI - Pathogenesis of MALT lymphoma: an antigen-dependent process. PMID- 9352894 TI - Significance of WBC differential when leukopenia is induced by 6-MP for IBD. PMID- 9352895 TI - Liver cyclooxygenases in alcoholic liver disease. PMID- 9352896 TI - Circling back to gene vaccines. PMID- 9352897 TI - Peritoneal involvement in colonic cancer. PMID- 9352898 TI - Positive regulation of Shigella flexneri virulence genes by integration host factor. AB - In Shigella flexneri, expression of the plasmid-encoded virulence genes is regulated via a complex cascade involving DNA topology, specific transactivators, and the nucleoid-associated protein H-NS, which represses transcription under inappropriate environmental conditions. We have investigated the involvement of a second nucleoid-associated protein, integration host factor (IHF), in virulence gene expression. We found that transcription of the invasion-specific genes is repressed in a strain harboring an ihfA mutation, particularly on entry into the stationary phase. Expression of the virB gene, whose product is required for the activation of these structural genes, is also enhanced by IHF in the stationary phase. In contrast, the virF gene, which encodes an activator of virB, is stimulated by IHF in both the logarithmic and early stationary phases of growth, as is another virF-regulated gene, icsA. We have identified regions of the virF, virB, and icsA promoters which form IHF-dependent protein-DNA complexes in vitro and have located sequences within these regions with similarity to the consensus IHF binding site. Moreover, results from experiments in which the virF or virB gene was expressed constitutively confirm that IHF has a direct input at the level of both virF and virB transcription. Finally, we provide evidence that at the latter promoter, the primary role of IHF may be to overcome repression by the H-NS protein. To our knowledge, this is the first report of a role for IHF in controlling gene expression in S. flexneri. PMID- 9352899 TI - Evolutionary genetics of the isocitrate dehydrogenase gene (icd) in Escherichia coli and Salmonella enterica. AB - Sequences of the icd gene, encoding isocitrate dehydrogenase (IDH), were obtained for 33 strains representing the major phylogenetic lineages of Escherichia coli and Salmonella enterica. Evolutionary relationships of the strains based on variation in icd are generally similar to those previously obtained for several other housekeeping and for invasion genes, but the sequences of S. enterica subspecies V strains are unusual in being almost intermediate between those of the other S. enterica subspecies and E. coli. For S. enterica, the ratio of synonymous (silent) to nonsynonymous (replacement) nucleotide substitutions between pairs of strains was larger than comparable values for 12 other housekeeping and invasion genes, reflecting unusually strong purifying selection against amino acid replacement in the IDH enzyme. All amino acids involved in the catalytic activity and conformational changes of IDH are strictly conserved within and between species. In E. coli, the level of variation at the 3' end of the gene is elevated by the presence in some strains of a 165-bp replacement sequence supplied by the integration of either lambdoid phage 21 or defective prophage element e14. The 72 members of the E. coli Reference Collection (ECOR) and five additional E. coli strains were surveyed for the presence of phage 21 (as prophage) by PCR amplification of a phage 21-specific fragment in and adjacent to the host icd, and the sequence of the phage 21 segment extending from the 3' end of icd through the integrase gene (int) was determined in nine strains of E. coli. Phage 21 was found in 39% of E. coli strains, and its distribution among the ECOR strains is nonrandom. In two ECOR strains, the phage 21 int gene is interrupted by a 1,313-bp insertion element that has 99.3% nucleotide sequence identity with IS3411 of E. coli. The phylogenetic relationships of phage 21 strains derived from sequences of two different genomic regions were strongly incongruent, providing evidence of frequent recombination. PMID- 9352900 TI - Effects of different carbon fluxes on G1 phase duration, cyclin expression, and reserve carbohydrate metabolism in Saccharomyces cerevisiae. AB - By controlled addition of galactose to synchronized galactose-limited Saccharomyces cerevisiae cultures, the growth rate could be regulated while external conditions were kept constant. By using this method, the G1 phase duration was modulated and expression of cell cycle-regulated genes was investigated. The expression of the cyclin genes CLN1 and CLN2 was always induced just before bud emergence, indicating that this event marks the decision to pass Start. Thus, G1 phase elongation was not due to a slower accumulation of the CLN1 and CLN2 mRNA levels. Only small differences in CLN3 expression levels were observed. The maximal SWI4 expression preceded maximal CLN1 and CLN2 expression under all conditions, as expected for a transcriptional activator. But whereas SWI4 was expressed at about 10 to 20 min, before CLN1 and CLN2 expression at high growth rates, this time increased to about 300 min below a particular consumption rate at which the G1 phase strongly elongated. In the slower-growing cultures, also an increase in SWI6 expression was observed in the G1 phase. The increase in G1 phase duration below a particular consumption rate was accompanied by a strong increase in the reserve carbohydrate levels. These carbohydrates were metabolized again before bud emergence, indicating that below this consumption rate, a transient increase in ATP flux is required for progression through the cell cycle. Since Start occurred at different cell sizes under different growth conditions, it is not just a certain cell size that triggers passage through Start. PMID- 9352901 TI - Cloning and expression of a gene cluster encoding three subunits of membrane bound gluconate dehydrogenase from Erwinia cypripedii ATCC 29267 in Escherichia coli. AB - We have cloned the gene cluster encoding three subunits of membrane-bound gluconate dehydrogenase (GADH) from Erwinia cypripedii ATCC 29267 in Escherichia coli by performing a direct-expression assay. The positive clone converted D gluconate to 2-keto-D-gluconate (2KDG) in the culture medium. Nucleotide sequence analysis of the GADH clone revealed that the cloned fragment contained the complete structural genes for a 68-kDa dehydrogenase subunit, a 47-kDa cytochrome c subunit, and a 24-kDa subunit of unknown function and that the genes were clustered with the same transcriptional polarity. Comparison of the deduced amino acid sequences and the NH2-terminal sequences determined for the purified protein indicated that the dehydrogenase, cytochrome c, and 24-kDa subunits contained typical signal peptides of 22, 19, and 42 amino acids, respectively. The molecular masses of the processed subunits deduced from the nucleotide sequences (65, 45, and 20 kDa) coincided well with the molecular masses of subunits estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In E. cypripedii and recombinant E. coli, the GADH was constitutively formed and the activity of GADH was enhanced more than twofold by addition of D-gluconate to the medium. The holoenzyme glucose dehydrogenase of E. coli was reconstituted by addition of pyrroloquinoline quinone to the culture medium, and the conversion of D-glucose or D-gluconate to 2KDG by recombinant E. coli harboring the cloned GADH gene was attempted in batch culture. The conversion yields for D-glucose were 0.95 mol of 2KDG/mol of D-glucose after 16 h of cultivation, and those for D gluconate were 0.95 mol of 2KDG/mol of D-gluconate after 12 h of cultivation. PMID- 9352902 TI - Thermosensing properties of mutant aspartate chemoreceptors with methyl-accepting sites replaced singly or multiply by alanine. AB - The aspartate chemoreceptor Tar has a thermosensing function that is modulated by covalent modification of its four methylation sites (Gln295, Glu302, Gln309, and Glu491). Without posttranslational deamidation, Tar has no thermosensing ability. When Gln295 and Gln309 are deamidated to Glu, the unmethylated and heavily methylated forms function as warm and cold sensors, respectively. In this study, we carried out alanine-scanning mutagenesis of the methylation sites. Although alanine substitutions influenced the signaling bias and the methylation level, all of the mutants retained aspartate-sensing function. Those with single substitutions had almost normal thermosensing properties, indicating that substitutions at any particular methylation site do not seriously impair thermosensing function. In the posttranslational modification-defective background, some of the alanine substitutions restored thermosensing ability. Warm sensors were found among mutants retaining two glutamate residues, and cold sensors were found among those with one or no glutamate residue. This result suggests that the negative charge at the methylation sites is one factor that determines thermosensor phenotypes, although the size and shape of the side chain may also be important. The warm, cold, and null thermosensor phenotypes were clearly differentiated, and no intermediate phenotypes were found. Thus, the different thermosensing phenotypes that result from covalent modification of the methylation sites may reflect distinct structural states. Broader implications for the thermosensing mechanism are also discussed. PMID- 9352904 TI - Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. AB - To map the incidence of natural competence in the genus Streptococcus, we used PCR to screen a number of streptococcal strains for the presence of the recently identified competence regulation operon, containing the comC, -D, and -E genes. This approach established that the operon is present in strains belonging to the S. mitis and S. anginosus groups, but it was not detected in the other strains examined. Competence is induced in S. pneumoniae and S. gordonii by strain specific peptide pheromones, competence-stimulating peptides (CSPs). With its unique primary structure, each CSP represents a separate pheromone type (pherotype), which is recognized by the signalling domain of the downstream histidine kinase, ComD. Thus, all bacteria induced to competence by a particular CSP belong to the same pherotype. In this study, we identified a number of new pherotypes by sequencing the genes encoding the CSP and its receptor from different streptococcal species. We found that in several cases, these genes have a mosaic structure which must have arisen as the result of recombination between two distinct allelic variants. The observed mosaic blocks encompass the region encoding the CSP and the CSP-binding domain of the histidine kinase. Consequently, the recombination events have led to switches in pherotype for the strains involved. This suggests a novel mechanism for the adaptation of naturally competent streptococci to new environmental conditions. PMID- 9352903 TI - Structural and genetic analysis of a mutant of Rhodobacter sphaeroides WS8 deficient in hook length control. AB - Motility in the photosynthetic bacterium Rhodobacter sphaeroides is achieved by the unidirectional rotation of a single subpolar flagellum. In this study, transposon mutagenesis was used to obtain nonmotile flagellar mutants from this bacterium. We report here the isolation and characterization of a mutant that shows a polyhook phenotype. Morphological characterization of the mutant was done by electron microscopy. Polyhooks were obtained by shearing and were used to purify the hook protein monomer (FlgE). The apparent molecular mass of the hook protein was 50 kDa. N-terminal amino acid sequencing and comparisons with the hook proteins of other flagellated bacteria indicated that the Rhodobacter hook protein has consensus sequences common to axial flagellar components. A 25-kb fragment from an R. sphaeroides WS8 cosmid library restored wild-type flagellation and motility to the mutant. Using DNA adjacent to the inserted transposon as a probe, we identified a 4.6-kb SalI restriction fragment that contained the gene responsible for the polyhook phenotype. Nucleotide sequence analysis of this region revealed an open reading frame with a deduced amino acid sequence that was 23.4% identical to that of FliK of Salmonella typhimurium, the polypeptide responsible for hook length control in that enteric bacterium. The relevance of a gene homologous to fliK in the uniflagellated bacterium R. sphaeroides is discussed. PMID- 9352905 TI - CelG from Clostridium cellulolyticum: a multidomain endoglucanase acting efficiently on crystalline cellulose. AB - The gene coding for CelG, a family 9 cellulase from Clostridium cellulolyticum, was cloned and overexpressed in Escherichia coli. Four different forms of the protein were genetically engineered, purified, and studied: CelGL (the entire form of CelG), CelGcat1 (the catalytic domain of CelG alone), CelGcat2 (CelGcat1 plus 91 amino acids at the beginning of the cellulose binding domain [CBD]), and GST-CBD(CelG) (the CBD of CelG fused to glutathione S-transferase). The biochemical properties of CelG were compared with those of CelA, an endoglucanase from C. cellulolyticum which was previously studied. CelG, like CelA, was found to have an endo cutting mode of activity on carboxymethyl cellulose (CMC) but exhibited greater activity on crystalline substrates (bacterial microcrystalline cellulose and Avicel) than CelA. As observed with CelA, the presence of the nonhydrolytic miniscaffolding protein (miniCipC1) enhanced the activity of CelG on phosphoric acid swollen cellulose (PASC), but to a lesser extent. The absence of the CBD led to the complete inactivation of the enzyme. The abilities of CelG and GST-CBD(CelG) to bind various substrates were also studied. Although the entire enzyme is able to bind to crystalline cellulose at a limited number of sites, the chimeric protein GST-CBD(CelG) does not bind to either of the tested substrates (Avicel and PASC). The lack of independence between the two domains and the weak binding to cellulose suggest that this CBD-like domain may play a special role and be either directly or indirectly involved in the catalytic reaction. PMID- 9352907 TI - Molecular evolution and host adaptation of Bordetella spp.: phylogenetic analysis using multilocus enzyme electrophoresis and typing with three insertion sequences. AB - A total of 188 Bordetella strains were characterized by the electrophoretic mobilities of 15 metabolic enzymes and the distribution and variation in positions and copy numbers of three insertion sequences (IS). The presence or absence of IS elements within certain lineages was congruent with estimates of overall genetic relationships as revealed by multilocus enzyme electrophoresis. Bordetella pertussis and ovine B. parapertussis each formed separate clusters, while human B. parapertussis was most closely related to IS1001-containing B. bronchiseptica isolates. The results of the analysis provide support for the hypothesis that the population structure of Bordetella is predominantly clonal, with relatively little effective horizontal gene flow. Only a few examples of putative recombinational exchange of an IS element were detected. Based on the results of this study, we tried to reconstruct the evolutionary history of different host-adapted lineages. PMID- 9352906 TI - Reduction of the periplasmic disulfide bond isomerase, DsbC, occurs by passage of electrons from cytoplasmic thioredoxin. AB - The Escherichia coli periplasmic protein DsbC is active both in vivo and in vitro as a protein disulfide isomerase. For DsbC to attack incorrectly formed disulfide bonds in substrate proteins, its two active-site cysteines should be in the reduced form. Here we present evidence that, in wild-type cells, these two cysteines are reduced. Further, we show that a pathway involving the cytoplasmic proteins thioredoxin reductase and thioredoxin and the cytoplasmic membrane protein DsbD is responsible for the reduction of these cysteines. Thus, reducing potential is passed from cytoplasmic electron donors through the cytoplasmic membrane to DsbC. This pathway does not appear to utilize the cytoplasmic glutathione-glutaredoxin pathway. The redox state of the active-site cysteines of DsbC correlates quite closely with its ability to assist in the folding of proteins with multiple disulfide bonds. Analysis of the activity of mutant forms of DsbC in which either or both of these cysteines have been altered further supports the role of DsbC as a disulfide bond isomerase. PMID- 9352908 TI - Promoter-specific repression of fimB expression by the Escherichia coli nucleoid associated protein H-NS. AB - The H-NS protein is a major component of the Escherichia coli nucleoid. Mutations in hns, the gene encoding H-NS, have pleiotropic effects on the cell altering both the expression of a variety of unlinked genes and the inversion rate of the DNA element containing the fimA promoter. We investigated the interaction between H-NS and fimB, the gene encoding the bidirectional recombinase that catalyzes fimA promoter flipping. In beta-galactosidase assays, we found that fimB expression increased approximately fivefold in an hns2-tetR insertion mutant. In gel mobility shift assays with purified H-NS, we have also shown that H-NS bound directly and cooperatively to the fimB promoter region with greater affinity than for any other known H-NS-regulated gene. Furthermore, this high-affinity interaction resulted in a promoter-specific inhibition of fimB transcription. The addition of purified H-NS to an in vitro transcription system yielded a fivefold or greater reduction in fimB-specific mRNA production. However, the marked increase in cellular FimB levels in the absence of H-NS was not the primary cause of the mutant rapid inversion phenotype. These results are discussed in regard to both H-NS as a transcriptional repressor of fimB expression and its role in regulating type 1 pilus promoter inversion. PMID- 9352909 TI - Characterization of DNA topoisomerase activity in two strains of Mycoplasma fermentans and in Mycoplasma pirum. AB - DNA topoisomerases (topos) are essential enzymes that participate in many cellular processes involving DNA. The presence of the DNA-gyrase genes in various mycoplasmas has been reported elsewhere. However, the characterization of DNA topo activity in mycoplasmas has not been previously undertaken. In this study, we characterized the topo activity in extracts of Mycoplasma fermentans K7 and incognitus and in Mycoplasma pirum, as well as in partially purified extract of M. fermentans K7. The topo activity in these microorganisms had the following properties. (i) The relaxation of supercoiled DNA was ATP dependent. (ii) ATP independent relaxation activity was not detected. (iii) Supercoiling of relaxed topoisomers was not observed. (iv) The relaxation activity was inhibited by DNA gyrase and topo IV antagonists (novobiocin and oxolinic acid) and by eukaryotic topo II (m-AMSA [4'-(9-acridylamino)methanesulfon-m-anisidide]) and topo I antagonists (camptothecin). Other eukaryotic topo II antagonists (teniposide and etoposide) did not affect the topo relaxation activity. (v) Two polypeptides of 66 and 180 kDa were found to be associated with the mycoplasma topo activity. These results suggest that the properties of the topo enzyme in these mycoplasma species resemble those of the bacterial topo IV and the eukaryotic and the bacteriophage T4 topo II. The findings that mycoplasma topo is inhibited by both eukaryotic topo II and topo I antagonists and that m-AMSA and camptothecin inhibited the growth of M. fermentans K7 in culture support our conclusion that these mycoplasma species have topo with unique properties. PMID- 9352910 TI - Propanediol utilization genes (pdu) of Salmonella typhimurium: three genes for the propanediol dehydratase. AB - The propanediol utilization (pdu) operon of Salmonella typhimurium encodes proteins required for the catabolism of propanediol, including a coenzyme B12 dependent propanediol dehydratase. A clone that expresses propanediol dehydratase activity was isolated from a Salmonella genomic library. DNA sequence analysis showed that the clone included part of the pduF gene, the pduABCDE genes, and a long partial open reading frame (ORF1). The clone included 3.9 kbp of pdu DNA which had not been previously sequenced. Complementation and expression studies with subclones constructed via PCR showed that three genes (pduCDE) are necessary and sufficient for propanediol dehydratase activity. The function of ORF1 was not determined. Analyses showed that the S. typhimurium propanediol dehydratase was related to coenzyme B12-dependent glycerol dehydratases from Citrobacter freundii and Klebsiella pneumoniae. Unexpectedly, the S. typhimurium propanediol dehydratase was found to be 98% identical in amino acid sequence to the Klebsiella oxytoca propanediol dehydratase; this is a much higher identity than expected, given the relationship between these organisms. DNA sequence analyses also supported previous studies indicating that the pdu operon was inherited along with the adjacent cobalamin biosynthesis operon by a single horizontal gene transfer. PMID- 9352911 TI - Cellular levels of factor 390 and methanogenic enzymes during growth of Methanobacterium thermoautotrophicum deltaH. AB - Methanobacterium thermoautotrophicum deltaH was grown in a fed-batch fermentor and in a chemostat under a variety of 80% hydrogen-20% CO2 gassing regimes. During growth or after the establishment of steady-state conditions, the cells were analyzed for the content of adenylylated coenzyme F420 (factor F390-A) and other methanogenic cofactors. In addition, cells collected from the chemostat were measured for methyl coenzyme M reductase isoenzyme (MCR I and MCR II) content as well as for specific activities of coenzyme F420-dependent and H2 dependent methylenetetrahydromethanopterin dehydrogenase (F420-MDH and H2-MDH, respectively), total (viologen-reducing) and coenzyme F420-reducing hydrogenase (FRH), factor F390 synthetase, and factor F390 hydrolase. The experiments were performed to investigate how the intracellular F390 concentrations changed with the growth conditions used and how the variations were related to changes in levels of enzymes that are known to be differentially expressed. The levels of factor F390 varied in a way that is consistently understood from the biochemical mechanisms underlying its synthesis and degradation. Moreover, a remarkable correlation was observed between expression levels of MCR I and II, F420-MDH, and H2-MDH and the cellular contents of the factor. These results suggest that factor F390 is a reporter compound for hydrogen limitation and may act as a response regulator of methanogenic metabolism. PMID- 9352912 TI - Deletion of the N-terminal region of the AREA protein is correlated with a derepressed phenotype with respect to nitrogen metabolite repression. AB - The entire areA gene and a truncated version lacking the sequence encoding the N terminal 389 amino acids were expressed from the qutE promoter and terminator in an Aspergillus nidulans strain with the endogenous areA gene deleted. This expression system was used to decouple the effects of transcription regulation and mRNA stability mediated by the native promoter and terminator from any posttranslational modulation of AREA activity. Both the full-length AREA protein and the truncated form were able to function in the deletion strain, conferring the ability to use alternate nitrogen sources. Transformants containing the entire areA gene had a repressible phenotype with respect to nitrogen metabolite repression, whereas those containing the truncated form of the areA gene had a derepressed phenotype. The truncated areA gene was expressed in an A. nidulans strain containing a normally regulated wild-type areA gene, and transformants displayed a quinate-inducible nitrogen metabolite derepressed phenotype. Northern blot analysis of transformed strains showed that areA-specific mRNAs of the expected sizes were being produced. The truncated AREA protein was overproduced in Escherichia coli as a fusion protein and purified to homogeneity by a single step immobilized metal affinity chromatography, and the purified protein was shown to bind specifically to the niaD promoter. Revised sequences of the 5' region of the areA gene and the entire meaB gene are reported. PMID- 9352913 TI - Catabolite repression in Lactobacillus casei ATCC 393 is mediated by CcpA. AB - The chromosomal ccpA gene from Lactobacillus casei ATCC 393 has been cloned and sequenced. It encodes the CcpA protein, a central catabolite regulator belonging to the LacI-GalR family of bacterial repressors, and shows 54% identity with CcpA proteins from Bacillus subtilis and Bacillus megaterium. The L. casei ccpA gene was able to complement a B. subtilis ccpA mutant. An L. casei ccpA mutant showed increased doubling times and a relief of the catabolite repression of some enzymatic activities, such as N-acetylglucosaminidase and phospho-beta galactosidase. Detailed analysis of CcpA activity was performed by using the promoter region of the L. casei chromosomal lacTEGF operon which is subject to catabolite repression and contains a catabolite responsive element (cre) consensus sequence. Deletion of this cre site or the presence of the ccpA mutation abolished the catabolite repression of a lacp::gusA fusion. These data support the role of CcpA as a common regulatory element mediating catabolite repression in low-GC-content gram-positive bacteria. PMID- 9352914 TI - Regulation of upp expression in Escherichia coli by UTP-sensitive selection of transcriptional start sites coupled with UTP-dependent reiterative transcription. AB - Expression of the upp gene of Escherichia coli, which encodes the pyrimidine salvage enzyme uracil phosphoribosyltransferase, is negatively regulated by pyrimidine availability. In this study, we demonstrate that this regulation occurs mainly by UTP-sensitive selection of alternative transcriptional start sites, which produces transcripts that differ in the ability to be productively elongated. The upp initially transcribed region contains the sequence GATTTTTTTTG (nontemplate strand). Transcription is initiated primarily at the first two bases in this sequence, designated G6 and A7 (counting from the promoter -10 region). High intracellular levels of UTP favor initiation at position A7; however, the resulting transcripts are subject to reiterative transcription (i.e., repetitive nucleotide addition) within the run of T residues in the initially transcribed region. The resulting AUUUUn (where n = 1 to >50) transcripts are not extended to include downstream upp sequences. In contrast, low intracellular levels of UTP strongly favor initiation at position G6, which results in transcripts that generally do not engage in reiterative transcription and thus can be normally elongated. This mechanism ensures that high levels of uracil phosphoribosyltransferase are produced only under conditions of pyrimidine limitation. The mechanisms that account for UTP-sensitive start site selection and different fates of upp transcripts, as well as the general use of UTP dependent reiterative transcription in gene regulation, are discussed in detail. PMID- 9352915 TI - Involvement of two alpha-ketoglutarate-dependent dioxygenases in enantioselective degradation of (R)- and (S)-mecoprop by Sphingomonas herbicidovorans MH. AB - Cell extracts of Sphingomonas herbicidovorans MH grown on (R)-mecoprop contained an enzyme activity that selectively converted (R)-mecoprop to 4-chloro-2 methylphenol, whereas extracts of cells grown on (S)-mecoprop contained an enzyme activity selective for the S enantiomer. Both reactions were dependent on alpha ketoglutarate and ferrous ions. Besides 4-chloro-2-methylphenol, pyruvate and succinate were detected as products of the reactions. Labeling experiments with (18)O2 revealed that both enzyme activities catalyzed a dioxygenation reaction. One of the oxygen atoms of pyruvate and one of the oxygen atoms of succinate were derived from molecular oxygen. Analysis of cell extracts obtained from cells grown on different substrates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that growth on (R)-mecoprop and (S)-mecoprop caused the appearance of prominent protein bands at 34 and 32 kDa, respectively. Both protein bands were present when cells grew on the racemic mixture. The results demonstrate that S. herbicidovorans initiated the degradation of each enantiomer of mecoprop by a specific alpha-ketoglutarate-dependent dioxygenase. By comparing conversion rates of various phenoxy herbicides, we confirmed that the two enzyme activities were distinct from that of TfdA, which catalyzes the first step in the degradation of 2,4-dichlorophenoxyacetic acid in Ralstonia eutropha JMP134. PMID- 9352916 TI - Purification and properties of 4-hydroxybenzoate 1-hydroxylase (decarboxylating), a novel flavin adenine dinucleotide-dependent monooxygenase from Candida parapsilosis CBS604. AB - A novel flavoprotein monooxygenase, 4-hydroxybenzoate 1-hydroxylase (decarboxylating), from Candida parapsilosis CBS604 was purified to apparent homogeneity. The enzyme is induced when the yeast is grown on either 4 hydroxybenzoate, 2,4-dihydroxybenzoate, or 3,4-dihydroxybenzoate as the sole carbon source. The purified monooxygenase is a monomer of about 50 kDa containing flavin adenine dinucleotide as weakly bound cofactor. 4-Hydroxybenzoate 1 hydroxylase from C. parapsilosis catalyzes the oxidative decarboxylation of a wide range of 4-hydroxybenzoate derivatives with the stoichiometric consumption of NAD(P)H and oxygen. Optimal catalysis is reached at pH 8, with NADH being the preferred electron donor. By using (18)O2, it was confirmed that the oxygen atom inserted into the product 1,4-dihydroxybenzene is derived from molecular oxygen. 19F nuclear magnetic resonance spectroscopy revealed that the enzyme catalyzes the conversion of fluorinated 4-hydroxybenzoates to the corresponding hydroquinones. The activity of the enzyme is strongly inhibited by 3,5-dichloro-4 hydroxybenzoate, 4-hydroxy-3,5-dinitrobenzoate, and 4-hydroxyisophthalate, which are competitors with the aromatic substrate. The same type of inhibition is exhibited by chloride ions. Molecular orbital calculations show that upon deprotonation of the 4-hydroxy group, nucleophilic reactivity is located in all substrates at the C-1 position. This, and the fact that the enzyme is highly active with tetrafluoro-4-hydroxybenzoate and 4-hydroxy-3-nitrobenzoate, suggests that the phenolate forms of the substrates play an important role in catalysis. Based on the substrate specificity, a mechanism is proposed for the flavin mediated oxidative decarboxylation of 4-hydroxybenzoate. PMID- 9352917 TI - Carbon starvation of Salmonella typhimurium does not cause a general increase of mutation rates. AB - Mutation rates in bacteria can vary depending on the genetic target studied and the specific growth conditions of the cells. Here, two different methods were used to determine how rates of mutation to antibiotic resistance, auxotrophy, and prototrophy were influenced by carbon starvation on agar plates. The rate of mutation to rifampin resistance was increased by starvation as measured by fluctuation tests, similar to what has been reported previously for Escherichia coli. In contrast, the rates of mutation to various types of auxotrophy were unaffected or decreased as measured by both fluctuation tests and a repeated streaking procedure. Similarly, the rates of reversion to prototrophy of his and lac nonsense and missense mutations were unaffected by starvation. Thus, mutation rates of different genetic targets can be affected differently by starvation and we conclude that carbon starvation is not generally mutagenic in Salmonella typhimurium. PMID- 9352918 TI - Topological analysis of the membrane-bound glucosyltransferase, MdoH, required for osmoregulated periplasmic glucan synthesis in Escherichia coli. AB - The MdoH protein is essential for synthesis of the osmoregulated periplasmic glucans, known as membrane-derived oligosaccharides (MDOs), in Escherichia coli. Mutants lacking MdoH are deficient in a glucosyltransferase activity assayed in vitro. The MdoH protein is the product of the second gene of an operon, and it has been shown to span the cytoplasmic membrane. The MdoH protein comprises 847 amino acids and is poorly expressed as observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. We have experimentally measured the topological organization of MdoH within the membrane by construction of fusions to beta-lactamase as a reporter. Analysis of 51 different MdoH-beta-lactamase fusions suggested that the MdoH protein crosses the cytoplasmic membrane eight times, with the N and C termini in the cytoplasm. Moreover, a 310-amino-acid domain is present in the cytoplasm between the second and third transmembrane segments. It was deduced from the measurement of the MDO biosynthetic activity of truncated or fused MdoH proteins that almost all the C-terminal residues are necessary for this activity. The model of the MdoH protein in the membrane suggests that this protein could be directly involved in the translocation of nascent polyglucose chains to the periplasmic space. PMID- 9352919 TI - Mutants of Ralstonia (Pseudomonas) solanacearum sensitive to antimicrobial peptides are altered in their lipopolysaccharide structure and are avirulent in tobacco. AB - Ralstonia solanacearum K60 was mutagenized with the transposon Tn5, and two mutants, M2 and M88, were isolated. Both mutants were selected based on their increased sensitivity to thionins, and they had the Tn5 insertion in the same gene, 34 bp apart. Sequence analysis of the interrupted gene showed clear homology with the rfaF gene from Escherichia coli and Salmonella typhimurium (66% similarity), which encodes a heptosyltransferase involved in the synthesis of the lipopolysaccharide (LPS) core. Mutants M2 and M88 had an altered LPS electrophoretic pattern, consistent with synthesis of incomplete LPS cores. For these reasons, the R. solanacearum gene was designated rfaF. The mutants were also sensitive to purified lipid transfer proteins (LTPs) and to an LTP-enriched, cell wall extract from tobacco leaves. Mutants M2 and M88 died rapidly in planta and failed to produce necrosis when infiltrated in tobacco leaves or to cause wilting when injected in tobacco stems. Complemented strains M2* and M88* were respectively obtained from mutants M2 and M88 by transformation with a DNA fragment harboring gene rfaF. They had a different degree of wild-type reconstituted phenotype. Both strains retained the rough phenotype of the mutants, and their LPS electrophoretic patterns were intermediate between those of the wild type and those of the mutants. PMID- 9352920 TI - IF3-mediated suppression of a GUA initiation codon mutation in the recJ gene of Escherichia coli. AB - A mutational change of the initiation codon to GUA was found to reduce, but not abolish, expression of the recJ gene of Escherichia coli. Specific mutations in translational initiation factor IF3 have been isolated as second-site suppressors of this GUA initiation codon mutation. One of these, infC135, with an arginine-to proline change at amino acid 131, completely restores a wild-type phenotype to recJ GUA initiation codon mutants and acts in a semidominant fashion. The infC135 mutation increased expression of RecJ from the GUA mutant but had no effect on the normal GUG start. The infC135 mutation also abolished autoregulation of IF3 in cis and in trans. The behavior of this IF3 mutant suggests that it has specifically lost its ability to abort initiation from poor initiation codons such as GUA of recJ and the AUU of infC. Because of the impact of IF3 on recJ, a recombination and repair gene, this role of IF3 must be general and not restricted to translation genes. The dominance of infC135 suggests that the other functions of IF3, for instance its ability to bind to 30S ribosomes, must remain intact. Although the ability to discriminate among initiation codons has been lost in the infC135 mutant, translational initiation was still restricted to the normal initiation site in recJ, even in the presence of a closely juxtaposed alternative initiation codon. Because the recJ gene lacks a canonical Shine Dalgarno sequence, other unknown features of the mRNA must serve to specify the initiation site. PMID- 9352921 TI - High- and low-abundance chemoreceptors in Escherichia coli: differential activities associated with closely related cytoplasmic domains. AB - In Escherichia coli, two high-abundance chemoreceptors are present in cellular dosages approximately ten-fold greater than two low-abundance receptors. In the absence of high-abundance receptors, cells exhibit an abnormally low tumble frequency and the ability of the remaining receptors to mediate directed migration in spatial gradients is substantially compromised. We found that increasing the cellular amount of the low-abundance receptor Trg over a range of dosages did not alleviate these defects and thus concluded that high- and low abundance receptors are distinguished not simply by their different dosages in a wild-type cell but also by an inherent difference in activity. By creating hybrids of the low-abundance receptor Trg and the high-abundance receptor Tsr, we investigated the possibility that this inherent difference could be localized to a specific receptor domain and found that the cytoplasmic domain of the high abundance receptor Tsr conferred the essential features of that receptor class on the low-abundance receptor Trg, even though it is in this domain that residue identity between the two receptors is substantially conserved. PMID- 9352922 TI - Localization of the active site of the alpha subunit of the Escherichia coli DNA polymerase III holoenzyme. AB - Using a deletion approach on the alpha subunit of DNA polymerase III from Escherichia coli, we show that there is an N-proximal polymerase domain which is distinct from a more C-proximal tau and beta binding domain. Although deletion of 60 residues from the alpha N terminus abolishes polymerase activity, deletions of 48, 169, and 342 amino acids from the C terminus progressively impair its catalytic efficiency but preserve an active site. Deletion of 342 C-terminal residues reduces k(cat) 46-fold, increases the Km for gapped DNA 5.5-fold, and increases the Km for deoxynucleoside triphosphates (dNTPs) twofold. The 818 residue protein with polymerase activity displays typical Michaelis-Menten behavior, catalyzing a polymerase reaction that is saturable with substrate and linear with time. With the aid of newly acquired sequences of the polymerase III alpha subunit from a variety of organisms, candidates for two key aspartate residues in the active site are identified at amino acids 401 and 403 of the E. coli sequence by inspection of conserved acidic amino acids. The motif Pro-Asp-X Asp, where X is a hydrophobic amino acid, is shown to be conserved among all known DnaE proteins, including those from Bacillaceae, cyanobacteria, Mycoplasma, and mycobacteria. The E. coli DnaE deletion protein with only the N-terminal 366 amino acids does not have polymerase activity, consistent with the proposed position of the active-site residues. PMID- 9352923 TI - A tricarboxylic acid cycle intermediate regulating transcription of a chloroaromatic biodegradative pathway: fumarate-mediated repression of the clcABD operon. AB - The ortho-cleavage pathways of catechol and 3-chlorocatechol are central catabolic pathways of Pseudomonas putida that convert aromatic and chloroaromatic compounds to tricarboxylic acid (TCA) cycle intermediates. They are encoded by the evolutionarily related catBCA and clcABD operons, respectively. Expression of the cat and clc operons requires the LysR-type transcriptional activators CatR and ClcR, respectively, and the inducer molecules cis,cis-muconate and 2-chloro cis,cis-muconate, respectively. The regulation of the cat and clc promoters has been well studied, but the extent to which these operons are repressed by growth in TCA cycle intermediates has not been explored. We demonstrate by transcriptional fusion studies that the expression from the clc promoter is repressed when the cells are grown on succinate, citrate, or fumarate and that this repression is ClcR dependent and occurs at the transcriptional level. The presence of these organic acids did not affect the expression from the cat promoter. In vitro transcription assays demonstrate that the TCA cycle intermediate fumarate directly and specifically inhibits the formation of the clcA transcript. No such inhibition was observed when CatR was used as the activator on either the cat or clc template. Titration studies of fumarate and 2 chloromuconate show that the fumarate effect is concentration dependent and reversible, indicating that fumarate and 2-chloromuconate most probably compete for the same binding site on ClcR. This is an interesting example of the transcriptional regulation of a biodegradative pathway by the intracellular sensing of the state of the TCA cycle. PMID- 9352924 TI - Hydrogen sulfide production and fermentative gas production by Salmonella typhimurium require F0F1 ATP synthase activity. AB - A previously isolated mutant of Salmonella typhimurium lacking hydrogen sulfide production from both thiosulfate and sulfite was shown to have a single mutation which also caused the loss of fermentative gas production and the ability to grow on nonfermentable substrates and which mapped in the vicinity of the atp chromosomal locus. The implication that F0F1 ATP synthase might be essential for H2S and fermentative gas production was explored. The phs plasmid conferring H2S production on wild-type Escherichia coli failed to confer this ability on seven of eight E. coli atp point mutants representing, collectively, the eight genes encoding the subunits of F0F1 ATP synthase. However, it did confer some thiosulfate reductase activity on all except the mutant with a lesion in the ATP synthase catalytic subunit. Localized mutagenesis of the Salmonella atp chromosomal region yielded 500 point mutants unable to reduce thiosulfate to H2S or to produce gas from glucose, but differing in the extents of their ability to grow on succinate, to perform proton translocation as measured in a fluorescence quenching assay, and to reduce sulfite to H2S. Biochemical assays showed that all mutants were completely devoid of both methyl viologen and formate-linked thiosulfate reductase and that N,N'-dicyclohexylcarbodiimide blocked thiosulfate reductase activity by the wild type, suggesting that thiosulfate reductase activity has an absolute requirement for F0F1 ATP synthase. Hydrogenase-linked formate dehydrogenase was also affected, but not as severely as thiosulfate reductase. These results imply that in addition to linking oxidation with phosphorylation, F0F1 ATP synthase plays a key role in the proton movement accompanying certain anaerobic reductions and oxidations. PMID- 9352925 TI - A triggered-suicide system designed as a defense against bacteriophages. AB - A novel bacteriophage protection system for Lactococcus lactis based on a genetic trap, in which a strictly phage-inducible promoter isolated from the lytic phage phi31 is used to activate a bacterial suicide system after infection, was developed. The lethal gene of the suicide system consists of the three-gene restriction cassette LlaIR+, which is lethal across a wide range of gram-positive bacteria. The phage-inducible trigger promoter (phi31P) and the LlaIR+ restriction cassette were cloned in Escherichia coli on a high-copy-number replicon to generate pTRK414H. Restriction activity was not apparent in E. coli or L. lactis prior to phage infection. In phage challenges of L. lactis(pTRK414H) with phi31, the efficiency of plaquing was lowered to 10(-4) and accompanied by a fourfold reduction in burst size. Center-of-infection assays revealed that only 15% of infected cells released progeny phage. In addition to phage phi31, the phi31P/LlaIR+ suicide cassette also inhibited four phi31-derived recombinant phages at levels at least 10-fold greater than that of phi31. The phi31P/LlaIR+ based suicide system is a genetically engineered form of abortive infection that traps and eliminates phages potentially evolving in fermentation environments by destroying the phage genome and killing the propagation host. This type of phage triggered suicide system could be designed for any bacterium-phage combination, given a universal lethal gene and an inducible promoter which is triggered by the infecting bacteriophage. PMID- 9352926 TI - Characterization of anaerobic fermentative growth of Bacillus subtilis: identification of fermentation end products and genes required for growth. AB - Bacillus subtilis can grow anaerobically by respiration with nitrate as a terminal electron acceptor. In the absence of external electron acceptors, it grows by fermentation. Identification of fermentation products by using in vivo nuclear magnetic resonance scans of whole cultures indicated that B. subtilis grows by mixed acid-butanediol fermentation but that no formate is produced. An ace mutant that lacks pyruvate dehydrogenase (PDH) activity was unable to grow anaerobically and produced hardly any fermentation product. These results suggest that PDH is involved in most or all acetyl coenzyme A production in B. subtilis under anaerobic conditions, unlike Escherichia coli, which uses pyruvate formate lyase. Nitrate respiration was previously shown to require the ResDE two component signal transduction system and an anaerobic gene regulator, FNR. Also required are respiratory nitrate reductase, encoded by the narGHJI operon, and moaA, involved in biosynthesis of a molybdopterin cofactor of nitrate reductase. The resD and resDE mutations were shown to moderately affect fermentation, but nitrate reductase activity and fnr are dispensable for fermentative growth. A search for genes involved in fermentation indicated that ftsH is required, and is also needed to a lesser extent for nitrate respiration. These results show that nitrate respiration and fermentation of B. subtilis are governed by divergent regulatory pathways. PMID- 9352927 TI - Cloning of the Staphylococcus aureus ddh gene encoding NAD+-dependent D-lactate dehydrogenase and insertional inactivation in a glycopeptide-resistant isolate. AB - The mechanism of low-level glycopeptide resistance among staphylococci is not known. A cytoplasmic protein, provisionally called Ddh (W. M. Milewski, S. Boyle Vavra, B. Moreira, C. C. Ebert, and R. S. Daum, Antimicrob. Agents Chemother. 40:166-172, 1996), and the RNA transcript that contains the ddh gene, which encodes Ddh, are present in increased amounts in a vancomycin-resistant isolate, 523k, compared with the susceptible parent isolate, 523. Sequence analysis had previously revealed that Ddh is related to NAD+-dependent D-lactate dehydrogenase (D-nLDH) and VanH. This latter protein is essential for high-level glycopeptide resistance in Enterococcus faecium and Enterococcus faecalis by synthesizing the D-lactate needed for biosynthesis of D-lactate-terminating peptidoglycan precursors with low affinity for vancomycin. We now provide the direct evidence that the ddh gene product is Staphylococcus aureus D-nLDH and hereafter refer to the protein as D-nLDH. However, overproduction of this protein in isolate 523k did not result in production of D-lactate-containing peptidoglycan precursors, and susceptibility testing of ddh mutants of 523k demonstrated that S. aureus D nLDH is not necessary for glycopeptide resistance in this isolate. We conclude that the mechanism of glycopeptide resistance in this isolate is distinct from that in enterococci. PMID- 9352928 TI - Photoresponses of the purple nonsulfur bacteria Rhodospirillum centenum and Rhodobacter sphaeroides. AB - We have measured the photoresponse of two purple nonsulfur bacteria, Rhodobacter sphaeroides and Rhodospirillum centenum, under defined conditions in a light beam propagating at 90 degrees to the optical axis of the microscope. This beam presented cells with a steep gradient of intensity perpendicular to the direction of propagation and a shallow gradient in the direction of light propagation. R. centenum, a species that reverses to change direction, accumulated in the light beam, as expected for a "scotophobic" response, while R. sphaeroides, which stops rather than reverses, accumulated outside the light beam. We also compared the behavior of liquid-grown R. centenum, which swims by using a single polar flagellum, to that of surface-grown R. centenum, which swarms over agar by using many lateral flagella and has been shown to move as colonies toward specific wavelengths of light. When suspended in liquid medium, both liquid- and surface grown R. centenum showed similar responses to the light gradient. In all cases, free-swimming cells responded to the steep gradient of intensity but not to the shallow gradient, indicating they cannot sense the direction of light propagation but only its intensity. In a control experiment, the known phototactic alga Chlamydamonas reinhardtii was shown to swim in the direction of light propagation. PMID- 9352929 TI - Two isofunctional nitric oxide reductases in Alcaligenes eutrophus H16. AB - Two genes, norB and norZ, encoding two independent nitric oxide reductases have been identified in Alcaligenes eutrophus H16. norB and norZ predict polypeptides of 84.5 kDa with amino acid sequence identity of 90%. While norB resides on the megaplasmid pHG1, the norZ gene is located on a chromosomal DNA fragment. Amino acid sequence analysis suggests that norB and norZ encode integral membrane proteins composed of 14 membrane-spanning helices. The region encompassing helices 3 to 14 shows similarity to the NorB subunit of common bacterial nitric oxide reductases, including the positions of six strictly conserved histidine residues. Unlike the Nor enzymes characterized so far from denitrifying bacteria, NorB and NorZ of A. eutrophus contain an amino-terminal extension which may form two additional helices connected by a hydrophilic loop of 203 amino acids. The presence of a NorB/NorZ-like protein was predicted from the genome sequence of the cyanobacterium Synechocystis sp. strain PCC6803. While the common NorB of denitrifying bacteria is associated with a second cytochrome c subunit, encoded by the neighboring gene norC, the nor loci of A. eutrophus and Synechocystis lack adjacent norC homologs. The physiological roles of norB and norZ in A. eutrophus were investigated with mutants disrupted in the two genes. Mutants bearing single site deletions in norB or norZ were affected neither in aerobic nor in anaerobic growth with nitrate or nitrite as the terminal electron acceptor. Inactivation of both norB and norZ was lethal to the cells under anaerobic growth conditions. Anaerobic growth was restored in the double mutant by introducing either norB or norZ on a broad-host-range plasmid. These results show that the norB and norZ gene products are isofunctional and instrumental in denitrification. PMID- 9352930 TI - Regulation of Bacillus subtilis sigmaH (spo0H) and AbrB in response to changes in external pH. AB - The RNA polymerase sigma subunit, sigmaH, of Bacillus subtilis is required for the transcription of genes that are induced in late-growth cultures at high cell density, including genes that function in sporulation. The expression of sigmaH controlled genes is repressed when nutrient broth sporulation medium (Difco sporulation medium [DSM]) is supplemented with high concentrations of glucose and glutamine (DSM-GG), preferred carbon and nitrogen sources of B. subtilis. Under these conditions, the pH of the DSM-GG medium decreases to approximately 5. Raising the pH by the addition of morpholinepropanesulfonic acid (MOPS) or Tris HCl (pH 7.5) results in a dramatic increase in the expression of lacZ fusions to sigmaH-dependent promoters. Correspondingly, the level of sigmaH protein was higher in cells of late-growth DSM-GG cultures treated with a pH stabilizer. When sigmaH-dependent gene expression was examined in cells bearing a mutation in abrB, encoding the transition state regulator that negatively controls genes transcribed by the sigmaH form of RNA polymerase, derepression was observed as well as an increase in medium pH. Reducing the pH with acetic acid resulted in repression, suggesting that AbrB was not functioning directly in pH-dependent repression but was required to maintain the low medium pH in DSM-GG. AbrB protein levels were high in late-growth, DSM-GG cultures but significantly lower when the pH was raised by Tris-HCl addition. An active tricarboxylic acid (TCA) cycle was required to obtain maximum derepression of sigmaH-dependent transcription, and transcription of the TCA cycle enzyme gene citB was repressed in DSM-GG but derepressed when the pH was artificially raised. The negative effect of low pH on sigmaH-dependent lacZ expression was also observed in unbuffered minimal medium and appeared to be exerted posttranslationally with respect to spo0H expression. However, the addition of amino acids to the medium caused pH-independent repression of both sigmaH-dependent transcription and spo0H-lacZ expression. These results suggest that spo0H transcription or translation is repressed by a mechanism responding to the availability of amino acids whereas spo0H is posttranslationally regulated in response to external pH. PMID- 9352931 TI - Interactions between heterologous FtsA and FtsZ proteins at the FtsZ ring. AB - FtsZ and FtsA are essential for cell division in Escherichia coli and colocalize to the septal ring. One approach to determine what regions of FtsA and FtsZ are important for their interaction is to identify in vivo interactions between FtsA and FtsZ from different species. As a first step, the ftsA genes of Rhizobium meliloti and Agrobacterium tumefaciens were isolated and characterized. In addition, an FtsZ homolog that shared the unusual C-terminal extension of R. meliloti FtsZ1 was found in A. tumefaciens. In order to visualize their localization in cells, we tagged these proteins with green fluorescent protein (GFP). When R. meliloti FtsZ1-GFP or A. tumefaciens FtsZ-GFP was expressed at low levels in E. coli, they specifically localized only to the E. coli FtsZ ring, possibly by coassembly. When A. tumefaciens FtsA-GFP or R. meliloti FtsA-GFP was expressed in E. coli, they failed to localize detectably to the E. coli FtsZ ring. However, when R. meliloti FtsZ1 was coexpressed with them, fluorescence localized to a band at the midcell division site, strongly suggesting that FtsA from either A. tumefaciens or R. meliloti can bind directly to its cognate FtsZ. As expected, GFP-tagged FtsZ1 and FtsA from either R. meliloti or A. tumefaciens localized to the division site in A. tumefaciens cells. Therefore, the 61 amino acid changes between A. tumefaciens FtsA and R. meliloti FtsA do not prevent their direct interaction with FtsZ1 from either species, suggesting that those residues are not essential for protein-protein contacts. Moreover, the failure of the two non-E. coli FtsA derivatives to interact strongly with E. coli FtsZ in this in vivo system unless their cognate FtsZ was also present suggests that FtsA FtsZ interactions have coevolved and that the residues which differ between the E. coli proteins and those of the two other species may be important for specific interactions. PMID- 9352932 TI - Origin and evolution of group I introns in cyanobacterial tRNA genes. AB - Many tRNA(Leu)UAA genes from plastids contain a group I intron. An intron is also inserted in the same gene at the same position in cyanobacteria, the bacterial progenitors of plastids, suggesting an ancient bacterial origin for this intron. A group I intron has also been found in the tRNA(fMet) gene of some cyanobacteria but not in plastids, suggesting a more recent origin for this intron. In this study, we investigate the phylogenetic distributions of the two introns among cyanobacteria, from the earliest branching to the more derived species. The phylogenetic distribution of the tRNA(Leu)UAA intron follows the clustering of rRNA sequences, being either absent or present in clades of closely related species, with only one exception in the Pseudanabaena group. Our data support the notion that the tRNA(Leu)UAA intron was inherited by cyanobacteria and plastids through a common ancestor. Conversely, the tRNA(fMet) intron has a sporadic distribution, implying that many gains and losses occurred during cyanobacterial evolution. Interestingly, a phylogenetic tree inferred from intronic sequences clearly separates the different tRNA introns, suggesting that each family has its own evolutionary history. PMID- 9352933 TI - The Helicobacter pylori genome is modified at CATG by the product of hpyIM. AB - To understand mechanisms of DNA methylation in Helicobacter pylori, a human pathogen associated with peptic ulcer disease and gastric adenocarcinoma, we cloned a putative DNA methyltransferase gene, hpyIM. This gene contains a 990-bp open reading frame encoding a 329-amino-acid protein, M.HpyI. Sequence analysis revealed that M.HpyI was closely related to CATG-recognizing adenine DNA methyltransferases, including M.NlaIII in N. lactamica. hpyIM was present in all H. pylori strains tested. DNA from wild-type H. pylori strains was resistant to digestion by SphI and NlaIII, which recognize DNA at sites containing CATG, whereas their isogenic hpyIM mutants were susceptible, indicating lack of modification. Overexpression of hpyIM in Escherichia coli rendered DNA from these cells resistant to NlaIII digestion, confirming the role of hpyIM in modifying CATG sites. We conclude that hpyIM encodes a DNA methyltransferase, M.HpyI, that is well conserved among diverse H. pylori strains and that modifies H. pylori genomes at CATG sites. PMID- 9352934 TI - The tigA gene is a transcriptional fusion of glycolytic genes encoding triose phosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase in oomycota. AB - Genes encoding triose-phosphate isomerase (TPI) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are fused and form a single transcriptional unit (tigA) in Phytophthora species, members of the order Pythiales in the phylum Oomycota. This is the first demonstration of glycolytic gene fusion in eukaryotes and the first case of a TPI-GAPDH fusion in any organism. The tigA gene from Phytophthora infestans has a typical Oomycota transcriptional start point consensus sequence and, in common with most Phytophthora genes, has no introns. Furthermore, Southern and PCR analyses suggest that the same organization exists in other closely related genera, such as Pythium, from the same order (Oomycota), as well as more distantly related genera, Saprolegnia and Achlya, in the order Saprolegniales. Evidence is provided that in P. infestans, there is at least one other discrete copy of a GAPDH-encoding gene but not of a TPI-encoding gene. Finally, a phylogenetic analysis of TPI does not place Phytophthora within the assemblage of crown eukaryotes and suggests TPI may not be particularly useful for resolving relationships among major eukaryotic groups. PMID- 9352935 TI - Expression of a streptomycete leaderless mRNA encoding chloramphenicol acetyltransferase in Escherichia coli. AB - The chloramphenicol acetyltransferase (cat) gene from Streptomyces acrimycini encodes a leaderless mRNA. Expression of the cat coding sequence as a leaderless mRNA from a modified lac promoter resulted in chloramphenicol resistance in Escherichia coli. Transcript mapping with nuclease S1 confirmed that the 5' end of the cat message initiated at the A of the AUG translational start codon. Site directed mutagenesis of the lac promoter or the cat start codon abolished chloramphenicol resistance, indicating that E. coli initiated translation at the 5' terminal AUG of the cat leaderless mRNA. Addition of 5'-AUGC-3' to the 5' end of the cat mRNA resulted in translation occurring also from the reading frame defined by the added AUG triplet, suggesting that a 5'-terminal start codon is an important recognition feature for initiation and establishing reading frame during translation of leaderless mRNA. Addition of an untranslated leader and Shine-Dalgarno sequence to the cat coding sequence increased cat expression in a cat:lacZ fusion; however, the level of expression was significantly lower than when a fragment of the bacteriophage lambda cI gene, also encoding a leaderless mRNA, was fused to lacZ. These results indicate that in the absence of an untranslated leader and Shine-Dalgarno sequence, the streptomycete cat mRNA is translated by E. coli; however, the cat translation signals, or other features of the cat mRNA, provide for only a low level of expression in E. coli. PMID- 9352936 TI - Catalase-peroxidase of Caulobacter crescentus: function and role in stationary phase survival. AB - Caulobacter crescentus is an obligate aerobe which is exposed to high concentrations of photosynthetic oxygen and low levels of nutrients in its aquatic environment. Physiological studies of oxidative and starvation stresses in C. crescentus were undertaken through a study of lacZ fusion and null mutant strains constructed from the cloned 5' end of katG, encoding a catalase peroxidase. The katG gene was shown to be solely responsible for catalase and peroxidase activity in C. crescentus. Like the katG of Escherichia coli, C. crescentus katG is induced by hydrogen peroxide and is important in sustaining the exponential growth rate. However, dramatic differences are seen in growth stage induction. E. coli KatE catalase and KatG catalase-peroxidase activities are induced 15- to 20-fold during exponential growth and then approximately halved in the stationary phase. In contrast, C. crescentus KatG activity is constant throughout exponential growth and is induced 50-fold in the stationary phase. Moreover, the survival of a C. crescentus katG null mutant is reduced by more than 3 orders of magnitude after 24 h in stationary phase and more than 6 orders of magnitude after 48 h, a phenotype not seen for E. coli katE and katG null mutants. These results indicate a major role for C. crescentus catalase peroxidase in stationary-phase survival and raise questions about whether the peroxidatic activity as well as the protective catalatic activity of the dual function enzyme is important in the response to starvation stress. PMID- 9352937 TI - Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene. AB - A transient chloramphenicol acetyltransferase (CAT) expression system was developed for Borrelia burgdorferi. An Escherichia coli vector containing a promoterless Streptococcus agalactiae cat gene was constructed. Promoters for ospA, ospC, and flaB were placed upstream of this cat gene, and CAT assays were performed in E. coli from these stably maintained plasmids. The plasmids with putative promoters ospA and flaB were found to be approximately 20-fold more active than were the plasmids with ospC or no promoter. The level of activity correlated well with the resistance to chloramphenicol that each plasmid provided. Next, the nonreplicative plasmid constructs were transformed by electroporation into B. burgdorferi. CAT assays were performed by both thin-layer chromatography and the fluor diffusion method. Measurement of CAT activity demonstrated that the ospA promoter was again about 20-fold more active than the promoterless cat gene. The flaB and ospC promoters increased the activity seven- and threefold, respectively, over that with the promoterless construct. This simple transient-expression assay was shown to be an effective method to study promoter function in B. burgdorferi in the absence of a well-developed genetic system. PMID- 9352938 TI - The tyrocidine biosynthesis operon of Bacillus brevis: complete nucleotide sequence and biochemical characterization of functional internal adenylation domains. AB - The cyclic decapeptide antibiotic tyrocidine is produced by Bacillus brevis ATCC 8185 on an enzyme complex comprising three peptide synthetases, TycA, TycB, and TycC (tyrocidine synthetases 1, 2, and 3), via the nonribosomal pathway. However, previous molecular characterization of the tyrocidine synthetase-encoding operon was restricted to tycA, the gene that encodes the first one-module-bearing peptide synthetase. Here, we report the cloning and sequencing of the entire tyrocidine biosynthesis operon (39.5 kb) containing the tycA, tycB, and tycC genes. As deduced from the sequence data, TycB (404,562 Da) consists of three modules, including an epimerization domain, whereas TycC (723,577 Da) is composed of six modules and harbors a putative thioesterase domain at its C-terminal end. Each module incorporates one amino acid into the peptide product and can be further subdivided into domains responsible for substrate adenylation, thiolation, condensation, and epimerization (optional). We defined, cloned, and expressed in Escherichia coli five internal adenylation domains of TycB and TycC. Soluble His6-tagged proteins, ranging from 536 to 559 amino acids, were affinity purified and found to be active by amino acid-dependent ATP-PPi exchange assay. The detected amino acid specificities of the investigated domains manifested the colinear arrangement of the peptide product with the respective module in the corresponding peptide synthetases and explain the production of the four known naturally occurring tyrocidine variants. The Km values of the investigated adenylation domains for their amino acid substrates were found to be comparable to those published for undissected wild-type enzymes. These findings strongly support the functional integrities of single domains within multifunctional peptide synthetases. Directly downstream of the 3' end of the tycC gene, and probably transcribed in the tyrocidine operon, two tandem ABC transporters, which may be involved in conferring resistance against tyrocidine, and a putative thioesterase were found. PMID- 9352939 TI - Cloning of a Vibrio alginolyticus rpoN gene that is required for polar flagellar formation. AB - A fragment of DNA was cloned which complemented a polar flagellum-defective (pof) mutation of Vibrio alginolyticus. The fragment contained two complete and two partial open reading frames (ORFs) (ORF2 and -3 and ORF1 and -4, respectively). The presumed product of ORF2 has an amino acid sequence with a high degree of similarity to that of RpoN, which is an alternative sigma factor (sigma54) for other microorganisms. The other ORFs are also homologous to the genes adjacent to other rpoN genes. Deletion analysis suggests that ORF2 complements the pof mutation. These results demonstrate that RpoN is involved in the expression of polar flagellar genes. PMID- 9352940 TI - The acrAB homolog of Haemophilus influenzae codes for a functional multidrug efflux pump. AB - Disruption of gene HI0894 or HI0895 in Haemophilus influenzae Rd, homologs of Escherichia coli acrAB multidrug efflux genes, caused hypersusceptibility to erythromycin, rifampin, novobiocin, and dyes such as ethidium bromide and crystal violet and increased accumulation of radioactive erythromycin, showing that these genes are expressed and contribute to the baseline level resistance of this organism through active drug efflux. The gene disruption did not produce detectable changes in susceptibility to several other antibiotics, possibly because rapid influx of small antibiotic molecules through the large H. influenzae porin channels counterbalances their efflux. PMID- 9352941 TI - Inactivation of mdoH leads to increased expression of colanic acid capsular polysaccharide in Escherichia coli. AB - Capsule gene (cps) expression, which normally occurs at low levels in Escherichia coli lon+ cells, increased 38-fold in lon+ cells carrying a Tn10::delta kan insertion mapping to 24 min on the E. coli chromosome. Null mutations in rcsA, rcsB, or rcsC abolished the effect of the Tn10::delta kan insertion. Sequencing of both sides of the Tn10::delta kan insertion localized the insertion to the previously reported mdoH gene, which encodes a protein involved in biosynthesis of membrane-derived oligosaccharides (MDOs). A model suggesting that the periplasmic levels of MDOs act to signal RcsC to activate cps expression is proposed. PMID- 9352942 TI - SigmaE is an essential sigma factor in Escherichia coli. AB - SigmaE is an alternative sigma factor that controls the extracytoplasmic stress response in Escherichia coli. SigmaE is essential at high temperatures but was previously thought to be nonessential at temperatures below 37 degrees C. We present evidence that sigmaE is an essential sigma factor at all temperatures. Cells lacking sigmaE are able to grow at low temperatures because of the presence of a frequently arising, unlinked suppressor mutation. PMID- 9352943 TI - cis-Acting sequences required for light-responsive expression of the psbDII gene in Synechococcus sp. strain PCC 7942. AB - We analyzed the sequences required for promoter activity and high-light responsiveness of the psbDII gene in the cyanobacterium Synechococcus sp. strain PCC 7942 by using transcriptional fusions to a lacZ reporter gene. The basal promoter drives high constitutive expression, although no canonical -35 element is evident. The smallest fragment that showed clear light-responsive expression extends from -38 to +160, which includes 52 bp of the psbDII open reading frame. Sequences downstream from the promoter, within the untranslated leader region from +11 to +24, were required for high-light induction. PMID- 9352945 TI - Minimally invasive coronary artery surgery: the last operation. AB - Left anterior descending grafting with a left internal thoracic artery on a beating heart via a small left anterior thoracotomy is a procedure that is becoming popular, even if not yet standardized. From November 21, 1994 through February 20, 1997, 411 patients underwent a small left anterior thoracotomy; 206 had single-vessel disease, 205 had multiple-vessel disease. The early mortality rate was 1.0% (4 patients); causes of death were cardiac, not operation-related in 3, and non-cardiac in 1. The late mortality rate was 1.4% (6 patients); causes of death were cardiac operation-related in 1, non-cardiac in 3. All patients had a postoperative Doppler-flow velocity assessment; 231 (56.2%) underwent an angiographic control during the first postoperative year. Some patients were selected, as every patient with conduit or anastomotic malfunction underwent angiography. The patency rate was 92.4% (214/231); perfect distal anastomoses were obtained in 87.0% (201/231). With increasing experience and new instruments for left internal thoracic artery harvesting and left anterior descending artery stabilization, from April 21, 1996, patency rate increased to 98.2% (107/109) and perfect patency rate to 95.4% (104/109); results are therefore improving with time. The left anterior small thoracotomy procedure gives acceptable midterm results and is a reasonable alternative to the median sternotomy when the left anterior descending artery needs to be grafted with the left internal thoracic artery. PMID- 9352946 TI - Port-Access coronary artery bypass grafting. AB - New techniques for minimally invasive cardiac surgery have recently emerged. This report describes the Port-Access technique for coronary artery bypass grafting, which involves a small left anterior thoracotomy, femoral cannulation for endovascular cardiopulmonary bypass, and cardioplegic arrest using an endoaortic occlusion catheter and cardioplegia delivery system. This technique allows for minimally invasive single or multivessel revascularization in an arrested, protected heart, while maintaining a high level of anastomotic precision. The Port-Access surgical techniques are described, along with the indications and contraindications for this procedure. The initial New York University clinical results with Port-Access coronary bypass grafting are presented. PMID- 9352947 TI - Minimally invasive mitral valve surgery. AB - Because of advances in video-assisted general and thoracic surgery, minimally invasive cardiac surgery has been successfully performed experimentally and clinically. Recently described techniques of less invasive mitral valve surgery include limited right thoracotomy, parasternal incision, and partial sternotomy. These methods have been coupled to video-assisted thoracoscopy to further decrease the incision size. Cardiopulmonary bypass (central or peripheral) and either hypothermic fibrillatory arrest or cardioplegic arrest are used. The Port Access approach is a catheter-based system that provides effective cardiopulmonary bypass, cardioplegic arrest, and ventricular decompression. At Stanford University, 10 Port-Access mitral valve procedures were performed between May 1996 and January 1997. The mean age of the patients (eight men and two women) was 54 +/- 7 (SD) years. Nine patients had severe mitral regurgitation from myxomatous degeneration, and one suffered from severe mitral regurgitation and moderate mitral stenosis from a rheumatic etiology. Five patients underwent mitral valve replacement, and five underwent mitral valve repair. There was no operative mortality. The mean incision length was 8.1 +/- 2.5 cm. The aortic "cross-clamp" time was 99 +/- 22 minutes, and the cardiopulmonary bypass time was 151 +/- 52 minutes. The total hospitalization averaged 4.3 +/- 1.4 days. One patient developed third-degree atrioventricular block, requiring a prolonged stay in the intensive care unit and pacemaker placement; the same patient was found to have a perivalvular leak on follow-up, requiring reoperation at 3 months. Port Access mitral valve procedures can be performed safely with satisfactory outcome. Greater clinical experience and long-term follow-up are necessary to fully assess these less invasive techniques of mitral valve surgery. PMID- 9352948 TI - Minimally invasive aortic valve replacement. AB - Aortic valve replacement has proven reliable, relieves life-threatening symptoms, and improves long-term survival of patients with aortic stenosis and aortic regurgitation. Minimally invasive aortic valve replacement uses small incisions; reduces exposure of the patient to surgical trauma, blood utilization, and operative dissection; although still using cardiopulmonary bypass and achieving the same general quality as with the open operation. Early and medium term results for minimally invasive aortic valve replacement approaches show a reduction in pain, improved patient satisfaction, and improved mobility and return to full-time activity. Concomitantly, there should be decreased cost and a decreased reliance on post-hospital rehabilitation. PMID- 9352949 TI - Minimally invasive techniques for congenital heart surgery. AB - Minimally invasive techniques in congenital heart surgery have evolved steadily over the past 5 years. Initially, instrumentation and techniques were adopted from other subspecialties, and efforts were directed at simple extracardiac repairs. These efforts established the safety and efficacy of video-assisted endoscopic techniques for pediatric cardiac surgery. As instruments and techniques evolved, intraoperative cardioscopy became feasible and showed the utility of these new imaging techniques in facilitating open cardiac repairs by exposing remote areas within the heart. This experience has laid a foundation for the next phase of minimally invasive pediatric cardiac surgery: the repair of complex congenital defects. PMID- 9352950 TI - Diffuse malignant pleural mesothelioma: introduction. PMID- 9352951 TI - Pathology of diffuse malignant pleural mesothelioma. AB - An accurate diagnosis is essential to a rational approach to the treatment of diffuse malignant pleural mesothelioma and generally requires pathological examination with the application of special techniques. In recent years, immunohistochemistry has greatly abetted the distinction of mesothelioma from its many morphological mimics, yet diagnostic difficulties still remain because reactive hyperplasias and diverse tumors closely mimic mesothelioma. Mesotheliomas are classified into epithelial, mixed, sarcomatoid and undifferentiated types, based on conventional histological examination. The classification provides important prognostic information. Furthermore, differential diagnosis is directly related to histological type. Although such special techniques as histochemistry and electron microscopy continue to play an important role in some cases, immunohistochemistry often has replaced these in distinguishing epithelial-type mesothelioma from metastatic adenocarcinoma. It is also helpful in distinguishing sarcomatoid mesothelioma from it numerous morphological mimics. The distinction of mesothelioma from reactive mesothelial proliferations is still based on morphological examination and may be quite problematic. Recent cytogenetic studies, which have identified characteristic clonal deletions in mesotheliomas, give promise of providing valuable assistance in this distinction in the future. PMID- 9352952 TI - Surgical staging and work-up of patients with diffuse malignant pleural mesothelioma. AB - The current modalities used to treat diffuse malignant pleural mesothelioma (DMPM) have not been evaluated in the setting of prospective, multi-institutional randomized trials for two reasons: DMPM is a rare disease, and there is a lack of a widely accepted and standarized staging system. Several staging systems have been proposed in an effort to categorize patients with DMPM into homogeneous groups. These efforts have been hampered by a lack of correlation between staging and survival. In our institutional experience, the Brigham staging system has been able to stratify patients with similar survival. This is an institutional experience that needs validation in a multi-institutional setting and, furthermore, in a trial based on stage-specific adjuvant therapies. PMID- 9352953 TI - Chemotherapy for malignant mesothelioma. AB - The treatment of malignant mesothelioma (MM) has been challenging. Many series from larger single institutions comprise small numbers of selected patients. Positive studies tend to be published, whereas publication of negative studies is delayed, appears in obscure journals, or does not occur at all. Because of the pleural distribution of the tumor, reliable determination of response is problematic. Finally, the natural history of MM is generally short, but can be quite variable. Nevertheless, doxorubicin, cisplatin, and ifosfamide and perhaps other agents as well have modest activity. Larger phase II studies are now being done by cooperative groups accruing patients from community hospitals as well as from tertiary care centers. In the Cancer and Leukemia Group B study, the response rate in patients with measurable disease was 24% for cisplatin and either mitomycin C and doxorubicin, the highest response rates reported for a cooperative group study. Survival was slightly better for the doxorubicin combination. Studies of new drugs and biologics as well as of novel methods of drug delivery are underway. PMID- 9352954 TI - Pleurectomy/decortication in the setting of multimodality treatment for diffuse malignant pleural mesothelioma. AB - Pleurectomy/decortication is a frequently performed operation for patients with diffuse malignant pleural mesothelioma (DMPM). It has a low surgical mortality rate (less than 5%), but is associated with a significant risk of local recurrence. To date, intensive adjuvant radiation or chemotherapy has not diminished that risk. Despite these disappointing results, pleurectomy/decortication may still be the best treatment option for some patients, particularly those with early stage disease whose medical condition precludes pneumonectomy. The role of pleurectomy/decortication in conjunction with newer treatment strategies such as neoadjuvant therapy or gene therapy warrants investigation. PMID- 9352955 TI - Extrapleural pneumonectomy in the setting of multimodality therapy for diffuse malignant pleural mesothelioma. AB - Diffuse malignant pleural mesothelioma, a rare disease, is characterized by an aggressive local behavior and scant response to therapy. The first series using single modality therapy showed failure in terms of survival and local control. More recently, multimodality therapy has been used against this disease with better results, but still with more room for substantial improvement. The current multimodality series reported are isolated, single-institutional experiences with different treatment schemes, using different staging systems, most of which have not been validated. There is an enormous need for multiinstitutional prospective trials to evaluate the current treatment schemes in light of the steady increase in the incidence of this lethal tumor. The trimodality therapy used at the Brigham and Women's Hospital for selected patients is described. PMID- 9352956 TI - New therapies in the treatment of malignant pleural mesothelioma. AB - It can be safely stated that currently there exists no standard therapy for malignant mesothelioma. The "standard" methods of chemotherapy, radiation therapy, and surgery have all been used with little improvement in overall survival. Trimodality therapy that involves a combination of all three standard treatment modalities has been shown to improve survival in selected patients. New and innovative treatment strategies clearly are needed for a disease which, because of the disappointment with standard therapy, is most commonly approached with only palliative intent. The location of this malignancy and its tendency to remain localized make it an ideal target for intracavitary approaches using photodynamic therapy, targeted cytokines, and gene therapy. Strategies using modulation of the immune system in an attempt to elicit a specific response to the tumor have been combined with chemotherapy to optimize response. Lessons learned from treating this localized malignancy with novel therapies may have much broader implications for other tumors in which systemic disease predominates. PMID- 9352957 TI - Monitoring the safety of herbal medicines. AB - Extremely limited knowledge about the ingredients in herbal medicines and their effects in humans, the lack of stringent quality control and the heterogenous nature of herbal medicines all necessitate the continuous monitoring of the safety of these products. In Hong Kong, safety information on herbal medicines has come from the enquiries and reports received by our Drug and Poisons Information Service, on-going surveillance of patients treated in a large general teaching hospital and review of reports from the medical literature. Circumstances under which poisonings have occurred are also analysed in order to devise preventive measures. Once collected, this information is then distributed to health professionals in Hong Kong and abroad. WHO projects and pilot studies in Europe are also under way to promote and facilitate reporting of adverse reactions to herbal medicines. PMID- 9352958 TI - The challenge of effectively communicating risk-benefit information. AB - Although the techniques involved in drug safety monitoring (pharmacovigilance) have dramatically improved in recent years, communication of these issues to health professionals and the public lags far behind. Several measures need to be taken in order to address this discrepancy. A climate of greater openness concerning the basis of merit assessments must be created. We need to develop merit-assessment formulations that are more accurate and helpful when treating individual patients in clinical situations. All of the involved groups must be educated about the nature of drugs and drug therapy, and the possibilities and limitations of such therapy. More effective techniques and systems have to be developed in order to stimulate higher rates of high quality spontaneous reporting of adverse effects. More conscientious and purposeful attention to the theory and practice of communications, in order to ensure the effective delivery of optimal benefits to patients, clinicians and society at large, would also be advantageous. We must ensure that where issues of public health and confidence in the medical profession are at stake, we employ the very best communications practices. PMID- 9352959 TI - Newer anticonvulsant drugs: role of pharmacology, drug interactions and adverse reactions in drug choice. AB - In the last few years a number of new anticonvulsants have been introduced into clinical practice mainly as add-on therapy in patients who do not become seizure free while receiving established anticonvulsants. Up to now, no single drug has been shown to be more effective at controlling seizures of a particular type than another, so other factors such as mechanism of action, pharmacokinetics, dosage regimens or the spectrum of adverse drug reactions and interactions are used when making a choice between one agent and another. The mechanism of action of tiagabine and vigabatrin is very specific; both agents increase gamma aminobutyric acid (GABA) levels through inhibition of reuptake and catabolism respectively. However, the mechanism of action of gabapentin is unknown and those of felbamate, lamotrigine and topiramate are not sufficiently clarified as yet, and may be multiple. Great advances have been made in improving the pharmacokinetic characteristics of these newer anticonvulsants. Gabapentin and vigabatrin exhibit relatively ideal pharmacokinetic properties as they are not bound to proteins, are excreted mostly unchanged in the urine and show linear pharmacokinetics. Lamotrigine possesses a highly variable elimination half-life depending on the co-medication. Tiagabine is highly protein bound and zonisamide shows nonlinear pharmacokinetics; both these drugs are extensively metabolised. Problematic drug interactions between newer anticonvulsants and other drugs in general occur rarely when these agents are given concomitantly. However, in common with most new drugs, there are very few data on the use of the newer anticonvulsants in women of childbearing age. Studies done so far on interactions with oral contraceptives used low anticonvulsant dosages for a very short time. The newer anticonvulsants elicit adverse reactions that, while not being unique, are particularly associated with that drug. For example, felbamate may cause aplastic anaemia and fulminant liver failure, lamotrigine is prone to cause skin rash, and oxcarbazepine may cause symptomatic hyponatraemia. Topiramate and zonisamide cause kidney stones, and vigabatrin may induce psychiatric syndromes. Although highly diverse in structure and activity, these newer drugs offer new possibilities for treating refractory epilepsy. However, since no single factor can dictate the choice of drug nor predict the success of treatment, prescribing of these rather expensive drugs has to depend upon careful consideration of the aims of treatment, the characteristics of the drug and the needs of the individual patient. PMID- 9352961 TI - A risk-benefit assessment of mirtazapine in the treatment of depression. AB - Mirtazapine is the first of a new class of antidepressants, the noradrenergic and specific serotonergic antidepressants (NaSSA). Its antidepressant effect appears to be related to its dual enhancement of central noradrenergic and serotonin 5 HT1 receptor-mediated serotonergic neurotransmission. Mirtazapine possesses a number of useful pharmacokinetic characteristics such as good absorption, linear pharmacokinetics over the recommended dosage range (15 to 80 mg/day), and an elimination half-life of 20 to 40 hours, thereby allowing once-daily administration. However, since the drug is extensively metabolised by the hepatic cytochrome P450 (CYP) system and is excreted mainly in the urine, its clearance may be reduced by hepatic or renal impairment. In vitro data suggest that from a clinical point of view it is unlikely that mirtazapine would inhibit the metabolism of coadministered drugs metabolised by CYP1A2, CYP2D6 or CYP3A4. In vivo data from a study in extensive and poor metabolisers of debrisoquine indicate that strong inhibitors of CYP2D6 would have no effect on the concentration of racemic mirtazapine. In some placebo-controlled studies mirtazapine showed an early onset of antidepressant action, with significant reductions in total Hamilton Depression Rating Scale and Montgomery-Asberg Depression Rating Scale scores (relative to placebo) noted as early as 1 week after starting treatment. This therapeutic advantage was subsequently maintained during treatment, with mirtazapine proving significantly superior to placebo at treatment end-point in the majority of studies. In comparative trials, the antidepressant efficacy of mirtazapine was comparable with that of tricyclic antidepressants such as amitriptyline, clomipramine and doxepin, and in 2 studies superior to that of trazodone and fluoxetine. Mirtazapine appears to have a broad spectrum of activity, reflected in its efficacy in a variety of clinical settings. Its additional beneficial effects on the symptoms of anxiety and sleep disturbance associated with depression may reduce the need for concomitant anxiolytic and hypnotic medication seen with some antidepressants. Mirtazapine has demonstrated superior tolerability to the tricyclic antidepressants and trazodone, primarily on account of its relative absence of anticholinergic, adrenergic and serotonin-related adverse effects, in particular gastrointestinal adverse effects and sexual dysfunction. It appears that increased sedation associated with the drug is related to subtherapeutic dosages, and that it is reported in substantially fewer patients when the drug is used in appropriate dosages (> or = 15 mg as a single evening dose) from the beginning of treatment. Although 2 cases of reversible severe symptomatic neutropenia have been reported in clinical trials, there have been no additional reports of symptomatic neutropenia since the introduction of this drug to various countries in September 1994. Currently available data and initial clinical experience suggest that with its combination of dual action, simple pharmacokinetics, and clinical efficacy and tolerability, mirtazapine appears to be an important advance in the pharmacotherapy of depression. PMID- 9352960 TI - Safety aspects of parenteral iron in patients with end-stage renal disease. AB - Absolute and functional iron deficiency is the most common cause of epoetin (recombinant human erythropoietin) hyporesponsiveness in renal failure patients. Diagnostic procedures for determining iron deficiency include measurement of serum iron levels, serum ferritin levels, saturation of transferrin and percentage of hypochromic red blood cells. Patients with iron deficiency should receive supplemental iron, either orally or intravenously. Adequate intravenous iron supplementation allows reduction of epoetin dosage by approximately 40%. Intravenous iron supplementation is recommended for all patients undergoing haemodialysis and for pre-dialysis and peritoneal dialysis patients with severe iron deficiency. During the maintenance phase (period of epoetin therapy after correction of iron deficiency), the use of low-dose intravenous iron supplementation (10 to 20 mg per haemodialysis treatment or 100 mg every second week) avoids iron overtreatment and minimises potential adverse effects. Depending on the degree of pre-existing iron deficiency, markedly higher iron doses are necessary during the correction phase (period of epoetin therapy after correction of iron deficiency) [e.g. intravenous iron 40 to 100 mg per haemodialysis session up to a total dose of 1000 mg]. The iron status should be monitored monthly during the correction phase and every 3 months during the maintenance phase to avoid overtreatment with intravenous iron. PMID- 9352963 TI - Documentation of decline in morbidity in women undergoing coronary angioplasty (a report from the 1993-94 NHLBI Percutaneous Transluminal Coronary Angioplasty Registry). National Heart, Lung, and Blood Institute. AB - To determine whether there has been an improvement in the relatively unfavorable outcome of percutaneous transluminal coronary angioplasty (PTCA) in women, the 1993 to 1994 National Heart, Lung, and Blood Institute Percutaneous Transluminal Coronary Angioplasty Registry collected data from 12 clinical centers that participated in the earlier registries. We compared 274 consecutive women in 1993 to 1994 with 545 consecutive women in 1985 to 1986 undergoing PTCA. Women in the 1993 to 1994 registry were older (64.3 vs 61.0 years, p <0.001) with more diabetes mellitus (34.3% vs 19.9%, p <0.001), congestive heart failure (13.7% vs 8.6%, p <0.05), and comorbid disease (19.5% vs 9.3%, p <0.001). Left ventricular function and multivessel coronary artery disease were similar between groups. Angiographic success (90.9% vs 85.1%, p <0.05) and clinical success (89.4% vs 79.4%, p <0.001) were higher in women undergoing PTCA in 1993 to 1994 than in 1985 to 1986. Whereas there was no difference in in-hospital mortality (1.5% vs 2.6%), the incidence of nonfatal myocardial infarction (1.8% vs 4.6%, p <0.05), emergency coronary artery bypass graft surgery (1.8% vs 4.6%, p <0.05), and the combined end points of death, myocardial infarction, and emergency coronary artery bypass grafting (4.4% vs 9.7%, p <0.01) were lower in women in 1993 to 1994 than in women in 1985 to 1986, respectively. Multivariate analysis revealed an odds ratio of 0.36 (95% confidence interval 0.18 to 0.72) for major complications and of 2.34 (95% confidence interval, 1.49 to 3.69) for clinical success in the 1993 to 1994 versus 1985 to 1986 registry. Therefore, despite a higher risk profile, women undergoing PTCA in 1993 to 1994 have a higher clinical success and lower major complication rate than women treated with PTCA in 1985 to 1986. PMID- 9352964 TI - Effect of platelet glycoprotein IIb/IIIa receptor inhibition on distal embolization during percutaneous revascularization of aortocoronary saphenous vein grafts. EPIC Investigators. Evaluation of IIb/IIIa platelet receptor antagonist 7E3 in Preventing Ischemic Complications. AB - Percutaneous treatment of narrowed aortocoronary saphenous vein graft disease represents a viable option for patients with recurrent angina following coronary artery bypass grafting. Present strategies are limited by high rates of distal embolization, non-Q-wave acute myocardial infarction (AMI), and restenosis. Because these complications may be mediated by platelets, inhibition of platelet glycoprotein IIb/IIIa receptor, the final common pathway for aggregation, may improve clinical outcomes. In the Evaluation of IIb/IIIa platelet receptor antagonist 7E3 in Preventing Ischemic Complications (EPIC) trial, 2,099 patients undergoing high-risk percutaneous coronary revascularization were randomized to receive abciximab bolus and infusion, abciximab bolus followed by placebo infusion or placebo. A total of 101 patients were treated for narrowing of saphenous vein grafts, 38 in the bolus and infusion group, 34 in the bolus group and 29 in the placebo group. Clinical end points included all-cause mortality, nonfatal AMI and need for repeat revascularization at 30 days. Compared with placebo, bolus and infusion therapy resulted in a significant reduction in distal embolization (2% vs 18%, p = 0.017) and a trend towards reduction in early large non-Q-wave AMI (2% vs 12%, p = 0.165). The occurrence of a 30-day composite end point was similar among the 3 treatment groups. At 6 months, there was also no difference in the composite end point. These results suggest that adjunctive therapy with abciximab during percutaneous treatment of narrowed saphenous vein grafts reduces the occurrence of distal embolization, and possibly non-Q-wave AMI. PMID- 9352962 TI - Drug-induced cardiac arrhythmias: incidence, prevention and management. AB - Drugs can cause cardiac arrhythmias in a number of clinical situations, and many of the implicated agents are used to treat non-cardiac conditions. These adverse effects are frequently idiosyncratic, but are often mediated via triggered activity causing torsade de pointes. Drugs being used for treatment of cardiac conditions may promote arrhythmias by re-entrant mechanisms or via triggered activity. Many drugs may cause cardiac arrhythmic complications when taken in excessive amounts. Keys to reducing the incidence of drug-induced cardiac arrhythmias include increased awareness among the medical, pharmaceutical and nursing professions of the potential problems in using certain agents, especially in specific situations. Appropriate monitoring when such treatment is essential and, after diagnosis, prompt withdrawal of the offending agent and treatment for the arrhythmia should be initiated. PMID- 9352965 TI - Usefulness of respiratory gated magnetic resonance coronary angiography in assessing narrowings > or = 50% in diameter in native coronary arteries and in aortocoronary bypass conduits. AB - Magnetic resonance coronary angiography (MRCA) is a promising method for the assessment of proximal coronary artery stenosis. Conventional 2-dimensional techniques require repetitive breath holds to image multiple sections. This may lead to misregistrations if the respiratory level is not exactly reproduced. In the present study, MRCA was performed using a 3-dimensional approach with navigator echo-based respiratory gating. In 73 patients (55 men and 18 women) who were referred for cardiac catheterization, the assessment of significant stenoses (> or = 50%) was performed in the proximal and midsegments of the coronary arteries after multiplanar reconstruction of the visualized coronary arteries. In addition, in 8 patients with coronary artery bypass grafts the patency of the transplants was evaluated. After withdrawing 8 patients from analysis because of poor image quality, stenosis evaluation was possible in 236 of 455 reviewed coronary segments (52%). In the other 219 cases, either the visualization of the vessel segment was indistinct (30%) or the segment was located outside the imaging volume (18%). In total, 28 of 43 significant coronary stenoses could be correctly identified (65%). Evaluation of bypass graft patency was possible in 7 patients. All 4 occluded and 13 of 15 patent grafts were correctly classified. Thus, respiratory gated MRCA is a feasable method for the assessment of hemodynamically significant coronary stenoses and bypass graft patency. However, technical improvements are mandatory to improve accuracy of the method. PMID- 9352966 TI - Incidence and angiographic predictors of side branch occlusion following high pressure intracoronary stenting. AB - We evaluated the incidence, angiographic predictors, and clinical outcome of side branch occlusion (SBO) following high-pressure intracoronary stenting in 175 patients. All stent implants during a 7-month period were reviewed for the incidence of major (>1 mm) SBO. Side branches were further characterized based on side branch and index lesion morphology. Clinical events (death, myocardial infarction, and target vessel revascularization rates) were determined at 9 months. A total of 175 patients (182 lesions) had 224 major side branches covered by intracoronary stents. Of these, 43 (19%) occluded. Most SBOs (29 of 43 [67%]) occurred after poststent dilation using high-pressure inflations (15.3 +/- 3.3 atmospheres). No clinical characteristics correlated with SBO. By multivariate analysis, those side branches with >50% ostial narrowing that arose from within or just beyond the diseased portion of the parent vessel (threatened side branch morphologies) were a powerful angiographic predictor of SBO (odds ratio 40, 95% confidence interval, 14 to 130, p <0.0001). At 9-month follow-up there was no difference in combined clinical events between those patients with and without SBO. These data demonstrate that side branches with ostial stenoses in continuity with diseased parent lesions were at risk of occlusion following stenting. SBO, however, was not associated with adverse clinical outcome. These findings lend support to plaque shift ("snow plow effect") as the mechanism behind SBO following stent placement. PMID- 9352967 TI - Comparison of six-month outcome of coronary artery stenting in patients <65, 65 75, and >75 years of age. AB - We studied 1,238 patients receiving 1,880 coronary stents. In-hospital outcomes were divided by age into <65 years (n = 747, group 1), 65 to 75 years (n = 326, group 2), and >75 years (n = 165, group 3). Procedural success was 97.2%, 95.1%, and 98.8% in groups 1, 2, and 3, respectively (p = NS). There was 1 death (group 1). Myocardial infarction occurred in 1.2%, 2.8%, and 1.8%, bypass surgery occurred in 0.9%, 1.8%, and 1.2%, and repeat balloon angioplasty in 0.3%, 0.6%, and 0% of patients in groups 1, 2, and 3, respectively (p = NS for all comparisons). Vascular complications occurred in 2.8%, 4.9%, and 6.1% in groups 1, 2, and 3, respectively (p <0.05). Six-month follow-up of patients was divided by age: <65 years (n = 564, group 1); 65 to 75 years (n = 221, group 2); and >75 years (n = 122, group 3). Event-free survival was 94.5%, 90.5%, and 89.3% for groups 1, 2, and 3, respectively (p = NS). Death occurred in 0.4%, 0.5%, and 1.6%; myocardial infarction occurred in 1.2%, 2.3%, and 1.6%, and target vessel revascularization in 4.3%, 8.6%, and 7.4% for groups 1, 2, and 3, respectively (p = NS for all comparisons). Thus, coronary stenting produced favorable in-hospital and 6-month outcomes in all 3 age groups. Age itself should not preclude patients from undergoing coronary stenting. PMID- 9352968 TI - C-reactive protein elevation and early outcome in patients with unstable angina pectoris. AB - C-reactive protein, a reactant of the acute phase of inflammation, has been shown to be increased in patients with unstable angina. Moreover, it has recently been found that increased C-reactive protein is associated with a poor outcome during hospitalization in selected patients with severe unstable angina. The aim of this study was to investigate the prognostic value of C-reactive protein elevation in a large population with unstable angina. We measured serum levels of this marker in 140 patients hospitalized with unstable angina (class IIIB of the Braunwald classification, mean time from last anginal episode 5 +/- 5 hours). Thirty-nine of them (28%) had increased serum levels on hospital admission and 33 (24%) experienced an adverse outcome (myocardial infarction or refractory angina) during hospitalization. Kaplan-Meier analysis showed that the probability of developing cardiac events during hospitalization was not different between patients with and without abnormal C-reactive protein levels. Furthermore, the incidence of ischemia at Holter monitoring during the first 72 hours after hospitalization was not different between patients with and without abnormal C reactive protein. In a representative population of patients with unstable angina, a sizable proportion had increased serum C-reactive protein levels; however, abnormal concentrations of C-reactive protein do not predict an adverse outcome in the early phase after the acute episode. PMID- 9352969 TI - Effect of left ventricular function on the assessment of myocardial viability by technectium-99m sestamibi and correlation with positron emission tomography in patients with healed myocardial infarcts or stable angina pectoris, or both. AB - The accuracy of technetium-99m (Tc-99m) sestamibi single-photon emission computed tomography (SPECT) for the assessment of myocardial viability in patients with coronary artery disease and left ventricular (LV) dysfunction is not defined completely. This study determines whether the performance of Tc-99m sestamibi SPECT for viability detection differs between patients with mild-to-moderate coronary artery disease. Patients with regional and/or global LV dysfunction were separated into 2 groups on the basis of LV ejection fraction (EF) at rest: group 1 (LVEF > 25%, mean 36 +/- 6%, n = 9), and group 2 (LVEF < or = 25%, mean 17 +/- 5%, n = 11). All patients underwent semiquantitative Tc-99m sestamibi SPECT and positron emission tomography (PET) at rest with N-13 ammonia and F-18 fluorodeoxyglucose. The overall regional concordance of SPECT and PET for viability detection was 89% in group 1 and 78% in group 2 (p = 0.002). Discordance in group 2 was almost exclusively due to PET viable and/or SPECT nonviable regions. In regions with hypoperfusion at rest by PET, concordance was 78% in group 1 and only 64% in group 2 (p = 0.0015). In regions with reduced perfusion and relatively increased metabolic activity ("flow: metabolism mismatch"), Tc-99m sestamibi SPECT identified 88% of regions in group 1 as viable, but only 42% of regions in group 2 (p = 0.002). Thus, while Tc-99m sestamibi semiquantitative SPECT at rest shows a good concordance with PET for the detection of myocardial viability in patients with coronary artery disease with mild-to-moderate LV dysfunction, it may underestimate myocardial viability in patients with severe LV dysfunction, particularly in those patients with hypoperfusion at rest as assessed by PET. PMID- 9352970 TI - Safety and results of dobutamine stress echocardiography in women versus men and in patients older and younger than 75 years of age. AB - The purpose of this retrospective study was to examine 732 consecutive patients who underwent dobutamine stress echocardiography (DSE) in order to compare the safety and result profiles of this test between women versus men and in patients > or = 75 and < 75 years of age. Our study included 416 women (57%) and 316 men (43%; mean age 62 +/- 12 years [range 16 to 93]). Patients were divided into 3 age groups: (1) group I (n = 179): < 55 years (mean 47 +/- 6), (2) group II (n = 447): 55 to 74 years (mean 64 +/- 5), and (3) group III (n = 106): > or = 75 years (mean 80 +/- 4). DSE was more likely to have negative results in women than in men (prevalence of positivity = 20% vs 31%, p = 0.001), but DSE had a similar safety profile in both genders. Women required lower doses of dobutamine and atropine to reach an end point. There was a similar incidence of test positivity in older and younger patients (23% in group I, 24% in group II, and 30% in group III, p = NS). DSE was generally a safe test in patients > or = 75 years, but there was a different safety profile in the elderly group compared with younger patients--specifically, more frequent asymptomatic hypotension (7% in group I, 13% in group II, and 25% in group III, p = 0.0002) and ventricular arrhythmias (26% in group I, 30% in group II, and 41% in group III, p = 0.04), but less frequent chest pain (32% in group I, 23% in group II, and 17% in group III, p = 0.009). Multivariate analysis suggested that the baseline usage of beta blockers was also a major determinant of the safety and ischemia profile during DSE. In conclusion, there were significant gender- and/or age-specific differences in the safety and test result profile of DSE. These differences should be considered when performing or interpreting DSE, particularly in women and in patients aged > or = 75 years. PMID- 9352971 TI - Acute effects of conjugated estrogens on coronary blood flow response to acetylcholine in men. AB - Estrogen therapy is associated with a 50% reduction in the clinical manifestations of coronary artery disease in postmenopausal women. Attenuation of coronary vasomotor dysfunction may contribute to estrogen's cardioprotective effects. We hypothesized that conjugated estrogens, which contain several vasoactive estrogenic compounds, may favorably influence the vasomotor response to acetylcholine in men. Twenty men, 56 +/- 5 years of age, referred for clinically indicated coronary angiography, participated in this study. Acetylcholine-induced changes in coronary flow were measured by quantitative coronary angiography and intracoronary Doppler ultrasonography before and 15 minutes after intravenous administration of conjugated estrogens (0.625 mg) in 12 men and placebo in 8 men. Initial acetylcholine infusion resulted in no significant increase in coronary blood flow. However, 15 minutes after estrogen administration repeat acetylcholine infusion caused a mean 32% increase in coronary blood flow from 41 +/- 5 to 54 +/- 8 ml/min (p = 0.02). Acetylcholine induced change in flow after estrogen was significantly different from that before estrogen (p = 0.03). Placebo administration did not affect acetylcholine induced changes in coronary flow. Thus, intravenous conjugated estrogens favorably modulate acetylcholine-induced changes in coronary hemodynamics in men. This suggests that novel nonfeminizing estrogenic compounds may have anti ischemic effects in men. PMID- 9352972 TI - Skeletal muscle endurance training improves peripheral oxidative capacity, exercise tolerance, and health-related quality of life in women with chronic congestive heart failure secondary to either ischemic cardiomyopathy or idiopathic dilated cardiomyopathy. AB - Despite reported benefits of exercise training in men with chronic congestive heart failure (CHF) and in both men and women with coronary artery disease, the effects of training in women with CHF have not been throughly investigated. Therefore, 16 women (62 +/- 10 years [mean +/- SD]) with stable, moderate, chronic CHF (left ventricular ejection fraction 28 +/- 8%) were studied in a randomized crossover trial with 8 weeks of knee extensor endurance training and 8 weeks of nontraining. The effects of the exercise-based rehabilitation were assessed in skeletal muscle metabolic capacity, exercise tolerance, and quality of life. The compliance rate in training was 98% and no adverse events occurred during the study period. Training increased the activity of citrate synthase (44%, p <0.0001) and lactate dehydrogenase (23%, p <0.002) in the trained muscles, and an improved oxidative capacity in relation to the glycolytic capacity (23%, p <0.002) was found. Peak oxygen uptake (14%, p <0.0005) and peak work rate (43%, p <0.0001) during incremental exercise increased, and blood lactate concentration during standardized submaximal exercise and during the recovery phase decreased (17%, p <0.05). The distance ambulated during 6 minutes (p <0.03), and the overall (p <0.01), physical (p <0.05), and psychosocial (p <0.03) health-related quality of life improved. Because the skeletal muscle endurance training improved peripheral oxidative capacity, exercise tolerance, and the health-related quality of life without any adverse events, this mode of training can be recommended for women with chronic heart failure. PMID- 9352973 TI - Role of transthoracic and transesophageal echocardiography in predicting embolic events in patients with active infective endocarditis involving native cardiac valves. AB - Some studies describe an increased risk for emboli in infective endocarditis patients with large (>10 mm) and mobile vegetations. Other studies fail to demonstrate the above relation. Most studies have been performed using transthoracic echocardiography or with a monoplane transesophageal approach. The present study examines whether distinctive characteristics of vegetative lesions detected by transthoracic and multiplane transesophageal echocardiography are predictive of embolic risk. We reviewed both transthoracic and transesophageal echocardiograms of 57 patients with diagnosis of acute infective endocarditis and no documented or suspected previous embolic events. We evaluated site, length, width, mobility, and echodensity of vegetations. Twenty-five patients (44%) had embolic events. No statistical differences in age, sex distribution, location of endocarditis, or offending pathogens between embolic (n = 25) and nonembolic (n = 32) patients were found. There were no differences in any of the echo characteristics of vegetations detected by transthoracic and transesophageal approach in embolic and nonembolic groups. Thus, transthoracic and transesophageal characteristics of vegetations are not helpful in defining embolic risk in patients with infective endocarditis. PMID- 9352974 TI - Relation of systemic arterial pulse pressure to coronary atherosclerosis in patients with mitral stenosis. AB - The relation of a wide systemic arterial pulse pressure to coronary atherosclerosis has not been fully defined. One hundred fifty-nine patients > 40 years old with symptomatic mitral stenosis (MS) who received routine coronary angiography were classified into 2 groups according to the presence of > or = 50% diameter narrowing of > or = 1 coronary artery (n = 48) or no significant disease (n = 111). Pulse pressure was determined both by noninvasive sphygmomanometer and invasive catheterization methods. There were no significant differences in risk factors of coronary artery disease (CAD) or the severity of MS between the 2 groups. From multivariate logistic regression analysis, independent predictors of development of CAD in MS were age (standardized coefficient beta = 1.3437, p = 0.0025), gender (beta = 0.0107, p = 0.0105), mean blood pressure (beta = 1.1839, p = 0.0105), and pulse pressure (beta = 1.3157, p = 0.0008). A wide pulse pressure (> or = 60 mm Hg) correlated with the presence of angiographically significant CAD with a sensitivity and specificity of 88% and 77%. The negative predictive value was 93%. Pulse pressure assessed by sphygmomanometry provided important clinical information. A wide pulse pressure in patients with MS was associated with a high incidence of CAD. PMID- 9352975 TI - Identification of mitochondrial antigens recognized by antibodies in sera of patients with idiopathic dilated cardiomyopathy by two-dimensional gel electrophoresis and protein sequencing. AB - Antimitochondrial antibodies in sera of patients with idiopathic dilated cardiomyopathy (IDC) have been described previously, but the corresponding antigens have not been analyzed systematically. We therefore used both 1 dimensional and high-resolution 2-dimensional gel electrophoresis followed by immunoblotting and N-terminal amino acid sequencing to identify the relevant mitochondrial antigens, which are recognized by serum antibodies. Sera were obtained from patients with IDC (n = 75) and healthy controls (n = 182). For detection of antimitochondrial antibodies the mitochondrial antigen fraction, consisting of submitochondrial particles isolated from a bovine heart, was separated on SDS-PAGE and all sera were examined by immunoblot analysis. For further characterization of the mitochondrial epitopes the antigen fraction was separated in the first dimension according to isoelectric points using isoelectric focusing followed by gel electrophoresis. Proteins recognized by serum antibodies in 2-dimensional immunoblots were analyzed by N-terminal amino acid sequencing. In 1-dimensional immunoblot analysis, 51% of patients with IDC and 34% of controls contained serum antibodies reacting with mitochondrial protein bands with molecular weights of about 30, 43, 60, and preferentially 50 to 55 and 70 to 75 kD (p <0.01). We identified a 75-kD subunit of nicotinamide adenine dinucleotide dehydrogenase (17% in IDC patients vs 5% in controls, p <0.05) and 2 core proteins of ubiquinol-cytochrome-c reductase (core P1: 39% in IDC patients vs 15% in controls, p <0.05; core P2: 20% in IDC patients vs 10% in controls, p <0.1), both enzymes of the respiratory chain, as are most relevant mitochondrial antigens. Furthermore, serum antibodies of patients with IDC were directed against lipoamide-dehydrogenase (15% vs 10% in controls) and a subunit of pyruvate-dehydrogenase (9% vs 3% in controls). Because these antigens play an important role in energy metabolism, the respective antibodies can be more than merely diagnostic markers of cell damage. To attribute them also to pathogenetic relevance appears to be a most attractive but still speculative hypothesis. PMID- 9352976 TI - Morphologic spectrum of primary restrictive cardiomyopathy. AB - A restrictive hemodynamic profile with left ventricular (LV) end-diastolic volume < 100 ml/m2 and LV end-diastolic pressure > 18 mm Hg, in the absence of endomyocardial, pericardial, and specific cardiomyopathy, is a peculiar feature of primary restrictive cardiomyopathy. From 1985 to 1994, 7 hearts of patients who met the above hemodynamic criteria and underwent endomyocardial biopsy because of heart failure, were studied through gross (5 cardiectomies and 2 autopsies), histologic, and electron microscopic investigations. Ages ranged from 9 to 48 years (mean age 29 +/- 13). Four patients (57%) had a positive family history: 2 for hypertrophic and 2 for restrictive cardiomyopathy. Three patterns were identified in the 7 hearts: (1) pure restrictive form in 4 cases with mass/volume ratio 1.2 +/- 0.5 g/ml, ejection fraction 58 +/- 5%, LV end-diastolic volume 67.5 +/- 12.6 ml/m2, LV end-diastolic pressure 26.7 +/- 3.5 mm Hg; (2) hypertrophic-restrictive form in 2 cases with mass/volume ratio 1.5 +/- 0.07 g/ml, ejection fraction 62 +/- 1%, LV end-diastolic volume 69 +/- 10 ml/m2, LV end-diastolic pressure 30 +/- 7 mm Hg; and (3) mildly dilated restrictive form in 1 case with mass/volume ratio 0.9 g/ml, ejection fraction 25%, LV end-diastolic volume 98 ml/m2, LV end-diastolic pressure 40 mm Hg. Histology and electron microscopy disclosed myocardial and myofibrillar disarray and endoperimysial interstitial fibrosis in each pattern. The familial forms suggest the presence of a genetic abnormality. Primary restrictive cardiomyopathy may present with or without hypertrophy and shares similar microscopic pictures with hypertrophic cardiomyopathy. The 2 entities may represent a different phenotypic expression of the same genetic disease. PMID- 9352977 TI - Angiographic and morphologic features of the left ventricle in Ebstein's malformation. AB - Quantitative and qualitative cineangiographic analysis of the left ventricle (LV) was performed in 26 patients with isolated Ebstein's malformation, having a mean age of 23 +/- 17 years. Nine autopsied hearts with isolated Ebstein's malformation were submitted to morphologic and morphometric analysis. In 4 of the cases, it was possible to make a direct correlation between the angiographic data obtained during life and the autopsy findings. On the basis of the LV end diastolic volume we identified 3 groups of patients: 7 with volume <60 ml/m2, another 7 with volume between 60 and 80 ml/m2, and 12 with volume >80 ml/m2. The LV ejection fraction was reduced in 2 patients with normal LV end-diastolic volume and in 6 with increased LV end-diastolic volume. The ratio of ventricular mass to LV end-diastolic volume was always adequate, but a reduction of the ventricular contractive performance (end-systolic pressure to end-systolic volume ratio <3 mm Hg/ml/m2) was found only in patients with a dilated left ventricle. No correlation was demonstrated between the extent of the atrialized component of the right ventricle (mean value 67 +/- 31 cm2, range 13 to 133) and the LV dimensions. All but 2 patients showed a leftward diastolic displacement of the ventricular septum, but in only 1 did this produce an elongated shape of the left ventricle. Sixteen had anomalies of LV dynamics: 10 with hypokinesia (3 of the posterior wall, 4 of the apex, 1 of the inferior wall, 1 of the septum, and 1 global), 6 with dyskinesia (1 of the posterior wall, 2 of the apex, 1 of the posterior wall and apex, 1 of the superior part of the septum, and 1 of the anterior wall), and 8 with premature diastolic distension of the anterobasal wall. Morphometric analysis produced mean values for myocytes of 59 +/- 10%, for the interstitium of 21 +/- 4%, and for fibrous tissue of 20 +/- 9% (normal 4 +/- 1%). Five autopsied hearts had a prolapsing and/or dysplastic mitral valve. PMID- 9352978 TI - Comparison of three-dimensional echocardiographic assessment of volume, mass, and function in children with functionally single left ventricles with two dimensional echocardiography and magnetic resonance imaging. AB - Diminished systolic function or inappropriate hypertrophy are considered risk factors for outcome following the Fontan procedure. These parameters are difficult to assess in univentricular hearts that do not conform to the uniform shapes prescribed by conventional 2-dimensional imaging volume algorithms. Three dimensional echocardiography requires no geometric assumptions and has been validated in both normal and distorted left ventricles. To assess the feasibility and accuracy of this technique in patients with univentricular hearts, we compared 2- and 3-dimensional echocardiographic estimates of ventricular volume, ejection fraction, and mass in patients with functionally single left ventricles with results obtained by magnetic resonance imaging (MRI). Twelve patients with functionally single left ventricles (6 months to 22 years) underwent examination by all 3 modalities. Correlation and agreement with MRI were calculated for volumes, ejection fraction, and mass. Three-dimensional echocardiographic comparison with MRI yielded a bias of 3.4 +/- 5.5 ml and 14.2 +/- 8.3 ml for systolic and diastolic volumes, respectively. Agreement analysis for mass showed a bias of 5.8 +/- 8.4 grams. Two-dimensional echocardiography showed less agreement for both volumes and mass (bias of -2.9 +/- 8.1, 2.9 +/- 10.4 ml and 8.3 +/- 12.0 g for volume and mass, respectively, p >0.05). Ejection fraction by 3-dimensional echocardiography showed significantly closer agreement with MRI (bias of 4.4 +/- 5.3%) than 2-dimensional echocardiography (bias of 8.5 +/- 10.3%, p = 0.04). Thus, 3-dimensional echocardiography provides estimates of ventricular volumes, ejection fraction, and mass that are comparable to MRI in this select group of patients with single ventricles of left ventricular morphology. PMID- 9352979 TI - Frequency of deep vein thrombosis in patients with patent foramen ovale and ischemic stroke or transient ischemic attack. AB - To evaluate the additional value of transesophageal (TEE) compared with transthoracic (TTE) echocardiography and the role of patent foramen ovale (PFO) and deep vein thrombosis in the work-up of embolic events, patients with presumed cardiac embolic stroke or transient ischemic attack (neurovascular etiology was excluded) were prospectively studied by transthoracic and transesophageal contrast echocardiography. If PFO was detected echocardiographically, PFO size was assessed semiquantitatively and phlebography of both legs was performed. Two hundred forty-two consecutive patients (153 men, 60 +/- 15 years) were studied. In 197 patients, neuroimaging showed evidence of embolic infarction. TEE identified 138 potential cardiac sources of embolism in 111 patients, compared with 69 by TTE (p <0.01) in 59 patients. TEE detected potential cardiac sources in 52 patients with negative TTE examination and was significantly superior compared with TTE for identifying left atrial thrombi, spontaneous echo contrast, PFO, atrial septal aneurysm, and atheroma of the ascending aorta. In patients with a positive TTE, additional diagnostic information by TEE was found in only 6 patients and did not change therapy. Phlebography was performed in 53 patients with PFO and revealed deep vein thrombosis in 5 patients (9.5%); all had medium or large PFOs. Thus, in patients with cerebral ischemia of suspected cardiogenic origin and a normal TTE examination, TEE detects potential causes of embolism in 31% of patients and is therefore of diagnostic relevance. Conversely, in the presence of a diagnostic TTE an additional TEE confers only marginal diagnostic benefit. Deep venous thrombosis was detected in nearly 10% of patients with PFO as the sole identifiable cardiac risk factor. Given that in 4 of 5 patients deep vein thrombosis was clinically silent, phlebography should be performed in patients with medium or large interatrial shunts if paradoxical embolism is suspected. PMID- 9352980 TI - Role of nitric oxide in the vasodilator response to mental stress in normal subjects. AB - Vascular production of nitric oxide (NO) plays an important role in a variety of physiologic processes. This study examines the contribution of NO to the vasodilator response to mental stress. The effects of mental arithmetic testing on forearm vascular dynamics were analyzed in 15 normal subjects (9 men; age 45 +/- 12 years) during intraarterial infusion of either saline or N(G)-monomethyl-L arginine (L-NMMA; 4 micromol/min for 15 minutes), an inhibitor of NO synthesis. The effect of L-NMMA on endothelium-independent vasodilation induced by intraarterial infusion of sodium nitroprusside was also studied in 11 of the 15 subjects. Forearm blood flow was measured by plethysmography. Mental stress increased forearm blood flow from 2.35 +/- 0.84 to 5.06 +/- 2.66 ml/min/dl (115%) during saline and from 1.72 +/- 0.59 to 2.81 +/- 0.99 ml/min/dl (63%) during L NMMA infusion. The vasodilator effect of mental stress was significantly lower during L-NMMA infusion than during saline (1.1 +/- 0.65 vs 2.71 +/- 2.15 ml/min/dl; p = 0.01). L-NMMA administration did not significantly change mean arterial pressure and heart rate responses to mental stress. In contrast, the vasodilator effect of sodium nitroprusside (1.6 microg/min) was similar during infusion of L-NMMA and during saline (3.75 +/- 1.55 vs 2.85 +/- 1.38 ml/min/dl; p = 0.16). These findings indicate that local release of NO is involved in the forearm vasodilator response to mental stress. PMID- 9352981 TI - Comparison of peak serum C-reactive protein and hydroxybutyrate dehydrogenase levels in patients with acute myocardial infarction treated with alteplase and streptokinase. AB - Peak serum C-reactive protein concentrations were measured in 146 patients randomized to receive streptokinase, alteplase, or a combination of streptokinase and alteplase in the GUSTO-I trial. Those receiving alteplase treatment had lower values than those receiving streptokinase or the combination treatment. Irrespective of treatment, complete reperfusion of the infarct-related artery (TIMI grade 3 flow) was associated with low peak serum C-reactive protein values. PMID- 9352982 TI - Clinical and angiographic implications of balloon rupture during coronary stenting. AB - Balloon rupture was detected in 66 consecutive patients (5.8%) during coronary stenting. This rare phenomenon usually does not have clinical or angiographic sequelae, but in some cases, it may induce new coronary dissections that can be managed with additional stenting, but also may cause clinical complications. PMID- 9352983 TI - Relation of absence of ST reelevation immediately after reperfusion and success of reperfusion with myocardial salvage. AB - To examine whether resolution in ST elevation without ST reelevation immediately after reperfusion indicates successful reperfusion with myocardial salvage, we studied 40 patients who had an extensive acute myocardial infarction with early reperfusion: 24 patients had ST reelevation and 16 patients had no ST reelevation. Results indicate that (1) in the group with ST reelevation, rapid progression of myocardial damage occurs by reperfusion itself (i.e., reperfusion injury) and (2) in the group without ST reelevation, myocardial damage had already been extensive and irreversible at the time of reperfusion; thus, the absence of ST reelevation is not always a sign of reperfusion with myocardial salvage. PMID- 9352984 TI - Early ambulation after coronary angioplasty and stenting with six French guiding catheters and low-dose heparin. AB - The safety and feasibility of early ambulation was evaluated prospectively in 907 patients undergoing elective coronary angioplasty and stenting with the use of 6Fr guiding catheters, low-dose heparin (5,000 IU), and immediate postprocedural sheath removal by comparing ambulation after 4 hours with immobilization for at least 12 hours. Because no excess in puncture site complications (2.3% vs 2.2%) could be demonstrated after 4-hour ambulation, it is concluded that early ambulation after 6Fr guiding catheter angioplasty by the femoral route with low dose heparin is feasible, safe, and may facilitate a shorter hospital stay. PMID- 9352985 TI - Repeat coronary angiography in patients with chest pain and previously normal coronary angiogram. AB - Patients with chest pain and normal coronary angiograms have excellent long-term survival and are unlikely over the ensuing years to develop clinically significant atherosclerotic coronary artery disease. Specifically, of the 17 subjects with a previously normal coronary angiogram who had repeat angiography an average of almost 9 years later, 15 showed no appearance of coronary artery disease and 2 developed single-vessel coronary artery disease, 1 of whom had a myocardial infarction. PMID- 9352987 TI - Clinical improvement after atrioventricular nodal ablation for atrial fibrillation does not correlate with improved ejection fraction. AB - A retrospective review of 15 patients with atrial fibrillation and class III to IV congestive heart failure who underwent atrioventricular nodal ablation demonstrated a marked improvement in their functional abilities. This improvement, however, could not be explained by the improvement in ejection fraction alone. PMID- 9352986 TI - Rationale, design, and baseline characteristics of the Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial (PREVENT). AB - The Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial is the first angiographic clinical trial to be designed to test whether an agent can slow or even reverse the progression of early coronary atherosclerosis in patients with documented disease. In addition, a subset of patients are undergoing carotid ultrasound examinations, providing a unique opportunity to assess and correlate disease progression in 2 arterial beds. PMID- 9352988 TI - Effects of chronic vasodilator therapy to enhance susceptibility to vasovagal syncope during upright tilt testing. AB - To evaluate the effect of chronic vasodilator therapy on susceptibility to vasovagal syncope, 45 patients with syncope and a positive response to tilt testing were randomly assigned to continue or to discontinue vasodilators. The study result demonstrated that chronic vasodilator therapy enhances susceptibility to vasovagal reaction during upright tilt testing. PMID- 9352989 TI - Accuracy of an automatic and patient-triggered long-term solid memory ambulatory cardiac event recorder. AB - This study evaluated the R-Test Evolution, a new type of cardiac event recorder bearing both patient-triggered and automatic capabilities. Its 7-day automatic arrhythmia analysis showed promising clinical advantages, especially when investigating patients with unexplained rare events such as syncope, feeling of weakness or faintness, palpitations, stroke, or in patients inconsistent in their use of patient-triggered recordings. PMID- 9352990 TI - Effect of biphasic waveforms on transvenous defibrillation thresholds in patients with coronary artery disease. AB - This study is a prospective, randomized comparison of monophasic and biphasic defibrillation thresholds in 19 patients with a single transvenous lead. Despite using reverse polarity and optimal tilts for the monophasic waveform, the defibrillation threshold was reduced with biphasic shocks from 15.8 +/- 11.3 to 11.5 +/- 6.1 (p <0.05) with comparable reductions of leading edge voltage and current. PMID- 9352991 TI - Long-term carvedilol therapy increases parasympathetic nervous system activity in chronic congestive heart failure. AB - To determine the effect of beta blockade on parasympathetic nervous system activity, we assessed RR variability during 24-hour Holter monitoring in 10 patients with congestive heart failure before and after 3 to 4 months of treatment with the beta blocker carvedilol. High-frequency power increased from 26 to 64 ms2, root-mean-square of successive differences in RR interval increased from 14.3 to 23.7 ms2, and percentage of absolute differences >50 ms between successive normal RR intervals increased from 0.8% to 4.7%, all p <0.01, indicating a substantial increase in parasympathetic modulation of RR intervals. PMID- 9352992 TI - Echocardiographic assessment of cardiac involvement in systemic AL amyloidosis in relation to whole body amyloid load measured by serum amyloid P component (SAP) clearance. AB - The severity of cardiac infiltration in AL amyloidosis is unrelated to whole body amyloid load as measured by serum amyloid P (SAP) tracer studies. Radiolabeled SAP and echocardiography permit identification of patients with severe cardiac disease with a low whole body load who may be the best candidates for transplantation. PMID- 9352993 TI - Usefulness of transesophageal imaging of flow convergence region in the operating room for evaluating isolated patent ductus arteriosus. AB - Transesophageal echocardiography (TEE) was performed in 21 patients with isolated patent ductus arteriosus (PDA) with a color Doppler flow convergence method during surgical closure of the ductus. Evaluation of PDA by TEE with the flow convergence method may provide valuable information during surgery and/or thorascopic ductus clipping. PMID- 9352995 TI - Floating in fat: fat kids and fat adults. PMID- 9352994 TI - Acute pulmonary toxicity in an infant from intravenous amiodarone. AB - Intravenous amiodarone is an effective treatment for supraventricular and ventricular tachyarrhythmias. We report a case of acute pulmonary toxicity in an infant from intravenous amiodarone and describe the clinical evaluation and laboratory studies leading to the diagnosis. PMID- 9352996 TI - A misplaced decimal of digitalis dose and tiny Jose Eric Martinez dies. PMID- 9352997 TI - Enalapril, aspirin interaction. PMID- 9352998 TI - What is a stent and where can you get one? PMID- 9352999 TI - The inverse Nehb J lead. PMID- 9353000 TI - Quantification of vascular collagen in atherectomy specimens. PMID- 9353008 TI - Interleukin-12 is indispensable for protective immunity against Leishmania major. AB - Interleukin-12 (IL-12)-deficient mice derived from a strain genetically resistant to infection with Leishmania major were recently shown to be susceptible toward this parasite, developing a strong Th2 response after injection of a large number of parasites. We further investigated the role of IL-12 in L. major infection by studying the responses of mutant mice against smaller numbers of parasites. IL-12 deficient mice infected with only small numbers of parasites showed the progressive lesion development and high parasite burden associated with a polarized Th2 response. Our data show that IL-12 is indispensable for protective immunity against L. major. Even at low inocula, no salvage pathway appears to compensate for the lack of IL-12. However, genetically susceptible BALB/c mice infected with small numbers of parasites were able to resolve lesions and restrict the parasite burden to levels which were 10(5)-fold lower than those in IL-12-deficient mice. In contrast to mutant mice, BALB/c mice mounted a type 1 response against low inocula of L. major. IL-12-deficient BALB/c mice, however, developed a type 2 response. These data emphasize the essential role of IL-12 in resistance against L. major. In addition, this study suggests that in the absence of IL-12, susceptibility to L. major is determined by the inability to induce a Th1 response rather than the development of a Th2 response. Our results are relevant for potential vaccination strategies that use low inocula of infective microorganisms which fail to induce a protective type 1 response at higher inocula. PMID- 9353007 TI - A protective epitope of Moraxella catarrhalis is encoded by two different genes. AB - The high-molecular-weight UspA protein of Moraxella catarrhalis has been described as being both present on the surface of all M. catarrhalis disease isolates examined to date and a target for a monoclonal antibody (MAb 17C7) which enhanced pulmonary clearance of this organism in a mouse model system (M. E. Helminen et al., J. Infect. Dis. 170:867-872, 1994). A recombinant bacteriophage that formed plaques which bound MAb 17C7 was shown to contain a M. catarrhalis gene, designated uspA1, that encoded a protein with a calculated molecular weight of 88,271. Characterization of an isogenic uspA1 mutant revealed that elimination of expression of UspA1 did not eliminate the reactivity of M. catarrhalis with MAb 17C7. In addition, N-terminal amino acid analysis of internal peptides derived from native UspA protein and Southern blot analysis of M. catarrhalis chromosomal DNA suggested the existence of a second UspA-like protein. A combination of epitope mapping and ligation-based PCR methods identified a second M. catarrhalis gene, designated uspA2, which also encoded the MAb 17C7-reactive epitope. The UspA2 protein had a calculated molecular weight of 62,483. Both the isogenic uspA1 mutant and an isogenic uspA2 mutant possessed the ability to express a very-high-molecular-weight antigen that bound MAb 17C7. Southern blot analysis indicated that disease isolates of M. catarrhalis likely possess both uspA1 and uspA2 genes. Both UspA1 and UspA2 most closely resembled adhesins produced by other bacterial pathogens. PMID- 9353009 TI - Borrelia burgdorferi induces chemokines in human monocytes. AB - Lyme disease is clinically and histologically characterized by strong inflammatory reactions that contrast the paucity of spirochetes at lesional sites, indicating that borreliae induce mechanisms that amplify the inflammatory response. To reveal the underlying mechanisms of chemoattraction and activation of responding leukocytes, we investigated the induction of chemokines in human monocytes exposed to Borrelia burgdorferi by a dose-response and kinetic analysis. Lipopolysaccharide (LPS) derived from Escherichia coli was used as a positive control stimulus. The release of the CXC chemokines interleukin-8 (IL-8) and GRO-alpha and the CC chemokines MIP-1alpha, MCP-1, and RANTES was determined by specific enzyme-linked immunosorbent assays, and the corresponding gene expression patterns were determined by Northern blot analysis. The results showed a rapid and strong borrelia-inducible gene expression which was followed by the release of chemokines with peak levels after 12 to 16 h. Spirochetes and LPS were comparably effective in stimulating IL-8, GRO-alpha, MCP-1, and RANTES expression, whereas MIP-1alpha production preceded and exceeded chemokine levels induced by LPS. Unlike other bacteria, the spirochetes themselves did not bear or release factors with intrinsic chemotactic activity for monocytes or neutrophils. Thus, B. burgdorferi appears to be a strong inducer of chemokines which may, by the attraction and activation of phagocytic leukocytes, significantly contribute to inflammation and tissue damage observed in Lyme disease. PMID- 9353010 TI - Genetic and biochemical analyses of Actinobacillus pleuropneumoniae urease. AB - The urease gene cluster from the virulent Actinobacillus pleuropneumoniae serotype 1 strain CM5 was cloned and sequenced. The urease activity was associated with a 6.3-kbp region which contains eight long open reading frames (ORFs). The structural genes, ureABC, are separated from the accessory genes, ureEFGD, by a 615-bp ORF of unknown function, ureX. Homologies were found with the structural and accessory urease gene products of Haemophilus influenzae and, to a lesser extent, with those of other organisms. The urease enzyme subunits had predicted molecular masses of 61.0, 11.3, and 11.0 kDa, and the size of the holoenzyme was estimated to be 337 +/- 13 kDa by gel filtration chromatography. Urease activity was maximal but unstable at 65 degrees C. In cell lysates, the A. pleuropneumoniae urease was stable over a broad pH range (5.0 to 10.6) and the optimal pH for activity was 7.7. The Km was 1.5 +/- 0.1 mM urea when it was assayed at pH 7.7. The low Km suggests that this enzyme would be active in the respiratory tract environment, where urea levels should be similar to those normally found in pig serum (2 to 7 mM). PMID- 9353011 TI - Role of lipopolysaccharide in signaling to subepithelial polymorphonuclear leukocytes. AB - Polymorphonuclear leukocyte (PMN) infiltration and migration across colonic intestinal epithelia is a hallmark of inflammation in Shigella flexneri-mediated dysentery. To identify bacterial signals associated with this process, potential stimulatory factors mediating initial PMN association with the epithelium and subsequent transepithelial migration were examined in an in vitro model system. Quantitative analyses revealed that purified S. flexneri lipopolysaccharide (LPS) deposited at the apical surface of polarized intestinal epithelial cells transcytosed to the basolateral pole, a process dependent on the stage of epithelial cell differentiation. Transcytosed LPS in the presence of normal human serum (NHS), a source of LPS binding protein and soluble CD14, mediated both interleukin-8 secretion at the basolateral pole and enhanced PMN adherence. In addition, LPS stimulated a significant degree of directed transepithelial migration of PMNs, an event that was further enhanced in the presence of NHS. These results implicate LPS in signaling subepithelial PMN emigration and enhancing PMN-epithelium interactions prior to and during subsequent Shigella induced transepithelial migration. PMID- 9353012 TI - Interleukin-12 gene expression in human monocyte-derived macrophages stimulated with Mycobacterium bovis BCG: cytokine regulation and effect of NK cells. AB - Macrophage-derived interleukin-12 (IL-12) is essential for the activation of a protective immune response against intracellular pathogens. In this study, we examined the regulation of IL-12 mRNA expression by monocyte-derived macrophages (MDM) in response to Mycobacterium bovis BCG stimulation. A reverse transcription PCR assay detected p40 mRNA of IL-12 at 3 h and showed a peak at 6 to 12 h with a subsequent decline. Semiquantitation of mRNA levels by competitive PCR revealed that pretreatment with gamma interferon (IFN-gamma) amplified the expression approximately 100-fold, while pretreatment with tumor necrosis factor alpha (TNF alpha) or granulocyte-macrophage colony-stimulating factor augmented this expression about 10-fold. In contrast, pretreatment with IL-10 and IL-4 inhibited IL-12 mRNA expression. These results were further confirmed by measuring the p70 bioactive protein level in each conditioned medium by an enzyme-linked immunosorbent assay. Since IL-12 mRNA expression was weak without cytokine pretreatment and IFN-gamma strongly augmented production, we speculated that IFN gamma might have a role in BCG stimulation of IL-12 mRNA expression. Unexpectedly, the addition of three different kinds of anti-IFN-gamma antibodies and anti-IFN-gamma receptor antibody and the coaddition of anti-TNF-alpha antibody with anti-IFN-gamma receptor antibody all failed to inhibit IL-12 mRNA expression. However, the MiniMACS method used to remove NK cells from a mononuclear cell suspension inhibited the expression of p40 mRNA but not the expression of mRNA of TNF-alpha or IL-1beta. We concluded that the coexistence of NK cells was essential for the induction of IL-12 in MDM stimulated with BCG rather than through the secretion of IFN-gamma. PMID- 9353013 TI - A role for pneumolysin but not neuraminidase in the hearing loss and cochlear damage induced by experimental pneumococcal meningitis in guinea pigs. AB - We investigated the roles of pneumolysin and neuraminidase in the pathogenesis of deafness and cochlear damage during experimental pneumococcal meningitis. Anesthetized guinea pigs were inoculated intracranially with 7.5 log10 CFU of either (i) wild-type Streptococcus pneumoniae D39 (n = 8), (ii) PLN-A, a defined isogenic derivative of D39 deficient in pneumolysin (n = 5), or (iii) deltaNA1, a new derivative of D39 deficient in neuraminidase constructed by insertion duplication mutagenesis of the nanA gene (n = 5). To quantify hearing loss, the auditory nerve compound action potential evoked by a tone pulse was recorded from the round window membrane of the cochlea every 3 h for 12 h. The organ of Corti was intravitally fixed for subsequent examination by high-resolution scanning and transmission electron microscopy. All animals sustained similar meningeal inflammatory responses. PLN-A induced significantly less hearing loss than D39 over the frequency range of 3 to 10 kHz. Levels of mean hearing loss at 10 kHz 12 h postinoculation were as follows: D39, 50 dB; deltaNA1, 52 dB (P = 0.76 versus D39), and PLN-A, 12 dB (P < 0.0001 versus D39). The mean rates of hearing loss at 10 kHz were 4.4 dB/h for D39, 4.3 dB/h for deltaNA1, and just 1.0 dB/h for PLN-A (P < 0.0001 versus D39). Suppurative labyrinthitis was universal. PLN-A induced the accumulation of less protein in the cerebrospinal fluid (P = 0.04 versus D39). Infection with D39 and deltaNA1 induced significant damage to the reticular lamina, the sensory hair cells, and supporting cells of the organ of Corti. By contrast, after infection with PLN-A, the organ of Corti appeared virtually intact. Pneumolysin seems to be the principal cause of cochlear damage in this model of meningogenic deafness. No clear pathogenic role was demonstrated for neuraminidase. PMID- 9353015 TI - Immunogenicity and protective immunity induced by synthetic peptides associated with a catalytic subdomain of mutans group streptococcal glucosyltransferase. AB - We examined the immunogenicity and induction of protective immunity of two 19-mer sequences (GGY and AND) which overlapped a highly conserved region which has recently been implicated in the enzymatic activity of glucosyltransferases (GTFs) of the mutans group streptococci. These peptides were synthesized as eight branched constructs on a lysine core. Serum immunoglobulin G (IgG) antibody, induced by subcutaneous (s.c. [salivary gland vicinity]) injection with these peptide constructs, reacted with the inciting antigen, with mutans streptococcal GTFs, and with a 21-mer peptide (CAT) containing an aspartate previously shown to covalently bind sucrose. Several of these antisera also inhibited the ability of Streptococcus sobrinus GTF to synthesize insoluble glucan. Significant levels of salivary IgA antibody were also induced by GGY and AND peptide constructs after s.c. injection. The effect of immunization with the GGY and AND peptide constructs on the cariogenicity of Streptococcus mutans was studied in three experiments by immunization of weanling Sprague-Dawley rats, twice at 7- to 14 day intervals with peptides, S. sobrinus GTF, or phosphate-buffered saline. All rats were then orally infected with S. mutans SJ. After 63-day infection periods, the GGY and AND-injected groups had significant dental caries reductions compared with sham-injected groups in most experiments. These studies support the existence of an additional catalytic subdomain within the sequence defined by the GGY and AND peptides. Furthermore, the epitopes defined in these sequences have significant immunogenicity, can induce immune responses which interfere with GTF mediated glucan synthesis in vitro, and can protect rats from experimental dental caries. PMID- 9353014 TI - Immunization with a recombinant C-terminal fragment of Plasmodium yoelii merozoite surface protein 1 protects mice against homologous but not heterologous P. yoelii sporozoite challenge. AB - It has been reported previously that immunization with recombinant protein containing the two epidermal growth factor (EGF)-like modules from merozoite surface protein 1 (MSP-1) of Plasmodium yoelii (strain YM) protects mice against a lethal blood-stage challenge with the same parasite strain. Since MSP-1 is expressed in both liver- and blood-stage schizonts and on the surface of merozoites, we evaluated the effectiveness of immunization with recombinant proteins containing either the individual or the two combined EGF-like modules in producing a protective response against a sporozoite challenge. The recombinant protein expressing the combined EGF-like modules of the YM strain protected mice against a homologous sporozoite challenge, and sterile protection, as defined by the absence of detectable blood-stage parasites, was observed in the majority of the mice. In contrast, mice immunized with recombinant P. yoelii YM MSP-1 were not protected against a heterologous challenge with sporozoites from strain 265 BY of P. yoelii. The lack of protection may be explained by differences identified in the amino acid sequences of MSP-1 for the two strains. A recombinant protein containing the two EGF-like modules of MSP-1 from P. yoelii 265 BY was produced and used to immunize mice. These mice were protected against a homologous challenge with sporozoites of P. yoelii 265 BY. The results suggest that a recombinant MSP-1 has potential as a vaccine against malaria, but its efficacy may be limited by sequence polymorphism and selection of variants. PMID- 9353016 TI - Evaluation of the virulence of nontypeable Haemophilus influenzae lipooligosaccharide htrB and rfaD mutants in the chinchilla model of otitis media. AB - Considerable evidence has implicated nontypeable Haemophilus influenzae (NTHi) lipooligosaccharide (LOS) in the pathogenesis of otitis media (OM); however, its exact role has not been conclusively established. Recently, two NTHi LOS deficient mutants have been created and described. Strain 2019-DK1, an rfaD gene mutant, expresses a truncated LOS consisting of only three deoxy-D-manno octulosonic acid residues, a single heptose, and lipid A. Strain 2019-B29, an isogenic htrB mutant, possesses an altered oligosaccharide core and an altered lipid A. Each strain's ability to colonize the nasopharynx and to induce OM subsequent to transbullar inoculation was evaluated in the chinchilla model. Nasopharyngeal colonization data indicate that the parent strain and both mutants are able to colonize the nasopharynx and exhibit comparable clearance kinetics. Compared with the parent and each other, however, the mutants demonstrated marked differences in virulence regarding their relative abilities to induce OM and persist in the middle ear post-transbullar inoculation. Strain B29 required a 3 log-greater dose to induce OM than the parent strain and did not exhibit evidence of sustained multiplication but persisted for the same duration as the parent. Conversely, strain-DK1, even when inoculated at a dose 4 logs greater than the parent dose, was eliminated from the middle ear 72 h after challenge. A comparison of the relative pathogenicities of these isolates provides the opportunity to address fundamental questions regarding the contribution of LOS to pathogenesis issues at the molecular level. Specifically, the impact of these LOS gene disruptions on OM pathogenesis can be defined and may thus provide potential new targets for future protection and intervention strategies. PMID- 9353017 TI - Sialylation of Neisseria meningitidis lipooligosaccharide inhibits serum bactericidal activity by masking lacto-N-neotetraose. AB - Exogenous sialylation of gonococcal lipooligosaccharide causes resistance to serum bactericidal activity. The aim of this study was to determine how lipooligosaccharide sialylation affects the serum sensitivities of group C Neisseria meningitidis strains. The relationship between the degree of sialylation or expression of the lipooligosaccharide sialic acid acceptor, lacto N-neotetraose (LNnT), of nine meningococcal strains and their sensitivities to a pool of normal human sera was assessed. All strains expressed LNnT that was variously endogenously sialylated. Susceptibility to serum bactericidal activity ranged from extremely sensitive to resistant in 50% serum. For endogenously sialylated strains, the amount of killing correlated with the amount of free LNnT above a threshold of expression; strains that expressed less than the threshold survived in 25% serum. All strains added more sialic acid when they were grown in medium that contained cytidine monophospho-N-acetylneuraminic acid. Exogenous sialylation reduced the expression of free LNnT and significantly increased serum resistance. Exogenous sialylation affected killing through both classical and alternative complement pathways. The killing of exogenously sialylated strains also correlated with the amount of free LNnT. The amounts of endogenous, exogenous, and total sialic acid bound to LNnT did not correlate with the resistance of strains to serum bactericidal activity; rather, the loss of free LNnT expression by sialylation was associated with resistance. In conclusion, the expression of free LNnT by group C meningococcal strains is directly associated with the amount of killing of organisms in pooled human sera. Both endogenous and exogenous lipooligosaccharide sialylation are associated with increased serum resistance by masking LNnT. PMID- 9353018 TI - N-glycosylated proteins are involved in efficient internalization of Klebsiella pneumoniae by cultured human epithelial cells. AB - Klebsiella pneumoniae obtained from patients with urinary tract infections is able to invade cultured human epithelial cells. The internalization process is dependent upon both microfilaments and microtubules. To better understand the interaction of these invasive bacteria with the host cell receptor(s), bladder, lung, and ileocecal epithelial cells were infected with K. pneumoniae in the presence of various lectins possessing multiple glycan specificities. It was found that the N-acetylglucosamine (GlcNAc)-specific lectins concanavalin A, Datura stramonium agglutinin, and wheat germ agglutinin significantly inhibited the invasion of K. pneumoniae into these cells but did not interfere with the internalization of an invasive strain of Salmonella typhimurium. Conversely, internalization of K. pneumoniae but not S. typhimurium was also significantly inhibited when the bacteria were pretreated with GlcNAc or chitin hydrolysate, a GlcNAc polymer, prior to the gentamicin invasion assay. Other carbohydrates such as glucose, galactose, mannose, fucose, and N-acetylneuraminic acid had no inhibitory effects on K. pneumoniae uptake. Furthermore, internalization of K. pneumoniae but not S. typhimurium by HCT8 cells was also significantly inhibited when eukaryotic protein glycosylation was interrupted by tunicamycin or when host N-linked surface glycans were removed by pretreatment with N-glycosidase F. These studies suggest that a N-glycosylated protein receptor is involved in the internalization of K. pneumoniae by human epithelial cells in vitro. The results also indicate that internal GlcNAc residues might be a carbohydrate component of the receptor. PMID- 9353019 TI - Comparative analysis of immunoglobulin A1 protease activity among bacteria representing different genera, species, and strains. AB - Immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region are produced constitutively by a number of pathogens, including Haemophilus influenzae, Neisseria meningitidis, Neisseria gonorrhoeae, and Streptococcus pneumoniae, as well as by some members of the resident oropharyngeal flora. Whereas IgA1 proteases have been shown to interfere with the functions of IgA antibodies in vitro, the exact role of these enzymes in the relationship of bacteria to a human host capable of responding with enzyme-neutralizing antibodies is not clear. Conceivably, the role of IgA1 proteases may depend on the quantity of IgA1 protease generated as well as on the balance between secreted and cell-associated forms of the enzyme. Therefore, we have compared levels of IgA1 protease activity in cultures of 38 bacterial strains representing different genera and species as well as strains of different pathogenic potential. Wide variation in activity generation rate was found overall and within some species. High activity was not an exclusive property of bacteria with documented pathogenicity. Almost all activity of H. influenzae, N. meningitidis, and N. gonorrhoeae strains was present in the supernatant. In contrast, large proportions of the activity in Streptococcus, Prevotella, and Capnocytophaga species was cell associated at early stationary phase, suggesting that the enzyme may play the role of a surface antigen. Partial release of cell-associated activity occurred during stationary phase. Within some taxa, the degree of activity variation correlated with degree of antigenic diversity of the enzyme as determined previously. This finding may indicate that the variation observed is of biological significance. PMID- 9353020 TI - A second merozoite surface protein (MSP-4) of Plasmodium falciparum that contains an epidermal growth factor-like domain. AB - Merozoite surface proteins of Plasmodium falciparum play a critical role in the invasion of human erythrocytes by the malaria parasite. Here we describe the identification of a novel protein with a molecular mass of 40 kDa that is found on the merozoite surface of P. falciparum. We call this protein merozoite surface protein 4 (MSP-4). Evidence for the surface location of MSP-4 includes (i) a staining pattern that is consistent with merozoite surface location in indirect immunofluorescent studies of cultured parasites, (ii) localization of MSP-4 in the detergent phase in Triton X-114 partitioning studies, and (iii) nucleotide sequencing studies which predict the presence of an N-terminal signal sequence and a hydrophobic C-terminal sequence in the protein. Immunoprecipitation studies of biosynthetically labelled parasites with [3H] myristic acid indicated that MSP 4 is anchored on the merozoite surface by a glycosylphosphatidylinositol moiety. Of considerable interest is the presence of a single epidermal growth factor-like domain at the C terminus of the MSP-4 protein, making it the second protein with such a structure to be found on the merozoite surface. PMID- 9353021 TI - Misexpression of the white-phase-specific gene WH11 in the opaque phase of Candida albicans affects switching and virulence. AB - Candida albicans WO-1 switches between a white- and an opaque-colony-forming phenotype. The gene WH11 is expressed differentially in the white phase. The WH11 open reading frame was inserted downstream of the promoter of the opaque-phase specific gene OP4 in the transforming vector pCWOP16, and resulting transformants were demonstrated to misexpress WH11 in the opaque phase. Misexpression had no effect on the ability to switch from the white to the opaque or the opaque to the white phase, and it had no effect on the genesis of the unique opaque-phase cellular phenotype, even though the Wh11 protein was distributed throughout the cytoplasm in a manner similar to that observed for the endogenous gene product in the white phase. Misexpression did, however, increase the frequency of the opaque to-white transition 330-fold and markedly increased the virulence of cells in the opaque phase in a mouse tail injection model. PMID- 9353022 TI - Regions of Yersinia pestis V antigen that contribute to protection against plague identified by passive and active immunization. AB - V antigen of Yersinia pestis is a multifunctional protein that has been implicated as a protective antigen, a virulence factor, and a regulatory protein. A series of V-antigen truncates expressed as glutathione S-transferase (GST) fusion proteins (GST-V truncates) have been cloned and purified to support immunogenicity and functionality studies of V antigen. Immunization studies with GST-V truncates have identified two regions of V antigen that confer protection against Y. pestis 9B (a fully virulent human pneumonic plague isolate) in a mouse model for plague. A minor protective region is located from amino acids 2 to 135 (region I), and a major protective region is found between amino acids 135 and 275 (region II). In addition, analysis of IgG titers following immunization suggested that the major antigenic region of V antigen is located between amino acids 135 and 245. A panel of monoclonal antibodies raised against recombinant V antigen was characterized by Western blotting against GST-V truncates, and epitopes of most of the monoclonal antibodies were mapped to region I or II. Monoclonal antibody 7.3, which recognizes an epitope in region II, passively protected mice against challenge with 12 median lethal doses of Y. pestis GB, indicating that region II encodes a protective epitope. This is the first report of a V-antigen-specific monoclonal antibody that will protect mice against a fully virulent strain of Y. pestis. The combined approach of passive and active immunization has therefore confirmed the importance of the central region of the protein for protection and also identified a previously unknown protective region at the N terminus of V antigen. PMID- 9353023 TI - In vitro modulation of proliferation and cytokine production by human peripheral blood mononuclear cells from subjects with various forms of coccidioidomycosis. AB - Using peripheral blood mononuclear cells (PBMC) from individuals with or without coccidioidal delayed-type hypersensitivity (DTH), we examined and attempted to modulate the in vitro responses of PBMC from various donors to the coccidioidal antigen toluene spherule lysate (TSL). Among healthy DTH-positive donors, 100 ng of human recombinant interleukin-10 (IL-10) per ml suppressed both PBMC proliferation (P = 0.01) and gamma interferon (IFN-gamma) and IL-12 production (for both, P < 0.05). In vitro proliferation and production of IFN-gamma and IL 12 by PBMC were significantly higher in DTH-positive donors with active coccidioidomycosis than in healthy, nonimmune controls (P < 0.05) but not in active DTH-negative donors with or without human immunodeficiency virus infection (for both, P > 0.05). Human recombinant IL-12 increased IFN-gamma production by PBMC from active, DTH-positive donors (P = 0.01) but not by PBMC from DTH negative groups. For healthy DTH-positive donors, the median antigen-reactive cell frequency per 10(5) PBMC was 3.7, compared to 1.7 in DTH-negative donors with active coccidioidomycosis (P = 0.03). These data indicate that the in vitro TSL response is highly dependent on coccidioidal DTH. Not only do PBMC from individuals with DTH appear to respond to TSL, but their response can be modulated in vitro with either IL-10 or IL-12. On the other hand, PBMC from DTH negative individuals do not respond in vitro to TSL and their response is not modulable, suggesting a lack of antigen response. PMID- 9353024 TI - Detoxified lipooligosaccharide from nontypeable Haemophilus influenzae conjugated to proteins confers protection against otitis media in chinchillas. AB - Detoxified-lipooligosaccharide (dLOS)-protein conjugates from nontypeable Haemophilus influenzae (NTHi) elicited a significant rise of anti-LOS antibodies with bactericidal activity in rabbits (X.-X. Gu, C.-M. Tsai, T. Ueyama, S. J. Barenkamp, J. B. Robbins, and D. J. Lim, Infect. Immun. 64:4047-4053, 1996). In this study, we evaluated whether vaccination with the conjugates would protect against NTHi otitis media in chinchillas. Fifty-eight chinchillas received three subcutaneous or intramuscular injections of dLOS-conjugated tetanus toxoid, dLOS conjugated high-molecular-weight proteins from NTHi, or saline (control) in Freund's adjuvant and then were challenged by intrabullar inoculation with 140 CFU of NTHi. All vaccinated animals responded with elevated serum titers of anti LOS antibody, and 49% (19 of 39) demonstrated bactericidal activity against the homologous strain. Otitis media with culture-positive NTHi effusions developed in all 19 controls and 56% (22 of 39) of the vaccinated animals during a period of 21 days (P < 0.001). Bacterial counts of the middle ear effusions were lower in the vaccine groups than in the controls (P < 0.01). The incidences of infection in the unchallenged ear or inner ear were 26 or 28% in the vaccine groups and 53 or 58% in the controls (P < 0.05). The signs of infection observed by otoscopy were less severe in the vaccine groups than in the controls. There was no significant difference between the two vaccine groups. These data indicate that active immunization with LOS-based conjugates reduces the incidence of NTHi induced otitis media. PMID- 9353025 TI - Binding of diarrheagenic Escherichia coli to 32- to 33-kilodalton human intestinal brush border proteins. AB - We have detected human intestinal brush border proteins to which Escherichia coli strains adhere by means of a blotting-nitrocellulose method in which the binding of radiolabeled bacteria to sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated intestinal cell membranes was evaluated. The brush border fraction contained several polypeptides that bound only adherent E. coli strains. The most prominent and consistent of these proteins had apparent molecular masses of 32 to 33 kDa. Additional polypeptides ranging from 50 to 70, from 105 to 130, and from 180 to 200 kDa were also recognized by adherent E. coli strains, although with less intensity (in accordance with the number of bound bacteria to these polypeptides). Independently of the pattern of adherence (localized [LA], diffuse [DA], or aggregative [AggA]) all HEp-2-adhering strains recognized, with different intensities, the 32- to 33-kDa brush border proteins, whereas nonadhesive strains did not. The relative avidity of an LA strain to bind to the 32- to 33-kDa proteins was approximately seven- and sixfold higher than the binding of strains with aggregative and diffuse adherence, respectively. Thus, it is reasonable to think that LA, DA, and AggA strains have a common adhesin that mediates binding to the 32- to 33-kDa bands. Inhibition experiments using HEp-2 cells demonstrated that isolated 32- to 33-kDa proteins or specific antiserum blocked preferentially bacterial adherence of the LA pattern. Delipidization and protein digestion of the human brush borders confirmed that E. coli bound to structures of a proteinaceous nature. Deglycosylation studies and sodium meta-periodate oxidation of the intestinal cell membranes decreased bacterial binding activity significantly, indicating that E. coli bound to carbohydrate moieties in the glycoproteins. These results suggest that binding of E. coli strains, mainly of the LA phenotype, to the 32- to 33-kDa proteins could play a role in colonization through adherence to the intestinal mucosa. PMID- 9353026 TI - Role of outer membrane protein H (OmpH)- and OmpA-specific monoclonal antibodies from hybridoma tumors in protection of mice against Pasteurella multocida. AB - Two major outer membrane proteins of Pasteurella multocida, designated OmpH and OmpA, were characterized and shown to be related to the families of porin and heat-modifiable proteins, respectively. The backpack hybridoma tumor system in BALB/c mice was used to continuously deliver immunoglobulin G2b (IgG2b) monoclonal antibodies (MAbs) specific for OmpH (MAb MT1) and OmpA (MAb MT4.1). MAbs were detected in serum and peritoneal lavage samples of mice bearing hybridoma tumors by an enzyme-linked immunosorbent assay and an immunoblot assay. Highly significant protection was observed in mice bearing MT1 hybridoma tumors against both intraperitoneal and intranasal challenge infections with homologous nontoxigenic P. multocida strains possessing MAb MT1-reacting epitopes, whereas the mice bearing MT4.1 hybridoma tumors were not protected. The numbers of P. multocida organisms in the lungs of mice bearing MT1 hybridoma tumors were significantly less than those in lungs of mice bearing MT4.1 hybridoma tumors at 48 h postchallenge. These results indicate that the OmpH-specific MAb inhibited proliferation of P. multocida in the lungs. MAb MT1 was unable to kill P. multocida in vitro in the presence of complement. However, an enhanced phagocytosis by polymorphonuclear cells (PMNs) was observed in mice bearing MT1 hybridoma tumors. P. multocida induced a more extensive and rapid influx of PMNs into the peritoneal cavity of mice bearing MT1 hybridoma tumors than of mice bearing MT4.1 hybridoma tumors. The results of this study demonstrate for the first time that IgG MAbs against OmpH of P. multocida are involved in the protection of mice against lethal challenge infection by means of opsonization and inhibition of proliferation of P. multocida as a result of increased influx of PMNs into the infection site. PMID- 9353027 TI - Th1 response in Salmonella typhimurium-infected mice with a high or low rate of bacterial clearance. AB - Previous studies have shown that the capacity to clear an attenuated strain of Salmonella typhimurium after the second week of infection varies widely among mouse strains. Bacterial clearance is mediated by CD4+ T cells and is regulated in part by the H-2 complex. The aim of the present study was to compare the patterns of cytokine mRNA expression in the spleens of C57BL/6 (H-2b) and CBA (H 2k) mice, which exhibit a low and a high rate of bacterial clearance, respectively. A transient increase in interleukin-12 (IL-12) mRNA levels was found in both mouse strains. Gamma interferon (IFN-gamma) gene expression was higher and more sustained in C57BL/6 than in CBA mice. No increase in IL-4 mRNA was detected. A transient increase in IL-10 mRNA was found in C57BL/6 mice. Separation of spleen cells into CD4+ and CD4- fractions showed that CD4+ T cells produced the bulk of IFN-gamma in both mouse strains and of IL-10 in C57BL/6 mice. Infection of H-2 congenic mice induced a higher level of IFN-gamma mRNA expression by CD4+ T cells in mice with a low rate of clearance (H-2b) than in mice with a high rate of clearance (H-2q). Treatment of infected C57BL/6 mice with anti-IFN-gamma or anti-CD4 monoclonal antibodies indicated that IFN-gamma participates in resistance in the early phase of infection, but not in bacterial clearance, and that CD4+ T cells mediate bacterial clearance during the 3rd week of infection. Taken together, these results suggest that defective bacterial clearance in H-2b mice is not linked to defective IFN-gamma production and that CD4+ T cells mediate bacterial clearance by an IFN-gamma-independent mechanism. PMID- 9353028 TI - Definition of Mycobacterium tuberculosis culture filtrate proteins by two dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry. AB - A number of the culture filtrate proteins secreted by Mycobacterium tuberculosis are known to contribute to the immunology of tuberculosis and to possess enzymatic activities associated with pathogenicity. However, a complete analysis of the protein composition of this fraction has been lacking. By using two dimensional polyacrylamide gel electrophoresis, detailed maps of the culture filtrate proteins of M. tuberculosis H37Rv were generated. In total, 205 protein spots were observed. The coupling of this electrophoretic technique with Western blot analysis allowed the identification and mapping of 32 proteins. Further molecular characterization of abundant proteins within this fraction was achieved by N-terminal amino acid sequencing and liquid chromatography-mass spectrometry. Eighteen proteins were subjected to N-group analysis; of these, only 10 could be sequenced by Edman degradation. Among the most interesting were a novel 52-kDa protein demonstrating significant homology to an alpha-hydroxysteroid dehydrogenase of Eubacterium sp. strain VPI 12708, a 25-kDa protein corresponding to open reading frame 28 of the M. tuberculosis cosmid MTCY1A11, and a 31-kDa protein exhibiting an amino acid sequence identical to that of antigen 85A and 85B. This latter product migrated with an isoelectric point between those of antigen 85A and 85C but did not react with the antibody specific for this complex, suggesting that there is a fourth member of the antigen 85 complex. Novel N-terminal amino acid sequences were obtained for three additional culture filtrate proteins; however, these did not yield significant homology to known protein sequences. A protein cluster of 85 to 88 kDa, recognized by the monoclonal antibodies IT-57 and IT-42 and known to react with sera from a large proportion of tuberculosis patients, was refractory to N-group analysis. Nevertheless, mass spectrometry of peptides obtained from one member of this complex identified it as the M. tuberculosis KatG catalase/peroxidase. Thus, the detailed mapping of M. tuberculosis proteins, combined with state-of-the-art analytical techniques such as mass spectrometry, provides a basis for further analysis and rapid identification of biologically relevant molecules. PMID- 9353029 TI - Immunization with heat-killed Mycobacterium vaccae stimulates CD8+ cytotoxic T cells specific for macrophages infected with Mycobacterium tuberculosis. AB - Immune responses to Mycobacterium tuberculosis are analyzed in mice which have been immunized with Mycobacterium vaccae to examine novel ways of altering protective immunity against M. tuberculosis. The spleen cells of mice immunized with M. vaccae proliferate and secrete gamma interferon (IFN-gamma) in response to challenge with live M. tuberculosis in vitro. Immunization with M. vaccae results in the generation of CD8+ T cells which kill syngeneic macrophages infected with M. tuberculosis. These effector cytotoxic T cells (CTL) are detectable in the spleen at 2 weeks after immunization with M. vaccae but cannot be found in splenocytes 3 to 6 weeks postimmunization. However, M. tuberculosis specific CTL are revealed following restimulation in vitro with heat-killed M. vaccae or M. tuberculosis, consistent with the activation of memory cells. These CD8+ T cells secrete IFN-gamma and enhance the production of interleukin 12 when cocultured with M. tuberculosis-infected macrophages. It is suggested that CD8+ T cells with a cytokine secretion profile of the Tc1 class may themselves maintain the dominance of a Th1-type cytokine response following immunization with M. vaccae. Heat-killed M. vaccae deserves attention as an alternative to attenuated live mycobacterial vaccines. PMID- 9353030 TI - A novel component different from endotoxin extracted from Prevotella intermedia ATCC 25611 activates lymphoid cells from C3H/HeJ mice and gingival fibroblasts from humans. AB - A novel immunobiologically active fraction was prepared from a phenol-water extract of Prevotella intermedia ATCC 25611 by Sephadex G-100 column chromatography. The fraction consisted mainly of carbohydrate and protein and was devoid of fatty acid. The fraction showed high-molecular-weight bands (10,000 to 12,000) on deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE) and was scarcely active in a Limulus test. We designated the fraction Prevotella glycoprotein (PGP). The PGP fraction showed strong mitogenicity on splenocytes and cytokine-inducing activities on peritoneal macrophages from both C3H/HeJ and C3H/HeN mice, and it stimulated human gingival fibroblasts to produce cytokines. The activities of the PGP fraction were resistant to heat inactivation (100 degrees C for 1 h) and protease treatments and were scarcely inhibited by polymyxin B. In contrast, the purified lipopolysaccharide fraction (LPS-PCP) extracted from the same bacterium with a phenol-chloroform-petroleum ether mixture, which showed strong Limulus activity and a single low-molecular-weight band (approximately 3,000) on DOC-PAGE, lacked the activities on splenocytes and macrophages from C3H/HeJ mice and human gingival fibroblasts. The activities of the LPS-PCP fraction on cells from C3H/HeN mice were completely inhibited by polymyxin B. The LPS extracted from the same bacterium with hot phenol-water (LPS PW) exhibited the properties of both the PGP fraction and the LPS-PCP fraction. These findings suggest that the unique bioactivities of the LPS-PW fraction of oral black-pigmented bacteria reported to date, which differed from those of the classical endotoxin, were derived from the PGP fraction and not from the LPS itself. PMID- 9353031 TI - Response of Chlamydia trachomatis serovar E to iron restriction in vitro and evidence for iron-regulated chlamydial proteins. AB - Iron is a well-established mediator of virulence in several bacterial pathogens, yet little is known about the role of iron in infectious disease processes caused by obligate intracellular bacterial pathogens. In this study, the effect of iron limitation was examined for the sexually transmitted infectious agent Chlamydia trachomatis in an in vitro model of human genital infection using the intracellular iron-chelating reagent deferoxamine mesylate (Desferal). Iron restriction caused a significant reduction in infectivity of C. trachomatis elementary bodies (EB) harvested from Desferal-exposed polarized epithelial cells when compared to that of EB harvested from iron-sufficient control cell cultures. Replacement of the Desferal exposure medium with medium containing iron-saturated transferrin restored chlamydial infectivity, whereas replacement with growth medium alone had no effect. The following three prominent morphological features were observed by electron microscopic examination of chlamydia-infected cells exposed to Desferal: (i) inclusions containing chlamydiae greatly delayed in maturation, (ii) substantial blebbing within chlamydial inclusions, and (iii) electron-dense material surrounding inclusions. Protein analyses of highly purified EB by two-dimensional polyacrylamide gel electrophoresis revealed that there were at least 19 candidate iron-repressible proteins in C. trachomatis and at least one protein which was iron inducible. One putative iron-repressible protein was confirmed by Western blot (immunoblot) analysis to be the chlamydial heat shock protein 60 (hsp60). The enhanced production of this antigen by chlamydiae as a result of iron limitation is of particular importance since there is a well-documented association between chlamydial hsp60 and destructive immunopathological sequelae in infected patients. PMID- 9353032 TI - Cloning, sequencing, and expression of the mig gene of Mycobacterium avium, which codes for a secreted macrophage-induced protein. AB - Mycobacterium avium is an intracellular pathogen that has evolved to be a frequent cause of disseminated infection in immunocompromised patients. Although these bacilli are readily phagocytized, they are able to survive and even multiply within human macrophages. The process whereby mycobacteria circumvent the lytic functions of the macrophages is currently not well understood, but this is a key aspect in the pathogenicity of all pathogenic mycobacteria. Previously, we identified a gene in M. avium, designated mig (for macrophage-induced gene), the expression of which is induced when the bacilli grow in human macrophages (G. Plum and J. E. Clark-Curtiss, Infect. Immun. 62:476-483, 1994). In the present study we show that (i) the nucleotide sequence of the mig gene has an open reading frame of 295 amino acids with a strong bias for mycobacterial codon usage, (ii) the mig gene also codes for a putative signal peptide of 19 amino acid residues, (iii) mig is induced by acidity to be expressed as an early secreted 30-kDa protein, and (iv) the Mig protein exhibits an AMP-binding domain signature. However, beyond this motif which is common to enzymes that activate a large variety of substrates, no homologies to known sequences are found. We also show that (v) Mycobacterium smegmatis strains expressing the Mig protein have a limited advantage for survival in macrophages. These findings may be concordant with a role of the mig gene in the virulence of M. avium. PMID- 9353033 TI - Tonsillar application of killed Streptococcus mutans induces specific antibodies in rabbit saliva and blood plasma without inducing a cross-reacting antibody to human cardiac muscle. AB - When Streptococcus mutans cells are injected into the skeletal muscle of rabbits, an antibody against human cardiac muscle, as well as an anti-S. mutans antibody, is induced in blood plasma. Our previous study showed that when sheep erythrocytes are applied to palatine tonsils, an antibody against the applied cells is induced both in blood plasma and saliva. This antibody has no activity against cardiac muscle. It is not clear, however, if S. mutans application to the tonsils evokes an antibody response against cardiac muscle. In this study, we immunized rabbits against S. mutans or Streptococcus sobrinus by tonsillar application or by intramuscular injection every 3 days for 6 weeks. Tonsillar applications of formalin-killed cells of S. mutans induced saliva immunoglobulin A (IgA) and blood plasma IgG to the applied cells. In contrast, intramuscular injection of such cells induced only blood plasma IgG. When the route of immunization was intramuscular injection, antibodies in blood plasma cross reacted with cardiac muscle. By enzyme-immunohistochemistry and Ouchterlony immunodiffusion tests, no cross-reaction to cardiac muscle was observed with the antibody in saliva or in blood plasma after the tonsillar applications. Western blotting of the S. mutans antigen showed that blood plasma from rabbits injected with S. mutans reacted with antigens of 46, 52, 62, and 85 kDa, while that from rabbits subjected to tonsillar application of S. mutans did not react with these bands. Similar results were obtained for S. sobrinus applications. Thus, tonsillar applications of mutants group streptococci induce antibodies differing in antigen specificity and do not induce any cross-reacting antibody to cardiac muscle. PMID- 9353034 TI - Direct interactions of human natural killer cells with Cryptococcus neoformans inhibit granulocyte-macrophage colony-stimulating factor and tumor necrosis factor alpha production. AB - Human natural killer (NK) cells and T lymphocytes can bind to and inhibit the growth of the yeast-like organism Cryptococcus neoformans. Binding of target cells to NK or T cells also has the potential to modulate cytokine production by the effector cells. In this study, we assessed the ability of C. neoformans to modulate NK cell production, or in some cases T-cell production, of granulocyte macrophage colony-stimulating factor (GM-CSF) or tumor necrosis factor alpha (TNF alpha). We found that freshly isolated human NK cells from most individuals make GM-CSF and TNF-alpha constitutively when cultured in vitro. The addition of C. neoformans to T-cell fractions which do not make GM-CSF constitutively did not affect GM-CSF production, but the addition of C. neoformans to NK cell fractions significantly reduced the amounts of GM-CSF produced in most NK cell samples. The reduction in the amount of GM-CSF in C. neoformans-NK cell cocultures could not be attributed to loss of lymphocyte viability or to C. neoformans adsorbing or degrading the cytokine and was dependent on direct contact between the NK cells and cryptococcal cells. GM-CSF was not the only cytokine to be down-regulated. TNF-alpha production was also diminished when NK cells were incubated with C. neoformans. The regulation of both cytokines was at the transcriptional level because GM-CSF and TNF-alpha mRNA levels were lower in NK cell samples incubated with C. neoformans than in NK cell samples incubated without C. neoformans. Diminished production of constitutively produced cytokines resulting from the interaction of NK cells with cryptococcal cells has the potential to affect phagocytic cells in the immediate regional environment and to damp the immune response. PMID- 9353035 TI - Escherichia coli strains with nonimmune immunoglobulin-binding activity. AB - We have identified several strains of Escherichia coli which contain immunoglobulin-binding activity on the cell surface. Affinity-purified antibodies ordinarily used as secondary antibodies in immunodetection protocols were bound by 6 of 72 strains of the ECOR reference collection of E. coli. The Fc fragments of both human and sheep immunoglobulin G (IgG) were also bound, demonstrating the nonimmune nature of the phenomenon. Binding of conjugated IgG Fc directly to unfixed cells was observed by fluorescence microscopy. Western blots showed that the immunoglobulin-binding material occurs in the form of multiple bands, with the apparent molecular masses of the most prominent bands exceeding 100 kDa. No two of the strains have the same pattern of bands. The binding activity in extracts was sensitive to proteinase K. The binding activity of intact cells was reduced preferentially by trypsin digestion, demonstrating exposure at the cell surface. Expression of binding activity in Luria-Bertani broth cultures was favored by a temperature of 37 degrees C and entry into stationary phase of growth. PMID- 9353036 TI - Failure to block adhesion of Plasmodium falciparum-infected erythrocytes to ICAM 1 with soluble ICAM-1. AB - The adhesion of Plasmodium falciparum-infected erythrocytes is thought to play a central role in the pathogenesis of severe malaria. ICAM-1 has been identified as one of the host receptors for parasitized erythrocytes and has been implicated as being involved in progression to cerebral malaria. Thus, intervention strategies based on the reversal of this interaction could potentially be used to reduce morbidity and mortality. We have investigated the inhibition of the interaction between ICAM-1 and infected erythrocytes by using recombinant soluble ICAM-1 as competitor and find that we are unable to reduce adhesion to ICAM-1 in vitro. PMID- 9353037 TI - Induction of an immune response by oral administration of recombinant botulinum toxin. AB - A gene encoding the full-size botulinum neurotoxin serotype C was reconstructed in vector pQE-30 and expressed at high levels in Escherichia coli. Three amino acid mutations (H229-->G, E230-->T, and H233-->N) were generated in the zinc binding motif, resulting in complete detoxification of the modified recombinant holotoxin. The PCR-amplified wild-type light chain of botulinum neurotoxin serotype C was also expressed in E. coli and used as a control in all experiments. Modified recombinant holotoxin and light chain contained a histidine affinity tag at the amino terminus, which was used for detection and purification. Recombinant proteins were purified on nickel affinity resin and analyzed by Western blotting with the anti-histidine tag and anti-neurotoxin C antibodies. The results indicated that the 150-kDa molecule of modified recombinant holotoxin and the 50-kDa recombinant light chain were synthesized without degradation; however, E. coli did not provide for efficient nicking of modified recombinant toxin. Modified recombinant holotoxin was not toxic to mice, had no effect on nerve-evoked muscle twitch in vitro, and was not able to cleave syntaxin in crude synaptosome preparations. The recombinant light chain was also nontoxic in vivo, had no effect on evoked muscle twitch, but was able to cleave syntaxin. Modified recombinant neurotoxin and light chain were administered to animals either orally or subcutaneously. Both oral administration and subcutaneous administration of modified recombinant neurotoxin evoked high levels of serum antibodies and protective immunity. Oral administration of recombinant light chain evoked no systemic response, whereas subcutaneous administration evoked antibody production and immunity. PMID- 9353038 TI - Nucleotide sequence of the Porphyromonas gingivalis W83 recA homolog and construction of a recA-deficient mutant. AB - Degenerate oligonucleotide primers were used in PCR to amplify a region of the recA homolog from Porphyromonas gingivalis W83. The resulting PCR fragment was used as a probe to identify a recombinant lambda DASH phage (L10) carrying the P. gingivalis recA homolog. The recA homolog was localized to a 2.1-kb BamHI fragment. The nucleotide sequence of this 2.1-kb fragment was determined, and a 1.02-kb open reading frame (341 amino acids) was detected. The predicted amino acid sequence was strikingly similar (90% identical residues) to the RecA protein from Bacteroides fragilis. No SOS box, characteristic of LexA-regulated promoters, was found in the 5' upstream region of the P. gingivalis recA homolog. In both methyl methanesulfonate and UV survival experiments the recA homolog from P. gingivalis complemented the recA mutation of Escherichia coli HB101. The cloned P. gingivalis recA gene was insertionally inactivated with the ermF-ermAM antibiotic resistance cassette to create a recA-deficient mutant (FLL33) by allelic exchange. The recA-deficient mutant was significantly more sensitive to UV irradiation than the wild-type strain, W83. W83 and FLL33 showed the same level of virulence in in vivo experiments using a mouse model. These results suggest that the recA gene in P. gingivalis W83 plays the expected role of repairing DNA damage caused by UV irradiation. However, inactivation of this gene did not alter the virulence of P. gingivalis in the mouse model. PMID- 9353039 TI - Toxoplasma gondii sporozoites form a transient parasitophorous vacuole that is impermeable and contains only a subset of dense-granule proteins. AB - Toxoplasma gondii sporozoites form two parasitophorous vacuoles during development within host cells, the first (PV1) during host cell invasion and the second (PV2) 18 to 24 h postinoculation. PV1 is structurally distinctive due to its large size, yet it lacks a tubulovesicular network (C. A. Speer, M. Tilley, M. Temple, J. A. Blixt, J. P. Dubey, and M. W. White, Mol. Biochem. Parasitol. 75:75-86, 1995). Confirming the finding that sporozoites have a different electron-dense-granule composition, we have now found that sporozoites within oocysts lack the mRNAs encoding the 5' nucleoside triphosphate hydrolases (NTPase). NTPase first appears 12 h postinfection. Other tachyzoite dense-granule proteins, GRA1, GRA2, GRA4, GRA5, and GRA6, were detected in oocyst extracts, and antibodies against these proteins stained granules in the sporozoite cytoplasm. In contrast to tachyzoite invasion of host cells, however, sporozoites did not exocytose the dense-granule proteins GRA1, GRA2, or GRA4 during PV1 formation. Even after NTPase induction, these proteins were retained within cytoplasmic granules rather than being secreted into PV1. Only GRA5 was secreted by the sporozoite during host cell invasion, becoming associated with the membrane surrounding PV1. Microinjection of sporozoite-infected cells with fluorescent dyes showed that PV1 is impermeable to fluorescent dyes with molecular masses as small as 330 Da, indicating that PV1 lacks channels through which molecules can pass from the host cytoplasm into the vacuole. By contrast, lucifer yellow rapidly diffused into PV2, demonstrating the presence of molecular channels. These studies indicate that PV1 and PV2 are morphologically, immunologically, and functionally distinct, and that PV2 appears to be identical to the tachyzoite vacuole. The inaccessibility of PV1 to host cell nutrients may explain why parasite replication does not occur in this vacuole. PMID- 9353040 TI - Use of a novel approach, termed island probing, identifies the Shigella flexneri she pathogenicity island which encodes a homolog of the immunoglobulin A protease like family of proteins. AB - The she gene of Shigella flexneri 2a, which also harbors the internal enterotoxin genes set1A and set1B (F. R. Noriega, GenBank accession no. U35656, 1995) encodes a homolog of the virulence-related immunoglobulin A (IgA) protease-like family of secreted proteins, Tsh, EspC, SepA, and Hap, from an avian pathogenic Escherichia coli, an enteropathogenic E. coli, S. flexneri 5, and Haemophilus influenzae, respectively. To investigate the possibility that this locus was carried on a larger deletable element, the S. flexneri 2a YSH6000T she gene was insertionally disrupted by allelic exchange using a Tn10-derived tetAR(B) cassette. Then, to detect loss of the she locus, the tetracycline-resistant derivative was plated onto fusaric acid medium to select for tetracycline-sensitive revertants, which were observed to arise at a frequency of 10(-5) to 10(-6). PCR and pulsed-field gel electrophoresis analysis confirmed loss of the she::tetAR(B) locus in six independent tetracycline-sensitive isolates. Sample sequencing over a 25-kb region flanking she identified four insertion sequence-like elements, the group II intron-like sequence Sf.IntA, and the 3' end of a second IgA protease-like homolog, sigA, lying 3.6 kb downstream and in an orientation inverted with respect to she. The deletion was mapped to chromosomal NotI fragment A and determined to have a size of 51 kb. Hybridization with flanking probes confirmed that at least 17.7 kb of the 51-kb deletable element was unique to the seven she+ strains investigated, supporting the conclusion that she lay within a large pathogenicity island. The method described in this study, termed island probing, provides a useful tool to further the study of pathogenicity islands in general. Importantly, this approach could also be of value in constructing safer live attenuated bacterial vaccines. PMID- 9353041 TI - Detoxification of the Helicobacter pylori cytotoxin. AB - Treatment of the Helicobacter pylori vacuolating cytotoxin with very low concentrations of formaldehyde resulted in abrogation of toxic activity in both a HeLa cell vacuolation assay and an in vivo assay of gastric epithelial damage. Detoxification had only a minimal effect on the integrity of the oligomeric or monomeric structure. The toxoid retained the ability to bind to target cells and to induce high-titer neutralizing antibodies after immunization of rabbits. Furthermore, oral immunization of mice with the toxoid resulted in protection against infective challenge with mouse-adapted strains of H. pylori. The sensitivity of the toxin to formaldehyde treatment suggests that a few lysine residues in the protein may be essential for toxic activity and that VacA detoxified in this manner may be a potential candidate for inclusion in a vaccine against H. pylori infection and disease. PMID- 9353042 TI - Chemokine secretion by human polymorphonuclear granulocytes after stimulation with Mycobacterium tuberculosis and lipoarabinomannan. AB - Macrophages (MAC) and polymorphonuclear granulocytes (PNG) are professional phagocytes which perform essential functions in antibacterial defense. The intracellular bacterium Mycobacterium tuberculosis persists and replicates in resting macrophages. Although it is generally assumed that activated MAC are central to protection against M. tuberculosis, PNG may also contribute to defense. We wondered whether PNG produce proinflammatory chemokines after stimulation by M. tuberculosis or its major cell wall component, lipoarabinomannan (LAM). In this study, we showed that M. tuberculosis- and LAM activated human PNG secrete the leukocyte attractant interleukin-8 (IL-8) and the PNG-specific chemokine GRO-alpha in a dose-dependent manner. Treatment of PNG with the leukotriene-B4 inhibitor MK-886 prior to stimulation with M. tuberculosis or LAM partially blocked IL-8 and GRO-alpha induction, suggesting involvement of the 5-lipoxygenase pathway in the secretion of these chemokines. We conclude that PNG contribute to early resistance to M. tuberculosis via chemokine secretion. PMID- 9353043 TI - Salmonellae activate tumor necrosis factor alpha production in a human promonocytic cell line via a released polypeptide. AB - Invasive strains of Salmonella spp. cause both systemic and localized infections in humans. The ability to resist infection and some aspects of the tissue pathology associated with the presence of Salmonella in the gastrointestinal tract have been shown to be mediated in part by the induction of tumor necrosis factor alpha (TNF-alpha), a proinflammatory cytokine produced by activated macrophages and lymphocytes. Recent reports indicate that TNF-alpha is involved in the induction of human immunodeficiency virus replication by Salmonella in the latently infected human promonocytic cell line U1. In the present study, we investigated the effects of Salmonella on TNF-alpha production in U1 cells and a related cell line, U38. Unlike Escherichia coli or Yersinia enterocolitica, salmonellae rapidly induce TNF-alpha expression in these cells through a released factor(s). Time course experiments show that the kinetics of TNF-alpha production by U38 cells stimulated with Salmonella conditioned medium closely resemble those observed in response to live Salmonella. The observation that TNF-alpha levels are elevated by 60 min after exposure to either bacteria or their conditioned medium suggests that the soluble inducer is continuously released or shed by the bacteria and that the signal acts rapidly to increase TNF-alpha production. Furthermore, the ability to produce the TNF-alpha inducer is shared by at least four Salmonella serotypes and does not correlate with the abilities to invade and to survive within phagocytes. Treatment of active conditioned medium with trypsin, but not low pH, high temperature, or urea, significantly inhibits its TNF-alpha-inducing effect on U38 cells, a finding which points to a polypeptide product of Salmonella as the mediator of TNF-alpha production. Gel filtration chromatography of Salmonella conditioned medium reveals two peaks of activity, consistent with molecular masses of approximately 150 and 110 kDa. PMID- 9353044 TI - Transcription of the Corynebacterium diphtheriae hmuO gene is regulated by iron and heme. AB - The hmuO gene is required for the utilization of heme and hemoglobin as iron sources by Corynebacterium diphtheriae. The product of hmuO has homology to eukaryotic heme oxygenases which are involved in the degradation of heme and the release of iron. To investigate the mechanism of hmuO regulation, a promoterless lacZ gene present on the promoter-probe vector pCM502 was placed under transcriptional control of the hmuO promoter. In C. diphtheriae C7, optimal expression from the hmuO promoter was obtained only in the presence of heme or hemoglobin under low-iron conditions. Expression of hmuO in high-iron medium containing heme was repressed five- to sixfold from that seen under low-iron conditions in the presence of heme. Transcription from the hmuO promoter in the absence of heme or hemoglobin was fully repressed in high-iron medium and was expressed at very low levels in iron-depleted conditions. Expression studies with tile hmuO-lacZ fusion construct in C7hm723, a dtxR mutant of C7, and in a hmuO mutant of C. diphtheriae HC1 provided further evidence that transcription of the hmuO promoter is repressed by DtxR and iron and activated by heme. In Escherichia coli, the hmuO promoter was expressed at very low levels under all conditions examined. Gel mobility shift assays and DNase I footprinting experiments indicated that DtxR binds in a metal-dependent manner to a sequence that overlaps the putative hmuO promoter. Total cellular RNA isolated from C. diphtheriae was used to identify the transcriptional start site for the hmuO gene. Northern blot analysis suggested that the hmuO mRNA was monocistronic and that transcription was heme inducible. PMID- 9353045 TI - Attenuation, persistence, and vaccine potential of an Edwardsiella ictaluri purA mutant. AB - In this study, an adenine-auxotrophic strain of Edwardsiella ictaluri was constructed and its virulence, tissue persistence, and vaccine efficacy were evaluated. A clone containing the purA gene was isolated from an E. ictaluri genomic library, sequenced, and shown to have an overall sequence identity of 79.3% at the nucleotide level and 85.7% at the amino acid level with the Escherichia coli purA gene. The cloned E. ictaluri purA gene was mutated by deleting a 598-bp segment of the gene and inserting the kanamycin resistance gene from Tn903 into the gap. The delta purA::Km(r) gene was subcloned into the suicide plasmid pGP704, and the resulting plasmid was used to deliver the modified gene into a virulent strain of E. ictaluri by conjugation. Homologous recombination replaced the chromosomal purA gene with the mutated gene to create an adenine-auxotrophic strain (LSU-E2). Compared to wild-type E. ictaluri, LSU-E2 was highly attenuated by the injection, immersion, and oral routes of exposure. By the injection route, LSU-E2 had a 50% lethal dose (LD50) that was greater than 5 logs10 higher than the LD50 for wild-type E. ictaluri. In a tissue persistence study, LSU-E2 was able to invade channel catfish by the immersion route and persist in internal organs for at least 48 h. Channel catfish that were vaccinated with a single immersion dose of LSU-E2 had mortality significantly lower (P < 0.01) following a wild-type E. ictaluri challenge than that of nonvaccinated fish. PMID- 9353046 TI - Hyperproduction of alpha-toxin by Staphylococcus aureus results in paradoxically reduced virulence in experimental endocarditis: a host defense role for platelet microbicidal proteins. AB - Staphylococcal alpha-toxin targets several cell types which are important components of cardiac vegetations in endocarditis, including platelets, erythrocytes, and endothelial cells. We evaluated the in vivo role of Staphylococcus aureus alpha-toxin in experimental endocarditis by using isogenic strains differing in the capacity to produce functional alpha-toxin, including 8325-4 (wild-type strain), DU-1090 (a mutant strain with allelic replacement of the alpha-toxin gene [hla]), DU1090(pH35L) (a mutant strain producing a target cell-binding but nonlytic toxin), DU1090(pDU1212) (a variant of DU1090 carrying the cloned hla gene on a multicopy plasmid), and DU1090(pCL84::hla) (a variant of DU1090 with a single copy of the hla gene cloned into the chromosomal lipase locus). In vitro, wild-type alpha-toxin (from parental strain 8325-4) extensively lysed both erythrocytes and platelets. In contrast, mutant alpha-toxin [from strain DU1090(pH35L)] lysed neither cell type. Following exposure to the wild type alpha-toxin, platelet lysates were found to contain microbicidal activity against Bacillus subtilis (but not against Micrococcus luteus), as well as against the parental and alpha-toxin variant S. aureus strains noted above. Furthermore, lysate microbicidal activity was heat stable, neutralized by polyanionic filters or compounds, and recoverable from anionic filter membranes by hypertonic saline elution. These characteristics are consistent with those of cationic platelet microbicidal proteins (PMPs). Reverse-phase high-pressure liquid chromatography and polyacrylamide gel electrophoresis confirmed the presence of three distinct PMPs (1, 2, and 3) in platelet lysates. In experimental endocarditis, the two variant staphylococcal strains producing either minimal alpha-toxin or nonlytic alpha-toxin in vitro [strains DU1090 and DU1090(pH35L), respectively] exhibited significantly lower virulence in vivo than the parental strain (decreased intravegetation staphylococcal densities). Paradoxically, the two variant staphylococcal strains producing alpha-toxin at supraparental levels in vitro [strains DU1090(p1212) and DU1090(pCL84::hla)] also exhibited significantly decreased induction rates and intravegetation staphylococcal densities in experimental endocarditis versus the parental strain. The reduced in vivo virulence of the latter variant staphylococcal strains could not be explained by differences in bacteremic clearance or initial adherence to sterile vegetations (compared to the parental strain). These findings suggest that the reduced virulence exhibited by the variant staphylococcal strains in this model was related to pathogenetic events subsequent to bacterial adherence to the damaged endocardium. Excess intravegetation secretion of alpha-toxin, leading to increased PMP release (secondary to either increased platelet secretion or lysis), may well explain the reduced virulence observed in experimental endocarditis. PMID- 9353047 TI - Borrelia burgdorferi strain-specific Osp C-mediated immunity in mice. AB - Antibodies to the outer surface proteins (Osps) A, B, and C of the spirochete Borrelia burgdorferi can prevent infection in animal models of Lyme borreliosis. We have previously demonstrated that immune serum from mice infected with B. burgdorferi N40 can also prevent challenge infection and induce disease regression in infected mice. The antigens targeted by protective and disease modulating antibodies are presently unknown, but they do not include Osp A or Osp B. Because Osp C antibodies are present in immune mouse serum, we investigated the ability of hyperimmune serum to recombinant Osp C (N40) to protect mice against challenge infection with N40 spirochetes. In both active and passive immunization studies, Osp C (N40) antiserum failed to protect mice from challenge infection with cultured organisms. Mice actively immunized with recombinant Osp C (N40) were susceptible to tick-borne challenge infection, and nymphal ticks remained infected after feeding on Osp C-hyperimmunized mice. In contrast, similar immunization studies performed with Osp C (PKo) antiserum prevented challenge infection of mice with a clone of PKo spirochetes pathogenic for mice. Both Osp C (N40) and Osp C (PKo) antisera showed minimal in vitro borreliacidal activity, and immunofluorescence studies localized Osp C beneath the outer membrane of both N40 and PKo spirochetes. We conclude that Osp C antibody mediated immunity is strain specific and propose that differences in Osp C surface expression by spirochetes in vivo may account for strain-specific immunity. PMID- 9353048 TI - Catalase, a novel antigen for Helicobacter pylori vaccination. AB - The efficacy of an orogastric vaccine comprised of purified Helicobacter pylori catalase plus the mucosal adjuvant cholera toxin (CT) was examined with both the Helicobacter felis and H. pylori mouse models with BALB/c mice. Native H. pylori catalase (200 microg) plus CT was initially used as a vaccine antigen in the H. felis mouse model and protected 80% (8 of 10) of the challenged animals, while all control animals were infected (20 of 20). In a follow-up experiment, recombinant H. pylori catalase plus CT was used for immunization, and groups of mice were challenged with the Sydney strain of H. pylori. Immunization with recombinant catalase protected a significant proportion (9 of 10) of the mice from H. pylori challenge, indicating that this enzyme should be considered as a candidate for a future vaccine. This study provides the first available data on the efficacy of protective immunization with the new Sydney strain of H. pylori in a mouse model. These data also provide indirect evidence that proteins which are normally intracellular, such as catalase, may be present on the surface of H. pylori and thus may provide targets for immunization. PMID- 9353049 TI - Phase variation and conservation of lipooligosaccharide epitopes in Haemophilus somnus. AB - The bovine-specific pathogen Haemophilus somnus is capable of undergoing structural and antigenic phase variation in its lipooligosaccharide (LOS) components after in vivo and in vitro passage. However, commensal isolates from the reproductive tract have not been observed to vary in phase (T. J. Inzana, R. P. Gogolewski, and L. B. Corbeil, Infect. Immun. 60:2943-2951, 1992). We now report that specific monoclonal antibodies (MAbs) to the LOSs of Haemophilus aegyptius, Neisseria gonorrhoeae, and Haemophilus influenzae, as well as H. somnus, reacted with some phase-variable epitopes in H. somnus LOS. All reactive MAbs bound to LOS components of about 4.3 kDa in the same H. somnus isolates, including a non-phase-varying strain. Following in vitro passage of a clonal variant of strain 738 that was nonreactive with the MAbs, 11.8% of young colonies shifted to a reactive phenotype. A digoxigenin-labelled 5' CAATCAATCAATCAATCAATCAATCAAT-3' oligonucleotide probe hybridized to genomic DNA from strain 738 but did not react with DNA from a non-phase-varying strain. Sequence analysis of the gene containing 5'-CAAT-3' tandem sequences revealed 48% amino acid homology with the lex-2B gene-encoded protein of H. influenzae type b. Our results indicate that some LOS epitopes are conserved between H. somnus and other Haemophilus and Neisseria species, that LOS phase variation may occur at a high rate in some strains of H. somnus, and that phase variation may, in part, be due to 5'-CAAT-3' tandem sequences present in H. somnus genes. PMID- 9353050 TI - Gamma interferon induces Fas-dependent apoptosis of Peyer's patch T cells in mice following peroral infection with Toxoplasma gondii. AB - Since we previously observed a remarkable decrease in the numbers of T cells in the Peyer's patches of the small intestines in C57BL/6 mice following peroral infection with Toxoplasma gondii, we performed studies to examine the mechanism(s) whereby this decrease in numbers of the T cells occurs. We found that apoptotic cell death of CD4+ and CD8+ alphabeta T cells occurred in Peyer's patches following infection. Upregulation of Fas expression was observed in these T cells. C57BL/6-background mutant mice which lack functional Fas antigen did not develop apoptosis in their Peyer's patches following infection. Treatment of infected C57BL/6 mice with anti-gamma interferon (IFN-gamma) monoclonal antibodies prevented the upregulation of Fas on their Peyer's patch T cells and inhibited the occurrence of apoptosis of these T cells. These results indicate that IFN-gamma induces Fas-dependent apoptosis in CD4+ and CD8+ alphabeta T cells in Peyer's patches in C57BL/6 mice following peroral infection with T. gondii. PMID- 9353051 TI - Opsonization of Actinobacillus actinomycetemcomitans by immunoglobulin G antibodies to the O polysaccharide of lipopolysaccharide. AB - Sera of localized juvenile periodontitis (LJP) patients colonized by Actinobacillus actinomycetemcomitans serotype b often contain markedly elevated levels of immunoglobulin G (IgG) antibodies to serospecific determinants in the O polysaccharide of lipopolysaccharide (LPS), as well as to outer membrane proteins of this species. IgG antibodies in LJP sera are known to opsonize A. actinomycetemcomitans for subsequent phagocytosis and killing by human neutrophils. The objective of this study was to determine whether outer membrane proteins or serospecific determinants in LPS are the primary target for opsonic IgG antibodies in LJP sera. An A. actinomycetemcomitans serotype b O polysaccharide affinity column was constructed and subsequently used to purify LPS-specific IgG antibodies from LJP serum. The affinity-purified anti-LPS IgG antibodies were enriched in content of IgG2 (66.2%, compared with 37.0% in the total IgG fraction) and were immunospecific for A. actinomycetemcomitans serotype b LPS. In an opsonophagocytic assay using neutrophils from donors who were homozygous for the H131 allotype of Fcy receptor IIa (CD32), it was found that LPS-specific IgG antibodies exhibited substantially greater opsonic activity toward A. actinomycetemcomitans serotype b than an LJP IgG fraction that was depleted of LPS-reactive antibodies but contained antibodies against outer membrane proteins of this species. The results of this study indicate that serospecific determinants in the O polysaccharide of A. actinomycetemcomitans serotype b are a principal target for opsonic antibodies in sera of LJP subjects. PMID- 9353052 TI - Transcription of genes encoding iron and heme acquisition proteins of Haemophilus influenzae during acute otitis media. AB - Unencapsulated Haemophilus influenzae is the second most common etiologic agent of otitis media in children. H. influenzae requires heme for aerobic growth in vitro and is able to utilize hemoglobin and complexes of heme-hemopexin, heme albumin, and hemoglobin-haptoglobin and ferritransferrin as sources of iron and heme in vitro. Several of the acquisition mechanisms have been characterized and been shown to be heme repressible in vitro. However, little is known about the expression of heme and/or iron acquisition mechanisms during infections in the middle ear. This study was performed to determine if the genes encoding heme and iron acquisition proteins are transcribed during in vivo growth and to compare these findings with those for samples grown in vitro. Reverse transcriptase PCR (RT-PCR) was used to analyze total RNA fractions derived from in vitro- and in vivo-grown H. influenzae. Genes encoding the transferrin-binding proteins TbpA and TbpB, the 100-kDa hemopexin-binding protein HxuA, and the hemoglobin-binding protein HgpA were transcribed during otitis media. Twelve middle ear fluid samples were analyzed by blind RT-PCR to determine the transcriptional status of these genes in H. influenzae during otitis media. Five isolates had transcripts corresponding to tbpA, tbpB, and hxuA. The presence of hgpA transcripts was variable, depending on the presence of hgpA in the genome of the H. influenzae isolate. Samples without H. influenzae gene transcripts contained other etiologic agents commonly causing otitis media. These data demonstrate that H. influenzae iron and/or heme acquisition genes are transcribed during otitis media and suggest that the microenvironment during acute otitis media starves H. influenzae of heme. PMID- 9353053 TI - Development of a model of low-inoculum Streptococcus pneumoniae intrapulmonary infection in infant rats. AB - We have developed a model of low-inoculum Streptococcus pneumoniae infection in infant rats. We challenged 4-day-old Sprague-Dawley pups via intraperitoneal or intrapulmonary injection of S. pneumoniae serotypes 1, 3, 4, 5, 6b, 7f, 9v, 14, 19f, and 23f. To achieve bacteremia with low inocula, it was necessary to passage the isolates in rats. Inocula of the 10 S. pneumoniae serotypes producing bacteremia in 50% or more animals ranged from 1 to 400 CFU. Virulence was similar by intraperitoneal and intrapulmonary routes. Lung specimens from animals challenged by the intrapulmonary route grew S. pneumoniae and demonstrated histologic evidence of focal infection. Meningitis was detected in 20 to 50% of bacteremic animals, and mortality invariably followed bacteremia within 24 to 48 h. This model of intrapulmonary infection uses low inocula of S. pneumoniae and results in bacteremia, meningitis, and death in infant rats. PMID- 9353054 TI - Contribution of the Mn-cofactored superoxide dismutase (SodA) to the virulence of Yersinia enterocolitica serotype O8. AB - Enteric pathogens harbor a set of enzymes (e.g., superoxide dismutases [SOD]) for detoxification of endogenous and exogenous reactive oxygen species which are encountered during infection. To analyze the role of the Mn-cofactored SOD (SodA) in the pathogenicity of yersiniae, we cloned the sodA gene of Yersinia enterocolitica serotype O8 by complementation of an Escherichia coli sodA sodB mutant and subsequently constructed an isogenic mutant by allelic exchange. Sequence analysis revealed an open reading frame that enabled the deduction of a sequence of 207 amino acids with 85% identity to SodA of E. coli. In a mouse infection model, the sodA null mutant was strongly attenuated in comparison to its parental strain. After intravenous infection, the survival and multiplication of the mutant in the spleen and liver were markedly reduced. In contrast, inactivation of sodA had only minor effects on survival and multiplication in the gut and Peyer's patches, as could be demonstrated in the orogastric infection model. The reduction in virulence was accompanied by a low but significant increase of susceptibility of the soda mutant to bacterial killing by polymorphonuclear leukocytes (PMN) and an alteration of the intracellular chemiluminescence response of PMN. These results suggest that the resistance of Y. enterocolitica to exogenous oxygen radicals produced by phagocytes involves the Mn-cofactored SOD. The important role of sodA for the pathogenicity of Y. enterocolitica could also be due to detoxification of endogenous, metabolically produced oxygen radicals which are encountered by extracellular enteric pathogens during the invasion of the host. PMID- 9353055 TI - Differences in coughing and other responses to intrabronchial infection with Bordetella pertussis among strains of rats. AB - Four strains of rats were each infected intrabronchially with approximately 10(8) CFU of Bordetella pertussis 18-323 encased in fine agarose beads. After 8 days, Sprague-Dawley rats developed the highest incidence of coughing paroxysms, as monitored with voice-activated tape recorders; Brown Norway, Lewis, and Hooded Lister rats coughed significantly less frequently. Marked leukocytosis, with counts up to four times the normal levels, and retardation of normal weight gain occurred in all four rat strains. Both coughing and leukocytosis were greater in animals that were infected at 4 weeks of age than in those infected at 6 weeks of age. Total serum immunoglobulin E (IgE) rose in all four rat strains 9- to 244 fold by day 8 after infection and returned to near preinfection levels at 6 weeks. Sprague-Dawley and Lewis rats, which had the lowest basal levels of total IgE in serum, showed the greatest degrees of elevation. All four rat strains had IgG to B. pertussis whole-cell sonicate and to filamentous hemagglutinin in 6 week-postinfection sera. However, the strains differed in production of IgG to pertussis toxin, with Sprague-Dawley rats having the highest titers and Hooded Lister and Lewis rats being nonresponders. These studies highlight the importance of rat strain as a variable in the coughing-rat model of pertussis and validate the choice of the Sprague-Dawley rats in previous studies. PMID- 9353056 TI - Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus. AB - Aspergillus fumigatus possesses two catalases (described as fast and slow on the basis of their electrophoretic mobility). The slow catalase has been recognized as a diagnostic antigen for aspergillosis in immunocompetent patients. The antigenic catalase has been purified. The enzyme is a tetrameric protein composed of 90-kDa subunits. The corresponding cat1 gene was cloned, and sequencing data show that the cat1 gene codes for a 728-amino-acid polypeptide. A recombinant protein expressed in Pichia pastoris is enzymatically active and has biochemical and antigenic properties that are similar to those of the wild-type catalase. Molecular experiments reveal that CAT1 contains a signal peptide and a propeptide of 15 and 12 amino acid residues, respectively. cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain. In addition, there was no difference in pathogenicity between the cat1 mutant and its parental cat1+ strain in a murine model of aspergillosis. PMID- 9353057 TI - Exclusion of bioactive contaminations in Streptococcus pyogenes erythrogenic toxin A preparations by recombinant expression in Escherichia coli. AB - The streptococcal erythrogenic exotoxin A (SPEA) belongs to the family of bacterial superantigens and has been implicated in the pathogenesis of a toxic shock-like syndrome and scarlet fever. Concerning its biological activity, mainly T-cell-stimulatory properties, conflicting data exist. In this study, we show that most of the SPEA preparations used so far contain biologically active contaminations. Natural SPEA from the culture supernatant of Streptococcus pyogenes NY-5 and recombinant SPEA purified from the culture filtrate of S. sanguis are strongly contaminated with DNases. We show that natural SPEA induces more tumor necrosis factor alpha (TNF-alpha) than recombinant SPEA, but we also show that DNases are able to induce TNF-alpha. In commercial SPEA preparations, we identified a highly active protease, which was shown not to be SPEB. To exclude these contaminations, we overexpressed SPEA cloned in the effective high level expression vector pIN-III-ompA2 in Escherichia coli. The expressed SPEA shows the same amino acid composition as natural SPEA, whereas functional studies reported so far were carried out with toxins containing an incorrect amino terminus. We describe the rapid purification of lipopolysaccharide-, DNase-, and protease-free SPEA in two steps from the host's periplasm and its structural characterization by circular dichroism. Our results represent for the first time the production in E. coli of recombinant SPEA with the authentic N-terminal sequence and a proven superantigenic activity. Collectively, our results indicate that immunological studies of superantigens require highly purified substances free of biologically active contaminations. PMID- 9353058 TI - Treatment with homodimeric interleukin-12 (IL-12) p40 protects mice from IL-12 dependent shock but not from tumor necrosis factor alpha-dependent shock. AB - The role of interleukin-12 (IL-12) was investigated in different shock models using anti-IL-12 reagents. IL-12 is composed of two disulfide-bonded subunits, p35 and p40. The IL-12 p40 homodimer (p40)2 has been shown to be a potent IL-12 antagonist in vitro. We investigated its in vivo inhibitory capacity in different shock models of mice. We could demonstrate that (p40)2 is able to protect mice from septic shock in primarily IL-12-dependent models such as the Shwartzman reaction and lipopolysaccharide (LPS)-induced shock, whereas (p40)2 has no effect in the tumor necrosis factor alpha-dependent LPS/D-GalN shock model. In IL-12 dependent shock models, (p40)2 inhibits IL-12-induced gamma interferon production and thereby interferes with the cascade of cytokine release, finally leading to death. PMID- 9353059 TI - Utilization of similar mechanisms by Legionella pneumophila to parasitize two evolutionarily distant host cells, mammalian macrophages and protozoa. AB - The Legionnaires' disease bacterium, Legionella pneumophila, is an intracellular pathogen of humans that is amplified in the environment by intracellular multiplication within protozoa. Within both evolutionarily distant hosts, the bacterium multiplies in a rough endoplasmic reticulum-surrounded phagosome that is retarded from maturation through the endosomal-lysosomal degradation pathway. To gain an understanding of the mechanisms utilized by L. pneumophila to invade and replicate within two evolutionarily distant hosts, we isolated a collection of 89 mini-Tn10::kan insertion mutants that exhibited defects in cytotoxicity, intracellular survival, and replication within both U937 macrophage-like cells and Acanthamoeba polyphaga. Interestingly, the patterns of defects in intracellular survival and replication of the mutants within both host cells were highly similar, and thus we designated the defective loci in these mutants pmi (for protozoan and macrophage infectivity loci). On the basis of their ability to attach to host cells and their growth kinetics during the intracellular infection, the mutants were grouped into five groups. Groups 1 and 2 included 41 mutants that were severely defective in intracellular survival and were completely or substantially killed during the first 4 h of infection in both host cells. Three members of group 1 were severely defective in attachment to both U937 cells and A. polyphaga, and another four mutants of group 1 exhibited severe defects in attachment to A. polyphaga but only a mild reduction in their attachment to U937 cells. Four members of groups 1 and 2 were serum sensitive. Intracellular replication of mutants of the other three groups was less defective than that of mutants of groups 1 and 2, and their growth kinetics within both host cells were similar. The mutants were tested for several other phenotypes in vitro, revealing that 14 of the pmi mutants were resistant to NaCl, 3 had insertions in dot or icm, 3 were aflagellar, 12 were highly intolerant to a hyperosmotic medium, and one failed to grow in a minimal medium. Our data indicated that similar mechanisms are utilized by L. pneumophila to replicate within two evolutionarily distant hosts. Although some mechanisms of attachment to both host cells were similar, other distinct mechanisms were utilized by L. pneumophila to attach to A. polyphaga. Our data supported the hypothesis that preadaptation of L. pneumophila to infection of protozoa may play a major role in its ability to replicate within mammalian cells and cause Legionnaires' disease. PMID- 9353060 TI - Azurophilic granules of human neutrophils contain CD14. AB - CD14, the leukocyte receptor for lipopolysaccharide (LPS), is important in the response of human polymorphonuclear leukocytes (PMNs) to infection with gram negative bacteria. The level of CD14 on the PMN surface increases after exposure to some inflammatory stimuli such as N-formyl-methionyl-leucyl-phenylalanine (fMLP). These newly expressed CD14 molecules probably come from an intracellular pool of preformed receptors. We sought to further characterize PMN CD14 expression, upregulation, and shedding and to define the intracellular location of CD14 molecules. Our results demonstrate that both LPS and fMLP significantly increased CD14 cell surface expression; however, neither phorbol myristate acetate (PMA) or A23187 increased receptor levels on the PMN surface. Neither fMLP, PMA, or A23187 stimulated the release of soluble CD14 from PMNs. Intracellular CD14 was observed in >90% of PMNs examined by flow cytometry and confocal microscopy. Additional analyses using CD14 enzyme-linked immunosorbent assays and electron microscopy studies, examining PMN granules separated by discontinuous sucrose or Percoll gradients, showed that CD14 was present in both the plasma membrane-secretory vesicle fractions and azurophilic granules. PMID- 9353061 TI - Uptake of pathogenic intracellular bacteria into human and murine macrophages downregulates the eukaryotic 26S protease complex ATPase gene. AB - A differential PCR technique detected the transcriptional downregulation of the mss1 (mammalian suppressor of svg1) gene in murine J774A.1 macrophages following uptake of Salmonella typhimurium. This downregulation was also noted after entry of virulent strains of Listeria monocytogenes and Shigella flexneri, two other facultative intracellular bacterial species. In contrast, uptake of nonpathogenic Escherichia coli HB101, an aroA mutant of S. typhimurium, an invasion plasmid antigen B (ipaB) mutant of S. flexneri, hemolysin (hly) and positive-regulatory factor (prfA) mutants of L. monocytogenes, or latex beads produced mss1 expression levels similar to that of uninfected macrophages. Transcriptional downregulation of mss1 was also shown to occur in S. typhimurium-infected human U937 cells, albeit to an extent less than that in murine J774A.1 cells. In addition to a lower abundance of mss1 transcripts, we also demonstrate for the first time that less MSS1 protein was detected in intracellular-bacterium infected cells (beginning about 1 h after entry of the pathogenic intracellular bacteria) than in noninfected cells. Some strains with specific mutations in characterized genes, such as an ipaB mutant strain of S. flexneri and an hly mutant strain of L. monocytogenes, did not elicit this lower level of expression of MSS1 protein. The decrease in MSS1 within infected macrophages resulted in an accumulation of ubiquitinated proteins, substrates for MSS1. Since MSS1 comprises the ATPase part of the 26S protease that degrades ubiquitinated proteins, we hypothesize that downregulation of the mss1 gene by intracellular bacterial entry may help subvert the host cell's normal defensive response to internalized bacteria, allowing the intracellular bacteria to survive. PMID- 9353062 TI - Profiles of healing and nonhealing Cryptosporidium parvum infection in C57BL/6 mice with functional B and T lymphocytes: the extent of gamma interferon modulation determines the outcome of infection. AB - This study describes healing and nonhealing models of Cryptosporidium parvum infection with adult mice that have functional T and B lymphocytes. In our nonhealing model, mice on a C57BL/6 background which have a targeted disruption in the gamma interferon (IFN-gamma) gene (GKO mice) are utilized. C. parvum infected GKO mice shed extremely high levels of oocysts and displayed overwhelming infection of the entire small intestine. The majority of these mice succumbed within 2 to 3 weeks due to severe acute infection and profound mucosal destruction. In our healing murine model, C57BL/6J mice treated with a single injection of the neutralizing anti-IFN-gamma monoclonal antibody XMG 1.2 prior to infection were used. These mice developed two peaks of oocyst shedding but were ultimately free of parasites on day 30 of infection. Again, the small intestine was the primary site of infection. Mesenteric lymph node (MLN) cells isolated from C. parvum-infected nonhealing GKO mice proliferated and secreted interleukin 2 (IL-2) but not IFN-gamma or IL-4 in response to ex vivo restimulation with intact C. parvum sporozoites or a C. parvum sporozoite antigen preparation. In contrast, parasite-specific MLN cells isolated from healing C57BL/6J mice secreted IL-2 and IFN-gamma but not IL-4. These results suggest that IFN-gamma, either directly or indirectly, is important for resistance to and resolution of cryptosporidiosis. Moreover, these models now allow the analysis of parasite specific cell-mediated and humoral mucosal immune responses to determine what constitutes protective immunity to C. parvum. PMID- 9353063 TI - Display of a PorA peptide from Neisseria meningitidis on the bacteriophage T4 capsid surface. AB - The exterior of bacteriophage T4 capsid is coated with two outer capsid proteins, Hoc (highly antigenic outer capsid protein; molecular mass, 40 kDa) and Soc (small outer capsid protein; molecular mass, 9 kDa), at symmetrical positions on the icosahedron (160 copies of Hoc and 960 copies of Soc per capsid particle). Both these proteins are nonessential for phage infectivity and viability and assemble onto the capsid surface after completion of capsid assembly. We developed a phage display system which allowed in-frame fusions of foreign DNA at a unique cloning site in the 5' end of hoc or soc. A DNA fragment corresponding to the 36-amino-acid PorA peptide from Neisseria meningitidis was cloned into the display vectors to generate fusions at the N terminus of Hoc or Soc. The PorA-Hoc and PorA-Soc fusion proteins retained the ability to bind to the capsid surface, and the bound peptide was displayed in an accessible form as shown by its reactivity with specific monoclonal antibodies in an enzyme-linked immunosorbent assay. By employing T4 genetic strategies, we show that more than one subtype specific PorA peptide can be displayed on the capsid surface and that the peptide can also be displayed on a DNA-free empty capsid. Both the PorA-Hoc and PorA-Soc recombinant phages are highly immunogenic in mice and elicit strong antipeptide antibody titers even with a weak adjuvant such as Alhydrogel or no adjuvant at all. The data suggest that the phage T4 hoc-soc system is an attractive system for display of peptides on an icosahedral capsid surface and may emerge as a powerful system for construction of the next generation multicomponent vaccines. PMID- 9353064 TI - Study of the role of the htrB gene in Salmonella typhimurium virulence. AB - We have undertaken a study to investigate the contribution of the htrB gene to the virulence of pathogenic Salmonella typhimurium. An htrB::mini-Tn10 mutation from Escherichia coli was transferred by transduction to the mouse-virulent strain S. typhimurium SL1344 to create an htrB mutant. The S. typhimurium htrB mutant was inoculated into mice and found to be severely limited in its ability to colonize organs of the lymphatic system and to cause systemic disease in mice. A variety of experiments were performed to determine the possible reasons for this loss of virulence. Serum killing assays revealed that the S. typhimurium htrB mutant was as resistant to killing by complement as the wild-type strain. However, macrophage survival assays revealed that the S. typhimurium htrB mutant was more sensitive to the intracellular environment of murine macrophages than the wild-type strain. In addition, the bioactivity of the lipopolysaccharide (LPS) of the htrB mutant was reduced compared to that of the LPS from the parent strain as measured by both a Limulus amoebocyte lysate endotoxin quantitation assay and a tumor necrosis factor alpha bioassay. These results indicate that the htrB gene plays a role in the virulence of S. typhimurium. PMID- 9353065 TI - Integrin CR3 mediates the binding of nonspecifically opsonized Borrelia burgdorferi to human phagocytes and mammalian cells. AB - Like other pathogens, the spirochete Borrelia burgdorferi, the agent of Lyme disease, possesses multiple pathways for cell binding; adhesion to phagocytic cells is of particular interest since it reportedly occurs even in the absence of specific antibodies. This study sets out to investigate how B. burgdorferi binds to human polymorphonuclear leukocytes (PMNs) when an exogenous complement is added and how the CR3 complement receptor, known as Mac-1 or alpha(m)beta2 integrin, is involved in the binding process. Experiments performed on PMNs and CHO Mac-1-expressing cells demonstrate that binding is inhibited by monoclonal anti-iC3b site antibodies, fibrinogen, and N-acetyl-D-glucosamine. These findings, which are not present with non-Mac-transfected CHO cells, indicate that the integrin alpha(m)beta2 acts as a receptor for spirochetes in nonimmune phagocytosis; furthermore, binding occurs on different domains of the CD11b subunit, involving the iC3b site and the lectin domain. The interaction of B. burgdorferi with alpha(m)beta2 integrin adds a novel pathway to Borrelia phagocyte binding; not only does this binding affect the early stages of phagocytosis, but also it can influence the effector intracellular mechanisms which are activated by the beta2 integrin, as are the cytotoxic mechanisms. PMID- 9353066 TI - The major surface glycoprotein of Pneumocystis carinii induces release and gene expression of interleukin-8 and tumor necrosis factor alpha in monocytes. AB - Recent studies suggest that interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) may play a central role in host defense and pathogenesis during Pneumocystis carinii pneumonia. In order to investigate whether the major surface antigen (MSG) of human P. carinii is capable of eliciting the release of IL-8 and TNF-alpha, human monocytes were cultured in the presence of purified MSG. MSG stimulated cells released significant amounts of IL-8 within 4 h, and at 20 h, cells stimulated with MSG released 45.5 +/- 9.3 ng of IL-8/ml versus 3.7 +/- 1.1 ng/ml for control cultures (P = 0.01). In a similar fashion, MSG elicited release of TNF-alpha. Initial increases were also seen at 4 h, and at 20 h, TNF-alpha levels reached 6.4 +/- 1.1 ng/ml, compared to 0.08 +/- 0.01 ng/ml for control cultures (P < 0.01). A concentration-dependent increase in IL-8 and TNF-alpha secretion was observed at 20 h with 0.2 to 5 microg of MSG/ml (P < 0.01). Secretion of IL-8 and TNF-alpha from MSG-stimulated monocytes at 20 h was inhibited by 60 and 86%, respectively, after coincubation with soluble yeast mannan (P = 0.01). With an RNase protection assay, increases in steady-state mRNA levels for IL-8 and TNF-alpha were detectable at 4 h. These data show that recognition of MSG by monocytes involves a mannose-mediated mechanism and results in the release of the proinflammatory cytokines IL-8 and TNF-alpha. PMID- 9353068 TI - Serum antibodies to Porphyromonas gingivalis block the prostaglandin E2 response to lipopolysaccharide by mononuclear cells. AB - The ability of rabbit and monkey immune sera to neutralize prostaglandin E2 (PGE2) production by human monocytes stimulated with lipopolysaccharide (LPS) was examined. CD14-dependent LPS activation of PGE2 was examined under assay conditions which allowed the comparison of preimmune and immune sera. Serum obtained from rabbits immunized with formalin-fixed Porphyromonas gingivalis cells dramatically reduced the amount of PGE2 produced in response to LPS obtained from three different strains of P. gingivalis but not that from Escherichia coli or Bacteroides fragilis. In addition, a significant reduction in the mean PGE2 level was observed in the presence of sera from immunized but not control monkeys employed in a vaccine trial. Immune serum samples from five of nine immunized monkeys were able to reduce LPS-induced production of PGE2 by greater than 50% compared to that in the corresponding preimmune sera. Immune monkey serum, similar to immune rabbit serum, blocked PGE2 production in response to P. gingivalis LPS but not E. coli LPS. These data demonstrate that immunization with P. gingivalis whole cells can elicit an antibody response that is able to block the PGE2 response to LPS. Neutralization of LPS-mediated inflammatory mediator production may account in part for the observed suppression of alveolar bone loss in immunized monkeys. PMID- 9353069 TI - Invasion of intestinal epithelia in vitro by the parasitic nematode Trichinella spiralis. AB - Studies of nematode establishment in intestinal niches has been hindered by the lack of a readily manipulated in vitro assay. In this report, experiments are described wherein the larval stage of the parasitic nematode Trichinella spiralis was shown to invade epithelial cell monolayers in vitro. Larvae penetrated cells and migrated through them, leaving trails of dead cells in their wake. Cells derived from five different species were susceptible to invasion, reflecting the broad host range of T. spiralis in vivo. Epithelial cells derived from large and small intestines and kidneys were susceptible. Fibroblast and muscle cells were resistant. Larvae deposited glycoprotein antigens in the cells they invaded. Although the function of these antigens is unknown, they are targeted by rat antibodies that cause T. spiralis to be expelled from the intestine. The model system described provides the means to further investigate this process as well as the mechanisms by which this parasitic nematode establishes its intestinal niche. PMID- 9353067 TI - The cytoplasmic membrane is a primary target for the staphylocidal action of thrombin-induced platelet microbicidal protein. AB - Thrombin-induced platelet microbicidal protein (tPMP-1) is a small, cationic peptide released from rabbit platelets exposed to thrombin in vitro. tPMP-1 is microbicidal against a broad spectrum of bloodstream pathogens, including Staphylococcus aureus. Preliminary evidence suggests that tPMP-1 targets and disrupts the staphylococcal cytoplasmic membrane. However, it is not clear if the cytoplasmic membrane is a direct or indirect target of tPMP-1. Therefore, we assessed the in vitro activity of tPMP-1 versus protoplasts prepared from logarithmic-phase (LOG) or stationary-phase (STAT) cells of the genetically related S. aureus strains 19S and 19R (tPMP-1 susceptible and resistant, respectively). Protoplasts exposed to tPMP-1 (2 microg/ml) for 2 h at 37 degrees C were monitored for lysis (decrease in optical density at 420 nm) and ultrastructural alterations (by transmission electron microscopy [TEM]). Exposure to tPMP-1 resulted in substantial lysis of LOG but not STAT protoplasts of 19S, coinciding with protoplast membrane disruption observed by TEM. Thus, it appears that tPMP-1-induced membrane damage is influenced by the bacterial growth phase but is independent of the staphylococcal cell wall. In contrast to 19S, neither LOG nor STAT protoplasts of 19R were lysed by tPMP-1. tPMP-1-induced membrane damage was further characterized with anionic planar lipid bilayers subjected to various trans-negative voltages. tPMP-1 increased conductance across bilayers at 90 mV but not at -30 mV. Once initiated, a reduction in voltage from -90 to -30 mV diminished conductance magnitude but did not eliminate tPMP-1-mediated membrane permeabilization. Therefore, tPMP-1 appears to directly target the staphylococcal cytoplasmic membrane as a primary event in its mechanism of action. Specifically, tPMP-1 likely leads to staphylococcal death, at least in part by permeabilizing the bacterial membrane in a voltage-dependent manner. PMID- 9353071 TI - Role of tumor necrosis factor alpha in induction of murine CD14 gene expression by lipopolysaccharide. AB - We previously demonstrated CD14 gene expression in myeloid and epithelial cells of the mouse and showed that expression of the CD14 gene in both is modulated by lipopolysaccharide (LPS). Here we test the hypothesis that the induction of CD14 in these cells is an indirect effect of LPS, one mediated by tumor necrosis factor alpha (TNF-alpha). TNF-alpha induced a transient increase in levels of CD14 in plasma with a peak at 6 to 8 h, and this increase in levels of CD14 antigen in plasma was accompanied by increased levels of CD14 mRNA in lung, liver, and kidney. Moreover, in situ hybridization studies revealed that CD14 mRNA was induced in both myeloid cells and epithelial cells, the same cells that respond to LPS. Pretreatment of mice with anti-TNF antiserum reduced the LPS mediated increase in levels of CD14 in plasma and significantly reduced the level of induction of CD14 mRNA in selected epithelial cells in the kidney and liver. The antiserum did not appear to block LPS-mediated induction in myeloid cells in the tissues examined. In C3H/HeJ mice, the epithelial response to LPS was markedly attenuated whereas the response to TNF-alpha was normal. Thus, regulation of CD14 gene expression by LPS differs in epithelial and myeloid cells, with the epithelial responses in kidney and liver being mediated, in part, by TNF-alpha. PMID- 9353070 TI - Interaction of Yersinia enterocolitica with macrophages leads to macrophage cell death through apoptosis. AB - Suppression of the host defense is one of the hallmarks of Yersinia enterocolitica infection. This enteric pathogen resists phagocytosis and interferes with macrophage functions from an extracellular localization (oxidative-burst generation and tumor necrosis factor alpha production). In this study, we investigated the fate of the Y. enterocolitica-infected macrophage. We found that murine J774A.1 macrophages and macrophages derived from human monocytes were killed by infection with Y. enterocolitica. Analysis of cellular morphology and DNA fragmentation revealed that macrophage cell death occurs through the induction of apoptosis. A total of 92% +/- 5% (mean +/- standard deviation) of murine J774A.1 macrophages and 74% +/- 6% of human monocyte-derived macrophages underwent apoptosis upon Yersinia infection after 4 and 20 h, respectively. The broad-spectrum caspase inhibitor Z-Val-Ala-DL-Asp fluoromethylketone blocked completion of the Yersinia-induced apoptotic program but not the surface exposure of phosphatidylserine as an early-stage apoptotic event. Analysis of different Yersinia mutants showed that macrophage apoptosis depends on a functional Y. enterocolitica type III protein secretion system. Apoptotic cell death of macrophages was not related to the YopE-mediated cytotoxic effect of Yersinia, since disruption of actin microfilaments by a Y. enterocolitica strain expressing a restricted repertoire of yop genes, including YopE, did not result in macrophage apoptosis. Furthermore, Yersinia-induced cytotoxic alterations in epithelial HeLa cells, which are conferred by YopE, did not lead to apoptosis. Our data demonstrate for the first time that Y. enterocolitica promotes the apoptosis of macrophages, an effect which is clearly distinct from the morphological alterations mediated by Yersinia on epithelial HeLa cells. PMID- 9353072 TI - Chlamydia pneumoniae infection induces inflammatory changes in the aortas of rabbits. AB - Chlamydia pneumoniae, a common human respiratory pathogen, has been associated with atherosclerosis in several seroepidemiological studies. Moreover, its presence in lesions of vessel walls has been demonstrated by culture, immunohistochemistry, PCR, and electron microscopy. In this study, we infected intranasally with C. pneumoniae New Zealand White rabbits which had been fed a normal diet. Reinfection was given 3 weeks later. Six of the nine reinfected animals showed inflammatory changes consisting of intimal thickening or fibroid plaques resembling atherosclerosis in 2 to 4 weeks after reinfection. One rabbit had calcified lesions. Immunohistochemistry for C. pneumoniae was strongly positive in the three older affected animals. No lesions were seen in the controls. The results suggest that C. pneumoniae infection is capable of inducing inflammatory atherosclerosis-like changes in the aortas of infected rabbits. PMID- 9353073 TI - Interaction of Neisseria meningitidis with a polarized monolayer of epithelial cells. AB - An important step in the pathogenesis of Neisseria meningitidis is the crossing of two cellular barriers, one in the nasopharynx and one in the brain. To approach the mechanisms by which this bacterium can achieve these goals, we studied the interactions between N. meningitidis and a monolayer of polarized tight junction-forming T84 cells grown on filter units. A capsulated, piliated, Opa-, and Opc- N. meningitidis strain is shown to be capable of adhering to and crossing this monolayer several orders of magnitude more efficiently than an isogenic nonpiliated derivative. This bacterial interaction does not affect the barrier function of tight junctions, as assessed by (i) the absence of modification of the transepithelial resistance, (ii) the lack of increase of [3H]inulin penetration across the monolayer, and (iii) the absence of delocalization of ZO-1, a tight junction protein. Electron microscopy studies and confocal examinations demonstrated that N. meningitidis (i) induces cytoskeletal rearrangements with actin polymerization beneath adherent bacteria, (ii) is intimately attached to the apical membrane of the cells, and (iii) can be internalized inside cells. Immunofluorescent staining with antipilus antibodies showed evidence that meningococcal piliation was dramatically reduced at later time points of bacterial cell interaction compared to the early phase of this interaction. In addition, adhesive bacteria recovered from an infected monolayer are piliated, capsulated, Opa-, and Opc-, a phenotype similar to that of the parental strain. Taken together, these data demonstrate that following pilus mediated adhesion, N. meningitidis is involved in an intimate attachment which requires a bacterial component different from Opa and Opc and that meningococci cross a monolayer of tight-junction-forming epithelial cells by using a transcellular pathway rather than a paracellular route. PMID- 9353074 TI - Lethal tuberculosis in interleukin-6-deficient mutant mice. AB - Tuberculosis is a chronic infectious disease which causes major health problems globally. Acquired resistance is mediated by T lymphocytes and executed by activated macrophages. In vitro studies have emphasized the importance of macrophage activation for mycobacterial growth inhibition. In vivo, the protective host response is focused on granulomatous lesions in which Mycobacterium tuberculosis is contained. A cellular immune response of the T helper 1 (Th1) type is considered central for control of tuberculosis. Using interleukin-6 (IL-6)-deficient mice, we here demonstrate a crucial role of this pluripotent cytokine in protection against M. tuberculosis but not against Mycobacterium bovis BCG. Infection with M. tuberculosis was lethal for the IL-6 deficient mice at inocula that were still controlled by IL-6-competent mice. Spleen cells from M. tuberculosis-infected IL-6-/- mouse mutants produced elevated levels of IL-4 and reduced levels of gamma interferon compared to the control levels. Cytofluorometric analyses of spleen cells from M. tuberculosis infected mice revealed more-profound alterations in T-cell ratios in IL-6-/- mice than in control mice. We assume that IL-6 contributes to host resistance by its proinflammatory activity and by its influence on cytokine secretion. PMID- 9353075 TI - Mycobacterial growth and sensitivity to H2O2 killing in human monocytes in vitro. AB - The intracellular growth and susceptibilities to killing by H2O2 in cultured human monocytes of a number of mycobacterial species including laboratory strains and clinical isolates of Mycobacterium tuberculosis, and Mycobacterium bovis bacillus Calmette-Guerin (BCG) and a clinical isolate of Mycobacterium avium-M. intracellulare were examined. The clinical isolate of M. avium-M. intracellulare did not replicate in freshly explanted monocytes (generation time of >400 h); BCG replicated with a generation time of 95 h, and M. tuberculosis strains CDC551, H37Rv, and H37Ra replicated with generation times of 24, 35, and 37 h, respectively, during the 4-day growth assay. When cultured in monocytes for 4 days, the mycobacteria were variably sensitive to H2O2-induced killing. A positive correlation between the generation time and percent killing of intracellular bacilli was observed. By comparison, mycobacterial strains were similarly sensitive to H2O2 treatment in cell-free culture media and in sonicated cell suspensions. Using a number of inhibitors of reactive oxygen intermediates we determined that other than catalase the inhibitors tested did not affect H2O2 induced killing of intracellular mycobacteria. Our studies suggest that the killing of mycobacteria growing in human monocytes in vitro by the addition of exogenous H2O2 is dependent on the susceptibility to a peroxide-induced killing pathway as well as on the intracellular growth rate of the mycobacteria. PMID- 9353076 TI - Helicobacter bilis-induced inflammatory bowel disease in scid mice with defined flora. AB - Helicobacter bilis has been isolated from aged inbred mice with multifocal chronic hepatitis and from scid mice with diarrhea, proliferative typhlitis, and colitis. To determine the pathogenic potential of H. bilis, we inoculated 4-week old female Tac:Icr:Ha(ICR)-scidfDF mice by intraperitoneal injection of approximately 10(8) CFU of H. bilis in phosphate-buffered saline (PBS) (n = 15) or PBS alone (n = 10) and necropsied them at 7 weeks postinfection. Sham inoculated mice had no significant gross or histopathological findings. In contrast, all 15 experimentally inoculated mice (confirmed to be H. bilis colonized by culture and PCR of cecal contents) exhibited varying degrees of inflammatory bowel disease (IBD). Proliferative typhlocolitis was characterized by focal to segmental areas of crypt hyperplasia and a predominantly histiocytic inflammatory cell infiltrate. Labeling indices for 5-bromo-2'-deoxyuridine incorporation were increased approximately 2.5-fold in the ceca and colons of H. bilis-inoculated mice. This is the first study to demonstrate experimentally that infection with H. bilis causes IBD in scid mice with defined flora. This result both confirms a pathogenic role for H. bilis in mice and provides a new model relating a specific microbial agent and IBD. PMID- 9353078 TI - Study of T-lymphocyte subsets of healthy and Mycobacterium avium subsp. paratuberculosis-infected cattle. AB - The relative contributions of T-lymphocyte subsets to host defense in cattle infected with Mycobacterium avium subsp. paratuberculosis is reported. The subsets were purified with appropriate monoclonal antibodies and a magnetic bead column separation system, and their purity was verified by flow cytometry. Biological activity of each subset, expressed as lymphoproliferation and gamma interferon (IFN-gamma) production, was measured in response to phytohemagglutinin (PHA) and an M. avium antigen preparation (A-PPD). IFN-gamma was measured by antibody capture enzyme-linked immunosorbent assay. The results showed a correlation between proliferation and IFN-gamma production in response to A-PPD but not to PHA. In response to PHA, CD4+ lymphocytes were the most prolific producers of IFN-gamma. CD8+ lymphocytes produced IFN-gamma to a lesser extent, whereas gammadelta+ T lymphocytes produced little or no IFN-gamma. Differences observed between the amount of IFN-gamma produced by CD4+ versus CD8+ cells and CD4+ versus gammadelta+ cells were significant (P < 0.01), but those between peripheral blood mononuclear cells (PBMC) and CD4+ T cells were not. Similar responses to A-PPD were observed except that PBMC produced higher levels of IFN gamma than did CD4+ T cells. These data for cattle are similar to observations made for other animal species, where CD4+ cells are the major type of T lymphocytes producing IFN-gamma. They further suggest that whatever the role gammadelta+ T cells may play in paratuberculosis, it is not likely to be mediated by IFN-gamma production. PMID- 9353077 TI - Identification of superoxide dismutase activity in Borrelia burgdorferi. AB - Infective and noninfective strains of Borrelia burgdorferi, along with Borrelia afzelii and Borrelia garinii, possessed a single iron-containing superoxide dismutase (SOD). None of the Lyme disease spirochetes tested possessed catalase or peroxidase activities. The borrelial SOD was not inducible by growth with increased oxygen concentrations and thus appeared to be produced constitutively. PMID- 9353079 TI - Plasmodium falciparum- and merozoite surface protein 1-specific antibody isotype balance in immune Senegalese adults. AB - This study shows markedly different isotype distributions of antibodies to asexual blood stages of Plasmodium falciparum and to merozoite surface protein 1 in clinically immune Senegalese adults depending on the study site. The relationships between immunoglobulin M (IgM) and IgG and between IgG3 and IgG1 antibodies differed in settings where transmission is perennial compared to settings where it is seasonal. This suggests a role for antibody class and/or subclass production and utilization in the regulation of protective immunity to such antigens. PMID- 9353080 TI - Analysis of the lytic activity of the Serpulina hyodysenteriae hemolysin. AB - The hemolysins of Serpulina hyodysenteriae are active at 27 to 40 degrees C and pH 3 to 9 and are unaffected by enzymatic inhibitors. Pore formation was demonstrated by the inhibition of hemolysis with molecules of 2.0 to 2.3 nm in diameter and the release of 86rubidium from erythrocytes without hemoglobin release after exposure to native hemolysin. PMID- 9353081 TI - Differential immunogenicity of novel Mycobacterium tuberculosis antigens derived from live and dead bacilli. AB - Mouse serum raised against killed antigen preparations of Mycobacterium tuberculosis failed to recognize most of the recombinant antigens of M. tuberculosis that were originally identified by reactivity to tuberculosis (TB) patient sera. Similar results were obtained with serum from guinea pigs immunized with live and killed mycobacteria. Antibodies raised against seven random TB patient serum-reactive antigens detected each of these antigens in the sonicate preparation. The nucleotide sequences of the genes for these seven antigens revealed that all represented hitherto unreported genes of M. tuberculosis. Our results suggest differential presentation to the host immune system of the same antigens derived from live and killed mycobacteria. PMID- 9353082 TI - Cytokine profile suggesting that murine cerebral malaria is an encephalitis. AB - Cerebral malaria (CM) remains a poorly understood and life-threatening complication of malaria caused by the parasite Plasmodium falciparum. The discovery that murine CM caused by Plasmodium berghei ANKA and human CM are both characterized by production of inflammatory cytokines, especially tumor necrosis factor alpha (TNF-alpha), led to a revival of the suggestion that P. berghei CM may have value as a model of the human disease. In this study, quantitative reverse transcription-PCR was used to measure levels of message for 18S rRNA of P. berghei and 10 cytokines in the brains, livers, and spleens of mice during the induction and course of CM. A coordinated increase in RNA of parasite and proinflammatory cytokines was observed in the brains of mice in parallel with onset of CM. Levels of message for parasite, TNF-alpha, and gamma interferon increased in the brains of mice from day 5 to death on day 7. These changes were observed only in the brain, and message for other cytokines remained near baseline levels. This demonstrated that parasite sequestration does take place in the brains of mice with CM. Histologically, CM was characterized by widespread damage to the microvasculature in the brain with focal infiltration of inflammatory cells. The pattern of cytokine production in the brain is characteristic of other murine encephalitides. PMID- 9353083 TI - Cloning, expression, and sequencing of a protease gene from Bacteroides forsythus ATCC 43037 in Escherichia coli. AB - We have isolated and characterized an N-benzoyl-Val-Gly-Arg-p-nitroanilide specific protease gene, designated prtH, from Bacteroides forsythus ATCC 43037. Nucleotide sequencing of the DNA insert from the clone (hereafter referred to as clone FST) revealed that the protease activity corresponded to an open reading frame consisting of 1,272 bp coding for a 47.8-kDa protein. When plasmid pFST was used as a probe in Southern hybridization, Sau3AI-digested chromosomal DNA of B. forsythus ATCC 43037 as well as the chromosomal DNAs of the isolated strains Ta4, TR5, and YG2 showed 0.6- and 0.8-kb hybridizing bands. The cell-free extracts of clone FST showed hemolytic activity on human blood cells. The hydrolytic activity of cell extracts of the pFST clone was inhibited by p-toluenesulfonyl-L-lysine chloromethyl ketone hydrochloride, leupeptin, N-ethylmaleimide, iodoacetic acid, iodoaceteamide, and EDTA. PMID- 9353085 TI - T-cell-rich B-cell lymphoma: what is new? What is cool? PMID- 9353084 TI - Intrapulmonary Hartmannella vermiformis: a potential niche for Legionella pneumophila replication in a murine model of legionellosis. AB - The potential role of inhaled protozoa as a niche for intrapulmonary replication of Legionella pneumophila was investigated in vivo with mutant strains of L. pneumophila which have reduced virulence for the amoeba Hartmannella vermiformis. L. pneumophila AA488 and AA502 were derived from wild-type strain AA100 after transposon mutagenesis. These mutants have reduced virulence for H. vermiformis but are fully virulent for mononuclear phagocytic cells. A/J mice, which are susceptible to replicative L. pneumophila lung infections, were inoculated intratracheally with L. pneumophila AA100, AA488, or AA502 (10[6] bacteria per mouse) or were coinoculated with one of the L. pneumophila strains (10[6] bacteria per mouse) and uninfected H. vermiformis (10[6] amoebae per mouse). The effect of coinoculation with H. vermiformis on intrapulmonary growth of each L. pneumophila strain was subsequently assessed. In agreement with our previous studies, coinoculation with H. vermiformis significantly enhanced intrapulmonary growth of the parent L. pneumophila strain (AA100). In contrast, intrapulmonary growth of L. pneumophila AA488 or AA502 was not significantly enhanced by coinoculation of mice with H. vermiformis. These studies demonstrate that L. pneumophila virulence for amoebae is required for maximal intrapulmonary growth of the bacteria in mice coinoculated with H. vermiformis and support the hypothesis that inhaled amoebae may potentiate intrapulmonary growth of L. pneumophila by providing a niche for bacterial replication. PMID- 9353086 TI - A decade has passed...the Pap smear and cervical cancer. PMID- 9353087 TI - Negligible prevalence of antibodies against Trypanosoma cruzi among blood donors in the southeastern United States. AB - Trypanosoma cruzi, a hemoflagellate, causes Chagas' disease and is endemic throughout Latin America. Increasing Latin American immigration to the United States has enhanced concern about transmission of Chagas' disease by infected donor blood. The insect vector and parasites also have been found in the southeastern United States. Autochthonous infection of several species of wild and domesticated mammals suggests that the general human population also may be at risk. To assess the prevalence of antibodies to T cruzi in humans, randomly selected donor blood was screened. Initial screening was performed by indirect hemagglutination (1:4 initial serum dilution) and at least one of three different enzyme immunoassays. All samples testing positive by at least one screening method were tested by radioimmunoprecipitation and indirect immunofluorescence supplemental methods, which were used for confirmation and calculation of specificity. Of the 6,013 serum samples evaluated, 85 tested positive by one screening method. Only 10 of the samples tested positive by more than one method. The percentages of positive screening tests are 0.05% by indirect hemagglutination and 0.06%, 0.91%, 3.97% by Abbott Laboratories (Abbott Park, Ill), Gull (Gull Laboratories, Salt Lake City, Utah), and Polychaco (Polychaco S.A.I.C., Buenos Aires, Argentina) enzyme immunoassays, respectively. All samples were negative by radioimmunoprecipitation and indirect immunofluorescence. These results suggest that although parasite and vector are found in the southeastern United States and both infect mammals, the risk of natural infection to humans in this region seems to be negligible. There was variation in positivity among different screening methods. The highest percentage of positive results was with the enzyme immunoassay, in which the binding of serum antibodies to antigens is amplified by enzymatic reactions. PMID- 9353088 TI - Is the sanctuary where Helicobacter pylori avoids antibacterial treatment intracellular? AB - The sanctuary site where Helicobacter pylori evades antimicrobial therapy is unknown, but considerable data exist about an intracellular location for H pylori. Ten H pylori-infected volunteers received standard triple antimicrobial therapy for 2 weeks. Gastric mucosal biopsy specimens were obtained with jumbo forceps on therapy days 0, 3, 14, and 42. Hematoxylin-eosin staining was used for classification of gastritis and the Genta stain for the visualization of H pylori. Immunohistochemical staining was used to detect HLA-DR antigens, human heat shock protein (HSP60), and the bacterial HSP60 antigen. Bacterial HSP60 was expressed on the mucosal surface and within epithelial cells. No such expression of human HSP60 was found, which supports a bacterial origin for the intracellular HSP60. Coexpression of bacterial HSP60 and HLA-DR was always observed, indicating an ongoing local immune response. Infection was cleared on day 14, but when examined 4 weeks after completion of therapy, Genta staining indicated that only five volunteers remained free of H pylori. However, results of immunohistochemical staining were negative at this time for only two volunteers. Disappearance of intracellular expression of bacterial HSP60 remained after therapy and correlated with the intensity of chronic inflammatory cell infiltration. These data are consistent with the intracellular localization of H pylori having a role in inflammation and as a protective strategy against extracellular antibacterial activity. PMID- 9353089 TI - Granuloma inguinale (donovanosis): an unusual cause of otitis media and mastoiditis in children. AB - Granuloma inguinale (donovanosis) is seen predominantly in adults (it rarely occurs in children) and mainly affects genital skin and mucosa. Infection occurs at other skin and mucosal sites, and hematogenous dissemination to bone also has been described. The infection responds dramatically to appropriate antibiotic treatment. We present two cases of granuloma inguinale occurring in children (8 months and 5 months of age) causing mastoiditis and external ear discharges. A temporal lobe abscess also developed in the 8-month-old child. Subsequent computed tomography scans showed marked improvement in the brain lesion after treatment. The second child had a polypoid mass in the middle ear that on biopsy showed the features of granuloma inguinale. The mother of this child had biopsy proven granuloma inguinale of the uterine cervix. These cases indicate that granuloma inguinale can be transmitted during vaginal delivery, and careful cleansing of neonates born to infected mothers is recommended. PMID- 9353090 TI - Evaluation of the cytocentrifuge Gram stain as a screening test for bacteriuria in specimens from specific patient populations. AB - To assess the usefulness of the cytocentrifuge Gram stain as a urine screening test in the clinical microbiology laboratory for the elimination of culture for screen-negative specimens, we compared the results of the cytocentrifuge Gram stain to the results of culture for 1,171 urine specimens. The data were analyzed separately for specimens from males (inpatients) and females (inpatients and outpatients), as well as for catheterized and voided specimens. Overall, the cytocentrifuge Gram stain had excellent negative predictive value (97.7%) and sensitivity (92.3%) at a culture threshold of 10(5) colony-forming units per milliliter or more. The negative predictive value and sensitivity decreased at lower culture thresholds in all populations. The negative predictive value decreased most markedly for female outpatients. Because of low positive predictive value and specificity, this test is not reliable as a sole indicator for presumptive therapy in many cases with positive results. If its limitations are recognized, the cytocentrifuge Gram stain is a useful screening test for the rapid exclusion of bacteriuria. PMID- 9353091 TI - The cost-effectiveness of cervical-vaginal rescreening. AB - Although most laboratories practice 10% manual rescreening, the cost effectiveness of this and other rescreening strategies rarely has been evaluated. Using data obtained from the medical literature, a decision model was created in which rescreening strategies were compared with nonrescreening strategies for the number of false-negative and false-positive diagnoses, cancers, life expectancy, and cost-effectiveness. The strategy of 10% rescreening with a repeated cervical vaginal smear yielded almost no gain in life expectancy compared with an equivalent strategy with no rescreening. With 100% rescreening, the gain in life expectancy was only 0.24 days per patient. A 100% rescreening strategy generally was more cost-effective than a no-rescreening strategy at costs of rescreening varying from $2 to $10 per patient. A 10% rescreening strategy has limited utility. In addition, 100% rescreening strategies are more cost-effective than nonrescreening strategies, but only if the rescreening cost is low. PMID- 9353092 TI - Thymic neuroblastoma in adults: report of three cases with special emphasis on its association with the syndrome of inappropriate secretion of antidiuretic hormone. AB - We report three cases of neuroblastoma arising within the thymus of elderly patients. All tumors consisted of primitive neuroblasts showing focal gangliocytic differentiation within nests of neuropil. All stained for neuroendocrine markers but were negative for cytokeratins and for the MIC2 gene product. One tumor was associated with the syndrome of inappropriate secretion of antidiuretic hormone, an endocrinopathy we found in three of five case reports of thymic neuroblastoma in adults. Immunohistochemical stains confirmed production of antidiuretic hormone by this tumor. One patient died of progressive disease, one patient is disease free at 18 months, and the other patient died of unrelated causes, a spectrum that reflects the variable clinical behavior others have reported. The possible histogenesis of these purely neural tumors includes malignant transformation of a mediastinal teratoma, aberrantly located sympathetic ganglia, neuroectodermal cells native to the normal thymus, and precursors of thymic epithelial cells that have differentiated along neural lines. PMID- 9353093 TI - Cytologic and cytogenetic analysis of metanephric adenoma of the kidney: a report of two cases. AB - Metanephric adenoma is a recently described renal neoplasm. Because follow-up to date has been benign, accurate diagnosis on fine-needle aspiration material may be important for appropriate clinical management. A retrospective analysis of fine-needle aspiration material from two metanephric adenomas was performed. The aspirates were cellular and composed of many small to large tightly packed clusters of cells and short papillae. Occasional tubules, rosettes, and glomeruloid-like structures were seen. The tumor cells had very scant cytoplasm; small, oval to round, uniform, overlapping nuclei with fine delicate chromatin; and minute or absent nucleoli. Rare psammoma bodies were noted. Atypia, pleomorphism, necrosis, and mitoses were absent. Cytogenetic analysis showed normal karyotypes in both cases. The cytologic differential diagnosis included Wilms' tumor, renal-cortical adenoma, papillary renal cell carcinoma, and neuroendocrin, and metastatic tumors. We conclude that metanephric adenoma has unique cytologic features that may allow distinction from other renal neoplasms on fine-needle aspiration material. In difficult cases, ancillary studies may be helpful. PMID- 9353094 TI - Cardiac troponin I for the diagnosis of acute myocardial infarction in the emergency department. AB - Cardiac troponin I (TnI) was tested in 316 consecutive patients with chest pain who were admitted to the emergency department, of whom 62 were discharged with a diagnosis of acute myocardial infarction (AMI). The TnI level was abnormal in 49 patients with AMI compared with 27 for creatine kinase (CK)-MB in the first specimen obtained at admission. All 62 patients with AMI were correctly diagnosed at admission with a combination of TnI and myoglobin testing. The overall peak performance of TnI testing in samples received within 24 hours of admission indicated high sensitivity (97%) and specificity (98%) for the diagnosis of AMI. The TnI was positive in elderly patients with myocardial injury and low CK and normal CK-MB values. These data suggest that testing for TnI could replace CK-MB and, in combination with myoglobin, could facilitate the rapid and effective triage of patients with chest pain in the emergency department. PMID- 9353095 TI - Implementation of a successful on-call system in clinical chemistry. AB - Successful practice of clinical pathology depends on a wide variety of laboratory, clinical, and managerial decisions. The skills needed to make these decisions can most effectively be learned by residents and fellows in pathology using a service-oriented on-call approach. We report our experience implementing an on-call system in the clinical chemistry laboratory at the University of Louisville Hospital (Ky). We detail the guidelines used to establish this system and the elements required for its successful implementation. The system emphasizes a laboratory-initiated approach to linking laboratory results to patient care. From inception of the program during late 1990 through 1995, the number of beeper calls (including clinician contacts) steadily increased and is currently 8 to 20 per week. The on-call system is active 24 hours per day, 7 days per week, thus representing activity on all three laboratory shifts. Types of responses were separated into administrative (12%), analytical (42%), clinical (63%), quality control or quality assurance (12%), and consultation (13%) categories. We also present 6 case reports as examples demonstrating multiple elements in these categories. In 23% of the calls, clinician contact was required and achieved by the fellow or resident on call for the laboratory. The on-call reports are documented and presented informally at weekly on-call report sessions. Emphasis is placed on learning and refinement of investigative skills needed to function as an effective laboratory director. Educational emphasis for the medical staff is in establishing awareness of the presence of the laboratory as an important interactive component of patient care. In addition, we found this program to be beneficial to the hospital and to the department of pathology in fulfilling its clinical service and teaching missions. Our experience may be helpful to other institutions establishing such a program. PMID- 9353096 TI - Clinically important intermethod differences for physiologically abnormal ionized magnesium results. AB - We compared physiologically abnormal low and high ionized magnesium (iMg) results determined with the AVL 988-4 (AVL, Graz, Austria) and Nova CRT (Nova Biomedical, Waltham, Mass) ion-selective electrodes (ISEs) in serum samples from randomly selected patients. A result of < 0.39 mmol/L with either ISE constituted the low magnesium group and of > or = 0.65 mmol/L the high magnesium group. Within each group we found significant differences between the iMg results. Major intermethod differences were found for samples with physiologically normal total magnesium concentration: most of the samples in the low magnesium group (83%) had abnormally low results with the Nova ISE, whereas most of the results with the AVL ISE (83%) were normal. In contrast, all results with the AVL ISE for the high magnesium group were abnormally high, but 67% of the results with the Nova ISE were normal. The agreement for the clinical interpretation of iMg results based on the reference interval for each method was only 32%. The differences in iMg results between the two analyzers must be resolved before using the iMg test as measured with ISE for patient care. PMID- 9353097 TI - Bone marrow involvement in T-cell-rich B-cell lymphoma. AB - We describe the histologic and immunohistochemical findings in specimens from bone marrow (BM) biopsies performed for staging purposes in 13 patients with a previous tissue-based diagnosis of T-cell-rich B-cell lymphoma (TCRBCL). Bone marrow involvement was found in 8 (62%) of 13 cases and was often paratrabecular. The histologic appearance was not pathognomonic of TCRBCL, with the differential diagnosis including Hodgkin's disease and peripheral T-cell lymphoma. The infiltrates typically had a pale low-power appearance (due to histiocytic infiltration, relative hypocellularity, or both) that, in conjunction with the presence of a polymorphous infiltrate of scattered large atypical cells amid a mixed infiltrate of small lymphocytes and histiocytes, was suggestive of Hodgkin's disease. Immunohistochemistry revealed CD20 reactivity of the large atypical cells with the absence of CD15 and CD30 reactivity, supporting the diagnosis of TCRBCL. A prominent small T-cell infiltrate accompanying the large atypical cells was observed in all positive BM biopsy specimens. The increased incidence of BM involvement in TCRBCL is significantly higher than that found in de novo B-cell diffuse large cell lymphoma, suggesting a possible biologic difference between the two entities. Our cases share some similar clinicopathologic features with histiocyte-rich B-cell lymphoma and with diffuse lymphocyte-predominant Hodgkin's disease, paragranuloma type. We discuss the possible relationship to these two entities. PMID- 9353098 TI - The volume of blood shed during the bleeding time correlates with the peripheral venous hematocrit. AB - The relation among the bleeding time, the peripheral venous hematocrit, and the amount of blood shed at the template bleeding time site has not been previously defined. We studied this relation in 227 persons: 26 were patients with idiopathic thrombocytopenic purpura (ITP), 137 were patients with a variety of other bleeding disorders, and 64 were healthy subjects. The bleeding time (mean +/- SD) for the healthy group was 7.1 +/- 1.2 minutes, and the amount of shed blood was 136.4 +/- 47.2 microL; in patients with ITP the bleeding time was 14.0 +/- 4.1 minutes and the shed blood was 508.1 +/- 387 microL; and in the group with other bleeding disorders, the mean bleeding time was 9.0 +/- 3.5 minutes, and the amount of shed blood was 224.7 +/- 184 microL. Bleeding times for all persons studied showed a significant correlation of 0.75 for the amount of shed blood on the filter paper and a significant correlation of 0.28 for the peripheral venous hematocrit. There was also a significant correlation between the bleeding time and the platelet count in patients with ITP. This study demonstrates that the volume of blood shed at the bleeding time site correlates with the peripheral venous hematocrit and emphasizes the contribution of the hematocrit to primary hemostasis in healthy subjects and patients with bleeding disorders. PMID- 9353099 TI - Altered lymphocyte antigen expressions in HIV infection: a study by quantitative flow cytometry. AB - To identify surface antigen changes that may contribute to the immune deficiency in infection with the human immunodeficiency virus (HIV), we quantified, by double-staining flow cytometry, the number of antigens of the main peripheral blood lymphocyte subsets from 30 HIV-positive persons and compared them with those of 19 HIV-negative healthy donors. Standard microbeads with different capacities to bind mouse immunoglobulins were used to convert the mean fluorescence intensity values into numbers of antigen molecules per cell, measured as antibody binding capacity. The level of expression of different lymphocyte antigens in HIV-infected patients differs from that seen in normal blood lymphocytes. Some of these surface markers are decreased, whereas others are increased, and their expression is modulated depending on the specific cell subset considered. The expression of CD3, CD4, and CD8 on T lymphocytes is significantly decreased; moreover, CD3 is down-regulated on activated and nonactivated T lymphocytes and on CD4 and CD8 cells. In contrast, the expression of CD2 on T cells is significantly increased. Natural killer cells exhibit down regulation of CD7, normal levels of CD8 and CD56, and overexpression of CD2. Our results also identified, for most of these antigens, quantitative differences in membrane expression according to different disease stages, as assessed by the CD4 T-cell count. Quantitative flow cytometry therefore may provide useful insights into the lymphocyte functional defects characterizing HIV infection. PMID- 9353100 TI - Hodgkin's disease of the esophagus. AB - A 61-year-old man with acquired immunodeficiency syndrome (AIDS) sought care because of the onset of progressive dysphagia. He was found to have a perforated, fungating esophageal mass. The combined histologic and immunologic findings were diagnostic of Hodgkin's disease, nodular sclerosis type, lymphocyte-depleted variant, arising in the esophagus. The Reed-Sternberg cells and mononuclear variants were positive for Epstein-Barr virus (EBV) latent membrane protein (LMP1) and EBV RNA. Occasional small lymphoid cells were also positive for EBV RNA. Polymerase chain reaction studies demonstrated the presence of EBV type A without deletion of the EBV LMP1 gene. Other authors have reported an increased frequency of type B EBV and deletion of the EBV LMP1 gene in cases of human immunodeficiency virus-associated Hodgkin's disease. Hodgkin's disease arising in the esophagus is rare in immunocompetent patients. However, in the presence of AIDS, Hodgkin's disease should be considered in the differential diagnosis of patients with signs or symptoms of esophageal disease. PMID- 9353101 TI - Atypical glandular cells of undetermined significance. PMID- 9353102 TI - Atypical glandular cells of undetermined significance. PMID- 9353103 TI - Beta-hydroxybutyrate and acetoacetate levels. PMID- 9353104 TI - Reimbursement and cost-effective services in cervical cytology. PMID- 9353105 TI - Reimbursement and cost-effective services in cervical cytology. PMID- 9353106 TI - Controversy flares over AIDS prevention trials in third world. PMID- 9353107 TI - UK tightens regime for animal research. PMID- 9353108 TI - Genetic testing for Alzheimer's disease 'not appropriate'. PMID- 9353109 TI - NIH pilots faster feedback for grant resubmissions. PMID- 9353111 TI - Rules needed on authorship. PMID- 9353110 TI - Recent trends in the BSE epidemic. PMID- 9353112 TI - Signal transduction. Rhomantic interludes raise blood pressure. PMID- 9353113 TI - Athletics. Mostly in the mind. PMID- 9353114 TI - Deep roots for the Neanderthals. PMID- 9353115 TI - Mixed blessings for middle-aged mothers. PMID- 9353116 TI - Mixed blessings for middle-aged mothers. PMID- 9353117 TI - Mixed blessings for middle-aged mothers. PMID- 9353118 TI - Mutation of an axonemal dynein affects left-right asymmetry in inversus viscerum mice. AB - The development of characteristic visceral asymmetries along the left-right (LR) axis in an initially bilaterally symmetrical embryo is an essential feature of vertebrate patterning. The allelic mouse mutations inversus viscerum (iv) and legless (lgl) produce LR inversion, or situs inversus, in half of live-born homozygotes. This suggests that the iv gene product drives correct LR determination, and in its absence this process is randomized. These mutations provide tools for studying the development of LR-handed asymmetry and provide mouse models of human lateralization defects. At the molecular level, the normally LR asymmetric expression patterns of nodal and lefty are randomized in iv/iv embryos, suggesting that iv functions early in the genetic hierarchy of LR specification. Here we report the positional cloning of an axonemal dynein heavy chain gene, left/right-dynein (lrd), that is mutated in both lgl and iv. lrd is expressed in the node of the embryo at embryonic day 7.5, consistent with its having a role in LR development. Our findings indicate that dynein, a microtubule based motor, is involved in the determination of LR-handed asymmetry and provide insight into the early molecular mechanisms of this process. PMID- 9353119 TI - Wnt signalling required for expansion of neural crest and CNS progenitors. AB - Interactions between cells help to elaborate pattern within the vertebrate central nervous system (CNS). The genes Wnt-1 and Wnt-3a, which encode members of the Wnt family of cysteine-rich secreted signals, are coexpressed at the dorsal midline of the developing neural tube, coincident with dorsal patterning. Each signal is essential for embryonic development, Wnt-1 for midbrain patterning, and Wnt-3a for formation of the paraxial mesoderm, but the absence of a dorsal neural tube phenotype in each mutant suggests that Wnt signalling may be redundant. Here we demonstrate that in the absence of both Wnt- and Wnt-3a there is a marked deficiency in neural crest derivatives, which originate from the dorsal neural tube, and a pronounced reduction in dorsolateral neural precursors within the neural tube itself. These phenotypes do not seem to result from a disruption in the mechanisms responsible for establishing normal dorsoventral polarity. Rather, our results are consistent with a model in which local Wnt signalling regulates the expansion of dorsal neural precursors. Given the widespread expression of different Wnt genes in discrete areas of the mammalian neural tube, this may represent a general model for the action of Wnt signalling in the developing CNS. PMID- 9353120 TI - Ataxin-1 with an expanded glutamine tract alters nuclear matrix-associated structures. AB - Spinocerebellar ataxia type 1 (SCA1) is one of several neurodegenerative disorders caused by an expansion of a polyglutamine tract. It is characterized by ataxia, progressive motor deterioration, and loss of cerebellar Purkinje cells. To understand the pathogenesis of SCA1, we examined the subcellular localization of wild-type human ataxin-1 (the protein encoded by the SCA1 gene) and mutant ataxin-1 in the Purkinje cells of transgenic mice. We found that ataxin-1 localizes to the nuclei of cerebellar Purkinje cells. Normal ataxin-1 localizes to several nuclear structures approximately 0.5 microm across, whereas the expanded ataxin-1 localizes to a single approximately 2-microm structure, before the onset of ataxia. Mutant ataxin-1 localizes to a single nuclear structure in affected neurons of SCA1 patients. Similarly, COS-1 cells transfected with wild type or mutant ataxin-1 show a similar pattern of nuclear localization; with expanded ataxin-1 occurring in larger structures that are fewer in number than those of normal ataxin-1. Colocalization studies show that mutant ataxin-1 causes a specific redistribution of the nuclear matrix-associated domain containing promyelocytic leukaemia protein. Nuclear matrix preparations demonstrate that ataxin-1 associates with the nuclear matrix in Purkinje and COS cells. We therefore propose that a critical aspect of SCA1 pathogenesis involves the disruption of a nuclear matrix-associated domain. PMID- 9353121 TI - The cerebellar leucine-rich acidic nuclear protein interacts with ataxin-1. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal dominant neurodegenerative disorder characterized by ataxia, progressive motor deterioration, and loss of cerebellar Purkinje cells. SCA1 belongs to a growing group of neurodegenerative disorders caused by expansion of CAG repeats, which encode glutamine. Although the proteins containing these repeats are widely expressed, the neurodegeneration in SCA1 and other polyglutamine diseases selectively involves a few neuronal subtypes. The mechanism(s) underlying this neuronal specificity is unknown. Here we show that the cerebellar leucine-rich acidic nuclear protein (LANP) interacts with ataxin-1, the SCA1 gene product. LANP is expressed predominantly in Purkinje cells, the primary site of pathology in SCA1. The interaction between LANP and ataxin-1 is significantly stronger when the number of glutamines is increased. Immunofluorescence studies demonstrate that both LANP and ataxin-1 colocalize in nuclear matrix-associated subnuclear structures. The features of the interaction between ataxin-1 and LANP, their spatial and temporal patterns of expression, and the colocalization studies indicate that cerebellar LANP is involved in the pathogenesis of SCA1. PMID- 9353122 TI - Cutaneous lymphocyte antigen is a specialized form of PSGL-1 expressed on skin homing T cells. AB - T cells play a pathogenic role in many inflammatory and certain malignant skin diseases, including psoriasis, atopic and allergic contact dermatitis, and cutaneous T-cell lymphoma. Memory T cells that infiltrate the skin express a unique skin-homing receptor called cutaneous lymphocyte-associated antigen (CLA), a carbohydrate epitope that facilitates the targeting of T cells to inflamed skin. CLA is defined by both its reactivity with a unique monoclonal antibody, HECA-452, and its activity as a ligand for E-selectin, but the structure of the protein component of CLA has not previously been defined. Here we report that CLA is an inducible carbohydrate modification of P-selectin glycoprotein ligand-1 (PSGL-1), a known surface glycoprotein that is expressed constitutively on all human peripheral-blood T cells. Cultured peripheral-blood T cells can be differentiated into CLA-bearing cells, which bind both E-selectin and P-selectin, or CLA-negative cells, which bind P-selectin but do not bind E-selectin, suggesting that there is independent regulation of selectin-binding phenotypes. We propose that differential post-translational modification of a single cell surface receptor, PSGL-1, mediated by fucosyltransferase VII, serves as a mechanism for regulating tissue-specific homing of memory T cells. PMID- 9353123 TI - Macrophage-tropic HIV and SIV envelope proteins induce a signal through the CCR5 chemokine receptor. AB - Human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) enter target cells by forming a complex between the viral envelope protein and two cell surface membrane receptors: CD4 and a 7-span transmembrane chemokine receptor. Isolates of HIV that differ in cellular tropism use different subsets of chemokine receptors as entry cofactors: macrophage-tropic HIVs primarily use CCR5, whereas T-cell-tropic and dual-tropic isolates use CXCR4 receptors. HIV mediated signal transduction through CCR5 is not required for efficient fusion and entry of HIV in vitro. Here we show that recombinant envelope proteins from macrophage-tropic HIV and SIV induce a signal through CCR5 on CD4+ T cells and that envelope-mediated signal transduction through CCR5 induces chemotaxis of T cells. This chemotactic response may contribute to the pathogenesis of HIV in vivo by chemo-attracting activated CD4+ cells to sites of viral replication. HIV mediated signalling through CCR5 may also enhance viral replication in vivo by increasing the activation state of target cells. Alternatively, envelope-mediated CCR5 signal transduction may influence viral-associated cytopathicity or apoptosis. PMID- 9353124 TI - Polyisoprenyl phosphates in intracellular signalling. AB - In response to environmental stimuli, leukocyte membrane remodelling generates biologically active lipids that can serve as both intra- and extracellular mediators. There are several classes of lipids that can mediate inflammatory reactions. We report here on a new intracellular lipid signal that regulates oxygen-radical formation in neutrophils, a key response in microbial killing, inflammation and tissue injury. Screening of neutrophil-derived extracts rich in phosphorylated, non-saponifiable lipids revealed a potent inhibitor of superoxide anion (O2-) production. Structural analysis of biologically active fractions gave four major phosphorylated lipids: most abundant was presqualene diphosphate (PSDP). Upon activation of neutrophil receptors, PSDP and its monophosphate form, presqualene monophosphate (PSMP), undergo rapid remodelling. At submicromolar concentrations, PSDP but not PSMP inhibit O2- production by human neutrophil cell free oxidase preparations. We prepared PSDP and PSMP by total organic synthesis and matched both the physical properties and biological activity of the neutrophil-derived compounds. Our results indicate that PSDP, a recognized intermediate of cholesterol biosynthesis, is present in immune effector cells and is a potent regulator of the cellular response in host defence. PMID- 9353125 TI - Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. AB - Abnormal smooth-muscle contractility may be a major cause of disease states such as hypertension, and a smooth-muscle relaxant that modulates this process would be useful therapeutically. Smooth-muscle contraction is regulated by the cytosolic Ca2+ concentration and by the Ca2+ sensitivity of myofilaments: the former activates myosin light-chain kinase and the latter is achieved partly by inhibition of myosin phosphatase. The small GTPase Rho and its target, Rho associated kinase, participate in this latter mechanism in vitro, but their participation has not been demonstrated in intact muscles. Here we show that a pyridine derivative, Y-27632, selectively inhibits smooth-muscle contraction by inhibiting Ca2+ sensitization. We identified the Y-27632 target as a Rho associated protein kinase, p160ROCK. Y-27632 consistently suppresses Rho-induced, p160ROCK-mediated formation of stress fibres in cultured cells and dramatically corrects hypertension in several hypertensive rat models. Our findings indicate that p160ROCK-mediated Ca2+ sensitization is involved in the pathophysiology of hypertension and suggest that compounds that inhibit this process might be useful therapeutically. PMID- 9353126 TI - The Fork head transcription factor DAF-16 transduces insulin-like metabolic and longevity signals in C. elegans. AB - In mammals, insulin signalling regulates glucose transport together with the expression and activity of various metabolic enzymes. In the nematode Caenorhabditis elegans, a related pathway regulates metabolism, development and longevity. Wild-type animals enter the developmentally arrested dauer stage in response to high levels of a secreted pheromone, accumulating large amounts of fat in their intestines and hypodermis. Mutants in DAF-2 (a homologue of the mammalian insulin receptor) and AGE-1 (a homologue of the catalytic subunit of mammalian phosphatidylinositol 3-OH kinase) arrest development at the dauer stage. Moreover, animals bearing weak or temperature-sensitive mutations in daf-2 and age-1 can develop reproductively, but nevertheless show increased energy storage and longevity. Here we show that null mutations in daf-16 suppress the effects of mutations in daf-2 or age-1; lack of daf-16 bypasses the need for this insulin receptor-like signalling pathway. The principal role of DAF-2/AGE-1 signalling is thus to antagonize DAF-16. daf-16 is widely expressed and encodes three members of the Fork head family of transcription factors. The DAF-2 pathway acts synergistically with the pathway activated by a nematode TGF-beta-type signal, DAF-7, suggesting that DAF-16 cooperates with nematode SMAD proteins in regulating the transcription of key metabolic and developmental control genes. The probable human orthologues of DAF-16, FKHR and AFX, may also act downstream of insulin signalling and cooperate with TGF-beta effectors in mediating metabolic regulation. These genes may be dysregulated in diabetes. PMID- 9353127 TI - Structure of the cyclin-dependent kinase inhibitor p19Ink4d. AB - In cancer, the biochemical pathways that are dominated by the two tumour suppressor proteins, p53 and Rb, are the most frequently disrupted. Cyclin D dependent kinases phosphorylate Rb to control its activity and they are, in turn, specifically inhibited by the Ink4 family of cyclin-dependent kinase inhibitors (CDKIs) which cause arrest at the G1 phase of the cell cycle. Mutations in Rb, cyclin D1, its catalytic subunit Cdk4, and the CDKI p16Ink4a, which alter the protein or its level of expression, are all strongly implicated in cancer. This suggests that the Rb 'pathway' is of particular importance. Here we report the structure of the p19Ink4d protein, determined by NMR spectroscopy. The structure indicates that most mutations to the p16Ink4a gene, which result in loss of function, are due to incorrectly folded and/or insoluble proteins. We propose a model for the interaction of Ink4 proteins with D-type cyclin-Cdk4/6 complexes that might provide a basis for the design of therapeutics against cancer. The sequences of the Ink4 family of CDKIs are highly conserved PMID- 9353128 TI - Contribution of CNS nicotine metabolites to the neuropharmacological effects of nicotine and tobacco smoking. AB - Nicotine, the principal alkaloid in tobacco products, is generally accepted to be the active pharmacological agent responsible for CNS effects resulting from tobacco use. Arguments are presented in this commentary which take issue with this popular dogma, by providing evidence that nicotine metabolites may also be responsible for the CNS effects commonly attributed to nicotine. CNS effects attributed to nicotine include reinforcing effects, mood elevation, arousal, locomotor stimulant effects, and learning and memory enhancement. The reinforcing and locomotor stimulant effects of nicotine have been suggested to be the result of activation of CNS dopaminergic systems, and nicotine-induced modulation of dopaminergic neurotransmission has been studied in detail. Nicotine acts at a family of nicotinic receptor subtypes composed of multiple subunits; however, the exact composition of the subunits in native nicotinic receptors and the functional significance of the receptor subtype diversity are currently unknown. This nicotinic subtype diversity increases the complexity of the potential mechanisms of action of nicotine and its metabolites. Although peripheral metabolism of nicotine has been studied extensively, metabolism in the CNS has not been investigated to any great extent. Recently, studies from our laboratory have demonstrated that several nicotine metabolites are present in the CNS after acute nicotine administration. Moreover, nicotine metabolites are pharmacologically active in neurochemical and behavioral assays. Thus, CNS effects resulting from nicotine exposure may not be due solely to nicotine, but may result, at least in part, from the actions of nicotine metabolites. PMID- 9353129 TI - DNA topoisomerase II rescue by catalytic inhibitors: a new strategy to improve the antitumor selectivity of etoposide. AB - The nuclear enzyme DNA topoisomerase II (topo II) is the target of important antitumor agents such as etoposide. Recent work has classified topo II targeting drugs into either topo II poisons that act by stabilizing enzyme-DNA cleavable complexes leading to DNA breaks, or topo II catalytic inhibitors that act at stages in the catalytic cycle of the enzyme where both DNA strands are intact and, therefore, do not cause DNA breaks. Accordingly, catalytic inhibitors are known to abrogate DNA damage and cytotoxicity caused by topo II poisons. In this commentary, we have focused on the possibilities of enabling high-dose therapy with the topo II poison etoposide by protection of normal tissue with catalytic inhibitors, analogous to folinic acid rescue in high-dose methotrexate treatment. Thus, we have demonstrated recently that (+)-1,2-bis(3,5-dioxopiperazinyl-1 yl)propane (ICRF-187) enabled a 3- to 4-fold dose escalation of etoposide in mice. Two high-dose etoposide models are described, namely use of the weak base chloroquine in tumors with acidic extracellular pH and targeting of CNS tumors with protection of normal tissue by the bisdioxopiperazine ICRF-187. In conclusion, high supralethal doses of topo II poisons in combination with catalytic inhibitor protection form a new strategy to improve the antitumor selectivity of etoposide and other topo II poisons. Such an approach may be used to overcome problems with drug resistance and drug penetration. PMID- 9353130 TI - Drug metabolism in hepatocyte sandwich cultures of rats and humans. AB - Adult hepatocytes from rat and man were maintained for 2 weeks between two gel layers in a sandwich configuration to study the influence of this culture technique on the preservation of basal activities of xenobiotic-metabolizing phase I and phase II enzymes. The response of these enzyme activities to an enzyme inducer was investigated using rifampicin (RIF). Basal levels of cytochrome P-450 (CYP) isozymes were characterized by measuring ethoxyresorufin O deethylation (EROD), ethoxycoumarin O-deethylation (ECOD), and the specific oxidation of testosterone (T). In hepatocytes from untreated rats, CYP isozyme levels, including the major form CYP 2C11, increased during the first 3 days in culture. After this period of recovery, the levels of CYP 2C11, CYP 2A1, and CYP 2B1 decreased, whereas CYP 3A1 increased. In contrast to these dynamic changes, CYP activities such as CYP 1A2 and the major isozyme CYP 3A4 were largely preserved until day 9 in cultures of human hepatocytes. In measuring phase II activities, a distinct increase in glucuronosyltransferase (UDP-GT) activity toward p-nitrophenol (PNP) was found for rat and human hepatocytes over 2 weeks in culture. Sulfotransferase (ST) activity toward PNP showed an initial increase, with a maximum at day 7 and day 9 in culture, respectively, and then decreased until day 14. Glutathione S-transferase (GST) activity decreased constantly during the time of culture. Effects of the enzyme-inducing drug rifampicin on phase I and phase II enzymes were investigated using cultured human hepatocytes. Rifampicin treatment (50 micromol/L) for 7 days resulted in a 3.7-fold induction of CYP 3A4 at day 9 in culture. ECOD activity was increased sixfold and phase II ST activity increased twofold compared to the initial value at day 3. No effect of rifampicin on CYP 3A was found in cultures of rat hepatocytes. These results demonstrate that rat and human hepatocytes preserve the major forms of CYP isozymes and phase II activities and respond to inducing drugs such as rifampicin. The novel hepatocyte sandwich culture is suitable for investigating drug metabolism, drug-drug interactions and enzyme induction. PMID- 9353131 TI - Effects of membrane fatty acids on thermal and oxidative injury in the human premonocytic line U937. AB - Heat shock (HS) proteins (HSP) function as molecular chaperones and protect cells from thermal and oxidative injury. The signals leading to HSP synthesis, i.e. the "cellular thermometer(s)," are still a matter of debate. In the human premonocytic line U937, we investigated the effects of specific modification of membrane fatty acid (FA) composition by incubation with various saturated and unsaturated fatty acids (UFA) on the HS response and on hydrogen peroxide (H2O2) induced cell death. FA readily incorporated into U937 cell membranes. UFA did not modulate the HS response but potentiated H2O2-mediated damage, while pre-exposure to HS protected the UFA-treated cells from this increased H2O2 toxicity. PMID- 9353132 TI - Differential effects of tyrosine kinase inhibitors and an inhibitor of the mitogen-activated protein kinase cascade on degranulation and superoxide production of human neutrophil granulocytes. AB - The effects of two different tyrosine kinase inhibitors (genistein and erbstatin analog) and an inhibitor (2'-amino-3'-methoxyflavone; PD98059) of the mitogen activated protein (MAP) kinase kinase on the primary granule exocytosis and superoxide (O2.-) production of human neutrophil granulocytes were compared. The effector responses induced by stimulation of the chemotactic receptors by formyl methionyl-leucyl-phenylalanine and platelet-activating factor were blocked both by genistein and erbstatin analog. In contrast, degranulation and O2.- production triggered by the activation of protein kinase C with phorbol-12-myristate-13 acetate were reduced by erbstatin analog but not by genistein. This inhibitory pattern was observed in both effector responses, but the sensitivity of O2.- production toward tyrosine kinase inhibition was markedly higher than that of degranulation. PD98059 caused no considerable effect on any of the above responses. The data presented indicate that tyrosine kinases are involved not only in the respiratory burst but also in the organization of the degranulation response of neutrophil granulocytes. It is suggested that several tyrosine kinases of different inhibitor sensitivity may participate in the transduction of extracellular signals. However, activation of the MAP kinase cascade does not appear to be involved in either of the investigated biological responses of the neutrophils. PMID- 9353134 TI - Effects of troglitazone and metformin on glucose and lipid metabolism: alterations of two distinct molecular pathways. AB - Troglitazone and metformin are antidiabetic agents that belong to the thiazolidinedione and biguanide classes of drugs, respectively. To evaluate how these drugs influence fuel utilization, we compared their effects on several pathways regulating carbohydrate and lipid metabolism in vitro. Both drugs stimulated glucose transport and utilization in C3H10T1/2 cells, a cell line capable of differentiating into adipocytes when treated with thiazolidinediones. However, we observed that these drugs had a number of different in vitro effects. Unlike metformin, troglitazone stimulated beta3-adrenergic receptor-mediated lipolysis, lipogenesis, and transcriptional activity of the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma). Further, by using a mitochondrial-specific fluorescent dye, we found troglitazone to be more effective than metformin at increasing mitochondrial mass. In contrast to troglitazone, metformin was more effective at increasing mitochondrial fatty acid beta-oxidation, peroxisomal fatty acid beta-oxidation, and anaerobic respiration (i.e. lactate production). Additionally, metformin stimulated and troglitazone inhibited both aerobic respiration and basal lipolysis. Insulin enhanced the effects of troglitazone, but not those of metformin, on these cells. Taken together, the data show that troglitazone and metformin affect two distinct metabolic pathways: one that is anabolic (i.e. troglitazone) and the other that is catabolic (i.e. metformin). Further, these observations suggest that the metabolic activity of mitochondria may be lower in cells treated with troglitazone than with metformin. PMID- 9353133 TI - P-glycoprotein-independent decrease in drug accumulation by phorbol ester treatment of tumor cells. AB - The effect of a change in the phosphorylation state of the drug transporter P glycoprotein (P-gp) on its drug transport activity was studied for the substrates daunorubicin (DNR), etoposide (VP-16), and calcein acetoxymethyl ester (Cal-AM). Phorbol ester (PMA), added to stimulate phosphorylation of P-gp by protein kinase C (PKC), caused a decrease in the cellular accumulation of DNR and VP-16, both in multidrug-resistant (MDR) P-gp-overexpressing cells and in wild-type cells. Since treatment of cells with kinase inhibitor staurosporine (ST) reversed this effect of PMA and the non-PKC-stimulating phorbol ester 4alpha-phorbol, 12,13 didecanoate (4alphaPDD) did not result in a decreased DNR accumulation, we conclude that this effect is the result of kinase activity. The concentration dependence of the inhibition of P-gp by verapamil (Vp) was not influenced by PMA. Accumulation of the P-gp substrate Cal-AM was not influenced by PMA in wild-type cells. Therefore, Cal-AM was used to study the effect of PMA-induced phosphorylation of P-gp on its transport activity. Activation of PKC with PMA or inhibition of protein phosphatase 1/2A (PP1/PP2A) with okadaic acid (OA) did not affect the accumulation of Cal-AM in the MDR cells or wild-type cells. The kinase inhibitor ST increased the Cal-AM accumulation only in the MDR cells. Neither stimulating PKC with PMA nor inhibiting PP1/PP2A with OA led to a decreased inhibition of P-gp by ST, indicating that ST inhibits P-gp directly. From these experiments, we conclude that PKC and PP1/PP2A activity do not regulate the drug transport activity of P-gp. However, these studies provide evidence that PMA induced PKC activity decreases cellular drug accumulation in a P-gp-independent manner. PMID- 9353136 TI - Curcumin inhibition of Dermatophagoides farinea-induced interleukin-5 (IL-5) and granulocyte macrophage-colony stimulating factor (GM-CSF) production by lymphocytes from bronchial asthmatics. AB - Curcumin, a dietary pigment responsible for the yellow color of curry, has been used for the treatment of inflammatory diseases and exhibits a variety of pharmacological effects such as anti-inflammatory, anti-tumor, anti-oxidant, and anti-viral activity. However, it has not been determined whether the effect of curcumin on the production of cytokine affects eosinophil functions and IgE synthesis. In the present study, we examined the effect of curcumin on the production of interleukin (IL)-2, IL-5, granulocyte macrophage-colony stimulating factor (GM-CSF), and IL-4 by lymphocytes from atopic asthmatics in response to house dust mites (Dermatophagoides farinea: Df) in order to clarify a potential application for allergic diseases. Curcumin inhibited Df-induced lymphocyte proliferation and production of IL-2. Exogenous IL-2 reconstituted the proliferative responsiveness of lymphocytes to Df in the presence of curcumin. Furthermore, curcumin inhibited IL-5, GM-CSF, and IL-4 production in a concentration-dependent manner. These results indicate that curcumin may have a potential effect on controlling allergic diseases through inhibiting the production of cytokines affecting eosinophil function and IgE synthesis. PMID- 9353135 TI - Mobilization of Ca2+ from intracellular stores in transfected neuro2a cells by activation of multiple opioid receptor subtypes. AB - In neuronal cell lines, activation of opioid receptors has been shown to mobilize intracellular Ca2+ stores. In this report, we describe the excitatory actions of opioid agonists on murine neuroblastoma neuro2a cells stably expressing either delta, mu, or kappa opioid receptors. Fura-2-based digital imaging was used to record opioid-induced increases in intracellular Ca2+ concentration ([Ca2+]i). Repeated challenges of delta, mu, or kappa opioid receptor expressing cells with 100 nM [D-Ala2,D-Leu5]-enkephalin (DADLE), [D-Ala2,N-Me-Phe4,Gly-ol]-enkephalin (DAMGO), or trans-(+/-)-3,4-dichloro N-methyl-N-(2-[1-pyrollidinyl] cyclohexyl) benzene acetamide (U-50488H), respectively, elicited reproducible Ca2+ responses. Non-transfected neuro2a cells did not respond to opioid agonists. Removal of extracellular Ca2+ from the bath prior to and during agonist challenge did not affect significantly the agonist-evoked increase in [Ca2+]i, indicating that the response resulted from the release of Ca2+ from intracellular stores. Naloxone reversibly inhibited responses in all three cell lines, confirming that they were mediated by opioid receptors. Expression of cloned opioid receptors in neuro2a cells, coupled with digital [Ca2+]i imaging, provides a model system for the study of opioid receptors and opioid-activated signaling processes. The fact that all three receptors coupled to the same intracellular signaling mechanism suggests that the primary functional difference between opioid responses in vivo results from their selective localization. PMID- 9353137 TI - Activation of neurotensin receptors and purinoceptors in human colonic adenocarcinoma cells detected with the microphysiometer. AB - Activation of endogenous neurotensin (NT) receptors and P2-purinoceptors expressed by human colonic adenocarcinoma HT-29 cells increased extracellular acidification rates that were detected in the microphysiometer. NT (pGlu-Leu-Tyr Glu-Asn-Lys-Pro-Arg-Arg-Pro-Tyr-Ile-Leu), NT[8-13] (Arg-Arg-Pro-Tyr-Ile-Leu), NT[9-13] (Arg-Pro-Tyr-Ile-Leu), and NT1 (N alpha methyl-Arg-Lys-Pro-Trp-Tle-Leu [Tle = tert-leucine]) were full agonists, whereas XL 775 (N-[N-[2-[3-[[6-amino-1 oxo-2-[[(phenylmethoxy)carbonyl]-amino]hex yl]amino]phenyl]-3-(4-hydroxyphenyl)-1 oxo-2-propenyl]-L-isoleucyl]-L-le ucine) was a partial agonist for activating NT receptors expressed by HT-29 cells. Desensitization induced by NT was rapid and monophasic with 85% of the initial response lost by a 30-s exposure. Once initiated, the rate and extent of desensitization were similar for different concentrations of a given agonist, for agonists of different potencies, and for agonists of different efficacies, which suggests that desensitization may be independent of receptor occupancy or agonist efficacy. Resensitization was a much slower process, requiring 60 min before the full agonist response to NT was recovered. ATP, via P2-purinoceptors, also activated cellular acidification rates in a concentration-dependent manner. ATP induced a biphasic desensitization of purinoceptors with a loss of ca. 50% of the initial stimulation detectable between 30 and 90 s of exposure to the agonist. Desensitization of NT receptors did not influence the activation of P2-purinoceptors by ATP, suggesting there was no heterologous desensitization between the two types of receptors. Superfusion with NT receptor agonists for 15 min at concentrations that did not elicit changes in extracellular acidification rates blocked, in a concentration dependent manner, the agonist response induced by 100 nM NT. This may reflect sequestration of the receptor. These results suggest that the high agonist affinity state of NT receptors may modulate receptor sequestration, whereas activation of the low agonist affinity state may be linked to cellular metabolism. Comparison of our results with published data found differences as well as similarities of NT responses among three lines of HT-29 cells. PMID- 9353138 TI - Influence of acute and chronic ethanol treatment on muscarinic responses and receptor expression in Chinese hamster ovary cells. AB - The influence of ethanol on the muscarinic receptor-mediated release of inositol phosphate from Chinese hamster ovary (CHO) cells stably transfected with one of the five subtypes of muscarinic acetylcholine receptor was determined. In CHO cells expressing M3 muscarinic receptors (CHO-M3), carbamylcholine increased muscarinic receptor-induced release of inositol phosphate by 150-350% following a 15-min incubation with an EC50 of approximately 30 microM. Maximal responses were obtained with 1 mM carbamylcholine, while responses to 10 mM carbamylcholine were somewhat less than maximal. Preincubation with atropine for 10 min inhibited the response with an IC50 of approximately 30 nM. CHO cells transfected with M1, M3, and M5 receptors displayed a similar pattern of activity; CHO cells transfected with M2 and M4, as well as untransfected cells, were unresponsive to carbamylcholine. Ethanol acutely inhibited the response of CHO-M3 cells to carbamylcholine by 15% at 18 mM and by 47% at 180 mM (the highest concentration examined). CHO-M3 cells were incubated with 50 mM ethanol for 48 hr. This treatment did not affect the number of cells or their protein content (113 pg/cell). The expression of M3 muscarinic receptors (determined using [3H]N methylscopolamine) increased from 1.34 +/- 0.23 to 1.75 +/- 0.16 pmol/mg protein (P < 0.05). In contrast, carbamylcholine-stimulated release of inositol phosphate was depressed by 40-70% in four experiments. Concentration-response analyses indicated a non-competitive inhibitory mechanism. This dissociation of muscarinic receptor expression and muscarinic signaling suggests a compensatory increase in receptor expression in response to chronic inhibition of muscarinic signaling by ethanol. PMID- 9353139 TI - Developmental changes in the constitutive and inducible expression of cytochrome P450 3A2. AB - Using a CYP3A2-specific oligonucleotide and an antipeptide antibody raised against the C terminus of CYP3A2 (VINGA) it is demonstrated that metyrapone administration to adult (12 weeks old) but not immature (3 weeks old) male Sprague Dawley rats induces the hepatic expression of CYP3A2 mRNA and protein. The constitutively expressed level of CYP3A2 protein in adult male rats is markedly lower than the levels expressed in immature rats as determined using the anti-VINGA antibody, in contrast to previous reports using antibodies that do not discriminate between CYP3A forms. Hepatic microsomal CYP3A2 protein expression, examined between 3 and 15 weeks of age, is extinguished between 9 and 12 weeks of age in contrast to immunoreactive CYP3A protein (determined using a nonselective antibody) and CYP3A-dependent androstenedione 6beta-hydroxylase activity. These data suggest that the regulation of the induction of CYP3A2 is developmentally controlled and that the major expressed adult form(s) of constitutively expressed CYP3A is not CYP3A2. PMID- 9353140 TI - Peripheral anergy and local immune hyperactivation in sarcoidosis: a paradox or birds of a feather. PMID- 9353141 TI - Clinical outcome of hypogammaglobulinaemic patients following outbreak of acute hepatitis C: 2 year follow up. AB - In 1994, an outbreak of hepatitis C virus (HCV) infection, genotype 1a, occurred in 30 hypogammaglobulinaemic patients in the UK from one batch of contaminated anti-HCV screened intravenous immunoglobulin. This study aimed to study prospectively the outcome of HCV in hypogammaglobulinaemic patients, and to assess the response to early treatment with interferon-alpha, 6 million units three times weekly for 6 months. Data were collected using standardized questionnaires. Five patients with secondary hypogammaglobulinaemia due to lymphoid malignancy were not treated and all have died of their primary malignancy. Of 25 patients with primary hypogammaglobulinaemia, one resolved HCV infection before treatment, 17 commenced on treatment, and seven declined or treatment was contra-indicated. Thirteen of 17 patients completed therapy and seven (54%) have a sustained response (normal transaminases, negative serum HCV RNA) at 6 and 12 months after treatment. Two of the 12 patients with primary hypogammaglobulinaemia, who were not treated or failed to complete treatment, have cleared the virus. Liver biopsy was performed in patients not clearing HCV and was abnormal in all. Four patients developed liver failure within 2 years, of whom three have died and one has been successfully transplanted. In conclusion, HCV can cause rapid severe liver disease in hypogammaglobulinaemic patients. Early treatment with high-dose interferon-alpha results in a high clearance of HCV. PMID- 9353143 TI - In vitro measurement of cytotoxic T cell activity does not predict clinical progression in paediatric HIV disease--two case studies. AB - Cytotoxic T cells are believed to be an important immune response in HIV infection, both in the initial response to viraemia, and in controlling HIV replication and maintaining clinical stability. We report here the detailed findings in two vertically infected children, from the Edinburgh perinatal cohort. Both were clinically stable for the first 7 years of life. One had vigorous HIV-specific cytotoxic T lymphocyte (CTL) responses, and non-lytic suppression, measured in vitro, while the second had no CTL activity against HIV. Despite her HIV-specific immunity, the first child had a declining CD4 count, and a high and fluctuating viral load, whereas the second child maintained a stable CD4 count, a low viral load and had a virus which could not be cultured in peripheral blood mononuclear cells (PBMC) in vitro. The first child subsequently progressed to AIDS and has now died, while the second remains clinically well. More detailed investigations showed the clinically stable child to be heterozygous for the CCR5 receptor, and to be HLA-B49--both of which markers have been associated with slow HIV disease progression. These findings question the role of CTL in maintaining stable HIV disease, and stress the need for immunological investigations to be considered in the light of the genetic make-up of the patient. They may also reflect a different immunopathogenesis of HIV disease in children compared with adults. PMID- 9353142 TI - Impaired hepatosplenic elimination of circulating cryoglobulins in patients with essential mixed cryoglobulinaemia and hepatitis C virus (HCV) infection. AB - The pathogenic mechanisms that lead to renal deposition of the cryoprecipitable IgM rheumatoid factor-IgG complexes in essential mixed cryoglobulinaemia (EMC) are unknown. Defective removal of cryoprecipitable complexes from the circulation has been postulated in EMC-associated nephritis. To test this hypothesis, the kinetics and fate of a trace dose of 123I-radiolabelled autologous cryoglobulins were analysed in 13 patients with EMC grouped according to renal involvement. The time course of radioactivity distribution in the blood and organ uptake were measured by gamma camera scintigraphy. In blood sampled 30-300 s after injection, only a minor fraction (< 15%) of the circulating cryoglobulins bound to the erythrocytes, suggesting the elimination mechanisms are independent of binding to CR1 on erythrocytes. The overall blood disappearance curve showed a fast (< or = 1 min) and slow (> 4 h) biphasic pattern. In patients with quiescent or mild nepthritis, the liver and to a lesser extent the spleen were the major organs that mediated the rapid uptake and processing of the cryoglobulins from the circulation. In contrast, patients with active mesangiocapillary glomerulonephritis showed significantly (P < 0.001) less hepatic uptake, low liver-to-precordium ratio, and slower processing of cryoglobulins, prolonged liver mean transit time, than quiescent patients or mild nephritis patients. To elucidate the role and influence of HCV infection in the pathogenesis of EMC nephritis, sera and cryoglobulins from all patients were assayed for HCV. None of the control group cases without nephritis showed any evidence of HCV-RNA in serum or cryoglobulin pellet. In contrast, all 10 EMC-nephritis patients' sera, and eight corresponding cryoglobulin pellets contained HCV-RNA. Collectively, these findings suggest an impaired reticuloendothelial system removal of IgM-IgG-HCV complexes may underlie their renal deposition. PMID- 9353144 TI - Altered sialylation of alveolar macrophages in HIV-1-infected individuals. AB - In previous studies, we have demonstrated that O-glycans at the surface of HIV-1 infected cell lines were hyposialylated. Moreover, we and others have shown that HIV+ individuals produced autoantibodies that react with hyposialylated CD43, on T cell lines. Since the autoantigen responsible for this abnormal immune response was not easily found in the peripheral blood cells of corresponding patients, we searched for its possible presence in other sites. Using fluorescence staining of alveolar macrophages with various lectins, we show that the binding of the PNA lectin specific for asialo O-glycans is much more efficient on cells from HIV-1 infected individuals. Moreover, the degree of reactivity of PNA is correlated with the clinical stage of the illness. PMID- 9353145 TI - Cell-mediated immune responses to mycobacterial antigens in patients with pulmonary tuberculosis and HIV infection. AB - Lymphocyte proliferation and cytokine responses induced by a panel of mycobacterial antigens were compared in Portuguese donors with pulmonary tuberculosis (TB) with or without HIV co-infection, HIV+ patients and healthy Mantoux-positive controls. Control donors showed stronger proliferative responses than any of the patient groups, with secreted antigens (Mycobacterium tuberculosis (Mtb) 30 kD and short-term culture filtrate proteins (ST-CFP)), purified protein derivative (PPD) and Mtb H37Rv Sonicate (MtbS) inducing the strongest proliferation. Patients with pulmonary TB showed lower proliferation to PPD or to the 30-kD antigen. Responses to all the antigens (PPD, ST-CFP, MtbS, 70 kD, 65 kD, 38 kD, 30 kD and 10 kD) were higher in TB/HIV patients with CD4 counts > or = 200 CD4+ T cells/mm3 compared with HIV alone (CD4 > or = 200 T cells/mm3), but were lost in both TB/HIV and HIV patients when CD4 counts fell below 200 T cells/mm3. Measurements of interferon-gamma (IFN-gamma) in culture supernatants revealed that PPD, 30 kD, MtbS and ST-CFP induced the strongest Th1 response. Analysis of mRNA for IFN-gamma, IL-4 and IL-10 confirmed that IFN-gamma production was maintained in patients with pulmonary TB without any concomitant increase in IL-4 or IL-10 mRNA expression, although expression of IL-10 mRNA was increased if HIV infection was present. These results reveal that IFN-gamma production is retained in pulmonary TB patients to a broad range of mycobacterial antigens, and that no switch to IL-4 production is seen even with HIV infection. Secreted antigens, and in particular ST-CFP, were the best inducers of IFN-gamma secretion, confirming their role in protective responses to Mtb. PMID- 9353146 TI - Antibodies against dengue virus E protein peptide bind to human plasminogen and inhibit plasmin activity. AB - Both mice and rabbits immunized with dengue virus E protein peptide spanning amino acids 100-119 (D4E) produced antibodies that reacted not only with the D4E peptide itself but also with human plasminogen, as shown by ELISA and Western blot. Sera from dengue virus-hyperimmunized mice and dengue patients also contained antibodies against D4E and plasminogen. Furthermore, such sera all contained plasmin inhibitory activity. Using affinity-purified anti-D4E antibodies and free D4E peptide for competitive inhibition, we demonstrated that the inhibition of plasmin activity was due to anti-D4E antibodies rather than other substances in the sera. Taken together, these results suggest dengue virus E protein amino acids 100-119 are a cross-reactive immunogenic region, and antibodies against this region may interfere with human fibrinolysis. PMID- 9353147 TI - Anti-neutrophil cytoplasmic antibody (ANCA) in malaria is directed against cathepsin G. AB - Autoantibodies of diverse specificities are detected in sera of patients with acute malaria. The clinical relevance of these autoantibodies is not clear, though there are reports associating some autoantibodies with specific disease manifestations. We have investigated the occurrence of ANCA in the sera of 93 patients during episodes of acute malaria. Sera were tested by indirect immunofluorescence (IIF) and by ELISA for antibodies to neutrophil cytoplasmic components proteinase 3 (PR3), myeloperoxidase (MPO), cathepsin G (CG), human leucocyte elastase (HLE), and lactoferrin (LF). Forty-seven sera samples (50.5%) were positive by IIF, all except one with the atypical ANCA pattern (a-ANCA). When screened by ELISA, anti-CG antibodies were detected in 52 samples (56%), while anti-PR3 and anti-MPO antibodies were detected in three and one samples, respectively. Antibody binding to HLE and LF was not significant. Anti-CG antibodies were detected in 93% of the IIF-positive sera. A combination of anti CG and anti-PR3 antibodies was noted in three samples. Our study demonstrates the presence of ANCA in sera from patients with acute malaria, almost all with the a ANCA pattern on IIF. The antibody specificity, noted for the first time in our study, appears to be predominantly directed against CG. The significance of CG and CG-ANCA in the pathogenesis and clinical manifestations of malaria has yet to be elucidated. PMID- 9353149 TI - Characterization of murine monoclonal antibodies against the Ro52 autoantigen. AB - Immunization of BALB/c mice with purified recombinant human Ro52 protein resulted in three anti-Ro52 MoAbs termed 2E7, 4C6 and 4F11. All anti-Ro52 MoAbs specifically reacted with recombinant human Ro52 protein, and also with Ro52 protein in total extracts of all human cell lines analysed, including the epithelial cell line HeLa, the B cell line Raji, the bladder carcinoma cell line RT112, and a fibroblast cell line derived from patients with xeroderma pigmentosum. The anti-Ro52 MoAbs were able to immunoprecipitate the recombinant human Ro52 protein expressed in wheat germ extract, but failed to precipitate hY RNAs from cell extracts. The staining pattern of the MoAbs strongly differed between the RT112 cells and the fibroblast cell line. RT112 cells displayed an intense cytoplasmic staining and in addition distinct fine nuclear speckles. In contrast, in the fibroblast cell line no cytoplasmic staining but only staining of distinct nuclear speckles was observed. Using deletion mutants of Ro52 the epitopes recognized by the anti-Ro52 MoAbs 2E7, 4C6 and 4F11 were partially mapped. All three MoAbs appeared to recognize distinct epitopes, that are located in the regions of Ro52 bordered by amino acids 136-164, 208-363 and 136-190, respectively. These MoAbs can be of great use in studying the cellular processes in which the Ro52 protein is involved. PMID- 9353148 TI - Local therapy with soluble complement receptor 1 (sCR1) suppresses inflammation in rat mono-articular arthritis. AB - Complement activation has been implicated in the pathogenesis of human rheumatoid arthritis. We sought to determine whether inhibition of complement (C) using sCR1 could influence the development and progression of antigen arthritis in the rat, a recognized model of human chronic synovitis. The effect of C inhibition, systemically and locally, on three different stages of disease was examined: (i) prophylaxis, (ii) treatment of established inflammation, and (iii) prevention of antigen-induced flares of disease. Arthritis was assessed by knee swelling and by histological examination. Our results show that intra-articular injection of sCR1 prior to disease onset reduced joint swelling and development of arthritis, whereas systemic administration was ineffective. Treatment of established arthritis with intraarticular sCR1 3 days after disease onset caused a transient reduction in swelling, but treatment 7 days after disease onset had no effect on disease. An intra-articular dose of sCR1 given at the time of disease flares had a small, yet significant effect on knee swelling. We conclude that complement activation is important in the initiation and maintenance of inflammation in antigen arthritis. The potent effect of local C inhibition suggests that C biosynthesis and activation within the joint contributes to inflammation in this model of arthritis. PMID- 9353150 TI - Anti-CD2 (OX34) MoAb treatment of adjuvant arthritic rats: attenuation of established arthritis, selective depletion of CD4+ T cells, and CD2 down modulation. AB - Anti-CD2 MoAbs have previously been shown to induce tolerance and to block B cell differentiation, T cell and monocyte activation. Since these immune functions are important in joint inflammation, we asked whether administration of the anti-CD2 MoAb OX34 has a beneficial effect on established rat adjuvant arthritis, a model of human rheumatoid arthritis, and how it affects CD2-bearing leucocyte subsets. Female Lewis rats with established adjuvant arthritis received a total of 5 mg OX34 or isotype-matched control MoAb starting on day 15 after adjuvant injection. Weight and arthritis score (AS) were measured in a blinded fashion. Peripheral blood cells were analysed for numbers of leucocyte subsets at various time points. Animals were killed on day 30 and lymphatic organs were processed for immunohistology. Clinically, OX34 treatment led to increased body weight and reduced AS. Although OX34 binds to CD4+ and CD8+ T cells in a comparable fashion, OX34 treatment reduced CD4+ T cells, but not CD8+ T cells. Among CD4+ T cells CD45RC+ ('naive') T cells virtually disappeared; CD45RC- ('recently activated') T cells were slightly reduced. A reduction of CD4+ T cells was also found in the lung, liver, bone marrow, spleen and lymph nodes. Down-modulation of the CD2 molecule by OX34, again, affected CD4+ T cells, suggesting a specific signal for CD4+ but not CD8+ T cells. In conclusion, the anti-CD2 MoAb OX34 attenuates established rat adjuvant arthritis. In spite of similar binding to CD4+ and CD8+ T cells, OX34 depletes only CD4+ T cells and down-modulates the CD2 molecule on these cells. These results suggest a therapeutic benefit from CD2-directed therapy for chronic types of arthritis. PMID- 9353151 TI - Experimental immunization with anti-rheumatic bacterial extract OM-89 induces T cell responses to heat shock protein (hsp)60 and hsp70; modulation of peripheral immunological tolerance as its possible mode of action in the treatment of rheumatoid arthritis (RA). AB - OM-89 is a bacterial (Escherichia coli) extract used for oral administration in the treatment of RA. Given the evidence that immunity to bacterial heat shock antigens plays a critical role in the immunomodulation of arthritis and possibly inflammation in general, the purpose of the present studies was to evaluate the presence and immunogenicity of hsp in OM-89. Furthermore, we studied the effects of OM-89 in an experimental arthritis, where hsp are known to have a critical significance in disease development. In rats immunization with OM-89 was found to lead to proliferative T cell responses to hsp60 and hsp70 of both E. coli and mycobacterial origin. Conversely, immunization with hsp antigens was also found to induce T cell reactivity specific for OM-89. Based on this and the antigen specificity analysis of specific T cell lines, hsp70(DnaK) turned out to be one of the major immunogenic constituents of OM-89. Parenteral immunization with OM 89 was found to reduce resistance to adjuvant arthritis (AA), whereas oral administration was found to protect against AA. Given the arthritis-inhibitory effect of oral OM-89 in AA, it is possible that peripheral tolerance is induced at the level of regulatory T cells with specificity for hsp. This may also constitute a mode of action for OM-89 as an arthritis-suppressive oral drug. PMID- 9353152 TI - Defective de novo thymocyte maturation in cyclosporin A (CsA)-induced autoimmunity: expression of costimulatory and activation molecules. AB - Lethally x-irradiated Lewis rats, reconstituted with syngeneic bone marrow and transiently treated with CsA for 4 weeks, will develop an autoimmune disease about 2-3 weeks after cessation of CsA therapy. CsA-induced autoimmunity is a thymus-dependent and T cell-mediated autoimmune disease. CsA is thought to generate autoreactive T cells by interference with negative selection in the thymus; x-irradiation is required to eliminate the peripheral autoregulatory T cell circuit. In this study we re-evaluate the effect of CsA on thymic atrophy and thymocyte maturation. Subsequently we examine the expression of costimulatory and activation molecules (CD2, CD5, CD11a, CD11b, CD25, CD28, CD43, CD54, OX-40, RT-1A, RT-1B and RT-1D) during distinct maturational stages in order to detect possible clues to the observed effects of CsA on thymocyte maturation and selection. The results revealed that CsA blocks maturation of double-positive TCR(int) to double-positive TCR(high) thymocytes and preferentially inhibits the development of mature CD4 single-positive thymocytes. Furthermore, CsA administration resulted in a reduced expression of the costimulatory CD2 molecule. Although it is a matter of debate whether this defective CD2 expression is involved in the aberrant maturation and selection of thymocytes, it is speculated that reduced costimulation via CD2 may influence differentiation into distinct T cell subsets. PMID- 9353153 TI - Mercury-induced renal immune complex deposits in young (NZB x NZW)F1 mice: characterization of antibodies/autoantibodies. AB - It is well demonstrated that mercury induces a systemic autoimmune disease in susceptible mouse strains. One of the major characteristics of mercury-induced autoimmune disease in mice is the development of renal immune complex deposits. We have previously shown that continual injection of mercury into young autoimmune prone (NZB x NZW)F1 mice induced an increase in antibody/autoantibody production as well as development of early renal immune complex deposits. In the present study, we characterized the isotype, the specificity and the possible pathogenicity of deposited immunoglobulins in the kidneys of mercury-injected (NZB x NZW)F1 hybrids. We found that young (NZB x NZW)F1 mice injected with mercuric chloride (HgCl2) for 6 weeks developed intense antibody formation of all immunoglobulin isotypes (except for IgG2b) as well as high levels of granular deposits of IgM, IgG1, IgG2a and IgG3 antibodies in the renal mesangium. Increased levels of the same antibody isotypes were also found in the kidney eluate of mercury- but not saline-injected mice. The dominant antibody in the kidney eluate of mercury-injected mice was of IgG1 isotype and found to be directed against double-stranded DNA, collagen, cardiolipin, phosphatidylethanolamine, and the hapten trinitrophenol, but not against nucleolar antigens. Further studies demonstrated that mercury-induced renal immune complex deposits in young (NZB x NZW)F1 mice did not lead to a severe kidney injury. Thus, in response to mercury, young (NZB x NZW)F1 mice develop renal immunoglobulin deposits with an isotype and specificity pattern correlating with that seen in the spleen and in the serum. PMID- 9353154 TI - An assessment of peripheral immunity in patients with sarcoidosis using measurements of serum vitamin D3, cytokines and soluble CD23. AB - The aetiology of the peripheral anergy in sarcoidosis is unclear. To investigate this further we measured the serum levels of several factors important in different aspects of immune regulation to obtain a profile of those factors which promote and inhibit immune activation in sarcoidosis. Thirty-seven patients with sarcoidosis and 20 healthy controls of similar sex and age comprised the study group. Serum IL-10, interferon-gamma (IFN-gamma), soluble CD23 (sCD23), IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-1beta and tumour necrosis factor-alpha (TNF-alpha) were measured using in-house ELISAs. Vitamin D3 was measured using a radioreceptor assay. Serum levels of sCD23 and IL-10 were significantly elevated in patients with sarcoidosis relative to controls (median 13.9 versus 9.5 arbitrary units/ml, P<0.01 for sCD23, and 9.6 versus 5.0 pg/ml, P<0.04 for IL-10). Regardless of steroid therapy or disease activity, serum levels of IFN-gamma, TNF-alpha, IL-1beta, GM-CSF and IL-8 were no different in patients with sarcoidosis and controls. Vitamin D3 levels were significantly higher in patients with sarcoidosis versus normal controls (medians 78.0 versus 56.0, P<0.001), active sarcoidosis (n = 20) versus inactive disease (n = 17) (medians 81.5 versus 66.0, P<0.03) and active sarcoidosis versus controls (medians 81.5 versus 56.0, P<0.0002). The levels were no different between patients with inactive sarcoidosis and controls. We suggest that IL-10 and vitamin D3 may contribute to the peripheral anergy in sarcoidosis. The elevated serum sCD23 suggests an increase in peripheral humoral immunity. Consistent with a quiescent peripheral immune system, factors capable of monocyte/macrophage activation (TNF-alpha, IFN-gamma, GM-CSF and IL-8) were not elevated in the peripheral circulation. PMID- 9353155 TI - CTLA-4 gene polymorphism confers susceptibility to insulin-dependent diabetes mellitus (IDDM) independently from age and from other genetic or immune disease markers. The Belgian Diabetes Registry. AB - Apart from genes in the HLA complex (IDDM1) and the variable number of tandem repeats in the 5' region of the insulin gene (INS VNTR, IDDM2), several other loci have been proposed to contribute to IDDM susceptibility. Recently, linkage and association have been shown between the cytotoxic T lymphocyte-associated protein 4 (CTLA-4) gene on chromosome 2q and IDDM. In a registry-based group of 525 recent-onset IDDM patients <40 years old we investigated the possible interactions of a CTLA-4 gene A-to-G transition polymorphism with age at clinical disease onset and with the presence or absence of established genetic (HLA-DQ, INS VNTR) and immune disease markers (autoantibodies against islet cell cytoplasm (ICA); insulin (IAA); glutamate decarboxylase (GAD65-Ab); IA-2 protein tyrosine phosphatase (IA-2-Ab)) determined within the first week of insulin treatment. In new-onset IDDM patients. G-allele-containing CTLA-4 genotypes (relative risk (RR)= 1.5; 95% confidence interval (CI) = 1.2-2.0; P < 0.005) were not preferentially associated with age at clinical presentation or with the presence of other genetic (HLA-DR3 or DR4 alleles; HLA-DQA1*0301-DQB1*0302 and/or DQA1*0501-DQB1*0201 risk haplotypes; INS VNTR I/I risk genotype) or immune (ICA, IAA, IA-2-Ab, GAD65-Ab) markers of diabetes. For 151 patients, thyrogastric autoantibodies (anti-thyroid peroxidase, anti-thyroid-stimulating hormone (TSH) receptor, anti-parietal cell, anti-intrinsic factor) were determined, but association between CTLA-4 risk genotypes and markers of polyendocrine autoimmunity could not be demonstrated before or after stratification for HLA- or INS-linked risk. In conclusion, the presence of a G-containing CTLA-4 genotype confers a moderate but significant RR for IDDM that is independent of age and genetic or immune disease markers. PMID- 9353156 TI - Differential distribution of B7.1 (CD80) and B7.2 (CD86) costimulatory molecules on mucosal macrophage subsets in human inflammatory bowel disease (IBD). AB - The molecules B7.1 and B7.2 deliver costimulatory signals of critical importance to naive T cells, and may thus be involved in abrogation of oral tolerance in IBD. Functional disparity apparently exists among antigen-presenting cells in vivo. We wanted to examine if differential B7 expression occurs on mucosal macrophage subsets. Cryosections of bowel specimens from patients with IBD and normal controls were subjected to immunofluorescence and immunoperoxidase staining. In normal mucosa, selective subepithelial accumulation of B7.2+ cells was found. In inflamed IBD mucosa, however, subsets appeared consisting of both B7.2(hi) and B7.1(hi) cells as well as CD14(hi) macrophages. Notably, outside lymphoid aggregates the prominent fraction of recently recruited CD14(hi) macrophages comprised most (approximately 80%) of the B7.1(hi) cells, whereas most (approximately 70%) B7.2(hi) cells were identified as resident mucosal macrophages (CD14(lo) or CD14-). Differential expression of B7.1 and B7.2 on two functionally different subsets of intestinal macrophages implies separate immunoregulatory roles for the two molecules. This finding is in keeping with recent experimental data demonstrating that monocyte-derived cells are crucial for immune responses at mucosal surfaces. Preferential B7.1 up-regulation might be critical in breaking the immunological tolerance to luminal antigens in IBD, but it cannot be excluded that it is a secondary pathogenic event. PMID- 9353157 TI - IL-5 production by allergen-stimulated T cells following grass pollen immunotherapy for seasonal allergic rhinitis. AB - Grass pollen immunotherapy for the treatment of seasonal allergic rhinitis ('summer hayfever') results in improvement in symptoms, a reduction in the early and late phase responses to allergen provocation and decreased tissue eosinophilia. Immunotherapy may act by altering the pattern of cytokine production by allergen-specific T cells from a 'Th2-type' (IL-4 and IL-5) profile to a 'Th1-type' (interferon-gamma (IFN-gamma)) profile. We set out to determine whether clinical improvement following specific allergen immunotherapy is accompanied by reduced production of the pro-eosinophilic and archetypal 'Th2 type' cytokine, IL-5. Peripheral blood mononuclear cells (PBMC) were isolated from (i) 13 patients who had received 6 or 7 years' continuous conventional immunotherapy with timothy grass pollen (Phleum pratense); (ii) 14 patients who had received 3 or 4 years of conventional immunotherapy followed by 3 years of placebo treatment; (iii) 12 matched seasonal rhinitic patients who had never received immunotherapy; and (iv) 17 non-atopic normal controls. PBMC were stimulated with 20 microg/ml and 200 microg/ml P. pratense extract, or 10 microg/ml of Mycobacterium tuberculosis purified protein derivative (PPD), at 2 x 10(6) cells/ml and 5 x 10(6) cells/ml. IL-5 concentrations in culture supernatants collected after 6 days' culture were measured by ELISA. IL-5 production in response to stimulation with P. pratense extract was highly reproducible and was elevated in both of the immunotherapy treated groups and the untreated rhinitics relative to non-atopic controls (P<0.005 for each group relative to non-atopic controls, under each of the four conditions tested). However, no significant reduction was observed in IL-5 production when immunotherapy treated patients were compared with untreated rhinitic controls. Moreover, abrogation of the cutaneous late-phase responses to allergen following treatment was not associated with reduced IL-5 production by allergen-stimulated peripheral blood T cells. Reduced IL-5 production by peripheral blood T cells may not be necessary for immunotherapy to be effective. Local immunodulation of T cell responses may play a role in this form of treatment. PMID- 9353158 TI - Normal human immunoglobulins for intravenous use (IVIg) delay hyperacute xenograft rejection through F(ab')2-mediated anti-complement activity. AB - Xenotransplantation between discordant species leads to a hyperacute rejection mediated by natural antibodies, both of the IgG and IgM isotypes, activation of complement and endothelial cell activation. The combination of these mechanisms leads to a transplant survival of minutes to a few hours. Polyclonal human immunoglobulins for intravenous use (IVIg) from normal donors have proved effective in a number of antibody-mediated disorders, as well as in inflammatory disorders. We demonstrate that administration of IVIg in a guinea pig to rat model of cardiac xenografting can effectively delay hyperacute rejection. This effect is mediated by the F(ab')2 fragments of IVIg, and is correlated to an anti complementary activity. PMID- 9353159 TI - High intratumoural level of cytokines mediates efficient regression of a rat histiocytoma. AB - We have studied the role of cytokines in the spontaneous regression of AK-5 histiocytoma in syngeneic rats. Animals in which the tumour regresses show high levels of cytokines in the serum compared with animals which succumb to the tumour, and levels of IL-2, IL-4, IL-12, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) are significantly higher in tumour tissue of the former. Thus there is an association between rejection of the tumour and the levels of cytokines present in the tumour mass. Our results also suggest a predominant Th1-type of response in those rats that display early tumour rejection. PMID- 9353160 TI - Calcium efflux and influx in f-met-leu-phe (fMLP)-activated human neutrophils are chronologically distinct events. AB - The kinetics of efflux of calcium mobilized from intracellular stores following activation of human neutrophils with the synthetic chemotactic tripeptide, fMLP (1 microM), as well as that of the subsequent store-operated influx of this cation, has been measured by radiometric procedures using 45Ca. These procedures enabled distinction between net efflux and influx of 45Ca. Preincubation of neutrophils in medium containing 45Ca as the sole source of Ca2+, followed by activation with fMLP, resulted in a rapid efflux of the cation, which coincided with its release from intracellular stores. Efflux terminated at approximately 30 s after addition of fMLP to neutrophils and resulted in the loss of 42 +/- 3% (P < 0.005) of cell-associated 45Ca. Net influx of 45Ca, which was insensitive to the voltage-dependent Ca2+ channel blockading agent, verapamil (20 microM), could only be detected at 30-60 s after the addition of fMLP to neutrophils, and proceeded for about 5 min, resulting in intracellular concentrations of Ca2+ which were 27 +/- 3% (P<0.05) higher than preactivation levels. These results demonstrate that the efflux of cytoplasmic Ca2+ mobilized from intracellular stores during activation of neutrophils by fMLP, and the subsequent influx of extracellular Ca2+ to replete these stores, are chronologically distinct events in fMLP-activated neutrophils. PMID- 9353161 TI - Chemoattractant-induced release of elastase by lipopolysaccharide (LPS)-primed neutrophils; inhibitory effect of the anti-inflammatory drug nimesulide. AB - Human neutrophils, pre-exposed to low concentrations (1-10 ng/ml) of bacterial LPS in the presence of 1% autologous serum, released elastase activity in response to N-formyl-met-leu-phe (fMLP). Both cell incubation with LPS without subsequent fMLP stimulus and fMLP stimulation without prior exposure to LPS failed to promote significant elastase release. Therefore, LPS primes neutrophils for the subsequent release of elastase in response to fMLP. Compared with fMLP, human recombinant C5a had a slight although not significant activity, whereas other chemoattractants such as IL-8, platelet-activating factor and leukotriene B4 were ineffective. The fMLP-induced response of LPS-primed neutrophils was susceptible to suppression by the methane-sulphonanilide anti-inflammatory drug nimesulide and RO 20-1724, which selectively inhibit cAMP-catabolizing phosphodiesterase type IV. This suggests that the elastase release by LPS-primed neutrophils is likely to be controlled by intracellular cAMP, and raises the possibility of limiting pharmacologically the elastase-mediated tissue injury during neutrophilic inflammation. PMID- 9353162 TI - Artificial antigen-presenting cells engineered by recombinant vaccinia viruses expressing antigen, MHC class II, and costimulatory molecules elicit proliferation of CD4+ lymphocytes in vitro. AB - The current study was designed to test the ability of recombinant Vaccinia virus (rVV) encoding essential components of an artificial antigen-presenting cell to activate antigen-specific T cells in vitro. We have constructed a set of rVV encoding separately or in combination a CD4+ T cell-specific epitope (the 133-145 peptide of chicken conalbumin), the MHC class II molecule I-Ak, and costimulatory molecules (mB7-1 and mB7-2). Cultured cells infected with rVV encoding both the antigen and the presenting MHC, but not either one alone, could activate cloned CD4+ T cells specific for the virus-encoded epitope. Additional co-expression of mB7-1 and mB7-2 resulted in further enhancement of T cell response. Thus, our rVV vector expressing four different foreign gene products elicited the highest proliferation rates of antigen-specific cloned T cells. PMID- 9353163 TI - Comparison of aging and hypercholesterolemic effects on the sodium inward currents in cardiac myocytes. AB - To study and to compare the hypercholesterolemic and aging effect on the sodium inward currents (I(Na)) in cardiac myocytes, whole-cell clamp recordings were made in single cardiac myocyte isolated from normo- and diet-induced hypercholesterolemic rabbits of different age groups. The cell capacitance of adult and hyperlipidemic myocytes seemed larger than that of young and normolipidemic ones. However, the sodium current density at a holding potential of -80 mV on adult and hypercholesterolemic ventricular sarcolemma was significantly lower than that on young and normolipidemic one (adult hyperlipidemic: -15.3+/-2.4 pA/pF (n=16), adult control: -28.1+/-3.4 pA/pF (n=13), young hyperlipidemic: -39.5+/-5.4 pA/pF (n=19), young control: -67.3+/ 7.8 pA/pF (n=12)). In aging process, this effect was due to a decrease in channel number, a leftward shift in the inactivation potential and a slowing of the time course of recovery. In hypercholesterolemia, however, the major cause was due to the functional change of sodium currents. In addition to decreasing the sodium current magnitude, hypercholesterolemia lowered the threshold for excitation of cardiac myocytes (-50 mV vs -40 mV). In conclusion, aging process depressed the sodium channel activity in ventricular myocytes. In addition to inducing some similar functional alterations of I(Na) as aging process, long-term hypercholesterolemia could also increase the excitability in cardiac myocytes, which was different from aging process. PMID- 9353164 TI - Progression of glomerulosclerosis, renal hypertrophy, and an increased expression of fibronectin in the renal cortex associated with aging and salt-induced hypertension in Dahl salt-sensitive rats. AB - Aging and hypertension are known to be closely related with the pathogenesis and development of glomerulosclerosis. In this study, we examined the time course changes in the glomerulus associated with salt-induced hypertension using the inbred Dahl salt-sensitive rats. For this purpose, 5-week-old Dahl salt-sensitive rats (n=36) were fed either 4% NaCl diet (n=18) or 0.3% NaCl diet (n=18) up to 17 weeks of age. The high salt diet caused a dramatic increase in systolic blood pressure and also a dramatic renal hypertrophy as shown by a significant increase in the kidney weight. Histological examination revealed an age-dependent progression of glomerulosclerosis as documented by a quantitative scoring. This age-dependent progression was further accelerated by the co-existence of salt induced hypertension in the high salt diet group. Northern blot analysis revealed an increase in the steady state mRNA levels of fibronectin, an important component of mesangial matrices, in the renal cortex, but not in the renal medulla, only in salt-loaded Dahl salt-sensitive rats. These findings indicate that salt-induced hypertension accelerates the age-dependent progression of glomerulosclerosis in Dahl salt-sensitive rats, and fibronectin may play a role in the pathogenesis, development, and progression of glomerulosclerosis associated with salt-induced hypertension. PMID- 9353165 TI - Antinociceptive effects of morphine and U-50,488H on vaginal distension in the anesthetized rat. AB - The antinociceptive activity of the kappa- and mu-opioid receptor agonists, (+/-) U-50,488H and morphine, was examined in a vaginal distension model in anaesthetized female rats. Vaginal distension induced a reproducible cardiovascular response (CVR) which was inhibited in a dose related manner by morphine (0.03-1.0 mg/kg i.v., ED50 = 0.16 mg/kg) and (+/-)-U-50,488H (0.08-1.6 mg/kg i.v., ED50 = 0.49 mg/kg). Morphine (0.3 microg/rat) administered i.c.v. inhibited the CVR by 81.6 +/- 7.9% whereas (+/-)-U-50,488H (30-300 microg/rat) was inactive by this route. A low dose of naloxone (30 microg/kg i.v.) blocked the effect of morphine but not that of (+/-)-U-50,488H. The kappa-opioid antagonist, nor-binaltorphimine (10 mg/kg s.c.) abolished the response to (+/-)-U 50,488H but not that of morphine. This demonstrates that both central and peripheral mu-opioid receptors may be involved in morphine-induced antinociception whereas the kappa-opioid agonist, (+/-)-U-50,488H, blocks vaginal nociception by acting on peripheral kappa-opioid receptors. PMID- 9353166 TI - Apoptosis in haemopoietic progenitor cells exposed to extremely low-frequency magnetic fields. AB - Epidemiological studies have indicated a modestly increased risk for the development of acute myeloid leukaemia in children who live close to high-voltage power-lines. Recent evidence has suggested that a common property shared by a number of known and suspected tumour promoters is their ability to block the process of apoptosis. Therefore, one possible mechanistic explanation for the apparent leukaemogenic effect of weak, low-frequency magnetic fields, such as emitted by power-lines and electrical appliances, would be their expression of tumour-promoting activity by interfering with the regulation of apoptosis in multipotent haemopoietic progenitor cells. In order to test this hypothesis, we have employed the well-characterized multipotential haemopoietic progenitor cell line FDCP-mix(A4). These cells are non-leukaemic and undergo apoptosis when deprived of appropriate growth factors such as Interleukin-3. We have tested a series of different regimes of weak, low-frequency magnetic fields: nulled fields, Ca2+-ion cyclotron resonance conditions at 50 Hz, and vertical 50 Hz fields of 6 microT(RMS), 1 mT(RMS) and 2 mT(RMS), exposing the cells for 2 hours, 24 hours, 4 days or 7 days under various culture conditions. We have not seen any significant alteration in apoptosis induced by any of the exposure regimes tested. We therefore conclude that the regulation of viability and apoptosis in FDCP-mix(A4) cells is not disturbed by weak magnetic fields of the magnitude and type indicated. PMID- 9353167 TI - Bovine fetuin is an inhibitor of insulin receptor tyrosine kinase. AB - Fetuin has been identified earlier as the bovine homolog of the human plasma protein, alpha2-Heremans Schmid glycoprotein (alpha2-HSG). Although bovine fetuin shares over 70% amino acid sequence similarity with alpha2-HSG and rat fetuin, no common function(s) have been identified. We report that immunoaffinity purified bovine fetuin acts as an inhibitor of insulin receptor tyrosine kinase activity (IR-TKA) with half-maximal inhibition at 1.5 microM. In vitro, bovine fetuin (1.5 microM) blocked insulin-induced autophosphorylation of the human IR completely and the half-maximal inhibitory effect was observed at 0.5 microM. Incubation of HIRcB cells (rat1 fibroblasts transfected with wild-type human insulin receptor cDNA) with bovine fetuin (1.5 microM) inhibited insulin-induced tyrosine phosphorylation of the IR beta-subunit by 40%. In addition, bovine fetuin (2 microM) completely blocked insulin-stimulated DNA synthesis in H-35 rat hepatoma cells. Our results, together with earlier reports on rat fetuin and human alpha2 HSG, indicate a common IR-TK inhibitory function for fetuin homologs. PMID- 9353168 TI - Non-exocytotic GABA overflow in rat striatum inhibits gnawing. AB - The present study tested the hypotheses that spontaneous gamma-aminobutyric acid (GABA) efflux in anterior rat striatum is 1) independent of intra- and extracellular calcium; and 2) is physiologically relevant. Extracellular dopamine (DA) and GABA were sampled from striatum of awake, freely moving rats using in vivo microdialysis. Although dialysate concentrations of DA were 2 to 3 times greater than GABA and were decreased by at least 70% by removal of calcium, GABA was unaffected even in the presence of EGTA or the intracellular calcium chelator APTRA-AM. Functional significance of this non-exocytotic pool of GABA was tested by injecting 3-mercaptopropionic acid (3-MPA), an inhibitor of GABA synthesis, into the striatum via a guide cannula sidled alongside a microdialysis probe and measuring subsequent effects on behavior and perfusate concentrations of GABA. Results show that 3-MPA increases gnawing behavior suggesting that basal, non exocytotic GABA overflow normally functions to suppress gnawing. PMID- 9353169 TI - Effects of selective cyclooxygenase-2 inhibitors on alkaline secretory and mucosal ulcerogenic responses in rat duodenum. AB - Effects of the selective cyclooxygenase-2 (COX-2) inhibitors such as NS-398 and nimesulide on duodenal HCO3- secretory and ulcerogenic responses to mucosal acidification were examined in rats, in comparison with indomethacin, a nonselective COX inhibitor. Duodenal HCO3- secretion in anesthetized rats was increased in response to mucosal acidification. The increased HCO3- response to acid was significantly suppressed by pretreatment with indomethacin (10 mg kg( 1), s.c.), while both NS-398 and nimesulide (10 mg kg(-1), s.c.) had no effect on this response. The luminal release of prostaglandin E2 (PGE2) was increased during and after mucosal acidification, and this response was significantly inhibited by indomethacin but not NS-398 or nimesulide. Indomethacin provoked hemorrhagic lesions in the duodenum when acid hypersecretion was concomitantly induced by histamine (8 mg kg(-1) hr(-1), i.v.), while either NS-398 or nimesulide did not cause damage in the duodenum. Either of these drugs had no effect on histamine-induced acid secretion. On the other hand, both NS-398 and nimesulide showed a significant suppression against carrageenan-induced rat paw edema, similar to indomethacin. The present study supports a mediator role for endogenous PGs in duodenal HCO3- secretion in response to mucosal acidification and suggests that COX-1 but not COX-2 is a key enzyme in regulating this process and maintaining the mucosal integrity against acid in the duodenum. PMID- 9353170 TI - Lithium deficiency reduces thymus weight and alters differential blood count in rats. AB - Effects of lithium (Li) deficiency and/or immobilization stress on the thymus weight and differential blood count of rats were studied. The thymus weight of the rats fed a low Li diet were lighter than those of rats fed a Li supplemented diet, whether the rats were exposed to stress or not. Of the rats not exposed to stress, those in the low Li diet group showed a significant decrease in the ratios of neutrophils and T helper lymphocytes. However, there was an increase in the total number of lymphocytes in the low Li dietary group. It was shown that Li deficiency altered the responses to stress in rats. PMID- 9353172 TI - The protective effect of class III antiarrhythmic agents against calcium overload in cultured myocytes. AB - Calcium ions have been implicated in the mechanisms of ventricular arrhythmias. Impairment of intercellular coupling by calcium overload is considered to facilitate ventricular fibrillation (VF) and to sup-press its self termination. According to our hypothesis, any compound that decreases intracellular calcium concentration [Ca2+]i during VF can serve as defibrillating drug. In this study, we examined the effect of d-sotalol and tedisamil on calcium overload in cultured, spontaneously beating rat cardiomyocytes. The changes of [Ca2+]i were measured by indo-1 method and the intercellular synchronization by image analysis. The results showed that increase in [Ca2+]o from 1.9 mM to 3.9 mM increased [Ca2+]i from 100 nM to 320 nM and transformed the synchronized cell movement to an asynchronous one. Administration of 5 x 10(-6) M d-sotalol or 10( 6) M tedisamil, decreased the [Ca2+]i to its basic level and restored the synchronized activity. In summary: Our results showed that increase in [Ca2+]i known to cause inhibition of intercellular coupling, that could lead to arrhythmia and fibrillation while d-sotalol or tedisamil prevented this effect. These results support our hypothesis, that class III antiarrhythmic compounds with positive inotropic effect, increase intercellular synchronization, by decreasing free [Ca2+]i, most probably by increasing the Ca2+ uptake by the sarcoplasmic reticulum, and therefore act as a defibrillating compound. PMID- 9353171 TI - Antinociceptive properties of the hydroalcoholic extract and preliminary study of a xanthone isolated from Polygala cyparissias (Polygalaceae). AB - Polygala cyparissias (Polygalaceae) grows abundantly on Brazil's Atlantic coast, belonging to the typical underbrush vegetation of dunes and have been used in folk medicine for treatment of several diseases, such as disturbances of bowel and kidney. The hydroalcoholic extract of P. cyparissias (HE, 3 to 60 mg kg(-1), i.p. or 25 to 200 mg kg(-1), p.o.) produced significant and graded inhibition of acetic acid-induced abdominal constrictions, with mean ID50 values of 6 and 72 mg kg(-1), respectively. The HE (at this same range of doses) also produced dose related inhibition of both the early and the late phase of formalin-induced licking. The calculated mean ID50 values for the early phase were: >60 and >200 mg kg(-1), while for the late phase they were 11 and 101 mg kg(-1), respectively, by i.p. and p.o. routes. The HE also caused dose-related inhibition of formalin induced edema formation (P<0.01). The HE (3 to 60 mg kg(-1), i.p. or 50 to 200 mg kg(-1), p.o.) produced significant and dose-related inhibition of the neurogenic nociception caused by topical injection of capsaicin, with mean ID50 values of 12 and 71 mg kg(-1), respectively. Given orally, the HE (50 to 200 mg kg(-1)) prevented in a dose-dependent manner, bradykinin (3 nmol/paw) and substance P (10 nmol/paw)-induced hyperalgesia in the rat paw, with mean ED50 values of 122 and 121 mg kg(-1), respectively, but was ineffective in the hot-plate model of nociception. The antinociception caused by the HE, in contrast to that of morphine (5 mg kg(-1), s.c.), was not reversed by naloxone (5 mg kg(-1), i.p.) when assessed in the acetic acid writhing test. The HE, at antinociceptive doses, did not affect motor coordination of animals when assessed in the rota-rod model. The xanthone isolated from P. cyparissias, identified as 1,7-dihydroxy-2,3 dimethoxy xanthone (0.3 to 30 mg kg(-1), i.p.), produced dose-related inhibition of acetic acid-induced abdominal constriction, with mean ID50 value of 1.5 mg kg( 1). These data show that the active principle(s) present in the HE of P. cyparissias, elicited pronounced antinociception when assessed by i.p. or p.o. routes, against both inflammatory and neurogenic nociception, and was able to prevent bradykinin and substance P-induced hyperalgesia. Its precise mechanism of action still remains unclear. PMID- 9353174 TI - Evidence for direct cellular protective effect of PL-10 substances (synthesized parts of body protection compound, BPC) and their specificity to gastric mucosal cells. AB - The direct gastric mucosal cellular effect of four PL-10 substances (a synthesized part of human body protection compound, BPC containing 14 or 15 amino acids) was studied on freshly isolated rat gastric mucosal cells and on a mouse myeloma cell line (Sp2/0-Ag14) in an ethanol-induced cell injury model. The examined substances were not toxic for the cells. Two of them proved to be significantly protective against the direct cellular damaging effect of ethanol (PL 10.1.15AK-3 in 5 microg/ml dose and PL 10.1.AK14-2 dose-dependently, ED50=50 ng/ml) on gastric mucosal cells. This cytoprotective effect was failured on mouse myeloma cells. Based on these results a part of the in vivo protection induced by BPC seems to be a direct cellular protective effect to gastric mucosal cells. PMID- 9353173 TI - Fluoxetine's effects on ethanol's rewarding, aversive and stimulus properties. AB - These experiments examined the influence of fluoxetine on ethanol-induced conditioned place preference, ethanol-induced conditioned taste aversion, and ethanol discrimination. In the place conditioning experiment, male Swiss-Webster mice received 4 pairings of a distinctive floor cue with 2 g/kg ethanol, 10 mg/kg fluoxetine + ethanol, or fluoxetine alone. A different floor was paired with saline. During conditioning ethanol produced locomotor stimulation. Fluoxetine + ethanol resulted in greater levels of locomotor activity during conditioning trials 2-4. Fluoxetine alone also caused increases in activity. Floor preference testing revealed conditioned place preference in groups receiving ethanol. Fluoxetine did not change the magnitude of ethanol-induced conditioned place preference nor produced place conditioning alone. In the taste conditioning procedure, mice received 1-h access to 0.2 M NaCl solution followed by injections of 0, 5 or 10 mg/kg fluoxetine and 0 or 2.5 g/kg ethanol. Ethanol produced reductions in NaCl intake. Fluoxetine (10 mg/kg) enhanced the development of ethanol-conditioned taste aversion but did not cause taste aversion alone. In the ethanol discrimination experiment, mice were trained to respond for 10% sucrose on an FR20 schedule following injections of either 1 g/kg ethanol or saline. Following acquisition, 10 mg/kg fluoxetine pretreatment enhanced ethanol appropriate responding at a dose of ethanol (0.5 g/kg) below the training dose. These results indicate enhancement of serotonergic activity influences ethanol aversion and discrimination but not ethanol reward. PMID- 9353175 TI - Attenuation of mecamylamine-precipitated nicotine-withdrawal aversion by the 5 HT3 receptor antagonist ondansetron. AB - The effect of ondansetron (0.01-0.1 mg/kg, s.c.), a selective 5-HT3 receptor antagonist, on mecamylamine-precipitated nicotine-withdrawal aversion was examined in the conditioned place preference paradigm. Male Sprague-Dawley rats were chronically treated subcutaneously with 9 mg/kg/day (-)-nicotine tartrate using an osmotic minipump. After nicotine treatment for 7 days, mecamylamine (1 mg/kg, s.c.), a nicotinic receptor antagonist, produced place aversion in nicotine-dependent rats. This aversive effect was dose-dependently antagonized by pretreatment with ondansetron 30 min prior to the conditioning. These results suggest that ondansetron may attenuate the place aversion associated with nicotine withdrawal, and may be useful for the treatment of nicotine dependence. PMID- 9353176 TI - Quantitative structure-activity relationships (QSARs) for estrogen binding to the estrogen receptor: predictions across species. AB - The recognition of adverse effects due to environmental endocrine disruptors in humans and wildlife has focused attention on the need for predictive tools to select the most likely estrogenic chemicals from a very large number of chemicals for subsequent screening and/or testing for potential environmental toxicity. A three-dimensional quantitative structure-activity relationship (QSAR) model using comparative molecular field analysis (CoMFA) was constructed based on relative binding affinity (RBA) data from an estrogen receptor (ER) binding assay using calf uterine cytosol. The model demonstrated significant correlation of the calculated steric and electrostatic fields with RBA and yielded predictions that agreed well with experimental values over the entire range of RBA values. Analysis of the CoMFA three-dimensional contour plots revealed a consistent picture of the structural features that are largely responsible for the observed variations in RBA. Importantly, we established a correlation between the predicted RBA values for calf ER and their actual RBA values for human ER. These findings suggest a means to begin to construct a more comprehensive estrogen knowledge base by combining RBA assay data from multiple species in 3D-QSAR based predictive models, which could then be used to screen untested chemicals for their potential to bind to the ER. Another QSAR model was developed based on classical physicochemical descriptors generated using the CODESSA (Comprehensive Descriptors for Structural and Statistical Analysis) program. The predictive ability of the CoMFA model was superior to the corresponding CODESSA model. PMID- 9353177 TI - Human cancer syndromes: clues to the origin and nature of cancer. AB - More than 20 different hereditary cancer syndromes have now been defined and attributed to specific germline mutations in various inherited cancer genes. Collectively, the syndromes affect about 1 percent of cancer patients. An individual who carries a mutant allele of an inherited cancer gene has a variable risk of cancer that is influenced by the particular mutation, other cellular genes, and dietary, lifestyle, and environmental factors. Though hereditary cancer syndromes are rare, their study has provided powerful insights into more common forms of cancer. Somatic mutations in sporadic cancers frequently alter the inherited cancer genes, and the functions of cell signaling pathways have been illuminated by study of the affected genes. Further investigation of inherited mutations that affect susceptibility to cancer will aid efforts to effectively prevent, detect, and treat the disease. PMID- 9353178 TI - Genetic testing for cancer risk. AB - Genetic testing for cancer susceptibility is already part of the clinical management of families with some of the well-defined (but uncommon) inherited cancer syndromes. In cases where the risks associated with a predisposing mutation are less certain, or where there is no clearly effective intervention to offer those with a positive result, its use is more controversial. Careful evaluation of costs and benefits, and of the efficacy of interventions in those found to be at risk, is essential and is only just beginning. An immediate challenge is to ensure that both health professionals and the public understand clearly the issues involved. PMID- 9353179 TI - Nucleic acid-based methods for the detection of cancer. AB - Continued elucidation of the genetic changes that drive cancer progression is yielding new and potentially powerful nucleic acid-based markers of neoplastic disease. Pilot studies indicate that these markers can be used to detect cancer cells in a variety of clinical settings with unprecedented precision. Nucleic acid-based markers may prove to be valuable tools for early detection of cancer in asymptomatic individuals, for confirmation or exclusion of a cancer diagnosis that is based on suspicious but nondiagnostic clinical material, for assessment of tumor burden in cancer patients, and for assessment of response to preventive approaches applied to healthy individuals who are at high risk of developing cancer. Examples of these markers, their potential applications, and the current practical limitations on their clinical use are reviewed here. PMID- 9353180 TI - Oncogenic transcription factors in the human acute leukemias. AB - Chromosomal translocations in the human acute leukemias rearrange the regulatory and coding regions of a variety of transcription factor genes. The resultant protein products can interfere with regulatory cascades that control the growth, differentiation, and survival of normal blood cell precursors. Support for this interpretation comes from the results of gene manipulation studies in mice, as well as the sequence homology of oncogenic transcription factors with proteins known to regulate embryonic development in primitive organisms, including the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster. Many of these genetic alterations have important prognostic implications that can guide the selection of therapy. The insights gained from studies of translocation generated oncogenes and their protein products should hasten the development of highly specific, and hence less toxic, forms of leukemia therapy. PMID- 9353181 TI - Integrating genetic approaches into the discovery of anticancer drugs. AB - The discovery of anticancer drugs is now driven by the numerous molecular alterations identified in tumor cells over the past decade. To exploit these alterations, it is necessary to understand how they define a molecular context that allows increased sensitivity to particular compounds. Traditional genetic approaches together with the new wealth of genomic information for both human and model organisms open up strategies by which drugs can be profiled for their ability to selectively kill cells in a molecular context that matches those found in tumors. Similarly, it may be possible to identify and validate new targets for drugs that would selectively kill tumor cells with a particular molecular context. This article outlines some of the ways that yeast genetics can be used to streamline anticancer drug discovery. PMID- 9353182 TI - Environment and cancer: who are susceptible? AB - Acting in concert with individual susceptibility, environmental factors such as smoking, diet, and pollutants play a role in most human cancer. However, new molecular evidence indicates that specific groups-characterized by predisposing genetic traits or ethnicity, the very young, and women-may have heightened risk from certain exposures. This is illustrated by molecular epidemiologic studies of environmental carcinogens such as polycyclic aromatic hydrocarbons and aromatic amines. Individual genetic screening for rare high-risk traits or for more common, low-penetrant susceptibility genes is problematic and not routinely recommended. However, knowledge of the full spectrum of both genetic and acquired susceptibility in the population will be instrumental in developing health and regulatory policies that increase protection of the more susceptible groups from risks of environmental carcinogens. This will necessitate revision of current risk assessment methodologies to explicitly account for individual variation in susceptibility to environmental carcinogens. PMID- 9353183 TI - Recent advances in chemoprevention of cancer. AB - Chemoprevention is the use of pharmacologic or natural agents that inhibit the development of invasive cancer either by blocking the DNA damage that initiates carcinogenesis or by arresting or reversing the progression of premalignant cells in which such damage has already occurred. Recent advances in our understanding of the mechanisms of carcinogenesis have led to the synthesis of new drugs that can inhibit tumor development in experimental animals by selective action on specific molecular targets, such as the estrogen, androgen, and retinoid receptors or inducible cyclooxygenase. Several of these agents (including tamoxifen, 13-cis-retinoic acid, retinyl palmitate, and an acyclic retinoid) are clinically effective in preventing the development of cancer, particularly in patients who are at high risk for developing second primary tumors after surgical removal of the initial tumor. PMID- 9353184 TI - Mapping the inside of the ribosome with an RNA helical ruler. AB - The structure of ribosomal RNA (rRNA) in the ribosome was probed with hydroxyl radicals generated locally from iron(II) tethered to the 5' ends of anticodon stem-loop analogs (ASLs) of transfer RNA. The ASLs, ranging in length from 4 to 33 base pairs, bound to the ribosome in a messenger RNA-dependent manner and directed cleavage to specific regions of the 16S, 23S, and 5S rRNA chains. The positions and intensities of cleavage depended on whether the ASLs were bound to the ribosomal A or P site, and on the lengths of their stems. These data predict the three-dimensional locations of the rRNA targets relative to the positions of A- and P- site transfer RNAs inside the ribosome. PMID- 9353189 TI - Direct measurement of distances and angles in biomolecules by NMR in a dilute liquid crystalline medium. AB - In isotropic solution, internuclear dipolar couplings average to zero as a result of rotational diffusion. By dissolving macromolecules in a dilute aqueous nematic discotic liquid-crystalline medium containing widely spaced magnetically oriented particles, a tunable degree of solute alignment with the magnetic field can be created while retaining the high resolution and sensitivity of the regular isotropic nuclear magnetic resonance (NMR) spectrum. Dipolar couplings between 1H 1H, 1H-13C, 1H-15N, and 13C-13C pairs in such an oriented macromolecule no longer average to zero, and are readily measured. Distances and angles derived from dipolar couplings in human ubiquitin are in excellent agreement with its crystal structure. The approach promises to improve the accuracy of structures determined by NMR, and extend the size limit. PMID- 9353192 TI - A euryarchaeal lysyl-tRNA synthetase: resemblance to class I synthetases. AB - The sequencing of euryarchaeal genomes has suggested that the essential protein lysyl-transfer RNA (tRNA) synthetase (LysRS) is absent from such organisms. However, a single 62-kilodalton protein with canonical LysRS activity was purified from Methanococcus maripaludis, and the gene that encodes this protein was cloned. The predicted amino acid sequence of M. maripaludis LysRS is similar to open reading frames of unassigned function in both Methanobacterium thermoautotrophicum and Methanococcus jannaschii but is unrelated to canonical LysRS proteins reported in eubacteria, eukaryotes, and the crenarchaeote Sulfolobus solfataricus. The presence of amino acid motifs characteristic of the Rossmann dinucleotide-binding domain identifies M. maripaludis LysRS as a class I aminoacyl-tRNA synthetase, in contrast to the known examples of this enzyme, which are class II synthetases. These data question the concept that the classification of aminoacyl-tRNA synthetases does not vary throughout living systems. PMID- 9353193 TI - Role of sensory-evoked NMDA plateau potentials in the initiation of locomotion. AB - Reticulospinal (RS) neurons constitute the main descending motor system of lampreys. This study reports on natural conditions whereby N-methyl-D-aspartate (NMDA)-mediated plateau potentials were elicited and associated with the onset of locomotion. Reticulospinal neurons responded in a linear fashion to mild skin stimulation. With stronger stimuli, large depolarizing plateaus with spiking activity were elicited and were accompanied by swimming movements. Calcium imaging revealed sustained intracellular calcium rise upon sensory stimulation. Blocking NMDA receptors on RS neurons prevented the plateau potentials as well as the associated rise in intracellular calcium. Thus, the activation of NMDA receptors mediates a switch from sensory-reception mode to a motor command mode in RS neurons. PMID- 9353194 TI - Structural plasticity in a remodeled protein-protein interface. AB - Remodeling of the interface between human growth hormone (hGH) and the extracellular domain of its receptor was studied by deleting a critical tryptophan residue (at position 104) in the receptor, creating a large cavity, and selecting a pentamutant of hGH by phage display that fills the cavity and largely restores binding affinity. A 2.1 A resolution x-ray structure of the mutant complex showed that the receptor cavity was filled by selected hydrophobic mutations of hGH. Large structural rearrangements occurred in the interface at sites that were distant from the mutations. Such plasticity may be a means for protein-protein interfaces to adapt to mutations as they coevolve. PMID- 9353195 TI - A low-barrier hydrogen bond in the catalytic triad of serine proteases? Theory versus experiment. AB - Cleland and Kreevoy recently advanced the idea that a special type of hydrogen bond (H-bond), termed a low-barrier hydrogen bond (LBHB), may account for the "missing" transition state stabilization underlying the catalytic power of many enzymes, and Frey et al. have proposed that the H-bond between aspartic acid 102 and histidine 57 in the catalytic triad of serine proteases is an example of a catalytically important LBHB. Experimental facts are here considered regarding the aspartic acid-histidine and cis-urocanic H-bonds that are inconsistent with fundamental tenets of the LBHB hypothesis. The inconsistencies between theory and experiment in these paradigm systems cast doubt on the existence of LBHBs, as currently defined, within enzyme active sites. PMID- 9353196 TI - Inhibition of brain Gz GAP and other RGS proteins by palmitoylation of G protein alpha subunits. AB - Palmitoylation of the alpha subunit of the guanine nucleotide-binding protein Gz inhibited by more than 90 percent its response to the guanosine triphosphatase (GTPase)-accelerating activity of Gz GAP, a Gz-selective member of the regulators of G-protein signaling (RGS) protein family of GTPase-activating proteins (GAPs). Palmitoylation both decreased the affinity of Gz GAP for the GTP-bound form of Galphaz by at least 90 percent and decreased the maximum rate of GTP hydrolysis. Inhibition was reversed by removal of the palmitoyl group by dithiothreitol. Palmitoylation of Galphaz also inhibited its response to the GAP activity of Galpha-interacting protein (GAIP), another RGS protein, and palmitoylation of Galphai1 inhibited its response to RGS4. The extent of inhibition of Gz GAP, GAIP, RGS4, and RGS10 correlated roughly with their intrinsic GAP activities for the Galpha target used in the assay. Reversible palmitoylation is thus a major determinant of Gz deactivation after its stimulation by receptors, and may be a general mechanism for prolonging or potentiating G-protein signaling. PMID- 9353197 TI - Areal segregation of face-processing neurons in prefrontal cortex. AB - A central issue in cognitive neuroscience concerns the functional architecture of the prefrontal cortex and the degree to which it is organized by sensory domain. To examine this issue, multiple areas of the macaque monkey prefrontal cortex were mapped for selective responses to visual stimuli that are prototypical of the brain's object vision pathway-pictorial representations of faces. Prefrontal neurons not only selectively process information related to the identity of faces but, importantly, such neurons are localized to a remarkably restricted area. These findings suggest that the prefrontal cortex is functionally compartmentalized with respect to the nature of its inputs. PMID- 9353198 TI - Thermoregulation in the mouths of feeding gray whales. AB - Vascular structures for heat conservation in the tongue of the gray whale (Eschrichtius robustus) are reported here. Numerous individual countercurrent heat exchangers are found throughout the massive tongue. These converge at the base of the tongue to form a bilateral pair of retia. Temperature measurements from the oral cavity of a live gray whale indicate that more heat may be lost through the blubber layer over the body than through the tongue, despite the fact that the tongue is far more vascularized and has much less insulation. These heat exchangers substantially reduce heat loss when these whales feed in cold waters. PMID- 9353199 TI - A mRNA signal for the type III secretion of Yop proteins by Yersinia enterocolitica. AB - Pathogenic Yersinia species have a specialized secretion system (type III) to target cytotoxic Yop proteins during infection. The signals of YopE and YopN sufficient for the secretion of translational reporter fusions were mapped to the first 15 codons. No common amino acid or peptide sequence could be identified among the secretion signals. Systematic mutagenesis of the secretion signal yielded mutants defective in Yop translation; however, no point mutants could be identified that specifically abolished secretion. Frameshift mutations that completely altered the peptide sequences of these signals also failed to prevent secretion. Thus, the signal that leads to the type III secretion of Yop proteins appears to be encoded in their messenger RNA rather than the peptide sequence. PMID- 9353201 TI - Isoxaben soil biodegradation in pear tree orchard after repeated high dose application. AB - During the past nine years, each of the plots of a pear tree orchard were treated annually with the same herbicide treatment. The following herbicide treatments were compared, each being made by application of a mixture of two or three herbicides: 1a, no herbicide at all, weeds being hoed (control 1a); 2, diuron + paraquat 3 + 1 kg/ha; 3, simazine + paraquat 2 + 1 kg/ha; 4, isoxaben + diuron + paraquat 0.5 + 1.6 + 1 kg/ha; and 5, isoxaben + simazine + paraquat 0.5 + 1.25 + 1 kg/ha. In March 1996, one year after the final orchard herbicide treatment, isoxaben could not be detected in the soils of any field plots; isoxaben was incorporated at 0.74 mg/kg in the loamy soils sampled separately in each of the field plots, and the soils were incubated in the laboratory. Isoxaben soil half lives were 92 days in the soils treated previously with herbicide treatments 1a, 2, or 3 and 42 days in the soils treated with herbicide treatments 4 and 5. The repeated isoxaben treatments applied in the past thus enhanced the isoxaben soil biodegradation; diuron, simazine, and paraquat had no influence on this rate enhancement. On the other hand, herbicide treatments 4 and 5 were applied in the orchard in April 1996 on the corresponding plots treated in this manner for the last nine years. Isoxaben + paraquat 0.5 + 1 kg/ha was applied simultaneously on other plots (control 1b) not treated in the past with isoxaben. During the growth season in the orchard, the isoxaben soil half-lives in the control plots 1b was 101 days, and 41 days in the plots where herbicide treatments 4 or 5 were applied. PMID- 9353202 TI - Investigations into using the nematode Caenorhabditis elegans for municipal and industrial wastewater toxicity testing. AB - This investigative study assesses the ease and usefulness of the nematode Caenorhabditis elegans for identifying contributors to effluent toxicity within an industrial and municipal wastewater treatment plant (WWTP) system. Several different types of industries, including fiberglass manufacturing, paper packaging, and yarn dyeing, discharge effluent into the municipal wastewater treatment plant, which in turn discharges into a local creek. A major objective of this study was to identify primary sources of toxicity throughout the system with a nematode toxicity test. Twenty-four-hour composite water samples were taken periodically over a ten-month period at five strategic points within the system: (1) at the point of discharge at each of the three industries, (2) at the combined industrial influent of the wastewater treatment plant, (3) at the effluent of the WWTP, (4) upstream of the WWTP discharge, and (5) downstream of the WWTP discharge. Samples were analyzed for basic water chemistry, and each sample was tested for whole effluent toxicity using a 72-h nematode test with mortality as the end point. Results suggest that interactions between the wastewaters of certain industries may increase the overall nematode toxicity in the wastewater treatment facility's composite influent and effluent. Nematode mortality trends indicate relatively high toxicity levels in wastewater entering the WWTP from contributing industries. High WWTP influent toxicity may potentially be due to varying flow rate ratios of industrial discharges, release of varying toxic constituents in wastewaters, and toxic interactions between chemical constituents of industrial wastewaters. The evaluation of toxicity within the treatment system may pinpoint locations where pollution prevention strategies may be implemented to reduce overall toxicity at the point of discharge. PMID- 9353200 TI - PCDD/Fs in soil samples collected in the vicinity of a municipal solid waste incinerator: human health risks. AB - The concentrations of polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) were determined in soil samples taken from 24 sites in the vicinity of a municipal solid waste incinerator (Montcada, Barcelona, Spain). Samples were collected within a radius of 3 km in each of the three main directions of the wind rose in that area. Hepta- and octa-CDDs were the predominant congeners and contributors to TEQ. PCDD/F levels ranged from 0.30 to 44.26 ng TEQ/kg (dry matter), with median and mean values of 3.52 and 6.91 ng TEQ/kg, respectively. The highest and lowest PCDD/F concentrations were found at 750 m (44.26 ng TEQ/kg) and 3000 m (0. 30 ng TEQ/kg) from the stack, while the PCDD/PCDF ratio was 1.78. The health risk analysis of the data shows that the PCDD/F intake from soils is substantially lower than the tolerable daily intake for toxicologic (other than cancer) effects of PCDD/Fs. PMID- 9353205 TI - Long-term recovery of macrobenthos and fish assemblages after water pollution abatement measures in the River Petite Baise (France). AB - Riverine ecosystems are subject to a large variety of man-made influences, and in recent years a new public awareness of the need to protect rivers has emerged within the industrialized world. The present study focuses on the effect of abatement of pollution from one factory on the recovery of fish and macroinvertebrate species richness in the River Petite Baise over a 20-year period (1973-1993). Until 1973, the Petite Baise (75 km in length) received not only factory waste (nitrogenous rich effluents) but also the untreated sewage of many villages and agricultural runoff from the river's catchment. In 1970, macroinvertebrates and fishes were absent in the river despite having initially been classified as of the barbel Barbus barbus zone, and extensive efforts were made by the factory to reduce pollution to allow the riverine community to recover. Regular surveys between 1973 and 1993 revealed a progressive improvement in water quality and the recolonization of the river by macrobenthic and fish populations. Macrobenthic fauna species richness gradually increased from zero in 1970 to level 8 or 9 in 1993 (Verneaux and Tuffery method), reflecting improvements in river water quality. Fish were not observed until 40 km downstream of the factory in 1978, 16 km in 1980, and approximately 5 km by 1990. Fish species richness increased from five in 1978 (barbel, chub Leuciscus cephalus, stone loach Barbatula barbatula, minnow Phoxinus phoxinus, gudgeon Gobio gobio) to eight in 1990 (the carnivorous brown trout Salmo fario, the omnivorous chub, and six benthophagous fishes: gudgeon, barbel, minnow, stone loach, sofie Chondrostoma toxostoma, carp Cyprinus carpio). Fish biomass 45 km downstream the factory increased from 1 g/m2 in 1978 to more than 5 g/m2 by 1990. Recolonization of the river, in particular those areas furthest downstream from the factory, occurred as a result of reductions in the nitrogen inputs emanating from the factory. PMID- 9353207 TI - Effects of nitrite exposure on acid-base balance, respiratory protein, and ion concentrations of giant freshwater prawn Macrobrachium rosenbergii at low pH. AB - Macrobrachium rosenbergii that had been exposed individually for 24 h to 0 (control), 2, 5, 10 mg/L nitrite-N (nitrite as nitrogen) at 4. 3 and 7.7 pH levels were examined for hemolymph nitrite-N, oxyhemocyanin, protein, acid-base balance, ion concentrations, and ammonia-N (ammonia as nitrogen) excretion. Hemolymph oxyhemocyanin, protein, pH, HCO3- , TCO2, osmolality, and ion concentrations were inversely related to ambient nitrite-N concentration and were lower at pH 4.3. However, hemolymph nitrite-N, PO2 and PCO2 levels, and ammonia-N excretion were directly related to ambient nitrite-N, and were higher at pH 4.3. Ambient nitrite-N and pH level interacted to cause changes in hemolymph nitrite N, oxyhemocyanin, protein, PO2, and pH levels. It is concluded that for M. rosenbergii following nitrite exposure, the incorporated nitrite causes a decrease of pH and an increase of PO2 in the hemolymph where it reduces oxyhemocyanin level; disturbs nitrogen excretion, ion regulation, and respiratory gas exchange; and may lead to a decrease of oxygen-carrying capacity, which are affected more at low pH. PMID- 9353208 TI - Concentration-dependent changes of PCB patterns in fish-eating mammals: structural evidence for induction of cytochrome P450. AB - Data sets on CB concentrations in fish-eating mammals from five laboratories were combined to test and refine a pharmacokinetic model. Clear differences in PCB patterns were observed between species. The ability to metabolize chlorobiphenyl (CB) congeners with vicinal H-atoms only in the ortho- and meta-positions and with one ortho-chlorine substituent generally increased in the order otter < cetaceans (harbor porpoise, common dolphin) < phocid seals (harbor and grey seal), but the metabolism of congeners with vicinal H-atoms in the meta- and para positions and with two ortho-chlorines increased in the order cetaceans < seals < otter. Both categories of congeners are probably metabolized by different families of cytochrome P450 (1A and 2B) of which levels apparently differed between the cetaceans, the pinnipeds, and the otter. Within-species CB patterns differed in a concentration-dependent manner. The induction of cytochrome P450 enzymes offers the most likely explanation for this phenomenon, but starvation could have a similar effect on occasion. PMID- 9353209 TI - Mercury accumulation in mink fed fish collected from streams on the Oak Ridge Reservation. AB - This study evaluates effects of feeding mercury (Hg) contaminated fish collected from streams on the Oak Ridge Reservation (ORR) on mink. Diets composed of 25, 50, or 75% fish collected from streams on the ORR were fed to mink beginning 3 months prior to breeding and ending 6 weeks following whelping. Mercury concentrations in diets, tissues of adult mink and their offspring, and physiological and reproductive effects were recorded and compared to concentrations and effects observed in mink fed diets composed of 75% fish collected from the Clinch River above the ORR or from the ocean. Mercury concentrations in prepared diets and in tissues of adult mink and their offspring increased progressively with increased percentage of ORR fish in the diets. Female mink fed diets containing 75% ORR fish had reduced body weight and a decreased number of kits compared to those fed diets containing 75% fish collected above the ORR or from the ocean. However, based on previously reported Hg concentrations associated with adverse effects in mink, the observed adverse effects are not thought to result from exposure to Hg. PMID- 9353210 TI - Determination of polycyclic aromatic hydrocarbons (PAH) and their metabolites in blood, feces, and urine of rats orally exposed to PAH contaminated soils. AB - Polycyclic aromatic hydrocarbons (PAH) have become an ubiquitous upper soil component as a consequence of industrialization involving a multitude of combustion processes. Ingestion of PAH contaminated soil is considered to be a major exposure route, specifically for small children living on these soils. Health risk assessment is based on extrapolations from data obtained via studies performed with pure chemicals. Additionally it is assumed that after oral intake all PAH present in the soil will be absorbed by the human body. Interactions with the soil matrix, however, may modulate the bioavailability of PAH. In this study, we examined the absorption and excretion of PAH in rats orally exposed either to industrially contaminated soils or pure model compounds as anthracene, pyrene and benzo(a)pyrene (B[a]P). The model compounds and the metabolites, 1-hydroxypyrene (1-OH-pyrene) and 3-hydroxybenzo(a)pyrene (3-OH-B[a]P), were measured in blood, feces or urine by means of HPLC with fluorescence detection. Because of rapid biotransformation only minimal levels of unmetabolized anthracene, pyrene and B[a]P in blood could be detected. The pharmacokinetic parameters were nonlinear and suggestive of enterohepatic cycling. Only low levels of the compounds were excreted unchanged in feces whereas the levels of the metabolites were considerably higher in feces and urine. These results indicate that the dosed PAH are largely absorbed by the gastrointestinal tract, subsequently metabolized and excreted as metabolites via urine and feces. Significant differences between the soil-treated group and the pure mixture-treated group could be observed; the soil treated group showed higher fecal excretion of unchanged pyrene (0.5 versus 0.2% of the original dose) and B[a]P (1 versus 0.3%), lower excretion of 1-OH-pyrene in feces (5.1 versus 17. 0%), and lower excretion of 1-OH-pyrene in urine (0.2 versus 3.4%). The fecal excretion of 3-OH-B[a]P between the two groups was similar (8.8 versus 8.8%). These results suggest that the soil matrix is capable of reducing the absorption of at least pyrene. Therefore, exposure risk assessment models assuming complete bioavailability of soilmatrix-bound PAH probably overestimate the endogenous dose. PMID- 9353212 TI - Biological monitoring of the fungicide epoxiconazol during application. AB - A method was developed for the biological monitoring of the fungicide epoxiconazol (Opus; BASF). Comparison of the urine levels of a hydroxylated metabolite after dermal application to the levels after oral intake revealed a dermal absorption of 1-2.5% of the dose. In a field study with 10 applicators a dermal exposure ranging between 60 and 10,000 microgram/person/day was determined from the urine levels of a hydroxylated metabolite; the contribution of the inhalation exposure was found to be negligible. From these data an incorporation of 1 to 100 microgram epoxiconazol/person/day could be derived. The measured exposure was compared to two commonly used exposure models. The model calculation resulted in a dermal exposure of 555 microgram/person/day (German BBA model) and 2115 microgram/person/day (British POEM), respectively, which is in accordance with the actually measured exposure. PMID- 9353211 TI - Applicability of aspecific noninvasive methods for biomonitoring of occupational exposure to deltamethrin: preliminary study using an animal model. AB - Deltamethrin (CAS registry No. 52918-63-5), a synthetic dibromo-pyrethroid insecticide is highly effective against a broad spectrum of insects, and is widely used on crops and in public health programs. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of the present study was to analyze previously demonstrated metabolic changes using aspecific noninvasive methods in rats which are potentially applicable for monitoring occupational exposure. Since human exposure to pesticides occurs not only to active principles but to all chemicals present in a commercial formulation, we tested both the pure compound and a deltamethrin-based commercial formulation. Groups of rats were treated, i.p., consecutively for 7 days. The daily doses tested were 5 and 10 mg/kg body weight for pure deltamethrin, corresponding to volumes of 178.57 and 377.14 microliter/kg body weight for the commercial formulation (containing 2.8% deltamethrin). Urine was analyzed for mutagenic metabolites, thioethers, and D glucaric acid content. Faeces extracts were tested for mutagenicity. Results show that DGA urinary excretion values did not mirror the phase I enzyme induction capability of the insecticide. Results obtained for urinary thioethers do not agree completely with those obtained on the influence of deltamethrin on glutathione S-transferase activity in rat liver. In fact, after administration of the deltamethrin commercial formulation, highest thioether excretion values were obtained during the treatment time for treated animals, as compared to controls. The mean values (+/-SEM) of thioether excretion were 0. 033 +/- 0.002 micromole SH/24 h for control animals, 0.122 +/- 0. 004 and 0.185 +/- 0.025 for the two treatment groups. Thence, thioether determination in urine samples seems to be a suitable aspecific noninvasive method for assessing exposure to deltamethrin based formulations, particularly those containing xylene and mesitylene as solvents, as in the tested formulation. Negative or toxic results obtained in the urinary and faecal mutagenicity test seem to exclude the formation and excretion of mutagenic metabolites following treatment with deltamethrin. PMID- 9353213 TI - Gastric cancer mortality and drinking water qualities in Taiwan. AB - The possible association between the risk of gastric cancer and nitrate and hardness in drinking water from municipal supplies was investigated in a matched case-control study in Taiwan. Data on gastric cancer deaths among eligible residents in Taiwan from 1987 through 1991 (6,766 cases) were obtained from the Bureau of Vital Statistics of the Taiwan Provincial Department of Health. Controls were deaths from other causes (6,766 controls) and were matched individually to the cases by sex, year of birth, and year of death. Data on nitrate-nitrogen (NO3-N) and hardness levels in drinking water throughout Taiwan were collected from the Taiwan Water Supply Corporation (TWSC). The municipality of residence for cases and controls was assumed to be the source of the subject's nitrate and hardness exposure via drinking water. There was no difference in gastric cancer rates between the groups with different levels of nitrate. The odds ratios (95% confidence interval) for death from gastric cancer was 0.95 (0.87-1.03) for the group with water nitrate levels between 0.23 and 0.44 mg/L, and 1.02 (0.93-1.11) for the group with nitrate levels greater than 0.45 mg/L. However, the results show a significant negative relationship between drinking water hardness and gastric cancer mortality. Odds ratios were 1.16 (1.07-1.26) and 1.65 (1.52-1.79), respectively, for exposure to moderately hard water and soft water compared with the use of hard water. This is an important finding for the Taiwan water industry and human health risk. PMID- 9353214 TI - Characteristics and genetic determinants of bacteriocin activities produced by Carnobacterium piscicola CP5 isolated from cheese. AB - Carnobacterium piscicola CP5, isolated from a French mold-ripened soft cheese, produced a bacteriocin activity named carnocin CP5, which inhibited Carnobacterium, Enterococcus and Listeria spp. strains, and among the Lactobacillus spp. only Lactobacillus delbrueckii spp. [24]. The activity was purified by ammonium sulfate precipitation, anion exchange, and hydrophobic interaction chromatography followed by reverse-phase high-performance liquid chromatography (RP-HPLC). This latter step separated two peaks with anti listerial activity (CP51 and CP52). Carnocin CP51 was partially sequenced, and the N-terminal part revealed the presence of the "pediocin-like consensus" sequence-Tyr-Gly-Asn-Gly-Val-. Then, a degenerated 24-mer oligonucleotide probe was constructed from the N-terminal sequence and used to detect the structural gene. It was localized on a plasmid of about 40 kb. Cloning of restriction fragments of this one, followed by DNA sequencing, revealed the presence of the second anti-Listeria bacteriocin gene (CP52). By comparing sequences in data banks and confirming results with PCR reactions, carnocin CP51 shared homologies with carnobacteriocin BM1, and carnocin CP52 was similar to carnobacteriocin B2, both produced by C. piscicola LV17 [2]. However, carnobacteriocin A from C. piscicola LV17 gene was lacking in C. piscicola CP5, and the two microorganisms have been isolated from different ecological environments: C. piscicola CP5 and C. piscicola LV17 were isolated from soft cheese and vacuum-packed meat respectively. This fact could allow different application perspectives for C. piscicola CP5. PMID- 9353215 TI - Induction of Encystment and Poly-beta-Hydroxybutyric Acid Production by Azotobacter chroococcum MAL-201 AB - Azotobacter chroococcum MAL-201, when grown under nitrogen-free conditions with excess glucose, accumulated poly-beta-hydroxybutyric acid amounting to 75% of cell dry weight at the late exponential phase. This led to induction of encystment, which increased steadily with concomitant intracellular degradation of the polymer. Increase in encystment and PHB production were parallel up to 0.5% (wt/vol) glucose. Further increase in glucose reduced cyst formation but enhanced PHB accumulation. Replacement of glucose by n-butyl alcohol and metabolically related compounds identified crotonate as the best encystment inducer followed by beta-hydroxybutyrate and butyrate, but PHB production was inhibited in general. Supplementation of medium with these compounds enhanced the onset of encystment, and only beta-hydroxybutyrate increased PHB accumulation significantly. PMID- 9353216 TI - Multiple bacteriocin production by Leuconostoc mesenteroides TA33a and other Leuconostoc/Weissella strains. AB - Leuconostoc (Lc.) mesenteroides TA33a produced three bacteriocins with different inhibitory activity spectra. Bacteriocins were purified by adsorption/desorption from producer cells and reverse phase high-performance liquid chromatography. Leucocin C-TA33a, a novel bacteriocin with a predicted molecular mass of 4598 Da, inhibited Listeria and other lactic acid bacteria (LAB). Leucocin B-TA33a has a predicted molecular mass of 3466 Da, with activity against Leuconostoc/Weissella (W.) strains, and appears similar to mesenterocin 52B and dextranicin 24, while leucocin A-TA33a, which also inhibited Listeria and other LAB strains, is identical to leucocin A-UAL 187. A survey of other known bacteriocin-producing Leuconostoc/Weissella strains for the presence of the three different bacteriocins revealed that production of leucocin A-, B- and C-type bacteriocins was widespread. Lc. carnosum LA54a, W. paramesenteroides LA7a, and Lc. gelidum UAL 187-22 produced all three bacteriocins, whereas W. paramesenteroides OX and Lc. carnosum TA11a produced only leucocin A- and B-type bacteriocins. PMID- 9353218 TI - Microbial Degradation of Diphenylamine Under Anoxic Conditions AB - Diphenylamine (DPA) was cometabolically degraded in anoxic sediment-water batch enrichments and in cultures of newly isolated sulfate-reducing bacteria. In gas chromatography-mass spectrometry (GC-MS) measurements, aniline was identified as a major breakdown product of the diphenylamine structure. After its identification, aniline was quantified by reversed phase high pressure liquid chromatography (HPLC). The fate of the other carbon ring system remained unclear, because benzene (as a product of reductive cleavage), phenol (as a product of hydrolytic cleavage), and/or other ring cleavage products of diphenylamine were not observed in our experiments with the methods employed. PMID- 9353217 TI - H-NS protein represses transcription of the lux systems of Vibrio fischeri and other luminous bacteria cloned into Escherichia coli. AB - High expression in Escherichia coli of the lux system cistron of a luminous bacteria under its own control has been accomplished only for the Vibrio fischeri lux system at high cell density. Mutation of the hns gene in E. coli has resulted in strong expression of the V. fischeri lux system at low cell density even in an rpoS-deleted strain of E. coli that emits very low levels of luminescence. The E. coli double mutant, MC4110 hns::kan rpoS::tet carrying the lux system of V. fischeri, developed high luminescence from the very early stages of cellular growth, regardless of the presence of deletion mutations in the luxI or luxR genes. Moreover, autoinducer synthesis was restored in the double mutant with the luxR-deleted system. plac-controlled V. fischeri luxCDABE genes missing luxI and luxR were dim in E. coli rpoS mutant cells, but had wild-type levels of light in the hns-deleted strain [MC4110 hns rpoS], showing that expression was independent of lux regulators in the absence of H-NS. DNA gyrase inhibitors and DNA intercalating agents also brought about the restoration of luminescence in the rpoS-deficient strain. High expression of the lux systems of Vibrio harveyi, Photobacterium leiognathi, and Xenorhabdus luminescens in E. coli MC4110 hns rpoS cells compared with that in wild-type or rpoS mutants was also accomplished. Taken together, these data suggest that the H-NS protein inhibits transcription in E. coli of the lux systems of all or most luminous bacteria at the luxC gene as well as in the luxRI region of the V. fischeri lux operon. These DNA regions are highly enriched with homopolymeric stretches of poly d(A) and poly d(T) characterizing curved DNA, a preferable site of H-NS binding. The significance of the new findings in understanding the regulatory control of the bacterial lux system is discussed. PMID- 9353219 TI - Effect of gravity changes on the cyanobacterium Synechocystis sp. PCC 6803. AB - The impact of hypergravity and simulated weightlessness were studied to check whether cyanobacteria perceive changes of gravity as stress. Hypergravity generated by a low-speed centrifuge increased slightly the overall activity of dehydrogenases, but the increase was the same for 90 g and 180 g. The protein pattern did not show qualitative alterations during hypergravity treatment up to 180 g. Cells of Synechocystis PCC 6803 subjected to common stressors like salt, heat, and light clearly accumulated at least four general stress proteins (25, 31, 34, and 63 kDa, respectively). Three of these proteins could also be detected after hypergravity, but in such small amounts that their occurrence could only be taken as a weak indication of stress. Low-molecular-weight stress metabolites were not synthesized in response to hypergravity, indicating that this gravity change was unable to activate the osmotic signal transduction chain. Gravity dependent alterations were observed only during simulated weightlessness (generated by a fast-rotating clinostat). The glutamate/glutamine ratio was significantly shifted toward a higher glutamine portion. Altogether, the results may indicate that moderate changes of gravity were hardly, if ever, sensed as stress by cyanobacteria. PMID- 9353220 TI - Stable Transformation of Chlorella: Rescue of Nitrate Reductase-Deficient Mutants with the Nitrate Reductase Gene AB - Unicellular green algae, like Chlorella, offer a potentially useful system for the expression of heterologous proteins. However, the development of Chlorella as a bioreactor has been delayed owing to the lack of a stable transformation technique. Here we report on the use of micro-projectile bombardment to introduce the nitrate reductase (NR) gene from Chlorella vulgaris into NR-deficient Chlorella sorokiniana mutants, resulting in stable transformants. The stable transformants were able to grow on nitrate medium after repeated passages between selective and nonselective medium and exhibited inducible nitrate reductase activity comparable to that of wild-type cells. Southern analysis suggests homologous recombination occurs with insertion of the wild type gene into the mutated gene and that the genes of the two Chlorellaspecies used are very similar. Specific RNase protection assays, selecting for a poorly conserved region of the gene, identified the presence of the C. vulgaris NR transcript only in the transformed C. sorkiniana mutant and not in the mutant. PMID- 9353221 TI - Genetic variation in two cultures of Bradyrhizobium japonicum 110 differing in their ability to impart drought tolerance to soybean. AB - The polymerase chain reaction with arbitrary primers (RAPD) discriminated between two separately maintained cultures of Bradyrhizobium japonicum USDA 110 differing in symbiotic performance under drought conditions. Since strain 110 is used in inoculum production, the use of RAPD to monitor inoculum cultures could help to preserve their genetic composition and prevent the loss of important symbiotic properties. The use of RAPD could also be extended to other B. japonicum strains currently used in inoculum production. PMID- 9353222 TI - Expression of drug-metabolizing enzymes. AB - A principal advance in the production of drug-metabolizing enzymes has been the development of catalytically self-sufficient cytochrome P450 systems, including additional P450-reductase fusion proteins and Escherichia coli and baculovirus coexpression constructs. Continuing work with glutathione transferases has resulted in the identification of important residues by random mutagenesis screening techniques, as well as in the engineering of model Salmonella typhimurium strains for genotoxicity analysis. PMID- 9353223 TI - Stable insect cell cultures for recombinant protein production. AB - Insect cells are relatively cheap to maintain and are capable of producing accurately translated and correctly processed heterologous proteins. Recent research has focused on the development of improved expression vectors for continuous, high-level production of foreign proteins, including a number of membrane-targeted receptors, in Drosophila and lepidopteran insect cells. Mosquito cells have also been employed for studies on the control of vector-borne diseases, such as malaria. PMID- 9353224 TI - Alphaviruses as expression vectors. AB - Alphavirus vectors have been used for efficient high-level expression of a variety of topologically different proteins, allowing studies of protein transport, localization and functional activity in a broad range of host cells. Complex transmembrane proteins have been produced in large quantities through the establishment of scale-up technology. Alphavirus vectors have also shown promising potential in vaccine production and preliminary gene therapy applications. PMID- 9353225 TI - Eukaryotic protein secretion. AB - The components responsible for protein translocation across the endoplasmic reticulum membrane have been identified and their functions have been clarified in vitro. The structural features of the signal peptide specify the factors and pathways of membrane translocation. Various chaperones and folding enzymes are involved in the folding and quality control of secretory proteins in the lumen. PMID- 9353226 TI - Tag games in yeast: the two-hybrid system and beyond. AB - The yeast Saccharomyces cerevisiae and the one- and two-hybrid systems are essential genetic tools for studying the macromolecular interactions that define all living organisms. Newly developed variations on this theme can now address an even bigger set of questions. Reverse one- and two-hybrid systems can identify factors that dissociate or abrogate defined macromolecular interactions. Different forms of three-hybrid systems can evaluate the complex interplay of proteins with RNAs, peptide ligands, small organic ligands or protein kinases. Finally, the ubiquitin-based split-protein sensor and the Sos recruitment systems promise to overcome some limitations of conventional two-hybrid systems. PMID- 9353227 TI - Adenoviral vectors for gene transfer. AB - Adenoviruses began to be developed into highly effective gene expression vectors in the early 1980s. Recently, the increased interest in utilizing this transfer system in vivo has posed new problems for heterologous gene-transfer, spurring a renewed effort in the field of vector development toward solving the structural, immunological and targeting problems posed by gene therapy applications. PMID- 9353229 TI - Production of recombinant proteins by methylotrophic yeasts. AB - The methylotrophic yeasts Hansenula polymorpha, Pichia pastoris and Candida boidinii have been developed as production systems for recombinant proteins. The favourable and most advantageous characteristics of these species have resulted in an increasing number off biotechnological applications. As a consequence, these species--especially H. polymorpha and P. pastoris--are rapidly becoming the systems of choice for heterologous gene expression in yeast. Recent advances in the development of these yeasts as hosts for the production of heterologous proteins have provided a catalogue of new applications, methods and system components. PMID- 9353228 TI - Baculoviruses as expression vectors. AB - Recent advances in baculovirus expression vector technology include improvements to methods for the selection of recombinant viruses and further developments in virion display vectors. It is now also possible to modify the host cell glycosylation pathway to alter the structure of glycans added to the recombinant polypeptide. Baculovirus vectors also continue to be modified to facilitate gene expression in mammalian cells. PMID- 9353230 TI - Poxviruses as expression vectors. AB - Poxviruses are widely used for the cytoplasmic expression of recombinant genes in mammalian cells. Recent improvements allow high expression and simplify the integration of multiple foreign genes. Vaccinia virus mutants and other poxviruses that undergo abortive infection in mammalian cells are receiving special attention because of their diminished cytopathic effects and increased safety. New replicating and 'non-replicating' vectors, encoding the bacteriophage T7 RNA polymerase for transcription of recombinant genes, have been engineered. PMID- 9353231 TI - Eukaryotic protein processing: endoproteolysis of precursor proteins. AB - Limited endoproteolysis of biologically inactive polypeptide precursors is a general mechanism generating a diversity of biologically active peptides and proteins in all eukaryotic phyla. One of the major recognition motifs involves cleavage at either specific single or pairs of basic residues of the general formula (R/K) - Xn - (R/K) decreases, where n = 0, 2, 4 or 6. Such sites are found in a variety of protein precursors in all eukaryotes, including those of endocrine and neural polypeptide hormones, enzymes, growth factors, receptors, adhesion molecules, viral glycoproteins, coagulation factors and even cell signaling molecules. A family of seven mammalian proteinases responsible for the processing of these proproteins has been recently identified. It comprises the proprotein convertases PC1/PC3, PC2, furin/PACE, PC4, PACE4, PC5/PC6 and PC7/SPC7/LPC/PC8. In a combinatorial fashion, these enzymes determine the cell type and time at which biologically active products are derived from a given inactive precursor protein, thereby profoundly affecting cellular communication, differentiation and metabolic activity. PMID- 9353232 TI - Gene expression systems in the development of high-throughput screens. AB - Recent advances in the development of combinatorial automated chemical synthesis, robotic sample handling, and data collection and analysis have significantly increased the number of compounds available for screening against potential therapeutic targets. The implementation of highly sensitive in vitro biochemical and cell-based high-throughput screening assays is essential to facilitate the rapid identification of selective and potent lead molecules from compound libraries. The ability to easily produce functional proteins in sufficient quantities for in vitro biochemical assays and to devise useful cell-based systems is dependent on the successful application of a variety of gene expression systems. PMID- 9353233 TI - Inducible gene expression in mammalian cells and transgenic mice. AB - Advances in biomedicine have accentuated the need to develop methods to deliberately modulate gene activity. In addition to improved versions of the system based on components of the tetracycline resistance operon, several strategies have recently emerged to control gene function at the transcriptional level. Particularly promising are approaches based on non-mammalian steroid hormones, and on small molecules that bind immunophilins. PMID- 9353234 TI - Plasmid DNA expression systems for the purpose of immunization. AB - DNA vaccines induce immune responses against antigens synthesized in vivo after direct introduction of the DNA's encoding sequences. This unique approach to immunization may overcome deficits of traditional antigen-based approaches and provide safe and effective prophylactic and therapeutic vaccines. DNA vaccines are also useful as a research tool, such as for production of monoclonal antibodies. Efforts are now focusing on understanding the mechanism of antigen presentation and the adjuvant effect of immunostimulatory CpG motifs in the vectors to aid optimization of DNA vaccines. PMID- 9353235 TI - Identification and cloning of differentially expressed genes. AB - Only a few of the methods currently used for identification of differentially expressed genes take advantage of the fact that (near) complete sets of cDNA clones and sequences representing all human and mouse genes will be available for high throughput survey of gene expression. Accordingly, strategies based on hybridization of complex (cDNA or RNA) probes to cDNA microarrays, either on glass slides or on chips, are likely to become increasingly more advantageous. Recognizing, however, that the power of these methods depends upon the availability of such resources, strategies are being pursued to facilitate completion of the ongoing efforts to identify all human and mouse genes. PMID- 9353236 TI - Expression systems for industrial Gram-positive bacteria with low guanine and cytosine content. AB - Recent years have seen an increase in the development of gene expression systems for industrial Gram-positive bacteria with low guanine and cytosine content that belong to the genera Bacillus, Clostridium, Lactococcus, Lactobacillus, Staphylococcus and Streptococcus. In particular, considerable advances have been made in the construction of inducible gene expression systems based on the capacity of these bacteria to utilize specific sugars or to secrete autoinducing peptides that are involved in quorum sensing. These controlled expression systems allow for present and future exploitation of these bacteria as cell factories in medical, agricultural, and food biotechnology. PMID- 9353238 TI - Expression systems: Gene expression systems in the genomics era. PMID- 9353237 TI - Reporter gene expression for monitoring gene transfer. AB - The use of reporters such as green fluorescent protein (GFP) and firefly luciferase permit highly sensitive and nondestructive monitoring of gene transfer and expression. Modifications in GFP which increase intensity and thermostability, as well as alter its spectral qualities, have facilitated the use of GFP in a variety of gene transfer methods. Improvements in imaging technologies and their increased application in biological research have allowed the expanded use of luciferase-based reporters in gene transformation, particularly in genetic screens and in monitoring temporal changes in gene expression. PMID- 9353239 TI - Web alert. Expression systems. PMID- 9353240 TI - Beta-blockers for prophylaxis of bleeding from esophageal varices in cirrhotic portal hypertension. Review of the literature. AB - Hemorrhage from esophageal varices is a life-threatening event in patients with liver cirrhosis. About 40% to 80% of the patients surviving the first bleeding suffer from a recurrence of variceal bleeding within one year. This high recurrence rate substantially contributes to the mortality in patients with liver cirrhosis. Therefore, various treatment regimens both in primary and secondary prophylaxis were studied. Most experience in medical primary prophylaxis was collected with beta-blockers, mainly propranolol. Treating patients with esophageal varices with propranolol significantly reduces the incidence of first variceal bleeding. However, the effect on mortality is marginal, and primary prophylaxis is generally not recommended in these patients. Several studies support the hypothesis, that medical prophylaxis with beta-blockers is more effective in reducing the rate of first esophageal bleeding in patients with a high risk of hemorrhage, such as the presence of very large varices with red spots. A score to assess the individual risk of a given patient to suffer a variceal bleeding would be helpful. As long as such a score is not validated, no general rule for this treatment decision can be given. In secondary prophylaxis, both administration of beta-blockers and endoscopic therapy (sclerotherapy or ligation of the varices) are effective in significantly lowering the rate of re bleeding. However, the effect on mortality was not significant in most studies. Several studies comparing the efficacy of medical prophylaxis and endoscopic treatment showed advantages of the endoscopic therapy with a greater reduction in recurrent bleeding episodes. However, medical prophylaxis with beta-blockers has the important advantage of being immediately effective whereas endoscopic procedures provide the best protection against recurrent bleeding after complete obliteration of the varices. Therefore, in the first weeks and months of endoscopic therapy, the additional treatment with beta-blockers may further reduce the risk of re-bleeding. Only half of all studies on this topic reported a significant advantage of such a combined therapy. Therefore, it seems reasonable to restrict this approach to patients with a high risk of re-bleeding such as patients with large sclerotherapy-derived esophageal ulcers. PMID- 9353241 TI - Urodilatin secretion in salt-loaded Wistar rats. AB - The aim of our study was to investigate whether urodilation (URO, INN: ularitide) is present in rat urine and if URO excretion in the rat is influenced by dietary sodium intake. Therefore, three groups of Wistar rats were placed in metabolic cages where they received different sodium diets for 9 days (0.05%, 0.4%, and 8.0% NaCl, respectively). Food and water intake were determined by weight. At days -4, 2, 5, and 8 blood pressure was measured non invasively using the tail cuff method. After nine days rats were anesthetized and blood was drawn for serum electrolyte, plasma A-type natriuretic peptide (CDD/ANP-99-126), and plasma aldosterone concentration measurements. Using a highly specific antibody against URO combined with high performance liquid chromatography and gel chromatography, we were able to show that a URO-like substance of approx. 3.5 kD that is distinct from CDD/ANP-99-126, brain natriuretic peptide, and C-type natriuretic peptide, is present in rat urine. Sodium chloride loaded rats showed significantly increased urinary excretion rates of URO (p < 0.001), chloride (p < 0.001), sodium (p < 0.001), and the fractional excretion of sodium (p < 0.001). In the plasma, sodium (p < 0.01) and chloride (p < 0.001) increased, while potassium, hematocrit, osmolality, plasma CDD/ANP-99-126, as well as glomerular filtration rate (GFR), and systolic blood pressure did not change. Since CDD/ANP-99-126 is believed to be a natriuretic peptide, it is suggested that CDD/ANP-99-126 might participate in the natriuresis due to high dietary sodium intake. In sodium loaded rats, however, plasma CDD/ANP-99-126 remains unchanged, while URO excretion increases with sodium excretion, independent of GFR and blood pressure. We conclude that URO secretion is stimulated by dietary salt loading and might be involved in the regulation of water and electrolyte metabolism in the rat. PMID- 9353242 TI - Modulation of TNF-alpha secretions by alveolar macrophages and blood monocytes after soot-particle or asbestos fibre exposure. AB - Activated monocytes secrete tumor necrosis factor-alpha (TNF-alpha), whose inflammatory and fibroblast activating characteristics may play a role in the maintenance of pulmonary inflammatory processes and subsequent fibrosis. Human alveolar macrophages (AM) and peripheral blood mononuclear cells (PBMNC) were exposed to soot particles or asbestos fibres in concentrations ranging from 5-50 micrograms/1 x 10(6) cells for 8 hrs in RPMI medium. A culture was established with the exposed monocytes and the remaining cells were used to determine TNF alpha. TNF-alpha was quantified by commercial ELISA-kits. 8 hrs exposure to soot particles and asbestos fibres induced a significant increase in spontaneous TNF alpha release (p < 0.05). Cytotoxicity of monocytes was checked by trypan blue exclusion and lactate dehydrogenase assay, noted values ranging from 0.5%-16.2%. PMID- 9353243 TI - Evaluation of atherosclerosis in patients with central retinal vein occlusion by carotid artery duplex scanning and echocardiography: a prospective case-control study. AB - BACKGROUND AND PURPOSE: Central retinal vein occlusion (CRVO) is a common cause of retinal vascular visual loss second to diabetic retinopathy. Atherosclerotic risk factors are thought to affect vascular flow or cause retinal vascular wall abnormalities, thereby contributing to development of CRVO. Previous studies did not fully evaluate the degree of atherosclerotic disease. The purpose of this study was to determine the degree of atherosclerosis of the carotid artery by duplex scanning and to investigate cardiac manifestations of atherosclerotic risk factors by echocardiography in patients with CRVO. MATERIAL AND METHODS: 39 patients (age 63.1 years [50-84 years], 21 men, 18 women) with CRVO were compared with a control group consisting of 39 individuals (age 59.3 years [49-81 years], 19 men, 20 women) in whom echocardiography was performed to rule out endocarditis. Clinical examination, laboratory testing, carotid artery duplex scanning and echocardiography were performed in all patients. RESULTS: Echocardiography revealed significantly increased prevalence of left ventricular hypertrophy (30.8% in CRVO patients, 5.1% in controls) as a typical sign of hypertensive heart disease in CRVO patients, which is consistent with the increased prevalence of hypertension (HTN) (46.2% in CRVO patients, 15.4% in controls). The prevalence of atherosclerosis of carotid artery and ascending aorta, and all other echocardiographic findings were comparable in CRVO patients and controls: regional wall motion abnormality, left ventricular dilatation, aortic valve calcification, and mitral valve calcification. CONCLUSION: Our study demonstrates that CRVO is not associated with atherosclerosis of large arteries, such as the carotid artery and the ascending aorta. We propose that the retinal artery atherosclerosis seen in most CRVO patients is caused by HTN. PMID- 9353244 TI - Association between different psychotic disorders and the DRD4 polymorphism, but no differences in the main ligand binding region of the DRD4 receptor protein compared to controls. AB - The dopamine D4 receptor gene (DRD4 gene) is one of the important candidate genes for schizophrenia and other psychoses. In humans, several alleles with variable repeat numbers of a 48-base-pair element within the third exon are known. The corresponding receptor proteins differ in their pharmacological properties. It might be possible that specific alleles or genotypes predispose for schizophrenia and other psychoses. The aim of the present study was to investigate and compare the frequency of the DRD4 alleles and genotypes in healthy controls and patients suffering from schizophrenia, schizoaffective or affective disorders. The DRD4 subtypes of 92 controls, 91 patients with schizophrenia, 90 patients with affective and 20 with schizoaffective disorders were identified by a combination of Southern blot technique and PCR. Statistical analysis revealed several significant differences between controls and patients, e.g. an increased frequency of the D4.7 allele among patients with schizophrenia, schizoaffective or unipolar affective disorder. The results indicate a possible role of the DRD4 gene polymorphism in the pathophysiology of psychotic diseases. When a part of the DRD4 gene sequence containing the codons for the most important amino acids for dopamine binding in 9 controls, 9 patients with schizophrenia and 10 with affective disorders were compared, no differences could be found. PMID- 9353245 TI - Intraindividual comparison of three stress tests during the early postinfarction period in stable patients with thrombolysis. AB - The intention of the study was to intraindividually compare the ischemic yield of three stress tests early after acute myocardial infarction. At a large community hospital 107 stable patients who survived acute transmural myocardial infarction after thrombolytic therapy followed by an individual optimized medical treatment, were prospectively investigated by three noninvasive stress tests. All patients received bicycle ergometry, 99mTc perfusion scintigraphy and stress echocardiography within three weeks after the acute event. Each patient underwent diagnostic cardiac catheterization for determination of angiographic data. 99mTc perfusion scintigraphy had the highest rate of positive test results (61%), as compared to bicycle ergometry (32%), stress echocardiography (34%) and stress induced angina in any of the stress tests performed (40%). In 79% of the patients studied, at least one of four ischemic parameters was positive. The combination of bicycle ergometry, stress induced angina and 99mTc perfusion scintigraphy detected myocardial ischemia in 78% of the patients studied. Concordance of at least three positive parameters was seen in only 27%. Intraindividual comparison between positive and negative test results was inconclusive. Only stress echocardiography versus stress-induced angina showed a moderate agreement (kappa = 0.44). Stress-induced angina was the only ischemic parameter which corresponded to the grade of the residual stenosis of the infarct related coronary artery (p < 0.01) and reduced left ventricular function (p < 0.005). These findings show, that concordance of three common stress tests in detecting myocardial ischemia anywhere in patients after acute transmural myocardial infarction and thrombolytic therapy is poor. Stress-echocardiography and stress inducible angina show a moderate agreement. Follow-up studies of these patients are currently performed to clarify prognostic significance and therapeutic consequences of positive test results in these patients. PMID- 9353246 TI - Mechanism and regulation of mRNA polyadenylation. PMID- 9353247 TI - Regulation of the replication initiator protein p65cdc18 by CDK phosphorylation. AB - Cyclin-dependent kinases (CDKs) promote the initiation of DNA replication and prevent reinitiation before mitosis, presumably through phosphorylation of key substrates at origins of replication. In fission yeast, the p65cdc18 protein is required to initiate DNA replication and interacts with the origin recognition complex (ORC) and the p34cdc2 CDK. Here we report that p65cdc18 becomes highly phosphorylated as cells undergo the G1 --> S phase transition. This modification is dependent on p34cdc2 protein kinase activity, as well as six consensus CDK phosphorylation sites within the p65cdc18 polypeptide. Genetic interactions between cdc18+ and the S-phase cyclin cig2+ suggest that CDK-dependent phosphorylation antagonizes cdc18+ function in vivo. Using site-directed mutagenesis, we show that phosphorylation at CDK consensus sites directly targets p65cdc18 for rapid degradation and inhibits its replication activity, as strong expression of a constitutively hypophosphorylated mutant form of p65cdc18 results in large amounts of DNA over-replication in vivo. Furthermore, the over replication phenotype produced by this mutant p65cdc18 is resistant to increased mitotic cyclin/CDK activity, a known inhibitor of over-replication. Therefore, p65cdc18 is the first example of a cellular initiation factor directly regulated in vivo by CDK-dependent phosphorylation and proteolysis. Regulation of p65cdc18 by CDK phosphorylation is likely to contribute to the CDK-driven "replication switch" that restricts initiation at eukaryotic origins to once per cell cycle. PMID- 9353248 TI - Casein kinase II regulation of yeast TFIIIB is mediated by the TATA-binding protein. AB - The highly conserved protein kinase casein kinase II (CKII) is required for efficient Pol III transcription of the tRNA and 5S rRNA genes in Saccharomyces cerevisiae. Using purified factors from wild-type cells to complement transcription extracts from a conditional lethal mutant of CKII we show that TFIIIB is the CKII-responsive component of the Pol III transcription machinery. Dephosphorylation of TFIIIB eliminated its ability to complement CKII-depleted extract, and a single TFIIIB subunit, the TATA-binding protein (TBP), is a preferred substrate of CKII in vitro. Recombinant TBP purified from Escherichia coli is phosphorylated efficiently by CKII and, in the presence of a limiting amount of CKII, is able to substantially rescue transcription in CKII-deficient extract. Our results establish that TBP is a key component of the pathway linking CKII activity and Pol III transcription and suggest that TBP is the target of a CKII-mediated regulatory mechanism that can modulate Pol III transcription. PMID- 9353249 TI - Functionally interacting telomerase RNAs in the yeast telomerase complex. AB - The ribonucleoprotein (RNP) enzyme telomerase from Saccharomyces cerevisiae adds telomeric DNA to chromosomal ends in short increments both in vivo and in vitro. Whether or not telomerase functions as a multimer has not been addressed previously. Here we show, first, that following polymerization, the telomerase RNP remains stably bound to its telomeric oligonucleotide reaction product. We then exploit this finding and a previously reported mutant telomerase RNA to demonstrate that, unexpectedly, the S. cerevisiae telomerase complex contains at least two functionally interacting RNA molecules that both act as templates for DNA polymerization. Here, functional telomerase contains at least two active sites. PMID- 9353250 TI - Normal human chromosomes have long G-rich telomeric overhangs at one end. AB - Telomeres protect the ends of linear chromosomes from degradation and abnormal recombination events, and in vertebrates may be important in cellular senescence and cancer. However, very little is known about the structure of human telomeres. In this report we purify telomeres and analyze their termini. We show that following replication the daughter telomeres have different terminal overhangs in normal diploid telomerase-negative human fibroblasts. Electron microscopy of those telomeres that have long overhangs yields 200 +/- 75 nucleotides of single stranded DNA. This overhang is four times greater than the amount of telomere shortening per division found in these cells. These results are consistent with models of telomere replication in which leading-strand synthesis generates a blunt end while lagging-strand synthesis produces a long G-rich 3' overhang, and suggest that variations in lagging-strand synthesis may regulate the rate of telomere shortening in normal diploid human cells. Our results do not exclude the possibility that nuclease processing events following leading strand synthesis result in short overhangs on one end. PMID- 9353251 TI - CD30-dependent degradation of TRAF2: implications for negative regulation of TRAF signaling and the control of cell survival. AB - CD30 is a cell-surface receptor that can augment lymphocyte activation and survival through its ability to induce the transcription factor NF-kappaB. CD30, however, has also been implicated in the induction of apoptotic cell death of lymphocytes. Here we show that one of the effects of CD30 signal transduction is to render cells sensitive to apoptosis induced by the type 1 tumor necrosis factor receptor (TNFR1). This sensitization is dependent on the TRAF-binding sites within the CD30 cytoplasmic domain. One of the proteins that binds to these sites is TRAF2, a signal transduction molecule that is also utilized by TNFR1 to mediate the activation of several downstream kinases and transcription factors. During CD30 signal transduction, we found that binding of TRAF2 to the cytoplasmic domain of CD30 results in the rapid depletion of TRAF2 and the associated protein TRAF1 by proteolysis. These data suggest a model in which CD30 limits its own ability to transduce cell survival signals through signal-coupled depletion of TRAF2. Depletion of intracellular TRAF2 and its coassociated proteins also increased the sensitivity of the cell to undergoing apoptosis during activation of death-inducing receptors such as TNFR1. Consistent with this hypothesis, expression of a dominant-negative form of TRAF2 was found to potentiate TNFR1-mediated death. These studies provide a potential mechanism through which CD30, as well as other TRAF-binding members of the TNFR superfamily, can negatively regulate cell survival. PMID- 9353252 TI - Cooperative effects of INK4a and ras in melanoma susceptibility in vivo. AB - The familial melanoma gene (INK4a/MTS1/CDKN2) encodes potent tumor suppressor activity. Although mice null for the ink4a homolog develop a cancer-prone condition, a pathogenetic link to melanoma susceptibility has yet to be established. Here we report that mice with melanocyte-specific expression of activated H-rasG12V on an ink4a-deficient background develop spontaneous cutaneous melanomas after a short latency and with high penetrance. Consistent loss of the wild-type ink4a allele was observed in tumors arising in ink4a heterozygous transgenic mice. No homozygous deletion of the neighboring ink4b gene was detected. Moreover, as in human melanomas, the p53 gene remained in a wild-type configuration with no observed mutation or allelic loss. These results show that loss of ink4a and activation of Ras can cooperate to accelerate the development of melanoma and provide the first in vivo experimental evidence for a causal relationship between ink4a deficiency and the pathogenesis of melanoma. In addition, this mouse model affords a system in which to identify and analyze pathways involved in tumor progression against the backdrop of genetic alterations encountered in human melanomas. PMID- 9353253 TI - Rescue of osteoclast function by transgenic expression of kinase-deficient Src in src-/- mutant mice. AB - The Src tyrosine kinase has been implicated in a wide variety of signal transduction pathways, yet despite the nearly ubiquitous expression of c-src, src /- mice show only one major phenotype-osteopetrosis caused by an intrinsic defect in osteoclasts, the cells responsible for resorbing bone. To explore further the role of Src both in osteoclasts and other cell types, we have generated transgenic mice that express the wild-type and mutated versions of the chicken c src proto-oncogene from the promoter of tartrate resistant acid phosphatase (TRAP), a gene that is expressed highly in osteoclasts. We demonstrate here that expression of a wild-type transgene in only a limited number of tissues can fully rescue the src-/- phenotype. Surprisingly, expression of kinase-defective alleles of c-src also reduces osteopetrosis in src-/- animals and partially rescues a defect in cytoskeletal organization observed in src-/- osteoclasts. These results suggest that there are essential kinase-independent functions for Src in vivo. Biochemical examination of osteoclasts from these mice suggest that Src may function in part by recruiting or activating other tyrosine kinases. PMID- 9353254 TI - Yeast heat shock mRNAs are exported through a distinct pathway defined by Rip1p. AB - We reported previously that heat or ethanol shock in Saccharomyces cerevisiae leads to nuclear retention of most poly(A)+ RNA but heat shock mRNAs (encoding Hsp70 proteins Ssa1p and Ssa4p) are efficiently exported in a process that is independent of the small GTPase Ran/Gsp1p, which is essential for most nucleocytoplasmic transport. To gain further insights into proteins essential or nonessential for export of heat shock mRNAs, in situ hybridization analyses to detect mRNA and pulse-labeling of proteins were used to examine several yeast mutant strains for their ability to export heat shock mRNAs following stress. Rip1p is a 42-kD protein associated with nuclear pore complexes and contains nucleoporin-like repeat sequences. It is dispensable for growth of yeast cells under normal conditions, but we report that it is essential for the export of heat shock mRNAs following stress. When SSA4 mRNA was induced from a GAL promoter in the absence of stress, it was efficiently exported in a strain lacking RIP1, indicating that Rip1p is required for export of heat shock mRNAs only following stress. Npl3p, a key mediator of export of poly(A)+ RNA, was not required for heat shock mRNA export, whereas Rss1p/Gle1p, a NES-containing factor essential for poly(A)+ RNA export, was also required for export of heat shock mRNAs after stress. High-level expression of the HIV-1 Rev protein, but not of Rev mutants, led to a partial block in export of heat shock mRNAs following stress. The data suggest a model wherein the requirement for Npl3p defines the mRNA export pathway, the requirement for Rip1p defines a pathway used for export of heat shock mRNAs after stress, and additional factors, including Rss1p/Gle1p and several nucleoporins (Rat7p/Nup159p, Rat2p/Nup120p, and Nup145p/Rat10p), are required in both pathways. PMID- 9353255 TI - The yeast nucleoporin rip1p contributes to multiple export pathways with no essential role for its FG-repeat region. AB - The FG-repeat domain of the yeast Rip1 protein (Rip1p) was identified initially as a possible target for the nuclear export signal (NES) of the HIV-1 Rev protein in a yeast two-hybrid assay. Rip1p is inessential, associated with nuclear pore complexes, and structurally related to the FG-nucleoporin family of pore proteins. It contributes to HIV-1 Rev-mediated RNA export and is also important for the export of heat shock RNAs at 42 degrees C. We show here that Rip1p is essential for the export of heat shock RNAs, and this function is fulfilled by the unique carboxyl terminus of Rip1p with no substantial contribution from the FG-repeat region. Genetic interactions between Rip1p and the RNA export mediator Gle1p are described, which support a role of the carboxyl terminus of Rip1p in poly(A)+ RNA export. Finally, this domain of Rip1p also contributes to Rev mediated RNA export. The data suggest that Rip1p promotes the nuclear export of different classes of substrates by contributing to optimal pore function. PMID- 9353256 TI - c-Myc promotes differentiation of human epidermal stem cells. AB - The epidermis contains two types of proliferative keratinocyte: stem cells, with unlimited self-renewal capacity, and transit amplifying cells, those daughters of stem cells that are destined to withdraw from the cell cycle and terminally differentiate after a few rounds of division. In a search for factors that regulate exit from the stem cell compartment, we constitutively expressed c-Myc in primary human keratinocytes by use of wild-type and steroid-activatable constructs. In contrast to its role in other cell types, activation of c-Myc in keratinocytes caused a progressive reduction in growth rate, without inducing apoptosis, and a marked stimulation of terminal differentiation. Keratinocytes can be enriched for stem or transit amplifying cells on the basis of beta1 integrin expression and by use of this method to fractionate cells prior to c-Myc activation, we found that c-Myc acted selectively on stem cells, driving them into the transit amplifying compartment. As a result, activation of c-Myc in epidermis reconstituted on a dermal equivalent led to premature execution of the differentiation program. The transcriptional regulatory domain of c-Myc was required for these effects because a deletion within that domain acted as a dominant-negative mutation. Our results reveal a novel biological role for c-Myc and provide new insights into the mechanism regulating epidermal stem cell fate. PMID- 9353257 TI - end-1 encodes an apparent GATA factor that specifies the endoderm precursor in Caenorhabditis elegans embryos. AB - The endoderm in the nematode Caenorhabditis elegans is clonally derived from the E founder cell. We identified a single genomic region (the endoderm-determining region, or EDR) that is required for the production of the entire C. elegans endoderm. In embryos lacking the EDR, the E cell gives rise to ectoderm and mesoderm instead of endoderm and appears to adopt the fate of its cousin, the C founder cell. end-1, a gene from the EDR, restores endoderm production in EDR deficiency homozygotes. end-1 transcripts are first detectable specifically in the E cell, consistent with a direct role for end-1 in endoderm development. The END-1 protein is an apparent zinc finger-containing GATA transcription factor. As GATA factors have been implicated in endoderm development in other animals, our findings suggest that endoderm may be specified by molecularly conserved mechanisms in triploblastic animals. We propose that end-1, the first zygotic gene known to be involved in the specification of germ layer and founder cell identity in C. elegans, may link maternal genes that regulate the establishment of the endoderm to downstream genes responsible for endoderm differentiation. PMID- 9353258 TI - Identification of three regions essential for interaction between a sigma-like factor and core RNA polymerase. AB - The cyclic interactions that occur between the subunits of the yeast mitochondrial RNA polymerase can serve as a simple model for the more complex enzymes in prokaryotes and the eukaryotic nucleus. We have used two-hybrid and fusion protein constructs to analyze the requirements for interaction between the single subunit core polymerase (Rpo41p), and the sigma-like promoter specificity factor (Mtf1p). We were unable to define any protein truncations that retained the ability to interact, indicating that multiple regions encompassing the entire length of the proteins are involved in interactions. We found that 9 of 15 nonfunctional (petite) point mutations in Mtf1p isolated in a plasmid shuffle strategy had lost the ability to interact. Some of the noninteracting mutations are temperature-sensitive petite (ts petite); this phenotype correlates with a precipitous drop in mitochondrial transcript abundance when cells are shifted to the nonpermissive temperature. One temperature-sensitive mutant demonstrated a striking pH dependence for core binding in vitro, consistent with the physical properties of the amino acid substitution. The noninteracting mutations fall into three widely spaced clusters of amino acids. Two of the clusters are in regions with amino acid sequence similarity to conserved regions 2 and 3 of sigma factors and related proteins; these regions have been implicated in core binding by both prokaryotic and eukaryotic sigma-like factors. By modeling the location of the mutations using the partial structure of Escherichia coli sigma70, we find that two of the clusters are potentially juxtaposed in the three-dimensional structure. Our results demonstrate that interactions between sigma-like specificity factors and core RNA polymerases require multiple regions from both components of the holoenzymes. PMID- 9353259 TI - A bacterial group II intron encoding reverse transcriptase, maturase, and DNA endonuclease activities: biochemical demonstration of maturase activity and insertion of new genetic information within the intron. AB - The Lactococcus lactis group II intron Ll.ltrB is similar to mobile yeast mtDNA group II introns, which encode reverse transcriptase, RNA maturase, and DNA endonuclease activities for site-specific DNA insertion. Here, we show that the Lactococcal intron can be expressed and spliced efficiently in Escherichia coli. The intron-encoded protein LtrA has reverse transcriptase and RNA maturase activities, with the latter activity shown both in vivo and in vitro, a first for any group II intron-encoded protein. As for the yeast mtDNA introns, the DNA endonuclease activity of the Lactococcal intron is associated with RNP particles containing both the intron-encoded protein and the excised intron RNA. Also, the intron RNA cleaves the sense-strand of the recipient DNA by a reverse splicing reaction, whereas the intron-encoded protein cleaves the antisense strand. The Lactococcal intron endonuclease can be obtained in large quantities by coexpression of the LtrA protein with the intron RNA in E. coli or reconstituted in vitro by incubating the expressed LtrA protein with in vitro-synthesized intron RNA. Furthermore, the specificity of the endonuclease and reverse splicing reactions can be changed predictably by modifying the RNA component. Expression in E. coli facilitates the use of group II introns for the targeting of specific foreign sequences to a desired site in DNA. PMID- 9353260 TI - Rpp1, an essential protein subunit of nuclear RNase P required for processing of precursor tRNA and 35S precursor rRNA in Saccharomyces cerevisiae. AB - The gene for an essential protein subunit of nuclear RNase P from Saccharomyces cerevisiae has been cloned. The gene for this protein, RPP1, was identified by virtue of its homology with a human scleroderma autoimmune antigen, Rpp30, which copurifies with human RNase P. Epitope-tagged Rpp1 can be found in association with both RNase P RNA and a related endoribonuclease, RNase MRP RNA, in immunoprecipitates from crude extracts of cells. Depletion of Rpp1 in vivo leads to the accumulation of precursor tRNAs with unprocessed 5' and 3' termini and reveals rRNA processing defects that have not been described previously for proteins associated with RNase P or RNase MRP. Immunoprecipitated complexes cleave both yeast precursor tRNAs and precursor rRNAs. PMID- 9353261 TI - Chromatin remodeling and the control of gene expression. PMID- 9353262 TI - Promoter escape by RNA polymerase II. Formation of an escape-competent transcriptional intermediate is a prerequisite for exit of polymerase from the promoter. AB - Shortly after initiating promoter-specific transcription in vitro, mammalian RNA polymerase II becomes highly susceptible to arrest in a promoter-proximal region 9-13 base pairs downstream of the transcriptional start site (Dvir, A., Conaway, R. C., and Conaway, J. W. (1996) J. Biol. Chem. 271, 23352-23356). Arrest by polymerase in this region is suppressed by TFIIH in an ATP-dependent reaction (Dvir, A., Conaway, R. C., and Conaway, J. W. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 9006-9010). In this report, we present evidence that, in addition to TFIIH and an ATP cofactor, efficient transcription by RNA polymerase II through this promoter-proximal region requires formation of an "escape-competent" transcriptional intermediate. Formation of this intermediate requires template DNA 40-50 base pairs downstream of the transcriptional start site. This requirement for downstream DNA is transient, since template DNA downstream of +40 is dispensable for assembly of the preinitiation complex, for initiation and synthesis of the first 10-12 phosphodiester bonds of nascent transcripts and for further extension of transcripts longer than approximately 14 nucleotides. Thus, promoter escape requires that the RNA polymerase II transcription complex undergoes a critical structural transition, likely driven by interaction of one or more components of the transcriptional machinery with template DNA 40-50 base pairs downstream of the transcriptional start site. PMID- 9353263 TI - Insulin and epidermal growth factor stimulate a conformational change in Rap1 and dissociation of the CrkII-C3G complex. AB - Insulin and epidermal growth factor (EGF) stimulation of Chinese hamster ovary cells expressing the human insulin and EGF receptors resulted in a time-dependent decrease in the ability of a Rap1 antibody (amino acid epitope 121-136) to immunoprecipitate Rap1 from whole cell detergent extracts. This was due to an apparent masking of Rap1 as heat denaturation of the whole cell detergent extracts (5 min at 100 degrees C) resulted in equal immunoprecipitation of Rap1 with this epitope-specific antibody. The time-dependent change in Rap1 immunoreactivity was paralleled with an insulin-stimulated dissociation of the CrkII-C3G complex. Similarly, EGF treatment also resulted in a time-dependent dissociation of the CrkII-C3G complex that occurred concomitant with the masking of the 121-136 Rap1 epitope. Furthermore, pretreatment of the cells with the tyrosine kinase inhibitor, genistein, decreased both the basal and insulin stimulated tyrosine phosphorylation of CrkII that directly correlated with the amount of CrkII that was immunoprecipitated with C3G. Together, these data suggest that insulin and EGF stimulation result in the dissociation of the CrkII C3G complex, thereby inducing an apparent conformation change in Rap1. PMID- 9353264 TI - Calmodulin activates phosphatidylinositol 3-kinase. AB - Calmodulin and phosphatidylinositol 3-kinase are vital components of a number of common intracellular events. Calmodulin, a ubiquitous Ca2+-dependent effector protein, regulates multiple processes in eukaryotic cells, including cytoskeletal organization, vesicular trafficking, and mitogenesis. Phosphatidylinositol 3 kinase participates in events downstream of the receptors for insulin and other growth factors. Here we demonstrate by coimmunoprecipitation and affinity chromatography that Ca2+/calmodulin associates with Src homology 2 domains in the 85-kDa regulatory subunit of phosphatidylinositol 3-kinase, thereby significantly enhancing phosphatidylinositol 3-kinase activity in vitro and in intact cells. Furthermore, CGS9343B, a calmodulin antagonist, inhibited basal and Ca2+ stimulated phosphorylation of phosphatidylinositol in intact cells. These data demonstrate a novel mechanism for modulating phosphatidylinositol 3-kinase and provide a direct link between components of two fundamental signaling pathways. PMID- 9353265 TI - Interaction of neuronal nitric-oxide synthase with caveolin-3 in skeletal muscle. Identification of a novel caveolin scaffolding/inhibitory domain. AB - Neuronal nitric-oxide synthase (nNOS) has been shown previously to interact with alpha1-syntrophin in the dystrophin complex of skeletal muscle. In the present study, we have examined whether nNOS also interacts with caveolin-3 in skeletal muscle. nNOS and caveolin-3 are coimmunoprecipitated from rat skeletal muscle homogenates by antibodies directed against either of the two proteins. Synthetic peptides corresponding to the membrane-proximal caveolin-3 residues 65-84 and 109 130 and homologous caveolin-1 residues 82-101 and 135-156 potently inhibit the catalytic activity of purified, recombinant nNOS. Purified nNOS also binds to a glutathione S-transferase-caveolin-1 fusion protein in in vitro binding assays. In vitro binding is completely abolished by preincubation of nNOS with either of the two caveolin-3 inhibitory peptides. Interactions between nNOS and caveolin-3, therefore, appear to be direct and to involve two distinct caveolin scaffolding/inhibitory domains. Other caveolin-interacting enzymes, including endothelial nitric-oxide synthase and the c-Src tyrosine kinase, are also potently inhibited by each of the four caveolin peptides. Inhibitory interactions mediated by two different caveolin domains may thus be a general feature of enzyme docking to caveolin proteins in plasmalemmal caveolae. PMID- 9353266 TI - Reactive oxygen intermediates are involved in the induction of CD95 ligand mRNA expression by cytostatic drugs in hepatoma cells. AB - Oxidative stress has been associated with the induction of programmed cell death. The CD95 ligand/receptor system is a specific mediator of apoptosis. We have used the model of drug-induced apoptosis to assess whether the CD95 ligand mRNA is induced by reactive oxygen intermediates. Treatment of HepG2 hepatoma cells with bleomycin induced the production of reactive oxygen intermediates and, as an additional parameter of oxidative stress, resulted in glutathione (GSH) depletion. In parallel, CD95 ligand mRNA expression was induced. In a similar fashion CD95 ligand mRNA expression increased after treatment with H2O2. Additional treatment with the antioxidant and GSH precursor N-acetylcysteine resulted in partial restoration of intracellular GSH levels and in reduced induction of CD95 ligand mRNA. Induction of CD95 ligand mRNA by bleomycin was further reduced by combined treatment with N-acetylcysteine and deferoxamine. These data suggest a direct role of oxygen radicals in the induction of the CD95 ligand. PMID- 9353267 TI - Function of yeast Rad52 protein as a mediator between replication protein A and the Rad51 recombinase. AB - The RAD51 and RAD52 genes of Saccharomyces cerevisiae are key members of the RAD52 epistasis group required for genetic recombination and the repair of DNA double-stranded breaks. The RAD51 encoded product mediates the DNA strand exchange reaction. Efficient strand exchange is contingent upon the addition of the heterotrimeric single-stranded DNA binding factor replication protein A (RPA) after Rad51 has nucleated onto the single-stranded DNA. However, if the single stranded DNA is incubated with Rad51 and RPA simultaneously to mimic what may be expected to occur in vivo, the efficiency of strand exchange decreases dramatically, revealing an inhibitory effect of RPA that is distinct from its stimulatory function. Interestingly, the inclusion of Rad52 protein, which has been purified in this study from yeast cells, restores the efficiency of strand exchange. Thus, Rad52 functions as a co-factor for the Rad51 recombinase, acting specifically to overcome the apparent competition by RPA for binding to single stranded DNA. PMID- 9353268 TI - Preferential interaction of sentrin with a ubiquitin-conjugating enzyme, Ubc9. AB - Sentrin is a ubiquitin-like molecule that has been shown to interact with the death domains of Fas and tumor necrosis factor receptor 1 (TNFR1), PML, Rad51, Rad52, and RanGAP1. We have reported previously that sentrin can be conjugated to other proteins in a manner analogous to protein ubiquitination (Kamitani, T., Nguyen, H. P., and Yeh, E. T. H. (1997) J. Biol. Chem. 272, 14001-14004). Furthermore, the conserved C-terminal Gly-Gly residues are required for sentrinization to occur. To identify enzymes which play a role in sentrinization, the yeast two-hybrid system was used to screen a human placenta cDNA library using sentrin as bait. A strong positive interacting clone was found to contain a cDNA insert encoding the ubiquitin-conjugating enzyme, Ubc9. The interaction between sentrin and Ubc9 required the ubiquitin domain and the C-terminal Gly-Gly residues of sentrin. This interaction appears to be specific because sentrin could only interact weakly with UbcH5B, but could not interact with HHR6B, UbcH6 nor E2-EPF. In vitro translated sentrin could be precipitated by a GST-Ubc9 fusion protein, but not by glutathione S-transferase. A beta-mercaptoethanol sensitive Ubc9-sentrin conjugate could also be identified in the in vitro binding assay. Substitution of the conserved cysteine residue of Ubc9 by serine abolished the formation of the Ubc9-sentrin conjugate. Taken together, Ubc9 is a strong candidate to be the key conjugating enzyme in the sentrinization pathway. PMID- 9353269 TI - Intracellular precursor interleukin (IL)-1alpha, but not mature IL-1alpha, is able to regulate human endothelial cell migration in vitro. AB - The human umbilical vein endothelial cell (HUVEC) has a finite lifespan in vitro, and senescent HUVEC contain elevated levels of the negative growth regulator interleukin (IL)-1alpha. IL-1alpha is translated as a signal peptide sequence less cytosolic 31-kDa precursor (IL-1alpha p), which undergoes proteolytic activation to release the mature carboxyl terminus 17-kDa protein (IL-1alpha m). Both the IL-1alpha p and IL-1alpha m proteins are biologically active as exogenous cytokines. Interestingly, only IL-1alpha p contains a nuclear localization sequence between residues 79 and 85. To further study the role of intracellular IL-1alpha in the regulation of human endothelial cell function, a spontaneous HUVEC transformant was stably transfected with IL-1alpha p, IL-1alpha m, and the IL-1alpha p K82N mutant, which attenuates the nuclear traffic of IL 1alpha p. Interestingly, the IL-1alpha p transfectants were found to have a lower migratory potential than either IL-1alpha m or IL-1alpha p K82N transfectants, and the addition of the IL-1 receptor antagonist did not alter the migration of these cells. Immunofluorescence microscopy demonstrated that only the IL-1alpha p transfectants exhibited prominent staining for beta-catenin-associated cell-to cell contacts, as well as pronounced vimentin intermediate filaments and actin cytoskeleton staining. These data suggest that IL-1alpha p, and not IL-1alpha m, may function as an intracellular regulator of the migratory capacity of the human endothelial cell and that the nuclear localization sequence present within IL 1alpha p may be involved in regulating this function. PMID- 9353270 TI - Chemokine receptor CCR3 function is highly dependent on local pH and ionic strength. AB - The CC chemokine receptor 3 (CCR3) plays an important role in the regulation of the migration of eosinophils, a leukocyte population involved in many inflammatory pathologies including asthma. CCR3 binds to the CC chemokine eotaxin, a promigratory cytokine originally isolated as the key component in a model of eosinophil-induced airway inflammation. We show here that eotaxin/CCR3 binding interactions exhibit a marked sensitivity to relatively small changes in the extracellular environment. In particular, modest variations in the pH and the level of sodium chloride over a range of physiologic and near physiologic conditions had dramatic effects on eotaxin binding and CCR3-mediated cytoplasmic Ca2+ mobilization. These biochemical indices were reflected at the functional level as well; small changes in pH and salt also resulted in striking changes in the migration of primary human eosinophils in vitro. These results reveal that relatively small perturbations in extracellular buffer conditions can yield widely disparate interpretations of CCR3 ligand binding and affinities and suggest that modulation of the tissue microenvironment might be utilized to control the affinity and efficacy of chemokine-mediated cell migration. PMID- 9353271 TI - Coordinate regulation of the expression of the fatty acid transport protein and acyl-CoA synthetase genes by PPARalpha and PPARgamma activators. AB - Intracellular fatty acid (FA) concentrations are in part determined by a regulated import/export system that is controlled by two key proteins, i.e. fatty acid transport protein (FATP) and acyl-CoA synthetase (ACS), which respectively facilitate the transport of FAs across the cell membrane and their esterification to prevent their efflux. The aim of this investigation was to analyze the expression pattern of FATP and ACS and to determine whether their expression was altered by agents that affect FA metabolism through the activation of peroxisome proliferator-activated receptors (PPAR) such as the fibrates and thiazolidinediones. FATP mRNA was ubiquitously expressed, with highest levels being detected in adipose tissue, heart, brain, and testis. Fibrate treatment, which is known to preferentially activate PPARalpha, induced FATP mRNA levels in rat liver and intestine and induced ACS mRNA levels in liver and kidney. The antidiabetic thiazolidinedione BRL 49653, which is a high-affinity ligand for the adipocyte-specific PPARgamma form, caused a small induction of muscle but a robust induction of adipose tissue FATP mRNA levels. BRL 49653 did not affect liver FATP and had a tendency to decrease heart FATP mRNA levels. ACS mRNA levels in general showed a similar pattern after BRL 49653 as FATP except for the muscle where ACS mRNA was induced. This regulation of FATP and ACS expression by PPAR activators was shown to be at the transcriptional level and could also be reproduced in vitro in cell culture systems. In the hepatocyte cell lines AML-12 or Fa 32, fenofibric acid, but not BRL 49653, induced FATP and ACS mRNA levels, whereas in the 3T3-L1 preadipocyte cell line, the PPARgamma ligand induced FATP and ACS mRNA levels quicker than fenofibric acid. Inducibility of ACS and FATP mRNA by PPARalpha or gamma activators correlated with the tissue-specific distribution of the respective PPARs and was furthermore associated with a concomitant increase in FA uptake. Most interestingly, thiazolidinedione antidiabetic agents seem to favor adipocyte-specific FA uptake relative to muscle, perhaps underlying in part the beneficial effects of these agents on insulin-mediated glucose disposal. PMID- 9353272 TI - Regulation of ubiquitin-conjugating enzymes by glutathione following oxidative stress. AB - Upon oxidative stress cells show an increase in the oxidized glutathione (GSSG) to reduced glutathione (GSH) ratio with a concomitant decrease in activity of the ubiquitinylation pathway. Because most of the enzymes involved in the attachment of ubiquitin to substrate proteins contain active site sulfhydryls that might be covalently modified (thiolated) upon enhancement of GSSG levels (glutathiolation), it appeared plausible that glutathiolation might alter ubiquitinylation rates upon cellular oxidative stress. This hypothesis was explored using intact retina and retinal pigment epithelial (RPE) cell models. Exposure of intact bovine retina and RPE cells to H2O2 (0.1-1.7 micromol/mg) resulted in a dose-dependent increase in the GSSG:GSH ratio and coincident dose dependent reductions in the levels of endogenous ubiquitin-activating enzyme (E1) ubiquitin thiol esters and endogenous protein-ubiquitin conjugates and in the ability to form de novo retinal protein-125I-labeled ubiquitin conjugates. Oxidant-induced decrements in ubiquitin conjugates were associated with 60-80% reductions in E1 and ubiquitin-conjugating enzyme (E2) activities as measured by formation of ubiquitin thiol esters. When GSH levels in RPE cells recovered to preoxidation levels following H2O2 removal, endogenous E1 activity and protein ubiquitin conjugates were restored. Evidence that S thiolation of E1 and E2 enzymes is the biochemical link between cellular redox state and E1/E2 activities includes: (i) 5-fold increases in levels of immunoprecipitable, dithiothreitol labile 35S-E1 adducts in metabolically labeled, H2O2-treated, RPE cells; (ii) diminished formation of E1- and E2-125I-labeled ubiquitin thiol esters, oligomerization of E225K, and coincident reductions in protein-125I-labeled ubiquitin conjugates in supernatants from nonstressed retinas upon addition of levels of GSSG equivalent to levels measured in oxidatively stressed retinas; and (iii) partial restoration of E1 and E2 activities and levels of protein-125I labeled ubiquitin conjugates in supernatants from H2O2-treated retinas when GSSG:GSH ratios were restored to preoxidation levels by the addition of physiological levels of GSH. These data suggest that the cellular redox status modulates protein ubiquitinylation via reversible S thiolation of E1 and E2 enzymes, presumably by glutathione. PMID- 9353273 TI - Human corneal keratan sulfates. AB - The keratan sulfate-containing proteoglycans were isolated from fourteen pooled human corneas (thirteen from 61- to 86-year-olds, plus one from a 12-year-old). These proteoglycans were subjected to digestion with the enzyme keratanase II, and the released oligosaccharides, which included nonreducing termini and repeat region oligosaccharides but not linkage regions, were reduced with alkaline borohydride and identified on two separate ion-exchange columns. Both of the latter had been calibrated with samples, most of which had been derived from bovine corneal keratan sulfate (Tai, G.-H., Huckerby, T. N., and Nieduszynski, I. A. (1996) J. Biol. Chem. 271, 23535-23546) and all of which had been fully characterized by NMR spectroscopic analysis. The capping structures identified in human corneal keratan sulfates occurred in the relative proportions: NeuAcalpha(2 6)- >NeuAcalpha(2-3)- >GalNAc(S)beta(1-3)-. The other groups of capping structures which had been identified in bovine corneal keratan sulfate, i.e. NeuGcalpha(2-3)-, NeuGcalpha(2-6)-, GlcNAc(S)beta(1-3)- were absent, although the possibility of the presence of some Galalpha(1-3)- structures could not be excluded. In addition, the human sample showed significantly higher levels of alpha(1-3)-fucosylated repeat region structures than did the bovine sample, and it is not clear whether this reflects a species or age dependence as the bovine corneas were from young animals, whereas the human corneas were predominantly from an older group. The charge densities and keratan sulfate chain sizes of the human and bovine keratan sulfate-containing proteoglycans were seen to be similar. PMID- 9353274 TI - Interactions among inactivating and noninactivating Kvbeta subunits, and Kvalpha1.2, produce potassium currents with intermediate inactivation. AB - Experiments were carried out to determine whether coinjection of Kvalpha1.2 with inactivating and noninactivating Kvbeta subunits would produce currents with intermediate kinetics and channel complexes containing a mixture of these subunits. Upon coexpression with a saturating amount of Kvbeta1.2 and increasing levels of a noninactivating deletion mutant of Kvbeta1.2, we show that macroscopic Kvalpha1.2 currents have levels of fractional inactivation and inactivation time constants that are intermediate between those obtained with either the inactivating Kvbeta1.2 or the noninactivating Kvbeta1.2 mutant. We also find that coexpression of Kvalpha1.2 with saturating amounts of Kvbeta1.2 and the deletion mutant produces a population of single channels with properties intermediate to either the inactivating or noninactivating parental phenotype. Our data can best be explained by the presence of an intermediate population of heterooligomeric channels consisting of Kvalpha1.2 with different combinations of both types of subunits. Since Kvalpha1.2 subunits coexist in cells with inactivating and noninactivating Kvbeta subunits, our findings suggest that heterooligomeric assembly of these subunits occurs to increase the range of K+ current kinetics and expression levels. PMID- 9353275 TI - Transcriptional regulation of the human alpha1a-adrenergic receptor gene. Characterization Of the 5'-regulatory and promoter region. AB - We recently cloned cDNAs encoding three subtypes of human alpha1-adrenergic receptors (alpha1ARs), alpha1a, alpha1b, and alpha1d (Schwinn, D. A., Johnston, G. L., Page, S. O., Mosley, M. J., Wilson, K. H., Worman, N. P., Campbell, S., Fidock, M. D., Furness, L. M., Parry-Smith, D. J., Peter, B., and Bailey, D. S. (1995) J. Pharmacol. Exp. Ther. 272, 134-142) and demonstrated predominance of alpha1aARs in many human tissues (Price, D. T., Lefkowitz, R. J., Caron, M. G., Berkowitz, D., and Schwinn, D. A. (1994) Mol. Pharmacol. 45, 171-175). Several lines of evidence indicate that alpha1aARs are important in clinical diseases such as myocardial hypertrophy and benign prostatic hyperplasia. Therefore, we initiated studies to understand mechanisms underlying regulation of alpha1aAR gene transcription. A genomic clone containing 6.2 kb of 5'-untranslated region of the human alpha1aAR gene was recently isolated. Ribonuclease protection and primer extension assays indicate that alpha1aAR gene transcription occurs at multiple initiation sites with the major site located 696 base pairs upstream of the ATG, where a classic initiator sequence is located. Transfection of luciferase reporter constructs containing varying amounts of 5'-untranslated region into human SK-N-MC neuroblastoma cells indicate that a region extending 125 base pairs upstream from the main transcription initiation site contains full alpha1aAR promoter activity. Furthermore, distinct activator and suppressor elements lie 2-3 and 3-5 kilobase pairs upstream, respectively. Although the alpha1aAR promoter contains neither TATA or CAAT elements, gel shift mobility assays targeting three GC boxes immediately upstream of the main transcription initiation site confirm binding of Sp1. Activity of the alpha1aAR promoter is cell-specific, demonstrating highest activity in cells endogenously expressing alpha1aARs. The human alpha1aAR gene also contains several cis regulatory elements, including several insulin and cAMP response elements. Consistent with these observations, we provide the first evidence that treatment of SK-N-MC cells with insulin and cAMP elevating agents leads to an increase in alpha1aAR expression. In conclusion, these data represent the first characterization of the alpha1aAR gene; our findings should facilitate further studies designed to understand mechanisms regulating alpha1AR subtype-specific expression in healthy and diseased human tissue. PMID- 9353276 TI - Identification of HsORC4, a member of the human origin of replication recognition complex. AB - A new member of human origin recognition complex (ORC) has been cloned and identified as the human homologue of Saccharomyces cerevisiae ORC4. HsORC4 is a 45-kDa protein encoded by a 2.2-kilobase mRNA whose amino acid sequence is 29% identical to ScORC4. HsORC4 has a putative nucleotide triphosphate binding motif that is not seen in ScORC4. HsORC4P also reveals an unsuspected homology to the ORC1-Cdc18 family of proteins. HsORC4 mRNA expression and protein levels remain constant through the cell cycle. HsORC4P is coimmunoprecipitated from cell extracts with another subunit of human ORC, HsORC2P, consistent with it being a part of the putative human origin recognition complex. PMID- 9353277 TI - Phosphorylation of elongation factor 1 and ribosomal protein S6 by multipotential S6 kinase and insulin stimulation of translational elongation. AB - Stimulation of protein synthesis in response to insulin is concomitant with increased phosphorylation of initiation factors 4B and 4G and ribosomal protein S6 (Morley, S. J., and Traugh, J. A. (1993) Biochimie 75, 985-989) and is due at least in part to multipotential S6 kinase. When elongation factor 1 (EF-1) from rabbit reticulocytes was examined as substrate for multipotential S6 kinase, up to 1 mol/mol of phosphate was incorporated into the alpha, beta, and delta subunits. Phosphorylation of EF-1 resulted in a 2-2. 6-fold stimulation of EF-1 activity, as measured by poly(U)-directed polyphenylalanine synthesis. The rate of elongation was also stimulated by approximately 2-fold with 80 S ribosomes phosphorylated on S6 by multipotential S6 kinase. When the rates of elongation in extracts from serum-fed 3T3-L1 cells and cells serum-deprived for 1.5 h were compared, a 40% decrease was observed upon serum deprivation. The addition of insulin to serum-deprived cells for 15 min stimulated elongation to a rate equivalent to that of serum-fed cells. Similar results were obtained with partially purified EF-1, with both EF-1 and ribosomes contributing to stimulation of elongation. These data are consistent with a ribosomal transit time of 3.2 min for serum-deprived cells and 1.6 min following the addition of insulin for 15 min. Taken together, the data suggest that insulin stimulation involves coordinate regulation of EF-1 and ribosomes through phosphorylation by multipotential S6 kinase. PMID- 9353278 TI - The gamma-carboxylation recognition site is sufficient to direct vitamin K dependent carboxylation on an adjacent glutamate-rich region of thrombin in a propeptide-thrombin chimera. AB - The propeptides of the vitamin K-dependent proteins contain a gamma-carboxylation recognition site that is required for gamma-glutamyl carboxylation. To determine whether the propeptide is sufficient to direct carboxylation, two mutant prothrombin species were expressed and characterized with regard to posttranslational gamma-carboxylation. A double point mutant, in which serine substituted for cysteines 17 and 22 disrupted a conserved loop formed by a disulfide bond, was fully carboxylated when expressed in Chinese hamster ovary cells. A propeptide/thrombin chimeric protein, constructed by deleting the Gla, aromatic amino acid stack, and kringle domains of prothrombin, has the signal peptide and propeptide juxtaposed to a glutamate-rich COOH-terminal region of prothrombin, residues 249-530. Of the 8 glutamic acid residues contained within the first 40 residues of the NH2 terminus adjacent to the propeptide, at least seven were fully carboxylated as demonstrated by direct gamma-carboxyglutamic acid analysis of the alkaline hydrolysate and by NH2-terminal sequence analysis. These results indicate that the gamma-carboxylation recognition site within the prothrombin propeptide in a prothrombin propeptide-thrombin chimeric protein is sufficient to direct gamma-carboxylase-catalyzed carboxylation of adjacent glutamic acid residues in a glutamate-rich region of thrombin that is not normally gamma-carboxylated. Furthermore, the disulfide loop in the Gla domain of prothrombin is not required for complete carboxylation. PMID- 9353279 TI - Prostaglandin D synthase in human megakaryoblastic cells. AB - The cytosol fraction of human platelets did not convert prostaglandin (PG) H2 to PGD2. However, a homogenate of human megakaryoblastic CMK cells (precursor cells of platelets) produced PGD2 from PGH2. The PGD synthase activity was localized in the cytosol of CMK cells, and absolutely required glutathione. The catalytic properties and Western and Northern blottings indicated that the enzyme was PGD synthase of the hematopoietic type rather than the lipocalin type. When CMK cells were differentiated to megakaryocytes with phorbol ester along with induction of cyclooxygenase-1, the PGD synthase activity increased about 2-fold for 2 days and then decreased. In another human megakaryoblastic cell line, Dami, the PGD synthase increased about 10-fold by the addition of phorbol ester. Thus, the PGD synthase, which was undetectable in platelets, appeared during differentiation of megakaryoblasts to megakaryocytes. PMID- 9353280 TI - Molecular heterogeneity of phospholipase D (PLD). Cloning of PLDgamma and regulation of plant PLDgamma, -beta, and -alpha by polyphosphoinositides and calcium. AB - Phospholipase D (PLD) has emerged as an important enzyme involved in signal transduction, vesicle trafficking, and membrane metabolism. This report describes the cloning and expression of a new Arabidopsis PLD cDNA, designated PLDgamma, and the regulation of PLDgamma, -beta, and -alpha by phosphatidylinositol 4,5 bisphosphate (PIP2) and Ca2+. The PLDgamma cDNA is 3.3 kilobases in length and codes for an 855-amino acid protein of 95,462 Da with a pI of 6.9. PLDgamma shares a 66% amino acid sequence identity with PLDbeta, but only a 41% identity with PLDalpha. A potential N-terminal myristoylation site is found in PLDgamma, but not in PLDalpha and -beta. Catalytically active PLDgamma was expressed in Escherichia coli, and its activity requires polyphosphoinositides. Both PLDgamma and -beta are most active at microM Ca2+ concentrations, whereas the optimal PLDalpha activity requires mM Ca2+ concentrations. Binding studies showed that the PLDs bound PIP2 in the order of PLDbeta > PLDgamma > PLDalpha. This binding ability correlates with the degree of conservation of a basic PIP2-binding motif located near the putative catalytic site. The binding of [3H]PIP2 was saturable and could be competitively decreased by addition of unlabeled PIP2. Neomycin inhibited the activities of PLDgamma and -beta, but not PLDalpha. These results demonstrate that PLD is encoded by a heterogeneous gene family and that direct polyphosphoinositide binding is required for the activities of PLDgamma and beta, but not PLDalpha. The different structural and biochemical properties suggest that PLDalpha, -beta, and -gamma are regulated differently and may mediate unique cellular functions. PMID- 9353281 TI - Measurement of Ca2+ fluxes during elicitation of the oxidative burst in aequorin transformed tobacco cells. AB - We have employed suspension cultured aequorin-transformed tobacco cells to examine the involvement of Ca2+ in signal transduction of the oxidative burst. Use of cultured cells for this purpose was validated by demonstrating that the cells responded to cold shock quantitatively and qualitatively similarly to the intact transgenic plants from which they were derived. Stimulation of the oxidative burst in the cell suspension was achieved by administration of oligogalacturonic acid, Mas-7 (a peptide known to activate G proteins and Ca2+ fluxes), hypo-osmotic stress, or harpin (a protein from the pathogenic bacterium Erwinia amylovora). The latter failed to promote any detectable increase in cytoplasmic Ca2+ concentration, whereas each of the former three triggered a rapid rise in cytosolic Ca2+ followed by a return within seconds to basal Ca2+ levels. Peak Ca2+ concentrations induced by the former three elicitors were approximately 0.7, 1.4, and 1.3 microM, respectively. Three lines of evidence suggest that the observed Ca2+ pulses are essential to transduction of the oxidative burst signals by their respective elicitors: (i) inhibition of the Ca2+ transients with Ca2+ chelators or Ca2+ channel blockers prevented expression of the oxidative burst, (ii) introduction of exogenous Ca2+ into the same cells initiated the burst even in the absence of other inducers of the response, and (iii) the observed Ca2+ transients often returned to near basal levels well before any H2O2 synthesis could be detected, suggesting that the Ca2+ influx is required to communicate the burst signal but not maintain the defense response. These data suggest that Ca2+ pulses serve frequently, but not invariably, to transduce an oxidative burst signal. PMID- 9353282 TI - Fah1p, a Saccharomyces cerevisiae cytochrome b5 fusion protein, and its Arabidopsis thaliana homolog that lacks the cytochrome b5 domain both function in the alpha-hydroxylation of sphingolipid-associated very long chain fatty acids. AB - A search of the Saccharomyces cerevisiae genome data base for cytochrome b5-like sequences identified a 1.152-kilobase pair open reading frame, located on chromosome XIII at locus YMR272C (FAH1). That gene encodes a putative 384-amino acid protein with an amino-terminal cytochrome b5 domain. The b5 core domain shows a 52% identity and 70% similarity to that of the yeast microsomal cytochrome b5 and a 35% identity and 54% similarity to the b5 core domain of OLE1, the S. cerevisiae Delta-9 fatty acid desaturase. Expression of the S. cerevisiae FAH1 cytochrome b5 domain in Escherichia coli produces a soluble protein that exhibits the typical oxidized versus reduced differential absorbance spectra of cytochrome b5. Sequence analysis of Fah1p reveals other similarities to Ole1p. Both proteins are predicted to have two hydrophobic domains, each capable of spanning the membrane twice, and both have the HX(2-3)(XH)H motifs that are characteristic of membrane-bound fatty acid desaturases. These similarities to Ole1p suggested that Fah1p played a role in the biosynthesis or modification of fatty acids. Disruption of the FAH1 gene in S. cerevisiae did not give any visible phenotype, and there was no observable difference in content or distribution of the most abundant long chain saturated and unsaturated 14-18 carbon fatty acid species. Northern blot analysis, however, showed that this gene is expressed at much lower levels ( approximately 150-fold) than the OLE1 gene, suggesting that it might act on a smaller subset of fatty acids. Analysis of sphingolipid-derived very long chain fatty acids revealed an approximately 40 fold reduction of alpha-HO 26:0 and a complementary increase in 26:0 in the gene disrupted fah1Delta strain. GAL1 expression of the S. cerevisiae FAH1 genes in the fah1Delta strain restores alpha-HO 26:0 fatty acids to wild type levels. Also identified are a number of homologs to this gene in other species. Expression of an Arabidopsis thaliana FAH1 gene, which does not contain the cytochrome b5 domain, in the fah1Delta strain produced an approximately 25-fold increase in alpha-HO 26:0 and reduced the levels of its 26-carbon precursor, suggesting that it functions in very long chain fatty acid hydroxylation using an alternate electron transfer mechanism. PMID- 9353283 TI - The C-terminal subdomain makes an important contribution to the DNA binding activity of the Pax-3 paired domain. AB - The recognition of DNA targets by Pax-3 is achieved through the coordinate use of two distinct helix-turn-helix-based DNA-binding modules: a paired domain, composed of two structurally independent subdomains joined by a short linker, and a paired-type homeodomain. In mouse, the activity of the Pax-3 paired domain is modulated by an alternative splicing event in the paired domain linker region that generates isoforms (Q+ and Q-) with distinct C-terminal subdomain-mediated DNA-binding properties. In this study, we have used derivatives of a classical high affinity paired domain binding site (CD19-2/A) to derive an improved consensus recognition sequence for the Pax-3 C-terminal subdomain. This new consensus differs at six out of eight positions from the C-terminal subdomain recognition motif present in the parent CD19-2/A sequence, and includes a 5'-TT 3' dinucleotide at base pairs 15 and 16 that promotes high affinity binding by both Pax-3 isoforms. However, with a less favorable guanine at position 15, only the Q- isoform retains high affinity binding to this sequence, suggesting that this alternative splicing event might serve to stabilize binding to suboptimal recognition sequences. Finally, mutagenic analysis of the linker demonstrates that both the sequence and the spacing in this region contribute to the enhanced DNA-binding properties of the Pax-3/Q- isoform. Altogether, our studies establish a clear role for the Pax-3 C-terminal subdomain in DNA recognition and, thus, provide insights into an important mechanism by which Pax proteins achieve distinct target specificities. PMID- 9353284 TI - An N-linked glycosylation motif from the noncleaving luteinizing hormone receptor substituted for the homologous region (Gly367 to Glu369) of the thyrotropin receptor prevents cleavage at its second, downstream site. AB - The thyrotropin receptor (TSHR) exists in two forms (single polypeptide and two subunits), whereas the lutropin/chorionic gonadotropin receptor (LH/CGR) is a single chain. Recent data suggest that the TSHR cleaves at two sites. We mutagenized selected chimeric TSH-LH/CGR to localize the cleavage sites in the TSHR. All 23 receptors mutated in the estimated vicinity of the upstream site cleaved into two subunits as determined by 125I-TSH cross-linking to intact cells. In contrast, in a series of mutations homologous to the noncleaving LH/CGR, the downstream TSHR cleavage site localized to three amino acids (GQE367 369). Remarkably, group substitution of these residues, but not substitution of individual residues, abolished cleavage. Moreover, the mutation that prevented cleavage (GQE367-369NET) transposed a motif (NET291-293) that is glycosylated in the LH/CGR. TSHR cleavage or noncleavage after substitution of GQE367-369 with other triplets (AAA, NQE, and NQT) was consistent with a role for N-linked glycosylation at this site. In summary, our data (i) support the concept that the TSHR cleaves at two sites, (ii) relate TSHR residues GQE367-369 to cleavage at the second, downstream site, and (iii) suggest that cleavage or noncleavage at site two is related to N-linked glycosylation. These findings provide new insight into the evolutionary divergence of two closely related receptors. PMID- 9353285 TI - Desensitization of thyrotropin-releasing hormone receptor-mediated responses involves multiple steps. AB - Desensitization and recovery of the inositol 1,4,5-trisphosphate (IP3) and intracellular free calcium concentration ([Ca2+]i) responses to thyrotropin releasing hormone (TRH) were measured in HEK293 cells stably expressing the G protein-coupled TRH receptor. TRH caused a large, rapid, and transient increase in IP3 and a biphasic increase in [Ca2+]i. Desensitization of the TRH response was measured by exposing cells to TRH, washing, and then incubating the cells in hormone-free medium before reintroducing TRH and measuring IP3, [Ca2+]i, and intracellular Ca2+ pool size. When cells were incubated with 1 microM TRH for 10 s or 10 min and reexposed to TRH, there was almost no IP3 or [Ca2+]i increase. The IP3 response recovered first, followed by the [Ca2+]i response. The ionomycin releasable intracellular Ca2+ pool was almost completely depleted by TRH, and pool refilling was slow. Thrombin, endothelin, and carbachol, when combined, stimulated large increases in IP3 and [Ca2+]i, but did not block the IP3 or [Ca2+]i responses to TRH measured 10 min later. In contrast, cells exposed to TRH first responded to combined agonists with a nearly normal increase in IP3, but no rise in [Ca2+]i. Thus, the IP3 response to TRH displays homologous desensitization, whereas the [Ca2+]i response displays heterologous desensitization because depletion of intracellular Ca2+ pools prevents responses to other hormones. PMID- 9353286 TI - Uncoupling of calcium mobilization and entry pathways in endothelin-stimulated pituitary lactotrophs. AB - In cells expressing Ca2+-mobilizing receptors, InsP3-induced Ca2+ release from intracellular stores is commonly associated with extracellular Ca2+ influx. Operation of these two Ca2+ signaling pathways mediates thyrotropin-releasing hormone (TRH) and angiotensin II (AII)-induced prolactin secretion from rat pituitary lactotrophs. After an initial hyperpolarization induced by Ca2+ mobilization from the endoplasmic reticulum (ER), these agonists generated an increase in the steady-state firing of action potentials, further facilitating extracellular Ca2+ influx and prolactin release. Like TRH and AII, endothelin-1 (ET-1) also induced a rapid release of Ca2+ from the ER and a concomitant spike prolactin secretion during the first 3-5 min of stimulation. However, unlike TRH and AII actions, Ca2+ mobilization was not coupled to Ca2+ influx during sustained ET-1 stimulation, as ET-1 induced a long-lasting abolition of action potential firing. This lead to a depletion of the ER Ca2+ pool, a prolonged decrease in [Ca2+]i, and sustained inhibition of prolactin release. ET-1-induced inhibition and TRH/AII-induced stimulation of Ca2+ influx and hormone secretion were reduced in the presence of the L-type Ca2+ channel blocker, nifedipine. Basal [Ca2+]i and prolactin release were also reduced in the presence of nifedipine. Furthermore, TRH-induced Ca2+ influx and secretion were abolished by ET-1, as TRH was unable to reactivate Ca2+ influx and prolactin release in ET-1 stimulated cells. Depolarization of the cells during sustained inhibitory action of ET-1, however, increased [Ca2+]i and prolactin release. These results indicate that L-type Ca2+ channel represents a common Ca2+ influx pathway that controls basal [Ca2+]i and secretion and is regulated by TRH/AII and ET-1 in an opposite manner. Thus, the receptor-mediated uncoupling of Ca2+ entry from Ca2+ mobilization provides an effective control mechanism in terminating the stimulatory action of ET-1. Moreover, it makes electrically active lactotrophs quiescent and unresponsive to other calcium-mobilizing agonists. PMID- 9353287 TI - Generation of anti-apoptotic presenilin-2 polypeptides by alternative transcription, proteolysis, and caspase-3 cleavage. AB - PS2, the chromosome 1 familial Alzheimer's disease gene, has been shown to be involved in programmed cell death by three complementary experimental approaches. Reduction of PS2 protein levels by antisense RNA protects from apoptosis, whereas overexpression of an Alzheimer's PS2 mutant increases cell death induced by several stimuli. In addition, ALG-3, a truncated PS2 cDNA, encodes an artificial COOH-terminal PS2 segment that dominantly inhibits apoptosis. Here we describe a physiological COOH-terminal PS2 polypeptide (PS2s, Met298-Ile448) generated by both an alternative PS2 transcript and proteolytic cleavage. We find that PS2s protects transfected cells from Fas- and tumor necrosis factor alpha (TNFalpha) induced apoptosis. Furthermore, a similar anti-apoptotic COOH-terminal PS2 polypeptide (PS2Ccas) is generated by caspase-3 cleavage at Asp329. These results suggest that caspase-3 not only activates pro-apoptotic substrates but also generates a negative feedback signal in which PS2Ccas antagonizes the progression of cell death. Thus, whereas PS2 is required for apoptosis, PS2s and PS2Ccas oppose this process, and the balance between PS2 and these COOH-terminal fragments may dictate the cell fate. PMID- 9353288 TI - The third transmembrane domain of the serotonin transporter contains residues associated with substrate and cocaine binding. AB - Twenty residues in the third transmembrane domain of the serotonin transporter (SERT) were mutated, one at a time, to cysteine. Almost all of these mutants were fully active for serotonin (5-HT) transport and insensitive to inactivation by the positively charged cysteine reagent [2 (trimethylammonium)ethyl]methanethiosul-fonate (MTSET). Two active mutants, I172C and I179C, were sensitive to rapid inactivation by MTSET but were relatively insensitive to the negatively charged reagent (2 sulfonatoethyl)methanethiosulfonate (MTSES). Inactivation of I172C was blocked by 5-HT and cocaine, but I179C was not similarly protected. Replacement of Tyr-175 with cysteine resulted in a mutant with low transport activity, and, at the neighboring Tyr-176, cysteine replacement completely blocked transport. The Y175C and Y176C mutants were expressed on the cell surface at levels 84% and 69%, respectively, that of wild type (C109A) SERT. Mutants Y175C and Y176C had lower cocaine affinity than C109A, as measured by displacement of the high affinity cocaine analog 2beta-carbomethoxy-3beta-(4-[125I]iodophenyl)tropane (beta-CIT). For Y176C, 5-HT affinity also was decreased. MTSET inactivated beta-CIT binding to I172C and Y176C, but only slightly inhibited binding to I179C and C109A. The MTSET sensitivity of cysteine replacements at positions 172, 176, and 179 was not observed when these positions were replaced with alanine, serine, or methionine. The results suggest that Ile-172, Tyr-176 and Ile-179 are on one face of an alpha helical transmembrane element, and that Ile-172 and Tyr-176 are in proximity to the binding site for 5-HT and cocaine. PMID- 9353289 TI - Phosphorylation by neuronal cdc2-like protein kinase promotes dimerization of Tau protein in vitro. AB - In Alzheimer's disease, the microtubule-associated protein tau forms paired helical filaments (PHFs) that are the major structural component of neurofibrillary tangles. Although tau isolated from PHFs (PHF-tau) is abnormally phosphorylated, the role of this abnormal phosphorylation in PHF assembly is not known. Previously, neuronal cdc2-like protein kinase (NCLK) was shown to phosphorylate tau on sites that are abnormally phosphorylated in PHF-tau (Paudel, H. K., Lew, J., Ali, Z., and Wang, J. H. (1993) J. Biol. Chem. 268, 23512-23518). In this study, phosphorylation by NCLK was found to promote dimerization of recombinant human tau (R-tau) and brain tau (B-tau) purified from brain extract. Chemical cross-linking by disuccinimidyl suberate (DSS), a homobifunctional chemical cross-linker that specifically cross-linked R-tau dimers, and a Superose 12 gel filtration chromatography revealed that R-tau preparations contain mixtures of monomeric and dimeric R-tau species. When the structure of NCLK phosphorylated R-tau was studied by a similar approach, DSS preferentially cross linked the phosphorylated R-tau over the nonphosphorylated R-tau, and the phosphorylated R-tau eluted as a dimeric species from the gel filtration column. Phosphorylated R-tau became resistant to DSS upon dephosphorylation and was recovered as a monomeric species from the gel filtration column. In the presence of a low concentration of dithiothreitol (1.65 microM), R-tau formed disulfide cross-linked R-tau dimers. When compared, phosphorylated R-tau formed more disulfide cross-linked dimers than the nonphosphorylated R-tau. B-tau also was specifically cross-linked to dimers by DSS. When B-tau and NCLK-phosphorylated B tau were treated with DSS, phosphorylated B-tau was preferentially cross-linked over nonphosphorylated counterpart. Taken together, these results suggest that phosphorylation by NCLK promotes dimerization and formation of disulfide cross linked tau dimers, which is suggested to be the key step leading to PHF assembly (Schweers, O., Mandelkow, E.-M., Biernat, J., and Mandelkow, E. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 8463-8467). PMID- 9353290 TI - The oxidative half-reaction of xanthine dehydrogenase with NAD; reaction kinetics and steady-state mechanism. AB - The reaction between reduced xanthine dehydrogenase (XDH) from bovine milk and NAD has been studied in detail. An understanding of this reaction is necessary for a complete description of XDH turnover with its presumed natural electron acceptor and to address the preference of XDH for NAD over oxygen as a substrate. The reaction between pre-reduced XDH and NAD was studied by stopped-flow spectrophotometry. The reaction was found to involve two rounds of oxidation with 2 eq of NAD. The first round goes to completion, and the second round reaches a slightly disfavored equilibrium. Rapid binding of NAD with an apparent Kd of 25 +/- 2 microM is followed by NAD reduction at a rate constant of 130 +/- 13 s-1. NADH dissociation at a rate constant of 42 +/- 12 s-1 completes a round of oxidation. These steps have been successfully tested and modeled to repeat themselves in the second round of oxidation. The association rate constant for NAD binding was estimated to be much greater than any rate constant measured in the oxidation by molecular oxygen, thus explaining how NAD competes with oxygen for reducing equivalents. Rate constants for NAD reduction and NADH dissociation are respectively 21- and 7-fold greater than kcat, indicating that the reductive half-reaction of the enzyme by xanthine is mostly rate-limiting in xanthine/NAD turnover. A steady-state mechanism for XDH is discussed. PMID- 9353291 TI - Processing of mammalian and plant S-adenosylmethionine decarboxylase proenzymes. AB - S-Adenosylmethionine decarboxylase (AdoMetDC) is a pyruvoyl enzyme, and the pyruvate is formed in an intramolecular reaction that cleaves a proenzyme precursor and converts a serine residue into pyruvate. The wild type potato AdoMetDC proenzyme processed much faster than the human proenzyme and did not require putrescine for an optimal rate of processing despite the presence of three acidic residues (equivalent to Glu11, Glu178, and Glu256) that were demonstrated in previous studies to be required for the putrescine activation of human AdoMetDC proenzyme processing (Stanley, B. A., Shantz, L. M., and Pegg, A. E. (1994) J. Biol. Chem. 269, 7901-7907). A fourth residue that is also needed for the putrescine stimulation of human AdoMetDC proenzyme processing was identified in the present studies, and this residue (Asp174) is not present in the potato sequence. The site of potato AdoMetDC proenzyme processing was found to be Ser73 in the conserved sequence, YVLSESS, which is the equivalent of Ser68 in the human sequence. Replacement of the serine precursor with threonine or cysteine by site-directed mutagenesis in either the potato or the human AdoMetDC proenzyme did not prevent processing but caused a significant reduction in the rate. Although the COOH-terminal regions of the known eukaryotic AdoMetDCs are not conserved, only relatively small truncations of 8 residues from the human protein and 25 residues from the potato proenzyme were compatible with processing. The maximally truncated proteins show no similarity in COOH-terminal amino acid sequence but each contained 46 amino acid residues after the last conserved sequence, suggesting that the length of this section of the protein is essential for maintaining the proenzyme conformation needed for autocatalytic processing. PMID- 9353292 TI - Pancreatic beta-cell-specific repression of insulin gene transcription by CCAAT/enhancer-binding protein beta. Inhibitory interactions with basic helix loop-helix transcription factor E47. AB - Chronic exposure of beta-cells to supraphysiologic glucose concentrations results in decreased insulin gene transcription. Here we identify the basic leucine zipper transcription factor, CCAAT/enhancer-binding protein beta (C/EBPbeta), as a repressor of insulin gene transcription in conditions of supraphysiological glucose levels. C/EBPbeta is expressed in primary rat islets. Moreover, after exposure to high glucose concentrations the beta-cell lines HIT-T15 and INS-1 express increased levels of C/EBPbeta. The rat insulin I gene promoter contains a consensus binding motif for C/EBPbeta (CEB box) that binds C/EBPbeta. In non-beta cells C/EBPbeta stimulates the activity of the rat insulin I gene promoter through the CEB box. Paradoxically, in beta-cells C/EBPbeta inhibits transcription, directed by the promoter of the rat insulin I gene by direct protein-protein interaction with a heptad leucine repeat sequence within activation domain 2 of the basic helix-loop-helix transcription factor E47. This interaction leads to the inhibition of both dimerization and DNA binding of E47 to the E-elements of the insulin promoter, thereby reducing functionally the transactivation potential of E47 on insulin gene transcription. We suggest that the induction of C/EBPbeta in pancreatic beta-cells by chronically elevated glucose levels may contribute to the impaired insulin secretion in severe type II diabetes mellitus. PMID- 9353293 TI - Store-operated Ca2+ influx and stimulation of exocytosis in HL-60 granulocytes. AB - This study addresses the role of store-operated Ca2+ influx in the regulation of exocytosis in inflammatory cells. In HL-60 granulocytes, which do not possess voltage-operated Ca2+ channels, the chemotactic peptide fMet-Leu-Phe (fMLP) was able to stimulate store-operated Ca2+ influx and to trigger exocytosis of primary granules. An efficient triggering of exocytosis by fMLP required the presence of extracellular Ca2+ and was inhibited by blockers of store-operated Ca2+ influx. However, receptor-independent activation of store-operated Ca2+ influx through thapsigargin did not trigger exocytosis. fMLP was unable to stimulate exocytosis in the absence of cytosolic free Ca2+ concentration [Ca2+]c elevations. However, a second signal generated by fMLP synergized with store-operated Ca2+ influx to trigger exocytosis and led to a left shift of the exocytosis/[Ca2+]c relationship in ionomycin-stimulated cells. The synergistic fMLP-generated signaling cascade was long-lasting, involved a pertussis toxin-sensitive G protein and a phosphatidylinositol 3-kinase. In summary, store-operated Ca2+ influx is crucial for the efficient triggering of exocytosis in HL-60 granulocytes, but, as opposed to Ca2+ influx through voltage-operated Ca2+ channels in neurons, it is not a sufficient stimulus by itself and requires synergistic receptor-generated signals. PMID- 9353294 TI - A protein phosphatase-1-binding motif identified by the panning of a random peptide display library. AB - An unusually large number of regulatory or targeting proteins that bind to the catalytic subunit of protein phosphatase-1 have been recently reported. This can be explained by their possession of a common protein motif that interacts with a binding site on protein phosphatase-1. The existence of such a motif was established by the panning of a random peptide library in which peptide sequences are displayed on the Escherichia coli bacterial flagellin protein for bacteria that bound to protein phosphatase-1. There were 79 isolates containing 46 unique sequences with the conserved motif VXF or VXW, where X was most frequently His or Arg. In addition, this sequence was commonly preceded by 2-5 basic residues and followed by 1 acidic residue. This study demonstrates that binding to protein phosphatase-1 can be conferred to a protein by the presentation of a peptide motif on a surface loop. This binding motif is found in a number of protein phosphatase-1-binding proteins. PMID- 9353295 TI - Blockade of p38 mitogen-activated protein kinase pathway inhibits inducible nitric-oxide synthase expression in mouse astrocytes. AB - Treatment of mouse astrocyte cultures with combined interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha induced expression of inducible nitric-oxide synthase (iNOS), resulting in sustained release of large amounts of nitric oxide, whereas TNF-alpha and IL-1alpha individually were unable to induce iNOS expression in astrocytes. The role of MAPK cascades and of NF-kappaB activation in the early intracellular signal transduction involved in iNOS transcription in TNF-alpha/IL-1alpha-stimulated astrocytes was investigated. TNF-alpha and IL 1alpha activated all p42/44(MAPK), p38(MAPK), and p54(JNK) pathways as determined by immunoprecipitation kinase assays using specific antibodies and substrates. The p38(MAPK) pathway is specifically involved in TNF-alpha/IL-1alpha-induced iNOS expression, since iNOS protein and nitric oxide release in the presence of a specific inhibitor of p38(MAPK), 4-(4-fluorophenyl)-2-2-(4-hydroxyphenyl)-5-(4 pyridyl)-imidazole (FHPI), were dramatically diminished. In contrast, PD98059, a specific inhibitor of MEK1 had no effect on iNOS expression. p38(MAPK) did not couple NF-kappaB to iNOS transcription, but NF-kappaB had a clear role in iNOS transcription regulation. Northern blot analysis showed that the p38(MAPK) pathway controlled iNOS expression at the transcriptional level, since iNOS mRNA was reduced in the presence of FHPI in TNF-alpha/IL-1alpha-stimulated astrocytes. iNOS expression was investigated with TNF receptor (TNFR)-1- and TNFR-2-deficient mice. The TNF-alpha activity in TNF-alpha/IL-1alpha-stimulated astrocytes was exclusively mediated through TNFR-1, most likely because TNFR-2-mediated signals in astrocytes did not connect to the p38(MAPK) pathway. These data suggest that TNF-alpha/IL-1alpha-induced iNOS expression depends on a yet undetermined second pathway in addition to p38(MAPK). PMID- 9353296 TI - The Aspergillus nidulans cnxABC locus is a single gene encoding two catalytic domains required for synthesis of precursor Z, an intermediate in molybdenum cofactor biosynthesis. AB - The Aspergillus nidulans complex locus, cnxABC, has been shown to be required for the synthesis of precursor Z, an intermediate in the molybdopterin cofactor pathway. The locus was isolated by chromosome walking a physical distance of 65 kilobase pairs from the brlA gene and defines a single transcript that encodes, most likely, a difunctional protein with two catalytic domains, CNXA and CNXC. Mutations (cnxA) affecting the CNXA domain, mutants (cnxC) in the CNXC domain, and frameshift (cnxB) mutants disrupting both domains have greatly reduced levels of precursor Z compared with the wild type. The CNXA domain is similar at the amino acid level to the Escherichia coli moaA gene product, while CNXC is similar to the E. coli moaC product, with both E. coli products encoded by different cistrons. In the wild type, precursor Z levels are 3-4 times higher in nitrate grown cells than in those grown on ammonium, and there is an approximately parallel increase in the 2.4-kilobase pair transcript following growth on nitrate, suggesting nitrate induction of this early section of the pathway. Analysis of the deduced amino acid sequence of several mutants has identified residues critical for the function of the protein. In the CNXA section of the protein, insertion of three amino acid residues into a domain thought to bind an iron-sulfur cofactor leads to a null phenotype as judged by complete loss of activity of the molybdoenzyme, nitrate reductase. More specifically, a mutant has been characterized in which tyrosine replaces cysteine 345, one of several cysteine residues probably involved in binding the cofactor. This supports the proposition that these residues play an essential catalytic role. An insertion of seven amino acids between residues valine 139 and serine 140, leads to a temperature-sensitive phenotype, suggesting a conformational change affecting the catalytic activity of the CNXA region only. A single base pair deletion leading to an in frame stop codon in the CNXC region, which causes a null phenotype, effectively deletes the last 20 amino acid residues of the protein, indicating that these residues are necessary for catalytic function. PMID- 9353297 TI - Cell envelope signaling in Escherichia coli. Ligand binding to the ferrichrome iron receptor fhua promotes interaction with the energy-transducing protein TonB. AB - The ferrichrome-iron receptor of Escherichia coli is FhuA, an outer membrane protein that is dependent upon the energy-coupling protein TonB to enable active transport of specific hydroxamate siderophores, infection by certain phages, and cell killing by the protein antibiotics colicin M and microcin 25. In vivo cross linking studies were performed to establish at the biochemical level the interaction between FhuA and TonB. In an E. coli strain in which both proteins were expressed from the chromosome, a high molecular mass complex was detected when the ferrichrome homologue ferricrocin was added immediately prior to addition of cross-linker. The complex included both proteins; it was absent from strains of E. coli that were devoid of either FhuA or TonB, and it was detected with anti-FhuA and anti-TonB monoclonal antibodies. These results indicate that, in vivo, the binding of ferricrocin to FhuA enhances complex formation between the receptor and TonB. An in vitro system was established with which to examine the FhuA-TonB interaction. Incubation of TonB with histidine-tagged FhuA followed by addition of Ni2+-nitrilotriacetate-agarose led to the specific recovery of both TonB and FhuA. Addition of ferricrocin or colicin M to FhuA in this system greatly increased the coupling between FhuA and TonB. Conversely, a monoclonal antibody that binds near the N terminus of FhuA reduced the retention of TonB by histidine-tagged FhuA. These studies demonstrate the significance of ligand binding at the external surface of the cell to mediate signal transduction across the outer membrane. PMID- 9353298 TI - Penaeidins, a new family of antimicrobial peptides isolated from the shrimp Penaeus vannamei (Decapoda). AB - We report here the isolation of three members of a new family of antimicrobial peptides from the hemolymph of shrimps Penaeus vannamei in which immune response has not been experimentally induced. The three molecules display antimicrobial activity against fungi and bacteria with a predominant activity against Gram positive bacteria. The complete sequences of these peptides were determined by a combination of enzymatic cleavages, Edman degradation, mass spectrometry, and cDNA cloning using a hemocyte cDNA library. The mature molecules (50 and 62 residues) are characterized by an NH2-terminal domain rich in proline residues and a COOH-terminal domain containing three intramolecular disulfide bridges. One of these molecules is post-translationally modified by a pyroglutamic acid at the first position. Comparison of the data obtained from the cDNA clones and mass spectrometry showed that two of these peptides are probably COOH-terminally amidated by elimination of a glycine residue. These molecules with no evident homology to other hitherto described antimicrobial peptides were named penaeidins. PMID- 9353299 TI - HRX leukemic fusion proteins form a heterocomplex with the leukemia-associated protein SET and protein phosphatase 2A. AB - One of the most common chromosomal abnormalities in acute leukemia is a reciprocal translocation involving the HRX gene at chromosome locus 11q23, resulting in HRX fusion proteins. Using the yeast two-hybrid system, in vitro binding studies, and human cell culture coimmunoprecipitation experiments, we show here that a region of the HRX protein that is consistently retained in HRX leukemic fusion proteins interacts directly with SET, another protein implicated in leukemia. We have identified the binding sites on HRX for SET and show that these sequences are clustered near the A.T hooks that have been shown to bind DNA. We also show that carboxyl-terminal SET sequences, possibly the acidic tail of SET, bind to HRX. We have also found serine/threonine-specific protein phosphatase activity in anti-HRX coimmunoprecipitates. Using the phosphatase inhibitor okadaic acid and Western blotting, the phosphatase was identified as protein phosphatase 2A (PP2A). Mutation of a single amino acid in one of the SET binding sites of HRX resulted in lower amounts of both coimmunoprecipitated SET protein and coimmunoprecipitated PP2A. These results suggest that the leukemogenic effects of HRX fusion proteins may be related to interactions with SET and PP2A. PMID- 9353300 TI - Evidence that levels of presenilins (PS1 and PS2) are coordinately regulated by competition for limiting cellular factors. AB - Mutations in two related genes, PS1 and PS2, account for the majority of early onset cases of familial Alzheimer's disease. PS1 and PS2 are homologous polytopic membrane proteins that are processed endoproteolytically into two fragments in vivo. In the present report we examine the fate of endogenous PS1 and PS2 after overexpression of human PS1 or PS2 in mouse N2a neuroblastoma cell lines and human PS1 in transgenic mice. Remarkably, in N2a cell lines and in brains of transgenic mice expressing human PS1, accumulation of human PS1 derivatives is accompanied by a compensatory, and highly selective, decrease in the steady-state levels of murine PS1 and PS2 derivatives. Similarly, the levels of murine PS1 derivatives are diminished in cultured cells overexpressing human PS2. To define the minimal sequence requirements for "replacement" we expressed familial Alzheimer's disease-linked and experimental deletion variants of PS1. These studies revealed that compromised accumulation of murine PS1 and PS2 derivatives resulting from overexpression of human PS1 occurs in a manner independent of endoproteolytic cleavage. Our results are consistent with a model in which the abundance of PS1 and PS2 fragments is regulated coordinately by competition for limiting cellular factor(s). PMID- 9353301 TI - Molecular cloning and functional characterization of nitrobenzylthioinosine (NBMPR)-sensitive (es) and NBMPR-insensitive (ei) equilibrative nucleoside transporter proteins (rENT1 and rENT2) from rat tissues. AB - Equilibrative nucleoside transport processes in mammalian cells are either nitrobenzylthioinosine (NBMPR)-sensitive (es) or NBMPR-insensitive (ei). Previously, we isolated a cDNA from human placenta encoding the 456-residue glycoprotein hENT1. When expressed in Xenopus oocytes, hENT1 mediated es-type transport activity and was inhibited by coronary vasoactive drugs (dipyridamole and dilazep) that may compete with nucleosides and NBMPR for binding to the substrate binding site. We now report the molecular cloning and functional expression of es and ei homologs of hENT1 from rat tissues; rENT1 (457 residues) was 78% identical to hENT1 in amino acid sequence, and rENT2 (456 residues) was 49-50% identical to rENT1/hENT1 and corresponded to a full-length form of the delayed-early proliferative response gene product HNP36, a protein of unknown function previously cloned in truncated form. rENT1 was inhibited by NBMPR (IC50 = 4.6 nM at 10 microM uridine), whereas rENT2 was NBMPR-insensitive (IC50 > 1 microM). Both proteins mediated saturable uridine influx (Km = 0.15 and 0.30 mM, respectively), were broadly selective for purine and pyrimidine nucleosides, including adenosine, and were relatively insensitive to inhibition by dipyridamole and dilazep (IC50 > 1 microM). These observations demonstrate that es and ei nucleoside transport activities are mediated by separate, but homologous, proteins and establish a function for the HNP36 gene product. PMID- 9353302 TI - A stable alpha-helical domain at the N terminus of the RIalpha subunits of cAMP dependent protein kinase is a novel dimerization/docking motif. AB - The RIalpha subunit of cAMP-dependent protein kinase is maintained as an asymmetric dimer by a dimerization motif at the N terminus. Based on resistance to proteolysis and expression as a discrete domain in Escherichia coli, this motif is defined as residues 12-61. This motif is chemically, kinetically, and thermally stable. The two endogenous interchain disulfide bonds between Cys16 and Cys37 in RIalpha are extremely resistant to reduction even in 8 M urea, indicating that they are well shielded from the reducing environment of the cell. The disulfide bonds were present in recombinant RIalpha as well as when the dimerization domain alone was expressed in E. coli, emphasizing the unusual stability of this motif and the disulfide bonds. Although 100 mM dithiothreitol was sufficient to reduce the disulfide bonds, it did not abolish dimerization. In addition, a stable dimer also still formed when Cys37 was replaced with His, confirming unambiguously the original antiparallel alignment of the disulfide bonds. Thus, both in vitro and in vivo, disulfide bonds are not required for dimerization. Circular dichroism of the dimerization domain indicated a high content of a thermostable alpha-helix. Based on the CD data, trypsin resistance of the fragment, location of the disulfide bonds, and amphipathic helix predictions, potential models are discussed. A new alignment of the dimerization domains of RI, RII, and cGMP-dependent protein kinase elucidates fundamental similarities as well as significant differences among these three domains. PMID- 9353303 TI - Specificity of the ubiquitin isopeptidase in the PA700 regulatory complex of 26 S proteasomes. AB - The specificity of the ubiquitin (Ub) isopeptidase in the PA700 regulatory complex of the bovine 26 S proteasome was investigated. Disassembly of poly-Ub by this enzyme is restricted to the distal-end Ub of the substrate, i.e. the Ub farthest from the site of protein attachment in poly-Ub-protein conjugates. The determinants recognized by the isopeptidase were probed by the use of mutant ubiquitins incorporated into Lys48-linked poly-Ub substrates. PA700 could not disassemble poly-Ub chains that contained a distal Ub(L8A,I44A). This suggested either that the enzyme interacts directly with Leu8 or Ile44 or that it recognizes a higher order structure that caps the distal end of a poly-Ub substrate and is destabilized by Ub(L8A,I44A). The previously determined di-Ub crystal structure (Cook, W. J., Jeffrey, L. C., Carson, M., Chen, Z., and Pickart, C. M. (1992) J. Biol. Chem. 267, 16467-16471) offered a candidate for such a "cap." In solution, however, this structure was not observed by 1H NMR spectroscopy. This and the finding that di-Ub with a single proximal Ub(L8A,I44A) is cleaved efficiently suggest that Leu8 and Ile44 in the distal-end Ub contact the isopeptidase directly. In addition to Lys48-linked chains, PA700 also could disassemble Lys6- and Lys-11-linked poly-Ub, but, surprisingly, not alpha-linked di-Ub. Results with these and other substrates suggest that specificity determinants for the PA700 isopeptidase include Leu8, Ile44, and Lys48 on the distal Ub and, for poly-Ub, some features of the Ub-Ub linkage itself. PMID- 9353304 TI - Targeting of Tiam1 to the plasma membrane requires the cooperative function of the N-terminal pleckstrin homology domain and an adjacent protein interaction domain. AB - The Rho-like GTPases Cdc42, Rac, and Rho play key roles in the regulation of the actin cytoskeleton and are implicated in transcriptional activation and cell transformation. We have previously identified the invasion-inducing Tiam1 gene, which encodes an activator of Rac. In fibroblasts, Tiam1 induces Rac-mediated membrane ruffling, which requires the N-terminal pleckstrin homology (PHn) domain. Here we show that this PHn domain is part of a protein interaction domain, which mediates membrane localization of Tiam1. After subcellular fractionation, up to 50% of Tiam1 is recovered in the Triton X-100-insoluble high speed pellet that contains small protein complexes. The regions in Tiam1 that are responsible for these protein interactions comprise the PHn domain, an adjacent putative coiled coil region (CC), and an additional flanking region (Ex). Deletions in each of these regions abolish membrane localization of Tiam1 and membrane ruffling, suggesting that they function cooperatively. Indeed, only polypeptides encompassing the PHn-CC-Ex region, and not the PHn-CC or the Ex region, localize at the membrane. These results indicate that the N-terminal PH domain is part of a larger functional Tiam1 domain that mediates protein complex formation and membrane localization of Tiam1. PMID- 9353305 TI - Structure of nitrite bound to copper-containing nitrite reductase from Alcaligenes faecalis. Mechanistic implications. AB - The structures of oxidized, reduced, nitrite-soaked oxidized and nitrite-soaked reduced nitrite reductase from Alcaligenes faecalis have been determined at 1.8 2.0 A resolution using data collected at -160 degrees C. The active site at cryogenic temperature, as at room temperature, contains a tetrahedral type II copper site liganded by three histidines and a water molecule. The solvent site is empty when crystals are reduced with ascorbate. A fully occupied oxygen coordinate nitrite occupies the solvent site in crystals soaked in nitrite. Ascorbate-reduced crystals soaked in a glycerol-methanol solution and nitrite at 40 degrees C remain colorless at -160 degrees C but turn amber-brown when warmed, suggesting that NO is released. Nitrite is found at one-half occupancy. Five new solvent sites in the oxidized nitrite bound form exhibit defined but different occupancies in the other three forms. These results support a previously proposed mechanism by which nitrite is bound primarily by a single oxygen atom that is protonable, and after reduction and cleavage of that N-O bond, NO is released leaving the oxygen atom bound to the Cu site as hydroxide or water. PMID- 9353306 TI - Prion protein aggregation reverted by low temperature in transfected cells carrying a prion protein gene mutation. AB - Prion diseases are characterized by the conversion of the normal cellular prion protein (PrPC), a glycoprotein that is anchored to the cell membrane by a glycosylphosphatidylinositol moiety, into an isoform that is protease-resistant (PrPres) and pathogenic. In inherited prion diseases, mutations in the prion protein (PrPM) engender the conversion of PrPM into PrPres. We developed a cell model of Gerstmann-Straussler-Scheinker disease, a neurodegenerative condition characterized by PrPM-containing amyloid deposits and neuronal loss, by expressing the Gerstmann-Straussler-Scheinker haplotype Q217R-129V in human neuroblastoma cells. By comparison to PrPC, this genotype results in the following alterations of PrPM: 1) expression of an aberrant form lacking the glycosylphosphatidylinositol anchor, 2) increased aggregation and protease resistance, and 3) impaired transport to the cell surface. Most of these alterations are temperature-sensitive, indicating that they are due to misfolding of PrPM. PMID- 9353307 TI - The glucocorticoid receptor is associated with the RNA-binding nuclear matrix protein hnRNP U. AB - The glucocorticoid receptor (GR) is a ligand-dependent transcription factor that is able to modulate gene activity by binding to its response element, interacting with other transcription factors, and contacting several accessory proteins such as coactivators. Here we show that GRIP120, one of the factors we have identified to interact with the glucocorticoid receptor, is identical to the heterogeneous nuclear ribonucleoprotein U (hnRNP U), a nuclear matrix protein binding to RNA as well as to scaffold attachment regions. GR.hnRNP U complexes were identified by blotting and coimmunoprecipitation. The subnuclear distribution of GR and hnRNP U was characterized by indirect immunofluorescent labeling and confocal laser microscopy demonstrating a colocalization of both proteins. Using a nuclear transport-deficient deletion of hnRNP U, nuclear translocation was seen to be dependent on GR and dexamethasone. Transient transfections were used to identify possible interaction domains. Overexpressed hnRNP U interfered with glucocorticoid induction, and the COOH-terminal domains of both proteins were sufficient in mediating the transcriptional interference. A possible functional role for this GR binding-protein in addition to its binding to the nuclear matrix, to RNA, and to scaffold attachment regions is discussed. PMID- 9353309 TI - Characterization of two homologous yeast genes that encode mitochondrial iron transporters. AB - Two different yeast genes were identified that when overexpressed suppressed the low iron growth defect of a mutation in the endoplasmic reticulum iron binding enzyme methyl sterol oxidase. These genes were determined to be novel and highly related. The deduced amino acid sequences indicated that both were membrane proteins having two identical histidine-rich motifs. The predicted proteins, while not ABC transporters, are homologous to a widely distributed family of transition metal transporters present in all kingdoms. Subcellular fractionation and fluorescence microscopy localized these gene products to mitochondria. Based on this result we term these genes Mitochondrial Fe Transporters (MFT). Cells with disruptions in both genes show a growth defect on low iron medium, suggesting that these genes have redundant function and can affect cytosolic iron levels. Measurement of mitochondrial iron in cells grown in iron-rich medium overexpressing MFT1 or MFT2 show a 2-5-fold increase in iron compared with mitochondria from control cells. These results suggest that the mitochondria may act as a reservoir for iron that can be mobilized and used for cytosolic purposes. PMID- 9353308 TI - Regulation of cyclin D1 by calpain protease. AB - Cyclin D1, a critical positive regulator of G1 progression, has been implicated in the pathogenesis of certain cancers. Regulation of cyclin D1 occurs at the transcriptional and posttranscriptional level. Here we present evidence that cyclin D1 levels are regulated at the posttranscriptional level by the Ca2+ activated protease calpain. Serum starvation of NIH 3T3 cells resulted in rapid loss of cyclin D1 protein that was completely reversible by calpain inhibitors. Actinomycin D and lovastatin induced rapid loss of cyclin D1 in prostate and breast cancer cells that was reversible by calpain inhibitors and not by phenylmethylsulfonyl fluoride, caspase inhibitors, or lactacystin, a specific inhibitor of the 26 S proteasome. Treatment of intact NIH 3T3, prostate, and breast cancer cells with a calpain inhibitor dramatically increased the half-life of cyclin D1 protein. Addition of purified calpain to PC-3-M lysates resulted in Ca2+-dependent cyclin D1 degradation. Transient expression of the calpain inhibitor calpastatin increased cyclin D1 protein in serum-starved NIH 3T3 cells. Cyclins A, E, and B1 have been reported to be regulated by proteasome-associated proteolysis. The data presented here implicate calpain in cyclin D1 posttranslational regulation. PMID- 9353310 TI - Interaction of the second binding region of troponin I with the regulatory domain of skeletal muscle troponin C as determined by NMR spectroscopy. AB - Two dimensional 1H,15N-heteronuclear single quantum correlation NMR was used to monitor the resonance frequency changes of the backbone amide groups belonging to the 15N-labeled regulatory domain of calcium saturated troponin C (N-TnC) upon addition of synthetic skeletal N-acetyl-troponin I 115-131-amide peptide (TnI115 131). Utilizing the change in amide chemical shifts, the dissociation constant for 1:1 binding of TnI115-131 to N-TnC in low salt and 100 mM KCl samples was determined to be 28 +/- 4 and 24 +/- 4 microM, respectively. The off rate of TnI115-131 was determined to be 300 s-1 from observed N-TnC backbone amide 1H,15N heteronuclear single quantum correlation cross-peak line widths, which is on the order of the calcium off rates (Li, M. X., Gagne, S. M., Tsuda, S., Kay, C. M., Smillie, L. B., and Sykes, B. D. (1995) Biochemistry 34, 8330-8340), and agrees with kinetic expectations for biological regulation of muscle contraction. The TnI115-131 binding site on N-TnC was determined by mapping of chemical shift changes onto the N-TnC NMR structure and was demonstrated to be in the "hydrophobic pocket" (Gagne, S. M., Tsuda, S., Li, M. X., Smillie, L. B., and Sykes, B. D. (1995) Nat. Struct. Biol. 2, 784-789). PMID- 9353311 TI - Cell cycle-regulated expression, phosphorylation, and degradation of p55Cdc. A mammalian homolog of CDC20/Fizzy/slp1. AB - p55Cdc is a mammalian protein that shows high homology to the cell cycle proteins Cdc20p of Saccharomyces cerevisiae and the product of the Drosophila fizzy (fzy) gene, both of which contain WD repeats and are thought to be required for the metaphase-anaphase transition. The fzy mutants exhibit a metaphase arrest phenotype, which is accompanied by stabilization of cyclins A and B, leading to the hypothesis that fzy function is required for cell cycle-regulated ubiquitin mediated proteolysis. p55Cdc expression was initiated at the G1/S transition and steady state levels of p55Cdc were highest at M and lowest in G1. Inhibition of the 26 S proteasome prevented both mitotic exit and loss of p55Cdc at the M/G1 transition, suggesting that p55Cdc degradation was mediated by the cell cycle regulated proteolytic pathway. Immune complexes of p55Cdc obtained at different cell cycle stages showed a variety of proteins with dramatic differences observed in the pattern of associated proteins during the transition from G2 to M. Immunolocalization of p55Cdc demonstrated dynamic changes in p55Cdc localization as the cells transit mitosis. p55Cdc appears to act as a regulatory protein interacting with several other proteins, perhaps via its seven WD repeats, at multiple points in the cell cycle. PMID- 9353312 TI - Crystal structure of the I domain from integrin alpha2beta1. AB - We have determined the high resolution crystal structure of the I domain from the alpha-subunit of the integrin alpha2beta1, a cell surface adhesion receptor for collagen and the human pathogen echovirus-1. The domain, as expected, adopts the dinucleotide-binding fold, and contains a metal ion-dependent adhesion site motif with bound Mg2+ at the top of the beta-sheet. Comparison with the crystal structures of the leukocyte integrin I domains reveals a new helix (the C-helix) protruding from the metal ion-dependent adhesion site face of the domain which creates a groove centered on the magnesium ion. Modeling of a collagen triple helix into the groove suggests that a glutamic acid side chain from collagen can coordinate the metal ion, and that the C-helix insert is a major determinant of binding specificity. The binding site for echovirus-1 maps to a distinct surface of the alpha2-I domain (one edge of the beta-sheet), consistent with data showing that virus and collagen binding occur by different mechanisms. Comparison with the homologous von Willebrand factor A3 domain, which also binds collagen, suggests that the two domains bind collagen in different ways. PMID- 9353314 TI - The 5'-exonuclease activity of bacteriophage T4 RNase H is stimulated by the T4 gene 32 single-stranded DNA-binding protein, but its flap endonuclease is inhibited. AB - Bacteriophage T4 RNase H is a 5'- to 3'-nuclease that has exonuclease activity on RNA.DNA and DNA.DNA duplexes and can remove the pentamer RNA primers made by the T4 primase-helicase (Hollingsworth, H. C., and Nossal, N. G. (1991) J. Biol. Chem. 266, 1888-1897; Hobbs, L. J., and Nossal, N. G. (1996) J. Bacteriol. 178, 6772-6777). Here we show that this exonuclease degrades duplex DNA nonprocessively, releasing a single oligonucleotide (nucleotides 1-4) with each interaction with the substrate. Degradation continues nonprocessively until the enzyme stops 8-11 nucleotides from the 3'-end of the substrate. T4 gene 32 single stranded DNA-binding protein strongly stimulates the exonuclease activity of T4 RNase H, converting it into a processive nuclease that removes multiple short oligonucleotides with a combined length of 10-50 nucleotides each time it binds to the duplex substrate. 32 protein must bind on single-stranded DNA behind T4 RNase H for processive degradation. T4 RNase H also has a flap endonuclease activity that cuts preferentially on either side of the junction between single- and double-stranded DNA in flap and fork DNA structures. In contrast to the exonuclease, the endonuclease is inhibited completely by 32 protein binding to the single strand of the flap substrate. These results suggest an important role for T4 32 protein in controlling T4 RNase H degradation of RNA primers and adjacent DNA during each lagging strand cycle. PMID- 9353313 TI - Echovirus 1 interaction with the human very late antigen-2 (integrin alpha2beta1) I domain. Identification of two independent virus contact sites distinct from the metal ion-dependent adhesion site. AB - The human integrin very late antigen (VLA)-2 (CD49b/CD29) mediates interactions with collagen and is the receptor for echovirus 1. Binding sites for both collagen and echovirus 1 have been mapped to the I domain within the alpha2 subunit of the VLA-2 alpha2beta1 heterodimer. Although murine VLA-2 interacts with collagen, it does not bind virus. We have used isolated human-murine chimeric I domains expressed as glutathione S-transferase fusion proteins in Escherichia coli to identify two groups of amino acids, 199-201 and 212-216, independently involved in virus attachment. These residues are distinct from the metal ion-dependent adhesion site previously demonstrated to be essential for VLA 2 interactions with collagen. Mutations in three metal ion-dependent adhesion site residues that abolish adhesion to collagen had no effect on virus binding. These results confirm that different sites within the I domain are responsible for VLA-2 interaction with extracellular matrix proteins and with viral ligands. PMID- 9353315 TI - Identification of residues of T4 RNase H required for catalysis and DNA binding. AB - Bacteriophage T4 RNase H, which removes the RNA primers that initiate lagging strand fragments, has a 5'- to 3'-exonuclease activity on DNA.DNA and RNA.DNA duplexes and an endonuclease activity on flap or forked DNA structures (Bhagwat, M., Hobbs, L. J., and Nossal, N. J. (1997) J. Biol. Chem. 272, 28523-28530). It is a member of the RAD2 family of prokaryotic and eukaryotic replication and repair nucleases. The crystal structure of T4 RNase H, in the absence of DNA, shows two Mg2+ ions coordinated to the amino acids highly conserved in this family. It also shows a disordered region proposed to be involved in DNA binding (Mueser, T. C., Nossal, N. G., and Hyde, C. C. Cell (1996) 85, 1101-1112). To identify the amino acids essential for catalysis and DNA binding, we have constructed and characterized three kinds of T4 RNase H mutant proteins based on the possible roles of the amino acid residues: mutants of acidic residues coordinated to each of the two Mg2+ ions (Mg2+-1: D19N, D71N, D132N, and D155N; and Mg2+-2: D157N and D200N); mutants of conserved basic residues in or near the disordered region (K87A and R90A); and mutants of residues with hydroxyl side chains involved in the hydrogen bonding network (Y86F and S153A). Our studies show that Mg2+-1 and the residues surrounding it are important for catalysis and that Lys87 is necessary for DNA binding. PMID- 9353317 TI - Deletions of the Aequorea victoria green fluorescent protein define the minimal domain required for fluorescence. AB - The Green Fluorescent Protein (GFP) from the jellyfish Aequorea victoria is a widely used marker for gene expression and protein localization studies. Dissection of the structure of the protein would be expected to shed light on its potential applications to other fields such as the detection of protease activity. Using deletion analysis, we have defined the minimal domain in GFP required for fluorescence to amino acids 7-229. This domain starts at the middle of the first small alpha helix at the N terminus of GFP and ends immediately following the last beta sheet. Studies of the amino acids at both termini of the minimal domain revealed that positions 6 and 7 at the N terminus are Glu specific. Change of the Glu residues to other amino acids results in reduction of GFP fluorescence. Position 229 at the C terminus of GFP, however, is nonspecific: the Ile can be replaced with other amino acids with no measurable loss of fluorescence. A total of only 15 terminal amino acids can be deleted from GFP without disrupting fluorescence, consistent with findings of a previous study of GFP crystal structure (Ormo, M., Cubitt, A. B., Kallio, K., Gross, L. A., Tsien, R. Y., Remington, S. J. (1996) Science 273, 1392-1395 and Yang, F., Moss, L. G., and Phillips, G. N., Jr. (1996) Nat. Biotechnol. 14, 1246-1251) that a tightly packed structure exists in the protein. We also generated internal deletions within the loop regions of GFP according to its crystal structure and found that all such deletions eliminated GFP fluorescence. PMID- 9353316 TI - Purification and biochemical properties of Saccharomyces cerevisiae Mdj1p, the mitochondrial DnaJ homologue. AB - The DnaK/DnaJ/GrpE heat shock proteins of Escherichia coli constitute the prototype DnaK chaperone machine. Various studies have shown that these three proteins work synergistically in a diverse array of biological functions, including protein folding and disaggregation, proteolysis, and transport across biological membranes. We have overexpressed and purified the mitochondrial Saccharomyces cerevisiae DnaJ homologue, Mdj1pDelta55, which lacks the mitochondrial presequence, and studied its biochemical properties in well defined in vitro systems. We find that Mdj1pDelta55 interacts with DnaK as judged both by an enzyme-linked immunosorbent assay, as well as stimulation of DnaK's weak ATPase activity in the presence of GrpE. In addition, Mdj1pDelta55 not only interacts with denatured firefly luciferase on its own, but also enables DnaK to bind to it in an ATP-dependent mode. Using co-immunoprecipitation assays we can demonstrate the presence of a stable Mdj1pDelta55-luciferase-DnaK complex. However, in contrast to DnaJ, Mdj1pDelta55 does not appear to interact well with certain seemingly folded proteins, such as the sigma32 heat shock transcription factor or the lambdaP DNA replication protein. Finally, Mdj1pDelta55 can substitute perfectly well for DnaJ in the refolding of denatured firefly luciferase by the DnaK chaperone machine. These studies demonstrate that Mdj1pDelta55 has conserved most of DnaJ's known biological properties, thus supporting an analogous functional role in yeast mitochondria. PMID- 9353318 TI - The chicken GATA-6 locus contains multiple control regions that confer distinct patterns of heart region-specific expression in transgenic mouse embryos. AB - The GATA-6 transcription factor is expressed in cardiogenic cells and during subsequent stages of heart development in diverse vertebrate species. To gain insights into the molecular events that govern this heart-restricted expression, we isolated the chicken GATA-6 gene and used several approaches to screen for associated control regions. Our analysis of two chicken GATA-6/lacZ constructs in transgenic mouse embryos was particularly revealing. One GATA-6/lacZ construct, which has 1.5 kilobase pairs of upstream sequences along with the promoter and first intron, was expressed exclusively in the atrioventricular canal region of the heart. This expression pattern is novel and appears to mark specialized myocardial cells that induce underlying endocardial cells to initiate valve formation. The other GATA-6/lacZ construct, which has an additional 7.7 kilobase pairs of upstream sequences, was expressed in the ventricle and outflow tract in addition to the atrioventricular canal. The failure of these GATA-6 control regions to function as enhancers in transfected cardiac myocyte cultures underscores the importance of using transgenic approaches to elucidate transcriptional controls that function in the developing heart. Although the endogenous GATA-6 gene is expressed throughout the heart, our results indicate that this is effected in a heart region-specific manner. PMID- 9353319 TI - Characterization of NEDD8, a developmentally down-regulated ubiquitin-like protein. AB - NEDD8 is a novel 81 amino acid polypeptide which is 60% identical and 80% homologous to ubiquitin. Northern blot analysis showed that the NEDD8 message was developmentally down-regulated. In adult tissues, NEDD8 expression was mostly restricted to the heart and skeletal muscle. Antiserum specific for NEDD8 detected a 6-kDa monomer in SK-N-SH, BJAB, and HL60 cell lysates. A 14-kDa band was also detected in BJAB, HL60, and SK-MEL28 but not in SK-N-SH and K562 cell lysates. An approximately 90-kDa band was detected in all cell lines tested. Thus, NEDD8 is likely to be conjugated to other proteins in a manner analogous to ubiquitination. However, the conjugation pattern of NEDD8 is entirely different from that of ubiquitin in all cell lines tested. To study NEDD8 conjugation in more detail, hemagglutinin-epitope-tagged NEDD8 was expressed in COS cells. Western blot analysis revealed an NEDD8 monomer and a series of higher molecular weight NEDD8-conjugated proteins or NEDD8 multimers. Immunocytochemical analysis showed that NEDD8 expression was highly enriched in the nucleus and was much weaker in the cytosol. In contrast, ubiquitin expression was detectable equally well in the nucleus and cytosol. Mutational analysis showed that the C terminus of NEDD8 was efficiently cleaved and that Gly-76 was required for conjugation of NEDD8 to other proteins. Taken together, NEDD8 provides another substrate for covalent protein modification and may play a unique role during development. PMID- 9353320 TI - Urokinase receptor is associated with the components of the JAK1/STAT1 signaling pathway and leads to activation of this pathway upon receptor clustering in the human kidney epithelial tumor cell line TCL-598. AB - The urokinase-type plasminogen activator (uPA) binds to cells via a specific receptor attached to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. Despite the lack of a transmembrane domain, the urokinase receptor (uPAR) is capable of transducing extracellular signals affecting growth, migration, and adhesion. Several Tyr kinases of the src family as well as beta1, beta2, and beta3 integrins were found to be associated with the uPAR. We found that in the human kidney epithelial line TCL-598, also components of the JAK1/STAT1 signal transduction pathway including gp130, are associated with uPAR as revealed by coimmunoprecipitation and are co-localized in caveolae. Upon clustering of uPA.uPAR complex by a monoclonal antibody, JAK1 associates with uPAR, which in turn leads to STAT1 phosphorylation, dimerization, specific binding to DNA, and gene activation. To prove the dependence of STAT1 activation on the uPAR, TCL-598 cells were treated with sense and antisense uPAR oligonucleotides. In antisense treated cells in which uPAR expression was reduced to less then one third, activation of STAT1 by the clustering antibody was abolished while STAT1 activation by interferon-gamma was unaffected. Therefore, in this cell line, uPA.uPAR also utilizes the JAK1/STAT1 pathway for signaling, and gp130 might be the transmembrane adapter for this signal transduction pathway. PMID- 9353321 TI - Tissue factor is induced by monocyte chemoattractant protein-1 in human aortic smooth muscle and THP-1 cells. AB - Monocyte chemoattractant protein-1 (MCP-1) is a C-C chemokine thought to play a major role in recruiting monocytes to the atherosclerotic plaque. Tissue factor (TF), the initiator of coagulation, is found in the atherosclerotic plaque, macrophages, and human aortic smooth muscle cells (SMC). The exposure of TF during plaque rupture likely induces acute thrombosis, leading to myocardial infarction and stroke. This report demonstrates that MCP-1 induces the accumulation of TF mRNA and protein in SMC and in THP-1 myelomonocytic leukemia cells. MCP-1 also induces TF activity on the surface of human SMC. The induction of TF by MCP-1 in SMC is inhibited by pertussis toxin, suggesting that the SMC MCP-1 receptor is coupled to a Gi-protein. Chelation of intracellular calcium and inhibition of protein kinase C block the induction of TF by MCP-1, suggesting that in SMC it is mediated by activation of phospholipase C. SMC bind MCP-1 with a Kd similar to that previously reported for macrophages. However, mRNA encoding the macrophage MCP-1 receptors, CCR2A and B, is not present in SMC, indicating that they possess a distinct MCP-1 receptor. These data suggest that in addition to being a chemoattractant, MCP-1 may have a procoagulant function and raise the possibility of an autocrine pathway in which MCP-1, secreted by SMC and macrophages, induces TF activity in these same cells. PMID- 9353322 TI - Cytokeratin 18 is expressed on the hepatocyte plasma membrane surface and interacts with thrombin-antithrombin complexes. AB - During experiments to identify putative hepatic receptors for thrombin antithrombin (TAT) complexes, a 45-kDa protein was identified by ligand blotting. Following gel purification, amino acid sequencing revealed the 45-kDa TAT-binding polypeptide to be cytokeratin 18 (CK18). The presence of CK18 on the surface of intact rat hepatoma cells was demonstrated by binding of 125I-anti-CK18 antibodies. Anti-CK18 antibodies reduced the binding and internalization of 125I TAT by rat hepatoma cells. Immunocytochemical analysis, to determine the location of CK18 in vivo, revealed a periportal gradient of CK18 staining; with hepatocytes around the portal triads demonstrating striking pericellular staining. In addition, anti-CK18 IgG associated with perfused livers to a significantly greater extent than preimmune IgG. Taken together, these data provide evidence that CK18 is found on the extracellular surface of hepatocytes and could play a role in TAT removal. Finally, these data, in conjunction with recent reports of CK8 (Hembrough, T. A., Li, L., and Gonias, S. L. (1996) J. Biol. Chem. 271, 25684-25691) and CK1 cell membrane surface expression (Schmaier, A. H. (1997) Thromb. Hemostasis 78, 101-107), indicate a novel role for these proteins as putative cellular receptors or cofactors to cellular receptors. PMID- 9353324 TI - Presence of laminin alpha5 chain and lack of laminin alpha1 chain during human muscle development and in muscular dystrophies. AB - There is currently a great interest in identifying laminin isoforms expressed in developing and regenerating skeletal muscle. Laminin alpha1 has been reported to localize to human fetal muscle and to be induced in muscular dystrophies based on immunohistochemistry with the monoclonal antibody 4C7, suggested to recognize the human laminin alpha1 chain. Nevertheless, there seems to be no expression of laminin alpha1 protein or mRNA in developing or dystrophic mouse skeletal muscle fibers. To address the discrepancy between the results obtained in developing and dystrophic human and mouse muscle we expressed the E3 domain of human laminin alpha1 chain as a recombinant protein and made antibodies specific for human laminin alpha1 chain (anti-hLN-alpha1G4/G5). We also made antibodies to the human laminin alpha5 chain purified from placenta. In the present report we show that hLN-alpha1G4/G5 antibodies react with a 400-kDa laminin alpha1 chain and that 4C7 reacts with a 380-kDa laminin alpha5 chain. Immunohistochemistry with the hLN alpha1G4/G5 antibody and 4C7 revealed that the two antibodies stained human kidney, developing and dystrophic muscle in distinct patterns. Our data indicate that the previously reported expression patterns in developing, adult, and dystrophic human muscle tissues with 4C7 should be re-interpreted as an expression of laminin alpha5 chain. Our data are also consistent with earlier work in mouse, indicating that laminin alpha1 is largely an epithelial laminin chain not present in developing or dystrophic muscle fibers. PMID- 9353323 TI - Targeted inhibition of interferon-gamma-dependent intercellular adhesion molecule 1 (ICAM-1) expression using dominant-negative Stat1. AB - A subset of epithelial immune-response genes (including intercellular adhesion molecule-1 (ICAM-1)) depends on an IFN-gamma signal transduction pathway with the Stat1 transcription factor as a critical intermediate. Excessive local activation of this pathway may lead to airway inflammation, so we sought to selectively down regulate the pathway using a dominant-negative strategy for inhibition of epithelial Stat1 in a primary culture airway epithelial cell model. Using a Stat1 deficient cell line, we demonstrated that transfection of wild-type Stat1 expression plasmid restored appropriate Stat1 expression and IFN-gamma-dependent phosphorylation as well as consequent IFN-gamma activation of cotransfected ICAM 1 promoter constructs and endogenous ICAM-1 gene expression. However, mutations of Stat1 at Tyr-701 (JAK kinase phosphorylation site), Glu-428/429 (putative DNA binding site), His-713 (splice site resulting in Stat1beta formation), or Ser-727 (MAP kinase phosphorylation site) all decreased Stat1 capacity to activate the ICAM-1 promoter. The Tyr-701 mutant (followed by the His-713 mutant) were most effective in disabling Stat1 function and in overcoming the activating effect of cotransfected wild-type Stat1 in this cell system thereby highlighting the effectiveness of blocking Stat1 homo- and hetero-dimerization. In experiments using primary culture human tracheobronchial epithelial cells (hTBECs) and each of the four Stat1 mutant plasmids, transfection with the Tyr-701 and His-713 mutants again most effectively inhibited IFN-gamma activation of an ICAM-1 gene promoter construct. Then by transfecting hTBECs with wild-type or mutant Stat1 tagged with a Flag reporter sequence, we used dual immunofluorescence to show that hTBECs expressing the Tyr-701 or His-713 mutants were prevented from expressing endogenous ICAM-1 in response to IFN-gamma treatment. The capacity of a specific Stat1 mutations to exert a potent dominant-negative effect on IFN gamma signal transduction provides for further definition of Stat1 structure function and a means for natural or engineered expression of mutant Stat1 to selectively down-regulate activity of this pathway in a cell type- or tissue specific manner during immune and/or inflammatory responses. PMID- 9353325 TI - Mismatch repair defects and O6-methylguanine-DNA methyltransferase expression in acquired resistance to methylating agents in human cells. AB - Fifteen variants with >/=30-fold resistance to N-methyl-N-nitrosourea were isolated from the Burkitt's lymphoma Raji cell line. Eight had received a single treatment with a highly cytotoxic dose. The remainder, including the previously described RajiF12 cell line, arose following multiple exposures to initially moderate but escalating doses. Surprisingly, methylation resistance arose in three clones by reactivation of a previously silent O6-methylguanine-DNA methyltransferase gene. Five clones, including RajiF12, displayed the microsatellite instability and increased spontaneous mutation rates at the hypoxanthine-guanine phosphoribosyltransferase locus, consistent with deficiencies in mismatch repair. Defects in either the hMutSalpha or hMutLalpha mismatch repair complexes were identified in extracts of these resistant clones by in vitro complementation using extracts from colorectal carcinoma cell lines. Defects in hMutLalpha were confirmed by Western blot analysis. Remarkably, five methylation-resistant clones in which mismatch repair defects were demonstrated by biochemical assays did not exhibit significant microsatellite instability. PMID- 9353326 TI - Hyperphosphorylation of the N-terminal domain of Cdc25 regulates activity toward cyclin B1/Cdc2 but not cyclin A/Cdk2. AB - Cdc25 regulates entry into mitosis by regulating the activation of cyclin B/cdc2. In humans, at least two cdc25 isoforms have roles in controlling the G2/M transition. Here we show, using bacterially expressed recombinant proteins, that two cdc25B splice variants, cdc25B2 and cdc25B3, are capable of activating cyclin A/cdk2 and cyclin B/cdc2, but that mitotic hyperphosphorylation of these proteins increases their activity toward only cyclin B1/cdc2. Cdc25C has only very low activity in its unphosphorylated form, and following hyperphosphorylation it will efficiently catalyze the activation of only cyclin B/cdc2. This was reflected by the in vivo activity of the immunoprecipitated cdc25B and cdc25C from interphase and mitotic HeLa cells. The increased activity of the hyperphosphorylated cdc25s toward cyclin B1/cdc2 was in large part due to increased binding of this substrate. The substrate specificity, activities, and timing of the hyperphosphorylation of cdc25B and cdc25C during G2 and M suggest that these two mitotic cdc25 isoforms are activated by different kinases and perform different functions during progression through G2 into mitosis. PMID- 9353327 TI - A peptidyl-prolyl cis/trans-isomerase (cyclophilin G) in regulated secretory granules. AB - A 27-kDa protein (p27) in horseshoe crab hemocyte that cross-reacts with antiserum against a beta-glucan-sensitive protease zymogen was purified to homogeneity, and its cDNA was cloned. The 1.7-kilobase pair cDNA contains an open reading frame of 660 base pairs, encoding a 23-amino acid signal sequence followed by a mature protein of 197 residues. The sequence of p27 exhibits strong similarity to that of cyclophilin B, a peptidyl-prolyl cis/trans-isomerase. p27 exhibits isomerase activity with a kcat/Km of 0.18 microM-1 s-1 for a peptide substrate; this activity is inhibited by cyclosporin A but is not affected by FK506. Although the p27 precursor possesses an amino-terminal secretory hydrophobic signal sequence, unlike other cyclophilin B molecules, it lacks a conserved carboxyl-terminal endoplasmic reticulum retention signal and it contains a central 8-amino acid insertion. Although p27 is secreted into the culture media of transiently expressed COS cells, it is not detected in horseshoe crab hemolymph plasma but rather is localized to the hemocyte large granules, the regulated secretory granules that are exocytosed upon stimulation. These results indicate that p27 is a new peptidyl-prolyl cis/trans-isomerase in the regulated secretory granules, and is thus designated cyclophilin G. This first report of a cyclophilin homologue in the secretory granule of the horseshoe crab hemocyte suggests that such chaperon-like proteins may constitute a key quality control system for stored proteins in exocytotic granules. PMID- 9353328 TI - Molecular cloning and functional expression of a cDNA encoding a new member of mixed lineage protein kinase from human brain. AB - We have cloned a novel protein kinase from human cerebellum and named it LZK (leucine zipper-bearing kinase). The LZK cDNA encoded a 966-amino acid polypeptide that contains a kinase catalytic domain and double leucine/isoleucine zippers separated by a short spacer region. The amino acid sequence of the kinase catalytic domain was a hybrid between those in serine/threonine and tyrosine protein kinases, indicating that LZK belongs to the subfamily of the mixed lineage kinase (MLK) family. The kinase catalytic domain of LZK was most similar to DLK (Holtzman, L. B., Merritt, S.E., and Fan, G. (1994) J. Biol. Chem. 269, 30808-30817), MUK (Hirai, S., Izawa, M., Osada, S., Spyrou, G., and Ohno, S. (1996) Oncogene 12, 641-650), and ZPK (Reddy, U. R., and Presure, D. (1994) Biochem. Biophys. Res. Commun. 202, 613-620), which belong to the same subfamily of the MLK family. However, besides the kinase catalytic domain and double leucine/isoleucine zippers, there was no significant homology with known proteins. The recombinant LZK autophosphorylated in the presence of ATP and divalent cations, and exhibited serine/threonine kinase catalytic activity. Northern blot analysis revealed that LZK is expressed most strongly in the pancreas, with a pattern that differs from other MLKs. Expression of LZK in COS7 cells induced phosphorylation of c-Jun and activation of JNK-1, indicating the association of LZK in the c-Jun amino-terminal kinase/stress-activated protein kinase pathway. The expressed LZK was detected primarily in the membrane fraction, suggesting that LZK interacts with other cellular components in vivo. PMID- 9353329 TI - Immunological and biological properties of Bet v 4, a novel birch pollen allergen with two EF-hand calcium-binding domains. AB - We have isolated a cDNA clone coding for a birch pollen allergen, Bet v 4. The deduced amino acid sequence of Bet v 4 contained two typical EF-hand calcium binding domains. Sequence similarities of Bet v 4 to calmodulin are primarily confined to the calcium-binding domains. However, significant sequence similarities extending outside the Ca2+-binding sites were found with a recently described group of pollen-specific allergens of Brassica and Bermuda grass. Both EF-hand domains of Bet v 4 are able to bind Ca2+, as demonstrated by 45Ca2+ blot overlay of wild type and calcium-binding deficient mutants of Bet v 4. Among pollen-allergic patients, protein-bound Ca2+ was not an absolute requirement for IgE recognition of Bet v 4. However, disruption of the carboxyl-terminal Ca2+ binding domain indicated that most IgE antibodies from allergic patients are directed against this site. IgE inhibition experiments suggested that Bet v 4 represents a highly cross-reactive pollen allergen. Pre-absorption of allergic sera with Bet v 4 drastically reduced IgE binding to proteins of similar molecular weight in pollen extracts from distantly related plant species (e.g. timothy grass, mugwort, lily) but not in extracts from plant-derived foodstuff. To test for a possible biological role in pollen germination and tube growth, we introduced recombinant Bet v 4 protein into growing lily pollen tubes by iontophoresis. As a result, cytoplasmic streaming stopped in the vicinity of the electrode tip, and a slight depolarization of the membrane voltage was measured. These effects were not observed with Ca2+-binding deficient mutants of Bet v 4. Thus, Bet v 4 and homologous proteins represent a new class of pollen-specific Ca2+-binding allergens that may have a physiological role as inhibitors of cytoplasmic streaming in outgrowing pollen tubes. PMID- 9353330 TI - UDP-GlcNAc:Ser-protein N-acetylglucosamine-1-phosphotransferase from Dictyostelium discoideum recognizes serine-containing peptides and eukaryotic cysteine proteinases. AB - Phosphoglycosylation catalyzed by UDP-GlcNAc:Ser-protein N-acetylglucosamine-1 phosphotransferase (Ser:GlcNAc phosphotransferase) adds GlcNAcalpha-1-P to peptidyl-Ser of selected Dictyostelium discoideum proteins. Lysosomal cysteine proteinase (CP), proteinase-1(CP7), is the major phosphoglycosylated protein in bacterially grown amoebae. GlcNAc-1-P is added within a Ser-rich domain containing SSS, SGSG, or SGSQ repeated motifs that are not found in other papain like CPs. We studied the substrate specificity of the transferase using peptides containing these motifs and 12 other peptides with one or more Ser residues. Phosphoglycosylation is comparable for all three Dictyostelium CP motifs, but it is not restricted to them. Flanking residues in the other peptides strongly influence phosphoglycosylation efficiency. Dictyostelium microsomal membranes also phosphoglycosylate endogenous acceptors, and some of these acceptors occur as an 18 S complex with the transferase. CP-serine motif peptides inhibit endogenous acceptor phosphoglycosylation weakly (30-40%) at 800 microM, whereas catalytically inactive proteinase-1(CP7) and other non-phosphoglycosylated eukaryotic CPs, lacking the serine domain, inhibit transferase activity at 1-4 microM. SDS denaturation destroys the inhibitory potential of all CPs showing that transferase recognizes a conformation-dependent feature that is shared by all. Proteinase-1(CP7) expressed in Escherichia coli lacks GlcNAc-1-P, but it is a substrate for Ser:GlcNAc phosphotransferase, Km = 5.6 microM. Thus, Ser:GlcNAc phosphotransferase recognizes both acceptor peptide sequences and a conformational feature of eukaryotic CPs. This may be physiologically important for establishing or maintaining non-overlapping groups of GlcNAc-1-P- and Man-6-P modified Dictyostelium proteins that reside in functionally distinct endo lysosomal vesicles. PMID- 9353331 TI - Human Prk is a conserved protein serine/threonine kinase involved in regulating M phase functions. AB - Human prk encodes a novel protein serine/threonine kinase capable of strongly phosphorylating casein but not histone H1 in vitro. prk expression is tightly regulated at various levels during different stages of the cell cycle in lung fibroblasts. The Prk kinase activity is relatively low during mitosis, G1, and G1/S, and peaks during late S and G2 stages of the cell cycle. Recombinant human Prk expressed through the baculoviral vector system is capable of phosphorylating Cdc25C, a positive regulator for the G2/M transition. Human prk shares significant sequence homology with Saccharomyces cerevisiae CDC5 and Drosophila melanogaster polo, both of which are essential for mitosis and meiosis. Full length prk transcripts greatly potentiate progesterone-induced meiotic maturation of Xenopus laevis oocytes. On the other hand, antisense prk transcripts significantly delay and reduce the rate of oocyte maturation. When expressed in a CDC5 mutant strain of S. cerevisiae, human Prk, but not a deletional mutant protein, fully rescues the temperature-sensitive phenotype of the budding yeast. Taken together, prk may represent a new protein kinase, playing an important role in regulating the onset and/or progression of mitosis in mammalian cells. PMID- 9353332 TI - Cloning, expression, and characterization of two manganese superoxide dismutases from Caenorhabditis elegans. AB - Two genes encoding manganese superoxide dismutase (sod-2 and sod-3) have been identified in the nematode Caenorhabditis elegans. Each gene is composed of five exons, and intron positions are identical; however, intron sizes and sequences are not the same. The predicted protein sequences are 86.3% homologous (91.8% conservative), and the cDNAs are only 75.2% homologous. Both deduced protein sequences contain the expected N-terminal mitochondrial transit peptides. Reverse transcriptase polymerase chain reaction analysis shows that both genes are expressed under normal growth conditions and that their RNA transcripts are trans spliced to the SL-1 leader sequence. The latter result together with Northern blot analysis indicate that both genes have mono-cistronic transcripts. The sod-3 gene was mapped to chromosome X, and the location of sod-2 was confirmed to be chromosome I. Polymerase chain reaction was used to amplify the cDNA regions encoding the predicted mature manganese superoxide dismutase proteins and each was cloned and expressed to high levels in Escherichia coli cells deficient in cytosolic superoxide dismutases. Both proteins were shown to be active in E. coli, providing similar protection against methyl viologen-induced oxidative stress. The expressed enzymes, which were not inhibited by hydrogen peroxide or cyanide, are dimeric, show quite different electrophoretic mobilities and isoelectric points, but exhibit comparable specific activities. PMID- 9353334 TI - Modified low density lipoprotein enhances the secretion of bile salt-stimulated cholesterol esterase by human monocyte-macrophages. species-specific difference in macrophage cholesteryl ester hydrolase. AB - Reverse transcriptase-polymerase chain reaction was used to study the biosynthesis of two different cholesteryl ester hydrolases by human and mouse macrophages. Oligonucleotide primers for bile salt-stimulated cholesterol esterase yielded positive reactions with RNA isolated from human peripheral blood monocytes, monocyte-derived macrophages, the human monocytic THP-1 cells, and phorbol ester-induced THP-1 macrophages. In contrast, oligonucleotide primers for hormone-sensitive lipase yielded positive reactions only with RNA isolated from non-differentiated human THP-1 monocytic cells and peripheral blood monocytes, but not those obtained from differentiated THP-1 macrophages or monocyte-derived macrophages. Thus, while human monocytes were capable of synthesizing both enzymes, human macrophages synthesized only bile salt-stimulated cholesterol esterase and not the hormone-sensitive lipase. The synthesis of bile salt stimulated cholesterol esterase by human macrophages was confirmed by detection of bile salt-stimulated cholesteryl ester hydrolytic activity in conditioned media of differentiated THP-1 cells and human peripheral blood monocyte-derived macrophages. Moreover, incubating human macrophages with oxidized low density lipoprotein (LDL) or acetylated LDL increased bile salt-stimulated cholesterol esterase activity in the conditioned media of these cells. These results with human macrophages were contrasted with results of studies with mouse macrophages, which showed the presence of hormone-sensitive lipase mRNA but not the bile salt stimulated cholesterol esterase mRNA. Taken together, these results demonstrated species-specific differences in expression of cholesteryl ester hydrolytic enzymes in macrophages. The expression of bile salt-stimulated cholesterol esterase by human macrophages, in a process inducible by modified LDL, suggests a role of this protein in atherogenesis. PMID- 9353333 TI - Inhibition of meizothrombin and meizothrombin(desF1) by heparin cofactor II. AB - Meizothrombin and meizothrombin(desF1) are intermediates formed during the conversion of prothrombin to thrombin by factor Xa, factor Va, phospholipids, and Ca2+ (prothrombinase). These intermediates are active toward synthetic peptide substrates but have limited ability to interact with platelets or macromolecular substrates such as fibrinogen. Meizothrombin and meizothrombin(desF1) activate protein C, however, and may exert primarily an anticoagulant effect. In this study, we investigated the inhibition of meizothrombin and meizothrombin(desF1) by two glycosaminoglycan-dependent protease inhibitors, heparin cofactor II (HCII) and antithrombin (AT). Purified recombinant meizothrombin and meizothrombin(desF1) were inhibited by HCII in the presence of dermatan sulfate with maximal second-order rate constants of 8 x 10(6) M-1.min-1 and 1.8 x 10(7) M 1.min-1, respectively, but were inhibited less than one-tenth as fast by AT in the presence of heparin. Similarly, the products of the prothrombinase reaction were inhibited in situ more effectively by HCII than by AT. When HCII and dermatan sulfate were present continuously during the prothrombinase reaction, meizothrombin was trapped as a sodium dodecyl sulfate-stable complex with HCII and no amidolytic activity could be detected with a thrombin substrate. Our findings indicate that HCII is an effective inhibitor of meizothrombin and meizothrombin(desF1) and, therefore, might regulate the anticoagulant activity of these proteases. PMID- 9353335 TI - Identification of sites required for down-regulation of Na+/H+ exchanger NHE3 activity by cAMP-dependent protein kinase. phosphorylation-dependent and independent mechanisms. AB - We recently identified a region within the cytoplasmic C-terminal tail of the Na+/H+ exchanger NHE3 isoform (residues 579 to 684) which is essential for inhibition of transport activity by cAMP-dependent protein kinase (PKA) (Cabado, A. G., Yu, F. H., Kapus, A., Gergely, L., Grinstein, S., and Orlowski, J. (1996) J. Biol. Chem. 271, 3590-3599). To further define determinants of PKA regulation, six serine residues located in potential recognition sequences for PKA within, or adjacent to, this region (positions 552, 605, 634, 661, 690, and 691) were altered either independently or in various combinations using site-directed mutagenesis. Wild type and mutant NHE3s tagged with the influenza virus hemagglutinin epitope were stably expressed in exchanger-deficient Chinese hamster ovary cells (AP-1) for functional studies. Of the individual mutations examined, only substitutions at Ser605 or Ser634 affected sensitivity to forskolin, an activator of adenylate cyclase, although partial inhibition of NHE3 activity by forskolin remained. By contrast, simultaneous mutation of both these serines completely abolished cAMP-mediated inhibition of NHE3 without greatly affecting basal transport activity. Two-dimensional analysis of tryptic digests of immunoprecipitated NHE3 labeled in vivo with [32P]orthophosphate revealed several phosphopeptides under basal conditions. Phosphorylation was increased approximately 3-fold in one of these peptides following forskolin treatment, and this change was eliminated by mutation of residue Ser605. Thus, phosphorylation of Ser605 is essential for cAMP-mediated inhibition of NHE3. In addition, Ser634 is also required for the effect of cAMP, even though this residue does not become phosphorylated upon activation of PKA. PMID- 9353336 TI - Human N-myristoyltransferase amino-terminal domain involved in targeting the enzyme to the ribosomal subcellular fraction. AB - N-Myristoyltransferase (NMT) catalyzes the cotranslational acylation with myristic acid of the NH2-terminal glycines of a number of cellular and viral proteins. Most of the in vitro NMT activity (60-85%) in isoosmotic cell homogenates of human lymphoblastic leukemia (i.e. CEM and MOLT-4) and cervical carcinoma (i.e. HeLa) cells was shown to be associated with the ribosomal subcellular fractions by differential centrifugation. Also found in the ribosomal fractions was a approximately 60-kDa protein that was specifically immunoblotted with an anti-human NMT (hNMT) peptide antibody. This approximately 60-kDa protein was stable in the presence of proteolytic enzyme inhibitors but was gradually converted into a approximately 46-kDa species when stored in the absence of protease inhibitors. Sucrose density gradient centrifugation of the ribosomal fraction resulted in the hNMT activity sedimenting exactly coincident with the 260 nm absorption profile and exhibiting A260/A280 absorption ratios >1.8, indicating an association of NMT with putative ribosomal particle(s)/subunit(s). The subcellular targeting of hNMT was also examined by immunoblotting subcellular fractions from HeLa cells transfected with plasmids containing FLAG epitope tagged hNMT inserts corresponding either to the originally assigned hNMT gene or to an alternative open reading frame initiated from an in-frame start site upstream from the assumed hNMT start site. Anti-FLAG immunoblotting of cells transfected with a plasmid containing the larger insert revealed FLAG-NMT primarily in the ribosomal fraction with an apparent molecular mass similar to the approximately 60-kDa native hNMT. In contrast, immunoblotting of cells transfected with a plasmid containing the smaller insert identified a approximately 50-kDa FLAG-NMT predominantly in the cytosolic fraction. An analysis of mixtures of CEM ribosomes and serial dilutions of purified recombinant FLAG-NMTs demonstrated that the approximately 60-kDa FLAG-NMT binds ribosomes with higher affinity than the approximately 50-kDa FLAG-NMT. These in vivo and in vitro subcellular targeting and recombinant expression experiments identify a native hNMT that is 10-12 kDa larger than the enzyme predicted by the originally assigned hNMT gene and which is apparently translated from an alternative up-stream start site. The data also indicate that although the unique NH2-terminal residues encoded by this larger open reading frame are not required for in vitro catalytic activity, they do provide signal(s) involved in targeting hNMT to the ribosomal subcellular fraction where cotranslational N-myristoylation occurs. PMID- 9353337 TI - Identification and characterization of Saccharomyces cerevisiae dihydrosphingosine-1-phosphate phosphatase. AB - We have identified the yeast sphingosine resistance gene (YSR2) of Saccharomyces cerevisiae as encoding a protein that specifically dephosphorylates dihydrosphingosine 1-phosphate (DHS-1-P), and we refer to this protein as dihydrosphingosine-1-phosphate phosphatase. Overexpression of YSR2 conferred sphingosine resistance to the dihydrosphingosine-1-P lyase-defective mutant (JS16) of S. cerevisiae, which is hypersensitive to sphingosine. The ysr2Delta deletion mutant of S. cerevisiae accumulated DHS-1-P compared with its wild type strain upon labeling with D-erythro-[4, 5-3H]dihydrosphingosine, whereas overexpression of YSR2 increased dephosphorylation of DHS-1-P. An epitope-tagged fusion protein (YSR2-Flag) was partially purified and found to specifically dephosphorylate DHS-1-P to yield dihydrosphingosine. YSR2 failed to dephosphorylate ceramide 1-phosphate or phosphatidic acid. Functionally, the mutant bearing the ysr2Delta deletion decreased labeling of sphingolipids and increased labeling of glycerolipids dramatically following in vivo labeling with D-erythro-[3H]dihydrosphingosine, but it slightly affected labeling of sphingolipids with inositol. Taken together, these results identify YSR2 as dihydrosphingosine-1-phosphate phosphatase. They also raise the intriguing possibility that phosphorylation followed by dephosphorylation is required for incorporation of exogenous long chain sphingoid bases into sphingolipids. PMID- 9353338 TI - Cloning and characterization of a human STE20-like protein kinase with unusual cofactor requirements. AB - We cloned and characterized a novel human member of the STE20 serine/threonine protein kinase family named mst-3. Based on its domain structure, mst-3 belongs to the SPS1 subgroup of STE20-like proteins, which includes germinal center (GC) kinase, hematopoietic progenitor kinase (HPK), kinase homologous to STE20/SPS-1 (KHS), kinases responsive to stress (KRS1/2), the mammalian STE20-like kinases (mst1/2), and the recently published STE20/oxidant stress response kinase SOK-1. mst-3 is most closely related to SOK-1, with 88% amino acid similarity in the kinase domain. The similarity of the mst-3 kinase domain to STE20 is 42%. The mst 3 transcript is ubiquitously expressed, and the protein was found in all human, mouse, and monkey cell lines tested. An in vitro kinase assay showed that mst-3 can phosphorylate basic exogenous substrates as well as itself. Interestingly, mst-3 prefers Mn2+ to Mg2+ as a divalent cation and can use both GTP and ATP as phosphate donors. Like SOK-1, mst-3 is activated by autophosphorylation. However, a physiological stimulus of mst-3 activity was not identified. mst-3 activity does not change upon exposure to several mitogenic and stress stimuli. Overexpression of mst-3 wild-type or kinase dead protein affects neither the extracellular signal-regulated kinases (ERK1/2 or ERK6), c-Jun N-terminal kinase (JNK), p38, nor pp70S6 kinase, suggesting that mst-3 is part of a novel signaling pathway. PMID- 9353339 TI - Requirement for protein kinase C theta for cell cycle progression and formation of actin stress fibers and filopodia in vascular endothelial cells. AB - Activation of the protein kinase C (PKC) family with phorbol esters induces endothelial proliferation and angiogenesis, but which of the events that constitute angiogenesis are affected by individual members of the PKC family is unknown. In rat capillary endothelial (RCE) cells, serum stimulation increased expression of a single PKC isoenzyme, PKCtheta, and its translocation to the periphery. Conditional overexpression of a dominant-negative mutant of PKCtheta markedly inhibited RCE proliferation, as well as closure of a "wound" by RCE migration and formation of capillary rings and tubules in vitro. PKCtheta inhibition delayed the endothelial cell cycle at the G2/M phase and prevented formation of actin stress fibers and filopodia but not lamellipodia. The defect in cell morphology and wound closure in PKCtheta-kn cells was reversed by overexpressing kinase-active PKCtheta, indicating that these RCE functions depend upon PKCtheta substrates. Thus, PKCtheta is required for multiple processes essential for angiogenesis and wound repair, including endothelial mitosis, maintenance of a normal actin cytoskeleton, and formation of an enclosed tube. PMID- 9353340 TI - Characterization of the phosphorylation sites involved in G protein-coupled receptor kinase- and protein kinase C-mediated desensitization of the alpha1B adrenergic receptor. AB - Catecholamines as well as phorbol esters can induce the phosphorylation and desensitization of the alpha1B-adrenergic receptor (alpha1BAR). In this study, phosphoamino acid analysis of the phosphorylated alpha1BAR revealed that both epinephrine- and phorbol ester-induced phosphorylation predominantly occurs at serine residues of the receptor. The findings obtained with receptor mutants in which portions of the C-tail were truncated or deleted indicated that a region of 21 amino acids (393-413) of the carboxyl terminus including seven serines contains the main phosphorylation sites involved in agonist- as well as phorbol ester-induced phosphorylation and desensitization of the alpha1BAR. To identify the serines invoved in agonist- versus phorbol ester-dependent regulation of the receptor, two different strategies were adopted, the seven serines were either substituted with alanine or reintroduced into a mutant lacking all of them. Our findings indicate that Ser394 and Ser400 were phosphorylated following phorbol ester-induced activation of protein kinase C, whereas Ser404, Ser408, and Ser410 were phosphorylated upon stimulation of the alpha1BAR with epinephrine. The observation that overexpression of G protein-coupled kinase 2 (GRK2) could increase agonist-induced phosphorylation of Ser404, Ser408, and Ser410, strongly suggests that these serines are the phosphorylation sites of the alpha1BAR for kinases of the GRK family. Phorbol ester-induced phosphorylation of the Ser394 and Ser400 as well as GRK2-mediated phosphorylation of the Ser404, Ser408, and Ser410, resulted in the desensitization of alpha1BAR-mediated inositol phosphate response. This study provides generalities about the biochemical mechanisms underlying homologous and heterologous desensitization of G protein-coupled receptors linked to the activation of phospholipase C. PMID- 9353341 TI - Alternatively spliced focal adhesion kinase in rat brain with increased autophosphorylation activity. AB - pp125 focal adhesion kinase (FAK), a cytoplasmic tyrosine kinase transducing signals initiated by integrin engagement and G protein-coupled receptors, is highly expressed in brain. FAK from brain had a higher molecular weight and an increased autophosphorylation activity, than from other tissues. In addition to a 9-base insertion in the 3'-coding region, which defines FAK+, rat striatal FAK mRNAs contained several additional short exons, coding for peptides of 28, 6, and 7 residues, respectively (termed boxes 28, 6, and 7), surrounding the autophosphorylated Tyr-397. In transfected COS 7 cells, the presence of boxes 6 and 7 conferred an increased overall tyrosine phosphorylation, a higher phosphorylation of Tyr-397 assessed with a phosphorylation state-specific antibody, and a more active autophosphorylation in immune precipitates. The presence of box 28 did not alter further these parameters. Two-dimensional phosphopeptide maps of hippocampal FAK were identical to those of FAK+6,7. The presence of the various exons did not alter the interaction of FAK with c-Src, n Src, or Fyn. Thus, several splice isoforms of FAK are preferentially expressed in rat brain, some of which have an increased autophosphorylation activity, suggesting that FAK may have specific properties in neurons. PMID- 9353342 TI - Regulation of human chemokine receptors CXCR4. Role of phosphorylation in desensitization and internalization. AB - Members of the chemokine receptor family CCR5 and CXCR4 have recently been shown to be involved in the entry of human immunodeficiency virus (HIV) into target cells. Here, we investigated the regulation of CXCR4 in rat basophilic leukemia cells (RBL-2H3) stably transfected with wild type (Wt CXCR4) or a cytoplasmic tail deletion mutant (DeltaCyto CXCR4) of CXCR4. The ligand, stromal cell derived factor-1 (SDF-1) stimulated higher G-protein activation, inositol phosphate generation, and a more sustained calcium elevation in cells expressing DeltaCyto CXCR4 relative to Wt CXCR4. SDF-1 and phorbol 12-myristate 13-acetate (PMA), but not a membrane permeable cAMP analog induced rapid phosphorylation as well as desensitization of Wt CXCR4. Phosphorylation of DeltaCyto CXCR4 was not detected under any of these conditions. Despite lack of receptor phosphorylation, calcium mobilization by SDF-1 in DeltaCyto CXCR4 cells was partially desensitized by prior treatment with SDF-1. Of interest, the rapid release of calcium was inhibited without affecting the sustained calcium elevation, indicating independent regulatory pathways for these processes. PMA completely inhibited phosphoinositide hydrolysis and calcium mobilization in Wt CXCR4 but only partially inhibited these responses in DeltaCyto CXCR4. cAMP also partially inhibited these responses in both Wt CXCR4 and DeltaCyto CXCR4. SDF-1, PMA, and cAMP caused phosphorylation of phospholipase Cbeta3 in Wt and DeltaCyto CXCR4 cells. Both SDF-1 as well as PMA induced rapid internalization of Wt CXCR4. SDF-1 but not PMA induced internalization of DeltaCyto CXCR4 albeit at reduced levels relative to Wt CXCR4. These results indicate that signaling and internalization of CXCR4 are regulated by receptor phosphorylation dependent and independent mechanisms. Desensitization of CXCR4 signaling, independent of receptor phosphorylation, appears to be a consequence of the phosphorylation of phospholipase Cbeta3. PMID- 9353343 TI - Modulation of AUUUA response element binding by heterogeneous nuclear ribonucleoprotein A1 in human T lymphocytes. The roles of cytoplasmic location, transcription, and phosphorylation. AB - The heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) shuttles between the cytoplasm and nucleus and plays important roles in RNA metabolism. Whereas nuclear hnRNP A1 has been shown to bind intronic sequences and modulate splicing, cytoplasmic hnRNP A1 is associated with poly(A)+ RNA, indicating different RNA ligand specificity. Previous studies indicated that cytoplasmic hnRNP A1 is capable of high-affinity binding of reiterated AUUUA sequences (ARE) that have been shown to modulate mRNA turnover and translation. Through a combination of two-dimensional gel and proteolysis studies, we establish hnRNP A1 (or structurally related proteins that are post-translationally regulated in an identical manner) as the dominant cytoplasmic protein in human T lymphocytes capable of interacting with the ARE contained within the context of full-length granulocyte-macrophage colony-stimulating factor mRNA. We additionally demonstrate that cytoplasmic hnRNP A1 preferentially binds ARE relative to pre mRNAs in both cross-linking and mobility shift experiments. RNA polymerase II inhibition increased the binding of ARE (AUBP activity) and poly(U)-Sepharose by cytoplasmic hnRNP A1, while nuclear hnRNP A1 binding was unaffected. Nuclear and cytoplasmic hnRNP A1 could be distinguished by the differential sensitivity of their RNA binding to diamide and N-ethylmaleimide. The increase in AUBP activity of cytoplasmic hnRNP A1 following RNA polymerase II inhibition correlated with serine-threonine dephosphorylation, as determined by inhibitor and metabolic labeling studies. Thus, cytoplasmic and nuclear hnRNP A1 exhibit different RNA binding profiles, perhaps transduced through serine-threonine phosphorylation. These findings are relevant to the specific ability of hnRNP A1 to serve distinct roles in post-transcriptional regulation of gene expression in both the nucleus and cytoplasm. PMID- 9353344 TI - Organization of the neurofascin gene and analysis of developmentally regulated alternative splicing. AB - Neurofascin is an axonal member of the L1 subgroup of the immunoglobulin superfamily implicated in neurite extension in the course of embryonic development. Here we have isolated and characterized the gene encoding chicken neurofascin. Comparison of genomic sequences with cDNA sequences provides the structure and localization of intron/exon boundaries and indicates that neurofascin isoforms are generated by alternative splicing of its pre-mRNA. The neurofascin gene is composed of 33 exons distributed over 72 kilobases. Each of the six immunoglobulin- and five fibronectin-type III-like domains is encoded by two exons. While introns between domains are of phase 1, others are of phase 0, 1, or 2. Alternative splicing of neurofascin is developmentally regulated as shown by polymerase chain reaction analysis. Furthermore, plasmid libraries from long range polymerase chain reaction-amplified cDNA of neurofascin were used to examine and quantify the distribution of alternatively spliced exons in individual neurofascin molecules. We found 50 different neurofascin isoforms at different developmental stages and revealed the existence of one major "early" in comparison with multiple "late" neurofascin isoforms. PMID- 9353345 TI - Engagement of P-selectin glycoprotein ligand-1 enhances tyrosine phosphorylation and activates mitogen-activated protein kinases in human neutrophils. AB - During inflammation, P-selectin on activated platelets and endothelial cells initiates adhesion of leukocytes through interactions with P-selectin glycoprotein ligand-1 (PSGL-1). We investigated whether ligation of PSGL-1 also transmits signals into leukocytes. Neutrophils incubated with anti-PSGL-1 monoclonal antibodies, but not with Fab fragments of these antibodies, rapidly increased tyrosine phosphorylation of proteins with relative molecular masses of 105-120, 70-84, and 42-44 kDa. PSGL-1-dependent adhesion of neutrophils to P selectin increased tyrosine phosphorylation of similarly sized proteins. Cytochalasin B did not prevent the tyrosine phosphorylation induced by ligation of PSGL-1, suggesting that an intact cytoskeleton is not required for signaling. Engagement of PSGL-1 activated the GTPase Ras through a mechanism that did not require tyrosine phosphorylation of PSGL-1 or association of the Shc.Grb2.Sos1 complex with PSGL-1. Engagement of PSGL-1 activated the 42-44-kDa extracellular signal-regulated kinase family of mitogen-activated protein (MAP) kinases through a pathway that required activation of the MAP kinase kinase. Ligation of PSGL-1 also stimulated secretion of interleukin-8. The tyrosine kinase inhibitor, genistein, blocked tyrosine phosphorylation and secretion of interleukin-8, whereas the MAP kinase kinase inhibitor PD98059 partially inhibited secretion of interleukin-8. Tyrosine phosphorylation stimulated through PSGL-1 on selectin tethered leukocytes may propagate a signaling cascade that is integrated with signals generated by other mediators. PMID- 9353346 TI - Requirement of integrin beta3 tyrosine 747 for beta3 tyrosine phosphorylation and regulation of alphavbeta3 avidity. AB - Leukocytes and platelets require stimulation for optimal beta3 integrin receptor function, whereas beta3 function is constitutive in many other cells. The molecular mechanisms that enhance integrin function in stimulated hematopoietic cells are poorly understood. Phosphorylation of the beta3 cytoplasmic tail is a recently described but prevalent phenomenon, with unknown effects on alphavbeta3 function. Here, we show that mutation of the beta3 cytoplasmic tail tyrosine 747 to phenylalanine (Y747F) prevents beta3 tyrosine phosphorylation in two cell lines. Whereas this mutation has no effect on alphavbeta3-mediated adhesion in a cell with constitutive beta3 function, it completely abolishes adhesion and clot retraction by a cell that requires stimulation for beta3 function. Ligand-induced conformational change as detected by LIBS-1 antibody occurs normally in Y747F mutant alphavbeta3. Thus, tyrosine 747 of beta3 is required for stimulation of alphavbeta3-mediated adhesion, probably due to its phosphorylation. Because the motif in beta3 required for tyrosine phosphorylation is shared by several integrin beta-chains, this may be a conserved mechanism for regulation of integrin-dependent adhesion. PMID- 9353347 TI - Selective inhibition of mitogen-induced transactivation of the HIV long terminal repeat by carboxyamidotriazole. Calcium influx blockade represses HIV-1 transcriptional activation. AB - Carboxyamidotriazole (CAI) is a calcium influx inhibitor that has both antiproliferative and antimetastatic activities. Pretreatment of human T-cells with micromolar concentrations of CAI causes a near complete inhibition of calcium-regulated mitogen-induced transcription from the human immunodeficiency virus (HIV) long terminal repeat (LTR). This inhibition is selective since other mitogen-activated gene regulatory elements, such as the 12-O-tetradecanoylphorbol 13-acetate response element, are not influenced by the drug. HIV LTR transcription inhibition is maximal at 1.0 microM CAI, requires a pretreatment interval of at least 8 h for optimum inhibition, and shows no acute interference with the growth properties of the cells. Moreover, the inhibition is rapidly reversible upon removal of the drug from the medium. Studies to identify enhancer elements within the HIV LTR that are functionally sensitive to low-dose long-term pretreatment with CAI indicate that the NF-kappaB-binding sites are among the major targets of drug action. In vitro DNA binding studies with nuclear extracts prepared from mitogen-induced T-cells stimulated in the presence of CAI indicate that the drug differentially influences the calcium-regulated downstream signal transduction pathways necessary for specific NF-kappaB DNA binding activity at the two kappaB sites within the HIV LTR. Studies with ionomycin and thapsigargin show that repression is specific for selected modes of inducible calcium entry and indicate that, in T-cells, a major mechanism of CAI action is to modulate calcium influx at a level that is proximal to the regulated release of calcium from intracellular stores. Measurement of calcium influx in CAI-treated cells reveals a dramatic and reversible inhibition of mitogen-induced calcium influx. These results indicate that CAI can be an important and effective pharmacological tool for analysis of the calcium-dependent modulation of HIV LTR transcription. PMID- 9353348 TI - Dystrophin-glycoprotein complex is monomeric and stabilizes actin filaments in vitro through a lateral association. AB - The native molecular weight of the dystrophin-glycoprotein complex and its effect on actin depolymerization and polymerization were examined. First, we determined that the native molecular weight of purified dystrophin-glycoprotein complex is only large enough (Mr 1,200,000) to contain one copy of each protein in the complex, including dystrophin. Using different approaches, we also demonstrated that dystrophin-glycoprotein complex significantly protected a fraction of actin filaments from disassembly, while individual recombinant actin binding fragments of dystrophin or calpain-digested dystrophin-glycoprotein complex had no effect on F-actin depolymerization. The protective effect of dystrophin-glycoprotein complex on F-actin depolymerization saturated at a dystrophin:actin molar ratio of 0.04, corresponding to 1 dystrophin/25 actin monomers, which is highly consistent with the 1:24 stoichiometry of dystrophin-glycoprotein complex binding to F-actin previously measured at equilibrium. However, dystrophin-glycoprotein complex did not bind G-actin or alter the kinetics or extent of actin polymerization. This excluded the possibility that dystrophin-glycoprotein complex inhibited actin depolymerization by capping the ends of actin filaments. It therefore appears that actin binding domains separated on the dystrophin molecule from each other by almost 1,200 amino acids act in concert to protect F actin from depolymerization. Our data suggest that dystrophin stabilizes F-actin in vitro by binding alongside an actin filament and bridging actin monomers in a manner analogous to the actin side binding protein tropomyosin. It is noteworthy that we did not find any effect of skeletal muscle tropomyosin on dystrophin glycoprotein complex binding to F-actin. This indicates that dystrophin glycoprotein complex and tropomyosin may simultaneously bind the same actin filament and identifies another feature that distinguishes dystrophin from the other proteins in the actin-cross-linking superfamily. PMID- 9353349 TI - Interferon-resistant human melanoma cells are deficient in ISGF3 components, STAT1, STAT2, and p48-ISGF3gamma. AB - The mechanism of IFN resistance was examined in three long-term cell lines, SK MEL-28, SK-MEL-3, and MM96, exhibiting significant variation in responsiveness to the antiproliferative and antiviral effects of type I IFNs. The JAK-STAT components involved in IFN signal transduction were analyzed in detail. After exposure to IFN, activation of the IFN type I receptor-linked tyrosine kinases, JAK-1 and TYK-2, was detected at similar levels in both IFN-sensitive and IFN resistant cell types, indicating that IFN resistance did not result from a deficiency in signaling at the level of receptor-associated kinase activation. However, analysis of ISGF3 transcription factor components, STAT1, STAT2, and p48 ISGF3gamma, revealed that their expression and activation correlated with cellular IFN responsiveness. The analysis was extended to also include IFN sensitive primary melanocytes, three additional IFN-resistant melanoma cell lines, and seven cell cultures recently established from melanoma patient biopsies. It was consistently observed that the most marked difference in ISGF3 was a lack of STAT1 in the resistant versus the sensitive cells. Transfection of the IFN-resistant MM96 cell line to express increased levels of STAT1 protein partially restored IFN responsiveness in an antiviral assay. We conclude that a defect in the level of STAT1 and possibly all three ISGF3 components in IFN resistant human melanoma cells may be a general phenomenon responsible for reduced cellular responsiveness of melanomas to IFNs. PMID- 9353350 TI - The binding of T cell-expressed P-selectin glycoprotein ligand-1 to E- and P selectin is differentially regulated. AB - The HECA452 carbohydrate epitope, also termed cutaneous lymphocyte antigen, is known to bind to E-selectin and defines a human T cell subset preferentially found in inflamed skin. Activated T cells can express a functional form of the P selectin glycoprotein ligand-1 (PSGL-1), the major ligand known for P-selectin. Here we show that PSGL-1 can exist in two forms, of which only one carries the HECA452 epitope and binds to E-selectin, while the other only binds to P selectin. We have analyzed the glycoprotein ligands for E- and P-selectin on the mouse CD8+ T cell clone 4G3 at 4, 8, and 12 days after antigen-specific activation. Only at day 4 did the cells bind to E-selectin, whereas cells at all three activation stages bound to P-selectin. Expression of the HECA452 epitope correlated with E-selectin binding. In affinity isolation experiments, PSGL-1 was isolated as the major ligand by E-selectin-IgG and by P-selectin-IgG; however, PSGL-1 only bound to E-selectin at day 4, whereas it bound to P-selectin at all three activation stages. Immunoprecipitated PSGL-1 from cells at day 4, but not from cells at days 8 and 12, was recognized in immunoblots by monoclonal antibody HECA452. In immunoblots of total extracts of cells at day 4, HECA452 recognized a 240/140-kDa pair of protein bands as the major antigen. These bands could be completely removed by depletion of cell extracts with anti-PSGL-1 antibodies. Our data suggest that the carbohydrate requirements for binding of PSGL-1 to P selectin differ from those necessary for binding to E-selectin. Furthermore, we conclude that the major glycoprotein carrier for the HECA452 epitope on activated 4G3 cells is PSGL-1. PMID- 9353351 TI - Both the catalytic and regulatory domains of protein kinase C chimeras modulate the proliferative properties of NIH 3T3 cells. AB - Protein kinase C (PKC) isozymes exhibit important differences in terms of their regulation and biological functions. Not only may some PKC isoforms be active and others not for a given response, but the actions of different isoforms may even be antagonistic. In NIH 3T3 cells, for example, PKCdelta arrests cell growth whereas PKCepsilon stimulates it. To probe the contribution of the regulatory and the catalytic domains of PKC isozymes to isozyme-specific responses, we prepared chimeras between the regulatory and the catalytic domains of PKCalpha, -delta, and -epsilon. These chimeras, which preserve the overall structure of the native PKC enzymes, were stably expressed in mouse fibroblasts. A major objective was to characterize the growth properties of the cells that overexpress the various PKC constructs. Our data demonstrate that both the regulatory and the catalytic domains play roles in cell proliferation. The regulatory domain of PKCepsilon enhanced cell growth in the absence or presence of phorbol 12-myristate 13 acetate (PMA), and, in the presence of PMA, all chimeras with the PKCepsilon regulatory domain also gave rise to colonies in soft agar; the role of the catalytic domain of PKCepsilon was evident in the PMA-treated cells that overexpressed the PKC chimera containing the delta regulatory and the epsilon catalytic domains (PKCdelta/epsilon). The important contribution of the PKCepsilon catalytic domain to the growth of PKCdelta/epsilon-expressing cells was also evident in terms of a significantly increased saturation density in the presence of PMA, their formation of foci upon PMA treatment, and the induction of anchorage-independent growth. Aside from the growth-promoting effect of PKCepsilon, we have shown that most chimeras with PKCalpha and -delta regulatory domains inhibit cell growth. These results underscore the complex contributions of the regulatory and catalytic domains to the overall behavior of PKC. PMID- 9353352 TI - Functional properties of replication fork assemblies established by the bacteriophage lambda O and P replication proteins. AB - We have used a set of bacteriophage lambda and Escherichia coli replication proteins to establish rolling circle DNA replication in vitro to permit characterization of the functional properties of lambda replication forks. We demonstrate that the lambda replication fork assembly synthesizes leading strand DNA chains at a physiological rate of 650-750 nucleotides/s at 30 degrees C. This rate is identical to the fork movement rate we obtained using a minimal protein system, composed solely of E. coli DnaB helicase and DNA polymerase III holoenzyme. Our data are consistent with the conclusion that these two key bacterial replication proteins constitute the basic functional unit of a lambda replication fork. A comparison of rolling circle DNA replication in the minimal and lambda replication systems indicated that DNA synthesis proceeded for more extensive periods in the lambda system and produced longer DNA chains, which averaged nearly 200 kilobases in length. The higher potency of the lambda replication system is believed to result from its capacity to mediate efficient reloading of DnaB helicase onto rolling circle replication products, thereby permitting reinitiation of DNA chain elongation following spontaneous termination events. E. coli single-stranded DNA-binding protein and primase individually stimulated rolling circle DNA replication, but they apparently act indirectly by blocking accumulation of inhibitory free single-stranded DNA product. Finally, in the course of this work, we discovered that E. coli DNA polymerase III holoenzyme is itself capable of carrying out significant strand displacement DNA synthesis at about 50 nucleotides/s when it is supplemented with E. coli single-stranded DNA-binding protein. PMID- 9353353 TI - Evidence for endothelin involvement in the pulmonary vasoconstrictor response to systemic hypoxia in the isolated rat lung. AB - We investigated the effect of systemic hypoxia (Krebs-Henseleit solution gassed with 5% CO2/95% N2) on an isolated, perfused rat lung. Hypoxia resulted in a slowly developing sustained increase in pulmonary perfusion pressure (PPP) accompanied by an increase in lung weight (LW). The endothelin (ET) receptor antagonists BQ123 (3 and 10 microM), BQ788 (3 microM) and bosentan (1.5 and 5 microM) all attenuated the hypoxia-induced increases in LW and PPP. In addition, phosphoramidon (1 microM), an ET-converting enzyme inhibitor, also significantly attenuated the hypoxia-induced increases in PPP and LW. The use of two agents that alter peptide secretion, phalloidin (10 and 50 nM) and colchicine (100 nM), and the peptide synthesis inhibitor cycloheximide (5 microM) all significantly attenuated the hypoxia-induced increases in PPP and LW. The increase in PPP and LW after the onset of hypoxia was accompanied by an increase in perfusate levels of ET-1 compared with normoxic time-matched controls. The results show that in this model, systemic hypoxia is capable of causing a sustained vasoconstriction and increased LW. The fact that these increases can be attenuated by an ET converting enzyme inhibitor, ET receptor antagonists and agents that block peptide synthesis and secretion, together with the increase in perfusate levels of ET-1, suggests that ET production and release contribute to the changes seen. PMID- 9353354 TI - Atrial natriuretic peptide inhibits evoked catecholamine release by altering sensitivity to calcium. AB - Natriuretic peptides are cyclized peptides produced by cardiovascular and neural tissues. These peptides inhibit various secretory responses such as the release of renin, aldosterone and autonomic neurotransmitters. This report tests the hypothesis that atrial natriuretic peptide reduces dopamine efflux from an adrenergic cell line, rat pheochromocytoma cells, by suppressing intracellular calcium concentrations. The L-type calcium channel inhibitor, nifedipine, markedly suppressed dopamine release from depolarized PC12 cells, suggesting that calcium entering through this channel was the predominant stimulus for dopamine efflux. Atrial natriuretic peptide maximally reduced depolarization-evoked dopamine release 20 +/- 3% at a concentration of 100 nM and this effect was abolished by nifedipine, but not by pretreatment with the N-type calcium channel inhibitor, omega-conotoxin, or an inhibitor of calcium-induced calcium release, ryanodine. In cells loaded with Fura-2, atrial natriuretic peptide both augmented depolarization-induced increases of intracellular free calcium concentrations and accelerated the depolarization-induced quenching of the Fura-2 signal by manganese, findings consistent with enhanced conductivity of calcium channels. Dopamine efflux induced by either the calcium ionophore, A23187, or staphylococcal alpha toxin was attenuated by atrial natriuretic peptide. Additionally, a natriuretic peptide interacting solely with the natriuretic peptide C receptor in these cells, C-type natriuretic peptide, also suppressed calcium-induced dopamine efflux in permeabilized cells. These data are consistent with natriuretic peptides attenuating catecholamine exocytosis in response to calcium but inconsistent with the neuromodulatory effect resulting from a reduction in intracellular calcium concentrations within pheochromocytoma cells. PMID- 9353355 TI - Different contributions of cytochrome P450 2C19 and 3A4 in the oxidation of omeprazole by human liver microsomes: effects of contents of these two forms in individual human samples. AB - Omeprazole 5-hydroxylation and sulfoxidation activities were determined in liver microsomes of different humans whose levels of individual forms of cytochrome P450 (P450 or CYP) varied. Correlation coefficients between omeprazole 5 hydroxylation activities (when determined at a substrate concentration of 10 microM) and S-mephenytoin 4'-hydroxylation and testosterone 6beta-hydroxylation activities were found to be 0.64 and 0.67, respectively, in liver microsomes of 84 human samples examined. Omeprazole sulfoxidation activities in these human samples were correlated with testosterone 6beta-hydroxylation activities (r = 0. 86). Omeprazole 5-hydroxylation by liver microsomes of a human sample that contained relatively high levels of CYP3A4 and low levels of CYP2C19 were inhibited very significantly by ketoconazole and anti-CYP3A4 antibodies, although a human sample having high in CYP2C19 and low in CYP3A4 was found to be sensitive toward fluvoxamine and anti-CYP2C9 antibodies. Sulfaphenazole (at 100 microM) did not affect the omeprazole 5-hydroxylation and sulfoxidation catalyzed by human liver microsomes. Both recombinant human CYP2C19 and CYP3A4 enzymes had activities for omeprazole 5-hydroxylation, with low Km and high Vmax values for the former enzyme and high Km and low Vmax values for the CYP3A4. These results suggest that contributions of CYP2C19 and CYP3A4 in the omeprazole 5 hydroxylation depend upon the ratio of these two P450 levels in human liver microsomes. Omeprazole 5-hydroxylation activities of different human samples were found to be related to predicted values calculated from the kinetic parameters of recombinant enzymes and the levels of liver microsomal CYP2C19 and CYP3A4 enzymes. Finally, when recombinant human CYP2C19 and CYP3A4 were mixed at levels found in different human samples, relatively similar profiles of omeprazole oxidation by the recombinant and microsomal enzyme systems were determined by analysis of high-performance liquid chromatography. These results suggest that both CYP2C19 and CYP3A4 are involved in the 5-oxidation of omeprazole (at a substrate concentration of 10 microM) in human liver microsomes and that contributions of these P450 enzymes depend on the compositions of CYP2C19 and CYP3A4 in liver. PMID- 9353356 TI - Mechanisms for the paradoxical resistance to d-tubocurarine during immobilization induced muscle atrophy. AB - This study investigated whether immobilization-induced hyposensitivity to d tubocurarine (dTC), up-regulation of acetylcholine receptors (AChRs) and changes in fiber size and motor endplate size persist indefinitely and whether they are causally related. Unilateral disuse of the tibialis muscle was produced in adult rats by pinning the knee and ankle joints at 90 degrees flexion. The contralateral unpinned and a separate group of sham-pinned legs served as controls. After 7, 14 or 28 days of disuse, the in vivo dose of dTC that produced 50% depression of nerve-evoked twitch (ED50) in the tibialis muscle increased 3.0 , 3. 2- and 2.1-fold (P < .05), and membrane AChRs increased 6.0- (P < . 05), 6.3 (P > .05) and 1.2-fold (P > .395) relative to control, respectively. Disuse caused muscle fiber atrophy (P < .01) but did not affect endplate size. Hence, the ratio of endplate size to fiber size increased. There was a transient increase in gene expression of all (including de novo expression of the gamma) subunits of the AChR, peaking at day 7 and returning to normal by day 28 of immobilization. The ED50 of dTC correlated directly with AChRs (R2 = 0.51; P < .0001) or the ratio of endplate size to fiber size (R2 = 0. 30; P < .001), and inversely with fiber size (R2 = 0.43, P < .0001). It is proposed that acting together, but not singly, the changes in AChRs, fiber size and relative endplate size contribute to the magnitude and time course of the resistance to dTC produced by chronic disuse. PMID- 9353357 TI - Beta-2 adrenergic activation of L-type Ca++ current in cardiac myocytes. AB - The whole-cell patch-clamp and intracellular perfusion techniques were used for studying the effects of a beta-2 adrenergic receptor activation on the L-type Ca current (ICa) in frog ventricular myocytes. The beta-2 adrenergic agonist zinterol increased ICa in a concentration-dependent manner with an EC50 (i.e., the concentration of zinterol at which the response was 50% of the maximum) of 2.2 nM. The effect of zinterol was essentially independent of the membrane potential. The stimulatory effect of zinterol was competitively antagonized by ICI 118,551, a beta-2 adrenergic antagonist. The maximal stimulatory effect of zinterol was comparable in amplitude to the effect of a saturating concentration (1 or 10 microM) of isoprenaline, a nonselective beta adrenergic agonist. Moreover, 3-isobutyl-1-methylxanthine (100 microM), a nonselective phosphodiesterase inhibitor, or forskolin (10 microM), a direct activator of adenylyl cyclase, had no additive effects in the presence of 0.1 microM zinterol. Zinterol had a long lasting action on frog ICa because after washout of the drug, ICa returned to basal level with a time constant of 17 min. An application of acetylcholine (1 microM) during this recovery phase promptly reduced ICa back to its basal level suggesting a persistent activation of adenylyl cyclase due to a slow dissociation rate constant of zinterol from its receptor. Zinterol also increased ICa in rat ventricular and human atrial myocytes, and the maximal effect was obtained at 10 and 1 microM, respectively. In all three preparations, intracellular perfusion with 20 microM PKI(15-22), a highly selective peptide inhibitor of cAMP-dependent protein kinase, completely antagonized the stimulatory effect of zinterol on ICa. We conclude that beta-2 adrenergic receptor activation produces a strong increase in ICa in frog, rat and human cardiac myocytes which is due to stimulation of adenylyl cyclase and activation of cAMP-dependent phosphorylation. PMID- 9353358 TI - Prediction of species differences (rats, dogs, humans) in the in vivo metabolic clearance of YM796 by the liver from in vitro data. AB - The bioavailability after oral administration of (S)-(-)-2,8-dimethyl-3-methylene 1-oxa-8-azaspiro [4,5] decane-L-tartarate monohydrate (YM796), which is being developed as an antidementia drug, at a dose of 1 mg/kg was very low (3.4%) in rats, but considerably higher (16.1%) in dogs. The oral clearances (CLoral, Dose/AUCoral) in rats and dogs were, respectively, 300 and 18 times more than that already reported in humans. We have previously reported successful attempts to predict the in vivo hepatic metabolic clearance of YM796 from in vitro data in humans. In our study, the in vitro metabolism of YM796 was determined using liver microsomes prepared from both rats and dogs and we also investigated if the species difference observed in vivo could be quantitatively reproduced in vitro. In rats, total metabolite formation could be described by single component kinetics with a Km of 13.4 microM and a Vmax of 520 nmol/min/g liver. However, in dogs, total metabolite formation could be described by three components, as also reported for humans. The Km and Vmax values for the high-affinity, low-capacity component (Km1 and Vmax1) in dogs and humans were, respectively, 8.1 and 1.7 microM, and 10.9 and 1.2 nmol/min/g liver. The overall intrinsic metabolic clearances estimated from the in vitro studies (CLint,in vitro) for rats and dogs were 38.8 and 2.6 ml/min/g liver, respectively, being approximately 40 and 3 times more than that previously reported for humans (0.94 ml/min/g liver). The overall intrinsic hepatic clearances (CLint,in vivo) calculated from in vivo CLoral were 30.4, 3.4 and 0.73 ml/min/g liver for rats, dogs and humans, respectively, indicating that the in vivo hepatic clearance of YM796 can be predicted from in vitro metabolism data in each species. Thus, the pronounced species difference in the metabolic clearance observed in vivo can be quantitatively predicted from in vitro metabolic data using liver microsomes, and was predominantly due to the large difference in the Vmax values. PMID- 9353359 TI - An orally active selenium-based antihypertensive agent with restricted CNS permeability. AB - We report here the first orally active, selenium-based antihypertensive agent, and we demonstrate its restricted CNS permeability using inductively coupled plasma/mass spectroscopy (ICP/MS) and operant behavioral analysis. The biochemistry and pharmacology of selenium are subjects of intense current interest. As a consequence of the redox chemistry of the selenium moiety, phenylaminoalkyl selenides possess the remarkable characteristic of propagating a cycle of turnover-dependent local depletion of reduced ascorbate when processed by the key enzyme of catecholamine metabolism, dopamine-beta-monooxygenase. ICP/MS analysis was used to determine the pharmacokinetic parameters for selenide compounds after i.v. administration to anesthetized rats. Analysis of the data using a two-compartment pharmacokinetic model established very rapid initial clearance and a short beta-elimination half-life from blood. We developed an oxidative procedure for digestion and processing of tissue samples in order to obtain ICP/MS data on the tissue distributions of Se-containing metabolites after the administration of selenide compounds. The results establish that aromatic ring hydroxylation of the selenides results in a marked reduction in brain levels of Se-containing metabolites. The comparative effects of selenide compounds on locomotor activity and operant behavior were then investigated, and the results fully corroborate the ICP/MS analytical results. The novel compound, 4-hydroxy alpha-methyl-phenyl-2-aminoethyl selenide, exhibits both restricted CNS permeability and oral antihypertensive activity in spontaneously hypertensive rats. This compound is the first orally active selenium-based antihypertensive agent ever reported, and it possesses properties that are highly desirable in pharmacological agents being developed for treatment of chronic diseases such as hypertension. PMID- 9353360 TI - Pituitary adenylate cyclase-activating polypeptide-27 causes a biphasic chronotropic effect and atrial fibrillation in autonomically decentralized, anesthetized dogs. AB - We investigated the effects of a neuropeptide, pituitary adenylate cyclase activating polypeptide- (PACAP) 27, on the sinoatrial nodal pacemaker activity and the mechanisms for the cardiac effects of PACAP-27 in the autonomically decentralized heart of the anesthetized dog. PACAP-27 (0.01-0.3 nmol) injected into the sinus node artery increased followed by decreased sinus rate. PACAP-27 (0.1 and 0.3 nmol) caused atrial fibrillation spontaneously. After atropine, PACAP-27 never decreased but only increased sinus rate as did vasoactive intestinal peptide. However, propranolol did not affect the negative and positive chronotropic effects. Tetrodotoxin but not hexamethonium abolished the negative chronotropic response to PACAP-27 in atropine nontreated dogs, and tetrodotoxin also inhibited the positive chronotropic response by 34% in atropine-treated dogs. In atropine- and propranolol-treated dogs, positive chronotropic responses to PACAP-27 were inhibited by PACAP-(6-27), a PACAP receptor antagonist but not by vasoactive intestinal peptide (10-28), a vasoactive intestinal peptide receptor antagonist. These results indicate that PACAP-27 causes the negative chronotropic effect through the postganglionic parasympathetic nerve activation and it produces the positive chronotropic effect mediated by PACAP receptors with an activation of non-adrenergic, nonvasoactive intestinal peptide-ergic nerves at least in part in the dog heart. Atropine and tetrodotoxin abolished atrial fibrillation induced by PACAP-27 but other blockers did not. These results suggest that neurally released acetylcholine induced by PACAP-27 participates in the induction of atrial fibrillation. PMID- 9353361 TI - [3H]RY 80: A high-affinity, selective ligand for gamma-aminobutyric acidA receptors containing alpha-5 subunits. AB - The radiochemical synthesis and pharmacological properties are described of [3H]RY 80 (ethyl-8-acetylene-5, 6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5a][1, 4]benzodiazepine-3-carboxylate, [ethyl-3H]). This compound is one of a series of 8-substituted imidazobenzodiazepines that exhibits both high affinity and selectivity for gamma-aminobutyric acid (GABA)A receptors containing alpha-5 subunits. Saturable, high-affinity (Kd approximately 0.7 nM) binding of [3H]RY 80 was observed in hippocampal membranes. The maximum number (Bmax) of [3H]RY 80 binding sites was approximately 18% of that obtained with [3H]flunitrazepam, a radioligand that labels all "diazepam-sensitive" GABAA receptors. This value is consistent with previous estimates (10-20%) of the proportion of rat hippocampal GABAA receptors containing alpha-5 subunits determined by immunoprecipitation with selective antibodies and competition experiments using an alpha-5-selective ligand. In recombinant GABAA receptors composed of alpha-5 beta-3 gamma-2 subunits, the Kd of [3H]RY 80 (approximately 0.5 nM) was consistent with the value obtained in hippocampus, whereas the Bmax value was not significantly different from that obtained with [3H]flunitrazepam. The potencies of several benzodiazepine site ligands to inhibit [3H]RY 80 binding to hippocampal membranes were in agreement with the values obtained in recombinant (alpha-5 beta-3 gamma 2) GABAA receptors. [3H]RY 80 was used both in a "GABA shift" assay to correctly predict the in vivo actions of a novel, alpha-5-selective ligand and to characterize a population of GABAA receptors containing alpha-5 subunits in neonatal rat cortex. These findings demonstrate that [3H]RY 80 can be used as a radioligand to examine the properties of GABAA receptors containing alpha-5 subunits. PMID- 9353362 TI - Activation of histamine H3 receptors inhibits carrier-mediated norepinephrine release in a human model of protracted myocardial ischemia. AB - During protracted myocardial ischemia, ATP depletion promotes Na+ accumulation in sympathetic terminals and prevents vesicular storage of norepinephrine (NE). This forces the reversal of the neuronal uptake1 transporter, and NE is massively released (carrier-mediated release). We had shown that histamine H3 receptors (H3Rs) modulate ischemic NE release in animals. We have now used a human model of protracted myocardial ischemia to investigate whether H3Rs may control carrier mediated NE release. Surgical specimens of human atrium were incubated in anoxic conditions. NE release increased approximately 7-fold within 70 min of anoxia. This release was carrier mediated because it was Ca++ independent and inhibited by the uptake1 inhibitor desipramine. Furthermore, the Na+/H+ exchanger (NHE) inhibitors ethyl-isopropyl-amiloride and HOE 642, and the Na+ channel blocker tetrodotoxin inhibited NE release, whereas the Na+ channel activator aconitine potentiated it. The selective H3R agonist imetit decreased NE release, an effect that was blocked by each of the H3R antagonists thioperamide and clobenpropit. Notably, imetit acted synergistically with ethyl-isopropyl-amiloride, HOE 642 and tetrodotoxin to reduce anoxic NE release. Thus, activation of H3R appears to result in an inhibition of both NHE- and voltage-dependent Na+ channels. Most importantly, endogenous histamine was released from the anoxic human heart, and thioperamide and clobenpropit each alone increased NE release, indicating that H3R become activated in myocardial ischemia. Our findings indicate that H3Rs are likely to mitigate sympathetic overactivity in the ischemic human heart and suggest new therapeutic strategies to alleviate dysfunctions associated with myocardial ischemia. PMID- 9353364 TI - Differences in the antinociceptive effects of alpha-2 adrenoceptor agonists in two substrains of Sprague-Dawley rats. AB - In this study, we examined whether Sprague-Dawley rats obtained from two different vendors, Harlan and Sasco, differ with respect to the types of alpha-2 adrenoceptors in the spinal cord that mediate antinociception. This hypothesis was tested using two alpha-2 adrenoceptor agonists, dexmedetomidine and ST-91, which are relatively selective for alpha-2A and alpha-2B adrenoceptors, respectively, and two different measures of nociception, the tail-flick and the 55 degrees C hot-plate test. Dexmedetomidine and ST-91 each increased tail-flick latency to a similar extent in both Harlan and Sasco rats, although dexmedetomidine was more efficacious than ST-91 in each substrain. However, the efficacy of these agonists was markedly different in Harlan and Sasco rats when the hot-plate test was used. For example, ST-91 was a full agonist in the hot plate test in Harlan rats but a weak partial agonist in Sasco rats. Dexmedetomidine was a very weak partial agonist in Harlan rats and ineffective in the hot-plate test in Sasco rats. These findings suggest that (1) both spinal alpha-2A and alpha-2B receptors modulate nociceptive responses in the tail-flick test in both Harlan and Sasco rats; (2) hot-plate responses are mediated predominantly by alpha-2B adrenoceptors, with a minimal contribution by alpha-2A adrenoceptors in the Harlan rat and (3) hot-plate responses are not appreciably affected by either alpha-2A or alpha-2B adrenoceptors in the Sasco rat. These findings confirm previous reports that intrathecal administration of alpha-2 adrenoceptor agonists produces thermal antinociception in the rat. However, the magnitude of the antinociceptive effect is dependent on the receptor selectivity of the agonist used, cutaneous tissue stimulated to elicit nociceptive responses and substrain of rat. PMID- 9353363 TI - Opioid efficacy in a C6 glioma cell line stably expressing the delta opioid receptor. AB - A C6 glioma cell line stably transfected with the rat delta opioid receptor (C6delta) was used to characterize receptor binding and G protein activation by both peptide and nonpeptide delta opioid ligands. The ligand binding affinities for [3H]naltrindole and [3H]pCl-[D-Pen2,D-Pen5]enkephalin (DPDPE) were similar to those observed in monkey brain membranes. The nonpeptide agonists, BW373U86 and SNC80, as well as peptide agonist [D-Ser2, L-Leu5]enkephalyl-Thr maximally stimulated [35S]GTPgammaS binding by 640, 654 and 576%, respectively, over basal. The peptide agonists, DPDPE and deltorphin II, both stimulated [35S]GTPgammaS binding by 375%. Etorphine, diprenorphine, oxymorphindole and 7 spiroindanyloxymorphone were also partial agonists in this assay, although they were less efficacious than deltorphin II. Stimulation of [35S]GTPgammaS binding by agonists was blocked completely by pertussis toxin pretreatment. Both delta-1 and delta-2 selective antagonists 7-benzylidenenaltrexone and a benzofuran analog of naltrindole displayed high affinity for the cloned receptor (0.04 and 0.08 nM) and antagonized the stimulation of [35S]GTPgammaS binding by BW373U86 and DPDPE with similar potencies. Other evidence suggesting the lack of receptor subtypes includes the finding that stimulation of [35S]GTPgammaS binding by receptor subtype selective ligands DPDPE and deltorphin II was not additive. BW373U86, SNC80 and DPDPE maximally inhibited forskolin-stimulated adenylyl cyclase. These cells highly express a homogeneous population of delta opioid receptor that couple to inhibitory Go/Gi proteins. Ligand affinity for the delta opioid receptor correlates with ligand EC50 values for stimulation of [35S]GTPgammaS binding. PMID- 9353365 TI - Mechanism for the nonlinear pharmacokinetics of erythropoietin in rats. AB - The contribution of erythropoietin- (EPO) receptors in target tissues, such as bone marrow and spleen, to the nonlinear pharmacokinetics of recombinant human EPO (rh-EPO) was evaluated in rats. The total body clearance after i.v. administration of rh-EPO (0.2-5 microg/kg) decreased as the dose of rh-EPO increased, approaching a plateau at high doses. The uptake clearance of 125I-rh EPO by the target tissues, bone marrow and spleen, exhibited clear saturation. The Km values ranged from 240 to 450 pM, which are comparable with the reported value for the dissociation constant of EPO binding to EPO-receptors (180 pM) in rat bone marrow cells. A single s.c. administration of a large dose of rh-EPO (1 microg/kg) caused a reduction in tissue uptake clearance of 125I-rh-EPO by bone marrow and spleen (down-regulation). Furthermore, repeated intravenous injection of rh-EPO caused up-regulation of the tissue uptake clearance of 125I-rh-EPO, especially by the spleen, in a dose-dependent manner. Hematopoietic parameters such as hematocrit and hemoglobin concentration were also increased by repeated rh-EPO treatment and significantly correlated with the sum of the tissue uptake clearances in bone marrow and spleen. These findings suggest that the pharmacological receptor could be an important factor in defining the nonlinear pharmacokinetics of rh-EPO. PMID- 9353366 TI - Inhibition of bufalin on pituitary and testicular function in rats. AB - The effects of bufalin on the secretion of testosterone and luteinizing hormone (LH) and the accumulation of testicular adenosine 3':5'-cyclic monophosphate (cAMP) were studied. Male rats were injected with bufalin, human chorionic gonadotropin (hCG), gonadotropin releasing hormone (GnRH), hCG plus bufalin or GnRH plus bufalin via a jugular catheter. Blood samples were collected at several intervals subsequent to the challenge. In the in vitro study, rat testis blocks were incubated with bufalin, hCG or both for 1 h. The anterior pituitary gland was incubated with bufalin, GnRH or both for 30 min. The media were analyzed for testosterone or LH. For studying cAMP accumulation, testicular blocks were incubated for 1 h with the medium containing isobutyl-1-methylxanthine. After incubation, tissues were extracted by ethanol before measuring cAMP concentration. A single intravenous injection of bufalin decreased the basal and hCG-stimulated levels of plasma testosterone. Administration of bufalin in vitro resulted in an inhibition of both basal and hCG-stimulated release of testosterone. Bufalin diminished cAMP accumulation in rat testes. However, the basal levels of plasma and medium LH were not altered by bufalin administration. Likewise, the LH response to GnRH was diminished by bufalin administration, both in vivo and in vitro. These results suggest that the inhibition of testosterone production by bufalin is partly caused by a decrease of testicular cAMP accumulation and LH response to GnRH in rats. PMID- 9353367 TI - Benzyl-polyamines: novel, potent N-methyl-D-aspartate receptor antagonists. AB - The effects of benzyl-polyamines were studied at recombinant N-methyl-D-aspartate (NMDA) receptors expressed in Xenopus laevis oocytes. A number of mono-, di- and tri-benzyl polyamines, having benzyl substitutions on the terminal or central amino groups, inhibited responses of NR1/NR2 receptors in oocytes voltage-clamped at -70 mV. Among the most potent compounds was N1,N4, N8-tri-benzyl-spermidine (TB-3-4), which had an IC50 value of 0.2 microM. TB-3-4 was approximately 40-fold more potent at NR1/NR2A and NR1/NR2B receptors than at NR1/NR2C or NR1/NR2D receptors. Block by TB-3-4 was strongly voltage dependent. Using voltage ramps analyzed by the Woodhull model of voltage-dependent channel block, TB-3-4 was found to have a Kd(0) value of 5 microM and a zdelta value of 1.41 at NR1/NR2B channels, whereas the affinity of binding [Kd(0) = 250 microM] but not the degree of voltage-dependence (zdelta = 1.43) was much lower at NR1/NR2D channels. At a concentration of 10 microM, TB-3-4 had no effect on alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid receptors expressed from the GluR1 subunit, indicating that TB-3-4 is a selective NMDA antagonist. TB-3-4 did not permeate wild-type NMDA channels but could easily permeate channels containing an N616G mutation in the NR1 subunit. This mutation is presumed to increase the size of the narrowest constriction of the NMDA channel, thus allowing passage of TB-3-4. Benzyl-polyamines such as TB-3-4 represent a structurally novel class of NMDA receptor channel blockers. PMID- 9353368 TI - A new milrinone analog: role of binding to A1 adenosine receptor in its positive inotropic effect on isolated guinea pig and rat atria. AB - In electrically driven left atria isolated from guinea pig and rat, a new milrinone analog, 6-ethyl-5-propionyl-1,2-dihydro-2-oxo-3-pyridine carbonitrile, produced a positive inotropic effect that was not dependent on adrenergic mechanisms and was more marked than that exerted by the parent compound. Its inotropic action was almost completely abolished by pretreatment of atria with adenosine deaminase and correlated well with its binding ability to the cardiac adenosine A1 receptor. In this regard, the analog showed a 100-fold higher affinity for adenosine receptor than that of milrinone. Moreover, it shifted to the right the concentration-response curves for the negative inotropic action of the stable adenosine receptor agonist R-phenylisopropyladenosine. The new analog behaved as a competitive inhibitor of Type III phosphodiesterase isolated from both guinea pig and rat, although its Ki value was 10 times higher than that of milrinone. However, an increase in cAMP levels does not seem to be involved in the mechanism of action of the new compound, because the presence of carbachol did not decrease the extent of the positive inotropic effect of the analog and did not modify its EC50 in either guinea pig or rat myocardial preparations. Taken together, these results suggest that the milrinone structure can be modified, giving rise to a more active compound whose inotropic effect in both guinea pig and rat appears to be more clearly related to antagonism toward endogenous adenosine than to Type III phosphodiesterase inhibition. PMID- 9353369 TI - Diadenosine polyphosphates directly relax porcine coronary arterial smooth muscle. AB - By use of front-surface fluorometry and fura-2-loaded medial strips of the porcine coronary artery, cytosolic Ca++ concentration ([Ca++]i) and force development were monitored simultaneously to determine the mechanisms of vasorelaxation induced by the diadenosine polyphosphates (APnA) diadenosine 5',5'''-P1, P4-tetraphosphate (AP4A) and diadenosine 5',5'''-P1,P5-pentaphosphate (AP5A). APnA concentration-dependently inhibited the sustained elevations of [Ca++]i and force induced by U-46619, a thromboxane A2 analog, in the presence of extracellular Ca++. APnA shifted the [Ca++]i-force relation curves of contractions induced by various concentrations of high K+ to the right. The AP4A induced decreases in [Ca++]i and force were largely attenuated by tetrabutylammonium. The AP4A-induced decreases in force were attenuated by 4 aminopyridine and charybdotoxin. The AP5A-induced decreases in [Ca++]i and force were attenuated by tetrabutylammonium, 4-aminopyridine and charybdotoxin. In the absence of extracellular Ca++, APnA did not inhibit the transient elevations of [Ca++]i induced by histamine or caffeine. Both AP4A and AP5A increased intracellular cAMP content. We thus conclude that AP4A and AP5A relax the porcine coronary artery by decreasing [Ca++]i, possibly through the activation of K+ channels, but not through inhibition of intracellular Ca++ release and by decreasing the Ca++ sensitivity of the contractile machinery. These effects were considered to be mediated by cAMP. PMID- 9353370 TI - The role of dopaminergic systems in opioid receptor desensitization in nucleus accumbens and caudate putamen of rat after chronic morphine treatment. AB - Morphine treatment of rats (60-70 mg/kg/day, 7 days) reduced delta opioid receptor-mediated inhibition of adenylyl cyclase activity in caudate putamen without any change in regulation by mu receptors. Earlier studies suggested that dopamine D1 and mu opioid receptors that regulate adenylyl cyclase are expressed preferentially by striato-nigral neurons, whereas adenosine A2a and delta1 opioid receptors are expressed preferentially by striato-pallidal neurons. Chronic morphine treatment also resulted in a reduction of dopamine D2 receptor-mediated inhibition of A2a receptor-stimulated adenylyl cyclase. Treatment with a D2 receptor antagonist (eticlopride; 1 mg/kg/day) for 7 days reduced D1 receptor stimulation of adenylyl cyclase. In contrast, chronic treatment with a D1 receptor antagonist R(+)-7-chloro-8-dihydroxy-3-methyl-1-phenyl-2,3,4, 5 tetrahydro-1H-3-benzazepine HCL (SCH 23390; 2.5 mg/kg/day) resulted in a reduction of delta1 and delta2 opioid inhibition of adenylyl cyclase, with no change in the inhibitory activity of a mu agonist. The inhibitory activity of the D2 agonist quinelorane against adenosine A2a-activated enzyme was also reduced by this treatment. Thus chronic D1 blockade, like chronic morphine treatment, appears to cause a selective impairment of the regulation of adenylyl cyclase in A2a receptor-expressing striato-pallidal neurons. D2 receptor activation appears to play an important role in the desensitization of delta receptors, because concurrent administration of the D2 antagonist eticlopride with morphine prevented the densitization of delta and D2 receptors. Similar results were obtained in nucleus accumbens, which suggests a role for D2 receptor desensitization in the adaptive response of this brain region to chronic morphine. PMID- 9353371 TI - Characterization of the discriminative stimulus produced by the dopamine antagonist tiapride. AB - The ability of tiapride, a selective D2/D3 dopamine receptor antagonist, to exert discriminative stimulus control of responding was investigated by training rats to discriminate this drug (30 mg/kg) from saline in a two-lever, food reinforcement procedure. Acquisition of tiapride discrimination required a relatively lengthy training period (mean of 76 sessions) but stable performance was maintained throughout the 18- month study. The dose of tiapride eliciting 50% tiapride-lever choice (ED50) was 2.2 mg/kg. After determination of the dose effect curve with tiapride, substitution tests with several dopamine antagonists and other reference compounds were performed. All dopamine antagonists, including amisulpride (ED50 4 mg/kg), sulpiride (18 mg/kg), sultopride (1.5 mg/kg), clebopride (0.13 mg/kg), raclopride (0.16 mg/kg), metoclopramide (1.4 mg/kg), remoxipride (4.8 mg/kg), pimozide (2.7 mg/kg), thioridazine (3.4 mg/kg), olanzapine (0.97 mg/kg), chlorpromazine (1.9 mg/kg), risperidone (0.22 mg/kg) and haloperidol (0.14 mg/kg), except clozapine (>10 mg/kg), produced dose-dependent substitution for tiapride. Tiapride-like stimulus effects were observed at doses that decreased response rates. However, ED50 values for substitution by tiapride, amisulpride, sulpiride, sultopride, pimozide, clebopride and thioridazine were lower than ED50 values for decreasing responding. Additional studies were conducted to evaluate the ability of direct and indirect dopamine agonists to attenuate the tiapride discriminative stimulus. Pretreatment with d-amphetamine and nomifensine antagonized the discriminative stimulus effects of tiapride. Quinpirole, 7-OH-DPAT, bromocriptine and apomorphine partially blocked the stimulus effects of tiapride whereas SKF 38393 did not affect the discrimination. These results from substitution and antagonism tests indicated that the discriminative effects of tiapride are mediated by activity at D2/D3 dopamine receptors. PMID- 9353372 TI - P-Glycoprotein mediates the efflux of quinidine across the blood-brain barrier. AB - Recent studies suggest that P-glycoprotein located on the blood-brain barrier restricts the brain uptake of its substrates. We examined the role of P glycoprotein on the restricted entry of quinidine to the brain. Quinidine is a well known inhibitor of P-glycoprotein, although it is not yet clarified whether quinidine is the substrate for P-glycoprotein. Kinetic analysis of the uptake of quinidine into the rat brain after intravenous bolus administration revealed that the net uptake clearance is 25.5 microl/min/g brain. Intravenous administration of SDZ PSC 833, a multidrug resistance modifier, enhanced the net uptake clearance of quinidine by 15.7-fold. In contrast, no enhancement by SDZ PSC 833 was observed for the brain uptake of mannitol, a marker for the passive diffusion across the blood-brain barrier. The elimination of [3H] quinidine from the rat brain after microinjection into the cerebral cortex was inhibited by preadministered unlabeled quinidine and verapamil. In addition, the brain-to plasma concentration ratio of quinidine at 10 min after intravenous administration was 27. 6-fold higher in mdr1a knock-out mice than in control mice. These results suggest that P-glycoprotein mediates the efflux of quinidine across the blood-brain barrier, resulting in its restricted entry to the brain. PMID- 9353373 TI - In vivo criteria to differentiate monoamine reuptake inhibitors from releasing agents: sibutramine is a reuptake inhibitor. AB - Because monoamine reuptake inhibitors and releasing agents both increase extracellular neurotransmitter levels, establishing in vivo experimental criteria for their classification has been difficult. Using microdialysis in the hypothalamus of unanesthetized rats, we provide evidence that serotonin- (5-HT) selective and nonselective reuptake inhibitors can be distinguished from the 5-HT releasing agent fenfluramine by four criteria: 1) Systemic fenfluramine produces a much greater increase in 5-HT than the reuptake inhibitors. 2) The 5-HT somatodendritic autoreceptor agonist, (+/-)-8-hydroxy-(dipropylamino)tetralin (8 OH-DPAT), attenuates the increase in 5-HT produced by reuptake inhibitors, but not by fenfluramine. 3) The large increase in 5-HT produced by infusion of reuptake inhibitors into the hypothalamus is attenuated by their systemic administration. However, systemic injection of fenfluramine during its local infusion does not attenuate this increase. 4) Reuptake inhibitor pretreatment attenuates fenfluramine-induced increases in 5-HT. According to these criteria, the in vivo effects of the novel antiobesity drug sibutramine are consistent with its characterization as a 5-HT reuptake inhibitor and not a 5-HT releaser. Thus, sibutramine produced increases in hypothalamic 5-HT similar in magnitude to the effects of the known reuptake inhibitors, and the increase was attenuated by 8-OH DPAT. Also, sibutramine attenuated fenfluramine-induced 5-HT release. Systemic administration of sibutramine failed to attenuate the increase in 5-HT produced by its local infusion, suggesting that this criterion is not applicable to compounds with low affinity for the 5-HT transporter. PMID- 9353374 TI - Effects of proposed treatments for cocaine addiction on hemodynamic responsiveness to cocaine in conscious rats. AB - Several agents may treat cocaine addiction and toxicity including bromocriptine, desipramine, GBR 12909 [1-(2-(bis(4-fluorphenyl)-methoxy)-ethyl)-4-(3-phenyl propyl) piperazine], diazepam, buprenorphine and dizocilpine. In this study, we sought to determine whether these specific therapeutic agents alter cardiovascular responses to cocaine in conscious rats. Arterial pressure responses to cocaine (5 mg/kg, i.v.) were similar in all rats whereas cardiac output responses varied widely. In 26 of 33 rats (named vascular responders), cocaine induced a decrease in cardiac output of 8% or more. The remaining rats with little change or an increase in cardiac output were classified as mixed responders. Pretreatment with bromocriptine (0.1 mg/kg) or desipramine (1 mg/kg) increased cardiac output in mixed responders and increased systemic vascular resistance in vascular responders similar to the differential effects noted with cocaine. GBR 12909 (0.5-10 mg/kg) elicited a decrease in cardiac output at higher doses. Diazepam (0.1 and 0.5 mg/kg) had small, short-lasting effects on cardiovascular parameters. Buprenorphine (0.3 mg/kg) or the NMDA (N-methyl-D aspartic acid) receptor antagonist, dizocilpine (0.05 mg/kg), increased arterial pressure, heart rate and cardiac output in vascular responders. Bromocriptine and desipramine prevented the difference in cardiac output responses in vascular and mixed responders by reducing the cocaine-induced decrease in cardiac output in vascular responders. Pretreatment with GBR 12909 (1 mg/kg) had little effect on cardiovascular responses to cocaine except to depress the increase in cardiac output noted in mixed responders. Buprenorphine selectively enhanced the increase in systemic vascular resistance whereas dizocilpine enhanced the pressor response. These data suggest that several treatment regimens for cocaine addiction alter the cardiovascular responses to cocaine and that dopamine D2 receptor activation may be necessary for the decrease in cardiac output noted in vascular responders. PMID- 9353375 TI - Nonopioid motor effects of dynorphin A and related peptides: structure dependence and role of the N-methyl-D-aspartate receptor. AB - Dynorphin (Dyn) A and related opioid and nonopioid peptides were tested for their ability to produce motor effects in mice. Central (intracerebroventricular) administration of Dyn A in mice produced marked motor effects characterized by wild running, jumping, circling and/or barrel rolling with an ED50 value of 14.32 (95% confidence limits, 10.09-20.32) nmol/mouse. The order of potency of the various Dyn A-related peptides and fragments in producing motor effects was Dyn A approximately Dyn A-(1-13) > [Ala1]Dyn A-(1-13) approximately Dyn A-(2-13) > alpha-Neo-End > Dyn A-(1-8) approximately Dyn B approximately Dyn A-(2-8) >>> Dyn A-(3-8). Dyn A-(1- 5) (or Leu-Enk) and Dyn A-(6-10) displayed no motor effect at doses up to 100 nmol/mouse. The potencies of Dyn A and Dyn A-(2-13) were not affected by preadministration of naloxone (5 mg/kg s.c.), but the motor effects of Dyn A-(1-13) (20 nmol/mouse i.c.v.) were significantly reduced by coadministration of low doses (0.2-0.6 nmol/mouse) of the N-methyl-D-aspartate (NMDA) receptor antagonists dextrorphan, MK-801 and CPP. Dyn A was also a potent inhibitor of the binding of the phencyclidine receptor ligand, [3H]MK-801, to rat brain membranes, with a Ki value of 0.41 microM. However, the order of potency of the various Dyn A-related peptides and fragments in inhibiting [3H]MK-801 binding did not correlate with their ability to produce motor effects. On the other hand, Dyn A and related peptides produced a significant potentiation of the binding of the competitive NMDA antagonist [3H]CGP-39653 to rat brain membranes, an effect that correlated well (r = 0.91) with their potency in producing motor effects. These results indicate that the nonopioid motor effects of Dyn A and related peptides are structure dependent, with Dyn A-(2-8) being the minimal core peptide for motor activity. In addition, these effects most likely involve the participation of the excitatory amino acid binding domain on the NMDA receptor complex. PMID- 9353376 TI - Gastrointestinal absorption of recombinant human insulin-like growth factor-I in rats. AB - The GI absorption of recombinant human insulin-like growth factor-I (rhIGF-I) and its improvement were investigated in rats. The 125I-rhIGF-I rapidly degraded to the trichloroacetic acid-soluble form in the small-intestinal contents, but it was relatively stable in the gastric and large-intestinal contents and in the subcellular fraction of the small-intestinal mucosa. To protect rhIGF-I from degradation in the small-intestinal contents, the effect of some adjuvants was examined and their degradation was markedly inhibited by the presence of aprotinin or casein. After p.o. administration of 125I-rhIGF-I at the dose of 1.0 mg/kg, trichloroacetic acid-precipitable radioactivity in the plasma was periodically determined. We found that a considerable amount of rhIGF-I was absorbed into the systemic circulation and that the bioavailability was 9.3%, which is much greater than that of insulin. The coadministration of aprotinin and that of casein enhanced the bioavailability further: 46.9% and 67.0%, respectively. Radioimmunoassay using a monoclonal antibody for rhIGF-I confirmed the high bioavailability of immunoreactive rhIGF-I. From gel chromatography of plasma, the radioactivity in the plasma was found to be in the form of high molecular-weight complexes. The mechanism for the uptake of rhIGF-I by intestinal mucosa may be absorptive-mediated endocytosis. PMID- 9353377 TI - Phosphodiesterase isoforms in the pulmonary arterial circulation of the rat: changes in pulmonary hypertension. AB - Phosphodiesterase (PDE) activity was determined in pulmonary arteries removed from control and chronic hypoxia-induced pulmonary hypertensive rats. The main, first-branch, intrapulmonary and resistance pulmonary arteries were studied. We measured total cAMP PDE activity and cGMP PDE activity, as well as that of individual isoforms (PDE1-5). cAMP PDE activity in chronic hypoxic rats was increased in first-branch and intrapulmonary arteries from hypoxic rats. No changes were observed in the main or resistance pulmonary arteries. Similarly, cGMP PDE activity was increased in the main, first-branch and intra-pulmonary arteries of the hypoxic rats. No changes in cGMP PDE activity were observed in resistance arteries. There was evidence for PDE1-5 activity in all pulmonary arteries. The increased cAMP PDE activity in first-branch and intrapulmonary vessels was associated with an increase in cilostimide-inhibited PDE (PDE3) activity. Increased total cGMP PDE in main pulmonary artery was associated with increases in Ca++/calmodulin-stimulated (PDE1) activity. An increase in zaprinast inhibited (PDE5) activity was observed in first-branch and intrapulmonary arteries. Our results suggest that decreases in intracellular cyclic nucleotide levels in pulmonary arteries from pulmonary hypertensive rats are associated with increased PDE activity. Further, these changes may reflect alterations at the level of specific types of PDE isoforms. PMID- 9353378 TI - Beneficial effects of long-term enalapril treatment and low-salt intake on survival rate of dahl salt-sensitive rats with established hypertension. AB - We investigated the effects of long-term treatment with the angiotensin converting enzyme inhibitor enalapril and low-salt intake on the survival rate of Dahl salt-sensitive rats fed a high-salt (6.0% NaCl) diet. The systolic blood pressure of the rats increased gradually from 5 weeks of age and reached >240 mm Hg at 12 weeks of age. At this point, a low-salt diet group received a placebo (group 1, n = 10), and the high-salt diet group was divided into three groups: those given a placebo with the high-salt diet (group 2, n = 15), those given a chow change from a high- to a low-salt diet with a placebo (group 3, n = 14) and those given enalapril (30 mg/kg/day p.o., group 4, n = 14). At 19 weeks of age, all rats in group 1 were alive, and the survival rate of group 2 was only 40% (P < .001 vs. group 1). The survival rates of both groups 3 and 4 were significantly better: 86% (P < .01 vs. group 2) and 93% (P < .01), respectively. This beneficial effect on mortality was accompanied by an amelioration of the elevated plasma creatinine and urea nitrogen levels and a decrease in the glomerular sclerosis lesion scores in both groups. These results suggested that a high-salt content diet and the renin-angiotensin system are deterioration factors in lethal renal damage and the limitation of the diet salt content and inhibition of the renin-angiotensin system are important to improve the survival rate in high-salt loaded hypertensive Dahl salt-sensitive rats. PMID- 9353379 TI - Exacerbation of methamphetamine-induced neurochemical deficits by melatonin. AB - Methamphetamine (METH), administered in large, repeated doses, compromises the dopaminergic and serotonergic systems as indicated by prolonged suppression of tyrosine hydroxylase and tryptophan hydroxylase activity and concurrent decreases in the content of dopamine and 5-hydroxytryptamine. Because dopamine is necessary for these dopaminergic and serotonergic deficits we postulated that dopamine and/or its reactive metabolites are responsible for these degenerative alterations. Because we previously demonstrated that in vitro reducing conditions reverse the decrease in tryptophan hydroxylase activity, we reasoned that melatonin, a purported endogenous antioxidant, may alter this response. Rats were treated with METH and/or melatonin and trytophan hydroxylase activity and 5 hydroxytryptamine content were assessed; tyrosine hydroxylase activity and dopamine content were also measured. Not only did melatonin not prevent METH induced deficits in serotonergic and dopaminergic parameters, but coadministration of melatonin with METH actually enhanced most of the monoaminergic effects of METH. This enhancing effect could not be attributed to alteration of body temperature. Because METH abuse causes insomnia and melatonin is promoted in some countries for insomnia, the implications of the interaction between these two drugs could be clinically important. PMID- 9353381 TI - d-Methadone is antinociceptive in the rat formalin test. AB - The l-isomer of methadone possesses opioid activity, whereas the d-isomer is weak or inactive as an opioid. Both d- and l-methadone have been shown to bind to the N-methyl-D-aspartate (NMDA) receptor. To determine whether d-methadone has functional, in vivo NMDA receptor antagonist activity, the antinociceptive effects of d-methadone were evaluated in the rat tail-flick and formalin tests. Cumulative dose-response analysis in the tail-flick test revealed an ED50 value for intrathecal (spinal) l-methadone of 15.6 microg/rat. In contrast, spinal d methadone produced no antinociception at a cumulative dose of 460 microg/rat. d Methadone in a dose range from 32 to 320 microg/rat dose-dependently reduced formalin-induced flinching behavior during phase 2 but not during phase 1 of the formalin test. These antinociceptive effects of d-methadone were not blocked by a spinal dose of naloxone that effectively antagonized an antinociceptive (tail flick test) dose of l-methadone. d-Methadone at an intrathecal dose of 250 microg shifted the ED50 value for NMDA-induced nociceptive behaviors more than 3-fold to the right, which indicates an antagonism of these NMDA receptor-mediated effects. These results indicate that d-methadone is antinociceptive as a result of its NMDA receptor antagonist activity. PMID- 9353380 TI - Biological profile of L-745,870, a selective antagonist with high affinity for the dopamine D4 receptor. AB - L-745,870,(3-([4-(4-chlorophenyl)piperazin-1-yl]methyl)-1H- pyrollo[2,3-b] pyridine, was identified as a selective dopamine D4 receptor antagonist with excellent oral bioavailability and brain penetration. L-745,870 displaced specific binding of 0.2 nM [3H] spiperone to cloned human dopamine D4 receptors with a binding affinity (Ki) of 0. 43 nM which was 5- and 20-fold higher than that of the standard antipsychotics haloperidol and clozapine, respectively. L 745,870 exhibited high selectivity for the dopamine D4 receptor (>2000 fold) compared to other dopamine receptor subtypes and had moderate affinity for 5HT2, sigma and alpha adrenergic receptors(IC50 < 300 nM). In vitro, L-745,870 (0.1-1 microM) exhibited D4 receptor antagonist activity, reversing dopamine (1 microM) mediated 1) inhibition of adenylate cyclase in hD4HEK and hD4CHO cells; 2) stimulation of [35S] GTPgammaS binding and 3) stimulation of extracellular acidification rate, but did not exhibit any significant intrinsic activity in these assays. Although standard antipsychotics increase dopamine metabolism or plasma prolactin levels in rodents, L-745,870 ( 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) > hexahydrosiladiphenidol hydrochloride (HHSiD) > tripitramine > pirenzepine > AF DX-116 > methoctramine) as well as their postjunctional affinity estimates (pA2) are in keeping with the notion that muscarinic receptors responsible for bladder contraction belong to the M3 subtype. The M3 subtype-preferring 4-DAMP and HHSiD did not discriminate between prejunctional and postjunctional effects. The M2/M4 subtype-preferring antagonists tripitramine, methoctramine and AF-DX 116 were more potent in facilitating the evoked [3H]ACh release than in inhibiting the contractile response. The rank order of prejunctional potencies was atropine > 4 DAMP > tripitramine > HHSiD > methoctramine > AF-DX 116 > pirenzepine, indicating the involvement of M4 receptors. Furthermore, when potency relationship was determined by correlating prejunctional-log EC50 values with published constants for cloned and natives muscarinic receptor subtypes, the correlations were significant for both M4 and M5 subtypes, but the best correlation found (P < .001) was for the M4 subtype. These findings suggest that the negative feedback mechanism inhibiting the release of ACh in the rat urinary bladder is mediated by prejunctional autoreceptors of the M4 subtype. PMID- 9353396 TI - Modafinil induces wakefulness without intensifying motor activity or subsequent rebound hypersomnolence in the rat. AB - Modafinil, a novel compound for treating excessive sleepiness, potently increases wakefulness in laboratory rodents, cats, monkeys and humans. Although its mechanism of action is unknown, modafinil appears to be unlike classic stimulants. We investigated this generality by testing the selectivity of this compound for wake-promoting effects (e.g., relative to locomotor effects) and homeostatic sleep responses after drug-induced waking relative to the prototypical stimulant methamphetamine (METH). Continuous measures of electroencephalogram (EEG) sleep-wakefulness, locomotor activity (LMA) and body temperature (Tb) were obtained from adult male Wistar rats 3 days before and after treatment with modafinil (30, 100 and 300 mg/kg i.p.), 0.25% methylcellulose (vehicle) or METH (0.5 and 1.0 mg/kg i.p.). Individually housed rats in a 24-h light-dark cycle (LD 12:12) were treated 5 h after lights-on (CT 5). LMA and Tb were monitored via intraperitoneal telemetry. Sleep-wake stages and LMA were recorded every 10 s, Tb every minute. During the first 3 h post treatment, modafinil and METH significantly and dose-dependently increased EEG wake time (P < .01 for 30 mg/kg modafinil, all other P < .0001) and wake episode duration. Although the cumulative increases in wakefulness were statistically equivalent, METH, but not modafinil, produced subsequent rebound hypersomnolence. At these equipotent wake-promoting doses, modafinil produced the same total amount of REM sleep inhibition but during a longer time than METH. Modafinil also increased LMA amount (counts/h, P < .001) and LMA intensity (counts/min awake, P < .001) less than METH. Both rebound hypersomnolence and increased LMA intensity, which are undesirable features in wake-promoting drugs, were not observed after modafinil treatment, and thus further differentiated modafinil from amphetamine like stimulants. PMID- 9353397 TI - Activation of cardiac ATP-sensitive K+ channels by KRN4884, a novel K+ channel opener. AB - In the present study, we have investigated the mechanism underlying the activation by 5-amino-N-[2-(2-chlorophenyl)ethyl]-N'-cyano-3- pyridinecarboxamidine (KRN4884), a new K+ channel opener, of ATP-sensitive K+ (KATP) channels in single ventricular cells of guinea pig hearts by the inside out patch-clamp method. In the presence of intracellular ATP (1 mM), KRN4884 (0.1 3 microM) activated KATP channels in a concentration-dependent manner (EC50 = 0.55 microM) without affecting the unitary current conductance and the gating properties. KRN4884 (0.3 microM) shifted the concentration-response relationship for ATP-induced KATP channel inhibition to the right and slightly upward direction without altering the slope. After either the spontaneous or Ca++ induced channel rundown, KRN4884 (1 and 3 microM) partially restored the KATP channel activity. Furthermore, the effect of KRN4884 was augmented by the presence of uridine 5'-diphosphate (3 mM). The results indicate that KRN4884 activates cardiac KATP channels through not only decreasing the sensitivity of the channel to ATP but also directly stimulating the opening of the channel. PMID- 9353398 TI - Nicotinic-receptor mediation of S(-)nornicotine-evoked -3H-overflow from rat striatal slices preloaded with -3H-dopamine. AB - Previous results from our laboratory demonstrated that S(-)nornicotine, a major tobacco alkaloid and an active nicotine metabolite present in the CNS, increases dopamine release from rat striatal slices in a concentration-dependent and calcium-dependent manner. The present study determined if S(-)nornicotine-evoked dopamine release was the result of nicotinic receptor stimulation. Stereoselectivity and the ability of classical noncompetitive and competitive nicotinic receptor antagonists (mecamylamine (MEC) and dihydro-beta-erythroidine (DHbetaE), respectively) to inhibit S(-)nornicotine-evoked [3H]overflow from [3H]dopamine-preloaded rat striatal slices were investigated. Nornicotine increased [3H]overflow in a stereoselective manner at concentrations from 1 to 100 microM. MEC (0.01-100 microM) or DHbetaE (0.01-10 microM) alone did not evoke -3H-overflow. However, 100 microM DHbetaE evoked -3H-overflow, and therefore, was not used in experiments investigating antagonism of S(-)nornicotine's effect. MEC and DHbetaE inhibited S(-)nicotine- (10 microM) evoked [3H]overflow in a concentration-dependent manner. Concentrations of MEC (100 microM) and DHbetaE (10 microM) which maximally inhibited S(-)nicotine's effect were chosen for subsequent experiments determining inhibition of the effect of S(-)nornicotine (0.1 microM-3 mM). MEC and DHbetaE significantly inhibited the effect of low concentrations (<100 microM) of S(-)nornicotine; however, higher concentrations (>100 microM) of S(-)nornicotine were not inhibited by either nicotinic antagonist. Taken together, the results suggest that low concentrations of S( )nornicotine stimulate nicotinic receptors to evoke the release of dopamine from dopaminergic presynaptic terminals. Thus, nornicotine, which acts as an agonist at neuronal nicotinic receptors, may contribute to the neuropharmacological effects of nicotine and tobacco use. PMID- 9353399 TI - Bronchoconstrictor and respiratory effects of neurokinin A in dogs. AB - Neurokinin A (NKA) is the primary bronchoconstrictor tachykinin in the lungs of several species, including humans and has been implicated as an important mediator of inflammatory lung disorders, such as asthma. In this study, we investigated the effect of NKA on airway mechanics (lung resistance, dynamic lung compliance) and respiration (tidal volume, respiratory rate) in anesthetized, spontaneously breathing, male beagle dogs. The dogs were challenged with aerosolized NKA that was delivered from a jet nebulizer to the airways through an endotracheal tube. The challenge consisted of five separate inflations of 600 ml of air/inflation over a 1-min period. Challenge with aerosolized NKA (0.1-1%) produced a dose-dependent increase in lung resistance and a decrease in dynamic lung compliance. The bronchoconstriction induced by 1% NKA peaked at 0.5 min after challenge and had a duration of approximately 5 min. Challenge with 1% NKA also reduced tidal volume and increased respiratory rate. Pretreatment of dogs with the NK-2 receptor antagonist, SR 48968 dose-dependently (1-10 mg/kg, p.o.) blocked the bronchoconstriction and respiratory responses to NKA challenge. Pretreatment with the NK1-receptor antagonist, CP 99994 (1 mg/kg, i. v.) had no effect on the increase in lung resistance and the decrease in dynamic lung compliance due to NKA challenge, but blunted the respiratory response to NKA. Pretreatment of dogs with inhaled ipratropium bromide (0.01%) slightly, but significantly reduced the increase in lung resistance due to NKA challenge but had no effect on the decrease of dynamic lung compliance or on the respiratory responses to NKA. As expected, the bronchoconstrictor response to inhaled methacholine was completely blocked by inhaled ipratropium bromide (0.01%). In conclusion, we have identified an NK2-receptor mediated bronchoconstrictor effect of NKA in dogs. Cholinergic reflexes play a small, but significant role in this response. Furthermore, both NK1 and NK2-receptors appear to be involved with the development of the rapid, shallow breathing response to NKA challenge. These results demonstrate an effect of tachykinins on airway mechanics and ventilatory reflexes in dogs. PMID- 9353401 TI - 8-Alkylamino-substituted analogs of N6-cyclopentyladenosine are partial agonists for the cardiovascular adenosine A1 receptors in vivo. AB - Partial adenosine A1 receptor agonists with reduced intrinsic activity at the cardiovascular system would be promising for therapeutic application (e.g., as antilipolytic agents). In the present study a series of 8-alkylamino [methyl (M) , ethyl (E)-, propyl (P)-, butyl (B)- and cyclopentyl (CP)-] derivatives of N6 cyclopentyladenosine (CPA) were investigated in conscious normotensive rats. After intravenous administration of the compounds to rats, heart rate (HR) and mean arterial pressure were monitored continuously, and serial arterial blood samples were drawn for determination of the pharmacokinetics. The concentration heart rate relationships of the compounds were described on the basis of an integrated pharmacokinetic-pharmacodynamic model. The blood concentration-time profiles of the compounds could be described best by a biexponential function. The derivatives of CPA had uniform pharmacokinetic properties. The larger volume of distribution at steady state of the 8-substituted analogs resulted in terminal half-lives (ranging from 17 to 24 min) which were significantly longer than for CPA (7 min). All derivatives of CPA produced less pronounced reductions in HR and MAP than CPA. The relationship between concentration and the reduction in HR was adequately described by the sigmoidal Emax model in individual rats given 8MCPA, 8ECPA and 8PCPA. 8BCPA and 8CPCPA were nearly inactive on heart rate. The in vivo EC50,u values for the reduction in HR (366 nM, 210 nM, 170 nM and 175 nM for 8MCPA, 8ECPA, 8PCPA and 8BCPA, respectively) were in the same order of magnitude as the affinities in receptor binding studies. The order of magnitude of the intrinsic activities (Emax) was CPA > 8MCPA > 8ECPA = 8PCPA > 8BCPA > 8CPCPA, which indicated partial agonism of the compounds in vivo. The in vivo parameter Emax correlated highly (r = 0.97) to the GTP shift observed in radioligand binding experiments. PMID- 9353400 TI - Motor response to a dopamine D3 receptor preferring agonist compared to apomorphine in levodopa-primed 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine monkeys. AB - The profile of dopamine receptor subtype activation contributing to the therapeutic efficacy and motor response complications of levodopa (nonselective pro-agonist) in Parkinson's disease remains unclear. Potent, selective, short acting dopamine D2 receptor subfamily agonists show good antiparkinsonian efficacy but produce dyskinesias comparable to levodopa. Nonetheless, agonists displaying higher affinity for dopamine receptors other than the D2 subtype may have a better therapeutic index. To clarify this issue, we compared the nonselective dopamine D1/D2 receptor subfamilies agonist apomorphine to the dopamine D3 receptor preferring agonist [R-(+)-trans-3,4,4a,10b-tetrahydro-4 propyl-2H,5H-[1]benzopyrano[4 , 3-b]-1,4-oxazin-9-ol] (PD 128,907) in 6 levodopa primed , 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned parkinsonian monkeys with reproducible dyskinesias. Single s.c. dosing with the lowest fully effective dose of apomorphine (averaging 27.9 +/- 4.5 microg/kg) and PD 128,907 (averaging 41.7 +/- 4.4 microg/kg) yielded equivalent antiparkinsonian efficacy on the behavioral scale and portable activity monitoring used. A comparable significant dose-dependent increase in the response magnitude and duration was seen with two higher doses. The severity of dyskinesia was also similar between the two drugs. When the lower dose for each drug was administered six times at a fixed 90-min interval, both drugs remained efficacious with no significant tolerance observed. The D3 receptor preferring antagonist U-99194A significantly reduced the motor effects of both apomorphine and PD 128,907. Thus, increased D3 receptor tone does not acutely ameliorate dyskinesias in levodopa-primed parkinsonian monkeys. Given the reported lack of affinity of PD 128,907 for central D1 receptors, our data support the concept that the pharmacological activation of D1 receptors is not mandatory for relief of parkinsonism and production of dyskinesia. PMID- 9353402 TI - Mechanism-based pharmacokinetic-pharmacodynamic modeling of the effects of N6 cyclopentyladenosine analogs on heart rate in rat: estimation of in vivo operational affinity and efficacy at adenosine A1 receptors. AB - We have developed a pharmacokinetic-pharmacodynamic strategy based on the operational model of agonism to obtain estimates of apparent affinity and efficacy of N6-cyclopentyladenosine (CPA) analogs for the adenosine A1 receptor mediated in vivo effect on heart rate in the rat. All analogs investigated produced a significant decrease of the heart rate after intravenous infusion. Individual concentration-effect curves were fitted to the operational model of agonism with the values of Emax and n constrained to the intrinsic activity (273 bpm) and Hill slope (1.18), respectively, obtained with the agonist that displayed the highest intrinsic activity, 5'-deoxy-CPA. In all cases, the model converged and estimates of apparent affinity and efficacy were obtained for each agonist. Affinity estimates correlated well with pKi values for the adenosine A1 receptor in rat brain homogenates. In addition, a highly significant correlation was found between the estimates of the in vivo efficacy parameter and the GTP shift (the ratio between Ki in the presence and absence of GTP). In conclusion, the operational model of agonism can provide meaningful measures of agonist affinity and efficacy at adenosine A1 receptors in vivo. The model should be of use in the development of partial adenosine A1 receptor agonists. PMID- 9353403 TI - Comparison of the metabolism and toxicity of dapsone in rat, mouse and man. AB - The metabolism and toxicity of dapsone was compared in vitro and in vivo in rat, mouse and man. Metabolism was assessed by high-pressure liquid chromatography mass spectrometry and methemoglobin formation has been used as a toxic endpoint. The greatest toxicity in vitro was seen in microsomes prepared from male Wistar rats (36.6 +/- 1.5% methemoglobin), although toxicity was also seen in microsomes from the female rat (8.2 +/- 1.3%), male CD1 (4.2 +/- 1.6%) and human (10. 9 +/- 1.1%). The rank order of toxicity agreed with the formation of the hydroxylamine metabolite in vitro. All microsomes were also capable of catalyzing the reverse reaction, i.e., reduction of the hydroxylamine to dapsone. However, in vivo administration of dapsone resulted in significant (P < 0.05) methemoglobinemia only in male rats and humans. This species difference in the susceptibility to dapsone toxicity could not be attributed solely to the sensitivity of the target erythrocytes, because the order of sensitivity to dapsone hydroxylamine was human > mouse > rat. Analysis of bile and urine revealed the formation of dapsone hydroxylamine and its glucuronide in male rats and humans, but not in female rats or mice. This species difference in the metabolism and toxicity of dapsone has important implications in the safety evaluation of related compounds for man. PMID- 9353404 TI - Biphasic effect-time courses in man after formoterol inhalation: eosinopenic and hypokalemic effects and inhibition of allergic skin reactions. AB - The kinetics of inhaled racemic formoterol and its effects on the size of the early cutaneous reaction to intradermal injection of an allergen, eosinopenia and hypokalemia were assessed by pharmacokinetic-pharmacodynamic modeling. After inhalation of either 120 microg of formoterol or placebo, blood samples were taken and skin tests were performed in seven healthy subjects. A two-compartment model was needed to describe the observed formoterol plasma concentration-time curves. To describe the observed biphasic concentration, two absorption routes with different absorption rate constants were incorporated in the model. These two phases were explained by rapid absorption via the respiratory tract together with a slower and delayed oral absorption. For the description of the concentration-effect relations, an Emax (the maximum obtainable effect) formula for competitive agonism, with an effect compartment, had to be used. Fitting the wheal and flare, an apparent diurnal variation had to be taken into account by incorporating in the model rising base-line values. For the flare responses, influence of the location on the forearm appeared to be operative. Systemic formoterol absorbed via the oral route behaved differently from the fraction absorbed via the lungs, with EC50 (steady state concentration that gives 50% of maximum effect) values for all three systemic effects being three times lower after oral absorption than after absorption via the respiratory tract. Pharmacodynamic parameters can probably only be estimated quantitatively when the kinetics of the separate enantiomers of formoterol can be taken into account. PMID- 9353405 TI - l-alpha-Acetylmethadol, l-alpha-acetyl-N-normethadol and l-alpha-acetyl-N,N dinormethadol: comparisons with morphine and methadone in suppression of the opioid withdrawal syndrome in the dog. AB - l-alpha-Acetyl-N-normethadol (nor-LAAM) and l-alpha-acetyl-N, N-dinormethadol (dinor-LAAM) are active metabolites of the opiate l-alpha-acetylmethadol (LAAM), and they contribute to the prolonged actions of the parent compound. Single doses of nor-LAAM, dinor-LAAM, LAAM, methadone and morphine were given intravenously to the chronic spinal dog to determine acute, single-dose effects and their ability to suppress withdrawal in morphine-dependent dogs. These opioids produced dose dependent antinociception, decreases in body temperature and pupillary constriction. For these measures, dinor-LAAM was 1.5 to 3 times and nor-LAAM 6 to 12 times as potent as LAAM. Five hours after the acute administration of LAAM or either of the metabolites, a 1-mg/kg dose of naltrexone given intravenously produced withdrawal, indicating the presence of acute physical dependence. In dogs physically dependent on a daily dose of 125 mg of morphine, nor-LAAM was 9 times as potent as either LAAM or dinor-LAAM in suppressing spontaneous withdrawal 40 hr after the last dose of morphine. The efficacies of LAAM and its demethylated metabolites in the dog for producing acute opiate effects were comparable with those of morphine and methadone. There was a trend, however, for LAAM to suppress the expression of abstinence more fully than either metabolite. The usefulness of LAAM as a treatment for opiate addiction is likely due in part to the equivalent efficacies and higher potencies of its nor and dinor metabolites. PMID- 9353406 TI - Biochemical characterization of the binding of echistatin to integrin alphavbeta3 receptor. AB - Echistatin is a 49-amino-acid peptide belonging to the family of disintegrins that are derived from snake venoms and are potent inhibitors of platelet aggregation and cell adhesion. Integrin alphavbeta3 receptor plays a critical role in several physiological processes such as tumor-induced angiogenesis, tumor cell metastasis, osteoporosis and wound repair. In this study, we have characterized the binding of echistatin to purified integrin alphavbeta3 receptor and the form expressed on human embryonic kidney 293 cells. We show that both purified and membrane-bound integrin alphavbeta3 binds to echistatin with a high affinity, which can be competed efficiently by linear and cyclic peptides containing the RGD sequence. Previous studies have shown that alphavbeta3 binds to vitronectin in a nondissociable manner, whereas an RGD-containing peptide derived from vitronectin binds in a dissociable manner with a Kd of 9.4 x 10(-7) M. Our studies indicate that radiolabeled echistatin binds to alphavbeta3 in a nondissociable manner, similar to native echistatin. However, echistatin does not support the adhesion of 293 cells expressing alphavbeta3 receptor because of poor binding to plastic dishes and is a potent antagonist of the adhesion of these cells to vitronectin. These studies demonstrate that echistatin binding to alphavbeta3 is of high affinity and irreversible similar to vitronectin and provides an alternate ligand for high-throughput screening for alphavbeta3 antagonists. PMID- 9353407 TI - Immunocytochemical localization of the alpha-1B adrenergic receptor and the contribution of this and the other subtypes to vascular smooth muscle contraction: analysis with selective ligands and antisense oligonucleotides. AB - The contribution of the alpha-1B adrenergic receptor (AR) to vascular smooth muscle contraction has been assessed using a combination of immunological, molecular biological and pharmacological approaches. A subtype-selective antibody detected alpha-1B immunoreactivity in the medial layer of the aorta, caudal, femoral, iliac, mesenteric resistance, renal and superior mesenteric arteries. Receptor protection assays and antisense oligonucleotides were used to assess the contribution of the alpha-1B AR to contraction. The alpha-1B AR was implicated in mediating the phenylephrine-induced contraction of the mesenteric resistance artery. The alpha-1D AR was implicated in mediating the contraction of the aorta, femoral, iliac and superior mesenteric arteries. Similarly, the alpha-1A AR was implicated in mediating contraction of the caudal and renal arteries. In vivo application of antisense oligonucleotides targeted to the translational start site of the alpha-1B AR had no effect on the phenylephrine-induced contraction of the femoral or renal arteries. In contrast, antisense oligonucleotides directed against the alpha-1D AR significantly inhibited the phenylephrine response in the femoral artery but had no effect on the renal artery. Application of alpha-1A AR antisense oligonucleotides inhibited the contraction of the renal artery without effect on the femoral artery. These data show that (1) alpha-1B AR immunoreactivity is widely distributed in the same peripheral arteries in which previous studies detected its mRNA, and (2) despite this distribution, receptor protection and antisense oligonucleotide studies indicate that the alpha-1B AR mediates the contraction of only the mesenteric resistance artery. PMID- 9353408 TI - Control of lymphoproliferative and autoimmune disease in MRL-lpr/lpr mice by brequinar sodium: mechanisms of action. AB - Brequinar sodium (BQR) was originally developed as an antitumor drug and subsequently as an immunosuppressant for controlling transplant rejection. It has been widely accepted that the antitumor and immunosuppressive activities of BQR are dependent on its ability to inhibit the enzymatic activity of dihydroorotate dehydrogenase, the fourth enzyme in the de novo pyrimidine synthesis pathway. Recently, we discovered that BQR has the ability to inhibit protein tyrosine phosphorylation in anti-CD3-stimulated murine T lymphocytes and to inhibit the activity of src-related protein tyrosine kinases, p56lck and p59fyn. We examined the in vivo activities of BQR in MRL-lpr/lpr mice. We report that the dose of BQR (10 mg/kg/day) that induced anemia, controlled lymphadenopathy and inhibited autoantibody production, also selectively reduced the pyrimidine nucleotide levels in the bone marrow and in the lymph nodes. Coadministration of uridine (1000 mg/kg/day) with BQR completely normalized pyrimidine nucleotide levels in the bone marrow and lymph nodes, and prevented BQR-induced anemia. However, coadministration of uridine with BQR only partially reversed the anti proliferative effects of BQR, and did not antagonize the inhibitory effect of BQR on autoantibody production. Finally, we report that BQR markedly reduced protein tyrosine phosphorylation in lymph nodes of MRL-lpr/lpr mice. These results collectively suggest that the control of lymphadenopathy and autoantibody production in MRL-lpr/lpr mice by BQR is only partially dependent on inhibition of pyrimidine nucleotide synthesis, and suggest a critical role for in vivo inhibition of protein tyrosine phosphorylation. PMID- 9353409 TI - Design, synthesis and utility of novel benzophenone-containing calcitonin analogs for photoaffinity labeling the calcitonin receptor. AB - Calcitonin (CT) is a 32-amino-acid calciotropic peptide hormone which acts on target cells via a G protein-coupled seven-transmembrane receptor (CTR). In this study, we report the design, synthesis and characterization of four potent bioactive and photoreactive CT analogs, each of which contains a single benzophenone moiety inserted at different and discrete locations within the CT molecule. Replacement of all Lys residues in salmon CT (sCT) with Arg, followed by replacement of hydrophobic residues with a Lys(epsilon-p-benzoylbenzoyl) residue [Lys(epsilon-pBz2)] was found to preserve high biological activity. We substituted Val8, Leu16 and Leu19 by Lys(epsilon-pBz2), and acylated the N terminus by a pBz2 moiety, thus distributing the photoaffinity moiety in the different analogs across a large portion of the CT sequence. With both transfected and endogenous CTRs from several species, all four benzophenone containing analogs were shown to be virtually indistinguishable from the parent sCT analog in both receptor binding properties and stimulation of cAMP accumulation. Upon photolysis, in the presence of CTR, the radioiodinated photoreactive CT analog ([Arg11,18,Lys19(epsilon-pBz2)]sCT (K19)) covalently labels a membrane component of approximately 70 kDa. Receptor cross-linking is inhibited specifically in the presence of excess sCT. We also examined the interaction of these CT analogs with a hemagglutinin (HA) epitope-tagged CTR. The HA-CTR displayed CT binding and CT-dependent cAMP stimulation identical with native CTR. Both K19 and another bioactive analog (-Arg11,18, Lys8(epsilon pBz2)]sCT (K8)) specifically photoaffinity cross-link to the HA-CTR. These benzophenone-containing CT analogs should facilitate studies of hormone-receptor interactions and allow the direct identification of a CT binding domain(s) within the receptor by the analysis of photochemically cross-linked conjugates. PMID- 9353410 TI - Activation of guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase in rabbit aorta by nitroglycerin and sodium nitroprusside. AB - It is generally accepted that cGMP mediates the vascular relaxant effects of nitrovasodilators such as sodium nitroprusside (SNP) and nitroglycerin (NTG). It has been suggested that the relaxant effects of cGMP are mediated via activation of a specific, cGMP-dependent protein kinase (PKG). The objective of this study was to determine whether PKG can be activated by SNP and by NTG in intact strips of rabbit aorta and, if so, whether a good correlation exists between activation of PKG and relaxation of the arteries by the nitrovasodilators. PKG activity was measured by means of a recently described assay using a peptide substrate, BPDEtide, that exhibits good sensitivity and specificity for PKG compared with other protein kinases. Verification of the specificity of the assay for PKG was obtained using MonoQ chromatography to resolve soluble extracts of the rabbit aorta and subsequent immunoblotting to identify the kinase by means of a PKG specific antibody. The role of PKG in vascular relaxation was investigated by simultaneously monitoring the effects of SNP and NTG on cGMP levels, PKG activity ratios and tension in isolated strips of rabbit aorta exposed to varying concentrations of the nitrovasodilators for varying times. The results indicate that PKG can be activated in a concentration- and time-dependent manner by both SNP and NTG in intact vascular preparations and that reasonably good correlations exist between PKG activation and relaxation in these experiments. Although a causal relationship between the two parameters has not been definitely established, these results are consistent with the proposed role for PKG as a mediator of the vascular relaxant effects of cGMP-elevating agents such as SNP and NTG. PMID- 9353411 TI - Activation of guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase in rat vas deferens and distal colon is not accompanied by inhibition of contraction. AB - There is good evidence that in vascular smooth muscle, the relaxant effects of sodium nitroprusside (SNP) are mediated by increases in cGMP levels and activation of cGMP-dependent protein kinase (PKG). However, in rat vas deferens and rat distal colon, cGMP-elevating agents such as SNP and atrial natriuretic factor (ANF) have been shown to elevate cGMP without inducing relaxation. The lack of relaxation might be explained by either lack of activation of PKG by these agents or low levels of PKG in these tissues. The object of the present study was to investigate these possibilities by simultaneously monitoring cGMP levels, PKG activity and contractility in isolated strips of rat vas deferens, rat proximal colon and distal colon exposed to high concentrations of SNP or ANF. Verification of the specificity of the assay for PKG was obtained using MonoQ chromatography to resolve soluble smooth muscle extracts, followed by immunoblotting with a PKG-specific antibody to identify the kinase. In rat vas deferens, 5 mM SNP increased cGMP levels (14-fold) and PKG activity ratios (3.4 fold) but did not inhibit phenylephrine-induced contractions. In both rat proximal and rat distal colon, 100 nM ANF significantly elevated cGMP levels and PKG activity ratios, but only in the proximal colon was inhibition of spontaneous contractions observed. Total PKG activity was much lower (approximately 16 pmol PO4/min/mg protein) in rat vas deferens, which was not relaxed by SNP, than in rabbit aorta (approximately 148 pmol PO4/min/mg), which was relaxed. However, in the rat proximal colon, despite low PKG levels (approximately 11 pmole/min/mg), ANF did inhibit contractions. Thus the inability of the cGMP-elevating agents SNP and ANF to inhibit contractions in rat vas deferens and rat distal colon cannot be explained by either of the possibilities suggested above. PMID- 9353412 TI - Role of Rho protein in lovastatin-induced breakdown of actin cytoskeleton. AB - The Rho GTPases are involved in actin cytoskeleton organization and signal transduction. They need polyisoprenylation for membrane association and activation. Lovastatin, a hydroxymethylglutaryl coenzyme A inhibitor, prevents isoprene synthesis and thereby lipid modification of the Rho protein carboxy terminus. Because lovastatin causes rounding up of cultured cells, we investigated whether the compound acts on the actin cytoskeleton through Rho proteins. Lovastatin treatment decreased F-actin content in a time- and concentration-dependent manner. G-actin content remained unchanged. In lovastatin treated NIH 3T3 cells, the amount of Rho protein which was ADP-ribosylated by Clostridium botulinum exoenzyme C3 decreased in membranes and increased in the cytosol fraction. Cycloheximide prevented lovastatin-induced rounding up of cells. However, after microinjection or direct application of exoenzyme C3, cells treated with cycloheximide and lovastatin rounded up again. On the contrary, lovastatin-treated, round Swiss 3T3 cells reverted to a flat morphology when microinjected with dominant active RhoA (Val14RhoA). Escherichia coli cytotoxic necrotizing factor (CNF1) which activates Rho proteins caused flattening of round, lovastatin-treated NIH 3T3 cells. These results suggest that lovastatin affects the actin cytoskeleton through inactivation of Rho proteins. PMID- 9353413 TI - Carvedilol, a multiple-action neurohumoral antagonist, inhibits mitogen-activated protein kinase and cell cycle progression in vascular smooth muscle cells. AB - Recent findings that the multiple-action neurohumoral antagonist carvedilol inhibits the mitogenic effects of a broad variety of mitogens and produces marked protection against neointima formation after balloon angioplasty injury prompted further study into the molecular and biochemical mechanism of action. In the present study, the effects of carvedilol on mitogen-activated protein (MAP) kinase activity and cell cycle progression were evaluated. Carvedilol produced significant concentration-dependent inhibition of mitogen-induced MAP kinase activity in rat smooth muscle cells. Furthermore, when MAP kinase was purified from mitogen-stimulated cells by FPLC Mono Q chromatography, carvedilol produced direct enzyme inhibition. In the cell-free assay, carvedilol (10 microM) produced 50% inhibition of MAP kinase activity. Cell flow cytometry studies revealed that quiescent rat aortic smooth muscle cells showed 96% of the cell population in the G0/G1 phase of the cell cycle. The addition of serum (10%) increased the number of cells in S and G2/M phases 20% to 40%, respectively. Carvedilol (10 microM) significantly decreased (30-50%) the number of cells in S and G2/M phase. In addition, carvedilol significantly inhibited (>70%) serum-induced stimulation of the S phase-specific marker thymidine kinase. These data suggest that the antimitogenic actions of carvedilol on vascular smooth muscle may be in part due to the inhibition of MAP kinase activity and regulation of cell cycle progression. PMID- 9353414 TI - Protection against septic shock and suppression of tumor necrosis factor alpha and nitric oxide production by dexanabinol (HU-211), a nonpsychotropic cannabinoid. AB - Dexanabinol, HU-211, a synthetic cannabinoid devoid of psychotropic effects, improves neurological outcome in models of brain trauma, ischemia and meningitis. Recently, HU-211 was found to inhibit brain tumor necrosis factor (TNFalpha) production after head injury. In the present study, we demonstrate the ability of HU-211 to suppress TNFalpha production and to rescue mice and rats from endotoxic shock after LPS (Escherichia coli 055:B5) inoculation. In BALB/c mice, a dose of 10 mg/kg LPS, injected i.p., caused 57% and 100% mortality, at 24 and 48 hr, respectively. HU-211, administered i.p. 30 min before lipopolysaccharide (LPS), reduced lethality to 9 and 67% at these time points (P < .05). When coinjected with D-galactoseamine (i.p.), LPS was 100% lethal within 24 hr, whereas eight hourly injections of HU-211 caused mortality of C57BL/6 mice to drop to 10% (P < .001). Administration of LPS to Sprague-Dawley rats resulted in a 30% reduction in the mean arterial blood pressure within 30 min, which persisted for 3 hr. HU 211, given 2 to 3 min before LPS, completely abolished the typical hypotensive response. Furthermore, the drug also markedly suppressed in vitro TNFalpha production and nitric oxide generation (by >90%) by both murine peritoneal macrophages and rat alveolar macrophage cell line exposed to LPS. HU-211 may, therefore, have therapeutic implications in the treatment of TNFalpha-mediated pathologies. PMID- 9353415 TI - Heterologous desensitization of the rat tail artery contraction and inositol phosphate accumulation after in vitro exposure to phenylephrine is mediated by decreased levels of Galphaq and Galphai. AB - Desensitization of alpha-1 adrenoceptor (alpha1AR)-mediated responses in aortic smooth muscle after exposure to catecholamines or alpha1AR agonists has been widely demonstrated. To determine whether exposure to an alpha1AR agonist results in desensitization of alpha1AR-mediated responses in a resistance artery, rat tail artery rings were exposed to 7.5 or 75 microM phenylephrine (PE) for 22 hr in vitro. Norepinephrine-stimulated contraction was significantly reduced in PE exposed tail artery rings. Contractions mediated by the alpha2AR agonists, clonidine and UK 14,304, and by serotonin were also reduced in PE-treated tail artery rings. However, the contractile responses to KCl and ionomycin remained unchanged. Norepinephrine-, PE-, endothelin- and serotonin-stimulated inositol phosphate accumulations were reduced in PE-exposed tail artery rings, whereas KCl and ionomycin-stimulated inositol phosphate accumulation remained unchanged. The density of membrane alpha1ARs, measured by specific [125I]2-([beta-(4 hydroxyphenyl)ethyl]aminomethyl)-1-etralone binding was not changed in PE desensitized tail arteries. Further studies were performed to examine if alterations in receptor/G protein interaction accompanies arterial desensitization. In these studies receptor-stimulated increases in [35S]GTPgammaS binding to G proteins was assessed in membranes obtained from vehicle (control) and PE-treated tail arteries. In control membranes alpha1AR stimulation increased [35S]GTPgammaS binding to Galphaq and Galphai proteins, whereas the alpha2AR agonist UK14,304 activated [35S]GTPgammaS binding to Galphai exclusively. Both PE and UK14, 304-induced responses were reduced in membranes from tail arteries that were exposed to either 7.5 or 75 microM PE for 22 hr. Western blot analyses of G protein alpha and beta subunits demonstrated that Galphaq and Galphai protein levels were decreased in PE-exposed tail artery membranes. These data show that the reduced transmembrane signaling for the alpha1AR in tail artery after in vitro PE exposure is associated with decreases in Galphaq and Galphai protein levels. The reduction in these Galpha proteins also appears to mediate the loss of function of alpha2AR and perhaps of other G protein-coupled receptors. PMID- 9353416 TI - Nor-binaltorphimine precipitates withdrawal and excitatory amino acid release in the locus ceruleus of butorphanol--but not morphine-dependent rats. AB - The relative involvement of kappa opioid receptors in the mediation of behavioral and neurochemical responses to withdrawal from chronic drug treatment with the opioid analgesic butorphanol was studied using in vivo microdialysis to detail extracellular fluid concentrations of glutamate and aspartate within the locus ceruleus. Sprague-Dawley rats were rendered opioid dependent after 3 days of intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/microl/hr) or morphine (26 nmol/microl/hr) and after i.c.v. infusion of saline vehicle (1 microl/hr). Acute withdrawal was precipitated by i.c.v. injection of the selective kappa opioid receptor antagonist nor-binaltorphimine (48 nmol/5 microl) after the 3-day period of infusion. Behavioral signs of withdrawal were detected after nor-binaltorphimine only in butorphanol-dependent rats. Basal levels of glutamate and aspartate were not different between treatment groups. Nor binaltorphimine in the butorphanol-dependent rats increased glutamate to 227% and aspartate to 158% in the initial 15-min sample (P < 0.01). Nor-binaltorphimine did not increase glutamate or aspartate concentrations in the morphine-dependent or saline-treated groups. These results indicate a significantly greater participation of kappa opioid receptors in the development of butorphanol, rather than morphine, dependence and identify a differential neurochemical response to butorphanol withdrawal within a defined brain region, the locus ceruleus. PMID- 9353417 TI - Differential regulation of D2 and D4 dopamine receptor mRNAs in the primate cerebral cortex vs. neostriatum: effects of chronic treatment with typical and atypical antipsychotic drugs. AB - The RNase Protection Assay was used to examine the regulation of D2 and D4 dopamine receptor mRNAs in the cerebral cortex and neostriatum of nonhuman primates after chronic treatment with a wide spectrum of antipsychotic medications (chlorpromazine, clozapine, haloperidol, molindone, olanzapine, pimozide, remoxipride and risperidone). Tiapride, a D2 antagonist that lacks antipsychotic activity, was also included. All drugs were administered orally for 6 months at doses recommended for humans. All antipsychotic drug treatments examined in this study caused a statistically significant up-regulation of both the long and short isoforms of the D2 receptor mRNAs in the prefrontal and temporal cortex. Tiapride, in contrast, significantly up-regulated only the level of D2-long mRNA in these areas. The same drug treatments produced less uniform effects in the neostriatum than in the cortex: clozapine and olanzapine failed to significantly elevate either D2-long or D2-short receptor messages in this structure unlike all other drugs, including tiapride. In both the cerebral cortex and striatum, D4 receptor mRNA was upregulated by certain typical (chlorpromazine and haloperidol) and certain atypical (clozapine, olanzapine and risperidone) antipsychotic agents as well as by tiapride. Other drugs of the typical (molindone and pimozide) and atypical (remoxipride) classes had no effect on D4 mRNA levels in either cortical or striatal tissue. The finding that up-regulation of D2 dopamine receptor mRNAs was a consistently observed effect of a wide range of antipsychotic agents in the cerebral cortex but not in the neostriatum, coupled with the fact that the D2-short isoforms in the cortex were not regulated by a nonantipsychotic D2 antagonist, tiapride, draws attention to the importance of the D2 dopamine receptor in the cerebral cortex as a potentially critical, common site of action of antipsychotic medications. PMID- 9353418 TI - Nitrosylated bovine serum albumin derivatives as pharmacologically active nitric oxide congeners. AB - Although nitrosothiols have been suggested to act as regulators of cell (patho)physiology, little is known about the pharmacology of nitrosylated proteins as nitric oxide (NO.) congeners. We describe the molecular consequences of nitrosylating bovine serum albumin (BSA) at multiple specific sites and demonstrate that the product S-nitrosoproteins exert NO.-like activity. The content of nucleophilic nitrosylation sites (i.e., free sulfhydryl groups) in native BSA was increased by either reduction with dithiothreitol or thiolation with N-acetylhomocysteine. Fourteen moles of nitrogen monoxide (NO)/mol BSA equivalent were then selectively positioned on either the endogenous sulfhydryl groups of reduced BSA or the homocysteine moieties of thiolated BSA, respectively. Each resulting S-nitrosoprotein adduct was an oligomeric mixture across the >2000 kDa to approximately 66 kDa molecular mass range. The BSA derived S-nitrosoproteins were immunoreactive with antibodies against native BSA but evidenced compromised long-chain fatty acid binding. Both types of BSA derived S-nitrosoproteins suppressed human coronary artery smooth muscle cell proliferation to a similar degree (IC50 approximately 70 microM NO. equivalents) and were significantly more effective antiproliferative agents than a standard NO. donor, DETA NONOate. Antiproliferative bioactivity reflected the NO functionalities carried by each protein, but was independent of molecular mass of the nitrosylated BSA adducts. These data exemplify the rational design and characterization of protein-based S-nitrosothiols as NO. congeners and suggest that such agents could have therapeutic potential as NO delivery systems. PMID- 9353419 TI - Inhibition of NFkappaB-mediated interleukin-1beta-stimulated prostaglandin E2 formation by the marine natural product hymenialdisine. AB - Exposure of human rheumatoid synovial fibroblasts (RSF) to interleukin 1beta (IL 1beta) results in the coordinate up-regulation of 85-kDa phospholipase A2 (PLA2) and mitogen-inducible cyclooxygenase (COX II) and subsequent biosynthesis of prostaglandin E2 (PGE2). We have recently demonstrated, through the use of oligonucleotide decoys and antisense, the participation of the proinflammatory transcription factor, nuclear factor kappaB (NFkappaB), in the regulation of the prostanoid-metabolizing enzymes. Hymenialdisine, a marine natural product has recently been characterized as an inhibitor of NFkappaB activation and exposure of IL-1-stimulated RSF-inhibited PGE2 production in a concentration-dependent manner (IC50 approximately 1 microM). Alternatively, both an analog, aldisine, and the protein kinase C inhibitor, RO 32-0432, were without affect. Direct action of hymenialdisine on IL-1-induced NFkappaB activation was demonstrated by a significant reduction (approximately 80%) in NFkappaB binding to the classical kappaB consensus motif (as assessed by electrophoretic mobility shift assay) and inhibition of stimulated p65 migration from the cytosol of treated cells (as assessed by Western analysis). Consistent with the role of NFkappaB in the transcriptional regulation of COX II and 85-kDa PLA2, hymenialdisine-treated RSF did not transcribe the respective mRNAs in response to IL-1. This led to reductions in their respective protein levels and subsequent reductions in the ability to produce PGE2. Specificity of action is suggested as IL-1-stimulated interleukin-8 (IL-8) production, which is known to be an NFkappaB-regulated event, was also inhibited by hymenialdisine, whereas IL-1-induced production of vascular endothelial growth factor, a non-NFkappaB-regulated gene, was not affected by exposure to hymenialdisine. Taken together, hymenialdisine inhibits IL-1-stimulated-RSF PGE2 formation acting predominately through modulation of NFkappaB activation and offers an interesting novel tool to evaluate the role of NFkappaB in inflammatory disease. PMID- 9353420 TI - Activation of metabotropic glutamate receptors in the rat nucleus accumbens increases locomotor activity in a dopamine-dependent manner. AB - The effect on locomotor activity of in vivo activation of metabotropic glutamate receptors (mGluRs) in the nucleus accumbens (NAcc) was investigated in rats. Bilateral intracranial microinjections into the NAcc of the selective mGluR agonist, 1-aminocyclopentane-trans-1,3-dicarboxylic acid [(1S,3R)-ACPD], were made in the freely moving rat and locomotor activity was subsequently measured for 2 hr. Different groups of rats injected with one of four doses of (1S,3R) ACPD (0.005, 0.05, 0.5, or 2.5 nmol/0.5 microl/side) showed significant dose dependent increases in both horizontal and vertical locomotor activity relative to control rats that received injections of the saline vehicle. Time-course analyses revealed that these effects, in a manner similar to the locomotor hyperactivity produced by the injection of amphetamine into the NAcc, were most pronounced in the initial 30 min after injection and no longer present after 1 hr of testing. These locomotor-activating effects of (1S,3R)-ACPD were blocked by the co-injection of the mGluR antagonist, (RS)-alpha-methyl-4 carboxyphenylglycine (2.5 nmol/side), as well as of the dopamine receptor antagonist, fluphenazine (2.0 or 9.8 nmol/side), which suggests that they depend on dopamine neurotransmission. These findings indicate that mGluRs play an important role in the production of locomotor behaviors involving DA-excitatory amino acid interactions in the NAcc. PMID- 9353421 TI - Carbohydrate vaccines that induce antibodies against cancer. 1. Rationale. PMID- 9353422 TI - Carbohydrate vaccines that induce antibodies against cancer. 2. Previous experience and future plans. PMID- 9353423 TI - Taxol-mediated changes in fibrosarcoma-induced immune cell function: modulation of antitumor activities. AB - The anticancer drug taxol (paclitaxel) inhibits tumors through multiple cytotoxic and cytostatic mechanisms. Independently of these mechanisms, taxol induces distinct immunological efficacy when it acts as a second signal for activation of tumoricidal activity by interferon gamma (IFN gamma)-primed murine normal host macrophages. We reported that tumor-distal macrophages, which mediate immunosuppression through dysregulated nitric oxide (NO) and tumor necrosis factor alpha (TNF alpha) production, are differentially regulated by taxol. Because taxol influences tumor cell growth dynamics and activates immune cell populations, we assessed the ex vivo immunosuppressive and antitumor activities of taxol-treated normal host and tumor-bearing host (TBH) macrophages. Pretreatment of such cells with taxol partly reconstituted T cell alloantigen reactivity, suggesting that taxol mediates a limited reversal of TBH macrophage immunosuppressive activity. Taxol-treated TBH macrophages significantly suppressed the growth of fibrosarcoma cells (Meth-KDE) through soluble effector molecules and promoted direct cell-mediated cytotoxicity, indicating that taxol enhanced tumor-induced macrophage antitumor activities. Tumor-induced helper T cells, however, showed a higher sensitivity to direct taxol-induced suppression. These data demonstrate that taxol exerts pleiotropic effects on antitumor immune responses with the capacity to abate the immunosuppressive activities of macrophages and promote macrophage-mediated antitumor activities simultaneously, but also directly modulating T cell reactivity. Collectively, these studies suggest that the antineoplastic drug taxol may impart antitumor activity through an immunotherapeutic capacity. PMID- 9353424 TI - Augmentation of impaired tumoricidal function in alveolar macrophages from lung cancer patients by cocultivation with allogeneic, but not autologous lymphocytes. AB - It has been reported that the in vitro development of tumoricidal function in alveolar macrophages from lung cancer patients is reduced significantly when compared to that in peripheral blood monocytes from the same patients or alveolar macrophages from control patients. In the present investigation, a method for potentiating the development of tumoricidal function in alveolar macrophages from lung cancer patients is described. This method, which relies on priming the macrophages with purified, allogeneic peripheral blood lymphocytes from normal donors, could not be demonstrated when autologous lymphocytes from lung cancer patients were used in the priming coculture. The augmentation of tumoricidal function appears to be mediated by one or more soluble factors, since supernatants from cocultures of alveolar macrophages and allogeneic peripheral blood lymphocytes could enhance the cytotoxic function of freshly obtained alveolar macrophages. Furthermore, it appears that NK cells are necessary for this effect, since depletion of CD56+/CD57+ cells from allogeneic lymphocytes eliminated their capacity to enhance alveolar macrophage cytotoxic function. The augmentation of cytotoxic function elicited in alveolar macrophages by this method was not associated with changes in the secretion of tumor necrosis factor alpha, or interleukin 1 beta. PMID- 9353425 TI - Modulation of tumoricidal function in alveolar macrophages from lung cancer patients by interleukin-6. AB - Previous studies have demonstrated that alveolar macrophages from lung cancer patients are impaired in their ability to develop tumoricidal function when stimulated by activators such as interferon gamma + lipopolysaccharide. However, these same macrophages have been shown to develop significant tumoricidal function when precultured with macrophage-depleted allogeneic peripheral blood lymphocytes from normal donors, an effect that was lost by the elimination of natural killer cells from the allogeneic lymphocyte population. In the present study, the effect of each activation condition on the expression of mRNA for interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF alpha) and IL-6 was determined using reverse transcription/polymerase chain reaction. The results show that the non-permissive activation condition is associated with the expression of mRNA for IL-6 while the permissive activation condition is not. Antibodies against IL-6 were subsequently shown to permit the development of tumoricidal function in alveolar macrophages stimulated with interferon gamma + lipopolysaccharide while IL-6 protein was shown to inhibit the stimulatory action of allogeneic lymphocytes on the development of tumoricidal function in the same alveolar macrophages. Neither the permissive (i.e. allogeneic lymphocyte stimulation) nor the non-permissive (i.e. interferon gamma + lipopolysaccharide) activation condition had any effect on the capacity of alveolar macrophages from lung cancer patients to express mRNA for IL-1 alpha, IL 1 beta or TNF alpha. These results show that IL-6 can regulate the ability of alveolar macrophages from lung cancer patients to be stimulated by interferon gamma + lipopolysaccharide to develop significant tumoricidal function. They also show that allogeneic lymphocytes have the capacity to down-regulate IL-6 mRNA synthesis by alveolar macrophages thereby permitting the development and/or expression of macrophage tumoricidal function. PMID- 9353426 TI - Interleukin-13 secretion by normal and posttransplant T lymphocytes; in vitro studies of cellular immune responses in the presence of acute leukaemia blast cells. AB - T lymphocyte secretion of interleukin-13 (IL-13) in response to different activation signals was characterized in vitro. IL-13 release was investigated when virus transformed B lymphocytes or acute myelogenous leukaemia (AML) blasts were used as accessory cells during T cell activation. First, a majority of both CD4+ and CD8+ TCR alpha beta + T lymphocyte clones, derived from normal individuals and bone marrow transplant recipients, secreted IL-13 in response to a standardized mitogenic activation signal (phytohaemagglutinin + IL-2 + B lymphocyte accessory cells). The CD4+ cells showed significantly higher IL-13 levels than the CD8+ subsets. Second, when leukaemic accessory cells (more than 95% AML blasts) were used during T cell activation, IL-13 was released both during alloactivation of normal T lymphocytes and during mitogen activation of posttransplant T cells. Third, when normal T lymphocytes were stimulated with allogeneic AML blasts, addition of IL-13-neutralizing monoclonal antibodies decreased interferon gamma levels. Although addition of IL-13-neutralizing antibodies did not alter granulocyte-colony-stimulating factor secretion by allostimulating AML blasts, altered blast proliferation was detected for certain patients. Thus, most T cell clones can release IL-13, and IL-13 can modulate cytokine responses during T cell recognition of allogeneic AML cells. PMID- 9353427 TI - HLA-A2 antigen status predicts metastasis and response to immunotherapy in gastric cancer. AB - Our previous studies have shown that HLA-DR4 and -B52 antigens are associated with an increased risk of lymph node metastasis in patients with gastric cancer. We hypothesized that a putative HLA antigen, correlated with a low risk of lymph node metastasis, may also be correlated with the response to anticancer therapy. The microcytotoxicity assay was used to examine 49 HLA antigens of the A, B, C, DR, and DQ loci, and the association between HLA class I and II antigen status and lymph node metastasis in 847 patients with gastric cancer as well as the response to the therapy in 739 patients were analyzed. HLA-A2 antigen was significantly associated with a low risk of lymph node metastasis in patients with T2-T4 advanced cancer [58.8% compared to 37.0% in patients with lymph node metastasis; corrected P, Pc (98), = 0.011], especially in those with moderately differentiated adenocarcinoma [71.0% compared to 26.4% in patients with lymph node metastasis, Pc (294) = 0.00294] and with a better response to post-operative immunotherapy using protein-bound polysaccharide K (PSK), a nonspecific immunomodulator, than to chemotherapy. HLA alleles may be associated with resistance or susceptibility to lymph node metastasis and HLA-A2 antigen may be a useful predictor of the response to PSK. The data suggest that the predictive power of this HLA antigen may prove useful in the selection of anticancer therapy. PMID- 9353428 TI - Radiation therapy for early glottic carcinoma (T1N0M0). The adverse effect of treatment interruption. AB - PURPOSE: Clarification of the adverse effects of treatment interruption on the local control of early glottic carcinoma. PATIENTS AND METHODS: From May 1982 through February 1992, 273 patients with early glottic carcinoma (T1N0M0) were treated at this department. Of 253 patients administered 60 Gy in 30 fractions 77 patients had no treatment interruption and treatment was completed within 6 weeks (group I), overall treatment time was prolonged for 176 patients: 141 patients 43 to 49 days (group II) and 35 patients 50 to 62 days (group III). Treatment was interrupted due to public holidays (156 cases), patients convenience (13 cases) and severe mucosal reactions (seven cases). The major reason was public holidays, 91% in group II and 80% in group III. RESULTS: The 3-year recurrence-free survival rates were for group I 95%, group II 89% and group II 80%. Survivals for groups I and II, groups II and III and groups I and III were essentially the same. At 40 Gy tumor clearance was more than 50% in the 3-groups. For complete clearance cases at 40 Gy, recurrence-free survival was essentially the same for the 3 groups although for incomplete clearance cases, statistically significant difference for groups I and III (log-rank test p = 0.0004; Wilcoxon test p = 0.0004) and marginally significant difference for groups II and III (p = 0.0157, p = 0.0045) but no difference for groups I and II (p = 0.0669, p = 0.0853) were noted by adjusting the p-value. CONCLUSION: Prolongation of overall treatment time and tumor clearance at 40 Gy appeared to be a factor of the local control. PMID- 9353429 TI - Dosimetric aspects of physical and dynamic wedge of Clinac 2100C linear accelerator. AB - AIM: To investigate variation of wedge factors on field size and depth for physical and dynamic wedges of identical wedge angles for Clinac 2100C linear accelerator and its clinical implementation. MATERIAL AND METHODS: A computer controlled water phantom dosimetric system is used to generate profile data for physical wedges, whereas a 0.6 cm3 ion chamber is used for generation of profiles for dynamic wedge and wedge factors for both types of wedges. The method has been discussed to handle the dynamic wedge dosimetry in absence of linear array of detectors or film densitometer. RESULTS: A systematic dependence on wedge factor is observed for physical wedge, with respect to depth and wedge angle but not depending on field size. Whereas dynamic wedge shows strong dependence on field size and is not systematic because the dynamic wedge is controlled by segmented treatment tables depending on field size and energy and no significant variation is observed on depth for various wedge angles. The handling of beam data in a commercially available treatment planning system is discussed and a comparison has been made for iso-doses of both types of wedges. CONCLUSION: The dynamic wedge isodose curves shows rather straight lines than physical wedge but larger hot spots at thin edge which needs careful consideration during planning. PMID- 9353430 TI - Star Wars. PMID- 9353431 TI - Consequences of being accused of malpractice. PMID- 9353432 TI - Resident evaluations: a computerized approach. AB - OBJECTIVE: Accurate and timely evaluation of resident performance is an essential part of a high-quality residency training program. To augment a periodic attending staff round-table discussion of each resident, a computerized resident evaluation software program has been used at our institution. CONCLUSION: Resident grading software not only provides more efficient data collection but also minimizes group influence bias and provides a wealth of statistical data on both individual residents and program sections. Data entry can be performed on a Windows NT networked IBM-compatible computer in each staff members' office at any convenient time. A previously time-consuming task has been transformed into a sophisticated, quick evaluation process with greater reliability, more meaningful and quantifiable data, and more simplified reporting mechanisms. PMID- 9353433 TI - Interactive virtual endoscopy. PMID- 9353435 TI - Decreased risk of subsequent colonic cancer in patients undergoing polypectomy after barium enema: analysis based on data from the preendoscopic era. AB - OBJECTIVE: Published data from the Mayo Clinic gathered during the preendoscopic era were analyzed to show that the risk of subsequent colonic carcinoma is reduced in patients with benign-appearing polyps that are revealed by radiology who then undergo polypectomy. MATERIALS AND METHODS: Data from the Mayo Clinic gathered during a 6-year period before the availability of endoscopy were used to determine the effect on the subsequent risk of colonic carcinoma if the benign appearing polyps initially revealed by radiology had been removed rather than left in place and followed up by serial barium enemas. Data were from 226 patients with benign-appearing polypoid lesions of the colon that were 1 cm in diameter or larger and had been followed up by periodic barium enemas. The period of radiologic surveillance was 12-229 months (mean, 68 months). Between two and 17 barium enemas (mean, 5.2) were performed on each patient. The clinical follow up period was 12-242 months (mean, 140 months). RESULTS: Twenty-one adenocarcinomas developed at the site of the index polypoid lesion as found on follow-up barium enema examinations of these patients. Eleven additional adenocarcinomas of the colon were found at sites remote from that of the index lesion. If the index polyp had been removed when initially diagnosed radiologically, 66% fewer subsequent carcinomas would have occurred in these patients during the average of 11 years of clinical follow-up. CONCLUSION: Excision of benign-appearing polyps found on initial barium enema examinations would result in a significant decrease in the subsequent risk of colonic adenocarcinoma. PMID- 9353434 TI - CT colonoscopy of colorectal neoplasms: two-dimensional and three-dimensional virtual-reality techniques with colonoscopic correlation. AB - OBJECTIVE: The aim of this study was to compare the diagnostic accuracy of two dimensional (2D) CT colonography and three-dimensional (3D) virtual colonoscopy with conventional colonoscopy in patients who have suspected colorectal neoplasms. SUBJECTS AND METHODS: Twenty patients were studied (eight women and 12 men; mean age, 53 years; range, 42-85 years). All patients had findings on conventional colonoscopy suggestive of colorectal carcinoma and underwent colonic CT within 3 hr of endoscopy. Two-dimensional CT colonography and 3D virtual colonoscopy images were generated from the same data set that was obtained from thin-section helical CT of the abdomen and pelvis after rectal insufflation of room air. Three-dimensional virtual colonoscopy images were obtained by downloading CT data to a workstation equipped with commercially available software. Volume- and perspective-rendering techniques were used to achieve interactive, 3D virtual "fly-through" examinations of the colonic mucosa. The results of 2D CT colonography and 3D virtual colonoscopy were compared with the findings of conventional colonoscopy and correlated with surgical and pathologic outcome where possible. RESULTS: Twenty masses (defined as intraluminal projections 2 cm or larger in diameter) and 15 polyps (defined as projections smaller than 2 cm in diameter) were identified in our study group. All masses and 14 of 15 polyps were successfully shown on 2D colonography. Three findings of polyps on 2D colonography were false-positive, and one was false-negative. Three dimensional virtual colonoscopy revealed 19 of 20 masses and 13 of 15 polyps. On conventional colonoscopy, all 20 masses and 13 of 15 polyps were identified, with one false-positive finding of a malignant stricture in a normal colon. Complete examination of the colon was possible in 18 of 20 patients using the 2D technique and in 17 of 20 patients using 3D virtual colonoscopy, whereas conventional colonoscopy showed the entire colon in only 12 of 20 patients. CONCLUSION: Two dimensional CT colonography and 3D virtual colonoscopy are complementary and effective techniques for examining the colon in patients with suspected colorectal carcinoma. CT techniques offer several advantages over conventional colonoscopy including the ability to detect abnormalities proximal to obstructing carcinomas, accurate localization of abnormalities within the colon, and good patient tolerance. These CT techniques may play an important role in future diagnosis of colorectal cancer and for screening patients at risk. PMID- 9353436 TI - Transrectal sonography in staging rectal carcinoma: the role of gray-scale, color flow, and Doppler imaging analysis. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of combining gray-scale sonography with color-flow imaging and pulsed Doppler transrectal sonography in the staging of rectal carcinoma. SUBJECTS AND METHODS: Thirty-nine patients with primary rectal carcinoma underwent transrectal sonography. The rectal masses were staged T1-T2 or T3-T4 on the basis of gray-scale imaging. The local nodes were classified as benign or malignant on the basis of size and echogenicity. In 22 patients, color-flow imaging and pulsed Doppler imaging of the rectal mass and of the local lymph nodes were performed. The peak systolic velocity (PSV) and end diastolic velocity were documented, and the resistive index was calculated. RESULTS: Gray-scale imaging alone was used to stage T1-T2 masses with 88% sensitivity and 82% specificity. T3-T4 masses were staged with 82% sensitivity and 88% specificity. Overall accuracy was 85%. Gray-scale imaging of lymph nodes using a discriminatory size of less than or equal to 5 mm for benign nodes and greater than 5 mm for malignant nodes yielded a sensitivity of 100%, a specificity of 28%, and an accuracy of 52%. Using receiver operating characteristic curve analysis, we determined that a size of greater than or equal to 7 mm was optimal for characterizing nodes. Such a size provided an accuracy of 83%. PSV of less than 25 cm/sec distinguished T3-T4 from T1-T2 rectal masses with 75% sensitivity, 80% specificity, and 77% accuracy. A PSV of greater than 20 cm/sec classified a node as malignant with 100% sensitivity, 62% specificity, and 76% accuracy. A resistive index of greater than 0.61 classified a node as malignant with 71% sensitivity, 85% specificity, and 80% accuracy. CONCLUSION: Color-flow imaging and pulsed Doppler imaging are useful additions to gray-scale transrectal sonography in staging primary rectal carcinomas. The combination has most value when evaluating perirectal nodes. PMID- 9353437 TI - Pericolic mesenteric lymph nodes: an aid in distinguishing diverticulitis from cancer of the colon. AB - OBJECTIVE: This study was done to determine if the detection of pericolic lymph nodes on CT scans could be used to differentiate cancer of the colon from diverticulitis. MATERIALS AND METHODS: We retrospectively evaluated 58 CT scans from 57 patients with proven diverticulitis or cancer of the colon. The CT scans were evaluated by five board-certified radiologists who were unaware of the proven diagnosis. Consensus opinions regarding the presence and size of pericolic lymph nodes were recorded. These data were correlated with the proven diagnoses to determine the correlation between the observed findings and the type of colonic abnormality. Fisher's exact test was used to determine statistical significance. RESULTS: Lymph nodes were seen in 22 (71%) of 31 cases of colonic cancer and in four (15%) of 27 cases of diverticulitis. The lymph nodes were 0.5 2.5 cm in short-axis diameter. We saw no difference in node size for patients with colonic cancer versus patients with diverticulitis. The nodes were most commonly located along the blood vessels in the mesenteric fat. Statistical analysis showed a significant difference (p < .001) in the frequency but not in the size of nodes between the two groups of patients. The detection of nodes resulted in a diagnostic sensitivity and specificity for colonic cancer of 71% and 85%, respectively. CONCLUSION: Pericolic lymph nodes are seen much more frequently in patients with colonic cancer than in patients with diverticulitis. The detection of pericolic lymph nodes in patients suspected of having diverticulitis should raise the suspicion of underlying colonic cancer that should, in turn, prompt additional evaluation. PMID- 9353438 TI - Laparoscopic sonography of peripancreatic tumors: preliminary experience. AB - OBJECTIVE: This study was performed to evaluate the use of laparoscopic sonography in patients with suspected peripancreatic tumors and to assess the impact of laparoscopic sonography on patient management. SUBJECTS AND METHODS: In a prospective study, 24 patients with suspected pancreatic malignancy underwent CT, laparoscopy, and laparoscopic sonography. The pancreas, peripancreatic vasculature, liver, and porta hepatis were evaluated in each patient. Metastases (hepatic, peritoneal, or nodal), extrapancreatic extension of tumor, or vascular encasement was considered evidence of unresectable disease. Histopathology was the standard of reference; unresectable disease was confirmed by biopsy. At the completion of the laparoscopic sonography, each examination was scored according to impact analysis categories that had been prospectively established. RESULTS: Peripancreatic vasculature was adequately shown by laparoscopic sonography in 22 patients (92%), of whom 12 patients had histopathologic evidence of vascular encasement. All 12 cases of vascular encasement were revealed by laparoscopic sonography, and 10 of 12 cases of vascular encasement were revealed by CT. Liver lesions were seen in eight patients (33%). One hemangioma was shown solely by laparoscopic sonography; the other seven liver lesions were revealed by CT, laparoscopy, or both. In six patients (25%), laparoscopic sonography was used to guide biopsy of lesions that were not seen by laparoscopy. Impact analysis showed that laparoscopic sonography provided additional information in eight patients (33%) and altered management in four patients (17%). Of those patients for whom laparoscopic sonography altered management, three patients underwent successful resection after laparoscopic sonography two of these patients had suspected vascular encasement on CT but laparoscopic sonography revealed normal vessels, and the third patient had CT evidence of a liver lesion that was shown to be a cyst on laparoscopic sonography. A fourth patient was spared laparotomy when laparoscopic sonography revealed unsuspected vascular encasement. CONCLUSION: Preliminary experience suggests that laparoscopic sonography may aid diagnosis and alter management in patients with suspected pancreatic neoplasms. PMID- 9353439 TI - Inflammatory pancreatic masses: differentiation from ductal carcinomas with contrast-enhanced sonography using carbon dioxide microbubbles. AB - OBJECTIVE: The aim of this study was to evaluate the clinical efficacy of contrast-enhanced sonography using carbon dioxide microbubbles to differentiate inflammatory pancreatic masses from ductal carcinomas of the pancreas. SUBJECTS AND METHODS: Fifty-five patients, including 35 patients with ductal carcinomas and 20 with inflammatory pancreatic masses, underwent contrast-enhanced sonography, CT, and digital subtraction angiography (DSA). Carbon dioxide microbubbles were prepared by mixing 10 ml of carbon dioxide and the same amount of 25% soybean oil vigorously. Carbon dioxide microbubbles were injected through an angiographic catheter that was placed in the celiac axis. Vascularity of the tumors as determined by those three techniques was interpreted by three physicians who had no knowledge of the pathologic results. RESULTS: Contrast enhanced sonography was best at revealing tumor vascularity among the three techniques. On contrast-enhanced sonography, 19 (95%) of the 20 inflammatory pancreatic masses were isovascular and 32 (91%) of the 35 ductal carcinomas were hypovascular. In contrast, the isovascularity of inflammatory masses was five (25%) on CT, and two (10%) on DSA, respectively. The sensitivity and accuracy rate of differentiating both diseases on contrast-enhanced sonography were 98% and 95%, respectively; on CT, they were both 73%; and on DSA they were both 67%. From our results, an isovascular mass is probably an inflammatory mass, whereas a hypovascular mass is most likely a ductal carcinoma on contrast-enhanced sonography. CONCLUSION: Contrast-enhanced sonography can help differentiate an inflammatory pancreatic mass from a ductal carcinoma. PMID- 9353440 TI - Venous thrombosis of pancreatic transplants: diagnosis by duplex sonography. AB - OBJECTIVE: Our objective was to determine whether elevated pancreatic transplant arterial resistive index (RI) and absence of venous flow correlate with pancreatic transplant venous thrombosis. MATERIALS AND METHODS: Thirteen episodes of surgically documented pancreatic venous thrombosis occurred in 175 pancreases that had been transplanted over a 3-year period. Duplex sonography was performed before surgical exploration in 11 cases. We retrospectively reviewed these 11 sonograms to determine whether blood was flowing in the veins and arteries of the graft. The RI was calculated from all pancreatic artery waveforms. We compared these arterial RIs and the presence or absence of venous flow with those of pancreatic grafts without venous thrombosis to determine sensitivity and specificity. RESULTS: In the venous thrombosis group, thrombosis occurred within 12 days of transplantation (mean, 3.5 days) in all 11 cases. Six cases of thrombosis (55%) occurred within 1 day. Arterial flow was detected within the graft in nine cases (82%) and in the stump of the donor artery between the graft and the recipient iliac artery in the two remaining cases. Antegrade diastolic flow was absent in all arterial tracings. Diastolic flow reversal was present in seven (78%) of nine grafts with detectable intrapancreatic arterial flow. Arterial RIs ranged from 1.00 to 2.00 (mean +/- SD, 1.27 +/- 0.29). Intrapancreatic venous flow was absent in all 11 cases. In the control group (43 examinations in 34 patients) RIs ranged from 0.46 to 1.29 (mean +/- SD, 0.72 +/- 0.18). Two of 43 arterial tracings had diastolic flow reversal (RI > 1.0). Venous flow was present in all examinations in the control group. A statistically significant difference existed between the RIs in the thrombosis group and the RIs in the control group (p = .0001). CONCLUSION: Reversal of diastolic flow in pancreatic transplant arteries is highly specific for detection of graft venous thrombosis during the first 12 days after transplantation. Our findings suggest that an RI greater than or equal to 1.00 and absence of venous flow, in combination, are highly sensitive and specific for the diagnosis of pancreatic graft venous thrombosis. PMID- 9353442 TI - Using uncovered metallic endoprostheses to treat recurrent benign esophageal strictures. AB - OBJECTIVE: The management of recurrent benign esophageal strictures is a difficult clinical problem, especially in patients who are not surgical candidates. We evaluated the role of uncovered metallic endoprostheses in four patients who had strictures that were resistant to repeated balloon dilatation. CONCLUSION: Our preliminary experience indicates that uncovered metallic endoprostheses can be effective in treating a select group of patients who have benign esophageal strictures and for whom multiple dilatations have failed. It is also important to note that epithelial hyperplasia can result in stenoses and recurrent dysphagia. PMID- 9353441 TI - Helical CT combined with contrast material administered only through the colon for imaging of suspected appendicitis. AB - OBJECTIVE: Helical CT combined with contrast material administered by mouth and through the colon has been shown to be accurate for appendiceal imaging. This investigation was performed to determine if helical CT combined with contrast material administered only through the colon has comparable accuracy. SUBJECTS AND METHODS: One hundred patients prospectively underwent appendiceal CT imaging with thin-collimation, helical scanning limited to the lower abdomen and upper pelvis after contrast material was administered only through the colon. CT results were correlated with surgical and pathologic findings at appendectomy (56 patients), other surgery (three patients), or clinical follow-up at least 2 months after the CT scan (41 patients). RESULTS: Fifty-three CT scans were interpreted as positive for appendicitis, including 52 true-positives (with surgical-pathologic correlation) and on false-positive (with clinical follow-up). Forty-seven CT scans were interpreted as negative for appendicitis, including 40 true-negatives with clinical follow-up, three true-negatives with appendectomy and pathologic correlation, three true-negatives with other surgery and pathologic correlation, and one false-negative with appendectomy and pathologic correlation. CT had a 98% sensitivity, 98% specificity, 98% positive predictive value, 98% negative predictive value, and 98% accuracy for diagnosing or excluding appendicitis. In 47 normal appendix cases at CT, the appendix was seen in 44 cases (94%), and an alternative diagnosis was identified in 29 cases (62%). CONCLUSION: For diagnosing appendicitis, helical CT combined with contrast material administered only through the colon proved to be as accurate (98%) as helical CT combined with contrast material administered by mouth and through the colon. Helical CT with contrast material administered only through the colon also could be performed immediately and without any of the potential patient risks or discomforts of contrast material administered i.v. or by mouth. PMID- 9353443 TI - Treatment of colonic obstructions with metallic stents: indications, technique, and complications. PMID- 9353444 TI - Single-shot T2-weighted MR imaging of the upper abdomen: preliminary experience with double-echo HASTE technique. PMID- 9353445 TI - MR cholangiopancreatography: techniques and clinical applications. PMID- 9353446 TI - Breath-hold MR cholangiopancreatography using a HASTE sequence: comparison of single-slice and multislice acquisition techniques. PMID- 9353449 TI - Cross-sectional imaging findings in a case of polyarteritis nodosa with a ruptured hepatic artery aneurysm. PMID- 9353448 TI - Hepatosplenic fungal disease: diagnostic accuracy and spectrum of appearances on MR imaging. AB - OBJECTIVE: We describe our 6-year experience in the prospective examination of patients with suspected hepatosplenic fungal disease to show the diagnostic accuracy of MR imaging and the spectrum of appearances on MR images. SUBJECTS AND METHODS: All patients who underwent MR examination for suspected hepatosplenic fungal disease from January 1990 to January 1997 in three university institutions were included in the study. Patients presented with persistent fever or no response to antibacterial antibiotics. Patients were grouped as acute, subacute treated, and chronic treated, according to the duration of their symptoms. Patients with 2 weeks or fewer of possible infection were acute presentation, patients on antifungal therapy longer than 2 weeks but shorter than 3 months were subacute treated presentation, and patients on antifungal therapy for 3 months or longer or who had completed antifungal therapy and had a history of hepatosplenic fungal disease were chronic treated presentation. MR studies were prospectively interpreted for the presence of hepatosplenic fungal lesions. The appearances of fungal lesions in patients in each category were determined. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for detecting lesions in patients with acute presentation were also determined. RESULTS: Sixty-nine patients were included in the study. MR imaging revealed signs consistent with hepatosplenic fungal lesions in 22 (32%) of 69 patients and no demonstration of hepatosplenic lesions in 47 (68%) of 69 patients. In the 60 patients with acute presentation, MR imaging in 13 patients revealed hepatosplenic lesions that were interpreted as fungal disease. True-positive lesions were present in 11 of these 13 patients. These lesions measured less than 1 cm in diameter and were best shown as well-defined high-signal-intensity foci on T2-weighted images. The remaining two of the 13 patients had false-positive lesions; one was shown to have tuberculosis, and the other had graft-versus-host disease. For acute presentation, MR sensitivity was 100%, specificity was 96%, positive predictive value was 85%, negative predictive value was 100%, and accuracy was 97%. In the five patients with subacute presentation, lesions were present that measured less than 1 cm in diameter and were best shown as mildly hyperintense on T1-weighted images. A perilesional ring nearly void of signal intensity was seen on unenhanced and gadolinium-enhanced T1-weighted images in all five patients. The four patients with chronic healed lesions all had lesions that were 1-3 cm in diameter with irregular, angular polygonal margins. These lesions, which were best shown on images obtained immediately after gadolinium administration, appeared as regions of diminished enhancement with no perilesional changes. CONCLUSION: MR imaging has high diagnostic accuracy for the diagnosis of acute hepatosplenic fungal disease. Patients with acute, subacute treated, and chronic healed presentations may have lesions that can be distinguished by their MR appearances. PMID- 9353447 TI - Use of carbon dioxide microbubble-enhanced sonographic angiography for transcatheter arterial chemoembolization of hepatocellular carcinoma. AB - OBJECTIVE: Our objective was to determine the usefulness of sonographic angiography with carbon dioxide microbubbles during transcatheter arterial chemoembolization for hepatocellular carcinoma. SUBJECTS AND METHODS: Thirty-four patients with hepatocellular carcinoma underwent sonographic angiography during transcatheter arterial chemoembolization. Digital subtraction angiography failed to reveal tumors in 27 patients. Tumor stain became obscure on digital subtraction angiography after the catheter was inserted into distal branches in seven patients. Sonographic angiography was performed after injection of carbon dioxide microbubbles into the hepatic artery selected for transcatheter arterial chemoembolization. RESULTS: In angiographically undetectable hepatocellular carcinomas, sonographic angiography revealed tumor vascularity in 17 patients in whom transcatheter arterial chemoembolization was then performed. For the two patients who underwent a second transcatheter arterial chemoembolization, the existence of alternative feeding vessels was confirmed by sonographic angiography. In the remaining 10 patients, tumor vascularity was not seen on sonographic angiography; percutaneous ethanol injection therapy was then performed. Sonographic angiography clearly revealed tumor vascularity in patients in whom staining became obscure on digital subtraction angiography after the catheter was inserted into a peripheral branch of the hepatic artery. In all 24 patients who underwent transcatheter arterial chemoembolization, sonographic angiography was useful for determining the artery suitable for transcatheter arterial chemoembolization and for monitoring tumor perfusion in the selected artery. CONCLUSION: Sonographic angiography can be used to determine not only the therapeutic strategy for treatment of hepatocellular carcinoma but also whether the tumor is supplied by the artery selected for transcatheter arterial chemoembolization, especially when the tumor is not revealed by digital subtraction angiography. PMID- 9353450 TI - Imaging of fallopian tube tumors. AB - OBJECTIVE: The purposes of this study were to investigate the imaging findings in patients with primary fallopian tube neoplasms and to determine whether specific imaging features favor the preoperative diagnosis of fallopian tube tumors (FTT). MATERIALS AND METHODS: Computerized search of medical records from 1984 to 1994 identified 20 patients with a discharge diagnosis of primary fallopian tube carcinoma. Medical records, imaging studies, and pathology findings were reviewed. Eleven patients had available preoperative imaging. RESULTS: Seventeen of 20 patients with primary FTT had unilateral disease. Of these 17, preoperative imaging was available in nine, showing four solid adnexal masses, four complex cystic adnexal masses, and one normal adnexa. The preoperative imaging of these nine patients included six sonographic and five CT studies. Three patients with primary FTT had bilateral tumors, and preoperative imaging was available for two patients: Two sonographic studies and one CT study showed one complex cystic adnexal mass and three normal adnexa. CONCLUSION: Primary FTT commonly presents as an adnexal mass on preoperative imaging and mimics other pelvic malignancies, especially ovarian carcinoma. Making a specific preoperative diagnosis is difficult; however, because primary FTT is unlikely to be confused with a benign process, delay in diagnosis is rare. PMID- 9353452 TI - Three-dimensional CT stereoscopic visualization of renal masses: impact on diagnosis and patient treatment. AB - OBJECTIVE: The objective of this study was to determine whether three-dimensional reconstruction with stereoscopic display of helical CT data sets and CT angiography are useful in the examination of patients with known or suspected renal masses. CONCLUSION: Volume-rendering techniques applied to helical CT data sets coupled with three-dimensional stereoscopic imaging provide a complete examination of patients with known or suspected renal masses. Such information can help guide patient treatment and provide a single preoperative study when nephron-sparing surgery or total nephrectomy is considered. PMID- 9353451 TI - Helical CT evaluation of potential kidney donors: findings in 154 subjects. AB - OBJECTIVE: The purpose of our study was to assess renal helical CT (RHCT) as the primary imaging technique in the evaluation of potential kidney donors. SUBJECTS AND METHODS: Unenhanced and enhanced (3-mm collimation) RHCT was performed in 154 kidney donors using 125-150 ml of i.v. contrast material at an injection rate of 3 or 4 ml/sec and a pitch of 1.3-2. Scans were reconstructed at 1.5-mm intervals for a three-dimensional image. RHCT images were compared with the results of renal arteriography (RA) (50 subjects) and surgery (117 subjects). RESULTS: CT and surgical findings agreed in 95% of patients (111/117), with five cases of missed accessory arteries (all < 2 mm in diameter) and one case of a missed early division of the main artery. In the 50 subjects who underwent CT and RA, imaging revealed concordance in 96% of 100 kidneys. One small accessory artery was not detected by CT (origin from the common iliac artery). RA did not detect accessory arteries in three subjects. All 22 kidneys with early dividing main arteries (< 1.5 cm from the aortic origin) were identified by both RHCT and RA. Axial and three-dimensional CT images were complementary: five small accessory arteries were seen well only on the axial sections, whereas four early dividing arteries and two cases of renal artery stenosis were prospectively identified only on the three-dimensional images. Twenty-five renal vein anomalies were detected only by CT. In the full series of 154 subjects, nonvascular renal findings included renal calculi (n = 11), cysts (n = 12), duplicated ureters (n = 6), horseshoe kidney (n = 1), and pelvic kidney (n = 1). CONCLUSION: RHCT can be the primary imaging technique in the assessment of potential kidney donors, reducing the number of examinations as well as the risk and cost of imaging in these subjects. PMID- 9353453 TI - Role of MR imaging with transrectal coil in the evaluation of complex urethral abnormalities. AB - OBJECTIVE: The purpose of this study was to show the usefulness of MR imaging with a transrectal coil (TRC) in the management of various urethral abnormalities. This report also reveals the appearance of various postsurgical changes relating to the therapy of urethral abnormality. CONCLUSION: The high resolution images obtained with TRC MR imaging were useful in evaluating urethral abnormalities. The imaging guided therapy in most cases. In two patients, TRC MR imaging more accurately depicted abnormality than did voiding cystourethrography and transvaginal sonography. In one case, TRC MR imaging was the only imaging technique that revealed the abnormality of periurethral scarring. PMID- 9353454 TI - Imaging of renal hydatid cysts. PMID- 9353455 TI - CT findings before and after adnexal torsion: rotation of a focal solid element of a cystic adjunctive sign in diagnosis. PMID- 9353456 TI - Early detection of pneumonia in febrile neutropenic patients: use of thin-section CT. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of thin section CT for early detection of pneumonia in neutropenic patients with an unknown site of infection and normal or nonspecific findings on chest radiographs. SUBJECTS AND METHODS: Eighty-seven patients with febrile neutropenia that persisted for more than 2 days despite empiric antibiotic treatment underwent 146 prospective examinations. If findings on chest radiographs were normal (n = 126) or nonspecific (n = 20), thin-section CT (1-mm collimation, 10 mm increment) was done. If thin-section CT scans showed opacities, bronchoalveolar lavage was recommended. RESULTS: Findings on chest radiographs were nonspecific for pneumonia in 20 (14%) of 146 cases, and CT findings in those cases were suggestive of pneumonia. Microorganisms were detected in 11 of those 20 cases. Seven of the 11 cases were not optimally treated before CT diagnosis, the other four were sufficiently treated. Findings on chest radiographs and thin section CT scans were normal in 56 (38%) of 146 cases. In 70 (48%) of 146 cases, findings on chest radiographs were normal, whereas findings on thin-section CT scans were suggestive of pneumonia. Microorganisms were detected in 30 of the 70 cases. Nineteen of 30 cases were not optimally treated before CT, whereas the other 11 cases were sufficiently treated before CT. In 22 (31%) of these 70 cases, an opacity was observed on the chest radiograph during the 7 days after the CT study. Only three (5%) of 56 pneumonias occurred during the first 7 days after thin-section CT studies with normal findings (p < .005). Additional risk factors for pneumonia occurring later that were detectable on chest radiographs were poorly defined nodules (p < .05), consolidation (p < .05), and younger age (p < .05). CONCLUSION: Thin-section CT scans show findings suggestive of pneumonia about 5 days earlier than chest radiographs show suggestive findings. When thin-section CT scans show findings suggestive of pneumonia, the probability of pneumonia being detected on chest radiographs during the 7-day follow-up is 31%, whereas the probability is only 5% when the findings on the prior thin section CT scan were normal (p < .005). All neutropenic patients with fever of unknown origin and normal findings on chest radiographs should be examined with thin-section CT. PMID- 9353457 TI - Traumatic pulmonary arterial and venous pseudoaneurysms. PMID- 9353458 TI - Treatment of patients with suspected pulmonary embolism and intermediate probability lung scans: is diagnostic imaging underused? AB - OBJECTIVE: We compared patient treatment with imaging strategy in patients with clinically suspected pulmonary embolism (PE) and intermediate-probability lung scans (IPLS). MATERIALS AND METHODS: We retrospectively reviewed the medical records of 214 consecutive patients with clinically suspected PE with IPLS. RESULTS: Treatment (full anticoagulation, filter placement, or both) was given in 66 (31%) of 214 patients. Only 37% of patients were treated on the basis of definitive diagnostic imaging results. Most patients (134 [63%] of 214) were treated without an imaging diagnosis: 30 (14%) of 214 patients were treated for acute PE on clinical grounds, and the diagnosis of PE was not excluded in 104 (49%) of 214 patients. CONCLUSION: Most patients with IPLS are treated without a definitive imaging diagnosis. This lack of diagnosis may result in the overtreatment of patients who do not have acute PE or, more importantly, in the undertreatment of patients who do have acute PE. Further studies are necessary to evaluate the impact of the current management strategies on patient outcome. PMID- 9353459 TI - Hyperplasia of axillary nodes in patients undergoing immunotherapy. AB - OBJECTIVE: CT scans of the chest in 32 patients undergoing immunotherapy for metastatic renal cell carcinoma were evaluated to determine the incidence and significance of axillary lymph node enlargement during therapy. CONCLUSION: Axillary node enlargement in cancer patients undergoing immunotherapy should not be assumed to be metastatic in origin but may result from the stimulation of the immune system during treatment. PMID- 9353460 TI - Postpneumonectomy complications. PMID- 9353461 TI - Imaging of cardiac pacemakers. PMID- 9353463 TI - Findings on mammography in the axilla. PMID- 9353462 TI - Localized persistent pulmonary interstitial emphysema: CT findings with radiographic-pathologic correlation. PMID- 9353464 TI - Adaptation of an add-on stereotaxic breast biopsy unit: use of a dedicated reclinable mammography chair. PMID- 9353465 TI - An inexpensive film masking system for mammography. PMID- 9353466 TI - Simon Flexner. PMID- 9353467 TI - Breast hemangiomas in a patient with Kasabach-Merritt syndrome: imaging findings. PMID- 9353468 TI - Choroid plexus carcinoma of the lateral ventricle. PMID- 9353469 TI - Comparison of HASTE and segmented-HASTE sequences with a T2-weighted fast spin echo sequence in the screening evaluation of the brain. AB - OBJECTIVE: The purpose of this study was to evaluate the neuroradiologic application of half-Fourier acquisition single-shot turbo spin-echo (HASTE) and segmented-HASTE (s-HASTE) sequences in comparison with a T2-weighted fast spin echo sequence. MATERIALS AND METHODS: First, HASTE, s-HASTE, and fast spin-echo sequences were evaluated for blurring artifacts with a stationary phantom and for motion artifacts with a moving phantom, which repeated constant or intermittent to-and-fro motions at variable intervals. Second, 30 consecutive patients with various intracranial diseases were prospectively examined with the three sequences. Lesions were classified into four groups according to size and signal intensity on fast spin-echo MR images as follows: large hyperintense, small hyperintense, small markedly hyperintense, and hypointense lesions. Signal intensities of the lesion, putamen, and gray matter were compared with the signal intensity of white matter, and contrast-to-noise ratios were calculated. Overall image quality, conspicuity of lesions, delineation of the junction between gray matter and white matter, conspicuity of the putamen, and certain types of artifacts were evaluated qualitatively. RESULTS: In the phantom study, the HASTE sequence was least affected by motion artifacts and the fast spin-echo sequence was most affected although the images of the HASTE sequence were most degraded by blurring artifacts. In the clinical study, we found no significant differences among the three sequences for contrast-to-noise ratios or conspicuity of large hyperintense and small markedly hyperintense lesions. However, the contrast-to noise ratios of hypointense lesions and gray matter, and the conspicuity of hypointense lesions were significantly poorer for the HASTE sequence than for the fast spin-echo sequence. The contrast-to-noise ratios of small hyperintense lesions and the putamen, conspicuity of small hyperintense lesions and putamen, and delineation of the junction between gray matter and white matter were significantly poorer for HASTE and s-HASTE sequences than for the fast spin-echo sequence. Ghost artifacts, which were observed during the s-HASTE sequence, were sometimes superimposed on the image. CONCLUSION: The HASTE and s-HASTE sequences afford substantial time reduction and also decrease motion artifacts and thus have potential advantages for neuroradiologic application, especially in uncooperative or unsedated children. The s-HASTE sequence may be preferable to the HASTE sequence because of fewer blurring artifacts and higher T2 contrast. However, small hyperintense and hypointense lesions may be overlooked when HASTE and s-HASTE sequences are used. PMID- 9353470 TI - CT angiography: source images and postprocessing techniques in the detection of cerebral aneurysms. PMID- 9353471 TI - Black blood MR angiography using multislab three-dimensional TI-weighted turbo spin-echo technique: imaging of intracranial circulation. PMID- 9353472 TI - Imaging appearance of pachymeningeal tuberculosis. AB - OBJECTIVE: The purpose of our study was to examine imaging findings in patients with pachymeningeal tuberculosis. Imaging studies of seven patients with pachymeningeal tuberculosis were retrospectively reviewed. The diagnosis had been established on the basis of histopathology in three patients and response to antitubercular treatment in four patients. CONCLUSION: Tuberculosis can lead to localized or diffuse involvement of the pachymeninges. Most of the focal lesions were seen as en plaque, homogeneous, uniformly enhancing, dural-based masses. The lesions appeared hyperdense on plain CT scans, isointense to brain parenchyma on T1-weighted MR images, and isointense to hypointense on T2-weighted MR images. One patient had diffuse sheet-like thickening of the pachymeninges in the right hemicranium, involving both the supratentorial and infratentorial compartments. PMID- 9353473 TI - Enhanced MR imaging of hypertrophic pachymeningitis. AB - OBJECTIVE: This report illustrates the contrast enhancement characteristics on MR imaging of three patients with hypertrophic pachymeningitis and provides an explanation for the observed imaging findings. CONCLUSION: A differential pattern of enhancement, consisting of intense enhancement of the peripheral margin of the abnormal pachymeninges, was present in all cases. In two patients, much of the remaining abnormal pachymeninges did not enhance at all. On the basis of the microscopic pathology of hypertrophic pachymeningitis, the physiology of normal meningeal enhancement on MR imaging, and the described MR appearance of other pachymeningeal lesions, this differential pattern of enhancement should strongly suggest the diagnosis of hypertrophic pachymeningitis. PMID- 9353474 TI - Cerebellopontine angle lipomas, multiple pigmented nevi, and temporal lobe hypoplasia: a new neurocutaneous syndrome? PMID- 9353475 TI - Radiologic placement of subcutaneous infusion chest ports for long-term central venous access. AB - OBJECTIVE: The technical success and complications associated with radiologic placement of subcutaneous implantable chest ports for long-term central venous access were evaluated. MATERIALS AND METHODS: Between May 1, 1996, and December 31, 1996, 80 chest ports were placed in 80 consecutive patients using the right internal jugular vein as the preferred access route. All procedures occurred in interventional radiology suites with patients receiving conscious sedation. Both sonography and fluoroscopy were used for venipuncture and to guide port insertion. Follow-up was obtained by the clinical service and by performing chart reviews electronically. RESULTS: Technical success was 100%, and follow-up was obtained in all patients. One procedural complication occurred that was unrelated to actual catheter placement. Mean catheter use was 155 days (total, 12,168 days; range, 18-303 days). Confirmed catheter-related infection rate was 3%, or 0.016 per 100 access days; symptomatic catheter-related central venous thrombosis rate was 1%, or 0.008 per 100 access days; and 5% of catheters were removed prematurely. No instances of hematoma formation, catheter tip migration or malposition, symptomatic air embolism, spontaneous catheter fracture, or pneumothorax were found. CONCLUSION: With the benefit of both sonographic and fluoroscopic guidance, subcutaneous implantable chest ports can be inserted by radiologists with equal or lower complication rates than those reported in surgical series. Image-guided insertion of chest ports should replace rather than supplement unguided placement. PMID- 9353477 TI - In-phase and out-of-phase MR imaging of bone marrow: prediction of neoplasia based on the detection of coexistent fat and water. AB - OBJECTIVE: The purpose of this study was to determine if gradient-echo MR imaging with TEs selected with fat and water in phase and out of phase can help predict the likelihood of neoplastic or nonneoplastic lesions in bone marrow. SUBJECTS AND METHODS: Thirty consecutive patients with 31 suspected bone marrow lesions underwent MR imaging, including two spoiled gradient-echo sequences identical in all parameters except TE, which was chosen such that fat and water were either in phase or out of phase. Relative ratios of the abnormal bone marrow signal intensity and a control site on the in-phase and out-of-phase images were expressed. The images were also assessed independently by two reviewers who were unaware of the patients' identities and clinical histories. Reviewers assessed decreased marrow signal intensity relative to control sites on the out-of-phase and in-phase images. Pathologic confirmation was obtained in 16 patients (17 lesions); the remainder of patients had either established diagnoses or determination of benignity based on stability of findings at 1 year. Relative ratios were compared with the Student's t test and receiver operating characteristic (ROC) curve analysis, and the reviewers' scores were evaluated with ROC curve analysis. RESULTS: The relative signal-intensity ratios were 1.03 +/- 0.13 for the neoplastic group and 0.62 +/- 0.13 for the nonneoplastic group (p < .0001). ROC curve analysis of the signal-intensity ratios showed a z-score of .99. A ratio cutoff value of 0.81 resulted in a 95% sensitivity and a 95% specificity for detection of neoplasm. Both reviewers achieved 100% sensitivity and 94-100% specificity for detection of neoplasms. CONCLUSION: In-phase and out of-phase gradient-echo MR imaging of bone marrow signal-intensity abnormalities can help predict the likelihood of neoplastic or nonneoplastic lesions. PMID- 9353476 TI - Wallstent deployment to salvage dialysis graft thrombolysis complicated by venous rupture: early and intermediate results. AB - OBJECTIVE: The feasibility of deploying Wallstents to treat venous rupture occurring during dialysis graft thrombolysis was determined. SUBJECTS AND METHODS: Between June 24, 1994, and February 19, 1997, 23 patients with venous rupture attributed to balloon angioplasty during dialysis graft thrombolysis were treated by Wallstent deployment across the area of rupture. Twenty-one ruptures occurred in peripheral veins and two occurred in central veins. Follow-up was provided by the clinical service at our institution and by electronic review of patients' charts. RESULTS: Stent placement allowed completion of graft thrombolysis in all 23 patients. Complications were limited to four moderate sized hematomas of the arm, and a single pseudoaneurysm developed 6 months after stent placement in one 27-year-old patient. The primary patency rate of stents was 52% at 60 days, 26% at 180 days, and 11% at 360 days. The secondary patency rate was 74% at 60 days, 65% at 180 days, and 56% at 360 days. CONCLUSION: Treatment of venous ruptures using Wallstents is a safe alternative to intentional graft thrombosis. The patency rates of these devices are similar to those of venous stents placed for other indications. PMID- 9353478 TI - Superficial soft-tissue masses suggestive of recurrent malignancy: sonographic localization and biopsy. AB - OBJECTIVE: We investigated the usefulness of high-resolution sonography to localize superficial soft-tissue masses and to guide needle sampling for recurrent malignancy. MATERIALS AND METHODS: High-resolution sonography (10-MHz) was used to locate and guide needle sampling of 16 palpable and eight impalpable superficial masses suggestive of recurrent malignancy in 23 patient (12 men, 11 women; 34-85 years old). After detection, 22 (92%) of the masses were immediately sampled by fine-needle aspiration with 18- to 25-gauge needles and two (8%) were sampled by a 20-gauge core gun. RESULTS: Diagnostic material was obtained without complication from all 24 masses and proved positive for recurrent disease in 13 (54%). Ten (63%) of 16 palpable and three (38%) of eight impalpable masses proved positive for recurrent malignancies. One third of superficial soft-tissue masses were detected by imaging only, and of the masses not revealed on imaging, three (23%) of 13 were the site of first recurrence. Most nonnodal superficial masses (8/13) were benign, unlike the lymph nodes, of which three (27%) of 11 were benign. CONCLUSION: Not all early recurrent malignancies within the skin and subcutaneous tissues are detected by clinical examination. High-resolution sonography provided us with a rapid, safe, and accurate means of localizing and then guiding needle biopsies of superficial soft-tissue masses suggestive of recurrent malignancy. PMID- 9353479 TI - MR arthrography of the shoulder: rethinking traditional imaging procedures to meet the technical requirements of MR imaging guidance. AB - OBJECTIVE: The purpose of this study was to determine the feasibility of and the appropriate technique for performance of MR imaging-guided arthrography of the shoulder. SUBJECTS AND METHODS: Thirty-eight MR imaging-guided glenohumeral joint punctures were performed using an open C-arm scanner with a vertically oriented magnetic field, adapted for interventional procedures. Two different approaches to the shoulder were used: a modification of the traditional anterior approach (seven procedures), and an anterosuperior approach (31 procedures) mimicking the anterior arthroscopy portal. The average procedure duration was determined. A retrospective review of needle mediolateral and anterioposterior position was determined for the anterosuperior approaches. RESULTS: Average procedure duration was 21 min for the anterior approach and 12 min for the anterosuperior approach. Subjectively, needle conspicuity was minimal with the anterior approach, contributing to prolonged imaging times. Needle visualization was much improved with the anterosuperior approach. Nine of the 31 anterosuperior procedures involved inadvertent injection of the subacromial or subdeltoid bursa. At the time of retrospective review, the needle was too laterally or too anteriorly positioned in six of these nine patients. CONCLUSION: With consideration of the technical demands of MR imaging guidance for interventional procedures, MR imaging-guided arthrography of the shoulder is feasible. The traditional radiologic approach to the shoulder must be modified to provide adequate visualization of the needle. The anterosuperior approach meets this needs. PMID- 9353480 TI - Re: MR imaging of vertebral osteomyelitis revisited. PMID- 9353482 TI - Emphysema and chronic obstructive pulmonary disease. PMID- 9353481 TI - Re: Placement of long-term central venous catheters in outpatients. PMID- 9353483 TI - "Bulitas": penile implants to enhance sexual prowess. PMID- 9353484 TI - Multiple epidermoid splenic cysts: unusual findings. PMID- 9353485 TI - Small-bowel lymphoma complicating long-standing Crohn's disease. PMID- 9353486 TI - Choroid plexus papilloma: detection using color Doppler sonography. PMID- 9353487 TI - Impact of core biopsy on the surgical management of impalpable breast cancer: another look at margins. PMID- 9353488 TI - Perforation of the colon during barium enema examination. PMID- 9353489 TI - Mediastinal emphysema with Pneumocystis carinii pneumonia in AIDS. PMID- 9353490 TI - Metastatic renal cell carcinoma to the gallbladder: color Doppler sonography and CT findings. PMID- 9353491 TI - The relation between fetal malnutrition and chronic disease in later life. PMID- 9353492 TI - The use of statins: a case of misleading priorities? PMID- 9353493 TI - Genetic diagnosis before implantation. PMID- 9353494 TI - Hunger strikes. PMID- 9353495 TI - New study reports that diet is critical to cancer prevention. PMID- 9353496 TI - Doctors volunteer to be guinea pigs for AIDS vaccine. PMID- 9353497 TI - FDA insists on more women in drug trials. PMID- 9353498 TI - When life saving treatment should be withdrawn in children. PMID- 9353499 TI - Government reviews law on "posthumous conceptions". PMID- 9353500 TI - Health care in China is highly inequitable. PMID- 9353501 TI - Dutch and Swiss support heroin on prescription. PMID- 9353502 TI - Mother's weight in pregnancy and coronary heart disease in a cohort of Finnish men: follow up study. AB - OBJECTIVE: To determine whether restricted growth in utero is associated with an increased risk of coronary heart disease are among men in Finland, where rates of the disease are among the highest in the world. DESIGN: Follow up study. SETTING: Helsinki, Finland. SUBJECTS: 3302 men born in Helsinki University Central Hospital during 1924-33 who went to school in the city of Helsinki and were resident in Finalnd in 1971. MAIN OUTCOME MEASURES: Standardised mortality ratios for coronary heart disease. RESULTS: Men who were thin at birth, with low placental weight, had high death rates from coronary heart disease. Men whose mothers had a high body mass index in pregnancy also had high death rates. In a multivariate analysis the hazard ratio for coronary heart disease was 1.37 (95% confidence interval 1.20 to 1.57) (P < 0.0001) for every standard deviation decrease in ponderal index at birth and 1.24 (1.10 to 1.39) (P = 0.0004) for every standard deviation increase in mother's body mass index. The effect of mother's body mass index was restricted to mothers of below average stature. CONCLUSION: These findings suggest a new explanation for the epidemics of coronary heart disease that accompany Westernisation. Chronically malnourished women are short and light and their babies tend to be thin. The immediate effect of improved nutrition is that women become fat, which seems to increase the risk of coronary heart disease in the next generation. With continued improvements in nutrition, women become taller and heavier; their babies are adequately nourished; and maternal fatness no longer increases the risk of coronary heart disease, which therefore declines. PMID- 9353503 TI - A meta-analysis of cigarette smoking, bone mineral density and risk of hip fracture: recognition of a major effect. AB - OBJECTIVE: To determine the magnitude and importance of the relation between smoking, bone mineral density, and risk of hip fracture according to age. DESIGN: Meta-analysis of 29 published cross sectional studies reporting the difference in bone density in 2156 smokers and 9705 non-smokers according to age, and of 19 cohort and case-control studies recording 3889 hip fractures reporting risk in smokers relative to non-smokers. RESULTS: In premenopausal women bone density was similar in smokers and non-smokers. Postmenopausal bone loss was greater in current smokers than non-smokers, bone density diminishing by about an additional 2% for every 10 year increase in age, with a difference of 6% at age 80. In current smokers relative to non-smokers the risk of hip fracture was similar at age 50 but greater thereafter by an estimated 17% at age 60, 41% at 70, 71% at 80, and 108% at 90. These estimates of relative risk by age, derived directly from a regression analysis of the studies of smoking and hip fracture, were close to estimates using the difference in bone density between smokers and non-smokers and the association between bone density and risk of hip fracture. The estimated cumulative risk of hip fracture in women in England was 19% in smokers and 12% in non-smokers to age 85; 37% and 22% to age 90. Among all women, one hip fracture in eight is attributable to smoking. Limited data in men suggest a similar proportionate effect of smoking as in women. The association was not explained by smokers being thinner, younger at menopause, and exercising less nor by actions of smoking on oestrogen, but smoking may have a direct action on bone. CONCLUSIONS: Hip fracture in old age is a major adverse effect of smoking after the menopause. The cumulative excess bone loss over decades is substantial, increasing the lifetime risk of hip fracture by about half. PMID- 9353504 TI - Beer binging and mortality: results from the Kuopio ischaemic heart disease risk factor study, a prospective population based study. AB - OBJECTIVE: To examine the association between beer binging (regular sessions of heavy beer drinking) and mortality. DESIGN: Prospective population based study with the baseline assessment of level of alcohol intake (dose), by type of drink and drinking pattern, previous and existing diseases, socioeconomic background, occupational status, involvement in organisations during leisure time, physical activity in leisure time, body mass index, blood pressure, serum lipids and plasma fibrinogen concentration, during an average of 7.7 years' follow up of mortality. SETTING: Finland. SUBJECTS: A population sample of 1641 men who consumed beer who were aged 42, 48, 54, or 60 years at baseline. MAIN OUTCOME MEASURES: All cause mortality, cardiovascular mortality, death due to external causes, fatal myocardial infarctions. RESULTS: The risk of death was substantially increased in men whose usual dose of beer was 6 or more bottles per session compared with men who usually consumed less than 3 bottles, after adjustment for age and total alcohol consumption (relative risk 3.01 (95% confidence interval 1.54 to 5.90) for all deaths; 7.10 (2.01 to 25.12) for external deaths; and 6.50 (2.05 to 20.61) for fatal myocardial infarction). The association changed only slightly when smoking, occupational status, previous diseases, systolic blood pressure, low density lipoprotein and high density lipoprotein cholesterol concentration, plasma fibrinogen concentration, body mass index, marital status, leisure time physical activity, and involvement in organisations were controlled for. CONCLUSION: The pattern of beer binging is associated with increased risk of death, independently of the total average consumption of alcoholic drinks. The relation is not explained by known behavioural, psychosocial, or biological risk factors. Death due to injuries and other external causes is overrepresented among beer bingers, but a strong association with fatal myocardial infarction suggests that the pathway may also involve other acute triggers of severe health events. PMID- 9353505 TI - Time since childbirth and prognosis in primary breast cancer: population based study. AB - OBJECTIVE: To investigate whether time since birth of last child was of prognostic importance in women with primary breast cancer. DESIGN: Retrospective cohort study based on a population based database of breast cancer diagnoses with detailed information on tumour characteristics, treatment regimens, reproductive factors, and vital status. SETTING: Denmark. SUBJECTS: 5652 women with primary breast cancer aged 45 years or less at the time of diagnosis. MAIN OUTCOME MEASURES: 5 and 10 year survival; relative risk of dying. RESULTS: Women diagnosed in the first 2 years after last childbirth had a crude 5 year survival of 58.7% and 10 year survival of 46.1% compared with 78.4% and 66.0% for women whose last childbirth was more than 2 years before their diagnosis. After adjustment for age, reproductive factors, and stage of disease (tumour size, axillary nodal status, and histological grading), a diagnosis sooner than 2 years since last childbirth was significantly associated with a poor survival (relative risk 1.58, 95% confidence interval 1.24 to 2.02) compared with women who gave birth more than 5 years previously. Further analyses showed that the effect was not modified by age at diagnosis, tumour size, and nodal status. CONCLUSIONS: A diagnosis of breast cancer less than 2 years after having given birth is associated with a particularly poor survival irrespective of the stage of disease at debut. Therefore, a recent pregnancy should be regarded as a negative prognostic factor and should be considered in counselling these patients and in the decisions regarding adjuvant treatment. PMID- 9353506 TI - Audit of child protection procedures in accident and emergency department to identify children at risk of abuse. PMID- 9353507 TI - Cerebral and cerebellar atrophy on serial magnetic resonance imaging in an initially symptom free subject at risk of familial prion disease. PMID- 9353508 TI - Effect of inhaled corticosteroids on episodes of wheezing associated with viral infection in school age children: randomised double blind placebo controlled trial. AB - OBJECTIVES: To determine the effect of regular prophylactic inhaled corticosteroids on wheezing episodes associated with viral infection in school age children. DESIGN: Randomised, double blind, placebo controlled trial. SETTING: Community based study in Southampton. SUBJECTS: 104 children aged 7 to 9 years who had had wheezing in association with symptoms of upper and lower respiratory tract infection in the preceding 12 months. INTERVENTIONS: After a run in period of 2-6 weeks children were randomly allocated twice daily inhaled beclomethasone dipropionate 200 micrograms or placebo through a Diskhaler for 6 months with a wash out period of 2 months. Children were assessed monthly. MAIN OUTCOME MEASURES: Forced expiratory volume in 1 second (FEV1); bronchial responsiveness to methacholine (PD20); percentage of days with symptoms of upper and lower respiratory tract infection with frequency, severity, and duration of episodes of upper and lower respiratory symptoms and of reduced peak expiratory flow rate. RESULTS: During the treatment period there was a significant increase in mean FEV1 (1.63 v 1.53 1; adjusted difference 0.09 1 (95% confidence interval 0.04 to 0.14); P = 0.001) and methacholine PD20 12.8 v 7.2 mumol/l; adjusted ratio of means 1.7 (1.2 to 2.4); P = 0.007) in children receiving beclomethasone dipropionate compared with placebo. There were, however, no significant differences in the percentage of days with symptoms or in the frequency, severity, or duration of episodes of upper or lower respiratory symptoms or of reduced peak expiratory flow rate during the treatment period between the two groups. CONCLUSIONS: Although lung function is improved with regular beclomethasone dipropionate 400 micrograms/day, this treatment offers no clinically significant benefit in school age children with wheezing episodes associated with viral infection. PMID- 9353510 TI - ABC of palliative care. Difficult pain problems. PMID- 9353509 TI - Recent advances in endocrine therapy of breast cancer. PMID- 9353511 TI - Climate change and health: implications for research, monitoring, and policy. PMID- 9353512 TI - Resource allocation to health authorities: the quest for an equitable formula in Britain and Sweden. PMID- 9353513 TI - Antibiotics as initial treatment for children with acute otitis media. Use of odds ratio in calculation of number needed to treat was inappropriate. PMID- 9353514 TI - Antibiotics as initial treatment for children with acute otitis media. Diagnostic criteria need to be defined. PMID- 9353515 TI - Antibiotics as initial treatment for children with acute otitis media. Abandoning treatments that you have used for years in difficult. PMID- 9353516 TI - Relative risks are inflated in published literature. PMID- 9353517 TI - Study confirms results of systematic review of care in stroke unit. PMID- 9353518 TI - Modern treatment for internal haemorrhoids. Day surgery offers permanent cure. PMID- 9353519 TI - Modern treatment for internal haemorrhoids. Rubber band ligation is effective and efficient. PMID- 9353520 TI - Author preferred to cite substantive references rather than meeting abstracts. PMID- 9353521 TI - Effect of deprivation on general practitioners' referral rates. Study should have used deprivation index that is independent of age. PMID- 9353522 TI - Effect of deprivation on general practitioners referral rates. Jarman score measures workload not deprivation. PMID- 9353523 TI - Effect of deprivation on general practitioners referral rates. Analyses should take age and sex into account. PMID- 9353524 TI - Mental health emergencies. Details of studies of zuclopenthixol acetate are needed. PMID- 9353525 TI - Mental health emergencies. Patients need to be monitored when given rapid tranquillisation. PMID- 9353526 TI - Mental health emergencies. Zuclopenthixol acetate is given by intramuscular injection, not intravenously. PMID- 9353527 TI - Mental health emergencies. Caution is needed with rapid tranquillisation protocol. PMID- 9353528 TI - Data on results of using different antiepileptic drugs do exist. PMID- 9353529 TI - Accident and emergency departments should not be considered places of safety. PMID- 9353530 TI - Neonatal mortality of low birthweight babies is increased in mothers with higher education. PMID- 9353531 TI - Acupuncturists have begun initiative to agree standards to improve safety. PMID- 9353532 TI - Comparison of different selective methods for detection of Listeria species in surface water. AB - Tests were carried out to evaluate the efficiency of various combinations of selective enrichment and plating techniques using pure cultures of Listeria monocytogenes, Listeria seeligeri and Listeria innocua in suspension and samples of surface water. The best yields for the various Listeria spp. were obtained after a single passage in LEB (Oxoid) or LEB Buffered (Oxoid) and after using Palcam and Oxford agar (Oxoid). Palcam agar was, however, shown to be the most efficient means of detecting the Listeria spp. in the natural water samples. PMID- 9353533 TI - Comparison of BGB-MUG and LSTB-MUG in microbiological surveillance of recreational waters. AB - Recreational water surveillance is an important tool to prevent health hazards for the population. Therefore distinct guide and imperative values for fecal indicators are listed in the EC directive about water quality control. The detection methods, however, give laboratories some room to choose their own method, which has led to difficulties in the comparability of results. In 1989 an ad-hoc working group of the coastal countries of Germany established detection methods, which by now are obligatory for these countries. Fecal and total coliforms (FC and TC) are detected by a triplicate mpn-procedure using brilliant green-bile-lactose broth supplemented with tryptophane and 4-methylumbelliferyl beta-D-glucuronide (BGB-MUG) as selective medium. Gas-, fluorescence- and indole positive cultures are considered fecal coliform-positive. In the last years rises in TC but not in FC counts were observed in fresh waters. A study was carried out to evaluate the official method in another bathing season, to determine bacterial species leading to false-positive TC cultures and to compare BGB-MUG with laurylsulphate-tryptophane-MUG (LSTB-MUG). Water samples of different salinities and nutrient input were collected in weekly intervals from April to October. FC and TC concentrations were determined and all TC-positive cultures were differentiated further. The FC counts obtained by enrichment in BGB-MUG or LSTB MUG were nearly identical, the rate of fluorescence-positive, indole-negative tubes being approximately 0.6%. Differentiation of FC-negative cultures showed a false-negative rate of 2.87% for BGB-MUG and of 8% for LSTB-MUG. During the summer months TC counts in BGB-MUG exceeded FC counts by far at most of the sampling sites. This effect was much less pronounced in LSTB-MUG; the difference between both enrichment media being significant. Differentiation of presumptive TC from BGB-MUG resulted in a high percentage of Aeromonas spp. in fresh waters. LSTB-MUG was clearly more selective for TC than BGB-MUG, but still with an average of 10% of the test tubes being false TC-positive (BGB-MUG 46%). The sensitivity of BGB-MUG was below 60% (LSTB-MUG 89%). LSTB-MUG should be preferred as enrichment medium in mpn-examination of recreational water, if no further differentiation is carried out. The selectivity for TC is better than in BGB-MUG and the only slight inhibitory effects can be tolerated. PMID- 9353534 TI - Evaluation of MUG-supplemented media for the detection of E. coli in recreational water surveillance. AB - Four media containing 4-methylumbelliferyl-beta-D-glucuronide were evaluated as a non-confirmatory procedure for E. coli detection in recreational water surveillance. The media included ECD-Agar for membrane filtration and laurylsulphate-tryptose, brilliant-green-bile and lactose as broth media in a three tube most probable number procedure. From six representative water sites, samples were collected weekly over a typical summer season (17.05-27.09.1994) and processed as parallels, using each media at two different incubation temperatures (36 degrees/44 degrees C). Results showed that incubation temperature had no impact on E. coli counts. Each media at a given temperature could be regarded as individual enrichment procedure. None of these enrichment procedures showed a constant and predictable higher sensitivity during the sampling period at all sites compared to the others tested. For parallel results, the rate of agreement, based upon EC-guideline (76/160/EWG) staging of recreational water quality, was 85% for membrane filtration and 75% for the MPN-procedure results. Marked differences could be observed in false-positive specificity showing correlation to the selective characteristics of the media. Subsequently lactose-broth at 44 degrees C performed worst with 30% non verifiable results, while ECD-agar and laurysulphate-tryptose-broth, both at 44 degrees C, had a nearly 100% confirmation rate. Thus, combining high specificity with no lack in sensitivity these two MUG-supplemented media seem to be best suited for E. coli detection in routine recreational water surveillance. PMID- 9353535 TI - Bacteriocin typing of Klebsiella spp. isolated from different sources. AB - To evaluate whether clinical Klebsiella isolates differ from nonclinical strains with respect to bacteriocin susceptibility patterns, a total of 452 Klebsiella pneumoniae and K. oxytoca strains isolated from different sources were examined. Bacteriocin typing was done by a modification of the scrape-and-point method, using a set of eight producer strains. 96% of the strains were typable. Forty-one different bacteriocin susceptibility patterns were observed. While two thirds of the K. oxytoca isolates belonged to only three different bacteriocin types, the K. pneumoniae strains showed a more heterogeneous distribution of patterns. No differences in pattern distribution were observed between isolates from clinical, fecal, or environmental sources. Certain bacteriocins showed a very broad spectrum of activity; e.g. 93% of all isolates were susceptible to bacteriocin type 3. The results suggest that nonclinical Klebsiella strains do not show other bacteriocin susceptibility types than clinical isolates do. PMID- 9353536 TI - Comparison study on three protocols used to concentrate poliovirus type 1 from drinking water. AB - The efficiency of three techniques used to concentrate enteric viruses from water media and based on adsorption-elution on glass are tested. The techniques are adsorption on glass wool (GW) at the natural pH of the water and adsorption on glass powder using acidified water (pH 3.5). In the second case, two devices are used the classical apparatus (CGP) and the modified apparatus (MGP). A solution of glycine 0.05 M--3% beef extract pH 9.5 is used in all three techniques to perform the elution. The sensitivity of the above concentration methods is assayed with samples of 20 liters of tap water artificially contaminated with a known quantity of poliovirus type 1 (10(1) to 10(7) MPNCU [20 L]-1). The resulting concentrates are inoculated to BGM cell cultures and tittered according to the MPN technique. The study demonstrated that the recovery rate increased with the viral concentration of the samples with maximum efficiency reaching 81% for GW, 89% for CGP and 99% for MGP. A Wilcoxon test performed on paired samples and on the overall results with all three methods. Significant differences were demonstrated leading to the ranking of the techniques in the growing order of sensitivity GW, CGP and MGP. These finding were confirmed using a fitting technique according to the algorithm of Marquardt. PMID- 9353537 TI - Asthma and exacerbation of chronic bronchitis: sentinel and environmental data in a time series analysis. AB - To see whether effects of air pollutants and other environmental factors on the respiratory tract can be detected by the Swiss sentinel reporting system, two years' data of asthma bronchiale and exacerbation of chronic bronchitis were analyzed. On average, 16 cases of asthma and 9 cases of bronchitis were reported per week. The respective figures expressed as mean percentages of all consultations were 0.12% and 0.065%. Data of SO2, NO2, ozone and total suspended particles were used to measure air pollution. Additionally, meteorologic parameters such as air temperature and atmospheric pressure were used, as well as the appearance of the most important pollen groups in Switzerland: grass, birch tree and mugwort. Environmental data were summarized using the mean or sum of all measuring stations. Autocorrelations in the time series were accounted for statistically. Our analysis could not establish any relationship between reports of asthma or exacerbation of chronic bronchitis and air pollutants or other environmental data. This result, which is partly contradicted by the literature, could be explained by low numbers of reports due to patient's self administration of medication and an imprecise determination of true exposure. PMID- 9353538 TI - Internal and external tetrachloroethene exposure of persons living in differently polluted areas of Northrhine-Westphalia (Germany). AB - An epidemiological study was performed to measure the internal and external tetrachloroethene exposure of persons living in two differently polluted areas of Northrhine-Westphalia (Germany). Tetrachloroethene concentrations were determined in venous blood samples of 5- to 7-year-old children (n = 81) and 55-year-old women (n = 91) living in Essen, an industrial city located in the Ruhr area. 103 children und 131 women of the same age living in Borken, a small town north of the Ruhr area, served as reference group. Outdoor air samples were collected on passive samplers (sampling period: 4 weeks) from 70 measurement points per study area (about 2 km2, mean distance 100 m). In the course of a year these measurements were repeated three times to cover seasonal variations. Parallel to the outdoor measurement periods, indoor air concentrations were determined in the homes of those women from Essen and Borken, who donored a blood sample. Tetrachloroethene levels in blood were generally low with a geometric mean of 0.05 microgram/L in women and 0.021 microgram/L in children. Nevertheless, children and women living in the industrial area were found to have significantly higher tetrachloroethene levels in blood than those of the reference group. In both study areas blood levels of women exceeded those of children by a factor of 2. Participants living in the neighbourhood of a dry-cleaning shop had distinctly elevated blood levels. The same applied to persons who stored dry-cleaned clothes at home. Like the internal exposure, external exposure was also higher in Essen than in Borken. In both areas tetrachloroethene concentrations indoors exceeded those outdoors. Outdoor tetrachloroethence concentrations were significantly increased during the cold season, while the opposite was true for indoor levels. The correlation between indoor and outdoor exposure was found to be significant, while those between blood levels and outdoor exposure became only significant when people living next to a dry-cleaning shop were excluded. No significant relationship was observed between blood and indoor tetrachlorethene levels. It is concluded that the higher tetrachloroethene blood levels of the urban population result from the higher atmospheric concentrations in industrial areas with tetrachloroethene emitting sources like metal and textile industry. The fact that indoor air tetrachloroethene levels exceeded those outdoors can only be explained by the presence of additional indoor sources. Provided that women spend on average more time indoors than children the higher indoor air concentrations may be the reason for the higher blood tetrachloroethene levels found in women. Persons living near a dry-cleaning shop or storing dry-cleaned clothes at home showed a higher internal and external exposure to tetrachloroethene than other persons. In individual cases it can by far exceed the average exposure of the general population, so that health impairments can not be generally excluded. PMID- 9353539 TI - Hair analysis in environmental medicine. AB - Hair analysis comprises the determination of minerals, trace elements and drugs. It is applied in Germany with increasing frequency in the recently established field of environmental medicine for biological monitoring of the internal metal/metalloid exposure. Besides a number of advantages hair analysis is impaired by the difficulty-to distinguish between endogenous and exogenous sources of metals in hair. Except for methylmercury, there are no critical limit values for trace elements in hair available. However, valid reference values for some metals and for nicotine in hair have been recently presented in the German Environmental Survey. The significance of selected substances in hair are as follows: Aluminium in hair is of no value in environmental medicine. For assessment of cadmium and inorganic arsenic exposure hair analysis is only suitable as a screening method based on large populations. Monitoring of lead in hair is a valuable screening method also for small groups, especially for children. Based on toxicokinetics and under consideration of practicability the optimal biomarker of methylmercury exposure is the hair concentration. For other mercury compounds hair analysis is of lower significance. Nicotine and cotinine measurements in hair provide a practical and proper method for estimating environmental tobacco smoke exposure and to validate smoker status in epidemiological studies. PMID- 9353540 TI - Evaluation of a new version of the EnviroAmp Legionella kit for the detection of legionellae in water samples by the polymerase chain reaction. AB - The detection of Legionella sp. in various types of environmental water samples was evaluated using a revised version of the EnviroAmp Legionella kit (Perkin Elmer), which is based on the polymerase chain reaction (PCR). The new kit employs a modified protocol for the pretreatment of water samples, which is designed to further reduce the inhibitory action of some samples to the PCR. The results of the new kit were compared with those of culture. Seventy-four water samples were examined. Of these samples, 51 were taken from hospital water systems, 16 were household water samples, 5 were from surface water, and 2 were rain water. These samples were examined both by PCR and by culture on MWY Legionella selective agar. Of the 74 samples, 46 (62%) were positive by both test systems. Twenty-five samples (34%) were positive by PCR, but negative by culture. While 3 samples (4%) were negative by both test methods, no sample was negative by PCR and positive by culture. Statistical analysis revealed high positive predictive values of PCR, when L. pneumophila was detected or when the genus Legionella was detected at high intensity. Negative predictive values were high, when the PCR was negative or only weakly positive for the genus Legionella. PMID- 9353541 TI - Effect of water softening and heating on microbial contamination of dental unit systems. AB - This study regards the quality of the water used in 4 types of dental units making use of softened and heated water, softened but non-heated water, non softened but heated water and non-softened and non-heated water. The samples were taken from the incoming tap water, from oral rinsing cup, the air-water syringe and the ultrasound descaling hand-piece. The results showed how the water underwent a notable growth in bacteria during its passage within the circuits of the units, reaching heterotrophic total counts greatly exceeding the guidelines set down by Italian laws regarding drinking water. While the influence of softening was evident, the bacteria in the samples taken from descaling handpiece, where there is more stagnation, found excellent growing conditions also at high temperatures. In the softened and heated waters a notable growth of Pseudomonas aeruginosa was found and it is likely that this was encouraged by the combined effect of the softening and heating. As far as the origin of the contamination is concerned, the bacteria present in the water systems seem to have come from the incoming water. PMID- 9353542 TI - Evaluation of disinfection treatment systems for municipal wastewater reclamation and reuse. AB - The efficiency of a number of tertiary treatment systems--filtration, ozonation, chlorination with low levels (TRC < 0.2 ppm) and high levels (TRC < 1 ppm) of residual chlorine--in the disinfection of secondary effluent was assessed in a purification plant treating mixed sewage of municipal (83%) and industrial mainly textile origin (17%). Maximum purification effect was observed when, following secondary treatment with biological oxidation, the sewage was submitted to combined filtration--ozonation treatment (reduction in bacterial indicators of from 4.9 to 7.2 log10 units) or with chlorination with high levels of residual chlorine (reduction in the bacterial indicators of between 2.8 and 4.6 log10 units). However, only ozonation reduced viral indicators with respect to inflow sewage by more than 3 log10 units, the limit considered acceptable for a biological treatment system with supplementary tertiary disinfection treatment. Ozonation however did not complete control all the biological forms present in the sewage, in particular the viruses, present in 36% of ozonized samples at concentrations of from 1 to 480 PFU/100 mL. Ozonation and high-concentration chlorination do not seem to be unfavorably influenced by wastes from laundry and deyng processing; achieving a complete decolorization of the treated effluent, they prove to the suitable treatments for mixed sewage of municipal and industrial mainly textile origin. PMID- 9353543 TI - Comparative disinfection of secondary-treated sewage with chlorine dioxide and bromine chloride. AB - A comparison was made of the inactivation rates of Arcobacter butzleri, coliphages, total coliforms, fecal coliforms, fecal streptococci and heterotrophic plate count in secondary sewage effluent using chlorine dioxide (2 and 4 ppm) and bromine chloride (4 or 8 and 12 ppm) as disinfecting agents. Using these doses the ClO2 gave higher reduction percentages (on average more than 99% at 4 ppm) than those obtained with BrCl. The average values of the fecal indicators are well within the legal limits. Arcobacter butzleri was more sensitive to the disinfectants than other bacteria while fecal streptococci were seen to be more resistant. From the chemical point of view no differences were seen between the two disinfectants except that the action of ClO2 was stronger regarding BOD5 than that of BrCl. With the exception of dichloromethane, the concentration of volatile halogenated compounds showed little variation and values were often lower than detection limits. PMID- 9353544 TI - Occurrence and detection of viable Listeria in food scrap compost. AB - Listeria species (L. innocua, L. ivanovii, L. seeligeri, and L. grayi) were readily detected in food scraps by Nucleic Acid Hybridization (NAH) probes using a standard Listeria selective medium (UVM-1) at ambient temperature. Various food scrap compost recipes artificially contaminated with Listeria at 10(7) cells per gram wet weight were composted in thermally insulated bench scale reactor vessels. These Listeria were not detected when the compost temperature became elevated. Different isolation methods for the Listeria showed this result to be a false negative occurring apparently because the heat stressed Listeria were unable to survive in the selective medium (UVM-1). Once incubated at 37 degrees C in Universal Listeria medium (ULM), the Listeria were detectable for a short period in compost at temperatures as high as 64 degrees C. PMID- 9353545 TI - Prevalence of Listeria monocytogenes and other listerias in Italian-made soft cheeses. AB - The frequency of L. monocytogenes and other listerias was determined in different types of Italian-made soft cheeses purchased from retail outlets (shops and supermarkets) located in different areas and different towns of central Italy. Of the 164 examined samples, eight proved to be positive for L. monocytogenes (4.9%), seven strains belonged to serotype 1, and one strain to serotype 4. Thirty-six samples were positive for the presence of other listeria species (22%); of these, L. innocua was prevalent (72% of positive samples). The cheeses bought in supermarkets displayed a higher and statistically significant (Fisher's exact test: 0.0016) positivity for Listeria spp. than those sold by the shops, independently of the type of cheese. One particular type of cheese proved to be frequently contaminated (Fis. ex. test: 0.0013). Technical, analytical and epidemiological aspects are discussed. PMID- 9353546 TI - Effect of heat on the survival of infectious coxsackievirus B3 and its genome in water. AB - The aim of this study was to estimate the resistance of the viral genome of coxsackie B3 to heat (from 25 degrees C to 95 degrees C) with respect to the infectious virus. The results show that viral genomes were much more resistant to heat than infectious virus. The infectious coxsackie B3 was undetectable following incubation at 55 degrees C for 15 min., while the viral genome was still detected after incubation at 95 degrees C for 15 min. The viral genome became undetectable by RT semi-nested PCR after incubation at 95 degrees C for 30 min. We suggest that the rapid loss of infectious ability of the virus after incubation at 55 degrees C may be mostly due to cleavage or conformation changes in the viral capsid without significant destruction of the genome. PMID- 9353547 TI - An evaluation of certain Salmonella detection methods in surface water. AB - Two different procedures were employed for detecting Salmonella spp. in environmental water samples: a rapid method (enrichment in Salmosyst Broth and plating in Rambach Agar-Merck) and a longer assay (pre-enrichment in Buffered Peptone Water, enrichment in Selenite-Cistyne Broth and plating in Brilliant Green Agar-Difco). The efficiency of microbiological tests was measured by the following criteria: recovery, sensitivity and specificity. The results analysed by the Kendall concordance coefficients demonstrated that the rapid method appeared more effective even if poorly specific. PMID- 9353548 TI - Hospital hygiene in Great Britain. AB - ICT's in the UK are experienced, well trained and are enthusiastic. However, their efforts are frustrated through lack of resources. Infection Control is now a quality issue and defined separate budgets are being established and hospital contracts now contain elements of infection control as part of the service. Infection control is coming of age in the UK after 25 years of earnest effort. PMID- 9353549 TI - The situation of hospital hygiene in Scandinavia. AB - The situation of hospital hygiene in Scandinavian countries, especially Finland, is described. The infection control activities are most often also quality assurance and management, although the infection control staff does not always realize it. The features of infection control in all Scandinavian countries are similar, but they differ in details. Most hospitals have Infection Control Committees with an advisory and expert role in infection control, but normally with no executive power. All Scandinavian countries have a network of skillful Infection Control Nurses working together with the Infection Control Officer of the hospital. The Scandinavian countries have advanced guidelines for various procedures and other activities that need infection control alertness. The voluntary infection control societies have a major role in the development of infection control activities and education. The development of suitable process and outcome quality indicators are a demanding task for the future. PMID- 9353550 TI - Hospital hygiene in Europe. The situation in Belgium and The Netherlands. AB - The situation of hospital hygiene in Belgium and in the Netherlands is described in the light of the official regulations, the composition and the functioning of the infection committee and of the hospital hygiene team, and the availability of official or semi-official guidelines. Typical for the Netherlands in the long tradition of issuing guidelines on hospital hygiene, in the beginning in 1966 in the form of an advice of the Health Council, at present in the guidelines of the Working Group on Infection Prevention (WIP). A particularity for Belgium is the financing by the state of the hospital hygiene doctor and the hospital hygiene nurses based on a system of scores in which the beds of specialisms with a higher infection risk count for more than general beds. PMID- 9353551 TI - Technical design and assessment of tube equipment using two-phase flow for cleaning and disinfection. AB - Most pipeline systems in dairy and food processing plants are cleaned by circulating cleaning solutions under pressure with a liquid pump. The flow of the circulated solutions is single-phase or flooded flow. Milking system pipelines are subject to special requirements which distinguish them from those in dairy and other food processing plants. Milking system pipelines are considerably larger in diameter than product lines in dairy plants because they must carry both milk and air in a stratified flow condition during the milking process. Milking machine Clean-In-Place (CIP) systems have historically used flooded flow to circulate cleaning solutions. The force to move liquid, however, is typically the vacuum provided by the same vacuum pump used during milking, rather than a positive pressure liquid pump. As the size and complexity of milking machines has increased in recent years, flooded flow CIP systems have become inadequate. The amount of water required to fully flood a milking system becomes impractical with very long and/or large diameter pipelines. The power available to achieve adequate flow velocity is also limited. Air admission has been used to produce two-phase (air/water) slug flow and overcome some of the limitations of fully flooded CIP. Cycled air admission can reduce the amount of water required for circulation and increase flow velocities and thus enhance mechanical cleaning action. Cycled air admission has been implemented in the field largely through trial and error methods. There has been a lack of fundamental design information and testing protocols for air-injected milking machine CIP systems. This has resulted in mixed success in the application of air injected systems. This paper summarizes both laboratory and field research conducted at the University of Wisconsin Milking Research and Instruction lab to provide basic information for the design of air injected CIP systems and methods for field assessment of these systems. Just as properly implemented air-injection can improve cleaning and sanitation in milking machines with less water and energy, so to it may have applications in the cleaning of other types of tube equipment. PMID- 9353552 TI - In vitro assessment of asbestos fibers genotoxicity. AB - This study was carried out in order to assess the genotoxic effect of in vitro exposure to commercial chrysotile asbestos. V 79 cell line, known as a well established cellular model, was used for detection of asbestos genotoxic potency. Conventional structural chromosomal aberration analysis and sister chromatid exchange (SCE) method were both used for asbestos genotoxicity assessment. Within the experimental protocol applied, V 79 cells were treated with asbestos in concentrations of 100 and 200 micrograms/ml F-10 (HAM) media during 90 days, respectively. Analysis of changes in chromosome structure as well as of cell ploidy was performed each tenth day of the experimental course, consecutively. Two hundred well spread metaphases were taken into account for chromosomal aberration analysis. Frequency of sister chromatid exchanges was observed in 50 cells per sample. The results of cytogenetic tests revealed structural chromosomal damages, SCE-elevation and changes in cell ploidy. Cytogenetic effect of asbestos obviously depended on the dose applied and on the period of incubation. The results of this study suggest that significant cytogenetic changes occurring after asbestos treatment might directly or indirectly be the part of the biological events responsible for eliciting asbestos-induced carcinogenesis. PMID- 9353553 TI - Definition, prevalence and development of nasal obstruction. PMID- 9353554 TI - Microvascular anatomy of the nose. AB - The microvasculature of the nose consists of: 1) A dense subepithelial network of capillaries, with fenestrations between the endothelial cells. This network provides nutrients to the epithelium and glands, and allows passage of water into the lumen for evaporation and air-conditioning. 2) A system of capacitance vessels or sinuses, which when they distend, block the nasal lumen, and when they empty, open the nasal passages. Changes in their volume will affect the filtering and air-conditioning functions of the nose. 3) Arteriovenous anastomoses which allow rapid passage of blood through the mucosa. They are probably important in air-conditioning, and in the countercurrent mechanisms that tend to keep the brain cool in a hot dry climate. The anatomical interrelationships between these different systems is not well understood, nor is their differential control in terms of actions of mediators and nerves. In neurogenic inflammation sensory nerves are excited and release local mediators such as substance P via axon reflexes. These sensory neuropeptides will cause vasodilatation, vascular congestion and extravasation of liquid from the postcapillary venules, with resultant oedema and exudate. They may also cause secretion from the submucosal glands. PMID- 9353555 TI - Mediators of nasal blockage in allergic rhinitis. PMID- 9353556 TI - Measurement of nasal patency. PMID- 9353557 TI - Nasal allergen challenge: changes in nasal patency and in biochemical markers. PMID- 9353558 TI - Rhinitis medicamentosa: aspects of pathophysiology and treatment. AB - With modern vasoconstrictors, such as oxy- and xylometazoline, the risk of developing rhinitis medicamentosa (RM) has been considered to be small or even nonexistent. However, recent studies have shown that overuse of these drugs may result in rebound congestion, nasal hyperreactivity, tolerance, and histologic changes of the nasal mucosa. Using rhinostereometry, it has also been shown that the long-term use of the preservative benzalkonium chloride (BKC) in oxymetazoline nasal spray accentuates the severity of rhinitis medicamentosa in healthy volunteers. A nasal decongestant spray composed of a combination of vasoactive substances and BKC has a long-term adverse effect on the nasal mucosa. BKC alone induces mucosal swelling after 30 days use of the nasal spray in healthy subjects, unlike placebo. According to the author, rhinitis medicamentosa can be defined as a condition of nasal hyperreactivity, mucosal swelling, and tolerance that is induced, or aggravated, by the overuse of topical vasoconstrictors with or without a preservative. An adequate treatment of these patients consists of a combination of vasoconstrictor withdrawal and a topical corticosteroid to alleviate the withdrawal process. The underlying nasal disorder must then be treated. Patients with rhinitis medicamentosa who overuse topical decongestants and are able to stop using such drugs should be careful about taking these drugs again, even for a few days. They must be informed about the rapid onset of rebound congestion upon repeated use in order to avoid the return of the vicious circle of nose-drop abuse. PMID- 9353559 TI - Antihistamines and nasal blockage. PMID- 9353560 TI - Effect of corticosteroids on nasal blockage in rhinitis measured by objective methods. AB - This paper gives an overview of placebo-controlled studies of the effect of corticosteroid treatment on nasal blockage, based on objective measurements of nasal airway patency. A few studies of perennial rhinitis have indicated that pretreatment with an intranasal corticosteroid has a moderate effect on nasal hyperresponsiveness, measured as the histamine-induced increase of nasal blockage. Whereas the effect on allergen-induced early-phase symptoms is variable, the effect on the late-phase blockage is almost complete. In seasonal allergic rhinitis, a few studies have shown an effect of intranasal steroids on nasal airway resistance, nasal peak flow and on acoustic rhinometry, but there are no reports on the effect in adults with perennial rhinitis. In children with perennial disease, intranasal treatment results in increased nasal patency and, in one study, also in reduced mouth breathing and in an increased threshold for exercise-induced bronchoconstriction. In patients with nasal polyposis, intranasal steroids have an effect on nasal airway resistance and on nasal peak flow both before and after polypectomy. There is convincing evidence that intranasal corticosteroids provide a better effect than antihistamine on nasal blockage. Amazingly, there does not appear to be any report on the effect of systemic corticosteroid treatment on nasal airway patency, and it is therefore difficult to recommend this treatment in a rational dosage. In conclusion, there is a fairly good documentation in support of the efficacy of intranasal steroid treatment on nasal airway patency in rhinitis. An objective measurement of nasal airway patency ought to be the routine in controlled rhinitis trials. PMID- 9353561 TI - Nasal blockage in children with non-infectious rhinitis: consequences and treatment. PMID- 9353562 TI - Cloning and expression of the cDNA encoding prolyl oligopeptidase (prolyl endopeptidase) from bovine brain. AB - Prolyl oligopeptidase (EC 3.4.21.26, prolyl endopeptidase) cDNA from bovine brain was cloned by PCR, and the amplified fragment was used as a probe to screen the cDNA library from bovine brain. The obtained clone contained a 2.7 kb DNA fragment with an open reading frame of 2130 nucleotides, and encoded a protein of 710 amino acids with a deduced molecular weight of 80640 Da. The deduced amino acid sequence is 95, 94, 51 and 48% homologous to those of human T-cell, porcine brain, Aeromonas hydrophila, and Flavobacterium meningosepticum prolyl endopeptidases, respectively. The bovine brain prolyl endopeptidase-encoding cDNA was expressed using an expression vector bearing a tac promoter, with an approximate yield of 20 micrograms/ml of cell culture. PMID- 9353563 TI - Structure-related pharmacokinetics of xanthines after direct administration into the peritoneal cavity of rats. AB - The pharmacokinetic characteristics, peritoneal permeability and hydrophobicity of three xanthine derivatives, theophylline, enprofylline and 1-methyl-3 propylxanthine (MPX), were investigated in rats. Isotonic saline (30 ml) containing xanthine (2.5, 5 and 10 mg/kg) and blue dextran (0.2%) was administered intraperitoneally. The pharmacokinetic parameters of these xanthines were estimated using concentration-time data obtained from the peritoneal cavity and systemic circulation. Disappearance of these xanthines from the peritoneum declined in almost a monoexponential manner regardless of the dose administered. The volume of distribution (33.9 ml) in the peritoneal cavity was similar to the injection volume, indicating that dialysate was not diluted by the fluid in the peritoneal cavity and the effect of drug adsorption on the peritoneal membrane was minimal. The pharmacokinetics of MPX was dose-dependent, but that of theophylline and enprofylline was not. The fraction of the administered dose absorbed through the peritoneal cavity was 0.71, 0.85, 0.93 for theophylline, enprofylline and MPX, respectively. The peritoneal clearance was significantly different (p < 0.05) among the three xanthines by two-way analysis of variance, and a strong correlation was noted between their peritoneal clearance and hydrophobicity (r = 0.98, p < 0.01). These findings suggest that hydrophobicity is an important determinant in the peritoneal permeation of these xanthines. PMID- 9353564 TI - Inhibitory effect of a new ureidophenol derivative T-2591 on LDL oxidation and ACAT activity. AB - We investigated the effects of T-2591, a new ureidophenol derivative, on low density lipoprotein (LDL) oxidation, acyl CoA: cholesterol acyltransferase (ACAT) and foam cell formation of macrophages in vitro. T-2591 inhibited both copper ion -and endothelial cell--induced LDL oxidation with higher potencies than probucol did. It inhibited ACAT from rabbit intestine, liver and aorta, the respective IC50 values being 0.26, 4.6 and 4.1 microM. It also inhibited ACAT from the mouse macrophage cell line J774 A.1, and its IC50 value (0.067 microM) was much lower than that of CI-976 (4.1 microM). This probably accounts for the inhibition of foam cell formation measured as cholesteryl ester formation in both mouse peritoneal macrophages and J774 A.1 cells at low concentrations (IC50; 0.06 and 0.44 microM, respectively). These observations suggest that T-2591 should be evaluated as a potential tool to retard atherosclerosis in animal models. PMID- 9353565 TI - Antitumor effects and toxicities of carboxymethylpullulan-peptide-doxorubicin conjugates. AB - In vivo antitumor effects of the conjugates of doxorubicin (DXR) with carboxymethylpullulan (CMPul) through tetrapeptide spacers were compared with those of DXR against tumor-bearing rats. CMPul-DXR conjugates bound through Gly Gly-Phe-Gly and Gly-Phe-Gly-Gly spacers were found to be more potent than DXR after a single intravenous injection in rats bearing Walker 256 carcinosarcoma. These conjugates were also more effective than DXR in rats bearing Yoshida sarcoma. However, CMPul-DXR conjugate bound through Gly-Gly-Gly-Gly was less effective against Walker 256-bearing rats than DXR. Body weight loss of CMPul-DXR conjugates in rats, on the other hand, was less than that of DXR at a DXR dose of 10 mg/kg. Lethal doses of CMPul-DXR conjugates in CDF1 mice were about 3-times higher than that of DXR. These data suggest that the therapeutic index of CMPul DXR conjugates bound through appropriate peptide spacers was increased more than that of DXR. However, CMPul-DXR conjugates tested were all less effective than DXR against Walker 256 cells in vitro. Also, 125I-labeled CMPul-DXR conjugate accumulated much less in the cells than 14C-DXR. PMID- 9353566 TI - Roles of gamma-aminobutyric acidB (GABA B) and gamma-hydroxybutyric acid receptors in hippocampal long-term potentiation and pathogenesis of absence seizures. AB - Experiments were performed to examine the roles of gamma-aminobutyric acid(B) (GABA(B)) and gamma-hydroxybutyric acid (GHB) receptors in long-term potentiation (LTP) of the hippocampal CA1 region in vivo and in the genesis of the spike and wave discharges (SWDs) associated with absence seizures. When tetanic stimulation was delivered to the CA3 region, stable LTP was observed in the CA1 region in saline-treated mice. In mice treated with 5 mg/kg baclofen, the population spike amplitude was significantly potentiated by tetanic stimulation and the degree of potentiation was the same as that induced in saline controls. However, this potentiation decayed to the baseline level about 90 min after stimulation. The decay was reversed by pretreatment with 200 mg/kg P-[3-aminopropyl]-P diethoxymethylphosphinic acid (CGP 35348), a selective GABA(B) receptor antagonist. In mice treated with 50 mg/kg gamma-butyrolactone (GBL), a prodrug of GHB, stable LTP was observed 90 min after tetanic stimulation and was greater than that in saline controls. GBL-induced potentiation of LTP was antagonized by 50 mg/kg NCS 382, a putative GHB receptor antagonist. Administration of baclofen (20 mg/kg) or GBL (70 mg/kg) induced absence-like seizures associated with 3-6 Hz SWDs, and CGP 35348 suppressed both baclofen- and GBL-induced SWDs. NCS 382 also attenuated SWDs induced by GBL and baclofen. These results suggest that baclofen and GHB have different effects on LTP in the CA1 region of the hippocampus in vivo, although they have a common mode of action on the thalamocortical functions related to the pathogenesis of absence seizures. PMID- 9353567 TI - Immunosuppressive properties of MX-68, a novel unpolyglutamatable antifolate. AB - MX-68 is a novel unpolyglutamatable antifolate. We here reported the in vitro and in vivo immunosuppressive properties of MX-68 compared with a polyglutamatable antifolate, methotrexate (MTX). MX-68 showed potent suppressive effects on mitogen-induced mouse splenic lymphocyte proliferation as well as immunoglobulin production from LPS-stimulated mouse splenic B cells. In in vivo studies, MX-68 significantly suppressed antigen-specific antibody production of both T cell dependent antigen and T cell-independent antigen. Moreover, MX-68 inhibited a sheep red blood cell (SRBC)-induced delayed-type hypersensitivity (DTH) reaction by administration starting from the day of antigen immunization, but did not suppress the effector phase of the DTH reaction. MTX showed suppressive activities similar to MX-68 in all experiments. Interestingly, although MX-68 demonstrated somewhat stronger suppressive effects than MTX in vivo, the results from in vitro studies were reversed. These results suggest that polyglutamation is not always required to suppress immune responses and that MX-68 is a slightly stronger immunosuppressive drug than MTX in mice. PMID- 9353568 TI - Anti-platelet aggregation activity of some pyrazines. AB - This report describes the anti-platelet aggregation activity of 48 pyrazines. Among alkyl- and arylpyrazines tested, 2,3-diphenylpyrazines showed the strongest anti-platelet aggregation activity. Then, various substituents were introduced into the phenyl groups, and the 2,3-bis(p-methoxyphenyl)pyrazine derivatives were consequently found to possess considerably strong inhibitory activity. PMID- 9353569 TI - Enhancement of in vivo anti-influenza virus activity of 5,7,4'-trihydroxy-8 methoxyflavone by drug delivery system using hydroxypropyl cellulose. AB - Enhancement of in vivo antiviral activity of 5,7,4'-trihydroxy-8-methoxyflavone (F36) against H3N2 subtype of influenza A virus by drug delivery system (DDS) with hydroxypropyl cellulose (HPC) was studied. Although in the absence of HPC F36 (0.5 mg/kg) showed no antiviral activity against mouse-adapted influenza virus A/Guizhou/54/89 (H3N2) in mice, when F36 solution containing HPC was administered intranasally 5 min after the virus inoculation, proliferation of the virus in both nasal and broncho-alveolar cavities was inhibited significantly. The relationship between concentration (0.2-0.5%) and deposition ratio of HPC was studied. When 10 microliters of fluorescein isothiocyanate (FITC)-conjugated HPC solution was administered intranasally to BALB/c mice, deposition ratio of HPC at 6 h after inoculation in nasal cavity was dependent on its concentration. The deposition ratio of HPC in broncho-alveolar cavity, however, was reversely dependent on its concentration. Anti-influenza virus activity of F36 in nasal and broncho-alveolar cavities was dependent both on the concentration and deposition ratio of HPC. HPC was most effective at 0.5% in nasal cavity and at 0.3% in broncho-alveolar cavity. These results indicate that DDS with HPC enhances the anti-influenza virus activity of F36 in vivo. PMID- 9353570 TI - Studies on kochiae fructus. III. Antinociceptive and antiinflammatory effects of 70% ethanol extract and its component, momordin Ic from dried fruits of Kochia scoparia L. AB - The 70% ethanol extract (KS-ext) from Kochiae Fructus (dried fruits of Kochia scoparia L.) was screened for its activity on nociceptive and inflammatory responses in experimental animals. Although KS-ext at an oral administration of 500 mg/kg had an antinociceptive effect on writhing responses induced by acetic acid, it was ineffective on nociceptive response in the hot plate test. Oleanolic acid oligoglycoside, momordin Ic isolated from Kochiae Fructus significantly decreased the frequency of licking behavior within a unit of time at the late phase without affecting that of the early phase in the formalin test. Also, KS ext inhibited the rise of vascular permeability induced by acetic acid, the increase of paw edema induced by carrageenin, histamine, serotonin or bradykinin and ear swelling induced by arachidonic acid. Momordin Ic also exhibited an inhibitory effect on carrageenin-induced edema. These results indicated that Kochiae Fructus has a peripheral antinociceptive effect mediated by antiinflammatory action, and that its active component can be partially attributed to momordin Ic. PMID- 9353571 TI - Effects of escins Ia, Ib, IIa, and IIb from horse chestnut, the seeds of Aesculus hippocastanum L., on acute inflammation in animals. AB - We investigated the effects of escins Ia, Ib, and IIb isolated from horse chestnut, the seeds of Aesculus hippocastanum L., and desacylescins I and II obtained by alkaline hydrolysis of escins on acute inflammation in animals (p.o.). Escins Ia, Ib, IIa, and IIb (50-200 mg/kg) inhibited the increase of vascular permeability induced by both acetic acid in mice and histamine in rats. Escins Ib, IIa, and IIb (50-200 mg/kg) also inhibited that induced by serotonin in rats, but escin Ia didn't. Escins Ia, Ib, IIa, and IIb (200 mg/kg) inhibited the hind paw edema induced by carrageenin at the first phase in rats. Escin Ia (200 mg/kg) and escins Ib, IIa, and IIb (50-200 mg/kg) inhibited the scratching behavior induced by compound 48/80 in mice, but escin Ia was weakest. Desacylescins I and II (200 mg/kg) showed no effect. With regard to the relationship between their chemical structures and activities, the acyl groups in escins were essential. Escins Ib, IIa, and IIb with either the 21-angeloyl group or the 2'-O-xylopyranosyl moiety showed more potent activities than escin Ia which had both the 21-tigloyl group and the 2'-O-glucopyranosyl moiety. PMID- 9353572 TI - Biological properties of conjugates of mitomycin C with estradiol benzoate and estradiol: their stability characteristics in biological media and their binding abilities to estrogen receptor. AB - Conjugates of mitomycin C (MMC) with estradiol benzoate and estradiol via glutaric acid, abbreviated to EB-glu-MMC and E-glu-MMC, respectively, were investigated in vitro to determine their stability and MMC regeneration properties in biological media and on their binding to estrogen receptor. EB-glu MMC and E-glu-MMC were added into a mixture of 1/15 M phosphate buffer, pH 7.4 (ionic strength = 0.3), propylene glycol (PG) and rat plasma (4:5:1, v/v/v), named 10% plasma, or into a mixture of the buffer, PG and rat liver homogenate (9:10:1, v/v/w), named 5% liver homogenate, and each was incubated at 37 degrees C. The conversion characteristics of EB-glu-MMC and E-glu-MMC were compared with those in the buffer-PG (1:1, v/v) mixture previously reported. In 10% plasma, the change of EB-glu-MMC to E-glu-MMC was accelerated enzymatically to some extent, while the enzymatic degradation of E-glu-MMC was not accelerated at all. In 5% liver homogenate, EB-glu-MMC changed quickly to E-glu-MMC, whereas the degradation of E-glu-MMC was accelerated very little. E-glu-MMC was considered to be rather stable against enzyme in the biological media. Competitive binding studies using the rat uterine estrogen receptor showed that the specific binding affinity of E-glu-MMC was 0.81% to that of estradiol, while EB-glu-MMC hardly exhibited specific binding. E-glu-MMC was regarded as a hormone-drug conjugate showing a small specific binding affinity to the estrogen receptor. E-glu-MMC is considered to be an effective antitumor agent which gradually generates MMC in the body, and its receptor-mediated action to target cells such as estrogen receptor-positive tumor cells might be possible. PMID- 9353573 TI - Inactivation of a particle beta-glucan by proteins in plasma and serum. AB - (1-->3)-beta-D-Glucans remained in the liver and spleen for long time, i.e. more than a month, without major structural changes/because there is no specific metabolic pathway for it in the body. However, biological activities, such as priming activity to LPS, triggered TNF-alpha synthesis, and antitumor activity was reduced more quickly. In this paper, we demonstrated the contribution of protein binding in inactivating beta-glucans. A particle beta-glucan preparation, zymosan, was treated with serum or plasma at 37 degrees C and their various biological activities were compared with zymosan alone. Such biological activities as antitumor activity, TNF-production, IL-6 production, complement activation and vascular permeability were significantly decreased by serum or plasma treatment. These results strongly suggested that the binding of serum or plasma protein(s) to beta-glucans would be a key step in inactivating a particle beta-glucan in the body. PMID- 9353575 TI - Influence of diet on the single-dose pharmacokinetics of isosorbide 5-mononitrate and sustained-release isosorbide dinitrate. AB - The influence of diet on the single-dose pharmacokinetics of isosorbide 5 mononitrate (IS-5-MN) and sustained-release isosorbide dinitrate (ISDN) was studied in 8 healthy male volunteers. The subjects received either a low calorie/low-fat diet (LCFD) or a high-calorie/high-fat diet (HCFD) according to a cross-over schedule and were administered the drug in tablet form after breakfast. The study was conducted first in 8 subjects who received a 20 mg IS-5 MN tablet and then in 6 of these individuals who received a 20 mg sustained release ISDN tablet. After oral doses of IS-5-MN, the plasma drug levels declined monoexponentially and could be described by a one-compartment open model. There were no significant differences in the pharmacokinetic parameters for IS-5-MN between the LCFD and the HCFD. After administration of sustained-release ISDN, the plasma ISDN levels showed marked variations both between and within individuals. The increase in the mean AUC0-12 values for ISDN with the HCFD was found to exceed 60%, although there was no significant difference between the two diets. PMID- 9353574 TI - The novel compound NO-1886 activates lipoprotein lipase in primary cultured adipose and skeletal muscle cells. AB - As previously reported, we have discovered that a novel compound, NO-1886 (diethyl 4-[(4-bromo-2-cyanophenyl)carbamoyl] benzylphosphonate) has a powerful lipoprotein lipase (LPL) stimulating activity. Oral administration of NO-1886 increased LPL activity in postheparin plasma of experimental animals, resulting in the reduction of plasma triglyceride with concomitant elevation of high density lipoprotein cholesterol. However, the mechanism of NO-1886 on LPL activity is not clearly understood. To address this problem, we examined the effect of NO-1886 on LPL activity in primary rat cell culture isolated from adipose and skeletal muscle tissue. NO-1886 increased total LPL activity 18% and 23% in adipocytes at a dose of 3 and 10 micrograms/ml, respectively, and 43% at a dose of 10 micrograms/ml in skeletal muscle cells. These results indicate that NO 1886 may act directly on LPL-producing cells such as adipose and skeletal muscle. PMID- 9353576 TI - Absorption of oligodeoxynucleotide by suppository from rat rectal route. AB - Rectal absorption of 32-mer phosphorothioate deoxynucleotides (S-Oligo) in the rat was attempted with the aid of a suppository containing oleic acid and a surfactant. Although, the suppository without adjuvant did not show detectable blood levels, that containing over 10% oleic acid enabled the absorption of S Oligo with tmax at 2 h postdosing. PMID- 9353577 TI - Remarkable inhibitory effects of hybrid liposomes on the growth of HL-60 cells coupled to induction of apoptosis. AB - The hybrid liposomes (90 mol% DMPC/10 mol% C12(EO)10 or C12(EO)12) have a highly inhibitory effect on the growth of tumor cells (HL-60). The induction of apoptosis by the hybrid liposomes in HL-60 cells was revealed on the basis of flow cytometry and DNA electrophoresis. PMID- 9353578 TI - Three-year coronal caries incidence in older Canadian adults. AB - This paper describes the incidence of coronal caries in a sample of older adults. A 3-year follow-up study was conducted of 493 community-dwelling adults aged 50 years and over in Ontario, Canada. The incidence of coronal caries was 57.0%, and the mean net DFS increment was 1.9 surfaces. In bivariate analysis, several variables were significantly associated with incidence and/or mean DFS increment. These included: age, marital status, baseline coronal DFS, number of teeth at baseline, mean periodontal attachment loss of 4 mm or more, and wearing partial dentures. In logistic regression analysis only four factors had significant independent effects. These were level of education, marital status, mean periodontal attachment loss and number of teeth at baseline. The predictive ability of this model was fair: accuracy 65.7%, sensitivity 80.2%, and specificity 46.2%. When logistic analysis was repeated separately for two age groups, different predictors had significant independent effects, and sensitivity and specificity values differed substantially. These findings indicate predictive models for caries incidence should include both clinical and non-clinical variables because both types of variables may help to explain different aspects of coronal caries experience. Further research is required to identify other factors associated with coronal caries in older adults. PMID- 9353579 TI - A clinical and microbiological study of deep carious lesions during stepwise excavation using long treatment intervals. AB - Concern about the survival of microorganisms in deep carious lesions may often lead to unnecessary exposure of the pulp during final excavation. There are reasons, therefore, to initiate systematic studies on the alternative procedure known as stepwise excavation. Clinical evaluation of stepwise excavation was performed on 31 deep carious lesions considered to result in pulp perforation by traditional excavation. This study examines the clinical and microbiological alterations during the final excavation performed during long intervals (6-12 months) after the initial treatment that included peripheral dentine excavation and removal of the central cariogenic biomass and the superficial necrotic dentine. The dentine colour and consistency were assessed by means of standardized scales before application of a Ca(OH)2 compound and a temporary sealing for 6-12 months. Reassessments were performed before the after final excavation. Microbiological dentine samples were obtained in 19 randomly selected lesions by a sterile bur, transferred to and diluted in reduced transport fluid, and plated on tryptic soy agar. After anaerobic incubation at 37 degrees C for 7 days, total colony-forming units per millilitre were counted from (1) peripheral excavated and hard dentine (control), (2) central demineralized dentine before and final excavation, and (3) central dentine after the final excavation. Six samples of central demineralized dentine were without any cultivable flora increasing to 9 samples after the final excavation. The clinical dentine changes occurring during stepwise excavation were characterized by enhanced hardness of the dentine which was associated with a marked reduction in bacterial growth after the final excavation. Despite the presence of bacteria in the excavated dentine none of the carious lesions resulted in pulp perforation, suggesting that the initial removal of the cariogenic biomass appears to be essential for control of caries progression. Stepwise excavation is not only an appropriate treatment of deep carious lesions but is also considered a suitable model for microbiological studies to determine the bacteria persisting in clinically excavated lesions. PMID- 9353580 TI - An in vitro comparison of three fluoride regimens on enamel remineralization. AB - The purpose of this study was to compare the enamel remineralization effectiveness of a fluoride rinse, fluoridated dentifrice, and fluoride-releasing restorative material. Forty extracted molars had 1 x 5 mm artificial carious lesions formed at the interproximal contact point. One-hundred-micrometer sections were obtained at the caries sites, and polarized light photomicrographs were obtained. The sections had varnish placed, leaving only the external section site exposed, and were situated back into the original tooth. Forty other molars were obtained; 10 had Class-II glass ionomer cement restorations placed. These 40 teeth were mounted to have interproximal contact with the adjacent teeth containing artificial carious lesions. Specimens were placed in closed environments of artificial saliva for 1 month, with saliva being changed every 48 h. Ten specimen pairs were brushed with a fluoridated dentifrice for 2 min, twice per day, 10 specimen pairs were rinsed with a 0.05% sodium fluoride rinse for 1 min twice per day, 10 specimen pairs had Class-II glass ionomer cement restorations positioned adjacent to 10 teeth with artificial carious lesions, and 10 specimen pairs acted as controls. After 30 days, the same sections were photographed again under polarized light, and areas of the lesions were digitized quantitatively. Results demonstrated the mean (+/- SD) remineralization (mu m2) in Thoulet's 1.41 imbibition media to be: lesions adjacent to glass ionomer cement restorations, 2.45 +/- 170; lesions exposed to a fluoridated dentifrice, 223 +/- 102; lesions exposed to 0.05% sodium fluoride rinse, 374 +/- 120, and control lesions only exposed to artificial saliva, 101 +/- 69. Duncan's analysis indicated the fluoridated rinse to have significantly greater remineralization effects on adjacent caries than the other groups (p < or = 0.05). The glass ionomer restorative material and fluoridated dentifrice also had significantly greater remineralization effects on adjacent caries than the control, yet significantly less than the fluoridated rinse (p < or = 0.05). PMID- 9353581 TI - Remineralization of root surfaces demineralized in solutions of differing fluoride levels. AB - The beneficial effects of fluoride on enamel have been well documented. However, limited data are available concerning the amount of fluoride required for beneficial effects on tooth root. Although studies have shown that fluoride inhibits root demineralization, the aim of this study was to investigate the location, extent and amount of remineralization on root dentin substrates after demineralization has occurred. The root surfaces of extracted human teeth were demineralized in a pure chemical buffer containing varying concentrations of sodium fluoride. After this lesion initiation, the same root sections were then placed into a remineralizing solution. The root sections were characterized after demineralization, and again after remineralization, by polarized light microscopy (PLM) and microradiography (MRG). Lesion depths after the demineralization phase were found to be inversely proportional to the fluoride concentration. When fluoride was present, bands or lines within the body of the lesion were observed with PLM and MRG. Using quantitative MRG, variations in mineral content and distribution were recorded. Examination of the root sections after the remineralization phase showed remineralization to have occurred on the remaining mineral and not on organic matrix devoid of mineral. The amount and location of mineral deposition may be of great significance in the arrestment and treatment of in vivo root surface caries. PMID- 9353582 TI - Effectiveness of calcium lactate added to food in reducing intraoral demineralization of enamel. AB - Following the demonstration that rinses with solutions of soluble calcium salts reduced sucrose-induced demineralization, a study was undertaken to determine whether a similar effect could be obtained by the supplementation of a solid food with calcium lactate (CL). Subjects wore palatal appliances containing blocks of bovine enamel that were coated with Streptococcus mutans IB 1600 and ate 5-gram portions of cookies made with defined levels of CL. Determinations were made of changes in iodide penetrability (delta Ip) of the enamel, as well as the pH, calcium and inorganic phosphate of the streptococcal plaque. CL at 3.2% (w/w) reduced delta Ip from 12.9 +/- 1.7 to 6.1 +/- 0.9 units, i.e. by 52.7%. Plaque pH was not affected. Demineralization was reduced progressively with increasing concentrations of added CL, and CL was most effective with increasingly sweet cookies. Plaque contained 32.4 +/- 6.0 and 17.1 +/- 4.2 mM calcium after 1 and 5 min, respectively. Calculations showed that the plaque was saturated with respect to enamel during the first 5-10 minutes after food ingestion, in spite of the progressive drop in plaque pH. In conclusion, the present study demonstrated the reduction of the cariogenic potential of solid food by relatively low concentrations of CL. The effect appeared to be related to the ability of the food to maintain high levels of calcium in the streptococcal plaque during the period of active acidogenesis. PMID- 9353583 TI - Interaction of zinc with a synthetic calcium phosphate mineral. AB - As zinc has been included in several oral health products as an anticalculus and antiplaque agent, the interaction of zinc with a synthetic phosphate was investigated. The synthetic calcium phosphate used in this study was beta tricalcium phosphate, or whitlockite, which is a major constituent of mature calculus. The aim of this work was to study the mechanism of uptake of zinc to this mineral. Zinc was readily taken up by the calcium phosphate to a maximum level of 13.9 mumol/m2. The interaction was reversible and followed a Langmuir adsorption isotherm. There was no concomitant release of calcium with zinc uptake. Inclusion of calcium in the exposure solution did however marginally depress the acquisition of zinc (12% max), but fluoride had no significant effect on uptake. PMID- 9353584 TI - Adsorption of Streptococcus sobrinus dextranase inhibitor to water-insoluble alpha-D-glucans of oral streptococci. AB - A low molecular weight dextranase inhibitor from Streptococcus sobrinus has previously been identified and purified. The range of conditions under which inhibition occurs, and the situations in which dextranase activity of S. sobrinus can reappear, have been examined in the chemostat. These studies have revealed that when dextranase production exceeds that of the inhibitor, all the inhibitor is tightly bound into enzyme-inhibitor complexes, and the excess enzyme remains active. Another factor that influences the activity of dextranase inhibitor has now been identified, namely the ability of the inhibitor to bind to water insoluble glucans. Adsorption to water-insoluble alpha-D-glucans, produced by oral streptococci that were grown in batch culture, increased with their proportion of alpha-1,3-linked sequences of glucose residues. Studies with water insoluble dextrans of Leuconostoc mesenteroides strains showed that alpha-1,6 linked sequences were also important for binding. The inhibitor was not active when adsorbed to glucan, but active inhibitor was released by incubation with soluble dextran. The interactions of sucrose, alpha-D-glucosyltransferases, alpha D-glucans, dextranase and dextranase inhibitor are discussed in relation to the growth rate of S. sobrinus. At low growth rate in the chemostat the predominant alpha-D-glucosyltransferase (GTF) is a GTF-S that converts sucrose into soluble dextran, and the activity of free dextranase inhibitor in the culture filtrate is high. By contrast, at high growth rate the streptococci produce GTFs capable of synthesizing water-insoluble alpha-D-glucans, and no free inhibitor is found in culture filtrate. Thus the activity of free, extracellular dextranase inhibitor is controlled by (i) the extent of binding to dextranase and (ii) the extent of adsorption to water-insoluble alpha-D-glucan. PMID- 9353585 TI - Intra-oral variations in total plaque fluoride related to plaque pH. A study in orthodontic patients. AB - The aim of the present investigation was to study intra-oral variations in total plaque fluoride, and to examine whether such variations were related to plaque pH. Five orthodontic patients abstained from oral hygiene and daily fluoride rinsing for 2 days. Resting and fermenting plaque pH was measured with a touch micro-electrode at 14-21 localized sites on bonded vestibular tooth surfaces in each subject. Plaque samples from the same sites were analysed with a fluoride micro-electrode. A wide range of plaque pH values and fluoride concentrations were observed. In all subjects plaque pH and total fluoride levels were lower at upper than at lower front teeth. A direct log-linear relationship existed between total fluoride and fermenting plaque pH. In 4 of the 5 subjects this relationship was statistically significant (p < 0.05) and quite strong (adjusted R2 0.2-0.5, Beta 0.5-0.7). The study shows significant intra-oral variations in total plaque fluoride related to plaque pH. PMID- 9353586 TI - Protective effect of hexetidine against in vitro bacterial demineralisation of bovine enamel and dentin in the presence of fluoride. AB - Bovine enamel and dentin specimens were overlaid with acidogenic Streptococcus mutans suspensions in agarose. In this model, the minimal demineralisation inhibiting concentrations (MDIC) of hexetidine was determined in the presence of fluoride. A commercially available mouthwash containing 0.1% (2.9 mmol/l) hexetidine was diluted serially and added to the bacterial suspensions together with 0, 5.3, or 26.3 mumol/l fluoride (NaF). After 22 h of incubation at 37 degrees C the bacterial suspensions were removed and assessed for calcium and lactate. The results showed significant inhibitory effects of hexetidine on the demineralisation of the enamel specimens with a MDIC between 15 and 31 mumol/l hexetidine. In the presence of fluoride, approximately fourfold higher concentrations of hexetidine were needed for a significant additional protection of the enamel. No synergistic effect between hexetidine and fluoride was observed. For the demineralisation of the dentin specimens, the MDIC of hexetidine had a value between 31 and 61 mumol/l. At both these concentrations the dentin specimens were relatively less protected in the presence than in the absence of fluoride, and some synergistic effect between hexeditine and fluoride was observed. PMID- 9353587 TI - Effects of antimitotic agents on secretion and detergent extractibility of adrenal nicotinic acetylcholine receptors. AB - 1. Evidence exists that associations of adrenal nicotinic acetylcholine receptors (nAChRs) with the cytoskeleton play an important role in signal transduction pathways by maintaining these receptors in a functional state. These studies were designed to explore this possibility and elucidate the mechanism by which antimitotic agents inhibit activation of adrenal nAChRs. 2. Functional studies demonstrated that vincristine, tubulozole, podophyllotoxin, and demecolcine inhibited nAChR-stimulated catecholamine release noncompetitively and in a concentration-dependent manner, with IC50 values of 3 (1-10), 5 (2-10), 8 (4-15), and 19 (9-39) microM, respectively. 3. Detergent extraction experiments indicated that approximately 36% of adrenal nAChRs were associated with the detergent insoluble cytoskeletal fraction. When chromaffin cells were first treated with antimitotic agents and then detergent solubilized, a significant reduction occurred in the population of adrenal nAChRs associated with the detergent insoluble cytoskeleton. 4. These studies support an association of adrenal nAChRs with microtubules and suggest that the mechanism by which the antimitotic drugs interfere with the signal transduction pathway is by inducing dissociation of nAChRs from the microtubular network. PMID- 9353589 TI - Nitric oxide participates in the stimulatory and neurotoxic action of endothelin on rat striatal dopaminergic neurons. AB - 1. Our method of real-time monitoring of dopamine release from rat striatal slices revealed that endothelin (ET)-3-induced dopamine release was inhibited by NG-methyl-L-arginine (L-NMMA; 1 mM), an inhibitor of nitric oxide (NO) synthase, while NG-methyl-D-arginine (D-NMMA; 1 mM), an inactive isomer of L-NMMA, had no effect. 2. The inhibition of L-NMMA (0.1 mM) became apparent when tissues were pretreated with tetrodotoxin (1 microM) for 30 min and subsequently exposed to ET 3 (4 microM). 3. L-NMMA (0.1 and 1 mM) dose dependently protected against ET-3 triggered hypoxic/hypoglycemic impairment of striatal responses to high K+. 4. Thus, NO may work as a promoter in mediation of the stimulatory and neurotoxic action of ET-3 on the striatal dopaminergic system, presumably by interacting with interneurons in the striatum. PMID- 9353588 TI - Selective chronic sodium or chloride depletion specifically modulates subfornical organ atrial natriuretic peptide receptor number in young rats. AB - 1. We studied the effects of selective chronic sodium depletion of chloride depletion on atrial natriuretic peptide receptor number in the subfornical organ and paraventricular nucleus of young rats. 2. Sodium or chloride depletion decreased plasma levels of atrial natriuretic peptide, increased plasma renin activity, and induced extracellular fluid volume contraction. Chloride depletion induced more significant changes in extracellular fluid volume contraction than sodium depletion. 3. In the subfornical organ, atrial natriuretic peptide receptor number significantly decreased (30%) after sodium depletion, while chloride depletion induced a smaller, not statistically significant decrease. Conversely, atrial natriuretic peptide receptors located in the paraventricular nucleus of young rats were not significantly affected by sodium or chloride depletion. 4. Water deprivation reversed the decrease in atrial natriuretic peptide receptors produced by sodium depletion. Water-deprived sodium-depleted rats actually had higher numbers of atrial natriuretic peptide receptors in the subfornical organ than control rats. These changes were associated with severe extracellular fluid volume contraction and up regulation of brain vasopressin mRNA steady-state levels. Thus, the direction of change in the number of subfornical organ atrial natriuretic peptide receptors was dependent on the degree of extracellular fluid volume contraction. 5. Our results suggest that atrial natriuretic peptide receptors located in the subfornical organ, and not in the paraventricular nucleus, are selectively regulated by sodium depletion and extracellular fluid volume contraction. PMID- 9353590 TI - Binding of aminoalkylindoles to noncannabinoid binding sites in NG108-15 cells. AB - 1. Aminoalkylindoles, typified by WIN 55212-2, bind to G protein-coupled cannabinoid receptors in brain. Although cannabinoids inhibit adenylyl cyclase in NG108-15 neuroblastoma x glioma hybrid cells, cannabinoid receptor binding in these cells has not been described previously. This study compares pharmacological characteristics of [3H]WIN 55212-2 binding sites in rat cerebellar membranes and in NG108-15 membranes. 2. Although the KD of specified [3H]WIN 55212-2 binding was similar in brain and NG108-15 membranes, the Bmax was 10 times lower in NG108-15 than in cerebellar membranes. In both brain and NG108 15 membranes, aminoalkylindole analogues were relatively potent in displacing [3H]WIN 55212-2 binding. However, IC50 values for more traditional cannabinoids were significantly higher in NG108-15 membranes than in brain, e.g., the Ki values for CP55,940 were 1.2 nM in brain and > 5000nM in NG108-15 membranes. Moreover, sodium and GTP-gamma-S decreased [3H]WIN 55212-2 binding in brain but not in NG108-15 membranes. 3. These data suggest that WIN 55212-2 does not label traditional cannabinoid receptors in NG108-15 cells and that these novel aminoalkylindole binding sites are not coupled to G proteins. PMID- 9353591 TI - The cyclin-dependent kinase (cdk) inhibitors, olomoucine and roscovitine, alter the expression of a molluscan circadian pacemaker. AB - 1. In this study, we determined the effects of the cyclin-dependent kinase (cdk) inhibitors, olomoucine and roscovitine, on the circadian rhythm of optic nerve impulse activity recorded from the eye of the marine snail Bulla gouldiana. 2. We found that olomoucine lengthened period and altered circadian phase in a dose dependent manner without appreciably affecting gene transcription or translation. We also found that the more specific cdk inhibitor, roscovitine, was approximately 10-fold more effective in lengthening circadian period, while the inactive analogue, iso-olomoucine, was ineffective. 3. The current results, along with previous results from our laboratory, are consistent with the hypothesis that the biochemical mechanism responsible for generating the ocular circadian rhythm in B. gouldiana is related to the biochemical mechanism that regulates the eukaryotic cell division cycle, i.e., by modulation of the activity of protein kinases belonging to the cdk family. PMID- 9353592 TI - A time-dependent increase in glial fibrillary acidic protein expression and glutamine synthetase activity in long-term subculture of the GL15 glioma cell line. AB - 1. Astrocytes are the most numerous cellular elements in the central nervous tissue, where they play a critical role in physiological and pathological events. The biological signals regulating astrocyte growth and differentiation are relevant for both physiology and pathology, but they are still little understood. 2. Using a poorly differentiated glioma cell line, GL15, we investigated whether, in long-term subculture, this could upregulate the expression of glial fibrillary acidic protein (GFAP), as described in some rodent astrocyte cell lines. Under the same culture conditions, we investigated glutamine synthetase (GS) activity, growth-associated protein (GAP)-43 expression, and expression of several neutrotrophic factors. 3. A dramatic increase in GFAP expression was evidenced by Western blotting during progressive in vitro growth of GL15 cells. GS specific activity was also upregulated in long-term culture. The time spent in vitro by GL15 cells did not affect GAP-43 and neutrophic factor BDNF and NT3 expression as revealed by RT-PCR analysis. 4. Our results suggest that, in GL15, GFAP and GS genes may have common or integrated regulatory mechanisms elicited at the cell confluency which could be relevant for both astrocyte physiology and astrocyte pathology. These mechanisms are not involved in GAP-43 and neutrophic factor BDNF and NT3 expression. PMID- 9353594 TI - Prion protein is necessary for latent learning and long-term memory retention. AB - 1. The cellular prion protein, designated PrPc, is a key molecule in the prion diseases but its physiological function remains unknown. To elucidate whether PrPc plays some role in the central nervous system, we established a line of mice in which the PrP gene had been disrupted and subsequently conducted long-term observations. 2. Performance in latent learning and passive avoidance was evaluated using water-finding and step-through tests, respectively. 3. PrP-/- mice showed impaired performance in the water-finding test, indicating a disturbance in latent learning, at 23 weeks of age. In the step-through test, although the PrP-/- mice showed normal learning ability and short-term memory retention, they evidenced a significant disturbance in long-term memory retention. 4. These results indicate that PrPc is needed for certain types of learning and memory and that the loss of function of this protein may contribute to the pathogenesis of prion diseases. PMID- 9353593 TI - 1H NMR ganglioside ceramide resonance region on the differential diagnosis of low and high malignancy of brain gliomas. AB - 1. The high-resolution 1H NMR (MRS) spectra of human brain tumor homogenates revealed a broad resonance at 5.3-5.4 ppm in glioblastoma multiforme (N = 16) and brain metastases (N = 3). The broad resonance was identified as ceramide, a sphingosine-fatty acid combination portion of ganglioside, indicating an elevated abundance of monounsaturated fatty acids. GLC analysis of gangliosides in the highly malignant glioblastoma multiforme revealed that the elevated monounsaturated fatty acid is oleic acid (C18:1). The resonance at 5.3-5.4 ppm region was not detectable in normal human brain (N = 2), in meningiomas (N = 2), or in low-grade astrocytomas (N = 12). In normal human brain the abundance of monounsaturated fatty acid is minimal. 2. This investigation was made possible because the method of producing homogenate resulted in (i) no loss of lipids during the process and (ii) a well-homogenised sample, with (iii) no loss in chemical integrity. 3. The properties of tumor gangliosides include antigenic specificity and immunosuppressive activity and the ceramide, a sphingosine-fatty acid combination, noticeably influences the ganglioside immunosuppressive activity. 4. The observation of 1H NMR ceramide resonance in high-malignant brain tumors emphasizes the dramatic role of aberant gangliosides and ceramide precursors on the grade of malignancy and invasiveness. 5. Further insight into the specific nature of the ceramide portion of gangliosides in grading the malignancy of brain tumors should be investigated further. PMID- 9353596 TI - Immobilization stress increases mRNA levels of interleukin-1 receptor antagonist in various rat brain regions. AB - 1. Interleukin-1 receptor antagonist (IL-1Ra), as well as the interleukin-1 beta (IL-1 beta) gene response to immobilization stress (IMS), was examined in the rat brain. The reverse transcription-polymerase chain reaction was employed to determine mRNA levels. 3. IL-1 beta and IL-1Ra mRNA levels peaked at approximately 0.5 and 2-4 hr, respectively. The maximum mRNA levels of IL-1 beta were 15-fold higher than pre-IMS levels, whereas those of IL-1Ra were 250-fold higher in the hypothalamus. 3. After the biosynthesis of IL-1 beta has peaked, IL 1Ra may contribute to attenuation of the IL-1 activity which has been enhanced by IMS. PMID- 9353595 TI - Molecular mechanisms underlying forskolin-mediated up-regulation of human dopamine D2L receptors. AB - 1. Human dopamine (DA) D2long (hD2L) receptors, expressed by Ltk- cells, can be up-regulated by treating the cells with forskolin for 16 hr (Johansson and Westlind-Danielsson, 1994). We have examined some of the molecular mechanisms underlying this forskolin-mediated up-regulation. 2. Forskolin (100 microM, 16 hr), but not 1,9-dideoxyforskolin, a forskolin analogue that is unable to activate adenylyl cyclase and raise intracellular cAMP concentrations, up regulates the hD2L receptor population by 43%. The implication of a cAMP dependent increase in the receptor up-regulation was further substantiated by treating the cells with 8-bromo-cAMP or prostaglandin E1 (PGE1). The forskolin mediated rise in receptor number was blocked by cycloheximide or an antisense phosphorothioate oligodeoxynucleotide (ODN) directed toward the hD2L mRNA. KT5720, a specific protein kinase A (PKA) inhibitor, completely blocked the receptor rise, whereas pertussis toxin (PTX) attenuated the increase considerably. Forskolin also produced an increase in the level of the DA hD2short (hD2S) receptor expressed by Ltk- cells. This increase was 2.5-fold higher than that found for the hD2L receptor. 3. The forskolin-mediated hD2L receptor rise is dependent on de novo protein synthesis, a rise in cAMP levels, PKA activation, and, at least partially, PTX-sensitive G proteins. 4. Long-term increases in intracellular cAMP levels may change the sensitivity of a DA receptor expressing cell to DA by increasing D2 receptor density through enhanced cAMP-dependent transcription. PMID- 9353597 TI - The potential for dietary intervention postpartum in women with gestational diabetes. PMID- 9353598 TI - So what's the difference between teenage boys and girls, anyway? PMID- 9353599 TI - Time for law to catch up with life. PMID- 9353600 TI - Multicenter evaluation of the Micral-Test II test strip, an immunologic rapid test for the detection of microalbuminuria. AB - OBJECTIVE: To assess the performance of the Micral-Test II immunologic test strip for the detection of microalbuminuria, a multicenter evaluation in eight European study sites was performed. RESEARCH DESIGN AND METHODS: Using both the Micral Test II test strip and the routine method for the determination of albumin concentration, we investigated 2,228 urine samples from diabetic patients. Additionally, interperson variability, color stability, and possible interfering factors (temperature, pH, leucocyturia, erythrocyturia, and drugs) were tested. RESULTS: For a cutoff concentration of 20 mg/l with respect to the routine methods, a sensitivity of 96.7% and a specificity of 71% were calculated for the Micral-Test II test strip. The negative predictive value was 0.95, and the positive predictive value was 0.78, with a prevalence of positive samples (laboratory method) of 52%. The interperson variability of color interpretation showed 93% concordant readings. The interference study showed an influence of oxytetracycline, leading to higher readings. There was no interference from pH. A sample temperature of < 10 degrees C led to lower readings. In the case of samples with massive leucocyturia and erythrocyturia that may delete the chromatographic process, waiting an additional 1-2 min is needed before reading. CONCLUSIONS: The results of the multicenter evaluation show that the Micral-Test II test strip permits an immediate and reliable semiquantitative determination of low albumin concentrations in urine samples with an almost user-independent color interpretation. PMID- 9353601 TI - The recurrence of gestational diabetes: could dietary differences in fat intake be an explanation? AB - OBJECTIVE: To present the results of a comprehensive dietary review of a group of women with a recurrence of gestational diabetes mellitus (GDM), compared with a group of women with no recurrence of GDM during a subsequent pregnancy. RESEARCH DESIGN AND METHODS: The dietary intake of 14 women with a recurrence of GDM was compared with 21 women with no recurrence of GDM. Women with GDM in one pregnancy have a recurrence rate of only 30-50%. While the reasons for this have not been determined, dietary factors have been considered probable. RESULTS: The women with a recurrence of GDM consumed 38.4 (by diet history) and 41.4% (by food record) of their total energy intake as fats, compared with 34.1 (P < 0.01) and 33.1% (P < 0.001), respectively, for women with no recurrence. The percentage intake of polyunsaturated, monounsaturated, and saturated fatty acids was similar in both groups. There was a proportionate reduction in carbohydrate intake as a percentage of total energy and in fiber intake in grams for the women with a recurrence of GDM. CONCLUSIONS: When the relationship between saturated fat intake and insulin resistance is considered, the possibility exists that dietary modification of fat intake before and during pregnancy may reduce the recurrence rate of GDM. PMID- 9353602 TI - Metabolic effects of alterations in meal frequency in type 2 diabetes. AB - OBJECTIVE: The effects of altering meal frequency on measures of glucose and lipid metabolism in type 2 diabetes were examined by comparing isocaloric dietary regimens in which daily food intake was provided by three or nine meals each day. RESEARCH DESIGN AND METHODS: A total of 13 free-living men and women with type 2 diabetes or persistently impaired glucose tolerance participated in a randomized crossover study in which three- and nine-meal regimes were followed for 4-week periods. Fasting plasma lipid and lipoprotein, glucose and insulin concentrations were measured at weekly intervals and glucose, insulin, and triglyceride responses following a 75-g glucose load at weeks 2 and 4 of each diet period. Dietary intake was also recorded during these weeks. RESULTS: Nutrient intakes and all measures of carbohydrate and lipid metabolism were similar on the three- and nine-meal regimes. CONCLUSIONS: This longer-term study could not confirm the potential benefits of increased meal frequency suggested by comparable 4-week studies in type 2 diabetic individuals and acute experiments in individuals with diabetes. However, as there were no adverse effects of consuming nine meals per day, it would seem appropriate that meal frequency in those with type 2 diabetes should be left to personal choice, provided that energy balance is maintained. PMID- 9353603 TI - Symmetrization of the blood glucose measurement scale and its applications. AB - OBJECTIVE: To introduce a data transformation that enhances the power of blood glucose data analyses. RESEARCH DESIGN AND METHODS: In the standard blood glucose scale, hypoglycemia (blood glucose, < 3.9 mmol/l) and hyperglycemia (blood glucose, > 10 mmol/l) have very different ranges, and euglycemia is not central in the entire blood glucose range (1.1-33.3 mmol/l). Consequently, the scale is not symmetric and its clinical center (blood glucose, 6-7 mmol/l) is distant from its numerical center (blood glucose, 17 mmol/l). As a result, when blood glucose readings are analyzed, the assumptions of many parametric statistics are routinely violated. We propose a logarithmic data transformation that matches the clinical and numerical center of the blood glucose scale, thus making the transformed data symmetric. RESULTS: The transformation normalized 203 out of 205 data samples containing 13,584 blood glucose readings of 127 type 1 diabetic individuals. An example illustrates that the mean and standard deviation based on transformed, rather than on raw, data better described subject's blood glucose distribution. Based on transformed data: 1) the low blood glucose index predicted the occurrence of severe hypoglycemia, while the raw blood glucose data (and glycosylated hemoglobin levels) did not; 2) the high blood glucose index correlated with the subjects' glycosylated hemoglobin (r = 0.63, P < 0.001); and 3) the low plus high blood glucose index was more sensitive than the raw data to a treatment (blood glucose awareness training) designed to reduce the range of blood glucose fluctuations. CONCLUSIONS: Using symmetrized, instead of raw, blood glucose data strengthens the existing data analysis procedures and allows for the development of new statistical techniques. It is proposed that raw blood glucose data should be routinely transformed to a symmetric distribution before using parametric statistics. PMID- 9353604 TI - Factors related to glycemic control in IDDM and insulin-treated NIDDM patients in current practice. A comparison of care policies. SIEMTIC Group. Studio Italiano Epidemiologico Multicentrico su Terapia Insulin e Controllo Metabolico. AB - OBJECTIVE: To evaluate, under routine conditions, the relation between different diabetes care policies and glycemic control through a by-center analysis procedure aimed at reducing some drawbacks of cross-sectional data. RESEARCH DESIGN AND METHODS: A survey on insulin-treated diabetes care management (IDDM and NIDDM) involved 16 Italian randomly selected diabetes outpatient clinics. A total of 2,142 representative patients were investigated. The standardized HbA1c average value of each center was related, by regression models, to some indicators of center care policy (average number of injections, average BMI, proportion of cases with recent fundus oculi examinations, or frequent visits) as well as to patients' average social levels (employment type). Homogeneity in patient admission criteria is assumed among the investigated centers as a basic condition for the procedure validity. Some known imbalance were controlled for both design and analysis. RESULTS: HbA1c showed a univariate inverse relation with daily number of injections in IDDM (P = 0.0009, r2 = 0.56) but not in NIDDM (P = 0.33). It was inversely related to both fundus examination (IDDM P = 0.04; NIDDM P = 0.099) and qualified employment (IDDM P = 0.06; NIDDM P = 0.026). A stepwise regression analysis left in the model insulin injections (P = 0.0002) in IDDM (total r2 = 0.68) and qualified employment (P = 0.016) and fundus examination (P = 0.14) in NIDDM (total r2 = 0.53), after controlling for age, sex, disease duration, insulin therapy starting delay, and insulin dose per kilogram. CONCLUSIONS: These results suggest that the confirmed benefits of a multiple-injection regimen in IDDM cannot be simply extrapolated to NIDDM, where patients' awareness and medical attention to complications proved to be the most important factors in current practice. PMID- 9353605 TI - Prevalence of diabetes and its risk factors in China, 1994. National Diabetes Prevention and Control Cooperative Group. AB - OBJECTIVE: To determine the prevalence of diabetes and impaired glucose tolerance (IGT) and its risk factors in the Chinese population. RESEARCH DESIGN AND METHODS: This study was a population-based cross-sectional study of 224,251 residents aged 25-64 years in 19 provinces and areas, including cities and rural areas of the north, south, east, and middle part of China. RESULTS: Using the 1985 World Health Organization criteria, the prevalence of diabetes and IGT was 2.5 and 3.2%, respectively, in 213,515 subjects aged 25-64 years. Two thirds (70.3%) of the cases had newly recognized diabetes. The prevalence of diabetes in China is about three times higher than it was 10 years ago. On average, subjects with diabetes are older, have higher personal annual incomes, and more often have a family history of diabetes. They also have higher mean BMI, waist-to-hip ratio (WHR), systolic blood pressure, diastolic blood pressure, and a greater prevalence of hypertension. They perform less physical activity and have less education than people with normal oral glucose tolerance test results. Multiple logistic stepwise regression analysis shows that age, BMI (or WHR), family history of diabetes, hypertension, less physical activity, and higher annual income are independent risk factors of NIDDM, and that low education is also an independent risk factor of NIDDM in people with higher personal annual income. CONCLUSIONS: The prevalence of diabetes in China is increasing with economic development and changes from traditional to modernized lifestyle, especially where people had lower level of education and socioeconomic development. Therefore, Chinese people should attempt to retain certain features of their traditional lifestyle (physical activity, healthy food, moderate body weight). Increased knowledge of risk factors for diabetes may help to prevent a further rapid increase in the prevalence of diabetes in China. PMID- 9353606 TI - Insulin response in a triethnic population: effects of sex, ethnic origin, and body fat. Miami Community Health Study. AB - OBJECTIVE: To assess sex and ethnic differences in hyperinsulinemia/insulin resistance and to examine the impact of percent body fat on such differences. RESEARCH DESIGN AND METHODS: A cross-sectional epidemiological study was performed in a normoglycemic population of African-Americans (n = 159), Cuban Americans (n = 128), and non-Hispanic whites (n = 207) who resided in Dade County, Florida, from 1990 to 1995. The insulin area under the curve (AUC) in response to a standard 75-g oral glucose tolerance test (OGTT) was used as an indicator of hyperinsulinemia/insulin resistance. Analysis of covariance was performed to compare sex and ethnic differences in the insulin AUC. Multiple linear regression was used to evaluate the independent correlates of the insulin AUC. RESULTS: After covariate adjustment for percent body fat, men displayed a significantly higher insulin AUC than did women (P < 0.001). African-Americans and Cuban-Americans each had a significantly higher insulin AUC than did non Hispanic white participants (P = 0.01). Alcohol consumption was inversely related to AUC (P = 0.04). CONCLUSIONS: Despite the greater percentage of body fat in women, the insulin AUC was similar in women and men. After adjustment for the sex difference in percent body fat, women displayed a lower insulin AUC than did men, indicating enhanced insulin sensitivity. These differences by sex and ethnicity in insulin resistance are consistent with established differences in heart disease risk (i.e., higher in men and African-Americans) and suggest that hyperinsulinemia/insulin resistance may partly underlie such differences. PMID- 9353607 TI - Gender differences in hospitalizations for IDDM among adolescents in California, 1991. Implications for prevention. AB - OBJECTIVE: Describe gender differences in hospitalizations for IDDM to investigate the need for gender-specific interventions to reduce diabetes-related morbidity. RESEARCH DESIGN AND METHODS: Analyses were based on hospital discharges with any mention of IDDM (n = 2,889) and the subset of these for IDDM as a principal diagnosis (n = 2,270) in California children, ages 0-18 years during 1991. Pregnancy-related hospitalizations were excluded. RESULTS: Females had more diabetes hospitalizations among discharges with any mention of diabetes, among discharges with diabetes as a principal diagnosis, and among discharges with diabetic ketoacidosis as a principal diagnosis. For diabetes as a principal diagnosis, females had 40% more hospitalizations, 44% more repeated hospitalizations, 23% more individuals hospitalized, and significantly higher rates of hospitalizations for ages 10-14 years (50 vs. 38 per 100,000) and for ages 15-18 years (68 vs. 29 per 100,000). Gender differences occurred primarily in adolescents, were independent of complicating conditions at the time of hospitalization, and were observed for diabetic ketoacidosis alone. CONCLUSIONS: Adolescent females had more diabetes hospitalizations than did males. The underlying cause may be biological or behavioral. Management protocols tailored for young women may be required to reduce hospitalizations for IDDM among females. PMID- 9353608 TI - U.K. Prospective Diabetes Study 27. Plasma lipids and lipoproteins at diagnosis of NIDDM by age and sex. AB - OBJECTIVE: To compare fasting plasma lipids and lipoproteins in male and female patients at diagnosis of NIDDM and to examine age and sex differences in lipid concentrations. RESEARCH DESIGN AND METHODS: Cross-sectional study of fasting plasma total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride in 2,139 male and 1,574 female white patients, aged 25-65 years, at diagnosis of NIDDM. RESULTS: At diagnosis of NIDDM, the mean age +/- SD for men was 52 +/- 9 and 53 +/- 9 years for women; BMI was 28.3 +/- 4.9 and 30.8 +/- 6.7 kg/m2, and fasting plasma glucose was 11.6 +/- 3.6 and 12.4 +/- 3.8 mmol/l, respectively. The mean total and LDL cholesterol were higher in female than in male NIDDM patients, 5.8 +/- 1.2 vs. 5.5 +/- 1.1 and 3.9 +/- 1.1 vs. 3.6 +/- 1.0 mmol/l (both P < 0.001), respectively, while triglyceride levels were similar: geometric mean (1 SD interval) for men and women was 1.8 (1.1-3.1) vs. 1.8 (1.1-2.9) mmol/l. HDL cholesterol was higher in female than in male NIDDM patients, 1.09 +/ 0.2 vs. 1.01 +/- 0.24 mmol/l (P < 0.001); the sex differential for HDL cholesterol was 7% in NIDDM patients compared with 22% in the general population. Data analysis by 5-year age bands showed a significant trend toward lower total cholesterol and triglyceride and higher HDL cholesterol in men diagnosed above the age of 50 years. In female NIDDM patients, lipid concentrations increased with age of diagnosis but reached a plateau above the age of 50 years. CONCLUSIONS: The effect of NIDDM, observed at diagnosis, on plasma lipid and lipoprotein levels is more pronounced in women than in men. This may explain in part why the cardiovascular risk is proportionally higher in female patients. PMID- 9353609 TI - Hyperinsulinemia and the development of ST-T electrocardiographic abnormalities. An 11-year follow-up study. AB - OBJECTIVE: It has been suggested that insulin resistance and consequent hyperinsulinemia promote atherosclerosis, but few prospective studies have reported the relationships between hyperinsulinemia and the development of ST-T abnormalities in the 12-lead resting electrocardiogram (ECG) in populations in which atherosclerosis is rare. RESEARCH DESIGN AND METHODS: A total of 304 Japanese men and women, aged 20-69 years, selected for having high blood glucose or more than a trace-positive urine glucose from a population-based health examination in 1981, were followed for 11 years. Of these, 33 died, 1 from myocardial infarction, while 260/271 living were reexamined in 1992. The 237 subjects with a normal ECG at the baseline examination were analyzed. RESULTS: Incident ST-T abnormalities occurred in 13/237 people. Insulin concentrations were positively associated with the development of ST-T abnormalities (relative risk approximately 8, comparing those in the highest versus lowest quartile of insulin values). Adjustment for age, sex, and systolic blood pressure or other risk factors had little effect on this relationship. CONCLUSIONS: Hyperinsulinemia was related to the development of ST-T abnormalities in ECGs in the absence of the development of clinical signs of atherosclerosis, independent of blood pressure and other risk factors in men and women with mild glucose intolerance. PMID- 9353610 TI - Lipoprotein(a) in android obesity and NIDDM. AB - OBJECTIVE: To assess the level of serum lipoprotein(a) [Lp(a)] in nonobese and obese NIDDM subjects with android body distribution. RESEARCH DESIGN AND METHODS: Serum Lp(a) levels were measured in 30 long-standing NIDDM patients (duration of diabetes 12.5 +/- 3 years, mean +/- SD), with 15 of the patients being obese of android distribution (BMI > 30 kg/m2 and waist-to-hip ratio > 0.8). In addition, there were 15 android obese nondiabetic subjects and 10 healthy subjects serving as the control group. RESULTS: All groups of patients in this study (diabetic, obese, and obese diabetic) showed significantly higher levels of Lp(a) than the healthy control group. Lp(a) concentrations were significantly higher in NIDDM patients with android type of obesity than in nondiabetic androids (24.1 +/- 5.6 vs. 14.8 +/- 2.4 mg/dl, P < 0.001). Significantly greater levels of Lp(a) were found in nonobese subjects with diabetes when compared with obese subjects without diabetes (22.3 +/- 4.1 vs. 14.8 +/- 2.4 mg/dl, P < 0.001). Furthermore, Lp(a) serum concentrations were not dependent on the degree of glycemic control (controlled NIDDM 23.6 +/- 5.0 vs. uncontrolled NIDDM 21.4 +/- 2.7 mg/dl, NS), but were much greater in subjects with diabetes complicated by vascular disease (complicated 26.3 +/- 5.0 vs. uncomplicated 20.5 +/- 2.7 mg/dl, P < 0.001). No correlation was found between Lp(a) and other lipid parameters in this study. CONCLUSIONS: Lp(a) levels are significantly elevated in both android-obese and nonobese NIDDM patients regardless of the degree of glycemic control. Lp(a) is an independent risk factor showing greater elevations in those subjects complicated with diabetic vascular diseases. PMID- 9353612 TI - Diabetic peripheral neuropathy: amelioration of pain with transcutaneous electrostimulation. AB - OBJECTIVE: To evaluate the efficacy of transcutaneous electrotherapy for chronic painful peripheral neuropathy in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: Thirty-one patients with symptoms and signs of peripheral neuropathy were randomized to the electrotherapy or sham treatment (control) group. The electrostimulation was given by a portable unit (H-Wave machine) than generated a biphasic, exponentially decaying waveform (pulse width 4 ms, 25-35 V, > or = 2 Hz). Patients treated each of their lower extremities for 30 min daily for 4 weeks at home. Nine patients from the sham-treatment group participated for a second period, during which all of them received the active electrotherapy. Patient's degree of pain and discomfort was graded on a scale of 0 to 5. RESULTS: In the sham-treated group (n = 13), the neuropathic symptoms improved in five (38%) patients, and the pain score declined from 2.92 +/- 0.13 to 2.38 +/- 0.26 (P < 0.04), suggesting a procedure-related placebo effect. In the electrotherapy group (n = 18), symptomatic improvement was seen in 15 (83%) cases, 3 of which were completely asymptomatic; the pain score declined from 3.17 +/- 0.12 to 1.44 +/- 0.25 (P < 0.01) and the posttreatment pain scores were considerably lower (P < 0.03), indicating a substantial treatment effect over and above any placebo influence. Patients in the electrotherapy group reported greater reduction in symptoms (52 +/- 7% vs. 27 +/- 10% in control subjects, P < 0.05) on an analog scale. Moreover, the electrotherapy decreased pain scores (from 3.0 +/- 0.62 to 1.56 +/- 0.32, P < 0.02) in nine patients who had received sham treatment earlier. CONCLUSIONS: A form of transcutaneous electrotherapy ameliorated the pain and discomfort associated with peripheral neuropathy. This novel modality offers a potential non-pharmacological treatment option. PMID- 9353611 TI - Effects of a quick-release form of bromocriptine (Ergoset) on fasting and postprandial plasma glucose, insulin, lipid, and lipoprotein concentrations in obese nondiabetic hyperinsulinemic women. AB - OBJECTIVE: To assess the effect on various aspects of carbohydrate and lipid metabolism of administering a quick-release formulation of bromocriptine (Ergoset) to obese, nondiabetic, hyperinsulinemic women. RESEARCH DESIGN AND METHODS: Hourly concentrations of prolactin, glucose, insulin, free fatty acid (FFA), and triglyceride were measured for 24 h before and after approximately 8 weeks of treatment with Ergoset. In addition, fasting lipid and lipoprotein concentrations and the steady-state plasma glucose (SSPG) concentration in response to a continuous infusion of somatostatin, insulin, and glucose were determined before and after Ergoset administration. RESULTS: Circulating prolactin concentrations were dramatically decreased (P < 0.001) following treatment, associated with a significant fall (P < 0.05) in 24-h-long plasma glucose, FFA, and triglyceride concentrations. Neither circulating plasma insulin concentrations nor the ability of insulin to mediate glucose disposal changed with treatment. Finally, fasting total cholesterol fell (P < 0.05) and the ratio of total to HDL cholesterol decreased (P = 0.06) in association with Ergoset treatment. CONCLUSIONS: The fact that significant metabolic improvement was seen in the obese nondiabetic hyperinsulinemic women studied suggests that Ergoset could be of therapeutic benefit in clinical conditions of hyperglycemia and/or dyslipidemia. PMID- 9353613 TI - Reducing plantar pressure in the neuropathic foot. A comparison of footwear. AB - OBJECTIVE: To compare the effectiveness of therapeutic, comfort, and athletic shoes with and without viscoelastic insoles. RESEARCH DESIGN AND METHODS: We compared pressure reduction at ulcer sites under the hallux (n = 10), first metatarsal (n = 10), and lesser metatarsals (n = 12), using extra-depth, athletic, and comfort shoes with and without viscoelastic insoles. A rubber-soled canvas oxford was used to establish baseline pressure values. RESULTS: When used in conjunction with a viscoelastic insole, all shoe types reduced mean peak plantar pressure better than their non-insoled counterparts (P < 0.05). Consistently, comfort shoes reduced pressure significantly better than both the cross trainers and extra-depth shoes for ulcers under the first and lesser metatarsals (P < 0.05). For each shoe type, the addition of the viscoelastic insole provided a significant reduction in mean peak pressure (P < 0.05). Compared with stock insoles, viscoelastic insoles reduced pressures an additional 5.4-20.1% at ulcer sites. The same trend was also observed at regions of the foot not associated with an ulceration. CONCLUSIONS: When used in conjunction with a viscoelastic insole, both the comfort and athletic cross-trainer shoes studied were as, if not more, effective than commonly prescribed therapeutic shoes in reducing mean peak first and lesser metatarsal pressures. Furthermore, comfort shoes were as effective as therapeutic shoes in reducing pressure under the great toe. Both of these shoe types may be viable options to prevent the development or recurrence of foot ulcers. PMID- 9353614 TI - Impairment of peripheral blood flow responses in diabetes resembles an enhanced aging effect. AB - OBJECTIVE: To test the hypothesis that skin blood flow responses in the fingertip of diabetic patients are impaired and to examine the role of aging in both healthy control subjects and diabetic patients. RESEARCH DESIGN AND METHODS: We measured cutaneous blood flow using laser Doppler techniques in 40 people with diabetes and in 20 age- and sex-matched healthy control subjects. To induce vasoconstriction, subjects were asked to perform three 1-min stressor tasks: mental arithmetic, contralateral hand grip, and immersion of the contralateral hand in ice water. To induce vasodilatation, a local heat stimulus of 45 degrees C was applied for 5 min. RESULTS: Basal blood flow did not differ between groups, but vasoconstrictive responses induced by arithmetic or immersion of the contralateral hand in ice-cold water and vasodilatation induced by local heating were severely impaired in diabetic subjects, compared with healthy control subjects (P < 0.01). These responses correlated with autonomic nerve function and deteriorated significantly with advancing age in control subjects, but not in diabetic subjects. Blood flow in younger diabetic subjects resembled that of older control subjects. CONCLUSIONS: These data demonstrate that diabetes has effects on precapillaries that may by direct or mediated via autonomic nerves, which result in a deficit that resembles premature aging. PMID- 9353615 TI - Pronounced insulin resistance and inadequate beta-cell secretion characterize lean gestational diabetes during and after pregnancy. AB - OBJECTIVE: To evaluate beta-cell secretion and glucose metabolism in lean subjects with gestational diabetes mellitus (GDM) compared with that in subjects with normal pregnancy and obesity. RESEARCH DESIGN AND METHODS: Insulin secretion, insulin sensitivity (S1), and hepatic insulin extraction were assessed in pregnant women with GDM before and after delivery and in those with normal glucose tolerance (NGT) in comparison to healthy nonpregnant lean and obese women. Kinetic analysis of glucose, insulin, and C-peptide plasma concentrations during oral and intravenous glucose tolerance tests was performed by mathematical modeling. RESULTS: S1 was blunted in pregnant women with GDM by 84% and in those with NGT by 66% compared with lean nonpregnant women (P < 0.005 vs. healthy nonpregnant lean control subjects; P < 0.05, GDM vs. pregnant women with NGT), whereas glucose effectiveness was decreased by 33% in both pregnant groups (P < 0.05 vs. healthy nonpregnant lean control subjects). Insulin secretion was 30% higher (P < 0.05) in subjects with GDM than in pregnant women with NGT or in nonpregnant lean women, but decreased (P < 0.005) when compared with obese women with a comparable degree of insulin resistance. Fractional hepatic insulin extraction was similar in both pregnant groups, being lower (P < 0.0001) by 30% versus nonpregnant females. beta-cell sensitivity to glucose for insulin release was decreased in subjects with GDM versus pregnant women with NGT as well as nonpregnant women by 40-50% (P < 0.01). Twelve weeks after delivery, GDM returned to normal glucose tolerance, but S1 remained 50% lower than that in lean nonpregnant women, while beta-cell sensitivity to glucose did not change (P < 0.01 vs. healthy nonpregnant lean control subjects). CONCLUSIONS: Pregnancy is characterized by insulin resistance, diminished hepatic insulin extraction, and glucose effectiveness. Lean subjects with GDM are additionally characterized by having more pronounced insulin resistance and inadequate insulin secretion, which persist after delivery. Compared with other insulin-resistant prediabetic states like impaired glucose tolerance (IGT), defective insulin secretion seems to be a predominant defect in lean GDM subjects, indicating that it might represent a specific prediabetic condition. PMID- 9353616 TI - Visual function in young IDDM patients over 8 years of age. A 4-year longitudinal study. AB - OBJECTIVE: To carry out a longitudinal study of visual functions in young patients over the age of 8 years with IDDM and to assess the impact of metabolic control on the presence of diabetic retinopathy. RESEARCH DESIGN AND METHODS: There were 37 young IDDM patients from the Paediatric and Adolescent Clinic at the University Hospital of Wales studied annually for 4 years, with a control group of 24 healthy subjects observed over a 2-year period. Assessment of visual functions included visual acuity, color vision, and contrast sensitivity. Ophthalmoscopy and retinal photography were used to determine the presence or absence of diabetic retinopathy. In addition, pubertal status and metabolic control (glycosylated hemoglobin) were determined at each visit. RESULTS: Patients with IDDM demonstrate abnormal color vision and contrast sensitivity compared with the control group (P < 0.05), but visual acuity was unaffected. Visual functions were not significantly different between those IDDM patients with and without retinopathy. After 4 years, diabetic retinopathy was present in 43% of the group and was related to diabetes duration and metabolic control (P < 0.05). CONCLUSIONS: Visual function testing could not distinguish between those IDDM patients with and without retinopathy, but the color vision and contrast sensitivity in those with IDDM were significantly impaired compared with the control group. The presence of retinopathy was related to the duration of diabetes and metabolic control. It is important to ensure that good glycemic control and regular attendance for retinopathy screening is encouraged in the adolescent patients. PMID- 9353617 TI - Phenotypic expression of diabetes secondary to a T14709C mutation of mitochondrial DNA. Comparison with MIDD syndrome (A3243G mutation): a case report. AB - OBJECTIVE: To analyze the clinical and biochemical features of a recently described point mutation of mitochondrial DNA associated with diabetes. This mutation, characterized by a T14709C transition of a highly conserved nucleotide in the region coding for the glutamic acid tRNA, is heteroplasmic. RESEARCH DESIGN AND METHODS: The phenotypic expression in the insulin-requiring diabetic proband from the pedigree was compared to that of diabetic probands from three families with the classic A3243G mtDNA mutation (maternally inherited diabetes and deafness [MIDD] syndrome). The same investigations to evaluate pancreatic neurosensorial and muscle involvement were performed in all four patients. RESULTS: The natural courses of the diabetes and the hearing defects were not different between the two mutations. The patient with the 14,709 mutation, however, exhibited a milder alteration of pigmentary epithelium of retina and a much more severe muscle involvement, as attested by the clinical expression and the concurrent anomalies of muscle energy production evidenced by 31P magnetic resonance spectroscopy, confirming the profound impairment of oxidative processes. CONCLUSIONS: This novel mutation has to be added to the other known mtDNA anomalies in order to ascribe some diabetes suspected to arise from mitochondrial defects to this nosological framework. PMID- 9353618 TI - Insulin resistance and arteriosclerosis obliterans in patients with NIDDM. AB - OBJECTIVE: To investigate the risk factors for arteriosclerosis obliterans (ASO) in NIDDM, we measured insulin sensitivity and other risk factors including lipoprotein(a) [Lp(a)] in NIDDM patients with and without ASO. RESEARCH DESIGN AND METHODS: A case-control study in 100 patients with NIDDM, 35 with and 65 without ASO, was performed. Insulin sensitivity was assessed by the short insulin tolerance test's K index (KITT). Duration of diabetes, a history of smoking, prevalence of hypertension, prevalence of coronary artery disease (CAD), serum C peptide, 24-h urinary C-peptide, serum lipids, and Lp(a) were compared in the two groups. RESULTS: Age, BMI, HbA1c, and fasting plasma glucose were comparable in the two groups. Patients with ASO were significantly more insulin resistant than patients without ASO (KITT 2.16 +/- 0.16 vs. 3.00 +/- 0.13%/min, P < 0.0001, respectively), had a longer duration of diabetes (10.3 +/- 1.2 vs. 7.5 +/- 0.8 years, P < 0.05), included a greater number of smokers (68.6 vs. 40.0%, P < 0.01), had a higher prevalence of CAD (60.0 vs. 16.9%, P < 0.01), and had a greater percentage of insulin therapy (48.6 vs. 29.2%, P < 0.05). However, urinary and serum C-peptide levels, serum lipids, and Lp(a) levels were comparable in the two groups. Multiple logistic regression analysis indicated that a history of smoking (odds ratio 3.70, P = 0.011), insulin resistance (odds ratio 3.68, P < 0.001), and an elevated Lp(a) level (odds ratio 1.03, P = 0.020) were independently related to ASO. When patients with CAD were removed from the logistic regression analysis, insulin resistance was most strongly related to ASO (odds ratio 20.9, P < 0.001). CONCLUSIONS: Patients with ASO were characterized by a higher prevalence of CAD, a greater percentage of smokers, a greater percentage of insulin therapy, and a higher insulin resistance than were patients without ASO. Insulin resistance, especially, may be the most powerfully related to ASO. Lp(a) may play a minor role in the development of ASO. PMID- 9353619 TI - The prevention and treatment of obesity. Application to type 2 diabetes. PMID- 9353620 TI - Aspirin therapy in diabetes. PMID- 9353621 TI - Aspirin therapy in diabetes. American Diabetes Association. PMID- 9353622 TI - The role of viscous soluble fiber in the metabolic control of diabetes. A review with special emphasis on cereals rich in beta-glucan. AB - Recent recommendations for the dietary management of diabetes mellitus state that diet needs to be individualized so that there is improved glucose and lipid control in the patient. In a majority of individuals with diabetes, this is best done with a diet that is low in fat and high in carbohydrate, particularly that of cereal origin. However, symptoms of hyper- and hypoglycemia must be averted. Most cereal products, however, tend to have a high glycemic index Cereals such as Prowashonupana barley or fractions of oat bran are particularly high in the soluble fiber beta-glucan, which when taken with a meal increases the viscosity of the meal bolus once it has reached the small intestine, where the absorption of nutrients occurs. This high viscosity delays absorption. A 50% reduction in glycemic peak can be achieved with a concentration of 10% beta-glucan in a cereal food. A significant lowering of plasma LDL cholesterol concentrations can also be anticipated with the daily consumption of > or = 3 g of beta-glucan. Diabetic individuals can benefit from diets that are high in beta-glucan, which, as a component of oats and barley, can be incorporated into breakfast cereals and other products. PMID- 9353623 TI - Diabetes and accident insurance. A 3-year follow-up of 7,599 insured diabetic individuals. AB - OBJECTIVE: Individuals with diabetes pay increased premiums and experience limited coverage when taking out accident insurance, despite the lack of scientific support for this practice. The aim of the present study was to analyze whether diabetic individuals have an increased risk of accident or an increased risk of permanent disability after an accident, compared with two nondiabetic groups. RESEARCH DESIGN AND METHODS: All diabetic members of the Danish Diabetes Association were given free accident insurance for a 3-year period. Based on informed consent, they were also asked to participate in a follow-up study, comparing accident rates in diabetic individuals with a nondiabetic group. A total of 7,599 diabetic members accepted. The control groups were 1) individuals with a leisure-time insurance in the same company (individual issue) and 2) members of full-time group-based insurance (bank employees) in the same company. RESULTS: The risk of accidents was 0.7 per 1,000 person-years in the diabetic group, compared with 4.5 per 1,000 person-years in the first and 5.5 per 1,000 person-years in the second nondiabetic control group (P < 0.001). The degree of permanent injury did not differ between the diabetic and the nondiabetic group. CONCLUSIONS: The risk of accidents and permanent disability is not increased in diabetic individuals. Thus, diabetic individuals should be offered accident insurance at a standard premium without limited coverage. PMID- 9353624 TI - Diabetes in the Chinese population and its implications for health care. PMID- 9353625 TI - American Diabetes Association Annual Meeting, 1997. Type 2 diabetes. PMID- 9353626 TI - Mechanical misadministration of an oral hypoglycemic agent. PMID- 9353627 TI - Increased prevalence of NIDDM in anterior uveitis. PMID- 9353628 TI - The French paradox and diabetic patients. PMID- 9353629 TI - The tactile circumferential discriminator: an instrument for detecting patients at risk of foot ulceration. PMID- 9353630 TI - Incidence of type 1 diabetes in Germany is not higher than predicted. PMID- 9353631 TI - Mitochondrial DNA 3243 mutation is infrequent in Japanese diabetic patients with auditory disturbance. PMID- 9353632 TI - Intra-cage air change rate on forced-air-ventilated micro-isolation system- environment within cages: carbon dioxide and oxygen concentration. AB - Recently, a forced-air-ventilated micro-isolation system (FVMIS) has been recognized to accurately maintain microenvironmental conditions inside cages, but the details of the relationship between the concentrations of carbon dioxide (CO2) and oxygen (O2) and the air change rate inside the cages have never been reported. In this study, the proper intra-cage air change rate was examined based on the CO2 concentration and O2 concentration inside the cages measured by changing the ventilation volume inside the closed cages of the FVMIS while housing animals. In the experiments, three 8-week-old Wistar strain male rats weighing 303 g on average were housed in each FVMIS cage (capacity: 0.0223 m3), and the temperature, relative humidity, CO2 concentration and O2 concentration were measured when the air change rate inside the cages was varied from 10 air changes per hour (ACH) to 120 ACH. It proved that the CO2 concentration in the FVMIS cages decreased uniformly with the increase in the air change rate. As a result, 60 ACH was required to maintain the CO2 concentration level inside the FVMIS cages equivalent to or less than that in the conventional housing. Otherwise, when based on the O2 concentration, 50 ACH was required. In consideration of these results and others based on ventilation, airflow, temperature and the ammonia concentration reported previously, we concluded that the proper air change rate inside the FVMIS cages should be approximately 60 ACH. PMID- 9353633 TI - Immunohistochemical study on type II collagen-induced arthritis in DBA/1J mice. AB - We performed immunohistochemical examinations on type II collagen-induced arthritis (CIA) mice, focusing attention on the changes in distribution of plasma proteins and extracellular matrix materials (ECM) and in expression of adhesion molecules. The limb joints of male DBA/1J mice immunized with bovine type II collagen were obtained at 6 to 20 weeks after the first immunization. In the early stage of CIA, deposition of fibrin, IgG, von Willebrand factor (vWF) and fibronectin was detected on the surface of the synovial lining layer and articular cartilage and in the articular cavity. In the stage of pannus formation, prominent proliferation of ICAM-1-positive capillaries and marked infiltration of LFA-1-positive neutrophils were observed in the pannus. The superficial portion of the pannus and basement membranes of proliferated capillaries were strongly positive for type IV collagen and laminin. In the late stage, the pannus invaded and destroyed articular cartilage and subchondral bone, and strongly positive immunostainabilities for both lysozyme and fibronectin were observed on the surface of the pannus and at the junctional portion between the pannus and the cartilage. The present immunohistochemical findings on the distribution of plasma proteins and ECM materials and the expression of adhesion molecules in CIA mice were similar to those in rheumatoid arthritis (RA) in many aspects. This suggests that CIA is a useful model for the investigation of RA. PMID- 9353634 TI - Comparative morphometry of coxal joint angles. AB - The angles related to the coxal joints were comparatively studied in four-limbed walking animals and two-limbed ones including man and birds. Between animals with both types of walking, no significant difference was observed in the neck-shaft angles (NSA), which was equivalent to the acetabulum angles (ACA) at the connection of the femoral head with the acetabulum. The anteversion angles (AVA) were equivalent to the horizontal ACA. Canine species showed two different forms of the femoral neck with or without modification by the femoral AVA, probably being breed-specific and nutrition-dependent. In the narrow-striped wallaby as well as avian species, the femoral head showed a postversion with a minus-version angle for lifting the body axis in the frontal and upward direction to hold the whole body weight on the hind-limbs, in particular at the anterior part of the acetabulum. In man, the connection between the femur and acetabulum greatly varied among individuals, possibly according to differences in the life style. PMID- 9353635 TI - Response of respiratory epithelium of BN and F344 rats to formaldehyde inhalation. AB - BN rats are well-known for their high capacity for IgE production and hyperresponsiveness to exposure to allergens or other chemicals. We examined the histological changes in the nasal cavity, trachea and lungs of BN and F344 rats after the inhalation of aerosol formaldehyde (HCHO), which exerts direct toxic effects on the respiratory system. The incidence of clinical signs such as sneezing and abnormal respiration in HCHO-treated F344 rats was higher than that in HCHO-treated BN rats. The mean body weight of HCHO-treated F344 rats apparently decreased in comparison with control F344 rats, but that of HCHO treated BN rats was not significantly different from that of control BN rats. Changes such as squamous metaplasia, stratification, degeneration and desquamation were observed by light microscopy in nasal, tracheal and bronchial mucosa in the lungs of the HCHO-treated F344 rats. In the HCHO-treated BN rats, similar but milder lesions were restricted to the nasal mucosa. Scanning electron microscopic observation supported these light microscopic observations. These results suggest that BN rats have lower sensitivity to HCHO inhalation than F344 rats. PMID- 9353636 TI - Eurycoma longifolia Jack enhances libido in sexually experienced male rats. AB - The effects of Eurycoma longifolia Jack were studied on the libido of sexually experienced male rats after dosing them with 200, 400 and 800 mg/kg body weight twice daily of different fractions of E. longifolia Jack for 10 days. Results showed that E. longifolia Jack produced a dose-dependent increase in mounting frequency of the treated animals with 400 mg/kg of chloroform, methanol, water and butanol fractions resulting in mounting frequencies of 5.3 +/- 1.2, 4.9 +/- 0.7, 4.8 +/- 0.7 and 5.2 +/- 0.1, and 800 mg/kg further increased them to 5.4 +/- 0.8, 5.4 +/- 0.8, 5.2 +/- 0.6 and 5.3 +/- 0.2 respectively but there were no erections, intromissions, ejaculations or seminal emissions during the 20-min observation period which allowed for the measurement of sexual arousal reflected by mounting frequency uninfluenced by other behavioural components. This study provides evidence that E. longifolia Jack is a potent stimulator of sexual arousal in sexually vigorous male rats in the absence of feedback from genital sensation. PMID- 9353637 TI - The period of ovulation and presence of the first polar body of ova ovulated in the house musk shrew (Suncus murinus). AB - The period of ovulation in mature house musk shrews was examined in a natural mating group and a superovulation group treated with gonadotropin. In the natural mating group, ovulation started 14 hr after mating in 3 of the 7 house musk shrews (42.8%), and occurred in all 5 house musk shrews by 15 hr after mating. In the superovulation group, ovulation started 13 hr after the administration of hCG in 3 of the 5 house musk shrews, and was observed in all 5 shrews by 16 hr after the administration. In the natural mating group, ovulated ova were collected from the ovarian bursa of 14 house musk shrews 14-20 hr after mating (mean, 2.2 +/- 1.0 ova) and from the oviduct of 42 animals 14-24 hr after mating (mean, 3.6 +/- 1.8 ova). Among the ova ovulated 14-16 hr after mating, both mature ova with the first polar body and immature ova without the first polar body were observed. In the superovulation group, ovulated ova were collected from the ovarian bursa of 31 house musk shrews 13-22 hr after the administration of hCG (mean, 9.7 +/- 6.8 ova), and from the oviduct of 28 animals 13-24 hr after the administration of hCG (mean, 20.0 +/- 11.7 ova). There were also mature and immature ova in the ova ovulated 13-16 hr after the administration of hCG. The time when ova ceased to be recovered from the ovarian bursa roughly coincided with the time when new corpora lutea ceased to be found in the ovaries. These findings suggested that the period of ovulation of house musk shrews was 14-20 hr after mating in the natural mating group and 13-22 hr after the administration of hCG in the superovulation group. Both the natural mating group and superovulation group ovulated both mature ova with the first polar body and immature ova without the first polar body. PMID- 9353638 TI - Fiber digestion and weight gain in guinea pigs fed diets containing different fiber sources. AB - The effects of different fiber sources on feed intake, weight gain and digestibility of fiber were examined in guinea-pigs fed pelleted diets containing alfalfa meal, oaten hay, beet pulp and commercial hay cubes mixed with a basal diet at ratios of 3:1 (75% in the diet), 1:1 (50%) and 1:3 (25%). The basal diet contained 50.0% corn, 4.1% wheat, 22.1% wheat flour and 17.7% corn gluten meal. Food intake increased as the amount of fiber source was increased, but not in the case of beet pulp. The most digestible fiber (ADF and NDF) was that of beet pulp. Apparent digestibility of dry matter decreased with increasing ratios of fiber source to the basal diet for all fiber sources, but fiber and crude protein digestibilities varied and depended not only on the ratio of fiber to the basal diet but also on the source of the fiber. PMID- 9353639 TI - Subcapsular cell hyperplasia and mast cell infiltration in the adrenal cortex of mice: comparative study in 7 inbred strains. AB - Subcapsular cell hyperplasia (SCH) in the adrenal cortex of aged mice (13-15 months old) was frequent in both sexes of BALB/c, C3H/He, DBA/2J and IQI/Jic mice and in the females of A/J and C57BL/6, although the incidence and severity of SCH were considerably different among mouse strains. Mast cells were closely associated with SCH in the A/J, BALB/c, C57BL/6, DBA/2J and IQI/Jic mice, but not in the C3H/He strain. Compared with other strains, IQI/Jic mice had a significantly larger number of mast cells in the adrenal glands. Our findings suggest that mast cells may participate in the development of SCH, and IQI/Jic would be suitable for studying the pathogenesis of SCH and the role of mast cells in this lesion. PMID- 9353640 TI - Detection of nucleoprotein gene of Sendai virus in the lungs of rats by touchdown nested reverse transcription polymerase chain reaction. AB - The nucleoprotein (NP) gene of Sendai virus was detected by touchdown nested reverse transcription polymerase chain reaction (RT-PCR) in the lungs of a rat presented with respiratory illness and high serum ELISA titer to Sendai virus. This method seemed to be of value in controlling infection in laboratory rodents. PMID- 9353641 TI - Replication of murine coronaviruses in mouse embryonic stem cell lines in vitro. AB - Replication of murine coronaviruses in eight mouse embryonic stem (ES) cell lines of several genetic backgrounds was examined. Both mouse hepatitis virus (MHV) type 2 and MHV, strain A59 replicated well with no or minimal cytopathic effect in all the ES cell lines tested. The results suggest the possibility that MHV infected ES cells may disseminate MHV in mouse colonies due to embryo manipulation. PMID- 9353642 TI - Division of donor liver for successful split-liver transplantation in pigs. AB - Split Liver Transplantation (SLT) is an attractive method to solve the problem of a shortage of liver grafts. A through knowledge of the anatomy of the porcine liver vessels and bile duct is essential in performing the experimental SLT. This study was undertaken to decide the split line for successful SLT in pigs by examining the main branching patterns both vessels and bile duct in 30 porcine livers macroscopically and angiographically. The hepatic arterial branching patterns were divided into three types and bile duct patterns into two types. There was no exception in branching patterns of the portal vein and the hepatic vein. We conclude it is desirable that the donor liver should be divided into two grafts between the left medial lobe and quadrate lobe. PMID- 9353643 TI - Study on histamine related enzyme activities during murine hair cycle. AB - The beginning of each anagen phase of the hair growth cycle appears to partially repeat the stages in the initial development of the hair cycle, but the regulatory mechanism of the hair cycle is unclear. We have investigated the levels of histamine related enzyme activities in the third hair cycle period of C3H mouse after depilation. The level of histidine decarboxylase activity increased just after depilation treatment and returned to the normal level within two weeks: this change was relevant to histamine content as we have previously reported. This result suggests that the histamine synthesising enzyme, histidine decarboxylase, activity may be involved in the distinctive process of hair re growth, in particular, the initiation of the anagen phase. PMID- 9353644 TI - Pdx1, a homeodomain transcription factor required for pancreas development, maps to rat chromosome 12. PMID- 9353645 TI - What role for state health care in Asian transition economies? AB - The main findings of a study of the impact of changes to communism on health care in Asian transition economies are summmarized. PMID- 9353646 TI - Relative inefficiencies in production between solo and group practice physicians. AB - Health economists have hypothesized for some time that physicians produce medical care in an inefficient manner. Further, whether solo or group practice physicians are relatively more inefficient has been a question of particular interest. Theoretical considerations suggest that solo and group practice physicians face different behavioural and production constraints, implying that they may produce care at different levels of efficiency; which is more efficient is an empirical question. We employed stochastic production frontier estimation to address this issue. PMID- 9353647 TI - An incentive approach to physician implementation of medical practice guidelines. AB - We propose a probabilistically based incentive payment system for guideline implementation that provides rewards for physicians who follow practice guidelines and additional remuneration for physician leaders who engage in information sharing. All payments are based on observed outcomes of patient treatment. A fixed base payment forms the core of the system with probabilistic offsets calculated from the chance that a 'good' outcome occurs without optimal treatment or information. The system pays different physician types for different task sets. PMID- 9353648 TI - Lifetime costs of lung transplantation: estimation of incremental costs. AB - Despite an expanding number of centres which provide lung transplantation, information about the incremental costs of lung transplantation is scarce. From 1991 until 1995, in The Netherlands a technology assessment was performed which provided information about the incremental costs of lung transplantation. Costs in the situation with and without a transplantation programme were compared from a lifetime perspective. Because randomization was ethically inadmissible, only costs in the situation with the programme were observed. Both conventional treatment costs and costs of the transplantation programme were registered. Costs in the situation without the programme were based on the conventional treatment costs in the situation with the programme. Due to the study period of four years, long term follow-up costs were estimated. The total incremental costs per transplanted patient were estimated at Dfl 466,767 (5% discounted costs). The main part of these costs was caused by the high costs during the lifetime follow up of the patients. PMID- 9353649 TI - Trying to do better than average: a commentary on 'statistical inference for cost effectiveness ratios'. AB - In a recent paper, Laska, Meisner and Siegel address issues concerning hypothesis testing in cost-effectiveness analysis. They relate the relative magnitude of two average cost-effectiveness ratios to the incremental cost-effectiveness ratio and go on to propose a statistical procedure for testing the equality of two average ratios. In this paper, we show why the use of average cost-effectiveness ratios is misleading and argue that the appropriate focus for cost-effectiveness analysis is the estimation of confidence intervals around incremental cost effectiveness ratios. PMID- 9353650 TI - The usefulness of average cost-effective ratios. AB - We demonstrate that average cost-effectiveness ratios (CERs) play an important role in the evaluation of the cost-effectiveness of treatments. Criticisms of the usefulness of CERs derive mostly from the context of resource allocation under a constrained budget in which some decisions are based on incremental CERs. However, we show that in many cases, these decision rules are equivalent to decision rules on CERs. This follows for mutually exclusive treatments first, because a treatment is eliminated by extended dominance if and only if there is a mixed treatment with a smaller CER, where the mixing parameter lies in a certain interval. Second, after elimination of treatments by dominance and by extended dominance, resources can be allocated in order of increasing CERs. Moreover, the CER is a parameter that characterizes clinical and economical properties of a treatment independent of its comparators. PMID- 9353652 TI - Productivity costs in cost-effectiveness analysis: numerator or denominator: a further discussion. AB - In this response we concentrate on what Weinstein et al. call the 'major disagreement' between the Erasmus group and the US Panel, which concerns the measurement of productivity losses during illness. We consider the consequences for the individual, for the employer and for the rest of society and argue that when following the Panel's propositions for measuring these consequences, major theoretical and practical difficulties are encountered. PMID- 9353651 TI - Productivity costs, time costs and health-related quality of life: a response to the Erasmus Group. PMID- 9353653 TI - Values and preferences are not necessarily the same. AB - Economic theory typically draws no distinction between preferences and values, assumes that preferences are stable and complete and that all that need be done to elicit them is to ask the right question in the right way. It is argued here that values for some fundamental aspects of life, such as health, are not the same as preferences. The former are less differentiated and require construction and clarification before they can be elicited. The implications of this for health state valuation are discussed. PMID- 9353655 TI - Economic modelling of the gateway effect. AB - Although a significant number of empirical studies provide evidence of sequencing in drug use, economic theory remains focused on addiction to a single substance. This paper presents a general model of substance use that allows for the possibility of multi-commodity habit formation and can be used to analyse the intertemporal relationship between the consumption of legal and illicit drugs, or the gateway effect. A simple two-drug model is analysed and conditions for the existence of multi-commodity habit formation are examined. It is found in the case of multi-commodity habit formation that the marginal utility of initiating a new drug is higher when there is prior consumption of the other drug. Further, it is found that the individual will initiate drug consumption with that drug that has the lowest marginal cost. The particular sequencing of drug use that is observed in empirical data is explained by differences in the marginal cost of consuming legal and illegal drugs. PMID- 9353656 TI - Effects of tobacco excise taxes on the use of smokeless tobacco products in the USA. AB - Data from the September 1985 Current Population Survey are used to estimate the effects of tobacco excise taxes and state laws restricting smoking in public places on the likelihood of current use of cigarettes or smokeless tobacco (ST) products (moist snuff or chewing tobacco) among males in the USA. The results indicate that higher ST excise tax rates are associated with a reduced probability of ST use, whereas higher cigarette excise tax rates are associated with an increased probability of ST use, holding other factors constant. State laws restricting smoking have no apparent effect on ST use. PMID- 9353657 TI - Women and work: tipplers and teetotalers. AB - We seek to understand better the puzzling finding that, for women, alcoholism appears to be positively associated with the probability of being employed. Using the 1988 Alcohol Survey of the National Health Interview Survey, we find that this association holds for white women only. For white women, alcoholism and early drinking are associated with higher educational attainment, a smaller family size and a lower probability of being married. In turn, these human capital indicators are associated with greater labour supply, thus helping to explain the curious positive relationship between alcoholism and employment for women. An advance in this paper over our previous work is to examine life-time abstention from alcohol and its association with employment and human capital variables. We find that lifetime abstention is associated with lower employment, unemployment and education and greater propensity to be married for both white and non-white women. PMID- 9353658 TI - Estimating the economic cost of substance abuse treatment. AB - Few studies have estimated the economic costs and benefits of substance abuse treatment services. This paper introduces a data collection instrument and method for estimating the economic cost of substance abuse treatment programs. The Drug Abuse Treatment Cost Analysis Program (DATCAP) is based on standard economic principles and the method has recently been tested in two drug abuse intervention studies. Findings from case studies at three treatment programmes are presented to demonstrate the feasibility and reliability of the instrument. The estimation methods and results can be used by treatment programmes for self-evaluation purposes and by researchers who are interested in performing cost-effectiveness or benefit-cost analyses of substance abuse services. PMID- 9353659 TI - Social inequalities and cancer. PMID- 9353660 TI - Why study socioeconomic factors and cancer? AB - The occurrence of cancer within a population can be studied at many different levels, including forms of social organization, the individual, a particular organ system, or a particular molecule. The causes of cancer can also be studied at these different levels, including socioeconomic factors, lifestyle, the organ burden of a carcinogen, or DNA adducts. Clearly, there are advantages in understanding disease causation at all of the different levels at which it operates. Although cancer risk factors such as tobacco smoke may appear to operate at the individual level, exposure may occur due to a wide range of political, economic and social factors; conversely, tobacco smoke ultimately also has effects at the cellular and molecular levels, including the production of mutations in DNA. Of course, it is important to gain information, and take action, at all possible levels, but the history of public health shows that changes at the population level are usually more fundamental and effective than changes at the individual level, even when a single risk factor accounts for most cases of disease. In this sense, a risk factor such as smoking can be regarded as a secondary symptom of deeper underlying features of the social and economic structure of society. Thus, just as a variety of health effects in various organ systems (for example, various types of cancer) may have a common contributing cause (for example, tobacco smoking) at the level of the individual, a variety of individual exposures (for example, smoking and diet) may have common socioeconomic causes at the population level. In many instances there is clear evidence that cancer is related to socioeconomic factors, but this does not appear to be fully explained by known risk factors. More importantly, there is little evidence as to which socioeconomic factors are of most importance, or whether it is the overall 'package' of social inequality that is responsible for the differences in cancer risk. The aim of this book is therefore to summarize what is already known, and to identify gaps in our knowledge. PMID- 9353661 TI - Poverty and cancer. AB - Despite the attraction of certain utopias and the convincing strength of some of the social and philosophical theories underlying attempts to change the social structure and to achieve a more egalitarian society, social inequalities have not disappeared and seem even to be increasing worldwide. Inequalities in health are part of the social inequalities present in our society and one of their most convincing indices. Sanitary conditions are worse, mortality higher, survival rates of cancer patients lower, and life expectancy shorter in developing countries than in industrialized countries. Similar if not identical differences can be seen within industrialized countries between socioeconomically less and more favoured population groups. In many areas of the industrialized countries social and environmental conditions comparable with those existing in the poorest countries last century have been recreated. Occupational risks are becoming a serious problem in developing countries, largely as a consequence of the transfer of hazardous industries from industrialized countries where certain industries are judged to be unacceptable. A similar double standard is applied to tobacco advertising and sales in the industrialized and developing countries. The projections of the total number of cancer cases in the next decades indicate a generalized increase, proportionally greater in developing than in industrialized countries. PMID- 9353662 TI - Social theory and social class. AB - Concepts of class developed with the emergence of industrial society in the nineteenth century. For an understanding of current divisions, theories must reflect the advances of capitalism and the global economy that characterize the late twentieth century. In industrialized societies, reductions in the industrial workforce and the growth of finance, investment and real-estate industries worldwide have produced a new, largely female, service workforce. Large sectors of industry have departed in search of cheaper labour in poorer countries, which also have a rising number of women workers. In those areas, as a result, a new industrial workforce has emerged. Concomitantly, accumulation of land in less developed agricultural regions for production for the world market has led to an increase in mobile agricultural labour and a shift of landless labourers to the cities of less developed countries. In addition, both upward and downward mobility have occurred for individuals and groups in specific populations, as well as for particular diseases in developed and less developed countries. All these processes have precipitated fundamental changes in class, gender and family relationships and transformed the living conditions of populations in both developed and less developed societies. These changes have major implications for the patterns of health and disease in the world today. Objective measures of social change may be difficult to construct and use in epidemiological cancer research. Since questions of class and shifting social relations are directly implicated in the patterns of disease, they must be assessed in future research as accurately as possible. PMID- 9353663 TI - The measurement of social class in health studies: old measures and new formulations. AB - The measurement of socioeconomic status (SES) is a serious matter that requires us to think more precisely about both conceptual issues and issues more traditionally thought of as measurement issues. Progress in this area rests on our ability to identify those aspects of SES that are most closely related to health, human development, and life expectancy. In this chapter we review measures of SES based on characteristics of the individual as well as on characteristics of the environment or more ecologically based measures. Each of these types of SES measures has strengths and weaknesses and in all likelihood taps somewhat different aspects of class. In measuring SES across diverse populations, it is also crucial to be sensitive to the ways in which measurement varies across different cultural, ethnic and demographic groups. It is likely that as we conduct more refined research in this area we will understand more fully why SES is so profoundly related to health status. However, so as to understand this relationship, we will need to expand efforts to identify not only those psychosocial or biological processes that occur 'downstream' as a result of SES but also the nature of the social experience itself and those 'upstream' forces that place so many individuals at risk. PMID- 9353664 TI - Socioeconomic differences in cancer incidence and mortality. AB - This chapter summarizes accumulated data on the presence, magnitude and consistency of socioeconomic differentials in mortality and incidence of all malignant neoplasms and 24 individual types of neoplasms in 37 populations in 21 countries. More or less consistent excess risks in men in lower social strata were observed for all respiratory cancers (nose, larynx and lung) and cancers of the oral cavity and pharynx, oesophagus, stomach, and, with a number of exceptions, liver, as well as for all malignancies taken together. For women, low class excesses were consistently encountered for cancers of the oesophagus, stomach, cervix uteri and, less consistently, liver. Men in higher social strata displayed excesses of colon and brain cancers and skin melanoma. In the two Latin American populations for which data were available, lung cancer was more frequent in higher social strata. Excesses in high female socioeconomic strata were seen in most populations for cancers of the colon, breast and ovary and for skin melanoma. Longitudinal data from England and Wales suggested widening over time of social class differences in men for all cancers combined and for cancers of the lung, larynx and stomach, and in women for all cancers combined and for cervical cancer. PMID- 9353665 TI - Socioeconomic differences in cancer survival: a review of the evidence. AB - In the discussion of social inequalities in health there has been much debate on the role of medical care. Large differences in cancer incidence and mortality from cancer have been consistently observed. To understand the potential importance of socioeconomic differences in prompt detection and treatment of cancer it is essential to have data on cancer survival. These have been examined less extensively than differences in cancer incidence. We have reviewed 42 studies on social class differences in cancer survival. Twenty-three studies were conducted in North America, and 15 in western European countries. Twenty-three studies were carried out through population-based cancer registries and 17 through hospitals or hospital-based registries. Seven studies examined survival differences for multiple cancer sites. Social class differences in cancer survival appear remarkably general. Patients in low social classes had consistently poorer survival than those in high social classes. The magnitude of the differences for most cancer sites was fairly narrow, with most relative risks falling between 1 and 1.5. The widest differences were observed for cancers of good prognosis and specifically cancers of the female breast, corpus uteri, bladder and colon. The pattern of the social differences in survival did not vary consistently by sex, country, or source of the study population and did not depend on the socioeconomic indicator used. PMID- 9353666 TI - General explanations for social inequalities in health. AB - Life expectancy has always differed according to status in society, with a higher mortality among those of lower social status. Although cancer and cardiovascular diseases are more common as causes of death in rich than in poor societies, in industrialized countries the major causes of death are more common in those of lower social status. In this chapter, the magnitude of socioeconomic differences in health is examined using different measures of socioeconomic status, and methodological issues relating to these measures are discussed. Much of the discussion about social inequalities in health has been focused on the health disadvantage of those of lowest socioeconomic status. However, data from the Whitehall studies show that the social gradient in morbidity and mortality exists across employment grades in British civil servants, none of whom is poor by comparison with people in developing countries, suggesting that there are factors that operate across the whole of society. A number of potential explanations are considered here. The magnitude of socioeconomic differences in health varies between societies, and over time within societies. This suggests that identification of factors that influence socioeconomic status and health, and the pathways by which they operate, is an important public health task that could lay the basis for a reduction in inequalities in health. PMID- 9353667 TI - Tobacco smoking, cancer and social class. AB - Consumption of tobacco products, both by smoking and by other means, has long been causally connected with cancers of the lung, larynx, mouth and pharynx, oesophagus, bladder, and many other sites. Tobacco is the main specific contributor to total mortality in many developed countries and has become a major contributor in the developing countries as well. In most industrialized countries, prevalence of cigarette smoking is currently higher in low than in high social classes, although in some industrialized countries smoking was more frequent in high social classes during the first half of this century. The latter pattern of tobacco consumption is more likely to apply to developing countries. To formulate and carry out effective tobacco control activities it is essential to assess the relative incidence of tobacco-related cancers in different social strata and the prevalence of tobacco use across strata. Despite many years of data gathering the information base is far from complete, especially in developing countries where tobacco use is increasing rapidly, and where aggressive marketing by the transnational tobacco industry is occurring. A critical question is the extent to which tobacco usage can 'explain' the observed social class differences in cancer risk. Class differences in lung cancer are likely to be mostly related to the unequal distribution of tobacco smoking between social classes, and in some fairly simple situations this has been satisfactorily demonstrated. Nevertheless, there are many unresolved issues, especially with regard to the role of collateral exposures, such as hazardous occupations, poor diet, and limited access to health care. The question of whether tobacco use 'explains' socioeconomic differences in one or more of the cancers that it causes has rarely been directly addressed in epidemiological studies. PMID- 9353668 TI - Alcohol drinking, social class and cancer. AB - This chapter reviews the data on occurrence of cancers that are potentially caused by alcohol drinking (cancers of the upper gastrointestinal and respiratory tracts, and liver cancer) in relation to social class. In order to assess the role of alcohol drinking in the observed social class gradients of these cancers, we have particularly looked for consistency in the gradients of different alcohol related cancers, and used lung cancer occurrence to judge the role of tobacco smoking, which is the major other determinant of these diseases. Additional data on levels of alcohol drinking and on the occurrence of other alcohol-related morbidity are brought into the discussion where available. A role of alcohol drinking in the observed negative social class gradients for alcohol-related cancers is very likely in men in France, Italy and New Zealand. Evidence that is less strong, but is suggestive of a role of alcohol drinking, is seen for men in Brazil, Switzerland, the United Kingdom and Denmark. Although a role of alcohol drinking is likely or possible in certain populations, other factors may contribute as well, most notably tobacco smoking and dietary habits. Additional data on the frequency of complications after surgical procedures in alcohol drinkers are reviewed briefly. PMID- 9353669 TI - Diet and cancer: possible explanations for the higher risk of cancer in the poor. AB - Humans have always had to eat; diets have always contained the same nutrients and bioactive constituents. Therefore, some have argued, the present pattern of diseases and changes in that pattern cannot be causally linked to dietary intake. This argument, its naivety notwithstanding, raises some important issues for the way we think about the epidemiology of nutrition and disease. Current research on diet and specific diseases is based, obviously, on the premise that this argument is false. This chapter uses a broad brush to present the evidence for a significant and causal association between eating patterns and cancer. It shows that, far from being an implausible link, the relationship between dietary patterns and cancer is largely explained by the dependence of humans on their food supply--dependence not merely in the sense of providing energy to sustain life, but more related to evolutionarily adaptive patterns of food intake and contemporary aberrations in those patterns. The chapter also shows that it is plausible that at least part of the explanation for the higher risk of cancer among the poor in both rich countries and poor countries relates to the extent of the aberrations in food supply and eating patterns. PMID- 9353670 TI - Socioeconomic differences in reproductive behaviour. AB - There are marked socioeconomic variations in the risk of female reproductive cancers. We examine here data from the World Fertility Surveys, the Demographic and Health Surveys, and other national surveys, to assess whether these variations in cancer risk might be explained, at least in part, by socioeconomic variations in reproductive behaviour. There were marked socioeconomic differentials in achieved parity, age at first birth, final childlessness, duration of breastfeeding, and possibly also age at menopause. These differentials were present in almost all settings: countries with low and high levels of modernization, and countries with low and high levels of fertility. In general, women of higher socioeconomic status and with more education had lower fertility and later age at first birth, but a greater prevalence of childlessness, shorter duration of breastfeeding and later age at menopause. However, the size and even the direction of these differentials varied markedly from country to country according to its level of economic development and, within each country, from generation to generation of women. It is possible that some of these socioeconomic differences may be narrowing in recent generations in Western countries. There was little evidence of socioeconomic variations in age at menarche. The observed socioeconomic differentials in most aspects of reproductive behaviour could potentially account for some of the socioeconomic variation in the risk of female reproductive cancers. However, this relationship could not be assessed directly because such analysis would require birth-cohort specific data on socioeconomic variations in reproductive behaviour and in cancer risks. Unfortunately, these data are not available. PMID- 9353671 TI - Social differences in sexual behaviour and cervical cancer. AB - In this chapter we first describe the variation of cervical cancer in relation to social class. Thereafter we examine the causes for the occurrence of socioeconomic differences in invasive cervical cancer, using data from two case control studies carried out in Colombia and Spain. Cervical cancer is the most common cancer in developing countries and the sixth most common in developed countries. In all areas, it is more frequent among women of low socioeconomic status, it is associated with multiple sexual partners and early age at first sexual intercourse, and both incidence and mortality are reduced by screening. According to population-based surveys in industrialized countries, men of low socioeconomic status report fewer sexual partners than men of high socioeconomic status but there is no clear indication that the same is true of women of low socioeconomic status. In the case-control studies in Spain and Colombia, the human papillomavirus and all other sexually transmitted diseases were more prevalent among women in low socioeconomic strata. Number of sexual partners and particularly contacts with prostitutes were higher among husbands of women of low socioeconomic status. Other potential risk factors for the disease, such as smoking and oral contraceptive use, and also cervical cancer screening (Pap smears), were more common in women of high social strata. Women with no schooling had a threefold higher risk in Spain and a fivefold higher risk in Colombia of having cervical cancer compared with women who had achieved a higher educational level. After adjustment for sexual behaviour, HPV DNA status, history of Pap smears and husband's contact with prostitutes, this association was considerably reduced. These results are indicative that socioeconomic differences in the incidence of cervical cancer can be partly explained by differences in the prevalence of HPV DNA. Men's sexual behaviour and particularly contacts with prostitutes might be a major contributor to the higher prevalence of HPV DNA among the poor. PMID- 9353672 TI - Infection with hepatitis B and C viruses, social class and cancer. AB - The hepatitis B and C viruses (HBV and HCV) are major etiological factors in the occurrence of hepatocellular carcinoma (HCC) worldwide, but most especially in developing countries where the majority of liver cancer cases can be found. In parallel with the geographic distribution of HCC, high levels of HBV endemicity are concentrated in the developing world. The association between chronic infection with HBV and low social class is quite strong; socioeconomic factors such as low educational attainment, lower social stratum, and crowded urban residence have been reported to predict higher HBV chronic carrier prevalence in both developed and developing countries. More importantly, the effect of poverty on HBV endemicity is clearly evident among younger age groups, and earlier chronic HBV infection seems to increase the risk of development of HCC. As assays for detecting HCV antibodies have only recently become available, the data on the relationship between HCV infection and socioeconomic status are much fewer. However, the limited number of studies that have investigated the seroepidemiology of HCV report an association between higher prevalence of antibodies to HCV and indicators of low social class. It would appear that the striking correlation between HCC and low socioeconomic status is largely related to the impact of poverty on the spread of HBV and probably HCV. PMID- 9353673 TI - Infection with Helicobacter pylori and parasites, social class and cancer. AB - Three genera of parasites are known or suspected risk factors for cancer in humans: Schistosoma, Opisthorchis and Clonorchis. No adequate information is available on the determinants of infections related to social class. Infection with the bacterium Helicobacter pylori is an important cause of stomach cancer. Studies, in particular from the United Kingdom and the United States of America, strongly suggest that social class factors, especially those acting during childhood, are determinants of the infection, with odds ratios of seroprevalence of the order of 1.5-5 for lower social class as compared with higher social class. A conservative estimate of the contribution of social class, acting through an increased prevalence of H. pylori infection, to the burden of stomach cancer gives a figure of over 50,000 stomach cancers per year worldwide, or 8% of all stomach cancers. In countries with both high and low prevalence of infection with H. pylori, it is likely that a sizeable proportion of this difference is due to social-class-related risk factors of infection. PMID- 9353674 TI - Exposure to occupational carcinogens and social class differences in cancer occurrence. AB - It has been estimated that occupational exposures are responsible for about 4% of all human cancers in industrialized countries. These cancers are concentrated among manual workers and in the lower social classes, thus contributing to the social class gradient in cancer incidence and mortality. On the basis of the 1971 cancer mortality data from England and Wales, it was estimated that occupational cancer is responsible for about a third of the total cancer difference between high (I, II and III-NM) and low (III-M, IV and V) social classes, and for about half of the difference for lung and bladder cancer. However, direct evidence on the extent of the contribution of occupational exposure to carcinogens to social class differences is lacking, and several problems, such as the possible interaction between carcinogens and the effect of extraoccupational confounding factors, add further elements of uncertainty. PMID- 9353675 TI - Unemployment and cancer: a literature review. AB - With a tenth of the labour force involuntarily out of work, unemployment has become an important element among the socioeconomic determinants of health in the rich countries. Unemployed men have an excess cancer mortality of close to 25% compared with that of all men in the labour force. The available data from various countries indicate that this excess risk is found both in periods when the unemployment rate is about 1% and in periods when it is about 10%. Furthermore, it persists long after the start of unemployment and it does not disappear when social class, smoking, alcohol intake, and previous sick days are controlled for. The excess cancer mortality comes mainly from lung cancer, and the increased risk of lung cancer does not disappear when social class and number of previous sick days are controlled for. Unemployment does not increase smoking, but unemployed men have a slightly higher smoking prevalence before unemployment. However, as the excess lung cancer risk among unemployed men remains after controlling for social class, it seems unlikely that it can be explained only by differences in smoking prior to unemployment. PMID- 9353676 TI - Unemployment and cancer in Denmark, 1970-1975 and 1986-1990. AB - We have analysed cancer mortality and cancer incidence among unemployed persons identified from the Danish linkage studies based on the 1970 census and the 1986 register-based census. In 1970, 1% of Danish men were unemployed; in 1986, 14% were unemployed. In both periods, unemployed men had an excess cancer mortality of close to 25% when they were followed-up for a five-year period and their mortality was compared with that of all men in the labour force. Unemployed women in the 1970 cohort also had an excess cancer mortality of 25%. Cancer incidence data were not available for the 1986 cohort. For both cohorts, the excess risk came mainly from lung cancer. Survey data from Denmark in the 1980s indicated that unemployed men had a slightly higher smoking prevalence before unemployment than men who continued working, and that unemployment did not increase smoking. It is therefore unlikely that the excess lung cancer risk among unemployed men is explained by differences in smoking habits alone. PMID- 9353677 TI - Environmental exposure, social class, and cancer risk. AB - Exposure to a variety of environmental factors associated with cancer occurrence varies by social class. These factors include air pollutants (SO2, NO2, total suspended particulates, etc.), toxic waste hazards, and ionizing and other radiation. Heavy environmental pollution has been associated with an increased risk of some cancers and in particular lung cancer. There is limited evidence suggesting that individuals from lower social classes are exposed to higher levels of environmental pollutants than are individuals from higher social classes. This may be due to the placement of new sources of pollution or of toxic processes in disadvantaged areas, or to the selective migration of the poorer sectors of society to these areas. The available data do not allow any conclusion on the possible contribution of exposure to environmental pollution to social class differences in cancer occurrence. Exposure to ultraviolet (UV) radiation, principally from sunlight, is modified strongly by personal behaviours such as choice of recreation and use of protective clothing. Those in outdoor occupations are likely to receive the highest cumulative exposure to UV radiation. There is no clear evidence from recent survey research in Australia and North America that socioeconomic factors are strongly related to non-occupational exposure to UV radiation. Information is lacking on the influence of socioeconomic status on sun exposure in other parts of the world. There is little information on the social distribution of exposure to ionizing radiation. PMID- 9353678 TI - Socioeconomic status and cancer screening. AB - The only widely applied cancer screening programmes are those for cancers of the cervix and female breast. Participation in breast cancer screening has been shown to depend on income and education, health insurance and type of health service. Women in low social classes tend to have lower screening participation rates than those in higher classes. Socioeconomic differences in screening practices tend to decrease when participation is promoted, cultural and economic barriers are removed, and social support is offered. In both developed and developing countries, women of low socioeconomic status have a higher than average risk of cervical cancer, and a lower than average participation in Pap smear screening. PMID- 9353679 TI - Possible explanations for social class differences in cancer patient survival. AB - Social class differences in cancer patient survival have been reported for most cancer types and for a number of countries. The etiology of these differences has been studied less thoroughly and less systematically than social class differences in cancer occurrence. Stage of disease at diagnosis appears to be the most important factor contributing to the social class differences in cancer patient survival. This has been observed most clearly for gastrointestinal and gynaecological cancers. Social class differences in survival are generally wider for patients diagnosed with cancer at local stages than for those diagnosed with cancer at advanced stages. The reasons why cancers are more frequently diagnosed at a local stage in high than in low social classes in not properly understood at the moment. Of other potential contributing factors, the role of treatment and psychosocial factors has scarcely been studied. Biological indicators of tumour aggressiveness have failed to explain the social class differences. PMID- 9353680 TI - Family practice physicians' firearm safety counseling beliefs and behaviors. AB - The purpose of this study was to identify family physicians' firearm safety counseling beliefs and behaviors. A survey was mailed to a random sample of 600 members of the American Academy of Family Physicians. A three wave mailing technique was used to maximize the response rate and yielded 271 usable surveys (55% response rate). Outcome measures included training experience in firearm safety counseling, the prevalence of firearm safety counseling by family physicians, and their perceptions regarding such counseling. The majority (78%) of family physicians lacked formal training on how to counsel patients about firearm safety and 49% believed more time should be spent in residency programs on firearm safety counseling. The majority (84%) of respondents never or rarely counseled patients on firearm safety and 50% believed firearm safety counseling should be a low priority in their delivery of primary care. The majority of respondents did not regularly counsel patients about firearm safety, did not believe firearm safety counseling should be a priority, and did not believe firearm safety counseling would be effective in reducing firearm-related trauma. PMID- 9353681 TI - Results of a community-based low-literacy nutrition education program. AB - A nutrition intervention focused on low-fat eating pattern changes was conducted among low-literacy participants in a Twin Cities Metropolitan area Expanded Food and Nutrition Education Program (EFNEP). A total of 134 EFNEP participants who participated in the intervention were compared to 70 comparison participants who received EFNEP nutrition education materials. Associations between changes in outcome variables specific to the intervention were evaluated using mixed-model regression analyses. The principal effects seen for this program were related to changes in eating pattern scales. More modest effects were seen in scales related to attitudes of low-fat eating, and although changes in dietary fat intake as measured by 24-hour dietary interviews suggested a positive intervention effect, this did not approach statistical significance. PMID- 9353682 TI - Evaluating a statewide partnership for reducing risks for chronic diseases. AB - We describe a case study evaluation of Kansas LEAN, a statewide partnership with the mission of reducing risks for chronic diseases through dietary and exercise modification. We used a case study design to examine five primary questions related to process and outcome: (a) were the goals of the partnership important to constituents? (process), (b) were constituents satisfied with the partnership (process), (c) were community or systems changes (new or modified programs, policies, or practices) facilitated by partnership efforts (outcome)?, (d) were these changes important to the partnership's mission (outcome)?, and (e) what critical events helped facilitate community changes (outcome)? several measurement instruments--a monitoring and feedback system, constituent surveys, and semistructured interviews--were used to address key evaluation questions. Kansas LEAN is a strong statewide partnership with involvement from key representatives throughout Kansas. It is an ongoing, comprehensive health promotion program that plans and implements multiple components, in a variety of settings, to create awareness, behavior change, and a supportive environment. Kansas LEAN has facilitated several important community or systems changes related to its mission. We conclude with a discussion of the challenges of evaluating partnerships that seek to reduce risks for chronic diseases. PMID- 9353683 TI - Hispanics and worksite health promotion: review of the past, demands for the future. AB - During the last 15 years, reports, books, and published papers have demonstrated the positive health benefits of comprehensive health promotion and disease prevention interventions at the worksite. Although the progress made in worksite health promotion should not be understated, experts agree that it is time to step forward and address the new demands of the changing labor force. One area of intervention that needs to be more aggressively addressed is that of minority populations. Because work force projections show the high participation rate that Hispanics will have in the future labor force, the Hispanic population should be one of the targets of worksite health promotion. This paper reviews how employee health promotion programs have addressed Hispanic workers in the past and establishes directions for the future. PMID- 9353684 TI - Skin cancer prevention education: a national survey of YMCAs. AB - High skin cancer incidence and mortality rates have created a need for skin cancer prevention education. Children are an important target for this education, as overexposure to sun and sunburns at an early age have been linked to the development of skin cancer. This study identified the prevalence of skin cancer prevention education and the need for this education at YMCA swim classes. This study also assessed the feasibility of implementing Project SUNWISE, an existing skin cancer prevention curriculum, developed for YMCA swim classes. A 51-item survey was mailed to Aquatics Directors at all YMCAs with outdoor pools. Based on a 63% response rate (N = 208), only 28% of YMCAs thought the children in their swim classes were adequately protected from the sun, and only 28% offered skin protection or sun safety education in swim classes. Only 50% of YMCAs trained their swim instructors on skin cancer prevention. While 91% of YMCAs had one or more sun protection items near the pool (e.g., sunscreen, umbrella, covered area), 93% of YMCAs saw the need for additional protection. The majority of YMCAs (95%) were willing to incorporate a skin cancer prevention education curriculum, similar to Project SUNWISE, into their swim classes. This study emphasized the need for more skin cancer prevention education programs targeting children and examined the correlates of skin cancer prevention education at YMCAs. Geographic region, percent of possible sunshine, and ultraviolet radiation were significantly associated with the skin cancer prevention education program status at the YMCAs. PMID- 9353685 TI - Health care coverage: traditional and preventive measures and associations with chronic disease risk factors. AB - Physician counseling of patients on health related activities is an essential component of chronic disease prevention, however this requires patients to have ready access to health care providers. Previous studies have explored access to health care in terms of health plans and cost without accounting for the lack of preventive coverage inherent in many insurance policies. This study compares two measures of health care access, one using an assessment of cost and health plan availability, and a new coverage measure including preventive services. Data was collected from 2574 adult respondents to the 1991-92 Missouri Behavioral Risk Factor Surveillance System Surveys. Odds ratios were generated for demographic variables, health related behaviors and preventive screening and the two coverage measures. Using health plan and cost 22% lacked full coverage, however including availability of preventive coverage almost 60% lacked full coverage for preventive care. For both coverage measures significant associations were found with age, exercise, marital status, routine checkup and mammography screening. Using the measure of coverage of preventive services, rural residents and those who had never had cholesterol screening were more likely to lack coverage. Inclusion of preventive care in measures of health care coverage may alter previously reported associations with socio-demographic and health related factors. Policy makers should realize that including preventive services in health care coverage greatly increases the number of individuals lacking adequate coverage, and that those lacking adequate coverage are the least likely to undergo preventive screening. PMID- 9353686 TI - Thigh muscle size and strength after anterior cruciate ligament reconstruction and rehabilitation. AB - It is the hypothesis of the senior author (GAA) that high circumference measurements are not an accurate reflection of thigh muscle cross-sectional area or muscle strength after standard rehabilitation following anterior cruciate ligament reconstruction. Likewise, normal quadriceps femoris strength is not achieved in these patients despite aggressive rehabilitation. The purpose of our study was to quantify thigh muscle size and strength and correlate thigh circumference, muscle cross-sectional area by magnetic resonance imaging (MRI), and isokinetic strength in our patients. Thirty-three patients with anterior cruciate ligament repair utilizing autografts of iliotibial band (N = 28), semitendinosus autograft (N = 3), and bone-patellar tendon-bone autograft (N = 2) were retrospectively evaluated 48.7 +/- 6.91 months after surgery. We compared involved operated extremities with uninjured, uninvolved contralateral extremities, measuring thigh circumference, isokinetic peak torque, and cross sectional area by MRI. We found a significant 1.8% decrease in thigh circumference, a 10% decrease in average quadriceps torque, and a 8.6% decrease in quadriceps cross-sectional area by MRI in the involved extremities compared with the uninvolved extremities. A positive correlation between MRI cross sectional area, quadriceps, and hamstring peak torque was recorded in involved and uninvolved extremities. A positive correlation between thigh circumference, quadriceps, and hamstring peak torque was found in uninvolved extremities but not in operated extremities. The authors concluded that thigh circumference underestimates atrophy and is not correlated with cross-sectional thigh muscle area by MRI or strength in operated extremities. Persistent quadriceps weakness and decreased cross-sectional area at 49 months postsurgery and rehabilitation continue to challenge our efforts. The pathophysiology of the decrease in thigh muscle size and quadriceps femoris strength is discussed. PMID- 9353687 TI - Viscoelastic behavior of plantar flexor muscle-tendon unit at rest. AB - Muscle stretching as an exercise routine is widely used in orthopaedic and neurological rehabilitation. However, the muscle response to specific stretching parameters is still unclear. The aim of this study was to investigate the effect parameters, such as stretch velocity, stretch extent, and initial muscle-tendon resistance, on the plantar flexor response to passive movement. Eighteen healthy subjects (23-41 years) participated in this study. Five passive ankle dorsiflexions were randomly imposed at various velocities from 5 degrees/sec to 180 degrees/sec using a Kin-Com dynamometer, while unwanted activations of the soleus and tibialis anterior muscles were detected with surface electrodes. The resistive torque was averaged at -10 degrees and 0 degree of dorsiflexion. As shown by analyses of variance followed by Scheffe post hoc procedures, the resistive torque was significantly increased (p < 0.01) between 5 degrees/sec and higher velocities (60 degrees/sec or 120 degrees/sec and higher). A strong linear resistive torque-velocity relationship was also observed, as indicated by Pearson correlation coefficients of 0.92 (-10 degrees) and 0.91 (0 degree). The absolute resistive torque increment, calculated at 180 degrees/sec, was larger at 0 degree of dorsiflexion than at the -10 degrees of dorsiflexion position. Finally, subjects with larger initial plantar flexor resistance had a higher resistive torque increment (p < 0.05) at a high velocity of stretch (180 degrees/sec) than those with less initial muscle-tendon resistance. These results indicate that 1) the nonreflex resistive torque response to stretch is velocity-sensitive and 2) both a larger stretch extent and muscle initial resistance lead to greater resistive torque increments at high velocity. These observations suggest that slow and gradual stretching procedures, rather than rapid or ballistic movements, should be used, especially with stiff muscles to reduce the chance of injury from excessively high tension. PMID- 9353688 TI - Test-retest reliability of patient reports of low back pain. AB - Low back pain is, in large part, a subjective illness. Clinicians must use patient descriptions of the severity and location of low back pain and how it responds to various activities and positions to make diagnostic and treatment decisions. Therefore, it is important to understand how reliably patients describe these aspects of low back pain. The purpose of this study was to determine the test-retest reliability of a visual analogue scale measure of pain intensity, a pain drawing measure of pain location, and the pain response to activity and position questionnaire. Fifty-three subjects (28 men and 25 women) with a mean age of 54.2 years were recruited from an outpatient orthopaedic clinic. They completed the visual analogue scale, pain drawing, and pain response to activity and position questionnaire before and again immediately after seeing their physician. Thirty-three subjects also completed the visual analogue scale and pain drawing measure that evening and the next morning. Test-retest reliability of the visual analogue scale and pain drawing measure was examined using an intraclass correlation coefficient. Reliability of each item on the pain response to activity and position questionnaire was examined by calculating an unweighted Cohen's kappa. Overall, the three pain measures demonstrated fair to good test-retest reliability: 1) visual analogue scale = .66-.93, 2) pain drawing = .58-.94, and 3) pain response to activity and position questionnaire = .46-.89. The results of this study suggest that, although there is some variability in how consistently patients report various aspects of low back pain, the reliability of these pain measures is sufficient to permit their use in making clinical decisions and measuring treatment outcomes. PMID- 9353689 TI - Work-related low back injuries caused by unusual circumstances. AB - Expanding the knowledge of issues that surround work-related injuries allows for the development of more successful work accident prevention policies, treatment and rehabilitation protocols, and education programs. Specifically, clinical observation indicated that many patients did not perceive or report the circumstances of the injury as being part of their regular duties. The objective of this study was to investigate whether unusual activities or circumstances at work played any significant role in the rehabilitation of patients with disabling low back pain. Four hundred thirty-seven patients with severe disabling pain due to work-related injuries and surgery as an option for treatment were studied. The circumstances at the time of the accident were investigated, including demographic data, type of job held at the time of injury, rapport with supervisor, recent cutbacks in job force, and number of highly related injuries. Thirty-three percent of workers were injured while performing their ordinary job duties, with lifting implicated as the most common cause of injury (66%) followed by pushing/pulling (13%). In most patients (67%), the injury occurred under unusual circumstances or activities not normally described in the worker's job routine. This finding has not been previously addressed and implies that physical therapists can improve patient confidence about rehabilitation and returning to work by educating the patients about these circumstances. PMID- 9353690 TI - Optimal walking in terms of variability in step length. AB - The optimal condition in speed, step rate, and step length of human walking has been reported in terms of temporal consistency, energy cost, and attentional demand. No study, however, has been conducted on the optimal condition in terms of spatial variability of walking. This study examined whether there is an optimal walking speed with minimum intrasubject variability in step length and step width during free walk (experiment 1) and whether there is an optimal step rate with minimum step length variability during walking with imposed step rates (experiment 2). Wearing shoes with ink-applied felt squares attached to the heels, healthy students walked on a flat walkway (0.6 x 16 m) at five different speeds with a freely chosen step rate in experiment 1 and walked at three different speeds with five different step rates in experiment 2. Free walk was found to have the fewest variable errors (VEs) in step length approximately at preferred walking speed. Variable error in step width increased linearly with an increase in walking speed. Under imposed step rates, VEs in step length were the fewest when walking with step rates close to those in free walk. Our everyday walking is performed most frequently at preferred speed and/or with freely chosen step rate, thereby optimizing the consistency of gait performance. Intrasubject variability in step length may be a useful measure for evaluation of walking. PMID- 9353691 TI - Modeling of subcutaneous absorption kinetics of infusion solutions in the elderly using technetium. AB - Absorption kinetics of solutes given with the subcutaneous administration of fluids is ill-defined. The gamma emitter, technitium pertechnetate, enabled estimates of absorption rate to be estimated independently using two approaches. In the first approach, the counts remaining at the site were estimated by imaging above the subcutaneous administration site, whereas in the second approach, the plasma technetium concentration-time profiles were monitored up to 8 hr after technetium administration. Boluses of technetium pertechnetate were given both intravenously and subcutaneously on separate occasions with a multiple dosing regimen using three doses on each occasion. The disposition of technetium after i.v. administration was best described by biexponential kinetics with a Vss of 0.30 +/- 0.11 L/kg and a clearance of 30.0 +/- 13.1 ml/min. The subcutaneous absorption kinetics was best described as a single exponential process with a half-life of 18.16 +/- 3.97 min by image analysis and a half-life of 11.58 +/- 2.48 min using plasma technetium time data. The bioavailability of technetium by the subcutaneous route was estimated to be 0.96 +/- 0.12. The absorption half life showed no consistent change with the duration of the subcutaneous infusion. The amount remaining at the absorption site with time was similar when analyzed using image analysis, and plasma concentrations assuming multiexponential disposition kinetics and a first-order absorption process. Profiles of fraction remaining at the absorption site generated by deconvolution analysis, image analysis, and assumption of a constant first-order absorption process were similar. Slowing of absorption from the subcutaneous administration site is apparent after the last bolus dose in three of the subjects and can be associated with the stopping of the infusion. In a fourth subject, the retention of technetium at the subcutaneous site is more consistent with accumulation of technetium near the absorption site as a result of systemic recirculation. PMID- 9353692 TI - Pharmacokinetic-pharmacodynamic modeling of doxacurium: effect of input rate. AB - One of the basic assumptions in pharmacokinetic-pharmacodynamic modeling (PK-PD) is that drug equilibration rate constant between plasma concentration and effect (Ke0) is not changed by input rate. To test this assumption in a clinical setting, a 25 micrograms/kg i.v. dose of doxacurium was administered either by bolus injection or 10-min infusion to 15 anesthetized patients. Neuro-muscular function was monitored using train-of-four stimulation of the ulnar nerve. For the short infusion dose, arterial concentrations were measured at I-min intervals during infusion and at frequent intervals thereafter. Following the iv bolus dose, the early PK profile of doxacurium was investigated by measuring doxacurium arterial concentrations every 10 sec during the first 2 min and at frequent intervals thereafter. PK-PD modeling was performed using nonparametric approach with and without including a finite receptor concentration (Rtot) in the effect compartment. Kinetic parameters were unchanged. For the bolus and the infusion, Ke0 values were 0.053 +/- 0.006 and 0.056 +/- 0.009 min-1, respectively. Using the Rtot model, corresponding Ke0 values were 0.148 +/- 0.016 and 0.150 +/- 0.024, respectively. The relatively faster Ke0 obtained with the Rtot model is compatible with the high potency of doxacurium. Our results show that PK-PD parameters derived with either a bolus or an infusion mode of administration are equally reliable. PMID- 9353693 TI - The impact of arteriovenous concentration differences on pharmacodynamic parameter estimates. AB - In many pharmacodynamic investigations venous drug concentrations are measured and linked to effect-site concentrations by means of a traditional first-order effect-compartment model to estimate pharmacodynamic (PD) parameters. This analysis ignores the underlying physiology that arterial blood supplies both the venous sampling site and effect site. Recently, an extended effect-compartment model has been proposed that reflects physiology by postulating a first-order rate constant of equilibrium between arterial and effect-site concentrations (ke0) as well as first-order rate constant between arterial and venous concentrations (kv0). In the current paper, we evaluate the bias in PD parameter estimates if venous drug concentrations are measured and linked to effect-site concentrations by a traditional effect compartment as a function ke0, kv0, and the drug's elimination half-life (T1/2); we present an analytical solution to the differential equations characterizing the extended effect-compartment model; and we evaluate the performance of the extended effect-compartment model to estimate pharmacodynamic parameters on the basis of venous drug concentrations. Time profiles of venous drug concentrations and drug effect were simulated for a wide range of different values of the half-life of ke0 (T1/2,e0), the half-life of kv0 (T1/2,v0), and T1/2. The simulations showed that a significant bias (up to 90%) in PD parameter estimates occurred for certain values of T1/2,e0, T1/2,v0, and T1/2 if venous drug concentrations are linked to effect-site concentrations by a traditional effect-compartment model. This model misspecification is not apparent from the results of the fitting procedure. The extended effect-compartment model provided unbiased but imprecise PD parameter estimates. The extended effect compartment model was also able to analyze instances in which the venous concentrations equilibrate slower with the arterial concentrations than the effect-site concentrations, and proteresis is observed in the concentration- effect relationship. It is concluded that if the apparent T1/2 of the drug in the time period in which the decline in pharmacological effect is most pronounced is greater than 5 times T1/2,e0 and T1/2,e0 is greater than T1/2,v0 there is no need to model the underlying arteriovenous equilibrium delay. Under these conditions a traditional first-order link between venous and effect-site concentrations will yield accurate and reliable (less than 10% bias) estimates of the PD parameters such as Emax, EC50 and N. If T1/2 is less than 5 times T1/2,e0 or if T1/2,v0 is greater than T1/2,e0, the underlying arteriovenous equilibration delay needs to be taken into account in the model to obtain unbiased estimates of the PD parameters. This applies for almost all values of T1/2.v0. Arteriovenous equilibration delay can be best taken into account by measuring arterial blood concentrations. If this is not possible, the extended effect-compartment link model can be used. However, a large number of effect measurements needs to be obtained to estimate the model parameters accurately. PMID- 9353694 TI - Serum protein binding of nonsteroidal antiinflammatory drugs: a comparative study. AB - The unbound fraction in serum, fu, is a critical parameter in describing and understanding the pharmacokinetics of NSAIDs. We compared fu for 6 different NSAIDs using ultrafiltration of pooled serum at pH 7.4 and 24C. Measurements covered a wide concentration range in order to define binding affinity and number of binding sites. HPLC was used to measure drug concentrations in serum and ultrafiltrate. Direct injection of ultrafiltrate and serum (diluted 250 x) permitted quantitation down to approximately 70 nM for most of the NSAIDs, i.e., approximately 15-20 ng/ml. Assuming binding only to albumin, the data were fitted to a model of two classes of binding sites with dissociation constants K1 and K2. The lowest K1 (highest affinity) was found with flurbiprofen, 0.0658 microM, the highest with ketoprofen, 5.23 microM, an 80-fold difference. At low drug concentrations, fu becomes virtually constant and approaches a lower limit, fumin. The following fumin values were calculated: diclofenac 0.21%; fenoprofen 0.25%, flurbiprofen 0.022%, ketoprofen 0.52%, naproxen 0.039%, and tolmetin 0.37%. Thus the least bound NSAID, ketoprofen, had a value 24-fold that of the most highly bound, flurbiprofen. The NSAIDs also differed widely with regard to the extent of variation in fu within the range of therapeutic concentrations, and hence with regard to their potential as displacers of other drugs. PMID- 9353695 TI - Modeling of trough plasma bismuth concentrations. AB - Disposition pharmacokinetics of bismuth following oral dosing of ranitidine bismuth citrate are complicated and variable. An analysis of data from healthy volunteers suggests a model with three disposition compartments and first-order absorption. Patient data are pooled from 10 separate studies and consist of 1140 trough concentrations measured in 802 patients following dosing of 2 to 12 weeks duration. There are therefore insufficient data to obtain reliable parameter estimates for the full model and we use instead a much reduced model and an informative prior based on the volunteer data. Individual parameter estimates from this model can then be used to establish covariate relationships. Trough concentrations were influenced by the coadministration of clarithromycin and by creatinine clearance. A simulation study was carried out to check the validity of the estimates obtained from the reduced model. We carry out analysis via Bayesian sampling-based techniques. Throughout, we use predictive distributions for both diagnostic and inference purposes. In particular, we determine predicted distributions for the Cmax, Cmin and AUC characteristics of new individuals. PMID- 9353696 TI - Mathematical formalism for the properties of four basic models of indirect pharmacodynamic responses. AB - Four basic models for characterizing indirect pharmacodynamic responses were proposed previously and applied using differential equations. These models consider inhibition or stimulation by drug of the production or loss of mediators or response variables. This report develops partially integrated solutions for these models which allow more detailed examination of the roles of model parameters and pharmacokinetic functions in affecting the time course of drug effects. Because of the nonlinear Hill function, the solutions are represented by means of definite integrals containing kinetic and dynamic functions. These solutions allow a qualitative examination, using calculus, of how response is controlled by Dose, IC50 or SC50, Imax or Smax, and kout for drugs exhibiting monotonic or biphasic disposition. Characteristics of the response curves that were identified include shape, maximum or minimum, and changes with the above parameters and time. These relationships, together with simulation studies, provide a fundamental basis for understanding the temporal aspects of the basic indirect response models. PMID- 9353698 TI - A safe, reliable method for skin-graft coverage of the radial forearm donor site. AB - Delayed donor-site healing remains one of the most significant disadvantages of the radial forearm free flap. In an effort to decrease morbidity at the donor site, the authors adopted a closure technique that utilized the flexor digitorum sublimis (FDS) and flexor pollicis longus (FPL) muscle bellies to cover the flexor carpi radialis (FCR) tendon prior to placement of a split-thickness skin graft. While this approach eliminated tendon exposure, two patients with postoperative median-nerve compression forced a modification of this technique. The authors now detach the radial attachment of the FDS muscle and mobilize the median nerve away from the underside of the muscle, to prevent kinking of the nerve when the FDS and FPL muscle bellies are sewn together. With these modifications, the technique retains its efficacy, but with an improved margin of safety for the median nerve. PMID- 9353697 TI - Blood-flow velocity as a factor in postoperative microvascular patency. AB - The authors attempted to develop a reliable and reproducible new animal model in which the blood-flow velocity to a flap could be varied. This model was utilized to study the effects of different blood-flow velocities on the patency rate of small 1- to 2-mm vessels after common microsurgical procedures. Male Sprague Dawley rats, weighing 450 to 550 gm, were used to develop a model creating either a "high blood flow" or a "low blood flow" state by ligating the rat femoral artery, either distally or proximally, to an epigastric artery based on a groin cutaneous flap. Blood-flow velocities were measured by microvascular flowmeter, and statistical analysis was performed on the data collected. The model was next used to determine the effects of different blood-flow velocities on the patency rates of rat femoral vessels after primary anastomosis vs interpositional vein grafting. Interpositional vein grafting was subsequently repeated by a more senior microsurgeon, to determine the potential effects of increased surgical experience. The animal model was reliable, easily reproducible, and efficacious in producing two separate groups of rats with significantly different blood-flow velocities (3.98 vs. 2.14 +/- 0.5 ml/min), as was confirmed by electromagnetic flowmeter and statistical analysis. In experienced hands, decreased blood-flow velocity did not result in decreased patency rates of these small vessels after primary anastomosis, or even after vein grafting. As long as microvascular vein grafting and primary anastomosis procedures are done properly, even 1-mm vessels can tolerate significantly decreased blood-flow velocity without a decreased patency rate. Although many known factors can contribute to thrombosis and failure of anastomoses in clinical microsurgery, blood-flow velocity appears not to be a significant factor. Also described is a new, reliable animal model that can be used in small-vessel blood-flow velocity studies. PMID- 9353699 TI - Development and implementation of an extremity free-tissue-transfer database. AB - Rigorous clinical outcomes research requires accurate, complete, and standardized data. No such system is currently being used by reconstructive microsurgeons to evaluate free-tissue-transfer procedures. To facilitate collection of relevant and complete data, the authors propose a standardized format for data collection regarding these procedures. Data are collected via computer entry or scannable forms. The database includes sociodemographic, clinical, health/functional status, patient satisfaction, and resource utilization variables--the necessary components of a well-constructed clinical outcome study. Such studies will give rise to meaningful treatment algorithms for managing reconstructive problems. In addition, widespread use of a standardized databse will facilitate comparisons between practices and allow for meta-analyses. Resultant practice guidelines and microsurgery care maps will ultimately improve patient care and minimize unnecessary costs. PMID- 9353700 TI - Two-stage arterialized flow-through venous flap transfer for third-degree burn defects on the dorsum of the hand. AB - A modified, two-stage arteriovenous flow-through venous flap was designed to repair skin defects due to third-degree burns on the dorsum of the hand in four patients. Two weeks after plasty of an arteriovenous (A-V) shunt between the greater saphenous vein and dorsalis pedis artery, the arterialized flow-through venous flap was transferred using the greater saphenous vein as the pedicle. The size of the flaps utilized ranged from 7 x 13 cm to 9 x 13 cm. In three patients the entire flap survived without complication. In one patient whose flap had only one drainage vein, the flap survived with superficial necrosis of about 10 percent of the flap at the borders. During the 2 weeks after A-V shunt creation, the authors believe that microcirculation around the arterialized vein probably develops, contributing to better irrigation and thereby to flap survival. Using this two-stage procedure, it might be feasible to obtain larger grafts and to attain a higher flap survival rate. PMID- 9353701 TI - DNA amplification determines donor-cell fate in cryopreserved skin allografts. AB - Cryopreserved donor skin-cell survival was tested after allo- and isotransplantation by DNA amplification of male donor-cell genes which detects trace quantities of cells. Essentially, all cryopreserved allograft skin cells were rejected at the recipient site. Fine-haired, cryopreserved, belly skin of male BALB/c mice was transplanted onto the coarser-haired back of either female Swiss-Webster mice (allograft) or female BALB/c mice (isograft). Six weeks later, skin samples from the graft sites were tested by DNA polymerase chain reaction (PCR) amplification. Although allografts initially engrafted, more than 99.9 percent of male allograft skin cells were subsequently rejected, and gradually replaced by hairless host scar tissue. Clinically, all isografts, including hair follicles, engrafted permanently and maintained donor-cell SRY gene sequences in fine-haired graft site cells. Thus, cryopreservation maintained both the viability and antigenicity of mouse skin cells, because allografts were rejected and isografts survived. Furthermore, DNA amplification, quantified at multiple control dilutions and amplification cycles, can conclusively determine the fate of transplanted cells. PMID- 9353702 TI - Acute reconstruction of traumatic injuries of median and ulnar nerves by grafting with intercostal nerves from the rectus muscle: case reports. AB - In two cases of complicated hand and wrist injuries for which rectus flaps were used as soft-tissue coverage, intercostal nerves from the rectus muscles were harvested simultaneously to graft significant nerve defects. In one patient, a 7 cm ulnar-nerve defect was bridged with four intercostal nerves. The other patient had a 5-cm median-nerve defect repaired with three intercostal nerves. Both patients recovered either protective or normal sensation; and the patient with the median nerve injury recovered thenar muscle function. Intercostal nerves harvested with rectus muscle flaps can be the basis for acute grafting of nerve defects without using conventional nerve grafts at a traumatic site. PMID- 9353704 TI - Nasolabial fasciocutaneous free flap for cheek defects. AB - The nasolabial fasciocutaneous free flap provides another option for the reconstruction of cheek defects. The flap can offer excellent skin texture and color match, precise donor and recipient vessel size match, and the potential for a superior donor-site scar. PMID- 9353703 TI - Mucosal prelamination of a radial forearm flap for intraoral reconstruction. AB - For reconstruction of intraoral soft-tissue defects after radical resection of squamous-cell carcinomas, the microvascular jejunal patch has been a reconstructive graft option of first choice. in addition to other advantages, these jejunal grafts are able to produce mucus. In cases in which the use of jejunal grafts is contraindicated, the fasciocutaneous radial forearm flap has enlarged the spectrum of reconstructive options. A disadvantage is that mucus production will be absent, because mucosal and lining reconstruction is performed with tissue lacking mucus-providing qualities. The authors successfully prelaminated a distal radial forearm flap with buccal mucosa in five patients. Mucosal prelamination of the distal radial forearm flap enables a physiologic reconstruction with resultant mucus production, in combination with the provision of thin, pliable, and resistant flaps. The technique lowers donor-site morbidity because of the preservation of skin and subcutaneous tissue. Reconstruction with fasciomucosal, osteomyomucosal, and myomucosal flaps by this method seems feasible. PMID- 9353705 TI - What is consciousness? AB - In the past few years scientists and scholars in a variety of disciplines have been making concerted efforts to answer an ancient question, namely, How exactly do the physical processes in the brain cause consciousness? What is distinctive about the way in which modern scientists and scholars are approaching this question is that they are treating it as a scientific problem rather than a metaphysical one. This transition reflects the air of expectation in contemporary cognitive science to the effect that an empirical solution is imminent to a philosophical problem that previously was considered insoluble. Nevertheless, a recent authoritative review of the publications of such leading contemporary workers in the field as Francis Crick, Daniel Dennett, Gerald Edelman, Roger Penrose, and Israel Rosenfield has concluded that they have all failed to provide a satisfactory answer to the question (Searle 1995a). The present paper makes a psychoanalytic contribution to this interdisciplinary effort and provides an alternative answer to the question, based on Freud's conceptualization of the problem of consciousness. The paper takes a concrete example from Searle's review, reanalyses it within Freud's metapsychological frame of reference, and shows how this frame provides a radical solution to the problem. This implication of Freud's work has not hitherto been recognized and so has not received the attention it deserves. PMID- 9353706 TI - Reflections on metapsychology, theoretical coherence, hermeneutics, and biology. AB - Unable to correlate clinical findings with contemporary neurophysiology, Freud tried to anchor psychoanalysis within biology through a speculative metapsychology. Recently, epistemological objections have led to abandonment of his proposals qua scientific theory, although many still use them metaphorically. Others deny the need for any general theory of mental functions. Some theorists would espouse a hermeneutic basis for psychoanalysis, outside the boundaries of biology; they purport to confine their purview to mental contents but often use concepts based on metapsychological assumptions. Because the meanings of such contents are difficult to determine, their interpretation should be "constructed" in collaboration with analysands. By contrast, trained observers may reliably collect psychobiological data, accumulating knowledge of cognition, affectivity, communication, and the regulation of behavior--matters Freud encompassed via the economic and structural viewpoints. Hence analytic theory should be correlated with the findings of semiotics, cognitive psychology, and brain science. The hermeneutic focus on dynamics and genetics overlooks crucial data, such as the occurrence of trauma, leading to confusion about processes of pathogenesis, working through, and structural change. These and other biological phenomena (such as functional deficits and repetitive enactments) call for interventions beyond interpreting mental contents; improvement depends on learning better to process these contents. Change implies gradual establishment of alternative neural pathways; this does not automatically follow insight. Hence psychoanalysis must deal with intrapsychic phenomena beyond subjectivity. Intrapsychic conflicts represent efforts to ward off archaic mentality (primitive thought processing). Theories divorced from neurocognitive considerations encourage the theoretical fiction that analysands possess an "intact ego." PMID- 9353707 TI - A century after Freud's project: is a rapprochement between psychoanalysis and neurobiology at hand? AB - In his 1895 "Project for a Scientific Psychology" Freud attempted to construct a model of the human mind in terms of its underlying neurobiological mechanisms. In this endeavor "to furnish a psychology which shall be a natural science," Freud introduced the concepts that to this day serve as the theoretical foundation and scaffolding of psychoanalysis. As a result, however, of his ensuing disavowal of the Project, these speculations about the fundamental mechanisms that regulate affect, motivation, attention, and consciousness were relegated to the shadowy realm of "metapsychology." Nonetheless, Freud subsequently predicted that at some future date "we shall have to find a contact point with biology." It is argued that recent advances in the interdisciplinary study of emotion show that the central role played by regulatory structures and functions represents such a contact point, and that the time is right for a rapprochement between psychoanalysis and neuroscience. Current knowledge of the psychobiological mechanisms by which the right hemisphere processes social and emotional information at levels beneath conscious awareness, and by which the orbital prefrontal areas regulate affect, motivation, and bodily state, allows for a deeper understanding of the "psychic structure" described by psychoanalytic metapsychology. The dynamic properties and ontogenetic characteristics of this neurobiological system have important implications for both theoretical and clinical psychoanalysis. PMID- 9353708 TI - Psychoanalysis as the patient: high in feeling, low in energy. AB - This paper examines the increasingly important role that affect is assuming in psychoanalytic research and practice. This rise in the centrality of affect has been at the expense of an independent role for motivation and a dismissal of any energy concept. Difficulties with this affect-first approach are identified and an alternative offered that accords motivation an independent role and accommodates a useful energy concept. Research on esophageal atresia, addiction, and infant suckling are cited in support of this position. PMID- 9353709 TI - The real unconscious: psychoanalysis as a theory of consciousness. AB - Inquiries into hallucinatory wish fulfillment and the unconscious converge and, by distinguishing the concept of the unconscious in psychoanalysis from that of cognitive psychology, serve to bring out what is most essential to the psychoanalytic conception. Freud's topographical model is used to stress that the psychoanalytic unconscious can be understood only in relation to theories of consciousness and wishing. Moreover, in contrast to the cognitive conception, psychoanalysis holds that the processing of thought in the human mind is inseparable from the activity of desire. This leads to further psychoanalytic reflections on the interrelation of conscious and unconscious, wishing and thinking, and, in consequence, on transference and the mechanism of unconscious fantasy. PMID- 9353710 TI - The art and science of dream interpretation: Isakower revisited. AB - Dream imagery presents a special opportunity to lead patient and analyst through the patient's network of memories to the discovery of unconscious memories that have complicated and interfered with the patient's attempts to resolve important life problems. This is an important, perhaps indispensable, first step on the way to successful working through to solutions that are more rational and realistic than the neurotic symptoms that brought the patient to treatment in the first place. Theoretical and empirical rationales are presented for a technical approach to dreams that takes full advantage of the special opportunities that working with dream imagery provides: for deepening the psychoanalytic process and for acquainting the patient with principles of mental function applicable to aspects and phases of the analytic process that he/she will encounter as the work progresses. PMID- 9353711 TI - Does the mind fall apart in multiple personality disorder? Some proposals based on a psychoanalytic case. AB - A psychoanalytic study of some of the phenomena of multiple personality disorder (MPD), this paper takes issue with the view that a falling apart, fragmentation, or disaggregation of the mind is at the bottom of MPD's characteristic symptoms. Since first proposed by Janet in 1889, the view that ordinarily integrated parts of the mind separate from the center, accounting for the appearance of separate "selves," has prevailed among workers in this field. The close psychoanalytic study of a case of MPD suggests that, to the contrary, the appearance of multiplicity may derive from an essentially unitary but nonetheless powerful set of organizing fantasies centering on the idea that one's body and mind can be taken over and controlled by persons other than oneself. The data of the case under study suggest, further, that certain of the details of these patients' histories of childhood sexual and physical abuse may be of great importance in explaining the extraordinary organizing power of their fantasies of being occupied and controlled. In this connection, special attention is directed to the very commonly reported experiences of forced violation and the involuntary filling and emptying of their bodies during childhood. PMID- 9353712 TI - Psychiatric residency training and psychodynamic teaching. PMID- 9353713 TI - The new psychiatric texts and psychoanalysis. Essay review. PMID- 9353714 TI - Concept and early development of solid-phase peptide synthesis. AB - There are several reasons for the success of the solid-phase approach to peptide synthesis. The first is the ease of the procedure, the acceleration of the overall process, and the ability to achieve good yields of purified products. The second was the unanticipated discovery of many new biologically active peptides and the expanded need for synthetic peptides to help solve problems in virtually all disciplines of biology. In many cases, the solid-phase technique has been the method of choice. This approach, of course, does not replace the classic solution synthesis methods, but rather supplements them. The choice of techniques depends on the objectives of the synthesis. When carefully worked out, the solution methods can give high yields of highly purified products in large quantities. Many superb syntheses of active peptides have been achieved in this way. The solid-phase method has also yielded many large active peptides. It is particularly useful when large numbers of analogs, in relatively small quantities, are required as in structure-function studies on hormones, growth factors, antibiotics, and other biologically active peptides or for determining the antigenic epitopes of proteins. In addition, it has on occasion been scaled up for production of kilogram quantities. One of the unique uses of solid-phase synthesis has been the synthesis of peptide libraries. Most of the work on this new field in which thousands or millions of peptides are prepared simultaneously has been by solid-phase methods. This new technique is proving to be of great practical importance in rapid drug discovery of peptide, peptide mimetic, and nonpeptide compounds. Developments in screening methods now allow the examination of large numbers of compounds, and active products with structures unpredictable from natural product sequences are being found in this way. The properties of the solid-phase system, the changes in the chemistry, and the applications of the technique to biological problems are discussed in detail in subsequent articles of this volume. PMID- 9353715 TI - Rapid in situ neutralization protocols for Boc and Fmoc solid-phase chemistries. PMID- 9353716 TI - Cleavage methods following Boc-based solid-phase peptide synthesis. PMID- 9353717 TI - Standard Fmoc protocols. PMID- 9353718 TI - Trifluoroacetic acid cleavage and deprotection of resin-bound peptides following synthesis by Fmoc chemistry. PMID- 9353719 TI - Properties of solid supports. AB - Many supports including composite materials and functionalized surfaces are available for solid-phase synthesis. In the process of selecting the proper support it is important to consider the optimal performance during solid-phase synthesis. For most purposes the mechanically stable beaded gel resins are preferred. These resins are homogeneous, and the loading and physical and chemical properties can easily be varied. Optimal properties have been obtained by radical polymerization of end group acryloylated long-chain polyethylene glycols. However, polystyrene resins or amide bond free PEG-based resins may be more suited for general organic synthesis where reactivity of radicals, carbenes, carbanions, carbenium ions, or strong Lewis acids have to be considered. Loading of the resins can have a dramatic effect on the outcome of a synthesis and has to be considered separately for each synthesis. Synthesis of long peptides with 50 100 amino acids imposes completely different requirements on the performance, swelling, and loading than a large-scale synthesis of, for example, the pentapeptide enkephalin. Automated multiple synthesizers constructed for columns of beaded gel or composite supports are available from many suppliers. It is therefore expected that the optimization of support properties will continue in order to meet new synthetic challenges. In the synthesis for solid-phase screening of binding of biomolecules to ligands directly on the resin beads, it is an advantage if the resin is not permeable to the biomolecule so unbound molecules can easily be removed by washing. This is the case with polystyrene based resins, but they do, however, often show nonspecific adhesion of proteins owing to the hydrophobic character of the polystyrene. Modification of the functional groups of polystyrene with polyethylene glycol as spacers for synthesis of the binding ligands can increase the available ligand concentration on the bead surface and eliminate most of the nonspecific adhesion. In contrast to binding studies, solid-phase assays of enzymes require beads that are permeable to the enzyme, as the progress of reaction can be followed and the product of reaction analyzed. The available amount on the surface of the polystyrene-based beads (approximately 0.3%) is not enough for product analysis. Therefore, in the case of enzyme assays, highly swelling permeable PEG-based gel resins or functionalized surfaces of a polar and porous matrix are preferred. PMID- 9353720 TI - Coupling reagents and activation. PMID- 9353721 TI - Handles for solid-phase peptide synthesis. PMID- 9353722 TI - Solid-phase synthesis of cyclic homodetic peptides. PMID- 9353723 TI - Disulfide bond formation in peptides. AB - The goal of this review has been to present different chemical approaches for the formation of disulfide bonds in synthetic peptides and small proteins. Three general types of approaches have been described: (1) oxidation starting from the unprotected thiols; (2) oxidation starting from protected thiols; and (3) directed methods for formation of unsymmetrical disulfides. Individual or sequential disulfide-forming reactions can be carried out in solution or on a polymeric support. Overall yields and purities of products depends on protecting group combinations chosen, precise reaction conditions, and the targeted structure. Although no procedure can be guaranteed to give outstanding results for all cases, there are sufficient options available to support an optimistic view that one or more approaches can be optimized. PMID- 9353724 TI - Direct synthesis of glycosylated amino acids from carbohydrate peracetates and Fmoc amino acids: solid-phase synthesis of biomedicinally interesting glycopeptides. PMID- 9353725 TI - Synthesis of phosphopeptides using modern chemical approaches. PMID- 9353726 TI - Protein synthesis by chemical ligation of unprotected peptides in aqueous solution. PMID- 9353727 TI - Synthesis of proteins by subtiligase. AB - Application of protein engineering strategies to the redesign of the active site of subtilisin has successfully generated an efficient peptide ligase, subtiligase. The novel enzyme subtiligase has been shown to have many uses, from the total synthesis of RNase A to the semisynthesis of a variety of other proteins. Although the enzyme is in an early stage of development, it shows great promise. Subtiligase will certainly be a useful and important addition to the available strategies for the synthesis of proteins via segment condensation. PMID- 9353728 TI - Convergent solid-phase peptide synthesis. PMID- 9353729 TI - Synthetic peptide libraries. PMID- 9353730 TI - Edman sequencing as tool for characterization of synthetic peptides. AB - Sequence analysis of synthetic peptides using Edman chemistry can be very useful for the elucidation of certain types of synthetic problems, such as residue deletions and the presence of common stable derivatives, and for following the progress of the synthesis itself. However, it can also be a relatively poor technique for assessing quantitative aspects and the type and degree of adduct formation that arise from the synthetic chemistry. For these latter considerations, techniques such as mass spectrometry can often give more precise and informative data about the integrity of a synthetic peptide. Thus, sequence analysis is best applied judiciously and then used in combination with other methods. Furthermore, proper interpretation of the results of sequence analysis of synthetic peptides relies on a thorough knowledge of the sequencing process. PMID- 9353731 TI - Amino acid analysis. AB - Amino acid analysis of synthetic peptides is a robust and highly reliable analytical technique. For HPLC-purified peptides, absolute recovery accuracies of better than 5% are readily attainable. The presence of scavengers and incompletely cleaved protecting groups can have a significant impact on the quality of data that is obtained from crude samples. It should still be possible to obtain a reasonably accurate estimate of the quantity of the synthetic product, but the recovery of some amino acids can be compromised at times. When amino acid analysis is used in conjunction with mass spectrometry and a quantitative separation technique such as HPLC or CE, it can contribute significantly to the complete characterization of a synthetic peptide preparation. PMID- 9353732 TI - Analysis of synthetic peptides by high-performance liquid chromatography. PMID- 9353733 TI - Capillary electrophoresis. AB - Capillary electrophoresis is a rapid and versatile electrophoretic technique that has found several applications in the field of peptide synthesis. The high resolution and charge-based separation capabilities make it very complementary to RP-HPLC. In addition, both are essentially quantitative, which makes them ideal as purity assessment techniques. Capillary electrophoresis can be used to monitor any charge-related changes in synthetic peptides such as incomplete deprotection or chemical modifications. Capillary electrophoresis can also be very sensitive to changes in the shape or size of a peptide. However, its greatest value for the characterization of synthetic peptides is when it is used in combination with other analytical techniques such as mass spectrometry, HPLC, and amino acid analysis. PMID- 9353734 TI - Fast atom bombardment mass spectrometry of synthetic peptides. AB - Fast atom bombardment mass spectrometry plays a continuing and effective role in the rapid and efficient analysis of synthetic peptides. In this article, the basic principles of FAB-MS and FAB-MS/MS are reviewed, and the limitations and pitfalls of the method are discussed. The potential of the technique is illustrated by several selected applications. The molecular weight of a synthetic peptide can be readily and accurately determined by FAB-MS. The sensitivity of the FAB-MS method also makes it extremely useful for the evaluation of the purity of a peptide. FAB-MS/MS allows the elucidation of the primary structure of the target peptide, even in a mixture, and also permits the rapid identification of synthetic side products. Hence, FAB-MS and FAB-MS/MS aid in the unequivocal characterization of synthetic peptides. Furthermore, the information that is gained from the FAB analysis can help to unravel potential problems that may be associated with the synthesis. PMID- 9353735 TI - Analysis of peptide synthesis products by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry is an easy, rapid method for the verification of proper peptide synthesis and for the identification of most synthetic by-products. A synthesis-purification scheme has been described that uses mass analysis to (1) confirm the presence of the proper product in the crude peptide mixture, (2) guide the purification process, and (3) confirm the mass and purity of the final product. Even though many of these steps could be performed just as well with other ionization techniques, the liquid-flow characteristics of electrospray source are clearly an advantage when LC-MS is required. In addition, the ease with which fragment ions can be generated to provide structural information, even with the least sophisticated instruments, is a further advantage of ESI-MS. Although much of the operation described here was done manually, many of the steps could be automated with little additional effort (e.g., use of an autosampler). Quadrupole and ion trap instruments are widely available at present and provide the chemist with a variety of instruments from which to choose. Electrospray time-of-flight instruments will be commercially have just become available and should also provide similar results. As electrospray instruments continue to evolve, the instruments display greater performance and enhanced user-friendly interfaces, yet are lower in price and smaller in size. These features should lead to even more widespread use for the characterization of synthetic peptides. PMID- 9353736 TI - Laser desorption mass spectrometry. AB - The examples presented indicate that MALDI-MS is a useful tool for evaluating the progress of peptide synthesis at all the necessary levels: automated assembly, cleavage and deprotection chemistries, RP-HPLC analyses and purifications, and structural validation of the final product. The technique, if judiciously applied, permits the evaluation of complex peptide mixtures and often provides a semiquantitative overview. We have found that the availability of this method has enabled the provision of high-quality peptide reagents for use in the local research environment. The integration of this methodology into our peptide synthesis facility has also enabled and encouraged us to undertake more challenging synthetic problems such as phosphopeptide synthesis, peptide cyclizations, and peptide modification chemistries that would not ordinarily be offered if the laboratory lacked this technology. MALDI-MS is one of the more versatile and readily integrable mass spectrometric methods that can be incorporated into the average peptide synthesis laboratory. PMID- 9353737 TI - Protein signature analysis: a practical new approach for studying structure activity relationships in peptides and proteins. PMID- 9353738 TI - In vitro incorporation of synthetic peptides into cells. AB - This article gives a specific example of how a reversible permeabilization kit can be adapted for use with a specific peptide-based protocol. It was relatively easy to design and run the control experiments to determine the necessary adaptations. The basic procedure given with the TRANS-PORT kit is amenable to modification such that is can be used with numerous other in vitro procedures. PMID- 9353739 TI - Construction of biologically active protein molecular architecture using self assembling peptide-amphiphiles. AB - The peptide-amphiphiles described here provide a simple approach for building stable protein structural motifs using peptide head groups. One of the most intriguing features of this system is the possible formation of stable lipid films on solid substrates, or the use of the novel amphiphiles in bilayer membrane systems, where the lipid tail serves not only as a peptide structure inducing agent but also as an anchor of the functional head group in the lipid assembly. The peptide-amphiphile system potentially offers great versatility with regard to head and tail group composition and overall geometries and macromolecular structures. For building materials with molecular and cellular recognition capacity, it is essential to have a wide repertoire of tools to produce characteristic supersecondary structures at surfaces and interfaces. PMID- 9353740 TI - Chemical synthesis and nuclear magnetic resonance characterization of partially folded proteins. PMID- 9353741 TI - Multiple antigen peptide system. PMID- 9353742 TI - Relaxin. PMID- 9353743 TI - Solution nuclear magnetic resonance characterization of peptide folding. PMID- 9353744 TI - Solid-state nuclear magnetic resonance characterization of gramicidin channel structure. AB - The method of using orientational constraints derived from solid-state NMR for structural characterization of polypeptides in heterogeneous environments has now been demonstrated. A very high resolution structure has been achieved that has led to greater functional understanding of this channel. Much can be done to improve this structural technique to make it more efficient and more generally applicable. Others as well as ourselves are applying this approach to membrane proteins. Although solid-phase synthesis and specific site isotopic labeling has been essential for the development described here, one of the primary challenges is to be able to use amino acid-specific and uniform labeling of peptides and proteins by biosynthetic means for isotopic incorporation. This will allow for the study of many more proteins and significantly large proteins. Unlike solution NMR structural methods, there are no intrinsic molecular weight limitations. In fact, as the molecular weight increases the molecular motion will become less and the spectroscopic properties will improve. The major limitation will be sensitivity: as the molecular weight increases the number of moles will decrease in the samples, causing sensitivity to decrease. Advances in field strength and NMR technology help to address this problem. With larger molecules and more isotopically labeled sites resolution could also be a problem; however, the two- and three-dimensional methods demonstrated by Opella and co-workers clearly show the potential for enormous resolving power. In the 15N dimension alone it is shown that the resolution is greater than in solution NMR. Although challenges such as spectral assignments have yet to be completely solved, several approaches have been described, and the prospects are excellent for solving this and other problems facing the development of this novel approach for structural elucidation. Although there is an attempt to get away from solid-phase synthesis to solve larger molecular weight structures, peptide synthesis will continue to be important for generating single- and double-site labeled model compounds for characterizations of spin interaction tensors. Such characterizations will continue to be a very important aspect of this structural approach. PMID- 9353745 TI - Six-year study of peptide synthesis. PMID- 9353746 TI - Multiantibiotic resistance caused by active drug extrusion in Pseudomonas aeruginosa and other gram-negative bacteria. AB - All living organisms have been exposed to noxious compounds throughout their long evolutionary history and those surviving have evolved to fabricate devices that detoxicate and extrude these life threatening substances. It is likely, therefore, that all viable organisms, from bacteria to mammals, are equipped with active extrusion machinery. When bacteria are attacked by antibiotics, they use these tactics to combat the drugs and to develop resistance. Drugs extrusion machinery in Gram-negative bacteria is complex, consisting of the inner membrane transporter which acts as an energy-dependent extrusion pump; a binding protein which presumably connect both membranes; and the outer membrane exit channel. The extrusion pump assemblies are often encoded by chromosomal genes and might be expressed by mutation(s) or induced in the presence of drug(s). PMID- 9353747 TI - Antibacterial efflux systems. AB - Drug efflux, unidirectional pumping of cytotoxic drugs, is a major mechanism of antimicrobial multiresistance in bacteria. Although these efflux systems are usually chromosomally encoded, some are present on plasmids. Some of the efflux pumps are relatively well known: Emr and Acr system in Escherichia coli, whose outer membrane protein seems to be the multifunctional To1C; the mex efflux system described in Pseudomonas aeruginosa and ABC-type in Gram-negative bacteria. Also the role of efflux in Gram-positive bacteria are reviewed including Bacillus, Staphylococcus and Streptomyces. PMID- 9353748 TI - Molecular mechanisms of methicillin resistance in Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus (MRSA) strains are among the most common nosocomial pathogens. The most significant mechanism of resistance to methicillin in this-species is the acquisition of a genetic determinant (mecA gene). However, resistance seems to have a more complex molecular basis, since additional chromosomal material is involved in such resistance. Besides, overproduction of penicillinase and/or alterations in the PBPs can contribute to the formation of resistance phenotypes. Genetic and environmental factors leading to MRSA are reviewed. PMID- 9353749 TI - Molecular basis of antimicrobial resistance in non-typable Haemophilus influenzae. AB - Strains of the facultative anaerobe Haemophilus influenzae, both type b and non typable strains, are frequently multiresistant. The measurement of the antibiotic permeability of Haemophilus influenzae outer membrane (OM) shows that antibiotics can cross through the OM easily. Thus, enzymatic activity or efflux pumps could be responsible for multiresistance. An efflux system closely related to AcrAB of Escherichia coli is present in Haemophilus influenzae. However, their role in multiresistance seems irrelevant. Classical mechanisms such as plasmid exchange seems to be playing a major role in the multidrug resistance in Haemophilus influenzae. PMID- 9353750 TI - The role of outer membrane in Serratia marcescens intrinsic resistance to antibiotics. AB - Three different porins from Serratia marcescens were described. They were named Omp1, Omp2 and Omp3 and their molecular weights were 42, 40 and 39 kDa respectively. Omp2 and Omp3 showed osmoregulation and thermoregulation in a similar way to OmpC and OmpF of Escherichia coli. Permeability coefficients of the outer membrane of this species were calculated following the Zimmermann and Rosselet method. P values were similar to those obtained in Escherichia coli, which suggests that the chromosomal beta-lactamase would play a major role in the resistance of Serratia marcescens to beta-lactam antibiotics. Both MIC values and permeabilities were modified by salycilates and acetylsalycilate. Synergism between the outer membrane and the beta-lactamase was also evaluated. When bacteria grew in the presence of a beta-lactam in the medium, the beta-lactamase accounted for most of the resistance. PMID- 9353751 TI - Purification of OmpU from Vibrio cholerae classical strain 569B: evidence for the formation of large cation-selective ion-permeable channels by OmpU. AB - The outer membrane of the classical Vibrio cholerae strain 569B was isolated by sucrose density centrifugation. The simple treatment of the isolated outer membrane or the cell envelopes with different detergents allowed the purification of two outer membrane proteins, the 38 kDa OmpU and the 25 kDa OmpV. Furthermore, a 35 kDa outer membrane protein (probably the 35 kDa OmpA-like protein) was purified by two-fold treatment of the cell envelope with 2% SDS solution. A subsequent wash of the SDS-pellet with 2% Genapol buffer yielded in the 38 kDa OmpU protein, which formed SDS-resistant oligomers (66 kDa). The Genapol pellet contained OmpV. Reconstitution experiments with lipid bilayer membranes demonstrated that OmpU was a channel-forming component, whereas OmpV had a small channel-forming ability if any. The OmpU channels appeared to be large and water filled and had a single-channel conductance of about 2 nS in 1 M KCl for the monomer in a trimer, which means that they have a larger cross-section than enterobacterial porins. The channels showed rapid switching between open and closed configuration. They were slightly cation-selective, which suggests that they contain an excess of negatively charged amino groups. PMID- 9353752 TI - Beta-lactamases of Mycobacterium tuberculosis and Mycobacterium kansasii. AB - Re-emergence of infectious diseases caused by mycobacteria as well as the emergence of multiresistant strains of Mycobacterium has promoted the research on the use of beta-lactames in the treatment of such diseases. Mycobacteria produce beta-lactamases: M. tuberculosis produces a wide-spectrum beta-lactamase whose behaviour mimicks those of Gram-negative bacteria. M. kansasii produces also beta lactamase which can be inhibited by clavulanic acid. An overview on beta lactamases from both species is reported. PMID- 9353753 TI - Moderate resistance to penicillin in Neisseria meningitidis. AB - Meningococcal moderate resistance to penicillin (MICs 0.12 to 1 mg/l) was rarely reported before the 1980's in Spain. The frequency of isolation of resistant strains increased from 0.4% in 1985 to 42.6% in 1990. In the last few years, these strains have been reported in several countries, which suggests a change in the meningococcal response to penicillin. The resistance is due, at least in part, to a decreased affinity of penicillin binding protein 2 (PBP2) for penicillin. This decreased affinity has also been found in commensal Neisseriae. Population genetic studies demonstrate that recombinational events, replacing parts of the PBP2 gene by the corresponding regions of commensal species, followed by a rapid spread of the clones could be the origin of such resistant strains. PMID- 9353754 TI - Resistance to antivirals in human cytomegalovirus: mechanisms and clinical significance. AB - Long term therapies needed for managing human cytomegalovirus (HCMV) infections in immunosupressed patients provided the background for the emergence of the resistance to antivirals active against HCMV. In addition, laboratory selected mutants have also been readily achieved. Both clinical and laboratory resistant strains share the same determinants of resistance. Ganciclovir resistance may be due to a few mutations in the HCMV UL97 gene and/or viral DNA pol gene, the former being responsible for about 70% of clinical resistant isolates. Among them, V464, V594, S595 and F595 are the most frequent mutations. Because of their less extensive clinical use, much less is known about resistance to foscarnet and cidofovir (formerly, HPMPC) but in both cases, it has been associated to mutations in the DNA pol. Ganciclovir resistant strains showing DNA pol mutations are cross-resistant to cidofovir and their corresponding IC50 are normally higher than those from strains harboring only mutations at the UL97 gene. To date, foscarnet resistance seems to be independent of both ganciclovir and cidofovir resistance. PMID- 9353755 TI - Sergei I. Kuznetsov (1900-1987). The founder of Russian aquatic microbiology. PMID- 9353756 TI - Irreversibility of information and its implications for living systems. PMID- 9353757 TI - Comments on "Two generations of spore research: from father to son". PMID- 9353758 TI - Scientific writing: revising basic communication strategies. PMID- 9353759 TI - Molecular phylogenetic information on the identity of the closest living relative(s) of land vertebrates. AB - The phylogenetic position of tetrapods relative to the other two living sarcopterygian lineages (lungfishes and the coelacanth) has been subject to debate for many decades, yet remains unresolved. There are three possible alternatives for the phylogenetic relationships among these three living lineages of sarcopterygians, i.e., lungfish as living sister group of tetrapods, the coelacanth as closest living relative of tetrapods, and lungfish and coelacanth equally closely related to tetrapods. To resolve this important evolutionary question several molecular data sets have been collected in recent years, the largest being the almost complete 28S rRNA gene sequences (about 3500 bp) and the complete mitochondrial genomes of the coelacanth and a lungfish (about 16,500 bp each). Phylogenetic analyses of several molecular data sets had not provided unequivocal support for any of the three hypotheses. However, a lungfish + tetrapod or a lungfish + coelacanth clade were predominantly favored over a coelacanth + tetrapod grouping when the entire mitochondrial genomes alone or in combination with the nuclear 28S rRNA gene data were analyzed with maximum parsimony, neighbor-joining, and maximum likelihood phylogenetic methods. Also, current paleontological and morphological data seem to concur with these molecular results. Therefore the currently available molecular data seems to rule out a coelacanth + tetrapod relationship, the traditional textbook hypothesis. These tentative molecular phylogenetic results point to the inherent difficulty in resolving relationships among lineages which apparently originated in rapid succession during the Devonian. PMID- 9353760 TI - The contents of maternal testosterone in house sparrow Passer domesticus eggs vary with breeding conditions. PMID- 9353761 TI - Transposable element mobilization is not induced by heat shocks in Drosophila melanogaster. PMID- 9353762 TI - Computable scaling factor contracting or dilatating a calibrated walking distance as a function of relative motion. PMID- 9353763 TI - Neuron theory and new concepts of nervous system structure. PMID- 9353764 TI - Neurovascular relationships in the human neocortex. PMID- 9353765 TI - Serotonin-reactive neurons in the neocortex. PMID- 9353766 TI - Perforated synapses in the neocortex and their role in the reorganization of interneuron interactions in the post-ischemic period. PMID- 9353768 TI - Ultrastructural features of spinal cord sensorimotor synapses in amphibia. PMID- 9353767 TI - Projections of the ventral tegmental area of the midbrain, the substantia nigra, and the amygdaloid body in different parts of the putamen in the dog. PMID- 9353769 TI - Ultrastructure of Mauthner neurons in the living brainstem of the goldfish. PMID- 9353770 TI - Morphological differentiation of NIE-115 mouse neuroblastoma cells. PMID- 9353771 TI - Neurosecretory activity and dynamics of the lipid content of CNS neurons in Gray's mussel, a bivalve mollusk. PMID- 9353772 TI - Structural organization of receptor elements and organs of the land mollusk Pomatia elegans (Prosobranchia). PMID- 9353773 TI - Effect of nerve growth factor on the regeneration of fibers in the rat sciatic nerve. PMID- 9353774 TI - Wing muscle motoneuron function in Drosophila with mutations in the kynurenine pathway of tryptophan metabolism. PMID- 9353775 TI - Glutamate levels in the nucleus accumbens in a conditioned emotional response. PMID- 9353776 TI - Cortical mechanisms of programmed brain activity in the organization of behavioral acts in cats. PMID- 9353777 TI - Effect of a polysaccharide fraction of ginseng root on learning and memory in rats (using an active escape response as an example). PMID- 9353778 TI - Features of motor cortex neuron responses to specific stimulation in old rabbits. PMID- 9353779 TI - Role of the head of the caudate nucleus and the orbital cortex in neuronal activity of tegmentum of the midbrain in a food reflex in cats. PMID- 9353781 TI - Relationship between learning characteristics and the properties of visual objects in Rhesus macaques. AB - Behavioral studies were carried out on Rhesus macaques to investigate the relationship between the processes of learning a visual differentiation task and various properties of the stimuli, with the aim of identifying the effect on learning of a cognitive factor such as the biological significance of the visual object. Cluster analysis of a number of features of the learning process was used to distinguish visual stimuli (16 pairs) into separate compact classes of similar objects, each of which appeared to be characterized by its level of biological significance. A scheme describing the learning process, and including an assessment of the significance of sensory information, is proposed. PMID- 9353780 TI - Involvement of structures of the striato-thalamo-cortical system in an operant defensive conditioned reflex. PMID- 9353782 TI - Effect of generalized seizures on the structure of the sleep-waking cycle and the EEG in rats with an inherited predisposition to audiogenic convulsions. AB - Data are presented on the effects of generalized tonic-clonic seizures on the structure of the one-day sleep-waking cycle in Krushinskii-Molodkina (KM) rats, which have a genetic predisposition to audiogenic convulsions. Spectral and correlation analysis of EEG activity in the hippocampus, caudate nucleus, medial central nucleus of the thalamus, and in the somatosensory, visual, and auditory regions of the cortex of these animals was carried out for time intervals before and after convulsions. After seizures, rats showed a prolonged (up to 3.5 h) reduction in fast-wave sleep (FWS) with no subsequent compensatory increase in this phase in the sleep-waking cycle, while a disturbance in slow-wave sleep (SWS) was minor and short-lived (not more than 2 h). It is suggested that generalized paroxysmal attacks predominantly involve disorganization of the function of the systems regulating FWS, while the synchronizing mechanisms of the brain, responsible for SWS, are affected to a lesser extent. PMID- 9353783 TI - Neurobiological basis of creativity. PMID- 9353785 TI - Time factors in the conditioning of behavioral responses. PMID- 9353784 TI - Cholinergic dependence of a cortical neuronal mechanism that supports Pavlovian eyeblink conditioning. PMID- 9353786 TI - Neural mechanisms of autonomic responses elicited by somatic sensory stimulation. AB - All evidence introduced here indicates that, in anesthetized animals in which emotional factors have been eliminated, somatic afferent nerve stimulation can regulate various visceral functions by responses that are reflex in nature. One conclusion emerging from the evidence presented is that the effects of somatic afferent stimulation are dependent upon the particular organs and on the spinal afferent segments. When the central nervous system is intact, the responses are sometimes general, as seen in cerebral cortical blood flow, heart rate, and adrenal medullary hormonal secretion and splenic immune function, whereas sometimes they have a strong segmental organization, as seen in gastric motility and urinary vesical contractility (Fig. 8). Needless to say, in the spinalized preparation all responses are strongly segmental. The contribution of the sympathetic and parasympathetic efferent nerves to the somato-visceral reflexes depends on the organs. It is difficult for us to state specifically or to generalize upon which autonomic component, the sympathetic or parasympathetic, will dominate as the efferent path in these reflexes, because this depends on the individual organ, the site being stimulated, and the nature or mode of the stimulation. The somatically-induced reflex responses of autonomic, hormonal and immune functions demonstrated in anesthetized animals, as have been discussed herein, appear to function even during conscious states. We need further studies to evaluate the physiological meaning of these somato-autonomic reflex responses. The analysis of neural mechanisms of these reflex responses seems to be very important for clinical application to regulate visceral function by physical treatment. PMID- 9353787 TI - Role of subcortical structures in the process of conditioning. PMID- 9353788 TI - Vortrag [lecture] in St. Petersburg, October, 1994. Pavlov's dog and Benzer's fly: the genetics of higher nervous activity. PMID- 9353789 TI - Cerebral substrates of Pavlovian conditioning of discrete behavioral responses. PMID- 9353790 TI - Possible dual effect of endogenous ANP on water and sodium intake and role of AII. AB - Water intake may or may not be associated with sodium appetite. There are excitatory and inhibitory mechanisms that control these behaviors whose mediators and interactions are unclear. We investigated the effects of specific antisera against angiotensin II (AB-AII) and atrial natriuretic peptide (AB-ANP) on the induction of the two behaviors in rats deprived of water overnight or normally hydrated and submitted to intracerebroventricular (icv) microinjection of AII. AB ANP reduced water intake induced by overnight deprivation but not by icv microinjection of AII, while AB-AII reduced water intake in both situations. AB ANP and AB-AII increased saline intake in deprived animals and decreased saline intake induced by icv microinjection of AII in normally hydrated animals. The effect of AII on water and sodium intake may depend, at least in part, on an interaction with the system of ANP neurons. This peptide, in turn, may have different actions on water and sodium intake as a function of extracellular fluid conditions and of AII levels. PMID- 9353791 TI - Behavioral effects of clozapine and dopamine receptor subtypes. AB - The atypical neuroleptic clozapine (CLZ) is an extremely effective antipsychotic that produces relatively few motoric side effects. However, CLZ displays limited antagonism at the dopamine (DA) D2 receptor, the receptor commonly thought to mediate the antipsychotic activity of neuroleptics. The mechanism of action behind the efficacy of CLZ remains to be determined. Miller, Wickens and Beninger [Progr. Neurobiol., 34, 143-184 (1990)] propose a "D1 hypothesis of antipsychotic action" that may explain the antipsychotic effects of CLZ. This hypothesis is built on the interactions between D2, cholinergic and D1 mechanisms in the striatum. These authors assert that although typical neuroleptics block D2 receptors, it is through an indirect action on D1 receptors that their antipsychotic action is manifest. The extra-pyramidal side effects produced by typical neuroleptics are hypothesized to be due to an indirect action on cholinergic receptors. It is argued that the anticholinergic properties of CLZ negate the D2 (motor side effects) action of CLZ, allowing CLZ to diminish psychotic symptoms through a direct action on D1 receptors. Thus, CLZ may function as a D1 receptor antagonist in behavioral paradigms. The current paper reviews and compares the behavioral profile of CLZ to those produced by D2- and D1-selective antagonists with specific reference to unconditioned and conditioned behaviors in order to more fully evaluate the "D1 hypothesis of CLZ action". Although the actions of CLZ remain unique, they do share some striking similarities with D1 receptor antagonists especially in tests of unconditioned behavior, possibly implicating the D1 receptor in the action of this antipsychotic drug. PMID- 9353792 TI - Neuropharmacological mechanisms of nerve agent-induced seizure and neuropathology. AB - This paper proposes a three phase "model" of the neuropharmacological processes responsible for the seizures and neuropathology produced by nerve agent intoxication. Initiation and early expression of the seizures are cholinergic phenomenon; anticholinergics readily terminate seizures at this stage and no neuropathology is evident. However, if not checked, a transition phase occurs during which the neuronal excitation of the seizure per se perturbs other neurotransmitter systems: excitatory amino acid (EAA) levels increase reinforcing the seizure activity; control with anticholinergics becomes less effective; mild neuropathology is occasionally observed. With prolonged epileptiform activity the seizure enters a predominantly non-cholinergic phase: it becomes refractory to some anticholinergics; benzodiazepines and N-methyl-D-aspartate (NMDA) antagonists remain effective as anticonvulsants, but require anticholinergic co administration; mild neuropathology is evident in multiple brain regions. Excessive influx of calcium due to repeated seizure-induced depolarization and prolonged stimulation of NMDA receptors is proposed as the ultimate cause of neuropathology. The model and data indicate that rapid and aggressive management of seizures is essential to prevent neuropathology from nerve agent exposure. PMID- 9353793 TI - Sex differences in the human corpus callosum: myth or reality? AB - It has been claimed that the human corpus callosum shows sex differences, and in particular that the splenium (the posterior portion) is larger in women than in men. Data collected before 1910 from cadavers indicate that, on average, males have larger brains than females and that the average size of their corpus callosum is larger. A meta-analysis of 49 studies published since 1980 reveals no significant sex difference in the size or shape of the splenium of the corpus callosum, whether or not an appropriate adjustment is made for brain size using analysis of covariance or linear regression. It is argued that a simple ratio of corpus callosum size to whole brain size is not an appropriate way to analyse the data and can create a false impression of a sex difference in the corpus callosum. The recent studies, most of which used magnetic resonance imaging (MRI), confirm the earlier findings of larger average brain size and overall corpus callosum size for males. The widespread belief that women have a larger splenium than men and consequently think differently is untenable. Causes of and means to avoid such a false impression in future research are discussed. PMID- 9353794 TI - Tumor necrosis factor-alpha: a neuromodulator in the CNS. AB - In the central nervous system (CNS), the cytokine tumor necrosis factor-alpha (TNF alpha) is produced by both neurons and glial cells, participates in developmental modeling, and is involved in many pathophysiological conditions. There are activity-dependent expressions of TNF alpha as well as low levels of secretion in the resting state. In contrast to the conventional view of a cytotoxic effect of TNF alpha, accumulating evidence suggests a beneficial effect when TNF alpha is applied at optimal doses and at specific periods of time. The bimodal effect is related to subtypes of receptors, activation of different signal transduction pathways, and the presence of other molecules that alter the intracellular response elements such as immediate-early genes. TNF alpha may be an important neuromodulator in development of the CNS, diseases of demyelination and degeneration, and in the process of regeneration. It could induce growth promoting cytokines and neurotrophins, or it could increase the production of antiproliferative cytokines, nitric oxide, and free radicals, thereby contributing to apoptosis. PMID- 9353795 TI - Aging in the hippocampus: interrelated actions of neurotrophins and glucocorticoids. AB - Over the past two decades, evidence has been accumulating that diffusible molecules, such as growth factors and steroids hormones, play an important part in neural senescence, particularly in the hippocampus. There is also evidence that these molecules do not act as independent signals, but show interrelated regulation and cooperative control over the aging process. Here, we review some of the changes that occur in the hippocampus with age, and the influence of two classes of signaling substances: glucocorticoids and neurotrophins. We also examine the interactions between these substances and how this could influence the aging process. PMID- 9353796 TI - Motor and premotor mechanisms of licking. AB - The location, organization and anatomical connections of a central pattern generator (CPG) for licking are discussed. Anatomical and physiological studies suggest a brainstem location distributed within several subdivisions of the medullary reticular formation (RF). The involvement of widespread RF regions is evident from brainstem recording experiments in awake freely moving preparations and studies employing electrical stimulation of the frontal cortex to produce ororhythmic activity. The complex multifunctional properties of RF neurons producing licking are indicated by their activity during licking, swallowing and the rejection of an aversive gustatory stimulus. Anatomical studies place descending inputs to a brainstem CPG for licking to widely distributed areas of both the medial and lateral RF. In contrast, most projections originating from brainstem orosensory nuclei terminate primarily within the lateral RF. Because many pre-oromotor neurons appear concentrated largely in the intermediate zone of the RF (IRt), it is hypothesized that neurons from both lateral and medial sites converge within the IRt to control oromotor function. PMID- 9353798 TI - Multiple serotonin receptors: too many, not enough, or just the right number? AB - In this manuscript, current knowledge about central nervous system serotonin (5 HT) receptors is discussed with an emphasis toward describing the functional significance of the multiple 5-HT receptors. Five characteristics of 5-HT receptors, which are hypothesized to contribute to this functional significance, are discussed: (a) 5-HT has varying affinity and potency for the different receptor subtypes; (b) multiple transduction pathways are used by the different receptor subtypes; (c) receptor subtypes differ in their susceptibility to agonist-mediated desensitization/downregulation; (d) receptor subtypes interact in mediating cellular responses to the neurotransmitter; and (e) receptor subtypes respond differently to changes in the physiological environment. It is hypothesized that these characteristics of the multiple neurotransmitter receptors provide the nervous system with a capacity for coding and decoding of 5 HT-mediated neuronal transmission that could not take place with a single neurotransmitter receptor. Serotonergic regulation of female reproduction and regulation of glucocorticoid release are used to illustrate the integrative potential deriving from the existence of multiple 5-HT receptors. PMID- 9353797 TI - Neuroanatomical localization, pharmacological characterization and functions of CGRP, related peptides and their receptors. AB - Calcitonin generelated peptide (CGRP) is a neuropeptide discovered by a molecular approach over 10 years ago. More recently, islet amyloid polypeptide or amylin, and adrenomedullin were isolated from human insulinoma and pheochromocytoma respectively, and revealed between 25 and 50% sequence homology with CGRP. This review discusses findings on the anatomical distributions of CGRP mRNA, CGRP-like immunoreactivity and receptors in the central nervous system, as well as the potential physiological roles for CGRP. The anatomical distribution and biological activities of amylin and adrenomedullin are also presented. Based upon the differential biological activity of various CGRP analogs, the CGRP receptors have been classified in two major classes, namely the CGRP1 and CGRP2 subtypes. A third subtype has also been proposed (e.g. in the nucleus accumbens) as it does not share the pharmacological properties of the other two classes. The anatomical distribution and the pharmacological characteristics of amylin binding sites in the rat brain are different from those reported for CGRP but share several similarities with the salmon calcitonin receptors. The receptors identified thus far for CGRP and related peptides belong to the G protein-coupled receptor superfamily. Indeed, modulation of adenylate cyclase activity following receptor activation has been reported for CGRP, amylin and adrenomedullin. Furthermore, the binding affinity of CGRP and related peptides is modulated by nucleotides such as GTP. The cloning of various calcitonin and most recently of CGRP1 and adrenomedullin receptors was reported and revealed structural similarities but also significant differences to other members of the G protein-coupled receptors. They may thus form a new subfamily. The cloning of the amylin receptor(s) as well as of the other putative CGRP receptor subtype(s) are still awaited. Finally, a broad variety of biological activities has been described for CGRP-like peptides. These include vasodilation, nociception, glucose uptake and the stimulation of glycolysis in skeletal muscles. These effects may thus suggest their potential role and therapeutic applications in migraine, subarachnoid haemorrhage, diabetes and pain-related mechanisms, among other disorders. PMID- 9353799 TI - Sociogenic stress and rodent reproduction. AB - Social stress, which is a part of the interaction between animals, can be defined as the set of physical stresses caused specifically by the presence and actions of certain conspecifics. Dense populations are characterized by considerably increased intermale and interfemale aggressive behavior. This establishes a hierarchy which influences reproduction of the animals. Aggression of adults toward unrelated juveniles harms the physiological development of attacked young. Stress from crowding during pregnancy can affect reproductive activity even through the second generation. During postnatal development, sexual maturation of juveniles can be delayed by the presence of group-living adults. In adult females, disturbance of homeostasis after fertilization can evoke untimely termination of pregnancy. In monogamous rodents, removal of the male partner reduces the number of parturitions. In several species, recently inseminated females exposed to a strange male will lose developing embryos. Thus, sociogenic stressors are among the most important factors affecting fecundity in animals. PMID- 9353800 TI - The ventral lateral geniculate nucleus and the intergeniculate leaflet: interrelated structures in the visual and circadian systems. AB - The ventral lateral geniculate nucleus (vLGN) and the intergeniculate leaflet (IGL) are retinorecipient subcortical nuclei. This paper attempts a comprehensive summary of research on these thalamic areas, drawing on anatomical, electrophysiological, and behavioral studies. From the current perspective, the vLGN and IGL appear closely linked, in that they share many neurochemicals, projections, and physiological properties. Neurochemicals commonly reported in the vLGN and IGL are neuropeptide Y, GABA, enkephalin, and nitric oxide synthase (localized in cells) and serotonin, acetylcholine, histamine, dopamine and noradrenalin (localized in fibers). Afferent and efferent connections are also similar, with both areas commonly receiving input from the retina, locus coreuleus, and raphe, having reciprocal connections with superior colliculus, pretectum and hypothalamus, and also showing connections to zona incerta, accessory optic system, pons, the contralateral vLGN/IGL, and other thalamic nuclei. Physiological studies indicate species differences, with spectral sensitive responses common in some species, and varying populations of motion sensitive units or units linked to optokinetic stimulation. A high percentage of IGL neurons show light intensity-coding responses. Behavioral studies suggest that the vLGN and IGL play a major role in mediating non-photic phase shifts of circadian rhythms, largely via neuropeptide Y, but may also play a role in photic phase shifts and in photoperiodic responses. The vLGN and IGL may participate in two major functional systems, those controlling visuomotor responses and those controlling circadian rhythms. Future research should be directed toward further integration of these diverse findings. PMID- 9353801 TI - Pressure-flow studies: an evaluation of within-testing reproducibility--validity of the measured parameters. AB - The within-examination variation in selected test parameters in repeated pressure flow studies was determined in a retrospective study of consecutive pressure-flow examinations in 105 patients. It was further evaluated to see whether there was a systematic change in the measured parameters during retesting. To see if variation and reproducibility were influenced by the procedure of investigation, i.e., transurethral or suprapubic, patients were grouped according to the method employed. Finally, the effect of detrusor instability on the measurements was evaluated. Using the Abrams-Griffiths nomogram, patients were classified as obstructed, equivocal, or unobstructed. The test-retest variations in classification were evaluated. We found a systematic variation in Pdet.Qmax' Pdet.Open' and Qmax during testing, indicating a physiological effect of repeated pressure-flow studies resulting in a less obstructed second voiding. Accordingly, 69% of the patients who shifted group of classification during retesting shifted to a group of lesser obstruction at the second voiding. Still, 88% of patients remained in the same group of classification of bladder outlet obstruction. Within- and between-patient variations and reproducibility of the test results were not influenced by the procedure of investigation, i.e., the transurethral or suprapubic method. However, we found variations suggesting a decrease in urethral resistance and bladder contractility from test to test in the transurethral group, whereas variations suggesting an isolated decrease in bladder contractility were seen in the suprapubic group. Detrusor instability per se does not seem to cause any systematic changes during repeated testing. PMID- 9353802 TI - Comparison of a 10-mg controlled release oxybutynin tablet with a 5-mg oxybutynin tablet in urge incontinent patients. AB - Oxybutynin has long been used for the treatment of patients with detrusor overactivity and urinary urge incontinence. The short half-life of oxybutynin administered as a conventional tablet formulation or syrup requires 2-3 times daily dosage to be effective. A new controlled release (CR) tablet for once-daily administration has been developed. The efficacy and tolerability of this new controlled release tablet was compared to that of a 5-mg conventional oxybutynin tablet administered twice daily. Seventeen female incontinent patients were studied in a double-dummy crossover trial. Efficacy and tolerability were assessed by using a voiding diary, pad-weighing test, visual-analogue scale (VAS), and questionnaire. Adverse events were recorded spontaneously on a questionnaire by the patients themselves throughout the study. Serum concentrations of oxybutynin and its active metabolite N-desethyloxybutynin were studied after both a single dose and multiple dosage. There was no difference in efficacy between the two formulations. Depending on the parameters tested, the change from baseline values in a positive direction ranged from 15 to 53%. The incidence of adverse events was similar with both formulations. Oxybutynin or its metabolite showed no cumulation during the multiple dosage with a 10-mg CR tablet. The controlled release tablet formulation is as effective and as well tolerated as the conventional one, but has the advantage of only once-a-day dosage, enhancing treatment compliance. PMID- 9353803 TI - Efficacy of functional electrical stimulation in treating genuine stress incontinence: a randomized clinical trial. AB - Our objective was to determine the efficacy of functional electrical stimulation as a stand-alone therapy for female stress incontinence. The study was conducted as a prospective, double-blind, randomized controlled trial using subjective and objective outcome criteria. Patients enrolled in this study had stress incontinence consistent with International Continence Society criteria. Patients with significant pelvic prolapse or detrusor instability were excluded. Patients underwent twice-daily treatment sessions for a total of 3 months. Results were analyzed for confounding variables between the treatment and control groups. Statistical analysis was performed utilizing Fisher's exact test and the paired t test. Of the 54 patients enrolled in this study, 44 completed the program. The dropout rate was similar for both the treatment and control groups. There was no statistically significant difference between the treatment and control groups with regard to age, gravity, parity, previous antiincontinence surgery, menopausal status, or previous hysterectomy. Objective success for the treatment group was 15% and for the control group, 12.5% (NS). The subjective success for the treatment group was 25% and for the control group, 29% (NS). There was no relationship demonstrated between age, parity, previous surgery, hysterectomy, or menopausal status and the successful treatment of genuine stress incontinence with functional electrical stimulation. In this patient population, functional electrical stimulation was no more effective at improving or eliminating the symptoms of genuine stress incontinence than was the daily retention of the control probe. PMID- 9353805 TI - Maximal functional electrical stimulation in routine practice. AB - Maximal functional electrical stimulation is now an established treatment for urgency and urge incontinence. Many studies have been presented with good and consistent results. In a number of prospective studies we have previously recorded very favourable effects in stress incontinence and urge incontinence. In the present study, we have compared our previous experience with a retrospective analysis of a series of maximal functional electrical stimulation given according to a simple routine protocol and including 84 patients. The overall subjective improvement rate was 54% but the cure rate was only 5%, which is far below our experience in previous studies, as well as in others. The subjective outcome was in agreement with changes in mictrurition variables as recorded in voiding diaries. The discrepancy probably depends on a number of factors. It is suggested that the most crucial ones are patient selection, the intensity of stimulation, and the number of sessions given. It is important to realize the limitations and pitfalls of the technique when it is applied in routine practice. PMID- 9353804 TI - Relationship between urethral and vaginal pressures during pelvic muscle contraction. The Continence Program for Women Research Group. AB - Condensation is the performance of an effective pelvic muscle contraction increases urethral and vaginal pressures and is independent of demographic, clinical, and urodynamic factors. Our objective was to examine the relationship between urethral closure pressure and vaginal pressure during a pelvic muscle contraction in minimally trained women. Our secondary aim was to determine whether demographic, clinical, or urodynamic factors predict pelvic muscle contraction performance. Two hundred two women with urinary incontinence underwent multichannel urodynamic evaluation, including urethral profilometry and measurement of vaginal pressure during pelvic muscle contraction. One hundred forty-four women were diagnosed with genuine stress incontinence, 28 with detrusor instability, and 30 with mixed incontinence. Urethral and vaginal pressures correlated significantly during pelvic muscle contraction (P < or = 0.006). The ability to perform an adequate pelvic muscle contraction was independent of subject age, parity, hormonal or hysterectomy status, clinical severity, urethral support, and urethral profilometry measures (P > or = 0.42). PMID- 9353806 TI - Comparative analysis of bladder wall compliance based on cystometry and biosensor measurements during the micturition cycle of the rat. AB - The stiffness characteristics of the empty and filling bladder and the modulating influence of oxybutynin were investigated using a new biosensor system. Studies were done comparing the stiffness measured using the pressure/volume relationship with direct biosensor monitoring on male and female rats during isovolumetric contractions elicited during the cystometrogram (CMG). Bladder stiffness at zero volume, measured in vitro using the biosensor, was evaluated and compared with the stiffness of the prostate, seminal vesicles, testicles, and uterus. In 5 small anesthetized male rats, in vivo isovolumetric studies were performed and bladder stiffness was measured during the storage and contraction phase of the CMG. In 6 mature female rats, change in bladder stiffness during isovolumetric contractions was investigated following intraarterial (i.a.) administration of 0.1 and 1.0 mg/kg of oxybutynin. After the in vivo CMG was completed, an in vitro CMG was done measuring bladder stiffness. The results show that bladder stiffness, measured during the storage phase of the CMG, increased in accordance with the stretched length of bladder wall. During the in vivo CMG, bladder stiffness increased consequent to a spontaneous contraction from 10.0 +/- 1.9 g/cm to 29.9 +/- 3.0 g/cm (P < 0.005). Oxybutynin produced a significant decrease in bladder stiffness during the storage phase of the CMG, as measured using the biosensor, which was concomitant with an increase in bladder compliance derived from pressure/volume data. The incremental change in stiffness, delta K, during isovolumetric contraction decreased due to i.a. oxybutynin in accordance with a decrease of maximum detrusor pressure. These results indicate that delta K is related to the active change of viscoelastic properties of bladder smooth muscle. These findings imply that direct measurement of the stiffness of the bladder wall possesses the potential to be an objective assessment of bladder biomechanical properties and of their functional response to obstruction and pharmacological intervention. PMID- 9353807 TI - Beneficial effects of Tadenan therapy after two weeks of partial obstruction in the rabbit. AB - Tadenan (Debat Laboratories, France) is a plant extract used in Europe for the treatment of micturition disorders associated with benign prostatic hypertrophy (BPH). Prior studies demonstrated that pretreatment of rabbits with Tadenan significantly reduced the contractile dysfunction observed after 2 weeks of partial outlet obstruction. The specific aim of the present study was to determine the effect of Tadenan therapy following the creation of partial outlet obstruction. Two sets of experiments were performed: one with mild and the other with severe outlet obstruction. For both sets of experiments, male New Zealand rabbits (3-5 kg) were separated into 3 groups of 5 rabbits each. Each rabbit in groups 1 and 2 was obstructed using standard methodology. Rabbits in group 3 served as controls and did not receive any surgery. After 2 weeks, each rabbit in group 1 received Tadenan orally at 100 mg/kg/day for 3 weeks; each rabbit in group 2 received vehicle (peanut oil). After 3 weeks of treatment (5 weeks after partial outlet obstruction), rabbits were anesthetized and cystometries were performed. Immediately after cystometry, the rabbits were euthanized, the bladder rapidly removed, and 4 longitudinal strips prepared and mounted in individual baths for contractile studies. The contractile responses to field stimulation, carbachol, adenosine-5'-triphosphate (ATP), and potassium chloride (KCl) were determined, as follows: (1) Bladder mass approximately doubled in the mildly obstructed groups. Bladder mass increased significantly (3-5-fold) in the severely obstructed groups. (2) Cystometrograms from the mildly obstructed rabbits treated with peanut oil showed low compliance, whereas those of the mildly obstructed rabbits treated with Tadenan showed normal compliance. The cystometrograms of all severely obstructed rabbits showed low compliance. (3) Mild obstruction caused small but significant decreases in the contractile response to field stimulation that were reversed by Tadenan treatment. No changes were noted in response to bethanechol, ATP, and KCl stimulation. (4) Severe obstruction caused significant decreases in the response of bladder strips to field stimulation and bethanechol. Following Tadenan therapy, there was a significant improvement in the response to high-frequency field stimulation and a substantial improvement in the response to bethanechol (response equal to control). No changes were noted in response to ATP and KCl stimulation. In conclusion, Tadenan treatment reversed the bladder dysfunctions induced by mild partial outlet obstruction, and resulted in improved bladder function in the severe model of outlet obstruction. These studies are consistent with previous studies showing that Tadenan pretreatment protects the bladder against the development of contractile dysfunctions. PMID- 9353808 TI - Partial obstruction of the rat urinary bladder: effects on mitochondria and mitochondrial glucose metabolism in detrusor smooth muscle cells. AB - The aim of the present study was to measure mitochondrial function in the obstructed rat bladder, which does not seem to have impaired contractility in vivo. The animals were unoperated control rats and rats with a 12-day partial urinary outlet obstruction. The obstruction increased bladder weight 3-fold. The relative volume (3.5%) in the detrusor smooth muscle cells composed of mitochondria was unaffected by obstruction. Obstruction did not affect malate dehydrogenase and citrate synthase activity when expressed relative to unit bladder weight. There was, however, a significant decrease in enzyme activity when expressed relative to protein content. This was due to an increased relative protein content in the obstructed bladders. Total enzyme activities per bladder were increased. Oxygen consumption rates in maximally activated intact control and obstructed preparations in other studies corresponded to a citrate synthase (rate-limiting enzyme) activity only 10% of the maximal enzyme activity found in the present study. We conclude that there is a considerable safety margin in mitochondrial function in intact rat detrusor muscle cells, and that detrusor smooth muscle cells can hypertrophy without any impaired mitochondrial function. PMID- 9353809 TI - The ContiNet of the International Continence Society. AB - This is an account of the International Continence Society's ContiNet--the web server linking up continence organisations worldwide with provision to upload or download vast data stores of information on continence via e-mail, FTP, mailing lists, and special tools to seek information using "search engines." Special communication devices using internet voice/phone mail and real-time "text" or "voice" chats permit conversation globally over normal phone lines linked to the Net at local telephone rates. Special features of ContiNet include announcements of upcoming conventions, information for professionals and laypeople, and the capability to conduct research via the net and conduct consultations and discussions via newsgroups. In-built devices requiring special IDs and passwords permit privacy and security for users. Simple instructions are provided on how to get your PC up and running and get connected to fellow members of ICS, link up with national continence societies, or simply surf for professional enrichment and leisure. With the advent of advanced multimedia capabilities, the current poor quality videoconferencing on the Net will be replaced by excellent videophones by 1998. PMID- 9353810 TI - Comment on "Interference pattern in perineal muscles: a quantitative electromyographic study in patients before and after transurethral surgery of the prostate" in Neurourol. Urodyn. 16:101-109, 1997. PMID- 9353811 TI - Latex allergy in health care workers. AB - Since the 1980s, universal precautions have been instituted to protect against bloodborne infections. However, protective equipment can cause allergic sensitization, with the potential for severe and even life-threatening reactions. Latex gloves, in particular, have been problematic. PMID- 9353812 TI - Health effects of anesthetic gases. AB - There are no national regulatory standards from the U.S. Occupational Safety and Health Administration for anesthetic gases. Moreover, there is little data to confirm that current scavenging and exhaust ventilation systems reduce exposure below recommended limits. Operating theater personnel and workers in postanesthesia units are especially at risk. PMID- 9353813 TI - Engineering controls for abating airborne biologic, chemical, and physical contaminants in health care workplaces. AB - Airborne hazards present special challenges due to nonuniform release into the environment, inadequate or nonexistent measuring capability, and lack of established exposure limits. However, some general guidelines are available. PMID- 9353814 TI - Respiratory protection in the health care setting. AB - Respiratory protection is of increased importance due to the resurgence of tuberculosis. This chapter examines protective devices and regulations and explains how a program can be designed to minimize workplace hazards. Of particular value is a table detailing 12 high-efficiency particulate air respirators that meet criteria set by the Centers for Disease Control and the National Institute of Occupational Safety and Health. PMID- 9353815 TI - Chemical hazards in health care workers. AB - A comprehensive occupational health program is essential in health care settings to minimize the risk of occupational injury and illness in chemically exposed workers. Careful exposure assessment is the framework on which such a program is built. Medical surveillance provides an additional check by allowing earlier identification of at-risk workers. Regular analyses of these data are needed to increase knowledge regarding occupational hazards for health care workers. Correlation of adverse health effects detected in medical surveillance with exposure level is a powerful, although underutilized, tool for advancing occupational health. Worker training is the other critical element in an effective occupational health program. Employees are better able to comply with workplace rules designed to protect health and safety if they understand their rationale. PMID- 9353816 TI - Hazardous drugs. AB - Engineering controls and personal protective equipment are crucial for health care professionals working in situations where cytotoxic agents, pharmaceuticals such as ribavirin and pentamidine, and other potentially hazardous drugs are present. Plans for exposure control, spill clean-up, and medical surveillance must be instituted to protect the employee. PMID- 9353817 TI - Ergonomics programs for health care organizations. AB - There are many reasons to start an ergonomics program in a health care facility. The cost of injuries and the quality of service are two important justifications. Across all industries successful ergonomics programs have similar elements: management commitment, employee involvement, worksite analysis, hazard prevention and control, medical management, training, and evaluation. While injuries and quality are good ways to justify starting a program, the program ultimately must support the objectives of the administration. Therefore, the program must be planned in advance and measures used to show that the progress in ergonomics is consistent with the goals of the company. As the program expands by individual job and by department, it is important to remember that the goal of the program is to weave the knowledge and application of ergonomics into the fabric of the company culture. PMID- 9353818 TI - Violence prevention at the health care site. AB - Dr. Felton discusses risk factors for violence in the health care setting and recommends worksite analysis procedures, hazard prevention controls, and training and education programs for the greatest protection possible. An appendix offers three contacts for additional materials on devising preventive programs. PMID- 9353819 TI - Transmission and control of bloodborne viral hepatitis in health care workers. AB - Although the incidence of clinical HBV has declined as a result of infection control measures and vaccine-induced immunity, the prevalence of patients who are HBsAg-positive has increased. HCWs who are exposed to the blood and body fluids of patients should be required to receive hepatitis B vaccine. However, there is no vaccine against HCV, the most prevalent bloodborne pathogen in the health care setting. It therefore is critical for health care workers to encourage the development and assessment of effective preventive and control strategies, including the design and use of safe devices, targeted interventions based on occupation-specific hazards, and surveillance and analysis of exposures in the health care setting. PMID- 9353821 TI - Human immunodeficiency virus in the health care setting. AB - Dr. Wheeler reviews the mechanisms of HIV infection and discusses procedures and situations that place health professionals at risk. Prevention of HIV transmission, institutional management of occupational exposure, and postexposure prophylaxis are addressed. PMID- 9353820 TI - Vaccine-preventable diseases in health care. AB - It cannot be overemphasized that good preventive measures such as universal precautions, good hand-washing techniques, and appropriate personal protection is the mainstay of reducing the risk of nosocomial infections to other patients and HCWs. Vaccines that are safe and effective in reducing risks of nosocomial infections must be promoted through educational campaigns and made easily available to all employees, including those on night and weekend shifts. Extended hours and onsite vaccination programs help to increase acceptance of vaccines at certain times, such as during the annual influenza vaccine campaign. When more than one vaccine is recommended, consideration must be given to an appropriate vaccine schedule. Table 2 lists the interactions and contraindications of the vaccines most commonly administered to health care workers. Certain vaccines, such as rubella, should be required for susceptible employees when working in an area of a health care facility where exposure and consequent transmission to patients could have life-threatening consequences. PMID- 9353822 TI - Home care workers: injury prevention through risk factor reduction. AB - Home health care professionals work in a nonstandard and unpredictable environment for which few controls are available. The professional must cope with a residence's existing access, cleanliness, facilities, and other occupants (including pets), among other factors, and these vary between homes. This chapter suggests interventions that can reduce risks to employees, patients, and family members. PMID- 9353823 TI - Tuberculosis in the health care industry. AB - Between 1986 and 1992, a resurgence of tuberculosis in the United States made this disease once again a significant risk to health care workers. Traditional approaches remain viable methods of reducing present-day hazards. PMID- 9353824 TI - Detection of terminal deoxynucleotidyl transferase (TdT) in nonhematopoietic small round cell tumors of children. AB - In the differential diagnosis of small round cell tumors (SRCTs), terminal deoxynucleotidyl transferase (TdT) is often used as a marker for lymphoblastic lymphomas and leukemias. However, the specificity of TdT using the avidin-biotin immunoperoxidase (ABC) method is not well documented. To address this issue, we stained paraffin-embedded biopsy specimens of 64 cases of childhood SRCTs using the ABC method with anti-TdT. For any TdT-positive tumors, an additional antibody panel for lymphoid markers was applied. Two patterns of TdT positivity were observed: (1) tumor specific, consisting of strong to moderate nuclear staining, and (2) scattered positive lymphoid cells, usually in a perivascular location and expressing T-cell markers. Analysis showed that 7 of 10 medulloblastomas stained with TdT in a tumor-specific pattern (4 cases moderately to strongly positive in 75-100% of tumor cells, 3 cases weakly to moderately positive in 25-50% of cells). Also, 1 of 19 rhabdomyosarcomas and 1 of 8 Ewing's sarcomas showed moderate to strong tumor-specific TdT staining in 100 and 10% of cells, respectively. Scattered TdT-positive lymphoid cells were observed in 27% of these 64 SRCTs. These findings emphasize that TdT positivity should not be relied upon exclusively for making a diagnosis of lymphoblastic leukemia or lymphoma or ruling out other SRCTs. PMID- 9353825 TI - Histopathologic findings in the lymphoid and reticuloendothelial system in pediatric HIV infection: a postmortem study. AB - The present report describes the histopathological features of lymphoreticular tissues in 29 pediatric autopsies of human immunodeficiency virus (HIV)-infected patients. Mean age for the whole group was 1.77 years; 68.9% and 62% of the cases were 2 years old or less and 1 year old or less at the time of death, respectively. Twenty-one cases were categorized as acquired immunodeficiency syndrome (AIDS) and the rest included seven HIV-positive newborns and infants and two infants belonging to a high-risk group. The thymus (24 cases) showed severe lymphoid depletion (atrophy) in 16 (66.6%) cases, microcystic transformation of Hassall's corpuscles (HCs) in 4, calcified HCs in 3, absence of HCs in 3, and plasmacytic infiltrates and Warthin-Finkeldey-type multinucleated giant cells (also found in lymph nodes and bowel lymphoid aggregates in the same case) in 1. Lymph nodes (25 cases) revealed extensive lymphocyte depletion (68%); selective follicular (2 cases) or paracortical (3 cases) atrophy; hemophagocytosis (44%); some type of hyperplasia (plasmacytosis, enlarged follicles) in 5 cases; some type of lymphadenitis (12 cases), 5 cases of which were due to opportunistic infections (cytomegalovirus, 2; histoplasmosis, cryptococcosis, Mycobacterium avium-intracellulare, 1 each). Main findings in the spleen (28 cases) were extensive lymphocyte depletion (10 cases), limited to the white pulp in 4 and including the red pulp in 7; some type of lymphoid hyperplasia (limited to white pulp in 6 cases and involving the red pulp in 5); hemophagocytosis (7 cases); and foci exhibiting a peculiar arrangement of spindle-shaped cells combined with capillaries, plasma cells, and occasionally siderophages in 11. These we have termed kaposiform areas due to the resemblance to the so-called inflammatory variant of Kaposi's sarcoma. This pattern was also recognized in lymph nodes of two cases. Although atrophy was the main theme, cases with hyperplasia were also noticed. The possible relationship, if it exists at all, between kaposiform areas and Kaposi's sarcoma remains to be established. No tumor was found in this series. No specific histopathologic pattern of lymphoid tissues atributable to HIV emerged form this study aside from kaposiform areas, a microscopic feature not previously reported in this circumstance in pediatrics. PMID- 9353826 TI - Hepatoblastoma: the Indiana experience with preoperative chemotherapy for inoperable tumors; clinicopathological considerations. AB - Analysis of the prognostic importance of various clinicopathological parameters in 17 hepatoblastomas (HBs) confirmed the utility of preoperative chemotherapy to convert inoperable to resectable tumors. There was no significant survival advantage for patients who underwent initial tumor resection compared with those resected following chemotherapy, although complete resection, with or without prior chemotherapy, was critical for cure. Young age was associated with better survival but did not correlate with histologic subtype or clinical stage. A relationship between low initial alpha-fetoprotein (AFP) level and tumor resectability was noted, perhaps related to tumor size, but tumor location was of greater importance than size in determining resectability. Neither the mean proportions of fetal and embryonal epithelium, nor their mitotic activity, nor the presence of vascular invasion in the prechemotherapy biopsy specimens was predictive of outcome, but the low mitotic activity of the fetal component correlated with ultimate resectability. On the other hand, although complete resection was necessary for survival, histologic examination of postchemotherapy specimens had additional predictive value; the presence of vascular invasion, the amount of viable mesenchyme, the extent of tumor necrosis, the proportion of embryonal epithelium, and the mitotic activity of the epithelial component in postchemotherapy resection specimens were each predictive of outcome. Although the presence of osteoid was not predictive, both the proportion of HBs that contained osteoid and the extent of mature mesenchymal tissues within individual HBs were increased by chemotherapy, suggesting that maturation of previously immature clones had been induced. We conclude that although complete resectability remains the fundamental goal of therapy, evaluation of the clinicopathologic characteristics that we have found to be predictive of outcome may permit tailoring of therapeutic regimens to individual patients; those whose tumors are deemed likely to respond well may require less toxic preoperative chemotherapy, and those deemed likely to progress in spite of complete resection may be considered for more aggressive postoperative regimens. PMID- 9353827 TI - Heterogeneity of MYCN amplification in a child with stroma-rich neuroblastoma (ganglioneuroblastoma). AB - Amplification of MYCN portends rapid tumor progression and poor prognosis in neuroblastoma. MYCN copy number has been described as homogeneous within a tumor and congruent in primary tumor and metastasis. We report a child with stage III favorable histology stroma-rich neuroblastoma (ganglioneuroblastoma) and a poor outcome with an apparent change in MYCN gene amplification by Southern blot. Initial biopsy revealed a ganglioneuroblastoma with predominance of differentiating cells designated as neuroblastoma, stroma-rich, intermixed (Shimada). Southern blot failed to demonstrate MYCN gene amplification. After front-line chemotherapy failed, a total resection was performed. In this specimen, Southern blot demonstrated MYCN amplification (15-20 copies) in the undifferentiated component and no amplification in the differentiated. Fluorescence in situ hybridization (FISH) analysis performed retrospectively on both tumor biopsies demonstrated MYCN amplification in the undifferentiated sections of both tumor specimens but not in the differentiated ones. This is the first well-documented case report of heterogeneous MYCN amplification in a child with neuroblastoma. Because key therapeutic decisions are based on the presence of MYCN amplification, physicians diagnosing and treating children with neuroblastoma need to be aware of the possibility that MYCN amplification may be heterogeneous within a tumor and may be missed using techniques based on pooled DNA such as Southern blotting. FISH may be a preferable method for determining MYCN amplification. PMID- 9353828 TI - Eosinophilic infiltration of the enteric neural plexuses in Hirschsprung's disease. AB - Inflammatory infiltrations of the enteric plexuses are uncommon and are usually lymphoplasmacytic. Within the past 15 years, nine pediatric cases in which a predominantly eosinophilic infiltrate of the gastrointestinal wall with a predilection for the myenteric (Auerbach's) and deep submucosal (Henle's) plexuses were seen at our institution. Two were neonates without gastrointestinal abnormalities who expired shortly after birth. Seven were patients with short segment Hirsch-sprung's disease. There was a mild increase in mucosal eosinophils in the overlying mucosa and only one patient had peripheral eosinophilia. Follow up data obtained 1 month to 7 1/2 years after biopsy revealed no development of inflammatory bowel disease, connective tissue disease, malignancy, allergic disorder, or intestinal dysmotility. The proximal location of the infiltrate suggests that it may represent a secondary finding rather than a primary cause of aganglionosis. PMID- 9353829 TI - Analysis of ploidy and proliferative activity in childhood non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD). AB - We have performed DNA analysis by means of fluorescence-activated cell cytometry on paraffin-embedded tissue from the diagnostic biopsy specimens in 40 cases of non-Hodgkin's lymphoma (NHL) and 25 of Hodgkin's disease (HD) and from 50 normal tonsils as controls. For HD cases, aneuploidy was found in 7 of 25 (28%), a higher proportion than in two previous studies of mainly adult patients. Diploid tumors showed S-phase fractions (SPFs) similar to those of controls. In the NHL cases aneuploidy was found in 12 of 40 (30%) with no significant association with site, stage, histopathology, immunophenotype, or prognosis. SPFs were highest in abdominal and chest primary sites but were not related to stage. Burkitt's lymphomas had the highest SPFs relative to lymphoblastic (P < .01) and centroblastic lymphomas (P < .05). Significantly higher SPFs were found in B cell than in T cell tumors (P < .001). There was considerable heterogeneity for SPFs within each NHL subgroup. Survival was worse at 5 years for those with high SPFs compared with those with normal SPFs (P = .04). These results suggest that tumor DNA analysis may be useful in the evaluation of children with NHL. Larger studies are needed to define its role as an independent prognostic variable. PMID- 9353830 TI - Congenital parvovirus infection. AB - Congenital parvovirus infection was diagnosed in two liveborn premature infants born at 24 and 35 weeks of gestational age. The illnesses were associated with placentomegaly, petechial rash, edema, hepatomegaly, anemia and thrombocytopenia, respiratory insufficiency, and death at 5 and 6 days of age. The syndromes exhibited by these cases shared common but nonspecific features with other life threatening congenital infections. Serological studies in one case supported the diagnosis of parvoviral infection. Postmortem examination of both revealed nuclear inclusions in erythroid precursor cells characteristic of parvovirus infection. Use of the polymerase chain reaction confirmed the presence of parvovirus DNA in one of the cases. Intrauterine parvovirus B19 infection is most commonly associated with hydrops fetalis, "transient" hydrops, or a favorable outcome in infants found to be viremic after birth. These and previously reported examples of congenital B19 disease exemplify an exceptional form of human parvovirus infection. PMID- 9353831 TI - Immunohistochemistry of medulloepithelioma and neural tube. AB - Immunohistochemistry profiles of medulloepithelioma (from two 2 1/2-year-old girls who had cerebral medulloepitheliomas and a 35-week postconceptional female infant with congenital posterior fossa tumor) and neural tube are compared. Microscopically, the tumors contained a medulloepitheliomatous component, manifested as tubular epithelial structures lined by pseudostratified columnar epithelium delineated by well-defined basement membranes. In all cases, glial and neuronal differentiation were noted to differing extents. The medulloepitheliomatous components did not exhibit glial fibrillary acidic protein, neuron-specific enolase, or S-100 protein reactivity. Neurofilament, cytokeratin, and epithelial membrane antigen were focally present in one case. Extensive nestin immunopositivity was confined to the basal cell layer of the epithelium, leaving the luminal surface unreactive or slightly reactive. These cells also displayed a reactivity to vimentin and to microtubule-associated protein type 5 similar to that of cells of the primitive neural tube. The similarity between the immunohistochemical profile of medulloepithelioma and that of neural tube epithelium suggests a possible reexpression of that component of the genome responsible for neural tube growth and differentiation in medulloepithelioma. PMID- 9353832 TI - Detection of respiratory syncytial virus nucleic acid in archival postmortem tissue from infants. AB - Archival lung tissue from 99 cases of sudden infant death syndrome (SIDS) and from 58 matched comparison cases with known causes of death was studied. Sections were examined by in situ hybridization (ISH) using a cocktail of three synthetic oligonucleotides with sequences chosen from the published sequence of the nucleoprotein gene of respiratory syncytial virus (RS virus). The oligonucleotides were end-labelled with dinitrophenyl (DNP) or digoxigenin (DIG) and hybrids were detected immunocytochemically. RS virus nucleic acid was detected in 24 cases of SIDS (24%) and in 11 (19%) of the comparison group. Specificity was confirmed using a DIG-labeled cloned probe covering the whole of the nucleoprotein gene sequence. With one exception, the same results were obtained. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to confirm the specificity of these results. When matched for age and month and year of death, 76 SIDS cases and 38 controls could be compared. Twenty-one SIDS cases (27.6%) and seven comparison cases (18.4%) contained detectable RS virus sequences by ISH, with a higher detection rate in winter in both groups. The differences were not significant and reflected the seasonal pattern of RS virus infection in the community rather than a causal relationship of RS virus with SIDS. Detection of RS viral mRNA through the summer months suggests that persistence is possible. PMID- 9353833 TI - Craniopagus parasiticus: a case illustrating its relationship to craniopagus conjoined twinning. AB - A case of craniopagus parasiticus is described in which the parasitic twin is more fully developed anatomically than in any of the previous reports. Somatic and placental vascular anastomoses between the twins and hypoplasia of the umbilical cord of the parasite were also observed. These findings support the hypothesis that craniopagus parasiticus results from compromise of the blood supply to one of a pair of craniopagus conjoined twins. PMID- 9353834 TI - Intracardiac epithelial cyst associated with esophageal atresia. AB - We describe an intracardiac epithelial cyst in association with esophageal atresia. The case is unusual in that the cyst was symptomatic and ultimately fatal. In addition, there was no other cardiac anomaly, although a range of extrathoracic malformations was present. There are three types of intracardiac epithelial cysts: congenital polycystic tumor of the atrioventricular node, a cyst as part of a teratoma, and, as in this case, a gross cyst. All of them are very rare. The association of a cardiac cyst and esophageal atresia in our case supports the theory that intracardiac cysts are derived from misplaced foregut. PMID- 9353835 TI - Pulmonary endodermal tumor resembling fetal lung: report of a case in a 14-year old girl. AB - This report describes the clinical and histologic features of a pulmonary tumor in a 14-year-old girl that is most consistent with a rare entity described in the literature as "pulmonary endodermal tumor resembling fetal lung" (PET). This tumor is composed of glycogen-rich columnar cells forming complex glands with focal festooning and mitotic activity, admixed with solid "morules" of cells with eosinophilic cytoplasm and focal nuclear clearing. Patchy tumor necrosis and a bland stroma were also present. Immunoreactivity for carcinoembryonic antigen (CEA), alpha 1-antichymotrypsin, and 12E7 was present in glandular cells and for human chorionic gondatropin (HCG), alpha 1-antichymotrypsin, and 12E7 in morular cells. Ultrastructural features are those of an epithelial tumor. Related entities have been called "pulmonary blastoma lacking sarcomatous elements" and "adenocarcinoma of fetal lung type." Most cases of PET have occurred in adults, and the histologic features thought to have prognostic significance in small published series are applied to our case, in which the patient remains well and without evidence of tumor recurrence or metastasis for 28 months following local resection as the sole treatment. PMID- 9353836 TI - Congenital alveolar capillary dysplasia: rare cause of persistent pulmonary hypertension. AB - We report on a rare case of fatal congenital alveolar capillary dysplasia. The newborn boy of a 37 weeks' normal gestation suffered from persistent pulmonary hypertension without any cardiovascular malformation and died at the age of 4 weeks despite intensive treatment. The autopsy tissue was examined histologically, immunohistochemically, and ultrastructurally. Moreover, a three dimensional tissue reconstruction based on serial sections was performed comparing the affected lung with normal lung tissue. We observed a unique pattern of pulmonary dysplasia: An extreme decrease of capillaries was localized centrally within thickened intra-acinar septa instead of capillaries intensely neighboring pneumocytes; ectatic veins normally running in the interlobular septa were found to accompany intralobular bronchovascular bundles, denying a clear distinction between pulmonary and bronchial veins; small muscular pulmonary arteries extended to the precapillary level and type 2 pneumocytes exceeded by far the type 1 pneumocytes, inverting the normal ratio. In summary, alveolar capillary dysplasia is assumed to be a primary capillary disorder of unknown origin, which possibly involves the regular differentiation of pneumocytes, according to the close alveolocapillary relationship during pulmonary ontogenesis. We consider the venous alterations as being part of the dysplasia, whereas the arterial phenomena might occur secondarily. Recent reports on affected siblings suggest a genetic component of pathogenesis. PMID- 9353837 TI - Neonatal meningitis and multiple brain abscesses due to Citrobacter diversus. AB - We report a fatal sporadic case of neonatal Citrobacter diversus meningitis with rapid clinical progression. At autopsy, multiple brain abscesses and abscesses in the spinal cord had complicated purulent meningitis. Septic omphalitis was identified as the most likely portal of entry. PMID- 9353838 TI - Pluripotential melanoblastoma: authors' response. PMID- 9353839 TI - Diagnosis of chorioamnionitis. PMID- 9353841 TI - Embolism of fetal brain tissue to the lungs. PMID- 9353842 TI - Human eosinophils are not pigment-laden macrophages in Pseudomelanosis duodeni. PMID- 9353843 TI - Aspirin for the second hundred years: new uses for an old drug. AB - The history of aspirin can be traced back to ancient Egypt where extract of willow bark was used to treat inflammation. The active component of the extract was identified as the glucoside of salicylic alcohol. The severe gastric side effects associated with the use of sodium salicylate prompted the synthesis of the o-acetyl-derivative as a possible pro-drug. In fact, acetylsalicylic acid was antiinflammatory, analgesic and antipyretic but also ulcerogenic to the stomach. Acetylsalicylic acid was synthesized one hundred years ago, and was mass-produced under the commercial name of 'Aspirin' (Dreser, 1899) by the German company Bayer for the treatment of fever and rheumatism. PMID- 9353844 TI - Lead binding to delta-aminolevulinic acid dehydratase (ALAD) in human erythrocytes. AB - Over 99% of the lead present in blood is usually found in erythrocytes. To investigate the nature of this selective accumulation of lead in erythrocytes, the specific binding of lead to proteins in human erythrocytes was studied using liquid chromatography coupled to inductively coupled plasma mass spectrometry (LC ICP-MS). The principal lead-binding protein had a mass of approximately 240 kDa, and adsorption to specific antibodies showed that protein was delta aminolevulinic acid dehydratase (ALAD). Thus, the previous notion that lead in erythrocytes was bound primarily to haemoglobin has to be revised. Furthermore, in lead-exposed workers, the percentage of lead bound to ALAD was influenced by a common polymorphism in the ALAD gene. Specifically, in seven carriers of the ALAD2 allele, 84% of the protein-bound lead recovered was bound to ALAD compared to 81% in seven homozygotes for the ALAD1 allele whose erythrocytes were matched for blood-lead concentration. The small difference was statistically significant in Wilcoxon matched-pairs signed-rank test (P = 0.03). No ALAD allele-specific difference in ALAD-bound lead was found among 20 unexposed controls. Perhaps the difference in ALAD-bound lead can provide an explanation for the previously reported finding of higher blood-lead levels among carriers of the ALAD2 allele than among ALAD1 homozygotes in lead-exposed populations. PMID- 9353845 TI - Alterations of physostigmine-induced yawning by chronic lithium administration in rats. AB - The effect of chronic lithium pretreatment on physostigmine-induced yawning was investigated in male rats. Intraperitoneal administration of physostigmine to rats induced yawning in a biphasic manner. However the maximum response was obtained by 0.2 mg/kg of the drug. Intracerebroventricular administrations of a putative M1 and M2 muscarinic receptor antagonists, pirenzepine and methoctramine decreased physostigmine-induced yawning. Intraperitoneal administration of a non selective muscarinic receptor antagonist, atropine, also decreased the physostigmine-induced yawning significantly. Chronic lithium pretreatment (30 days) reduced yawning induced by physostigmine. The inhibitory effect of pirenzepine, methoctramine and atropine on physostigmine-induced yawning increased in rats pretreated with chronic lithium. These findings indicate that yawning is induced by a central cholinergic mechanism and that chronic pretreatment of lithium may interact with the cholinergic-induced behaviour. PMID- 9353846 TI - Glutathione pretreatment lessens the acute liver injury induced by carbon tetrachloride. AB - We explored the protective activity of glutathione (GSH) in a rat model of carbon tetrachloride-induced acute liver damage. Histological examination of livers from GSH-pretreated rats revealed minor damage, confirmed by biochemical parameters of liver cell necrosis evaluated both 24 and 48 hr after hepatotoxin delivery. In addition we quantified changes in hepatic steady-state levels of albumin, type I procollagen, transforming growth factor-beta 1 and tumour necrosis factor-alpha mRNAs. Even at the molecular level and above all for the albumin gene, it appears that GSH lessens the effect of the hepatotoxin, however the protection of the thiol is restricted to the first 24 hrs and is almost totally exhausted after 48 hr. Since, only 24 hr after CC14 delivery, GSH abundance determined in erythrocytes and liver is almost equal in the controls and in the GSH injected rats, but significantly higher than in the only intoxicated animals (P < 0.05, intraerythrocyte content), we conclude that the thiol pretreatment exerts an effective but transient protection. PMID- 9353848 TI - Protection by 4-methylthiobenzoic acid against cisplatin-induced ototoxicity: antioxidant system. AB - This study was undertaken in order to determine the changes in auditory brainstem evoked responses relationship with the changes in the levels of GSH, lipid peroxidation and antioxidant enzymes activity in cisplatin-induced ototoxicity and otoprotection by 4-methylthiobenzoic acid (MTBA). Male Wistar rats in different groups were treated as follows: 1) saline control; 2) cisplatin (16 mg/kg, intraperitoneally); 3) MTBA (250 mg/kg, intraperitoneally), and 4) cisplatin plus MTBA. Post-treatment auditory brainstem-evoked responses were performed after three days and the rats were sacrificed and cochleae harvested. The cochleae were analyzed for glutathione (GSH), antioxidant enzyme activity, and malondialdehyde levels. The cisplatin injected rats showed a threshold elevation of 31.9 +/- 16.0 dB above the pretreatment thresholds using click stimulus. Rats treated with MTBA plus cisplatin did not show significant elevation of hearing threshold. Cisplatin plus MTBA administration showed a higher levels of cochlear GSH (5.59 +/- 0.35 nmoles/mg protein) compared to cisplatin alone (4.46 +/- 0.13 nmoles/mg protein). Cisplatin treated rats showed a decrease in superoxide dismutase, catalase, glutathione peroxidase (GSH peroxidase), and glutathione reductase (GSH-reductase) activities (57%, 83%, 78% and 58% of control). Cochlear superoxide dismutase, catalase and GSH-reductase activities and MDA levels were restored in the rats injected with cisplatin plus MTBA, compared to cisplatin alone. It is concluded that the protection conferred by MTBA against cisplatin ototoxicity is associated with sparing of the cochlear antioxidant system. PMID- 9353847 TI - Antimuscarinic potency and bladder selectivity of PNU-200577, a major metabolite of tolterodine. AB - PNU-200577 (labcode DD 01 [(R)-N, N-diisopropyl-3-(2-hydroxy-5 hydroxymethylphenyl)-3-phenylpropanamine ) is a major pharmacologically active metabolite of tolterodine, a new muscarinic receptor antagonist intended for the treatment of an overactive bladder. In vitro, PNU-200577 produced a competitive and concentration-dependent inhibition of carbachol-induced contraction of guinea pig isolated urinary bladder strips (KB = 0.84 nM; pA2 = 9.14). In vivo, PNU 200577 was significantly more potent at inhibiting acetylcholine-induced urinary bladder contraction than electrically induced salivation in the anaesthetised cat (ID50 15 and 40 nmol.kg-1, respectively; P < 0.01). In radioligand binding studies carried out in homogenates of guinea-pig tissues and Chinese hamster ovary cell lines expressing human muscarinic m1-m5 receptors, PNU-200577 was not selective for any muscarinic receptor subtype. Thus, PNU-200577 is similar to tolterodine in terms of antimuscarinic potency, functional selectivity for the urinary bladder in vivo and absence of selectivity for muscarinic receptor subtypes in vitro. The results of this study clearly indicate that PNU-200577 contributes to the therapeutic action of tolterodine, in view of its high antimuscarinic potency, similar serum concentration and lower degree of protein binding. PMID- 9353849 TI - Arterial function in mineralocorticoid-NaCl hypertension: influence of angiotensin-converting enzyme inhibition. AB - Angiotensin-converting enzyme inhibitors have been suggested to improve the function of arterial endothelium and smooth muscle not only through inhibition of angiotensin II formation and reduction of blood pressure, but also via additional pathways, e.g. potentiation of endogenous kinins and enhancement of endothelial autacoid formation. Therefore, we investigated whether 10-week-long quinapril therapy (10 mg kg-1 day-1) could beneficially influence the function of mesenteric arterial rings in vitro in deoxycorticosterone-NaCl-treated Wistar Kyoto rats, a model of hypertension which is known to be resistant to angiotensin converting enzyme inhibition. The quinapril treatment had no long-term blood pressure-lowering effect nor did it reduce the associated cardiac hypertrophy in deoxycorticosterone-NaCl hypertension. In noradrenaline-precontracted arterial rings the endothelium-dependent relaxations to acetylcholine and adenosine 5' diphosphate as well as the endothelium-independent relaxations to nitroprusside and isoprenaline were clearly attenuated in the deoxycorticosterone-NaCl-treated rats. However, the quinapril therapy was without significant effect on any of these dilatory responses. In the presence of the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester, the relaxations to acetylcholine in untreated and quinapril-treated hypertensive animals were practically absent, whereas in normotensive rats distinct relaxations to higher concentrations of acetylcholine were still present. Interestingly, when endothelium-dependent hyperpolarization was prevented by precontracting the preparations with potassium chloride, no differences were found in relaxations to acetylcholine and adenosine 5' diphosphate between the study groups. Exogenous bradykinin induced small comparable contractions in endothelium-intact mesenteric arterial rings from all study groups. In conclusion, the 10-week-long quinapril therapy did not have any significant effects on arterial function in deoxycorticosterone-NaCl hypertensive rats. Therefore, the present results stress the roles of reduced blood pressure and diminished angiotensin II formation in the beneficial vascular effects of long-term angiotensin-converting enzyme inhibition in the present model of hypertension. Furthermore, since the relaxations to acetylcholine and adenosine 5'-diphosphate in the deoxycorticosterone-NaCl-treated rats were attenuated in the absence and presence of nitric oxide synthase inhibition but not under conditions which prevented hyperpolarization, impaired endothelium-dependent relaxation to agonists can be attributed to diminished endothelium-dependent hyperpolarization in this model of hypertension. PMID- 9353851 TI - Diabetes-induced protection from cisplatin nephrotoxicity is associated with impairment of energy-dependent uptake by renal cortex slices. PMID- 9353850 TI - Arterial responses to bradykinin after ramipril therapy in experimental hypertension. AB - Angiotensin-converting enzyme inhibitors have been shown to potentiate relaxations to kinins in several arteries, but the effects of long-term therapy on the responses to bradykinin in normotensive and hypertensive animals remain largely unknown. Therefore, the effects of 12-week-long ramipril therapy (1 mg kg 1 day-1) on responses of mesenteric arterial rings in vitro were studied in spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. Endothelium dependent relaxations of noradrenaline-precontracted rings to acetylcholine were similar in normotensive rats and ramipril-treated hypertensive rats and more pronounced than in untreated hypertensive group. Higher concentrations of bradykinin (0.1-1 microM) induced slight contractions in noradrenaline precontracted endothelium-intact rings of normotensive groups and untreated hypertensive group, whereas no response or a transient relaxation were observed in ramipril-treated hypertensive rats. Interestingly, in ramipril-treated hypertensive rats but not in the other groups, 20-min. pretreatment of arterial rings with ramiprilat unmasked or potentiated the relaxations to bradykinin, and these bradykinin-induced relaxations were effectively inhibited by the B2-kinin receptor antagonist Hoe-140. In conclusion, ramipril treatment clearly improved endothelium-dependent arterial relaxation to acetylcholine, and potentiated of even unmasked the dilatory response mediated via the endothelial B2-kinin receptor in spontaneously hypertensive rats. Since these enhancing effects on arterial relaxation in vitro could not be attributed to reduced breakdown of bradykinin, the present results suggest that long-term angiotensin-converting enzyme inhibition potentiated the actions of kinins at level of B2-kinin receptors. PMID- 9353853 TI - Conceptualization of schizophrenia: the symptom-oriented approach. AB - E. Bleuler proposed to establish the diagnosis of schizophrenia on the basis of fundamental symptoms which he presumed to be particularly linked with the primary deficiency. K. Schneider, on the other hand, based his diagnosis on the presence of symptoms recognizable without difficulty. Modern classifications include both Bleuler's fundamental and Schneider's first-rank symptoms in their diagnostic criteria. This may lead to erroneous attributions in view of the possibility that most first-rank and some of the symptoms Bleuler suspected to be fundamental may be unspecific reactions to different basic disturbances. It can therefore be expected that research on symptoms is better suited for aetiopathogenetic investigations than research based on diagnoses. Strategies to select those symptoms on which research should focus are described and recent findings of symptom-oriented studies are discussed. PMID- 9353854 TI - The deficit syndrome of schizophrenia: towards heterogeneity. AB - Since the turn of the century, psychiatrists have been concerned with both the unity and diversity of schizophrenia. From these early descriptions until now, many authors have attempted to delineate clinically meaningful subtypes within this disorder. In this connection, negative/positive subtyping has generated great interest. Carpenter and his team have emphasized the origin of the negative symptoms observed. They have proposed that primary enduring negative symptoms should be distinguished from transient negative symptoms resulting from treatment, depression or social deprivation and should be termed deficit symptoms. The validity of this subtyping is supported by clinical, biochemical or electrophysiological studies showing differences between deficit and nondeficit patients. PMID- 9353855 TI - Empirical assessment of the factorial structure of clinical symptoms in schizophrenia. A multisite, multimodel evaluation of the factorial structure of the Positive and Negative Syndrome Scale. The PANSS Study Group. AB - The Positive and Negative Syndrome Scale (PANSS) is widely used as a method for the assessment of symptoms of schizophrenia but the most complete model of how symptoms are structured has not been determined. Using the methods of confirmatory factor analysis with a large sample of 1,233 of schizophrenic subjects this study examined the goodness of fit of 20 previously proposed models. None of these proposed models met criteria for adequate fit to the empirical data. The sample was then stratified and half of the data was used to calibrate a new model. The model was validated in the second half of the data. The new pentagonal model uses 25 of the 30 items of the PANSS in 5 factors: positive, negative, dysphoric mood, activation, and and autistic preoccupation. Patients who varied widely in age, severity, and chronicity of illness did not differ in their overall symptom structure. The results of this study also implicated some problems in the validity of the PANSS as currently configured when used to assess symptoms of schizophrenia. PMID- 9353856 TI - Clinical heterogeneity of schizophrenia. AB - The heterogeneity of schizophrenia has led to a multitude of diagnostic criteria systems. Thus, the best strategy for schizophrenia research might be the use of several diagnostic systems simultaneously. This polydiagnostic approach can be associated with isolating subtypes of symptoms or patients. In this way, the authors present several approaches such as, first, dimensional approaches, second, cluster analyses, and third the selection of a very homogeneous subtype with standardized criteria. One homogeneous subtype can be represented by deficit schizophrenia according to Carpenter as defined by the Schedule of Deficit Syndrome. PMID- 9353857 TI - Family interaction and the course of schizophrenic illness. Results of a multivariate prospective study. AB - The aim of the present study was to assess the prognostic relevance of relatives' interactive behaviour towards the patient, as covered by the Munster Family Interview (MFI), to the further course of the schizophrenic illness. The MFI is a family interview (of the whole family, including the patient) designed to record the emotional family atmosphere based on the concept of expressed emotion (EE). The ratings take place directly after the interview on five scales (criticism, hostility, overinvolvement, resignation and warmth), the resignation scale being added to the 'classic' EE scales. Ninety-nine families of outpatients diagnosed with schizophrenia according to the DSM-III were examined with the MFI during a home visit. The patients were seen 1 and 2 years after the first examination. The target criteria selected for the prognostic significance of the interaction measurements were: rehospitalisation within 2 years; extent of symptoms after 1 year, and psychosocial skills after 1 year. The significance of the interaction dimensions was verified in regression models. The control variable used in the regression models was the Strauss-Carpenter scale. Regression models were produced for the total group and for a subgroup of moderately ill patients. All target criteria yielded serviceable prediction models. The most important variable for prediction was the control variable, the Strauss-Carpenter scale. However the interaction variables made additional contributions to the prognosis, especially in the subgroup of moderately ill patients. The best MFI scale for all the outcome criteria was resignation; criticism predicted only the symptomatology, and emotional overinvolvement the level of social functioning after 1 year. In conclusion, practical work with families of schizophrenic patients should emphasise the protective function of relatives towards patients more strongly. PMID- 9353858 TI - Numerical taxonomy applied to Leonhard's classification of endogenous psychoses. AB - Among the existing classifications in the field of psychiatry, Leonhard's comprises 35 forms of endogenous psychoses, allowing a more precise differentiation. Computerassisted mathematical methods of numerical taxonomy, frequently used in other fields of biological sciences, were used to check its coherence, definitely proving its taxonomic validity. Numerical values were assigned to signs and symptoms described by Leonhard, taking into account their frequency and diagnostical importance. The resulting data matrix was processed, obtaining the dissimilitude coefficients and the main components which validate Leonhard classification. This method can be applied to other psychiatric classifications validating them or displaying their weaknesses. PMID- 9353859 TI - On the notion of psychosis: the DSM-IV in perspective. AB - Using a historical overview and a conceptual analysis of the notion of psychosis, the DSM-IV suggestion to narrow down the notion of psychosis to delusions and hallucinations is examined. It is argued that the suggestion continues a trend in modern psychiatry, and that, perhaps more than others, it might lead to the (mis)identification of the notion of psychosis with the notion of error, which fails to demarcate disordered reality representation--as in delusions and hallucinations--from normal reality representation. Finally, the notion of (impaired) reality testing is presented as permitting such a demarcation, and it is concluded that it is therefore clinically useful and should be preserved, at least for the time being, in the notion of psychosis, including the narrow sense of the notion of psychosis as suggested in the DSM-IV. PMID- 9353860 TI - Non-invasive monitoring of bronchial inflammation in asthma. AB - The present consensus on the diagnosis and treatment of asthma relies on symptoms and lung function measurements for the monitoring of disease severity. Even though this probably remains the cornerstone of asthma management, the rapidly increasing insight into the pathogenesis and pathophysiology of the disease is presently leading to the development of more direct measurements of airway inflammation, which may provide potentially relevant information on its clinical course and prognosis. However, at present none of these has sufficiently been validated for current use in monitoring patients with asthma. First, there are new ways of looking at symptoms and lung function. Careful measurements of symptoms by visual analogue scale (VAS) are suggesting that inflammatory activity within the airways can be subjectively perceived, a sensitivity which may be blunted in patients with brittle asthma. In addition, modern physiological parameters, such as the degree of bronchodilatation following a deep breath (M/P ratio), are strongly associated with airway inflammation. Second, there are multiple cellular and/or soluble markers of inflammation in peripheral blood (using PCR, in situ hybridization, flow cytometry, or circulating mediators and cytokines) and in urine (LTE4, EPX). Recently this has been extended by similar measurements in hypertonic saline-induced sputum (cell differentials and specific stainings on cytospins, flow cytometry, and levels of e.g. ECP, IL-5, IL-8). Finally, mediators and cytokines in the condensate of exhaled air (H2O2, leukotrienes, IL-5?) as well as exhaled NO are currently under evaluation. Adding such markers of airway inflammation as guides in asthma therapy is potentially useful. As a first step towards such a new approach we have recently shown that adding the reduction of airway hyperresponsiveness to the aims of asthma therapy leads to a better clinical as well as histological outcome after two years of treatment. In conclusion, there are new and exciting perspectives in the monitoring of disease severity in asthma in the future. Longitudinal studies presently ongoing will elucidate which parameter is potentially most useful in guiding asthma management. PMID- 9353861 TI - Lung diseases in pregnancy. AB - When physicians encounter pregnant patients with respiratory complaints, they face a challenging set of clinical problems. To understand the clinical cardiopulmonary manifestations of diseases occurring during pregnancy, knowledge of the basic physiologic changes during pregnancy is necessary. The most prevalent chest related complaint in pregnant women is shortness of breath, which is in most cases due to an unpleasant awareness of physiological gestational hyperventilation. Lung diseases which are frequently seen in young people, such as bronchial asthma, occur with comparable prevalence in pregnant women. Clinical symptoms, diagnosis and treatment of most diseases do not differ from those in the nonpregnant state. However, pharmacotherapy presents unique aspects, since not only may pharmacokinetics differ, but the fetus must also be assumed to be a recipient of the drug. PMID- 9353862 TI - Circulatory abnormalities in cirrhosis with focus on neurohumoral aspects. AB - Patients with cirrhosis exhibit characteristic hemodynamic changes with a hyperkinetic circulation and an abnormal distribution of the blood volume and neurohumoral regulation. Their plasma and noncentral blood volumes are increased, and the central and arterial blood volume and systemic vascular resistance are decreased. A peripheral arterial vasodilatation may be of pathogenic importance to the low systemic vascular resistance as it directly correlates to the degree of central hypovolemia. It may therefore be an important element in the development of the low arterial blood pressure and hyperkinetic circulation in cirrhosis. Various vasodilators such as atrial natriurectic peptide, calcitonin gene-related peptide, adrenomedullin, and nitric oxide are among potential candidates in the arterial vasodilatation in cirrhosis. Besides enhanced sympathetic nervous activity, activation of the renin-angiotensin-aldosterone system, and elevated circulating vasopressin, endothelin-1 may also be implicated in the hemodynamic counter-regulation in cirrhosis. Recent research has focused on the assertion that the hemodynamic and neurohumoral abnormalities in cirrhosis are part of a general circulatory dysfunction influencing the course of the disease. PMID- 9353863 TI - Atrial natriuretic peptide: renal effects in cirrhosis of the liver. AB - Atrial natriuretic peptide is one of a family of natriuretic peptides thought to play a role in the altered sodium balance of advanced liver disease and ascites. Its level is usually increased in the plasma of cirrhotic patients, probably due to relative plasma volume expansion. When exogenous ANP is administered intravenously to dogs or rats with experimental liver cirrhosis and ascites, an heterogeneous natriuretic response is obtained with about half of the population not responding. Similar observations are recorded for patients with clinical cirrhosis. In dogs, attenuation of the ANP-induced natriuresis may depend on a reduction in renal cortical bradykinin activity. In patients with cirrhosis, the ability to release ANP in response to central volume expansion is dissociated from the accompanying natriuresis. Attenuation of the renal tubular response to ANP in this setting may be correlated to the degree of intrahepatic sinusoidal hypertension and associated augmented reflex sympathetic nervous activity to the kidneys. Actual tubular resistance to ANP may be due to reduced Na+ delivery to the inner medullary collecting duct and/or increased degradation of cyclic guanosine monophosphate. PMID- 9353864 TI - Arachidonic acid derivatives and renal function in liver cirrhosis. AB - Prostaglandins (PGs) are arachidonic acid (AA) derivatives via the PG endoperoxyde H synthase (PGHS) complex. Two PGHS isoforms have been recognized, constitutive (PGHS-1) and inducible (PGHS-2), respectively. Within the kidney, vascular endothelium mainly produces PGI2; the whole glomerulus synthesizes several prostanoids, the predominant AA metabolite in humans being PGI2; tubules and medullary interstitial cells produce mainly PGE2. Renal PGs modulates the action of other hormones and autacoids involved in the regulation of renal hemodynamics, glomerular filtration and the renal handling of sodium and water. Renal PGs are, at least in part, excreted into urine. Measurement of urinary PGs or their metabolites has been found to provide a reliable estimation of basal as well as stimulated PG synthesis. Patients with cirrhosis of the liver show an increased renal synthesis of vasodilating PGs, as indicated by the high urinary excretion of PGs and/or their metabolites. Administration of nonsteroidal anti inflammatory drugs (NSAIDs) to these patients causes a profound reduction in renal blood flow and glomerular filtration rate, a reduction in sodium excretion, and an impairment of free water clearance. These data clearly indicate that the increased renal synthesis of vasodilating PGs has a relevant role in maintaining renal hemodynamics, sodium and water excretion in a clinical setting characterized by a reduction of effective plasma volume and a striking activation of the major vasoconstricting systems, namely the renin-angiotensin-aldosterone, the sympathetic nervous system, and vasopressin. Patients with hepato-renal syndrome have a reduced renal synthesis of vasodilating PGE2 in the setting of a striking activation of endogenous vasoconstrictors and a maintained or increased renal production of thromboxane A2. Therefore, an imbalance between vasoconstricting systems and the renal vasodilator PGE2 was proposed to explain the renal failure observed in this condition. The urinary excretion of 2-3-dinor 6-keto-PGF1 alpha, an index of systemic PGI2 synthesis, is increased in patients with cirrhosis and hyperdynamic circulation, thus raising the possibility that systemic synthesis of PGI2 may contribute to the arterial vasodilatation of these patients. Finally, administration of exogenous prostanoids to patients with cirrhosis is not effective either in ameliorating renal function or in preventing the deleterious effect of NSAIDs. PMID- 9353865 TI - Bile acids, oxidative stress, and renal function in biliary obstruction. AB - Renal dysfunction occurs in patients with biliary obstruction. Plasma accumulation of bile acids and oxidative stress have been proposed as contributory factors. Bile acids can alter the renal handling of electrolytes and water by blocking the Na(+)-H+ antiport in the tubule. Oxidative stress, defined as an imbalance between radical generating systems and radical scavenging systems giving rise to free radical induced tissue damage, occurs in patients with liver disease. Bile acids cause oxidative damage to tubular cell membranes by stimulating the generation of oxygen free radicals from mitochondria, as well as promoting their release from neutrophils and macrophages. Oxidative stress can promote the formation of a variety of vasoactive mediators including endothelin 1, cysteinyl leukotrienes, as well as the F2-isoprostanes, endogenous products of lipid peroxidation. These mediators can each affect renal function directly by causing renal vasoconstriction or decreasing the glomerular capillary ultrafiltration coefficient, and thus reduce glomerular filtration rate. Collectively, these factors contribute to the onset of renal failure in patients with biliary obstruction. PMID- 9353866 TI - Hepatorenal syndrome: emerging perspectives. AB - Progressive oliguric renal failure (designated hepatorenal syndrome) commonly complicates the course of patients with advanced hepatic disease. Despite the severe derangement of renal function and ominous prognosis when renal failure develops, minimal and inconsistent pathologic abnormalities of the kidneys are found at autopsy. Furthermore, the kidneys, if transplanted, are capable of normal function, which supports the concept that the renal failure is functional and potentially reversible. In contrast to patients with classical acute renal failure (ATN), hepatorenal syndrome patients manifest characteristic alterations of renal function including (1) relatively hyperosmolar urine; (2) high creatinine urine to plasma (U:P) ratio, and (3) a very low urine sodium concentration (< 10 mEq/L). During the past several years we have witnessed new insights into both the pathophysiology and the therapeutics of this syndrome. The application of new methodology such as tracer kinetics has more rigorously delineated the role of a number of pathogenic mechanisms, including activation of the sympathetic nervous system. The characterization of endothelin and nitric oxide-arginine pathway and their roles in biology and medicine has provided additional new insights with regard to the pathogenesis of hepatorenal syndrome. For example, nitric oxide has been proposed to constitute a mediator of both the hyperdynamic circulation and renal failure. Finally, recently initiated therapeutic approaches lend a note of optimism to the future management of a syndrome that is so often incompatible with recovery. These include the acceptance of orthotopic liver transplantation as definitive treatment for patients with end-stage liver disease and attempts to improve renal function by use of transjugular intrahepatic porto-systemic shunt. Hopefully, ongoing and future clinical trials will establish the precise contribution of each of these treatment modalities and their respective roles in the therapeutic armamentarium. PMID- 9353867 TI - Extracorporeal blood purification in the management of patients with hepatic failure. AB - With the increasing success of orthotopic liver transplantation, the time has come for a reassessment of the role of extracorporeal blood purification in the management of patients with liver failure. In those patients with combined liver and kidney failure, both standard hemodialysis and the newer continuous renal replacement therapies have been found to be helpful in maintaining fluid, electrolyte, and acid base balance. In those patients with liver failure but without kidney dysfunction, extracorporeal purification techniques have been found useful for the removal of hepatic toxins and in facilitating the replacement of clotting factors. This review will outline the rationale and documented utility of a wide variety of blood purification modalities used in the management of patients with liver failure. PMID- 9353868 TI - Selected lines of Aedes aegypti with persistently distorted sex ratios. AB - A breeding scheme to isolate X chromosomes sensitive to drive by the T8 (Trinidad) Y chromosome of Aedes aegypti (the MD haplotype) is reported. Crosses with an Australian strain Th.I (Thursday Island) revealed not only sensitive and resistant X chromosomes but also some with the capacity to drive against the T8 Y chromosome. Four strains were created in which sex ratio was male-distorted (28 36 per cent Female) for 10 generations, with no regression towards sexual parity. The proportion of females varied significantly between generations in each of the four strains. Further selection produced strains with normal sex ratios, capable of generating fewer than 15 per cent Female on outcrossing to T8 males. PMID- 9353869 TI - Recent case histories of food product-metal container interactions using scanning electron microscopy-x-ray microanalysis. AB - Scanning electron microscopy (SEM) and x-ray microanalysis (EDS) were used to investigate stress corrosion cracking (SCC) in plain tinplate food cans, product discoloration, filiform corrosion, pitting/perforation corrosion, and loss of coating adhesion in enameled food cans. Intergranular SCC of uncoated tinplate occurred in canned pineapple juice. This is a rare occurrence in acid food products. The fracture developed in the can headspace only at the product line/metal interface by an interaction of the detinned metal surface with stress inducing compounds in the product. Black discoloration of rice granules in the can headspace of a chicken and rice product was caused by the formation of metal sulfides. The source of the metal contamination was traced to metallic dirt in the coating on the tin-free-steel end. Blackening of light tan-colored olives and brine occurred after 1 year of storage in enameled tinplate containers as a result of pitting corrosion at the side seam weld. The cause of the pitting was a defective side seam stripe which failed to protect the weld. A perforation problem occurred at the score line of aluminum-tinplate bimetallic cans. The failure was caused by the high chloride content of the fruit product. Filiform corrosion resulted in perforations that occurred on the outside surfaces of two piece tin-free steel cans packed with tuna. The cause of the corrosion was related to scratch defects in the exterior coating and the presence of chloride and sulfate cannery residues in the corroded areas. An enamel adhesion failure developed inside two-piece tin-free steel cans that contained mushrooms. Wrinkles in the coating which were introduced during can manufacture were cracked. The fractures were pathways for product-steel interaction which resulted in container failure. PMID- 9353870 TI - Reduction of vanadate to vanadyl by a strain of Saccharomyces cerevisiae. AB - Three strains of Saccharomyces cerevisiae, SC-1, DBVPG 6173 and DBVPG 6037, were studied for vanadate resistance in complex Sabouraud medium since they did not thrive in different minimal media (yeast nitrogen base with and without amino acids). The strain SC-1 was resistant up to 16 mM of vanadate, whereas the strains DBVPG 6173 and DBVPG 6037 were inhibited by 8 mM and 4 mM vanadate, respectively. The vanadate resistance in strain SC-1 was constitutive and due to the reduction of this oxyanion to vanadyl, which was detected by EPR spectroscopy and visible spectroscopy. The transformation of vanadate to vanadyl took place during the exponential growth phase; 10 mM of vanadate was reduced to vanadyl outside the cells since the oxyanion was not detected in the cell biomass and only a negligible concentration of vanadyl (25 nmoles mg-1 cells dry weight) was found in the biomass. The other two vanadate-sensitive yeast strains only accumulated vanadate and did not reduce the oxyanion to vanadyl. PMID- 9353872 TI - Purification of a fully metal-depleted Cu, Zn superoxide dismutase from copper deficient rat liver. AB - A copper-deprived form of the enzyme Cu, Zn superoxide dismutase was identified in the liver of rats made copper-deficient by dietary restriction. In homogenates of such livers Cu, Zn superoxide dismutase presents a dis-homogeneous electrophoretic profile with respect to the native enzyme. When rat liver extracts were treated with exogenous copper an electrophoretic pattern resembling the native one was observed. Enzyme purified by chromatography on DE-52 resin shows two major components, one corresponding to genuine, native enzyme and another one, eluting at higher ionic strength. The latter protein (Fraction II) consists of several isoforms which show the same characteristics of the native superoxide dismutase as far as immunoreactivity and molecular weight are concerned, but with decreased contents of copper and zinc. Its catalytic constant, referring to copper content, was 15 times lower than that obtained for the native enzyme. Moreover, the catalytic power of purified Fraction II was not regained upon incubation with copper. The occurrence of a superoxide dismutase void of metals confirms the hypothesis that this protein plays a dual physiological role: in metal metabolism and in superoxide anion dismutation. PMID- 9353871 TI - Liver iron depletion and toxicity of the iron chelator deferiprone (L1, CP20) in the guinea pig. AB - The use of the iron chelator deferiprone (L1, CP20, 1,2-dimethyl-3-hydroxypyrid-4 one) for the treatment of diseases of iron overload and other disorders is problematic and requires further evaluation. In this study the efficacy, toxicity and mechanism of action of orally administered L1 were investigated in the guinea pig using the carbonyl iron model of iron overload. In an acute trial, depletion of liver non-heme iron in drug-treated guinea pigs (normal iron status) was maximal (approximately 50% of control) after a single oral dose of L1 of 200 mg kg-1, suggesting a limited chelatable pool in normal tissue. There was no apparent toxicity up to 600 mg kg-1. In each of two sub-acute trials, normal and iron-loaded animals were fed L1 (300 mg kg-1 day-1) or placebo for six days. Final mortalities were 12/20 (L1) and 0/20 (placebo). Symptoms included weakness, weight loss and eye discharge. Iron-loaded as well as normal guinea pigs were affected, indicating that at this drug level iron loading was not protective. In a chronic trial guinea pigs received L1 (50 mg kg-1 day-1) or placebo for six days per week over eight months. Liver non-heme iron was reduced in animals iron loaded prior to the trial. The increase in a wave latency (electroretinogram), the foci of hepatic, myocardial and musculo-skeletal necrosis, and the decrease in white blood cells in the drug--treated/normal diet group even at the low dose of 50 mg kg-1 day-1 suggests that L1 may be unsuitable for the treatment of diseases which do not involve Fe overload. However, the low level of pathology in animals treated with iron prior to the trial suggests that even a small degree of iron overload (two-fold after eight months) is protective at this drug level. We conclude that the relationship between drug dose and iron status is critical in avoiding toxicity and must be monitored rigorously as cellular iron is depleted. PMID- 9353873 TI - Effects of Pb2+, Ni2+, Hg2+ and Se4+ on cultured cells. Analysis of uptake, toxicity and influence on radiosensitivity. AB - Effects of Pb2+, Ni2+, Hg2+ and Se4+ on cultured human glioma U-343MG cells were investigated considering uptake, toxicity and, in combination with radiation, clonogenic cells survival. The cells were exposed to 0-100 microM of the metals for a week before the evaluation. The tests showed a tendency to toxicity with 10 microM nickel although not significant (P > 0.05). Selenium, lead and mercury exerted a significant toxicity (P < 0.05) at 2.5 microM, 10 microM and 1 microM, respectively. To challenge the clonogenic cell survival capacity, the cells were irradiated with 60Co photons after being exposed to the highest nontoxic concentration of the different metals. The clonogenic cell survival tests, after irradiation, showed no significant change if the cells were exposed to 5 microM nickel, 0.5 microM selenium or 5 microM lead compared with those not exposed. Mercury, 0.1 microM, gave a relative reduction in survival compared with only irradiated cells of 58 +/- 17%. Thus, only mercury affected the radiation-induced damage and/or repair. When exposed to the highest nontoxic concentrations of the different metals, the cultures did not display a significant uptake ratio (metal concentration ratio of exposed cells to control cells) of nickel (3.1 +/- 3.3), only a small uptake ratio of selenium (4.0 +/- 0.4), while there was a large uptake ratio of both lead (2.6 +/- 1.7) x 10(2) and mercury (1.5 +/- 0.2) x 10(1). The results indicated that nickel was neither especially toxic nor influenced the clonogenic cell survival after irradiation. Mercury was more toxic and also influenced the radiation sensitivity. Lead was taken up strongly but did not influence the radiation sensitivity. Selenium accumulated but gave no detectable effect on the radiation sensitivity. PMID- 9353875 TI - Sublethal effects of experimental exposure to mercury in European flat oyster Ostrea edulis: cell alterations and quantitative analysis of metal. AB - Oysters display a diversity of uptake mechanisms for metallic elements and distribution in the target organs, namely gills and the digestive gland. Various tissues of the flat oyster, Ostrea edulis, were studied following experimental exposure to 0.025 microM (5 micrograms l-1) of mercury, for up to 34 days. All animals survived the treatment. Data indicate Hg accumulation in gill tissue with a maximum concentration of 38.76 micrograms g-1 dry weight after 25 days of exposure. Hg levels were lower in remaining tissues, in which the maximum concentration (18.47 micrograms g-1 dry weight) was reached after 18 days of exposure. After these times, concentration in both tissues decreased. Results show that oysters can accumulate Hg from the environment, without their survival being affected during the experimental period. Structural alteration of epithelial tissues of gill and digestive gland of flat oyster was comparable with effects described for other metallic elements in bivalve molluscs. Interstitial tissue was disorganized in the digestive gland, and ultrastructural changes in intracellular endomembranes were detected in epithelial cells of the digestive gland after 18 days of treatment. After 25 days, absorptive epithelial cells of gills showed highly dilated, swollen microvilli. These intracellular alterations are parameters of the incipient response to the accumulation of mercury. PMID- 9353874 TI - Effect of nicotinic acid on zinc and iron metabolism. AB - Nicotinic acid has functional groups capable of forming complexes with trace metals. The present study examines the effect of nicotinic acid supplementation on absorption and utilization of zinc and iron. In vitro zinc uptake by human erythrocytes was studied using blood samples of 10 healthy subjects. It was found that 8 mumoles nicotinic acid or NADP increased 65Zn uptake by 38.9% and 43.1% in fasting, and by 70.9% and 28.1% in postprandial conditions. In animal experiments, nicotinic acid supplementation to finger millet based diet resulted in significant enhancement of percent zinc absorption, liver zinc and growth of weanling mice (P < 0.05). When mice were fed with nicotinic acid-deficient, adequate and -excess synthetic diets for four weeks it was observed that, in comparison with the nicotinic acid-deficient diet, percent zinc absorption, intestinal zinc, percent haeomoglobin and liver iron increased significantly under nicotinic acid-adequate and -excess conditions. The results obtained suggested that nicotinic acid, in addition to its known effect on growth and metabolism, may be playing an important role in enhancing zinc and iron utilization. PMID- 9353876 TI - Inorganic mercury binding to fish oocyte plasma membrane induces steroidogenesis and translatable messenger RNA synthesis. AB - Both in vitro and in vivo HgCl2 treatment demonstrated a remarkably high rate of progesterone synthesis accompanied by a low rate of conversion to 17 beta estradiol in the oocyte of Channa punctatus. On depuration, however, there was a reversal of the steroidogenic scenario with a low progesterone and high estradiol level. The accumulation of progesterone was positively correlated with the significant increase in 3 beta-hydroxysteroid dehydrogenase activity in the Hg treated fish. Thus, it was clear that at the early stage of intoxication Hg does play a role in the induction of 3 beta-hydroxysteroid dehydrogenase in the oocyte of fish at the spawning stage. The induction of this enzyme was found to be mediated by specific binding of Hg to the plasma membrane Na(+)-K(+)-ATPase (Bmax: 14 nmoles mg-1 protein; Ka 1.14 x 10(8) moles) and increase in the specific messenger RNA translating 3 beta-hydroxysteroid dehydrogenase. It is concluded that inorganic mercury is able to initiate translatable messenger RNA synthesis in fish oocyte at a low degree of intoxication. PMID- 9353877 TI - Binding of vanadate to human erythrocyte ghosts and subsequent events. AB - Spectroscopic techniques were used to investigate the interaction between vanadate and human erythrocyte ghosts. Direct evidence from 51 V nuclear magnetic resonance (NMR) studies suggested that the monomeric and polymeric vanadate species may bind to the anion binding sites of band 3 protein of the erythrocyte membrane. The results of 51V NMR studies and the quenching effect of vanadate on the intrinsic fluorescence of the membrane proteins indicated that in the low concentration range of vanadate (< 0.6 mM), monomeric vanadate binds mostly to the anion sites of band 3 protein with the dissociation constant close to 0.23 mM. The experiments of sulfhydryl content titration by the method of Ellman and residue sulfhydryl-labeled fluorescence spectroscopies clearly displayed that vanadate reacts directly with sulfhydryl groups. The appearance of the anisotropic election spin resonance (ESR) signal of vanadyl suggests that a small (c.3%) amount of vanadate was reduced by sulfhydryl groups of membrane proteins. The fluidity and order of intact ghost membrane were reduced by the reaction with vanadate, as shown by the ESR studies employing the protein- and lipid-specific spin labels. It was concluded that although vanadates mainly bind to band 3 protein, a minor part of vanadate may oxidize the residue sulfhydryl groups of membrane proteins, and thus decrease the fluidity of erythrocyte membrane. PMID- 9353878 TI - Selenium levels in infant milk formula. AB - Twenty-four brands of commercial infant milk formula were collected and analysed for selenium by inductively coupled plasma spectrometry with the hydride t-system after an acid digestion procedure. The mean selenium concentration was 49.0 +/- 11.55 micrograms l-1, with a range from 26-68 micrograms l-1, resulting in an adequate daily selenium intake for infants aged from zero to six months consuming 0.75 l milk daily as set by the US National Research Council in 1989. PMID- 9353879 TI - The pH dependent properties of metallotransferrins: a comparative study. AB - The dependence on pH of the absorption and circular dichroic spectra of iron(III), cobalt(III) and copper(II) transferrins has been (re)investigated. In the alkaline region, the CD profiles of iron(III) and cobalt(III) transferrin are essentially pH independent up to pH 11; only for very high pH values (pH > 11) is breakdown of the cobalt(III) and iron(III) transferrin derivatives observed, without evidence of conformational rearrangements. By contrast, the CD profiles of copper transferrin show drastic changes in shape around pH 10; these spectral changes, which are fitted to a pKa of approximately 10.4, are interpreted in terms of a substantial rearrangement of the local environment of the copper ions at high pH. Although the CD spectra of copper transferrin at alkaline pH strictly resemble those observed upon addition of modifier anions, the mechanism of site destabilization in the two cases is different; at variance with the case of modifier anions, our results suggest that the high pH form of copper transferrin still contains the synergistic anion. A 13C NMR experiment has confirmed this view. In the acidic region, iron(III) and cobalt(III) transferrins are stable down to pH approximately 6. For lower pH values progressive metal detachment is observed without evidence of conformational changes; around pH 4.5 most bound metals are released. In the case of the less stable copper-transferrin, metal removal from the specific binding sites is already complete around pH 6.0; in concomitance with release from the primary sites, binding of copper ions to secondary sites is observed. Additional information has been gained from CD experiments in the far UV. The pH dependent properties of iron(III), cobalt(III) and copper(II) transferrin are discussed in the frame of the present knowledge of transferrin chemistry, particular emphasis being attributed to the comparison between tripositive and bipositive metal derivatives. PMID- 9353880 TI - Urinary mercury levels in females: influence of skin-lightening creams and dental amalgam fillings. AB - The influence of application of skin-lightening creams and dental amalgam fillings on the urinary mercury (Hg) level was evaluated in 225 females (ages 17 to 58 years) living in Riyadh, capital of Saudi Arabia. The arithmetic mean of the urinary Hg level was 6.96 +/- 20.43 micrograms 1(-1), in the range 0 to 204.8 micrograms 1(-1). The mean urinary Hg level adjusted by creatinine (Cr) was 11.22 +/- 37.23 micrograms g-1 Cr, in the range 0 to 459.37 micrograms g-1. No significant difference in urinary Hg was noted between the females regarding the use of skin-lightening creams. On the other hand, results showed that urinary Hg concentration was influenced by the use and number of dental amalgam fillings. No women were identified with symptoms or signs that could be attributed to Hg intoxication. Urine analyses for creatinine, urea, uric acid, phosphorus, magnesium, glucose and calcium showed significant correlation with urinary Hg. This suggests that chronic exposure to Hg may be associated with a deterioration of renal function. PMID- 9353881 TI - The protective effect of zinc sulphate pretreatment against duodenal ulcers in the rat. AB - Exogenously administered zinc compounds have been shown to possess antiulcer activity in the development of gastric lesions. The aim of this study was to investigate the effects of zinc sulphate pretreatment of rats on cysteamine induced duodenal ulcers and to correlate them with changes in zinc serum and tissue levels. Atomic absorption spectrophotometry was used to determine zinc serum and tissue concentrations in all animal groups. Cysteamine produced marked duodenal ulceration in control animals 24 h after application, with an increase in endogenous zinc tissue concentrations and a marked decrease in serum concentrations. Zinc sulphate (20, 40 or 80 mg kg-1) applied per os one hour prior to cysteamine application inhibited the development of duodenal lesions in a dose-related manner. The application of zinc sulphate in a single intraperitoneal (i.p.) application (80 mg kg-1) did not, however, prevent the formation of duodenal lesions. In order to assess zinc absorption from the gastrointestinal tract, one group of rats received a single oral dose of zinc sulphate (80 mg kg-1) without cysteamine application. The observations of this study seem to indicate that zinc plays an important cytoprotective role in duodenal ulcer disease. PMID- 9353882 TI - Lead effects on structural and functional cellular parameters in human red cells from a prenatal hematopoiesis stage. AB - An attenuation (or reversion) of the prolytic effect of lead on neonatal red cells is observed in iso- or hypotonic low ionic strength media. This effect correlates neither with concomitant activation of K+ (Ca2+ or Pb2+) channels nor with volume reduction. Neonatal erythrocytes were used in this study owing to their greater cellular density, as compared with adult red cells, for the above mentioned channels. The attenuation-reversion effect would be mediated through lead interactions with the cytoskeleton, a structure that is the limiting factor for red cell lysis in low ionic strength media. PMID- 9353883 TI - Influence of zinc-saccharide complexes on some haematological parameters in rats. PMID- 9353884 TI - Uptake of Ga-67 into rat cerebral hemisphere and cerebellum. Comparison with Fe 59. AB - Transferrin and transferrin receptors play an important role in the transport of iron into the brain. To determine whether gallium enters the brain by the same mechanism, uptakes of 67Ga and 59Fe have been compared under controlled conditions. Rates of gallium penetration into brain (K(in)) were four times slower than those for 59Fe. K(in) for 67Ga when infused with citrate were 0.88 +/ 0.24 and 0.94 +/- 0.39 x 10(-3) ml g-1h-1 for cerebral hemisphere and cerebellum, respectively. When infused as the transferrin complex, 67Ga uptake into the brain was not different from that when infused with citrate. The presence of the anti-transferrin receptor antibody OX-26 significantly reduced uptake of 59Fe by 60% and 64% into cerebral hemisphere and cerebellum, respectively. By contrast, pretreatment of rats with OX-26 enhanced the uptake of 67Ga into brain, particularly when infused with citrate; mean increases in uptake of 67Ga were 120% and 144% for cerebral hemisphere and cerebellum, respectively. Purified 67Ga-transferrin was also taken up into both brain regions examined in the presence of OX-26. These results indicate that the transport of non transferrin bound gallium is an important mechanism for gallium uptake into brain. PMID- 9353885 TI - Analysis of magnetic material in the human heart, spleen and liver. AB - Isothermal remanent magnetization (IRM) acquisition and alternating field (A.F.) demagnetization analyses were performed on human heart, spleen and liver samples resected from cadavers. The magnetic properties of the samples were measured both at 77K and at 273K. A.F. demagnetization was performed at 273K. Results from the analyses of the tissue indicate the presence of ferromagnetic, fine-grained, magnetically interacting particles which, due primarily to magnetic properties, are thought to be magnetite and/or maghemite. The presence of superparamagnetic particles can be inferred from the increase in saturation IRM values when measured at 77K compared with measurements at 273K and the decay of remanent magnetization upon warming from 77K. The concentration of magnetic material (assuming it is magnetite or maghemite) in the samples varies from 13.7 ng g-1 to 343 ng g-1, with the heart tissue generally having the highest concentration. The presence of magnetic material in these organs may have implications for the function of biogenic magnetite in the human body. PMID- 9353886 TI - Urinary excretion of mercury, copper and zinc in subjects exposed to mercury vapour. AB - The excretion of mercury, copper and zinc in urine, and mercury in whole blood and plasma, was determined in 40 chloralkali workers exposed to mercury vapour and 40 age-matched referents. The Hg concentrations in whole blood, plasma and urine were higher in the exposed group (35 nmol l-1, 30 nmol l-1, and 11.5 nmol mmol-1 creatinine, respectively) in comparison with the reference group (15 nmol l-1, 6.3 nmol l-1, and 1.8 nmol mmol-1 creatinine, respectively). The urinary copper excretion was similar in the two groups, while U-Zn excretion was significantly higher (P = 0.04) in the exposed group, median 0.83 mumol mmol-1 creatinine versus 0.76 munmol mmol-1 creatinine in the reference group. In a subgroup of exposed workers with current U-Hg above 11.5 nmol l-1 mmol-1 creatinine (20 micrograms g-1 creatinine) the median U-Zn was 1.1 mumol mmol-1 creatinine. In both groups smokers had high U-Zn levels than non smokers. When both U-Hg and smoking were taken into account in a linear regression model, there was a significant association between U-Hg and U-Zn in the combined group of exposed and referents (P = 0.002). This study indicates that mercury exposure in humans, as in animals, causes increased urinary excretion of zinc. The mechanisms may be induced synthesis of metallothionein in the kidneys, displacement of Zn from preexisting metallothionein by Hg, or a decreased reabsorption of zinc in the kidneys owing to a slight tubular dysfunction. PMID- 9353887 TI - Aluminum site structure in serum transferrin and lactoferrin revealed by synchrotron radiation X-ray spectroscopy. AB - The Al site structure of serum transferrin and lactoferrin is investigated using X-ray absorption near edge structure (XANES) spectroscopy. Al K-edge spectra in the mono- and dialuminum forms of the proteins have been recorded for the first time. Our results show that the aluminium ion is hexa-coordinated in an octahedral-like symmetry and that the monoaluminum form, where only the C terminal binding site is saturated, has an increased structural distortion around the metal site. PMID- 9353888 TI - Degree of DNA unwinding caused by the binding of aclacinomycin A. AB - The effect of drug binding on the geometry of DNA duplex (plasmid pBR322) has been examined using topoisomerase I relaxation followed by gel electrophoresis. The binding of one molecule of aclacinomycin A was found to cause an unwinding of the DNA double helix by an angle of 8 +/- 2 degrees in aqueous solution at 37 degrees C. The unwinding angle of daunomycin was 12 +/- 2 degrees, and that of ethidium bromide 15 +/- 3 degrees. To determine the unwinding angle, precise determination of the equilibrium constant of drug-DNA binding-dissociation reaction in the same buffer as that for the topoisomerase reaction (at 37 degrees C) was indispensable. This determination was made by ultraviolet absorption measurement of the same plasmid-drug system, followed by a Scatchard plot and analysis using McGhee-von Hippel's excluded site model. For the aclacinomycin pBR322 system, the binding constant (K) was 7.2 x 10(4) M-1, and the number of base pairs in the single site of drug binding (n) was 4.0. For daunomycin-pBR322, K = 7.1 x 10(4) M-1 and n = 3.4, and for ethidium-pBR322, K = 4.0 x 10(4) M-1 and n = 3.3. On the basis of these experimental results, the possible role of the sugar moieties of these antitumour drugs, as well as that of intercalating chromophores, was discussed. PMID- 9353889 TI - Indonesian medicinal plants. XXI. Inhibitors of Na+/H+ exchanger from the bark of Erythrina variegata and the roots of Maclura cochinchinensis. AB - Through bioassay-guided separation of the methanol extracts of Indonesian medicinal plants, three inhibitors of the Na+/H+ exchange system, erythrinin B (2), euchrenone b10 (3), and 1,3,5-trihydroxy-4-(3-methylbut-2-enyl)xanthen-9-one (4), were isolated from the bark of Erythrina variegata (Fabaceae) (for 2) and the roots of Maclura cochinchinensis (Moraceae) (for 2, 3, 4). Compounds 2, 3, and 4 significantly inhibited the Na+/H+ exchange system of arterial smooth muscle cells, with minimum inhibitory concentrations of 1.25, 1.25, and 10 micrograms/ml, respectively. Three new prenylated xanthones named isocudraniaxanthones B (5) and A (7) and isoalvaxanthone (9) were also isolated from M. cochinchinensis and the chemical structures were elucidated on the bases of their chemical and physicochemical properties. PMID- 9353890 TI - Synthetic studies of vitamin D analogs. XXIV. Synthesis of active vitamin D3 analogs substituted at the 2 beta-position and their preventive effects on bone mineral loss in ovariectomized rats. AB - Analogs related to 1 alpha,25-dihydroxy-2 beta-(3-hydroxypropoxy)vitamin D3 (ED 71) (4), oxa-type and carba-type analogs of vitamin D3 bearing substituents at the 2 beta-position of 1 alpha,25-dihydroxyvitamin D3 (1), were synthesized from the alpha-epoxides (6 and 13). Three analogs, ED-71 (4) and two carba-type analogs (16 and 26), showed potent preventive effects on bone mineral loss in pre osteoporosis model rats. ED-71 (4) was concluded to be an optimized analog and a promising candidate for the treatment of osteoporosis. PMID- 9353891 TI - Novel renin inhibitors containing (2S,3S,5S)-2-amino-1-cyclohexyl-6-methyl-3,5 heptanediol fragment as a transition-state mimic at the P1-P1' cleavage site. AB - A series of renin inhibitors containing the (2S,3S,5S)-2-amino-1-cyclohexyl-6 methyl-3,5-heptanediol (2-amino-3,5-anti-diol) fragment as a novel transition state mimic was synthesized, and their biological activities were evaluated. All of the synthesized compounds containing the 2-amino-3,5-anti-diol fragment at the P1-P1' position showed high in vitro renin-inhibitory activity with IC50 values in the 10(-8)-10(-10) M range, and most of them caused a reduction of blood pressure when administered orally to salt-depleted, conscious marmosets. The inhibitor (29) with the 4-hydroxypiperidine residue at the P4 position showed the highest activity in terms of both potency and duration of the blood pressure lowering effect. PMID- 9353892 TI - Potent NK1 receptor antagonists: synthesis and antagonistic activity of various heterocycles with an N-[3,5-bis(trifluoromethyl)benzyl]-N-methylcarbamoyl substituent. AB - Various N-[3,5-bis(trifluoromethyl)benzyl]-N-methylcarbamoyl heterocycles (1, 2 and 3) modified at rings A and B in the isoquinolone (1a) and pyrido[3,4 b]pyridine (2a) nuclei were prepared and evaluated for NK1 receptor antagonistic activities. The structure-activity relationship studies on this series, along with conformational analysis, showed that (i) for ring A, 6-membered heterocycles are preferable to 5-membered heterocycles (a ca. 300-fold difference in potency), (ii) the 6-membered ring seems to function as an anchor by fixing the pendant phenyl group in a desirable orientation for receptor binding, and (iii) since compounds with aromatic rings (2) and those with aliphatic rings (3) as ring B both show good potency, this ring does not seem to be essential for receptor recognition. Among the compounds synthesized, the tetrahydropyridine derivatives 3a, 3b and 3f exhibited excellent inhibitory effects both in vitro and in vivo, with potent activity upon oral administration (ED50 = 0.20-0.27 mg/kg) (capsaicin induced plasma extravasation in guinea pig trachea). PMID- 9353895 TI - Enhanced cytotoxicity of some triterpenes toward leukemia L1210 cells cultured in low pH media: possibility of a new mode of cell killing. AB - Several triterpenes were tested for cytotoxicity by our contrived primary screening method using resting or dormant leukemia L1210 cells after 3 d preculture without medium change. Some triterpenes were found to be more cytotoxic toward the 3 d-precultured resting cells than toward the growing cells in a fresh medium. These triterpenes are distinguished by highly selective cytotoxicity toward the starved resting cells unlike common anticancer agents. The highest selectivity was shown by betulinic acid, the ratio of its IC50 values toward the growing versus resting cells amounting to 175. It is suggested that this selective cytotoxicity is attributable to low pH (< or = 6.8) of the medium. It is noteworthy that betulinic acid is not cytotoxic at all in media of ordinary pH (> or = 7.0) even after a 48-h exposure. Betulinic acid might be promising as an antitumor agent toward solid tumors because the interior pH of tumor tissues is generally lower than in normal tissues. PMID- 9353894 TI - 2-(3-Pyridyl)thiazolidine-4-carboxamides. 1. Novel orally active antagonists of platelet-activating factor (PAF). AB - In a search for novel platelet-activating factor (PAF) antagonists, we found that 1-(3-phenylpropyl)-4-[2-(3-pyridyl)thiazolidine-4-carbonyl] piperazine (3x) showed in vitro and in vivo PAF-antagonistic activities. Introduction of functional groups at the benzylic methylene carbon of 3x afforded some compounds with more potent PAF-antagonistic activity than 3x. Among them 1-(3-methyl-3 phenylbutyl)-4-[2-(3-pyridyl) thiazolidine-4-carbonyl]-piperazine fumarate (3al, YM264) was found to be one of the most potent PAF antagonists. PMID- 9353896 TI - Medicinal foodstuffs. IX. The inhibitors of glucose absorption from the leaves of Gymnema sylvestre R. BR. (Asclepiadaceae): structures of gymnemosides a and b. AB - Although the glycosidic fraction from the dried leaves of Gymnema sylvestre R. BR., gymnemic acid, was reported to be effective for diabetes, it showed little inhibitory activity on the increase of serum glucose level in oral glucose-loaded rats. From the glycosidic fraction, six triterpene glycosides, gymnemosides a, b, c, d, e, and f, were isolated together with nine known triterpene glycosides. The structures of gymnemosides a and b were determined on the basis of chemical and physicochemical evidence as 21-O-tigloyl-22-O-acetylgymnemagenin 3-O-beta-D glucopyranosiduronic acid and 16-O-acetyl-21-O-tigloylgymnemagenin 3-O-beta-D glucopyranosiduronic acid, respectively. In addition, an acetyl group linked to the 16- or 22-hydroxyl group in gymnemosides a and b was found to migrate easily to the primary 28-hydroxyl group, while the acyl migration from the 28-position was rarely observed. The inhibitory activity of each triterpene glycoside from gymnemic acid was examined to determine its impact on the increase of serum glucose level in oral glucose-loaded rats. Gymnemoside b and gymnemic acids III, V, and VII were found to exhibit a little inhibitory activity against glucose absorption, but the principal constituents, gymnemic acid I and gymnemasaponin V, lacked this activity. PMID- 9353897 TI - Estimation of surface state of poly(ethylene glycol)-coated liposomes using an aqueous two-phase partitioning technique. AB - Poly(ethylene glycol)-coated liposomes (PEG-liposomes) were prepared from distearoylphosphatidylcholine (DSPC)/cholesterol (Ch) (1:1, molar ratio) with various amounts of distearoyl-N-(monomethoxy poly(ethylene glycol)succinyl)phosphatidylethanolamine (DSPE-PEG). Surface potentials of PEG liposomes showed negative values, however, the zeta potentials were almost neutral under physiological conditions (150 mM NaCl). Taking these electrical surface properties into consideration, a non-charge-sensitive phase system consisting of 5% PEG8000 and 5% dextran T-500, 0.01 M sodium phosphate, 0.15 M sodium chloride (pH 7.0) was used to estimate the alteration of surface state of PEG-liposomes after interaction with plasma in vitro and in vivo. PEG-liposomes showed increased partitioning to the PEG phase with increasing amount of DSPE PEG. One hundred percent partitioning to the PEG phase was obtained when 2 or 1 mol% of DSPE-PEG1K or 2K was incorporated into the liposomes, respectively. This PEG/lipid ratio (mol/mol) thus afforded complete protection of the liposomal surface by the PEG moiety. When these PEG-liposomes were incubated with plasma protein (in vitro) or were recovered from liposome-injected mice (in vivo), they showed decreased partitioning to the PEG phase. However, when the in vivo-treated PEG-liposomes were purified by column chromatography and ultracentrifugation, their partitioning to the PEG phase was restored to that of PEG-liposomes incubated in phosphate-buffered saline. Thus although PEG acts as a steric barrier against the attachment of plasma protein to the liposome surface and slows down liposome clearance from the circulation in vivo, a weak interaction remains between PEG-liposome and plasma protein when the incorporated amount of DSPE-PEG is low. PMID- 9353898 TI - Comparison of nicotinamide, ethylurea and polyethylene glycol as carriers for nifedipine solid dispersion systems. AB - The most prevalent means for producing solid dispersions of nifedipine, a poorly water-soluble drug, are the solvent based processes that bring problems of environmental and health. We have investigated the preparation of solid dispersions of nifedipine (mp 173 degrees C) by the fusion method, using nicotinamide, ethylurea, polyethylene glycol (PEG) 6000 and hydroxypropylmethylcellulose (HPMC) as carriers. All these solid dispersions were obtained by cooling at room temperature after heating at 140 degrees C for 15 min. As a single carrier, nicotinamide, ethylurea and PEG were used because nifedipine dissolved in their fused liquids. Compared with the physical mixtures, the solid dispersions with ethylurea or PEG led to a higher dissolution rate of the drug, whereas the difference in drug release between the physical mixtures and the solid dispersions with nicotinamide was not clear. This peculiarity might be due to the high solubilizing effect of nicotinamide for the drug. The fused mixtures of nicotinamide-, ethylurea- or PEG-HPMC were utilized as combined carriers. HPMC dissolved in the fused liquids of nicotinamide or ethylurea, which was effective in forming the amorphous nifedipine in solid dispersions. This resulted in not only the enhanced dissolution rate but also the supersaturation behavior of nifedipine. Further, for the nicotinamide-HPMC system, the supersaturation level of nifedipine increased with an increase in the HPMC content of solid dispersions. Nicotinamide was more applicable than ethylurea and PEG for preparation of the fused dispersions of nifedipine. PMID- 9353899 TI - Fruiting-inducing activity and antifungal properties of lipid components in members of Annelida. AB - Fruiting-inducing activity and antifungal properties of lipid components in the phylum Annelida were examined. Some amphoteric cerebrosides carrying a phosphocholine group showed fruiting-inducing activity on Schizophyllum commune, and one of them possessed activity comparable to that of Sch II, the most potent substance known. Furthermore, alkyl lysophosphatidylcholines were found to have an inhibitory effect on the growth of phytopathogenic fungi, Alternaria kikuchiana and Phomopsis mali. The relationship between structure and biological activities is discussed. PMID- 9353900 TI - Structure of stratum corneum lipids characterized by FT-Raman spectroscopy and DSC. II. Mixtures of ceramides and saturated fatty acids. AB - Fourier transform (FT) Raman spectroscopy and differential scanning calorimetry (DSC) were used to study the thermotropic phase behaviour of mixtures of ceramides type IV (CER) and stearic acid (SA). For comparison the melting behaviour of SA was re-examined. The Raman spectra of all mixtures in the solid state show sharp bands associated with trans sequencies of the alkyl chain residues of both lipids. These features demonstrate that the hydrocarbon chains are highly ordered in the mixtures, too. The temperature dependence of the conformationally sensitive bands is used to estimate the degree of order in terms of the relative population of trans and gauche conformations. The DSC heating curves for the mixtures show two endothermic transitions which are typical for eutectic melting. The factor group splitting of the CH2 scissoring mode, arising from the orthorhombic subcell packing of SA, disappears in the course of the eutectic melting of samples with a SA content lower than 90 mol%. Both DSC and Raman spectroscopic studies reveal that CER and SA are immiscible in the solid state. The phase diagram of the system is a simple eutectic type one. The addition of SA to CER shifts the melting temperature of ceramides to lower values. However, though SA is a major component of stratum corneum (SC) it is not efficient enough to increase the fluidity of ceramides. PMID- 9353901 TI - Activity of phosphatidylinositol-specific phospholipase C from Bacillus cereus with thiophosphate analogs of dimyristoylphosphatidylinositol. AB - Phosphatidylinositol-specific phospholipase C (PI-PLC) was studied with sonicated dispersions of a thiophosphate analog of phosphatidylinositol, 1, 2 dimyristoyloxypropane-3-thiophospho(1D-1-myo-inositol) (D-thio-DMPI). Kinetic parameters were derived from the rate as a function of bulk lipid concentration at constant saturating surface concentration of substrate (case I), and as a function of surface concentration of substrate at a constant saturating bulk concentration of lipid (case II). The substrate, D-thio-DMPI, was diluted with L thio-DMPI or dimyristoyl phosphatidylmethanol (DMPM). In the presence of L-thio DMPI, values for Vmax = 133 mumol min-1 mg-1, Ks' (the apparent dissociation constant for the enzyme-interface complex) = 0.097 mM, and Km* (the apparent interfacial Michaelis constant) = 0.22 mol fraction were obtained. DMPM caused enzyme inhibition in case I but no inhibition in case II. L-Thio-DMPI is an ideal neutral diluent with which to study the kinetics of PI-PLC. PMID- 9353902 TI - Synchronized changing of transinterface pressure, bubble radius and surface tension: a unique feature of lung surfactant. AB - The pulsating bubble surfactometer has been commonly used to measure the minimum surface tension of lung surfactant. The complexity of the original transinterface pressure tracings and its possible physiological meanings remain undefined. In the present study, we compared surface properties between calf lung surfactant extract (CLSE) and Tween 20, a nonionic surfactant, with the pulsating bubble surfactometer. A synchronized change between transinterface pressure (P) and bubble radius (R) was observed when CLSE was tested. Mathematical analysis and computer simulation indicate that this is due to the extremely potent surface tension lowering and adjusting abilities, which allows the surface tension to decrease towards zero at the end of compression and increase towards a high surface tension during re-expansion. In contrast, a time delay between P and R was observed when Tween 20 was assessed. Surface tension adjusting ability was shown only at concentrations below or around the critical micelle concentration (cmc) of Tween 20. Surface tension became unchangeable when concentrations were further increased, suggesting amphipathic molecules were saturated on the interface. The synchronization of transinterface pressure, alveolar radius and surface tension may play an important role in maintaining the pulmonary compliance in vivo. This unique feature, observed at concentrations several orders above the cmc of phospholipids, suggests that the structure of lung surfactant at the air-liquid interface differs from that of Tween 20, a monolayer of free amphipathic molecules. PMID- 9353903 TI - Skin biopsy as a nondestructive tool for the toxicological assessment of endangered populations of pinnipeds: preliminary results on mixed function oxidase in Otaria flavescens. AB - The aim of this study was to develop and validate a method for assessing the toxicological risk of endangered populations of pinnipeds based on a nondestructive biological tool, the skin biopsy specimen. Skin biopsies can be obtained from pinnipeds by anaesthetising the animals and taking a small amount of skin in the anterior flipper area, or by shooting a biopsy dart with a crossbow. Skin biopsy material is suitable for a wide range of chemical and biomarker analysis. Organochlorines and polycyclic aromatic hydrocarbons can be analysed in subcutaneous fat and MFO activity (BPMO), Cyt.P450 isoforms, and DNA damage can be detected in epidermis. PMID- 9353904 TI - Microbial response to repeated applications of low concentrations of pentachlorophenol in an alfisol under pasture. AB - Columns of an Alfisol under permanent pasture were polluted by repeated additions of pentachlorophenol (PCP) (7 mg l-1) to levels of 102 and 510 mg Kg-1, to simulate a dynamic diffuse pollution. PCP was rapidly sorbed to the soil organic matter, and was only slightly degraded. Measurements of soil microbial biomass-C revealed a 25% decrease in total biomass-C caused by both leaching and PCP toxicity. Microbial biomass-C measurements performed on soil fractions showed that only microorganisms located in the outer compartment of the aggregates were affected. Microorganisms protected by soil micro-aggregates were not affected, suggesting that they were not in contact with PCP, which was thus unavailable for biodegradation. Three gram negative bacterial strains (Si, C3 and C2), able to use PCP as a sole carbon and energy source, were isolated after 0, 1 and 3 months of PCP enrichment respectively, and were identified as Pseudomonas (Si) and Acinetobacter (C3 and C2). In liquid degradation tests, the strains C2 and C3 degraded 60% of PCP within 26 days whereas the Pseudomonas degraded only 25%. A specific immuno-labeling of the three strains permitted to show that repeated PCP additions to soil had a positive, negative or absence of effect on the populations C2, C3 and Si respectively. PMID- 9353906 TI - A note on the use of the CEC L-33-A-93 test to predict the potential biodegradation of mineral oil based lubricants in soil. AB - The biodegradabilities of five unformulated mineral oils (brightstock, 150 SN base oil, white oil and two gas oils) were determined in the CEC L-33-A-93 test and during 20 weeks incubation in nutrient-supplemented soil microcosms. Biodegradation in both studies was measured as the loss of extractable hydrocarbon ('primary' biodegradation). There was a statistically significant (P < 0.01) rectilinear relationship between the extents of biodegradation in both test systems. The results indicate that the CEC method could be used as a relatively simple, quick and inexpensive test for assessing the potential biodegradation of mineral oil based lubricants in soil. PMID- 9353905 TI - Studies on the precursors of strong mutagen [3-chloro-4-(dichloromethyl)-5 hydroxy-2(5H)-furanone]MX by chlorination of fractions from different waters. AB - The strong mutagen, [3-chloro-4-(dichloromethyl)-5-hydroxy- 2(5H)-furanone] MX, was found to be one of the most potent mutagens in drinking water. In this study, dissolved organic matters from river water and lake water were separated into several compound classes by sorbtion on a series of resin absorbents. After chlorine treatment of the fractions, MX was determined with GC/MS in the selected ion monitoring mode. Humic substances produced more MX on a TOC-basis than other fractions and contributed more to MX formation in the chlorinated natural waters. Some phenols were detected in the oxidation products of humic substances and therefore formation of MX may occur when some phenolic precursor structures in humic substances are treated with chlorine. PMID- 9353907 TI - Bioconcentration, elimination and metabolism of 2,4-dinitrotoluene in carps(Cyprinus Carpio L.). AB - Bioconcentration curves of 2,4-dinitrotoluene(2,4-DNT) in carps (whole fish, liver, intestine and muscle) were investigated using semistatic system. For whole fish, its curve could be described as a gentle peak which began with a rise in concentration to summit or steady state, then declined and reached lower level followed by another steady state. For liver and intestine, their curves both contained two successive peaks, with the second peak followed by slight fluctuation. Bioconcentration factors of 2,4-DNT in whole fish during the first and second steady state were 9.15 and 4.15,(97.86 and 44.39, based on lipid content), respectively. By logarithmic plotting, two straight-lines with different slopes(3.6 and 0.1 d-1) were measured for elimination. According to peaky curves of 2,4-DNT in whole fish, liver and intestine, smaller BCFs than calculated BCFs based on the regression equations for inert chemicals, and large rate constants of elimination, biotransformation was inferred to have happened in tissues such as liver, intestine, and other tissues. Two metabolites were separated from liver and identified as 4-amino-2-nitrotoluene(4A2NT) and 2,4 diamino-toluene(2,4-DAT) on HPLC, their retention times were 23.1 and 8.8 min, respectively. In bioconcentration test of 2,4-DNT in liver, two metabolites and parent were determined at the same time at intervals, higher concentrations of 4A2NT and 2,4-DAT were found when level of 2,4-DNT declined. Such results demonstrated our inference that metabolism caused the declines in bioconcentration curves. A one-compartment model was set up to simulate the bioconcentration, in which biotransformation adhered to Delayed Enzyme-Catalytic Logarithmic Kinetics. Good fit of model curves with measured values could be observed. PMID- 9353908 TI - Field evidence of secondary poisoning of foxes (Vulpes vulpes) and buzzards (Buteo buteo) by bromadiolone, a 4-year survey. AB - This paper presents the result of a 4 year survey in France (1991-1994) based on the activity of a wildlife disease surveillance network (SAGIR). The purpose of this study was to evaluate the detrimental effects of anticoagulant (Ac) rodenticides in non-target wild animals. Ac poisoning accounted for a very limited number of the identified causes of death (1-3%) in most species. Predators (mainly foxes and buzzards) were potentially exposed to anticoagulant compounds (especially bromadiolone) via contaminated prey in some instances. The liver concentrations of bromadiolone residues were elevated and species-specific diagnostic values were determined. These values were quite similar to those reported in the literature when secondary anticoagulant poisoning was experimentally assessed. PMID- 9353909 TI - Potential for secondary poisoning and biomagnification in marine organisms. AB - For selected priority pollutants, like organochlorine pesticides, PAHs and PCBs, and mercury and cadmium, the transfer along marine food chains was assessed based on monitoring data. Comparison of the acquired body burden for marine fish and the toxicity thresholds for predating marine birds and mammals provides evidence for the relevance of contaminant uptake with the food and the liability for secondary poisoning. As a consequence, contaminant residues in prey organisms (critical body burden) should be used for marine hazard and risk assessments. Evaluations solely from aquatic exposure concentrations are not adequate to account for potential secondary effects in marine ecosystems. PMID- 9353910 TI - Cytochromes P450, P420 & mixed-function oxidases as biomarkers of polychlorinated biphenyl (PCB) exposure in harbour seals (Phoca vitulina). AB - Hepatic microsomal cytochrome P450, EROD and ECOD activity were investigated as biomarkers of PCB exposure in harbour seals (Phoca vitulina). Due to the difficulty of obtaining undegraded seal liver samples, standard spectrophotometric methodology was adapted to investigate P420 (degraded P450) as a PCB biomarker with partially degraded samples. Total PCB burdens in both blubber and liver had positive correlations with P450, P420 and MFO activity levels. The use of P420 biomarkers in this study supports the inclusion of samples from by-caught marine mammals for future biomonitoring studies. P450 isozymes CYP1A (P4501A) and CYP2B (P4502B) in conjunction with MFO activity were investigated as "specific" biomarkers of PCB exposure. They were found to reliably reflect levels of [MC] and [PB]-type PCB exposure in harbour seal liver. PMID- 9353911 TI - Assessment of baseline levels of PCDD/F in soils in the neighbourhood of a new hazardous waste incinerator in Catalonia, Spain. AB - In order to determine the baseline contamination by polychlorinated dibenzo-p dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) in an area from Catalonia (Spain), which will be under the influence of a new hazardous waste incinerator, PCDD/F concentrations were measured in 40 soil samples collected in the vicinity of the facility now under construction. Thirty samples represented rural soils and 10 samples urban soils. PCDD/F concentrations ranged between 0.08-8.4 ng I TEQ/kg for rural soils and 0.63-24.2 ng I-TEQ/kg for urban soils. The mean value for the 40 samples was 1.64 ng I-TEQ/kg (dry matter). The current PCDD/F levels and profiles are comparable to those found in soils from other countries. The PCDD/F concentrations found in this study show that at present the contamination by PCDD/F in soils of the examined area is rather low. PMID- 9353912 TI - Toxicity assessment of xenobiotic contaminated groundwater using lux modified Pseudomonas fluorescens. AB - A bacterial bioassay, suitable for rapid screening to assess the relative toxicity of xenobiotic contaminated groundwater has been developed. The quantitative bioassay utilizes a decline in luminescence of the lux marked soil bacterium Pseudomonas fluorescens on exposure to contaminated groundwaters from which effective concentration (EC) values can be assessed and compared. P. fluorescens was most sensitive to semivolatile organics in groundwaters but there was no correlation between EC value and chemical content. The sensitivity and reproducibility of the P. fluorescens bioassay was compared with that of Microtox and results showed that mean EC50 values for diluted ground water replicate samples were 20% and 18% respectively. This suggested that the P. fluorescens bioassay was as applicable to groundwater screening as the widely used Microtox bioassay. PMID- 9353913 TI - The bacterial flagellin gene as a biomarker for detection, population genetics and epidemiological analysis. PMID- 9353914 TI - Low-resolution sequencing of Rhodobacter sphaeroides 2.4.1T: chromosome II is a true chromosome. AB - The photosynthetic bacterium Rhodobacter sphaeroides 2.4.1T has two chromosomes, CI (approximately 3.0 Mb) and CII (approximately 0.9 Mb). In this study a low redundancy sequencing strategy was adopted to analyse 23 out of 47 cosmids from an ordered CII library. The sum of the lengths of these 23 cosmid inserts was approximately 495 kb, which comprised approximately 417 kb of unique DNA. A total of 1145 sequencing runs was carried out, with each run generating 559 +/- 268 bases of sequence to give approximately 640 kb of total sequence. After editing, approximately 2.8% bases per run were estimated to be ambiguous. After the removal of vector and Escherichia coli sequences, the remaining approximately 565 kb of R. sphaeroides sequences were assembled, generating approximately 291 kb of unique sequences. BLASTX analysis of these unique sequences suggested that approximately 131 kb (45% of the unique sequence) had matches to either known genes, or database ORFs of hypothetical or unknown function (dORFs). A total of 144 strong matches to the database was found; 101 of these matches represented genes encoding a wide variety of functions, e.g. amino acid biosynthesis, photosynthesis, nutrient transport, and various regulatory functions. Two rRNA operons (rrnB and rrnC) and five tRNAs were also identified. The remaining 160 kb of DNA sequence which did not yield database matches was then analysed using CODONPREFERENCE from the GCG package. This analysis suggested that 122 kb (42% of the total unique DNA sequence) could encode putative ORFs (pORFs), with the remaining 38 kb (13%) possibly representing non-coding intergenic DNA. From the data so far obtained, CII does not appear to be specialized for encoding any particular metabolic function, physiological state or growth condition. These data suggest that CII contains genes which are functionally as diverse as those found on any other bacterial chromosome and also contains sequences (pORFs), which may prove to be unique to this organism. PMID- 9353915 TI - Transcriptional control of several aerobically induced cytochrome structural genes in Rhodobacter sphaeroides. AB - To decipher how the synthesis of energy-transducing enzymes responds to environmental cues, the response of three Rhodobacter sphaeroides aerobic cytochrome gene promoters was analysed under different conditions. Two of these promoters are upstream of structural genes (ctaD and coxII) for individual subunits of the cytochrome aa3 respiratory complex. The third promoter is that for the cycFG operon, which encodes two c-type cytochromes of unknown function, cytochrome c554 and CycG. Primer extension analysis identified a single oxygen responsive transcription start site for each gene. Utilizing operon fusions to Escherichia coli lacZ as a measure of promoter activity, transcription from the ctaD, coxII and cycFG promoters was approximately twofold higher when cells were grown at high (30%) oxygen tensions than under low (2%) oxygen or anaerobic (photosynthetic) conditions. Analysis of promoter function using specific host mutations indicated that loss of the R. sphaeroides FNR homologue, FnrL, causes a small, but reproducible, increase in cycFG and coxII transcription when cells are grown at 2% oxygen. However, neither the delta FnrL mutation nor alterations in sequences related to a consensus target site for the E. coli FNR protein increased function of any of these three promoters to that seen under aerobic conditions in wild-type cells. From this we conclude that FnrL is not solely responsible for reduced transcription of these three aerobic cytochrome genes under low oxygen or anaerobic conditions. When activity of these three promoters was monitored after cells were shifted from anaerobic (photosynthetic) conditions to a 30% oxygen atmosphere, it took several cell doublings for LacZ levels to increase to those found in steady-state 30% oxygen cultures. From these results, it appears that activity of these promoters is also regulated by a stable molecule whose synthesis or function responds slowly to the presence of high oxygen tensions. PMID- 9353916 TI - Disruption of the Pseudomonas aeruginosa dipZ gene, encoding a putative protein disulfide reductase, leads to partial pleiotropic deficiency in c-type cytochrome biogenesis. AB - The Pseudomonas aeruginosa dipZ gene has been cloned and sequenced. Whereas disruption of Escherichia coli dipZ (dsbD), the hydrophilic C-terminal domain of which has been deduced to be periplasmic and to function as a protein-disulfide reductase, leads to the absence of c-type cytochromes, disruption of P. aeruginosa dipZ attenuated, but did not abolish, holo-c-type cytochrome biosynthesis. Comparison of the P. aeruginosa DipZ sequence with three other DipZ sequences indicated that there are not only two conserved cysteine residues in the C-terminal hydrophilic domain, but also two more in the central highly hydrophobic domain. The latter would be located toward the centre of two of the eight membrane-spanning alpha-helices predicted to compose the hydrophobic central domain of DipZ. Both these cysteine residues, plus other transmembrane helix residues, notably prolines and glycines, are also conserved in a group of membrane proteins, related to Bacillus subtilis CcdA, which lack the N- and C terminal hydrophilic domains of the DipZ proteins. It is proposed that DipZ of P. aeruginosa and other organisms transfers reducing power from the cytoplasm to the periplasm through reduction and reoxidation of an intramembrane disulfide bond, or other mechanism involving these cysteine residues, and that this function can also be performed by B. subtilis CcdA and other CcdA-like proteins. The failure of dipZ disruption to abolish c-type cytochrome synthesis in P. aeruginosa suggests that, in contrast to the situation in E. coli, the absence of DipZ can be compensated for by one or more other proteins, for example a CcdA-like protein acting in tandem with one or more thioredoxin-like proteins. PMID- 9353917 TI - A geographically widespread plasmid from Thiobacillus ferrooxidans has genes for ferredoxin-, FNR-, prismane- and NADH-oxidoreductase-like proteins which are also located on the chromosome. AB - During a search for genes encoding electron transport proteins from a Thiobacillus ferroxidans ATCC 33020 gene bank, a 19.8 kb plasmid, pTF5, which conferred increased sensitivity to the antimicrobial agent metronidazole upon an Escherichia coli mutant, was isolated and cloned in E. coli. The plasmid had an identical restriction enzyme map to a plasmid which has been found in T. ferrooxidans strains isolated from many different parts of the world. The plasmid was present at between two and four copies per genome and contained a region of approximately 5-6 kb which was also found on the chromosome. This region was sequenced and found to have four complete ORFs, which when translated had high percentage amino acid similarity to [3Fe-4S,4Fe-4S] ferredoxins, proteins of the FNR regulator family, prismane-like proteins and the NADH oxidoreductase subunit of a methane monooxygenase. In vitro protein analysis using an E. coli-derived transcription-translation system indicated that three of the four products (FdxA, PsmA and RedA) were expressed in the heterologous system. Ferredoxins, prismane like proteins and NADH oxidoreductases are redox-active proteins and it is likely that the proteins on pTF5 represent an electron transport system of as yet unknown function. Surprisingly, although genes for redox-active proteins have been isolated from other bacteria by screening gene banks for increased sensitivity to metronidazole, the region of pTF5 containing the genes for these proteins was not responsible for the increase in metronidazole sensitivity conferred by the plasmid. The region of pTF5 which did confer increased metronidazole sensitivity to an E. coli metronidazole-resistant mutant was a 319 bp region of DNA close to the origin of plasmid replication. This region contained no ORFs and was identical to that previously reported for the replicon of a 9.8 kb T. ferrooxidans plasmid, pTF191. PMID- 9353918 TI - Structural analysis of the 6 kb cryptic plasmid pFAJ2600 from Rhodococcus erythropolis NI86/21 and construction of Escherichia coli-Rhodococcus shuttle vectors. AB - The complete nucleotide sequence of the 5936 bp cryptic plasmid pFAJ2600 from Rhodococcus erythropolis NI86/21 was determined. Based on the characteristics of its putative replication genes, repA and repB, pFAJ2600 was assigned to the family of pAL5000-related small replicons identified in Mycobacterium (pAL5000), Corynebacterium (pXZ10142), Brevibacterium (pRBL1), Bifidobacterium (pMB1) and Neisseria (pJD1). The replication systems of these plasmids show striking similarities to the ones used by the ColE2 family of plasmids from Enterobacteria with respect to both trans-acting factors and ori sequences. Two possible plasmid stabilization systems are encoded on pFAJ2600: a site-specific recombinase (PmrA) related to the Escherichia coli Xer proteins for plasmid multimer resolution and an ATPase (ParA) related to the A-type of proteins in sop/par partitioning systems. The proposed replication termination region of pFAJ2600 has features in common with the Ter loci of Bacillus subtilis. Chimeras composed of a pUC18-Cmr derivative inserted in the parA-repA intergenic region of vector pFAJ2600 produced vectors that could be shuttled between Escherichia coli and several Rhodococcus species (R. erythropolis, R. fascians, R. rhodochrous, R. ruber). The pFAJ2600-based shuttle vector pFAJ2574 was stably maintained in R. erythropolis and R. fascians growing under non-selective conditions. PMID- 9353919 TI - Escherichia coli is unable to produce pyrroloquinoline quinone (PQQ). AB - Many bacteria can synthesize the cofactor pyrroloquinoline quinone (PQQ), a cofactor of several dehydrogenases, including glucose dehydrogenase (GCD). Among the enteric bacteria, Klebsiella pneumoniae has been shown to contain the genes required for PQQ biosynthesis. Escherichia coli and Salmonella typhimurium were thought to be unable to synthesize PQQ but it has been reported that strain EF260, a derivative of E. coli FB8, can synthesize PQQ after mutation and can oxidize glucose to gluconate via the GCD/PQQ pathway (F. Biville, E. Turlin & F. Gasser, 1991, J Gen Microbiol 137, 1775-1782). We have re-investigated this claim and conclude that it is most likely erroneous. (i) Strain EF260, isolated originally by Biville and coworkers, was unable to synthesize a holo-enzyme GCD unless PQQ was supplied to the growth medium. No GCD activity could be detected in membrane fractions. (ii) The amount of PQQ detected in the growth medium of EF260 was very low and not very different from that found in a medium with its parent strain or in a medium containing no cells. (iii) EF260 cells were unable to produce gluconate from glucose via the PQQ/GCD pathway. (iv) Introduction of a gcd::Cm deletion in EF260, eliminating GCD, did not affect glucose metabolism. This suggested a pathway for glucose metabolism other than the PQQ/GCD pathway. (v) Glucose uptake and metabolism in EF260 involved a low-affinity transport system of unknown identity, followed most likely by phosphorylation via glucokinase. It is concluded that E. coli cannot synthesize PQQ and that it lacks genes required for PQQ biosynthesis. PMID- 9353920 TI - Glyceraldehyde-3-phosphate dehydrogenase expression in Trichoderma harzianum is repressed during conidiation and mycoparasitism. AB - A glyceraldehyde-3-phosphate dehydrogenase (gpd) cDNA was isolated from the filamentous fungus Trichoderma harzianum in the course of a search for light regulated genes in this organism. There is apparently only one copy of gpd in the T. harzianum genome, and its sequence is most similar to that of other filamentous ascomycetes. Trichoderma grows in the soil as a saprophyte or mycoparasite. A brief pulse of blue light, or nutrient depletion, induces sporulation, which is accompanied by altered patterns of abundance of specific polypeptides. Mycoparasitic development is also accompanied by changes in gene expression. The abundance of gpd mRNA decreased strongly during sporulation, and was lowest in samples consisting o mature conidiophores and conidia. When T. harzianum was grown in the presence of cell walls of the phytopathogen Rhizoctonia solani, the gpd mRNA level was much lower than in similar cultures grown on glucose. The repression of gpd, which is usually considered a constitutively expressed gene, may be part of the switch to sporulation or to the simulated mycoparasitic state. The implications of these findings for the use of gpd promoters to confer high constitutive expression are discussed. PMID- 9353921 TI - Polyethyleneimine is an effective permeabilizer of gram-negative bacteria. AB - The effect of the polycation polyethyleneimine (PEI) on the permeability properties of the Gram-negative bacterial outer membrane was investigated using Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium as target organisms. At concentrations of less than 20 micrograms ml-1, PEI increased the bacterial uptake of 1-N-phenylnaphthylamine, which is a hydrophobic probe whose quantum yield is greatly increased in a lipid environment, indicating increased hydrophobic permeation of the outer membrane by PEI. The effect of PEI was comparable to that brought about by the well-known permeabilizer EDTA. Permeabilization by PEI was retarded but not completely inhibited by millimolar concentrations of MgCl2. PEI also increased the susceptibility of the test species to the hydrophobic antibiotics clindamycin, erythromycin, fucidin, novobiocin and rifampicin, without being directly bactericidal. PEI sensitized the bacteria to the lytic action of the detergent SDS in assays where the bacteria were pretreated with PEI. In assays where PEI and SDS were simultaneously present, no sensitization was observed, indicating that PEI and SDS were inactivating each other. In addition, a sensitizing effect to the nonionic detergent Triton X-100 was observed for P. aeruginosa. In conclusion, PEI was shown to be a potent permeabilizer of the outer membrane of Gram-negative bacteria. PMID- 9353922 TI - Peptides 14VIDLL18 and 96FEAAAL101 defined as epitopes of antibodies raised against amino acid sequences of enterotoxigenic Escherichia coli colonization factor antigen I fused to Salmonella flagellin. AB - Antibodies raised against four hybrid Salmonella flagellins carrying amino acid sequences derived from the fimbrial subunit of the colonization factor I antigen (CFA/I) of enterotoxigenic Escherichia coli (ETEC), i.e. hybrid flagellins Fla I (aa 1-15), Fla II (aa 11-25), Fla III (aa 32-45) and Fla IV (aa 88-102), were not able to inhibit the in vitro binding of CFA/I-expressing ETEC bacteria to enterocyte-like Caco-2 cells. However, one of the hybrid flagellins (Fla II) was recognized by a previously described anti-CFA/I subunit mAb (S-CFA/I 17:8) which was able to block adhesion of CFA/I-expressing bacteria to Caco-2 cells and to bind to the amino acid sequences 15IDLLQ19 of the CFA/I fimbrial subunit. Pepscan analysis of antibodies raised against the hybrid flagellins Fla II and Fla IV showed that they were specific for the sequences 14VIDLL18 and 96FEAAAL101, respectively, of the CFA/I fimbrial subunit. Thus, the discrepancy in the abilities of the anti-Fla II serum and the mAb S-CFA/I 17:8 to block binding might be ascribed to their slightly different fine specificity for epitopes. PMID- 9353923 TI - Genetic identification of chemotactic transducers for amino acids in Pseudomonas aeruginosa. AB - Two chemotactic transducer genes (termed pctB and pctC) and an open reading frame (orf1) were found in the pctA-flanking region which was previously identified as a chemotactic transducer gene in Pseudomonas aeruginosa. The pctB and pctC genes encode predicted polypeptides of 629 and 632 amino acids, respectively. Overall, PctB and PctC had 81 and 75% amino acid identities with PctA, respectively. A null mutant strain PCT2, which contained a deletion in the entire pctC, orf1, pctA and pctB genes, did not show chemotaxis towards all 20 commonly occurring L amino acids. This mutant strain also failed to respond to amino acid catabolites (cadaverine, 4-aminobutyrate and putrescine) that are strong attractants for the wild-type strain PAO1. To study the role of each gene product in L-amino acid taxis, plasmids harbouring the pctC, orf1, pctA, or pctB genes were constructed and introduced into strain PCT2 by transformation. The orf1 gene did not complement the defect in chemotaxis of strain PCT2. The pctA gene restored the ability of strain PCT2 to respond to 18 L-amino acids, suggesting that PctA plays a major role in detecting L-amino acids in P. aeruginosa. The pctB and pctC genes complemented the defect in chemotaxis to only seven (Ala, Arg, Glu, Lys, Met, Tyr, Gln) and two (His, Pro) L-amino acids, respectively. PMID- 9353924 TI - Functional and genetic characterization of mcpC, which encodes a third methyl accepting chemotaxis protein in Bacillus subtilis. AB - A 3135 bp DNA segment downstream of the spl gene on the Bacillus subtilis chromosome was cloned and its nucleotide sequence determined. An open reading frame capable of encoding a putative protein of 654 amino acids with a calculated molecular mass of 72.1 kDa was identified. The deduced amino acid sequence was similar to the McpA and McpB proteins of B. subtilis. McpA and McpB encode different methyl-accepting chemotaxis proteins (MCPs). A mutant strain containing an antibiotic resistance DNA cassette inserted into the region containing the MCP like reading frame suffered a complete loss of taxis to the amino acids cysteine, proline, threonine, glycine, serine, lysine, valine and arginine. The open reading frame was designated mcpC. The wild-type and an mcpC mutant strain were analysed for their content of methylated proteins and it was found that mcpC encodes a methylated membrane protein that has previously been designated H3. These results show that mcpC encodes a third MCP in B. subtilis. The transcription start site upstream of the mcpC gene was determined by primer extension analysis and it was found to be preceded by a potential promoter sequence that is recognized by the sigma D form of RNA polymerase. The level of beta-galactosidase expressed from a transcriptional mcpC-lacZ fusion was increased threefold when cells entered the stationary phase. No beta galactosidase could be detected in a sigD genetic background. PMID- 9353925 TI - High-osmolarity signalling in Saccharomyces cerevisiae is modulated in a carbon source-dependent fashion. AB - High-osmolarity-induced expression of the small heat-shock gene HSP12 is regulated by the HOG (high-osmolarity glycerol) pathway and PKA (protein kinase A). To analyse the regulatory input of both signal transduction pathways, high salt-induced HSP12 expression in different genetic backgrounds on glucose-, ethanol- and glycerol-based culture media was examined. Upon exposure to high osmolarity stress, the kinetics of induction of HSP12 in cells growing on the non fermentable carbon sources are strikingly different from those on glucose. Derepression of HSP12 gene expression under non-stress conditions was observed in cells growing on non-fermentable carbon sources. High-salt challenge resulted in a lower induction of the HSP12 mRNA levels in ethanol-grown cells as compared to glucose-grown cells, whereas in glycerol-grown cells hardly any high-salt induction of HSP12 mRNA levels could be detected. Analysis of signalling through the HOG pathway suggested that glycerol may influence the activity of this signalling route, possible via negative feedback. Furthermore, the cellular level of PKA activity was found to have a great impact on stress-responsive gene transcription. On the basis of the data obtained it was concluded that modulation of PKA activity plays a major role in the stress response. A glucose-dependent, PKA-regulated cellular component is postulated to affect high-osmolarity-induced HSP12 expression. PMID- 9353926 TI - Sequencing and mutagenesis of genes from the erythromycin biosynthetic gene cluster of Saccharopolyspora erythraea that are involved in L-mycarose and D desosamine production. AB - The nucleotide sequence on both sides of the eryA polyketide synthase genes of the erythromycin-producing bacterium Saccharopolyspora erythraea reveals the presence of ten genes that are involved in L-mycarose (eryB) and D-desosamine (eryC) biosynthesis or attachment. Mutant strains carrying targeted lesions in eight of these genes indicate that three (eryBIV, eryBV and eryBVI) act in L mycarose biosynthesis or attachment, while the other five (eryCII, eryCIII, eryCIV, eryCV and eryCVI) are devoted to D-desosamine biosynthesis or attachment. The remaining two genes (eryBII and eryBVII) appear to function in L-mycarose biosynthesis based on computer analysis and earlier genetic data. Three of these genes, eryBII, eryCIII and eryCII, lie between the eryAIII and eryG genes on one side of the polyketide synthase genes, while the remaining seven, eryBIV, eryBV, eryCVI, eryBVI, eryCIV, eryCV and eryBVII lie upstream of the eryAI gene on the other side of the gene cluster. The deduced products of these genes show similarities to: aldohexose 4-ketoreductases (eryBIV), aldoketo reductases (eryBII), aldohexose 5-epimerases (eryBVII), the dnmT gene of the daunomycin biosynthetic pathway of Streptomyces peucetius (eryBVI), glycosyltransferases (eryBV and eryCIII), the AscC 3,4-dehydratase from the ascarylose biosynthetic pathway of Yersinia pseudotuberculosis (eryCIV), and mammalian N methyltransferases (eryCVI). The eryCII gene resembles a cytochrome P450, but lacks the conserved cysteine residue responsible for coordination of the haem iron, while the eryCV gene displays no meaningful similarity to other known sequences. From the predicted function of these and other known eryB and eryC genes, pathways for the biosynthesis of L-mycarose and D-desosamine have been deduced. PMID- 9353927 TI - Dipeptidyl aminopeptidase processing and biosynthesis of alkaline extracellular protease from Yarrowia lipolytica. AB - Alkaline extracellular protease (AEP) from Yarrowia lipolytica is synthesized as a precursor with a 157 aa prepro-region. Signal peptide cleavage was shown to occur after Ala15 by N-terminal amino acid radiosequencing of the largest intracellular AEP precursor. AEP proteolytic activity was not required for AEP processing. After a change of the putative active site Ser to Ala, inactive AEP with the same mobility on SDS-PAGE as wild-type mature AEP was secreted. The role of dipeptidyl aminopeptidase (DPAPase) activity in AEP processing was also investigated. Mutations early in the -X-Ala- and -X-Pro- dipeptide stretch (Pro17 to Met which should prevent DPAPase processing and Ala19 to Val which should allow removal of only the first dipeptide) did not prevent synthesis of active mature AEP nor did use of the DPAPase inhibitor ProboroPro. Deletion of the entire dipeptide stretch (Ala16 to Pro33) resulted in intracellular accumulation of an AEP precursor, which surprisingly was not glycosylated, and little or no secretion of AEP-related polypeptides. Expression of AEP in wild-type and dpp1 dap2 Saccharomyces cerevisiae strains (lacking both the Golgi and vacuolar DPAPases) resulted in secretion of only mature AEP and no AEP precursors. Transit times and levels of AEP secretion were similar for both strains. These results indicate that the KEX2-like cleavage after Lys156-Arg157, which yields mature active AEP can occur in the absence of DPAPase processing and that DPAPase processing is not necessary for secretion of mature active AEP. PMID- 9353928 TI - Acetate kinase from Clostridium acetobutylicum: a highly specific enzyme that is actively transcribed during acidogenesis and solventogenesis. AB - Acetate kinase (ATP:phosphotransferase, EC 2.7.2.1) has been purified 294-fold from acid-producing cells of Clostridium acetobutylicum DSM 1731 to a specific activity of 1087 U mg-1 (ADP-forming direction). The dimeric enzyme consisted of subunits with a molecular mass of 43 kDa. The molecular mass of the native acetate kinase was in the range 87-94 kDa as judged by gel filtration and native gel electrophoresis. The enzyme showed high specificity for the substrates acetate and ATP, and maximal activity was obtained with Mn2+ as divalent cation. The presence of mercury compounds such as HgCl2 and p-hydroxymercuribenzoate resulted in an essential loss of activity. The apparent K(m) values of acetate, Mg-ATP, acetyl phosphate, and Mg-ADP were 73, 0.37, 0.58 and 0.71 mM. An activity staining procedure for detection of acetate kinase in crude cell extracts after separation on native polyacrylamide gels was developed. A DNA fragment encoding 246 bp of the acetate kinase gene of C. acetobutylicum DSM 792 was cloned by a PCR-based approach. Northern blot analysis revealed transcription of the gene under acid- and solvent-producing conditions with no significant differences at the level of transcription. PMID- 9353929 TI - Molecular analysis of a Clostridium butyricum NCIMB 7423 gene encoding 4-alpha glucanotransferase and characterization of the recombinant enzyme produced in Escherichia coli. AB - An Escherichia coli clone was detected in a Clostridium butyricum NCIMB 7423 plasmid library capable of degrading soluble amylose. Deletion subcloning of its recombinant plasmid indicated that the gene(s) responsible for amylose degradation was localized on a 1.8 kb NspHI-Scal fragment. This region was sequenced in its entirety and shown to encompass a large ORF capable of encoding a protein with a calculated molecular mass of 57,184 Da. Although the deduced amino acid sequence showed only weak similarity with known amylases, significant sequences identity was apparent with the 4-alpha-glucano-transferase enzymes of Streptococcus pneumoniae (46.9%), potato (42.9%) and E. coli (16.2%). The clostridial gene (designated maIQ) was followed by a second ORF which, through its homology to the equivalent enzymes of E. coli and S. pneumoniae, was deduced to encode maltodextrin phosphorylase (MaIP). The translation stop codon of MaIQ overlapped the translation start codon of the putative maIP gene, suggesting that the two genes may be both transcriptionally and translationally coupled. 4-alpha Glucanotransferase catalyses a disproportionation reaction in which single or multiple glucose units from oligosaccharides are transferred to the 4-hydroxyl group of acceptor sugars. Characterization of the recombinant C. butyricum enzyme demonstrated that glucose, maltose and maltotriose could act as acceptor, whereas of the three only maltotriose could act as donor. The enzyme therefore shares properties with the E. coli MaIQ protein, but differs significantly from the glucanotransferase of Thermotoga maritima, which is unable to use maltotriose as donor or glucose as acceptor. Physiologically, the concerted action of 4-alpha glucanotransferase and maltodextrin phosphorylase provides C. butyricum with a mechanism of utilizing amylose/maltodextrins with little drain on cellular ATP reserves. PMID- 9353930 TI - Characterization of the rate-limiting step of the secretion of Bacillus subtilis alpha-amylase overproduced during the exponential phase of growth. AB - The Bacillus subtilis alpha-amylase gene, amyE, was expressed under the regulated control of sacR, the levansucrase leader region. The gene fusion including the complete amyE coding sequence with the signal peptide sequence was integrated into the chromosome of a degU32(Hy) strain deleted of the sacB DNA fragment. In this genetic contex, alpha-amylase is produced in the culture supernatant at a high level (2% of total protein) during the exponential phase of growth upon induction by sucrose. Pulse-chase experiments showed that the rate-limiting step (t1/2 = 120 s) of the secretion process is the release of a cell-associated precursor form whose signal peptide has been cleaved. The efficiency of this ultimate step of secretion decreased dramatically in the presence of a metal chelator (EDTA) or when the cells were converted to protoplasts. The hypothesis that this step is tightly coupled with the folding process of alpha-amylase occurring within the cell wall environment was substantiated by in vitro folding studies. The unfolding-folding transition, monitored by the resistance to proteolysis, was achieved within the same time range (t1/2 = 60 s) and required the presence of calcium. This metal requirement could possibly be satisfied in vivo by the integrity of the cell wall. The t1/2 of the alpha-amylase release step is double that of levansucrase, although their folding rates are similar. This perhaps indicates that the passage through the cell wall may depend on parietal properties (e.g. metal ion binding and porosity) and on certain intrinsic properties of the protein (molecular mass and folding properties). PMID- 9353931 TI - Sequencing of regions downstream of addA (98 degrees) and citG (289 degrees) in Bacillus subtilis. AB - The nucleotide sequence of 17.3 kbp downstream of addA (98 degrees) on the Bacillus subtilis chromosome was determined. Twenty putative ORFs were identified. Three of them coincided with known B. subtilis genes, addA, sbcD and wprA. The product of four other ORFs showed similarity to SbcC of Clostridium perfringens, CotH of B. subtilis, 2-hydroxyhepta-2,4-diene-1,7-diodate isomerase of Methanococcus jannaschi and a putative ORF of Pseudomonas syringae. In addition, a sequence of 7.6 kbp downstream of citG (189 degrees) was analysed. Among 10 putative ORFs identified, two coincided with known genes, citG and mrgA, whilst three showed homology with X86780, a sensory protein kinase of Streptomyces hygroscopicus, an alkaline phosphatase regulatory protein and a hypothetical protease, YyxA, of B. subtilis. PMID- 9353932 TI - A Bacillus subtilis chromosome segment at the 100 degrees to 102 degrees position encoding 11 membrane proteins. AB - The 25.9 kbp region upstream of nprB at 100 degrees-102 degrees on the Bacillus subtilis chromosome was sequenced. This revealed a known gene, degA, which was previously mislocated on the genetic map. A total of 29 putative ORFs were identified including a cluster of three ORFs whose products show clear homology with sulphate adenylyl pathway enzymes and, in addition, 11 ORFs whose products have one or more membrane domains, as indicated by their hydropathy profiles. PMID- 9353933 TI - The Bacillus subtilis genome from gerBC (311 degrees) to licR (334 degrees). AB - As part of the international project to sequence the Bacillus subtilis genome, the DNA region located between gerBC (311 degrees) and licR (334 degrees) was assigned to the institut Pasteur. In this paper, the cloning and sequencing of 176 kb of DNA and the analysis of the sequence of the entire 271 kb region (6.5% of the B. subtilis chromosome) is described; 273 putative coding sequences were identified. Although the complete genome sequences of seven other organisms (five bacteria, one archaeon and the yeast Saccharomyces cerevisiae) are available in public database, 65 genes from this region of the B. subtilis chromosome encode proteins without significant similarities to other known protein sequences. Among the 208 other genes, 115 have paralogues in the currently known B. subtilis DNA sequences and the products of 178 genes were found to display similarities to protein sequences from public databases for which a function is known. Classification of these genes shows a high proportion of them to be involved in the adaptation to various growth conditions (non-essential cell wall constituents, catabolic and bioenergetic pathways); a small number of the genes are essential or encode anabolic enzymes. PMID- 9353934 TI - Comparison of biological effect of the two different enterotoxin complexes isolated from three different strains of Bacillus cereus. AB - The cytotoxicity of the two different enterotoxin complexes of Bacillus cereus was compared after isolation from three different strains. Protein components of non-haemolytic enterotoxin (NHE) of 39 kDa, 45 kDa and 105 kDa were isolated from all of the three strains, whilst proteins B, L1 and L2 of haemolysin BL (HBL) were isolated from supernatants of two strains (F837-76 and 1230-88). These proteins were not detected in strain 0075-95. Inhibition of protein synthesis in Vero cells was used as a measure of cytotoxicity. The HBL complex from strain F837-76 was highly toxic. This strain also produced the NHE complex. However, when purified, at least two of the components of NHE had to be present in higher amounts than those of the components of HBL to cause the same degree of toxicity. Both complexes purified from strain 1230-88 were cytotoxic. The amount required to cause the same degree of cytotoxicity was approximately equal for the components of the two complexes, except that higher amounts of the 105 kDa protein of NHE had to be present than for the other components. None of the purified complexes from strain 1230-88 was toxic in amounts comparable to those of the HBL complex of strain F837-76 and NHE of strain 0075-95. These results indicate that when measuring cytotoxic enterotoxins from B. cereus at least two different complexes and six different proteins have to be taken into consideration. PMID- 9353935 TI - Recombinant SNAP-25 is an effective substrate for Clostridium botulinum type A toxin endopeptidase activity in vitro. AB - Bacterial neurotoxins are now being used routinely for the treatment of neuromuscular conditions. Alternative assays to replace or to complement in vivo bioassay methods for assessment of the safety and potency of these botulinum neurotoxin-based therapeutic products are urgently needed. Advances made in understanding the mode of action of clostridial neurotoxins have provided the basis for the development of alternative mechanism-based assay methods. Thus, the identification of SNAP-25 (synaptosomal-associated protein of molecular mass 25 kDa) as the intracellular protein target which is selectively cleaved during poisoning by botulinum neurotoxin type A (BoNT/A) has enabled the development of a functional in vitro assay for this toxin. Using recombinant DNA methods, a segment of SNAP-25 (aa residues 134-206) spanning the toxin cleavage site was prepared as a fusion protein to the maltose-binding protein in Escherichia coli. The fusion protein was purified by affinity chromatography and the fragment isolated after cleavage with Factor Xa. Targeted antibodies specific for the N and C termini of SNAP-25, as well as the toxin cleavage site, were prepared and used in an immunoassay to demonstrate BoNT/A endopeptidase activity towards recombinant SNAP-25 substrates. The reaction required low concentrations of reducing agents which were inhibitory at higher concentrations as were metal chelators and some inhibitors of metallopeptidases. The endopeptidase assay has proved to be more sensitive than the mouse bioassay for detection of toxin in therapeutic preparations. A good correlation with results obtained in the in vivo bioassay (r = 0.95, n = 23) was demonstrated. The endopeptidase assay described here may provide a suitable replacement assay for the estimation of the potency of type A toxin in therapeutic preparations. PMID- 9353936 TI - Production of putative virulence factors by Renibacterium salmoninarum grown in cell culture. AB - A cell culture system, employing the fish cell line Epithelioma papillosum cyprini (EPC), was developed to study the synthesis of intracellular antigen and the expression of putative virulence factors by Renibacterium salmoninarum. EPC cultures infected with R. salmoninarum could be maintained for 7 weeks, during which the pathogen multiplied intracellularly. Immunohistochemical examination of infected cultures revealed the production of the p57 antigen, haemolysin and cytolysin. The intracellular nature of the infection was confirmed by transmission electron microscopic examination of EPC monolayers. A comparison of the relative virulence of bacterial cells cultured in EPC cells and on agar plates revealed that the former were markedly more virulent in challenge experiments with juvenile rainbow trout (Oncorhynchus mykiss Walbaum). The EPC cell culture model provided a system for the study of R. salmoninarum under more natural conditions than those achieved with plate culture techniques. PMID- 9353937 TI - Evidence for Serpulina hyodysenteriae being recombinant, with an epidemic population structure. AB - The population structure of Serpulina hyodysenteriae was investigated using multilocus enzyme electrophoresis. A total of 231 isolates were divided into 50 electrophoretic types (ETs), with a mean genetic diversity of 0.29 for the number of ETs and 0.23 for the number of isolates. Subsets of isolates from two Australian states (71 isolates from Victoria and 68 isolates from Queensland) exhibited as much genetic variation as the entire collection. The calculated index of association (IA) for the number of ETs (0.29 +/- 0.17) was not significantly different from zero, and hence provided evidence for the occurrence of significant genetic recombination accounting for the observed variation between strains. In contrast, the IA for the number of isolates (3.93 +/- 0.03) was significantly different from zero, with seven of the 50 ETs (ETs 4, 6, 13, 14, 20, 33 and 35) containing 51% of all the isolates. Even when multiple isolates from the same farm were removed from the analysis, the IA value for the number of isolates remained significantly greater than zero (IA 9.87 +/- 0.04), indicating that it was not biased by their inclusion. The results suggest that S. hyodysenteriae has an epidemic population structure. PMID- 9353938 TI - The pncA gene from naturally pyrazinamide-resistant Mycobacterium avium encodes pyrazinamidase and confers pyrazinamide susceptibility to resistant M. tuberculosis complex organisms. AB - The antituberculosis drug pyrazinamide (PZA) needs to be converted into pyrazinoic acid (POA) by the bacterial pyrazinamidase (PZase) in order to show bactericidal activity against Mycobacterium tuberculosis. M. avium is naturally resistant to PZA. To investigate whether this natural resistance to PZA is due to inability of the M. avium PZase to convert PZA to bactericidal POA, the M. avium PZase gene (pncA) was cloned by using the M. tuberculosis pncA gene as a probe. Sequence analysis showed that the M. avium pncA gene is 561 bp long, encoding a protein with a predicted size of about 19.8 kDa; but Western blotting showed that the M. avium PZase migrated as a 24 kDa band when expressed in M. bovis BCG and Escherichia coli. Sequence comparison revealed that M. avium PZase has 67.7% and 32.8% amino acid identity with the corresponding enzymes from M. tuberculosis and E. coli, respectively. Southern blot analysis with the M. avium pncA gene as a probe showed that M. terrae, M. gastri, M. marinum, M. fortuitum, M. xenopi, M. gordonae, M. szulgai, M. celatum and M. kansasii have close pncA homologues, whereas M. chelonae and M. smegmatis did not give significant hybridization signals. Transformation with the M. avium pncA gene conferred PZA susceptibility to PZA-resistant M. tuberculosis complex organisms, indicating that the nonsusceptibility of M. avium to PZA is not due to an ineffective PZase enzyme, but appears to be related to other factors such as transport of POA. PMID- 9353939 TI - Targets for pSAM2 integrase-mediated site-specific integration in the Mycobacterium smegmatis chromosome. AB - An improved integrative cassette from plasmid pSAM2 has been constructed containing plasmid int and attP genes but excluding the xis gene, which should results in increased stability by suppression of the excision reaction. This cassette was included in both suicide and thermosensitive plasmids and used for integration in Mycobacterium smegmatis. Suicide plasmids containing this cassette integrated at a single site (attB1) in the M. smegmatis chromosome. The sequence of the attB1 site has been determined and was identified as a putative tRNA(Pro) gene. Thermosensitive plasmids containing the cassette integrated both at the same attB1 site and at other different sites, often giving rise to simultaneous integration at two sites. A second integration site (attB2) has been sequenced, which was located in the region encoding 16S rRNA of one of the two rrn operons of M. smegmatis. PMID- 9353940 TI - Sequence and RFLP analysis of the elongation factor Tu gene used in differentiation and classification of phytoplasmas. AB - Primers designed from sequences of the gene encoding the elongation factor Tu (tuf gene) of several culturable mollicutes amplified most of the tuf gene from phytoplasmas of the aster yellows, stolbur and X-disease groups. About 85% of the tuf gene from two aster yellows strains and a tomato stolbur phytoplasma was sequenced. The nucleotide sequence similarity between these related phytoplasmas was between 87.8 and 97.0%, whereas the homology with other mollicutes was 66.3 72.7%. The similarity of the deduced amino acid sequence was significantly higher, ranging from 96.0 to 99.4% among the phytoplasmas and 78.5% to 83.3% between phytoplasmas and the culturable mollicutes examined. From the nucleotide sequences of the phytoplasma strains, two pairs of primers were designed; one amplified the phytoplasmas of most phylogenetic groups that were established, the other was specific for the aster yellows and stolbur groups. The phytoplasmas of the various groups that were amplified could be distinguished by RFLP analysis using Sau3AI, Alul and HpaII. The aster yellows group could be divided into five Sau3AI RFLP groups. These results showed that the tuf gene has the potential to be used to differentiate and classify phytoplasmas. Southern blot analysis revealed that the tuf gene is present as a single copy. PMID- 9353941 TI - The immunoreactive 116 kDa surface protein of Mycoplasma pneumoniae is encoded in an operon. AB - Sera from 10 patients infected with Mycoplasma pneumoniae were used in Western blot analysis of Triton-X-114-soluble protein preparations of M. pneumoniae. All 10 sera were reactive with a protein antigen of 116 kDa. Sera from another 17 patients were used in Western blot analysis of whole-cell M. pneumoniae proteins; 15 of these sera were reactive with the 116 kDa protein. Trypsin digestion of whole M. pneumoniae cells demonstrated the surface location of this protein. Sequencing of DNA which contained the gene for this protein identified an ORF of 3093 bp encoding a protein with a predicted molecular mass of 116013 Da. The ORF for the 116 kDa protein had 99.8% nucleotide identity with the M. pneumoniae gene G07_orf1030 and 61% nucleotide identity with the Mycoplasma genitalium ORF MG075 of unassigned function. An ORF which was identified 5' to the 116 kDa protein ORF coded for a 16 kDa protein and had 99.8% nucleotide identity with the M. pneumoniae gene G07_orf135 and 58.4% nucleotide identity with the ORF MG074 of M. genitalium. Analysis of mRNA detected a 3.7 kb transcript with a single initiation site 5' to the ORF encoding the 16 kDa protein. The coding sequences for both the 16 kDa protein and the 116 kDa protein were present in this transcript, indicating that they were part of an operon and suggesting a possible functional relationship. PMID- 9353942 TI - The phtE locus in the phaseolotoxin gene cluster has ORFs with homologies to genes encoding amino acid transferases, the AraC family of transcriptional factors, and fatty acid desaturases. AB - A cluster of genes involved in the production of phaseolotoxin, a phytotoxin produced by Pseudomonas syringae pv. phaseolicola, contains eight (phtA through phtH) complementation groups (Y. X. Zhang, K. B. Rowley, and S. S. Patil, J. Bacteriol., 175:6451-6458, 1993). In this study, sequencing of the region encompassing the phtE locus revealed six putative open reading frames (ORFs), each preceded by a putative ribosomal binding site, and all oriented in the same direction. Reverse transcription-polymerase chain reaction suggested that the phtE locus is transcribed as one large (6.4 kb) transcript, indicating that the ORFs constitute an operon. Primer extension analysis showed that the transcript begins at a T, located 31 bp upstream of the ATG codon of ORF1. Comparison of the sequences of the putative ORFs with the sequences of known genes revealed that ORF3, encoding a protein containing 395 amino acids, has 55% similarity to the acetylornithine aminotransferase gene from Escherichia coli, and the ornithine aminotransferase genes from other organisms. A lysine residue that is a binding site for pyridoxal phosphate and an arginine residue that is a binding site for the alpha-carboxylate group of the substrate are conserved in ORF3. These data suggest that ORF3 encodes a protein involved in the biosynthesis of ornithine, a constituent of phaseolotoxin. ORF5, encoding a peptide of 378 amino acid residues, possesses a helix-turn-helix motif at the C-terminal end that is characteristic of the AraC family of transcriptional factors, and there is a possible leucine zipper at the N-terminal end of this peptide. ORF6, encoding a protein of 327 amino acids, has about 40% similarity with the fatty acid desaturase gene, desA, of Synechocystis Pcc6803 and considerable similarity with fatty acid desaturase genes from other organisms. ORF6 and desA show very similar hydropathy profiles and both contain a copper binding signature. Computer searches did not discover significant homologies in the data base for the other ORFs, but hydropathy analysis showed that all of them contain one to several hydrophobic domains, suggesting that the gene products of these ORFs may be membrane associated. PMID- 9353943 TI - An immunoreactive protein to wheat-germ agglutinin antibody is induced in oat roots following invasion of the cereal cyst nematode Heterodera avenae, and by jasmonate. AB - A protein that cross-reacts to a wheat-germ agglutinin antibody was induced in oat roots following the invasion of second-stage juveniles (J2) of the cereal cyst nematode Heterodera avenae. This protein, designated ASP45, was acid soluble, and its molecular mass was about 45 kDa on a sodium dodecyl sulfate polyacrylamide gel. ASP45 was induced in both compatible and incompatible interactions between the nematode and the plant, and also in roots by exposure to jasmonic acid (JA) or methyl jasmonate. However, ASP45 was not induced by elicitors of pathogenesis-related proteins, abscisic acid, or wounding. Lipoxygenase activity, which is involved in JA synthesis, was higher in nematode infected and JA-treated roots than in their noninfected, untreated counterparts. Inhibition of lipoxygenase activity in roots abolished ASP45 induction in the nematode-infected roots. Amino acid sequences similar to that of ASP45 were found in chitinases of poplar tree and Arabidopsis, even though ASP45 showed no chitinase activity. Although the biological role of ASP45 in infected roots is not clear, JA is suggested to be involved in signal transduction after pathogen invasion of the plant. PMID- 9353944 TI - A transgenic mutant of Lactuca sativa (lettuce) with a T-DNA tightly linked to loss of downy mildew resistance. AB - One hundred and ninety-two independent primary transformants of lettuce cv. Diana were obtained by co-cultivation with Agrobacterium tumefaciens carrying constructs containing maize Ac transposase and Ds. R2 families were screened for mutations at four genes (Dm) for resistance to downy mildew. One family, designated dm3t524, had lost resistance to an isolate of Bremia lactucae expressing the avirulence gene Avr3. Loss of resistance segregated as a single recessive allele of Dm3. The mutation was not due to a large deletion as all molecular markers flanking Dm3 were present. Loss of Dm3 activity co-segregated with a T-DNA from which Ds had excised. Genomic DNA flanking the right border of this T-DNA was isolated by inverse polymerase chain reaction. This genomic sequence was present in four to five copies in wild-type cv. Diana. One copy was missing in all eight deletion mutants of Dm3 and altered in dm3t524, indicating tight physical linkage to Dm3. Three open reading frames (ORFs) occurred in a 6.6 kb region flanking the insertion site; however, expression of these ORFs was not detected. No similarities were detected between these ORFs and resistance genes cloned from other species. Transgenic complementation with 11-to 27-kb genomic fragments of Diana spanning the insertion site failed to restore Dm3 function to two ethyl methanesulfonate (EMS)-induced mutants of Dm3 or to cv. Cobham Green, which naturally lacks Dm3 activity. Therefore, either the T-DNA inserted extremely close to, but not within, Dm3 and the mutation may have been caused by secondary movement of Ds, or Dm3 activity is encoded by a gene extending beyond the fragments used for complementation. PMID- 9353946 TI - Targeted disruption of a fungal G-protein beta subunit gene results in increased vegetative growth but reduced virulence. AB - Targeted disruption of two G-protein alpha subunit genes in the chestnut blight fungus Cryphonectria parasitica revealed roles for the Gi alpha subunit CPG-1 in fungal reproduction, virulence, and vegetative growth. A second G alpha subunit, CPG-2, was found to be dispensable for these functions. We now report the cloning and targeted disruption of a C. parasitica G-protein beta subunit gene. The deduced amino acid sequence encoded by this gene, designated cpgb-1, was found to share 66.2, 65.9, and 66.7% amino acid identity with G beta homologues from human, Drosophila, and Dictyostelium origins, respectively, but only 39.7% identity with the Saccharomyces cerevisiae G beta homologue STE4 product. Low stringency Southern hybridization failed to detect any related G beta subunit genes in C. parasitica. Targeted disruption of cpgb-1 resulted in several of the changes previously reported to accompany disruption of the C. parasitica Gi alpha subunit gene cpg-1. These included very significant reductions in pigmentation, asexual sporulation, and virulence. In contrast to results obtained for Gi alpha gene disruption, the reduction in virulence resulting from the disruption of a G beta gene was accompanied by increased, rather than decreased, vegetative growth on synthetic medium. The relevance of these results to mechanisms of fungal virulence is considered. PMID- 9353947 TI - Encapsidation of potyviral RNA in various forms of transgene coat protein is not correlated with resistance in transgenic plants. AB - Transgenic plants expressing either bean yellow mosaic potyvirus or chimeric potyvirus coat protein (CP) were inoculated with various potyviruses. Antigen coated plate, indirect enzyme-linked immunosorbent assay and immunoelectron microscopy of virus purified from transgenic plants showed that progeny virions contained from < 1% to as much as 25% transgenic CP. Different levels of transcapsidation may reflect the extent of compatibility between transgene CP and the viral CP. PMID- 9353948 TI - A novel class of elicitin-like genes from Phytophthora infestans. AB - Elicitins are a family of structurally related proteins that induce hypersensitive response in specific plant species. Two Phytophthora infestans cDNAs, inf2A and inf2B, potentially encoding novel elicitin-like proteins, were isolated from a cDNA library made from infected potato tissue. Multiple sequence alignments and phylogenetic analyses of 19 elicitins and elicitin-like proteins from nine Phytophthora spp. and from Pythium vexans suggest that there are at least five distinct classes within the elicitin family. PMID- 9353949 TI - Information processing through the first year of life: a longitudinal study using the visual expectation paradigm. AB - This Monograph uses a developmental function approach to describe age-related change and individual differences in infant information processing during the first year of life. The Visual Expectation Paradigm (VExP) is used to measure speed of information processing, response variability, and expectancy formation. Eye-movement reaction times and anticipatory saccades were gathered from 13 infants assessed monthly from 2 to 9 months and then again at 12 months. Analysis of response patterns demonstrated the applicability of the paradigm throughout the age range studied. Converging operations strongly indicate that the traditional estimate of the minimum time required for infants to initiate a saccade to a peripheral stimulus may be as much as 100 milliseconds (ms) too long. Moreover, the newly estimated minimum of 133 ms does not appear to change during the 2-12-month period. Reanalysis of the present data and past research reveals that the new, shorter minimum reaction time is unlikely to affect findings based on mean reaction time. However, using the traditional minimum reaction time will inflate estimates of percentage anticipation, especially in infants older than 5 months. Group and individual growth curves are described through quantitative models of four variables: reaction time, standard deviation of reaction time, percentage anticipation, and anticipation latency. Developmental change in reaction time was best described by an asymptotic exponential function, and evidence for a local asymptote during infancy is presented. Variability in reaction time was found to decline with age, independent of mean reaction time, and was best described by a polynomial function with linear and quadratic terms. Anticipation showed little lawful change during any portion of the age span, but latency to anticipate declined linearly throughout the first year. Stability of individual differences was strong between consecutive assessments of mean reaction time. For nonconsecutive assessments, stability was found only for the 6-12-month period. Month-to-month stability was inconsistent for reaction-time variability and weak for both anticipation measures. Analyses of individual differences in growth curves were carried out using random regressions for the polynomial models. The only significant individual difference (in growth curves) was found for reaction-time variability. Parameter estimates from the exponential models for reaction time suggested two or three developmental patterns with different exponential trajectories. This finding indicates that the strong form of the exponential growth hypothesis, which states that processing speed develops at the same rate for all individuals, does not hold for the first year of life. In the concluding chapter, Grice's Variable Criterion Model (Grice, 1968) is used to integrate three key findings: regular age changes in mean reaction time and variability but no age change in the minimum reaction time. It is argued that the rate of growth of sensory-detection information is developmentally constant during much of the first year but that age changes occur in the level and spread of the distribution of response threshold values. The unique strengths of the paradigm are discussed, and future directions are suggested for further developing the paradigm itself and for using it as a tool to study broad issues in infant cognition. PMID- 9353950 TI - Models of oculomotor variability in infancy. PMID- 9353951 TI - Infant visual expectations: advances and issues. PMID- 9353952 TI - Effects of low dose X-ray irradiation on purine metabolism in mouse splenocytes. AB - This study examines the influence of low dose X-ray irradiation on purine nucleotide metabolites such as adenosine, inosine, hypoxanthine, xanthine and uric acid, and hence generation of ATP-mediated energy in mouse splenocytes. It was found that, unlike high dose irradiation which promotes membrane damage, low dose irradiation enhances the ability to regulate the energy metabolisms as reflected by the increase in Na+, K(+)-ATPase activity and the adequate activation of the above salvage pathway. Namely, the levels of adenosine, inosine and uric acid significantly increased, while the levels of xanthine and hypoxanthine decreased significantly. Moreover, the cysteine level and superoxide dismutase activity significantly increased at a dosage of 20 cGy. PMID- 9353953 TI - Effect of glucose-cysteine adduct on cysteine desulfuration in guinea pig tissues. AB - Effect of intraperitoneal administration (12 mmol/kg of body weight) of glucose cysteine adduct 2-(D-gluco-pentahydroxypentyl)-thiazolidine-4-carboxylate, (glc cys) on the rhodanese, gamma-cystathionase and 3-mercaptopyruvate sulfurtransferase (MPST) activity levels in guinea pig tissues was studied. The rhodanese activity value in liver increased by 41%, 3-mercaptopyruvate sulfurtransferase by 24%, and gamma-cystathionase by 12% after three successive days of the administration. In the kidney, on the contrary, glc-cys administration resulted in about 18% decrease in the gamma-cystathionase activity value, whereas no changes in MPST and rhodanese activity values were observed. In the case of the brain, rhodanese and gamma-cystathionase did not change their activity but the activity of MPST decreased by 21%. MPST level did not change substantially in whole blood after glc-cys treatment. The results seem to indicate that in guinea pig liver but not in kidney and brain, glc-cys has a potential to activate the desulfuration pathway of L-cysteine metabolism. PMID- 9353954 TI - The role of peroxidase in neuromelanin synthesis: a review. AB - There is currently no general agreement on the enzymatic basis of neuromelanin synthesis. It is generally agreed that neuromelanin synthesis must differ from melanin synthesis in skin, since "tyrosinase" (aerobic dopa oxidase) is not present in brain. Various proposals have suggested that monoamine oxidase, prostaglandin H synthetase, or peroxidase may catalyze neuromelanin formation from dopa, dopamine and norepinephrine. It has also been proposed that neuromelanin synthesis may be based on the pseudoperoxidase activity of metal ions. Our group was the first to demonstrate enzymatic peroxidase activity in brain, which was subsequently confirmed by three other laboratories. Our group was also the first to demonstrate the melanogenic activity of brain peroxidase (subsequently confirmed by another laboratory) and to propose that peroxidase is the key enzyme in neuromelanin synthesis because of this potential and because of its presence in lysosomes, the subcellular site of neuromalin. PMID- 9353955 TI - Acetate or octanoate increases glycogenolysis in smooth muscle as determined by 13C-NMR. AB - Vascular smooth muscle is considered a model system for the study of the compartmentation of carbohydrate metabolism. Since vascular smooth muscle is capable of synthesizing substantial glycogen stores and utilizing a variety of metabolic substrates, we sought to determine the effects of utilization of either acetate or octanoate on 2-13C-glucose metabolism and on 1-13C-glycogen metabolism in contracting hog carotid arteries having high levels of 13C-labeled glycogen. In carotid arteries that have been allowed to synthesize substantial amounts of glycogen (to a total content of 7.84 +/- 0.31 mumol/gm wet wt glucosyl units), provision of either 5 mM 2-13C-glucose and 2 mM sodium acetate or 5 mM 2-13C glucose and 0.5 mM octanoic acid during a 3 hr. contraction resulted in a 74% and 71% increase in glycogen utilization compared to that in the presence of glucose as the sole exogenous substrate. The fraction of the 3-13C-lactate from 1-13C glycogen was substantially reduced in the presence of either acetate or octanoate. We speculate that AMP production by thiokinases with localized access of AMP to phosphorylase may be the common mechanism of glycogenolytic modulation by acetate and octanoate in contracting vascular smooth muscle with high glycogen content. We conclude that utilization of exogenous fatty acids can regulate glycogenolysis independent of glycolysis in contracting vascular smooth muscle with high glycogen content. PMID- 9353957 TI - NMR relaxation time studies of large bowel neoplasms. AB - The purpose of the study was to determine if measurements of the NMR relaxation times could be useful in distinguishing healthy from neoplasmatic tissue in in vitro study of the slices from the large bowel obtained during surgery before histological examination. Tissue samples taken from the center of the tumor and from the distal part of the excised large bowel segment were stored at 4 degrees C in closed tubes not longer than 24 hours. The measurements were performed at 37 degrees C using a pulse spectrometer operating at 27 MHz. After NMR investigation all the tissue samples were preserved and carefully examined by conventional histopathological techniques. Both for the T1 and the T2 the mean values of the relaxation time for the respective neoplasmatically changed group and normal, non changed group, were statistically different. However, the differences in measured relaxation times are too small to use the NMR method alone for diagnosis. The NMR method can be useful only for an initial selection of tissue samples with possible cancer changes but histopathological examination is required to verify that cancer is present in the tissue. PMID- 9353956 TI - Local changes in membrane potential intensify neutrophil oxidative burst. AB - The aim of this paper is to study the effects of the pulsed electrical field/current alone or combined with ionomycin, fMLP and PMA, the chemical stimuli that operate through distinctly different activation pathways, on the time course of the oxidative burst response in human neutrophils. Neither the control groups nor the neutrophils treated with electrical field alone showed any increase in oxidative burst activity measured by the luminol-enhanced chemiluminescence technique. It was found that electrical treatment potentiates chemically induced activation with either of the chemical stimulators used. The integrated oxidative burst response--which represents a cumulative amount of oxygen metabolites produced during whole response--was 87% higher in neutrophils treated with a combination of ionomycin and an electric field than in solely ionomycin treated cells, while the peak level of the response was 114% higher. In neutrophils stimulated with fMLP the electrochemical treatment caused a 32% higher integral response as well as a 22% higher peak level compared to the neutrophils treated with chemical stimulant alone. The integrated oxidative burst response in the combined PMA and electric treatment was only 4.7% higher than in the cells treated with PMA alone, and no significant difference in the peak level was found. The results suggest that electric field treatment preferentially stimulates calcium-induced activation with ionomycin rather than calcium dependent activation with fMLP or PMA. PMID- 9353958 TI - Proton nuclear magnetic resonance detects leucine, 2,3-butanediol, and a prominent increase in the level of choline in the sera from patients chronically infected with schistosomiasis japonica. AB - 1H-NMR spectroscopy with 'Hahn' spin-echo pulse sequence has been employed to investigate the metabolic profiles of sera of chronic patients with schistosomiasis japonica, and compared with those of healthy volunteers and former patients who had been treated successfully. 1H-NMR clearly detected 2,3 butanediol and leucine, and markedly elevated levels of choline in sera from the chronic patients. Profiles of the sera from former patients were essentially similar to those from healthy volunteers, except that ketone bodies (3 hydroxybutyrate and acetone) were detectable in sera from 58% of the former patients but not in those of the normal controls patients. PMID- 9353959 TI - Determination of mercury levels in biological samples using the incomplete cubane type sulfur-bridged nitrilotriacetato molybdenum complex by a spectrophotometer. AB - Spectrophotometric determination of mercury levels in biological samples was investigated using incomplete cubane-type sulfur-bridged molybdenum complex, K2[Mo3S4(Hnta)3] 9H2O, ("NTA" complex; H3nta = nitrilotri acetic acid). The urine or organs of mice, which were either exposed to metallic mercury vapor or injected intraperitoneally with mercuric ion, were decomposed from four to twelve hours with a mixed solution of potassium permanganate and sulfuric acid. After the pretreatment, mercury in the urine and organs of mice was captured by the "NTA" complex. Absorbance of the resultant solution in the urine or organs of mice was also measured by a spectrophotometer under conditions similar to that of the exhalation. PMID- 9353960 TI - Production of peptide hormones and neurotransmitters by the immune system. PMID- 9353961 TI - Cytokines: influence on glial cell gene expression and function. PMID- 9353962 TI - Structural biology of cytokines, their receptors, and signaling complexes: implications for the immune and neuroendocrine circuit. PMID- 9353963 TI - Noradrenergic and peptidergic innervation of lymphoid organs. AB - It now is evident that extensive neural-immune anatomical connections exist between the nervous and immune systems, with close contacts of nerves with lymphocytes and macrophages. The presence of receptors for catecholamines and neuropeptides on these cells, coupled with functional evidence that these neural signals can modulate immune responses, brings these putative neurotransmitters to the forefront as a class of immunomodulatory molecules that can be investigated for possible benefit of disorders resulting from enhanced or suppressed activity of specific aspects of immune function. Furthermore, feedback from the immune system (cytokines) can act locally on lymphoid organ innervation to modulate transmitter release, and can act on the central nervous system via the vagus nerve to alter central pathways relevant to the immune system. It certainly is very clear that extensive bidirectional interactions occur between the nervous and immune systems, and that one system cannot be considered functionally without taking into account the state of activity of the other system. PMID- 9353964 TI - Neuroendocrine peptide receptors on cells of the immune system. PMID- 9353965 TI - Neuroendocrine peptide hormone regulation of immunity. PMID- 9353966 TI - Hormonal activities of cytokines. PMID- 9353967 TI - Interactive signaling pathways of the neuroendocrine-immune network. PMID- 9353969 TI - A case of nephrotic syndrome with rapid spontaneous remission in an elderly patient. AB - In July 1994, a 70-year-old woman was diagnosed as having nephrotic syndrome with proteinuria of 8 to 10 g/day and a serum albumin level of 1.8 g/dl. She was hospitalized in August 1994 for investigation. The urinary findings then normalized, with urinary protein and occult blood both negative and total urinary protein excretion at 0 g/day. A renal biopsy was performed, and spontaneous remission of minimal change nephrotic syndrome was diagnosed. This is an interesting case involving rapid remission of minimal change nephrotic syndrome in an elderly patient. PMID- 9353968 TI - Effects of acute arterial bleeding on renal sympathetic nerve activity in young and old anesthetized rats. AB - We compared the change in renal sympathetic nerve activity (RNA) during hemorrhage in young and old rats. Young (10 weeks) and old (80 weeks) male Wistar rats were anesthetized, and RNA together with arterial pressure (AP) and heart rate (HR) were measured under spontaneous respiration. Acute arterial bleeding, 1ml/100g body weight, was carried out to induce hypotension and hypovolemia. With bleeding, mean AP (MAP) decreased from 110 +/- 12 to 23 +/- 11mmHg (mean +/- SEM) and from 93 +/- 15 to 15 +/- 6mmHg in young and old rats, respectively. The difference in the change of MAP between the young and old rats was not significant. With bleeding, mean RNA increased by 46 +/- 21% and 20 +/- 16% in young and old rats, respectively. The increase in mean RNA of the old rats was significantly lower than that of the young rats. We estimated the gain of the baroceptor-renal sympathetic nervous system by using the formula delta RNA/RNA/delta MAP/MAP. The gain was 0.61 +/- 0.34 and 0.23 +/- 0.18 in young and old rats, respectively. The difference in the gain was statistically significant (p < 0.05). We concluded that the gain of baroceptor-sympathetic nerve system in acute arterial hemorrhage is attenuated with aging. PMID- 9353970 TI - Myeloperoxidase antineutrophil cytoplasmic autoantibody-associated glomerulonephritis in a very elderly patient with generalized vasculitis at autopsy. AB - An 86-year-old woman was admitted with hemoptysis and rapid deterioration of renal function. Renal biopsy disclosed necrotizing crescentic glomerulonephritis. Based on positivity for serum myeloperoxidase antineutrophil cytoplasmic autoantibody (MPO-ANCA), MPO-ANCA-associated glomerulonephritis was diagnosed. Steroid pulse therapy was performed, but the patient died after the second course. Autopsy revealed renal vasculitis with fibrinoid necrosis extending to the level of the arcuate arteries. Vasculitis was also observed in the liver, adrenal gland, uterus, and spleen, suggesting the presence of microscopic polyarteritis. This case demonstrates the broad spectrum of MPO-ANCA-positive vasculitis and suggests the need for a more effective therapy suitable for very elderly patients. PMID- 9353971 TI - A case of an interstitial tandem direct duplication of long arm of chromosome 4: 46, XY, dup (4) (q25q31.3) de novo. AB - We report a 4 2/12-year-old Japanese boy with a de novo direct tandem dup (4) (q25q31.3). The major clinical picture includes postnatal growth and psychomotor retardation, thick eye-lashes, a cleft lip, and large and prominent helix and antitragus. He did not have any hearing deficit. His eyegrounds were normal. There was no organ malformations including brain, kidney, liver, pancreas, gallbladder, urinary bladder, stomach, and heart. Routine hematological tests, blood chemistry including thyroid hormones, and urinalysis including urinary screening tests for congenital metabolic disorders showed normal results. He showed an electroencephalographic abnormality which could have resulted from mild aseptic meningitis at 2 months. Our case supports the idea that the association of thumb and renal deformities in duplication 4q syndrome is related to the region 4q22-q23 as many researchers have already pointed out. PMID- 9353972 TI - Distribution of sequence variation in the mtDNA control region of Native North Americans. AB - The distributions of mtDNA diversity within and/or among North American haplogroups, language groups, and tribes were used to characterize the process of tribalization that followed the colonization of the New World. Approximately 400 bp from the mtDNA control region of 1 Na-Dene and 33 Amerind individuals representing a wide variety of languages and geographic origins were sequenced. With the inclusion of data from previous studies, 225 native North American (284 bp) sequences representing 85 distinct mtDNA lineages were analyzed. Mean pairwise sequence differences between (and within) tribes and language groups were primarily due to differences in the distribution of three of the four major haplogroups that evolved before settlement of the New World. Pairwise sequence differences within each of these three haplogroups were more similar than previous studies based on restriction enzyme analysis have indicated. The mean of pairwise sequence differences between Amerind members of haplogroup A, the most common of the four haplogroups in North America, was only slightly higher than that for the Eskimo, providing no evidence of separate ancestry, but was about two-thirds higher than that for the Na-Dene. However, analysis of pairwise sequence divergence between only tribal-specific lineages, unweighted for sample size, suggests that random evolutionary processes have reduced sequence diversity within the Na-Dene and that members of all three language groups possess approximately equally diverse mtDNA lineages. Comparisons of diversity within and between specific ethnic groups with the largest sample size were also consistent with this outcome. These data are not consistent with the hypothesis that the New World was settled by more than a single migration. Because lineages tended not to cluster by tribe and because lineage sharing among linguistically unrelated groups was restricted to geographically proximate groups, the tribalization process probably did not occur soon after settlement of the New World, and/or considerable admixture has occurred among daughter populations. PMID- 9353973 TI - VNTR polymorphism in the Buenos Aires, Argentina, metropolitan population. AB - VNTR loci provide a wealth of information for human genetic research, ranging from gene mapping to paternity testing and forensic identification. In this study we report the construction, validation, and analysis of the first local genetic database for VNTR markers for Argentina. A sample of the metropolitan population of Buenos Aires was typed by means of six VNTR systems. Allele frequencies and expected heterozygosity were calculated. The sample set was further tested for departures from Hardy-Weinberg equilibrium and power of exclusion. Allele frequency distributions are compatible with previously reported data on Caucasian populations, and no departures from Hardy-Weinberg equilibrium were detected. PMID- 9353974 TI - Mutation rate and excess African heterozygosity. AB - Global studies of within-group genetic variation have revealed a tendency for some traits, but not all, to show higher heterozygosity in sub-Saharan African populations. Although excess African diversity has been interpreted as reflecting a greater "age" of sub-Saharan African populations, more recent research has shown that this excess is more likely a consequence of a larger African long-term effective population size. The observation that certain traits, particularly classic genetic markers and RFLPs, do not show this pattern has been interpreted as ascertainment bias. Here, I examine another possible factor: that excess African heterozygosity is in part a function of mutation rate. Simple equilibrium and nonequilibrium models of absolute excess heterozygosity are examined. The results indicate that there is little excess African heterozygosity for traits with low mutation rates and greater excess heterozygosity for traits with moderate to high aggregate mutation rates. Observed data are consistent with these models. Also, depending on population size and time depth, traits with high levels of mutation might show less excess heterozygosity than those with moderate to high mutation rates. Another measure of diversity, mean sequence divergence, shows an increase in excess diversity for traits with high mutation rates. PMID- 9353975 TI - Relationship of two apolipoprotein B polymorphisms with serum lipoprotein and lipid levels in African blacks. AB - Two DNA polymorphisms, insertion/deletion (ins/del) and XbaI, of the apolipoprotein B (APOB) gene and their relationships with serum lipoprotein and lipid levels were examined in two Nigerian populations from Benin (n = 828) and Sokoto (n = 394). Allele frequencies for both polymorphisms were comparable between the two populations, and genotype frequencies conformed to Hardy-Weinberg equilibrium expectations. Because lipid profiles are affected by many known sex differences, males and females were analyzed separately. Analysis of variance was performed to test equality of means of quantitative traits across genotypes; the dependent variables were first adjusted by stepwise linear regression for environmental covariates in both populations. Plasma apoB and HDL2 cholesterol levels varied significantly between ins/del genotypes in Benin males (p = 0.05) and Sokoto males (p = 0.02), respectively. No significant variation between XbaI genotypes was observed for any lipid trait in either population. Combining the ins/del and XbaI polymorphisms to form haplotypes revealed significant linkage disequilibrium in both the Benin (p < 0.0001) and Sokoto (p < 0.001) populations. Analysis of the two-site haplotype showed a significant effect on serum lipoprotein (a) levels in Benin females (p < 0.04) and on HDL2 cholesterol among Sokoto males (p = 0.01). The haplotype explained 4% of the adjusted phenotypic variation in lipoprotein (a) levels in Benin females and 8.5% of the adjusted phenotypic variation in HDL2 cholesterol levels in Sokoto males. These data indicate that the ins/del and XbaI polymorphisms in the APOB gene affect interindividual variation in serum lipoprotein and lipid levels in African populations. PMID- 9353976 TI - HLA class I and class II alleles and haplotypes in Mexican mestizos established from serological typing of 50 families. AB - We describe new information on the frequency and association of class II antigens (HLA-DR and HLA-DQ) of the major histocompatibility complex (MHC) in Mexicans. The study includes HLA-B typing and its association with the HLA-DR antigens determined in 50 families, which included 100 individuals. This family study allowed the establishment of the precise composition of the 200 HLA haplotypes, which cannot be obtained from unrelated individuals. The predominant antigens in decreasing order of frequency were B35, B39, and B61 at the B locus; DR4, DR5, and DR8 at the DR locus; and DQ3 at the DQ locus. The most common HLA-B,HLA-DR haplotype (considering broad specificities) was B16,DR4, with a frequency of 8.0%. Five HLA-B,HLA-DR haplotypes showed significant delta values (observed vs. expected frequencies) after correcting for the number of comparisons. On the other hand, the most common HLA-DR,HLA-DQ haplotypes were DR4,DQ3 and DR5,DQ3 with a frequency higher than 10%. Ten of the 17 HLA-DR,HLA-DQ haplotypes had significant postcorrection delta values. PMID- 9353978 TI - Ontogenetic changes in genetic regulation of fetal morphometrics in baboons (Papio hamadryas subspp.). AB - It is known that different genes are expressed during ontogeny; however, it is unclear how variation in that expression is associated with changes in growth patterns. The objective of this study is to assess how genetic variation in fetal morphology changes with ontogeny in baboons. Longitudinal measures of the head and femur (60 to 180 days gestation) were available for 892 pregnancies. We used a genetic model that allowed both the genetic and environmental variances (sigma 2G and sigma 2E) to change with age and estimated genetic and environmental correlations (rho G and rho E) between measurements at different ages. The results indicate a significant increase in the genetic variance for biparietal diameter and femur length but not for head circumference and fronto-occipital diameter. The rho G estimates for all measures decreased as the age between measures increased from 0 to 120 days, indicating that different groups of genes are expressed early in gestation and late in gestation. The rho E estimates dropped rapidly from 1 to 0 for all measures, indicating temporally localized environmental influences on fetal growth. Thus fetal morphometrics are significantly heritable and those genes that influence them show age-specific expression during ontogeny. PMID- 9353977 TI - Application of HLA class II polymorphism analysis to the study of the population structure of the Island of Krk, Croatia. AB - The population structure of the northern Adriatic island of Krk, Croatia, was studied using PCR methodology and nonradioactive oligonucleotide hybridization for the analysis of HLA-DRB1, DRB3, DRB4, DRB5, DQA1, and DQB1 polymorphisms. Allele frequencies, genetic kinship (R), and genetic distances (E2) were computed, and correlations between distance (genetic, linguistic, geographic) and kinship (migration) matrices were examined. The results, reflecting past (micro-) evolutionary processes, indicate that ethnohistorical and sociocultural events rather than geographic distances are the primary determinants of today's population structure of the island. PMID- 9353979 TI - Pattern and process of growth of the abnormal human fetus. AB - Identifying patterns of fetal growth alteration benefits both the clinician and the researcher. Twenty-four measurements in three variable sets (anthropometric measures, organ weights, and long-bone measures from radiographs) were taken on fetuses both with and without pathological conditions that are suspected to result in growth alteration. In addition, radiographs of each case were examined for the presence or absence of ossification centers. Based on least-squares regressions of the normal group, we calculated standardized residuals for the affected group to identify patterns of growth alteration. A large sample of fetuses between 15 and 42 weeks of gestational age with a variety of pathological conditions is described and evaluated for growth alterations. Symmetric and asymmetric growth alteration was detected in a small part of the sample and was predominantly isolated to fetuses in the late third trimester. Although patterns of growth alteration have been suggested as a means for noninvasive diagnoses of syndromes (such as trisomy 21), no consistent patterns are discernible in the current group. The sample provides a unique opportunity to evaluate fetal growth in terms of the interaction between genetic and environmental influences. PMID- 9353980 TI - Is length of life predictable? AB - A random sample of death records of adult males from the period 1967 to 1970 was chosen from the South Australian Registry of Births, Deaths, and Marriages. The natural parents of these individuals were identified by cross-referencing to birth certificates, and an extensive search was made of the death records for these parents. In this manner random families were selected for which, where possible, the cause of death and length of life of each family member were determined. Here, we analyze the association between sons and their parents in length of life and report the statistically useful correlations that were found. These correlations enable the calculation of a life table for a male conditional on his current age and the lifetimes of his parents. Comparisons are made with the uninformed population life table based solely on sex and year of birth. PMID- 9353981 TI - CTG repeat number in the nonaffected allele of myotonic dystrophy patients is not critical for disease expression. AB - To investigate whether unusual allele segregation might explain the dominant negative effect of the expanded allele for myotonic dystrophy on myotonin protein kinase mRNA metabolism, which is suggested to cause the disease, we determined the number of CTG repeats at the DM locus in the nonaffected alleles of 64 DM (dystrophia myotonia) patients. The relative distribution was then compared with the distributions obtained from alleles of the normal parents and normal siblings of DM patients. Comparison was also made with the allele distribution of normal subjects from the same geographic area. It appears that the CTG repeat number of the nonaffected allele in DM patients is not critical for the expression of the disease. PMID- 9353982 TI - Early recognition of postural disorders in multiple sclerosis through movement analysis: a modeling study. AB - In the present study, spontaneous postural behavior has been analyzed in freely standing multiple sclerosis (MS) patients, exhibiting no clinically assessable abnormalities of postural control. This population has been compared with two other groups, healthy people and hemiparetic patients. This latter group represents a situation where the central nervous system (CNS) lesion is precisely localized in one anatomical site and no signal-conduction disorders are present; i.e., it has an opposite anatomical character with respect to the MS at a preclinical stage. The hypothesis underlying the modeling study is the presence of a controller block working in a feedback posture control system. This controller block receives the body sway as input, and produces the corresponding ankle torque stabilizing the body, the latter being modeled as an inverted pendulum. The CNS damage, caused by MS, is supposed to be reflected in some detectable change in the structure of the controller of the posture control system. The identification of the controller has been performed by means of a parametric estimation procedure which employed as input sequences, data recorded by means of a movement-analysis (MA) system. Reported findings show a structural changes of the model of the controller block in the posture control system. This result may suggest the presence of an MS-specific reorganization of the posture control system. Some speculation is finally made on the black-box approach in comparison with traditional posturography, to arrive at hypothesizing a progression path for postural disorders. PMID- 9353983 TI - A hybrid computational model for ultrasound phased-array heating in presence of strongly scattering obstacles. AB - A computationally efficient hybrid ray-physical optics (HRPO) model is presented for the analysis and synthesis of multiple-focus ultrasound heating patterns through the human rib cage. In particular, a ray method is used to propagate the ultrasound fields from the source to the frontal plane of the rib cage. The physical-optics integration method is then employed to obtain the intensity pattern inside the rib cage. The solution of the matrix system is carried out by using the pseudo inverse technique to synthesize the desired heating pattern. The proposed technique guides the fields through the intercostal spacings between the solid ribs and, thus, minimal intensity levels are observed over the solid ribs. This simulation model allows for the design and optimization of large-aperture phased-array applicator systems for noninvasive ablative thermal surgery in the heart and liver through the rib cage. PMID- 9353984 TI - Electrical impedance tomography of complex conductivity distributions with noncircular boundary. AB - Electrical impedance tomography (EIT) uses low-frequency current and voltage measurements made on the boundary of a body to compute the conductivity distribution within the body. Since the permittivity distribution inside the body also contributes significantly to the measured voltages, the present reconstruction algorithm images complex conductivity distributions. A finite element model (FEM) is used to solve the forward problem, using a 6017-node mesh for a piecewise-linear potential distribution. The finite element solution using this mesh is compared with the analytical solution for a homogeneous field and a maximum error of 0.05% is observed in the voltage distribution. The boundary element method (BEM) is also used to generate the voltage data for inhomogeneous conductivity distributions inside regions with noncircular boundaries. An iterative reconstruction algorithm is described for approximating both the conductivity and permittivity distributions from this data. The results for an off-centered inhomogeneity showed a 35% improvement in contrast from that seen with only one iteration, for both the conductivity and the permittivity values. It is also shown that a significant improvement in images results from accurately modeling a noncircular boundary. Both static and difference images are distorted by assuming a circular boundary and the amount of distortion increases significantly as the boundary shape becomes more elliptical. For a homogeneous field in an elliptical body with axis ratio of 0.73, an image reconstructed assuming the boundary to be circular has an artifact at the center of the image with an error of 20%. This error increased to 37% when the axis ratio was 0.64. A reconstruction algorithm which used a mesh with the same axis ratio as the elliptical boundary reduced the error in the conductivity values to within 0.5% of the actual values. PMID- 9353985 TI - Application of sonomicrometry and multidimensional scaling to cardiac catheter tracking. AB - This paper describes a technique for tracking the three-dimensional (3-D) position of a cardiac catheter using sonomicrometry and the mathematical method of multidimensional scaling (MDS). Sonomicrometry is used to measure the distances between ultrasonic transceivers. MDS is then used to calculate the 3-D coordinates of the ultrasonic transceiver locations, including the catheter tip, from the measured distances. Feasibility of catheter tracking was initially studied using simulated data from a geometric model in which the actual coordinates of all transceivers were known. The method was then shown to be feasible in vivo by tracking a catheter-mounted piezoelectric transducer using seven reference crystals sewn to the epicardial surface of a sheep heart. Simulation results indicate that a catheter can be tracked with a root-mean square (rms) error of 1.51 +/- 0.05 mm and an average-distance error of e = 1.06 +/- 0.27 mm using 12 reference points. In vivo results showed acceptable stress values (G < 0.05) for 95% of the data samples with an average-distance error of e = 0.52 +/- 0.66 mm. These simulation and experimental results show that sonomicrometry and MDS can be used to accurately localize the 3-D position and track the motion of a catheter tip within the heart. PMID- 9353986 TI - Parametric representation and screening of knee joint vibroarthrographic signals. AB - We have been investigating analysis of knee joint vibration or vibroarthrographic (VAG) signals as a potential tool for noninvasive diagnosis and monitoring of cartilage pathology. In this paper, we present a comprehensive comparative study of different parametric representations of VAG signals. Dominant poles and cepstral coefficients were derived from autoregressive models of adaptively segmented VAG signals. Signal features and a few clinical features were used as feature vectors in pattern classification experiments based on logistic regression analysis and the leave-one-out method. The results using 51 normal and 39 abnormal signals indicated the superior performance of cepstral coefficients in VAG signal classification with an accuracy rate of 75.6%. With 51 normal and 20 abnormal signals limited to chondromalacia patella, cepstral coefficients again gave the highest accuracy rate of 85.9%. PMID- 9353987 TI - Spatio-temporal EEG source localization using simulated annealing. AB - The estimation of multiple dipole parameters in spatio-temporal source modeling (STSM) of electroencephalographic (EEG) data is a difficult nonlinear optimization problem due to multiple local minima in the cost function. A straightforward iterative optimization approach to such a problem is very susceptible to being trapped in a local minimum, thereby resulting in incorrect estimates of the dipole parameters. In this paper, we present and evaluate a more robust optimization approach based on the simulated annealing algorithm. The complexity of this approach for the STSM problem was reduced by separating the dipole parameters into linear (moment) and nonlinear (location) components. The effectiveness of the proposed method and its superiority over the traditional nonlinear simplex technique in escaping local minima were tested and demonstrated through computer simulations. The annealing algorithm and its implementation for multidipole estimation are also discussed. We found the simulated annealing approach to be 7-31% more effective than the simplex method at converging to the true global minimum for a number of different kinds of three-dipole problems simulated in this work. In addition, the computational cost of the proposed approach was only marginally higher than its simplex counterpart. The annealing method also yielded similar solutions irrespective of the initial guesses used. The proposed simulated annealing method is an attractive alternative to the simplex method that is currently more common in dipole estimation applications. PMID- 9353988 TI - Spectral decomposition in multichannel recordings based on multivariate parametric identification. AB - A method of spectral decomposition in multichannel recordings is proposed, which represents the results of multivariate (MV) parametric identification in terms of classification and quantification of different oscillating mechanisms. For this purpose, a class of MV dynamic adjustment (MDA) models in which a MV autoregressive (MAR) network of causal interactions is fed by uncorrelated autoregressive (AR) processes is defined. Poles relevant to the MAR network closed-loop interactions (cl-poles) and poles relevant to each AR input are disentangled and accordingly classified. The autospectrum of each channel can be divided into partial spectra each relevant to an input. Each partial spectrum is affected by the cl-poles and by the poles of the corresponding input; consequently, it is decomposed into the relevant components by means of the residual method. Therefore, different oscillating mechanisms, even at similar frequencies, are classified by different poles and quantified by the corresponding components. The structure of MDA models is quite flexible and can be adapted to various sets of available signals and a priori hypotheses about the existing interactions; a graphical layout is proposed that emphasizes the oscillation sources and the corresponding closed-loop interactions. Application examples relevant to cardiovascular variability are briefly illustrated. PMID- 9353989 TI - Estimation of conduction velocity distribution by regularized-least-squares method. AB - A novel technique for estimating the distribution of the conduction velocity of peripheral nerve fibers is described in this paper. In order to overcome the sensitivity of present methods to noisy data, a regularized-least-squares (RLS) method with a smoothing constraint and a self-adaptation of regularization parameter was adopted. The simulation results demonstrated that the improved technique maybe applied in clinical diagnosis because it yielded reliable and almost undistorted results even when the simulated data are severely contaminated by noise. PMID- 9353991 TI - Real-time frequency domain temperature and oxygen sensor with a single optical fiber. AB - The combined excited-state phosphorescence life-times of an alexandrite crystal and platinum tetraphenylporphyrin Pt(TPP) in a single-fiber sensor are used to monitor temperature and oxygen concentration in the physiological range from 15 45 degrees C and 0-50% O2 with precision of 0.24 degree C and 0.15% O2 and accuracy of 0.28 degree C and 0.2% O2. A 500-micron cubic alexandrite crystal bound to the distal end of a 750-micron-diameter optical fiber core and the Pt(TPP) coated circumferentially with a length of 1 cm from the end of the same fiber are excited with pulsed super-bright blue LED light. This apparatus uses a 125-kHz sampler for data acquisition and frequency domain methods for signal processing. The instrument amplifies both the dc and ac components of the photomultiplier output and band limits the signal to 20 kHz. The fundamental frequency of the excitation is set to 488.3 Hz and the highest harmonic used is the 35th. This bandlimited signal is sampled and averaged over a few hundred cycles in the time domain. The frequency domain representation of the data is obtained by employing fast Fourier transform algorithms. The phase delay and the modulation ratio of each sampled harmonic are then computed. At least four log spaced harmonic phases or modulations are averaged before decoding the two lifetimes of temperature and oxygen phosphorescent sensors. A component of zero lifetime is introduced to account for the excitation backscatter leakage through optical interference filters seen by the photodetector. Linear and second-order empirical polynomials are employed to compute the temperatures and oxygen concentrations from the inverse lifetimes. In the situation of constant oxygen concentration, the lifetime of Pt(TPP) changes with temperature but can be compensated using the measured temperature lifetime. The system drift is 0.24 degree C for the temperature measurement and 0.59% for the oxygen concentration measurement over 30 h of continuous operation. The instrumentation and methods allow for 6-s update times and 90-s full-response times. PMID- 9353990 TI - Automated feedback control of body temperature for small animal studies with MR microscopy. AB - A temperature control system consisting of a thermistor, signal processor, and computer algorithm was developed for magnetic resonance (MR) microscopy of small live animals. With control of body temperature within +/- 0.2 degree C of the set point, heart rate is stabilized and, in turn, repetition time (TR) during cardiac gated studies is less variable. Thus, image quality and resolution are improved. PMID- 9353992 TI - Nanoliter volume, high-resolution NMR microspectroscopy using a 60-micron planar microcoil. AB - Previous studies demonstrated the feasibility of using 100-microns inner diameter planar spiral inductors (microcoils) as detectors in 1H nuclear magnetic resonance (NMR) microspectroscopy. However, high-resolution NMR applications were not possible due to poor spectral resolution and low signal-to-noise ratio (SNR). These limitations in performance have now been largely overcome by using a nonconductive liquid fluorocarbon (FC-43) to minimize the effects of susceptibility mismatch between materials, and by carefully optimizing the microcoil geometry for maximum SNR. In this study, liquid samples were loaded into a fused silica capillary (75-microns inner diameter, 147-microns outer diameter). The capillary was positioned 50 microns above a 3.5-turn microcoil so that approximately 1 nL of the sample was present in the sensitive region of the microcoil. The microcoil was fabricated on a gallium arsenide substrate with an inner diameter of 60 microns, an outer diameter of 200 microns, trace width of 10 microns, trace spacing of 10 microns, and trace height of 3 microns. At 5.9 T (250 MHz) in 1H-NMR microspectroscopy experiments using a spectral width of 1 kHz, 4096 sampled data points, and a recovery delay of 1 s, a SNR of 25 (per acquisition) and a spectral linewidth of less than 2 Hz were obtained from a sample of water. These results demonstrate that planar microcoils can be used for high-resolution NMR microspectroscopy. Such coils may also be suitable for localized NMR studies at the cellular level and as detectors in capillary electrophoresis or microbore liquid chromatography. PMID- 9353993 TI - Electrothermal branding for embryo labeling. AB - A novel embryo labeling technique based on electrothermal branding is developed. Two types of micro branding irons are fabricated and tested. One utilizes 25 microns tungsten wire as the heating element. The other utilizes surface micromachining techniques to fabricate polysilicon branding irons. The thermal behavior of the branding irons and the heat distributions in the embryos are analytically modeled. Micron-scale labels on unfertilized bovine embryos are achieved. PMID- 9353994 TI - Factors affecting the accuracy of the boundary element method in the forward problem--I: Calculating surface potentials. AB - A comprehensive review of factors affecting the accuracy of the boundary element method (BEM) for calculating surface potentials is presented. A relative-error statistic is developed which is only sensitive to calculation errors that could affect the inverse solution for source position, and insensitive to errors that only affect the solution for source strength. The factors considered in this paper are: numerical approximations intrinsic to the BEM, such as constant potential versus linear-potential basis functions and sharp-edged versus smooth surfaced volumes; aspects of the volume conductor including the volume shape, density of surface elements, and element shape; source position and orientation; and effects of "refinements" in the numerical methods. The effects of these factors are considered in both smooth-shaped (spheres and spheroids) and sharp edged (cubes) volume conductors. This represents the first attempt to assess the effects of many of these factors pertaining to the numerical methods commonly used in fields such as electrocardiography (ECG) and electroencephalography (EEG). Strategies for obtaining the most accurate solutions are presented. PMID- 9353995 TI - Analog circuit for real-time computation of respiratory mechanical impedance in sleep studies. AB - The aim of this work was to develop a low-cost circuit for real-time analog computation of the respiratory mechanical impedance in sleep studies. The practical performance of the circuit was tested in six patients with obstructive sleep apnea. The impedance signal provided by the analog circuit was compared with the impedance calculated simultaneously with a conventional computerized system. We concluded that the low-cost analog circuit developed could be a useful tool for facilitating the real-time assessment of airway obstruction in routine sleep studies. PMID- 9353996 TI - Microelectrode arrays for electrophysiological monitoring of hippocampal organotypic slice cultures. AB - A three-dimensional platinum (Pt) microelectrode array embedded on a micromachined silicon (Si) substrate (porosity of 13%, via hole diameter of 40 microns) has been developed. Electrodes are 35-micron wide and 20-microns high, spaced 200 microns apart and arranged in an elliptic geometry. Integrated within a microperfusion chamber, the devices were used for stimulation and recording experiments of hippocampal slice cultures over a period of several days. PMID- 9353997 TI - HIV infection: legal aspects. PMID- 9353998 TI - Detection of anti-PPD IgG antibody and PPD-induced delayed type hypersensitivity in anterior uveitis patients. AB - Present study relates to the results of anti-PPD IgG, anti-A60 and antinuclear antibodies and PPD-induced delayed type hypersensitivity (DTH) in 17 anterior uveitis, (AU) patients. Results of anti-PPD IgG assay revealed detection of higher mean antibody level (O.D. 0.11 +/- 0.06) compared to healthy controls (O.D. 0.04 +/- 0.03), other eye disease controls (O.D. 0.05 +/- 0.003) and leprosy controls (O.D. 0.03 +/- 0.03). Anti-A60 IgM antibody assay revealed insignificant differences in mean antibody levels between various groups. Four of 17(23.5%) AU patients and 1(5.8%) subject each, belonging to other eye disease and healthy control groups had raised anti-nuclear antibody index. Findings of PPD skin test revealed detection of moderate to strong (2 to 4+) reactivity in 14 (82%). AU patients. Conversely, 13(76%) healthy controls and 8(47%) other eye disease controls gave mild (1+) reactivity. Results of this study suggested possible role of hypersensitivity to mycobacterial antigens in pathogenesis of anterior uveitis. PMID- 9353999 TI - Analysis of immunoglobulin deficiency cases: a five year study. AB - A total of 734 serum specimens from various clinical disorders along with 100 control samples from healthy subjects were processed for estimation of serum IgG, IgA and IgM employing single radial immunodiffusion procedure. Immunoglobulin deficiency, either selective or combined was noted in 31 males and 24 females in all age groups. Of the 55 cases encountered it was secondary immunoglobulin deficiency which was seen on a larger scale and encountered in patients with Multiple myeloma (16 out of 32) followed by Leprosy (14 out of 250), Lymphoma (5 out of 43), Malaria (4 out of 137), Burns (4 out of 52), Rheumatoid arthritis (2 out of 69) and non lymphoreticular malignancies (1 out of 41) in decreasing order of frequency. Primary immunoglobulin deficiency was observed in nine cases comprising of six belonging to Idiopathic late onset immunoglobulin deficiency, two of dysgammaglobulineamia and a solitary case of Ataxia telangiectasia. Panimmunoglobulin deficiency was observed in six cases, 11 had a dual deficiency while 38 showed deficiency of an isolated class with selective IgA deficiency in 20 cases. Furthermore, one patient each had total absence of IgG or IgA while IgM was not detectable in seven patients. A high suspicion index along with a regular rapport between the clinician and the laboratory personnel is necessary in the diagnostic set up of immunoglobulin deficiency states. PMID- 9354000 TI - Inflammatory myopathies--a clinicopathologic study. AB - In this study, clinical, histopathological and immunological profiles were analysed in ten patients with inflammatory myopathies. Polymyositis and dermatomyositis were more common than other forms of inflammatory myopathies. The pathogenetic mechanisms and distinguishing histopathological and immunological profiles between polymyositis and dermatomyositis have been highlighted. PMID- 9354001 TI - Significance of serum ferritin and lactate dehydrogenase in benign and malignant disease of breast. AB - Serum ferritin (SF) and lactate dehydrogenase (LDH) was estimated in 117 patients presenting with a breast lump and in 40 controls. Both pre and post treatment values were determined. Both the values were significantly higher in breast malignancies (p = 0.00) and also corresponded with the clinical stage and bulk of the tumour. The fall in post treatment values was proportional to the response to therapy. Persistent rise in values in the post treatment period was indicative of local recurrence of metastasis. PMID- 9354002 TI - Mucin histochemistry--a simple and effective method for diagnosing premalignant and early malignant lesions of lower gastrointestinal tract. AB - Mucin histochemistry of colo-rectal mucosa was studied in 124 biopsies which included 28 specimens from normal mucosa, 35 inflammatory lesions, 20 polyps, 26 malignant lesions and 15 specimens from transitional mucosa. The stains used were Haematoxylin and Eosin, Periodic acid schiffs, Alcian blue--Periodic acid schiffs (pH 2.5), Orcein--Alcian blue and Periodic acid sodium borohydride saponification. Normal mucosa, mucosa from inflammatory lesions, juvenile and hyperplastic polyps had neutral mucin in traces along with O-acylated sialomucins and sulphomucins. Predominant changes in adenomatous polyps and malignancy were a significant loss of sulphomucins and O-acylation at position C7, C8 or C9 in sialomucins. The changes in transitional mucosa were more on the pattern of malignant lesions. The alterations in the amount and/or pattern of mucin in colorectal mucosa has been found to predate frank morphological changes of malignancy. A routine use of mucin histochemistry in suspicious colorectal biopsies might be helpful in diagnosing premalignant and early malignant lesions in otherwise apparently normal colorectal mucosa or mucosa with chronic inflammatory changes and thereby reduce mortality. PMID- 9354003 TI - Sensitivity to the bactericidal effect of human serum of Proteus strains from clinical specimens. AB - A total of 257 Proteus strains isolated from urinary tract infection, blood, wound and faeces were studied. Of the strains tested 31 (12 percent) were serum sensitive, 182 (71 percent) were serum resistant and the remaining 44 (17 percent) showed intermediate sensitivity to the pooled normal human serum (PNHS). Strains isolated from adult urines and blood cultures were significantly more sensitive than strains of faecal origin (p < 0.01). No significant difference was seen between strains from faeces and wounds. PMID- 9354004 TI - Study of myopathies by histological and histochemical methods with special reference to staining for desmin expression. AB - An attempt was made to study the histological and histochemical changes as well as immunohistochemical changes in desmin expression occurring in four types of clinical myopathies e.g. Chronic ischaemic myopathy due to Buerger's disease (Group I), Carcinomatous myopathy (Group II), Metabolic myopathy (Group III) and Muscular dystrophy (Group IV). The number of cases studied were 16 cases, 15 cases, 4 cases and 5 cases respectively. The study revealed: (i) a combination of normal, degenerated, necrotic and regenerating fibres in different proportions in all the four groups having maximum number of degenerated fibres in Group I and Group IV, relatively more number of regenerating fibres in groups III and absence of necrotic fibres in Group I. (ii) Altered tinctorial property in most of the fibres indicating degenerated and regenerating fibres in all the groups with Masson's trichrome staining against inconstant staining with PTAH appear to be a good indicator for myopathy. (iii) The Desmin expression was week and irregular in most of the cases with most of the fibres probably due to reduction of desmin content probably indicating degenerated fibres, appear to be a good indicator for myopathy. (iv) Chronic ischaemic myopathy showed close resemblance with muscular dystrophy though no typical or distinct distinguishing feature could be identified in these four groups. PMID- 9354005 TI - Correlation of elevated levels of maternal serum beta-hCG in pregnancy induced hypertension and pregnancy outcomes in these patients. AB - This prospective randomized controlled study was conducted on 105 cases of pregnancy hypertension and 100 normotensive mothers in third trimester of pregnancy. The aim of the study was to determine the association between the elevated levels of maternal serum beta-hCG (MS-beta-hCG) with type and severity of hypertension and adverse pregnancy outcomes in these patients. Elevated levels were noted in 48.6% cases of pregnancy hypertension compared to controls, where the levels were elevated in 2% cases (P < 0.001). Elevated levels were more commonly seen in proteinuric pregnancy induced hypertension (32.4%) and also when the hypertension was severe (30.4%). The results of this study establish a strong association of elevated MS-beta-hCG levels with pregnancy hypertension and poor pregnancy outcomes in these patients. PMID- 9354006 TI - Study of association of Epstein Barr-virus with Hodgkin's disease. AB - Epstein Barr-Virus (EBV) is a lymphotropic herpes virus, well recognised for its oncogenic properties. In recent years substantial evidence has accumulated supporting a role for EBV in the pathogenesis of Hodgkin's disease. The epidemiologic and histologic features of Hodgkin's disease (HD) have long indicated a possible infective cause. Our study involves the detection of the EBV encoded latent membrane protein 1 (LMP-1) in 45 cases of Hodgkin's disease using immunohistochemical methods. In this study we detected LMP-1 positivity in the Reed-Sternberg and Hodgkin's cells in 31% of cases of HD. In mixed cellularity the positivity was 21.7% while nodular sclerosis exhibited positivity of 50% of the cases. The lymphocyte depletion subtype showed 100% positivity. All cases of lymphocyte predominance and the single unclassified case were negative for LMP. The demonstration of LMP-1 in Hodgkin's disease has important implications since it is one of the latent gene products which produces B Lymphocyte transformation. PMID- 9354007 TI - Comparative evaluation of breast lesions with the help of impression smears, histopathology and mammography. AB - A total of 148 surgically removed benign and malignant breast lesions were studied to correlate cytomorphologial features in impression smears and histopathological sections. Mammograms were taken prior to surgery. Role of mammography in detection of non-palpable breast lesions was enlightened. Impression smears helped in quick diagnosis (intraoperative) and overall accuracy obtained was 97.4%. Cluster predominant and Grade II nuclear grading on smear pattern was seen in 58.8% and 57.4% cases respectively amongst the 68 malignant cases. 45% cases revealed non-palpable breast lesions on mammography which was later confirmed on histopathology. PMID- 9354008 TI - Fish meal extract bile esculin agar (FMBE) a selective medium for Bacteroides fragilis group. AB - Fish meal extract bile esculin agar (FMBE) is prepared using Fish meal extract concentrate as the basal substance, for the selective isolation and presumptive identification of B.fragilis group. The efficiency of the medium was evaluated by growing stock cultures of B.fragilis groups as well as inoculating clinical specimens and comparing the results with Bacteroides bile esculin agar (BBE). All the 87 stock cultures of B.fragilis grew on FMBE and BBE. No other anaerobes tested grew on the medium. However 7 out of 65 neomycin resistant aerobes grew on the FMBE. From the 100 clinical samples, 62 strains of B. Fragilis group were recovered on FMBE and BBE, and 53 strains on supplemented BHIBA. The cost effectiveness, selectivity and the ability to detect esculin hydrolysis will enable FMBE as a suitable medium as comparable to that of BBE, if not superior. PMID- 9354009 TI - Characterization of mycobacterial species in clinically diagnosed cases of pulmonary tuberculosis and their HIV status. AB - A total of 75 clinically diagnosed and radiologically evident cases suggestive of pulmonary tuberculosis were selected for study. Sputum sample of each patient was screened for AFB by Ziehl Neelsen staining and culture. On examination 20 smears were found positive for AFB and 55 smears were negative by concentration method. A total of 23 samples were found to be culture positive and 52 were culture negative. Of these, 22 stains were identified as Mycobacterium tuberculosis, one was identified as M. Scrofulaceum. Of the 75 patients 3 were seropositive for HIV I antibodies. Out of these 3, one was found to be smear and culture positive and was identified as M. scrofulaceum. Other two seropositive patients were smear and culture negative for AFB. PMID- 9354011 TI - Inhibition of phagocytosis of E. coli by B. fragilis group--an in vitro study. AB - Bacteriodes fragilis group can inhibit the phagocytosis and killing of co existing aerobes. Forty six strains of B.fragilis group are tested for their ability to resist phagocytosis and inhibit the killing of indicator organism, E. coli by denoting the viable count. Among the species of B. fragilis group, the inhibitory index of the phagocytic system was the highest with B. fragilis followed by B. thetaiotamicron. None of the strains of B. fragilis group were phagocytosed or killed in this system. This property of resisting the phagocytosis and killing of E. coli by B. fragilis group might be the contributing factor towards their prevalence in mixed infections. PMID- 9354010 TI - Changing pattern of Vibrio cholerae serotype EL TOR and 0139 in Yavatmal (Maharashtra, India) during 1992 to 1994. AB - During 1992, 1993 and 1994 a total of 65, 123 and 142 faecal samples respectively yielded 9, 54 and 87 strains of V. cholerae. Simultaneous occurrence of EL TOR vibrio and non 01 (i.e. 0139) was noted during 1993 and 1994 with variations in relative and absolute prevalence of each serotype. Seasonal incidence is fairly consistent in this region. Both V. cholerae serotypes 01 and 0139 showed resistance to one or more drugs. PMID- 9354012 TI - A study on interrelationship of 60 HIV positive cases with coexistent oral candidosis and tuberculosis. AB - Sixty HIV seropositive cases studied of inter-relationship of coexistent infections with tuberculosis and oral candidosis. Presence of oral candidosis either symptomatic/asymptomatic as revealed on microbiological studies have been recorded and the significance discussed. Presence of oral candidosis apparent of nonapparent with heavy growth, yield on culture in HIV seropositive cases could be considered a pointer of conversion of HIV infection to a disease. This observation needs further evaluation. Relevant literature reviewed briefly. PMID- 9354013 TI - Gelatinous transformation of bone marrow. PMID- 9354014 TI - Aggressive angiomyxoma of the female perineum: a case report. AB - Clinical and pathological findings in a case of aggressive angiomyxoma, a distinctive soft tissue tumour arising in the vulva of a young female are described. The neoplasm is non malignant, but locally aggressive. PMID- 9354015 TI - Aggressive angiomyxoma. PMID- 9354016 TI - Mucinous adenocarcinoma of the renal pelvis--a case report. AB - A case of mucinous adenocarcinoma of the renal pelvis is added to those so far reported. Association with long standing chronic infection and large renal calculi is emphasized. The occult gross appearance of most of these tumours may lead the pathologist to overlook their presence. Hence, any kidney with the proper setting must be carefully looked for this entity. PMID- 9354017 TI - Fallopian tube carcinoma--a report of two cases. AB - Primary adenocarcinoma is a rare tumour involving the fallopian tube. Two such cases are reported; the first case was associated with papillary carcinoma of the paratubal cysts and the second occurred in a young female. PMID- 9354018 TI - Adult Wilms' tumour (report of two cases). AB - Wilms' tumour, an embryonic neoplasm, the most common renal tumour in childhood, had occasionally been reported in adults. Authors report two such cases and have reviewed the relevant literature. While Wilm's tumour in children classically demonstrates the curative potential of combined modality treatment, no such clear guide lines are available for those occurring during adulthood. Pathologic diagnosis of adult Wilm's tumour is difficult because of the multiplicity of undifferentiated adult tumours that must be considered in the differential diagnosis. As no predictive parameters for optimal therapy exist, a combination of surgery, irradiation and chemotherapy probably is indicated for all stages of disease. PMID- 9354019 TI - Bilateral renal angiomyolipoma in association with tuberous sclerosis. AB - Renal angiomyolipoma with tuberous sclerosis has not yet been reported in Indian literature. We report a case of bilateral angiomyolipoma associated with tuberous sclerosis in a 25 year old man. PMID- 9354020 TI - Metastatic carcinoma in uterine leiomyoma. AB - Metastasis of tumours are known to occur in the rarest of sites in the human body. But one of the truly rare phenomena in surgical pathology is to find the metastasis of one tumour into another. We report one such case of metastatic carcinoma in uterine leiomyoma from an occult primary site. PMID- 9354021 TI - Multiple myeloma complicating HIV-infection. A case report. AB - A case of multiple myeloma associated with HIV disease and hepatosplenomegaly presented to us as pyrexia of unknown origin, is reported. Because of paucity of such cases in the literature, the case is dealt in detail and the literature reviewed briefly. PMID- 9354022 TI - Haematological manifestation of HIV infection. PMID- 9354023 TI - Evaluation of coagglutination technique for detection of rheumatoid factor. PMID- 9354024 TI - Problems in interpretation of results of HIV tests. PMID- 9354025 TI - Analysis of dental health education activity carried out by dentistry students. AB - During their career, students of dentistry acquire techniques which they will employ later on their patients. Since the health of these patients and society itself cannot be left exclusively to the professional activity of these future dentist, an experimental extra-class experience was carried out with pupils from primary and secondary schools. The students gave the pupils presentations on Nutrition, Calcium and Phosphorus and Caries. The aim of this study was to evaluate: a) the influence of society on the teaching process, and training during the first university year, and b) epidemiologic aspects. Our results indicated that 32.2% of the primary and 15.6 % of the secondary school pupils were unfamiliar with the disciplines presented by the students. According to the evaluations 78.1% of the primary and 94.0% of the secondary school pupils achieved or exceeded the minimum requirements considered necessary for correct nutritional and oral health habits. Evaluations of the students demonstrated that the students who had participated in the activity did not have knowledge superior to that of non-participating students. Therefore this experience can be considered an exercise in utilizing human resources for primary prevention rather than an innovative methodology which improves the teaching-learning process. It also seems recommendable in view of the low cost of the exercise. PMID- 9354026 TI - Flouride release from various restorative materials. AB - Fluoride release from six light-activated restorative materials, including two resinmodified glass-ionomers, two composites, and two compomers, was evaluated and compared with one conventional acid-based glass-ionomer cement. The amount and rate of release varied among the tested materials. Both resin-modified glass ionomers and the conventional acid-base glass-ionomer cements released more fluoride then the composites and compomers (p < 0.05). Additionally, composite materials released less fluoride than compomer materials (p < 0.05). Release of fluoride by the tested materials showed a significant decrease after all the tested time intervals. PMID- 9354027 TI - Intraoral minor salivary gland tumors: a retrospective study of 129 cases. AB - From 1970 to 1996, 129 cases of intraoral minor salivary gland tumors were diagnosed at the Department of Pathology, Nihon University School of Dentistry. The diagnosis of each case was based on the 1991 WHO classification. Eighty benign and 49 malignant minor salivary gland tumors were found in the approximately 9,300 oral biopsies submitted during the 27-year period. Pleomorphic adenomas were the most commonly histologic type of the benign tumors identified and 51% of the malignant tumors were diagnosed as mucoepidermoid carcinoma. The most common primary location of the tumors was the palate. Sixty percent of all tumors occurred in females and the peak age for incidences of all tumors was found in the third, fourth, sixth and seventh decades. These results were compared with those of the studies in different world population groups. PMID- 9354028 TI - Solubilizing efficiency of different gutta-percha solvents: a comparative study. AB - A study was conducted to comparatively evaluate the efficiency of different solvents for dissolving gutta-percha. Halothane, chloroform, xylene, acetone, isopropyl alcohol, turpentine, oil of mela-leuca and eucalyptol were used as solvents for dissolving standardized gutta-percha discs. Halothane, chloroform and xylene were markedly superior solvents of gutta-percha in comparison with the others. There was no significant difference among the three (p > 0.05). Eucalyptol, turpentine and oil of melaleuca were relatively less efficient. Acetone and isopropyl alcohol did not dissolve gutta-percha, being similar in this respect to distilled water. PMID- 9354029 TI - Anti-bacterial action of onion (Allium cepa L.) extracts against oral pathogenic bacteria. AB - In this study, the focus was on the antibacterial activity of onions. This study researched the activities of onion extracts on Streptococcus mutans and Streptococcus sobrinus, the main causal bacteria for dental caries, and Porphyromonas gingivalis and Prevotella intermedia, considered to be the main causal bacteria of adult periodontitis. The results showed that the onion extracts possess an effect on all test bacterial strains (S.mutans JC-2, S. sobrinus OMZ176, P. gingivalis ATCC 33277 and P. intermedia ATCC 25611), and the effects were bactericidal against cultured and resting bacterial cells. The activity of the onion extracts was stable even after 48 hours in the culture medium. This result suggests that no decomposition or volatility of onion extracts occurred in the culture medium. The antibacterial activity of onion extracts was not markedly influenced by cysteine (10 mM) treatment. However, activity significantly decreased with alkali treatment. Grated onion left to stand at 37 degrees C for 48 hours did not show antibacterial activity. Also, activity of steam treated (100 degrees C, 10 min.) onion was not observed. Using avicel plate by thin layer chromatography with the solvent of n-butanol:acetic acid:water (3:3:1), the main component of the substance (the substance which develops color with ninhydrin) was observed at an Rf value of about 0.9. PMID- 9354030 TI - Effects of YM-14673, a thyrotropin-releasing hormone analogue, injected into the shell and the core of the nucleus accumbens on production of repetitive jaw movements in rats: comparison with the effects of a dopamine D1 and D2 receptor agonist combination. AB - The present study examined whether the shell and the core of the nucleus accumbens play a differential role in the display of YM-14673-induced jaw movements in rats. For that purpose the effects of YM-14673 were compared to those of a SKF 82958 and quinpirole combination, a dopamine D1 and a D2 receptor agonist respectively, that is known to functionally differentiate these two subregions of the nucleus. Consistent with the previous report, bilateral injections of a mixture of SKF 82958 (5 micrograms) and quinpirole (10 micrograms) into the shell of the nucleus accumbens produced repetitive jaw movements, whereas similar injections of the mixture into the core did not induce such an effect. In contrast, there was no regional difference in the effects of YM-14673 on the production of repetitive jaw movements. Thus, both bilateral injections of YM-14673 (0.1 or 1.0 microgram) into the shell or the core produced similar repetitive jaw movements in a dose-related manner. Moreover, the pattern of oral movements induced by YM-14673 differed from that induced by the mixture of SKF 82958 and quinpirole; frequent tongue protrusions were evident in rats treated with the mixture but were not seen in YM-14673-treated rats. It therefore appears that, unlike the effects of the mixture of dopamine D1 and D2 receptor agonists, the effects of YM-14673 in the shell on the production of rat jaw movements do not differ from the effects of the compound in the core. PMID- 9354032 TI - Arg-Gly-Asp(RGD) peptides inhibit Streptococcus mitis to adhere to fibronectin. AB - Fibronectin (Fn) is a multifunctional adhesive protein found on cell surfaces as well as in plasma. It is also believed to play an important role in bacterial adherence to host tissues. Molecular analyses of Fn have shown that the amino acid triplet arginine-glycine-aspartic acid (RGD) sequence functions as a binding site. We examined the role of the RGD sequence on bacterial adherence to Fn. The pretreatment of Streptococcus mitis with synthetic RGD-containing peptide reduced the number of bound bacteria to the Fn coated plates by 76%. In contrast, a control peptide containing the RGE sequence showed no inhibition. These data indicate that synthetic RGD peptides may be useful for the inhibition of bacterial adherence to Fn on host cell surfaces. PMID- 9354031 TI - Epithelial hyperplasia of the extra-glandular excretory ducts of human minor salivary glands: a histopathologic study. AB - The histologic features of normal and hyperplastic epithelia of the extra glandular excretory ducts of human minor salivary glands were studied, and their pathologic significance evaluated. Normal duct epithelium consisted of two layers: inner columnar cells, and basal cubical or squamous cells. A few goblet cells were present among the inner cells. Hyperplasia of the duct epithelia occurred focally or entirely, and was classified into the following histologic types: (1) simple hyperplasia, and (2) metaplastic hyperplasia, which were divided into (a) mucous cell hyperplasia, (b) oncocytic hyperplasia and (c) squamous cell hyperplasia. Squamous cell hyperplasia was subdivided into (i) acanthotic type and (ii) reserve cell-like type with or without dysplasia. Simple or metaplastic epithelial hyperplasia of the extra-glandular excretory ducts of minor salivary glands may be induced by chronic inflammation or other types of irritation, and proliferating cells of such regenerating tissue sometimes exhibit features reminiscent of a neoplastic process. Furthermore, it is suggested that metaplastic epithelial hyperplasia of the excretory minor salivary gland ducts could be the site of origin of tumor development, i.e., some oral squamous cell carcinomas may arise from primary lesions in the hyperplastic epithelium of the extraglandular excretory minor salivary gland ducts. PMID- 9354034 TI - Parting ways at the bridge--when our accompaniment can go no further. PMID- 9354033 TI - Immunohistochemical and biochemical characterization of sulphated proteoglycans in embryonic chick bone. AB - The type and distribution of sulphated proteoglycans (PGs) in the midshaft subperiosteal bone of 15-18-day embryonic chick femurs were studied immunocytochemically and biochemically, using four monoclonal antibodies (MAb 2B6, 3B3, 1B5, and 5D4). These MAb specifically recognize epitopes in chondroitin 4-sulphate (C4-S) and dermatan sulphate (DS); chondroitin 6-sulphate (C6-S) and unsulphated chondroitin (C0-S); C0-S; and keratan sulphate (KS) respectively. Immunohistochemistry showed that staining of C4-S, DS, and KS, but not of C6-S and C0-S, was limited to osteoid, the cell surface of osteocytes, and to the walls of osteocytic lacunae and bone canaliculi in 15-18-day embryonic specimens. However, no significant difference in the distribution and intensity of immunostaining was observed in these specimens. Bone proteins were extracted from fresh 18-day embryonic specimens with a three extraction procedure, 4 M guanidine HCl (GdnCl, G-1 extract), 0.4 M EDTA (E-extract), followed by GdnCl (G-2 extract), to characterize mineral binding and collagenous matrix associated PGs in E- and G2-extracts respectively. Western blot analysis of E- and G2-extracts demonstrated that chondroitinase ABC-digested PGs with a molecular weight (Mr) approximately of 45,000 containing GAGs predominantly corresponding to C4-S and/or DS, with no detectable C6-S or C0-S present in the mineral and matrix phase, whereas KSPGs having an Mr of approximately 72,000 are only present in the mineral phase. These results indicate that embryonic chick bone contains small PGs having C4-S, DS, and KS chains with preferential localization to osteoid, the cell surface of osteocytes, and to the walls of osteocytic lacunae and bone canaliculi. PMID- 9354035 TI - Evaluation of an interdisciplinary training program in palliative care: addressing the needs of rural and northern communities. AB - Our study was a pilot test of an interdisciplinary training program in palliative care to improve the quality of care to terminally ill cancer and AIDS patients in rural and northern communities in Manitoba. The program involved two weeks of intense palliative care training for nurses, social workers, physicians, and volunteers. Four teams were trained during a six-month period. A repeated measures design was used to assess the effectiveness of the program. Results indicated that health professionals' knowledge about care of the dying, care of individuals with HIV/AIDS, and attitudes toward care of the dying improved upon completion of the training program and remained improved three months following the program. Improvements in use of medications, increased attention to family care, increased discussion of DNR orders, and increased consultation related to symptom management were evident following the training program. The parallel training program for volunteers was also judged to be effective. PMID- 9354036 TI - Provisional educational needs of health care providers in palliative care in three nursing homes in Ontario. AB - Health care providers in three nursing homes in Ontario were surveyed to determine educational needs, barriers to meeting these needs, and the preferred format for education. Of the 415 health care providers asked to participate, 225 completed the questionnaire. Need was expressed for the majority of the 35 educational topics identified, including the role of the palliative care team, management of physical symptoms, pharmacological and non-pharmacological management of pain, stress management, spiritual needs, culture and death, and counseling. Group discussions and seminars were favored over traditional lectures. The primary factors influencing attendance at a palliative care workshop were loss of pay and time and location of the workshop. PMID- 9354037 TI - Pain and the choice to hasten death in patients with painful metastatic cancer. AB - Unrelieved pain has been cited as an important reason why cancer patients may seek to hasten their deaths. We interviewed 48 patients with painful metastatic cancer to ascertain their interest in various active and passive modes of hastening death. Ninety percent of these patients supported the general right of terminally ill patients to passive modes of hastening death and 80% supported the right to active modes such as assisted suicide and euthanasia. If they developed severe pain that could not be relieved, 80% would instruct their physician write a "do not attempt resuscitation" order, 40%-50% would want to receive suicide information or a lethal prescription from their physician, and 34% would request a lethal injection from their physician. Current pain and depression levels were not associated with interest in hastening death, but current somatic symptom burden was significantly associated with this interest. PMID- 9354038 TI - Communicating with patients with advanced cancer. AB - We conducted a prospective study of 130 unselected adults admitted to hospital with advanced malignancy. The study assessed how patients perceived information conveyed to them by physicians and the level of communication between patients and health care staff on questions relating to patients' understanding, pain control, and sense of well-being. Nearly 10% of patients were unaware of a diagnosis of cancer. Of those who knew their diagnosis, one quarter stated that the diagnosis was not disclosed in a clear or caring manner. One third of patients had an incomplete understanding of their prognosis, and patients generally overestimated their understanding when compared to hospital medical officers (HMOs) and nurses. The severity of pain was underestimated by the HMOs in 63% to 89% of patients. Thus HMOs require specific training in communication skills and the management of chronic cancer-related pain in patients with incurable cancer. Professional interpreters should be employed for all patients who are not fluent in English. PMID- 9354040 TI - Increasing pain and escalating opioid doses in a patient with cancer. PMID- 9354039 TI - Evaluating a home palliative care service: development of indicators for a continuous quality improvement program. PMID- 9354041 TI - Nebulized opioids in the treatment of dyspnea. PMID- 9354043 TI - Human rights and women's health. PMID- 9354042 TI - Patients with troublesome sweating. PMID- 9354044 TI - Human rights and the politics of risk and blame: lessons from the international reproductive health movement. AB - Recent debates about the "politicization" of public health obscure the ways in which epidemiological concepts of risk are routinely used in the legal and political systems to apportion blame and responsibility for poor health. This article uses the example of reproductive health and rights to argue that new understandings of the connection between socioeconomic conditions and poor health will only generate change when they are reframed into political claims and pressed by social movements. In this connection, human rights language, principles, and practice hold great potential for the US reproductive rights movement, which has sometimes been constrained by the narrow scope of court rulings. PMID- 9354045 TI - Transformative combinations: women's health and human rights. AB - From the human rights perspective proposed in this article, a woman's good or ill health reflects more than biology or individual behaviors; it reflects her enjoyment (or lack thereof) of fundamental human rights that enable her to exercise basic power over the course and quality of her life. The "structural" view of health that such a human rights perspective suggests is concerned first with identifying the effects of social, economic, and political relations on women's health and then with promoting "interventions" aimed at transforming the laws, institutions, and structures that deny women's rights and well-being. Yet, traditional human rights law and practice have been limited to narrowly defined abuses by public officials against individuals that fail to capture the most pervasive denials of women's rights, which, though rooted in systematic discrimination, are frequently played out in so-called "private" institutions, primarily within the family. The experiences of women's health advocates in addressing complex women's health issues makes it clear that women's lack of access to economic and political power in the public sphere creates the conditions under which they are discriminated against and physically and sexually abused in the private sphere. Combining the pragmatic understanding of women's health professionals with an expansive conception of human rights norms has the potential to transform the fields of women's health and human rights. PMID- 9354046 TI - Facts and fiction regarding female circumcision/female genital mutilation: a pilot study in New York City. AB - Little information on the practice of female circumcision/female genital mutilation (FC/FGM) in the West is currently available. Recent legislative efforts have largely ignored the main public health issue: the needs of girls and women living with circumcision in a new country that condemns the practice and where health care providers are not trained in the management of its complications. We report here on a needs assessment designed to determine the extent of FC/FGM in African immigrant communities in New York City, the health and social service needs of African immigrant women, and the training and information needs of their providers. Obstetrics/ gynecology providers in 8 of New York's 11 public hospitals and 10 maternal infant care/family planning (MIC/FP) clinics were surveyed, along with 20 women from FGM-practicing countries. Quality services for women living with circumcision can be fostered if care is provided in a sensitive and culturally appropriate manner, with thorough training and education of health care providers on the physical and mental health consequences and clinical management of FC/FGM, along with counseling guidelines, interdepartmental linkages, referrals and integrated service delivery, and the provision of translators and information in African languages. PMID- 9354047 TI - Female circumcision/female genital mutilation in the United States: legislation and its implications for health providers. AB - Criminal laws prohibiting female circumcision/female genital mutilation (FC/FGM) have recently been passed in the US Congress and in several state legislatures. The full effect of criminalization on prevention and on the overall well-being of immigrant groups from FC/FGM-practicing countries is currently unknown and will ultimately depend on, among other things, the precise interpretation of the laws by courts and local authorities. Meanwhile, the content of these laws prompt questions about their intended and inadvertent effects on FC/FGM, including: What acts are criminalized under these laws? Will criminalization prevent them? Do the laws have the potential to do more harm than good? The appropriateness of prosecuting FC/ FGM under existing child protection statutes is also raised. PMID- 9354049 TI - Medical and psychological examination of women seeking asylum: documentation of human rights abuses. AB - Human rights abuses of women are ubiquitous throughout the world. Those perpetrated by governments entitle women to seek political asylum, and many women refugees do so in the United States. The asylum process often requires medical or psychological evaluations to corroborate women's reports of torture or other abuses. This article provides an overview of how to conduct such examinations and how to document findings for the asylum process. PMID- 9354048 TI - The impact of economic embargoes on the health of women and children. AB - Economic embargoes have become common since the end of the Cold War. This review of health changes in Iraq, Haiti, Yugoslavia, Nicaragua, South Africa, and Cuba indicates that the major impact of decreased access to health goods occurs among the most politically and economically vulnerable sectors of the population, particularly women and children under five years of age. While some societies are skilled at attenuating the negative health effects of economic embargoes, nearly all countries have demonstrable deterioration in the health sector, despite major attempts to provide humanitarian assistance. Recommendations for reducing the negative impact of economic embargoes on civilian populations include exempting humanitarian goods (food and medical supplies) and international agreements to define exempt goods and guarantee their delivery. PMID- 9354050 TI - Political rape as persecution: a legal perspective. AB - The use of rape as a tool of persecution is not new, but recognition of the political rape of women as a violation of internationally protected human rights and as a basis for political asylum is. Over the last several years, a number of advances have been made, including human rights instruments that recognize the need to protect women from rape and other sexual abuse; guidelines from the United Nations High Commission for Refugees and several countries that acknowledge the political nature of rape and the difficulties experienced by women attempting to assert claims to asylum based on rape or other sexual abuse; and a number of important decisions by individual governments to provide protection to survivors of rape. Legal advancements for women in this area have depended largely on the assistance of medical and psychological experts who have been able to educate adjudicators and advocates on the effects of sexual harm, provide expert testimony in individual asylum cases, and provide critical treatment and support to survivors as they work their way through the process of obtaining legal protection. PMID- 9354051 TI - Profiles of four women. Health and human rights activists. AB - This article briefly profiles four women physicians working for health and human rights around the world. Dr. Ruchama Marton, an Israeli psychiatrist and activist for peace in the Middle East, is a founder of Physicians for Human Rights/Israel. Dr. Jane Green Schaller is a US pediatrician whose 1985 trip to South Africa initiated her human rights involvement, which includes the founding of Physicians for Human Rights. Dr. Judith van Heerden, a primary care physician in South Africa, has worked for reform of prison health care, to establish hospice care, and, most recently, for acquired immune deficiency syndrome (AIDS) education for medical students. Dr. Ma Thida, the only physician not interviewed for this article, is currently held in a Burmese prison because of her work on behalf of the National League for Democracy. The profiles suggest the breadth of human rights work worldwide and are a testament to what physicians can do. PMID- 9354052 TI - The Tuskegee Syphilis Study and women's health. AB - In May 1997, President Bill Clinton apologized for the Tuskegee Syphilis Study. The President's action underscores that in the 25 years since its public revelation, the study has moved from a singular historical event to a powerful metaphor that symbolizes racism in medicine, misconduct in human research, the arrogance of physicians, and government abuse of black people. The Tuskegee Syphilis Study also has implications for women's health. Discussion of the study usually ignores its impact on the wives of the victims. In addition, African American women may be more reluctant to participate in clinical trials because of the shadow cast by the syphilis study and other incidents of medical abuse. Finally, the Tuskegee Syphilis Study reminds us that the battle against racism must be an integral part of the campaign to improve women's health. PMID- 9354053 TI - "A whiter shade of pale". PMID- 9354054 TI - Cost effectiveness in optometric health care. PMID- 9354055 TI - Albinism: an update and review of the literature. AB - BACKGROUND: Albinism can be a diagnostic challenge to the optometrist, with ocular albinism the entity most likely to be overlooked or misdiagnosed. Albinism should be suspect in a child with nystagmus. METHODS: Albinism is best diagnosed by electron microscopy of skin or hair bulbs. Measuring the flash visually evoked response (VER) is the best way to confirm the abnormal decussation of the optic nerve fibers. Transillumination of the iris can be performed to see if it lacks pigment on its posterior surface. Optometrists should also be alert for nystagmus, strabismus, lack of stereopsis, and poor fusional capacity. Most people with albinism have photophobia and reduced acuity, and many have defective hearing. RESULTS: This review describes the various kinds of albinism and summarizes associated ocular manifestations with pertinent forms of the disorder. CONCLUSIONS: Optometrists are responsible for detection of albinism and provision of optical aids and related advice to afflicted patients. These patients also need appropriate counselling and genetic studies of family members. PMID- 9354056 TI - The relationship between moderate hyperopia and academic achievement: how much plus is enough? AB - BACKGROUND: There is evidence linking uncorrected hyperopia in children with academic learning problems. METHODS: This study was designed to test that hypothesis and--given supportive data--to then address a second topic: the minimal amount of uncorrected hyperopia that appears to impede elementary school performance. RESULTS: The refractive status and achievement test scores of 782 first-through-fifth grade children were compared. CONCLUSIONS: Statistical analysis indicated significantly lower achievement test scores among hyperopic children whose refractive errors exceeded 1.25 D (ANOVA F = 12.51; df = 4; p = 0.014). PMID- 9354057 TI - New England eye care survey. AB - BACKGROUND: Descriptive statistics on patient or consumer use of eye care are important for health care policy and professional development. Most survey research has been through doctors or doctors' offices. Some consumer surveys have been completed by direct mail, but information from direct mail surveys suffers from respondent self selection. This study joins a growing number of surveys designs that use random-digit telephone survey methods, which have less self selection bias. METHODS: This study describes patient use and perceptions of eye care services in three New England areas using random-digit telephone survey. Nine hundred eighteen calls were attempted. A standardized interview and survey questionnaire was used for each phone contact. RESULTS: Of 163 respondents (18% response rate), it was found that most patients were satisfied with the care, time, discussion, and fees provided by their eye care practitioner. Most (78%) planned on returning to the same practitioner in the future. Sixty percent of participants saw an optometrist, 29% saw an ophthalmologist, and 11% were unsure. In a subsample, differences in perception between optometric and ophthalmologic care was investigated, and no statistically significant differences (p < 0.05) were found. CONCLUSIONS: The study findings are similar to others in terms of market share between optometric and ophthalmologic settings for primary eye care. Several new findings were measured and discussed, and the data may be helpful for policy decisions. PMID- 9354058 TI - Intraocular pressure in 528 university students: effect of refractive error. AB - BACKGROUND: Very few studies measuring the intraocular pressure (IOP) in students are available. Recognizing the higher prevalence of myopia among students, IOP was analyzed in 528 university students according to age, gender, and refractive error. METHODS: The IOP was measured in 1,056 eyes with the Keeler Pulsair noncontact tonometer. The refractive error was determined with an autorefractometer (Shin-Nippon brand, model QR-007). RESULTS: The analysis of variance (ANOVA) showed no significant influence of age and gender on students' IOP (mean IOP, 15.77 +/- 2.67 mmHg). Refractive error did not significantly influence the mean of IOP (ANOVA) in the different categories: (a) severe myopia, (b) medium myopia, (c) emmetropia and mild ametropia, and (d) hypermetropia. The chi 2 test revealed significant differences, as a result of age, between the eyes with IOP < 21 mmHg or > or = 21 mmHg (6.3%). Nevertheless, there was no significant IOP difference as a result of gender or refractive error among eyes with normal IOPs and eyes with IOP above 20 mmHg. CONCLUSIONS: Among university students, intraocular pressure is not influenced by age, gender, or refractive status. Likewise, except for age, there is no significant correlation between any of the other variables and the normal range of IOP. After the age of 25 years, there is a slight increase in the number of eyes with an IOP above 20 mmHg. PMID- 9354059 TI - Combination ptosis crutch and moisture chamber for management of progressive external ophthalmoplegia. AB - BACKGROUND: A 46-year-old woman with a long-standing history of chronic progressive external ophthalmoplegia manifested a primary visual symptom of bilateral ptosis. Previous lid surgery was unsuccessful because of severe sight threatening exposure keratitis. CASE REPORT: The patient's problem was managed by fabricating a spectacle-mounted combination ptosis crutch and moisture chamber. The custom device successfully provided both cosmetic and visual relief for the patient while maintaining corneal integrity. CONCLUSIONS: This case demonstrates the effectiveness of a dual-purpose appliance that offers a cosmetically viable alternative to ptosis patients who are high-risk surgical candidates because of potential corneal dehydration. PMID- 9354060 TI - Training initiative list scheme (TILS) for minimal access therapy: the MATTUS experience. AB - The objective of the MATTUS intercollegiate exercise was to set up and audit a training initiative list scheme (TILS) by which funds are awarded to Trust hospitals for operative sessions used specifically for the training of staff in minimal access therapy (MAT). A prospective centralized audit of TILS involving nine Trust hospitals in Scotland over a 12-month period (1 March 1995-end of February 1996) was carried out. These hospitals had contracted for 510 4-h training sessions (389 for minimal access surgery, 121 for allied interventional techniques) by MATTUS accredited consultant tutors. The scheme covered training in technical competence for Minimal Access Surgery (MAS), interventional flexible endoscopy and interventional radiology within Scottish Hospitals. The main outcome measures used in the audit were trainee completion rates, conversion rates, morbidity and mortality, assessment of training received by trainees and assessment of aptitude by the trainers. The results were as follows. Of 510 sessions, 482 (95%) were completed within the deadline. Of these, 463 sessions were audited (367 for MAS, 69 for flexible endoscopy and 27 for interventional radiology). During these sessions, 817 operations/procedures were performed (781 training and 36 developmental). A total of 544 operations were performed during 339 MAS training sessions and 237 radiological/flexible endoscopy procedures in 96 MAT training sessions. The trainee was the principal operator in 643 (82%) procedures and completed the task in 581 (74%) cases. Four per cent of the MAS operations (22/544) required conversion. Post-operative complications occurred in 42 out of 817 patients (5%). Four patients, all with advanced malignancy, died within 30 days of the procedure. Trainees graded 355 sessions as excellent, 109 good, two as average and one as unsatisfactory. The tutors graded their trainees' aptitude to perform the operation as excellent in 34%, good in 53%, average in 11% and poor in < 1%. The training initiative list scheme which allows unhurried training in MAT by consultant tutors using operating sessions that are extra to the service lists is operationally and educationally viable. Furthermore, it can be implemented within a pre-determined budget. The audit of TILS has also demonstrated that the immediate clinical outcome of patients is not compromised by this type of training. PMID- 9354061 TI - The history of liver surgery. PMID- 9354062 TI - Nurse-led general surgical pre-operative assessment clinic. AB - A nurse-led Pre-operative Assessment Clinic was introduced into the General Surgical Directorate at the Royal Hallamshire Hospital in August 1993 to provide a general medical and anaesthetic pre-operative assessment, to give information, both written and verbal, to patients about their operations and to identify social problems which might delay discharge. Patients were seen either at the time of their original clinic appointment or subsequently on a recall basis, prior to admission to hospital. The clinic was run by two out-patient nurses working to agreed protocols. Patients were given written information leaflets in addition to a verbal explanation about their admission and operation. In order to assess the effectiveness of the clinic, a group of patients attending during the first year were compared with similar patients who did not undergo pre-operative assessment. A questionnaire of satisfaction with information received was completed and compliance with the clinic protocols was recorded. In the pre operative assessment group, patients were more satisfied with the amount of information received and there were significantly fewer cancellations due to unforseen medical problems in this group. Eighteen per cent of investigations were repeated unnecessarily because the reports were unavailable or considered out of date. However, only 1% of patients pre-operatively assessed had their operations cancelled after their admission, compared with 6% of non pre operatively assessed patients. Given the total number of patients admitted for surgical procedures per year, the introduction of comprehensive pre-operative assessment for all patients could result in a substantial reduction in cancellations following admission. PMID- 9354064 TI - Surgical audit: the junior doctors' viewpoint. AB - Clinical audit is now an integral part of surgical practice. The Royal College of Surgeons of England has stated that all members of the clinical team should participate in the clinical audit. Junior surgical doctors from the West Midlands Region were interviewed, using a questionnaire, to investigate their attitudes towards audit. Sixty-eight junior doctors were interviewed from six hospitals. Sixty-one (90%) felt that audit was necessary in routine surgical practice, 21 (31%) had presented data at audit meetings and only three (4%) were allocated study periods to prepare for their audit topics. In those hospitals in which all elective work was cancelled, there was a higher attendance rate at audit meetings by junior doctors. Many junior doctors felt that they lacked direction from senior colleagues and had not received any formal training in clinical audit. Clinical audit should become part of undergraduate and postgraduate medical training and junior doctors need to take a more active role in clinical audit. PMID- 9354063 TI - Improving transfusion practices in a busy teaching hospital. AB - A retrospective audit of blood transfusions was carried out in Cardiff Royal Infirmary (CRI) to investigate how efficiently blood was requested and used by the various clinical directorates. Excessive crossmatch/transfusion (C/T) ratios were found for a number of operations. In an attempt to improve practices, a pilot study was carried out between the Department of Haematology and the largest single requesting group, the Orthopaedic Directorate. As a result of a preliminary retrospective audit, crossmatch guidelines were revised, with more reliance on the group and antibody screen (G & S) for low-risk operations. A subsequent prospective audit showed major reductions in crossmatch requests and a general decrease in C/T ratios to very efficient levels without any patient morbidity. Blood was freed for urgent use elsewhere, and significant cost improvements resulted. This study, using the orthopaedic surgery department as a model, shows the value of inter-departmental audit and supports the experience of other centres using similar methods to make considerable savings in the amount of blood crossmatched unnecessarily. PMID- 9354065 TI - Surgical decompression of thoracic outlet syndrome; is it a worthwhile procedure? AB - During a 20-year period from 1974 to 1994, 37 thoracic outlet decompressions were performed. There were 28 females and six males (ratio 5:1). The median age was 37 years (range 15-64). Symptoms were predominantly neurological in 29 limbs (78%), arterial in five limbs (14%) and venous in three limbs (8%). Limb pain and paraesthesia were the most common symptoms. Surgical decompression was performed via a supraclavicular approach in 24 limbs (65%) and a transaxillary approach in 13 limbs (35%). A cervical rib was excised in 21 limbs (57%), a first rib in 10 limbs (27%), a cervical and first rib in one limb (3%) and a cervical band in five limbs (13%). Arterial reconstruction was only required in three limbs (8%). There were a total of four complications (11%). The outcome of surgical decompression was assessed by using a questionnaire completed by the patient. Overall 27 patients (87%) felt that the operation was worthwhile. These results show that surgical decompression for thoracic outlet syndrome is a worthwhile procedure and is associated with relatively few complications. PMID- 9354067 TI - The incidence of open appendicectomy, herniorrhaphy and cholecystectomy in four hospitals of the underdeveloped world and a discussion on the possible role of minimal access surgery. AB - The objective of this study was to determine the frequency of three open operations in four East and Central African hospitals. Data were collected by the author over a period of 20 years. The results show that appendicectomy and cholecystectomy are uncommon operations in the four hospitals, with an incidence of less than 1% and less than 0.35% respectively, with little change over the years. In the light of these findings, the uncertain role of minimal access surgery is discussed, and the literature reviewed. PMID- 9354066 TI - Complications following cholecystectomy. AB - Laparoscopic cholecystectomy is considered the gold standard for cholelithiasis. Nevertheless possible complications must not be underestimated. In this department, from 1 July 1991 to 30 November 1995, 1005 patients with cholelithiasis underwent videocholecystectomy. There was no peri-operative mortality. In 36 cases (3.6%) the procedure was changed to laparotomy. In four cases (0.4%) conversion was mandatory due to severe complications: in three patients while introducing a trocar (one aortic lesion, one middle colic vein injury and one visceral perforation) and in one patient due to bleeding in the hepatic hilar region. In 32 cases (3.2%) conversion was carried out electively. This was due to technical difficulties or to choledocholithiasis (22 patients), anaesthesiological problems (three cases), biliodigestive fistula (one), bile spillage from accessory hepatic ducts (three), unexpected colonic cancer (one), instrument malfunction (two cases). Twenty-four patients (2.4%) experienced post operative complications: one with pneumothorax, two with bile leakage (one bile duct damage, and one cystic duct leakage), eight with haemoperitoneum, five with subphrenic abscess, three with anaemia, three with intraparietal collections, one with bilateral basal bronchopneumonia, one with perforated duodenal stress ulcer. Of these, 11 patients (1%) underwent reintervention: five re-laparoscopies, three conversions, and three open laparotomies. This study demonstrates the safety of videolaparocholecystectomy. Complications are relatively rare and can be often dealt with conservative treatment or re-laparoscopy. Complications are often linked to insertion of a blind trocar or to the induction of a closed pneumoperitoneum. Meticulous technique or open laparoscopy minimize these risks. Conversion must not be considered a defeat but a wise decision in the face of major difficulties. Under these principles, videocholecystectomy is safe and represents the best treatment of gallbladder stones. PMID- 9354068 TI - The effect of breakfast on minor anal complaints: a matched case-control study. AB - With the objective of exploring the association between breakfast and minor anal complaints, an age, sex and pregnancy matched case-control study was carried out in the out-patient clinics at Birmingham Heartlands Hospital. Patients were selected after personal interviews using a structured questionnaire in out patient clinics. Information on age, sex, occupation and breakfast habits, as well as on haemorrhoids and anal fissure, was obtained. Patients who had haemorrhoids or anal fissure were placed in the case group; the remainder were controls. Any patient with diverticulosis, inflammatory bowel disease, colon cancer or bowel resection for any reason was excluded from the study. The main outcome measures were the odds of developing haemorrhoids or fissure in patients who did not eat breakfast. The results are based on 47 cases that were age, sex and pregnancy matched. Of the case group, 36% did not eat breakfast, compared with 11% in the control group. The analysis demonstrated a 7.5-fold increase in the odds of suffering from haemorrhoids or anal fissures in matched subjects who did not eat breakfast, with a very high level of significance (P = 0.0036). This indicates that there is a very strong association between failure to eat breakfast and haemorrhoids or anal fissure. It is anticipated that educating the public to eat breakfast would lead to a long-term fall in the incidence of anal complaints, in the attendant morbidity for the patients and in the cost to the health service. PMID- 9354069 TI - Extra-articular fractures of the proximal tibial diaphysis: their epidemiology, management and outcome. AB - In a study of 523 consecutive tibial fractures, 33 were located in the proximal extra-articular segment. There were two fracture groups with different epidemiological characteristics and prognoses: group 1 fractures were metaphyseal in location, follow low-velocity injuries and have a good prognosis with non operative treatment; group 2 injuries are high-energy diaphyseal fractures. Treatment of these latter fractures proved difficult, with all methods showing significant complications. Overall there was a 26% incidence of malunion, 7% deep infection and 7% compartment syndrome. Only 44% of patients with group 2 fractures returned to full function. Epidemiological analysis showed that proximal tibial fractures are closer to comminuted and segmental fractures in severity than to middle- and distal-third fractures. It is suggested, on the basis of our results, that they should be treated with either compression plating or closed external fixation. PMID- 9354070 TI - Three cases of prolapsed disc presenting as 'twisted ankles'. AB - We have recorded three cases of patients who initially were referred to the orthopaedic department with ankle problems. One of these was actually being managed with a full below-knee walking plaster. All of these patients proved to have prolapsed intervertebral discs with appropriate symptoms, and signs at examination and on computerized tomography (CT). Incorrect management protocols resulted in possible morbidity or at least in a delay in correct diagnosis. In summary, it is recommended to remember nerve root compression as a cause for ankle pain and for lack of abductor control of the ankle joint. This emphasizes the need for an adequate history and examination. PMID- 9354071 TI - Fine needle aspiration cytology of head and neck lesions: an early experience. AB - Fine needle aspiration cytology (FNAC) was performed on 91 patients with lesions of salivary glands and cervical lymph nodes. The results were compared with histopathology in 58 patients who subsequently underwent surgery. Of a total of 105 aspirates, a definitive cytodiagnosis was possible in 88 (83.8%). An overall sensitivity of 77.7% and specificity of 93.3% were obtained when the cytological results were correlated with histopathology in 48 cases. The figures for salivary glands were 40.0 and 88.9%, and for cervical lymph nodes were 92.3 and 100.0% respectively. The incorrect diagnosis of some aspirates did not adversely affect the subsequent management of these cases. Fine needle aspiration cytology has a role in the diagnosis and management of a variety of head and neck lesions, although it should be used, along with other investigations, not as a substitute but as an adjunct to sound clinical judgement. The limitations of the procedure, including occasional errors of cytodiagnosis, should be borne in mind. The use of this technique by otolaryngologists has been limited, and we advocate that it becomes a more integral part of the diagnostic work-up in head and neck practice. PMID- 9354072 TI - Day case adenoidectomy: is it acceptable to parents? AB - In this prospective, randomized and controlled study, 64 children were randomly allocated to in-patient or day case adenoidectomy. The pre- and post-operative concerns of the parents of both groups were measured and compared. No differences in the degree of concern was detected between the two groups. All of the parents from the day case group approved of day care, and 57% of the in-patient group would have preferred day case care. PMID- 9354073 TI - Blood loss following extraction of deciduous teeth under general anaesthetic. AB - The potential for significant blood loss following deciduous tooth extractions has been a factor in treatment planning in dental care for children. This paper reports the results of a study which aimed to quantify normal blood loss following extraction of deciduous teeth from medically fit children under general anaesthetic. Blood loss from a group of 50 children aged 3 to 5 years, having at least one molar tooth removed, was determined by comparing the amount of haemoglobin in a sample of the patient's whole blood with a known dilution of blood collected during surgery and recovery. Total blood loss ranged from 2.5 to 57 mL, with the maximum corresponding to deciduous clearance of (the removal of all deciduous teeth from) the upper arch together with the lower molars for a 3 year-old child. Clinical judgement would suggest that the maximum potential blood loss resulting from a complete dental clearance (20 teeth) from an otherwise fit child in this age group is unlikely to be greater than 100 mL. It is concluded that the risk of inducing hypovolaemic shock from this procedure is much lower than the risks of using repeated administrations of general anaesthesia to facilitate a staged dental clearance. PMID- 9354074 TI - Air gun injuries in children: a continuing cause for concern. AB - Air guns are commonly used for sporting entertainment. The inappropriate use of these weapons often leads to injury. In general, trauma inflicted by air weapons is trivial. However, the potential for more serious and fatal injuries is significant. We report two cases of serious air gun injury in children and a review of the relevant legislation covering the use of air weapons in the UK. Enhanced public education and awareness as well as coordinated policies by organizations involved in the care of children is desirable to prevent future calamities involving air weapons. PMID- 9354075 TI - Primary malignant fibrous histiocytoma of the jejunum: report of a case and review of subject. AB - We report a new case of primary malignant fibrous histiocytoma of the jejunum. Malignant fibrous histiocytoma (MFH) occurs most commonly in the extremities and trunk, but rarely in visceral organs. However, only eight cases of primary tumours involving the small intestine, including the present, have been described. This case report documents the appearance of malignant fibrous histiocytoma as a primary lesion of the intestinal wall in a patient with a 2 month history of dyspepsia, weight loss and unspecific abdominal pain. The final diagnosis was based on the pathological report of the surgical specimen. Emphasis is placed on the clinical signs, radiological studies and pathological findings. The literature on MFH of the jejunum is also reviewed. Malignant fibrous histiocytoma is considered an aggressive tumour, and the treatment of choice is complete surgical excision. Adjuvant chemotherapy or radiation is recommended mainly in those patients in whom there is vascular or lymphatic infiltration. PMID- 9354076 TI - Distal splenocaval shunt as an alternative to distal splenorenal shunt in a patient with a retro-aortic left renal vein. AB - A 19-year-old patient with schistosomiasis and portal hypertension presented with bleeding oesophageal varices. A selective distal splenorenal shunt was planned. At surgery a retro-aortic left renal vein prevented a classical splenorenal anastomosis, and a distal splenocaval anastomosis was performed which is clinically patent at 18 months. PMID- 9354077 TI - Duodenal obstruction from chronic pancreatitis. AB - The fibrosclerosing process of the pancreas in the chronic pancreatitis may constrict not only the pancreatic duct but also the bile duct, splenoportal venous system and duodenum. In our retrospective study we analysed 24 patients with duodenal obstruction associated with chronic pancreatitis. Duodenal obstruction was suspected whenever repeated vomiting occurred or large volumes of nasogastric aspirate were obtained. The diagnosis was confirmed by barium meal and endoscopic examination. Duodenal obstruction was relieved by gastrojejunostomy in eight patients, gastrojejunostomy and vagotomy in eight patients, gastroduodenostomy and vagotomy in two patients, vagotomy with Finney pyloroplasty in one patient, duodenoplasty with vagotomy in one patient and Whipple procedure in four patients. We concluded that vagotomy and gastroenterostomy are the procedures of choice. Bypass surgery is helpful to relieve the obstruction of the common bile duct and pancreatic duct. Whipple procedure should be reserved for the small duct form of chronic pancreatitis and for the cases in which there is high suspicion of malignancy. PMID- 9354078 TI - Posterior wall perforation of duodenal ulcers. Report of two cases and survey of the literature. AB - Retroperitoneal perforation of duodenal ulcer is a rare condition. Diagnostic difficulties lead to high mortality. Two cases are presented in this report. PMID- 9354079 TI - Reactive oxygen species and antioxidants in signal transduction and gene expression. AB - Reactive oxygen species (ROS) are produced by cellular metabolic reactions, and have been implicated in the pathogenesis of several diseases, including atherosclerosis, cancer, and Alzheimer's disease. Interestingly, clinical and epidemiologic studies have, in some cases, indicated that antioxidant nutrients may be effective in disease prevention. However, the efficacy of specific antioxidants in disease prevention is often both controversial and inconclusive. In an effort to elucidate the role of ROS and antioxidants in disease development and prevention, the chemistries of ROS and antioxidants have been examined extensively. Recently, molecular and cellular approaches have demonstrated that ROS and antioxidants can directly affect the cellular signaling apparatus and, consequently, the control of gene expression. This new research provides the link between ROS and antioxidant chemistries and the mechanisms of disease processes and prevention. This review illustrates how ROS function as potential intracellular and extracellular signaling molecules and how antioxidants can affect this process. PMID- 9354080 TI - Nutritional implications of xerostomia and rampant caries caused by serotonin reuptake inhibitors: a case study. AB - Serotonin reuptake inhibitors, such as fluoxetine, fenfluramine, and dexfenfluramine, are frequently used to treat obesity, depression, and bulimia. A common side effect of these medications is xerostomia, or dry mouth. A case study demonstrating the impact of drug-induced xerostomia on oral health and subsequent nutrition implications is presented. Rampant caries can result from a combination of xerostomia and inappropriate dietary and oral hygiene habits. Preventive dietary and dental guidelines are presented to assist nutrition and dental professionals in treating and counseling patients with xerostomia. PMID- 9354081 TI - High-dose vitamin supplements for cigarette smokers: caution is indicated. AB - Results from human intervention studies indicate that smokers should avoid using high-dose dietary beta-carotene supplements. In addition, problems may exist for smokers with high-dose vitamin E supplements. PMID- 9354082 TI - Effect of calcium supplementation is greater in prepubertal girls with low calcium intake. AB - In a recent calcium supplementation trial, prepubertal girls with spontaneous calcium intake below 900 mg/day, when given additional calcium, had greater increases in bone mineral density than other girls. This lends support to the recently recommended Adequate Intake (AI) of 1300 mg/day calcium for American and Canadian children, although more studies on long-term effects of this intake level are still needed. PMID- 9354083 TI - Confirmation: chromium levels in serum, hair, and sweat decline with age. PMID- 9354084 TI - gamma-Tocopherol: an efficient protector of lipids against nitric oxide-initiated peroxidative damage. AB - Nitric oxide released by macrophages during inflammation reacts with active oxygen to form peroxynitrite. Peroxynitrite nitrates protein and peroxidizes lipids. gamma-Tocopherol traps peroxynitrite and is more effective than alpha tocopherol in protecting lipids against such peroxidation. PMID- 9354085 TI - Pleasantness, activation, and sex differences in advertising. AB - Advertisements in men's, women's, girls', and boys' magazines (n = 38,195 words) were scored objectively in terms of 15 measures of linguistic style, e.g., use of common words, use of long words, use of specific words and emotional tone (pleasantness and activation, as measured by the Dictionary of Affect). There were several sex- and age-related differences among advertisements from different sources. Advertisements from boys' magazines were extremely active, those from women's and girls' magazines were shorter and unusually pleasant. In two follow up studies (N = 122 volunteers), objective emotional measures of advertising text proved to be related to ratings of persuasion and of success of appeal for individual advertisements. The most preferred advertisement for women was pleasant and active, that for men unpleasant and active. When men and women created advertisements, women's were shorter and more pleasant. PMID- 9354086 TI - Type A behavior and the five-factor model of personality. AB - The hypothesis that people classified as Type A and Type B have different personality profiles based on five major personality factors (Neuroticism, Extraversion, Openness, Agreeableness, and Conscientiousness) was tested using the Student Jenkins Activity Survey and the Revised NEO Personality Inventory. Results based on discriminant function analysis of data from 243 psychology undergraduates (105 males and 138 females) strongly supported the hypothesis indicating that Type A and Type B groups have significantly different Revised NEO Personality Inventory profiles and that the standardized discriminant function coefficients were large for Agreeableness and Conscientiousness and moderately large for Extraversion. PMID- 9354088 TI - Relations of early maternal employment and attachment in introvertive and extravertive adults. AB - The present study examined attachment scores of adult children whose mothers were employed and how maternal employment varied as a function of children's personality styles. Children's extraversion was expected to moderate the effects of maternal employment on their attachment as adults. Responses of 106 undergraduates were obtained on 3 measures, the Eysenck Personality Inventory, the Adult Attachment Scale of Collins and Read, and the Adolescent Relationship Scales Questionnaire of Scharfe and Bartholomew. A median split was performed to divide subjects into those scoring High and Low on Extraversion. Subjects were then grouped on the basis of their mothers' employment status during the subjects' infancy (Full-time, Part-time, Non-employed). Subjects high on Extraversion seemed to show more adverse attachment consequences in adulthood following full-time maternal employment during infancy. Adults who scored high on extraversion may have been more comfortable with continual maternal presence during infancy, while those more introverted as adults may have adapted better to the periods of separation associated with infant day care. PMID- 9354087 TI - A Canadian-Dutch comparison of teachers' burnout. AB - Scores on burnout among 631 Canadian and 1,180 Dutch teachers were compared with various demographic variables (sex and age) and factors related to work (experience in teaching, type of school, and number of hours employed). Burnout was assessed with the Maslach Burnout Inventory of three dimensions, Emotional Exhaustion, Depersonalization, and Personal Accomplishment. Analysis indicated that, over-all, Canadian teachers reported higher scores on Emotional Exhaustion and Depersonalization than their Dutch peers. Differences in the number of hours employed were also significant: full-time Canadian teachers scored higher on Depersonalization than their Dutch colleagues. Across countries, sex and type of school appeared significantly related to burnout. Male teachers rated higher on Emotional Exhaustion and Depersonalization than the women. Especially with regard to the attitudinal components of burnout, i.e., Depersonalization and Personal Accomplishment, secondary school teachers reported higher scores than elementary school teachers. Age was not significantly related to measures. PMID- 9354089 TI - Use in 1992 of amphetamines among Miami's public school students. AB - This analysis examined self-reported amphetamine use by 473 adolescents in Dade County, Florida public schools in 1992. Significant factors which increase the probability of use include peers' use of amphetamines and the fact that the adolescent is a female. PMID- 9354090 TI - Occupational stress among secretarial personnel at the University of Transkei. AB - The objective was to explore how much 22 secretarial personnel (M age = 36.0 yr., SD = 4.8 yr.) at the University of Transkei, 14 of whom were unmarried, experienced stress in their work. Scores on a 20-item questionnaire indicated no occupational stress. PMID- 9354091 TI - Reconsideration of the Locke-Wallace Marital Adjustment Test: is it still relevant for the 1990s? AB - The Locke-Wallace Marital Adjustment Test has been in use for over thirty years despite the development of other scales. The role of the test is discussed in terms of theoretical and practical concerns in the measurement of marital satisfaction. Major criticisms are briefly reviewed and empirical questions are identified. These questions are addressed using archival data on four samples totalling 281 couples. The test possesses adequate reliability and good criterion related validity. A single factor was identified for both men and women. Ten items discriminated in all analyses. Minor changes in the scoring procedure were suggested for two items in response to some criticisms. These changes did not affect the psychometric properties. The alternative scoring system proposed by Hunt was also evaluated and a cut-score was identified. Continued use of the test is justified in general contexts where the broadly based definition of adjustment is appropriate. More comprehensive measures of adjustment and satisfaction and simpler measures of marital quality still leave a role for this 15-item rapid assessment measure of marital adjustment. PMID- 9354092 TI - Coping and mood in HIV-positive women. AB - To date, little empirical research on the association of coping style with mood in HIV-positive women has been carried out. The extant literature on HIV-positive men suggests that active coping is related to diminished distress while avoidant coping is associated with elevated distress. Previous research with HIV-positive women has not consistently confirmed these relationships. To add to this literature, scores from a sample of 145 HIV-positive women who completed the Ways of Coping Questionnaire and the Mental Health Inventory were analyzed. Correlations indicated that an escape-avoidance strategy was associated with more negative emotions. Other strategies related to negative emotions included accepting responsibility and a self-controlling approach. These findings are consistent with those previously reported for HIV-positive men, suggesting that similar kinds of coping strategies may be associated with positive and negative moods among HIV-positive men and women. PMID- 9354094 TI - Reliability of social and personal competence measures for adolescents. PMID- 9354093 TI - Relationships between verbal and nonverbal fluency measures: implications for assessment of executive functioning. AB - This experiment evaluated the relationship between verbal and nonverbal fluency measures commonly employed in neuropsychological assessment. Three fluency measures, the Controlled Oral Word Association Test, the Design Fluency Test, and the Ruff Figural Fluency Test, were administered to a sample of 61 men and 73 women in college. Analyses indicated that scores on the Controlled Oral Word Association Test were significantly correlated with scores on the Fixed but not the Free Condition of the Design Fluency Test. Scores on the Ruff test were significantly more closely correlated with the Fixed versus the Free Condition, although they were also correlated with the total scores on the Design Fluency Test. Differential correlations are discussed in terms of the varying structure inherent in the measures of fluency and their presumed taxing of frontally mediated executive processes. PMID- 9354095 TI - Dating, social avoidance and distress. AB - For 200 undergraduates association of dating and social avoidance and distress were explored. Conditioned anxiety, physical attractiveness, skills in dating, proximity, and apprehension about dating differentiated between high and low scorers on social avoidance and distress. PMID- 9354096 TI - Job satisfaction and suppression of emotions. PMID- 9354097 TI - Sex differences on the MMPI-1 and MMPI-2. AB - This study compared sex differences on the MMPI-2 to those previously found on the MMPI-1. Statistically significant differences were found with men (n = 66) showing higher mean raw scale scores on antisocial (men = 10.00, women = 7.15), authority problems (men = 4.58, women = 3.14), admission of addiction (men = 3.58, women = 2.74), amorality (men = 2.30, women = 1.61), and Type A (men = 8.24, women = 7.71). Women (n = 132) showed higher mean raw scale scores on Hypochondriasis. Depression, Hysteria, Paranoia. Depression subscale, N affection, Health Concerns, and Somatic Complaints. With a few exceptions, these general patterns are consistent with previous research. PMID- 9354098 TI - Personality and criminal profile of adolescent sexual offenders, general offenders in comparison to nonoffenders. AB - 16 adolescent male sex offenders and 13 general offenders were compared with 13 nonoffenders on psychometric tests to investigate differences in their general intelligence, personality, and criminal attitudes. There were no significant differences in general intelligence amongst the groups. Examination of personality scores and criminal attitudes showed that the sex offenders were more socially isolated, more assaultive, and more resentful than the general offenders. Present results, if replicated with larger samples, suggest treatment of adolescent sex offenders in a manner similar to that used in the treatment of adult sex offenders. PMID- 9354099 TI - Use of age-adjusted suicide rates in time series studies in Canada. PMID- 9354100 TI - Unprotected anal intercourse in gay men: the resolution to withdraw before ejaculating. AB - 734 gay men who had recently engaged in unprotected anal intercourse reported self-justifications they used at the time. A common self-justification involved a resolution to withdraw before ejaculating. Compared with other self justifications, this resolution was associated with a "last minute" rather than a "premeditated" decision to have unprotected anal intercourse, suggesting that the resolution derived just from "heat of the moment" thinking. Implications for AIDS education are discussed. PMID- 9354101 TI - Unemployment and psychological health among Hong Kong Chinese women. AB - The impact of unemployment on psychological health, indexed by the General Health Questionnaire, was studied in 86 unemployed and 79 employed Chinese women in Hong Kong. As with findings reported in the West, the present results showed that the unemployed participants were more disturbed than their steadily employed peers. In addition, the prevalence rate of disturbance (54%) observed in the present sample of unemployed women is comparatively higher than those of Western samples reported previously. Implications of these preliminary findings for research on psychological aspects of unemployment among Hong Kong Chinese were discussed. PMID- 9354102 TI - The sex ratio in American Indian suicides. PMID- 9354103 TI - Jewishness and well-being: specific identification and general psychological adjustment. AB - The study examined the relation of nine factors describing the expression of Jewish identification with three scales related to aspects of psychological well being (Belonging, Optimism and Self-acceptance) among 177 American Jews, ages 22 to 40 years. Regression analyses indicated a small but significant, positive relation between scores on each of the well-being scales and two aspects of Jewish identification, activism in mainstream Jewish organizations (Beta weights .15 to .23) and religiosity (.19 to .21). These expression of Jewishness accounted for 7 to 11% of the variance in reported well-being. PMID- 9354104 TI - Self-reinforcement, gender-role, and sex of participant in prediction of life satisfaction. AB - There have been conflicting results regarding sex differences and gender role in predicting life satisfaction and no research assessing the relationship between life satisfaction and self-reinforcement. These relationships were evaluated by administering to 182 undergraduates the Bem Sex Role Inventory, the Satisfaction with Life Scale, and the Frequency of Self-reinforcement Questionnaire. A regression analysis showed significant effects for self-reinforcement and gender role in the prediction of life satisfaction. No main effect was found for sex of participant and there were no significant interactions. Scores on measures of self-reinforcement and life satisfaction were moderately correlated. PMID- 9354105 TI - Burnout syndrome and type A behavior in nurses and teachers in Sicily. AB - Burnout and Type A behavior were studied in two groups of 50 teachers (26 working in high school and 24 in junior high school) and 50 nurses living and working in Sicily. Each group was composed of 19 men and 31 women. A revised version of the Maslach Burnout Inventory and the Adult and Adolescent Type A Behavior Scale Revised Form 1 were used to measure burnout and Type A behavior, and a scale of job satisfaction was given. Analysis shows higher scores on stress for nurses, related to the low social acknowledgment of their job. Among nurses, Type A scores were correlated positively with scores on burnout and negatively with ratings of job satisfaction. The teachers showed greater compatibility with their work which may reflect that in Sicily, the teaching profession is still held in high regard. PMID- 9354106 TI - Psychopathy and substance use in adolescent male offenders. AB - Previous research with incarcerated adult male offenders yielded correlations of .28 to .40 between measures of substance abuse and psychopathic characteristics. This comorbidity was replicated in 40 incarcerated adolescent male offenders. Drug and alcohol use was significantly correlated with the behavioral characteristics of psychopathy (r = .48 and .41, respectively). PMID- 9354107 TI - Measuring quality of nursing home service: residents' perspective. AB - An inventory was developed to measure residents' perceptions of the quality of nursing home service. The content domain and dimensions of the inventory were derived from actual comments of nursing home residents. Independent studies employing a multiple-facility sample of 103 residents and 194 residents from a single institution supported a four-factor structure of the quality of nursing home service--Staff and Environmental Responsiveness, Dependability and Trust, Food-related Services and Resources, and Personal Control. The data provide preliminary support for the measure's reliability and validity so it may be used to study the antecedents and consequences of quality in nursing home service from the residents' perspective. PMID- 9354108 TI - Perceived control by "powerful others" in paranormal healers. AB - 49 paranormal healers working by laying-on of hands (direct healing) and healing at a distance were compared with 56 nurses and a control group of 73 randomly selected persons on their locus of control and especially on the subscale Powerful Others of Levenson. The paranormal healers scored external in locus of control, and their scores differed significantly from those of the other two groups tested on the Powerful Others subscale. This finding may be associated with the dependent position of paranormal healers since their profession is tolerated by medical doctors and the law. PMID- 9354109 TI - Self-narcissism and interpersonal attraction to narcissistic others. AB - In the first of a two-part study, 172 participants completed a questionnaire on personality and career preferences. Items from the Narcissism Scale of the Million Clinical Multiaxial Inventory were embedded in this questionnaire as well as a series of bogus items. In the second session conducted three weeks later, participants watched a videotaped dramatization of either a male or female enacting a narcissistic role and completed a modified version of the First Impressions Questionnaire and an item assessing mood. The narcissism scores of participants obtained during Part 1 were paired with their respective ratings of the target person on the modified-First Impressions Questionnaire and mood. Contrary to predictions participants' scores on narcissism did not affect their first impressions of persons enacting a narcissistic role. Participants who viewed the male role player rated him as less attractive than those who watched the female, while participants who watched the female target reported greater negative mood scores than those who watched the male target. PMID- 9354110 TI - Increasing return rates to a mail survey among health educators. AB - This study indicated a combination of methods increased the return rate (80%) in a national survey of health educators (n = 546). Also, use of a $1.00 bill incentive was not significantly more effective than no incentive, and the rate of double responses in this anonymous mail survey was very low (1%). PMID- 9354111 TI - Yoga breathing through a particular nostril increases spatial memory scores without lateralized effects. AB - Uninostril breathing facilitates the performance on spatial and verbal cognitive tasks, said to be right and left brain functions, respectively. Since hemispheric memory functions are also known to be lateralized, the present study assessed the effects of uninostril breathing on the performance in verbal and spatial memory tests. School children (N = 108 whose ages ranged from 10 to 17 years) were randomly assigned to four groups. Each group practiced a specific yoga breathing technique: (i) right nostril breathing, (ii) left nostril breathing, (iii) alternate nostril breathing, or (iv) breath awareness without manipulation of nostrils. These techniques were practiced for 10 days. Verbal and spatial memory was assessed initially and after 10 days. An age-matched control group of 27 were similarly assessed. All 4 trained groups showed a significant increase in spatial test scores at retest, but the control group showed no change. Average increase in spatial memory scores for the trained groups was 84%. It appears yoga breathing increases spatial rather than verbal scores, without a lateralized effect. PMID- 9354112 TI - Predictor variables of clergy pedophiles. AB - File data on familial traits, past sexual experience as a victim, and other traits identified in the literature as leading toward pedophilia, were summarized for 10 convicted clergy pedophiles to construct a set of variables possibly useful for screening. Further research is underway to identify trauma in early life and those personality-related variables current studies indicate as relevant. PMID- 9354113 TI - Loneliness: a predictor of health perceptions among older Korean immigrants. AB - This study examined whether loneliness predicts health perceptions, i.e., emotional and physical health, in a sample of 174 older Korean immigrants living in a metropolitan area. Scores on the revised UCLA Loneliness Scale predicted scores on the Life Satisfaction Index-Z and perceived physical health in the older Koreans, but not scores on the Symptom Pattern Scale. PMID- 9354114 TI - Health and socioeconomic indicators in psychiatric catchment areas with divergent suicide rates. AB - Differences found in the incidence of suicide between the psychiatric catchment areas of the Karolinska Hospital in Stockholm were investigated in relation to health and socioeconomic indicators during the study period 1990-1994. The hypothesis of the study was that negative socioeconomic indicators and psychosocial and health indicators denoting less favourable socioeconomic status may negatively influence the suicide rate of the demographic units in this cross sectional study. The incidence of suicide between the areas was significantly different and increasingly divergent in the last year of the study period. The area with a higher proportion of suicides had also an increased proportion of individuals who retired early, lower life expectancy at birth, higher non employment, lesser income among the employed, less public expenditure for education, less proportion of home ownership, and a higher proportion of persons bound to one-room dwellings. Disregarding the influence of ethnicity (there were no statistically significant differences on immigrants' suicide between the areas) as well as in the availability of psychiatric care (assuming that similar quality of psychiatric care was provided by both sectors), or other demographic indices commonly shared by the areas, the possibility of strong effects of unfavourable health and socioeconomic indices appeared relevant for the explantation of an increased incidence of suicide. The findings provide new empirical contradiction to the socioeconomic hypothesis of the incidence of suicide, which postulated that populations with higher socioeconomic status may have increased suicide rates. PMID- 9354115 TI - Intervention-based assessment: rates of evaluation and eligibility for specific learning disability classification. AB - Intervention-based assessment, a systematic form of prereferral intervention, represents a viable alternative to "test and place" models for identifying and teaching children with a variety of learning-related problems in schools. Data obtained from 13 schools participating for a third year in a pilot study of statewide implementation of intervention-based assessment suggested that, in comparison to a prior prereferral intervention model, fewer children are evaluated and found eligible for special education. Of those children receiving intervention-based assessment, a slight decrease occurs in the percentage classified as specifically learning disabled. PMID- 9354116 TI - Anger management training in prison inmates. AB - Inflicting harm on others after a perceived wrong is called revenge and has been implicated in a wide range of criminal and antisocial behaviors. Revenge is defined as a retaliatory act and may be ruled out when antecedent to instrumental aggression if hurting someone is secondary to the primary goal of acquisition. Revenge is considered the impetus for reactive aggression, however, if the primary goal is to hurt someone. 26 male inmates were chosen for training in anger management using cognitive behavioral methods. Selection of inmates was based on their history of reactive aggression. As predicted, inmates showed a significant reduction in posttest scores on the Vengeance scale. PMID- 9354117 TI - Alexithymic characteristics of bulimia nervosa in diabetes mellitus with end stage renal disease. AB - This study examined the clinical characteristics including stress-related factors of eating disorders in a sample of 312 diabetic patients with end-stage renal failure. The prevalence rate of bulimia nervosa was 5.1% (16 of 312 patients). The 16 patients with bulimia nervosa were 8 men and 8 women over 58 years old. Looking at the subjects by cause of end-stage renal failure, those with diabetes mellitus exhibited significantly higher prevalence rate of bulimia nervosa than two nondiabetic groups (diabetes 10%; nephritis 1.6%; others 1.9%). As for the association of bulimia nervosa and stress-related factors, end-stage renal failure patients with diabetes who exhibited bulimia nervosa showed significantly higher scores on a measure of alexithymia. These results suggest that, when liaison psychiatrists see diabetic patients with end-stage renal failure who exhibit bulimia nervosa, they should pay close attention to stress-related symptoms including alexithymia. PMID- 9354118 TI - Lack of medical support in HIV infection and unstable mood states. AB - This study examined what kinds of social support are related to mood states in a sample of 50 HIV-positive patients without AIDS (46 men and 4 women; M age 36.5 yr., SD = 9.8). In the early stage of HIV infection, HIV patients without AIDS may be prone to depressive symptoms although none of these HIV-positive patients' symptoms fulfilled the DSM-III-R Mood Disorders including Major Depression. The depressive symptoms were not significantly related to lack of ordinary social support such as friends and family but were significantly associated with dissatisfaction with HIV/AIDS-related medical support. PMID- 9354119 TI - A modified work locus of control scale: preliminary investigation of reliability and validity in a sample of pharmacists. AB - Items in Spector's Work Locus of Control-Scale were modified to reflect how much control the respondent feels he might have on a job as opposed to what he feels people in general have. This modified Work Locus of Control Scale was tested for reliability and construct validity. As part of a larger study, data were collected via mail questionnaires from a sample of 284 pharmacists licensed in Indiana. The Cronbach coefficient alpha for the modified scale was .88, indicating good internal consistency. The modified scale also showed some evidence of convergent validity when correlated with scores on measures of Extraversion, Conscientiousness, Neuroticism, and Job Dissatisfaction. PMID- 9354120 TI - Perceived sources of loneliness of incarcerated men. AB - The present study examined the influence of incarceration on sources of loneliness. It was hypothesized that incarcerated offenders would perceive the causes of their loneliness differently than a general population sample. 145 male offenders and 112 men from the general population who were recruited on a voluntary basis, reported the sources of their loneliness on a 15-item (yes/no) questionnaire. Analysis indicated a significant difference in the perceived sources of loneliness amongst the two populations. Also duration of loneliness, i.e., chronic vs episodic, was associated with perception of its causes. PMID- 9354121 TI - Comparison of self-assessment of premenstrual symptoms with scores on the modified Menstrual Distress Questionnaire. AB - Premenstrual symptoms were assessed in a sample of 267 women (M age 31.4 yr.) using a single self-identification question and the modified Menstrual Distress Questionnaire of Clare and Wiggins. The-self-identification question asked to what degree the women experienced premenstrual symptoms. The responses were 34 (13%) for none, 116 (43%) for slight, 99 (37%) for moderate, and 18 (7%) for severe. The mean score on the questionnaire was 23.5 (SD = 17.5). Correlations indicated significant relationships between self-identification and questionnaire scores (rho = .76, p < .001). When the women were classified according to Clare's (1983) criterion, almost a third of them assessed their symptoms differently, i.e., while they classified themselves as "nonsufferers" on the self identification question, their responses on the questionnaire identified them as "sufferers." PMID- 9354122 TI - Architects: gender-role and hiring decisions. AB - To examine architects' judgments of male and female applicants represented by the information in resumes, 204 architects, 156 men and 48 women, licensed in the state of Connecticut participated in a 2(job level) by 2(sex) between-subjects study. Architects were asked how they would rate applicants' potential (including the decision to hire) and gender-role characteristics judged on the basis of one page resumes. Architects randomly assigned resumes for one of four evaluation conditions (intern or senior architect; male or female), rated the applicant on seven job-related characteristics, e.g., technical skill, potential for advancement, and completed the Bem Sex-role Inventory as they thought items applied to the applicant. Analysis indicated that male architect respondents were more likely to hire male applicants than female applicants as senior architects and that female applicants were judged to be as masculine-typed as were male applicants. PMID- 9354123 TI - An exploration of attitudes on sexuality at a northeastern urban university. AB - This study was conducted to assess the prevalence of scores on homophobia among 104 college students at a northeastern urban university. Participants reported their attitudes regarding homosexuals and homosexual behavior on Hudson and Rickett's Index of Homophobia. The 33 men indicated more negative attitudes about homosexuals and homosexual situations than the 71 women. PMID- 9354125 TI - Need for cognition and irrational beliefs. AB - A total of 149 college students, 48 men and 101 women, completed the Irrational Beliefs and the Need for Cognition scales to assess irrationality and the tendency to engage in effortful cognitive activity. Although substantial overlap between the constructs was found (r = -.29, p < .001), the relationship was not linear. Need for Cognition appears to be a process variable, while irrational belief appears to be a content variable. Results suggest that the Need for Cognition must achieve a minimum threshold before spontaneous critical self examination can occur. PMID- 9354124 TI - Social drinking and laughter. AB - Pairs of observers monitored drinking and laughter in groups of social drinkers (N = 56). A significant correlation of .27 was found between units of alcohol consumed and laughter scores. PMID- 9354126 TI - Eighty-five years of military maxillofacial surgery in Yugoslavia and twenty-five years of the Clinic for Maxillofacial Surgery of the Military Medical Academy. PMID- 9354127 TI - Maxillofacial war injuries during the war in former Yugoslavia. AB - War wounds of the maxillofacial region occur in 5-17% of casualties. In the Clinic for Maxillofacial Surgery of the Military Medical Academy, during the war in former Yugoslavia, 482 casualties with dominant wounds in the maxillofacial region were treated. The outcome of the treatment depended on the quality and speed of the first aid and general medical aid, as well as on qualified specialist treatment. Specificity of the management of the war wounds of maxillofacial region includes: minimal debridement of soft tissues, removal of only deperiosted fragments of bony tissues, valid reconstruction of bony and soft tissues defects, correct immobilization, antitetanus prophylaxis, adequate antibiotic therapy, as well as permanent and very meticulous postoperative care. PMID- 9354128 TI - The treatment of traumatic facial bones and jaw fractures in the period 1972 1996. AB - During the last twenty-five years, in the period from January 1, 1972 to December 31, 1996, 1006 injured with facial and jaw fractures have been treated at the Clinic for Maxillofacial Surgery of the Military Medical Academy--91.05% of the injured were males and 67.98% belonged to the age group between 11 and 30. The most frequent etiologic factors were traffic accidents (46.42%) and fights (38.07%). Isolated fractures of mandible were found in 51.09% of the injured. In 70.57% of the treated the surgery was used for adequate reduction, osteosynthesis and stabilization of bone fragments, and 29.34% of the injured were treated conservatively, which involved the manual reduction and intermaxillar immobilization. The post-operative complications were observed in 51 (5%) of the injured. PMID- 9354129 TI - Monocortical osteotransplant of the iliac bone in reconstruction of mandibular gunshot defects. AB - In the Clinic for Maxillofacial Surgery of Military Medical Academy, 134 injured with mandibular defects were treated, as well as a significantly larger number of injured from the territory of former Yugoslavia, with dominant injuries of the head and neck regions. Until June 1994, smaller defects were treated using bicortical osteotransplant of the iliac bone. Due to its only partial successfulness in reconstruction of mandibular defects, monocortical osteotransplant of the iliac bone was selected for reconstruction of the mandibular defects and the results were much better compared to the other free osteotransplants. PMID- 9354130 TI - Facial nerve injuries and its treatment. AB - Twenty four patients with traumatic interruption of the conductivity of the facial nerve were treated in the period 1991-1996 at the Clinic for Maxillofacial Surgery. After topographic orientation about the site and level of the injury, the reconstruction of the nerve using nerve grafts was performed in all patients. The results were satisfactory. PMID- 9354132 TI - Prosthetic reconstruction of facial defects. AB - A seven-year experience in prosthetic reconstruction of facial defects is presented. Surgical-prosthetic treatment was performed in 25 patients. Facial defect associated with organ loss of the midface was found in 4 (16%) patients. Isolated facial or midfacial organ defects were found in 21 (84%) patients, and dominant eye loss in 11 (44%) patients. All patients were managed by facial prostheses made of rigid acrylate and soft silicon. To obtain retention and stability of facial prostheses, eye glasses were most frequently used in 32% patients, and modern methods of retention with implants in 26%. The use of implants improved prosthetic reconstruction in terms of stability and retention. PMID- 9354131 TI - Vascularized fibular graft in the reconstruction of posttraumatic mandibular defects. AB - The results of the use of the vascularized fibular graft for the reconstruction of posttraumatic defects of the mandible are presented. Different surgical procedures were applied in reconstruction of the mandibular defects in 120 wounded. In the last four years vascularized fibular graft was used in 11 mandibular reconstructions. Taking into account the survival rate of 100%, long vascular pedicle, strength, length and the possibilities of adaptation of the bone make this graft suitable for mandibular reconstruction. The functional and esthetic results obtained by the use of this graft were satisfactory, with minimal donor site morbidity. PMID- 9354133 TI - Implants in management of gunshot injuries of teeth, facial bones and jaws. AB - The use of implants in management of gunshot injuries of teeth, face and jaws presents novelty that is manifested in primary surgical treatment, reduction and fixation of bone fragments and teeth, replacement of lost parts of bone tissues as well as preparation for definitive prosthetic management. At the Clinic for Maxillofacial Surgery of the MMA 173 implants of different types and purposes have been placed in the period 1991-1997, in rehabilitation of the patients injured during the civil war. Although this number of placed implants is significant, the conclusive estimate about benefit of using implants may be obtained only after a longer period of time. Early results are satisfactory. PMID- 9354134 TI - Two-component transdental implants. AB - Transdental implantation is a procedure of tooth stabilization in maxillary and/or mandibular bone. Indications for use of these implants are root fractures, chronic periapical inflammations and periodontal disease. Transdental implants have been used in the Clinic for Maxillofacial Surgery since 1977. In the period of 20 years of treatment 490 two-component transdental implants have been inserted and failure rate was 9% (43). Considering our experience and that of other authors we can conclude that with proper indications and sophisticated surgical technique the use of this type of implants has favorable survival rate. PMID- 9354135 TI - How we have treated parotid gland tumors. AB - Since the Clinic was established in 1972 to the end of 1994 at the Clinic for Maxillofacial Surgery of the Military Medical Academy 782 patients with tumors of the parotid salivary glands of glandular origin were treated. This number included only patients with sufficient data for the relevant analysis. There were 659 (85.53%) benign and 123 (14.47%) malignant tumors. They are rare in the age 0 20. Their incidence increases after the third decade, reaching the maximum in the sixth decade. The most frequent tumors were of 3 cm diameter (36.24%). The average size of tumors varied from 3 to 6 cm in diameter (29.36% of cases). Tumors larger than 15 cm are extremely rare. All patients were surgically treated: subtotal parotidectomy was done in 55.23%, total in 38.61% and radical in 6.16%. Transient facial nerve injuries were recorded in 18.63% of cases (mainly at total parotidectomy). Out of 94 patients with malignant tumors of the parotid gland, 78.13% survived for 2 years and 48.89% for 5 years. Recurrence rate for benign parotid tumors was 2.38%, and for malignant 17.62% (after total parotidectomy) and 26.12% (after the radical one). PMID- 9354136 TI - Bimaxillary osteotomies in correction of dentofacial deformities. AB - In 342 patients with dentofacial deformities cephalometric analyses were preoperatively performed. A study of occlusion on cast models as the simulated new position of the facial soft tissues was done on the photographs. Surgical treatment using bimaxillary osteotomies for correction of dentofacial deformities was done in 38 patients: 31 with prognathism and maxillary hypoplasia, 5 with facial asymmetry and laterognathism and in 2 with otomandibular dysostosis. The LeFort I type osteotomy was used for the upper jaw and Obwegeser technique for the lower jaw. Intermaxillary and circumzygomatic immobilization were removed 8 10 weeks after the operation. After that, the patients retained the individual chin traction from 2 to 3 months. The complete relapse occurred in one patient and prolonged loss of the mandibular nerve sensibility in one. PMID- 9354138 TI - Craniofacial surgery at the Clinic for Maxillofacial Surgery of the Military Medical Academy. PMID- 9354139 TI - The use of implants in dentistry. PMID- 9354137 TI - Current concepts in oral implantology. PMID- 9354140 TI - Reconstruction of a face destroyed by an explosion. AB - We have followed up the course of the facial reconstruction in a 12 year old boy with totally destroyed middle and lower third of the face when the fuse for initial mine activation exploded in his mouth. The boy has undergone more than 20 operations within the 7-year period and his rehabilitation has not been completed, yet. PMID- 9354141 TI - BCT Implant System. PMID- 9354142 TI - Emergency equipment for maxillofacial surgery. PMID- 9354143 TI - Development and analysis of oxygen-sensing probes for in situ monitoring of unsaturated solid-state biodegradation processes. AB - Oxygen (O2) sensors have been developed for in situ measurement of O2 in high solids degradation processes. The O2 sensor has been shown to withstand the corrosive environment of the biodegradation process with high humidity and temperatures exceeding 60 degrees C. Calibration tests prior to and after in situ operation showed the sensor to perform accurately and reproducibly after 71 days of in situ operation. A linear relationship between O2-sensor output and water vapor pressure was confirmed through calibration experiments. To compensate for the effect of water vapor pressure on O2-sensor measurements, O2 concentration was expressed on a dry air basis using the confirmed linear relationship. In situ O2-sensor output expressed on a dry air basis was found to follow trends of gas samples analyzed by gas chromatography with good agreement. PMID- 9354144 TI - The application of exposure-based criteria in developing alternative primary ambient ozone standards. AB - The form of the primary National Ambient Air Quality Standard (NAAQS) for ozone has been a matter of some concern in recent years. The Environmental Protection Agency is reviewing the ozone NAAQS and has proposed a set of alternative standards that address some deficiencies. However, the proposed alternatives are all based upon the ozone concentrations occurring at the peak monitor, which may not be representative of the air quality and population exposure throughout a nonattainment area. Representativeness could be improved by converting the form of the standard from an exceedance-based metric (with x allowable exceedances per year, on average) to one based upon exposure. Three approaches to defining an exposure-based standard are developed here. Data from the San Francisco Bay area are used to illustrate the proposed standards. PMID- 9354145 TI - Characterization of air quality problems in five Finnish indoor ice arenas. AB - The air quality in five Finnish ice arenas with different volumes, ventilation systems, and resurfacer power sources (propane, gasoline, electric) was monitored during a usual training evening and a standardized, simulated ice hockey game. The measurements included continuous recording of carbon monoxide (CO), nitric oxide (NO), and nitrogen dioxide (NO2) concentrations, and sampling and analysis of volatile organic compounds (VOCs). Emissions from the ice resurfacers with combustion engines caused indoor air quality problems in all ice arenas. The highest 1-hour average CO and NO2 concentrations ranged from 20 to 33 mg/m3 (17 to 29 ppm) and 270 to 7440 micrograms/m3 (0.14 to 3.96 ppm), respectively. The 3 hour total VOC concentrations ranged from 150 to 1200 micrograms/m3. The highest CO and VOC levels were measured in the arena in which a gasoline-fueled resurfacer was used. The highest NO2 levels were measured in small ice arenas with propane-fueled ice resurfacers and insufficient ventilation. In these arenas, the indoor NO2 levels were about 100 times the levels measured in ambient outdoor air, and the highest 1-hour concentrations were about 20 times the national and World Health Organization (WHO) health-based air quality guidelines. The air quality was fully acceptable only in the arena with an electric resurfacer. The present study showed that the air quality problems of indoor ice arenas may vary with the fuel type of resurfacer and the volume and ventilation of arena building. It also confirmed that there are severe air quality problems in Finnish ice arenas similar to those previously described in North America. PMID- 9354146 TI - Nitrogen dioxide in indoor ice skating facilities: an international survey. AB - An international survey of nitrogen dioxide (NO2) levels inside indoor ice skating facilities was conducted. One-week average NO2 concentrations were measured inside and outside of 332 ice rinks located in nine countries. Each rink manager also completed a questionnaire describing the building, the resurfacing machines, and their use patterns. The (arithmetic) mean NO2 level for all rinks in the study was 228 ppb, with a range of 1-2,680 ppb, based on a sample collected at breathing height and adjacent to the ice surface. The mean of the second indoor sample (collected at a spectator's area) was 221 ppb, with a range of 1-3,175 ppb. The ratio of the indoor to outdoor NO2 concentrations was above 1 for 95% of the rinks sampled, indicating the presence of an indoor NO2 source (mean indoor:outdoor ratio = 20). Estimates of short-term NO2 concentrations indicated that as many as 40% of the sampled rinks would have exceeded the World Health Organization 1-hour guideline value of 213 ppb NO2 for indoor air. Statistically significant associations were observed between NO2 levels and the type of fuel used to power the resurfacer, the absence of a catalytic converter on a resurfacer, and the use of an ice edger. There were also indications that decreased use of mechanical ventilation, increased number of resurfacing operations per day, and smaller rink volumes were associated with increased NO2 levels. In rinks where the main resurfacer was powered by propane, the NO2 concentrations were higher than in those with gasoline-powered resurfacers, while the latter had NO2 concentrations higher than in those using diesel. Rinks where the main resurfacer was electric had the lowest indoor NO2 concentrations, similar to the levels measured outdoors. PMID- 9354147 TI - Meta-analyses of age-cognition relations in adulthood: estimates of linear and nonlinear age effects and structural models. AB - A meta-analysis was conducted on 91 studies to derive a correlation matrix for adult age, speed of processing, primary-working memory, episodic memory, reasoning, and spatial ability. Structural equation modeling with a single latent common cognitive factor showed that all cognitive measures shared substantial portions of age-related variance. A mediational model revealed that speed of processing and primary-working memory appear to be important mediators of age related differences in the other measures. However, not all of the age-related influences were mediated. An examination of quadratic age effects and correlational patterns for subsamples under and over 50 years of age revealed that (a) negative age-cognition relations were significant for the 18- to 50-year old sample and (b) the age-related decline accelerated significantly over the adult life span for variables assessing speed, reasoning, and episodic memory. PMID- 9354148 TI - Embarrassment: its distinct form and appeasement functions. AB - The authors address 2 questions about embarrassment. First, Is embarrassment a distinct emotion? The evidence indicates that the antecedents, experience, and display of embarrassment, and to a limited extent its autonomic physiology, are distinct from shame, guilt, and amusement and share the dynamic, temporal characteristics of emotion. Second, What are the theoretical accounts of embarrassment? Three accounts focus on the causes of embarrassment, positioning that it follows the loss of self-esteem, concern for others' evaluations, or absence of scripts to guide interactions. A fourth account focuses on the effects of the remedial actions of embarrassment, which correct preceding transgressions. A fifth account focuses on the functional parallels between embarrassment and nonhuman appeasement. The discussion focuses on unanswered questions about embarrassment. PMID- 9354149 TI - Finance. Healthy gains. PMID- 9354150 TI - Policy. Where there's smoke. PMID- 9354153 TI - Four costliest outpatient procedures. PMID- 9354151 TI - Philanthropy. How unbearably nice. PMID- 9354152 TI - Great comebacks. Divine results. Saved by sacrifice. PMID- 9354155 TI - Failed mergers. Trashed over local control. PMID- 9354154 TI - Why the phone company may be your best strategic partner. PMID- 9354157 TI - Pet therapy. Barking up the right tree. PMID- 9354156 TI - Faster CTs. What price innovation? PMID- 9354158 TI - Philanthropy. An uncharitable analysis. PMID- 9354159 TI - Online pharmacy. Rx for speedy service. PMID- 9354160 TI - The global settlement--a global view. PMID- 9354161 TI - The impact of residual disease on local recurrence in patients treated by initial unplanned resection for soft tissue sarcoma of the extremity. AB - BACKGROUND AND OBJECTIVES: "Unplanned excision" in soft tissue sarcoma (STS) is defined as the gross removal of tumor without preoperative staging or consideration of the need for removal of normal tissue around the tumor. This study evaluated whether unplanned excision in patients with extremity STS would have an impact on local relapse (LR), even if reexcision of the tumor bed was undertaken. METHOD: Two hundred thirty-nine (239) patients with primary, extremity STS treated with limb salvage surgery were included in this study. Of the 239, 78% were treated with surgery and irradiation. Forty-seven tumors were low-grade and 192 high grade. Mean tumor diameter was 7.5 cm. Twenty had local recurrences and 64 relapsed systemically. RESULTS: Only margin and prior surgery were significant in univariate analysis (P < 0.05, log-rank test). The Cox multivariate analysis revealed that both margin of resection (P < 0.001) and the status of the local tumor site (P < 0.05) at definitive surgery were significant predictors of local relapse. CONCLUSIONS: These results suggest that the presence of microscopic disease in the reexcised specimen following unplanned resection is a risk factor for local disease recurrence. PMID- 9354162 TI - Telomerase activity in esophageal carcinoma. AB - BACKGROUND AND OBJECTIVES: Telomerase is a ribonucleoprotein that synthesizes telomeric DNA. Immortalized and carcinoma cells show no loss of telomere length during cell division. Telomerase activity has been demonstrated in carcinomas of various organs, but not in nonneoplastic tissues. In patients with esophageal carcinoma, no data have been reported concerning the relationship between telomerase activity and clinicopathological findings. MATERIALS AND METHODS: Esophageal carcinomas from 31 patients and normal esophageal mucosae from 92 patients were examined. Telomeric Repeat Amplification Protocol assay to detect telomerase activity and Southern blot analysis to examine telomere length were performed. RESULTS: Of the 31 carcinomas, 27 (87%) had detectable telomerase activity. Twenty-one (23%) of the 92 normal esophageal mucosae from autopsied patients also had detectable telomerase activity. There was no difference between stage and outcome and absence or presence of telomerase activity. No difference in terminal restriction fragment (TRF) length was observed between carcinomas with and without telomerase activity. CONCLUSION: Telomerase activity was demonstrated in a considerable number of normal esophageal mucosae. This suggests the possibility of a high frequency of false positivity if the presence of telomerase activity alone is used as a tumor-specific marker. PMID- 9354163 TI - Massive bone allografts in the treatment of pathologic fractures due to bone metastases. AB - BACKGROUND AND OBJECTIVES: Pathologic fractures due to disseminated metastases are common and often involve major long bones, where the metastasis is responsible for wide bone erosion that is equivalent to major bone loss. Stabilization of these fractures necessitates tumor excision and reconstruction of the destructive metastatic process. METHODS: Massive allografts were used either as intercalary or "composite" grafts (allografts and regular prostheses) in 17 patients. RESULTS: Fourteen patients could ambulate independently after surgery, and nursing of the remaining three became feasible and less painful. CONCLUSION: In the event of pathologic fractures due to metastatic bone diseases associated with major bone involvement, massive bone allografts offer an inexpensive, adjustable, simple, solution. PMID- 9354164 TI - K-ras gene mutations in intrahepatic bile duct tumors of Syrian golden hamsters. AB - BACKGROUND AND OBJECTIVES: In our laboratory, we have developed a new model of carcinoma of the bile duct in Syrian golden hamsters, using N-nitrosobis(2 oxopropyl)amine (BOP). Morphologic and biologic characteristics of the carcinoma induced in this model are similar to those seen in humans. In order to examine the gene-related carcinogenesis of intrahepatic bile duct carcinoma, we investigated mutations in the K-ras gene in various early hyperplastic and neoplastic lesions of these hamsters, according to the original sites of the lesions. METHODS: Inbred female hamsters were given a subcutaneous injection of N nitrosobis(2-oxopropyl)amine (BOP) following dissection of the extrahepatic bile duct on the distal end of the common duct and preparation of a cholecystoduodenostomy (CDDB) or simple laparotomy (SL). Neoplastic lesions arising from the intrahepatic bile duct were histologically examined, and K-ras mutations were investigated. RESULTS: Mutations of K-ras codon 12 were evident in 12% of tubular hyperplasias, 19% of tubular adenocarcinomas, 15% of papillary hyperplasias and 36% of papillary adenocarcinomas. In papillary adenocarcinoma arising from a large bile duct, K-ras mutations occurred more frequently than in tubular adenocarcinoma arising from ductule or ductular proliferation. K-ras mutations were present even in a hyperplasia; the positive rates of the mutations increased in the presence of a carcinoma. Genetic changes in carcinoma of the intrahepatic bile duct varied based on sites of the duct and the histological type. CONCLUSIONS: A part of the hyperplastic lesions of the intrahepatic bile duct presented K-ras gene mutation. This suggests that K-ras gene mutation is an early event in the carcinogenic process. In carcinoma of the intrahepatic bile duct, the lesion arising from a large bile duct of the hepatic hilum tended to exhibit a higher frequency of K-ras gene mutation than did a tubular lesion arising from ductule or ductal proliferation. This hamster model is useful to examine the carcinogenesis of human intrahepatic bile duct carcinoma. PMID- 9354165 TI - Tumor bed brachytherapy with a mesh template: an accessible alternative to intraoperative radiotherapy. AB - BACKGROUND AND OBJECTIVES: Locally advanced and recurrent malignancies often require adjuvant radiotherapy to achieve tumor control. We report our experience with a technique that uses an intraoperatively placed mesh template for the delivery of radiotherapy. METHODS: from 1988 to 1996, 14 patients were treated with tumor bed brachytherapy using this mesh technique. Sites of involvement included the head and neck region (n = 6), abdomen/pelvis (n = 4), retroperitoneum (n = 3), and the lower extremity (n = 1). During surgery, plastic catheters were evenly placed within a mesh template (Vicryl or Marlex), which was positioned in the tumor bed. The catheters were afterloaded with radioactive sources once the final pathology had been determined and the patient required limited nursing care. Radiation dose was titrated to the surgico-pathologic findings (e.g., margin status). RESULTS: All of the patients tolerated the procedure without experiencing acute or chronic sequelae. The median survival time was 13 months. Local control was achieved in 11 of 13 evaluable patients, with an actuarial local control of 82% at 6 months. CONCLUSION: Tumor bed brachytherapy with a mesh implant is a practical technique to improve tumor control and warrants further investigation. PMID- 9354166 TI - Use of a moratorium to achieve consistent liquid nitrogen cryoprobe performance. AB - BACKGROUND AND OBJECTIVES: During liver cryosurgical procedures, the authors observed seriously inconsistent rates of iceball growth implying inconsistent probe cooling rates. This inconsistency can lead to several problems, most importantly, loss of precision and reliability in freezing the chosen volume of tissue. The observation led to investigation of the performance of the cryosurgery machine. METHODS: The performance of the Cryotech LCS 3000 with different moratorium periods was investigated. Freezing was performed in a gelatin phantom with thermocouple monitoring at the probe tip. RESULTS: It was determined that a moratorium period between unit filling and clinical use essentially eliminates the uncertainty in probe performance. The need for this arises from the design of the system. A full liquid nitrogen tank does not have the necessary pressure required to induce effective freezing. A partially filled tank increases vapor pressure in the storage dewar driving more liquid nitrogen through the probes, thus allowing a more reliable and faster freeze. CONCLUSION: The simple measure of introducing a moratorium period between filling the dewar with liquid nitrogen and its use in surgery allows partial evaporation of the nitrogen and enhances cryoprobe performance. This protocol modification may reduce the chance of inconsistent probe performance thus making liver cryosurgery more reliable. PMID- 9354167 TI - Chemical adjuvant cryosurgery with antifreeze proteins. AB - BACKGROUND AND OBJECTIVES: Imaging monitored cryosurgery is emerging as an important minimally invasive surgical technique for treatment of cancer. Although imaging allows excellent control over the process of freezing itself, recent studies show that at high subzero temperatures cells survive freezing. Antifreeze proteins (AFP) are chemical compounds that modify ice crystals to needle-like shapes that can destroy cells in cellular suspensions. The goal of this study was to determine whether these antifreeze proteins can also destroy cells in frozen tissue and serve as chemical adjuvants to cryosurgery. METHODS: Livers from six rats were excised, perfused with solutions of either phosphate-buffered saline (PBS) or PBS with 10 mg/ml AFP-I, and frozen with a special cryosurgery apparatus. Lobes were frozen with one or two freeze-thaw cycles and the cell viability was examined with a two stain fluorescent dye test and histological assessment. RESULTS: A significant percentage of hepatocytes survive freezing on the margin of a frozen cryolesion. AFP significantly increase cellular destruction in that region apparently through formation of intracellular ice. CONCLUSIONS: This preliminary study demonstrates that antifreeze proteins may be effective chemical adjuvants to cryosurgery. PMID- 9354168 TI - Prognostic significance of surgical margin in hepatocellular carcinoma resection: an analysis of 165 Childs' A patients. AB - BACKGROUND AND OBJECTIVES: The clinical significance of the width of the surgical margin in the resection of hepatocellular carcinoma (HCC) has yet to be clarified. METHODS: Childs' A patients (165) who underwent resections of HCC were studied. Patients were divided into a wide margin group (1.0 cm or more, group W, n = 85), and a narrow margin group (< 1.0 cm, group N, n = 80). RESULTS: Multivariate analysis showed that preoperative alpha-fetoprotein level (P = 0.0202), venous invasion (P = 0.0226), surgical margin (P = 0.0012), and TNM stage (P = 0.0023) were significant predictors of disease-free survival. By the log-rank test, the disease-free survival rate of the group W patients was significantly higher than that of the group N patients (P = 0.0007). Group N had a higher percentage of patients undergoing minor resection (wedge resection or subsegmentectomy) (44% vs. 26%, P = 0.016) and had a higher percentage of patients with centrally located tumor (62% vs. 29%, P = 0.000) than group W. CONCLUSIONS: The results of this study indicated the significant influence of surgical margin on HCC recurrence after resection. Minor resection and centrally located tumor are factors related to a narrow surgical margin. PMID- 9354169 TI - Esophagojejunostomy with manual single layer suturing after a total gastrectomy for gastric cancer. AB - BACKGROUND: We wished to verify the clinical usefulness of manually performed single layer suturing for an esophagojejunostomy after a total gastrectomy versus stapled suturing. METHODS: We compared retrospectively 24 patients who underwent manual single layer suturing with 38 patients who underwent stapled suturing. RESULTS: Anastomotic leakage was seen in one patient (4%) with single layer suturing and one patient (3%) with stapled suturing. No anastomotic stenosis was seen in the patients with single layer suturing. There was no difference in the operative time, blood loss, postoperative days for oral intake, or the length of hospital stay between the patients with single layer suturing and those with stapled suturing. CONCLUSIONS: Manual single layer suturing is considered to be as safe as stapled suturing and is also thought to be clinically useful in reducing anastomotic failure for esophagojejunostomy. PMID- 9354170 TI - Second primary carcinoma in the residual cervical esophagus after thoracic esophagectomy: report of five cases. AB - BACKGROUND AND OBJECTIVES: Development of second primary carcinomas after thoracic esophagectomy has become of much concern, because recently the prognosis of thoracic esophageal carcinoma after esophagectomy with extended lymph node dissection has been improving. We report our experience of diagnosing and treatment second primary carcinomas arising in the remaining esophagus after thoracic esophagectomy. METHODS: Among 253 patients who underwent esophagectomy for thoracic esophageal carcinoma more than 2 years previously, second primary esophageal carcinomas developed in five (2.0%), and these five patients were examined. RESULTS: All second primary carcinomas were found by endoscopy, and were diagnosed as superficial carcinoma (Tis or T1) of the residual cervical esophagus. One patient underwent laser irradiation, another endoscopic mucosal resection, two had surgical mucosectomy, and one segmental resection of the esophagus. After the second treatment, three patients were disease free for 37-38 months, one died of recurrent disease of the first carcinoma 36 months later, and one died of distant metastases of the second carcinoma 8 months later. There have been no local recurrences after treatments for the second primaries. CONCLUSIONS: A variety of low-trauma treatments were employed for the second carcinomas because they were found at an early stage. Endoscopic follow-up is proposed to detect second lesions at an early stage. PMID- 9354172 TI - Palliative surgical treatment in enterocutaneous fistula. PMID- 9354171 TI - Repair of tracheal defect with Goretex graft during resection of carcinoma of the esophagus. AB - Repair options for tracheal defects secondary to tumor or trauma have been unsatisfactory for emergent cases. We report a case in which the tracheobronchial tree was entered during resection of carcinoma of the esophagus and emergently repaired with a Goretex graft. The patient did well for 22 months after esophagectomy, at which time the graft was found to be infected and was removed. The patient continues to remain free of tumor 4 years after initial resection. PMID- 9354173 TI - Intra-abdominal side-to-end stapling technique for anterior resection of rectal cancer. PMID- 9354174 TI - Intracavitary brachytherapy in the treatment of gynecologic neoplasms. AB - Modern intracavitary brachytherapy carefully combined with megavoltage external beam radiotherapy is responsible for the high cure rates achieved with radiation treatment of invasive cervical cancers. Pelvic disease recurrence is rare after treatment of patients with tumors < 5 cm in diameter, and even patients with massive tumors 8-10 cm in diameter are cured in 30-50% of cases. Inoperable adenocarcinomas of the endometrium and superficial cancers of the vagina are also effectively treated with intracavitary irradiation. The relative radioresistance of the uterus and vagina, physical advantages resulting from exploitation of the inverse square law, and the radiobiological advantages of low dose rate radiation have combined to make intracavitary irradiation a critical tool in the management of many gynecologic neoplasms. PMID- 9354175 TI - Comparison of organochlorine pesticide and polychlorinated biphenyl residues in human breast adipose tissue and serum. AB - The presence of organochlorine pesticides, such as p,p'-DDT[2,2-bis(p chlorophenyl)-1,1,1-trichloroethanel, and of polychlorinated biphenyls (PCBs) in human serum and adipose tissue has been reported in many studies over the last four decades. Recently, debate has heightened concerning the link of these compounds to breast cancer. To clarify and resolve this issue, accurate analytical residue data must be obtained. Separation of the organochlorine pesticides from the PCBs in breast tissue is critical to obtaining valid residue data. Based on methods refined in the Analytical Laboratory at Colorado State University, accurate residue levels were established for nine individual PCB congeners and eight organochlorine pesticides. The breast adipose tissue method used was a modification of the Mills et al. and de Faubert Maunder et al. methods. The serum method employed was a modification of the Burse et al. method. Both breast adipose tissue and serum from 36 women were analyzed, and correlations of the residues from the two substrates were evaluated. Serum concentrations of p,p'-DDE, the primary metabolite of p,p'-DDT, were correlated (alpha = .05) with the concentrations of p,p'-DDE in human breast adipose tissue (r = .808). Serum concentrations of the PCB congener BZ 153 were also significantly correlated to the human breast adipose tissue concentrations of BZ 153 (r = .377). No significant relationship was found between serum concentrations and tissue residues for 15 of the 17 compounds analyzed. This lack of correlation between breast adipose tissue and serum, as well as an absence of the compound residues in serum, emphasized that adipose tissue should be analyzed in addition to serum to fully understand the relationship of the organochlorine compounds to breast cancer. PMID- 9354176 TI - Estimating pesticide exposure in tidal streams of Leadenwah Creek, South Carolina. AB - This article estimates the potential exposure of estuarine organisms to two pesticides (azinphosmethyl and fenvalerate) in a tidal stream of Leadenwah Creek near the Edisto River, South Carolina, during four runoff episodes. Exposure is calculated from simulation runs of the one-dimensional transport equation solved by an implicit finite difference method. Calibration was done for each episode by adjusting three conditions (runoff starting time, duration, and flow) and a correction to the dispersion coefficient in order to match the continuously measured salinity transients. First-order rate constants used by the fate component were calculated from half-life values reported in the literature. Baseline scenarios for each episode and each pesticide were derived by using the same conditions obtained in the salinity runs and adjusting the pesticide loading in order to mimic the few data points of measured pesticide concentrations. In all baseline scenarios, pesticide concentration rises following the initial burst of runoff (also noticeable as an abrupt drop in salinity) and then oscillates, forced by the tidal cycle. These oscillations are dominated by transport, while fate imposes a secular decaying trend. Ten additional scenarios for each episode were obtained from the baseline scenario by randomly varying three pesticide load parameters (starting time and duration of runoff, and pesticide discharge) using a Latin hypercubes design. Two exposure metrics were calculated from the simulated and the measured pesticide concentration: maximum and time average, which was obtained by integrating the curve and dividing by the time period. The metrics calculated from the baseline runs are relatively close to the data derived metrics, because the baseline runs attempted to mimic the data. For each one of the two metrics and all pesticide-episode combinations, several statistics of the set of 11 scenarios were also calculated: minimum and maximum, mid-range, mean, standard deviation, and median. The mean +/- standard deviation interval of the simulation-derived value consistently brackets the data-derived value for the maximum metric, but not for the time-average metric. This may indicate that even if the maximum value is correctly captured in the field sample, the time-average exposure could be in error when calculated directly from the field data due to undersampling of the pesticide time series. The methodology developed here attempts to reconstruct the possible exposure from the sparse sampling of the pesticide concentration during the runoff episodes; only when the number of field samples is high and regularly spaced is it possible to have confidence in the reconstruction of the curve. The shape of the curve cannot be inferred from the field measurements alone; as expected, tidal movement makes the pesticide concentration swing up and down. This result has important implications because the biological community would be subject to repetitive pulses of exposure to the chemicals. The baseline simulations can be used to derive a pulse-exposure metric by calculating the sum of ratios of the time average of the threshold-exceeding concentrations to the time average of the toxic threshold during intervals of above-threshold concentration. This metric is species specific and extrapolates laboratory toxicity data in order to compare pulse exposure to mortality rates measured in the field. PMID- 9354177 TI - Correspondence of salivary and plasma concentrations of atrazine in rats under variable salivary flow rate and plasma concentration. AB - The stability of the saliva/plasma (S/P) concentration ratio of atrazine was determined under varying conditions of salivary flow rate and plasma concentration of atrazine in Sprague-Dawley rats. In the salivary flow study, whole saliva samples were collected at different salivary flow rates while the plasma concentration of atrazine was maintained at a steady-state level of approximately 150 micrograms/L. In the plasma level study, whole saliva samples were collected at two steady-state plasma concentrations of atrazine (50 and 250 micrograms/L), while salivary flow rate was maintained at a relatively constant level. In both studies, atrazine concentrations in whole saliva and arterial plasma demonstrated a consistent relationship, but salivary concentrations were always lower than those of arterial plasma. Linear regression analysis demonstrated that the S/P concentration ratio of atrazine was not significantly different for salivary flow rates ranging from 23 to 92 microL/min/kg body weight, and did not vary for atrazine plasma concentrations between 30 and 433 micrograms/L. The S/P concentration ratio of atrazine was relatively constant throughout each experimental period (0.68 +/- 0.1 and 0.70 +/- 0.11 for salivary flow and plasma level studies, respectively) and did not differ significantly between rats. When data from both studies were pooled, salivary concentrations were highly correlated with plasma concentrations (r2 = .94). It is concluded that under these experimental conditions, the stability of the S/P concentration ratio of atrazine is not affected by variations in salivary flow rate or atrazine plasma concentrations. The results from this study support the conclusion that atrazine salivary concentrations can be used to predict plasma levels of atrazine in rats. PMID- 9354178 TI - Metabolism of arsenic after acute occupational arsine intoxication. AB - Among the elements of toxicological relevance, inorganic arsenic (As) probably exhibits the most complex metabolism, and we deemed it interesting to identify and quantify the different As species excreted after an occupational acute intoxication with arsine. For this purpose total As and five As species were determined using an hybrid analytical method coupling liquid chromatography with inductively coupled plasma mass spectrometry. The highest urinary elimination of total As was observed in the first 5 d after admission. The As species mostly excreted were monomethylarsonate (MMA), dimethylarsinate (DMA), As3+, arsenobetaine (AsB), and to a lesser extent As5+. The amount of AsB excreted in urine by the subject does not appear to be completely justified by AsB intake through food. Arsenic is excreted mainly via the urine with a clearance of 7.8 ml/h/kg and follows a triphasic model with periods of 28 h, 59 h, and 9 d, respectively. The evidence that DMA excretion culminates after a few days, when the excretion of the inorganic form is substantially reduced (while that of MMA is still elevated), seems to confirm the existence of two successive methylating enzyme activities. Furthermore, the elimination rate of As from blood follows a three-phase model and the half-lives of different species vary from about 27 to 86 h with the following gradient As5+ < MMA < As3+ < DMA < AsB. PMID- 9354179 TI - Ferritin adsorption on amosite fibers: possible implications in the formation and toxicity of asbestos bodies. AB - In order to investigate how endogenous iron can be deposited in vivo on inhaled mineral fibers during early stages of formation of asbestos bodies, in vitro experiments were performed on the adsorption of ferritin onto amosite asbestos. The mineral dust was found to adsorb the protein from an aqueous solution containing 0.3 mg/ml horse spleen ferritin. In order to simulate physiological conditions the aqueous solution was adjusted with 150 mM saline. Polyacrylamide SDS gel electrophoresis of the desorbed protein showed subunits of approximately 13 and 15 kD, aside from the 20-kD subunit present in the native protein. This suggests that as a result of interactions between ferritin molecules and the solid surface of the mineral fibers, the protein iron core may be released or partially exposed. Data indicate these interactions may have implications in the observed mineral fiber toxicities. PMID- 9354181 TI - Effect of pretreatment with dichloroacetate or trichloroacetate on the metabolism of bromodichloroacetate. AB - Haloacetates are a common class of water chlorination by-products. Depending on the amount of bromide in the source water, varying amounts of chlorinated, brominated, and mixed bromochloro haloacetates are produced. When administered to rodents, haloacetates have been shown to increase formation of thiobarbituric acid-reactive substances and 8-hydroxydeoxyguanosine levels in the liver. These responses appear to be modified by prior treatment. To examine potential mechanisms that account for these modifications in oxidative stress, the ability of trichloroacetate (TCA) or dichloroacetate (DCA) pretreatment to alter the metabolism of bromodichloroacetate (BDCA) and the disposition of its metabolites was examined in male B6C3F1 mice. Two-week pretreatment with 1 g/L DCA and TCA in the drinking water of mice alters the initial hepatic metabolism of BDCA and the further metabolism of its metabolite DCA. DCA pretreatment inhibits cytosolic metabolism of both 1 mM DCA or BDCA up to 70%. In contrast, DCA pretreatment stimulates hepatic microsomal BDCA metabolism 1.3-fold but has little effect on microsomal metabolism of DCA. Increased microsomal metabolism of BDCA appears to be attributable to the induction of a metabolic pathway that produces CO2 and bromodichloromethane (BDCM) as metabolites. TCA pretreatment inhibits BDCA metabolism up to 70% in the cytosol and 30% in microsomes but has little effect on DCA metabolism. These results indicate that the hepatic metabolism of the haloacetate becomes quite complex at the high doses that have been employed in cancer bioassays. BDCA serves as a good example, because it is metabolized to at least two carcinogenic metabolites that have different modes of action, BDCM and DCA. As doses approach those that induce cancer in mice, the proportion of and amounts of these metabolites as a fraction of the dose administered will change substantially. This article demonstrates that those interactions will occur from mixed treatment with haloacetates as well. PMID- 9354182 TI - Age-associated endocrine deficiencies as potential determinants of femoral neck (type II) osteoporotic fracture occurrence in elderly men. AB - Osteoporotic fractures, and especially hip fractures, are a leading cause of morbidity and mortality among elderly men. Among other factors, a decline in bone mass has been identified as the major determinant of the age-related reduction in bone strength and therefore of osteoporotic fracture risk. Recent evidence suggests that age-associated endocrine deficiencies may contribute to femoral bone loss and hip fracture occurrence in elderly men. The decline in circulating androgen levels and the decreased activity of the growth hormone insulin-like growth factor-I axis may result in a reduction in bone formation that contributes to the age-related increase in bone fragility in men. Vitamin D deficiency induced secondary hyperparathyroidism, on the other band, may further enhance bone loss by activating bone turnover and so increasing the number of bone remodelling units with impaired bone formation. On the basis of these pathophysiological models, guidelines can be developed for the prevention of age related bone loss in men, but these approaches lack validation. The results of controlled intervention trials will have to be awaited to answer the question of whether hormone replacement therapy attenuates bone loss and reduces fracture incidence in elderly men. PMID- 9354180 TI - Amiodarone-induced pulmonary toxicity in Fischer rats: release of tumor necrosis factor alpha and transforming growth factor beta by pulmonary alveolar macrophages. AB - Amiodarone is an antiarrhythmic drug with numerous side effects, the most serious being the development of pulmonary toxicity. We have previously reported that a single intratracheal instillation of amiodarone to Fischer 344 rats results in pulmonary fibrosis within 6 wk of treatment. Presently, the mechanism of amiodarone-induced pulmonary toxicity is unknown. Cytokines that stimulate fibroblast proliferation and/or collagen production may play a role in amiodarone induced pulmonary toxicity. To investigate this possibility, female rats were given a single intratracheal instillation of amiodarone (6.25 mg/kg), its metabolite desethylamiodarone (5 mg/kg), or vehicle (sterile water). At 1, 2, 3, or 6 wk after treatment the lungs were lavaged and the recovered cells were counted and identified. The alveolar macrophages were isolated by attachment to plastic petri dishes, cultured overnight, and the spent media collected for tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) analyses. Desethylamiodarone treatment resulted in a neutrophilic alveolitis, but the levels of TNF-alpha and TGF-beta were not significantly different from control animals. In contrast, amiodarone treatment resulted in a lymphocytic alveolitis and significantly higher amounts of TNF-alpha were observed at 3 and 6 wk after treatment. A trend toward higher levels of TGF-beta was also noted in the amiodarone-treated group at wk 1-3 but the values were not significantly different from those of controls. In conclusion, the release of TNF-alpha may play a role in the development of amiodarone-induced pulmonary toxicity. PMID- 9354183 TI - Frequently subnormal semen profiles of normal volunteers recruited over 17 years. AB - In order to establish semen profiles for men in the general population, we analysed the semen parameters of 187 men attending the Institute of Reproductive Medicine between 1977 and 1993 as volunteers for clinical studies. If several ejaculates were obtained from the volunteer, only the first ejaculate was used for analysis. More than half of the ejaculates of these healthy men showed at least one abnormal parameter, so that only 46% of the volunteers could be classified as being 'normozoospermic' according to WHO guidelines. Aside from a decrease in the proportion of motile spermatozoa, no change in semen parameters could be detected with advancing age of the volunteers. No negative influence of the presence of a varicocele, the grade of varicocele on a history of maldescended testis on semen parameters was detected. The serum levels of FSH, LH and testosterone showed no significant age-related change over the period of investigation. Follow-up analysis of the fertility on the volunteers revealed a proportion of unwanted barrenness similar to or even lower than that in the general population. The analysis demonstrates that the limits of normality for semen parameters may require redefinition. PMID- 9354184 TI - Pituitary, gonadal and adrenal hormones after prolonged residence at extreme altitude in man. AB - High altitude-induced alterations in pituitary, gonadal and adrenal hormones were studied in (i) eugonadal men from the armed forces who were resident at sea level (SL), (ii) SL residents staying at an altitude of 3542 m for periods ranging from 3 to 12 months (acclimatized lowlanders, ALL), (iii) ALL who stayed at 6300 m for 6 months, (iv) ALL who trekked from 3542 to 5080 m and stayed at an altitude of more than 6300 m in the glacier region for 6 months, and (v) high-altitude natives (HAN) resident at an altitude of 3300-3700 m. Circulating levels of LH, FSH, prolactin, cortisol, testosterone, dihydrotestosterone (DHT) and progesterone in ALL at 3542 m and in HAN were not significantly different (p > 0.05) from the SL control values. When the ALL living at 3542 m trekked to an extreme altitude of 5080 m, their testosterone levels showed a significant decrease (p < 0.01) compared to the preceding altitude values but had returned to SL values when measured after 6 months' continuous stay at 6300 m. As with testosterone, the levels of DHT and oestradiol-17 beta (E2) after prolonged stay at extreme altitude were also not significantly different (p > 0.05) from the SL values. The LH levels after trekking to 5080 m were significantly higher (p < 0.01) than at an altitude of 3542 m, but decreased to levels found at 3542 m or SL after prolonged residence at extreme altitude. Plasma levels of ACTH, prolactin, FSH and cortisol on arrival at 5080 m, and after a 6-month stay at extreme altitude, were not significantly different (p > 0.05) from the SL values. Plasma progesterone levels tended to increase on arrival at 5080 m but a significant increase (p < 0.001) was evident only after a 6-month stay at extreme altitude. These observations suggest that prolonged residence at lower as well as at extreme altitude does not appreciably alter blood levels of pituitary, gonadal or adrenal hormones except for plasma levels of progesterone. The exact mechanism and significance of this increase remains unknown, but may be important in increasing the sensitivity of the hypoxic ventilatory response and activation of haemoglobin synthesis. PMID- 9354185 TI - Prevalence and risk factors for anabolic-androgenic steroid abuse in Australian high school students. AB - Abuse of anabolic-androgenic steroids (AAS) is reportedly increasing in prevalence and spreading from sportsmen to cosmetic and recreational use, though there are few accurate estimates of the extent of AAS use outside North America. In order to identify the prevalence of, and risk factors for, 'ever' (lifetime) or 'recent' (within last month) use of AAS among Australian high school students, 13,355 students (51.3% male, median age 13.8 years) in 203 schools constituting a stratified random sample of all high schools in New South Wales and Victoria completed a self-report questionnaire about drug use. Lifetime ('ever') use of AAS was reported by 213 boys (3.2%; 95% CI, 2.7-3.6%) and 74 girls (1.2%; 95% CI, 0.9-1.5%). Factors associated with 'ever' use were truancy [odds ratio (OR) 17.7 (95% CI, 9.4-31.6) for 7 or more truant days compared with no trauncy over the last fortnight], unsupervised recreation [OR 8.4 (5.9-11.9) for 6 or more compared with 2 or fewer nights per week], speaking only a non-English language at home [OR 7.75 (4.4-13.1)], Aboriginality [OR 3.4 (2.0-5.5)], male gender [OR 2.8 (2.1-3.7)], higher student income [OR 2.3 (1.7-3.0)], overseas born [OR 2.2 (1.6-3.0)] and low level of social support [OR 2.5 (1.7-3.5)]. Use of AAS within the last month ('recent') was reported by 113 boys (1.7%; 95% CI, 1.4-2.0%) and 28 girls (0.4%; 95% CI, 0.3-0.6%) and virtually identical risk factor patterns except ORs were uniformly higher. We conclude that AAS abuse is relatively uncommon among Australian high school students but has distinctive and strong socio-economic and cultural predictors. Further studies of the behavioural epidemiology of AAS abuse are required to clarify the origins and significance of our findings and to identify potentially effective interventions. PMID- 9354186 TI - Evidence that peptides derived from the disintegrin domain of primate fertilin and containing the ECD motif block the binding of human spermatozoa to the zona free hamster oocyte. AB - The binding of human spermatozoa to, and penetration of, zona-free hamster oocytes were significantly inhibited in the presence of (i) a 28-residue peptide corresponding to part of the disintegrin-like domain of monkey fertilin beta, or (ii) a hexapeptide containing the ECD motif found in the disintegrin-like domain of monkey and human fertilin beta, or (iii) a hexapeptide containing an RGD motif, implicated in integrin recognition. Polyclonal antibodies raised against the 28-residue peptide were used in conjunction with immunobeads to demonstrate that this peptide bound to the oolemma of zona-free hamster oocytes. These data support the view that the ECD motif is involved in the recognition of the oolemma receptor by human fertilin. PMID- 9354187 TI - Regulation of the secretion and synthesis of rat Sertoli cell SGP-1, SGP-2 and CP 2 by elongate spermatids. AB - The aim of this study was to investigate the effect of the absence of elongate spermatids (ES) from the rat seminiferous epithelium on the quantitative secretion and synthesis of the three major Sertoli cell secretory proteins--SGP 1, SGP-2 and CP-2. Seminiferous tubules (ST) were isolated (a) from normal 28-day old rats, in which the most mature germ cell type is the round spermatid, (b) from normal adult rats at stages IX-XIV of the spermatogenic cycle, i.e. after spermiation, or at stages I-V and VI-VIII, when ES are still attached to the Sertoli cell, and (c) at stages VI-VIII from normal adult rats and from rats treated with methoxyacetic acid (MAA) in order to specifically deplete ES at these stages. Two-dimensional SDS PAGE combined with computerized image analysis was used to analyse 35S-methionine-labelled intracellular and secreted proteins. In the case of SGP-1 and SGP-2, almost all of the protein synthesized by ST was secreted. The total amount of both SGP-1 and CP-2 secreted by unstaged ST from immature rats was significantly lower than that secreted by unstaged ST from adult rats. The total amount of SGP-1 and CP-2 secreted by adult ST at stages IX XIV of the spermatogenic cycle also declined dramatically compared to ST at earlier stages. The proportion of the total CP-2 synthesized by ST which was secreted also declined in all situations in which ES were absent from the seminiferous epithelium. The synthesis of only SGP-2 was changed by ES depletion from ST at stages VI-VIII, which was almost doubled compared to synthesis of this protein by ST from control rats. Our results suggest strongly that the secretion of SGP-1 and SGP-2 is via the constitutive pathway, and that regulation of these two proteins by ES is at the level of protein synthesis. In contrast, the regulation of CP-2 by ES is predominantly at the level of secretion, suggesting that this protein is secreted via a regulated pathway. Our findings add to the evidence showing that ES play a major role in the regulation of Sertoli cell function. PMID- 9354188 TI - Successful lowering of epididymal carnitine by administration of pivalate to rats. AB - This study was designed to lower the epididymal content of carnitine in male rats and to examine subsequent effects on fertility and sperm motility. As carnitine is transported from serum into the epididymal lumen a method to lower serum carnitine was sought. Administration of 20 mmol/L sodium pivalate in the drinking water for up to 5 weeks substantially lowered serum carnitine (to 20% of control levels within 1 week) and reduced epididymal carnitine content (to 25% of control levels in the proximal and 52% of control in distal regions) within 2 weeks. Carnitine in distal cauda epididymal fluid was also reduced (to 30% of control levels) but no changes were observed in the sperm carnitine content. The percentage motility and kinematic parameters of spermatozoa released from four epididymal regions and diluted into artificial medium were unaltered by the treatment, and all males retained their fertility in mating tests performed at weekly intervals. Increasing the dose of sodium pivalate administered to 60 mmol/L for 2 weeks lowered serum carnitine concentration more but did not further decrease epididymal carnitine content and altered neither sperm motility nor male fertility. The rat epididymis secretes an excessive amount of carnitine into its lumen so that substantially lowering the tissue content does not reduce sperm carnitine or affect their motility or fertilizing ability. PMID- 9354189 TI - Review article: gene therapy in gastroenterology and hepatology. AB - Gene therapy for diseases of the gastrointestinal tract is an exciting prospect because of the fundamental cure that is potentially available. The gastrointestinal system, and especially the liver, is an area that will be central to the development of gene therapy. Techniques for gene replacement include homologous recombination and gene augmentation. For the treatment of cancer antisense strategy, pro-drug activation systems and gene immunotherapy are being investigated. Gene-carrying vectors divide into viral- and non-viral-based vectors, each with advantages and limitations. The accurate delivery of these vectors to sufficient numbers of target cells in vivo is still a major barrier to clinical use. Diseases that may be helped by gene therapy include: gastrointestinal malignancies, viral hepatitis, the haemophilias, hypercholesterolaemia, alpha 1-antitrypsin deficiency, and metabolic diseases of the liver and cystic fibrosis. In this review we will outline the principles of gene therapy, delivery vectors under investigation, diseases that may benefit from this technology and some of the remaining problems to be overcome. PMID- 9354190 TI - Effects of oral cisapride on small bowel motility in irritable bowel syndrome. AB - BACKGROUND: Cisapride has been reported to improve symptoms in patients with constipation-predominant irritable bowel syndrome. AIM: To compare the effects of a 24-h oral dose regimen of cisapride on interdigestive and post-prandial small bowel motor activity in irritable bowel syndrome patients with predominant constipation, irritable bowel syndrome patients with predominant diarrhoea and healthy subjects. METHODS: In 12 irritable bowel syndrome patients (11 females, aged 44 +/- 12 years)--constipation-predominant (irritable bowel syndrome-C, n = 5) and diarrhoea-predominant (irritable bowel syndrome-D, n = 7)--and six healthy subjects, small bowel motor activity was continuously recorded using an ambulatory technique over a 48-h period. Subjects received, in single-blind fashion, placebo tablets q.d.s. in the first 24 h then cisapride 10 mg q.d.s. in the second 24 h. Additional control groups were 13 healthy subjects (eight females, aged 39 +/- 13 years) and 10 irritable bowel syndrome patients (10 females, aged 49 +/- 14 years) who were studied in identical fashion but who did not receive cisapride. RESULTS: Cisapride increased migrating motor complex phase 2 motility index in both irritable bowel syndrome-D (P < 0.01) and irritable bowel syndrome-C (P < 0.05) patients, as well as in healthy subjects (P < 0.01). An increase in fasting discrete clustered contractions occurred in irritable bowel syndrome-D patients (P < 0.001) and in healthy subjects (P < 0.01), but not in irritable bowel syndrome-C patients; the proportion of discrete clustered contractions that were propagated, however, increased only in irritable bowel syndrome-D patients (P < 0.001). In addition, cisapride resulted in an increase in post-prandial motility index in irritable bowel syndrome patients (P < 0.05). Such motor alterations were not observed during the 48-h recording period in the healthy or irritable bowel syndrome patient control groups who did not receive cisapride. CONCLUSIONS: Oral cisapride influences interdigestive and post prandial small bowel motor activity in both irritable bowel syndrome patients and healthy subjects; the effects of cisapride may be more marked in patients with predominant diarrhoea than in patients with predominant constipation. PMID- 9354191 TI - Controlled trial of oral 5-aminosalicylic acid for the prevention of early relapse in Crohn's disease. AB - BACKGROUND: Recent data indicate that 5-aminosalicylic acid (5-ASA) is most effective in preventing relapse of Crohn's disease in patients with a short duration of remission before enrollment. AIM: To evaluate the efficacy of oral 5 ASA treatment, started immediately after achieving steroid-induced remission, in preventing clinical relapses of Crohn's disease. METHODS: Patients with active Crohn's disease, achieving remission on steroids, were randomized to oral 5-ASA 3 g/day or placebo, while steroids were tapered over 6 weeks. The trial was terminated after interim analysis showed a slightly higher relapse rate in the 5 ASA group, and the calculated probability of seeing a statistically significant difference by completing the study was minimal. RESULTS: Final analysis included 117 patients (58 taking 5-ASA and 59 taking placebo; follow-up 9.2 +/- 6.5 months). Cumulative relapse rates at 6 and 12 months were 34% and 58% in 5-ASA patients and 31% and 52% in placebo patients, respectively (rate difference +0.095; 95% CI = -0.085 to +0.274). Subgroups analysis showed that 5-ASA was equally ineffective in patients with ileal, colonic or ileocolonic disease. CONCLUSIONS: Contrary to previous results, in our study early introduction of treatment with oral 5-ASA did not prevent relapse in Crohn's disease patients treated with steroids to induce remission. PMID- 9354192 TI - Double-blind comparison of an oral Escherichia coli preparation and mesalazine in maintaining remission of ulcerative colitis. AB - BACKGROUND: Aminosalicylates are used as standard treatment for maintaining remission in ulcerative colitis. As yet, there is no other existing alternative with proven efficacy. In light of the hypothesis that the intestinal environment may contribute to the pathophysiology of ulcerative colitis, a trial was conducted to test the effects of probiotic treatment with an oral preparation of non-pathogenic E. coli. METHODS: A total of 120 patients with inactive ulcerative colitis were included in a double-blind, double-dummy study comparing mesalazine 500 mg t.d.s. to an oral preparation of viable E. coli strain Nissle (Serotype 06: K5: H1) for 12 weeks with regard to their efficacy in preventing a relapse of the disease. Study objectives were to assess the equivalence of the clinical activity index (CAI) under the two treatment modalities and to compare relapse rates, relapse-free times and global assessment. RESULTS: The start and end scores of the CAI demonstrated no significant difference (P = 0.12) between the two treatment groups. Relapse rates were 11.3% under mesalazine and 16.0% under E. coli Nissle 1917 (N.S.). Life table analysis showed a relapse-free time of 103 +/- 4 days for mesalazine and 106 +/- 5 days for E. coli Nissle 1917 (N.S.). Global assessment was similar for both groups. Tolerability to the treatment was excellent and did not differ. No serious adverse events were reported. CONCLUSIONS: From the results of this preliminary study, probiotic treatment appears to offer another option for maintenance therapy of ulcerative colitis. Additional support is provided for the hypothesis of a pathophysiological role for the intestinal environment in ulcerative colitis. PMID- 9354193 TI - Nicotine enemas for active ulcerative colitis--a pilot study. AB - BACKGROUND: Since transdermal nicotine is of value in the treatment of active ulcerative colitis but is often associated with side-effects, an alternative in the form of topical therapy with nicotine enemas has been developed. METHODS: In an open study, 22 patients with active colitis, all non-smokers, were asked to take a 100 mL enema containing 6 mg of nicotine every night for 4 weeks. Pre trial treatment using mesalazine (n = 16), oral prednisolone (8), cyclosporin (1) and azathioprine (1) was kept constant for the month prior to assessment and during the study period. Symptoms, with stool frequency, were recorded on a diary card and an endoscopy was performed with rectal biopsy at the beginning of the study and after 4 weeks. RESULTS: Seventeen of the 22 patients completed 1 month of treatment. Mean duration of relapse was 29 weeks, range 3-94. Sixteen of 17 improved their St Mark's score. Urgency and stool frequency improved in 12 patients, sigmoidoscopic and histological scores in 10. Three patients had a full remission of symptoms with normal sigmoidoscopy. Six of 10 with a partial response continued with the enemas for a second month and five showed further improvement with full remission in two. The enema appeared effective when added to conventional treatment and produced few side-effects. CONCLUSION: Topical nicotine therapy for ulcerative colitis may have a place in future management, but controlled studies are needed. PMID- 9354194 TI - A dose-ranging pharmacokinetic study of nicotine tartrate following single-dose delayed-release oral and intravenous administration. AB - BACKGROUND: Ulcerative colitis is predominantly a disease of non-smokers, and transdermal nicotine is therapeutic but often results in side-effects. Administration of nicotine tartrate as a liquid enema decreases systemic nicotine absorption and may be effective for treatment of active distal ulcerative colitis. Ileocolonic delivery of nicotine tartrate via a delayed release oral capsule would be the preferred route to deliver nicotine to the colon. AIM: To determine the bioavailability and pharmacokinetic parameters of delayed-release oral nicotine tartrate capsules (Eudragit S100 coated) at doses of 3 mg and 6 mg nicotine. METHODS: Twenty healthy human subjects received delayed-release oral nicotine tartrate at one of two doses (each group n = 10): 3 mg and 6 mg nicotine. All subjects also received intravenous nicotine tartrate (at a dose of 15 micrograms nicotine base/kg) during a separate study period. Serum nicotine concentrations were determined by gas chromatography-mass spectrometry. In addition, concentrations of serum cotinine (major nicotine metabolite) were determined by high-performance liquid chromatography in all samples for two subjects (both given 6 mg nicotine). Adverse reactions were determined by questionnaire. RESULTS: The mean bioavailabilities of nicotine after ileocolonic nicotine tartrate administration via delayed-release oral capsules at doses 3 mg and 6 mg nicotine were 41% and 42%, respectively. The ratios (after adjusting for nicotine dose) of cotinine area under the curve (AUC) for delayed-release oral nicotine to cotinine AUC for intravenous nicotine were 1.5 and 1.6 for the two subjects undergoing cotinine pharmacokinetics, demonstrating significant first pass metabolism. Serum nicotine concentrations did not predict adverse reactions. CONCLUSIONS: Nicotine tartrate delivered to the ileocolon as a delayed-release oral capsule at doses of 3 mg and 6 mg nicotine considerably reduced systemic nicotine bioavailability. This reduction in bioavailability appears to be a result of first-pass hepatic metabolism rather than poor mucosal absorption of nicotine. The therapeutic potential of an ileocolonic delivery formulation of nicotine tartrate, which can potentially limit toxicity by local delivery of high doses of nicotine, should be investigated in patients with ulcerative colitis. PMID- 9354195 TI - Inhibition of intragastric acidity in healthy subjects dosed with ranitidine 75 mg: a comparative study with cimetidine and placebo. AB - BACKGROUND: Despite the widespread use of over-the-counter H2-receptor antagonists little is known about their duration of action on human gastric acid secretion. There are studies reporting inhibitory effects for up to 9 h post-dose but few data beyond this period. METHODS: Profiles of 20-h intragastric acidity were measured simultaneously in 24 healthy subjects who were dosed (at 12.30 h) with either ranitidine 75 mg, cimetidine 200 mg or placebo in a three-way crossover study, according to a standard protocol. Five-millilitre aliquots of gastric juice were aspirated half-hourly during the day (0-10 h post-dose) and hourly overnight (10-20 h post-dose). pH was measured to three decimal places with a glass electrode. Weighted intragastric acidity (AUC/time) was calculated for both day- and night-times using 2.5-h intervals during the day and 5-h intervals at night. Statistical analysis was by ANOVA. RESULTS: The results are expressed as mean weighted intragastric acidity (mmol/L). (i) Daytime (0-10 h post-dose): when dosed with placebo the weighted intragastric acidity was 31.03, decreasing to 10.37 (P < 0.001 vs. placebo) and 16.23 (P < 0.001 vs. placebo) when treated with ranitidine and cimetidine, respectively. Ranitidine inhibited weighted intragastric acidity to a greater degree than cimetidine (P < 0.001) during this period. (ii) Night-time (10-20 h post-dose): when dosed with placebo the weighted intragastric acidity was 21.36 decreasing to 16.65 (P < 0.001 vs. placebo) when dosed with ranitidine and remaining unchanged at 20.03 (P = 0.886 vs. placebo) when dosed with cimetidine. Ranitidine inhibited weighted intragastric acidity to a greater degree than cimetidine (P = 0.010) during this period. A sub-analysis of the two 5-h intervals showed that compared to placebo, ranitidine inhibited weighted intragastric acidity significantly in the 10-15 h period. However, its effect in the 15-20 h period did not differ from placebo. CONCLUSIONS: In healthy subjects, the inhibitory effect of ranitidine 75 mg on intragastric acidity can be detected 10-15 h after an oral dose. By contrast, the inhibitory effect of cimetidine 200 mg seems to be restricted to the first 10 h. PMID- 9354196 TI - Nocturnal intragastric acidity after over-the-counter doses of famotidine, ranitidine or placebo. AB - AIM: To compare the inhibitory effects of over-the-counter doses of famotidine or ranitidine on nocturnal intragastric acidity in a placebo-controlled study. METHODS: Twelve-hour intragastric acidity was measured simultaneously in 24 healthy subjects who ate a standard meal at 18.30 h and were dosed (at 19.30 h) with either famotidine 10 mg, ranitidine 75 mg or placebo in a balanced three period crossover design. Five-millilitre aliquots of gastric juice were aspirated half-hourly 0-3 h post-dose, and then hourly until the end of the study. pH was measured with a glass electrode. Integrated pH (area under the curve (AUC) of the pH-time curve) was calculated for the intervals 0-12 h and 9-12 h post-dose. Statistical analysis was by ANOVA. RESULTS: In the 0-12 h post-dose period, when the 24 subjects were dosed with placebo, mean AUC was 2.12, increasing by 75% to 3.70 with famotidine (P < 0.001) and by 81% to 3.83 with ranitidine (P < 0.001). In the 9-12 h post-dose period, when the subjects were dosed with placebo, mean AUC was 2.13, increasing by 91% to 4.07 with famotidine (P < 0.001) and by 79% to 3.82 with ranitidine (P < 0.001). There was no significant difference between the pH-raising effects of famotidine and ranitidine in both periods. CONCLUSIONS: Famotidine and ranitidine in over-the-counter doses have a similar, sustained, effect on post-prandial nocturnal intragastric acidity in healthy subjects lasting up to 12 h after oral dosing. PMID- 9354197 TI - A prospective follow-up study of 5669 users of lansoprazole in daily practice. AB - BACKGROUND: Immediately after the introduction of the proton pump inhibitor lansoprazole, a 2-year follow-up study was started to evaluate patterns of use, safety and effectiveness of this drug in naturally occurring groups of patients in the Netherlands. Medical data were recorded by participating physicians while medication listing were provided by pharmacists. METHODS: The study was designed according to the Safety Assessment of Marketed Medicines guidelines. The only inclusion criterion was the use of lansoprazole prior to entry into the study. RESULTS: A total of 5669 lansoprazole users was included by 374 general practitioners and 117 specialists. Lansoprazole was mostly prescribed in patients with reflux oesophagitis (55.1%), 'gastritis' (26.8%) and duodenal ulcers (11.4%), sometimes as part of a Helicobacter pylori eradication therapy (8.5%). For their complaints most patients (91.1%) had previously used acid-related drugs. Improvement or disappearance of complaints was achieved in 88.9% and 90.5% of patients after 4 and 8 weeks of treatment, respectively. Diarrhoea (4.1%), headache (2.9%) and nausea (2.6%) were the most frequently reported adverse events. CONCLUSION: The patterns of use of lansoprazole in daily practice deviated from the recommendations in the information leaflet. Nevertheless, lansoprazole was found to be safe in this naturally occurring group of users. Effectiveness appeared to be comparable to results found in clinical trials of the registered indications for lansoprazole. PMID- 9354198 TI - Effect of omeprazole on the bioavailability of unmodified and phospholipid complexed aspirin in rats. AB - BACKGROUND: Treatment and prevention of non-steroidal anti-inflammatory drug induced gastropathy involve the concurrent use of antisecretory drugs. Recently, we have shown that the ability of these drugs to increase the intragastric pH to values > > pKa of NSAIDs compromises their therapeutic activity. In the present study, we evaluated the potential of omeprazole to interfere with the bioavailability of aspirin administered to rats either alone or complexed with the zwitterionic phospholipid, dipalmitoylphosphatidylcholine (DPPC). METHODS: Aspirin or aspirin/DPPC was administered intragastrically to rats pre-dosed with either saline or omeprazole. Concentrations of aspirin and salicylic acid in the blood and the gastric mucosa were assessed by HPLC and the 6-keto-PGF1 alpha gastric mucosal concentration by radioimmunoassay. RESULTS: Gastric absorption of aspirin and its relative bioavailability were reduced by an antisecretory dose of omeprazole; its inhibitory effect on gastric prostaglandin synthesis was consequently attenuated. However, these effects could be partly overcome if aspirin was administered as a complex with DPPC. CONCLUSIONS: These observations suggest that: (i) DPPC increases the lipid solubility and gastric permeability of NSAIDs; and (ii) neutralization of the gastric pH results in a shift of aspirin absorption toward the intestine where it could be degraded to salicylic acid. PMID- 9354199 TI - The influence of age, gender, Helicobacter pylori and smoking on gastric mucosal adaptation to non-steroidal anti-inflammatory drugs. AB - INTRODUCTION: Oral NSAIDs cause acute gastric injury that resolves, despite continued administration, by a process known as adaptation. Little is known about the factors that influence this process. METHODS: Sixty-two healthy volunteers were given a 28-day course of either etodolac 300 mg b.d. (13 subjects), naproxen 500 mg b.d. (23), enteric-coated diclofenac (10) or effervescent diclofenac 50 mg b.d. (16). All subjects were gastroscoped before and on days 1, 7 and 28 during drug administration, to assess gastric mucosal damage using a modified Lanza scale. Subjects were then divided into three categories: those who adapted completely, those who adapted incompletely and those who showed no adaptation. The proportion of subjects in each group was compared with respect to age, gender, smoking, the presence of Helicobacter pylori, and the NSAID prescribed. RESULTS: Fifty-nine subjects (median age 25.0 years, range 18-70) developed initial gastric injury to NSAIDs of whom 42 adapted completely, 13 adapted incompletely and four showed no evidence of adaptation. The mean age of subjects was lower in those who adapted (26.8 +/- 9.8 years) than those who adapted incompletely (32.5 +/- 10.3 years) and those who did not adapt (42.0 +/- 15.7 years, P = 0.01). There was no evidence of gender influencing adaptation. Of 17 H. pylori-positive subjects, a higher proportion had incomplete adaptation, with only nine subjects adapting completely (53% vs. 81%, P = 0.04). Sixteen subjects were smokers, of whom a greater proportion showed no evidence of adaptation (19% vs. 2%, P = 0.03). A smaller proportion of those who took naproxen (48%) adapted completely than those who took enteric-coated diclofenac (89%), effervescent diclofenac (75%) or etodolac (91%, P = 0.03). CONCLUSION: Some adaptation occurred in over 90% of subjects after 4 weeks dosing with an NSAID, but adaptation was less frequent in older subjects and in smokers. Complete adaptation occurred less frequently in H. pylori-positive subjects and in those who were given naproxen. PMID- 9354200 TI - A United States multicentre trial of dual and proton pump inhibitor-based triple therapies for Helicobacter pylori. AB - BACKGROUND: One-week proton pump inhibitor-based triple therapies are very popular in the US despite limited US data documenting efficacy. We assessed 1 week proton pump inhibitor triple therapies for Helicobacter pylori, and compared them to dual antibiotic therapies (to assess benefit of omeprazole) and to omeprazole-amoxycillin (to assess benefit of clarithromycin) in a large, randomized, US multicentre study. METHODS: Healthy subjects who were H. pylori positive by rapid serological test and 13C-urea breath test were randomly assigned to (i) omeprazole (O) 20 mg b.d. + amoxycillin (A) 1 g t.d.s. for 14 days (OA); (ii) A 1 g b.d. + clarithromycin (C) 500 mg b.d. for 7 days (AC); (iii) C 250 mg b.d. + metronidazole (M) 500 mg b.d. for 7 days (CM); (iv) O 20 mg b.d. + C 250 mg b.d. + M 500 mg b.d. for 7 days (MOC); or (v) O 20 mg b.d. + C 500 mg b.d. + A 1 g b.d. for 7 days (OAC). Repeat breath tests were done at 6 weeks to assess H. pylori status. RESULTS: Three hundred and two H. pylori positive subjects at 25 centres received medication. Intention-to-treat cure rate was significantly higher for OAC (82%) than for MOC (67%), CM (59%), AC (18%) or OA (58%), Per-protocol cure rates were 85% for OAC and 75% for MOC. Discontinuation of therapy due to a side-effect occurred in 0-3% of each study group. CONCLUSIONS: One-week twice-daily triple therapy with omeprazole, amoxycillin and clarithromycin provides the best rate of eradication of the five regimens studied. However, treatment in the US for 7 days may be unable to achieve eradication rates of > or = 90% with proton pump inhibitor-based triple therapy. PMID- 9354201 TI - Two omeprazole-based Helicobacter pylori eradication regimens for the treatment of duodenal ulcer disease in general practice. AB - BACKGROUND: Helicobacter pylori is the main acquired factor in the pathogenesis of duodenal ulcer disease. METHODS: This multicentre study conducted in 32 general practice centres in the UK and Ireland was a double-blind, placebo controlled, randomized, parallel-group comparison of triple therapy (n = 98: omeprazole 40 mg once daily and amoxycillin 1 g b.d. for 2 weeks, and metronidazole 400 mg t.d.s. for the first week) and dual therapy (n = 85: omeprazole 40 mg once daily and amoxycillin 1 g b.d. for 2 weeks, with placebo during the first week) for the eradication of H. pylori in patients with symptomatic duodenal ulcer disease. Patients who were successfully treated entered a follow-up phase for 12 months to assess symptomatic relapse and use of health-care resources. RESULTS: Eradication of H. pylori based on a second 13C urea breath test was successful in 95% (95% confidence interval (CI) = 90-100%) of patients receiving omeprazole triple therapy and 53% (95% CI = 41-65%) of those receiving omeprazole dual therapy (P < 0.0001 between groups, all data available analysis). The all-patients-treated analysis gave eradication rates of 80 and 44% for omeprazole triple therapy and omeprazole dual therapy, respectively. Symptomatic relapse occurred in 16% (18/116) of the H. pylori negative patients who entered the 12-month follow-up period, and there were significant reductions in the number of consultations, investigations and prescriptions relating to upper gastrointestinal symptoms compared with the 12 months prior to the eradication therapies (all P < 0.0001). The two treatment strategies were comparable in terms of the number of adverse events reported. CONCLUSIONS: Omeprazole triple therapy provides a highly effective treatment for the eradication of H. pylori infection in patients in general practice, with an adverse event profile similar to that seen with omeprazole dual therapy. The successful eradication of H. pylori with these omeprazole regimens results in a significant decrease in the use of health-care resources in the 12 months following treatment. PMID- 9354202 TI - The management of failed dual or triple therapy for Helicobacter pylori eradication. AB - BACKGROUND: After each treatment for Helicobacter pylori infection there is an eradication failure rate ranging from 5 to 50%. Thus, the best therapy schedule and treatment regimen sequence have still to be identified. METHODS: Patients with H. pylori infection were randomized to receive either a 1-week triple therapy of omeprazole 20 mg b.d., clarithromycin 250 mg b.d. and tetracycline 500 mg b.d. (OCT; 78 patients) or a 2-week dual therapy of omeprazole 20 mg b.d. and amoxycillin 1 g b.d. (OA; 75 patients). H. pylori infection at entry and eradication 4-6 weeks after therapy had ended were assessed by rapid urease test and histology on biopsies from the antrum and the corpus. When eradication did not occur with either the OCT or OA regimens, patients were switched over to the OA or OCT therapy, respectively. Eradication in these patients was assessed 4-6 weeks after conclusion of treatment by a further endoscopy. RESULTS: H. pylori eradication was achieved in 67.9% (95% CI = 57.6-78.3%) of patients treated with the OCT regimen and in 75.7% (95% CI = 65.9-85.5%) of patients treated with the OA therapy (chi 2 = 1.11; P = 0.29). Moreover, H. pylori eradication was achieved in 39.1% (95% CI = 19.2-59.1%) of patients re-treated with the OA regimen and in 88.9% (95% CI = 74.4-100%) of patients re-treated with the OCT therapy (chi 2 = 8.52; P = 0.003). Thus, the overall success rate 'per protocol' analysis in our study was 81.6% (95% CI = 72.9-90.3%) for the triple and dual therapy sequence and 97.3% (95% CI = 93.6-100%) for dual followed by triple therapy (chi 2 = 8.14; P = 0.004). CONCLUSIONS: Our data found that H. pylori eradication with OA therapy after OCT therapy failure was poor, while that obtained with OCT after OA therapy was good. PMID- 9354203 TI - Twice a day quadruple therapy (bismuth subsalicylate, tetracycline, metronidazole plus lansoprazole) for treatment of Helicobacter pylori infection. AB - BACKGROUND: Quadruple therapy (bismuth, metronidazole and tetracycline (BMT) + proton pump inhibitor) is touted as being > 95% effective, regardless of metronidazole resistance. We tested a 10-day b.d. quadruple therapy for treatment of H. pylori infection. METHODS: Anti-H. pylori therapy consisted of lansoprazole 15 mg b.d. plus tetracycline 500 mg b.d., metronidazole 500 mg b.d., and swallowable Pepto-Bismol caplets (2 b.d.) for 10 days. H. pylori status was evaluated by culture and histology before and 4 or more weeks after therapy. RESULTS: The cure rate for intention-to-treat was 70%. Treatment success was calculated overall and separately in relation to antimicrobial resistance patterns. The cure rate among the metronidazole-sensitive isolates was 89.7% (26 of 29) vs. 41.2% (7 of 17) of the metronidazole-resistant isolates (P < 0.005). Moderate (n = 1) or severe (n = 3) side-effects were experienced in four patients with only one withdrawing because of side-effects. CONCLUSION: Twice a day quadruple therapy is effective for metronidazole-sensitive strains but its usefulness is markedly reduced by the presence of pre-treatment metronidazole resistance. Twice a day quadruple therapy can be recommended in locations where background metronidazole resistance is uncommon. Possibly, 14-day therapy or a higher dosage of metronidazole provide better results with metronidazole resistant H. pylori. PMID- 9354204 TI - Clarithromycin, amoxycillin and H2-receptor antagonist therapy for Helicobacter pylori peptic ulcer disease in Korea. AB - BACKGROUND: Effective anti-Helicobacter pylori therapies with few side-effects are needed. We previously showed that the regimen of amoxycillin, clarithromycin and an H2-receptor antagonist was effective in the United States. The current study tested whether this therapy would also be successful in Korea. METHODS: Patients with gastric or duodenal ulcers received amoxycillin (750 mg t.d.s.) plus clarithromycin (500 mg t.d.s.) for 2 weeks and nizatidine 300 mg at bedtime for 6 weeks. Endoscopic examinations were performed before treatment and 4 or more weeks after ending antimicrobial therapy. H. pylori status was confirmed by rapid urease testing and histological examination of gastric antrum and corpus biopsies using the Genta stain. Antibiotic resistance was tested using the E-test method. Cure was defined as no evidence of H. pylori infection 4 or more weeks after ending therapy. RESULTS: Seventy-two patients (59 males and 13 females; mean age 46 years), including 35 with duodenal ulcers, 30 with gastric ulcers and seven with both, were studied. H. pylori infection was cured in 95.8% (69/72 patients; 95% CI = 88.3-99.1%). Two of the treatment failures had culture data and one had pre-treatment resistance to clarithromycin. Smoking did not have an adverse effect on therapy. Ten patients (15%) developed side-effects during treatment, but all were mild and did not require treatment interruption. No case of reinfection was noted during follow-up. CONCLUSION: The combination of amoxycillin, clarithromycin and an H2-receptor antagonist is effective in Korean patients with H. pylori infection. PMID- 9354205 TI - Short-course therapy with amoxycillin-clarithromycin triple therapy for 10 days (ACT-10) eradicates Helicobacter pylori and heals duodenal ulcer. ACT-10 Study Group. AB - BACKGROUND: Whilst the role of Helicobacter pylori eradication in managing duodenal ulcers has been established, consensus regarding the ideal regimen has not been achieved. METHODS: Patients with H. pylori-positive active duodenal ulcer were randomly assigned to receive triple therapy with amoxycillin 1000 mg b.d. + clarithromycin 500 mg b.d. + omeprazole 20 mg daily for 10 days (ACT-10) or dual therapy with clarithromycin 500 mg t.d.s. + omeprazole 40 mg daily for 14 days (Dual). No additional acid suppression was provided following eradication therapy. Endoscopy, with biopsy for culture and histology, as well as 13C-urea breath testing (13C-UBT) were performed pre-treatment to assess H. pylori infection. H. pylori eradication was established at 4-6 weeks follow-up with culture (2 antral, 1 corpus biopsies), histology (2 antral biopsies), and 13C UBT. Ulcer healing by endoscopy and change in clinical symptoms were also assessed at 4-6 weeks. RESULTS: Two hundred and sixty-seven (267) patients were randomized to ACT-10 (n = 137) or Dual therapy (n = 130). By per-protocol and intention-to-treat analyses, H. pylori eradication at 4-6 weeks follow-up was 91% (115/127) and 88% (120/136), respectively, for ACT-10 patients and 59% (68/115) and 55% (72/130), respectively, for Dual therapy patients (P < 0.001 for both analyses). Ulcer healing was high in both treatment groups: ACT-10, 93% (118/127) and 90% (122/136), respectively; and Dual therapy, 91% (104/114) and 85% (111/130), respectively. Pre-treatment resistance to clarithromycin was low (4%, 8/214) as compared to metronidazole resistance which was over 40%. Emergence of resistance to clarithromycin was observed in 2% of patients receiving ACT-10 and in 25% of those receiving Dual therapy. ACT-10 and Dual therapy patients experienced similar rates of drug-related adverse events (33% vs. 32%, respectively) and discontinuation from therapy due to an adverse event (1.5% vs. 5%, respectively). More than 90% of patients were compliant with each prescribed medication. CONCLUSION: In patients with active duodenal ulcer, a 10-day course of amoxycillin-clarithromycin-based triple therapy without additional acid suppression is highly effective in eradicating H. pylori and healing duodenal ulcer. PMID- 9354206 TI - Topical benzocaine anaesthesia lacks analgesic effects in painful non-acid oesophagitis. AB - BACKGROUND: No established treatment exists for pain relief in symptomatic non acid oesophagitis. One of the most common topical anaesthetics is benzocaine which has been demonstrated to produce excellent analgesia on oral mucous membranes. METHODS: In a prospective, placebo-controlled, balanced, single blinded study, 26 patients with retrosternal discomfort or odynophagia due to painful non-acid oesophagitis were treated either with oral benzocaine 0.75% solution (n = 14) or with benzocaine-free solvent (placebo, n = 12) at a daily dose of 20-40 mL, for up to 6 consecutive days (median 5.5 days). During the study period patients recorded subjective pain scores for both complaints on visual analogue scales. RESULTS: Benzocaine did not affect pain scores for any of the two symptoms, nor did it alter global subjective or objective assessment of therapy outcome in treated compared to untreated subjects (P > 0.05). There was a non-significant tendency for placebo patients to stop prematurely their study medication more often, because of lack of analgesic efficacy (P = 0.098). CONCLUSIONS: Topical benzocaine cannot be recommended for routine symptomatic pain relief in non-acid oesophagitis. By indirect evidence, it is assumed that pain perception of non-acid oesophagitis is not preferentially mediated by superficially located mucosal nociceptors. PMID- 9354207 TI - Treatment of reflux oesophagitis of moderate and severe grade with ranitidine or pantoprazole--comparison of 24-hour intragastric and oesophageal pH. AB - BACKGROUND: Proton pump inhibiting drugs strongly decrease gastric acid secretion and have proven more effective in the treatment of reflux oesophagitis than H2 receptor antagonists. METHODS: In a double-blind randomized trial, 24 patients with oesophagitis grade II (n = 15) and III (n = 9) were treated for 4 weeks with either ranitidine 150 mg b.d. (n = 13) or pantoprazole 40 mg o.m. (n = 11). Before the trial and on the last day of medication, 24-h intragastric pH and oesophageal pH profiles were performed. Healing was assessed by endoscopy. RESULTS: Pantoprazole increased median gastric pH from 1.7 to 3.9. Virtually no change in gastric pH was seen in the ranitidine group. Pantoprazole reduced the fraction time of pH < 4 in the oesophagus from 21% to 3% (P = 0.0005), and the median number of refluxes from 206 to 56 (P = 0.022). Oesophageal acid exposure was not decreased by ranitidine. Healing of the oesophagitis was seen in 6/11 cases after pantoprazole and in 3/13 cases after ranitidine (N.S.) CONCLUSION: In patients with oesophagitis of moderate and severe grade, pantoprazole 40 mg o.m. decreases intragastric acidity and gastro-oesophageal acid reflux more effectively than ranitidine 150 mg b.d. PMID- 9354208 TI - Effects of 3-days' intake of a sustained-release preparation of the nitric oxide donor, isosorbide dinitrate, on oesophageal motility. AB - BACKGROUND: Nitric oxide plays an important role in gastrointestinal motility. We evaluated the effects of a sustained-release preparation of the nitric oxide donor isosorbide dinitrate on swallow-initiated oesophageal contractions and the lower oesophageal sphincter. METHODS: Twelve healthy men, aged 23-32 years, received, at 1-week intervals and under random double-blind conditions, for 3 days either 20 mg isosorbide dinitrate, 40 mg isosorbide dinitrate or placebo twice daily (b.d.). One hour after a further dose on day 4, oesophageal motility was recorded for 30 min using a multilumen catheter with a Dent sleeve straddling the lower oesophageal sphincter and side-hole openings 0, 3, 6 and 9 cm proximal to the sleeve. Contractile responses to twelve 5-mL water swallows were evaluated. RESULTS: Amplitude, duration, propagation velocity and onset latency of oesophageal contractions were not affected by either dosage of isosorbide dinitrate. Lower oesophageal sphincter resting pressure was significantly lower after 40 mg (15.1 mmHg +/- 1.2 S.E.M.) and 20 mg isosorbide dinitrate b.d. (15.0 +/- 1.0 mmHg) than after placebo (17.9 +/- 1.7 mmHg; P < 0.025). Headache was reported by all subjects on 40 mg isosorbide dinitrate, seven subjects on 20 mg and by one on placebo. CONCLUSIONS: Twenty and 40 mg sustained-release isosorbide dinitrate twice daily had no effect on swallow-initiated oesophageal contractions but decreased lower oesophageal sphincter resting pressure. PMID- 9354209 TI - Effects of oral rabeprazole on oesophageal and gastric pH in patients with gastro oesophageal reflux disease. AB - BACKGROUND: This study examined the dose-response effects of the new proton-pump inhibitor rabeprazole on oesophageal and gastric pH in patients with gastro oesophageal reflux disease. METHODS: This study had a single-centre, double blind, randomized, two-way crossover design. Twenty patients were treated for two 7-day periods separated by a 7-10-day washout period. Patients were randomly assigned to receive either 20 mg of rabeprazole once daily during the first treatment period and 40 mg once daily during the second treatment period, or 40 mg during the first treatment period and 20 mg during the second treatment period. The primary efficacy variable was oesophageal acid exposure determined by 24-hour ambulatory pH monitoring. Acid-reflux time was defined as the percentage of time over 24 h that oesophageal pH was < 4. A dosage was considered effective if reflux time was reduced to < 6%, a number which has been our internal laboratory reference. RESULTS: Both rabeprazole 20 mg and 40 mg, given once daily, normalized reflux time, with decreases of 79% and 92% in acid exposure by day 7. Both dosages also decreased the mean total number of reflux episodes and the number of episodes lasting > 5 min, with no significant differences between dosages for any reflux parameter. Mean gastric pH increased with 20 mg from 1.86 at baseline to 3.71 on day 1 and 4.17 on day 7. Rabeprazole 40 mg once daily increased gastric pH from 2.01 to 4.37 on day 1, and to 4.65 on day 7. Safety analyses revealed no significant acute side-effects for either dosage. CONCLUSIONS: Pathological oesophageal acid exposure was normalized with both 20 mg and 40 mg dosages of rabeprazole, and the effects of these two doses did not differ. Rabeprazole was well-tolerated in this short-term study. PMID- 9354210 TI - Lack of dose-response with Pancrease MT for the treatment of exocrine pancreatic insufficiency in adults. AB - BACKGROUND: Choosing the optimum pancreatic enzyme replacement therapy for patients with exocrine insufficiency remains a problem. An enteric coated enzyme microsphere pancreatic enzyme preparation (Pancrease) has been marketed with several levels of lipase activity, implying that there is a dose-response relationship between dose and effectiveness such that the high potency form appears to be the most cost effective. METHODS: In a randomized, single-blind, cross-over study, we evaluated the effectiveness of a commercial enzyme preparation with different amounts of lipase per dosage unit in adults with exocrine pancreatic insufficiency. Patients received a diet comprising 100 g fat each day for 6 days. With each meal (three per day) they received two capsules of either Pancrease MT4 (8000 unit lipase), Pancrease MT10 (20,000 units lipase), Pancrease MT16 (32,000 units lipase) or placebo. A 72-h quantitative faecal collection was carried out for the last 3 days of the 6-day period. RESULTS: There was a reduction in faecal fat excretion with each of the preparations compared to placebo. The difference failed to reach significance with the 8000 units lipase preparation (P > 0.05) but was significant (P = 0.02) with the 20,000 units lipase and the 32,000 units lipase preparations (faecal fat excretion: placebo = 42.1 +/- 29 g, lipase 8000 = 22.1 +/- 7.3 g, lipase 20,000 = 10.2 +/- 4.5 g and lipase 32,000 = 15.8 +/- 12.5 g, P < for 20,000 units and 32,000 units lipase compared to placebo). CONCLUSION: A dose-response relationship between the amount of lipase administered with each meal and a reduction in faecal fat was not evident. The most potent preparation did not provide additional benefits compared to the less expensive lower potency dosage form. PMID- 9354211 TI - Tilidine does not affect human sphincter of Oddi motility--a randomized, controlled study. AB - AIM: To investigate the effects of intravenous pentazocine and tilidine on sphincter of Oddi motility. METHODS: Twenty patients with suspected sphincter of Oddi dysfunction were enrolled in a prospective, double-blind study. Sphincter of Oddi motility was assessed by means of endoscopic manometry after injection of 0.9% saline, as well as after randomized dosing with either 30 mg pentazocine i.v. (n = 10) or 50 mg tilidine i.v. (n = 10). RESULTS: Pentazocine significantly increased the sphincter of Oddi baseline pressure from 32 +/- 21 mmHg (saline) to 41 +/- 19 mmHg (P = 0.002), whereas tilidine did not alter the sphincter baseline pressure (34 +/- 15 mmHg saline vs. 36 +/- 16 mmHg tilidine, P = 0.16). Furthermore, pentazocine increased the phasic sphincter contraction amplitude (108 +/- 16 mmHg saline vs. 121 +/- 18 mmHg pentazocine, P = 0.004), but tilidine was without any effect (125 +/- 24 mmHg saline vs. 125 +/- 21 mmHg tilidine, P = 0.93). The phasic sphincter of Oddi contraction frequency and duration were not influenced either by pentazocine or by tilidine. CONCLUSION: In contrast to 30 mg of pentazocine, 50 mg of tilidine does not affect sphincter of Oddi motility. Therefore, tilidine can be used during endoscopic manometry and for analgesia in pancreatobiliary disease. PMID- 9354212 TI - Haemodynamic responses to a combination of terlipressin and octreotide in portal hypertensive rats. AB - BACKGROUND: An enhancement of the haemodynamic effects of terlipressin by octreotide (and vice versa) may be useful in the treatment of portal hypertension. The aim of this study was to investigate the short-term effects of terlipressin, octreotide or a combination of these substances on splanchnic and systemic haemodynamics in rats with portal vein stenosis. METHODS: Eight rats received an intravenous (i.v.) infusion of isotonic saline (10 microL/min for 15 min). Eight rats received terlipressin first (0.05 mg/kg) and then an i.v. infusion of octreotide (8 micrograms.h/kg for 15 min) 15 min later. Eight other rats first received an i.v. infusion of octreotide and then terlipressin 15 min later. Splanchnic and systemic haemodynamics (radioactive microsphere method) were measured after saline, after terlipressin or octreotide alone, and after the combined treatments. RESULTS: Terlipressin and octreotide alone significantly decreased portal pressure, portal tributary blood flow and cardiac index. Terlipressin, but not octreotide, significantly increased heptocollateral vascular resistance and arterial pressure. Octreotide administration in rats pre treated with terlipressin did not change portal pressure, caused portal tributary blood flow to increase and decreased hepatocollateral vascular resistance; it also decreased arterial pressure but not cardiac index. Terlipressin administration in rats pre-treated with octreotide further decreased portal pressure, portal tributary blood flow and increased hepatocollateral vascular resistance; terlipressin increased arterial pressure and further decreased cardiac index. CONCLUSIONS: In rats with portal vein stenosis, octreotide decreased short-term splanchnic and systemic vasoconstriction due to terlipressin. In contrast, terlipressin enhanced the splanchnic and systemic vasoconstriction due to octreotide. Thus, the haemodynamic responses to the combination of octreotide and terlipressin depend on the order of administration of these substances. PMID- 9354213 TI - Oral or rectal administration of drugs in IBD? PMID- 9354214 TI - Helicobacter pylori and gastric acid secretion. PMID- 9354215 TI - Helicobacter pylori eradication. PMID- 9354217 TI - Reducing patient delay in seeking treatment. PMID- 9354216 TI - H. pylori eradication by LAC. PMID- 9354218 TI - Aortic valve replacement with a pulmonary autograft: case studies of the Ross procedure. AB - The Ross procedure is an old surgical technique that is gaining popularity some 30 years after its introduction. The patient's own pulmonary valve is essentially used as a "spare part" to replace the diseased aortic valve. The Ross procedure has become an accepted and attractive option for patients with a life expectancy of more than 20 years and for patients in whom long-term treatment with anticoagulants is undesirable or problematic. Careful preoperative evaluation and postoperative management, as well as proficient surgical techniques, are all necessary to achieve good outcomes. PMID- 9354219 TI - Lung volume reduction: surgical treatment for emphysema. AB - BACKGROUND: Lung volume reduction surgery is currently being investigated as a method of improving the respiratory function of patients with end-stage emphysema. PURPOSE: This article reviews the current literature on lung volume reduction surgery and proposes a multidisciplinary team approach to postoperative management. METHODS: We did a MEDLINE search and retrieved relevant articles. We selected and reviewed nine medical articles published in 1993, 1995, and 1996; one medical article from 1959; and one of four nursing articles. Overall, the articles describe the different techniques of median sternotomy and video assisted thoracoscopic surgery and unilateral versus bilateral procedures. CONCLUSION: Lung volume reduction surgery is beneficial, but further investigation is required. PMID- 9354220 TI - Analysis of risk factors for omental herniation associated with removal of peritoneal catheters. AB - BACKGROUND: Peritoneal catheters may be used routinely in children undergoing cardiac surgery. Removal of the catheter is often complicated by omental herniation, which can cause intraperitoneal bleeding, peritonitis, and bowel obstruction. Instillation of saline into the catheter before removal is used in some institutions as a preventive measure, but the practice has never been investigated. OBJECTIVE: To determine the effectiveness of instilling saline into peritoneal catheters before their removal in reducing the occurrence of omental herniation and to determine risk factors for omental herniation. METHODS: A total of 404 patients with peritoneal catheters in place were randomized to either the control or the study group. The study group (n = 204) had saline (1 mL/kg; maximum, 10 mL) instilled into the catheter before the catheter was removed; the control group (n = 200) did not. Extrusion or no extrusion of omentum was recorded. Other data collected included the child's weight, the length of time the catheter was in place, whether peritoneal dialysis was performed, and whether the child was pharmacologically paralyzed when the catheter was removed. RESULTS: Extrusion of omentum occurred in 39% of the study group and in 33% of the controls. Results of logistic regression analysis suggested that omental herniation was more common in smaller children, children whose catheters remained in place longer, and children who were not pharmacologically paralyzed when the catheter was removed. CONCLUSIONS: Instillation of saline does not appear to reduce the occurrence of omental herniation. Further research into strategies to reduce this complication is recommended. PMID- 9354221 TI - Nausea and vomiting in patients undergoing radiofrequency catheter ablation. AB - BACKGROUND: Radiofrequency catheter ablation is a technique for curing cardiac arrhythmias by destroying tissue that is a critical part of the targeted electrical circuit. Nausea and vomiting have been observed after use of this technique, but no study to date has examined these complications. OBJECTIVES: To determine the incidence of nausea and vomiting after radiofrequency catheter ablation, determine factors related to nausea and vomiting, and evaluate the antiemetic efficacy of promethazine given during the procedure. METHODS: Medical records for 369 cases of radiofrequency catheter ablation performed in a 4-year period at a university medical center were reviewed. RESULTS: Nausea after the procedure was documented in 22% of cases. Logistic regression analysis based on variables significant in bivariate analysis revealed that younger age, female sex, and longer procedure duration were significantly and independently related to nausea. Vomiting after the procedure occurred in 13% of cases. Logistic regression revealed that younger age and longer procedure duration were significantly and independently related to vomiting. Further analysis showed that patients who received larger doses of promethazine relative to the dose of fentanyl had a significantly lower incidence of vomiting. CONCLUSIONS: Nausea and vomiting occurred in a considerable number of cases. Female sex, younger age, and longer procedure duration increased the risk of nausea, whereas only age and procedure duration were associated with vomiting. This study may guide clinicians to use a prophylactic antiemetic in patients undergoing radiofrequency catheter ablation, especially patients at increased risk for nausea and vomiting. PMID- 9354222 TI - Implantable cardioverter defibrillators: physical and psychosocial outcomes. AB - BACKGROUND: The long-term outcomes of living with an implantable cardioverter defibrillator are an important consideration in recovery. However, little is known about physical and psychosocial outcomes beyond 1 year after implantation. OBJECTIVE: To describe the long-term physical and psychosocial adaptation of persons who have had an implantable cardioverter defibrillator for approximately 2 years or more. METHODS: This nonexperimental cross-sectional study used telephone interviews to ascertain the responses of 80 recipients of implantable cardioverter defibrillators to physical and psychosocial questionnaires to explore the long-term outcomes of living with the devices. Subjects eligible for inclusion were selected from the files of an arrhythmia clinic. RESULTS: Hierarchical regression analysis showed that subjects who are not emotional are likely to be more physically active, especially if they are young and male, and that subjects who tend to be emotional are likely to be psychologically distressed and have poorer social and domestic adaptation. Furthermore, use of emotions was a positive predictor of psychological distress and poor social and domestic adaptation. Subjects reported the use of both emotion- and problem focused coping. Subjects' scores on physical and psychosocial functioning were comparable to scores reported in the literature for patients who have had myocardial infarction or dysrhythmia. CONCLUSIONS: Emotional responses to distress were predictive of little physical activity and psychological distress. Furthermore, young recipients of implantable cardioverter defibrillators and men were predicted to be physically active. Persons who have had an implantable cardioverter defibrillator for approximately 2 years or more can anticipate that their physical and psychosocial functioning will be similar to that of patients who have myocardial infarction or dysrhythmia. PMID- 9354223 TI - Hemodynamic monitoring: a comparison of research and practice. AB - BACKGROUND: Measurement of hemodynamic parameters is a common practice and is well researched, but little information is available on the translation of the research into actual practice. OBJECTIVES: To determine (1) the current practice of hemodynamic measurement in relation to positioning of patients and (2) the barriers to use of research related to hemodynamic monitoring. METHODS: A stratified, random, national sample of 1000 members of the American Association of Critical-Care Nurses was surveyed by mail about hemodynamic monitoring procedures related to the positioning of patients and beds, technique used to determine the position of the bed, number of measurements of cardiac output, use of iced versus room-temperature injectate, use of printed or digital information, nurses' input into written procedures, and barriers to research utilization. RESULTS: In actual practice, 24.1% of the respondents always keep the bed flat when measuring pulmonary artery pressures, 55.0% elevate the head of the bed 30 degrees or less, 80.7% always have patients supine for measurements, and 13.3% place patients in lateral or other positions. Unit policy dictated the flat position in 25.8% of the sample; nurse managers (41.4%), staff nurses (27.5%), and staff committees (31.2%) were involved in writing the procedure. Respondents viewed research related to hemodynamic monitoring procedures as relevant and valuable. The greatest barriers to utilization were lack of staff support for implementation, insufficient time to implement research findings, and feelings among nurses that they lacked authority to implement change. CONCLUSIONS: Research findings are generally being implemented at the bedside, although not completely or consistently. Minimizing the barriers to use of research is within the scope of nurses' practice. PMID- 9354224 TI - Placement of cardiac electrodes: written, simulated, and actual accuracy. AB - BACKGROUND: Research has shown that critical care nurses show low accuracy on written tests of placement of electrodes, yet it is unknown how this low accuracy translates into placement of electrodes on actual patients. OBJECTIVES: To determine if accuracy scores differ between three methods (written knowledge, simulated clinical practice, actual clinical practice) of evaluating placement of continuous ECG electrodes by a group of cardiac care nurses. METHODS: A standardized scoring diagram was used with three different methods of measuring the accuracy of 44 nurses who worked on a telemetry unit or medical ICU in placing continuous ECG electrodes. The three methods were (1) written knowledge- placement of stickers on a diagram of a torso, (2) simulated clinical practice- placement of electrodes on a human model, and (3) actual clinical practice- placement of electrodes on an assigned patient. RESULTS: For the total diagram score (maximum score = 11), no significant differences among groups were found. For the V1 subscale score (maximum score = 4), a significant difference among groups was found: Scheffe's test showed that the significant difference was between simulated and actual clinical practice. Percentages of nurses achieving the maximum, or accurate, score were 18% for written knowledge, 25% for simulated clinical practice, and 9% for actual clinical practice. CONCLUSIONS: Although total scores did not differ among groups, the mean scores indicate that placement of electrodes was not accurate by any method. This finding has implications for how electrode placement is taught to nurses and for the accountability of nurses for placement of electrodes on their patients. PMID- 9354225 TI - Estimating ischemic burden: comparison of two formulas. AB - BACKGROUND: Complex formulas based on elevations of the ST segment on the ECG allow noninvasive estimation of ischemic burden. However, the formulas require elaborate computations that make them clinically useless, particularly in critical and emergent situations. Estimation of the amount of ischemic myocardium is useful in monitoring effects of therapeutic interventions. OBJECTIVE: To compare a simplified formula that sums ST-segment deviations in 12 ECG leads with formulas widely validated in clinical trials: anterior ischemic area = 3[1.5(number of leads ST increases) -0.4]; inferior ischemic area = 3[0.6(sigma ST increases II, III, aVF) +2.0]. METHODS: Data were collected from 46 patients undergoing angioplasty of the left anterior descending branch of the coronary artery or the right coronary artery who had changes in the ST segment denoting ischemia during balloon inflation. Absolute values of ST-segment elevations or depressions in the 12 standard ECG leads with a minimum deviation of 0.5 mm were added to determine the sum of the ST-segment deviations during ischemia of the left anterior descending branch of the coronary artery and ischemia of the right coronary artery. This sum was compared with the anterior and inferior ischemic area scores. The Pearson Product-Moment Correlation Coefficient was used to measure the association between the scores. RESULTS: The sum of ST-segment deviations for the left anterior descending branch correlated with the anterior ischemic area score, and the sum of ST-segment deviations for the right coronary artery correlated with the inferior ischemic area score. CONCLUSIONS: The complex, but validated, formulas for anterior ischemic area and inferior ischemic area correlate well with the simpler sum of ST-segment deviations. Because the simpler sum is more clinically useful, it may provide an alternative for monitoring ischemic burden. PMID- 9354226 TI - Management of incessant ventricular tachycardia in a 39-year-old woman with cardiomyopathy. PMID- 9354227 TI - Sudden cardiac death--preventable--reversible. AB - SCD is defined as unexpected death due to cardiac causes that occurs within 1 hour of acute symptoms. SCD can be reversed with the use of an ICD. These devices now can be implanted by catheter techniques, obviating thoracotomy. SCD is preventable. The incidence of SCD can be significantly reduced by addressing the fundamental pathophysiology of SCD, which primarily is CAD. Our combined and aggressive implementation of preventive regimens to reduce the risk of cardiac events will save lives. These measures include diet, weight reduction, smoking cessation, regular exercise, and therapeutic drugs. Amiodarone, although effective in preventing lethal ventricular arrhythmias, has not matched the long term results of the ICD in the successful management of SCD. PMID- 9354228 TI - Typical and atypical clinical signs and symptoms of myocardial infarction and delayed seeking of professional care among blacks. PMID- 9354229 TI - Is rhodamine 123 an appropriate fluorescent probe to assess P-glycoprotein mediated multidrug resistance in vinblastine-resistant CHO cells? AB - Cellular drug resistance, which involves several mechanisms such as P glycoprotein (P-gp) overexpression, kinetic and metabolic quiescence, or the increase in the intracellular levels of glutathione, limits the effectiveness of cancer treatment. It has been reported that functional assessment of the cationic dye rhodamin 123 (Rho123) efflux reveals accurately the drug-resistant phenotype. To study cellular drug resistance, we have obtained a CHO-K1 derived cell line resistant to vinblastine by means of multistep selection. This cell line (CHOVBR) displays high reactivity with a monoclonal antibody (MAb) (C219) directed against an internal domain of P-gp, and an active Rho123 efflux, as shown by parallel flow cytometric and fluorometric assays. However, under similar experimental conditions, the drug-sensitive parental cell line CHO-K1 (as well as the myeloblastic KG1 and KG1a cell lines), was also able to pump Rho123 out. These parental CHO-K1 cells had a very low reactivity against the C219 Mab, as confirmed by Western blot analysis. Both vinblastine and verapamil inhibited Rho123 efflux in CHO-K1 cells, but had no effect on CHOVBR cultures. Also, deprivation of vinblastine for one month did not affect Rho123 efflux in these cells. Our results suggest that the activity of P-gp appears to be essential, but not sufficient to confer drug resistance, and that Rho123-based functional assays of drug resistance should be evaluated for each cellular experimental model. PMID- 9354230 TI - Inhibition of the Ser-Thr phosphatases PP1 and PP2A by naturally occurring toxins. AB - The okadaic acid class of naturally occurring toxins is a structurally diverse group of molecules that inhibit the protein phosphatases PP1 and PP2A. Studies providing information about the mode of binding between the toxins and the phosphatases contribute to an overall understanding of the signal transduction pathways in which the phosphatases are involved. PMID- 9354231 TI - A molecular modeling analysis of the binding interactions between the okadaic acid class of natural product inhibitors and the Ser-Thr phosphatases, PP1 and PP2A. AB - We have proposed computer-generated models of the catalytic subunits of the serine-threonine protein phosphatases PP1 and PP2A complexed with their endogenous substrate phospho-DARPP-32, and several known naturally occurring inhibitors. This study is part of an overall effort to elucidate the signal transduction pathways in which PP1 and PP2A may play an important role. PMID- 9354232 TI - Synthesis and alpha-adrenoceptor blocking activity of the enantiomers of benzyl (2-chloroethyl)-[2-(2-methoxyphenoxy)-1-methylethyl]amine hydrochloride. AB - The enantiomers of benzyl-(2-chloroethyl)-[2-(2-methoxyphenoxy) -1 methylethyl]amine hydrochloride (1, CM18) were synthesized and studied pharmacologically for their irreversible antagonism at rat vas deferens alpha adrenoceptors. In addition, assignment of the absolute configuration of the two enantiomers of 1 was made by X-ray crystallographic analysis performed on the intermediate amine (+)-2 hydrochloride. The enantiomer (R)-(+)-1 [(R)-(+)-CM18] (a) had a 10-fold preferential blocking activity for alpha 1-versus alpha 2 adrenoceptors, (b) discriminated, like racemic 1, between two possible alpha 1 adrenoceptor subsites/subtypes, with a selectivity ratio of 6.5 and (c) was 10-23 times as potent as the (S)-(-)-enantiomer at alpha 2- and alpha 1-adrenoceptors. Thus, it may be a valuable tool for the characterization of rat vas deferens alpha 1-adrenoceptor subtypes. PMID- 9354233 TI - Structure-activity studies of the inhibition of serine beta-lactamases by phosphonate monoesters. AB - A new series of phosphonyl derivatives has been prepared and tested for inhibition of serine (class A and C) beta-lactamases. Variations of the leaving group in a series of methyl phosphonates showed that leaving groups better than the previously employed p-nitrophenoxide could give more effective inhibitors. Inclusion of a negative charge in the leaving group did not, per se, lead to better inhibitors. Aryl phosphonates appeared more effective than those with electronically comparable but smaller leaving groups. The combination of a good leaving group, 2,4-dinitrophenoxide, with an amido side-chain, phenylmethylsulfonamido--the latter rather than phenylacetamido in order to increase the stability of the compound with respect to intramolecular nucleophilic catalysis of hydrolysis by the amide group--did not yield overall a better inhibitor than previously employed p-nitrophenyl phosphonates. These results give the first indication of specific interactions between a beta lactamase and the leaving group of a phosphonate inhibitor. Only one enantiomer of a chiral thiophosphonate, presumably the Rp isomer, was an effective inhibitor. Addition of either a D- or a L-methyl group to the methylene group of a p-nitrophenyl amidomethylphosphonate did not enhance the inhibitory ability of the phosphonate. Class A beta-lactamases remain refractory to phosphonates. PMID- 9354234 TI - E-64 analogues as inhibitors of cathepsin B. On the role of the absolute configuration of the epoxysuccinyl group. AB - A series of trans-epoxysuccinyl-peptide derivatives based on the natural inhibitor E-64 were synthesized in the (2R,3R) and (2S,3S) configuration in order to analyze the role of the stereochemistry of this residue in dictating inhibitory potency and selectivity for cysteine proteases. We confirmed that binding of E-64 like trans-epoxysuccinyl compounds is remarkably favored by the (2S,3S) configuration, but we also found that CA030-type compounds are stronger inhibitors in the (2R,3R) configuration than the related diastereomers. Consequently, the structural requirements for exploiting both the S and S' subsites are not additive and a structure-based design of bis-peptidyl derivatives of trans-epoxysuccinic acid to increase selective inhibition becomes even more difficult. Additional contrasting effects were observed for the pH optima required in the electrostatic interactions at the S and S' subsites. PMID- 9354235 TI - Myrosinase-generated isothiocyanate from glucosinolates: isolation, characterization and in vitro antiproliferative studies. AB - Epidemiological and pharmacological studies have shown that colorectal cancer development could be reduced by consuming vegetables that contain glucosinolates. In view of this the effect of some glucosinolates and their isothiocyanate (ITC) derived products on in vitro cell growth was studied. We report the isolation and characterization of ITCs derived from glucosinolates by using HPLC, GC-MS, and NMR techniques. The in vitro activity of ITCs on human erythroleukemic K562 cells has been investigated by using two alternative approaches: the in situ and pre mix methods. No differences in antiproliferative activity were found comparing the effect of ITCs produced either of these methods. In the experimental conditions used, the production of ITCs from glucosinolates is almost quantitative as confirmed by HPLC or GC-MS analysis. The ITCs' inhibitory activity on K562 cells growth is particularly evident in the cases of ITCs derived from sinigrin, progoitrin, epi-progoitrin, glucotropaeolin and glucocheirolin. Finally, the antiproliferative activity of the ITCs obtained from glucoraphenin, taken as an example, was determined on other tumor cell lines with a different origin and hystotype. Considering the antiproliferative activity found for ITCs these compounds could be considered potentially responsible for the reduction of colorectal cancer associated with diets rich in cruciferous vegetables. Further studies will be aimed at the possible application of glucosinolate-derived products as chemopreventive cancer agents. PMID- 9354236 TI - Structure-activity relationship for DNA topoisomerase II-induced DNA cleavage by azatoxin analogues. AB - Eighteen analogues of the nonintercalative DNA topoisomerase II (topo II)-active epipodophyllotoxin-ellipticine hybrid, azatoxin, were synthesized and evaluated for their ability to induce topo II-mediated DNA strand breaks in vitro. In general, the SAR profile of the azatoxins showed more homology with that of the epipodophyllotoxins than with the ellipticines. Of the compounds studied, only fluoro substitution at the 8-, 9, and 10-positions of azatoxins enhanced activity, with 9-fluoroazatoxin being the most active compound in this series. PMID- 9354237 TI - 10-Formyl-5,8,10-trideazafolic acid (10-formyl-TDAF): a potent inhibitor of glycinamide ribonucleotide transformylase. AB - The synthesis of 10-formyl-5,8,10-trideazafolic acid (3) as a potential inhibitor of glycinamide ribonucleotide transformylase (GAR Tfase) is reported. The target compound was prepared by a convergent synthesis utilizing the alkylation of hydrazone 5 with benzylic bromide 6 to construct the core heterocycle 7. The aldehyde 3 and related agents were evaluated as inhibitors of purN GAR Tfase and avian AICAR Tfase. Compound 3 exhibited potent inhibition of GAR Tfase with a Ki of 0.26 +/- 0.05 microM. In contrast, 3 exhibited more moderate inhibition of aminoimidazole carboxamide ribonucleotide transformylase (AICAR Tfase), with Ki of 7.6 +/- 1.5 microM. PMID- 9354238 TI - Functionalized analogues of 5,8,10-trideazafolate as potential inhibitors of GAR Tfase or AICAR Tfase. PMID- 9354239 TI - Functionalized analogues of 5,8,10-trideazafolate: development of an enzyme assembled tight binding inhibitor of GAR Tfase and a potential irreversible inhibitor of AICAR Tfase. AB - A set of inhibitors 3 and 4 of GAR and AICAR Tfase based on the TDAF core which contain an sp2 C-10 carbon atom replacing N-10 of the natural cofactor are detailed. Both possess electrophilic olefins and the potential of trapping the reacting amine of the substrates GAR and AICAR by a Michael addition at the enzyme active site to provide an enzyme-assembled tight binding inhibitor. While these agents did not display such characteristics and served as simple competitive inhibitors of GAR Tfase and AICAR Tfase, inhibitor 15 prepared in the conversion of 3 to 4 may provide an enzyme-assembled tight binding inhibitor of GAR Tfase upon reaction with the substrate GAR and may inactivate AICAR Tfase by virtue of alkylation of an active site residue. PMID- 9354242 TI - Effect of stereochemistry on the transport of Aca-linked beta-turn peptidomimetics across a human intestinal cell line. AB - Transcellular transport is one of the most important barriers facing the development of new therapeutic agents. However, little is known about the specific effects of structure and particularly stereochemistry on cell permeability. An attractive in vitro model has been developed for the direct assessment of cell transport, using the immortalized human epithelial cell line, Caco-2. The present study assesses the effects of stereochemistry on transport in a commonly used beta-turn model system. Thus, L,L- and L,D-Ala-Ala were cyclized with aminocaproic acid, resulting in macrocycles in which the dipeptides correspond to the i + 1 and i + 2 positions of a beta-turn. The transport of these dipeptides across a Caco-2 cell monolayer was determined, along with corresponding acyclic models (L,L- and L,D-CH3CH2C(O)-Ala-Ala-n-Pr). The transport studies were carried out in the presence and absence of verapamil, a known inhibitor of the apically polarized efflux system present in Caco-2 cells. Both apical-->basolateral and basolateral-->apical transport were measured. Measurements made in the presence of verapamil showed that the cyclic peptides experienced a ca. 4-5-fold difference in intrinsic flux depending on stereochemistry, with the L,D isomer being transported at a higher rate. These differences disappeared in the acyclic cases examined (permeability coefficient ratios of the L,D/L,L isomers were 1.04-1.13). These observations are discussed in terms of the conformations and hydrogen-bonding characteristics of the compounds as determined by NMR spectroscopy. PMID- 9354240 TI - Abenzyl 10-formyl-trideazafolic acid (abenzyl 10-formyl-TDAF): an effective inhibitor of glycinamide ribonucleotide transformylase. AB - The synthesis of N-[7-(2-amino-3,4-dihydro-4-oxo-quinazolin-6-yl) -6-formyl-1-oxo heptyl]-L-glutamic acid (2, abenzyl 10-formyl-5,8,10-trideazafolic acid) as a potential enzyme-assembled tight binding inhibitor of glycinamide ribonucleotide transformylase (GAR Tfase) or aminoimidazole carboxamide ribonucleotide transformylase (AICAR Tfase) is reported. The inhibitor was prepared by a convergent synthesis utilizing the sequential alkylations of acetaldehyde dimethylhydrazone with 6 and 8. The agent exhibited effective inhibition of GAR Tfase (Ki = 4.5 +/- 0.3 microM) and more modest inhibition of AICAR Tfase (Ki = 42 +/- 11 microM). PMID- 9354241 TI - Multisubstrate analogue based on 5,8,10-trideazafolate. PMID- 9354243 TI - Libraries of opiate and anti-opiate peptidomimetics containing 2,3 methanoleucine. AB - A library of 96 peptides/peptidomimetics was prepared, in which half was based on the YGGFL-NH2 sequence, while the remainder were derivatives of a presumed anti opiate peptide, YGGFLRF-NH2. Of the 48 compounds in each half of the library, 32 contained a stereoisomer of 2,3-methanoleucine substituted for Leu5. Binding of the YGGFL-NH2 derivatives to the mu- and delta-opioid receptors, and to the anti beta-endorphin monoclonal antibody (clone 3E7), indicated any change at the Leu5 had little effect on the binding when compared with modifications to the YGGF sequence. Conversely, cyclo-Leu residues did alter the binding of YGGFLRF-NH2 derivatives when substituted for Leu5. Of these 32 peptidomimetics, three derivatives of 2S,3S-cyclo-Leu had relatively low Ki values for binding to an NPFF receptor. Differences between the outcome of the screens were interpreted in terms of the position of the cyclo-Leu residue in the two sequences. PMID- 9354244 TI - Synthesis and protein kinase C inhibitory activities of balanol analogues with modification of 4-hydroxybenzamido moiety. AB - A series of racemic balanol analogues with modification of the benzamido moiety of balanol have been synthesized and evaluated for their inhibitory activities against human protein kinase C isozymes (PKC-alpha, -beta I, -beta II, -gamma, delta, -epsilon, and -eta). The structural modification includes replacement of the 4-hydroxyphenyl group with variously substituted phenyl rings, substitution of the amide linkage with a sulfonamide or an ester, and replacement of the 4 hydroxyphenyl substructure with a hydroxyl substituted indole or a hydroxybenzyl group. in general, these analogues were found to be less potent than balanol, but a number of analogues were identified with improved isozyme selectivity. The structure-activity relationship studies of these analogues also indicated that (1) the optimal general PKC inhibition requires a free 4-hydroxyl group in the benzamido portion of the molecule, (2) the amide linkage of the benzamido moiety is important for PKC inhibition, and (3) the conformation associated with the benzamido moiety seems to have a profound effect on PKC inhibition. The requirement of a free 4-hydroxyl group in conjunction with an appropriate conformation of the benzamido moiety for optimal PKC inhibition suggests that the 4-hydroxyphenyl group may be involved in a specific inhibitor-enzyme interaction important for PKC inhibition. PMID- 9354245 TI - Embodying a stable alpha-helical protein structure through efficient chemical ligation via thioether formation. AB - A new approach was developed to embody the alpha-helical protein structure having an arbitrary combination and arrangement of helices by the successive ligation of a haloacetyl peptide segment with a cysteinyl peptide. A four-helix-bundle protein was efficiently constructed by the repetitive ligation of alpha-helical peptide segments. The use of HPLC-purified unprotected peptide segments facilitated the purification of the intermediates to afford the highly homogeneous desired protein. The use of the bromoacetyl moiety and the chloroacetyl moiety for the ligation was judged to make no difference in practice. A trial of introducing an additional intramolecular disulfide cross link was also examined. The resulting protein showed high stability in the chaotropic and thermal denaturation and in enzymatic degradation. PMID- 9354247 TI - Admission examination factors predicting cognitive improvement during acute brain injury rehabilitation. AB - In this study 46 brain-injured patients admitted to a brain injury inpatient rehabilitation programme after acute care were evaluated at the bedside with a comprehensive mental status examination and physical evaluation. Using multiple regression analysis, elements of the cognitive and physical examinations were studied to determine if any of these items, individually or collectively, had predictive value for discharge functional status. A principal-components analysis identified meaningful clusters of items. Factor I, Simple Cognitive Operations, consisted of word repetition, naming and comprehension of simple commands. Factor IV, Higher Cognitive Operations, consisted of interpretation of similarities and reverse digit span. Other items, including elements of the physical examination such as eye movements, hearing and ambulatory function, were also clustered into factors. Stepwise multiple regression analysis led to a conclusion that only factor IV was a significant predictor of Discharge Rancho Level, with the predictive power of factor I and Admission Rancho Level subsumed by factor IV. This model, using a few items from a bedside mental status examination, accounted for 27% of the variance in Discharge Rancho Level in patients admitted to a brain injury rehabilitation unit after injury. We conclude that a brief mental status examination in a rehabilitation inpatient setting can be useful in predicting outcome in patients with brain injuries. More research is needed to identify additional test items that will add prognostic power to the initial evaluation of patients with brain injuries admitted to a rehabilitation unit. PMID- 9354246 TI - Child and adolescent traumatic brain injury: psychiatric findings from a paediatric outpatient specialty clinic. AB - A record review focused on children and adolescents, with a history of traumatic brain injury, who were consecutively admitted to a brain injury clinic in which all new patients are psychiatrically evaluated. The development of a 'novel' psychiatric disorder (not present before injury) occurred in 76% (38/50) of the cohort and was correlated significantly with family psychiatric history and family function, but not with severity of injury, preinjury psychiatric status, intellectual/educational functioning, or socioeconomic status. Psychiatric consultation is often necessary in this paediatric population even though much of the psychopathology, particularly following mild injury, may not be directly related to brain trauma. PMID- 9354248 TI - The Sensory Modality Assessment Rehabilitation Technique--a tool for assessment and treatment of patients with severe brain injury in a vegetative state. AB - An analysis of data from 30 subjects diagnosed as being in vegetative state (VS) on admission to a specialized Brain Injury Unit was carried out. Rancho Level ratings given by the referring physician were compared with those of the units occupational therapists (OT). Scores were obtained from the Sensory Modality Assessment Technique (SMART) and the Western Neuro Sensory Stimulation Profile (WNSSP) on admission and at 2 monthly intervals and converted to Rancho Level ratings to allow comparison. The comparison of the assessments within one week of admission showed agreement between Rancho Level scores derived from the WNSSP and those from the referring physicians. The Rancho scores derived from the SMART were significantly different from the physicians' and the WNSSP (P < 0.01), with the SMART rating the patient at a higher level of cognitive functioning. Although all 30 subjects were diagnosed as VS on admission, the SMART assessed six subjects not to be in VS within 2 to 4 months from admission and this was established at least 6 weeks earlier than the comparable conclusion from the WNSSP in four subjects. This initial validation study shows that the SMART is a useful tool in discriminating awareness and more sensitive at detecting the higher cognitive functions than both the WNSPP and referring physician, thus indicating the need to conduct a specifically designed prospective study to validate and further evaluate the SMART. PMID- 9354249 TI - Neuropsychological function in restrained versus unrestrained motor vehicle occupants who suffer closed head injury. AB - It is known that using seatbelts reduces the incidence and severity of closed head injury (CHI) from motor vehicle crashes. One would expect unrestrained occupants in motor vehicle crashes to suffer more severe CHIs than restrained occupants, as reflected by Glasgow Coma Scale (GCS) scores. One might also expect an increased risk of focal injury due to contact forces in unrestrained occupants. The purpose of this study was to test the hypothesis that failure to use seatbelts results in increased severity of neuropsychological sequelae, even with GCS controlled. We also examined the impact of demographic variables on seatbelt use. Subjects included patients admitted to a hospital trauma service who were suspected of having suffered CHI. All patients completed neuropsychological testing, which was entered into a data base along with demographic and clinical information. People who had documented use of seatbelt restraints were compared with those who were unrestrained. Results confirmed that certain demographic variables are associated with the use of seatbelts. Results also suggested that failure to use seatbelt restraints is associated with more severe impairment on tests that are sensitive to frontal lobe dysfunction. PMID- 9354250 TI - Anoxic encephalopathy: a neurobehavioural study in rehabilitation. AB - This study examined an operational conditioning paradigm designed to train procedural learning channels in a patient with anoxic encephalopathy. Prior to treatment the patient had been incontinent to bowel and bladder incontinent, minimally participant in daily activities and aggressive toward staff and residents for two years. Previous rehabilitation attempts had been unsuccessful in changing these behaviours. Based on the literature, it was hypothesized that through operant conditioning the patient could be trained on three behaviours: 1) bowel and bladder continence 2) activity level and 3) level of aggression. Results indicated that at a 4 month interval following the initial training, the patient had improved in all areas of functioning. PMID- 9354251 TI - The use of noncontingent reinforcement and contingent restraint to reduce physical aggression and self-injurious behaviour in a traumatically brain injured adult. AB - Many different intervention programmes for reducing undesirable behaviour with people with traumatic brain injury (TBI) have been investigated in recent years. The purpose of this study was to examine the potential of using noncontingent reinforcement (NCR) in combination with contingent restraint to reduce severe behaviour. The subject (E.L.) was a 40-year-old male with TBI admitted to a rehabilitation long-term care programme. E.L. had a history of physical aggression (PA) and self-injurious behaviour (SIB). Assessment conditions included a descriptive analysis, response scatterplot and Self-Injury Trauma (SIT) Scale. Attention was identified as the maintaining positive reinforcement for PA and SIB. Treatment conditions were compared using a reversal (ABAB) design. Attention (NCR) was delivered on a fixed-time schedule that was not dependent on the subject's behaviour. Contingent restraint was applied when E.L. exhibited PA or SIB that was dangerous to himself or others. During treatment, PA occurred over 4 times less often and SIB over 2.5 times less often. Results demonstrated that PA and SIB were sensitive to NCR. NCR can be an effective procedure for reducing severe behaviour maintained by socially-mediated positive reinforcement. PMID- 9354252 TI - Use of electrical stimulation in brain-injured patients: a case report. AB - After failure of other therapeutic measures, electrical stimulation was applied to promote gait rehabilitation in a patient with severe brain injury and complete left hemiplegia. The favourable results reported in the literature were confirmed. Despite the long interval between injury and institution of electrical stimulation, independent ambulation was quickly restored. PMID- 9354254 TI - Buccofacial apraxia and left cerebral haemorrhage. AB - We examined 33 patients with left intracerebral haemorrhage to investigate the relations to buccofacial apraxia (BFA). BFA was present in 18 cases at 1 month and disappeared in 3 cases and persisted in 15 cases at 6 month from the onset. The existence of BFA seems to be partially dependent the haematoma volume which may cause the organic damage in lenticulate nucleus, insula, posterior limb or internal capsule and anterior portion of paraventricle white matter. All patients with BFA had aphasia (Broca 6, Wernicke 1, Total 10, Amnestic 1). Aphasia of the patients with transient BFA improved as well as the patients without BFA after BFA disappeared. We suspected that improvement or aphasia might be affected by BFA. PMID- 9354253 TI - Local histamine release increases leukocyte rolling in the cerebral microcirculation of the mouse. AB - Histamine-mediated induction of leukocyte rolling and adhesion in the cerebral microcirculation was examined in two inbred strains of mice (SJL/J and BALB/c). A cranial window was surgically prepared for the visualization of the cerebral microcirculation using intra-vital microscopy. Leukocyte rolling and adhesion to pial venular walls were assessed during off-line video playback analyses. The surgical preparation of the cranial windows was found to trigger 'spontaneous' leukocyte rolling, and this was attributed to disruption of dural mast cells and localized release of vasoactive histamine. This spontaneous leukocyte rolling was observed only in the SJL/J strain of mice, and could be prevented by presurgical treatment with the mast cell stabilizer sodium cromoglycate. BALB/c mice did not show 'spontaneous' leukocyte rolling or adhesion; this strain is known to have low numbers of CNS-associated mast cells. Exogenous histamine, applied topically to the cerebral microcirculation via the cranial window in mice pretreated with sodium cromoglycate, produced significant dose-dependent increases in leukocyte rolling and adhesion to pial venules in SJL/J mice, but not in BALB/c mice. Diphenhydramine (H1 receptor antagonist), but not cimetidine (H2 receptor antagonist), abolished both 'spontaneous' and histamine-induced leukocyte rolling. Anti-P-selectin antibody was found efficiently to block both spontaneous and histamine-induced increases in leukocyte rolling, but not leukocyte adhesion. PMID- 9354255 TI - 'Postconcussive' symptoms in persons with chronic pain. AB - The purpose of this study was to examine the base rate of cognitive and neurobehavioural complaints in patients with chronic pain (N = 170) who had not sustained a head injury. The patients completed a packet of questionnaires that contained numerous questions regarding physical, cognitive, and psychological symptoms. The 'postconcussive-like' symptoms were selected and analysed. Specific symptom endorsement rates ranged from 5% to 76.5%. Disturbed sleep, fatigue, and irritability were reported by the majority of chronic pain patients. Cognitive complaints relating to forgetfulness (29%), difficulty maintaining attention (18%), and difficulty with concentration or thinking (16.5%) were endorsed by a significant minority of patients. Most patients (80.6%) endorsed three or more symptoms from Category C of the DSM-IV Postconcussional Disorder research criteria. This study further illustrates that postconcussive-like symptoms are not unique sequelae of mild head injury, and the presence of chronic pain should be considered when interpreting patients' physical, cognitive, and psychological complaints following closed head injury. PMID- 9354256 TI - Response bias in plaintiffs' histories. AB - This study investigated response bias in self-reported history of factors relevant to the assessment of traumatic brain injury, toxic brain injury and related emotional distress. Response bias refers to systematic error in self report data. A total of 446 subjects (comprising 131 litigating and 315 non litigating adults from five locations in the United States) completed a symptom questionnaire. Data were obtained from university faculty and students, from patients in clinics specializing in physiatry neurology, and family medicine, and from plaintiffs undergoing forensic neuropsychological evaluations. Comparisons were made for litigant and non litigant ratings of their past and current cognitive and emotional functioning, including life in general, ability to concentrate, memory, depression, anxiety, alcohol, drugs, ability to work or attend school, irritability, headaches, confusion, self-esteem, and fatigue. Although there is no basis for hypothesizing plaintiffs to be healthier than the general population, plaintiffs rated their pre-injury functioning superior to non plaintiffs. These findings suggest that response biases need to be taken into account by forensic examiners when relying on litigants' self-reports of pre injury status. PMID- 9354258 TI - Perseveration and wandering as a predictor variable after brain injury. AB - Perseveration and wandering are well recognized sequelae of traumatic brain injury. They are believed to arise as a constellation of behaviours from lesions involving the frontal, temporal, or parietal lobes or subcortical motor regions. Attention and memory deficits are believed to contribute to the presence of perseveration and wandering. Patients who perseverate and/or wander after brain injury tend to have poorer outcomes and slower rates of progress. This prospective study is based on 32 consecutive admissions to a brain injury rehabilitation unit over a 4-month period. For each patient data was collected on perseverative behaviours, wandering, Functional Independence Measure, total number of rehabilitative days, supervision requirement at discharge and discharge disposition. Perseveration and wandering were assessed using the Agitated Behaviour Scale and clinical observations of the primary physician, physical and occupational therapist and speech-language pathologist. Correlations between perseverative and wandering behaviours were made with the variables identified. PMID- 9354257 TI - Access and organization of goal-derived categories after traumatic brain injury. AB - This study examined the access and organization of goal-derived categories in semantic memory with a group of chronic traumatic brain injured TBI adults and a group of age and gender-matched neurologically-intact controls. Goal-derived categories are developed by individuals for use in specialized contexts to achieve a goal, such as 'things to take on a camping trip.' Categories were presented to subjects in two task contexts: category verification and exemplar generation. Overall, the TBI subjects were able to accurately identify and organize category exemplars within particular categories. Interestingly, the TBI subjects produced significantly more total responses than the neurologically intact subjects on exemplar generation; however, a high percentage of their responses (one-third) were inaccurate, consisting of out-of-set responses and repetitions. These findings suggest that difficulties in retrieval may exist in the presence of relatively intact access and organization of goal-derived category structure. The results are discussed relative to deficits in the executive control of verifying goal-directed behaviour and incomplete category representation. PMID- 9354259 TI - Relationship of family schema to family adaptation to brain injury. AB - The relationship between family schema and family adaptation to brain injury was investigated. Participants were 87 primary caregivers of persons with brain injuries in Wisconsin who are part of a comprehensive, longitudinal study of family adaptation to having a member with a brain injury. Stepwise multiple regression of variables measuring family schema on family adaptation indicated that manageability and meaningfulness were predictive of family adaptation. Thus the hypothesis that family adaptation can be predicted from variables measuring family schema was supported. Family intervention and research implications are discussed. PMID- 9354260 TI - Use of carbidopa-levodopa in a patient with hydrocephalus and frozen movement. AB - While movement disorders are frequently encountered after brain injuries, and may create a host of complicated problems for the clinician, only a few cases of Parkinsonism associated with hydrocephalus have ever been described in the literature. Parkinsonism-like syndrome complicating hydrocephalus is a rare disorder, especially when associated with nontumoral aqueductal stenosis. Yet as this case report discusses, hdyrocephalus-induced Parkinsonism may be responsive to levodopa-carbidopa administration. This report describes a perplexing case of persistent akinesis following corrective surgery for aqueductal stenosis and the subsequent response to levodopa-carbidopa administration. We present the case of a 28-year-old male with a history of non-tumoral aqueductal stenosis diagnosed at age 12. As a child, he underwent a ventriculo-peritoneal shunt placement for obstructive hydrocephalus followed by multiple shunt revisions over the next several years. Sixteen years after his initial shunt placement, the patient presented with a decline in mental status. A third ventriculocisternostomy was performed rather than another shunt revision. Following surgery, the patient remained obtunded, and displayed profound hypokinesis, best described as freezing in movement. Upon admission to a rehabilitation unit 2 weeks later, he had made only minimal progress. A SPECT (single-photon emission computed tomography) brain scan revealed decreased basal ganglia perfusion. Levodopa/carbidopa therapy was initiated and within 2 weeks, the patient showed improvement in speed of movement, facial expression and verbal output. Eight weeks later, the patient could independently complete his basic activities of daily living and demonstrated little, if any, disordered movement. This report illustrates how dopaminergic agents may be useful in cases of hypokinesis following corrective surgery for aqueductal stenosis. SPECT may further aid in the diagnosis and management of Parkinsonism-like syndromes in brain injuries. PMID- 9354261 TI - Protocols for the vegetative state. AB - The diagnosis of the vegetative state, the fate worse than death, brings with it a nihilistic approach to the patient, untold misery to the family, emotional turmoil to medical and nursing staff who work in brain injury units and enormous economic cost to the country. There are no protocols yet devised for the vegetative state. The diagnosis has been shown to be incorrectly applied in a significant number of patients. The ethics of the situation needs full exploration. This paper explores the practical aspects of the diagnosis and management of the vegetative state and present methods of resolving problems associated with them. PMID- 9354262 TI - Sequence analysis of the gene encoding the capsid protein of the Snow Mountain human calicivirus. AB - Snow Mountain virus (SMV) is the reference strain for serotype 3 as determined by immune electron microscopy of the human caliciviruses that are associated with epidemic gastroenteritis. In order to establish the genetic relationship of its capsid protein with those from other human caliciviriuses, the sequence of the open reading frame 2 (ORF2) encoding the SMV capsid protein was determined. The SMV ORF2 sequence was 1626 nucleotides in length and the deduced protein of 542 amino acids had a calculated molecular weight of 59.2 kD. The SMV capsid sequence showed approximately 48 and 77% amino acid sequence identity with the capsid proteins of the Norwalk (serotype 1) and Hawaii (serotype 2) human calicivirus reference strains, respectively, a finding consistent with its serotypic distinctiveness. Furthermore, the predicted amino acid sequence of the SMV capsid was found to share highest sequence identity (98%) with the Melksham human calicivirus in database searches. PMID- 9354263 TI - Genomic localization and nucleotide sequence of a lef-8 gene of the Spodoptera littoralis nucleopolyhedrovirus. AB - A gene similar to lef-8 of the Autographa californica (Ac) nucleopolyhedrovirus (MNPV) was identified in the Spodoptera littoralis (Spli) MNPV. The SpliMNPV lef 8-like gene was localized on the genomic map between 26.9 and 29 map units and is flanked by a chitinase gene and p47 gene. This gene arrangement differs from that of similar genes in the AcMNPV genome, where the lef-8 gene is located about 62 kbp from the chitinase gene and about 7 kbp from the p47 gene. Sequence analyses of the lef-8 gene revealed an ORF of 2730 nucleotides, predicted to encode a protein with Mr 104876. The putative protein is 60.9% identical to the AcMNPV LEF 8 and contains an identical sequence of a conserved motif of DNA-dependent RNA polymerases. Sequences downstream of the lef-8 gene contain two sequence repeats which resemble a repeated motif of the SpliMNPV enhancer element and other repetitive sequences from the viral genome. PMID- 9354264 TI - Deletion analysis of both the long terminal repeat and the internal promoters of the human foamy virus. AB - Deletion analyses of the long terminal repeat (LTR) and internal promoters (IP) of human foamy virus (HFV) showed that a negative acting element resides in the U5 region of the 5' LTR reducing reporter gene expression tenfold. The basal activity of the IP was higher than that obtained with LTR promoter constructs and strongly elevated in permissive BHK-21 cells whereas semi-permissive COS-7 cells showed low basal activity. Since the basal activity of the IP is critical for initiating HFV gene expression by providing Bel 1 transactivator early after infection, the basal activity of the IP may be the crucial factor that contributes to whether cells are permissive for HFV infection or not. Deletion mutagenesis allowed to define the minimal IP region. A region strongly transactivated by Bel 1 extends from -136 to +58 relative to the cap site of the IP. The major Bel 1 response element of the IP required for transactivation is located upstream of the cap site between -136 and -88 relative to the internal cap site. A DNA fragment reported to be protected by recombinant Bel 1 was deleted with marginal reduction of Bel 1 transactivation. HFV gene expression directed by the IP and LTR promoters is thus multiply regulated by positive and negative acting response elements in cis and their binding partners in trans. PMID- 9354265 TI - The genomic 5' terminus of Manchester calicivirus. AB - An enteric calicivirus showing the classic cup-shaped surface morphology was identified in a stool sample obtained from a child with symptoms of acute gastroenteritis (Portishead virus, PHV). Genomic RNA was extracted directly from the PHV stool sample and amplified by RT-PCR using primers based on the Manchester isolate of HuCV. The 3' terminus of the cDNA was defined by homopolymer tailing with dATP and revealed an additional 165 nucleotides suggesting that the previously determined Manchester HuCV (MV) genome sequence was incomplete. Homopolymer tailing of MV cDNA primed using sequence data from the 5' terminus of PHV allowed extension of the MV genome by a further 165 nucleotides thereby increasing the overall genome length to 7431 nucleotides and resulting in an additional 72 amino acids at the N-terminus of the polyprotein. A conserved sequence motif typical of other caliciviruses was also identified at the extreme 5'-terminus of the genome. PMID- 9354266 TI - PCR analysis of human telomeric repeats present on HHV-6A viral strains. AB - Human herpesvirus 6 (HHV-6) presents a perfect tandem array of human telomeric repeats (TRS) at both identical direct repeats (DR). Several researchers have reported a different TRS copy number by sequence analysis of HHV-6 DR's cloned fragments so it has been hypothesized that number of TRS is unstable. By PCR we show that the TRS copy number of U1102 HHV-6 variant A strains is stable during viral cultivation in cell lines and each HHV-6 variant A strain, detected in pathologic specimens, is characterized by a specific TRS copy number. PMID- 9354267 TI - A ribozyme targeted to cleave the polymerase gene sequences of different foot-and mouth disease virus (FMDV) serotypes. AB - Vaccinations against foot-and-mouth disease virus (FMDV) has dramatically reduced the number of disease outbreaks. Nevertheless, there are still many outbreaks in different regions around the world. In an effort to find new ways to control the disease, ribozymes able to cleave FMDV were designed and tested. In this work we tested the ability of FRZ4, a ribozyme targeted to the viral polymerase gene, to cleave polymerase sequences of several FMDV. Homology analysis was used to choose target sequences which consist of two conserved GUC which lie 15 bases apart and, their flanking sequences. These were the basis for the FRZ4 ribozyme gene sequence that contains two catalytic domains. We show that polymerase sequences from A, Asia 1, C and two different O1 Israeli isolates could be specifically cleaved by FRZ4. It is suggested that FRZ4 can cleave polymerase gene sequences from any FMDV serotype. PMID- 9354269 TI - Herpesvirus of turkeys homologue of HSV VP16 is structurally related to varicella zoster virus trans-inducing protein encoded by ORF 10. AB - Expression of the immediate-early genes of alpha-herpesviruses is stimulated by a family of trans-inducing factors represented by VP16 of HSV-1 and ORF10 gene product of VZV. We have identified and determined the nucleotide sequence of the UL48 gene encoding the herpesvirus of turkeys (HVT) homologue of HSV VP16. The gene maps to the BamHI-J fragment and appears to be expressed in a form of bicistronic transcript together with UL49. The deduced amino acid sequence of the protein encoded by HVT UL48 gene shows 55% identity with MDV UL48 gene product. Like the majority of related proteins in other alpha-herpesviruses, the protein encoded by HVT UL48 gene lacks the acidic C-terminal tail, known to possess the transactivation capacity of HSV VP16. Hydrophobic cluster analysis has revealed that its predicted domain composition is closely related to the transactivator protein encoded by ORF10 of VZV. However, the putative amino-terminal activation domain of the HVT homologue of HSV VP16 does not contain a typical horseshoe-like hydrophobic cluster found in other alpha-herpesvirus homologues, suggesting either that it acts as a transactivator via a different activation domain or that its transactivation potential is diminished. PMID- 9354268 TI - Sequence analysis demonstrates that VP6, NSP1 and NSP4 genes of Indian neonatal rotavirus strain 116E are of human origin. AB - We have sequenced the genes encoding the inner capsid protein VP6 and the nonstructural proteins NSP1 and NSP4 of the Indian neonatal serotype P8[11]G9 human/bovine reassortant candidate vaccine rotavirus strain 116E. These three genes share a high degree of sequence and deduced amino acid homology with human prototype strain Wa. Our results confirm and extend those of previous RNA-RNA hybridization studies which suggested that these genes are of human origin, and will facilitate examination of the host immune response to 116E induced by natural infection and vaccination. PMID- 9354270 TI - Evolutionary analysis of influenza C virus M genes. AB - The previous study of the 25 hemagglutinin-esterase (HE) glycoprotein genes of influenza C viruses identified four discrete lineages represented by C/Yamagata/26/81, C/Aichi/1/81, C/Aomori/74 and C/Mississippi/80, respectively. Here we compared the M gene sequence among the 24 viruses isolated between 1964 and 1991. A phylogenetic analysis showed that these genes have evolved into three distinct lineages. Lineage I included most of viruses with the HE genes of C/Yamagata/26/81-related lineage. The predominant members of lineage II were viruses having the HE genes of either C/Aichi/1/81- or C/Mississippi/80-related lineage. Lineage III contained only C/Aomori/74. Phylogenetic positions of several strains (C/Yamagata/64, C/Kanagawa/1/76, C/Miyagi/77 and C/Nara/1/85) were different between the M and HE gene trees, suggesting that they are reassortants. Furthermore, phylogenetic relationships between C/Mississippi/80 like and C/Aichi/1/81-like viruses were much closer for the M gene than the HE gene, raising the possibility that these two virus groups are genetically related by a reassortment event. Nucleotide changes in the M genes occurred at about 7% positions with a uniform distribution throughout the molecules. However, the predicted amino acid sequence of the matrix protein (M1) was conserved almost completely among the isolates analyzed. The amino acid sequence of the second protein (CM2) encoded by M gene was also highly conserved, but was more divergent than the M1 protein sequence, suggesting that the two M gene products are evolving differently in response to selective pressures or structural and functional constraints. PMID- 9354271 TI - Sequences of the three coat protein genes of a Malaysian isolate of rice tungro spherical virus reveal a close relationship to the Philippine isolate. AB - Coat protein genes CP1, CP2 and CP3 of an isolate (MaP1) of rice tungro spherical virus (RTSV) from Malaysia were isolated, cloned and sequenced. Comparative analysis indicated that MaP1 isolate is closely related to the Philippine isolate. PMID- 9354272 TI - Chicken rotavirus Ch-1 shows a second type of avian VP6 gene. AB - VP6 protein from chicken rotavirus Ch-1 showed more than 13% amino acid differences in comparison with pigeon and turkey VP6 sequences. This difference is greater than that observed between subgroup I and II mammalian rotavirus VP6 sequences. Phylogenetic tree analysis demonstrated that RV Ch-1 VP6 is not a link between avian and mammalian VP6 sequences. RV Ch-1 showed variant aa in 17 positions which were otherwise absolutely conserved in mammalian and avian group A RVs. The 17 replacements were scattered through the entire VP6 sequence except the C-terminal part implicated in the assembly of subviral particles. In RV Ch-1 the proline residue 309, reported to be critical for the trimerization of VP6, was replaced by leucine, but VP6 trimers were still observed. The sequence and hydrophilicity analysis of avian RV VP6 do not explain the anomalous electrophoretic migration behavior of avian VP6 proteins. PMID- 9354273 TI - Nucleotide sequences of the 3' terminal region of onion yellow dwarf virus isolates from Allium plants in Japan. AB - The 2032 nucleotide sequence of the 3' terminal region of onion yellow dwarf virus (OYDV) isolated from Allium wakegi, bearing the genes for viral coat protein (CP) and a truncated RNA-dependent RNA polymerase, has been determined. Respective homologies of the nucleotide sequence in the corresponding region and the deduced amino acid sequence of CP with the equivalents of leek yellow stripe virus (LYSV) from garlic were 68.0 and 59.3%. Variation in the nucleotide sequence is concentrated in the boundary region between the putative RNA dependent RNA polymerase gene and the CP gene as well as in the 3' noncoding region. These sequence divergencies, including the deletion of 79 nucleotides, resulted both in alterations to the amino acid sequence and the absence of 28 amino acid residues in the amino terminal region of OYDV CP in comparison with LYSV CP. In addition, the length of the 3' noncoding sequence of OYDV was one third that of LYSV. Comparison of the 3' terminal 1197 nucleotides sequence of OYDV with sequences of the respective cDNAs cloned by RT-PCR directly from the total RNA of infected Allium plants that included two varieties of A. fistulosum, "Wakenegi" and "Shimonita-negi", and A. chinense, showed 90.7% overall identities, even though they have long been cultivated in locally restricted area in Japan. These findings appear to suggest that a single strain of OYDV invaded Japanese Allium plants long ago and spread throughout them. PMID- 9354274 TI - Natural spillover of a distinctly Canidae-associated biotype of rabies virus into an expanded wildlife host range in southern Africa. AB - Rabies enzootics in southern Africa are associated with two genetically distinct groups of viruses, thought to be adapted to two different sets of host species. The virus groups are referred to as the canid biotype (infecting carnivores of the family Canidae) and the viverrid biotype (infecting carnivores of the subfamily Viverrinae). Cross- or spillover infections of one biotype into the host range of the other are thought to occur from time to time. However, very little is known about this phenomenon and its role in the epidemiology of rabies in southern Africa. We have investigated spillover by monoclonal antibody and nucleic acid sequence analysis of a wide range of virus isolates. Although the inverse had been documented, this report constitutes the first evidence of spillover of canid biotype viruses into viverrid hosts. Our genetic analysis was focused specifically on the G-L intergenic region of the virus genome, thought to be a remnant or pseudogene and it was indicated that, with respect to this region of the genome, spillover does not influence the phylogeny of virus isolates. PMID- 9354275 TI - Cloning and sequence analysis of a non-structural gene of an aquareovirus. AB - The nucleotide and deduced amino acid sequence of genome segment 11 encoding a nonstructural protein of an aquareovirus strain SBR have been determined. Nucleotide sequence analysis showed that the genome segment 11 of SBR virus is 780 nucleotides long and contains a major open reading frame that codes for a polypeptide of 236 amino acids with a predicted molecular weight of 25,504 Da. The second reading frame of genome segment 11 was 480 nucleotides long and codes for a polypeptide of 145 with a predicted molecular weight of 15,715 Da. The genome segment 11 contains 24 nontranslated nucleotides at the 5'-end and 48 nontranslated nucleotides at the 3'-end. This gene codes for two nonstructural polypeptides NS29 and NS15. Comparison of the deduced amino acid sequence of this gene with the published sequences of other members of the family Reoviridae indicated no sequence relatedness. PMID- 9354276 TI - Characterization of the early region 4 of porcine adenovirus type 3. AB - The nucleotide sequence of a 3028 bp DNA segment, located between map co ordinates 100 and 92 in the genome of porcine adenovirus type 3 (PAV-3), was determined. The segment includes the entire early region 4 (E-4) and the right inverted terminal repeat sequences. There were two TATA boxes and one canonical polyadenylation signal on the 1 strand. Homology searches of the GenBank data base for the predicted amino acid sequences revealed that, of the eight open reading frames (ORFs) on the 1 strand, and four ORFs on the r strand, only ORF 8 on the 1 strand showed homology with the 34 kDa E-4 protein of human adenovirus types 2, 12 and 34. Northern blot analysis showed that transcription from the E-4 region of PAV-3 began 4 h after infection, peaked at 8 h and declined after 10 h, before DNA replication began 16 h after infection. The E-4 region of PAV-3 was further characterized by 5' and 3' end mapping of the transcription unit. PMID- 9354277 TI - Ground reaction forces in horses, assessed from hoof wall deformation using artificial neural networks. AB - An artificial neural network (ANN) was developed to investigate whether hoof wall deformation could be used to determine ground reaction forces (GRF) in horses. The ANN was taught this relationship under certain conditions and was able to generalise this knowledge to conditions for which it was not trained before. To acquire data to train and test the ANN, a horse was equipped with strain gauges attached to the dorsal, lateral and medial parts of the hoof to assess hoof wall deformation. A force plate was used to measure the GRFs. Both hoof wall deformation and GRF were recorded simultaneously at different speeds, gaits, surfaces and loads. An ANN was trained with some of these data, and subsequently provided with strain gauge recordings of strides, not used for training. By comparing the GRF calculated by the ANN based on the hoof wall deformation with that recorded simultaneously by the force plate, the generalisability of the ANN was determined. It was found that an ANN is capable of 'learning' the relationship between hoof wall deformation and GRF, and to generalise it to a wide range of new conditions. This technique enables assessment of GRF under difficult conditions, such as on a treadmill or on surfaces where a force plate cannot be employed. PMID- 9354278 TI - Impact during equine locomotion: techniques for measurement and analysis. AB - Impact is implicated in the development of several types of musculoskeletal injury in the horse. Characterisation of impact experienced during strenuous exercise is an important first step towards understanding the mechanism for injury. Measurement and analysis of large, short duration impacts is difficult. The measurement system must be able to record transient peaks and high frequencies accurately. The analysis technique must be able to characterise the impact signal in time and frequency. This paper presents a measurement system and analysis technique for the characterisation of large impacts. A piezo-electric accelerometer was securely mounted on the dorsal surface of the horses hoof. Saddle mounted charge amplifiers and a 20 m coaxial cable transferred these data to a PC based logging system. Data were down-loaded onto a UNIX workstation and analysed using a proprietary statistics package. The values of parameters calculated from the time series data were comparable to those of other authors. A wavelet decomposition showed that the frequency profile of the signal changed with time. While most spectral energy was seen at impact, a significant amount of energy was contained in the signal immediately following impact. Over 99% of this energy was contained in frequencies less than 1250 Hz. The sampling rate and the frequency response of a measurement system for recording impact should be chosen carefully to prevent loss or corruption of data. Time scale analysis using a wavelet decomposition is a powerful technique which can be used to characterise impact data. The use of contour plots provides a highly visual representation of the time and frequency localisation of power during impact. PMID- 9354279 TI - Noninvasive photoelastic method to show distribution of strain in the hoof wall of a living horse. AB - A photoelastic method used for materials testing in industry was adapted to show the distribution of strain through the hoof wall in the living horse. Strain was a change in length per unit length in the material of the loaded hoof wall compared with the unloaded condition. Coloured fringes appeared in the photoelastic plastic where there were differences in strain between adjacent sites (strain gradients) in the hoof. Strain distribution was observed in the shod and unshod hoof wall of the front hooves of 6 sound horses with hooves that appeared 'good' to visual inspection, and one unsound horse with hoof cracks. No significant differences in strains were apparent across the hoof walls of the sound horses when the horses were standing normally. Steep strain gradients were apparent in hooves, associated with defects such as cracks, unstable nail holes, and long toes. PMID- 9354280 TI - Image analysis to measure activity index of animals. AB - The objective of the study was to present a method to quantify the behavioural response of animals to their micro-environment by using a camera system and a digitiser board. An algorithm was developed for analysing images and calculating activity, occupied zone and boundary of the animals. The developed method was tested on 3 different applications and animals. In the first application, the behavioural responses of broiler chickens to their thermal environment was measured. In the second application behavioural responses of pigs to their thermal environment were measured. In the third application, the response of water fleas to a chromium pollution was measured using the developed technique. Based on the experimental results, it can be concluded that the developed image analysis technique can be employed to quantify the behavioural responses of the tested animals to their micro-environment, in an easy and accurate way. PMID- 9354281 TI - Radiological assessment of the effects of a full rolling motion shoe during asymmetrical bearing. AB - The authors used a new radiological method to assess asymmetrical articular compression of the interphalangeal joints. This method was based on the measurements of 3 angles obtained on dorsopalmar radiographs. Variations of these angles were studied during experimental asymmetrical bearing on unshod feet. It was concluded that 2 angles were interesting parameters to assess asymmetrical articular compression and to define the position of the phalanx in the horny box. Furthermore, variations of these angles induced by experimental asymmetrical bearing were compared without shoe, with a standard shoe and with an orthopaedic full rolling motion shoe. It was observed that the effects of the asymmetrical bearing were reduced when feet were shod with a full rolling motion shoe. PMID- 9354282 TI - Mechanical properties of pathological equine superficial digital flexor tendons. AB - The objective of this study was to mechanically characterise superficial digital flexor tendon (SDFT) lesions. Eight pathological SDFTs, isolated from 6 adult horses, were tested in traction until rupture (at 1 mm/s). The stresses and strains simultaneously undergone by each of the 7 segments of a tendon were determined throughout the test, and the modulus of elasticity of each segment was evaluated from the segmental stress-strain curve thus obtained. These mechanical data were compared to those obtained on 10 normal SDFTs. After the test, the tendinous segments were submitted to a histological examination in order to characterise the tissues. Three lesional categories (I to III, of increasing maturity), as well as the normal tendinous tissue, were defined and assessed quantitatively according to their extent in the histological sections. The most recent and severe lesions (categories I and II) were correlated with a large degree of hypertrophy (often above 200%) of the corresponding segments, with a resulting decrease in the stress at tendon rupture, and a slight decrease in the strain at tendon rupture in spite of a low modulus of elasticity (low stiffness). In contrast, the adjacent areas, less or not injured, underwent compensatory strains. This relative overstraining was especially critical with category III tissue, often present in the transitional areas between sound and severely injured segments. Here the modulus of elasticity was low whereas the hypertrophy was only slight. Therefore, the corresponding segments seemed to be the most fragile sites, and those most predisposed to recurring injury, in an injured SDFT. PMID- 9354283 TI - Recovery of equine forelimb function after desmotomy of the accessory ligament of the deep digital flexor tendon. AB - The recovery process of the equine locomotor system after desmotomy of the accessory ligament of the deep digital flexor tendon (AL-DDFT) was investigated by studying the movement patterns and joint moments in 6 horses before and 10 days and 6 months following surgery. Using a modified CODA-3 system the joint angles and angular velocities of the lower limb were assessed in the operated forelimb as before the operation. Simultaneously ground reaction forces were measured and joint moments calculated. At 10 days and 6 months after the operation the carpal joint started to bend earlier in the stance phase. At that instant, the fetlock joint was more extended and displayed a higher angular velocity. The moment of the coffin joint was significantly decreased 10 days after desmotomy. After 6 months it had recovered considerably, but still the shape of the curve was significantly different compared to that before the operation. The fetlock joint moment was not affected, but turned out to be generated for a greater part by the suspensory ligament and the superficial digital flexor 10 days after the operation. Further analysis of these results showed that 6 months after the desmotomy the locomotor system was able to cope with almost similar external moments. To accomplish this, it had adopted a new co ordination pattern during the recovery process. PMID- 9354284 TI - Model for injury to the foreleg of the Thoroughbred racehorse. AB - A discussion is presented of contributing factors to the injury to the foreleg of the Thoroughbred racehorse. The critical part of the step is taken to be the first 10-20 ms after ground contact as the hoof slides forward and stops. Large nonaxial loads associated with the deceleration of the hoof are shown to arise. Results of accelerometer measurements on the hoof of a horse running at racing speed are presented as well as mechanical properties of the racing surface. The mechanical properties of the track surface, the type of shoe, and the degree of fatigue of the horse all work together to determine the level of risk. PMID- 9354285 TI - Joint moments and power in equine gait: a preliminary study. AB - A method is described for the estimation of joint moments of force and power in the equine forelimb using S-VHS video and force platform data. Video and force data were collected for 5 walking trials in a sound Dutch Warmblood horse. The sagittal plane positional and angular data were combined with the vertical and cranio-caudal ground reaction forces to calculate net joint moments of force in the sagittal plane across the carpal, fetlock and coffin joints during the stance phase of the forelimb. The mechanical power was calculated as the product of the netjoint moment and the joint's angular velocity. The carpus demonstrated oscillating periods of energy generation and absorption in early stance against a flexor moment, then an absorption phase at the end of stance as the carpus flexed against an extensor moment. The fetlock absorbed energy in early stance as it flexed against an extensor moment, then showed short periods of positive and negative work in mid-stance. Energy was generated across the fetlock as it extended for push-off. An extensor moment was measured at the coffin joint through most of stance, and this coincided with a phase of energy absorption followed by a longer phase of generation. In terminal stance, a flexor moment was produced and this was accompanied by a period of generation for push-off. PMID- 9354286 TI - Power flow in the equine forelimb. AB - A method is described for the estimation of segmental powers and power flow during the stance phase in the equine forelimb, to demonstrate the sources and paths of energy flow through the limb segments. S-VHS video and force platform data were collected for 5 walking trials in a sound Dutch Warmblood horse. Two camera views were combined using direct linear transformation and the resultant sagittal plane positional and angular data used together with the vertical and cranio-caudal ground reaction forces to calculate moments about the ends of the 4 lowermost segments of the forelimb, and the reaction forces at the segment ends. Power flows were calculated across the proximal and distal ends of each segment and total segmental power computed. During initial and terminal stance, power flowed into the cannon segment proximally, and out distally. For the rest of stance, the flow in the cannon was distal to proximal. At the pastern, power flowed in proximally during initial loading, and out distally. For most of the rest of stance, the pattern of flow was distal to proximal, except for terminal stance, when power flowed in through both ends. The largest effect at the hoof is a loss of energy in terminal stance as power flowed out proximally and into the pastern. The pastern appears to receive most of the energy during loading and pushoff and transfers this energy up the limb during midstance. PMID- 9354287 TI - Components of the total kinetic moment in jumping horses. AB - Thirty horses were filmed with a panning camera operating at 50 frames/s as they jumped over a 1.20 x 1.20 m fence. The markers of 9 joints on the horse and 7 joints on the rider were tracked in 2D with the TrackEye system. The centre of gravity and moment of inertia of each segment were calculated using a geometric algorithm and a cylindric model, respectively. The kinetic moment of each part of the horse was calculated after filtering, and resampling of data. This method showed the relative contribution of each body segment to the body overall rotation during the take-off, jump and landing phases. It was found that the trunk, hindlimbs and head-neck had the greatest influence. The coordination between the motion of the body segments allowed the horse to control its angular speed of rotation over the fence. This remained nearly constant during the airborne phase (120 +/- 5 degrees/s). During the airborne phase, the kinetic moment was constant because its value was equal to the moment of the external forces (722 +/- 125 kg x m2/s). PMID- 9354288 TI - Analysis of the equine jumping technique by accelerometry. AB - The purpose of this study was to demonstrate the relationships between jumping technique and dorsoventral acceleration measured at the sternum. Eight saddle horses of various jumping abilities competed on a selective experimental show jumping course including 14 obstacles. An accelerometric belt fastened onto the thorax continuously measured the dorsoventral acceleration during the course. At each jump, 11 locomotor parameters (acceleration peaks, durations and stride frequency) were obtained from the dorsoventral acceleration-time curves. The type of obstacle significantly influenced the hindlimb acceleration peak at take-off and the landing acceleration peak (P<0.01). The poor jumpers exhibited a higher mean forelimb acceleration peak at take-off, a higher forelimb/hindlimb ratio between peaks of acceleration (F/H), and a lower approach stride frequency than good jumpers. Knocking over an obstacle was significantly associated with a low hindlimb acceleration peak at take-off and a high F/H ratio (P<0.01). In order to observe the continuous changes in the frequency domain of the dorsoventral acceleration during the approach and take-off phase, a Morlet's wavelet analysis was computed for each horse jumping over a series of 3 vertical obstacles. Different patterns of time-frequency images obtained by wavelet analysis were found when the horse either knocked over a vertical obstacle or cleared it. In the latter case, the image pattern showed an instantaneous increase in stride frequency at the end of the approach phase, and a marked energy content in the middle frequency range at take-off. PMID- 9354289 TI - Effect of added weight on landing kinematics in jumping horses. AB - Six event horses jumped a 1.10 m high table fence 4 times under each of 2 conditions; the rider weight condition involved carrying the weight of the rider and saddle (61 kg), whereas the added weight condition included an additional 18 kg weight cloth. Sagittal view, 60 Hz video recordings were analysed using standard methods. Comparisons between the rider weight and added weight conditions using paired t tests (P<0.05) showed a number of significant differences. In the added weight condition the leading forelimb landed closer to the fence, and there were increases in the maximal extension of the fetlock and carpal joints in this limb during the landing phase. In the first departure stride, the stance durations of both hindlimbs increased, and the advanced placement between them was reduced for the added weight condition. The head was significantly further ahead of the vertical in the added weight condition at the instants of ground contact of the TrH, LdH and TrF in the first departure stride. PMID- 9354290 TI - Classification of collected trot, passage and piaffe based on temporal variables. AB - The objective was to determine whether collected trot, passage and piaffe could be distinguished as separate gaits on the basis of temporal variables. Sagittal plane, 60 Hz videotapes of 10 finalists in the dressage competitions at the 1992 Olympic Games were analysed to measure the temporal variables in absolute terms and as percentages of stride duration. Classification was based on analysis of variance, a graphical method and discriminant analysis. Stride duration was sufficient to distinguish collected trot from passage and piaffe in all horses. The analysis of variance showed that the mean values of most variables differed significantly between passage and piaffe. When hindlimb stance percentage was plotted against diagonal advanced placement percentage, some overlap was found between all 3 movements indicating that individual horses could not be classified reliably in this manner. Using hindlimb stance percentage and diagonal advanced placement percentage as input in a discriminant analysis, 80% of the cases were classified correctly, but at least one horse was misclassified in each movement. When the absolute, rather than percentage, values of the 2 variables were used as input in the discriminant analysis, 90% of the cases were correctly classified and the only misclassifications were between passage and piaffe. However, the 2 horses in which piaffe was misclassified as passage were the gold and silver medallists. In general, higher placed horses tended toward longer diagonal advanced placements, especially in collected trot and passage, and shorter hindlimb stance percentages in passage and piaffe. PMID- 9354291 TI - Comparison of the temporal kinematics of the canter pirouette and collected canter. AB - The objectives were to compare the temporal characteristics of canter pirouette strides with collected canter strides in elite dressage horses, and to determine whether the stride kinematics of the canter pirouettes fulfilled the requirements specified in the Federation Equestre Internationale Rules for Dressage Events. Eleven horses were videotaped (60 fields/s) during the individual medal competition at the 1992 Olympic Games. Temporal variables were extracted from the videotapes using standard methods. Two strides were analysed on each of the left and right leads and these were pooled to give mean values for the collected canter and the pirouettes. The pirouettes were completed in 4-9 strides, (mean of 6.4). In the collected canter strides, mean duration of the suspension was 0.013 s. There was no suspension in any of the pirouette strides, instead the stance phases of the leading forelimb and trailing hindlimb overlapped by a mean of 0.163 s. In 9 horses the trailing forelimb contacted the ground before the diagonal leading hindlimb in the collected canter, whereas in the pirouettes the leading hindlimb always made contact before the trailing forelimb (mean dissociation 0.164 s), giving the strides a distinct 4 beat rhythm. Due to increases in advanced placement between the diagonal limb pair and between the 2 forelimbs, the stride duration was longer in the pirouette (0.879 s) than the collected canter (0.629 s). It is concluded that the canter pirouette strides did not maintain the rhythm and timing of the the collected canter strides in any of the 11 horses. PMID- 9354292 TI - Effects of trot quality and collection on the angular velocity in the hindlimbs of riding horses. AB - The angular velocities of the hindlimb angles of 14 horses, including 6 Grand Prix dressage horses, 4 horses judged as good at the trot and 4 horses judged as poor, were analysed. The horse material was the same as previously used by Holmstrom (1994) in studies on conformation and trotting gaits in the Swedish Warmblood riding horse. Four consecutive strides of each horse and the corresponding pace were analysed and mean velocity curves (Xh) for each angle were calculated. Before calculation the data were filtered forwards and backwards with a Butterworth third order filter with a cut off frequency of 60 Hz. During the last 60% of the stance phase there were differences between the horses judged as good and poor at the trot in all the analysed hindlimb angles except the femur inclination. The angular velocity in the hock joint, pelvis inclination and hindlimb pendulation was larger in the good horses. The angular velocity of the hindlimb pendulation decreased with collection in the Grand Prix horses. During parts of the stance phase, there was also a gradual decrease in the femur angular velocity from trot at hand to piaffe. In the hock joint, there was no difference in angular velocity between trot at hand and passage during the last 30%. The higher compression of the hock angle and pelvic angle to the horizontal plane probably reflects a higher compression of the whole hindlimb. It probably contributes to the greater springiness in the movements of good young horses and Grand Prix dressage horses. The results from the present study confirmed the importance of storing elastic strain energy for the quality of the dressage horse gaits. PMID- 9354293 TI - Aspects of postural development in the rat. AB - The activation patterns in the longissimus and gastrocnemius muscles during the development of the mature type of locomotion were studied in rats from the 11th day until maturity. The EMG's of these muscles were recorded and movements were simultaneously recorded on videotape. Results indicate that during locomotion at early ages, the longissimus muscles are activated irregularly. From the 16th day, the bursts in the longissimus muscle become more pronounced. During locomotion, they are activated differentially and often a simultaneous activation of the longissimus and the contralateral gastrocnemius muscle was observed. From the 20th day, bursts in the gastrocnemius muscle during walking coincide with bursts in the ipsilateral longissimus muscle. Our results demonstrate that the mature type of postural control (from the 20th day) develops a few days after the development of the mature type of fluent walking (around the 15th-16th day). PMID- 9354294 TI - Effects of treadmill inclination on kinematics of the trot in Dutch Warmblood horses. AB - To evaluate the effects of uphill trotting on stride characteristics, 6 well trained Dutch Warmblood horses trotted at 4 m/s on a horizontal and on an inclined (6%) treadmill. This was done under 3 different conditions, unloaded, mounted by an experienced 90 kg rider and loaded with 90 kg of lead, to study whether extra weight provoked more or different alterations than the incline per se. In all 3 test situations (unloaded, mounted and lead-loaded), heart rates were significantly higher on the inclined treadmill than on the horizontal treadmill. Stride duration tended to increase on the inclined treadmill. Stance duration increased significantly on a slope, more in the hindlimbs than in the forelimbs. In the unloaded condition, maximal fetlock extension of the forelimb decreased on the incline, whereas maximal fetlock extension of the hindlimb and tarsal range of motion increased significantly on the slope in all 3 conditions. The overall effect was that on an inclined treadmill the hindlimbs seemed to carry more weight (higher maximal fetlock extension), and to provide greater propulsion (higher tarsal flexion and increased pro/retraction). PMID- 9354295 TI - Comparison of stride characteristics in a cantering horse on a flat and inclined treadmill. AB - The purpose of this study was to determine whether there was any difference in the stride characteristics between cantering on a flat or inclined treadmill. Five 2-year-old Thoroughbred horses were cantered on a treadmill at 3 different velocities and at 3 different slopes. The sequence of speeds at each slope was chosen at random and 16 mm cinefilms at 300 frames/s were taken from a lateral view at a distance of 15 m from the treadmill to record the linear and temporal data. On the slope, stride length, stride duration, stance duration and swing duration did not change. However, midstep length increased significantly and the airborne step length decreased significantly as slope increased from a 3% slope to a 8% slope at 12 m/s. The airborne duration increased significantly from a 0% slope to a 3% slope, and inversely decreased significantly from a 3% slope to a 8% slope. The advanced placement (AP) between trailing hindlimb and leading hindlimb (APTH-LH) and between trailing forelimb and leading forelimb (APTF-LF) and the overlap between LH and TF (LH-TF) tended to decrease and the AP between LH and TF (APLH-TF) and the overlap between TH and LH (TH-LH) inversely tended to increase as the slope increased, though these tendencies were not significant. These findings indicated that in slope locomotion, the stride length might be maintained by sacrificing the length of the airborne period as the workload became more intense. PMID- 9354296 TI - Kinematic comparison of the leading and trailing fore- and hindlimbs at the canter. AB - The canter is a 3 beat asymmetrical gait with a difference in timing between left and right limbs. To evaluate intralimb asymmetry at the canter, a group of 24 Dutch Warmbloods was evaluated on a treadmill (7 m/s) using a modified CODA-3 optoelectronic gait analysis system. Thirteen horses cantered in the left lead ('leading limb' group) and 11 in the right lead ('trailing limb' group) during left forelimb recordings, while 11 horses were at the left and 13 were at the right lead during left hindlimb recordings. Kinematic differences between horses from the 'leading limb' and 'trailing limb' group were statistically evaluated at a significance level of P<0.05. Stride, stance and swing duration were similar between the 2 groups. The pelvis rotation, angle of maximal protraction and total range of maximal pro- and retraction were larger in the 'leading limb' group, while the scapula rotation, and the angle of maximal retraction were larger in the 'trailing limb' group. The elbow and hip joints were more flexed at impact, at maximal extension and at maximal flexion of the leading limb, whereas the stifle joint was more extended at impact. Furthermore, the leading tarsal joint was more maximally flexed in stance and swing phase, whereas the carpal joint was more flexed only in the swing phase of the leading limb. However, during the stance phase the maximal fetlock extension of the trailing fore- and hindlimbs were significantly larger. Apparently, horses move at the canter with a more protracted leading limb by more flexing the elbow, carpal, hip and tarsal joints. In the trailing limb, however, the scapula is more rotated, and the tarsal and fetlock joints are more loaded. In conclusion, the difference in interlimb timing between left and right limbs at canter also leads to an asymmetry in intralimb coordination of these limbs. PMID- 9354297 TI - Kinematics and kinetics of the carpus. AB - This study investigated the kinematics and kinetics of the carpus during the stance phase. Five Standardbred horses trotted on a treadmill at 8.9 m/s. The kinematics of the horses were filmed and hoof reaction forces (HRF) were recorded. The carpus was overextended throughout most of the stance. There were 2 periods of overextension, a more rapid period in the beginning of the stance and second directly following the first period. Maximal overextension occurred slightly before the second minimum of the braking horizontal HRF. The metacarpal and antebrachial segments rotated counter-clockwise for most of the stance. The angular velocities of these segments attained absolute and local extremes that were concurrent. The absolute maxima in the longitudinal and the transversal acceleration of the proximal metacarpus were coincident in time with the minimal horizontal braking force. The moment of force about the carpus was extending throughout most of the stance, with a short period of flexion near the end of the stance. The results show that horizontal braking of the hoof results in impulsive loading followed by maximal overextension of the carpus. The second phase of overextension is suggested to be related to the tension or active roll of the flexor tendons of the distal limb. The carpus is, therefore, subjected to rapid and high loading in both the longitudinal and transversal directions, as well as large moments and forces. PMID- 9354298 TI - Variability of the limb joint patterns of sound horses at trot. AB - The reproducibility of gait variables for sound horses is essential for the interpretation of their modifications by locomotor disorders. Therefore, the purpose of this study was to evaluate the variability of the limb joint angle patterns in a population of sound horses while they trotted in the conditions of the routine lameness examination (slow trot, just held by hand, on a track, outdoors). The kinematics of 14 French Saddle horses was recorded using a 3 dimensional (3D) kinematic analysis system. Angle-time diagrams were established in the sagittal plane and their intra- and inter-individual variabilities were evaluated for the whole stride. Horses spontaneously repeated tests at constant speed. The intra- and inter-individual variabilities were low. These values were different from one joint to another and they increased distally. The maximum values were reached by the fetlock and coffin joints. Inter-individual compared to intra-individual variability was 0.9 to 1.9 times greater. There was a high correlation between mean angle-time and mean individual diagrams established for the whole population. It was concluded that it is possible to repeat similar tests with horses trotting in the conditions of the lameness examination. In an homogeneous population, each horse adopts a constant angular pattern that is very close to that of other horses. Since mean angle-time diagrams established on a sound population are highly representative of each mean individual angle pattern, they can be considered as reference data for the further analysis of the pathological gait. PMID- 9354299 TI - Kinematic analysis of the locomotion symmetry of sound horses at a slow trot. AB - This study of the locomotion symmetry was undertaken to provide standard symmetry indices of a group of sound horses at the trot. Using a 3D data collection system, the kinematics of the limb joints of 13 clinically nonlame horses were recorded while trotting in the standard conditions of the clinical lameness examination. A kinematic symmetry indice based on an inter-correlation method was defined and applied to the vertical displacement-time and joint angle-time diagrams of the left and right joints of the horses. For each horse, the mean symmetry indice of each joint was calculated using values from 5 trials. For each joint, these means were then averaged. To evaluate the repeatability of the locomotion symmetry, the intra- and inter-individual variabilities of the symmetry indices were studied. The levels of symmetry of the markers of the trunk were generally lower than those of the limbs. Moreover, during the 5 trials these levels of symmetry were strongly variable but their mean values were very similar from one horse to another. In our experimental conditions the trunk presented a higher degree of freedom than the limbs. This high intra-individual variability indicated also that several trials are necessary to quantify the locomotion symmetry of a trotting horse. In the same way, a lower level of symmetry of the hindlimbs, compared to the forelimbs, was proved by their lower values of symmetry indices. As opposed to the supporting role of the forelimbs, the propulsive role of the hindlimbs may explain this feature. PMID- 9354300 TI - Kinematics in horses at the trot before and after an induced forelimb supporting lameness. AB - The aim of this study was to analyse the biokinematic alterations caused by an induced lameness in the right forelimb of Dutch Warmblood (DWB) horses using a system of computer-aided normal videography. Five mature DWB were recorded with a videocamera (frame rate 1/25) from a lateral view before and after an induced lameness. Before videotaping, passive markers were placed on the skin, over easily identifiable anatomical references to determine the joint angles in the forelimb (always on the flexor side). Lameness was induced using special horseshoes. The lameness was evident at the trot and mild at walk. The images were analysed using a real time digitalising system combined with a previously designed spreadsheet. Linear, temporal and angular parameters (maximum, minimum and angular range of motion) along the stride were calculated as well as the moments of highest extension (Pmax) and flexion (Pmin), expressed as a percentage of the whole stride. Results before and after the induced lameness were compared by a paired Student's t test at a significance level of P<0.05. No differences in speeds before and after the induced lameness were found. Stride length was significantly shorter in the lameness condition. Stride duration was slightly shorter in lameness. The diagonal stance phase increased, while the swing phase decreased. Angular parameters changed mainly in elbow, carpus, fetlock and retraction-protraction angles. This indicated that the angular range of motion in the elbow and carpal joints decreased, and the elbow Pmin occurred later in the stride. The results are useful in the development of video-based equine lameness diagnostics. PMID- 9354301 TI - Compensatory movements of horses with a stance phase lameness. AB - In order to study the mechanism of lameness transfer from fore- and hindlimb lamenesses 2 hypotheses were investigated. Hypothesis 1: Horses with a true supporting limb lameness in one hindlimb show a false supporting limb lameness in the ipsilateral forelimb. Hypothesis 2: Horses with a true supporting limb lameness in one forelimb show a false supporting limb lameness in the contralateral hindlimb. Fourteen horses with fore- or hindlimb lameness were used for this study. Each horse was measured at the trot on a treadmill with standardised speed, before and after diagnostic blocks (9 horses), or with and without induced lameness (5 horses). The head acceleration asymmetry (HAAS) and the sacrum acceleration asymmetry (SAAS) were used for quantification of fore- and hindlimb lameness respectively. Changes were documented by changes of the HAAS or the SAAS. In all 4 horses with a true hindlimb lameness a synchronous false lameness of the ipsilateral forelimb was documented. In 6 of 10 horses with a forelimb lameness a lameness transfer could be assessed according to hypothesis 2. The results of this study show, that horses with a true severe lameness in the forelimb show a false lameness in the contralateral hindlimb, and horses with a true hindlimb lameness show a false lameness in the ipsilateral forelimb. This indicates that the location of the truly lame limb can be deduced from the distribution of 2 lamenesses on a sagittal or diagonal axis. PMID- 9354302 TI - Investigating locomotion of dairy cows by use of high speed cinematography. AB - The longterm influence of management systems on the locomotion of 17 dairy cows was investigated by high speed cinematography (100 frames/s) and kinematic analysis. Angular patterns and hoof trajectories of the left fore- and hindlimbs are presented and statistics made of occurring minimum and maximum angles. At the recording, 3 cows had been kept in tie-stalls (TI) and 6 cows in cubicles (CI) for a consecutive time of about 2.5 years while 8 cows had been kept on grass for about 3 months. Four of the grazing cows had earlier been kept in cubicles (CG) and 4 in tie-stalls (TG) during earlier off grazing seasons together with TI and CI cows. The CI cows had a smaller maximum angle of the elbow joint compared to TI, TG and CG cows. The hock joint angle of the CI cows was less flexed during the stance phase than in TI and CG cows while the minimum angle during the swing phase was greater in the TI and CI cows compared to TG and CG cows. Pastured cows (TG and CG) had a less pronounced flexion of the fetlock joint angle during the stance compared to cows kept indoors (TI and CI). The results suggest that slatted floor and lack of exercise during summer grazing may affect locomotion. This is indicated by restrictions in the movements of the elbow and hock joints and in less fetlock joint flexion at full support. PMID- 9354303 TI - Gait analysis in laying hens and broilers with and without leg disorders. AB - Selection of broiler strains for high body weight has changed the anatomical characteristics and in connection to that, the pattern of locomotion. In addition, rapid growing broilers show a high incidence of leg disorders which compromise the walking ability of the birds. Differences in the patterns of locomotion between laying hens and broilers and between broilers with and without leg disorders can be demonstrated and quantified by gait analysis. The gait pattern was recorded by videotracking. Three points of the body - the cloacal region and both feet joints - were marked by small patches of reflecting foils. The vertical and horizontal movements of the marked points were recorded by a camera in posterior position, while the bird was walking on a treadmill The camera was connected with a PC-operated videotracking system. The vertical and horizontal movements of the 3 marked points were recorded simultaneously and plotted against the time axis. Kinetograms show clearly the differences in the walking pattern between broilers and layers. Layers place the legs directly under the centre of gravity and, therefore, the body moves in a straight line. Broilers, in contrast, move the centre of gravity step by step laterally towards the position of the supporting leg. This pattern may be caused by the anatomical characteristics of broilers. Limping in broilers with leg problems can be measured by differences in the lateral and vertical movements of the right and left leg. PMID- 9354304 TI - Free radicals and antioxidants in cardiovascular disease. AB - Several lines of evidence suggest that free radical-mediated oxidative damage to lipoproteins may be an important factor predisposing them to uptake into the vascular wall. This has led to interest in the factors that determine the susceptibility of lipoproteins to oxidation and their relationship to the development of coronary heart disease. Of these factors, the lipoprotein content of natural antioxidant vitamins such as vitamin E, and beta-carotene have aroused particular interest. Studies in animal models of atherosclerosis suggest that natural and synthetic antioxidants can retard the development of atheroma. Epidemiological comparisons between populations and studies within populations also support the contention that high plasma levels or dietary intake of natural antioxidant vitamins may protect against the development of coronary disease in man. Prospective randomized controlled trials of antioxidants in high risk groups are underway to test whether the theoretical promise of a beneficial role for antioxidants in coronary heart disease prevention will be fufilled. PMID- 9354305 TI - Lipoproteins and cardiovascular reactivity. AB - The observation that relatively short periods of cholesterol lowering therapy can reduce the incidence of coronary artery disease events has prompted interest in the short term effects of lipoproteins on cardiovascular responsiveness. Numerous studies in animals and humans have demonstrated that oxidized LDL-cholesterol can impair endothelial dependent vasodilation in coronary arteries and peripheral resistance vessels. Reduction of plasma LDL-cholesterol levels in hypercholesterolaemic patients improves nitric oxide mediated vasodilator responses in the coronary and peripheral circulation. LDL-cholesterol also potentiates responses to vasoconstrictors such as noradrenaline and endothelin-1 in the absence of endothelium, possibly by enhancing calcium influx into vascular smooth muscle cells. Pharmacological reduction of plasma LDL-cholesterol levels has been shown to reduce blood pressure responses to intravenous infusions of pressor hormones and to stress. However, the relative contribution of changes in endothelial dependent vasodilation and vasoconstrictor or inotropic responses remains to be established. Short term changes in LDL-cholesterol produce changes in cardiovascular responsiveness that may influence the development of ischaemic events. PMID- 9354306 TI - The pharmacokinetics of methadone in healthy subjects and opiate users. AB - AIMS: There is some evidence that monitoring methadone plasma concentration may be of benefit in dosage adjustment during methadone maintenance therapy for heroin (opiate) dependence. However, the kinetics of oral methadone are incompletely characterized. We attempted to describe the latter using a population approach combining intensive 57 h sampling data from healthy subjects with less intensive sparse 24 h data from opiate users. METHODS: Single oral doses of rac-methadone were given to 13 drug-naive healthy subjects (7 men and 6 women) and 17 opiate users beginning methadone maintenance therapy (13 men and 4 women). Plasma methadone concentrations were measured by h.p.l.c. Kinetic analysis was performed using the P-Pharm software. RESULTS: Comparison of kinetic models incorporating mono- or biexponential disposition functions indicated that the latter best represented the data. The improvement was statistically significant for the data from healthy subjects whether the full 57 h or truncated 24 h profiles were used (P<0.031 and P<0.024, respectively), while it was of borderline significance for the more variable data from opiate users (P=0.057) or for pooled (healthy subjects and opiate users) data (P=0.066). The population mean oral clearance of methadone was 6.9+/-1.5 s.d. l h(-1) (5.3+/-1.2 s.d. l h( 1) using 0-24 h data) in the healthy subjects. The results of separate analyses of the data from opiate users and healthy subjects were in contrast with those obtained from pooled data analysis. The former indicated a significantly lower clearance for opiate users (3.2+/-0.3 s.d. l h(-1), P<0.001); 95% CI for the difference = -3 to -6 l h(-1) and no difference in the population mean values of V/F (212+/-27 s.d. l and 239+/-121 s.d. l, P=0.15), while according to the latter analysis addiction was a covariate for V/F but not for oral clearance. A slower absorption of methadone in opiate users was indicated from the analysis of both pooled and separate data. The median elimination half-life of methadone in healthy subjects was 33-46 h depending on the method used to calculate this parameter. CONCLUSIONS: Estimates of the long terminal elimination half-life of methadone (33-46 h in healthy subjects and, possibly, longer in opiate users) indicated that accurate measurement of this parameter requires a duration of sampling longer than that used in this study. Our analysis also suggested that parameters describing plasma concentrations of methadone after a single oral dose in healthy subjects may not be used for predicting and adjusting dosage in opiate users receiving methadone maintenance therapy unless coupled with feedback concentration monitoring techniques (for example Bayesian forecasting). PMID- 9354307 TI - Pharmacokinetics of triptorelin after intravenous bolus administration in healthy males and in males with renal or hepatic insufficiency. AB - AIMS: Triptorelin is a gonadotropin-releasing hormone (GnRH) analogue with enhanced affinity for GnRH receptors and a prolonged half-life due to its resistance to enzymatic degradation. The sustained-release formulation of this molecule is advantageous in conditions requiring chronic hormone suppression. METHODS: This was an open study to determine the pharmacokinetics of a single i.v. bolus dose of 0.5 mg triptorelin acetate in four groups of six male subjects; namely in healthy subjects (Group I), in patients with varying degrees of renal insufficiency (Groups II and III), and in patients with hepatic insufficiency (Group IV). RESULTS: The maximum concentrations of triptorelin were found to be similar for all four study groups (geometric mean Cmax between 41.6 mg ml(-1) and 53.9 mg ml(-1)). The total clearance of triptorelin decreased with increasing renal impairment, and was even lower in patients with hepatic insufficiency (geometric mean CLtot: 210 ml min(-1), 113 ml min(-1), 86.8 ml min( 1) and 57.3 ml min(-1) for Groups I, II, III and IV, respectively). Serum triptorelin concentrations in all four groups were adequately described by a three-compartment model. The elimination half-life for patients with hepatic impairment was similar to that of patients with renal impairment (geometric mean t(1/2, z): 6.6 h, 7.7 h and 7.6 h for Groups II, III and IV, respectively), but significantly longer than in healthy volunteers (2.8 h for Group I). The first and second distribution half-lives were similar for the four groups studied, with geometric mean distribution half-lives of about 0.1 h (6 min) and 0.75 h (45 min), respectively. CONCLUSIONS: Although both renal and hepatic function are important for the clearance of triptorelin, the liver plays the predominant role in subjects suffering from some degree of renal impairment. PMID- 9354308 TI - Decreased oral availability of cyclosporin A at second administration in humans. AB - AIMS: The objective of this study was to determine the extent of period effect on the pharmacokinetics of cyclosporin A (CsA) during consecutive dosing. METHODS: Sandimmune Neoral and Neoplanta capsules were administered to twenty-four healthy Korean male subjects at a single CsA dose of 175 mg in a 2 x 2 crossover investigation with a 2-week wash-out phase. Concentrations of CsA in blood were measured by a r.i.a. method for a period of 48 h. RESULTS: The two formulations were found bioequivalent, but analysis of variance (ANOVA) indicated that there is a significant (P < 0.01) period effect in AUC(0,last) (area under the blood concentration-time curve above the assay limit) and Cmax (maximum blood concentration) between the administrations. A 6 and 9% decrease in the AUC(0,last) and Cmax, respectively was seen at the second administration. CONCLUSIONS: This period effect on the pharmacokinetics of CsA may be relevant for the patients who need consecutive administration of the drug. PMID- 9354309 TI - Relative dispersion of intravascular transit times during isolated human limb perfusions for recurrent melanoma. AB - AIMS: We have characterized the relative dispersion of vascular and extravascular markers in the limbs of three patients undergoing isolated limb perfusions with the cytotoxic melphalan for recurrent malignant melanoma both before and after melphalan dosing. METHODS: A bolus of injectate containing [51Cr] labelled red blood cells, [14C]-sucrose and [3H]-water was injected into an iliac or femoral artery and outflow samples collected at 1 s intervals by a fraction collector. The radioactivity due to each isotype was analysed by either gamma [51Cr] or beta [14C and 3H] counting. The moments of the outflow fraction-time profiles were estimated by a nonparametric (numerical integration) method and a parametric model (sum of two inverse Gaussian functions). RESULTS: The availability, mean transit time and normalised variance (CV2) obtained for labelled red blood cells, sucrose and water were similar before and after melphalan dosing and with the two methods of calculation but varied between the patients. CONCLUSIONS: The vascular space is not well-stirred but characterized by a CV2 similar that reported previously for in situ rat hind limb and rat liver perfusions. A flow-limited blood-tissue exchange was observed for the permeating indicators. Administration of melphalan did not influence the distribution characteristics of the indicators. PMID- 9354310 TI - Inhibition of vasoconstriction by potassium channel opener aprikalim in human conduit arteries used as bypass grafts. AB - AIMS: Potassium channel openers (KCOs) are of potential therapeutic value. Little is known about the effect of these drugs on human conduit arteries used as coronary bypass grafts. The purpose of this study was to determine the effect of the KCO aprikalim (RP52891) on human arteries used as coronary bypass grafts with emphasis on the possible difference in the inhibitory effect on depolarizing agent-mediated rather than receptor-mediated contraction. METHODS: Human internal mammary artery segments (IMA, n = 88) taken from 28 patients were studied. Concentration-relaxation curves for aprikalim were established in IMA precontracted with three vasoconstrictors (K+, U46619, and phenylephrine). In IMA rings incubated with aprikalim (1 or 30 microM) for 10 min concentration contraction curves for the three vasoconstrictors were constructed. RESULTS: Aprikalim-induced relaxation was less in K+ (37.3 +/- 6.4%) than in U46619 (80.2 +/- 7.7%, P=0.002), or phenylephrine (67.5 +/- 7.0%, P=0.038) -precontracted IMA. The EC50 for K+-(-5.40 +/- 0.12 log M) was significantly higher than that for phenylephrine (-6.43 +/- 0.30 log M, P=0.007) but not significant compared with that for U46619 (-5.81 +/- 0.11, P>0.05). Pretreatment with aprikalim depressed the contraction by phenylephrine from 140.6 +/- 27.6% to 49.3 +/- 14.1% (P=0.002) and shifted the EC50 11.0-fold higher in rings treated with 1 microM aprikalim (P=0.007). Treatment of aprikalim did not significantly reduce the K+ and U46619 induced contraction (P>0.05) but shifted the concentration-contraction curves rightward (2.8-fold higher for K+, P<0.05 and 2.2-fold higher for U46619, P<0.05). CONCLUSIONS: This study demonstrates that aprikalim has vasorelaxant effects in human conduit arteries used as coronary artery bypass grafts contracted by a variety of vasoconstrictors and this effect is vasoconstrictor selective with greater potency for alpha1-adrenoceptor agonists than for depolarizing agent K+. These findings provide information on the possible use of this KCO in various clinical settings. PMID- 9354311 TI - Comparison of nitroprusside and nitroglycerin in inhibition of angiotensin II and other vasoconstrictor-mediated contraction in human coronary bypass conduits. AB - AIMS: To compare the effect of nitroprusside (SNP) and nitroglycerin (NTG) on angiotensin II (ANGII), endothelin-1 (ET-1), and alpha1-adrenoceptor (phenylephrine, PE)-mediated contraction in internal mammary artery (IMA). METHODS: Human IMA segments (n=120) taken from 37 patients were studied. Concentration-relaxation curves for SNP and NTG were established in IMA precontracted with these vasoconstrictors. Concentration-contraction curves were also constructed in IMA rings incubated with SNP and NTG (0.1 and 1 microM) for 10 min. RESULTS: Both SNP and NTG caused full relaxation with similar EC50s except NTG was four-fold more potent than SNP in PE-induced contraction (-7.92 +/ 0.06 vs -7.32 +/- 0.2 log M, mean +/- s.e. mean, P<0.01; 95% confidence interval for the difference of the means: 0.19, 1.01 log M). Pretreatment with SNP (0.1 and 1 microM) significantly depressed the contraction by ANGII from 56.6 +/- 7.7% (of 100 mM K+-contraction) to 18.3 +/- 8.6% and 3.9 +/- 2.1% (P=0.0001). In four rings treated with SNP, the contraction to ANGII was abolished whereas NTG did not depress ANGII-mediated contraction. Pretreatment with SNP (1 microM), but not NTG, significantly depressed the magnitude of the PE-induced contraction from 4.7 +/- 1.2 to 1.7 +/- 0.4 g (P<0.05). Treatment with both SNP and NTG significantly increased the EC50 (-5.09 +/- 0.17 log M, P=0.0007 for SNP and -5.40 +/- 0.06 log M, P=0.02 for NTG). Pretreatment with SNP did not significantly change either the magnitude or the EC50 of the ET-1-induced contraction. CONCLUSIONS: SNP may be advantageous compared with NTG in preventing coronary arterial graft contraction. However, once grafts have constricted to ANGII, alpha1-adrenoceptor agonists, and ET-1, NTG may be only marginally advantageous. PMID- 9354312 TI - Effects of losartan on cerebral and ocular circulation in healthy subjects. AB - AIMS: The introduction of specific inhibitors of AT1 receptors, such as losartan, has enabled the investigation of the renin-angiotensin-system (RAS) in humans in vivo. We studied the role of the RAS in the cerebral and ocular circulation in healthy subjects. Haemodynamic effects of orally administered losartan were investigated with non-invasive methods. METHODS: In a placebo-controlled randomized, double-blind two way crossover design losartan (100 mg orally) or placebo was administered in 10 healthy subjects. The effect of losartan was studied at hourly intervals for 8 h. In addition, the effect of losartan on haemodynamic changes induced by exogenous angiotensin II (Ang II) was assessed. Blood flow velocities in the ophthalmic and the middle cerebral artery (OA, MCA) were measured with Doppler sonography. Pulsatile choroidal blood flow was estimated with laser interferometric measurement of fundus pulsation. RESULTS: Losartan significantly increased fundus pulsation amplitude (+11%, 95% CI: 5 to 16% P<0.0001), tended to increase resistive index in the ophthalmic artery (+12%, 95% CI: 0 to 23%) and tended to decrease mean arterial pressure (-15%, 95% CI: 23 to -1%). Ang II induced effects on cerebral, ocular and systemic haemodynamics were prevented by preceding administration of losartan. CONCLUSIONS: The present data suggest that Ang II is not a major determinant of cerebral and ocular blood flow in vivo. The observed changes in cerebral and ocular haemodynamic parameters after losartan administration reflect effects on systemic blood pressure. PMID- 9354314 TI - Influence of SNN-6010, an elemental diet, on the generation of cytokines, NO, and chemiluminescence from leukocytes and umbilical vein endothelial cells in man. AB - AIMS: There is insufficient information on the influence of elemental diet (ED) on leukocytes and endothelial cells although ED has been used increasingly for debilitated and post-operative patients. We here aimed to show the influence of SNN-6010 (Twinline; TL), which contains medium-chain triglycerides, on the generation of cytokines and nitric oxide (NO) and chemiluminescence in human umbilical vein endothelial cells (HUVEC) and leukocytes. METHODS: Leukocytes were obtained from 10 healthy volunteers and 10 patients with jaw fractures who underwent TL for 30 days after intermaxillary fixation. Luminol-dependent chemiluminescence and NO generation were measured in polymorphonuclear leukocytes (PMN) and HUVEC and generation of interleukin-1 beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha), and prostaglandin E2 (PGE2) as well as expression of these cytokines' mRNA were examined in lymphocytes, monocytes, PMN, and HUVEC. RESULTS: TL dose-dependently suppressed generation of IL-1beta, TNF-alpha, PGE2, and NO in all kinds of cells from healthy individuals. Lymphocytes from patients who underwent TL feeding for 30 days showed reduced generation of IL-1beta and TNF-alpha, and the suppressed cytokine generation corresponded with the decreased expression of mRNA of these cytokines. Chemiluminescence by PMN from healthy individuals was suppressed by TL dose-dependently, and PMN from TL-fed patients showed reduced chemiluminescence. CONCLUSION: These results appear to indicate that TL suppresses the functions of leukocytes and endothelial cells by down regulating the generation of reactive oxygen intermediates, NO, and cytokines. PMID- 9354313 TI - The peptide endothelin receptor antagonist, TAK-044, produces sustained inhibition of endothelin-1 mediated arteriolar vasoconstriction. AB - AIMS: Endothelin-1 (ET-1) has been implicated in the pathophysiology of a number of cardiovascular diseases for which endothelin receptor antagonists are currently under clinical development. We have previously reported that systemic administration of the combined endothelin A/B receptor antagonist, TAK-044, abolishes the forearm vasoconstriction caused by intrabrachial ET-1 infusion for at least 3 h. In this study we investigated whether TAK-044 can inhibit ET-1 mediated forearm vasoconstriction for longer periods. METHODS: Eighteen subjects were recruited to a randomized, placebo-controlled, single-blind, three-way, crossover study. Subjects were divided into three groups of six. Groups received 25 mg, 50 mg or 100 mg TAK-044 on two separate occasions, 6 and 10 h before the start of a 2 h intrabrachial infusion of ET-1 (5 pmol min(-1)). On a third occasion subjects received only placebo before intra-arterial ET-1 infusion. Forearm vasoconstriction to ET-1 was measured by venous occlusion plethysmography. RESULTS: In the placebo phase, ET-1 caused significant, slowly progressive local forearm vasoconstriction of approximately 30% (P<0.01) in all three groups. All three doses of TAK-044, administered at both timepoints, tended to blunt the vasoconstriction caused by ET-1. When the responses from all three groups were combined, TAK-044 significantly reduced ET-1 mediated vasoconstriction compared with placebo -9% (95% CI -15 to -3; P=0.01) at 8 h and by -9% (95% CI -17 to -2; P=0.01) 12 h after dosing. CONCLUSIONS: TAK-044 attenuated, but did not abolish, local ET-1 mediated vasoconstriction, for up to 12 h after administration. Vasoconstriction to local intra-arterial administration of ET-1 appears to represent a safe and reproducible pharmacodynamic index of systemic endothelin receptor antagonism in humans. PMID- 9354315 TI - An evaluation of the interaction of meloxicam with frusemide in patients with compensated chronic cardiac failure. AB - AIMS: To evaluate the interaction of meloxicam with frusemide in patients with compensated cardiac failure. METHODS: Nineteen patients with Grade II or III compensated chronic cardiac failure completed this randomized, double-blind, cross-over study. The patients received 40 mg frusemide day(-1) for 7 days. Thereafter, patients received either 15 mg meloxicam plus 40 mg frusemide day( 1), or one placebo tablet plus 40 mg frusemide day(-1) for 7 days. After a washout period of 7 days during which patients received 40 mg frusemide day(-1) for 7 days, the patients were crossed over to the alternate treatment. The effect of concomitant ingestion of meloxicam and frusemide on frusemide-induced diuresis, urine and serum electrolytes, urinary frusemide excretion, and plasma frusemide pharmacokinetics was also determined. RESULTS: The estimate (90% confidence interval) of the '(frusemide + meloxicam)/(frusemide alone)' mean ratio of the variables Cmax, AUC(SS) and Cmax/AUC(SS) for plasma frusemide were 121% (101% to 145%), 106% (96.4% to 117%), and 114% (98.3% to 132%), respectively. Similarly, the estimate (90% confidence interval) of the '(frusemide + meloxicam)/(frusemide alone)' of the mean ratio of the variable cumulative urinary frusemide excretion after multiple doses of frusemide were 123% (101% to 150%) for the period 0-8 h, and 122% (105% to 142%) for the period 0-24 h after drug administration on day 7. The estimate (90% confidence interval) of the '(frusemide + meloxicam)/(frusemide alone)' mean ratio of the pharmacodynamic variables cumulative sodium excretion was 105% (95.2% to 116%) for the period 0-8 h and 108% (96.5% to 121%) for the period 0-24 h after drug administration on day 7. CONCLUSIONS: Meloxicam may lead to slightly increased maximum concentrations of frusemide in plasma, as well as to slightly increased urinary excretion of frusemide, without affecting the pharmacodynamics of frusemide. Thus there is no clinically significant pharmacokinetic or pharmacodynamic interaction of meloxicam with frusemide following repeated co administration of meloxicam and frusemide to patients with compensated chronic cardiac failure. PMID- 9354316 TI - No effect of short-term omeprazole intake on acenocoumarol pharmacokinetics and pharmacodynamics. AB - AIMS: To investigate the effect of omeprazole on the pharmacokinetics of R- and S acenocoumarol and on their combined anticoagulant activity. METHODS: Eight healthy male subjects completed a double-blind, randomized, placebo-controlled, two-way cross-over study. Subjects were given either omeprazole 40 mg or placebo once daily for 3 days. On day 2 of each study period, a single 10 mg oral dose of racemic acenocoumarol was administered and venous blood samples were collected for pharmacokinetic and pharmacodynamic assessments. A wash-out period of 2 weeks separated the two study periods. RESULTS: The pharmacokinetics of R- and S acenocoumarol (AUC 3016 +/- 221 and 233 +/- 14 ng ml(-1) h, respectively) did not change after omeprazole (AUC 2929 +/- 256 and 220 +/- 18 ng ml(-1) h, respectively). Anticoagulant activity (INRmax 1.7 +/- 0.1) was unaffected by co administration of omeprazole (INRmax 1.7 +/- 0.1). CONCLUSIONS: The short-term intake of omeprazole does not affect acenocoumarol pharmacokinetics or pharmacodynamics. These data differ from the results of previous studies on the effect of omeprazole on warfarin, suggesting a different in vivo interaction profile of omeprazole on acenocoumarol than on warfarin. Drug interaction studies with oral anticoagulants should not be restricted to the use of warfarin. PMID- 9354317 TI - Effects of tedisamil, atenolol and their combination on heart and rate-dependent QT interval in healthy volunteers. AB - AIMS: Tedisamil is a new blocker of K+ currents in cardiac tissues, exerts bradycardic effects and has shown clinical efficacy in angina pectoris. Theoretically, when coadministered with a beta-adrenoceptor blocker the tedisamil combination could induce dangerous bradycardia and QT interval prolongation. Therefore, the aim of this study was to evaluate the effects of tedisamil and atenolol alone and in combination, on heart rate and QT interval duration at rest and during exercise tests. METHODS: The effects of tedisamil (100 mg twice daily) and atenolol (50 mg twice daily) on heart rate and QT interval duration were analysed in a three-period crossover study in healthy male volunteers. RESULTS: This study showed that tedisamil exerted a significant (P<0.05) bradycardic action at rest (-10 beats min(-1); 95% CI: -6 to -15 beats min(-1)) similar to atenolol (-14 beats min(-1); -11 to -17) and drug combination (-9 beats min(-1); 6 to -12). During exercise, at the highest comparable workload, heart rate did not decrease significantly with tedisamil but decreased significantly with atenolol (-42 beats min(-1); -37 to -47) and combination (-47 beats min(-1); -41 to 52). Atenolol did not modify QT interval duration. Tedisamil alone and in combination with atenolol increased QT interval duration by 12% (95% CI: 7 to 17%) and 12% (8 to 16) respectively at RR=1000 ms, but not at RR<700 ms (combination). Tedisamil alone and in combination induced a reverse rate dependent QT interval prolongation. CONCLUSIONS: These results indicate that the combination of tedisamil and atenolol is not associated with excessive bradycardia or excessive QT interval prolongation in healthy subjects. PMID- 9354318 TI - Brodimoprim: adverse drug reactions from spontaneous reporting. PMID- 9354319 TI - Aminophylline and essential tremor. PMID- 9354320 TI - A genetically encoded optical probe of membrane voltage. AB - Measuring electrical activity in large numbers of cells with high spatial and temporal resolution is a fundamental problem for the study of neural development and information processing. To address this problem, we have constructed a novel, genetically encoded probe that can be used to measure transmembrane voltage in single cells. We fused a modified green fluorescent protein (GFP) into a voltage sensitive K+ channel so that voltage-dependent rearrangements in the K+ channel would induce changes in the fluorescence of GFP. The probe has a maximal fractional fluorescence change of 5.1%, making it comparable to some of the best organic voltage-sensitive dyes. Moreover, the fluorescent signal is expanded in time in a way that makes the signal 30-fold easier to detect. A voltage sensor encoded into DNA has the advantage that it may be introduced into an organism noninvasively and targeted to specific developmental stages, brain regions, cell types, and subcellular compartments. PMID- 9354321 TI - A novel in vitro preparation: the intact hippocampal formation. AB - The intact hippocampal formation (IHF) of neonatal or young rats can be kept alive for an extended period in a fully submerged chamber with excellent morphological preservation. Field or patch-clamp recordings, intracellular Ca2+ measurements, and 3-D reconstruction of biocytin-filled neurons can be performed routinely. The generation and propagation of network-driven activities can be studied within the IHF or between connected intact structures such as the septum and the hippocampus or two hippocampi, and the use of a dual chamber enables the application of drugs separately to each structure. This preparation will be useful to study intact neuronal networks in the developing hippocampus in vitro. PMID- 9354322 TI - Better late than never: a role for rabs late in exocytosis. PMID- 9354323 TI - Mutant mice and neuroscience: recommendations concerning genetic background. Banbury Conference on genetic background in mice. PMID- 9354324 TI - Specification of mouse telencephalic and mid-hindbrain progenitors following heterotopic ultrasound-guided embryonic transplantation. AB - We have demonstrated the utility of ultrasound backscatter microscopy for targeted intraparenchymal injections into embryonic day (E) 13.5 mouse embryos. This system has been used to test the degree of commitment present in neural progenitors from the embryonic ventral telencephalon and mid-hindbrain region. Many E13.5 ventral telencephalic progenitors were observed to integrate and adopt local phenotypes following heterotopic transplantation into telencephalic or mid hindbrain targets, whereas mid-hindbrain cells of the same stage were unable to integrate and change fate in the telencephalon. In contrast, many mid-hindbrain cells from an earlier developmental stage (E10.5) were capable of integrating and adopting a forebrain phenotype after grafting into the telencephalon, suggesting that mouse mid-hindbrain progenitors become restricted in their developmental potential between E10.5 and E13.5. PMID- 9354325 TI - Isolation of lineage-restricted neuronal precursors from multipotent neuroepithelial stem cells. AB - We have identified a neuronal-restricted precursor (NRP) cell that expresses E NCAM (high polysialic-acid NCAM) and is morphologically distinct from multipotent neuroepithelial (NEP) cells (Kalyani et al., 1997) and spinal glial progenitors (Rao and Mayer-Proschel, 1997). NRP cells self renew over multiple passages in the presence of fibroblast growth factor (FGF) and neurotrophin-3 (NT-3) and differentiate in the presence of retinoic acid and the absence of FGF into postmitotic neurons. NRP cells can also be generated from multipotent E10.5 NEP cells. Clonal analysis shows that NRP cells arise from a NEP progenitor that generates other restricted CNS precursors. The NEP-derived NRPs undergo self renewal and can differentiate into multiple neuronal phenotypes. Thus, a direct lineal relationship exists between multipotential NEP cells and more restricted neuronal precursor cells present in vivo at E13.5 in the spinal cord. PMID- 9354327 TI - Activity regulates the synaptic localization of the NMDA receptor in hippocampal neurons. AB - We describe here a novel effect of activity on the subcellular distribution of NMDA receptors in hippocampal neurons in culture. In spontaneously active neurons, NMDA receptors were clustered at a few synaptic and nonsynaptic sites. Chronic blockade of NMDA receptor activity induced a 380% increase in the number of NMDA receptor clusters and a shift to a more synaptic distribution. This effect was reversible. The distributions of the presynaptic marker synaptophysin, the AMPA-type glutamate receptor subunit GluR1, and the putative NMDA receptor clustering protein PSD-95 were not affected by blockade. Regulation of the synaptic localization of NMDA receptors by activity may define a novel mechanism by which input controls a neuron's ability to modify its synapses. PMID- 9354326 TI - Synaptic clustering of the cell adhesion molecule fasciclin II by discs-large and its role in the regulation of presynaptic structure. AB - The cell adhesion molecule Fasciclin II (FASII) is involved in synapse development and plasticity. Here we provide genetic and biochemical evidence that proper localization of FASII at type I glutamatergic synapses of the Drosophila neuromuscular junction is mediated by binding between the intracellular tSXV bearing C-terminal tail of FASII and the PDZ1-2 domains of Discs-Large (DLG). Moreover, mutations in fasII and/or dlg have similar effects on presynaptic ultrastructure, suggesting their functional involvement in a common developmental pathway. DLG can directly mediate a biochemical complex and a macroscopic cluster of FASII and Shaker K+ channels in heterologous cells. These results indicate a central role for DLG in the structural organization and downstream signaling mechanisms of cell adhesion molecules and ion channels at synapses. PMID- 9354328 TI - Disruption of a behavioral sequence by targeted death of peptidergic neurons in Drosophila. AB - The neuropeptide eclosion hormone (EH) is a key regulator of insect ecdysis. We tested the role of the two EH-producing neurons in Drosophila by using an EH cell specific enhancer to activate cell death genes reaper and head involution defective to ablate the EH cells. In the EH cell knockout flies, larval and adult ecdyses were disrupted, yet a third of the knockouts emerged as adults, demonstrating that EH has a significant but nonessential role in ecdysis. The EH cell knockouts had discrete behavioral deficits, including slow, uncoordinated eclosion and an insensitivity to ecdysis-triggering hormone. The knockouts lacked the lights-on eclosion response despite having a normal circadian eclosion rhythm. This study represents a novel approach to the dissection of neuropeptide regulation of a complex behavioral program. PMID- 9354329 TI - Potentiation of developing synapses by postsynaptic release of neurotrophin-4. AB - The hypothesis that synaptic functions can be regulated by neurotrophins secreted from the postsynaptic cell was examined in Xenopus nerve-muscle cultures. Neuromuscular synapses formed on myocytes overexpressing neurotrophin-4 (M+ synapses) exhibited a higher level of spontaneous synaptic activity and enhanced evoked synaptic transmission as compared to those formed on normal control myocytes (M- synapses). The NT-4 effects involve a potentiation of presynaptic transmitter secretion as well as a lengthening of the mean burst duration of postsynaptic low conductance acetylcholine channels. Repetitive stimulation of either the presynaptic neuron or the postsynaptic myocyte led to a potentiation of synaptic transmission at M+ synapses. All potentiation effects of NT-4 overexpression were abolished by the extracellular presence of TrkB-IgG but not by the presence of TrkA-IgG, indicating that postsynaptic secretion of NT-4 was responsible for the synaptic modification. PMID- 9354330 TI - Dopamine D3 receptor mutant mice exhibit increased behavioral sensitivity to concurrent stimulation of D1 and D2 receptors. AB - The dopamine D3 receptor is expressed primarily in regions of the brain that are thought to influence motivation and motor functions. To specify in vivo D3 receptor function, we generated mutant mice lacking this receptor. Our analysis indicates that in a novel environment, D3 mutant mice are transiently more active than wild-type mice, an effect not associated with anxiety state. Moreover, D3 mutant mice exhibit enhanced behavioral sensitivity to combined injections of D1 and D2 class receptor agonists, cocaine and amphetamine. However, the combined electrophysiological effects of the same D1 and D2 agonists on single neurons within the nucleus accumbens were not altered by the D3 receptor mutation. We conclude that one function of the D3 receptor is to modulate behaviors by inhibiting the cooperative effects of postsynaptic D1 and other D2 class receptors at systems level. PMID- 9354332 TI - Brain regions differentially involved in remembering what and when: a PET study. AB - Recollecting a past episode involves remembering not only what happened but also when it happened. We used positron emission tomography (PET) to directly contrast the neural correlates of item and temporalorder memory. Subjects studied a list of words and were then scanned while retrieving information about what words were in the list or when they occurred within the list. Item retrieval was related to increased neural activity in medial temporal and basal forebrain regions, whereas temporal-order retrieval was associated with activations in dorsal prefrontal, cuneus/precuneus, and right posterior parietal regions. The dissociation between temporal and frontal lobe regions confirms and extends previous lesion data. The results show that temporal-order retrieval involves a network of frontal and posterior brain regions. PMID- 9354331 TI - IB4-binding DRG neurons switch from NGF to GDNF dependence in early postnatal life. AB - We have tested the role of glial cell line-derived neurotrophic factor (GDNF) in regulating a group of putatively nociceptive dorsal root ganglion (DRG) neurons that do not express calcitonin gene-related peptide (CGRP) and that downregulate the nerve growth factor (NGF) receptor tyrosine kinase, TrkA, after birth. We show that mRNA and protein for the GDNF receptor tyrosine kinase, Ret, are expressed in the DRG in patterns that differ markedly from those of any of the neurotrophin receptors. Most strikingly, a population of small neurons initiates expression of Ret between embryonic day 15.5 and postnatal day 7.5 and maintains Ret expression into adulthood. These Ret-expressing small neurons are selectively labeled by the lectin IB4 and project to lamina IIi of the dorsal horn. Ret expressing neurons also express the glycosyl-phosphatidyl inositol-linked (GPI linked) GDNF binding component GDNFR-alpha and retrogradely transport 125I-GDNF, indicating the presence of a biologically active GDNF receptor complex. In vitro, GDNF supports the survival of small neurons that express Ret and bind IB4 while failing to support the survival of neurons expressing TrkA and CGRP. Together, our findings suggest that IB4-binding neurons switch from dependence on NGF in embryonic life to dependence on GDNF in postnatal life and are likely regulated by GDNF in maturity. PMID- 9354333 TI - Relationships between local synaptic connections and orientation domains in primary visual cortex. AB - Combined optical imaging of ferret primary visual cortex in vivo and scanning laser photostimulation in brain slices were used to determine the spatial relationships between synaptic inputs onto individual neurons and the pattern of orientation columns. In the upper cortical layers, both excitatory and inhibitory inputs originated primarily from regions with orientation tuning similar to that of the recorded neurons; the shapes of the input tuning curves were indistinguishable. The orientation distributions of both types of inputs centered around the orientation of the recorded neurons, and no evidence for preferential cross-orientation inputs, either excitatory or inhibitory, was observed. These patterns of synaptic connectivity are most consistent with feedforward models for generation of orientation selectivity and are inconsistent with the patterns required by models based on cross-orientation inhibition. PMID- 9354334 TI - Altered subthreshold sodium currents and disrupted firing patterns in Purkinje neurons of Scn8a mutant mice. AB - Sodium currents and action potentials were characterized in Purkinje neurons from ataxic mice lacking expression of the sodium channel Scn8a. Peak transient sodium current was approximately 60% of that in normal mice, but subthreshold sodium current was affected much more. Steady-state current elicited by voltage ramps was reduced to approximately 30%, and resurgent sodium current, an unusual transient current elicited on repolarization following strong depolarizations, was reduced to 8%-18%. In jolting mice, with a missense mutation in Scn8a, steady state and resurgent current were also reduced, with altered voltage dependence and kinetics. Both spontaneous firing and evoked bursts of spikes were diminished in cells from null and jolting mice. Evidently Scn8a channels carry most subthreshold sodium current and are crucial for repetitive firing. PMID- 9354335 TI - Kainate receptors presynaptically downregulate GABAergic inhibition in the rat hippocampus. AB - Using microcultured neurons and hippocampal slices, we found that under conditions that completely block AMPA receptors, kainate induces a reduction in the effectiveness of GABAergic synaptic inhibition. Evoked inhibitory postsynaptic currents (IPSCs) were decreased by kainate by up to 90%, showing a bell-shaped dose-response curve similar to that of native kainate-selective receptors. The down-regulation of GABAergic inhibition was not affected by antagonism of metabotropic receptors, while it was attenuated by CNQX. Kainate increased synaptic failures and reduced the frequency of miniature IPSCs, indicating a presynaptic locus of action. In vivo experiments using brain dialysis demonstrated that kainate reversibly abolished recurrent inhibition and induced an epileptic-like electroencephalogram (EEG) activity. These results indicate that kainate receptor activation down-regulates GABAergic inhibition by modulating the reliability of GABA synapses. PMID- 9354336 TI - Dendritically released peptides act as retrograde modulators of afferent excitation in the supraoptic nucleus in vitro. AB - Oxytocin (OXT) and vasopressin (VP) are known to be released from dendrites of magnocellular neurons. Here, we show that these peptides reduced evoked EPSCs by a presynaptic mechanism, an effect blocked by peptide antagonists and mimicked by inhibition of endogenous peptidases. Dendritic release of peptides, elicited with depolarization achieved by high frequency stimulation of afferents or with current injection into an individual neuron, induced short-term synaptic depression similar to that seen following exogenous peptide application and was prevented by peptide antagonists. Thus, dendritically released peptides depress evoked EPSCs in magnocellular neurons by activating presynaptic OXT and/or VP receptors. Such a retrograde modulatory action on afferent excitation may serve as a feedback mechanism to permit peptidergic neurosecretory neurons to autoregulate their own activity. PMID- 9354337 TI - Identification of amino acid residues that control functional behavior in GluR5 and GluR6 kainate receptors. AB - GluR5 and GluR6 kainate receptors differ in their responses to a variety of agonists, despite their relatively high primary sequence homology. We carried out a structure-function study to identify amino acids underlying these divergent responses. Patch clamp analysis of chimeric GluR5-GluR6 receptors indicated that several functionally dominant sites were localized to the C-terminal side of M1. All nonconserved amino acids in the region between M3 and M4 of GluR6 were then individually mutated to their GluR5 counterparts. We found that a single amino acid (N721 in GluR6) controls both AMPA sensitivity and domoate deactivation rates. Additionally, mutation of A689 in GluR6 slowed kainate desensitization. These functional effects were accompanied by alterations in binding affinities. These results support a critical role for these residues in receptor binding and gating activity. PMID- 9354338 TI - Changes in permeability caused by connexin 32 mutations underlie X-linked Charcot Marie-Tooth disease. AB - The relationship between the loss of connexin 32 function and clinical manifestations of X-linked Charcot-Marie-Tooth (CMTX) disease is unknown. Here, we report that eight of nine CMTX mutations investigated form channels with measurable electrical conductance. Single-channel studies of two mutations demonstrate reduced junctional permeability caused by a decrease in either pore size (S26L) or open channel probability (M34T) that favors residency in a low conductance substate. Permeation of second messengers such as cAMP through reflexive gap junctions between adjacent cytoplasmic loops of myelinating Schwann cells is likely to be reduced or absent in these channels. We propose that CMTX mutations impair the transduction of signals arising from normal glial-neuronal interactions and thereby cause demyelination and axonal degeneration. PMID- 9354339 TI - Accelerated amyloid deposition in the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid precursor proteins. AB - Missense mutations in two related genes, termed presenilin 1 (PS1) and presenilin 2 (PS2), cause dementia in a subset of early-onset familial Alzheimer's disease (FAD) pedigrees. In a variety of experimental in vitro and in vivo settings, FAD linked presenilin variants influence the processing of the amyloid precursor protein (APP), leading to elevated levels of the highly fibrillogenic Abeta1-42 peptides that are preferentially deposited in the brains of Alzheimer Disease (AD) patients. In this report, we demonstrate that transgenic animals that coexpress a FAD-linked human PS1 variant (A246E) and a chimeric mouse/human APP harboring mutations linked to Swedish FAD kindreds (APP swe) develop numerous amyloid deposits much earlier than age-matched mice expressing APP swe and wild type Hu PS1 or APP swe alone. These results provide evidence for the view that one pathogenic mechanism by which FAD-linked mutant PS1 causes AD is to accelerate the rate of beta-amyloid deposition in brain. PMID- 9354340 TI - Chlamydia pneumoniae in arteries: a tale of the unexpected. PMID- 9354341 TI - Fungal endocarditis. PMID- 9354342 TI - Toxoplasmosis, behaviour and personality. AB - The clinical sequelae of acute and congenital toxoplasmosis are well established, but that of chronic toxoplasma infection remains uncertain. In rodents, chronic toxoplasma infection is associated with altered behaviour leading to an enhanced risk of feline predation and a putative selective advantage to the parasite. It is proposed that neurotropic cysts of toxoplasma exert an effect on animal behaviour, either directly or via the release of metabolic products. Long standing toxoplasma infection in humans has been linked to cerebral tumour formation and personality shift. In view of the vast population with chronic toxoplasma infection, further studies of the clinical sequelae of this condition are required. PMID- 9354343 TI - Primary HIV infection: follow-up of patients initially randomized to zidovudine or placebo. AB - A randomized trial of 77 patients with primary HIV infection concluded that a 6 month course of zidovudine significantly reduced the incidence of opportunistic infections and improved CD4 cell counts. Follow-up was extended over a mean of 28 months to assess whether these benefits persisted beyond the trial period under conditions of routine care. In the post-trial period, antiviral drugs were prescribed with similar frequency in the zidovudine (248 treatment days per patient) and placebo arms (239 days, P=0.59). At the end of follow-up, patients initially assigned to zidovudine had still accumulated fewer disease progression events (three) than those given placebo (eight), but contrary to the initial analysis, the difference was no longer statistically significant (logrank test, P=0.07). The incidence of progression events tended to increase over time in the zidovudine group, and to decrease in the placebo group (trends tested in Weibull survival models, P=0.04). Similarly, initial gains in CD4 counts in the zidovudine arm diminished progressively after the trial, as CD4 counts declined faster in this group than in placebo-treated patients (-5.0 vs. -3.2 cells/mm3/month, P=0.06). In summary, initial clinical and immunological benefits of a 6-month course of zidovudine in patients with primary HIV infection eroded over time, suggesting that longer and more potent antiviral treatment should be given consideration. PMID- 9354345 TI - Routine surveillance blood cultures: their place in the management of critically ill patients. AB - The use of surveillance blood cultures has been advocated as a means to allow earlier detection of septic episodes amongst intensive care patients, and therefore earlier institution of appropriate antibiotic therapy. We compared the results of surveillance cultures and clinically indicated blood cultures for bacterial isolates grown and the influence of culture results on patient management. Blood cultures were obtained from all intensive care unit (ICU) patients over the course of 3 months at a set surveillance time (surveillance group) or according to clinical indications (clinical group). Bacteriological results were compared and real-time chart review performed to assess the influence of the surveillance cultures on patient management, with particular reference to antibiotic therapy. Two hundred and forty-nine blood culture sets were collected over 3 months, 99 in the surveillance group and 150 in the clinical group. A total of 256 bacterial isolates were grown, 95 in the surveillance group and 161 in the clinical group. For the surveillance group 36%, 20%, and 44% of the isolates represented bacteraemia, line colonization and culture contamination, respectively. For the clinical group the distibution was 69%, 7%, and 24% respectively (P<0.001, P<0.01, and P<0.0027 for comparisons of percentages within each classification). On only one occasion was antibiotic therapy started based on the result of a surveillance culture, and on only one occasion was a septic episode detected earlier by a surveillance culture; however, this culture result did not lead to a change in patient management. Surveillance blood cultures are expensive and add very little to the management of patients in the intensive care environment. PMID- 9354344 TI - Evaluation of a score system for the severity and outcome of cytomegalovirus interstitial pneumonia in allogeneic bone marrow recipients. AB - We verified whether a clinical score system developed for renal transplant patients predicts the severity and outcome of cytomegalovirus interstitial pneumonia (CMV IP) in allogeneic bone marrow (BMT) recipients. The score system was retrospectively applied to 20 patients at the estimated date of onset of IP and 10-14 days later. Seven patients received ganciclovir (GCV), seven received GCV plus intravenous immunoglobulin (i.v. Ig), and six received only supportive care. Nine out of 14 patients who received GCV with or without i.v. Ig survived the episode of IP (the median score of these patients at diagnosis of CMV IP was 5 (range 3-8)), while the remaining five patients died of respiratory failure during IP and at the diagnosis had a median score of 10 (range 9-11) (P=0.01). The six patients who received only supportive care survived for a median time of 21 days (range 10-24 days) from the estimated onset of CMV IP, and the median score at the diagnosis of IP was 10 (range 8-12). The overall survival correlates strongly with low initial severity of IP as measured by this score system: 11 out of 20 patients who died of respiratory failure during IP had at the estimated onset of IP a score >8, while of the nine patients who survived IP, eight had at the onset a score <7 and the remaining one a score of 8 (P=0.0007). The sensibility, specificity, predictive positive value and predictive negative value of the score system (with a threshold value of 8) to identify patients who survived IP was: 100%, 88%, 91% and 100%, respectively. The use of ganciclovir alone or in combination was the most important determinant of outcome. These data support the relevance of this score system with a threshold value of 8; if prospective and controlled studies confirm our observations, it would help physicians to identify BMT recipients during CMV IP with high vs. low risk of poor outcome. PMID- 9354346 TI - Investigation of an outbreak of multidrug resistant tuberculosis among renal patients using rpo B gene sequencing and IS6110 inverse PCR. AB - A cluster of cases of tuberculosis among five patients receiving treatment for renal failure was investigated. Insertion sequence (IS6110) fingerprinting and antibiotic resistance profiling of the Mycobacterium tuberculosis isolates from four of the patients (A-D), who had been on the same ward, showed that three of these cases (A-C) were related, but that the fourth (D) was distinct. An isolate from the fifth patient (E), who had been on a separate ward, was indistinguishable from the outbreak strain by IS6110 profile. However, the isolate from patient E and a second isolate from patient A differed from the previous strains in being rifampicin resistant. Sequence analysis of the rpo B genes of the two rifampicin-resistant strains demonstrated the presence of different mutations, showing that they had evolved independently from the same source strain. IS6110 and rpo B gene analyses are invaluable for the accurate investigation of outbreaks of multidrug resistant tuberculosis. PMID- 9354347 TI - Changes in the oral streptococcal flora of children undergoing allogeneic bone marrow transplantation. AB - The changes in the oral streptococcal flora of twenty children undergoing allogeneic bone marrow transplant are described. Saliva was collected from each child on four separate occasions: (i) before the conditioning regimen; (ii) 7 days post-transplantation; (iii) when the neutrophil count had risen above 0.5 x 10(9)/l; (iv) 119 days post-transplantation. Indices for dental caries, plaque, gingivitis, herpetic stomatitis and mucositis were also recorded. There was a significant decrease in the total aerobic (P<0.001) and anaerobic counts (P<0.0002) between baseline and 7 days post-transplantation. The proportion of the 'Streptococcus oralis group' (Streptococcus mitis and S. oralis) increased significantly from baseline 12.1% to 48.4% at 7 days post-transplantation (P<0.003). The plaque and gingivitis indices increased significantly from baseline to 7 days post-transplantation (P<0.001). Twenty percent of the children had either positive blood cultures or Hickman line cultures for the 'S. oralis group', and it is possible that the inflamed gingival tissues are a further site of entry for these streptococci. There were no differences in the total anaerobic counts or the proportion of the 'S. oralis group' between baseline and the end of the study in the transplant children, or between the transplant and control children. PMID- 9354348 TI - N-acetylcysteine treatment and the risk of toxic reactions to trimethoprim sulphamethoxazole in primary Pneumocystis carinii prophylaxis in HIV-infected patients. AB - In a randomized double blind placebo controlled trial, HIV sero-positive patients with CD4+ cell count less than 200 x 10(6)/l or an AIDS diagnosis were evaluated for drug reactions to trimethoprim-sulphamethoxazole (TMP-SMX) during treatment, including pretreatment, with N-acetylcysteine (NAC) 800 mg daily or placebo. TMP SMX (one double-strength tablet containing 160 mg of trimethoprim and 800 mg of sulphamethoxazole) was given three times weekly as primary Pneumocystis carinii (PCP) prophylaxis. Thirty percent (n = 15) of the patients experienced adverse reactions 8-20 (mean 12.7) days after starting with TMP-SMX. At entry, low cysteine and glutathione levels in plasma were found in the HIV-positive patients. Age, sex, CD4+ count, plasma cysteine and glutathione levels were not risk factors for adverse reactions to TMP-SMX. However, concomitant therapy with nucleoside analogues was associated with increased risk for TMP-SMX reactions. Oral NAC 800 mg daily was well tolerated, but replenished neither cysteine nor glutathione levels in plasma. NAC 800 mg/day did not significantly decrease the risk of adverse reactions to TMP-SMX in this study, and could thus not be recommended for this purpose. A prolonged pretreatment period and/or higher dose of NAC may be necessary for clinical effect. PMID- 9354349 TI - Evaluation of immunohistochemistry for the detection of Helicobacter pylori in gastric mucosal biopsies. AB - A reliable diagnosis of Helicobacter pylori is important in clinical practice and research. To evaluate sensitivity, specificity and inter-observer variation of three histological staining methods for the diagnosis of intragastric H. pylori bacterial flora, four observers assessed the presence of H. pylori in a test set of pairs of gastric biopsies taken from 40 patients during GI-endoscopy. Histological slides of one biopsy of each patient (formalin-fixed and paraffin embedded) were stained with a Modified Giemsa (MG), the Warthin-Starry (WS), and an immunohistochemical method (IMM) using purified polyclonal H. pylori antiserum (DAKO B471). As a standard, bacterial culture was performed on the adjacent biopsy, using the four quadrants method. Special attention was paid to the presence of non-H. pylori bacterial flora. Twenty out of 40 specimens were H. pylori-positive by culture. Using culture as a standard, sensitivity for H. pylori was 90.0+/-10.0% with MG, 70.0+/-14.1% with WS, and 83.8+/-11.1% with IMM stain. Specificity was 53.8+/-19.3%, 82.5+/-9.6% and 90.0+/-0.0%, respectively. Of 37, 14 and eight false positive scores by all observers for MG, WS and IMM, respectively, non-H. pylori flora was cultured from 17, six, and four of the corresponding adjacent biopsies. Non-H. pylori flora may account for overdiagnosis of H. pylori in gastric biopsies. Kappa values for the variance between the four observers were 0.28 for MG, 0.57 for WS, and 0.83 for IMM. Immunohistochemical staining for H. pylori is highly specific and has a low inter observer variation. We advise that in situations where gastric histology is the main diagnostic tool, IMM should be used for the detection of H. pylori. PMID- 9354350 TI - Incidence of cytomegalovirus disease in the Aquitaine cohort of HIV-infected patients: a retrospective survey, 1987-1993. Groupe d'Epidemilogie Clinique du SIDA en Aquitaine (GECSA). AB - We estimated the incidence of the first episodes of cytomegalovirus (CMV) disease in the Aquitaine cohort of HIV-infected subjects, south-western France. Cases were retrospectively investigated using standardized definition criteria. Retinitis was confirmed by an ophthalmologist. Gastro-intestinal lesions were confirmed histologically. Encephalitis was histologically confirmed; it was considered possible if TDM or magnetic resonance imaging (MRI) and symptomatology suggested this diagnosis. Pneumopathy was probable in case of hypoxemia, interstitial X-Ray images and response to CMV treatment; it was confirmed if intranuclear inclusions were identified on biopsy or brushing specimen. In the cohort (n = 3525) followed for an average of 46 months, 158 patients had a first episode of CMV disease. The cumulative incidence was 4.5% and the incidence rate (IR) 1.2 per 100 person-years. The IR was higher for homosexuals (2.0) than for heterosexuals (1.0) and intravenous drug users (0.5). Retinitis was the most frequent site (90 cases), followed by digestive system (40), lung (three confirmed and 17 probable) and central nervous system (eight confirmed and three possible). Sixty-eight percent of the patients were at the AIDS stage when CMV disease was diagnosed, with a mean CD4 count of 42/mm3. The cumulative probability of developing CMV disease 2 years after falling below 200 CD4 lymphocytes/mm3 was 8.0%. Retinitis is by far the most common site for CMV disease. Homosexual transmission of HIV, clinical AIDS and low CD4 count are associated with the occurrence of the first episode of CMV disease. PMID- 9354351 TI - Absence of hepatitis C virus transmission but frequent transmission of HIV-1 from sexual contact with doubly-infected individuals. AB - Hepatitis C virus (HCV) is transmitted through infected blood and blood products, but evidence of other routes of transmission is less clearly understood. In a study designed to examine human immunodeficiency virus (HIV) transmission, the prevalence of HCV has also been measured. Sixty-one couples were analysed, 30 in which partners were at risk through sexual contact alone, of whom 12 (40%) became infected with HIV and none with HCV. Thirty-one partners were exposed sexually and additionally through intravenous drug use. Of these, 16 (52%) became infected with HIV and 25 (80%) contracted HCV infection. These findings support the evidence of others that HCV is only rarely transmitted by sexual intercourse in heterosexual relationships and that HIV is not a co-factor for HCV transmission. PMID- 9354353 TI - Cardiovascular disease risk from prior Chlamydia pneumoniae infection can be related to certain antigens recognized in the immunoblot profile. AB - Chlamydia pneumoniae infection has been described as a risk factor for atherosclerosis on the basis of raised seroreactivity against complete elementary bodies among cardiovascular disease (CVD) patients. In order to identify antigens of possible pathogenetic relevance, C. pneumoniae IgG and IgA immunoblot profiles were compared for CVD patients (IgG: n = 159; IgA: n = 72) and for controls (IgG: n = 158; IgA: n = 115), all with prior C. pneumoniae infection. IgG and IgA recognition patterns were very similar, and a broad range of antigens was commonly recognized. However, statistical analysis demonstrated IgG seroresponses to 40, 54, 60, 75, and 98 kDa antigens to be more frequent among patients and resulting in odds ratios between 2.3 (98 kDa) and 29.4 (40 kDa) for development of CVD. This relation remained evident after adjustment for age and sex. Cardiovascular risk from prior chlamydial infection can thus be linked to certain antigens. Thus, for the first time potential atherogenetic virulence factors of C. pneumoniae are described. Though causal relation of chlamydial and atherosclerotic disease cannot be proven yet, evidence is growing that chlamydial structures play a part in the multifactorial pathogenesis of one of the most prevalent health hazards world-wide. PMID- 9354352 TI - Strains of Alcaligenes faecalis from clinical material. AB - Six strains of Alcaligenes faecalis, unusually isolated from clinical material, are described. Alcaligenes faecalis is a Gram-negative catalase- and oxidase positive, motile rod. It is commonly found in a watery environment and is rarely isolated from humans. The clinical and laboratory characteristics of the clinical A. faecalis isolates are presented. PMID- 9354354 TI - Another cause of necrotizing fasciitis? AB - Necrotizing fasciitis is a rare condition, but has a devastating clinical course if it is not diagnosed early and treated aggressively. There are usually a number of organisms implicated in its pathogenesis, but in this case report a patient is presented in whom the only organism isolated from both tissue and blood cultures was Clostridium septicum. This organism almost always causes a myonecrosis, but in this case only the superficial and deep fascial layers were affected, sparing the underlying muscle. Clostridium septicum resulting in a true necrotizing fasciitis with no myonecrosis and being the only isolate from both blood and tissue cultures has not been previously reported in literature. PMID- 9354355 TI - Pressure ulcer accelerated healing with local injections of granulocyte macrophage-colony stimulating factor. AB - This is the first report of granulocyte macrophage-colony stimulating factor (GM CSF) inducing accelerated healing of a sacral pressure ulcer in a bedridden patient with bilateral hemiplegia. GM-CSF was diluted and injected locally around and into the ulcer bed every 2-3 days for 2 weeks, then weekly for 4 weeks until complete healing occurred. A new firm granulation tissue was noted within a few days. The ulcer showed 85% healing within 2 weeks and 100% by 2 months. Healing started from the periphery and from within the ulcer bed at sites of GM-CSF injections. It was slower at areas where there was complete necrosis and detachment of skin from underlying tissue. The ulcer remained closed until the patient's sudden death 9 months later. A biopsy of granulation tissue showed inflammatory cells and reactive fibroblasts. The potential role of GM-CSF and growth factors in pressure ulcer therapy and wound healing are discussed. PMID- 9354357 TI - Ruptured abdominal aorta secondary to psoas muscle abscess due to Klebsiella pneumoniae in an alcoholic. AB - An alcoholic patient with low back pain and Klebsiella pneumoniae septicaemia is reported. Computed tomography revealed abdominal aortic rupture associated with a psoas abscess. Aortic ligation above and below the rupture site and an axillo femoral bypass were performed, but the patient died on the first postoperative day. Alcoholism is a common underlying disease in K. pneumoniae septicaemia and its septic metastasis to the psoas muscle. The prognosis of aortic infection secondary to psoas abscess is very poor once aortic rupture occurs. Prompt abscess drainage following correct diagnosis and arterial reconstruction before aortic rupture are mandatory. PMID- 9354356 TI - Disseminated cutaneous zoster and aseptic meningitis in a previously healthy patient. AB - A previously healthy, 37-year-old immunocompetent man presented with disseminated cutaneous zoster and aseptic meningitis. Varicella zoster virus DNA was recovered from the cerebrospinal fluid (CSF) by the polymerase chain reaction. Cytological evaluation of the CSF revealed 'reactive, highly atypical lymphocytosis'. The patient fully recovered after treatment with aciclovir. PMID- 9354358 TI - Systemic and local immune responses of four cases with lower respiratory tract illness due to reinfection with respiratory syncytial virus. AB - Four infants and children who had tracheobronchitis or bronchiolitis by respiratory syncytial virus (RSV) reinfection were presented. Their clinical features, systemic and local immune reactions to RSV were compared with those of eleven age-matched patients who had lower respiratory tract illness with primary RSV infection. Moderate antibody activities in their nasopharyngeal secretions and sera which were comparable to those of convalescent phase of primary infection were observed within several days after onset of illness; however, their clinical symptoms, such as fever or wheezing, lasted almost as long as in primary RSV infection. These observations suggest that the higher serum and secretory antibody activity to RSV during acute phase does not always bring about clinical amelioration in RSV reinfection. The contribution of immunopathological reaction between RSV and RSV-specific antibodies might also be considered as a cause of inflammation during the acute phase of RSV reinfection. PMID- 9354359 TI - Brucellar osteomyelitis involving prosthetic extra-articular hardware. AB - We report the case of a patient with Brucella melitensis osteomyelitis involving non-joint prosthetic implant of the femur. This is the first case published of osteomyelitis by Brucella sp. in a patient with prosthetic implant in bone and the second one with both intra- or extra-articular prosthetic bone implant. Brucella melitensis is a rare organism which causes osteomyelitis in patients with prosthetic hardware, and should be added to the list of suspected organisms responsible for this disease, especially in endemic areas of brucellosis. PMID- 9354360 TI - Sporadic case of listeriosis associated with the consumption of a Listeria monocytogenes-contaminated 'Camembert' cheese. AB - Listeria monocytogenes is an intracellular gram-positive organism responsible for severe infections in both humans and animals. Whereas the food-borne transmission of listeriosis was demonstrated in several outbreaks, most cases of listeriosis occur sporadically and are rarely linked with consumption of contaminated foods. In this paper a case of septicaemia with L. monocytogenes in a 73-year-old immunocompromised man is described. Evidence for the association of this case of listeriosis with the consumption of a contaminated 'Camembert' cheese is provided by serotyping, esterase typing, DNA macrorestriction patterns analysis and level of virulence of the isolated strains for mice. PMID- 9354361 TI - Penicillin-resistant Streptococcus pneumoniae isolated in Barbados. AB - An imported case of pneumonia caused by penicillin-resistant Streptococcus pneumoniae occurred in a tourist, shortly after arriving in Barbados. The isolate was of serogroup 6 and exhibited intermediate resistance to penicillin. This was the first isolation of penicillin-resistant S. pneumoniae in Barbados. PMID- 9354362 TI - Evidence of Chlamydia pneumoniae infection obtained by the polymerase chain reaction (PCR) in patients with acute myocardial infarction and coronary heart disease. PMID- 9354363 TI - Comparative evaluation of naturally occurring Pneumocystis carinii pneumonia (PCP) and PCP despite primary chemoprophylaxis in patients with AIDS. PMID- 9354364 TI - Corynebacterium afermentans spp. afermentans sepsis in a neurosurgical patient. PMID- 9354365 TI - Co-infection with several HCV genotypes enhances liver damage in patients with chronic hepatitis C. The Hepatitis/HIV Spanish Study Group. PMID- 9354366 TI - Polymicrobial septicaemia with Pseudomonas aeruginosa and Streptococcus pyogenes following traditional tattooing. PMID- 9354367 TI - Specific interaction between erythrocytes and a glucose-carrying polymer mediated by the type-1 glucose transporter (GLUT-1) on the cell membrane. AB - A reducing glucose-carrying polymer, called poly [3-O-(4'-vinylbenzyl)-D-glucose] (PVG), interacted with erythrocytes carrying the type-1 glucose transporter (GLUT 1) on the cell membrane. The cooperative interaction between a number of GLUT-1s and a number of reducing 3-O-methyl-D-glucose moieties on a PVG polymer chain is responsible for the increase in the interaction with erythrocytes. In contrast to the PVG homopolymer, other sugar-carrying polymers showed lower interaction with erythrocytes. The affinity of erythrocytes and PVG was studied using FITC-labeled glycopolymers. The fluorescence intensity significantly changed, whereas a small change in fluorescence intensity was observed for other homopolymers. The specific interaction between GLUT-1 on erythrocytes and the PVG polymer carrying reducing glucose was suppressed by the inhibitors, phloretin, phloridzin, and cytochalasin B, and a monoclonal antibody to GLUT-1. Direct observation by confocal laser microscopy with the use of FITC-labeled PVG demonstrated that erythrocytes interacted with the soluble form of the PVG polymer via GLUT-1, while fluorescence labeling of the cell surface was prevented on pretreatment with the monoclonal antibody to GLUT-1. PMID- 9354368 TI - The two lectin domains of the tandem-repeat 32-kDa galectin of the nematode Caenorhabditis elegans have different binding properties. Studies with recombinant protein. AB - Some properties of recombinant proteins derived from the 32-kDa galectin isolated from the nematode Caenorhabditis elegans, which lectin is composed of two tandemly repeated homologous domains [Hirabayashi et al. (1992) J. Biol. Chem. 267, 15485], were studied in order to elucidate the function of this unique polypeptide architecture. We expressed the whole molecule (N32), the N-terminal lectin domain (Nh), and the C-terminal lectin domain (Ch) in Escherichia coli using the expression vector pET21a. All of the recombinant proteins were bound by asialofetuin-Sepharose. CD spectra of the recombinant proteins indicated all of them to be rich in beta-structure and properly refolded. Gel filtration on an HPLC column suggested that all of them existed as monomers. Neither Nh nor Ch seemed to form dimers, in contrast to vertebrate proto-type galectins. Only N32 showed hemagglutination activity towards trypsinized rabbit erythrocytes. Comparison of the affinity of N32, Nh, and Ch for asialofetuin-Sepharose by frontal affinity chromatography [Kasai et al. (1986) J. Chromatogr. 376, 33] showed that Ch has 7-fold weaker affinity than N32, and Nh proved to have still weaker affinity. Since the Asn residue in the CRD (carbohydrate recognition domain), which is conserved in all other galectins, is substituted by Ser in the case of Nh, these data suggest that the two CRDs in this tandem-repeat galectin have different sugar binding properties and that the 32-kDa galectin may serve as a heterobifunctional crosslinker. PMID- 9354369 TI - Pepsinogens and pepsins from house musk shrew, Suncus murinus: purification, characterization, determination of the amino-acid sequences of the activation segments, and analysis of proteolytic specificities. AB - Three pepsinogens, namely, pepsinogens A, C-1, and C-2, were purified from gastric mucosa of adult house musk shrew (Suncus murinus) by conventional chromatographic and gel filtration procedures. The molecular masses were 40, 39, and 41 kDa for pepsinogens A, C-1, and C-2, respectively. Pepsinogen C-2 contains an Asn-linked carbohydrate chain(s) of about 2 kDa. Each pepsinogen was converted to pepsin through an intermediate form under acidic conditions. By NH2-terminal sequence analysis of these protein species, the amino acid sequences of activation segments (proparts) of pepsinogens A and C-1 were determined to be LYKVPLVKKKSLRQNLIENGLLKDFLAKHNVNPASKYFPTE and KVTKVTLKKFKSIRENLREQGLLEDFLKTNHYDPAQKYHFGDF, respectively. The similarity of these two sequences is nearly 50%. Each pepsin cleaved preferentially peptide bonds between hydrophobic and aromatic amino acids, or bonds on either side of these amino acids. Although each activation segment had several sites susceptible to pepsin action, activation proceeded by limited cleavages of the segment, presumably due to the steric inflexibility of the segment in native pepsinogen. The activity of pepsin A was inhibited completely in the presence of a more than equimolar amount of pepstatin, while a hundred-molar excess amount of pepstatin was needed for the complete inhibition of the activity of pepsins C-1 and C-2. PMID- 9354370 TI - Role of Ca2+-independent phospholipase A2 in exocytosis of amylase from parotid acinar cells. AB - We evaluated the role of cytosolic phospholipase A2 (PLA2) in the exocytosis of amylase from parotid acinar cells. The exocytosis stimulated by isoproterenol was dose-dependently inhibited by bromoenol lactone (BEL), a potent suicide inhibitor of Ca2+-independent cytosolic PLA2. The IC50 value of BEL was approximately 7 microM. AACOCF3, a selective inhibitor of Ca2+-dependent cytosolic PLA2, did not inhibit the exocytosis at least up to 30 microM. BEL also inhibited amylase release evoked by forskolin and membrane-permeable cAMP, but it did not inhibit cAMP-dependent protein kinase activity. PLA2 activity in parotid acinar cells was found to be predominantly Ca2+-independent, and was strongly inhibited by BEL, whose IC50 value was approximately 2 microM when it was applied to intact acini. Although isoproterenol scarcely enhanced [3H]arachidonic acid release from intact acinar cells, BEL dose-dependently decreased the basal arachidonic acid release to approximately one half of the control value. These results suggest that the cytosolic Ca2+-independent PLA2 activity plays a role in the membrane fusion process of exocytosis in parotid acinar cells. PMID- 9354371 TI - Nucleotide sequence of gene PBI encoding a protein homologous to salivary proline rich protein P-B. AB - The nucleotide sequence of gene PBI, the putative translational product of which is homologous to salivary proline-rich protein P-B, has been determined. PBI is 6.4 kb long and contains 3 exons. PBI appears to code for the precursor of a proline-rich protein which we have designated P-B1. The P-B1 precursor is composed of 134 amino acid residues including 22 residues of signal sequence, 23 residues of N-terminal sequence, five repeating units of 13 to 14 residues and 22 residues of C-terminal sequence. The signal sequence of this precursor is identical with that of the P-B precursor. The N-terminal 61 residue sequence of P B1 has homology of 75% with the whole sequence of P-B (57 residues), when deletion of 4 residues is taken into account. P-B1 is 55 residues longer than P B. PMID- 9354372 TI - Screening of a mutant plasmid with high expression efficiency of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus, in Escherichia coli. AB - A mutant plasmid with elevated expression efficiency of GC-rich Thermus thermophilus leuB gene was screened in Escherichia coli. A wild-type plasmid pHB2 carrying T. thermophilus leuB gene was introduced into leuB-deficient E. coli C600 cells. During successive cultures of the transformant in leucine-free medium, the original plasmid was spontaneously replaced by a mutant plasmid. The expression efficiency of the leuB gene on the mutant plasmid was 4.8-fold higher than that of the wild-type plasmid. Sequencing of the mutant plasmid revealed that the open reading frame (ORF1) in front of the leuB gene was shortened from 822 to 306 bp. Several expression vectors were constructed to investigate the effect of the length of ORF1, and the optimal length for the expression of the following leuB gene was determined. It was also shown that the stop codon of ORF1 should be overlapped with the initiation codon of leuB gene for the highest efficiency. PMID- 9354373 TI - Inverted repeats in the TATA-less promoter of the rat catalase gene. AB - The rat catalase gene promoter lacks a TATA box, and has eight initiation sites of transcription as well as several GT and CCAAT boxes. To elucidate the mechanism of transcription in this TATA-less promoter, we analyzed nuclear factors binding to this regulatory region of the catalase gene. Functional analysis of the promoter region revealed that a pair of inverted repeat motifs located on both sides of the transcription initiation sites played an important role in the gene expression. The core sequence of this element is GYCMGGCCCKCTCYKG (M=A/C, K=G/T, Y=T/C), and four species of complex were observed to bind to this sequence. Furthermore, this element in the chimeric promoter negatively interacted with the initiator element of the terminal deoxynucleotidyl transferase gene. These results suggested that the rat catalase gene is regulated by a novel mechanism involving the inverted repeated structure of the promoter and characteristic binding factors. PMID- 9354374 TI - Nitric oxide-induced modification of glyceraldehyde-3-phosphate dehydrogenase with NAD+ is not ADP-ribosylation. AB - One biological effect of nitric oxide (NO) has been believed to be exerted through induction of the ADP-ribosyltransferase activity of glyceraldehyde-3 phosphate dehydrogenase (GAPDH). Though this notion is based on the finding that NO increases the auto-ADP-ribosylation of GAPDH, controversial data have also been reported. To determine whether or not NO really activates ADP-ribosylation, we re-examined the NO-induced modification of GAPDH with NAD+. GAPDH was modified equally with [adenosine-14C]NAD+ and [carbonyl-14C]NAD+, indicating that the glycoside bond of NAD+ between ADP-ribose and nicotinamide is intact. The release of nicotinamide from NAD+ was not evident during incubation of GAPDH with [carbonyl-14C]NAD+. Thus, the modification of GAPDH is apparently not ADP ribosylation. In addition, we found that basal and glyceraldehyde-3-phosphate induced modifications of GAPDH, both of which have also been explained as ADP ribosylation, were not ADP-ribosylation, and that the modification of GAPDH in the absence and presence of NO or GA3P was distinct in the dithiothreitol effect or resistance to HgCl2. PMID- 9354375 TI - Characterization of tyrosine-phosphorylated delta isoform of protein kinase C isolated from Chinese hamster ovary cells. AB - Phorbol ester treatment of Chinese hamster ovary cells stably overexpressing the delta isoform of protein kinase C induced the association of the isoform with the particulate fraction and the tyrosine phosphorylation of a small portion of the delta isoform. The delta isoform without tyrosine phosphorylation was recovered as an enzyme dependent on phospholipid and diacylglycerol, whereas the tyrosine phosphorylated delta isoform was recovered in two fractions, one dependent on, and the other independent of, phospholipid and diacylglycerol. The tyrosine phosphorylated delta isoform independent of lipid activators might be associated with phorbol ester and phospholipids. Immunoblot analysis revealed that the delta isoform is a doublet protein of 76 and 78 kDa, and that the delta isoform fraction without tyrosine phosphorylation contained 76- and 78-kDa proteins, whereas the tyrosine-phosphorylated delta isoform contained the 78-kDa protein but not the 76-kDa protein. In vitro analysis showed that the 78-kDa protein of the delta isoform without tyrosine phosphorylation is an efficient substrate of tyrosine kinase only when phosphatidylserine and either diacylglycerol or phorbol ester are present; however, the 76-kDa protein can not be tyrosine-phosphorylated even in the presence of these lipid activators. The phospholipid and diacylglycerol-dependent form of the tyrosine-phosphorylated enzyme isolated from the cell line required lower concentrations of phosphatidylserine and phorbol ester for its activity in vitro as compared with the enzyme without tyrosine phosphorylation. These results suggest that the tyrosine-phosphorylated enzyme generated upon stimulation of the cells may associate with membranes and exert its full activity even with the lower concentrations of the lipid activators. PMID- 9354376 TI - Effect of unsaturated fatty acids and alpha-tocopherol on immunoglobulin levels in culture medium of rat mesenteric lymph node and spleen lymphocytes. AB - Mesenteric lymph node (MLN) and spleen lymphocytes of Sprague-Dawley rats were cultured with 1 mM unsaturated fatty acids (UFAs) with or without 100 microM alpha-tocopherol (Toc), and the immunoglobulin content and thiobarbituric acid (TBA) value of the culture media were measured to clarify the relationship between lipid peroxidation and the IgE level in the culture medium. The increase in the IgE content and TBA value induced by UFAs was alleviated in the presence of Toc in both lymphocytes, and was correlated well with their oxidation rates in most cases. Gamma-linolenic acid enhanced the IgE level much more than would be expected from its oxidation rate in both lymphocytes, and linoleic acid showed similarly high activity only in splenocytes. These results suggest that lipid peroxidation is partly responsible for the enhancement of IgE level induced by UFAs. PMID- 9354377 TI - Formation of recombinant human procollagen I heterotrimers in a baculovirus expression system. AB - The present study describes the production of human procollagen I in a baculovirus expression system. Recombinant baculovirus carrying pro alpha1(I) or pro alpha2(I) cDNA was constructed and infected to Sf9 cells. Full-length pro alpha1(I) or pro alpha2(I) chains were synthesized by the cells infected with either of the recombinant viruses. The pro alpha1(I) chains formed pepsin resistant homotrimers stabilized by interchain disulfide bonds, a small proportion of which was secreted into the culture medium. The pro alpha2(I) chains were not linked into trimers by disulfide bonds and failed to form stable triple helices, although some chains were suggested to exist as dimers or unstable trimers in which only two chains were linked by disulfide bonds. In spite of their non-helicity, the pro alpha2(I) chains were secreted at a higher rate than the pro alpha1(I) chains. Sf9 cells simultaneously synthesized both pro alpha1(I) and pro alpha2(I) chains when the cells were co-infected with the two recombinant viruses. Pepsin-treatment of the product clearly demonstrated the production of procollagen I heterotrimers composed of two pro alpha1(I) chains and one pro alpha2(I) chain, homotrimers of the pro alpha1(I) chains being negligible. This expression system appears to offer a unique means of studying the mechanism of chain association and secretion during procollagen biosynthesis. PMID- 9354378 TI - Dissociation of non-complementary second DNA from RecA filament without ATP hydrolysis: mechanism of search for homologous DNA. AB - RecA protein catalyzes the DNA annealing and mimics the DNA strand exchange reaction in vitro in the presence of ATP or its non-hydrolyzable analog, adenosine 5'-O-3-thiotriphosphate (ATPgammaS). For these activities RecA coordinates two DNA molecules [Takahashi, M. and Norden, B. (1994) Adv. Biophys. 30, 1-35]. In order to get a better understanding of how RecA performs the search for sequence complementarity or homology between two DNA molecules, the association and dissociation kinetics of a second DNA molecule to and from RecA in the presence of ATPgammaS have been investigated. The kinetics were monitored by fluorescence measurements of partly etheno-modified poly(dA) assisted by linear dichroism measurements of the flow-oriented complex. The association of the second DNA is fast, regardless of whether the sequence is complementary or not. By contrast, the dissociation kinetics is strongly dependent on sequence complementarity. If the second DNA is complementary to the first, dissociation is extremely slow, whilst that of non-complementary second DNA is fast. In no case does the first DNA leave the RecA fiber. Our findings indicate that the dissociation step is important in the search for homology by RecA. PMID- 9354379 TI - Analysis of the stabilization of hen lysozyme by helix macrodipole and charged side chain interaction. AB - In the N-terminal region of the alpha-helix of the c-type lysozymes, two Asx residues exist at the 18th and 27th positions. Hen lysozyme has Asp18/Asn27 (18D/27N), and we prepared three mutant lysozymes, Asn18/Asn27 (18N/27N), Asn18/Asp27 (18N/27D), and Asp18/Asp27 (18D/27D). The stability of the wild-type (18D/27N) lysozyme supported the existence of a hydrogen bond between the side chain of Asp18 and the amide group at the N1 position in the alpha-helix, while the stability of the 18N/27D lysozyme supported the presence of the capping box between the Ser24 (N-cap) and Asp27 residues. Although electrostatic repulsion was observed between Asp18 and Asp27 residues in 18D/27D lysozyme, the dissociation of each residue contributed to stabilizing the B-helix in 18D/27D lysozyme through hydrogen bonding and charge-helix macrodipole interaction. This is the first evidence that two neighboring negative charges at the N-terminus of the helix both increased the stability of the protein. PMID- 9354380 TI - Characterization and regulation of taurine transport in Caco-2, human intestinal cells. AB - We characterized the taurine transport system in human intestinal Caco-2 cells and showed that it is subject to adaptive regulation. The activity of taurine transport in Caco-2 cells was evaluated by means of an Na+- and Cl(-)-dependent high-affinity transport system, the characteristics of which were similar to those of the beta-amino acid-specific taurine transport system previously described for various tissues. The activity of taurine transport was down regulated on culturing in taurine-containing medium. This taurine-induced down regulation was dependent on both the incubation time with taurine and the concentration of taurine. Hypotaurine and beta-alanine were also capable of inducing this adaptive regulation, whereas alpha-amino acids and gamma aminoisobutyric acid were not. Kinetic analysis of control and taurine-treated cells suggested that the down-regulation was associated with a decrease in the maximal velocity of taurine transport and also with a decrease in the affinity of the taurine transporter. Cycloheximide treatment weakened the taurine-induced down-regulation. The mRNA level of the taurine transporter (HTAU type) in taurine treated cells was markedly decreased compared with in control cells. These results indicate that a complex regulatory mechanism is involved in this down regulation. PMID- 9354381 TI - Identification of reactive site of a proteinaceous metalloproteinase inhibitor from Streptomyces nigrescens TK-23. AB - Streptomyces metalloproteinase inhibitor (SMPI), isolated from Streptomyces nigrescens TK-23, is a small proteinaceous metalloproteinase inhibitor consisting of 102 amino acid residues and two disulfide bridges. SMPI specifically inhibits metalloproteinases such as thermolysin. After prolonged incubation with a catalytic amount of thermolysin, it is cleaved at Cys64-Val65 [Murai, H., Hara, S., Ikenaka, T., Oda, K., and Murao, S. (1985) J. Biochem. 97, 173-180]. Hence, for identification of the reactive site, mutants were constructed by substituting Val65 with various amino acid residues (Leu, Ile, Phe, Tyr, Gly, Ser, Lys, and Glu). The mutants were analyzed for inhibitory activity. Among them, V65I, V65L, V65F, and V65Y retained strong inhibitory activity, whereas V65S, V65G, V65K, and V65E showed very weak inhibitory activity against thermolysin. The Ki values were found to be of the order of 10(10) M by using a fluorogenic substrate, MOCAc-Pro Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2. In addition, susceptibility to enzyme degradation was analyzed by means of limited proteolysis with thermolysin. Mutants which retained strong inhibitory activity were cleaved by thermolysin only at the reactive site, in the same way as native SMPI. The mutants which showed weak inhibitory activity underwent rapid degradation. These results were consistent with the substrate specificity of thermolysin. Based on these results, the reactive site of SMPI was identified as Cys64-Val65. PMID- 9354382 TI - A simple and rapid purification procedure minimizes spontaneous oxidative modifications of low density lipoprotein and lipoprotein (a). AB - Usual purification procedures of LDL and Lp(a) require numerous, extensive and prolonged sample handlings: this greatly increases the possibility of spontaneous oxidation. We have developed a method which, making use of two short-run ultracentrifugations in vertical rotors alternated by two rapid column chromatography steps (SRUC), significantly shortens the preparation time to 3.5 h (LDL) and does not demand additional instrumentation or particular accuracy. Purification of Lp(a) requires a further wheat germ agglutinin chromatographic step, which can be accomplished within 30 min. More importantly, the method significantly reduces spontaneous oxidation as compared with classical isolation procedures. LDL isolated by the standard sequential method exhibits more extensive apolipoprotein B100 degradation, lipid peroxidation, and endogenous antioxidant (vitamin E) loss than the same lipoproteins obtained by means of the SRUC. This procedure may have be particularly valuable in experiments evaluating the effects of oxygen radical-induced modifications, especially in vitro. PMID- 9354383 TI - Purification and characteristics of recombinant mouse metallothionein-I from Escherichia coli. AB - The mouse metallothionein-I (mMT-I) cDNA was amplified by polymerase chain reaction (PCR), inserted into vector pGEX-4T-1, and expressed in Escherichia coli as a carboxyl terminal extension of the 26-kDa glutathione-S-transferase (GST). Analyzed by SDS-PAGE, the amount of the expressed fusion protein GST-MT was over 50% of total cellular proteins. After the fusion protein had been digested with thrombin on a Glutathione-Sepharose 4B affinity chromatography column, recombinant mMT-I was purified by gel filtration on Sephadex G50. The results of molecular mass, amino acid composition and sequence of 10 amino acids at the N terminus of the recombinant mMT-I demonstrate that the purified protein is the one we desired. The ratios of metal:protein and thiol:protein are the same as those of wild-type MT. The half-dissociation pHs of Cd, Cu, and Zn from recombinant mMT-I were 3.57, 1.40, and 5.20, respectively, which are in agreement with those from native rabbit MT-I. The ultraviolet absorbance and circular dichroism (CD) spectra at pH 8.0 and pH 2.0 were all similar to those of native MT, indicating that they have the same metal-thiolate structure even though six amino acid residues have been added at the N-terminus of the recombinant protein. PMID- 9354384 TI - Purification and characterization of diacylglycerol acyltransferase from the lipid body fraction of an oleaginous fungus. AB - Diacylglycerol acyltransferase (DGAT) [EC 2.3.1.20] was purified to apparent homogeneity from the lipid body fraction of an oleaginous fungus, Mortierella ramanniana var. angulispora. The enzyme was solubilized from the lipid body fraction with 0.1% Triton X-100, and purified by subsequent column chromatography on Yellow 86 agarose, Superdex-200, Heparin-Sepharose, second Superdex-200, and second Yellow 86 agarose. The enzyme activity was finally enriched 4,802-fold over that of the starting 1,500X g supernatant. The apparent molecular mass of the enzyme was 53 kDa on SDS polyacrylamide gel electrophoresis. The purified enzyme did not exhibit glycerol-3-phosphate acyltransferase, lysophosphatidic acid acyltransferase, lipase, transacylase, or acyl-CoA hydrolase activities, although 2-monoolein was acylated with about a half of the enzyme activity toward 1,2-diolein. The purified DGAT depended on exogenous sn-1,2-diolein and oleoyl CoA, with the highest activity at about 200 and 20 microM, respectively. Purified DGAT utilized a broad range of molecular species of both diacylglycerol and acyl CoA as substrates. The highest activity was observed with sn-1,2-diolein and lauroyl-CoA. Anionic phospholipids such as phosphatidic acid (PA) activated the purified enzyme, as found for the Triton X-100 extract. Sphingosine dose dependently inhibited DGAT activity activated by PA and basal activity without PA. These results provide a basis for further studies on the molecular mechanism of triacylglycerol biosynthesis and lipid body formation, in which DGAT plays an important role. PMID- 9354385 TI - Assignment of imino proton signals of G-C base pairs and magnesium ion binding: an NMR study of bovine mitochondrial tRNA(SerGCU) lacking the entire D arm. AB - The mammalian mitochondrial tRNA(SerGCU) (mt tRNA[SerGCU]) has a unique structure in that it lacks the whole D arm. To elucidate its higher-order structure, we synthesized unmodified bovine mt tRNA(SerGCU) using T7 RNA polymerase and measured its 1H-NMR spectrum in the imino proton region. Although the imino proton signals heavily overlapped, we succeeded in assigning all the seven imino proton signals of the G-C base pairs by a combination of base replacement and 15N labeling of the G residues of a whole tRNA molecule or of the 3'-half fragment. The results indicate that the tRNA possesses the secondary structure that has been supposed on the basis of biochemical studies. Analysis of the effect of the magnesium concentration on the G-C pairs suggests that the acceptor and T stems do not form a co-axial helix, and that the core region of the tRNA does not interact with magnesium ions. These features are significantly different from those of canonical tRNAs. Despite this, it is very likely that the tRNA as a whole takes a nearly L-shape tertiary structure. PMID- 9354386 TI - Properties of hepatitis delta virus ribozyme, which consists of three RNA oligomer strands. AB - Properties of a hepatitis delta virus (HDV) RNA ribozyme system, which consists of three RNA oligomer strands (substrate 8-mer; enzyme 16-mer plus 35-mer) and contains a hybrid sequence of genomic and antigenomic RNA cores, are reported. Effects of Mg2+ concentration, divalent metal ion species, pH, and temperature on the cleavage activity were examined. The substrate cleavage activity increased with increasing Mg2+ concentration (0-100 mM). Ca2+ and Mn2+ ions were the most effective divalent cations and Mg2+ was less effective. The cleavage activity increased with increasing pH (5-7.5). The optimum temperature for the cleavage activity was 25-40 degrees C. The Mg2+ concentration, pH and temperature dependencies are different from those reported for the single-strand ribozymes (about 90-mer) although the divalent metal ion preference is very similar. Conformational change induced by Mg2+ ion titration was monitored by CD. The CD data and the activity-Mg2+ concentration data were analyzed by curve-fitting analysis using equations derived for multiple metal ion binding mechanisms. The data can be explained by a model in which three Mg2+ ions bind to one ribozyme unit. PMID- 9354387 TI - Chemical structure of lipid A from Helicobacter pylori strain 206-1 lipopolysaccharide. AB - The chemical structure of a novel lipid A, which was obtained as a major component from lipopolysaccharide of Helicobacter pylori strain 206-1, was determined to be a glucosamine beta(1-6) disaccharide 1-(2-aminoethyl)phosphate acylated by (R)-3-hydroxyoctadecanoic acid and (R)-3 (octadecanoyloxy)octadecanoic acid at the 2- and 2'-position, respectively. The absence of a phosphoryl group at the 4'-position and fatty acyl groups at the 3- and 3'-position, and the stoichiometric presence of 2-aminoethyl phosphate at the 1-position are unique features, distinguishing it from the lipid A of enterobacteria. PMID- 9354388 TI - Preparation of antibodies highly specific to N1,N8-diacetylspermidine, and development of an enzyme-linked immunosorbent assay (ELISA) system for its sensitive and specific detection. AB - N8-Acetylspermidine was coupled to mercaptosuccinylated BSA using a bifunctional cross-linker, N-(4-maleimidobutyryloxy)succinimide, and the resulting conjugate was used to raise N1,N8-diacetylspermidine (DiAcSpd)-specific antibodies in rabbits. DiAcSpd-specific antibodies were enriched from crude sera through a series of affinity-based fractionations using ligands with structures mimicking those of DiAcSpd and monoacetylspermidines. With the N8-acetylspermidine-BSA conjugate as a solid phase antigen in a competitive ELISA system, the selectivity for DiAcSpd over other polyamine species was high, but competition by DiAcSpd added to the fluid phase was too weak for the system to be applicable to measurement of the concentration of DiAcSpd in human urine. In contrast, with the N1-acetylspermidine-BSA conjugate adsorbed on the ELISA plate, DiAcSpd efficiently competed for the same antibody, thus yielding a sensitive competitive ELISA system for measuring DiAcSpd. The Ki value for DiAcSpd with the latter competitive ELISA system was 54 nM, and the cross-reactivity with DiAcSpd, N1,N12 diacetylspermine, N8-acetylspermidine, N1-acetylspermidine, and acetylputrescine was 100, 1.2, 0.74, 0.12, and 0.08%, respectively. The DiAcSpd-specific antibodies and the competitive ELISA system developed in this study will prove to be useful for analyzing the urinary level of DiAcSpd, that was recently shown to be a promising diagnostic and prognostic indicator of malignant disorders. PMID- 9354389 TI - Spectroscopic studies of rat liver acyl-CoA oxidase with reference to recognition and activation of substrate. AB - Two forms of rat peroxisomal acyl-CoA oxidase (ACO-I and -II) interact with the substrate analogs, 3-ketoacyl-CoAs, forming a complex characterized by the so called charge-transfer (CT) band around 575 nm in the absorption spectra. The CT band of ACO-I exhibited a broad dependency on the acyl chain-length from C4 to C16, whereas that of ACO-II showed increased intensity with a longer acyl chain to reach a maximum with a chain-length of C12. These chain-length dependencies of the CT bands were compared with those of the enzymatic activities reported previously [Setoyama et al. (1995) Biochem. Biophys. Res. Commun. 217, 482-487]. The differences in spectroscopic and enzymatic properties between ACO-I and -II suggest that the amino acid stretch corresponding to the third exon in the ACO sequence affects the binding of the ligand and substrate, since the difference in the primary structure between ACO-I and -II lies in the short amino acid stretch corresponding to the third of the total of 14 exons. On the other hand, resonance Raman spectra of the complexes of ACO-I and -II with 3-ketoacyl-CoAs excited in the CT band showed similar features. The two prominent FAD bands II and III, associated with the C(4a)=N(5) moiety of FAD, were observed at 1,577 and 1,545 cm(-1), respectively. In contrast, the bands at 1,615 and 1,493 cm(-1) in the ACO I x 3-keto-C8-CoA complex were assigned to the stretching modes of C=O at positions 3 and 1 of the ligand, respectively, by using the isotopically labeled ligands. Both C=O stretching bands were shifted to lower wave numbers upon complex formation with ACO-I, implying that the C=O bond involves the single bond (C-O-) character in the active site cavity. The downshift of the C(1)=O stretching band was larger than that of the C(3)=O stretching band. Therefore, the ligand lies in the active site as the anionic form with a major contribution from C(1)-O-. These observations demonstrate that the CT band around 575 nm arises from the charge-transfer interaction between the oxidized FAD and the enolate transformed after the elimination of the a-proton. The band II of FAD in the complexes reveals a significant decrease in the frequency in comparison with the complexes of medium-chain acyl-CoA dehydrogenase (MCAD) with 3-ketoacyl-CoA. This observation suggests a difference between ACO and MCAD in the hydrogen bonding network associated with enzyme-bound FAD. PMID- 9354390 TI - A Raman study on the C(4)=O stretching mode of flavins in flavoenzymes: hydrogen bonding at the C(4)=O moiety. AB - Raman spectroscopy was used to investigate the hydrogen bonding at the C(4)=O moiety of the isoalloxazine nucleus in a series of flavins and flavoproteins. Isotope effects of Raman bands confirmed that the band observed around 1,710 cm( 1) is mainly derived from C(4)=O stretching vibrational mode. A linear correlation was observed between the frequency of C(4)=O stretching and the chemical shift of 13C(4), suggesting that the data from both Raman and NMR spectroscopies reflect a common perturbation, i.e., hydrogen bonding. The maximum difference of C(4)=O frequency among flavins and flavoproteins examined is 36 cm( 1) [1,723 cm(-1) for riboflavin-binding protein (Kim, M. and Carey, P.C. (1993) J. Am. Chem. Soc. 115, 7015-7016) and 1,687 cm(-1) for the complex of medium chain acyl-CoA dehydrogenase with acetoacetyl-CoA]; the maximum difference of 40 70 kJ/mol in the hydrogen bonding strength at the C(4)=O exists among flavoproteins. By use of an empirical linear correlation between the frequency of C=O stretching and the bond length of the C=O, it is estimated that the maximum difference in the bond length among flavoproteins treated here is ca. 0.017 A. The hydrogen bonding at the C(4)=O in medium-chain and short-chain acyl-CoA dehydrogenases becomes stronger upon complexation with substrate analogs. Since the hydrogen bonding at the C(4)=O is expected to enhance the electron-accepting capacity of the N(5) position, substrate-binding itself probably raises the reactivity of flavin, through enhancing the hydrogen bonding. PMID- 9354391 TI - Cloning and expression of the glutamate racemase gene of Bacillus pumilus. AB - A glutamate racemase gene (murI) was found in Bacillus pumilus cells and cloned into Escherichia coli WM335, a D-glutamate auxotroph, by means of a genetic complement method. MurI of B. pumilus encodes a 272-amino acid protein with an unusual initiation codon, TTG. The deduced amino acid sequence shows significant similarity with those of glutamate racemases from E. coli (ratio of identical residues, 28%), Pediococcus pentosaceus (44%), and Staphylococcus haemolyticus (49%). B. pumilus MurI was expressed as a fusion protein connected to the N terminal 12 residues of beta-galactosidase; the fusion protein showed glutamate racemase activity, and resembled the enzyme of P. pentosaceus in physicochemical and enzymological properties. PMID- 9354392 TI - Possible implications of the induction of human heme oxygenase-1 by nitric oxide donors. AB - To explore the involvement of nitric oxide (NO) in the induction of heme oxygenase-1, an essential enzyme in heme catabolism, we studied the effects of NO donors on the expression of heme oxygenase-1 mRNA in HeLa human cervical cancer cells. Treatment with each of three NO donors, sodium nitroprusside, 3 morpholinosydnonimine, and S-nitroso-L-glutathione, caused noticeable increases in the expression levels of heme oxygenase- mRNA, but not heme oxygenase-2 mRNA. On the other hand, nitrite or 8-bromo cGMP exerted no noticeable effect on the levels of heme oxygenase-1 mRNA. We showed that sodium nitroprusside also increased the levels of heme oxygenase-1 protein. The sodium nitroprusside mediated increase in heme oxygenase-1 mRNA levels was abolished by treatment with actinomycin D. The expression levels of heme oxygenase-1 mRNA were also increased by NO donors in human melanoma and neuroblastoma cell lines. Thus, the observed induction of heme oxygenase-1 may represent an important response to NO or NO related oxidative stress. The half lives of heme oxygenase-1 and heme oxygenase-2 mRNAs were estimated to be about 3.2 h and more than 5 h, respectively. PMID- 9354393 TI - Caenorhabditis elegans cDNA for a Menkes/Wilson disease gene homologue and its function in a yeast CCC2 gene deletion mutant. AB - The full-length cDNA coding for a putative copper transporting P-type ATPase (Cu2+-ATPase) was cloned from Caenorhabditis elegans. The putative Cu2+-ATPase is a 1,238-amino acid protein, and highly homologous to the Menkes and Wilson disease gene products mutations of which are responsible for human defects of copper metabolism. The Saccharomyces cerevisiae mutant with a disrupted CCC2 gene (yeast Menkes/Wilson disease gene homologue) was rescued by the cDNA for the C. elegans Cu2+-ATPase but not by the cDNA with an Asp-786 (an invariant phosphorylation site) to Asn mutation, suggesting that the C. elegans Cu2+-ATPase functions as a copper transporter in yeast. The expressed C. elegans protein was detected in yeast vacuolar membranes by immunofluorescence microscopy. The yeast expression system may facilitate further studies on copper transporting P-type ATPases. PMID- 9354394 TI - Nuclear localization and involvement in DNA synthesis of Sarcophaga prolyl endopeptidase. AB - A specific prolyl endopeptidase (PEP) inhibitor, ZTTA, selectively inhibited DNA synthesis by imaginal discs and cultured embryonic cells of Sarcophaga peregrina (flesh fly). PEP was found to be localized in restricted nuclear regions. Unfertilized eggs were shown to contain a maternal message for PEP and analysis of Sarcophaga embryos at blastdermal stage revealed PEP was localized exclusively in the nuclei. These results suggest that PEP participates in DNA synthesis by, and therefore cell proliferation, of insect cells. This is the first demonstration of a biological function of PEP. PMID- 9354395 TI - The transcriptional activators of the PHO regulon, Pho4p and Pho2p, interact directly with each other and with components of the basal transcription machinery in Saccharomyces cerevisiae. AB - The transcriptional regulators Pho4p and Pho2p are involved in transcription of several genes in the PHO regulon of Saccharomyces cerevisiae. Genetic evidence with temperature-sensitive pho4 and pho2 mutants suggested that Pho4p and Pho2p interact with each other. Immunoprecipitation experiments showed that Pho4p and Pho2p form a complex on a 36-bp sequence bearing an upstream activation site (UAS) and protein binding assays indicated that these proteins interact directly. DNA-binding experiments with crude extracts prepared from yeast strains expressing T7-PHO4, encoding Pho4p tagged with the T7 epitope, indicated that Pho2p interacts with T7-Pho4p and enhances the binding affinity of T7-Pho4p to the UAS. Protein binding experiments also showed that both Pho4p and Pho2p could bind with the general transcription factors, TBP, TFIIB, and TFIIEbeta, suggesting that the Pho4p-Pho2p complex bound to the UAS activates transcription of the PHO genes by direct interaction with the general transcription factors. PMID- 9354397 TI - Membranes of retinal microsomes: a new protein of the microsomal monooxigenase system. AB - A new component, which substitutes cytochrome P-450 as an acceptor of reducing equivalents from NADPH-cytochrome P-450 reductase, was identified in the bovine retina microsomal monooxigenase system, which does not contain cytochrome P-450. This component is a non-heme iron-containing protein with molecular mass of 66 kDa. The properties of the protein from the bovine retina are similar to those of MIP, a non-heme iron-containing protein from the heart microsomal monooxigenase system, in which cytochrome P-450 was not identified, either. Activation of the microsomal monooxigenase system (an increase in the NADPH-cytochrome P-450 reductase activity, an increased rate of microsomal NADPH oxidation) was shown in the retina upon long-term intensive illumination. It was shown also that the development of hereditary degeneration of the retina in rats was accompanied by activation of the specific microsomal monooxigenase system in the target tissues (retina, brain cortex) irrespective of its composition (cytochrome P-450 or non heme iron-containing protein). PMID- 9354396 TI - Extraction of functionally active photosystem 2 pigment-protein complexes from pea thylakoids and their purification on Sepharose DEAE 6B. AB - A method for selective extraction of photosystem 2 (PS 2) pigment-protein complexes (PPC) from pea thylakoids and their purification from Sepharose DEAE 6B was developed. It was shown that extraction of thylakoids (3 mg chlorophyll/ml) with Triton X-100 (Triton X-100:chlorophyll = 15-20:1 w/w) in the presence of 1 M sucrose, 1 M NaCl, 50 mM MES in the solubilization medium (pH 5.0), tenfold dilution with 50 mM MES buffer (pH 5.0), and centrifugation at 16,000 g for 10 min provided selective extraction of PS 2 PPC. Spectral, electrophoretic and functional properties of the isolated PPC were determined. This method allowed us to isolate native pigment-protein oxygen-evolving complexes (core complexes) and D1-D2-cytochrome b559 complexes of the reaction centre (RC). The core complex immobilized on a Sepharose DEAE 6B column was treated with 2 mM hydroxylamine or 1 M CaCl2. In the former case, modification by hydroxylamine led to the removal of 4 atoms of Mn, but not of the 33-kDa protein; in the latter case, modification by CaCl2 led to the removal of the 33 kDa protein. The modified core complexes were unstable upon native electrophoresis in the presence of n-dodecyl-beta-d maltoside and deriphat-160. We suggest that the structural organization of the Mn cluster and 33 kDa protein stabilized the core complex and increased its stability upon the action of the detergents. PMID- 9354398 TI - The effect of heat shock on M-cholinoreceptors from rat cerebral cortex membranes. AB - The major parameters of binding the specific blocker [3H]quinuclidinyl benzylate were obtained for M-cholinoreceptors of rat cerebral cortex membranes. One receptor was shown to bind two molecules of the ligand; two discrete receptor pools were revealed, which differed in the sensitivity (Kd1 = 0.35 +/- 0.03; Kd2 = 1.79 +/- 0.21 nM) and the number of the ligand binding sites (B1 = 468 +/- 54; B2 = 864 +/- 81 fmol/mg protein). Heat shock decreased the number of receptors of the high-affinity pool (B1) by 16% (Kd1 did not differ from the control); the efficiency of the ligand binding to the receptors (E = B/2Kd) decreased by 26%, which suggests the decrease in the functional activity of the receptors of this pool. In the low-affinity pool (B2), the number of receptors increased by 39%, and Kd2 decreased by 22%; the efficiency (E2) increased by 14%. PMID- 9354399 TI - Quantitative analysis of the movements of cytoplasmic granules in polarized fibroblasts. AB - Movements of cytoplasmic organelles were analyzed in Vero fibroblasts. In the cells polarized at the edge of an experimental wound, cytoplasmic granules moved randomly (Brownian motions) and by separate jumps (saltatory movements). The displacement of granules by the Brownian motions exceeded by more than an order of magnitude that of the mitochondria similar by weight. Lipid droplets moved predominantly by saltations, whereas mitochondria and lysosomes moved much less often. In a front part of the polarized cells, the main directions of saltatory movements were from the nucleus to the leading edge of a cell and back, whereas the tangential movements (across the long axis of a cell) were less than 1%. 90% of saltatory movements occurred in the area starting 10-12 microm from the nucleus and ending 10-12 microm from the leading edge of a cell. The average rate of saltatory movements of the granules (2.38 microm/s) was identical in both directions. The average length of the track was 7.49 microm; the maximum track length reached 30 microm. An increase in the granule diameter from 0.3 to 1.4 microm resulted in a minor (statistically insignificant) decrease in the average rate of the movements. The average rate of saltatory movements of mitochondria was 1.00 microm/s, and the average track length was 6.04 microm. Therefore, mitochondria, in contrast to lipid droplets, are rigidly fixed in the cytoplasm, and the force holding mitochondria is equal to the force produced by the microtubule-associated motors. Taking into account the characteristic of the centrifugal saltations, we suggest that they are mediated by an unusual dynein. PMID- 9354400 TI - The kinetic characteristics of L-type calcium channels in cardiac cells of hibernators. 2. Analysis of the kinetic model. AB - The present paper describes the experimental and theoretical investigations of the kinetic characteristics of the L-type Ca2+ channels in ground squirrels Citellus undulatus in two different physiological states (hibernation and spontaneous arousal). The perforated patch-clamp method was used in the experiments. We have shown in the previous study [1] that Ca2+ currents in hibernating and active animals may be described by the d2f1(2)f2 model. Based on that model, in this paper we studied in detail the main steps of the conductance regulation of Ca2+ channels: activation (d), slow (f1-type) and fast (f2-type) inactivations of the channel. Activation is related to the movement of the gating charge. Slow inactivation is associated with the movement of the gating charge and is current-dependent. Fast inactivation is a more complex process and cannot be represented as a single-stage conformational transition induced by the gating charge movement. It is regulated by cAMP phosphorylation. The differences in the Ca2+ current kinetics are observed virtually for all the components. In hibernating animals, the most pronounced shift (15-20 mV) towards depolarization is experienced by the normalized conductances of both inactivation components, whereas the conductance of the activation component is shifted to a lesser extent. The characteristic times of Ca2+ currents of hibernating ground squirrels are 1.5-2 times greater than those of aroused animals. The current activation gating charge of Ca2+ channels in ground squirrel cardiocytes was found to change. The gating charge was about 2 in hibernating animals and 1.5 in active squirrels. The effect of isoproterenol-induced cAMP-dependent phosphorylation on Ca2+ currents in cardiocytes from hibernating ground squirrels was studied. Isoproterenol restored the kinetic parameters of Ca2+ currents to the values close to the parameters of active animals. However, we failed to explain the suppression of the Ca2+ current in hibernating animals in terms of cAMP-dependent regulation only. PMID- 9354401 TI - The effect of arachidonic acid on junctional conductance in isolated murine hepatocytes. AB - Arachidonic acid (AA) is known to inhibit intercellular conductance in normal and tumour cells. We showed that junctional conductance (Gj) in isolated murine hepatocytes was relatively tolerant to the uncoupling effect of AA. Extracellular application of 100 microM AA decreased Gj in less than 50% of hepatocytes, and the effect was much slower in other cells (10-15 min vs. 2-5 min, respectively). The uncoupling effect of AA did not depend on the intracellular [Ca2+] within the pCa(i) range 7.7-9.0. Similar results were obtained using the pipette-filling solutions with low (1 mM) and high (10 mM) concentrations of a Ca-chelating agent (EGTA). To verify whether the resistance of the hepatocyte Gj to AA may result from the "wash-out" of the intracellular intermediates during the intracellular dialysis, Gj was measured 10-45 min after the preincubation of hepatocytes with AA. After such a treatment, in 62% of cell pairs the Gj values recorded did not differ from the control. Extracellular or intracellular acidification (pHo 6.0 or pHi 5.0-6.0) did not markedly affect the AA action. However, in some cases AA induced the recovery of Gj blocked after intracellular acidification, the phenomenon suggesting the activation of the H+ transport in the presence of AA. Possible mechanisms of the observed resistance of junctional conductance of mouse hepatocytes in primary culture are discussed. PMID- 9354402 TI - Properties of the biomembrane lipid phase optimization to high pressure. AB - The hypothesis of homeoviscous adaptation extended to other (except viscosity) physical properties of membranes raises the problem of the regulation of several membrane properties under environmental changes. The existence of limitations on the lipid composition, required for maintaining membrane polarity and viscosity at constant levels, was demonstrated for model membranes (liposomes from egg yolk lecithin and cholesterol) at the varying hydrostatic pressure (0-600 atm). It is assumed that similar limitations can determine the features of the lipid composition of biological membranes. PMID- 9354403 TI - Effect of sodium chloride, chlorite, and perchlorate on the hypochlorite-induced peroxidation of phospholipid liposomes. AB - The abilities of sodium hypochlorite (NaClO), chlorite (NaClO2), chlorate (NaClO3), and perchlorate (NaClO4) to initiate lipid peroxidation (LP) in liposomes formed from unsaturated phosphatidylcholine were compared. It was shown that only NaClO induced an intensive accumulation of LP products (thiobarbituric acid-reactive substances and diene conjugates) in the liposomes as a result of their co-incubation. The other oxochlorates produced no similar effects and did not affect the hypochlorite-induced LP. This indicates that the observed hypochlorite-induced LP does not result from the presence of chlorite, chlorate, or perchlorate anion admixtures in the medium. PMID- 9354404 TI - A mathematical model of T lymphocyte proliferation controlled by interleukin-2 internalization. AB - A mathematical model of the first in vitro IL-2-controlled division cycle of T lymphocytes is presented. The model describes the population dynamics of T cells at different stages of the cell cycle and introduces "molecular" equations for the G1-S phase control. These equations are based on the current knowledge of the biochemical mechanisms of ligand-receptor binding, ligand and receptor synthesis, and internalization of the ligand-receptor complexes. The temporal order in the biosynthesis of IL-2 and IL-2R during the G1a and G1b phases is investigated. We show numerically that the maximum number of cells in the S-phase depends on the reversibility of the G1a-G1b transitions, on the rates of ligand-receptor binding, ligand and receptor synthesis, and internalization of the ligand receptor complexes. The phase portraits for different hypotheses on the temporal order of the IL-2 and high affinity receptor synthesis at the G1 phase of the cell cycle are qualitatively different. On this basis, simple kinetic experiments to distinguish between these alternatives are proposed. PMID- 9354405 TI - An operational amplifier B1404UD1A-1 in the patch-clamp current-to-voltage converter. AB - The applicability of the home-made operational amplifier B1404UD1A-1 in a patch clamp current-to-voltage converter was analyzed. Its parameters (background noise, input bias current, and gain-bandwidth product) were estimated. Schematic solutions and practical recommendations for the use of this amplifier in a current-to-voltage converter were given. Based on the background noise and frequency parameters of the converter, we found that this device can be used for measuring ion channel currents with a high sensitivity and within a broad frequency range (0.055 pA, to 1 kHz; 0.4 pA, to 10 kHz). An example of the converter application in experiments is given. PMID- 9354406 TI - Introduction: changing trends in antiarrhythmic therapy: worldwide experience with class III agents. PMID- 9354407 TI - Controlling cardiac arrhythmias: an overview with a historical perspective. AB - During the past decade, several developments in our knowledge of antiarrhythmic drugs have had a major influence on our approach to their use. These developments may be summarized as follows: (1) it has become clear that arrhythmias merit treatment only for the relief of symptoms, with improved quality of life, and for prolongation of survival by reducing arrhythmic deaths; (2) suppression of arrhythmias--symptomatic or asymptomatic--may not necessarily decrease mortality, the net impact on mortality being agent-specific; (3) antiarrhythmic drugs have the propensity to decrease as well as to increase cardiac arrhythmias (producing proarrhythmias); (4) the most important determinant of arrhythmia mortality is the degree and nature of ventricular dysfunction; and (5) only controlled trials have the potential to establish the effect of treatment on mortality in patients with cardiac arrhythmias. To these considerations must be added the advances in nonpharmacologic approaches to controlling cardiac arrhythmias. These include catheter ablation of cardiac arrhythmias, certain surgical techniques that in selected patients offer prospects of cure, and the development of implantable ventricular and atrial cardioverter defibrillators, which allow the evaluation of drugs versus placebo against the background of the defibrillator. This is particularly germane in the case of life-threatening symptomatic ventricular arrhythmias such as sustained ventricular tachycardia and ventricular fibrillation. Antiarrhythmic drugs and implantable devices in the control of arrhythmias cannot be considered in isolation. Their role in mortality reduction needs to be defined alone as well as in combination by controlled clinical trials. PMID- 9354408 TI - Pharmacologic and pharmacokinetic profile of class III antiarrhythmic drugs. AB - Cardiac arrhythmias frequently respond only to drugs that have as their predominant electrophysiologic effect the prolongation of repolarization and refractoriness. According to the Singh-Vaughan Williams classification, these drugs are known as class III agents. In the last few years, interest has increased in the development of class III antiarrhythmic drugs as alternatives to sodium channel blocking agents, which mainly affect cardiac conduction. Much of this interest results from a perceived danger of using drugs with sodium channel blocking properties, particularly in patients with ischemic heart disease, based on the results of the Cardiac Arrhythmia Suppression Trial (CAST) and several other trials. This article is a review of the pharmacology, including the pharmacokinetics and pharmacodynamics, of the most commonly used and investigated class III antiarrhythmic drugs. As will be seen from the discussion, each of these drugs has novel pharmacology that makes it applicable in specific clinical situations. Their putative effects on various arrhythmogenic mechanisms and their efficacy in treating specific target arrhythmias will be addressed. PMID- 9354409 TI - Hemodynamic effects of antiarrhythmic compounds: intrinsic effects and autonomic modulation. AB - Besides their proarrhythmic side-effects, most antiarrhythmic drugs exert varying degrees of depressant action on hemodynamics, which may limit their utility, especially in patients with compromised left ventricular function. Antiarrhythmic drugs have not only myocardial inotropic effects but also act on the coronary and peripheral circulation and the heart rate. Thus, sophisticated and appropriate experimental conditions are necessary to define the effect of their direct negative inotropic actions versus their circulatory effects and the impact of drug-induced autonomic modulation. This study describes an extended comparison of amiodarone, d-sotalol, d,l-sotalol, and dofetilide as class III antiarrhythmic drugs with the actions of several class I antiarrhythmic drugs in an open-chest model. The experimental model permits not only measurements in the intact circulation but also measurements of isovolumic indexes of contractility, which are independent of drug-induced changes in ventricular preload and afterload. Furthermore, after autonomic blockade, hemodynamic effects can be measured independently of modulatory adrenergic effects in such a model. d-Sotalol and amiodarone had cardiodepressant effects only at doses significantly higher than the highest doses used clinically. Dofetilide did not have a negative inotropic effect at doses up to 40 ng/kg. However, these results might be modified in experimental models with severely compromised left ventricular function, as was shown for class I antiarrhythmic drugs and for d,l- and d-sotalol. The sensitivity to a drug's negative inotropic action is markedly increased in functionally impaired myocardium. Furthermore, in a model of postischemic myocardial dysfunction, the depressant effect of d-sotalol could largely be avoided by previous autonomic blockade, indicating the importance of the residual beta-blocking potency of d-sotalol in the doses used in our experiments. Thus, in clinically relevant doses amiodarone, d-sotalol, and dofetilide were found to be devoid of negative inotropic actions in the setting of normal left ventricular myocardium. In failing hearts, such effects become more readily evident than they do in normal hearts after high doses of amiodarone and d-sotalol. From beta blocking experiments in hearts with left ventricular dysfunction, it could be inferred that residual beta-blocking and other negative inotropic mechanisms may produce a net negative inotropic action, thus masking the minor positive class III effects postulated from in vitro experiments. These observations may have significant clinical implications, because they suggest that the intrinsic myocardial effects of antiarrhythmic drugs may be modified by autonomic effects, the status of ventricular function, and changes in preload and afterload. PMID- 9354410 TI - General principles of antiarrhythmic therapy for ventricular tachyarrhythmias. AB - Sudden cardiac death due to ventricular arrhythmias is a significant cause of mortality in patients with structural heart disease. Over the past several decades, the introduction of new pharmacologic and nonpharmacologic therapy has expanded the treatment options available. This article will focus on the use of antiarrhythmic medication for the treatment of ventricular arrhythmias and will review the following: (1) treatment goals for various clinical populations, (2) the mechanisms of antiarrhythmic and proarrhythmic actions of antiarrhythmic medications, and (3) empiric versus guided pharmacologic therapy. PMID- 9354411 TI - Prolonging survival by reducing arrhythmic death: pharmacologic therapy of ventricular tachycardia and fibrillation. AB - Antiarrhythmic drug therapy of sustained ventricular tachyarrhythmias is undertaken to reduce arrhythmic symptoms, recurrences, and mortality. Ideally, reduction of arrhythmic death will reduce total mortality as well, although this may not hold true in the presence of competing risk. Whether, in fact, antiarrhythmic therapy actually reduces arrhythmic death remains uncertain in the absence of any placebo-controlled trials. Nonetheless, the following conclusions can be drawn from the Electrophysiologic Study versus Electrocardiographic Monitoring (ESVEM) trial, the Cardiac Arrest Study Hamburg (CASH), and the Cardiac Arrest in Seattle: Conventional versus Amiodarone Drug Evaluation (CASCADE) study, as well as a beta blocker study by Steinbeck et al: (1) class I antiarrhythmics are less effective than amiodarone or sotalol for the prevention of recurrent sustained ventricular tachycardia/ventricular fibrillation; (2) sympathetic inhibition as a component of the antiarrhythmic regimen may strongly contribute to mortality reduction; and (3) the respective roles of antiarrhythmic drugs, implantable devices, and the concurrent use of both are in a state of flux, awaiting results of randomized controlled clinical trials. PMID- 9354412 TI - Ablation therapy for cardiac arrhythmias. AB - Ablation has become an important and, in some cases, the first-line therapy for a number of tachyarrhythmias. The feasibility of treating arrhythmias with ablation was initially demonstrated with surgical ablation techniques. Recently, catheter ablation techniques have replaced the surgical approach in nearly all cases. Catheter ablation is highly effective for the Wolff-Parkinson-White syndrome, atrioventricular nodal reentry, and atrial ectopic tachycardia. It is effective for atrial flutter, although approximately one quarter of patients treated with catheter ablation continue to require therapy for concomitant atrial fibrillation. The surgical maze procedure has proved to be feasible for preventing atrial fibrillation. The risks and long-term efficacy of catheter ablation maze procedures for atrial fibrillation need to be defined. The efficacy of ablation for ventricular tachycardia varies with the type of tachycardia. Catheter ablation is very effective for the rare idiopathic ventricular tachycardias that occur in structurally normal hearts and for bundle-branch reentry ventricular tachycardia, which occurs most frequently in patients with dilated cardiomyopathy. When performed at an experienced center, surgical ablation is an excellent option for selected patients with ventricular tachycardia due to prior myocardial infarction who have a discrete aneurysm but otherwise well-preserved ventricular function. Catheter ablation shows promise for this arrhythmia, but it can be offered only to those patients who have relatively slow tachycardias that allow catheter mapping. Substantial advances in mapping and ablation technology will continue to occur, allowing nonpharmacologic control of cardiac arrhythmias to be achieved in an ever greater number of patients. PMID- 9354413 TI - Drugs versus devices in controlling ventricular tachycardia, ventricular fibrillation, and recurrent cardiac arrest. AB - Patients with symptomatic ventricular tachycardia, ventricular fibrillation, or aborted sudden cardiac death remain at high risk for arrhythmia recurrence. In recent years, strategies to treat these patients have changed. Concerns about the proarrhythmia risk and uncertain efficacy of class I agents have resulted in a shift in interest to non-class I antiarrhythmic drugs such as sotalol and amiodarone. Both drugs have class III antiarrhythmic properties (i.e., both lengthen repolarization and refractoriness); however, each also has its own additional electrophysiologic effects. Prospectively designed, randomized studies have shown that both sotalol and amiodarone have more potent antiarrhythmic actions than class I agents. However, even as the advantages of sotalol and amiodarone have been recognized, enthusiasm for nonpharmacologic modes of treatment, particularly the implantable cardioverter-defibrillator (ICD), has also markedly increased. The ICD has been shown to decrease dramatically the incidence of sudden death, which may lead to the reduction of total mortality. Whether patients with life-threatening ventricular tachyarrhythmias should be treated first with antiarrhythmic agents or with an ICD is an important question. The results of recent studies suggest that treatment with an ICD is more effective than electrophysiologically guided treatment with class I agents. However, results of prospectively designed randomized studies comparing the efficacy of the ICD with that of sotalol and amiodarone must become available before definitive recommendations can be made concerning the use of the ICD as first-line therapy in patients with ventricular tachycardia/ventricular fibrillation or aborted sudden cardiac death. In addition, there may be a significant role for the use of antiarrhythmic drugs in conjunction with ICDs. PMID- 9354414 TI - Atrial fibrillation: antiarrhythmic therapy versus rate control with antithrombotic therapy. AB - Atrial fibrillation is a major health problem in the United States, but the best strategies for treating it have not been rigorously determined in clinical studies. Specifically, there is a paucity of data comparing the approach of maintaining sinus rhythm using prophylactic antiarrhythmic drug therapy with the approach of controlling the ventricular response to atrial fibrillation while reducing embolic events with concomitant antithrombotic therapy. Until ongoing randomized trials are completed, which patients benefit most from a specific approach cannot be determined with certainty. In general, the most reasonable strategies include (1) the restoration of sinus rhythm (without prophylactic antiarrhythmic therapy) after the patient's first episode of atrial fibrillation; and (2) the maintenance of sinus rhythm (including the use of prophylactic antiarrhythmic therapy) in patients who remain symptomatic despite adequate rate control, and who are not at high risk for proarrhythmia and/or are unlikely to maintain sinus rhythm. The risks and benefits need to be carefully weighed in patients with truly asymptomatic atrial fibrillation. Many patients may require multiple attempts to maintain sinus rhythm. Current investigative treatment modalities (e.g., ablation techniques, atrial implantable cardioverter defibrillators, new antiarrhythmic agents) are likely to alter the current approaches to atrial fibrillation. PMID- 9354415 TI - Proarrhythmia with class III antiarrhythmic drugs: types, risks, and management. AB - The nature of the proarrhythmic reactions induced by antiarrhythmic drugs is linked to the electrophysiologic effects of these agents. Torsades de pointes is the classic form of proarrhythmia observed during therapy with any drug that prolongs repolarization, for example, the class III agents. Its precise electrophysiologic mechanism is not fully elucidated, although the arrhythmia is generally considered to be due either to early afterdepolarization in the context of prolonged cardiac repolarization or to an increase in spatial or temporal dispersion of repolarization. Among the class III drugs the proarrhythmic risk appears to be lowest for amiodarone, probably due to its complex electrophysiologic profile that may create significant myocardial electrical homogeneity. In the case of d,l-sotalol, the incidence of torsades de pointes increases with dose and the baseline values of the QT interval. Where d-sotalol and other pure class III agents might fall into the varying spectrum of proarrhythmic potential remains unclear. That d-sotalol has been found to increase mortality in postinfarction patients with ventricular dysfunction (the Survival With Oral d-Sotalol [SWORD] trial) is a matter of considerable concern. It raises the possibility that such a phenomenon may be a common property of most, if not all, pure class III compounds. Accordingly, care must be taken to minimize the likelihood of proarrhythmia; in particular, therapy with a class III agent should only be initiated in the presence of a defined indication established on the basis of clinical trials. When class III antiarrhythmic drug induced proarrhythmia occurs, immediate cessation of therapy with the responsible agent and correction of predisposing factors, such as electrolyte disorders or bradycardia, is mandatory. Intravenous administration of high-dose magnesium sulfate has been demonstrated to be effective in terminating and preventing new episodes of torsades de pointes. Temporary pacing may be necessary. PMID- 9354416 TI - The side effect profile of class III antiarrhythmic drugs: focus on d,l-sotalol. AB - Class III antiarrhythmic drugs have been under extensive clinical investigation as safer, more effective alternatives to class I drugs, which have recognized risks in selected populations. Class III drugs prolong the action potential duration of myocardial cells, resulting in a lengthening of the effective refractory period. This pharmacologic activity has antiarrhythmic properties, but it may induce a distinctive form of proarrhythmia known as torsades de pointes. Amiodarone and d,l-sotolol are class III drugs that have been available for many years. In addition to their ability to prolong refractoriness, these drugs have other pharmacodynamic properties. Recent antiarrhythmic drug discovery has focused on the identification and development of selective or so-called pure class III drugs that are devoid of additional actions. Investigators have hoped that these drugs would be as effective as sotalol and amiodarone but have fewer adverse effects. Accumulating data, however, indicate that complex compounds exhibiting antiadrenergic and other electrophysiologic properties may be superior to pure class III agents. PMID- 9354417 TI - Antiarrhythmic therapy--future trends and forecast for the 21st century. AB - This article discusses recent changes in antiarrhythmic therapy, with a focus on nonpharmacologic therapy (electrode catheter ablation, implantable cardioverter defibrillators [ICDs]), and puts them into perspective for the coming years. The treatment of supraventricular tachycardias and tachycardia involving accessory pathways is likely to remain the domain of catheter ablation. With promising new techniques under investigation, the spectrum of arrhythmias that can be cured will probably be expanded. Treatment of life-threatening ventricular arrhythmias is likely to remain the domain of the ICD in the foreseeable future. With the safety net of the ICD in place, new antiarrhythmic drugs or other forms of antiarrhythmic therapy can be developed and tested. PMID- 9354418 TI - Dietary restriction reduces insulin-like growth factor I levels, which modulates apoptosis, cell proliferation, and tumor progression in p53-deficient mice. AB - Diet contributes to over one-third of cancer deaths in the Western world, yet the factors in the diet that influence cancer are not elucidated. A reduction in caloric intake dramatically slows cancer progression in rodents, and this may be a major contribution to dietary effects on cancer. Insulin-like growth factor I (IGF-I) is lowered during dietary restriction (DR) in both humans and rats. Because IGF-I modulates cell proliferation, apoptosis, and tumorigenesis, the mechanisms behind the protective effects of DR may depend on the reduction of this multifaceted growth factor. To test this hypothesis, IGF-I was restored during DR to ascertain if lowering of IGF-I was central to slowing bladder cancer progression during DR. Heterozygous p53-deficient mice received a bladder carcinogen, p-cresidine, to induce preneoplasia. After confirmation of bladder urothelial preneoplasia, the mice were divided into three groups: (a) ad libitum; (b) 20% DR; and (c) 20% DR plus IGF-I (IGF-I/DR). Serum IGF-I was lowered 24% by DR but was completely restored in the IGF-I/DR-treated mice using recombinant IGF I administered via osmotic minipumps. Although tumor progression was decreased by DR, restoration of IGF-I serum levels in DR-treated mice increased the stage of the cancers. Furthermore, IGF-I modulated tumor progression independent of changes in body weight. Rates of apoptosis in the preneoplastic lesions were 10 times higher in DR-treated mice compared to those in IGF/DR- and ad libitum treated mice. Administration of IGF-I to DR-treated mice also stimulated cell proliferation 6-fold in hyperplastic foci. In conclusion, DR lowered IGF-I levels, thereby favoring apoptosis over cell proliferation and ultimately slowing tumor progression. This is the first mechanistic study demonstrating that IGF-I supplementation abrogates the protective effect of DR on neoplastic progression. PMID- 9354419 TI - Hyperlipidemia enhancement of renal preneoplasia but not tumor formation is due to elevated apoptosis in the Eker rat. AB - The influence of a high-fat diet on the appearance of renal tumors was assessed in the Eker rat model of hereditary renal carcinoma. Examination of H&E-stained sections showed a significant increase in the number of microscopic solid adenomas in the high-fat group compared with the low-fat group, whereas there was no significant difference in the number of macroscopic tumors between the two groups. Where had the tumor buds gone? Staining for apoptotic bodies occasionally revealed apoptosis in and around the microscopic adenomas. In addition, an Eker rat renal tumor-derived cell line showed apoptosis when it was cultured with high concentrations of native and acetylated low-density lipoprotein. These findings suggested that tumor buds repeatedly appeared and disappeared in Eker rats on a high-fat diet. PMID- 9354420 TI - Trichoepitheliomas contain somatic mutations in the overexpressed PTCH gene: support for a gatekeeper mechanism in skin tumorigenesis. AB - The nevoid basal cell carcinoma (Gorlin) syndrome (NBCCS) is an autosomal dominant disorder characterized by multiple developmental defects and cancer susceptibility. NBCCS is caused by mutations in the human homologue (PTCH) of the Drosophila patched gene, a developmental regulator implicated in signaling of hedgehog and smoothened. The PTCH gene was found to contain somatic mutations also in sporadic basal cell carcinomas and medulloblastomas, tumors seen in NBCCS, consistent with PTCH acting as a tumor suppressor. Because basal cell carcinomas have been observed to develop in association with benign trichoepitheliomas (TEs) in the same lesions, patients, and families and may share the same cell of origin, we have analyzed PTCH for mutations and expression in TEs. We report frameshift and in-frame somatic deletions in this gene and a consistent overexpression of PTCH mRNA in TEs. These findings provide the first evidence of a gene mutation in TEs and identify a common pathogenic pathway for histopathologically similar but prognostically distinct skin tumors. Moreover, these results support the presence of a gatekeeper mechanism in multistep skin tumorigenesis exerted by the altered PTCH signaling pathway. PMID- 9354421 TI - Somatic mutations of the MEN1 tumor suppressor gene in sporadic gastrinomas and insulinomas. AB - Gastrinomas and insulinomas are frequent in multiple endocrine neoplasia type 1 (MEN1). The MEN1 tumor suppressor gene was recently identified. To elucidate the etiological role of the MEN1 gene in sporadic enteropancreatic endocrine tumorigenesis, we analyzed tumors (28 gastrinomas and 12 insulinomas) from 40 patients for MEN1 gene mutations and allelic deletions. One copy of the MEN1 gene was found to be deleted in 25 of 27 (93%) sporadic gastrinomas and in 6 of 12 (50%) sporadic insulinomas. MEN1 gene mutations were identified in 9 of 27 (33%) sporadic gastrinomas and 2 of 12 (17%) insulinomas and were not seen in corresponding germ-line DNA sequence. A specific MEN1 mutation was detected in one gastrinoma and in the corresponding germ-line DNA of a patient who had no family history of MEN1. Somatic MEN1 gene mutations and deletions play a critical role in the tumorigenesis of sporadic gastrinomas and may also contribute to the development of a subgroup of insulinomas. PMID- 9354422 TI - Androgen-independent prostate cancer progression in the TRAMP model. AB - We previously established the autochthonous transgenic adenocarcinoma mouse prostate (TRAMP) model to facilitate characterization of molecular mechanisms involved in the initiation and progression of prostate cancer. TRAMP mice display high grade prostatic intraepithelial neoplasia or well-differentiated prostate cancer by 10-12 weeks of age. To test the hypothesis that molecular events leading to androgen independence and metastasis can occur early in the natural history of prostate cancer yet remain silent until selective pressures such as androgen deprivation are applied, we have examined the consequences of castration on the initiation and progression to metastatic prostate cancer in TRAMP mice. Cohorts were castrated at 12 weeks of age and sacrificed at 18 (T12/18) or 24 (T12/24) weeks of age, and the development of primary cancer and metastatic disease was compared to noncastrated (T18 and T24) controls. Median T12/18 and T12/24 genitourinary (GU) weight was significantly less than T18 and T24, respectively. In addition, T12/24 GU weight was significantly greater than T12/18. Histological prostate tumors developed in 3 of 7 T12/18 and 8 of 10 T12/24 mice. All tumors that developed in castrated mice were poorly differentiated in contrast to 27% in noncastrated controls. Although castration significantly decreased GU tumor burden, overall progression to poorly differentiated and metastatic disease was not ultimately delayed. These results demonstrate that prostate cancer in the TRAMP model is heterogeneous with respect to androgen dependence as early as 12 weeks of age; therefore, early androgen ablation may have a variable impact on progression in an individual mouse. Further analysis of this prostate cancer model to identify specific molecular mechanisms that determine androgen sensitivity may facilitate future initiation of appropriate individualized hormonal therapy for the management of human prostate cancer. PMID- 9354423 TI - Loss of FHIT expression in cervical carcinoma cell lines and primary tumors. AB - Allelic deletions involving the short arm of chromosome 3 (3p13-21.1) have been observed frequently in cervical carcinomas. Recently, a candidate tumor suppressor gene, FHIT (Fragile Histidine Triad), was cloned and mapped to this chromosomal region (3p14.2). Abnormal FHIT transcripts have been identified previously in a variety of tumor cell lines and primary carcinomas, although their significance and the molecular mechanisms underlying their origin remain incompletely defined. In addition, integration of human papillomavirus DNA has been identified at a fragile site (FRA3B) within the FHIT locus in cervical cancer. These observations motivated us to evaluate FHIT mRNA and protein expression in cervical cancer cell lines, primary cervical carcinomas, and normal tissues. Transcripts of the expected size and sequence were the predominant species identified by reverse transcription (RT)-PCR in cultured keratinocytes and all normal tissues evaluated. In contrast, aberrant FHIT transcripts were readily demonstrated in 6 of 7 cervical carcinoma cell lines and 17 of 25 (68%) primary cervical carcinomas. Northern blot analyses demonstrated reduced or absent FHIT expression in the cervical carcinoma cell lines, particularly those with aberrant RT-PCR products. Immunohistochemical analysis of Fhit expression in cervical tissues revealed strong immunoreactivity in nonneoplastic squamous and glandular cervical epithelium and marked reduction or loss of Fhit protein in 25 of 33 (76%) primary cervical carcinomas. In those cervical cancer cell lines and primary tumors with exclusively aberrant or absent FHIT transcripts by RT-PCR, Fhit protein expression was always markedly reduced or absent. The frequent alterations in FHIT expression in many cervical carcinomas, but not in normal tissues, suggest that FHIT gene alterations may play an important role in cervical tumorigenesis. PMID- 9354424 TI - A precise interchromosomal reciprocal exchange between hot spots for cleavable complex formation by topoisomerase II in amsacrine-treated Chinese hamster ovary cells. AB - Among the aprt mutations induced in confluence-arrested Chinese hamster ovary D422 cells by the topoisomerase II poison amsacrine, there was a reciprocal exchange between the aprt gene and an unrelated sequence, accompanied by a chromosomal translocation at the aprt locus. The breakpoints in both parental sequences were hot spots for amsacrine-stimulated DNA cleavage in vitro, and the novel junctions formed were precisely as expected for a mechanism involving reciprocal exchange of topoisomerase II subunits followed by resealing of the breaks and correction of mismatches in the cohesive ends. The results are consistent with a role for direct subunit exchange in the production of chromosomal translocations by topoisomerase poisons, although more complex models involving double-strand breakage and repair could produce reciprocal exchanges of similar specificity. PMID- 9354425 TI - Loss of p21Waf1/Cip1 sensitizes tumors to radiation by an apoptosis-independent mechanism. AB - Cellular checkpoints are important mediators of the response of normal cells following genotoxic damage, and interruption of these checkpoints is a common feature of many solid tumors. Although the effects of loss in checkpoint function in tumor cells are well understood in terms of cell cycle control, there is little information on their role in determining treatment efficacy in vivo. We have examined both the in vitro and in vivo responses of isogenic lines differing only in the p53-transactivated checkpoint gene, p21Waf1/Cip1. When assayed in vitro, loss of p21 in human colon tumor cells results in a selective induction of apoptosis [Waldman, T., et al., Nature (Lond.), 381: 713-716, 1996.] but no difference in the clonogenic survival. However, when grown as xenografts and irradiated in situ, p21-deficient tumors were significantly more sensitive to radiation as assessed both by clonogenic survival and by regrowth of the tumors following treatment. These data indicate that loss of p21 results in increased sensitivity to killing by ionizing radiation that is independent of the induction of apoptosis and cell cycle arrest but that is specific to cells when they are grown as a solid tumor. These results have important implications for assessing both the genetic determinants of sensitivity to anticancer agents and efficacy of anticancer agents. PMID- 9354426 TI - Early age at diagnosis in families providing evidence of linkage to the hereditary prostate cancer locus (HPC1) on chromosome 1. AB - In a recent study of 91 families having at least three first degree relatives with prostate cancer, we reported the localization of a major susceptibility locus for prostate cancer (HPC1) to chromosome 1 [band q24; J. R. Smith et al., Science (Washington DC), 274: 1371-1373, 1996]. There was significant evidence for locus heterogeneity, with an estimate of 34% of the families being linked to this locus. In this report, we investigate the importance of age at diagnosis of prostate cancer and number of affected individuals within a family as variables in the linkage analysis of an expanded set of markers on 1q24. Under two different models for the prostate cancer locus, we find that the evidence for linkage to HPC1 is provided primarily by large (five or more members affected) families with an early average age at diagnosis. Specifically, for 40 North American families with an average age at diagnosis <65 years, the multipoint lod score is 3.96, whereas for 39 families with an older average age at diagnosis, this value is -0.84. Assuming heterogeneity, the proportion of families linked is 66% for the 14 families with the earliest average ages at diagnoses, but it decreases to 7% for the families with the latest ages at diagnoses. A similar age effect is observed in 12 Swedish pedigrees analyzed. To test the hypotheses generated by these analyses, we examined an additional group of 13 newly identified prostate cancer families. Overall, these families provided additional evidence for linkage to this region (nonparametric linkage Z = 1.91; P = 0.04 at marker D1S1660), contributed primarily by the families in this group with early age at diagnosis [nonparametric linkage Z = 2.50 (P = 0.01) at D1S422]. These results are consistent with the existence of a locus in this region that predisposes men to develop early-onset prostate cancer. PMID- 9354427 TI - Somatic deletions and mutations in the Cowden disease gene, PTEN, in sporadic thyroid tumors. AB - The majority of familial medullary thyroid neoplasms are associated with germ line mutations of the RET proto-oncogene, yet very little is known about the mechanisms involved in the pathogenesis of familial and sporadic nonmedullary thyroid tumors. A subset of thyroid tumors have loss of heterozygosity of chromosome 10q22-23, a region harboring the gene responsible for Cowden disease, an autosomal dominant hamartoma syndrome associated with thyroid and breast tumors. PTEN/MMAC1/TEP1 codes for a dual-specificity phosphatase and is likely a tumor suppressor gene. We sought to determine the PTEN status in a series of epithelial thyroid neoplasms. We studied 95 sporadic thyroid tumors, of which 39 were papillary thyroid carcinomas (PTCs), 12 were follicular carcinomas, 9 were anaplastic carcinomas, 5 were Hurthle cell carcinomas, 21 were nonfunctioning follicular adenomas, and 9 were Hurthle cell adenomas. Direct sequencing of PCR amplified products was performed for all nine exons of PTEN. Two polymorphic markers, one located in intron 8 and another, a dinucleotide repeat marker, AFMa086wg9, located within intron 2, were analyzed in paired blood-tumor DNA samples to assess hemizygous deletions of PTEN. We found a somatic frameshift mutation in one PTC, which was expected to generate a premature stop codon 2 amino acids downstream. Twenty-six % of informative benign tumors (four follicular adenomas and three Hurthle cell adenomas) and only 3 of 49 (6.1%) informative malignant tumors (one PTC, one follicular carcinoma, and one anaplastic carcinoma) showed evidence of hemizygous deletion of PTEN (P = 0.046). We conclude that a subset of thyroid tumors have somatic deletions of the PTEN gene, predominantly the benign forms, and that small intragenic mutations of PTEN are infrequent in thyroid tumors. We speculate that other mechanisms of PTEN inactivation, rather than small intragenic mutations, might occur in the hemizygously deleted samples and act as the "Knudson second hit." Alternatively, other tumor suppressor genes mapping to chromosome 10q22-23 could be the actual targets for such deletions and thus represent the various hits in the pathway of multistep carcinogenesis. PMID- 9354428 TI - Activated Ki-ras enhances sensitivity of ceramide-induced apoptosis without c-Jun NH2-terminal kinase/stress-activated protein kinase or extracellular signal regulated kinase activation in human colon cancer cells. AB - The activation of c-Jun NH2-terminal kinase (JNK)/stress-activated protein kinase (SAPK) and/or extracellular signal-regulated kinase (ERK) is involved in ceramide induced apoptosis in certain cells. To examine the relationship between activated Ki-ras-mediated signals and ceramide-induced apoptosis in human colon cancer cells, JNK/SAPK and ERK activity, as initiated by ceramide, was examined in HCT116, which has a mutation of Ki-ras at codon 13, and HCT116-derived clones, HKe-3 and HKh-2, in which activated Ki-ras was disrupted through gene targeting. In HKe-3 and HKh-2, the activity of JNK/SAPK increased significantly within 60 min following C2 ceramide stimulation, and some apoptosis followed. In contrast, C2 ceramide caused a marked apoptosis in HCT116, but activation of JNK/SAPK was not observed. C2 ceramide did not activate ERK in any of the cell lines. These results suggest that activated Ki-ras contributes to the sensitivity of ceramide induced apoptosis without JNK/SAPK or ERK activation and that other signaling pathways involved in ceramide-induced apoptosis may be present in human colon cancer cells. PMID- 9354429 TI - Association of human T-cell leukemia virus and myelodysplastic syndrome in a central European population. AB - In addition to a few disorders such as acute T-cell leukemia that are typically associated with the human T-cell leukemia virus (HTLV) 1 in endemic regions, this virus may also play a role in some other hematological diseases. Here, we examine the incidence of HTLV in hematological diseases from a nonendemic region in central Europe. Data obtained by PCR and/or serological techniques from a total of 730 cases showed that besides the expected presence of HTLV-1 in T-lymphoid diseases (2 of 27 cases), HTLV-1 was only detected in myelodysplastic syndrome (MDS), in which an incidence of 17% (11 of 65 cases) was found. A correlation with a history of multiple transfusions or treatment with blood products in the HTLV-1-positive MDS could not be ascertained. Cytogenetics detected the presence of del(5)(q) in six HTLV-positive cases (five MDS and one T-cell acute lymphocytic leukemia) but in only one HTLV-negative case. These data indicate that allelic deletions of a series of 5q-located genes that typically occur in MDS may be associated with HTLV infections in central Europe. PMID- 9354431 TI - Sunlight induces pyrimidine dimers preferentially at 5-methylcytosine bases. AB - The most prevalent DNA lesion induced by UV irradiation is the cyclobutane pyrimidine dimer (CPD), which forms at positions of neighboring pyrimidines. Here we show that the rare DNA base 5-methylcytosine is the preferred target for CPD formation when cells are irradiated with natural sunlight. We have mapped the distribution of CPDs formed in normal human keratinocytes along exons of the p53 gene. Codons 196, 245, 248, and 282, which are mutational hot spots in skin cancers, are only weakly to moderately susceptible to formation of CPDs after irradiation with UVC (254 nm) or UVB (320 nm) light sources. However, when cells were exposed to natural sunlight, CPD formation was enhanced up to 15-fold at these codons due to the presence of 5-methylcytosine bases. These results suggest that CPDs containing 5-methylcytosine may play an important role in formation of sunlight-induced skin tumors and that methylation of CpG sequences, besides being involved in spontaneous mutagenesis processes, can also create preferential targets for environmental mutagens and carcinogens. PMID- 9354430 TI - Adenovirus-mediated overexpression of the transcription factor E2F-1 induces apoptosis in human breast and ovarian carcinoma cell lines and does not require p53. AB - Apoptosis is a mode of cell death that is carefully regulated based on cellular and environmental signals. The ability to modulate the individual cellular machinery and thereby to promote apoptosis is an important strategy in cancer therapy. It has previously been shown that overexpression of the transcription factor E2F-1 can induce apoptosis in quiescent rat embryo fibroblasts. This effect has been reported to occur in a p53-dependent manner. To investigate whether overexpression of E2F-1 could also induce apoptosis in human cancer cells, a recombinant adenovirus vector containing the transgene E2F-1 under control of the cytomegalovirus promoter (Ad5CMVE2F) was used to induce high levels of the E2F-1 protein in human breast and ovarian carcinoma cell lines. Significant morphological changes occurred in four of the five cell lines within 48 h of transduction with the Ad5CMVE2F. These changes were consistent with apoptosis, which was confirmed further by DNA fragmentation assay and fluorescence-activated cell sorting analysis. On the basis of these assays, which show that apoptosis occurred in those cell lines with mutations in the p53 gene, we suggest that the induction of E2F-1-mediated apoptosis does not require wild type p53 when E2F-1 is overexpressed using an adenovirus-based strategy. PMID- 9354432 TI - Patched (ptch)-associated preferential expression of smoothened (smoh) in human basal cell carcinoma of the skin. AB - The discovery of specific overexpression of a gatekeeper gene, ptch, in basal cell carcinoma (BCC) led to a hypothesis that the human homologue of patched (PTCH) normally functions as a negative regulator of the signaling pathway that is initiated by hedgehogs (HHs) and activated by the human homologue of smoothened (SMOH); however, no evidence for the involvement of smoh and hhs has been provided. Here, we show novel evidence that smoh is also preferentially overexpressed in BCC, together with ptch (P < 0.002), and that Sonic hh was expressed in only some BCCs. Our data, therefore, indicate that such overexpression of smoh may be associated with overexpression or mutation of PTCH and that this overexpression subsequently stimulates the PTCH/SMOH signaling pathway. In an investigation of a possible regulation of ptch and smoh, we demonstrated that expression of exogenous p21WAF1 in immortalized keratinocytes down-regulates both ptch and smoh and that the down-regulation is accompanied by growth arrest, which suggests the involvement of p21WAF1 in regulation of the PTCH/SMOH signaling pathway. PMID- 9354433 TI - PTEN/MMAC1 mutations in endometrial cancers. AB - Endometrial carcinomas represent the most common gynecological cancer in the United States, yet the molecular genetic events that underlie the development of these tumors remain obscure. Chromosome 10 is implicated in the pathogenesis of endometrial carcinoma based on loss of heterozygosity (LOH), comparative genomic hybridization, and cytogenetics. Recently, a potential tumor suppressor gene, PTEN/MMAC1, with homology to dual-specificity phosphatases and to the cytoskeletal proteins tensin and auxillin was identified on chromosome 10. This gene is mutated in several types of advanced tumors that display frequent LOH on chromosome 10, most notably glioblastomas. Additionally, germ-line mutations of PTEN/MMAC1 are responsible for several familial neoplastic disorders, including Cowden disease and Bannayan-Zonana syndrome. Because this locus is included in the region of LOH in many endometrial carcinomas, we examined 70 endometrial carcinomas for alterations in PTEN/MMAC1. Somatic mutations were detected in 24 cases (34%) including 21 cases that resulted in premature truncation of the protein, 2 tumors with missense alterations in the conserved phosphatase domain, and 1 tumor with a large insertion. These data indicate that PTEN/MMAC1 is more commonly mutated than any other known gene in endometrial cancers. PMID- 9354434 TI - Microsatellite instability analysis: a multicenter study for reliability and quality control. AB - The molecular biology section of the Hereditary Non-Polyposis Colorectal Cancer study group-Germany, instituted a multicenter study to test the reliability and quality of microsatellite instability (MSI) analysis. Eight laboratories compared MSI analyses performed on 10 matched pairs of normal and tumor DNA from patients with colorectal carcinomas. A variety of techniques were applied to the detection of microsatellite changes: (a) silver and ethidium bromide staining of polyacrylamide gels; (b) radioactive labeling; and (c) automated fluorescence detection. The identification of highly unstable tumors and tumors without MSI was achieved in high concordance. However, the interpretation of the band patterns resulted in divergent classifications at several microsatellite marker loci for a large fraction of this tumor/normal panel. The data on more than 30 primers per case suggest that the enlargement of the microsatellite panel to more than 10 loci does not influence the results. In this study, cases with MSI in less than 10% of loci were classified as microsatellite stable, whereas MSI was diagnosed in cases with more than 40% of all markers unstable. We propose that a panel of five microsatellite loci consisting of repeats with different lengths should be analyzed in an initial analysis. When less than two marker loci display shifts in the microsatellite bands from tumor DNA, the panel should be enlarged to include an additional set of five marker loci. The number of marker loci analyzed as well as the number of unstable marker loci found should always be identified. These criteria should result in reports of MSI that are more comparable between studies. PMID- 9354435 TI - Expression of three UDP-N-acetyl-alpha-D-galactosamine:polypeptide GalNAc N acetylgalactosaminyltransferases in adenocarcinoma cell lines. AB - The levels of mRNA expression of three UDP-N-acetyl-alpha-D galactosamine:polypeptide GalNAc N-acetylgalactosaminyltransferases (GalNAc transferases) were quantified for human adenocarcinoma cell lines from pancreas, colon, stomach, and breast. Two of the GalNAc-transferases, GalNAc-T1 and GalNAc T2, were expressed constitutively and at low levels in most or all cell lines examined. A third GalNAc-transferase, GalNAc-T3, was differentially expressed. Well-differentiated adenocarcinoma cell lines expressed high levels and moderately differentiated cell lines expressed lower levels of GalNAc-T3. Cell lines classified as poorly differentiated failed to express GalNAc-T3 mRNA at levels that could be detected by Northern blot analysis. Differential expression of the GalNAc-T3 protein was confirmed in these cell lines by Western blotting. We propose that glycosylation in tumor cell lines may be regulated in part by differential expression of GalNAc-transferases, and we suggest that GalNAc-T3 gene expression may be a molecular indicator of differentiated adenocarcinoma. PMID- 9354436 TI - Diagnostic microsatellite instability: definition and correlation with mismatch repair protein expression. AB - Alterations of the length of simple repetitive genomic sequences (microsatellite instability, MSI) characterize a distinct mechanism of colorectal carcinogenesis. Such MSI has been found to be associated with hereditary nonpolyposis colorectal cancer (HNPCC) that involves mutation of the human mismatch repair genes hMSH2 and hMLH1 as well as many sporadic cancers of most tissue types. Although the study of MSI status is a useful tool for HNPCC screening and for the determination of tumor prognosis in sporadic cases of colorectal cancer, the reliability of MSI diagnosis is still a subject of debate. Here we have examined 58 primary colorectal tumors (selected from a cohort of 200) using 31 microsatellite markers that comprised the most frequent simple repeat types. The expression of the hMSH2 and hMLH1 mismatch repair proteins was studied by immunohistochemistry, and most patients were surveyed for at least 2 years. Reproducibility of gel interpretation, as well as diagnostic sensitivity and specificity of the MSI status, were determined. We found that unambiguous determination of band shifts as well as MSI diagnosis were closely related to the type of the marker repeat and that MSI could be subdivided into "high" MSI (>20% unstable loci), "low" MSI (<10% unstable loci), and microsatellite stable (0% unstable loci). One-half of the patients with high MSI tumors (n = 8) fulfilled either the Amsterdam criteria (n = 4), had at least one relative with HNPCC related carcinoma (n = 2), or were diagnosed with colorectal cancer at an age below 45 years (n = 2). Fourteen of the 15 high MSI tumors had lost either hMSH2 (n = 8) or hMLH1 (n = 6) protein expression. In contrast, all of the low MSI tumors and the MSI-negative tumors displayed normal expression of hMSH2 and hMLH1. These studies provide a clear recommendation for the uniform use of a panel of 10 microsatellites and a definition of at least 40% instability (using these defined marker loci) in the diagnostic analysis of MSI. PMID- 9354437 TI - Inhibition of human cytochrome P450-catalyzed oxidations of xenobiotics and procarcinogens by synthetic organoselenium compounds. AB - The effects of synthetic chemopreventive organoselenium compounds 1,2-, 1,3-, and 1,4-phenylenebis(methylene)selenocyanate (o-, m-, and p-XSC, respectively), benzyl selenocyanate (BSC), and dibenzyl diselenide (DDS) and inorganic sodium selenite on the oxidation of xenobiotics and procarcinogens by human cytochrome P450 (P450 or CYP) enzymes were determined in vitro. Spectral studies showed that BSC and three XSC compounds (but not sodium selenite or DDS) induced type II difference spectrum when added to the suspension of liver microsomes isolated from beta-naphthoflavone-treated rats, with m-XSC being the most potent in inducing spectral interactions with P450 enzymes; m-XSC also produced a type II spectral change with human liver microsomes. o-, m-, and p-XSC inhibited 7 ethoxyresorufin O-deethylation catalyzed by human liver microsomes when added at concentrations below 1 microM levels, but BSC and DDS were less effective. All of these compounds inhibited the oxidation of model substrates for human P450s to varying extents. We studied the effects of these compounds on the activation of procarcinogens by recombinant human CYP1A1, 1A2, and 1B1 enzymes using Salmonella typhimurium NM2009 tester strain for the detection of DNA damage. The three XSCs were found to be very potent inhibitors of metabolic activation of 3-amino-1,4 dimethyl-5H-pyrido[4,3-b]indole, 2-amino-3,5-dimethylimidazo[4,5-f]quinoline, and 2-aminoanthracene, catalyzed by CYP1A1, 1A2, and 1B1, respectively. The potency of inhibition of m-XSC on CYP1B1-dependent activation of 2-aminoanthracene was compatible to those of alpha-naphthoflavone. These inhibitory actions may, in part, account for the mechanisms responsible for cancer prevention by organoselenium compounds in laboratory animals. PMID- 9354438 TI - Cyclin D1 overexpression in rat two-stage bladder carcinogenesis and its relationship with oncogenes, tumor suppressor genes, and cell proliferation. AB - Overexpression of cyclin D1 has been implicated in the malignant transformation of a variety of human cancers, including urinary bladder carcinomas. However, few reports have addressed the significance of cyclin D1 overexpression in chemical carcinogenesis in rodents. In the present study, we evaluated the oncogenic potential of cyclin D1 in experimental rat urinary bladder carcinogenesis and its relationships to the oncogenes cyclin E, K-ras, and H-ras as well as tumor suppressor genes p53 and p21WAF1/Cip1. In addition, proliferation status of preneoplastic lesions and tumors was assessed by proliferating cell nuclear antigen immunohistochemistry. Fisher 344 rats were initiated with 0.05% N-butyl-N (4-hydroxybutyl)nitrosamine in the drinking water for 4 weeks and then administered 5% sodium L-ascorbate in diet. Animals were sacrificed at weeks 4, 8, 12, 18, and 24. Preneoplastic lesions such as papillary or nodular hyperplasia and neoplastic lesions of the urinary bladder were observed during carcinogenesis. By immunohistochemical examination, overexpression of cyclin D1 protein was observed in 17% of papillary or nodular hyperplasias, 66% of papillomas, and 69% of transitional cell carcinomas, whereas nuclear accumulation of p53 was observed in none of the preneoplastic lesions and in fewer than 2% of transitional cell carcinomas. Overexpression of cyclin D1 in preneoplastic lesions and tumors was not dependent on the size of the tumors or their proliferation status. Quantitation of mRNA in tumors by multiplex reverse transcription-PCR showed that average mRNA expression of cyclin D1 and cyclin E was increased, whereas average p21WAF1/Cip1 mRNA expression was decreased. More than 2-fold overexpression of cyclin D1 mRNA was observed in 50 and 60% of tumors at weeks 18 and 24, respectively. Localization of cyclin D1 mRNA expression was demonstrated by in situ hybridization, and the results were comparable to immunohistochemistry findings. None of the 25 tumors we examined by PCR-single strand conformational polymorphism analysis harbored p53 mutations, H-ras mutations, or K-ras mutations. Thus, during the promotion phase of two-stage bladder carcinogenesis, overexpression of cyclin D1 in tumor cells may provide yet another mechanism by which tumors can gain a growth advantage. In contrast, tumors with mutated p53 may not have a growth advantage. Our results suggest that overexpression of cyclin D1 plays a critical role during urinary bladder carcinogenesis. PMID- 9354439 TI - Angiogenesis of uterine cervical carcinoma: characterization by pharmacokinetic magnetic resonance parameters and histological microvessel density with correlation to lymphatic involvement. AB - Dynamic studies of Gd-based contrast agents in magnetic resonance imaging (MRI) are increasingly being used for tumor characterization as well as therapy response monitoring. Because detailed knowledge regarding the pathophysiological properties, which in turn are responsible for differences in contrast enhancement, remains fairly undetermined, it was the aim of this project to: (a) examine the relationship between contrast-enhanced dynamic MRI-derived characteristics and histological microvessel density counts, a recognized surrogate of tumor angiogenesis, from primary or recurrent cancers of the uterine cervix; and (b) correlate these parameters with lymphatic involvement to characterize tumor aggressiveness in terms of lymphatic spread. Pharmacokinetic parameters (amplitude, A; exchange rate constant, k21) were calculated from a contrast-enhanced dynamic MRI series in 55 patients (ages 25-72 years; mean, 50 years) with biopsy-proven primary (n = 42) or recurrent (n = 13) uterine cervical cancer. Both pharmacokinetic parameters were correlated to histologically determined microvessel density counts (factor VIII-related antigen) and other pathological tumor characteristics obtained from the operative specimens after radical surgery. In addition, the magnetic resonance and histological data were correlated to the presence or absence of lymphatic system involvement. Pharmacokinetic MRI-derived parameters (A and k21) increased with increasing histological microvessel density counts with r = 0.41 and 0.50, respectively. Lymphatic involvement was more comprehensibly assessed by the pharmacokinetic parameter k21 compared with histological microvessel density, resulting in a higher sensitivity, overall accuracy, and comparable specificity. Contrast enhanced MRI parameters might prove to be applicable for estimation of tumor angiogenesis in uterine cervical cancer; thus, MRI may become an additional tool to characterize malignant progression in terms of lymphatic involvement in uterine cervical cancer. PMID- 9354440 TI - Associations of sedentary lifestyle, obesity, smoking, alcohol use, and diabetes with the risk of colorectal cancer. AB - Variation in colorectal cancer rates between countries and within ethnic groups upon migration and/or Westernization suggests a role for some aspects of Western lifestyle in the etiology of this disease. We conducted a population-based case control study in the multiethnic population of Hawaii to evaluate associations between colorectal cancer and a number of characteristics of the Western lifestyle (high caloric intake, physical inactivity, obesity, smoking, and drinking) and some of their associated diseases. We interviewed in person 698 male and 494 female United States-born or immigrant Japanese, Caucasian, Filipino, Hawaiian, and Chinese patients diagnosed in 1987-1991 with colorectal cancer and 1192 population controls matched on age, sex, and ethnicity. Conditional logistic regression was used to estimate odds ratios adjusting for dietary and nondietary risk factors. Place of birth and duration of residence in the United States were unrelated to colorectal cancer risk. Energy intake (independent of the calorie source) and body mass index were directly associated with risk, and lifetime recreational physical activity was inversely associated with risk. The associations with these factors were independent of each other, additive (on the logistic scale) and stronger in men. When individuals were cross categorized in relation to the medians of these variables, those with the higher energy intake and body mass index and lower physical activity were at the highest risk (for males, OR, 3.0; 95% confidence interval, 1.8-5.0, and for females, OR, 1.7; 95% confidence interval, 1.0-3.2). Smoking in the distant, as well as recent, past and alcohol use were directly associated with colorectal cancer in both sexes. Individuals with a history of diabetes or frequent constipation were at increased risk for this cancer, whereas past diagnosis of hypercholesterolemia was inversely associated with risk. The findings were consistent between sexes, among ethnic groups, and across stages at diagnosis, making bias an unlikely explanation. These results confirm the data from immigrant studies that suggest that the increase in colorectal cancer risk experienced by Asian immigrants to the United States occurred in the first generation because we found no difference in risk between the immigrants themselves and subsequent generations. They also agree with recent findings that suggest that high energy intake, large body mass, and physical inactivity independently increase risk of this disease and that a nutritional imbalance, similar to the one involved in diabetes, may lead to colorectal cancer. PMID- 9354441 TI - Selective tumor apoptosis by MF13, L-prolyl-L-m-[bis(chloroethyl)amino] phenylalanyl-L-norvaline ethyl ester, a new sarcolysin containing tripeptide. AB - ID50 and ID90 values for L-prolyl-L-m-[bis(chloroethyl)amino]-phenylalanyl-L norvaline ethyl ester HCl (MF13), were determined in four murine (leukemia, lymphoma, melanoma, and lung) and eight human cancer cell lines (two leukemia, prostate, kidney, colon, two melanoma, and breast). Cytotoxic activity was 2-5 times higher than that of sarcolysin [(L-3-[bis(2-chloroethyl)amino]-L phenylalanine] against all leukemias and lymphomas, ID50 0.5-0.9 microM, and against human solid tumors, ID50 0.4-2.1 microM. Sensitivities of L-phenylalanine mustard-resistant and methotrexate-resistant L1210 cells were the same as the naive lines, ID50 0.5 microM. Apoptosis was confirmed by: (a) morphology, revealing chromatin condensation and nuclear fragmentation; (b) flow cytometry, showing changes in cell size and DNA integrity; and (c) DNA electrophoresis, demonstrating multiples of 180-200-bp DNA units. MF13 had no cytotoxicity against human peripheral blood lymphocytes at concentrations lethal to tumor cells (ID50, 13.3 microM without and 11 microM with phytohemagglutinin stimulation) and failed to induce apoptosis. s.c. MF13 treatment of mice with advanced EL4 leukemic ascites yielded extensive apoptosis, with DNA degradation identical to that seen in vitro, and resulted in complete tumor regression in all treated mice. These results suggest MF13 as a potential chemotherapeutic agent. PMID- 9354442 TI - Potent antitumor activity of a novel nucleoside analogue, BCH-4556 (beta-L dioxolane-cytidine), in human renal cell carcinoma xenograft tumor models. AB - Beta-L-Dioxolane-cytidine (BCH-4556) is a novel anticancer nucleoside analogue with a stereochemically unnatural beta-L configuration. This compound was previously shown to have a potent antitumor activity in human prostate and hepatocellular xenograft tumor models (K. L. Grove et al., Cancer Res., 55: 3008 3011, 1995). Herein, we extended the efficacy validation of BCH-4556 to renal cell carcinoma (RCC) cell lines both in vitro and in vivo. In vitro cytotoxicity and proliferation inhibition determinations in human RCC cell lines CAKI-1, CAKI 2, 786-0, and A498 produced IC50 concentrations ranging from 15-35 nM. In vivo antitumor activity was consistent with the in vitro sensitivity. BCH-4556 was very effective in human RCC tumor xenograft models, including CAKI-1, A498, RXF 393, and SN12C carcinomas. Very good responses were observed in animals bearing CAKI-1, A498, and RXF-393 RCC tumors given i.p. doses of 10, 25, and 50 mg/kg twice a day for 5 days, with complete regression recorded in most of the animals tested. Curative activity was also observed, with 40-60% of animals remaining tumor free in all three RCC models at the day of study termination. Significant tumor shrinkage was also evident in the SN12C model. BCH-4556 efficacy evaluation in the orthotopic subrenal capsule tumor models demonstrated a potent tumor growth inhibition against human CAKI-1 xenografts and tumor stasis against mouse Renca tumors. BCH-4556 was also effective in inhibiting the growth of rebound CAKI-1 tumors after the administration of a second treatment cycle. The observed antitumor activity of BCH-4556 in several RCC human solid tumor xenografts, including the lethal RXF-393 model, warrants further investigation of this novel nucleoside analogue in clinical trials of RCC. PMID- 9354443 TI - Cellular pharmacokinetics and cytotoxicity of camptothecin and topotecan at normal and acidic pH. AB - pH-mediated conversions in the structure of the topoisomerase (topo) I inhibitors camptothecin (CPT) and its analogues have strong implications for the pharmacokinetics and pharmacodynamics of these novel anticancer agents. Because the cell-penetrating and biologically active lactone isomers predominate at acidic conditions, we have tested if low pH potentiates the cytotoxic and antitumor effects of CPT and its water-soluble derivative topotecan (TPT). In L1210 leukemia cells, rapid initial uptake of radiolabeled CPT and TPT was followed by a gradual release from cells at physiological pH 7.4, whereas high drug levels were maintained in cells at pH 6.2. Steady-state uptake levels of CPT increased proportionally, up to 5-fold, with decreasing pH of the incubating medium (from 7.4 to 6.0). With TPT, a maximum 3-fold increase was observed at pH 6.8 to 6.4. By contrast, the cellular pharmacokinetics of the topoisomerase II inhibitor etoposide (ETP) were independent of the ambient pH. The large increases in intracellular CPT and TPT levels caused only moderate potentiation of cytotoxicity in short-term incubations. Conditions of very low pH < or =6.2 even antagonized the cytotoxicity of the topo I and topo II inhibitors, due to inhibition of DNA synthesis by intracellular acidification. However, in clinically relevant schedules of prolonged exposures at low drug concentration, low pH potentiated the cytotoxicity of CPT and TPT by 2-3-fold. To investigate the effect of local pH in vivo, the basal interstitial pH of 6.8 of RIF-1 tumors was selectively lowered by i.p. injection of the host animals with the mitochondrial inhibitor meta-iodobenzylguanidine (32 mg/kg) and glucose (1.5 g/kg). In accordance with the pH optimum for TPT uptake at pH 6.8 to 6.4, tumor acidification had no effect on the antitumor effect of this analogue. By contrast, the intervention significantly potentiated the response of tumors to CPT. The results indicate that local pH is an important determinant of the cellular pharmacokinetics and the antitumor activity of CPT and analogues. PMID- 9354444 TI - O6-methylguanine-DNA methyltransferase (MGMT) transfectants of a 1,3-bis(2 chloroethyl)-1-nitrosourea (BCNU)-sensitive colon cancer cell line selectively repopulate heterogenous MGMT+/MGMT- xenografts after BCNU and O6-benzylguanine plus BCNU. AB - To evaluate the role of O6-alkylguanine-DNA alkyltransferase (AGT) in colon tumor chloroethylnitrosourea (CENU) resistance, AGT-deficient VACO 8 cells were transfected with a vector containing or lacking the human O6-methylguanine-DNA methyltransferase (MGMT) cDNA. VACO 8MGMT (V8MGMT) sublines possessed high levels of AGT activity in cell culture and were > 10-fold resistant to the CENU 1,3 bis(2-chloroethyl)-1-nitrosourea (BCNU). V8MGMT cells, VACO 8neo cells, and mixtures of both were grown as xenografts in nude mice. MGMT expression in VACO 8 xenografts reflected the percentage of V8MGMT cells present in the tumor inoculum. Xenografts originally containing 0-10% V8MGMT cells were sensitive to BCNU, although partial resistance was observed as the percentage of V8MGMT cells increased. Tumors containing 30-100% V8MGMT cells were completely resistant to BCNU with no regressions and no growth delays. Pretreatment with O6-benzylguanine (BG) depleted tumor AGT activity for at least 6 h and sensitized xenografts containing 1 and 100% V8MGMT cells to BCNU. After BCNU or BG + BCNU, xenografts growing from inoculums containing as low as 0.1% V8MGMT cells had high AGT activities similar to that found in V8MGMT xenografts, with the majority of the cells expressing MGMT. These results provide evidence that MGMT expression influences both intrinsic and acquired colon tumor CENU resistance, that selective expansion of AGT+ colon tumor cells commonly occurs after CENU exposure, and that BG is effective in sensitizing colon tumors to CENUs, even when only a small fraction of the cells in a heterogeneous tumor express MGMT. PMID- 9354445 TI - Systemic therapy with 3BIT, a triple combination cocktail of anti-CD19, -CD22, and -CD38-saporin immunotoxins, is curative of human B-cell lymphoma in severe combined immunodeficient mice. AB - We demonstrate in these preclinical studies that all severe combined immunodeficient mice injected with the human B-cell lymphoma cell line Ramos are cured when treated with a combination of anti-CD19, -CD22, and -CD38-saporin immunotoxins (ITs; termed 3BIT). Each component IT used individually did not cure the majority of animals but did significantly prolong their survival compared with PBS sham-treated controls, although the majority succumbed eventually to disease. The very significant improvement obtained with the three-IT combination 3BIT was not due to an antibody or antibody-plus-IT effect. We postulate that by targeting against these three cell surface molecules, we have effectively ensured delivery of saporin to each lymphoma cell with growth potential within the tumor, thus overcoming the problems of heterogeneity of target antigen expression that can limit the therapeutic efficacy of single-IT therapy or even two-IT combination therapy. These "proof of principle" findings have an obvious important bearing on antibody-based therapies for cancer and provide the rationale needed for the design and implementation of clinical trials with such combinations. PMID- 9354446 TI - Potentiation of cytochrome P450/cyclophosphamide-based cancer gene therapy by coexpression of the P450 reductase gene. AB - Intratumoral expression of cytochrome P450 2B1 sensitizes tumor cells to the cytotoxic action of the alkylating agent prodrug cyclophosphamide (CPA) and provides a novel strategy for cancer gene therapy that may enhance the selectivity and the effectiveness of this class of antitumor drugs [L. Chen and D. J. Waxman, Cancer Res., 55: 581-589, 1995]. P450-catalyzed drug metabolism is obligatorily dependent on electron input from the flavoenzyme NADPH-P450 reductase (RED), which is widely expressed in many cell types, including tumor cells. Here, we investigate the potential utility of combining RED gene transfer with CPA-based P450 gene therapy. Rat 9L gliosarcoma cells stably expressing either basal or elevated (up to 10-fold increase) levels of RED, in the presence or absence of P450 2B1, were selected and characterized. RED overexpression substantially increased the sensitivity of these cells to CPA, but only when combined with P450 2B1 expression. An enhanced cytotoxic response was also obtained when recombinant adenovirus encoding P450 2B1 was used to deliver the P450 gene to RED-overexpressing tumor cells. CPA cytotoxicity was substantially decreased by the RED inhibitor diphenyleneiodonium chloride or by the P450 inhibitor metyrapone, providing evidence of its dependence on the catalytic contributions of both protein components of the P450 metabolic pathway. Conditioned media from P450 2B1-expressing and RED-overexpressing tumor cells treated with CPA exhibited increased formation of the primary 4-hydroxy metabolite and greater cell contact-independent bystander cytotoxic potential compared to tumor cells containing P450 2B1 and basal levels of RED. Evaluation of the impact of P450/RED combination gene therapy using a s.c. solid tumor model/tumor excision assay revealed a dramatic 50-100-fold increase in tumor cell kill in vivo over that provided by liver drug activation alone. These findings establish the importance of endogenous RED levels as a determinant of the sensitivity of tumor cells to CPA/P450 gene therapy and demonstrate the striking therapeutic effectiveness of an anticancer prodrug activation strategy based on the combination of a cytochrome P450 gene with the gene encoding RED. PMID- 9354447 TI - Induction of apoptosis and cell cycle arrest by CP-358,774, an inhibitor of epidermal growth factor receptor tyrosine kinase. AB - The epidermal growth factor receptor (EGFR) is overexpressed in a significant percentage of carcinomas and contributes to the malignant phenotype. CP-358,774 is a directly acting inhibitor of human EGFR tyrosine kinase with an IC50 of 2 nM and reduces EGFR autophosphorylation in intact tumor cells with an IC50 of 20 nM. This inhibition is selective for EGFR tyrosine kinase relative to other tyrosine kinases we have examined, both in assays of isolated kinases and whole cells. At doses of 100 mg/kg, CP-358,774 completely prevents EGF-induced autophosphorylation of EGFR in human HN5 tumors growing as xenografts in athymic mice and of the hepatic EGFR of the treated mice. CP-358,774 inhibits the proliferation of DiFi human colon tumor cells at submicromolar concentrations in cell culture and blocks cell cycle progression at the G1 phase. This inhibitor produces a marked accumulation of retinoblastoma protein in its underphosphorylated form and accumulation of p27KIP1 in DiFi cells, which may contribute to the cell cycle block. Inhibition of the EGFR also triggers apoptosis in these cells as determined by formation of DNA fragments and other criteria. These results indicate that CP-358,774 has potential for the treatment of tumors that are dependent on the EGFR pathway for proliferation or survival. PMID- 9354448 TI - Monocyte chemoattractant protein inhibits the generation of tumor-reactive T cells. AB - The adoptive transfer of tumor-sensitized T cells can eradicate disseminated malignancy in murine animal models. T cells must be sensitized to tumor antigens in vivo to acquire antitumor reactivity. T-cell sensitization has been demonstrated to be dependent on host antigen-presenting cells. Tumor-associated macrophages are a heterogeneous population of cells that may have both inhibitory and stimulatory influences on the sensitization of naive T cells. Here we demonstrate that a weakly immunogenic tumor, the MCA 205 sarcoma, produces substantial amounts of murine monocyte chemoattractant protein 1 (MCP-1). Neutralization of MCP-1 during in vivo T-cell sensitization resulted in T cells that possessed enhanced therapeutic activity against established pulmonary metastases. These T cells sensitized during MCP-1 depletion also exhibited enhanced production of IFN-gamma upon recognition of tumor targets. These results demonstrate that MCP-1 can have a potent inhibitory influence on the development of tumor-reactive T cells. PMID- 9354449 TI - Human papillomavirus-specific cytotoxic T lymphocytes in patients with cervical intraepithelial neoplasia grade III. AB - Human papillomaviruses (HPVs), especially types 16 and 18, are strongly associated with the development of cervical intraepithelial neoplasia (CIN) and invasive carcinoma. The HPV E6 and E7 proteins are expressed constitutively in the majority of CIN lesions and carcinomas. Therefore, they are targets for the immune response against HPV and candidates for active immunotherapy. We have previously detected HPV-specific CTLs from the peripheral blood mononuclear cells from a cervical cancer patient following immunization with a recombinant vaccinia using in vitro restimulation with adenovirus recombinants expressing HPV16 or HPV18 E6/E7 fusion proteins. In this study, we used a similar protocol to determine the prevalence of CTL responses against HPV16 and HPV18 E6/E7 in the peripheral blood mononuclear cells from 10 CIN III and 10 normal subjects. HPV specific CTL responses were detected in 6 of 10 CIN III subjects. These CTL lines recognized HPV16 E6/E7 proteins presented by at least three MHC class 1 HLA alleles and by HPV-transformed CaSki cells. No HPV-specific CTLs were detected in normal subjects. This study demonstrates the presence of naturally occurring HPV specific memory CTLs in a majority of CIN III patients and provides an approach for further study of their role in modulating cervical malignancy. PMID- 9354450 TI - Persistent clonal proliferation of human T-lymphotropic virus type I-infected cells in vivo. AB - Clonal proliferation of human T-lymphotropic virus type I (HTLV-I)-infected cells has been detected by Southern blot analysis and inverse PCR in patients with adult T-cell leukemia, patients with HTLV-I-associated diseases, and even in asymptomatic carriers. Combining inverse PCR with long PCR, we amplified the genomic DNA regions flanking the integration sites of the HTLV-I provirus to detect clones of infected cells. Inverse long PCR revealed that increased virus load was associated with an increase of both the number of cells in each clone and the number of clones. Clonal proliferations were found in both CD4- and CD8 positive cells in a carrier and a patient with HTLV-I-associated neuropathy/tropical spastic paraparesis. These HTLV-I-infected clones persisted over several years in the same carriers, and, moreover, most of the persistent clones were CD4 positive in a HTLV-I carrier. These findings indicate that HTLV-I infection plays an important role in the clonal expansion of lymphocytes and the prolonged survival of CD4-positive cells in vivo. Surviving T-lymphocytes may be susceptible to genetic changes, leading to the onset of leukemia. PMID- 9354451 TI - CDKN2A/p16 is inactivated in most melanoma cell lines. AB - The CDKN2A gene maps to chromosome 9p21-22 and is responsible for melanoma susceptibility in some families. Its product, p16, binds specifically to CDK4 and CDK6 in vitro and in vivo, inhibiting their kinase activity. CDKN2A is homozygously deleted or mutated in a large proportion of tumor cell lines and some primary tumors, including melanomas. The aim of this study was to investigate the involvement of CDKN2A and elucidate the mechanisms of p16 inactivation in a panel of 60 cell lines derived from sporadic melanomas. Twenty six (43%) of the melanoma lines were homozygously deleted for CDKN2A, and an additional 15 (25%) lines carried missense, nonsense, or frameshift mutations. All but one of the latter group were shown by microsatellite analysis to be hemizygous for the region of 9p surrounding CDKN2A. p16 was detected by Western blotting in only five of the cell lines carrying mutations. Immunoprecipitation of p16 in these lines, followed by Western blotting to detect the coprecipitation of CDK4 and CDK6, revealed that p16 was functionally compromised in all cell lines but the one that carried a heterozygous CDKN2A mutation. In the remaining 19 lines that carried wild-type CDKN2A alleles, Western blot analysis and immunoprecipitation indicated that 11 cell lines expressed a wild-type protein. Northern blotting was performed on the remaining eight cell lines and revealed that one cell line carried an aberrantly sized RNA transcript, and two other cell lines failed to express RNA. The promoter was found to be methylated in five cell lines that expressed CDKN2A transcript but not p16. Presumably, the message seen by Northern blotting in these cell lines is the result of cross-hybridization of the total cDNA probe with the exon 1beta transcript. Microsatellite analysis revealed that the majority of these cell lines were hemi/homozygous for the region surrounding CDKN2A, indicating that the wild-type allele had been lost. In the 11 cell lines that expressed functional p16, microsatellite analysis revealed loss of heterozygosity at the markers immediately surrounding CDKN2A in five cases, and the previously characterized R24C mutation of CDK4 was identified in one of the remaining 6 lines. These data indicate that 55 of 60 (92%) melanoma cell lines demonstrated some aberration of CDKN2A or CDK4, thus suggesting that this pathway is a primary genetic target in melanoma development. PMID- 9354452 TI - Ribonucleotide reductase R2 gene expression and changes in drug sensitivity and genome stability. AB - Ribonucleotide reductase is a highly regulated, cell cycle-controlled activity that plays an important role in DNA synthesis and repair. Recent studies have shown that elevated expression of the rate-limiting R2 component of ribonucleotide reductase increases Raf-1 protein activation and mitogen-activated protein kinase activity and acts as a novel malignancy determinant in cooperation with activated oncogenes like H-ras. We show that hydroxyurea-resistant mouse L cells with elevated R2 gene expression and increased ribonucleotide reductase activity exhibit significantly decreased sensitivities to the chemotherapeutic compounds N-(phosphonacetyl)-L-aspartate (PALA) and methotrexate (MTX). Furthermore, BALB/c 3T3 cells containing a retroviral expression vector encoding the R2 sequence also showed decreased sensitivity to PALA and MTX when compared to cells containing the same vector but without the R2 coding region. Colonies that developed in the presence of PALA or MTX contained amplifications of the CAD or dihydrofolate reductase genes and exhibited wild-type p53 function as determined in sequence-specific p53 binding activity assays. NIH-3T3 cells containing the R2 retroviral expression vector also showed significantly decreased sensitivity to hydroxyurea and MTX but not to PALA. Furthermore, NIH 3T3 cells transfected with a vector containing the R2 sequence in antisense orientation exhibited increased sensitivity to hydroxyurea, PALA, and MTX. Similarly, mouse 10T1/2 cells that are highly transformed and drug resistant due to alterations in H-ras and a mutant oncogenic form of p53 exhibited significant increases in sensitivity to hydroxyurea, PALA, and MTX when transfected with a vector containing the R2 sequence in antisense orientation and compared to cells containing the same vector without the antisense sequence. These results indicate that altered expression of the R2 component is capable of significantly modifying drug sensitivity properties of tumor cells. We hypothesize that this occurs, at least in part, through a mechanism of increased genetic instability that is independent of direct p53 mutation or loss and involves R2 stimulation of the mitogen-activated protein kinase signal pathway. PMID- 9354454 TI - CD44 expression is aberrant in benign Schwann cell tumors possessing mutations in the neurofibromatosis type 2, but not type 1, gene. AB - Atypical expression of CD44 splice variants has been implicated in the progression of numerous tumors. This abnormal CD44 expression is presumed to result from gene alterations that cause tumorigenic transformation. Two tumor types that have been linked to specific gene alterations are schwannomas, which have mutations in the neurofibromatosis (NF) type 2 (NF2) gene, and neurofibromas, which characteristically possess NF type 1 (NF1) gene mutations. We examined CD44 expression in normal sciatic nerves, in schwannomas with confirmed NF2 mutations, and in neurofibromas and malignant peripheral nerve sheath tumor tissue and cell lines from NF1 patients. Compared to normal nerves, schwannomas express higher total levels of CD44 and additional splice variants, whereas CD44 expression in neurofibromas is unaltered. Malignant peripheral nerve sheath tumor tissue and cell lines express the CD44v6 epitope, which is not expressed by normal Schwann cells or by other Schwann cell tumors. These data indicate that altered CD44 expression correlates strictly with mutations in the NF2 but not NF1 gene and suggest that CD44v6 might be a marker for the malignant transformation of Schwann cells. PMID- 9354453 TI - Regulation of collagenase-3 expression in human breast carcinomas is mediated by stromal-epithelial cell interactions. AB - Collagenase-3 (MMP-13) is a recently identified member of the human matrix metalloproteinase gene family that is expressed in breast carcinomas and in articular cartilage from arthritic patients. Here, we have studied the cellular origin of this enzyme in breast carcinomas by in situ RNA hybridization, and we found that collagenase-3 is expressed by stromal cells immediately adjacent to epithelial tumor cells but not by the tumor cells themselves; nor is it expressed by the normal breast glandular epithelium. Consistent with this observation, coculture experiments using human fibroblasts and MCF-7 breast cancer cells revealed that conditioned medium from breast cancer cells stimulated the fibroblastic expression of collagenase-3 mRNA. In contrast, no stimulatory effect was observed when medium from fibroblast cells was added to breast cancer cells. These results strongly suggest that transcription of collagenase-3 in stromal cells is activated by diffusible factors released from epithelial breast cancer cells. A survey of a series of cytokines and growth factors known for their ability to induce collagenase-3 expression in human fibroblasts identified interleukin-1alpha and interleukin-1beta as potential candidates for inducing the expression of this MMP gene in breast carcinomas. According to these results, collagenase-3 should be included among the molecular factors that are detected during the stromal reaction to invasive breast cancer and that, by concerted action, may be essential for tumor growth and progression. PMID- 9354455 TI - Establishment of human peripheral lung epithelial cell lines (HPL1) retaining differentiated characteristics and responsiveness to epidermal growth factor, hepatocyte growth factor, and transforming growth factor beta1. AB - Novel human epithelial cell lines retaining characteristic features of normal peripheral airway cells were established by transfecting the SV40 large T antigen gene into primary in vitro outgrowths from normal peripheral lung specimens. These lines, designated as HPL1A to HPL1E, showed the polygonal shapes typical of epithelial cells and expressed cytokeratin in abundance. Ultrastructural examination revealed the presence of microvilli, multivesicular bodies, and multilamellar body-like structures that are characteristic of type II pneumocytes, but expression of CC1O transcripts, a highly specific marker for Clara cells, was also observed. Response to transforming growth factor beta, epidermal growth factor (EGF), and hepatocyte growth factor, all of which are thought to be important growth-regulatory molecules for cellular proliferation and developmental processes of peripheral lung, was apparent. In the HPL1A case, markedly altered cell morphology and cytoskeletal organization, potent inhibition of cell growth, and increased expression of an extracellular matrix protein were noted with transforming growth factor beta. Interestingly, both EGF and hepatocyte growth factor stimulated anchorage-dependent growth, whereas only EGF could sustain anchorage-independent proliferation. The HPL1 lines are, to our knowledge, the first series of stable epithelial lines of human peripheral lung to be described. They should be valuable for investigating various aspects of growth regulation and oncogenic processes, including the mechanisms of acquisition of anchorage independence and the interrelationships of genetic changes identified previously in lung cancers. In addition, the HPL1 lines may also prove useful for development of in vitro models for other human lung disorders as well as to elucidate the mechanisms of peripheral lung differentiation. PMID- 9354457 TI - DNA hypermethylation at the D17S5 locus in non-small cell lung cancers: its association with smoking history. AB - The aim of this study was to examine the association between DNA hypermethylation and clinicopathological features of non-small cell lung cancers (NSCLCs). The DNA methylation status at the D17S5 loci, at which a candidate tumor suppressor gene, HIC-1 (hypermethylated in cancer), was identified, of 51 paired tumor and nontumorous lung tissue specimens from NSCLC patients was examined by Southern blot analysis, using a methylation-sensitive restriction enzyme. DNA hypermethylation at this locus was found in 17 (33%) tumors and 16 (31%) nontumorous lung tissues. DNA in hypermethylation at this locus occurred more frequently in poorly than in well-differentiated tumors, especially in adenocarcinomas, and correlated significantly with the differentiation grade (P = 0.01). DNA hypermethylation at the D17S5 locus correlated significantly with the loss of heterozygosity at this locus in tumors (P = 0.01). The incidence of DNA hypermethylation was significantly higher in smokers than those who had never smoked in both tumors and nontumorous lung tissues (P = 0.03 and P = 0.01, respectively). These results suggest that DNA hypermethylation at the D17S5 locus may play a role in the development of NSCLCs in cigarette smokers. PMID- 9354456 TI - MXI1, a putative tumor suppressor gene, suppresses growth of human glioblastoma cells. AB - The Mxi1 protein functions in a regulatory network with members of the c-Myc family, in which c-Myc activates transcription and stimulates cell proliferation, and Mxi1 negatively regulates these actions. Inactivation of the MXI1 gene could, therefore, inhibit differentiation and enhance proliferation in the presence of normal levels of c-Myc, and thus MXI1 is a potential tumor suppressor gene. We and others have previously mapped the MXI1 gene to the distal portion of chromosome 10q, a region that is rearranged or affected by allelic loss in many astrocytic brain tumors. Using a newly described polymorphic CA microsatellite repeat in the third MXI1 intron, we show that 7 of 11 informative glioblastomas demonstrated MXI1 allelic loss. Sequence analysis revealed no somatic mutations in any of the six MXI1 coding exons, similar to findings in prostate tumors with MXI1 allelic loss. To determine whether MXI1 can indeed function as a suppressor of growth, we have introduced a steroid-inducible MXI1 expression vector into the U87MG cell line, a glioblastoma cell line lacking endogenous MXI1 expression. Induction of MXI1 expression resulted in a decreased growth rate and distinct morphological changes. Furthermore, cell cycle analysis demonstrated that induction of MXI1 results in accumulation of cells in the G2-M phase. Thus, these studies support the notion that MXI1 normally functions to suppress cell growth and suggest that loss of MXI1 function may play a role in human glioblastoma development. PMID- 9354458 TI - Role of intracellular redox status in apoptosis induction of human T-cell leukemia virus type I-infected lymphocytes by 13-cis-retinoic acid. AB - We have shown that cell cycle progression of human T-cell leukemia virus type I (HTLV-I)-transformed T-cell lines was inhibited by 13-cis-retinoic acid (13cRA). In the present study, we report that 13cRA inhibited proliferation and induced cell death of peripheral blood mononuclear cells obtained from four patients with acute adult T-cell leukemia but not of mitogen- or interleukin 2-activated peripheral blood mononuclear cells from HTLV-I-negative healthy donors. Because HTLV-I-infected lymphocytes are susceptible to oxidative stress, we examined the role of the intracellular redox state in 13cRA-induced cell death using a HTLV-I positive T-cell line, ATL2, as a model. 13cRA induced apoptosis in ATL2 cells within 48 h in a dose-dependent manner. The ability of 13cRA to induce apoptosis was more potent than that of all-trans-retinoic acid. Apoptosis induction by 13cRA was significantly enhanced by buthionine sulfoximine (BSO), which decreased the levels of intracellular reduced glutathione, although 13cRA by itself did not alter them, suggesting that intracellular reduced glutathione may modulate 13cRA induced apoptosis. In addition, flow cytometric analysis revealed that 13cRA increased intracellular peroxides in 24 h and that the addition of BSO further enhanced them. Although N-acetylcysteine had only a marginal effect, pretreatment with catalase markedly inhibited 13cRA-induced apoptosis. These results suggest that peroxide generation, ie., oxidative stress, may play a crucial role in the induction of apoptosis by 13cRA and further demonstrate that combined treatment with 13cRA and BSO induces apoptosis of HTLV-I-positive lymphocytes even more potently. PMID- 9354459 TI - Specific and efficient peptide substrates for assaying the proteolytic activity of prostate-specific antigen. AB - Prostate-specific antigen (PSA) is a serine protease secreted by both normal prostate glandular cells and prostate cancer cells. The major proteolytic substrates for PSA are the gel-forming proteins in semen, semenogelin (Sg) I and II. On the basis of the PSA cleavage map for Sg I and II, a series of small peptides (i.e., < or = 7 amino acids) was synthesized and coupled at the COOH terminus to 7-amino-4-methyl coumarin. Using these fluorescently tagged substrates, K(m)s and k(cat)s were determined for PSA hydrolysis, and the substrates were also tested for activity against a panel of purified proteases. Previously, a variety of chymotrypsin substrates have been used to assay the enzymatic activity of PSA. The present studies have identified a peptide sequence with a high degree of specificity for PSA (ie., no detectable hydrolysis by chymotrypsin) and improved K(m)s and k(cat)s over previously used substrates. On the basis of these parameters, the best peptide substrate for PSA has the amino acid sequence HSSKLQ. Using PC-82 human prostate cancer xenografts and human prostate tissues, this PSA substrate was used to document that prostate cancer cells secrete enzymatically active PSA into the extracellular fluid but that once in the blood, PSA is not enzymatically active. On the basis of this information, it should be possible to use the HSSKLQ peptide as a carrier to target peptide coupled prodrugs for selective activation within sites of PSA-secreting, metastatic prostate cancer cells and not within the blood or other nonprostatic normal tissues. PMID- 9354460 TI - Differential effects of synthetic nuclear retinoid receptor-selective retinoids on the growth of human non-small cell lung carcinoma cells. AB - Retinoids are promising agents for cancer chemoprevention and therapy. Nuclear retinoic acid receptors (RARs; RARalpha, -beta, and -gamma) and retinoid X receptors (RXRs; RXRalpha, -beta, and -gamma) are thought to mediate most of retinoids' effects on cell growth and differentiation. Because the majority of human non-small cell lung carcinoma (NSCLC) cell lines are resistant to all-trans retinoic acid, we searched for more potent retinoids. Therefore, we examined the effects of 37 natural and synthetic retinoids that exhibit specific binding to and transactivation of individual RARs or RXRs on the proliferation of eight human NSCLC cell lines. All of these cells expressed mRNAs of the three RXRs; however, they expressed varying levels of RARalpha and RARgamma, and only three of the eight cell lines expressed RARbeta mRNA. Cellular retinoic acid-binding proteins (CRABPs) I and II were detected in one and three of the eight cell lines, respectively. Only 8 of the 37 retinoids exhibited growth-inhibitory activity (IC50, < 10 microM) against at least two of the eight NSCLC cell lines. The active retinoids included one (TD550) of five RARalpha-selective, one (Ch55) of three RARbeta-selective, three (CD437, CD2325, and SR11364) of six RARgamma selective, and one (CD271) of four RARbeta/gamma-selective retinoids. The potency of these retinoids was low (IC50, > 1 microM), except for CD437, which was very potent (IC50, 0.1-0.5 microM). The six RXR-selective retinoids were mostly inactive even at 10 microM. However, combinations of RAR-selective and RXR selective retinoids exhibited additive effects. There appeared to be no simple correlation among the histological type of the NSCLC (adeno- or squamous), the levels of nuclear receptors or CRABPs, and the response of the cells to the growth-inhibitory effects of retinoids. Nevertheless, in contrast with former studies with natural retinoids, these results suggest that several synthetic retinoids do exhibit inhibitory activity against NSCLC cells, and some of them may be useful clinically. PMID- 9354461 TI - B lymphocytes from patients with chronic lymphoproliferative disorders are equipped with different costimulatory molecules. AB - Several costimulatory molecules play a key role in the differentiation of B lymphocytes and in T-B-cell interactions. In this study, we addressed the question of whether different receptors and counter-receptors may be expressed on malignant B lymphocytes from chronic B-cell malignancies. Using flow cytometry and reverse transcription PCR analyses, the expression of molecules belonging to the tumor necrosis factor receptor (TNFR) and tumor necrosis factor ligand (TNFL) families, as well as the expression of CD80 and CD86 molecules, was analyzed in normal B cells and in different chronic lymphoproliferative disorders of B-cell type, including B-cell chronic lymphocytic leukemia (CLL), mantle cell lymphoma, hairy cell leukemia (HCL), and HCL variant. Different patterns of expression of TNFR and TNFL superfamily molecules were demonstrated among B-cell malignancies. In particular, CD40 was commonly observed on all B cells (both tumor and normal), whereas its ligand (CD40L), which is usually undetectable on resting normal B lymphocytes, was expressed in CLL and HCL but not in other chronic lymphoproliferative disorders. CD27 was not shown in normal B cells, although it was present in all malignancies and with particularly high density in mantle cell lymphoma. CD70 was widely distributed on tumor B lymphocytes, but not on the CD5+ normal counterpart. CD30 was strongly expressed in HCL variant and weakly in B cell CLL, whereas its ligand showed a wide pattern of expression, including all neoplastic and normal B cells. TNFR II (CD120b) and CD80 were distributed on neoplastic B cells from all groups, usually at an intermediate to high degree of intensity, whereas the CD86 molecule was present at lower intensity than CD80. Finally, reverse transcription PCR analysis confirmed the presence of CD40L, CD30, and CD30L mRNAs in those B cells expressing the corresponding membrane bound proteins at low density. Our data indicate that TNFR and TNFL molecules are of use clinically both in differentiating B-cell malignancies from the normal counterpart (i.e., CD27, CD70, CD40L, CD30, and CD80) and in defining different chronic B-cell disorders (i.e., CD40L, CD27, and CD30). Interestingly, the observation that several receptors and their ligands (i.e., CD40/CD40L, CD30/CD30L, and CD27/CD70) can be expressed on the same cell suggests that these molecules play a role in initiating and maintaining the neoplastic process by mediating B-T and B-B interactions. PMID- 9354462 TI - Elimination of established murine colon carcinoma metastases by antibody interleukin 2 fusion protein therapy. AB - A recombinant humanized antibody-interleukin 2 fusion protein (huKS1/4-IL-2) was used to direct IL-2 to the tumor microenvironment and elicit a T cell-mediated eradication of established pulmonary and hepatic CT26-KSA colon carcinoma metastases in syngeneic BALB/c mice. This antitumor effect was specific because a fusion protein, which was nonreactive with these tumor cells, failed to exert any such effect. The efficacy of the huKS1/4-IL-2 fusion protein in eliminating metastases was documented because mixtures of monoclonal antibody huKS1/4 with recombinant human IL-2 were ineffective and, at best, only partially reduced tumor load. Two lines of evidence indicated the eradication of metastases and the absence of minimal residual disease in animals treated with the fusion protein: first, the lack of detection of CT26-KSA cells by reverse transcription-PCR, which can detect one tumor cell in 10(6) liver cells; and second, the tripling of life span. The effector mechanism involved in this tumor eradication is dependent on T cells because the IL-2-directed therapy is ineffective in T cell-deficient SCID mice. The essential effector cells were further characterized as CD8+ T cells by in vivo depletion studies. Such T cells, isolated from tumor-bearing mice after fusion protein therapy, elicited MHC class I-restricted cytotoxicity in vitro against colon carcinoma target cells. Taken together, these data indicate that fusion protein-directed IL-2 therapy induces a T cell-dependent host immune response capable of eradicating established colon cancer metastases in an animal tumor model. PMID- 9354463 TI - Betulinic acid triggers CD95 (APO-1/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. AB - Betulinic acid (BA), a melanoma-specific cytotoxic agent, induced apoptosis in neuroectodermal tumors, such as neuroblastoma, medulloblastoma, and Ewing's sarcoma, representing the most common solid tumors of childhood. BA triggered an apoptosis pathway different from the one previously identified for standard chemotherapeutic drugs. BA-induced apoptosis was independent of CD95 ligand/receptor interaction and accumulation of wild-type p53 protein, but it critically depended on activation of caspases (interleukin 1beta-converting enzyme/Ced-3-like proteases). FLICE/MACH (caspase-8), considered to be an upstream protease in the caspase cascade, and the downstream caspase CPP32/YAMA/Apopain (caspase-3) were activated, resulting in cleavage of the prototype substrate of caspases PARP. The broad-spectrum peptide inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, which blocked cleavage of FLICE and PARP, also completely abrogated BA-triggered apoptosis. Cleavage of caspases was preceded by disturbance of mitochondrial membrane potential and by generation of reactive oxygen species. Overexpression of Bcl-2 and Bcl-XL conferred resistance to BA at the level of mitochondrial dysfunction, protease activation, and nuclear fragmentation. This suggested that mitochondrial alterations were involved in BA-induced activation of caspases. Furthermore, Bax and Bcl-xs, two death-promoting proteins of the Bcl-2 family, were up-regulated following BA treatment. Most importantly, neuroblastoma cells resistant to CD95- and doxorubicin-mediated apoptosis were sensitive to treatment with BA, suggesting that BA may bypass some forms of drug resistance. Because BA exhibited significant antitumor activity on patients' derived neuroblastoma cells ex vivo, BA may be a promising new agent for the treatment of neuroectodermal tumors in vivo. PMID- 9354464 TI - Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemopreventive agent selenium. AB - Selenium is an essential trace element, the deficiency of which is associated with an increased incidence of some human cancers. Dietary supplementation with selenium has been reported to produce a decrease in the incidence of some cancers in humans. Thioredoxin reductase (TR) is a newly discovered homodimeric selenocysteine (SeCys)-containing protein that catalyzes the NADPH-dependent reduction of the redox protein thioredoxin (Trx). Trx is overexpressed by a number of human tumors, and experimental studies have shown that Trx contributes to the growth and to the transformed phenotype of some human cancer cells. Thus, TR, by reducing Trx, could play a role in regulating the growth of normal and cancer cells. We have investigated mechanisms by which selenium, in the form of sodium selenite, added to serum-free growth medium regulates TR activity in cancer cell lines. Selenium caused a dose-dependent increase in cellular TR activity. The increase in TR activity produced by 1 microM Se compared to medium with no added selenium was: for MCF-7 breast cancer cells, 37-fold; for HT-29 colon cancer cells, 19-fold; and for A549 lung cancer cells, 8-fold. In contrast, Jurkat and HL-60 leukemia cells showed no increase in TR activity. The half-life of the time course of induction of TR in HT-29 cells after adding selenium was 10 h. The increase in TR activity was accompanied by an increase in TR protein levels up to 3-fold and an increase in the specific activity of the enzyme of 5 32-fold, depending on the cell line. Studies using 75Se showed that the amount of selenium incorporated into TR increased with increasing selenium concentration up to a ratio of 1 selenium per TR monomer. There was an increase in TR mRNA levels of 2-5-fold at 1 microM selenium and an increase in the stability of TR mRNA with a half-life for degradation of 21 h compared to 10 h in the absence of selenium. Trx mRNA and protein levels and Trx mRNA stability were not affected by selenium. The results of the study show that the increase in TR activity caused by selenium is specific and due to several effects, including an increase in the stability of TR mRNA leading to increased TR mRNA levels, an increase in TR protein, but predominantly to an increase in the specific activity of TR associated with increased incorporation of selenium into the enzyme. PMID- 9354465 TI - The emerging role of CTLA-4 as an immune attenuator. PMID- 9354466 TI - A requirement for the Rho-family GTP exchange factor Vav in positive and negative selection of thymocytes. AB - The T cell repertoire is shaped by positive and negative selection of thymocytes that express low levels of T cell receptor (TCR) and both CD4 and CD8. TCR mediated signals that determine these selection processes are only partly understood. Vav, a GDP-GTP exchange factor for Rho-family proteins, is tyrosine phosphorylated following TCR stimulation, suggesting that it may transduce TCR signals. We now demonstrate that mice lacking Vav are viable and display a profound defect in the positive selection of both class I- and class II restricted T cells. In contrast, Vav is not essential for negative selection, though in its absence negative selection is much less effective. Vav may influence the efficiency of TCR-induced selection events by regulating the intracellular calcium flux of thymocytes. PMID- 9354467 TI - Minors held by majors: the H13 minor histocompatibility locus defined as a peptide/MHC class I complex. AB - The products of minor histocompatibility (H) loci are serious barriers to tissue transplantation even among major histocompatibility complex (MHC) identical individuals, frequently causing chronic graft rejection and graft versus host disease. Over 50 minor H loci map to mouse autosomal chromosomes but none are known at the molecular level. By expression cloning, we identified the H13 locus, a classical minor H locus first detected 30 years ago by the trait of graft rejection. The H13a allele is located on chromosome 2 and encodes a novel protein that yields the rare naturally processed nonapeptide SSVVGVWYL (SVL9) for presentation by the Db MHC class I molecule. The SVL9 peptide binds Db MHC despite the absence of the consensus binding motif, and a conservative methyl group substitution (Valine 4 <--> Isoleucine) explains why reciprocal T cell responses are elicited in H13a and H13b congenic strains. PMID- 9354468 TI - X-ray crystal structure of HLA-DR4 (DRA*0101, DRB1*0401) complexed with a peptide from human collagen II. AB - Genetic predisposition to rheumatoid arthritis (RA) is linked to the MHC class II allele HLA-DR4. The charge of the amino acid at DRbeta71 in the peptide-binding site appears to be critical in discriminating DR molecules linked to increased disease susceptibility. We have determined the 2.5 A x-ray structure of the DR4 molecule with the strongest linkage to RA (DRB1*0401) complexed with a human collagen II peptide. Details of a predicted salt bridge between lysine DRbeta71 and aspartic acid at the P4 peptide position suggest how it may participate in both antigen binding and TCR activation. A model is proposed for the DR4 recognition of collagen II (261-273), an antigen immunodominant in human transgenic mouse models of RA. PMID- 9354469 TI - Cell-autonomous defects in dendritic cell populations of Ikaros mutant mice point to a developmental relationship with the lymphoid lineage. AB - The transcription factor Ikaros is a major determinant of lymphocyte differentiation. Mice homozygous for an Ikaros dominant-negative (DN-/-) mutation lack all cells of lymphoid origin, including T, B, and natural killer (NK) cells. Mice homozygous for an Ikaros null allele lack B and NK cells but display specific defects in T lymphocytes. Nonetheless, both Ikaros mutant lines make an excess of monocytes and macrophages. Here we report that the production of subsets of antigen-presenting dendritic cells (DCs) is also defective. By constructing bone marrow chimeras, we demonstrate that the Ikaros-mediated defect in lymphocytes and DCs is intrinsic to their precursors and is not environment dependent. The selective defects in DCs manifested with the Ikaros null mutation suggest a tight linkage between development of T cells and CD8alpha+ DCs. The complete lack of DCs in the lymphoid organs of Ikaros DN-/- micke points to an essential role for the Ikaros gene family in the development of all DCs. PMID- 9354471 TI - Function of the TCR alpha enhancer in alphabeta and gammadelta T cells. AB - We have used gene targeted mutational approaches to assess the role of the T cell receptor alpha (TCR alpha) enhancer (E alpha) in the control of TCR alpha and TCR delta gene rearrangement and expression. We show that E alpha functions in cis to promote V alpha to J alpha rearrangement across the entire J alpha locus, a distance of greater than 70 kb. We also show that E alpha is required for normal alphabeta T cell development; in this lineage, E alpha is required for germline J alpha expression, for normal expression levels of rearranged V alpha J alpha genes, and for expression of a diverse V alpha repertoire. In gamma delta T cells, E alpha is not required for VdeltaDJdelta rearrangement, but, surprisingly, is required for normal expression levels of mature VdeltaDJdelta transcripts and for expression of germline J alpha transcripts. Our findings imply that E alpha function is not limited to the TCR alpha components of the TCRalpha/delta locus or to the alpha beta lineage; rather, E alpha function is important in both alphabeta and gammadelta lineage T cells. PMID- 9354472 TI - The imprint of intrathymic self-peptides on the mature T cell receptor repertoire. AB - The analysis of T cell receptor alpha (TCR alpha) chains in mice transgenic for a TCR beta chain has allowed us to demonstrate a central role for self-peptides in the positive intrathymic selection of major histocompatibility complex (MHC) class II-restricted T cells. Analysis of specific V alpha-J alpha joins in mature CD4+ TCRhigh thymocytes and in peripheral CD4+ T cells revealed a limitation in amino-acid sequences. By analysis of immature thymocytes, we could show that this limited repertoire was selected from a more diverse repertoire. By analysis of the same beta chain-transgenic mice bred to H-2Ma-deficient mice that express one or a very limited number of peptides, we could demonstrate that the V alpha-J alpha join repertoire was now altered and much more limited. Together, these data provide molecular and genetic evidence that the intrathymic positive selection of the TCR repertoire is critically affected by self-peptides presented by MHC class II molecules, most likely on thymic cortical epithelial cells. PMID- 9354470 TI - Developing lymph nodes collect CD4+CD3- LTbeta+ cells that can differentiate to APC, NK cells, and follicular cells but not T or B cells. AB - For a brief period during fetal lymph node organogenesis in mice, lymph node postcapillary high endothelial venules surprisingly express the Peyer's patch addressin MAdCAM-1. This expression allows initial seeding of this incipient structure by two unusual lymphocyte populations selectively expressing the Peyer's patch homing receptor integrin alpha4beta7: CD4+CD3- oligolineage progenitors and TCR gammadelta+ T cells. We show here that CD4+CD3- cells are lineage-restricted progenitors that express surface lymphotoxin-beta (LTbeta) and the chemokine receptor BLR1 and that can become natural killer cells, dendritic antigen-presenting cells, and follicular cells of unknown outcome, but these cells do not become T or B lymphocytes. Since the necessity of lymphotoxin in lymphoid organ development has been shown, we propose that the novel subset of CD4+CD3-LTbeta+ fetal cells is instrumental in the development of lymphoid tissue architecture. PMID- 9354473 TI - A region in the CD8 gene locus that directs expression to the mature CD8 T cell subset in transgenic mice. AB - The coreceptors CD4 and CD8 play a crucial role during thymocyte development and T cell effector function, and their expression is developmentally regulated. To determine the underlying molecular mechanisms of CD8 gene regulation we cloned the murine CD8 gene locus from genomic libraries and analyzed this region for deoxyribonuclease (DNase I) hypersensitive sites (HSS). Here we report, using transgenic mice, deletion analysis of one of the identified clusters of DNase I hypersensitivity, consisting of three DNase I-HSS and located in the intergenic region between the CD8alpha and CD8beta genes. Our data show that at least two of the DNase I-HSS constituting this cluster are individually sufficient to direct CD8alpha or heterologous transgene expression to the mature CD8 single-positive T cell subset and that this expression coincides temporally with the appearance of positively selected T cells. PMID- 9354474 TI - An enhancer that directs lineage-specific expression of CD8 in positively selected thymocytes and mature T cells. AB - Positive selection of CD4+CD8+ T cells to the CD4+CD8- helper and CD4- CD8+ cytotoxic lineages is a multistep process that involves complex regulation of coreceptor gene expression. By analyzing expression of a reporter gene in transgenic mice, we have identified a DNA segment, located between the murine CD8beta and CD8alpha genes, that has enhancer activity restricted to CD8 lineage cells. Remarkably, this enhancer functions in thymocytes undergoing positive selection to the CD4-CD8+ phenotype but not in immature double-positive thymocytes. The enhancer also functions in gut intraepithelial lymphocytes that express CD8alpha but not CD8beta, suggesting that it is specific for CD8alpha expression. The tight correlation between activation of this enhancer and the final step in positive selection has important implications for understanding the mechanism of lineage commitment in thymocytes. PMID- 9354475 TI - Distinct roles for LFA-1 and CD28 during activation of naive T cells: adhesion versus costimulation. AB - Efficient T cell activation requires the engagement of a variety of ligand/receptor molecules in addition to T cell receptor (TCR)-major histocompatibility complex (MHC)/peptide interactions. The leukocyte function antigen 1 (LFA-1) and the CD28 glycoprotein have both been implicated in T cell activation. The present study dissects the roles of LFA-1 and CD28 in the activation of naive virus-specific CD8+ T cells. We demonstrate that LFA-1 facilitates T cell activation by lowering the amounts of antigen necessary for T cell activation. In the absence of LFA-1, 100-fold more antigen was required for T cell-antigen-presenting cell (APC) conjugation and all subsequent events of T cell activation, including TCR down-regulation, Ca2+-flux, T cell proliferation, and lytic effector cell induction. Thus, LFA-1 facilitates the functional triggering of TCRs by promoting adhesion of T cells to APCs but does not affect T cell activation otherwise. In contrast, CD28 played an entirely different role during T cell activation. CD28 reduced the number of TCRs that had to be triggered for T cell activation and allowed activation of T cells by low affinity ligands. CD28 but not LFA-1 prevented induction of T cell unresponsiveness after stimulation of TCRs. These results demonstrate that LFA-1 and CD28 exhibit distinct, nonoverlapping ways to influence T cell activation and suggest that the terms costimulation and signal 2 should be revisited. PMID- 9354476 TI - The Ig alpha/Igbeta heterodimer on mu-negative proB cells is competent for transducing signals to induce early B cell differentiation. AB - The immunoglobulin alpha (Ig alpha)/Ig beta heterodimer was detected on the surface of mu-negative proB cell lines in association with calnexin. The cross linking of Ig beta on proB cells freshly isolated from bone marrow of recombination activating gene (RAG)-2-deficient mice induced a rapid and transient tyrosine-phosphorylation of Ig alpha as well as an array of intracellular proteins including Syk, PI3-kinase, Vav, and SLP-76. It also elicited the phosphorylation and activation of a MAP kinase ERK but not JNK/SAPK or p38. When RAG-2-deficient mice were treated with anti-Ig beta monoclonal antibody, developmentally arrested proB cells were induced to differentiate to the small preB cell stage as observed when the mu transgene was expressed in RAG 2-deficient mice. Thus, the cross-linking of Ig beta on proB cells appears to elicit differentiation signals analogous to those delivered by the preB cell receptor in normal B cell development. PMID- 9354477 TI - IGIF does not drive Th1 development but synergizes with IL-12 for interferon gamma production and activates IRAK and NFkappaB. AB - In these studies, IFN gamma-inducing factor (IGIF), unlike IL-12, did not drive Th1 development in BALB/c or C57BL/6 mice, but like IL-1alpha, potentiated IL-12 driven Th1 development in BALB/c mice. IGIF and IL-12 synergized for IFN gamma production from Th1 cells. Unlike IL-1alpha, IGIF had no effect on Th2 cells. IGIF signaled through IRAK, IL-1 receptor-associated kinase, to induce nuclear translocation of p65/p50 NFkappaB in Th1 cells. IL-1alpha had no effect on proliferation, cytokine production, or NFkappaB activation in Th1 cells but activated NFkappaB and proliferation in Th2 cells. Thus, Th1 and Th2 cells may differ in responsiveness and receptor expression for IL-1 family molecules. IGIF and IL-1alpha may differentially amplify Th1 and Th2 effector responses, respectively. PMID- 9354488 TI - Anaerobic work capacity in elite wheelchair athletes. AB - To study the anaerobic work capacity in wheelchair athletes, 67 elite wheelchair athletes (50 male) were studied in a 30-second sprint test on a computer controlled wheelchair ergometer during the World Championships and Games for the Disabled in Assen (1990). The experimental set-up (ergometer, protocol) proved to be adequate in terms of power output (P30, P5) velocity and heart rate. Male and female athletes were comparable with respect to personal characteristics (age, body weight, training hours). Track athletes (classified in 4 different functional classes) showed a class-related mean power output (P30: mean power produced during the 30-second sprint period) of 23, 68, 100, and 138 W for the male athletes (n = 38) and 38, 77, and 76 W for females in the upper three classes (n = 10). Sprint power was low for the group of subjects with cerebral palsy (35 W; mixed, n = 6) and relatively high for the amputee group (121 W; mixed, n = 6), female basketball players (81 W; n = 5), and two male field athletes (110 W). Significant differences between male and female athletes were found for P30 and P5 (highest mean power output over any of the six 5-second periods). As was to be expected, mean maximum heart rate in the sprint test varied significantly between the track groups from 112 (high lesion group) to 171 beats/minute(-1) (low lesion group) but not for both genders. The lower P30 in the T1 and T2 groups must be explained not only by the reduced functional muscle mass and impaired coordination but also by phenomena of cardiovascular dysfunction. Based on the performance parameters, the functional classification of the track athletes into four groups seems adequate. P30 was significantly associated with the personal characteristics of gender and hours of training. A significant correlation was found between P30 and sprint performance times for 200 meters (r = -0.79). No correlation was found between either of the forms of power output and the marathon times. Anaerobic wheelchair work capacity can be adequately studied with the 30-second sprint test that was used in this study. Anaerobic work capacity is highly variable among elite wheelchair athletes with different disabilities and from different sports disciplines and appeared quite strongly influenced by functionality, hours of training, and gender. PMID- 9354489 TI - Trunk rotatory muscle performance in post-stroke hemiplegic patients. AB - This study was undertaken to determine whether the direction of trunk rotation would have any effect on trunk muscle performance of post-stroke hemiplegic patients. The design consisted of a nonrandomized control trial in a setting of secondary care (rehabilitation unit at hospital facility). The subjects included 65 hemiplegic patients (50 males) and age-matched 80 healthy controls (38 females). Isokinetic trunk rotatory muscle performance at angular velocities of 60, 120, and 150 degrees per second was measured by using an isokinetic dynamometer (Cybex Torso Rotation Unit). There were no significant differences in the peak torque and best work between the right and left directions of the trunk rotation in the hemiplegic patients or in healthy controls, regardless of the gender of the subjects (paired t test, P > 0.05). The muscle performance of the hemiplegic patients was significantly lower than that of the controls for both genders (t test, P < 0.05). In the hemiplegic patients, the direction of trunk rotation does not affect the trunk rotatory muscle performance, although the performance itself was slightly decreased. PMID- 9354490 TI - Forces, moments, and acceleration acting on a restrained dummy during simulation of three possible accidents involving a wheelchair negotiating a curb: comparison between lap belt and four-point belt. AB - The objective of this study was to determine the effect of two types of restraining belts (lap belt and a four-point belt) on an instrumented dummy during three situations: wheelchair hitting straight into curb (SIC); wheelchair falling straight off a curb (SOC); wheelchair falling diagonally off a curb (DOC). A fully instrumented (50th percentile Hybrid III) dummy was seated in a standard wheelchair and restrained with one of the belts. The wheelchair rolled down a ramp reaching a platform at 2.4 miles per hour (comfortable walking speed). Three types of experiments were performed: SIC, SOC, DOC. Each experiment was repeated at least three times. Forces, moments, and acceleration were monitored and recorded via 48 sensors placed at the head, spine, and limbs. All experiments were videotaped and photographed. The data were averaged and compared with standards that have been previously established in car crash testing and with data recently obtained in a similar study using a nonrestrained dummy. Our results showed that in the SIC experiments, low magnitude forces, moments, and acceleration of no clinical significance were recorded with both types of belts. The wheelchair remained upright and the dummy safely seated. In the SOC experiments, the two belts prevented the dummy's ejection from the chair and, thus, have been effective in lowering the forces, moments, and acceleration and preventing significant injuries to the head and neck regions. In the DOC experiments, the lap belt proved to be somewhat more effective than the four point belt in lowering the extension forces at the upper neck and the moments at the lower neck below injury levels. It also kept the head injury criteria well below injury level. We postulate that the four-point belt was less effective because of its more extensive body fixation, which leads to concentration of moments and forces at the head and lower neck regions. The results of this study show that restraining systems can enhance the safety of wheelchair occupants in certain incidents. It has been demonstrated that the lap belt is as effective as the four-point belt system in SIC and SOC incidents. In DOC falls, neither belt could prevent falls and trauma to the head and neck region. The lap belt, however, was somewhat superior. We recommend that wheelchairs be equipped with a lap belt and patients be encouraged to buckle-up while using the wheelchair outdoors. PMID- 9354491 TI - Compliance with treatment for postpolio sequelae: effect of type A behavior, self concept, and loneliness. AB - To examine the effect of Type A behavior, self-concept, and loneliness on completion of and compliance with a postpolio sequelae treatment program, all 204 individuals who had been evaluated by the Postpolio Service were mailed the Postpolio Fatigue Questionnaire, the revised UCLA Loneliness Scale, and the Tennessee Self-Concept Scale. Patients were also asked to rate the frequency of assistive device use, their engaging in self-care activities, and requesting physical assistance from others; they had previously been administered the brief Type A Scale. Of the 46 respondents, 63% had completed the Postpolio Sequelae treatment program (completers), and 37% had either been discharged for noncompliance or refused treatment (noncompleters). Wheelchair use was significantly positively correlated with age at the time of contracting polio, number of limbs affected by polio, the Loneliness score, and months since leaving the treatment program, but significantly negatively correlated with Social Self and Family Self scores on the Tennessee Self-Concept Scale. Family Self score was significantly negatively correlated with crutch use but significantly positively correlated with asking co-workers for assistance. The frequency of taking two 15 minute breaks each day was significantly negatively correlated with a Type A score. Noncompleters reported a 61% increase in muscle weakness compared with a 1% decrease for completers. These results indicate that Type A behavior must be decreased so polio survivors complete and comply with a postpolio sequelae treatment program, be able to make necessary lifestyle changes, and possibly feel less lonely. Friends and family members must help polio survivors to accept lifestyle changes and support new assistive device use if patients are to feel valuable within their families and society and treat their postpolio sequelae. PMID- 9354492 TI - Visuospatial impairment and activities of daily living in patients with Parkinson's disease: a quantitative assessment of the cube-copying task. AB - By using a cube-copying task, visuospatial impairment in patients with Parkinson's disease was identified and studied in relation to the performance IQ, neuropsychological symptoms, and activities of daily living. In addition, a quantitative assessment of the performance IQ was also attempted by establishing a scoring guideline for the copying task. The results indicated that the subjects' performances in the copying task correlated to motor function, visuospatial impairment, and the results from the mobility and social cognition subtests of the Functional Independence Measure. The quantitative evaluation of the results from the copying task also showed a high correlation to the patients' constructional ability, assessed by the Block design and the Object assembly tasks from the Wechsler Adult Intelligence Scale-Revised. This study demonstrated that visuospatial deficits in Parkinson's disease patients were associated with impairments in both activities of daily living and motor function. In addition, quantitative assessment of the copying task suggested that this test may be predictive of the nonverbal IQ. Therefore, we concluded that the copying task is a useful tool for the assessment of visuospatial impairment in patients with Parkinson's disease; these deficits may be associated with an increased risk of motor dysfunction such as difficulties in the activities of daily living. PMID- 9354493 TI - Spontaneous electromyographic potentials in cervical cord-injured patients are related to dysesthetic pain. AB - A total of 61 traumatic cervical cord-injured patients were included in this study. Needle electromyography and nerve conduction study were performed at 6 to 24 weeks postinjury. Correlation between the presence of spontaneous electromyographic potentials and the presence of dysesthetic pain, as well as other clinical characteristics including age, gender, level of injury, severity of injury, spasticity, duration of injury, and performance of spinal surgery was statistically analyzed. Of the 31 patients who had spontaneous electromyographic potentials in their hands, 27 (87%) had dysesthetic pain in their limbs. Only 9 (30%) of the other 30 patients without spontaneous potentials developed dysesthetic pain. A significant correlation (P < 0.001) between the presence of spontaneous electromyographic potential and dysesthetic pain was found. The presence of spontaneous electromyographic potentials was also significantly (P < 0.05) correlated with severity of injury but not with age, gender, injury level, duration of injury, operation, and spasticity. In conclusion, the presence of spontaneous electromyographic potentials in cervical cord-injured patients was significantly related to the presence of dysesthetic pain. They occurred more often in patients with more severe injury. PMID- 9354494 TI - Changes in impairment and disability from the third to the sixth month after stroke and its relationship evaluated by an artificial neural network. AB - Functional recovery in a rather late stage after stroke was examined in 70 stroke patients using the Stroke Impairment Assessment Set (SIAS) and the Functional Independence Measure (FIM). The SIAS and the FIM were administered at three and six months after the onset of stroke. Motor items and the abdominal manual muscle testing item improved in more than 30 percent of patients. The motor subscore of the FIM changed from 60.8 to 73.4, and the cognitive subscore changed from 28.4 to 30.4. The relationship between impairment and disability was evaluated using the neural network method with the software, Skiltran. The change of the FIM motor subscore from three months to six months was used as an output variable, and the change in the SIAS items and the FIM motor subscore were included as input variables. As a result of the connection weight obtained from this network, the change in the fifth motor item (one of the tone items, abdominal manual muscle testing) and the unaffected side grip as well as the FIM at three months had a strong connection to the change of the FIM. It is compatible with ordinary experience that function of both the affected and unaffected side influences the level of disability. Contribution of the impairment to the disability indicates the importance of taking into consideration the impairment for predicting prognosis and selecting adequate treatment when we carry out stroke rehabilitation. In conclusion, we described the relationship between the SIAS and the FIM using the neural network in stroke patients and proved the importance of the impairment to predict the outcome of disability. PMID- 9354495 TI - Comparison of balance responses and motor patterns during sit-to-stand task with functional mobility in stroke patients. AB - This study was undertaken to explore whether we could provide supportive laboratory evidence for the clinical observations that a stroke patient has lost functional mobility/locomotion capability based on dynamic balance responses (center of force sway patterns) and motor control activities (electromyography patterns) during the motor task of sit-to-stand. A computerized controlled dynamic postural control assessment system was developed and used in this study. Various dynamic balance indexes were introduced and derived from center of force sway patterns expressed in four domains (space, time, force, and frequency). Motor control was assessed by multichannel surface electromyography of each side of the lower limb during the same motor task. The functional mobility capability was evaluated using the traditional FIM method. Fourteen stroke patients with right hemiplegia and nine healthy elderly individuals were recruited as the experimental and control groups, respectively. Muscle activity was recorded for quadriceps, hamstrings, anterior tibialis, and triceps surae muscles and was used for analysis. Center of force sway patterns and ground reaction forces were registered. All signals were synchronized at "seat-off." Surface electromyographic patterns of activities recorded during sit-to-stand and dynamic balance indexes computed from center of force sway patterns were categorized and compared with the functional mobility scores. Results show that both the motor control patterns and dynamic balance indexes correlated well to the extent of mobility impairment evaluated using the traditional FIM method. An important conclusion for rehabilitation medicine is that the functional mobility capability of stroke patients may be expressed numerically using dynamic balance indexes and visualized graphically through electromyographic motor patterns. PMID- 9354496 TI - Rehabilitation outcomes of patients who have developed Guillain-Barre syndrome. AB - The objective of this study was to determine associations between early variables (requirement for ventilator support, anemia, indicators of abnormal peripheral nerve function (proprioception, vibratory, fine touch/pinprick, deep-tendon reflexes, cranial nerve involvement, dysautonomia, electrodiagnostic findings), plasmapheresis, age, and gender) and outcome variables (length of acute hospitalization, length of inpatient rehabilitation, Functional Independence Measure (FIM) Rasch converted scores, and acute and rehabilitation charges) in Guillain Barre Syndrome (GBS). The design of the study was a retrospective case review of 39 GBS admissions (as defined by National Institute of Neurologic Disorders and Stroke clinical criteria) to an inpatient rehabilitation unit at a university tertiary care rehabilitation center during a three-year period. The average length of stay for 39 patients requiring transfer to the inpatient rehabilitation unit (40% of all acute care GBS admissions) was 34 days in acute care and 26 days in rehabilitation. The average adjusted charges for inpatient rehabilitation (1993 dollars) was $31,636.28. Those who required ventilator support before rehabilitation v those who did not had an admission mean FIM Rasch converted motor score of 26.6 v a score of 38.3 (P = 0.0469), gained only 10.3 points on their FIM Rasch converted motor score v 27.7 points (P = 0.0001), and had a mean acute length of stay of 66.2 days v 19.3 days (P = 0.0029). Patients requiring ventilator support were more likely to have dysautonomia (P = 0.0009). Thirty-one of 39 patients with GBS (79%) had anemia. No correlation was found between hematocrit or hemoglobin and motor function recovery as assessed via the Rasch transformed FIM motor scores. There was an association between autonomic dysfunction and an increased acute care length of stay (P = 0.0325) and total length of hospital stay (P = 0.0203). Cranial nerve dysfunction resulted in an increase in the acute care length of stay (P = 0.0266), the total length of hospital stay (P = 0.0123), and adjusted hospital charges while undergoing inpatient rehabilitation (P = 0.0235). For patients with GBS necessitating admission to inpatient rehabilitation, the requirement of prior ventilator support most strongly predicts an extended length of stay for inpatient rehabilitation. PMID- 9354497 TI - Three-dimensional pushrim forces during two speeds of wheelchair propulsion. AB - Upper limb pain frequently occurs in manual wheelchair users. Analyzing the pushrim forces and hub moments occurring during wheelchair propulsion is a first step in gaining insight into the cause of this pain. The objectives of this study were as follows: to describe the forces and moments occurring during wheelchair propulsion; to obtain variables that characterize pushrim forces and are statistically stable; and to determine how these variables change with speed. Convenience samples (n = 6) of paralympic athletes who use manual wheelchairs for mobility and have unimpaired arm function were tested. Each subject propelled a standard wheelchair on a dynamometer at 1.3 and 2.2 m/s. Biomechanical data were obtained using a force- and moment-sensing pushrim and a motion analysis system. A number of variables that describe the force and moment curves were evaluated for stability using Cronbach's alpha. Those measures found to be stable (alpha > 0.8) at each speed were then examined for differences associated with speed. The tangential, radial, and medial-lateral forces were found to comprise approximately 55, 35, and 10% of the resultant force, respectively. In addition to duration of stroke and propulsion, the following variables were found to be stable and to differ with speed (1.3 m/s +/- SD; 2.2 m/s +/- SD): peak force tangential to the pushrim (45.9 +/- 17.9 N; 62.1 +/- 30 N), peak moment radial to the hub (9.8 +/- 4.5 N x m 13.3 +/- 6 N x m), maximum rate of rise of the tangential force (911.7 +/- 631.7 N/sec; 1262.3 +/- 570.7 N/sec), and maximum rate of rise of the moment about the hub (161.9 +/- 78.3 N x m/s; 255.2 +/- 115.4 N x m/s). This study found stable parameters that characterize pushrim forces during wheelchair propulsion and varied with speed. Almost 50% of the forces exerted at the pushrim are not directed toward forward motion and, therefore, either apply friction to the pushrim or are wasted. Ultimately, this type of investigation may provide insight into the cause and prevention of upper limb injuries in manual wheelchair users. PMID- 9354498 TI - Are transitional year programs obsolete because of proposed changes in graduate medical education funding? A commentary. PMID- 9354499 TI - Compliance by physical medicine and rehabilitation residency applications with the Americans with Disabilities Act, the Civil Rights Act of 1964, and the Rehabilitation Act of 1973. A commentary. PMID- 9354501 TI - Disability Evaluation Certificate Program. PMID- 9354500 TI - Are standardized outcome measures valid for small rehabilitation units? A commentary. PMID- 9354502 TI - Analysis of pulmonary surgery data: the next step. PMID- 9354503 TI - Minimally invasive coronary artery bypass surgery: really minimal? PMID- 9354504 TI - Embolotherapy of large pulmonary arteriovenous malformations: long-term results. AB - BACKGROUND: The purpose of this study was to document the long-term results of transcatheter embolotherapy of large pulmonary arteriovenous malformations (PAVMs). METHODS: From a data base of 221 consecutive patients with PAVMs treated by embolotherapy between 1978 and 1995, 45 patients with 52 PAVMs, supplied by feeding arteries 8 mm in diameter or larger, were selected for a retrospective investigation. RESULTS: Of 45 patients with 52 large PAVMs, 38 patients (84%) with 44 PAVMs (85%) were cured by the first embolotherapy (mean follow-up, 4.7 years). Acute periprocedural complications included self-limited pleurisy (31%), angina secondary to air embolus (2%), and paradoxical embolization of a device during deployment (4%). None of these events led to short- or long-term sequelae. Seven patients (16%) had persistence of the PAVM attributable to either recanalization (n = 4) or interim accessory artery growth (n = 3). Two of these patients presented with ischemic stroke several years after the initial treatment. Persistent PAVMs (n = 8) were retreated successfully by a second procedure (n = 7), or a third procedure (n = 1) (mean follow-up, 5.9 and 5.3 years, respectively). CONCLUSIONS: Embolotherapy of large PAVMs results in permanent occlusion in an overwhelming majority of patients. Continued patency due to recanalization or accessory artery growth is easily detected and treated. PMID- 9354505 TI - Pleural cytologies in lung cancer without pleural effusions. AB - BACKGROUND: Malignant pleural effusions significantly increase the stage of lung cancer with attendant worsening of prognosis. There is a paucity of literature evaluating malignant pleural lavage cytology in patients without pleural effusions. We propose to determine the incidence of malignant pleural cytologies in patients without pleural effusions who undergo curative resection for lung cancer and to identify any predictive risk factors for positive cytology. METHODS: Seventy-eight patients underwent curative resection for lung cancer. Lavage was performed before lung manipulation and after resection and cytologically evaluated. RESULTS: Twelve pneumonectomies, 64 lobectomies, and 2 wedge resections were performed on 40 men and 38 women with an average age of 65.7 years. Fourteen percent had positive lavage cytology before lung resection with an 11% (6 of 53) incidence in stage I. A significant correlation to adenocarcinoma compared with squamous cell was found (p = 0.03) but not to stage, T or N status, grade, pleural invasion, or preoperative transthoracic needle biopsy. CONCLUSIONS: The incidence of positive pleural cytology in otherwise stage I patients is disconcertingly high. Positive cytology may be a prognosticator of a more aggressive tumor biology. PMID- 9354506 TI - Mycophenolate mofetil for obliterative bronchiolitis syndrome after lung transplantation. AB - BACKGROUND: The development of obliterative bronchiolitis after lung transplantation portends a poor long-term outcome because of progressive decline in allograft function. There are currently no effective means of treating this condition. METHODS: Thirteen patients in whom obliterative bronchiolitis syndrome developed after lung transplantation were treated with mycophenolate mofetil, an antimetabolite immunosuppressant, at a dose of 1.5 g orally twice daily. Patients were followed up clinically and with pulmonary function testing. RESULTS: Duration of mycophenolate mofetil therapy ranged from 1 week to 24 months (mean duration, 11.4 months). Pulmonary function test results stabilized in the majority of patients with no significant further decline in forced expiratory volume in 1 second. Two patients died of progressive obliterative bronchiolitis, 1 patient is alive with progressive disease, and 1 patient died of an acute infection. The drug was discontinued in 2 additional patients. In no patient did severe leukopenia or cytomegalovirus infection develop; 1 patient had a fungal infection, and 7 patients experienced gastrointestinal side effects. CONCLUSIONS: In the setting of obliterative bronchiolitis syndrome, mycophenolate mofetil is generally well tolerated and is associated with stabilization of pulmonary function test results. These findings suggest that the otherwise progressive process of obliterative bronchiolitis can be slowed. PMID- 9354507 TI - Loss of alpha-v integrin expression and recurrence in node-negative lung carcinoma. AB - BACKGROUND: Despite "curative" resection, metastases develop in many patients with node-negative (N0) non-small cell lung carcinoma. Alternative biologic markers for this tumor would be useful. Integrins are cell adhesion molecules that are thought to be important in tumor progression, and expression of these molecules previously has been shown to be altered in non-small cell lung carcinoma. We evaluated alterations in integrin expression and clinical outcome. METHODS: Immunohistochemical staining of tumor specimens was performed, and clinical data were reviewed retrospectively. RESULTS: Data were complete for 42 patients. Half of all patients (21/42) and 9 of 26 patients with negative nodes experienced tumor recurrence during follow-up. Neither histologic type nor tumor differentiation status correlated with recurrence. However, loss of the alpha v integrin subunit was associated significantly with recurrence in the N0 group. Seventy-five percent of patients with negative nodes who exhibited recurrence lost alpha v expression, compared with only 10% of patients with negative nodes who did not exhibit recurrence (p = 0.012). Alterations of other integrin subunits did not correlate significantly with prognostic follow-up variables. CONCLUSIONS: Loss of alpha v expression may serve as a marker for patients with node-negative non-small cell lung carcinoma who are at high risk for recurrence, potentially directing additional therapies. PMID- 9354509 TI - Cardiopulmonary function at rest and during exercise after resection for bronchial carcinoma. AB - BACKGROUND: Measurements of postoperative spirometric values after pneumonectomy and lobectomy vary considerably, and few researchers have studied the changes in exercise capacity during maximal work after lung resection. The purpose of this study was to describe the postoperative alterations in cardiopulmonary function. METHODS: Ninety-seven consecutive patients with lung malignancy were prospectively examined with maximal exercise test, spirometry, and arterial gas tensions. Fifty-seven patients were reinvestigated 6 months postoperatively. RESULTS: In patients having lobectomy, forced expiratory volume in 1 second decreased 8%, and exercise capacity, expressed by maximal oxygen uptake and maximal work rate, significantly decreased 13%. In patients having pneumonectomy forced expiratory volume in 1 second significantly decreased 23%, but the loss in lung volume was partly compensated as measured by exercise capacity, which decreased only 16%. Generally patients with the smallest preoperative forced vital capacity had the smallest postoperative deterioration expressed in percentages. We found a weak correlation between alterations in maximal oxygen uptake and lung function after resection. CONCLUSIONS: Lobectomy is associated with only minor deterioration of lung function and exercise capacity. Pneumonectomy causes a decrease in pulmonary volumes to about 75% of the preoperative values, partly compensated in better oxygen uptake, which postoperatively was about 85% of the preoperative values. Alteration in forced expiratory volume in 1 second is a poor predictor of change in exercise capacity after pulmonary resection. PMID- 9354508 TI - Transsternal closure of bronchopleural fistula after pneumonectomy. AB - BACKGROUND: Bronchopeural fistula after pneumonectomy, with associated empyema, has no standard therapy. The transsternal, transpericardial approach was used in all patients presenting with a large fistula. METHODS: From 1974 through 1995, 55 patients underwent transsternal, transpericardial closure of a bronchopleural fistula. Mean age was 62.7 years (range, 33 to 78 years). Malignant disease had been the indication for pneumonectomy in 50 patients and benign lesions in 5 patients. The fistula was right-sided in 41 patients (74.5%), and the bronchial stump was less than 2 cm in 25 (45.5%). Treatment of the concomitant empyema was by closed drainage in 2 patients, by repeated needle aspiration in 17, and by open thoracostomy in 36 patients. Reamputation and closure of the stump was possible in 51 patients; in 4 a primary carinal resection was done. RESULTS: Three patients died within 30 days after operation (5.4%, 70% confidence interval 2.4%-10.7%). Ten patients died late during hospitalization, total hospital mortality, 23.6% (70% confidence interval 17.3% to 31.0%). Recurrent fistula symptoms were caused by a large recurrency in 6 patients (all died), by a small one in 7 (one death due to pulmonary embolism). Mean duration of hospital stay was 56 days (range, 2 to 174 days). At follow-up of 42 patients, there were no recurrent fistulas. All patients with benign lesions are alive and well. Of 37 cancer patients, 29 died, more than half due to malignancy. Risk factors for death included recurrent fistula, short interval between pneumonectomy and onset of fistula, and closing technique. Risk factors for recurrent fistula were a short bronchial stump and the nonuse of an open thoracostomy. CONCLUSIONS: Long term results of transsternal closure are good, but hospital mortality is high. The present treatment of patients with large postpneumonectomy bronchopleural fistula includes early open thoracostomy, improvement of nutritional status, transsternal closure using resorbable sutures, and closure of the pleural space 3 weeks later. PMID- 9354510 TI - Porous-type tracheal prosthesis sealed with collagen sponge. AB - BACKGROUND: Reconstruction of a long section of the trachea is clinically problematic. Tracheal reconstructions using prostheses have met with limited success due to local infection, hemorrhage, luminal stenosis and prosthesis dislocation. METHODS: We have designed a porous type of tracheal prosthesis in which the mesh is sealed with collagen sponge. We used this prosthesis (50 mm in length) to reconstruct the cervical trachea in 10 mongrel dogs and evaluated its efficacy. RESULTS: One dog died due to an accident with anesthesia at 6 weeks and 1 of suffocation at 10 weeks. The other 8 dogs had an uneventful postoperative course until they were killed between 6 and 24 months after implantation. At sacrifice, all the prostheses had become completely incorporated into the host. Microscopic examination revealed advanced formation of a new epithelial lining in 1 dog at 6 months, and a confluent epithelial lining was observed in another dog at 12 months. Central stenosis was not significant in any of the animals. CONCLUSIONS: This tracheal prosthesis gives good results in canine tracheal reconstruction, and appears very promising for the clinical repair of tracheal defects. PMID- 9354511 TI - Gangrene of the lung: treatment in two stages. AB - BACKGROUND: Pulmonary gangrene is a rare complication of severe lung infection with devitalization of lung parenchyma and secondary infection. If untreated, gangrene of the lung leads to sepsis, multiple-organ failure, and death. Resection of all gangrenous tissue is mandatory and is lifesaving. Pleural empyema commonly accompanies gangrene of the lung; in its presence, dissection of hilar structures for resection can lead to mediastinitis or bronchopleural fistula and should be avoided. METHODS: Three patients with pulmonary gangrene were treated in two stages: immediate fenestration first and then delayed resection of gangrenous lung in a clean field and immediate closure of the pleural window. RESULTS: Two patients underwent pneumonectomy and 1 patient, lobectomy. All patients recovered without complications. CONCLUSIONS: Creation of a pleural window (fenestration) for 1 week enables safe and curative resection of a gangrenous lung or lobe in a clean field and in a patient in stable condition. PMID- 9354512 TI - Thoracoscopic sympathectomy for upper limb hyperhidrosis: looking for the right operation. AB - BACKGROUND: Thoracoscopic sympathectomy is the most effective treatment for upper limb hyperhidrosis. However, this is offset by the occurrence of a high rate of side effects, such as embarrassing compensatory sweating. Anticipating that a technique that respects the sympathetic chain and divides only the rami communicantes may lead to fewer side effects, we assessed the technique described by R. Wittmoser, comparing it with conventional thoracoscopic sympathecomy. METHODS: A total of 240 thoracoscopic sympathectomies were performed in 124 patients suffering from upper limb hyperhidrosis. Fifty-four patients underwent a conventional sympathectomy (group TS), 62 underwent division of the rami communicantes with respect to the main trunk (group SS), and 8 underwent both procedures (group TS/SS) because of accidental division of the chain during dissection. The mean follow-up is 8 months. RESULTS: No recurrence was observed in group TS whereas six (5%) occurred in group SS (p < 0.05). The global rate of compensatory sweating was about the same in both groups: 72.2% in group TS and 70.9% in group SS. However, the rate of embarrassing or disabling compensatory sweating was significantly higher in group TS (50%) than in group SS (21%) (p < 0.001). CONCLUSIONS: Although selective division of the rami communicantes results in a significant decrease in the rate of disturbing side effects, it also leads to recurrences that are usually not observed at that level in patients treated with the conventional technique. Therefore other means of achieving the ideal operation should be explored, that is, a technique associated with a high success rate but a minimal number of side effects. PMID- 9354514 TI - The role of tracheostomy in acquired immunodeficiency syndrome. AB - BACKGROUND: Tracheostomy tube (TT) insertion for respiratory failure in patients with acquired immunodeficiency syndrome has been associated with an early mortality rate of 100%. We have reviewed our experience with tracheostomy to determine if there is a role for this procedure among certain subgroups. METHODS: A retrospective review was conducted of 47 patients diagnosed with acquired immunodeficiency syndrome who underwent tracheostomy from 1988 to 1995. Patients were divided into three groups based on indications for tracheostomy: group 1, Pneumocystis carinii pneumonia (PCP); group 2, non-PCP pneumonia; and group 3, others (including neurosyphilis, endocarditis, and trauma). RESULTS: All groups were similar with regard to demographic details and laboratory values (mean age, 38 +/- 1.4 years; 95% male; CD4 count = 21.8 +/- 3.6 cells/microL). In the vast majority of cases the decision to place a TT was elective. Forty-three percent of all patients had signed do not resuscitate orders before endotracheal tube intubation. The mean time from endotracheal tube to TT insertion was 14.1 +/- 1.6 days. Early mortality after TT placement was dismal (91%) for group 1 patients but significantly better (47%) in group 2 patients (p = 0.04). Early mortality usually occurred within 3 weeks of TT placement (range, 1 to 54 days). The cause of pneumonia (PCP versus non-PCP) was the only statistically significant variable in predicting outcome. For those who survived to TT removal (26%), the average time to removal of TT was 67 +/- 11 days. Long-term survival was noted in 8 group 2 patients (mean, 584 days) and in 2 group 1 patients (450 days). CONCLUSIONS: Outcome after tracheostomy in patients with AIDS is generally poor. Patients with PCP should not undergo TT placement; however, patients with non-PCP pneumonia have a reasonable expected survival and should undergo the operation. PMID- 9354513 TI - Effect of fibrin glue in the reduction of postthoracotomy alveolar air leak. AB - BACKGROUND: Intraoperative use of fibrin glue has been advocated in reducing postthoracotomy alveolar air leak, although most studies have not been randomized and have focused on its routine use after lung resection. METHOD: This study specifically addresses the effectiveness of fibrin glue in reducing alveolar air leak only in patients considered intraoperatively to have continued moderate to severe alveolar air leak after all conventional measures to reduce it have been used. RESULTS: During a 24-month period, 66 patients undergoing lobectomies, segmentectomies, or decortication were randomized either to serve as controls (n = 33) or to have fibrin glue sprayed on the "raw" lung surface (n = 33). The median duration of intercostal drainage and in-hospital stay was 6 and 9 days, respectively, in the control group and 6 and 8 days, respectively, when fibrin glue was used. Statistical analysis revealed no differences between the groups. CONCLUSION: Fibrin glue does not add to conventional techniques in reducing moderate to severe alveolar air leak after thoracic operations. PMID- 9354515 TI - Reoperations on the aortic root and ascending aorta. AB - BACKGROUND: Reoperations on the aortic root and the ascending aorta are being performed with increasing frequency and remain a challenging problem. METHODS: Eighty-one patients (age range, 14 to 78 years) underwent reoperations on the aortic root or the ascending aorta during a 14.5-year interval ending October 1996. The previous operations were aortic valve procedure (28%), coronary artery bypass grafting (25%), aortic root replacement (24%), ascending aortic replacement (19%), and miscellaneous (5%). Twenty-two patients (27%) had had two or more previous operations. The principal indications for reoperation were true or false aneurysm (35%), acute or chronic dissection (28%), and malfunction of an aortic valve substitute (27%). The reoperations performed were aortic root replacement (composite graft, allograft, or autograft) in 48 patients and graft replacement of the ascending aorta in 33 patients. Concomitant procedures included aortic arch replacement in 43 patients (55%) and coronary artery bypass grafting in 33 patients (41%). RESULTS: The 30-day mortality rate was 8.6% (7 patients). It was 8.3% for aortic root replacement and 9.1% for ascending aorta replacement (p > 0.05). Using stepwise multivariate logistic regression analysis of 23 variables, preoperative functional class III/IV (p = 0.047) and duration of cardiopulmonary bypass (p = 0.007) were significant independent predictors of early death. The mean duration of follow-up was 3.6 years. The 1-year, 5-year, and 10-year survival rates were 89%, 81%, and 69%, respectively. Freedom from reoperation on the heart or ascending aorta was 98%, 92%, and 69%, respectively. Reoperation for false aneurysm (p = 0.050) and the presence of coexisting coronary artery disease requiring bypass grafting (p = 0.010) were the only significant predictors of late mortality. CONCLUSIONS: Reoperations on the aortic root and the ascending aorta can be accomplished with acceptable early mortality and satisfactory long-term results. More frequent resection of the aneurysmal or dissected segments of the ascending aorta and aortic root at the initial operation may reduce the need for subsequent reoperation. PMID- 9354516 TI - Vascular endothelial growth factor attenuates myocardial ischemia-reperfusion injury. AB - BACKGROUND: Hypoxic endothelial cell activation plays a key role in the myocardial dysfunction resulting from ischemia-reperfusion injury. Recent evidence suggests that vascular endothelial growth factor (VEGF) may, in addition to promoting angiogenesis, modulate various aspects of endothelial function and repair. We examined whether administration of VEGF in the cardioplegic solution might have a beneficial effect on myocardial ischemia-reperfusion injury in an isolated rat heart model. METHODS: Hearts from Sprague-Dawley rats were perfused with Krebs-Henseleit solution in a modified Langendorff apparatus. Percent recovery of cardiac output, coronary flow, stroke work, and percent increase in coronary vascular resistance were measured after 2 hours of global ischemia and 40 minutes of reperfusion. Coronary effluent was collected after ischemia and reperfusion for measurement of creatine kinase. RESULTS: Hearts receiving cardioplegia solution containing 125 microg VEGF showed significantly improved recovery of cardiac output, coronary flow, and stroke work, and significantly reduced coronary vascular resistance compared with hearts receiving hyperkalemic cardioplegia only (p < 0.05). Coadministration of a nitric oxide synthase inhibitor attenuated the VEGF-induced cardiprotective effects. Hearts treated with VEGF released significantly less creatine kinase compared with control hearts. CONCLUSIONS: Addition of VEGF to hyperkalemic cardioplegia protects against myocardial ischemia-reperfusion injury in the isolated rat heart. PMID- 9354517 TI - Clinical markers in CSF for determining neurologic deficits after thoracoabdominal aortic aneurysm repairs. AB - BACKGROUND: Spinal cord ischemia is a major cause of morbidity and mortality after thoracoabdominal aortic aneurysm operations. The incidence of paraplegia is high even in experienced institutions. METHODS: We investigated whether neurotransmitter excitotoxicity is associated with neurologic deficits after thoracoabdominal aortic aneurysm operations. We hypothesized that patients with spinal cord injury would manifest elevated levels of excitatory amino acids in their cerebrospinal fluid. Sixteen patients undergoing thoracoabdominal aortic aneurysm operations had cerebrospinal fluid drawn through lumbar spinal drains preoperatively, intraoperatively, and postoperatively. Excitatory amino acid levels (glutamate, aspartate, glycine) were measured using high-performance liquid chromatography. Excitatory amino acid levels were compared in patients who exhibited no neurologic deficits postoperatively (group I; n = 12) with patients who had clinically evident lower extremity and cerebral neurologic deficits (group II; n = 4). RESULTS: Significant elevations in glutamate and aspartate levels from baseline (p < 0.05) were limited to group II. Excitatory amino acid levels in group II were significantly elevated (p < 0.05) compared with those observed in group I. Glutamate levels were especially increased during aortic cross-clamping and late reperfusion, whereas aspartate levels were increased only during late reperfusion. CONCLUSIONS: These data suggest that neurotransmitter excitotoxicity plays a significant role in central nervous system injury. PMID- 9354518 TI - Effects of increased ICAM-1 on reperfusion injury and chronic graft vascular disease. AB - BACKGROUND: The purpose of this study was to assess the impact of increased donor cardiac intercellular adhesion molecule (ICAM-1) expression on both reperfusion injury and chronic graft vascular disease after transplantation. METHODS: Hearts were harvested from donor rats before and after pretreatment with lipopolysaccharide at -24 hours, underwent 45 minutes of cold ischemia, and were transplanted into ACI recipients with or without anti-ICAM-1 monoclonal antibody treatment. Grafts were procured early for analysis of ICAM-1 expression and reperfusion injury or the recipients were treated with cyclosporin A (to allow long-term graft acceptance) for postoperative days 0 through 9 with procurement on postoperative day 90 to histologically score for chronic graft vascular disease. RESULTS: Lipopolysaccharide-pretreated PVG heart grafts showed increased ICAM-1 expression by Northern blot and immunohistochemical analysis leading to increased reperfusion injury as assessed by neutrophil infiltration (myeloperoxidase), cardiac edema (percentage wet weight), and histologic injury (percentage area of contraction band necrosis), which was reversed by recipient treatment with anti-ICAM-1 monoclonal antibody. After administration of cyclosporin A, 5 mg/kg for 10 days, lipopolysaccharide-treated grafts had significantly worse chronic graft vascular disease scores (2.56 +/- 0.57 versus 1.84 +/- 0.75; p < 0.05 by Mann-Whitney U test). CONCLUSIONS: The induction donor inflammatory state before harvest leading to increased cardiac ICAM-1 expression promotes reperfusion injury and chronic graft vascular disease after transplantation in this rodent heterotopic heart model. PMID- 9354519 TI - Management strategy for simultaneous carotid endarterectomy and coronary revascularization. AB - BACKGROUND: The occurrence of significant carotid artery disease in patients requiring coronary artery bypass grafting (CABG) results in a dilemma regarding the best surgical management. Our philosophy has been to perform simultaneous carotid endarterectomy and CABG. We reviewed the efficacy of this therapy in patients treated at a large community-based hospital. METHODS: During a 6-year period, from 1990 to 1996, 88 patients underwent simultaneous carotid endarterectomy and CABG. All patients underwent preoperative four-vessel arch arteriography and standard coronary angiography. The principal indications for combined procedures were the need for CABG and (1) symptomatic carotid artery disease; (2) internal carotid artery stenosis of 80% or more, with or without contralateral disease; or (3) an ulcerated, unstable internal carotid artery lesion, regardless of degree of stenosis. The average patient age was 68 years, and there was a 3:1 male-to-female predominance. All procedures were performed with the patients under general anesthesia. The carotid endarterectomy was performed first, and an intraluminal shunt was used in all patients. RESULTS: The average degree of stenosis on the operated side was 86.2%. An average of 3.6 coronary bypasses per patient were performed. Morbidity included four strokes (4.5%). There were no perioperative myocardial infarctions. There were three hospital deaths (3.4%). The combined permanent stroke and mortality rate was 6.8%. Univariate predictors of stroke were an elevated serum creatinine level, a pulmonary complication, and left main coronary artery disease. Univariate predictors of hospital death were stroke, an elevated serum creatinine level, peripheral vascular disease, and left main coronary artery disease. Multivariate predictors of a prolonged hospitalization were stroke, an elevated serum creatinine level, and a pulmonary complication. Eighty-five patients (96.6%) were discharged and alive at 30 days. CONCLUSIONS: In the context of the indications we used to select patients for simultaneous carotid endarterectomy and CABG, the combined permanent stroke and mortality rate was less than 7%. Our management strategy identified patients that were at increased surgical risk as a result of advanced carotid and coronary artery disease. In our practice, simultaneous carotid endarterectomy and CABG is the preferred surgical approach for these high risk patients and results in a low in-hospital morbidity and mortality using a single anesthetic and hospitalization. PMID- 9354520 TI - Cardiac allograft vasculopathy is abrogated by anti-CD8 monoclonal antibody therapy. AB - BACKGROUND: Cardiac allograft vasculopathy, a diffuse and accelerated form of arteriosclerosis, is a major cause of graft loss or heart transplant recipient death after the first transplant year. This study examined the effects of depleting host CD8 + T lymphocytes on the development of cardiac allograft vasculopathy in miniature swine. METHODS: Cardiac allografts were heterotopically transplanted across a major histocompatibility complex class I barrier in partially inbred miniature swine and monitored for rejection by serial biopsies, electrocardiograms, and echocardiograms. Four control animals received cyclosporine on postoperative days 0 to 11. Another four miniswine were given 14.5 mg/kg of 76-2-11 (a mouse anti-swine CD8 monoclonal antibody) on postoperative day 0, in addition to a 12-day course of cyclosporine. Host CD8+ T cells and circulating 76-2-11 monoclonal antibodies were monitored by flow cytometry. RESULTS: As compared with cyclosporine-treated control animals, swine receiving 76-2-11 demonstrated near-complete depletion of peripheral CD8+ T cells by postoperative day 2, which persisted for 14 to 18 days. Mean allograft survival of the antibody-treated group and the control group was not statistically different (33 days versus 39 days, respectively) and both groups demonstrated severe interstitial rejection at necropsy. Control animals demonstrated florid intimal thickening of large and small arteries at necropsy. However, swine treated with 76-2-11 showed no intimal proliferation. CONCLUSIONS: Depletion of host CD8+ T cells prevents or delays the development of intimal proliferation in miniature swine. CD8+ lymphocytes play an important role in the early development of cardiac allograft vasculopathy in large animals. PMID- 9354521 TI - Distortions of the mitral valve in acute ischemic mitral regurgitation. AB - BACKGROUND: In the absence of papillary muscle rupture, the precise deformations that cause acute postinfarction mitral valve regurgitation are not understood and impair reparative efforts. METHODS: In 6 Dorsett hybrid sheep, sonomicrometry transducers were placed around the mitral annulus (n = 6) and at the tips and bases of both papillary muscles (n = 4). Later, specific circumflex coronary arteries were occluded to infarct approximately 32% of the posterior left ventricle and produce acute 2 to 3+ mitral regurgitation. Before and after infarction, distance measurements between sonomicrometry transducers produced three-dimensional coordinates of each transducer every 5 ms. RESULTS: After infarction, the annulus dilated asymmetrically orthogonal to the line of leaflet coaptation, but the annular area increased only 9.2% +/- 6.3% (p = 0.02). At end systole, posterior papillary muscle length increased 2.3 +/- 0.9 mm (p = 0.005); the posterior papillary muscle tip moved closer to the annular plane and centroid, and the anterior papillary muscle tip moved away. CONCLUSIONS: Small deformations in mitral valvular spatial geometry after large posterior infarctions are sufficient to produce moderate to severe mitral regurgitation. The most important changes are asymmetric annular dilatation, prolapse of leaflet tissue tethered by the posterior papillary muscle, and restriction of leaflet tissue attached to the anterior papillary muscle. PMID- 9354522 TI - Aortic root replacement: results using the St. Jude Medical/Hemashield composite graft. AB - BACKGROUND: Aortic root replacement remains a formidable operation. Although perioperative mortality has declined steadily, there is no consensus regarding the preferred method of reconstruction or type of composite to be used. We present our last 2 years' experience with aortic root replacement using the St. Jude Medical/Hemashield composite valve conduit. METHODS: A retrospective review of 52 consecutive patients undergoing aortic root replacement from February 1994 through October 1996 is presented. Both the open/exclusion and Cabrol methods of reconstruction were used. RESULTS: Thirty-one percent of the patients had undergone previous procedures of the aortic root. Thirty-seven percent required aortic arch replacement and 35% required concomitant cardiac or vascular procedures. Perioperative morbidity was low, as was perioperative mortality (3.8%). Both of the deaths that occurred were related to complications with the management of remaining thoracoabdominal aneurysms. CONCLUSIONS: Using meticulous surgical technique and the St. Jude Medical/Hemashield composite valve conduit, one can expect low mortality and complication rates for complex aortic root reconstruction. PMID- 9354524 TI - Surgical anatomy of the internal thoracic artery. AB - BACKGROUND: The internal thoracic artery (ITA) has become increasingly important in coronary artery bypass grafting due to the excellent long-term results. This study reviews its anatomic characteristics. METHODS: The ITAs of 100 cadavers were examined and their origin, relation to the phrenic nerve, presence of lateral costal branch; origin of pericardiacophrenic arteries, length, level and type of ITA termination, relation with the transverse muscle of thorax, collateral parietal branches, and distance between the ITA and sternal margins were studied. RESULTS: The ITA was present in all cases, originating directly from the subclavian artery or from a common trunk with other arteries. Its length was 20.4 cm on average, and the most frequent level of termination was at the sixth intercostal space, existing as a bifurcation in 93% and as a trifurcation in 7%. The pericardiacophrenic artery originated from the ITA in 89%. The lateral costal branch was present in 15% of the cases. The ITA was covered by the transverse muscle of the thorax for 7.5 cm (average) and was crossed anteriorly by the phrenic nerve in 70.0%. CONCLUSIONS: Information provided by this study may contribute to knowledge of its anatomic characteristics and in turn help prevent complications in ITA dissections. PMID- 9354523 TI - Catheter-assisted totally thoracoscopic coronary artery bypass grafting: a feasibility study. AB - BACKGROUND: The purpose of this study is to examine the feasibility of performing totally thoracoscopic internal mammary-to-coronary artery bypass grafting with the assistance of radiologically guided catheter intervention. METHODS: Fourteen dogs were subjected to mobilization of the internal mammary artery and anastomosis of it to the left anterior descending coronary artery over an angiographic catheter inserted into the internal mammary artery under fluoroscopy. The anastomosis was completed over the catheter using sutures and the application of fibrin glue. Eight animals underwent the anastomosis after their sacrifice. The other 6 animals were put on closed chest cardiopulmonary bypass and had their anastomosis done after intraaortic balloon occlusion and cardioplegic arrest of the heart. All animals had an angiographic and pathologic examination at the completion of the anastomosis. RESULTS: Anastomosis was completed in all dogs. Three anastomoses leaked and two were noted to be stenosed at completion of the anastomosis. One leak was sealed by application of fibrin glue. Both stenotic anastomoses were caused by suturing of the back wall when a short angiographic catheter could not be positioned across the anastomosis. CONCLUSIONS: Minimally invasive totally thoracoscopic mammary-to-coronary artery bypass grafting with catheter assistance is feasible. Technical improvement and appropriate instrumentation are required to minimize anastomotic failure. PMID- 9354525 TI - Hemodynamic advantages of left atrial epinephrine administration in open heart operations. AB - BACKGROUND: It is often necessary to administer a catecholamine to patients who have undergone cardiac operations. However, there are some potential disadvantages to using the central venous circulation, a routine route for catecholamine infusion. The advantages of the left atrial infusion of epinephrine were investigated in 21 patients. METHODS: The first group received epinephrine through the central venous route (central venous group), and the second group received adrenaline through the left atrial route (left atrial group). Hemodynamic studies were performed in all patients before and after the infusions. Blood samples were also taken from the radial and pulmonary arteries to determine the epinephrine concentrations. RESULTS: The average pulmonary arterial pressure and pulmonary vascular resistance were higher in the central venous group, whereas higher cardiac indices and average blood pressures were noted in the left atrial group (p < 0.05). There was a statistically significant difference in the epinephrine concentrations in the pulmonary arterial and radial arterial samples between the two groups. CONCLUSIONS: We conclude that epinephrine infusion through the left atrial route is associated with greater hemodynamic advantages than infusion through the central venous route. PMID- 9354526 TI - ICU admission score for predicting morbidity and mortality risk after coronary artery bypass grafting. AB - BACKGROUND: This study was performed to develop an intensive care unit (ICU) admission risk score based on preoperative condition and intraoperative events. This score provides a tool with which to judge the effects of ICU quality of care on outcome. METHODS: Data were collected prospectively on 4,918 patients (study group n = 2,793 and a validation data set n = 2,125) undergoing coronary artery bypass grafting alone or combined with a valve or carotid procedure between January 1, 1993, and March 31, 1995. Data were analyzed by univariate and multiple logistic regression with the end points of hospital mortality and serious ICU morbidity (stroke, low cardiac output, myocardial infarction, prolonged ventilation, serious infection, renal failure, or death). RESULTS: Eight risk factors predicted hospital mortality at ICU admission, and these factors and five others predicted morbidity. A clinical score, weighted equally for morbidity and mortality, was developed. All models fit according to the Hosmer-Lemeshow goodness-of-fit test. This score applies equally well to patients undergoing isolated coronary artery bypass grafting. CONCLUSIONS: This model is complementary to our previously reported preoperative model, allowing the process of ICU care to be measured independent of the operative care. Sequential scoring also allows updated prognoses at different points in the continuum of care. PMID- 9354527 TI - Coronary artery bypass grafting in immune thrombocytopenic purpura. AB - BACKGROUND: Reports of patients with idiopathic thrombocytopenic purpura undergoing cardiac operations are scarce and no recommendations exist regarding their management. We report 3 patients with idiopathic thrombocytopenic purpura and severe coronary artery disease who underwent uncomplicated coronary bypass grafting. METHODS: The case history of each patient with idiopathic thrombocytopenic purpura who underwent coronary artery bypass grafting and the literature were reviewed. RESULTS: All 3 patients underwent uncomplicated coronary artery bypass grafting after preoperative treatment with intravenous immunoglobulin and intraoperative platelet transfusions if needed. Prophylactic splenectomy was not performed. There was no increased incidence of bleeding complications. CONCLUSIONS: Coronary artery bypass grafting can be safely performed in patients with idiopathic thrombocytopenic purpura using conventional conduits after pretreating with immunoglobulin G and avoiding splenectomy. PMID- 9354528 TI - Prevention of calcification in glutaraldehyde-treated porcine aortic and pulmonary valves. AB - BACKGROUND: The problem of calcification in porcine aortic (AVs) and pulmonary (PVs) valves and its relationship to glutaraldehyde (GA) is of current interest. We proposed an anticalcification treatment to develop noncalcifying porcine AVs and PVs. METHODS: Porcine AVs and PVs were cross-linked in GA. Partially degraded heparin was coupled to the GA-treated AVs and PVs through intermediate surface bound substrate containing amino groups. Control AVs and PVs were cross-linked in 0.625% GA but had no heparin coupling. All specimens were implanted subdermally in 3-week-old rats for 5 months for calcification studies. RESULTS: Control AVs (Ca, 233.69 +/- 42.61 mg/g) and PVs (Ca, 181.48 +/- 4.06 mg/g) were severely calcified. Coupling of partially degraded heparin revealed complete prevention of calcification in GA-treated AVs (Ca, 0.73 +/- 0.27) and PVs (Ca, 1.125 +/- 0.22 mg/g) implanted subcutaneously in weanling rats for 5 months. CONCLUSIONS: The proposed anticalcification treatment is effective in preventing calcification of GA-treated AVs and PVs implanted subcutaneously in weanling rats for 5 months. PMID- 9354529 TI - Surgical treatment of aortic arch aneurysms in profound hypothermia and circulatory arrest. AB - BACKGROUND: This study was undertaken to define the factors that influence mortality rate and neurologic outcome after repair of the aortic arch and various portions of the thoracic aorta in patients with profound hypothermia and circulatory arrest. METHODS: Between November 1986 and January 1996, 105 patients were treated surgically for aortic disease involving the transverse aortic arch. Profound hypothermic circulatory arrest and selective brachiocephalic perfusion was used in all patients. In 19 patients retrograde cerebral perfusion was instituted during the period of circulatory arrest. Independent predictors for 30 day mortality and permanent neurologic deficits were evaluated by multiple logistic regression. RESULTS: Thirty-day mortality for the entire group was 19% (20/105); 21.2% for urgent versus 15.4% for elective cases, respectively. Statistical analysis showed that age is the most important factor that significantly influences mortality rate (p < 0.0145) and neurologic outcome (p < 0.006). Variables such as circulatory arrest time (p < 0.24), previous cardiac or aortic operations (p < 0.19), and sex (p < 0.55) failed to show any influence on mortality rate. Permanent neurologic deficits were diagnosed in 12.9% (11/85) of the patients. CONCLUSIONS: The incidence of permanent neurologic dysfunction as well as the mortality rate are predominantly related to the age of the patient. In this patient group, statistical analysis failed to show a direct correlation between duration of circulatory interruption and neurologic outcome. PMID- 9354530 TI - Late results of patch repair of coarctation of the aorta in adults using autogenous arterial wall. AB - BACKGROUND: Interposition grafting or patch repair of adult coarctations of the aorta are the standard methods of surgical treatment. Both involve use of prosthetic material, and patch repair using prosthetic material may lead to aneurysm formation in the long term. METHODS: Four patients aged 17 to 29 years had been investigated for systemic hypertension and had coarctation of the aorta diagnosed on cardiac catheterization. Between March and November 1984, all 4 underwent a corrective operation. The lesions were widely incised and a broad patch of ipsilateral mammary or Abbott's artery was fashioned across the narrowing. The arteries had been enlarged in diameter because of prolonged exposure to high blood pressure as collateral vessels, although none was intrinsically diseased. RESULTS: After 12 years of follow-up, only 1 patient remains on antihypertensive therapy. Spiral computed tomographic reconstructions revealed only very mild residual stenosis in 1 patient, confirmed by subsequent aortography. CONCLUSIONS: In adult patients with coarctation of the aorta, the use of the enlarged internal mammary artery as a patch graft is a simple, quick procedure, which may give lasting relief of obstruction. Spiral computed tomographic scanning is an ideal noninvasive method of follow-up. PMID- 9354531 TI - Effect of flush-perfusion on vascular endothelial and smooth muscle function. AB - BACKGROUND: The aim of this study was to investigate how much perfusion pressure an artery can tolerate without significant loss of endothelium-dependent relaxation (EDR) and vascular contractility. METHODS: The abdominal aortas of 396 Sprague-Dawley rats were used. One hundred twenty aortas were flush-perfused for 1 or 5 minutes with cold St. Thomas' Hospital cardioplegic (STHC) solution or with the same solution but modified by the addition of 3.5% dextran 40. Three perfusion pressures were tested: 50, 100, and 150 mm Hg. Two hundred eighty vessels were subjected to pressures of 50, 150, or 300 mm Hg using saline or STHC solution at 22 degrees C or STHC solution at 4 degrees C, for 10 or 60 seconds. The vessels were investigated in organ baths. Contractility was tested with the thromboxane analogue U-46619, acetylcholine was used to investigate EDR, and papaverine to elicit endothelium-independent relaxation. RESULTS: Flush-perfusion with cold STHC solution for 5 minutes at a perfusion pressure of 50 or 100 mm Hg affected neither contractility nor EDR. Vessels exposed to a flush-perfusion pressure of 150 mm Hg for 1 or 5 minutes lost 39% (p < 0.001) and 53% (p < 0.001) of their contractility, respectively. Flush-perfusion at 150 mm Hg for 1 minute did not affect EDR, whereas 5 minutes' perfusion caused a reduction of 7% (p < 0.05). A repetition of these experiments using STHC solution with 3.5% dextran 40 added gave no significantly different results. The impairment in contractility and EDR seen after perfusion at 150 mm Hg for 5 minutes disappeared after transplantation and reperfusion for 7 days. The vessels could be distended with saline or STHC solution at a pressure of 150 mm Hg without affecting contractility at 22 degrees C. At 4 degrees C, however, this pressure was harmful to contractility. Distention at a pressure of 300 mm Hg almost abolished contractility and 7 days after transplantation there had not yet been any recovery of contractility, but 30 days after transplantation the grafts had regained their normal contractility. CONCLUSIONS: Cold STHC solution, with or without dextran 40, can be used with a perfusion pressure of 100 but not 150 mm Hg without impairing EDR or vascular smooth muscle function. PMID- 9354532 TI - Immediate-early gene expression in ovine brain after hypothermic circulatory arrest: effects of aptiganel. AB - BACKGROUND: Altered gene expression occurs in the brain after global ischemia. We have developed a model to examine the effects of cardiopulmonary bypass and hypothermic circulatory arrest (HCA) on the induction of the immediate-early gene c-fos in the brains of neonatal lambs. We then tested the effects of the noncompetitive N-methyl-D-aspartate antagonist, aptiganel hydrochloride (Cerestat), on c-fos expression and neuronal injury. METHODS: Neonatal lambs (weight, 4 to 6 kg) anesthetized with isoflurane were supported by cardiopulmonary bypass, subjected to 90 or 120 minutes of HCA at 15 degrees C, and rewarmed on bypass to 38 degrees C. One hour after cardiopulmonary bypass was terminated, the brains were perfusion fixed and removed for in situ hybridization and immunohistochemical analysis. Some animals survived 3 days before their brains were removed to examine for neuronal necrosis. One group of lambs (n = 20) received aptiganel (2.5 mg/kg). A second group (n = 25) received saline vehicle only. RESULTS: Increasing duration of HCA induced a corresponding increase in c fos messenger RNA expression throughout the hippocampal formation and cortex. However, Fos protein synthesis peaked after 90 minutes of HCA and decreased significantly (p < 0.01) after 120 minutes of HCA. Aptiganel administration caused a significant decrease in (p < 0.001) c-fos messenger RNA expression and Fos protein synthesis after 90 minutes of HCA and preserved Fos protein synthesis after 120 minutes of HCA. Neuronal necrosis was observed in the brains of vehicle treated lambs after 120 minutes of HCA but was significantly decreased (p < 0.05) in the lambs given aptiganel. CONCLUSIONS: These experiments indicate that the transcriptional processes of immediate-early genes remain intact, whereas translational processes are impaired after prolonged HCA. The inability to synthesize Fos proteins after 120 minutes of HCA was associated with neuronal degeneration. Aptiganel preserved translational processes and caused a significant improvement in the neurologic outcome. PMID- 9354533 TI - Late survival after valve operation in patients with left ventricular dysfunction. AB - BACKGROUND: Left ventricular dysfunction is a predictor of hospital mortality after cardiac valve operation. We evaluated late survival in a large cohort of these patients. METHODS: From 1980 to 1993, 257 patients with a preoperative ejection fraction of 0.40 or less underwent aortic (n = 177), mitral (n = 72), or combined (n = 8) valve operation, with or without concomitant coronary artery bypass grafting. RESULTS: Hospital mortality was 12.5%. Follow-up was 98% complete. Logistic regression analysis showed that an ejection fraction of less than 0.30, mitral regurgitation, concomitant coronary artery bypass grafting, emergency operation, and reoperation were independent correlates of hospital mortality (all at p < 0.05). Kaplan-Meier survival curves of the 220 hospital survivors showed a 65% 5-year survival. Multivariate analysis revealed preoperative use of diuretics, male sex, reoperation, age exceeding 60 years, and aortic regurgitation to be independent predictors of poor late outcome (all at p < 0.05). CONCLUSIONS: The liability of left ventricular dysfunction with regard to diminished long-term survival is not completely reversed by valve operation. If operation is not performed before left ventricular dysfunction develops, postoperative medical treatment of these dilated, remodeled ventricles should be considered. PMID- 9354534 TI - In vivo morphology of woven, collagen-sealed Dacron prostheses in the thoracic aorta. AB - BACKGROUND: Long-term changes in knitted Dacron grafts inserted into the infrarenal aorta have been addressed by a number of studies indicating their potential for postoperative dilatation. In contrast, the behavior of woven, collagen-presealed, double-velour Dacron grafts used to replace the thoracic aorta is not known. METHODS: Forty-five patients were examined at a mean of 32.4 +/- 14.8 months after insertion of woven, collagen-coated, Dacron double-velour prostheses (Meadox woven with Hemashield, Meadox, Oakland, NJ) in the thoracic position under highly standardized conditions using spiral computed tomography. RESULTS: Compared with a manufactured diameter of 26 mm, all grafts showed an increase of 1 to 5 mm (mean, 3.0 +/- 1.2 mm [11.6% +/- 4.4%]; p < 0.0001) with greater enlargement of the ascending than of the descending aortic portions (p = not significant). A further statistically significant progressive dilatation failed to occur. Degenerative changes, including false aneurysm formation, could be excluded. CONCLUSIONS: Woven, collagen-coated Dacron prostheses are considered a safe replacement material for the thoracic aorta. PMID- 9354535 TI - Gradual reperfusion reduces infarct size and endothelial injury but augments neutrophil accumulation. AB - BACKGROUND: Reperfusion causes injury to the coronary artery endothelium primarily by neutrophil-mediated mechanisms. However, factors other than neutrophils may govern the extent of myocardial necrosis. This study tests the hypothesis that gradual initiation of reflow will reduce reperfusion injury and preserve postischemic endothelial function. METHODS: In 16 anesthetized dogs, the left anterior descending artery was ligated for 60 minutes. In one group, reperfusion was initiated abruptly (abrupt, n = 8), whereas in the gradual reperfusion group (ramp, n = 8), flow was slowly initiated during the first 30 minutes of reperfusion. After reperfusion, coronary artery segments were isolated to assess postischemic endothelial function. RESULTS: Infarct size (area of necrosis/area at risk) was significantly reduced in the ramp group (28.2% +/- 2.0%) compared with abrupt (41.6% +/- 1.4%). Neutrophil accumulation (myeloperoxidase) in the area at risk was significantly greater in the ramp group compared with abrupt (8.0 +/- 1.3 versus 3.5 +/- 0.8 U/g tissue). In isolated postischemic left anterior descending arterial rings, the concentration of acetylcholine that elicited a response 50% of the maximum possible response was significantly greater in abrupt (-6.88 +/- 0.04 log [mol/L]) than ramp (-7.62 +/- 0.04 log [mol/L]) and control (-7.68 +/- 0.003 log [mol/L]), suggesting endothelial dysfunction. The concentration of A23187 that elicited a response 50% of the maximum possible response was similarly greater in abrupt (-7.24 +/- 0.03 log [mol/L]) versus ramp (-7.62 +/- 0.03 log [mol/L]) and control (-7.8 +/- 0.04 log [mol/L]). Smooth muscle dysfunction (response to sodium nitrite) also occurred in the abrupt rings. CONCLUSIONS: Gradual reperfusion of an ischemic area reduces infarct size and preserves endothelial function but paradoxically increases neutrophil accumulation within the area at risk. PMID- 9354536 TI - Aortic valve conservation in acute type A dissection. AB - BACKGROUND: We consider operative survival as the primary objective in acute type A dissection and believe that virtually all native aortic valves can be conserved. We sought to answer the question: "Does glue repair improve the long term stability of proximal aortic repair?" METHODS: We retrospectively studied 64 patients with an acute type A dissection, an ascending aortic tear, and aortic regurgitation operated on by the same surgeon between 1988 and 1996. Three had Marfan's syndrome and 2 had a bicuspid valve. The valves in all patients without Marfan's syndrome were repaired with gelatin-resorcinol-formol glue. The valve and root were reinvestigated by echocardiography. Some patients underwent nuclear magnetic resonance imaging. RESULTS: There were four hospital (6%) and three late deaths. Aortic root reoperation was required in 2 of the 60 survivors (3.3%) and operation on the distal aorta in 2. Root reoperations were required within 3 years. The remaining proximal repairs remained stable. CONCLUSIONS: The native aortic valve can be conserved in most patients, and glue repair is durable. Simple root repair is associated with a low operative mortality. PMID- 9354537 TI - Prevention of supraventricular tachyarrhythmias after open heart operation by low dose sotalol: a prospective, double-blind, randomized, placebo-controlled study. AB - BACKGROUND: The aim of this prospective, double-blind, placebo-controlled trial was to assess the preventive effect and safety of low-dose sotalol after heart operation. METHODS: Two hundred fifty-five consecutive patients referred for elective coronary artery bypass grafting (n = 220) or aortic valve operation (n = 35) were randomized to receive either 80 mg of sotalol twice daily (n = 126) or matching placebo (n = 129) for 3 months, with the first dose given 2 hours before operation. RESULTS: There were no significant baseline differences between the groups. Overall, supraventricular tachyarrhythmias occurred in 36% of patients (82% atrial fibrillation). Hospital stay was 11.6 +/- 5 days in patients with supraventricular arrhythmias, versus 9.5 +/- 2.4 days in patients without it (p < 0.0001). Low-dose sotalol reduced the rate of supraventricular arrhythmias from 46% (placebo) to 26% (sotalol; p = 0.0012), or by 43%. On the fourth postoperative day, heart rate was lower in the sotalol group (74 +/- 12 beats/min versus 85 +/- 15 beats/min; p < 0.0001) but the QT interval corrected for the heart rate was not prolonged (sotalol group, 0.44 +/- 0.03 second; placebo group, 0.43 +/- 0.03 second; p = not significant). Study medication had to be discontinued because of side effects in 5.6% of sotalol and 3.9% of placebo patients (p = not significant), with one possible proarrhythmic event occurring in a patient receiving sotalol. CONCLUSIONS: Because more than 90% of supraventricular arrhythmic episodes occurred within 9 days after operation and 70% of all possibly sotalol related side effects occurred after day 9, the findings in this study imply that prophylactic treatment with sotalol may be limited to the first 9 postoperative days. PMID- 9354538 TI - Factors influencing HLA sensitization in implantable LVAD recipients. AB - BACKGROUND: Patients bridged to transplantation (TX) with the implantable left ventricular assist device (LVAD) may be at increased risk for the development of panel-reactive antibodies (PRA) during support. METHODS: To investigate that, we evaluated 60 patients who received the HeartMate LVAD at our institution, of whom 53 had PRA results available for analysis. T lymphocyte PRA levels were examined before LVAD, at the peak PRA level during LVAD support (PEAK), and just before TX. A PRA level more than 10% was considered indicative of sensitization against HLA antigens. RESULTS: The only factor that had a significant effect on PRA levels before LVAD was patient's sex (1.3% for men versus 7.4% for women; p = 0.005). During LVAD support, peak PRA levels increased significantly and the sex associated differences were no longer evident (33.3% men, 34.3% women; not significant). At the time of TX, PRAs decreased to 10.9% (men) and 7.0% (women) (not significant). We examined the influence of blood products received before TX on PRA levels. Patients who received less than the median number of total units (median). When examined by the type of blood product, only the number of platelet transfusions significantly increased the peak PRA (median: 46.9%; p = 0.03). Patients who received blood that was leukocyte depleted tended to have lower TX PRA levels (2.9%) compared with those who did not (13.9%, p = 0.18). Forty-two patients were successfully bridged to TX, with three early and two late deaths after TX. Whereas 39 patients received transplants without intervention, 3 were treated by plasmapheresis with a 77% reduction in their HLA antibody levels at TX as measured by flow cytometry. CONCLUSIONS: Patients with the implantable LVAD are at significant risk for the development of anti-HLA antibodies during support. Although this sensitization is often transient, intervention using plasmapheresis may be useful for some patients. PMID- 9354539 TI - Modified Norwood operation for single left ventricle and ventriculoarterial discordance: an improved surgical technique. AB - BACKGROUND: Patients with univentricular hearts and ventriculoarterial discordance with potentially obstructed systemic blood flow continue to pose difficult management problems. The goals of neonatal palliative operations are to control pulmonary blood flow while avoiding pulmonary artery distortion, to relieve systemic outflow tract obstruction, and to avoid heart block. METHODS: Between January 1987 and December 1996, 38 patients with either tricuspid atresia or a double-inlet left ventricle and ventriculoarterial discordance underwent a modified Norwood procedure. Their mean age was 15 days, and their mean weight was 3.4 kg. Aortic arch anomalies were present in 92% of the patients. Morbidity and mortality statistics, intraoperative data, and postoperative echocardiograms were reviewed. RESULTS: There were 3 early deaths (7.8%) and 5 late deaths (13.1%). The actuarial survival rates at 1 month, 1 year, and 5 years were 89%, 82%, and 71%, respectively. Follow-up was complete in all children at a mean interval of 30 +/- 9 months. None of the patients had significant neoaortic valve insufficiency, and 1 patient required therapy for residual aortic arch obstruction. Nine patients (30% of the survivors) have undergone the hemi-Fontan procedure, and 18 patients (60%) successfully have undergone the Fontan procedure. CONCLUSIONS: In this patient population, we recommend the modified Norwood procedure as the neonatal palliative treatment of choice. It can be performed with acceptable early morbidity and mortality, and it improves suitability for the Fontan procedure. It reliably relieves all levels of systemic outflow tract obstruction, controls pulmonary blood flow, and avoids heart block. PMID- 9354540 TI - Continuous versus intermittent furosemide infusion in critically ill infants after open heart operations. AB - BACKGROUND: Use of intravenous furosemide is generally avoided in critically ill neonates and infants soon after open heart operations to prevent fluctuations in intravascular volume and resulting circulatory instability. METHODS: To assess and compare the safety and efficacy of continuous versus intermittent intravenous furosemide, we undertook a prospective, randomized trial in 26 consecutive patients less than 6 months of age. Inclusion criteria were presence of low output syndrome requiring inotropic support (24/26 patients) or pulmonary hypertension requiring vasodilator therapy (10/26 patients) within 6 hours of discontinuation of cardiopulmonary bypass. Eleven patients received 0.1 mg x kg( 1) x h(-1) continuous intravenous furosemide (group 1) and 15 received 1 mg/kg bolus every 4 hours (group 2) for 24 hours. Mean age (3.7 +/- 3.4 versus 1.8 +/- 2.5 months) and weight (4.6 +/- 2.1 versus 4.3 +/- 1.7 kg) were comparable. RESULTS: Group 2 infants showed slightly greater absolute urinary output (2.5 +/- 1.1 mL/kg per hour versus 3.3 +/- 1.1 mL/kg per hour, p = 0.05). However, urinary output per dose of drug was significantly larger in group 1 infants (1.0 +/- 0.4 versus 0.5 +/- 0.2 mL x kg(-1) x h(-1); p = 0.002) with lesser fluctuations (variance, 1.9 +/- 1.6 versus 3.8 +/- 2.1; p = 0.02) and fluid replacement needs (20.6 +/- 3.8 versus 51.8 +/- 14.4; p = 0.001). Electrolyte replacement requirements were similar. A trend toward greater hemodynamic instability in group 2 patients (heart rate variance 88.4 +/- 79.8 versus 128.3 +/- 82.7; p = 0.09; central venous pressure variance 2.8 +/- 1.90 versus 4.1 +/- 3.7; p = 0.07; mixed venous oxygen saturation variance, 32.3 +/- 27.6 versus 45.7 +/- 20.4; p = 0.06) was noted. All patients who completed the study protocol survived operation and were discharged home. CONCLUSIONS: We conclude that (1) commonly used doses of both intermittent and continuous intravenous furosemide infusion can be safely administered to critically ill neonates and infants as early as 6 hours after operation, (2) continuous infusion yields an almost comparable urinary output with a much lower dose of furosemide, and (3) intermittent administration is associated with greater fluctuations in urinary output and greater needs for fluid replacement therapy. PMID- 9354541 TI - Cardiac operations in children with Marfan's syndrome: indications and results. AB - BACKGROUND: The development of new screening techniques for the early detection of Marfan's syndrome has prompted evaluation of the results of cardiac operations in children with this syndrome. The purpose of this study was to determine the surgical indications, operative results, and need for reoperation in children with Marfan's syndrome. METHODS: From 1980 to 1996, 245 patients underwent cardiac operations for complications of Marfan's syndrome; 26 (11%) were less than 18 years of age. The mean age at the time of operation was 10.3 +/- 1 years (range, 8 months to 17 years); 18 of the patients were male. Indications for operation were aortic root dilatation (15 patients), mitral regurgitation (4 patients), aortic root dilatation and mitral regurgitation (6 patients), and aortic arch aneurysm (1 patient). Operations included aortic root replacement (15 patients), aortic root replacement and mitral repair (5 patients), aortic root replacement and mitral replacement (1 patient), mitral repair (3 patients), mitral replacement (1 patient), and arch aneurysm repair (1 patient). The mean aortic root diameter in patients undergoing aortic root replacement was 6.2 +/- 0.2 cm. Only 1 patient underwent ascending aortic dissection. RESULTS. There were no operative deaths. At a mean follow-up of 67.1 +/- 10.2 months, 8 patients required a second cardiac procedure (41% +/- 17% 10-year freedom from reoperation). Indications for further operations were distal aortic pathology (3 patients), aortic root dilatation after initial mitral operation (3 patients), failed mitral repair (1 patient), and homograft degeneration (1 patient). Risk factors for a second cardiac procedure were age less than 10 years at the time of the first operation (p < 0.003) and mitral regurgitation (p < 0.04). Overall, 25 (96%) of 26 patients have undergone aortic root replacement and 11 (42%) patients have undergone a mitral procedure. There have been 4 late deaths, all of presumed cardiac origin. The 10-year survival rate is 79% +/- 10%. All surviving patients are in New York Heart Association functional class I or II. CONCLUSIONS: We conclude that (1) aortic root dilatation is the most common surgical indication in children with Marfan's syndrome, (2) mitral regurgitation is the second most common indication, (3) aortic dissection is unusual in children with Marfan's syndrome, and (4) careful follow-up is necessary, particularly in younger children, because more than half of all children with Marfan's syndrome require repeated cardiac operations within 10 years. PMID- 9354542 TI - Surgical strategy for doubly committed subarterial ventricular septal defect with aortic cusp prolapse. AB - BACKGROUND: Many surgeons recommend early repair of doubly committed subarterial ventricular septal defect regardless of the clinical symptoms. We reviewed our patients of this anomaly with aortic cusp prolapse to justify this strategy. METHODS: We reviewed the preoperative and postoperative records of 27 patients with doubly committed subarterial ventricular septal defect and aortic cusp prolapse. The patients' ages ranged from 2 months to 11 years (median, 4.6 years). RESULTS: During the preoperative observation period, aortic regurgitation (AR) developed in 65% of the patients. In the 8 patients without AR before the operation, AR did not develop after the operation, whereas AR persisted in 12 (63%) of the 19 patients with preoperative AR. To identify the risk factors for persistent AR after the operation we analyzed the data for the patients with preoperative AR in the persistent AR group (n = 12) and eliminated AR group (n = 7) and found a longer period from the onset of AR to the operation in the persistent AR group (32.1 +/- 10.1 versus 5.6 +/- 1.9 months; p = 0.014). During the follow-up period 10 of the 17 patients with mild AR before the operation showed persistent AR in the postoperative period, but it did not progress. CONCLUSIONS: We conclude that early surgical repair with a minimum observation period is essential for prevention of residual AR. Even if a tiny AR is detected preoperatively, the patient should be surgically treated immediately. PMID- 9354543 TI - Surgical repair of patients with tetralogy of Fallot and unilateral absence of pulmonary artery. AB - BACKGROUND; Patients with tetralogy of Fallot and unilateral absence of pulmonary artery are a high-risk group for whom there is no consensus on the correct approach to medical management. The purpose of this report is to review a 29-year experience in the treatment of those patients. METHODS: Between May 1966 and February 1995, 2,511 patients underwent correction of tetralogy of Fallot in our department, 24 of those patients with unilateral absence of pulmonary artery (20 had absence of the left pulmonary artery, 4 had absence of the right pulmonary artery). Valved conduits were used in 9 patients, right ventricular patches were used in 4 patients, and transannular patches with a monocusp that was made of the patient's pericardium were used in 11 patients. RESULTS: There were two operative deaths; both were in patients with hypoplasia of the left ventricle. All survivors had good early and late results. CONCLUSIONS: A right ventricular patch should be used in patients with tetralogy of Fallot and infundibular stenosis; a transannular patch with a monocusp should be used in patients with tetralogy of Fallot and stenosis of the left or right pulmonary artery's origin as well as the pulmonary trunk. A homograft valved conduit is suitable for patients with anomalous coronary artery or pulmonary atresia. PMID- 9354544 TI - Left ventricular reduction operation in ischemic cardiomyopathy: a note of caution. AB - Surgical reduction has been proposed to reduce left ventricular wall stress and improve geometry in patients with left ventricular failure. We describe 2 cases of direct surgical reduction in ischemic cardiomyopathy. Although left ventricular function improved in both, late ventricular dysrhythmia negated the result. PMID- 9354545 TI - Simultaneous use of an implanted defibrillator and ventricular assist device. AB - A left ventricular assist device was placed as a bridge to cardiac transplantation in a 51-year-old man with cardiogenic shock. Placement of the left ventricular assist device occurred 5 years after implantation of an implantable cardioverter/defibrillator. The implantable cardioverter/defibrillator discharged appropriately during ventricular assist device support to terminate episodes of sustained ventricular tachycardia without causing malfunction of the ventricular assist device. PMID- 9354546 TI - Onlay patch repair of tracheobronchial rupture. AB - Two cases are reported of tracheobronchial repair in which a posteriorly based intercostal muscle flap was incorporated into the membranous portion of the airway to increase the diameter of the reconstruction or to relieve tension in the suture lines. This technique permits repair of a small left main bronchus without compromise to the lumen and tension-free repair of the membranous trachea. PMID- 9354547 TI - Cervicomediastinal hibernoma. AB - Hibernoma is a benign soft-tissue tumor, derived from the brown fat, that often presents as a painless, slow-growing mass. About 100 cases of hibernomas have been reported in the world literature. Seven cases of intrathoracic hibernoma are reported, of which only 1 was located in the mediastinal region. That tumor was an intramediastinal hibernoma with a cervicomediastinal location, which was excised through an extended left supraclavicular incision without the necessity to perform a sternotomy. No recurrence was evident after 18 months. PMID- 9354548 TI - Lymphoepithelioma-like carcinoma of the lung. AB - Primary lymphoepithelioma-like carcinoma of the lung is rare; only 26 case reports have been identified in the literature. The present report presents a case of a 67-year-old white man with a T1 N1 M0 lymphoepithelioma-like carcinoma of the lung. He presented with severe arthritic complaints that resolved after resection of the tumor. The majority of these tumors have occurred in Asian patients who have shown evidence of previous exposure to the Epstein-Barr virus. PMID- 9354549 TI - Atrial replacement and tricuspid valve reconstruction after angiosarcoma resection. AB - A cardiac angiosarcoma was resected and successfully managed by replacement of the right atrium and bileaflet reconstruction of the tricuspid valve by conserving non-involved valvular tissue. Competency of the new valve was confirmed intraoperatively by transesophageal echocardiography and reconfirmed at discharge. Evaluation 3 months postoperatively revealed no evidence of valvular insufficiency or right heart failure. In selected patients, resection of extensive primary cardiac neoplasms may be possible without necessitating prosthetic valve replacement. PMID- 9354550 TI - Berry syndrome, a complex aortopulmonary malformation: one-stage repair in a neonate. AB - Successful one-stage repair of a Berry syndrome (interrupted aortic arch, distal aortopulmonary septal defect, right pulmonary artery branch originating from the ascending aorta, and intact ventricular septum) in the neonatal period has been reported in only 2 cases. We report the case of a newborn operated on with deep hypothermic arrest and isolated myocardial perfusion in whom the interrupted aortic arch was corrected by direct anastomosis between the ascending and descending aorta and the aortopulmonary septal defect was treated with reconstruction of the pulmonary trunk and right pulmonary artery, using a flap of aortic tissue. A native pericardial patch was used to reconstruct the ascending aorta. PMID- 9354551 TI - Mitral valve myxoma. AB - Cardiac myxomas arising from the mitral valve are extremely rare. We describe the case of an asymptomatic 49-year-old woman who was found to have a 3.6 x 4.0-cm myxoma originating from the atrial side of the anterior mitral leaflet. The lesion was successfully treated by surgical excision and mitral valve replacement. A review of the literature regarding this rare lesion is presented. PMID- 9354552 TI - Cardiac entrapment during minimally invasive aortic valve replacement. AB - Reduced exposure during minimally invasive valve operations poses new difficulties in intraoperative management. Transesophageal echocardiography improves intraoperative management. During a minimally invasive aortic valve replacement, we encountered unexpected hypotension due to mechanical compression of the right ventricle against the sternum. Transesophageal echocardiography facilitated rapid diagnosis of this problem. Surgeons performing these procedures should be aware of this potential problem. PMID- 9354553 TI - Neurofibroma of the esophagus. AB - Benign esophageal tumors occur infrequently, with leiomyomas accounting for approximately 70% of cases. Benign neural tumors of the esophagus account for 200 cases reported in the literature and rarely require operative resection. The case of a 58-year-old woman with a 4-month history of progressive dysphagia and odynophagia is presented. A large intramural esophageal mass was resected through a right thoracotomy, and the esophagus was primarily repaired. Histologic examination revealed a neurofibroma. PMID- 9354554 TI - Unusual cause of pulmonary hypertension and congestive heart failure in a newborn. AB - Anomalous systemic arterial supply to a lobe of the lung is a rare cause of pulmonary hypertension and congestive heart failure in the newborn period. We report the presentation and successful treatment of a neonate with this unusual anatomy. Proper diagnosis required both echocardiography and aortography, and surgical resection of the involved lobe was curative. PMID- 9354555 TI - Single-incision and single-element array electrode to lower the defibrillation threshold. AB - Occasional patients have excessive defibrillation energy requirements despite appropriate transvenous defibrillation lead position and the use of biphasic shocks. A single-element subcutaneous array electrode was implanted in 2 patients with a high defibrillation threshold. The array electrode was implanted through the same infraclavicular incision that was used for implantation of the transvenous lead. The defibrillation threshold decreased from 30 J to 15 J and from 24 J to 9 J with the subcutaneous array electrode. PMID- 9354556 TI - Mitral valve repair in a patient with severe porcelain aorta. AB - We repaired the mitral valve in a patient with severe porcelain aorta. Significant mitral regurgitation developed in a 66-year-old woman with heavy calcification throughout the whole aorta. At operation, cardiopulmonary bypass was properly established by combined axillary and femoral arterial cannulations for sufficient systemic flow. Likewise, the combination of a superior mitral approach and profound hypothermic fibrillatory arrest in conjunction with low flow cardiopulmonary bypass allowed us to repair the mitral valve successfully. PMID- 9354557 TI - Ruptured left ventricular diverticulum in infancy. AB - Congenital diverticulum of the left ventricle is rare, and rupture of such a diverticulum is even more rare. We describe successful surgical repair of a ruptured left ventricular diverticulum in an 11-month-old infant. PMID- 9354558 TI - Endoscopic saphenous vein harvesting: minimally invasive video-assisted saphenectomy. AB - A technique of greater saphenous vein harvesting for coronary artery revascularization using an endoscopic approach is herein detailed. The saphenous vein is directly identified at the knee through a single incision. An endoscopic dissector is advanced proximally and distally along the course of the vein, ligating side-branches with clips. The vein is divided at the ends of dissection, dependent on patient anatomy, by either a counterincision, endoscopic clips, or ligation with an Endo-loop. PMID- 9354559 TI - Surgical clip for compression of the aortic wall during gluing. AB - An acrylic clip was developed to compress the dissected aortic wall during gluing. These small instruments have a concave-convex contact face adjusted to the most common graft size. There are several advantages to this method of treatment of acute aortic dissection. PMID- 9354560 TI - Aortic mismatch in heart transplantation: readaptation. AB - Great vessel mismatch between donor and recipient is very usual in heart transplantation. Different procedures have been used to manage this situation. A tailoring aortoplasty is described, as a technical alternative, in cases of considerable size incongruence between donor and recipient aortic diameters. PMID- 9354561 TI - Thoracoscopic fine-needle aspiration of solitary pulmonary nodules. AB - To determine the diagnostic efficacy of thoracoscopic fine-needle aspiration (FNA) of solitary pulmonary nodules suspicious for lung cancer, we performed intraoperative thoracoscopic FNA for diagnostic purposes in 8 consecutive patients with peripheral solitary pulmonary nodules suspicious for lung cancer. Thoracoscopic FNA yielded an accurate diagnosis in all cases. There were 5 cases of non-small cell lung carcinoma, 1 small cell lung carcinoma, 1 renal carcinoma metastasis, and 1 inflammatory nodule. Results of FNA were obtained in less than 10 minutes in 6 cases. Maximum time to diagnosis was 20 minutes. The surgical procedure was expedited in the 6 cases of lung cancer because lobectomy followed FNA rather than the performance of a diagnostic wedge resection. A minor hematoma after FNA was the single complication. Thoracoscopic FNA yielded a prompt and accurate diagnosis of peripheral solitary pulmonary nodules. Thoracoscopic FNA should be considered as an alternative to preoperative percutaneous FNA, which risks pneumothorax and patient discomfort. In cases of lung cancer, thoracoscopic FNA allows the surgeon to bypass a diagnostic wedge resection and to proceed with definitive lobectomy. PMID- 9354562 TI - Ventriculotomy repair during revascularization of intracavitary anterior descending coronary arteries. AB - Optimal revascularization of the rare variant anomolous intracavitary left anterior descending coronary artery requires, by definition, entrance into the right ventricular cavity. We present a simple method to repair the ventriculotomy without risk of obliterating the left anterior descending coronary artery, septal perforators, or diagonal branches. PMID- 9354563 TI - Implantation of the Toronto SPV stentless porcine bioprosthesis in dilated ascending aorta. AB - In the presence of severe dilatation of the ascending aorta the implantation of a Toronto SPV stentless bioprosthesis is compromised by the risk of postoperative central regurgitation. A modification of the implantation technique is described that restores the normal shape of the ascending aorta and thereby avoids the risk of dysfunction of the prosthesis. PMID- 9354564 TI - Cemil Topuzlu Pacha and his arterial suture technique. AB - Cemil Topuzlu Pacha (1868-1958) is known to be one of the most famous surgeons in Turkey through the early decades of the twentieth century. Being a talented and courageous surgeon, he performed many of the avoided operations of that time. He presented his vascular suture techniques at the International Medical Congress in Moscow in August 1897 and at the annual Congress of the Societe de Chirurgie de Paris in July 1904. He reported 2 cases of arterial tear during breast carcinoma resection and repair within the same session. He also reported the removal of a pen cover from the right main bronchus of a 7-year-old girl through a tracheotomy in 1903. He worked for 3 years with the famous French surgeon Jules Pean and became a preferred surgeon of the Ottoman Imperial family in Istanbul. He was admired for his scientific studies in international congresses and was one of the first Turkish surgeons who became a member of important European surgical associations. PMID- 9354566 TI - Thoracic Surgery Directors Association report 1995-1997. PMID- 9354565 TI - Angiogenesis in the pathobiology and treatment of vascular and malignant diseases. AB - Cardiovascular disease and cancer account for the majority of adult disease in the developed world. This review focuses on current concepts in the study of angiogenesis (new vessel formation) as related to these conditions and highlights the role of vascular endothelial growth factor. Developments in therapeutic angiogenesis have raised the possibility that pharmacologic or gene-directed interventions, based on the ability of vascular endothelial growth factor to promote new vessel formation, may soon gain clinical application for the treatment of occlusive vascular disease. Similarly, the future treatment of malignant disease is likely to involve antiangiogenic agents that, in preliminary animal work, have demonstrated an efficacy that is not limited by adverse affects. Aside from these potential applications, current investigations have enhanced our understanding of mechanisms involved in the development of atherosclerotic and malignant disease. PMID- 9354567 TI - A proper name for the internal mammary artery? PMID- 9354568 TI - An unusual type of total anomalous pulmonary venous connection. PMID- 9354569 TI - Stroke from heparin-coated circuits and reduced systemic anticoagulation. PMID- 9354570 TI - Internal thoracic artery spasm and normothermic papaverine. PMID- 9354571 TI - PTFE membranes, redo operations, and epicardial echocardiography. PMID- 9354572 TI - Maximal use of internal mammary artery. PMID- 9354573 TI - Effects of different methods of internal mammary artery harvesting. PMID- 9354574 TI - Low heparinization and heparin-bonded circuits. PMID- 9354575 TI - Rapid recovery in the elderly. PMID- 9354576 TI - Aortic valve repair in children. PMID- 9354577 TI - Noncardioplegic myocardial protection for CABG. PMID- 9354578 TI - Approach to severe blunt thoracic trauma and thoracoplasty. PMID- 9354579 TI - Cognitive outcome after CABG. PMID- 9354580 TI - Papillary fibroelastoma. PMID- 9354581 TI - Delayed sternal closure with sternal zipper. PMID- 9354582 TI - Traumatic subarachnoid-pleural fistula. PMID- 9354583 TI - Conservative treatment of tracheal lacerations secondary to endotracheal intubation. PMID- 9354584 TI - Putative mechanism through which N-cadherin-mediated cell contact maintains calcium homeostasis and thereby prevents ovarian cells from undergoing apoptosis. AB - To date most of the studies involving the maintenance of ovarian cell viability have focused on the endocrine, paracrine, and autocrine factors that inhibit these cells from undergoing programmed cell death or apoptosis. Recently, studies have demonstrated that cell contact also prevents ovarian cells from dying via an apoptotic mechanism. In this commentary, the role that homophilic binding of the cell adhesion molecule, N-cadherin, plays in maintaining ovarian cell viability is presented. These studies showed that N-cadherin homophilic binding (1) is part of the mechanism through which cell contact maintains cell viability, (2) results in the activation (i.e. tyrosine phosphorylation) of the fibroblast growth factor (FGF) receptor, and (3) prevents a sustained elevation in intracellular free calcium ([Ca2+]i) which triggers apoptosis. These studies also revealed that hepatocyte growth factor (HGF), also known as scatter factor (SF), disrupts cell contact, which leads to a sustained increase in [Ca2+]i levels and ultimately to cell death. Based on these studies, this commentary presents a putative mechanism that relates the cellular and molecular mechanism through which basic FGF, N cadherin, and HGF/SF interact to regulate [Ca2+]i levels and ultimately ovarian cell survival. PMID- 9354585 TI - Inhibition of murine macrophage nitric oxide synthase expression by a pivoxil prodrug of antiviral acyclic nucleotide analogue 9-(2 phosphonomethoxyethyl)adenine. AB - The effect of the acyclic nucleotide analogue, 9-(2-phosphonomethoxyethyl)adenine (PMEA, Adefovir), and its (bis)pivaloyloxymethyl ester (bis-POM-PMEA, Adefovir Dipivoxil) on in vitro nitric oxide (NO) production by murine peritoneal macrophages was investigated. Bis-POM-PMEA inhibited in a concentration-dependent manner the formation of NO generated by interferon-gamma and lipopolysaccharide, the IC50 being 15 microM. Suppressed transcription of mRNA for inducible NO synthase (EC 1.14.13.39) resulting in decreased synthesis of NO synthase protein was found. Parent compound PMEA was virtually ineffective. PMID- 9354586 TI - Mechanisms of the inhibition of human erythrocyte pyridoxal kinase by drugs. AB - The aim of this study was to investigate the interaction between drugs chosen for their clinical neurotoxicity or chemical structure and vitamin B6 metabolism. After a preliminary screening of drugs to determine their potential inhibitory effect on erythrocyte nonpurified pyridoxal kinase (PLK) (EC 2.7.1.35), additional investigations, including kinetic studies and detection of chemical reactivity between the inhibiting drugs and pyridoxal (PL) or pyridoxal-5' phosphate (PLP), using UV-visible spectrophotometry and mass analysis, were carried out to specify the mechanism of PLK inhibition. Depending on the results, the inhibiting drugs were divided into three groups. The first group included theophylline and progabide and inhibited PLK using either PL or pyridoxamine (PM) as substrate and thereby were true inhibitors. Moreover, they did not form covalent complexes with PL or PLP. The second group, which included cycloserine, dopamine, isoniazid, and thiamphenicol glycinate, inhibited PLK using PL, but not PM, as substrate. They were able to react with PL or PLP to form covalent complexes, and kinetic studies suggested that the observed PLK inhibition was due to these formed complexes. A third group, which consisted of levodopa, D penicillamine, and muzolimine, inhibited PLK using PL, but not PM, as substrate. They formed, with PL or PLP, chemical derivatives that probably had no inhibitory effect on PLK. These results and the clinical consequences of such interactions are discussed and compared with results of previous studies. PMID- 9354587 TI - The potent antioxidant activity of the vitamin K cycle in microsomal lipid peroxidation. AB - In the vitamin K cycle, vitamin K-hydroquinone, the active cofactor for gamma glutamylcarboxylase, is continuously regenerated. The successive pathways contain oxidation of the hydroquinone to the epoxide, followed by reduction to the quinone and reduction to the hydroquinone. Vitamin K-hydroquinone is a potent radical scavenging species (Mukai et al., J Biol Chem 267: 22277-22281, 1992). We tested the potential antioxidant activity of the vitamin K cycle in lipid peroxidation reactions (thiobarbituric acid reactive substances, TBARS) in rat liver microsomes. As prooxidant we used Fe2+/ascorbate, NADPH-Fe3+/ATP, and NADPH/CCl4. Vitamin K (< or = 50 microM) on its own did not influence the formation of TBARS. In combination with 1 mM dithiothreitol (DTT), the reductive cofactor for the microsomal enzyme vitamin K epoxide reductase, vitamin K suppressed lipid peroxidation with a concentration that blocked the maximal response by 50% (IC50) of ca. 0.2 microM. Vitamin K1 (phylloquinone) and vitamin K2 (menaquinone-4) were equally active. Warfarin (5 microM) and chloro-vitamin K (50 microM), inhibitors of vitamin K epoxide reductase and gamma glutamylcarboxylase, respectively, were able to completely abolish the antioxidant effect. Lipid peroxidation was inversely related to the amount of vitamin K hydroquinone in the reaction. Vitamin K epoxide reductase seemed sensitive to lipid peroxidation, with half of the activity being lost within 10 min during oxidation with NADPH/CCl4. The inactivation could be attenuated by antioxidants such as vitamin E, reduced glutathione, and menadione and also by a K vitamin in combination with DTT, but not by superoxide dismutase and catalase. The results show that the vitamin K cycle could act as a potent antioxidant, that the active species in all probability is vitamin K-hydroquinone, and that the primary reaction product is the semiquinone. The results also show that the reaction product is processed in the vitamin K cycle to regenerate vitamin K hydroquinone. PMID- 9354588 TI - Biochemical and antiproliferative properties of 4 [ar(alk)ylamino]pyridopyrimidines, a new chemical class of potent and specific epidermal growth factor receptor tyrosine kinase inhibitor. AB - The tyrosine kinase inhibitors PD 69896, 153717, and 158780, which belong to the chemical class 4-[ar(alk)ylamino]pyridopyrimidines, have been characterized with respect to enzymology, target specificity, and antiproliferative effects in tumor cells. These compounds were competitive inhibitors with respect to ATP against purified epidermal growth factor (EGF) receptor tyrosine kinase and inhibited EGF receptor autophosphorylation in A431 human epidermoid carcinoma with IC50 values of 2085, 110, and 13 nM, respectively. Onset of inhibition was immediate once cells were exposed to these compounds, whereas recovery of receptor autophosphorylation activity after the cells were washed free of the compound was dependent on inhibitory potency. Thus, full activity returned immediately after removal of PD 69896 but required 8 hr after exposure to PD 158780. PD 158780 was highly specific for the EGF receptor in Swiss 3T3 fibroblasts, inhibiting EGF dependent receptor autophosphorylation and thymidine incorporation at low nanomolar concentrations while requiring micromolar levels for platelet-derived growth factor- and basic fibroblast growth factor-dependent processes. PD 158780 inhibited heregulin-stimulated phosphorylation in the SK-BR-3 and MDA-MB-453 breast carcinomas with IC50 values of 49 and 52 nM, respectively, suggesting that the compound was active against other members of the EGF receptor family. The antiproliferative effects of this series of compounds against A431 cells correlated precisely with the inhibitory potency against EGF receptor autophosphorylation. PD 158780 reduced clone formation in soft agar of fibroblasts transformed by EGF, EGF receptor, or the neu oncogene but not ras or raf, further demonstrating its high degree of specificity. Finally, this compound was active against clone formation in several breast tumors having different expression patterns of the erbB family, indicating an anticancer utility in tumors expressing these receptors. PMID- 9354589 TI - Regulation of CYP4A1 and peroxisome proliferator-activated receptor alpha expression by interleukin-1beta, interleukin-6, and dexamethasone in cultured fetal rat hepatocytes. AB - The CYP4A1 isoenzyme induced in rodents by peroxisome proliferators is known to be repressed at a pretranslational level by interferon. Interleukin-1beta (IL 1beta) also reduces CYP4A1-related 12-laurate hydroxylase activity in cultured fetal rat hepatocytes after induction by clofibric acid. In this fetal hepatocyte model, IL-1beta and interleukin-6 (IL-6) were tested for their ability to reduce 12-laurate hydroxylase activity, CYP4A1 apoprotein content, and the CYP4A1 mRNA level. IL-1beta and IL-6 strongly diminished CYP4A1 activity and apoprotein and mRNA levels in a dose- and time-dependent manner. CYP4A1 expression is thus down regulated at a pretranslational level by these cytokines. As it has been shown that the peroxisome proliferator-activated receptor alpha (PPAR alpha) mediates the induction of the CYP4A1 gene by a peroxisome proliferator, the capacity of IL 1beta or IL-6 to modulate the PPAR alpha mRNA level was tested. It was found that IL-1beta and IL-6 both repress the induction of PPAR alpha expression exerted by the combined action of clofibric acid and dexamethasone. However, even at the highest concentration (10 ng/mL) tested for both cytokines, IL-1beta as well as IL-6 failed to abolish the induction of CYP4A1 by dexamethasone. The mechanism of the protective effect of the synthetic glucocorticoid on CYP4A1 repression by interleukins is discussed. PMID- 9354590 TI - Inhibition of proliferation and apoptosis of human and rat T lymphocytes by curcumin, a curry pigment. AB - Curcumin (diferuoylmethane), the yellow pigment in the rhizome of tumeric (Curcuma longa), an ingredient of curry spice, is known to exhibit a variety of pharmacological effects including antitumor, antiinflammatory, and antiinfectious activities. Although its precise mode of action remains elusive, curcumin has been shown to suppress the activity of the AP-1 transcription factor in cells stimulated to proliferate. In this study, we observed that curcumin (50 microM) inhibited proliferation of rat thymocytes stimulated with concanavalin A (Con A) as well as that of human Jurkat lymphoblastoid cells in the logarithmic growth phase. The pigment also inhibited apoptosis in dexamethasone-treated rat thymocytes and in UV-irradiated Jurkat cells as judged by DNA ladder formation, cellular morphological changes, and flow cytometry analysis. The inhibition of apoptosis by curcumin in rat thymocytes was accompanied by partial suppression of AP-1 activity. Complete suppression of AP-1 activity was observed in Con A treated, proliferating thymocytes. The capacity of curcumin to inhibit both cell growth and death strongly implies that these two biological processes share a common pathway at some point and that curcumin affects a common step, presumably involving a modulation of the AP-1 transcription factor. PMID- 9354591 TI - Promotion of HL-60 cell differentiation by 1,25-dihydroxyvitamin D3 regulation of protein kinase C levels and activity. AB - The hormone 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] promotes differentiation of a number of cell types including HL-60 promyelocytic leukemia cells. It is now established that protein kinase Cbeta (PKCbeta) plays a critical role in HL-60 cell maturation to a monocyte/macrophage phenotype. In the present study, we investigated the importance of PKCbeta levels and activation in 1,25-(OH)2D3 mediated differentiation of HL-60 cells. Cell differentiation promoted by 1,25 (OH)2D3 at 48 hr was 39 +/- 3% (mean +/- SEM) nitroblue tetrazolium (NBT) positive and at 72 hr it was 35 +/- 2% NBT positive and 70% CD14 positive. Thus, promotion of cell differentiation by 20 nM 1,25-(OH)2D3 treatment was maximal at 48-72 hr. When PKCbeta levels and cell differentiation were assayed at 72 hr, treatment with 20 nM 1,25-(OH)2D3 for the initial 6 hr increased PKCbeta levels by 175% but had little effect on cell differentiation (7 +/- 2% NBT positive; 11% CD14 positive). The effect of ionomycin, a calcium ionophore, on PKCbeta levels and cell differentiation also was examined. Alone, 5 microM ionomycin promoted few cells (3% CD14 positive) to differentiate. In contrast, cells treated with 5 microM ionomycin for 66 hr after a 6-hr pretreatment with 20 nM 1,25-(OH)2D3 resulted in 34 +/- 5% NBT positive cells and 73% CD14 positive cells. Quantitatively, this induction of differentiation was identical to that observed in cultures continuously treated with 1,25-(OH)2D3 (35 +/- 2% NBT positive; 70% CD14 positive). Therefore, ionomycin seemed to replace the requirement for the continuous presence of 1,25-(OH)2D3. Chelerythrine chloride (3 microM), a specific PKC inhibitor, blocked differentiation promoted by 1,25-(OH)2D3 alone (82 +/- 2% inhibition) or in sequence with ionomycin (86 +/- 3% inhibition). Taken together, our data show that the capacity of 1,25-(OH)2D3 to both increase PKCbeta levels and activate PKC is utilized to promote HL-60 cell differentiation. These data further suggest that 1,25-(OH)2D3 has a genomic action to increase PKCbeta levels and also a nongenomic action requiring its continuous presence to promote HL-60 cell differentiation. PMID- 9354592 TI - Prostanoid receptor with a novel pharmacological profile in human erythroleukemia cells. AB - The purpose of this study was to characterize the prostanoid receptors coupled to intracellular calcium in human erythroleukemia (HEL) cells, a cell line with platelet/megakaryocytic characteristics. Both prostaglandin E1 (PGE1) and iloprost increased cyclic AMP (cAMP) in HEL cells, but modulated [Ca2+]i by different mechanisms. Iloprost (10(-9) to 10(-6) M) had no effect on basal [Ca2+]i, but greatly potentiated the increase in [Ca2+]i produced by thrombin. This effect was mimicked by cholera toxin and other Gs-coupled receptors, and involved calcium influx since iloprost had no effect on [Ca2+]i in cells incubated in Ca2+-free buffer. Furthermore, iloprost did not increase the generation of baseline or thrombin-induced inositol phosphates at these concentrations. In contrast, PGE1 (10(-7) to 10(-5) M), but not iloprost, increased basal [Ca2+]i through a pertussis toxin-sensitive mechanism that involved stimulation of inositol phosphate generation and mobilization of intracellular calcium. The order of potencies of other prostaglandins that increased [Ca2+]i was not consistent with known IP, EP, DP, FP, or TP receptors. 11-Deoxy-16,16-dimethyl PGE2 was the most potent of the analogs tested (EC50 = 28 nM). In summary, at least two prostaglandin receptors are functionally coupled to intracellular calcium in HEL cells: a putative IP receptor coupled to Gs proteins that increases cAMP and enhances calcium influx, and a novel prostanoid receptor that evokes calcium mobilization through stimulation of phospholipase C by a pertussis toxin-sensitive pathway. PMID- 9354593 TI - Role of nitroreductases but not cytochromes P450 in the metabolic activation of 1 nitropyrene in the HepG2 human hepatoblastoma cell line. AB - 1-Nitropyrene is an environmental contaminant that is mutagenic in many prokaryotic and eukaryotic systems, including the hypoxanthine-guanosine phosphoribosyl transferase (HGPRT) locus in the human hepatoma cell line HepG2. Metabolism and DNA adduct formation of [3H]1-nitropyrene in the HepG2 were quantified to understand the role of nitroreduction and/or cytochrome P450 mediated C-oxidation of 1-nitropyrene in DNA adduct formation and mutagenicity. In uninduced HepG2 cells, 10 microM [3H]1-nitropyrene was metabolized principally by nitroreduction to 1-aminopyrene (516 pmol/24 hr/10(6) cells), and by cytochrome P450-mediated C-oxidation to K-region trans-dihydrodiols (37 pmol/24 hr/10(6) cells), 1-nitropyren-3-ol (51 pmol/24 hr/10(6) cells), and 1-nitropyren 6-ol and 1-nitropyren-8-ol (77 pmol/24 hr/10(6) cells). Pretreatment of the HepG2 cells for 24 hr with 5 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) resulted in a complete change in the metabolism of [3H]1-nitropyrene, with 1-nitropyren-6-ol and 1-nitropyren-8-ol formation (449 pmol/24 hr/10(6) cells) being 80-fold greater than 1-aminopyrene formation (6 pmol/24 hr/10(6) cells). This increase in C-oxidation of 1-nitropyrene was consistent with increased levels of cytochrome P450 1A. The only DNA adduct detected using the 32P-postlabeling assay in the HepG2 cells administered 1-nitropyrene was N-(2'-deoxyguanosin-8-yl)-1 aminopyrene (dG-C8-AP). Induction of C-oxidative metabolism through TCDD treatment resulted in a concomitant decrease in dG-C8-AP formation. DNA adducts for oxidized 1-nitropyrene metabolites were not detected in the TCDD-treated HepG2 cells administered 1-nitropyrene, which indicates that cytochrome P450 mediated C-oxidative pathways are detoxification pathways in HepG2 cells. PMID- 9354594 TI - Amphotericin B as an intracellular antioxidant: protection against 2,2' azobis(2,4-dimethylvaleronitrile)-induced peroxidation of membrane phospholipids in rat aortic smooth muscle cells. AB - The antifungal activity of amphotericin B (AmB) and its side-effects (e.g. nephrotoxicity and hemolytic action) are suggested to be associated with its prooxidant effects in target cells. To test this hypothesis, we have undertaken studies to examine the role of AmB in oxidative stress in cultured rat aortic smooth muscle cells (SMC) incubated in the absence or in the presence of a lipid soluble azo-initiator of peroxyl radicals, 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN). No changes in the pattern of membrane phospholipids could be detected by two-dimensional high performance thin-layer chromatography (HPTLC) after oxidative stress induced by AMVN in which the cells remained viable, as judged by trypan blue exclusion. To improve the sensitivity of detection of oxidative stress in the cells, cis-parinaric acid (PnA) was incorporated biosynthetically into the membrane phospholipids [using PnA-human serum albumin (hSA) complex]. Incubation of the cells under aerobic conditions in the presence of up to 10 microM AmB showed no significant change in the pattern of PnA-labeled phospholipids, suggesting that AmB was not affecting the oxidative state of the cells. In contrast, treatment with AMVN (0.5 mM, incubation in the dark for 2 hr at 37 degrees--conditions in which the viability of the cells was maintained) caused a significant reduction of all fluorescently labeled phospholipid fractions separated by HPLC. When PnA-labeled cells were subjected to oxidative stress by incubation with 0.5 mM AMVN in the presence of AmB, the loss of fluorescent phospholipids was reduced in a concentration-dependent manner over a concentration range of 0.25 to 10 microM. Thus, AmB does not produce any prooxidant effect but rather acts as an intracellular antioxidant. PMID- 9354595 TI - Noninvolvement of CYP2E1 in the (omega-1)-hydroxylation of fatty acids in rat kidney microsomes. AB - Pyrazole, acetone, and ethanol are known to induce cytochrome P450 2E1 (CYP2E1) and fatty acid (omega-1)-hydroxylation in rat liver microsomes. However, the nature of the P450 enzyme involved in this (omega-1)-hydroxylation has not been clearly established in extrahepatic tissues such as kidney. Four enzymatic activities (hydroxylations of chlorzoxazone, 4-nitrophenol, and two fatty acids) were assayed in kidney microsomal preparations of rats treated with CYP2E1 inducers. Per os treatment resulted in large increases (threefold to fivefold) in the chlorzoxazone and 4-nitrophenol hydroxylations, and up to a ninefold increase when ethanol was administered by inhalation. However, neither the omega hydroxylation nor the (omega-1)-hydroxylation of fatty acids was modified. Immunoinhibition specific to CYP2E1 did not significantly decrease the omega and (omega-1)-lauric acid hydroxylations, while the polyclonal anti-CYP4A1 antibody inhibited in part both the omega- and (omega-1)-hydroxylations. Chemical inhibitions using either CYP2E1 competitive inhibitors (such as chlorzoxazone, DMSO, and ethanol) or P450 mechanism-based inhibitors (such as diethyldithiocarbamate and 17-octadecynoic acid) led to a partial inhibition of the hydroxylations. All these results suggest that fatty acid (omega-1) hydroxylation, a highly specific probe for CYP2E1 in rat and human liver microsomes, is not mediated by CYP2E1 in rat kidney microsomes. In contrast to liver, where two different P450 enzymes are involved in fatty acid omega- and (omega-1)-hydroxylations, the same P450 enzyme, mainly a member of the CYP4A family, was involved in both hydroxylations in rat renal microsomes. PMID- 9354596 TI - A simple procedure for aesthetic correction of the medial epicanthal fold. AB - The author presents a surgical procedure for aesthetic correction of the medial epicanthal fold. The procedure consists of an asymmetrical Z-plasty of the epicanthal fold where a flap is formed from the posterior surface of the epicanthal fold itself. This procedure does not require geometric planning and produces excellent anatomical and functional results, with inconspicuous scars. PMID- 9354597 TI - Extended lid-splitting surgery of marginal eyelid defects. AB - The technique of extended lid-splitting surgery is described. This procedure was used to repair marginal defects of the lower and upper lid, and provided postoperative results that were excellent both functionally and cosmetically, principally because the technique allowed the anatomical structures to be maintained. PMID- 9354599 TI - External ultrasonic treatment of capsular contractures in breast implants. AB - The authors report their experience on the nonsurgical treatment of capsular contractures due to breast implant augmentation mammaplasty. External ultrasonic repeated applications have been applied to 24 patients after closed capsulotomy procedures in order to reduce the recurrency rate. The new ultrasonic device used was based on a 2-MHz generator with a timing adjustable power emission connected to eight transducers designed for breast anatomy. The authors report significant improvement of the closed capsulotomy technique demonstrating a persistent stability of the achieved results in 82% of the treated contractures, even in severe cases (Baker's IV), after a minimum follow-up period of 12 months. Methods of application, technical features of the ultrasonic device, experimental charts, and results obtained on 34 breast implant capsular contractures are reported and discussed. PMID- 9354598 TI - Submental skin: morphohistological study of interest to liposuction. AB - Skin study of the submental region in cadavers made possible the evaluation of its regional characteristics. Sex or race were not reported, but the age group most appropriate to liposuction was considered. In this way, the authors believed that through standards established at the time, results, as to the skin retraction in liposuction, will be able to be analyzed in a comparative form in the future. PMID- 9354600 TI - CAST liposuction: an alternative to brachioplasty. AB - The aging female with excess arm fat and poor skin tone frequently refuses a brachioplasty scar due to permanent detectability. Traditional deep liposuction localized to the posterolateral aspect of the arm frequently leaves sagging, wrinkled skin. Circumferential para-Axillary Superficial Tumescent (CAST) liposuction was developed to maximize skin retraction and create regional harmony by preparatory compartment magnification with dilute lidocaine and epinephrine followed by circumferential treatment of the arm and adjacent areas utilizing superficial and/or subdermal liposuction. Early CAST liposuction results in patients with moderate fat and excess skin revealed excellent skin retraction. CAST liposuction was then offered as the first of two stages to patients with excess fat and poor skin tone to avoid or shorten the brachioplasty scar. Twenty six patients underwent CAST liposuction with 9-22 months follow up. Only two patients (7.7%) eventually required brachioplasty. Although postoperative seromas were frequent (38.5%) and preexisting skin wrinkling usually returned, the final result is acceptable to the vast majority of patients (84.6%) who refuse a brachioplasty scar. PMID- 9354601 TI - Vertical mammaplasty as secondary surgery after other techniques. AB - Vertical mammaplasty, a technique that avoids submammary scars, has proved to be a reliable method of breast reduction because it is adaptable to most cases and produces beautiful and durable results. What about secondary cases? In the last 14 cases referred for secondary mammaplasty, at 1-19 years after their initial surgery, patients' indications were poor shape (14), visible and improperly located scars (9), excess volume (8), asymmetry of the areolas (5) or the breasts (1), insufficient volume (2), and asymmetry with reconstructed breast (2). The original scars were inverted T (10), periareolar (2), oblique (1) or vertical (1). Their appearance was a concern for nine patients. All patients but one, who had long submammary scars surrounded by heavy stitch marks requiring correction, could benefit from a vertical mammaplasty. This avoided long months of scar redness and visibility along the submammary folds. Good symmetry and shape could be obtained in all cases by adjusting the markings to the needs. Liposuction was a great help to remove volume without endangering the blood supply of the areolas, possibly transforming reductions in simple mastopexies. PMID- 9354602 TI - Nummular nipple hypertrophy and repair as part of an aesthetic nipple-areola unit. AB - The author suggests that an aesthetically pleasing ratio between nipple and areola diameter exists which should always be taken in consideration during nipple and areola reconstruction. In a study of 40 nipple-areola complexes of 20 healthy, nulliparous, Caucasian female volunteers with a mean age of 25.5 years, the average nipple diameter measured 28% of the areola diameter, that is, a ratio of 1:3.6. A hitherto undescribed form of macrothelia is presented in which the nipple width rather than the projection (length) is increased. A successful technique for reconstruction is described, based on the new method of assessing the aesthetic relations within the nipple-areola complex and known anatomy. PMID- 9354603 TI - Breast shape malformations. AB - Breast shape malformations are deformities of the breast contour involving the inferior breast pole. They range from minor forms such as a high submammary fold to major forms such as the tubular breast. The malformation implies an altered relationship between the mammary parenchyma and the superficial fascial system (SFS). The surgical correction consists of the releasing of the gland from all the connections with the surrounding tissues. In some cases, once the glandular tissue is freed from the constricting SFS, the parenchyma spreads revealing a sufficient volume to obtain a normal breast. If hypoplasia of the breast or inferior quadrant hypoplasia is present, the use of an implant is mandatory. PMID- 9354604 TI - Augmentation mammoplasty: personal evolution of the concepts looking for an ideal technique. AB - The purpose of this paper is to report our personal experience in the field of augmentation mammoplasty. This experience is based on over 15 years in practice and working with more than 400 cases using different types of prostheses (single lumen gel-filled, single-lumen saline-filled, double-lumen, smooth or texturized surfaces), different routes (submammary, periareolar, transaxillary), and different locations of the implant (complete submuscular, subglandular, subpectoral). Our present preference is for a partial submuscular (subpectoral) augmentation mammoplasty through an inferior periareolar route. The results of 91 consecutive patients operated on with this technique from January, 1990 to December, 1994, during the blow-up of the controversy on silicone, are reported. PMID- 9354605 TI - Sixth-month morphoauricular syndrome. AB - Based upon a study of the embryologic anatomy of the external ear, the authors have suggested that the antitragus muscle be called the auricularis inferioris muscle which has a double nerve supply and a double function. Abnormal traction of this muscle can vary the final shape of the ear, dragging the border of the antitragus downward and everting it. This scaphoid and prominent ear deformity can be corrected by precise surgery described here by the authors. PMID- 9354606 TI - Repair of the partial loss of the helix. AB - This article presents a new technique for the repair of small and medium-sized defects of the helix using a retroauricular flap whose advance is obtained by the displacement of the auricular fold. PMID- 9354607 TI - Restoration of the lip roll with Gore-Tex. AB - Gore-Tex has been used by the author for the past 2 years to enhance the vermillion border of the lips by emphasizing that a natural pocket exists between the skin and the orbicularis muscle all along the white roll where a Gore-Tex implant can be easily placed. The overlying skin is thick providing a good protection of this material, and such a thickness necessitates that this protection is provided. If a complication occurs from its use, then the implant can be easily removed without any damage to the adjacent tissues. PMID- 9354608 TI - Evaluation of lip augmentation with Gore-Tex facial implant. AB - Although numerous materials, including autogenous, homogeneous, and alloplastic materials, have been used for lip augmentation with varying degrees of success, no ideal one has been found to achieve a soft and long-lasting result. Gore-Tex implant has been successfully used in cardiovascular surgery. So far, it has not been used much in lip augmentation. In this study, a Gore-Tex SAM facial implant (1.8 mm in diameter) was used for lip augmentation in 23 lips of 17 female patients. In 10 consecutive lips with three segments of the implant each, a computer-assisted imaging system was used to scrutinize the results. The results have shown that the three segments of the implant enhanced the lip projection with about 0.98 mm in mean (p < 0.01) and the exposed vermillion width with about 1. 94 mm (p < 0.01) in the over 6 months follow-ups. No major complications, only some minor ones, were seen. For lip augmentation, we find that only minor changes can be achieved with three segments of the Gore-Tex facial implant. We feel that it is safe and believe it gives a permanent result which is not the case with collagen injection and usually not the case with lipofilling despite reinjections. Furthermore, we consider the implant quite expensive at present. PMID- 9354609 TI - Successful surgery of multiple recurrent basal cell carcinomas guided by photodynamic diagnosis. AB - We describe a 56-year-old Caucasian man with history of multiple regressed basal cell carcinomas. During the last 20 years approximately 200 histologically proven basal cell carcinomas preferentially localized on the face were surgically treated. Several large skin grafts were necessary to cover the extensive tissue defects on the face and scalp. Although all excised tissues were histologically proven to be basal cell carcinomas with tumor-free margins, new tumors developed in proximity to the skin graft margins. The dissemination of the new tumors made it difficult to perform additional invasive operation procedures without influencing the cosmetic result. Thus, we used photodynamic diagnosis to improve detection and demarcation of the neoplastic tissues. This procedure facilitated surgical planning and enabled primary in toto excisions. Surgical trauma and a number of interventions were thus minimized with the consequence of improved cosmetic and functional results. PMID- 9354611 TI - Long-term follow-up in treatment of solar lentigo and cafe-au-lait macules with Q switched ruby laser. AB - A Q-switched ruby laser was used for treatment of 10 patients with solar lentigo and 12 patients with cafe-au-lait macules. In this study, the lesions were treated with the laser at a rate of 6 J/cm2. The patients were observed for 10-21 months with an average of 13.8 months after the final session. Solar lentigos were treated once or twice, and the response rate was 70%. Cafe-au-lait macules were treated one to six times, and the response rate was 33%. Side effects, such as hyperpigmentation and scar formation, were rarely seen. Therefore, Q-switched ruby laser treatment is an effective treatment for epidermal pigmented lesions; however, in patients with cafe-au-lait macules, the responses to the treatment varied, and a repigmentation was seen in 50% of these patients. Thus, long-term follow-up is required for patients with cafe-au-lait macules. PMID- 9354610 TI - Aesthetic reconstruction of burn alopecia by using expanded hair-bearing scalp flaps. AB - Tissue expansion is one of the most important armamentaria for aesthetic scalp reconstruction after burn; however, the proper way to employ this technique for the scalp reconstruction usually presents a challenge to the plastic surgeon, especially in the case of a "sideburn" scenario or a large lesion, as with, for example, hemiscalp alopecia. In this article, 11 patients, with different degrees of hair-bearing scalp loss as a result of burn, and including four patients with hemiscalp alopecia were successfully treated by using tissue expansion. The results show that tissue expansion is a simple, safe, and efficient technique for aesthetic scalp reconstruction. Versatile design of the expanded scalp flap can distribute the expanded hair-bearing scalp properly in the reconstructed recipient site. PMID- 9354612 TI - Actin-tropomyosin activation of myosin subfragment 1 ATPase and thin filament cooperativity. The role of tropomyosin flexibility and end-to-end interactions. AB - Tropomyosin (Tm) bound to actin induces cooperative activation of actomyosin subfragment 1 (actin-S1) ATPase, observed as a sigmoid ATPase vs [S1] dependence. The activation is much steeper for gizzard muscle Tm (GTm) than for rabbit skeletal Tm (RSTm). To investigate if this greater cooperativity is due to increased communication between GTms along the thin filament, we studied effects of S1 binding on the state of actin-Tm using the fluorescence of pyrene-labeled Tm. Kinetic and equilibrium studies provided values for n, the apparent cooperative unit size [Geeves, M. A., and Lehrer, S. S. (1994) Biophys. J. 67, 273]. We report comparative studies of Tm-actin-S1 ATPase with values of n using GTm, RSTm, and 5aTm, a 1/7 shorter nonmuscle Tm from rat fibroblast cells [Pittenger, M. F., et al. (1994) Curr. Opin. Cell Biol., 6, 96]. 5aTm and GTm produce similar cooperative activation of actin-S1 ATPase and have similar n values that are 2-fold greater than RSTm, indicating a correlation between ATPase activation and n value. This appears to be due to the similarity of the C terminal amino acid sequences of 5a and GTm which produce strong end-to-end interactions. The results are discussed in terms of a continuous flexible Tm strand on the actin filament. PMID- 9354613 TI - Melittin-induced inhibition and aggregation of Ca-ATPase in skeletal muscle sarcoplasmic reticulum: a comparative study. AB - Incubation of melittin with sarcoplasmic reticulum membranes at pH 7. 0 and different melittin:Ca-ATPase molar ratios results in the progressive loss of enzyme activity. At high melittin:Ca-ATPase molar ratios (10:1 and 30:1), enzyme inhibition may be described by a biexponential curve. At pH 7.0, the values of the pseudo-first-order rate constants are 1.0 and 0.1 min-1 for the fast and slow phases of inhibition, respectively, at a melittin:Ca-ATPase molar ratio of 30:1. At pH 6.0 and a melittin:Ca-ATPase molar ratio of 30:1, melittin does not inhibit Ca-ATPase. Melittin-induced aggregation of Ca-ATPase molecules was studied using cupric phenanthroline as a chemical cross-linking agent. At a melittin:Ca-ATPase molar ratio of 5:1, aggregation of Ca-ATPase protein was not observed; however, the loss of enzyme activity was about 30% after 30 min. At melittin:Ca-ATPase molar ratios of 10:1 and 30:1, significant aggregation of Ca-ATPase protein takes place. The rate of Ca-ATPase aggregation is much lower than the rate of enzyme inhibition. At melittin:Ca-ATPase molar ratios of 10:1 and 30:1, the rate of Ca ATPase protein aggregation is close to that for the slow phase of enzyme inhibition. At pH 6.0 and a melittin:Ca-ATPase molar ratio of 30:1, significant aggregation of Ca-ATPase occurs. It is concluded that melittin induces both Ca ATPase inhibition and aggregation. These two processes may occur simultaneously, but under some conditions either inhibition or aggregation takes place independently of each other. Therefore, the aggregation of Ca-ATPase induced by melittin is not necessary for enzyme inhibition. PMID- 9354614 TI - Purification and characterization of an alpha 1 beta 2 isoform of CapZ from human erythrocytes: cytosolic location and inability to bind to Mg2+ ghosts suggest that erythrocyte actin filaments are capped by adducin. AB - CapZ ("capping protein") is a heterodimeric actin capping protein that blocks actin filament assembly and disassembly at the fast growing (barbed) filament ends and is proposed to function in regulating actin filament dynamics as well as in stabilizing actin filament lengths in muscle and nonmuscle cells. We show here that erythrocytes contain a nonmuscle isoform of capZ (EcapZ) that is present exclusively in the cytosol and is not associated with the short actin filaments in the erythrocyte membrane skeleton. This is unlike other cell types where capZ is associated with cytoskeletal actin filaments and suggests that cytosolic EcapZ may be inactive, or alternatively, that the barbed ends are capped by adducin, a membrane skeleton protein that was shown recently to cap actin filament barbed ends in vitro [Kuhlman, P. A., Hughes, C. A., Bennett, V., & Fowler, V. M. (1996) J. Biol. Chem. 271, 7986]. To distinguish between these possibilities, we purified EcapZ from erythrocyte cytosol and characterized its biochemical and functional properties. Two-dimensional gel electrophoresis and western blotting reveals the EcapZ subunit composition to be alpha1beta2, as described for capZ from many other nonmuscle cells, with no evidence for posttranslational modifications. Purified EcapZ is fully functional in blocking actin elongation from barbed filament ends (Kcap approximately 1-5 nM) as well as in nucleating actin polymerization. Furthermore, cytosolic EcapZ binds to actin filament barbed ends, indicating that sequestering of EcapZ by a cytosolic inhibitory factor or insufficient amounts of EcapZ in cytosol also cannot account for its absence from the membrane skeleton. To test directly whether the barbed ends of the erythrocyte actin filaments were already capped, we measured binding of purified EcapZ to isolated membranes. Purified EcapZ does not cosediment with membranes prepared by hypotonic lysis in the presence of magnesium, suggesting that the barbed ends of the erythrocyte actin filaments are capped under these conditions but not by EcapZ. In contrast, purified EcapZ stoichiometrically reassociates with all the actin filament barbed ends in membranes prepared by hypotonic lysis in 5 mM sodium phosphate, pH 8.0 (5P8), conditions in which the barbed filament ends were previously reported to be uncapped. Comparison of the amounts of adducin associated with membranes prepared in the presence and absence of magnesium reveals that 60-80% of the adducin dissociates from the membrane during hemolysis and washing in 5P8 buffer, suggesting that the barbed ends become artifactually uncapped due to loss of adducin. The erythrocyte actin filaments may thus represent a specialized class of membrane-associated actin filaments that are capped by adducin instead of capZ. PMID- 9354615 TI - Purification, characterization, and synthesis of three novel toxins from the Chinese scorpion Buthus martensi, which act on K+ channels. AB - Three novel toxins belonging to the scorpion K+ channel-inhibitor family were purified to homogeneity from the venom of the Chinese scorpion Buthus martensi. They have been identified according to their molecular mass (3800-4300 Da) and their neurotoxicity in mice and characterized as 37-amino acid peptides. One of them shows 81-87% sequence identity with members of the kaliotoxin group (named BmKTX), whereas the other two, named BmTX1 and BmTX2, show 65-70% identity with toxins of the charybdotoxin group. Their chemical synthesis by the Fmoc methodology allowed us to show that BmKTX, unlike BmTX1 and BmTX2, possesses an amidated C-terminal extremity. Toxicity assays in vivo established that they are lethal neurotoxic agents in mice (LD50s of 40-95 ng per mouse). Those toxins proved to be potent inhibitors of the voltage-gated K+ channels, as they were able to compete with [125I]kaliotoxin for its binding to rat brain synaptosomes (IC50s of 0.05-1 nM) and to block the cloned voltage-gated K+ channel Kv1.3 from rat brain, expressed in Xenopus oocytes (IC50s of 0.6-1.6 nM). BmTX1 and BmTX2 were also shown to compete with [125I]charybdotoxin for its binding to the high conductance Ca2+-activated K+ channels present on bovine aorta sarcolemmal membranes (IC50s of 0.3-0.6 nM). These new sequences show multipoint mutations when compared to the other related scorpion K+ channel toxins and should prove to be useful probes for studying the diverse family of K+ channels. PMID- 9354616 TI - Catalytic domain structure of vampire bat plasminogen activator: a molecular paradigm for proteolysis without activation cleavage. AB - The saliva of the blood-eating vampire bat Desmodus rotundus contains plasminogen activators (PAs) that maintain the fluidity of the prey's blood by activating plasminogen and dissolving developing fibrin clots. D. rotundus salivary PAs (DSPAs) are composed of evolutionarily conserved domains reminiscent of human tissue-type PA (tPA), but their catalytic domain lacks a plasmin-sensitive "activation cleavage site". Despite this, all DSPAs are intrinsically active and enormously stimulated in the presence of fibrin. The recombinant catalytic domain of DSPAalpha1 has been crystallized in a covalent complex with Glu-Gly-Arg chloromethyl ketone and its structure solved at 2.9 A resolution. The structure is similar to that of activated two-chain human tPA. Despite its single-chain status, the activation domain is observed in an enzymatically active conformation, with a functional substrate binding site and active site accommodating the peptidylmethylene inhibitor. The activation pocket, which normally receives the N-terminal Ile16, is occupied by the side chain of Lys156, whose distal ammonium group makes an internal salt bridge with the carboxylate group of Asp194. Lys156 is in a groove shielded from the bulk solvent by the intact "activation loop" (Gln10-Phe21), favoring Lys156-Asp194 salt bridge formation and stabilization of a functional substrate binding site. Together with the characteristic 186 insertion loop, the activation loop could act as a switch, effecting full single-chain enzymatic activity upon binding to fibrin. PMID- 9354617 TI - Nonpolar interactions of thrombin S' subsites with its bivalent inhibitor: methyl scan of the inhibitor linker. AB - We have designed bivalent thrombin inhibitors, consisting of a nonsubstrate type active site blocking segment, a hirudin-based fibrinogen recognition exosite blocking segment, and a linker connecting these segments. The inhibition provided by the bivalent inhibitors with various linker lengths revealed that a minimum of 15 atoms was required for simultaneous binding of the two blocking segments of the inhibitor to thrombin without significant distortion. The crystal structure of the inhibitors with a 16-atom linker showed some conformational flexibility in the linker portion which still lies deep in the groove joining the active site and the fibrinogen recognition exosite. Since the thrombin S' subsites are not well characterized, we designed a new strategy to search for possible nonpolar interactions between the linker and the thrombin S' subsites. This strategy, the "methyl scan", is based on the incorporation of a methyl side chain at each atom position of the linker by using sarcosine, D,L-alanine, D,L-3-aminoisobutyric acid, or N-methyl-beta-alanine. The methyl groups on the second and the eighth atom positions of the linker, which correspond to the side chains of the P1' and the P3' residues, respectively, improved the affinity of the inhibitors significantly. Further study of the stereospecificity showed that L-Ala at the P1' residue and D-Ala at the P3' residue preferably improved the affinity of the inhibitors 20- and 25-fold, respectively. Molecular modeling calculations using a methyl probe were also carried out to identify favorable nonpolar interacting sites on the thrombin surface. Two sites were identified in the vicinity of the P1' and the P3' residues, supporting the validity of the methyl scan method. Thus, this study has improved our understanding of the interactions taking place in this groove. In particular, we have been able to show that some specific structural features, such as hydrophobic complementarity between the linker and the thrombin S' subsites, could be exploited and make these inhibitors trivalent. PMID- 9354618 TI - Tertiary and quaternary structures of 0.19 alpha-amylase inhibitor from wheat kernel determined by X-ray analysis at 2.06 A resolution. AB - The crystal structure of 0.19 alpha-amylase inhibitor (0.19 AI) from wheat kernel was determined by the multiple-isomorphous replacement method coupled with density modification and noncrystallographic symmetry averaging and then refined by simulated annealing using diffraction data to 2.06 A resolution (R = 18.7%, free R = 22.3%). The asymmetric unit has four molecules of 0.19 AI, each comprised of 124 amino acid residues. Electron density for residues 1-4 and 69-77 is absent in all subunits, probably because of the intrinsic flexibility of these segments. Each subunit has four major alpha-helices and one one-turn helix which are arranged in the up-and-down manner, maintaining the favorable packing modes of the alpha-helices. 0.19 AI, however, has two short antiparallel beta-strands. All 10 cysteine residues in 0.19 AI form disulfide bonds (C6-C52, C20-C41, C28 C83, C42-C99, and C54-C115), consistent with the assignments made biochemically for 0.28 AI from wheat kernel and by NMR analysis of the bifunctional alpha amylase/trypsin inhibitor from ragi seeds (RBI). The disulfide bond patterns in these AIs are similar to those in the hydrophobic protein from soybean (HPS), which lack only the bond corresponding to C28-C83 in 0.19 AI. Extensive interactions occurred between particular pairs of 0.19 AI subunits, mainly involving hydrophobic residues. Comparisons of the structures of 0.19 AI, RBI, and HPS showed that the arrangements of the major alpha-helices are similar but the conformations of the remaining residues differ markedly. The present X-ray analysis for 0.19 AI and the NMR analysis for RBI suggest that all the AIs in this family have a common fold. The alpha-amylase binding site is discussed on the basis of the tertiary and quaternary structures of 0.19 AI together with biochemical and spectroscopic data for AIs. PMID- 9354619 TI - Efficient characterization of collective motions and interresidue correlations in proteins by low-resolution simulations. AB - A low-resolution model is used together with recently developed knowledge-based potentials for exploring the dynamics of proteins. Configurations are generated using a Monte Carlo/Metropolis scheme combined with a singular value decomposition technique (SVD). The approach is shown to characterize the cooperative motions in good detail, at least 1 order of magnitude faster than atomic simulations. Trajectories are partitioned into modes, and the slowest ones are analyzed to elucidate the dominant mechanism of collective motions. Calculations performed for bacteriophage T4 lysozyme, a two-domain enzyme, demonstrate that the structural elements within each domain are subject to strongly coupled motions, whereas the motions of the two domains with respect to each other are strongly anticorrelated. This type of motion, evidenced by the synchronous fluctuations of the domain centroids by up to +/-4.0 A in opposite directions, is comparable to the movements observed by recent spin-labeling experiments in solution. The potential of mean force governing these fluctuations is shown to be anharmonic. The beta-sheet region at the N-terminal domain and the helix E in the C-terminal domain are identified as regions important for mediating cooperative motions and, in particular, for the opening and closing of the active-site cleft between the domains. Residues Leu66-Phe67 in the central helix C stop the propagation of correlated motions between the domains. There is a correlation between the groups involved in highly cooperative motions revealed by simulations and the highly protected regions during unfolding measured by pulsed H/D exchange and 2-D NMR. PMID- 9354620 TI - Nucleotide cofactor-dependent structural change of Xenopus laevis Rad51 protein filament detected by small-angle neutron scattering measurements in solution. AB - Rad51 protein, a eukaryotic homologue of RecA protein, forms a filamentous complex with DNA and catalyzes homologous recombination. We have analyzed the structure of Xenopus Rad51 protein (XRad51.1) in solution by small-angle neutron scattering (SANS). The measurements showed that XRad51.1 forms a helical filament independently of DNA. The sizes of the cross-sectional and helical pitch of the filament could be determined, respectively, from a Guinier plot and the position of the subsidiary maximum of SANS data. We observed that the helical structure is modified by nucleotide binding as in the case of RecA. Upon ATP binding under high-salt conditions (600 mM NaCl), the helical pitch of XRad51.1 filament was increased from 8 to 10 nm and the cross-sectional diameter decreased from 7 to 6 nm. The pitch sizes of XRad51.1 are similar to, though slightly larger than, those of RecA filament under corresponding conditions. A similar helical pitch size was observed by electron microscopy for budding yeast Rad51 [Ogawa, T., et al. (1993) Science 259, 1896-1899]. In contrast to the RecA filament, the structure of XRad51.1 filament with ADP is not significantly different from that with ATP. Thus, the hydrolysis of ATP to ADP does not modify the helical filament of XRad51.1. Together with our recent observation that ADP does not weaken the XRad51.1/DNA interaction, the effect of ATP hydrolysis on XRad51.1 nucleofilament should be very different from that on RecA. PMID- 9354622 TI - Effect of additional unpaired bases on the stability of three-way DNA junctions studied by fluorescence techniques. AB - Fluorescence melting experiments were carried out to determine the relative stability of three-way DNA junctions with and without extrahelical adenine nucleotides in one strand at the branch point of the junction (i.e., An bulges where n = 0, 1, 2, and 3). The oligonucleotides were labeled with chromophores at the 5' ends of the strands. The progress of the thermal denaturation was followed by monitoring the fluorescence intensities and anisotropies of the dyes and the fluorescence resonance energy transfer between the two dyes. The results of the thermal denaturation experiments are interpreted and discussed in terms of either two-state thermodynamic models or statistical models for the thermal denaturation. The junctions all melt at the same temperature (at equal concentrations) within the error of the Tm determination, regardless of the presence, or absence, of the bulge. It is suggested that the denaturation of the helical arms begins primarily at the free ends of the helical arms and proceeds toward the branch point. The junctions, all which have 10 base pairs in each arm, possess thermal denaturation characteristics similar to duplexes with 20 arms. This leads to the proposition that for these junctions an important molecular parameter that controls the stability of the junctions is the number of base pairs between neighboring arms. The melting profiles obtained by monitoring the tetramethylrhodamine fluorescence are found to depend strongly on the nucleotide sequence in the single-stranded region. PMID- 9354621 TI - Global structure of three-way DNA junctions with and without additional unpaired bases: a fluorescence resonance energy transfer analysis. AB - The structure of three-way DNA junctions with and without extrahelical adenine nucleotides in one strand at the branch point of the junction (i.e., An bulges with n = 0, 1, 2, and 3) has been investigated by fluorescence resonance energy transfer. The structure of the junction without bulged nucleotides was found to have a symmetric trigonal geometry. With bulges, the arrangement of the arms becomes asymmetrical. The energy transfer results suggest a model of bulged junctions where the angle between two of the arms is significantly smaller than between the other two pairs of arms. The acute angle becomes smaller as the number of nucleotides in the bulge increases. The FRET efficiencies of the junctions are the same in the presence of Mg++ and Na+ ions. PMID- 9354623 TI - The association of complementary ribonucleic acids can be strongly increased without lowering Arrhenius activation energies or significantly altering structures. AB - The association rates of complementary nucleic acids can be increased by 2-3 orders of magnitude in vitro by cellular proteins and low molecular weight compounds including cetyltrimethylammonium bromide (CTAB). In this work, we provide experimental evidence that the CTAB-mediated enhancement of RNA-RNA annealing by approximately 3 orders of magnitude is due to a favorable activation entropy (DeltaS) and not due to a decrease of the Arrhenius activation energy (Ea) nor to major structural changes of the RNA. Two alternative models for the CTAB-facilitated RNA-RNA annealing will be discussed. First, CTAB could form a positively charged liquid matrix which could steer complementary RNA molecules and thereby increase their collision frequency and annealing rate. Second, increased annealing rates could be explained by stabilization of a non-base specific precomplex of both complementary RNA molecules in solution. PMID- 9354624 TI - Transient-state kinetics of the reaction of aspartate aminotransferase with aspartate at low pH reveals dual routes in the enzyme-substrate association process. AB - In aspartate aminotransferase, the coenzyme pyridoxal 5'-phosphate forms a Schiff base with the epsilon-amino group of Lys258. The pH dependency of the steady state kinetics of the overall reaction had indirectly suggested that the Schiff base-unprotonated form of the enzyme (EL) is the active species that binds the monoanionic form of aspartate (SH+), the predominant species of the substrate in solution. In order to obtain direct information on the association process, we carried out transient-phase kinetics of the first half-reaction of the enzyme with aspartate at various pH. The disappearance of EL (lambdamax = 358 nm) was fast and independent of pH, but the disappearance of ELH+ (Schiff-base-protonated form, lambdamax = 430 nm) was slow and dependent on pH. At pH values below 6.8 and low concentrations of aspartate, the results could be interpreted to indicate that EL reacts rapidly with SH+ to form the pyridoxamine 5'-phosphate form of the enzyme (EM), and the reaction of ELH+ proceeds via the route ELH+ right arrow over left arrow EL right arrow over left arrow EM, where the first step was found to be rate limiting from the pH jump/drop study of the enzyme. At higher pH values, the rate of disappearance of ELH+ became larger than expected from the above scheme. This deviation became apparent with increasing pH, and could be excellently explained if we consider that it is due to the reaction of ELH+ with the dianionic form of aspartate (S). Thus, the formation of the Michaelis complex of aspartate aminotransferase and aspartate can proceed via two routes; route A is the association of EL with SH+ to form EL.SH+, which converts intramolecularly to ELH+.S, and route B is the association of ELH+ with S to form ELH+.S directly. ELH+.S is the prerequisite structure for further processing of the substrate by the enzyme. The reactions of EM and oxo acids yielded almost exclusively EL and SH+, and therefore route B does not seem to play an essential role in the overall reactions of the enzyme. Route B, however, may be important in the reaction mechanisms of other pyridoxal 5'-phosphate enzymes which have only the ELH+ form. PMID- 9354625 TI - Glycosaminoglycans mediate cell surface oligomerization of chemokines. AB - Chemokines are 8-10 kDa proteins involved in the control of leukocyte trafficking and activation. In free solution, chemokines are monomers at physiologic concentrations, although many multimerize at higher concentrations. Cell surface heparan sulfate may sequester chemokines, increasing their local concentrations and facilitating their binding to receptors expressed on leukocytes. In competitive binding assays using immobilized heparin, a 2-3-fold increase in the bound radiolabeled chemokine was seen with increasing concentrations of unlabeled chemokine in the nanomolar range. Unlabeled chemokine concentrations between 0.25 and 50 microM were needed to compete the bound radioactivity. This biphasic competition curve was not seen for N-methyl-L25 IL-8, a variant of IL-8 which is unable to dimerize. In addition, complexes of chemokine and heparin eluted from gel filtration columns with apparent molecular masses of 33-60 kDa, suggesting that chemokine multimerization had occurred. The physiological relevance of this multimerization process was seen from studies using human endothelial cells. The endothelial cell binding sites for IL-8, RANTES, and MCP-1 were deduced to be glycosaminoglycans since competition assays showed the biphasic curves and micromolar IC50 values seen in studies with immobilized heparin, and mRNA for known chemokine receptors was not detected. Furthermore, digestion of endothelial cell monolayers with glycosaminidases decreased chemokine binding by up to 80%. Glycosaminoglycans can act as modulators of the ligand binding affinity of chemokine receptor-bearing cells. Removal of glycosaminoglycans from CHO cells expressing chemokine receptors CXCR1, CCR1, or CCR2 resulted in 40-70% decreases in the binding of RANTES, MCP-1, IL-8, and MIP-1alpha. Our data show that cell surface glycosaminoglycans induce polymerization of chemokines, increasing their local concentration and therefore enhancing their effects on high-affinity receptors within the local microenvironment. PMID- 9354626 TI - Determinants of calcineurin binding to model membranes. AB - The biochemical factors that lead to membrane targeting of the Ser/Thr protein phosphatase calcineurin were examined using model phospholipid membranes. The interaction of myristoyl- and non-myristoylcalcineurin with lipid surfaces was investigated as a function of negatively charged phospholipids, diacylglycerol, Ca2+, and calmodulin. The data indicate that calcineurin binding to phospholipid monolayers both is myristoyl-independent and is mediated by anionic phospholipids and/or diacylglycerol. Although the effect of Ca2+ on calcineurin-lipid binding is minor, calmodulin altered the binding of calcineurin to the lipid membrane in a Ca2+-dependent manner. Experiments with a constitutively active form of calcineurin that does not bind calmodulin indicated that the effect required the interaction of calcineurin with calmodulin. Our results suggest that phosphatidylserine, diaclyglycerol, and calmodulin may mediate the lipid binding properties of calcineurin in vivo. PMID- 9354628 TI - Monitoring structural stability of trypsin inhibitor at the submolecular level by amide-proton exchange using Fourier transform infrared spectroscopy: a test case for more general application. AB - Combining the information on the secondary structure content as present in the shape of a protein amide I infrared band with the approach of monitoring amide proton exchange using infrared spectroscopy, we have been able to investigate the structural stability of different components present in a protein, which are shown to be correlated to the different classes of secondary structures. For this purpose, the changes in intensity in different regions of the amide I have been detected upon exposure of the protein to a 2H2O environment, revealing four separate classes of exchanging components. As a test case for the approach described in this work, the amide-proton exchange of hydrated protein films of bovine pancreatic trypsin inhibitor has been studied using infrared spectroscopy, and is compared to literature data obtained by other techniques. A slow amide proton exchange is observed for a class correlated to the beta-strands present in the protein, with protection of amide-protons for more than 19 h. Another class, which has been assigned to mainly helical residues, shows much less protection from exchange. The distribution function of the exchange rates of a class linked to the beta-turns displays five times faster exchange rates compared to those found for the majority of the helical residues, but they are still ten times slower compared to a class which we defined to represent the nonstructured parts of the protein. PMID- 9354627 TI - Catalytic role of monovalent cations in the mechanism of proton transfer which gates an interprotein electron transfer reaction. AB - Within the methylamine dehydrogenase (MADH)-amicyanin protein complex, long range intermolecular electron transfer (ET) occurs between tryptophan tryptophylquinone (TTQ) of MADH and the type I copper of amicyanin. The reoxidations of two chemically distinct reduced forms of TTQ were studied, a quinol (O-quinol) generated by reduction by dithionite and the physiologically relevant aminoquinol (N-quinol) generated by reduction by methylamine. The latter contains a substrate derived amino group which displaces the C6 carbonyl oxygen on TTQ. ET from N quinol MADH to amicyanin is gated by the transfer of a solvent exchangeable proton [Bishop, G. R., & Davidson, V. L. (1995) Biochemistry 34, 12082-12086]. The factors which influence this proton transfer (PT) reaction have been examined. The rate of PT increases with increasing pH and with increasing salt concentration. The salt effect is due to specific monovalent cations and is not a general ionic strength effect. The rate enhancements by pH and cations do not reflect an elimination of the PT step that gates ET. Over the range of pH from 5.5 to 9.0 and with cation concentrations from 0 to 200 mM, the observed rate of the redox reaction is still that of PT. This is proven by kinetic solvent isotope effect studies which show that a primary isotope effect persists even at the highest values of pH and cation concentration. A model is presented to explain how specific cations contribute to catalysis and influence the rate of PT in this reaction. The pH dependence is attributed to an ionizable group that is involved in cation binding. The effect of the cation is stabilization of a negatively charged reaction intermediate that is formed during the deprotonation of the N quinol, and from which rapid ET to the copper of amicyanin occurs. The relevance of these findings to other enzymes which exhibit reaction rates that are influenced by monovalent cations is also discussed. PMID- 9354630 TI - Pulse radiolysis studies on cytochrome cd1 nitrite reductase from Thiosphaera pantotropha: evidence for a fast intramolecular electron transfer from c-heme to d1-heme. AB - Electron transfer within cytochrome cd1 from Thiosphaera pantotropha was investigated by the technique of pulse radiolysis. The reduction of the heme centers in this nitrite reductase occurred in two phases as judged from kinetic difference spectra. In the faster phase, radiolytically generated N methylnicotinamide (NMA) radicals selectively reduced the c-heme of the enzyme. From the absorbance increase at 420 nm, a characteristic of formation of the ferrousc-heme, the second-order rate constant for this electron transfer process was estimated to be 3.8 x 10(9) M-1 s-1 at pH 7.0. In the slower phase, a decrease of absorption around 420 and 550 nm, corresponding to a reoxidation of the c-heme, was accompanied by an increase of absorption around 460 and 640 nm, characteristic of formation of the reduced d1-heme. This indicated that an intramolecular electron transfer from the c-heme to the d1-heme occurred. The first-order rate constant of this process was calculated to be 1.4 x 10(3) s-1 at pH 7.0 and was independent of the enzyme concentration. In the presence of nitrite the interheme electron transfer rate was not affected, but on a time scale of seconds a new species associated with the d1-heme, having an absorption maximum at 640 nm, was detected and is proposed to reflect ligand binding to this heme. These results suggest the role of the c-heme as the electron acceptor site in cytochrome cd1 and in mediating the electron transfer to the catalytic site of the enzyme. Moreover, the fast interheme electron transfer rate argues against this process being the rate determining step in catalysis. PMID- 9354629 TI - Amide-proton exchange of water-soluble proteins of different structural classes studied at the submolecular level by infrared spectroscopy. AB - For eleven films of various water-soluble alpha-, beta-, alpha-/beta-, and alpha +beta-proteins, the amide-proton exchange, initiated by exposure of the protein film to 2H2O, has been monitored using infrared spectroscopy. The approach to obtain the kinetics of exchange for four different classes of amide protons, correlating to the different secondary structure types, has been described in detail in the preceding paper. In this work the more general applicability of the approach is illustrated by testing it for different types of proteins. The results obtained are shown not only to be comparable to reported time-resolved nuclear magnetic resonance data (as in the case of myoglobin, phospholipase A2, lysozyme, and cytochrome c), or to the more qualitative data obtained by neutron diffraction (trypsin, ribonuclease S, papain, and subtilisin BPN'), but the infrared approach us also provides with quantitative detailed insight on the distribution of exchange rate constants at the submolecular level of proteins, too complex to be studied by other techniques, as for tetrameric hemoglobin, and of proteins in which exchange is too fast to be detected by these other techniques, as is shown in this work for alpha-casein and apocytochrome c. PMID- 9354631 TI - Evaluation of the role of specific acidic amino acid residues in electron transfer between the flavodoxin and cytochrome c3 from Desulfovibrio vulgaris. AB - A hypothetical model for electron transfer complex between cytochrome c3 and the flavodoxin from the sulfate-reducing bacteria Desulfovibrio vulgaris has been proposed, based on electrostatic potential field calculations and NMR data [Stewart, D. E., LeGall, J. , Moura, I., Moura, J. J. G., Peck, H. D., Jr., Xavier, A. V., Weiner, P. K., & Wampler, J. E. (1988) Biochemistry 27, 2444 2450]. This modeled complex relies primarily on the formation of five ion pairs between lysine residues of the cytochrome and acidic residues surrounding the flavin mononucleotide cofactor of the flavodoxin. In this study, the role of several acidic residues of the flavodoxin in the formation of this complex and in electron transfer between these two proteins was evaluated. A total of 17 flavodoxin mutants were studied in which 10 acidic amino acids--Asp62, Asp63, Glu66, Asp69, Asp70, Asp95, Glu99, Asp106, Asp127, and Asp129--had been permanently neutralized either individually or in various combinations by substitution with their amide amino acid equivalent (i.e., asparate to asparagine, glutamate to glutamine) through site-directed mutagenesis. The kinetic data for the transfer of electrons from reduced cytochrome c3 to the various flavodoxin mutants do not conform well to a simple bimolecular mechanism involving the formation of an intermediate electron transfer complex. Instead, a minimal electron transfer mechanism is proposed in which an initial complex is formed that is stabilized by intermolecular electrostatic interactions but is relatively inefficient in terms of electron transfer. This step is followed by a rate-limiting reorganization of that complex leading to efficient electron transfer. The apparent rate of this reorganization step was enhanced by the disruption of the initial electrostatic interactions through the neutralization of certain acidic amino acid residues leading to faster overall observed electron transfer rates at low ionic strengths. Of the five acidic residues involved in ion pairing in the modeled complex proposed by Stewart et al. (1988), the kinetic data strongly implicate Asp62, Glu66, and Asp95 in the formation of the electrostatic interactions that control electron transfer. Less certainty is provided by this study for the involvement of Asp69 and Asp129, although the data do not exclude their participation. It was not possible to determine whether the modeled complex represents the optimal configuration for electron transfer obtained after the reorganization step or actually represents the initial complex. The data do provide evidence for the importance of electrostatic interactions in electron transfer between these two proteins and for the existence of alternative binding modes involving acidic residues on the surface of the flavodoxin other than those proposed in that model. PMID- 9354633 TI - Rigidity of human alpha-fetoprotein tertiary structure is under ligand control. AB - Comparative study of the natural ligand effect on structural properties and conformational stability of human alpha-fetoprotein (AFP) and its homologue, human serum albumin (HSA), was performed using several approaches, including circular dichroism, fluorescence spectroscopy, and scanning microcalorimetry. Here we show that denaturation of AFP, induced by the increase of temperature or urea concentration, is irreversible. We have established the fact that this irreversibility is caused by ligand release from the AFP molecule. Interestingly, the ligand-free form of AFP has no rigid tertiary structure but exhibits substantial secondary structure and high compactness. This means that the rigid tertiary structure of AFP is controlled by interaction with ligands, while their release results in transition of a protein molecule into a molten globule-like intermediate. In contrast, processes of HSA denaturation and unfolding are completely reversible. Release of ligands from HSA results only in a small decrease in stability but not transformation into the molten globule state. PMID- 9354632 TI - Characterization of the unbound 2[Fe4S4]-ferredoxin-like photosystem I subunit PsaC from the Cyanobacterium synechococcus elongatus. AB - Recombinant PsaC was reconstituted in vitro and investigated by UV/vis, EPR, and 1H NMR spectroscopy. Its UV/vis and EPR spectroscopic properties correspond to those of the wild-type protein. Fast repetition 1D and 2D 1H NMR spectra allowed the sequence-specific assignment of the hyperfine-shifted proton resonances of the cluster-ligating resonances, taking advantage also of chemical shift analogies with other 4 and 8 Fe ferredoxins and a structural model for PsaC. The Calpha-Cbeta-S-Fe dihedral angles of the cluster ligands could be estimated from the chemical shifts and relaxation properties of their betaCH2 protons. All NMR derived structural information on PsaC confirms its similarity to smaller 8Fe ferredoxins serving as electron transfer proteins in solution. Partial reduction of PsaC leads to an intermediate species with strongly exchange broadened 1H NMR resonances. The intermolecular electron exchange rate is estimated to be in the 10(2)-10(4) s-1 range, the intramolecular electron exchange rate between the two [Fe4S4] clusters to be higher than 10(4) s-1. The consequences of these findings for the electron transfer in photosystem I are discussed. PMID- 9354634 TI - Domains in cationic lipid plus polyelectrolyte bilayer membranes: detection and characterization via 2H nuclear magnetic resonance. AB - 2H nuclear magnetic resonance (NMR) spectroscopy of choline-deuterolabeled 1 palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC-alpha-d2 and POPC-beta-d2) has been used to detect and quantify domain formation induced in cationic lipid containing bilayers upon the addition of anionic polyelectrolytes. Three different polyelectrolytes, poly(sodium 4-styrenesulfonate) or PSSS, poly(sodium acrylate) or PACA, and poly(sodium glutamate) or PGLU, were added to POPC lipid bilayers containing 1,2-dioleoyl-3-(dimethylamino)propane (DODAP) as the cationic amphiphile. All three polyelectrolytes produced two-component 2H NMR spectra, consistent with two populations of POPC, one polyelectrolyte-bound and another polyelectrolyte-free. The relative intensities of the two spectral components provided the relative amounts of the two POPC populations. The 2H NMR quadrupolar splitting from either spectral component provided the DODAP content of the particular POPC population. The two POPC populations differed in that the polyelectrolyte-bound population contained a stoichiometric polyelectrolyte anion:DODAP cation ratio leading to enrichment with respect to DODAP, while the polyelectrolyte-free population was depleted of DODAP. Estimates of the size of a polyelectrolyte-defined domain revealed a constant number of bound DODAP but a flexible number of bound POPC, which increased in proportion to the global POPC content. The most compact domains were formed by the most hydrophobic polyelectrolyte, PSSS, while the most expansive domains were formed by the most hydrophilic polyelectrolyte, PGLU. PMID- 9354635 TI - The helix-hinge-helix structural motif in human apolipoprotein A-I determined by NMR spectroscopy. AB - The conformation of a synthetic peptide of 46 residues from apoA-I was investigated by fluorescence, CD, and 2D NMR spectroscopies in lipid-mimetic environments. ApoA-I(142-187) is mainly unstructured in water but helical in SDS or dodecylphosphocholine (DPC), although the peptide only associates with DPC at approximately the critical micellar concentration. Solution structures of apoA I(142-187) were determined by distance geometry calculations based on 450 (in DPC d38) or 397 (in SDS-d25) NOE-derived distance restraints, respectively. Backbone RMSDs for superimposing the two helical regions 146-162 and 168-182 are 0.98 +/- 0.22 (2.38 +/- 0.20) and 1.99 +/- 0.42 (2.02 +/- 0.21) A in DPC (SDS), respectively. No interhelical NOE was found, suggesting that helix-helix interactions between the two helical domains in apoA-I(142-187) are unlikely. Similar average, curved helix-hinge-helix structures were found in both SDS and DPC micelles with the hydrophobic residues occupying the concave face, indicating that hydrophobic interactions dominate. Intermolecular NOESY experiments, performed in the presence of 50% protonated SDS, confirm that the two amphipathic helices and Y166 in the hinge all interact with the micelle. The involvement of Y166 in lipid binding is supported by fluorescence spectroscopy as well. On the basis of all the data above, we propose a model for the peptide-lipid complexes wherein the curved amphipathic helix-hinge-helix structural motif straddles the micelle. The peptide-aided signal assignment achieved for apoA-I(122-187) (66mer) and apoA-I suggests that such a structural motif is retained in the longer peptide and most likely in the intact protein. PMID- 9354636 TI - The Na/Ca-K exchanger of rod photoreceptor exists as dimer in the plasma membrane. AB - The oligomeric state of the Na/Ca-K exchanger in the plasma membrane of bovine photoreceptors was investigated using chemical cross-linking techniques. In the natural membrane, virtually all Na/Ca-K exchanger could be cross-linked mainly to a complex having an apparent molecular mass of 490 kDa by cupric phenanthroline catalyzed disulfide bonding as evidenced by Western blotting. Stable cross-links of the exchanger were also obtained with the thiol-specific reagent N,N'-p phenylidenedimaleimide. Neuraminidase treatment reduced the apparent molecular mass of the highly glycosylated Na/Ca-K exchanger and of the 490 kDa cross-link product by 50 and 85 kDa, respectively. DL-1,4-Bismaleimido-2,3-butanediol (BMBD), a novel cleavable dimaleimide, was synthesized in order to produce cross links that were stable to reductive conditions. Purification of the BMBD cross linked exchanger followed by two-dimensional SDS polyacrylamide electrophoresis identified the cross-linked homodimers of the exchanger. There was no indication of higher oligomers, suggesting that the exchanger exists as a dimer in the plasma membrane. Hydrodynamic properties of the detergent-solubilized exchanger were determined by velocity sedimentation and gel filtration chromatography. The Triton X-100-solubilized exchanger ran as a single species having a Stokes radius of 10.0 nm, a sedimentation coefficient of 5.4 S, and a partial specific volume of 0.74 mL/g in Triton X-100. Similar results were obtained for the CHAPS solubilized exchanger. A molecular mass of 236 and 205 kDa was calculated for the exchanger-detergent complex and the detergent-free protein, respectively. Neuraminidase treatment further reduced the molecular mass of the exchanger indicating that glycosylation contributes significantly to the mass of the exchanger. Cross-links of the exchanger were not detected if cross-linking was attempted after solubilization in 10 mM CHAPS. However, after reconstitution of the purified exchanger into soybean phosphatidylcholine vesicles, chemical cross linking yielded again dimers. On the basis of these cross-linking and hydrodynamic studies, we conclude that the exchanger exists as a homodimer in the rod outer segment plasma membrane but dissociates into a monomer when solubilized in detergent. PMID- 9354637 TI - Nitric oxide modulates the c-Jun N-terminal kinase/stress-activated protein kinase activity through activating c-Jun N-terminal kinase kinase. AB - Nitric oxide is a signaling molecule that has a broad range of physiological functions, including neurotransmission, macrophage activation, and vasodilation. The mechanism by which nitric oxide regulates signal transduction mediating diverse biological activities is not fully understood, however. Here, we demonstrate that nitric oxide induced the stimulation of c-Jun NH2-terminal kinase (JNK)/stress-activated protein kinase (SAPK) in intact cells. Exposure of cultured HEK293 cells to sodium nitroprusside, a nitric oxide releasing agent, resulted in the stimulation of JNK1 activity. The sodium nitroprusside-induced stimulation of JNK1 activity was abolished by treatment of cells with N acetylcysteine. Nitric oxide production from HEK293 cells ectopically expressing nitric oxide synthases resulted in the stimulation of JNK1 activity, while JNK1 stimulation in nitric oxide synthase-overexpressing cells was abrogated by a nitric oxide synthase inhibitor, NG-nitro-L-arginine. Furthermore, exposure of cells to sodium nitroprusside resulted in the stimulation of JNK kinase (JNKK1/SEK1). Taken together, our data suggest that nitric oxide modulates the JNK activity through activating JNKK1/SEK1. PMID- 9354638 TI - Arginine 302 (helix IX) in the lactose permease of Escherichia coli is in close proximity to glutamate 269 (helix VIII) as well as glutamate 325. AB - By using a variety of biochemical and biophysical approaches, a helix packing model for the lactose permease of Escherichia coli has been proposed in which the four residues that are irreplaceable with respect to coupling are paired--Glu269 (helix VIII) with His322 (helix X) and Arg302 (helix XI) with Glu325 (helix X). In addition, the substrate translocation pathway is located at the interface between helices V and VIII, which is in close vicinity to the four essential residues. Based on this structural information and functional studies of mutants in the four irreplaceable residues, a molecular mechanism for energy coupling in the permease has been proposed [Kaback, H. R. (1997) Pro.c Natl. Acad. Sci. U.S.A. 94, 5539]. The principle idea of this model is that Arg302 interacts with either Glu325 or Glu269 during turnover. Evidence that Arg302 is in close proximity with Glu325 has been presented [Jung, K., Jung, H., Wu, J., Prive, G. G., & Kaback, H. R. (1993) Biochemistry 32, 12273; He, M. M., Voss, J., Hubbell, W. L., & Kaback, H. R. (1995) Biochemistry 34, 15667]; however, the proximity of Arg302 to Glu269 has not been examined. In this report, it is shown by two methods that Arg302 is also close to Glu269: (i) permease with Glu269-->His, Arg302-->His, and His322-->Phe binds Mn2+ with high affinity at pH 7.5, but not at pH 5.5; and (ii) site-directed spin-labeling of the double Cys mutant Glu269- >Cys/Arg302-->Cys exhibits spin-spin interaction with an interspin distance of about 14-16 A. In addition, the spin-spin interaction is stronger and interspin distance shorter after the permease is reconstituted into proteoliposomes. Taken as a whole, the data are consistent with the idea that Arg302 may interact with either Glu325 or Glu269 during turnover. PMID- 9354639 TI - Interaction between residues Glu269 (helix VIII) and His322 (helix X) of the lactose permease of Escherichia coli is essential for substrate binding. AB - Site-directed and Cys-scanning mutagenesis of the lactose permease of Escherichia coli reveals that as few as four residues--Glu269 (helix VIII), Arg302 (helix IV), His322 (helix X), and Glu325 (helix X)--are irreplaceable for coupling substrate and H+ translocation. Interestingly, the four residues are in close physical proximity, Glu269 interacting with His322 and Arg302 with Glu325. In addition, the substrate translocation pathway is located close to the four residues at the interface between helices V and VIII. To investigate the importance of the four residues and their interactions for substrate binding, mutation Glu269-->Asp, Glu269-->Gln, Arg302-->Ala, Arg302-->Lys, His322-->Ala, His322-->Phe, Glu325-->Asp, or Glu325-->Gln was introduced into single-Cys148 permease, where the reactivity of Cys with 2-(4-maleimidoanilino)naphthalene-6 sulfonic acid (MIANS) is blocked by binding of substrate. The double mutants were purified, and the rates of MIANS labeling were measured in the absence or presence of beta-D-galactopyranosyl 1-thio-beta-D-galactopyranoside (TDG), lactose, or galactose at various concentrations. Remarkably, substrate binding by the Glu269 or His322 mutants is abolished or decreased dramatically, while binding by the Arg302 or Glu325 mutants is not altered. The observations are consistent with the notion that the interaction between Glu269 and His322 stabilizes the interface between helices V and VIII and thereby leads to binding of substrate. PMID- 9354640 TI - A single amino acid substitution in ribonucleolytic toxin restrictocin abolishes its specific substrate recognition activity. AB - Restrictocin is a small basic protein produced by the fungus Aspergillus restrictus. It potently inhibits protein synthesis in eukaryotic cells by specifically cleaving a single phosphodiester bond in 28S rRNA. A histidine residue at position 49 in restrictocin has been implicated in its active site. A mutant of restrictocin in which the histidine at position 49 was changed to an alanine was constructed by site-directed mutagenesis, and the protein was expressed in Escherichia coli. The mutant and the wild type proteins were found to be structurally identical. Unlike restrictocin, the restrictocin H49A mutant did not cleave the ribosomal RNA specifically at the target phosphodiester bond; instead, it extensively degraded the RNA substrate with altered specificity. The mutant exhibited a high ribonuclease activity compared to restrictocin on yeast tRNA, and poly(U) and poly(C). The mutant also poorly inhibited protein synthesis in eukaryotic cells as well as in a cell free system. We therefore propose that histidine 49 of restrictocin is not involved per se in the enzymatic activity; however, it does play a crucial role in the specific recognition of the target sequence by restrictocin. PMID- 9354641 TI - Conformational analysis of Escherichia coli 30S ribosomes containing the single base mutations G530U, U1498G, G1401C, and C1501G and the double-base mutation G1401C/C1501G. AB - Biochemical and genetic studies have pointed out the importance of several sites in 16S ribosomal RNA of Escherichia coli in the decoding process. These sites consist of the core of the decoding center (1400/1500 region) and two other segments (530 and 1050/1200 regions). To detect a possible structural link between these functionally related regions, we analyzed their sensitivity to conformational changes induced by mutations which are located in each of these regions and are known to affect the decoding process. The conformations of five segments of 16S rRNA (1-106, 406-569, 780-978, 997-1247, and 1334-1519) were analyzed by chemical probing of 30S ribosomes containing the following mutations: G530U, U1498G, G1401C, C1501G, and G1401C/C1501G. Ribosomes reconstituted with natural wild-type 16S RNA showed only minor conformational differences with respect to ribosomes isolated from cells. When 16S RNA made in vitro replaced natural 16S RNA, a slightly looser conformation of the central core region was found. Mutant ribosomes made by reconstitution with mutant 16S RNA made in vitro showed conformational effects which were in all cases localized to the region of secondary structure surrounding the site of mutation. Although the core of the decoding center (1400/1500 region) and the two other sites (530 and 1050/1200 regions) participating in the decoding function have been functionally linked, our data indicate that they are structurally independent. They also provide evidence for an unusual structure of the 1400/1500 decoding center, possibly involving noncanonical interactions. Furthermore, the absence of any conformational effect induced by the G530U mutation except at the site of mutation itself points to its direct, as opposed to indirect, involvement in the decoding function of the ribosome. PMID- 9354642 TI - Strand-specific modulation of UV photoproducts in 5S rDNA by TFIIIA binding and their effect on TFIIIA complex formation. AB - The relationship between UV-induced photoproduct formation and transcription factor binding was studied in a 214 bp fragment containing the entire Xenopus borealis 5S rRNA gene. DNA mobility shift and DNase I footprinting show a strong inhibition of TFIIIA binding to UV-damaged 5S rDNA. An average of approximately 2 cyclobutane pyrimidine dimers (CPDs) per 214 bp fragment, and a lesser amount of pyrimidine-pyrimidone (6-4) dimers, reduced the fraction of TFIIIA bound by approximately 70%. Furthermore, irradiation of the TFIIIA/5S rDNA complex displaces TFIIIA at doses of 0.8-2 CPDs/fragment, indicating the complex is unable to accommodate UV photoproducts. UV photofootprinting of the 50 bp TFIIIA binding region of 5S rDNA (or ICR) shows that TFIIIA binding modulates photoproduct formation primarily in the template strand. Formation of CPDs at six different sites is strongly inhibited, while another CPD site is strongly enhanced, by TFIIIA binding. Most of these sites are located in one of three boxes (A, IE, or C) designated as TFIIIA contact sites in the ICR, while one site is between these boxes. Formation of (6-4) dimers is also inhibited at several sites in the template strand by TFIIIA binding. However, formation of photoproducts in the nontemplate strand is much less affected by TFIIIA binding, where only one CPD site is inhibited in the complex. These data indicate that formation of UV photoproducts in 5S rDNA can be markedly affected by TFIIIA binding, and complex formation is inhibited by UV photoproducts. PMID- 9354643 TI - Determinants of ribose specificity in RNA polymerization: effects of Mn2+ and deoxynucleoside monophosphate incorporation into transcripts. AB - The catalytic specificity of T7 RNA polymerase (RNAP) for ribonucleoside triphosphates vs deoxynucleoside triphosphates {(kcat/Km)rNTP/(kcat/Km)dNTP} during transcript elongation is approximately 80. Mutation of tyrosine 639 to phenylalanine reduces specificity by a factor of approximately 20 and largely eliminates the Km difference between rNTPs and dNTPs. The remaining specificity factor of approximately 4 is kcat-mediated and is nearly eliminated if Mn2+ is substituted for Mg2+ in the reaction. Mn2+ substitution does not significantly affect the Km difference between rNTPs and dNTPs. Mn2+ substitution also enhances the activity of poorly active mutant enzymes carrying nonconservative substitutions in the active site, and its effects are generally consistent with the Mn2+-catalyzed reaction being less restrictive in its requirements for alignment of the reactive groups. In addition to discrimination occurring at the level of nucleoside monophosphate (NMP) incorporation, it is also found that transcripts containing deoxynucleoside monophosphates (dNMPs) are more poorly extended than transcripts of canonical structure, though a severe barrier to transcript extension is seen only when the 3' region of the transcript is heavily substituted with dNMPs. The barrier to extension of transcripts heavily substituted with dNMPs is reduced for sequences known to be amenable to forming A like helices and is larger for sequences that resist transformation from B-form DNA.DNA structures. The barrier to extension of dNMP-substituted transcripts is also reduced by solution conditions known to destabilize B-form DNA and to stabilize A-form structures. These observations imply a requirement for a non-B form, possibly A-like, conformation in the transcript.template hybrid that is disrupted when the transcript is of predominantly deoxyribose structure. PMID- 9354645 TI - A conserved glutamic acid in helix VI of cytochrome bo3 influences a key step in oxygen reduction. AB - We have compared the reactions with dioxygen of wild-type cytochrome bo3 and a mutant in which a conserved glutamic acid at position-286 of subunit I has been changed to an alanine. Flow-flash experiments reveal that oxygen binding and the rate of heme-heme electron transfer are unaffected by the mutation. Reaction of the fully (3-electron) reduced mutant cytochrome bo3 with dioxygen yields a binuclear center which is substantially in the P (peroxy) state, not the well characterized F (oxyferryl) state which is the product of the reaction of the fully reduced wild-type enzyme with dioxygen [Puustinen, A., et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 1545-1548]. These results confirm that proton uptake is important in controlling the later stages of dioxygen reduction in heme-copper oxidases and show that E286 is an important component of the channel that delivers these protons to the active site. PMID- 9354644 TI - Enzymatic midchain branching of polylactosamine backbones is restricted in a site specific manner in alpha 1,3-fucosylated chains. AB - Branched polylactosamines on animal cell surfaces are believed to contribute to multivalent interactions in cell adhesion and cell signalling. Their biosynthesis proceeds via linear precursors that become branched by beta1,6-GlcNAc transferases (IGnT6, GlcNAc to Gal). Previous work has identified the tetrasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAc (1) and the hexasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1-4GlcNAc (4) as acceptors for a rat serum enzyme activity (cIGnT6), which transfers GlcNAcbeta1-6 units to the midchain galactose residues. Thereby, 1 is converted to the branched pentasaccharide Galbeta1-4GlcNAcbeta1-3(GlcNAcbeta1-6)Galbeta1-4 GlcNAc and 4 to the doubly branched octasaccharide Galbeta1-4GlcNAcbeta1 3(GlcNAcbeta1-6)Galbeta1-+ ++4GlcNAcbeta1-3(GlcNAcb eta1-6)Galbeta1-4GlcNAc [Leppanen, A., Salminen, H., Zhu, Y., Maaheimo, H., Helin, J., Costello, C. E., & Renkonen, O. (1997) Biochemistry 36, 7026-7036]. Here we report that neither the alpha1, 3-fucose-containing derivatives of 1 [Galbeta1-4GlcNAcbeta1-3Galbeta1 4(Fucalpha1-3)G lcNAc and Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4Gl cNAc] nor a similar derivative of 4 [Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)+ ++GlcNAcbeta1-3Galbeta1- 4GlcNAc] were acceptors for the rat serum cIGnT6 activity. Hence, the enzyme's branch-forming action was completely prevented at sites in the immediate neighborhood of the fucosylated loci of the polylactosamines. In Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1 4(Fucalpha1-3) GlcNAc, the inhibition of the branch-forming reaction was restricted to the fucose-carrying LacNAc unit; at the middle LacNAc, the branching proceeded normally. However, in the isomeric Galbeta1-4(Fucalpha1 3)GlcNAcbeta1-3Galbeta1- 4GlcNAcbeta1-3Galbeta1-4 GlcNAc, the fucose residue prevented branching completely at the middle LacNAc and almost completely at the reducing end LacNAc. In summary, alpha1,3-fucose residues in polylactosamine chains inhibited the cIGnT6 reaction in a site-specific manner, at the fucosylated LacNAc unit itself and also at sites one and two LacNAc units upstream, but not at the LacNAc units downstream from the fucosylated locus. These data imply that site-directed branching in polylactosamines is possible in vitro with the aid of specifically positioned alpha1,3-fucosyl units, that can be removed afterward without harming the branched backbones. PMID- 9354646 TI - Expression and mutagenesis of mammalian cytosolic NADP+-specific isocitrate dehydrogenase. AB - Rat liver cytosolic NADP+-specific isocitrate dehydrogenase (IDP2) was expressed in bacteria as a fusion protein with maltose binding protein (MBP). High levels of expression were obtained. The fusion protein was purified from bacterial lysates by affinity chromatography with an amylose resin and found to be catalytically active. IDP2 was separated from MBP by cleavage with protease Xa and purified to homogeneity by FPLC anion-exchange chromatography. A specific activity of 56.3 units/mg and respective apparent Km values for dl-isocitrate and NADP+ of 9.7 +/- 2.9 microM and 11.5 +/- 0.2 microM were obtained for the purified enzyme. These values are similar to those previously reported for cytosolic isocitrate dehydrogenase isolated from a variety of tissues. Evolutionarily conserved arginine residues implicated in substrate binding were changed to glutamate residues using PCR based site-directed mutagenesis of the bacterial fusion plasmid. Mutant enzymes containing residue changes of R100E, R109E, R119E, or R132E were expressed, purified, and characterized by initial rate kinetic analyses. The R119E and R109E mutant enzymes exhibited respective 15 and 31-fold increases in Km values for dl-isocitrate relative to the wild-type enzyme. In contrast, Km values for NADP+ were, respectively, unchanged and increased 9-fold. The most significant reductions in kcat/Km values were obtained for the R100E, R109E, and R132E enzymes. These results suggest that substrate binding residues are highly conserved between bacterial and mammalian enzymes despite low overall homology. PMID- 9354647 TI - Inhibition of recombinant human mitochondrial and cytosolic aldehyde dehydrogenases by two candidates for the active metabolites of disulfiram. AB - We expressed recombinant human cytosolic (ALDH1, high Km) and mitochondrial aldehyde dehydrogenase (ALDH2, low Km) in Escherichia coli and purified the enzymes to homogeneity to examine the nature of inhibition of human ALDH by disulfiram, its confirmed metabolite S-methyl N,N-diethylthiocarbamate (MeDTC) sulfoxide, and its proposed metabolite MeDTC sulfone. Disulfiram, MeDTC sulfoxide, and MeDTC sulfone, respectively, were potent inhibitors with IC50 values of 0.15 +/- 0.02 microM, 0.27 +/- 0.04 microM, and 0.12 +/- 0.02 microM for ALDH1, and 1.45 +/- 0.40 microM, 1.16 +/- 0.56, and 0.40 +/- 0.10 microM for ALDH2. Extensive dialysis did not restore the activity of the inactivated enzyme, indicating irreversible inhibition. Both the esterase and dehydrogenase activities of ALDH2 were inhibited to the same extent by MeDTC sulfone and sulfoxide, suggesting that both catalytic sites are closely linked. The time course of inhibition of ALDH appeared to be first-order for both MeDTC sulfone and MeDTC sulfoxide. Kitz and Wilson plots of the half-life of inactivation versus 1/[inhibitor] indicated that the reactions between ALDH and inhibitors were bimolecular. The pseudobimolecular rate constants (k3/KI) for the ALDH inhibitor reactions were 1 x 10(5), 1 x 10(4), 3 x 10(3), and 1 x 10(3) s-1 M-1 ALDH1-sulfone, ALDH1-sulfoxide, ALDH2-sulfone, and ALDH2-sulfoxide, respectively. ALDH2 was not significantly protected from inactivation from either MeDTC sulfoxide or MeDTC sulfone by NAD alone, but high concentrations of NAD and acetaldehyde completely prevented inhibition. Since disulfiram is rapidly metabolized in vivo, it is believed that disulfiram is too short-lived to inhibit ALDH directly. The results of our study indicate that MeDTC sulfoxide and sulfone are potent inhibitors of human ALDH and are reasonable candidates for the proximal inhibitors of ALDH following disulfiram administration. PMID- 9354648 TI - IgG biosynthesis: no "immunoregulatory feedback". PMID- 9354649 TI - Contact system: a vascular biology modulator with anticoagulant, profibrinolytic, antiadhesive, and proinflammatory attributes. PMID- 9354650 TI - Results of high-dose therapy and autologous bone marrow/stem cell transplantation during remission in poor-risk intermediate- and high-grade lymphoma: international index high and high-intermediate risk group. AB - We have conducted a pilot study to investigate the role of high-dose therapy and autologous bone marrow/stem cell transplantation (ASCT) during first complete or partial remission in 52 patients with poor-risk aggressive lymphoma. There were 42 patients with intermediate-grade or immunoblastic lymphoma who were considered to be high (60%) and high-intermediate risk (40%) groups at diagnosis based on the age-adjusted International Prognostic Index (IPI) and 10 patients with high grade, SNCCL (small non-cleaved cell, Burkitt's, and non-Burkitt's), who at presentation had poor-risk features defined as elevated serum lactate dehydrogenase level, stage IV, and bulky mass >/=10 cm. The median age was 34 years (range, 16 to 56 years). Thirty-nine were transplanted in first complete remission and 13 in first partial remission after conventional therapy. Conditioning regimens consisted of total body irradiation (TBI) administered as a single fraction 750 cGy in 3 patients and in fractionated doses for a total of 1,200 cGy in 44 patients, in combination with 60 mg/kg etoposide and 100 mg/kg cyclophosphamide. Five patients with prior radiotherapy received 450 mg/m2 carmustine instead of TBI. Stem cell sources were either bone marrow and/or peripheral blood. No in vitro purging was used. All patients engrafted. Two SNCCL patients died of venoocclusive disease at 25 days and acute leukemia at 27 months posttransplantation. There were six relapses at 1.5 to 12.8 months posttransplantation. At a median follow-up of 44 months (range, 1 to 113 months), the estimated 3-year overall survival (OS) and disease-free survival (DFS) for all patients was 84% (95% confidence interval [CI], 70% to 92%) and 82% (95% CI, 68% to 91%), respectively. In the subset of patients with intermediate-grade and immunoblastic lymphoma, the 3-year DFS was 89% (95% CI, 74% to 96%) for all patients, 87% (95% CI, 67% to 96%) for high-risk patients, and 92% (95 CI, 61% to 99%) for high-intermediate risk patients. The 3-year OS and DFS for SNCCL patients were identical at 60% (95% CI, 30% to 84%). These results suggest that high-dose therapy and ASCT during first remission may improve the survival and prognosis of patients with poor-risk intermediate- and high-grade lymphoma. A prospective randomized study comparing high-dose therapy and ASCT with conventional chemotherapy in IPI high-risk patients with aggressive non-Hodgkin's lymphoma should be undertaken. PMID- 9354652 TI - Hepatic lesions of chronic disseminated candidiasis may become invisible during neutropenia. AB - We describe the phenomenon of waning of focal hepatic and/or splenic lesions on abdominal computed tomographic (CT) scan during neutropenia in patients with chronic disseminated candidiasis. After observation of the phenomenon in one patient, a total of five cases were prospectively monitored with serial CT scans. After the diagnosis of disseminated candidiasis, hepatic lesions decreased in size and conspicuousness in three patients, while in two others they disappeared completely during a subsequent chemotherapy-induced neutropenia. After recovery of the neutrophils, the lesions reappeared or increased in conspicuousness in all five patients. Of three patients treated with a second cycle of myeloablative chemotherapy, lesions again decreased in two patients during neutropenia and increased again in one patient after neutrophil recovery. In all five patients, candidiasis eventually resolved after prolonged antifungal treatment. In chronic disseminated candidiasis, hepatic or splenic lesions may transiently disappear during neutropenia. Thus, antifungal therapy should not be discontinued on the basis of radiologic findings alone. PMID- 9354651 TI - Autologous bone marrow transplantation for patients with myelodysplastic syndrome (MDS) or acute myeloid leukemia following MDS. Chronic and Acute Leukemia Working Parties of the European Group for Blood and Marrow Transplantation. AB - Intensive chemotherapy followed by autologous bone marrow transplantation (ABMT) may provide an alternative therapy for young patients with myelodysplastic syndrome (MDS) or acute myeloid leukemia following MDS (sAML) lacking a suitable donor. We report the results for 79 patients with MDS/sAML transplanted with autologous marrow in first complete remission (CR). Within the total group of 79, a cohort of 55 patients for whom the duration of first CR was known were compared with a matched control group of 110 patients with de novo AML. The 2-year survival, disease-free survival (DFS), and relapse rates for the 79 patients transplanted in first CR were 39%, 34%, and 64%, respectively. The relapse risk was greater than 55% for all stages and all disease categories. Patients younger than 40 years had a significantly (P = .04) better DFS (39%) than patients older than 40 years (25%). The DFS at 2 years was 28% for the cohort of 55 patients transplanted for MDS/sAML and 51% for those transplanted for de novo AML (P = .025). Relapse rates were 69% for patients with MDS/sAML and 40% for those with de novo AML (P = .007). ABMT for MDS or secondary leukemia results in a lower DFS when compared with similarly treated patients with de novo AML due to a higher relapse rate. The DFS of 28% for these patients suggests that autotransplantation may be a valuable therapy for this disease. The low treatment-related mortality rate of less than 10% supports the view that sufficient numbers of hematopoietic stem cells are present in patients with MDS to allow adequate repopulation after autologous stem-cell transplantation. PMID- 9354653 TI - Predictors of long-term response to high-dose interferon therapy in type II cryoglobulinemia associated with hepatitis C virus infection. AB - We have prospectively studied patients with type II cryoglobulinemia since 1985 to assess the efficacy of treatment with interferon-alpha at cumulative doses ranging from 234 to 849 MU. In the present study we retrospectively evaluated in this cohort parameters associated with complete response to therapy in 31 consecutive patients with type II cryoglobulinemia associated with hepatitis C virus (HCV) infection. Prevalence of complete response of cryoglobulinemia (disappearance of symptoms and signs of vasculitis and decrease of cryocrit below 10% of the initial value) was 62%, with a median response duration of 33 months and a range of 3 to 100 months. Three patients were putatively cured, as they remained in complete remission for more than 5 years off therapy. Eighteen patients (58%) had liver disease evidenced by histopathology and/or raised transaminase levels. Prevalence of normalization of transaminase levels was 100%, with a median response duration of 36 months. Relapse of hypertransaminasemia occurred in 100% and 8% of patients receiving less than or greater than 621 MU, respectively. By logistic regression analysis, the only pretherapy parameter that associated significantly (P = .0393) with complete response of cryoglobulinemia was the solitary anti-C22 (HCV core) antibody pattern, which was observed in 29% of patients. Association with older age and low cryocrit approached statistical significance (P = . 06), while no significant correlations were found with serum IgM levels, duration of disease, HCV genotype, NS5a gene mutations, liver histology, HLA-DR phenotype, or WA cross-idiotype. Complete responses were also associated, on univariate statistical analysis, with low pretherapy HCV viremia. Responses were accompanied by decrease of viremia, of anti-HCV antibody levels and cryocrit. The usefulness of a high dose regimen is underscored by the higher rates of sustained responses of cryoglobulinemia and transaminase levels compared with previous studies. PMID- 9354654 TI - Increased growth promoting but not mast cell degranulation potential of a covalent dimer of c-Kit ligand. AB - The native form of soluble c-kit ligand (KL) is a noncovalent dimer. We have isolated a soluble, disulfide-linked dimer of murine KL (KL-CD) by expressing KL in Escherichia coli and refolding the denatured protein under conditions that promote the formation of both noncovalent dimers (KL-NC) and KL-CD. KL-CD exhibits a 10- to 15-fold increase in the ability to stimulate the growth of both the human megakaryocytic cell line MO7e and murine bone marrow-derived mast cells relative to KL-NC. Colony-forming assays of murine bone marrow progenitor cells also reflected this increased potency. However, KL-CD and KL-NC are equally able to prime mast cells for enhanced IgE-dependent degranulation in vitro and activate mast cells in vivo. Improving the growth-promoting activity of KL without changing its mast cell activation potential suggests that KL-CD or a related molecule could be administered in the clinic at doses that stimulate hematopoietic recovery while avoiding significant mast cell activation. PMID- 9354655 TI - Development of a novel selective amplifier gene for controllable expansion of transduced hematopoietic cells. AB - To overcome the low efficiency of gene transfer into hematopoietic cells, we developed a novel system for selective expansion of transduced cells. To this end, we constructed a chimeric cDNA (GCRER) encoding the fusion protein between the granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone binding domain (HBD) of the estrogen receptor (ER) as a selective amplifier gene. Use of the intracellular signaling pathway of G-CSFR was considered to be appropriate, because G-CSF has the ability not only to stimulate the neutrophil production, but also to expand the hematopoietic stem/progenitor cell pool in vivo. To activate the exogenous G-CSFR signal domain selectively, the estrogen/ER HBD system was used as a molecular switch in this study. When the GCRER gene was expressed in the interleukin-3 (IL-3)-dependent murine cell line, Ba/F3, the cells showed IL-3-independent growth in response to G-CSF or estrogen. Moreover, the Ba/F3 cells transfected with the Delta(5-195)GCRER, whose product lacks the extracellular G-CSF-binding domain, did not respond to G-CSF, but retained the ability for estrogen-dependent growth. Further, murine bone marrow cells transduced with the GCRER or Delta(5-195)GCRER gene with retroviral vectors formed a significant number of colonies in response to estrogen, as well as G CSF, whereas estrogen did not stimulate colony formation by untransduced murine bone marrow cells. It is noteworthy that erythroid colonies were apparently formed by the bone marrow cells transduced with the GCRER gene in the presence of estrogen without the addition of erythropoietin, suggesting that the signals from the G-CSFR portion of the chimeric molecules do not preferentially induce neutrophilic differentiation, but just promote the differentiation depending on the nature of the target cells. We speculate that when the selective amplifier genes are expressed in the primitive hematopoietic stem cells, the growth signal predominates and that the population of transduced stem cells expands upon estrogen treatment, even if some of the cells enter the differentiation pathway. The present study suggests that this strategy is applicable to the in vivo selective expansion of transduced hematopoietic stem cells. PMID- 9354656 TI - Recombinant human interleukin-11 directly promotes megakaryocytopoiesis in vitro. AB - We have investigated the mechanism of action of the thrombopoietic cytokine, recombinant human interleukin-11 (rhIL-11), on megakaryocytopoiesis in vitro. We have shown that rhIL-11-induced murine and human megakaryocytopoiesis are not mediated by thrombopoietin (Tpo). Murine megakaryocytes (MKs) were produced from bone marrow (BM) mononuclear cells cultured with rhIL-11, IL-3, and a combination of the two cytokines. Conditioned media (CM) were collected and assayed for the presence of biologically active Tpo. Tpo activity was not detected in any of the CMs tested. Next, human BM CD34+ cells were cultured in serum-free fibrin clot medium with rhIL-11, IL-3, or rhIL-11 plus IL-3 and an antibody that neutralizes human Tpo activity. No inhibition of either burst-forming unit-MK- or colony forming unit-MK-derived colony formation was observed. The antibody did partially inhibit steel factor-induced MK-colony formation, suggesting that the actions of this cytokine are mediated, in part, by Tpo. We determined that MKs can be direct targets of rhIL-11 by showing the expression of functional IL-11 receptor on these cells. Total RNA was prepared from cultured human BM CD41+CD14- cells (MKs) and IL-11 receptor alpha chain mRNA was detected in the MKs by reverse transcription-polymerase chain reaction. Analysis of single-sorted CD41+CD14- cells confirmed that the observed IL-11 receptor expression was not due to contaminating CD41- cells in the pool. The presence of rhIL-11 receptor alpha chain protein in the cells was established by Western blot analysis. After a short exposure of purified BM MKs to rhIL-11, enhanced phosphorylation of both its signal transduction subunit, gp130, and the transcription factor, STAT3 was detected, showing a direct activation of receptor signaling by the cytokine. Consistent with the lack of effect of rhIL-11 on platelets in vivo, IL-11 receptor alpha chain mRNA and protein were not detected in isolated human platelets. These data indicate that rhIL-11 acts directly on MKs and MK progenitors but not on platelets. PMID- 9354657 TI - Flt3 ligand enhances the yield of primitive cells after Ex vivo cultivation of CD34+ CD38dim cells and CD34+ CD38dim CD33dim HLA-DR+ cells. AB - Flt3 ligand (FL) has been proposed as a possible modulator of early hematopoietic cell growth. The purpose of this study was to analyze the impact of FL on ex vivo expansion of hematopoietic cells obtained from adult donors. We sought to precisely identify hematopoietic populations responsive to FL and to quantitate the ability of FL to enhance the survival and/or proliferation of early hematopoietic precursors in a stroma-free culture system. Towards that end, four CD34+ subsets were isolated and their response to FL was characterized. In methylcellulose, FL significantly increased colony formation by CD34+ CD38dim cells but not CD34+ CD38+ cells. In suspension culture, the enhancement of cell expansion by FL was 10 times greater with the CD34+ CD38dim fraction than the CD34+ CD38+ fraction. FL stimulated the generation of colony-forming unit granulocyte-macrophage (CFU-GM) from the CD34+CD38dim fraction by 14.5- +/- 5.6 fold. To determine if CD34+ CD38dim cells responded uniformly to FL, the population was subdivided into a CD34+ CD38dim CD33dim HLA-DR+ (HLA-DR+) fraction and a CD34+ CD38dim CD33(dim) HLA-DRdim (HLA-DRdim) fraction. FL was far more effective at stimulating cell and progenitor growth from the HLA-DR+ fraction. To determine if FL enhanced or depleted the number of precommitted cells in expansion culture, CD34+ CD38dim and HLA-DR+ fractions were incubated in liquid culture and analyzed by flow cytometry. Inclusion of FL enhanced the absolute number of primitive CD34+ CD33dim cells and CD34+ HLA-DRdim cells after 5 to 12 days of cultivation. To confirm immunophenotypic data, the effect of FL on long term culture-initiating cells (LTCIC) was determined. After 2 weeks of incubation of CD34+ CD38dim or HLA-DR+ cultures, LTCIC recoveries were significantly higher with FL in 5 of 6 trials (P < . 05). For HLA-DR+ cells, LTCIC recoveries averaged 214% +/- 87% of input with FL and 24% +/- 16% without FL. In contrast, HLA-DRdim LTCIC could not be maintained in stroma-free culture. We conclude that less than 10% of CD34+ cells respond vigorously to FL and that those cells are contained within the HLA-DR+ fraction. FL stimulates the expansion of total cells, CD34+ cells, and CFU-GM and enhances the pool of early CD34+ CD33(dim) cells, CD34+ HLA DRdim cells, and LTCIC. These data indicate that it is possible to expand hematopoietic progenitors from adult donors without losing precursors from the precommitted cell pool. PMID- 9354659 TI - Development of natural killer cells, B lymphocytes, macrophages, and mast cells from single hematopoietic progenitors in culture of murine fetal liver cells. AB - We have recently established a clonal culture system that supports the growth of immature natural killer (NK) cells from murine fetal thymocytes. We now describe a culture system for mixed NK cell colony formation from single lymphohematopoietic progenitors. When Sca-1+c-kit+ fetal liver cells were cultured in methylcellulose media with interleukin (IL)-2, IL-7, IL-11, and steel factor (SF), we found mixed colonies consisting of diffuse small round cells characteristic of immature NK cells and other types of cells. The single cell origin of the mixed colonies was established by micromanipulation. Individual mixed colonies derived from single cells were characterized by flow cytometric analysis and May-Grunwald Giemsa staining. All mixed colonies contained Thy 1+B220- cells, which can differentiate to mature NK cells in fetal thymus organ culture. Most of the colonies contained B220+ B-lineage cells and macrophages, and some contained mast cells. IL-1alpha and IL-3, which have previously been shown to inhibit the T- and B-cell potentials of blast colonies, suppressed the formation of mixed NK cell colonies. The clonal culture assay presented here may be useful in analysis of the developmental pathway and commitment of NK cells from multipotential progenitors. PMID- 9354658 TI - Mitogenic responses mediated through the proteinase-activated receptor-2 are induced by expressed forms of mast cell alpha- or beta-tryptases. AB - The proteinase-activated receptor-2 (PAR-2) is the second member of a putative larger class of proteolytically activated receptors that mediate cell activation events by receptor cleavage or synthetic peptidomimetics corresponding to the newly generated N-terminus. To further study the previously identified mitogenic effects of PAR-2, we used the interleukin-3 (IL-3)-dependent murine lymphoid cell line, BaF3, for generation of stable cell lines expressing PAR-2 (BaF3/PAR-2) or the noncleavable PAR-2 mutant PAR-2(Arg36 --> Ala36). Only BaF3 cells expressing either wild-type or mutated receptor exhibited mitogenic responses when grown in IL-3-deficient media supplemented with PAR-2 activating peptide (SLIGRL, PAR39 44). This effect was dose dependent with an EC50 of approximately 80 micromol/L, sustained at 24, 48, and 72 hours, and was also demonstrable using thrombin receptor peptide TR42-47. Because tryptase shares approximately 70% homology with trypsin (previously shown to activate PAR-2), we studied recombinantly expressed forms of alpha- and beta-tryptases as candidate protease agonists for PAR-2. Hydrolytic activity of the chromogenic substrate tosyl-glycyl-prolyl-argly-4 nitroanilide acetate was present as a sharp peak at Mr approximately 130, confirming the presence of secretable and functionally active homotetrameric alpha- and beta-tryptases in transfected COS-1 cells. Dose-dependent proliferative responses were evident using either secreted form of tryptase with maximal responses seen at approximately 3 pmol/L (0.1 U/L). Receptor proteolysis was necessary and sufficient for mitogenesis because active site-blocked tryptase failed to induce this response, and proliferative responses were abrogated in BaF3 cells expressing PAR-2(Arg36 --> Ala36). These results specifically identify both forms of mast cell tryptases as serine protease agonists for PAR-2 and have implications for elucidating molecular mechanisms regulating cellular activation events mediated by proteases generated during inflammatory, fibrinolytic, or hemostatic-regulated pathways. PMID- 9354660 TI - Enhanced lipid peroxidation in patients positive for antiphospholipid antibodies. AB - The mechanism leading to the formation of antiphospholipid antibodies (aPL) is still unknown. Because an in vitro study suggested that aPL may derive from pro oxidant conditions, we sought a relationship between aPL and isoprostanes, indices of lipid peroxidation in vivo. Thirty patients with systemic lupus erythematosus have been studied. Seventeen (56.6%) were positive for aPL because they had lupus anticoagulant and/or high titer of anticardiolipin antibodies (aCL). Plasma levels of tumor necrosis factor (TNF ) and urinary excretion of two isoprostanes, 8-epi-PGF2alpha and IPF2alpha -I, free radical catalyzed oxidation products of arachidonic acid, were measured. Patients with systemic lupus erythematosus had higher urinary excretion of 8-epi-PGF2alpha and IPF2alpha -I than controls; urinary excretion of the two isoprostanes was highly correlated (Rho = 0.74, P < .0001). Urinary 8-epi-PGF2alpha was highly correlated with both aCL titer (Rho = 0. 70, P < .0001) and TNF (Rho = 0.84, P < .0001), a measure of disease severity. Excretion of this isoprostane was also higher in those patients who exhibited aPL (P < .0001). Comparable correlations were observed with the isoprostane IPF2alpha -I. No difference of 8-epi-PGF2alpha was observed between patients with and without previous history of thrombosis. This study, showing the existence of a close association between aPL and increased in vivo lipid peroxidation, supports the hypothesis that these antibodies may result from pro oxidative conditions and suggests that inflammation may play an important role. PMID- 9354662 TI - Interacting regions in the A1 and A2 subunits of factor VIIIa identified by zero length cross-linking. AB - Factor VIIIa is a heterotrimer of A1, A2, and A3-C1-C2 subunits, the activity of which is labile due to a weak affinity interaction of the A2 subunit with the A1/A3-C1-C2 dimer. We have used the zero-length cross-linking reagent, 1-ethyl-3 (3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC), to localize regions of interaction within the A1 and A2 subunits. Reaction of factor VIIIa with EDC resulted in the formation of a cross-linked product of approximately 90 kD consisting of the A1 and A2 subunits as judged by Western blotting. Alkaline resistance of this product indicated an amide rather than ester linkage. Factor VIIIa activity decreased as the concentration of cross-linked product increased, suggesting that flexibility in the inter-subunit interaction may be required for proper cofactor function. This product was not formed in the contiguous A1-A2 domains of factor VIII, suggesting that, upon cofactor activation, a conformational change occurs that leads to the formation of a new interdomainal salt bridge(s). Reaction of the EDC-treated factor VIIIa with activated protein C (APC), which cleaves the A1 subunit at Arg336 and bisects the A2 subunit at Arg562, resulted in the formation of an approximately 30 kD product that contains the C-terminus region of A1 covalently linked to the N-terminal half of the A2. The approximately 90 kD cross-linked product was generated after reaction of A2 subunit with A1/A3-C1-C2 dimer but not with A1(336)/A3-C1-C2, a form of the dimer produced by APC cleavage and lacking the C-terminal acidic region of A1. A synthetic peptide corresponding to this acidic region (Met337-Arg372) was found to covalently cross-link to the isolated A2 subunit in 1:1 stoichiometry, suggesting that this region is both necessary and sufficient for the interaction of the A1 and A2 subunits. Sequence analysis of this product suggested that Glu344 in the A1 peptide may contribute to the cross-linkage. These results indicate that activation of factor VIII results in formation of a new ionic linkage(s) localized to the acidic C-terminal region of A1 and the N-terminal half of A2. PMID- 9354661 TI - Regulation of myosin phosphatase through phosphorylation of the myosin-binding subunit in platelet activation. AB - Human platelets were found to contain myosin phosphatase consisting of a 38-kD catalytic subunit of protein phosphatase type 1delta, a 130-kD myosin-binding subunit (MBS) and a 20-kD subunit, all of which cross-reacted with antibodies against these subunits of smooth muscle myosin phosphatase. Anti-MBS antibody coimmunoprecipitated RhoA and Rho-kinase of human platelets. Platelets MBS is a substrate for Rho-kinase and phosphorylation of MBS decreases the activity of myosin phosphatase. Treatment of intact platelets with 9, 11-epithio-11,12 methano-thromboxane A2 led to a dramatic increase in phosphorylation of MBS and a significant decrease in the activity of myosin phosphatase. These findings suggest a putative mechanism for agonist-induced regulation of myosin phosphatase activity in platelets. PMID- 9354663 TI - The human integrin beta3 gene is 63 kb and contains a 5'-UTR sequence regulating expression. AB - The human blood platelet fibrinogen receptor, integrin alphaIIbbeta3 (glycoprotein IIb-IIIa) is an archetypal member of the integrin family of adhesive molecules and is the only integrin encoded by genes physically linked in the genome. Because studies on the normal and abnormal expression of any gene require a thorough understanding of its organization, the initial goals of the current study were to determine the size and complete the genomic organization for the beta3 gene. We now report the isolation of the entire beta3 gene in a single P1 plasmid and for the first time have linked the first and second exons on a contiguous fragment of DNA. Using pulsed-field gel analysis, we determined the full size of the beta3 gene to be 63 kb and show a large (16.7 kb) first intron; based on this information, we propose a uniform numbering system for the beta3 exons. We have completed the 5' genomic structure and generated a long range restriction map. The promoter and the 5' end of the first intron were found to have approximately 50% sequence identity with a region of the avian beta3 gene known to possess functional transcriptional activity. Analysis of three different homologous regions led to the identification of a sequence in the 5'-UTR of the human gene, CCGCGGGAGG, which shares 90% identity with the avian gene and which bound nuclear proteins in DNaseI and electrophoretic mobility shift assay studies. Mutating this sequence caused a 2.6-fold reduction in reporter gene activity. In these studies we have (1) determined the full length and 5' organization of the beta3 gene, (2) identified a large region of homology between the 5' regions of the avian and human genes, and (3) identified a sequence in the 5'-UTR that augments gene expression. Knowing the genomic structure of beta3 has permitted the uncovering of new mechanisms of mutagenesis causing Glanzmann thrombasthenia (Jin et al, J Clin Invest 98:1745, 1996), and our findings will be valuable for such genetic analyses as well as for studies on the transcriptional regulation of beta3 and other integrin genes. PMID- 9354664 TI - A coagulation factor IX-deficient mouse model for human hemophilia B. AB - Coagulation factor IX deficiency causes hemophilia B in humans. We have used gene targeting to develop a coagulation factor IX-deficient (factor IX-knockout) mouse strain. Mouse embryonic stem (ES) cells were targeted by a socket-containing vector that replaces the promoter through exon 3 of the factor IX gene by neoDeltaHPRT, which is a functional neo gene plus a partially deleted hypoxanthine phosphoribosyl transferase minigene. Chimeric mice generated using these socket-containing ES cells transmitted the targeted factor IX gene to their female offspring. Male offspring from these females were characterized and shown to exhibit a phenotype similar to hemophilia B. This factor IX-deficient mouse strain will be useful for studying gene therapy methods and structure-function relationships of recombinant factor IX proteins in vivo. PMID- 9354665 TI - Mutations in the human factor XII gene. AB - The factor XII gene from 31 unrelated factor XII-deficient patients from Germany, Switzerland, and Austria was screened for mutations at the genomic level. Several novel mutations were detected and their absence in a control group of 74 healthy unrelated individuals was checked. Most changes are in the serine protease domain affecting the catalytic triad His-393-Asp-442-Ser-544; two missense mutations, R398Q (arginine 398 to glutamine; gene bank accession no. U71276) and L395M (leucine 395 to methionine; gene bank accession no. U71277), are close to the active site histidine at position 393. Another mutation detected in a cross reacting material (CRM)-positive female with a history of three abortions affects the active site aspartic acid by changing it to asparagine (D442N; gene bank accession no. U71275). The novel mutation G570R (glycine 570 to arginine; gene bank accession no. U71274) giving rise to a CRM-positive phenotype is located next to Cys571, which forms a vital disulfide bridge. Two mutations are causing reading frame shifts: one single basepair deletion in exon 12 [exon 12: 10590(DelC); gene bank accession no. U71278] and one acceptor splice site mutation [exon 14: 11397(G --> A); gene bank accession no. L43615]. The putative regulatory mutation exon 1:-8 (g --> c) in the upstream region of the gene is associated with an aberrant Taq I restriction site allele in intron B of the gene (gene bank accession no. X80393). PMID- 9354666 TI - A recombinant human scFv anti-Rh(D) antibody with multiple valences using a C terminal fragment of C4-binding protein. AB - Monomeric recombinant molecules prove generally unsatisfactory for in vivo use. Most biological systems are indeed multivalent either structurally, associating different chains, or functionally, when cross-linked by their ligands. Mimicking natural molecules for immune intervention implies the need for multimerizing systems to create multivalent molecules capable of interfering with physiological processing. A multivalent anti-Rh(D) recombinant protein has been designed by reconstructing the antibody binding site of a human monoclonal anti-Rh(D) antibody as a single chain Fv mini antibody, then multimerizing it by inserting at its C-terminal end the C-terminal part of the C4 binding protein (C4bp) alpha chain, which is responsible for the octamer multimerization of that molecule. This soluble multivalent recombinant molecule was functional, bound red blood cells (RBCs), agglutinated them, and did not activate complement. This demonstration model opens the way for future in vivo use of multivalent molecules associating antibody valences and other functional molecules for cell targeting, imaging, or removal of cells such as Rh(D)-positive RBCs for preventing Rh alloimmunization. PMID- 9354667 TI - PU.1/Pip and basic helix loop helix zipper transcription factors interact with binding sites in the CD20 promoter to help confer lineage- and stage-specific expression of CD20 in B lymphocytes. AB - CD20 is a B-lineage-specific gene expressed at the pre-B-cell stage of B-cell development that disappears on differentiation to plasma cells. As such, it serves as an excellent paradigm for the study of lineage and developmental stage specific gene expression. Using in vivo footprinting we identified two sites in the promoter at -45 and -160 that were occupied only in CD20+ B cells. The -45 site is an E box that binds basic helix-loop-helix-zipper proteins whereas the 160 site is a composite PU.1 and Pip binding site. Transfection studies with reporter constructs and various expression vectors verified the importance of these sites. The composite PU.1 and Pip site likely accounts for both lineage and stage-specific expression of CD20 whereas the CD20 E box binding proteins enhance overall promoter activity and may link the promoter to a distant enhancer. PMID- 9354668 TI - Structural and functional basis for JAK3-deficient severe combined immunodeficiency. AB - Mutations of the Janus family kinase JAK3 have been found to be responsible for autosomal recessive severe combined immunodeficiency (SCID) in humans. We report here the analysis of four new unrelated patients affected by JAK3-deficient SCID. The genetic defects were heterogeneous and included a large intragenic deletion as well as different point mutations, leading to missense substitutions, early stop codons, or splicing defects. We performed a series of studies of the biochemical events induced by cytokines on lymphoblastoid B-cell lines obtained from these patients. Abnormalities in tyrosine phosphorylation of JAK3 in response to interleukin-2 (IL-2) and IL-4 were present in all patients. Accordingly, IL-2-mediated phosphorylation of STAT5 was also absent or barely detectable. On the contrary, in all cases, we could show reduced but clear phosphorylation of STAT6 upon IL-4 stimulation. In one patient carrying a single amino acid change (Glu481Gly) in the JH3 domain of JAK3, we observed partially conserved IL-2 responses resulting in reduced but detectable levels of JAK3 and STAT5 phosphorylation. Interestingly, the patient bearing this mutation developed a substantial number of circulating CD4(+)/CD45RO+ activated T lymphocytes that were functionally impaired. In two cases, patients' cells expressed JAK3 proteins with mutations in the JH2 pseudo-kinase domain. A single cysteine to arginine substitution (Cys759Arg) in this region resulted in high basal levels of constitutive JAK3 tyrosine phosphorylation unresponsive to either downregulation by serum starvation or cytokine-mediated upregulation. The characterization of the genetic defects and biochemical abnormalities in these JAK3-deficient patients will help define the role of JAK3 in the ontogeny of a competent immune system and may lead to a better understanding of the JAK3 functional domains. PMID- 9354669 TI - Pooled normal human polyspecific IgM contains neutralizing anti-idiotypes to IgG autoantibodies of autoimmune patients and protects from experimental autoimmune disease. AB - Normal human serum contains IgM antibodies that regulate the natural autoantibody activity of IgG in autologous serum. In the present study, we show that pooled normal human IgM (IVIgM) purified from plasma of more than 2,500 healthy donors and processed in a similar fashion to that of therapeutic preparations of pooled normal human IgG (IVIg) suppresses activity of IgG autoantibodies purified from the serum of patients with autoimmune diseases in vitro. The inhibitory effect of IVIgM was greater or equivalent to that of IVIg on a molar basis. We show that IVIgM contains anti-idiotypic antibodies directed against idiotypic determinants of autoantibodies, in particular by showing that Sepharose-bound IVIgM selectively retained F(ab')2 fragments of IgG autoantibodies. The infusion of (Lewis x Brown-Norway) F1 rats with IVIgM protected the animals against experimental autoimmune uveitis induced by immunization with the soluble retinal S antigen, as evidenced by clinical scoring and histopathological analysis. The present findings provide a rationale for considering pooled IgM for immunomodulation of autoimmune disease. PMID- 9354670 TI - Early onset of immunoglobulin heavy chain gene rearrangements in normal human bone marrow CD34+ cells. AB - To characterize early B-cell precursors in humans, we correlated immunoglobulin heavy chain (IgH) gene rearrangement status with the CD34, CD19, and CD10 cell surface markers. Highly purified adult bone marrow (BM) cell fractions were obtained by two successive rounds of flow cytometric cell sorting, and IgH rearrangements were analyzed by polymerase chain reaction (PCR) amplification. Complete VDJH rearrangements were observed in the CD34+ CD19+ fraction, but not in the more immature CD34+ CD19- fraction. About one quarter of these rearrangements had an open reading frame, thus potentially permitting the synthesis of a mu chain. Partial DJH rearrangements were detected in both CD34+ CD19+ and CD34+ CD19- subsets, although they were less abundant in the latter. When triple labeling was used to better characterize the CD34+ CD19- population, DJH rearrangements were found to be present in the CD34(+) CD10+ CD19- fraction, but not in the more primitive CD34+ CD10- CD19-. These results indicate that IgH gene rearrangements occur in CD34+ BM cells and that they initiate in immature progenitors expressing the CD10, but not yet the CD19 surface antigen. Finally, the presence of IgH gene rearrangements in CD34+ BM cells provides a useful marker of clonality to evaluate the possible involvement of these cells in various B-cell lymphoid malignancies. PMID- 9354671 TI - Evidence for copurification of HERV-K-related transcripts and a reverse transcriptase activity in human platelets from patients with essential thrombocythemia. AB - We have previously reported that particles resembling retroviral particles and possessing an RNA-directed DNA polymerase activity can be prepared from platelets. Furthermore, we and others have shown that these particles are present at higher levels in patients with essential thrombocythemia and polycythemia vera. We show here that these particles package RNA molecules that encode HERV-K related pol genes. A subset of the RNA molecules that are packaged are likely to encode the RNA directed DNA polymerase activity and, because these RNAs possess long/full-length open reading frames for the reverse transcriptase and RNaseH (also for part of the integrase domains in genomic clones) of HERV-K, we propose that these transcripts are indeed strong candidates for encoding the enzyme activity found in these particles. Moreover, by using a modification of the polymerase chain reaction-based reverse transcriptase assay in which activated DNA is added during cDNA synthesis to suppress DNA polymerase-mediated RNA directed DNA synthesis, we have found that the particle-associated enzyme behaves like a retroviral reverse transcriptase, further supporting the conclusion that retrovirus-like, perhaps HERV-K sequences, encode this enzyme activity. PMID- 9354672 TI - Markedly reduced expression of platelet c-mpl receptor in essential thrombocythemia. AB - Thrombopoietin (TPO) is implicated as a primary regulator of megakaryopoiesis and thrombopoiesis through binding to the cytokine receptor c-Mpl (the product of the c-mpl proto-oncogene). In an effort to determine the pathophysiological role of TPO-c-Mpl system in essential thrombocythemia (ET), we have examined the levels of serum TPO and the expression and function of platelet c-Mpl in 17 patients with ET. In spite of extreme thrombocytosis, serum TPO levels were slightly elevated or within normal range in most, if not all, patients with ET (mean +/- SD, 1.31 +/- 1.64 fmol/mL), as compared with normal subjects (0.76 +/- 0.21 fmol/mL). Flow cytometric and Western blot analyses revealed that the expression of platelet c-Mpl was strikingly reduced in all patients with ET. Furthermore, the expression of platelet c-mpl mRNA was found to be significantly decreased in the ET patients tested. In contrast, almost identical levels of GPIIb/IIIa protein and mRNA were expressed in platelets from ET patients and normal controls. In addition to expression level, activation state of platelet c-Mpl was investigated in ET patients. Immunoblotting with anti-phosphotyrosine antibody showed that no aberrant protein-tyrosine phosphorylation was observed in platelets of ET patients before treatment with TPO, and the levels of TPO-induced protein-tyrosine phosphorylation, including c-Mpl-tyrosyl phosphorylation, roughly paralleled those of c-Mpl expression, suggesting that c-Mpl-mediated signaling pathway was not constitutively activated in platelets of ET patients. These results suggested that the TPO-c-Mpl system may not be directly linked to pathogenesis of ET, and that gene(s) mutated in ET may be important in regulating the levels of c-mpl gene expression in addition to the growth and differentiation of multipotential hematopoietic stem cells. PMID- 9354673 TI - Elevated serum CD44 level is associated with unfavorable outcome in non-Hodgkin's lymphoma. AB - CD44 molecule is a cell surface glycoprotein involved in many cell-cell and cell matrix interactions. Circulating serum CD44 (s-CD44) levels have been found to change in parallel with response to therapy, but little is known about the predictive or prognostic significance of s-CD44. In the present study, we measured s-CD44 levels in sera taken before treatment from 194 patients with non Hodgkin's lymphoma using a chemiluminescence-enzyme immunoassay method. All except 1 patient were regularly followed-up after therapy at least for 60 months (range, 33 to 143 months). The median pretreatment s-CD44 level was 440 ng/mL (range, 13 to 1,220 ng/mL). Only 32% of the 92 patients with an International Prognostic Index (IPI) score of 0 or 1 had an s-CD44 concentration higher than the median as compared with 67% of the patients with an IPI score >/=2 (P < .0001). Patients with lower than the median s-CD44 achieved more often a complete remission to therapy (P = .0002) and had better survival (P = .007) than those with higher s-CD44 levels. However, in a multivariate analysis, only the IPI score had independent prognostic value (P < .001). The findings were similar if only the patients with diffuse large-cell lymphoma (n = 51) were included in the analysis, but among patients with low-grade lymphoma, the median s-CD44 level was not significantly associated with the IPI or survival. In conclusion, a high s CD44 level at diagnosis is associated with a high IPI score, poor response to treatment, and unfavorable outcome in non-Hodgkin's lymphoma. PMID- 9354674 TI - Immunocytochemical diagnosis of acute promyelocytic leukemia (M3) with the monoclonal antibody PG-M3 (anti-PML). AB - Acute promyelocytic leukemia (APL) is characterized by a reciprocal 15; 17 chromosomal translocation, which fuses the promyelocytic leukemia (PML) and retinoic acid receptor alpha (RARalpha) genes, leading to the expression of the PML/RARalpha fusion oncoprotein. Immunocytochemical labeling of the wild-type PML protein with the PG-M3 monoclonal antibody (MoAb) directed against the amino terminal portion of the human PML gene product, produces a characteristic nuclear speckled pattern that is due to localization of the protein into discrete dots (5 to 20 per nucleus), named PML nuclear bodies. The architecture of PML nuclear bodies appears to be disrupted in APL cells that bear the t(15; 17), thus resulting in a change of the nuclear staining pattern from speckled (wild-type PML protein) to microgranular (PML-RARalpha fusion protein). To assess whether the PG-M3 MoAb could assist in the diagnosis of APL (M3), bone marrow and/or peripheral blood samples from 100 cases of acute nonlymphoid leukemias of different subtypes were blindly immunostained with the PG-M3 MoAb, using the immunoalkaline phosphatase (APAAP) or immunofluorescence technique as detection system. Notably, the abnormal (micropunctate) pattern of the PML/RARalpha fusion protein (usually >/=50 small granules/per nucleus) was observed in APL (M3) samples, but not in other types of acute nonlymphoid leukemias. Immunocytochemical labeling with PG-M3 was particularly useful in the diagnosis of microgranular variant of APL (M3V) (three cases misdiagnosed as M4 and M5), and also to exclude a morphologic misdiagnosis of APL (six of 78 cases). In all cases investigated, immunocytochemical results were in agreement with those of reverse transcription-polymerase chain reaction (RT-PCR) for PML/RARalpha. Because the epitope identified by PG-M3 is located in the aminoterminal portion of PML (AA 37 to 51), the antibody was suitable for recognizing APL cases characterized by breakpoint occurring at different sites of PML (bcr 1, bcr 2 and bcr 3). In conclusion, immunocytochemical labeling with PG-M3 represents a rapid, sensitive, and highly-specific test for the diagnosis of APL that bears the t(15; 17). This should allow an easy and correct diagnosis of this subtype of acute leukemia to any laboratory provided with a minimal equipment for immunocytochemistry work. PMID- 9354675 TI - Folate deficiency delays the onset but increases the incidence of leukemia in Friend virus-infected mice. AB - Clinical studies have indicated that folate deficiency may enhance the development of various malignancies. In animal studies that examined the effect of folate deficiency on malignancies, conflicting results have been reported. In some studies, folate deficiency increased the development and growth of malignant tumors; in others, it decreased the development and growth of malignancies. We examined the effect of transient folate deficiency on the development of leukemia in mice infected with the anemia-inducing strain of Friend leukemia virus. Friend virus disease can be considered as a model for human acute leukemias that are preceded by a preleukemic period. These include leukemias that develop in patients who received previous chemotherapy and/or radiation therapy, as well as patients with chronic granulocytic leukemia or myelodysplasia. Folate deficiency around the time of Friend virus-infection delayed the onset but increased the incidence of leukemia. The rates of rearrangement of the Spi-1 (PU.1 ) oncogene by provirus integration and alteration of the p53 tumor-suppressor gene were the same in leukemia cell lines derived from folate-deficient mice as they were in cell lines from control mice. These results indicate that folate deficiency did not exert its enhancement of leukemogenesis through changes in either Spi-1 or p53, even though these two genes have been found to be the most frequently altered ones in Friend virus-induced leukemias. Our results suggest that folate deficiency may enhance the development of acute leukemia in patients who are at high risk for this disease. PMID- 9354676 TI - A novel chromosomal translocation t(4; 14)(p16.3; q32) in multiple myeloma involves the fibroblast growth-factor receptor 3 gene. AB - Chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus at chromosome 14q32 represent a common mechanism of oncogene activation in lymphoid malignancies. In multiple myeloma (MM), the most consistent chromosomal abnormality is the 14q+ marker, which originates in one third of cases through a t(11; 14)(q13; q32) chromosomal translocation; in the remaining cases, the identity of the partner chromosomes has not been well established. We used a Southern blot approach based on the linkage analysis of the joining (J) and the constant (C) mu, alpha, and gamma regions to detect cases bearing IGH switch mediated chromosomal translocations. We evaluated DNA of 88 nonkaryotyped patients with MM (78 cases) or plasma cell leukemia (PCL) (10 cases) and found the presence of "illegitimate" rearranged IGH fragments (no comigration between the J and C regions) in 21 cases. To confirm this analysis, we cloned the illegitimate rearranged fragments from three samples, and the molecular and fluorescent in situ hybridization (FISH) analyses indicated the presence of chromosomal translocations juxtaposing a switch IGH region to sequences from chromosomes 11q13 (one PCL case) or 4p16.3 (two MM cases). Interestingly, the breakpoints on 4p16.3 occurred about 14 kb apart in a genomic region located approximately 50 kb centromeric to the fibroblast growth-factor receptor 3 (FGFR3) gene. Moreover, Southern blot analysis using 4p16.3 genomic probes detected a rearrangement in an additional MM tumor. FISH analysis of the MM derived KMS-11 cell line, reported to be associated with a t(4; 14)(p16.3; q32), showed that the FGFR3 gene was translocated on 14q32. High levels of FGFR3 mRNA expression were observed in the cloned MM tumors and KMS-11 cell line, but not in the cases that were apparently negative for this lesion. Furthermore, a point mutation at codon 373 in the transmembrane domain of the FGFR3 gene resulting in an amino acid substitution (Tyr --> Cys) was detected in the KMS-11 cell line. These findings indicate that the t(4; 14)(p16.3; q32) represents a novel, recurrent chromosomal translocation in MM, and suggest that the FGFR3 gene may be the target of this abnormality and thus contribute to tumorigenesis in MM. PMID- 9354677 TI - Preferential dissemination of B-cell gastric mucosa-associated lymphoid tissue (MALT) lymphoma to the splenic marginal zone. AB - The tendency for gastric mucosa-associated lymphoid tissue (MALT) lymphoma cells preferentially to localize around reactive B-cell follicles, both in the mucosa and regional lymph nodes, coupled with their immunophenotype, has led to the proposal that the normal cell counterpart of this lymphoma is the marginal zone B cell. In keeping with this proposition, lymphocytes expressing the lymphoma idiotype have been detected in the splenic marginal zone in a single case of gastric MALT lymphoma. To confirm that this truly represented preferential homing of MALT lymphoma to the splenic marginal zone, we have now re-examined this case, together with 17 other cases, using both immunohistochemical and molecular methods in an attempt to establish clonal identity between the gastric lymphoma and cells in the splenic marginal zone. In three cases, the spleen was characterized by marked expansion of marginal zones by cells showing the same pattern of Ig light chain restriction as the gastric lymphoma. None of the remaining 15 cases showed histologic evidence of lymphomatous infiltration. Analysis of the Ig genes by polymerase chain reaction (PCR), cloning, and sequencing confirmed clonal identity between the splenic marginal zone infiltrates and the gastric lymphoma in the histologically involved cases. Amplifiable DNA could be extracted from only 5 of the remaining 15 cases. In 3 of these cases, including the case previously studied using an anti-idiotype, involvement of the splenic marginal zone could be confirmed using microdissection and clone-specific PCR. No involvement could be detected in the remaining 2 cases. In addition, we have shown that mucosal addressin cell adhesion molecule-1 (MAdCAM-1), the primary homing receptor of gut-mucosa for lymphocytes, was strongly expressed by the sinus lining cells of the splenic marginal zone. These results provide strong evidence for preferential involvement of the marginal zone when gastric MALT lymphomas disseminate to the spleen, which is in keeping with the notion that the marginal zone B cells are the normal counterparts of MALT lymphoma cells. PMID- 9354678 TI - Correlation between mutation in P53, p53 expression, cytogenetics, histologic type, and survival in patients with B-cell non-Hodgkin's lymphoma. AB - In the biology of a cell, the central role of p53 in controlling functions such as G1/S transition (check point) and DNA damage repair, and as a trigger of apoptosis, is well established. Somatic mutations or other changes in P53 have been reported in numerous tumor types, and in some of these, they are associated with poor prognosis. In this study, we examined 237 cytogenetically characterized B-cell non-Hodgkin's lymphomas (B-NHLs) for somatic changes in P53 by Southern blot analysis, by single-strand conformation polymorphism analysis (SSCP) of exon 5 through 9, and by direct sequencing of SSCP variants to determine the frequency and types of mutations and their clinical significance. In a portion of these (173 tumors), we also studied p53 expression by immunostaining. On Southern blots, no gross change was identified in P53 and no mutation was identified in exon 9. In exons 5 through 8, 27 different mutations were identified in 25 patients (23 single-base substitutions, 3 deletions, 1 duplication). Mutations in P53 were identified in 25 of 237 tumors (10.5%), which included 1 of 45 small lymphocytic lymphomas (SLLs), 2 of 38 follicular small cleaved-cell lymphomas (FSCCs), 2 of 35 follicular mixed small cleaved-cell and large-cell lymphomas (FMxs), 1 of 4 follicular large-cell lymphomas (FLCs), 1 of 14 diffuse small cleaved-cell lymphomas (DSCCs), 2 of 17 diffuse mixed small- and large-cell lymphomas (DMxs), and 16 of 84 diffuse large-cell lymphomas (DLCCs); the difference between the histologic groups was significant (P < .01). Among mantle cell lymphoma (MC) patients, 3 of 10 had mutations. In 16 patients, the mutation was identified in specimens obtained at diagnosis. Mutation of transition type and transversion type occurred at a relative frequency of 2:1. Thirty percent occurred at CpG dinucleotide sequences and the codon for arginine was most frequently affected. Nineteen of 99 tumors with complex cytogenetic abnormalities, but none of 69 tumors with simple cytogenetic abnormalities, had mutations (P < .001). Similarly, 11 of 25 tumors with an abnormality of 17p and 8 of 143 tumors with apparently normal 17p had mutations (P < .0001). Positive correlations were found between a mutation and p53 expression (P < .001), between missense type mutations and p53 expression (P < .005), and between 17p abnormalities and p53 expression (P < .05). Twenty-two of 49 patients without mutation and 14 of 17 patients with mutations died (P < .05), but there was no significant difference in median survival. Similarly, 21 of 26 p53 positive patients died, whereas only 1 of 24 p53-negative patients died on-study (P < .001). Among p53-negative patients, mutation (P < .01) was positively associated with a fatal outcome. These findings indicate that in B-NHL, somatic changes in P53 were present in diagnostic specimens of all histologic types, but at a higher frequency in DLC and MC tumors. P53 mutation and/or expression has a negative influence on survival, and therefore can serve as prognostic indicators. Immunostaining for p53 is an effective way to screen for P53 changes in these tumors. PMID- 9354679 TI - The plasmacytoma resistance gene, Pctr2, delays the onset of tumorigenesis and resides in the telomeric region of chromosome 4. AB - Mouse plasmacytomas share pathogenetic features in common with both multiple myeloma and Burkitt's lymphoma in humans. Susceptibility to plasmacytoma induction by intraperitoneal pristane in mice is controlled by multiple genes. At least two of these genes reside on mouse chromosome 4 in regions of the genome sharing linkage homology with human chromosomes 9p21, 1p32, and 1p36. A series of congenic strains recombinant for regions of mouse chromosome 4 in the vicinity of the Pctr2 predisposition locus were created and typed for their tumor susceptibility/resistance phenotypes. These strains were derived by introgressively backcrossing alleles from resistant DBA/2 mice onto the susceptible BALB/cAnPt background. Six resistant and two susceptible strains were allelotyped for 10 genes and 49 random DNA markers to identify the smallest region of overlap in the resistant strains. These studies have determined that the Pctr2 locus resides in either a 500-kb interval proximal to Nppa, or in a 1- to 2-centiMorgan (cM) interval distal to Nppa. In these congenic strain analyses, the Nppa and Fv1 loci, in addition to genes within about 1 cM of these loci, have been excluded as candidates for the Pctr2 locus. A relevant locus that may reside in this interval is Rep2; it is associated with the efficiency of repairing X-ray induced DNA damage sustained during the G2 phase of the mitotic cycle. The Pctr2 locus acts in a codominant fashion. F1 hybrids between resistant and susceptible congenic strains exhibit a reduced tumor incidence and a significant delay in the onset of tumorigenesis. Identification and eventual cloning of the Pctr2 locus may assist in the identification of genes involved in many types of cancer showing aberrations in human chromosome 1p36. PMID- 9354681 TI - p16INK4A promotes differentiation and inhibits apoptosis of JKB acute lymphoblastic leukemia cells. AB - Homozygous p16(INK4A) (p16) gene deletion is frequent in primary tumor cells from acute lymphoblastic leukemia (ALL), suggesting that loss of p16 may be an important precursor to transformation in ALL. We have previously described JKB, a human ALL cell line, that contains homozygous deletion of the p16 gene. Because ectopic expression of p16 suppresses cell growth, we created a temperature sensitive p16 mutant to develop a system for inducible p16 function in human ALL. JKB cells were transfected either with a p16 gene mutated at position 119 (E119G) to confer temperature sensitivity (JKB p16MT) or with control vector. The percentage of cells in G1 phase was similar in JKB control cells or in JKB p16MT cells cultured at restrictive conditions (40 degrees C). However, with lowering of temperature from 40 degrees C to permissive conditions (31 degrees C), the percentage of JKB p16MT cells in G1 phase and binding of p16 to CDK4 and CDK6 increased, with associated decreases in CDK4 and CDK6 kinase activities, and dephosphorylation of retinoblastoma protein (pRB). Culture of JKB p16MT cells at 31 degrees C for >/=3 days irreversibly inhibited growth. Moreover, JKB p16MT cells cultured under these permissive conditions showed a less transformed morphology and more differentiated phenotype than did these cells cultured under restrictive temperatures. Finally, dexamethasone (Dex) induced apoptosis of JKB p16MT cells cultured at 40 degrees C, but did not trigger death of these cells cultured at 31 degrees C. These results suggest that deletion of p16 gene in JKB human ALL cells is associated with dysregulated growth of less differentiated tumor cells, which nonetheless remain susceptible to apoptosis triggered by Dex. PMID- 9354680 TI - The role of Mig, the monokine induced by interferon-gamma, and IP-10, the interferon-gamma-inducible protein-10, in tissue necrosis and vascular damage associated with Epstein-Barr virus-positive lymphoproliferative disease. AB - The mechanisms of tissue necrosis and vascular damage characteristics of certain Epstein-Barr virus (EBV)-associated lymphoproliferative disorders are unknown. The CXC chemokines interferon-gamma-inducible protein-10 (IP-10) and the monokine induced by interferon-gamma (Mig) caused tissue necrosis and vascular damage in Burkitt's lymphoma tumors established in nude mice. We report higher levels of IP 10 and Mig gene expression in tissues with necrosis and vascular damage from EBV positive lymphomatoid granulomatosis and nasal or nasal-type T/natural killer (NK)-cell lymphomas compared with tissues with lymphoid hyperplasia, which lacked tissue necrosis and vascular damage. By immunohistochemistry, Mig and IP-10 proteins localized with similar patterns in viable tissue surrounding dead tissue, mostly within endothelial cells, monocyte/macrophages, and lymphocytes. Circulating levels of IP-10 were abnormally elevated in patients with EBV positive lymphomatoid granulomatosis and nasal or nasal-type T/NK-cell lymphomas. These experiments provide the first description of the presence of Mig in any human normal or diseased tissue and the first description of IP-10 in certain lymphoproliferative lesions. These data suggest that Mig and IP-10 play an important role in the pathogenesis of tissue necrosis and vascular damage associated with certain EBV-positive lymphoproliferative processes. PMID- 9354682 TI - HOXC4, HOXC5, and HOXC6 expression in non-Hodgkin's lymphoma: preferential expression of the HOXC5 gene in primary cutaneous anaplastic T-cell and oro gastrointestinal tract mucosa-associated B-cell lymphomas. AB - Most of the 39 members of the homeobox (HOX) gene family are believed to control blood cell development. HOXC4 and HOXC6 gene expression levels increase with differentiation of lymphoid cells. In contrast, HOXC5 is not expressed in the lymphoid lineage, but was found in lymphoid cell lines, representing the neoplastic equivalents of various differentiation stages of T and B lymphocytes. In the present study, we investigated the HOXC4, HOXC5, and HOXC6 gene expression pattern in 89 non-Hodgkin's lymphomas (NHLs) of different histologic subtypes and originating from different sites. Using RNA in situ hybridization and semiquantitative reverse transcription-polymerase chain reaction, we found expression of HOXC4 in 83 of 88 and HOXC6 in 77 of 88 NHLs and leukemias investigated. In contrast, HOXC5 expression was found in only 26 of 87 NHLs and appeared to be preferentially expressed by two specific subsets of lymphomas, ie, primary cutaneous anaplastic T-cell lymphomas (9 of 9) and extranodal marginal zone B-cell lymphomas (maltomas; 7 of 9). These results indicate that, in contrast to HOXC4 and HOXC6, HOXC5 shows a type- and site-restricted expression pattern in both T- and B-cell NHLs. PMID- 9354683 TI - Biochemical and molecular characterization of hereditary myeloperoxidase deficiency. AB - Hereditary myeloperoxidase (MPO) deficiency is a neutrophil disorder characterized by the lack of peroxidase activity. Cytochemical, biochemical, spectroscopic, immunocytochemical, and genetic studies were carried out on a 5 year-old MPO-deficient subject and on her parents. The father was also MPO deficient, whereas the mother had 24% of normal MPO activity. Although the typical absorption spectrum of MPO was absent in both the father and daughter, the father's neutrophils, and not those of the daughter, contained material antigenically related to MPO. In the MPO gene of the father, two mutations were found, each located in a different allele: a T --> C transition, causing the nonconservative replacement M251T and a 14-base deletion within exon 9. The M251T substitution occurred in the carboxy-terminal region of the light chain that is included in the heme pocket. The daughter inherited the 14-base deletion from her father. The study of the MPO mRNAs present in liquid cultures of granulocyte precursors surprisingly showed that the same genetic defect, ie, the 14-base deletion, seemed to exhibit different mRNA phenotypes in the father and the daughter. In fact, mRNA derived from the 14-base-deleted allele was not found in the father and an aberrantly spliced MPO mRNA with a 77-base deletion of exon 9, which includes the 14-base deletion and leads to the generation of a premature stop codon, was found in the daughter. The possibility that Delta77 mRNA could derive from other mutations linked to the Delta14 allele was dismissed because no sequence differences were found in the region (exons and exon-intron junctions). Our data indicate that the alteration of the mRNA context caused by the 14-base deletion provide a basis for the 77-base deletion in the mRNA processing. Since the granulocyte precursors from the liquid cultures of the father were more differentiated than those from the daughter, the observed different behavior of the 14-base-deleted allele in the father and daughter may be the result of a differentiation-stage dependent control of altered spliced mRNA, which may be tolerated during the early stages of differentiation but degraded at later stages. In the liquid cultures of the daughter's cells, in addition to the mRNA with the 77-base deletion, a mRNA with the wild type sequence was also found. This mRNA was inherited from the mother, since no mutations were found in her MPO cDNA and MPO gene. The MPO defect might be caused by a regulatory mutation that induces the MPO gene switch off at an early stage of granulocyte differentiation. PMID- 9354684 TI - Murine macrophage mannose receptor promoter is regulated by the transcription factors PU.1 and SP1. AB - The mannose receptor (MR) is a transmembrane protein that functions primarily as a phagocytic receptor for a wide range of microorganisms. Its expression appears to be restricted to tissue macrophages and Langerhans cells. To gain an understanding of the regulation of the gene, we have isolated the 5' flanking sequence of the murine MR gene and have analyzed a 536-bp sequence upstream of the ATG start site for transcriptional activity. This sequence lacks a TATA box but contains an initiator (Inr) consensus element overlapping the single transcriptional start site. Transcription factor binding sites contained within this sequence include PU.1, Sp1, ETS, GATA, and MYB motifs. Serial 100-bp deletions of this promoter fragment fused to a luciferase reporter gene showed various patterns of activity when transfected into different cell types. In myeloid cells, sequence elements upstream of bp -300 appeared to have a silencing effect on promoter activity. Of the four potential PU.1 binding sites contained within the fragment, one site (at -164) bound the PU.1 factor most strongly, whereas the adjacent PU.1 site (at -177 bp) bound PU.1 to a lesser degree. Mutations of these sites decreased transcriptional activity but did not abolish it. However, promoter activity was abrogated when both the -164 bp PU.1 site and the adjacent -177 bp PU.1 site were mutated. In addition, mutation of the Sp1 site also significantly reduced promoter activity. Cotransfection studies in Drosophila Schneider cells indicated that PU.1 and Sp1 may function synergistically in transactivating the murine MR. This study indicates that MR gene expression is regulated in part by the interaction between the ubiquitously expressed factor Sp1 and the lymphoid/myeloid factor PU.1 and provides a basis for studying the regulation of this gene. PMID- 9354686 TI - Intracellular pool of vascular endothelial growth factor in human neutrophils. AB - Vascular endothelial growth factor (VEGF ), an endothelial cell mitogen, is a potent angiogenic factor produced by several cell types. Whether human neutrophils are potential producers of VEGF has not yet been described. The present work shows that phorbol-12-myristate 13-acetate (PMA), fMet-Leu-Phe, and tumor necrosis factor-alpha (TNF-alpha) triggered a time-dependent secretion of VEGF by human neutrophils. Cells incubated with 50 ng/mL of PMA released significant amounts of VEGF after 15 minutes. Because the extracellular content of VEGF in human neutrophils supernatants remained constant over a period of 2 to 24 hours and because PMA is a potent inducer of human neutrophil degranulation, the PMA-induced secretion of VEGF may be due to a pre-existing intracellular pool of this molecule. This hypothesis was reinforced by the absence of cycloheximide effect on the PMA-induced secretion of VEGF. The existence of an intracellular pool of VEGF was confirmed by measuring the intracellular content of VEGF in resting neutrophils. A dosedependent inhibition of PMA-induced VEGF secretion was observed when the cells were incubated in the presence of pentoxifylline, a methylxanthine known to inhibit neutrophil degranulation. To confirm the implication of neutrophil degranulation in VEGF release, the effects of two inducers of physiologic degranulation, fMet-Leu-Phe and TNF-alpha, were determined. Both agonists induced a release of VEGF in the absence of cytochalasin B, confirming the involvement of neutrophil degranulation and suggesting the intracellular localization of VEGF in the specific granule fraction. In addition, the kinetics of fMet-Leu-Phe- and TNF-alpha-induced secretion of lactoferrin were similar to those of VEGF release induced by these two both agonists. The subcellular fractionation of human neutrophils showed a granule-specific distribution of the intracellular pool of VEGF in resting neutrophils. The finding that human neutrophils contain an intracellular pool of VEGF, secreted in the extracellular space under PMA-, fMet-Leu-Phe-, and TNF alpha-induced degranulation, suggests a role for human neutrophils as cellular effectors of physiologic as well as pathologic angiogenesis. PMID- 9354685 TI - Tumor necrosis factor alpha-induced eosinophil accumulation in rat skin is dependent on alpha4 integrin/vascular cell adhesion molecule-1 adhesion pathways. AB - Tumor necrosis factor alpha (TNFalpha) is a cytokine implicated in the pathogenesis of numerous chronic and acute inflammatory conditions. In the present study, we have characterized the ability of TNFalpha in inducing eosinophil accumulation in rat skin and have shown the inhibitory effects of anti alpha4 integrin and anti-vascular cell adhesion molecule-1 (VCAM-1) antibodies on this response. The intradermal injection of recombinant human TNFalpha induced a slowly developing, dose-dependent accumulation of 111In-eosinophils in rat skin that was maximal at the dose of 10(-11) mol/site. Coadministration of TNFalpha with the soluble TNFalpha receptor (p55)-IgG fusion protein (TNFR-IgG) totally inhibited the 111In-eosinophil accumulation induced by the cytokine. The TNFalpha induced 111In-eosinophil accumulation was not affected after pretreatment of rats with the platelet-activating factor (PAF) receptor antagonist UK-74,505 or the antihuman interleukin-8 monoclonal antibody (MoAb) DM/C7. In contrast, the intravenous administration of an anti-alpha4 integrin MoAb, HP2/1 (3.5 mg/kg), or an anti-VCAM-1 MoAb, 5F10 (2 mg/kg), greatly inhibited the 111In-eosinophil accumulation induced by TNFalpha (the responses detected at 10(-11) mol/site were inhibited by 78% and 50%, respectively). These results show that TNFalpha is an effective inducer of eosinophil accumulation in vivo, with this response being dependent on an interaction between alpha4 integrins and VCAM-1. PMID- 9354687 TI - Interleukin-10 upregulates tumor necrosis factor receptor type-II (p75) gene expression in endotoxin-stimulated human monocytes. AB - Interferon-gamma (IFN-gamma) upregulates expression of certain genes in monocytes, including cell-surface molecules such as HLA class II, B7, and ICAM-1. IFN-gamma also potentiates production of cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta) and IL-12. Conversely, IL-10 downregulates expression of many of these same genes and often antagonizes the effects of IFN-gamma. IL-10 is known to inhibit TNF-alpha production in lipopolysaccharide (LPS)-stimulated monocytes; however, the effects of IL-10 on TNF receptor (TNF-R) expression are not well defined. We examined the effects of IL-10 on production of both membrane-associated (m) and soluble (s) TNF-R type II (sTNF-RII) by purified human CD14(+) monocytes. We also compared the effects of IFN-gamma and IL-10 on production of TNF-alpha and sTNF-RII by these cells. Monocytes constitutively expressed low levels of TNF-RII mRNA and mTNF-RII protein. LPS stimulation induced rapid, but transient loss (shedding) of mTNF-RII molecules and a delayed, but marked increase in TNF-RII mRNA levels. IL-10 increased expression of both mTNF-RII and sTNF-RII by LPS-stimulated monocytes, whereas IFN-gamma decreased their expression. The increased levels of sTNF-RII in cultures of IL-10-treated monocytes correlated directly with increased levels of TNF-RII mRNA and inversely with the levels of TNF-alpha mRNA. The ability of IL 10 to upregulate TNF-RII gene expression was transcriptionally mediated because actinomycin D blocked this effect. Furthermore, IL-10 treatment did not alter the half-life of TNF-RII mRNA transcripts in LPS-stimulated monocytes. To further examine the mechanism by which IL-10 potentiates TNF-RII gene expression, a 1.8 kb fragment of the human TNF-RII promoter cloned into a luciferase expression vector (pGL2-basic) was transfected into the IL-10-responsive macrophage cell line, RAW264.7. Although IL-10 alone induced only minimal promoter activity in these cells, it markedly increased the LPS-induced response, providing further evidence that the ability of IL-10 to amplify TNF-RII gene expression is transcriptionally controlled. Together, these findings demonstrate that IL-10 coordinately downregulates expression of TNF-alpha and upregulates expression of TNF-RII, particularly the soluble form of this receptor, in monocytes. PMID- 9354688 TI - Band 3 peptides block the adherence of sickle cells to endothelial cells in vitro. AB - Malaria-parasitized erythrocytes have increased endothelial adherence due to exposure of previously buried intramembranous sites of band 3. Because sickle erythrocytes also show increased adhesiveness and because the membrane portion of band 3 is aggregated in both types of cells, we examined the role of band 3 in sickle cell adhesiveness. Synthetic peptides derived from the second and third exofacial, interhelical regions of band 3 completely inhibited the abnormal adherence of sickle cells to an endothelial monolayer in a static assay. This effect was observed independently of plasma factors, required micromolar levels of peptide, was sequence-specific, and was found with both L- and D-isomers. The active peptides also inhibited the increased adherence induced by low-dose calcium loading of normal red blood cells. Finally, a monoclonal antibody against an active peptide specifically immunostained a fraction of sickle cells. These findings implicate a role for band 3 in at least one type of sickle cell adhesiveness via the exposure of normally cryptic membrane sites. PMID- 9354689 TI - IRC011, a new synthetic chelator with selective interaction with catabolic red blood cell iron: evaluation in hypertransfused rats with hepatocellular and reticuloendothelial radioiron probes and in iron-loaded rat heart cells in culture. AB - A major consideration in the selection of new and improved iron chelators for clinical use is preferential interaction with the most toxic iron compartment. We describe the biologic properties of a new synthetic hexadentate iron chelator (IRC011) that is a substituted polyaza compound. Unlike deferoxamine (DF ), the polyaza structure of IRC011 does not contain any readily hydrolyzable covalent bonds and is anticipated to resist in vivo biotransformation. In the present studies, the ability of IRC011 to remove radioiron from iron-loaded heart cells in vitro was similar to DF, with a decrease to 20.0 +/- 0.4% and 19.7 +/- 0.5% of initial values after 24 hours of incubation with 0.3 mmol/L of DF or IRC011, respectively. The in vivo interaction of IRC011 with specific iron stores was studied in hypertransfused rats using selective labeling of reticuloendothelial (RE) iron stores with 59Fe-heat-denatured red blood cells (DRBCs) and of hepatocellular stores with 59Fe-ferritin. The pattern of radioiron excretion with IRC011 was quite different from that with DF. Although with both compounds, hepatocellular iron excretion was through the bile, whereas RE iron excretion was mainly in the urine, the magnitude of these effects was quite different. After the administration of a single parenteral dose of 200 mg/kg representing a 53% higher iron-binding capacity for IRC011 compared with DF, 48-hour urinary excretion of RE iron with IRC011 was 22.8% +/- 1.1% (% of total body 59Fe), but only 6.0% +/- 3.6% with DF. By contrast, the corresponding biliary excretion of hepatocellular radioiron was 14.2% +/- 3.2% with DF, but only 0.7% +/- 0.3% with IRC011. Thus, the new iron chelator IRC011 is distinguished from DF by the following features: (1) a higher affinity to Fe(III), (2) anticipated resistance to in vivo catabolism, (3) preferential interaction with RE iron derived from RBC breakdown, and (4) selective renal excretion. Because RBC breakdown is the most likely source of the toxic nontranferrin plasma iron, IRC011 may be a useful iron chelator for protecting vital organs from peroxidative damage. PMID- 9354690 TI - The exon 46-encoded sequence is essential for stability of human erythroid alpha spectrin and heterodimer formation. AB - Human erythroid alpha-spectrin alleles responsible for hereditary elliptocytosis (alphaHE alleles) undergo increased incorporation into red blood cell membranes when the polymorphism alphaLELY (LELY: Low Expression LYon) occurs in trans. The alphaLELY polymorphism is characterized by a mutation in exon 40 at codon 1857 (CTA --> GTA, Leu --> Val) and the partial (50%) skipping of exon 46, which encodes residues 2177-2182 (Wilmotte et al, J Clin Invest 91:2091, 1993). Both of these peptide sequence alterations are located within the region of the alpha chain involved in initiating heterodimer assembly, and either or both mutations could potentially contribute to decreased incorporation of alpha-chains from the alphaLELY allele in heterozygotes into red blood cell membranes. These possibilities were evaluated by testing the protease resistance and in vitro binding properties of normal and mutant recombinant 4-motif alpha subunit peptides containing the dimer initiation region. The two forms of alpha spectrin produced by alternative mRNA splicing of the alphaLELY allele were represented by alpha18-21(1857), a peptide with the codon 1857 mutation and retaining the exon 46 encoded sequence, and alpha18-21(1857-Delta46), a peptide carrying both the 1857 codon mutation and the exon 46 deletion. The properties of these two recombinant peptides were compared with alpha18-21, a peptide with the normal sequence at codon 1857 and retaining the exon 46 encoded sequence. The codon 1857 mutation does not adversely affect dimer formation, but it is responsible for the increased trypsin cleavage between the alphaIV and alphaV domains that was the characteristic feature initially used to identify the alphaLELY (SpalphaV/41) polymorphism (Alloisio et al, J Clin Invest 87:2169, 1991). Deletion of the six amino acids encoded by exon 46 perturbs folding of the alpha21 motif, because this region of the alpha18-21(1857-Delta46) peptide is rapidly degraded and this recombinant peptide is unusually prone to self-aggregation. Exon 46 deletion reduces, but does not eliminate, dimerization. Comparison of mild trypsin proteolytic products from an alphaLELY homozygote and the two alphaLELY recombinant peptides strongly suggests that little, if any, of the 50% of the alpha chains from the alphaLELY allele that contain the exon 46 deletion are incorporated into the mature erythroid membrane. Based on the in vitro analysis of recombinant alphaLELY peptides, the inability of detectable amounts of exon 46(-) alpha chains to assemble into the mature membrane skeleton in vivo is probably due to a combination of decreased dimer binding affinity and increased proteolytic degradation of these mutant chains. PMID- 9354691 TI - Exclusion of three candidate genes as determinants of congenital dyserythropoietic anemia type II (CDA-II). AB - Congenital dyserythropoietic anemia type II (CDA-II) is the most common form of inherited dyserythropoiesis. Previous studies have shown that the anion transporter (band 3) is narrower and it migrates faster on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE); this aspect was related to insufficient glycosylation. Biochemical data support the hypothesis that this disease is due to a deficiency of N-acetylglucosaminyltransferase II (GnT II) or alpha-Mannosidase II (alpha-Man II), which represent the key to glycosylation. In addition, a third candidate gene is alpha-Man IIx, which shows a strong homology with alpha-Man II. The knowledge of the chromosomal localization of these putative genes allowed us to perform a linkage study using three sets of microsatellite markers flanking the candidate genes. Six families with two or more affected children were enrolled in this study. The data obtained exclude linkage to all three candidate genes. In consideration of the biochemical data (reduction of enzymatic activity) of the same enzymes, our results suggest the hypothesis that a defect in an unknown transcriptional factor is involved in CDA II. PMID- 9354692 TI - Allogeneic versus autologous bone marrow transplantation for refractory and recurrent low-grade non-Hodgkin's lymphoma. AB - Patients with recurrent or refractory low-grade non-Hodgkin's lymphoma (NHL) are increasingly treated with myeloablative therapy and autologous stem cell transplantation. However, allogeneic bone marrow transplantation (BMT) is only sporadically performed in such patients. Therefore, we wish to compare treatment results of patients with recurrent or refractory low-grade NHL who underwent allogeneic BMT with those who underwent autologous BMT in our center. Twenty eight patients were studied. The patients had received 2 to 5 lines of conventional chemotherapy before the BMT procedure. Eighteen patients, all with chemotherapy-sensitive disease at the time of transplantation, underwent autologous BMT and 10 patients, of whom 7 with chemotherapy-resistant disease at the time of transplantation, underwent allogeneic BMT. Furthermore, all allogeneic BMT patients had overt lymphoma infiltration of the BM at the time of transplantation. The conditioning regimen consisted of cyclophosphamide plus total body irradiation in all 28 patients. All allogeneic BMT patients achieved complete remission, 3 patients had a treatment-related death, and 7 patients are alive and disease-free with a median follow-up of 41 months. In contrast, none of the autologous BMT patients died of transplant-related complications. However, despite the fact that all autologous BMT patients had chemotherapy-sensitive disease and partial remission was converted to complete remission by the BMT procedure in 67% of them, only 3 of 18 patients are alive and disease-free. The probability of relapse or disease-progression among allogeneic BMT patients was 0% compared with 83% for autologous BMT patients (P = .002). Progression-free survival rates 2 years after BMT were 68% for allogeneic BMT patients and 22% for autologous BMT patients (P = .049). Although the numbers of patients are small, this study suggests that allogeneic BMT offers a better chance for cure than autologous BMT for patients with poor-prognosis low-grade lymphoma, and the difference in relapse or disease progression is strongly suggestive for the existence of a graft-versus-low-grade lymphoma effect. PMID- 9354693 TI - Donor leukocyte infusions are effective in relapsed multiple myeloma after allogeneic bone marrow transplantation. AB - Donor leukocyte infusions (DLI) can induce sustained remissions in patients with acute and chronic myeloid leukemia who relapse after allogeneic bone marrow transplantation (allo-BMT). Also, in multiple myeloma (MM), incidental reports have indicated the existence of a graft-versus-myeloma effect (GVM) induced by allo-reactive T cells. We performed a retrospective study in a larger group of MM patients to characterize better the effect, prognostic factors, and toxicity of this new treatment modality. Thirteen patients with relapsed MM after allo-BMT were studied. Patients received a total of 29 DLI with T-cell doses ranging from 1 x 10(6)/kg to 33 x 10(7)/kg. Repetitive courses, sometimes with escalated cell doses, were undertaken in case of no response to or relapse after DLI. Eight of 13 patients responded: 4 patients achieved a partial remission and 4 patients achieved a complete remission. Dose escalation was effective in 3 patients. The time to response was median 6 weeks (range, 4 to 10 weeks). Major toxicities were secondary to acute and chronic graft-versus-host disease (GVHD), which occurred in 66% and 56% of all patients and in 87% and 85% of the responders, respectively. Two responding patients developed fatal BM aplasia. The only prognostic factors for response were a T-cell dose greater than 1 x 10(8)/kg and the occurrence of GVHD. Seven of nine patients developing acute GVHD responded, as compared with only 1 response in the 4 patients without GVHD and 6 of 7 patients with chronic GVHD responded, whereas no response was observed in the 5 patients without chronic GVHD. DLI are effective in a high percentage of patients with relapsed MM after allo-BMT, although it is associated with a high treatment related toxicity. The dose of T cells used may be important in determining the GVM effect, with the highest probability of response after infusion of more than 1 x 10(8) T cells. Because the optimal individual dose may vary, patient-adapted therapy consisting of repeated infusions with escalating dose of donor leukocytes until maximum response is achieved may therefore be preferable. PMID- 9354694 TI - Failure of immunologic purging in mantle cell lymphoma assessed by polymerase chain reaction detection of minimal residual disease. AB - To assess the clinical significance of minimal residual disease (MRD) detection by polymerase chain reaction (PCR) we analyzed samples from 26 patients with mantle cell lymphoma (MCL) who had undergone bone marrow transplantation (BMT) at the Dana-Farber Cancer Institute. The BCL-1/IgH translocation and clonally rearranged Ig heavy chain genes (IgH) provided molecular markers for detection and follow-up of MRD by polymerase chain reaction (PCR) amplification in 19 of the 26 (73%) MCL patients studied. IgH gene sequencing analysis showed somatic mutations in MCL that are characteristic of an antigen driven process suggesting that, in MCL, the final malignant transformation occurs in a mature B cell. Of the 19 patients with a PCR amplifiable marker, 17 underwent autologous, 1 an allogeneic, and 1 a syngeneic bone marrow transplantation (BMT). All patients had PCR-detectable MRD in the bone marrow (BM) at the time of BMT, irrespective of any history of histological BM involvement. In contrast to other B-cell malignancies, we found that immunological purging with complement-mediated lysis eradicated PCR-detectable MCL in only two patients. Moreover reinfusion of MRD was associated with a poor outcome. More than half of the patients undergoing autologous BMT had relapsed by the time of restaging at 2 years after autologous BMT. In four MCL patients in whom no residual lymphoma was reinfused, including the allogeneic and the syngeneic BMT, only one patient relapsed. Persistence of MRD detection after BMT was also associated with a high probability of relapse, although one patient did not have PCR-detectable MRD in peripheral blood or BM before relapse at nodal sites. We conclude that PCR amplification of disease specific markers is a feasible and sensitive method to assess MRD and its clinical significance in patients with MCL. Moreover, PCR amplification provides a tool to evaluate modifications of purging and stem cell collection procedures that may be required for the management of this otherwise incurable disease. PMID- 9354695 TI - Altered tyrosine phosphorylation via the very late antigen (VLA)/beta1 integrin stimulation is associated with impaired T-cell signaling through VLA-4 after allogeneic bone marrow transplantation. AB - Our previous study showed that the cross-linking of very late antigen (VLA)/beta1 with anti-CD29 monoclonal antibody (MoAb), or interactions with extracellular matrix (ECM) proteins through VLA/beta1, failed to induce T-cell costimulation via the CD3/T cell receptor (TCR) pathway for over 1 year after allogeneic bone marrow transplantation (allo-BMT), although normal CD29 and CD3 expression was observed after 3 months following allo-BMT. Molecular analysis revealed altered tyrosine phosphorylation of cellular proteins by the solid-phase cross-linking of VLA/beta1 molecules in T cells from patients after allo-BMT. In T cells from early allo-BMT patients (<4 months), various sizes of highly tyrosine phosphorylated proteins were observed as high background even without the stimulation through VLA/beta1 integrin. The high tyrosine phosphorylation pattern gradually disappeared and it was finally returned to normal tyrosine phosphorylation patterns by 2 years after BMT. Interestingly, poor expression of focal adhesion kinase (pp125FAK), a VLA/beta1-mediated signaling molecule, was observed within 1 year after BMT. These results suggest that these molecular defects appear to be implicated in the impaired VLA/beta1-mediated signaling in T cells from patients after allo-BMT, and it could explain, in part, the persistent immunoincompetent state after allo-BMT at least 1 year. PMID- 9354696 TI - Wilms tumor gene (wt1) mRNA is equally expressed in blast cells from acute myeloid leukemia and normal CD34+ progenitors. PMID- 9354697 TI - International prognostic scoring system and other prognostic systems for myelodysplastic syndromes. PMID- 9354698 TI - A cheaper and more rapid polymerase chain reaction-restriction fragment length polymorphism method for the detection of the HLA-H gene mutations occurring in hereditary hemochromatosis. PMID- 9354699 TI - Neonatal screening for the hemochromatosis defect. PMID- 9354700 TI - A prospective study of radiation therapy-associated thrombocytopenia. PMID- 9354701 TI - Cloned ligand-gated channels activated by extracellular ATP (P2X receptors). PMID- 9354702 TI - Preferential inhibition of Ih in rat trigeminal ganglion neurons by an organic blocker. AB - The potency and specificity of a novel organic Ih current blocker DK-AH 268 (DK, Boehringer) was studied in cultured rat trigeminal ganglion neurons using whole cell patch-clamp recording techniques. In neurons current-clamped at the resting potential, the application of 10 microM DK caused a slight hyperpolarization of the membrane potential and a small increase in the threshold for action potential discharge without any major change in the shape of the action potential. In voltage-clamped neurons, DK caused a reduction of a hyperpolarization-activated current. Current subtraction protocols revealed that the time-dependent, hyperpolarization-activated currents blocked by 10 microM DK or external Cs+ (3 mM) had virtually identical activation properties, suggesting that DK and Cs+ caused blockade of the same current, namely Ih. The block of Ih by DK was dose dependent. At the intermediate and higher concentrations of DK (10 and 100 microM) a decrease in specificity was observed so that time-independent, inwardly rectifying and noninactivating, voltage-gated outward potassium currents were also reduced by DK but to a much lesser extent than the time-dependent, hyperpolarization-activated currents. Blockade of the time-dependent, hyperpolarization-activated currents by DK appeared to be use-dependent since it required hyperpolarization for the effect to take place. Relief of DK block was also aided by membrane hyperpolarization. Since both the time-dependent current blocked by DK and the Cs+-sensitive time-dependent current behaved as Ih, we conclude that 10 microM DK can preferentially reduce Ih without a major effect on other potassium currents. Thus, DK may be a useful agent in the investigation of the function of Ih in neurons. PMID- 9354703 TI - Functional expression and purification of histidine-tagged rat renal Na/Phosphate (NaPi-2) and Na/Sulfate (NaSi-1) cotransporters. AB - Two mammalian sodium-dependent anion-cotransporters (NaPi-2 for phosphate and NaSi-1 for sulfate) have been expressed in Sf9 insect cells using the baculovirus expression system. A histidine tag was introduced at the C-termini in order to facilitate purification by metal-affinity chromatography. Sf9 cells infected with the histidine-tagged Ni/Pi-cotransporter exhibited more than 60-fold higher sodium-dependent transport of phosphate compared to noninfected cells. Expressed Na/Pi-cotransport exhibited a Km of Pi of 0.21 mm and an apparent Km of sodium of 92 mm. Infected cells expressed a 65 kDa polypeptide as detected by Western blotting and immunoprecipitation. Sf9 cells infected with the histidine-tagged NaSi-1 or untagged NaSi-1 protein expressed sodium-dependent sulfate cotransport up to 60-fold higher compared to noninfected cells. Transport of sulfate was highly dependent on sodium exhibiting a Km of SO2-4 of about 0.3-0.4 mm and a Km of sodium of 55 mm. By Western blotting and immunoprecipitation expressed NaSi-1 proteins were detected at 55-60 kDa. These studies demonstrate that histidine tagged proximal tubular Na-dependent cotransporters for phosphate and sulfate can be expressed functionally in Sf9 cells and that the kinetic characteristics were not altered by the introduction of a histidine tag at the C-termini. Furthermore, it is demonstrated that after solubilization under denaturing conditions histidine-tagged cotransporter proteins can be purified by metal-chelate affinity chromatography. PMID- 9354704 TI - Na absorption across endometrial epithelial cells is stimulated by cAMP-dependent activation of an inwardly rectifying K channel. AB - Previous studies in our laboratory have shown that Na absorption across the porcine endometrium is stimulated by PGF2alpha and cAMP-dependent activation of a barium-sensitive K channel located in the basolateral membrane of surface epithelial cells. In this study, we identify and characterize this basolateral, barium-sensitive K conductance. Porcine uterine tissues were mounted in Ussing chambers and bathed with KMeSO4 Ringer solution. Amphotericin B (70 microM) was added to the luminal solution to permeabilize the apical membrane and determine the current-voltage relationship of the basolateral K conductance after activation by 100 microM CPT-cAMP. An inwardly rectifying current was identified which possessed a reversal potential of -53 mV when standard Ringer solution was used to bathe the serosal surface. The K:Na selectivity ratio was calculated to be 12:1. Administration of 5 mM barium to the serosal solution completely inhibited the current activated by cAMP under these conditions. In addition to these experiments, amphotericin-perforated whole cell patch clamp recordings were obtained from primary cultures of porcine surface endometrial cells. The isolated cells displayed an inwardly rectifying current under basal conditions. This current was significantly stimulated by CPT-cAMP and blocked by barium. These results together with our previous studies demonstrate that cAMP increases Na absorption in porcine endometrial epithelial cells by activating an inwardly rectifying K channel present in the basolateral membrane. Similar patch clamp experiments were conducted using cells from a human endometrial epithelial cell line, RL95-2. An inwardly rectifying current was also identified in these cells which possessed a reversal potential of -56 mV when the cells were bathed in standard Ringer solution. This current was blocked by barium as well as cesium. However, the current from the human cells did not appear to be activated by cAMP, indicating that distinct subtypes of inwardly rectifying K channels are present in endometrial epithelial cells from different species. PMID- 9354705 TI - Subunit regulation of the human brain alpha 1E calcium channel. AB - The alpha1 subunit coding for the human brain type E calcium channel (Schneider et al., 1994) was expressed in Xenopus oocytes in the absence, and in combination with auxiliary alpha2delta and beta subunits. alpha1E channels directed with the expression of Ba2+ whole-cell currents that completely inactivated after a 2-sec membrane pulse. Coexpression of alpha1E with alpha2bdelta shifted the peak current by +10 mV but had no significant effect on whole-cell current inactivation. Coexpression of alpha1E with beta2a shifted the peak current relationship by -10 mV, and strongly reduced Ba2+ current inactivation. This slower rate of inactivation explains that a sizable fraction (40 +/- 10%, n = 8) of the Ba2+ current failed to inactivate completely after a 5-sec prepulse. Coinjection with both the cardiac/brain beta2a and the neuronal alpha2bdelta subunits increased by approximately 10-fold whole-cell Ba2+ currents although coinjection with either beta2a or alpha2bdelta alone failed to significantly increase alpha1E peak currents. Coexpression with beta2a and alpha2bdelta yielded Ba2+ currents with inactivation kinetics similar to the beta2a induced currents, indicating that the neuronal alpha2bdelta subunit has little effect on alpha1E inactivation kinetics. The subunit specificity of the changes in current properties were analyzed for all four beta subunit genes. The slower inactivation was unique to alpha1E/beta2a currents. Coexpression with beta1a, beta1b, beta3, and beta4, yielded faster-inactivating Ba2+ currents than currents recorded from the alpha1E subunit alone. Furthermore, alpha1E/alpha2bdelta/beta1a; alpha1E/alpha2bdelta/beta1b; alpha1E/alpha2bdelta/beta3; alpha1E/alpha2bdelta/beta4 channels elicited whole-cell currents with steady state inactivation curves shifted in the hyperpolarized direction. The beta subunit-induced changes in the properties of alpha1E channel were comparable to modulation effects reported for alpha1C and alpha1A channels with beta3 approximately beta1b > beta1a approximately beta4 >> beta2a inducing fastest to slowest rate of whole-cell inactivation. PMID- 9354706 TI - Mechanotransduction in colonic smooth muscle cells. AB - We evaluated mechanisms which mediate alterations in intracellular biochemical events in response to transient mechanical stimulation of colonic smooth muscle cells. Cultured myocytes from the circular muscle layer of the rabbit distal colon responded to brief focal mechanical deformation of the plasma membrane with a transient increase in intracellular calcium concentration ([Ca2+]i) with peak of 422.7 +/- 43.8 nm above an average resting [Ca2+]i of 104.8 +/- 10.9 nM (n = 57) followed by both rapid and prolonged recovery phases. The peak [Ca2+]i increase was reduced by 50% in the absence of extracellular Ca2+, while the prolonged [Ca2+]i recovery was either abolished or reduced to less than or = 15% of control values. In contrast, no significant effect of gadolinium chloride (100 microM) or lanthanum chloride (25 microM) on either peak transient or prolonged [Ca2+]i recovery was observed. Pretreatment of cells with thapsigargin (1 microM) resulted in a 25% reduction of the mechanically induced peak [Ca2+]i response, while the phospholipase C inhibitor U-73122 had no effect on the [Ca2+]i transient peak. [Ca2+]i transients were abolished when cells previously treated with thapsigargin were mechanically stimulated in Ca2+-free solution, or when Ca2+ stores were depleted by thapsigargin in Ca2+-free solution. Pretreatment with the microfilament disrupting drug cytochalasin D (10 microM) or microinjection of myocytes with an intracellular saline resulted in complete inhibition of the transient. The effect of cytochalasin D was reversible and did not prevent the [Ca2+]i increases in response to thapsigargin. These results suggest a communication, which may be mediated by direct mechanical link via actin filaments, between the plasma membrane and an internal Ca2+ store. PMID- 9354707 TI - A novel large conductance, nonselective cation channel in pheochromocytoma (PC12) cells. AB - A new type of nonselective cation channel was identified and characterized in pheochromocytoma (PC12) cells using inside-out and cell-attached patch-clamp recordings. The channel shows a large unitary conductance (274 pS in symmetric 145 mm K+) and selectivity for Na+ approximately K+ > Li+, and is practically impermeable to Cl-. The channel activity-voltage relationship is bell-shaped, showing maximal activation at approximately -10 mV. The overall activity of this channel is unmodified by [Na+]ic, or [Ca++]ic. However, increases in [Ca++]ic lead to a decrease in the unitary current amplitude. In addition, overall activity is mildly increased when suction is applied to the back of the patch pipette. Together, these characteristics distinguish the present channel from all other large conductance nonselective cation channels reported so far in a variety of preparations. The frequency of appearance of this channel type is similar in undifferentiated and NGF-treated PC12 cells ( approximately 8-27% of patches). The combination of large conductance, permeability to Na+, and existence of conducting states at negative potentials, may provide a significant pathway for inward current and depolarization in PC12 cells. PMID- 9354708 TI - Evaluation of directional atherectomy studied by intravascular ultrasound in femoropopliteal artery stenosis. AB - PURPOSE: To evaluate the role of intravascular ultrasound (IVUS) before and after directional atherectomy (DA) in the treatment of femoropopliteal artery stenosis. METHODS: In 12 patients with 16 stenoses IVUS was performed before and immediately after an angiographically successful DA. This was defined as a diameter reduction (DR) < or = 50%, which was calculated using the minimal lumen diameter compared with the diameter of a nearby "normal" segment. In the presence of residual plaque on IVUS an additional DA was performed. Endpoints studied were DR < or = 30% on IVUS compared with the IVUS findings of the angiographically normal reference segment, or when no additional atherosclerotic material could be removed by further DA passages. RESULTS: Additional DA (mean 1.6 per lesion) had to be performed in all patients. Initial DA increased the cross-sectional free lumen area (FLA) from 3.8 +/- 2.0 mm2 to 8.1 +/- 2.7 mm2 (p = 0.0004). Additional DA increased FLA to 9.3 +/- 2.3 mm2 (p = 0.002) after the second passage and to 9.8 +/- 2.4 mm2 (p = 0.09) after the final DA run. The plaque area (PLA) before DA decreased from 18.1 +/- 4.2 mm2 to 15.4 +/- 4.8 mm2 (p = 0.002) after the first passage, and to 13.5 +/- 5.0 mm2 (p = 0.004) and 12. 8 +/- 4.4 mm2 (p = 0.07) after the second and final DA runs, respectively. PLA of the reference segment (9.5 +/- 5.7 mm2) was significantly smaller (p = 0.006) than the final PLA of the treated lesion, indicating a large amount of retained plaque. As a result of DA there was an increase in the area bordered by the medial layer, i. e., the total vessel area (from 21.9 +/- 4.7 mm2 to 23.0 +/- 4.7 mm2), significantly in eccentric and soft lesions. On IVUS, dissection and plaque rupture after the final passage was seen in 12 of 16 stenoses; two dissections were seen on the completion angiogram. After the final passage in all stenoses except three, the DR with IVUS was < or = 30%. CONCLUSION: Lumen enlargement following DA is predominantly due to plaque excision. Vessel expansion combined with plaque excision varies in different stenoses and is an important factor in eccentric and soft lesions. Despite additional DA considerable plaque remains. PMID- 9354709 TI - Balloon angioplasty combined with primary stenting versus balloon angioplasty alone in femoropopliteal obstructions: A comparative randomized study. AB - PURPOSE: To evaluate whether balloon angioplasty combined with stenting (ST) of symptomatic femoropopliteal disease would provide better results compared with balloon angioplasty alone (BA). METHODS: Fifty-one patients were randomized between ST (24 patients) and BA (27 patients). Follow-up comprised clinical and hemodynamic assessment and color-flow duplex ultrasound examinations. RESULTS: Residual stenosis (> or = 30% diameter reduction) occurred in three BA patients, but not in the ST patients. By life-table analysis the cumulative rate of clinical and hemodynamic success after 1 year with ST was 74% (SE 9%) and for those with BA 85% (SE 7%) (p = 0.25). The primary patency at 1 year assessed by color-flow duplex ultrasound was 62% (SE 9%) for ST-treated patients and 74% (SE 8%) for BA patients (p = 0.22). Occlusion occurred in five ST patients (21%) compared with two BA patients (7%). CONCLUSION: ST does not improve clinical and hemodynamic outcome compared with BA. Moreover, the occlusion rate in ST-treated patients is higher. PMID- 9354710 TI - Therapeutic consequences of variation in intraarterial pressure measurements after iliac angioplasty. AB - PURPOSE: To assess the accuracy of intraarterial measurement of transstenotic pressure gradients for the detection of hemodynamically suboptimal iliac angioplasty. METHODS: In 14 patients, referred for diagnostic angiography, mean pressure gradients in the aorta and iliac artery were obtained twice, using a double-sensor pressure catheter. Additional iliac measurements were performed during pharmacologically induced flow augmentation. Repeatability was assessed by calculation of the mean difference plus standard deviation (MD +/- SD) and repeatability coefficient (2 x SD). These results were extrapolated to 137 iliac angioplasty procedures with secondary stenting where there was a residual pressure gradient > 10 mmHg. RESULTS: MD +/- SD for repeated measurements at rest and during flow augmentation were 0 +/- 2 mmHg and 1 +/- 3 mmHg, respectively. Repeatability coefficients were 3 and 6 mmHg. Mean pressure gradients after hemodynamically insufficient angioplasty were 8 +/- 7 mmHg at rest and 17 +/- 5 mmHg following vasodilatation. Inaccurate pressure recordings may have led to inappropriate stent placement in less than 2.5%, and inappropriate denial of stent placement in less than 5% of the lesions. CONCLUSION: Variability of intraarterial pressure measurements has little consequence in the detection of hemodynamically significant stenosis after angioplasty. PMID- 9354711 TI - Malignant gastric and duodenal stenosis: palliation by peroral implantation of a self-expanding metallic stent. AB - PURPOSE: To assess the use of self-expanding metallic stents in patients with inoperable malignant antrum-pylorus-duodenal obstruction. METHODS: Six patients underwent implantation of a Wallstent self-expanding metallic endoprosthesis (20 mm in five patients and 16 mm in one). In five patients a catheter (Berenstein) was introduced perorally into the stomach. A guidewire (Terumo) was introduced through the catheter and advanced through the antrum-pylorus-duodenal stenosis. The guidewire was removed and a 260-cm-long, 0.035" superstiff guide (Amplatz) was introduced. After the catheter was removed the stent assembly was introduced. In the last patient the stent was implanted through a percutaneous gastrostomy. RESULTS: Treatment of inoperable gastric outlet obstruction caused by tumor compression is difficult and unsatisfactory. Peroral implantation of self expanding metallic stents resulted in successful palliative therapy of antrum pylorus-duodenal stenosis in six patients in whom surgery was not possible because of advanced disease and poor general condition. On average, patients were able to eat during 41 days. One patient is tolerating oral intake at 3 months. CONCLUSION: Implantation of stents resulted in palliative relief of malignant antrum-pylorus-duodenal obstructions. PMID- 9354714 TI - In vitro evaluation of a rheolytic thrombectomy system for clot removal from five different temporary vena cava filters. AB - PURPOSE: To evaluate the feasibility of thrombus removal from temporary vena cava filters using a rheolytic thrombectomy device and to assess the embolization rate of this procedure. METHODS: Five temporary vena cava filters together with porcine thrombi were placed in a vena cava flow model (semitranslucent silicone tube of 23 mm diameter, pulsatile flow at a mean flow rate of 4 L/min). A rheolytic thrombectomy system (Hydrolyser) was used with a 9 Fr guiding catheter to remove the clots. The effluent was passed through filters of different size and the amount of embolized particles as well as the remaining thrombus were measured. RESULTS: Thrombus removal rates ranged from 85% to 100%. Embolization rates between 47% and 60% were calculated for the different filters. CONCLUSION: The Hydrolyser is able to remove sufficiently high amounts of thrombus from temporary vena cava filters. However, the amount of embolized particles makes it impossible to utilize this method without special precautions against embolization. PMID- 9354713 TI - Clinical experience with covered wallstents for biliary malignancies: 23-month follow-Up. AB - PURPOSE: To evaluate the effectiveness of partially covered metallic Wallstents to prevent tumoral ingrowth in patients with neoplastic obstruction of the biliary tract. METHODS: Twenty-one patients with malignant obstructive jaundice have been treated with Wallstents partially covered with a polyurethane polymer. In total, 36 covered stents (8 and 10 mm in diameter, 70 and 90 mm long) were deployed. All the stents were free from covering at both ends. RESULTS: Jaundice was successfully treated in 100% of cases. There were no problems related to the releasing system during stent positioning, no major complications, and no incompatibility reactions to the materials composing the endoprostheses. At 23 month follow-up, 6 patients are still alive and 15 are dead; of these 15 patients, 11 died in the first 6 months and the last 4 died between 6 and 23 months. Seven patients had an obstructed stent; in four of these, cholangioscopy showed the presence of tumoral ingrowth and in one it showed necrotic tissue with biliary pigments and inflammatory cells. No biopsy specimen was obtained in the remaining two patients with stent obstruction. The follow-up, ranging from 7 to 23 months, showed a primary patency of 46.8% and 24.6% and an assisted patency of 66.3% and 59% at 6 months and 23 months, respectively. CONCLUSIONS: Covered metallic stents are effective and may produce improved survival in patients with malignant biliary obstruction (27. 8% at 23 months). Stent patency, however, is similar to that of uncovered stents. Modifications in the design of the covering membrane may reduce stent obstruction resulting from disruption of the plastic covering. PMID- 9354712 TI - Transcatheter embolization of pseudoaneurysms complicating pancreatitis. AB - PURPOSE: To evaluate the therapeutic role of angiography in patients with pseudoaneurysms complicating pancreatitis. METHODS: Thirteen symptomatic pseudoaneurysms were treated in nine patients with pancreatitis. Eight patients had chronic pancreatitis and pseudocyst and one had acute pancreatitis. Clinical presentation included gastrointestinal bleeding in seven patients and epigastric pain without bleeding in two. All patients underwent transcatheter embolization. RESULTS: Transcatheter embolization resulted in symptomatic resolution in all patients. Rebleeding occurred in two patients, 18 and 28 days after embolization respectively, and was successfully treated by repeated embolization. One patient with severe pancreatitis died from sepsis 28 days after embolization. Follow-up was then available for eight patients with no relapse of bleeding after a mean follow-up of 32 months (range 9-48 months). CONCLUSION: Transcatheter embolization is safe and effective in the management of pseudoaneurysms complicating pancreatitis. PMID- 9354715 TI - The precise determination of vascular lumen and stent diameters: correlation among calibrated angiography, intravascular ultrasound, and pressure-fixed specimens. AB - PURPOSE: Luminal diameters measured in vivo by calibrated-catheter angiography and by intravascular ultrasound were correlated with those obtained from pressure fixed histologic cross-sections to determine the accuracy of both methods. METHODS: Angiographic and endosonographic diameter measurements were performed in the center of stents placed in the iliac arteries of 10 miniature pigs and were compared with luminal and stent diameters in postmortem, pressure-fixed, histologic cross-sections from identical locations. RESULTS: Compared with histologic diameters, magnification-corrected angiographic measurements still magnified vascular luminal diameters by 0.7 +/- 0.71 mm (r = 0.41, Pearson; p < 0.003, Wilcoxon, matched pairs), whereas intravascular ultrasound measurements proved to be almost identical to the histologic lumina (r = 0.95, Pearson; p > 0. 5, Wilcoxon, matched pairs). Similarly, stent diameters correlated well between endosonographic and histologic measurements (r = 0.91; p = 0.002), and less well between angiographic and histologic diameters (r = 0.62; p = 0.002). CONCLUSION: Since calibrated angiography still overestimates vascular lumina, endosonography is the preferred technique for accurate in vivo measurements. PMID- 9354716 TI - Transjugular intrahepatic portosystemic shunt: histologic and immunohistochemical study of autopsy cases. AB - PURPOSE: To assess the histologic findings associated with stenosed and occluded transjugular intrahepatic portosystemic shunt (TIPS) tracts. METHODS: Four TIPS tracts within three autopsy livers were histologically studied for vascular components by routine staining and immunohistochemical staining. TIPS had been performed for bleeding from esophageal varices in patients with cirrhosis of the liver. RESULTS: Two TIPS, examined on days 4 and 53, showed occlusion by fibrin thrombus. In the former, no endothelial cells were detected, but coagulative necrosis of hepatocytes was found in the surrounding liver. In the latter, bile pigments were seen on the luminal surface. In the two other TIPS without tract occlusion, examined on days 49 and 293, a layer of endothelial cells, proliferation of smooth muscle cells, and deposition of an extracellular matrix such as collagen were confirmed. In the tract examined on day 293, there was protrusion of hepatocytes into the lumen through the stent wires. CONCLUSION: Short- and midterm TIPS occlusions were caused by thrombus forming after necrosis of hepatocytes and bile leakage, respectively. Long-term TIPS stenosis was associated with a combination of pseudointimal hyperplasia and ingrowth of hepatocytes. PMID- 9354717 TI - A dedicated Z-stent for acquired saber-sheath tracheobronchomalacia. AB - The tracheobronchial lumen has a continuous horseshoe arch morphology. We formed Z-stents accordingly to support the weakened cartilagenous portions. With this type of stent we treated a patient with acquired saber-sheath type tracheobronchomalacia (TBM), Rayl's type II, Johnson's grade III, whose condition was aggravated even under positive end expiratory pressure (PEEP) therapy. The patient improved gradually. No immediate complication was observed. Bronchofiberscopic examination revealed that the tracheobronchial arcade was closely strut-braced and showed no expiratory collapse. Six months later, when the patient was intubated due to asthmatic attacks, tissue ingrowth through the stent was found and removed. There was no recurrence of TBM. The patient died 2 years later of pneumoconiosis. PMID- 9354718 TI - Transluminal treatment of a celiac artery pseudoaneurysm with a stent graft occlusion device. AB - This report describes the transluminal placement of a stent graft occlusion device to treat a celiac bypass graft pseudoaneurysm which was causing biliary and duodenal obstruction. PMID- 9354719 TI - Perforation of the esophagus secondary to insertion of covered wallstent endoprostheses. AB - Perforation of the esophagus is a very rare complication of metallic esophageal stent insertion. Two cases are presented in which esophageal perforations were caused by the sharp ends of metallic stents impinging on the esophageal wall. In retrospect, both perforations might have been prevented by additional stent insertion. PMID- 9354720 TI - Dissection of the abdominal aorta in blunt trauma: management by percutaneous stent placement. AB - We implanted stents in three patients who had traumatic abdominal aortic dissections, complicated by right limb ischemia in one case. The circulating false channel extended to the left iliac artery in one case and to both iliac arteries in the last case. Diagnosis and radiological follow-up included ultrasound, computed tomography, and arteriography. Two patients were treated with Wallstents, one with a Palmaz stent. The occlusion of the false channel was obtained in all patients without any significant residual stenosis. No early or late complication was noted in any of the patients. The longest follow-up was 2 years. We conclude that stent placement is an efficient method for the treatment of noniatrogenic inframesenteric aortic dissections. PMID- 9354721 TI - Retrieval of migrated colonic stents from the rectum. AB - Palliative stenting of malignant colonic obstruction may be complicated by stent migration. Stents that migrate into the rectum cannot be passed with bowel movements and frequently cause obstruction. We present two simple means to retrieve stents from the rectum using fluoroscopic guidance. These techniques were used successfully without complication in four stent migrations. PMID- 9354723 TI - The effects of Down syndrome and other chromosomal abnormalities on survival and management in oesophageal atresia. AB - Recognisable chromosomal abnormalities occur in over 5% of patients with oesophageal atresia (OA). In a review of 670 patients with OA chromosomal abnormalities were identified in 35 (5.2%), of whom 16 had trisomy 18 and 12 had trisomy 21. In patients with trisomy 18, the diagnosis should be suspected on clinical grounds and confirmed on analysis of chromosomes; no active treatment of the OA is justified because of the extremely poor prognosis. In Down syndrome (DS) 50% will have pure OA with no tracheo-oesophageal fistula. In addition, many of these infants will have associated anomalies typical of those normally seen in DS, eg., Hirschsprung's disease, duodenal atresia, and congenital heart disease. Despite treatment, OA with DS has a high mortality. PMID- 9354724 TI - Surface electrogastrography in children with esophageal atresia. AB - Surface electrogastrography was performed in 18 patients with esophageal atresia (EA) and 10 normal controls to investigate the possible role of a congenital enteric nerve defect as a cause of gastroesophageal reflux (GER), which is common after repair of EA. The means of the dominant frequencies and ranges of the frequency distribution were compared. The dominant frequencies (0.047+/-0.007 Hz) in the EA group did not differ significantly from those of the controls (0.050+/ 0.007 Hz, P >0.1), although 2 patients had bradygastria and 2 had tachygastria in the EA group. The range of the frequency distribution was significantly wider in the EA group compared with normal children (P = 0.002). The wide frequency distribution in children with EA suggests disturbed electrical activity of the stomach, which could be associated with poor electromechanical coupling and, hence abnormal gastric contraction. PMID- 9354725 TI - Cardiovascular malformations in rat fetuses with oesophageal atresia and tracheo oesophageal fistula induced by adriamycin. AB - Associated congenital anomalies have emerged as the most significant prognostic factor in babies born with oesophageal atresia and/or tracheo-oesophageal fistula (OA-TOF). The most frequently encountered groups of anomalies are cardiovascular (CV) and gastrointestinal, the former being more significant from a prognostic point of view. Some, such as a right-sided aortic arch (RAA), vascular ring, or major heart defects, may alter the timing and surgical approach for the repair of OA-TOF or adversely affect the prognosis. The rat fetal OA model induced by adriamycin (Adr) has been described previously. In the present experiments, information was sought regarding the incidence and type of CV abnormalities in fetal rats obtained from Adr-treated dams. OA-TOF was induced in 24 of 36 fetal rats from Adr-treated dams. DV abnormalities were found in 18 (75%) OA-TOF fetuses and 10 (83%) Adr-treated fetuses without OA-TOF. The difference was not significant (P >0.05). The most frequently found anomalies were ventricular and atrial septal defects. A RAA was present in 8/36 fetuses and a double aortic arch in 2/36. A patent ductus arteriosus was present in all treated fetuses compared with two-thirds of controls. The findings in the present study emphasise the importance of CV anomalies in association with OA, and reinforce the value of the Adr-induced rat fetal OA model by adding to our knowledge of the basic embryogenesis of both OA and CV anomalies. PMID- 9354727 TI - Esophageal manometric studies in children with achalasia before and after operative treatment. AB - This study manometrically assessed and compared esophageal function in 16 children with achalasia before and after surgical treatment (anterior esophagomyotomy with antireflux partial fundoplication). Manometric examinations were done in 10 children preoperatively and in 12, 3 months to 8 years postoperatively. Both pre- and postoperative examinations were done in 6 patients. The following parameters were measured: lower esophageal sphincter (LES) pressure and length, spontaneous motility of the esophageal body, and motility provoked by swallowing of fluids. Preoperative examinations confirmed disturbances typical for achalasia: increased LES pressure (mean 39.4 mmHg), lack of relaxation upon swallowing, and various types of anomalous esophageal motility (lack of propulsive waves, segmental waves, breaks in propagation of contractions, tonic contractions, etc.). Postoperative examinations showed normalization of LES pressure; however, relaxation did not appear in any patient. Esophageal motility improved after surgery in most patients and was already noticeable 3-6 months postoperatively, but motility never returned to normal. Clinically, all but 1 patient with reflux esophagitis were doing well despite persistent motility disturbances. Our study confirms that achalasia is a complex motor disorder of the entire esophagus. The improvement of esophageal contractility after esophagomyotomy suggests both primary and significant secondary damage to motility of the esophageal body in most patients. It appears that secondary disturbances are reversible to some extent in children after surgical treatment. PMID- 9354726 TI - Additional congenital anomalies in babies with gut atresia or stenosis: when to investigate, and which investigation. AB - A wide variety of additional congenital anomalies occur in babies born with a gut atresia or stenosis. The specific pattern of anomalies depends on the location of the atresia. The serious nature of many of them makes perioperative diagnosis imperative. Eighty-six babies born with pure oesophageal atresia (OA), duodenal atresia (DA) or stenosis, or jejuno-ileal atresia (JIA) have been studied. These, combined with over 2,000 cases in the literature, have been used to develop a protocol to optimally investigate babies with gut atresia for associated anomalies. The authors recommend routinely obtaining anterio-posterior and lateral chest and abdominal radiographs for babies with pure OA, DA and intestinal atresia, making sure the entire spine can be visualised. Cardiac and renal ultrasonography (US) should be routine in all babies with pure OA or DA. A micturating cystourethrogram should be done in those babies with abnormal urinary tract US or an associated anorectal anomaly. A sweat test should be obtained in babies with JIA, and a rectal biopsy should be taken in babies with the combination of Down's syndrome and DA to exclude Hirschsprung's disease. PMID- 9354728 TI - Altered messenger RNA expression of the neuronal nitric oxide synthase gene in infantile hypertrophic pyloric stenosis. AB - Nitric oxide (NO) has been described as a mediator of smooth muscle relaxation in the mammalian gastrointestinal tract. The enzyme neuronal nitric oxide synthase (NOS) catalyzes the formation of NO. We examined the expression of the neuronal NOS gene at the messenger RNA (mRNA) level in pyloric smooth-muscle biopsy specimens from six patients with infantile hypertrophic pyloric stenosis (IHPS) using a reverse transcription-polymerase chain reaction (RT-PCR) technique. For controls, smooth-muscle layer specimens of pylorus (n=3), ileum (n=2), and colon (n=2) were used. With 31 cycles of PCR reaction, control specimens revealed detectable signals for neuronal NOS mRNA. In contrast, signals of IHPS specimens were undetectable in five cases and very weak in one. By increasing the PCR to 37 cycles, detectable signals for neuronal NOS mRNA were observed in all IHPS specimens, but they were significantly weaker than those of controls. Since a low level of neuronal NOS mRNA may lead to impaired production of NO, our observations indicate that the excessively contracted, hypertrophied pyloric muscle in IHPS is a result of reduced expression of the neuronal NOS gene at the mRNA level. PMID- 9354729 TI - Are babies with gastroschisis small for gestational age? AB - A large proportion of babies with gastroschisis (GS) have low birth weights. It is not clear, however, whether only certain subgroups or the whole population of babies with GS have low birth weights. The aim of this study was to ascertain if the birth weights of babies with GS are significantly lower than those of the general population and to determine if the birth weights of babies with GS from two different populations were significantly different. From 1969 to 1995, 44 babies with GS were treated at Auckland Children's Hospital, New Zealand. From 1980 to 1993, 69 babies were treated at Birmingham Children's Hospital, England. For each group, the mean birth weight relative to the mean birth weight for gestation (WtStdev) was significantly different from zero (Auckland = -0.806, Birmingham = -0.762, P < 0.001, one-sample analysis). The mean WtStdev scores from each centre were not significantly different from each other. Our data demonstrate that the birth weights of babies with GS are significantly lower than those of the general population and are similar in different populations. These findings support the notion that a normally functioning intestinal tract is essential for normal fetal growth. PMID- 9354730 TI - End-results of experimental gastroschisis created by abdominal wall versus umbilical cord defect. AB - An experimental study was conducted to determine the end-results of two different defects on the anterior abdominal wall: an abdominal wall defect (AWD) versus an umbilical cord defect (UCD) using chick embryos. The AWD was created by leaving an intact skin bridge between the defect and the umbilical cord in group 1; the UCD was created on the umbilical cord near the junction of the skin in group 2. At the end of incubation, the intestines appeared hemorrhagic in the AWD group, but not in the UCD group. During microscopic examination, hemorrhagic areas were observed in the bowel wall and mucosal villi in the AWD group but not in the UCD group. The end-result of the defect causing the physiological umbilical hernia resulted in bowel damage resembling the classic picture of gastroschisis (GS). We conclude that the site of the defect in GS is not the abdominal wall itself, but the physiological umbilical hernia. PMID- 9354731 TI - Laparoscopic versus open cholecystectomy in children. AB - Twenty-one consecutive laparoscopic cholecystectomies (LC) were compared with 29 consecutive open cholecystectomies (OC). Sickle-cell disease (SCD) was the most common reason for cholecystectomy in both groups. The average length of operative time for LC was significantly longer than that of OC (P=0.0149). In 1 patient there was conversion from LC to OC due to severe adhesions. Common bile duct (CBD) stones were diagnosed in 8 (27.6%) of the OC group; in 4 of them the diagnosis was made preoperatively by ultrasound, in 4 by intraoperative cholangiogram. All 8 patients required CBD exploration, and 2 had additional transduodenal sphincteroplasties. In the LC group 5 patients (23.8%) had CBD stones. All had (ERCP) endoscopic retrograde cholangiopancreatography sphincterotomy, and stone extraction followed by LC. ERCP is a necessary adjunct to treatment if LC is to be contemplated. Six patients in the OC group developed complications, while only 4 patients in the LC group developed minor complications. The length of hospitalization after LC was significantly shorter than after OC (P=0.0150). LC is the procedure of choice in the management of cholelithiasis in children, especially those with SCD. PMID- 9354732 TI - Does treatment with human chorionic gonadotropin induce reversible changes in undescended testes in boys? AB - Between May 1993 and November 1995, 71 cryptorchid boys were treated with human chorionic gonadotropin (hCG); 42 were operated upon following unsuccessful hCG treatment. A routine orchiopexy was performed in each case. In 10 cases a testicular biopsy was made during orchipexy within 3 days following hCG treatment; in another 10 biopsies were taken 6 to 9 months after treatment. Testicular biopsies were taken at the time of orchiopexy in 5 cryptorchid boys who were not treated with hCG as a control group. A mild, inflammation-like reaction was found in the cryptorchid testes in the period immediately following the last hCG injections, but those studied 6 to 9 months after the last injection there were no apparent such reactions. In contrast to the inflammation-like reaction, the volume density of blood vessels, interstitial bleeding, and diameter of the seminiferous tubules had not regressed. The numbers of spermatogonia per tubular transverse section and the percentage of tubular transverse sections containing spermatogonia (the fertility index) were increased. PMID- 9354733 TI - Pediatric pheochromocytoma. A 36-year review. AB - Fourteen children (10 boys and 4 girls, aged 8 to 17 years) had 20 pheochromocytomas treated over a 36-year period from 1959 to 1995 inclusive. Nine patients had 11 tumors before 1980; 5 children had 9 tumors up to 1987. There were no new children with pheochromocytomas at our hospital from 1988 to 1995. Hypertension, sweating, headache, and visual blurring were the most common symptoms and signs (average 5 months). The most reliable biochemical investigations were the urinary catecholamines and norepinephrine. Before 1980, intravenous pyelography and angiography were most successful in localizing the tumor, but since then ultrasonography and computerized tomography have been the radiological investigations of choice. Early involvement of the anesthesiologist in the preoperative control of the hypertension is essential; blood pressure (BP) control was achieved with phenoxybenzamine. The main anesthetic drugs used were: sodium thiopental, fentanyl, methoxyflurane, isoflurane, nitrous oxide, and metocurine. Sixteen tumors were adrenal and 4 were extra-adrenal (1 intrathoracic and 1 extradural). All except 2 tumors were completely resected; they ranged in size from 1.3 to 14 cm. Ligation of the tumor's venous drainage was usually associated with a sudden, temporary fall in systemic BP. There were 2 children with malignant tumors. Four patients had five recurrences (second pheochromocytoma) within 6 years, and all were heralded by a return of their original symptoms and signs. One girl was left with no adrenal tissue. The only complication was in a boy with a large, partly-resected malignant right adrenal tumor who had a subphrenic abscess drained and was left with a temporary bile fistula, cirrhosis, and chronic pain. All children were normotensive when discharged from hospital and remain alive and well with a follow-up of 7 to 36 years. There were no deaths. Long-term follow-up is essential. Key word Pheochromocytoma PMID- 9354734 TI - Actual half-life of alpha-fetoprotein as a prognostic tool in pediatric malignant tumors. AB - In a retrospective study, the prognostic value of monitoring the decay of alpha fetoprotein (AFP) was assessed. Serum AFP was determined serially in 18 children with malignant germ-cell or hepatic tumors: 7 endodermal sinus tumor, 3 embryonal carcinoma, 5 malignant teratoma, 2 hepatoblastomas, and 1 hepatocellular carcinoma. The actual half-life (AHL) of AFP was computed after surgical resection of the tumor. In group 1, which had complete resection and no recurrence during follow-up (n = 13), the AHL of AFP was 4.0 +/- 0.9 days. In group 2, which had incomplete resection or recurrence during follow-up (n = 5), the AHL of AFP was 24.8 +/- 20 days, significantly longer than that of group 1 (P = 0.0026). The increased AHL of AFP indicated residual active tumor after surgical resection. The AHL of AFP may be more sensitive than serial monitoring of AFP in detecting preclinical recurrence after surgical resection of AFP secreting tumors. Treatment strategies can be based on AFP clearance, and prospective clinical trials are warranted. PMID- 9354735 TI - Ureteral fibroepithelial polyps in children. AB - Fibroepithelial polyps of the ureter presenting as pelviureteric junction (PUJ) obstruction in two boys are reported. These neoplasms are uncommon, especially in children. Surgical excision of the PUJ with the polyp and dismembered pyeloplasty was performed in each case. Postoperative recoveries were uneventful. PMID- 9354736 TI - Combined rectal atresia and rectal stenosis. AB - A unique case of combined rectal atresia and rectal stenosis with a normal anal canal is reported. To our knowledge, no similar case has been reported to date. PMID- 9354737 TI - Successful separation of thoraco-omphalopagus and ischiopagus tetrapus twins in Korea. AB - A pair of male thoraco-omphalopagus twins with common liver, diaphragm, pericardium, and sternum was separated at the age of 59 days after a parasitic relationship had developed between them. Before separation one baby developed acute renal failure during which he had no edema and had normal serum electrolytes, urea nitrogen, and creatinine due to "autodialysis" by the other baby. The boys have now grown normally to the age of 6 years. A pair of female ischiopagus tetrapus twins who had a common terminal ileum and colon with imperforate anus was separated at the age of 20 h. The smaller baby had congenital multiple arthrogryposis of both lower extremities, a fracture at the middle of the left femur, and a double vagina with hydrocolpos due to an imperforate hymen of the right vagina and a rectovaginal fistula on the left. Posterior sagittal rectoplasties were performed at 7 months of age in both babies. They have normal bowel movements. All four children are alive and developing normally. These are the first two case reports of successfully separated conjoined twins in Korea. PMID- 9354738 TI - Management of perineal canal anomaly. AB - A 6-year-old girl presented due to passage of stool through her vulva since birth. Examination revealed a fistulous tract between vestibule of the vagina and an otherwise normally formed anal canal. The tract was successfully excised through an anterior sagittal approach with a defunctioning sigmoid colostomy, which was closed 12 weeks later. The embryology, morbid anatomy, and treatment of this rare congenital anomaly are discussed along with a review of the literature. PMID- 9354739 TI - Recurrent mediastinal mass in a child with Hodgkin's disease following successful therapy: a diagnostic challenge. AB - The case of an 11-year-old girl with mediastinal stage III B-E Hodgkin's disease is described. She achieved complete remission with combined chemoradiotherapy according to the Swiss Pediatric Oncology Group-HD Protocol 1985. Six months after all therapy was stopped, a slowly growing retrosternal mass was detected. Computed tomography (CT) and gallium-67 single-photon emission CT (SPECT) could not elucidate the true origin of the tumor, nor did ultrasound-guided transthoracic fine-needle puncture. Open biopsy with histologic examination of the lesion has successfully identified the mass as thymic hyperplasia, a rebound immunologic reaction after chemoradiotherapy that mimicked tumor regrowth. PMID- 9354741 TI - Long-gap oesophageal atresia. PMID- 9354740 TI - Ductoplasty for an aberrant hepatic duct in a choledochal cyst. AB - The confluence of the right and left hepatic ducts at the hepatic hilum frequently shows normal anatomic variations. Choledochal cysts (CC) are also accompanied by similar variations, and devices for free drainage of bile are occasionally required in biliary reconstruction. We present a CC that had an aberrant posterior branch of the right hepatic duct draining into the distal common hepatic duct. A capacious hepaticoduodenostomy at the hilum was performed after joining the hilar and aberrant ducts. PMID- 9354742 TI - Concomitant simple repair of pectus excavatum associated with tetralogy of Fallot. PMID- 9354743 TI - Unusual case of testicular ectopia. PMID- 9354744 TI - Failure to thrive, sacropenia and functional decline in the elderly. AB - At this time, there are no commonly accepted definitions for either the "failure to thrive" syndrome or for the newly described syndrome of sarcopenia. This lack of a common definition makes it difficult to contrast study results and understand the etiology of these syndromes. This article reviews the current state of understanding of these conditions and suggests potential mechanisms that may be further investigated. PMID- 9354745 TI - Neuropsychiatric aspects of failure to thrive in late life. AB - Both depression and dementia can lead to failure to thrive (FTT). Depression can lead to FTT by two routes: a direct path related to decreased appetite as a symptom of depression; and an indirect path related to the effect of depression in increasing disability. Depression associated with FTT should usually be treated with antidepressant medication. In Alzheimer's patients with FTT, the thrust of treatment is the identification and treatment of the medical and psychiatric comorbidities and the appropriate titration of environmental supports. PMID- 9354746 TI - Functional, social, and psychological disability as causes of loss of weight and independence in older community-living people. AB - This article reviews functional, social, and psychological disabilities that relate to weight loss and independence in older community residents and suggests possible ways in which these factors may be alleviated. Although these disabilities clearly interact with one another as causes of failure to thrive, this article is organized into three major sections: functional disability, psychological and social factors, and malnutrition. In-depth geriatric assessment provides directions to reverse or halt failure to thrive. Using case materials, possible interventions are presented; these include dietary changes, a carefully planned program of physical exercise, treatment for depression, and a combination of social and environmental "prescriptions" designed to reduce social and emotional isolation. Appropriate social supports are also necessary, as is careful attention to how caregiver stress may be reduced through suitable interventions. PMID- 9354747 TI - Chronic inflammation in older people: recognition, consequences, and potential intervention. AB - In elderly individuals, some chronic inflammatory diseases appear to occur with increased frequency, and recent evidence suggests that some very common chronic, age-related disorders may be propagated and perpetuated by inflammatory processes, perhaps giving rise to the abnormal acute phase protein levels that are seen with increased frequency with aging. The consequences of chronic inflammation in the elderly undoubtedly contribute to excessive morbidity in this population. Treatment of chronic inflammation in the elderly is often difficult, requiring utmost care and close follow-up by a knowledgeable and dedicated physician. PMID- 9354748 TI - The somatopause: should growth hormone deficiency in older people Be treated? AB - Secretion of growth hormone (GH) by the pituitary gland progressively declines beginning in early adult life, a phenomenon which is termed "the somatopause." The observation that many of the changes which occur with advancing age are opposite to the physiologic effects of GH suggests that declining levels of GH and insulin-like growth factor I (IGF-I), the mediator of many of the actions of GH, may be responsible for some of these changes. This article reviews the current understanding of mechanisms underlying the somatopause, the changes of aging which may result from diminished activity of the GH-IGF-I axis, the evidence which suggests that GH "replacement therapy" may reverse these changes, and the risks of treating somatopause with GH replacement. PMID- 9354749 TI - Testosterone and frailty. AB - Although estrogen replacement therapy has become well established in the management of postmenopausal women, most physicians pay much less attention to the testosterone status of their older male patients. There is now increasing evidence that testosterone deficiency-Low Testosterone Syndrome-occurs in older men and is associated with decreased muscle strength and bone density, as well as memory problems. This article reviews the evidence for the existence of Low Testosterone Syndrome, often characterized as the male menopause or viropause, and the emerging evidence that testosterone therapy may ameliorate some of the symptoms and signs of frailty in men beyond 50 years of age. PMID- 9354751 TI - Involuntary weight loss in elderly outpatients: recognition, etiologies, and treatment. AB - Weight loss appears to occur frequently in older adults and has been associated with increased morbidity and mortality. However, the significance of weight loss per se and the optimal clinical approach to older outpatients who are losing weight is not completely understood. This article reviews the existing literature on weight loss in older people and outlines a basic approach for the clinician faced with the common, but often difficult, diagnostic and management issues presented by an elderly patient with weight loss. PMID- 9354750 TI - Immunological aspects of aging and malnutrition: consequences and intervention with nutritional immunomodulators. AB - As knowledge of molecular and cellular mechanisms of development and aging is elucidated, the need to understand the relationships among tissues, organ systems, health habits, nutrition, physical activity, and environmental factors on successful aging becomes more explicit. Progress in our understanding of how the immune system functions and responds with other factors, such as aging and nutrition, is spawning significant inroads to achieving a successful old age. PMID- 9354752 TI - Assessing and treating weight loss in nursing home patients. AB - Any physician who cares for nursing home residents has been confronted with the problem of unintentional weight loss. This can be a particularly vexing problem when the person with weight loss has multiple medical diagnoses, takes many medications, and has some degree of cognitive impairment. Questions raised in such cases include the following: what are normal weights for the old; is some degree of weight loss part of normal aging; what are the causes of weight loss in nursing home residents; and what are the most effective means of reversing the loss. PMID- 9354754 TI - Clinical evaluation of failure to thrive in older people. AB - The purpose of this article is to outline a clinical approach to patients with failure to thrive in old age. The article starts with a hypothetical clinical case presentation epitomizing the problem seen in an outpatient clinic by geriatricians. This case is then used as a basis to discuss salient features of failure to thrive which may be different from patient to patient. The focus is on using standard outpatient evaluation methods to identify common problems causing failure to thrive. PMID- 9354753 TI - Evaluating and treating nutritional problems in older patients. AB - Up to 65% of elderly hospitalized patients are protein-energy undernourished at admission or acquire nutritional deficits while hospitalized. For the elderly patient who requires nutritional support therapy, a strategy of intervention should be carefully formulated based on the results of a clinical assessment. While receiving nutritional support, the patient should be monitored carefully for complications and changing metabolic requirements. Modifications to the original regimen should be made to meet the patient's needs as necessary. PMID- 9354755 TI - Ken Michaels named 1996 Louis Schmidt Laureate. PMID- 9354756 TI - The Biological Photographic Association and professionalism revisited. PMID- 9354757 TI - Protein-nucleic acid interactions and DNA conformation in a complex of human immunodeficiency virus type 1 reverse transcriptase with a double-stranded DNA template-primer. AB - The conformation of the DNA and the interactions of the nucleic acid with the protein in a complex of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and 19-mer/18-mer double-stranded DNA template-primer (dsDNA) are described. The structure of this HIV-1 RT complex with dsDNA serves as a useful paradigm for studying aspects of nucleotide polymerases such as catalysis, fidelity, drug inhibition, and drug resistance. The bound dsDNA has a bend of approximately 41 degrees at the junction of an A-form region (first five base pairs near the polymerase active site) and a B-form region (the last nine base pairs toward the RNase H active site). The 41 degrees bend occurs smoothly over the four base pairs between the A-form portion and the B-form portion in the vicinity of helices alpha H and alpha I of the p66 thumb subdomain. The interactions between the dsDNA and protein primarily involve the sugar-phosphate backbone of the nucleic acid and structural elements of the palm, thumb, and RNase H of p66, and are not sequence specific. Amino acid residues from the polymerase active site region, including amino acid residues of the conserved Tyr Met-Asp-Asp (YMDD) motif and the "primer grip," interact with 3'-terminal nucleotides of the primer strand and are involved in positioning the primer terminal nucleotide and its 3'-OH group at the polymerase active site. Amino acid residues of the "template grip" have close contacts with the template strand and aid in positioning the template strand near the polymerase active site. Helix alpha H of the p66 thumb is partly inserted into the minor groove of the dsDNA and helix alpha I is directly adjacent to the backbone of the template strand. Amino acid residues of beta 1', alpha A', alpha B', and the loop containing His539 of the RNase H domain interact with the primer strand of the dsDNA. PMID- 9354758 TI - Mechanistic link between DNA methyltransferases and DNA repair enzymes by base flipping. AB - Rotation of a DNA nucleotide out of the double helix and into a protein binding pocket ("base flipping") was first observed in the structure of a DNA methyltransferase. There is now evidence that a variety of proteins, particularly DNA repair enzymes, use base flipping in their interactions with DNA. Though the mechanisms for base movement into extrahelical positions are still unclear, the focus of this review is how base recognition is modulated by the stringency of binding to the extrahelical base(s) or sugar moiety. PMID- 9354759 TI - Protein-DNA recognition complexes: conservation of structure and binding energy in the transition state. AB - This paper considers how enzymes that catalyze reactions at specific DNA sites have been engineered to overcome the problem of competitive inhibition by excess nonspecific binding sites on DNA. The formation of a specific protein-DNA recognition complex is discussed from both structural and thermodynamic perspectives, and contrasted with formation of nonspecific complexes. Evidence (from EcoRI and BamHI endonucleases) is presented that a wide variety of perturbations of the DNA substrate alter binding free energy but do not affect the free energy of activation for the chemical step; that is, many energetic factors contribute equally to the recognition complex and the transition-state complex. This implies that the specific recognition complex bears a close resemblance to the transition-state complex, such that very tight binding to the recognition site on the DNA substrate does not inhibit catalysis, but instead provides energy that is efficiently utilized along the path to the transition state. It is suggested that this view can be usefully extended to "noncatalytic" site-specific DNA-binding proteins like transcriptional activators and general transcription factors. PMID- 9354760 TI - Molecular and biological constraints on ligand-binding affinity and specificity. AB - Interactions between biological macromolecules have characteristic values of affinity and specificity that are set according to the biological function that is served by the interaction in the organism. Here we examine the molecular mechanisms that are used to achieve the required values of affinity and specificity in various biological systems. PMID- 9354761 TI - Sex chromosomes and sex determining mechanisms in birds. AB - The sex chromosomes in birds are designated Z and W, and the male is the homomorphic sex (ZZ) and the female heteromorphic (ZW). In most avian species the Z chromosome is a large chromosome, usually the fourth or fifth largest, and it contains almost all the known sex-linked genes. The W chromosome is generally a much smaller microchromosome, containing a high proportion of repeat sequence DNA. Recently a gene encoding a protein involved in transcriptional activation of chromatin has been detected on the W chromosome. The weight of evidence suggests that sex determination in birds is by a genic balance mechanism, in which the ratio of autosomes to Z chromosomes is the crucial factor. DNA sequences homologous to the testis determining factor in humans have been detected in both male and female birds, but it is not clear that they have a sex-related function in birds. A number of different practical methods have been developed to distinguish the sex of birds, based on sex-linked genes, the amount of DNA per cell and using DNA probes for sex-linked sequences. PMID- 9354762 TI - Bones, feathers, teeth and coat markings: a unified model. AB - A first necessary criterion to be met by any model for pattern formation is that it must be able to reproduce the patterns it purports to model. The above examples show how simple ideas from self-organisation can produce spatial patterns of varying complexity that are consistent with those observed experimentally. Second, the model must be consistent with the results of experimental manipulation. Third, the model must make experimentally testable predictions. In this way, a mathematical model can help to elucidate the underlying biochemical and biophysical mechanisms of pattern formation. I have tried to illustrate these ideas with the above examples. These examples also show the breadth of patterning phenomena that can be captured by these models. In animal coat markings, the patterns are laid down simultaneously. In limb development, the skeletal elements are laid down sequentially. The application to feather germ formation illustrates complex sequential regular pattern formation, while the application to tooth primordium formation illustrates sequential irregular pattern formation. Many other patterning phenomena have been studied (see, for example, ref. 7). Pattern formation is one of the central issues in developmental biology and intense interdisciplinary research involving experimentalists and theoreticians is beginning to help us understand how this phenomenon occurs. A detailed understanding of normal development is a necessary first step to the understanding of abnormal development and, hopefully, will help medical science combat developmental defects. PMID- 9354763 TI - Tissue factor pathway inhibitor expression in atherosclerosis. AB - Tissue factor pathway inhibitor (TFPI) is a potent inhibitor of tissue factor (TF) -initiated coagulation and may play a role in regulating coagulation in atherosclerotic plaques. The expression of TFPI protein and mRNA was examined by immunohistology and in situ hybridization in normal human and rabbit arteries, in human carotid arteries with advanced atherosclerosis, and in atherosclerotic aortas from cholesterol-fed rabbits. In normal human and rabbit arteries, TFPI protein and mRNA were detected in the adventitial layer but were undetectable in the luminal endothelium. In the medial smooth muscle layer of rabbits, weak expression of TFPI mRNA, but not protein, was detected; in that of humans, neither TFPI mRNA nor protein was detectable. In atherosclerotic arteries, TFPI protein and mRNA were detected in three of six internal carotid plaques from patients undergoing endarterectomy, and mRNA alone was detected in one further specimen. TFPI protein was found in areas of the plaque where TF was abundant and colocalized with macrophages, suggesting that these cells are responsible for TFPI synthesis. TFPI protein and mRNA were also detected in fatty-streak lesions in 18 of 19 rabbits fed a high-cholesterol diet for periods between 4 and 16 weeks. In these macrophage-rich lesions, expression of TFPI protein and mRNA was most intense at the base of the plaques. These studies suggest that TFPI is expressed in the adventitial layer of large arteries and that in atherosclerotic vessels, TFPI is expressed by macrophages in focal areas throughout the plaque. Local production of TFPI may regulate procoagulant activity and thrombotic events within atherosclerotic plaques. PMID- 9354764 TI - Identification of the circulating amyloid precursor and other gelsolin metabolites in patients with G654A mutation in the gelsolin gene (Finnish familial amyloidosis): pathogenetic and diagnostic implications. AB - Familial amyloidosis of the Finnish type (FAF) is an autosomal dominant type of systemic amyloidosis caused by a G654A (Asn-187) or G654T (Tyr-187) mutation in the gelsolin gene. Herein we show that patients with the Asn-187 gelsolin mutation have, in addition to full-sized gelsolin, a series of lower-Mr C terminal fragments of gelsolin (Mr of 70,000-45,000) in the circulation, and that a 50 to 55-kd fragment of gelsolin is excreted in the urine. In homozygous FAF (Asn-187), the 65-kd fragment, which contains the amyloid-forming region (Ala173 Met243), and the 55-kd fragment, which is devoid of that region, are the major gelsolin species in plasma; whereas normal gelsolin, as well as a 70-kd fragment identified as the C-terminal portion of gelsolin starting at Glu122, and a 45-kd fragment starting at Ser384, are minor components. In patients heterozygous for the Asn-187 mutation--the usual form of the expression of the dominant disease- normal-sized gelsolin is the major circulating form; the 65- and 55-kd fragments represent minor components. Immunodetection of the plasma 65-kd gelsolin fragment, which is disease-specific, and measurement of the urinary gelsolin fragment provide useful means for diagnosing FAF. PMID- 9354765 TI - Correlation of major histocompatibility complex with opportunistic infections in simian immunodeficiency virus-infected rhesus monkeys. AB - The role of the major histocompatibility complex (MHC) in the pathogenesis of AIDS is complex because of compounding variables within the virus, host, and environment. Important variables can be controlled by using the experimental animal model of AIDS induced by simian immunodeficiency virus in rhesus monkeys (Macaca mulatta). We studied whether the MHC type influenced which opportunistic infections arose in an individual monkey. Several associations were found. For example, cytomegalovirus was strongly associated with Mamu-B6 (p < 0.001), whereas Cryptosporidium was associated strongly with Mamu-DR3 (p < 0.001). We also found that having one opportunistic infection increased the risk of having another. PMID- 9354766 TI - Activin A: a novel player and inflammatory marker in inflammatory bowel disease? AB - Recently, we demonstrated a strong induction of activin expression after cutaneous injury. We speculated, therefore, that activin may be overexpressed during inflammatory processes in other tissues characterized by mesenchymal/epithelial structure. Herein, we show a strikingly increased expression of the activin beta A-subunit in surgical specimens from the gut of patients suffering from ulcerative colitis and Crohn's disease, whereas no activin beta A mRNA could be detected in the normal human digestive tract. The levels of activin beta A expression showed an outstanding correlation with the degree of inflammation as assessed by histologic analysis of adjacent tissue and expression analysis of the proinflammatory cytokine interleukin-1 beta. In situ hybridization studies revealed the highest levels of activin mRNA in the mucosa and submucosa of highly inflamed areas, particularly where the intestinal epithelium was damaged, but not in control tissue. In contrast, activin beta B mRNA levels in most specimens from inflamed areas were only slightly higher compared to control tissue. The strong overexpression of activin beta A in inflammatory bowel disease suggests a novel and important role of this growth and differentiation factor during inflammatory processes of the gut. PMID- 9354767 TI - Activated skin mast cells are involved in murine hair follicle regression (catagen). AB - Increasing evidence supports a role for mast cells (MC) in the control of tissue remodeling. Using the cyclic growth and regression activity of the murine hair follicle (HF) as a model, we have previously demonstrated that MC are involved in regulating the HF transformation from resting (telogen) to active hair growth (anagen). In the present study, we investigated the potential role of skin MC in spontaneous HF regression (catagen), a rapid and highly controlled process of organ involution characterized by massive epithelial cell apoptosis. By histochemistry, immunohistochemistry, and electron microscopy, we first assessed the number, location, and granulation status of perifollicular MC during the anagen-catagen-telogen transformation of back skin HF. Spontaneous catagen induction was associated with a dramatic reduction of dermal MC numbers, preceded by an increase in the percentage of degranulated MC. In vivo, the MC secretagogues substance P and adrenocorticotropic hormone induced premature and dystrophic catagen development in anagen HF, whereas inhibitors of MC degranulation retarded normal catagen development. Comparing HF cycling in MC deficient WBB6F1-KitW/KitWv and congenic normal (+/+) mice, catagen development was retarded in the virtual absence of MC. These data support the notion that MC function as hair cycle regulators and are involved in the control of HF regression. The mouse model employed here offers an excellent tool for dissecting the physiologic role of MC as "central switchboards of tissue remodeling" in developmentally regulated systems, specifically in organ involution processes. PMID- 9354768 TI - Membrane CD22 defines circulating myeloma-related cells as mature or later B cells. AB - Circulating clonally related earlier cells are present in multiple myeloma (MM) and may be involved in the dissemination of this disease, its recurrence, and chemoresistance. The nature, stage of differentiation, and size of this cell population remain uncertain. Unlike other B-cell markers, CD22 membrane expression is limited to the late differentiation stages comprised between mature B cells (CD22+) and plasma cells (PC; CD22-) and may thus be useful in delimiting the maturational state of circulating early myeloma cells. Peripheral blood clone related cells were detected by anti-idiotypic (Id) monoclonal antibodies and found to express CD22 (92% to 95%), monotypic light and heavy chain (100%), and CD38 (45%), whereas bone marrow PC were CD22-negative. CD22 expression was also documented on functional myeloma PC precursors, defined as peripheral blood cells capable of in vitro cytokine-driven monotypic PC differentiation, because up to approximately 70% inhibition of this process was obtained in 10 myeloma patients through the use of biospecific antibodies (BsAb) that deliver the plant toxin saporin to CD22+ cells. As further evidence of CD22 on circulating abnormal cells, it was found that in the only patient analyzed for DNA content, a portion of the peripheral blood CD22+ cells killed were hyperdiploid. Collectively, these findings indicate that most peripheral blood myeloma PC precursors are mature or later B cells presenting membrane CD22. The pattern of CD22 expression suggests the existence in MM of a differentiation process analogous to the normal antigenic response, in which CD22+ B cells migrate to the bone marrow and lose CD22 with PC differentiation. In addition, the sensitivity of myeloma PC precursors to the cytotoxic effects of the anti-CD22 BsAb and the possibility of interfering with their differentiation have potential therapeutic relevance. PMID- 9354769 TI - Expression of matrix metalloprotease-9 in vascular pericytes in human breast cancer. AB - Matrix metalloprotease-9 (MMP-9; 92-kd type IV collagenase, gelatinase B) is regarded as important for degradation of the basement membrane and extracellular matrix during cancer invasion and other tissue-remodeling events. Expression of MMP-9 was analyzed in 22 cases of human ductal breast cancer by immunohistochemistry and in 8 of these cases also by in situ hybridization. For immunohistochemistry we used affinity-purified polyclonal antibodies as well as a MMP-9-specific monoclonal antibody (clone 6-6B). Three different stromal cell types with a positive MMP-9 immunoreaction were identified morphologically: neutrophils and macrophage-like cells in all cases and vascular cells in 16 of 22 cases. Double immunofluorescence with antibodies to CD68 conclusively demonstrated MMP-9 expression in macrophages. To identify the positive vascular cells, we employed antibodies to von Willebrand factor and PAL-E for identification of endothelial cells, high molecular weight melanoma-associated antigen for pericytes, and alpha-smooth muscle actin for vascular smooth muscle cells. Using conventional and confocal double immunofluorescence microscopy, colocalization of MMP-9 was seen with high molecular weight melanoma-associated antigen, the pericyte marker, whereas little or no coexpression was seen with alpha-smooth muscle actin. Virtually no coexpression was seen with the endothelial cell markers PAL-E and von Willebrand factor. In situ hybridization showed that MMP-9 mRNA colocalized with MMP-9 immunoreactivity in macrophages and vascular structures, whereas no MMP-9 mRNA was detected in neutrophils. No MMP-9 immunostaining or in situ hybridization signal was detected in cancer cells in any of the cases. Based on these results, it is concluded that MMP-9 in human breast cancer is located in tumor-infiltrating stromal cells, including neutrophils, macrophages, and vascular pericytes, and that the latter two cell types also produce this metalloprotease. We suggest that the MMP-9 produced in pericytes may play a role in extracellular matrix degradation during tumor angiogenesis. PMID- 9354770 TI - Astrocyte-derived cytokines contribute to the metastatic brain specificity of breast cancer cells. AB - The occurrence of breast cancer metastases is preferential to certain organs. Astrocytes may play an important role in the development of brain metastases, as these cells have been shown to respond to extracellular stimuli by producing many cytokines and growth factors that can modulate tumor cell proliferation, growth, and/or metastases. To test this hypothesis, we analyzed the responses of the human breast cancer cell line MDA-MB-435 and its metastatic sublines to astrocyte primary cultures from newborn rat cerebra. Astrocyte purity of the glial cell cultures was demonstrated by glial fibrillary acidic protein and rat neural antigen-2 (Ran-2) immunopositive staining. The 435-Br1 cell line, which was derived from a brain metastases in a nude mouse, showed increased adhesion to astrocytes and enhanced growth in vitro in the presence of media from Con A stimulated astrocytes, relative to the parental MDA-MB-435 and the lung metastasis-derived variant 435-Lung2. Furthermore, the growth-stimulatory effect was partially reversed by anti-IL-6, anti-transforming growth factor beta (anti TGF beta), and anti-IGF-I antibodies, indicating that these metastatic cells use exogenous cytokines as paracrine growth factors. In an attempt to elucidate the role of several biologic-response modifiers produced by astrocytes, we tested the responses of MDA-MB-435 cells to purified cytokines and growth factors. We found that the addition of recombinant human or mouse IL-6 produced a variety of responses in the different 435 metastatic variants. Furthermore, IL-6 receptor (IL-6R) expression was slightly increased in the 435-Br1 cells, and exogenous IL 6 rescued 435-Br1 cells from apoptosis in serum-depleted cultures. No apoptotic protective effect was observed in either MDA-MB-435 parental cells or 435-Lung2 cells. Thus, responses to exogenous IL-6 might determine the differences among these metastatic variants by extending cell survival of selected subpopulations, giving them the opportunity to respond to growth factors or other favorable conditions that might be present. These results suggest that cytokines produced by glial cells in vivo may contribute, in a paracrine manner, to the development of brain metastases by breast cancer cells. PMID- 9354771 TI - Wild-type p53 induces apoptosis in Hep3B through up-regulation of bax expression. AB - We demonstrated that introduction and expression of wild-type p53 gene in the human hepatocellular carcinoma cell line, Hep3B, resulted in up-regulation of both p21WAF1/CIP1 and bax gene expression and apoptosis. This cell line contains integrated hepatitis B virus sequences and lacks the expression of both p53 and retinoblastoma tumor suppressor genes because of deletions. Our results suggest that whereas an increased level of bax expression mediates apoptosis, an increased level of p21WAF1/CIP1 expression does not induce arrest of cell growth, presumably because of the deletion of the retinoblastoma gene. This study also confirms reported observations that p53 is a tumor suppressor gene, which induces apoptosis in malignant cells that lack normal p53 activity because of mutation, deletion, or inactivation of the gene by the presence of oncogenic viral proteins. PMID- 9354773 TI - Fibroblast growth factor receptor-1 is a critical component for endometrial remodeling: localization and expression of basic fibroblast growth factor and FGF R1 in human endometrium during the menstrual cycle and decreased FGF-R1 expression in menorrhagia. AB - Angiogenic growth factors play a critical role in the cyclic growth and vascularization of normal endometrium. Herein, we report the expression and localization of both basic fibroblast growth factor (FGF-2) and its receptor (FGF R1; flg) in human endometrium and demonstrate the markedly decreased FGF-R1 levels in menorrhagia. In situ hybridization using [35S]-labeled riboprobe demonstrated distinct autoradiographic signals for FGF-2 mRNA in glandular epithelial and stromal cells in endometrium throughout the menstrual cycle, with the strongest hybridization signal in stromal cells of the proliferative endometrium relative to that of the secretory endometrium. Moreover, RNAse protection assay revealed that the mRNA encoding FGF-2 and FGF-R1 was significantly higher in proliferative than in secretory endometrium (p < 0.05, p < 0.01). Immunohistochemistry using anti-flg antibody showed that the intensity of FGF-R1 staining was markedly diminished in the stromal cells of secretory endometrium, which corresponded with the reduced FGF-2 mRNA expression. In contrast, the endometrial glandular epithelial cells showed intense localization of FGF-R1 protein throughout the menstrual cycle, which paralleled FGF-2 mRNA expression. Colocalization of FGF-2 and FGF-R1 in stroma and stimulation of DNA synthesis and phospholipase C activation by FGF-2 in these cells demonstrates that FGF-2 acts in an autocrine manner in endometrial stroma. Western immunoblotting showed that FGF-R1 immunoprotein was markedly reduced or absent in women with menorrhagia throughout the cycle relative to that of normal cycling women, suggesting that FGF-R1 is critical for endometrial "maturation" and regeneration of the normal endometrium following menstruation. PMID- 9354772 TI - Cellular distribution of phosphorothioate oligodeoxynucleotides in normal rodent tissues. AB - The distribution of intravenously injected phosphorothioate oligodeoxynucleotides (P = S ODN) was studied in vivo in rodent tissues using three histologic methods: immunohistochemistry with a monoclonal antibody that recognizes P = S ODN ISIS 2105; direct fluorescence microscopy of P = S ODN ISIS 2105 conjugated to rhodamine; and autoradiography of 14C-labeled P = S ODN ISIS 2302. All three methods gave the same pattern of oligonucleotide distribution, and the intensity of the histologic signal agreed with previously published pharmacokinetic data on the relative concentration of P = S ODN in different organs. Proximal tubule cells in the kidney and Kupffer and endothelial cells in the liver were among the most heavily labeled with P = S ODN at all doses and time-points. Connective tissues proper, such as the lamina propria and submucosa of the intestine and the dermis and subcutaneous layer of the skin, were also labeled, whereas the P = S ODN signal was weak or negative in epithelial and muscle cells in the skin and intestine. At 2 hours postinjection, P = S ODN were clearly detectable in the extracellular matrix in loose and dense connective tissues, although by 24 hours, the label was predominantly intracellular. Large, nucleated cells in red marrow, and the connective tissues around bone and skeletal muscle cells and lining the knee joint, were positive for oligonucleotide, whereas P = S ODN were not detected in erythrocytes, cartilage, compact bone, and skeletal muscle. In spleen, white pulp was negative for P = S ODN, whereas cells surrounding the sinusoids and nucleated cells in the red pulp were strongly positive for P = S ODN. Our results provide specific information on the tissue and cellular localization of P = S ODN within organs in vivo. The data presented will be used as a reference for studies of P = S ODN distribution in diseased tissues and the distribution of modified oligonucleotides. Furthermore, because our results indicate which cell types are likely to be affected by antisense oligonucleotides, they can be used to guide future in vivo applications of the technology. PMID- 9354774 TI - No stranger to controversy. PMID- 9354775 TI - Making sense of glucose sensing. PMID- 9354776 TI - Freeze! PMID- 9354777 TI - Left, right ... which way to turn? PMID- 9354778 TI - Advances fuel Alzheimer's conundrum. PMID- 9354779 TI - A tale of mice without 'tails'. PMID- 9354780 TI - Strike three for GLI3. PMID- 9354781 TI - Lack of apolipoprotein E dramatically reduces amyloid beta-peptide deposition. PMID- 9354782 TI - PEX12 encodes an integral membrane protein of peroxisomes. PMID- 9354783 TI - KCNE1 mutations cause jervell and Lange-Nielsen syndrome. PMID- 9354784 TI - Autosomal dominant non-syndromic deafness caused by a mutation in the myosin VIIA gene. PMID- 9354785 TI - Mutation in GLI3 in postaxial polydactyly type A. PMID- 9354786 TI - Germline mutation of MSH6 as the cause of hereditary nonpolyposis colorectal cancer. PMID- 9354787 TI - Overexpression of Agrt leads to obesity in transgenic mice. PMID- 9354788 TI - The pre-implantation ontogeny of the H19 methylation imprint. PMID- 9354789 TI - Communal discourse as a supplement to informed consent for genetic research. AB - Genetic technologies present unique problems for the practice of informed consent. They provide information that may affect a study participant's family or kindred, which may be identifiable as an ethnic or locally isolated population. That information may be used to construct adverse perceptions of such identifiable populations, including non-participants who may not have been informed of or consented to the analyses. To address collective implications of genetic research, we describe a process that can supplement individual consent. Our approach engages pre-existing social units in discourses about proposed research. Communal discourses can influence individuals' decisions to participate in research studies. PMID- 9354790 TI - Theoretical and empirical issues for marker-assisted breeding of congenic mouse strains. AB - Congenic breeding strategies are becoming increasingly important as a greater number of complex trait linkages are identified. Traditionally, the development of a congenic strain has been a time-consuming endeavour, requiring ten generations of backcrosses. The recent advent of a dense molecular genetic map of the mouse permits methods that can reduce the time needed for congenic-strain production by 18-24 months. We present a theoretical evaluation of marker assisted congenic production and provide the empirical data that support it. We present this 'speed congenic' method in a user-friendly manner to encourage other investigators to pursue this or similar methods of congenic production. PMID- 9354791 TI - Opitz G/BBB syndrome, a defect of midline development, is due to mutations in a new RING finger gene on Xp22. AB - Opitz syndrome (OS) is an inherited disorder characterized by midline defects including hypertelorism, hypospadias, lip-palate-laryngotracheal clefts and imperforate anus. We have identified a new gene on Xp22, MID1 (Midline 1), which is disrupted in an OS patient carrying an X-chromosome inversion and is also mutated in several OS families. MID1 encodes a member of the B-box family of proteins, which contain protein-protein interaction domains, including a RING finger, and are implicated in fundamental processes such as body axis patterning and control of cell proliferation. The association of MID1 with OS suggests an important role for this gene in midline development. PMID- 9354792 TI - A novel gene involved in zinc transport is deficient in the lethal milk mouse. AB - We have identified the gene responsible for the inherited zinc deficiency in the lethal milk (lm) mouse. The gene, here designated Znt4, encodes a 430-amino-acid protein that is homologous to two proteins, ZnT2 and ZnT3, responsible for sequestration of zinc into endosomal/lysosomal compartments and synaptic vesicles, respectively. We show that the Znt4 gene confers zinc resistance to a zinc-sensitive yeast strain and that it is abundantly expressed in the mammary epithelia and brain. The lethal milk mutant has a nonsense mutation at arginine codon 297 in the Znt4 gene. PMID- 9354793 TI - Trinucleotide repeats affect DNA replication in vivo. AB - (CGG)n.(CCG)n and (CTG)n.(CAG)n repeats of varying length were cloned into a bacterial plasmid, and the progression of the replication fork through these repeats was followed using electrophoretic analysis of replication intermediates. We observed stalling of the replication fork within repeated DNAs and found that this effect depends on repeat length, repeat orientation relative to the replication origin and the status of protein synthesis in a cell. Interruptions within repeated DNAs, similar to those observed in human genes, abolished the replication blockage. Our results suggest that the formation of unusual DNA structures by trinucleotide repeats in the lagging-strand template may account for the observed replication blockage and have relevance to repeat expansion in humans. PMID- 9354794 TI - X-linked situs abnormalities result from mutations in ZIC3. AB - Vertebrates position unpaired organs of the chest and abdomen asymmetrically along the left-right (LR) body axis. Each structure comes to lie non-randomly with respect to the midline in an overall position designated situs solitus, exemplified in humans by placement of the heart, stomach and spleen consistently to the left. Aberrant LR axis development can lead to randomization of individual organ position (situs ambiguus) or to mirror-image reversal of all lateralized structures (situs inversus). Previously we mapped a locus for situs abnormalities in humans, HTX1, to Xq26.2 by linkage analysis in a single family (LR1) and by detection of a deletion in an unrelated situs ambiguus male (Family LR2; refs 2,3). From this chromosomal region we have positionally cloned ZIC3, a gene encoding a putative zinc-finger transcription factor. One frameshift, two missense and two nonsense mutations have been identified in familial and sporadic situs ambiguus. The frameshift allele is also associated with situs inversus among some heterozygous females, suggesting that ZIC3 functions in the earliest stages of LR-axis formation. ZIC3, which has not been previously implicated in vertebrate LR-axis development, is the first gene unequivocally associated with human situs abnormalities. PMID- 9354795 TI - The EWS-WT1 translocation product induces PDGFA in desmoplastic small round-cell tumour. AB - Chromosomal translocations resulting in chimaeric transcription factors underlie specific malignancies, but few authentic target genes regulated by these fusion proteins have been identified. Desmoplastic small round-cell tumour (DSRT) is a multiphenotypic primitive tumour characterized by massive reactive fibrosis surrounding nests of tumour cells. The t(11;22)(p13;q12) chromosomal translocation that defines DSRT produces a chimaeric protein containing the potential transactivation domain of the Ewing-sarcoma protein (EWS) fused to zinc fingers 2-4 of the Wilms tumour suppressor and transcriptional repressor WT1 (refs 2,3). By analogy with other EWS fusion products, the EWS-WT1 chimaera may encode a transcriptional activator whose target genes overlap with those repressed by WT1 (ref. 4). To characterize its functional properties, we generated osteosarcoma cell lines with tightly regulated inducible expression of EWS-WT1. Expression of EWS-WT1 induced the expression of endogenous platelet derived growth factor-A (PDGFA), a potent secreted mitogen and chemoattractant whose promoter contains the many potential WT1-binding sites. Native PDGFA was not regulated by wild-type WT1, indicating a difference in target gene specificity between this tumour suppressor and its oncogenic derivative. PDGFA was expressed within tumour cells in primary DSRT specimens, but it was absent in Wilms tumours expressing WT1 and Ewing sarcomas with an EWS-Fli translocation. We conclude that the oncogenic fusion of EWS to WT1 in DSRT results in the induction of PDGFA, a potent fibroblast growth factor that contributes to the characteristic reactive fibrosis associated with this unique tumour. PMID- 9354796 TI - Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors. AB - Successful gene therapy approaches will require efficient gene delivery and sustained expression of the transgene in recipients. A variety of methods, ranging from direct DNA delivery to infection with recombinant viruses containing foreign genes, have been developed, but they all have some major limitations that restrict their utility. We have described a human lentiviral (HIV)-based vector that can transduce non-dividing cells in vitro and deliver genes in vivo. With this vector, expression of transgenes in the brain has been detected for more than six months--the longest period tested so far. Because lentiviral vectors are pseudotyped with vesicular stomatitis virus G glycoprotein (VSVG; ref. 8), they can transduce a broad range of tissues and cell types. We now describe the ability of lentiviral vectors to introduce genes directly into liver and muscle. Sustained expression of green fluorescent protein (GFP), used as a surrogate for therapeutic protein, can be observed for more than 22 weeks in the liver. Similar long-term expression (more than eight weeks) was observed in transduced muscle. In contrast, little or no GFP could be detected in liver or muscle transduced with the Moloney murine leukaemia virus (M-MLV), a prototypic retroviral based vector. At a minimum, 3-4% of the total liver tissue was transduced by a single injection of 1-3 x 10(7) infectious units (I.U.) of recombinant HIV vector. Furthermore, no inflammation of recruitment of lymphocytes could be detected at the site of injection. Animals previously transduced with a lentiviral vector can be efficiently re-infected with lentiviral vectors. Additionally, we show that the requirement for lentiviral accessory proteins to establish efficient transduction in vivo is tissue dependent. PMID- 9354797 TI - Absence of integrin alpha 7 causes a novel form of muscular dystrophy. AB - Integrin alpha 7 beta 1 is a specific cellular receptor for the basement membrane protein laminin-1 (refs 1,2), as well as for the laminin isoforms -2 and -4 (ref. 3). The alpha 7 subunit is expressed mainly in skeletal and cardiac muscle and has been suggested to be involved in differentiation and migration processes during myogenesis. Three cytoplasmic and two extracellular splice variants that have been described are developmentally regulated and expressed in different sites in the muscle. In adult muscle, the alpha 7A and alpha 7B subunits are concentrated in myotendinous junctions but can also be detected in neuromuscular junctions and along the sarcolemmal membrane. To study the potential involvement of alpha 7 integrin, during myogenesis and its role in muscle integrity and function, we generated a null allele of the alpha 7 gene (Itga7) in the germline of mice by homologous recombination in embryonic stem (ES) cells. Surprisingly, mice homozygous for the mutation are viable and fertile, indicating that the alpha 7 beta 1 integrin is not essential for myogenesis. However, histological analysis of skeletal muscle revealed typical symptoms of a progressive muscular dystrophy starting soon after birth, but with a distinct variability in different muscle types. The observed histopathological changes strongly indicate an impairment of function of the myotendinous junctions. These findings demonstrate that alpha 7 beta 1 integrin represents an indispensable linkage between the muscle fibre and the extracellular matrix that is independent of the dystrophin dystroglycan complex-mediated interaction of the cytoskeleton with the muscle basement membrane. PMID- 9354798 TI - Mutations in GLUT2, the gene for the liver-type glucose transporter, in patients with Fanconi-Bickel syndrome. AB - Fanconi-Bickel syndrome (FBS) is a rare autosomal-recessive inborn error of metabolism characterized by hepatorenal glycogen accumulation, Fanconi nephropathy and impaired utilization of glucose and galactose. To date, no underlying enzymatic defect in carbohydrate metabolism has been identified. Therefore, and because of the impairment of both glucose and galactose metabolism, a primary defect of monosaccharide transport across membranes has been suggested. Here we report mutations in the gene encoding the facilitative glucose transporter 2 (GLUT2) in three FBS families, including the original patient described in 1949 by Fanconi and Bickel. Homozygous mutations were found in affected individuals, whereas all parents tested were heterozygous for the respective mutation. Because all detected mutations (delta T446-449, C1251T and C1405T) predict truncated translation products that cannot be expected to have functional monosaccharide transport activity, GLUT2 mutations are probably the cause of FBS. PMID- 9354799 TI - Early diabetes and abnormal postnatal pancreatic islet development in mice lacking Glut-2. AB - Glut-2 is a low-affinity transporter present in the plasma membrane of pancreatic beta-cells, hepatocytes and intestine and kidney absorptive epithelial cells of mice. In beta-cells, Glut-2 has been proposed to be active in the control of glucose-stimulated insulin secretion (GSIS; ref. 2), and its expression is strongly reduced in glucose-unresponsive islets from different animal models of diabetes. However, recent investigations have yielded conflicting data on the possible role of Glut-2 in GSIS. Whereas some reports have supported a specific role for Glut-2 (refs 5,6), others have suggested that GSIS could proceed normally even in the presence of low or almost undetectable levels of this transporter. Here we show that homozygous, but not heterozygous, mice deficient in Glut-2 are hyperglycaemic and relatively hypo-insulinaemic and have elevated plasma levels of glucagon, free fatty acids and beta-hydroxybutyrate. In vivo, their glucose tolerance is abnormal. In vitro, beta-cells display loss of control of insulin gene expression by glucose and impaired GSIS with a loss of first phase but preserved second phase of secretion, while the secretory response to non-glucidic nutrients or to D-glyceraldehyde is normal. This is accompanied by alterations in the postnatal development of pancreatic islets, evidenced by an inversion of the alpha- to beta-cell ratio. Glut-2 is thus required to maintain normal glucose homeostasis and normal function and development of the endocrine pancreas. Its absence leads to symptoms characteristic of non-insulin-dependent diabetes mellitus. PMID- 9354800 TI - Quantitative trait locus analysis of contextual fear conditioning in mice. AB - Family, twin and adoption studies provide evidence for a substantial genetic component underlying individual differences in general intelligence, specific cognitive abilities and susceptibility to psychopathologies related to fear inducing events. Contextual fear conditioning, which is highly conserved across species, can serve as a model for elucidating genes that regulate individual differences in learning and emotion. In fear conditioning, an initially neutral stimulus, such as a tone or a particular environment (context), elicits a fear response after it has been paired with an aversive stimulus, such as shock. Two neural circuits have been implicated in fear conditioning. The fear component is regulated by amygdaloid pathways, while the contextual component is, at least in part, dependent on the hippocampus. C57BL/6J (B6) and DBA/2J (D2) mice differ in several types of complex learning. including contextual fear conditioning. A quantitative trait locus (QTL) analysis of contextual fear conditioning was performed in a B6/D2 F2 intercross population. Two QTLs for contextual conditioning (lod score > 4.3) were identified on chromosomes 10 and 16. QTLs for conditioning to the auditory cue (lod score > 4.3) were localized to chromosomes 1 and 10. Suggestive QTLs (lod score = 2.8-4.1) for contextual conditioning were detected on chromosomes 1, 2 and 3. PMID- 9354801 TI - Quantitative trait loci analysis affecting contextual conditioning in mice. AB - In an extensive backcross of mice between C3H/HeJ (C3H) and C57BL/6J (B6), we sought to map genes that influence learning and memory as measured by performance in a contextual fear-conditioning paradigm. Our results indicate that there are several genetic regions that have a strong influence on performance in this paradigm. The strongest influences map to the proximal and distal ends of chromosome 1 (lod scores of 5.14 and 4.76, respectively). Other chromosomal regions (chromosomes 3, 7, 8, 9 and 18) were also identified as candidates for regions containing genes influencing contextual fear conditioning, with lod scores ranging from 1.8 to 2.7. PMID- 9354802 TI - Mutations in the hminK gene cause long QT syndrome and suppress IKs function. AB - Ion-channel beta-subunits are ancillary proteins that co-assemble with alpha subunits to modulate the gating kinetics and enhance stability of multimeric channel complexes. Despite their functional importance, dysfunction of potassium channel beta-subunits has not been associated with disease. Recent physiological studies suggest that KCNE1 encodes beta-subunits (hminK) that co-assemble with KvLQT1 alpha-subunits to form the slowly activating delayed rectifier K+ (IKs) channel. Because KVLQT1 mutations cause arrhythmia susceptibility in the long QT syndrome (LQT), we hypothesized that mutations in KCNE1 also cause this disorder. Here, we define KCNE1 missense mutations in affected members of two LQT families. Both mutations (S74L, D76N) reduced IKs by shifting the voltage dependence of activation and accelerating channel deactivation. D76N hminK also had a strong dominant-negative effect. The functional consequences of these mutations would be delayed cardiac repolarization and an increased risk of arrhythmia. This is the first description of KCNE1 as an LQT gene and confirms that hminK is an integral protein of the IKs channel. PMID- 9354803 TI - BRCA1 genomic deletions are major founder mutations in Dutch breast cancer patients. AB - To date, more than 300 distinct small deletions, insertions and point mutations, mostly leading to premature termination of translation, have been reported in the breast/ovarian-cancer susceptibility gene BRCA1. The elevated frequencies of some mutations in certain ethnic subpopulations are caused by founder effects, rather than by mutation hotspots. Here we report that the currently available mutation spectrum of BRCA1 has been biased by PCR-based mutation-screening methods, such as SSCP, the protein truncation test (PTT) and direct sequencing, using genomic DNA as template. Three large genomic deletions that are not detected by these approaches comprise 36% of all BRCA1 mutations found in Dutch breast-cancer families to date. A 510-bp Alu-mediated deletion comprising exon 22 was found in 8 of 170 breast-cancer families recruited for research purposes and in 6 of 49 probands referred to the Amsterdam Family Cancer Clinic for genetic counselling. In addition, a 3,835-bp Alu-mediated deletion encompassing exon 13 was detected in 4 of 170 research families, while an deletion of approximately 14 kb was detected in a single family [corrected]. Haplotype analyses indicated that each recurrent deletion had a single common ancestor. PMID- 9354804 TI - Disruption of the mouse L1 gene leads to malformations of the nervous system. AB - The adhesion molecule L1 is a member of the immunoglobulin superfamily. L1 is involved in various recognition processes in the CNS and PNS, and binding to L1 can activate signal transduction pathways. Mutations in the human L1 gene are associated with a variable phenotype, including mental retardation and anomalous development of the nervous system, referred to as 'CRASH' (corpus callosum hypoplasia, retardation, adducted thumbs, spastic paraplegia, and hydrocephalus). We generated an animal model of these conditions by gene targetting. Mutant mice were smaller than wild-type and were less sensitive to touch and pain, and their hind-legs appeared weak and uncoordinated. The size of the corticospinal tract was reduced and, depending on genetic background, the lateral ventricles were often enlarged. Non-myelinating Schwann cells formed processes not associated with axons and showed reduced association with axons. In vitro, neurite outgrowth on an L1 substrate and fasciculation were impaired. The mutant mouse described here will help to elucidate the functions of L1 in the nervous system and how these depend on genetic influences. PMID- 9354805 TI - Insulin VNTR allele-specific effect in type 1 diabetes depends on identity of untransmitted paternal allele. The IMDIAB Group. AB - The IDDM2 type 1 diabetes susceptibility locus was mapped to and identified as allelic variation at the insulin gene (INS) VNTR regulatory polymorphism. In Caucasians, INS VNTR alleles divide into two discrete size classes. Class I alleles (26 to 63 repeats) predispose in a recessive way to type 1 diabetes, while class III alleles (140 to more than 200 repeats) are dominantly protective. The protective effect may be explained by higher levels of class III VNTR associated INS mRNA in thymus such that elevated levels of preproinsulin protein enhance immune tolerance to preproinsulin, a key autoantigen in type 1 diabetes pathogenesis. The mode of action of IDDM2 is complicated, however, by parent-of origin effects and possible allelic heterogeneity within the two defined allele classes. We have now analysed transmission of specific VNTR alleles in 1,316 families and demonstrate that a particular class I allele does not predispose to disease when paternally inherited, suggestive of polymorphic imprinting. But this paternal effect is observed only when the father's untransmitted allele is a class III. This allelic interaction is reminiscent of epigenetic phenomena observed in plants (for example, paramutation; ref. 17) and in yeast (for example, trans-inactivation; ref. 18). If untransmitted chromosomes can have functional effects on the biological properties of transmitted chromosomes, the implications for human genetics and disease are potentially considerable. PMID- 9354806 TI - A promoter mutation in the XIST gene in two unrelated families with skewed X chromosome inactivation. AB - X-chromosome inactivation is the process by which a cell recognizes the presence of two copies of an X chromosome early in the development of XX embryos and chooses one to be active and one to be inactive. Although it is commonly believed that the initiation of X inactivation is random, with an equal probability (50:50) that either X chromosome will be the inactive X in a given cell, significant variation in the proportion of cells with either X inactive is observed both in mice heterozygous for alleles at the Xce locus and among normal human females in the population. Families in which multiple females demonstrate extremely skewed inactivation patterns that are otherwise quite rare in the general population are thought to reflect possible genetic influences on the X inactivation process. Here we report a rare cytosine to guanine mutation in the XIST minimal promoter that underlies both epigenetic and functional differences between the two X chromosomes in nine females from two unrelated families. All females demonstrate preferential inactivation of the X chromosome carrying the mutation, suggesting that there is an association between alterations in the regulation of XIST expression and X-chromosome inactivation. PMID- 9354807 TI - The human necdin gene, NDN, is maternally imprinted and located in the Prader Willi syndrome chromosomal region. AB - Prader-Willi syndrome (PWS) is a neurogenetic disorder that results from the absence of a normal paternal contribution to the 15q11-13 region. The clinical manifestations of PWS are a transient severe hypotonia in the newborn period, with mental retardation, hypogonadism and obesity observed later in development. Five transcripts with exclusive expression from the paternal allele have been isolated, but none of these has been shown to be involved in PWS. In this study, we report the isolation and characterization of NDN, a new human imprinted gene. NDN is exclusively expressed from the paternal allele in the tissues analysed and is located in the PWS region. It encodes a putative protein homologous to the mouse brain-specific NECDIN protein, NDN; as in mouse, expression in brain is restricted to post-mitotic neurons. NDN displays several characteristics of an imprinted locus, including allelic DNA methylation and asynchronous DNA replication. A complete lack of NDN expression in PWS brain and fibroblasts indicates that the gene is expressed exclusively from the paternal allele in these tissues and suggests a possible role of this new gene in PWS. PMID- 9354808 TI - Store depletion and calcium influx. AB - Calcium influx in nonexcitable cells regulates such diverse processes as exocytosis, contraction, enzyme control, gene regulation, cell proliferation, and apoptosis. The dominant Ca2+ entry pathway in these cells is the store-operated one, in which Ca2+ entry is governed by the Ca2+ content of the agonist-sensitive intracellular Ca2+ stores. Only recently has a Ca2+ current been described that is activated by store depletion. The properties of this new current, called Ca2+ release-activated Ca2+ current (ICRAC), have been investigated in detail using the patch-clamp technique. Despite intense research, the nature of the signal that couples Ca2+ store content to the Ca2+ channels in the plasma membrane has remained elusive. Although ICRAC appears to be the most effective and widespread influx pathway, other store-operated currents have also been observed. Although the Ca2+ release-activated Ca2+ channel has not yet been cloned, evidence continues to accumulate that the Drosophila trp gene might encode a store operated Ca2+ channel. In this review, we describe the historical development of the field of Ca2+ signaling and the discovery of store-operated Ca2+ currents. We focus on the electrophysiological properties of the prototype store-operated current ICRAC, discuss the regulatory mechanisms that control it, and finally consider recent advances toward the identification of molecular mechanisms involved in this ubiquitous and important Ca2+ entry pathway. PMID- 9354809 TI - Immunomodulatory functions of surfactant. AB - The possibility that the lipoprotein complex of lung surfactant functions in pulmonary host defense as well as lowering surface tension at the air-liquid interface has been the subject of renewed interest in light of the finding that surfactant proteins A and D (SP-A and SP-D) are members of a family of proteins known as collectins. The collectins, so named because they have in common an NH2 terminal collagen-like domain and a COOH-terminal lectin (carbohydrate binding) domain, are found in both lung and serum and participate in "innate" immunity, acting before induction of an antibody-mediated response. In vitro, many of the collectins stimulate phagocytosis, chemotaxis, and production of reactive oxygen and regulate cytokine release by immune cells. It has been known for several years that surfactant lipids suppress a variety of immune cell functions, most notably lymphocyte proliferation, which, conversely, is augmented by SP-A. Thus surfactant lipids and proteins may be counterregulatory, and changes in lipid-to protein ratios may be important in regulating the immune status of the lung. That these ratios change in disease states is clear, but it is not known whether the alterations are a cause or an effect. Important future studies with mice in which the SP-A and SP-D genes have been ablated will help clarify the role of surfactant in immune function. PMID- 9354810 TI - Hormonal regulation in insects: facts, gaps, and future directions. AB - There are two main classes of hormones in insects: 1) the true hormones produced by epithelial glands and belonging to the ecdysteroids or juvenile hormones and 2) the neuropeptide hormones produced by neurosecretory cells. Members of these classes regulate physiological, developmental, and behavioral events in insects. Detailed accounts are given on isolation, identification, structure-activity relationships, mode of action, biological function, biosynthesis, inactivation, metabolism, and feedback for hormones involved in 1) metabolic regulation such as the adipokinetic/hypertrehalosemic peptides and the diuretic and antidiuretic peptides; 2) stimulation or inhibition of muscle activity such as the myotropic peptides; 3) control of reproduction, growth, and development such as allatotropins, allatostatins, juvenile hormones, ecdysteroids, folliculostimulins and folliculostatins, ecdysis-triggering and eclosion hormones, pheromone biosynthesis activating neuropeptides, and diapause hormones; and 4) regulation of tanning and of color change. Because of the improvements in techniques for isolation and structure elucidation, there has been rapid progress in our knowledge of the chemistry of certain neuropeptide families. With the employment of molecular biological techniques, the genes of some neuropeptides have been successfully characterized. There are, however, areas that are still quite underdeveloped. These are, for example, 1) receptor studies, which are still in their infancy; 2) the hormonal status of certain sequenced peptides is not clarified; and 3) functional studies are lacking even for established hormones. The authors plead for a concerted effort to continue research in this field, which will also advance our knowledge into the use of insect hormones as safer and species-specific molecules for insect pest management. PMID- 9354812 TI - Cellular actions of beta-amyloid precursor protein and its soluble and fibrillogenic derivatives. AB - beta-Amyloid precursor protein (beta-APP), the source of the fibrillogenic amyloid beta-peptide (A beta) that accumulates in the brain of victims of Alzheimer's disease, is a multifunctional protein that is widely expressed in the nervous system. beta-Amyloid precursor protein is axonally transported and accumulates in presynaptic terminals and growth cones. A secreted form of beta APP (sAPP alpha) is released from neurons in response to electrical activity and may function in modulation of neuronal excitability, synaptic plasticity, neurite outgrowth, synaptogenesis, and cell survival. A signaling pathway involving guanosine 3',5'-cyclic monophosphate is activated by sAPP alpha and modulates the activities of potassium channels, N-methyl-D-aspartate receptors, and the transcription factor NF kappa B. Additional functions of beta-APP may include modulation of cell adhesion and regulation of proliferation of nonneuronal cells. Alternative enzymatic processing of beta-APP liberates A beta, which has a propensity to form amyloid fibrils; A beta can damage and kill neurons and increase their vulnerability to excitotoxicity. The mechanism involves generation of oxyradicals and impairment of membrane transport systems (e.g., ion-motive ATPases and glutamate and glucose transporters). Genetic mutations or age-related metabolic changes may promote neuronal degeneration in Alzheimer's disease by increasing production of A beta and/or decreasing levels of neuroprotective sAPP alpha. PMID- 9354811 TI - Mast cells. AB - Mast cells are found resident in tissues throughout the body, particularly in association with structures such as blood vessels and nerves, and in proximity to surfaces that interface the external environment. Mast cells are bone marrow derived and particularly depend upon stem cell factor for their survival. Mast cells express a variety of phenotypic features within tissues as determined by the local environment. Withdrawal of required growth factors results in mast cell apoptosis. Mast cells appear to be highly engineered cells with multiple critical biological functions. They may be activated by a number of stimuli that are both Fc epsilon RI dependent and Fc epsilon RI independent. Activation through various receptors leads to distinct signaling pathways. After activation, mast cells may immediately extrude granule-associated mediators and generate lipid-derived substances that induce immediate allergic inflammation. Mast cell activation may also be followed by the synthesis of chemokines and cytokines. Cytokine and chemokine secretion, which occurs hours later, may contribute to chronic inflammation. Biological functions of mast cells appear to include a role in innate immunity, involvement in host defense mechanisms against parasitic infestations, immunomodulation of the immune system, and tissue repair and angiogenesis. PMID- 9354813 TI - Mechanisms of calcium signaling by cyclic ADP-ribose and NAADP. AB - Cells possess various mechanisms for transducing external signals to intracellular responses. The discovery of inositol 1,4,5-trisphosphate (IP3) as a messenger for mobilizing internal Ca2+ stores has centralized Ca2+ mobilization among signaling mechanisms. Results reviewed in this article establish that, in addition to IP3, the internal Ca2+ stores can be mobilized by at least two other molecules, cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP), via totally independent mechanisms. Cyclic ADP-ribose is a newly discovered cyclic nucleotide derived from NAD, but, unlike adenosine 3',5' cyclic monophosphate, its main signaling function is modulation of Ca(2+)-induced Ca2+ release, a major mechanism of Ca2+ mobilization in addition to the IP3 pathway. Evidence shows that cADPR may in fact be responsible for mediating the Ca(2+)-mobilizing activity of the gaseous messenger nitric oxide. Cells responsive to cADPR are widespread and include species from plant to mammal, indicating the generality of cADPR as a signaling molecule. In addition to cADPR, NAADP, a metabolite of NADP, can also mobilize Ca2+ stores. The release mechanism and the stores on which NAADP acts are distinct from cADPR and IP3. Nicotinic acid adenine dinucleotide phosphate may play a role in generating Ca2+ oscillations, since liberation of NAADP in live cells by photolyzing its caged analog produces long lasting Ca2+ oscillations. These two new Ca2+ agonists are intimately related, since the same metabolic enzymes can, under appropriate conditions, synthesize either one, suggesting a unified mechanism may regulate both pathways. Elucidation of these two new Ca2+ mobilization pathways is likely to have an important impact on our understanding of cellular signaling mechanisms. PMID- 9354815 TI - Apheresis: man versus machine. PMID- 9354814 TI - ATP-sensitive and inwardly rectifying potassium channels in smooth muscle. AB - The properties and roles of ATP-sensitive (KATP) and inwardly rectifying (KIR) potassium channels are reviewed. Potassium channels regulate the membrane potential of smooth muscle, which controls calcium entry through voltage dependent calcium channels, and thereby contractility through changes in intracellular calcium. The KATP channel is likely to be composed of members of the inward rectifier channel gene family (Kir6) and sulfonylurea receptor proteins. The KIR channels do not appear to be as widely distributed as KATP channels in smooth muscle and may provide a mechanism by which changes in extracellular K+ can alter smooth muscle membrane potential, and thereby arterial diameter. The KATP channels contribute to the resting membrane conductance of some types of smooth muscle and can open under situations of metabolic compromise. The KATP channels are targets of a wide variety of vasodilators and constrictors, which act, respectively, through adenosine 3',5'-cyclic monophosphate/protein kinase A and protein kinase C. The KATP channels are also activated by a number of synthetic vasodilators (e.g., diazoxide and pinacidil) and are inhibited by the oral hypoglycemic sulfonylurea drugs (e.g., glibenclamide). Together, KATP and KIR channels are important regulators of smooth muscle function and represent important therapeutic targets. PMID- 9354816 TI - Potential increased risk of virus transmission due to exclusion of older donors because of concern over Creutzfeldt-Jakob disease. The National Heart, Lung, and Blood Institute Retrovirus Epidemiology Donor Study. AB - BACKGROUND: Concern over the theoretical possibility of disease transmission via blood from donors who develop Creutzfeldt-Jakob disease has led to proposals to exclude older individuals from donating plasma for further manufacture into pooled plasma donations. The impact of extending this age-deferral policy to blood donors was examined with respect to the risk for known transmissible viruses. STUDY DESIGN AND METHODS: Demographic characteristics and confirmed prevalence rates (/10(5) first-time donations) and incidence rates (/10(5) person years for repeat donors) for viral markers were compared for donors < 50 years old (n = 1,259,805 [85%]) and > or = 50 years old (n = 219,856 [15%]) and for donors < 60 years old (n = 1,409,176 [95%]) and > or = 60 years old (n = 70,485 [5%]). Incidence rates were combined with infectious window-period estimates for each virus, to calculate the risk of virus transmission per 10(6) donations. RESULTS: Unadjusted prevalence rates were significantly greater for younger than for older donor groups for human immunodeficiency virus (HIV), hepatitis B surface antigen (HBsAg), and hepatitis C virus (HCV) (p < or = 0.05). Incidence rates (and transmission risk estimates) for HBsAg were significantly higher in the < 50 donor group than in the > or = 50 group (p < or = 0.05), and those for HIV, human T-lymphotropic virus, and HCV were not significantly higher (p > 0.05). Blanket removal of donors over the age of 50 would potentially lead to the following significant increases in the risk of infected units: HIV, 12 percent; HCV, 21 percent; and hepatitis B virus (HBsAg), 22 percent. CONCLUSION: Removal of donors over the age of 60 would not significantly affect the risk of infected units. Deferral of donors > or = 50 years of age from whole-blood donations for unfounded concerns about Creutzfeldt-Jakob disease could have adverse effects on both blood availability and safety. PMID- 9354817 TI - Value and cost-effectiveness of screening blood donors for antibody to hepatitis B core antigen as a way of detecting window-phase human immunodeficiency virus type 1 infections. The HIV Blood Donor Study Group. AB - BACKGROUND: The value of screening donors for antibody to hepatitis B core antigen (anti-HBc) for the prevention of posttransfusion hepatitis has declined markedly. However, anti-HBc screening may still be useful as a surrogate marker for the window period (WP) of human immunodeficiency virus type 1 (HIV-1) infection. STUDY DESIGN AND METHODS: First, the relationship between anti-HBc reactivity and HIV-1 WP infections was examined among 225 donors who had seroconverted to anti-HIV-1 positivity between 1987 and 1990. In addition, data from 1654 HIV-1 seropositive donors were analyzed to characterize the relationship among anti-HBc reactivity, donor demographics, and HIV-1-related risk factors. The yield and cost-effectiveness of anti-HBc for HIV-1 prevention were then projected on the basis of a published decision analysis model. RESULTS: Forty (18%) of 225 HIV-1-seroconverting donors tested anti-HBc-reactive on the donation preceding anti-HIV-1 seroconversion; in contrast, 341 (34%) of 1014 HIV 1-seropositive donors interviewed tested anti-HBc-reactive (chi-square test; p < 0.001). Anti-HBc reactivity was more common among HIV-1-seropositive donors reporting male-to-male sexual contact (169/360, 47%) and injection drug use (44/83, 53%) than among those with heterosexual contacts known to be HIV-1 positive (31/190, 16%) or transfusion exposure (3/21, 14%) or among females with no identified risk factors (21/124, 17%). The estimates of 18 to 34 percent sensitivity for anti-HBc in detecting HIV-1 WP donations and a current rate of 1 in 676,000 HIV-1 WP donations (after p24 antigen screening) suggest that continued use of anti-HBc screening could result in the transfusion of 5 to 12 fewer HIV-1-infected units per year in the United States, which would add 19 to 48 quality-adjusted years of life for the 3.5 million annual transfusion recipients at a cost of $992,020 to $2,345,000 per quality-adjusted life-year saved. CONCLUSION: The low yield and very poor cost-effectiveness of anti-HBc screening indicate that this test is not an effective screening test for HIV-1 WP donations. PMID- 9354818 TI - A sequence-specific polymerase chain reaction assay for mitochondrial DNA polymorphisms in human platelets and white cells. AB - BACKGROUND: Because mitochondria are abundant in white cells and are also present in platelets, polymorphic sequences in mitochondrial DNA (mtDNA) represent a unique target for polymerase chain reaction (PCR)-based detection of donor material. STUDY DESIGN AND METHODS: A PCR assay was developed that uses sequence specific primers (SSP) focused on two continent-specific mtDNA polymorphisms. Results were validated by the use of informative restriction endonucleases. Three commercially available methods to extract mtDNA from white cell-reduced human platelets was compared. In preparation for in vivo studies, in vitro mixing studies designed to mimic transfusion were conducted to investigate the performance of the SSP-PCR assay. RESULTS: The gene sequences of two representative examples of amplicons obtained with the new SSP-PCR matched the sequence expected from the published genetic code. Fifteen individuals were classified as either positive (n = 6) or negative (n = 9) for the Asian polymorphism by the use of published primers known to flank the polymorphic site followed by digestion with appropriate restriction enzymes. Results with SSP-PCR were nearly perfectly concordant with those of restriction enzyme analysis. Although the use of three DNA extraction methods allowed the preparation of mtDNA that was suitable for PCR, large and consistent differences (ranging from 10- to 1000-fold) in endpoint sensitivity were found. In vitro mixing studies reproducibly documented that the SSP-PCR assay could detect as little as 1 percent of donor platelets mixed with recipient blood. CONCLUSION: PCR-SSP can be reliably used to identify human mtDNA polymorphisms. By optimization of the method of mtDNA extraction, the sensitivity of PCR-SSP assay was greatly increased. This assay should prove useful in investigations of allogeneic platelet transfusions without cell labeling. It may also be applied to studies of the donor cell microchimerism that follows transfusion or transplantation. PMID- 9354820 TI - Co-presence of a point mutation and a deletion of exon 3 in the glycophorin C gene and concomitant production of a Gerbich-related antibody. AB - BACKGROUND: Antigens of the human blood group system Gerbich (Ge) are located on sialoglycoproteins glycophorin C (GPC) and glycophorin D (GPD). CASE REPORT: The Ge+ proposita (RW) produced an alloanti-Ge after receiving 2 units of red cells (RBCs) during surgery. Further studies were carried out to characterize the antibody specificity, RBC GPC and/or GPD (GPC/GPD), and the glycophorin C gene (GYPC) from RW and her compatible siblings. RW's serum contained an alloanti-Ge that did not react with RBCs from RW or four of her siblings or with RBCs with Ge negative phenotypes. An eluate of RW's antibody reacted weakly with GPC in Western blotting. RW's RBCs were positive with 20 alloanti-Ge2, 1 autoanti-Ge2, 4 alloanti-Ge3, and 1 alloanti-Ge4. Titrations revealed weak expression of these antigens on her RBCs and those of her compatible siblings as compared with controls. In contrast, titrations of mouse and rat monoclonal antibodies specific for GPC/GPD showed no differences. Western blotting of RBC membranes using GPC/GPD specific monoclonal antibodies showed a broad diffuse band corresponding to GPC.Ge in addition to GPC and GPD. Blotting of membranes from trypsin-treated RBCs from these individuals revealed an increase of 1500 in M(r) of membrane bound tryptic fragment over that in the membranes from typsin-treated RBCs from persons with normal GPC/GPD. In RT-PCR, two products were obtained for RW and her compatible siblings: one had a complete deletion of exon 3 and the other had a base change (A-->T) in nucleotide 173 in exon 3 (confirmed by genomic DNA sequencing of exon 3). This point mutation has resulted in the loss of restriction enzyme Tth111 I-sensitive site in the mutant GYPC. CONCLUSION: The specificity of antibody in RW's serum was serologically anti-Ge2. Two genetic events occurred in exon 3 in GYPC of RW and her compatible siblings. The exon 3 deletion confirmed a Ge:-2,-3,4 haplotype. The abnormal tryptic fragment obtained was due to the (A173-->T) base change in exon 3 that resulted in Asp58-->Val in the deduced amino acid sequence at the membrane boundary. PMID- 9354819 TI - RHD/CE typing by polymerase chain reaction using sequence-specific primers. AB - BACKGROUND: Current DNA-based Rh system typing strategies may detect the two RH genes and their prevalent alleles, but they are known to fail sometimes, when rare RH alleles (e.g., D category phenotypes) are encountered. It is almost impossible to find a single DNA-based method that can accommodate the great heterogeneity within the human Rh system. STUDY DESIGN AND METHODS: An easy-to perform DNA-based method for the detection of the two RH genes and their alleles, including variant RHD alleles, was developed. By the use of one RHD/C-, seven RHD , and four RHCE-specific polymerase chain reactions, all triggered to work at identical thermocycling conditions, the DNA of 77 blood donors carrying weak D and that of 200 random donors with common D phenotype was investigated. In addition, 77 selected samples of ccDee and rare Rh system phenotypes were examined. RESULTS: Among 77 samples of weak D, one Rh33 and six DVI categories were detected, one of which showed new RHD-specific nucleotide patterns. In DFR and CCee samples, novel variant RHD alleles were found. RHD DNA types of 200 random donors were found to be concordant with their D phenotype. For RHE and RHe genotyping, a full correlation with serologic phenotypes was found. Our method for genotyping RHC and RHc failed in some cases, because of an already published RHc allelic variation, which we have called RHc(cyt48). An estimate of the frequency of this RHc(cyt48) allele in a white population was made. CONCLUSION: The presented exon-scanning RHD/CE polymerase chain reaction using sequence specific primers complements current DNA-based Rh system typing strategies and is superior in the detection of variant RHD alleles. PMID- 9354821 TI - The KEL24 and KEL14 alleles of the Kell blood group system. AB - BACKGROUND: The Kell blood group system consists of at least 21 antigens, which may be classified into five sets of alleles and at least 10 independently expressed antigens. The molecular basis of four of the five sets of alleles has been described; point mutations in KEL leading to amino acid substitutions characterize the alleles. In this study, the point mutation associated with the remaining allele, KEL14/KEL24, was determined. STUDY DESIGN AND METHODS: The 19 exons of KEL were amplified from genomic DNA by a polymerase chain reaction (PCR) procedure. The PCR products were sequenced. DNA sequences from unrelated KEL:14,24 and KEL:-14,24 individuals were compared to the DNA sequence of the common KEL:14,-24 phenotype. RESULTS: DNA from the KEL:14,24 person yielded both G and C at nt 659, indicating an Arg and Pro polymorphism in amino acid residue 180 of Kell protein. DNA from the KEL:-14,24 person had a G659C mutation in exon 6, indicating an Arg180Pro substitution. The G659C change introduces an Hae III restriction enzyme site, which was used to confirm the base mutations by restriction fragment length polymorphism analysis of the PCR products. CONCLUSION: A G659C mutation, predicting an Arg180Pro change in Kell protein, is associated with the KEL14/KEL24 allele. PMID- 9354822 TI - Prestorage inline filtration of whole blood for obtaining white cell-reduced blood components. AB - BACKGROUND: Preliminary studies have indicated that the inline filtration of whole blood is a feasible method of obtaining white cell (WBC)-reduced packaged red cells (RBCs) and WBC-reduced fresh-frozen plasma (FFP) while using only one filter. STUDY DESIGN AND METHODS: An inline WBC-reduction filter, specially designed for this purpose and integrated in a "top-top" system, was used in the preparation of 24 units of WBC-reduced RBCs (RBC-F) and FFP (FFP-F) in each of two transfusion centers (Vienna and Gottingen). Twelve conventionally prepared units of RBCs (RBC-C) and FFP (FFP-C) served as controls. WBC contamination was assessed in each unit with the Nageotte chamber. Several coagulation measures were evaluated by using standardized test systems. RESULTS: The median WBC contamination in RBC-F was 27,000 per unit in Vienna and 50,000 in Gottingen. In FFP-F, the median WBC contamination was 13,000 (Vienna) and 31,000 (Gottingen) per unit. Coagulation factors I, V, VIII, and XI in FFP-F were not different from those in FFP-C. In addition, markers for the activation of coagulation and fibrinolysis--that is, factor XIIa, prothrombin fragments, thrombin-antithrombin complexes and fibrinogen degradation products--were not greater in FFP-F. CONCLUSION: Blood components prepared from inline-filtered whole blood meet the standards for WBC-reduced RBCs and FFP. The protein profile of FFP-F is not altered, and markers for the activation of coagulation and fibrinolysis show no increase. PMID- 9354823 TI - Comparison of COBE white cell-reduction and standard plateletpheresis protocols in the same donors. AB - BACKGROUND: It is necessary to protect patients from white cell (WBC)-caused side effects of platelet transfusion by reducing the WBC contamination in single-donor platelets. STUDY DESIGN AND METHODS: A new COBE Spectra WBC (leuko)-reduction system (LRS) was compared to the COBE standard plateletpheresis (standard) procedure. Each of 62 donors underwent plateletpheresis under the two protocols (LRS and standard). The collection efficiency and WBC contamination in the components collected using the techniques were compared. Platelets were counted in a cell counter and WBCs were counted using two full grids of a Nageotte chamber. RESULTS: The preseparation and postseparation numbers for red cells, WBCs, and platelets as well as the number of collected platelets were not different in the two techniques. Collection efficiency in the LRS procedures was 96.2 +/- 13.0 percent of that in the standard procedures. Median WBC contamination in the platelet components was 10,160 per LRS procedure and 56,500 per standard procedure. The purity of the LRS components was significantly improved (p = 0.001), as seen in a comparison of the WBC numbers in components per procedure after log10 transformation (LRS: 0.096 +/- 0.195 x 10(6); standard: 0.390 +/- 1.075 x 10(6)). CONCLUSION: These data suggest that the LRS procedure produces platelet concentrates with a collection efficiency that is comparable to that obtained with the standard technique and with a residual WBC content that satisfies even the most stringent criteria for filtered platelets. As this purity can be achieved without platelet loss or alteration, conventional fiber filtration no longer seems necessary or useful in this type of single-donor platelet component. PMID- 9354824 TI - Comparison of CD34+ cell numbers and colony growth before and after cryopreservation of peripheral blood progenitor and stem cell harvests: influence of prior chemotherapy. AB - BACKGROUND: Quantitative determination of hematopoietic progenitor cells is a major issue in peripheral blood progenitor and stem cell collection and transfusion, although the extent is still an object of discussion. STUDY DESIGN AND METHODS: In 116 leukapheresis collections from 42 patients, immunophenotyping for CD34+ cells, evaluation of in vitro proliferative capacity by a colony forming unit-granulocyte-macrophage (CFU-GM) assay, and viability assessment by trypan blue exclusion were performed before and after storage in liquid nitrogen at -196 degrees C. RESULTS: Before storage, the median number of CD34+ cells was 1.46 x 10(6) (range, 0.01-54.05 x 10(6)) per kg of body weight (BW). There was no significant difference between precryopreservation and postcryopreservation numbers. The median number of CFU-GM was 2.25 x 10(5) (range, 0.02-157.49 x 10(5)) per kg of BW before cryopreservation and significantly (p < 0.001) lower, 0.83 x 10(5) (range, 0-220.36 x 10(5)) per kg of BW, after cryopreservation. The correlation coefficient of prestorage and poststorage values was 0.92. The median ratio of poststorage and prestorage values was 42.3 percent (0-304.8%). Male patients who underwent intense chemotherapy (> 5 cycles) showed a significantly lower ratio of postcryopreservation and precryopreservation CFU-GM values than other patients (p = 0.0047). A strong linear correlation was determined between the number of CD34+ cells per kg of BW and the number of CFU-GM per kg of BW before and after cryopreservation. A viability below 50 percent predicted a high loss of in vitro proliferative capacity, while a viability above 50 percent did not correlate with a high ratio of CFU-GM from after and before cryopreservation. CONCLUSION: A good correlation between the variables used for characterization of peripheral blood progenitor cells--the number of CD34+ cells and the number of CFU-GM--was observed. Viability assessment by trypan blue exclusion does not seem to be a substitute for assays evaluating in vitro proliferative capacity. PMID- 9354825 TI - Preoperative red cell production in patients undergoing weekly autologous blood donation. AB - BACKGROUND: Modest autologous blood donation programs involving weekly phlebotomy and threshold hematocrits for blood donation higher than 33 percent are frequently used in patients scheduled for elective cardiac surgery. This study was performed to determine the gain in red cells (RBCs) obtained with such a program. STUDY DESIGN AND METHODS: The blood bank and medical records of 225 adult patients (194 men, 31 women; mean age, 57 years [range, 18-77]) who donated blood for autologous use in elective cardiac surgery during a 3-year period were reviewed. Preoperative RBC production was estimated by the total volume of RBCs donated minus the change in circulating RBC volume between the first donation and the day before surgery. RESULTS: A total of 604 blood units were donated (2.7 units/patient; range, 1-3). The mean volume of RBCs donated was 522 mL (range, 171-732). Mean RBC production (over baseline RBC production) was 351 mL (range, 9 719), or 19 percent (range, 0.5-40) of the circulating RBC volume at baseline. CONCLUSION: A modest autologous blood donation program using three phlebotomies at weekly intervals and a threshold hematocrit for blood donation of 36 percent yields an average of 351 mL (range, 9-719) of RBCs. This is equivalent to 2 units (range, 0.5-4) of allogeneic packed RBCs at 180 mL per unit. PMID- 9354826 TI - Unexpected citrate toxicity and severe hypocalcemia during apheresis. AB - BACKGROUND: The citrate anticoagulant used during apheresis procedures is considered a safe medication because it is rapidly metabolized by the donor. However, acute, life-threatening hypocalcemia is possible if the infusion rate of citrate is increased. CASE REPORT: A 54-year-old woman with metastatic breast cancer, but otherwise in good health, had just begun a fifth collection of hematopoietic peripheral blood progenitor cells by leukapheresis. The instrument's self-loading apheresis kit was primed uneventfully. Seven minutes into the procedure, the patient developed signs and symptoms suggesting severe hypocalcemia, including muscle spasms, chest pain, and hypotension. The citrate bag was discovered to have emptied, and a section of the anticoagulant tubing was protruding outside of the rotary pump. The patient's ionized calcium level was 0.64 mmol per L (normal range, 1.18-1.38 mmol/L). In subsequent experiments where the anticoagulant tubing was either improperly loaded at the outset or partially pulled out of the rotary pump, no instrument alarms sounded. CONCLUSION: Citrate toxicity and life-threatening hypocalcemia can occur if the anticoagulant line of an apheresis instrument is not properly housed in its rotary pump or becomes disengaged during the procedure. Instrument manufacturers are encouraged to consider designs that allow the direct measurement of the volume of citrate delivered. In the interim, periodic visual and tactile confirmation of tubing placement during apheresis procedures is prudent. PMID- 9354827 TI - The effect of transfusion on cardiac function in patients with chronic anemia. AB - BACKGROUND: The deteriorating cardiac function of patients with chronic anemia may be improved with transfusion. The effect of transfusion on cardiac function was evaluated in patients with chronic anemia. STUDY DESIGN AND METHODS: In a prospective study, ejection fraction (EF) was determined before and after transfusion in 41 patients with chronic anemia. The results were compared and analyzed. RESULTS: The volume of red cells transfused and the levels of pretransfusion hemoglobin, hematocrit, and red cell, white cell, and platelet counts did not affect the posttransfusion EF, whereas the pretransfusion EF of the right or left ventricle inversely affected the posttransfusion change in EF in the respective ventricle (p < 0.001 and r = -0.5022; p = 0.01 and -0.3917, respectively). There was no significant difference in the change in EF in the right and left ventricles. CONCLUSION: Transfusion produced little immediate effect on cardiac function, but did change the EF to an extent that aided cardiac function in chronic anemia patients. The pretransfusion EF itself, but not the degree of anemia or volume of red cells transfused, affected the posttransfusion change in EF. PMID- 9354828 TI - A standardized method for calculating blood loss. AB - BACKGROUND: The estimation of blood loss for a surgical procedure is both poorly reproducible and typically underestimated. Therefore, comparison of surgical transfusion outcomes such as blood loss and amount of blood transfused from one institution to another, or from one surgeon to another, is difficult. Recently, mathematical modeling has contributed to our understanding of transfusion strategies. STUDY DESIGN AND METHODS: A mathematical model of blood loss for a surgical hospitalization was developed on the basis of recently described mathematical principles for blood loss and hemodilution. The model was designed so that the calculation of blood loss would be based on easily measured factors such as the patient's blood volume, the number and type of red cell units transfused, the initial hematocrit, the discharge hematocrit, the transfusion trigger, the volume of intraoperatively salvaged blood transfused, and the amount of hemodilution performed. The calculated blood loss was then compared with the intraoperative blood loss actually estimated by the anesthesiologist in 250 consecutive patients who underwent radical retropubic prostatectomy. RESULTS: The mathematical equations were placed in a computer model to allow rapid calculation of a particular patient's blood loss. Figures were derived from the computer modelling to facilitate rapid manual calculation of the blood loss. There was a significant relation (p < 0.001) between the calculated blood loss for the hospitalization and the estimated intraoperative blood loss. However, the calculated blood loss was on average 2.1 times the intraoperative blood loss estimated by the anesthesiologist. CONCLUSION: The use of such mathematical modeling to rapidly estimate a patient's blood loss has the potential to allow ready, objective comparisons between sites and even surgeons. It also allows for a more judicial and informed decision as to what (if any) blood should be available or what blood-conservation techniques should be employed for a specific patient. PMID- 9354829 TI - A survey of blood component use in a German university hospital. AB - BACKGROUND: There are no recent studies on transfusion practice and blood use with regard to diagnoses of European recipients. We conducted a survey of blood component use, including packed red cells, fresh-frozen plasma, and platelets, in an acute-care university hospital in the Greater Nurnberg area. STUDY DESIGN AND METHODS: A survey was carried out of blood component transfusion at a university hospital (Erlangen, Germany) between June 1994 and May 1996. Transfused units were listed by broad diagnostic categories formed from principal diagnoses of the recipients according to the International Classification of Diseases, Ninth Revision. RESULTS: Among 100,497 discharged patients, 6,590 patients who received transfusion (6.6%) are represented in this survey. Of 28,440 red cell units and 8,592 fresh-frozen plasma units, 72.4 percent and 66.9 percent, respectively, were used in patients with neoplastic diseases, circulatory system diseases, or disorders of the digestive system. Of 2704 platelet units, 78.1 percent were transfused to patients with neoplastic or gastrointestinal diseases or diseases of blood-forming organs. These four diagnostic categories accounted for 77.7 percent of all costs of transfusion therapy. Males received 60.1 percent of all blood components transfused, and patients less than 65 years old received 68.0 percent. CONCLUSION: This survey provides information on blood component usage in a German university hospital. It demonstrates the concentration of today's blood utilization among a few diagnostic categories. The study shows that detailed information on local blood use may be obtained quickly by using data available from transfusion services and medical record departments. This information is relevant for quality management of transfusion practice, cost analyses and for planning local and regional blood donation programs. PMID- 9354830 TI - Principles of blood irradiation, dose validation, and quality control. PMID- 9354831 TI - The essential standards. PMID- 9354832 TI - TRANSFUSION: the first decade: Volume 10. PMID- 9354833 TI - The sickle cell hemolytic transfusion reaction syndrome. PMID- 9354834 TI - The sickle cell hemolytic transfusion reaction syndrome. PMID- 9354835 TI - Monoclonal anti-D is not effective in the treatment of chronic autoimmune thrombocytopenic purpura. PMID- 9354836 TI - Increased expression of i on HEMPAS red cells: a flow cytometric study. PMID- 9354837 TI - Macrocephaly with cutis marmorata, haemangioma and syndactyly--a distinctive overgrowth syndrome. AB - We describe nine children with a similar pattern of features including macrocephaly and cutis marmorata telangiectatica congenita. All were large at birth and had a distinctive capillary haemangioma involving the philtrum and upper lip. The seven who survived all developed hydrocephalus and had developmental delay. Six developed body asymmetry and three had internal arteriovenous malformations. Syndactyly of the second and third toes and/or the third and fourth fingers or toes was commonly seen. All of the cases were sporadic. This condition is easily recognizable and should be considered in the differential diagnosis of patients presenting with overgrowth and macrocephaly. PMID- 9354838 TI - The one bone spine: a failure of notochord/sclerotome signalling? AB - Three cases of extensive vertebral fusions with absent clivo-axial angle are presented. The 'bone-within-bone' appearance in two patients with almost complete fusion of the spine suggested ossification of the notochord and perinotochordal sheath. On the basis of the radiological appearances and the results of recent molecular genetic studies on vertebrate embryos, the suggested time of segmentation failure along the axis of the craniovertebral junction and between vertebrae is the third to fifth week of gestation. The possible roles of the Pax 1 gene and of signalling between notochord and sclerotome are discussed, concluding that an early defect of the notochord may be responsible for this type of failure of segmentation. Indications for surgery in these cases included cord compression with brachialgia and 'chin-on-chest' deformity causing severely restricted visual fields. A critical review of clinical lessons learned in the operative treatment is presented. PMID- 9354840 TI - Extreme microcephaly, severe growth and mental retardation, flexion contractures, and ichthyotic skin in two brothers: a new syndrome or mild form of Neu-Laxova syndrome? AB - Two brothers with congenital microcephaly, growth and mental retardation, flexion contractures, dorsal edema of hands and feet, and ichthyotic skin changes are described. Mild manifestations of Neu-Laxova syndrome have to be considered but long survival and only mild intrauterine growth retardation not described in this syndrome may be evidence of a different condition. PMID- 9354839 TI - Escher-Hirt syndrome. AB - A mother and her two daughters are reported with bilateral conductive deafness due to incudo-stapedial abnormalities, and microtia with thickened ear lobes. This pattern of abnormal findings, transmitted with an autosomal dominant mode of inheritance, is characteristic of the Escher-Hirt syndrome. One of the daughters died from an additional cardiac malformation (VSD). Anomalies of the middle ear were demonstrated in the surviving patients by computed tomography. Differential diagnosis with other genetic syndromes associated with deafness, and possible therapeutic approaches are discussed. PMID- 9354841 TI - Osteogenesis imperfecta with arthrogryposis multiplex congenita (Bruck syndrome)- evidence for possible autosomal recessive inheritance. AB - We report a son and a daughter of a first cousin Pakistani marriage who both have osteogenesis imperfecta and the son in addition has arthrogryposis multiplex congenita. Bruck [(1897): Dtsch Med Wochenschr 23: 152-155] first reported the case of a boy who had multiple fractures and joint ankylosis, subsequently only one sibship with three affected cases and seven sporadic cases have been reported to our knowledge. On the basis of consanguinity this suggests that the association of osteogenesis imperfecta and arthrogryposis multiplex congenita is inherited in this family as an autosomal recessive condition with variable expression. PMID- 9354842 TI - A female patient with neurological, facial, digital and renal abnormalities: another case of the neurofaciodigitorenal (NFDR) syndrome? AB - We report a female patient with severe mental retardation and multiple congenital anomalies. These consist of unusual facies (grooved, nasal tip, ptosis, malformed auricles), abnormal digits, and congenital heart and renal defects. These findings strongly resemble the NFDR syndrome, first described by Freire-Maia et al. PMID- 9354843 TI - Are t(X;Y) (p22;q11) translocations in females frequently associated with Madelung deformity? AB - We report a female with a de novo 46,X,der(X)t(X;Y) (p22;q12) translocation who presented with short stature, mild clinical features of Turner syndrome and a Madelung deformity. It appears that some particular radiological and/or clinical skeletal features are common in females carrying X-Y translocation. Based on the corresponding papers and on clinical findings of our patient we discuss the significance of Madelung deformity encountered in X-Y translocations, dyschondrosteosis and Turner syndrome. PMID- 9354844 TI - Spondylocostal dysostosis associated with a 46, XX,+15,dic(6;15)(q25;q11.2) translocation. AB - We describe a female neonate with spondylocostal dysostosis and a translocation resulting in monosomy for the region 6q25-->qter and trisomy for the region 15q11.1-->pter. The finding of a Mendelian disorder with a chromosomal abnormality may help in the localization of the gene(s) involved in this disease. PMID- 9354845 TI - Anencephaly with holoprosencephalic facies due to ring chromosome 18. AB - An anencephalic infant with holoprosencephalic facies and ring chromosome 18 [r(18)] is reported with review of the pertinent literature. Although the association of anencephaly and holoprosencephalic facies is well established, this is the first instance of an accompanying karyotypic abnormality. Review of other r(18) cases suggests that this is not a coincidental finding. Karyotype analysis appears warranted in cases of anencephaly with holoprosencephalic facies. PMID- 9354846 TI - Fetal akinesia deformation sequence in a highly developed acardius twin. AB - We report a case of a holoacardius twin with extremely advanced development of the head, face, upper and lower limbs in the absence of all thoracic and upper abdominal viscera and associated with intestinal and anal atresia. The malformed fetus also had craniofacial abnormalities, hydrops, cystic hygroma of the neck, arthrogryposis and pterygia. The monozygous co-twin was found to be normal. The association of acardia with the typical characteristics of the fetal akinesia deformation sequence has not been previously described in the literature. PMID- 9354847 TI - Transverse limb defects, holoprosencephaly and neuronal heterotopia--a new syndrome? AB - We report an 18 week old fetus with four limb terminal transverse defects, holoprosencephaly with neuronal heterotopia and facial dysmorphism. We believe that this combination of features has not previously been described. PMID- 9354848 TI - Severe cardiac defect in a patient with the OEIS complex. AB - The OEIS complex is an association of fetal malformations including omphalocele, exstrophy of the cloaca, imperforate anus and spinal defects. We present a fetus with the OEIS complex in combination with a cardiac defect. Until now very few cases with this combination have been described. PMID- 9354849 TI - Craniofacial anomalies, severe cerebellar hypoplasia, psychomotor and growth delay in a child with congenital hypothyroidism. AB - We describe a 4-year-old boy affected by congenital hypothyroidism (CH) with the unusual association of brachycephaly, large and poorly structured ears, bilateral convergent strabismus, pectus carinatum and slight scoliosis, psychomotor delay, growth retardation and a severe hypoplasia of the right cerebellar hemisphere and vermis. Given the finding of unilateral cerebellar pathology this unusual association might be explained by an insult in utero--more environmental than genetic. However, to the best of our knowledge, a relationship between CH and cerebellar anomalies has not been previously reported. PMID- 9354850 TI - Progressive hydrocephalus in Noonan syndrome. PMID- 9354851 TI - Diabetic nephropathy and genetic alterations of the renin-angiotensin-system. PMID- 9354852 TI - Angiotensin I-converting enzyme-gene-polymorphism: relationship to albumin excretion and blood pressure in pediatric patients with type-I-diabetes mellitus. AB - The purpose of this study was the evaluation of a predictive genetic marker for nephropathy and hypertension in patients with type-I-diabetes mellitus (IDDM). The study was performed on 247 pediatric patients with IDDM. The mean age was 15.5 years (range 3.1-29.3), the mean duration of diabetes was 7.6 years (range 0.1-25.7). Age-related blood pressure and nocturnal albumin excretion rate were compared with the insertion/deletion-(I/D) polymorphism of the angiotensin-I converting enzyme gene. The genotype distribution did not differ significantly between IDDM patients (ID 48%, D 28%, I 24%) and the control group (ID 44%, D 37%, I 19%). Neither in the entire group, nor in patients with IDDM for more than 5 years, was a correlation found bet-ween allele distribution and albumin excretion rate. No correlation was found between genotype and blood pressure. When patients with a chronological age above 12 years were analysed separately, the genotype distribution between the groups with normal and elevated blood pressure showed no significant difference. The previously reported association of the I/D-polymorphism with nephropathy could not be confirmed in this study. The development of microalbuminuria, nephropathy and hypertension will be followed in our pediatric patients. PMID- 9354853 TI - Changes in the signalling status of the small GTP-binding proteins Rac and Rho do not influence insulin-stimulated hexose transport. AB - Post-receptor signalling molecules that convey the signal from the activated insulin receptor to the actual process of Glut4 translocation and hexose uptake are poorly understood. Various studies have suggested a requirement of the lipid kinase phosphatidylinositol-3 kinase (PI3-kinase) in this process. PI3kinase regulates the activation status of the small GTP-binding protein Rac which, in turn, is able to activate another G-protein Rho. Rac and Rho are known to regulate the structure of the membrane- and cytoplasmic actin-cytoskeleton. We have examined whether Rac and Rho transfer the signals generated by PI3kinase towards insulin-stimulated hexose uptake. For that purpose, we expressed in 3T3 L1 adipocytes the dominant-negative mutant of RacN17 using vaccinia virus mediated gene transfer. The expression levels of the RacN17 protein were monitored by Western blotting. The abrogation of endogenous Rac signalling by expression of RacN17 was inferred from the observed loss of arachidonic acid release in response to insulin. Basal and insulin-stimulated hexose transport were not affected by expression of the RacN17 mutant. A possible contribution of Rho.GTP to stimulation of hexose uptake was examined by pre-incubation of adipocytes with lysophosphatidic acid (LPA). We observed a profound effect of LPA on the structure of the cytoskeleton and on the phosphorylation of Focal Adhesion Kinase (p125FAK), indicating that 3T3-L1 adipocytes respond to LPA and that Rho was activated by LPA. However, no effect was detected on the basal or on the insulin-stimulated hexose transport. We conclude that Rac and Rho are unlikely to be involved in insulin-stimulated hexose transport, suggesting a possible contribution of other signalling pathways, downstream of PI3kinase to this process. PMID- 9354854 TI - Evidence for binding of in vitro glycated low-density lipoproteins by fructosyllysine-specific sites on macrophages and U937 monocyte-like cells. AB - Early development of arteriosclerosis is a main late complication of diabetes mellitus. Although its uptake by LDL receptors is impaired, glycated LDL is thought to play a role in foam cell formation from macrophages. In the present study we show binding of glycated LDL (8-9 mol fructosyllsine/mol apo B) to macrophages and to the monocyte-like cell line U937. The binding involves fructosyllysine-specific binding sites, as well as LDL and scavenger receptors. Fructosyllysine and glycated albumin were competitors for binding of 125I labelled glycated LDL by macrophages and U937 cells. Scatchard analysis of binding data using a two ligands binding model showed a linear plot with Ka = 2.6 x 10(7) M-1 for U937 cells, which lack scavenger receptors. On U937 cells only the 200 kDa fructosyllysine-specific receptor protein and the 165 kDa LDL receptor were involved in binding glycated LDL as evidenced by ligand blotting. U937 cell uptake and degradation of glycated LDL was mediated by fructosyllysine specific and LDL receptors. Binding of glycated LDL by macrophages via fructosyllysine-specific sites could be demonstrated in only 6 from 35 rats investigated, indicating that the receptor is not expressed in each individual. Whether the fructosyllysine-specific receptor mediated pathway is relevant for uptake and degradation of glycated LDL in vivo is yet to be determined. PMID- 9354855 TI - Relationship between follicular fluid steroids and tissue renin concentrations and secretion rates in bovine ovaries. AB - The relationship between follicular fluid concentrations of oestradiol (E2) and progesterone (P4), and follicular wall tissue renin concentrations and in-vitro secretion rates was investigated. The follicular fluid concentrations of prorenin correlated negatively with oestradiol and the E2/P4 ratio, and positively with progesterone in accordance with earlier findings. The high concentrations of active renin and prorenin in the follicular wall tissue, and the secretion rates of both active renin and prorenin all correlated positively with oestradiol and the E2/P4 ratio, but negatively with progesterone. These findings suggest that atretic follicles have higher follicular fluid renin concentrations but lower tissue renin concentrations and in-vitro secretion rates than healthy non-atretic follicles. The high follicular fluid prorenin concentrations in atretic follicles are most likely caused by a high prorenin secretion rate earlier in the oestrus cycle. The existence of relationships between the follicular fluid steroids and the follicular wall concentrations and secretion rates of active renin and prorenin indicates an interaction between the reninangiotensin system and steroidogenesis, but it may also reflect regulation by common factors, such as gonadotropins. PMID- 9354856 TI - Different mechanisms mediate the in vivo aldosterone and corticosterone responses to 5-bromo-2'-deoxyuridine in rats. AB - The subcutaneous injection of 5'-bromo-2' deoxyuridine (BrdU) was found to raise the plasma concentrations of ACTH, aldosterone and corticosterone in rats. The aldosterone response was observed at a lower dose of BrdU and lasted for a longer period than those of ACTH and corticosterone (1.25 versus 2.50 mg/100 g body weight; 48 versus 24 h). Corticosterone response to BrdU was partially reversed by the ACTH-receptor antagonist corticotropin-inhibiting peptide (CIP), and aldosterone response by the arginine vasopressin (AVP) V1-receptor antagonist [amino-Pen1, Val4,D-Arg8]-vasopressin (AVP-A). The angiotensin-II (ANG-II) receptor antagonist [Sar1, Val5, Ala8]-ANG-II (SAR) was ineffective. CIP, AVP-A and SAR, when administered alone, did not alter basal levels of ACTH, aldosterone and corticosterone. In light of these findings the following conclusions can be drawn: (i) BrdU stimulates the hypothalamo-pituitary-adrenal axis in rats, and this effect may influence the results of cell-kinetics studies carried out with the BrdU-labelling technique, especially in those tissue that are highly responsive to glucocorticoids (e.g. pituitary, adrenal and lymphatic tissues); and (ii) different mechanisms underlie the aldosterone and corticosterone secretagogue effects of BrdU, the former being at least in part dependent on the stimulation of AVP release and the latter on the rise in ACTH secretion. PMID- 9354857 TI - Expression of a functional tagged human thyrotropin receptor in HeLa cells using recombinant vaccinia virus. AB - We report a new effective system for expression of FLAG and six histidine tagged TSH receptor in mammalian cells using recombinant vaccinia virus. HeLa cells infected with recombinant virus produced large amounts of human TSH receptor of approximately 150,000 functional molecules per cell. This value is one to two orders of magnitude higher than those in thyroid cells and is comparable with receptor number of the best stably transfected mammalian cell clones previously described. Vaccinia virus produced TSH receptor was able to bind TSH (Kd of 2.1 +/- 0.1 x 10(-10) M) and Graves'disease autoantibodies. TSH caused an increase of the intracellular cAMP level in infected HeLa cells in a concentration-dependent manner, demonstrating the coupling of expressed recombinant TSH receptor to the cAMP second messenger system of the cells. 6His-tagged recombinant TSH receptor was immobilized on Ni2+ nitrilotriacetic acid-agarose. Bound receptor was fully functional, interacting with both TSH and Graves' disease autoantibodies in patient sera. The solid phase bound TSH receptor technique provides a new and simple method for the diagnosis of autoimmune diseases. PMID- 9354858 TI - Reproductive hormonal responses to maximal exercise in endurance-trained men with low resting testosterone levels. AB - A cross-sectional study was conducted to compare the changes from rest and in response to a maximal exercise bout for select reproductive hormones between age matched groups of endurance trained (ET; distance runners) men with low resting testosterone and untrained (UT) men. Both ET and UT men completed two evaluation sessions: (a) resting hormonal profiling, and (b) a maximal treadmill exercise test to exhaustion. Serial blood samples were taken for four hours at each of the evaluation sessions. Resting and exercise hormonal concentrations were plotted and the area under the response curve (AUC) measured. Percentage change in AUC values were also calculated and compared (exercise vs. resting AUC values). Resting testosterone (16.6 +/- 2.4 vs. 23.9 +/- 3.1 nmol x 1(-1)) and prolactin (3.3 +/- 1.4 vs. 6.0 +/- 2.0 micrograms x 1(-1)) concentrations in the ET men were significantly lower (p < 0.05) than those in the UT men. All other resting hormonal levels did not differ between the groups (p > 0.05). Exercise produced a significant increase (p < 0.05) in the ET men for testosterone, LH, and prolactin AUC values, when compared to resting values. In the UT men the only significant change was a reduction (p < 0.05) in the exercise LH AUC versus the resting AUC value. AUC percentage change values showed between-group differences (p < 0.05) for testosterone, LH and prolactin. The level of change in each of these hormones was found to be greater in the ET than UT group (approximately 20 to 75%). The hormonal changes of the UT men were viewed as "control--reference" responses within a functioning hypothalamo-pituitary-testicular regulatory axis; therefore, it was concluded that the ET men displayed an "atypical" response to exercise to that of UT men relative to this axis. PMID- 9354859 TI - Determination of free and conjugated oestrogens in peripheral blood plasma, feces and urine of cattle throughout pregnancy. AB - In order to further characterize oestrogen production and metabolism during bovine pregnancy, free (f) and conjugated (c) estrone (E1), total free and conjugated oestrogens (tfcOe) and total free oestrogens (tfOe) were determined as marker oestrogens in blood plasma respectively in urine and feces of 10 pregnant cows. For the determination of individual oestrogens blood, urine and feces samples of days 240, 200, 160, 100, 60, 30, 10 and 5 prior to parturition were pooled and the free, sulfo (sc)- and glucuconjugated (gc) forms of E1, 17 beta estradiol (E2 beta) and 17 alpha-estradiol (E2 alpha) were obtained following differential enzyme hydrolysis and separation by HPLC; hormone assay was by established RIA-procedures. FE1 and cE1 concentration in blood plasma, tfOe in feces and tfcOe in urine showed a similar pattern. A first rise occurred between days 110 and 120 of pregnancy, an additional overproportional rise commenced at around days 230-250. Highest concentrations were measured in feces (tfOe ca. 500 ng/g 1 day a. p.), followed by urine (tfcOe ca. 3.5 ng/mosmol 2 days a. p.) and blood plasma (fE1 ca. 8 nmol/l and cE1 ca. 20 nmol/l 2 days a. p.). Determination of individual oestrogens in blood plasma revealed that fE2 beta and fE2 alpha could only be found 10 days a. p. while the conjugated forms could already be detected on days 100 and 160 a. p. With 62% E1 was the dominant oestrogen, followed by E2 alpha (37%) and E2 beta (1.0%); E1 occurred predominantly as sulfate, E2 alpha and E2 beta predominantly as glucuronide. Main metabloite in feces was fE2 alpha (56.7%), followed by fE2 beta (32%) and fE1 (11.3%); conjugated oestrogens were not detected. Main metabolite in urine was scE1 followed by gcE2 alpha and gcE2 beta. ScE2 alpha and scE2 beta were not detected or were present in small quantities only. Hormonal changes over time were highly significant. Main product of placental oestrogen synthesis is scE1, the concentrations of f and c E2 beta and E2 alpha in plasma largely result from oestrogen metabolism and enterohepatic circulation. PMID- 9354860 TI - Involvement of tryptophan in the structural alterations of the rat ovarian LH/hCG receptor. AB - Treatment of the rat ovarian membrane-bound and Triton X-100 solubilized LH/hCG receptor with the tryptophan-specific reagents N-bromosuccinimide (NBS) and 2 hydroxy-5-nitrobenzyl bromide (HNB-Br) resulted in inactivation of the receptor to bind hCG. Fluorescence quenching studies indicated that oxidation of tryptophan residues by NBS decreased the accessibility of fluorophores for acrylamide. Preceding binding of hCG to receptor sites was found to protect fluorophores from NBS action. Modification of tryptophan residues was associated with alteration in the rigidity of ovarian membranes and with destabilization of the LH/hCG receptor structure. The results suggest that tryptophan residue is essential for hCG binding to the receptor. PMID- 9354861 TI - Counter current transfer of beta-endorphin in the perihypophyseal cavernous sinus -carotid rete vascular complex of sheep. AB - This study was performed to answer the question of whether counter current retrograde transfer of beta-endorphin in the perihypophyseal cavernous sinus carotid rete vascular complex depends on the reproductive activity of sheep and if this transfer depends on membrane Na+ K+ ATP-ase blocking by ouabain. Sheep were anaesthetised and the jugular vein and the carotid artery were cannulated on both sides. Multielectrolitical liquids (Solfin, Polfa "Kutno", Poland): 500 ml of Solfin with heparin (25,000 IU), or Solfin with heparin and ouabain (Sigma, St. Louis, USA) in concentrations of 10(-5) or 10(-4) mol 1(-1) were infused into the brain through the carotid artery. Heparinized blood was collected through the carotid artery. After exsanguination, the head with the neck was removed. The isolated head was supplied with oxygenated, heated, autologous blood diluted with Solfin (4:1) without or with ouabain in concentration of 10(-5) or 10(-4) mol 1( 1). Blood pressure and temperature were measured throughout the duration of the experiment. During the experiment 125I-beta-endorphin (7.9 x 107 dpm) dissolved in 10 ml of Solfin was infused for 5 min (5 ml) into each cavernous sinus through the angularis oculi veins. Blood samples for radioactivity measurements were collected each min from the carotid rete (through the opposite carotid artery to the artery supplying the brain with arterial blood) and from both jugular veins. In all the experiments significant 125I-beta-endorphin radioactivity was found in arterial blood supplying the brain and hypophysis in the early luteal phase in sheep. No radioactivity was found (with the exception of one animal) in sheep during seasonal anoestrus. A blockage of Na+ K+ ATP-ase by ouabain administered during exsanguination and during head perfusion with dose of 10(-4) mol 1(-1) reduced beta-endorphin counter current transfer to arterial blood, but this effect was not evident with the dose of 10(-5) mol 1(-1). Increased blood pressure was observed in all the experiments with either dose of ouabain. PMID- 9354862 TI - Required research a disservice? PMID- 9354863 TI - Use of computers by family practice residency graduates. PMID- 9354864 TI - Alternative models of residency sponsorship. PMID- 9354865 TI - Orienting medical students in community-based teaching sites. PMID- 9354866 TI - Rewarding teaching in medical education. PMID- 9354867 TI - Skills that Iowa family physicians desire in a new physician partner. AB - BACKGROUND AND OBJECTIVES: The importance of specific skills in primary care continues to be debated. As a result, there is not consensus on which skills need to be stressed during residency training. Our project asked community-based family physicians to rate the importance of specific skills in a new family physician partner. METHODS: Data were collected through a cross-sectional survey of all active members of the Iowa Academy of Family Physicians. Participants were surveyed by mail, using a list of 83 skills pertinent to primary care. Physicians were asked to rate the importance of a new member of their practice having the individual skills on this list. RESULTS: A total of 546 family physicians (67%) completed questionnaires. Fourteen skills (seven cognitive and seven psychomotor) were reported to be "essential" or "very important" by at least 80% of the physicians. A total of 43 skills were rated as "essential" or "very important" by at least 50% of responding family physicians. Many of the hospital-based procedural skills, particularly those used in an intensive care setting, were rated as less important. The importance ratings of many skills were associated with the physicians' ages, size of their primary hospitals, and availability of other medical specialties. CONCLUSIONS: Family physicians tended to rate office based procedural skills, counseling skills, and management skills as "essential or very important" to their practices. These rating might be used to guide residency training in family practice. PMID- 9354868 TI - Attitudes of family practice residency program directors toward mandatory preemployment drug testing. AB - BACKGROUND AND OBJECTIVES: As health care institutions adopt policies on substance use and abuse and mandatory substance abuse testing in the workplace, applicants for house staff positions may become the subjects of testing as a requirement for acceptance into a residency program. This study attempted to learn what directors of family practice residency programs feel about mandatory preemployment drug testing and its effect on house staff recruitment. METHODS: We surveyed the directors of 420 US family practice residency programs, as listed by the American Academy of Family Physicians, in November 1994. All programs (community based, university affiliated, university based, and military) were included in the survey. RESULTS: A total of 308 (73%) program directors responded. Of these, almost half disagreed with mandatory substance abuse testing and felt it should not be a condition of acceptance for a house staff position. Eighty-eight percent believed that the existence of a policy did not hinder recruitment. None felt it was an enhancement. CONCLUSIONS: Preemployment drug testing for potential house staff remains a controversial issue, and it is unlikely that it will be universally implemented in the near future. PMID- 9354869 TI - Documenting resident procedure and diagnostic experience: simplifying the process. AB - BACKGROUND AND OBJECTIVES: The Residency Review Committee (RRC) requires documentation of family practice residents' procedural and diagnostic experiences. Further, hospital privileging is frequently based on documentation of prior clinical experience. Residency programs need a user-friendly (ie, resident-friendly) mechanism for collecting data and generating reports to document these experiences. This paper outlines a simplified, user-friendly method of documenting resident procedural and diagnostic experiences. METHODS: We developed a pocket-sized, optically scannable card for data input. This is coupled with a computerized database with report generation capability. The system is based on diagnostic clusters to further simplify the data input process. RESULTS: The system's setup costs are about $10,000. Annual maintenance and operational fees are about $5,000. After instituting the system, the number of residents submitting documentation information increased substantially. CONCLUSIONS: This system meets both RRC and potential clinical privileging requirements and provides a useful tool for guiding resident evaluation and developing appropriate training opportunities during the latter half of the residency. Simplified, accurate documentation may allow for comparisons among residents at various levels--program, state, and national. PMID- 9354870 TI - Home visit program for teaching elder abuse evaluations. AB - BACKGROUND AND OBJECTIVES: A home visit program was designed to teach family practice residents how to evaluate patients for elder abuse and capacity (the ability to make one's own decisions). METHODS: Residents assessed potential abuse victims reported to Arizona's Adult Protective Service (APS) in their homes. Written evaluations prepared immediately following each home visit were abstracted for diagnoses (including abuse), recommendations, and patient demographics. Follow-up surveys by APS case workers determined whether the home visit recommendations were accomplished. Graduates of the residency were surveyed about their perceptions of the educational value of the program and their practice characteristics. RESULTS: The residents evaluated 201 patients. The mean age was 77, and 73% of patients were female. Seventy-five percent were incapacitated, 65% of these because of dementia. Ninety-one percent were abused, and the types of abuse included neglect (69%), exploitation (20%), physical abuse (8%), and unknown (3%). Recommendations were accomplished in the majority of cases: medical advice (68%), services (65%), medical evaluations (58%), guardian (53%), and conservator (52%). Graduates who participated in this program (1985 1992) rated their ability to diagnose elder abuse and to assess the patient's home environment significantly higher than earlier graduates who did not participate in the program (1977-1984). Earlier graduates made more home visits and provided more statements for guardianship than later graduates. CONCLUSIONS: The home visit program gave residents exposure to a population of elderly who were abused, demented, and living at home. This program provided clinical substance to build an effective teaching experience and furnished APS with a needed service. PMID- 9354871 TI - Evaluating family practice residencies: a new method for qualitative assessment. AB - BACKGROUND AND OBJECTIVES: This study reports on a novel qualitative method for evaluating family practice training programs. Previous evaluation techniques have generally been quantitative in nature and have limited their scope to a few isolated elements of residency education. METHODS: A guest faculty, working in conjunction with local faculty, conducted a site analysis of an East Coast and a West Coast family practice residency. Multiple qualitative techniques were used, including participant observation, focus groups, long interviews, and analysis of key texts. Program strengths and weaknesses were analyzed, and a discrepancy model was used to compare program goals and ideals to the actual training realities. The analysis used a process of immersion/crystallization, and triangulation of the multiple data sources was achieved through repeated comparisons. RESULTS: This report focuses on the process of the evaluations, rather than on their content. In general, the sites have achieved most of their objectives, but notable limitations are present at both programs. This is particularly apparent in terms of multiple demands on faculty, the lack of a shared vision, and program isolation. CONCLUSIONS: Significant lessons were learned from these initial assessments, which can be used to further refine the method. Comprehensive qualitative reviews may provide unexpected insights and identify program limitations and strengths. PMID- 9354872 TI - The most common dermatologic problems identified by family physicians, 1990-1994. AB - BACKGROUND AND OBJECTIVES: Because all family physicians see numerous patients with dermatologic complaints, their education in skin disorders is important. Data are needed to help program directors know which areas of dermatology deserve the most time and emphasis. This study determined what types of skin problems family physicians most commonly diagnose. METHODS: Study researchers analyzed National Ambulatory Medical Care Survey data from 1990 to 1994 for dermatologic diagnoses. We then compared physicians specializing in family practice and its related fields (general practice, family practice sports medicine, and family practice geriatrics) with dermatologists and other physicians. RESULTS: The most common skin disorders diagnosed by family physicians were dermatitis (16.4% of all diagnoses), pyoderma (13.7%), wart (8%), tinea infection (5.4%), and epidermoid cyst (5.1%). The top 10 most common diagnoses accounted for 65% of all skin-related diagnoses, and the top 20 most common diagnoses accounted for 81.8%. Family physicians more commonly saw patients for infectious processes, infestations, and insect bites, while dermatologists were more likely to see patients for psoriasis, alopecia, and rosacea. CONCLUSIONS: Skin disorders diagnosed by family physicians differ considerably from those diagnosed by dermatologists. Because dermatologists do much of the dermatology teaching of family practice residents, it is important to recognize these dissimilarities to place emphasis on the proper areas of study. Some common or serious conditions, such as psoriasis and melanoma, are not often diagnosed by family physicians and also deserve attention in family practice training programs. PMID- 9354873 TI - Control, compliance, and satisfaction in the family practice encounter. AB - BACKGROUND AND OBJECTIVES: With the current emphasis on patient-centered interviewing, issues of control behavior have become an important facet for understanding effective physician-patient communication. In this study, we describe how verbal control behaviors are manifested during the clinical encounter and how these control patterns relate to patient satisfaction and compliance. METHODS: Videotaped encounters (n = 50) in a family practice residency clinic were transcribed and analyzed using the Relational Communication Control Coding Scheme. In addition, we surveyed patients to assess levels of compliance and satisfaction. RESULTS: Overall, patients showed assertive control patterns, and physicians manifested patterns of willingness to let patients take control of the conversation. The resulting outcomes showed that when physicians exhibited less control dominance, there was an increase in patient compliance and satisfaction. CONCLUSIONS: The control patterns discovered are consistent with patient-centered viewpoints that encourage the patient's expression of ideas, concerns, and expectations. Increased levels of patient satisfaction and compliance were found when patients more assertively participated in the clinical conversation. PMID- 9354875 TI - The health care delivery crisis in Haiti. AB - BACKGROUND AND OBJECTIVES: This article documents the history, politics, and economics that have contributed to a health care delivery crisis in Haiti and why family medicine will be crucial for the recovery of Haiti's health care. Since the United Nations intervention, there has been some improvement in health conditions. However, the embargo and political turmoil left little infrastructure on which to build. Developing family medicine, one of the priorities of the Ministry of Health, will reverse traditional forces that favor emigration and specialization and will provide the country with well-trained physicians who can treat most of the common health problems of Haiti. These common preventable and treatable problems are now contributing to short life expectancy and high infant mortality. While the ultimate responsibility for Haiti's health rests with Haitian health professionals, the country has an immediate need for international humanitarian assistance, particularly for general medical care. PMID- 9354874 TI - Patient-caregiver functional unit scale: a new scale to assess the patient caregiver dyad. AB - BACKGROUND AND OBJECTIVES: This study evaluated the reliability and validity of the Patient-Caregiver Functional Unit Scale (PCFUS), a new instrument to assess the stability or endurance of patient-caregiver dyads. METHODS: Patient-caregiver dyads were recruited from a nursing home (NH) (n = 38), a comprehensive geriatric assessment program (CGA) (n = 20), and an ambulatory medical clinic (controls) (n = 85). Caregivers were eligible if they assisted, or were available to assist, the patient with personal and instrumental activities of daily living, without pay. Data were collected by interviewer-administered questionnaires. Inter-rater and test-retest reliability were evaluated among the CGA sample. Validity was assessed by comparing PCFUS scores among the NH, CGA, and control groups and by correlation of PCFUS scores with other standardized caregiver burden measures. RESULTS: The PCFUS had excellent inter-rater and test-retest reliability. Mean PCFUS scores were significantly lower (ie, less stable patient-caregiver dyad) in NH than CGA and control caregivers. PCFUS scores were significantly associated with Burden Interview, Perceived Stress Scale, and Geriatric Depression Scale scores and risk factors for caregiver stress (eg, patient's cognitive impairment, disruptive behaviors). CONCLUSIONS: The PCFUS is a short, easily administered measure with good reliability and validity and is applicable to clinical and research settings. PMID- 9354876 TI - Second-trimester diagnosis of fetal cataract in a fetus with Walker-Warburg syndrome. AB - We report on the prenatal diagnosis of bilateral fetal cataract in the 17th week of pregnancy as the first ultrasonographic sign of a Walker-Warburg syndrome in a familial case. No other anomalies could be detected by ultrasonographic examination at that time. Both the bilateral fetal cataract and specific abnormalities of a Walker-Warburg syndrome were confirmed after the termination of the pregnancy in the second trimester. PMID- 9354877 TI - Antenatal diagnosis and treatment of fetal hypothyroidism. A report of two cases. AB - Fetal goiter in the presence of maternal Graves' disease can signify either hyperthyroidism or hypothyroidism in the fetus. Two patients with Graves' disease taking propylthiouracil were found to have a fetal goiter on a prenatal sonogram. Fetal blood sampling identified hypothyroidism in both instances. Administration of intraamniotic thyroxine resulted in rapid resolution of the goiter, normalization of subsequent fetal thyroid studies, and delivery of a euthyroid fetus. Cordocentesis allows accurate diagnosis and follow-up of fetal thyroid dysfunction and prompt initiation of appropriate therapy. PMID- 9354878 TI - Fetal tachycardia: intrauterine and postnatal course. AB - Eighteen consecutive cases of fetal tachycardia referred to the department of Pediatric Cardiology, Uppsala University, were studied retrospectively. All cases were detected at a routine visit at an antenatal clinic. None of the cases had a structural heart disease. Fetal supraventricular tachycardia was found in 8 cases and atrial flutter in 10 cases. In 7 cases, hydrops and heart failure were diagnosed. Antenatal treatment with digoxin, alone or in combination with other antiarrhythmic drugs, was needed in 15 cases. In 10 cases an obvious effect of the therapy was observed. No intrauterine deaths occurred. One infant died postnatally. At birth, 4 infants were in need of neonatal intensive care when delivered. Antiarrhythmic treatment was started in 13 cases postnatally. Late relapse of tachycardia was reported in 3 children. In 1 of these cases the prenatal tachycardia had resolved spontaneously and the infant was not treated antenatally nor during the neonatal period. Although fetal tachycardia is a serious condition, antenatal treatment in combination with careful monitoring and induction of delivery in cases with deteriorating fetal condition result in a satisfactory outcome for the majority of infants. However, there is a risk of late recurrence. PMID- 9354880 TI - Management of the fetus with a correctable malformation in Paris maternity units: evolution 1985-1994. AB - OBJECTIVE: To analyze the evolution of the management of delivery and neonatal care in a population of children with correctable malformations born in Parisian maternity hospitals during the period 1985-1994. METHODS: Data were collected by the Paris Registry of Congenital Anomalies from 400,000 births recorded in Parisian maternity hospitals over a 10-year period. Chromosomal anomalies were excluded. The evolution between the first period (1985-1989) and the second (1990 1994) was analyzed for the following indicators in the management of liveborn children: place of delivery; frequency of prenatal diagnosis; transfer to intensive care units, and mortality. RESULTS: More than 60% of the births of malformed children took place in public maternity hospitals where better management is offered. Most of them were prenatally diagnosed, except for esophageal and anorectal atresia for which the rate of prenatal diagnosis was low. For malformations with poor prognoses (diaphragmatic and abdominal wall anomalies), the rate of deliveries in public maternity hospitals reached about 90%, mostly in those with intensive care units. The evolution between the two periods was characterized by a quicker transfer to intensive care units, during the first day of life for most cases. Lethality during the first day, which was already low during the first period, decreased further. The early neonatal mortality rate decreased for cardiac anomalies, but not significantly. The prognosis remained poor for diaphragmatic anomalies: 49% of liveborn children died during the first week of life. PMID- 9354879 TI - Trisomy 21 placentas: histopathological and immunohistochemical findings using proliferating cell nuclear antigen. AB - OBJECTIVE: The cause of growth retardation in trisomy 21 and other autosomal trisomies is not known, but may be the result of defective cell proliferation, slowing of the cell cycle, or placental structural abnormalities. Abnormalities of the fetal cell cycle may be reflected in placental growth and can be detected using proliferating cell nuclear antigen (PCNA). METHODS: Twelve second-trimester and six third-trimester trisomy 21 placentas were examined histopathologically and stained immunohistochemically using antibodies to PCNA. Normal age-matched placentas were used as controls. RESULTS: The second-trimester trisomy 21 placentas all exhibited many large irregular hypovascular villi. The third trimester trisomy 21 placentas showed two patterns: (i) many large, irregular hypovascular villi, and (ii) relatively normal-appearing villi with only a few abnormal villi and focal hypervascularity. PCNA staining was significantly greater in second-trimester placentas when compared to third-trimester placentas for both trisomy 21 and controls. There was no significant difference in PCNA staining in trisomy 21 placentas when compared to the normal age-matched controls. CONCLUSIONS: PCNA staining indicates no significant differences in proliferation between normal and trisomy 21 placentas. Trisomy 21 placentas show villus abnormalities, including hypovascularity. PMID- 9354881 TI - Aspiration of giant hepatic cyst in the fetus in utero. AB - A large hepatic cyst was diagnosed in a fetus at 29 weeks of gestation by puncture and aspiration performed under ultrasonographic guidance. This procedure was repeated five times at about 2-week intervals, with a total of 1,446 ml of cystic fluid aspirated. A healthy female infant was delivered vaginally at 40 weeks of gestation. Her Apgar score was 9 at 1 min, and she weighed 2,790 g. On the 14th postnatal day, the hepatic cyst was punctured percutaneously under ultrasonographic guidance, and 119 ml of yellow fluid was aspirated. The cyst was subsequently not visible on computed tomography at 9 months. Surgery was not indicated in this infant, and the cyst was also not detected at the age of 21 month. Thus, treatment in utero appeared to have obviated the need for surgery. PMID- 9354882 TI - Fetal pulmonary artery Doppler waveform: a preliminary report. AB - OBJECTIVE: The study of fetal lung circulation by means of pulmonary Doppler investigation. METHODS: Pulmonary Doppler ultrasound obtained with color pulsed Doppler with a 3.5- to 5-MHz probe. Measure of resistance index and pulsatility index. PATIENTS: 47 pregnant women with singleton, between 18 and 39.5 weeks of gestation, were recruited to have pulmonary Doppler ultrasound. Seven fetuses had intrauterine fetal growth retardation (IUGR). Overall, 50 Doppler velocity waveforms were measured. RESULTS: Resistance and pulsatility index were measured in all patients and at each examination. Resistance index (0.86 +/- 0.03) and pulsatility index (2.46 +/- 0.34) were found to be stable during pregnancy. Pulmonary pulsatility index in IUGR fetuses (2.71 +/- 0.33) were found to be higher than those in normotrophic infants (p = 0.006), whereas no difference was found in resistance index between the same subgroups. Moreover, no difference was found in pulsatility index measurements between preterm small-for-gestational age fetuses and normotrophic fetuses measured between 36 and 39.5 weeks of gestation (2.68 +/- 0.31 vs. 2.49 +/- 0.28). CONCLUSION: Pulmonary resistance index is not statistically different between normotrophic and IUGR fetuses. In contrast, pulmonary pulsatility index is significantly higher in IUGR fetuses when compared to normotrophic fetuses. Pulmonary Doppler ultrasound should be evaluated in a larger trial and correlation between Doppler measurements and fetal lung maturation should be studied. PMID- 9354884 TI - First trimester fetal heart rate: response to chorionic villus sampling in the chromosomally normal fetus. AB - The objective of this prospective study was to evaluate the fetal heart rate before and after the procedure and whether this had any effect on the risk of pregnancy loss following chorionic villus sampling (CVS) and chromosomally normal pregnancies. Four hundred and seventeen pregnancies were evaluated. The heart rate before the procedure was similar for both male and female fetuses. Significant procedure-related changes in fetal heart rate occurred only with male fetuses undergoing the transabdominal CVS technique. Fetal heart rate decelerations following CVS were more common than accelerations. Fetal heart rate changes following CVS were not able to predict the pregnancies ending in spontaneous abortion. Transabdominal CVS had a lower loss rate after the procedure, compared to the transcervical technique (p = 0.0001). Small-for-date fetuses had a higher loss rate after the procedure compared to normal-sized fetuses (p = 0.001). PMID- 9354883 TI - In utero partial liver resection in the rabbit model: a study on fetal tissue regeneration. AB - In this study we developed a model of in vivo intrauterine partial liver resection in the fetal rabbit to analyze fetal liver regeneration. After intravenous anesthesia, 12 time-dated pregnant, California rabbits underwent a midline laparotomy and minimal hysterotomy at 24-25 days of gestational age. One fetus was exposed from each pregnant doe and the fetal liver was partially resected. Cesarean sections were performed 24, 48 and 72 h and 4 days after surgery. Three fetuses operated at 24 days of gestational age and 3 fetuses operated at 25 days were alive at retrieval. The fetuses and the sampled livers were weighed at retrieval and fetal liver weight showed a well-maintained value in all cases. Fetal livers were processed for the common histologic stains. Lymphocytes, polymorphonuclear leukocytes and phagocytes were counted from sections obtained in areas close to the edge of resection. Inflammatory cells showed a peculiar pattern of infiltration at different stages of repair, with a constantly increased number of phagocytes peaking 48 h after resection. Fetal liver seems to present a specific pattern of repair that differs from both the adult liver and other fetal tissues healing after injury. PMID- 9354885 TI - Large placental chorioangiomas as a cause of cardiac failure in two fetuses. AB - We report 2 cases of fetal heart failure associated with large placental chorioangiomas. One fetus exhibited serious hydrops on the initial fetal echocardiogram and was ultimately stillborn. The fetus in the other case exhibited cardiomegaly. Following the premature termination of the pregnancy, the fetus received medical treatment and recovered in 7 days. Monitoring the fetal cardiac size with ultrasonography is recommended to determine the optimal time of delivery in cases of large placental angioma that are diagnosed prenatally. PMID- 9354886 TI - Is inflammation on Papanicolaou smear a risk factor for preterm delivery? AB - OBJECTIVE: Inflammation on Papanicolaou (Pap) smear has been associated with a 30 50% incidence of bacterial vaginosis (BV), a recognized risk factor for preterm delivery. We determined whether inflammation on Pap smear is associated with preterm delivery. STUDY DESIGN: 5,348 cases were studied with complete prenatal data including the potential confounder of treatment with antibiotics. Cases were categorized by presence (n = 1,139) or absence (n = 4,209) of inflammation on Pap smear. RESULTS: In the inflammation group the proportion of African Americans was lower (66.9 vs. 74.5%; p < 0.001). There were no significant differences (t test) for maternal age, gravidity, history of preterm delivery, or gestational age at delivery between inflammation and noninflammation groups. Multiple stepwise regression analysis showed that maternal age, history of preterm delivery, and African American race were all significantly positively associated with preterm delivery. Treatment with metronidazole during the pregnancy was significantly negatively associated with preterm delivery. Inflammation on Pap smear, gonorrhea detected during the pregnancy and prenatal treatment with erythromycin were not associated. CONCLUSION: Unlike bacterial vaginosis, inflammation on routine Pap smear does not appear to be a risk for subsequent preterm birth. We are unable to use inflammation on Pap smear as a surrogate for more specific diagnosis of bacterial vaginosis. PMID- 9354887 TI - Meconium drug screening of stillborn infants: a feasibility study. AB - OBJECTIVE: Meconium drug testing of liveborn infants is highly sensitive (87%) and specific (100%). Accurate knowledge of drug use in mothers of stillborns would be beneficial. We determined the feasibility of noninvasive meconium drug screening for opiates and cocaine in stillborns. METHODS: Stillborn infants delivered at our hospital had meconium collected using a 4-mm spatula inserted into the anus. Specimens were analyzed using gas chromatography. Charts were reviewed. RESULTS: Of the 30 specimens obtained, 26 were below the optimal amount needed (0.5 g). Regardless, all samples were analyzed and three were positive for cocaine (10%), none for opiates. Two of the 3 positive samples were of 'insufficient quantity'. In one, the presumptive cause of fetal demise was diabetes, with no additional factors suggesting substance abuse. The other fetal loss was due to idiopathic preterm labor at 21.5 weeks, with a positive UDS. CONCLUSION: In this pilot study, inability to obtain an optimal volume of meconium occurred frequently. However, important and unexpected laboratory data were generated even with 'insufficient quantity'. This highlights the need to develop more refined methodologies for this screening tool in stillborn fetuses. PMID- 9354888 TI - Cordocentesis using the combined technique; needle guide-assisted and free-hand. AB - Two hundred and sixteen diagnostic cordocenteses were performed using the following technique: A guide was used to deliver the distal end of the needle to the immediate vicinity of the umbilical cord, after which the needle was released from the guide and a free-hand technique was used to enter the umbilical cord. The vessel punctured was identified by its sonographic appearance and flow direction using color Doppler technology. All procedure-related losses which occurred within 2 weeks were analyzed. The gestational age at the time of cordocentesis ranged from 18 to 42 weeks. Most punctures (62%) were performed at the placental insertion of the umbilical cord. In 32% of fetuses the free floating loop was sampled and in 6% the puncture was performed at the site of cord entry into the fetus. Two fetuses died shortly after cordocentesis. One death occurred at 28 weeks in a fetus with severe cytomegalovirus infection. The other death was due to premature rupture of the membranes after the procedure in a very premature fetus. The overall fetal loss rate was 0.93%. In conclusion, the combination of the two cordocentesis techniques appears safe and highly successful in obtaining fetal blood samples. PMID- 9354890 TI - Fluorescence in-situ hybridization (FISH) reveals that in chronic myelogenous leukaemia (CML) following interferon-alpha therapy, normalization of megakaryocyte size is associated with the loss of bcr/abl translocation. AB - AIMS: In addition to predominant granulocytic proliferation, bone marrow morphology in Philadelphia chromosome positive (Ph1+) CML is characterized by atypical dwarf or microforms of megakaryocytes. However, following therapy with interferon-alpha 2b (IFN), these micromegakaryocytes occur less frequently. The purpose of this study was to elucidate whether the reappearance of normal megakaryocytes may be associated also with a reduction of the bcr/abl-positive cell clone. METHODS AND RESULTS: Fluorescence in-situ hybridization (FISH) technique in combination with immunomorphometry (CD61) was performed on trephine biopsies. A total of 311 CD61-positive megakaryocytes, including precursors and atypical microforms, were evaluated in pre-treatment specimens derived from 11 patients with Ph1+ CML. A specific fusion site marking the bcr/abl translocation was found in 87% of megakaryocytes which showed a size of 169 +/- 35 microns2. In untreated patients, atypical microforms (size 200 microns2) were observed in 66% of the total megakaryocytic population. Following IFN therapy 369 megakaryocytes could be analysed in sequential examinations and were found to display a significant decrease (63%) in positive fusion signals. In addition there was also a significant enhancement in average size (252 +/- 66 microns2) reflecting a reduction in the number of micromegakaryocytes (43%). These findings were particularly conspicuous in three patients with a major to complete cytogenetic remission. CONCLUSIONS: A normalization of megakaryocyte size following IFN therapy in CML is significantly associated with a loss of the bcr/abl translocation site and therefore indicates a (partial) recovery of normal haematopoiesis. PMID- 9354891 TI - Lesions of 13q may occur independently of deletion of 16q in spindle cell/pleomorphic lipomas. AB - AIMS: Very recent multidisciplinary investigations have allowed for the definition among lipomas of a clinical and histological subtype called spindle cell and/or pleomorphic lipoma, possibly associated with partial monosomy 16 and anomalies of chromosome 13. In order to get nearer to the underlying critical molecular changes further multidisciplinary pathological and genetic research is indicated, to identify which chromosome(s) anomalies are crucial in the development of these tumours. METHODS AND RESULTS: In an ongoing multidisciplinary study of lipomatous tumours, including clinical findings, morphology, histochemistry and cytogenetics, two instances were found of spindle cell lipoma with clonal chromosome changes. In both cases chromosome 13 was involved, whereas only one showed a partial monosomy 16. CONCLUSIONS: Partial monosomy 16 is a characteristic lesion in spindle cell lipoma, usually associated with anomalies of chromosome 13. The present report confirming a previous single observation indicates, however, that lesions of 13 may occur independently from lesions of 16, suggesting different underlying molecular lesions in these otherwise very similar lipomas. PMID- 9354889 TI - Significance of metallothionein overexpression in human tumours. PMID- 9354892 TI - p53 tumour suppressor gene protein expression in early and advanced gallbladder carcinoma. AB - AIMS: Gallbladder carcinoma is one of the most frequent malignant tumours occurring in Chile and the mortality rate in both sexes ranks among one of the highest in the world. Mutation of p53 tumour suppressor gene has been demonstrated in many tumours. Our aim was to determine protein expression of p53 gene in early and advanced gallbladder carcinoma. METHODS AND RESULTS: Protein expression of gene p53 was studied by immunohistochemical means in 191 gallbladder carcinomas (157 primary tumours, 34 metastases) and 25 controls. In 86 out of 191 cases (45%), protein expression of gene p53 was observed. Differences related to sex, age, or race were not observed. All gallbladder controls were negative. Twenty-five per cent of well-differentiated tumours were p53 positive, while moderate or poorly differentiated carcinomas reached 50% (P = 0.04). p53 expression was observed in 23.5% of early carcinomas and in 48.2% of advanced carcinomas (P = 0.01). No differences between primary tumours and metastasis were demonstrated. CONCLUSIONS: Protein expression of p53 tumour suppressor gene is observed in 45% of gallbladder carcinomas. The absence of expression in controls and in normal mucosa adjacent to tumours suggests its utility in differentiating atypical gallbladder epithelia from neoplastic lesions. PMID- 9354893 TI - Thyroid carcinoma associated with familial adenomatous polyposis. AB - AIMS: Thyroid carcinoma is an extracolonic manifestation that is present in about 1% to 2% of patients with familial adenomatous polyposis (FAP). Less than 100 cases have been reported in detail. We have investigated the suggestion that FAP associated thyroid carcinoma is significantly different morphologically from both papillary and follicular types and can be considered as a separate entity. METHODS AND RESULTS: Specimens from three patients with FAP associated thyroid tumours, all but one having single nodules, have been analysed. All three patients belonged to an extended kindred (23 siblings in four generations) who had genetic analysis and intensive screening for thyroid nodules. Seven patients had the same APC mutation at codon 1061. Pathological examination revealed a typical papillary carcinoma, encapsulated variant, in all patients, with follicular areas in one case. All thyroid specimens, in addition to histological and immunohistological examinations, were also specifically studied for activation of the RET-PTC oncogene, that seems to be restricted to papillary thyroid carcinoma. Two of the three patients had RET-PTC activation (PTC1 isoform). CONCLUSIONS: The findings suggest that the tumours were certainly papillary, at least in the present kindred. Further studies in different families are required for a better understanding of this peculiar tumour and of its biological behaviour. PMID- 9354894 TI - Role of microliths in the aetiology of chronic submandibular sialadenitis: a clinicopathological investigation of 154 cases. AB - AIMS: Confusion about the aetiology and pathogenesis of chronic submandibular sialadenitis led to the present investigation of 154 cases in which many clinical and histological features were analysed. METHODS AND RESULTS: By far the greatest number of histological factors, namely liths, atrophy, fibrosis, parenchymal inflammation, lymphoid germinal centres, mucous and ciliary metaplasia, salivary extravasation and glycosaminoglycan accumulation, was related to the degree of inflammation, which appears to be of the greatest importance in the aetiology and pathogenesis. Inflammation, atrophy and fibrosis were related to duration of symptoms, which supports the concept of a chronological progression through increasingly severe histological changes. CONCLUSIONS: Inflammation possibly arises from ascending infection in a normal gland and exerts an obstructive and destructive effect on the parenchyma with the development of the related histological changes and a vicious circle involving further ascending infection. Normal glands contain microliths that possibly by localized obstruction cause atrophic foci that are reservoirs for ascending infection. Microliths and liths were unrelated: microliths were related to age as in normal glands whereas liths were related to duration of symptoms and appeared to be secondary to the sialadenitis. Many glands showed minimal changes, which raises the possibility of conservative treatment. PMID- 9354895 TI - Systemic-to-pulmonary vascular malformation of lung visualized by computer assisted 3-D reconstruction. AB - AIMS: While a growing number of cases with pulmonary arteriovenous malformation (PAVM) have been reported, detailed analysis has yet to be found on the relation of abnormal vessels with the whole lung vasculature and airways. To gain more insight into the structure-function interrelation of this disease, we attempted to visualize the vessels in and around the arteriovenous malformation, resorting to computer-aided 3-D reconstruction. METHODS AND RESULTS: The material was the upper lobe of the right lung from a 44-year-old man resected for recurrent haemoptysis. On pre-surgical selective angiography, an arteriovenous communication was suggested to exist between a tributary of the right 3rd intercostal artery and pulmonary vein. Semi-serial sections were prepared from the material and submitted to 3-D reconstruction of blood vessels and airways. In 3-D images, branches of the 3rd intercostal artery proved to be forming a plexus of abnormally dilated, thin-walled vessels in the subepithelial layer of a membranous bronchiole, a situation clearly explaining the mechanism of haemoptysis. There was no capillary bed interventing between the afferent arteries and draining vessels leading to the pulmonary vein. CONCLUSIONS: This presents the first overall visualization of PAVM, allowing comparison of 2-D microscopy with the corresponding 3-D morphology. PMID- 9354896 TI - Granular cell dermatofibroma. AB - AIMS: To describe a series of five granular cell dermatofibromas as an unusual and rare manifestation of fibrohistiocytic tissue response. METHODS AND RESULTS: Five granular cell dermatofibromas were collected out of 136 tumours filed as granular cell tumours. Clinically, all lesions occurred on the shoulder or back of middle-aged adults (two women, three men), mostly with the clinical diagnosis of a fibrohistiocytic lesion. Histology revealed well-circumscribed, dermal to subcutaneous lesions dominated by periodic acid-Schiff (PAS) positive, granular cells. Acanthosis above, as well as storiform arrangement of spindle cells, sclerotic collagen and some interspersed lymphohistiocytic infiltrate at the periphery of the lesion, indicated the fibrohistiocytic origin. Lesions showed prominent reactivity with NK1C3 (CD57), as well as for macrophage markers KiM1p and KP1 (CD68). In contrast to classic Schwannian/neurogenic granular cell tumours, granular cell dermatofibromas were S100 protein negative, but showed variable reactivity for factor XIIIa (10-50%) in 4/5, for smooth muscle specific actin (10-50%) in 2/5 and with E9 (10-30%) in 3/5 lesions. Electron microscopy in one case revealed large pools of phago-lysosomes and variably sized glycogen granules in granular cells. CONCLUSION: Our series delineates granular cell dermatofibroma as a distinct clinicopathological variant of fibrohistiocytic tissue response which needs to be distinguished from other tumours with granular cell features. PMID- 9354897 TI - Carcinosarcoma of the oesophagus showing neuroendocrine, squamous and glandular differentiation. AB - AIM: A case of oesophageal carcinosarcoma occurring in a previously fit, 64-year old man is reported. CASE SUMMARY: The carcinomatous component displayed neuroendocrine, squamous and glandular differentiation: the sarcomatous component showed no specific features of differentiation. In-situ squamous carcinoma was present in the adjacent squamous mucosa. The most superficial part of invasive tumour consisted of carcinosarcoma with a predominant neuroendocrine epithelial component. Squamous carcinoma without an accompanying sarcomatous component occupied most of the deeper part of the tumour, suggesting outgrowth of this tumour type by a selective growth advantage. CONCLUSION: We speculate that further tumour growth might have led to complete replacement of the tumour by pure squamous carcinoma, and that other advanced oesophageal squamous carcinomas might have had their origin in a short-lived carcinosarcomatous phase. PMID- 9354898 TI - Prognostic value of high serum levels of CA-125 in malignant secretory peritoneal mesotheliomas affecting young women. A case report with differential diagnosis and review of the literature. AB - AIMS: Very few cases of diffuse, malignant, peritoneal mesothelioma have been reported in young women. Distinction between peritoneal mesothelioma and serous epithelial tumours, including papillary serous carcinomas and borderline serous tumours, can be difficult. Differential diagnosis based on clinical appearance and imaging techniques is broad and inconclusive, thus the diagnosis must be confirmed by histological examination. Because the vast majority of tumours involving the peritoneal and serosal surfaces are due to primary or metastatic serous epithelial tumours, there is a tendency on part of pathologists to disregard the possibility of mesothelioma when examining a biopsy or excision specimen. This is especially likely to occur when mesothelioma is associated with highly elevated serum levels of CA-125, which is the typical tumoral marker of epithelial serous tumours from the ovary. The association between peritoneal mesothelioma and high serum levels of CA-125 has been reported in the literature only in two cases. CASE DETAILS: In order to avoid a misdiagnosis of this neoplasm we describe a new case of peritoneal mesothelioma in an 18-year-old woman with high serum levels of CA-125. CONCLUSIONS: Besides its clinicopathological characteristics and its histological, immunohistochemical and ultrastructural features, we describe its biological behaviour, which seems to be worst when CA-125 levels are high. PMID- 9354899 TI - Large cell neuroendocrine carcinoma of the thymus. AB - AIM: We highlight the occurrence of an unusual neuroendocrine tumour, a large cell neuroendocrine carcinoma, arising from the thymus. CASE DETAILS: A 68-year old man with a history of cigarette smoking had a large mediastinal tumour arising from the thymus removed. Two years later the tumour recurred; it was debulked surgically but the patient died 2 months later: Histological examination of both tumour specimens revealed a tumour with an endocrine pattern, composed of large pleomorphic cells with large nuclei and prominent nucleoli. The mitotic count ranged from 19 to 26 per 10 high-power fields and large tracks of coagulative tumour necrosis were present. The tumour cells were strongly positive for neuron-specific enolase (NSE), chromogranin, CAM5.2 and AE1/3, with cytoplasmic dot-like accentuation for the latter three markers. The tumour fulfilled the criteria for a diagnosis of large cell neuroendocrine carcinoma. CONCLUSIONS: Large cell neuroendocrine carcinoma should be distinguished from atypical carcinoid and small cell carcinoma. It is a distinctive neuroendocrine malignancy with a prognosis between that of atypical carcinoid and small cell carcinoma, and needs to be treated aggressively. PMID- 9354900 TI - Intestinal metaplasia subtyping: evaluation of Gomori's aldehyde fuchsin for routine diagnostic use. AB - AIMS: Intestinal metaplasia (IM) has been implicated in the pathogenesis of gastro-oesophageal carcinoma, but because of its common occurrence, its specificity for use in cancer surveillance is low. IM subtypes characterized by mucin phenotype have been studied to try and improve specificity. METHODS AND RESULTS: On balance, type III IM seems the most promising for use in gastric cancer surveillance. The situation is problematic at the gastro-oesophageal junction where the normal occurrence of acidic mucins raises doubt on the value of subtyping. High iron diamine-Alcian blue combination (HID-AB) is commonly used for IM subtyping, but its potential toxicity and long staining period (up to 24 hours) precludes widespread clinical use. This study has compared the sulphomucin staining ability of Gomori's aldehyde fuchsin-Alcian blue combination (GAF-AB) against HID-AB for identifying and subtyping IM in gastric and oesophageal biopsies. CONCLUSIONS: Compared to HID-AB, a sensitivity of 85%, a specificity of 100% and a staining time of less than 30 minutes, shows this stain to be a simple and effective technique for identifying and subtyping IM in routine laboratories. PMID- 9354901 TI - Rapid verification of the identity of questionable specimens using immunohistochemistry with monoclonal antibodies directed against HLA-class I antigens. AB - AIMS: A case report is presented in which an unexpected pathological diagnosis raised the possibility that biopsies of two patients were mixed-up. Since these biopsies were obtained from kidney transplant patients, the HLA-typings of both patients were known. METHODS AND RESULTS: We developed an immunohistochemical method using HLA-class I specific monoclonal antibodies to recognize the donor and recipient antigens in these biopsies. Using this method we could confirm the identity of the patients of whom the biopsies had been taken. CONCLUSIONS: This method, which uses the highly polymorphic HLA-system, is potentially useful for rapid and easy verification of the identity of specimens if a mix-up is suspected. PMID- 9354902 TI - Malignant struma ovarii' with peritoneal dissemination. PMID- 9354903 TI - Oncocytic metaplasia in a adenoma of the gallbladder. PMID- 9354904 TI - Spitz naevus with a predominant epithelioid cell component and halo reaction. PMID- 9354905 TI - Osteogenic melanoma: stromal metaplasia in association with subungual melanoma. PMID- 9354906 TI - Microscopic, lymphocytic and collagenous colitis. PMID- 9354907 TI - Application of biomarkers in cancer epidemiology. Workshop report. AB - In epidemiology, a biological marker (commonly abbreviated, for convenience, to biomarker) is any substance, structure or process that can be measured in the human body or its products and may influence or predict the incidence or outcome of disease. Biomarkers can be broadly classified into markers of exposure, effect and susceptibility. Biomarkers may include the following: xenobiotic agents and their metabolites in tissues or body products; normally occurring body constituents whether in physiological or pathological amounts; endogenous compounds that are not present under normal conditions; and inherited and acquired abnormalities of body chemistry, structure or function, including pathological manifestations of precursors to disease. Biomarkers should be distinguished from biomarker assays, specific laboratory tests aimed at measuring particular biomarkers, and biomarker measurements, the amounts of particular biomarkers present in specified units of tissues or body products as measured by biomarker assays. PMID- 9354908 TI - Transitional studies. AB - Transitional studies are studies using biological markers that bridge the gap between laboratory experiments and population-based epidemiology. The goal of these studies is to characterize and validate biomarkers and to assess the following: intra- and inter-subject variability; the feasibility of marker use in field conditions; confounding and effect-modifying factors for the marker; and mechanisms reflected by the biomarker. Another goal is to optimize the conditions for the use of biomarkers. Transitional studies involving biomarkers of exposure or effect are distinguished from etiological studies because the biomarker is generally the outcome or dependent variable. Despite this difference, transitional studies can be epidemiological studies, but they may also include laboratory studies to assess reliability (and accuracy) and to identify parameters for collecting, processing and storing biological specimens prior to assay. Generally, transitional studies involve healthy people, patients or workers with specific exposures. At some point in the validation of a biomarker the line between transitional and etiological studies becomes blurred. None the less, it is useful to identify transitional studies as a distinct set of efforts to validate and characterize biomarkers. Transitional studies can be divided into three functional categories: developmental, characterization and applied studies. PMID- 9354909 TI - Logistics and design issues in the use of biological samples in observational epidemiology. AB - Standard epidemiological study designs are well suited to answering questions involving the collection of biological samples. However, different designs are better suited--both for design and logistic reasons--to different questions. The strengths and weaknesses of each design are discussed in relation to markers of exposure, susceptibility and early outcome, and to markers used to classify cancers into biologically defined subsets. PMID- 9354910 TI - Methodological issues in the use of biological markers in cancer epidemiology: cohort studies. AB - In this chapter we summarize the major strengths and weaknesses of cohort studies; consider how these characteristics influence the use of biomarkers in cohort studies; briefly review considerations of statistical power, design and the influence of measurement error in cohort studies; and discuss some of the emerging ethical considerations that relate to the use of biomarkers in prospective studies. PMID- 9354911 TI - General issues of study design and analysis in the use of biomarkers in cancer epidemiology. AB - Other contributions to this volume have discussed sources of variation (see Vineis) and measurement error (see White). In this article, we focus on statistical issues involved in the design and analysis of epidemiological studies that use biomarkers. We do not consider statistical issues of laboratory analyses. PMID- 9354912 TI - Sources of variation in biomarkers. AB - Epidemiology is interested in variation. The goal of epidemiological research is to infer cause-effect relationships by observing whether the occurrence of disease varies according to relevant exposures. Epidemiology is usually interested in 'intergroup' variation (e.g. between those who are exposed and those who are unexposed to the factor of interest), while 'intragroup' variation is a source of noise. Therefore, the study design aims to increase intergroup variability, to limit intragroup variability and to reduce error by ensuring validity and reliability of measurements. The goal of this chapter is to summarize simple ways to recognize the main sources of measurement error and to estimate the extent and the impact of such error. PMID- 9354913 TI - Effects of biomarker measurement error on epidemiological studies. AB - This chapter presents an overview of how measurement error in a biomarker affects epidemiological studies which use the biomarker. To estimate the effects of biomarker error, one must first measure the error using an appropriate validity or reliability study design and using appropriate parameters, i.e. parameters that are informative about the effects of measurement error on the 'parent' epidemiological study. These measures of the biomarker measurement error from the validity or reliability study can then be applied to what is known about the association under study in the parent study, in order to estimate the effects of the biomarker error on the results of the epidemiological study. PMID- 9354914 TI - Markers of internal dose: chemical agents. AB - Biomarkers of internal dose measure the level of a carcinogen or one of its metabolites in a tissue or a body fluid such as urine or blood. The choice of a biomarker of internal dose for a particular epidemiological study or type of study requires careful consideration of the period of exposure to which the biomarker relates, host factors related to carcinogen metabolism, invasiveness of sampling, reliability and cost of the biomarker. Before a new biomarker is adopted, it is important to assess these characteristics in transitional studies to ensure that the biomarker will be applied appropriately. Biomarkers of internal dose have been applied most successfully in ecological studies and nested case-control studies, and are especially useful when they provide information about long-term carcinogen exposure. PMID- 9354915 TI - Biochemical markers of dietary intake. AB - The primary objective of nutritional epidemiology is to identify, in combination with other forms of research, which aspects of diet and nutritional factors are causally related to cancer development. However, traditional epidemiology can evaluate with only a limited degree of specificity to which individual dietary factors an increased occurrence of cancer can be attributed. The two main reasons for this are: (1) dietary intake levels of specific foods or food constituents can be strongly intercorrelated; (2) dietary intake levels of specific food constituents are generally measured with rather large errors. Biochemical markers are increasingly seen as measurements that may help to overcome some of the above mentioned methodological problems in nutritional epidemiology. PMID- 9354917 TI - Carcinogen-DNA and carcinogen-protein adducts in molecular epidemiology. AB - Carcinogen-DNA and carcinogen-protein adducts provide an integrated measure of carcinogen exposure, uptake and absorption, metabolism, DNA repair and cell turnover. As such they promise to provide a more objective and relevant measure of exposure than that which can be derived from questionnaires and measures of ambient levels of carcinogen. Nevertheless, the interpretation of adduct measurements made in human tissues and body fluids requires an understanding of a number of factors. These include the sensitivity and specificity of the measurement, the temporal relationship between exposure and adduct level and the mechanistic role of the adduct in the process of carcinogenesis. The application of such biomarkers in epidemiological studies therefore necessitates careful consideration of optimal study design. The above issues are illustrated in this chapter with examples from studies in both animal models and human populations. PMID- 9354916 TI - Biomarkers for biological agents. AB - Biomarkers of exposure to biological agents have proved to be extremely useful in establishing causal associations between infections and human cancer, as well as in tracing the natural history of the relevant agents. This chapter will focus on biomarkers for biological agents currently recognized as causally associated with various human cancers after evaluation at the IARC. These are human papillomavirus (HPV), hepatitis B (HBV) and C virus (HCV) and Helicobacter pylori. A prototype of nucleic-acid-based biomarkers is detection of HPV DNA. It measures the presence of type-specific DNA at a given point in time. PCR-based assays are considered the method of choice for epidemiological investigations. The test requires collection of exfoliated cells or biopsies. Specimens can be kept stored at -20 degrees C for long periods of time. DNA degradation is low. Some of the limitations of the marker for cancer epidemiology lie in the fact that HPV DNA infections are often transient, especially in young women. In repeated measurements, HPV DNA may fluctuate, but the reasons for this are unknown. Antigens and antibodies from the HBV and HCV can be viewed as prototypes for serological biomarkers. For HBV, there are markers able to distinguish between past and persistent infections. HBV DNA detection in sera further refines the assessment of exposure. Standardized serological assays are available and widely used in developing countries. For HCV antibodies, serological assays are standardized and widely available. RNA detection in sera by PCR is under development. A limitation of the currently available assays is the type and subtype variation of HCV by geography, which requires further research and standardization. In low-risk populations, numerous false-positive results arise and confirmatory tests are required. PMID- 9354918 TI - Somatic cell mutations in cancer epidemiology. AB - Somatic cell mutations arising in vivo in reporter genes and in cancer-associated genes may now be measured in humans. Background mutation levels and mutational responses following various mutagen exposures are reviewed in this chapter. The detection methods are compared for similarities and differences based on the underlying biology of the systems. Currently available data on molecular mutational spectra are reviewed and the utility of such information is discussed in terms of mutagen exposure characterization and for defining the mutagenic basis of carcinogenesis. In addition to the reporter gene assays, recently developed assays for mutation in cancer-associated genes are considered. The strengths and limitations of using somatic cell mutations for cancer epidemiology and areas for future research are discussed. PMID- 9354919 TI - Cytogenetic end-points as biological dosimeters and predictors of risk in epidemiological studies. AB - Cytogenetic end-points have been successfully used in epidemiological studies for many years. Conventional end-points are now being replaced by procedures that utilize molecular methods, with greatly increased sensitivity, specificity and precision. In this paper we briefly review the most common cytogenetic assays that are useful in epidemiological settings, including structural chromosome aberrations, micronuclei, sister chromatid exchanges and analysis of interphase cells for aneuploidy. We describe new developments of each assay, where applicable, and discuss the strengths and weaknesses of the assays for detecting exposures and estimating risks. Finally, pertinent information concerning each of the assays that is useful in designing epidemiological studies is summarized in a table. It is hoped that the information presented here will be useful to individuals who are interested in applying biomarkers to studies of human environmental exposure and disease. PMID- 9354921 TI - Quality control of biomarker measurement in epidemiology. AB - The throughput and complexity of a biomarker assay will determine the amount of effort that can be expended on quality control and assurance. Clinical chemistry quality control procedures can be readily applied to simpler chemical analysis such as blood lead and cholesterol, but even complex cell-based biomarker techniques such as HPRT mutation analysis and cytogenetics benefit from a formal quality control approach. Collaborative interlaboratory exercises are essential, especially when no certified reference material is available, and these can play a central role in the control of laboratory drift. Recommendations are made for the quality control of biomarker measurement based on clinical chemistry techniques. These include recommendations for coding samples so that the laboratory scientist is unaware of exposure status and for the use of formal laboratory protocols. PMID- 9354920 TI - Methodological issues in the use of tumour markers in cancer epidemiology. AB - In this chapter, we review major methodological and practical issues associated with the use of tumour markers. At this stage of development, studies with a combination of tumour, susceptibility and exposure markers are needed to illustrate the link between exposure and biological response and to assess the interactive effects of tumour susceptibility markers in this process. Several practical issues related to the application of tumour markers are discussed, including banking of tumour tissue, setting a laboratory strategy and performing etiological heterogeneity analysis. PMID- 9354922 TI - Sample collection, processing and storage. AB - We review issues related to the inclusion of biospecimens in epidemiological studies. Technical advances and the revolution in molecular biology have rendered the use of biomarkers increasingly feasible in epidemiological investigations, however the cost and complexity require interdisciplinary expertise and careful attention to methodological detail in order to ensure validity. The widespread banking of biospecimens for long-term (cohort) studies requires special attention to be paid to these issues. Blood, urine and tumour tissue are in common use in medicine and at least some aspects of sample handling derives directly from this clinical experience, although special considerations apply in the epidemiological setting. An increasingly broad array of biospecimen types have been studied, including exhaled air, nail clippings, buccal cells, saliva, semen, faeces and breast milk. Relevant issues in the processing, storage, shipping, timing of collection and safety procedures are examined in terms of their potential to distort results. The role of carefully developed quality control protocols is emphasized. In order to take full advantage of the opportunities afforded by the use of biomarkers in epidemiological studies, careful attention to biospecimen processing, the stability of the biomarker and the precautions to be taken during transportation and storage of samples is necessary. PMID- 9354923 TI - Issues involving biomarkers in the study of the genetics of human cancer. AB - The investigation of hereditary factors in human cancer was suggested from kindreds that exhibited aggregations of cancer consistent with Mendelian inheritance. A subset of cancer that exhibits strong familial tendencies is due to single genes that 'cause' cancer; more commonly, hereditary factors may influence tumorigenesis in a stepwise probabilistic rather than deterministic manner through a variety of mechanisms, e.g. influencing the disposition of carcinogens. The roles of both common susceptibility genes and rare 'familial' cancer genes are receiving increasing attention in the general population. Population-based studies designed to examine more common genetic variants differ from linkage-based studies. Candidate susceptibility genes may be studied by phenotype or genotype approaches, and the relative advantages and disadvantages of each approach are considered. The issue of gene-environment interaction, implicit in the concept of susceptibility genes, is considered. The influence of genetic factors on individual and attributable risk is addressed. PMID- 9354924 TI - Gene--environment interactions in the application of biomarkers of cancer susceptibility in epidemiology. AB - Metabolic susceptibility genes are important determinants of individual susceptibility to the effects of environmental carcinogens. These genes follow the form of 'type 2' gene-environment interaction, whereby the polymorphic genetic risk factor functions only in the presence of an environmental exposure. Two different effects of carcinogen dose have been observed for these genes. Sometimes, increasing dose leads to a decreasing interaction, so that cases with the genetic risk factor have lower exposures than those cases without it. Other examples of a direct dose effect, whereby increasing exposure leads to increased interaction, have also been described. We propose a model based on multiple logistic regression to assess the nature of the dose effect in this type of gene environment interaction. This model allows for distinction between these two dose effects, and other effects such as protective or non-interactive effects of environmental and genetic risk factors. PMID- 9354925 TI - Using and interpreting surrogate end-points in cancer research. AB - Researchers have proposed a broad range of molecular, cellular and histological markers as surrogate end-points for cancer (SECs). The effect of an intervention on a 'valid' SEC is concordant with its effect on cancer incidence. The validity of a potential SEC is determined primarily by the extent to which the marker is a necessary event on the causal pathway to cancer. Colorectal adenomatous polyp formation is an example of a reasonably valid SEC because these lesions are obligate precursors of most large bowel malignancies. However, the existence of a plausible major alternative causal pathway--one bypassing the potential SEC- weakens inferences from that marker to cancer. Moreover, unless the pathway to cancer operates nearly exclusively through the SEC, an SEC that is valid for one intervention or exposure may not be valid for another. Metabolic, ecological, observational epidemiological and intervention studies may yield data that are useful in revealing these causal interrelations of intervention (exposure), SEC and cancer. Empirical studies of three questions are pertinent: (1) What is the relation of the SEC to cancer? (2) What is the relation of the intervention (exposure) to the SEC? (3) To what extent does the SEC mediate the relation between the intervention (exposure) and cancer? Data on SEC measurement error are important in ascertaining the extent to which marker results have been attenuated by such error. It is essential to carry out these studies to evaluate potential SECs (such as epithelial cell hyperproliferation) with plausible major alternative pathways to cancer. At the present time, definitive evidence on etiology and prevention will emerge only from studies with cancer end-points or SECs that are, by and large, necessary steps on the causal pathway to malignant disease. PMID- 9354926 TI - Biomarker end-points in cancer chemoprevention trials. AB - Over the last decade, the Chemoprevention Branch, Division of Cancer Prevention and Control, National Cancer Institute, USA, has been developing drugs that will slow or stop the progression to invasive cancer of precancerous (pre-invasive) lesions generally termed 'intraepithelial dysplasia' or 'dysplasia'. Over 40 short-term clinical trials are in progress, testing the following classes of agents on precancerous lesions in the different major organ systems: antimutagens (N-acetylcysteine, oltipraz), antiproliferatives (difluoromethylornithine, dehydroepiandrosterone, selenomethionine), antioxidants (vitamin E, curcumin), anti-inflammatories (aspirin, piroxicam, ibuprofen, sulindac sulfone) and hormonally active agents (tamoxifen in breast ductal carcinoma in situ and finasteride in prostatic intraepithelial neoplasia). Because of the strong practical need to keep so many clinical trials as short-term as possible, certain tissue changes known to be associated with high cancer risk were selected for use as biomarker end-points in the trials, such changes being quantitatively assayed by computer-assisted image analysis. These 'surrogate end-point biomarkers' (SEBs) are based on the individual cellular morphological and functional changes universally used by histopathologists to diagnose the lesion of intraepithelial neoplasia (Riddell, 1984; Boone et al., 1992; Wright et al., 1994). High grades of this lesion precede invasive cancer in the great majority of cases, and therefore SEBs based on them are linked to high cancer risk. Table 1 summarizes some of the short-term clinical trials now being monitored by the Chemoprevention Branch. The SEBs abbreviated 'PPNN' in the figure are: proliferative index (P); ploidy (DNA histogram) (P); nuclear morphometry and chromatin texture (N); and nucleolar size and frequency (N). Computer-assisted image analysis is used to assay these features quantitatively, which gives the SEBs increased objectivity, reproducibility and sensitivity. Further details concerned with cancer chemoprevention trials using SEBs, and their relation to the field of cancer epidemiology, are given below. PMID- 9354927 TI - The use of biological markers as predictive early-outcome measures in epidemiological research. AB - One of the possible uses of biomarkers in epidemiological research is as early outcome measures to predict the occurrence of clinical disease and to elucidate the biological mechanism of pathogenesis. This use is conceptually less straightforward than the well established use of biomarkers to improve or extend exposure assessment or to study interindividual variations in disease susceptibility. In principle, this form of use could accelerate or otherwise facilitate etiological research. However, in practice, the recent review literature suggests that this mode of biomarker use, especially in cancer epidemiology, is the least clear-cut and the least well developed. The recurrent problem is identifying biomarkers that: (1) are on the causal pathway, (2) have a high probability of progression to clinical disease, and (3) account for all or most of the cases of the specified clinical outcome. Such biomarkers would be most useful if they conferred a long lead-time relative to clinical disease occurrence. PMID- 9354928 TI - The use of biomarkers to study pathogenesis and mechanisms of cancer: oesophagus and skin cancer as models. AB - Recent advances in molecular biology have made it possible to use genetic alterations associated with cancer as biomarkers to study the pathogenesis and mechanisms of cancer. However, the lessons that can be drawn from the analysis of alterations in a particular cancer gene are extremely dependent upon the biological context in which they arise. In this article, we discuss the biological significance of alterations in the p53 tumour suppressor gene in cancers of the oesophagus and of the skin. In both tissues, different forms of cancer occur at high frequency (squamous-cell carcinoma and adenocarcinoma in the oesophagus; squamous-cell carcinoma, basal-cell carcinoma and melanoma in the skin). We show that specific patterns of p53 alteration occur in these various cancers and that analysis of these alterations is useful to make inferences about the etiopathogenesis of cancers of the oesophagus and of the skin. PMID- 9354929 TI - Comparing measurements of biomarkers with other measurements of exposure. AB - The issue of the relative merit of biomarkers and alternative measures of exposure arises most commonly in the context of epidemiological studies aimed at hazard detection and quantification. When exposures are from biological agents, biomarkers are usually the first and often the only justifiable choice. In general, however, the relative merit of different types of exposure measurements need to be evaluated on a case-by-case basis. Biomarkers may be affected by random errors, time-related sampling errors, physiological confounding and disease-induced differential error, all of which need to be explicitly evaluated before embarking on the use of a biomarker in a full-scale epidemiological study. Random errors affecting biomarkers may be reduced by replication or combination of measurements, or both. Alternative measurements of exposure can be evaluated against a biomarker when there is adequate evidence for regarding the marker as the true measure of a biologically relevant exposure. PMID- 9354930 TI - Ethical and social issues in the use of biomarkers in epidemiological research. AB - The use of biomarkers in epidemiological research may raise ethical and social issues. These issues stem from the belief that research participants have 'rights' to appropriate information before, during and after studies so that they can make informed decisions. Ethical issues can arise during protocol development, obtaining participation, and in the interpretation and notification of text and study results. Additionally, there are ethical considerations concerning the use of biological specimens collected and stored for one purpose and subsequently used for other research purposes. A major ethical issue is the maintenance of participants' privacy and the confidentiality of their test and study results. Ethics committees need to be well-informed about the scope, limitations and expectations of biomarker research in order to be able to respond to social and scientific developments in the use of biomarkers. PMID- 9354931 TI - Reduction of cellular cisplatin resistance by hyperthermia--a review. AB - Resistance to cisplatin (cDDP) is a major limitation to its clinical effectiveness. Review of literature data indicates that cDDP resistance is a multifactorial phenomenon. This provides an explanation why attempts to reverse or circumvent resistance using cDDP-analogues or combination therapy with modulators of specific resistance mechanisms have had limited success so far. It therefore provides a rationale to use hyperthermia, an agent with pleiotropic effects on cells, in trying to modulate cDDP resistance. In this review the effects of hyperthermia on cDDP cytotoxicity and resistance as well as underlying mechanisms are discussed. Hyperthermia is found to be a powerful modulator of cDDP cytotoxicity, both in sensitive and resistant cells. Relatively high heat doses (60 min 43 degrees C) seem to specifically interfere with cDDP resistance. The mechanism of interaction has not been fully elucidated so far, but seems to consist of multiple (simultaneous) effects on drug accumulation, adduct-formation and -repair. This may explain why hyperthermia seems to be so effective in increasing cDDP cytotoxicity, irrespective of the presence of resistance mechanisms. Therefore, the combination of hyperthermia and cDDP deserves further attention. PMID- 9354932 TI - An enhanced electrical impedance imaging algorithm for hyperthermia applications. AB - Electrical impedance imaging is a technique which is under investigation as a noninvasive method of tracking subsurface temperature distributions and/or associated cellular response during hyperthermia. In previous work, a finite element image reconstruction algorithm for converting surface potential distributions recorded at discrete electrode positions into spatial maps of conductivity values was developed. This paper reports on a series of significant improvements in the basic image reconstruction approach. Specifically, the ability to recover both the resistive and capacitive components of tissue electrical impedance have been incorporated. In addition, the image enhancement schemes of (1) total variation minimization, (2) dual meshing, and (3) spatial low-pass filtering, have been added. Through a series of simulation studies involving both phantom-like and clinically-relevant geometries having discrete regions and continuously-varying electrical property profiles, a significantly improved ability to recover spatial images of electrical properties in the impedance imaging context is demonstrated. The results show that the new algorithm is much more tolerant of measurement noise with levels up to 1% causing relatively modest degradations in image quality (compared to 0.1% which was needed previously in order to produce high quality images). The recovered electrical properties, themselves, both resistive and capacitive, are also found to be quantitative in value with errors in the 10-20% range occurring in the majority of cases, although deviations can reach 40% or more when noise levels as high as 10% are used. Temperature estimation simulations show that maximum temperature errors are significantly reduced (to approximately 2 degrees C relative to more than 10 degrees C in previous thermal simulations) with the new algorithm; however, temperature accuracies of better than 0.5 degree C on average are still found to be difficult to achieve with electrical impedance imaging even when the enhanced image reconstruction approach is used. PMID- 9354933 TI - Changes in muscle blood flow distribution during hyperthermia. AB - Blood flow is a critical parameter for obtaining satisfactory temperature distributions during clinical hyperthermia. This study examines the changes in blood flow distribution in normal porcine skeletal muscle before, during and after a period of regional microwave hyperthermia. The baseline blood flow distribution during general anaesthesia and after the insertion of the thermal probes was established independently in order to isolate the changes due to hyperthermia. General anaesthesia alone and thermocouple insertion during anesthesia had no significant effect on the muscle blood flow distribution. Regional microwave heating generated a non-uniform blood flow distribution which was a function of the tissue temperature distribution. Blood flow was greater in those tissues samples in which higher temperatures were recorded and less in those sampled further from the applicators peak SAR (Specific Absorption Rate). The increase in blood flow appears to be primarily a local phenomenon. Although muscle blood flow may be considered to be uniform prior to heating, this does not hold during hyperthermia treatment. Therefore, the non-uniform nature of the blood distribution during heating should be incorporated into any practical bioheat transfer model. PMID- 9354934 TI - Long-duration, mild whole body hyperthermia with cisplatin: tumour response and kinetics of apoptosis and necrosis in a metastatic rat mammary adenocarcinoma. AB - This study examines antitumour effect and induction of apoptosis and necrosis after treatment with long-duration, mild whole body hyperthermia (LL-WBH, 40.0 degrees C, for 6 h) in simultaneous combination with cisplatin (CDDP) on primary and metastatic tumour growth in a rat mammary adenocarcinoma. A significantly greater delay in primary mammary tumour growth was observed after treatment with LL-WBH + CDDP, compared to either modality alone (p < 0.05). LL-WBH alone caused a significant delay in spontaneous metastasis to the axillary lymph node (ALN) and LL-WBH + CDDP tended to further increase the delay in ALN metastasis. Survival was longest in rats receiving LL-WBH + CDDP, compared to other groups (p < 0.05). CDDP induced a peak of tumour apoptosis at 24 h after treatment beginning that was significantly greater than LL-WBH alone (p < 0.05). The peak of tumour apoptosis induced by LL-WBH + CDDP from 12 to 24 h was significantly greater than any other group (p < 0.01). These results suggest that the extent of treatment-induced apoptosis seems to correlate positively with antitumour response and the combination or LL-WBH with CDDP may lead to a promising adjuvant therapy for breast cancer. PMID- 9354936 TI - Oxidants differentially regulate the heat shock response. AB - Cells, animals, and humans respond to hyperthermia through the synthesis of a family of proteins termed heat shock proteins (HSPs). Because hyperthermic stress may also result in mitochondrial uncoupling and the generation of reactive oxygen species, we wondered whether oxidant stress was sufficient to increase cellular levels of HSP70. HSP70 was detected in cells heated or treated with menadione but not in those treated with hydrogen peroxide or xanthine/xanthine oxidase. We speculate that oxidant stress from menadione exposure is qualitatively different from exposure from hydrogen peroxide or xanthine/xanthine oxidase. PMID- 9354935 TI - Hsp40, a possible indicator for thermotolerance of murine tumour in vivo. AB - The relationship between Hsp40/Hsp70 synthesis and the development of thermotolerance was investigated using mouse squamous cell carcinoma in vivo. To examine the thermotolerance, tumours were heated at 44 degrees C for 30 min as conditioning heating. After various intervals they were heated again at 44 degrees C for 90 min as challenge heating. The tumour response to heat was evaluated by the growth delay. Thermotolerance rapidly developed with increasing interval and reached a maximum at 12 h interval. Subsequently, thermotolerance gradually decayed and almost disappeared at 120 h interval. Under this condition, synthesis of Hsp40/Hsp70 increased after conditioning heating, reached a maximum at 12 h interval, then gradually decreased thereafter within 120 h. The kinetics of accumulation and decay of both Hsp40 and Hsp70 were very similar. The extent of thermotolerance was well correlated with the relative amount of Hsp40/Hsp70. These results obtained in vivo were very similar to those in vitro (Kaneko et al. 1995). Our findings suggest that Hsp40 could be a useful indicator of the degree of thermotolerance in addition to Hsp70 in vivo as in vitro. PMID- 9354937 TI - Lack of thermal enhancement for taxanes in vitro. AB - The taxanes represent a new class of clinical chemotherapeutic agents. A series of in vitro studies were independently of each other initiated in two different institutes (Amsterdam and Madison) to test the hypothesis that hyperthermia might enhance the cytotoxicity of taxanes. Clonogenic capacity experiments (Amsterdam) included the exposure of R1- and SW 1573-cells to 1, 4, or 24 h of paclitaxel with heat 43 degrees C x 60 min in the last hour of drug treatment or at 24, 48 as well as 72 h post drug treatment. Survival assay experiments (Madison) included the exposure of L-929-cells to paclitaxel and docetaxel for 24 h with heat 41.8 degrees C x 60 min the first or last hour of drug treatment as well as 24 and 48 h post treatment. No thermal enhancement of cytotoxicity for the taxanes was observed in these human and murine cell lines, with congruent data in both institutes. In addition, high performance liquid chromatography studies at 41.8 degrees C and 43 degrees C demonstrated paclitaxel and docetaxel were heat stable. PMID- 9354938 TI - Effects of an inhibitor of protein kinases on the response to heat treatment in cultured mammalian cells. AB - Effects of H7, a protein kinase C inhibitor, on responses to hyperthermic treatment were investigated in relatively heat sensitive Chinese hamster V79 cells and resistant human glioma A7 cells. In V79 H7 (2-50 microM) enhanced cell killing of heat treatment of 42 or 44 degrees C. The magnitude of the heat sensitization was dependent on concentration and timing of H7 addition; addition of the inhibitor between 0 and 2 h before heat treatment was most effective. In A7 the inhibitor did not show such synergistic effect with heat treatment, but showed mere added toxicity. In split-heat experiments using V79 with addition of H7 (20 microM) before the initial heat treatment and thereon, the development of thermotolerance was partially inhibited. However, already thermotolerant cells were not sensitized when H7 was added before the test heat. In V79 there was a tendency for H7 to accelerate cell death and DNA ladder formation by heat. No significant change was detectable in HSP70 induction determined by Western analyses although H7 seemed to accelerate shifting of HSP70 out of nuclei back into cytoplasm. These results indicate that heat sensitizing effect of H7 may depend on cell type and that the effectiveness of H7 depends on timing of addition. PMID- 9354940 TI - Cytokine levels in adult patients with solid tumours undergoing whole body hyperthermia (WBH) PMID- 9354939 TI - Apoptosis induced by hyperthermia and verapamil in vitro in a human colon cancer cell line. AB - The aim of this study was to determine the mechanisms responsible for the growth inhibitory effect of hyperthermia and verapamil in human colon cancer cell line HT-29. Apoptotic cell death was verified by flow cytometry analysis. The effect of treatment with hyperthermia and verapamil on the expression of apoptosis associated proteins including Bcl-2, p53, bax, and c-Myc was studied by Western blot analysis. Changes in intracellular calcium homeostasis was analysed by fluorescence microscopy. The combination of 42 degrees C hyperthermia and verapamil caused a significant delay of human colon cancer cell proliferation as a result of apoptosis. Administration of these agents alone did not cause any cell inhibitory effect. Our experiments have shown that HT-29 cells constitutively express apoptosis-promoting proteins, such as Bax and c-Myc, while they fail to produce Bcl-2. Therefore, we hypothesize that HT-29 cells must have Bcl-2 independent pathways to protect cells against death-inducing signals. Also, apoptosis of HT-29 cells produced by hyperthermia in the presence of verapamil is a p53-independent process. Verapamil, when it did not act as a calcium channel blocker or inhibitor of release from intracellular storages under hyperthermic conditions, accelerated the increase of [Ca2+]i in HT-29 cells which resulted in programmed cell death (apoptosis). PMID- 9354941 TI - Lack of evidence for whole body hyperthermia-induced changes in MHC and non-MHC immunological function via cytokine induction. PMID- 9354942 TI - Current status of gene therapy for prostate and bladder cancer. PMID- 9354944 TI - Efficacy of continuous subcutaneous infusion therapy using interferon alpha and the possible prognostic indicator of TNF-alpha in renal cell carcinoma. AB - BACKGROUND: In an attempt to improve efficacy by escalating the dose and maintaining higher serum concentrations over a long period of time, this study examines continuous interferon alpha (IFN alpha) subcutaneous infusion therapy in patients with renal cell carcinoma (RCC). METHODS: Seven of 11 patients with RCC had evaluable metastatic lesions. A highly purified natural human IFN alpha was injected subcutaneously via an infuser pump for 5 consecutive days, followed by a 2-day rest period (25 million IU/week). The treatment was continued for a period of 15 weeks. Serum concentrations of IFN alpha, IL-1 alpha, IL-1 beta, TNF-alpha, and IFN gamma were measured at intervals throughout the study period. RESULTS: Two of the 7 patients with evaluable lesions achieved a partial response (overall response rate, 29%), while 1 achieved a partial response only to lung metastasis. These 3 cases were defined as responders. No difference was found in the concentration of serum IFN alpha between responders and nonresponders, however, a significantly higher concentration of serum TNF-alpha was observed in responders (P < 0.05, Mann-Whitney U test). Five cases (45%) had moderate to severe adverse effects, including depression (n = 1), eyeground hemorrhage (n = 2), and general fatigue (n = 2). CONCLUSION: Appropriate patient selection may be necessary for subcutaneous continuous infusion therapy for the treatment of metastatic RCC. Also, the serum concentration of TNF-alpha measured during the course of treatment reflected well on the outcome of IFN alpha therapy. PMID- 9354943 TI - Differential diagnosis of primary benign and malignant retroperitoneal tumors. AB - BACKGROUND: Clinical differential diagnosis between malignant and benign tumors is important in order to select a therapeutic strategy for a primary retroperitoneal tumor. METHODS: The clinical findings and radiological features of 25 patients with primary retroperitoneal tumors were retrospectively evaluated to find those signs that might contribute to the preoperative distinction between benign and malignant tumors. RESULTS: Of 25 primary retroperitoneal tumors, 15 were benign. This may reflect the increased number of incidentally found small benign tumors. There were significant associations between the presence of symptoms and malignancy (P < 0.05), between irregular margins on imaging and malignancy (P < 0.05) and between the absence of calcification and malignancy (P < 0.05). Malignant tumors were significantly larger than benign tumors (11.45 +/- 1.90 cm vs. 5.31 +/- 0.43 cm). A retroperitoneal tumor scoring system was developed to distinguish primary retroperitoneal benign tumors from their malignant counterparts based on the: 1) maximum diameter equal to or larger than 5.5 cm, 2) presence of symptoms, 3) absence of calcification, 4) presence of irregular margins, and 5) presence of cystic degeneration or necrosis. A significant correlation was found between the incidence of malignant tumors and the total retroperitoneal tumor score (P < 0.05). CONCLUSION: This study suggests that the size of tumor, the presence of symptoms, irregular margins, and the absence of calcification may be valuable predictors of primary retroperitoneal malignant tumor. PMID- 9354945 TI - Histologic characteristics of renal cell carcinomas with lymph node metastasis. AB - BACKGROUND: This study was conducted to determine if there are any specific histologic features that are associated with lymph node metastasis in renal cell carcinoma (RCC). METHODS: TNM classification, histologic grade, mean nuclear volume, cell type, and histologic architecture of the tumors were evaluated in 66 patients who had undergone nephrectomy and lymphadenectomy for RCC. In the 18 patients with positive lymph node metastasis, both primary lesions and metastatic lymph nodes were evaluated. RESULTS: Lymph node status was correlated with primary tumor stage, venous involvement, and distant metastasis. The tumor grade was higher, and the mean nuclear volume was larger, in both primary and metastatic lesions of RCCs with lymph node metastasis than in tumors with no metastasis. In primary lesions of RCCs with lymph node metastasis, clear cell, alveolar, or cystic patterns were observed less frequently, and granular or spindle/pleomorphic cells and papillary or solid patterns, were observed more frequently, as compared to those lesions without metastasis. Comparison between primary and metastatic lesions in individual patients revealed no significant difference in grade or mean nuclear volume. The development of new cell types or histologic architectures, which was not noted in the primary lesions, was also a rare event in the metastatic lesions. CONCLUSION: Several characteristic histologic features, which may reflect the increased metastatic potential of the tumor, were observed in both primary and metastatic lesions in cases of RCC with lymph node metastasis. No substantial difference in histologic features was observed between the primary or metastatic lesions of individual patients. PMID- 9354946 TI - Influence of intravesical bacillus Calmette-Guerin therapy on systemic immunological status. AB - BACKGROUND: Intravesical bacillus Calmette-Guerin (BCG) therapy for superficial bladder cancer induces obvious local immunological responses. The purpose of this study was to examine the systemic immunological influences induced by intravesical BCG therapy. METHODS: We measured peripheral blood lymphocyte subsets (PBLS) and monocyte counts in 30 patients with superficial bladder carcinoma by two-colored flow cytometry before, during, and after intravesical BCG treatment. Comparisons were made between 24 good treatment responders and 6 poor responders. RESULTS: From 3 to 12 months after the beginning of treatment, PBLS and monocyte counts changed, as evidenced by an increase of suppressor T cells and a decrease of helper T cells. The good responders had higher cell counts than the poor responders, with differences in cell counts between good and poor responders more marked 1 month after beginning treatment, particularly of natural killer cells, cytotoxic T cells, and inducer T cells. These differences disappeared 3 months after the onset of treatment. CONCLUSION: Intravesical BCG therapy caused a marked and persistent alteration of the systemic immunological status. PMID- 9354947 TI - Significance of prostate-specific antigen after radical prostatectomy. AB - BACKGROUND: Serum prostate-specific antigen (PSA) is expected to be undetectable after radical prostatectomy unless there is residual disease or disease progression. The aim of this study was to confirm the usefulness of serum PSA measurements in monitoring patients after radical prostatectomy. METHODS: We conducted a study of 50 patients who underwent radical prostatectomy for clinical stage T1-2 or small T3 prostate cancer, analyzed serum PSA levels before and after surgery and compared the pathological findings and clinical outcome. RESULTS: Postoperative PSA elevation (PSA failure) was noted in 13% of patients with organ-confined disease (OCD), 43% with positive surgical margins and/or seminal vesicle involvement (PSM/SVI), and 60% with positive lymph nodes (N+). Postoperative clinical failure was noted in 8 patients, and all were preceded by PSA failures. The 3-year PSA failure-free rates were 86.5%, 32.1%, and 40.7% and the 3-year clinical failure-free rates were 100%, 67.5%, and 73.5% in patients with OCD, PSM/SVI and N+, respectively. CONCLUSION: An elevation of serum PSA levels after radical prostatectomy was a sensitive indicator of persistent disease after surgery, and preceded clinical manifestations of disease progression. PMID- 9354948 TI - Usefulness of preoperative MRI diagnosis in prostatic adenocarcinoma. AB - BACKGROUND: We examined the reliability of an MRI diagnosis prior to radical prostatectomy for prostate cancer. METHODS: A radical prostatectomy was performed in 24 patients with prostate cancer. Resected specimens were fixed and 5 mm step sections vertical to the urethra were prepared to resemble MRI images. We compared this pathological map with the preoperative MRI diagnosis which included capsular or seminal vesicle invasion and tumor localization in the prostate. We defined a new criterion for the presence of capsular invasion as a chemical shift that occurred on the rectal side on T1-weighted images 5 minutes after gadolinium (Gd) enhancement and the periprostatic venous plexus was not serial. We also examined 4 diagnostic factors of tumor localization including a low-signal intensity area detected in the peripheral zone on T2-weighted images, the presence of an enhanced area on Gd-enhanced T1-weighted images, and a low T2 with either Gd-enhanced or nonenhanced T1-weighted images. RESULTS: The accuracy of a preoperative MRI diagnosis of capsular invasion was 16.7% using the conventional criteria, but 88.9% adding the new criterion. The accuracy of predicting seminal vesicle invasion was 63.2% in a group using a body surface coil compared to 75% in the group using an endorectal surface coil. The accuracy, positive predictive value, sensitivity and specificity of diagnosing tumor localization were 69%, 74.4%, 35.1%, and 91.8%, respectively. CONCLUSION: This new criterion proved superior for diagnosing capsular invasion in prostate cancer patients. Also, analysis of tumor localization in the peripheral zone demonstrated that cancer detection is increased if the low-signal intensity area is enhanced by Gd. PMID- 9354949 TI - Clinicopathological features of prostate cancer detected by transrectal ultrasonography-guided systematic six-sextant biopsy. AB - BACKGROUND: The objectives of this study were to compare the efficacy of 3 modalities (prostate-specific antigen (PSA) assay, digital rectal examination (DRE), and transrectal ultrasonography (TRUS)) in detecting prostate cancer which was pathologically confirmed by TRUS-guided systematic six-sextant biopsy, and to investigate the relationship between the number of positive cores and several clinicopathological parameters. METHODS: Between 1992 and 1994, 297 males (155 from a mass screening program and 142 identified as outpatients) with a mean age of 71 years, underwent examinations including PSA determination, DRE, TRUS and systematic six-sextant biopsy, and/or additional directed biopsy. RESULTS: Prostate cancer was detected in 93 men. The sensitivity level of the PSA assay was significantly higher (85%) than that of either DRE or TRUS. Patients with an abnormal DRE or TRUS, elevated PSA levels, and those in the T3-T4 category or with moderate to poorly-differentiated adenocarcinomas had more positive biopsy cores (P < 0.05). Also, the relationships of both the number of positive biopsy cores and tumor grade to bone metastasis were significant (P < 0.01). Of 209 hypoechoic areas identified by transrectal ultrasonography, 42% were cancerous, and of 427 isoechoic areas, 12% were cancerous. The percentage of positive biopsy cores with hypoechoic areas was 86% in the subjects with a PSA > 10 ng/mL, but low (9%) in subjects with a PSA < or = 4 ng/mL, and the percentage of negative biopsy cores with a normal TRUS was high (98%) in subjects with a PSA of < or = 4 ng/mL, but lower (67%) in subjects with a PSA > 10 ng/mL. CONCLUSION: The serum PSA assay was more useful than either DRE or TRUS in detecting prostate cancer. The percentage of bone metastasis increased concomitant with the number of positive biopsy cores, and the positive biopsy rate of hypoechoic areas positively correlated with the PSA level. PMID- 9354951 TI - Long-term efficacy of a self-retaining intraurethral catheter for the treatment of prostatic obstruction. AB - BACKGROUND: Transurethral prostatectomy is a standard treatment modality for infravesical obstruction with benign prostatic hyperplasia. However, some patients cannot undergo this procedure, so alternative devices or surgical procedures have recently been developed. METHODS: We treated 26 patients with prostatic outflow obstruction by placement of a self-retaining intraurethral catheter (IUC). All patients had coexisting medical complications which contraindicated transurethral prostatectomy. RESULTS: In the short-term, the treatment was successful in 96.2% of the patients, however, longer follow-up demonstrated a significant failure rate, and the 3-year success rate declined to 12.5%. The majority of late failures were associated with urinary tract infections. CONCLUSION: These results showed that although placement of an IUC to treat prostatic obstruction was effective in the short-term, its long-term efficacy was highly limited. PMID- 9354950 TI - Therapeutic efficacy of clenbuterol for urinary incontinence after radical prostatectomy. AB - BACKGROUND: Urinary incontinence is one of the most common complications occurring after radical prostatectomy. We evaluated the efficacy of clenbuterol, a selective beta 2-adrenoceptor agonist, in the treatment of incontinence occurring after radical prostatectomy, using urodynamic assessment. METHODS: Fourteen men (mean age, 68 years) with post-radical-prostatectomy incontinence were treated with 20 mg of clenbuterol twice a day for 1 month. The urodynamic assessment was performed on all patients before and after the administration of clenbuterol. A pad scoring system was used to gauge the severity of incontinence before and after treatment. RESULTS: At 1 month after administration of clenbuterol, 9 of the 14 patients (64%) had dramatic improvement in pad scores. Treatment failed in 5 patients (36%) with severe incontinence. The results of urodynamic studies showed that the mean functional urethral length of the patients with post-radical-prostatectomy incontinence increased significantly after treatment. CONCLUSION: These results suggest that clenbuterol can be used as an effective agent for treating mild-to-moderate stress incontinence after radical prostatectomy. PMID- 9354952 TI - Nerve-sparing retroperitoneal lymph node dissection for metastatic testicular cancer. AB - BACKGROUND: Nerve-sparing techniques are used during retroperitoneal lymph node dissection (RPLND) in patients with early stage testicular cancer to preserve postoperative ejaculatory function. Indications for the procedures have been extended to patients with a postchemotherapy retroperitoneal residual mass without compromising the efficacy of surgery. We report 6 cases diagnosed with metastatic testicular cancer who underwent nerve-sparing RPLND. METHODS: Between January 1994 and March 1996, 6 patients with metastatic testicular cancer underwent nerve-sparing RPLND. Five of these patients received primary chemotherapy and a retroperitoneal residual mass. Four patients underwent complete bilateral RPLND and 2 underwent unilateral template surgery. RESULTS: After a mean follow-up of 18.7 months (range, 8 to 34), there have been no local recurrences and 5 (83%) patients report antegrade ejaculation. CONCLUSION: Nerve sparing RPLND is applicable for selected patients with metastatic testicular cancer without increasing the risk of local recurrence. Ejaculatory function is preserved in the majority of patients, contributing to the improvement of the quality of life in men who require such surgery. PMID- 9354953 TI - Internal iliac arterial infusion chemotherapy for rabbit invasive bladder cancer. AB - BACKGROUND: Internal iliac arterial infusion (IA) chemotherapy has been used clinically for locally invasive bladder cancer, but there have been no experimental studies to actually demonstrate whether IA is more effective than intravenous infusion (i.v.) chemotherapy in this setting. METHODS: We compared the effects of IA and i.v. using a rabbit invasive bladder cancer model. A 0.2 mL suspension containing 2 x 10(6) VX2 cancer cells was inoculated into the posterior submucosa of the bladder. Two weeks later the rabbits were divided into 3 treatment groups of 8 rabbits each: controls, a group treated with IA consisting of 10 mg/kg carboplatin and 1 mg/kg pirarubicin once a week for 3 weeks (days 14, 21, and 28), and the third treated with the same regimen intravenously. RESULTS: All bladder tumors of the rabbits in the IA group decreased in size, and 3 of the tumors totally disappeared (37.5%). There was also no evidence of lung metastasis. All tumors in the rabbits in the i.v. treatment group increased in size (tumor volume of IA vs. i.v., P = 0.008) and 2 rabbits had lung metastases. All tumors of the control group increased in size and all rabbits had lung metastases. The concentrations of platinum and pirarubicin in the bladder tumors were significantly higher in the IA treatment group than those in the i.v. treatment group at time points from 5 to 10 minutes (P < 0.05) after drug infusion. CONCLUSION: The antitumor effect of IA may be due to higher drug concentrations in the early stage after drug delivery, and the initial circulation of high concentrations of drugs may be the most important factor in suppressing tumor growth. PMID- 9354954 TI - Cessation of spermatogenesis in juvenile spermatogonial depletion (jsd/jsd) mice. AB - BACKGROUND: Mice homozygous for the jsd (juvenile spermatogonial depletion) allele are sterile because they become azoospermic. The onset of such azoospermia was investigated by histologic analysis of sections of testes from jsd/jsd mice. METHOD: The testes removed from C57BL/6-jsd/jsd mice aged 3 to 10 weeks were examined microscopically. RESULTS: At 3 weeks of age, spermatocytes were seen in most of the seminiferous tubules of jsd/jsd mice. However, the number of tubules that contained spermatids was significantly smaller than that counted in the wild type mice. Since degenerative figures were not abundant in the jsd/jsd testes, the decreased number of spermatids found in the tubules suggested a longer duration of development from spermatocyte to spermatid in jsd/jsd mice. The abnormality extended to the development of type B spermatogonia, and a decrease in their number became apparent after 6 weeks of age in most of the jsd/jsd tubules. However, as early as 3 weeks of age, a few seminiferous tubules in jsd/jsd mice already contained only Sertoli cells and type A spermatogonia. CONCLUSION: It is assumed that the decrease in type B spermatogonia occurred at various ages and locations. The defect of spermatogenesis in jsd/jsd mice was attributable to aberrations in multiple steps of spermatogenesis. PMID- 9354955 TI - Renal fossa recurrence of a renal cell carcinoma 13 years after nephrectomy: a case report. AB - Between 1980 and 1995, we performed a nephrectomy with curative intent on 183 patients with renal cell carcinoma at Nagoya University Hospital. Among these patients, 5 (2.7%) developed renal fossa recurrence (median follow-up, 65 months). We report a case of such a recurrence found 13 years after a nephrectomy for renal cell carcinoma (stage pT3a, pN0, M0). A 62-year-old female presented with a nodule on her back. Computed tomography and magnetic resonance imaging revealed a mass in the right back and retroperitoneum, and a biopsy revealed the tumor to be a renal cell carcinoma. Complete resection was performed, followed by administration of alpha-interferon. The patient is doing well 16 months after the operation. The case illustrates that very long-term follow-up after a nephrectomy is mandatory for patients with perinephric invasion of a renal cell carcinoma due to the risk of renal fossa recurrence. PMID- 9354956 TI - Distal ureteral atresia associated with crossed renal ectopia with fusion: recovery of renal function after release of a 10-year ureteral obstruction. AB - We report on a 10-year-old boy with distal ureteral atresia associated with crossed renal ectopia with fusion. He was admitted with a high fever associated with a urinary tract infection. The diagnosis was established by antegrade and retrograde pyelography. The upper hydronephrotic portion of the kidney, obstructed for 10 years, recovered its function after nephrostomy placement. To our knowledge, this is the first patient whose renal function has recovered despite an ureteral obstruction of 10-years' duration. Therefore, we recommend a transient nephrostomy placement even for far advanced pediatric hydronephrosis, to test for the possibility of functional recovery. PMID- 9354957 TI - Extravesical tumor implantation caused by perforation during transurethral resection of a bladder tumor: a case report. AB - We report a case of invasive bladder cancer in which cancer dissemination occurred through a perforation of the vesical wall during transurethral resection of the tumor. A radical cystectomy was performed 1 month later and several clusters of viable cancer cells were histologically identified in a fibrous foreign body granuloma in the paravesicular adipose tissue of the lymphadenectomy specimen. The patient received adjuvant chemotherapy, but developed right inguinal lymph node metastasis 21 months after cystectomy. PMID- 9354958 TI - Brain metastasis from prostate cancer: a case report. AB - Brain metastases from prostate cancer are rare in postmortem examinations, and even rarer in clinical series. We report an unusual case of brain metastasis from prostate cancer confirmed by antemortem diagnosis in a 72-year-old man. The metastatic brain tumor was surgically resected and the patient was kept stable for more than 19 months after diagnosis of the brain metastasis. PMID- 9354959 TI - Primary signet ring cell adenocarcinoma of the prostate treated by radical prostatectomy after preoperative androgen deprivation. AB - We report a case of primary signet ring cell adenocarcinoma of the prostate gland in a 76-year-old man. Radical prostatectomy was performed 4 months after bilateral orchiectomy, which proved to be a successful preoperative androgen deprivation therapy. To date 3 years have passed with no clinical or serologic evidence of recurrent disease. Immunohistochemical testing showed the cancer tissue to be positive for prostate specific antigen and negative for carcinoembryonic antigen. This is the first reported case of primary prostate signet ring cell adenocarcinoma treated by radical prostatectomy, and this mode of therapy should be considered as a treatment of choice for the disease. PMID- 9354960 TI - Hemangioma of the prostatic urethra: hematospermia and massive postejaculation hematuria with clot retention. AB - The case of a 53-year-old man with hematospermia and massive postejaculation hematuria that caused urinary retention is described. This is the sixth case in the English and Japanese language literature. Cystourethroscopic examination revealed that a solitary raised tumor was present just distal to the vermontanum, and that bleeding was from its apex. Histologic examination of an excisional biopsy sample showed features compatible with hemangioma. PMID- 9354961 TI - Glioblastoma multiforme after radiotherapy for metastatic brain tumor of testicular cancer. AB - A patient with left testicular cancer and metastases to retroperitoneal lymph nodes, lung, and brain was treated by chemotherapy, radiotherapy and surgery, and obtained the state of no evidence of disease, but 10 years after radiotherapy, a glioblastoma multiforme tumor appeared in the brain. This is the first report of a glioma appearing after radiotherapy in a testicular cancer patient. PMID- 9354962 TI - Testicular microlithiasis associated with teratocarcinoma and intratubular germ cell neoplasia: a case report. AB - Testicular microlithiasis is a rare condition that has characteristic sonographic and histologic features. It is often associated with premalignant changes and malignant neoplasms of the testes. We report a case of testicular microlithiasis associated with teratocarcinoma and intratubular germ cell neoplasia. Three previously reported cases documented development of malignant germ cell tumor during the clinical follow-up period of patients with testicular microlithiasis. We think that testicular microlithiasis is strongly associated with testicular neoplasms, and that coexistence of intratubular germ cell neoplasia and malignant germ cell tumors with testicular microlithiasis is common. PMID- 9354963 TI - Sonographically-detected impalpable testicular cancer with retroperitoneal bulky metastases: a case report. AB - We report a case of a sonographically-detected impalpable embryonal cell carcinoma with bulky retroperitoneal metastases. A 34-year-old man, who presented with left flank pain, was presumed to have an extragonadal retroperitoneal germ cell tumor. Scrotal sonography revealed a hypoechoic lesion, 7 mm in diameter, which was histologically diagnosed as a primary embryonal cell carcinoma. Evidence suggested that the primary tumor had grown slowly, as the tumor was well encapsulated. This case suggests that some extragonadal germ cell tumors arise from a primary testicular cancers, and that successful treatment of these tumors should include consideration that they may have arisen as a primary testicular mass. PMID- 9354964 TI - Detection of antibodies against cell wall-associated antigens of Mycobacterium tuberculosis (SIHV) by an enzyme linked immunosorbent assay. AB - We have used the cell wall-associated proteins of Mycobacterium tuberculosis (SIHV strain) as antigens in the serodiagnosis of pulmonary tuberculosis. Sera from 19 relapsed and 46 newly diagnosed cases of pulmonary tuberculosis and 21 healthy individuals were tested against the cell wall-associated proteins of M. tuberculosis (SIHV) by an ELISA technique. The results showed 92% and 100% positive titers in new and relapsed cases of tuberculosis, respectively. Control sera analyzed exhibited a negativity of 90%. Cell wall-associated proteins of M. tuberculosis were found to be useful in serodiagnosis of pulmonary tuberculosis in clear distinction from healthy subjects. PMID- 9354965 TI - Effect of cyclosporin A on tissue lipid peroxidation and membrane bound phosphatases in hyperoxaluric rat and the protection by vitamin E pretreatment. AB - The effect of cyclosporin A, a highly effective immunosuppressant, was investigated on hyperoxaluric rats with and without vitamin E pretreatment. Hyperoxaluria was induced by oral feeding of 3% ammonium oxalate in water for 3 days. Cyclosporin A (50 mg/kg body wt.) was administered for 3 days. Pretreatment with vitamin E (50 mg/100 g body wt., once a week for 3 weeks) was carried out before the administration of cyclosporin A and ammonium oxalate. Nonenzymatic ascorbate-induced lipid peroxidation was increased to 1.55-fold in either cyclosporin A-administered or hyperoxaluric rat kidney and liver when compared to control. The lipid peroxidation was further elevated to 1.9-fold when both cyclosporin A and ammonium oxalate were coadministered. The activities of renal and hepatic ATPase, glucose-6-phosphatase as well as the concentrations of thiols were decreased significantly (p < 0.001) when cyclosporin A was administered under hyperoxaluric condition. On pretreatment with vitamin E the cyclosporin A induced biochemical changes observed in the presence of hyperoxaluria were abolished. PMID- 9354966 TI - Antibody responses against Em18 and Em16 serodiagnostic markers in alveolar and cystic echinococcosis patients from northwest China. AB - Western blot analysis was carried out in order to evaluate new serodiagnostic markers, Em18 and Em16, for differentiation of alveolar echinococcosis (AE) from cystic echinococcosis (CE) using 36 serum samples from hydatid patients from Xinjiang, China, where AE and CE are both endemic and one double infection case has been reported. All AE cases except one (5/6) who exhibited a calcified lesion and a single case of double infection showed antibody responses against Em18 and Em16. Some of CE patient sera (6/22) showed antibody response against Em16 except one who showed that against Em18. Analyses of IgG subclass responses against Em18 and Em16 were carried out using all serum samples showing antibody responses against Em18 and/or Em16 (seven CE, five AE, and one AE + CE) and additional samples of three CE and 22 AE from Sichuan, China. IgG4 was the most predominant antibody subclass. Em18 and Em16 were recognized by both IgG4 and IgG1 (in most cases) or by either IgG4 or IgG1 (in minor cases) or by IgG3 (in very rare cases). Neither Em18 nor Em16 was recognized by IgG2 antibodies. The usefulness of Em18 and Em16 as potential new markers for serological differentiation of human AE and CE, respectively, is discussed. PMID- 9354967 TI - Detection of influenza virus RNA in peripheral blood mononuclear cells of influenza patients. AB - Gastroenteritis, arthralgaia and myalgia are frequently associated with influenza virus infections in humans. One explantation for these symptoms may be extrarespiratory transmission of virus by peripheral blood mononuclear cells (PBMC). We tried to detect genomic viral RNA of the nucleoprotein (NP) and H3 subtype hemagglutinin (HA) genes by the method of RT-PCR in PBMC of 18 children aged 1-14 who suffered from an influenza outbreak in the Kansai district of Japan between December 1992 and February 1993. Three of the 18 samples were RT-PCR positive. The NP gene sequence observed in one patient's PBMC was identical to that obtained from his throat swab fluid. The HA gene sequences observed in the two other PBMC differed from those of RT-PCR-amplified DNA from throat swabs by an order of 3-9 nucleotides. We believe these results suggest the presence of a PBMC-associated virus. PMID- 9354968 TI - IFN-gamma-mediated protection against intracerebral challenge with Bordetella pertussis in mice. AB - Mice inoculated with whole cell pertussis vaccine (WCV) acquired protection against intracerebral challenge with Bordetella pertussis without any appreciable antibody production against pertussis toxin or filamentous hemagglutinin. Spleen cells from mice immunized with WCV produced a significant amount of IFN-gamma upon stimulation in vitro with WCV. Furthermore, mice inoculated with recombinant IFN-gamma along with a suboptimal dose of WCV survived longer than those that received WCV alone. These results suggest that, in WCV-immune mice, IFN-gamma plays an important role in protection against intracerebral challenge with B. pertussis. PMID- 9354969 TI - United Kingdom Universities' Research Assessment Exercise 1996: critique, comment and concern. PMID- 9354970 TI - European studies in radiotherapy for breast cancer. PMID- 9354971 TI - Births and deaths in 1996: England and Wales. PMID- 9354974 TI - Compathy: the contagion of physical distress. AB - A qualitative study examining the nurse-patient relationship has identified the contagion of physical distress or 'compathy' as a significant but otherwise neglected phenomenon. Compathy occurs when one person observes another person suffering a disease or injury and experiences in one's physical body a similar or related distress. Thus, compathy is the physical equivalent to empathy. Although the contagion of physiological (compathetic) responses has been previously documented (for example, as couvade or psychogenic epidemics of the workplace), it has not been theoretically explicated. Triggers for the compathetic response are identified and include observing the suffering, hearing, or reading--or even thinking about--descriptions of the symptoms. The relevance of the compathetic response in the caregiving relationship and the necessity of suppressing the response when inflicting pain as a part of providing therapy are described. The concept of compathy is analysed and defended. PMID- 9354975 TI - Uncertainty in mothers' care for their ill children. AB - The purpose of this paper is to present the experience of mothers as they cared for their young children during illness episodes, prior to a diagnosis of asthma in the children. A grounded theory study was conducted with 12 families with asthmatic children, involving three rounds of interviews. In these interviews the mothers revealed that the prediagnosis phase of their children's illness was an overwhelming time for them. Groping in the dark is the core variable that reflects (a) the harrowing experiences of sleepless nights while mothers watched their children struggle to breathe, (b) the wearing toll incurred while helping their children through seemingly unending illnesses, and (c) their increasingly intensified search for answers to resolve their children's health crisis. PMID- 9354976 TI - Combining the analyses of three qualitative data sets in studying young caregivers. AB - Adolescent care of the adult with cancer led the authors to choose three qualitative methods to describe this unexplored phenomenon. In this study, phenomenology, ethnography and unstructured survey were combined to provide a more complete picture of the phenomenon. Data from interviews with 11 youngsters within seven family units, observations, and unstructured questionnaire, demographic data from and field notes were analysed and combined. The processes used in designing and conducting the study and analysing the data, rather than the findings, are emphasized. The data obtained by using these three methods have laid the foundation for further nursing research on caregiving by youngsters and raise questions about combining analyses of three qualitative data sets. PMID- 9354977 TI - Self-coherence, emotional arousal and perceived health of adult children caring for a brain-impaired parent. AB - An important concern for nurses is the ability of adult children to provide effective care to a dependent parent without sacrificing their own health and well-being. The purpose of the study was to examine 'sense of self-coherence' as an inner resource for the attenuation of distress in a sample of 168 adult children who were involved with the care of a brain-impaired parent. Subjects were interviewed twice in their homes in order to obtain data on variables for: self-coherence, emotional arousal, perceived health, and crisis. Findings from the study indicate that adult children with crisis experience in the previous 6 months of caregiving had higher scores for emotional arousal, lower scores for self-coherence, and lower ratings of perceived health than did adult children with no crisis experience. In addition, there was a negative relationship between self-coherence and emotional arousal and a positive relationship between self coherence and perceived health. Both of these relationships were significantly stronger in the presence of crisis experience than in the absence of crisis experience. Finally, there was a negative relationship between emotional arousal and perceived health that was equally apparent in both the presence and absence of crisis experience. The findings suggest that self-coherence is an inner resource that emerges in filial crisis to modulate the emotional impact of the situation. The implication is that measures of self-coherence could be used to assess an adult child's preparedness to appraise and cope with emotional responses to filial crisis events. This information could help nurses anticipate and target resources for vulnerable adult children so that they are less adversely affected by the demands of parent care. PMID- 9354979 TI - Nursing ideology and the 'generic career'. AB - A review of the concept of ideology in a nursing context is presented. The meaning of a nursing ideology is explored and that meaning compared to the proposals for the future of nursing contained in the 1996 report on The Future Healthcare Workforce published by the University of Manchester, England. The paper suggests a number of core beliefs and values that are generally supported in nursing and their implications for the future development of the profession. These include a commitment to the role of science in nursing, the concept of caring in nursing practice and the continued pursuit of the professional project. The paper will demonstrate the extent to which the proposals contained within the report for a 'generic career' compliment or conflict with the suggested ideology of nursing. It concludes with a reference to the policy-making process at local and national levels and the opportunities available to nurses to influence the direction of change. The paper reflects the volatility of the current political climate in health care and the extent to which nurses operating in that climate feel empowered or disenfranchised. PMID- 9354978 TI - Major strain and coping strategies as reported by family members who care for aged demented relatives. AB - This study is based on a previous investigation into 46 caregivers' experience of burden and burnout when caring for a demented elderly relative. The aim of this study was to describe caregivers who develop or experience burnout (group A) and caregivers without experience of burnout (group B) and how they cope with major strain. The interviews focused on the caregivers' descriptions of their major strain (the demented person's memory difficulties and change in behaviour and the caregivers' experiences of their feelings of loss and their new role) and what they did, thought and felt in these situations. The interviews were coded and categorized and a chi-square test was performed. It was found that those in group A more often used an emotion-focused strategy (grieving, worrying and self accusation). They were also the only ones using wishful thinking and stoicism as strategies. Those in group B used a problem-focused strategy more often (confronting the problem, seeking information and seeking social support). Another interesting finding was that the caregivers in group B frequently used the emotion-focused strategy of acceptance in combination with seeking information and seeking social support. To mix approaches like these seems to be an effective choice of strategy. It was also found that the caregivers' gender seems to have an effect on the coping strategy. The demented person's domicile did not, however, appear to result in any significant difference. PMID- 9354980 TI - Identifying the health needs of elderly people using the Omaha Classification Scheme. AB - An examination of what constitutes assessment would help to clarify those aspects which are essential for establishing a suitable nursing plan for clients. There is no consensus on one assessment tool in nursing, and many methods are used. The important areas in a comprehensive assessment (physiological, mental, social and environmental factors) which need to be addressed are covered by the Omaha Classification Scheme (OCS). This paper discusses the application of a research tool (OCS) for assessing the health needs of a given population. The primary focus of this paper is upon the methodology used in the study. A random sample of 500 elderly people was drawn from all elderly people aged 65 years and over, who lived in the rural areas of Isfahan province, Iran. Data were collected to identify the health needs of the population under study using the OCS. The reliability of this research tool was tested for quantitative scores using Cronbach's alpha test. The results showed an acceptable score for reliability (r = 0.78). PMID- 9354981 TI - Therapeutic activities for people with dementia--what, why ... and why not? AB - This paper reviews the literature pertaining to the provision of therapeutic activities for elderly people suffering from dementia. It relates both the provision of such activities, and the types of activity done, to the values and beliefs held by nurses and other professional carers. The range of therapeutic activities cited in the literature is explored, and reasons why activities are sometimes not offered to patients are discussed. Finally, research findings into the effectiveness and value of therapeutic activities for this group of people are reviewed. PMID- 9354982 TI - Elderly patients' perceptions of their spiritual needs and care: a pilot study. AB - This paper describes a pilot study, in a small sample of elderly patients, designed to ascertain their perceptions of their spiritual needs and care. According to the nursing literature, spiritual care is part of the nurse's role. But it is not clear what spiritual needs are or how nurses are expected to give spiritual care. Ten patients from a care of the elderly assessment unit located in a hospital in Edinburgh, Scotland were interviewed about their spiritual needs in the summer of 1995. Eight patients admitted to having experienced spiritual needs at some time in their lives, six while in hospital. The types of needs experienced related to religion, meaning, love and belonging, morality, and death and dying. Their spiritual needs could have been better met if, for example, a quiet room for reflection/prayer had been available and if they had been told about hospital church services and provided with transport to attend. Although limited, the findings contribute to our understanding of spiritual need and spiritual care from the elderly in-patients' view point. Further research, however, is needed to explore the type of spiritual help other elderly patients and other patient groups in different geographical locations feel they would like. PMID- 9354983 TI - Telephone hotline assessment and counselling of suicidal military service veterans in the USA. AB - Studies show that suicide occurs more frequently among people who are elderly, male, single, divorced or widowed, alienated, and among those with a life threatening illness. Military service veterans are not spared these conditions; in some respect, they represent the 'down and out', the lonely and, increasingly, the older isolated people. This correlational descriptive study sought to identify the characteristic profile of telephone hotline users among veterans, their triggering crisis events, and whether the methods commonly used in suicide attempts relate to certain types of crisis. The random sample consisted of 271 veterans of the US military service, ranging in age from 20 to 79 years. Data were collected from nursing notes documented in the hotline suicide telephone call assessment records. The findings portray a sociodemographic profile of military veterans at risk of suicide attempts. Loneliness, alcoholism and unemployment topped the list of triggering events. The most common method used was drug overdose; shooting was a close second. These findings could serve as a base for development of suicide-prevention-focused programmes and optimal use of telephone hotlines for assessment and timely intervention of persons in great crisis. PMID- 9354984 TI - Evaluation of a nurse-led continence service in the south-west of Glasgow, Scotland. AB - This paper reports the results of a pilot study of a nurse-led continence promotion service in both the community and a local nursing home. Telephone and written referrals were made to the service from 28 primary care teams in Glasgow, Scotland. In the nursing home all patients were assessed and an appropriate management plan implemented. A full assessment was carried out in all community patients, including an appraisal of contributory factors, urinalysis and diaries of food and drink intake. A management plan suited to the patient was then implemented. Patients' levels of incontinence in both arms of the study were assessed objectively using the Lagro-Janssen method. The cost incurred in both arms of the study were measured. There was a 69% improvement in the level of incontinence in the community group compared with 30% in the residents wing and 13% in the hospital wing. The savings in the nursing home amounted to Pounds 4152 in the residents' wing and Pounds 1959 in the hospital wing. In summary, a nurse dedicated to urinary incontinence in the community allows improved management, a greater level of awareness and results in resource savings, whilst increasing patient accessibility to a service. PMID- 9354985 TI - Empowering practice nurses in the follow-up of patients with established heart disease: lessons from patients' experiences. SHIP Collaborative Group. Southampton Heart Integrated care Project. AB - This paper reports on the views of patients with established heart disease of a structured programme of follow-up care provided by practice nurses (PNs) in general practice in England. It is based on in-depth interviews with 22 patients receiving an integrated primary and secondary care intervention being developed and piloted for patients following heart attack or diagnosis of angina. Patients identified the important features of follow-up care to be easy access to a health professional who possessed knowledge and social and emotional skills. A range of views about the ability of PNs to provide such care emerged from patients' accounts. Patients' perceptions about the seriousness of their condition and the way PN follow-up care was provided in practices emerged as important issues affecting patients' views. In addition, perceptions about the practice nurse's role, status and knowledge, existing relationships with general practitioners, and issues of communication were also important factors. It is concluded that in order to develop high-quality PN-led services for patients with established heart disease, four issues need to be taken into account: practice nurse training; continuity of follow-up care; the integration of the primary and secondary care interface; and development of the practice nurse's status within the primary health care team. PMID- 9354986 TI - Disability and oppression: some implications for nurses and nursing. AB - Since the 1970s organizations of disabled people have defined disability as a form of oppression. However, despite nurses (along with other professionals) increasingly being viewed as part of this experience of oppression, British nursing literature has to date largely failed to conceptualize disability in these terms. In this paper the five faces of oppression identified by Young are modified to include discrimination and developed to provide a framework in which evidence of the oppression experienced by disabled people may be analysed. It is argued that if nurses and the nursing profession are to challenge and reduce oppression then this should be via a process of awareness of the nature of disability, reflection on practice and development in partnership. Accordingly, an agenda is identified which has implications for nursing practice, education and research. It is stressed that if nursing is not to be viewed as irrelevant to the needs of disabled people or as part of the process by which they are oppressed then it is vital that this agenda is heeded. PMID- 9354987 TI - A personal reflection on the nature of somatic treatments and the implications for mental health nursing. AB - Somatic treatments in psychiatry, ECT (electro-convulsive therapy) and pharmacotherapy, are based on a particular conception of our nature as human beings, and on a particular conception of clinically constructed reality. Acceptance of the basis for such treatments constitutes a tacit admission of our nature as being essentially materialist and deterministic biological machines, and undermines the connectedness which ensures that we each acknowledge the humanity and humanness of the other. By objectifying the other as different (by reason of disordered biology), these connections are severed and allow the imposition of physical forms of treatment in the name of cure. In this personal reflection, I draw upon my own experience of ECT from a nursing perspective, and attempt to explain the basis of my antipathy for this treatment. In the course of this exploration I also raise some of the issues which I feel nursing must deal with if it is to continue uncritically to adopt the biological paradigm as the basis of its praxis. PMID- 9354988 TI - A challenge for community psychiatric nursing: is there a future in primary health care? AB - The growing debate surrounding the role of the community psychiatric nurse (CPN) in the United Kingdom is reviewed. Issues which have attracted significant interest and which form the focus of this paper are the prioritization of CPN services, CPN attachment to primary health care (PHC), and the effectiveness of clinical interventions. The requirement for CPNs is now to concentrate services on people experiencing severe and enduring mental health problems. Innovative and effective clinical and social interventions for this client group are beginning to disseminate into everyday CPN practice. Problem-solving family interventions, cognitive therapies and case management are three such examples. The past, present and possible future role for CPNs working in primary health care settings with people experiencing nonpsychotic mental health problems is a particular focus in this paper. Drawing on the relevant literature, central issues addressed are the process and outcome of CPN work with nonpsychotic service users, reasons for the growth of CPN involvement in PHC, and the overall expansion of interest in mental health interventions within the primary health care environment. The literature suggests that this expansion has been strategically unplanned, but that mental health need amongst primary health care service users is significant. The concluding contention of this paper is that a future role for CPNs in primary care does exist. PMID- 9354989 TI - Assessing quality in health care services: lessons from mental health nursing. AB - Quality in health care services is neither a simple nor an apolitical issue. Quality is a complex concept and how it is defined and measured has important consequences for services. The definition and measurement of service quality in health care has slipped from professional toward managerial control. Professionals such as nurses have an absolute concept of quality which is a part of their value system and based upon their training and experience. Managerial concepts of quality may be influenced by other organizational concerns such as the pursuit of efficiency. This paper outlines quality assurance systems in mental health nursing and the wider quality debate and argues that there may well be a trade-off between quality of service and efficiency. The moves toward outcome measures and performance indicators are also discussed and the argument forwarded that such a focus serves to exclude users and cloaks serious issues in service delivery. The argument of this paper is that evaluation of service quality needs to include users, and this could provide them with a platform for enhanced involvement in service planning. PMID- 9354990 TI - Focus groups: issues of analysis and interpretation. AB - Focus groups have become a popular method in nursing research. Their history can be traced back to marketing research methods, but they have also been used in qualitative, ethnographic research. Our study, which used this approach as part of data collection, raised many issues of analysis and interpretation: in particular, the importance of paying attention to the sequence of focus group discussions, the individuals involved, and the social context of the focus group. We conclude that focus groups are not a 'quick and easy' method of collecting data, and that issues of validity and the relationship between focus group data and other data require careful consideration. PMID- 9354991 TI - Different approximations of the McGill Pain Questionnaire in the Norwegian language: a discussion of content validity. AB - The McGill Pain Questionnaire (MPQ) is a well recognized measuring instrument for pain in English-speaking countries. Several efforts have been made to develop equivalent pain-measuring instruments in other languages. However, the method of translating the English words contained in the MPQ into another language implies that questions about validity may be posed. In Norway three different pain questionnaires have been developed which are inspired by the MPQ. A primary focus for developing the Norwegian Pain Questionnaire (NPQ) was the semantics of pain; the focus of the adapted MPQ was to include commonly used somatosensory Norwegian descriptors of pain in the population of patients with low back pain; the Norwegian McGill Pain Questionnaire (NMPQ) was literally translated into Norwegian to provide a equivalent pain questionnaire to the MPQ for cross cultural comparisons of pain. Examination of content validity of the adapted MPQ and the translated version of the MPQ is examined by comparing the words in those questionnaires with words collected among Norwegians in the process of developing the NPQ. The findings support the content validity of the adapted MPQ. The NMPQ, however, should be further refined to better fit the semantics of pain in Norway. PMID- 9354992 TI - An ethnomethodological analysis of the use of seclusion. AB - The use of seclusion in psychiatric practice is a contentious issue. This paper reports on an ethnomethodological approach to understanding the decision-making process in the use of seclusion. Through first-level and second-level reasoning analysis, three themes emerged which underscored the subjects' rationalization process. These were: (a) mechanistic searching, (b) frame conflict, and (c) asylum status. This research suggests that the decision to use seclusion is based on a complex interplay of cultural and organizational factors rather than due to the presentation of symptoms by the patient. PMID- 9354993 TI - Sampling hard to reach populations. AB - Studies on 'hidden populations', such as homeless people, prostitutes and drug addicts, raise a number of specific methodological questions usually absent from research involving known populations and less sensitive subjects. This paper examines the advantages and limitations of nonrandom methods of data collection such as snowball sampling. It reviews the currently available literature on sampling hard to reach populations and highlights the dearth of material currently available on this subject. The paper also assesses the potential for using these methods in nursing research. The sampling methodology used by Faugier (1996) in her study of prostitutes, HIV and drugs is used as a current example within this context. PMID- 9354994 TI - The effects of an HIV/AIDS educational programme on the anxiety level of nursing students. AB - Acquired immunodeficiency syndrome (AIDS) is a serious disease that has special concern for the health care provider. AIDS has continued to grow despite control efforts. As the disease infectivity period remains lengthy, and the heterosexual population is affected to a greater degree, the level of anxiety has also risen despite educational endeavours. Many fears and anxieties have been associated with AIDS patients by health care workers. The reduction of stress, perceived risk and discomfort following educational efforts have been supported in past research. Educational programmes will need to be given for current health care workers at all levels as well as nursing students. Future nurses must be prepared to meet this challenge. This study was conducted using a convenience sample of nursing students at a university in western United States. Its purpose was to assess any changes that occurred in state anxiety following an educational presentation. Spielberger's State-Trait Anxiety Inventory was used as the measurement instrument. Some anxiety levels were significantly reduced. PMID- 9354995 TI - Nursing knowledge, theory and method revisited. AB - With the approach of the 21st century, nursing is having to respond to diverse influences which are remoulding the professional landscape. Not least of these is the changing status of western economies which underpins a drive towards evidence based practice and an increased emphasis on multidisciplinary approaches to health care delivery. Certainty in health care is now a thing of the past. Central to the way the nursing profession embraces the future is its underlying philosophy: that which articulates professional values and shapes practice, research, education and management. In a time of change it is therefore essential to revisit the philosophical framework which underpins nursing. The debate in nursing research and theory appears to have stressed the polarization of viewpoints. It may be the case that feminist writers, ethnographers, positivist researchers and nursing theorists, in defending their own points of view, diminish rather than enhance professional dialogue. This paper reviews the nature of this debate within nursing and considers the implications that a dichotomous position may have for knowledge, theory and research method within the current context of health care. It then suggests a philosophical framework which could be relevant and accessible across the whole spectrum of nursing activity. In so doing, the paper aims to contribute to the discussion around epistemology and method in a way which encompasses the diversity found within the broad church of nursing. PMID- 9354996 TI - Assessing and preparing students for distance preceptorship placements. AB - Preparing students for distant preceptorship placements requires knowledge of individual students through formal and informal mechanisms. Since they receive support via telephone, fax or mail, the first onus is on students to manage their preceptorship situations effectively. In order to enhance their abilities in preceptorship situations, students can be assessed for their learning and personality styles. Findings from two style assessment tools, Kolb's Learning Style Inventory and Myers-Briggs Type Indicator, can be used to gain insight into students' interactions with preceptors and to assist instructors in giving the right kind of support and direction after students are placed. PMID- 9354997 TI - Nursing students' and faculty's perceptions of the characteristics of 'best' and 'worst' clinical teachers: a replication study. AB - This replication study described and compared the characteristics of 'best' and 'worst' clinical teachers as perceived by 185 Greek students of nursing and 31 clinical teachers. It is a replication of Mogan & Knox's (1987) study, which has also been replicated by Nehring (1990). The Nursing Clinical Teacher Effectiveness Inventory (NCTEI) was used for data collection. This tool is a 48 item seven-point-scale checklist which describes discrete teacher characteristics grouped in five categories or subscales. Subjects were invited to rate their 'best' clinical teacher using the NCTEI, and then their 'worst' clinical teacher. Faculty's and students' perceptions agreed with most of the highest-rated characteristics of the 'best' clinical teachers. There was less agreement in the lowest-rated characteristics of the 'worst' teachers. There were not significant differences between the rating of students and faculty when categories of characteristics were compared, with the exception of the category 'interpersonal relationship' of the 'worst' teachers. The most distinguishing characteristics between 'best' and 'worst' clinical teachers for students and faculty in this study and in both Mogan & Knox's and Nehring's samples, were being a good role model and encouraging a climate of mutual respect. PMID- 9354998 TI - A longitudinal framework for fostering critical thinking and diagnostic reasoning. AB - Nurses' reasoning skills are of interest to nurse researchers, managers, clinicians, educators and students. Recent emphasis on critical thinking has raised important questions regarding the modes of reasoning necessary for practice and appropriate measures to improve nurses' cognitive skills. This paper analyses critical thinking and diagnostic reasoning. A longitudinal framework for promoting both modes of reasoning, beginning with nursing students through to experienced clinicians, is proposed as a model worth testing in nurse samples. PMID- 9354999 TI - Use of the QSR.NUD.IST computer program to identify how clinical midwife mentors view their work. AB - This paper looks at the use of a computer program (QSR.NUD.IST) to analyse qualitative data. The data are derived from interviews with clinical midwife mentors as part of an evaluation of a midwifery educational programme. An account is given of the content analysis of the data with reference to the literature, and then the way the programme was used to analyse the data a second time. A description is given of the way this programme in particular functions using the textual data as an example. The use of computers in general for analysing qualitative data is discussed in terms of their advantages and disadvantages. The results of the study are described briefly showing how the mentors view their role and responsibilities. PMID- 9355000 TI - Isolated revision acetabuloplasty using a porous-coated cementless acetabular component without removal of a well-fixed femoral component. A 3- to 9-year follow-up study. AB - The results of isolated acetabular revision performed in 31 patients (32 hips) were monitored for between 3 and 9 years. All femoral components were well fixed and not removed or revised at the time of index surgery. There were 4 hips with little or no acetabular bony defect, 2 hips with pure segmental defects (type I), 10 hips with cavitary defects (type II), 15 with combined segmental cavitary defects (type III), and I with pelvic discontinuity (type IV). All revision acetabular implants were cementless, using a porous-coated hemispheric cup with or without bone-graft. There were four grade I reconstructions, 16 grade II reconstructions, and 12 grade III reconstructions. At final follow-up evaluation 94% of the cups were judged to be stable. Two hips required a second revision acetabuloplasty because of loss of fixation of the cup. The 2 repeat revisions were also done without removal of the femoral component. One acetabular component had evidence of rotational migration, which stabilized and remained nonprogressive. There were no cases of femoral component radiographic or clinical failure. The mean pre and postoperative hip scores were 44 and 83, respectively. The pre- and postoperative pain scores were 12 and 42, respectively. The findings of this study suggest that isolated acetabular revision, using a cementless porous-coated hemispheric cup, can be successfully performed without removing or revising a well-fixed femoral stem and not compromise the final outcome. PMID- 9355001 TI - The Stanmore total hip arthroplasty. A 15- to 20-year follow-up study. AB - An ongoing study was made of 804 primary Stanmore total hip prostheses implanted in 839 patients between 1973 and 1991. The earliest surviving implants were brought back for radiologic and clinical review in 1995 at an average of 17 years after surgery. The remainder of the patients still living were sent a questionnaire to assess their current status. Survivorship was 95% at 10 years, 85% at 15 years, and 73% at 20 years. The average Merle d'Aubigne-Postel score was excellent up until 14 years. Patient satisfaction remained high until 22 years. Overall, 10% of the prostheses had failed. The results of this study suggest that the Stanmore prosthesis is capable of producing satisfactory long term results that compare favorably with those of other cemented prostheses. PMID- 9355002 TI - Resurfacing of only the femoral head for osteonecrosis. Long-term follow-up study. AB - Fourteen patients (21 hips) with osteonecrosis of the femoral head with collapse had the femoral head resurfaced with a cemented titanium shell. All of the femoral heads were Ficat stage III or IV. Of the 21 surgeries, 7 were failures. Treatment for all 4 patients with sickle cell disease or trait failed (100%). When the cases of 17 patients who did not have sickle cell disease or trait were reviewed separately, the success rate was 14 of 17 (82%). The follow-up periods (all > 5 years) of the 14 successful patients in this group averaged 6.2 years, and their average Harris hip score was 87 (10 excellent, 4 good). Of the 14 successes, 10 patients had a follow-up period longer than 5 years (average, 7.7 years) and an average Harris hip score of 94 (7 excellent, and 3 good). There was no evidence of loosening and there was no osteolysis. It is concluded that this operation provides an alternative to hemiarthroplasty, total joint arthroplasty surgery, or bipolar arthroplasty. This is a time-buying first-stage operation and, for younger patients, will not last a lifetime. The concept appears prudent because the surgical procedure is directed at the site of primary disease, the femoral head. PMID- 9355003 TI - Loosening of massive proximal femoral cemented endoprostheses. Radiographic evidence of loosening mechanism. AB - A review of the radiographs of 24 patients with massive proximal femoral cemented tumor endoprostheses revealed a repetitive radiographic sequence that culminated in implant loosening. The initial step was osteolysis of the proximal femur at the bone-prosthesis junction (13 of 24 cases, 54%). The osteolysis then progressed into the adjacent periprosthetic bone-cement interface as a radiolucent line (5 of 24 cases, 21%), followed by extension along the stem, culminating in implant loosening (4 of 24 cases, 17%). Two prostheses (8%) have required revision for loosening. The proximal osteolysis was noted prior to the appearance of progressive periprosthetic radiolucent lines in 11 patients and simultaneously in 2 but never followed periprosthetic radiolucent lines (P < .05). These findings are consistent with loosening by debris-induced osteolysis. This loosening sequence, confirming previous observations of cemented component loosening, should encourage efforts to prevent or delay the onset of this sequence. PMID- 9355004 TI - The cement mantle in the Exeter impaction allografting technique. A cause for concern. AB - The postoperative radiographs of 35 patients who underwent impaction allografting of the proximal femur were reviewed. Of Gruen zones that could be clearly visualized, 39.9% contained areas where cement was absent. Even when an adequate mantle was present, cement voids were commonly seen. These cement mantle deficiencies were confirmed in a series of cadaveric impaction allografting procedures. They appear to be a consequence, at least in part, of an inadequate differential between trial and actual component sizes. Additionally, 4 patients were identified with significant component migration secondary to radiographically visible cement mantle fractures within the first 6 months of surgery. It is concluded that the surgical technique requires modification to ensure a more consistent cement mantle and clinical result. PMID- 9355005 TI - An analysis of Food and Drug Administration medical device reports relating to total joint components. AB - A total of 1,717 total hip and 2,769 total knee medical device reports submitted to the U.S. Food and Drug Administration (FDA) from 1984 through 1993 were reviewed. A large percentage of total hip complications could be attributed to some aspect of component modularity. Cementless modular acetabular components were the single largest source of device-related complications. Fifty-six percent of total knee medical device reports (MDRs) were associated with accelerated polyethylene wear. By location, MDRs identified patellar (46%), tibial (33%), and femoral (5%) component complications. It was estimated that less than 5% of device-related complications were reported to the FDA. Based on the reports received, it was apparent that mechanical failure of components was a common and increasing cause of total joint revision. PMID- 9355006 TI - Comparison of cerclage techniques using a hose clamp versus monofilament cerclage wire or cable. AB - To assess the compressive force generated by different techniques of cerclage wiring, the compression produced by 6 different cerclage techniques, including monofilament wires, Dall-Miles cables (Howmedica, Rutherford, NJ), and hose clamps, was measured. Tests were performed on both a polysulfone cylinder model and a cadaver bone model. The hose clamp generated significantly higher compression force than any other cerclage device tested. Doubled wires produced similar compressive force as the cables. As the hose clamp is the least expensive and produces the maximum compression, it may be advantageous in instances in which maximum compression is desirable as a temporary device, but because it is not implantable, it must be replaced by other techniques prior to wound closure. The Dall-Miles cable is expensive and its compressive force can be equaled at lower cost by a doubled wire. The use of a single strand of monofilament wire was the least effective. PMID- 9355007 TI - Kinematics of posterior cruciate ligament-retaining and -sacrificing mobile bearing total knee arthroplasties. An in vitro comparison of the New Jersey LCS meniscal bearing and rotating platform prostheses. AB - The posterior cruciate ligament (PCL)-retaining, meniscal bearing and the PCL sacrificing rotating platform designs of the LCS prosthesis (DePuy, Warsaw, IN) were designed to minimally constrain knee kinematics while minimizing bone-cement prosthesis interface stresses and polyethylene wear. The kinematics and stability of the knee following arthroplasty with these devices rely on adequate tensioning of the remaining soft tissues by management of the flexion/extension gaps at the time of surgery. In this in vitro study, the knee kinematics of the function of the quadriceps mechanism for 8 cadaveric knees were measured quantitatively before and after implantation of these 2 prosthesis designs. Following implantation of the PCL-retaining, meniscal bearing prosthesis, anterior translations during anterior drawer testing were significantly greater (P < .05) than those seen in the intact knee. Implantation of the PCL-retaining, meniscal bearing prosthesis resulted in an increase in the extension gap of 2 mm. Quadriceps force needed to achieve full extension was increased by 30% over that needed in the intact knee. The PCL-sacrificing, rotating platform prosthesis constrained anterior translation such that nearly normal anterior knee stability was reestablished; however, the extension gap was increased by 4 mm and the quadriceps force needed to achieve full extension was 50% greater than that needed in the intact knee. Attempts to achieve joint stability by increases in the thickness of the tibial component to widen the flexion/extension gaps results in compromises of quadriceps efficiency, particularly in the absence of a functioning PCL, as demonstrated in this in vitro model. Patients receiving the PCL-sacrificing prosthesis may experience difficulty in those activities requiring quadriceps power near full extension, such as rising from a chair or ascending or descending stairs. PMID- 9355008 TI - Histologic evidence of cortical allograft bone incorporation in revision hip surgery. AB - There is no published evidence concerning the histology of cortical allograft bone implanted during revision arthroplasty surgery. Five cases are reported in which cortical allograft bone had been used where biopsies were available between 2 and 27 months after implantation. All cases showed favorable histologic features, with soft tissue attachment by fibrous adhesion, union of graft to host, and osseous remodeling. PMID- 9355009 TI - Patellofemoral joint after total knee arthroplasty. Effect on contact area and contact stress. AB - Compressive contact stress between the patella and the anterior femur and between the quadriceps tendon and anterior femur was measured before and after total knee arthroplasty in 5 cadaver knee specimens using a digital electronic sensor. Contact stresses were measured in the normal knee and after total knee arthroplasty with an unresurfaced patella, a dome-shaped patella, and a conforming patella. Patellofemoral contact stresses did not change significantly after total knee arthroplasty when the patella was not resurfaced, but they increased significantly after the patella was resurfaced with both the dome shaped and the conforming components. The conforming patella had the highest contact stresses because it tilted at flexion angles greater than 90 degrees and applied load to a small area on the superior portion of the patellar component. The conforming patella markedly decreased tendofemoral contact force because the thicker superior pole of the patella tented the quadriceps tendon at flexion angles greater than 120 degrees. This further increased patellofemoral contact force in deep knee flexion. PMID- 9355010 TI - Stiffness of trabecular bone of the tibial plateau in patients with rheumatoid arthritis of the knee. AB - Stiffness of subchondral proximal tibial trabecular bone is a factor in the stability of prostheses implanted into that bone. The stiffness of trabecular bone in osteoarthritis (OA) has been documented. Trabecular bone in rheumatoid arthritis (RA) is osteopenic in numerous sites and morphologically abnormal in the proximal tibia. Reliable data on proximal tibial bone in RA are lacking, although 1 study failed to identify abnormalities. The purposes of this study were (1) to document the stiffness of the proximal tibial cancellous bone in patients with RA, (2) to determine the effect of angular deformity on bone stiffness in rheumatoid patients, and (3) to compare RA stiffness values with those in published reports for OA. Fifteen tibial plateau were obtained from patients with RA during surgery. Each plateau was horizontally seated in a mold and covered with cement. The plateau was divided into 6 regions, which were used to facilitate comparison between specimens and the existing literature. Indentation tests were conducted with a 4-mm-diameter cylindrical indentor controlled by an MTS machine. The indentor descended at a rate of 2 mm/min to a maximum depth of 1.0 mm; load and displacement data were digitally recorded. Stiffness was calculated from the slope of the linear region of the curve using best-fit linear regression. Where varus deformity was present, stiffness in the medial plateau was higher overall than for the other compartment; whereas in the case of valgus deformity, stiffness of the lateral side was significantly higher (P < .05 for each observation). In comparison to older normal specimens, both the medial compartment of the varus RA specimens (P < .01) and the posterolateral compartment of the valgus RA specimens (P < .01) had significantly lower stiffness. Comparison with OA specimens showed that in varus RA, the posteromedial region had significantly lower stiffness than in varus OA at the same site (P < .01). In valgus RA, the lateral region had significantly lower stiffness than in valgus OA at the same site (P < .01). The mean stiffness ratio of the valgus RA was significantly (P < .01) altered from normal, and for the varus RA, it was significantly (P < .01) different from normal posteriorly. The stiffness ratios for the varus RA were significantly (P < .01) different from those for varus OA; there was no difference between valgus RA and valgus OA. It is concluded that RA affected bone has significantly lower stiffness than normal and osteoarthritic bone. The loaded plateau is stiffer than the unloaded plateau in angular deformity, but is still less stiff than normal bone and osteoarthritic plateaus with corresponding deformities. PMID- 9355011 TI - Anatomic variation of structural properties of periacetabular bone as a function of age. A quantitative computed tomography study. AB - The shape of the acetabulum, the volume of the periacetabular bone, and its density for 125 patients with a wide age range have been quantified using quantitative computed tomography. The goals were to study the relationship between geometric and densitometric properties and provide normative data for finite-element analysis. Significant correlations were found between acetabular diameter and (1) depth, (2) cancellous periacetabular bone density, and (3) periacetabular total bone volume. Only changes in densitometric properties significantly correlated with age. Sphericity of the acetabulum did not increase with age. Variability in bone morphology and density was found for both male and female groups. Surgeons using purely geometric measures to quantify the integrity of acetabular bone should be aware of their limitations when selecting hardware for total hip arthroplasty. PMID- 9355012 TI - Tumors around implants. AB - In 1989, A. G. Apley recommended cautious surveillance of malignant tumors that developed in association with orthopaedic implants. This retrospective review of the Bristol Bone Tumour Register between 1980 and 1992 reports on 240 malignant soft tissue sarcomas. Eighteen developed in the thigh region of patients more than 50 years old, and 4 of these developed in the soft tissues around a hip arthroplasty. PMID- 9355013 TI - Progressive bilateral pelvic osteolysis in a patient with McKee-Farrar metal metal total hip prostheses. AB - As accumulating evidence indicates that polyethylene plays a central role in periprosthetic osteolysis, there is a renewed interest in alternatives such as metal-metal bearings. Several long-term studies report encouraging results with the McKee-Farrar total hip arthroplasty, but there is a paucity of data on the incidence, severity, and pathogenesis of osteolysis in metal-metal bearing total hip arthroplasties. This study presents a patient who had progressive bilateral pelvic osteolysis associated with his McKee-Farrar metal-metal total hip prostheses. His left hip was revised after 13.5 years of service. The tissues revealed no gross metal staining and fewer inflammatory constituents than are typically found in metal-polyethylene bearing hips. His right hip was still functioning after 22.5 years of service, although the acetabular component was loose by that point. An arthrogram of this hip demonstrated communication of the joint with the iliac osteolysis. The development of osteolysis in both hips followed a pattern similar to that seen in metal-polyethylene total hip arthroplasties. Bearing wear could not be detected in either of the hips. Accumulating evidence indicates that particulate debris of appropriate size and number is capable of fueling periprosthetic inflammation. Specific to this study, consideration should be given to particles of cobalt-chromium alloy, polymethyl methacrylate bone-cement, and barium sulfate. Other factors that should be considered are increased joint fluid pressure, soluble inflammatory mediators, and the effective joint space. When bone becomes part of the effective joint space, it is exposed to particulate debris, soluble factors, and potentially increased joint fluid pressures, which may promote localized bone resorption. It must be kept in mind that the development of osteolysis is multifactorial. Although bearings with better wear characteristics are desirable, the elimination of polyethylene will not eliminate osteolysis. PMID- 9355014 TI - Candida infection after total knee arthroplasty. Management without resection or amphotericin B. AB - A rare case of Candida infection after revision total knee arthroplasty that was treated medically, without amphotericin B or resection arthroplasty, is reported. This case occurred in an elderly patient without predisposing medical problems. The patient was treated with only a suppressive dose of ketoconazole. The patient was last evaluated 6 years after the revision surgery and had no problem or signs of infection. Factors contributing to successful medical treatment in this case were likely the routine debridement at revision surgery and the patient's intact immune system. PMID- 9355015 TI - Nonsurgical management of supracondylar fracture above total knee arthroplasty. Still the nineties option. AB - Supracondylar fracture of the femur above a total knee arthroplasty has been reported to occur in 0.3-2.5% of all cases. The case of a patient who sustained such a fracture subsequent to a fall and whose fracture was managed without surgery with a good result is reported. There was no evidence of coexisting loosening, osteolysis, or significant wear, and satisfactory bony reduction was achieved, maintaining correct alignment of the prosthetic components. Healing occurred in 3 months, and the patient remains under follow-up evaluation with a painless knee and a range of movement similar to prefracture levels with no evidence of implant loosening. Despite the current enthusiasm for internal fixation of these fractures, a review of the current literature revealed that neither conservative nor operative management has a significant proven advantage, and the treatment of these difficult and uncommon fractures remains challenging. Nonoperative treatment of fractures above well-fixed components can, however, be as successful as surgical intervention, and remains a viable first-line approach. Conservative management also lacks the potential risks of operation, while maintaining the option of later surgical intervention if required. PMID- 9355016 TI - Incipient compartment syndrome of the thigh following total knee arthroplasty. AB - Compartment syndrome of the thigh is in itself a rarity because of the large size of the compartment and the relatively high compliance of the thigh which allows accommodation to volume changes due to hematoma or tissue edema. Most cases have been reported in association with impact trauma to the lower extremity or in association with crush syndrome. A previously unrecognized complication of total knee arthroplasty where an incipient compartment syndrome developed in the thigh extensor compartment is reported. PMID- 9355017 TI - Fulminant hepatic failure due to cardiac tamponade associated with mixed connective tissue disease. AB - We describe a 42-year old Japanese woman with mixed connective tissue disease (MCTD) who developed fulminant hepatic failure and hepatic encephalopathy. Massive pericardial effusion accompanying cardiac tamponade was shown by echocardiography. The hepatic failure was considered to be caused by a low cardiac output state because of cardiac tamponade, which might be due to cardiac involvement of MCTD. This is a rare case, showing an unusual progressive course in MCTD. PMID- 9355018 TI - Platelet alpha-adrenoceptor density in patients with left ventricular volume overload. AB - We have evaluated the possibility that, in patients with left ventricular heart disease, the effects of aortic and mitral regurgitation on platelet alpha 2 adrenoceptors may depend on the origin and severity of the overload. Receptor density and binding affinities were estimated by their specific binding to [3H] yohimbine. In blood donor controls (CON), the receptor density was 4.72 +/- 0.41 fmol/10(6) platelets (n = 31). In the volume overloaded patients (n = 35), the total density was elevated by 57% (p < 0.05) accompanied by a 69% (p < 0.05) increase in plasma epinephrine. Compared with CON, patients with pure aortic regurgitation (AVR, n = 12) showed a 91% (p < 0.0001), pure mitral regurgitation (MVR, n = 15) 43% (p < 0.05) and mixed mitral and aortic valve regurgitation (MOL, n = 8) 23% increase in receptor density. Furthermore, the elevation of the density in the aortic disease was significantly greater (p < 0.05) than in the mixed overload group. There was a weak positive correlation between the increase in receptor density and the ejection fractions (r = 0.27), suggesting that the former may be dependent on the severity of the volume overload. The results show that in left heart valvular disease, aortic regurgitation leads to a highly significant increase in alpha-adrenoceptor density, while the effects of mitral valve disease exhibit borderline significance. These findings probably point to the differences in the extent of influence of the origin and severity of the two forms of left ventricular volume overload (LVO), as well as the ensuing hemodynamic changes on the cardiac contractile apparatus. PMID- 9355019 TI - Effect of long-term blood pressure control on salt sensitivity. AB - The objective of this study was to reevaluate salt sensitivity (SENS) after a period of antihypertensive treatment (AT). SENS was measured in ten patients, before and after 18 +/- 6 months on AT. The average for all mean blood pressures (MBP) measured during AT was used as an index of blood pressure (BP) control. After at least eight weeks on placebo only, all patients were submitted to an ad libitum diet (ALD), low salt diet (LSD), and high salt diet (HSD) during one week each. SENS was considered as the percent change of the MBP between the seventh day of LSD and HSD. Weight, BP, and daily urinary Na+ and K+ excretion (mean of seven days) on ALD were the same in the first (F) and second (S) evaluation. SENS did not significantly change from the F and S measurement. An inverse correlation was obtained between individual SENS difference and the average mean blood pressure (AMBP) (r = -0.85, p = 0.0018). In conclusion, patients who showed greater decreases in SENS were the ones with the best BP control. PMID- 9355020 TI - Castleman's disease. A case of angiofollicular lymphoid hyperplasia followed from diagnosis (1985) until 1995. AB - A case report is presented of a patient, (R.C.), who suffered from Castleman's syndrome or angiofollicular lymphoid hyperplasia. The first signs of the disease occurred in 1985 in the form of a left axillary adenopathy. The diagnosis was made after a surgical exeresis of the affected lymph nodes through an histologic test which revealed that it was a case of the plasmocyte version of Castleman syndrome. The histochemical tests showed the monoclonal nature (lambda chains) of the plasmocyte population. Since then the patient, who has been followed every six months with blood chemistries and instrumental check-ups, has not suffered any further relapses. PMID- 9355021 TI - Evaluation of esophageal passage in selected connective tissue diseases. AB - The objective of this study was to determine whether, as in with other types of connective tissue diseases, there exists esophageal passage dysfunction as it exists in systemic scleroderma. It was also interesting to establish whether there is any correlation between this kind of dysfunction and the subjective complaints connected with the esophagus as well as the occurrence of Raynaud's phenomenon. Evaluation of the esophageal passage was performed on the basis of scintigraphic examination. The delay of esophageal passage occurs, besides systemic scleroderma, in other types of connective tissue diseases. This examination is a valuable test, thanks to which changes in the esophagus can be discovered, despite the lack of subjective complaints. In addition to systemic scleroderma, there is no close correlation between dysphagia and Raynaud's phenomenon. PMID- 9355022 TI - Inhibitory effects of ubenimex (bestatin) on the invasion of uterine cervical carcinoma cells and their production and activation of gelatinase A. AB - The present study was undertaken to investigate the effects of the aminopeptidase inhibitor ubenimex (bestatin) on the invasive activity of cultured human uterine cervical carcinoma cells. The invasion of squamous cell carcinoma OMC-1 and SKG IIIb cells, and adenocarcinoma OMC-4 and CAC-1 cells into reconstituted basement membrane (Matrigel) was inhibited by the presence of bestatin in a concentration dependent manner. However, bestatin did not have any effect on tumor cell proliferation or migration. Immunoblot analysis of tumor-conditioned medium showed that the treatment of tumor cells with bestatin resulted in the reduction of the 72 kDa gelatinase level (gelatinase A, latent form) in the four cell lines examined, and the reduction of the 68 kDa gelatinase level (gelatinase A, active form) in SKG-IIIb cells. Bestatin inhibited hydrolyzing activities towards substrates of aminopeptidases in tumor cells, but did not directly inhibit gelatinase A. These results suggest that bestatin may inhibit the invasion of uterine cervical carcinoma cells possibly through the inhibitory mechanisms for production and activation of gelatinase A modulated by tumor aminopeptidases. PMID- 9355023 TI - The value of chest CT scan in the work-up of head and neck cancers. AB - The systemic work-up of patients with head and neck cancers includes a routine chest x-ray and panendoscopy. In patients with a high risk for head and neck cancers, such as heavy smokers and drinkers, a high incidence of second upper aerodigestive tract tumors is expected, in addition to the risk of metastatic spread from their primary neoplasm. In this study, we evaluated the contribution of a chest CAT in patients with negative plain chest x-rays. Twenty-four consecutive patients with head and neck cancers were entered. All patients had locally advanced disease. Nine of the 24 patients (37.5%) had positive findings on their chest CAT scans. All lesions detected by CAT, except for one, were biopsied and reported as squamous cell carcinomas. All these patients had T3 and T4 lesions and only one was N0. Three of these lesions were considered to be metastatic and four more were believed to be second primary lung tumors. In two patients, the nature of the lesions was not determined. In conclusion, because of the high incidence of positive lung CAT scans in this group of patients, we recommend a chest CAT for work-up prior to initiating any definitive therapy for their head and neck cancers. PMID- 9355024 TI - 2-chlorodeoxyadenosine (cladribine) induced allergic cutaneous reactions with eosinophilia in a patient with B-cell chronic lymphocytic leukemia. AB - We present a 68-year old patient with B-cell chronic lymphocytic leukemia (CLL) treated with 2-chlorodeoxyadenosine (2-CdA). This is the first reported patient with B-CLL in whom skin lesions and eosinophilia were observed simultaneously. The most frequent side effect of this drug is myelosuppression with pancytopenia. So far, there have been few reports of cases where either skin reactions or eosinophilia, occurring separately, were observed as side effects from 2-CdA treatment. PMID- 9355025 TI - Surgical management of mycosis fungoides. AB - Mycosis fungoides (MF) is a type of cutaneous hyperproliferative T-cell disorder that may be localized. Although there is considerable controversy regarding whether MF may originate as a non-neoplastic condition, or even whether MF is a neoplastic condition until late in its course, we have seen a few cases undergo what appeared to us to be a clear progression from an inflammatory disorder to MF. We now report a 32-year-old man with MF most prominent on his right flank and buttock who developed his patches several weeks following, and in the precise locations in which he had experienced, exposure to toxic chemicals in an industrial accident. Because of this history, and because all lesions were transient except for these sites, these permanent lesions were treated with local surgical excision. There was no recurrence of disease at the treated sites, and the progression of MF markedly slowed following surgery, although he has continued to experience multi-focal transient recurrent disease, controlled by a combination of topical and systemic treatments, until the present time. Destructive methods such as excisional surgery or carbon dioxide laser may be considered a therapeutic option for localized MF. PMID- 9355026 TI - Magnetic resonance evaluation of uterine malformation with corpus agenesis. AB - Because no treatment is available and its rare prevalence, uterine fundal agenesis is seldom mentioned in the literature. A woman with uterine corpus agenesis, without other genital tract anomalies, presented with primary amenorrhea and infertility. Gonadotropins and ovarian steroids showed normal cyclic variations. A thorough evaluation, including history, physical examination, and appropriate imaging techniques [hysterosalpingography, ultrasonography and magnetic resonance imaging (MRI)] demonstrated extreme accuracy with MRI in the diagnosis of the anatomic defect. MRI permits noninvasive differentiation of uterine anomalies and may spare the patient's diagnostic laparoscopy. PMID- 9355027 TI - Signal averaged electrocardiography (SAECG) in patients on hemodialysis. AB - Cardiovascular diseases are the major cause of mortality in patients on hemodialysis (HD). Recently, signal averaged electrocardiography (SAECG) has been developed to detect ventricular late potentials (LP) noninvasively from the body surface for identifying patients at sudden death or ventricular tachycardia. We performed SAECG in 42 patients before and after HD. As a result, postdialysis total filtered QRS duration (FQRS) was significantly increased compared with predialysis FQRS. Postdialysis duration of low amplitude signal under 40 microV in the latter part of QRS (LAS40) tended to increase compared with predialysis LAS40. Before HD, there were no patients with LP and only one patient (2.4%) with abnormal SAECGs. In contrast, after HD, there were three patients (7.1%) with LP and three more patients (7.1%) with abnormal SAECGs. Furthermore, there was a significant correlation between the changes in LAS40 (delta LAS40) and those in potassium (K) (delta K) during HD. We further examined the relation between LAS40 and the concentration of K, by comparing the correlation coefficient between patients in the high-K group (predialysis K > or = 5.0 mEq/L; 20 patients) and those in the low-K group (predialysis K < 5.0 mEq/L; 22 patients). In the low-K group, there was no significant correlation between delta LAS40 and delta K. However, in the high-K group, there was a significant correlation between delta LAS40 and delta K. In conclusion, SAECG indices worsened during HD, and an insufficient decrement of serum potassium by HD is suggested to have been an arrhythmogenic factor in the high-K group. PMID- 9355028 TI - Anemia and thrombocytopenia due to parvovirus B-19 infection in a pregnant woman. AB - Anemia and thrombocytopenia in a patient with parvovirus B-19 and hepatitis C infection is reported. A seven month-pregnant 20 year-old patient had been first found to be anemic and thrombocytopenic 40 days before admission to our hospital and she had been given methylprednisolone and red cell transfusions. She gave birth to a healthy baby after only eight months of pregnancy. Ten days after delivery she was admitted to our hospital because of anemia and thrombocytopenia which did not respond to treatment. On admission, the blood count showed hemoglobin 8.1 g/dL, hematocrit 23.7%, white blood cells 11,200/microL, platelets 1000/microL, and reticulocytes 0.6%. Bone marrow smear and biopsy revealed erythroblastopenia and the absence of megakaryocytes. Liver enzymes were also high (alanine aminotransferase 1469 Units/L and aspartate aminotransferase 981 Units/L). In serologic studies PVB-19 IgM was found to be positive and hepatitis C virus RNA was detected. Red cell and platelet values returned to normal levels after cessation of methylprednisolone and concomitantly PVB-19 IgG was found positive in association with IgM in repeated determinations. PVB-19 was thought to be responsible for both anemia and thrombocytopenia. PMID- 9355029 TI - Apolipoproteins in obesity: effect of weight loss. AB - We evaluated serum concentrations of apoprotein (APO) A1, B, total cholesterol, triglycerides, high density cholesterol (HDL-C), and low density cholesterol (LDL C) in twelve obese subjects whose body mass index (BMI) was > or = 30 before and after a clinically significant weight loss was obtained utilizing a very-low calorie diet (VLCD) consisting of liquid protein (Optifast) providing 800 calories a day. At baseline, the mean weight +/- SD was 119.77 Kg and decreased significantly to 89.29 +/- 13.46 Kg by 24 weeks. Statistically significant reductions of APO-A1, APO-B, total cholesterol, and triglyceride concentrations were also observed along with the weight loss. LDL-C decreased from 156.0 +/- 55.9 mg/dL to 122.5 +/- 42.2 mg/dL (4.03 +/- 1.4 to 3.16 +/- 1.1 mmol/L), but this difference was not statistically significant. There was no significant change in the HDL-C and the ratios of APO-A1 to APO-B. We conclude that the use of VLCD is associated with changes in the lipid pattern that lower the cardiovascular risk profile in addition to the beneficial effects of weight loss itself. PMID- 9355030 TI - Eosinophilic gastroenteritis presenting with acute pancreatitis. AB - We present the findings on a 27-year-old male with eosinophilic gastro enterocolitis accompanied with acute pancreatitis. Acute pancreatitis may be induced by pancreatic duct obstruction caused by marked swelling of the papillary region of the duodenum due to eosinophil infiltration. After prednisolone treatment, clinical manifestations rapidly improved; the serum amylase decreased and the peripheral eosinophilia was recovered. The serum interleukin-5 (IL-5) level was high at diagnosis and decreased by prednisolone therapy; however, IL-5 was detected by enzyme immunoassay even during clinical remission. These results indicate that eosinophilia is mediated by IL-5, and detectable levels of IL-5 indicate the possibility of relapse. PMID- 9355031 TI - Nitric oxide is involved in the genesis of pulsatile LHRH secretion from immortalized LHRH neurons. AB - Neuronal networks controlling endocrine events present synchronous activity which is required for maintaining physiological functions, including reproduction. Although pulsatile hormone secretion is of paramount importance, the mechanism(s) by which secretory episodes are generated remain largely unknown. Nitric oxide (NO) has become the prototype of a new family of signaling molecules in the body. Nitric oxide diffuses from the source cell and controls activity of neighboring neurons as well as itself as a retrograde messenger. Cells of the luteinizing hormone-releasing hormone (LHRH) neuronal network, the key component in the control of reproduction, are scattered and loosely arranged in the anterior hypothalamus. A diffusible neurotransmitter could provide a means for establishing synchronous activation of the LHRH neuronal network leading to physiologically-relevant pulsatile LHRH secretion. In this study, we demonstrate that immortalized LHRH-producing neurons (GT1-7 cells) express NO synthase (NOS) mRNA and protein. Furthermore, GT1-7 cells are NADPH-diaphorase-positive (a marker of NOS activity) and the histochemical reaction can be abolished by treatment with a competitive NOS blocker. The presence of citrulline in these cells provides further evidence for the biological activity of NOS. These observations indicate that an active NO synthesizing machinery is present in immortalized LHRH neurons. In addition, we show that LHRH secretion is enhanced by NO in a cGMP-dependent manner. Since pulsatile LHRH secretion from immortalized LHRH neurons in vitro is abolished by NOS blockers and NO scavengers, it appears that NO is a unique neurotransmitter that is necessary to set LHRH neurons in phase to establish synchronized pulsatile LHRH secretion. PMID- 9355032 TI - Production of an oxytocin like substance by the subcommissural organ (SCO), related to the reproductive cycle in oviparous and viviparous reptiles. AB - The subcommissural organ (SCO) is a circumventricular organ of glial origin typical of all vertebrates. The SCO releases its secretion into the third ventricle to constitute Reissner's fibre (RF). Reportedly, in reptiles, SCO has cyclic secretory activity related to the reproductive cycle. In this immunocytochemical study we show that, in females of oviparous reptiles (Lacertidae: Podarcis sicula) and in a viviparous species (Scincidae: Chalcides chalcides), SCO secretion consists of hormones, in part of the oxytocin-like (OXY like) type. The amount of OXY-like material in the cells and in the third ventricle varies according to the different stages of the reproductive cycle. In the oviparous species, OXY-like hormone secretion can be induced by FSH administration at 28 degrees C, in the period of winter reproductive stasis as well. In the viviparous skink, showing an annual single ovulatory cycle, OXY-like secretion is present in the basal region of the cells, and is released into the third ventricle only at delivery. The role of an OXY-like hormone in the SCO is here discussed in relation to the different stages of the reproductive cycle. Its influence on the hypothalamus-hypophysis-gonad axis and its role in the transport of eggs into the ducts in the oviparous species, and at delivery in the viviparous one, are also suggested. PMID- 9355033 TI - Cortisol differentially regulates pituitary-adrenal function in the sheep fetus after disconnection of the hypothalamus and pituitary. AB - We have investigated the effects of a 5 day infusion of cortisol into fetal sheep, in which the hypothalamus and pituitary were surgically disconnected (HPD), on fetal pituitary-adrenal function. Fetal HPD and vascular catheterization were carried out at between 104 and 124 days gestation. Cortisol was administered (3.5 mg 24 h-1) for 120 h between 134 and 140 days (HPD + F group; n = 5) and saline was administered during the same gestational age range to HPD (HPD group; n = 12) and intact fetal sheep (Intact group; n = 6). Cortisol infusion into the HPD fetal sheep did not suppress the mRNA levels for Proopiomelanocortin (POMC) in the fetal anterior pituitary at 139/140 days gestation (POMC mRNA: 18S rRNA: Intact 0.40 +/- 0.05; HPD 0.56 +/- 0.07; HPD + F 0.49 +/- 0.07). Similarly, there was no significant effect of either HPD or cortisol infusion on the plasma concentrations of immunoreactive (ir) ACTH or ACTH(1-39). The adrenal: fetal body weight ratio was significantly higher, however, in the HPD + F (88.4 +/- 8.7 mg kg-1) and Intact groups (84.1 +/- 5.6 mg kg-1) when compared with the HPD fetal sheep (63.7 +/- 5.4 mg kg-1). The ratio of total IGF-II mRNA: 18S rRNA was similar in the adrenals of the Intact (0.48 +/- 0.09), HPD (0.78 +/- 0.09) and HPD + F (0.71 +/- 0.11) groups. The ratios of CYPIIA1, 3 beta-HSD and CYP21A1 mRNA: 18S rRNA were significantly lower in adrenals from the HPD group when compared to those in the Intact group and were not restored to normal by cortisol infusion. We have therefore demonstrated that cortisol does not act directly at the fetal pituitary to suppress POMC synthesis or ACTH secretion in late gestation. Cortisol does, however, stimulate fetal adrenal growth after HPD in the absence of any effects on adrenal IGF-II or steroidogenic enzyme mRNA levels. The data provide evidence that an intact hypothalamic-pituitary axis and cortisol each play an important role in the stimulation of adrenal growth and steroidogenesis which occurs during the last 10 15 days of gestation in the sheep. PMID- 9355034 TI - Primary cell culture of LHRH neurones from embryonic olfactory placode in the sheep (Ovis aries). AB - The aim of this study was to establish an in vitro model of ovine luteinizing hormone-releasing hormone (LHRH) neurones. Olfactory placodes from 26 day-old sheep embryos (E26) were used for explant culture. Cultures were maintained successfully up to 35 days, but were usually used at 17 days for immunocytochemistry. LHRH and neuronal markers such as neurofilament (NF) were detected by immunocytochemistry within and/or outside the explant. Three main types of LHRH positive cells are described: (1) neuroblastic LHRH and NF immunoreactive cells with round cell body and very short neurites found mainly within the explant, (2) migrating LHRH bipolar neurones with an fusiform cell body, found outside the explant, (3) network LHRH neuron, bipolar or multipolar with long neurites connecting other LHRH neurons. Cell morphology was very similar to that which has been described in the adult sheep brain. These results strongly suggest that LHRH neurones in the sheep originate from the olfactory placode. This mode may represent a useful tool to study LHRH neurones directly in the sheep. PMID- 9355035 TI - Microinjection of the tachykinin neuropeptide K into the ventromedial hypothalamus disrupts the hormonal onset of maternal behavior in female rats. AB - The medial amygdala exerts an inhibition of maternal behavior in virgin rats, but neither the site to which it projects to exert this effect nor the neurotransmitter used in such a pathway is known. There is also evidence that the ventromedial nucleus of the hypothalamus exerts an inhibition of maternal behavior, and the medial amygdala projects to this nucleus, suggesting that it may receive a projection from the medial amygdala which is inhibitory for maternal behavior. Tachykinin injection into the hypothalamus inhibits reproductive behavior in male rats, and there is a tachykininergic projection from the medial amygdala to the ventromedial hypothalamus. Consequently, the present study was conducted to evaluate the hypothesis that the tachykinin neuropeptide K can inhibit maternal behavior after injection into the ventromedial hypothalamus. Female rats were primed to be maternal by pregnancy termination and estrogen injection. Four doses of the peptide (279 pmol, 186 pmol, 116 pmol, and 66 pmol) were bilaterally injected into the ventromedial hypothalamus, and all were effective in delaying the onset of maternal behavior. Evidence that neuropeptide K disrupts maternal behavior in animals that have already begun to be maternal is also presented. Site specificity of neuropeptide K's effect to within the region of the ventromedial hypothalamus was supported, as injection into the mediodorsal thalamus was without effect. Some possibly relevant neuroendocrine effects are addressed, as well as the possibility that tachykinins may act within the ventromedial hypothalamus to promote virgin female rats' fear of pup odors. PMID- 9355037 TI - Castration decreases single cell levels of mRNA encoding glutamic acid decarboxylase in the diagonal band of broca and the sexually dimorphic nucleus of the preoptic area. AB - Using quantitative in situ hybridization histochemistry (ISHH), we determined the effect of castration on single cell levels of glutamic acid decarboxylase (GAD) mRNA in discrete hypothalamic regions of the male rat brain associated with the control of gonadotropin secretion. A 48-base oligodeoxynucleotide probe was used to detect with equal affinity the two isoforms of GAD message, GAD65 and GAD67. GAD message also was quantitated in a number of selected areas of the brain to contrast GAD gene expression amongst several populations of GABAergic neurons. Comparison of 11 brain regions demonstrated a 9.3-fold range in the quantity of single cell GAD mRNA with levels being highest in the amygdala and the diagonal band of Broca, moderate in the piriform cortex, caudate nucleus, substantia innominata, globus pallidus, cingulate cortex and medial septal nucleus, and lowest in the lateral septal nucleus and the medial preoptic nucleus (MPN). Castration markedly reduced single cell GAD mRNA levels in the DBB and the MPN, two discrete hypothalamic structures known to contain dendritic fields, cell bodies, and axons of GnRH neurons projecting to the median eminence. A striking finding was a dense core of steroid-sensitive GABAergic neurons within the MPN comprising the sexually dimorphic nucleus of the preoptic area (SDN-POA). Similar to the MPN as a whole, the amount of GAD mRNA expressed by cells in the SDN-POA of sham operated control rats was greater than in castrated animals. GAD mRNA levels were inversely related to serum LH titers, suggesting a role for these neurons in the mechanism controlling gonadal steroid negative feedback on LH secretion. This report provides the basis for future work to determine if GAD65, GAD67 or whether both isoforms are affected by gonadal steroid input. PMID- 9355036 TI - Regulation of pituitary corticotropin releasing hormone (CRH) receptor mRNA and CRH binding during adrenalectomy: role of glucocorticoids and hypothalamic factors. AB - The role of glucocorticoids and hypothalamic factors on CRH receptor expression in the pituitary were studied by analysis of the effects of adrenalectomy and suppression of CRH and VP secretion by hypothalamic lesions in the rat. Consistent with previous in situ hybridization studies, Northern blots showed that pituitary CRH receptor mRNA decreased only transiently after adrenalectomy, falling to 51% of the control levels after 18 h, and returning to control values after 6 days (112%). The early decrease was prevented by dexamethasone injection, 100 micrograms, s.c. The role of increased levels of CRH and VP in the pituitary portal circulation on the transient decrease in CRH receptor mRNA levels after adrenalectomy were studied by in situ hybridization in rats subjected in PVN lesions or median eminence deafferentation by hypothalamic anterolateral cuts (ALC). PVN lesion (12 days) or ALC (8 days) resulted in undetectable irCRH and VP in the external zone of the median eminence and had no effect on basal levels of pituitary POMC mRNA, CRH binding and CRH receptor mRNA. In sham lesioned rats, adrenalectomy for 18 h or 4 days caused the expected increases in pituitary POMC hnRNA and mRNA, and decreases in CRH binding. CRH-R mRNA levels decreased by about 50% after 18 h adrenalectomy but returned to basal by 4 days. PVN lesion or ALC fully prevented the fall in CRH binding after 18 h or 4 days adrenalectomy and the increase in POMC mRNA after 4 days adrenalectomy, whereas only attenuated the decrease in CRH receptor mRNA and increase in POMC mRNA levels after 18 h adrenalectomy. Administration of a CRH antagonist did not affect CRH receptor mRNA and POMC hnRNA and mRNA indicating that residual CRH in the median eminence after hypothalamic surgery is not responsible for the effect of adrenalectomy. These studies confirm previous in situ hybridization studies showing that adrenalectomy causes transient decreases in pituitary CRH receptor mRNA levels. The data indicate that while increases in hypothalamic CRH secretion following glucocorticoid withdrawal mediate pituitary CRH receptor binding loss and the increase in POMC expression after long-term adrenalectomy, CRH only partially accounts for the early changes in CRH receptor mRNA and POMC mRNA. PMID- 9355038 TI - Peptidases that degrade gonadotropin-releasing hormone: influence on LH secretion in the ewe. AB - Several peptidases have been postulated to degrade the hypothalamic peptide gonadotropin-releasing hormone (GnRH), but it is not known if such enzymes contribute significantly to the delivery of GnRH to the pituitary in vivo. Furthermore, the activity of GnRH-inactivating peptidases may vary in different reproductive states, such as across the estrous cycle. In the present study, specific fluorescent substrates were used to measure the activity of the two major GnRH-degrading enzymes, prolyl endopeptidase (PEP) and endopeptidase 3.4.24.15 (EP 24.15), in soluble extracts of the median eminentes (ME) of ewes during different phases of the estrous cycle. Levels of EP 24.15 and PEP activity in the ME did not vary significantly across the cycle, although PEP activity was lowest at the time of the preovulatory luteinizing hormone (LH) surge. However, a statistically significant decline in PEP activity (18%, P = 0.02) was observed in the ME of OVX ewes in which a surge was induced by estrogen when compared to oil treated OVX controls, suggesting a possible negative regulation of PEP activity by this steroid. The effect of intracerebroventricular (i.c.v.) infusion of several peptidase inhibitors on the pulsatile release of LH in the conscious OVX ewe was also examined. No consistent changes in the pattern of LH release were observed with i.c.v. infusion of the EP 24.15 inhibitor N-[1(R,S)-carboxy-3 phenylpropyl]-Ala-Ala-Tyr-p-aminobenzoate (cFP-AAY-pAB) or the angiotensin converting enzyme (ACE) inhibitor captopril. Similarly, administration of the prolyl endopeptidase inhibitor bacitracin, or a more specific inhibitor of this enzyme, Z-Proprolinal (ZPP), did not alter LH release patterns. The results did not demonstrate a major role for changes in the activity of EP 24.15, PEP, or ACE in altering the pattern of GnRH secretion, but a minor reduction in PEP levels may occur at the time of the estrogen-induced LH surge. PMID- 9355040 TI - Rapid changes of heteronuclear RNA for arginine vasopressin but not for corticotropin releasing hormone in response to acute corticosterone administration. AB - Corticotropin-releasing hormone and arginine vasopressin gene transcription were studied in the parvocellular cells of the hypothalamic paraventricular nucleus in response to both adrenalectomy and the injection of corticosterone. Following 6 days ADX, there was an increase in AVP mRNA, AVP hnRNA and CRH mRNA but not CRH hnRNA. After injection of corticosterone in ADX rats there was an extremely rapid reduction in AVP hnRNA which fell markedly within 15 min. AVP mRNA, however, remained raised throughout the 4 h. CRH hnRNA fell at 2 and 4 h, while CRH mRNA was only significantly reduced at the 4 h time point. Data demonstrate markedly different kinetics for the glucocorticoid regulation of the AVP and CRH genes in the hypothalamus. PMID- 9355039 TI - Effect of corticosterone and adrenalectomy on NMDA-induced cholinergic cell death in rat magnocellular nucleus basalis. AB - The present study demonstrates the effects of adrenalectomy and subcutaneously administered corticosterone on N-methyl-D-aspartate-induced neurodegeneration in the cholinergic magnocellular basal nucleus of the rat. NMDA was unilaterally injected into the nucleus basalis at different plasma corticosterone concentrations in adrenalectomized rats, in adrenalectomized animals with subcutaneously implanted cholesterol-corticosterone pellets containing 25% or 100% corticosterone, and in sham-adrenalectomized controls. The neurotoxic impact of the NMDA injection in the various experimental groups was assessed by the loss of cholinergic fibers stained with acetylcholinesterase histochemistry in the parietal neocortex. Reactive cortical astrocytes as a result of the treatments were detected by glial fibrillary acidic protein immunohistochemistry. Measurements of the densities of astrocytes and cholinergic fibers at the injected side of the brain were carried out by image analysis. Adrenalectomy significantly potentiated the NMDA-induced neurodegeneration by 50%, while chronic administration of corticosterone significantly attenuated the NMDA neurotoxicity in a dose-dependent manner. Compared to the ADX group, 25% corticosterone application reduced the NMDA damage by 37%, whereas the 100% corticosterone pellet diminished NMDA neurotoxicity by 75%. Both ADX and ADX + corticosterone implantation enhanced the NMDA-induced GFAP immunoreactivity. The increase of GFAP immunoreactivity was most pronounced in the adrenalectomized rats supplied with the 100% corticosterone pellets. The results demonstrate that corticosterone exerts a potent neuroprotective effect on NMDA-induced neurotoxicity in the magnocellular nucleus basalis. The activated astroglia suggest that astrocytes may contribute to the beneficial effect of corticosterone in the neuroprotective mechanisms against excitotoxic neuronal injury. PMID- 9355041 TI - Corticosteroid effects on gene expression of myelin basic protein in oligodendrocytes and of glial fibrillary acidic protein in type 1 astrocytes. AB - The paper describes the effects of corticosterone and deoxycorticosterone (DOC), used in their native or in their 5 alpha-reduced molecular forms (dihydrocorticosterone, DHC; dihydrodeoxycorticosterone, DHDOC; and tetrahydrodeoxycorticosterone, THDOC) on the gene expression of the myelin basic protein (MBP) and of the glial fibrillary acidic protein (GFAP) in pure cultures, respectively, of oligodendrocytes and type 1 astrocytes obtained from the neonatal rat brain. Among the different steroids tested (corticosterone, DHC, DOC, DHDOC and THDOC), only DHDOC was effective on the gene expression of MBP in the oligodendrocyte cultures; the mRNA levels of this typical oligodendrocyte marker were decreased following exposure to this steroid for 24 h. In the case of the astrocytic marker GFAP, its gene expression was increased by the exposure to corticosterone for 6 and 24 h, while DHC was ineffective; the mineralocorticoid DOC was also ineffective, while its 5 alpha-reduced derivative, DHDOC, strongly inhibited GFAP gene expression, starting at 6 h after beginning of the treatment. In conclusion, the present data show that: (1) adrenal steroids possessing gluco- and mineralocorticoid activities may influence the gene expression of the astrocytic marker GFAP; (2) the 5 alpha-reduced metabolite of DOC, DHDOC is able to influence the gene expression not only of GFAP but also that of MBP, which are, respectively, typical markers of the astrocytes and the oligodendrocytes; (3) the metabolic conversion of hormonal steroids into their 5 alpha-reduced metabolites, which also occurs in the glia, could be implicated in the biochemical control of oligodendrocyte and astrocyte functions. PMID- 9355042 TI - Response of substance P-immunoreactive nerve fibres in the anterior pituitary to plasma oestrogen levels in the rat. AB - The mammalian anterior pituitary has recently been shown to be innervated by substantial amounts of peptidergic nerve fibres. It has also been found that adrenalectomy increases the amount of calcitonin gene-related peptide-like immunoreactive nerve fibres in the anterior pituitary of the rat, and that stimulation of the nerve fibres in vitro can enhance release of ACTH. In the present study, the density of substance P-like immunoreactive nerve fibres in the anterior pituitary of the rat were examined in relation to variations in plasma oestrogen status. The amount of substance P-like immunoreactive nerve fibres increased significantly (P < 0.001) after ovariectomy, and decreased significantly (P < 0.01) under oestrogen replacement, in a dose-dependent manner. The results suggest the possibility of the direct neural involvement of oestrogen secretion in anterior pituitary regulation, which further substantiates the hypothesis of neural-humoral dual regulation of the mammalian anterior pituitary. PMID- 9355043 TI - Distinct agonist-mediated endocytosis of cloned rat somatostatin receptor subtypes expressed in insulinoma cells. AB - Endocytosis of somatostatin receptors could regulate cellular responses to the two natural peptides, somatostatin-14 and somatostatin-28, and to synthetic ligands used in the clinical diagnosis and symptomatic therapy of neuroendocrine tumours. The five cloned SSTRs with or without epitope tags at their carboxyl termini were expressed in rat insulinoma 1046-38 cells. Application of the two natural peptides or octreotide, at 37 degrees C but not at 4 degrees C, to cells transfected with somatostatin receptor subtype 2 or 3 cDNA resulted in a significant decrease of cell surface binding-sites for 125I-Tyr11-somatostatin 14. In contrast, cells transfected with subtype 5 cDNA only responded to stimulation with octreotide or somatostatin-28. Cells transfected with subtype 1 cDNA responded to somatostatin-14 and 28, while cells expressing subtype 4 cDNA showed no response. Confocal microscopy revealed that 6 min after stimulation with somatostatin-14 at 37 degrees C, tagged somatostatin receptor subtypes 1, 2 and 3 were internalized into vesicles. Internalization was not observed at 4 degrees C in the presence of 0.4 M sucrose and 80 microM phenylarsine oxide and hence proceeded via endocytosis through clathrin-coated pits and vesicles. After 20 min the internalized receptors appeared in perinuclear vesicles and after 120 min they reappeared at the plasma membrane. This recycling was not sensitive to cycloheximide and, hence, not dependent on de novo protein synthesis. Recovery of cell surface receptors was, however, inhibited by brefeldin A, monensin and bafilomycin A1, indicating that receptor recycling proceeded through vesicular traffic of acidified compartments. The data are consistent with the assumption that the observed agonist and subtype specific internalization of somatostatin receptors in a neuroendocrine cell line may be important for tumour diagnosis and therapy and, thus, suggest a manifold control in cellular signalling. PMID- 9355044 TI - Ontogeny of glucocorticoid receptors in the hyperstriatum-hippocampus parahippocampal area and optic tectum of the embryonic chicken (Gallus domesticus) brain. AB - The importance of glucocorticoids and their perturbation during development is an active research area. Developmental insults, including direct and indirect consequences of exposure to drugs of abuse or withdrawal from them, may act upon or via the neuroendocrine axis of the pregnant experimental subject (e.g. rat) and/or directly upon the neuroendocrine axis of the embryo or fetus. The use of the domestic chicken embryo may constitute a good experimental subject for studying these effects in the absence of maternal influences. Thus, the pattern of brain glucocorticoid cytosolic receptors were characterized in an early developing brain region, the optic tectum (OT) and a later developing region with a different function, the hyperstriatum-hippocampus-parahippocampal (HHP) area, on embryonic days (E) 11, 15, 18 and on the day of hatching (HD). The influence of the glucocorticoid synthesis inhibitor metyrapone, injected into eggs on E14 and on E17, upon glucocorticoid receptors (on E15 and E18) was also studied to determine effects of a 'chemical adrenalectomy'. Receptors for this steroid are high on E11 and E15, decreasing as they approach the time of hatching, with the HHP generally showing greater numbers of specific binding sites for [3H] corticosterone (CORT). Although metyrapone treatment did not alter the apparent number of receptors on E15, on E18 it unmasked receptors otherwise occupied by endogenous ligand(s) and/or induced their synthesis, resulting in significantly more receptors identified with [3H]-CORT. Nevertheless, the HHP continued to display more of these receptors than the OT on E15 and E18 after injection of metyrapone. These observations are consistent with the hypotheses that the HHP of embryos of this species contains a higher density of glucocorticoid receptors than does the OT; that glucocorticoid receptor quantification is related to steroid synthesis inhibition in late embryonic development; and that neuroendocrine feedback control of serum glucocorticoids may become functional between E15 and E18. The results also suggest the use of this experimental approach for assessing the effects of developmental insults with drugs, other than metyrapone, as a marker for altered neuroendocrine development and/or function. PMID- 9355045 TI - Gonadectomy alters tyrosine hydroxylase and norepinephrine transporter mRNA levels in the locus coeruleus in rabbits. AB - The central noradrenergic system has a major regulatory role on gonadotropin releasing hormone/luteinizing hormone (GnRH/LH) secretion in rabbits. Exogenous administration of norepinephrine (NE) alters GnRH/LH release in a sex steroid dependent manner, i.e. NE stimulates GnRH/LH release in oestrogen-primed ovariectomized (OVX) animals but not in non-primed individuals. To investigate how gonadal steroids influence noradrenergic neuronal activities in the locus coeruleus (LC), mRNA levels of tyrosine hydroxylase (TH) and NE transporter (NET), two key factors regulating NE synthesis and uptake, were compared 3 weeks after gonadectomy (GDX). Intact male (n = 5) and female (n = 6) New Zealand White rabbits were sacrificed along with castrated males (n = 4) and OVX females (n = 5). The brainstem from each individual was sectioned and the LC was punched for detection of TH and NET mRNA levels using the ribonuclease protection assay (RPA). Trunk blood was collected to determine immunoactive serum LH values. Levels of LH were elevated in both males and females after GDX. Luteinizing hormone concentrations averaged 0.10 +/- 0.05 ng/ml in intact males vs 1.64 +/- 0.31 ng/ml in castrated males (P < 0.01) and 0.30 +/- 0.08 ng/ml in intact vs 9.80 +/- 3.50 ng/ml in OVX females (P < 0.05), respectively. Removal of the gonads also increased TH mRNA levels in the LC in both males and females. In intact males, TH mRNA levels were 0.796 +/- 0.181 pg/microgram total RNA, whereas in castrates mRNA levels averaged 1.667 +/- 0.345 pg/microgram total RNA (P < 0.05). In intact females, TH mRNA levels were 0.617 +/- 0.054 pg/microgram total RNA while the OVX group averaged 1.084 +/- 0.202 pg/microgram total RNA (P < 0.05). Similar increases in NET mRNA were noted after GDX in both sexes. In males, NET mRNA levels were 1.461 +/- 0.401 pg/microgram total RNA in intacts vs 3.666 +/- 0.649 pg/microgram total RNA in castrates (P < 0.05). In females, NET mRNA levels averaged 1.336 +/- 0.212 pg/microgram total RNA and 3.297 +/- 0.835 pg/microgram total RNA in the intact and OVX groups, respectively (P < 0.05). The data indicate that GDX enhances gene expression of both TH and NET. The results support the hypothesis that the feedback regulation of sex steroids on LH secretion in rabbits of both sexes involves transcriptional/translational processes of at least TH and NET in brainstem NE cells. PMID- 9355046 TI - Secretory and morphological heterogeneity of porcine somatotropes during postnatal development. AB - Previous studies from our laboratory have demonstrated that porcine somatotropes can be separated into two subpopulations of low (LD) and high density (HD) by centrifugation in a Percoll gradient. The two subsets are present throughout porcine postnatal growth, although their relative proportions vary with age. In prepubertal animals, HD cells exhibit higher secretory granule content and release more GH than LD cells under basal culture conditions. In the present study, we analysed the ultrastructure of separated LD and HD cells from neonate and mature female pigs, and quantified cell size as well as the relative abundance of several subcellular organelles on immunoidentified somatotropes. Subsequently, GH release under basal conditions was assessed for cultures of unseparated cells and also for LD and HD somatotropes obtained at different stages of postnatal development. Results from the morphometric study demonstrated that LD somatotropes were significantly smaller in size, contained less secretory granules and displayed a more developed endoplasmic reticulum than their HD counterparts, regardless of the age of the pituitary donors. In terms of secretory ability, a significant age-associated decrease in GH release was observed in monolayer cultures of unseparated cells from prepubertal and mature pigs compared to neonates. A similar decline in GH-releasing ability was detected for cultures of HD cells. For LD cells, GH secretion only decreased significantly in mature animals. In spite of the divergent pattern followed by both subpopulations during growth, HD somatotropes released significantly more GH than LD somatotropes at the three ages studied. Taken together, our findings demonstrate that the population of porcine somatotropes is mainly composed of two subtypes, LD and HD, which differ in density, morphology and basal secretory activity. These differences are essentially maintained during porcine postnatal development. The progressive reduction in the secretory capacity of HD and LD somatotropes, coupled to the decrease in the relative abundance reported for the HD subpopulation, provides the cytological basis for a better understanding of the decline in GH release associated with age in pigs. PMID- 9355047 TI - Mesotocin gene expression in the diencephalon of domestic fowl: cloning and sequencing of the MT cDNA and distribution of MT gene expressing neurons in the chicken hypothalamus. AB - This study focuses on the structure and expression of the mesotocin (MT) gene in the chicken hypothalamus. Using an anchored and nested RT-PCR strategy, combined with circular RACE-PCR, the full length sequence of the chicken MT cDNA was obtained. The cDNA and derived amino acid sequences conformed to the structure of the oxytocin-like gene family. However, unlike most mammalian species, there does not appear to be frequent gene conversion between the MT and AVT cDNA sequences. A single specific hybridization signal of 1.2 kb was detected by Southern analysis of chicken genomic DNA, indicating only a single gene copy in the chicken genome. Northern analysis of hypothalamic RNA revealed a single band at approximately 0.6 kb. Using the same probe for in situ hybridization histochemistry, MT-mRNA was demonstrated to be predominantly localized in the parvocellular, magnocellular and periventricular subgroups of the paraventricular nucleus and, when compared to the distribution of neurons containing arginine vasotocin (AVT)-mRNA in the same region, with far fewer neurons expressing the MT gene in the lateral subgroups. Only few and scattered neurons expressing the MT gene were found in the ventral and external subgroups of the supraoptic nucleus in which many neurons contain AVT transcripts, as demonstrated in consecutive sections. In all nuclei investigated, the intensity of AVT and MT hybridization signals per cell was approximately equal. No specific labelling for MT-mRNA was found in the bed nucleus of the stria terminalis, nor the nucleus accumbens. Using immunocytochemical detection of AVT and in situ hybridization for neurons expressing MT-mRNA, some neurons were found to contain both AVT and MT gene products in the paraventricular nucleus but not in the supraoptic nucleus. PMID- 9355048 TI - Metabolic and orexigenic effects of intracerebroventricular neuropeptide Y are attenuated by food deprivation. AB - Administration of neuropeptide Y (NPY) into the hypothalamus or cerebral ventricles has been shown to increase food intake, the secretion of hormones such as insulin, glucagon and corticosterone and to alter the metabolism of carbohydrate and lipids. It has been suggested that metabolic effects of hypothalamic NPY may contribute to fat accretion in some types of obesity and to the metabolic and behavioural adaptation to food deprivation. However, it is currently unknown if different nutritional states alter the responses to hypothalamic NPY. Consequently, we have compared the effects of NPY injected into the third ventricle (ICV) in the fed and overnight-fasted state on ingestive behaviour, on insulin, glucagon and corticosterone secretion before, and following, an IV glucose bolus (IVGTT) and on blood glucose following an intra arterial insulin bolus (ITT). Studies were performed on conscious, unrestrained adult female rats. In the fed state, 2 and 6 micrograms ICV NPY produced a potent orexigenic and dypsogenic effect. In the fasted state, the 2 micrograms dose had a dypsogenic effect, while only the 6 micrograms dose had a significant orexigenic effect. In the fed but not fasted state, 3 micrograms ICV NPY increased plasma glucagon and corticosterone levels and attenuated the decline in blood glucose during the ITT. By contrast, in both fed and fasted groups, 3 micrograms ICV NPY potentiated the insulin secretory responses during the IVGTT. We conclude that, apart from stimulating insulin secretion, the acute metabolic and orexigenic responses to ICV NPY in this study were substantially reduced or abolished by overnight fasting. Therefore, behavioural and metabolic responses to endogenous hypothalamic NPY may also be more significant in the fed than the fasted state. PMID- 9355049 TI - Inhibition of brain oestrogen receptors by RU 58668. AB - The purpose of this study was to determine if injections of the 11 beta substituted steroidal antioestrogen, RU 58668, would block two measures of oestrogen receptor action in ovariectomized adult female rats. Using an in vitro nuclear exchange assay, it was found that RU 58668 reduced cell nuclear [3H] oestradiol binding in brain tissue 24 h after it was injected. However, pituitary cell nuclear [3H]-oestradiol binding was significantly reduced just 2 h after the antioestrogen was injected. Our results demonstrate that RU 58668 can reach the brain following a subcutaneous injection, but that it needs more time to reach the brain than it does to reach the pituitary. Since the levels of hypothalamic and pituitary progestin receptors are known to be regulated by oestradiol, cytosolic [3H]-R5020 binding was used as an in vitro assay of endogenous oestrogen receptor action. RU 58668 blocked induction by oestradiol of cytosolic [3H]-R5020 binding in both the brain and pituitary 48 h after it was injected. In the absence of oestradiol, RU 58668 did not stimulate cell nuclear [3H] oestradiol binding or cytosolic [3H]-R5020 binding in either brain or pituitary. In conclusion, injections of RU 58668 blocked two measures of oestrogen receptor action in the brain and pituitary without showing oestrogenic activity itself. PMID- 9355050 TI - Spondylolisthesis in the elite football player: an epidemiologic study in the NCAA and NFL. AB - Although spondylolisthesis in and of itself is not a contraindication to participation or successful performance in football, having spondylolisthesis may well predispose to symptoms and be associated with a worse prognosis. The purpose of this study was to determine the reported prevalence, treatment approach, outcomes, and perceptions regarding prognosis of elite football players with spondylolisthesis by their National Collegiate Athletic Association (NCAA) and National Football League (NFL) team physicians. A questionnaire regarding the prevalence, treatment, results, and perceptions regarding prognosis related to spondylolisthesis in football players was submitted to each team orthopaedic surgeon of the 28 NFL and the Final Associated Press ranked top-25 NCAA Division I teams at the conclusion of the 1993-1994 season. All questionnaires were returned for review. The prevalence of players with known spondylolisthesis currently participating in elite football was 1% in both the NCAA and NFL. Fifty two percent of NCAA and 43% of NFL team physicians were aware of at least one athlete with spondylolisthesis currently playing. Only six college and two NFL team physicians were aware of athletes surgically treated for spondylolisthesis. Sixty-four percent of NFL team physicians and 36% of college team physicians believed that the presence of spondylolisthesis implies a poor prognosis. Ninety six percent of professional team physicians downgraded the rating of players with known spondylolisthesis before the NFL draft. PMID- 9355051 TI - Prevalence of perioperative complications after anterior spinal fusion for patients with idiopathic scoliosis. AB - Anterior spinal fusion (ASF) has been proven to improve curve correction, save motion segments, and decrease the rate of pseudarthrosis when compared with posterior spinal fusion alone. However, in patients with idiopathic scoliosis, the complication rate of the anterior approach to the spine using current techniques has only been scantly defined in the literature. This is a retrospective review of consecutive patients who underwent primary ASF for idiopathic scoliosis to determine the prevalence and types of complications specifically related to the anterior approach. All patients who underwent primary ASFs for idiopathic scoliosis done by one of two orthopaedic surgeons between October 1986 and July 1992 were reviewed. Adequate records were available for 98 of 103 patients. The average age at time of surgery was 22 years (range, 10-60 years). Complications were divided into three groups: major (resulting in permanent sequelae or necessitating a second major operation); minor (resulting in a prolonged hospital stay, necessitating a minor operation, and/or resulting in a significant temporary hardship or persistent minor problem); and insignificant (anything less than minor). One of 98 patients had a major complication (a pelvic deep venous thrombosis that required operative thrombectomy). Twenty-five of 98 patients had 28 complications classified as minor, and 28 of 98 patients had 30 complications classified as insignificant. Smoking was a significant risk factor for the development of minor complications. There was no statistically significant relationship between the development of complications and the degree of curve, the approach used, the procedure performed, or the performance of rib resections. The anterior approach to the spine in patients with idiopathic scoliosis in this series was very safe, with only one major complication in 98 patients. However, minor and insignificant complications were quite common, occurring in 45 of 98 patients (46%). Smoking was a significant risk factor for minor complications. PMID- 9355052 TI - Lumbar spine surgery in the obese patient. AB - A prospective study was performed in obese and nonobese patients undergoing lumbar spine surgery to report perioperative data and surgical outcomes. One hundred fifty-nine consecutive patients who underwent lumbar spine surgery by a single surgeon entered the study. Among 159 consecutive patients, 55 met the criteria for obesity (> 20% ideal body weight). The average weight was 226 lb in the obese group. There were 28 men and 27 women with ages ranging from 25 to 81 years, with a mean of 47.9 years. The diagnoses and procedures included 31% herniated nucleus pulposus with laminotomy and discectomy, 14% spinal stenosis with laminectomy, and 55% lumbar fusion procedures for other disorders. Twenty nine percent had previous lumbar surgery that had failed. Results indicated that there were no significant differences between the obese and control groups in terms of duration of surgery (3.8 versus 3.2 h), blood loss (723 versus 605 ml), and duration of hospital stay (5.6 versus 5.8 days). The clinical results were 24% excellent, 40% good, 27% fair, and 9% poor in the obese group; and 27% excellent, 37% good, 19% fair, and 17% poor in the control group. This study found no significant differences between obese and control patients relative to blood loss, operative time, hospital stay, rate of complications, and functional outcome in lumbar spine surgery. Patient selection continues to be the most important factor in terms of operative success. We believe that lumbar spine surgery, including fusion, should not be withheld from obese patients who present with proper indications for surgery and fail an appropriate course of conservative treatment. PMID- 9355054 TI - Interobserver variability in grading lumbar fusions. AB - Inter- and intraobserver variability in grading lumbar fusion status radiographically was assessed. The objective was to determine the interobserver variability and intraobserver reproducibility in the assessment of two level noninstrumented lumbar fusions. Fifty sets of radiographs with anteroposterior, left and right bending, and flexion-extension lateral views were assessed by six observers of varying experience and background, with fusion status graded. Kappa statistical analysis revealed only fair interobserver agreement in grading lumbar fusion status. Intraobserver reproducibility was higher in more experienced observers. The results indicate only fair reliability in terms of interobserver agreement to grading of lumbar fusion status. Variability in assessing lumbar fusion radiographically may explain some of the variability in fusion rates reported in the literature and poor correlation that can be seen between clinical outcome and radiologic outcome. PMID- 9355053 TI - Radiographic mensuration characteristics of the sagittal lumbar spine from a normal population with a method to synthesize prior studies of lordosis. AB - Standing lateral lumbar radiographs of 50 normal healthy subjects were retrospectively selected for evaluation of lumbar lordosis. The objective was to evaluate, in a normal population, global and segmental contributions to lordosis in the standing position, and to devise a method to compare the seemingly unrelated multitude of lordotic values in the literature. Because of a variety of positioning and measurement methods of lordosis in live subjects and cadavers, correlation of results is difficult. While often relying on simple pain questionnaires, studies of normal subjects rarely have complete medical history, physical, neurological, and orthopedic examinations. Standing lateral lumbar radiographs of 50 subjects, who had complete histories and normal examinations, were analyzed to determine overall lordosis, segmental contributions, and vertical sagittal alignment. Using posterior body tangents, the mean L1-L5 angle was -39.7 degrees, CobbT12-S1 = -65 degrees, Ferguson's sacral angle = 39 degrees, pelvic tilt angle was 49 degrees, and average RRAs (segmental angles) were RRAT12-L1 = -3.6 degrees, RRAL1-L2 = -4.1 degrees, RRAL2-L3 = -7.6 degrees, RRAL3-L4 = -11.7 degrees, RRAL4-L5 = -16.8 degrees, and RRAL5-S1 = -32.4 degrees. Using segmental rotation angles as a method to compare past and current literature, a normal standing lumbar lordosis of CobbT12-S1 = -61 degrees, range 55 degrees to -65 degrees, was determined with specific segmental angles. PMID- 9355056 TI - The association of trunk muscle cross-sectional area and magnetic resonance image parameters with isokinetic and psychophysical lifting strength and static back muscle endurance in men. AB - The relationship between trunk muscle morphology as measured on transverse magnetic resonance images and isokinetic lifting, psychophysical lifting, and static back muscle endurance testing was examined in 110 men, ages 35-67 years (mean, 48 years), who had been chosen based on their exposure to a wide variety of occupational and leisure-time physical activities. The computed T2-relaxation times and the T2-weighted and proton density-weighted signal intensities of the erector spinae, quadratus lumborum, and psoas major muscles had almost no association with any of the strength tests. The cross-sectional areas of the muscles had good correlations with isokinetic lifting strength (r = 0.46-0.53). They did not correlate well with psychophysical lifting and static back muscle endurance. Other characteristics or neurological or psychological factors may have more influence on those tests. PMID- 9355055 TI - Comparison of trunk strength measurements between two different isokinetic devices used at clinical settings. AB - Intradevice reliability of isokinetic trunk strength measurements has been studied frequently, but no evidence is available on interdevice reliability. This motivated the present study, in which two isokinetic devices, the Ariel 5000 and Lido Multi-Joint II, were compared in a sample of 41 subjects (20 healthy and 21 low back pain subjects). The measurements were made in a random order with both machines. The results showed that the two isokinetic machines gave quite different results in trunk flexion-extension strength measurements. A statistically significant difference was present in the average peak torques between the two devices, with the exception of flexion at low angular velocity (60 degrees/s), and the correlations between the two measurements were low. The results were assumed to be more of a reflection of the interdevice variations (hardware and software, attachment of the subject) than of learning effects or other phenomena. We conclude that isokinetic trunk-muscle strength test results with the Ariel and Lido are device specific, and one cannot automatically compare results obtained from different devices with each other. PMID- 9355057 TI - Serial gadolinium-enhanced MR imaging after lumbar disc resection: observation of the affected root. AB - A prospective study was undertaken to clarify the relationship between postoperative morphological/pathological changes in the affected root and the clinical developments after disc resection. Gadolinium-enhanced magnetic resonance (MR) imaging was performed at 1 week, 5 weeks, 3 months, and 6 months after surgery for 28 patients of 34 consecutive patients who underwent single level disc resection. Enhancement/thickening of the affected root was found to be 100%/89% at 1 week, 50%/57% at 3 months, and 32%/37% at 6 months after surgery. Patients with root enhancement and thickening at 3 and 6 months after surgery had less clinical improvement than patients without it. There was consistent correlation between postoperative clinical developments and nerve root enhancement/thickening in enhanced MR imaging. To use enhanced MR imaging as an evaluation tool after disc surgery might increase the diagnostic accuracy and reduce failed back surgery syndrome. PMID- 9355058 TI - Preliminary results of the use of a two-stage treadmill test as a clinical diagnostic tool in the differential diagnosis of lumbar spinal stenosis. AB - This study assesses the ability of a two-stage treadmill test to distinguish stenotic from nonstenotic subjects by capitalizing on the postural dependency of stenotic symptoms. Forty-five subjects (26 stenotic, 19 nonstenotic) participated. An earlier onset of symptoms with level walking (p = 0.0009), increased total walking time on an inclined treadmill (p = 0.014), and prolonged recovery time after level walking (p = 0.001) were significantly associated with stenosis. Only one of four self-reported postural variables were significantly associated with stenosis. Linear discriminant analysis performed using the treadmill variables resulted in the correct classification of 76.9 and 94.7% of stenotic and nonstenotic subjects, respectively. Likelihood ratios for all treadmill variables were > 2.50, and < 2.00 for all self-report variables. A two stage treadmill test may be useful in the differential diagnosis of lumbar stenosis, and clinical measurement of the postural nature of symptoms seems to be superior to subjects' self-reports. PMID- 9355059 TI - Conservative treatment of tuberculous spondylitis: a long-term follow-up study. AB - A retrospective follow-up study was performed on 40 patients, in which tuberculous spondylitis was treated conservatively between 1969 and 1985 with orthotic supports for an average of 16 months (range, 10-30 months) and with anti tuberculous agents. All had persistent back pain, but none had neurological deficits. The mean follow-up period was 17 years (range, 10-26 years). Diagnosis was confirmed histopathologically. The spinal segments involved ranged from T5 to L5. The kyphotic angle was calculated according to Cobb. At final follow-up, 22 patients were pain free, 11 had occasional pain, 6 complained of pain in the morning, and 1 had chronic pain and needed frequent analgesics. Solid bony union was found in 75% of patients. The kyphotic deformity occurred in the thoracic spine with a mean angle of 20 degrees (range, 13-28 degrees) and in the lumbar spine with a mean angle 12 degrees (range, 5-26 degrees). The long-term follow-up of conservative treatment showed only slightly increased kyphosis. Conservative treatment is an alternative to surgical intervention in cases with kyphosis < 35 degrees. PMID- 9355060 TI - Anterior cervical fusion using a modified tricortical bone graft: a radiographic analysis of outcome. AB - A technique was developed to resist graft extrusion by means of mechanical interlocking between graft and vertebrae. A 4-mm burr is used to create a transverse trough (mortise) across the posterior aspect of each endplate. A reversed tricortical graft is shaped to have a ridge (tenon) composed of cortical bone on the upper and lower surfaces of its posterior portion. The ridges lock into the troughs to form a double mortise and tenon joint. Of 117 discs (102 patients) treated, adequate radiographs were available of 106 discs (in 92 patients) of which 89% were fused at last follow-up. Radiographic analysis showed significant (p < 0.001) increases in disc height and extension immediately postoperatively and a return to the preoperative values by the time of fusion. This technique completely eliminated graft extrusion without any reduction in fusion rate compared with most other reported results with the standard tricortical technique. PMID- 9355061 TI - Assessment of primary and salvage lateral mass screw insertion torque in a cadaveric model. AB - This study was performed to determine the insertion torques of three types of lateral mass screws and to explore their relationship to bone mineral density (BMD). The peak insertion torque of primary articular screws was measured, and the holes were then stripped by overtightening. Larger diameter "salvage" screws were placed in the stripped holes, and peak insertion torque was measured. Peak primary screw insertion torque and BMD were positively correlated (r = 0.48, p = 0.00001). The ratio of salvage to primary screw insertion torque varied significantly among the systems (p = 0.0003). The positive correlation between BMD and primary screw peak insertion torque confirms the importance of bone density for satisfactory screw purchase. The large variation in the ratio of salvage to primary screw insertion torque among systems may be due to differences in primary and salvage screw design. PMID- 9355062 TI - Bivector traction for unstable cervical spine fractures: a description of its application and preliminary results. AB - The management of acute, displaced odontoid fractures requires the restoration of sagittal alignment and rigid external or internal immobilization to prevent late instability and achieve union. This report introduces a new traction technique for the reduction of posteriorly displaced type 2 odontoid fractures. Seven patients with traumatic injuries to the dens were placed in bivector traction for an awake closed reduction. Sagittal alignment was restored and maintained in all patients with no neurologic deterioration or traction-related complications during an average of 11 days (range, 2-28 days) in traction. The overall sagittal alignment corrected from an initial average of 12.2 mm (range, 5-22 mm) of posterior displacement to an average of 1.1 mm (range, 0-3 mm) at the completion of reduction. Only one patient had residual angulation, which measured 5 degrees. Three patients achieved an osseous union and the remaining four required a posterior C1-C2 fusion for nonunion. Although operative stabilization may be the preferred approach in this patient population and injury pattern, we conclude that bivector traction is a safe and effective technique for the initial management of posteriorly displaced odontoid fractures. In addition, its role can be expanded to the closed reduction of lower cervical spine fractures in patients with fixed flexion deformities secondary to ankylosing spondylitis or disseminated intraosseous segmental hyperostosis. PMID- 9355063 TI - Transient paralysis associated with epidural steroid injection. AB - Epidural steroid therapy is a commonly applied "conservative" therapy, but it is not inherently benign. Although arachnoiditis, infection, and meningitis have been reported, acute paraplegia has not been reported as a complication of either caudal or spinal epidural steroid injection. A unique case of transient, profound paralysis after epidural steroid injection is reported here. The procedure was carried out without fluoroscopic control and was complicated by a puncture of the thecal sack. Radiographic studies demonstrated a focal, space-occupying lesion in the spinal canal at the level corresponding to the neurologic deficit, which spontaneously resolved over the next 2-3 h. Surgical decompression was initially considered and then deferred in favor of observation. The patient recovered motor, sensory, and bowel and bladder function over the next 48 h. The period of recovery was consistent with an acute but brief compressive injury and inconsistent with an anesthetic effect. Radiographic studies suggest three possible explanations: (a) inadvertent thecal penetration during injection may have produced an atypical anesthetic block; (b) loculation of the injected fluid may have caused a transient compressive lesion; or (c) intrathecal injection may have produced an iatrogenic arachnoid cyst. Although pathologic confirmation of the cause was not possible, the potential for this alarming complication should be recognized by physicians prescribing epidural steroid therapy. We do not suggest that epidural steroid therapy is the treatment of choice for patients with multiple back operations or that it is efficacious for these patients. Our purpose is to alert surgeons and therapists to a rare but potentially devastating complication and to provide our experience in treating it. PMID- 9355064 TI - Displaced fracture in a hemangiomatous odontoid. AB - Vertebral hemangioma is a common and usually benign lesion encountered on routine radiographic studies. We report the unusual case of a woman who sustained a displaced fracture of a hemangiomatous odontoid after minimal trauma. Good healing was accomplished with external halo bracing alone. PMID- 9355065 TI - Foraminal herniation of a lumbar disc mimicking neurinoma on CT and MR imaging. AB - A case of a large L3-L4 intervertebral disc herniation causing a widening of the intervertebral foramen is reported. There was a soft-tissue mas within the spinal canal and intervertebral foramen. The soft-tissue mass within the spinal canal and intervertebral foramen showed marked enhancement on magnetic resonance scans after injection of a paramagnetic contrast agent (Gd-DTPA). The combination of lesion configuration and paramagnetic contrast enhancement mimicked a spinal neurinoma. There exists a degree of overlap between the imaging of herniated discs and spinal neurinomas. When this overlap involves several aspects, such as anatomic configuration, mass enhancement, and secondary foraminal dilatation, the differential diagnosis between a herniated disc and a neurinoma may be problematic. PMID- 9355066 TI - Traumatic bilateral rotatory dislocation of the atlanto-axial joints: a case report and review of the literature. AB - Traumatic bilateral rotatory dislocation at the atlanto-axial joints is a rare injury in adults. Only three prior cases have been reported (1,2,3). Our case report, review of management, and pathophysiology from the literature is presented. This injury may be successfully treated by closed reduction and brace immobilization. PMID- 9355067 TI - Epithelial galectin-3 during human nephrogenesis and childhood cystic diseases. AB - Galectin-3 is a beta-galactoside-binding protein with putative roles in development, oncogenesis, and inflammation. Its expression in human nephrogenesis has not been previously reported. This study examines galectin-3 expression in early human embryos by Western blot and immunohistochemistry. This 33-kD protein was detected in the apical domain of distal tubules of the mesonephros and also in the mesonephric duct. In the metanephros, the adult kidney precursor, galectin 3 was detected in the apical domains of ureteric bud branches, and there was intense expression in fetal medullary and papillary collecting ducts in both the cytoplasm and plasma membranes. Low levels of galectin-3 were detected in the cytoplasm of a subset of cells in adult collecting ducts; these were alpha intercalated cells because they expressed basal band 3 protein. In human multicystic dysplastic kidneys, all diseased epithelia had an embryonic apical expression pattern of galectin-3 and, in addition, all cystic epithelia in autosomal recessive polycystic kidneys expressed this molecule. It is concluded that galectin-3 is expressed by cells of the mesonephric duct/ureteric bud lineage, and it is speculated that the different subcellular locations may be implicated in both the regulation of normal growth and differentiation of this lineage, as well as in the pathogenesis of cystic epithelia. PMID- 9355068 TI - Expression of type 1 angiotensin II receptor subtypes and angiotensin II-induced calcium mobilization along the rat nephron. AB - The localization of two type 1 angiotensin II receptor subtype mRNA, AT1A and AT1B, was determined by reverse transcription-PCR on microdissected glomeruli and nephron segments. The coupling sensitivity of these two receptor subtypes was evaluated by measuring variations in intracellular calcium ([Ca2+]i) elicited by angiotensin II (Ang II) in structures expressing either AT1A or AT1B mRNA, using Fura-2 fluorescence. The highest expression of AT1 mRNA was found in glomerulus, proximal tubule, and thick ascending limb. In glomerulus, AT1A and AT1B mRNA were similarly expressed, whereas in all nephron segments AT1A mRNA expression was dominant (approximately 84%). The increase in [Ca2+]i elicited by 10(-7) mol/L Ang II was highest in proximal segments (delta [Ca2+]i is approximately equivalent to 300 to 400 nmol/L) and thick ascending limb (delta [Ca2+]i is approximately equivalent to 200 nmol/L). In glomerulus and collecting duct, the response was lower (delta < 100 nmol/L). The median effective concentrations for Ang II were of the same order of magnitude in glomerulus (12.2 nmol/L), in which both AT1A and AT1B are expressed, and in cortical thick ascending limb (10.3 nmol/ L), in which AT1A is almost exclusively expressed. The Ang II-induced calcium responses were totally abolished by the AT1 receptor antagonist losartan (1 mumol/L) but not by the AT2 antagonist PD 123319 (1 mumol/L). In the absence of external Ca2+, the peak phase of the response induced by 10(-7) mol/L Ang II was reduced and shortened, suggesting that a part of the [Ca2+]i increase originated from the mobilization of the intracellular Ca2+ pool. In conclusion, these results demonstrate that in the rat kidney: (1) AT1A is the predominant AT1 receptor subtype expressed in the nephron segments, (2) glomerulus is the only structure with a relatively high AT1B mRNA content, and (3) AT1A and AT1B receptor subtypes do not differ in their efficiency for the activation of calcium second-messenger system. PMID- 9355069 TI - Reversible glomerular hypertrophy in adult patients with primary focal segmental glomerulosclerosis. AB - The present study was performed to assess the pathogenetic role of glomerular hypertrophy in patients with primary focal segmental glomerulosclerosis (FSGS). We studied 14 patients with FSGS by morphometry. In seven patients, minimal change nephrotic syndrome (MCNS) was diagnosed on the first renal biopsy, but FSGS was diagnosed on the second biopsy (MCNS-FSGS group). Seven other patients with FSGS on the first biopsy underwent second biopsies while in remission (FSGS R group). Biopsy results were compared with biopsies from 10 patients with MCNS and seven control subjects. Nonsclerotic glomeruli were examined. The mean glomerular tuft area, whole glomerular area, and number of mesangial cells were significantly increased in both biopsies from the MCNS-FSGS group and in the first biopsies obtained during the nephrotic stage of the FSGS-R group, compared with control subjects and patients with MCNS. Biopsies from FSGS patients in remission showed that the mean glomerular tuft area and number of mesangial cells were significantly decreased. The fractional extracellular matrix area (extracellular matrix area/glomerular tuft area) and mesangial cell density (mesangial cell number/glomerular tuft area) in FSGS during both nephrotic and remission stages were the same as those in control subjects and patients with MCNS. The present study suggests that glomerular hypertrophy precedes the development of glomerulosclerosis in FSGS and is reversible when patients are in remission. These features support the pathogenetic importance of glomerular hypertrophy in patients with primary FSGS. PMID- 9355071 TI - Antiproliferative activity to glomerular mesangial cells and receptor binding of a heparin-mimicking polyaromatic anionic compound. AB - Proliferation of mesangial cells (MC) is a key feature in the pathogenesis of numerous renal diseases involving the glomerulus. Heparin, one of several compounds capable of suppressing MC proliferation, did not prove beneficial in the treatment of human glomerular diseases. In a search for a superior antiproliferative agent, a synthetic polyaromatic "heparin mimicking" compound (RG-13577, polymer of 4-hydroxyphenoxy acetic acid, M(r) approximately 5800), previously reported to inhibit the proliferation of vascular smooth muscle cells, was applied. RG-13577 exhibits approximately 1% of the anticoagulant activity of heparin and is nontoxic in animal experiments. Proliferation of primary rat MC was almost completely inhibited in the presence of 10 to 25 micrograms/ml RG 13577, and 50% inhibition was obtained at 1 to 5 micrograms/ml RG-13577. The cells resumed their normal growth rate after removal of RG-13577 from the culture medium. Under the same conditions, heparin exerted only a small inhibitors effect. RG-13577 inhibited signaling (i.e., tyrosine phosphorylation) and MC proliferation induced by both basic fibroblast growth factor and platelet-derived growth factor. RG-13577 binds to a naturally produced extracellular matrix, and the bound molecule retained its antiproliferative effect toward MC. 14C-Labeled RG-13577 also binds to cultured MC in a specific and saturable manner. Binding of 14C-RG-13577 was reduced by 80 to 90% in the presence of excess unlabeled RG 13577, apolipoprotein E, or lactoferrin, but there was no effect with heparin. Furthermore, the antiproliferative effect of RG-13577 was abolished in the presence of lactoferrin. It is proposed that compound RG-13577 inhibits MC proliferation through neutralization of growth-promoting factors, primarily heparin-binding growth factors, and possibly through binding to specific cell surface receptors, most likely the LDL receptor-related protein. RG-13577 and related polyanionic compounds may be applied to inhibit MC proliferation in glomerular diseases. PMID- 9355070 TI - Expression of beta 1-integrins on activated mesangial cells in human glomerulonephritis. AB - beta 1-integrins, a family of cell-surface receptors, mediate cell-matrix interactions that play a critical role in tissue development and tissue remodeling after injury. In this study, to clarify the importance of beta 1 integrins in human glomerulonephritis (GN), the relationship among the glomerular expression of beta 1-integrins, their ligand matrix components, alpha-smooth muscle actin (alpha-SM actin) as a marker of activated mesangial cells (MC), transforming growth factor-beta (TGF-beta), and glomerular cellularity in two normal kidneys, ten minimal change nephrotic syndrome, 23 immunoglobulin A (IgA) GN, 13 lupus GN, and four membranous GN kidneys were studied. Immunostaining was performed on frozen sections, using monoclonal anti-alpha-SM actin antibody and polyclonal antibodies against fibronectin, collagen type IV, laminin, each subunit of alpha 1 beta 1 (collagen/laminin receptor), alpha 5 beta 1 (fibronectin receptor) and TGF-beta. Quantitation of staining indicated that the glomerular expression of alpha 1 beta 1 and alpha 5 beta 1 integrins correlated with the mesangial amounts of their ligands, collagen type IV, laminin and fibronectin (P < 0.01), alpha-SM actin (P < 0.01), and TGF-beta (P < 0.01). In addition, a correlation was observed between an increased expression of alpha 1 beta 1 and alpha 5 beta 1 integrins and the degree of glomerular cell proliferation (P < 0.01). Double immunostaining showed that activated MC expressing alpha-SM actin strongly expressed alpha 1 beta 1 and alpha 5 beta 1 integrins, and these MC phenotypic alterations paralleled the level of glomerular TGF-beta staining (P < 0.01). In conclusion, enhanced expression of beta 1 integrins by activated MC may contribute to the pathological mesangial remodeling characterized by MC proliferation and matrix deposition in human GN. Increased glomerular TGF-beta appears to be involved in these MC phenotypic changes. PMID- 9355072 TI - Mapping B cell epitopes in Goodpasture's disease. AB - The target antigen of the pathogenic autoantibodies in Goodpasture's disease is the noncollagenous domain of the alpha 3 chain of type IV collagen. A panel of membrane-bound peptides was used to identify antibody-binding regions within the protein, and the significance of the binding by inhibition studies using soluble peptides, and by a structural analysis of the antigen, was confirmed. A total of 117 overlapping 12-mer peptides spanning the entire antigen was simultaneously synthesized as individual spots on a cellulose membrane. All nine patients' sera bound to the membrane, with a conserved pattern of peptides recognized by all sera. Inhibition studies were performed using a panel of overlapping 20-mer peptides, also spanning the entire antigen. Peptides from the regions that bound IgG were able to inhibit the binding of autoantibodies to native antigen. Predictions of the secondary structure of the noncollagenous domain of the alpha 3 chain of type IV collagen were performed by a conventional hydropathy plot and by multiple alignment of homologous alpha chains of type IV collagen and comparison with a structural data base. The core peptides binding and inhibiting Goodpasture's antibodies were predicted to be surface exposed and antigenic. Thus, the conformational epitope(s) of the Goodpasture antigen can be mapped using linear peptides. PMID- 9355073 TI - A common NKCC2 mutation in Costa Rican Bartter's syndrome patients: evidence for a founder effect. AB - Bartter's syndrome involves an overlapping set of closely related renal tubular disorders that can be subdivided into at least three clinical phenotypes: (1) the hypercalciuric antenatal Bartter variant; (2) the classic Bartter variant; and (3) the hypocalciuric-hypomagnesemic Gitelman variant. Recent data demonstrate that in several phenotypically indistinguishable cohorts, antenatal Bartter's syndrome is genetically heterogeneous. In these patients, mutations in the genes encoding either the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2) or the ATP-regulated potassium channel ROMK (KCNJI) have been identified. A cohort of 20 Costa Rican patients with a congenital syndrome that bears strong similarities to antenatal Bartter's syndrome but also has several distinct features has recently been described. In this cohort, we have identified a predominant mutation that introduces a premature stop in codon W625 of the NKCC2 gene (SCL12A1). This mutant allele is contained on a single common haplotype, suggesting that the majority of antenatal Bartter's syndrome patients in Costa Rica share a single common ancestor. PMID- 9355074 TI - Role of nitric oxide in rat nephrotoxic nephritis: comparison between inducible and constitutive nitric oxide synthase. AB - Nitric oxide (NO), generated by inducible NO synthase (iNOS) in migrating macrophages, is increased in glomerulonephritis. This study investigates the effect of NO inhibition on rat nephrotoxic nephritis (NTN) to clarify the role of NO production in glomerular damage. NTN was induced in Sprague Dawley rats by an injection of an anti-glomerular basement membrane (GBM) antibody. Urinary nitrite excretion and nitrite release from kidney slices (5.47 +/- 1.19 versus 2.15 +/- 0.73 nmol/mg protein, NTN versus Control, P < 0.05) were increased in NTN on day 2. Glomerular macrophage infiltration and intercellular adhesion molecule (ICAM) 1 expression increased from day 2. iNOS expression was increased in interstitial macrophages. Glomerular endothelial cell NOS (ecNOS) expression evaluated by counting immunogold particles along GBM was suppressed (0.06 +/- 0.02 versus 0.35 +/- 0.04 gold/micron GBM, P < 0.0001). Glomerular damage developed progressively. NG-nitro-L-arginine methyl ester (L-NAME), which inhibits both iNOS and ecNOS and aminoguanidine (AG), a relatively selective inhibitor for iNOS, equally suppressed nitrite in urine and renal tissue. Glomerular ICAM-1 expression and macrophage infiltration were reduced by L-NAME, but not by AG. Expression of ecNOS was significantly increased by L-NAME (0.91 +/- 0.08, P < 0.0001 versus NTN), but slightly by AG (0.18 +/- 0.04). AG significantly and L-NAME slightly attenuated the glomerular damage at day 4. In conclusion, suppression of iNOS prevents glomerular damage in the early stage of NTN. Treatment by L-NAME reduces macrophage infiltration by suppression of ICAM-1 expression, which may be explained by an increase in ecNOS expression. PMID- 9355075 TI - Reactive oxygen species and antioxidant defense in puromycin aminonucleoside glomerulopathy. AB - Results from several radical scavenger studies indirectly suggested an involvement of reactive oxygen species in the pathogenesis of puromycin aminonucleoside glomerulopathy. In this study, generation of reactive oxygen species was examined directly in glomeruli isolated from rats in the acute phase of puromycin aminonucleoside nephrosis and related to the changes in the glomerular antioxidant defense. Five and nine days after puromycin aminonucleoside injection, gross proteinuria, reduced creatinine clearances, and typical changes of glomerular morphology were present. Levels of reactive oxygen species were increased eightfold in glomeruli isolated 15 min after puromycin aminonucleoside injection, returned to baseline levels on days 1 and 5 after injection, and rose again to 14-fold on day 9 after injection, as determined by chemiluminescence with luminol. Further analysis of increased glomerular radical generation, using the chemiluminescence enhancer lucigenin and different radical scavengers, suggested a predominant involvement of hydroxyl radical and hydrogen peroxide in the initial increase in reactive oxygen species 15 min after puromycin aminonucleoside. Nine days after induction of nephrosis, primarily superoxide anion and hydroxyl radical were found to contribute to increased reactive oxygen species. Despite oxidative stress, antioxidant enzymes were not induced in the course of nephrosis. On the contrary, catalase and glutathione peroxidase activities declined 9 d after puromycin aminonucleoside injection. The results indicate that a transient increase in glomerular reactive oxygen species is sufficient to induce the oxidative glomerular injury observed in this model and that the glomerulus may not necessarily respond to oxidative stress with an induction of antioxidant enzymes. PMID- 9355077 TI - Fish oil therapy for IgA nephropathy: efficacy and interstudy variability. AB - Published reports examining the efficacy of fish oil for preserving renal function in immunoglobulin A (IgA) nephropathy have yielded conflicting results. This investigation was a meta-analysis conducted to determine whether the medical literature supports this therapy. In addition, the sources of variability among published findings were examined. Studies were combined using a random effects model. Five controlled studies were identified, two with positive results and three with negative results. Forty-four percent of the between-study variance could be attributed to differences in follow-up times and, less significantly, the number of renal function measurements; a weighting procedure was developed, eliminating this variance from the combined result. When all studies were combined, the mean effect, +0.25 +/- 0.23 SD (positive effects indicate that treatment was superior to control), was not statistically significant; however, the probability of at least a minor beneficial effect was 75%. Mixed-effects regression suggested that this therapy may be more effective among individuals with more proteinuria. The medical literature, therefore, does not prove the efficacy of fish oil therapy in IgA nephropathy, but suggests that an additional placebo-controlled trial is warranted. A sample-size calculation indicated that such a trial should be larger than those to date or should attempt to increase the treatment effect, perhaps by treating for more than 2 yr or enrolling more severely proteinuric individuals. PMID- 9355076 TI - Interaction of angiotensin II and TGF-beta 1 in the rat remnant kidney. AB - An interaction between angiotensin (Ang) II and transforming growth factor (TGF) beta 1 is gaining increasing recognition. Ang II has been implicated in the progression of renal disease, and TGF-beta 1 is a potent fibrosis-promoting cytokine. We sought to determine whether the beneficial effects of renin angiotensin system blockade on remnant kidney function were associated with a reduction in renal TGF-beta 1 in this model of chronic renal failure. After subtotal renal ablation, rats fed a 40% protein diet and treated with losartan not only had a reduction in systolic BP (96 +/- 8 versus 130 +/- 8 mmHg, P < 0.05, losartan versus control) and urinary protein excretion (4 +/- 5 versus 23 +/- 20 g/d, P < 0.05, losartan versus control), but also exhibited a reduction in renal TGF-beta 1 mRNA (194 +/- 64 versus 411 +/- 101 optical density units, P < 0.05, losartan versus control) and TGF-beta 1 protein levels (9.8 +/- 2.5 versus 18.6 +/- 5.8 ng/g of renal tissue, P < 0.05, losartan versus control). The elevation of TGF-beta 1 in the remnant kidney was most pronounced in the scar region (22.9 +/- 13.1 versus 5.8 +/- 3.7 ng/g, P < 0.05, scar versus nonscar). A combination of reserpine, hydralazine, and hydrochlorothiazide, although effective in lowering systemic BP in this model of chronic renal failure, was not associated with a reduction in proteinuria or TGF-beta 1. We conclude that in this model of progressive renal injury, Ang II antagonism may exert a beneficial effect in part by its negative influence on TGF-beta 1. PMID- 9355079 TI - An analysis of risk factors for withdrawal from dialysis before death. AB - Withdrawal from dialysis has been a significant cause of mortality among dialysis patients, accounting for 6 to 22% of deaths. Since 1990, a new death notification form has allowed more detailed analyses of withdrawal from dialysis separate from causes of death. Using the U.S. Renal Data System data base, this study examined 116,829 deaths in adult patients from 1990 to 1995. Adjusted odds ratios were calculated for the risk of withdrawal using logistic regression. Adjustments included age at death, ethnicity, gender, cause of death, primary cause of end stage renal disease, time on dialysis, and dialysis modality. In addition, odds ratios of withdrawal were calculated for deaths in patients who started dialysis after age 65. Death was preceded by withdrawal significantly more frequently in women than in men, more than twice as frequently in Caucasians than in African Americans or Asians, and more frequently in older than in younger age groups. Patients who died of chronic diseases (e.g., dementia, malignancy) were much more likely to withdraw before death, whereas patients who died from more acute causes (e.g., coronary artery disease) were less likely to withdraw before death. It is concluded that patients who are Caucasian, female, older, or die of chronic or progressive diseases are more likely to withdraw from dialysis before death. The ethnic and gender differences in withdrawal do not appear to have a medical explanation from this analysis. Further research along sociologic lines is needed to better explain the differences in withdrawal from chronic dialysis. PMID- 9355078 TI - Cytokine profiles during clinical high-flux dialysis: no evidence for cytokine generation by circulating monocytes. AB - Secretion of cytokines by monocytes has been implicated in the pathogenesis of dialysis-related morbidity. Cytokine generation is presumed to take place in two steps: induction of mRNA transcription for cytokines by C5a and direct membrane contact, followed by lipopolysaccharide (LPS)-induced translation of mRNA (priming/second signal theory, Kidney Int 37: 85-93, 1990). However, the in vitro conditions on which this theory was based differed markedly from clinical dialysis. To test this postulate for routine hemodialysis, 13 patients were studied cross-over with high-flux cuprammonium (CU), cellulose triacetate (CTA), and polysulfon dialyzers, using standard bicarbonate dialysate, as well as CTA with filtered dialysate (fCTA). Besides leukocytes, C3a, C5a, and limulus amebocyte lysate reactivity, tumor necrosis factor (TNF)-alpha, interleukin (IL) 1 beta, IL-6, IL-1RA, soluble TNF receptors, and IL-1 beta mRNA were assessed. Only during dialysis with CU did C5a increase significantly (561 to 8185 ng/ml, P < 0.001). Endotoxin content of standard bicarbonate was higher than filtered dialysate (median, 24.3 and < 5 pg/ml respectively, P = 0.002), whereas limulus amebocyte lysate reactivity was not detected in the blood, except in the case of CU. TNF-alpha levels were elevated before, and remained stable during, dialysis, independent of the modality used. IL-1 beta, IL-6, and mRNA coding for IL-1 beta could not be demonstrated. IL-1RA and soluble TNF receptors (p55/p75) were markedly elevated compared with normal control subjects, but showed no differences between fCTA and CTA. To summarize, no evidence was found for production and release of cytokines by monocytes during clinical high-flux bicarbonate hemodialysis, neither with complement-activating membranes nor with unfiltered dialysate. Therefore, this study sheds some doubt on the relevance of the "priming/second signal" theory for clinical practice. The data presented suggest that reluctance to prescribe the use of high-flux dialyzers, as advocated in many reports, may not be warranted. PMID- 9355080 TI - Relationship between left ventricular hypertrophy and plasma renin activity in chronic hemodialysis patients. AB - Left ventricular hypertrophy (LVH) is very common in uremic patients. It was shown previously that hemodialysis patients are chronically exposed to the extremes of plasma renin activity due to differences in the original renal disease. Because nonhemodynamic factors seem to play a fundamental role in the development of LVH, the present study was undertaken to investigate the relationship between the predialysis renin level and the echocardiographically determined cardiac structure in stable hemodialysis patients, matched for other parameters known to participate in the development of LVH, such as age; gender; body mass index; interdialytic weight gain; heart rate; systolic, diastolic, and mean arterial BP; hematologic and biochemical profile; vascular access; adequacy of dialysis; nutritional status; and period of follow-up. Thirty-three such patients were stratified in three groups according to predialysis renin levels: group A (n = 11), with renin levels < or = 1 ng.ml-1.h-1; group B (n = 9), with renin levels between 1 and 4 ng.ml-1.h-1; and group C (n = 13), with renin levels > or = 4 ng.ml-1.h-1. LVH with disproportionate septal thickening was directly related to the degree of renin-angiotensin system activation, and values for interventricular septum thickness, posterior wall thickness, interventricular septum thickness/posterior wall thickness ratio, left ventricular mass, and left ventricular mass index were all significantly correlated with predialysis renin levels. Because angiotensin II promotes growth in both fibroblasts and cardiac myocytes, these relationships suggest that elevated renin levels may be causally associated with the development of LVH in chronic hemodialysis patients. PMID- 9355081 TI - Effect of anti-lymphocyte induction therapy on renal allograft survival: a meta analysis. AB - Induction immunosuppression with antilymphocyte antibodies has not been shown to improve cadaveric kidney allograft survival in randomized, controlled trials despite widespread use. This meta-analysis of randomized, controlled trials assessed the effectiveness of induction therapy in prolonging allograft survival. Studies of induction therapy were identified in Medline (1986 through 1996), using the terms "monoclonal antibodies" or "antilymphocyte serum," and "kidney transplantation," "human," and "clinical trial." Bibliographies, pharmaceutical manufacturers, the United Network for Organ Sharing, National Institutes of Health, and study authors were also consulted. Seven of 247 identified studies met the following inclusion criteria: (1) an adult study population; (2) assessment of antilymphocyte antibodies in the immediate posttransplant period; (3) a control arm of cyclosporine, azathioprine, and prednisone in the immediate posttransplant period; and (4) presentation of survival data. Two readers independently extracted protocol and survival data from each study. Summary odds ratios (fixed and random effects) and a rate ratio from proportional hazards regression at 2 yr were estimated to examine the effect of induction therapy on allograft survival. The summary odds ratios were both 0.66 (confidence interval [CI], 0.45 to 0.96; P = 0.03), and the rate ratio was 0.69 (CI, 0.49 to 0.97; P = 0.03), indicating a beneficial effect of induction therapy on allograft survival. Allograft survival was 85.6% (CI, 82.1 to 89.1%) in the induction therapy group and 79.6% (CI, 75.6 to 83.6%) in the conventional therapy group. These results were stable in a sensitivity analysis based on study quality. Allograft survival was prolonged with induction therapy compared with conventional immunosuppression. These data indicate a potential role for the routine use of induction therapy in renal transplantation to optimize the survival of cadaveric allografts. PMID- 9355082 TI - Aspiration biopsy of the kidney. 1951. PMID- 9355083 TI - Renal insufficiency after intravenous immune globulin therapy: a report of two cases and an analysis of the literature. AB - Over the past decade, intravenous immune globulin therapy (IVIG) has gained widespread use for a variety of clinical disorders. IVIG treatment is associated with a number of complications, including acute renal failure (ARF). Although the cause of IVIG-associated ARF is unknown, it may be related to the stabilizing agent used in the IVIG preparation. The development and resolution of ARF is typically rapid, but is some cases recovery may be delayed and require renal replacement therapy. In such patients, recurrence of ARF may be avoided by selection of a preparation with a different stabilizing agent. Two cases of IVIG induced ARF are described, and all reported cases are analyzed to assess the probable mechanism of renal injury. PMID- 9355084 TI - Goodpasture syndrome involving overlap with Wegener's granulomatosis and anti glomerular basement membrane disease. AB - A 68-year-old Caucasian woman presented to the hospital with nodular pulmonary infiltrates and acute renal failure. Wegener's granulomatosis was initially considered to be most likely because of the presence of increased serum levels of c-anti-neutrophil cytoplasmic antibodies (c-ANCA). A consultation through the Internet after a renal biopsy demonstrated crescentic, necrotizing glomerulonephritis and linear deposits of immunoglobulin G (IgG) and complement C3, typical of anti-glomerular basement membrane (GBM) disease. Hemodialysis was instituted; however, the patient suddenly developed a massive cerebral hemorrhage and died before full therapy could take effect. Postmortem analysis of the patient's sera revealed high titers of IgG against the alpha 3 NC1 domain of type IV collagen. Serologic evidence of both p-ANCA and anti-GBM antibodies are becoming more frequently recognized in the setting of rapidly progressive glomerulonephritis. The patient reported here had the unusual combination of c ANCA antibodies with anti-GBM disease, and this association raises complex questions regarding the pathogenesis of this type of renal injury. PMID- 9355085 TI - Does cytomegalovirus cause glomerular injury in renal allograft recipients? AB - The case of a 16-yr-old woman who received an ABO-incompatible renal allograft for end-stage renal disease due to membranoproliferative glomerulonephritis type I is presented. The patient's posttransplant course was complicated by cytomegalovirus (CMV) infection and the rare finding of glomerular CMV inclusions on renal biopsy. This article focuses on the pathology and pathogenesis of glomerular injury associated with CMV infection in renal allograft recipients and also reviews the epidemiology, clinical implications, diagnosis and management of CMV infection in these patients. PMID- 9355086 TI - Viral infections of nonhuman primates. AB - Approximately 53,000 serologic tests and viral isolation studies were performed on 1,700 nonhuman primate specimens for evidence of past and/or current viral infection. Information, other than the requested test, generally was not provided with the specimen. This lack of information does not permit any attempt at interpretation of results. Requested testing included a large number of diverse viral agents in approximately 40 primate species. The resulting data are in keeping with those of previous studies and offer an insight into the needs of colony management, as well as some general information on the overall frequency of infection with the indicated viruses. Inasmuch as the results represent testing of single specimens, they are not to be construed as "diagnostic," and simply indicate past infection as represented by the presence of antibody in the test animal. Viral isolation results are listed, and the number of positive results versus the number of animals tested emphasizes the limitations of the procedure. Investigations such as these continue to assist in the maintenance of healthy nonhuman primate colonies. This information also supports continued use of nonhuman primates for research in human viral infections and may be helpful in terms of animal selection for use in xenotransplants. PMID- 9355087 TI - Detection of urogenital mycoplasmal infections in primates by use of polymerase chain reaction. AB - Urogenital mycoplasmal infections could affect use of primates as models for reproductive system studies and could affect reproduction in captive primates, but could be useful as animal models of similar human infections. We conducted a pilot study to assess detection of urogenital mycoplasmal infections in primates by use of polymerase chain reaction (PCR). Healthy animals were anesthetized, and vaginal, cervical, or endometrial and urethral swab specimens were collected from females and males, respectively. Specimens were tested by PCR supplemented with dot blotting and nonradiolabeled oligonucleotide probing for 16S rRNA sequences conserved among mollicutes. Specimens with positive results were tested by species-specific PCRs with primers for 16S rRNA sequences of Ureaplasma urealyticum and Mycoplasma hominis and for adhesin gene sequences of Mycoplasma genitalium. Spiked duplicate reactions were included as internal controls for each reaction. Results for 232 specimens from 166 animals indicate that naturally acquired urogenital infections are readily detected and suggest that urogenital mycoplasmal infections are common in laboratory primates (48/166 [29%] overall). M. hominis and U. urealyticum appeared to be common among the studied primates overall and especially in chimpanzees. Mycoplasmas other than M. genitalium, M. hominis, and U. urealyticum appeared to be at least as common as these three, with specimens from 18 of 48 animals (38%) having positive "generic" PCR results, but no positive results in species-specific PCRs. PMID- 9355088 TI - Five spontaneous deaths associated with Clostridium difficile in a colony of cotton-top tamarins (Saguinus oedipus). AB - Clostridium difficile toxin was detected in the feces of five cotton-top tamarins (Saguinus oedipus) that died spontaneously over a period of 10 weeks. Deaths occurred subsequent to antibiotic therapy for infectious diarrhea associated with Campylobacter spp. Relevant clinical signs of disease prior to death included weight loss, watery diarrhea, hematochezia, weakness, and sudden collapse. On histologic examination of the colon at necropsy, pseudomembranous colitis was evident in two cases, a lesion consistent with C. difficile lesions in humans. This finding prompted submission of feces for C. difficile toxin analysis from these five cases. Four of the tamarins were from a single room, and the fifth was housed nearby. The proximity of the cases raises the possibility of environmental contamination by resistant C. difficile spores or fecal spread of the organism as reported in hospitals, day-care centers, and nurseries. The relative importance of C. difficile and its potential role as an unrecognized cause of enteric disease secondary to antibiotic therapy in nonhuman primates is discussed. PMID- 9355089 TI - Anthelmintic treatment to eradicate cutaneous capillariasis in a colony of South African clawed frogs (Xenopus laevis). AB - The nematode Capillaria xenopodis (Pseudocapillaroides xenopi), a skin parasite of South African clawed frogs (Xenopus laevis), is quite common in laboratory animal facilities. It causes serious skin changes and may further lead to wasting and death of affected frogs. Various treatment protocols, using the anthelmintics ivermectin and levamisole, were successively tested for practicability of elimination of the parasite from a colony of clawed frogs. Nematodes were reduced below diagnostic levels by various methods of application of ivermectin (orally or by injection into the dorsal lymph sac, twice at intervals of 10 to 14 days). However, nematodes were found again in the treated animals 1 to 3 months later. Treatment by use of ivermectin-medicated tank water is not feasible due to its low water solubility. Elimination of the parasite was reliably achieved by use of levamisole-medicated tank water. Relapses were not seen during the 18-month posttreatment observation period. Levamisole concentration was 12 mg/L of water, with 4.17, 5.00, or 6.25 L of tank water/frog, and 50, 60, or 75 mg of levamisole available/frog, for at least 4 days, with treatments repeated after 10 to 14 days. Results were reproducible in two trials each with five tanks containing, in turn, four or five frogs each. A treatment trial carried out with a group of 20 adult frogs exposed to 12 mg of levamisole/L of tank water, but with only 2.5 L of tank water/frog (i.e., only 30 mg of levamisole available/animal), was not effective in eradicating the parasites. Not only the drug concentration, but also the amount of drug available per animal seems to be of importance. In contrast to thiabendazole, which is often reported in literature as treatment for cutaneous capillariasis, negative side effects were not observed with use of levamisole. PMID- 9355091 TI - Cardiac troponin T is a sensitive, specific biomarker of cardiac injury in laboratory animals. AB - A reliable serum assay that can discriminate between cardiac and skeletal muscle injury is not available for diagnostic use in laboratory animals. We tested and supported the hypotheses that serum cardiac troponin T (cTnT) was widely applicable in laboratory animals as a biomarker of cardiac injury arising from various causes; that it increased in proportion to severity of cardiac injury; and that it was more cardiospecific than creatine kinase (CK) or lactate dehydrogenase (LD) isozyme activities. In canine and rat models of myocardial infarction, cTnT concentration increased 1,000- to 10,000-fold and was highly correlated with infarct size within 3 h of injury. Serum CK and LD isozymes were substantially less effective biomarkers and, in contrast to cTnT, were ineffective markers in the presence of moderate skeletal muscle injury, with resulting serum CK activity > 5,000 U/L. Using these animal models, and mouse and ferret models, we also showed cTnT to be an effective biomarker in doxorubicin cardiotoxicosis, traumatic injury, ischemia, and cardiac puncture. Reference range serum concentrations for all species were at the detection limit of the assay, except those for mice, in which they were slightly increased, possibly because mice were used to generate assay monoclonal antibodies. We conclude that cTnT is a powerful biomarker in laboratory animals for the sensitive and specific detection of cardiac injury arising from various causes. PMID- 9355090 TI - Endothelial-target rickettsial infection. PMID- 9355092 TI - Cryopreservation of murine zygotes for use in testing culture environments. AB - Developing one-cell mouse zygotes are more sensitive to in vitro environmental conditions than are cleavage-stage embryos. However, for convenience and reproducibility, cryopreserved two-cell zygotes are routinely used for such assays. Concern over the possibility of inducing damage by exposing one-cell zygotes to cryoprotective agents and freeze-thaw procedures during syngamy led us to examine one-cell zygotes, with and without visible pronuclei, in an effort to minimize or avoid these effects and obtain the highest possible developmental rate. In vivo fertilized mouse zygotes were collected 21 to 43 h after administration of human chorionic gonadotropin (hCG). Suspensions of zygotes in 2M ethylene glycol were aspirated into 0.25-ml plastic insemination straws and slowly cooled at -0.5 degree C/min to -40 degrees C before being plunged into liquid nitrogen for storage. Zygotes were thawed, rinsed, and placed in culture. Zygotes were examined initially for damage from the freeze-thaw procedure. Daily in vitro development was recorded. In this group of zygotes, no damage was apparent immediately after thawing, and a high degree of development in vitro was observed. Thus, usefulness of a cryopreservation method for one-cell murine zygotes has been confirmed. PMID- 9355093 TI - Housing and exercise of dogs: effects on behavior, immune function, and cortisol concentration. AB - We examined the effect of supplemental exercise (either individually or with a conspecific) on the physical and psychological health of dogs by measuring immune, endocrine, and behavioral responses. Forty purpose-bred adult male beagles were assigned to one of four treatment conditions: exercised individually (EI), exercised with a conspecific (EC), nonexercised (NE), or cage control (CC). Each EI dog was removed from its cage, carried to an empty room, and allowed to exercise alone for 20 min/d 3 days a weeks for 12 weeks. Two EC dogs were allowed to exercise together following a similar schedule. To control for potential handling effects, NE dogs were removed from their cages, carried to the exercise room, but immediately returned to their cages, and CC dogs remained in their cages during scheduled exercise periods. Detailed behavioral observations, humoral immune responses to the antigen keyhole-limpet hemocyanin, peripheral blood mononuclear cell subsets, plasma cortisol concentration, body weight, and total and differential white blood cell (WBC) counts were routinely monitored. Results indicated few significant treatment effects on physiologic or behavioral measures. Specifically, EC dogs had lower percentages of B lymphocytes, and EC and EI dogs barked more than did NE or CC dogs. However, some physiologic and behavioral measures changed as a function of time regardless of treatment condition. Most notably, for all dogs over time, WBC counts, plasma cortisol values, and behavioral measures reflecting inactivity decreased, while measures reflecting high activity and abnormal behaviors increased. We concluded that neither exercise treatment substantially altered the physical health of research dogs, and perhaps more importantly, failed to prevent the development of abnormal behavior. PMID- 9355094 TI - Light contamination during the dark phase in "photoperiodically controlled" animal rooms: effect on tumor growth and metabolism in rats. AB - Enhanced neoplastic growth and metabolism have been reported in animals maintained in a constant light (24L:0D) environment. Results from this laboratory indicate that tumor growth is directly dependent upon increased ambient blood concentrations of arachidonic and linoleic acids, particularly linoleic acid. Tumor linoleic acid utilization and production if its putative mitogenic metabolite, 13-hydroxyoctadecadienoic acid (13-HODE), are suppressed by the circadian neurohormone melatonin, the production of which is itself regulated by light in all mammals. This study was performed to determine whether minimal light contamination (0.2 lux) in an animal room during an otherwise normal dark phase may disrupt normal circadian production of melatonin and affect tumor growth and metabolism. Animals of groups I (12L:12D), II (12L:12-h light-contaminated dark phase), and III (24L:0D) had plasma total fatty acid (TFA), linoleic acid (LA), and melatonin concentrations measured prior to tumor implantation; groups I and II had daily cycles in plasma TFA and LA values, whereas group III had constant values throughout the day. The integrated mean TFA and LA values for the entire day were similar in all groups. Although group-I animals had a normal nocturnal surge of melatonin (127.0 pg/ml) at 2400 h, the nocturnal amplitude was suppressed in group-II animals (16.0 pg/ml); circadian variation in melatonin concentration was not seen in group-III animals (7.4 pg/ml). At 12 weeks of age, rats had the Morris hepatoma 7288CTC implanted as "tissue-isolated" tumors grown subcutaneously. Latency to onset of palpable tumor mass for groups I, II, and III was 11, 9, and 5 days respectively. Tumor growth rates were 0.72 +/- 0.09, 1.30 +/- 0.15, and 1.48 +/- 0.17 g/d (mean +/- SD, n = 6/group) in groups I, II, and III respectively. Arteriovenous difference measurements for TFA and LA across the tumors were 4.22 +/- 0.89 and 0.83 +/- 0.18 (group I), 8.26 +/- 0.66 and 1.64 +/- 0.13 (group II), and 7.10 +/- 0.78 and 1.50 +/- 0.16 (group III)/min/g, and groups II and III were significantly different from group I (P < 0.05). Tumor TFA and LA contents were 14.3 +/- 1.7 and 1.8 +/- 0.3 (group I), 52.9 +/- 5.5 and 7.9 +/- 0.8 (group II), and 106.0 +/- 12.0 and 18.5 +/- 2.4 (group III) micrograms/g and were significantly different from each other (P < 0.001). Production of 13 HODE by the hepatomas in groups I, II, and III was 35.5 +/- 6.3, 109.6 +/- 10.6, and 196.2 +/- 34.9 ng/min/g respectively, values which also were significantly different among groups (P < 0.001). The results indicate that minimal light contamination of only 0.2 lux during an otherwise normal dark phase inhibits host melatonin secretion and increases the rate of tumor growth and lipid uptake and metabolism. These data suggest that great care must be taken to prevent "light leaks" in animal rooms during the dark phase of a diurnal cycle because such contamination may adversely affect the outcome of tumor growth investigations. PMID- 9355095 TI - Prolonged (12 hours) intravenous anesthesia in the rat. AB - A method is described for prolonging anesthesia in the rat for periods of up to 12 h, with subsequent recovery. Ketamine and xylazine in a ratio of 30 to 1 were infused intravenously by use of an adjustable syringe pump at rates of 40 to 50 microliters/min (1,000 to 1,250 micrograms/kg of body weight/min for ketamine and 32 to 40 micrograms/ kg/min for xylazine), and the rate of infusion was adjusted to maintain a stable depth of anesthesia. Primary assessment of the depth of anesthesia was provided by observation of the respiratory rate, which was maintained between 80 and 100 breaths/min. Rectal temperature was maintained between 35 and 36 degrees C. Blood pressure, pulse, and blood pH and bicarbonate concentration were stable during the 12-h period. The PCO2 increased slightly to 53 mm Hg; PO2 decreased gradually to 59 mm Hg at the last measurement. Plasma glucose concentration decreased progressively; supplemental glucose was given to maintain plasma concentration > 5 mM. After anesthetic administration was discontinued, the animals recovered promptly. PMID- 9355096 TI - Appearance of morphologically abnormal Sertoli cells in infertile PD male rats during postnatal development. AB - Our previous studies using 12-week-old PD (pd/pd) male rats have indicated high incidence of morphologically abnormal Sertoli cells. Because sterility in these males is attributable to abnormal spermatogenesis, it is suggested that the abnormal Sertoli cells are related to abnormal spermatogenesis. In this study, the testes of prepubertal 7- and 21-day-old male pd/pd rats were examined ultrastructurally for the presence of abnormal Sertoli cells. The pd/+ males served as controls. At 7 days of age, light and dark types of Sertoli cells were found in pd/pd males. However, no anatomic abnormality existed in those cells. Cytoplasmic organelles were underdeveloped. The only difference between the light and dark Sertoli cells was the high electron density in the nucleus and cytoplasm of the dark cells. These features were similar between pd/pd and pd/+ males. At 21 days of age, the dark Sertoli cells had disappeared, and light Sertoli cells and abnormal Sertoli cells were observed in pd/pd males. The light Sertoli cells had well-developed cytoplasmic organelles. On the other hand, abnormal Sertoli cells had a dark appearance, irregularly shaped nuclei, and a large quantity of lipid droplets in the cytoplasm. These findings suggested the presence of functional disorders in lipid metabolism. The light and abnormal Sertoli cells also were found in pd/+ males; however, incidence of the abnormal Sertoli cells was significantly higher in pd/pd males (11.3%), compared with that in pd/+ males (4.9%). Thus, these results indicate that in pd/pd male rats, a number of abnormal Sertoli cells appear by 21 days of age. PMID- 9355097 TI - Hematologic and serum biochemical and electrolyte values in clinically normal domestically bred rhesus monkeys (Macaca mulatta) according to age, sex, and gravidity. AB - Age, sex, and gravidity significantly affect blood values in rhesus monkeys (Macaca mulatta). The objective of the study reported here was to provide clinicians and researchers with complete blood values for clinically normal, domestically reared rhesus monkeys of Indian origin and of different age, sex, and gravidity. Values were obtained from 527 healthy, domestically bred and reared rhesus monkeys at The University of Texas M. D. Anderson's Department of Veterinary Sciences, Bastrop, Texas. Because standardized reference values are vital when dealing with the healthcare of a breeding population of rhesus monkeys, it is imperative to have a comprehensive set of reference serum biochemical and hematologic values based on sex, age, and gravidity to determine the overall health status of an individual monkey or a colony used for breeding or research. Establishing reference values allows accurate interpretation of biochemical and hematologic values for normal, subclinical, or clinically ill animals. PMID- 9355098 TI - Diagnostic exercise: neoplastic mass of the vagina and vulva in a dog. PMID- 9355099 TI - Use of endoscopic and ultrasound techniques in the guinea pig leiomyoma model. PMID- 9355100 TI - Multifocal necrotizing enteritis with hepatic and splenic infarction associated with Clostridium perfringens type A in a guinea pig raised in a conventional environment. PMID- 9355101 TI - Speedy backcrossing through in vitro fertilization, using pre-pubertal superovulation and neonatal death dependent on genetic background in angiotensinogen-deficient mice. PMID- 9355102 TI - F344-rnu/rnu athymic rats: breeding performance and acceptance of subcutaneous and intracranial xenografts at different ages. PMID- 9355103 TI - Mycotic thromboses in sheep implanted with biomedical cardiac devices. PMID- 9355104 TI - Facilitation of feedback inhibition through blockade of glucocorticoid receptors in the hippocampus. AB - In the present study the effects of intracerebroventricular (i.c.v.) and intrahippocampal administration of corticosteroid antagonists on basal hypothalamic-pituitary-adrenal (HPA) activity around the diurnal peak were compared in male Wistar rats. In two separate experiments the glucocorticoid receptor (GR) antagonist RU 38486 and the mineralocorticoid receptor (MR) antagonist RU 28318 were tested. One hour after GR antagonist injection, significant increases in plasma ACTH and corticosterone levels were observed in the i.c.v. treated rats, when compared to vehicle. In contrast, a significant decrease in ACTH levels, and a slight, but non-significant decrease in corticosterone concentrations were attained one hour after intrahippocampal injection of the GR antagonist. Injection of the MR antagonist, on the other hand, resulted in enhanced ACTH and corticosterone levels irrespective of the site of injection. These findings suggest that negative feedback inhibition at the circadian peak involves hippocampal MRs and extrahippocampal (hypothalamic) GRs. The latter feedback inhibition overrides a positive feedback influence exerted by endogenous corticosteroids through hippocampal GRs. PMID- 9355105 TI - 5-HT1A receptor agonist reverses adrenalectomy-induced loss of granule neuronal morphology in the rat dentate gyrus. AB - Adrenal steroids are important for maintaining neuronal maturation in the adult rats. Two weeks after bilateral adrenalectomy (ADX), hippocampal MAP-2 (microtubule associated protein-2) and calbindin immunoreactivity (IR) decreased in the molecular layer of the superior blade of the dentate gyrus. The molecular and granular cell layer at the lateral tip of the superior blade decreased in width by 32% and 50%, respectively. The granule neurons showed reduced staining with Nissl and an anti-calbindin antibody. These changes suggested a loss of the mature neuronal morphology. In this same localized regions, two glial proteins, glial fibrillary acidic protein (GFAP) and S-100 beta showed dramatically reduced immunoreactivity. These effects induced by ADX were reduced within 72 hrs by ipsapirone (1 mg/kg), a 5HT1A receptor agonist. Loss of adult neuronal morphology by ADX, and reversal by the 5HT1A agonist, may be evidence of the trophic importance of the 5HT1A receptor in granule neurons of hippocampus. PMID- 9355106 TI - 17 beta-Estradiol enhances the outgrowth and survival of neocortical neurons in culture. AB - Results of this investigation demonstrate that exposure to 17 beta-estradiol differentially and significantly regulates cortical nerve cell outgrowth depending on the cortical region. Parietal and occipital neurons treated with 1 nM 17 beta-estradiol showed a greater magnitude of neuronal outgrowth whereas outgrowth of temporal cortex neurons was decreased in the presence of 1 nM 17 beta-estradiol. Frontal cortex neurons showed a consistent enhancement of neuronal outgrowth that did not reach statistical significance. The dose response profile for 17 beta-estradiol regulation of the macromorphological features exhibited a bimodal dose response relationship whereas the dose response profile for 17 beta-estradiol regulation of the micromorphological features exhibited a dose response more characteristic of an inverted V-shaped function. An antagonist to the NMDA receptor antagonist, AP5, abolished the growth promoting effect of 17 beta-estradiol whereas the nuclear estrogen receptor antagonist ICI 182,780 did not. Lastly, neocortical neurons exposed to 17 beta-estradiol exhibited greater viability and survival than control neurons over a two week period. These data indicate that 17 beta-estradiol can enhance the growth and viability of select populations of neocortical neurons and that the growth promoting effects of 17 beta-estradiol can be blocked by an antagonist to the NMDA glutamate receptor and not by an antagonist to the estrogen nuclear receptor. PMID- 9355107 TI - Expression of the low-affinity p75 nerve growth factor receptor in the developing rat pituitary gland. AB - We investigated, by means of in situ hybridization with a digoxigenin-labelled RNA probe, the expression of the low-affinity p75 nerve growth factor receptor (NGFR) in the developing pituitary primordium of the rat. In 13-day pc embryos, intense staining of p75 NGFR mRNA was present in the cytoplasm of all cells of Rathke's pouch. In day-17 pc embryos p75 NGFR expression was present primarily in the cells of the intermediate lobe. In the newborn rat pituitary only very weak staining was observed, predominantly in the intermediate lobe. In neural structures the staining at day 13 pc was comparable to that of day 17 pc. Since p75 expression is seen very early during pituitary development and declines during the time the expression of pituitary hormonal phenotypes are steadily increasing, we suggest that the p75 NGFR expression in Rathke's pouch may play a temporally defined role in the commitment rather than in the differentiation of the various pituitary cell types. PMID- 9355108 TI - Alterations in glial fibrillary acidic protein (GFAP) mRNA levels in the hamster facial motor nucleus: effects of axotomy and testosterone. AB - Testosterone propionate (TP) administered at the time of facial nerve injury in the hamster accelerates the rate of regeneration. In this study, we tested the hypothesis that the mechanism by which TP augments peripheral nerve regeneration involves regulation of glial fibrillary acidic protein (GFAP) mRNA in the facial motor nucleus. Castrated male hamsters were subjected to right facial nerve transection, with half the animals implanted subcutaneously with Silastic capsules containing exogenous TP and the remainder sham implanted. Postoperative survival times were 0.25, 1, 2, 4, 7, and 14 d. Qualitative/quantitative analyses of both film and emulsion autoradiograms were accomplished. Axotomy, with or without TP, resulted in a dramatic increase in GFAP mRNA levels by 1 d post operative on the axotomized side, relative to controls. GFAP mRNA levels remained elevated throughout all postoperative times in both the nonhormone- and TP treated animals. Qualitative examination of the film autoradiograms indicated a generalized decrease in the amount of GFAP mRNA in the control and axotomized nuclei of TP-treated animals when compared to the control and axotomized nuclei, respectively, of nonhormone-treated animals. Statistical comparison of the values obtained for both the film and emulsion autoradiograms confirmed this impression. Thus, while the injury-induced increases in GFAP mRNA expression were not blocked by TP, the overall extent of the increase was significantly tempered by steroid treatment. These data suggest that hormonal modulation of the astrocytic response to peripheral nerve injury may be a contributing factor in the ability of steroids to enhance the regenerative capacities of injured motor neurons. PMID- 9355109 TI - Gonadal steroid regulation of growth-associated protein GAP-43 mRNA expression in axotomized hamster facial motor neurons. AB - Treatment with testosterone propionate (TP) after nerve injury is known to accelerate both the rate of axonal regeneration and functional recovery from facial paralysis in the adult male hamster. Peripheral nerve injury is also known to increase the expression of a 43 kilodalton growth-associated protein (GAP-43). In the intact brain, GAP-43 expression is affected by gonadal steroids. We thus postulated that steroidal modulation of GAP-43 gene expression may be a component of the neurotrophic action of TP in regenerating neurons. This issue was examined in hamster facial motor neurons (FMN) which contain androgen receptors and which have been shown to respond to exogenous steroids in a number of previous studies. Castrated adult male hamsters were subjected to right facial nerve transection and treated with either TP via subcutaneous hormone capsule implants, or left untreated (no hormone replacement). At post-injury/treatment times of 0.25, 2, 4, 7, and 14 d, the brain stem regions were harvested, cryostat sections were collected through the facial motor nucleus, and in situ hybridization was done using a 33P-labeled GAP-43 cDNA probe. Quantitative analysis of the autoradiograms by computer assisted grain counting revealed that axotomy produced a dramatic increase in GAP-43 mRNA levels in FMN by 2 d post-axotomy and that this increase remained through 14 d post-injury in both the TP-treated and the untreated group. In the nonhormone-treated group, there was a statistically significant dip in GAP-43 mRNA levels in FMN at 7 d post-operative, relative to 4 d post-operative levels. TP-treatment prevented this transient decline in GAP-43 mRNA levels in axotomized FMN. PMID- 9355110 TI - Influence of gonadal steroids on brain corticosteroid receptors: a minireview. AB - Sex differences exist in the functioning of the two brain corticosteroid receptor systems. Ovarian steroid replacement alters receptor mRNA expression, receptor binding capacities, and receptor affinity. The abundance of both mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) message can be reduced by estrogen. Progesterone is able to partially antagonize the action of estrogen and to induce MR transcription. The effect of estrogen on receptor binding capacity is more modest than its transcriptional actions. Estrogen decreases MR binding more reliably than it does GR. Progesterone has high affinity for the MR and can substantially reduce MR affinity for corticoids. Androgen apparently regulates corticoid receptor transcription but may not affect binding capacity. Estrogen and androgen are both more potent in regulating pituitary-adrenal function than would be suggested by their actions on receptor binding parameters. PMID- 9355114 TI - HIV prevention and control in rural areas. PMID- 9355111 TI - The effects of stress on central dopaminergic neurons: possible clinical implications. AB - The response of the central nervous system to stress is often critical to the adaptation of an organism to its environment. However, in humans the response to stress also can be maladaptive, resulting in the expression or exacerbation of many neurological and psychiatric disorders. In this review, we examine the impact of stress on the synthesis and release of dopamine within mesocortical, mesoaccumbens, and nigrostriatal dopamine projections. We note that whereas stress increases the neurochemical activity of each of these populations of dopamine neurons, heterogeneities do exist. Specifically, acute stress evokes a greater increase in dopamine metabolism and release within the prefrontal cortex than the subcortical sites. Furthermore, whereas prior exposure to chronic stress enhances the response of mesocortical dopamine neurons to an acute novel stressor, this does not occur in the subcortical sites. In addition to these regional heterogeneities, we also note that even within a single dopamine projection there can be heterogeneous regulation of dopamine synthesis and release. Specifically, whereas stress-induced dopamine release in the neostriatum is mediated by an action of glutamate on the dopamine cell body, stress-induced dopamine synthesis in the neostriatum is mediated by an action of glutamate on the dopamine nerve terminal. Finally, we propose that regional heterogeneities in the responsiveness of central dopamine neurons to stress may ultimately play a role in the expression and exacerbation of symptoms associated with schizophrenia. PMID- 9355113 TI - Sexually transmitted disease screening in women. PMID- 9355115 TI - Rosacea: recognition and management for the primary care provider. AB - Rosacea is a common facial dermatitis that currently affects an estimated 13 million Americans. It is a chronic and progressive cutaneous vascular disorder, primarily involving the malar and nasal areas of the face. Rosacea is characterized by flushing, erythema, papules, pustules, telanglectasia, facial edema, ocular lesions, and, in its most advanced and severe form, rhinophyma. Ocular lesions are common, including mild conjunctivitis, burning, and grittiness. Blepharitis, the most common ocular manifestation, is a nonulcerative condition of the lid margins. Rosacea most commonly occurs between the ages of 30 to 60, and may be seen in women experiencing hormonal changes associated with menopause. Women are more frequently affected than men; the most severe cases, however, are seen in men. Fair complexioned individuals of Northern European descent are most likely to be at risk for rosacea; most appear to be pre-disposed to flushing and blushing. Alcohol, stress, spicy foods, and extremes of temperature have all been implicated, but have not been found to actually cause rosacea. Early diagnosis by the primary care practitioner, management with systemic antibiotics such as tetracycline, and topical agents such as metronidazole, in conjunction with patient education and lifestyle modifications, can achieve remission in most instances. PMID- 9355116 TI - Endometriosis: pathophysiology, diagnosis, and treatment. AB - Endometriosis is the presence of endometrial tissue outside of the uterine cavity, most commonly surrounding the ovaries and fallopian tubes. It is relatively common disorder in reproductive-age women and is associated with significant pain and morbidity. In most cases, the spread of extrauterine endometrial tissue appears to result from retrograde menstruation and capillary or lymph dissemination. Endometrial cells implanted ectopically respond to cyclical changes in estrogen and progesterone with proliferation and secretion. Their presence in extrauterine areas can initiate immune and inflammatory responses that lead to pain and peritoneal adhesions, and may interfere with fertility. Diagnosis is based on the occurrence of cyclical symptoms and surgical validation via laparoscopy or laparotomy. Treatment is aimed at alleviating pain and preventing complications. Most treatments work by reducing estrogen levels and/or menstrual cycling. A primary practitioner must understand not only the medical aspects of this disease, but the enormous social and psychologic costs as well. PMID- 9355117 TI - Alzheimer's disease: preventing and recognizing a misdiagnosis. AB - The incidence of Alzheimer's disease (AD), representing about 50% of all dementias, is rising in the United States. By the year 2040 there will be an estimated 9 million persons with AD in the United States. The diagnosis of AD is sometimes made prematurely and incorrectly. A significant number of conditions, ranging from the irreversible to the fully reversible, can produce cognitive impairment, and thus may be mistaken for AD. These conditions include other forms of dementia, delirium, and pseudodementia and other psychiatric conditions. Knowledge of the most common of these conditions and their presenting symptoms and precipitating factors, combined with a thorough assessment, can prevent a misdiagnosis of AD. This article reviews several conditions that may be mistaken for AD. Actual case studies (with fictitious names) depict the typical presentation of some of them. PMID- 9355112 TI - Sex and the developing brain: suppression of neuronal estrogen sensitivity by developmental androgen exposure. AB - The developmental effects of androgen play a central role in sexual differentiation of the mammalian central nervous system. The cellular mechanisms responsible for mediating these effects remain incompletely understood. A considerable amount of evidence has accumulated indicating that one of the earliest detectable events in the mechanism of sexual differentiation is a selective and permanent reduction in estrogen receptor concentrations in specific regions of the brain. Using quantitative autoradiographic methods, it has been possible to precisely map the regional distribution of estrogen receptors in the brains of male and female rats, as well as to study the development of sexual dimorphisms in receptor distribution. Despite previous data suggesting that the left and right sides of the brain may be differentially responsive to early androgen exposure, there is no significant right-left asymmetry in estrogen receptor distribution, in either sex. Significant sex differences in receptor density are, however, observed in several regions of the preoptic area, the bed nucleus of the stria terminalis and the ventromedial nucleus of the hypothalamus, particularly in its most rostral and caudal aspects. In the periventricular preoptic area of the female, highest estrogen receptor density occurs in the anteroventral periventricular region: binding in this region is reduced by approximately 50% in the male, as compared to the female. These data are consistent with the hypothesis that androgen-induced defeminization of feminine behavioral and neuroendocrine responses to estrogen may involve selective reductions in the estrogen sensitivity of critical components of the neural circuitry regulating these responses, mediated in part through a reduction in estrogen receptor biosynthesis. PMID- 9355118 TI - Promoting healthy sexuality: guidelines for infancy through preschool. AB - Family is the most important first source of learning about sexuality issues. Parental attitudes and behaviors begin to shape feelings about "maleness" and "femaleness"; thus parents are the focus of teaching healthy sexuality to infants and children. The purpose of this article is to provide the clinician with practical information to promote healthy sexuality in children from infancy through preschool ages. A set of guidelines is presented for the infant, toddler, and preschooler that covers the assessment, rationale, and plan for the clinician. A parental patient education sheet is also presented for each of the age groups (i.e., the infant, toddler, and preschooler). The parental guidelines offer important information for the parent in how to best deal with sexuality issues during each main developmental task. Additionally, a sexuality-values scale is provided. This scale can be used to provide the clinician with information regarding the parents' sexuality knowledge, values, and beliefs (as influenced by their social circumstances, culture, and religion). PMID- 9355119 TI - Clinical implications of alcoholism in patients with HIV/AIDS. PMID- 9355120 TI - The surface glycoconjugates of trypanosomatid parasites. AB - Insect-transmitted protozoan parasites of the order Kinetoplastida, suborder Trypanosomatina, include Trypanosoma brucei (aetiological agent of African sleeping sickness), Trypanosoma cruzi (aetiological agent of Chagas' disease in South and Central America) and Leishmania spp. (aetiological agents of a variety of diseases throughout the tropics and sub-tropics). The structures of the most abundant cell-surface molecules of these organisms is reviewed and correlated with the different modes of parasitism of the three groups of parasites. The major surface molecules are all glycosylphosphatidylinositol (GPI)-anchored glycoproteins, such as the variant surface glycoproteins of T. brucei and the surface mucins of T. cruzi, or complex glycophospholipids, such as the lipophosphoglycans and glycoinositolphospholipids of the leishmanias. Significantly, all of the aforementioned structures share a motif of Man alpha 1 4GlcN alpha 1-6-myo-inositol-1-HPO4-lipid and can therefore be considered to be members of a GPI superfamily. PMID- 9355121 TI - Why intracellular parasitism need not be a degrading experience for Mycobacterium. AB - The success of mycobacteria as pathogens hinges on their ability to infect and persist within the macrophages of their host. However, activation of host macrophages by cytokines from a productive cellular immune response can stimulate the cells to kill their resident pathogens. This suggests that the interaction between host cell and microbe is in delicate balance, which can be tipped in favour of either organism. Biochemical analysis of mycobacterial vacuoles has shown them to be integral to the host cell's recycling endosomal system. As such they show limited acidification and hydrolytic activity despite possession of known lysosomal constituents such as cathepsins D, B and L, and LAMP 1. Even in established infections, they remain dynamic compartments accessible to several plasmalemma-derived constituents. Once the macrophage has been activated by IFN gamma and TNF-alpha the vacuoles coalesce and acidify. This marks a distinct alteration in vacuole physiology and leads to stasis and death of the mycobacteria. Mycobacteria have developed several strategies to avoid this outcome. Most notably, live bacilli-induce sustained release of IL-6 from infected macrophages. IL-6 blocks the ability of both polyclonal primary T cells and T-cell hybridomas to respond to appropriate stimuli. Such an activity could render the centres of infection foci, such as granulomas, anergic and thus avoid release of macrophage-activating cytokines. This paper discusses both the mechanisms by which mycobacteria try to ensure their success as intracellular pathogens and the relevance of these strategies to the overall understanding of mycobacterial diseases. PMID- 9355122 TI - Cytokines and nitric oxide as effector molecules against parasitic infections. AB - Nitric oxide (NO) derived from L-arginine by the catalytic action of inducible NO synthase (iNOS) plays an important role in killing parasites. Many cell types express high levels of iNOS when activated by a number of immunological stimuli which include interferon-gamma (IFN-gamma), tumour necrosis factor alpha, and lipopolysaccharide. IFN-gamma is typically produced by the Th1 subject of CD4+ T cells, whose differentiation depends on interleukin-12 (IL-12) produced by macrophages. Mice with a disrupted iNOS gene were highly susceptible to Leishmania major infection compared with similarly infected control wild-type mice. The mutant mice developed significantly higher levels of TH1-cell response compared with the control mice, suggesting that NO is likely to be the effector molecule in the immunological control of this and other intracellular parasitic infections. To ensure their survival, the Leishmania parasites have evolved effective means to inhibit NO synthesis. The highly conserved major surface glycolipids, glycoinositol-phospholipids and lipophosphoglycan (LPG), of Leishmania are potent inhibitors of NO synthesis. Furthermore, LPG can also inhibit IL-12 synthesis, thereby indirectly blocking the induction of iNOS. The evolutionary and therapeutic implications of these findings are discussed. PMID- 9355123 TI - Genetic analysis of host-parasite coevolution in human malaria. AB - Recent twin studies of clinical malaria and immune responses to malaria antigens have underscored the importance of both major histocompatability complex (MHC) and non-MHC genes in determining variable susceptibility and immune responsiveness. By using a combination of whole genome genetic linkage studies of families and candidate genes analysis, non-MHC genes are being mapped and identified. Human leucocyte antigen (HLA) genotype was found to affect susceptibility to severe malaria in a large study of West African children. T lymphocytes that may mediate such resistance have been identified and their target antigens and epitopes characterized. Some of these epitopes show substantial polymorphism, which appears to result from immune selection pressure. Natural variant epitopes have been found to escape T-cell recognition in cytolytic and other T-cell assays. More recently a novel immune escape mechanism has been described in viral infections, altered peptide ligand antagonism, whereby variants of a T-cell epitope can downregulate or ablate a T cell response to the index peptide. The likely implications of such immune escape mechanisms for the population structure of malaria parasites, for HLA associations with malaria infection and disease, and for the design of new malaria vaccines, are discussed. The evolutionary consequences of such molecular interactions can be assessed by using mathematical models that capture the dynamic of variable host and parasite molecules. Combined genetic, immunological and mathematical analysis of host and parasite variants in natural populations can identify some mechanisms driving host-parasite coevolution. PMID- 9355124 TI - The importance of the back-signal from T cells into antigen-presenting cells in determining susceptibility to parasites. AB - It has long been known that certain MHC class II genes can dominantly suppress immune responses and so increase susceptibility to parasite infections, but the mechanism has been unclear. Recent work has revealed one way in which this form of suppression may operate, through gating by MHC class II molecules of the back signal from activated T cells into macrophages. The two known suppressive genes of the mouse are expressed in macrophages more extensively than are other class II genes. This is associated with suppression of IL-4 production resulting, we infer, from overproduction in the macrophages of IL-12, the counter-cytokine to IL-4. The lack of IL-4 may itself be immunosuppressive, even for Th2 responses, and excess IL-12 can overinduce the antiproliferative cytokine IFN-gamma. Although this mechanism requires further substantiation, we believe that it offers a reasonable answer to an old conundrum. PMID- 9355126 TI - Genetic and biochemical analysis of development in Toxoplasma gondii. AB - Toxoplasma gondii has recently come under intense study as a model for intracellular parasitism because it has a number of properties that facilitate experimental manipulation. Attention is now being turned towards understanding the developmental biology of this complex parasite. The differentiation between the two asexual stages, the rapidly growing tachyzoites and the more slowly dividing, encysted bradyzoites, is of particular interest. Progression from the former to the latter is influenced by the host's immune response. This paper describes current progress on a number of research fronts, all aimed at understanding the triggers that push the tachyzoite-bradyzoite equilibrium in one or other direction and the changes that occur in gene expression (and ultimately metabolism and function). Chief among the techniques used for these studies are genetics and molecular genetics. Recent progress in these areas is described. PMID- 9355125 TI - Immunogenetics of leishmanial and mycobacterial infections: the Belem Family Study. AB - In the 1970s and 1980s, analysis of recombinant inbred, congenic and recombinant haplotype mouse strains permitted us to effectively 'scan' the murine genome for genes controlling resistance and susceptibility to leishmanial infections. Five major regions of the genome were implicated in the control of infections caused by different Leishmania species which, because they show conserved synteny with regions of the human genome, immediately provides candidate gene regions for human disease susceptibility genes. A common intramacrophage niche for leishmanial and mycobacterial pathogens, and a similar spectrum of immune response and disease phenotypes, also led to the prediction that the same genes/candidate gene regions might be responsible for genetic susceptibility to mycobacterial infections such as leprosy and tuberculosis. Indeed, one of the murine genes (Nramp1) was identified for its role in controlling a range of intramacrophage pathogens including leishmania, salmonella and mycobacterium infections. In recent studies, multicase family data on visceral leishmaniasis and the mycobacterial diseases, tuberculosis and leprosy, have been collected from north-eastern Brazil and analysed to determine the role of these candidate genes/regions in determining disease susceptibility. Complex segregation analysis provides evidence for one or two major genes controlling susceptibility to tuberculosis in this population. Family-based linkage analyses (combined segregation and linkage analysis; sib-pair analysis), which have the power to detect linkage between marker loci in candidate gene regions and the putative disease susceptibility genes over 10-20 centimorgans, and transmission disequilibrium testing, which detects allelic associations over 1 centimorgan (ca. 1 megabase), have been used to examine the role of four regions in determining disease susceptibility and/or immune response phenotype. Our results demonstrate: (i) the major histocompatibility complex (MHC: H-2 in mouse, HLA in man: mouse chromosome 17/human 6p; candidates class II and class III including TNF alpha/beta genes) shows both linkage to, and allelic association with, leprosy per se, but is only weakly associated with visceral leishmaniasis and shows neither linkage to nor allelic association with tuberculosis; (ii) no evidence for linkage between NRAMP1, the positionally cloned candidate for the murine macrophage resistance gene Ity/Lsh/Bcg (mouse chromosome 1/human 2q35), and susceptibility to tuberculosis or visceral leishmaniasis could be demonstrated in this Brazilian population; (iii) the region of human chromosome 17q (candidates NOS2A, SCYA2-5) homologous with distal mouse chromosome 11, originally identified as carrying the Scl1 gene controlling healing versus nonhealing responses to Leishmania major, is linked to tuberculosis susceptibility; and (iv) the 'T helper 2' cytokine gene cluster (proximal murine chromosome 11/human 5q; candidates IL4, IL5, IL9, IRF1, CD14) controlling later phases of murine L. major infection, is not linked to human disease susceptibility for any of the three infections, but shows linkage to and highly significant allelic association with ability to mount an immune response to mycobacterial antigens. These studies demonstrate that the 'mouse-to-man' strategy, refined by our knowledge of the human immune response to infection, can lead to the identification of important candidate gene regions in man. PMID- 9355127 TI - Mechanisms of innate resistance to Toxoplasma gondii infection. AB - The interaction of protozoan parasites with innate host defences is critical in determining the character of the subsequent infection. The initial steps in the encounter of Toxoplasma gondii with the vertebrate immune system provide a striking example of this important aspect of the host-parasite relationship. In immuno-competent individuals this intracellular protozoan produces an asymptomatic chronic infection as part of its strategy for transmission. Nevertheless, T. gondii is inherently a highly virulent pathogen. The rapid induction by the parasite of a potent cell-mediated immune response that both limits its growth and drives conversion to a dormant cyst stage explains this apparent paradox. Studies with gene-deficient mice have demonstrated the interleukin-12 (IL-12)-dependent production of interferon gamma (IFN-gamma) to be of paramount importance in controlling early parasite growth. However, this seems to be independent of nitric oxide production as mice deficient in inducible nitric oxide synthase (iNOS) and tumour necrosis factor receptor were able to control early growth of T. gondii, although, they later succumbed to infection. Nitric oxide does, however, seem to be important in controlling persistent infection; treating chronic infection with iNOS metabolic inhibitors results in disease reactivation. Preliminary evidence implicates neutrophils in effector pathways against this parasite distinct from that described for macrophages. Once initiated, IL-12-dependent IFN-gamma production in synergy with other proinflammatory cytokines can positively feed back on itself to induce 'cytokine shock'. Regulatory cytokines, particularly IL-10, are essential to down-regulate inflammation and limit host pathology. PMID- 9355128 TI - Pre-erythrocytic-stage immune effector mechanisms in Plasmodium spp. infections. AB - The potent protective immunity against malaria induced by immunization of mice and humans with radiation-attenuated Plasmodium spp. sporozoites is thought to be mediated primarily by T-cell responses directed against infected hepatocytes. This has led to considerable efforts to develop subunit vaccines that duplicate this protective immunity, but a universally effective vaccine is still not available and in vitro correlates of protective immunity have not been established. Contributing to this delay has been a lack of understanding of the mechanisms responsible for the protection. There are now data indicating that CD8+ T cells, CD4+ T cells, cytokines, and nitric oxide can all mediate the elimination of infected hepatocytes in vitro and in vivo. By dissecting the protection induced by immunization with irradiated sporozoite, DNA and synthetic peptide-adjuvant vaccines, we have demonstrated that different T-cell-dependent immune responses mediate protective immunity in the same inbred strain of mouse, depending on the method of immunization. Furthermore, the mechanism of protection induced by a single method of immunization may vary among different strains of mice. These data have important implications for the development of pre erythrocytic-stage vaccines designed to protect a heterogeneous human population, and of assays that predict protective immunity. PMID- 9355129 TI - Antigenic variation in Giardia lamblia and the host's immune response. AB - Giardia lamblia, a protozoan parasite of the small intestine of humans and other animals, undergoes surface antigenic variation. The antigens involved belong to a family of variant-specific surface proteins (VSPs), which are unique, cysteine rich zinc finger proteins. The patterns of infection in humans and animals fail to show the expected cyclical waves of increasing and decreasing numbers of parasites expressing unique VSPs. Nevertheless, changes in VSP expression occur within the population in vivo owing to selection of VSPs by both immune and non immune mechanisms. After inoculation of a single G. lamblia clone (able to persist in the absence of immune pressure) expressing one VSP (> or = 90%) into mice or humans, the original VSP continues to be expressed until 2 weeks post inoculation (p.i.), when many other VSPs gradually replace it. Selection by immune-mediated processes is suggested because switching occurs at the same time that humoral responses are first detected. In most mouse strains, switching also occurs at about two weeks. Almost all trophozoites are eliminated at three weeks (p.i.), but a barely detectable infection persists over months. In neonatal mice, apparent self-cure is delayed until the sixth or seventh week. Antigenic switching does not occur in adult or neonatal severe combined immunodeficiency disease (SCID) mice, but does occur in neonatal nude mice, thus implicating B cell-mediated mechanisms in immune switching. Not all VSPs are expressed to the same degree in vivo. Some VSPs appear to be preferentially selected whereas others are eliminated on a non-immune basis. In infections in which immunity does not play a role, such as in SCID mice, and during the first week of infection in immunocompetent mice or gerbils, persisting VSPs are preferentially expressed and maintained whereas non-persisting VSPs are replaced within the first week of infection. The purpose of antigenic variation may be presentation of a wide assortment of VSPs to hosts, increasing the chance of a successful initial infection or reinfection. Immune selection of variants comes into play following biological selection. PMID- 9355132 TI - Assessing outcomes in clinical practice. AB - We are witnessing a remarkable explosion in interest and activity in quantifying outcomes and using these measures to enhance the value of clinical care. Outcomes assessment has become an imperative for clinical practice. This paper first will offer criteria for an ideal system of outcomes assessment. The paper will then review the principal domains of assessment for psychiatric practice and provide examples of instruments available in each domain. We will then describe the use of two instruments, one for clinical outcome and one for interpersonal aspects of patient satisfaction, developed and used at McLean Hospital. The relation between outcomes assessment and outcomes management will then be discussed. Finally, we will discuss the fundamental questions a clinical group or facility might consider in choosing outcomes measurement instruments. PMID- 9355130 TI - Th2-mediated host protective immunity to intestinal nematode infections. AB - Despite many years of study, relatively little is known about the effector mechanisms that operate against intestine-dwelling nematodes. Most of the current understanding comes from studies of laboratory model systems in rodents. It is clear that when an intestinal helminth infection takes place the immune system generates a strong Th2-mediated response, which regulates a variety of responses characteristic of helminth infections such as eosinophilia, intestinal mastocytosis and elevated IgE production. The ability to modulate the host's immune response in vivo with cytokine-specific monoclonal antibodies and recombinant cytokines, together with the use of animals with disruption of key genes involved in the immune response, have provided powerful tools with which to dissect the potential effector mechanisms operating. In the absence of a T-cell compartment the host is unable to expel the parasite. If a Th1-dominated response is generated, protective immunity is almost universally compromised. Thus, it it would appear that some aspect of a Th2-mediated response controls effector mechanisms. Although it is clear that for some infections the mast cell appears to be involved in protection, probably through the generation of a non-specific inflammatory response, how these cells become activated remains unclear. Data from infections in transgenic animals suggest that activation is not through the high-affinity receptor for IgE. Such studies also call into doubt the importance of conventional interactions between effector leucocytes and antibody. There is little evidence to support a protective role for eosinophilia in any system. New data also imply that, although interleukin 4 (IL-4) is generally important (and can exert effects independent of an adaptive immune response), it is not always sufficient to mediate protection; other Th2 cytokines (e.g. IL-13) may warrant closer investigation. It is apparent that a number of potential Th2-controlled effector mechanisms (some of which may be particularly important at mucosal surfaces) remain to be explored. Overall, it is likely that worm expulsion is the result of a combination of multiple mechanisms, some of which are more critical to some species of parasite than to others. PMID- 9355133 TI - Violence and psychiatric disorders: results from an epidemiological study of young adults in Israel. AB - This paper investigates the association between various psychiatric disorders and violent behavior using data from a community-based epidemiological study of young adults in Israel (N = 2678). Self-reports of recent fighting and weapon use were elevated among respondents diagnosed with psychotic or bipolar disorders but not among those diagnosed with non-psychotic depression, generalized anxiety disorder or phobias compared to respondents without these disorders. Violence was measured using the Psychiatric Epidemiology Research Interview; psychiatric disorders were diagnosed using a modified version of the Schedule for Affective Disorders and Schizophrenia. The analyses controlled for lifetime substance abuse, antisocial personality disorder and demographic characteristics, thereby extending support for a causal connection between some types of psychiatric disorders and violence. The association between disorder and violence was stronger among respondents with less education, indicating the potentially important role of social and cultural contexts in moderating the association between mental illness and violence. PMID- 9355131 TI - Host-parasite interaction and morbidity in malaria endemic areas. AB - Severe morbidity due to Plasmodium falciparum is a major health problem in African children. The patterns of morbidity in endemic areas are modified by the immune response, and vary markedly with transmission intensity. Severe disease falls into three overlapping syndromes: coma, respiratory distress, and severe anaemia. Recently, it has become clear that metabolic acidosis plays a major role in the pathogenesis of severe disease and is particularly important in the overlap between the different clinical syndromes. We propose that the different manifestations of severe malarial morbidity arise from the interaction of a limited number of pathogenic processes: red cell destruction, toxin-mediated activation of cytokine cascades, and infected cell sequestration in tissue microvascular beds. The pattern of severe morbidity varies with age within any one endemic area, with severe anaemia predominating in the youngest children and coma having its highest incidence in older children. Between endemic areas there is a marked variation in mean age of children with severe malaria, and therefore in the importance of different clinical syndromes. The shift in mean age is due to a combination of increased challenge and more rapid development of immunity at higher levels of transmission. Recent comparative studies indicate that at higher levels of transmission the net effect of these shifts may be a paradoxical reduction in total severe malarial morbidity. PMID- 9355134 TI - Attention and clinical symptoms in schizophrenia. AB - Attentional deficits, long established to characterize patients with schizophrenia spectrum disorders, have traditionally been regarded as part of the disorder's clinical syndrome. In this paper we provide evidence to indicate that: a) impaired attention is a dimension of schizophrenia that is independent of clinical state, and b) that attention does not appear to respond to the medication (i.e. standard neuroleptics) most typically used to treat clinical symptoms. Since intact attention and other cognitive processes appear critical to successful functioning in the community after hospital discharge, these findings have major implications for treatment. PMID- 9355136 TI - Assessment and treatment selection for "revolving door" inpatients with schizophrenia. AB - GOALS: The goals of this study are 1) to determine causes and patterns of relapse for a cohort of "revolving door" schizophrenia inpatients, and 2) to assess the feasibility of starting a new psychopharmacologic intervention before discharge, either depot therapy or an atypical antipsychotic. METHODS: Consecutive admissions to an acute inpatient unit in New York City were screened for "revolving door" criteria. Patients had to have a primary diagnosis of schizophrenia or schizoaffective disorder and have either 1) two hospitalizations in the last year, or 2) three hospitalizations in the last three years. Patients were then assessed for probable causes of relapse for the index and prior two hospitalizations. Treatment selection, based on this information, was trichotomized to: 1) oral conventional antipsychotic, 2) depot conventional antipsychotic (either haloperidol or fluphenazine decanoate), or 3) atypical antipsychotic (either risperidone or clozapine). RESULTS: Sixty-three out of 131 screened admissions met the above revolving door criteria. They were indeed "revolving", having an average of 1.3 hospitalizations per year over the last 3 years and were only out of the hospital for five months (median) before index admission. The treatment selection process was hampered by lack of information about events leading to relapse, and by the lack of outpatient participation in the medication selection process. Of the 50 patients with complete histories about precipitants for the index episode, the most common reason for rehospitalization was judged to be medication noncompliance (n = 25; 50%), followed by medication nonresponse (n = 13; 26%). Not surprisingly, medication recommendations were closely linked to the assessed reason for relapse (depot therapy [n = 27; 49%] with medication noncompliance; atypical antipsychotic [n = 20; 37%] with medication nonresponse [X2 = 26.9, p < .001]). These two recommendations were implemented before discharge for about one-half of the cases. Patient refusal was a relatively greater problem for depot recommendation while constraints in the outpatient environment were more problematic for patients recommended for atypical antipsychotics. CONCLUSIONS: Medication noncompliance and medication nonresponse, in that order, were judged to be the most common causes of relapse for "revolving door" inpatients. Both depot therapy and atypical antipsychotics were commonly recommended and ultimately accepted by about 2/3rds of patients. Choice between depot and atypical was driven by the assessed cause of relapse. In summary, it seems possible to identify "revolving door" inpatients, and to target specific medication interventions within the time frame of an acute inpatient admission. PMID- 9355135 TI - Assessment and intervention for the suicidal patient with schizophrenia. AB - The risk for suicidal behavior in schizophrenia is high with 10-15% committing suicide and 20-40% making suicide attempts. Due to the chronicity and complexity of schizophrenia and the multi-determined nature of suicidal behavior, the clinician must utilize a biopsychosocial approach to assessment and intervention. Clinical factors such as psychosis, depression and substance abuse increase the risk for suicidal behavior in schizophrenia. Social factors such as social adjustment and social supports also play a critical role. Ongoing assessment and intervention of suicidal behavior, clinical symptomatology, social environment and treatment issues are essential. Prediction and prevention of suicidal behavior are not always possible however. Treatment focused on the reduction of symptomatology and maintenance of an effective social environment may attenuate the risk for suicidal behavior in schizophrenia. PMID- 9355137 TI - Psychosocial rehabilitation for the mentally disabled: what have we learned? PMID- 9355138 TI - Effects of short storage conditions and broiler breeder age on hatchability, hatching time, and chick weights. AB - An experiment was conducted to assess how hatching performance is affected by breeder age and egg holding environment during short-term storage. Response variables analyzed were egg weight loss up to 18 d of incubation, viability (hatchability of fertile eggs), embryonic mortality, hatching time, and weight of male and female chicks, at hatching and at the end of incubation. The trials involved a total of 2,250 hatching eggs from each of two commercial broiler breeder flocks of the same strain (Avian) but of different ages (32 to 34 and 48 to 50 wk). Eggs were stored for 0, 1, or 2 d in the egg storage room or in the setter room. The hatching times of the chicks were recorded at 4-h intervals during the period from 478 to 494 h postincubation, and at 514 h, when incubation was terminated and all chicks were removed from the hatcher. In eggs from younger hens, viability was not influenced by preincubation storage; in older hens, viability of eggs not submitted to storage was higher (P < 0.05) by 3 to 6 percentage points than that of stored eggs. Hatching times were not affected by age of the hen, whereas male chicks tended to hatch, on average, about 3 h later than females. Chick weights at hatching and at removal from the hatcher were similar for both sexes, but females experienced a higher (P < 0.05) weight loss in that interval. Eggs incubated on the day of lay tended to hatch, on average, later than stored eggs (especially when compared to eggs submitted to 1 d storage), and produced heavier chicks. PMID- 9355139 TI - Self-selection of dietary protein and energy by broilers grown under a tropical climate: effect of feed particle size on the feed choice. AB - Broilers, 2 wk of age, that had been previously adapted to energy: protein choice feeding, were offered corn (either ground, cracked, or presented as whole grains) and a protein concentrate (43.7% CP) in mash or pellet form. When corn was fed as whole grains, protein concentrate in the selected diet was significantly higher (35.1%) than with cracked corn (29.3%) or ground corn (29.1%). Presenting the concentrate as pellets resulted in a significantly higher concentration in the diet (32.7%) than when mash concentrate was fed (29.6%). Live BW at 4 and 6 wk of age were not significantly affected by feed texture. However, offering corn as whole grains or concentrate as pellets induced a significant improvement in feed efficiency. Total time to eat larger size particles (whole grains, pelleted concentrate) was significantly less than total time to eat ground corn or mash concentrate. Furthermore, the mean duration of the feeding bouts was two times shorter for whole grains (48 s) than for ground corn (98 s) and for pelleted concentrate (56 s) than for mash concentrate (114 s). Chickens ate whole grains or pellets at a significantly slower rate (number of pecks per second feeding time) than when eating ground corn or mash concentrate. There was a rejection during the first 24 h when the form of the concentrate (mash to pellets) was changed. Full adaptation to the new size of the concentrate required about 3 d. PMID- 9355140 TI - Drinking water contaminants (arsenic, cadmium, lead, benzene, and trichloroethylene). 1. Interaction of contaminants with nutritional status on general performance and immune function in broiler chickens. AB - The objective of this study was to examine possible interactions between drinking water contaminants and suboptimal nutritional status for performance and immune function in male broiler chickens. Experimental drinking water contained a mixture of arsenic, benzene, cadmium, lead, and trichloroethylene (TCE) at low concentrations (0.80, 1.3, 5.0, 6.7, and 0.65 ppm) and high concentrations (8.6, 13, 50, 67, and 6.5 ppm). These chemicals were selected because they are among the most common contaminants found in ground water near hazardous waste sites. The experimental diets included feed containing 50% added vitamins and minerals (V&M) and feed without added V&M. Increasing levels of drinking water contaminants and decreasing levels of V&M in diet resulted in significantly (P < or = 0.05) decreased water and feed intake, decreased weight gain, and suppression of natural, humoral, and cell-mediated immune response. In a paired water study, feed consumption, body weight, and immune function were decreased in chickens provided low and high concentrations of the chemical mixture in drinking water compared with chickens given control drinking water equal to the volumes consumed by the chickens given the low and high concentration of mixture, respectively. A deficiency of dietary V&M caused increased sensitivity to adverse effects of drinking water contaminants. PMID- 9355141 TI - Drinking water contaminants (arsenic, cadmium, lead, benzene, and trichloroethylene). 2. Effects on reproductive performance, egg quality, and embryo toxicity in broiler breeders. AB - Broiler breeder hens were used to determine the effect of drinking water containing a low concentration of a chemical mixture (arsenic, 0.8 ppm; benzene, 1.3 ppm; cadmium, 5.1 ppm; lead, 6.7 ppm; and trichloroethylene, 0.65 ppm) and a high (10 times greater than the low concentration of the chemical mixture) levels of the chemical mixture. These chemicals are present in ground water near hazardous waste sites. Water consumption significantly decreased in chickens provided the high concentration of the chemical mixture, whereas feed consumption was not affected in any treatment. There was a linear relationship between increasing concentration of the chemical mixture in drinking water and decreasing body weight of hens. The low concentration of the chemical mixture significantly decreased egg production and egg weight, and increased percentage embryonic mortality. These results suggest that reproductive function in hens is sensitive to adverse effects of contaminated drinking water. PMID- 9355142 TI - The use of competitive exclusion in broilers to reduce the level of Salmonella contamination on the farm and at the processing plant. AB - The effect of a competitive exclusion (CE) product, Broilact, on Salmonella contamination of broiler chickens was studied on the farm and at the processing plant. In the first part of the study, two flocks per week, a CE-treated and an untreated control flock, were placed in similar broiler houses. The CE treatment was administered in the hatchery using a modified spray vaccination cabinet. Salmonella was analyzed from the paper pads of the transport boxes on arrival at the farm and from fecal samples taken 2 wk before slaughter. The results of Salmonella sampling were received for 67 flocks. The other 141 flocks of the company that were reared during the trial period were also sampled for Salmonella and the results were compared to those of treatment and control groups. Broiler performance, including mortality, weight, and feed conversion, was recorded for the trial flocks. In the second part of the study, Salmonella contamination of neck skin samples taken at the processing plant from 18 CE-treated and 28 control flocks was compared. The Broilact-treatment significantly reduced Salmonella contamination both on the farm and at the processing plant. At the level of the farm, the percentage of Salmonella-positive flocks was essentially the same in the control flocks and in other flocks reared during the trial period. An improvement in broiler performance was indicated, although the difference was not significant. PMID- 9355143 TI - Effect of dietary dl-alpha-tocopherol on tissue alpha- and gamma-tocopherol and pulmonary hypertension syndrome (ascites) in broilers. AB - The objectives of this experiment were to determine the effects of high dietary levels of vitamin E on growth performance and pulmonary hypertension syndrome (PHS) mortality. Male broiler chicks (Cobb 500) were randomly assigned to one of four dietary treatments consisting of standard starter and grower diets supplemented with 0, 17, 46, and 87 mg dl-alpha-tocopherol acetate/kg. To encourage the development of PHS, air temperature in the house was 32 and 28 C for Weeks 1 and 2, dropped to 18 C during Week 3, and kept between 10 and 15 C during Weeks 4 through 7. Also, chicks were placed in floor pens on litter used for five previous flocks and ventilation reduced to increase dust and ammonia in the house. Ammonia levels increased from an initial 18 to 36 ppm on Day 42 with the increase in ammonia corresponding to an obvious increase in dust in the air. Lung and liver tissue obtained at 2, 5, and 7 wk of age were analyzed for tissue alpha- and gamma-tocopherol by liquid chromatography. Dietary vitamin E had no effect on body weight, feed intake, or feed efficiency. Cumulative PHS mortality through 7 wk of age was 21% and was also unaffected by dietary treatment. Liver and lung alpha-tocopherol concentrations exhibited a dose-response increase to dietary tocopherol and there was a high correlation between lung and liver tissue alpha-tocopherol (r = 0.72, P < 0.05). Whereas gamma-tocopherol concentrations in lung and liver were unaffected by dietary treatment, liver and lung exhibited age dependent increases in both alpha- and gamma-tocopherol. Despite dose-dependent increases in tissue alpha-tocopherol, supplementation of diets with up to 87 mg dl-alpha-tocopherol acetate had no effect on growth performance or PHS mortality in broilers under the conditions used in this study. PMID- 9355144 TI - Probabilistic neural network prediction of ascites in broilers based on minimally invasive physiological factors. AB - A Probabilistic Neural Network (PNN) was trained to predict ascites in broilers based on minimally invasive inputs (i.e., physiological factors that do not require the death of the bird). A PNN is a supervised, three-layer, artificial neural network that classifies input patterns (e.g., physiological data) into specific output categories (e.g., ascites or no ascites). The PNN inputs were O2 level in the blood, body weight, electrocardiogram (ECG), hematocrit, S wave, and heart rate of individual birds. These data were from three experiments that have been described previously (Roush et al., 1996a,b). The three data sets were pooled into a combined data set for a total of 170 observations. From the pooled data, a training set (117 birds), a calibration set (17 birds), and a verification set (36 birds) were extracted. The PNN was trained on the training data set. To prevent the PNN from overfitting the training data, the neural network was evaluated on its ability to make correct predictions of the calibration data set. At the point at which the neural network made the highest number of correct classifications for the calibration data set, the trained neural network was saved on the computer. When the PNN was applied to the complete data set, the sensitivity or proportion of the birds with ascites that the PNN correctly diagnosed was 0.97 (75/77 birds). The specificity or proportion of birds that the PNN made a correct diagnosis of not having ascites was 0.98 (91/93 birds). When the PNN was applied to the verification data set, which was not subjected to neural network training, the sensitivity was 0.95 (19/20) and the specificity was 0.88 (14/16 birds). Use of models developed with artificial neural networks may enhance the diagnosis of ascites in broilers. The results may be useful in choosing and developing broiler strains that do not have a propensity for ascites. PMID- 9355145 TI - Major histocompatibility complex class II DNA polymorphisms in chicken strains selected for Marek's disease resistance and egg production or for egg production alone. AB - The objective of this study was to investigate frequencies of major histocompatibility complex (MHC) class II restriction fragments in two groups of White Leghorn strains. Each group consisted of an unselected control, a strain selected for egg production traits, and a strain selected for egg production traits and Marek's disease (MD) resistance. PvuII-digested genomic DNA was hybridized with a chicken genomic MHC class II probe. The MHC class II DNA fragment frequencies in the selected strains differed from those in the related unselected control and in the strain selected using the same criteria from a different base population. Based on the sizes of the breeding populations, particularly those in the control strain and in the strain selected for egg production, it was considered unlikely that the observed changes of the MHC class II fragment frequencies were due to random genetic drift. The data suggested that some MHC class II bands are associated with production traits or with MD resistance, and that these associations tend to be unique to each genetic background. Hence, MHC class II genes are likely candidates for the investigation of quantitative trait loci in egg production and disease resistance traits such as those for which the studied strains were selected. PMID- 9355146 TI - Nitric oxide synthase activity and mRNA expression in chicken macrophages. AB - The activity of inducible nitric oxide synthase (iNOS) enzyme was quantified in chicken macrophages. Macrophages from Cornell K-strain (B15B15), GB1 (B13B13), and GB2 (B6B6) chickens and a transformed cell line (MQ-NCSU) were incubated with or without varying concentrations of bacterial lipopolysaccharide (LPS). The culture supernatants were tested for the presence of nitrite. Macrophages from either source produced minimal nitrite (< 4.4 microM/1 x 10(6) cells) levels without LPS stimulation. However, nitrite levels produced by K-strain (42 microM) and MQ-NCSU (41 microM) macrophages were higher (P < 0.05) than those produced by the GB1 (14 microM) and GB2 (14 microM) per 1 x 10(6) macrophages with optimum LPS concentration range of 50 ng to 1 microgram/mL. The addition of an L-arginine analog, NGMMLA, at a concentration of 200 microM completely abolished nitrite production. The addition of 10% vol/vol lymphokines exhibited an additive effect on nitrite production in conjunction with LPS. The increased nitrite production by the K-strain and MQ-NCSU macrophages corresponded to an increased expression of iNOS mRNA as compared to the mRNA produced by GB1 and GB2 macrophages. The iNOS mRNA kinetics study revealed that mRNA levels peaked between 6 to 12 h. The cells from avian lymphoid lineage failed to produce any detectable iNOS activity. These studies showed that macrophages from varying sources differ in NOS activity and implied that genetic background may dictate the extent of arginine-mediated contribution in various biological and immunological functions. PMID- 9355147 TI - Effect of dried solids of nejayote on broiler growth. AB - The purpose of the present study was to test the suitability of the solids of nejayote (a waste product from the tortilla industry) in diets for broilers. The nejayote was obtained from two different tortilla-making factories and the solids were obtained by centrifuge then dried in a hot-air drier. Diets were formulated to be isocaloric and isonitrogenous according to the NRC dietary requirements (1994). Nejayote solids were supplemented at 2, 4, and 6% of the diet. Results show that the content of protein and calcium in the dried solids of nejayote were 5 and 13%, respectively. The performance of broilers fed diets supplemented with dried nejayote did not differ from that of those fed the control diet. Therefore, it is concluded that nejayote solids are suitable for broiler feed and do not affect growth performance. Utilization of nejayote solids at higher levels is a possibility provided that no adverse effects on body weight, feed utilization, and feed:gain ratios are observed. PMID- 9355148 TI - The efficacy of phytase in corn-soybean meal-based diets for laying hens. AB - Microbial phytase hydrolyzes poorly degradable vegetable phytate P in the gastrointestinal tract of poultry; thereby increasing the availability of organic P to an extent that remains to be established. For this purpose, the P equivalency value of phytase in corn-soybean meal layer diets was assessed in three experiments (two short-term absorption studies and one performance trial lasting a complete production period). In the first absorption study, two basal diets containing 30 or 40 g Ca/kg diet were supplemented with either phytase [0, 250, or 500 phytase units (FTU)/kg diet] or with monocalcium phosphate (MCP; 0, 0.5, or 1.0 g P/kg diet) and fed to layers from 20 to 24 wk of age. The ileal absorption of Ca and P was measured during the last week. It was shown that 250 FTU/kg diet hydrolyzed an amount of phytate P that was equivalent to 1.3 g P from MCP. At the highest phytase inclusion level (500 FTU/ kg diet), a lower value of equivalency was observed, as P absorption was almost maximal at the lower level of phytase inclusion (250 FTU/kg diet). Phytase hydrolyzed phytate-bound P effectively at both Ca levels, although this degradation was significantly reduced by 12 percentage units at the higher dietary Ca level. The second absorption study, used 0, 250, and 500 FTU phytase/kg diet and 0 and 1.0 g P/kg diet of MCP. All diets were standardized at 35 g Ca/kg diet. The ileal absorption of Ca and P was determined at 24 and 36 wk of age. These values were significantly reduced in 36-wk-old hens compared to 24-wk-old hens. At 24 wk of age, phytic acid P degradation was significantly improved with increasing levels of phytase up to the maximum inclusion level of 500 FTU/kg diet (maximum phytic acid-P degradation at the end of the small intestine was 66%). In this experiment, the dose of 250 FTU/kg diet was equivalent to 0.8 g MCP-P. In Experiment 3, either phytase or MCP-P was added to a corn-soybean meal layer diet, containing 40 g Ca/kg diet and 3.6 g P/kg diet, at levels of 0, 100, 200, and 300 FTU/kg or levels of 0, 0.3, 0.6, and 0.9 g MCP-P/kg, respectively. Production performance was measured from 18 to 68 wk of age. Diets were consumed ad libitum. Growth, production performances (except kilograms of feed per kilogram of egg), and tibia parameters were significantly improved by dietary supplementation of the negative control diet with either phytase or MCP-P. Growth, egg production, and feed conversion ratio of the hens from the supplemented groups remained good throughout the experiment. No phytase dose effects on the production characteristics or tibia parameters were observed, indicating that the P requirements of the laying hens were met throughout the production period even at the lowest level of supplementation. PMID- 9355149 TI - Response of two strains of large white male turkeys to amino acid levels when diets are changed at three- or four-week intervals. AB - A study was conducted to evaluate the amino acid recommendations of the NRC (1994) when diets were changed at 3- or 4-wk intervals. Diets formulated to provide from 90 to 115% of recommended amino acid levels were fed to Nicholas (NIC) and British United Turkey (BUT) Large White males from day-old to 24 wk of age; samples of turkeys were processed at 18 and 24 wk. Results indicated that the amino acid levels suggested by NRC are adequate to support maximum body weight gain, feed conversion, and dressing percentage of Large White males grown to 18 or 24 wk when fed on 4-wk intervals. An approximately 5% higher level of amino acids was required to maximize breast yield. When feeds were changed at 3 wk intervals, higher levels of amino acids were required to maximize performance; however, there did not seem to be any difference in the response of the two strains of turkeys to different levels of amino acids in this study. PMID- 9355150 TI - Effects of an oxygen-enriched environment on the survival of turkey embryos between twenty-five and twenty-eight days of age. AB - The hypothesis was tested that increased partial pressure of oxygen during the plateau (25 to 26 d of incubation for turkeys) and paranatal (27 to 28 d of incubation) stages of incubation may increase survival rates of turkeys from selected genetic lines. Partial pressure of oxygen inside the incubator cabinet was increased to 171 + 3 mm Hg of the barometric pressure during the plateau stage in oxygen consumption and compared to ambient oxygen (152 + 3 mm Hg). Turkey embryos from genetic lines selected for egg production (E) or growth (F) were compared to their respective randombred controls. These genetic lines have previously been shown to differ in egg weight, eggshell conductance, length of incubation period, embryonic gluconeogenesis, and survival rates during late incubation. Blood, liver, heart, and pipping muscle samples were obtained prior to pipping, at internal pipping and external pipping, and at hatching. The blood was analyzed for glucose concentration and the remaining tissues were assayed for glycogen concentrations. Survival rates were determined on approximately 2,200 eggs in each of three independent trials of the experiment. Interactions of oxygen treatment and genetic line were observed for embryonic survival, heart growth, and hepatic glycogen content. The data suggest that the response to increased oxygen tension in selected genetic lines has been diminished. It was concluded that embryos have been altered metabolically by genetic selection and the concomitant increase in mortality of selected lines during the plateau and paranatal stages is not simply the result of shell quality and hypoxia. PMID- 9355152 TI - Changes in the population of proliferating cells in chicken anterior pituitary during induced molting: an immunocytochemical analysis for proliferating cell nuclear antigen. AB - The goal of this study was to determine whether cell proliferation is increased in the anterior pituitary during an induced molting period in chickens. The anterior pituitaries were collected from: 1) laying hens before initiation of an induced molt (control hens), 2) molting hens 2 to 16 d after cessation of lay, and 3) second cycle laying hens 1 d after returning to lay. Pituitary cephalic and caudal lobes were processed for immunocytochemistry of proliferating cell nuclear antigen (PCNA), and positive cells were counted by an image analyzer using light microscopy. In cephalic and caudal lobes, a small number of positive cells were observed in controls and molting hens 2 d after cessation of lay. The frequency of PCNA-positive cells started to increase from 7 d after cessation of lay (3 d after refeeding), and a significantly higher frequency of positive cells was observed in both lobes of molting hens 13 d after cessation of lay when compared to control hens, molting hens 2 d after cessation of egg lay, and second cycle laying hens. The frequency of PCNA-positive cells in second cycle laying hens decreased to a similar level to that of control hens. These results suggest that anterior pituitary tissue may be remodeled partially by a proliferation of cells during induced molting in hens. PMID- 9355151 TI - Ornithine decarboxylase activity in muscle, liver, and intestinal tissue of turkeys during a short-term feed withdrawal and following refeeding. AB - The activity of ornithine decarboxylase (ODC), an enzyme associated with cellular growth and protein synthesis, was examined in breast muscle, liver, and intestinal tissues of turkeys during a short-term period of feed withdrawal (FW) and following refeeding. Turkeys from a randombred control line were reared under standard management practices to 3 wk of age in battery brooders. Feed was then withdrawn from FW birds for a 48-h period, after which feed was consumed ad libitum. Control birds consumed feed ad libitum throughout the test period. Tissues were collected from 12 birds per treatment following 24 and 48 h of FW and at 6, 12, 24, and 48 h following refeeding for later determination of tissue ODC activity. Activity of ODC was greater in tissue from the small intestine than in liver tissue and both had greater activity than that exhibited by breast muscle. Short-term FW and refeeding produced differential responses in ODC activity of the three tissues examined. Feed withdrawal resulted in a reduction of ODC activity in intestinal tissue, whereas activity was unaffected for liver or breast muscle tissues. Compensatory increases in ODC activity were observed in liver and intestinal tissues; however, the increase was both more rapid and transitory in small intestine than in liver tissue. The ODC activity in breast muscle was largely unaffected by short-term FW and refeeding. Patterns of ODC activity in liver during FW and refeeding closely resembled patterns observed for absolute and relative liver weight. Thus, the results of the present experiment demonstrate that short-term FW and refeeding influence underlying growth mechanisms of supply organs, such as hepatic and intestinal tissue, in addition to affecting overall growth and muscle development. PMID- 9355153 TI - Effect of dietary aminoguanidine on tissue pentosidine and reproductive performance in broiler breeder hens. AB - Factors influencing the age-related decline in production parameters of broiler breeder hens are poorly understood. Elevated blood glucose concentrations measured in broiler breeder hens may contribute to this decline. The nonenzymatic attachment of glucose to proteins generates glycoxidation crosslinks in tissue proteins, which can ultimately impair their function. One such glycoxidation crosslink, pentosidine, has been used as a biomarker for aging studies because of its accumulation on the structural protein collagen. The objectives of these studies were to determine whether pentosidine accumulates with age in hens and whether the crosslinking inhibitor, aminoguanidine (AG), could retard this accumulation. An additional objective was to determine whether AG had any effect on production performance. In the first study, broiler breeder hens (n = 318) were randomly assigned to two groups: control and supplemented (400 ppm AG). Pentosidine was measured in the skin of the birds at 20 and 67 wk of age. Egg production was measured daily. In a second study, broiler breeder hens (n = 60) were reared as previously described. Pentosidine was measured in the skin of the birds at 20 and 68 wk of age. Results showed that pentosidine was present in the skin of the hens, and that concentrations increased with age (P < 0.001). Although pentosidine was reduced (P < 0.001) in AG-supplemented birds, production performance was not affected. In conclusion, AG retarded the rate of accumulation of pentosidine during lay in broiler breeder hens, but the reduction in pentosidine did not significantly affect production performance. PMID- 9355154 TI - Cell proliferation in the process of oviducal tissue remodeling during induced molting in hens. AB - Tissue remodeling and calcium binding protein-D28K (CaBP-D28K) dynamics were examined in the oviduct relative to induced molting. The oviducal tissues of premolting, 7, 10, 13, and 16 d after cessation of laying, and postmolting hens were examined. Frequency of proliferating cells and immunoreactive CaBP-D28K molecules were identified by immunocytochemistry for proliferating cell nuclear antigen (PCNA) and Western blot for CaBP-D28K. The relative frequency of PCNA positive cells in the mucosal epithelium of the magnum, isthmus, and shell gland was low in premolting and 7 d after the cessation of egg laying. In the magnum and isthmus it was markedly increased in 10 and 13 d after cessation, followed by a slight decrease at 16 d after cessation. The frequency in the shell gland was kept high 10, 13, and 16 d after cessation. The frequency of PCNA-positive cells in each segment was decreased when the birds resumed laying. In the stroma of magnum and isthmus where the tubular glands were located, the frequency of PCNA positive cells was significantly increased 10 d after the cessation of egg laying relative to premolting and 7 d after cessation. In postmolting hens and in hens 16 d after cessation, the frequency was decreased to a same level to that of premolting hen. In the shell gland, the frequency of PCNA-positive cells was high at 10, 13, and 16 d, and diminished in postmolting hen. Single immunoprecipitate band for CaBP-D28K was observed in the shell gland of premolting, postmolting, and younger hens, whereas the density of bands was greater in postmolting hens and younger hens than in premolting hens. We suggest that the oviducal tissues are remodeled by replacing the old glandular cells with new ones that are derived from the mucosal epithelium and uninvoluted glandular cells. Such rejuvenation of shell gland tissue may lead the improvement of CaBP-D28K induction. PMID- 9355156 TI - Feather retention force in broilers ante-, peri-, and post-mortem as influenced by carcass orientation, angle of extraction, and slaughter method. AB - Stunning and slaughter trials were conducted to evaluate the influence of carcass orientation (inverted or supine), angle of feather extraction (parallel or perpendicular to the carcass surface), and slaughter method (exsanguination without or with spinal cord transection) on feather retention force (FRF) in commercial broilers sampled ante-, peri-, and post-mortem. The pectoral, sternal, and femoral feather tracts were sampled before and after stunning contralaterally, with a maximum indicating force gauge, from broilers suspended on a shackle (inverted) or laying on a table (supine). For all trials and sample periods FRF was consistently greater in the femoral area (547 to 679 g) than in the pectoral area (273 to 391 g), with the sternal feather tract requiring the least force at 246 to 343 g. Feathers extracted parallel to the carcass resulted in consistently greater FRF (9 to 29%) than feathers extracted at a perpendicular angle, at all sample periods. Broilers suspended on shackles ante- and peri mortem had higher FRF values (5 to 30%) than those restrained in shackles in a supine position on a table. Other parameters resulted in minor and inconsistent alterations in FRF. Electrical stunning, when not followed by bleeding, resulted in small reductions in FRF (up to 7%). Bleeding after stunning without or with spinal cord transection resulted in variable peri-mortem FRF changes (+7 to -11% and +11 to -11%, respectively). Only in the pectoral feather tract was there a significant increase (5 to 6%) in FRF as broilers went from the ante- to peri mortem state. At 2 and 6 min after stunning and initiation of exsanguination, post-mortem FRF was unaffected by carcass orientation for the pectoral and femoral tracts. PMID- 9355155 TI - Effect of postchill aging and sodium tripolyphosphate on moisture binding properties, color, and Warner-Bratzler shear values of chicken breast meat. AB - The objective of this study was to assess the effects of treating chicken breast forequarters with sodium tripolyphosphate (STPP) after various postchill storage times on meat quality. Sixty-four commercially reared broilers (two replicates of 32 birds each) were slaughtered and chilled, and then the forequarters (split breasts with spine and ribs) were harvested and aged for 0, 120, 180, or 240 min postchill. After each aging period, one forequarter from each of 16 birds was marinated with a NaCl solution and the opposite forequarter was marinated with the same NaCl solution containing STPP. The quarters were then cooked and the following traits measured: marinade absorption, cooking loss, objective color values, and Warner-Bratzler shear values. As aging time prior to marination increased, cooking loss and redness of the cooked meat decreased, but marinade absorption and the color values were unaffected. The STPP treatment increased marinade absorption, decreased cooking losses, and decreased cooked meat redness (P < 0.05). Shear values decreased with aging time for both the control and STPP treated breast meat. When the STPP treatment was applied immediately after carcass chilling, the STPP-treated meat exhibited shear values more than 60% greater than those of the controls (9.14 and 5.69 kg, respectively). Results indicate that time postchill at which further processed products are treated with STPP can have a significant effect on quality, especially cooked product texture. PMID- 9355157 TI - Feather retention force in broilers ante-, peri-, and post-mortem as influenced by electrical and carbon dioxide stunning. AB - Stunning and slaughter trials were conducted to evaluate the influence of stunning method (electrical 50 V alternating current, CO2 gas: 0 to 40% for 90 s or 40 to 60% for 30 s) on feather retention force (FRF) in commercial broilers. Feathers from the pectoral, sternal, and femoral feather tracts were sampled with a force gauge before stunning (ante-mortem) and contralaterally either after stunning (peri-mortem from 0.5 to 4 min) or after stunning and bleeding (post mortem from 2 to 6 min). Prior to stunning, ante-mortem FRF values varied among assigned stunning methods only for the pectoral (7%) feather tract. After stunning, peri-mortem FRF values were higher only for the sternal tract (11% for 40 to 60% CO2 for 30 s); whereas after stunning and bleeding, post-mortem FRF values were lower than ante- or peri-mortem only for the sternal tract (10% lower for 40 to 60% CO2 for 30 s). Peri- and post-mortem FRF values did not differ among stunning methods for the pectoral and femoral feather tracts. Small changes in FRF values occurred from ante-mortem to peri-mortem (-1 to +12%), and from ante-mortem to post-mortem (-2 to +8%) across stunning methods. A significant increase was determined for only the pectoral tract (7%) from ante- to peri mortem across stunning methods. Electrically stunned broilers that were not bled gained weight in excess of the 36 feathers removed (0.16%), apparently due to body surface water pickup during the brine-stunning process, whereas CO2-stunned broilers lost weight due to excretion of cloacal contents (-0.31 to -0.98%). The change in body weight among stunning methods was significant (P < 0.0233). Peri- and post-mortem FRF, in addition to bleed-out body weight loss, were not substantially influenced by electrical or CO2 stunning methods, and, therefore, carcass defeathering efficiency may not differ after scalding. PMID- 9355158 TI - Effect of electron beam irradiation on physical, physiochemical, and functional properties of liquid egg yolk during frozen storage. AB - Raw yolk of 1-d-old eggs was either subjected to linear electron beam irradiation at approximately 2.5 kGy dosage or not processed. Both irradiated and nonprocessed egg yolk samples were stored at -15 C after irradiation. Testing was conducted on 0, 1, 7, 15, 30, and 60 d of storage. Development of storage modulus (G') was delayed in irradiated samples after 7 d, which suggests that less structure was developed in irradiated egg yolk than in nonprocessed egg yolk during storage. Irradiated samples retained more soluble protein within the first 7 d and showed slightly improved emulsion capacity over that from nonprocessed samples. However, irradiated egg yolk was less bright than nonprocessed samples. No differences were observed in SDS-PAGE patterns of soluble proteins and delipidized low density lipoprotein (LDL). The LDL isolated from irradiated liquid egg yolk showed no difference in N-terminal amino acids compared to that of nonprocessed egg yolk, indicating no detectable cleavage of LDL. However, the denaturation temperature of irradiated samples at Day 0 shifted about 1 C lower than that of the nonprocessed sample. Results indicated that electron beam irradiation did not cause significant physical, chemical or functional changes of egg yolk, or cleavage of egg yolk protein. Therefore, electron beam irradiation could serve as a preservation method for liquid egg yolk. PMID- 9355159 TI - The effect of seasonal heat stress on rigor development and the incidence of pale, exudative turkey meat. AB - Heat stress is one of the prominent ante-mortem stressors that elicits pale, soft, and exudative meat characteristics in stress-susceptible pigs. Industry reports of exudative turkey meat increase in the early summer with the onset of prolonged high temperatures. To study the effect of seasonal heat exposure on turkeys, 122 17-wk-old Nicholas tom turkeys were subjected in January either to growth temperatures of 16/24 C (night/day) (control) or to elevated temperatures of 32/38 C (night/day) (heat-stressed, HS). Turkeys were processed at 21 wk of age in a manner simulating commercial conditions. Pectoralis muscle samples were taken at 15 min (prechill), 2 h (postchill), and 24 h and analyzed for R-value, pH, and color. At 2 h, the remaining intact Pectoralis muscle was harvested, aged on ice for 23 h, and analyzed for drip loss and cook loss. Percentage mortality and carcass weights were not significantly different between treatments. By 15 min post-mortem, the HS birds exhibited a faster pH decline and had higher R values that persisted through 24 h. The HS birds were also paler in color and exhibited increased drip loss and cook loss when compared to controls; however, expressible moisture was not different between treatments. In addition, the HS birds had a higher frequency of abnormal birds than controls when birds were grouped as normal (L* < 53) or abnormal (L* > 53). PMID- 9355160 TI - Pooled standard error?? PMID- 9355161 TI - The denial of severe mental illness. PMID- 9355162 TI - Assessment of symptoms of attention-deficit hyperactivity disorder in adults with substance use disorders. PMID- 9355163 TI - Sources of patient-care income, work settings, and age of male and female psychiatrists. PMID- 9355164 TI - Dispelling the stigma of schizophrenia. PMID- 9355165 TI - Examining the link between high-volume providers and shorter inpatient stays. PMID- 9355166 TI - Clinically based risk management principles for recovered memory cases. AB - Controversy over cases involving so-called recovered memories of sexual abuse has threatened to divide the mental health field, just as lawsuits based on recovered memories have sometimes divided children from parents and others. The authors review issues in this controversy, including the role of misdirected advocacy for recovered memory by some practitioners, the distinction between the actual events and patient's narrative truth as a factor in the therapeutic alliance, and the contrast between therapeutic and legal remedies. They recommend nine clinically based risk management principles to guide clinicians in dealing with cases involving recovered memory. They include the need for documentation and consultation; the value of psychotherapeutic neutrality, maintaining a calm perspective, and understanding the difference between historical and narrative truth; the incompatibility of the roles of treater and forensic expert; the risks of special therapies such as hypnosis; awareness of the roles of other professionals and the significance of the patient's family; and the importance of knowing when to end treatment. PMID- 9355167 TI - Utilization of mental health services by women in a male-dominated environment: the VA experience. AB - OBJECTIVE: The study examined whether women in the Veterans Affairs system use mental health services at a lower rate than men because the system is geared to treat a mostly male population. METHODS: Data were obtained on a national cohort of patients utilizing specialty mental health services in the VA during a two week period in fiscal year 1991 (N = 70,979). Analyses included comparison of the proportion of women among treated veterans with the age-adjusted proportion of women among all veterans, comparison across gender of the likelihood of use of any general psychiatric services or substance abuse care in 1991, comparison of the intensity of services used (inpatient days and outpatient contacts) by service users in 1991, and comparison of the likelihood of receiving care and the intensity of mental health services received two years later. RESULTS: Overall, 3.95 percent of veterans who used VA mental health specialty services were women; 4.02 percent of all veterans were women. No significant differences between genders were found in use of general psychiatric services, either in the likelihood of any use or the intensity of services used. However, women were significantly less likely to receive substance abuse care (16.3 percent of women versus 71.2 percent of men); once receiving care, they used a similar intensity of substance abuse services. CONCLUSIONS: Being a woman does not appear to have a substantial effect on overall access to VA mental health services or use of general psychiatric services; however, women use VA substance abuse treatment services at a lower rate than men. PMID- 9355168 TI - The MacCAT-T: a clinical tool to assess patients' capacities to make treatment decisions. AB - OBJECTIVE: The feasibility, reliability, and validity of a new instrument, the MacArthur Competence Assessment Tool-Treatment (MacCAT-T), which was developed for use by clinicians, was tested. The instrument assesses patients' competence to make treatment decisions by examining their capacities in four areas- understanding information relevant to their condition and the recommended treatment, reasoning about the potential risks and benefits of their choices, appreciating the nature of their situation and the consequences of their choices, and expressing a choice. METHOD: The MacCAT-T and instruments to measure symptom severity were administered to 40 patients recently hospitalized with schizophrenia or schizoaffective disorder and 40 matched subjects in the community without mental illness. RESULTS: A high degree of ease of use and interrater reliability was found for the MacCAT-T. Overall, the hospitalized patients performed significantly more poorly than the community subjects on understanding and reasoning, although many patients performed as well as community subjects. Poor performance was related to higher levels of some psychiatric symptoms, such as conceptual disorganization, hallucinations, and disorientation. CONCLUSIONS: The MacCAT-T offers a flexible yet structured method with which caregivers can assess, rate, and report patients' abilities relevant for evaluating competence to consent to treatment. PMID- 9355170 TI - Psychiatric status, quality of life, and level of care as predictors of outcomes of acute inpatient treatment. AB - OBJECTIVE: The study examined the role of five types of variables--demographic characteristics, psychiatric status, functional quality of life, satisfaction with quality of life, and level of care--in predicting key outcomes of inpatient treatment. METHODS: Multivariate canonical regression and univariate multiple regression models were constructed using data from 1,053 inpatients at a public hospital in Washington State. The models were used to predict length of stay, change in symptom severity during hospitalization, psychiatrists' ratings of patients' insight into their illness at discharge, patients' global satisfaction with life, and rehospitalization within 18 months. Hierarchical stepwise procedures were used to select variables that were significant predictors of outcomes. RESULTS: All five classes of predictors were related to the outcomes. The roles of demographic characteristics and diagnoses were minimal. Previous hospitalization and severity of symptoms at admission were strong predictors of psychiatric status. Indicators of functional quality of life and satisfaction with quality of life explained significant variance in all models after accounting for the other classes of predictors. Frequency of family visits was the strongest functional quality-of-life predictor, relating to positive outcomes. Pretreatment satisfaction with life was a significant predictor of most outcomes, and increased satisfaction was associated with positive outcomes. CONCLUSIONS: Patients' quality of life before psychiatric inpatient treatment predicted treatment outcomes independently of psychiatric status, demographic characteristics, and level-of-care variables. Prospective studies are needed to predict outcomes using multidimensional constructs. PMID- 9355169 TI - Effect of antidepressant therapy on health care utilization and costs in primary care. AB - OBJECTIVE: Four groups of patients receiving different antidepressant drugs in a primary care setting were compared in terms of duration of antidepressant therapy and health and mental health care utilization and costs. METHODS: A retrospective analysis of the medical and pharmacy claims of an employed population and their families was conducted. A total of 1,242 patients with a diagnosis of depression were included in the analyses. The four antidepressant cohorts were fluoxetine (N = 799), trazodone (N = 89), the tricyclics amitriptyline and imipramine (N = 104), and the secondary amine tricyclics desipramine and nortriptyline (N = 250). The primary outcome measures were total health care charges, total charges for mental health services, and the pattern of antidepressant use. Secondary measures included charges for outpatient care and pharmacy and the number of outpatient visits. Data analysis involved use of two-stage multivariate regression modeling known as sample selection models. RESULTS: Patients taking fluoxetine achieved higher rates of continuous use for at least six months compared with those taking the other drugs. After selection bias due to observed and unobserved characteristics and other confounding variables was adjusted for, no significant differences were found between drug cohorts in total medical charges. CONCLUSIONS: Improvements in the process of care at no apparent increase in total charges appear possible through appropriate medication therapy. PMID- 9355171 TI - Effectiveness of a continuum of care using brief and partial hospitalization for agitated dementia patients. AB - OBJECTIVE: A behavioral intensive care unit was originally designed as a 21-day inpatient program for treating agitation among demented patients, one of the most common behavioral disorders in this group. Due to the need to dramatically reduce length of stay and create alternative care environments, the original model was modified into an integrated continuum of care blending inpatient and outpatient care and partial hospitalization that reduced hospitalization from 21 to an average of seven days. This quasiexperimental study compared the effectiveness of the inpatient and continuum-of-care programs and conducted cost analyses. METHODS: Subjects were inpatients diagnosed with both dementia and agitation. Outcomes of 68 patients treated in the inpatient program were compared with those of 110 patients treated in the continuum of care. The primary outcome measure was patients' score on the Cohen-Mansfield Agitation Inventory, which provides a total agitation score and scores on three factors describing agitated behavior- physically aggressive behavior, verbally aggressive behavior, and nonaggressive behavior. RESULTS: A statistically significant reduction in agitation was found for patients treated in both programs, with no significant difference in outcome between programs. Patients in both programs showed significant improvements in physical aggression, verbal aggression, and nonaggressive behavior. The cost effectiveness analysis revealed clear advantages for the continuum-of-care program, especially in the area of aggressive behaviors. CONCLUSIONS: The data suggest that the restructured program is an effective and economically feasible intervention. PMID- 9355172 TI - Methods of cost-effectiveness analysis in the assessment of new drugs for Alzheimer's disease. AB - New treatments for Alzheimer's disease highlight the complex clinical and financial issues at stake with new pharmacotherapies. This paper describes cost effectiveness analysis as a method for assessing these issues. Cost-effectiveness analyses show the relationship between resources used and health benefits achieved for a medical intervention compared with an alternative strategy. In analyses of treatments of Alzheimer's disease, costs include health care resources, such as diagnostic tests, medications and efforts to monitor or treat side effects, acute hospital care, physicians' services, home health care, and nursing home care; non-health-care resources, such as support services provided by paid caregivers; and time spent by family members in unpaid provision of care and by patients in seeking care or undergoing an intervention. Effectiveness of interventions can be assessed by measuring changes in patients' cognitive functioning or by measuring years of life gained and the quality of life during those years. Cost-effectiveness studies often make use of disparate data sources, including data collected as part of randomized controlled clinical trials, and they often use mathematical models to support estimates. Because economic evaluations of new interventions for Alzheimer's disease will likely play an increasingly influential role in clinical and resource allocation in the coming years, physicians and other health system stakeholders should familiarize themselves with the techniques of cost-effectiveness analysis and become critical consumers of the literature describing these analyses. PMID- 9355173 TI - Clinicians' referral and matching of substance abuse patients to self-help groups after treatment. AB - OBJECTIVE: The clinical practice guidelines for substance use disorders from the American Psychiatric Association (APA) recommend referral of some patients to self-help groups. The purpose of this study was to determine current patterns of referral to self-help groups in substance abuse treatment programs in the United States and compare them with referral recommendations in APA guidelines. METHODS: Directors of all 389 substance abuse treatment programs in the Department of Veterans Affairs health care system completed a mailed survey on posttreatment self-help referral practices. RESULTS: Survey responses indicated that a large proportion of substance abuse patients were referred to Alcoholics Anonymous (79.4 percent), with other self-help organizations receiving a smaller but significant number of referrals. Referrals to 12-step self-help organizations were more common in programs that endorsed a 12-step treatment orientation and that employed a higher proportion of staff members in recovery from substance use disorders. Consistent with APA practice guidelines, clinicians were less likely to make a referral to a 12-step self-help group if a patient was an atheist, had a comorbid psychiatric disorder, or had less severe substance abuse problems. In deciding whom to refer to self-help groups, clinicians also considered other variables that are not addressed in current practice guidelines, such as age and previous involvement in 12-step groups. CONCLUSIONS: Clinicians make extensive use of self-help groups for their patients, as recommended in APA practice guidelines. However, some differences between current practice and recommended practice warrant further investigation. PMID- 9355174 TI - Gold award. Linking mentally ill persons with services through crisis intervention, mobile outreach, and community education. Project Respond, Mental Health Services West, Portland, Oregon. PMID- 9355175 TI - Summarizing clinical psychiatric data. AB - The authors propose a method of summarizing clinical data to serve patient care. The proposed one-page summary uses diverse visual presentations of data, including small graphs for ratings and drug dosages, time lines for clinic visits and hospital stays, and genograms for inherited illness, as well as a textual presentation of recent clinical notes. The summary depicts a patient's recent and lifetime clinical experience. It allows the viewer to assess relationships between interventions and outcomes for psychiatric and medical problems. Computerized patient information systems, which are increasingly being used, can present data in virtually any form. The authors hope to encourage mental health professionals to reshape psychiatric records. PMID- 9355177 TI - Consumer satisfaction with telemedicine child psychiatry consultation in rural Kentucky. AB - Forty-three rural Kentucky families who obtained child psychiatry consultation during the initial eight months of the University of Kentucky's telemedicine program completed questionnaires assessing their satisfaction with telemedicine. Respondents were 46 parents and nine children. All respondents reported that they were very satisfied with the consultation; all of the children and 98 percent of the parents reported that they were as satisfied with the telemedicine consultation as with an in-person visit. Few respondents reported nervousness about using the equipment. These results suggest that child psychiatry consultation via telemedicine provides high levels of satisfaction for both children and adults. PMID- 9355176 TI - Clinical characteristics and health resource use of men and women veterans with serious mental illness. AB - In this retrospective analysis of gender-specific differences among veterans with serious mental illness, the clinical characteristics and health service utilization of 57 women and 114 men were compared. Women had fewer comorbid psychiatric illnesses than men, and substance use disorders were the most frequent comorbid psychiatric illness for both genders. Unlike nonveteran samples with serious mental illness, the veterans in this study showed no gender differences in hospital length of stay. Atypical antipsychotics, used for only suboptimally responsive illness in the study group, were prescribed for 50 percent of women with primary psychosis, compared with 15.3 percent of men with primary psychosis. The results suggest that psychosis among women veterans is more severe or refractory than that among men veterans. PMID- 9355178 TI - Preventing aggression. PMID- 9355179 TI - Confidentiality as a barrier to treatment. PMID- 9355180 TI - Improving patients' drug compliance. PMID- 9355181 TI - Number of new AIDS cases in U.S. declines for first time since AIDS/HIV epidemic began. PMID- 9355182 TI - Frosted branch angiitis: clinical syndrome or clinical sign? PMID- 9355183 TI - The role of vitreoretinal surgery in the treatment of posttraumatic macular hole. AB - PURPOSE: To determine if vitreoretinal surgery is successful in closing traumatic macular holes and subsequently improving visual acuity. Blunt trauma may result in a macular hole when it causes traumatic separation of the vitreous from the retina, contusion necrosis, or subfoveal hemorrhage. Like idiopathic macular holes, traumatic macular holes are surrounded by a ring of subretinal fluid and result in severely diminished visual acuity. METHODS: Fourteen eyes with full thickness posttraumatic macular holes were treated. The patients ages ranged from 15 years to 36 years (mean, 22 years). Preoperative best corrected visual acuity ranged from 20/200 to 20/50 (mean, 20/80). A pars plana vitrectomy and posterior hyaloid dissection were performed, followed by complete fluid-gas exchange and instillation of 0.1 mL of platelet concentrate just over the macular hole. A final flushing with 25% sulfur hexafluoride was done. The postoperative follow-up period ranged from 6 months to 28 months (average, 13 months). RESULTS: Successful anatomic macular hole closure was achieved 6 months after surgery in 13 years (92.86%). Visual acuity improved four or more lines on the Snellen chart within 6 weeks after surgery in every eye with a closed hole; a final visual acuity of 20/20 was achieved in two eyes (15.3%). The mean postoperative visual acuity was 20/30. No intraoperative or postoperative complications were noted, and the lens remained clear in all eyes during the follow up period. CONCLUSION: Our results suggest that intraoperative application of platelet concentrate in combination with vitrectomy may be useful in managing posttraumatic full thickness macular holes, thus improving anatomic and visual outcomes. The greater recovery of visual acuity obtained in this study compared with that obtained in other series of idiopathic macular holes could be related to the young age of the patients with traumatic macular holes and the early diagnosis and treatment. PMID- 9355184 TI - Laser photocoagulation for macular soft drusen. Updated results. AB - PURPOSE: To update the results of a study on the disappearance of macular soft drusen after laser treatment in the natural evolution of age-related macular degeneration. METHODS: A total of 46 patients with confluent soft drusen and pigmentary changes were studied prospectively. Group 1 was composed of 30 patients with bilateral drusen; the authors randomly assigned one eye of each patient for treatment and the fellow eye for the control. In 16 patients a choroidal neovascular membrane was present in one eye, and treatment was applied to the fellow eye (group 2). Argon green laser treatment was applied directly to the soft drusen in the temporal macula. RESULTS: All treated drusen disappeared in a mean of 3.5 months after treatment, and untreated drusen disappeared in all but three patients in an average of 8.5 months. After an average period of 3 years, only one control eye and none of the treated eyes in group 1 developed a choroidal neovascular membrane (P = 0.500). In group 2, neovascularization occurred in 18% of the patients. The initial improvement in Snellen acuity after subfoveal drusen disappearance diminished as a consequence of cataract progression. CONCLUSIONS: Although no definitive conclusions should be made because of the small number of patients studied, results seem to show that this treatment does not reduce the risk of choroidal neovascularization in the treated eye of patients with a history of exudative disease in the fellow eye. It may be effective in patients with high-risk bilateral soft drusen, that is, in less advanced stages of the disease. PMID- 9355185 TI - Imaging of retinal autofluorescence in patients with age-related macular degeneration. AB - BACKGROUND: Several studies have demonstrated an accumulation of autofluorescent materials in the retinal pigment epithelium (RPE) with increasing age. Histologic analysis revealed that lipofuscin is the most potential fluorescent substance in the aging RPE. Therefore, it has been speculated that lipofuscin precedes the presence of drusen and the development of age-related macular degeneration (ARMD). METHOD: A scanning laser ophthalmoscope was used to visualize autofluorescence in the fundus of patients with early ARMD. The digital recordings were analyzed offline with a digital image analyzing system. RESULTS: In 85 of 103 patients, focal areas of increased autofluorescence were visible. Areas of hyperpigmentation at the level of the RPE showed increased autofluorescence in all cases, whereas areas of depigmentation demonstrated decreased autofluorescence. CONCLUSION: Patients with ARMD demonstrated focal accumulation of fluorescent material most likely lipofuscin. Thus, the scanning laser technique combined with an image analyzing system may help to identify eyes at risk for developing exudative ARMD. PMID- 9355186 TI - Retinal detachment in Marfan's syndrome. Characteristics and surgical results. AB - BACKGROUND: The presence of a rhegmatogenous retinal detachment in a patient with Marfan's syndrome is manifested by narrow pupils, dislocated lenses, and a spectrum of pathology ranging from simple holes to giant tears with or without proliferative vitreoretinopathy. PATIENTS AND METHODS: Thirteen patients (18 eyes) with Marfan's syndrome underwent surgery for retinal detachment. Characteristic findings were a retinal detachment in three or more quadrants (12 eyes), a single tear smaller than 30 degrees (eight eyes), a tear between 80 degrees and 120 degrees (five eyes), equatorial and postequatorial tears (11 eyes), and advanced proliferative vitreoretinopathy (seven eyes). Nine uncomplicated retinal detachments were managed with scleral buckling, and nine complicated retinal detachments were managed with pars plana vitrectomy, scleral buckling, and retinal tamponade, mostly with silicon oil. RESULTS: The results of surgery varied, depending on the nature of the retinal tear and the presence of proliferative vitreoretinopathy. Complete retinal reattachment was achieved in 89% of uncomplicated retinal detachments and in 56% of complicated retinal detachments. Additional partial anatomic success was achieved in two eyes with complicated retinal detachments where the macula was attached. Visual acuity improved significantly in five eyes with uncomplicated retinal detachments (median final vision, 20/80) and in six eyes with complicated retinal detachments (median final vision, 20/200). CONCLUSION: The results of surgical treatment for retinal detachments in patients with Marfan's syndrome were comparable with those in patients without Marfan's syndrome. In seven cases of retinal detachment in patients with Marfan's syndrome, we were able to reattach the retina without removing the dislocated intraocular lens. PMID- 9355187 TI - Effect of head position on refraction in aphakic and phakic silicone-filled eyes. AB - BACKGROUND: The refraction of the eye is altered significantly after the instillation of silicone oil into the vitreous cavity because of the silicone oil's high refractive index. The degree of the refractive change shows diurnal variation and depends on the patient's head position. PATIENTS AND METHODS: To analyze the degree and time course of refractive changes with respect to head position, the authors performed objective refraction in five aphakic and five phakic silicone oil-instilled eyes with attached maculae using an automatic hand held refractometer. In each patient, 25 measurements were taken at short intervals using a hand-held autorefractometer with the patient in different positions (supine, head tilted down, and primary eye position). RESULTS: The highest shift of refraction was +5.95 +/- 2.63 diopters spherical equivalent in aphakic eyes and +2.45 +/- 0.71 diopters in the phakic eyes after the patient's position was changed from supine to head tilted down. Three minutes after the position change the refractive shift was stable in almost all eyes. Only slight changes in cylinder and the respective corresponding axis with a mean cylinder shift of 10.1 +/- 5.1 degrees were noted. CONCLUSIONS: Immediately after the head position was changed, aphakic eyes demonstrated more pronounced refractive shifts than phakic eyes. The results of this study explain patients' reports of diurnal changes in visual acuity after intraocular silicone oil instillation. Refractive changes caused by silicone oil stabilized a few minutes after a change in head position. PMID- 9355188 TI - Gyrate atrophy-like phenotype with normal plasma ornithine. AB - PURPOSE: To describe the clinical characteristics of a chorioretinal disease with a gyrate atrophy-like phenotype and normal plasma ornithine. METHODS: One family with three men who had progressive chorioretinal disease and three additional patients with simplex cases were examined clinically and with standard electroretinography, electrooculography, and dark adaptometry. RESULTS: In the family, a 70-year-old man and his two sons (39 and 41 years of age) were affected. On ophthalmoscopy, sharply demarcated peripheral patches of retinal pigment epithelium and choroidal atrophy were seen to progress to the posterior pole in the father's eye. In three unrelated men (62, 70, and 80 years of age), chorioretinal atrophy was present in the mid- and far periphery. Visual acuity was normal in the two youngest of all six patients; however, electroretinogram and electrooculogram waves were reduced. Advanced visual field defects and visual acuity loss occurred in the four older patients. Electroretinogram and electrooculogram were reduced, and the dark adaptation thresholds were elevated. In all patients, serum ornithine levels were normal. Ornithine-delta aminotransferase activity in cultured skin fibroblasts and the apparent Michaelis constant (Km) for ornithine and alpha-ketoglutarate were within the normal range in all patients. CONCLUSIONS: A gyrate atrophy-like phenotype can result from causes other than deficient ornithine-delta-aminotransferase. Its occurrence in three male members in two generations in one family suggests an autosomal dominant inheritance in at least some such patients. PMID- 9355189 TI - Transscleral retinopexy using a continuous wave Nd:YAG laser. AB - PURPOSE: Intraoperative retinopexy currently is performed transsclerally during surgery for retinal detachment by means of cryocoagulation, which usually results in much damage to the tissue. In this study, we investigated the potency of transscleral chorioretinal laser photocoagulation in achieving retinopexy with less breakdown of the blood-retina barrier and resulting in decreased protein leakage and reduced dispersion of the retinal pigment epithelium. METHODS: A continuous wave (CW) Nd:YAG laser with a custom-made optical tip was used. The tip consists of an optical fiber protected by a steel casing for indentation of the sclera. Eight patients underwent the procedure. RESULTS: Three to six pulses of 5.2-6.6 W and exposure duration of 300-500 mesec per spot were used to obtain mild to moderate photocoagulation; no retinal complications were observed. An average of 45 coagulations were obtained per patient. One patient showed mild scleral thermal effects. All retinas were attached after surgery, and seven remained attached at the 6-month follow-up examination. CONCLUSION: Transscleral chorioretinal irradiation with the CW Nd:YAG laser is a suitable method for performing retinopexy. Because no thermal damage to the sclera occurred, this method seems to be a safe and effective procedure. PMID- 9355190 TI - Peripheral transscleral retinal diode laser for rubeosis iridis. AB - PURPOSE: To evaluate the results of peripheral transscleral retinal diode photocoagulation with or without transscleral cyclodiode therapy in patients with rubeosis iridis with or without elevated intraocular pressure and no fundal view. METHODS: Peripheral transscleral retinal diode photocoagulation was performed in 15 eyes of 13 patients in an attempt to promote regression of rubeosis. The fundus could not be seen in any of the 15 eyes, so conventional panretinal photocoagulation was not possible. Nine eyes had associated elevated intraocular pressure and were treated with concurrent transscleral diode cyclophotocoagulation. RESULTS: All eyes showed regression of rubeosis. Of the nine eyes treated with combination therapy, six had stabilized intraocular pressure, and three developed hypotony. None of the eyes developed a peripheral retinal detachment, and one eye lost the ability to perceive light. CONCLUSIONS: This method is effective in treating patients with rubeosis iridis when the view of the fundus is inadequate for conventional panretinal photocoagulation and more extensive intraocular surgery is precluded. It may be combined with transscleral cyclophotocoagulation therapy to manage concurrent high intraocular pressure in rubeotic glaucoma, but this involves a risk of postoperative hypotony. PMID- 9355191 TI - Influence of the cannulated vitrectomy system on the occurrence of iatrogenic sclerotomy retinal tears. AB - PURPOSE: To determine whether the use of the cannulated vitrectomy system decreases the incidence of sclerotomy-related retinal tears relative to traditional vitrectomy techniques. METHODS: Forty-one eyes of 77 patients in this study were randomly selected to undergo treatment with the cannulated port system. RESULTS: We demonstrated a statistically significant decreased incidence of sclerotomy tears in the cannulated group relative to the noncannulated group (1% vs. 7.7%, P < 0.05). The benefit of the cannulated port system appears to be greatest in cases in which an inexperienced surgeon is learning the techniques of vitreoretinal surgery, in eyes with a preoperative diagnosis of tractional diabetic detachment, and in surgery requiring membrane delamination (simple and extensive). CONCLUSION: The three-port vitrectomy system decreases the incidence of sclerotomy-related retinal tears. PMID- 9355192 TI - Lack of p53 protein immunoreactivity in bilateral diffuse uveal melanocytic proliferation. AB - BACKGROUND: Bilateral diffuse uveal melanocytic proliferation is a poorly understood disorder characterized by the progressive proliferation of uveal melanocytes associated with a systemic nonocular malignancy. Overexpression of p53 protein plays a role in the loss of regulatory control of normal cell proliferation, and p53 is the most commonly identified oncogenic protein in human malignancies. We tested the hypothesis that the aberrant cellular activity in bilateral diffuse uveal melanocytic proliferation involves the overexpression of p53 protein. METHODS: Eight eyes from four patients with bilateral diffuse uveal melanocytic proliferation were tested for p53 protein using an immunoperoxidase technique with an anti-p53 protein monoclonal antibody sensitive for normal and mutant p53 protein. RESULTS: The p53 protein could not be detected in any of the eight eyes. CONCLUSIONS: The proliferation of uveal melanocytes in bilateral diffuse uveal melanocytic proliferation does not depend on the overexpression of p53 protein. The loss of cellular regulatory control in bilateral diffuse uveal melanocytic proliferation is probably mediated through another mechanism. PMID- 9355193 TI - Chorioretinitis secondary to mycobacterium tuberculosis in acquired immune deficiency syndrome. AB - BACKGROUND: Several opportunistic intraocular infections have been described in patients with the human immunodeficiency virus, among them infections caused by Mycobacterium tuberculosis. In most cases, the diagnosis is based on clinical findings. Recent reports have described the usefulness of polymerase chain reaction techniques in the diagnosis of bacterial infections. METHODS: The authors observed a 29-year-old woman with acquired immune deficiency syndrome in whom unilateral chorioretinitis developed. The chorioretinitis appeared after cessation of treatment for pulmonary tuberculosis. We obtained aqueous humor by paracentesis and tested it by polymerase chain reaction to detect M. tuberculosis DNA. RESULTS: The polymerase chain reaction of the aqueous humor was positive for M. tuberculosis DNA. CONCLUSION: Polymerase chain reaction was useful in identifying M. tuberculosis in aqueous from a patient with chorioretinitis, pulmonary tuberculosis, and acquired immune deficiency syndrome. PMID- 9355194 TI - Diagnostic and therapeutic challenges. PMID- 9355195 TI - Diagnostic and therapeutic challenges. PMID- 9355196 TI - Lupus associated frosted branch periphlebitis and exudative maculopathy. PMID- 9355197 TI - Steroid-responsive retinal vasculitis with a frosted branch appearance in Crohns disease. PMID- 9355198 TI - Bilateral choroidal effusions and angle-closure glaucoma associated with human immunodeficiency virus infection. PMID- 9355199 TI - Bartonella henselae-associated acute multifocal retinitis in a patient with acquired immunodeficiency syndrome. PMID- 9355200 TI - Ciliochoroidal nerve sheath tumor simulating a malignant melanoma. PMID- 9355201 TI - Exogenous Aspergillus niger endophthalmitis in a patient with a filtering bleb. PMID- 9355202 TI - Sustained-release ganciclovir implants represent an effective treatment option for many patients with cytomegalovirus (CMV) retinitis. PMID- 9355203 TI - Therapeutic efficacy of intravitreal ceftazidime, imipenem, and amikacin in a swine model of posttraumatic Pseudomonas aeruginosa endophthalmitis. PMID- 9355204 TI - A new light source for vitreous surgery. PMID- 9355205 TI - Seronegative spondyloarthropathy in familial Mediterranean fever. AB - To define a possible association between familial Mediterranean fever (FMF) and seronegative spondyloarthropathy (SNSA) and to study features of SNSA in FMF patients, we screened for the presence and manifestations of SNSA in 3,000 FMF patients attending the National Center for FMF in our institution. This population included 160 patients with chronic arthritis, most who suffered from SNSA. Patients were considered to suffer from SNSA if they had chronic arthritis, inflammatory back/neck pain, and sacroiliitis. Patients who had other diseases associated with SNSA were excluded. Eleven patients, nine men and two women, with chronic monoarthritis or oligoarthritis, grade 2 (four patients) or grades 3 to 4 (seven patients), sacroiliitis, and inflammatory back pain met the criteria for diagnosis of SNSA of FMF. These patients were rheumatoid factor (RF) and HLA-B27 negative. In seven patients, spondyloarthropathy developed while they received colchicine, and in four before colchicine. Most patients responded to treatment with nonsteroidal antiinflammatory drugs, but three required second-line agents. These findings suggest that SNSA is one of the musculoskeletal manifestations of FMF that may occur despite colchicine therapy and requires specific treatment. PMID- 9355206 TI - Cost-of-illness of scleroderma: the case for rare diseases. AB - OBJECTIVE: To determine the societal costs of scleroderma (SSc), a rare chronic connective tissue disease that affects approximately 98,000 Americans. Lack of reliable national databases limit rare disease cost studies, and this study suggests methods of using multiple data sources to assess the costs of rare diseases. METHODS: Primary and secondary data sources were used to calculate direct and indirect costs of SSc, including discounted lifetime mortality and morbidity costs. A prevalence-based, human capital approach was used. Sensitivity analyses were used to vary parameters that are uncertain, such as prevalence, mortality, and labor costs. RESULTS: Annual direct and indirect costs of SSc in the United States are $1.5 billion. Morbidity represents the major cost burden, with costs of $819 million (56%) of total costs. The current value of lifetime earnings lost was $179 million (12%) or $300,000 per death. Direct costs were $462 million (32%) or $4,731 per person annually, indicating that costs are spread over the long disease duration. CONCLUSIONS: This study provides one model for the assessment of rare disease costs. Triangulation of data sources and sensitivity analyses are important for determining the costs of rare diseases. The high cost of SSc, despite its low prevalence, suggests that the burden of rare chronic diseases can be high. The high morbidity costs reflect the young age of onset of the disease as well as the need for treatments to decrease morbidity costs. Local shared databases and national surveys are needed to improve cost estimates of rare diseases. PMID- 9355207 TI - Dietary n-3 fatty acids and therapy for rheumatoid arthritis. AB - OBJECTIVE: To examine the potential for dietary n-3 fats to be component of therapy for rheumatoid arthritis (RA). METHODS: Studies of encapsulated fish oil use in RA were reviewed and critiqued, and possible biochemical mechanisms for fish oil effects were examined. The potential for use of n-3 fats was evaluated within a dietary framework rather than a quasi-pharmaceutical framework. RESULTS: There is consistent evidence from double-blind, placebo-controlled clinical trials that dietary n-3 fats, supplied as fish oil, can have beneficial effects in RA. The beneficial effects appear modest, but their size and extent may have been moderated by common trial design factors such as high n-6 polyunsaturated fat diets and concurrent antiinflammatory drug use. Mechanisms for the clinical effects of n-3 fats in RA may involve their ability to suppress production of inflammatory mediators, including n-6 eicosanoids and proinflammatory cytokines. Suppression of n-6 eicosanoid and cytokine production will be possible using foodstuffs that are rich in n-3 fats and poor in n-6 fats. CONCLUSIONS: There are many overlapping biochemical effects of n-3 fatty acids and antiinflammatory pharmaceuticals that could explain the clinical actions of n-3 fats in RA. They suggest that there is the potential for complementarity between drug therapy and dietary choices that increase intake of n-3 fats and decrease intake of n-6 fats. In particular, there is the potential for drug-sparing effects. Future studies with n-3 fats in RA need to address the fat composition of the background diet and the issue of concurrent drug use. PMID- 9355209 TI - Efficacy of multidisciplinary team care programs in rheumatoid arthritis. AB - OBJECTIVE: To assess the efficacy of multidisciplinary team care programs in rheumatoid arthritis (RA). METHODS: Data were obtained by a Medline and a manual search of the literature through January 1997. Both the design and analysis aspects of controlled trials were evaluated. RESULTS: Forty-two papers reporting on 35 clinical trials of multidisciplinary team care were initially identified. Fifteen trials had a controlled design, nine of which were randomized. Patient characteristics, interventions, end point measures, and presentation of the data varied widely among the controlled studies. In 12 trials, inpatient (n = 6) or outpatient (n = 6) multidisciplinary programs were compared with regular outpatient care. Inpatient programs (average duration, 10 to 28 days) had a direct favorable effect on disease activity, lasting up to 1 year. The effect of outpatient programs (average duration, 1 to 2 years) was less marked, with greater improvement of functional status at the end of the treatment program shown in one study. In three trials, inpatient multidisciplinary programs were compared with similar outpatient programs. One study showed that inpatient care was more effective, whereas in two studies similar results were obtained in both groups. CONCLUSION: Favorable effects on disease activity were seen in most trials comparing short inpatient team care with regular outpatient care. Proof of efficacy of prolonged outpatient team care is scanty. Results of trials comparing inpatient with outpatient team care remain inconclusive. PMID- 9355208 TI - Specificity, pathogenecity, and clinical value of antiendothelial cell antibodies. AB - OBJECTIVE: To characterize the putative target antigens for antiendothelial cell antibodies (AECA), the possible pathophysiological role of AECA, and the clinical value of these antibodies as markers of disease activity. METHODS: A structured literature search was done using Medline in combination with a manual search. Two physicians reviewed all articles of special interest. RESULTS: AECA are a heterogenous group of antibodies directed against a variety of antigen determinants on endothelial cells (EC). The EC antigens can be constitutively expressed, constitutively expressed and modulated by cytokines, or cryptic. In addition, antigen determinants for AECA may also be molecules that adhere to EC ("planted" antigens). However, many AECA antigens are currently not well characterized. AECA are detected in a wide variety of inflammatory disorders. Although probably of limited value in disease diagnosis, the detection of these antibodies may be valuable in following disease activity. In several diseases such as systemic lupus erythematosus and systemic vasculitis, high AECA titers are found during active disease whereas lower titers or disappearence of AECA have been reported during remission. The correlation between changes in AECA titers and disease activity suggests an important role for AECA in processes in which vessel wall damage occurs, although it does not exclude the possibility that AECA are an epiphenomenon of vascular injury. Several recent in vitro studies support a role of AECA in the pathophysiology of these inflammatory disorders. AECA may play a role in the pathophysiology by inducing activation of EC resulting in upregulation in the expression of endothelial adhesion molecules and/or secretion of chemoattractants and cytokines. An alternative mechanism by which AECA could be a trigger in the pathogenesis of some diseases is complement dependent cytotoxicity (CDC) and/or antibody dependent cellular cytotoxicity (ADCC). In experimental animal models, antibodies to antigenic determinants expressed on EC were capable of inducing vascular injury. CONCLUSION: AECA represent a heterogenous group of antibodies directed against a variety of antigenic determinants on EC. They are present in a variety of inflammatory disorders. The detection of these antibodies may be valuable in following disease activity. Further characterization of putative antigens is needed to better understand their pathophysiological role. PMID- 9355210 TI - Epidemiological and clinical aspects relating to the variability of rheumatoid arthritis. AB - OBJECTIVE: To review epidemiological studies dealing with the temporal and geographic variability in the occurrence of rheumatoid arthritis (RA) and clinical studies that address the variability of severity and manifestations among populations. METHODS: An extensive search of the literature, including a Medline search, was completed. Studies addressing the origin, history, and trends in the occurrence of RA were reviewed first. Next, studies of the prevalence and incidence of RA in different populations were reviewed, and occurrence rates compared. Standardization was attempted by tabulating adult prevalence rates of studies using equivalent sets of criteria. Studies comparing RA patients from two populations were sought next. Finally, studies dealing with explanations of the presumed variability were reviewed. RESULTS: Temporal variability is indicated by paleopathological evidence that RA has existed in the New World since 4000 BC, whereas there is no evidence that it occurred in Europe before the 17th century, or in Africa before the 20th century. Epidemiological studies show a possible trend of decreasing incidence of RA in the United States and Western Europe, whereas reports from Africa note a rising incidence. In white populations of Europe and America, prevalence is approximately 1%, and incidence is 0.03%. Significantly higher rates are found in some North American Indians, and significantly lower rates in some Asian and African populations, even when the different population structures are taken into account. In the latter populations, different patterns of occurrence from those observed in whites emerge, such as greater female preponderance and a much younger peak age at onset. Direct standardized comparisons of two diverse populations of RA patients showed some differences in expression, severity, or manifestations of RA between populations. CONCLUSION: The occurrence and manifestations of RA are temporally and geographically variable. PMID- 9355211 TI - Method guidelines for systematic reviews in the Cochrane Collaboration Back Review Group for Spinal Disorders. PMID- 9355212 TI - Classics from the spine literature revisited: a randomized trial of 2 versus 7 days of recommended bed rest for acute low back pain. AB - STUDY DESIGN: Review of a trial of bed rest for patients with acute low back pain. OBJECTIVES: To assess the validity and results of the study, and their applicability to and influence on current clinical practice and recommendations. SUMMARY OF BACKGROUND DATA: Although bed rest has been a cornerstone of treatment for acute low back pain, historically this recommendation was largely based on "expert opinion." In 1986, Deyo et al. published a randomized study of 2 versus 7 days of recommended bed rest for acute low back pain. Despite results from this and other studies, current clinical practice and treatment recommendations continue to overemphasize bed rest. METHODS: The study was reviewed using structured criteria adopted from the medical literature that focus on the validity of the study design, the results of the treatment, and the relevance of the findings to clinical practice. RESULTS: Two hundred and three patients were randomized to 2 versus 7 days of recommended bed rest. Groups were similar at baseline evaluation. Outcomes assessed at 3 and 12 weeks were similar between groups, except that patients receiving a recommendation for 2 days of bed rest had significantly fewer days of work absence than those recommended 7 days. Limitations of the study included poor compliance with recommended bed rest, especially in the 7-day group, a marginal sample size without information on relevant confidence intervals, and patient characteristics that may have affected the generalizability of these findings to others with acute low back pain. CONCLUSIONS: Despite limitations, this study provided strong evidence that less bed rest was associated with similar outcomes for acute low back pain along with quicker return to work. Results from this and other studies support a shift away from bed rest as a primary recommendation in the initial management of low back pain. In spite of this, bed rest recommendations for episodes of low back pain remain common. Additional efforts are needed to change clinical practice. PMID- 9355213 TI - Anatomic relations between the lumbar pedicle and the adjacent neural structures. AB - STUDY DESIGN: The authors analyzed anatomic parameters between the lumbar pedicles and the dural sac as well as the spinal nerve roots. OBJECTIVES: To determine quantitatively the anatomic relations between the lumbar pedicle and adjacent dural sac and nerve roots. SUMMARY OF BACKGROUND DATA: Posterior transpedicular screw fixation is the most commonly used method of instrumentation for stabilization of the unstable lumbar spine. A thorough knowledge of the unique anatomy of the lumbar pedicle and adjacent neural structures may avoid or minimize neurologic complications with pedicular screw placement. METHODS: Fifteen adult cadavers were obtained to evaluate quantitatively the anatomic relations of the lumbar pedicle to the adjacent neural structures. After removal of the laminas and facets, the lumbar pedicles, dural sac, and nerve roots were exposed. Direct measurements were taken from the pedicle to the dural sac medially, to the nerve roots superiorly and inferiorly, and between the pedicles. Also, the superoinferior diameter of the nerve root and the frontal angle of the nerve root were measured. Symmetric structures were measured bilaterally. RESULTS: No consistent changes from L1 to L5 were found in all parameters. The mean distances from the lumbar pedicle to the dural sac medially and to the adjacent nerve roots superiorly and inferiorly for all levels were 1.5 mm, 5.3 mm, and 1.5 mm, respectively. The mean interpedicular distance ranged from 23.2 to 24.4 mm. The mean superoinferior diameter of the nerve root ranged from 3.8 to 4.6 mm. The mean nerve root angle ranged from 33.7 degrees to 39.2 degrees. CONCLUSIONS: On the basis of this study, improper placement of the pedicle screw medially or caudally in the lumbar spine should be avoided. PMID- 9355214 TI - Innervation of "painful" lumbar discs. AB - STUDY DESIGN: The authors investigated the innervation of discographically confirmed degenerated and "painful" human intervertebral discs. OBJECTIVE: To determine the type and distribution patterns of nerve fibers present in degenerated human intervertebral discs. SUMMARY OF BACKGROUND DATA: The innervation of intervertebral discs has previously been extensively described in fetal and adult animals as well as humans. However, little is yet known about the innervation of severely degenerated human lumbar discs. The question may be posed whether a disc that has been removed for low back pain possesses an increased innervation compared with normal discs. METHODS: The presence of nerve fibers was investigated using acetylcholinesterase enzyme histochemistry, as well as neurofilament and substance P immunocytochemistry. From 10 degenerated and 2 control discs, the anterior segments were excised and their nerve distribution studied by examining sequential sections. RESULTS: In all specimens, nerve fibers of different diameters were found in the anterior longitudinal ligament and in the outer region of the disc. In 8 of 10 degenerated discs, fibers were also found in the inner parts of the disc. Substance P-immunoreactive nerve fibers were sporadically observed in the anterior longitudinal ligament and the outer zone of the anulus fibrosus. CONCLUSIONS: Findings indicate a more extensive disc innervation in the severely degenerated human lumbar disc compared with normal discs. The nociceptive properties of at least some of these nerves are highly suggested by their substance P immunoreactivity, which provides further evidence for the existence of a morphologic substrate of discogenic pain. PMID- 9355215 TI - Use of electromagnetic fields in a spinal fusion. A rabbit model. AB - STUDY DESIGN: The biomechanical and histologic characteristics of posterolateral spinal fusion in a rabbit model with and without the application of a pulsed electromagnetic field were analyzed in a prospective, randomized trial. In addition, fusion rate with and without a pulsed electromagnetic field in this model was assessed by biomechanical testing, radiographs, and manual palpation. OBJECTIVES: To evaluate the influence of a pulsed electromagnetic field on the spinal fusion rate and biomechanical characteristics in a rabbit model. SUMMARY OF BACKGROUND DATA: Previous studies performed to assess the benefits of a pulsed electromagnetic field in spinal fusion have been complicated by the use of instrumentation, and the animal models used do not have a pseudarthrosis rate comparable to that seen in humans. In contrast, the posterolateral intertransverse process fusion in the rabbit is uncomplicated by the use of instrumentation and has been shown to have a pseudarthrosis rate similar to that found in humans (5-35%). METHODS: Ten New Zealand white rabbits each were randomly assigned to undergo spinal fusion using either 1) autologous bone with electromagnetic fields, or 2) autologous bone without electromagnetic fields. A specially designed plastic constraint was used to focus the pulsed electromagnetic field over the rabbits' lumbar spine 4 hours per day. Animals were killed at 6 weeks for biomechanical and histologic testing. RESULTS: The rate of pseudarthrosis, as evaluated radiographically and manually in a blinded fashion, decreased from 40% to 20% with the pulsed electromagnetic field, but this decrease in the nonunion rate was not statistically significant given the number of animals per group. Biomechanical analysis of the fusion mass showed that a pulsed electromagnetic field resulted in statistically significant increases in stiffness (35%), area under the load-displacement curve (37%), and load to failure of the fusion mass (42%). Qualitative histologic assessment showed increased bone formation in those fusions exposed to a pulsed electromagnetic field. CONCLUSIONS: This study demonstrates the reproducibility of a rabbit fusion model, and the ability of a pulsed electromagnetic field to induce a statistically significant increase in stiffness, area under the load displacement curve, and load to failure of the fusion mass. This investigation provides a basis for continued evaluation of biologic enhancement of spinal arthrodesis with the use of a pulsed electromagnetic field. PMID- 9355216 TI - Fiber transformations in multifidus muscle of young patients with idiopathic scoliosis. AB - STUDY DESIGN: In this study, the authors investigated the superficial multifidus muscle in patients with idiopathic scoliosis. During spinal fusion, biopsies were taken bilaterally at the apex of the curve, and at the upper and lower end vertebrae. OBJECTIVES: To analyze the muscular reactions in response to bracing in patients with idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: The extent to which intervertebral mobility is restricted by an orthosis is still controversial. In addition, the effect of bracing on the erector spinae has not been investigated. METHODS: Of a total 30 patients, 11 had been treated with a corset for a year or more before surgery. Biopsies were investigated histochemically and the muscle fibers classified as Type I, IIA, IIB, or IIC (transitional fibers). The relative distribution of the fibers was calculated and their diameter was measured. RESULTS: In unbraced patients, a shift in the fiber distribution (from "slow" to "fast") was observed exclusively at the concave side of the apex. This shift was paralleled by an increased percentage of the intermediate Type IIC fiber (indicative of fiber transformation). In patients who always wore a corset, the relative amount of Type IIC fibers was increased, without preference for a specific location. CONCLUSIONS: Corset treatment elicits muscle fiber transformation processes at different levels along the scoliosis. This general reaction of the paraspinal muscles provides strong evidence against the existence of muscular disorders that are restricted to the area of the apex and are thus causing the scoliosis. As such, it must be assumed that the muscular changes in the apical region are secondary. PMID- 9355217 TI - Are sheep spines a valid biomechanical model for human spines? AB - STUDY DESIGN: Range of motion, neutral zone, and stiffness parameters of the complete cervical, thoracic, and lumbar sheep spine were determined in flexion and extension, axial left/right rotation, and right/left lateral bending. OBJECTIVES: To determine quantitative biomechanical properties of the sheep spine and compare them with those from the human spine. SUMMARY OF BACKGROUND DATA: Sheep spines often serve as a model for experimental in vivo and in vitro studies in spine research, but few quantitative biomechanical data from sheep spines for comparison with human specimens are available. METHODS: Complete spines were sectioned into single-joint segments and tested in a spine tester under pure moments in the three main anatomic planes. RESULTS: The craniocaudal variation in range of motion in all load directions was qualitatively similar between sheep spines and values reported in the literature for human specimens. CONCLUSIONS: Based on the biomechanical similarities of sheep and human spines demonstrated in this study, it appears that the use of the sheep spine, which already includes evaluation of surgical techniques and bone healing processes, might be extended to spinal implants. PMID- 9355218 TI - Biomechanical comparison of posterior lumbar interbody fusion cages. AB - STUDY DESIGN: Cadaveric human and bovine lumbar spine models simulating the acute postoperative period were used to compare the biomechanical properties of two designs of intervertebral body threaded fusion cages. The instrumented spines were compared with intact spines and with spines with resected posterior elements, representing a revision case. OBJECTIVE: To determine the relative biomechanical performance of these competing devices. SUMMARY OF BACKGROUND DATA: These cages are currently under clinical investigation, and basic biomechanical data are needed. METHODS: Insertion torques and maximum pushout loads were measured for each cage. Intact spines, posteriorly instrumented spines (posterior lumbar interbody fusion), and spines with resected posterior elements were loaded in axial compression, flexion and extension bending, and axial torsion. Stiffness comparisons were made between the different configurations. RESULTS: Insertion torques and pushout loads were similar for the cages. Both cages significantly increased stiffnesses above those of the intact spines and the resected spines. The BAK-instrumented spines were more stiff in axial compression, while the Threaded Interbody Fusion Device spines were more stiff in extension. CONCLUSIONS: This study revealed the two cages to have similar biomechanical characteristics immediately after posterior insertion and warrant further clinical studies. PMID- 9355219 TI - The effects of padded surfaces on the risk for cervical spine injury. AB - STUDY DESIGN: This is an in vitro study comparing cervical spine injuries produced in rigid head impacts and in padded head impacts. OBJECTIVES: To test the hypothesis that deformable impact surfaces pose a greater risk for cervical spine injury than rigid surfaces using a cadaver-based model that includes the effects of the head and torso masses. SUMMARY OF BACKGROUND DATA: It is widely assumed that energy-absorbing devices that protect the head from injury also reduce the risk for neck injury. However, this has not been demonstrated in any experimental or epidemiologic study. On the contrary, some studies have shown that padded surfaces have no effect on neck injury risk, and others have suggested that they can increase risk. METHODS: Experiments were performed on 18 cadaveric cervical spines to test 6 combinations of impact angle and impact surface padding. The impact surface was oriented at -15 degrees (posterior impact), 0 degree (vertex impact), or +15 degrees (anterior impact). The impact surface was either a 3-mm sheet of lubricated Teflon or 5 cm of polyurethane foam. RESULTS: Impacts onto padded surfaces produced significantly larger neck impulses (P = 0.00023) and a significantly greater frequency of cervical spine injuries than rigid impacts (P = 0.0375). The impact angle was also correlated with injury risk (P < 0.00001). CONCLUSIONS: These experiments suggest that highly deformable, padded contact surfaces should be used carefully in environments where there is the risk for cervical spine injury. The results also suggest that the orientation of the head, neck, and torso relative to the impact surface is of equal if not greater importance in neck injury risk. PMID- 9355220 TI - Bone mineral density in patients with ossification of the posterior longitudinal ligament. Minimal decrease of bone mineral density with aging. AB - STUDY DESIGN: Bone mineral density of individuals with ossification of the posterior longitudinal ligament and that of normal people was determined by dual energy x-ray absorptiometry. OBJECTIVES: To determine whether bone mineral density in the people with ossification of the posterior longitudinal ligament is higher than that in normal individuals even in body parts other than the spine, and to evaluate the relation between bone mineral density and age in patients with ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: It is unknown whether the bone mineral density of patients with ossification of the posterior longitudinal ligament is greater in body parts other than the spine. If so, it provides a basis for the theory that certain systemic factors are involved in the pathogenesis of ossification of the posterior longitudinal ligament. Because bone mineral density decreases physiologically after middle age, the influence of age must be considered in evaluating bone mineral density. METHODS: In the rib area and upper and lower limb areas, which are not affected by ossification of the spinal ligament, bone mineral density of 45 men with ossification of the posterior longitudinal ligament of the cervical spine was compared with that of 25 men without ossification of the posterior longitudinal ligament (normal group). RESULTS: Bone mineral density was higher in the group with ossification of the posterior longitudinal ligament in each part and significantly higher in the rib and lower limb areas (rib: P < 0.01, lower limb: P < 0.05). The age-related decrease was significantly less in the group with ossification of the posterior longitudinal ligament (rib: P < 0.01, upper limb: P < 0.05, lower limb: P < 0.01). CONCLUSIONS: Systemic factors that increase bone mineral density appear to be involved in the pathogenesis of ossification of the posterior longitudinal ligament, and these factors may be activated after middle age. PMID- 9355221 TI - Regional variation in vertebral bone density and trabecular architecture are influenced by osteoarthritic change and osteoporosis. AB - STUDY DESIGN: The effects of age-related osteoarthritic disease and bone loss on cortical and trabecular bone density in the human spine were analyzed. Changes were quantified by a new method of computed quantitative radiography. Using this method, the relative losses of bone mineral from specific areas and the specific redistribution of bone within vertebrae as a function of age-related bone loss and osteoarthritic change were determined. OBJECTIVES: To quantify the effects of age-related bone loss and osteoarthritic disease on cortical and trabecular density in the human spine by the use of a new method of computed quantitative radiography. SUMMARY OF BACKGROUND DATA: To the authors' knowledge, osteoarthritic and age-related changes resulting in the relative loss of bone mineral from specific areas within vertebrae have never been quantitatively studied. METHODS: Seventy-four vertebrae were obtained from 23 normal, human subjects at autopsy. Vertebral bodies were measured by dual-energy x-ray absorption, and then sectioned for analysis by computerized quantitative radiography. Attention was focused on overall bone density, regional cancellous bone density, and cortical bone density (anterior cortex, superior, and inferior endplates) for both mid-sagittal and para-sagittal sections. One hundred forty sections were so analyzed. RESULTS: Quantitative radiography demonstrated previously undescribed, well defined patterns of cortical and trabecular architecture and bone density within vertebral bodies that were uniform among vertebrae, but strongly influenced by osteoarthritic change and bone loss. These changes were neither detected nor documented by dual-energy x-ray absorption. CONCLUSIONS: Understanding the patterns of bone lose, and knowing how osteoarthritic change affects normal bone loss yields insight into the pathophysiology of the aging process and osteoarthritic disorders. Knowledge of the variations in density and microanatomy within vertebrae may improve techniques of internal fixation and designs of spinal implants, and may help to clarify the pathogenesis of clinical problems such as fractures. PMID- 9355222 TI - Measurement of vertebral rotation in adolescent idiopathic scoliosis using three dimensional magnetic resonance imaging. AB - STUDY DESIGN: This report examines a technique for measurement of axial vertebral rotation using magnetic resonance imaging. OBJECTIVES: To assess the reproducibility of three-dimensional magnetic resonance imaging in the measurement of vertebral rotation at individual endplates in patients with adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Deformity in the sagittal and coronal planes in patients with adolescent idiopathic scoliosis can be readily assessed from plain radiographs, but the degree of deformity in the axial plane is more difficult to determine. Plain film techniques have inherent inaccuracies because of loss of definition of anatomic landmarks, and the use of computed tomography is limited by the high radiation dose associated. Magnetic resonance imaging provides a means of imaging scoliotic deformity that allows multiplanar reconstruction and that involves no use of ionizing radiation. METHODS: Ten patients with adolescent idiopathic scoliosis were imaged in a Siemens 1-Tesla impact scanner. Three-dimensional volume images of the apical five vertebrae were obtained in the axial plane and were postprocessed through multiplanar reconstruction. Sections through the superior and inferior endplates of each vertebra were selected in the sagittal and coronal planes, allowing axial reconstructions to be obtained in the plane of each endplate. Vertebral rotation was measured by identifying datum points on the inner surfaces and at the junction of the laminas and comparing the angle subtended by these points with a vertical drawn by the computer. Measurements were obtained from the single scanning sequence on two occasions by one observer and on one occasion by a second observer. Interobserver and intraobserver error was evaluated and correlation with readings obtained from plain films using Perdriolle's torsiometer method assessed. RESULTS: The interobserver variation had a mean of 3.02 degrees (range, 0-10 degrees) and a 95% confidence interval of [2.51 degrees, 3.53 degrees]. The intraobserver variation had a mean of 2.56 degrees (range, 0-7 degrees) and a 95% confidence interval of [1.83 degrees, 3.29 degrees]. The mean difference between measurements obtained from magnetic resonance imaging and plain film was 3.29 degrees (range, 0-12 degrees) with a 95% confidence interval of [1.43 degrees, 5.15 degrees]. CONCLUSIONS: The degree of vertebral rotation can be accurately and reproducibly assessed by three dimensional magnetic resonance imaging. Measurements can be made through individual endplates that allow assessment of the relative amount of intervertebral and intravertebral deformity. PMID- 9355224 TI - Measurement properties of the RM-18. A modified version of the Roland-Morris Disability Scale. AB - STUDY DESIGN: This investigation had two components: one was an item analysis that examined data obtained at the initial patient assessment, and the second was a validation study that used a pretest-posttest design. OBJECTIVES: The authors' goal, in this study, was to determine whether a shorter version of the Roland Morris Questionnaire could be developed with measurement properties equal to or better than the original 24-item questionnaire. SUMMARY OF BACKGROUND DATA: The measurement properties of the Roland-Morris Questionnaire have been shown to be better than or equal to competing measures. A number of studies have reported modified versions of the Roland-Morris without providing the measurement properties of the modified tool. METHODS: The item analysis investigated endorsement frequency, interitem correlations, item-corrected item total correlations, and coefficient alpha with various combinations of items deleted. The validation study examined reliability, concurrent validity, and longitudinal validity (sensitivity to change). The analyses included comparisons with the Oswestry and Jan van Breeman Pain Questionnaires. RESULTS: The item analysis suggested than six items could be detected from the Roland-Morris Questionnaire. The validation study demonstrated that the shorter version, named the RM-18, has measurement properties that are equal to those of the longer version. CONCLUSIONS: The RM-18 can be used as an outcome measure in clinical trials or as a tool to aid in decision making concerning individual patients. In either case, its measurement properties are equal to those of the 24-item Roland-Morris Questionnaire. PMID- 9355223 TI - Early childhood abuse in chronic spinal disorder patients. A major barrier to treatment success. AB - STUDY DESIGN: Prevalence rates of childhood abuse, socioeconomic outcome data, and levels of psychopathology were evaluated for graduates of a functional restoration program for chronically disabled spinal disorder patients in a workers' compensation environment. OBJECTIVES: To describe psychological profiles and evaluate treatment outcomes for chronic spinal disorder patients with a history of childhood abuse. SUMMARY OF BACKGROUND DATA: There is increasing evidence to indicate that traumatic childhood events may leave adult survivors psychologically distressed. It is possible that because of this level of psychological distress, chronic spinal disorder patients may be unable to return to a productive life-style after completing a rehabilitation program. METHODS: Two hundred ninety-nine male and 174 female patients from a cohort (N = 473) of consecutive graduates of a functional restoration program were assessed for the presence of childhood abuse by structured interview. Prevalence rates were compared with a comparison group of subjects without a history of chronic spinal disorder disability. In addition, the 79 chronic spinal disorder patients with a history of childhood abuse were compared on several socioeconomic outcomes with a matched group of workers with chronic spinal disorders without a history of childhood abuse. Psychopathology in the two groups of chronic spinal disorder patients was evaluated using Diagnostic and Statistical Manual of Mental Disorders criteria, the Minnesota Multiphasic Personality Inventory, and Symptom Checklist-90-Revised. RESULTS: A history of childhood abuse was found to be related to a higher level of psychological distress in chronic spinal disorder patients. In addition, poorer socioeconomic outcomes, such as lower work retention rates and higher postrehabilitation operations to the same area of injury, were found in the chronic spinal disorder patients with a history of childhood abuse compared with workers without a history of childhood abuse in whom chronic spinal disorders developed. CONCLUSIONS: These results demonstrate that although a history of childhood abuse is associated with greater psychosocial disturbances in chronically disabled spinal disorder patients, such disturbances do not interfere with an initial positive response to an effective tertiary rehabilitation program such as functional restoration. However, a history of childhood abuse may be related to poorer socioeconomic outcomes after discharge from rehabilitation programs. Additional treatment options may be needed for these patients. PMID- 9355225 TI - A comparison of surgeon's assessment to patient's self analysis (short form 36) after far lateral lumbar disc surgery. An outcome study. AB - STUDY DESIGN: Between 1984 and 1994, 170 patients had surgery for far lateral discs. Patients were assessed by the surgeon as having poor (no improvement, increased deficit), fair (mild improvement, moderate residual deficit), good (moderate improvement, mild residual deficit) or excellent (marked improvement, no deficit) physical outcomes. The Medical Outcome Trust's SF-36 survey was completed by 76 (45%) patients, using one interviewer. OBJECTIVES: Patient-based outcome studies are becoming increasingly important. A surgeon's assessment of outcome was compared with the patients' self assessment (Short Form 36) after far lateral lumbar disc surgery. SUMMARY OF BACKGROUND DATA: The SF-36 survey provides measures on eight dimensions: physical function, role physical, bodily pain, general health, vitality, social function, role-emotional, and mental health. METHODS: Patients averaged 60.1 years of age, and included 43 men and 33 women. Patients were last examined an average of 9.1 months after their surgery, and were interviewed by telephone an average of 2.8 years later. RESULTS: Patients completing the survey were evaluated on their last visit to the surgeon as having excellent (32 patients), good (24), fair (12), and poor (8) outcomes. Overall correlations between the surgeon's assessment and all 76 patients' SF-36 scores were modest. However, for those patients examined within 4.5 years of the surgeon's assessment (n = 56), correlations were statistically significant for 6 of the SF-36 measures. Only general health and social function showed correlations less than 0.25. CONCLUSIONS: The surgeon's assessment was a particularly good predictor of SF-36 measures if the surgeon assessed the patient within the past 4.5 years. The SF-36 should be useful for large-scale outcome studies. PMID- 9355226 TI - Anterior lumbar interbody fusion. A minimum 10-year follow-up. AB - STUDY DESIGN: One hundred eight patients from a consecutive series of 125 anterior lumbar interbody fusions were invited to take part in a clinical outcome assessment (including plain radiography and magnetic resonance imaging of the lumbosacral spine) more than 10 years after the original surgery. OBJECTIVES: By standardizing the reporting of outcome, to determine whether the duration of patient follow-up influences the outcome of surgery, with particular reference to the effects of compensation and psychological status. SUMMARY OF BACKGROUND DATA: The success rates of lumbar spinal fusion surgery reported in the literature vary widely. The lack of standardization of measures of patient outcome limits the value of study comparisons. Evaluation of the efficacy of spinal fusion is further compounded by the adverse effects of both compensation and psychological disturbance on the reporting of outcome. METHODS: One hundred three patients agreed to take part in a clinical outcome assessment by completing a comprehensive low-back questionnaire that included demographic, compensation, and employment details. Eighty-seven of these cases also agreed to undergo radiographic evaluation and magnetic resonance imaging of the lumbar spine. Subjective assessment of outcome was based on a 10-point analog pain scale as well as patient opinion regarding the success of surgery. A more objective assessment of outcome was made using the Low-Back Outcome Score. Psychological status was determined by combining the Modified Somatic Perception Questionnaire and the Zung Depression Scale. The effects of radiologic fusion, compensation status, psychological status, and reoperation on the various outcome measures were assessed and compared with the results reported in a separate but similar series of patients with a minimum follow-up of 2 years. RESULTS: Seventy-eight percent of patients rated themselves as having "complete relief" or "a good deal of relief," but only 34% fell into the "excellent" or "good" category using the Low-Back Outcome Score. The clinical outcome was not associated with the presence of radiologic fusion and was not influenced by the compensation status. Psychological disturbance at review and reoperation, however, did influence the reporting of outcome and were significantly correlated with the Low-Back Outcome Score. With the exception of the effects of compensation, these results were remarkably similar to the findings in the 2-year study. CONCLUSIONS: The findings of the study suggest that the assessment of outcome of lumbar interbody fusion is strongly compounded by the psychological make-up of the patient and that this effect is maintained in the long term. However, the negative effect of compensation observed at 2 years seems to dissipate with time and becomes insignificant at 10 years. PMID- 9355227 TI - Spinal deformity in myelodysplasia. Correction with posterior pedicle screw instrumentation. AB - STUDY DESIGN: A retrospective review of transpedicular instrumentation used in a series of 24 patients with myelodysplastic spinal deformities and deficient posterior elements. OBJECTIVE: To describe the usefulness and efficacy of these instruments in the treatment of complicated myelodysplastic spinal deformity. METHODS: The mean preoperative scoliosis was 75.7 degrees (range, 39-130 degrees) in the 22 patients with scoliotic deformities; 4 patients with thoracic hyperkyphoses averaged 70.5 degrees (range, 46-90 degrees) and 10 patients with lumbar kyphoses averaged 80.5 degrees (range, 42-120 degrees). The instrumentation extended to the sacrum in 4 patients and the pelvis in 9; 10 patients also underwent anterior release and fusion and 7 underwent concomitant spinal cord detethering. At an average follow-up of 4.0 years (2.0-7.7 years; one patient died at 8 months), all patients have fused (with the exception of two lumbosacral pseudarthroses). RESULTS: At last follow-up, deformity measured 32.1 degrees scoliosis (range, 6-85 degrees), 30.8 degrees thoracic kyphosis (range, 24-35 degrees), and 0.0 degree lumbar kyphosis (range, 35 degrees kyphosis to 29 degrees lordosis). Three patients lost some neurologic function after surgery; two recovered within 6 months and one has incomplete recovery. No ambulatory patient lost the ability to walk. Five patients required additional surgical procedures; in three cases, there was instrumentation breakage associated with pseudarthrosis or unfused spinal segments. CONCLUSIONS: Pedicle screw instrumentation is uniquely suited to the deficient myelodysplastic spine. Compared with historical control subjects, these devices have proven capable of significant correction of both scoliotic and kyphotic deformities. This instrumentation appears particularly useful in preserving lumbar lordosis in all patients and may preserve more lumbar motion in ambulatory myelodysplasia patients. PMID- 9355228 TI - Delayed infection after elective spinal instrumentation and fusion. A retrospective analysis of eight cases. AB - STUDY DESIGN: A retrospective analysis of eight cases of delayed spinal infection after elective posterior or combined anterior and posterior spinal instrumentation and fusion. OBJECTIVES: These cases are reviewed to identify risk factors for delayed spinal infection after elective instrumentation and to describe the treatment of this complication. SUMMARY OF BACKGROUND DATA: Delayed spinal infection after elective spinal instrumentation and fusion is uncommon. This diagnosis is frequently difficult. METHODS: Five cases seen in the senior author's practice and three referral cases are reviewed. RESULTS: Of these eight cases, the organisms were Staphylococcus epidermidis in six cases, Propionibacterium acnes in one cases, and in the final patient, all intraoperative cultures were negative. Clinical presentations were variable; however, all patients reported back pain. Seven patients had elevated erythrocyte sedimentation rates, averaging 57 mm/hour. Only two had elevated white blood cell counts. No distant foci of infection were identified in any patient. Five patients were found to have at least one pseudarthrosis. All patients were treated with debridement, instrumentation removal, and primary wound closure over drains followed by a minimum 6-week course of culture-directed postoperative antibiotics. At an average follow-up of 18 months, no patient has evidence of infection. CONCLUSIONS: The diagnosis of delayed infection after elective spinal instrumentation and fusion requires a high index of suspicion. These infections may have been caused by intraoperative inoculation. All patients were successfully treated with debridement, instrumentation removal, and culture directed postoperative antibiotics. PMID- 9355229 TI - Management of flatback and related kyphotic decompensation syndromes. AB - STUDY DESIGN: The authors, in this retrospective study, examined a group of patients with flatback syndrome and a related kyphotic decompensation syndrome. Results of nonrealignment treatment as well as revision surgery with sagittal realignment were reviewed. OBJECTIVES: To determine effectiveness of physical therapy and limited surgical (instrumentation removal) as well as major realignment surgical treatment in the sagittally malaligned spine. SUMMARY OF BACKGROUND DATA: Flatback is a sagittal plane deformity associated with distraction instrumentation for scoliosis correction. Kyphotic decompensation syndrome involves malaligned fusions from the sacrum for disease other than scoliosis. Several studies describe surgical realignment for flatback involving instrumentation systems no longer commonly applied. Guidelines for a systematic approach to flatback and kyphotic decompensation syndromes are lacking. METHODS: Forty-eight patients with flatback and kyphotic decompensation syndromes were reviewed. Treatment groups were defined by treatment approach and level of previous fusion. Effectiveness of treatment was reviewed in terms of radiographic sagittal alignment and self-reported pain. RESULTS: Twenty patients were treated without realignment revision surgery. Twenty-eight patients were treated with anterior and posterior osteotomies and realignment with instrumentation. For patients originally fused to the sacrum, realignment averaged 12 cm. Pain was reduced from 7 to 3 (10-point scale). In patients fused to L4 or L5, realignment averaged 7 cm. Pain was reduced from 6 to 2. Magnetic resonance imaging revealed viable caudal discs in four patients who were consequently spared extension of fusion to the sacrum. CONCLUSIONS: Treatment without realignment surgery demonstrated long-term success in 27% of cases. The latter all had two intact discs below the previous fusion and sagittal malalignment less than 4 cm. Realignment surgery effectively reduced pain in patients failing a conservative approach. PMID- 9355230 TI - Spondylolysis associated with Arnold-Chiari malformation and syringomyelia. A report of two cases. AB - STUDY DESIGN: This is a report of two cases. OBJECTIVE: To document the occurrence and association of spondylolysis and Arnold-Chiari malformation Type I. SUMMARY OF BACKGROUND DATA: The association of spinal dysraphism has been reported with Arnold-Chiari Type II, but not with Arnold-Chiari Type I. METHODS: The senior author was involved in the care of these patients. All medical records, laboratory and radiologic investigations, and related literature were reviewed. RESULTS: The presence of cephalic and caudal neuropore maldevelopment may be present in various combinations. The presence of spondylolysis, with or without spina bifida occulta, associated with Arnold-Chiari malformation type I and syringohydromyelia, is demonstrated. CONCLUSIONS: In some patients, the presence of spondylolysis may represent a congenital anomaly and may be associated with cephalic neuropore maldevelopment, such as cerebromedullary malformation syndrome (i.e., Arnold-Chiari malformation Type I). PMID- 9355231 TI - Role of electron microscopy in transplant renal pathology. AB - The crucial role that electron microscopy plays in diagnostic renal pathology is undisputed. By allowing recognition of findings not identifiable by light microscopic evaluation, electron microscopy has contributed significantly to the understanding of renal diseases and has proven to be of unquestionable value in many diagnostic situations. However, the percentage of cases in which electron microscopic examination adds important information that is either key for establishing or confirming a diagnosis or provides valuable data that influence patient's management remains controversial. This figure depends on the renal biopsy service that is surveyed, but it is reported that on the average ultrastructural evaluation is of value in approximately 30 to 45% of the cases. Correct interpretation of a renal biopsy depends on the ability to correlate light, immunofluorescence, and ultrastructural findings. In contrast, the role of electron microscopy in the examination of renal transplant specimens remains controversial. Many centers do not use routine electron microscopy to examine these specimens and insist that there are only a few specific indications that require ultrastructural evaluation. There is general agreement among renal pathologists that electron microscopy is of importance in the evaluation of renal specimens from patients with proteinuria to distinguish between transplant glomerulopathy, recurrent or de novo glomerulonephritis in order to correctly manage these patients and predict survival of the graft. The other possible indications are much more controversial. This paper summarizes and critically reviews the literature available on this subject and defines recommendations based on the information available at the current time. PMID- 9355232 TI - Medullary carcinoma of the breast: an ultrastructural morphometric study of nine cases. AB - Ultrastructural and morphometric features of 10 medullary carcinomas of the breast (MC) were investigated. Cases with a long follow-up were selected by applying stringent histologic criteria. All tumors had a homogeneous appearance by light microscopy. Under transmission electron microscopy, they showed occasional intracellular lumen formation or keratinization. In one tumor squamous differentiation was prominent and diffuse. Tumors with lymph node metastases possessed over 40% more desmosomes than nonmetastatic tumors. The number of cells with three or more nucleoli per nuclear section was significantly higher in metastatic than in nonmetastatic tumors (p = .02). Classic cases of MC of the breast display a relatively uniform appearance. However, subtle differences can be identified between metastatic and nonmetastatic tumors by ultrastructural morphometry. Although these differences are not associated with changes in the outcome of patients in this study, they seem to bear some relationship to the peculiar behavior of MC. PMID- 9355233 TI - Myelin modifications in 8 cases of peripheral neuropathy with Waldenstrom's macroglobulinemia and anti-MAG activity. AB - Characteristic myelin modifications in patients with IgM monoclonal gammopathy and anti-MAG activity have mainly been studied in cases of undetermined significance, but also exist in cases with indolent Waldenstrom's macroglobulinemia, i.e., when lymphoplasmocytic infiltration in bone marrow is 15% or more, without any visceral involvement. Since 1983, the authors have examined nerve biopsies from 8 cases with Waldenstrom's macroglobulinelia by direct immunofluorescence examination on frozen sections and ultrastructural examination. At direct immunofluorescence, fixation of anti-IgM serum on myelinated fibers was present in 7 cases. At ultrastructural examination, a widening of some myelin lamellae at the periphery of a few fibers was visible in 8 cases. A few fibers with hypermyelination were present in 5 cases. In 2 of these 5 cases widening of some myelin lamellae was present in numerous fibers, 88% in one of them. Frequently, there was a major widening of some myelin lamellae with dilated lamellae present in the inner part of the myelin sheath. Certain lamellae were more dilated, up to 50 nm. Occasionally, enlarged lamellae were not compacted with each other. The authors also examined nerve biopsies from 36 patients with IgM monoclonal gammopathy of undetermined significance and anti MAG activity, but found only one case with major widening of some myelin lamellae. Five other cases with major widening of some myelin lamellae, 4 Waldenstrom's macroglobulinemia and 1 IgM monoclonal gammopathy of undetermined significance, have been reported. Given that demyelinating neuropathies are far more numerous in cases with IgM monoclonal gammopathy of undetermined significance, it is likely that cases of indolent Waldenstrom's macroglobulinemia are prone to develop major myelin modifications, possibly due to another mechanism, added to the classic anti-MAG activity. PMID- 9355234 TI - Localization of CD44 at the invasive margin of glioblastomas by immunoelectron microscopy. AB - Glioblastoma multiforme is a highly invasive primary brain tumor, which is known to strongly express the CD44 cell adhesion receptor. A number of experimental studies suggest that the interaction of this receptor with extracellular matrix (ECM) proteins such as hyaluronic acid may in part mediate human glioma cell adhesion and invasion of brain tissue. Although the expression of CD44 and its spliced variants in brain tumors have been extensively studied, there have been no reports localizing its expression to the invasive margin of the tumor. The authors used immunoelectron microscopy to investigate the expression patterns of CD44 in an in vitro organotypic invasion assay. Tumor spheroids initiated from the U373 MG human glioblastoma line were confronted with fetal rat brain aggregates in a spheroid coculture system. The CD44 expression appeared at the interface between glioblastoma tumor spheroids and brain tissue, as well as in the spheroid itself. CD44 immunoreactivity was not detectable in mature 21-day fetal brain aggregates. The findings provide direct evidence that CD44 is expressed at the confrontational invasive border between glioblastomas and brain tissue, further supporting its role in glioma cell-ECM recognition and attachment. PMID- 9355235 TI - Immunoelectron microscopic localization of plasminogen activator inhibitor type 1 (PAI-1) in smooth muscle cells from morphologically normal and atherosclerotic human arteries. AB - Vascular wall fibrinolytic system proteins are believed to play a pivotal role in atherogenesis. Tissue-type plasminogen activator (t-PA) and urokinase plasminogen activator (u-PA) influence persistence of luminal thrombi and proteolysis of extracellular matrix, respectively. The major physiologic inhibitor of t-PA and u PA is plasminogen activator inhibitor type 1 (PAI-1). All three of these fibrinolytic system proteins have been detected in vascular endothelial cells, smooth muscle cells, and macrophages by light microscopic immunohistochemistry. This study was undertaken to delineate, by immunoelectron microscopy, the loci of PAI-1 in smooth muscle cells from intact morphologically normal and atherosclerotic human arteries as well as in isolated and cultured smooth muscle cells from arteries. In intact vessels, PAI-1 immunoreactivity was associated with contractile filaments in cells in both normal and atherosclerotic tissues. Lipid-laden smooth muscle cells in atherosclerotic vessels were mainly of the synthetic phenotype and displayed lesser amounts of PAI-1 associated with rough endoplasmic reticulum and contractile filaments. Isolated smooth muscle cells exhibited either a contractile or synthetic phenotype. In the cells with a contractile phenotype, PAI-1 was associated with the contractile elements, whereas in the cells with a synthetic phenotype, the PAI-1 was associated predominantly with elements of the endoplasmic reticulum. Because PAI-1 is associated predominantly with contractile filaments in smooth muscle cells, the net amount of immunodetectable PAI-1 appears to be greater in contractile compared with synthetic phenotype cells. PMID- 9355236 TI - Tenascin-C expression in ultrastructurally defined angiogenic and vasculogenic lesions. AB - Tenascin-C (TN) is an extracellular matrix glycoprotein expressed during embryogenesis. Its distribution is restricted in normal adult tissues and is upregulated in tumors and inflammatory conditions. Twenty-five specimens were studied, including 7 reactive vascular lesions (6 cases of granulation tissue and 1 case of bacillary angiomatosis), and 18 vascular tumors (6 angiosarcomas, 7 hemangioendotheliomas, and 5 AIDS-related nodular type Kaposi's sarcomas). Formalin fixed-paraffin-embedded tissues were stained with monoclonal antibody to TN (DAKO) and with MIB-1 (AMAC). Heterogeneous expression of TN immunoreactivity was seen in all cases, with a diffuse pattern in bacillary angiomatosis and most granulation tissue cases and a focal pattern in angiosarcoma and most hemangioendothelioma cases. Kaposi's sarcoma cases showed both a focal and diffuse pattern of distribution. In most cases proliferation indices (PI) did not correlate with TN expression. Electron microscopy demonstrated active angiogenesis in bacillary angiomatosis and granulation tissue and vasculogenesis in angiosarcoma and hemangioendothelioma. The study demonstrated positive TN expression in reactive lesions with angiogenesis (granulation tissue and bacillary angiomatosis) and neoplastic lesions showing vasculogenesis (angiosarcoma and hemangioendothelioma), although with a different pattern of distribution. These results suggest that TN might be an important extracellular matrix glycoprotein in angiogenesis and vasculogenesis. PMID- 9355237 TI - The ultrastructure of liposarcomas with attention to "dedifferentiation". AB - Liposarcomas are among the most common soft tissue sarcomas. It is recognized that dedifferentiation can occur within a well-differentiated liposarcoma, but there is limited information concerning the ultrastructure of the dedifferentiated cells. A series of 8 cases has been studied by light and electron microscopy and compared with well-differentiated, myxoid, and pleomorphic liposarcomas. No definite evidence of lipoblastic differentiation could be found in the dedifferentiated cases. The tumor cells resembled atypical cells in the well-differentiated liposarcomas, supporting the close relationship between these two types of tumors. However, since no conclusive line of differentiation could be found in the dedifferentiated cases, this study supports the contention that these neoplasms are undifferentiated counterparts of well differentiated liposarcomas. PMID- 9355238 TI - Skeinoid fibers in mesectodermal leiomyoma of the ciliary body. AB - Unlike smooth muscle elsewhere in the body, the smooth muscle of the iris and ciliary body is derived from neuroectoderm (mesectoderm). Leiomyomas that arise from the ciliary body, and therefore are of mesectodermal origin, may resemble spindle cell neurogenic tumors by light microscopy. They show positive immunostaining for smooth muscle actin but negative staining for neural markers. Ultrastructurally, the cells have the features of smooth muscle cells. The authors report a typical case of mesectodermal leiomyoma in a 47-year-old woman in which skeinoid fibers, considered to be an ultrastructural marker of neurogenic spindle cell tumors, were frequent together with other ultrastructural features often seen in neuroglial cell tumors. The findings indicate that mesectodermal leiomyoma is unique in its histogenesis as well as in its morphology. PMID- 9355239 TI - Amphicrine medullary carcinoma of the thyroid with luminal differentiation: report of an immunohistochemical and ultrastructural study. AB - A case of amphicrine medullary carcinoma of the thyroid is presented. The patient was an 18-year-old female with nonhereditary MEN IIb, submucosal neuromas in the oral cavity, and a thyroid tumor that metastasized to regional lymph nodes. Histologically the thyroid tumor was composed of polygonal cells arranged in a solid/trabecular pattern admixed with mucus-producing goblet cells and displaying focal cytoplasmic lumen formation. Immunohistochemical stains were positive for calcitonin, carcinoembryonic antigen, and chromogranin. Electron microscopy demonstrated C-cells containing neurosecretory granules as well as intestinal type microlumina. The presence of goblet cells and intestinal-type microlumina in medullary carcinoma of the thyroid is reminiscent of amphicrine tumors of the gastrointestinal tract and supports the hypothesis that the parafollicular C cells of the thyroid may be of endodermal derivation. PMID- 9355240 TI - Renal malakoplakia: report of a case with multifocal involvement. AB - The authors report the light microscopic and ultrastructural features in one case of malakoplakia involving the kidney, the urinary bladder, and the skin. The kidney was excised. Lesions of the urinary bladder and the skin regressed after topical treatment with cholinergic agonists and antimicrobial drugs. This case illustrates the pathogenesis of malakoplakia and the possibility that early lesions can be cured with medical therapy before extensive tissue destruction has taken place. PMID- 9355241 TI - Gangliocytic paraganglioma of the duodenum: report of a case with immunocytochemical and ultrastructural investigation. AB - A case of gangliocytic paraganglioma is reported in a 70-year-old female presenting as a polypoid tumor of the second portion of the duodenum. Immunohistochemical and ultrastructural features of the tumor indicate that gangliocytic paraganglioma of the duodenum (GPD) represents an unique tumor originating from the neuroectodermal derivative exhibiting dual phenotypic expression toward paraganglionic and epithelial (neuroendocrine) cells. These findings correlate well with the literature and it is further suggested that GPD belongs to the histopathologic spectrum of tumors derived from the neural crest. The findings offer a plausible explanation for histogenetic possibilities of the occurrence of pure epithelial tumors (i.e., carcinoid-like tumors) in the paraganglionic systems. The patient remains with no evidence of disease 4 years following a simple polypectomy. PMID- 9355242 TI - Case for the panel. Intranuclear Birbeck granules in Langerhans cell histiocytosis. PMID- 9355243 TI - Case for the Panel. Relation between Golgi complex and multivesicular body in malignant tumors. PMID- 9355244 TI - Lipofuscin-iron association in pigmentosis tubae. PMID- 9355245 TI - Presence of a unique parainfluenza virus 3 strain identified by RT-PCR in visna maedi virus infected sheep. AB - The presence in farm sheep of a paramyxovirus closely related to parainfluenza virus type 3 (Pi3) from humans or cattle was confirmed using RT-PCR on RNA samples from lung cells from slaughtered animals. Sequencing and restriction enzyme patterns of the amplified fragment of the F gene confirms the distinctness of the isolate, and suitable PCR primers allow specific detection of the ovine virus. A study of the incidence of ovine Pi3 in samples from sheep with or without distinctive histopathological signs of maedi shows that it is uncommon in aged sheep with overt lentiviral disease, but it occurs at moderate frequency in lambs and may, in the presence of visna-maedi virus, contribute to early lesion formation. PMID- 9355246 TI - Characterization of a putative receptor protein for bovine viral diarrhea virus. AB - In a previous communication, we reported a 50-kDa cell surface protein from Madin Darby bovine kidney (MDBK) cells as a putative receptor for bovine viral diarrhea virus (BVDV). The present study delineates further characterization of the receptor protein. Protease treatment of cultured MDBK cells adversely affected the receptor, thus abolishing the binding of anti-D89 (BVDV anti-idiotypes) to the cells. However, pretreatment of the cells with either phospholipases or glycosidases did not significantly change the anti-D89 binding to the cells. Additionally, pretreatment of cell monolayers with proteases decreased BVDV attachment and replication in the cells. These results suggested that the receptor for BVDV is a protein in nature, and glycosylation and phosphorylation may not play a direct role in BVDV attachment to cells. The BVDV receptor gradually regenerated on the cell surface after the protease-treated cells were cultured in normal growth medium. Regeneration of the BVDV receptor to a normal level took about 4 h as indicated by flow cytometric analysis and this process was inhibited in the presence of cycloheximide, a protein synthesis inhibitor. The 50-kDa receptor protein purified by electro-elution inhibited BVDV infection in a plaque reduction assay. It also inhibited anti-D89 binding to cells as analyzed by flow cytometry. These data demonstrated the nature of the 50-kDa protein as a specific receptor for BVDV. PMID- 9355248 TI - Effect of bovine herpes virus-1, bluetongue virus and akabane virus on the in vitro development of bovine embryos. AB - Bovine oocytes enclosed by follicular epithelial (FE) cells were exposed to bovine herpes virus-1 (BHV-1), bluetongue virus (BTV) and akabane virus (AV), matured in culture for 24 h and then in vitro fertilized. In the BHV-1-exposed group, BHV-1 was isolated from medium 3 days after in vitro fertilization at a high titer, and a severe cytopathic effect was observed in the FE cells. Oocytes in the BHV-1-exposed group developed to the 2-cell to 8-cell stage, but failed to develop into blastocysts. In the BTV-exposed group, BTV replication was observed in the FE cells during in vitro embryo development into blastocysts. In the AV exposed group, no distinct AV replication was detected in FE cells during in vitro embryo development. These results indicate that these viruses failed to infect zona-intact oocytes or embryos during in vitro maturation and culture. PMID- 9355247 TI - Application of recombinant bovine viral diarrhea virus proteins in the diagnosis of bovine viral diarrhea infection in cattle. AB - The National Animal Disease Laboratory (NADL) vaccine strain of bovine viral diarrhea virus (BVDV) genes for gp48 and p80 were expressed in Escherichia coli. The BVDV-NADL gene for gp62 was integrated into a baculovirus genome for expression in Spodoptera frugiperda (Sf-9) insect ovarian cells. The antigenicity of baculovirus expressed BVDV protein was detected by anti-BVDV specific antibodies in an enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescent assay (IFA) and radio-immunoprecipitation (RIP). The recombinant proteins isolated from bacteria showed antigenic properties when analyzed by ELISA and immunoblotting using BVDV antibodies. The recombinant proteins were then used in ELISA or IFA to detect BVDV infection by testing 54 independent bovine serum samples. The baculovirus-expressed BVDV protein was used as an ELISA and IFA antigen, and the bacteria-expressed proteins were used as ELISA antigens. BVDV-NADL-infected Madin-Darby bovine kidney (MDBK) cell monolayers served as a control antigen. Statistical analysis showed a high degree of correlation between the reactivity of recombinants and natural antigens in ELISA using bovine sera. The results of ELISA or IFA proved there is a high degree of correlation with the virus neutralization. In the comparative ELISA assays, the insect-cell-mediated expression revealed greater specificity and sensitivity than the bacterial expression or the natural BVDV antigens produced by cell cultures. PMID- 9355249 TI - Characterisation of Aujeszky's disease viruses isolated from domestic animals and from a wild boar (Sus scrofa) in Italy between 1972 and 1995. AB - Following the isolation of a group I Aujeszky's disease virus from a wild boar in 1993, an investigation was carried out on 30 Italian Aujeszky's disease viruses (ADV's), isolated over a 23-yr period in 12 Italian regions, by means of restriction endonuclease analysis. All strains isolated between 1972 and 1984 belong to group I. From 1984 onwards group II isolates (II and II intermediate) replace group I isolates. The isolation of a group I virus in 1993 from a wild boar supports the hypothesis that ADV's may persist for several years in wild boar populations and that wild boars should therefore be considered reservoirs of Aujeszky's disease virus. PMID- 9355250 TI - Differentiation of Mycobacterium paratuberculosis isolates by rDNA-spacer analysis and random amplified polymorphic DNA patterns. AB - The ability to distinguish between isolates of Mycobacterium paratuberculosis isolates was studied using two molecular techniques. Nucleotide sequence analysis of the rDNA-spacer and random amplified polymorphic DNA (RAPD) analysis using decamer primers with GC contents of 60 to 70% were evaluated on 16 isolates of M. paratuberculosis from cattle. The rDNA spacer analysis did not discriminate between isolates as it revealed an identical sequence for all 16 strains tested but it showed one common difference to the sequence previously described for M. paratuberculosis J2A. In the RAPD analysis, 14 of the 60 decamer primers used resulted in distinct amplification products for most of the isolates. For seven of the primers the size of the amplification products varied among strains thus allowing the specific identification of eight of the 16 isolates; of the remaining eight isolates five could each be differentiated from 14 other isolates, two from 13, and one from 12 isolates. Therefore, these data illustrate the possibility of using RAPD-analysis with certain primers for the differentiation of individual M. paratuberculosis isolates. PMID- 9355251 TI - Distribution of Salmonella contamination in ten animal feedmills. AB - Detailed sampling of spillage and dust from milling equipment was carried out in nine animal feedmills, three of which were sampled twice. The salmonella isolation rate ranged from 1.1% to 41.7% of the samples and the most contaminated mills were those where the inside of the cooling systems for pellet or mash had been colonised by salmonella. A wide range of salmonella serotypes were isolated which included Salmonella typhimurium and S. enteritidis. Limited sampling every two weeks for an 18-month period in another animal feedmill showed marked variation in the contamination rate of samples and range of salmonella serotypes found. Contamination of ingredient intake pits and outloading gantries for finished products by wild bird droppings containing salmonella was also found in four mills. PMID- 9355252 TI - Radioactive technetium-99m labelling of Salmonella abortusovis for the assessment of bacterial dissemination in sheep by in vivo imaging. AB - We report the development and validation of a 99mTc-labelling technique of bacteria, applied to Salmonella abortusovis. The radioactive labelling is obtained using a pre-tinning step of the cells followed by direct incubation of S. abortusovis suspension with 99mTc-pertechnetate. Several procedures with different amounts of stannous tin (SnF2 or SnCl2) were evaluated. The selected method, respectful of bacterial viability, provided a 30% labelling yield. Viability of 99mTc-labelled bacteria was assessed by flow cytometry using rhodamine 123 and was demonstrated to be unchanged, turbidimetric measurements showing only a slight increase in the growth rate for radiolabelled cells. Incubation of 99mTc-labelled S. abortusovis with pronase, saponine and urea demonstrated labelling stability and suggested an intra-cellular localization for 99mTc. A preliminary study was also conducted in sheep to evaluate the value of the imaging of radiolabelled S. abortusovis. Spatial and temporal patterns of their in vivo dissemination in the lymphatic system after a sub-cutaneous injection were compared with control lymphoscintigraphic agents. These imaging data supported the assumption that the radioactivity detected in vivo was proportional to the number of 99mTc-labelled bacteria. PMID- 9355253 TI - Immunological detection of sheep experimentally infected with strains of Mycobacterium avium subspecies containing insertion sequence IS901/IS902 and a 40 kDa protein. AB - A monoclonal antibody raised against a 40 kDa protein present in certain M. avium strains (IS901/IS902 positive) was used for developing a blocking ELISA. Sera from experimentally infected sheep were evaluated by indirect ELISA, AGID and blocking ELISA. The blocking assay proved to be highly specific for differentiation of sheep infected with different subspecies of M. avium. PMID- 9355254 TI - Development of an ELISA technique for serodiagnosis of bovine paratuberculosis. AB - A local clinical Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) isolate was cultivated in large-scale culture. A procedure for efficient preparation of a lipoarabinomannan-containing antigen was developed and standardized; 25 mg of purified antigen were obtained per gram of bacterial wet weight. An ELISA based on this antigen was developed. Intra- and interassay variation were determined to be 20% and 27%, respectively. The ELISA was evaluated using the sera of groups of 39 non-randomly selected and 92 randomly selected animals from which ileocaecal lymph nodes were cultured to isolate viable M. paratuberculosis. Combining the results of both groups a positive predictive value of 74% and a negative predictive value of 99% were calculated. The ELISA was licensed by the German regulatory agencies and, in a direct comparison, was found to be superior to both other licensed assays. PMID- 9355255 TI - Dermatophilus congolensis: strain differences in expression of phospholipase activities. AB - Interactions between Dermatophilus congolensis strains and with other bacteria of known haemolytic activities were used to elucidate the complex nature of haemolytic activities present in various D. congolensis strains. This was further analysed by measuring their specific phospholipase activities against defined substrates by thin layer chromatography. D. congolensis strains demonstrated haemolytic interactions (synergistic or antagonistic) with other D. congolensis strains and also other species of bacteria. Most isolates expressed lyso phospholipase-D activity, while various strains also expressed sphingomyelinase-D activity, phospholipase-A versus phosphatidylcholines and/or cephalins, phospholipase-D versus phosphatidylcholines or all these activities, under the culture conditions used. PMID- 9355256 TI - Seroepidemiology of canine leptospirosis on the island of Barbados. AB - Previous surveillance in Barbados documented the absence of infection with Leptospira serogroup Canicola in dogs. The aim of this study was to survey the current state of canine leptospirosis in Barbados, 10 years after the last survey. Sera from 78 unwanted dogs scheduled for euthanasia and 61 dogs suspected of having acute leptospirosis were tested by microscopic agglutination (MAT) and by an ELISA method adapted for canine IgM and IgG antibodies. The seroprevalence in unwanted dogs was 62% (48/78), at an MAT titre of > or = 100. The majority of animals had low titres, suggestive of previous infection. Serogroup Autumnalis was the most common reactor (45%), followed by serogroups Icterohaemorrhagiae and Australis (each 16%) and Pomona (13%). Serogroup Ballum was uncommon in this group. The seroprevalence determined by MAT in acutely-ill dogs was 75% (46/61). The most common predominant serogroup was Icterohaemorrhagiae (36%) followed by serogroup Australis (13%), while serogroups Autumnalis and Ballum were also of little significance. Paired specimens were available from eight acutely-ill dogs. One animal was seronegative while five dogs showed evidence of seroconversion. An IgM-ELISA titre of > or = 320 was used to confirm current infection in eight of these nine animals. Previous studies in Barbados showed a higher prevalence of serogroup Icterohaemorrhagiae than of Autumnalis, but the relative frequency of these two serogroups may be changing. The high seroprevalence in dogs is of public health concern because the close contact between dogs and man may provide the link between a reservoir in the environment and susceptible humans. PMID- 9355257 TI - Detection of Clostridium perfringens type D epsilon antitoxin in serum of goats by competitive and indirect ELISA. AB - Indirect and competitive ELISA techniques were developed and their ability to detect antibodies to Clostridium perfringens epsilon toxin in goat serum was compared. Different dilutions of a hyperimmune goat serum, in serum from a colostrum-deprived kid, were used as positive controls, while sera from eleven colostrum-deprived kids were used as negative controls. The epsilon toxin antibodies in the hyperimmune serum were also measured by mouse neutralisation test (MNT). The correlation coefficient between both the indirect ELISA technique and MNT was 0.99, while the same coefficient for the competitive ELISA was 0.98. Both the indirect and competitive ELISAs proved to be rapid, simple, sensitive and specific for detecting antibodies to C. perfringens epsilon toxin in serum of goats. PMID- 9355258 TI - Detection of Mycobacterium avium subsp. paratuberculosis in ovine tissues and blood by the polymerase chain reaction. AB - A direct polymerase chain reaction (PCR) test was applied to DNA extracted from blood, liver, ileocecal lymph node and ileum from twelve ewes in poor condition with histologically confirmed paratuberculosis and ten clinically normal sheep which had no evidence of paratuberculosis. The assay was compared with four serological tests: complement fixation test (CFT), gel diffusion test (AGID) and two enzyme-linked immunosorbent assays (ELISA). The PCR detection rate of Mycobacterium avium subsp. paratuberculosis, when results of single tests were interpreted in duplicate, was 72% for ileocecal lymph node, 90% for liver, and 100% for ileum in sheep with confirmed paratuberculosis. A single PCR test detected the target DNA in 66% of 0.5 ml blood samples. Sensitivities of serological tests compared with histological diagnosis were: 33% for CFT, 66% for AGID, 75% for the Central Animal Health Laboratory (CAHL) ELISA, and 83% for the 'modified' Commonwealth Serum Laboratories (CSL) ELISA. The PCR assay gave no positive reaction in samples collected from 10 sheep considered to be free of paratuberculosis. Similarly, all four serological tests were also 100% specific. The results raise some hope for the development of a PCR-based test using liver biopsy specimens, or possibly blood, in the diagnosis of paratuberculosis in sheep. PMID- 9355259 TI - Persistence in bovine mastitis of Staphylococcus aureus clones as assessed by random amplified polymorphic DNA analysis, ribotyping and biotyping. AB - Staphylococcus aureus isolates (N = 40) from bovine mastitis were characterized by random amplified polymorphic DNA-PCR (RAPD-PCR), ribotyping and biotyping. The isolates were collected in the veterinary surveillance area of the Ambulatory Clinic, Faculty of Veterinary Medicine, University of Helsinki from 20 quarters during the acute phase of infection and from the same quarters 3 weeks after cessation of therapy. The aim of the study was to compare the S. aureus isolates taken from the same quarter at different times to verify persistence of virulent strains in infected quarters and to compare the discriminatory power of the diagnostic methods. Using all methods (except for a commercial diagnostic test), the paired isolates of S. aureus were identical. Results suggest that the chronic nature of S. aureus infections was due to the persistence of the original infective strain. More laborious ribotyping and the more convenient RAPD-PCR method produced identical results. The molecular methods differentiated the 40 isolates into 6 distinct genotypes. Biotyping produced partially identical results to RAPD and ribotyping. A commercial diagnostic test system identified only 3 S. aureus biotypes. PMID- 9355260 TI - The differentiation of Brucella species by substrate specific tetrazolium reduction. AB - In the development of a tetrazolium reduction assay to replace substrate stimulated oxygen uptake for the identification of Brucella species, nine tetrazolium salts were evaluated. Only the more readily reduced compounds (MTT and INT) detected increased metabolic activity with the more fastidious, slow growing strains and with B. suis strains on L-arginine and DL-ornithine. The assay was optimised with MTT. MTT reduction profiles offered with the medium on which the cells were grown. Cells grown on TSA gave profiles more similar to the published respirometric results than those grown on SDA. The optimal substrate concentration was 0.84 g l-1 and prolonged (> 3 h) exposure to substrate was necessary before adding MTT. MTT concentration was not critical and the OD was proportional to the MTT concentration between 0.03 and > 0.5 g l-1. MTT reduction was linear for 60 min after its addition. The reaction was, therefore, stopped after 60 min by adding formaldehyde solution. The optimised assay was evaluated with 71 strains of Brucella, representing all the species and biovars of the genus. Each strain was assigned to its previously identified species and sub groups were defined. PMID- 9355261 TI - Diagnosis of canine brucellosis by detection of serum antibodies against an 18 kDa cytoplasmic protein of Brucella spp. AB - An antigenic capture ELISA was developed to measure serum antibodies against an 18 kDa cytoplasmic protein of Brucella. This assay was used to detect anti-18 kDa reactivity in sera from 30 dogs having confirmed or suspected brucellosis. Antibodies against the 18 kDa protein were found in 26 of them, which were also positive by the slide agglutination test (2ME-RSAT). The overall correlation (positive and negative results) between the ELISA and 2ME-RSAT tests was 93.3%. Additionally, these sera were assayed by an indirect ELISA using a whole extract of cytoplasmic proteins of B. abortus (LPS-free CYT). The results of both ELISAs were coincident in 28 of 30 (93.3%) dogs having confirmed or suspected brucellosis. When a serological follow-up was performed on some dogs having confirmed brucellosis, antibody titers measured by both ELISAs showed a parallel progression. On the other hand, the capture ELISA showed good specificity, since a positive result was obtained only in 2 of 103 sera from healthy dogs. These preliminary results show that the ELISA for detecting serum antibodies against the 18 kDa cytoplasmic protein of Brucella could be useful for the diagnosis of canine brucellosis. This study also shows that the results obtained with this single protein of Brucella are equivalent to those obtained with the whole extract of cytoplasmic proteins. PMID- 9355262 TI - Histological studies of bovine herpesvirus type 4 infection in non-ruminant species. AB - The pathology of bovine herpesvirus type 4 (BHV-4) infection was studied in cats, rabbits and guinea pigs. Twenty kittens, twenty-two rabbits and ten guinea pigs, some treated with glucocorticoid-were inoculated with a BHV-4 strain of feline origin, via various routes of inoculation (conjunctival, intranasal, peritoneal). Clinical signs were recorded. After euthanizing at different post inoculation days macro- and microscopic changes were observed by necropsy and in hematoxylin eosin stained histological sections. The presence of the virus in organs was detected by immunohistochemistry and a nested PCR assay. Inclusion bodies and monoclonal antibody-stained cells were found in the conjunctiva, trachea, lungs, spleen and lymph nodes. Most of the lesions were localized to the respiratory and the immune system. The macro- and microscopic lesions and clinical signs were more severe in kittens and guinea pigs. The histological data indicated that cats, especially kittens, were susceptible for BHV-4 and the infection was not confined to the urinary bladder. PMID- 9355263 TI - Identification of Clostridium chauvoei in cultures and clinical material from blackleg using PCR. AB - An identification system for Clostridium chauvoei, using PCR amplification of the 16S rRNA gene (rrs) with specific oligonucleotide primers and subsequent restriction digestion of the amplification product is described. The specific oligonucleotide primers were designed based on the rrs gene sequences of C. chauvoei by comparing it to the DNA sequences of the rrs genes of its most closely related species Clostridium septicum and Clostridium carnis. A subsequent restriction digestion of the 960 bp amplification product was used in order to unambiguously identify C. chauvoei. The developed identification system was evaluated on clinical material during a recent outbreak of blackleg in cattle. Thereby, C. chauvoei was identified as the etiologic agent of the outbreak either directly from clinical samples of muscle, liver, spleen and kidney or from primary cultures made with this material. A comparison of the newly developed method with standard diagnostic tools for C. chauvoei showed that it has advantages over the immunofluorescence and is, therefore, a useful option to it. Moreover, the assay is a valuable tool for the phylogenetic identification of C. chauvoei which can assist to substitute the fastidious traditional identification methods and replace laboratory animal testing currently used. PMID- 9355265 TI - "101" questions and answers about your AARN registration renewal. PMID- 9355266 TI - Registered nurses gather throughout Alberta to offer insight into continued competence monitoring. PMID- 9355264 TI - Delegation and supervision of client care: guidelines for registered nurses. Alberta Association of Registered Nurses June 1997. PMID- 9355267 TI - Injury prevention programs: do they really make a difference? AB - Brain and spinal cord injuries are a leading cause of death and disability for the youth of Canada (Damba et al, 1996). The costs of these injuries to the individual, the family, and society at large are immense. Most of these injuries are preventable (Tator et al, 1993). Over the last decade, a number of injury prevention programs have been developed to address the high incidence of traumatic central nervous system (CNS) injuries in young people. However, what remains unclear is how effective these programs are in terms of altering risk taking behaviours. The following paper/presentation will highlight three injury prevention programs currently offered in the Toronto area: The Party Program. The Heroes Program, and The Think First Program. Each program will be outlined in terms of historical development and infrastructure, content, setting, format, and intended audience. In addition, each program will be evaluated based on criteria established by the author. Measurement of outcomes will also be addressed. PMID- 9355268 TI - Protocol for intervention and treatment of alcohol withdrawal. AB - The incidence of alcohol dependence/abuse in patients of a general health care facility is 35-50%. The diagnosis and treatment of patients experiencing or at risk of alcohol withdrawal is problematic. The admitting diagnosis is usually another medical condition, illness or injury. Signs and symptoms of alcohol withdrawal is complicated by pre-existing conditions. In an attempt to improve the quality of care, decrease the length of stay of these patients, and decrease demands on nursing staff, a protocol for intervention and treatment of alcohol withdrawal was developed on the orthopedic ward of Royal University Hospital. The protocol enables each nurse to assess. Intervene and initiate the proper referrals. The recognized tool of assessment used to identify at risk patients is the CAGE questionnaire. The Clinical Institute WithDrawal Assessment for Alcohol scale is used to determine when it is appropriate to use Benzodiazepines. General nursing considerations are addressed through a pre-printed care plan. Nurses refer to social work, Alcoholics Anonymous and make use of available resource material. The protocol enables nurses to provide safe and effective care with few associated costs. Except for mass immunization, there is no other single intervention in health care that has the same far reaching consequences (Sullivan, 1995). PMID- 9355269 TI - Children's health resource centre. AB - The Children's Health Resource Centre was created to give children, youth and parents access to information concerning the health issues of children. As humans, whether we are children or adults, have a tendency to deal with situations better if we feel that we have some control. Knowledge empowers us gives us that control. It enhances our ability to cope and frequently improves our recovery as well as our recovery rate. The Resource Centre is there to supply that knowledge--to become a Provincial Inquiry line for children's health. This paper will encompass the why, how, when, response, effect and future plans of the Children's Health Resource Centre. PMID- 9355270 TI - The "nest"--neurological environment for safe therapeutics. AB - How to manage neurologically impaired patients is a challenge we all face. Lions Gate Hospital in North Vancouver, B.C. successfully developed a padded environment for confused, restless, and agitated patients. The following article describes how patients are managed without restraints in a safe environment. PMID- 9355271 TI - Epidurals under scrutiny in the United States. PMID- 9355272 TI - Episiotomy counts: trends and prevalence in Canada, 1981/1982 to 1993/1994. AB - BACKGROUND: The purpose of this study was to produce a minimum estimate of the prevalence of episiotomy use in Canada, and to investigate the trend in its use between 1981/1982 and 1993/1994. METHOD: A retrospective population case series study was conducted using hospital discharge abstracts. Outcome measures were the count of episiotomies performed during a 12-month period and the episiotomy rate per 100 vaginal births. RESULTS: For more than a decade, official statistics have significantly underreported episiotomy use by as much as 50 percent. In 1993/1994 at least 37.7 percent of women giving birth vaginally in Canada are known to have received an episiotomy. Between 1981/1982 and 1993/1994 its prevalence declined 29.1 percent, with the greatest decline occurring during the 1990s. This decline did not result from changes in parity in the population. The decrease in episiotomy use during this 13-year period is more than twice that found in the United States (a decline of only 13.6%). CONCLUSIONS: The reporting of official statistics on obstetric procedures in Canada should be modified to include all known cases of episiotomy. The observed downward trend in the rate of this procedure is encouraging, and is in the direction of evidence-based recommendations advocating its restrictive use. PMID- 9355273 TI - Reasons to stay, reasons to go: results of an Australian population-based survey. AB - BACKGROUND: Debate about early obstetric discharge is occurring simultaneously in several different countries. An Australian population-based survey of recent mothers investigated women's views about shorter postnatal stays and assessed the impact of early discharge on important maternal health outcomes. METHOD: Women's views and experiences of length of hospital stay were gathered by means of a statewide postal survey of all women who gave birth in Victoria, Australia, during two weeks in 1993. Questionnaires were mailed to women 6 to 7 months after the birth; 62.5 percent (n = 1336) responded. RESULTS: Most of the sample (64%) stayed in hospital for 5 or more days after the birth, 26.6 percent left on day 3 or 4, and 9.4 percent on day 1 or 2. Compared with women who stayed for 5 or more days, women who left in the first 48 hours were more likely to be multiparous; to have attended public models of care (public hospital clinic, shared care, public general practitioner, birth center); not to have private health insurance; to have experienced a lower level of obstetric intervention; and to have a very low income. Twenty-one percent of women who left within 48 hours, and 26 percent of those who left on day 3 or 4 described their stay as too short. Women who left on day 3 or 4, or within 48 hours, were not more likely to experience any of the adverse outcomes investigated in the study: breastfeeding problems, low confidence about caring for the baby, or depression 6 to 7 months after birth. Women who left on day 3 or 4 had slightly lower rates of breastfeeding at 3 and 6 months than women staying for longer or shorter periods. CONCLUSION: Concerns about possible adverse outcomes resulting from shorter postnatal stays were not borne out by the study findings. Large and carefully designed randomized trials are needed to resolve continuing uncertainties about the safety and possible benefits of shorter hospital stays. PMID- 9355274 TI - College students' knowledge and attitudes about cesarean birth. AB - BACKGROUND: Numerous clinicians and researchers have expressed concern about the necessity and potential adverse consequences of many cesarean births in the United States. The purpose of this study was to explore college students' attitudes and beliefs about cesarean section. METHODS: One hundred two college students (66% women) completed a 20-item questionnaire that asked if they viewed cesarean delivery as a potentially negative experience, as a normal or acceptable method of childbirth, and as medically necessary, and asked about their beliefs concerning risk and prevention of cesarean birth. RESULTS: The number of "undecided" responses in the study was striking (7.8% to 69.6% across the 20 items). In general, women and men responded similarly, although women were significantly more likely than men to say they would be profoundly disappointed if their babies had to be delivered by cesarean section. Despite expressing cynicism about the cesarean birth rate (40% agreed that many unnecessary cesarean births occurred) and not viewing the procedure as a normal way of giving birth (47%), most respondents (over 70%) disagreed that giving birth by cesarean would be a negative experience or would make a woman feel like a failure. CONCLUSION: A high level of uncertainty exists about certain aspects of cesarean birth among young women and men, highlighting the need for information for prospective parents. Most college students did not view the cesarean birth experience as either potentially negative or normal. Future research should explore coverage of cesarean birth in childbirth education classes and the roles physicians, nurses, and midwives play in preparing expectant parents for the possibility of cesarean delivery. PMID- 9355275 TI - Prenatal HIV screening in pregnant women: a medical-legal review. AB - Identifying pregnant women's human immunodeficiency virus (HIV) infection status provides them with the opportunity to seek appropriate treatment and to take measures to prevent vertical and horizontal transmission. Prenatal screening program options include targeting at-risk women, testing on a voluntary basis, or mandating prenatal HIV screening. When examining these options, the number of cases identified, programmatic costs, long-term health care costs, and legal implications must all be considered. Research indicates that targeting at-risk women misses a significant percentage of seropositive women, although programmatic costs may be lower. It is difficult to ascertain the difference between voluntary and mandatory programs with respect to the number of cases identified and treated. As a result, long-term savings are difficult to calculate. Mandatory programs would have the greatest direct costs and place the greatest burden on the woman's constitutional rights. By making HIV counseling and testing a routine component of prenatal care, voluntary programs could achieve the benefits of prenatal HIV screening without violating the woman's civil liberties. PMID- 9355276 TI - "I gotta push. Please let me push!" Social interactions during the change from first to second stage labor. AB - BACKGROUND: Forms of social interaction may occur among the participants in medicalized births, in which a woman in labor is experiencing strong involuntary urges to push but has not yet been found to have a completely dilated cervix. This article examines the social events and communications that occur at the change between first and second stages of labor. METHOD: Three cases are described from videotapes of women in the second stage of labor and their caregivers. RESULTS: Several social and interactive features occurred, in which (1) the caregiver, usually a nurse, by invoking the "no pushing rule," insisted that the woman suppress her involuntary urges to push; (2) both the caregiver and the parturient displayed an orientation toward the future and the eventual certification of full cervical dilation by a designated authority, usually a physician, regardless of the actual state of the woman's cervix or her involuntary urges to push; and (3) the certification process marked a ritual transition to "official" second stage labor, in which the woman's involuntary urges were considered appropriate and actively encouraged. CONCLUSION: A discrepancy between a laboring woman's sensations and caregivers' ideas about how labor should be conducted has implications for clinical care of women, wherein the goal should be to facilitate the woman's accomplishment rather than to direct the "delivery." PMID- 9355277 TI - Toward lower cesarean birth rates and effective care: five years' outcomes of joint private obstetric practice. AB - BACKGROUND: Many ways to improve perinatal outcomes, deliver cost-effective care, and increase client and caregiver satisfaction have been suggested. This article adds to the body of such literature by describing a joint practice in California and reporting five years of its outcomes. METHOD: Frequency data were recorded prospectively for all pregnant women seen between January 1, 1991, and December 31, 1995. Overall statistics and variable-specific frequencies were then analyzed for the 1303 consecutive singleton births that occurred during this period. RESULTS: The primary cesarean birth rate for the sample was 6.5 percent, the total rate was 9.1 percent, and the rate for women having their first full-term pregnancy was 11.3 percent. Of all women with a previous cesarean birth, 72.2 percent delivered vaginally. The success rate of attempted vaginal births after cesarean was 83.5 percent. Instrument deliveries were performed for 2.0 percent of births, and the frequency of third- or fourth-degree lacerations was 3.0 percent of all vaginal births. Transfers to a tertiary neonatal intensive care unit were 1.3 percent, and the perinatal mortality rate was 5.4 per 1000 births (corrected for serious anomalies: 3/1000). The preterm birth rate (including maternal transfers) was 2.0 percent. CONCLUSION: An obstetric practice in a private community hospital setting that effectively used obstetricians, nurse midwives, and nurse practitioners reported low rates of cesarean birth, preterm birth, severe lacerations, instrument deliveries, and legal incidents, and excellent cost-effective maternal and neonatal outcomes. PMID- 9355278 TI - Maternal health care at a Japanese American relocation camp, 1942-1945: a historical study. AB - BACKGROUND: From late summer of 1942 until the fall of 1945, approximately 120,000 ethnic Japanese were confined behind barbed wire within 10 relocation camps in the United States. Although histories have been written about the relocation camps, little data are available about women's lives. This study explored women's lives and experiences with pregnancy, childbirth, and child care in a Japanese-American relocation camp. METHODS: Twenty women who were ages 18 to 31 years at the time of internment at Heart Mountain, Wyoming Japanese American Relocation Camp, and one caucasian nurse who worked in the obstetric unit of the camp's hospital were interviewed. Archival, demographic, and historical data, including some prenatal records, provided information about maternity and public health care for pregnant women and new mothers. RESULTS: Obstetric hospital practices were typical of the 1940s in the United States. Community public health services for new mothers included formula kitchens and well-baby clinics. Infant mortality statistics from 1942 to 1945 at Heart Mountain were comparatively better for the same time period than for the state of Wyoming or the United States. These outcomes may have reflected the camp's extensive social and family support, adequate housing and food, and universal access to maternity services. CONCLUSIONS: The Heart Mountain internment provides a story about how women's lives are impacted by war. Since World War II, civilians, especially women and children, have increasingly been targeted during wars with profound impact upon the health of mothers and babies. PMID- 9355279 TI - Cesarean delivery rates in 1995 continue to decline in the United States. PMID- 9355280 TI - Sheila Kitzinger's letter from Europe: how can we help pregnant women and mothers in prison? PMID- 9355281 TI - Discharge packs and breastfeeding. PMID- 9355282 TI - Perspective: health care reform. PMID- 9355283 TI - Do nurses have the information resources and skills for research utilization? AB - While access to information resources and the skills to use them do not ensure that nurses will use nursing research in their practice, they are important facilitators. Mailed questionnaires to assess existing information resources, the information management skills of nurses, and what additional resources and training are required were returned by 67 of the 71 vice-presidents or directors of nursing in hospitals in two regions of Ontario. The two regions have similar information resources, nursing staff with research expertise, and opportunities for training in research and information management but there is variation among hospitals. Most vice-presidents agreed that nurses need better information resources and skills to access and evaluate professional literature. The rapidly developing field of information technology, including the Internet, provides potential for sharing resources and expertise. Nursing administrators can minimize barriers and help staff nurses recognize that information management skills enhance professional development and improve patient care. PMID- 9355284 TI - A survey of variables related to research utilization in nursing practice in the acute care setting. AB - Support from the work environment, attitude of nurses and availability of research findings are variables that influence the use of research in clinical practice. Acknowledging the importance of these variables in obtaining the goal of evidence based nursing practice, a nursing department of an acute care hospital determined that exploring these variables in its own setting was important to the successful planning of research education and utilization initiatives. This study describes the extent of selected factors (research support and availability, and attitude towards research) in relation to research utilization within this nursing department and based on the findings, suggests a number of ways nurse administrators can support research utilization, promote positive attitudes towards research and enhance availability of research findings in practice. PMID- 9355285 TI - Competencies related to medication administration and monitoring. AB - Reconfiguration of health service delivery has raised questions and concerns about the category of provider qualified to administer and monitor medications on inpatient units. The competency base (i.e. the scope of knowledge, skills and judgements) required, is dependent on the health status of the patient population and the medications prescribed. As part of a larger study addressing nurse competencies (Alcock, 1995), the required knowledge, skills and clinical judgement related to medication administration and monitoring for patients on two long term care units in a long term care facility and two tertiary care units in a general teaching hospital were determined. The total number of medications prescribed across the four units varied from 120 to 256 medications per unit. The number of different drug classes ranged from 27 to 32 per unit and the potential for drug-drug interactions ranged from 47% to 66% of prescribed medications across the units. The findings provide an information base for decisions related to education and to staffing. Various nursing delivery system options and factors which influence competency are discussed. PMID- 9355287 TI - Integrated delivery systems: the future for Canadian health care reform? AB - Influential health care leaders have recently said that it is a misnomer to refer to the current muddle of Canadian health care services and providers as a "system." Consequently, Canadian professionals and citizens have been exposed to increasing numbers of publications recommending that a move to fully and properly Integrated Delivery Systems (IDS) could rectify many of the difficulties with which our current non-system is faced. There is, however, a skepticism that the 'IDS movement' is just another in a series of fads that will come and go, without positively impacting the health care system. In this article, the authors provide arguments for why integration is critically required, an overview of types of integration, the qualities of a fully integrated system, and a discussion of the advantages and cautions to proceeding with IDS development. They conclude with the argument that--despite potential obstacles--the possible advantages of IDSs make it vital that we pursue integration. PMID- 9355286 TI - Implementing a hospital-wide pain management strategy. AB - The purpose of this article is to describe the process of undertaking a large scale nursing pain management initiative, that we entitled "Pain Month." Several educational resources and strategies were employed in an effort to increase nurse's knowledge on the subject of pain management. Pre and post patient satisfaction surveys were conducted to test the effect of the education on actual pain management. The results showed improvements in actual pain scores and in satisfaction with pain management; however, several more improvements were shown to be necessary to achieve excellent pain management for patients. Follow-up initiatives such as a pain resource nurse program, an interdisciplinary pain committee and continued, patient surveys have been implemented to address some of these existing issues. PMID- 9355288 TI - Globalization of information: new challenges and new opportunities. PMID- 9355289 TI - Research in nursing and cultural diversity: working with first nations peoples. AB - The current Canadian demographic profile indicates a society that is ethnically, culturally, and racially plural (Masi, 1993). Such diversity in our population is likely to increase with broad-based immigration, the implications of which have not been well addressed in nursing and health care. While nurses and other health care professionals must attend to understanding the cultural aspects of health and healing, there is little published valid and reliable research to assist with this, especially with respect to specific populations. Research approaches and designs must be culturally suitable to the specific populations, to generate valid knowledge about their culture and to develop theory, and to translate that into culturally suitable nursing and health care. In the past, many culturally diverse groups have been the subject of research that has been culturally inappropriate, patronizing, culturally threatening, and disempowering. This paper discusses critical ethnography as a culturally suitable research method and describes its application to studies involving First Nations (FN) peoples. Important issues in doing culturally suitable research, such as partnerships, ethical concerns, and ownership, are also discussed. PMID- 9355290 TI - Early infant crying: child and family follow-up at three years. AB - Children who cried excessively at six to eight weeks of age were re-examined at two to four years of age to determine the enduring effects of excessive crying ("colic") on behavioural development, parent-child interaction, and family functioning. The more crying in early infancy, the more family disruptions occurred three years later (r = .29). Analyses showed that early crying had little impact on the children's later behavioural development. No significant major lasting effects on the family related to the infant's early crying behaviour were found. Families with sufficient social and economic resources can be reassured that problems related to early infant crying can be ameliorated over time. PMID- 9355291 TI - Strategies to address the methodological challenges of client-satisfaction research in home care. AB - While there is an abundance of recent client satisfaction research, methodological difficulties continue. This paper addresses common methodological challenges in securing useful feedback from elderly clients receiving home-care services. The methodological challenges include socially desirable response sets (SDRS), fear of reprisal, acquiescent response sets (ARS), and negative or positive wording of items. These contribute to an inability to capture salient dimensions of satisfaction and dissatisfaction important to home-care clients. Several data-collection strategies are proposed: guided interactive interviews, story-telling, scenarios, and rating of the importance of the dimensions of satisfaction and dissatisfaction identified from interview data. Each strategy is discussed using illustrations from a study on elderly clients' satisfaction with home-care services. Nurses and other health-care providers require credible feedback about client satisfaction in order to develop policy and to provide more appropriate and effective services to home-care clients. PMID- 9355292 TI - Patterns of caregiving following the institutionalization of elderly husbands. AB - The purpose of this study was to examine the caregiving career of older women following the institutionalization of their husbands. Informed by the interpretive perspective in sociology and Hughes's (1971) concept of career, the study employed a longitudinal, prospective, and descriptive design and combined the quantitative and qualitative approaches. The data used in the analysis were drawn from a larger study designed to explore the transition to quasi-widowhood and wives' responses to their husbands' institutionalization. The caregiving career of wives was seen as a pattern of frequent visiting and increasing involvement in the provision of care. Over the nine-month period of the study, two caregiving patterns emerged that were distinguished by a variety of circumstances and interactions. Wives who relinquished aspects of caregiving were more likely to be caring for husbands who were, in large measure, cognitively impaired. These wives reported good morale, few symptoms of depression, change in marital closeness, and satisfaction with aspects of institutional care. Wives who continued to be heavily involved in caregiving were more likely to have husbands who were physically impaired. They had depression scores indicative of moderate to severe depression, reported no change in marital closeness, and were dissatisfied with aspects of institutional care. PMID- 9355294 TI - The National Forum on Health. PMID- 9355293 TI - Fraying connections of caring women: an exemplar of including difference in the development of explanatory frameworks. AB - While research exploring diverse groups enhances understanding of their unique perspectives and experiences, it also contributes to the exclusion of such groups from mainstream frameworks and solutions. The feminist grounded theory method allows for inclusion of marginalized groups through theoretical sensitivity to feminist theory and theoretical sampling. This paper demonstrates how this approach results in an explanatory framework that accounts for diverse realities in a study of women's caring. Fraying connections were identified as women's initial response to competing and changing caring demands. The range of dimensions and properties of fraying connections was identified through theoretical sampling guided by the emerging themes and theoretical sensitivity to issues of gender, culture, age, ability, class, and sexual orientation. PMID- 9355295 TI - A biblical perspective of ethics and justice in nursing. PMID- 9355296 TI - Justice as an ethic in nursing. PMID- 9355297 TI - My philosophy of professional nursing. PMID- 9355298 TI - My attitude--the key to fulfilling my God-given potential. PMID- 9355299 TI - Ethical dilemmas for aged care. PMID- 9355300 TI - Ethical issues and the Christian nurse. PMID- 9355302 TI - Giving yourself. Using personal resources. PMID- 9355301 TI - Brain dead--how do I cope? PMID- 9355303 TI - God's cares for the carers. PMID- 9355304 TI - Caring for the carers in the midst of low moral and nursing shortage. PMID- 9355305 TI - Counselling in nursing. PMID- 9355306 TI - God calls another nurse-specialist. PMID- 9355307 TI - Spiritual care in nursing practice. PMID- 9355309 TI - Caring: a Christian perspective. PMID- 9355308 TI - Care in action. PMID- 9355310 TI - My challenges as a Christian nurse teacher. PMID- 9355311 TI - Spiritual care in public health nursing. PMID- 9355312 TI - The rights of Christian women in nursing. AB - In this paper I have attempted to suggest that the Christian nurse who recognises the importance and significance of their personal characteristics as a woman and as a nurse is able to practice a model of care which enables rather than disables individuals and communities. A disabling model is one where we have a false perception of who we are and what our rights and responsibilities are. This approach perpetuates the passivity of women. Women have a unique view of the world and are able to contribute in a way that men are not able to do. We must not allow ourselves to be defined by men but in understanding ourselves we can fulfil our potential. This is not contrary to Christian teaching but rather corrects a cultural view which has been imposed on us about the nature of women in the profession and in the church. The essence of the Christian faith is that Christ died for all; without this belief there is no Christianity. Christ did not die for men alone who in turn represent women. In Christ there is no male or female Jew nor Greek, In Christianity there can be no sexism and no racism. We have rights as women and as Christians. Let us not neglect to use them. PMID- 9355313 TI - To be a Christian in nursing. PMID- 9355314 TI - Mary Seacole, the Florence Nightingale of Jamaica. PMID- 9355315 TI - Can Christianity & feminism agree? PMID- 9355316 TI - 'You can be a less effective nurse as a woman'! PMID- 9355317 TI - History in nursing: Pat Ashworth. PMID- 9355318 TI - "My nursing license is under investigation. What do I do"? PMID- 9355320 TI - Task force on workplace violence: Part II: Action plans. PMID- 9355319 TI - An overview of recent criminal consequences for nursing practice. PMID- 9355321 TI - Conflict resolution: a short description of the mediation process for health care professionals. PMID- 9355322 TI - Changes in the workplace: Colorado nurses report the affects. PMID- 9355323 TI - The occupational health nurse: "keeping business in good health". PMID- 9355324 TI - Latex allergy--an update. PMID- 9355325 TI - Advocacy, our most important role as nurses--a student perspective. PMID- 9355327 TI - The five hundred year flood. PMID- 9355326 TI - Colorado nurses aid flood victims. PMID- 9355328 TI - What nurses stand for. PMID- 9355329 TI - Special report: competence assurance for registered nurses. Ad Hoc Committee on Competence Assurance. PMID- 9355330 TI - Saskatchewan Heart Health Survey--Part I. PMID- 9355331 TI - The use of restraints in Saskatchewan long term care facilities. PMID- 9355332 TI - Trends in healing. PMID- 9355334 TI - Therapeutic touch: energy-based healing. PMID- 9355333 TI - A day with a healer. PMID- 9355335 TI - Mindfulness: treasuring the moments. PMID- 9355336 TI - The naturopath's perspective. Interview by Mary Jo Kreitzer. PMID- 9355337 TI - An honest mistake, a courageous resolution. PMID- 9355339 TI - Clinical trials for diet and hypolipidemic therapy have generally included few female subjects. PMID- 9355338 TI - Providing care for the dying: one answer, many questions. PMID- 9355340 TI - The features of the Critical Care Card. PMID- 9355341 TI - Nursing care of patients with acute myocardial infarction: results of a national survey. PMID- 9355342 TI - The radial artery: an exciting alternative conduit in coronary artery bypass surgery. PMID- 9355343 TI - The left anterior small thoracotomy technique: a new approach for coronary artery bypass grafting. PMID- 9355344 TI - Caring for patients with third-generation implantable cardioverter defibrillators: from decision to implant to patient's return home. PMID- 9355345 TI - Acute care nurse practitioner: a new career opportunity for critical care nurses. PMID- 9355346 TI - Assisted suicide, recent judicial decisions, and implications for critical care nurses. AB - The passage of the Oregon Death With Dignity Act on November 8, 1994, heralded a wake-up call for healthcare professionals. Oregon, the first state to systematically "ration care" was thought to be a fertile ground for testing new and, some say, radical concepts in healthcare and government. Although the act was not implemented because it was tied up in legal action until February 1997, the fact that more than 50% of the voters in Oregon voted for it mandates that healthcare providers listen to their patients. Patients want more control of their pain, the way they die, and the resources spent on their care in the final days of their lives. Thoughts of future suffering engender great fear on the part of healthcare consumers. Concern exists that physician-assisted suicide in the ICU will affect not only physicians but also nurses, pharmacists, respiratory therapists, and other clinicians as terminally ill patients make requests for assisted suicide while in the acute and critical care setting of the hospital. Critical care nurses must examine their value systems, review the Code for Nurses, and make their own decisions about participation in deliberately ending lives of patients. With the impending Supreme Court decision due in July 1997, the court may leave these issues to the individual states, opening the door for assisted suicide to occur throughout the United States. Therefore, the possibility will remain that critical care nurses may be put in positions in which physicians are providing assistance to patients who wish to commit suicide and are requesting nurses' assistance to do so. PMID- 9355347 TI - The ethical imperative to treat pain in infants: are we doing the best we can? PMID- 9355348 TI - Thrombolytic therapy in stroke management. Interview by Marilyn Petterson. PMID- 9355349 TI - Obtaining a pulmonary artery wedge pressure. PMID- 9355350 TI - Communication: something to point to. PMID- 9355351 TI - Designing a conscious sedation program. A collaborative approach. AB - St. Elizabeth's Medical Center identified a need to change its guidelines for the administration of IVCS. A task force consisting of physicians, nurses, pharmacists, and administrators was formed for this purpose. Goals for the guidelines were identified utilizing resources provided from the Board of Registration in Nursing and the Board of Registration in Medicine, as well as other professional organizations and local hospitals. A consensus was reached and a document generated that received approval by the Medical Center. Implementation went smoothly because of widespread support for the goals of the policy. Quality improvement studies have been performed and the results have been used to change practice. The process is currently ongoing. PMID- 9355352 TI - Developing a competency-based program for conscious sedation. AB - This article reviews the growth of IVCS usage from OR to bedside, including criteria set by professional organizations and regulatory agencies, and presents the development of a competency-based program for the IVCS monitor. The planning and implementation process for the IVCS competency program is discussed in terms of phases selection of IVCS monitoring staff, training of staff, competency verification, and competency maintenance. Because health care institutions must address the professional and regulatory standards of practice in IVCS, quality management activities are essential. PMID- 9355353 TI - A comprehensive review of sedative and analgesic agents. AB - The intent of this article is to be a comprehensive, but by no means exhaustive, review of some of the agents used for CS. The major classes and their principal uses are presented: benzodiazepines, for sedation-hypnosis, anxiolysis, and, in the case of midazolam, amnesia; and opiates, for analgesia and sedation. Also included are the miscellaneous items etomidate and propofol, for sedation hypnosis; ketamine, for sedation and analgesia; and the phenothiazines and butyrophenones. One should consider how the interactions between and among these agents can be used for the benefit of the patient undergoing CS, and also the danger in combining agents and the necessity of constant monitoring. The reporting of ADEs is a recurring theme, the value of which cannot be overemphasized in modern medical practice, not only to satisfy accreditation requirements but also to ensure patient safety and improve therapeutic choices of medications. PMID- 9355354 TI - Administering conscious sedation. Operational guidelines. AB - The nurse must be aware of her or his role and responsibilities when implementing IVCS guidelines. Nurses performing IVCS must be knowledgeable of state and institutional guidelines for IVCS, medications included in IVCS, and the assessment, monitoring, and documentation required in caring for the patient receiving IVCS. The process of IVCS may seem tedious, but if an institution has clearly defined expectations, as provided in the IVCS guidelines, the process is much more understandable and can be readily instituted. The patient undergoing IVCS deserves the highest quality care possible with the fewest complications, a situation that can be achieved with proper preparation and implementation of an IVCS program. PMID- 9355355 TI - Conscious sedation in cardiovascular procedures. AB - CS is commonly used in conjunction with CV procedures. Although CS provides a means to manage patient anxiety and discomfort, it also poses risks for the CV patient. The RN in the CV procedure lab plays a key role in the management of patients receiving CS. By formulating an individualized CS plan that integrates procedure risks, underlying disease processes, and medication effects, the RN minimizes patient risks and optimizes symptom management. PMID- 9355356 TI - Conscious sedation during electrophysiology testing and radiofrequency catheter ablation. PMID- 9355357 TI - Conscious sedation and implantable devices. Safe and effective sedation during pacemaker and implantable cardioverter defibrillator placement. AB - There is a paucity of research on the use of CS during ICD or pacemaker implantation. Pacemaker patients generally require small amounts of i.v. sedation, in conjunction with local anesthetic agents, to relieve incisional discomfort. Because of innovations in ICD technology, little difference now exists between ICD and pacemaker placement. Studies have reported on the use of deep sedation during ICD implantations, but only one study addressed CS in ICD placement. Whether deep levels of sedation are necessary for DFT testing is not apparent from these studies. Two studies report giving sedation at a level consistent with deep sedation, yet partial loss of protective reflexes was not reported in one of the studies. Further research is needed to explore the use of sedation in ICD implantation. It may be that deep sedation for patient comfort is necessary only during DFT testing or for tunneling of leads to an abdominal generator site, and that CS is more than appropriate for the implant portion of the procedure. Research may disclose that agents such as midazolam and, to some extent propofol, provide an amnesic effect that may mitigate the need for deep sedation during DFT testing or lead tunneling. Another benefit of agents and analgesics such as Fentanyl, is the relatively short duration of sedation, which minimizes recovery time. Patients recover quicker than with general anesthesia and can be discharged the same day in some procedures (e.g., generator replacement). Cost-benefit studies also are needed to compare the cost of anesthesiologist-delivered sedation with that of nurse-administered sedation. A comparison study could explore complication rates, patient comfort, and costs for the two sedation delivery methods. PMID- 9355359 TI - Conscious sedation in the endoscopy setting. AB - Endoscopy patients are adequately managed under CS and require attentive and skilled nurses to manage their care owing to the complexity of the procedures performed and the health status of this unique population. Endoscopy nurses need expert skills in assessing patient's respiratory and cardiac status to prevent complications and intervene successfully in emergency situations. Endoscopy personnel need to assess for and know how to treat complications that can arise from the procedure or from oversedation. The emergency endoscopy patient presents a challenge to the team and requires effective team coordination. PMID- 9355358 TI - Sedation and analgesia for oncological patients undergoing interventional radiologic procedures. AB - Patient-specific factors and knowledge of drug therapy direct CS and analgesia practices for oncology patients undergoing interventional radiologic techniques. Nurses caring for oncology patients in this setting must understand the effects of cancer on anatomic structures and physiologic functions. Variability in responses to pain and drug therapy observed among oncology patients necessitates the need for highly individualized plans of care that sometimes deviate from standard CS and analgesia procedures. Therefore, sedation and analgesia policies that apply to oncology patients must include flexible dose ranges for drugs such as opioids and benzodiazepines and guidelines for titrating these drugs to optimal effects. Although frequent assessments of pain using subjective reports remain the most valid indicators of adequate pain control, familiarity with radiologic interventions and responses of anatomic structures to various types of noxious stimuli allows the nurse to anticipate aspects of procedures known to be painful. Attention to symptom management both during and after the procedure is critical because patients with progressive cancer may present with significant physical and psychological alterations. In summary, CS and analgesia practices for oncology patients are based on sound pharmacologic principles and influenced by myriad factors that can be considered only in the context of the patient's own uniqueness and experiences. PMID- 9355360 TI - Conscious sedation in the ambulatory setting. AB - Conscious sedation has become an important component of practice for many ambulatory procedures. Important factors to maintain the safety of this technique are proper patient selection, slow titration of the medication, continuous patient monitoring, proper education of the individuals administering the medication and monitoring the patient, and appropriate policies and guidelines in place in the ambulatory setting. When patient safety is the number one priority, conscious sedation can be a very effective, practical, and safe technique used in the ambulatory setting. PMID- 9355361 TI - Practical considerations in sedating the elderly. AB - CS can safely be administered to the elderly population. The definition of elderly is a subjective term. That is, each patient should be considered as an individual person with different physiologic and psychologic needs. Special attention must be given to ensure a safe environment for induction of sedation. The patient should be fully assessed, keeping in mind the physiologic changes that accompany aging. Elderly patients have an increased variability of drug response. Elderly patients have a decreased requirement for most anesthetic drugs. Elderly patients have an increased redosing interval. Continuous monitoring for signs of intolerance and cautious administration of sedation will help reduce the risks associated with sedation in the elderly. PMID- 9355362 TI - Conscious sedation in the pediatric population. Special considerations. AB - The child requiring sedation has unique needs. The nurse caring for pediatric patients must have adequate knowledge to incorporate the physical, emotional, and psychological differences between children and adults into the child's overall plan of care. Because of these differences, sedation of the child presents a challenge. The nurse must continue to assess his or her knowledge of all facets of sedative agents and monitoring principles in the pediatric population to provide safe, effective quality care to children and their families. PMID- 9355363 TI - Sedation in the critically ill patient. AB - The need for sedative therapy in the critical care setting is well established. It is an absolute therapeutic adjunct useful in decreasing both patient morbidity and mortality. Sedative endpoints, monitoring requirements, and recommended agents and dosages associated with sedation, however, remain poorly defined. Standardization is the key to optimal patient outcome at minimal cost. This article presents an overview of current trends and recommendations. Continuing research into these areas is strongly recommended to ensure consistent and safe practice. PMID- 9355364 TI - A nursing guide to conscious sedation. Clarification of current practice issues. AB - The practice of sedating patients for painful or constraining procedures has a long history and will continue to expand in the future. National standards guide the clinical management of all patients to achieve a standard of care for every CS experience. The JCAHO standards and professional organizational guidelines regarding CS should be reviewed. Development of comprehensive policy through a collaborative team approach is recommended. PMID- 9355367 TI - A journey into collaborative case management. PMID- 9355366 TI - Risk management considerations in conscious sedation. AB - Sedation care is an expanded nursing role that may have potential liability associated with it. Liability reduction requires consistent application of the principles of the nursing process. Properly prepared nurses with commitment to patient safety will discover that the art and science of nursing combine in this challenging area of patient care. PMID- 9355365 TI - The cost of administering intravenous conscious sedation. AB - CS is an alternative to general anesthesia or MAC in some diagnostic and therapeutic procedures and surgeries, and it can be equally effective in ensuring adequate sedation to facilitate completion of the procedure. It does come with some expense, although still not as costly as general anesthesia. Use of this technique must be undertaken with caution and concern for patient safety, however, and thus some additional expenses will be incurred. With careful selection of the agents used, proper equipment for monitoring, and extensive physician and staff education, the technique has the advantage over general anesthesia in terms of cost not only to the institution but also to patients and to third-party payers. PMID- 9355368 TI - A summary of the certification examinations from 1994 and 1995. PMID- 9355369 TI - Protein, diabetes, and nephropathy. AB - Evidence suggests that a low-protein diet (0.8 g/kg) is beneficial in persons with diabetes with the onset of macroalbuminuria. Despite problems with existing studies, preliminary evidence suggests that vegetable proteins are not detrimental to renal function and may be used to supplement or replace animal proteins. PMID- 9355370 TI - Computer-assisted diabetes nutrition education increases knowledge and self efficacy of medical students. AB - Medical students and physicians need to improve their understanding of the role of nutrition and the multidisciplinary team in diabetes care. To assist in this learning, an interactive computer program was developed that focused on prescribing diets for patients with diabetes. Parallel 10-item knowledge tests and an 8-item self-efficacy scale were used to evaluate the efficacy of the computer program among 41 third-year medical students. Mean knowledge scores increased significantly after using the computer program. Posttest knowledge scores for the medical students approached the level achieved by general practice dietitians with no diabetes specialty training. Mean self-efficacy scores increased significantly. The mean time spent on the educational component of the program was under 30 minutes. Computer-assisted diabetes nutrition education proved to be an efficient and effective method for teaching basic nutrition competencies to medical students. This program is available on the World Wide Web (http:/(/)medicine.aecom.yu.edu/diabetes/DEC.htm ) and may be a useful means for providing basic diabetes nutrition education to primary healthcare providers from a variety of disciplines as well as for medical students. PMID- 9355371 TI - Personal illness models of diabetes: parents of preadolescents and adolescents. AB - The purpose of this research was to explore personal illness models of parents of preadolescents and adolescents regarding diabetes mellitus. Personal illness models were defined as the parents' cognitive representations of the disease. Fifty-five parents of children ages 10 to 17 years with a diagnosis of insulin dependent diabetes mellitus were interviewed using a semistructured questionnaire. Data were content analyzed for common themes. Parents attributed the cause of diabetes to genetics coupled with a viral infection. Most believed the diabetes would last a lifetime but they were hopeful for a cure. Parents requested ongoing education for their children, support groups, counseling, one consistent healthcare provider, and intensive insulin therapy. Parents reported that the major problems caused by diabetes were increased structure of daily routines and that their children with diabetes felt different from healthy peers. Parents' fears about diabetes included long-term complications, early death, and severe insulin reactions. PMID- 9355372 TI - College students with diabetes: using focus groups and interviews to determine psychosocial issues and barriers to control. AB - College students with diabetes are at risk for improvised diabetes care due to their age, newly acquired independence, and erratic schedules. The purpose of this study was to employ focus groups and interviews to identity factors that affect the ability of these students to engage in appropriate self-care behaviors. Focus group and interview questions were developed to address variables of the Expanded Health Belief Model. Two focus groups and fifteen interviews were conducted. Barriers to successful diabetes management were time management, stress hypoglycemic reactions, diet management constraints, and inadequate finances. Several psychosocial issues that affected successful management also were identified. These issues were grouped into three categories: (1) inconveniences of diabetes management, (2) motivators to managing diabetes, and (3) social support issues. The findings show the value of formative evaluation that can then be used to design diabetes education programs to meet clients' perceived needs. PMID- 9355374 TI - Chronic care: a need in search of a system. AB - Currently, the bulk of our healthcare dollars is being spent on chronic illnesses and their associated complications. Given this use of resources, it is essential to identify, develop, and implement a system of care that is unique to those with long-term health concerns. Effective management of these problems depends largely on patient participation and strategies that are appropriate for the primary delivery site, which is the home. Such concepts are foreign to most current disease-management programs, and years will be required for acceptance and implementation. Quality long-term management requires a systematic, comprehensive system of care that is unique to the needs of both patients and providers. Such a system consists of a series of integrated, interdependent components that will be presented in this article. PMID- 9355373 TI - Diabetes in urban African Americans: functional health literacy of municipal hospital outpatients with diabetes. AB - Functional health literacy was assessed in 63 patients from the diabetes outpatient clinic, 20 from the general medicine clinic, and a total of 48 from two satellite medical clinics. All patients received a demographic questionnaire, visual screening, and the Test of Functional Health Literacy in Adults, an instrument with good validity and internal consistency used to measure the ability to read and understand medical instructions. Functional health literacy was adequate in only 47% of new patients at the diabetes clinic and only 25% of established patients at all sites. There were no significant differences in functional health literacy among established patients across all sites. Overall, patients' mean functional health literacy level was inadequate to marginal. Of the patients with inadequate functional health literacy, 43% denied difficulty in reading. Patient education strategies and materials are needed to address this important barrier to healthcare delivery. PMID- 9355375 TI - Feasibility of a kiosk-based patient education system in a busy outpatient clinic setting. PMID- 9355376 TI - Nurses indicted ... a wave of the future? PMID- 9355377 TI - [Model development in nursing science; the construction of a theoretical model]. AB - The aim of the paper is to describe the development of nursing theory. As an example, a theoretical model of the compliance of young diabetics and related factors is built. The content of the theoretical model is not described but the process of developing the model is presented. In the first phase, a hypothetical model of young diabetics' compliance was developed inductively. The data were collected by interviewing 51 young diabetics aged 13-17 years, by observing the behaviour of 18 of these young people during an adaptation course and by analysing their drawings (N = 17). The data were analysed by using continuous comparative analyses and content analyses. In the second phase, an instrument for testing the model was developed. For this purpose, data were collected with a questionnaire from young diabetics aged 12-17 years (N = 91). The content validity, construct validity and reliability of the instrument turned out to be quite good. In the third phase, the hypothetical model was tested and developed further by using LISREL (linear structural relations) analyses. The data were collected with a questionnaire from young diabetics aged 13-17 years (N = 346). In the fourth phase, the model was expanded using the qualitative data (N = 51). The categories which had been discovered by continuous comparative analyses were quantified. The data were analysed by cross-tabulation, the chi square test and discriminant analyses. A summary of the results has been presented by building a theoretical model of young diabetics' compliance and related factors. PMID- 9355378 TI - [Fathers' childbirth experience and nursing interventions]. AB - The purpose of the study was to describe the childbirth experience of fathers and nursing interventions used during labour and delivery. The data were collected by questionnaires from fathers attending delivery room during one month (n = 137). Five fathers refused to participate. The return rate was 81% out of 132 fathers. Data analysis included cross tabulations, factor analysis as well as content analysis for open ended questions. Nursing interventions which fathers found supportive were infant care, getting information and attention from the staff. Most of the respondents felt they got encouragement to take care of the infant. They also got information of the delivery and newborn's welfare. However, about half felt they got little encouragement to show their own feelings. Fathers found that being present at the delivery was an important part of their fatherhood. The best moments were attached to enjoyment of taking care of the newborn. Fathers felt themselves unable to help their partner enough. They felt very difficult to see their partner in pain. Most of the father were quite satisfied with nursing interventions used during labour. They wanted more attention to be paid to pain management. Fathers would like to get more advice during labour. PMID- 9355379 TI - [Use of qualitative diaries in health-related research]. AB - This article focuses on factors which are relevant while using a diary in qualitative health related research. Diary is a data collection method which has several advantages: it approximates the real activities of research persons, it diminishes memory errors which may occur in retrospective interviews, and it helps to better understand the lifeworld of the research persons. Diaries can be used as an independent data collection method or combined with other methods, such as semi-structured or deep-interviews or observations. Using a qualitative diary needs a careful consideration of the design and aims of the research in question. For these reasons water-proof instructions of using diaries are not eligible. Rather, the method should be applied each time it is used. In the area of nursing science there are several possibilities for applications. For example, the view-points of a hospital patient or a client of health care services can be reached and understood better. PMID- 9355380 TI - [Proceedings of the 1st representatives' assembly of the 25th Congress NAROC (Nurses' Association of the Republic of China)]. PMID- 9355381 TI - [Nosocomial infection control and reverse isolation]. PMID- 9355382 TI - [Instruction of home-based exercise for cardiac patients]. PMID- 9355383 TI - [Diabetes and exercise]. PMID- 9355384 TI - [Pain management: nursing perspective]. PMID- 9355385 TI - [The meaning of a good death for terminally ill cancer patients in Taiwan]. PMID- 9355386 TI - [Applying Peplau's theory in improving drug compliance of a schizophrenic patient]. AB - The purpose of this paper is to report the application of Peplau's theory to helping a schizophrenic patient deal with drug noncompliance. It was found that the main reasons for noncompliance were poor insight and lack of illness and drug related knowledge. Based on Peplau's theory of establishing a therapeutic relationship, giving guidance in medication and monitoring the effects of medication in order to improve drug compliance, the most important role of the nurse is as an assessor and educator. Nurse and patient were able to learn to achieve the common goal of drug compliance through the interactive human relationship. PMID- 9355387 TI - [Body image concerns and behavioral responses in a patient with cerebellopontine angle tumor]. AB - This case report investigates the behavioral responses of body image change of a cerebellopontine angle tumor patient. Twelve process recordings obtained from a field study comprised the data for analysis. The dimensions of the subject's body image change were body structure, body function, body sensation, and social function change. Two types of behavioral response were delineated and categorized as the following: (1) Identifying the individual's body image change, such as seeking information, seeking reassurance, and comparative behavior. (2) Maintaining the integrity of individual's body image, such as rationalizing behavior, maintenance behavior, cooperative behavior and seeking religious support. This paper reports how the investigators used nursing interventions to help the subject gradually get through the process and adapt to the body image changes properly. PMID- 9355388 TI - [Attachment process between a mother and a premature baby]. PMID- 9355389 TI - [Nursing management of problematic behavior in elderly dementia patient]. PMID- 9355390 TI - [Nursing care in renal transplantation]. PMID- 9355392 TI - [A report about palliative care in Japan]. PMID- 9355391 TI - [Uncertainty: a concept analysis]. PMID- 9355393 TI - [The league of nursing students in junior college]. PMID- 9355394 TI - [Report on and analysis of the first credential examination on critical care nursing]. PMID- 9355395 TI - [The nurse as HIV educator: tips for reaching the general public]. PMID- 9355396 TI - [A community-based approach to HIV/AIDS prevention for adolescents: a time for action]. PMID- 9355397 TI - [Concerns and nursing care of a family living with AIDS]. AB - AIDS is not solely a medical issue but also has profound implications for family relationships. This case report using the concepts of family nursing and case management presents the concerns of a family living with AIDS. The data was collected through continuous contacts with a patient during December 1994 to August 1996. This report's findings indicate that the concerns of the family living with AIDS fall into seven categories: The reasons for HIV infection, how to meet the patient's sexual needs, suffering during health care seeking, the effectiveness and side-effects of anti-viral drugs, home care skills, the light of hope, death issues. Professional nurses should provide care to both the AIDS patient and the family members. Using social resources and cooperating with other health professionals can promote the quality of life for families living with AIDS. PMID- 9355398 TI - [Medical precaution in human immunodeficiency virus infection]. PMID- 9355399 TI - [An exploration of sexual knowledge, attitudes and behavior in aboriginal elementary school students in the Ping-Tung area]. AB - The purpose of this study was to investigate sexual knowledge, attitudes and behavior of fifth and sixth grade students in aboriginal elementary schools in the Ping-Tung area. A structured questionnaire was administered to 1091 students who were selected by cluster sampling. The results showed: (1) The sexual knowledge score was low but sexual attitudes showed a positive trend. (2) 64.7% and 67.4% of students had at some time seen pictures of male or female sexual organs. (3) About 61% of students had seen sexual magazines or videotapes. (4) 66.2% of male and 88.1% of female students had heard about wet dreams or menstruation before their first experience; more than half of the students thought that wet dreams need treatment. (5) 17.8% of students had masturbation experience, and after that 59.3% of students had fear or guilt feeling. (6) Female students had significantly higher knowledge and attitude scores than male students, Demographic variables produced no significant difference in the above scores. (7) 42.4% of students most desired to know what phenomena indicate sexual maturity. (8) Sex knowledge had significantly positive correlation with sex attitude. PMID- 9355400 TI - [The application of Roger's science of unitary human beings for an adolescent with mental illness]. AB - The purpose of this paper is to report the application of Rogers' science of unitary human beings theory to exploring the influence of the environment and the nursing process on an adolescent with behavioral disturbance at a children's mental health outpatient. Rogers' theory is that the principles of resonancy, helicy and integrality explain the life process of human beings, and predict the development of the life process. In the paper, these principles were adopted throughout the nursing process to explore their relationship. We found that the nurse made use of varied nursing interventions, and that the interventions were influential in three aspects. In the aspect of integrality they could promote the interaction of client and environment energy field. In the aspect of helicy, they were able to reduce the complexity of the behavioral deviation. In the aspect of resonancy, they were able to slow down the evolution of acceleration of energy field between client and environment. PMID- 9355401 TI - [Nursing care in renal transplant patients with infection problems]. PMID- 9355402 TI - [Nursing care of a patient with pemphigus]. PMID- 9355403 TI - [Time concept and nursing]. PMID- 9355404 TI - [An exploration of the stressors in heart transplant recipients]. PMID- 9355405 TI - [Andragogy and health education among the elderly]. PMID- 9355406 TI - [Respiratory rehabilitation in patients with chronic obstructive pulmonary disease]. PMID- 9355407 TI - [Teaching strategies for the elderly]. PMID- 9355408 TI - [Palliative care of dyspnea in terminal cancer patients]. PMID- 9355409 TI - The OIIQ sets up a Youth Committee "Generation X" springs into action. PMID- 9355411 TI - [Reducing the risk of sudden death syndrome in infants]. PMID- 9355410 TI - [Knowing more about "capitation"]. PMID- 9355412 TI - [Test your knowledge--caring for a client undergoing surgery]. PMID- 9355413 TI - [The nurse and the Internet--foolish or a bright idea?]. PMID- 9355414 TI - [L'infirmiere du Quebec has navigated for you]. PMID- 9355415 TI - [The stakes of ambulatory care trends in mental health]. PMID- 9355416 TI - [Allergic? Don't panic!]. PMID- 9355417 TI - [Losing one's father at 20 years of age]. AB - At first intending to describe and understand how all family members experience mourning, three researchers at the Universite du Quebec a Trois-Rivieres found that very few studies have dealt with young men's reactions to the deaths of their fathers. The researchers then conducted a qualitative study, using a phenomenological approach, of this as yet little-explored subject. This study, Le deuil tel qu'experimente par les fils lors du deces de leur pere (Grieving as experienced by sons on their fathers' death), enabled them to describe and better understand what young men experience on their fathers' death. The father was an important figure in the subjects' lives, and his death was a pivotal experience for each of them. The family structure was shattered and, while each subject's experience was unique, their intense reactions resemble those described by grief theorists. In a detailed discussion of the results of the study, the authors relate these results to theories on grief and youth, and suggest applications in nursing practice. PMID- 9355418 TI - [Luc Poirier, nurse and entrepeneur. Interview by Sylvie Vallieres]. PMID- 9355419 TI - An interview with nursing graduates. Interview by Rivka Guttman. PMID- 9355420 TI - Nursing graduate employment: a survey of Anglophone CEGEPs in Montreal. PMID- 9355421 TI - Sharing the health challenge in critical times. PMID- 9355422 TI - Adverse reactions to the withdrawal of opioids and benzodiazepines in paediatric intensive care. AB - The aim of this study was to examine adverse reactions to the withdrawal of opioids and benzodiazepines among critically ill children. Although withdrawal reactions have been well documented in relation to substance abusers and their newborn infants, there has been little study of this phenomenon as an iatrogenic problem. We developed a graphical case study method for examining patterns over time, and applied this to five cases referred to us by the nursing staff of a 10 bed paediatric intensive care unit. A striking pattern of behavioural distress was clearly associated with the diminution of opioids and benzodiazepines. These adverse reactions were characterized by various combinations of inconsolable crying, tremors, jitteriness, irritability, gagging, vomiting, and feeding problems. These signs appeared as early as 1 h and as late as 24 h following a significant reduction in opioid and benzodiazepine infusion rates, sometimes following very short-term therapy. We elaborate an interpretation of this distress, in light of the multiple disruptions undergone by critically ill children, and conclude by outlining our recommendations for preventing/minimizing these adverse reactions. PMID- 9355423 TI - Reconnecting: the experiences of nurses caring for hopelessly ill patients in intensive care. AB - The aim of this grounded theory study was to examine the experiences of intensive care nurses caring for patients whom they did not believe were going to survive. Participant observation was undertaken in a large intensive care unit (ICU), and formal unstructured interviews conducted with 14 qualified nurses, in order to discover the nurses' perspectives, dilemmas, and role in caring for hopelessly ill patients and their families. The data were analysed by coding and memos, using a constant comparative analysis. The findings emerged into 11 themes which were condensed into three categories: (i) family separation; (ii) trust; and (iii) family reconnection. The core category is 'reconnecting'; the process by which nurses attempt to overcome the dehumanizing aspects of dying in a technological environment. The context, conditions and consequences of this process are discussed and depicted in a conceptual framework. The conclusions drawn include the idea that nurses require instruction regarding managing death in ICUs to enable as peaceful death as possible, not only for the benefit of the patients and their families, but also for the nurses themselves. PMID- 9355424 TI - Effect of endotracheal suctioning on the intracerebral haemodynamics of patients in fulminant hepatic failure. AB - Fulminant hepatic failure is a severe impairment of liver function in someone who has had previously normal liver function. The patients presenting to intensive therapy units have generally had a rapid deterioration which requires prompt intervention and treatment. The first part of this paper gives a detailed review of the aetiology, physiology and management of the disease process. Medical and nursing care of these patients is critical; any adverse intervention may affect outcome. A literature review and small study of the intervention of endotracheal suctioning and its effects on intracranial pressure and cerebral perfusion pressure is described. PMID- 9355425 TI - A concept analysis of the sensoristrain experienced by intensive care patients. AB - Psychological disturbances that patients may experience during admission to intensive care units (ICUs) can have distressing implications for their emotional and physical integrity, progress and subsequent recovery. It is widely believed by practitioners and reflected in professional literature that these disturbances are precipitated by sensory deprivation or overload in the physical environment of intensive care units. In this paper the sources and mechanism of the sensory imbalances experienced by these patients are examined. A new concept- sensoristrain--has been developed in an attempt to promote awareness and improve understanding of the phenomenon among nurses. Once this has been achieved, assessment and identification of patients at risk are optimized and appropriate interventions can be formulated. Using an eclectic approach to analyse sensoristrain, both causes and effects of the phenomenon have been identified from the literature. This information has been combined with practical examples in the development of a model of the concept sensoristrain. The paper concludes by outlining the resulting implications for nursing practice, which may be used to guide future research both in concept development and identification of effective prevention of the phenomenon conceptualized and interventions if it occurs. PMID- 9355426 TI - Ecstasy-induced rhabdomyolysis and its role in the development of acute renal failure. AB - Ecstasy (MDMA) is widely used within the dance scene of the 1990s. In this article the growing trend of ecstasy usage among teenagers is considered, in particular, one of its potentially devastating side-effects, rhabdomyolysis. From consideration of the pathophysiology of rhabdomyolysis and its role in the development of acute renal failure, it becomes apparent that preventative measures and prompt treatment can prevent patients developing acute renal failure as a consequence of rhabdomyolysis. PMID- 9355427 TI - Cardiac electrophysiology studies and ablation procedures: a literature review. AB - This literature review is focused on issues related to the development and usefulness of cardiac electrophysiology studies and ablation procedures. During the past decade, the efficacy of these developments has been proven. The resultant emergence of specialists trained as electrophysiologists represents an important milestone in the advancement of cardiology departments internationally. The majority of research papers on the subject have been published within the past 5 years. Most of the research has evolved from North America and Britain. On searching the literature, it was found that many gaps remain. There is a striking dearth of documented data regarding electrophysiology studies and ablation therapy within the Irish medical and nursing literature, and no previous literature review on the topic was found in a wider search. This paper provides an overview of the strengths and weaknesses associated with these procedures. PMID- 9355428 TI - Aspects of pulmonary artery catheterization in critical care. AB - The concept of floating a balloon-tipped catheter into the pulmonary artery was first described in 1970 by Swan et al. Since then, many issues have surrounded the use of these catheters. Of particular concern for many physicians was the incidence of complications associated with use of the catheters. A group of clinicians have endeavoured to show the usefulness of the pulmonary artery catheter (PAC), but showing significant improvement to patient outcome has proved difficult. Physicians and nurses have demonstrated poor knowledge and skills associated with use of the catheter which must be overcome before conclusive benefits of the catheter can be demonstrated. Training nurses to use a PAC correctly has been highlighted in reducing the number of technical problems associated with the catheter, which in turn improves the accuracy of haemodynamic data obtained. Unfortunately, training programmes are few and far between, and this is an issue that must be addressed by critical care nurse managers. In this review of the literature regarding PACs and their use in the care of the critically ill patients, the role of the nurse is discussed with recommendations for practice. PMID- 9355430 TI - Scholarship in nursing. PMID- 9355429 TI - Paracetamol poisoning. AB - Paracetamol is a common cause of fatal self-poisoning in the UK every year. Despite this, it continues to be sold freely without medical supervision and can be found in quantity in most household medicine cabinets. PMID- 9355431 TI - The role of the nurse in food service: a literature review and recommendations. AB - In the current economic climate increasing numbers of hospitals are outsourcing arrangements for food preparation and service. There is debate among health professionals about the most appropriate role, if any, of nurses in feeding patients. This paper reports on a comprehensive review of the literature over the last 30 years pertaining to patient nutrition and food service in hospital. It is apparent that many patients are admitted in a malnourished state and that deterioration of nutritional status frequently occurs during hospitalization. At the same time there is evidence that appropriate and prompt attention to patient nutrition by nurses and other health professionals has a positive influence on patient outcome and hospital costs. The implications for nursing are discussed. PMID- 9355432 TI - Topical anaesthesia in haemodialysis: evaluation of topical anaesthesia with lidocaine during vascular access in children undergoing long-term haemodialysis for chronic renal failure. AB - Children with chronic renal failure are frequently exposed to painful invasive procedures. Ointments containing lignocaine-prilocaine, when used before intravenous punctures, have been reported to lessen the pain sensation. The aim of this study was to interpret the effectiveness of lidocaine 2.5% ointment in preventing pain when used before venous and arterial punctures in children undergoing a haemodialysis programme for chronic renal failure. Eight children were included in this study. The pain level was identified by the patients using a linear analogue scale. When topical anaesthetic and placebo were compared, there was no statistical difference in interpretation of pain during arterial (P > 0.4), venous (P > 0.375) or both (P > 0.4) procedures. We conclude that lidocaine 2.5% ointment is not effective in preventing pain in children undergoing long-term haemodialysis. In these patients some other factors, like psychological factors, puncture technique and needle size must be taken into consideration for the prevention of pain. PMID- 9355433 TI - Feelings of anger among caregivers of patients with Alzheimer's disease. AB - The purpose of this article is to examine the caregiver's feelings of anger, the causes of these feelings as described by the caregivers and their reactions. Twenty female caregivers of Alzheimer's disease patients participated in a support group at a psychogeriatric centre for 3 years. The group was directed by a registered nurse and a social worker. Frequently, anger was raised as a major issue in the caregiving. The group directors constructed two models that emerged from the support group's meetings. The first model showed the sources of the anger felt by the caregivers and the second model showed coping strategies. The caregivers said that the most infuriating behaviours of patients Alzheimer's disease were: aggressive behaviour, sleeplessness and verbal or behavioural repetition. The caregiver's reactions to the two models are discussed in this paper together with their feelings, which expressed by the women in their own words. PMID- 9355434 TI - An evaluation of the accuracy and efficiency of a school screening model that uses a questionnaire. AB - The role of the school nurse is changing to meet the increasing needs for health promotion and health education. However, the evolution is being hampered by the inefficient work practices involved in undertaking some of the more traditional tasks such as school screening. A survey questionnaire model of screening was developed and trialed with a sample of students and parents and then compared with the results of the more traditional one to one screening programme. The questionnaire survey model was evaluated for accuracy and efficiency against the traditional screening model. The questionnaire method exhibited a relatively low error rate and required one third of the time to complete when compared with the traditional screening programme. The implications of these findings as well as the advantages and limitations of each model are examined in the discussion. PMID- 9355435 TI - Practice patterns of Finnish public health nurses. AB - The aim of this study was to: (i) describe the practice patterns of Finnish public health nurses during the development of primary health care based on the 'population responsibility' principle; and (ii) to identify the relation of public health nurses' task division models and types of community (rural/town) to their practice patterns. A detailed recording of public health nurses' home and clinic visits was developed and used during three annual study periods in 8-9 health centers by 93-118 public health nurses. The total number of recorded visits varied annually from 4842 to 6841. Statistical significance was determined by Chi-squared. Significant differences were found between the three study periods and also in the practice patterns of public health nurses (PHN) with different task division models and of PHN working in different types of community. However, the short study periods limited the practical importance of these findings. PMID- 9355436 TI - Discovering nursing students' understandings about aseptic technique. AB - This phenomenographic study explored understandings about aseptic technique held by second year undergraduate nurses. Data analysis revealed a range of understandings about aseptic technique from simple to complex, referred to here as global, procedural and principled. The implications of these findings are discussed in relation to safe and effective nursing practice. PMID- 9355437 TI - Knowing the patient and the impact on patient participation: a grounded theory study. AB - Currently, there is an emphasis on patient participation. Studies have demonstrated that when patients participate in their care, they experience positive outcomes such as greater satisfaction with care and decreased vulnerability. Nurses are of the opinion that patients should participate in their care. Literature and research indicate, however, that there is an incongruence between nurses' pro-participatory attitudes and actual practice. Grounded theory was used to identify factors relevant to patient participation from the nurses' and patients' perspectives. Using purposive sampling from four hospitals in Western Australia, in-depth interviews and participant observation were conducted. The constant comparative method of analysis was used. As this study is in progress, the core category and sub-categories are still being refined. One category that has emerged strongly is 'knowing the patient' and the outcome on patient participation. This paper will discuss this category from the nurses and patients perspective including recommendations. PMID- 9355438 TI - An exploration of the way in which the concept of patient advocacy is perceived by registered nurses working in an acute care hospital. AB - The purpose of this study was to provide a beginning exploration of the way in which registered general nurses perceived the concept of patient advocacy. A qualitative approach was taken using the techniques of the grounded theory method. A volunteer sample of eight registered nurses working in an adult acute care ward of a major metropolitan hospital were interviewed using an semi structured format. Areas explored during the interviews included personal definitions of patient advocacy, elements of the advocate role, and the rationale for the nurse acting as patient advocate. Data analysis commenced during the data collection phase, where initial interviews were transcribed, coded and beginning categories were identified. The findings indicate that for the participants, advocacy is based on respect for the person, and acknowledgement of human rights. The quality of the relationship between the nurse and the patient appears to be the basis for the advocate role. Three initial categories emerged that describe the major elements of the participant's conceptualization of patient advocacy. These are, informing, supporting and representing. PMID- 9355439 TI - Changing handover practices: one private hospital's experiences. AB - The handover practice has long been an important component of clinical nursing practice allowing nurses to exchange relevant client information from one shift to the next and ensure continuity of patient care. Traditional approaches have seen nursing handovers taking place in a room away from general ward activity. Oncoming nursing staff receive the information verbally from nurses on the previous shift about all patients within the ward or unit. This practice has been proven over time to present difficulties and consequently, many hospitals are choosing to adopt models that better address current needs. This analysis describes the creative approaches taken by one private hospital in modifying handover practices with the view to reduce time and increase overall efficiency and effectiveness, whilst ensuring that staff and ward requirements are considered. The study highlights how action research principles can be applied to introduce change into the clinical practice environment. PMID- 9355440 TI - Innovations in community health nursing: examples from practice. AB - The Ottawa Charter for Health Promotion is cited internationally as an appropriate conceptual framework for healthcare service delivery yet the literature reveals minimal evidence of nursing services interpreting and applying the Ottawa Charter strategies into nursing practice. Nurses with the community services of Noarlunga Health Services and the Drug and Alcohol Services Council of South Australia, however, do use the strategies to plan and implement their services. The Ottawa Charter strategies of developing personal skills; creating supportive environments; strengthening community action; building healthy public policy; and re-orienting services in the interest of health can be used as a tool to assist nurses to identify the purpose of their interventions and select a comprehensive range of nursing actions which address the needs of individuals while acknowledging the broader determinants of health. This article presents a nursing analysis of the Charter and provides examples of how the strategies are used to influence nursing practice in both organizations. The examples provided from the two different nursing services also demonstrate the adaptability and relevance of the strategies to diverse community nursing practice settings. PMID- 9355441 TI - Helping older people and their families to cope with grief. PMID- 9355442 TI - Daily administration of granulocyte colony stimulating factor using a subcutaneous butterfly. PMID- 9355443 TI - [Health and nursing care meeting 1997. Both serious and funny]. PMID- 9355444 TI - [Health and nursing care meeting 1997]. PMID- 9355445 TI - [Working environment for pregnant women]. PMID- 9355446 TI - [Acting instead of talking gives a different perspective]. PMID- 9355447 TI - [Knowledge in multiple extremity abnormalities. Even today children are born without arms and legs]. PMID- 9355448 TI - [Desire for love. Problems in sex- and lovelife]. PMID- 9355449 TI - [Desire for love. We need to have desire]. PMID- 9355450 TI - [Desire for love. Give yourself and your partner a week of love! The art of being egoistic in life together. Interview by Ingela Radestad]. PMID- 9355451 TI - [Desire for love. Midwife and sexology]. PMID- 9355452 TI - [Desire for love. Geni(t)al capital]. PMID- 9355453 TI - [Desire for love. Men don't want to pull their pants down]. PMID- 9355454 TI - [Desire for love. Report from the condom horizon]. PMID- 9355455 TI - [World congress in obstetrics and gynecology]. PMID- 9355456 TI - [Insufficient control of a new spiral]. PMID- 9355457 TI - [Problems with spirals]. PMID- 9355458 TI - [Delivery in the year 2000--team work obstetrican-midwife]. PMID- 9355459 TI - [Midwives' need--of men! Observations from two "converts"]. PMID- 9355460 TI - Child and adolescent mental health: a global perspective. PMID- 9355461 TI - Use of electroconvulsive therapy with children: an overview and case report. AB - TOPIC: Electroconvulsive therapy (ECT) has become more common in the treatment of adults with refractory mood disorders and psychotic disorders but it remains one of the least common therapies for mental illness in children. In a small number of child psychiatry cases, symptoms are severe and unresponsive to standard pharmacological and other therapies. With these patients, ECT might be helpful. PURPOSE: This article provides an overview of ECT, indications for its use and a case report that illustrates the successful use of ECT with an 8-year-old girl with psychotic depression. Implications for multidisciplinary care are discussed, including preparation of the patient and family, assessment of response to ECT, management of adverse effects, preparation for discharge and discharge care. SOURCES: Existing literature on the use of ECT in adults, adolescents, and children and the clinical experience of providing care to an 8-year-old patient on an acute care inpatient unit. CONCLUSIONS: Nurses and other healthcare personnel should consider ECT in refractory cases of major depressive disorder, bipolar affective disorder, schizophrenia, and other psychotic disorders. PMID- 9355463 TI - Factors in childhood drug and alcohol use: a review of the literature. AB - TOPIC: Drug and alcohol use in children. PURPOSE: To identify the prevalence of drug and alcohol use in children 12 and younger and the factors associated with this use. SOURCE: A review of the literature. CONCLUSIONS: The identified factors include: a) differences in use between boys and girls; b) influence of family members' drug and alcohol use; c) influence of peers; d) the child's self-esteem; and e) the child's knowledge about drug and alcohol use. Awareness by child psychiatric nurses of the identified factors is crucial to prevent, assess, and treat the problem of drug and alcohol use by children. PMID- 9355462 TI - Teaching kids to cope: a preventive mental health nursing strategy for adolescents. AB - TOPIC: The theoretical base, implementation, and effectiveness of Teaching Kids to Cope (TKC). PURPOSE: To inform clinicians about the benefits and uses of TKC, a 10-week psychoeducational group intervention designed to enhance the coping repertoire of adolescents. SOURCE: Authors' clinical experience. CONCLUSIONS: Following the TKC intervention, students demonstrated less depressive symptomatology and improved coping behaviors. Further research is needed to test the intervention on a larger group and to determine its effectiveness. PMID- 9355464 TI - Self-mutilating behavior. AB - TOPIC: The phenomenon of self-mutilating behavior. PURPOSE: Self-multilating behavior (SMB) creates serious management problems in psychiatric settings and typically begins in adolescence. In order to plan and provide effective nursing interventions, it is necessary nurses be knowledgeable about the phenomenon. This paper provides definitions, historical views, and types of SMB, along with a description of current research and speculations about intent. This framework offers a background to nursing interventions focused on identifying at-risk adolescents and treating this bewildering behavior. SOURCES: Published literature. CONCLUSIONS: Mental health professionals find treatment of SMB a challenge and nursing is in a unique position to provide effective interventions. PMID- 9355466 TI - Summer conference season. PMID- 9355467 TI - The therapeutic use of music in a care of the elderly setting: a literature review. AB - This paper reviews recent literature concerning the use of music and music therapy in health care. Focusing particularly on the elderly, the use of music in relation to patients with dementia and Parkinsonism is examined. Brief reference is also made to the use of music in pain control. Although in this case, literature is not specific to care of the elderly settings, the results are still relevant to gerontological nursing. Projects which achieved positive results in controlling pain perception could be transferable to a care of the elderly scenario, where chronic pain is often part of daily life. PMID- 9355468 TI - Defining 'competency' in nursing (Part I): A policy review. AB - How the nursing profession in England defines the basic competency of the registered general nurse is crucial for the safety of the patient and the protection of the nurse. It is also essential for determining advanced practice, extended practice and specialist practice in nursing. An historical overview of documentation from the United Kingdom Central Council for Midwifery and Health Visiting, the English National Board for Nursing, Midwifery and Health Visiting and the Royal College of Nursing shows a profound change in educational policy during the 1980s regarding the interpretation of nursing competency. The effects of this policy change on the current methods developed to define, teach and test nursing competency are examined. Preliminary conclusions show shortcomings and uncertainty in the present preparation and evaluation of nursing competency. PMID- 9355469 TI - Changes in practice nursing: professionalism, segmentation and sponsorship. AB - In the wake of recent government policies and legislation, practice nursing has undergone major changes. Not least of these has been the rapid growth in the number of practice nurses. In addition, their work role has grown greatly over recent years. This has given rise to the question of whether their professional development now justifies their status as autonomous nurse practitioners. This and related issues are explored in the empirical part of this paper, which focuses on the changes in the work and responsibilities of practice nurses in two industrial towns in the English Midlands. PMID- 9355465 TI - The use of traditional neuroleptics in children and adolescents. AB - Neuroleptics are used for a wide range of neuropsychiatric conditions in children and adolescents. Although the body of evidence from controlled studies is limited, there is some information upon which to base clinical practice (McClellan & Werry, 1994). These medications can be useful in psychosis, tic disorders, severe hyperactivity, agitation, and aggression. Nonetheless, the neuroleptics can have both short-term and long-term adverse effects. Contemporary practice in child and adolescent psychiatry requires knowledge about the rational uses, dosing, and side effects of these medications. Prior to initiating a trial with a neuroleptic medication, the risks, benefits, and alternatives should be reviewed with the child and family. PMID- 9355470 TI - 'A thorny problem for feminism': an analysis of the subjective work experiences of enrolled nurses. AB - Feminist methodology has been used in this study to investigate the subjective work experiences of 19 enrolled nurses. Reflexive conversational techniques enabled participants to relate autobiographical narratives examining their experiences of working in a nursing hierarchy. Two themes are reported: 'nursing identity' and 'exploitation'. Through an analysis of these themes insight has been gained into the manner in which nurses inter-relate. The study demonstrates how the correlation between rank, status and expertise within the nursing hierarchy may have negative consequences for patient care. PMID- 9355471 TI - An exploratory study from the patients' perspective of living with allergies. AB - Allergic disease is one of the most prevalent chronic medical conditions in the world. Allergen avoidance has been accepted as a form of treatment for allergic disease; however, the success of treatment is often dependent on how patients choose to manage their condition. The purpose of this study was to explore how allergic conditions affect the lives of allergy sufferers and what information they believe would be useful to other allergy sufferers in the management of their allergic condition. Patients suffering from non-life-threatening allergies stated that their allergy affected many aspects of their life, such as their work, their social life, their emotional state, their physical appearance and, hence, their interactions with others. The chronic symptoms of their condition caused the greatest concern to patients. Many of these participants 'accepted their condition' and undertook controlling-measures as part of their daily life. In contrast, patients suffering from life-threatening allergies stated that their allergy did not affect their life. Knowledge of their allergic condition was more important than acceptance. The findings elicited from allergy sufferers identified how nurses can educate patients about their condition and assist them in learning to live with their condition. PMID- 9355472 TI - A study of nurses' views about the prevention of nosocomial urinary tract infections. AB - This study sought to discover the contribution of nursing practice to the prevention of hospital-acquired or nosocomial urinary tract infections (NUTIs), the most commonly occurring nosocomial infection. Seventy-five per cent of such infections are associated with urethral catheters. The practices of nurses who are caring for patients on a 24 h basis would appear to be fundamental to achieving any reduction in the incidence of NUTIs. This qualitative study utilized unstructured interviews to explore the views of 12 registered nurses about three key issues: first, what care do nurses give with the aim of preventing catheter-associated NUTIs; secondly, what improvements in practice would further prevent catheter-associated NUTIs; thirdly, what do nurses see as constraints to the prevention of catheter-associated NUTIs? The nurses identified many of the measures that were cited in the literature as effective for preventing NUTIs; however in reality, they stated that their practice differed because of a lack of time to give care and to update themselves. The consequences of under-staffing were that junior and temporary staff (whose competence in preventing NUTIs was questioned) worked unsupervised. Those interviewed identified feelings of powerlessness in effecting preventative measures, and identified not only the role of medical staff in influencing NUTIs but also their inconsistent approach to care. All these forces effectively limited the nurses' ability to prevent NUTIs. The study is concluded with recommendations for changes in practice and further research. PMID- 9355473 TI - Implementation of clinical supervision in a medical department: nurses' views of the effects. AB - The purpose of this pilot study was to investigate nurses' views of the effects of clinical supervision in terms of its influence over their working situation, as well as their satisfaction with their working milieu. The nurses who took part in an education programme and a clinical supervision programme worked on two wards in a medical clinic. Nurses answered a questionnaire measuring the psychosocial environment. Data were analysed by means of descriptive statistics. Results showed that the nurses felt more confirmed in their work and more satisfied with the information given after 9 months of clinical supervision. Results point to the need for further investigations concerning clinical supervision as a method of achieving job satisfaction among nurses. PMID- 9355474 TI - Evaluating the introduction of primary nursing: the use of a care plan audit. AB - A care plan audit was carried out as part of an action research project involving the introduction of primary nursing. The audit tool was based on the Roper, Logan and Tierney Activities of Living model and the nursing process. The audit showed that few changes in documentation had taken place as a result of the introduction of primary nursing. The volume of communications had increased but much of this was not documented on care plans. Other positive changes as a result of introducing primary nursing were found, and both patients and nurses were aware of these. PMID- 9355475 TI - Survivorship and breast cancer: the psychosocial issues. AB - This article addresses some of the psychosocial issues concerning survivors of breast cancer. Issues relating to employment and social support are discussed. Social support is also related to the significant other. Recommendations for nursing practice and research are made, including the need for a unilateral definition of 'survivor' and for attitudinal changes. As the number of survivors increase, psychosocial barriers need to be diminished by nursing practice and research. PMID- 9355476 TI - Better late than never? An evaluation of community nursing services for children in the UK. AB - The development of community nursing services for children in the UK was first suggested by Platt in 1959; until recently, however, such services have been rare and children have spent many unnecessary and potentially traumatic days in hospital. This paper considers the development of community nursing services for children in the UK. It identifies factors which have promoted the growth of these schemes, and examines the effectiveness of services from professional, managerial and economic perspectives. The viewpoint of the consumer--the child and family themselves--is also considered. PMID- 9355478 TI - A cultural look at aging. PMID- 9355477 TI - Stress and fitness in ward-based mental health nurses. PMID- 9355479 TI - The family's role in long-term care. AB - Long-term care provided by family members is the central care of our current health care system. The purpose of this article is to review the family's role in long-term care. Issues such as cost containment and inequities in our current social policy are reviewed. Suggestions for future directions in social policy are presented. PMID- 9355480 TI - To grandmother's house we go ... and stay. Children raised in intergenerational families. AB - An increased incidence in child abuse and neglect has resulted in a dramatic rise in the number of grandparents raising grandchildren and great-grandchildren. Grandchildren raised by grandparents often suffer from emotional and behavioral problems due to prior abuse, neglect, and abandonment. Grandparent caregivers experience increased health problems, psychological distress, and social isolation related to their roles as primary caregivers of children. Grandparents who become caregivers of grandchildren face increased financial responsibilities at a time in their lives, close to or at retirement, when income is dramatically decreased. PMID- 9355481 TI - Learning from other lands. Caring for elderly demented Koreans. AB - The aims of the study reported here were to describe the socio-demographic characteristics of caregivers of demented elders in Korea and their care recipients and to compare the positive and negative meanings and outcomes of the caregiving experiences of caregivers who had admitted their elderly demented relative to a nursing home (G1: n = 24) and caregivers still caring for their elderly demented relatives at home (G2: n = 30). Most caregivers were female (80%), married (89%), and related to the care receiver as daughter-in-law (39%), daughter (22%), wife (15%), son (13%), or neighbor (6%). Social class differences were found between the home care and nursing home groups: the upper classes were significantly more likely to have placed their demented elder in a nursing home, whereas the low social classes were more likely to keep taking care of their demented elder at home instead of placing them in a nursing home. Caregivers who had admitted their relative to a nursing home (G1) reported significantly more difficulties from disturbed sleep, disrupted children's studies, and limited personal life when they were caring for the elder at home (p < .05). Caregivers in the home care group (G2) had significantly greater satisfaction in serving as a model for their children and practicing religion (p < .05), and they also reported a better relationship with the care receiver than those who have placed their demented elder in a nursing home, although the difference in this case was not significant. PMID- 9355482 TI - Trends in aging care in Scotland and Scandinavia. AB - The proportion of older adults in Western European countries, as in the United States, continues to increase rapidly. Faced with geriatric care dilemmas decades earlier, however, these countries have had more experience on which to base the development of community-based, integrated care systems for the elderly. This article provides observations from a 1993 World Health Organization Fellowship study of long-term care facilities in four European countries: Scotland, Sweden, Norway and Denmark. Several emerging trends in geriatric care documented in the literature were confirmed. These included: moratoria on institutional long-term care, emphasis on informal care and support, provision of 24-hour assistance in the home, care management to individualize care, and an expanded set of providers within integrated delivery systems. PMID- 9355483 TI - Nursing and health care for an aging society: the case of The Netherlands. AB - 1. Demographic developments in European and western countries are not unique. Nurses should embrace the opportunity to learn from each others' care and research initiatives for the elderly population. 2. Current transitions in health care systems should be seized as an opportunity to further establish and develop the nursing profession, for example, through joining and initiating multidisciplinary initiatives. 3. The aging population is the fastest growing population in a number of countries. The training, recruitment, and retainment of nursing staff are key to continuously provide high quality care for the elderly. PMID- 9355484 TI - Complementary and alternative therapies for our aging society. AB - Gerontological nurses are dynamic individuals who recognize the necessary changes needed to reform a biotechnology-driven health care system to become a relationship-centered, care-driven healing system with elders. They must continue to take action to enhance caring and healing at personal levels and in various professional levels of clinical practice and health care reform. As gerontological nurses pursue personal, clinical, educational, and research approaches to holistic nursing and caring-healing modalities, they increase their knowledge and skills of holism and the complex body-mind-spirit interconnections, and provide relationship-centered care (Tresolini, 1994). Gerontological nurses then truly blend the art and science of caring with elders, and lead to a deeper understanding of healing as a lifelong journey into wholeness (Achterberg, Dossey & Kolkmeier, 1994). PMID- 9355485 TI - What's hot, what's not, in dementia drugs? Part I: Cholinergic therapies. PMID- 9355486 TI - Building community: developing skills for interprofessional health professions education and relationship-centered care. PMID- 9355487 TI - Differential diagnosis of decreased vision: a case study. PMID- 9355488 TI - Is it Reiter's syndrome? PMID- 9355489 TI - Depression. U.S. Public Health Service. PMID- 9355490 TI - Urticaria: a nurse practitioner's approach. PMID- 9355491 TI - 1995-96 Kansas Nursing Wage Survey. PMID- 9355492 TI - Kansas nursing occupational projection 1995-2005. AB - The Kansas State Department of Human Resources recently released a study of occupational employment throughout the state. The data was compiled using staffing pattern employment from the 1992, 1993 and 1994 occupational employment statistics program survey of industries, along with base year 1993 average annual employment for Kansas industries. Projected demand for occupational employment is significantly affected by the projected employment for industries. PMID- 9355494 TI - Protect those ears before it is your loss. AB - This article describes the different forms of hearing loss. Causes of sensiorneural and conductive hearing loss are identified. Hearing loss is rapidly affecting the population. Noise not only affects the working population but can adversely affect persons who are surrounded by excessive noise in the environment from recreational activities. Hearing loss affects one in ten persons and is considered one of the top ten occupational hazards. PMID- 9355493 TI - Today's nurse plans for tomorrow. PMID- 9355495 TI - Family legacy. PMID- 9355496 TI - Myths & facts ... about fire safety. PMID- 9355497 TI - Stethoscope safety tips. PMID- 9355498 TI - Maintaining a closed urinary drainage system. PMID- 9355500 TI - Colorado nurses' trial. Learning from medication errors. PMID- 9355499 TI - Managing pain in a drug-abusing patient. PMID- 9355501 TI - Helping patients find health information on the Web. PMID- 9355502 TI - Stockpiling painkillers. PMID- 9355503 TI - Actionstat. Hydrofluoric acid burn. PMID- 9355504 TI - Mitral valve prolapse revisited. PMID- 9355505 TI - Carol's dove. PMID- 9355506 TI - Saving lives with automated external defibrillators. PMID- 9355507 TI - Caring for Jason ... one day at a time. PMID- 9355508 TI - Going home. Is home health care for you? PMID- 9355509 TI - Caring for patients with pancreatitis. PMID- 9355510 TI - Teaching your patient to use a metered-dose inhaler: the direct route for asthma therapy. PMID- 9355511 TI - Focusing on the dangers of D5W. PMID- 9355512 TI - Job safety hand book. PMID- 9355513 TI - 8 signs of poor communication. PMID- 9355514 TI - Lung volume reduction surgery: making room for easier breathing. PMID- 9355515 TI - Venipuncture tips for geriatric patients. PMID- 9355516 TI - Teaching your patient self-documentation skills. PMID- 9355517 TI - Nicotine polacrilex gum. PMID- 9355518 TI - Returning the love. PMID- 9355519 TI - Heating devices. How to avoid burns. PMID- 9355520 TI - Making your voice heard on legislative issues. PMID- 9355521 TI - Watching out for Edna. PMID- 9355523 TI - Human rights and Europe confused. PMID- 9355522 TI - MRSA. AB - All nurses are likely to encounter methicillin-resistant Staphylococcus aureus (MRSA) at some time in their career, and they are also likely to observe that approaches to containing it vary from hospital to hospital and within different care settings. This Learning Unit aims to explore these apparent contradictions, and identify the elements of practice common to all care settings. The associated television component makes clear that the needs of the patient are central to all approaches to tackling MRSA. The themes of this workbook are based on the settings in which care is given. The settings are important as predictors of the acquisition and spread of MRSA, because they indicate the vulnerability of the patients within them, and the kind of treatment and care the patients are likely to receive. Susceptibility to MRSA is linked to surgery, to the use of invasive devices and to the immune state of the patient. The physical environment, the fixtures and fittings, furniture and equipment, are seldom implicated in the spread of MRSA. MRSA lives on skin, and it is skin that helps it to spread, particularly on the hands of healthcare workers. The focus of this workbook is on the simple, everyday practices which interrupt this method of spread. This Unit is relevant to the UKCC Professional Development Categories: Care enhancement and Reducing risk. PMID- 9355524 TI - Legal penalties and HIV. PMID- 9355525 TI - Quality costs. PMID- 9355526 TI - Mind your language. PMID- 9355527 TI - Perspectives-making sense. PMID- 9355528 TI - Changes or cuts? PMID- 9355529 TI - Romania revisited. PMID- 9355530 TI - International strength. Interview by Norah Casey. PMID- 9355531 TI - Assessment of need for health and social care. AB - This report highlights two studies into how clients' health and social care needs are assessed (Camey et al 1996). The studies were carried out in Scotland, by researchers at Glasgow Caledonian University, and in Northern Ireland, in a joint project between Queen's University, Belfast, and the University of Ulster. PMID- 9355532 TI - Diversional group therapy: a study of effectiveness. AB - This research uses an immediate response questionnaire (IRQ) to measure the general diversional effectiveness of six new recreational and diversional groups in a 38-bedded acute psychiatric unit. The groups were first implemented in September 1996. The results support the implementation and continued use of the new groups and the methods used in facilitating them. PMID- 9355533 TI - Introducing a bereavement support programme in ICU. AB - This paper outlines the development and introduction of a support programme for bereaved relatives in the intensive care unit. The programme is distinguished by nursing follow up of relatives following the patient's death. PMID- 9355534 TI - Telephone assessment and advice: a training programme. AB - Nurses have become increasingly involved in providing telephone assessment and advice, and telephone triage is emerging as an important part of the everyday role that nurses can play in A&E, general practice and other community settings. This article describes a project to prepare staff to fulfil this role. PMID- 9355535 TI - The effects of massage: an holistic approach to care. AB - In this article the authors review the benefits of massage and its relationship and relevance to more orthodox therapies. Most studies of the effects of massage on patients' wellbeing have been undertaken by non-nursing researchers and the authors suggest that more nurse-based research would make an important contribution to an holistic approach to care. Interest in, and the use of, complementary therapies has been growing over recent years. Massage seems to be of particular interest to nurses, involving as it does the close, intimate contact which nurses are often engaged in as part of their day-to-day work with patients. The benefits, and some of the problems, associated with massage are explored below. PMID- 9355536 TI - Practical stoma care. PMID- 9355537 TI - Efficacy of costly AIDS treatment questioned. PMID- 9355538 TI - Nurse on HIV ward found to have TB. PMID- 9355539 TI - Long hours and dirty linen. PMID- 9355540 TI - Reluctant diagnosis. PMID- 9355541 TI - Can't pay, won't pay. PMID- 9355542 TI - False economy. PMID- 9355543 TI - Click to connect. PMID- 9355544 TI - The outer edge of consciousness. PMID- 9355545 TI - A stranger in the family. PMID- 9355547 TI - Where did Mother go? PMID- 9355546 TI - Know how. Vitamins and minerals. Part Six. PMID- 9355549 TI - Skills from a suitcase. PMID- 9355548 TI - Individual responsibility or corporate culture? PMID- 9355550 TI - Knowing me, knowing you. Interview by Eileen Fursland. PMID- 9355551 TI - It's clappy but are we happy? PMID- 9355552 TI - Compression therapy for venous ulcers: a systematic review. AB - This article summarises the findings of a systematic review carried out by the NHS Centre for Reviews and Dissemination and the Center for Evidence-Based Nursing. Based on a rigorous analysis and synthesis of available research, the review concludes that training in the assessment of leg ulcers and the correct application of compression therapy appears to be more important than the particular products used. Recommendations for audit are also reported. PMID- 9355553 TI - Factors influencing the experience of pain. AB - The experience of pain is influenced by a range of complex factors, all of which need to be taken into account if appropriate and effective care is to be provided. In this article, the third in a series of six, a range of factors that need to be considered in assessing a person's pain is discussed. These include the effect of age, previous experience of pain, gender and culture. The fourth article in the series will be published in next week's Nursing Times. PMID- 9355554 TI - Cardiovascular assessment--Part 2. AB - This is the fifth part of the Care is Critical series, which looks at the more complex assessments and technological interventions nurses may now have to deal with on general wards or in the community. This article aims to provide a more in depth understanding of assessing patients' cardiovascular function in relation to low cardiac output states in two types of patients--the post-operative patient with hypovolaemia and the patient with chronic heart failure. PMID- 9355556 TI - Stoma care: teaching patients to cope. PMID- 9355555 TI - A survey into the smoking habits of nursing students. PMID- 9355557 TI - The challenges of health care restructuring. AB - Since the late 1980s, virtually every developed, and many developing, countries have re-examined the structure of their health care systems. Health care reform has become a truly global phenomenon with considerable potential for cross-nation lesson-learning. In countries where the state has been the central actor in the health sector, its role is being reassessed and, in some cases, reconfigured. The introduction of market principles to health care is a feature of many countries: market romantics believe markets in health care will improve efficiency, empower consumers, control costs, and overthrow monolithic bureaucracies. But will they? The evidence, such as it is, suggests otherwise. The greatest pressure for change and for introducing markets into health care has been in the relative role of the private sector in the operation, and in some countries also the financing of health care services. But it is not a simple case of the state versus the market. The issues are much more complex and various hybrid models are emerging involving some sort of public-private mix. The move is towards greater diversity and pluralism, an inevitable consequence of which is growing fragmentation in the funding and provision of care with all the associated on-costs in terms of increased coordination and management that this entails. The policy aim is to harness the benefits of market behaviour without also adopting the inherent weaknesses of markets with regard to questions of distributive justice and equity. PMID- 9355559 TI - The driving force. PMID- 9355558 TI - A few home truths. PMID- 9355562 TI - "ER" TV show demeans nursing. PMID- 9355561 TI - ONA pushes powdered latex ban. PMID- 9355560 TI - A place in the record books. PMID- 9355563 TI - Proposed legislation attacks labor unions. PMID- 9355564 TI - Hospice nurses unite, win on patient risks. PMID- 9355565 TI - Latex: a deadly allergy. PMID- 9355566 TI - Patient protection update. PMID- 9355567 TI - Genetic education and counseling. AB - This special issue of Patient Education and Counseling on genetic education and counseling provides an overview of studies and findings in this field. It features a mixture of papers dealing with five different topics related to several psychosocial aspects of genetic education and counseling. Attention is paid to new issues in counseling for hereditary cancer and Huntington Disease. Articles are presented on information recall of counseled individuals, the use and impact of genetic services on counselees (acceptance of testing; knowledge of inherited cancer susceptibility; risks of genetically testing children). Also topics are addressed with respect to the counselor (neutral attitude; understandable language; information recall; satisfaction with the services provided by the genetic counselor). Furthermore, recommendations are discussed for screening practices for women with a family history of breast cancer, and in addition, the effectiveness of genetic counseling is addressed. In conclusion several suggestions for future research are given. PMID- 9355568 TI - Facts in cancer genetics. AB - During the past decade molecular biology and molecular genetics have greatly increased our understanding of the basic mechanisms in cancer development. The essential outcome of these molecular studies is that cancer can be considered as a genetic disease of cells. Both the non-hereditary (sporadic) cancers, as well as the hereditary forms of cancer are caused by genetic accidents that perturb the complex and delicate cellular growth control systems. Thus, at the molecular level, no principal difference exists between hereditary- and non-hereditary forms of cancer and it can be stated that both at the level of the single cell as well at the level of the individual, cancer is a genetic disease. Whereas in hereditary cancer, the risk gene is passed through the germline to the next generation, in sporadic cancer, a cancer cells passes its abnormal genes to its daughter cells at cell division. It is therefore not surprising that one of the main priorities in cancer research today is the identification of the culprit genes and characterizing the function of their normal products. Genes associated with hereditary cancer syndromes essentially encompass two classes of genes viz. tumor suppressor genes and genes controlling genomic stability (DNA-mismatch repair genes). Although germline mutations in these susceptibility genes are associated with significantly increased cancer risk, even up to 90%, additional genetic factors and interaction with environmental factors eventually determine if a carrier of a germline mutation will develop cancer. PMID- 9355569 TI - Psychological issues in cancer genetics: current research and future priorities. AB - There has been a rapid expansion of genetics research in the field of cancer since cancer predisposing genes are now known to cause a proportion of common cancers as well as rarer cancer syndromes. As a result, the psychosocial impact of being at high risk of cancer has become a focus of evaluation, and studies are being reported which set out to evaluate both the uptake and psychological outcome of genetic counselling, testing and surveillance. Available data concerning psychological aspects are reviewed, including for example, possible implications of genetic testing, attitudes and uptake of breast screening and accuracy of women's risk estimates. Work is in progress to assess the more controversial areas of prophylactic mastectomy, and chemoprevention. Other research examines the longer term impact of belonging to a Cancer Family, and of interventions offered to high risk families. This is crucial since the uptake of counselling and testing is likely to be much greater in cancer prone families than those with other genetic disorders, yet detection and prevention strategies are still unevaluated for important genetically determined cancers such as breast cancer. PMID- 9355570 TI - Psychological aspects of genetic counselling: a review of the experience with Huntington's disease. AB - Presymptomatic DNA-testing for adult-onset diseases has serious psychological consequences. Here the psychological consequences of presymptomatic DNA-testing for Huntington's disease are reviewed. Both carriers and non-carriers experience emotional reactions after disclosure of their test result. However, up to today no long-term adverse emotional consequences have been revealed. Future research on other adult-onset genetic diseases should provide information about the reactions of children. In genetic counselling, attention should be paid to the reactions of people with a decreased risk. Genetic counselling must focus on the whole family and not on the individual applicant. PMID- 9355571 TI - Acceptance of genetic testing in a general population: age, education and gender differences. AB - The aim of the study was to analyze effects of age, education and gender on acceptance of genetic testing. Subjects, n = 1967 aged 15-69, were a stratified random sample of the Finnish population. One thousand, one hundred and sixty nine subjects, 530 men and 639 women, returned the questionnaire. The majority of the respondents approved of the availability of genetic testing. Young, aged 15-24, were more favourable towards testing and more willing to undergo suggested tests, but they were also more worried than others about the misuse of test results. Men aged 45-69 with only basic education were more in favour of mandatory genetic testing than other respondents. Respondents with university education were more critical towards genetic testing and expressed their worry about eugenics more often than other education groups. In conclusion, there are age, education and gender related differences in acceptance of genetic testing which need to be taken into account when considering screening programmes and informing the public. PMID- 9355573 TI - Psychological risks of genetically testing children for a hereditary cancer syndrome. AB - Parents in families with a hereditary cancer syndrome are often familiar with periodical clinical testing of both themselves and their children. Genetic testing is an additional early diagnostic option that is becoming available for an increasing number of hereditary cancer syndromes. Participants in genetic counseling programs for cancer syndromes are often parents who apply for their children. If a child is identified as a carrier of a specific disease-causing gene mutation, sometimes its parents must decide on when it will be treated can treatment be postponed until expression of the disease or should the child receive presymptomatic surgery? We discuss some of the possible risks of genetically testing children: distress as a result of ambivalent feelings towards testing, preoccupation with disease-related signs, changes in family interactions, the burdening prospect of a future disease and medicalization of the carrier-child. PMID- 9355572 TI - Ethnic differences in knowledge and attitudes about BRCA1 testing in women at increased risk. AB - Informed consent for BRCA1 mutation testing will require adequate knowledge of patterns of inheritance of cancer and the benefits, limitations, and risks of DNA testing. This study examined knowledge about the inheritance of breast cancer and attitudes about genetic testing for breast-ovarian cancer susceptibility in women at increased risk. Knowledge and attitudes were measured in 407 African American and Caucasian women aged 18-75 who had at least one first-degree relative (FDR) with breast and/or ovarian cancer. The average knowledge score was 6.0 out of a total of 11 (S.D. = 2.15). Compared to Caucasian women, African American women had lower levels of knowledge and had more positive attitudes about the benefits of genetic testing. There were no significant ethnic differences in attitudes about the limitations and risks of testing, however, income was negatively associated with this outcome. Ethnic differences in knowledge and attitudes about genetic testing for breast-ovarian cancer risk may be attributable to differences in exposure to genetic information and referral by health care providers. PMID- 9355574 TI - The standard of neutrality during genetic counselling: an empirical investigation. AB - One of the standards for the genetic counsellor's profession is neutrality, which enables clients to decide 'in freedom' what is best for them. However, in a world with divergent and changing values, neutrality may be difficult to achieve. In order to highlight possible biases in counsellors' communications, verbal exchanges during 30 counselling sessions in a clinical genetics centre in the Netherlands were analyzed. The results show that the main background against which the attitudes of both clients and counsellors must be considered is their notion of the rapid development of medical science. Although most counsellors' attitudes clearly reflected a striving for neutrality, some did not always succeed in that: (a) they exceeded the original brief, (b) they implicitly expressed their own opinions and values, (c) they ignored client' objections and (d) they issued directives. Practical implications are discussed. PMID- 9355575 TI - Clinical terminology: anxiety and confusion amongst families undergoing genetic counseling. AB - Genetic counseling is a rapidly expanding, but highly demanding, domain of doctor patient communication. This paper reports results from an ethnographic study of families (n = 30) attending a genetic counseling clinic in Northern England. We suggest that the language used in this particular specialty is often confusing and misunderstood by the families involved. We found that unfamiliar terms may also conjure up also conjure up alarming images. It is important therefore, that physicians, and in particular geneticists, try to use simple, understandable language, and give clear explanations for unfamiliar terms that cannot be avoided. The careful choice of words, and detailed explanation, not only reduces the risks of "labelling" and stigmatization, but may also prevent the unnecessary anxiety experienced by patients when they hear unfamiliar medical terms, such as the eponyms frequently employed by geneticists when giving a diagnosis. PMID- 9355576 TI - Information recall in genetic counselling: a pilot study of its assessment. AB - One of the main aims of genetic counselling is to provide information to patients that they can understand and recall. Measuring information recall is problematic and most studies do not make the comparison between recalled information and the information that is given in consultations. The aim of this study was to determine the validity of using genetic counsellors' reports of information given in genetic consultations as the basis for a measure of patient recall. Counsellors in 35 routine genetic counselling consultations were asked to indicate important information given during consultations by highlighting items in their summary letters sent to patients. This was checked by a researcher against tape-recordings of the consultations. One month later, patients were telephoned by the researcher and asked to recall as much information as they could from the consultation and to rate its importance. Over 95% of the information that counsellors identified as important had been given during the consultation. Patients rated this information as important. Although there was an association between the overall number of items that counsellors and patients judged to be important, patients more frequently judged information about family implications to be important than did counsellors. On the other hand, counsellors more frequently judged information about test, diagnosis and prognosis to be important than did patients. These results suggest that counsellors' summary letters are a valid baseline against which to measure patient recall. This measure will not, however, capture all the information that patients consider important. PMID- 9355577 TI - Genetic counselling: information given, recall and satisfaction. AB - The aims of this study are to categorise the key points given in genetic counselling, assess the amount and type of information recalled, and examine the relationships between counsellees' knowledge, satisfaction with information received, the meeting of expectations, concern and anxiety. Because of the variety of consultations, a knowledge questionnaire of the key points was constructed for each individual counsellee, with acceptable inter-rater reliability. The information items judged to be the key points in the consultations were assigned to 13 different content categories. Thirty-two counsellees were sent a questionnaire assessing knowledge and other outcomes two to four weeks after attending a genetic counselling consultation. The mean percentage of key points recalled correctly was 76% (s.d. 17%) with 100% recall for family issues and 68-78% recall for genetic or medical information. Knowledge was not associated with satisfaction with information received nor with level of concern or anxiety following genetic counselling. These results suggest the importance of assessing multiple outcomes of genetic counselling. PMID- 9355578 TI - Genetic counseling for hereditary cancer: a pilot study on experiences of patients and family members. AB - Recent advances in the identification of genetic abnormalities associated with certain types of cancer have stimulated the development of screening and counseling programs for hereditary and familial forms of cancer. In 1995, such a program was established in a collaboration between three familial cancer clinics in Amsterdam. Given the potential impact of genetic screening and counseling on the psychosocial health of participants, it was considered essential that the program be evaluated from its inception to determine the participants' satisfaction with the services provided. A pilot study was initiated in which individuals who received genetic counseling for cancer were asked to provide feedback on the perceived quality of the services provided, and to identify areas in which additional services may be required. Preliminary results based on 36 counseled individuals indicated generally high levels of satisfaction with the care provided by the clinical geneticist and with the procedures at the familial cancer clinics. Several areas were identified that deserve additional attention: (1) the role of the family doctor in the genetic counseling; (2) communication of information regarding the possible impact of genetic counseling and testing on daily life; (3) communication between the clinical geneticist and other health care workers, and (4) psychosocial support during and after the process of genetic counseling. PMID- 9355579 TI - A review on family history of breast cancer: screening and counseling proposals for women with familial (non-hereditary) breast cancer. AB - With the aim to specify screening recommendations for women with familial (non hereditary) breast cancer (FBC) we analysed 59 studies using quantitative methods of pooling. The pooled relative risk (RR) and cumulative probability were used to estimate breast cancer risk. The RRs for women with a family history of breast cancer in a first-degree relative was 2.03 (95% CI 1.09-2.22). The highest RR is observed for women with a family history and atypical hyperplasia in their breast biopsy specimen (RR = 10.87, 95% CI 6.05-19.69). A high cumulative probability before the age of 50 was only found for women with a combination of two risk factors: a family history and atypical hyperplasia, namely 19% (95% CI 11-33%). The cumulative probabilities of women aged 50 to 70 years who have a family history were between 11% (95% CI 9-13%, a family history in combination with age at first birth before 22 years) and 53% (95% CI 35-75%, a family history in combination with atypical hyperplasia). These high risks suggest that women over 50 years of age who have a family history of FBC have to be actively encouraged to participate in a screening program consisting of a biannual palpation by a specialist, an annual mammogram and a monthly self-control. Yearly screening is recommended for women under 50 years of age who have a family history and atypical hyperplasia. These recommendations remain valid until the effectiveness of such screening programs is assessed. PMID- 9355580 TI - Uncertainty in the information provided during genetic counseling. AB - Clients seek genetic counseling in order to become informed, to make better decisions, and, if possible, to be reassured. Genetic knowledge, however, is fragmentary and incomplete and therefore it may involve more uncertainty than is desirable. In a cohort of 30 counseling sessions we studied the genetic information that was actually conveyed in terms of its predictability, controllability and novelty. With regard to predictability it emerged to be rather the rule than the exception that clients of genetic counseling were confronted with (1) an inconclusive diagnosis, (2) the chance or an estimate of the chance of the occurrence or recurrence of a genetic disorder, and (3) ambiguity about the severity of the disease. In case of bad news, possibilities for control (therapeutic or preventive measures) were minimal. In a few cases, clients were confronted with completely unexpected findings, i.e., information of high novelty. It is concluded that the high degree of uncertainty in the information provided during genetic counseling--reflecting the true state of the art--is in direct contrast to the needs of clients. PMID- 9355581 TI - Epilepsy surgery for partial seizures. AB - Epilepsy, a common neurological condition most often diagnosed in childhood, has physical, psychologic, and psychosocial effects on individuals. Of people diagnosed with complex partial seizures, about 10%-20% will not be able to achieve control of their seizures with medical management. The impact of continued seizures can be devastating on developing children and their families. Surgery for removal of the epileptic focus can offer the hope of an ordinary life; over 70% of patients achieve either complete seizure control or are greatly improved (Ventureyra & Higgins, 1993). Surgery requires a careful evaluation, services of a multidisciplinary team, and education and support for children and families. Pediatric nurses can identify potential candidates, act as advocates to encourage families to seek this option, and work collaboratively with their colleagues in specialty centers to facilitate referrals. PMID- 9355582 TI - Use of the ketogenic diet in treating children with seizures. AB - Ketogenic diet therapy has been found to be an effective means of treating afebrile seizures that are refractory to antiepileptic medication alone. Controversies exist regarding its use. Potential harmful side effects include Staphylococcus aureus infections, retarded growth, hypoglycemia, hyperlipidemia, urolithiasis, and optic neuropathy. Pediatric nurses with knowledge about ketogenic diet therapy and current research regarding its use, will be better able to determine the appropriateness of this form of therapy for children with seizures that cannot be controlled by medication alone. PMID- 9355583 TI - An interdisciplinary team approach to implementing the ketogenic diet for the treatment of seizures. AB - The ketogenic diet is becoming a more recognized method of treating seizures in some children with epilepsy. Texas Scottish Rite Hospital for Children (TSRHC) developed an interdisciplinary team that uses a comprehensive approach to implementing this innovative therapy. Anticipated outcomes of this unique approach are increased family satisfaction and improved dietary compliance, which maximizes the diet to its fullest potential. These outcomes are validated by anecdotal information by families' reports of satisfaction and successful diet maintenance for increased number of months (this population from 3-31 months). Of the 27 children ages 1-16 years, approximately 40% have experienced reduction of seizures of more than 50%, 25% are seizure free, and 35% have discontinued the diet for a variety of reasons including difficulty in consistently maintaining the diet. Originally introduced in the 1920s, the ketogenic diet has once again become cutting edge treatment for nonresponsive seizure activity. PMID- 9355584 TI - Heliox: a new treatment for life-threatening asthma. AB - Rates of asthma are increasing in adults and children. In some pediatric institutions, asthma is the most common admitting diagnosis. At University of Chicago Children's Hospital (UCCH), at the University of Chicago Hospitals, the use of helium-oxygen mixtures (heliox) has been explored for treatment of children in status asthmaticus. Helium has a low density, and therefore has an increased flow rate which results in less airway resistance, decreased work of breathing, minimized airway collapse, and less hyperinflation. Little research on heliox and asthma has been done with children. A small study of children with status asthmaticus at the University of Chicago Children's Hospital found heliox resulted in improved peak flow readings and reductions in dyspnea and pulsus paradoxus. Nursing care of the child receiving heliox includes the use of appropriate delivery devices; monitoring temperature and other potential side effects; educating the child and family; and regularly assessing the following: pulse oximetry readings, heart and respiratory rates, blood pressure, and pulsus paradoxus. PMID- 9355585 TI - Reliability of head circumference measurements in preterm infants. AB - PURPOSE: To describe and compare the intra- and interexaminer reliability of head circumference measurements obtained with paper and cloth tape measures. METHOD: Two experienced neonatal nurses each obtained head circumference measurements using both paper and cloth tape measures twice each, from 49 clinically stable, preterm infants. The nurses were blind to their own and to each other's measurements. The order in which the measurements were obtained was randomized. The differences within and between examiners for cloth and paper tape measurements were described using mean absolute differences, standard deviation of net differences, technical error of measurement, minimal and maximal differences, percentage of differences 0.25 and 0.5 cm, and percentage of error. RESULTS: Wilcoxon matched-pairs, signed-ranks tests demonstrated significantly greater intraexaminer reliability for measurements obtained with the paper tape for both of the nurses. Wilcoxon matched-pairs, signed-ranks tests also demonstrated significantly greater interexaminer reliability for measurements obtained with the paper tape for both the first and second measurement sets. CONCLUSIONS: Intra- and interexaminer differences were consistently smaller when the examiners used paper tape measures. PMID- 9355586 TI - Creating a beginning ethics library. PMID- 9355587 TI - Pediatric Nursing Humanitarian Award recipient: Rear Admiral Julia R. Plotnick. PMID- 9355588 TI - Books for siblings of children having illness or disability. AB - A child's disability or acute or chronic illness has an effect on all family members, including siblings of the affected child. Children's books, fiction and non-fiction, can be used to support siblings by providing information, addressing feelings, offering insight, assuring the child that others have had similar problems, and facilitating coping. Nurses can introduce children and their families to useful books by having books or book covers on display that the child can choose from; providing families with lists of books; and/or assessing the child's needs, interests and reading level, and recommending a specific title. PMID- 9355589 TI - Fosphenytoin sodium: new drug to replace intravenous phenytoin sodium. PMID- 9355590 TI - School services for children with chronic conditions. AB - School is critically important for all children, including those with chronic conditions. Yet school services are often difficult for these children to obtain. This article provides an overview of chronic conditions, a description of the relevant laws and their service provisions, and a discussion of ways nurses can facilitate children receiving specialized education. PMID- 9355592 TI - Preventing insurance denials: disease management. PMID- 9355593 TI - Balanced budget act of 1997 enacted in August includes child health initiative. PMID- 9355591 TI - Pediatric management problems--Kawasaki disease. PMID- 9355594 TI - The use of heparin and normal saline flushes in neonatal intravenous catheters. AB - PURPOSE: To compare the effects of heparin versus normal saline flush solutions on the duration of patency of peripheral IV catheters in neonates. METHODS: A quasi experimental design compared the outcomes in 87 infants who were 32 weeks or greater gestation at birth. Thirty-three received heparin and 54 received normal saline. RESULTS: There were 159 starts among the subjects. No statistically significant difference in duration of patency between the groups was found. The duration was significantly longer for term than preterm infants and for insertion in scalp, arm, or hand than for leg or foot. CONCLUSIONS: Gestational age and site of insertion were the only predictor variables related to duration of patency for IV catheters. PMID- 9355595 TI - Leadership principles: lessons learned. PMID- 9355597 TI - Pittsburgh nurse works with teens to prevent pregnancy. Interview by Elizabeth Todd. PMID- 9355596 TI - Institutional policies on the use of physical restraints on children. AB - Within health care institutions, nursing policies often serve as the gold standard for nursing practice. Policies regarding the use of physical restraints on children are typically not based on any scientific evidence. This article analyzed multiple hospital policies and makes recommendations to better assess whether or not restraints are needed, suggests a list of least to most restrictive devices, identifies interventions for the child in restraints, and offers alternatives to the use of restraints. PMID- 9355598 TI - The resume: minimizing the impact of a demotion. PMID- 9355599 TI - Enteral nutrition: hospital to home a collaborative approach to education and care. PMID- 9355600 TI - Increasing the effectiveness of your teaching program for the elderly: assessing the client's readiness to learn. AB - The education of the elderly person is a critical component of nursing interventions for health maintenance and illness prevention in the elderly. Teaching and learning transactions require great skill under the best of circumstances, but they take on new demands and require different skills when the client is ill and feels vulnerable. To maximize an educational opportunity, the nurse must be able to understand the factors that may inhibit successful learning and be very astute in assessing and re-evaluating the client's readiness and capacity to learn at a specific time. It is imperative that nurses realize that taking the time to assess a client's readiness to learn and utilizing the information that is gathered to develop a teaching plan that best meets the clients needs increase the likelihood of a successful teaching/learning event for both the nurse and the client. The information should assist the health care worker's ability to give older persons the tools and information that they require to assist them to maintain optimum independence and a better quality of life. Hopefully, the end result will be a win-win encounter for all involved in the learning experience. PMID- 9355601 TI - Tee-up garden: aggression redirected. PMID- 9355602 TI - Developing the perfect long-term mattress. PMID- 9355603 TI - Science by consensus. PMID- 9355604 TI - Ombudsman: a new role for advanced practice nurses in psychiatric-mental health nursing. AB - TOPIC: The role of ombudsman for advanced practice nurses in psychiatric mental health nursing. PURPOSE: To describe the role of ombudsman and its fit with nursing as seen in the Price Spratlen Ombudsing Model. SOURCE: The author's own experiences as both an advanced practice nurse and an ombudsman. CONCLUSION: Because of downsizing, reorganization, and a general trend toward mutual distrust in large organizations, being an ombudsman has been named one of the "25 hottest careers." Advanced practice nurses in psychiatric mental health nursing, by virtue of their knowledge of interpersonal, preventive, and systems theories, are in a unique position to fill this role. PMID- 9355605 TI - Resolution of drinking problems without formal treatment. AB - TOPIC: Self-resolution of alcohol abuse among individuals with mild to moderate drinking problems. PURPOSE: To examine self-resolution of drinking problems and clinical/research implications. SOURCE: Review of the literature. CONCLUSION: Two pathways toward self-resolution of alcohol problems have been proposed. One appears to be relatively spontaneous, the other involves ongoing cognitive appraisals and a conscious effort to change. Nurses have the potential to play an important role in encouraging self-change by conducting screenings, implementing brief interventions, and promoting the use of self-help materials. Opportunities also exist for nurses to spearhead research efforts in this long-neglected area. PMID- 9355606 TI - The psychotherapy of Hildegard Peplau in the treatment of people with serious mental illness. AB - TOPIC: The use of Peplau's interpersonal nursing theory with people suffering from serious mental disorder. PURPOSE: To describe Peplau's theory and its application using a case study. SOURCE: Author's own clinical work. CONCLUSION: Peplau's theory can be used to help patients resolve symptoms by guiding them through the steps of observation, description, analysis, formulation, validation, testing, integration, utilization. PMID- 9355608 TI - Nurse in "catch-22" situation: court reverses disciplinary action. PMID- 9355607 TI - Loss of control and the chronic mentally ill in a rural day-treatment center. AB - PROBLEM: What do chronically mentally ill clients consider to be the core variable in maintaining health, and how do they go about doing this? METHODS: A grounded theory design with analysis of interviews of seven chronically mentally ill clients in a rural day-treatment center. FINDINGS: The core variable in maintaining health according to chronic mentally ill clients is preventing loss of control. A process was discovered that clients used to do this. CONCLUSIONS: Chronic mentally ill clients prevent loss of control by using informal relationships to adapt behaviors, attitudes, and feelings within a supportive environment. If this fails, they turn to formal sources of control such as therapists, case workers, or other mental health providers. PMID- 9355609 TI - Can "volunteers" invoke the Good Samaritan rule? Case on point: Boccasile v. Cajun Music Ltd. 694 A. 2d 686--RI (1997). PMID- 9355610 TI - NY: Nurse's aide barricades pt's door: "mistreatment of patient" charge affirmed. FL: aide charged with neglect of aged pt.: finding of neglect reversed-record expunged. PMID- 9355611 TI - Nurse alleges Dr. mistreated "black children": retaliatory termination. Case on point: estate of Parks v. O'Young 682 N.E. 2d 466--IL (1997). PMID- 9355612 TI - AASCIN: embracing a new tomorrow. PMID- 9355613 TI - Spinal cord injury nurses in action: partners in practice. AB - The creation and implementation of a Partners in Practice care delivery model on a spinal cord injury unit is described in this article. As a result of changes in the number and mix of staff, new strategies were necessary to assure the provision of patient focused, team oriented care. The goal was to increase the efficiency, effectiveness, and continuity of care while ensuring staff accountability and appropriate utilization of resources. Manthey's (1980) Partners in Practice model was adapted to meet the needs of the unit, and was implemented using Lewin's (Scott, 1994) change theory. Outcomes of the model included the establishment of a culture of partnership for providing team oriented care to spinal cord injury patients. PMID- 9355614 TI - Managing autonomic dysreflexia through the use of clinical practice guidelines. AB - One of the goals listed in the American Association of Spinal Cord Injury Nurses (AASCIN) strategic plan 1993-1998, is to provide strategic vision to advance professional practice in spinal cord injury (SCI) nursing. In this quest, along with the development of Standards of Spinal Cord Injury Nursing Practice, the AASCIN Clinical Practice Committee is developing Clinical Practice Guidelines to describe a suggested course of action to address specific clinical conditions or needs of individuals with spinal cord injury. The first Clinical Practice Guideline to be completed is Autonomic Dysreflexia (AD) (Kuric & Hixon, 1996). The purpose of this article is to review the pathophysiology of AD and then describe the symptoms, assessment parameters and nursing interventions suggested in the Clinical Practice Guidelines. PMID- 9355615 TI - Knowledge acquisition and decision-making: spinal cord injured individuals perceptions of caring during rehabilitation. AB - Nurses and other healthcare providers have little research to guide them on specific interventions and attitudes which expedite the attainment of rehabilitation outcomes by spinal cord injured (SCI) individuals and their successful return to the community (Fuhrer, 1994; Whiteneck, 1994). Acquisition of knowledge is required during rehabilitation to learn self-care and decision making which is essential to long term survival following SCI. However, skills that patients and families are able to accomplish in rehabilitation are often not able to be translated into the home environment (White & Holloway, 1990). The process of learning self-care and decision-making needs to be more clearly elucidated, so more effective interventions can be designed which can improve problem-solving and lead to enhanced well-being and quality of life. The purpose of this qualitative study was to describe the meaning, process and consequences of caring during rehabilitation from the perspective of the SCI individual. This paper will report on the findings from one research question: How is the process of a developing caring relationship perceived by SCI individuals during rehabilitation? The theoretical foundation of caring for this study was synthesized from philosophical, ethical, feminist, and nursing literature. A purposive sample of adults with traumatic SCI were interviewed at least once during their initial rehabilitation stay. Twenty interviews were conducted with fifteen participants at various times during their rehabilitation stay over a six month period. The core category of "getting back together" or reintegration of self, which was the major work of rehabilitation, was accomplished with nurses and therapists who were perceived as caring. The process of a developing caring relationship was conceptualized, from participants' descriptions in three phases: learning the other, learning what I need to know, and letting me find out. During each phase reciprocal behaviors occurred between the patient and the caregiver resulting in intermediate outcomes, which facilitated movement to the next phase. Consequences of these caring relationships were: well-being, self-care, autonomy, independence, and hope. Caring by rehabilitation professionals was perceived by SCI individuals as central to recovery and to a positive attitude toward disability. PMID- 9355616 TI - How can nurses use limit setting to facilitate spinal cord patients' independence? AB - Learning limit setting techniques is necessary for the entire team in order to have consistent positive patient outcomes. Team building exercises will assist in supporting restructuring of acute care units' healthcare delivery. Difficult patients need to participate in their plan of care along with representatives from each discipline. Caregiver competency in psychological, physical, technical, and physiological aspects of care for the SCI patient facilitates patient independence. This is a result of our increased knowledge which teaches us that these kind of patients can be more independent and, hence more in control, than caregivers may realize. Patients will be less likely to be manipulative and abusive to the healthcare staff if they feel more in control of their care and their lives. PMID- 9355617 TI - [With Marianne in home care nursing. Interview by Susanne Bloch Kjeldsen]. PMID- 9355618 TI - [With Maja-Lis on the medical department. Interview by Charlotte Frendved Hansen]. PMID- 9355619 TI - [With Lisbeth in health visiting. Interview by Dorthe Nerving]. PMID- 9355620 TI - [With Vibeke on the intensive care department. Interview by Kirsten Bjornsson]. PMID- 9355621 TI - [Better wage and working conditions for health visitors]. PMID- 9355622 TI - [Psychiatry--a super section for young people]. PMID- 9355624 TI - [Drug dependence--Denmark lacks treatment facilities]. PMID- 9355623 TI - [Clinical competence development in the health system]. PMID- 9355625 TI - [Sweden: drug dependence--treatment takes time]. PMID- 9355626 TI - [Drug dependence--suffering caused by the health system. Interview by Charlotte Frendved Hansen]. PMID- 9355627 TI - [Drug dependence--increasing abuse of pain-killing medicine]. PMID- 9355628 TI - [Executive Board--strategy for new wage forms]. PMID- 9355630 TI - [Habits control our opinion about patients]. PMID- 9355629 TI - [Merit students can fulfill needs]. PMID- 9355631 TI - [Vulnerable young mothers]. PMID- 9355633 TI - Unsung nursing heroes. PMID- 9355632 TI - Legislative victories. PMID- 9355634 TI - Communicating with older adults. PMID- 9355635 TI - [Violence against nurses]. PMID- 9355637 TI - [Education in operating room assistance has started]. PMID- 9355636 TI - [Few concrete things in election campaign. Interview by Marit Fonn]. PMID- 9355638 TI - [A working day with harassment, threats and violence]. PMID- 9355639 TI - [Viewpoint from audiometry. Interview by Marit Fonn]. PMID- 9355640 TI - [Cognitive insufficiency--the invisible functional inhibition. Interview by Kjell Arne Bakke]. PMID- 9355641 TI - [Close-up: Karen Bjoro, newly appointed chief nurse in Ulleval Hospital--from expert to novice. Interview by Kjell Arne Bakke]. PMID- 9355642 TI - [Recruitment--untraditional hunt for nurse specialists]. PMID- 9355643 TI - [My working place: Rikshospitalet, Oslo--coordination of transplantations. Interview by Kari Anne Aase]. PMID- 9355644 TI - [From times gone by--treatment of bed sores]. PMID- 9355645 TI - [Vaccination--an effective infection protection initiative]. PMID- 9355646 TI - [ICN Congress--trends and policies in health services]. PMID- 9355647 TI - [Staffing--poor nursing care in the holidays. Interview by Kjell Arne Bakke]. PMID- 9355648 TI - [Multi-cultural Norway--elderly immigrants will not be in the pension structure. Interview by Marit Fonn]. PMID- 9355650 TI - [Forced medication in a somatic nursing home]. PMID- 9355649 TI - [Uganda--a visit to the hospital and nursing school in Masaka]. PMID- 9355651 TI - [Professional Ethics Council--family's decision making authority and comments to the media]. PMID- 9355652 TI - [Handling conflict--it is disconcerting to be between the devil and the deep blue sea]. PMID- 9355653 TI - [Ethics--sympathy, action and priority setting]. PMID- 9355654 TI - [Documentation--classification of nursing procedures]. PMID- 9355655 TI - [Psycology, behaviorism and nursing care]. PMID- 9355656 TI - [Violation of integrity in compulsory mental health protection]. PMID- 9355657 TI - Biochemical screening of congenital developmental abnormalities using determination of fetoplacental antigens. AB - In a prospective study of screening of maternal serum levels of alpha-1 fetoprotein (MS AFP) and trofoblast-specific beta-1-glycoprotein (MS SP1) were examined in samples from pregnant women between the 16th and 18th week of pregnancy. We detected 8 fetuses with chromosomal aberation, 8 fetuses with neural tube defects and 10 fetuses with inborn cardial defects. Our study confirms higher MS SP1 levels in women with fetuses with chromosomal aberation, while MS AFP's tendency is to decrease. When combining MS SP1 + MS AFP + age of mother (over 35), 75% of fetuses with chromosomal aberation were detected. In women with neural tube defect 75% fetuses were detected by MS AFP + MS SP1 combination. PMID- 9355658 TI - On the interpretation of "mean +/- s", II. Alternative questions and answers. PMID- 9355660 TI - Prolonged oral glucose tolerance test combined with indirect calorimetry and estimation of several substrates and hormones in obese subjects. AB - OGTT prolonged to 4 hours was performed in untreated 50 obese outpatients (mean age 37.9 years and BMI 37.2). Parallel to glucose, insulin, growth hormone (GH), cortisol, beta-hydroxybutyrate (BOHB), nonesterified fatty acids (NEFA) and indirect calorimetry (RQ) were estimated at hourly intervals. Basal values of thyroxine (T4), triiodothyronine (T3), total cholesterol and triacylglycerols were also obtained. Mean glycemia after overnight fasting as well as after 75 g glucose reached upper limits of the normal range. The subsequent decrease was slower. Insulinemia followed a similar trend. The initial drop of GH after the glucose load reverted to an increase above the basal value. A similar pattern was observed with cortisolemia, but the decrease and increase were less important. The basal value of RQ was rather low and glucose ingestion produced only a small increase, followed by a greater decrease. Serum levels of NEFA and BOHB sharply decreased one hour after glucose intake and afterwards regained the initial values. The mean basal values of T4 and T3 were within the normal range--the low T3 syndrome was not involved in the large majority of cases. Cholesterol and triacylglycerols approached the upper normal limit. The correlations brought additional information. Insulinemia increased parallel with the amount of body fat. The basal level was decisive for most hormones and substrates--the high or low set level could be followed in the course of the whole test. Increased insulinemia and increased glycemia suggested the presence of a mild insulin resistance with the participation of GH and cortisol. Increased levels of fasting insulinemia and glycemia were present also in obese subjects with a normal OGTT. The correlations permitted to disclose insulin-like effects of GH on basal conditions. Increased BOHB was responsible for a high cholesterol. It is suggested that even small fluctuations of glycemia related to food intake may produce a substantial modification of the hormonal status in obese subjects and initiate or support the metabolic disorders in obesity. In this respect a greater role is ascribed to the phase of decreasing glycemia in comparison to the increasing phase. Lipids are the prevailing source of energy in insulin resistant subjects. The rather stable values of indirect calorimetry indicate that energy metabolism of obese subject works on a low, pre-set level-independently on the supply of some relevant hormones and substrates. PMID- 9355659 TI - Hormonal and metabolic adaptation to a reducing regimen in children. AB - The effect of a combined slimming regimen (5.2 MJ diet and a low to medium intensity motor activity) were studied in 20 moderately obese children. Blood samples were obtained on days 0, 10 and 52 of the regimen. Insulin and triacylglycerols decreased after 10 days of treatment and the reached levels were maintained up to day 52. Cortisol, T4 and T3 decreased throughout the whole regimen. Glucose and lactate first decreased, and then increased to reach initial levels. NEFA and beta-hydroxybutyrate moved opposite to glucose. The initial decrease and the terminal very high increase of growth hormone were, however, statistically not significant. The serum proteins remained unaffected. In addition to a mean loss of 9 kg, the favourable effect of the slimming regimen consisted in the decrease of insulinemia, cortisolemia and triacylglycerolemia. The unfavourable effect was seen in the decrease of T3, responsible for the decreasing weight loss in the course of slimming regimens. PMID- 9355662 TI - Effects of some factors on the indication for surgical treatment of gastroduodenal ulcer disease. AB - The author of the present communication collected a variety of important information on 2,173 patients living permanently in Prague 4. In all of them the diagnosis of gastroduodenal ulcer disease was established between 1979 and 1988 incl. The results obtained by the currently applied therapy were not designated as satisfactory both by the patient and by his doctor. The predomination of conservative therapy was due to the introduction of new drugs which reduce markedly gastric secretion of HCl. On the basis of strict indications elective surgery was advised in some patients. In the acute stage of the disease (bleeding, perforation, stenosis) clearly defined criteria allowed to establish the indication for elective surgery. The answer to this question provides the author in the form of a scheme of the regimen which should be observed after a definite diagnosis. It was devised on the basis of the results of the present study, of reports in the Czech and international literature, as well as of our own experience. For the indication of surgical treatment is of great importance a high degree of responsibility of the doctor. This is reflected by the correct choice of the optimum surgical procedure. Good results obtained in the management of ulcer disease were due to the fact that attention was paid to the often neglected aspect of the psychosomatic character of this disease. PMID- 9355661 TI - An application of neutron activation and radiotracer techniques for studying the status of sodium in the bone mineral. AB - Neutron activation analysis of intact and water-extracted (on boiling) bone (femur) samples made it possible to determine manganese, sodium, potassium and chlorine. A major portion of sodium in contrast to potassium) was retained in the bone, which was in agreement with a well known presence of slowly exchangeable sodium in the bone. Further experiments were carried out with fragments of the femur of rabbits that were administered with the radionuclide 22Na and sacrificed after a sufficient equilibration period. Under conditions of stirring with water at a laboratory temperature, a two-exponential course of the sodium ion release was observed with half time values of 4.36 and 167 hours. An explanation is suggested that in the bone there are actually two fractions of slowly exchangeable sodium, present on the bone mineral crystal surface and bone mineral crystal interior, respectively. The experiments on the bone in vitro can help to bridge a gap between in vitro experiments on synthetic hydroxy apatite and dynamic studies based on the in vivo neutron activation analysis. Results of a complementary experiment with ashed bone samples demonstrated an almost complete abolishment of the sodium ion release, which is in perfect agreement with a known concept of the thermal recrystallization of the bone mineral. PMID- 9355663 TI - Bone mass during growth: the effects of exercise. Exercise and mineral accrual. AB - Intense exercise during childhood and adolescence may result in primary amenorrhea and low peak bone mineral density (BMD). After puberty, exercise may result in secondary amenorrhea and bone loss. Higher BMD in amenorrheic athletes than amenorrheic sedentary persons suggests that exercise may partly offsets the effects of amenorrhea. To examine this possibility, we measured BMD (g/cm2) by dual x-ray absorptiometry in 32 ballet dancer and 23 healthy controls of comparable age with regular menstrual cycles, 34 pre-pubertal female gymnasts bone age 8.9 +/- 0.2 years and 37 girls matched by bone age. Dancers had normal BMD at the weight bearing sites, not low, despite having oligomenorrhea, not high despite 32 hours of week dancing. BMD was lower by 4-6 percent at the non-weight bearing sites. BMD diminished in the dancers at the weight bearing femoral neck (r = -0.29, P = 0.1) and trochanter (r = -0.31, P = 0.09), and at the non-weight bearing arms (r = -0.29, P = 0.09) with increasing duration of amenorrhea. Dancers with less than 40 months amenorrhea had 5 to 7% higher BMD at the weight bearing, but not non-weight bearing sites. Dancers with more than 40 months amenorrhea had normal, not higher BMD at weight bearing sites and deficits of about 5 percent at non-weight bearing sites. In gymnasts, BMD was 10-15 percent (or 1 SD) higher than the bone age-predicted mean. Exercise may not offset the effects of amenorrhea. Bone loss may continue but from a higher level, perhaps attained prior puberty. PMID- 9355664 TI - Determinants of bone growth and strength in juvenile (rheumatoid) arthritis. Interaction with other influences. PMID- 9355665 TI - Risk factors for osteoporosis and densitometric surveillance in children with chronic rheumatoid diseases. PMID- 9355666 TI - Monitoring bone mineralization in chronic rheumatic children on steroid treatment. PMID- 9355667 TI - Potential of densitometric measurement in selected pediatric subject. AB - On the aim to recognize bone mineral measurements as somatic development parameter as well as its clinical usefulness in pediatric clinic, the investigations with of DPX-L were performed. Material consisted of 302 healthy scholars both sex and 85 patients aged 6.0-18.9 yrs children diagnosed as idiopathic juvenile osteoporosis (IJO) and osteogenesis imperfecta (OI). Total body mineral density (TBBMD) was evaluated as developmental parameter in normal children population and utilized in differential diagnosis and monitoring of bone pathology in children with IJO and OI. PMID- 9355668 TI - Bisphosphonates: pharmacology and effects on the growing skeleton. PMID- 9355669 TI - Juvenile chronic arthritis--bone mineral density in relation to corticosteroid therapy. AB - Bone demineralization often accompanies juvenile chronic arthritis (JCA). Fourteen patients with confirmed diagnosis of JCA had their bone mineral density (BMD) measured with the use of dual photon X-ray absorptiometry. The results obtained were compared to the Lunar BMD DPXA standards. Seven patients received Prednisone in doses of more than 0.16 mg/kg/day for more than 6 months and 7 patients (sex and age matched) never received any steroids at all. In the first group BMD was decreased in 7 patients (100%), in the second group BMD was mildly decreased in 2 patients (28%). Due to the difference in BMD in both groups, it is obvious that corticosteroids have substantial influence on bone demineralization in JCA. PMID- 9355670 TI - Investigations of bone density in patients with juvenile chronic arthritis by a sonographie method. PMID- 9355671 TI - Calciuria in children with juvenile chronic arthritis. AB - Demineralization of bone is a frequent finding in children with juvenile chronic arthritis (JCA). Recently there have been reports about hypercalciuria accompanying JCA. This is believed to be associated with increased bone resorption due to cytokines and immobility of the patients and steroid treatment. In 12 patients with confirmed diagnosis of JCA basic biochemical indices of bone metabolism, were performed (S-Ca, P, ALP, U-Ca/U-creatinine, U-P). Bone mineral density (BMD) was measured using DPXA method and results obtained were compared to the Lunar BMD DPXA standards. In spite of decreased BMD, no significant hypercalciuria was found and other mentioned biochemical indices of bone and mineral metabolism were normal as well. PMID- 9355672 TI - Bone and mineral metabolism in transient hyperphosphatasaemia. AB - Transient hyperphosphatasaemia (TH) is a benign disorder characterized by transient elevation of S-ALP activity not exceeding duration of 4 months in children under 5 years of age, with elevated activity of bone isoenzymes of ALP with no signs of bone or liver disease and variable unrelated symptoms. We observed 19 children with TH and in 3 patients with markedly elevated S-ALP activity we found increased excretion of urinary hydroxyproline, suggesting increased bone resorption followed by bone formation. In 3 children with history of TH, bone mineral density (BMD) was measured and found to be normal. Transient increased bone resorption followed by bone formation during the course of TH can not be ruled out, but this has no negative impact on BMD. PMID- 9355673 TI - Two cases of pseudohypoparathyroidism in adolescent boys. AB - Pseudohypoparathyroidism (PHP) is characterized by end organ resistance to parathyroid hormone (PTH). PHP type Ia consists of Albright's osteodystrophy and resistance to PTH. In PHP type Ib physical appearance is normal and there is no response to PTH in U-cAMP excretion. In PHP type II both physical appearance and U-cAMP response to PTH infusion are normal. Two adolescent patients with severe hypocalcaemia were treated in our department. The first boy was admitted because of low back pain, latent tetany and recurrent collapsing, the second one due to Grand mal epilepsia. S-Ca concentrations were very low (1.1 and 1.03 mmol/l respectively), CT of the brain revealed multiple calcifications in basal ganglia and S-PTH concentrations were above upper reference level. Therefore the diagnosis of PHP was established. In the absence of skeletal malformations the most probable diagnosis is PHP Ib or II. Clinical state of the boys has dramatically improved after calcium and vitamin D supplementation. PMID- 9355674 TI - The importance of milk in nutrition and development of its consumption. AB - The author describes the development of milk consumption in CR since 1989 and its downward trend during the course of transformation period. He refers to the importance of milk proteins content and especially of calcium in dairy products. He compares the level of milk consumption in CR, in terms of milk proteins and milk fat content, with selected countries with developed dairying. He assumes that the drop of the average milk and dairy products consumption in the period 1989-1990 caused 140 mg reduction of daily calcium intake per 1 inhabitant. PMID- 9355675 TI - Metabolic bone status in young women with JCA. AB - The study was based on clinical, densitometric and biochemical evaluations and on a life-style questionnaire, applied in a cohort of 41 individuals. The mean age of the young women was 24.2 (18-36) years. The main point was to compare subgroups with (CS) an without (NCS) corticosteroid treatment (19 and 22 patients, respectively). There was no significant difference in age, weight and duration of JCA. Of the densitometric examinations, spine DXA (DPX-L LUNAR apparatus) yielded values significantly lower in CS than in NCS individuals (p = 0.05). Much more apparent was the difference in stiffness measurements in the calcanei performed by Achilles-LUNAR ultrasound instrument, again with lower values in CS women (p = 0.001) (Fig. 1, 2, 3). No significant differences were found between the two subgroups as regards blood levels of bone alkaline phosphatase, osteocalcin and tartrate-resistant acidic phosphatase, and urine hydroxyproline output. Menarche occurred at a mean age of 14.37 years in the CS subgroup (p = 0.01, against healthy population) and of 13.32 years in the NCS subgroup (not significant). The prevalence of fractures was enconsiderable in both subgroups. These findings are to be understood from the viewpoint of combined influences of both disease activity and corticotherapy in the CS patients with JCA. PMID- 9355676 TI - HLA and juvenile rheumatoid arthritis. AB - HLA class II analysis in a group of 153 Czech children with juvenile rheumatoid arthritis by PCR and oligonucleotide hybridization demonstrated associations with several alleles. DRB1*0801 (RR = 5.3, p < 0.005) and DRB1 * 11 (RR = 2.2, p < 0.01) including all subtypes were shown to be increased in the rheumatoid factor negative group (N = 137). The same results were observed in Italy, England and Norway. In patients with the pauciarticular onset with conversion to polyarticular within 3 years, a statistically significant increase in DR2 (RR = 10.1, p < 0.00005), mostly due to DRB1*1501, was found. In the iridocyclitis and antinuclear factor groups, susceptibility to DRB1*1201 was observed. There was a striking decrease in DRB1*0701 (RR = 0.3, p < 0.00005) in all groups. There was neither an increase in DRB1*1301 or DPB1*0301 nor a decrease in DRB1*04, as reported from other studies in Texas and Norway. The rheumatoid factor-positive group with polyarticular onset (N = 13) was associated with DRB1*04 (RR = 7.1, p < 0.005), as observed in adults. DPB1*0201 was increased in the persistent pauciarticular group (RR = 3.7, p < 0.0005). DPB1*0402 was decreased in all pauciarticular groups with or without conversion (RR = 0.3, p < 0.005). Taken together, there are not only genetic differences and clinical heterogeneity in juvenile rheumatoid arthritis patients but, also, common predisposing factors. PMID- 9355677 TI - Chemiluminescence of peripheral blood leukocytes and activity of an inflammatory process in juvenile chronic arthritis (JCA). AB - The oxidative metabolites have been implicated in the aetiology and pathology of rheumatoid arthritis. In our work we endeavoured to deal with polymorphonuclear leukocytes (PMNLs) ability to generate oxygen free radicals (OFRs). PMNLs metabolic activity was assessed by the means of chemiluminescence (CL) method. We observed significantly higher PMNLs metabolic activity in children suffering from JCA in comparison to the healthy group. Greater activity of the rheumatoid process was accompanied by an increased PMNLs activity. The results constitute another piece of evidence confirming the role of PMNLs in the pathogenesis of JCA. PMID- 9355678 TI - Levels of circulating intercellular adhesion molecule 1 and E-selectin in serum and synovial fluid of juvenile chronic arthritis patients. AB - OBJECTIVE: To measure the levels of two adhesion molecules (AM), soluble intercellular adhesion molecule 1 (sICAM-1) and soluble E-selectin (sE-selectin), in serum and synovial fluid (SF) of patients with juvenile chronic arthritis (JCA). METHODS: Both soluble AM levels were tested, in serum and synovial fluid (SF) samples, with an enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Serum levels of sICAM-1 and sE-selectin in JCA patients were not significantly different from those of a control group. Synovial fluid levels of sICAM-1, but not of sE-selectin, assayed significantly higher (p < 0.05) in JCA patients than in controls. Moreover SF levels of both molecules correlated negatively with disease duration, being higher in the earliest phases. No significant correlations were found between JCA sICAM-1 and sE-selectin levels and leukocyte count or ESR. CONCLUSIONS: These observations may signify a more important role of ICAM-1 than E-selectin in the migration of inflammatory cells into JCA SF. The negative correlation of both AMSF levels in JCA patients with disease duration could reflect a higher expression of ICAM-1 and E-selectin during the earliest phases of the disease. PMID- 9355679 TI - The production of saliva of patients with juvenile chronic arthritis (JCA). AB - About 12-16 p.c. of the patients with JCA had a reduced saliva production. These were older than twelve years or belonged to the systemic form of JCA. The reduced saliva production is clinically not discernible. Enteral deviation or disturbances are observed with rheumatic arthritis. Various examinators have found a Sicca syndrome at about 6-12 p.c. of patients suffering from rheumatic arthritis (1, 2, 3). The mostly retrospective examinations do not allow a direct comparison. Up to now the Sicca syndrome has rarely been observed with the Juvenile Chronic Arthritis. This investigation is to present the quantitative production of saliva of JCA patients. PMID- 9355680 TI - Some remarks on management of juvenile chronic arthritis complicated by amyloidosis. AB - A retrospective study of 66 juvenile arthritic patients with reactive amyloidosis for a mean 12 +/- 10.7 years (1 to 30 years) from the onset of amyloidosis was performed. Forty-seven of them received continuous long-term chlorambucil therapy. Nineteen remaining patients were not treated with cytostatics of less than 3 months. The mortality rate for the whole group was 54.5%. 63.8% of those treated with chlorambucil are alive (mean survival 23.1 +/- 2.5 years). All patients from untreated group died. Their mean survival was 3.2 +/- 2.2 years. Nine patients from another group had been given thymus peptides for six months. Recurrent infections of the urinary and respiratory tract was the main reason for this therapy. Follow-up of 1 to 6 years showed good therapeutic effect of thymus peptides on the disease activity, alleviation of the renal manifestations and decline in recurrent infections. Recently, immunotherapy with intravenous immunoglobulins has been applied to 13 patients with IgG deficiency, renal failure, and persistent nephrotic syndrome. Preliminary results of the immunoglobulin treatment encourage to the further study on this therapeutic method. PMID- 9355681 TI - Different modes of action of high-dose immunoglobulins in rheumatoid arthritis. AB - Several clinical trials have demonstrated that in patients with rheumatoid arthritis, high doses intravenous immunoglobulins (i.v.i.g.) are frequently leading to an improvement of the morning stiffness and the extra articular symptoms, as well as to a steroid-sparing effect. This i.v.i.g. therapy seems furthermore to have immunomodulatory activities in rheumatoid arthritis, since the clinical benefit is concomitant with a reduction in the number of CD4+ T lymphocytes, with a significant dtop in the level of circulating immune complexes (CIC), and with an inhibition of the early phases of the B-lymphocytes activation. Due to the heterogeneic aspects of the rheumatoid arthritis pathogeny, it is to be expected that i.v.i.g. may interfere in different ways with many of the sequential processes in operation in the development of this disease. 1. IgGs may facilitate the reticulo endothelial clearance of inflammatory CICs by increasing their size as a consequence of the addition of exogeneous antibodies (Ab) which offers more IgG-Fc fragments as ligands for the monocytes/macrophages Fc-gamma III receptors. 2. i.v.i.g are known to down regulate the activation of B-lymphocytes and their differentiation in specialized Abs-secreting plasma-cells, and this process could affect the clones of B-cells producing anti-collagen II auto-antibodies and Abs with rheumatoid factor activity. 3. Some experimental data allow to suspect the responsibility of a super-Ag in certain forms of rheumatoid arthritis. I.v.i.g. preparations contain specific Abs which bind some of these super Ags, and impair their adequate presentation to T helper-cells. 4. Cytokines and specially the interleukin I (IL 1) which stimulates the collagenase activity, are pivotal elements in the perpetuation of the inflammatory connective tissue damages in rheumatoid arthritis. I.v.i.g. preparations contain soluble cytokine-receptors, as well as anti-IL-1 and IL-6 specific Abs which both act as cytokines agonists and sustain the effective antiinflammatory action of high-dose polyclonal IgGs. Although more clinical data are still needed to sustain the routine use of i.v.i.g. in rheumatoid arthritis, the efficacy of this therapy is not disputable any more in other pathologies with similar immunological disorders. PMID- 9355682 TI - Outlooks of systemic enzyme therapy in rheumatoid arthritis and other immunopathological diseases. AB - Systemic enzyme therapy (SET) represents a specific therapeutic approach consisting in peroral application of blends of animal and plant hydrolytic enzymes. A significant part of the swallowed enzymes (about 25%) is resorbed in the intestine in functionally active form. After being complexed with natural antiproteases, enzymes set concentrated in wounds, inflammation sites and immunopathological foci. SET has many important indications in traumatologic, thrombotic, infectious, inflammatory, immunopathologic and even tumorous processes. Rheumatoid arthritis, Bechterew's disease, activated arthrosis and extraarticular rheumatism represent important and sensitive targets of SET. In situ and in vivo studies continue to elucidate selective interferences of absorbed proteolytic enzymes with the crucial pathogenic mechanisms of rheumatoid processes. PMID- 9355684 TI - Combination therapy of juvenile rheumatoid arthritis with hydroxychloroquine-gold methotrexate: a pilot study. PMID- 9355683 TI - Cyclosporin in pediatric rheumatology; a seven years experience. PMID- 9355685 TI - Arthroscopic synovectomy of the knee joint in the treatment of patients with juvenile chronic arthritis. PMID- 9355686 TI - Therapeutical and immunological effects of methylprednisolone pulse therapy in comparison with intravenous immunoglobulin. Treatment in patients with juvenile chronic arthritis. AB - Twenty patients with polyarticular or systemic subtypes of juvenile chronic arthritis were primary treated with either Methylprednisolone (MP) pulses (group I) or i.v. Immunoglobulin (IG) (group II) in combination with Methotrexat and low dosages of Glucocorticosteroids. Clinical effects of treatment were rapid and excellent in both groups and also the regression of inflammatory activity. MP pulses and also IG-treatment decrease significant the respective part of lymphocytes and T-cells. Significant is also the decrease of CD4 and CD8 cells and the normalization of the CD4/CD8 ratio in both groups. Different are the effects on B-cells and NK-cells between the two groups. Whereas MP-pulses decrease the number of B-cells. IG-treatment leads to high increase. The number of NK-cells increases after each single MP-pulse and decreases significant after IG-infusions. PMID- 9355687 TI - Intraarticular hip treatment with triamcinolonehexacetonide in juvenile chronic arthritis. AB - To evaluate the effect and tolerance of intraarticular Triamcinolonehexacetonide (TCH) in the course of chronic coxitis in juvenile chronic arthritis (JCA) in an open uncontrolled study. Since 1990 we treated patients < 16 years of age suffering from chronic coxitis with 1 mg/TCH/KG body weight. The patients were checked again 4-8 weeks after the treatment. Clinical and ultrasound courses were recorded with the help of ultrasound and joint scores. The evaluation took place 6, 12, 24 and 36 months after the treatment. At that time we give a report on the 12 months follow-up of 37 hip joints and the 24 months follow-up of 20 hip joints. The immediate effect of TCH influencing mobility, pain sensitivity and joint effusions of the patients is impressing. The long term effect of TCH has to be evaluated by regular check ups for at least 2 years. An individual comparison with the not treated contralateral joint would be desired if ethically justifiable. Most patients suffering from polyarticular diseases with a long course of coxitis needed more than one injection of TCH (mean reinjection time 5.8 months). Any avascular necrosis of the femoral heads or other complications were not observed. PMID- 9355688 TI - Measles surveillance. PMID- 9355689 TI - Legionnaires' disease in Yorkshire and South Wales. PMID- 9355690 TI - EMRSA-16: a new epidemic strain of Staphylococcus aureus. PMID- 9355692 TI - AIDS and HIV-1 infection in the United Kingdom: monthly report. PMID- 9355691 TI - HIV transmission in men who have sex with men. PMID- 9355693 TI - Tuberculosis in Western Europe. PMID- 9355694 TI - Diphtheria immunisation for travellers to the former USSR. PMID- 9355695 TI - Reuse of cellulose acetate membrane strips for protein and haemoglobin electrophoretic analysis. AB - The exorbidant cost of electrophoretic analysis, many a times becomes the major limitation factor. As most of the clinical laboratories import the cellulose acetate membrane which costs 5 to 6 U.S. $ that is Rs. 180/- to Rs. 210/-, the final cost goes upto Rs. 350/- to Rs. 500/-. We have observed that CAN strips can be effectively reused 6 to 7 times for subsequent electrophoretic analysis. PMID- 9355696 TI - Myocarditis and hemiplegia from scorpion bite--a case report. AB - A 16 year old boy presenting with features of myocarditis and pulmonary oedema following scorpion sting developed hemiplegia with patchy vasculitic lesions on CT scan. The possible pathogenic mechanism is discussed. PMID- 9355697 TI - Hepatic cysts in adult polycystic kidney disease. AB - Of ninety two adult polycystic kidney patients 18.5% had liver cysts. Mostly they were multiple. Liver cysts were more common in men. PMID- 9355698 TI - Haematological effects of hexachlorocyclohexane (HCH) in mice--results and possibilities. AB - Adult male Swiss albino mice when exposed to a chlorinated insecticide hexachlorocyclohexane (HCH or BHC) at 100 mg/mouse/ once oral and i.p. both for acute exposures and 500 ppm and 10 ppm in food for 100 and 400 days respectively for intermediate and chronic exposures. Haematological observations revealed HCH induced anaemia. PMID- 9355699 TI - Yoga for rehabilitation: an overview. AB - The use of yoga for rehabilitation has diverse applications. Yoga practice benefited mentally handicapped subjects by improving their mental ability, also the motor co-ordination and social skills. Physically handicapped subjects had a restoration of some degree of functional ability after practicing yoga. Visually impaired children children showed a significant decrease in their abnormal anxiety levels when they practiced yoga for three weeks, while a program of physical activity had no such effect. Socially disadvantaged adults (prisoners in a jail) and children in a remand home showed significant improvement in sleep, appetite and general well being, as well as a decrease in physiological arousal. The practice of meditation was reported to decrease the degree of substance (marijuana) abuse, by strengthening the mental resolve and decreasing the anxiety. Another important area is the application of yoga (and indeed, lifestyle change), in the rehabilitation of patients with coronary artery disease. Finally, the possible role of yoga in improving the mental state and general well being of HIV positive persons and patients with AIDS, is being explored. PMID- 9355700 TI - Diabetes mellitus and glucokinase. PMID- 9355701 TI - AIIMS develops AIDS test kit. PMID- 9355702 TI - Usefulness of dark field microscopy after differential centrifugation in the early diagnosis of leptospirosis in dog and its human contacts. AB - 1. We found leptospira in the blood of two out of three police dogs by dark field microscopic examination after high speed centrifugation. One dog had fever and the other was asymptomatic. Leptospira could not be seen in the urine of one police dog which died of jaundice. 11 out of 21 human contacts were found to be positive for leptospira after low speed centrifugation and 5 after high speed centrifugation. One child had jaundice and an another child had fever. Others had mild symptoms of headache to none. Dark field microscopy after differential centrifugation is useful in the early diagnosis of leptospirosis and thereby could prevent later complications like jaundice. PMID- 9355703 TI - Glycosylation of hemoglobin and erythrocyte membrane proteins mediated changes in osmotic fragility of erythrocytes. AB - The increased levels of glycosylated hemoglobin and glycosylated erythrocyte membrane proteins in gestational diabetes could bring about alterations in osmotic fragility in erythrocytes of gestational diabetes during their first trimester. The glycosylated hemoglobin levels in erythrocytes were found to be significantly elevated with a mean of 5.42 +/- 0.77 mg/dl when compared to 3.45 +/- 0.62 mg/ dl in non diabetic pregnant control (where as in non pregnant women the value was 3.4 +/- 0.47 (n = 25)). The mean osmotic fragility (MOF) of control cells was 60.2 + 0.6 mMol/L Nacl and MOF of gestational diabetic cells was 63.93 +/- 0.6 mMol/L Nacl. The mean osmotic fragility, operationally defined as the Nacl concentration for 50% hemolysis, was found significantly higher by 3.9 + 0.01 mMol/L Nacl in gestational diabetes, than in normal cells. The total levels of glycosylated membrane proteins were increased from 50.60 +/- 8.0 in control to 69.14 + 0.47 in gestational diabetes. Therefore it is proposed that the increased levels of glycosylated Hb as well as glycosylated membrane proteins have a role in altering the membrane permeability resulting in increased osmotic fragility of erthrocytes in gestational diabetes. PMID- 9355704 TI - Food expenditure pattern, storage practices, food fads and fallacies of seven villages of Guntur districts. AB - The present study was carried out to assess the food expenditure pattern, storage practices, Food fads, and fallacies of seven villages of Guntur districts. Expenditure on protective foods in Buddam village isn nil, due to which the preschool children are in Grade II and Grade III malnutrition. Introduction of weaning food to the children in all the villages is after 6 months. The most commonly used storage structure for paddy are gunny bag, puri and patra. PMID- 9355705 TI - Cardiac metastasis from carcinoma breast--a case report. AB - Secondary neoplasms of the heart are more common than primary tumours. Metastasis occurs most commonly from bronchogenic carcinoma followed by lymphoma and carcinoma breast. Most often cardiac metastasis go undetected as they are asymptomatic and occurs as a terminal event. A 51 year old lady who developed cardiac metastasis from carcinoma breast diagnosed during life is reported with brief review of the literature. PMID- 9355706 TI - A study of some etiological factors and morbid conditions in mentally handicapped children. AB - In the present study most of the subjects belonged to mild and moderate degree of mental handicap i.e. 30.8% and 44.9% respectively while severe degree of mental handicap was present in 22.7% subject. The factors responsible for mental handicap include prenatal factors (34.6%), perinatal factors (15.1%) and post natal factors (37.9%). In 12.4% cases etiology was not known. The associated behaviour problems were observed in 33% of the study subjects while other morbid conditions were present in 47% of the study subjects. PMID- 9355707 TI - Evaluation of status of tuberculin skin sensitivity test among medical students. AB - The status of tuberculin skin sensitivity tests was evaluated in 87 medical students. It was found that Mantoux test has less significant role to play in the vaccinated individuals and in the population at high risk of exposure. Hence this test is to be supplemented with clinical manifestations and other investigations to establish the final diagnosis of tuberculosis. PMID- 9355708 TI - Relationship of erythrocyte superoxide dismutase, serum lipid peroxides and age. AB - Serum Lipid peroxides (LP) were measured as an index of free radical mediated lipid peroxidation in sixty normal healthy individuals. Erythrocyte superoxide dismutase (SOD) activity was also measured simultaneously in the subjects. The serum LP levels were found to be increased progressively with age, while erythrocyte SOD was, in contrast, found to be decreased with age. There was excellent positive correlation between serum LP and age (r = 0.959) while the erythrocyte SOD was inversely correlated with age (r = -0.845). The serum LP and erythrocyte SOD have also shown some inverse relationship (r = -0.8395). The result indicate that the balance between antioxidant & pro-oxidant factors in free radical metabolism shifts towards increased lipid peroxidation with advancing age. PMID- 9355709 TI - Metallic ion concentration during menstrual cycle in normally menstruating women. AB - Plasma concentration of metallic ions levels during menstrual cycle of twenty normally menstruating women were observed in four phases i.e. menses, follicular, ovulatory and luteal. The concentration of magnesium, zinc, selenium and manganese was highest during menses and lowest at ovulatory phase. There was rise in ionic levels of magnesium and selenium, while fall in zinc and manganese during luteal phase. Findings demonstrate changes in metallic ions (Magnesium, zinc, selenium and manganese) level in relation to hormonal status during menstrual cycle in women. PMID- 9355710 TI - Bhiwandi patients. Neuropathologic findings and toxicologic possibilities preliminary communication. PMID- 9355711 TI - Hepatitis B kills more than AIDS. PMID- 9355712 TI - Evaluation of efficacy of standard haemodialysis and verapamil added peritoneal dialysis. AB - The study was carried out on 30 adult subjects of acute renal failure who were randomly divided into two groups of 15 patients each. Group A patients were subjected to standard haemodialysis (HD) for 4 hours using holofibre dialyser and group B patients received 36 hours of peritoneal dialysis (PD) where Verapamil was added intraperitonealy in the dose of 10 mg/ L/cycle in the clearance periods III, V, VII, IX and XI (Total dose 150 mg). The 36 hours of PD consisted of 36 cycles of one hour duration each and these were divided into 12 clearance periods (CP) of 3 cycles each. Following both the forms of dialytic treatments, there was significant improvement in the signs of uraemia with fall in the blood urea and serum creatinine levels. The peritoneal clearances, percentage fall of urea and creatinine, and protein loss were similar in the two groups (p > 0.5). However, ultrafiltration was significantly higher in the group B. No untoward effects were noticed in group B however group A patients had episodes of hypotension (5)] disequilibrium (6) and cardiac arrhythmias (1). It can be concluded that both clinically and biochemically, 36 hours of Verapamil added PD and 4 hours of hemodialysis are comparable and therefore, peritoneal dialysis may be used more frequently in acute renal failure specially in situations where trained dialysis staff is not available and patients are not haemodynamically stable. PMID- 9355713 TI - A rare variation of deep femoral vein. AB - Venous pattern particularly in the lower limbs is of great clinical importance, while ligating the veins to prevent the spread of thrombus. Normally deep femoral vein drains the area supplied by profunda femoris artery, follows the pattern of branches of this artery and ends independently into the femoral vein. However in few cases it establishes communication with popliteal vein of femoral vein at a lower level. In the present study the deep femoral vein replaced most of the popliteal vein by establishing communication with the tibial veins. PMID- 9355714 TI - Dairy income and somatic status of pre-school children. AB - The study was conducted on pre school children of Karnataka to assess the impact of additional income from dairy on somatic status of children. Analysis of data showed that there were no differences with respect to height and weight of children between the study groups. Hence, it was concluded that income from agriculture and income from dairy products either independently or jointly did not influence height for age, weight for age and weight for height for age of the child. PMID- 9355715 TI - Distribution of malarial parasites: effect of gender of construction workers. AB - The city of Mangalore in South India was having increasing number of malaria cases from 1990. Concerned over the import of cases through migrant construction workers, a screening was done among them using clinical and parasitological methods. This demonstrated 6.28% slide positivity rate with statistically insignificant difference in prevalence of infection between males and females. There were many asymptomatic individuals reporting positive only on peripheral smear examination. Yet, clinical symptoms like fever were found to have good predictive value on logistic regression. It was more so with the P. falciparum which is a relatively new entrant to Mangalore. PMID- 9355716 TI - Self medication--a growing concern. AB - 31% persons practiced self medication. They were more in 31-40 years (26.9%) and 41-50 years (30.8%). Males were more than females in self prescribers. Illiterate (23.1%) and graduates (26%) were more common self medicators, while labourers (26%) and business men (19.3%) were more involved in self medication. Fever (17.4%), cough (22.2%), boils (7.6%) and acidity (6.8%) were the common complaints for self-prescription. Thus analgesics and antipyretics, (30.9%), tonics (16.1) and antibiotics (10.7%) were commonly used drugs by the customers. Most of the information of drugs was from friends or neighbours (30.8%) or chemist (23.1%). PMID- 9355717 TI - Solitary cavernous haemangioma of the jejunum with intra-abdominal bleeding--case report. PMID- 9355718 TI - Indian clinical pathology laboratories; quality of measure! PMID- 9355719 TI - Distribution of agenesis of palmaris longus muscle in 12 to 18 years old age groups. AB - In Gaziantep, a survey was made on 7000 students whose ages changing from 12 to 18, 1000 individuals for each age group. The tendon examination method about Palmaris Longus Muscle (PLM) agenesis was used in this study. In girls, unilateral PLM agenesis was found to be 23 percent. Bilateral PLM agenesis was 45.3 percent. In Boys unilateral and bilateral PLM agenesis was found to be 19.5 and 42.1 percent respectively. The overall percentage of agenesis in the total number of individual was 63.9 percent. PMID- 9355720 TI - HIV associated chronic atypical osteomyelitis by mycobacterium fortuitum- Chelonae group--a case report. AB - In developing countries where tuberculosis is endemic, the HIV patients have tuberculosis as one of the major opportunistic infections. Commonly M. tuberculosis, M. Avium-intracellular are the causative agents of pulmonary, extra pulmonary and disseminated infection in AIDS patients. Here is a report of a 32 year old HIV positive male who presented as chronic atypical osteomyelitis of right tibia. Core biopsy confirmed the diagnosis by histopathology, direct microscopy and culture of M. fortuitum-chelonae group identified by the biochemical tests. TB IgG ELISA was strongly positive. ELISA for HIV antibodies was reactive on three occasions. Western blot was positive for HIV-1 antibodies. Patient responded well to ciproflox and antitubercular treatment and is currently under a follow up. PMID- 9355721 TI - Chronic back pain--a review. PMID- 9355723 TI - Effect of PMA and IL-1 on matrix proteins with special reference to kidney mesangial cells. AB - Cytokines have different effects on the extracellular matrix proteins which were produced by a number of different cell lines. The extracellular matrix proteins fibronectin, collagen IV, vitronectin, laminin, proteoglycan, fetal proteoglycan and actin; and the cells human mesangial, BeWo, Hep G2 and BHK using APAAP and dot-blot assay were used. It was shown that IL-1 and PMA increased the staining intensity of proteoglycan, fibronectin and collagen IV and vitronectin and did not alter the intensity of staining. While PMA and IL-1 decreased but did not alter laminin, PMA increased the staining intensity of proteoglycan in mesangial cells but not altered in mesangial cells stimulated with IL-1. These results show that PMA and IL-1 altered the production and secretion of extracellular matrix proteins, these in turn could lead to the alteration of pore size, matrix properties and filtration of mesangial cells, and thus influence pathological conditions of renal diseases. PMID- 9355722 TI - Regression analysis of the effect of full and partial lactation on post-partum amenorrhoea in lower middle class Indian females. AB - This study presents a simple regression analysis of the impact of full and partial breast feeding on post-partum amenorrhoea (PPA) in lower middle class Indian women. The study has been done on a sample of hospital based data collected in 1992. The analysis shows a highly significant correlation. Further, the impact of lactation on PPA was notably less in case with partial breast feeding as compared to those with full breast feeding. PMID- 9355724 TI - Tuberculous duodenal obstruction--a case report. AB - Proximal duodenal obstruction due to tuberculosis can masquarade as duodenal ulcer. Although commonest cause of duodenal obstruction is ulcer, other causes must be considered, particularly tuberculosis which is common in tropics. PMID- 9355725 TI - Infectious diarrhoea--an update. AB - Infectious diarrhoea is one of the most common illness affecting mankind. Recent advancement have led to detailed understanding of causative agents and the pathogenesis of the infections. Fluid and electrolyte replacement remains the most important aspect of treatment. The role of antimicrobial agents is discussed. PMID- 9355726 TI - The good news about AIDS. PMID- 9355728 TI - A novel procedure for the efficient purification of the cystic fibrosis transmembrane conductance regulator (CFTR). AB - This report describes a novel, single-step strategy for the purification of the cystic fibrosis transmembrane conductance regulator from Sf9 cells, which will facilitate studies of the structure-function relationships of this clinically important molecule. The new method combines the use of the novel detergent sodium pentadecafluoro-octanoate with metal-affinity chromatography to produce a high yield of purified protein which can be functionally reconstituted as a chloride channel and an ATPase. PMID- 9355729 TI - Novel activity of endothelin-converting enzyme: hydrolysis of bradykinin. AB - Endothelin-converting enzyme (ECE) is the key enzyme in the production of the potent vasoconstrictor endothelin from its inactive precursor big endothelin. To date, no other physiological peptide substrate has been identified for ECE. Here, by using Chinese hamster ovary (CHO) cells transfected with rat ECE-1 cDNA, we have established that ECE can hydrolyse the vasodilator bradykinin. The hydrolysis of bradykinin by ECE is exclusively at the Pro7-Phe8 bond, producing bradykinin-(1-7) and bradykinin-(8-9). Hydrolysis is completely inhibited by 100 microM phosphoramidon and 200 microM EDTA, but only slightly by the specific neprilysin inhibitor thiorphan (100 microM). The ability of ECE to act as a peptidyl dipeptidase rather than an endopeptidase in hydrolysing bradykinin suggests a much broader specificity for the enzyme than previously recognized, which may lead to the design of new and specific inhibitors of ECE and to the identification of other potential physiological substrates. PMID- 9355727 TI - Syndecans: multifunctional cell-surface co-receptors. AB - This review will summarize our current state of knowledge of the structure, biochemical properties and functions of syndecans, a family of transmembrane heparan sulphate proteoglycans. Syndecans bind a variety of extracellular ligands via their covalently attached heparan sulphate chains. Syndecans have been proposed to play a role in a variety of cellular functions, including cell proliferation and cell-matrix and cell-cell adhesion. Syndecan expression is highly regulated and is cell-type- and developmental-stage-specific. The main function of syndecans appears to be to modulate the ligand-dependent activation of primary signalling receptors at the cell surface. Principal functions of the syndecan core proteins are to target the heparan sulphate chains to the appropriate plasma-membrane compartment and to interact with components of the actin-based cytoskeleton. Several functions of the syndecans, including syndecan oligomerization and actin cytoskeleton association, have been localized to specific structural domains of syndecan core proteins. PMID- 9355730 TI - Hydrolysis of somatostatin by human tissue kallikrein after the amino acid pair phe-Phe. AB - Somatostatin-(1-14) was hydrolysed by human tissue kallikrein at the Phe7-Trp8 bond, after a Phe-Phe pair of amino acids, with similar kinetic parameters to those described for human high- and low-molecular-mass kininogens. Substance P and human insulin, which also contain a Phe-Phe pair in their sequences, were both resistant. More details of this hydrolytic specificity of human tissue kallikrein were obtained by synthesizing and assaying internally quenched fluorescent peptides containing the sequence of somatostatin-(1-14), as well as the reactive-centre loop of human kallikrein-binding protein (kallistatin). We also observed that human tissue kallikrein hydrolysed growth hormone-releasing hormone at the Arg11-Lys12 bond, although this peptide contains in its structure a pair of leucines (Leu22-Leu23), in contrast with the Phe-Phe pair in somatostatin. We have also demonstrated the susceptibility to human tissue kallikrein of some chromogenic peptide s with the general structure of X-Phe-Phe p-nitroanilide and D-Pro-Phe-Phe-4-methylcoumaryl-7-amide. PMID- 9355731 TI - Reduction of equilibrative nitrobenzylthioinosine-sensitive nucleoside transporter in tamoxifen-treated MCF-7 cells: an oestrogen-reversible phenomenon. AB - MCF-7 cells display both nitrobenzylthioinosine (NBMPR)-sensitive (es) and NBMPR insensitive (ei) equilibrative, but not the Na+-dependent, nucleoside transport. Transport of uridine by es is more sensitive to inhibition by purine nucleosides, whereas the ei component is more sensitive to nucleosides without an amino side group, such as inosine and thymidine. When exposed to 10 microM tamoxifen for 5 days, MCF-7 cells displayed a 44% decrease in the total number of NBMPR-binding sites [Bmax from 245000+/-18000 to 136000+/-25000 sites per cell (mean+/-S.E.M.; n=5; P<0.05)], and a 57% decrease in cell growth with no significant change in binding affinities [Kd from 0.37+/-0.05 to 0.45+/-0.08 nM (n=5; P>0.05)]. Kinetic studies of [3H]uridine transport showed a decrease in the Vmax of the es component from 21.7+/-0.3 (n=8) to 8.4 +/- 2.2 microM/s (n=4; P < 0.05), whereas the Vmax of the ei component [from 4.7 +/- 0.5 (n=8) to 5.8 +/- 1.6 microM/s (n=4; P > 0.05)] and Km values for both components [es from 460 +/- 80 to 630 +/- 280 microM (n>/=4; P > 0.05) and ei from 355 +/- 115 to 440 +/- 220 microM (n>/=4; P > 0.05)] did not change significantly. Oestradiol at 100 nM reversed almost completely the NBMPR-binding site decrease and growth retardation in tamoxifen-treated cells. Thus tamoxifen is shown to cause an oestrogen-reversible decrease of es nucleoside transporters in MCF-7 cells. PMID- 9355733 TI - Lysosomal alpha-mannosidases of mouse tissues: characteristics of the isoenzymes, and cloning and expression of a full-length cDNA. AB - Lysosomal alpha-d-mannosidase from mouse tissues was separated into its constituent isoenzymes by DEAE-cellulose chromatography. Forms corresponding to the human isoenzymes B and A were present in testis, brain, spleen and kidney, whereas in epididymis and liver only the B form was present. Murine alpha mannosidases A and B are glycoproteins and have pH optima, thermal stabilities and molecular masses similar to those of the human isoenzymes. A full-length cDNA (3.1 kb) containing the complete coding sequence for alpha-mannosidase was isolated from a mouse macrophage cDNA library. Comparison of the deduced amino acid sequences of human and mouse alpha-mannosidases showed that they had 75% identity and 83% similarity. Expression of this cDNA in COS cells showed that both the A and the B isoenzymes can arise from a single transcript. Northern blotting analysis showed a 10-fold range in the abundance of alpha-mannosidase mRNA in mouse tissues, with the highest levels found in epididymis, and the lowest in liver. PMID- 9355732 TI - Angiotensin-converting enzyme secretase is inhibited by zinc metalloprotease inhibitors and requires its substrate to be inserted in a lipid bilayer. AB - Mammalian angiotensin-converting enzyme (ACE; EC 3.4.15.1) is one of several proteins that exist in both membrane-bound and soluble forms as a result of a post-translational proteolytic processing event. For ACE we have previously identified a metalloprotease (secretase) responsible for this proteolytic cleavage. The effect of a range of structurally related zinc metalloprotease inhibitors on the activity of the secretase has been examined. Batimastat (BB94) was the most potent inhibitor of the secretase in pig kidney microvillar membranes, displaying an IC50 of 0.47 microM, whereas TAPI-2 was slightly less potent (IC50 18 microM). Removal of the thienothiomethyl substituent adjacent to the hydroxamic acid moiety or the substitution of the P2' substituent decreased the inhibitory potency of batimastat towards the secretase. Several other non hydroxamate-based collagenase inhibitors were without inhibitory effect on the secretase, indicating that ACE secretase is a novel zinc metalloprotease that is realted to, but distinct from, the matrix metalloproteases. The full-length amphipathic form of ACE was labelled selectively with 3-trifluoromethyl-3-(m [125I]iodophenyl)diazirine in the membrane-spanning hydrophobic region. Although trypsin was able to cleave the hydrophobic anchoring domain from the bulk of the protein, there was no cleavage of full-length ACE by a Triton X-100-solubilized pig kidney secretase preparation when the substrate was in detergent solution. In contrast, the Triton X-100-solubilized secretase preparation released ACE from pig intestinal microvillar membranes, which lack endogenous secretase activity, and cleaved the purified amphipathic form of ACE when it was incorporated into artificial lipid vesicles. Thus the secretase has an absolute requirement for its substrate to be inserted in a lipid bilayer, a factor that might have implications for the development of cell-free assays for other membrane protein secretases. ACE secretase could be solubilized from the membrane with Triton-X 100 and CHAPS, but not with n-octyl beta-D-glucopyranoside. Furthermore trypsin could release the secretase from the membrane, implying that like its substrate, ACE, it too is a stalked integral membrane protein. PMID- 9355734 TI - Carbon dioxide and light regulation of promoters controlling the expression of mitochondrial carbonic anhydrase in Chlamydomonas reinhardtii. AB - Nuclear genes coding for carbonic anhydrase, a major mitochondrial constituent in Chlamydomonas reinhardtii grown under limited CO2, were characterized. Two genes, ca1 and ca2, were found within 7 kb of genomic DNA, organized 'head to head' in a large inverted repeat. The DNA sequences for the two genes were very similar, even in the promoter regions and in introns, indicating that the repeat is a result of a recent duplication. To study gene regulation, elements from the upstream region of ca1 were fused to the arylsulphatase reporter gene. After transformation, the expression of arylsulphatase was regulated similarly to the endogenous ca1/ca2 genes, even when the promoter was trimmed down to 194 nt. Expression could not be detected when 5% CO2 was bubbled into the growth medium, but was induced within hours after transfer to air. The ca1 promoter was not induced in low light, but at intermediate light levels its activity was dependent on the irradiance. O2 concentration had no effect on the promoter activity, indicating that photorespiratory metabolites are not triggering the response. The availability of cells transformed with a CO2-regulated reporter gene should facilitate further studies on the metabolic adaptations that occur in some green algae in response to the external CO2 level. PMID- 9355735 TI - Delta-1-piperideine-6-carboxylate dehydrogenase, a new enzyme that forms alpha aminoadipate in Streptomyces clavuligerus and other cephamycin C-producing actinomycetes. AB - Delta-1-Piperideine-6-carboxylate (P6C) dehydrogenase activity, which catalyses the conversion of P6C into alpha-aminoadipic acid, has been studied in the cephamycin C producer Streptomyces clavuligerus by both spectrophotometric and radiometric assays. The enzyme has been purified 124-fold to electrophoretic homogeneity with a 26% yield. The native protein is a monomer of 56.2 kDa that efficiently uses P6C (apparent Km 14 microM) and NAD+ (apparent Km 115 microM), but not NADP+ or other electron acceptors, as substrates. The enzyme activity was inhibited (by 66%) by its end product NADH at 0.1 mM concentration. It did not show activity towards pyrroline-5-carboxylate and was separated by Blue-Sepharose chromatography from pyrroline-5-carboxylate dehydrogenase, an enzyme involved in the catabolism of proline. P6C dehydrogenase reached maximal activity later than other early enzymes of the cephamycin pathway. The P6C dehydrogenase activity was decreased in ammonium (40 mM)-supplemented cultures, as was that of lysine 6 amino-transferase. P6C dehydrogenase activity was also found in other cephamycin C producers (Streptomyces cattleya and Nocardia lactamdurans) but no in actinomycetes that do no produce beta-lactams, suggesting that it is an enzyme specific for cephamycin biosynthesis, involved in the second stage of the two step conversion of lysine to alpha-aminoadipic acid. PMID- 9355736 TI - Specificity of the hyaluronate lyase of group-B streptococcus toward unsulphated regions of chondroitin sulphate. AB - The purification and properties of a hyaluronate lyase secreted by Streptococcus agalactiae, which is believed to facilitate the invasion of host tissues by the organism, have been described previously [Pritchard, Lin, Willingham and Baker (1994) Arch. Biochem. Biophys. 315, 431-436]. The specificity of the limited cleavage of chondroitin sulphate by the enzyme is the subject of this report. To simplify the task, a chondroitin sulphate from the Swarm rat chondrosarcoma, which contains only 4-sulphated and unsulphated disaccharide repeats, was used in this study. Tetrasaccharides from an ovine testicular hyaluronidase digest of the chondroitin sulphate were isolated, identified and tested as substrates of the streptococcal hyaluronate lyase. Only tetrasaccharides with an unsulphated disaccharide at the reducing end were cleaved (by elimination at the N acetylgalactosaminidic bond). Thus chondroitin sulphate chains are cleaved by the action of this lyase at every unsulphated disaccharide repeat, but release of unsaturated unsulphated disaccharides only occurs from sites where two or more sequential unsulphated disaccharide repeats are present. Analysis of the chondrosarcoma chondroitin sulphate showed that of approximately five unsulphated disaccharide repeats per chain, two are clustered. The ability of group-B streptococcal hyaluronate lyase to cleave chondroitin sulphate may allow the organisms to invade tissues more efficiently. The demonstrated specific and highly limited cleavage of chondroitin sulphate by this bacterial lyase promises to be a useful tool in the determination of chondroitin sulphate structure and variability. PMID- 9355738 TI - Sequence of human carboxypeptidase D reveals it to be a member of the regulatory carboxypeptidase family with three tandem active site domains. AB - We have cloned the cDNA for human carboxypeptidase D (CPD), a new B-type metallocarboxypeptidase that is membrane bound and has an acidic pH optimum. The 5.8 kb of cDNA sequenced contains an open reading frame of 4131 bp encoding 1377 amino acid residues. The sequence is similar (75% identity) to duck gp180, a protein that was isolated, cloned and sequenced as a hepatitis B virus-binding protein but not characterized as a carboxypeptidase. Hydropathic analysis revealed a hydrophobic region at the N-terminus, representing the signal peptide, and one near the C-terminus that probably represents the transmembrane anchor. The most striking feature is the presence of three tandem carboxypeptidase homology domains that have sequence similarity to the regulatory B-type carboxypeptidase family, typified by carboxypeptidases M, E and N. Because of the three repeats, CPD is about three times larger (175-180 kDa) than other members of this family (approx. 50-62 kDa). Domain 2 is most closely related to carboxypeptidases M, E and N (45-48% identity), followed by domain 1 (37-38%) and domain 3 (20-27%). There is a much higher sequence identity in regions containing putative active site residues, and all catalytically important residues are strictly conserved in domains 1 and 2. In domain 3, however, only 1 of 8 active site residues is conserved, indicating that this portion might not be catalytically active. Northern blotting of mRNA from human tissues and cells showed high levels of CPD mRNA in placenta, pancreas and Hep G2 hepatoma cells, and smaller amounts in skeletal muscle, heart and HT-29 colon carcinoma and melanoma cell lines. PMID- 9355737 TI - Stat1 associates with c-kit and is activated in response to stem cell factor. AB - Interaction of stem cell factor (SCF), a haematopoietic growth factor, with the receptor tyrosine kinase c-kit leads to autophosphorylation of c-kit as well as tyrosine phosphorylation of various substrates. Little is known about the role of the JAK/STAT pathway in signal transduction via receptor tyrosine kinases, although this pathway has been well characterized in cytokine receptor signal transduction. We recently found that the Janus kinase Jak2 associates with c-kit and that SCF induces rapid and transient phosphorylation of Jak2. Here we present evidence that SCF activates the transcription factor Stat1. Phosphorylated c-kit co-immunoprecipitates with Stat1 within 1 min of SCF stimulation of the human cell line MO7e. Co-precipitation experiments using glutathione S-transferase fusion proteins indicate that association with c-kit is mediated by the Stat1 SH2 domain. Stat1 is rapidly tyrosine-phosphorylated in response to SCF in MO7e cells, the murine cell line FDCP-1 and normal progenitor cells. SCF-induced phosphorylation of Jak2 and Stat1 was also observed in murine 3T3 fibroblasts stably transfected with full-length human c-kit receptor. Furthermore c-kit directly phosphorylates Stat1 fusion proteins in in vitro kinase assays. Electrophoretic mobility-shift assays with nuclear extracts from SCF-stimulated cell lines and normal progenitor cells indicate that activated Stat1 binds the m67 oligonucleotide, a high-affinity SIE promoter sequence. These results demonstrate that Stat1 is activated in response to SCF, and suggest that Stat1 is a component of the SCF signal-transduction pathway. PMID- 9355739 TI - Expression, purification and characterization of recombinant caprine N acetylglucosamine-6-sulphatase. AB - Mucopolysaccharidosis type IIID or Sanfilippo D syndrome is a lysosomal storage disorder caused by the deficiency of N-acetylglucosamine-6-sulphatase (Glc6S). In addition to human patients, a Nubian goat with this disorder has been described and the caprine Glc6S (cGlc6S) cDNA cloned. In this study, the full-length cGlc6S cDNA was inserted into the expression vector, pEFNeo, which placed the cGlc6S cDNA under the transcriptional control of the human polypeptide chain elongation factor promoter. The pEFNeo expression vector also contains the human growth hormone polyadenylation signal and the genes encoding resistance to ampicillin and G418. The cGlc6S expression construct was electroporated into Chinese hamster ovary (CHO-K1) cells, and stably transfected clones were isolated. One clone, CHOrcGlc6S.17, which secreted the highest Glc6S activity into the culture medium, was selected and cultured in cell factories. The secreted recombinant cGlc6S (rcGlc6S) precursor was purified to homogeneity from conditioned medium by a two column procedure which consisted of a Cu2+-chelating Sepharose column followed by TSK G3000SW gel filtration. The native molecular mass of rcFlc6S was estimated to be 102 kDa and the subunit size was 94 kDa. The kinetic properties of cGlc6S were similar to those of human Glc6S isolated from liver. rcGlc6S was endocytosed by fibroblasts from patients with mucopolysaccharidosis type IIID via the mannose 6 phosphate receptor-mediated pathway resulting in correction of the storage phenotype of these cells. PMID- 9355741 TI - Molecular cloning, chromosomal assignment and tissue-specific expression of a murine alpha(1,2)fucosyltransferase expressed in thymic and epididymal epithelial cells. AB - Terminal Fucalpha(1-2)Galbeta epitopes have been proposed to play significant roles in cell-cell interactions in development, cell adhesion, and malignant transformation. To begin to investigate the regulation and function of alpha(1 2)fucosylated epitopes in an animal model, we have isolated and characterized a mouse genomic DNA segment encoding a protein orthologous to the human H blood group locus alpha(1,2)fucosyltransferase (FUT1). This segment maintains an open reading frame encoding 376 amino acids sharing 75% sequence identity with the enzyme encoded by human FUT1, and 55% sequence identity with the enzyme encoded by the human Secretor blood group locus (FUT2). Expression of the open reading frame in COS-7 cells yields an alpha(1,2)fucosyltransferase activity with a Km of 7.6 mM for phenyl-beta-d-galactoside. Southern blotting and interspecific backcross analyses indicate that this murine locus represents a single copy sequence mapping to a novel locus 2.1 centimorgans from the Klk1 locus, in a region of homology between mouse chromosome 7 and the human FUT1 locus on the long arm of chromosome 19. Mouse FUT1 yields a 2.8 kb mRNA transcript identifiable in many organs, including thymus, lung, stomach, pancreas, small intestine, colon, uterus and epidiymis. Hybridization analyses in situ localize expression of FUT1 transcripts to thymic medullary and epididymal epithelial cells, implying that this gene determines the expression of cell surface Fucalpha(1-2)Galbeta epitopes in these tissues. PMID- 9355740 TI - Functional diversity and interactions between the repeat domains of rat intestinal lactase. AB - Lactase-phlorizin hydrolase (LPH), a major digestive enzyme in the small intestine of newborns, is synthesized as a high-molecular-mass precursor comprising four tandemly repeated domains. Proteolytic cleavage of the precursor liberates the pro segment (LPHalpha) corresponding to domains I and II and devoid of known enzymic function. The mature enzyme (LPHbeta) comprises domains III and IV and is anchored in the brush border membrane via a C-terminal hydrophobic segment. To analyse the roles of the different domains of LPHalpha and LPHbeta, and the interactions between them, we have engineered a series of modified derivatives of the rat LPH precursor. These were expressed in cultured cells under the control of a cytomegalovirus promoter. The results show that recombinant LPHbeta harbouring both domains III and IV produces lactase activity. Neither domain III nor IV is alone sufficient to generate active enzyme, although the corresponding proteins are transport-competent. Tandem duplication of domains III or IV did not restore lactase activity, demonstrating the separate roles of both domains within LPHbeta. Further, the development of lactase activity did not require LPHalpha; however, LPHalpha potentiated the production of active LPHbeta but the individual LPHalpha subdomains I and II were unable to do so. Lactase activity and targeting required the C-terminal transmembrane anchor of LPH; this requirement was terminal transmembrane anchor or LPH; this requirement was not satisfied by the signal/anchor region of another digestive enzyme: sucrase isomaltase. On the basis of this study we suggest that multiple levels of intramolecular interactions occur within the LPH precursor to produce the mature enzyme, and that the repeat domains of the precursor have distinct and specific functions in protein processing, substrate recognition and catalysis. We propose a functional model of LPHbeta in which substrate is channelled from an entry point located within domain II to the active site located in domain IV. PMID- 9355742 TI - Homodimerization and hetero-oligomerization of the single-domain trefoil protein pNR-2/pS2 through cysteine 58. AB - The single-domain human trefoil proteins [pNR-2/pS2 and human intestinal trefoil factor (hITF)] have seven cysteine residues, of which six are involved in maintaining the structure of the trefoil domain. The seventh does not form part of the trefoil domain and is located three residues from the C-terminus. The ability of the pNR-2/pS2 single trefoil domain protein to dimerize was examined by using recombinant protein with either a cysteine or a serine residue at this position by equilibrium ultracentrifugation, laser-assisted desorption MS, gel filtration and PAGE. pNR-2/pS2 Cys58 formed dimers, whereas pNR-2/pS2 Ser58 did not. Experiments in which the dimer was treated with thiol agents demonstrated that the dimer was linked via a disulphide bond and that the intermolecular disulphide bond was more susceptible to reduction than the intramolecular disulphide bonds. To examine whether dimeric pNR-2/pS2 was secreted by oestrogen responsive breast cancer cells, which are known to express pNR-2/pS2 mRNA, conditioned medium was separated on non-denaturing polyacrylamide gels, transferred to PVDF membrane and reacted with antiserum against pNR-2/pS2. Monomeric and dimeric pNR-2/pS2 were detected but the majority of the protein reactivity was associated with a larger protein. Treatment of this protein with thiol agents suggested that it is an oligomer containing pNR-2/pS2 linked to another protein by a disulphide bond. These studies suggest that the biological action of pNR-2/pS2 single-domain trefoil protein might involve the formation of homodimers or oligomers with other proteins. PMID- 9355743 TI - Conversion of dihydroceramide into ceramide: involvement of a desaturase. AB - Ceramide has been suggested to be a potent bioactive lipid involved in cell growth, differentiation and apoptosis. Its precursor, dihydroceramide, does not affect these processes. The truncated dihydroceramide analogues N-hexanoyl-[4,5 3H]-d-erythro-sphinganine and N-[1-14C]-hexanoyl-d-erythro-sphinganine were used to study the conversion of dihydroceramide into ceramide by rat hepatocytes. The formation of tritiated water after the addition of the tritiated substrate to intact and permeabilized rat hepatocytes was followed to measure enzyme activity. Desaturation was severely depressed in permeabilized hepatocytes, suggesting loss of cofactors. Of a variety of cofactors tested in the permeabilized cells, NADPH appeared to be stimulatory, pointing to the involvement of a desaturase. In agreement with this, the addition of inhibitors and redox effectors known to affect Delta9-stearoyl-CoA desaturase and Delta1-plasmanyl-ethanolamine desaturase to intact cells resulted in severe inhibition of the desaturation. When added to permeabilized cells fortified with NADPH, these compounds counteracted the NADPH stimulation. The enzyme system was further studied in broken cells. On cell fractionation, the activity was recovered in the microsomal fraction. The results indicate that the conversion of dihydroceramide into ceramide is ctalysed by a desaturase and not by a dehydrogenase or an oxidase as was generally believed. PMID- 9355744 TI - Different modes of interaction of pulmonary surfactant protein SP-B in phosphatidylcholine bilayers. AB - Pulmonary surfactant-associated protein B (SP-B) has been incorporated into vesicles of dipalmitoyl phosphatidylcholine (DPPC) or egg yolk phosphatidylcholine (PC) by two different procedures to characterize the dependence of lipid-protein interactions on the method of reconstitution. In method A the protein was dissolved in a small volume of either methanol or 60% (v/v) acetonitrile and injected into an aqueous phase containing phospholipid vesicles. In method B the vesicles were prepared by injection of a mixture of phospholipid and SP-B dissolved in methanol or aqueous acetonitrile. Both methods of reconstitution led to the extensive interaction of SP-B with PC bilayers as demonstrated by co-migration during centrifugation, marked protection against proteolysis, change in the fluorescence emission intensity of SP-B, and protection of SP-B tryptophan fluorescence from quenching by acrylamide. SP-B promoted the rapid adsorption of DPPC on an air/liquid interface irrespective of the method of protein reconstitution. However, the interfacial adsorption activity of SP-B reconstituted by method B remained stable for hours, but that of SP-B prepared by method A decreased with time. Electron microscopy showed that the injection of SP-B into an aqueous phase containing PC or DPPC vesicles (method A) induced a rapid aggregation of vesicles. By contrast, a much longer time was required for detecting vesicle aggregation when the protein was reconstituted by co-injection of SP-B and phospholipids (method B). The presence of 5% (w/w) SP-B in DPPC bilayers prepared by method B broadened the differential scanning calorimetry thermogram and decreased the enthalpy of the transition. In contrast, the injection of SP-B into preformed DPPC vesicles (method A) did not influence the gel-to-liquid phase transition of DPPC bilayers. Taken together, these results indicate that the mode and extent of interaction of SP-B with surfactant phospholipids depends on the conditions of preparation of lipid/protein samples, and that care should be taken in the interpretation of findings from reconstituted systems on the role of these surfactant proteins in the alveolar space. PMID- 9355745 TI - Protein tyrosine phosphatase 1B interacts with and is tyrosine phosphorylated by the epidermal growth factor receptor. AB - We used a substrate-trapping technique to search for substrates of protein tyrosine phosphatase (PTP) 1B. A catalytically inactive form of this enzyme forms a stable, phosphotyrosine-dependent complex with epidermal growth factor receptor (EGFR) both in vitro and in cells. PTP1B also interacts with activated platelet derived growth factor receptor (PDGFR) but not with colony-stimulating factor 1 receptor (CSF-1R). After binding to EGFR, PTP1B becomes tyrosine-phosphorylated at Tyr-66, a site that conforms to the consensus binding sequence for the Src homology 2 (SH2) domains of the adapter protein Grb2. This tyrosine phosphorylation is correlated with a 3-fold increase in PTP catalytic activity. These findings suggest that PTP1B selectively regulates specific activated receptor protein tyrosine kinases (RPTKs) in vivo and might itself be regulated by such receptors. PMID- 9355746 TI - Novel bradykinin signalling events in PC-12 cells: stimulation of the cAMP pathway leads to cAMP-mediated translocation of protein kinase Cepsilon. AB - In the rat pheochromocytoma cell line PC-12, bradykinin (BK) stimulated phosphatidylinositol hydrolysis by 4-5-fold and, additionally, intracellular cAMP accumulation by approx. 1.6-fold. EC50 values for BK were 3 nM and 2 nM respectively. The BK-induced increase in cAMP accumulation was paralleled by a 1.6-fold increase in protein kinase A (PKA) activity. The time course of BK stimulated inositol phosphate formation was rapid (t1/2<1 min), whereas the BK induced cAMP accumulation was lagging (t1/2 approx. 6 min). The effect of BK on the cAMP pathway was independent of pertussis toxin, excluding an indirect stimulation of adenylate cyclase via betagamma-complexes from Gi or Go proteins. Two different protein kinase C (PKC) inhibitors, bisindolylmaleimide and Ro 31 820, failed to prevent BK-induced cAMP accumulation, and exclude PKC as mediator of BK action on adenylate cyclase. In contrast, the stimulatory effect of BK on cAMP accumulation was completely abolished by two calmodulin antagonists, chlorpromazine and ophiobolin, suggesting an indirect, Ca2+/calmodulin-mediated effect of BK on the cAMP pathway. In addition, exposure of PC-12 cells to BK resulted in a translocation of the PKC isoforms alpha, delta, epsilon and zeta displaying different kinetics. The BK-induced translocations of the PCDs alpha and delta were rapid and biphasic, whereas the PKCs epsilon and zeta revealed a slower and slightly transient translocation in response to BK. The BK-elicited translocation of PKCepsilon, but not that of the PKCs alpha, delta and zeta, was prevented by two different inhibitors of adenylate cyclase, 2',5' dideoxyadenosine and MDL-12,330A, as well as the PKA inhibitor adenosine 3':5' monophosphothioate. These findings suggest that the BK-induced translocation of novel (n)PKCepsilon is mediated via the cAMP pathway. Since nPKCepsilon appears to regulate neurite outgrowth in PC-12 cells [Hundke, McMahon, Dadgar and Messing (1995) J. Biol. Chem. 270, 30134-30140] our results provide evidence for a novel signalling mechanism that might be involved in BK-induced neuronal differentiation of PC-12 cels. PMID- 9355748 TI - Expression, purification and characterization of Arabidopsis thaliana acetohydroxyacid synthase. AB - Acetohydroxyacid synthase (EC 4.1.3.18) is the enzyme that catalyses the first step in the synthesis of the branched-chain amino acids valine, leucine and isoleucine. The AHAS gene from Arabidopsis thaliana with part of the chloroplast transit sequence removed was cloned into the bacterial expression vector pT7-7 and expressed in the Escherichia coli strain BL21(DE3). The expressed enzyme was purified by an extensive procedure involving (NH4)2SO4 fractionation followed by hydrophobic and anion-exchange chromatography. The purified enzyme appears as a single band on SDS/PAGE with a molecular mass of about 61 kDa. On gel filtration the enzyme is a dimer, migrating as a single peak with molecular masses of 109 and 113 kDa in the absence and presence of FAD respectively. Ion spray MS analysis yielded a mass of 63864 Da. The enzyme has optimum activity in the pH range 6.5-8.5 and exhibits absolute dependence on the three cofactors FAD, Mg2+ and thiamine diphosphate for activity. It displays negatively co-operative kinetics with respect to pyruvate concentration. A model was derived to explain the non-hyperbolic substrate-saturation curve, involving interaction between the active sites of the dimer. The Km for the first active site was found to be 8.01 +/- 0.66 mM; the Km for the second active site could not be accurately determined but was estimated to be approx. 100 mM. The enzyme is insensitive to valine, leucine and isoleucine but is strongly inhibited by the sulphonylurea herbicide, chlorsulphuron, and the imidazolinone herbicide, imazapyr. Inhibition by both herbicides exhibits slow-binding kinetics, whereas chlorsulphuron also shows tight-binding inhibition. PMID- 9355749 TI - Role of Mg2+ in the structure and activity of maize (Zea mays L.) isocitrate lyase: indications for hysteretic behaviour. AB - The role of Mg2+ in the structure and activity of maize isocitrate lyase has been studied by CD, limited proteolysis, protection by ligands against inactivation, and activity measurements at various metal concentrations. From CD and trypsinolysis experiments, the existence of high-affinity binding sites for Mg2+ was demonstrated, and a KdME of 200 microM was determined. Both free enzyme (E) and enzyme molecules with high-affinity sites occupied (ME) are catalytically competent, the former showing 40% of the activity of the latter. Mg2+ thus acts as a non-essential activator. A second Mg2+-binding site with a KdMEM of 6 mM was revealed from protection experiments by increasing Mg2+ concentrations against inactivation. From activity measurements at different Mg2+ concentrations, the affinity of the enzyme for the Mg2+-isocitrate complex (MI) was determined to be KdE(MI) = 9 microM. Maize isocitrate lyase was shown to display hysteretic behaviour. Filling the low-affinity binding sites with Mg2+ induces a conformational change in the high-affinity binding sites resulting in an even higher affinity for Mg2+ (KdME* = 40 microM). On lowering the Mg2+ concentration again, the enzyme only responds slowly: the time needed for all enzyme molecules to return to the conformation at which KdME is 200 microM was found to be 60 min. Finally it was shown that the high-affinity binding site for Mg2+ is not formed at low (4 degrees C) temperature. PMID- 9355747 TI - Influence of thyroid hormone on the tissue-specific expression of cytochrome c oxidase isoforms during cardiac development. AB - In mammals, cytochrome c oxidase (COX) is composed of 13 different protein subunits. In the rat, two nuclear-encoded subunits, COX VIa and VIII, exist as tissue-specific isoforms: heart and liver. Using Northern-blot analysis, the levels of transcripts for the heart and liver isoforms of VIa and VIII were examined in developing rat hearts. The liver isoform was found to be the predominant form of subunit VIa and the exclusive form of VIII in the 18-day fetal hearts. The mRNA levels of the heart isoform of both subunits increased dramatically to reach adult levels by 14 days. Although the levels of the VIa- and VIII-liver isoform mRNAs remained stable throughout early development, their levels decreased by 40 and 36% respectively between the 18-day fetal stage and 18 day neonatal stage. Therefore the up-regulation of the heart isoforms and down regulation of the liver isoforms appear to be regulated in a co-ordinated manner during development. To determine if thyroid hormone influences the expression of these developmentally regulated isoforms, the RNA was also extracted from the hearts of 2-week-old hypothyroid rats. The results showed that the levels of VIII heart and VIa-liver COX mRNAs were approx. 40% lower in the hypothyroid hearts, while VIII-liver and VIa-heart COX isoform expression remained unchanged. These data demonstrate that the isoforms of COX subunits VIa and VIII are not co ordinately regulated by changes in thyroid hormone levels. Therefore we conclude that, although thyroid hormone influences the expression of isoforms, it appears to do so via a different mechanism from that which regulates the developmental transition. PMID- 9355750 TI - Common-type acylphosphatase: steady-state kinetics and leaving-group dependence. AB - A number of acyl phosphates differing in the structure of the acyl moiety (as well as in the leaving-group pKa of the acids produced in hydrolysis) have been synthesized. The Km and Vmax values for the bovine common-type acylphosphatase isoenzyme have been measured at 25 degrees C and pH 5.3. The values of kcat differ widely in relation to the different structures of the tested acyl phosphates: linear relationships between log kcat and the leaving group pKa, as well as between log kcat/Km and the leaving-group pKa, were observed. On the other hand, the Km values of the different substrates are very close to each other, suggesting that the phosphate moiety of the substrate is the main chemical group interacting with the enzyme active site in the formation of the enzyme substrate Michaelis complex. The enzyme does not catalyse transphosphorylation between substrate and concentrated nucleophilic acceptors (glycerol and methanol); nor does it catalyse H218O-inorganic phosphate oxygen exchange. It seems that no phosphoenzyme intermediate is formed in the catalytic pathway. Furthermore, during the enzymic hydrolysis of benzoyl phosphate in the presence of 18O-labelled water, only inorganic phosphate (and not benzoate) incorporates 18O, suggesting that no acyl enzyme is formed transiently. all these findings, as well as the strong dependence of kcat upon the leaving group pK1, suggest that neither a nucleophilic enzyme group nor general acid catalysis are involved in the catalytic pathway. The enzyme is competitively inhibited by Pi, but it is not inhibited by the carboxylate ions produced during substrate hydrolysis, suggesting that the last step of the catalytic process is the release of Pi. The activation energy values for the catalysed and spontaneous hydrolysis of benzoyl phosphate have been determined. PMID- 9355751 TI - Purification and characterization of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D valine synthetase from Penicillium chrysogenum. AB - delta-(L-alpha-Aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS) from Penicillium chrysogenum was purified to homogeneity by a combination of (NH4)2SO4 precipitation, protamine sulphate treatment, ion-exchange chromatography, gel filtration and hydrophobic interaction chromatography. The molecular mass of ACVS was estimated with native gradient gel electrophoresis and SDS/PAGE. The native enzyme consisted of a single polymer chain with an estimated molecular mass of 470 kDa. The denatured enzyme had an estimated molecular mass of 440 kDa. The influence of different reaction parameters such as substrates, cofactors and pH on the activity of the purified ACVS was investigated. The Km values for the three precursor substrates L-alpha-aminoadipic acid, L-cysteine and L-valine were determined as 45, 80 and 80 microM respectively, and the optimal assay concentration of ATP was found to be 5 mM (with 20 mM MgCl2). The dimer of the reaction product bis-delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (bisACV) gave feedback inhibition of the purified ACVS; the inhibition parameter KbisACV was determined as 1.4 mM. Furthermore dithiothreitol was shown to inhibit the purified ACVS. From the addition of a glucose pulse to a steady-state glucose limited continuous culture of P. chrysogenum it was found that there is glucose repression of the synthesis of ACVS and that there must be a constant turnover of ACVS owing to synthesis and degradation. PMID- 9355752 TI - Characterization and purification of a lipoxygenase inhibitor in human epidermoid carcinoma A431 cells. AB - A lipoxygenase inhibitor in the cytosolic fraction of human epidermoid carcinoma A431 cells was characterized and purified. The cytosolic inhibitor lost the inhibitory activity upon heating at 75 degrees C for 15 min or pretreating with 1 mg/ml trypsin at 37 degrees C for 60 min. Cytosol, after dialysis, lost the inhibitory activity but its inhibitory activity recovered when 1 mM GSH was added to the dialysate. The inhibitory activity of cytosol was also abolished by treatment either with 1 mM iodoacetate at 4 degrees C for 1 h or with 0.5 mM H2O2. The pI of the inhibitor was approx. 7.0. In addition to 12-lipoxygenase, the inhibitor inhibited the activities of 5-lipoxygenase and fatty acid cyclo oxygenase in a cell-free system. The inhibitor was purified by a series of column chromatographies using CM Sephadex C-50, Sephadex G-100 SF and Mono P columns. A major 22 kDa protein was obtained that was distinct from selenium-dependent glutathione peroxidase. PMID- 9355753 TI - Complete amino acid sequence of ananain and a comparison with stem bromelain and other plant cysteine proteases. AB - The amino acid sequences of ananain (EC3.4.22.31) and stem bromelain (3.4.22.32), two cysteine proteases from pineapple stem, are similar yet ananain and stem bromelain possess distinct specificities towards synthetic peptide substrates and different reactivities towards the cysteine protease inhibitors E-64 and chicken egg white cystatin. We present here the complete amino acid sequence of ananain and compare it with the reported sequences of pineapple stem bromelain, papain and chymopapain from papaya and actinidin from kiwifruit. Ananain is comprised of 216 residues with a theoretical mass of 23464 Da. This primary structure includes a sequence insert between residues 170 and 174 not present in stem bromelain or papain and a hydrophobic series of amino acids adjacent to His-157. It is possible that these sequence differences contribute to the different substrate and inhibitor specificities exhibited by ananain and stem bromelain. PMID- 9355754 TI - Co-oxidation of NADH and NADPH by a mammalian 15-lipoxygenase: inhibition of lipoxygenase activity at near-physiological NADH concentrations. AB - The purified 15-lipoxygenase from rabbit reticulocytes is capable of oxidizing NADH in the presence of linoleic acid and oxygen. This co-oxidation proceeds at a rate that amounts to approx. 7% of linoleic acid oxygenation rates. Although NADH inhibits the lipoxygenase reaction with linoleic acid as substrate (46% inhibition at 0.2 mM NADH), the reaction specificity of the enzyme was not altered since (13S)-hydroperoxy-(9Z,11E)-octadecadienoic acid was identified as the major reaction product. NADH oxidation was inhibited by NAD+ (uncompetitive with respect to linoleate and mixed/competitive with respect to NADH), and NADPH or NMNH could substitute for NADH with slightly different apparent Km values. NADH oxidation was enhanced at lower oxygen tension, but was completely prevented under anaerobic conditions. Computer-assisted modelling of 15-lipoxygenase/NADH interaction and sequence alignments of mammalian lipoxygenases with NADH dependent enzymes suggested that there is no specific binding of the coenzyme at the putative fatty acid-binding site of lipoxygenases. These results suggest that NAD(P)H might be oxidized by a radical intermediate formed during the dioxygenase cycle of the lipoxygenase reaction but that NADH oxidation might not proceed at the active site of the enzyme. The mechanism and possible biological consequences of 15-lipoxygenase-catalysed NAD(P)H oxidation are discussed. PMID- 9355755 TI - Insulin receptor/IGF-I receptor hybrids are widely distributed in mammalian tissues: quantification of individual receptor species by selective immunoprecipitation and immunoblotting. AB - The insulin receptor (IR) and type 1 insulin-like growth factor (IGF-I) receptor (IGFR) are both widely expressed in mammalian tissues, and are known to be capable of heteromeric assembly as insulin/IGF hybrid receptors, in addition to the classically described receptors. By selective immunoadsorption of radioligand/receptor complexes and by immunoblotting we have determined the fraction of insulin receptors and IGF receptors occurring as hybrids in different tissues. Microsomal membranes were isolated from tissue homogenates and solubilized with Triton X-100. Solubilized receptors were incubated with 125I-IGF I, and radioligand/receptor complexes bound by IR-specific and IGFR-specific monoclonal antibodies were quantified. The fraction of IGF-I binding sites behaving as hybrids (anti-IR-bound/anti-IGFR-bound) was approx. 40% in liver and spleen, 70% in placenta, and 85-90% in skeletal muscle and heart, similar results being obtained in rabbit and human tissues. There was no correlation between the proportion of hybrids and the ratio of 125I-insulin/125I-IGF-I binding in different tissues. The fraction of 125I-insulin bound to hybrids was too low for accurate quantification, because of the relatively low affinity of hybrids for insulin. The fraction of insulin receptors present in hybrids was therefore determined by immunoblotting. Receptors in solubilized human placental microsomal membranes were precipitated with IR-specific or IGFR-specific monoclonal antibodies, and after SDS/PAGE, blots were prepared and probed with IR-specific and IGFR-specific antisera. It was found that 15% of IR and 80% of IGFR were present in hybrids. Consistent with these figures, the overall level of IR was estimated, by blotting with the respective antibodies at concentrations shown to give equal signals with equal amounts of receptor, to be 4-fold greater than IGFR. Overall it was concluded that a significant fraction of both IR and IGFR occurs as hybrids in most mammalian tissues, including those that are recognized targets of insulin and IGF action. The fraction of hybrids in different tissues was not a simple function of the relative levels of IR and IGFR, possibly because of heterogeneity of receptor expression in different cell types. However, in placenta the proportions of IR, IGFR and hybrids were consistent with a process of random assembly reflecting the molar ratio of IR and IGFR half-receptors. PMID- 9355756 TI - Mouse white adipocytes and 3T3-L1 cells display an anomalous pattern of carnitine palmitoyltransferase (CPT) I isoform expression during differentiation. Inter tissue and inter-species expression of CPT I and CPT II enzymes. AB - The outer mitochondrial membrane enzyme carnitine palmitoyltransferase I (CPT I) represents the initial and regulated step in the beta-oxidation of fatty acids. It exists in at least two isoforms, denoted L (liver) and M (muscle) types, with very different kinetic properties and sensitivities to malonyl-CoA. Here we have examined the relative expression of the CPT I isoforms in two different models of adipocyte differentiation and in a number of rat tissues. Adipocytes from mice, hamsters and humans were also evaluated. Primary monolayer cultures of undifferentiated rat preadipocytes expressed solely L-CPT I, but significant levels of M-CPT I emerged after only 3 days of differentiation in vitro; in the mature cell M-CPT I predominated. In sharp contrast, the murine 3T3-L1 preadipocyte expressed essentially exclusively L-CPT I, both in the undifferentiated state and throughout the differentiation process in vitro. This was also true of the mature mouse white fat cell. Fully developed adipocytes from the hamster and human behaved similarly to those of the rat. Thus the mouse white fat cell differs fundamentally from those of the other species examined in terms of tis choice of a key regulatory enzyme in fatty acid metabolism. In contrast, brown adipose tissue from all three rodents displayed the same isoform profiles, each expressing overwhelmingly M-CPT I. Northern blot analysis of other rat tissues established L-CPT I as the dominant isoform not only in liver but also in kidney, lung, ovary, spleen, brain, intestine and pancreatic islets. In addition to its primacy in skeletal muscle, heart and fat, M-CPT I was also found to dominate the testis. The same inter-tissue isoform pattern (with the exception of white fat) was found in the mouse. Taken together, the data bring to light an intriguing divergence between white adipocytes of the mouse and other mammalian species. They also raise a cautionary note that should be considered in the choice of animal model used in further studies of fat cell physiology. PMID- 9355757 TI - Mechanism of cAMP-induced Ca2+ influx in Dictyostelium: role of phospholipase A2. AB - cAMP-induced Ca2+ influx in Dictyostelium follows two pathways: a G-protein dependent pathway where influx is reduced by 50-70% in Galpha2 and Gbeta-negative strains and a heterotrimeric G-protein-independent pathway. Using a pharmacological approach, we found that phospholipase A2 (PLA2) is the target of both pathways. The products of PLA2 activity, arachidonic acid (AA) and palmitic acid, induced Ca2+ influx to a similar extent as cAMP. Half-maximal activation occurred at 3 microM AA and saturation at 10 microM AA. The response to AA was quantitatively similar throughout early differentiation and thus independent of cAMP-receptor concentration. Synergy experiments revealed that cAMP and AA acted through identical pathways. The PLA2-activating peptide, a peptide with sequence similarity to the G-protein beta-subunit, activated Ca2+ influx. The G-protein independent pathway was sensitive to genistein but not to blockers of protein kinase C and other kinases, suggesting that tyrosine kinase may directly or indirectly activate PLA2 in this case. PMID- 9355758 TI - Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing platelet-activating-factor receptor lacking cytoplasmic C-terminal tail. AB - The mechanism and role of phospholipase D (PLD) activation by platelet-activating factor (PAF) were examined with Chinese hamster ovary cells stably expressing wild-type PAF receptor (WT-H cells) and truncated PAF receptor lacking the C terminal cytoplasmic tail (D-H cells). Treatment of D-H cells with PAF resulted in the rapid formation of Ins(1,4,5)P3, which was followed by a sustained phase for more than 10 min. In these cells, PAF-induced PLD activation lasted for more than 20 min. In contrast, PLD activation in WT-H cells was transient. PAF stimulation caused the biphasic formation of 1,2-diacylglycerol (DG) in both types of cell. The first phase was rapid and transient, coinciding with the Ins(1,4,5)P3 peak. The second sustained phase of DG formation was attenuated by butanol, which produces phosphatidylbutanol at the expense of phosphatidic acid (PA) by transphosphatidylation activity of PLD, and by propranolol, a selective inhibitor for PA phosphohydrolase catalysing the conversion of PA into DG. The DG level returned nearly to basal at 20 min after PAF stimulation in WT-H cells, whereas in D-H cells the elevated DG level was sustained for more than 20 min. The profile of translocation of protein kinase Calpha (PKCalpha) to membrane was similar to that of DG formation. In WT-H cells, PKCalpha was transiently associated with membranes and then returned to the cytosol. However, in D-H cells PKCalpha was rapidly translocated to and remained in membranes for more than 20 min. Butanol suppressed this sustained translocation of PKCalpha. Furthermore the mRNA levels of c-fos and c-jun by PAF in WT-H cells were much lower than those in D-H cells. Propranolol and butanol at concentrations that inhibited the formation of DG suppressed the PAF-induced mRNA expression of c-fos and c-jun. Taken together, the prolonged PLD activation in D-H cells confirmed a primary role for phospholipase C/PKC in PLD activation by PAF. Furthermore the results obtained here suggest that sustained PLD activation in turn leads to chronic activation and membrane translocation of PKCalpha, which might play an important role in the expression of c-fos and c-jun. PMID- 9355759 TI - Rapid stimulation of amyloid precursor protein release by epidermal growth factor: role of protein kinase C. AB - The amyloid precursor protein (APP) of Alzheimer's disease is a transmembrane protein that is cleaved by an uncharacterized enzyme known as alpha-secretase within its extracellular/intraluminal domain after the activation of guanine nucleotide-binding protein-coupled receptors linked to phosphoinositide hydrolysis. The secretory process results in the release of large soluble derivatives of APP (APPs), and, when elicited by muscarinic receptor activation, exhibits both protein kinase C (PKC)-dependent and tyrosine phosphorylation dependent components [Slack, Breu, Petryniak, Srivastava and Wurtman (1995) J. Biol. Chem. 270, 8337-8344]. In this report we examine the regulation of the release of APPs by epidermal growth factor (EGF) receptors, which possess intrinsic tyrosine kinase activity, and are coupled to a variety of effectors including phosphoinositide-specific phospholipase Cgamma. In A431 cells, EGF caused time-dependent and dose-dependent increases in the formation of inositol phosphates in cultures prelabelled with myo--3H-inositol, and in the release of APPs into the culture medium; the two responses exhibited similar time courses and EC50 values for EGF. Concomitant with these effects, there were concentration dependent (3-300 ng/ml) increases in the phosphorylation of tyrosine residues in several proteins, including the EGF receptor itself. The specific PKC antagonist GF 109203X decreased the effect of EGF by approx. 35% at a concentration that abolished the stimulation of the release of APPs by the PKC activator PMA. Tyrphostin AG 1478, an inhibitor of EGF receptor tyrosine kinase, abolished the EGF-induced release of APPs. These results demonstrate that in A431 cells, activation of the EGF receptor stimulates alpha-secretase activity by a mechanism that is partly dependent on PKC activity. PMID- 9355760 TI - Differential expression and regulation of ryanodine receptor and myo-inositol 1,4,5-trisphosphate receptor Ca2+ release channels in mammalian tissues and cell lines. AB - Ryanodine receptors (RyRs) and Ins(1,4,5)P3 receptors (Ins(1,4, 5)P3Rs) represent two multigene families of channel proteins that mediate the release of Ca2+ ions from intracellular stores. In the present study, the expression patterns of these channel proteins in mammalian cell lines and tissues were investigated by using isoform-specific antibodies. All cell lines examined expressed two or more Ins(1,4,5)P3R isoforms, with the type 1 Ins(1,4,5)P3R being ubiquitous. RyR isoforms were detected in only six out of eight cell lines studied. Similarly, of the nine rabbit tissues examined, RyR protein expression was detected only in brain, heart, skeletal muscle and uterus. Specific [3H]ryanodine binding was found in a number of rabbit tissues, although it was not detected in mammalian cell lines. Subcellular fractionation of SH-SY5Y human neuroblastomas revealed that the type 2 RyR and type 1 Ins(1,4,5)P3R co-localize among the fractions of a sucrose-cushion separation of crude microsomal membrane fractions. Manipulation of SH-SY5Y cells by chronic stimulation of muscarinic acetylcholine receptor (mAChR) results in a decrease in their type 1 Ins(1,4,5)P3R levels but not in the abundance of the type 2 RyR. Differentiation of these neuroblastomas by using retinoic acid did not detectably alter their expression of Ca2+-release channel proteins. Finally, differentiation of BC3H1 cells affects the expression of their Ca2+-release channel proteins in an isoform-specific manner. In summary, this study demonstrates that mammalian cell lines display distinct patterns of Ca2+ release channel protein expression. The abundance of these proteins is differentially regulated during phenotypic modifications of a cell, such as differentiation or chronic stimulation of mAChR. PMID- 9355762 TI - Stromal concentrations of coenzyme A and its esters are insufficient to account for rates of chloroplast fatty acid synthesis: evidence for substrate channelling within the chloroplast fatty acid synthase. AB - Concentrations of total CoAs in chloroplasts freshly isolated from spinach and peas were 10-20 microM, assuming a stromal volume of 66 microl per mg of chlorophyll. Acetyl-CoA and CoASH constituted at least 90% of the total CoA in freshly isolated chloroplasts. For a given chloroplast preparation, the concentration of endogenous acetyl-CoA was the same when extractions were performed using HClO4, trichloroacetic acid, propan-2-ol or chloroform/methanol, and the extracts analysed by quantitative HPLC after minimal processing. During fatty acid synthesis from acetate, concentrations of CoASH within spinach and pea chloroplasts varied from less than 0.1 to 5.0 microM. Malonyl-CoA concentrations were also very low (<0.1-3.0 microM) during fatty acid synthesis but could be calculated from radioactivity incorporated from [1-14C]acetate. Concentrations of CoASH in chloroplasts synthesizing fatty acids could be doubled in the presence of Triton X-100, suggesting that the detergent stimulates fatty acid synthesis by increasing the turnover rate of acyl-CoA. However, although taken up, exogenous CoASH (1 microM) did not stimulate fatty acid synthesis by permeabilized spinach chloroplasts. Calculated rates for acetyl-CoA synthetase, acetyl-CoA carboxylase and malonyl-CoA-acyl-carrier protein transacylase reactions at the concentrations of metabolites measured here are < 0.1-4% of the observed rates of fatty acid synthesis from acetate by isolated chloroplasts. The results suggest that CoA and its esters are probably confined within, and channelled through, the initial stages of a fatty acid synthase multienzyme complex. PMID- 9355761 TI - Human group II 14 kDa phospholipase A2 activates human platelets. AB - Recombinant human group II phospholipase A2 (sPLA2) added to human platelets in the low microg/ml range induced platelet activation, as demonstrated by measurement of platelet aggregation, thromboxane A2 generation and influx of intracellular free Ca2+ concentration and by detection of time-dependent tyrosine phosphorylation of platelet proteins. The presence of Ca2+ at low millimolar concentrations is a prerequisite for the activation of platelets by sPLA2. Mg2+ cannot replace Ca2+. Mg2+, given in addition to the necessary Ca2+, inhibits sPLA2-induced platelet activation. Pre-exposure to sPLA2 completely blocked the aggregating effect of a second dose of sPLA2. Albumin or indomethacin inhibited sPLA2-induced aggregation, similarly to the inhibition of arachidonic acid induced aggregation. Platelets pre-treated with heparitinase or phosphatidylinositol-specific phospholipase C lost their ability to aggregate in response to sPLA2, although they still responded to other agonists. This suggests that a glycophosphatidylinositol-anchored platelet-membrane heparan sulphate proteoglycan is the binding site for sPLA2 on platelets. Previous reports have stated that sPLA2 is unable to activate platelets. The inhibitory effect of albumin and Mg2+, frequently used in aggregation studies, and the fact that isolated platelets lose their responsiveness to sPLA2 relatively quickly, may explain why the platelet-activating effects of sPLA2 have not been reported earlier. PMID- 9355763 TI - Oxidation of neutrophil glutathione and protein thiols by myeloperoxidase-derived hypochlorous acid. AB - Neutrophils, when stimulated, generate reactive oxygen species including myeloperoxidase-derived HOCl. There is an associated decrease in reduced glutathione (GSH) concentration. We have shown that neutrophil GSH levels decrease on exposure to reagent HOCl, whereas the equivalent concentration of H2O2 had no effect. GSH loss occurred without cell lysis, was not reversible, and was accompanied by the loss of an equivalent proportion of the total protein thiols. No glutathione disulphide was formed. Studies with 35S-labelled cells indicated that much of the GSH lost was accounted for by mixed disulphides with protein and a product that co-migrated on HPLC with a novel compound formed in the reaction of HOCl and pure GSH. The properties of this compound are consistent with an intramolecular sulphonamide. Neutrophils stimulated with PMA lost 30-40% of their GSH and a similar proportion of protein thiols. Little glutathione disulphide was formed and the products were the same as seen with HOCl-treated cells. From the results and studies with inhibitors and scavengers, we conclude that HOCl was responsible for the GSH loss. Propargylglycine and buthionine sulphoximine, inhibitors of glutathione synthesis, enhanced GSH loss, but their effects were due to the production of long-lived chloramines that oxidized GSH with greater efficiency than HOCl, rather than to the inhibition of GSH synthesis. The lack of thiol selectivity by HOCl and irreversibility of oxidation means that GSH will provide limited antioxidant protection for thiol enzymes in stimulated neutrophils. PMID- 9355764 TI - Incorporation of copper into lysyl oxidase. AB - Lysyl oxidase is a copper-dependent enzyme involved in extracellular processing of collagens and elastin. Although it is known that copper is essential for the functional activity of the enzyme, there is little information on the incorporation of copper. In the present study we examined the insertion of copper into lysyl oxidase using 67Cu in cell-free transcription/translation assays and in normal skin fibroblast culture systems. When a full-length lysyl oxidase cDNA was used as a template for transcription/translation reactions in vitro, unprocessed prolysyl oxidase appeared to bind copper. To examine further the post translational incorporation of copper into lysyl oxidase, confluent skin fibroblasts were incubated with inhibitors of protein synthesis (cycloheximide, 10 microg/ml), glycosylation (tunicamycin, 10 microg/ml), protein secretion (brefeldin A, 10 microg/ml) and prolysyl oxidase processing (procollagen C peptidase inhibitor, 2.5 microg/ml) together with 300 microCi of carrier-free 67Cu. It was observed that protein synthesis was a prerequisite for copper incorporation, but inhibition of glycosylation by tunicamycin did not affect the secretion of 67Cu as lysyl oxidase. Brefeldin A inhibited the secretion of 67Ci labelled lysyl oxidase by 46%, but the intracellular incorporation of copper into lysyl oxidase was not affected. In addition, the inhibition of the extracellular proteolytic processing of prolysyl oxidase to lysyl oxidase had minimal effects on the secretion of protein-bound 67Cu. Our results indicate that, similar to caeruloplasmin processing [Sato and Gitlin (1991) J. Biol. Chem. 266, 5128-5134], copper is inserted into prolysyl oxidase independently of glycosylation. PMID- 9355765 TI - Analysis of the tetraspanin CD9-integrin alphaIIbbeta3 (GPIIb-IIIa) complex in platelet membranes and transfected cells. AB - The platelet integrin, alphaIIbbeta3 (GPIIb-IIIa), and the tetraspanin, CD9, are integral membrane proteins that are abundant in platelet membranes. We have identified several proteins, including CD9, which were co-precipitated by anti alphaIIbbeta3 antibody from untreated, resting platelets that were solubilized with the poly(oxyethylene) non-ionic detergent, Brij-35. Immunoblot and quantitative immunoprecipitation showed that the association of alphaIIbbeta3 with CD9 is specific and stoichiometric. The interaction between CD9 and alphaIIbbeta3 is probably hydrophobic, as Triton X-100 and hydrophobic detergents of the Brij series completely dissociated the CD9-alphaIIbbeta3 complex. Recombinant CD9 and alphaIIbbeta3 can associate after transfection into Chinese hamster ovary cells, as seen by co-immunoprecipitation and co-localization in the periphery of spreading cells and in the lamellipodia of cells plated on fibrinogen. This co-localization is absent from focal adhesions. Furthermore, anti-CD9-coated latex beads clustered alphaIIbbeta3 with CD9. This work indicates that the tetraspanin, CD9, is associated with beta3 integrins in resting platelets and transfected cells. PMID- 9355766 TI - alpha-Adrenergic stimulation induces phosphorylation of retinoblastoma protein in neonatal rat ventricular myocytes. AB - Mammalian cardiac myocytes become postmitotic shortly after birth, and the subsequent myocardial growth in adaptation to increasing workloads becomes primarily dependent on hypertrophy of existing myocytes. Although hypertrophic growth of cardiac myocytes has been extensively studied by using both in vitro and in vivo models, the molecular mechanism controlling the switch from hyperplastic to hypertrophic growth of cardiac myocytes is largely unknown. Since the majority of terminally differentiated cardiac myocytes are growth-arrested in G1/G0 phase, it has been hypothesized that the retinoblastoma protein (Rb) or its related pocket proteins which block G1/S transition becomes constitutively active during myocardial terminal differentiation. To test this hypothesis, we studied the regulation of Rb activity by alpha-adrenergic stimulation in neonatal rat ventricular myocytes which are mostly postmitotic in culture. Our results demonstrate that Rb is predominantly in the active hypo-phosphorylated state in control neonatal ventricular myocytes. alpha-Adrenergic stimulation activates G1/S transition in foetal but not neonatal rate ventricular myocytes. Although alpha-adrenergic stimulation does not activate G1/S transition in neonatal myocytes, it induces hyperphosphorylation of Rb to the same extent as in proliferating skeletal-muscle myoblasts or foetal ventricles. Hyper- but not hypo phosphorylated Rb in stimulated neonatal myocytes or proliferating skeletal muscle myoblasts fails to bind to the transcription factor, E2F, suggesting that hyper-phosphorylated Rb is inactive. Therefore F1/S transition could also be blocked at steps in addition to Rb inactivation during terminal differentiation and these blockades are refractory to alpha-adrenergic stimulation. PMID- 9355767 TI - ATP-dependent transport of bilirubin glucuronides by the multidrug resistance protein MRP1 and its hepatocyte canalicular isoform MRP2. AB - Bilirubin is secreted from the liver into bile mainly as monoglucuronosyl and bisglucuronosyl conjugates. We demonstrate for the first time that ATP-dependent transport of both bilirubin glucuronides is mediated by the multidrug resistance protein (MRP1) as well as by the distinct canalicular (apical) isoform MRP2, also termed cMRP or cMOAT (canalicular multispecific organic anion transporter). In membrane vesicles from MRP1-transfected HeLa cells mono[3H]glucuronosylbilirubin and bis[3H]glucuronosylbilirubin (each at 0.5 microM) were transported with rates of 5.3 and 3.1 pmol/min per mg of protein respectively. Rat hepatocyte canalicular membrane vesicles, which contain Mrp2 (the rat equivalent of MRP2), transported mono[3H]glucuronosylbilirubin and bis[3H]glucuronosylbilirubin at rates of 8.9 and 8.5 pmol/min per mg of protein, whereas membrane vesicles from mutant liver lacking Mrp2 showed no transport of the conjugates. In membrane vesicles from human hepatoma Hep G2 cells, which predominantly expressed MRP2, transport rates were 8.3 and 4.4 pmol/min per mg of protein for monoglucuronosylbilirubin and bisglucuronosylbilirubin respectively. ATP dependent transport of the glutathione S-conjugate -3H-leukotriene C4, an established high-affinity substrate for MRP1 and MRP2, was inhibited by both bilirubin glucuronides with IC50 values between 0.10 and 0.75 microM. The ratios of leukotriene C4 transport and bilirubin glucuronide transport, determined in the same membrane vesicle preparation, indicated substrate specificity differences between MRP1 and MRP2 with a preference of MRP2 for the glucuronides. PMID- 9355768 TI - Development of oral hygiene self-efficacy and outcome expectancy questionnaires. AB - In order to measure social cognitive constructs in the oral hygiene domain, questionnaires containing self-efficacy and outcome expectation items were developed. Items were generated to measure personal beliefs in brushing and flossing ability under a variety of circumstances, and expected outcomes from performing oral hygiene behaviors that might be positive, negative, primary and secondary. In the first study, factor scales were developed on the basis of the responses from 90 subjects awaiting dental treatment. Principal components analyses with varimax rotation revealed two self-efficacy and four outcome expectations dimensions that explained 73% and 51% of the variance, respectively. A second study that utilized 103 government employees was conducted to evaluate the psychometric properties of the questionnaires. All scales demonstrated good internal consistency and test-retest stability. Correlations with extra test measures provided preliminary evidence for the validity of the instruments. PMID- 9355769 TI - Patterns of dental caries severity in Chinese kindergarten children. AB - The dental caries status of a population group is traditionally described by mean values of decayed, missing and filled teeth or surfaces (DMFT or S). Because of the limitations of the DMF values alone, additional measures of dental caries become important. A system of describing the pattern of dental caries attack hierarchically according to severity of caries was suggested by Poulsen & Horowitz (Community Dent Oral Epidemiol 1974;2:7-11). The purpose of the present study was to analyze caries data from a group of 3-6-year-old Chinese kindergarten children according to this hierarchical system, assess the hierarchical assumptions of the system with deciduous teeth and evaluate its usefulness as an additional caries description for a kindergarten population. As part of a longitudinal field trial, baseline caries data were collected from 452 children. Caries was registered by tooth surface without the use of radiographs. Each child was assigned to one of six mutually exclusive zones of increasing caries severity, from zone 0=caries free through zone 5, the most severe, assuming that once a child was classified into a given zone it automatically belonged to all zones of lesser severity (except zone 0). On the basis of the original six zones, 61% of the children were classified correctly according to the hierarchical concept, but different alternative models which merged one or more zones together demonstrated varying percentages of correct classification, the cariologically most acceptable one placing 83% correctly. For each age group there was a close correlation between mean dmfs and increasing severity. The hierarchical model provides a valuable additional description of the caries status in deciduous teeth and is consistent with professional and epidemiological knowledge of caries attack patterns. PMID- 9355770 TI - Caries prevalence and treatment needs of 7- to 10-year-old schoolchildren in southwestern Germany. AB - The caries status of 1784 children aged 7 to 10 years was examined in a cross sectional, epidemiological study in the Rhine-Neckar-District. Results showed that 30.5% of the children had caries-free primary and 65.2% had caries-free permanent teeth. The d(m)ft index was 2.68, and the D(M)FT averaged 0.76. As in previous studies, a high risk caries group was found, with 10% of all children showing more than 50% of all carious and filled teeth in the permanent dentition. Overall, 45.6% of the children's primary teeth and 16.3% of their permanent teeth needed treatment. As indicated by higher dt:ft (DT:FT) ratios in younger age groups, dentists preferred treating older children. Apart from a higher caries prevalence in primary teeth in males, no significant sex differences were found. Children of rural origin had a higher caries experience. The results confirm previous data showing considerable improvements with a declining caries experience in the young population. But the caries status of German pre-teenage children is still moderately high according to WHO criteria. For further improvements, considerable efforts have to be made with special emphasis on prevention in high risk caries groups. PMID- 9355772 TI - A validation study of three indexes of orthodontic treatment need in the United States. AB - The purpose of this study was to compare the reliability and validity of three occlusal indexes of orthodontic treatment need in predicting the opinion of treatment need of a panel of 18 orthodontists. A set of 160 study casts representing all types of malocclusion was used. The casts were scored with the following occlusal indexes: the Index of Orthodontic Treatment Need (IOTN), the Handicapping Labio-Lingual Deviations index (HLD), and the Handicapping Malocclusion Assessment Record (HMAR modified). The diagnostic accuracy or validity of each index was calculated using the mean opinion of the orthodontic raters as a "gold standard". Receiver Operating Characteristic curves were plotted for each index. The overall diagnostic accuracy, as determined by percent area under the curve, was similar for each index: IOTN 98.6%; HLD 96.1%; HMAR 96.6%. The score optimizing the sensitivity and specificity relationship for each index was as follows: IOTN (dental health component) 4; HLD 13; HMAR 12. These results indicate that the three occlusal indexes provided valuable information for determining orthodontic treatment need. PMID- 9355771 TI - Oral health condition of 12-year-old handicapped children in Flanders (Belgium). AB - The dental condition of 626 12-year-old handicapped children with mild mental or moderate to severe mental retardation or learning impairment, being 25% of the population of each of these groups, was examined in Flanders. An evaluation of oral cleanliness showed poor oral hygiene in 31.8% of the children. No significant differences were found in oral cleanliness among types of handicapping conditions. The mean DMFT score was 2.9 (s: 2.6) and DMFS score was 5.4 (s: 5.6). Almost 21% of the children were free of caries or fillings. No significant differences were found among groups of handicapped children. Handicapped children presented a low level of restorative care (restorative index score: 48.7%). Mildly mentally retarded children demonstrated the lowest restorative index (43.9%). The caries experience of first permanent molars represented the largest part of the DMFT score (64.1%). Sealants were present in 7.9% of children examined. A considerable percentage of mildly mentally retarded children and learning impaired children did not brush daily (22.1% and 20.9%) and did not receive help with toothbrushing from their parents or carers (91.0% and 94.7%, respectively). PMID- 9355773 TI - Unmet orthodontic treatment need in rural Nigerian adolescents. AB - A survey of orthodontic treatment need was carried out among randomly selected rural Nigerian adolescents using the index of orthodontic treatment need. Altogether, 704 subjects (381 boys and 323 girls) aged 12-18 years (mean 14.8, SD 1.79) were recruited in the study. The results indicated that 12.6% of the population were in objective need of orthodontic treatment. Whilst there was a discrepancy in the proportions of Nigerian adolescents needing orthodontic treatment on aesthetic and dental health grounds, girls were found to have a more attractive dental appearance and less orthodontic treatment need than boys. However, the differences were not statistically significant (P>0.05). The correlation between the orthodontist's and the subject's rating of dental appearance was found to be low (r=0.35). The study also provided reliable baseline data for planning orthodontic services in Nigeria especially in areas where there are no dental services. PMID- 9355774 TI - Edentulousness and its rehabilitation over a 10-year period in a Finnish urban area. AB - In 1977 78, a baseline study group of 449 Finnish adults aged 30 years and over was examined in an urban area with a very high supply of dental services. The follow-up study in 1988 represents longitudinal data on 297 of these adults. In 1989 a new sample of persons aged 30-39 years was also obtained to provide cross sectional information comparable to that of the corresponding age group in the 1977-78 survey. At baseline in 1977-78, the prevalence of total tooth loss was 19.4% for adults aged 30 years and over. The corresponding figures for maxillary and mandibular edentulousness alone were 16.7% and 0.4% respectively. Ninety-four percent of totally edentulous and 89.6% of single-arch edentulous subjects were prosthetically rehabilitated. In the follow-up study, 7.7% of the originally dentate women and 6.7% of men had lost the rest of their teeth. For the new totally edentulous subjects the mean number of teeth lost was 5.7 (s 3.45), most of which were incisors. In the follow-up study, 89% of the new edentulous subjects had already been edentulous in the maxilla at baseline. Among 30-39-year olds the proportions of upper-arch and totally edentulous subjects in 1977 78/1989 were 6.7%/0.8% and 2.2%/0.8% respectively (P=0.024 for the difference between the time points). In the light of the repeated cross-sectional study, we can conclude that edentulism is very uncommon in the 30-39-year age group in this urban area. PMID- 9355775 TI - Oral cancer in the North-East of England: incidence, mortality trends and the link with material deprivation. AB - This study set out to determine the incidence of, and mortality from, oral cancer in the North-East of England between the mid-1970s and the early 1990s; to investigate its relation to material deprivation; and to test the completeness of cancer registry data. The Northern Region Cancer Registry provided details of registrations, deaths and population estimates. For analyses by deprivation, Small Area Statistics were obtained from the 1981 and 1991 censuses. In a selected sample district, 100% completeness and 89% accuracy of cancer registration of these conditions were found. For both tongue and mouth cancer, age- and sex-specific incidence and mortality rates rose with age and there was little change with time. When age-standardised registration and mortality ratios were compared between the Northern Region and England & Wales, only those for mouth cancer in males were significantly different. Crude survival from cancer of the tongue in males improved in the Northern Region from 1971-74 to 1983-86 but there was no improvement in females nor for cancer of the mouth in both sexes. In males for both tongue and mouth cancer, there was a graded increase in the standardised registration and mortality ratios from the most affluent to the most deprived areas but these differences were less marked in females. These differences in mouth cancer incidence found between England & Wales and the Northern Region probably reflect differences in lifestyles. The North ranks worst among the regions of England & Wales on a number of criteria of material deprivation, and long-term unemployment is one of the highest in the country. The analysis by deprivation has shown a clear relation to material deprivation. Whether socio-economic deprivation per se or a different risk factor behaviour associated with deprivation (smoking, alcohol consumption, poor diet) is the cause of these differences is not known. However, it is very likely that different risk factor behaviour plays a major part. PMID- 9355777 TI - Medical Treatment of ulcerative colitis: overview. PMID- 9355776 TI - A national epidemiological survey of oral mucosal lesions in Malaysia. AB - The prevalence of oral mucosal lesions in Malaysia was determined by examining a representative sample of 11,707 subjects aged 25 years and above throughout the 14 states over a period of 5 months during 1993/1994. A two-stage stratified random sampling was undertaken. A predetermined number of enumeration blocks, the smallest population unit in the census publication, was selected from each state. With the selected enumeration block, a systematic sample of living quarters was chosen with a random start. The survey instrument included a questionnaire on sociodemographic characteristics and a clinical examination. The clinical examination was carried out by 16 specially trained dental public health officers and the diagnosis calibrated with a final concordance rate of 92%. The age in the sample ranged from 25 to 115 years with a mean of 44.5+/-14.0. The sample comprised 40.2% males and 59.8% females; 55.8% were Malays, 29.4% Chinese, 10.0% Indians and 1.2% other ethnic groups. Oral mucosal lesions were detected in 1131 (9.7%) subjects, 5 (0.04%) had oral cancer, 165 (1.4%) had lesions or conditions that may be precancerous (leukoplakia, erythroplakia, submucous fibrosis and lichen planus) and 187 (1.6%) had betel chewer's mucosa. The prevalence of oral precancer was highest amongst Indians (4.0%) and other Bumiputras (the indigenous people of Sabah and Sarawak) (2.5%) while the lowest prevalence was amongst the Chinese (0.5%). PMID- 9355778 TI - The modern medical management of acute, severe ulcerative colitis. AB - The management of patients with acute, severe ulcerative colitis requires careful in-hospital assessment of the patient and the coordinated treatment of a team of experienced gastroenterologists and surgeons. Complete understanding of the potential complications and their management, especially toxic megacolon, is essential. We review the current medical arsenal and advocate a standardized approach to management that includes continuous, high dose intravenous hydrocortisone, more aggressive use of topical steroids as well as feeding the patients and continuing (but not initiating) oral 5-aminosalicylic acid (5-ASA) agents. For those patients whose disease proves refractory to intravenous steroids, intravenous cyclosporin (with an acute response rate of 82%) is an essential component in the medical management of these patients. Antibiotics should be used only when specifically indicated. Total parenteral nutrition has not been shown to be helpful in the acute setting. Air contrast barium enema and colonoscopy have been used to predict response but may be dangerous diagnostic modalities in these acutely ill patients and are no better than good clinical judgement. We review and advocate long-term management of acute response using 6 mercaptopurine or azathioprine. The surgical experience and the postoperative complications of the ileal pouch anal anastomosis, which include acute pouchitis in 50-60%, chronic pouchitis in 5-10% and recent reports of dysplasia among patients with chronic pouchitis, must be considered before colectomy is advised. Over 80% of patients with acute severe colitis can be spared colectomy using our current arsenal of medical therapies. PMID- 9355779 TI - Guidelines for the treatment of ulcerative colitis in remission. AB - The role is reviewed of sulphasalazine, 5-aminosalicylic acid (5-ASA), immunosuppressive agents and corticosteroids in the maintenance treatment of ulcerative colitis in remission. Sulphasalazine and oral 5-ASA are the drugs of first choice in preventing relapses for patients suffering from intermittent chronic ulcerative colitis. Rectally administered 5-ASA may be a valid alternative for treating patients with proctitis and left-sided ulcerative colitis. The optimal dosage of oral 5-ASA in the maintenance therapy of ulcerative colitis in remission is not clear. However, there is evidence that a higher dose of 5-ASA is more effective than low dosage in preventing relapses in patients in remission. For patients with chronically active or steroid-dependent ulcerative colitis who have achieved remission while taking immunosuppressants, continuing azathioprine or 6-mercaptopurine is indicated. Existing data cast doubts as to whether or not continuous maintenance is still necessary in patients suffering from intermittent chronic ulcerative colitis with prolonged endoscopic, clinical and histological remission. PMID- 9355780 TI - Immunosuppressive treatment for refractory ulcerative colitis--where do we stand and where are we going? AB - Effective medical treatment of ulcerative colitis is available. However, 20-40% of patients remain refractory and become steroid dependent, or have chronic activity. Azathioprine and its metabolite 6-mercaptopurine have been found to be effective in this setting, although duration of treatment and doses are not entirely clear. Neither is widely used in Europe for this indication. Methotrexate has no definitive place in the treatment of refractory colitis. Intravenous cyclosporin A induces remission in a considerable number of patients; follow-up treatment is, however, not defined. This approach may be useful to allow elective surgery. A number of other treatments have been proposed including chloroquine, interferons and anti-cytokines. None of these can currently be recommended for clinical practice. Anti-inflammatory cytokines such as interleukin-10 may be good candidates. PMID- 9355781 TI - Treatment of extraintestinal complications of ulcerative colitis. AB - Extraintestinal complications affect 25-30% of patients with ulcerative colitis. These extraintestinal disorders significantly contribute to morbidity and mortality of ulcerative colitis patients. While some disorders parallel the activity of the colitis, other abnormalities run a clinical course independent of the bowel disease. The pathogenesis of these disorders is unknown, but the variable relationships to the severity of colitis and the variable responses to a proctocolectomy suggest considerable heterogeneity. The present therapy for the various manifestations is reviewed in depth. In this respect it is important to note that colectomy should never be mandated by the extraintestinal complications of ulcerative colitis. PMID- 9355782 TI - Drug therapy for ulcerative colitis during pregnancy. AB - Women with ulcerative colitis (UC) are usually young and thus likely to undergo pregnancy. They should be advised to conceive when the disease is quiescent. Steroids and salicylates may be used normally during pregnancy, possibly without exceeding a dose of 2g/day for mesalazine. Azathioprine can be maintained if its indication is clear cut. Cyclosporin seems to be without additional risk in the pregnant woman, its use being limited to acute steroid-refractory disease, as an alternative to surgery. PMID- 9355783 TI - Assessment and management of ulcerative colitis in children. AB - The incidence of ulcerative colitis in school-age children in most parts of Europe has been steady at 1.5-2.0 per 100,000 children per year for the last 20 30 years. In comparison to adults, abdominal pain is a relatively frequent presenting symptom in children in addition to rectal bleeding, bloody diarrhoea or diarrhoea. Distribution of disease in children is generally more extensive (ratio rectal:left sided:extensive 25:30:45). There are remarkably few clinical trials of therapy in children and reasons for this are discussed. Subjective indices of disease severity and activity are unreliable in children. Objective measures such as endoscopy are of value to define the extent of ulceration and histopathological features; a test of gut protein loss using whole gut lavage gives an objective index of disease activity. Principles of medical management in children are generally the same as in adults with the additional need for scrupulous attention to nutrition and growth, and psychological factors. Reassuring results of a review of the health status of young adults who had developed ulcerative colitis in childhood are presented. Twenty-four of 27 considered themselves fully fit although nine of the patients had a permanent ileostomy. PMID- 9355784 TI - From basic science to future medical options for treatment of ulcerative colitis. AB - Both ulcerative colitis (UC) and Crohn's disease are considered the result of an unrestrained inflammatory reaction, but an explanation for the aetiopathogenesis has still not emerged. Until the predisposing and trigger factors have been clearly defined, therapeutic and preventive strategies for these disorders must, therefore, rely on interrupting or inhibiting the immunopathogenic mechanisms involved. Current therapies, such as glucocorticoids and 5-aminosalicylic acid, inhibit raised concentrations of interdependent, soluble mediators of inflammation, which may amplify one another or have parallel effects. Future medical options for treatment of UC aim at removing perpetuating antigens, blocking entry of inflammatory cells by manipulating adhesion molecules, targeting soluble mediators of inflammation by blocking proinflammatory molecules or by preserving endogenous suppressive molecules, or correcting genetic defects. It remains, however, to be determined whether targeting multi-inflammatory actions or a single key pivotal process is the better therapeutic strategy and whether subgroups of UC with different clinical courses will require different treatment approaches. PMID- 9355785 TI - Barrett's oesophagus--a ray of hope. AB - The incidence of oesophageal cancer at the cardia is believed to be increasing, especially in younger individuals. Although the causes are unknown, Barrett's oesophagus may be such a cause, since it has a proven pre-malignant potential. It is generally believed that long-standing gastro-oesophageal reflux disease predisposes to columnar transformation in the oesophagus, although it is unclear whether medical or surgical treatment has significant effects in the long term. PMID- 9355786 TI - Enkephalins and enkephalinase inhibitors in intestinal fluid and electrolyte transport. AB - Opioids have long been known to inhibit intestinal fluid and electrolyte secretion. They act locally on central and peripheral opiate receptors where they are rapidly degraded by neuropeptidases, the major one being enkephalinase (EC 3.4.24.11). A number of studies have shown that, when the problem of degradation can be overcome, enkephalins have potent antisecretory properties. In 1980, an enkephalinase inhibitor was described which increased the functional availability of enkephalins. More recently an orally active enkephalinase inhibitor, acetorphan, has been shown to inhibit infectious and chemically induced diarrhoea. Acetorphan does not appear to affect gastrointestinal motility and, although it also inhibits the breakdown of a range of other neuropeptides, such as substance P and neuropeptide Y, it is a promising agent with therapeutic potential. PMID- 9355787 TI - Incidence of Barrett's adenocarcinoma in an Italian population: an endoscopic surveillance programme. Gruppo Operativo per lo Studio delle Precancerosi Esofagee (GOSPE). AB - BACKGROUND: Barrett's oesophagus is a premalignant condition leading to adenocarcinoma. The incidence of adenocarcinoma of the oesophagus and the gastrooesophageal junction is rapidly increasing in the USA, northern and central Europe. Data from southern Europe are still unavailable. OBJECTIVE: To evaluate the incidence of oesophageal adenocarcinoma in a large cohort of Italian patients with Barrett's oesophagus. METHODS: A total of 344 patients (253 males and 91 females, age range 19-75 years) with histologically proven Barrett's oesophagus (length of metaplasia > or = 3 cm) were enrolled from November 1987 to June 1995. Endoscopic and histological examinations were scheduled at yearly intervals. RESULTS: One hundred and eighty-seven patients complied with the follow-up. The mean duration of the follow-up period was 36 months (total follow-up 562 patient years; range 12-90 months). Low grade dysplasia was found in five patients at the initial examination. During the surveillance period, dysplasia increased in frequency as well as in severity and was found exclusively in the intestinal type of Barrett's oesophagus. In all, dysplastic changes were found in seven patients (five low grade and two high grade) and adenocarcinoma developed in three patients during the follow-up. In a single case, both adenocarcinoma and specialized columnar epithelium developed without any evidence of dysplasia or intestinal metaplasia at the previous follow-up examination. This prospective study shows an incidence of adenocarcinoma in Barrett's oesophagus of 1/187 patient-years. When only patients with specialized columnar epithelium were considered, the risk of adenocarcinoma was 1/88 patient-years. CONCLUSION: The present report shows that the incidence of adenocarcinoma in Italian Barrett's oesophagus patients is in the range of that reported from other Western countries. PMID- 9355788 TI - Acetorphan prevents cholera-toxin-induced water and electrolyte secretion in the human jejunum. AB - OBJECTIVES: Acetorphan is an orally administered inhibitor of enkephalinase in the wall of the digestive tract. It prevents inactivation of endogenous opioid peptides released by submucosal and myenteric neurons. The aim of this study was to examine the effect of acetorphan on jejunal water and electrolyte transport in healthy volunteers under basal conditions and in a state of intestinal secretion induced by a bacterial enterotoxin. DESIGN: Ten volunteers in two groups were studied in an open trial. For the experimental design an intestinal perfusion technique was used. METHODS: Cholera toxin was used to induce intestinal secretion in a model employing segmental perfusion of the human proximal jejunum. Acetorphan was given orally prior to intrajejunal administration of cholera toxin; its effect on intestinal transport was measured over a period of four hours after exposure to cholera toxin. Serum levels of methylthioether of thiorphan as the main metabolite were measured throughout three experiments to assure sufficient drug absorption. RESULTS: Acetorphan had no influence on basal water and electrolyte absorption (133 vs. 140 ml/30 cm x h). In a control group with cholera toxin alone, significant water secretion was induced (131 ml/30 cm x h). Acetorphan completely prevented this secretion by leaving an absorption rate of 27 ml/30 cm x h. Intestinal electrolyte transport was also significantly changed towards absorption by acetorphan. CONCLUSION: Acetorphan can prevent jejunal water and electrolyte secretion induced by cholera toxin. Enkephalins may thus protect the small intestine from enterotoxin-induced secretion. PMID- 9355789 TI - Alterations of the phenotype of colonic macrophages in inflammatory bowel disease. AB - BACKGROUND: Intestinal macrophages play an important role in mucosal inflammation. In normal colonic mucosa we recently demonstrated a unique macrophage phenotype with attenuated immune functions. Here we present an analysis of the alterations of the phenotype of colonic macrophages in inflammatory bowel disease (IBD). METHODS: Intestinal macrophages were isolated from biopsies of patients with IBD (n =20). Flow cytometric triple fluorescence analysis was applied to study CD14, CD16, CD33, HLA-DR, CD44, CD11b, CD11c and CD3/CD19 expression. RESULTS: In IBD there was an increase in expression not only of CD14 compared to control mucosa (36.0% +/- 13.2% vs. 10.5% +/- 3.8%, P< 0.0001) but also of CD16 (28.6% +/- 10.3% vs. 10.1% +/- 3.9%, P< 0.0001), HLA-DR (53.1% +/- 15.9% vs. 27.3% +/- 9.2%, P< 0.0005), CD11b (42.8% +/- 14.2% vs. 17.4% +/- 6.8%, P< 0.0001) and CD11c (35.1% +/- 15.9% vs. 17.8% +/- 10.4%, P< 0.005.). Furthermore, a hitherto undescribed new population of macrophages could be detected by flow cytometry only in patients with ulcerative colitis (CD16++, CD11b++, CD14(low), CD33(low), CD11c-) accounting for 5.8% of all cells isolated. CONCLUSION: In contrast to colonic macrophages from normal mucosa, there is a significantly higher expression of CD14, CD16, HLA-DR, CD11b and CD11c in IBD, indicating additional macrophage populations in the inflamed mucosa. This may reflect either a recruitment of new cells from the circulation or a change in phenotype of resident cells. PMID- 9355790 TI - Gastric emptying time in patients with primary hypothyroidism. AB - BACKGROUND AND OBJECTIVES: Symptoms due to slowing of gastrointestinal motility constitute an important problem in most patients with primary hypothyroidism. The purpose of this study was to calculate gastric emptying time in patients with primary hypothyroidism using a radioisotopic method. PATIENTS AND METHODS: Fifteen hypothyroid patients (2 male, 13 female) participated in the study. Mean age was 43 (range 38-50) years. Only patients who had been recently diagnosed and had received no previous therapy were included in the study. Radiological and endoscopic examination of the upper gastrointestinal tract was carried out. Patients with gastric outlet obstruction were excluded. Twelve healthy and euthyroid people with no detectable gastrointestinal pathology were examined as a control group. The mean age of this group was 45 (range 35-55) years. Patients and control group received a semisolid meal containing 1 mCi technetium-99m sulphur colloid and measurements were made from the epigastrium. RESULTS: Mean gastric emptying half-time (t(half)) was 112.13+/-48.96 min in hypothyroid patients and 57.87+/-6.38 min in the control group. Gastric emptying time of the hypothyroid patients was significantly different from the control group (P< 0.001). There was no correlation between gastric emptying time and serum thyroid stimulating hormone (TSH) levels. CONCLUSION: We concluded that radioisotopic study is a safe, rapid and reliable method for determining gastric emptying time in patients with primary hypothyroidism. PMID- 9355791 TI - The safety of diagnostic and therapeutic ERCP as a daycase procedure with a selective admission policy. AB - OBJECTIVE: To assess if therapeutic endoscopic retrograde cholangiopancreatography (ERCP) as a daycase procedure with a selective admission policy is safe and cost-effective. DESIGN: An audit of case notes of patients who attended as a daycase for either a therapeutic or diagnostic ERCP over a 20-month period. SETTING: Stoke-on-Trent District General Hospital. PATIENTS: Case notes of all patients who had an ERCP as a daycase were audited. INTERVENTIONS: Therapeutic procedures performed as daycases included papillotomy, stent insertion, balloon dilatation or a combination of these procedures. Patients are discharged home 2 h after diagnostic or therapeutic ERCP. MAIN OUTCOME MEASURES: Thirty-day morbidity and mortality of daycase patients. RESULTS: During the 20 month period audited, 550 ERCPs were performed, of which 240 attended initially as daycases. There were 97 successful daycase therapeutic ERCPs. Ten patients were admitted immediately after ERCP including one who subsequently died from a myocardial infarction (known severe ischaemic heart disease); 87 patients were discharged 2 h after ERCP; none were admitted between 2 and 48 h after ERCP; 4 were admitted between 48 h and 30 days after ERCP with complications. There were 117 successful daycase diagnostic ERCPs; 4 patients were admitted immediately due to frailty, 4 were admitted between 2 and 48 h and 1 at 28 days after ERCP with complications. There were 24 failed diagnostic and 2 failed therapeutic ERCPs. CONCLUSION: Daycase ERCP with a selective admission policy is safe and cost effective. PMID- 9355792 TI - Eosinophilia triggered by beta-interferon therapy for chronic hepatitis C. AB - The authors describe a case of eosinophilia occurring in an atopic patient suffering from chronic active hepatitis C following a 2-week course with beta interferon. Since the most common causes of eosinophilia were ruled out by laboratory and instrumental investigations, the authors suggest a possible role of beta-interferon in triggering eosinophilia, in light of current beliefs about the factors modulating eosinophil growth, differentiation and survival. In particular, it has been supposed that the inhibitory effect of beta-interferon on gamma-interferon production could have triggered a preferential expansion of Th2 type T-helper cells whose particular profile of cytokine secretion has been shown to play a crucial role in the development of eosinophilia. To the authors' knowledge, this is the first report of eosinophilia induced by beta-interferon therapy for chronic hepatitis C. PMID- 9355794 TI - Has the time now arrived to eradicate Helicobacter pylori routinely in non-ulcer dyspepsia? PMID- 9355793 TI - Colonic tuberculosis and adenocarcinoma: an unusual presentation. AB - A case is presented of caecal tuberculosis coexisting with adenocarcinoma at the same site, unusually presenting as a right iliac fossa abscess. The relevant literature on 61 previously reported patients with coexisting tuberculosis and colonic carcinoma is reviewed. The patient was 81-years-old, female with a tender swelling in the right iliac fossa. Following examination by ultrasound, laparotomy was undertaken which revealed a large abscess cavity contaminated by Mycobacterium tuberculosis. Although anti-tuberculosis treatment was given, there was a persistent purulent discharge from the wound, so a new ultrasound, computed tomography scan and barium enema were arranged. These could not clearly differentiate between ileocaecal tuberculosis and carcinoma. A second laparotomy showed that there was an underlying adenocarcinoma. Although rare, the coexistence of colonic tuberculosis with carcinoma should be seriously considered especially in patients who fail to respond to anti-tuberculosis treatment. A definitive diagnosis can be established only by histological examination. PMID- 9355795 TI - Anaesthesia for pregnant animals. PMID- 9355796 TI - Microscopic anatomy of the ungulate placenta. PMID- 9355797 TI - Embryological development of the equine heart. PMID- 9355798 TI - Comparative aspects of fetal carbohydrate metabolism. PMID- 9355799 TI - Perinatal carbohydrate metabolism and the blood flow of the fetal liver. PMID- 9355800 TI - Fetal pulmonary development: the role of respiratory movements. AB - The lung develops before birth as a collapsible, liquid-filled, organ. Throughout the later stages of gestation the fetal lungs are maintained at a level of expansion that is considerably greater than the level achieved as a result of passive equilibration between lung recoil and the chest wall. Fetal breathing movements (FBM) are a feature of normal fetal life and, as such, are used clinically in the assessment of fetal wellbeing. By opposing lung recoil, FBM help to maintain the high level of lung expansion that is now known to be essential for normal growth and structural maturation of the fetal lungs. During 'apnoeic' periods between successive episodes of FBM, active laryngeal constriction has the effect of opposing lung recoil by resisting the escape of lung liquid via the trachea. The prolonged absence or impairment of FBM is likely to result in a reduced mean level of lung expansion which can lead to hypoplasia of the lungs. There is clinical evidence, disputed by some, that the absence of FBM exacerbates the effects of other factors that are associated with lung hypoplasia, such as premature rupture of fetal membranes and oligohydramnios. Even in the absence of such factors, prolonged or repeated reductions or abolition of FBM may contribute to impairments of fetal lung development; FBM can be inhibited by fetal hypoxaemia, hypoglycaemia, maternal alcohol consumption, maternal smoking, intra-amniotic infection and maternal consumption of sedatives or narcotic drugs. Abnormal growth of the fetal lungs has relevance for postnatal respiratory health as it is now recognised that there may be only a limited capacity after birth for the restoration of normal pulmonary architecture following impaired intra-uterine lung development. PMID- 9355801 TI - Systemic and luminal influences on the perinatal development of the gut. AB - At birth, the mammalian gastrointestinal tract (GIT) must be able to support a shift from mainly parenteral nutrition in the fetus (via the placenta) to enteral nutrition in the neonate. In the perinatal period the GIT therefore undergoes enhanced growth as well as morphological and functional differentiation, and this maturational programme is influenced by a complex interplay of local, systemic and luminal factors. This review shows how systemic and luminal factors may influence GIT development in the perinatal period of the pig and sheep, two long gestation species. Adrenocortical hormones play a pivotal role in the prepartum maturation of the GIT in addition to their better known effects on the development of many other tissues and body systems. More particularly, in the fetal pig and sheep, the prenatal development of gastric acid and gastrin secretion, and of GIT hydrolase activities (chymosin, pepsin, amylase, lactase, aminopeptidases) is influenced by cortisol. Additionally, glucocorticoids exert effects throughout the GIT by influencing morphological, cytological, and functional differentiation. Since the GIT epithelial cells comprise a renewing cell population there are also changes in cell kinetics. In addition to systemic factors, the presence of growth factors, hormones and nutrients from swallowed amniotic fluid (fetus) and colostrum (neonate) may influence GIT development. In utero, fetal fluid ingestion has been shown to modulate tissue growth, macromolecule and immunoglobulin transport, enterocyte differentiation, cell turnover and activity of brush-border hydrolases. These effects may be mediated via regulatory peptides (e.g. insulin-like growth factor I, gastrin-releasing peptides, insulin, epidermal growth factor, gastrin). A physiological role of luminally derived growth factors is supported by a number of unique structural and functional adaptations of the GIT in the fetus and neonate (low luminal proteolysis, intestinal macromolecule transport). Thus, in the pig and sheep, both systemic and luminal factors appear to play critical roles in GIT development in the perinatal period. PMID- 9355802 TI - Comparative aspects of fetal renal development. PMID- 9355803 TI - Fetal calcium homeostasis. AB - The mammalian fetus is maintained hypercalcaemic relative to its mother primarily by the action of a placental calcium pump located in the basal plasma membrane of the trophoblast. It is suggested that the activity of this pump is stimulated by a mid-molecular fragment of parathyroid hormone-related protein [PTHrP(38 94NH2)], produced in the placenta (and also in the parathyroid glands of fetal lambs and calves) as a result of post translational processing. In the sheep, calcitriol is an important determinant of fetal calcium homeostasis and it, too, stimulates the transport of calcium across the placenta. Fetal bone resorption is under the control of calcitriol, parathyroid hormone (PTH) and PTHrP. This resorption is modulated by the inhibitory effect of calcitonin. PTHrP also plays an important role in the regulation of endochondral ossification and chondrocyte maturation. PMID- 9355804 TI - Immunolocalisation of P450(C17) in the fetal sheep adrenal gland during gestation and in response to ACTH and glucocorticoid administration. AB - In sheep, increased output of cortisol from the fetal adrenal gland is critical to organ maturation and parturition. Cortisol synthesis is determined in part by the activity of P450(C17) enzyme. We have used immunohistochemistry and Western immunoblotting to examine the distribution of P450(C17) in the ovine fetal adrenal during gestation, and after ACTH or dexamethasone administration to fetuses between Days 125 and 130. The patterns were compared with changes in 3beta-hydroxysteroid dehydrogenase (3beta-HSD) localisation and levels. Adrenal tissue was obtained from four fetuses at each of Days 63-65, 100, 125-130 and term (>140 days). Further animals were chronically catheterised and infused with ACTH, dexamethasone or saline for 96 h beginning on Day 125. Immunohistochemistry for P450(C17), 3beta-HSD, and phenylethanolamine-N-methyl transferase (PNMT) was conducted using standard techniques. At Day 63-65 of pregnancy immunoreactive (ir )P450(C17) was present in cords of cells throughout the adrenal gland. Ir P450(C17) was reduced or was undetectable at Day 100, but had increased by Day 125-130, and was present throughout the zona fasciculata of the adrenal cortex of term animals. An increase in P450(C17) protein was also seen between Day 100 and 125 by Western blotting, and after ACTH treatment. Dexamethasone administration led to a marked reduction in ir-P450(C17) levels. In contrast, ir-3beta-HSD was present in the fetal adrenal cortex between Day 100 and term, and was less affected by ACTH or dexamethasone treatment. We conclude that P450(C17) in the fetal sheep adrenal is responsive to regulation by ACTH, and that changes in its levels correlate with previously reported alterations in patterns of cortisol output by the fetal adrenal gland. PMID- 9355806 TI - Development of the hypothalamus-pituitary-adrenal axis of the equine fetus: a comparative review. PMID- 9355805 TI - Catecholamines, enkephalins and the response of the fetal adrenal medulla to hypoxaemia. PMID- 9355807 TI - Stimulation of the switch in myometrial activity from contractures to contractions in the pregnant sheep and nonhuman primate. AB - The process of parturition is regulated by a set of interrelated endocrine, paracrine, and autocrine systems. Many of these systems demonstrate positive feed forward characteristics. The sequential recruitment of signals that promote the labour process demonstrates that it is not possible to attribute the designation of the factor responsible for the 'initiation of parturition' uniquely to any one signalling mechanism. For this reason we prefer to avoid the term initiation of parturition since, mechanistically, each cellular and molecular mechanism that contributes to the process is itself initiated by an earlier process. In a very real sense, the initiation of parturition can be considered to be fertilisation. Therefore we prefer to describe the key mechanisms involved as promoting, rather than initiating, the process of labour and delivery. Despite many interspecies differences, an increase in maternal plasma oestrogen in late gestation appears to play a central role in promotion of parturition. In both sheep and monkeys signals from the fetal hypothalamo-pituitary-adrenal axis increase oestrogen production by the placenta. We propose that the key fetal adrenal product is cortisol in the sheep and androgen in the monkey. Oestrogen then recruits a range of stimulators, uterotonins, that act on the prepared myometrium to initiate the switch in myometrial activity from contractures to contractions. The various mechanisms central to the process can be considered to be either, or both, activators and/or stimulators of one or more of the key terminal steps involved. PMID- 9355808 TI - Biosynthesis and possible biological roles of progestagens during equine pregnancy and in the newborn foal. AB - Major progress on the endocrinology of the pregnant mare has been possible thanks to the catheterised equine fetal preparation developed by Marian Silver. In particular, these preparations led to the identification of the source of progestagens within the feto-placental unit and provided the impetus for further work on their biosynthesis and biological activities. The biosynthesis of the progestagens involves close interaction between the fetus, the endometrium and the placenta, and gives rise to some fundamental biochemical questions. The biological role of the progestagens is also discussed: these compounds may have progestagenic activities but may also play a role in the onset of parturition. PMID- 9355809 TI - Readiness for birth: an endocrinological duet between fetal foal and mare. PMID- 9355810 TI - Identification and treatment of the compromised equine fetus: a clinical perspective. PMID- 9355811 TI - Thermoregulation and the energy requirement of the newborn foal, with reference to prematurity. PMID- 9355812 TI - Aspects of autonomic and neuroendocrine function. AB - This article provides a brief review of aspects of autonomic and neuroendocrine function studied initially in collaboration with the late Marian Silver. The importance of the sympathetic innervation to the liver in the control of glycogenolysis was established in anaesthetised animals of various species. Otherwise the work has been carried out mainly in conscious animals under strictly physiological conditions and below behavioural threshold. Investigations of the role of the autonomic innervation to the endocrine pancreas in controlling the release of pancreatic hormones, led to the realisation that the parasympathetic innervation mediates responses to glycaemic stimuli while the sympathetic innervation mediates responses to any form of stress. Studies of adrenal medullary function have confirmed that its threshold for many forms of stress is much higher than that of other components of the sympathetic system and revealed the importance of the pattern of electrical stimulation in determining the rates of release of catecholamines, enkephalins, corticotrophin-releasing factor (CRF) and adrendocorticotrophin (ACTH). The splanchnic sympathetic innervation to the adrenal cortex also plays an important role in determining glucocorticoid output by sensitising the cells to ACTH, probably mainly by the release of vasoactive intestinal peptide (VIP) from cortical nerve terminals. Finally studies of feeding in milk-fed calves have shown that suckling is associated with a remarkable hypertension and tachycardia. These cardiovascular effects are due to a selective sympathetic discharge, which does not involve the adrenal medullae, or the release of neuropeptide Y (NPY) and, at least in the calf, can be attributed to activation of adrenoceptors. PMID- 9355813 TI - Rapid cycling bipolar affective disorder: lack of relation to hypothyroidism. AB - Thyroid indices were measured after an extended period of medication-free evaluation averaging 6 weeks in 67 consecutively admitted patients with bipolar illness. Thyroid hormone levels -- thyroxine (T4), free T4 and triiodothyronine (T3) -- were not significantly different in the 31 rapid cyclers (> or = 4 affective episodes/year) than in 36 non-rapid cyclers. Analysis of covariance indicated a non-significant trend relation between higher T4 and a greater number of affective episodes in the year prior to admission and male gender when age was covaried. Several previous reports, primarily in medicated subjects, have suggested a link between rapid cycling patients and decreased peripheral thyroid indices (low hormone levels and elevated TSH), but now the majority of studies do not support such a relation. Among those in the literature, this study includes patients studied for the longest time off medications and further suggests that the commonly-cited relation between subclinical hypothyroidism and rapid cycling bipolar illness be reevaluated. PMID- 9355814 TI - Neurocognitive impairments associated with ambiguous handedness in the chronically mentally ill. AB - One form of atypical handedness, ambiguous handedness, is found in roughly one quarter of chronic schizophrenic patients. Despite its prevalence, relatively little is known about the neurocognitive underpinnings of ambiguous handedness. In the present study we examined the performance of ambiguous (n = 19) and non ambiguous (n = 39) handed chronically mentally ill inpatients on selected measures of verbal learning, motor learning and manual dexterity. The results revealed that ambiguous handers were more impaired than non-ambiguous handers in verbal learning, but not motor learning. Group differences in manual dexterity were significant for the entire sample, but not when analyses were limited to males. These findings suggest that impairments in verbal learning may be linked to the pathogenesis of ambiguous handedness in chronic psychiatric patients. PMID- 9355815 TI - Psycho-education in bipolar disorder: effect on expressed emotion. AB - In a waiting-list controlled study on a multi-family psycho-educational intervention in bipolar disorder, key relatives in the treatment group showed a significant change from high to low levels of expressed emotion (EE) compared with the control group. In addition, patients with low-EE key relatives had a significantly lower number of hospital admissions compared with those living with high-EE key relatives. The multi-family groups were well received by the participants, and there were only a few drop-outs. PMID- 9355816 TI - Family treatment, expressed emotion and relapse in recent onset schizophrenia. AB - A controlled longitudinal treatment study was carried out to investigate the effect of a behavioral family treatment on Expressed Emotion (EE) and to examine the correspondence between EE changes and relapse rates. Subjects were 52 patients with recent onset schizophrenia or related disorders and their parents. After completion of inpatient treatment they were randomly allocated to individual treatment or individual treatment plus family treatment. The family treatment consisted of education and training in communication and problem solving skills. Expressed Emotion was measured with the Five-Minute Speech Sample (FMSS). The findings show that family treatment did not have a significant positive effect on EE level. The dichotomous FMSS/EE did not systematically change and these findings were comparable with the results of prior EE research. A scoring system that included all subscores of the FMSS was somewhat more sensitive to changes. In the individual treatment condition relapse rates tended to co-occur with a change in FMSS/EE level, irrespective of the direction of this change. PMID- 9355817 TI - Expressed emotion and social function. AB - We examined whether expressed emotion (EE) influenced the social functioning of schizophrenia. Twenty-nine subjects meeting the diagnostic criteria of ICD-9 or DSM-III-R participated in the study. The Camberwell Family Interview was conducted to evaluate EE, and subjects were divided into high EE and low EE groups. The subjects had been followed up for 9 months and their social functioning was compared between the two groups as assessed with the Katz Adjustment Scales. In the high EE group, levels of both performance of socially expected activities and free-time activities slightly declined at follow-up. In contrast, those in the low EE group improved and the score increase in the level of performance of socially-expected activities was significant (P < 0.05). We confirmed the relationship of families' EE status with social functioning in schizophrenia. PMID- 9355818 TI - Aggressive behaviour in schizophrenia: role of state versus trait factors. AB - The objective of this article was to elucidate the relative importance of state vs. trait factors in determining aggressive behaviour in schizophrenia. Thirty one aggressive schizophrenia patients in rehabilitation wards were compared with 31 matched non-aggressive patients with respect to their psychopathology, phenomenologies of hallucinations and delusions, neuroleptic motor side effects, history of aggression and personality traits. Significant differences between the two groups were found in relation to psychopathology, affective responses to hallucinations/delusions, history of aggression and personality traits, but there were no significant differences regarding neuroleptic motor side effects. The effects of history of aggression as well as personality traits were independent of and similar to the total level of psychopathology, but were much smaller when compared to those of negative affective responses to hallucinations/delusions. PMID- 9355819 TI - A new method for assessing interexaminer agreement when multiple ratings are made on a single subject: applications to the assessment of neuropsychiatric symtomatology. AB - A new method is introduced for assessing levels of interexaminer agreement when multiple ratings are made on a single subject, with an application in psychiatric research. It is designed to provide an overall level of interexaminer agreement and separate indices of agreement for each examiner. These indices are based on biostatistical and clinical criteria to determine whether the ratings of any given examiner are appreciably higher or lower than the group average, or a consensus diagnosis. A number of examples, from ongoing psychiatric research, are provided to illustrate conditions favoring the application of the new methodology. Finally, the necessary software for performing the analyses is available to clinical investigators with interest in this area of assessment. PMID- 9355820 TI - A computer program for assessing interexaminer agreement when multiple ratings are made on a single subject. AB - This report describes a computer program for applying a new statistical method for determining levels of agreement, or reliability, when multiple examiners evaluate a single subject. The statistics that are performed include the following: an overall level of agreement, expressed as a percentage, that takes into account all possible levels of partial agreement; the same statistical approach for deriving a separate level of agreement of every examiner with every other examiner; and tests of the extent to which a giver examiner's rating (say a symptom score of three on a five-category ordinal rating scale) deviates from the group or overall average rating. These deviation scores are interpreted as standard Z statistics. Finally, both statistical and clinical criteria are provided to evaluate levels of interexaminer agreement. PMID- 9355821 TI - Combined liver-kidney transplantation: what are the indications? PMID- 9355822 TI - Administration of mycophenolate mofetil in a murine model of acute graft-versus host disease after bone marrow transplantation. AB - BACKGROUND: Graft-versus-host disease (GVHD) remains the most significant obstacle to the use of allogeneic bone marrow transplantation as a treatment for leukemia and other hematological malignancies. Because current GVHD treatment regimens such as cyclosporine and methotrexate are only partially effective, there is a need for new immunosuppressive drugs for the treatment of this condition. METHODS: A recently developed immunosuppressive drug, mycophenolate mofetil (MM), was tested in a fully mismatched (C57BL/6 donors to BALB/c recipients) murine model of acute GVHD after bone marrow transplantation. RESULTS: A dose regimen of 30 mg/kg/day given by oral gavage and begun at 1 day before transplant had no positive effect on survival and was found to retard the rate of marrow engraftment as measured by absolute blood neutrophil counts. In all subsequent experiments, treatment was begun on day 5 after transplant. Three different doses (30, 60, and 90 mg/kg/day) were tested, but no significant improvement in mean survival time (MST) was observed for the first two doses (P=0.412 and 0.100, respectively). The highest dose (90 mg/kg/day) reduced MST (P=0.059), and no further dose increases were attempted. MM in combination with cyclosporine also failed to improve MST compared with animals treated with cyclosporine alone or controls. CONCLUSIONS: These results suggest that MM given orally is not effective in this murine model of GVHD and may not have a role in the treatment and prevention of acute GVHD arising from bone marrow transplantation in the clinical setting. PMID- 9355823 TI - Cold ischemic injury accelerates the progression to chronic rejection in a rat cardiac allograft model. AB - BACKGROUND: The pathogenesis of chronic rejection likely involves an interplay between immunogenic and nonimmunogenic factors. The objective of this study was to determine the influence of cold ischemic preservation injury on the rate of progression to chronic rejection in the Lewis to F344 cardiac allograft model. METHODS: To induce an ischemic injury, donor hearts were stored for 3 hr at 4 degrees C in University of Wisconsin solution before transplantation. Allografts were excised at 1, 7, and 90 days after transplantation or at rejection. Vasculopathy was graded for degree of intimal thickening based on the involvement of vascular perimeter and luminal compromise. RESULTS: The degree of vessel injury in ischemic injured allografts at 90 days was significantly greater than in nonischemic injured allografts (2.8+/-0.4 vs. 1.6+/-0.5, P<0.05). Ischemic injury in syngeneic grafts did not induce a vasculopathy. Immunoperoxidase staining with R73 (anti-T cell) and ED1 (anti-macrophage) monoclonal antibodies revealed that, in ischemic injured allografts at 90 days after transplantation, the infiltrate was composed predominantly of T cells and macrophages. Additionally, ischemic injured allografts excised at 7 days after transplantation showed cellular infiltrates composed of R73-positive T cells and rare interleukin 2 receptor-positive cells, which was not observed in nonischemic allografts or ischemic syngeneic grafts. CONCLUSIONS: The progression to chronic vasculopathy in this model is principally an immunologic process, which is accelerated by an ischemic insult to the allograft. The vascular injury is mediated in part by T cells and macrophages. PMID- 9355824 TI - Role of Fas-Fas ligand interactions in the immunorejection of allogeneic mouse corneal transplants. AB - BACKGROUND: The expression of Fas ligand (FasL) in the eye has been proposed to be an important component of ocular immune privilege. Since the unusually favorable outcome of corneal transplantation is thought to result from the immune privilege of the eye, examination of the function of FasL on corneal allografts would be a test of that hypothesis. METHODS: To investigate the role of Fas-FasL interaction in corneal allografts, orthotopic corneal transplantation was performed using C57BL/6 (B6, FasL+) and B6-gld (FasL-) mice as cornea donors and BALB/c mice as recipients. The rejection rate of B6-gld grafts (FasL- group) was compared with that of normal B6 control corneas. RESULTS: The rejection rate at the final observation (8 weeks) in the FasL- group (89%) was significantly higher than in the FasL+ control group (47%). FasL expression was found on the corneal endothelium by staining with anti-FasL monoclonal antibodies. The TdT-mediated dUTP nick-end labeling assay revealed that apoptotic cells were attached to the endothelium in the control group but not in the FasL- groups. CONCLUSIONS: Apoptosis of infiltrating cells on the corneal endothelium resulting from Fas FasL interaction plays an important role in the high success rate of corneal transplantation. PMID- 9355825 TI - Cryopreservation of microencapsulated porcine pancreatic islets: in vitro and in vivo studies. AB - BACKGROUND: If the transplantation of immunoisolated porcine islets into human diabetics is to become reality, the development of a long-term storage method represents an important prerequisite. However, information on cryogenic storage of porcine islets is scanty and fragmentary. METHODS: Porcine pancreatic islets microencapsulated in alginate-polylysine-alginate membranes were cryopreserved and assessed both in vitro by static glucose challenge and in vivo in a transplantation study. Two separate methods of islet cryopreservation were compared: method A, using the Bio Cool III freezing machine, and method B, which uses the Nalgene isopropyl alcohol insulated cooler. RESULTS: Method A was found to have better preserved the ability of the microencapsulated cryopreserved islets to respond to high-glucose static challenge (7 out of 10 lots) compared with method B (1 out of 10 lots). Upon exposure to high glucose, the islet batches that did retain the ability to respond to glucose were shown to have secreted an average of 1220+/-73 pM/24 hr/islet of insulin as compared with 1528+/-118 pM/24 hr/islet for fresh islets. The presence of isobutyl methylxanthine further potentiated insulin secretion to 1805+/-81 pM/24 hr/islet and to 2410+/-104 pM/24 hr/islet for cryopreserved and free islets, respectively. Intraperitoneal transplantation of 2000 cryopreserved microencapsulated porcine islets into streptozotocin-diabetic mice resulted in the reversal of hyperglycemia in 6 out of 10 recipients for the duration of the 90-day study. CONCLUSIONS: The effective protection of the delicate porcine endocrine tissue during the cryopreservation process and the subsequent long-term storage were demonstrated with considerable success in this study. PMID- 9355826 TI - Humoral injury in porcine livers perfused with human whole blood. AB - BACKGROUND: We investigated the influence of humoral injury during xenoperfusion of porcine livers by human blood. METHODS: The porcine livers were perfused under physiological conditions for 9 hr. The perfusates consisted of porcine whole blood in group 1, human whole blood in group 2, and human whole blood with soluble complement receptor type 1 (300 microg/ml) in group 3. RESULTS: Liver enzyme release and serum hemoglobin in group 2 increased significantly after 3 hr of xenoperfusion, compared with those in group 1 and group 3 (P<0.05). Severe histological damage with minimal cellular infiltration was observed in group 2 after 6 hr of xenoperfusion, but was present only at trace levels in group 1 and group 3. In group 2, von Willebrand factor, a possible target of natural antibodies, was induced on sinusoidal endothelial cells after 3 hr of xenoperfusion, correlating with diffuse deposition of human IgM and membrane attack complex. In group 3, von Willebrand factor, human IgM, and membrane attack complex staining in the intralobular region were present at trace levels. In group 3, the indocyanine green removal capacity, representing hepatocyte function, was significantly higher than in group 2 (P<0.05). CONCLUSIONS: Based on these results, we suggest that humoral injury is a major cause of liver damage during liver xenoperfusion. The pattern of humoral injury in xenoperfused livers may be attributed to anatomical features of the liver and unique responses of sinusoidal endothelial cells to xenoperfusion. PMID- 9355827 TI - Complications and risks of living donor nephrectomy. AB - BACKGROUND: Short- and long-term patient and graft survival rates are better for living donor (vs. cadaver) kidney transplant recipients. However, donor nephrectomy is associated with at least some morbidity and mortality. We have previously estimated the mortality of living donor nephrectomy to be 0.03%. In our present study, to determine associated perioperative morbidity, we reviewed donor nephrectomies performed at our institution from January 1, 1985, to December 31, 1995. METHODS: The records of 871 donors were complete and available for review. Of these donors, 380 (44%) were male and 491 (56%) were female. The mean age at the time of donation was 38 years (range: 17-74 years), and mean postoperative stay was 4.9 days (range: 2-14 days). RESULTS: We noted two (0.2%) major complications: femoral nerve compression with resulting weakness, and a retained sponge that required reexploration. We noted 86 minor complications in 69 (8%) donors: 22 (2.4%) suspected wound infections (only 1 wound was opened), 13 (1.5%) pneumothoraces (6 required intervention, 7 resolved spontaneously), 11 (1.3%) unexplained fevers, 8 (0.9%) instances of operative blood loss > or = 750 ml (not associated with other complications), 8 (0.9%) pneumonias (all of which resolved quickly with antibiotics alone), 5 (0.6%) wound hematomas or seromas (none were opened), 4 (0.5%) phlebitic intravenous sites, 3 (0.3%) urinary tract infections, 3 (0.3%) readmissions (2 for pain control and 1 for mild confusion that resolved with discontinuation of narcotics), 3 (0.3%) cases of atelectasis, 2 (0.2%) corneal abrasions, 1 (0.1%) subacute epididymitis, 1 (0.1%) Clostridium difficile colitis, 1 (0.1%) urethral trauma from catheter placement, and 1 (0.1%) enterotomy. At our institution, no donor died or required ventilation or intensive care. We noted no myocardial infarctions, deep wound infections, or reexplorations for bleeding. Analysis, by logistic regression, identified these significant risk factors for perioperative complications: male gender (vs. female, P<0.001), pleural entry (vs. no pleural entry, P<0.004), and weight > or = 100 kg (vs. < 100 kg, P<0.02). Similar analysis identified these significant risk factors for postoperative stay > 5 days: operative duration > or = 4 hr (vs. < 4 hr, P<0.001) and age > or = 50 years (vs. < 50 years, P<0.001). CONCLUSIONS: Living donor nephrectomy can be done with little major morbidity. The risks of nephrectomy must be balanced against the better outcome for recipients of living donor transplants. PMID- 9355828 TI - Simultaneous heart and kidney transplantation as treatment for end-stage heart and kidney failure. AB - BACKGROUND: The aim of the present analysis was to define the role of simultaneous heart and kidney transplantation (HNTX) using organs from the same donor by evaluation of clinical strategy and achieved outcome compared with a reference group of concurrently single heart transplant (HTX) and kidney transplant (NTX) recipients. Compared with other organ combinations (pancreas kidney, heart-lung), HNTX has been performed infrequently and is reported mainly as case records in the literature. Because of expansion of recipient selection criteria for HTX and NTX, the number of patients requiring simultaneous replacement of both organs is increasing. METHODS: Six HNTX recipients, three of them suffering from long-standing type I diabetes, received transplants between September 1990 and March 1996 and were analyzed in terms of clinical and immunological demographics and outcome. They were compared with 379 HTX and 769 NTX recipients operated upon within this period. RESULTS: Survival for HNTX is 100% with a mean follow-up of 32.7+/-21.1 months. Cold ischemic time of the kidney was significantly shorter for HNTX than for NTX (6.5+/-1.0 hr vs. 22.1+/ 6.8 hr, P<0.005). Although HNTX patients received HLA-unmatched grafts, no rejection of the kidney has been observed to date. There was no difference for rejection of the heart in HNTX compared to HTX recipients. CONCLUSIONS: Satisfying results are obtained by HNTX and justify the use of two organs for one recipient. The favorable immunological behavior of the kidney despite use of HLA unmatched grafts is most probably explained by higher immunosuppression and short cold ischemic time, although a combination effect cannot be excluded. PMID- 9355829 TI - Metabolic factors have a major impact on kidney allograft survival. AB - BACKGROUND: At the present time, late graft loss is the major cause of kidney failure after transplantation. However, the influence of metabolic factors on this process is ill-defined. METHODS: To identify the impact of lipid metabolism, glucose metabolism, and blood pressure and their prognostic value for graft survival, data for all recipients of a kidney allograft with a potential graft survival of >15 years and a minimum graft survival of 1 month were analyzed retrospectively. Recipients of kidney grafts functioning more than 15 years (n=32) were compared with those with a graft function of less than 10 years (n=152, controls) and evaluated in a multivariate analysis. RESULTS: Low levels of serum cholesterol, triglycerides, and glucose, before and after transplantation, were accompanied by a prolonged graft survival. Prognostic factors for early graft failure included serum triglycerides >300 mg/dl, cholesterol >250 mg/dl before transplantation, serum creatinine >4.0 mg/dl 1 month after transplantation, and donor age above 45 or less than 10 years. Additionally, systolic and, particularly, diastolic blood pressure was lower in the group with a prolonged graft function as compared with controls immediately before and after transplantation. In addition, the incidence of primary graft function was lower and the incidence of acute rejection episodes higher in controls. Cold and warm ischemic time, body mass index, recipient age, and gender did not differ significantly. CONCLUSIONS: Our data suggest that metabolic parameters play an important role in the process of late graft loss after kidney transplantation. PMID- 9355830 TI - Human granulocytic ehrlichiosis in a renal transplant patient: case report and review of the literature. AB - BACKGROUND: Human ehrlichiosis, a newly described zoonotic infection, can be classified as human monocytic ehrlichiosis (HME) or human granulocytic ehrlichiosis (HGE). Although the clinical manifestations of HME and HGE are similar, the type of leukocyte infected, the etiologic agent, and the tick vector are distinct. METHODS: We report the first case of HGE in a solid organ transplant recipient and review the literature on HGE. RESULTS: Our patient displayed typical epidemiological, clinical, and laboratory features and responded promptly to therapy with doxycycline. CONCLUSIONS: Although opportunistic infections are relatively common in the posttransplant population, one must always consider other infections that occur in normal hosts as well. Human ehrlichiosis should be included in the differential diagnosis for transplant patients with fever, cytopenias, and hepatitis, especially if exposure to ticks in endemic areas has occurred. PMID- 9355831 TI - When should expanded criteria donor kidneys be used for single versus dual kidney transplants? AB - BACKGROUND: To increase the utilization of cadaveric donor kidneys, we have recently expanded our acceptable criteria to include aged donors (frequently with a history of hypertension), by selectively using both donor kidneys (dual transplant) into a single recipient. METHODS: To define when these expanded criteria donor (ECD) kidneys should be used as a single versus a dual kidney transplant, we retrospectively reviewed 52 recipients of ECD kidneys that had been turned down by all other local centers between 1/1/95 and 11/15/96. Fifteen patients received dual transplants, whereas the remaining 37 received single kidneys. Of the dual kidney recipients, 14 of 15 ECD were > or = 59 years of age, 10 of 15 were hypertensive, and 9 of 15 were both. Of the single recipients, 11 of 37 ECD were > or = 59 years of age, 11 of 37 were hypertensive, and 7 of 37 were both. All patients received cyclosporine-based triple-drug therapy. We compared seven donor (D) and sixteen recipient outcome variables in single versus dual kidney transplants as subgrouped by: (1) donor admission creatinine clearance (D-AdC(Cr)) < 90 ml/min; (2) D-age > or = 59 years; and (3) cold storage (Cld Stg) < or > 24 hr. RESULTS: In the group with D-AdC(Cr) < 90, there was a significantly higher incidence of delayed graft function (DGF) in single versus dual recipients (9 of 20 [45%] vs. 1 of 11 [9%]; P=0.04) and worse early graft function based upon mean serum creatinine at 1 and 4 weeks (5.3+/-3.3 and 2.8+/-2.0 vs. 1.7+/-0.6 and 1.4+/-0.5 mg] dl; P<0.05). In the group with D-age > or = 59, recipients of single kidneys had significantly higher mean serum creatinine at 1, 4, and 12 weeks versus recipients of dual kidneys (5.1+/-3.3, 3.4+/-2.1, 2.8+/-1.5 versus 2.8+/-2.5, 1.5+/-0.6, 1.6+/-0.5 mg/dl; P<0.05). Cld Stg time also had an impact on DGF and early outcome. Recipients of dual kidneys stored less than 24 hr had a significantly lower incidence of DGF versus single kidneys stored more than 24 hr (10% vs. 46%; P<0.05) and better early graft function based on mean serum creatinine at 1, 4, and 12 weeks (1.9+/-0.8, 1.3+/ 0.4, 1.5+/-0.2 vs. 6.6+/-3.4, 3.0+/-1.6, 2.9+/-1.9 mg/dl; P<0.05). The overall 1 year patient and graft survivals were 96% and 81% vs. 93% and 87% (P=NS) in recipients of single ECD versus dual ECD kidneys. CONCLUSIONS: In conclusion, we believe that kidneys from ECD with D-AdC(Cr) < 90 ml/min and D-age > or = 59 should be used as dual kidney transplants, keeping the Cld Stg time at < 24 hr to minimize the effect of Cld Stg on early graft function. PMID- 9355832 TI - Predictive value of host-specific donor helper T-cell precursor frequency for acute graft-versus-host disease and relapse in HLA-identical siblings receiving allogeneic bone marrow transplantation for hematological malignancies. AB - BACKGROUND: Acute graft-versus-host disease (aGVHD) is still one of the main causes of morbidity and mortality after allogeneic bone marrow transplantation. Attempts to avoid GVHD are associated with an increased risk of relapse, probably because the graft-versus-leukemia effect is also abrogated. It was recently suggested that a high frequency of host-specific donor helper T cell precursors (HTLp) might be predictive of significant aGVHD (grade > or = II). METHODS: We retrospectively studied the frequency of HTLp by means of simplified limiting dilution analysis to determine its predictive value for aGVHD and relapse. Pre bone marrow transplantation, host-specific donor HLTp frequencies were analyzed in 32 patients who had received marrow from HLA-identical siblings for hematological malignancies, in terms of aGVHD and relapse. RESULTS: HTLp frequencies were significantly higher in patients who had aGVHD > or = grade II (n=14) than in those without aGVHD (n=18) (P=0.007). Patients who relapsed (n=13) had significantly lower HTLp frequencies than those who did not relapse (n=19) (P<0.0001). The probabilities of relapse (Kaplan-Meier method) when the HTLp frequency was higher and lower than 1/200,000 were 0% and 88%, respectively (P<0.0001). CONCLUSIONS: The definition of HTLp cut-off values predictive of aGVHD and relapse should contribute to donor selection and could open the way to protocols adapting immunomodulation to the likely risk of aGVHD and relapse. PMID- 9355833 TI - Cytokine gene expression in pancreatic islet grafts in the rat. AB - We examined the production of cytokine message in allogeneic and syngeneic rat pancreatic islet grafts using specific primers and polymerase chain reaction. Freshly isolated islet preparations contained transcripts for interleukin (IL) 1alpha, IL-6, IL-10, and interferon-gamma (IFN-gamma) but not for IL-2. IL-1alpha in allogeneic grafts showed increased and consistently high expression from 1 to 7 days after transplantation, but the level in syngeneic grafts fell quickly to pretransplant levels. IL-2 and IFN-gamma transcripts were found in allogeneic grafts at 1, 3, 5, and 7 days after transplantation with a peak at day 5, but these cytokines were almost absent from syngeneic grafts. The peak of IL-6 expression was 1 day after transplantation in both syngeneic and allogeneic grafts, and then the level fell quickly. IL-10 was produced at approximately the same high level at all time points in both syngeneic and allogeneic grafts. The results show that freshly isolated islet preparations contain IL-1alpha, IL-6, IL 10, and IFN-gamma transcripts at the time of transplantation. The initial production of cytokines in islet grafts, especially IL-1, may explain phenomena such as graft nonfunction, rapid rejection, and lack of response to immunosuppression. PMID- 9355834 TI - Evidence for clonal deletion and clonal anergy after intrathymic antigen injection in a transplantation model. AB - Intrathymic (IT) antigen injection has been shown to induce antigen-specific systemic tolerance in the rodent. To delineate the mechanisms responsible for the induction of tolerance, we used the 2C line of T cell receptor transgenic mice. The majority of T cells in 2C mice express an antigen receptor specific for the major histocompatibility complex class I alloantigen Ld and can be identified with the clonotypic monoclonal antibody 1B2. IT injection of lymphoid cells expressing Ld was found to induce a significant prolongation in BALB/c skin allograft survival. The allograft prolongation was associated with a marked reduction in the number of developing 1B2+ thymocytes (clonal deletion), which occurred primarily at the CD4+ CD8+ stage of thymocyte development, as well as a reduction in the number of mature CD8+ 1B2+ 2C T cells in peripheral lymphoid tissue. In addition, CD8+ 1B2+ 2C T cells that survive deletion have decreased CD8 expression levels and a significantly reduced in vitro proliferative response to specific alloantigen (clonal anergy). Exogenous recombinant interleukin 2 restores the capacity of 2C T cells to respond in vitro to alloantigen. Experiments involving separation of cells by fluorescence-activated cell sorter indicate that there is a precise correlation between the reduction in CD8 expression and anergy induction. Collectively, these data indicate that IT antigen injection can induce antigen-specific systemic tolerance by both clonal deletion and clonal anergy. PMID- 9355835 TI - Comparative study of target antigens for primate xenoreactive natural antibodies in pig and rat endothelial cells. AB - BACKGROUND: A rat-to-primate cardiac xenograft model has been proposed as an alternative to the clinically relevant but more cumbersome pig-to-primate model for assessing the efficacy of strategies aimed at preventing xenograft hyperacute rejection. As in pig xenografts, the rejection of rat hearts was mediated by the binding of xenoreactive natural antibodies (XNA) and complement activation. The present study was conducted to identify target antigens recognized by cynomolgus and rhesus monkey IgM XNA on rat tissues and cells in comparison with pig cells. METHODS: The reactivity of rhesus or cynomolgus serum on pig and rat endothelial cells (ECs) was studied by flow cytometry, ELISA, and complement-dependent cytotoxicity, after removal of primate XNA by perfusion of pig livers, immunoadsorption on a Gal alpha(1,3)Gal affinity column, and enzymatic removal of alpha-galactosyl epitopes from the cell surface. Rat and pig EC extracts were also immunoprecipitated with primate serum and resolved in sodium dodecyl sulfate polyacrylamide gel electrophoresis. The expression of the Gal alpha(1,3)Gal epitope was analyzed on rat tissues and ECs by immunohistochemistry, flow cytometry, and Western blot, using the isolectin B4 from Griffonia simplicifolia. RESULTS: Removal of primate XNA or of alphaGal epitopes resulted in a decrease in XNA binding to pig and rat cells, leaving a similar degree of residual reactivity in the two species. At least five proteins of 260, 210, 110, 56, and 50 kDa were immunoprecipitated on rat ECs, with molecular weight similar to several proteins identified on pig ECs. These results suggest that primate XNA recognize similar antigens on rat and pig ECs. Rat cells expressed lower levels of the Gal alpha(1,3)Gal epitope than pig cells. A large proportion, but not all, of primate XNA react with this epitope on pig and rat ECs. CONCLUSION: This study suggests that the rat is a valuable species for the evaluation of genetic engineering strategies on the vascular endothelium aimed at preventing hyperacute xenograft rejection. PMID- 9355836 TI - MHC class II expression on human heart microvascular endothelial cells: exquisite sensitivity to interferon-gamma and natural killer cells. AB - BACKGROUND: Immunocytochemical analysis of human organs in situ reveals differential expression of MHC class II antigens by microvascular endothelial cells (MVEC) and endothelial cells (EC) from large vessels. In view of the role of EC as initiators of allograft rejection, it is of interest to understand the regulation of MHC class II regulation by human MVEC. We have previously isolated, cultured, and characterized MVEC from the human heart, showing that although these cells were initially MHC class II positive, the antigens were lost after about 14 days in culture. These results suggest that basal expression in vivo is maintained by circulating factors. METHODS: Here we have compared the sensitivity of human heart MVEC, human umbilical vein endothelial cells (HUVEC), and adult large vessel EC (aorta, coronary artery, and pulmonary artery) to interferon (IFN)-gamma and natural killer (NK) cell-mediated induction of MHC class II antigens. MVEC and HUVEC were cultured with 1, 5, 10, 50, 100, and 500 U/ml of IFN-gamma for 4 days, the cells were washed, and flow cytometry was used to examine HLA-DR expression at days 1, 2, 4, 7, 10, 14, and 21. EC were also cultured with purified NK cells in the presence and absence of neutralizing antibodies to IFN-gamma, and MHC class II expression was analyzed. RESULTS: As little as 5 U/ml of IFN-gamma produced 98% positive cells in heart MVEC compared with 100-500 U/ml needed for the same effect in HUVEC or other large vessel EC (coronary, aorta, pulmonary). Class II expression was maintained longer by MVEC (for 17 days) compared with HUVEC (for 10 days). NK cells and supernatant from MVEC/NK cultures induced MHC class II antigens on MVEC and HUVEC in a dose dependent fashion; the MVEC showed an enhanced sensitivity compared with the HUVEC. The NK effects were inhibited by neutralizing antibodies to IFN-gamma. The allostimulatory ability of MHC class II-positive EC was shown to be proportional to the amount of MHC class II on the cell surface. CONCLUSIONS: The results suggest that basal expression of MHC class II on human MVEC is maintained by circulating IFN-gamma and NK cells. This conclusion has implications for therapeutic interventions. PMID- 9355837 TI - Anti-CD4 monoclonal antibody-induced allograft tolerance in rats despite persistence of donor-reactive T cells. AB - Although CD4-targeted therapy abrogates acute rejection and may induce permanent graft acceptance in rodents, little is known about the mechanisms of long-term graft survival in these models. Recently, we have shown that treatment with a nondepleting anti-CD4 monoclonal antibody (mAb) (RIB-5/2) induces long-term survival of renal, heart, and skin allografts in strong major histocompatibility complex I/II incompatible rat strains. Here, we demonstrate that the development of major histocompatibility complex-specific and tissue-nonspecific tolerance rather than graft adaptation is responsible for long-term anti-CD4 mAb-induced transplant survival. Donor-specific but not third-party heart and pancreatic islet grafts were accepted permanently without adjunctive therapy in long-term kidney allograft recipients, and infusion of naive or alloimmune splenocytes failed to break the tolerant state. Interestingly, alloreactive T cells were not depleted in these long-term survivors, as ex vivo donor-specific mixed lymphocyte reaction was largely unaffected. The reverse transcriptase-polymerase chain reaction analyses of long-term renal allografts before and after donor-specific antigen challenge revealed no changes in CD3 mRNA level, but showed up-regulation of CD25, interleukin (IL) 2, interferon (IFN) gamma, IL-4, and IL-10 mRNA in the early phase, suggesting the presence of alloreactive T cells in tolerant rats. At later time points, the expression of IFN-gamma declined rapidly, whereas IL-4 persisted, resulting in a reversal of IFN-gamma/IL-4 ratio. Our data demonstrate the stability of anti-CD4 mAb-induced tolerance despite persistence of alloreactive T cells, suggesting the role of active tolerance-maintaining mechanisms. The T helper (Th) 1/Th2 shift may be involved in this regulatory process, as anti-CD4 mAb prevents acute graft-deteriorating rejection by effectively blocking Th1 responses, and well-functioning grafts may tolerize themselves by inducing regulatory cells. PMID- 9355838 TI - Studies of portal hemodynamics and hepatic oxygen consumption during acute liver allograft rejection. AB - Hemodynamics and oxygen variables, plasma cytokines, and histological features of a liver tissue sample obtained by transvenous biopsy were evaluated during 65 episodes of acute rejection. The hepatic venous pressure gradient was significantly higher in patients with acute rejection than in those without (5.1+/-0.3 vs. 3.1+/-0.2 mmHg, P<0.01). The increase in pressure gradient was related to the severity of rejection lesions. Hepatic blood flow was significantly lower in patients with than in those without acute graft rejection (1.28+/-0.11 vs. 1.75+/-0.13 L/min, P<0.05). Plasma interleukin-6 levels were significantly increased in patients with acute rejection and positively correlated with pressure gradient values. In patients with acute rejection, a significant decrease in hepatic venous oxygen content (-16%) was associated with a significant increase in hepatic oxygen consumption (+24%), whereas hepatic oxygen transport did not change significantly. In treated patients with a favorable response, the pressure gradient decreased significantly by 46%, but it remained elevated in patients who later developed chronic graft rejection. In conclusion, this study confirms that acute graft rejection may induce an increase in portal pressure, which is related to the severity of rejection lesions. It also shows that acute rejection decreases hepatic blood flow and increases hepatic oxygen consumption. In addition, it suggests that the hepatic venous pressure gradient might be useful to determine the outcome of rejection. PMID- 9355839 TI - Myofibroblast involvement in chronic transplant rejection. AB - BACKGROUND: Chronic rejection remains the major cause of late graft failure. We studied the renal tissue of 10 renal transplant patients with chronic rejection in whom biopsies had been performed at various time points over a 15-year posttransplant period to ascertain whether myofibroblasts (MF) have a role in this process. METHODS: Biopsies were grouped into five categories with respect to time after vascular anastomosis: 0 (n=10); 1 day to 3 months (n=7); 6-24 months (n=5); 36-72 months (n=7); and >72 months (n=5). A control group consisted of patients who had undergone routine biopsies at 0 (n=10), 3 (n=10), 12 (n=6), 60 (n=5), and >60 months (n=6) with no rejection. MF were identified by morphology and alpha-smooth muscle actin immunostaining. T cells and macrophages (MF) were identified using an antisera to CD3 and CD68, respectively. Collagen III deposition was similarly quantified by immunohistochemistry. Interstitial fractional area was measured by point counting. RESULTS: At all time points studied beyond time 0, there were significant increases in interstitial fractional area, collagen III staining, MF, and T-cell staining in patients with chronic rejection compared with the controls. Staining for alpha-smooth muscle actin increased with time in conjunction with worsening fibrosis and collagen deposition. CONCLUSIONS: In this study, MF were a major component of the interstitial infiltrate of the 10 patients with chronic transplant rejection. Abnormal persistence of these cells in the interstitium is one of the events that contributes to pathologic scarring of the kidney. PMID- 9355840 TI - Morphometric analysis of neointimal formation in murine cardiac grafts: III. Dissociation of intestitial fibrosis from neointimal formation. AB - BACKGROUND: This study examined the relationship between transplant vascular sclerosis (TVS) and tissue fibrosis, features of chronic rejection that can develop rapidly in accepted heterotopic murine cardiac allografts. METHODS: The rate of development of interstitial fibrosis or TVS development was determined by computerized analysis of tissue sections from DBA/2-->C57BL/6 heterotopic cardiac allografts after immunosuppression with gallium nitrate. RESULTS: In accepted cardiac allografts, neointimal fibrosis developed by 30 days after transplant, whereas TVS was minimal by day 30, and maximal by day 60. Variable levels of fibrosis were found throughout the allografts. DBA/2-->DBA/2 cardiac isografts never displayed TVS in this time period, but displayed allograft-like fibrosis within 60 days of transplantation. CONCLUSIONS: Interstitial fibrosis can be dissociated from the TVS development in this experimental model of chronic cardiac allograft rejection. Apparently, it is caused, at least in part, by alloantigen-independent factors other than TVS-related tissue ischemia. PMID- 9355841 TI - Elevated soluble interleukin 2 receptor levels found among kidney transplant recipients. AB - Although previous studies have not demonstrated a clear correlation between interleukin (IL) 2 receptor levels and immunological events after transplantation, many still suggest that these levels have clinical utility. A total of 759 serial measurements of both IL-2R and creatinine were compared over time and correlated with rejection episodes, clinical course, and immunosuppression. The profiles for the 40 patients showed several patterns, including correlation with changes in creatinine levels or with renal dysfunction, peaks in the absence of clinical findings, and discordant IL-2R and creatinine levels. Wide baseline variations in IL-2R levels confounded comparison of mean values and definition of a statistically significant rise. While tending to correlate with immunological events, elevations in IL-2R also occurred in clinically normal patients. IL-2R appears to lack specificity for immunological events. Thus, we conclude that IL-2R measurement does not have clinical diagnostic utility for monitoring renal transplant recipients. PMID- 9355842 TI - Multicentric renal cell carcinoma in a transplanted kidney. PMID- 9355843 TI - Successful allogeneic bone marrow transplantation in a 2.5-year-old boy with ongoing cytomegalovirus viremia and severe aplastic anemia after orthotopic liver transplantation for non-A, non-B, non-C hepatitis. AB - BACKGROUND: A 2.5-year-old boy received a cadaveric orthotopic liver transplant for acute liver failure due to non-A, non-B, non-C hepatitis. After transplantation, he developed thrombocytopenia and neutropenia and subsequently severe aplastic anemia. The patient also suffered from recurrent cytomegalovirus (CMV) viremia, treated with foscarnet and ganciclovir. METHODS: For treatment of his aplastic anemia, the patient underwent an allogeneic bone marrow transplantation from his HLA-identical sister after conditioning with cyclophosphamide at 200 mg/kg and antithymocyte globulin at 3 mg/kg for 5 days. Prophylactic acyclovir was given because of ongoing CMV viremia at the time of bone marrow transplantation. RESULTS: The transplant course was uneventful, with rapid engraftment. There were no signs of liver dysfunction, graft-versus-host disease, or reactivation of CMV. The patient is in excellent health, with normal liver and bone marrow function 3 years after bone marrow transplantation. CONCLUSION: This case report shows that allogeneic bone marrow transplantation is feasible and well tolerated in a patient with severe aplastic anemia after liver transplantation for acute fulminant viral hepatitis. PMID- 9355844 TI - Significance of positive cultures from donor left atrium and postpreservation fluid in heart transplantation. AB - BACKGROUND: The significance of positive perioperative cultures routinely obtained from the donor left atrium and postpreservation fluid during heart transplantation is unknown. METHODS: A retrospective chart review of 128 heart transplant recipients was done. RESULTS: A total of 106 of 128 patients had left atrial and/or postpreservation fluid cultures performed; 61 (57.5%) of them were positive. Forty-one positive left atrial or postpreservation cultures grew indolent organisms and 20 grew virulent organisms. Six donors had positive blood cultures, and five of the six did not have left atrial or postpreservation fluid cultures positive for the same organism. Seven recipients had positive blood cultures with organisms different from their corresponding left atrial or postpreservation fluid cultures. Three patients had sternal wound infections with organisms different from their donors' left atrial or postpreservation fluid cultures. Seven patients received additional antibiotics after heart transplantation specifically directed at a positive left atrial or postpreservation fluid culture for 5 to 7 days; none of them developed infection with these organisms. CONCLUSIONS: We found no evidence that positive donor left atrium or postpreservation fluid cultures increase the recipients' risk of infection. Nevertheless, we cannot refute that the small group of patients who received additional antibiotics might have developed an infection if they had not been treated. We recommend that the left atrial and postpreservation fluid cultures growing indolent organisms be discounted. However, if they grow more virulent organisms, consideration could be given to a brief course of specific therapy while awaiting recipient cultures. PMID- 9355845 TI - Apoptosis of acinar cells in pancreas allograft rejection. AB - BACKGROUND: Recently it has been recognized that apoptosis of target cells may occur during liver and kidney allograft rejection and is probably induced by infiltrating cells. Pancreas rejection is also characterized by a cellular infiltrate, however, the occurrence of apoptosis has not been investigated. We assessed whether pancreas rejection was associated with apoptosis of target cells and an influx of granzyme B (GrB)-positive or CD68-positive cells. METHODS: Eighteen pancreas biopsies (10 of 18 with rejection) from 15 patients with a pancreas-kidney transplantation were stained with the in situ end-labeling method for apoptosis, and for CD3, GrB, and CD68. RESULTS: Significantly more apoptotic acinar cells were found in biopsies with rejection when compared with biopsies without rejection. No difference was observed between the groups for GrB+ or CD68+ cells. CONCLUSION: We conclude that pancreas rejection is associated with apoptosis of acinar cells, but not with an increased influx of GrB+ cells or macrophages. PMID- 9355846 TI - Increased risk of early rejection correlates with recovery of CD3 cell count after liver transplant in patients receiving OKT3 induction. AB - BACKGROUND: We evaluated the utility of CD3 cell counts for monitoring OKT3 induction immunosuppression and for predicting early rejection in liver recipients. METHODS: In 32 adults in whom OKT3 and steroids were used to induce immunosuppression, CD3 cell subsets were labeled with CD3 (IgG1)-fluorescein isothiocyanate monoclonal antibody and assayed by flow cytometry before orthotopic liver transplantation and within 2-4 days, 5-7 days, and 8-10 days after transplantation. Trough OKT3 levels were measured at the same points in 10 patients. Early rejection (before postoperative [POD] day 21) was proven by elevated liver function tests and biopsy. Six patients were excluded for death, retransplantation, or early cessation of OKT3. RESULTS: Eight of 26 patients (30.8%) had early rejection and 18 (69.2%) had no early rejection. All had depletion of CD3 cells to <10.2% of baseline at POD 2-4. On POD 8-10, the mean CD3 count in rejectors was 213.31+/-184.98/mm3 vs. 22.71+/-32.42/mm3 in nonrejectors (P<0.001). By POD 8-10, five of eight (62.5%) patients who rejected had CD3 count recovery to >75% of baseline. No nonrejecting patient recovered to >26% of baseline (P<0.001). OKT3 levels did not correlate with CD3 recovery or rejection. CONCLUSIONS: The incidence of early rejection correlates strongly with recovery of CD3 counts by POD 10. Higher baseline CD3 counts do not predict early rejection. PMID- 9355847 TI - FK506 suppresses the mitogen-induced increase in lymphocyte adhesiveness to endothelial cells, but does not affect endothelial cell activation in response to inflammatory stimuli. AB - BACKGROUND: In vivo studies indicate that FK506 may affect endothelial cell activation. FK506 has also been reported to affect leukocyte adhesiveness and transendothelial migration. The purpose of the present study was to investigate the direct effects of FK506 on endothelial activation and on the increase in lymphocyte adhesiveness associated with mitogen stimulation. METHODS: Using flow cytometry and enzyme-linked immunosorbent assays, we studied the effects of FK506 on expression of E-selectin and intercellular adhesion molecule 1 and on the release of interleukin (IL) 6 and IL-8 from endothelial cells in response to inflammatory mediators. Expression of lymphocyte adhesion molecules and adhesion between lymphocytes and endothelial cells were also quantitated using flow cytometry. RESULTS: Endothelial activation in response to lipopolysaccharide, IL 1beta, or tumor necrosis factor-alpha was found to be unaffected by FK506. The increase in lymphocyte adhesiveness to endothelium seen after mitogen stimulation, on the other hand, was significantly depressed by FK506. The associated changes in the expression of CD11c, CD29, and CD31 were also significantly altered by FK506. CONCLUSION: In addition to its inhibitory effects on T-cell activation, FK506 may also interfere with processes leading to increased lymphocyte adhesiveness. This is thus a possible additional mechanism for its beneficial effects in connection with organ rejection and autoimmune diseases. PMID- 9355848 TI - Comment on "antilymphocyte induction therapy in cadaver renal transplantation: a retrospective multicenter United Network for Organ Sharing Study". PMID- 9355849 TI - Comment on "laparoscopic assisted live donor nephrectomy--a comparison with the open approach". PMID- 9355850 TI - The radiation environment in low-Earth orbit. AB - The radiation environment in low-Earth orbit is a complex mixture of galactic cosmic radiation, particles of trapped belts and secondary particles generated in both the spacecraft and Earth's atmosphere. Infrequently, solar energetic particles are injected into the Earth's magnetosphere and can penetrate into low Earth orbiting spacecraft. In this paper, the sources of charged-particle radiation that contribute significantly to radiation exposure on manned spacecraft are reviewed briefly, and estimates of expected dose rate for the upcoming International Space Station that are based on measurements made on the Russian Mir orbital station are provided. PMID- 9355851 TI - Carcinogens in spacecraft air. AB - The health effects of long-term exposure to ionizing radiation during spaceflight are a major concern to NASA, especially for missions beyond low-Earth orbit. Experiments involving astronauts will be conducted during the next few years to improve the risk assessment for exposures to ionizing radiation during flights; however, concomitant exposure to certain airborne chemical carcinogens during these experiments could confound the results of radiation experiments. Carcinogens can reach the spacecraft atmosphere from leaking thrusters, from off gassing materials, from chemical experiments and from human metabolism. The Johnson Space Center Toxicology Group routinely analyzes atmospheric samples collected aboard spacecraft. The exposure limits for radiomimetic compounds such as benzene are reduced because of the potential interaction between radiation and chemical exposures. Analysis of recent spacecraft air samples indicates that the following carcinogens are often present in measurable concentrations: acetaldehyde, dichloromethane, formaldehyde and isoprene. Occasionally, the carcinogens 1,2-dichloroethane, acrolein, benzene and furan are found in atmospheric samples. During normal operating conditions, the low concentrations of and limited periods of exposure to airborne carcinogens are thought to pose minimal health risks to crew members, and should not confound experiments involving sensitive methods to detect biological effects of ionizing radiation. PMID- 9355852 TI - Biodosimetry results from space flight Mir-18. AB - Astronauts are classified as radiation workers due to the presence of ionizing radiation in space. For the assessment of health risks, physical dosimetry has been indispensable. However, the change of the location of dosimeters on the crew members, the variation in dose rate with location inside the spacecraft and the unknown biological effects of microgravity can introduce significant uncertainties in estimating exposure. To circumvent such uncertainty, a study on the cytogenetic effects of space radiation in human lymphocytes was proposed and conducted for Mir-18, a 115-day mission. This study used fluorescence in situ hybridization (FISH) with whole-chromosome painting probes to score chromosomal exchanges and the Giemsa staining method to determine the frequency of dicentrics. The growth kinetics of cells and sister chromatid exchanges (SCEs) were examined to ensure that chromosomal aberrations were scored in the first mitosis and were induced primarily by space radiation. Our results showed that the frequency of chromosomal aberrations increased significantly in postflight samples compared to samples drawn prior to flight, and that the frequency of SCEs was similar for both pre- and postflight samples. Based on a dose-response curve for preflight samples exposed to gamma rays, the absorbed dose received by crew members during the mission was estimated to be about 14.75 cSv. Because the absorbed dose measured by physical dosimeters is 5.2 cGy for the entire mission, the RBE is about 2.8. PMID- 9355853 TI - Present and future radiation dosimetry for Russian cosmonauts. AB - This report describes the features of the cosmic-radiation monitoring system defined by the parameters of cosmic radiation and by the conditions of operation of dosimeters in space. The objectives of the operative and personal monitoring on board Russian spacecraft have been defined and substantiated. Accordingly, the operating characteristics of the dosimeters used during the flights of Russian manned spacecraft are given. The feasibility of perfecting the radiation monitoring facilities to be used in future manned space missions is discussed. The methods used by the Radiation Safety Service during manned space missions are described. PMID- 9355854 TI - Dose reconstruction for individuals exposed to ionizing radiation using chromosome painting. AB - To be most useful, a biomarker for dose reconstruction should employ an end point that is highly quantitative, stable with time and easily measured. Reciprocal translocations have been shown to be a promising biomarker that is linked to both prior exposure and risk, and they can be measured easily and quantitatively using fluorescence in situ hybridization. In contrast to other biomarkers that are available, the frequency of reciprocal translocations in individuals exposed to whole-body radiation is stable with time after exposure, has rather small interindividual variability and can be measured accurately at low levels of exposure. Results from recent studies demonstrate that measurements of reciprocal translocation frequencies, facilitated by chromosome painting, can be used to reconstruct radiation dose for individuals exposed in the distant past. PMID- 9355855 TI - The use of chromosomal aberrations in human lymphocytes for biological dosimetry. AB - The scoring of chromosomal aberrations in human lymphocytes provides the most sensitive method known for biological dosimetry. By scoring dicentrics in the full genome of 500 cells, average whole-body doses of about 0.1 Gy of X or gamma rays may be detected and higher doses estimated. Acute doses above about 0.2 Gy can be estimated more accurately than similar chronic doses. For radiations of higher LET, for example those encountered in the space environment, the limits of detection in grays are lower. However, expressed in sieverts, the limits of detection are more nearly independent of radiation quality. This suggests for exposure to space radiations that it may be possible to convert the yield of aberrations directly to an average whole-body dose in sieverts, which can be used as an estimate of effective dose. The scoring of translocations involving about 20% of the genome in 1000 cells using fluorescence in situ hybridization painting techniques results in a reduced sensitivity at low doses so that acute X-ray doses of about 0.3 Gy and chronic doses of about 0.4 Gy are at the limit of measurement. Better sensitivity can be achieved by scoring more cells or by using more chromosomes in color combinations, but a final limit to these approaches exists because of the higher level of spontaneous translocations than dicentrics in cells of unirradiated persons. PMID- 9355856 TI - Biodosimetry of ionizing radiation by selective painting of prematurely condensed chromosomes in human lymphocytes. AB - Painting of interphase chromosomes can be useful for biodosimetric purposes in particular cases such as radiation therapy, accidental exposure to very high radiation doses and exposure to densely ionizing radiation, for example during space missions. Biodosimetry of charged-particle radiation is analyzed in the present paper. Target cells were human peripheral blood lymphocytes irradiated in vitro with gamma rays, protons and iron ions. After exposure, lymphocytes were incubated for different times to allow repair of radiation-induced damage and then fused to mitotic hamster cells to promote premature condensation in the interphase chromosomes. Chromosome spreads were then hybridized with whole chromosome DNA probes labeled with fluorescent stains. Dose-response curves for the induction of chromatin fragments shortly after exposure, as well as the kinetics of rejoining and misrejoining, were not markedly dependent on linear energy transfer. However, after exposure to heavy ions, more aberrations were scored in the interphase cells after incubation for repair than in metaphase samples harvested at the first postirradiation mitosis. On the other hand, no significant differences were observed in the two samples after exposure to sparsely ionizing radiation. These results suggest that interphase chromosome painting can be a useful tool for biodosimetry of particle radiation. PMID- 9355857 TI - Electron paramagnetic resonance techniques and space biodosimetry. AB - This paper was presented at a workshop addressing the potential of biodosimetry techniques for use in the interplanetary space program. Some of the concerns for adequate dosimetry in space include: (1) a dosimeter that provides a permanent record of the cumulative dose and can be read independently on return to Earth; (2) a dosimeter which cannot be lost, forgotten or inadvertently removed by an individual; and (3) appropriate assessments of radiation exposures that pose an acute health risk and could jeopardize the success of an interplanetary mission. Tooth enamel is a permanent, stable biological dosimeter showing great promise in retrospective dosimetry of radiation accidents. With a proper technique, the minimum detectable dose can be in the range of tens of milligrays in extracted, prepared teeth. In addition to transient accidental doses, the cumulative dose from chronic low-level exposures (which individually may be below reportable limits) is recorded in the enamel of teeth. While many teeth remain with an individual over all or most of a lifetime, one or more are often removed due to dental problems and provide an opportunity to make dosimeteric measurements. The collection and analysis of extracted teeth in later life allows measurement of cumulative lifetime dose using the high-sensitivity techniques described in this paper. The goal of a lightweight, high-sensitivity, in vivo EPR spectrometer has not yet been realized, but its benefit to all aspects of retrospective dosimetry, terrestrial or otherwise, would be great. This paper reviews the current status of EPR dosimetry of teeth as applied to retrospective measurements of accidental exposures and outlines future research directions which will further reduce the limits of detection. PMID- 9355858 TI - Issues in cytogenetic biological dosimetry: emphasis on radiation environments in space. AB - Issues central to the reliability of cytogenetic biodosimetry are presented. Although it now appears that cytogenetic biodosimetry can be used reliably to reconstruct radiation dose after acute, uniform whole-body exposure (albeit within certain dose ranges), additional data are required to fully validate cytogenetic biodosimetry for the protracted and complex exposure conditions in space. Approaches are presented that could be used to obtain the data necessary for validation. It also appears that the use of dicentric aberrations for biodosimetry on missions lasting several months or more may not be reliable because of the large variability observed among individuals in the rate of loss of cells with dicentrics, making back-extrapolations uncertain. This may be testable, however, by comparing the results for dicentrics and translocations obtained by biodosimetry with the radiation dosimetry obtained on board the spacecraft. In addition to these and several other issues, estimates are provided of the current limitations of detection of dicentrics and reciprocal translocations. PMID- 9355859 TI - Automated biodosimetry using digital image analysis of fluorescence in situ hybridization specimens. AB - Fluorescence in situ hybridization (FISH) of metaphase chromosome spreads is valuable for monitoring the radiation dose to circulating lymphocytes. At low dose levels, the number of cells that must be examined to estimate aberration frequencies is quite large. An automated microscope that can perform this analysis autonomously on suitably prepared specimens promises to make practical the large-scale studies that will be required for biodosimetry in the future. This paper describes such an instrument that is currently under development. We use metaphase specimens in which the five largest chromosomes have been hybridized with different-colored whole-chromosome painting probes. An automated multiband fluorescence microscope locates the spreads and counts the number of chromosome components of each color. Digital image analysis is used to locate and isolate the cells, count chromosome components, and estimate the proportions of abnormal cells. Cells exhibiting more than two chromosomal fragments in any color correspond to a clastogenic event. These automatically derived counts are corrected for statistical bias and used to estimate the overall rate of chromosome breakage. Overlap of fluorophore emission spectra prohibits isolation of the different chromosomes into separate color channels. Image processing effectively isolates each fluorophore to a single monochrome image, simplifying the task of counting chromosome fragments and reducing the error in the algorithm. Using proportion estimation, we remove the bias introduced by counting errors, leaving accuracy restricted by sample size considerations alone. PMID- 9355860 TI - Radiation quality affects the efficiency of induction and the molecular spectrum of HPRT mutations in human T cells. AB - Human T lymphocytes can be used to determine the frequency and molecular spectrum of somatic cell gene mutations induced by ionizing radiations both in vivo and in vitro. In vitro exposure of these G0 cells to low-LET 137Cs gamma rays results in the induction of HPRT mutations and a predominant molecular spectrum of DNA deletions and rearrangements, particularly total gene deletions (11-12%). Similar results are found in samples from humans exposed to low-LET radiation from 131I. The doubling dose for mutation induction is calculated to be 0.8 and 1.0 Gy from these exposures performed in vitro and in vivo, respectively. In vitro studies of the effects of high-LET radiation from exposure to 222Rn also showed an induction of HPRT mutations, with a doubling dose of approximately 0.2 Gy. With this radiation, the predominant mutations were small partial deletions, with less than 2% total gene deletions. Studies of humans exposed to high-LET radiation from 239Pu showed an increased HPRT mutant frequency for the group, although no significant dosimetry could be defined. In contrast to the humans exposed to 131I, no increase in the frequency of total gene deletions was found. This is consistent with the results for 222Rn in vitro. The available data show that radiation quality affects both the efficiency of induction and the molecular spectrum of HPRT mutations in human T lymphocytes both in vitro and in vivo. The mutational spectrum may be relatively specific for radiations of different quality and thus allow a more precise measurement of the induction of somatic gene mutations resulting from individual exposures to radiation, and thereby provide more sensitive assessments of health risks. PMID- 9355861 TI - Genetic and cytogenetic markers of exposure to high-linear energy transfer radiation. AB - It has been suggested that the more closely spaced, clustered DNA breaks seen with high-LET radiations are more likely to result in chromosome intrachanges than in chromosome interchanges. We determined whether analysis of the spectra of chromosome aberrations or Hprt gene mutations in CHO-K1 cells could detect a shift to more chromosome intrachanges and therefore distinguish exposure to high LET radiation from exposure to low-LET radiation, and whether alterations in the processing of DNA breaks would influence this process. Both the frequency and type of chromosome aberrations and Hprt gene mutations were determined in CHO-K1 and xrs-5 cells exposed to 60Co gamma rays or 212Bi alpha particles. Xrs-5 cells are a radiosensitive derivative of CHO-K1 cells that are defective in rejoining of DNA double-strand breaks. The ratio of dicentrics to centric rings (F ratio) was significantly lower in CHO-K1 and xrs-5 cells exposed to alpha particles, consistent with a shift to more chromosome intrachanges with higher LET. In contrast, the frequency of large interstitial deletions at the Hprt locus, determined by multiplex polymerase chain reaction-based exon deletion analysis, was the same for both gamma-ray- and alpha-particle-exposed cells in each of the cell lines. Thus the F ratio can distinguish high-LET from low-LET radiations, and the end point is not influenced by differences in the processing of DNA double-strand breaks. The analysis of the spectrum of Hprt mutations, however, appears unable to discriminate low LET from high LET. PMID- 9355862 TI - Computer simulation of data on chromosome aberrations produced by X rays or alpha particles and detected by fluorescence in situ hybridization. AB - With fluorescence in situ hybridization (FISH), many different categories of chromosome aberrations can be recognized-dicentrics, translocations, rings and various complex aberrations such as insertions or three-way interchanges. Relative frequencies for the various aberration categories indicate mechanisms of radiation-induced damage and reflect radiation quality. Data obtained with FISH support a proximity version of the classic random breakage-and-reunion model for the formation of aberrations. A Monte Carlo computer implementation of the model, called the CAS (chromosome aberration simulator), is generalized here to high linear energy transfer (LET) and compared to published data for human cells irradiated with X rays or 238Pu alpha particles. For each kind of radiation, the CAS has two adjustable parameters: the number of interaction sites per cell nucleus and the number of reactive double-strand breaks (DSBs) per gray. Aberration frequencies for various painted chromosomes, of varying lengths, and for 11 different categories of simple or complex aberrations were simulated and compared to the data. The optimal number of interaction sites was found to be approximately 13 for X irradiation and approximately 25 for alpha-particle irradiation. The relative biological effectiveness (RBE) of alpha particles for the induction of reactive DSBs (which are a minority of all DSBs) was found to be approximately 4. The two-parameter CAS model adequately matches data for many different categories of aberrations. It can use data obtained with FISH for any one painting pattern to predict results for any other kind of painting pattern or whole-genome staining, and to estimate a suggested overall numerical damage indicator for chromosome aberration studies, the total misrejoining number. PMID- 9355863 TI - Induction of chromosome aberrations in human cells by charged particles. AB - Chromosome aberrations induced by high-energy charged particles in normal human lymphocytes and human fibroblasts have been investigated. The charged particles included 250 MeV/nucleon protons, 290 MeV/nucleon carbon ions and 1 GeV/nucleon iron ions. The energies of the charged particles were higher than in most of the studies reported in the literature. Lymphocytes were stimulated to grow immediately after irradiation, while fibroblasts were incubated at 37 degrees C for 24 h for repair. Chromosomes were collected at the first mitosis after irradiation and chromosome aberrations were scored using the fluorescence in situ hybridization (FISH) technique with a whole-chromosome 4 probe. Chromosome aberrations were classified as reciprocal exchanges, incomplete exchanges, deletions and complex exchanges. The relative biological effectiveness (RBE) for each type of aberration was calculated by dividing a dose of 4 Gy by the dose of the charged particles producing the same effect as 4 Gy of gamma rays. Results of this study showed that complex aberrations have the highest RBE for radiation of high linear energy transfer (LET) for human lymphocytes, but for fibroblasts, the greatest effect was for incomplete exchanges. For both lymphocytes and fibroblasts, iron ions induced a similar fraction of aberrant cells. PMID- 9355864 TI - Cellular effects of individual high-linear energy transfer particles and implications for tissue response at low doses. AB - The energy deposition patterns produced by the radiation environment in space can be quite different from those in conventional radiation environments. Furthermore, conventional radiation biological experiments, using randomly distributed particle tracks, cannot access some variables which may be important in determining the health effects of irradiation. Controlled microbeam irradiation provides the means to investigate the effects and unique energy deposition patterns and cell environment for a variety of end points. PMID- 9355866 TI - Pulsed-field gel electrophoretic migration of DNA broken by X irradiation during DNA synthesis: experimental results compared with Monte Carlo calculations. AB - Synchronous CHO cells were X-irradiated in G1 or mid-S phase with 30-750 Gy, and then the size distribution of DNA molecules resulting from DNA double-strand breaks (DSBs) was studied by pulsed-field gel electrophoresis (PFGE). Cells irradiated in S phase also were pulse-labeled with [3H]dThd for 15 min to compare the migration patterns of replicating DNA with those of DNA mass, measured by imaging with a CCD camera. When cells were irradiated immediately after pulse labeling, a large amount of the 3H-labeled replicating DNA was trapped in the plug, i.e. > 90% for doses < 100 Gy. As the dose increased, the percentage trapped decreased, i.e. to approximately 50% for 750 Gy. The same results were observed for DNA mass when cells were irradiated in S phase, except that much less of the DNA was trapped, i.e. approximately 60% for 70-100 Gy, which produced approximately 2-Mbp molecules, compared to approximately 10% for 750 Gy, which produced approximately 0.3-Mbp molecules. These results and the migration patterns of DNA released into the lane indicated that large molecules are trapped more readily than small molecules because they contain more replicating regions (bands with bubbles) of DNA than small molecules. Our interpretation is that as the dose increases, a greater fraction of the breaks occur between the replicating bands, thus releasing linear molecules that are not replicating. The relatively small amount of 3H-labeled replicating DNA that is released from the PFGE plug migrates aberrantly, with a small amount migrating like linear G1-phase molecules and a large amount, depending on dose, migrating much more slowly than the DNA mass from cells irradiated in G1 or S phase. To explain these results, a Monte Carlo computer program was written to introduce DSBs randomly into DNA that is configured according to a model of DNA replication that is developed in a related study (Dewey and Albright, Radiat. Res. 148, 421-434, 1997). In relating the experimental observations to the results of the Monte Carlo calculations, we assumed that (a) molecules containing replication bubbles with and without forks are trapped in the PFGE plug, (b) linear molecules and molecules with replication forks only that are < or = 8 Mbp are released into the lane, and (c) molecules having replication forks migrate more slowly than linear molecules. PMID- 9355865 TI - Regional gene mapping using mixed radiation hybrids and reverse chromosome painting. AB - We describe a new approach for low-resolution physical mapping using pooled DNA probe from mixed (non-clonal) populations of human-CHO cell hybrids and reverse chromosome painting. This mapping method is based on a process in which the human chromosome fragments bearing a complementing gene were selectively retained in a large non-clonal population of CHO-human hybrid cells during a series of 12- to 15-Gy gamma irradiations each followed by continuous growth selection. The location of the gene could then be identified by reverse chromosome painting on normal human metaphase spreads using biotinylated DNA from this population of "enriched" hybrid cells. We tested the validity of this method by correctly mapping the complementing human HPRT gene, whose location is well established. We then demonstrated the method's usefulness by mapping the chromosome location of a human gene which complemented the defect responsible for the hypersensitivity to ionizing radiation in CHO irs-20 cells. This method represents an efficient alternative to conventional concordance analysis in somatic cell hybrids where detailed chromosome analysis of numerous hybrid clones is necessary. Using this approach, it is possible to localize a gene for which there is no prior sequence or linkage information to a subchromosomal region, thus facilitating association with known mapping landmarks (e.g. RFLP, YAC or STS contigs) for higher resolution mapping. PMID- 9355867 TI - Developing a model of DNA replication to be used for Monte Carlo calculations that predict the sizes and shapes of molecules resulting from DNA double-strand breaks induced by X irradiation during DNA synthesis. AB - A Monte Carlo computer program was written to introduce double-strand breaks (DSBs) randomly into cellular DNA that is configured according to different models of DNA replication. Then, from a review of the literature using DNA fiber autoradiography and other studies relating to rates of replication of DNA that is organized in approximately 3-Mbp regions or bands, a particular model for DNA replication was developed. Using this model, Monte Carlo calculations were made to predict the types and sizes of molecules that would result from introducing DSBs into DNA when synchronous cells are irradiated in the middle of S phase. Then results of the Monte Carlo calculations were compared with migration profiles obtained by pulsed-field gel electrophoresis (PFGE) for molecular size distributions of linear DNA molecules. For these comparisons, CHO cells irradiated in S phase also were pulse-labeled at the time of irradiation with [3H]dThd for 15 min to compare the migration patterns of 3H-labeled replicating DNA with those of the mass of S-phase DNA, measured by imaging with a CCD camera. For the Monte Carlo calculations, we assumed from the reports in the literature that molecules containing replication bubbles with and without forks would be trapped in the PFGE plug. We also assumed that those molecules that are < or = 8 Mbp, both linear and with replication forks, would be released into the lane. However, approximately 75% of the 3H-labeled DNA that is released from the plug migrated much more slowly than linear molecules, which we attributed to the slow migration of 3H-labeled molecules having replication forks not attached to bubbles. The percentages of both mass of S-phase DNA and 3H-labeled replicating DNA released from the plug, as determined by PFGE, were compared with comparable values determined from Monte Carlo calculations. A DNA replication model that provides good agreement between the PFGE results and Monte Carlo calculations is described. Furthermore, Monte Carlo methodology is presented that can be used for comparing data obtained with PFGE with results of Monte Carlo calculations that are based on different models of DNA replication and different assumptions for the migration of various types of replicating molecules. PMID- 9355868 TI - Low-dose radiation sensitivity and induced radioresistance to cell killing in HT 29 cells is distinct from the "adaptive response" and cannot be explained by a subpopulation of sensitive cells. AB - Several reports using two different improved assays of clonogenicity have indicated the presence of a hypersensitive region in the radiation survival response at low doses, followed by an increase in radioresistance, in many mammalian cell lines. Mathematical modeling of these responses has suggested that it is unlikely that this effect can be explained by the presence of a small subpopulation of sensitive cells; however, this possibility cannot be excluded solely on the basis of those results. A second explanation has been offered which hypothesizes that a radiation-induced mechanism causes an increase in cellular radioresistance. This proposal has led to speculation that the substructure observed at low doses in these cell lines is related to the adaptive response, which hypothesizes the induction of a repair mechanism after a small priming dose of radiation which can protect cells against a larger second dose given several hours later. We have investigated these proposals with a study of priming doses using human tumor HT-29 cells, which we have previously shown to exhibit low-dose hyper-radiosensitivity. Our results provide significant evidence that this effect cannot be explained by a subpopulation of sensitive cells. However, the results also suggest that the radiation-induced increase in radioresistance observed in this cell line is distinct from the adaptive response. PMID- 9355869 TI - Sensitization of renal carcinoma to radiation using alpha interferon (IFNA) gene transfection. AB - The rationale for this study was that local delivery of interferon-alpha (IFN alpha) by gene transfection may be of value during radiotherapy. To investigate the feasibility of this approach, cells of the human renal carcinoma cell line R11 were transfected with the IFNA gene and evaluated for radiation responses in vitro by clonogenic assays. R11 cells expressing IFN-alpha after gene transfection were more sensitive to radiation than R11 control cells (SF2 = 0.33 and 0.51, respectively). In addition to increasing radiosensitivity, IFNA gene transfection slowed cellular growth and reduced the plating efficiency in clonogenic assays. The addition of exogenous rhIFN-alpha to cells at different times relative to irradiation showed that its presence during the postirradiation period was critical for radiosensitization, but repair of sublethal damage did not seem to be affected. No apoptosis of R11 cells was found 1-5 days after exposure to 2-25 Gy with or without IFN-alpha. Extensive formation of multinuclear giant cells was present beginning 2 days after irradiation; however, IFN-alpha did not cause any major alterations in the yield of radiation-induced giant cells. These studies suggest that gene transfection might be an effective means of delivering IFN-alpha for clinical use in radiotherapy of cancer. PMID- 9355870 TI - Dependence of induction of interphase death of Chinese hamster ovary cells exposed to accelerated heavy ions on linear energy transfer. AB - Induction of interphase death was examined in Chinese hamster ovary cells exposed to accelerated heavy ions (carbon, neon, argon and iron) of various linear energy transfers (LETs) (10-2000 keV/microm). The fraction of cells that underwent interphase death was determined by observing individual cells with time-lapse photography (direct method) as well as by counting cells undergoing interphase death made visible by the addition of caffeine (indirect method). After exposure to X rays, interphase death increased linearly with dose above a threshold of about 10 Gy, whereas it increased at a higher rate without a threshold after exposure to high-LET heavy ions. The relative biological effectiveness (RBE) compared to X rays, as determined at the 50% level of induction, increased with LET, reached a maximum at an LET of approximately 230 keV/microm and then decreased with further increase in LET. The range of LET values corresponding to the maximum RBE appears to be narrower for interphase death than for reproductive death (120-230 keV/microm), as assayed using loss of colony-forming ability as a criterion. The inactivation cross section for interphase cell death reached a plateau of 5-10 microm2. This means that the probability for the induction of interphase death by traversal of a single heavy-ion track through the nucleus (size: about 130 microm2) is about 0.04-0.08. PMID- 9355871 TI - A comparison of the modes and kinetics of heat-induced cell killing in HeLa and L5178Y cells. AB - The mode and kinetics of cell killing in HeLa and L5178Y cells were investigated to elucidate possible relationships between the mechanisms and modes of heat induced cell death. L5178Y cells were heat-shocked for either 24 min at 43 degrees C or 30 min at 45 degrees C, while HeLa cells were given only the latter treatment. The degree of heat-induced excess nuclear protein correlated with cell death in HeLa cells but not in L5178Y cells. This difference suggests that the mechanism of cell death differs between these cell lines. With L5178Y cells the main mode of death after treatment at 43 degrees C was apoptosis with little progression of cells through the cell cycle. However, after treatment at 45 degrees C these cells died by necrosis without progressing through the cell cycle. HeLa cells were found to die by a mechanism other than apoptosis after significant progression of cells through the cell cycle and perturbation of the normal distribution of cells in the phases of the cell cycle (specifically, the fraction of cells in S and G2 phase increased 50% and 30%, respectively, prior to the occurrence of measurable cell death). These results are consistent with the hypothesis that the response to injury which has the potential to be lethal varies between different cell types, and results in the induction of different pathways leading to cell death. PMID- 9355872 TI - A study of the effects of exposure on cleanup workers at the Chernobyl nuclear reactor accident using multiple end points. AB - Blood samples were collected from 192 exposed workers who participated in the cleanup after the April 26, 1986, nuclear reactor accident at Chernobyl, Ukraine. These samples, together with samples from 73 individuals living in Russia but not involved in Chernobyl cleanup activities, were collected during September 1991 to May 1996 and shipped to the U.S. for evaluation by three bioassays: cytogenetic analysis based on chromosome painting, HPRT mutation analysis and glycophorin A (GPA) variant analysis. Univariate statistical analyses of the results of each bioassay (including adjustments for age, smoking status and estimated precision of the bioassay) found greater frequencies of chromosome translocations and HPRT mutant T lymphocytes among the exposed individuals compared to the controls (P < or = 0.01). GPA analyses showed no significant difference for exposed compared to controls for either hemizygous, N/O, or homozygous, N/N, variant cell frequency. Multivariate analysis of variance of the subset of 44 exposed and 14 unexposed individuals with measurements from all three bioassays found elevated frequencies of chromosomal translocations and HPRT mutants, and reduced frequencies for both GPA end points among the exposed persons compared to the controls. However, none of these differences, considered singly or in combination, was statistically significant (although statistical power is low due to small sample sizes). Mean estimated dose, based on cytogenetic response, for those exposed was 9 cGy (range 0 to 51 cGy) and was less than that estimated by physical dosimetry (25 cGy). Correlation between the end points of the bioassays and estimated physical dosimetry was low (r < 0.2); the only significant correlation found was for physical dose estimate and dates worked at Chernobyl (r = 0.4, P < 0.01), with those working soon after the accident receiving greater estimated doses. PMID- 9355873 TI - Interactive lethal and mutagenic effects of ultraviolet light and bleomycin in yeast: synergism or antagonism? AB - The mutagenic interactions of ultraviolet light and bleomycin in haploid populations of Saccharomyces cerevisiae were analyzed. Survival and mutation frequency as a function of different bleomycin concentrations after one conditioning dose of UV radiation were determined. Furthermore, corresponding interaction functions and sensitization factors were calculated. A synergistic interaction between UV light and bleomycin was shown for both lethal and mutagenic events when the cells were in nutrient broth during the treatments. Conversely, the interaction between UV light and bleomycin was antagonistic when the cells were in deionized water during the treatment. The magnitude of lethal and mutagenic interactions depends on dose, and thus presumably on the number of lesions. The observed interactions between UV light and bleomycin suggest that the mechanism that is most likely involved is the induction of repair systems with different error probabilities during the delay of cell division. PMID- 9355874 TI - Radiation damage to DNA: techniques, quantitation and mechanisms. Bowness-on Windermere, Lake District, United Kingdom, April 19-24, 1997. PMID- 9355875 TI - Is hyperbaric oxygen more effective than carbogen/nicotinamide in tumor radiation response? PMID- 9355876 TI - Comments on hyperbaric oxygen and carbogen/nicotinamide with fractionated radiation. PMID- 9355877 TI - News from the FDA Cardio-renal Advisory Committee meeting of June 26, 1997. PMID- 9355878 TI - Minimally invasive heart surgery. PMID- 9355879 TI - Lipoprotein(a): molecular mischief in the microvasculature. PMID- 9355880 TI - Monitoring tissue repair and fibrosis from a distance. PMID- 9355881 TI - Nitrovasodilators have (small) direct effects on cardiac contractility: is this important? PMID- 9355882 TI - Biochemical markers of myocardial injury in children. PMID- 9355883 TI - A new treatment for severe pulmonary embolism: percutaneous rheolytic thrombectomy. AB - BACKGROUND: The rheolytic thrombectomy catheter has been specially designed to remove intravascular thrombus from coronary and peripheral arteries. It demonstrates a practical application of Bernoulli's principle relating to a low pressure zone in the region of a high-velocity jet. In this device, this effect is created by direct high-pressure saline jets located at the tip. Thrombus is drawn into this region and, because of the large pressure difference, undergoes mechanical thrombolysis due to the powerful mixing forces. The resulting microparticles are aspirated through the same catheter and removed from the body. METHODS AND RESULTS: We report the use of this device in two patients presenting with severe pulmonary embolism and contraindications to thrombolytic therapy. The two procedures were successfully performed with an excellent immediate angiographic result at the site of the rheolytic thrombectomy. In both cases, the clinical improvement was maintained at follow-up with the same good angiographic result and a decrease to a normal level of the systolic pulmonary pressure. CONCLUSIONS: This preliminary results suggest that this easy technical method may be useful in the treatment of life-threatening pulmonary embolism in patients with absolute contraindications to thrombolytic therapy. A larger cohort of patients is necessary to determine whether this treatment should be proposed as an alternative to the use of fibrinolytics in selected patients. PMID- 9355884 TI - Negative inotropic effect of basic fibroblast growth factor on adult rat cardiac myocyte. AB - BACKGROUND: Basic fibroblast growth factor (bFGF) is highly expressed in the myocardium in some cardiac disorders, such as ischemia-reperfusion and cardiac allograft rejection. However, whether bFGF has any effects on myocardial contraction is unknown. METHODS AND RESULTS: We examined the effects of bFGF on myocardial contractility using isolated adult rat cardiac myocyte preparations. bFGF exerted a direct negative inotropic effect that was concentration and time dependent. The pretreatment of myocytes with a neutralizing anti-bFGF antibody (100 ng/mL) abolished the negative inotropic effects of bFGF (100 ng/mL). Platelet-derived growth factor (12.5 ng/mL) and transforming growth factor-beta (1 ng/mL) did not exert such effects, which indicated that bFGF-induced negative inotropism was considered to be specific for this growth factor. bFGF decreased the peak intracellular Ca2+ transient by 46% during systole. The enhanced production of nitric oxide was unlikely to be responsible for the bFGF-induced negative inotropic effect. CONCLUSIONS: bFGF, primarily a potent growth promoter, produced acute negative inotropic effects in the adult cardiac myocyte that could have resulted from alterations in intracellular Ca2+ homeostasis. The negative inotropic effect of bFGF may contribute to myocardial dysfunction associated with ischemia-reperfusion injury and heart transplant rejection. PMID- 9355885 TI - Simplified electrophysiologically directed catheter ablation of recurrent common atrial flutter. AB - BACKGROUND: Despite verification of bidirectional conduction block after radiofrequency (RF) catheter ablation in the inferior vena cava (IVC)-tricuspid annulus (TA) isthmus, recurrence of common atrial flutter is relatively common. Although complete linear reablation is usually performed, we evaluated a simplified electrophysiological strategy selectively targeting recovered conducting isthmus tissue. METHODS AND RESULTS: Twenty-one patients (18 men and 3 women, age, 54+/-10 years) with a recurrence of typical atrial flutter 6+/-7 months after an apparently successful catheter ablation in the IVC-TA isthmus prospectively underwent electrophysiologically targeted reablation during flutter. Sites with narrow electrograms or fractionated electrograms interposed between adjacent sites with double potentials considered to represent gaps were ablated without movement of the catheter. Mapping showed that 18 of 21 patients had a single gap. Successful ablation required a single application in 14 patients and, in the group as a whole, a median of one application (mean, 2+/-2; range, 1 to 11) with resultant bidirectional block in 13 of 16. A single narrow electrogram (duration, 48+/-6 ms; amplitude, 0.1+/-0.05 mV) was noted at the successful site in 11, whereas a fractionated electrogram (97+/-32 ms, 0.05+/ 0.04 mV, P<.05) was noted in 9. There were four additional recurrences during a follow-up at 7+/-5 months; three were similarly ablated with a median of one pulse. CONCLUSIONS: Transmural ablation lesions in the isthmus can be recognized during flutter by double potentials separated by an isoelectric interval. Postablation recurrent flutter is usually due to a single discrete recovered gap; this is represented by a single or a fractionated potential spanning the isoelectric interval of adjacent double potentials, which can be selectively targeted to minimize repeat ablation. PMID- 9355886 TI - Vagal and sympathetic mechanisms in patients with orthostatic vasovagal syncope. AB - BACKGROUND: Autonomic and particularly sympathetic mechanisms play a central role in the pathophysiology of vasovagal syncope. We report direct measurements of muscle sympathetic nerve activity in patients with orthostatic vasovagal syncope. METHODS AND RESULTS: We studied 53 otherwise healthy patients with orthostatic syncope. We measured RR intervals and finger arterial pressures and in 15 patients, peroneal nerve muscle sympathetic activity before and during passive 60 degree head-up tilt, with low-dose intravenous isoproterenol if presyncope did not develop by 15 minutes. We measured baroreflex gain before tilt with regression of RR intervals or sympathetic bursts on systolic or diastolic pressures after sequential injections of nitroprusside and phenylephrine. Orthostatic vasovagal reactions occurred in 21 patients, including 7 microneurography patients. Presyncopal and nonsyncopal patients had similar baseline RR intervals, arterial pressure, and muscle sympathetic nerve activity. Vagal baroreflex responses were significantly impaired at arterial pressures below (but not above) baseline levels in presyncopal patients. Initial responses to tilt were comparable; however, during the final 200 seconds of tilt, presyncopal patients had lower RR intervals and diastolic pressures than nonsyncopal patients and gradual reduction of arterial pressure and sympathetic activity. Frank presyncope began abruptly with precipitous reduction of arterial pressure, disappearance of muscle sympathetic nerve activity, and RR interval lengthening. CONCLUSIONS: Patients with orthostatic vasovagal reactions have impaired vagal baroreflex responses to arterial pressure changes below resting levels but normal initial responses to upright tilt. Subtle vasovagal physiology begins before overt presyncope. The final trigger of human orthostatic vasovagal reactions appears to be the abrupt disappearance of muscle sympathetic nerve activity. PMID- 9355887 TI - Apolipoprotein(a) induces monocyte chemotactic activity in human vascular endothelial cells. AB - BACKGROUND: Elevated levels of lipoprotein(a) [Lp(a)] are associated with premature atherosclerosis; however, the mechanisms are not known. Recruitment of monocytes to the blood vessel wall is an early event in atherogenesis. METHODS AND RESULTS: This study has found that unoxidized Lp(a) induced human umbilical vein endothelial cells (HUVECs) to secrete monocyte chemotactic activity (MCA), whereas LDL under the same conditions did not. In the absence of HUVECs, Lp(a) had no direct MCA. Endotoxin was shown not to be responsible for the induction of MCA. Actinomycin D and cycloheximide inhibited the HUVEC response to Lp(a), indicating that protein and RNA synthesis were required. The apolipoprotein(a) [apo(a)] portion of Lp(a) was identified as the structural component of Lp(a) responsible for inducing MCA. Lp(a) and apo(a) also stimulated human coronary artery endothelial cells to produce MCA. Granulocyte-monocyte colony-stimulating factor (GM-CSF) antigen was not detected in the Lp(a)-conditioned medium, nor was monocyte chemoattractant protein-1 mRNA induced in HUVECs by Lp(a). CONCLUSIONS: These findings suggest that Lp(a) may be involved in the recruitment of monocytes to the vessel wall and provide a novel mechanism for the participation of Lp(a) in the atherogenic process. PMID- 9355888 TI - Fasting triglycerides, high-density lipoprotein, and risk of myocardial infarction. AB - BACKGROUND: Recent data suggest that triglyceride-rich lipoproteins may play a role in atherogenesis. However, whether triglycerides, as a marker for these lipoproteins, represent an independent risk factor for coronary heart disease remains unclear, despite extensive research. Several methodological issues have limited the interpretability of the existing data. METHODS AND RESULTS: We examined the interrelationships of fasting triglycerides, other lipid parameters, and nonlipid risk factors with risk of myocardial infarction among 340 cases and an equal number of age-, sex-, and community-matched control subjects. Cases were men or women of <76 years of age with no prior history of coronary disease who were discharged from one of six Boston area hospitals with the diagnosis of a confirmed myocardial infarction. In crude analyses, we observed a significant association of elevated fasting triglycerides with risk of myocardial infarction (relative risk [RR] in the highest compared with the lowest quartile=6.8; 95% confidence interval [CI]=3.8 to 12.1; P for trend <.001). Results were not materially altered after control for nonlipid coronary risk factors. As expected, the relationship was attenuated after adjustment for HDL but remained statistically significant (RR in the highest quartile=2.7; 95% confidence interval [CI]=1.4 to 5.5; P for trend=.016). Furthermore, the ratio of triglycerides to HDL was a strong predictor of myocardial infarction (RR in the highest compared with the lowest quartile=16.0; 95% CI=7.7 to 33.1; P for trend <.001). CONCLUSIONS: Our data indicate that fasting triglycerides, as a marker for triglyceride-rich lipoproteins, may provide valuable information about the atherogenic potential of the lipoprotein profile, particularly when considered in context of HDL levels. PMID- 9355889 TI - Effects of diet and sexual maturation on low-density lipoprotein cholesterol during puberty: the Dietary Intervention Study in Children (DISC). AB - BACKGROUND: The Dietary Intervention Study in Children (DISC) is a multicenter, randomized, controlled clinical trial designed to examine the efficacy and safety of a dietary intervention to reduce serum LDL cholesterol (LDL-C) in children with elevated LDL-C. METHODS AND RESULTS: The effects of dietary intake of fat and cholesterol and of sexual maturation and body mass index (BMI) on LDL-C were examined in a 3-year longitudinal study of 663 boys and girls (age 8 to 10 years at baseline) with elevated LDL-C levels. Multiple linear regression was used to predict LDL-C at 3 years. For boys, LDL-C decreased by 0.018 mmol/L for each 10 mg/4.2 MJ decrease in dietary cholesterol (P<.05). For girls, no single nutrient was significant in the model, but a treatment group effect was evident (P<.05). In both sexes, BMI at 3 years and LDL-C at baseline were significant and positive predictors of LDL-C levels. In boys, the average LDL-C level was 0.603 mmol/L lower at Tanner stage 4+ than at Tanner stage 1 (P<.01). In girls, the average LDL-C level was 0.274 mmol/L lower at Tanner stage 4+ than at Tanner stage 1 (P<.05). CONCLUSIONS: In pubertal children, sexual maturation, BMI, dietary intervention (in girls), and dietary cholesterol (in boys) were significant in determining LDL-C. Sexual maturation was the factor associated with the greatest difference in LDL-C. Clinicians screening for dyslipidemia or following dyslipidemic children should be aware of the powerful effects of pubertal change on measurements of lipoproteins. PMID- 9355891 TI - Hyperhomocyst(e)inemia is a risk factor for arterial endothelial dysfunction in humans. AB - BACKGROUND: Hyperhomocyst(e)inemia is associated with premature peripheral vascular, cerebrovascular, and coronary artery disease. Because homocysteine has been found to be damaging to endothelial cells in animal and cell culture studies, we evaluated the association between hyperhomocysteinemia and arterial endothelial dysfunction (a marker of early atherosclerosis) in asymptomatic adult subjects. METHODS AND RESULTS: Using high-resolution ultrasound, we measured endothelium-dependent flow-mediated dilation (EDD) and endothelium-independent nitroglycerin-induced dilation (GTN) of the brachial artery in 14 prospectively defined hyperhomocysteinemic (mean plasma homocysteine, 34.8+/-8.5 micromol/L), nonsmoking, healthy subjects aged 53+/-9 years and 14 control subjects with low plasma homocysteine levels (9.9+/-3.2 micromol/L). The two groups were well matched for age; sex; body mass index; blood pressure, blood cholesterol, folate, and vitamin B12 levels; and vessel diameter. EDD was significantly lower in hyperhomocysteinemic subjects (6.5+/-1.7%) than in subjects with low homocysteine levels (10.8+/-1.7%) (P<.001). GTN responses were similar in the two subject groups (P=.90). Multivariate analysis confirmed homocysteine level as the strongest predictor for impaired EDD, independent of age, sex, body mass index, or blood pressure, folate, vitamin B12, and cholesterol levels. CONCLUSIONS: Hyperhomocysteinemia is an independent risk factor for arterial endothelial dysfunction in healthy middle-aged adults. PMID- 9355890 TI - Attenuated progression of coronary artery disease after 6 years of multifactorial risk intervention: role of physical exercise. AB - BACKGROUND: It was the aim of this study to assess the long-term effects of physical exercise and low-fat diet on the progression of coronary artery disease. At the beginning of the study, 113 male patients with coronary artery disease were randomized to an intervention group (n=56) or a control group (n=57); 90 patients (80%) could be reevaluated after 6 years. METHODS AND RESULTS: Patients in the intervention group (n=40) showed a reduction in total serum cholesterol (6.03+/-1.03 versus 5.67+/-1.01 mmol/L; P<.03) and triglyceride levels (1.94+/ 0.8 versus 1.6+/-0.89 mmol/L; P<.005) and maintained their initial body mass index (26+/-2 versus 27+/-2 kg/m2; P=NS), but results were not statistically different from the control group (n=50) (total serum cholesterol, 6.05+/-1.02 versus 5.79+/-0.88 mmol/L; triglycerides, 2.25+/-1.28 versus 1.85+/-0.96 mmol/L [both P=NS]; body mass index, 26+/-2 versus 28+/-3 kg/m2 [P<.0001]). In the intervention group, there was a significant 28% increase in physical work capacity (166+/-59 versus 212+/-89 W; P<.001), whereas values remained essentially unchanged in the control group (165+/-51 versus 170+/-60 W; P=NS; between groups, P<.05). In the intervention group, coronary stenoses progressed at a significantly slower rate than in the control group (P<.0001). Energy expenditure during exercise was assessed in a subgroup; patients with regression of coronary stenoses spent an average of 1784+/-384 kcal/wk (approximately 4 hours of moderate aerobic exercise per week). Multivariate regression analysis identified only physical work capacity as independently contributing to angiographic changes. CONCLUSIONS: After 6 years of multifactorial risk intervention, there is significant and persistent improvement in lipoprotein levels and physical work capacity, which results in a significant retardation of disease progression. These beneficial effects appear to be largely due to chronic physical exercise. PMID- 9355892 TI - Randomized, double-blind, placebo-controlled study of supplemental vitamin E on attenuation of the development of nitrate tolerance. AB - BACKGROUND: The attenuation of intracellular production of cGMP has been known to be a mechanism of nitrate tolerance. A recent in vitro study showed an increase in superoxide levels and a reduced activation of guanylate cyclase in tolerant vessels. We investigated the preventive effect of an antioxidant, vitamin E, on the development of nitrate tolerance. METHODS AND RESULTS: In this double-blind, placebo-controlled study, 24 normal volunteers and 24 patients with ischemic heart disease (IHD patients) were randomized to receive either vitamin E (200 mg TID vitamin E group) or placebo (placebo group). Vasodilator response to nitroglycerin was assessed with forearm plethysmography by measurement of the change in the forearm blood flow before and 5 minutes after sublingual administration of 0.3 mg nitroglycerin, and at the same time, blood samples were taken from veins to measure the platelet cGMP level. Measurements of the forearm blood flow and blood sampling were obtained serially at baseline (day 0), 3 days after vitamin E or placebo alone was taken (day 3), and 3 days after application of a 10-mg/24-h nitroglycerin tape concomitantly with oral vitamin E or placebo (day 6). The responses of forearm blood flow (%FBF) and cGMP (%cGMP) after sublingual nitroglycerin on day 0 (%FBF: normal volunteers, 32+/-12 versus 31+/ 11; IHD patients, 35+/-15 versus 34+/-15; %cGMP: normal volunteers, 38+/-10 versus 35+/-11; IHD patients, 37+/-11 versus 38+/-12, vitamin E group versus placebo group) and day 3 (%FBF: normal volunteers, 33+/-9 versus 32+/-12; IHD patients, 35+/-12 versus 33+/-13; %cGMP: normal volunteers, 38+/-10 versus 37+/ 11; IHD patients, 36+/-14 versus 37+/-10, vitamin E group versus placebo group) were not different between the two groups. On day 6, %FBF and %cGMP in the placebo group were significantly lower compared with day 0, and there were significant differences in them between the two groups (%FBF: normal volunteers, 30+/-12 versus 17+/-9, P<.01; IHD patients, 28+/-14 versus 17+/-8, P<.01; %cGMP: normal volunteers, 35+/-11 versus 8+/-5, P<.01; IHD patients, 38+/-10 versus 12+/ 4, P<.01, vitamin E group versus placebo group). CONCLUSIONS: These results indicate that the combination therapy with vitamin E is potentially a useful method to prevent the development of nitrate tolerance. PMID- 9355893 TI - Relationship between diabetes mellitus and long-term survival after coronary bypass and angioplasty. AB - BACKGROUND: Recent subgroup analyses of randomized trials have suggested that percutaneous intervention in diabetic patients with multivessel disease results in higher mortality than coronary artery bypass graft surgery (CABG). We studied the relationship between diabetes and survival after revascularization in a large prospective cohort of patients with multivessel coronary artery disease. METHODS AND RESULTS: By analyzing data for 3220 patients (24% diabetic) with symptomatic two- or three-vessel coronary disease who were undergoing percutaneous transluminal coronary angioplasty (PTCA) or CABG at Duke University Medical Center between 1984 and 1990, we found that at 5 years, unadjusted survival in the group of patients undergoing CABG was 74% in diabetics and 86% in nondiabetics. Similarly, 5-year survival among PTCA patients was 76% in diabetics and 88% in patients without diabetes. After adjustment for baseline characteristics, diabetic patients receiving either PTCA or CABG had significantly poorer survival than nondiabetics (chi2=43.56, P<.0001). Unlike previous studies, however, there was no significant differential effect of diabetes on outcome between patients treated with PTCA and those treated with CABG (chi2=0.01, P=.91). CONCLUSIONS: Although diabetes was associated with a worse long-term outcome after both PTCA and CABG in patients with multivessel coronary artery disease, the effect of diabetes on prognosis was similar in both treatment groups. Thus, our findings support the concept that the choice of initial revascularization strategy should not be based exclusively on a history of diabetes but rather should rely on other factors, such as angiographic suitability and clinical status. PMID- 9355894 TI - Efficacy of mibefradil compared with amlodipine in suppressing exercise-induced and daily silent ischemia: results of a multicenter, placebo-controlled trial. AB - BACKGROUND: Mibefradil is a new benzimidazolyl-substituted tetraline-derivative calcium antagonist. Its vasodilatory activity combined with an ability to lower heart rate without negative inotropic effects as well as its long duration of action make it a promising anti-ischemic agent. METHODS AND RESULTS: Three hundred nine patients with coronary artery disease, stable angina pectoris, and positive exercise tests were randomized to receive mibefradil (50, 100, or 150 mg), amlodipine (10 mg), or placebo. The anti-ischemic effects of mibefradil on exercise test and silent ischemia parameters were assessed. At doses of 100 and 150 mg, mibefradil increased exercise duration (by 55.5 and 51.0 seconds, respectively; P<.001 for both), increased time to onset of angina (by 98.3 and 82.7 seconds, respectively; P<.001), and increased time to 1-mm ST depression (by 81.7 and 94.3 seconds, respectively; P<.001). By comparison, a 10 mg/d dose of amlodipine significantly improved only time to onset of angina (treatment effect: 38.5 seconds, P=.036). Mibefradil 100 mg and 150 mg decreased the number of episodes of silent ischemia (treatment effects: -3.1 and -3.6, respectively; P<.001) and the duration of silent ischemia (treatment effects: -9.2 minutes, P=.048, and -14.6 minutes, P=.002, respectively). The decrease in the number of episodes of silent ischemia was also statistically significant in the group receiving 10 mg of amlodipine (-1.5; P=.036). CONCLUSIONS: Once-daily doses of 100 and 150 mg mibefradil were effective in improving exercise tolerance and reducing ischemic episodes during ambulatory monitoring in patients with coronary artery disease. PMID- 9355895 TI - Collagen scar formation after acute myocardial infarction: relationships to infarct size, left ventricular function, and coronary artery patency. AB - BACKGROUND: Left ventricular function after acute myocardial infarction (AMI) is determined by the expansion of the infarct zone and remodeling of the noninfarcted myocardium. An occluded infarct-related artery (IRA) is an independent risk factor for remodeling. METHODS AND RESULTS: Changes in myocardial collagen metabolism were evaluated in 36 patients with suspected AMI. The plasma creatine kinase MB fraction and myoglobin release curves were analyzed for assessment of early reperfusion and infarct size. Collagen scar formation was evaluated by measurement of serum concentrations of the aminoterminal propeptide of type III procollagen (PIIINP), the aminoterminal propeptide of type I procollagen (intact PINP), and the carboxyterminal propeptide of type I procollagen (PICP). Plasma renin activity and urine excretion of cortisol and aldosterone were also measured. Coronary angiography and left ventricular cineangiography were performed during early hospitalization. The serum concentration of PIIINP increased from 3.50+/-0.20 to a maximum of 5.08+/-0.36 microg/L (n=32) in the patients with AMI, whereas the concentrations of intact PINP and PICP tended to decrease. The area under the curve (AUC) of PIIINP during the first 10 postinfarction days was larger in patients with severe heart failure or ejection fractions < or = 40% than in those with no heart failure or with an ejection fraction > 40% (P<.05 and P<.01, respectively), and it was also larger in the patients with TIMI grade 0 to 2 flows than in those with TIMI 3 flows (P<.05), despite similar enzymatically determined infarct sizes. No significant correlations between PIIINP and neurohumoral parameters were observed. The AUC of PIIINP and the change in PIIINP during the first 4 days were significantly correlated with indices of cardiac function. CONCLUSIONS: Collagen scar formation after AMI can be quantified by measurement of serum PIIINP concentrations. Scar formation is more prominent in large infarctions causing left ventricular dysfunction and in patients with occluded IRAs. PMID- 9355896 TI - Thermolabile methylenetetrahydrofolate reductase in coronary artery disease. AB - BACKGROUND: Hyperhomocysteinemia, an independent and graded risk factor for coronary artery disease (CAD), may result from both environmental and hereditary factors. Methylenetetrahydrofolate reductase (MTHFR) catalyzes the conversion of methylenetetrahydrofolate to methyltetrahydrofolate, the methyl donor in the remethylation of homocysteine to methionine. A 677C-->T mutation in the MTHFR gene has been associated with elevated homocysteine concentrations in homozygous (+/+) individuals. METHODS AND RESULTS: We assessed the frequency of this common mutation in 735 CAD patients from the Regression Growth Evaluation Statin Study (REGRESS), a lipid-lowering coronary-regression trial, and in 1250 population based control subjects. Furthermore, the association between the mutation and serum homocysteine concentrations was studied. The frequency of the homozygous (+/+) mutation was 9.5% among patients versus 8.5% among control subjects, resulting in an odds ratio of 1.21 (95% confidence interval [CI], 0.87 to 1.68), relative to the (-/-) genotype. Homocysteine concentrations were significantly elevated in both (+/+) and (+/-) individuals compared with (-/-) individuals (median homocysteine levels, 15.4, 13.4, and 12.6 micromol/L, for (+/+), (+/-), and (-/-) individuals, respectively). For a summary estimation of the risk of the (+/+) genotype for CAD, we performed a meta-analysis on 8 different case-control studies on thermolabile MTHFR in CAD. In the meta-analysis, the homozygous (+/+) genotype was present in 299 of 2476 patients (12.1%) and in 257 (10.4%) of 2481 control subjects, resulting in a significant odds ratio of 1.22 (95% CI, 1.01 to 1.47) relative to the (-/-) genotype. CONCLUSIONS: Both the homozygous (+/+) and heterozygous (+/-) genotype result in elevated homocysteine concentrations. From our meta-analysis, we conclude that the homozygous (+/+) genotype is a modest but significant risk factor for CAD. PMID- 9355897 TI - Applicability of cardiac troponin T and I for early risk stratification in unstable coronary artery disease. TRIM Study Group. Thrombin Inhibition in Myocardial ischemia. AB - BACKGROUND: Studies have demonstrated that troponin T is a strong independent indicator of a poor prognosis in patients with unstable coronary artery disease. Up to the present, no study has compared the prognostic value of troponin T with that of troponin I in the same cohort of patients. METHODS AND RESULTS: Patients (n=516) suspected of having unstable coronary artery disease were investigated. Follow-up was done after 30 days, and the occurrences of cardiac death, acute myocardial infarction, refractory angina pectoris, and recurrent angina pectoris were registered. Elevated levels of troponin T (> or = 0.10 microg/L) were associated with an increased risk of cardiac death at 30 days compared with patients with normal levels, 3.2% versus 0.4% (P=.014). Troponin I values above the chosen cutoff (2.0 microg/L) were similarly found to be an indicator of increased risk of cardiac death, 3.2% versus 0.7% (P=.026). With regard to the composite end point of cardiac death/acute myocardial infarction, the troponins were strong independent indicators of adverse outcome. CONCLUSIONS: In patients suspected of having unstable coronary artery disease, both troponin T and troponin I provide independent prognostic information with regard to cardiac death and acute myocardial infarction. PMID- 9355898 TI - Augmented peripheral chemosensitivity as a potential input to baroreflex impairment and autonomic imbalance in chronic heart failure. AB - BACKGROUND: The precise mechanisms responsible for the sympathetic overactivity and blunted baroreflex control in chronic heart failure (CHF) remain obscure. Augmented peripheral chemosensitivity has recently been demonstrated in CHF. We evaluated the relation between peripheral chemoreflex sensitivity and autonomic activity in patients with CHF. METHODS AND RESULTS: We studied in 26 stable patients with CHF the peripheral chemosensitivity (ventilatory response to hypoxia using transient inhalations of pure nitrogen), autonomic balance (spectral analysis of heart rate variability [HRV]), and baroreflex sensitivity (bolus phenylephrine method and alpha index). To determine whether transient inactivation of peripheral chemoreceptors might influence autonomic balance, 12 patients underwent a second study during which they breathed 100% O2. Peripheral chemosensitivity correlated inversely with HRV power within the low-frequency band (0.04 to 0.15 Hz) (r=-.52, P=.006) and inversely with baroreflex sensitivity (r=-.60, P=.005). When the patients were divided into two groups according to the chemosensitivity of age-matched normal controls (above and below mean+2 SDs of chemosensitivity of control subjects), those above the normal range revealed more impaired autonomic balance, ie, lower baroreflex sensitivity (1.4 +/- 1.3 versus 5.0 +/- 1.5 ms/mm Hg, P<.0001) and depressed values of low-frequency power (2.5 +/- 1.8 versus 4.1 +/- 0.8 ln ms2, P<.005) compared with those with normal chemosensitivity. Transient hyperoxia did not alter heart rate or systolic pressure but resulted in an increase in HRV and an improvement in baroreflex sensitivity. CONCLUSIONS: A link between increased peripheral chemosensitivity and impaired autonomic control, including baroreflex inhibition, is demonstrated. The clinical importance of this phenomenon warrants further investigation. PMID- 9355899 TI - Arrhythmogenic marker for the sudden unexplained death syndrome in Thai men. AB - BACKGROUND: Between 1981 and 1988, the Centers for Disease Control and Prevention reported a very high incidence of sudden death among young male Southeast Asians who died unexpectedly during sleep. The pattern of death has long been prevalent in Southeast Asia. We carried out a study to identify the clinical markers for patients at high risk of developing sudden unexplained death syndrome (SUDS) and long-term outcomes. METHODS AND RESULTS: We studied 27 Thai men (mean age, 39.7+/ 11 years) referred because they had cardiac arrest due to ventricular fibrillation, usually occurring at night while asleep (n=17), or were suspected to have had symptoms similar to the clinical presentation of SUDS (n=10). We performed cardiac testing, including EPS and cardiac catheterization. The patients were then followed at approximately 3-month intervals; our primary end points were death, ventricular fibrillation, or cardiac arrest. A distinct ECG abnormality divided our patients who had no structural heart disease (except 3 patients with mild left ventricular hypertrophy) into two groups: group 1 (n=16) patients had right bundle-branch block and ST-segment elevation in V1 through V3, and group 2 (n=11) had a normal ECG. Group 1 patients had well-defined electrophysiological abnormalities: group 1 had an abnormally prolonged His Purkinje conduction time (HV interval, 63+/-11 versus 49+/-6 ms; P=.007). Group 1 had a higher incidence of inducible ventricular fibrillation (93% for group 1 versus 11% for group 2; P=.0002) and a positive signal-averaged ECG (92% for group 1 versus 11% for group 2; P=.002), which was associated with a higher incidence of ventricular fibrillation or death (P=.047). The life-table analysis showed that the group 1 patients had a much greater risk of dying suddenly (P=.05). CONCLUSIONS: Right bundle-branch block and precordial injury pattern in V1 through V3 is common in SUDS patients and represents an arrhythmogenic marker that identifies patients who face an inordinate risk of ventricular fibrillation or sudden death. PMID- 9355900 TI - Characterization of low right atrial isthmus as the slow conduction zone and pharmacological target in typical atrial flutter. AB - BACKGROUND: Previous electrophysiological studies in patients with typical atrial flutter suggested that the slow conduction zone might be located in the low right atrial isthmus, which is a path formed by orifice of inferior vena cava, eustachian valve/ridge, coronary sinus ostium, and tricuspid annulus. The conduction characteristics during atrial pacing and responses to antiarrhythmic drugs of this anatomic isthmus were unknown. METHODS AND RESULTS: Forty-four patients, 20 patients with paroxysmal supraventricular tachycardia (group 1) and 24 patients with clinically documented paroxysmal typical atrial flutter (group 2), were studied. A 20-pole halo catheter was situated around the tricuspid annulus. Incremental pacing from the low right atrium and coronary sinus ostium was performed to measure the conduction time and velocity along the isthmus and lateral wall in the baseline state and after intravenous infusion of procainamide or sotalol. In both groups, conduction velocity in the isthmus during incremental pacing was significantly lower than that in the lateral wall before and after infusion of antiarrhythmic drugs. Furthermore, gradual conduction delay with unidirectional block in the isthmus was relevant to initiation of typical atrial flutter. Compared with group 1, group 2 had a lower conduction velocity in the isthmus and shorter right atrial refractory period. Procainamide significantly decreased the conduction velocity, but sotalol did not change it. In contrast, sotalol significantly prolonged the atrial refractory period with a higher extent than procainamide. After infusion of procainamide, the increase of conduction time in the isthmus accounted for 52+/-19% of the increase in flutter cycle length, and 5 of 12 patients (42%) had spontaneous termination of typical flutter. After infusion of sotalol, typical flutter was induced in only 6 of 12 patients (50%) without significant prolongation of flutter cycle length. CONCLUSIONS: The low right atrial isthmus with rate-dependent slow conduction properties is critical to initiation of typical human atrial flutter. It may be the potentially pharmacological target of antiarrhythmic drugs in the future. PMID- 9355901 TI - Prediction of transition to chronic atrial fibrillation in patients with paroxysmal atrial fibrillation by signal-averaged electrocardiography: a prospective study. AB - BACKGROUND: It is well known that paroxysmal atrial fibrillation (PAF) often precedes the establishment of chronic atrial fibrillation (CAF). However, there have been no definite methods to predict the transition from PAF to CAF. The purpose of this report was to determine prospectively whether P-wave-triggered signal-averaged ECG (P-SAE) is useful for the prediction of the transition to CAF in patients with PAF. METHODS AND RESULTS: One hundred twenty-two consecutive patients with PAF were prospectively followed after P-SAE, echocardiography, and 24-hour Holter monitoring at study entry. The duration (Ad) and root-mean-square voltage for the last 30 ms (LP30) of the filtered P wave were measured in P-SAE. The abnormality of P-SAE for the prediction of transition to CAF was defined as Ad > or = 145 ms and LP30 < 3.0 microV. Twenty-three (19%; group 1) of the patients had the abnormality of P-SAE, whereas the others (group 2) did not. During the follow-up period (mean, 26+/-12 months), 10 patients (43%) in group 1 acquired CAF, whereas the transition to CAF was observed in only 4 patients (4%) in group 2. Kaplan-Meier analysis revealed that the transition to CAF was significantly observed more often in group 1 than in group 2 (log-rank test, P<.0001). The Cox proportional hazards regression model identified that the variables most significantly associated with the transition to CAF were Ad (chi2=8.6, P=.003) and LP30 (chi2=5.1, P=.02), although significant differences in the left atrial dimension (40.8+/-5.3 versus 37.3+/-5.5 mm, P<.01) and the number of atrial premature contractions (3641+/-4524 versus 1489+/-2895 beats/d, P<.05) were observed between groups 1 and 2. CONCLUSIONS: These results indicate that P-SAE could be useful to identify patients at risk for the transition from PAF to CAF. PMID- 9355902 TI - Assessment of atrioventricular junction ablation and DDDR mode-switching pacemaker versus pharmacological treatment in patients with severely symptomatic paroxysmal atrial fibrillation: a randomized controlled study. AB - BACKGROUND: The purpose of the study was to evaluate the effect of AV junction ablation and pacemaker implantation on quality of life and specific symptoms in patients with paroxysmal atrial fibrillation (AF) not controlled by drugs. METHODS AND RESULTS: We performed a multicenter, randomized, 6-month evaluation of the clinical effects of AV junction ablation and DDDR mode-switching pacemaker (Abl+Pm) versus pharmacological treatment in 43 patients with intolerable, recurrent paroxysmal AF of three or more episodes in the previous 6 months not controlled with three or more antiarrhythmic drugs. Before completion of the study, 3 patients in the drug group withdrew because of the severity of their symptoms and 1 patient assigned to the Abl+Pm group in whom the ablation procedure failed. At the end of the 6 months, the 21 patients of the Abl+Pm group who completed the study showed, in comparison with the 18 of the drug group, lower scores in the Living with Heart Failure Questionnaire (-51%, P=.0006), palpitations (-71%, P=.0000), effort dyspnea (-36%, P=.04), exercise intolerance score (-46%, P=.001), and easy fatigue (-51%, P=.02). The scores for rest dyspnea, chest discomfort, and NYHA functional classification were also lower ( 56%, -50%, and -17%, respectively) in the Abl+Pm group, although not significantly. At the end of the study, palpitations were no longer present in 81% of the Abl+Pm group and in 11% of the drug group (P=.0000). AF was documented in 31 of 122 visits (25%) in the Abl+Pm group and in 9 of 107 examinations (8%) in the drug group (P=.0005); chronic AF developed in 5 (24%) and 0 (0%) in the two groups, respectively (P=.04). CONCLUSIONS: In patients with paroxysmal AF not controlled by pharmacological therapy, Abl+Pm treatment is highly effective and superior to drug therapy in controlling symptoms and improving quality of life. The discontinuation of drug therapy exposes patients to further recurrences of paroxysmal AF and the risk of developing permanent AF. PMID- 9355903 TI - Treatment of atrial fibrillation and paroxysmal supraventricular tachycardia with bidisomide. The Atrial Fibrillation Investigation with Bidisomide (AFIB) Investigators. AB - BACKGROUND: Atrial fibrillation and paroxysmal supraventricular tachycardia are common disorders of the heart rhythm for which antiarrhythmic drug therapy is commonly prescribed. The Atrial Fibrillation Investigation with Bidisomide (AFIB) study was a randomized, placebo-controlled clinical trial designed to accomplish three goals in a single protocol: (1) to determine the efficacy of the antiarrhythmic drug bidisomide in the treatment of these two arrhythmias; (2) to establish the appropriate dose range for bidisomide; and (3) to detect an adverse mortality effect of bidisomide if one were present in patients with atrial fibrillation. METHODS AND RESULTS: In this clinical trial, 1227 patients with atrial fibrillation and 187 with paroxysmal supraventricular tachycardia were randomly assigned to bidisomide (200, 400, or 600 mg BID) or placebo; patient groups with each arrhythmia were analyzed separately. Symptomatic recurrences of atrial fibrillation and paroxysmal supraventricular tachycardia were documented with the use of transtelephonic ECG monitoring. The time to the first symptomatic arrhythmia recurrence was measured in each patient and compared among treatment groups. Among the atrial fibrillation patients, there was no significant difference in the time to first symptomatic recurrence between the placebo group and any of the three bidisomide treatment groups; the hazard ratios (placebo:treatment) were 1.19, 1.03, and 1.14 for bidisomide 200, 400, and 600 mg BID, respectively. Among paroxysmal supraventricular tachycardia patients, there was a similar lack of a significant treatment effect; the hazard ratios were 1.30, 1.93, and 1.59 for bidisomide 200, 400, and 600 mg BID, respectively. In the primary safety analysis of mortality, 3 of 493 patients taking placebo died, compared with 9 of 488 patients taking one of the two higher doses of bidisomide (P>.10). CONCLUSIONS: Bidisomide in the doses tested did not have a clinically important antiarrhythmic effect. The AFIB study provided a novel clinical trial design to test antiarrhythmic drugs for both safety and efficacy. PMID- 9355904 TI - Significance of morphological abnormalities detected by MRI in patients undergoing successful ablation of right ventricular outflow tract tachycardia. AB - BACKGROUND: MRI can demonstrate subtle morphological changes of the right ventricle in patients with idiopathic right ventricular outflow tract tachycardia (RVOT). The present study examines the incidence and significance of right ventricular (RV) abnormalities detected by MRI with respect to the site of successful radiofrequency catheter ablation of the clinical tachycardia. METHODS AND RESULTS: The study population comprised 20 patients (mean age, 40+/-12 years) undergoing elimination of recurrent RVOT by radiofrequency catheter ablation. MRI studies were performed before ablation to assess RV volumes and function, as well as structural abnormalities of the RV myocardium. Ten healthy age- and sex matched subjects served as control subjects. The successful ablation sites, as documented by radiographs of the catheter position, were compared with MRI findings. Patients with RVOT showed no difference in respect to RV volumes and ejection fractions compared with control subjects. Whereas RV abnormalities were limited to prominent fatty deposits of the right atrioventricular groove extending into the inlet portion of the RV wall in 2 of 10 control subjects, MRI studies demonstrated morphological changes of the RV free wall in 13 (65%) of 20 patients with RVOT, including presence of fatty tissue (n=5), wall thinning (n=9), and dyskinetic wall segments (n=4). Eight of these patients had additional fat deposits, thinning, or a saccular aneurysm in the RV outflow tract, corresponding with the ablation site in 6 patients. CONCLUSIONS: In RVOT, structural abnormalities of the right ventricle can be detected in a substantial number of patients despite normal RV volumes and global function. MRI abnormalities within the RV outflow tract are significantly associated with the origin of tachycardia. PMID- 9355906 TI - Protective effect of chronic garlic intake on elastic properties of aorta in the elderly. AB - BACKGROUND: Epidemiological studies have suggested that garlic may have protective effects against cardiovascular diseases. We undertook this cross sectional observational study to test the hypothesis that regular garlic intake would delay the stiffening of the aorta relating to aging. METHODS AND RESULTS: We studied healthy adults (n=101; age, 50 to 80 years) who were taking > or = 300 mg/d of standardized garlic powder for > or = 2 years and 101 age- and sex matched control subjects. Pulse wave velocity (PWV) and pressure-standardized elastic vascular resistance (EVR) were used to measure the elastic properties of the aorta. Blood pressures, heart rate, and plasma lipid levels were similar in the two groups. PWV (8.3+/-1.46 versus 9.8+/-2.45 m/s; P<.0001) and EVR (0.63+/ 0.21 versus 0.9+/-0.44 m2 x s(-2) x mm Hg(-1); P<.0001) were lower in the garlic group than in the control group. PWV showed significant positive correlation with age (garlic group, r=.44; control group, r=.52) and systolic blood pressure (SBP) (garlic group, r=.48; control group, r=.54). With any degree of increase in age or SBP, PWV increased less in the garlic group than in the control group (P<.0001). ANCOVA and multiple regression analyses demonstrated that age and SBP were the most important determinants of PWV and that the effect of garlic on PWV was independent of confounding factors. CONCLUSIONS: Chronic garlic powder intake attenuated age-related increases in aortic stiffness. These data strongly support the hypothesis that garlic intake had a protective effect on the elastic properties of the aorta related to aging in humans. PMID- 9355905 TI - Predictive value of cardiac troponin T in pediatric patients at risk for myocardial injury. AB - BACKGROUND: Biochemical markers have not been routinely used in children at risk for myocardial damage. Yet, because of somatic growth and the duration of survival, a low level of myocardial damage may ultimately be of more consequence in children than in adults. METHODS AND RESULTS: We investigated the utility of cardiac troponin T (cTnT) blood levels (CARDIAC T ELISA Troponin T, Boehringer Mannheim Corp) in 51 consecutively sampled patients from 1 day to 34 years of age (median=5.7 years) undergoing cardiovascular (n=19) or noncardiovascular (n=17) surgery or who received doxorubicin for acute lymphoblastic leukemia (ALL) (n=15). Minimum detectable cTnT elevations were 0.03 ng/mL. cTnT was measurable in children of all ages with myocyte damage. In patients who underwent cardiovascular surgery, a correlation was noted between a score of increasing surgical severity and the mean level of postoperative cTnT (r=.79, P<.0001). Postoperative cTnT levels were elevated in children who completed cardiovascular surgery with an open chest compared with those with a closed chest (P=.0083). In addition, cTnT levels before cardiovascular surgery predicted postoperative survival (P=.007). cTnT elevations were observed after initial doxorubicin therapy for ALL. The magnitude of elevation predicted left ventricular dilatation (r=.80 when variables were treated as continuous, P=.003) and wall thinning (r=.61, P=.044) 9 months later. CONCLUSIONS: Elevations of blood cTnT in children relate to the severity of myocardial damage and predict subsequent subclinical and clinical cardiac morbidity and mortality. PMID- 9355907 TI - Exogenous and endogenous adenosine inhibits fetal calf serum-induced growth of rat cardiac fibroblasts: role of A2B receptors. AB - BACKGROUND: Because proliferation of cardiac fibroblasts participates in cardiac hypertrophy/remodeling associated with hypertension and myocardial infarction, it is important to elucidate factors regulating cardiac fibroblast proliferation. Adenosine, a nucleoside abundantly produced by cardiac cells, is antimitogenic vis-a-vis vascular smooth muscle cells; however, the effect of adenosine on cardiac fibroblast proliferation is unknown. The objective of this study was to characterize the effects of exogenous and endogenous (cardiac fibroblast-derived) adenosine on cardiac fibroblast proliferation. METHODS AND RESULTS: Growth arrested cardiac fibroblasts were stimulated with 2.5% FCS in the presence and absence of adenosine, 2-chloroadenosine (stable adenosine analogue), or modulators of adenosine levels, including (1) erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA; adenosine deaminase inhibitor); (2) dipyridamole (adenosine transport blocker); and (3) iodotubericidin (adenosine kinase inhibitor). All of these agents inhibited, in a concentration-dependent manner, FCS-induced cardiac fibroblast proliferation as assessed by DNA synthesis ([3H]thymidine incorporation) and cell counting. EHNA, dipyridamole, and iodotubericidin increased extracellular levels of adenosine by 2.3- to 5.6-fold when added separately to cardiac fibroblasts, and EHNA+iodotubericidin or EHNA+iodotubericidin+dipyridamole increased extracellular adenosine levels by >690-fold. Both KF17837 (selective A2 antagonist) and DPSPX (nonselective A2 antagonist) but not DPCPX (selective A1 antagonist) blocked the antimitogenic effects of 2-chloroadenosine, EHNA, and dipyridamole on DNA synthesis, suggesting the involvement of A2A and/or A2B but excluding the participation of A1 receptors. The lack of effect of CGS21680 (selective A2A agonist) excluded involvement of A2A receptors and suggested a major role for A2B receptors. This conclusion was confirmed by the rank order potencies of four adenosine analogues. CONCLUSIONS: Cardiac fibroblasts synthesize adenosine, and exogenous and cardiac fibroblast-derived adenosine inhibits cardiac fibroblast proliferation via activation of A2B receptors. Cardiac fibroblast-derived adenosine may regulate cardiac hypertrophy and/or remodeling by modulating cardiac fibroblast proliferation. PMID- 9355908 TI - Hypercholesterolemia attenuates angiogenesis but does not preclude augmentation by angiogenic cytokines. AB - BACKGROUND: The impact of hyperlipidemia on collateral vessel development in vivo remains enigmatic. We sought to determine the anatomic extent and functional capacity of the collateral bed that develops in response to limb ischemia in a well characterized animal model of spontaneous hypercholesterolemia, the Watanabe heritable hyperlipidemic (WHHL) rabbit. We further characterized the impact of exogenous angiogenic cytokine administration on collateral vessel development and function in the same animal model. METHODS AND RESULTS: Weight-matched 6-month old male homozygous WHHL (n=9) and normal New Zealand White (NZW) (n=9) rabbits underwent surgical resection of one femoral artery. Ten days later, the ischemic hindlimb was evaluated for collateral vessel formation, blood flow, and tissue damage. Collateral vasculature was less extensive among WHHL than NZW, as indicated by a significant reduction in angiographic score (0.19+/-0.02 versus 0.35+/-0.03, P<.001) and capillary density (46.4+/-4.1 versus 78.9+/-4.6/mm2, P<.0002). This was associated with a reduction in calf blood pressure index (9.5+/-3.5% versus 32.8+/-2.8%, P<.0001), arterial blood flow (7.5+/-0.6 versus 13.6+/-0.7 mL/min, P<.0001), and muscle perfusion index (40.1+/-3.2% versus 65.9+/-2.0%, P<.0001) and an increase in muscle necrosis (48.16+/-5.41% versus 25.90+/-3.83% negative 2,3,5-triphenyltetrazolium chloride staining, P<.004). Treatment of WHHL rabbits (n=9) with recombinant human vascular endothelial growth factor produced a statistically significant improvement in all functional as well as anatomic indices of collateral development. CONCLUSIONS: Collateral vessel development associated with hindlimb ischemia in vivo is severely attenuated in an animal model of spontaneous hypercholesterolemia but nevertheless may be augmented by administration of angiogenic cytokines. PMID- 9355910 TI - Physical training alters the pathogenesis of pacing-induced heart failure through endothelium-mediated mechanisms in awake dogs. AB - BACKGROUND: Beneficial effects of exercise training on cardiovascular function in chronic heart failure (CHF) have been suggested previously, but the underlying mechanisms are unknown. We tested whether daily exercise training improves systemic hemodynamics and preserves endothelium-mediated vasodilator function during development of heart failure. METHODS AND RESULTS: Fifteen dogs were surgically instrumented for hemodynamic measurements. One group of dogs underwent 4 weeks of cardiac pacing (210 bpm for 3 weeks and 240 bpm during week 4), and another group underwent pacing plus daily exercise training (4.4+/-0.3 km/h, 2 h/d). Pacing-alone dogs developed CHF characterized by typical hemodynamic abnormalities, blunted endothelium-mediated vasodilator function in coronary and femoral circulations, and decreased gene expression of endothelial constitutive nitric oxide synthase (ECNOS, normalized to GAPDH expression; normal, 1.15+/-0.31 versus CHF, 0.29+/-0.08, P<.05). Exercise training preserved normal hemodynamics at rest, endothelium-mediated vasodilator function, and gene expression of ECNOS (0.72+/-0.16 versus normal, P=NS). Inhibition of NO synthesis (nitro-L-arginine) in exercise-trained dogs abolished the preserved endothelium-mediated vasodilation of epicardial coronary arteries and elevated left ventricular end diastolic pressure (7.7+/-0.3 to 19+/-3.4 mm Hg, P<.05), suggesting that the preservation of resting hemodynamics was in large part due to preserved endothelial function concealing the underlying CHF state. CONCLUSIONS: Long-term exercise training altered the natural history of heart failure due to rapid cardiac pacing. One of the underlying mechanisms is through the preservation of endothelial vasodilator function. PMID- 9355909 TI - Inotropic effects of glyceryl trinitrate and spontaneous NO donors in the dog heart. AB - BACKGROUND: In vitro, NO has a biphasic effect on myocardial inotropy. To determine the inotropic effect of NO in vivo, we investigated the activity of glyceryl trinitrate (GTN) and the NO donors S-nitroso-N-acetyl-D,L-penicillamine (SNAP) and sodium-(2)-1-(N,N-diethyl-amino)-diazen-1-ium-1,2-diolat+ ++ (DEA/NO) in dogs. METHODS AND RESULTS: Eight anesthetized open-chest dogs were instrumented for measurement of left ventricular and aortic pressures (tip manometers) and coronary flow (ultrasonic flow probes). Regional myocardial function was assessed by sonomicrometry as systolic wall thickening (sWT), mean systolic thickening velocity (Vs), and regional myocardial stroke work index (RSW). GTN, SNAP, and DEA/NO were infused into the left anterior descending coronary artery (LAD) to achieve defined coronary plasma concentrations of GTN, SNAP (both 10 to 100 micromol/L), and DEA/NO (2 to 20 micromol/L). All drugs increased LAD flow and myocardial contractile function in the LAD-dependent myocardium within the first 120 seconds. The greatest inotropic effect was noted after infusion of DEA/NO (20 micromol/L), which increased sWT by 9.7+/-3.1% from 28.5+/-2.2%, Vs by 10.3+/-3.4% from 9.1+/-1.1 mm/s, and RSW by 7.1+/-2.1% from 200.0+/-22.1 mm Hg x mm (P<.05). At the same time, systemic hemodynamics remained unchanged. Prevention of the flow response to GTN by external narrowing of the LAD did not influence the inotropic effect of GTN. CONCLUSIONS: Organic nitrates and NO donors evoke a small but constant positive inotropic effect in vivo that is not caused by coronary vasodilation. PMID- 9355911 TI - Internal cardioversion of atrial fibrillation: marked reduction in defibrillation threshold with dual current pathways. AB - BACKGROUND: The ultimate acceptance of a fully automatic atrial defibrillator will depend on the reduction of pain to acceptable levels, requiring a marked decrease in defibrillation thresholds. The purpose of this study was to determine whether atrial defibrillation thresholds can be reduced by sequential shocks delivered through two current pathways. METHODS AND RESULTS: Sustained atrial fibrillation was induced with rapid atrial pacing in 12 adult sheep. Defibrillation electrodes were positioned in the right atrial appendage (RAap), distal coronary sinus (DCS), proximal coronary sinus (CSos), main/left pulmonary artery junction (PA), and right ventricular apex (RV). Single-capacitor biphasic waveforms (3/1 ms) were delivered through combinations of these electrodes. Probability-of-success curves were determined for single shocks with a single current pathway and sequential shocks with either single- or dual current pathways. The ED50 for delivered energy for the dual current pathway RAap to DCS then CSos to PA was 0.36+/-0.13 J, which was significantly lower than the ED50 of the standard single current pathway RAap to DCS (1.31+/-0.3 J) and was significantly lower than all other configurations tested. CONCLUSIONS: Internal atrial defibrillation thresholds can be markedly reduced with two sequential biphasic shocks delivered over two current pathways compared with the standard single shock delivered over a single current pathway or with sequential shocks delivered over a single current pathway. PMID- 9355912 TI - Beta-adrenergic augmentation of flecainide-induced conduction slowing in canine Purkinje fibers. AB - BACKGROUND: This study was undertaken to test the hypothesis that beta-adrenergic stimulation in the setting of membrane depolarization will potentiate flecainide induced conduction slowing. METHODS AND RESULTS: To elucidate the potential mechanism for the flecainide proarrhythmia observed in CAST, the voltage dependence of beta-adrenergic modulation of impulse propagation in eight flecainide-superfused canine Purkinje fibers was examined with a dual microelectrode technique. At physiological membrane potentials (Vm) ([K+]o=5.4 micromol), 1 micromol flecainide decreased Vmax from 698+/-55 to 610+/-72 V/s (P=.003) and squared conduction velocity (theta2) from 2.11+/-1.1 to 1.72+/-0.9 (m/s)2 (P=.001). With K+ depolarization to Vm=-70 mV, flecainide further reduced Vmax from 306+/-101 to 245+/-65 V/s and theta2 from 1.12+/-0.4 to 0.99+/-0.6 (m/s)2, producing a 2.0-mV hyperpolarizing shift of apparent Na+ channel availability curves derived from theta2. The addition of 1 micromol isoproterenol to flecainide-superfused fibers at physiological Vm increased theta2 by 8% to 1.84+/-0.6 (m/s)2 (P<.01) without altering Vmax. At -70 mV, the addition of isoproterenol magnified the flecainide-induced reduction of Vmax an additional 24% to 185+/-52 V/s (P<.01) and theta2 by 17% to 0.82+/-0.5 (m/s)2 (P=.04), producing an additional 1.8-mV (P=.002) and 1.9-mV (P=.002) hyperpolarizing shift in the apparent Na+ channel inactivation curves generated from Vmax and theta2, respectively. At physiological Vm, the action potential duration (APD95) was reduced from 307+/-35 to 269+/-27 ms (P<.001) by flecainide and subsequently to 217+/-4 ms (P<.001) with isoproterenol addition. With 12 mmol/L K+, APD95 decreased from 198+/-23 to 182+/-17 ms (P=.005) with flecainide and to 164+/-10 ms (P=.004) with isoproterenol. CONCLUSIONS: At depolarized Vm, isoproterenol amplified the flecainide-induced reduction of Vmax and theta2, suggesting a further adrenergic-mediated reduction of Na+ current. Consequently, the synergy between catecholamines and flecainide at depolarized Vm and the shortened APD95 could facilitate arrhythmogenesis in the presence of underlying ischemia. PMID- 9355913 TI - Is epinephrine contraindicated during cardiopulmonary resuscitation? AB - BACKGROUND: Why pulmonary gas exchange deteriorates after administration of epinephrine during cardiopulmonary resuscitation (CPR) is unclear. METHODS AND RESULTS: Forty-four anesthetized swine received an infusion of six inert gases. Animals underwent ventricular fibrillation with CPR and intravenous administration of saline (control), epinephrine (15 microg/kg), or methoxamine (150 microg/kg). Cardiac output, aortic blood pressure, pH, and arterial oxygen saturation were recorded. Distributions of VA and Q were determined by the multiple inert gas elimination technique. Ventricular fibrillation and CPR caused significant decreases in cardiac output, aortic blood pressure, and arterial pH. With epinephrine (versus saline), diastolic blood pressure was significantly higher (23+/-7 versus 8+/-4 mm Hg), but the increase in shunt (from 7+/-4% to 29+/-17%) and the reduction in SaO2 (from 99.7% to 76.8%) were significantly larger. Also, the increase in dead space was greater and elimination of CO2 less. There were no differences between animals given methoxamine or saline, except for increased diastolic blood pressure. CONCLUSIONS: During experimental ventricular fibrillation and CPR, epinephrine increased intrapulmonary shunt approximately 300% more than saline or methoxamine and significantly reduced arterial oxygen saturation. We suspect that the beta-adrenergic receptor activity of epinephrine attenuated hypoxic pulmonary vasoconstriction. Methoxamine is as effective a pressor as epinephrine for CPR and devoid of beta-adrenergic activity. We recommend that such an agent be considered, instead of epinephrine, for CPR. PMID- 9355914 TI - Electrophysiological effects of long, linear atrial lesions placed under intracardiac ultrasound guidance. AB - BACKGROUND: A curative atrial fibrillation procedure will most likely rely on creating transmural linear ablative lesions. However, it is currently unknown whether endocardial radiofrequency lesions can create lines of conduction block. METHODS AND RESULTS: In six pigs, intracardiac echocardiography was used to guide the positioning of multiple coil array catheters to bridge endocardial structures in three right atrial locations: (1) from the crista terminalis to the tricuspid annulus; (2) from the fossa ovalis to the crista terminalis; and (3) from the inferior vena cava to the tricuspid annulus. Once the catheter was positioned, linear lesions were made by radiofrequency energy applied sequentially to each of the four coils. After 15 days, the chest was opened and a 112-electrode epicardial plaque was positioned over the atrial free wall lesion to determine activation patterns. Three lesions were placed in each animal, with a mean procedure time of 47+/-11 minutes. Once adequate contact was determined by intracardiac echocardiography, a single series of radiofrequency application was required to achieve tissue heating (65+/-4 degrees C) with a power of 21+/-10 W. Epicardial mapping demonstrated complete conduction block across the lesions in all animals, with split potentials and disparate activation times (64+/-16 ms) across the lesion. At autopsy, all lesions were discrete, continuous, and without evidence of charring. The lesions were within 0.3+/-0.5 mm of their targeted anatomic locations and measured 21+/-4 mm long and 2.8+/-0.6 mm wide. Histology revealed transmural fibrosis throughout the length of each lesion. CONCLUSIONS: Linear lesions that are electrophysiologically transmural and continuous can be placed in the right atrium of normal pigs. With intracardiac echocardiography, adequate tissue contact over several coil electrodes can be ensured, resulting in short procedure times, efficient energy application, and accurate anatomically linked lesion placement. PMID- 9355915 TI - Hypercholesterolemia exacerbates transplant arteriosclerosis via increased neointimal smooth muscle cell accumulation: studies in apolipoprotein E knockout mice. AB - BACKGROUND: Hypercholesterolemia is thought to be a significant risk factor for coronary vasculopathy in cardiac transplant recipients. METHODS AND RESULTS: We examined the development of arteriosclerosis in mouse carotid artery loops allografted from B.10A(2R) (H-2h2) donors to normocholesterolemic C57BL/6J (H-2h) recipients and hypercholesterolemic C57BL/6J recipients in which the apolipoprotein (apo) E gene had been knocked out. Luminal occlusion and cross sectional neointimal area were greater in arteries allografted into hypercholesterolemic recipients at 15 and 30 days after transplantation. We also measured cellular and extracellular matrix components of the neointima by computerized planimetry of the fractional areas subtended by smooth muscle cells (anti-alpha-actin stain), collagen (Masson's trichrome), lipid (oil red O), and leukocytes (anti-CD45). The neointimal area stained for smooth muscle cells was significantly greater in hypercholesterolemic recipients than in normocholesterolemic recipients at 15 and 30 days after allografting. Lipid contributed to neointimal area to a lesser degree, and there was no significant increase in the contribution of collagen or leukocytes. CONCLUSIONS: Smooth muscle cell accumulation appears to be the principal contributor to the increase in neointimal area observed in arteries allografted into hypercholesterolemic mice. PMID- 9355916 TI - Images in cardiovascular medicine. Renal cell carcinoma with tumor thrombus extending through the inferior vena cava into the right cardiac cavities. PMID- 9355917 TI - Images in cardiovascular medicine. Anomalous origins of the left main coronary artery from the noncoronary sinus and of the right coronary artery from the left sinus of Valsalva. PMID- 9355918 TI - Atrial natriuretic peptide, ventricular myocyte hypertrophy, and hemodynamic overload. PMID- 9355919 TI - How accurately does color kinesis reflect segmental wall motion abnormalities? PMID- 9355920 TI - Congenital heart block and mother's immunology. PMID- 9355921 TI - Contribution of nitric oxide to metabolic coronary vasodilation in the human heart. PMID- 9355922 TI - Aprotinin: topical or systemic delivery in left ventricular assist device implantation. PMID- 9355923 TI - Transesophageal echocardiographic assessment of papillary muscle rupture. PMID- 9355924 TI - Transesophageal echocardiographic assessment of papillary muscle rupture. PMID- 9355925 TI - Methylenetetrahydrofolate reductase (MTHFR) mutation, homocyst(e)ine, and coronary artery disease. PMID- 9355926 TI - Assessment of myocardial viability in patients with chronic coronary disease. PMID- 9355927 TI - Carvedilol in heart failure: the MOCHA trial. PMID- 9355928 TI - Carvedilol and dose-related improvements in left ventricular function. PMID- 9355929 TI - Heart transplantation as a treatment for patients with end-stage Chagas' heart disease. PMID- 9355930 TI - Myocardial endothelin-1 plays a good role and an aggravating role in the failing heart in rats with chronic heart failure. PMID- 9355931 TI - Angiotensin AT1 and AT2 receptor antagonism and myocardial ischemic injury. PMID- 9355932 TI - International Gargellen Conference. PMID- 9355933 TI - Clinical significance of the coronary microcirculation. PMID- 9355934 TI - Aspirin as a therapeutic agent in cardiovascular disease: a statement for healthcare professionals from the American Heart Association. PMID- 9355935 TI - Cataract development induced by repeated oral dosing with FK506 (tacrolimus) in adult rats. AB - FK506 (tacrolimus), a potent immunosuppressant, is used for inhibiting allograft rejection in the organ transplantation field. In a preclinical toxicity study in rats, FK506 induced various toxicities, including renal and pancreatic injuries. One of these toxic findings was cataract, and we have found that cataract appeared in rats dosed orally with FK506 for 13 weeks and more. Therefore, to better elucidate the onset mechanism of FK506-induced cataract, we measured biochemical parameters, such as sorbitol, Na,K-ATPase and glutathione in the lens of rats. Rats were dosed with FK506 in oral daily doses of 0.2, 1 or 5 mg/kg for 13 weeks, the lowest dose of which approximated the expected clinical dosage. Cataract developed in the 5-mg/kg/day group, with an incidence of 25%, whereas no cataract formation was observed in the 0.2- or 1-mg/kg/day groups. Five mg/kg/day led an increase of sorbitol and a decrease of reduced type glutathione, but did not affect Na,K-ATPase activity of the lens. FK506 is known to have diabetogenicity mediated through pancreatic injury, which appears as vacuolation of islet cell in rats. Five mg/kg/day of FK506 induced an elevation of blood glucose associated with glucose intolerance, and decrease of both basal insulin level and insulin content in the pancreas, and the changes were in parallel with the cataract development in the present study. On the other hand, diabetic parameters did not change in the 0.2- or 1-mg/kg/day groups. These observation suggest that diabetes developed in the rats dosed with 5 mg/kg/day of FK506. Coadministration of a novel aldose reductase inhibitor, Zenarestat, at an oral dose of 50 mg/kg/day resulted in a reduction of incidence of the FK506-induced cataract and a decrease of sorbitol levels in the lens when compared to that in the lens of rats dosed with 5 mg/kg/day of FK506. These results suggest that FK506-induced cataract in rats is due to an accumulation of sorbitol in the lens, secondary to the diabetogenic effect of FK506. FK506 treatment at the doses of 0.2 and 1 mg/kg/day neither affected parameters indicative of diabetes nor induced cataract in rats, suggesting that the cataract would not develop with FK506 if diabetic parameters were kept under control. PMID- 9355937 TI - Heat shock protein induction by certain chemical stressors is correlated with their cytotoxicity, lipophilicity and protein-denaturing capacity. AB - Seven agents were analyzed with respect to their ability to induce heat shock protein (HSP) synthesis in C6 rat glioma cells. Induction of HSP synthesis was correlated with cytotoxicity and lipophilicity of the substances. In addition to the first four n-alcohols (methanol, ethanol, propanol and butanol) and phenol, whose capacity to induce HSP was analyzed earlier (Neuhaus-Steinmetz et al., 1994. Mol. Pharmacol. 45, 36-41), isopropanol, 1,4-dinitrophenol (DNP), diethylstilbestrol (DES), carbonylcyanide-m-chlorophenylhydrazone (CCCP), rotenone, paracetamol and acetyl salicylic acid (ASA) induced HSP synthesis after a 1-h incubation at a substance-specific concentration. The maximal induction of HSPs was closely correlated with the cytotoxicity of all substances and occurred when cell viability was reduced to 75 +/- 11% of the controls. Cytotoxicity and the ability to induce HSP were correlated with the lipophilicity of the alcohols, phenol, rotenone and paracetamol. Calculation of the hypothetical membrane concentrations of these compounds yielded a nearly equal value (0.54 +/- 0.13 M), indicating that interaction of substances with lipophilic cellular compounds, such as membranes or lipophilic core regions of proteins, is a critical step leading to HSP induction. This assumption is supported by a correlation between HSP induction and protein denaturation by the different alcohols (Herskovits et al., 1970. J. Biol. Chem. 245, 2588-2598). We assume that the amount of misfolded proteins induced by these lipophilic agents is responsible for the induction of HSP synthesis. ASA, DNP and CCCP induced HSP at lower concentrations than substances with a similar lipophilicity, which may be due to effects which add to the misfolding of proteins or to other signal pathways. PMID- 9355936 TI - Hepatitis A impairs the function of human hepatic CYP2A6 in vivo. AB - Hepatitis virus A (HVA) is a worldwide sporadic disease but its effects on pharmacokinetics and individual drug responses have not been studied. In this study, the 7-hydroxycoumarin (7OHC) excretion test used in vivo as a bioindex of hepatic CYP2A6 activity was performed in 20, previously healthy, acute jaundice HVA patients. Volunteers with an acute HVA were treated with one p.o. administration of 5 mg coumarin (Venalot). Among the patients, 11 were children (6-10 years; two girls and nine boys), the rest (15-40 years old) consisted of two men and seven women. Urinary excretion of 7OHC was measured after overnight fasting in four fractions: 0 h before any medication (to detect if any basal 7OHC excretion exits), and after a 5-mg coumarin capsule p.o., 0-2, 2-4 and 4-8 h fractions were collected and urine volumes were recorded. Urinary excretion of 7 hydroxycoumarin occurred to a similar extent in healthy adults and children. The first 2-h 7OHC excretion was decreased by 26% (P < 0.05) and total (0-8 h) 7OHC excretion was decreased by 37% (P<0.01) among HVA-positive adults (age range 15 40 years) compared with the values obtained from healthy volunteers. In 11 HVA positive children (age 6-10 years), the first 2-h 7OHC excretion was only 20% (P < 0.0001) and the total 7OHC excretion 28% (P < 0.0001) of the value observed in healthy controls. These results suggest that (i) an acute HVA decreases the metabolic clearance of drugs such as coumarin which are rapidly metabolised by CYP2A6 and (ii) this decrease is even more prominent in children. Such metabolic responses may be of clinical importance and may also interfere with other drug therapy in these patients. PMID- 9355938 TI - Reevaluation of cyclosporine induced hepatotoxicity in the isolated perfused rat liver. AB - Livers of male rats were perfused for 120 min in a recirculating hemoglobin-free system with different concentrations of cyclosporine (CS 2, 10, 50, 150 and 200 mg/l). CS produced damage to the livers in a dose dependent manner. The first sign of hepatotoxicity was a reduction of bile flow amounting to 50% already at 50 mg/l CS. At concentrations of 150 mg/l and 200 mg/l, CS lead to a nearly complete suppression of bile flow, furthermore to a release of cytosolic (GPT, glutamate-pyruvate transaminase, LDH, lactate dehydrogenase) and mitochondrial (GLDH, glutamate dehydrogenase) enzymes into the perfusate and to a decrease in hepatic oxygen consumption (30% at 200 mg/l CS). As a consequence of the reduced aerobic energy supply, hepatic ATP concentration declined (70% at 200 mg/l CS). The hepatic concentrations of reduced glutathione (GSH) were not changed but those of oxidized glutathione (GSSG) increased up to 5-fold by CS. Malondialdehyde (MDA) concentrations in the liver and in the perfusate were not affected consistently by CS. The toxic actions of CS in the isolated rat liver were not influenced (a) by the feeding status of the rats (fed or fasted before surgery) or (b) by addition of superoxide dismutase (SOD, 20 mg/l) and catalase (20 mg/l) to the perfusate 30 min before CS. On the other hand, CS-induced hepatic injury could be attenuated or inhibited completely by addition to the perfusate of (1) 2 mmol/l GSH; (2) 12 mmol/l serine; (3) 12 mmol/l glycine; (4) 0.09 mmol/l deferoxamine (DFO). CONCLUSIONS: CS induces cholestasis at lower concentrations, probably by another mechanism(s) than the other signs of hepatotoxicity (enzyme release, ATP depletion). Several lines of evidence indicate a probable participation of reactive oxygen species in CS-induced hepatotoxicity. GSH, DFO, glycine and serine could provide therapeutic opportunities to prevent CS-induced hepatotoxicity in patients treated with high doses of CS. PMID- 9355939 TI - Cyanide induced DNA fragmentation in mammalian cell cultures. AB - Cyanide is a mitochondrial poison and its toxicity is mediated through histotoxic hypoxia. Although cyanide is regarded as a neurotoxin, its other toxic manifestations are also well documented. Cyanide triggers all those events which can lead to DNA damage, but its genotoxic potential has not been established yet. The present investigation addresses the DNA damage induced by cyanide in rat thymocytes in vitro. Cell viability (eosin Y exclusion and LDH leakage) along with DNA strand breaks were measured in thymocytes exposed to 1.25-10 mM KCN for various time intervals. Cleavage into oligonucleosomal fragments of extracted DNA from cyanide treated thymocytes were visualized on gel electrophoresis. Cyanide produced both time and dose dependent DNA fragmentation accompanied by cytotoxicity. The DNA damage was sensitive to elevated levels of extracellular Ca2+ and was minimal in Ca2+ free medium. The DNA fragmentation was attenuated by Zn2+ (modulator of Ca2+/Mg2+-dependent endonuclease), N-acetylcysteine (free radical scavenger) and diltiazem (Ca2+ channel blocker). Cyanide induced DNA damage was further observed in baby hamster kidney cells (BHK-21), where unlike thymocytes, internucleosomal DNA fragmentation was not observed. Thymocytes were more sensitive to cyanide as compared to BHK-21 cells. PMID- 9355941 TI - The role of metabolism in 2-methoxyethanol-induced suppression of in vitro polyclonal antibody responses by rat and mouse lymphocytes. AB - Previous studies from this laboratory have shown that the glycol ether 2 methoxyethanol (ME) produces immunosuppression in the rat but not in the mouse. To investigate possible mechanisms for this species difference in ME-induced immunotoxicity, the effects of ME and its metabolites, 2-methoxyacetic acid (MAA) and 2-methoxyacetaldehyde (MAAD), on in vitro polyclonal antibody responses by Fisher 344 rat and B6C3F1 mouse lymphocytes, were studied. MAAD and MAA suppressed IgM and IgG production by both mouse and rat lymphocytes at non cytotoxic doses. However, ME had no effect on antibody production by either mouse or rat lymphocytes. Lower concentrations of MAA suppressed rat lymphocyte IgM and IgG production (at 0.5 and 1.0 mM MAA, respectively) compared with mouse lymphocytes (2.0 mM MAA). IgM and IgG production by both rat and mouse lymphocytes was suppressed at comparable concentrations of MAAD (0.3 mM MAAD). The role that metabolism of ME to its immunosuppressive forms plays in this in vitro suppression was demonstrated using hepatocyte-lymphocyte co-cultures. IgM production by both mouse and rat lymphocytes was suppressed at a lower concentration of ME following co-culture with mouse (12.5 mM ME) versus rat (25 and 50 mM ME) hepatocytes. These in vitro results indicate that rat lymphocytes are more sensitive to MAA than are mouse lymphocytes and that mouse hepatocytes have a greater capacity to metabolize ME to its immunosuppressive metabolites than do rat hepatocytes. In addition, MAAD is more immunotoxic than MAA, suggesting that this metabolite may be the proximate immunotoxicant. These observation may partially explain the species differences in ME-induced immunosuppression in vivo. PMID- 9355940 TI - Comparison of the effects of di-(2-ethylhexyl)adipate on hepatic peroxisome proliferation and cell replication in the rat and mouse. AB - The effects of di-(2-ethylhexyl)adipate (DEHA) have been compared in female F344 rats and female B6C3F1 mice fed diets containing 0-4.0% DEHA and 0-2.5% DEHA, respectively, for periods of 1, 4 and 13 weeks. In both the rat and mouse treatment with DEHA at all time points produced a dose-dependent increase in relative liver weight and hepatic peroxisome proliferation as demonstrated by the induction of peroxisomal (cyanide-insensitive palmitoyl-CoA oxidation) and microsomal (lauric acid 12-hydroxylase) fatty acid oxidising enzyme activities. The magnitude of induction of peroxisome proliferation was similar in both species. Replicative DNA synthesis was studied by implanting osmotic pumps containing 5-bromo-2'-deoxyuridine during study weeks 0-1, 3-4 and 12-13. After 1 week DEHA treatment hepatocyte labelling index values were increased in rats given 2.5 and 4.0% DEHA and mice given 0.6-2.5% DEHA. While DEHA treatment for 4 and 13 weeks did not increase labelling index values in the rat, a sustained stimulation of replicative DNA synthesis was observed in mice given 1.2 and 2.5% DEHA. The results of this study demonstrate a species difference in the hepatic effects of DEHA, in that at some dose levels DEHA can produce a sustained stimulation of replicative DNA synthesis in mouse but not in rat liver. Sustained cell replication provides a better correlation with the observed formation of liver tumours in chronic studies with DEHA in female mice, but not in female rats, than the magnitude of stimulation of hepatic peroxisome proliferation. PMID- 9355942 TI - Effectiveness of SWL for lower-pole caliceal nephrolithiasis: evaluation of 452 cases. AB - The authors report on the treatment of lower-pole caliceal nephrolithiasis with extracorporeal shockwave lithotripsy (SWL) using the Czech-made Medilit M-5 lithotripter. In 310 patients, they have performed 452 treatments and evaluated the results 3 months after the last session using plain radiographs and ultrasound examination. The effectiveness of treatment and the success rate of SWL decreased with increasing size of stones: with stones >20 mm, it declined to 30%. This poor success rate was attributable not only to the size of the concrement, but also to its location in a lower calix, which is unfavorable for the passage of fragments. For big lower-pole caliceal stones (>20 mm in the longest diameter), the authors recommend percutaneous nephrolithotomy as the primary management method, the effectiveness of which does not depend on the size of the stone. The success rate achieved in treating the lower-pole caliceal lithiasis using the Medilit M-5 machine was 61.3%, similar to that achieved with other lithotripters. PMID- 9355943 TI - Anesthetic management of patients receiving calculus therapy with a third generation extracorporeal lithotripsy machine. AB - We reviewed the anesthetic requirements for satisfactory use of a third generation electromagnetic-source design for extracorporeal shockwave lithotripsy (SWL). Medical records were reviewed for a period of 9 months on all patients receiving anesthesia care for SWL with and without other urologic procedures. The Modulith SL20 was used on 56 ASA Class I-III patients having 87 SWL treatments. Demographic and anesthetic variables were recorded. Complications documented included dysrhythmias, nausea necessitating treatment, and conversion from sedation to regional or general anesthesia. The majority of procedures (83%) were performed on an outpatient basis. Patients were classified as ASA physical status I (27%), II (63%), or III (10%). Monitored anesthesia care with intravenous sedation was utilized in 93% of cases. Of these cases, 78 involved a combination of intravenous propofol, fentanyl, and midazolam; the remaining 3 involved propofol, alfentanil, and/or midazolam. The mean treatment duration was 36 minutes. Patients were discharged within 1 hour after procedure completion in 77 cases (89%). Nausea necessitating treatment was rare (3%). The mean dose of propofol administered with SWL as the only procedure was 272 +/- 112 mg. When SWL was combined with other urologic procedures, the mean dose of propofol was 334 +/ 121 mg. Continuous intravenous propofol infusion provides excellent procedural conditions for SWL on the Modulith SL120, a third-generation lithotripter. PMID- 9355944 TI - Perforation of the bowel during SWL in prone position. AB - Perforation of the small bowel during SWL is reported for the first time. The patient was treated in the prone position for a ureteral stone with 4500 shockwaves. There were no underlying or predisposing factors for the damage to the gut with the exception of the large number of shockwaves at a high energy setting. It is concluded that SWL in the prone position with shockwaves traversing the peritoneal cavity carries a risk of damage to the bowel. A reduction of the number of shockwaves and the energy level should be considered in this setting. PMID- 9355945 TI - Fertility measures in women after extracorporeal shockwave lithotripsy of distal ureteral stones. AB - Long-term effects of extracorporeal shockwave lithotripsy (SWL) on female fertility remain a concern. Thirty-nine women of childbearing age who were treated for distal ureteral stones were surveyed. The mean age of these women was 33 years, and the average stone size was 6.9 mm. The mean calculated radiation exposure to the ovaries and the uterus was 7.53 and 10.9 mSv, respectively. Ten women (26%) attempted to become pregnant. No fertility problems were noted in these women, and 11 healthy babies were delivered. These preliminary findings provide further information regarding the safety of SWL in the treatment of distal ureteral stones in women of reproductive age. PMID- 9355946 TI - High-energy v low-energy shockwave lithotripsy in treatment of ureteral calculi. AB - The size of the crater formed in a urinary calculus subjected to shockwave lithotripsy (SWL) is directly proportional to the energy delivered to the stone surface. This study compared the effect of high and low energy levels on the outcomes of ureteral SWL. Ureteral calculi (N = 336) were treated with the conventional low-energy Siemens Lithostar and 62 with the higher-energy (1.02 v 0.5 mJ/mm2) modification of the Lithostar, the Siemens Shock Tube C. Stone locations included all regions of the ureter. The average stone treated with the standard Lithostar measured 8.1 mm in diameter and received 5461 shockwaves (treatment time 45 minutes) at 17.2 kV (range 14.5-19.0 kV). The stone-free rate was 72%, with stents being used in 16% of patients and a retreatment rate of 9%. The typical stone treated with Shock Tube C was 10.4 mm in diameter and received 3528 shockwaves (treatment time 30 minutes) at an average energy setting of 4.1 (range 1.5-8.0). The stone-free rate was 75%, with stents being used in 9.8% of cases, and a retreatment rate of only 1.6% (P < 0.003). In this study, Shock Tube C yielded stone-free rates equivalent to those of the conventional machine. However, the number of shockwaves, treatment time, and retreatment rate were significantly lower with the new shock tube. High-energy lithotripsy is more efficient than low-energy treatment of ureteral calculi. PMID- 9355947 TI - Effects of extracorporeal shockwave lithotripsy at different stages of pregnancy in the rabbit. AB - Although SWL is now the most common treatment modality for urinary tract stone disease, it is not regarded as a safe method for pregnant patients because of its potential harmful effects on fetus. Using a rabbit model, we investigated whether SWL might cause fetal injury when administered at various developmental stages. Two groups of pregnant rabbits were given 1000 shockwaves either early or late in the gestational period. Time-matched controls did not receive shockwaves. After spontaneous labor, all newborn rabbits were counted, weighted, and measured, and specimens were taken from organs and examined histopathologically. The numbers, weights, and diameters of the newborns in each group were similar. There was no notable histopathologic finding in the heart and brain specimens of any of the newborns, whereas noticeable congestion and multiple focal intraparanchymal microhemorrhages were found in lungs, livers, and kidneys of the animals that had been exposed to shockwaves early in gestation. In conclusion, this study shows that SWL is not a safe treatment in early pregnancy. PMID- 9355948 TI - Ureteroscopic lithotripsy using mini-endoscope and Swiss lithoclast: experience in 147 cases. AB - To evaluate the efficacy of a semirigid mini-endoscope and the Swiss Lithoclast compared with a conventional rigid endoscope and ultrasound, the results of transurethral ureteroscopic lithotripsy in 147 patients over a period of 30 months were analyzed according to the type of ureteroscope (rigid v semirigid) and energy (ultrasound v Lithoclast) used. In the initial 25 cases (Group I), a conventional rigid ureteroscope and ultrasound were used. The latter 122 patients (Group II) were subjected to ureteroscopic lithotripsy using a miniscope and the Lithoclast. The results were superior in Group II with respect to the overall success rate (p = 1.6 x 10[-2]), first-attempt success rate (p = 2.9 x 10[-4]), and the need for ureteral dilation (P = 1.0 x 10[-6]) compared with Group I. There were no major complications. Overall, minor complications (hematuria and urinary tract infection) were observed in 25% of the cases. Further, the results of ureteroscopic lithotripsy in Group II were comparable to those of SWL in situ for upper ureteral calculi and better than for those located in the iliac and lower ureter as reported previously. Our results demonstrate that the Swiss Lithoclast provides effective fragmentation of even hard and smooth stones without increasing the complication rate. This lithotripter is reliable, safe, and simple to operate. In addition, the cost of maintenance is almost nil. PMID- 9355949 TI - Holmium:YAG laser-induced damage to guidewires: experimental study. AB - The holmium:YAG laser fragments stones of all compositions effectively. However, damage to ureteral guidewires by the laser has been described, including in one of our own patients, in whom such damage resulted in morbidity. The purpose of this study was to characterize the interaction of Ho:YAG energy with guidewires in vitro. Seven ureteral guidewires were tested in a waterbath. The 365-microm Ho:YAG laser fiber was placed at defined distances (0, 1, 2, 4, and 5 mm) from the guidewire. All guidewires were tested at angles of 0 degrees, 45 degrees, and 70 degrees from normal incidence. The minimum energy setting that resulted in structural damage to the guidewires was detected by endoscopic video monitoring. All guidewires were susceptible to Ho:YAG laser damage at modest energy settings. The energy required to produce visual damage varied inversely with the square of the distance of the laser fiber from the guidewire. At a distance of 5 mm, none of the guidewires was damaged, even at energy settings of 2.8 J (the maximum output from the laser). The energy required to induce guidewire damage varied with the inverse of the cosine of the incident angle. The results demonstrate that no guidewire is immune from Ho:YAG laser damage when the fiber and guidewire are in contact. Caution must be exercised when operating the Ho:YAG laser near a guidewire, and guidewire integrity should be assured by the surgeon. Generally, the energy required to induce guidewire damage exceeded lithotripsy levels at distances >1 mm and with higher incident angles, implying a reasonable margin of safety during ureteroscopy. The pattern of energy thresholds required to induce damage with respect to distance and incident angle suggests that the mechanism of Ho:YAG lithotripsy is thermal. PMID- 9355950 TI - In vitro and in vivo experiences with ureterorenoscope by Gelet. AB - Using ureterorenoscopes with unidirectional flow, the urologist faces a conflict between a good view on one hand and high pressure in the ureter and the pelvic collecting system of the kidney on the other hand. The ureterorenoscope designed by Gelet, which has continuous (bidirectional) flow and separate irrigation and working channels, was compared with the instrument by Perez-Castro, which has a common irrigation and working access. The features investigated were optical quality, flow-irrigation characteristics, and handling. The latest generation of ureterorenoscopes provides small total diameters because of minimized working channels and fiberoptic image and light transmission. These semirigid instruments combine the features of flexible endoscopes and traditional ureterorenoscopes. The Gelet instrument offers continuous flow, achieved by separation of the irrigation and working channel, and provides a good view even when working instruments are inserted. The additional outlet channel prevents high pressure in the ureter and renal pelvis and offers another access for instrumentation, if necessary, which means maximum safety for the patient. Because the connection of camera and light source on a flexible system is separated from the instrument, ureterorenoscopy is a more convenient procedure for the urologist. PMID- 9355951 TI - Endoscopic treatment of reflux by subureteric collagen injection: critical review of 5 years' experience. AB - In the past decade, subureteric endoscopic injection of Teflon or collagen has been propagated as a safe and successful treatment for vesicoureteral reflux. In our center, from 1990 through 1995, 118 children and 5 adults with reflux and recurrent urinary tract infections were injected with cross-linked bovine collagen in an open, prospective study. Efficacy and safety were assessed 6 and 12 months after injection, and long-term (> or =3 years) results were available in 78 cases. The overall success rate was 58% (64% of ureteral units) free from reflux at 12 months and 54% (58%) after 3 years. Analysis of anatomic, urodynamic, and technical features showed the grade of reflux to be the best predictor of success or failure. When only primary low- and middle-grade reflux, without concomitant anatomic disorders, is considered, the long-term success rate rose to 69% (74% of ureteral units). In conclusion, subureteric collagen injection cannot ultimately replace the highly effective surgical reimplantation. However, the procedure offers a minimally invasive alternative in selected cases of mild reflux when conservative management is inadequate. PMID- 9355952 TI - Contemporary diagnosis and treatment of fibroepithelial ureteral polyp. AB - Fibroepithelial polyps of the ureter are rare, benign tumors often not easily distinguished from malignant transitional-cell carcinomas by radiologic means. Historically, many patients have undergone unnecessary nephroureterectomy for these lesions. With recent advancement in endourologic instrumentation, a biopsy proven diagnosis of suspect upper-tract lesions can be made prior to definitive therapy. We describe a typical case of fibroepithelial ureteral polyp wherein the diagnosis and surgical treatment was accomplished entirely by endoscopic means. PMID- 9355953 TI - Ureteral occlusion prosthesis. AB - Although temporary or definitive complete ureteral occlusion is rarely needed, there is a considerable number of reports introducing different devices to achieve this goal, most of which can be inserted with minimally invasive procedures. Easy placement is considered of paramount importance, as the candidates are very often in bad general condition as a result of previous surgery, radiotherapy, or other palliative treatments for cancer. A device that can be inserted and removed percutaneously is presented herein. It can be employed in cases of ureteral fistulas resulting from radiotherapy and ureterosigmoidostomy with good results. The technique is simple and not time consuming. PMID- 9355954 TI - Laparoscopic management of persistent Mullerian duct remnants associated with an abdominal testis. AB - In recent years, laparoscopy has evolved from a purely diagnostic procedure in the management of nonpalpable testis to a definitive therapeutic intervention. Additional genital malformations occur in association with cryptorchidism, but reports of laparoscopic management of such entities do not exist. Herein, we describe the laparoscopic removal of persistent Mullerian duct remnants (uterus and round ligament) in combination with an orchiectomy of an abnormally small abdominal testis. This technique expands the versatility of laparoscopic management of cryptorchidism to include the resection of associated congenital anomalies. PMID- 9355955 TI - Transperitoneal laparoscopic adrenalectomy: experience in 100 patients. AB - Between July 1992 and October 1996, 100 transperitoneal laparoscopic adrenalectomies were performed on 99 patients at our hospital and affiliated hospitals. The clinical diagnoses were primary aldosteronism (41 patients), Cushing's syndrome (15), pre-Cushing's syndrome (6), pheochromocytoma (7; 8 adrenal glands), adrenal cancer (2), nonfunctioning adenoma (22), myelolipoma (3), and complicated adrenal cyst (3). Ninety-seven glands were removed laparoscopically. The mean operative time was 240 +/- 76 (SD) minutes and the mean blood loss 68 +/- 80 mL for the series. The mean blood was 77 +/- 113 mL when the three operations that were converted to open surgery are included. The mean times for the return to a normal diet and unassisted ambulation were 1.3 +/- 0.6 and 1.4 +/- 0.8 days, respectively. The mean duration of the use of analgesics was 1.5 +/- 1.3 days, including the day of surgery. In contrast, in the latest 10 open adrenalectomies done at Kyoto University Hospital, the mean operative time was 186 +/- 53 minutes and the mean blood loss 220 +/- 170 mL. The mean times for return to a normal diet and for unassisted ambulation and the mean duration of the use of analgesics were 1.9 +/- 0.3, 2.9 +/- 1.1, and 2.9 +/- 1.7 days, respectively. Thirty-six operations, excluding one converted to open surgery, performed at Kyoto University Hospital were selected to look at the learning curve for transperitoneal laparoscopic adrenalectomy and evaluated for operative time and blood loss. The mean operative time and mean blood loss in the first 10 procedures performed at Kyoto University Hospital were 256 +/- 63 minutes and 89 +/- 57 mL; however, these values were reduced to 177 +/- 39 minutes and 48 +/- 32 mL in the next 10 procedures at the same hospital. Laparoscopic adrenalectomy via the transperitoneal anterior approach can be equivalent to open adrenalectomy in efficiency with a shorter convalescence. PMID- 9355957 TI - Transurethral vaporization of ureterocele: case report. AB - A large ureterocele accompanying a bladder stone was vaporized with a large rollerball electrode, permitting electrohydraulic lithotripsy. Conventional endoscopic incision of this lesion had failed. PMID- 9355956 TI - Pilot study of transurethral needle ablation (TUNA) in treatment of nonbacterial prostatitis. AB - Transurethral needle ablation (TUNA) was performed on seven patients with chronic nonbacterial prostatitis who failed to respond to conventional treatments administered for more than half a year. The TUNA procedure heated the prostate to a temperature ranging from 90 degrees to 100 degrees C while the urethral temperature was maintained below 43 degrees C by a protective sheath and irrigation. After treatment, four patients showed complete resolution of symptoms and three a partial improvement. All patients had a decrease in the leukocyte count in expressed prostatic secretions (EPS) 1 month after treatment. Recurrence of abnormal inflammatory cells in the EPS was noted in two patients at 3 months after treatment. The subjective improvement has been maintained during the subsequent follow-up. From these results, TUNA is considered to be an effective, safe, and easy treatment for most patients with nonbacterial prostatitis. PMID- 9355958 TI - Infections in IFNGR-1-deficient children. AB - Human interferon-gamma receptor 1 (IFNGR-1) deficiency is a newly identified autosomal recessive inherited immune disorder. Children with IFNGR-1 deficiency exhibit a severe, profound and selective susceptibility to weakly virulent mycobacteria, such as bacillus Calmette-Guerin (BCG) vaccine or environmental nontuberculous mycobacteria (NTM). This review compares the infections found in IFNGR-1-deficient children to those in IFN-gamma-deficient or IFNGR-1-deficient mice. PMID- 9355959 TI - Quinolinic acid production by macrophages stimulated with IFN-gamma, TNF-alpha, and IFN-alpha. AB - Quinolinic acid (QUIN) has been associated with several inflammatory neurologic disorders, including AIDS dementia complex (ADC). Recent studies suggest that activation of macrophages with either HIV-1 or interferon-gamma (IFN-gamma) can lead to QUIN production. However, the importance of other cytokines, especially those related to the macrophage and that are especially important in ADC pathogenesis, remains unclear. We, therefore, sought to determine the role of tumor necrosis factor-alpha (TNF-alpha) and IFN-alpha in the production of QUIN. Primary human macrophages were stimulated with two different concentrations of these cytokines alone, in combination with each other, and with IFN-gamma. QUIN concentrations in the supernatants were then measured by mass spectrometry at 24, 48, and 72 hs. Results at 72 h showed significant increases in QUIN production in the cells stimulated with IFN-gamma (10297 +/- 170 nmol/L) and also in those stimulated with IFN-alpha (3600 +/- 113 nmol/L), whereas TNF-alpha-stimulated macrophages produced low levels of QUIN (1108 +/- 23 nmol/L). Macrophages stimulated with the cytokine combinations TNF-alpha and IFN-gamma, IFN-alpha, and IFN-gamma, and TNF-alpha and IFN-alpha also resulted in increases in QUIN production (11471 +/- 77.6 nmol/L, 16656 +/- 184 nmol/L, and 3369 +/- 120.5 nmol/L, respectively). The increases in QUIN production in all of the cytokine treatments approached or exceeded in vivo concentrations of QUIN that have been shown to be neurotoxic. These data further support a role for QUIN in cytokine mediated neuronal death in inflammatory disorders of the brain, especially ADC. PMID- 9355960 TI - Interleukin-4 production in Epstein-Barr virus-transformed B cell lines from peripheral mononuclear cells of patients with atopic dermatitis. AB - Interleukin-4 (IL-4) is known as an immunomodulatory cytokine secreted by relatively few cell types, for example, activated T lymphocytes, basophils, and mast cells, but not by B cells. It plays an important role in promoting the production of the IgE antibody. We established Epstein-Barr virus (EBV)-positive B cell lines from peripheral blood mononuclear cells of patients with atopic dermatitis (AD) and tested the production of several cytokines in the cell lines. We found that IL-4 was produced in a cell line, OB, by an IL-4-specific enzyme linked immunosorbent assay. IL-4 mRNA was detected in OB and two other AD-derived cell lines by IL-4-specific reverse transcription-polymerase chain reaction. IgE was also produced by the OB cells. The production of IL-4 and IgE was enhanced in the cells treated with 12-O-tetradecanoyl phorbol-13-acetate. This is the first evidence that IL-4 is produced by an EBV-transformed B cell line. PMID- 9355961 TI - Differential stimulation requirements for IL-12 production among clones of macrophage hybridomas. AB - We have previously established cloned macrophage hybridomas by somatic cell fusion of the macrophage tumor P388D1 of DBA/2 (H-2d) origin with splenic adherent cells of CKB mice (H-2k). Several cloned lines displayed the serologic and functional characteristics of macrophages. In this study, we evaluated the ability of these hybridomas to produce IL-12 after combined stimulation with IFN gamma and lipopolysaccharide (LPS). The patterns of IL-12 production by these cloned macrophages fell into three groups. The first group produced IL-12 on stimulation with LPS in combination with IFN-gamma pretreatment, the second group produced IL-12 on stimulation with LPS regardless of the pretreatment with IFN gamma, and the third group did not produce IL-12 at all on stimulation with IFN gamma and LPS. None of the macrophage clones tested produced IL-12 constitutively. The results correlated well with IL-12 p40 mRNA expression in those macrophages as detected by RT-PCR. These results suggest the differential stimulation requirements for IL-12 production among macrophages at a clonal level. PMID- 9355963 TI - Differential neutralizing activity of human antibodies in interferon antiviral and natural killer cell bioassays. AB - The ability of interferon-alpha (IFN-alpha) to augment the cytotoxicity of human natural killer (NK) cells was used to probe the neutralization capacity of human antibodies to IFN-alpha. Sera from patients treated with IFN-alpha were tested for antibodies that could bind to IFN-alpha, neutralize IFN-alpha antiviral activity, or neutralize IFN-alpha-mediated augmentation of NK cytolytic activity. The NK-augmenting activity of IFN-alpha was measured in a chromium-release cytotoxicity assay using K562 targets and human peripheral blood mononuclear cells in the presence of a constant amount of antibody from patients treated with IFN-alpha. Neutralization of IFN-alpha-mediated antiviral activity did not correlate with neutralization of NK augmentation. However, all sera that neutralized a biologic activity of IFN-alpha also bound IFN-alpha. Conversely, sera that did not bind to IFN-alpha also did not neutralize either biologic activity. The data suggest that some immunogenic epitopes of IFN-alpha reside in distinct domains that mediate different biologic activities of IFN-alpha. The identification of neutralizing antibodies for the NK immunomodulating function of IFN-alpha but not antiviral function suggests that assessment of antiviral neutralization alone may be an incomplete evaluation of the potential significance of binding antibodies that occur subsequent to the administration of therapeutic IFNs. PMID- 9355962 TI - Modulation of cytokine production by a selenoorganic compound (AE-22) in hyperreactive or hyporeactive bronchoalveolar leukocytes of asthmatics or lung cancer patients. AB - We have found that many synthetic selenoorganic compounds, including ebselen, have immunotropic activity. These studies were designed to assess the effect of the analog of ebselen bis[2-pyridyl (2-carbamoyl) phenyl]diselenide (AE-22) on human leukocytes that may express various activation states. The cells were obtained from bronchoalveolar lavage (BAL) cells of patients with various inflammatory lung diseases. The AE-22-treated BAL cells from patients with bronchial asthma (n = 6) and with small cell lung cancer (SCLC) (n = 6) were compared with these in the peripheral blood leukocytes (PBL) from the same donors. The control group comprised 5 patients who underwent diagnostic examination and were free of any cancer or concomitant diseases. Secretion of TNF alpha, IL-6, and IFN-gamma was considered as a marker of BAL or PBL cell activation. Different response of the cells and various effects of AE-22 were observed in relation to the origin and functional state of leukocytes. It was established that AE-22 can induce TNF-alpha, IL-6, and IFN-gamma in a dose dependent manner in BAL cells and PBL isolated from healthy individuals. However, BAL cells were found to be less reactive than PBL as cytokine producers. In contrast, AE-22 had no effect on BAL cells obtained from patients with lung cancer, which were found to be hyporeactive to phytohemagglutinin and bacterial lipopolysaccharide and did not produce TNF-alpha, IL-6, or IFN spontaneously. The spontaneous release of cytokines by BAL cells from bronchial asthma patients, but not by PBL from the same individuals, was significantly (p < 0.01) higher than that from the cultures of healthy control subjects. The high secretion of cytokines by the locally activated BAL cells was significantly (p < 0.01) reduced after administration of AE-22. The results suggest that AE-22 has immunomodulatory activity. AE-22 can downregulate the hyporeactive BAL cells from asthmatics, but it appears to be inactive in BAL cells of cancer patients who can tolerate the cytokine inducers. PMID- 9355964 TI - Low-dose oral type I interferons reduce early virus replication of murine cytomegalovirus in vivo. AB - Immunity to viral infections involves both innate and antigen-specific immune responses. The antiviral properties of interferons (IFNs) are part of the innate immune response. Low doses of type I IFNs (IFN-alpha and IFN-beta) administered daily (10 IU per mouse) by the oral route significantly reduced the early replication of murine cytomegalovirus (MCMV) in both the spleen and liver of MCMV infected susceptible BALB/c mice. Significant inhibition of virus replication was observed for two different inoculum doses of virus (2 x 10(4) pfu per mouse [0.6 LD50] and 2 x 10(4.12) pfu per mouse [0.8 LD50]). Analysis of IFN retention, using [35S]-labeled IFN-alpha1 compared with the nonreceptor binding mutant IFN alpha1 (R33M) administered orally to mice, revealed binding of wild-type IFN alpha1 to several tissues. In particular, IFN was retained by tissues proximal to lymphoid regions, including the posterior nasal cavity, posterior tongue, small intestine, and rectum. These findings suggest that type I IFNs may inhibit MCMV replication by distal binding of the orally administered IFN to various tissues, which in turn augment the primary immune response to virus infection. PMID- 9355965 TI - DNA polymorphisms and mutations of the tumor necrosis factor-alpha (TNF-alpha) promoter in Langerhans cell histiocytosis (LCH). AB - Langerhans cell histiocytosis (LCH) is a clonal proliferation of dendritic histiocytes expressing elevated levels of tumor necrosis factor-alpha (TNF alpha), interferon-gamma (IFN-gamma) granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 (IL-1), and leukemia inhibitory factor (LIF). The cause of the increased cytokine levels is unknown, but DNA sequence changes in promoters could alter expression. The TNF-alpha and IFN-gamma promoter DNA sequences of 12 LCH patients were studied and compared with normal individuals by dideoxy fingerprinting and DNA sequencing. Functional consequences of polymorphic or mutated sequences were assessed by cloning altered and control promoter sequences into a luciferase reporter gene vector. Electrophoretic mobility shifts (EMSA) after binding of nuclear extracts from a macrophage cell line (U-937) by mutated promoters were compared with controls. Five of 12 LCH patients had alterations in the TNF-alpha promoter DNA sequence. None were found in the IFN gamma gene promoter. Of the 5 with TNF-alpha DNA alterations, 2 were at position 308, which has been described as a G-A polymorphism associated with upregulation of TNF-alpha in some patients with infections or immune-mediated diseases. The polymorphism at -308 but not the other TNF-alpha promoter mutations caused a 3 fold to 7-fold increased production of the luciferase reporter gene. EMSA showed that the -308 mutant promoters bound fewer nuclear proteins than normals. Polymorphisms of the TNF-alpha promoter in LCH patients could increase the production of that cytokine. PMID- 9355966 TI - Novel variants of human IFN-alpha detected in tumor cell lines and biopsy specimens. AB - Interferon-alpha constitutes a complex gene family with 14 genes clustered on the short arm of chromosome 9. More than 50 sequence variants have been described. However, an extensive genetic polymorphism has not been seen in the few population studies reported so far. As many of the sequence variants reported were derived from tumor cell lines, we have investigated whether IFN-alpha genes are unstable in tumor cells. Using fluorescence-assisted mismatch analysis (FAMA), combined with allele-specific primer extension, RFLP analysis, and direct sequencing, we detected in a panel of 14 tumor cell lines two new sequence variants of the IFNA1 and IFNA13 genes. Further two-point mutations were found in tumor samples from leukemias (n = 10) and renal cell carcinomas (n = 17) not seen in normal tissues. In the IFNA17 gene, three new sequence variants were detected, one in a tumor cell line and two in tumor biopsy specimens. Besides these individual point mutations, two new polymorphisms were found in each of the IFNA13 and IFNA17 genes. No new variants were found in the IFNA2 and IFNA10 genes. The results suggest that new sequence variants of the IFN-alpha genes occur relatively frequently in tumors or in tumor cell lines. PMID- 9355967 TI - Reversion by deletion of transforming oncogene following interferon-beta and retinoic acid treatment. AB - We have previously shown that prolonged interferon-beta (IFN-beta) treatment of RS485 cells (NIH3T3 cells transformed with multiple copies of an LTR-cHa-ras oncogene) resulted in the phenotypic reversion of 1%-5% of the culture, depending on the conditions used. This reversion persisted after IFN-beta was discontinued, although the revertants retained the LTR-cHa-ras and continued to express ras mRNA and p21. Clones were prepared of such persistent revertant cell lines (PRs). Expression of lysyl oxidase (LOX), which appears to act as a suppressor of ras transformation, was downregulated in RS485 and upregulated in the PRs. When retinoic acid (RA) was combined with IFN-beta treatment of the RS485 cultures, a different mechanism of reversion predominated. Following 60 days of treatment with 20 IU/ml of IFN-beta and 10 microM RA, all of the multiple (3-5) copies of the transforming LTR-c-Ha-ras originally present in RS485 cells were deleted from the genome in 72% of 54 revertant cell lines isolated. As in the case of revertants observed after treatment with IFN-beta alone, LOX mRNA expression was upregulated in all of the revertants that resulted from the treatment with IFN plus RA. The level of LOX mRNA expression acts, therefore, as an indicator of transformation in this system. PMID- 9355968 TI - Expression of hyaluronan in benign and malignant breast lesions. AB - Hyaluronan (HA) is one of the extracellular-matrix components involved in wound healing, tumour growth and metastasis. Due to the limited data on HA expression in benign and malignant breast lesions, we analyzed its presence in these lesions by using the biotinylated-hyaluronan-binding region and the link-protein complex (bHABC) of cartilage proteoglycan as a specific probe. In all benign breast lesions, the expression of HA was restricted to the stromal connective tissue, the ductal epithelial cells being completely devoid of HA. In malignant breast tumours, the intensity of stromal HA staining was significantly stronger than in benign lesions. In addition, HA was detected on cell membranes or in cytoplasms of adenocarcinoma cells, in some cases of ductal carcinoma in situ and in 31% of malignant tumours. The staining pattern was mostly similar in all breast-cancer types studied, i.e., ductal, lobular, tubular, mucinous and medullary. In ductal breast cancer, intense HA expression in stroma and carcinoma cells correlated statistically significantly to poor differentiation of carcinoma, suggesting that altered HA expression may affect the mechanisms of breast-cancer progression. PMID- 9355969 TI - Reduced expression of syndecan-1 in human hepatocellular carcinoma with high metastatic potential. AB - Syndecans comprise a gene family of transmembrane proteoglycans that regulate cellular behavior through interactions with various effectors, including heparin binding growth factors and insoluble matrix components. Syndecan-1, the most extensively studied, localizes in epithelial cells and has been shown to present in normal hepatocytes. However, little is known about the change of syndecan-1 expression in human hepatocellular carcinoma (HCC). We investigated syndecan-1 protein expression by immunohistochemistry in 57 HCC tissue samples. Syndecan-1 gene expression was also determined. Syndecan-1 protein was expressed in cytoplasm and cell membrane of the hepatocytes and in the bile duct epithelial cells of liver with underlying hepatitis and cirrhosis. Conversely, among 57 HCC tissues, 39 HCC (68.4%) showed negative staining; 50% of well-differentiated HCC showed positive staining, whereas 82.4% of poorly differentiated HCC were negative. Loss of syndecan-1-protein expression was more prevalent in HCC with intra-hepatic metastasis (85.2%) than those without metastasis (48.0%). Similarly, syndecan-1 expression was significantly reduced in HCC with extra hepatic metastasis (91.7%) as compared with the HCC without extra-hepatic metastasis (62.2%). The gene expression of syndecan-1 was significantly lower in HCC tissue than that in non-tumoral liver tissue. In 2 human HCC cell lines with poorly differentiated phenotype, HLE and HLF, syndecan-1 expression was markedly decreased both at the mRNA and the protein levels. These results suggest that the loss of syndecan-1 expression is a characteristic feature of HCC with high metastatic potential. PMID- 9355971 TI - Combined analysis of p53 and vascular endothelial growth factor expression in colorectal carcinoma for determination of tumor vascularity and liver metastasis. AB - Recent studies have demonstrated that the p53 tumor suppressor gene plays an important role in controlling tumor angiogenesis. We examined the expression of p53 and vascular endothelial growth factor (VEGF), a well-characterized angiogenic inducer, together with microvessel density to investigate the role of p53 in the regulation of angiogenesis and its clinical significance in human colorectal carcinoma. Surgically resected specimens of 163 colorectal carcinomas were studied by immunohistochemical staining for p53 protein, VEGF and factor VIII-related antigen. Positive p53 protein accumulation and VEGF expression was found in 41.7% and 49.1% of tumors, respectively. p53 and VEGF staining status was identical in 65.6% of tumors. The incidence of p53- or VEGF-positive tumors was significantly higher in patients with venous invasion and liver metastases than in those without. The microvessel count (MVC) in p53- or VEGF-positive tumors was significantly higher than that in negative tumors, and MVC in both p53 and VEGF-positive tumors was significantly higher than that in the other subgroups. Neither synchronous nor metachronous hepatic metastases were found in patients with p53- and VEGF-negative tumors, while 52.2% of patients with both positive tumors had liver metastases and had a poorer prognosis than those with both-negative tumors. Our findings suggest the presence of a p53-VEGF pathway regulating tumor angiogenesis in human colorectal carcinoma. Combined analysis of p53 and VEGF expression might be useful for predicting the occurrence of liver metastasis in patients with this disease. PMID- 9355970 TI - Study of tumor infiltrating lymphocytes and transforming growth factor-beta as prognostic factors in breast carcinoma. AB - Cytokines and growth factors are powerful modulators of the immune response. Their aberrant expression either by the tumor cells or by the tumor infiltrating lymphocytes confers a selective advantage to the tumor to grow and suppress the cytotoxic activity of the infiltrating lymphocytes. Therefore, analysis of these soluble factors in the tumor microenvironment can provide an insight into the understanding of the tumor behavior and may be used as a prognostic factor. In the present study the nature of the tumor infiltrating lymphocytes (TILs) and cytokine profile was examined in 36 and 19 mammary carcinoma tissues, respectively, by immunohistochemistry and PCR. Phenotypic differences in the number of cytotoxic T lymphocytes (CD8+) and lymphokine activated killer cells (CD16) was observed among TILs when patients with either early disease stage (39% and 46.6%, respectively) or those alive with no residual disease (31% and 52%, respectively) were compared with late stage (9.7% and 22.8%, respectively) or those dead of disease (14.6% and 15.6%, respectively). Furthermore, analysis of the 19 tumor samples for cytokine mRNA expression by RT-PCR revealed the presence of TNF-alpha, IL-10, TGF-beta1, and IL-2. However, semi-quantitative PCR analysis demonstrated TGF-beta1 expression to be significantly higher in patients with a favorable outcome (1.0246 attomoles/micromoles) as compared to patients with a poor prognosis (0.1157 attomoles/micromoles). Our results demonstrate the potential biological significance of certain host factors, particularly TILs and TGFbeta1 expression, on the outcome of breast cancer. PMID- 9355972 TI - Prognostic implications of cyclins (D1, E, A), cyclin-dependent kinases (CDK2, CDK4) and tumor-suppressor genes (pRB, p16INK4A) in childhood acute lymphoblastic leukemia. AB - Immunohistochemistry was used to analyze samples of 40 newly diagnosed childhood acute lymphoblastic leukemias (ALL) for their expression of cyclins (D1, E, A), cyclin-dependent kinases (cdk2, cdk4) and tumor-suppressor genes (pRb, p16INK4A), in order to discover whether or not the expression of these various proteins may be of prognostic relevance for the survival of children with ALL. Patients with ALL who were strongly positive for cyclin D1 had a lower probability of remaining in first continuous remission than ALL patients who were negative or weakly positive for this trait. There was also a significant correlation between expression of cyclin D1 and frequency of recurrence. For cyclin E and cyclin A, in contrast, there was no difference in the duration of relapse-free-intervals or the frequency of recurrence in patients. Children with cdk4-positive ALL had a lower probability of remaining in first continuous remission than children with cdk4-negative ALL. No prognostic relevance was found for cdk2. Patients with ALL who expressed pRb had a higher probability and patients who expressed p16 a lower probability of remaining in first continuous remission, but the results were not statistically significant. This investigation demonstrated that cyclin D1 and cdk4 were the most important prognostic factors for children with ALL, and that the combination of them showed the strongest prognostic relevance. PMID- 9355973 TI - Genetic alterations in lobular breast cancer by comparative genomic hybridization. AB - Infiltrating lobular carcinoma (ILC) and infiltrating ductal carcinoma (IDC) are distinguished by their histopathological appearance. However, little is known about the differences in genetic changes between lobular cancers and ductal cancers. We used comparative genomic hybridization (CGH) and compared aberrations in 19 ILCs and 46 IDCs. The total number of aberrations was lower in ILC than in IDC. While the average number of DNA copy number losses did not reach significance between them, copy number gains were significantly lower in ILCs. Fifteen of 19 ILCs (79%) showed increased copy number of 1q, and 12 cases (63%) revealed loss of 16q. The presence of these aberrations was independent of nodal status, histologic subtypes (pleomorphic or classic ILC), or BrdUrd-labeling index. ILCs had a higher frequency of 16q loss than did ductal cancers, and a lower frequency of 8q and 20q gains. Our data suggest that the altered growth pattern and clinical presentation which characterize infiltrating lobular cancers are correlated with distinct genetic alterations. PMID- 9355974 TI - Altered G1 phase regulation in osteosarcoma. AB - Alterations in the normal cell cycle lead to abnormal cell proliferation and to tumor development. To explore the role of the cyclin D/Cdk4 complex and the retinoblastoma protein (pRb) in the growth and spread of osteoblastic osteosarcoma (OS), 40 tumor samples were selected. In 17 of these cases, lung metastases occurred during follow-up. Expression of pRb, cyclin D1 and its catalytic subunit, Cdk4, was studied by immunohistochemistry and immunoblotting. As controls, non-neoplastic tissues surrounding the tumor were used. The expression level and pattern were compared to clinical outcome. Cdk4 was over expressed in 80% of OS, independently of clinical outcome, and showed an intense and uniform distribution in tumor cells compared to normal cells. However, co immunoprecipitation of Cdk4 with cyclin D1 revealed low levels of cyclin D/Cdk4 complex; 20 of 40 OS examined had a negative or minimal immunostaining for active pRb. The probability of relapse was significantly higher in pRb-negative than in the -positive patients (p < 0.05). The ratio of unphosphorylated/hyperphosphorylated pRb was lower in relapsed patients than in patients with no evident disease, though the difference was not statistically significant. High levels of pRb/cyclin D1 were found in all samples exhibiting functional pRb expression. Our results show that G1 phase deregulation is involved in formation and development of OS. The expression levels of both pRb and cyclin D1 had a clear correlation with clinical outcome, suggesting that these parameters could be used as prognostic markers. PMID- 9355975 TI - Decreased expression of alpha-catenin is associated with poor prognosis of patients with localized renal cell carcinoma. AB - During the progression of many cancers, cell-cell adhesion molecules, e.g., E cadherin (EC), may be down-regulated. In a number of carcinomas, EC has been described as an independent prognostic variable. We have studied the expression of adhesion molecules participating in cadherin-catenin complexes in renal cell carcinoma (RCC) specimens. Expression of EC, catenins and p120cas protein was examined in frozen tissue of 90 RCC specimens by immunohistochemistry, and these molecules were evaluated for their significance as prognostic markers. Staining was scored as normal (homogeneously positive at cell-cell borders) or abnormal (heterogeneous or absent). A significant correlation between poor survival and decreased expression of alpha-, beta- or gamma-catenin was observed, whereas no association between survival and EC or p120cas expression was seen. Cox's proportional hazard regression analysis showed that in patients with pT1-3N0M0 disease, reduced alpha-catenin expression correlated with poor survival, suggesting that alpha-catenin expression might be an independent prognostic indicator for patients of this group. PMID- 9355976 TI - p21/waf1/cip1 and mdm-2 expression in breast carcinoma patients as related to prognosis. AB - p21/waf1/cip1 and mdm-2 are downstream effectors of p53. p21 plays a major role in negatively regulating cell-cycle progression, while mdm-2 inhibits p53 effects, and its role has been implicated in oncogenesis. In this study, we investigated the expression profiles of p21, mdm-2 and p53 in human breast carcinoma tissues. The aim was to determine whether a correlation exists between the expression profiles of these markers and tumor differentiation, ER status and prognosis. We studied tumor specimens obtained from 106 patients and found a highly significant association among low histology grade, p53 over-expression, high mdm-2 expression and lack of p21 expression. Our studies also demonstrate that, in human breast cancer, low levels of p21 and higher mdm-2 levels directly correlate with the onset of lymph-node metastases and shortened patient survival. Furthermore, the expression profiles of p21, mdm-2 and p53 were independently correlated with patient survival. PMID- 9355977 TI - Alterations and prognostic significance of p16 and p53 protein expression in subgroups of cutaneous melanoma. AB - The role of p16 and p53 alterations in cutaneous melanoma has been recently discussed, but it remains to be clarified. In the present immunohistochemical study, the expression of p16 and p53 proteins and their possible prognostic relevance have been examined in 102 melanomas of the aggressive nodular type. Twelve percent showed a strong expression of p53 protein, and these cases were significantly more frequent in the head/neck area compared with other sites (32% vs. 6%). Expression of p16 protein was negative or weak in 9% of the cases, and this tended to be less frequent in head/neck tumors compared with the others (0% vs. 12%). Whereas p53 staining was not prognostically important, loss of p16 staining was significantly associated with markedly reduced recurrence free and patient survival in univariate analysis (product-limit method). In multivariate analysis, lack of p16 staining was significantly associated with recurrent disease (p = 0.013). Our findings indicate an important role of altered p16 protein expression in a subgroup of melanoma patients. PMID- 9355978 TI - Detection of disseminated tumor cells in peripheral blood of colorectal cancer patients. AB - All cancer staging systems seek to identify clinical and pathological features that can predict outcome or guide therapy. In particular, a non-invasive method for the early detection of disseminating disease would be of great interest. We investigated the use of cytokeratin genes expression to detect blood metastases from colorectal tumors. Epithelial tumor cells were isolated from whole blood using the monoclonal antibody (MAb) BerEP4 and magnetic beads, and detected by reverse transcription-polymerase chain reaction using oligonucleotides derived from the cDNA sequences of cytokeratins 8, 19 and 20. The sensitivity of this assay was determined by spiking SW620 colon carcinoma cells in normal blood. Using cytokeratin 19 expression we were able to detect 1 epithelial tumor cell in 1 ml of whole blood. The clinical applicability of this technique was explored by evaluating patients with a colorectal carcinoma. Epithelial cells were detected in the blood of 12 out of 23 patients, 2 (20%) of 10 with Astler-Coller stage A or B, and 10 (77%) of 13 with stage C or D cancer. In conclusion, this test is a non-invasive, sensitive, and specific assay for detecting circulating epithelial cells in blood. It may be useful for the early diagnosis of disseminating disease, to determine whether the presence of micrometastatic cells at the time of surgery is correlated with an early relapse and for monitoring adjuvant therapeutic trials. PMID- 9355979 TI - Combined analysis of vascular endothelial growth factor and platelet-derived endothelial cell growth factor expression in gastric carcinoma. AB - Solid tumours require neovascularization for growth and metastasis. Both vascular endothelial growth factor (VEGF) and platelet-derived endothelial cell growth factor (PD-ECGF) are well-characterized inducers of angiogenesis. In this study we examined the expressions of these antigens and their relationship with microvessel density and also determined their prognostic significance. Ninety five specimens resected from patients with gastric carcinoma were investigated using immunohistochemical methods. Microvessel density, determined by immunostaining for factor VIII-related antigen, was significantly higher in tumours that were both VEGF+ and PD-ECGF+ than in tumours that were both VEGF- and PD-ECGF-. According to prognosis, patients with VEGF+ tumours had a significantly worse prognosis than did those with VEGF- tumours. Although there was no significant correlation between PD-ECGF expression and prognosis, patients with PD-ECGF+ tumours tended to have a shorter survival than did those with PD ECGF- tumours. Moreover, the frequency of hepatic recurrence was significantly higher in patients with tumours that were both VEGF-positive and PD-ECGF+ than in all other patients. PMID- 9355980 TI - Features of gastric cancer in hereditary non-polyposis colorectal cancer syndrome. AB - To identify characteristics of gastric cancer associated with hereditary non polyposis colorectal cancer (HNPCC), we gathered clinical data and tumor samples relating to patients recorded in the Finnish HNPCC registry. Our series included 51 families with a characterized mutation and/or that met the Amsterdam criteria. Of 570 members affected by malignancy, gastric cancer occurred in 62. Adequate clinical data were obtained for 45 patients. Tumor samples from 24 patients were re-examined. The mean age of diagnosis of gastric cancer was 56 years. The average percentage of all cancers within a family was 11 (range 0-40). Nineteen were of the intestinal type. Only 3 were of the diffuse type. Helicobacter pylori infection was demonstrated in 3 of 15 cases. Replication error (RER) phenotype was present clearly in 7 cases and at least fairly clearly in 11. The overall 5 year survival rate was 15%. The 5-year survival rate was 48% in cases in whom radical surgery had been undertaken. Our results support the view that gastric cancer belongs to the tumor spectrum of HNPCC. The intestinal type of histology is characteristic, as is the RER+ phenotype, but H. pylori infection was rare. PMID- 9355982 TI - Medical journals turn gaze inward to examine process of peer review. PMID- 9355981 TI - G1 control gene status is frequently altered in resectable non-small cell lung cancer. AB - Progression through the mammalian cell cycle is controlled by a series of cyclins, cyclin-dependent kinases (cdks) and cdk inhibitors. Cyclin D1, cdk4 and the tumour suppressors p16 and retinoblastoma protein (pRb) are thought to comprise a linked system governing cell passage through the G1 phase of the cell cycle. Extending an earlier study on cyclin D1 expression, a series of resectable non-small cell lung carcinomas (NSCLCs) was examined for defects in other elements of this control system. Forty-six of fifty-one NSCLC specimens exhibited at least one alteration of these cell-cycle regulators. Immunohistochemical analysis revealed that 33% and 47% of the tumours failed to express pRb and p16, respectively. Failure to detect pRb did not correlate with loss of heterozygosity at the RB1 locus. Eleven of 12 tumours showing positive (normal) pRb staining over-expressed nuclear localised cyclin D1, including 8 with amplification of the cyclin D1 gene (CCNDI). However, in a number of lesions (n = 5) where cyclin D1 was over-expressed but localised to the cytoplasm, pRb expression was undetectable. Sequencing of exons 1 and 2 of the p16 gene (CDKN2) revealed 3/51 tumours with somatic mutations (in addition to 1 case with a germ-line alteration). All of these lesions were positive for p16 protein. No clear homozygous deletions of CDKN2 were observed by multiplex PCR. As assessed by immunostaining using a p16 monoclonal antibody, there was an inverse correlation of pRb and p16 down-regulation. Whilst patients with tumours over-expressing cyclin D1 had a significantly lower incidence of local relapse, the group whose tumours failed to express pRb had a significantly greater risk of local relapse and tended to have shortened event-free survival. Our data show that alteration of at least one cell cycle-regulator gene occurs in the majority of resectable NSCLCs. PMID- 9355983 TI - Guidelines issued for treatment of ureteral calculi. PMID- 9355984 TI - Assessment of exception to informed consent. PMID- 9355985 TI - From the Food and Drug Administration. PMID- 9355986 TI - From the Centers for Disease Control and Prevention. Self-reported use of mammography among women aged > or =40 years--United States, 1989 and 1995. PMID- 9355987 TI - From the Centers for Disease Control and Prevention. Progress toward global measles control and elimination, 1990-1996. PMID- 9355988 TI - A piece of my mind. Chaos, and the limits of modern medicine. PMID- 9355989 TI - Thrombolytic therapy for elderly patients with myocardial infarction. PMID- 9355990 TI - Thrombolytic therapy for elderly patients with myocardial infarction. PMID- 9355991 TI - Investigational treatments: process, payment, and priorities. PMID- 9355992 TI - Investigational treatments: process, payment, and priorities. PMID- 9355993 TI - Investigational treatments: process, payment, and priorities. PMID- 9355994 TI - Investigational treatments: process, payment, and priorities. PMID- 9355995 TI - Eradicating Legionella from hospital water. PMID- 9355996 TI - 'Z-Pak' vs ice pack: need for clarity and continuous quality assurance. PMID- 9355997 TI - Summertime cluster of intentional ethylene glycol ingestions. PMID- 9355998 TI - Dual effects of weight and weight gain on breast cancer risk. AB - CONTEXT: Breast cancer is a major cause of mortality among women. It is important to identify modifiable risk factors for this disease. OBJECTIVE: To examine body mass index (BMI) at the age of 18 years and at midlife and adult weight change in relation to breast cancer incidence and mortality. DESIGN: Cohort study. SETTING: A cohort of 95256 US female nurses aged 30 to 55 years who were followed up for 16 years. MAIN OUTCOME MEASURE: Incident and fatal breast cancer. RESULTS: During 1203498 person-years, 2517 incident breast cancers (60% postmenopausal) were documented. Higher current BMI was associated with lower breast cancer incidence before menopause and was minimally associated with incidence after menopause. However, a stronger positive relationship was seen among postmenopausal women who never used hormone replacement (relative risk=1.59 for BMI >31 kg/m2 vs < or = 20 kg/m2; 95% confidence interval, 1.09-2.32; P for trend <.001). Higher BMI at the age of 18 years was associated with lower breast cancer incidence both before and after menopause. Weight gain after the age of 18 years was unrelated to breast cancer incidence before menopause, but was positively associated with incidence after menopause. This increased risk with weight gain was limited to women who never used postmenopausal hormones; among these women, the relative risk was 1.99 (95% confidence interval, 1.43-2.76) for weight gain of more than 20 kg vs unchanged weight (P for trend <.001). Current BMI and weight gain were even more strongly associated with fatal postmenopausal breast cancer. In this population, the percentage of postmenopausal breast cancer accounted for by weight gain alone was approximately 16% and by hormone replacement therapy alone was 5%, but when the interaction between these variables was considered, together they accounted for about one third of postmenopausal breast cancers. CONCLUSIONS: Avoiding adult weight gain may contribute importantly to the prevention of breast cancer after menopause, particularly among women who do not use postmenopausal hormones. PMID- 9355999 TI - The relationship between patient income and physician discussion of health risk behaviors. AB - CONTEXT: The US Preventive Services Task Force recommends that physicians assess patients' health risk behaviors, addressing those needing modification. OBJECTIVE: To examine the relationship between patient income, health risk behaviors, the prevalence of physician discussion of these behaviors, and the receptiveness of patients to their physicians' advice. DESIGN: Employee survey. PARTICIPANTS: A random sample of 6549 Massachusetts state employees in 12 health plans. MAIN OUTCOME MEASURES: Data were obtained using a patient-completed mail survey. Trend tests were used to discern differences in the prevalence of health risk behaviors, physician discussion of these behaviors, and patient receptiveness to discussions by patient income. RESULTS: Although unhealthy behaviors were common among all income groups, physician discussion of health risk behaviors fell far short of the universal risk assessment recommended by the US Preventive Services Task Force. Low-income patients were more likely to be obese and smoke than high-income patients and were less likely to wear seat belts and exercise. In contrast, stress and alcohol consumption increased with income, while the proportion of heavy drinkers did not vary significantly. Physicians were more likely to discuss diet and exercise with high-income patients in need of these discussions than with low-income patients, but were more likely to discuss smoking with low-income patients who smoked than with high-income patients who smoked. Among patients with whom discussions occurred, low-income patients were much more likely to report attempting to change their behavior based on physician advice. CONCLUSIONS: Physician counseling of patients regarding health risk behaviors should be greatly improved if the US Preventive Services Task Force recommendations are to be fulfilled. Improvement is especially needed in regard to alcohol consumption, safe sex, and seat belt use. Physicians also need to be more vigilant in properly identifying and counseling low-income patients at risk in regard to diet and exercise and high-income patients who smoke. PMID- 9356000 TI - The cost-effectiveness of air bags by seating position. AB - CONTEXT: Motor vehicle crashes continue to cause significant mortality and morbidity in the United States. Installation of air bags in new passenger vehicles is a major initiative in the field of injury prevention. OBJECTIVE: To assess the net health consequences and cost-effectiveness of driver's side and front passenger air bags from a societal perspective, taking into account the increased risk to children who occupy the front passenger seat and the diminished effectiveness for older adults. DESIGN: A deterministic state transition model tracked a hypothetical cohort of new vehicles over a 20-year period for 3 strategies: (1) installation of safety belts, (2) installation of driver's side air bags in addition to safety belts, and (3) installation of front passenger air bags in addition to safety belts and driver's side air bags. Changes in health outcomes, valued in terms of quality-adjusted life-years (QALYs) and costs (in 1993 dollars), were projected following the recommendations of the Panel on Cost effectiveness in Health and Medicine. PARTICIPANTS: US population-based and convenience sample data were used. MAIN OUTCOME MEASURE: Incremental cost effectiveness ratios. RESULTS: Safety belts are cost saving, even at 50% use. The addition of driver's side air bags to safety belts results in net health benefits at an incremental cost of $24000 per QALY saved. The further addition of front passenger air bags results in an incremental net benefit at a higher incremental cost of $61000 per QALY saved. Results were sensitive to the unit cost of air bag systems, their effectiveness, baseline fatality rates, the ratio of injuries to fatalities, and the real discount rate. CONCLUSIONS: Both air bag systems save life-years at costs that are comparable to many medical and public health practices. Immediate steps can be taken to enhance the cost-effectiveness of front passenger air bags, such as moving children to the rear seat. PMID- 9356001 TI - Effects of aerosolized surfactant in patients with stable chronic bronchitis: a prospective randomized controlled trial. AB - CONTEXT: Chronic bronchitis, estimated to affect more than 13 million adults in the United States, is characterized in part by retention of airway secretions, but no approved or effective therapy for airway mucus retention in patients with chronic bronchitis has been established. Surfactant reduces sputum adhesiveness, which contributes to difficulty in clearing secretions, but surfactant has not been tested in patients with chronic bronchitis. OBJECTIVE: To examine the effects of exogenous surfactant on sputum clearance and pulmonary function in patients with stable chronic bronchitis. DESIGN: A prospective, multicenter, randomized, double-blind, parallel-group, placebo-controlled comparison of the effects of 2 weeks of treatment with 3 doses of aerosolized surfactant (palmitoylphosphadidylcholine [DPPC]) or saline (placebo). SETTING: Four US teaching hospitals. PARTICIPANTS: A total of 87 adult patients with the diagnosis of stable chronic bronchitis. MAIN OUTCOME MEASURES: Pulmonary function, respiratory symptoms, and sputum properties before treatment (day 0), after 2 weeks of treatment (day 14), and 7 days after stopping treatment (day 21). RESULTS: A total of 66 patients were randomized to surfactant treatment and 21 to saline treatment. Patient demographic characteristics between groups were similar at baseline. In patients who received a DPPC dose of 607.5 mg/d for 2 weeks, prebronchodilator forced expiratory volume in 1 second (FEV1) increased from 1.22 L (SEM, 0.08 L) at day 0 to 1.33 L (SEM, 0.09 L) at day 21 (P=.05), an improvement of 11.4%; postbronchodilator FEV1 improved 10.4% by days 14 and 21 (P=.02); and the ratio of residual volume to total lung capacity, a measure of thoracic gas trapping, decreased 6.2% by day 21 (P=.009). In the surfactant groups, there was a dose-dependent increase in the ability of sputum to be transported by cilia in vitro. CONCLUSION: Aerosolized surfactant improved pulmonary function and resulted in a dose-related improvement in sputum transport by cilia in patients with stable chronic bronchitis. PMID- 9356002 TI - Assessing differences in clinical trials comparing surgical vs nonsurgical therapy: using common (statistical) sense. AB - The statement of hypotheses and choice of statistical tests in clinical trials that compare surgical with nonsurgical treatment are complicated by the likelihood of excess risk in the surgical group during the perioperative period but lower risk after that compared with the more uniform risk in the nonsurgical group. Commonly used statistical survival analyses implicitly assume a constant ratio of risks in the 2 groups during the follow-up period. However, the changing pattern of risk for one treatment but not the other implies that the assessment of the relative efficacy of the treatments varies with the length of the follow up. As such, determining whether survival curves for the 2 groups are different may not translate easily into selecting the best treatment. Alternative statements of the hypothesis based on consideration of the time horizon of patients and on clinical judgment may be more consistent with the goals of the study. Regardless of the choice of a statistical test, the choice of treatment is a decision specific to the individual patient and should be influenced by the patient's life expectancy, attitude toward taking risks, quality of life, and cost considerations. When the survival curves cross, there is a trade-off between the risk of surgery and the increase in life expectancy among the survivors of surgery. Accordingly, assessment of differences in outcomes in clinical trials comparing surgical vs nonsurgical therapy should provide both a conclusion about whether 1 treatment can reasonably be considered best for most patients and should provide information to the individual patient and physician on the expected outcome to aid in the decision-making process. PMID- 9356003 TI - Reductions in deaths in frontal crashes among right front passengers in vehicles equipped with passenger air bags. AB - CONTEXT: Virtually all new cars now are equipped with passenger air bags. Determining whether passenger air bags are saving lives is important, particularly because passenger air bags have caused some deaths among children and adults. OBJECTIVE: To assess the effectiveness of passenger air bags in reducing the risk of death in frontal crashes for right front passengers. DESIGN: Air bags are designed to protect occupants in frontal crashes. Using Fatality Analysis Reporting System data for calendar years 1992 through 1995, the relative frequency of right front passenger deaths in frontal vs nonfrontal fatal crashes was compared for cars with dual air bags and for cars with driver-only air bags. MAIN OUTCOME MEASURES: Odds of right front passengers dying in frontal compared with nonfrontal fatal crashes were computed for 1992 through 1995 model year cars with dual air bags and for cars with driver-only air bags. Percentage reductions in right front passenger deaths in dual air bag vehicles were estimated. RESULTS: Right front passenger fatalities were 18% lower than expected in frontal crashes of cars with dual air bags and 11% lower in all crashes. An estimated 73 fewer than expected right front passengers died in 1992 through 1995 model cars with dual air bags during 1992 through 1995. The risk of frontal crash death for right front passengers in cars with dual air bags was reduced 14% among those reported to be using belts and 23% among belt nonusers. Children younger than 10 years in cars with dual air bags had a 34% increased risk of dying in frontal crashes. CONCLUSIONS: Passenger air bags were associated with substantial reductions in fatalities among right front passengers in frontal crashes. However, more children are being killed than are being saved by air bags. Immediate countermeasures to reduce the dangers of air bags to children and adults are suggested. PMID- 9356004 TI - Does this patient have community-acquired pneumonia? Diagnosing pneumonia by history and physical examination. AB - Community-acquired pneumonia is an important cause of acute respiratory symptoms (eg, cough) in the ambulatory care setting. Distinguishing pneumonia from other causes of respiratory illnesses, such as acute bronchitis and upper respiratory tract infections, has important therapeutic and prognostic implications. The reference standard for diagnosing pneumonia is chest radiography, but it is likely that many physicians rely on the patient's history and their physical examination to diagnose or exclude this disease. A review of published studies of patients suspected of having pneumonia reveals that there are no individual clinical findings, or combinations of findings, that can rule in the diagnosis of pneumonia for a patient suspected of having this illness. However, some studies have shown that the absence of any vital sign abnormalities or any abnormalities on chest auscultation substantially reduces the likelihood of pneumonia to a point where further diagnostic evaluation may be unnecessary. This article reviews the literature on the appropriate use of the history and physical examination in diagnosing community-acquired pneumonia. PMID- 9356006 TI - Weight and risk for breast cancer. PMID- 9356005 TI - Wanted: a clearly articulated social ethic for American health care. PMID- 9356007 TI - The next paradigm in health care. PMID- 9356008 TI - Wall Street's role in driving change in health care services. PMID- 9356009 TI - New health care companies: putting physicians in control. PMID- 9356010 TI - The industrialization of health care. PMID- 9356011 TI - Moving GLUT4: the biogenesis and trafficking of GLUT4 storage vesicles. AB - The GLUT4 system in muscle and fat cells plays an important role in whole-body glucose homeostasis. Insulin stimulates the translocation of GLUT4 from an intracellular storage compartment to the cell surface. The nature of this compartment remains largely unknown. We review recent studies describing the biogenesis and molecular constituents of the GLUT4 storage compartment and conclude that it is segregated from the endosomal and biosynthetic pathways. Further, we present evidence to suggest that the GLUT4 storage compartment moves directly to the plasma membrane in response to insulin and, hence, is analogous to small synaptic vesicles in neurons. We propose that the GLUT4 storage compartment be referred to as GLUT4 storage vesicles or GSVs. PMID- 9356012 TI - TNF-alpha-induced insulin resistance in vivo and its prevention by troglitazone. AB - Tumor necrosis factor (TNF)-alpha may play a role in the insulin resistance of obesity and NIDDM. Troglitazone is a new orally active hypoglycemic agent that has been shown to ameliorate insulin resistance and hyperinsulinemia in both diabetic animal models and NIDDM subjects. To determine whether this drug could prevent the development of TNF-alpha-induced insulin resistance, glucose turnover was assessed in rats infused with cytokine and pretreated with troglitazone. Normal male Sprague-Dawley rats were fed normal powdered food with or without troglitazone as a food admixture (0.2%). After approximately 10 days, rats were infused with TNF-alpha for 4-5 days, producing a plasma concentration of 632 +/- 30 pg/ml. In vivo insulin action was measured by the euglycemic-hyperinsulinemic clamp technique at a submaximal (24 micromol x kg[-1] x min[-1]) and maximal insulin infusion rate (240 micromol x kg[-1] x min[-1]). TNF-alpha infusion resulted in a pronounced reduction in submaximal insulin-stimulated glucose disposal rate (GDR) (97 +/- 10 vs. 141 +/- 4 micromol x kg[-1] x min[-1], P < 0.05), maximal GDR (175 +/- 8 vs. 267 +/- 6 micromol x kg[-1] x min[-1], P < 0.01), and in insulin receptor-tyrosine kinase activity (IR-TKA) (248 +/- 39 vs. 406 +/- 32 fmol ATP/fmol IR, P < 0.05). It also led to a marked increase in basal insulin (90 +/- 24 vs. 48 +/- 6 micromol/l, P < 0.05) and free fatty acid (FFA) concentration (2.56 +/- 0.76 vs. 0.87 +/- 0.13 mmol/l, P < 0.01). Troglitazone treatment completely prevented the TNF-alpha-induced decline in submaximal GDR (133 +/- 16 vs. 141 +/- 4 micromol x kg[-1] x min[-1], NS) and maximal GDR (271 +/- 19 vs. 267 +/- 6 micromol x kg[-1] x min[-1], NS). The hyperlipidemia was partially corrected by troglitazone (1.53 +/- 0.28 vs. 0.87 +/- 0.13 mmol/l, P < 0.05), while IR-TKA and insulin concentration remained unaffected by the drug. Troglitazone restores insulin action possibly by lowering the FFA concentration of the blood and/or by stimulating glucose uptake at an intracellular point distal to insulin receptor autophosphorylation in muscle. If TNF-alpha plays a role in the development of the obesity/NIDDM syndrome, troglitazone may prove useful in its treatment. PMID- 9356013 TI - Vanadate activates membranous nonreceptor protein tyrosine kinase in rat adipocytes. AB - The insulin-like effects of vanadate are independent of the insulin receptor and insulin receptor substrate 1 (IRS-1) phosphorylation. A cytosolic protein tyrosine kinase (CytPTK), sensitive to inhibition by nanomolar concentrations of staurosporine (concentration at which 50% inhibition occurs [IC50], 1-2 nmol/l), has been implicated in some (i.e., glucose oxidation, lipogenesis) but not all (i.e., hexose uptake, inhibition of lipolysis) of the insulin-like effects of vanadate. We report here the existence of another nonreceptor protein tyrosine kinase in rat adipocytes, located exclusively in the plasma membranes (MembPTK), which we suggest is associated with hexose uptake and the antilipolytic activity of vanadate. MembPTK is a nonglycoprotein with an estimated molecular weight of 55-60 kDa. In a cell-free experiment, vanadate activates MembPTK seven- to ninefold (median effective dose, 17 +/- 2 micromol/l). Vanadate-activated MembPTK is inhibited by staurosporine (IC50, 60 +/- 5 nmol/l). In intact adipocytes, staurosporine antagonized vanadate-induced hexose uptake (IC50, 6.0 +/- 0.3 micromol/l) and significantly reversed the antilipolytic effect of vanadate (IC50, 5.0 +/- 0.4 micromol/l). After vanadate treatment, a phosphorylated P55 protein is immunoprecipitated by antibodies to both phosphotyrosine and phosphatidylinositol (PI) 3-kinase. In conclusion, rat adipocytes contain an additional vanadate-activatable nonreceptor membranous protein tyrosine kinase that may participate in the effects of vanadate not carried out by CytPTK. We also suggest that after treatment with vanadate, MembPTK is activated by autophosphorylation and interacts with PI 3-kinase. This may explain how vanadate activates PI 3-kinase without involving receptor activation and IRS-1 phosphorylation. PMID- 9356015 TI - Prediction of IDDM in the general population: strategies based on combinations of autoantibody markers. AB - Strategies for assessing risk of progression to IDDM, based on single and combined autoantibody measurement, were evaluated in 2,855 schoolchildren (median age 11.4 years) and 256 children with newly diagnosed IDDM (median age 10.2 years), recruited to a population-based study in the Oxford region. In 256 children with IDDM, levels of antibodies > or =97.5th centile of the schoolchild population were found in 225 (88%) for islet cell antibodies (ICAs), in 190 (74%) for antibodies to GAD, in 193 (75%) for antibodies to protein tyrosine phosphatase IA-2 (IA-2), and in 177 (69%) for autoantibodies to insulin (IAAs). Estimates of risk of progression to IDDM within 10 years, derived by comparing the distribution of antibody markers in the two populations (schoolchildren and children with IDDM), were 6.7% (ICAs), 6.6% (GAD antibodies), 5.6% (IA-2 antibodies), and 4.8% (IAAs) for schoolchildren with levels above the 97.5th centile, increasing to 20, 23, 24, and 11%, respectively, for antibody levels >99.5th centile. Most children with IDDM had multiple antibody markers, and 89% of those diagnosed over age 10 years had > or =2 antibodies above the 97.5th centile, as compared against 0.7% of schoolchildren, in whom this combination gave a 27% 10-year estimated risk of IDDM. Risk increased but sensitivity fell as combined antibody thresholds were raised, or the number of antibodies above the threshold was increased. Strategies based on detection of > or =2 antibodies with primary testing for GAD and IA-2 antibodies and second line testing for ICAs and/or IAAs were evaluated. Detection of at least two markers selected from GAD antibodies > or =97.5th centile and/or IA-2 antibodies > or =99.5th centile and/or ICAs > or =97.5th centile identified 0.25% of schoolchildren and 83% of children with newly diagnosed IDDM, with an estimated risk of 71% (95% CI 57-91). Although confirmation from prospective studies is still needed, this analysis suggests that antibody combinations can predict diabetes in the general population. PMID- 9356016 TI - Cow's milk-free diet does not prevent diabetes in NOD mice. AB - Early dietary exposure to cow's milk proteins has been proposed as an important environmental factor in the development of IDDM both in humans and in diabetes prone rodents. To examine the significance of cow's milk protein in IDDM, 120 NOD mice were maintained, starting from conception until sacrifice, on one of four diets: standard PMI Picolab Rodent Diet 20, a milk-free modification of the standard Picolab diet, a milk-free diet incorporating 0.036% bovine serum albumin (BSA), and a milk-free diet including 0.036% bovine IgG (BGG). The cumulative IDDM incidence at 7 months for these mice in a specific pathogen-free environment on the respective diets was 78, 93, 93, and 67% for females, and 17, 54, 17, and 0% for males. The ages of diabetes onset and insulitis scores were similar for mice on each diet. The unexpectedly lower incidence of IDDM in mice on the milk free diet that included BGG raises the possibility this cow's milk protein might possibly have some protective effect against the development of IDDM in NOD mice. Our main finding was that the standard, milk-free, and BSA-containing diets resulted in comparable incidences of IDDM in NOD mice, demonstrating that neither cow's milk whey proteins in general nor BSA in particular are significantly important as etiologic dietary agents in IDDM in NOD mice. PMID- 9356014 TI - Expression of nitric oxide synthase in skeletal muscle: a novel role for nitric oxide as a modulator of insulin action. AB - Previous studies have shown that nitric oxide synthase (NOS), the enzyme that catalyzes the formation of nitric oxide (NO), is expressed in skeletal muscle. The aim of the present study was to test the hypothesis that NO can modulate glucose metabolism in slow- and fast-twitch skeletal muscles. Calcium-dependent NOS was detected in skeletal muscle, and the enzyme activity was greater in fast type extensor digitorum longus (EDL) muscles than in slow-type soleus muscles. Both the neuronal-type (nNOS) and endothelial-type (eNOS) enzymes are expressed in resting skeletal muscles. However, nNOS protein was only detected in EDL muscles, whereas eNOS protein contents were comparable in soleus and EDL muscles. NOS expression in muscle cryosections (diaphorase histochemistry) was located in vascular endothelium and in muscle fibers, and the staining was greater in type IIb than in type I and IIa fibers. The macrophage-type inducible NOS (iNOS) was not detected in resting muscle, but endotoxin treatment induced its expression, concomitant with elevated NO production. iNOS induction was associated with impaired insulin-stimulated glucose uptake in isolated rat muscles. In vitro, NOS blockade with specific inhibitors did not affect basal or insulin-stimulated glucose transport in EDL or soleus muscles. In contrast, the NO donors GEA 5024 and sodium nitroprusside induced dose-dependent inhibition (up to 50%) of maximal insulin-stimulated glucose transport in both muscles with minor effects on basal uptake values. GEA 5024 also blunted insulin-stimulated glucose transport and amino acid uptake in cultured L6 muscle cells without affecting insulin binding to its receptor. On the other hand, the permeable cGMP analogue dibutyryl cGMP did not affect muscle glucose transport. These results strongly suggest that NO modulates insulin action in both slow- and fast-type skeletal muscles. This novel autocrine action of NO in muscle appears to be mediated by cGMP-independent pathways. PMID- 9356017 TI - Impaired beta-cell function and deposition of fat droplets in the pancreas as a consequence of hypertriglyceridemia in OLETF rat, a model of spontaneous NIDDM. AB - Hypertriglyceridemia is known to be a feature of obesity-related NIDDM, but the patho-etiological significance of this association is obscure. The effects of triglycerides (TGs) on beta-cell function and morphological changes in pancreas were examined using in vivo and in vitro approaches in male OLETF rats at ages 6, 12, and 30 weeks, with their diabetes-resistant counterpart, LETO rats, as normal controls. The results showed that, in the fasting state, plasma TGs in OLETF rats were increased 2.5-fold at age 6 weeks, 3.3-fold at age 12 weeks, and 6.2-fold at age 30 weeks, compared with age-matched LETO rats. The TG content in islets from 12-week-old OLETF rats was significantly increased when compared with those from their age-matched counterparts, but this was not the case with the 6-week-old OLETF rats. Therefore, the islets from 6-week-old rats were cultured with either free fatty acids (FFAs; 1.0 mmol/l sodium oleate) or TG (5.0 mmol/l Intralipide) for 72 h. Several abnormalities in OLETF rats were evident, in contrast to the results from control LETO rats: 1) glucose-induced insulin secretion was more inhibited by either FFAs or TGs in the presence of 27.7 mmol/l glucose, a result associated, at least in part, with reduced glucokinase activity in the islets; 2) a marked elevation in TG content was found in the islets; and 3) the deposition of fat droplets in the enlarged islets, even in the beta-cells, was found by Oil Red O-insulin double staining at age 30 weeks. In conclusion, hypertriglyceridemia resulted in significant TG stores in the islets, which subsequently inhibited glucose-induced insulin secretion, at least in part, via reduced glucokinase activity in the islets. Fat droplets in islets, therefore, may play an important role in hastening the development of NIDDM in this rat model. PMID- 9356018 TI - Release of incompletely processed proinsulin is the cause of the disproportionate proinsulinemia of NIDDM. AB - The production of insulin from proinsulin involves cleavage of intact proinsulin into proinsulin conversion intermediates by the processing of enzymes PC2 and PC3 before fully processed insulin is produced. Intact proinsulin and these conversion intermediates are measured in many immunoreactive insulin (IRI) assays, and therefore contribute to the absolute IRI measurement. The proportion of basal IRI made up of proinsulin (PI)-like molecules (PI/IRI) is increased in NIDDM. Whether stimulated IRI levels are similarly made up of disproportionately increased PI/IRI or whether the relative proportions of proinsulin and its conversion intermediates are altered has not been evaluated. An index of the efficiency of proinsulin processing within the pancreatic beta-cell can be achieved by measuring PI/IRI immediately following acute stimulation of beta-cell secretion, and then determining the proportion of intact proinsulin and proinsulin conversion intermediates contributing to circulating proinsulin-like molecules. In this study, we determined the PI/IRI levels under basal and arginine-stimulated conditions in 17 healthy and 16 NIDDM subjects; high performance liquid chromatography (HPLC) was also performed in a subset of these subjects to measure the relative contribution of intact proinsulin and its conversion intermediates to total proinsulin-like molecules. In NIDDM subjects, levels of both basal (44.6 +/- 9.6 vs. 9.3 +/- 1.5 pmol/l; P = 0.0007) and stimulated (64.0 +/- 12.7 vs. 19.8 +/- 2.8 pmol/l; P = 0.001) proinsulin-like molecules were higher than in healthy subjects. Although IRI was higher in NIDDM than in control subjects under basal conditions (106 +/- 19 vs. 65.1 +/- 8.1 pmol/l; P = 0.05), it was lower in NIDDM than in control subjects following stimulation (increment: 257 +/- 46 vs. 416 +/- 51 pmol/l; P = 0.03). PI/IRI ratios were increased in NIDDM subjects under both basal (43.3 +/- 5.0 vs. 14.0 +/- 1.3%; P < 0.0001) and stimulated (increment: 10.1 +/- 2.1 vs. 2.5 +/- 0.2%; P = 0.0006) conditions, compatible with the release of a disproportionately increased amount of proinsulin-like products. HPLC analysis revealed that, in the stimulated state, intact proinsulin made up 40.1 +/- 6.7% of proinsulin-like molecules in NIDDM individuals (n = 9) and 30.1 +/- 5.6% in healthy subjects (n = 7; NS). The remainder of the proinsulin-like molecules comprised the des-31,32 split proinsulin conversion intermediate. The increase in PI/IRI in NIDDM under basal and especially under stimulated conditions suggests that proinsulin conversion is indeed perturbed in this disorder. Because the relative proportions of intact and des-31,32-split proinsulin are similar in both healthy and NIDDM subjects, the orderly cleavage of proinsulin at its two junctions appears preserved. However, at the time of exocytosis, the secretory granule in the islet of NIDDM subjects contains an increased proportion of incompletely processed proinsulin, presumably reflecting a slower rate of conversion or granules' reduced time of residence in beta-cells. PMID- 9356019 TI - Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin-producing cells. AB - Antioxidant enzyme expression was determined in rat pancreatic islets and RINm5F insulin-producing cells on the level of mRNA, protein, and enzyme activity in comparison with 11 other rat tissues. Although superoxide dismutase expression was in the range of 30% of the liver values, the expression of the hydrogen peroxide-inactivating enzymes catalase and glutathione peroxidase was extremely low, in the range of 5% of the liver. Pancreatic islets but not RINm5F cells expressed an additional phospholipid hydroperoxide glutathione peroxidase that exerted protective effects against lipid peroxidation of the plasma membrane. Regression analysis for mRNA and protein expression and enzyme activities from 12 rat tissues revealed that the mRNA levels determine the enzyme activities of the tissues. The induction of cellular stress by high glucose, high oxygen, and heat shock treatment did not affect antioxidant enzyme expression in rat pancreatic islets or in RINm5F cells. Thus insulin-producing cells cannot adapt the low antioxidant enzyme activity levels to typical situations of cellular stress by an upregulation of gene expression. Through stable transfection, however, we were able to increase catalase and glutathione peroxidase gene expression in RINm5F cells, resulting in enzyme activities more than 100-fold higher than in nontransfected controls. Catalase-transfected RINm5F cells showed a 10-fold greater resistance toward hydrogen peroxide toxicity, whereas glutathione peroxidase overexpression was much less effective. Thus inactivation of hydrogen peroxide through catalase seems to be a step of critical importance for the removal of reactive oxygen species in insulin-producing cells. Overexpression of catalase may therefore be an effective means of preventing the toxic action of reactive oxygen species. PMID- 9356020 TI - A nonsense mutation in the inward rectifier potassium channel gene, Kir6.2, is associated with familial hyperinsulinism. AB - ATP-sensitive potassium (K[ATP]) channels are an essential component of glucose dependent insulin secretion in pancreatic islet beta-cells. These channels comprise the sulfonylurea receptor (SUR1) and Kir6.2, a member of the inward rectifier K+ channel family. Mutations in the SUR1 subunit are associated with familial hyperinsulinism (HI) (MIM:256450), an inherited disorder characterized by hyperinsulinism in the neonate. Since the Kir6.2 gene maps to human chromosome 11p15.1 (1,2), which also encompasses a locus for HI, we screened the Kir6.2 gene for the presence of mutations in 78 HI probands by single-strand conformation polymorphism (SSCP) and nucleotide sequence analyses. A nonsense mutation, Tyr- >Stop at codon 12 (designated Y12X) was observed in the homozygous state in a single proband. 86Rb+ efflux measurements and single-channel recordings of COS-1 cells co-expressing SUR1 and either wild-type or Y12X mutant Kir6.2 proteins confirmed that K(ATP) channel activity was abolished by this nonsense mutation. The identification of an HI patient homozygous for the Kir6.2/Y12X allele affords an opportunity to observe clinical features associated with mutations resulting in an absence of Kir6.2. These data provide evidence that mutations in the Kir6.2 subunit of the islet beta-cell K(ATP) channel are associated with the HI phenotype and also suggest that the majority of HI cases are not attributable to mutations in the coding region of the Kir6.2 gene. PMID- 9356021 TI - Diminished insulin and glucagon secretory responses to arginine in nondiabetic subjects with a mutation in the hepatocyte nuclear factor-4alpha/MODY1 gene. AB - Nondiabetic subjects with the Q268X mutation in the hepatocyte nuclear factor (HNF)-4alpha/MODY1 gene have impaired glucose-induced insulin secretion. To ascertain the effects of the nonglucose secretagogue arginine on insulin and glucagon secretion in these subjects, we studied 18 members of the RW pedigree: 7 nondiabetic mutation negative (ND[-]), 7 nondiabetic mutation positive (ND[+]), and 4 diabetic mutation positive (D[+]). We gave arginine as a 5-g bolus, followed by a 25-min infusion at basal glucose concentrations, and after glucose infusion to clamp plasma glucose at approximately 200 mg/dl. The acute insulin response (AIR), the 10-60 min insulin area under the curve (AUC), and the insulin secretion rate (ISR) were compared, as were the acute glucagon response (AGR) and glucagon AUC. The ND[+] and D[+] groups had decreased insulin AUC and ISR and decreased glucose potentiation of AIR, insulin AUC, and ISR to arginine administration when compared with the ND[-] group. At basal glucose concentrations, glucagon AUC was greatest for the ND[-] group, intermediate for the ND[+] group, and lowest for the D[+] group. During the hyperglycemic clamp, there was decreased suppression of glucagon AUC for both ND[+] and D[+] groups compared with the ND[-] group. The decreased ISR to arginine in the ND[+] group compared with the ND[-] group, magnified by glucose potentiation, indicated that HNF-4alpha affects the signaling pathway for arginine-induced insulin secretion. The decrease in glucagon AUC and decreased suppression of glucagon AUC with hyperglycemia suggest that mutations in HNF-4alpha may lead to alpha-cell as well as beta-cell secretory defects or a reduction in pancreatic islet mass. PMID- 9356022 TI - GLP-I(7-36) amide augments Ba2+ current through L-type Ca2+ channel of rat pancreatic beta-cell in a cAMP-dependent manner. AB - The whole-cell patch-clamp method was used to examine the effect of glucagon-like peptide I (GLP-I)(7-36) amide on the activation process of L-type Ca2+ channels of rat pancreatic beta-cells. After depolarization, GLP-I (1-100 nmol/l) caused action potentials in cells exposed to a glucose-free solution for 10 min. The percentage of cells producing action potential depended on the concentration of GLP-I. In some cells, GLP-I caused action potentials without the prior depolarization of the membrane. In cells exposed to the glucose-free solution for longer than 30 min, or in cells that were deprived of ATP by a means of the conventional whole-cell configuration, GLP-I (20 nmol/l) did not cause the electrical excitation. Application of GLP-I augmented the maximum Ba2+ current (IBa) through L-type Ca2+ channels and shifted the current voltage curve to the left. Values of changes in the maximum IBa depended on GLP-I concentration. Application of dibutyryl cAMP (dbcAMP, 1 mmol/l) also augmented IBa. In cells pretreated with Rp-cAMP, dbcAMP did not change the magnitude of IBa. Also in cells pretreated with Rp-cAMP, GLP-I failed to augment IBa. These results suggest that in pancreatic beta-cells, GLP-I, by a cAMP-dependent mechanism, increases opening of L-type Ca2+ channels. cAMP-dependent augmentation of Ca2+ entry as well as cAMP production itself by GLP-I plays a crucial role in controlling insulin secretion. PMID- 9356023 TI - Insulin resistance of muscle glucose transport in rats fed a high-fat diet: a reevaluation. AB - Rats fed a high-fat diet develop skeletal muscle insulin resistance. There is disagreement regarding whether a decrease in the GLUT4 isoform of the glucose transporter is responsible. We found that feeding rats a high-fat diet that reduced the responsiveness of glucose transport to insulin in skeletal muscles by approximately 25-45% in 4 weeks, had no significant effect on muscle GLUT4 content. There is also controversy regarding whether the contraction/anoxia activated pathway of glucose transport stimulation is affected by fat feeding. We found that stimulation of muscle glucose transport by either swimming, in situ contractions, or anoxia was depressed to a similar extent as insulin responsiveness in high-fat-fed rats. It has been suggested that the muscle insulin resistance caused by a high-fat diet is due to increased fat oxidation and glucose-fatty acid cycle activity. However, we found that insulin-stimulated glucose transport was reduced by approximately 40% when muscles of fat-fed rats were incubated under anoxic conditions under which fatty acid oxidation should not occur. Rats maintained on the high-fat diet up to 32 weeks developed the characteristics of the abdominal obesity syndrome, including insulin resistance, hyperinsulinemia, hyperglycemia, elevated LDL cholesterol and VLDL triglycerides, and marked visceral obesity. We conclude that 1) in rats fed a high-fat diet the muscle insulin resistance is not due to a decrease in total GLUT4 content or to increased fat oxidation, 2) fat feeding also results in resistance of muscle glucose transport to stimulation via the contraction/anoxia pathway, and 3) rats fed a high-fat diet may be a useful model of the abdominal obesity syndrome. PMID- 9356024 TI - Mechanisms of liver and muscle insulin resistance induced by chronic high-fat feeding. AB - To elucidate cellular mechanisms of insulin resistance induced by excess dietary fat, we studied conscious chronically high-fat-fed (HFF) and control chow diet fed rats during euglycemic-hyperinsulinemic (560 pmol/l plasma insulin) clamps. Compared with chow diet feeding, fat feeding significantly impaired insulin action (reduced whole body glucose disposal rate, reduced skeletal muscle glucose metabolism, and decreased insulin suppressibility of hepatic glucose production [HGP]). In HFF rats, hyperinsulinemia significantly suppressed circulating free fatty acids but not the intracellular availability of fatty acid in skeletal muscle (long chain fatty acyl-CoA esters remained at 230% above control levels). In HFF animals, acute blockade of beta-oxidation using etomoxir increased insulin stimulated muscle glucose uptake, via a selective increase in the component directed to glycolysis, but did not reverse the defect in net glycogen synthesis or glycogen synthase. In clamp HFF animals, etomoxir did not significantly alter the reduced ability of insulin to suppress HGP, but induced substantial depletion of hepatic glycogen content. This implied that gluconeogenesis was reduced by inhibition of hepatic fatty acid oxidation and that an alternative mechanism was involved in the elevated HGP in HFF rats. Evidence was then obtained suggesting that this involves a reduction in hepatic glucokinase (GK) activity and an inability of insulin to acutely lower glucose-6-phosphatase (G-6-Pase) activity. Overall, a 76% increase in the activity ratio G-6-Pase/GK was observed, which would favor net hepatic glucose release and elevated HGP in HFF rats. Thus in the insulin-resistant HFF rat 1) acute hyperinsulinemia fails to quench elevated muscle and liver lipid availability, 2) elevated lipid oxidation opposes insulin stimulation of muscle glucose oxidation (perhaps via the glucose-fatty acid cycle) and suppression of hepatic gluconeogenesis, and 3) mechanisms of impaired insulin-stimulated glucose storage and HGP suppressibility are not dependent on concomitant lipid oxidation; in the case of HGP we provide evidence for pivotal involvement of G-6-Pase and GK in the regulation of HGP by insulin, independent of the glucose source. PMID- 9356025 TI - Insulin signaling in human skeletal muscle: time course and effect of exercise. AB - Activation of early steps in the insulin signaling cascade in human skeletal muscle was investigated using a one-step euglycemic-hyperinsulinemic (approximately 100 pU/ml) clamp in seven healthy young men 3 h after one-legged exercise. Concomitant insulin stimulation (three- to six-fold [P < 0.05]) of thigh glucose clearance, muscle insulin receptor tyrosine kinase (IRTK), insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation, and IRS-1-associated phosphatidylinositol 3-kinase (PI 3-kinase) was observed in the rested leg. Twenty minutes after cessation of insulin infusion, the level of these parameters returned toward basal. A twofold higher insulin-stimulated glucose clearance in the exercised compared with the rested thigh was accompanied by unaltered maximal IRTK activation and IRS-1 tyrosine phosphorylation, and by a decreased (approximately 50%, P < 0.05) maximal IRS-1 associated PI 3-kinase activation. Prior exercise caused significantly faster insulin-stimulated tyrosine phosphorylation of IRS-1, PI 3-kinase activity, and glucose clearance compared with those in the rested thigh. In conclusion, physiological hyperinsulinemia activates IRTK, IRS-1 tyrosine phosphorylation, and PI 3-kinase in human skeletal muscle. However, increased insulin action after exercise is not caused by potentiation of these steps in the insulin signaling cascade. In contrast, at steady state, paradoxically decreased insulin-stimulated IRS-1-associated PI 3 kinase activity was observed in exercised muscle. Thus, the activity of IRS-1 associated PI 3-kinase and glucose uptake may not always be tightly coupled during insulin stimulation in human muscle. PMID- 9356026 TI - Inhibition of food response to intracerebroventricular injection of leptin is attenuated in rats with diet-induced obesity. AB - The fat-derived hormone, leptin, is thought to regulate adipose tissue mass by acting on the brain to reduce food intake and increase thermogenesis. We have produced obesity in rats more than 8 weeks old by feeding a high-calorie diet and have then examined the inhibitory effect of intracerebroventricularly injected recombinant murine leptin on their food intake versus control rats. In control rats, randomized injections of leptin (0.5, 2.0, or 10.0 microg) or sterile saline vehicle into the lateral ventricle produced a dose-dependent reduction in normal laboratory diet consumed 1, 4, and 24 h after the lights were turned off. However, in diet-induced obesity, the dose-dependent inhibition of food intake was observed at 1 h only, and the effect was attenuated. Switching the diet induced obese rats to a normal laboratory diet 1 week before injections of leptin were commenced resulted in a reduction in the daily food consumption. These data suggest that rats made obese by feeding a high-calorie diet override the normal satiety effects of leptin since when they are returned to a normal laboratory diet, they reduce their calorie intake, possibly as a result of a restoration of the satiety effects of endogenous leptin. However, the fact that the hypophagic response to exogenous leptin is impaired in these rats at this time suggests some residual impairment of the satiety signal, perhaps caused by reduced receptor sensitivity and/or near total occupation of receptors by endogenous leptin molecules, levels of which are raised in plasma. PMID- 9356028 TI - Inhibition of food intake by neuropeptide Y Y5 receptor antisense oligodeoxynucleotides. AB - The recently discovered rat neuropeptide Y (NPY) receptor, the Y5 subtype, has been proposed to mediate the NPY-induced feeding response and therefore plays a central role in the regulation of food intake. These conclusions were based on studies with peptidic agonists. We now report studies in which phosphothioate end protected antisense oligodeoxynucleotides (ODNs) targeted to prepro NPY (prepro NPY antisense ODNs) or to the Y5 receptor (Y5 antisense ODNs) were used to assess the functional importance of this novel receptor subtype in vivo. NPY antisense ODNs given intracerebroventricularly to rats prevented the increase in hypothalamic NPY levels during food deprivation and inhibited fasting-induced food intake. Likewise, repeated intracerebroventricular injections of Y5 antisense ODNs prevented fasting-induced food intake in rats. Moreover, two Y5 antisense ODNs, targeted to different sequences of the receptor, significantly decreased basal food intake and inhibited the increase in food intake after intracerebroventricular injection of NPY. These effects proved to be selective, since the feeding response to galanin was not affected. Analysis of the structure of feeding behavior revealed that prepro NPY and Y5 receptor antisense ODNs reduced food intake by inducing decreases in meal size and meal duration analogous to the orexigenic effects of NPY that are mediated by increases in these parameters. Although changes in Y5 receptor density could not be measured, the results with Y5 antisense ODNs strongly suggest that this receptor subtype mediates the feeding response to exogenous and endogenous NPY. Selective Y5 antagonists may therefore be of therapeutic value for the treatment of obesity and eating disorders. PMID- 9356027 TI - Elevated intakes of supplemental chromium improve glucose and insulin variables in individuals with type 2 diabetes. AB - Chromium is an essential nutrient involved in normal carbohydrate and lipid metabolism. The chromium requirement is postulated to increase with increased glucose intolerance and diabetes. The objective of this study was to test the hypothesis that the elevated intake of supplemental chromium is involved in the control of type 2 diabetes. Individuals being treated for type 2 diabetes (180 men and women) were divided randomly into three groups and supplemented with: 1) placebo, 2) 1.92 micromol (100 microg) Cr as chromium picolinate two times per day, or 3) 9.6 micromol (500 microg) Cr two times per day. Subjects continued to take their normal medications and were instructed not to change their normal eating and living habits. HbA1c values improved significantly after 2 months in the group receiving 19.2 pmol (1,000 microg) Cr per day and was lower in both chromium groups after 4 months (placebo, 8.5 +/- 0.2%; 3.85 micromol Cr, 7.5 +/- 0.2%; 19.2 micromol Cr, 6.6 +/- 0.1%). Fasting glucose was lower in the 19.2 micromol group after 2 and 4 months (4-month values: placebo, 8.8 +/- 0.3 mmol/l; 19.2 micromol Cr, 7.1 +/- 0.2 mmol/l). Two-hour glucose values were also significantly lower for the subjects consuming 19.2 micromol supplemental Cr after both 2 and 4 months (4-month values: placebo, 12.3 +/- 0.4 mmo/l; 19.2 micromol Cr, 10.5 +/- 0.2 mmol/l). Fasting and 2-h insulin values decreased significantly in both groups receiving supplemental chromium after 2 and 4 months. Plasma total cholesterol also decreased after 4 months in the subjects receiving 19.2 micromol/day Cr. These data demonstrate that supplemental chromium had significant beneficial effects on HbA1c, glucose, insulin, and cholesterol variables in subjects with type 2 diabetes. The beneficial effects of chromium in individuals with diabetes were observed at levels higher than the upper limit of the Estimated Safe and Adequate Daily Dietary Intake. PMID- 9356029 TI - Interstitial muscle insulin and glucose levels in normal and insulin-resistant Zucker rats. AB - To study interstitial insulin and glucose concentrations, microdialysis was performed in the medial femoral muscles in normal SD rats as well as in insulin resistant obese Zucker rats during a euglycemic insulin clamp. [14C]inulin was given (0.1 mCi/rat) as a constant subcutaneous infusion 24 h before the insulin clamp. Insulin infusion rates were 5-8 mU x kg(-1) x min(-1) (low rate) for 140 min and 10-20 mU x kg(-1) x min(-1) (high rate) for another 100 min. The relationship between insulin and [14C]inulin dialysate recoveries was evaluated in vivo and in vitro in plasma to calculate interstitial insulin concentration. Relative microdialysis recovery of interstitial insulin in vivo was 3.0 +/- 0.3% (mean +/- SE, n = 68). In normal SD rats, plasma and interstitial insulin concentrations were identical when plasma insulin was < or =250 mU/ml, whereas interstitial insulin was lower when plasma insulin was > or =350 mU/ml. Half maximal glucose infusion rate was achieved in the presence of plasma and interstitial insulin concentrations of approximately 140 mU/ml, whereas maximal glucose disposal was seen at interstitial insulin concentrations of approximately 325 mU/ml, corresponding to approximately 500 mU/ml in plasma. In electrically stimulated and contracting (1 Hz) normal muscle with markedly increased blood flow, the dialysate insulin concentration was significantly higher at high rates, but not at low rates, of insulin infusion. In insulin-resistant obese Zucker rats, the interstitial insulin concentration was similar to that in plasma, even at pharmacological concentrations. The glucose infusion rate was significantly lower in the obese Zucker rats at both insulin infusion rates than in the lean animals. The glucose content in dialysates from skeletal muscle was equal in both obese and lean rats during the low insulin infusion rate. During the high insulin infusion rate, dialysate glucose concentrations decreased significantly in both groups but were significantly higher in the obese Zucker rats. The data suggest that transport of insulin and glucose diffusion across the capillary wall are rate limiting for insulin as well as for glucose metabolism in muscle in normal rats. This does not appear to be the case in the insulin-resistant obese Zucker rats, where the reduced insulin responsiveness in muscle is due to muscular cellular defects rather than an inhibited transcapillary delivery of insulin. PMID- 9356030 TI - Insulin-independent acute restoration of euglycemia normalizes the impaired glucose clearance during exercise in diabetic dogs. AB - At rest and during exercise, chronic hyperglycemia, high free fatty acid (FFA) oxidation, and insulin deficiency in diabetes are well known to impair glucose clearance (metabolic clearance rate [MCR]). The effect of acute restoration of glycemia per se on MCR has been less well characterized. We therefore studied normal and alloxan-diabetic dogs both at rest and during exercise, as diabetic hyperglycemic or after acutely induced euglycemia (<160 min) generated by infusion of either insulin or phlorizin. Glucose uptake was similar under hyperglycemic and normoglycemic conditions both at rest and during exercise, indicating a precise balance between the mass effect of glucose and decreased MCR. Rest and exercise MCR was fourfold lower under conditions of hyperglycemia, but insulin-independent restoration of euglycemia improved basal MCR threefold and normalized MCR during exercise. High FFA turnover did not affect glucose uptake but was correlated with plasma lactate concentrations (r = 0.72, P < 0.001), suggesting that muscle fuel requirements are controlled by glucose oxidation and not uptake. We conclude that in alloxan-diabetic dogs, the impaired MCR may be an adaptive phenomenon because correction of hyperglycemia corrects MCR despite partial insulin deficiency and high FFA turnover. We speculate that constant glucose uptake despite hyperglycemia in diabetes may protect the muscle from excessive exposure to glucose. PMID- 9356031 TI - Reassessment of glucose effectiveness and insulin sensitivity from minimal model analysis: a theoretical evaluation of the single-compartment glucose distribution assumption. AB - Minimal model analysis with the frequently sampled intravenous glucose tolerance test provides an effective way to measure two important metabolic parameters in vivo under non-steady-state conditions: glucose effectiveness (SG) and insulin sensitivity (SI). Two questions regarding the validity of SG and SI have recently emerged. First, SG from the minimal model is suspected to be overestimated. Second, the occurrence of SI values indistinguishable from zero ("zero-SI") is not negligible in large clinical studies, and its physiological meaning is uncertain. In this study, we examined the significance of the assumed single compartment glucose distribution embedded in the minimal model on the estimation of SG and SI. A more accurate two-compartment model was constructed by incorporating insulin action on hepatic glucose output and uptake into a previously validated construction. The two-compartment results were compared with the one-compartment minimal model results. It was shown that the one-compartment assumption contributes to a systematic deviation of SG (slope = 0.54, y-intercept = 0.014 min[-1]; n = 195 simulations). However, SG from the minimal model was linearly correlated to SG determined from the two-compartment model (r = 0.996). The one-compartment assumption also contributed to the occurrence of zero SI values for insulin-resistant subjects. A similar linear relationship was found between SI estimated by both the minimal model and the two-compartment model (slope = 0.58, y-intercept = -0.57 x 10[-4] min[-1] per pU/ml, r = 0.998). In conclusion, SG and SI from the minimal model are not necessarily equivalent to values emanating from the more accurate two-compartment model. However, the very high correlation between one- and two-compartment results suggests that the minimal model-derived SG and SI are dependable indexes of in vivo glucose effectiveness and insulin sensitivity. Minimal model analysis' advantages of simplicity, minimal invasiveness, reasonable reflection of non-steady-state glucose kinetics, and cost-effectiveness could in many cases outweigh the structural bias introduced by the model simplification. PMID- 9356032 TI - Evidence of an increased number of type IIb muscle fibers in insulin-resistant first-degree relatives of patients with NIDDM. AB - Insulin resistance is a common feature in first-degree relatives of NIDDM patients. To explore the mechanism(s) behind this condition in more detail, a percutaneous muscle biopsy (vastus lateralis) was performed in 25 first-degree relatives of NIDDM patients and 21 control subjects to examine muscle fiber composition and capillary density. Insulin-stimulated glucose disposal (Rd) was determined employing a hyperinsulinemic-(insulin infusion rate 0.6 mU x kg[-1] x min[-1]) euglycemic clamp. Rd (5.76 +/- 0.35 vs. 8.06 +/- 0.36 mg x kg lean body weight [LBW]-1 x min[-], P < 0.001) and estimated VO2max (49.3 +/- 2.8 vs. 57.2 +/- 3.5 mg x kg LBW[-1] x min[-1], 0.05 < P < 0.10) were decreased in the relatives. The number of type IIb fibers (29.5 +/- 2.5 vs. 21.0 +/- 2.8%, P < 0.05) was increased in the relatives, whereas no significant differences were found in other fiber types or capillary density between the groups. Correlations were observed between number of type I fibers (positive), number of type IIb fibers (negative), and capillary density (positive) versus Rd as well as estimated VO2max (P < 0.05). In a multiple linear regression analysis with Rd as a dependent variable, estimated VO2max, family history of NIDDM, and number of type IIb fibers (P < 0.001, r2 = 0.64) significantly determined the level of Rd, whereas capillary density did not. In conclusion, insulin-resistant first-degree relatives of NIDDM patients are characterized by an increased number of type IIb muscle fibers. Whether this finding reflects a reduced physical activity level and fitness in the relatives or is of primary genetic origin remains to be determined. PMID- 9356033 TI - Clustering of long-term complications in families with diabetes in the diabetes control and complications trial. The Diabetes Control and Complications Trial Research Group. AB - We investigated familial clustering of diabetic retinopathy and nephropathy in the families of 372 subjects from the Diabetes Control and Complications Trial (DCCT). These subjects had 467 first-degree relatives with IDDM or NIDDM. Family sizes ranged from two to six. A complete data set was obtained from 241 relatives of 217 DCCT subjects. Among the DCCT subjects, 53% were in the intensive treatment group and 47% were in the conventional group; 44% were from the primary prevention cohort (no retinopathy or microalbuminuria at the DCCT baseline) and 56% were from the secondary intervention cohort (mild-to-moderate nonproliferative retinopathy and <200 mg/24 h albumin excretion rate [AER] at baseline). Retinopathy and nephropathy were assessed with seven-field stereo fundus photography and timed urinary AER measurements. Retinopathy was assessed using the Early Treatment Diabetic Retinopathy Study (ETDRS) scale. ETDRS scores and AERs were adjusted for the DCCT treatment group and for significant covariates from among sex, age, diabetes duration, HbA1c value, and body weight. Familial associations were assessed by comparing the prevalence of retinopathy and nephropathy in diabetic relatives of the respective positive versus negative DCCT subjects. To determine family clustering of the severity of retinopathy or nephropathy, the intraclass (familial) correlation was computed from the log adjusted retinopathy and nephropathy scores of DCCT subjects and their relatives for all family members and sib-sib relationships. For parent-offspring, mother child, and father-child relationships, the pairwise estimate of the correlations was computed. A correlation of 0.2 was considered to be biologically meaningful a priori. Among families of patients in the intensive and conventional groups combined, there was an increased risk of severe retinopathy (an ETDRS score > or =47, clinically significant macular edema, or laser treatment in either eye) among relatives of retinopathy-positive vs. retinopathy-negative DCCT subjects in the secondary intervention cohort (odds ratio [OR], 3.1; 95% CI 1.2-7.8; P < 0.05). There was no increase in the risk of retinopathy of any severity (microaneurysms or worse) in the relatives of retinopathy-positive vs. retinopathy-negative DCCT subjects of the primary prevention cohort. There was an increased risk of nephropathy (AER >40 mg/24 h) in relatives of nephropathy positive versus nephropathy-negative DCCT subjects of the secondary intervention cohort (OR, 5.4; 95% CI 2.2-13.7; P < 0.001). The risk of severe retinopathy in the relatives of positive versus negative subjects from the conventional treatment group alone (OR, 4.3; 95% CI 1.01-18.6; P < 0.05) was statistically significant and somewhat greater than that among relatives of the subjects in intensive treatment group (OR, 2.4; 95% CI 0.7-8.1), which was not significant. Correlations for the severity of retinopathy were 0.187 (all family members), 0.327 (parent-offspring), 0.249 (father-child), 0.391 (mother-child), and 0.060 (sib-sib), using the combined treatment group families. All these correlations were statistically significant (P < 0.05), except for sib-sib. The results showed similar trends when the families from the conventional and intensive treatment groups were analyzed separately. Correlations for nephropathy in the combined treatment group families were 0.063 (all family members), 0.138 (parent offspring), 0.170 (father-child), 0.103 (mother-child), and 0.107 (sib-sib). None of these correlations is statistically significant. The lack of significant correlation for the severity of nephropathy may reflect the relatively short duration of diabetes in the offspring of these families and the known high intrasubject variability of AERs. These data provide the first available evidence that the severity of diabetic retinopathy is influenced by familial (possibly genetic) factors and confirmatory evidence that such factors influence the development PMID- 9356034 TI - Impairment of cerebrovascular reactivity in long-term type 1 diabetes. AB - The early preclinical detection of cerebrovascular complications in individuals with diabetes is one of the goals of care described in the St. Vincent Declaration. In accordance with this goal, the aim of the present work was to investigate whether altered cerebral microvascular function in patients suffering from type 1 diabetes can be detected with a transcranial Doppler probe after the administration of acetazolamide. A total of 72 type 1 diabetic patients and 40 healthy control subjects entered the study. Patients were divided into two groups: those with long-term diabetes (disease duration of >10 years, n = 37) and those with short-term diabetes (disease duration of < or =10 years, n = 35). Mean blood-flow velocity in the middle cerebral artery (MCAV) was measured at rest and at 5, 10, 15, and 20 min after intravenous administration of 1 g acetazolamide with a transcranial Doppler probe and expressed as the percentage change from the pretest measurement. The percentage increase in MCAV (cerebrovascular reactivity) was calculated at each time point and compared between the groups. Cerebrovascular reserve capacity (CRC), expressed as the maximal percentage increase of the MCAV, was compared between the groups. Additionally, a reproducibility study of CRC was performed in 10 patients, using intraclass correlations. Cerebrovascular reactivity in the long-term diabetes group was lower (means +/- SD: 5 min, 23.4 +/- 15.4%; 10 min, 28.8 +/- 17.0%; 15 min, 30.0 +/- 15.6%; 20 min, 24.2 +/- 17.8%) than that of the control subjects (5 min, 43.5 +/- 23.9%; 10 min, 55.3 +/- 24.0%; 15 min, 56.7 +/- 23.8%; 20 min, 54.8 +/- 25.9%) and the short-term diabetic patients (5 min, 43.6 +/- 25.9%; 10 min, 52.2 +/- 27.7%; 15 min, 55.3 +/- 32.2%; 20 min, 45.8 +/- 35.8%). CRC was lower in the long-term diabetes group than in the control group or the short-term diabetes group. Impairment of cerebrovascular reactivity was associated with retino- and nephropathy and increased levels of fibrinogen. In contrast, CRC was independent from actual glucose, insulin, glycosylated hemoglobin, von Willebrand factor antigen, and alpha-2 macroglobulin levels. Transcranial Doppler measurements of the changes in MCAV after stimulation with acetazolamide can detect altered cerebral microvascular function in patients with diabetes. Cerebrovascular reactivity and reserve capacity are reduced in patients with long-term diabetes. Further prospective studies should delineate the clinical significance of our results. PMID- 9356035 TI - Aerobic exercise capacity remains normal despite impaired endothelial function in the micro- and macrocirculation of physically active IDDM patients. AB - The aim of the present study was to examine if diabetes in the absence of neuropathy affects the exercising capacity of IDDM patients, and whether regular, intense training has a beneficial effect on endothelial function. Five groups of subjects were studied: 23 healthy control subjects who exercised regularly (age 33 +/- 6 years), 23 nonneuropathic type 1 diabetic patients who exercised regularly (age 33 +/- 6 years, IDDM duration 11 +/- 8 years), 7 neuropathic type 1 diabetic patients who exercised regularly (age 36 +/- 7 years, IDDM duration 22 +/- 8 years), 18 healthy subjects who did not exercise regularly (age 34 +/- 7 years), and 5 nonneuropathic type 1 diabetic patients who did not exercise regularly (age 31 +/- 4 years, IDDM duration 20 +/- 3 years). All groups were matched for age, sex, and body weight. No differences existed in the energy expenditure per week in physical activity among the three exercising groups or between the two nonexercising groups. The maximal oxygen uptake was similar between control and diabetic nonneuropathic exercisers, and among diabetic neuropathic exercisers, control nonexercisers, and diabetic nonexercisers; however, a significant difference existed between the first two and the last three groups (P < 0.0001). A stepwise increase was found in the resting heart rate among the groups, ranging from the lowest in control exercisers to the highest in diabetic nonexercisers, but the maximal heart rate was lower only in diabetic neuropathic exercisers compared with all other groups (P < 0.05). Assessments of endothelial function in both macro- and microcirculation were performed in 12 control exercisers, 10 diabetic nonneuropathic exercisers, 5 diabetic neuropathic exercisers, 17 control nonexercisers, and 4 diabetic nonexercisers. When all diabetic patients were considered as one group and all control subjects as another, the microcirculation endothelial function in the diabetic group was reduced compared with the control subjects (91 +/- 49 vs. 122 +/- 41% flux increase over baseline; P < 0.05). In contrast, no differences existed among the three diabetic groups or between the two control groups. Similarly, in macrocirculation, a reduced response during reactive hyperemia was observed in the diabetic patients compared with control subjects (7.0 +/- 4.5 vs. 11.2 +/- 6.6% diameter increase; P < 0.05), whereas again no difference existed among the three diabetic groups or between the two control groups. These data suggest that diabetes per se does not affect aerobic exercise capacity (VO2max) in physically active individuals, but is reduced in the presence of neuropathy. In addition, regular exercise training involving the lower extremities does not improve the endothelial function in the micro- and macrocirculation of the nonexercised upper extremity in type 1 diabetic patients. PMID- 9356036 TI - Defective plasma antioxidant defenses and enhanced susceptibility to lipid peroxidation in uncomplicated IDDM. AB - Oxidative stress is postulated to be increased in patients with IDDM. Accumulating evidence suggests that oxidative cell injury caused by free radicals contributes to the development of IDDM complications. On the other side, a decreased efficiency of antioxidant defenses (both enzymatic and nonenzymatic) seems to correlate with the severity of pathological tissue changes in IDDM. Thus, we determined plasma antioxidant defenses, measuring the total radical trapping antioxidant capacity (TRAP) and the two markers of oxidative stress, lipid hydroperoxides (ROOHs) and conjugated dienes, in 72 patients with well controlled IDDM and without evident complications, compared with 45 nondiabetic subjects. Compared with control subjects, IDDM patients showed significantly reduced plasma TRAP (669 +/- 131 vs. 955 +/- 104 micromol/l, P < 0.001) and significantly increased levels of ROOHs (7.13 +/- 2.11 vs. 2.10 +/- 0.71 micromol/l, P < 0.001) and conjugated dienes (0.0368 +/- 0.0027 vs. 0.0328 +/- 0.0023 arbitrary units [AU], P < 0.01), especially in the trans-trans conformation (0.0340 +/- 0.0028 vs. 0.0259 +/- 0.0022 AU, P < 0.001), with a concurrent reduction of conjugated dienes in the cis-trans conformation (0.0028 +/- 0.0011 vs. 0.0069 +/- 0.0012 AU, P < 0.001). The oxidative parameters studied did not appear to be correlated with metabolic control (HbA1c levels) and lipid profile (cholesterol or triglyceride levels). The reduced TRAP and the increased ROOH and conjugated diene plasma levels, together with the decreased ratio of cis trans/trans-trans conjugated dienes, which reflects an altered redox status of plasma, indicate that in IDDM patients, oxidative stress is enhanced and antioxidant defenses are defective, regardless of diabetes duration, metabolic control, or presence of complications. PMID- 9356037 TI - The blood contribution to early myocardial reperfusion injury is amplified in diabetes. AB - Cardiovascular disease is excessive in diabetes, and blood cell function is altered. It is not clear, however, if alterations in the blood contribute to the excessive cardiovascular complications of this disease. In this study, we compared the contribution of nondiabetic and diabetic blood to myocardial reperfusion injury. The recovery of cardiac contractile function following no flow ischemia was studied in isolated diabetic and nondiabetic rat hearts perfused with diabetic or nondiabetic diluted whole blood. Hearts were isolated from 10- to 12-week-old diabetic (streptozotocin, 65 mg/kg, i.v.) and nondiabetic rats and perfused with a Krebs-albumin-red cell solution (K2RBC, Hct 20%). After a 30-min pre-ischemic control period, during which cardiac pump function was evaluated, diabetic and nondiabetic hearts were perfused for 5 min with diluted whole blood (DWB; Hct 20%) collected from either diabetic or nondiabetic donor animals. Coronary flow was then stopped and the hearts subjected to 30 min of no flow ischemia. Following ischemia, the hearts were reperfused with the K2RBC perfusate. Cardiac contractile function was evaluated throughout the 60-min reperfusion period. Six groups were studied: diabetic and nondiabetic hearts perfused before ischemia with either K2RBC, nondiabetic DWB (NDDWB), or diabetic DWB (DDWB). Perfusion with DWB prior to ischemia impaired the recovery of contractile function in all cases. The impairment to recovery was greater with DDWB than with NDDWB. Although diabetic hearts perfused with K2RBC throughout recovered quite well, the effect of DDWB perfusion in the diabetic hearts was dramatic. In an effort to determine why diabetic blood impaired functional recovery, measures of blood filterability and the generation of reactive oxygen species (ROS) were made. We found that diabetic blood was less filterable than nondiabetic blood; that is, the diabetic blood cells tended to plug the 5-microm filter pores more readily than the nondiabetic blood cells. Also, we found that the diabetic blood was capable of generating significantly greater ROS (oxygen free radicals) than nondiabetic blood (P < 0.05). These findings suggest that the blood contribution to myocardial reperfusion injury is amplified in diabetes. A tendency for diabetic blood cells to plug capillary-sized pores and show enhanced oxygen free radical production may account for the excessive contribution of diabetic blood to reperfusion injury in the heart. PMID- 9356038 TI - Renal and ocular hemodynamic effects of insulin. AB - There is evidence that the vasodilator action of insulin is mediated by the release of nitric oxide (NO). We hypothesized that euglycemic hyperinsulinemia might increase renal and ocular blood flow, and that the vasodilator capacity of insulin might be NO-dependent. Euglycemic insulin clamps were performed in 10 healthy subjects. Sixty minutes after the start of insulin administration, an intravenous coinfusion of N-monomethyl-L-arginine (L-NMMA), an inhibitor of NO synthase, or of norepinephrine (NE), an endothelium-independent vasoconstrictor, was started. Renal plasma flow was measured by para-aminohippurate (PAH) clearance method. Ocular hemodynamics were assessed by laser interferometric measurement of fundus pulsations and Doppler sonographic measurement of blood flow velocity in the ophthalmic artery. Renal plasma flow and ocular fundus pulsations were increased by insulin. L-NMMA almost completely abolished the vasodilative effects of insulin, whereas the effects of combined infusion of insulin and NE were approximately the sum of the hemodynamic changes induced by each agent alone. The results show that during euglycemic hyperinsulinemia, renal and ocular blood flow are increased, which may be mediated either by a local vasodilator effect or a systemic increase in flow. The hemodynamic effects of insulin in the kidney and the eye are at least partially dependent on NO synthesis. Because the insulin plasma levels we obtained are in the high physiological range, it may be assumed that insulin plays a role in renal and ocular blood flow regulation. PMID- 9356039 TI - Immunohistochemical quantification of heparan sulfate proteoglycan and collagen IV in skeletal muscle capillary basement membranes of patients with diabetic nephropathy. AB - In IDDM patients, an increased permeability of the glomerular capillaries has been associated with a general loss of negatively charged heparan sulfate proteoglycans (HSPGs) within basement membranes (BMs). In the present study, we used immunohistochemical staining to quantify heparan sulfate (HS), HSPG core protein, and collagen IV in capillary basement membranes of skeletal muscle biopsies taken from 9 healthy control subjects (C) and 20 IDDM patients: 7 with normal albumin excretion rate (<30 mg/24 h) (D0), 5 with incipient nephropathy (albumin excretion rate 30-300 mg/24 h) (D1), and 8 with clinical nephropathy (albumin excretion rate >300 mg/24 h) (D2). In the capillaries, staining was measured by a scanning and integrating microspectrophotometer. A significant difference in the absorbance of HS was found among the four subgroups (means +/- SD): 0.477 +/- 0.082 (C), 0.627 +/- 0.031 (D0), 0.542 +/- 0.098 (D1), and 0.371 +/- 0.118 (D2) (P = 0.006). Similarly, an overall significant difference in the absorbance of collagen IV was demonstrated (means +/- SD): 0.836 +/- 0.111 (C), 0.838 +/- 0.300 (D0), 0.970 +/- 0.173 (D1), and 0.512 +/- 0.248 (D2) (P = 0.02). No statistical difference in the absorbance of core protein was demonstrated among the groups. Within the diabetic groups, HS was inversely correlated to albuminuria (r = -0.76, P = 0.003) and albuminuria corrected for creatinine clearance (r = -0.69, P = 0.008). Because, in IDDM patients with albuminuria, alterations of the content of HS and collagen IV within the capillary BM have been demonstrated immunohistochemically, not only in the glomerular filtration barrier, but also in the skeletal muscle capillary BM, we suggest that these changes reflect universal quantitative or qualitative alterations within the capillary filtration barrier. PMID- 9356041 TI - The effect of HLA-B allele on the IDDM risk defined by DRB1*04 subtypes and DQB1*0302. AB - The genes encoding the HLA-DQ heterodimer molecules, DQB1 and DQA1, have been found to have the strongest association with IDDM risk, although there is cumulative evidence for the effect of other gene loci within the major histocompatibility complex gene region. After the HLA-DQ locus, the HLA-DR locus has been suggested most often as contributing to the disease susceptibility. In this study we analyzed at the population level the effect of DR4 subtypes and class I, HLA-B alleles, on IDDM risk when the influence of the DQ locus was stratified. In all three populations studied (Estonian, Latvian, and Russian), DQB1*0302 haplotypes most frequently carried DRB1*0401 or DRB1*0404. DRB1*0401 was the most prevalent subtype in IDDM patients, whereas DRB1*0404 was decreased in frequency. DRB1*0402 was also prevalent among Russian haplotypes, but was not associated with IDDM risk. When HLA-B alleles were analyzed, strong associations between the presence of specific B alleles and DRB1*04 subtypes were detected. The HLA-B39 allele was found significantly more often in DRB1*0404-DQB1*0302 positive patients than in healthy control subjects positive for this haplotype: 27 of 54 (50%) vs. 4 of 49 (8.2%) (P < 0.0001). The results demonstrate that DQ and DR genes cannot explain all of the HLA-linked susceptibility to IDDM, and that the existence of a susceptibility locus telomeric to DR is probable. PMID- 9356040 TI - Upregulation of mesangial growth factor and extracellular matrix synthesis by advanced glycation end products via a receptor-mediated mechanism. AB - Enhanced advanced glycosylation end product (AGE) formation has been shown to participate in the pathogenesis of diabetes-induced glomerular injury by mediating the increased extracellular matrix (ECM) deposition and altered cell growth and turnover leading to mesangial expansion. These effects could be exerted via an AGE-receptor-mediated upregulation of growth factors, such as the IGFs and transforming growth factor-beta (TGF-beta). We tested this hypothesis in human and rat mesangial cells grown on nonglycated or native bovine serum albumin (BSA), glycated BSA with AGE formation (BSA-AGE), or glycated BSA in which AGE formation was prevented by the use of aminoguanidine (BSA-AM), in the presence or absence of an antibody, alpha-p60, directed against the p60/OST protein named AGE receptor 1 (AGE-R1), or normal control (pre-immune) serum. The mRNA and/or protein levels of IGF-I, IGF-II, IGF receptors, IGF binding proteins (IGFBPs), TGF-beta1 and the ECM components fibronectin, laminin, and collagen IV were measured, together with cell proliferation. Both human and rat mesangial cells grown on BSA-AGE showed increased IGF-I and total and bioactive TGF-beta medium levels and enhanced IGF-I, IGF-II, and TGF-beta1 gene expression, compared with cells grown on BSA, whereas total IGFBP and IGFBP-3 medium content, IGF receptor density and affinity, and IGF-I receptor transcripts were unchanged. Moreover, cells grown on BSA-AGE showed increased ECM protein and mRNA levels versus cells cultured on BSA, whereas cell proliferation was unchanged in human mesangial cells and slightly reduced in rat mesangial cells. Growing cells on BSA-AM did not affect any of the measured parameters. Co-incubation of BSA-AGE with anti-AGE R1, but not with pre-immune serum, prevented AGE-induced increases in IGF-I, TGF beta1, and ECM production or gene expression; anti-AGE-R1 also reduced growth factor and matrix synthesis in cells grown on BSA. These results demonstrate that mesangial IGF and TGF-beta1 synthesis is upregulated by AGE-modified proteins through an AGE-receptor-mediated mechanism. The parallelism with increased ECM production raises the speculation that the enhanced synthesis of these growth factors resulting from advanced nonenzymatic glycation participates in the pathogenesis of hyperglycemia-induced mesangial expansion. PMID- 9356042 TI - Association of HLA-DR, DQ genotype with different beta-cell functions at IDDM diagnosis in Japanese children. AB - Japanese IDDM patients have been demonstrated to have unique and different HLA associations from white patients. To elucidate the effect of HLA-associated genetic factors on the clinical heterogeneity of IDDM in Japanese people, HLA DRB1, DQA1, and DQB1 genotypes in 88 childhood-onset Japanese IDDM patients were examined by polymerase chain reaction-sequence-specific oligonucleotide (PCR-SSO) or sequence-specific primers (SSP). Of the 88 IDDM patients, 26 (29.5%) had DRB1*0405-DQA1*0302-DQB1*0401/X (DR4-DQ4/X), 38 (43.2%) had DRB1*0901-DQA1*0302 DQB1*0303/X (DR9-DQ9/X), and 9 (10.2%) were DR4/9-DQ4/9 heterozygous in the present study (X does not contain protective alleles). Clinical heterogeneity such as age distribution at onset, prevalence and serum level of anti-GAD antibodies (GADAb), and residual pancreatic beta-cell function after diagnosis were compared between patients with HLA-DR4-DQ4 and DR9-DQ9. The frequency of DR9 DQ9 genotype was significantly higher in the younger (0-10 years) than in the older (11-16 years) age-group of onset, but the frequency of DR4-DQ4 was higher in the older (11-16 years) age-group. Although no association of DR-DQ genotypes with the prevalence and serum level of GADAb was found among newly diagnosed patients, long-standing DR9-DQ9 patients had significantly higher levels of GADAb than those with DR4-DQ4. While no difference in time course of serum C-peptide (CPR) levels was detected between GADAb+ and GADAb- patients, a remarkable difference was demonstrated between DR9-DQ9 and DR4-DQ4 patients. The residual pancreatic beta-cell function was retained more in patients with DR4-DQ4 than in those with DR9-DQ9 at diagnosis through 12-18 months after diagnosis. These results suggest that the DR9-DQ9 genotype may induce stronger autoimmune destructive response (T-helper 1 function) against target beta-cells than the DR4 DQ4 genotype does. Our findings may warrant further studies on the association of diabetogenic autoimmune response with HLA class II molecules and contribute to a clarification of interracial differences in HLA-encoded susceptibility to IDDM. PMID- 9356043 TI - The Gln223Arg and Lys656Asn polymorphisms in the human leptin receptor do not associate with traits related to obesity. PMID- 9356045 TI - No coding mutations are detected in the peroxisome proliferator-activated receptor-gamma gene in Japanese patients with lipoatrophic diabetes. PMID- 9356044 TI - Dominant transmission of insulin resistance in a type A family resulting from a heterozygous nonsense mutation in the insulin receptor gene and associated with decreased mRNA level and insulin binding sites. PMID- 9356046 TI - Progressive islet graft failure occurs significantly earlier in autoantibody positive than in autoantibody-negative IDDM recipients of intrahepatic islet allografts. AB - Alloimmunity has been uncovered to be a cause of graft loss representing a major barrier for clinical islet transplantation, and several studies are designed to evaluate new strategies for immunosuppression to prevent alloimmunity. In contrast, the significance for autoimmune destruction of transplanted beta-cells has remained somewhat controversial. Recently, two case reports based on histological findings have suggested recurrent autoimmune insulitis despite immunosuppressive therapy both in clinical pancreas and in islet transplantation. In the present study, in 23 islet-grafted patients with IDDM receiving standard immunosuppressive therapy, we demonstrate that progressive impairment of islet graft function occurs significantly earlier in those individuals positive for autoantibodies as a typical stigma of diabetes-associated autoimmunity that is well established in the prediabetic periods of IDDM. Intraportal infusion of allogeneic islets was performed in 23 C-peptide-negative IDDM patients, according to the clinical transplantation categories defined as islet after kidney (IAK) or simultaneous islet and kidney (SIK). Complete islet graft failure was defined as the 1st day of permanent C-peptide negativity in the serum (<0.2 ng/ml) and C peptide negativity in the urine (<2 microg/dl). The median observation period following islet transplantation was 12 months (range 1-50) with a cumulative follow-up of 336 months. Islet cell antibodies (ICAs) and GAD65 antibodies were monitored before and regularly after islet transplantation. Kaplan-Meier survival analysis and log-rank statistics revealed a significant (P < 0.05) difference in cumulative islet graft survival depending on the presence of islet cell and/or GAD65 antibodies. These results strongly suggest that recurrent autoimmunity directed to transplanted beta-cells contributes to islet graft failure despite sustained immunosuppression. For successful clinical islet transplantation in the future, new immunosuppressive therapies are needed to prevent both alloimmunity and autoimmunity. PMID- 9356047 TI - Central leptin stimulates corticosterone secretion at the onset of the dark phase. AB - Leptin, a hormone secreted by adipose tissue in proportion to body adiposity, is proposed to be involved in the central nervous regulation of food intake and body weight. In addition, evidence is emerging that leptin regulates neuroendocrine and metabolic functions as well, presumably via its action in the central nervous system (CNS). To investigate this regulatory effect of leptin, we infused 3.5 microg of human leptin directly into the third cerebral ventricle (i3vt) of lean male Long-Evans rats, 90 min before the onset of their dark phase. Before and after infusion, blood samples were withdrawn through indwelling catheters for assessment of hormonal (plasma corticosterone, insulin, leptin), autonomic (plasma norepinephrine, epinephrine), and metabolic (plasma glucose) parameters. I3vt leptin caused an increase in plasma corticosterone and plasma leptin levels relative to the control condition. The effects of i3vt leptin on corticosterone secretion became particularly apparent after the onset of the dark phase. The results of the present study indicate that i3vt leptin stimulates the hypothalamo pituitary-adrenal (HPA) axis, particularly when rats normally encounter their largest meals. These results are consistent with the possibility that high circulating leptin levels may underlie the increased activity of the HPA axis that is generally characteristic of human obesity and most animal models of obesity. PMID- 9356048 TI - Nitric oxide stimulates skeletal muscle glucose transport through a calcium/contraction- and phosphatidylinositol-3-kinase-independent pathway. AB - Recently published data have provided evidence that nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) are signaling intermediates in the pathway through which muscle contraction stimulates glucose transport. As exercise promotes both NO production and calcium flux, we examined the relationships between NO stimulated glucose uptake and calcium-, contraction-, and phosphatidylinositol-3 kinase (PI-3-K)-mediated glucose transport in the isolated incubated rat epitrochlearis muscle preparation. The NO donor sodium nitroprusside (SNP; 10 mmol/l) and dibutyryl cGMP (100 micromol/l) accelerated epitrochlearis glucose transport four- to fivefold above basal levels (P < 0.001) in a manner similar to in vitro contractile activity and the calcium releasing agent N-(6-aminohexyl)-5 chloro-1-naphthalenesulfonamide (W7; 100 micromol/l). In the case of SNP, this effect could be completely attributed to an increase in cell surface GLUT4. The effect of SNP on glucose transport was not inhibitable by either wortmannin (1.5 micromol/l) or dantrolene (12.5 micromol/l). Similarly, neither calcium nor contraction stimulation of glucose transport was affected by the NO synthase inhibitors NG-monomethyl-L-arginine (L-NMMA; 100 micromol/l) or 7-nitroindazole (1 mmol/l). Furthermore, whereas SNP raised epitrochlearis cGMP levels tenfold (P < 0.001), neither in vitro contractile activity nor W7 significantly elevated cGMP. These results indicate that NO/cGMP can markedly stimulate skeletal muscle glucose transport by increasing GLUT4 levels at the cell surface by a mechanism that does not depend on activation of PI-3-K. In addition, since calcium/contraction-stimulated glucose transport is not blocked by NO synthase inhibition and did not elevate cGMP, NO/cGMP may be part of a novel pathway that is distinct from both the insulin- and contraction-activated mechanisms. PMID- 9356049 TI - Physician resource utilization after geriatric trauma. AB - OBJECTIVE: As health care resources become increasingly strained, the value of physician consultation has come under heightened scrutiny. This report reviews the value of early consultation by the physical medicine and rehabilitation (PMR) service to an integrated trauma service for geriatric patients with multiple trauma. METHODS: We retrospectively reviewed the records of 110 geriatric trauma patients (age > 60 years) with an Injury Severity Score > or = 15 to evaluate the effects of PMR consultation. Patients in group 1 were admitted to a general surgical service, and those in group 2 were admitted to a multidisciplinary trauma service. Demographic and physiologic factors, as well as short-term and long-term outcomes, were evaluated, and a subgroup analysis was performed to compare early (< or =3 days) versus late (>3 days) consultation by PMR. RESULTS: Although there were significant differences in Glasgow Coma Scale score and length of stay, no differences were found within groups in other demographic, physiologic, or outcome data. Focused review of PMR intervention based on early versus late consultation revealed no significant difference between the two groups. Furthermore, an after-discharge phone survey revealed no significant group differences in dependence on a care provider or nursing home placement, readmission to hospital, employment status, or current functional activity status. CONCLUSIONS: Long-term patient functional outcome and the in-house rehabilitation process are not affected by integration of PMR into a multidisciplinary trauma team or early PMR consultation. PMID- 9356050 TI - Comorbid conditions in old patients with femur fractures. AB - BACKGROUND AND METHODS: The incidence of preexisting medical diseases (comorbid conditions) and their influence on the high rate of falls, associated severe injuries, operative treatment, and outcome including mortality rate, duration of hospitalization, and rehabilitation success was retrospectively evaluated in a group of 102 patients (mean age, 81 years; 81% women) with femoral fractures. A comparison of polymorbidity rates in a control group of 102 patients (mean age, 79 years; 86% women) with proximal humeral fractures was added. RESULTS: The associated polymorbidity rate among patients with femoral fractures (FF) usually was statistically significantly higher than among patients with proximal humeral fractures (PHF) despite a comparable age and sex distribution: 80% of the patients with FF presented with cardiovascular (p < or = 0.001), 41% with pulmonary (p < 0.001), 67% with gastrointestinal (p < or = 0.001), 71% with neurologic (p < or = 0.001), 55% with urologic (p < or = 0.001), 75% with musculoskeletal (p < or = 0.1), and 61% with psychiatric (p < or = 0.001) disorders and complaints. Ninety percent of the patients used different medications (diuretics, cardiac agents, anticoagulants, antidiabetic agents, steroids, hypnotics, analgesics, psychotropic agents). The postoperative mortality rate was 11%, and the mean hospitalization period was 30 days. Forty nine percent of the patients were discharged to their homes. Only 56% of the patients with PHF, however, presented with cardiovascular, 8% with pulmonary, 11% with gastrointestinal, 8% with neurologic, 9% with urologic, 64% with musculoskeletal, and 10% with psychiatric disorders. CONCLUSION: The polymorbidity in the old patient probably is a major intrinsic cause of the high incidence of falls and associated severe femoral fractures. It influences the perioperative and postoperative medical and anesthesiologic treatment, the postoperative mortality rate, and the duration and success of the postoperative rehabilitation phase. PMID- 9356051 TI - Open reduction and internal fixation of the distal humerus: functional outcome in the elderly. AB - OBJECTIVE: To examine the functional outcome of a cohort of elderly patients after open reduction and internal plate and screw fixation of distal humerus fractures. DESIGN: Retrospective review of a consecutive series of patients older than 60 years of age who underwent plate and screw fixation of a distal humerus fracture. MATERIALS AND METHODS: Eighteen patients, aged 63 to 85 years (average, 71 years), underwent open reduction and internal fixation of a displaced distal humerus fracture using plates and screws. Three were Arbeitsgemeinschaft fur Osteosynthesefragen/Association for the Study of Internal Fixation (AO/ASIF) type A, 2 were type B, and 13 were type C fractures. The patients were reviewed at a minimum follow-up of 1 year after surgery. MEASUREMENTS AND MAIN RESULTS: All patients had a good or excellent clinical result using a standardized method of evaluation. General health status, as measured by the SF-36 Health Survey, was comparable to the published norms for U.S. male and female populations of similar age. CONCLUSION: Open reduction and internal fixation of the distal humerus in the elderly can provide good clinical results. Good clinical results, however, do not imply good general health status. PMID- 9356052 TI - Probability model of hospital death for severe trauma patients based on the Simplified Acute Physiology Score I: development and validation. Archivio Diagnostico. AB - BACKGROUND: We evaluated whether or not the Simplified Acute Physiology Score (SAPS) I is a suitable audit system for trauma patients admitted to general intensive care units (ICUs). A probability model for SAPS I was retrospectively assessed on trauma patients admitted to general ICUs from 1990 to 1992. Because it was determined that SAPS did not fit the data well, we developed a customized probability model of SAPS I for trauma patients and validated it prospectively on an independent data set (patients admitted to general ICU in 1993-1994). Measures of calibration (goodness of fit) and discrimination (receiver operating characteristic curve) were adopted to assess the performance of the model. METHODS: A multicenter study was performed involving general ICUs in northern Italy that participated in a project of standardized data collection. This project is known in Italy under the acronym of ARCHIDIA (Diagnostic Archive). All patients consecutively admitted to the ICU were registered in the data base from January 1, 1990, to December 31, 1994 (16,767 patients). For the SAPS analysis, we considered only patients who fulfilled the following criteria: older than 17 years, length of stay longer than 24 hours, and SAPS I correctly computed. Inclusion criteria resulted in 12,156 patients analyzed; there were 1,936 patients with trauma. RESULTS: Trauma patients represented more than 15% of all patients admitted to ICUs during the period considered. Compared with the general population, they were younger (41.2 vs. 56.7 years), stayed in ICUs for longer periods (12.4 vs. 9.5 days), and had a lower hospital mortality rate (17.8 vs. 28.7%). Because the original probability model did not perform well among patients with trauma, a new model was developed and prospectively validated only for trauma patients (customization). Measures of calibration and discrimination showed a good performance both in the developmental (goodness of fit: chi2 = 8.47; p = 0.38; area under the curve = 0.94 +/- 0.01) and in the validation (goodness of fit: chi2 = 12.75; p = 0.12; area under the curve = 0.85 +/- 0.01) data sets. CONCLUSION: Customization of SAPS I for trauma patients has shown good calibration and high discriminatory power in Italian ICUs and when applied to an independent data base. The advantage of customization would be the collection of the same set of variables for all patients admitted to ICUs against the use of specific scoring systems. PMID- 9356053 TI - Lessons learned: durability and progress of a program for ancillary cost reduction in surgical critical care. AB - OBJECTIVE: Modern surgical care must meet high standards of quality but must also be cost-effective. Critical care uses huge amounts of resources, and strategies for effective use of scarce, expensive intensive care unit beds must be implemented. Previously, we demonstrated that ancillary expenditures can be decreased without compromising care. The present study was performed to determine whether our cost-containment strategies were durable and could be extended to areas, such as chest roentgenography, where savings previously proved elusive. METHODS: Costs for laboratory tests, radiographs, and drugs were determined prospectively for all surgical intensive care unit care for a 34-month period (January 1, 1994-October 31, 1996) at an urban university center. A systematic, multidisciplinary cost-reduction program began on May 1, 1994, with emphasis on laboratory and radiographic testing and procedures and drug therapies. Calendar year cohorts were compared by age and Acute Physiology and Chronic Health Evaluation II and III admission scores. Outcome variables were hospital mortality, days in the intensive care unit and hospital, and expenditures. Cost data were taken weekly from the hospital's clinical information system. RESULTS: All admission noncost variables were identical. There were significant reductions in intensive care unit and hospital length of stay, and there was a trend (p = 0.07) toward decreased hospital mortality. Normalized by the number of patient days per week, arterial blood gas determinations were reduced 46% between 1994 and 1996, and nonarterial blood gas laboratory tests were reduced by 29% (both p < 0.0001). Within the latter group, electrolyte determinations decreased by 38% and serum creatinine determinations decreased by 32%. Chest roentgenograms decreased by 34%, but pharmaceutical costs decreased by a remarkable 73%. CONCLUSION: Durable reductions in physician-ordered ancillary expenditures are possible without compromising the standard of care of critically ill patients, but active management and daily reinforcement are necessary to the process. Shorter length of stay and lower costs benefit the patient, the surgeon, the intensivist, and the institution. PMID- 9356054 TI - Outcome and cost analysis for outpatient skin grafting. AB - BACKGROUND: To reduce cost, outpatient surgery is advocated when feasible; however, the potential of compromising outcome is a concern. The purpose of this review is to assess patient outcome and cost for managing operative burn injuries without hospitalization. METHODS: During the past 18 months, 54 patients were identified with burns amenable to operative debridement and skin grafting without hospitalization. Twenty patients chose to be hospitalized and underwent prompt skin grafting. Operative skin grafting as an outpatient was chosen by the remaining 34 patients. Of these, four patients were subsequently hospitalized postoperatively (two for pain, one for cellulitis, and one for vomiting). RESULTS: Hospitalized patients and outpatients were similar in age and extent of burn; however, those hospitalized underwent skin grafting sooner after injury (2.1 +/- 0.4 days for inpatients vs. 11.5 +/- 0.8 days for outpatients; mean +/- SEM). Inpatients also had a significantly larger area skin-grafted (286 +/- 24 cm2 for inpatients vs. 178 +/- 14 cm2 for outpatients). Graft take was very good in each group. Cost, as indexed by patient charge, was substantially less for outpatients ($2,397 +/- $222) than for inpatients ($17,220 +/- $410). CONCLUSION: These results demonstrate a significant cost reduction with nonhospitalized operative care of burn injuries without any overt detriment in outcome, thus endorsing outpatient skin grafting when amenable. This review also illustrates that delaying operative intervention reduces the burn area required for grafting. PMID- 9356055 TI - Effects of burns on inhalation injury. AB - BACKGROUND: There are few studies of smoke injury combined with thermal burn. METHODS: Seven sheep (G1) received smoke injury alone; eight (G2) received a 40% full-thickness scald burn immediately after smoke injury. All animals were resuscitated with lactated Ringer's solution and killed 48 hours after injury. Cardiopulmonary variables and blood gases were measured serially. Ventilation perfusion distribution was analyzed using the multiple inert gas elimination technique. Lung wet to dry weight ratio and malondialdehyde levels were determined. RESULTS: G2 resulted in early significant hemodynamic changes. Serum total protein concentration was significantly lower and malondialdehyde significantly higher in G2. However, PaO2, lung wet to dry weight ratio, and ventilation perfusion mismatching in G2 did not differ from those in G1. CONCLUSIONS: Although the addition of burn injury exaggerated the lung lipid peroxidation and hypoproteinemia in the presence of more pronounced hemodynamic changes, the pulmonary dysfunction was not accentuated. PMID- 9356056 TI - Ineffectiveness of on-site intravenous lines: is prehospital time the culprit? AB - The purpose of the present study was to test the association between on-site intravenous fluid replacement and mortality in patients with severe trauma. The effect of prehospital time on this association was also evaluated. The design was that of an observational quasi-experimental study comparing 217 patients who had on-site intravenous fluid replacement (IV group) with an equal number of matched patients for whom this intervention was not performed (no-IV group). The patients were individually matched on their Prehospital Index obtained at the scene and were included in the study if they had an on-site Prehospital Index score > 3 and were transported alive to the hospital. The outcome measure of interest was mortality because of injury. The patients in the IV group had a significantly lower mean age (37 vs. 45 years; p < 0.001) and higher incidence of injuries to the head or neck (46 vs. 32%; p = 0.004), chest (34 vs. 17%; p < 0.001), and abdomen (28 vs. 12%; p < 0.001). The IV group also had a higher proportion of patients injured by motor vehicle crashes (41 vs. 27%; p = 0.003), firearms (9 vs. 2%; p = 0.001), and stabbing (20 vs. 9%; p = 0.001). The rate of extremity injuries (38 vs. 59%; p < 0.001) and falls (12 vs. 40%; p < 0.001) was lower for the IV group. In addition, the mean Injury Severity Score was significantly higher for the IV group (15 vs. 9; p < 0.001). The mortality rates for the IV and no-IV groups were 23 and 6% (p < 0.001). Logistic regression analysis showed that after adjusting for patient age, gender, Injury Severity Score, mechanism of injury, and prehospital time, the use of on-site intravenous fluid replacement was associated with a significant increase in the risk of mortality (adjusted odds ratio = 2.3; 95% confidence interval = 1.02-5.28; p = 0.04). To further evaluate the effect of prehospital time on the association between on-site IV use and mortality, the analysis was repeated separately for the following time strata: 0 to 30 minutes, 31 to 60 minutes, and >60 minutes. The adjusted odds ratios (95% confidence interval) for these strata were 1.05 (0.08-14.53; p = 0.97), 3.38 (0.84-13.62; p = 0.08), and 8.40 (1.27-54.69; p = 0.03). These results show that for prehospital times of less than 30 minutes, the use of on site intravenous fluid replacement provides no benefit, and that for longer times, this intervention is associated with significant increases in the risk of mortality. The results of this observational study have shown that the use of on site intravenous fluid replacement is associated with an increase in mortality risk and that this association is exacerbated by, but is not solely the result of, increased prehospital times. Our findings are consistent with the hypothesis that early intravenous fluid replacement is harmful because it disrupts the normal physiologic response to severe bleeding. Although this evidence is against the implementation of on-site intravenous fluid replacement for severely injured patients, further studies including randomized controlled trials are required to provide a definitive answer to this question. PMID- 9356057 TI - Solid viscus injury predicts major hollow viscus injury in blunt abdominal trauma. AB - BACKGROUND: As nonoperative management of blunt abdominal trauma has become more popular, reliable models for predicting the likelihood of concomitant hollow viscus injury in the hemodynamically stable patient with a solid viscus injury are increasingly important. METHODS: The Pennsylvania Trauma Systems Foundation registry was reviewed for the period from January 1992 to December 1995 for all adult (age > 12 years) patients with blunt trauma and an Abbreviated Injury Scale (AIS) score > or = 2 for a solid viscus (kidney, liver, pancreas, spleen). Patients with an initial systolic blood pressure < 90 mm Hg were excluded. Hollow viscus injuries included only lacerations or perforations of the gallbladder, gastrointestinal tract, or urinary tract. RESULTS: In the 4-year period, 3,089 patients sustained solid viscus injuries, 296 of whom had a hollow viscus injury (9.6%). The mean age was 35.6 years, mean Injury Severity Score was 22.2, and mean Revised Trauma Score was 7.3; 63.3% of the patients were male. A solitary solid viscus injury occurred in 2,437 patients (79%), 177 of whom (7.3%) had a hollow viscus injury. The frequency of hollow viscus injury increased with the number of solid organs injured: 15.4% of patients with two solid viscus injuries (n = 547) and 34.4% of patients with three solid viscus injuries (n = 96) suffered a concomitant hollow viscus injury (p < 0.001 vs. one organ). A hollow viscus injury was 2.3 times more likely for two solid viscus injuries and 6.7 times more likely for three solid viscus injuries compared with a solitary solid viscus injury. For solitary solid viscus injury, the frequency of hollow viscus injury varied little with increasing AIS score (AIS score 2, 6.6%; AIS score 3, 8.2%; AIS score 4, 9.2%; AIS score 5, 6.2%) (p = 0.27 between groups), suggesting that the incidence of hollow viscus injury is related more to the number of solid visceral injuries than the severity of individual organ injury. Also, when the sum of the AIS scores for solid viscus injuries was <6, the mean rate of hollow viscus injury was 7.8%. This increased to 22.8% when the sum of the AIS scores for solid viscus injury was > or =6 (p < 0.001). A pancreatic injury in combination with any other solid viscus injury had a rate of hollow viscus injury of >33%. CONCLUSION: A model of organ injury scaling predicted hollow viscus injury. Multiple solid viscus injuries, particularly pancreatic, or abdominal solid viscus injuries with an AIS score > or = 6, were predictive of hollow viscus injury. Identification of these injury patterns should prompt consideration for early operative intervention. PMID- 9356058 TI - Penetrating left thoracoabdominal trauma: the incidence and clinical presentation of diaphragm injuries. AB - OBJECTIVE: The objective of this study was to (1) determine the incidence of diaphragmatic injuries in penetrating left thoracoabdominal trauma and (2) evaluate the role of laparoscopy in detecting clinically occult diaphragmatic injuries. PATIENTS AND METHODS: One hundred nineteen consecutive patients with penetrating injuries to the left thoracoabdominal region presenting to Los Angeles County-University of Southern California Medical Center were prospectively evaluated during an 8-month period. Either celiotomy (with hemodynamic instability or peritonitis) or laparoscopy was performed. Results of the clinical examination and roentgenographic findings were recorded preoperatively. RESULTS: One hundred seven patients were fully evaluated. Fifty patients required emergent celiotomy. Fifty-seven patients underwent laparoscopy. The overall incidence of diaphragmatic injuries was 42% (59% for gunshot wounds, 32% for stab wounds). Among the 45 patients with diaphragmatic injuries, 31% had no abdominal tenderness, 40% had a normal chest roentgenogram, and 49% had an associated hemopneumothorax. Fifteen of the patients undergoing laparoscopy (26%) had occult diaphragm injuries. CONCLUSION: (1) The incidence of diaphragmatic injuries in association with penetrating left thoracoabdominal trauma is high. (2) The clinical and roentgenographic findings are unreliable at detecting occult diaphragmatic injuries. (3) Laparoscopy is a vital tool for detecting occult diaphragmatic injuries among patients who have no other indications for formal celiotomy. PMID- 9356059 TI - Direct induction of acute lung and myocardial dysfunction by liver ischemia and reperfusion. AB - OBJECTIVES: To investigate whether liver ischemia and reperfusion (IR) directly affect functions of remote organs. BACKGROUND: Cardiovascular and respiratory dysfunction follows hemorrhage, spinal shock, or trauma as a result of no-flow reflow phenomena. Hepatic IR induces remote organ damage probably by xanthine oxidase and oxygen species. MATERIALS AND METHODS: Isolated rat livers, lungs, and hearts were perfused with Krebs-Henseleit solutions. After stabilization, livers were either perfused or made ischemic. Then, livers and hearts or livers and lungs were reperfused in series, and the liver was disconnected and the second organ continued to perfuse with the accumulated effluents. MEASUREMENTS AND MAIN RESULTS: Ischemic and reperfused liver effluent contained high lactate dehydrogenase and uric acid concentrations compared with controls; xanthine oxidase increased 60 to 100 times. Ischemic and reperfused lung peak inspiratory pressure almost doubled; airway static compliance halved; myocardial contractility decreased to 70% of baseline; wet weight-to-dry weight ratios of lungs and livers increased. CONCLUSION: Ischemic and reperfused liver can directly induce myocardial and pulmonary dysfunction, presumably by oxidant induced injury. PMID- 9356060 TI - Leg tissue perfusion in simple tibial shaft fractures treated with unreamed and reamed nailing. AB - BACKGROUND: To compare the effects of unreamed and reamed intramedullary nailing on tibialis posterior, dorsalis pedis, and sum (tibialis posterior plus dorsalis pedis) distal arterial peak pulses. Additionally, leg skin temperature and transcutaneous oxygen tension were measured in patients with low energy, closed tibial shaft fractures. METHODS: The patients were randomized to unreamed and reamed groups, and intramedullary nailing without or with reaming was performed under spinal anesthesia. The measurements were carried out before the operation and on 5 postoperative days. RESULTS: In the unreamed group, the only significant difference between contralateral and nailed legs was in raised leg skin temperature (p = 0.0001). In the reamed group, tibialis posterior distal arterial peak pulses and transcutaneous oxygen tension remained at a significantly lower level and leg skin temperature at a significantly higher level, respectively, in the nailed legs after the operation when compared with contralateral legs (p = 0.0026, p = 0.0001, and p = 0.0001, respectively). There were no statistical differences between preoperative and postoperative values in the measured parameters in both groups. Additionally, there were no intergroup changes in the measured parameters in the injured legs. CONCLUSION: The present study suggests that altered distal arterial pulsations, decreased transcutaneous oxymetry values, and thermal reaction are not due to differences in nailing method but caused by a manifestation of the trauma mechanism of the tibial shaft fracture. The potentially negative effects of reaming to soft tissue perfusion parameters could not be established. PMID- 9356061 TI - Augmentative plate fixation for the management of femoral nonunion after intramedullary nailing. AB - Seventeen femoral nonunions after intramedullary nail internal fixation were treated with augmentative plate internal fixation. Six of them were initially managed with a Kuntscher nail internal fixation; the other 11 fractures were managed with a locked nail internal fixation. All the femoral nonunions were caused by insecure fixation of the intramedullary nailing, in which a rotational instability of the fracture site was verified in all cases during operation. Leaving the intramedullary nail in situ, an augmentative plate fixation was applied to the fracture site to counter the rotational instability. A simultaneous bone grafting was performed in seven of them to repair the bony defect. All these patients walked bearing full weight on the extremity without aching at the fracture site within 3 months and all these 17 fractures obtained a bony union within an average of 7 months after this treatment. From our experience, we have found this method is a useful treatment for the nonunion of the femoral shaft fracture after an intramedullary nail internal fixation. The technique is simple and does not require any special instrument. It facilitates an early weight bearing and gives a quick recovery from nonunion. PMID- 9356062 TI - Gunshot injuries to the brachial plexus. AB - OBJECTIVE: Gunshot wounds to the brachial plexus present a specific problem in peripheral nerve surgery. The purpose of this study is to analyze the characteristics of these injuries and the possibilities for functional recovery after their surgical treatment. DESIGN: Retrospective analysis of nerve lesions and results of surgery. MATERIALS AND METHODS: The series consists of 54 patients operated on, with 148 injured nerve elements, i.e., with 163 injured nerve elements when individual components of complex brachial plexus structures are included. Surgical procedures involved exploration, neurolysis, and nerve grafting, or their combination, depending on intraoperative findings. The surgical results were analyzed in 46 patients (85.1%) with 119 (80.1%) of the 148 nerve elements, with follow-up periods of more than 24 months. Standard grading scores for motor and sensory function were used and ranged according to functional priorities in the surgery of brachial plexus and individual nerves. MEASUREMENTS AND MAIN RESULTS: Neurolysis and nerve grafting generally gave similar rates of functional recovery, 90.2 and 87.8%, respectively. They were successful in all cases with injuries to the upper spinal nerves, the upper trunk, the lateral and posterior cord, and the musculocutaneous and axillary nerves. The failures were related to neurolysis or grafting of the ulnar and radial nerve lesions. CONCLUSION: Compared with previous studies, the number of lesions with complete functional loss and complete anatomic loss of continuity is larger. In cases that were prognostically favorable according to the location of injury, the results are similar regardless of the type of nerve repair. PMID- 9356064 TI - Cardiopulmonary effects of high-impulse noise exposure. AB - In high-energy impulse noise environments, the biomechanical coupling process between the external forces and the pathophysiology of cardiopulmonary injury is not well understood. A 12-in-diameter compressed air-driven shock tube with reflector plate was used to induce three levels of pulmonary contusion injury in a large animal model. Twenty-one anesthetized sheep were exposed to the various levels of impulse noise generated by the shock tube, with six additional sheep serving as a control group. Pathologic evaluations, performed 3 hours after exposure, showed pulmonary contusion ranging from minor petechial changes on the surface of the lung parenchyma to diffuse ecchymoses affecting as much as 60% of the lung. The gross pathologic observations of injury produced by exposure to the impulse noise produced by the shock tube were similar to those reported for blunt impact trauma or exposure to chemical or grain-dust explosions. The extent of lung injury (lung injury index) was quantitatively assessed. A semilogarithmic relationship between the lung injury index and the measured peak pressure was demonstrated. A significant linear correlation was demonstrated between lung injury index and lung weight-to-body weight ratio. Significant cardiopulmonary changes were also observed as a result of exposure to high-impulse noise. Although in most cases the degree of change was related to the severity of the injury, significant cardiopulmonary function changes were also observed in the absence of significant grossly observable pulmonary injury. Cardiac injury was indicated by decreased cardiac output and hypotension at all levels of injury and might be the result of myocardial contusion or air emboli. Pulmonary injury was demonstrated by respiratory acidosis, increases in lung resistance, and decreases in lung compliance and lung volume. Arterial PO2 appeared to be the most sensitive parameter of injury and was decreased for all measurement intervals for all exposure groups. PMID- 9356063 TI - Cardiopulmonary physiology of primary blast injury. AB - OBJECTIVE: Bomb blast survivors are occasionally found in profound shock and hypoxic without external signs of injury. We investigated the cardiovascular and pulmonary responses of rats subjected to a blast pressure wave. DESIGN: Prospectively randomized, controlled animal study. MATERIALS AND METHODS: Rats were instrumented and subjected to a blast pressure wave of different intensities from a blast wave generator. Cardiopulmonary parameters were recorded for 3 hours or until death. MEASUREMENTS AND MAIN RESULTS: The cardiovascular response to a blast pressure wave was immediate bradycardia, hypotension, and low cardiac index. Three hours later, the rats developed hypotension, low cardiac index, and low stroke volume. Interestingly, systemic vascular resistance remained unchanged. The pulmonary response was a decreased PaO2 and stable PacO2, suggesting a ventilation-perfusion mismatch from massive pulmonary hemorrhage. CONCLUSIONS: Blast-induced circulatory shock resulted from immediate myocardial depression without a compensatory vasoconstriction. Hypoxia presumably resulted from a ventilation-perfusion mismatch caused by pulmonary hemorrhage. PMID- 9356065 TI - Neurologic outcome with hemorrhagic hypotension after closed head trauma in rats: effect of early versus delayed conservative fluid therapy. AB - OBJECTIVE: This study examined (1) whether two previously reported, well established models in rats, one a model of hemorrhagic hypotension and the other a model of closed head trauma, could be combined to evaluate neurologic outcome when hemorrhage occurs subsequent to head injury, and (2) the ability of the traditional, conservative approach to fluid therapy (3 mL of intravenous fluid for 1 mL of blood loss) to reverse the detrimental effects of hemorrhagic hypotension after closed head trauma. In addition, two strategies of fluid therapy (early and delayed) were examined. METHODS: Fifty-six Sprague-Dawley male rats were divided into five groups with head injury at time 0 in groups 3 to 5, hemorrhage at 1 hour in groups 1, 2, 4, and 5, and intravenous fluid at 15 minutes (groups 2 and 5) or 60 minutes (groups 1 and 4) after hemorrhage. Head injury was delivered using a weight-drop impact of 0.5 J onto the closed cranium. Neurologic Severity Score (NSS) was determined at 1 hour (just before hemorrhage) and at 4 hours. RESULTS: NSS at 1 hour did not differ between groups 3 to 5 (15.5 (9-24) to 16 (2-21), median (range)). The amount of bleeding did not differ between groups during the first 15 minutes of hemorrhage (2.8 +/- 0.8 to 3.7 +/- 2.0 mL, mean +/- SD). After 60 minutes, cumulative blood loss in the delayed fluid therapy groups was less (3.1 +/- 1.13 mL in group 1 and 4.25 +/- 2.39 mL in group 4) than in the early fluid therapy groups (7.73 +/- 4.41 mL in group 2 and 6.85 +/- 2.36 mL in group 5) (analysis of variance, p < 0.01). The NSS of group 3 (head injury only) improved at 4 hours after injury (12 (5-20)), whereas the NSS of groups 4 and 5 (head injury followed by hemorrhage) deteriorated (24 (17-25) and 19.5 (9-25), respectively) (Kruskal-Wallis test,p < 0.05). In all the hemorrhage groups, fluid therapy failed to restore blood pressure to prehemorrhage levels. CONCLUSION: It is concluded that the two individual models of hemorrhagic hypotension and closed head trauma in rats can be combined to evaluate outcome when hemorrhage occurs subsequent to head injury. Furthermore, traditional, conservative fluid therapy, whether early or delayed, failed to restore blood pressure or to improve NSS when hemorrhage occurred after head injury. Blood loss was greater with early fluid therapy whether or not head injury was present. PMID- 9356066 TI - Effects of hemodilution on long-term survival in an uncontrolled hemorrhagic shock model in rats. AB - Prehospital guidelines for the treatment of penetrating trauma recommend rapid volume resuscitation to normal blood pressure. There is evidence, however, that fluid resuscitation to normal blood pressure in the setting of uncontrolled hemorrhagic shock (UHS) causes increased bleeding, hemodilution, and mortality. To test this hypothesis, we evaluated the effects of blood pressure and hemodilution on survival in a rat model of UHS. UHS was produced in rats by preliminary bleed of 3 mL/100 g followed by a 75% tail amputation. Experimental design consisted of three phases: a prehospital phase, with uncontrolled bleeding and resuscitation to either 40 or 80 mm Hg with lactated Ringer's solution (LR) or lactated Ringer's solution and whole blood (WB); followed by a hospital phase, with control of the bleeding and continued resuscitation to mean arterial pressure (MAP) > 80 mm Hg and hematocrit near 30%; followed by a 3-day observation phase. There were four treatment groups, n = 8 in each group: group I, MAP = 80 mm Hg with LR only; group II, MAP = 80 mm Hg with WB and LR; group III, MAP = 40 mm Hg with LR only; and group IV, MAP = 40 mm Hg with WB and LR. All group I rats died within 2.5 hours. There were no significant differences in survival among groups II, III, and IV. Base deficit, arterial pH, and lactate levels were significantly worse in the rats resuscitated to a MAP of 80 mm Hg with LR (group I). The effects of blood pressure alone, hemodilution alone, and their interaction were significantly related to base deficit and arterial pH. Hemodilution, but not blood pressure as an end point in resuscitation, was significantly related to lactate levels. The high mortality in this model of uncontrolled hemorrhage was attributable to the effects of blood pressure, hemodilution, and the interaction between the two variables, rather than simply continued blood loss from increased hydrostatic pressure. PMID- 9356068 TI - The use of hemorrhage occluder pins for controlling paravertebral intercostal artery bleeding: case report. AB - OBJECTIVE: To describe a technique for arresting traumatic bleeding uncontrollable by conventional means. METHODS AND RESULTS: Major intrathoracic bleeding was stopped in a 17-year-old boy who had been stabbed in his right chest by using a Hemorrhage Occluder Pin (Surgical Instruments Inc., Placetia, Calif). CONCLUSIONS: The use of occluder pins to stop bleeding from intercostal arteries may be life-saving. PMID- 9356067 TI - Effect of chronic cocaine exposure on the hemodynamic response to vasopressors in sheep. AB - BACKGROUND: The purpose of this study was to determine how chronic cocaine exposure affects the hemodynamic response to epinephrine, dopamine, phenylephrine, and ephedrine in awake sheep. METHODS: The hemodynamic response to dopamine (10 microg/kg), phenylephrine (1.5 microg/kg), and ephedrine (0.15 mg/kg) boluses was determined at baseline before low-dosage cocaine exposure and again after 15 and 18 days of cocaine exposure. The hemodynamic response to epinephrine (0.15 microg/kg), phenylephrine (1.5 microg/kg), and ephedrine (0.15 mg/kg) was determined at baseline before high-dosage cocaine exposure and again after 15 and 18 days of cocaine exposure. RESULTS: Chronic cocaine exposure abolished the mean arterial pressure and heart rate responses to dopamine but did not alter the responses to epinephrine, phenylephrine, or ephedrine. CONCLUSION: In awake sheep, chronic cocaine exposure markedly impairs the hemodynamic response to dopamine but not to epinephrine, phenylephrine, or ephedrine. PMID- 9356069 TI - Posterior fracture-dislocation of the shoulder with infraspinatus interposition: the buttonhole phenomenon. AB - We present a case with a posterior fracture-dislocation in which interposition of the infraspinatus precluded closed reduction by means of standard manipulation. The fractured greater tuberosity included the lesser tuberosity, allowing the infraspinatus to dislocate anterior to the dislocated humeral head and interpose between the humeral head and the glenoid cavity. The infraspinatus constricted the humeral neck together with the teres minor. Traction in the zero-position was thus required for resolution of constriction and subsequent reduction. Computed tomography was useful to demonstrate interposition of the infraspinatus and to plan the treatment. PMID- 9356070 TI - Proximal avulsion of the flexor carpi radialis muscle due to blunt trauma: a report of two cases. PMID- 9356071 TI - Blunt abdominal trauma causing chyloretroperitoneum. PMID- 9356072 TI - Acute tension diaphragmatic herniation: case report. PMID- 9356073 TI - Traumatic disruption of the innominate and right pulmonary arteries: case report. PMID- 9356074 TI - Profound hypotension in blunt trauma associated with superior gluteal artery rupture without pelvic fracture. AB - Superior gluteal artery injury is a rare but well-known complication of abdominal trauma, usually in association with pelvic fractures. Embolization has become the most effective treatment for pelvic hemorrhage with regard to superior gluteal artery injury, due to difficult surgical access. We report an unusual case of a superior gluteal artery rupture without pelvic fracture. The patient presented with profound hypotension after blunt trauma. Angiography revealed an injured superior gluteal artery, which was successfully embolized. PMID- 9356076 TI - Asymptomatic aortic stenosis and unexpected death in the trauma patient. AB - We report the cases of two young trauma patients with asymptomatic aortic stenosis who died after nonlethal blunt traumatic injuries. In both cases, their deaths were attributed to their underlying valvular disease. Awareness of the incidence of asymptomatic aortic stenosis and its potential physiologic hazard to the trauma victim may facilitate management of these difficult patients. PMID- 9356075 TI - Traumatic aortobiiliac dissection treated by kissing-stent placement. PMID- 9356078 TI - Where's the bullet? A migration in two acts. PMID- 9356079 TI - Wartime civilian injuries: epidemiology and intervention strategies. AB - BACKGROUND: Trauma is the most important public health risk in wartime. Most preventive effort have addressed the political etiology of armed conflicts and the secondary effects of war (food, water, shelter, sanitation, and vector control). Little to no efforts have addressed the direct prevention and control of war trauma. METHODS: An extensive review of the literature, with compilation of the most important data. RESULTS: Civilians are the major wartime targets in recent wars, and account for most of the killed and wounded. The trend has been toward a greater proportion of injuries from powerful explosive devices such as artillery shells and mines. Lessons learned from Bosnia and Lebanon show that the most effective way to achieve successful surveillance and injury prevention is to enhance the local skills and resources. CONCLUSIONS: New approaches are needed to minimize trauma to civilians. Both political advocacy and local efforts (including modifying firearms and ammunition, bullet proof helmets for children, anti-sniper shields) are needed. PMID- 9356077 TI - Aortoventricular fistula secondary to blunt trauma: a case report and review of the literature. AB - An aorto-right ventricular fistula secondary to nonpenetrating trauma is described. Review of the literature is reported. Ascending aortic injuries present as either traumatic pseudoaneurysms or, less commonly, as aortocardiac fistulas. Blunt cardiac injury is a frequent concomitant injury and contributes to the high mortality of this lesion. Prompt surgical intervention is required for survival. PMID- 9356080 TI - "Myocardial O2 balance during fluid resuscitation in uncontrolled hemorrhage: computer model". PMID- 9356081 TI - "Abdominal injuries without hemoperitoneum: a potential limitation of focused abdominal sonography for trauma (FAST)". PMID- 9356082 TI - "Conservative treatment of a traumatic tear of the left hepatic duct: case report". PMID- 9356083 TI - Acute normovolemic hemodilution can replace preoperative autologous blood donation as a standard of care for autologous blood procurement in radical prostatectomy. AB - Predonation of autologous blood (PAD) is a standard of care for patients undergoing radical prostatectomy, but recent studies have shown that PAD is not cost-effective. Acute normovolemic hemodilution (ANH) is an alternative autologous blood procurement technique that is much less costly than PAD. We compared the efficacy and costs of ANH alone to ANH combined with PAD. Two hundred-fifty patients who predonated fewer than 3 units of autologous blood before radical prostatectomy underwent ANH to a target hematocrit of 28%. Perioperative hematocrit levels, transfusion outcomes and costs, and postoperative outcomes were compared for patients who predonated 0, 1, or 2 units of blood before surgery. A computer model was used to estimate the savings in red blood cells (RBC) associated with each autologous intervention. ANH alone resulted in a 21% allogeneic transfusion rate and contributed a mean net savings of 112 mL RBC in blood conservation (equivalent to 0.6 unit of blood). The addition of 1 or 2 units of PAD reduced allogeneic exposure rates to 6% or 0%, respectively. Overall, patients who predonated blood had a mean net loss of 198 mL of RBC (equivalent to 1 blood unit), due to both an absence in compensatory erythropoiesis and to the wastage of 60% of the blood units donated. Patients who underwent ANH alone had a 60% reduction in mean total transfusion costs ($103 +/- $102) compared with patients who predeposited 2 units of autologous blood in addition to ANH ($269 +/- $11, P < 0.05). We conclude that ANH can replace PAD as an autologous blood option because it is less costly and equally effective. A combination of ANH and PAD can further decrease allogeneic blood exposure, but it increases transfusion costs and wastage. IMPLICATIONS: A patient's own blood can be obtained for use in surgery by predonation or acute normovolemic hemodilution on the day of surgery. Both blood collection techniques decrease the need for blood bank transfusions, but acute normovolemic hemodilution is less expensive and more convenient for patients. PMID- 9356084 TI - Fat elimination during intraoperative autotransfusion: an in vitro investigation. AB - Intraoperative autotransfusion of scavenged blood is an established method to reduce the need for perioperative homologous blood transfusion. However, if fat particles contaminate blood suctioned from the wound site, no reliable method is available to remove them during the washing and concentration of the recycled blood. A new generation of autotransfusion devices (e.g., continuous autotransfusion system [CATS]), based on separation chambers used in cell separators or plasmapheresis devices, allows continuous procession of the collected blood, in contrast with the discontinuous process used in conventional autotransfusion devices such as the Cell Saver 5. Theoretically, the continuous system should be more efficient than the discontinuous system in eliminating fat. Outdated, 36-day-old packed red blood cells, 600 mL, were mixed with 500 mL of lactated Ringer's solution and 200 mL of soya oil. Soya oil was used because it has a fatty acid composition similar to that of fat found in bone marrow. The blood mixture was then washed and concentrated by using either the CATS or the Cell Saver 5. Six samples were processed by each device. The CATS eliminated the soya oil (200 mL) completely, whereas the Cell Saver 5 delivered 30.3 +/- 7.8 mL soya oil into the retransfusion bag. The new generation of autotransfusion devices allows complete removal of fat particles. IMPLICATIONS: Autotransfusion devices serve to wash and retransfuse blood scavenged from the wound site. However, they cannot completely remove fat particles. This in vitro investigation showed that a new device completely removes fat particles and thus prevents retransfusion of fat. PMID- 9356085 TI - Tranexamic acid is effective in decreasing postoperative bleeding and transfusions in primary coronary artery bypass operations: a double-blind, randomized, placebo-controlled trial. AB - We evaluated the effects of tranexamic acid (TA) administered before and after cardiopulmonary bypass (CPB) in a prospective, randomized, placebo-controlled, double-blind study of adult patients undergoing primary coronary artery bypass grafting surgery. Patients received placebo (n = 30) or TA 15 mg/kg before CPB, followed by a TA infusion of 1 mg x kg(-1) x h(-1) for 5 h (n = 30) or TA 15 mg/kg after CPB, followed by a TA infusion of 1 mg x kg(-1) x h(-1) for 5 h (n = 30). Demographic, medical, surgical, laboratory, mediastinal chest tube drainage (MCTD), hemoglobin loss, transfusion, and outcome data were collected. Allogenic blood product administration was tightly controlled. The demographic, medical, and surgical characteristics were similar in all three groups. The median postoperative MCTD and hemoglobin loss in the pre-CPB TA group (710 mL, 8.6 g) was significantly less (P < 0.001) compared with the control (1202 mL, 44.2 g) and post-CPB TA groups (1020 mL, 23.4 g). The percentage of patients who received no allogenic blood products was 27% for the pre-CPB TA group and 33% for the post CPB TA group (not significant). These percentages were significantly lower than those in the placebo group (66%, P < 0.001). The median number of allogenic blood products administered to the pre-CPB TA group (0 units) was significantly less compared with the control group (4.5 units). The thromboelastogram and fibrinogen split product levels in the pre-CPB TA group indicated better platelet function and less activation of the fibrinolytic system compared with the other two groups (P < 0.05). There were no intergroup differences in reoperation, myocardial infarction, stroke, infections, or death. These data support the use of pre-CPB TA to decrease patient exposure to postcardiopulmonary bypass allogenic blood products. IMPLICATIONS: In this randomized, placebo-controlled trial, we investigated the efficacy of tranexamic acid to decrease bleeding and blood transfusions after open-heart operations. Tranexamic acid administered before and during the operation was effective in decreasing both bleeding and transfusions. When tranexamic acid was administered immediately after the operation, it had a minor beneficial effect. PMID- 9356087 TI - The effects of isoflurane and halothane on left ventricular afterload in dogs with dilated cardiomyopathy. AB - The effects of volatile anesthetics, including isoflurane (ISO) and halothane (HAL), on determinants of left ventricular (LV) afterload have not been comprehensively described in experimental models of, or patients with, heart failure. We tested the hypothesis that ISO and HAL produce beneficial alterations in LV afterload when evaluated with aortic input impedance and interpreted using a three-element Windkessel model in dogs before and after development of pacing induced cardiomyopathy. Hemodynamics and aortic pressure and blood flow waveforms were recorded in the conscious state and during 1.1- and 1.5-minimum alveolar anesthetic concentration (MAC) ISO and HAL anesthesia on separate days in chronically instrumented dogs (n = 6). Dogs were then paced at 220-240 bpm for 20 +/- 3 days (mean = SEM) to develop cardiomyopathy, and the experiments were repeated after pacing had been temporarily discontinued. ISO decreased mean arterial pressure (MAP), mean aortic blood flow (MAQ), and total arterial resistance (R) and increased total arterial compliance (C) and characteristic aortic impedance (Zc) in dogs before pacing. HAL decreased MAP and MAQ and increased C but did not alter R and Zc. Chronic rapid LV pacing increased HR and LV end-diastolic pressure and decreased MAP, LV systolic pressure, and the peak rate of increase of LV pressure. MAQ, C, R, and Zc were unchanged. ISO and HAL decreased arterial pressure but did not affect C and Zc in the presence of LV dysfunction. HAL, but not ISO, increased R at 1.1 MAC, which indicates that this drug increases resistance to LV ejection. In contrast to findings in normal dogs, these results indicate that neither ISO nor HAL reduce arterial hydraulic resistance to LV ejection or favorably improve the rectifying properties of the aorta in dogs with pacing-induced cardiomyopathy. IMPLICATIONS: Isoflurane and halothane produce favorable alterations in the determinants of left ventricular afterload before, but not after, the production of experimental left ventricular dysfunction by sustained, rapid cardiac pacing in chronically instrumented dogs. PMID- 9356086 TI - Light versus heavy sedation after cardiac surgery: myocardial ischemia and the stress response. Maritime Heart Centre and Dalhousie University. AB - The influence of light versus heavy sedation after coronary artery bypass graft (CABG) surgery on the development of postoperative myocardial ischemia has not been described. After uncomplicated CABG surgery, 50 patients were randomly assigned to receive LOW (n = 24; target Ramsay Sedation Score [RSS] = 2) or HIGH (n = 26; target RSS = 4) sedation with propofol. Analgesia was provided to maintain a visual analog scale (VAS) pain score <7. Myocardial ischemia was identified perioperatively using continuous 3-lead Holter monitoring. By measuring creatine kinase (CK) MB levels preoperatively, at entry to the intensive care unit (ICU), and every 12 h for 48 h; and by obtaining serial 12 lead electrocardiograms (ECG) (preoperatively; 2, 4, 12, 24, and 48 h after ICU admission, 8:00 AM the morning after surgery; and 5 min pre- and postextubation), myocardial infarction was identified. Endocrine stress response was assessed by measuring serum cortisol levels preoperatively, on admission to the ICU, and 24 h postoperatively. In a subset of patients (LOW n = 10, HIGH n = 11), plasma and urinary catecholamine levels were also measured. There were no between-group differences in demographics, operative course, hemodynamic variables, or cortisol levels while in the ICU. The VAS pain score and target RSS were achieved and sustained, and they differed between groups. There were three myocardial infarctions in each group by CKMB criteria alone. No ECG-identifiable myocardial infarction occurred. The ST segment versus time curve (LOW 187 +/- 295 versus HIGH 1071 +/- 2137 mm/min) differed between groups. Urinary and plasma catecholamine levels were similar between groups over the observation period. We conclude that the use of a reduced sedation regimen in combination with adequate analgesia did not result in an increased endocrine stress response or risk of myocardial ischemia. IMPLICATIONS: This randomized study of patients after coronary artery bypass surgery examined whether light (versus heavy) sedation with propofol in the intensive care unit was associated with an increased degree of myocardial ischemia. Using techniques to detect myocardial ischemia, including Holter monitoring, electrocardiogram, and myocardial enzyme measurements, no differences were found. We conclude that light sedation does not increase the endocrine stress response or the risk of myocardial infarction. PMID- 9356088 TI - Augmenting cardiac contractility hastens myocardial edema resolution after cardiopulmonary bypass and cardioplegic arrest. AB - Although myocardial edema is associated with cardiopulmonary bypass (CPB) and cardioplegic arrest (CPA), interventions to expedite edema removal have not been investigated. The primary mechanism for the removal of excess interstitial fluid in the heart is myocardial lymphatic drainage, but lymphatic function can be impaired by decreased contractility because of edema. The purpose of this study was to determine whether enhancing cardiac contractility would increase myocardial lymphatic function and hasten edema resolution after CPB. Sixteen dogs were subjected to CPB and 1 h of hypothermic CPA. After weaning from CPB, 10 dogs received an intravenous dobutamine infusion and 6 dogs received no inotropic support. We determined myocardial lymph driving pressure from the major cardiac lymphatic, myocardial water content by using microgravimetry, and the peak rate of left ventricular pressure increase (dP/dmax) by using micromanometry. Measurements were taken at baseline, during CPA, and 60 min after CPB. Compared with controls, dobutamine-treated dogs had an increased dP/dmax (P < 0.05), which was associated with higher lymph driving pressures (P < 0.05), resulting in lower myocardial water gain 1 h after CPB (P < 0.05). We conclude that the resolution of myocardial edema after CPB was hastened by dobutamine. Organized ventricular contraction and myocardial contractility seem to be important determinants of myocardial lymphatic function and myocardial edema removal. These findings suggest that the administration of inotropic drugs after CPB may hasten cardiac recovery. IMPLICATIONS: Myocardial edema, which develops during cardiopulmonary bypass and cardioplegic arrest, contributes to cardiac dysfunction after heart surgery. This study demonstrated that enhancement of cardiac contractility by the administration of dobutamine after cardiopulmonary bypass and cardioplegic arrest was associated with increased myocardial lymphatic function and hastened edema resolution in dogs. PMID- 9356089 TI - Myocardial ischemia and adverse cardiac outcomes in cardiac patients undergoing noncardiac surgery with sevoflurane and isoflurane. Sevoflurane Ischemia Study Group. AB - Sevoflurane is associated with less tachycardia and coronary vasodilation than isoflurane and thus might be associated with less myocardial ischemia. This multicenter study examined the incidence of myocardial ischemia and adverse cardiac outcomes in adults (40-87 yr) with cardiac disease having elective noncardiac surgery. Patients were randomized to receive either sevoflurane (S) (n = 106) or isoflurane (I) (n = 108) in conjunction with sodium thiopental, vecuronium, fentanyl, and 50%-70% N2O. Intraoperative hemodynamics were maintained within 20% of awake baseline with standard drugs. A Holter monitor was applied 3-24 h before surgery and maintained until 48 h after surgery. Electrocardiograms and blood samples for analysis of the MB isoenzyme fraction of creatine phosphokinase were obtained preoperatively and daily for 48 h postoperatively. Anesthetic exposure (1.79 +/- 0.15 [mean +/- SE] minimum alveolar concentration-hour) and duration of surgery (219 +/- 13 min) did not differ between groups. The incidence of ischemia in the pre-, intra- and postoperative periods, adverse cardiac outcomes (18% occurrence), intraoperative hemodynamic variations (+/-20% change from ward baseline), and administration of adjunct cardiovascular medications were similar between groups. In cardiac patients having noncardiac surgery, sevoflurane was comparable to isoflurane with respect to the incidence of intra- and postoperative myocardial ischemia and in the frequency of adverse cardiac outcomes. IMPLICATIONS: Surgical patients with heart disease are at risk of heart complications, some of which could be induced by an anesthetic. We compared the incidence of cardiac complications between patients receiving sevoflurane and isoflurane. We found that the frequency of additional heart problems in cardiac patients receiving sevoflurane was not different from that associated with isoflurane. PMID- 9356090 TI - The vasodilator effects of clevidipine on human internal mammary artery. AB - Endothelial dysfunction and platelet activation with thromboxane release may contribute to spasm or alterations in internal mammary artery (IMA) graft flow during coronary artery surgery. Clevidipine, an ultrashort-acting dihydropyridine calcium channel blocker, is undergoing clinical development, but there are little data regarding its effects on human vasculature. We investigated the effects of clevidipine on human IMA obtained during surgery. After precontracting IMA segments with an analog of thromboxane (U46619, 10(-8) mol/L), acetylcholine and nitroglycerin were added cumulatively to examine endothelial function. Concentration-response curves to clevidipine were cumulatively obtained during submaximal contraction to the U46619 (10(-8) mol/L) in rings with and without endothelium. In the IMA samples with endothelium, acetylcholine did not completely reverse the U46619-mediated contraction, which implies impaired endothelial function (40% +/- 6% maximal response). Both clevidipine and nitroglycerin completely reversed U46619-induced contraction (clevidipine (50% effective concentration [EC50] = 3.88 +/- 0.84 x 10(-6) mol/L, nitroglycerin EC50 = 4.84 +/- 2.76 x 10(-8) mol/L). The responses to clevidipine were similar in preparations with or without intact endothelium. Clevidipine is an endothelium independent arterial vasodilator that offers a potential therapeutic option in the treatment of perioperative arterial graft vasospasm and/or hypertension. IMPLICATIONS: Clevidipine is a new ultrashort-acting dihydropyridine calcium antagonist. In human internal mammary arteries precontracted with a thromboxane A2 analog, clevidipine was an effective vasodilator on vessel segments in the presence and in the absence of endothelium. PMID- 9356092 TI - Long Q-T syndrome associated with oral erythromycin used in preoperative bowel preparation. PMID- 9356091 TI - Attenuation of cardiovascular responses to tracheal extubation: comparison of verapamil, lidocaine, and verapamil-lidocaine combination. AB - We recently showed that verapamil attenuated hemodynamic responses to tracheal extubation. The aim of the current study was to compare the efficacy of a combination of intravenous (I.V.) verapamil (0.1 mg/kg) and I.V. lidocaine (1 mg/kg) with that of each drug alone in suppressing the cardiovascular changes during tracheal extubation and emergence from anesthesia. One hundred adult patients (ASA physical status I) who were to undergo elective minor surgery were randomly assigned to one of four groups (n = 25 each): Group S = saline plus saline (control), Group V = verapamil 0.1 mg/kg I.V. plus saline, Group L = lidocaine 1 mg/kg I.V. plus saline, and Group V-L = verapamil 0.1 mg/kg I.V. plus lidocaine 1 mg/kg I.V. These medications were given 2 min before tracheal extubation. Anesthesia was maintained with 1.0%-2.0% sevoflurane and 60% nitrous oxide (N2O) in oxygen. Muscle relaxation was achieved with vecuronium, and a residual neuromuscular blockade was reversed with neostigmine 0.05 mg/kg (combined with atropine 0.02 mg/kg). Changes in heart rate (HR) and arterial blood pressure (AP) were measured during and after tracheal extubation. In the control group, the HR and systolic and diastolic AP increased significantly during tracheal extubation. Verapamil, lidocaine, and their combination attenuated the increases in these variables. The beneficial effect was the greatest with the combination of verapamil and lidocaine. These findings suggest that verapamil 0.1 mg/kg and lidocaine 1 mg/kg given I.V. concomitantly 2 min before tracheal extubation is a simple and more effective prophylaxis than verapamil or lidocaine alone for attenuating the cardiovascular changes associated with tracheal extubation. IMPLICATIONS: Tachycardia and hypertension associated with tracheal extubation, which may lead to myocardial ischemia, represent a potential risk for patients with coronary arterial disease. To seek effective pharmacological prophylaxis against these complications, we compared the attenuation of hemodynamic changes among verapamil, lidocaine, and a verapamil/lidocaine combination using ASA physical status I patients and found the combination to be effective. PMID- 9356093 TI - A randomized, blind comparison of remifentanil and alfentanil during anesthesia for outpatient surgery. AB - We compared remifentanil, an esterase-metabolized opioid, with alfentanil as part of balanced anesthesia with at least 0.8% isoflurane during outpatient surgery in a randomized, double-blind trial. One hundred two patients received remifentanil, and 99 patients received alfentanil. Patients who received remifentanil experienced significantly fewer stress responses to surgical stimuli (52.9% and 65.7%, P < 0.05); significantly fewer remifentanil patients responded to skin closure (11% and 22%, P < 0.05) than patients who received alfentanil. Significantly more patients in the alfentanil group required extra analgesia compared with the remifentanil group (P < 0.05). Time to respond to verbal command was shorter for alfentanil than remifentanil (median 7 min vs 9 min), and times to spontaneous respiration (median 5 min vs 8 min), adequate respiratory rate (median 6 min vs 9 min), and tracheal extubation (median 6 min vs 9 min) were significantly shorter for alfentanil in comparison with remifentanil (P < 0.05). Remifentanil patients, however, showed significantly better recovery of psychomotor and psychometric function between 30 and 90 min after surgery (P < 0.05). The incidences of hypotension intraoperatively and shivering postoperatively were significantly higher with remifentanil. No unexpected or serious adverse events were recorded with remifentanil; however, one patient who received alfentanil experienced severe recurrent respiratory depression after surgery. The metabolic profile of remifentanil allowed better intraoperative analgesia without compromising recovery. IMPLICATIONS: The pharmacological profile of remifentanil, a new opioid for use in anesthesia, suggests that rapid recovery will occur after its use. This study of 200 outpatients shows that the differences suggested from kinetic studies are not always borne out in clinical practice, although later recovery variables did, in fact, favor remifentanil. PMID- 9356094 TI - The changing role of monitored anesthesia care in the ambulatory setting. PMID- 9356095 TI - Teaching the use of fiberoptic intubation for children older than two years of age. AB - In 144 anesthetized children aged 2-9 yrs, the safety and feasibility of orotracheal fiberoptic intubation, with and without an airway endoscopy mask, were assessed and compared with laryngoscopic intubation. Eight anesthesia residents with experience in adult fiberoptic intubation, but who were beginners in pediatric anesthesia, participated in this study. In a randomized fashion, each resident intubated 18 children (6 in each group). The time (mean +/- SD) to achieve successful intubation was different for laryngoscopic and fiberoptic intubation (34 +/- 17 s and 80 +/- 39 s, respectively; P < 0.001). The use of the airway endoscopy mask further prolonged fiberoptic intubation (167 +/- 121 s, P < 0.001). Spo2 values remained >95% in all patients during conventional laryngoscopy and fiberoptic laryngoscopy with a mask, whereas Spo2 decreased below 95% in 2 of the 48 patients during fiberoptic intubation without a mask. Both patients promptly recovered during ventilation via a face mask. We conclude that teaching the use of fiberoptic intubation in healthy, anesthetized children aged 2-9 yrs is safe and feasible. IMPLICATIONS: Fiberoptic intubation is a valuable technique of airway management. We studied the feasibility and safety of a training program that could be used for children more than 2 yrs old. This study demonstrates that fiberoptic intubation can be effectively practiced in pediatric patients without increased risk of side effects. PMID- 9356096 TI - Anesthesia for pheochromocytoma in a surgically anephric child. PMID- 9356097 TI - A new biological assay for measuring cyanide in blood. AB - Clinical diagnosis of cyanide poisoning is complicated by the lack of an easy, convenient assay for cyanide concentration in blood. Therapy may be delayed with unconfirmed diagnosis because the conventional antidote to cyanide poisoning exposes patients to substantial risks. We developed a new spectrophotometric assay to measure cyanide by extraction into a sodium hydroxide trap, followed by the addition of exogenous methemoglobin as a colormetric indicator. Samples of blood from 15 healthy subjects and 5 patients who had received prolonged nitroprusside infusions were assayed. To optimize assay characteristics, methemoglobin concentrations, pH, temperature, incubation time, and buffer strengths were varied. Duplicate samples were assayed by using the polarographic method for assay validation. Over a range from 300 ng/mL to 7 microg/mL, the correlation between methods was r = 0.983. Interassay and intraassay variability were 5% and 2%, respectively. Samples drawn from the five patients and tested by using both methods yielded a correlation of r = 0.978. This new assay for cyanide in blood may greatly facilitate the diagnosis and treatment of cyanide ingestion. The use of methemoglobin as the colorimetric indicator in the assay contributes to its low cost and ease of use. IMPLICATIONS: Cyanide, an important factor in death from burn-related inhalation injury, is difficult and time-consuming to measure. We developed a new, rapid blood test for cyanide using methemoglobin as a colormetric indicator. A rapid, accessible test for cyanide may speed the diagnosis and treatment of cyanide poisoning. PMID- 9356098 TI - Acute pansialadenopathy during induction of anesthesia causing airway obstruction. PMID- 9356099 TI - The differential effects of prostaglandin E1 and nitroglycerin on regional cerebral oxygenation in anesthetized patients. AB - We evaluated the effects of prostaglandin E1 (PGE1) and nitroglycerin (NTG) on regional tissue oxygenation and use in the brain using near infrared spectroscopy (NIRS). Twenty-four patients who underwent elective cardiac surgery were randomly divided into two groups. The study was performed after the induction of anesthesia and before the start of the surgical procedure. After measuring arterial and jugular venous blood gases, cardiovascular hemodynamics, and relative cerebral oxyhemoglobin (HbO2), deoxyhemoglobin, and cytochrome aa3 at the baseline, PGE1 (n = 12) or NTG (n = 12) was infused intravenously at a rate of 0.3 g/kg or 5 g/kg, respectively. Thirty minutes after the start of drug infusion, administration of the drugs was stopped. Both PGE1 and NTG reduced mean arterial pressure to approximately 70% of the baseline value 10, 20, and 30 min after start of drug infusion (P < 0.05). Internal jugular venous pressure increased significantly during NTG but not during PGE1 infusion (P < 0.05). PGE1 increased HbO2 concentration, which was sustained for 30 min after discontinuing the drug. NTG increased HbO2 concentration, but this gradually returned to the baseline level after discontinuation of the drug. Baseline value of jugular oxygen saturation was 64.5% +/- 2.1%, and there was no significant changes during the infusion of PGE1 or NTG. These results demonstrate that both NTG and PGE1 increased cerebral oxygen saturation as measured by NIRS. This may be explained by local cerebral hyperemia without major alteration in flow/metabolism coupling of brain. The onset of this increase was slower and the duration of this effect after discontinuation of the drug was more prolonged with PGE1. These phenomena occurred despite the relatively similar time course of the effect of these two drugs on systemic hemodynamic values. IMPLICATIONS: The cerebrovascular effects of vasodilators used for induced hypotension are not fully understood. In this study, we used near infrared spectrometry and jugular oxygen saturation measurement to assess the effects of prostaglandin E1 and nitroglycerin on cerebral perfusion. We found that nitroglycerin and prostaglandin E1 increase cerebral oxygen saturation as measured by near infrared spectrometry, but with different time courses. This information will hopefully help anesthesiologists to better maintain adequate regional cerebral oxygenation. PMID- 9356100 TI - Risk factors for neurologic deterioration after revascularization surgery in patients with moyamoya disease. AB - To investigate the risk factors for postoperative neurological deterioration in patients with moyamoya disease, we retrospectively reviewed the perioperative course of 368 cases of revascularization surgery in 216 patients with this disease. Risk factors anecdotally associated with postoperative ischemic events were analyzed by comparing groups with or without a history of such events on the operative day. Ischemic events were noted in 14 cases (3.8%), 4 of which were defined as strokes and the others as transient ischemic attack (TIA). Postoperative neurological deterioration more often developed in patients who suffered from frequent TIAs, had precipitating factors for TIA, and underwent indirect nonanastomotic revascularization. The authors conclude that the incidence of postoperative ischemic events were related more to the severity of moyamoya disease and the type of surgical procedure than to other factors, including anesthetic management. IMPLICATIONS: Although preventing stroke is the major concern for patients with moyamoya disease, risk factors for perioperative cerebral ischemia have not been clarified. We retrospectively analyzed the perioperative course in 368 cases with this disease and found that the severity of the disease and type of surgical procedure were major determinants of postoperative cerebral ischemia. PMID- 9356101 TI - Is epidural anesthesia in labor associated with chronic low back pain? A prospective cohort study. AB - The association between epidural anesthesia during labor and subsequent postpartum low back pain remains unclear. The objective of this follow-up cohort study was to determine whether epidural anesthesia was associated with chronic back pain 1 yr after delivery. We contacted 329 women by telephone and asked them to complete a standardized questionnaire 1 yr (+/-1 mo) after delivery. One hundred sixty-four women had received epidural analgesia for labor and delivery, and 165 had not. Subjects were asked to quantify their back pain (yes/no, numeric rating score, and interference with daily activities). Differences between the two groups were tested by using the chi2 test and the Mann-Whitney U-test, and logistic regression was used to control for confounding variables. The response rate was 244 of 329 (74%). Responders and nonresponders were similar in their demographic and clinical characteristics. There was no difference in the prevalence of back pain between women who had received epidural anesthesia (12 of 121, 10%) and those who had not (17 of 123, 14%). The adjusted relative risk of low back pain at 1 yr (epidural versus nonepidural) was 0.63 (95% confidence interval 0.25, 1.56). There were also no differences between the two groups on numeric rating scores or level of interference with activities. This prospective follow-up study demonstrated no association between epidural anesthesia for labor and delivery and chronic back pain 1 yr after delivery. IMPLICATIONS: We evaluated the presence of low back pain 1 yr after delivery in two groups of women-those who chose epidural analgesia for labor and those who did not. There was no increased risk of back pain in women who had used epidural analgesia. This finding is consistent with those of other North American studies. PMID- 9356102 TI - Anesthetic management of a parturient with olivopontocerebellar degeneration. PMID- 9356103 TI - Management of nonobstetric pain during pregnancy and lactation. PMID- 9356104 TI - Risk of respiratory arrest after intrathecal sufentanil. PMID- 9356105 TI - The effect of age on retrieval of local anesthetic solution from the epidural space. AB - We conducted this prospective study to determine whether advancing age is correlated with retrieval of local anesthetic solution from the epidural space. Three hundred forty-six patients (ASA physical status I or II, 20-93 yrs of age, 177 female and 169 male patients) undergoing epidural anesthesia were enrolled. The epidural space was identified by a loss of resistance technique using air, and a catheter was introduced 3 cm. Three milliliters of 2% lidocaine with epinephrine was injected as a study dose by hand at a rate of 1 mL/s with the patient in the supine position. The syringe was immediately aspirated to retrieve the local anesthetic solution. A retrieved volume of 0.5 mL or more with a glucose concentration less than 6 mg/dL was defined as retrieval positive, and a volume of less than 0.5 mL was defined as retrieval negative. There was a significant correlation between age and retrieval volume among all the patients (Y = 0.008X-0.222, P < 0.0001) with a significant increase in the positive retrieval incidence and volume from the patients in their 50s (11%, 0.6 +/- 0.3 mL) to the patients in their 60s (26%, 1.0 +/- 0.6 mL) (P < 0.05 for both). The incidence of positive retrieval and the retrieval volume were greater in the patients in their 60s and older (30%, 1.1 +/- 0.63 mL) than in the younger than 60 (10%, 0.6 +/- 0.3 mL) (P < 0.0001 and P < 0.001). The glucose concentration was 2.3 +/- 1.2 mg/dL in the positive cases. We conclude that there is a weak positive correlation between age and the local anesthetic solution retrieved from the epidural space. IMPLICATIONS: We conducted a study in 346 patients to determine whether advancing age could be correlated with retrieval of local anesthetic solution from the epidural space. We found a weak positive correlation between advanced age and the amount of solution retrievable from the epidural space. Further studies are required to determine whether this phenomenon may call for dose adjustments in patients aged more than 60 yrs. PMID- 9356106 TI - The effect of epidural saline injection on analgesic level during combined spinal and epidural anesthesia assessed clinically and myelographically. AB - An epidural injection of physiological saline solution after spinal anesthesia may produce a higher level of analgesia than spinal anesthesia alone because of a volume effect. The purpose of this study was to clarify the volume effect caused by epidural injection of saline after spinal anesthesia. Twenty patients undergoing combined spinal and epidural anesthesia for elective surgery whose analgesic levels did not reach the surgical regions 10 min after spinal anesthesia at the L4-5 interspace were randomly assigned to two groups. The control group (n = 10) received no epidural saline injection. The saline group (n = 10) received 10 mL of saline through an epidural catheter at the L2-3 or L3-4 interspace 10 min after spinal anesthesia. In the saline group, the levels of analgesia 15 and 20 min after spinal anesthesia were significantly higher than those in the control group (P < 0.05). Next, we examined the volume effect of epidural injection of saline with myelography using two adult volunteers. In both volunteers, the upper level of the contrast medium, which was injected in the lumbar subarachnoid space, began to increase concurrently with lumbar epidural injection of saline, reaching from L3 to L1 and from L2 to T12. The diameter of the subarachnoid space diminished to less than 25% after injection of saline. We conclude that lumbar epidural injection of saline increases the analgesic level 10 min after spinal anesthesia, probably because of a volume effect. IMPLICATIONS: In this study, using surgical patients and volunteers, we determined that a lumbar epidural injection of physiological saline solution 10 min after spinal anesthesia produces a higher analgesic level than spinal anesthesia alone because of a volume effect. PMID- 9356107 TI - A microscopic analysis of cut-bevel versus pencil-point spinal needles. AB - An in vitro examination of 25-gauge Quincke and 25-gauge and 27-gauge Whitacre spinal needles was performed after insertion in 210 consenting adult patients. In addition, 300 unused Quincke needles and 300 unused pencil-point needles were examined under a dissecting microscope. When the microscopic evaluation was performed on the needles after spinal blockade, burrs or blunting of the needle tip were noted in 24% of the Quincke needles compared with only 3% of the 25 gauge Whitacre needles and 10% of the 27-gauge Whitacre (P < 0.05). Bony contact with 25-gauge Quincke and 27-gauge Whitacre needles resulted in an increased incidence of microscopic tip damage (versus 25-gauge Whitacre). Needle-tip damage with the Whitacre needles was limited to blunting of the tip. The analysis of unused needles revealed significant differences among manufacturers of the cut bevel needles with respect to stylet-to-needle length and burrs on the end of the stylet. The leading edge of the stylet protruded beyond the opening of the needle tip in 7% of the Quincke needles. However, only minor needle-tip abnormalities were noted with the pencil-point needles (i.e., variability in the side-port opening to needle tip distance, side-port opening integrity). In conclusion, bony contact produced more damage to the cut-bevel than to the pencil-point needle tips. In addition, fewer inherent manufacturing defects were noted with the pencil-point versus cut-bevel needles. IMPLICATIONS: It has been suggested that damaged needle tips may contribute to a higher incidence of headaches after spinal anesthesia. A microscopic examination revealed that the pencil-point (versus cut-bevel) needles had fewer manufacturing flaws and were less susceptible to tip damage when bony contact occurred during the placement of the spinal needle. PMID- 9356108 TI - A comparative efficacy study of hyperbaric 5% lidocaine and 1.5% lidocaine for spinal anesthesia. AB - We compared the clinical efficacy of 1.5% lidocaine in dextrose 7.5% in water, which is currently available as a commercial preparation but approved for use only in obstetrical patients, with the traditional 5% lidocaine in dextrose 7.5% in water for spinal anesthesia in patients undergoing lower abdominal procedures. Fifty-one male patients scheduled to undergo inguinal herniorrhaphy were randomly divided into two groups based on the spinal anesthetic received: Group I received 1.5% lidocaine in dextrose 7.5% in water, and Group II received 5% lidocaine in dextrose 7.5% in water. After intrathecal injection of the anesthetic, each patient was evaluated for the speed of onset, the time to recovery, and the quality of the surgical anesthesia and motor block that ensued by an anesthesiologist blinded to the technique. With the exception of the patients in Group I, who achieved a higher dermatome level of sensory analgesia, we were unable to demonstrate any significant clinical differences between the two lidocaine solutions. Our results indicate that lidocaine 1.5% in dextrose 7.5% in water is clinically indistinguishable from the 5% solution as a spinal anesthetic for lower abdominal surgery. IMPLICATIONS: In this study, two concentrations of lidocaine are compared as spinal anesthetics in 51 male patients undergoing inguinal hernia repair. Patients were assessed for the onset, quality, and duration of the spinal block. The study results indicate that 1.5% lidocaine is as effective as the 5% solution as a spinal anesthetic for patients undergoing inguinal hernia repair. PMID- 9356109 TI - Nitrous oxide enhances the level of sensory block produced by intrathecal lidocaine. AB - We examined the effect of nitrous oxide (N2O) administration on the level of sensory block produced by intrathecal lidocaine in patients undergoing transurethral procedures. Twenty minutes after subarachnoid injection of 100 mg (5%) hyperbaric lidocaine, the level of block to pressure sensation was assessed. After establishing the baseline sensory block, patients were randomly assigned to receive either 50% nitrogen (control group) or 50% N2O in oxygen for 10 min, and the sensory level was reassessed. All patients then received 35% oxygen for 5 min, and the level of block to pressure was assessed again. Changes were measured in centimeters and standardized by dividing those results by the height of patients (in centimeters). Ten minutes after nitrogen or N2O administration, a 3.8-cm regression of sensory block was found in the control group, and a 1.8-cm cephalad increase was found in the treatment group (P < 0.0001). Discontinuation of N2O for 5 min resulted in a rapid regression of the level of sensory block (4 cm in the N2O group versus 1.9 cm in the control group, P < 0.0001). However, 5 min after discontinuation of N2O, the overall regression of the sensory block in the control group, when measured from the baseline, was 5.7 cm versus 2.2 cm in the N2O group (P < 0.001). The differences between the two groups before standardization are consistent with those after standardization (t = 9.02 at 10 min, t = 4.24 at 15 min, and t = 3.97 for the overall change at 15 min). The results suggest that inhalation of 50% N2O enhances the level of sensory block produced by intrathecal lidocaine. IMPLICATIONS: We measured the level of sensory block produced by subarachnoid anesthesia with lidocaine before and after inhalation of 50% nitrous oxide for 10 min. Nitrous oxide enhanced the level of subarachnoid anesthesia with minimal hemodynamic effects. These findings are of clinical importance when subarachnoid anesthesia subsides before the completion of surgery. PMID- 9356110 TI - The effect of somatostatin on experimental inflammation in rats. AB - The aim of the present study was to investigate the effect of somatostatin administration on experimentally induced inflammation in rats. Inflammation was induced by the intraplantar injection of carrageenan (50 microL) into the hind paw of the rat. Animals were treated intraplantarly with somatostatin in a volume of 50 microL at different doses (2.5, 25, and 250 ng, 10 microg). The inflammatory response was studied 120, 180, and 240 min after drug administration. The antinociceptive effect of somatostatin was determined by using the Randall and Selitto test and by local production of beta-endorphin from lymphocytes obtained from popliteal lymph nodes. Data show that small doses of somatostatin were the most effective in reducing hyperalgesia. Moreover, our results show that somatostatin treatment significantly increased beta-endorphin in lymphocytes from popliteal lymph nodes. The secretion of opioid peptides, which enhance analgesia, could be stimulated by locally administered somatostatin. IMPLICATIONS: Acute pain because of intraplantar inflammation induced in rats by carrageenan injection was significantly reduced by small-dose, local administration of somatostatin, which possibly favors beta-endorphin release as a mechanism. These results may have implications regarding treatment of pain conditions associated with an inflammatory response. PMID- 9356111 TI - Ondansetron exhibits the properties of a local anesthetic. AB - The purpose of this study was to determine whether ondansetron (OND) has local anesthetic effects. Using a patch-clamp technique, we showed that OND concentration dependently blocked Na channel currents in freshly isolated neurons of rat brains with a 50% inhibition concentration of 12 microM. The blockade started immediately when OND was applied to the cell body using a fast perfusion system, reached a plateau within 15 s, and recovered to the control level within 30 s after washout of the OND-containing solution. Because this is a known property of local anesthetics, we used the tail-flick technique to verify this effect in vivo in Sprague-Dawley rats (n = 46). OND was injected subcutaneously into the tail at the doses of 0.08, 0.16, and 0.2 mg. The tail-flick latency increased 2 min after OND injection, reaching the plateau within 5 min. This effect was dose-related, lasting from 10 to 25 min. These preliminary data indicate that OND, a selective 5-HT3 receptor antagonist, might serve as a prototype molecule for development of a novel series of local anesthetics. IMPLICATIONS: Ondansetron is a drug used to prevent vomiting, especially in cancer patients after chemotherapy. We found that it also causes numbness when injected under the skin. This new action may contribute to its role in "calming the stomach." We studied the effect of ondansetron on the isolated brain cells of live rats. PMID- 9356112 TI - Peripherally administered alpha2-adrenoceptor agonist in the modulation of chronic allodynia induced by spinal nerve ligation in the rat. AB - We studied whether a peripherally administered alpha2-adrenoceptor agonist modulates mechanical allodynia caused by unilateral ligation of two spinal nerves in the rat. Medetomidine, an alpha2-adrenoceptor agonist, atipamezole, an alpha2 adrenoceptor antagonist, or saline (control) was administered into the footpad of either the allodynic or the contralateral hindpaw. Medetomidine (1-10 microg/kg in 50 microL) reversed the unilateral allodynia in a dose-dependent fashion independent of the site of administration. At this dose range, medetomidine did not influence the heat-induced tail-flick response. The antiallodynic effect of medetomidine was completely reversed by a dose of atipamezole that alone was ineffective (30 microg/kg). At the largest dose used (100 microg/kg in 50 microL), atipamezole decreased the latency of the heat-induced tail flick and had an ipsilateral allodynic effect in the injected paw when administered into the control side. The atipamezole-induced mechanical allodynia was not attenuated by medetomidine. The results indicate that an alpha2-adrenoceptor agonist at a subantinociceptive dose may significantly attenuate allodynia produced by spinal nerve ligation. This antiallodynic effect is not due to a peripheral action, but rather to action on central (presumably spinal) alpha2-adrenoceptors. The allodynic effect of a high concentration of atipamezole (100 microg/kg in 50 microL) in the injected paw can be explained by peripheral nonadrenergic mechanisms (e.g., local irritation). IMPLICATIONS: The present behavioral results indicate that a selective alpha2-adrenoceptor agonist at a subantinociceptive dose effectively attenuates mechanical allodynia induced by an experimental model of chronic neuropathy in the rat. This antiallodynic action can be explained by central, rather than peripheral, alpha2-adrenergic mechanisms. PMID- 9356113 TI - Convulsions induced by ropivacaine during interscalene brachial plexus block. PMID- 9356114 TI - The effects of inhaled nitric oxide and its combination with intravenous almitrine on Pao2 during one-lung ventilation in patients undergoing thoracoscopic procedures. AB - The aim of this study was to assess whether hypoxemia during one-lung ventilation (OLV) can be prevented by inhaled nitric oxide (NO) (Part I) or by its combination with intravenous (IV) almitrine (Part II) in 40 patients undergoing thoracoscopic procedures. In Part I, 20 patients were divided into two groups: one received O2 (Group 1) and one received O2/NO (Group 2). In Part II, 20 patients were divided into two groups: one received O2 (Group 3) and one received O2/NO/almitrine (Group 4). In Groups 2 and 4, NO (20 ppm) was administered during the entire period of OLV, and almitrine was continuously infused (16 microg x kg( 1) x min[-1]) in Group 4. Arterial blood gases were measured during two-lung ventilation with patients in the supine position, after positioning in the lateral decubitus position, and then every 5 min for a 30-min period during OLV. During OLV, Pao2 values decreased similarly in Groups 1 and 2. After 30 min of OLV, the mean Pao2 values in Groups 1 and 2 were 132 +/- 14 mm Hg (mean +/- sem) and 149 +/- 27 mm Hg (not significant [NS]), and the Pao2 value was less than 100 mm Hg in four patients in Group 1 and five patients in Group 2. Pao2 values were greater in Group 4 than in Group 3 after 15 and 30 min of OLV. After 30 min of OLV, the mean Pao2 values were 146 +/- 16 mm Hg in Group 3 and 408 +/- 33 mm Hg in Group 4 (P < 0.001). Pao2 was less than 100 mm Hg during OLV (NS) in four patients in Group 3 and in no patient in Group 4. We conclude that NO inhalation alone has no effect on Pao2 evolution during OLV, although its combination with IV almitrine limits the decrease of Pao2 during OLV. This beneficial effect of NO/almitrine could be attributed to an improvement in ventilation-perfusion relationships. IMPLICATIONS: Decrease in oxygenation during one-lung ventilation is quite common. Our study showed that inhaled nitric oxide alone did not influence Pao2 evolution. We then tried adding intravenous almitrine to nitric oxide with amazingly good results on Pao2. This nonventilatory technique should be of great use during special thoracic acts, such as thoracoscopic procedures. PMID- 9356115 TI - Postoperative pharmacokinetics and sympatholytic effects of dexmedetomidine. AB - Dexmedetomidine is a selective alpha2-adrenoceptor agonist with centrally mediated sympatholytic, sedative, and analgesic effects. This study evaluated: 1) pharmacokinetics of dexmedetomidine in plasma and cerebrospinal fluid (CSF) in surgical patients; 2) precision of a computer-controlled infusion protocol (CCIP) for dexmedetomidine during the immediate postoperative period; and 3) dexmedetomidine's sympatholytic effects during that period. Dexmedetomidine was infused postoperatively by CCIP for 60 min to eight women, targeting a plasma concentration (Cp) of 600 pg/mL. Before, during, and after infusion, blood was sampled to determine plasma concentrations of norepinephrine, epinephrine, and dexmedetomidine, and CSF was sampled to determine dexmedetomidine concentrations (C[CSF]). Heart rate and arterial blood pressure were measured continuously from 5 min before until 3 h after the end of infusion. During the infusion, Cp values generally exceeded the target value: median percent error averaged 21% and ranged from -2% to 74%; median absolute percent error averaged 23% and ranged from 4% to 74%. After infusion, C(CSF) was 4% +/- 1% of Cp. Because C(CSF) barely exceeded the assay's limit of quantitation, CSF pharmacokinetics were not determined. During the infusion, norepinephrine decreased from 2.1 +/- 0.8 to 0.7 +/- 0.3 nmol/L; epinephrine decreased from 0.7 +/- 0.5 to 0.2 +/- 0.2 nmol/L; heart rate decreased from 76 +/- 15 to 64 +/- 11 bpm; and systolic blood pressure decreased from 158 +/- 23 to 140 +/- 23 mm Hg. We conclude that infusion of dexmedetomidine by CCIP using published pharmacokinetic parameters overshoots target dexmedetomidine concentrations during the early postoperative period. Hemodynamic and catecholamine results suggest that dexmedetomidine attenuates sympathetic activity during the immediate postoperative period. IMPLICATIONS: We studied the pharmacokinetic and sympatholytic effects of dexmedetomidine during the immediate postoperative period and found that during this period, the published pharmacokinetic data slightly overshoot target plasma dexmedetomidine concentrations. We also found that heart rate, blood pressure, and plasma catecholamine concentrations decrease during dexmedetomidine infusion. PMID- 9356116 TI - The effects of premedication on inhaled induction of anesthesia with sevoflurane. AB - The effects of premedication with midazolam (M), fentanyl (F), or both (B) on induction of anesthesia via a mask with sevoflurane (S) were assessed in 24 healthy volunteers who participated on three occasions, receiving either intravenous (IV) F (2.4 microg/kg), M (36 microg/kg), or B (0.6 microg/kg F, 9 microg/kg M) 5 min before three vital capacity breaths of 8% S, 66% N2O, and O2. At loss of lid-lash reflex (LLR), ventilation was manually assisted until a randomly assigned time of administration was attained, at which time laryngoscopy and tracheal intubation were attempted. The effective times for 50% of subjects (ET50) to loss of LLR were 64 s for M and B and 54 s for F (P < 0.05). The ET50 to acceptable intubating conditions were 4.3, 3.1 and 2.5 min for F, M, and B, respectively. F resulted in more airway management difficulties than M or B. Heart rate was slightly increased before intubation in M. Heart rate increases after intubation were least in F, intermediate in B, and greatest in M. The time to achieve good intubating and airway conditions up to intubation was lowest with M or B. Anesthetic adjuvants did not improve the time to achieve loss of consciousness with anesthetic induction via the face mask with sevoflurane, but they significantly decreased the time to acceptable tracheal intubating conditions. IMPLICATIONS: Adults can be anesthetized with very few side effects by breathing themselves to sleep with sevoflurane. Giving patients small doses of sedatives intravenously before they inhale an anesthetic can improve the speed and quality of the process of falling asleep. PMID- 9356117 TI - Effects of inhaled nonimmobilizer, proconvulsant compounds on desflurane minimum alveolar anesthetic concentration in rats. AB - Anesthetics depress the central nervous system, whereas nonimmobilizers (previously called nonanesthetics) and transitional compounds having the same physical properties (e.g., solubility in lipid) do not produce anesthesia (nonimmobilizers) or are less potent anesthetics than might be predicted from their lipophilicity (transitional compounds). Potential explanations for the absent or decreased anesthetic effect of nonimmobilizer and transitional compounds include the theories that the nonimmobilizers are devoid of anesthetic effect and that transitional compounds have a decreased capacity to produce anesthesia; that the effects of these compounds are not apparent because the concentrations examined are too low; or that anesthesia, or lack thereof, results from a balance between depression and excitation (all nonimmobilizer and transitional compounds produce convulsions). To examine these issues further, we tested the effect of various multiples of the convulsive 50% effective dose (ED50) of three nonimmobilizers and one transitional compound on the minimum alveolar anesthetic concentration (MAC) of desflurane in rats. The nonimmobilizer 2,3-dichlorooctafluorobutane (NI-1), from 0.7 to 1.1 times its convulsive ED50, increased the MAC of desflurane by 14%-27%, but at 1.6 times its convulsive ED50 caused no change in MAC; the nonimmobilizer 1,2-dichlorohexafluorocyclobutane (NI 2) did not change MAC at concentrations up to its convulsant ED50, but it increased MAC by 25% and 36% at 1.3 and 1.7 times its convulsant ED50, respectively. The nonimmobilizer flurothyl (NI-3) decreased the MAC of desflurane by 20% +/- 6% (mean +/- SD) at 0.5 times its convulsant ED50, but it caused no change at higher partial pressures (up to 7.8 times its convulsant ED50), and the transitional compound CF3CCl2-O-CF2Cl (T-1) significantly decreased MAC by 16% +/ 7% at 0.8 times its convulsant ED50, but the 6%-8% decreases in MAC at 0.4 and 1.6 times its convulsant ED50 were not significant. Thus, neither nonimmobilizer nor transitional compounds produced a consistent dose-related effect on the MAC of desflurane, and any changes were small. These results suggest that the excitation produced by transitional compounds or nonimmobilizers does not explain their limited ability or inability to produce anesthesia. The data are consistent with a decreased anesthetic efficacy of transitional compounds and the lack of efficacy of nonimmobilizers. IMPLICATIONS: Inhaled compounds that do not cause anesthesia (nonimmobilizers) are used to test theories of anesthetic action. Their use presumes that a trivial explanation, such as cancelling stimulatory and depressant effects, does not explain the absence of anesthesia. The present results argue against such an explanation. PMID- 9356118 TI - Dose-related biochemical markers of renal injury after sevoflurane versus desflurane anesthesia in volunteers. AB - Sevoflurane (CH2F-O-CH[CF3]2) reacts with carbon dioxide absorbents to produce Compound A (CH2F-O-C[=CF2][CF3]). Because of concern about the potential nephrotoxicity of Compound A, the United States package label (but not that of several other countries) for sevoflurane recommends the use of fresh gas flow rates of 2 L/min or more. We previously demonstrated in humans that a 2-L/min flow rate delivery of 1.25 minimum alveolar anesthetic concentration (MAC) sevoflurane for 8 h can injure glomeruli (i.e., produce albuminuria) and proximal tubules (i.e., produce glucosuria and urinary excretion of alpha-glutathione-S transferase [alpha-GST]). The present report extends this investigation to fasting volunteers given 4 h (n = 9) or 2 h (n = 7) of 1.25 MAC sevoflurane versus desflurane at 2 L/min via a standard circle absorber anesthetic system (all subjects given both anesthetics). Markers of renal injury (urinary creatinine, albumin, glucose, alpha-GST, and blood urea nitrogen) did not reveal significant injury after anesthesia with desflurane. Sevoflurane degradation with a 2-L/min fresh gas inflow rate produced average inspired concentrations of Compound A of 40 +/- 4 ppm (mean +/- SD, 8-h exposure [data from previous study]), 42 +/- 2 ppm (4 h), and 40 +/- 5 ppm (2 h). Relative to desflurane, sevoflurane given for 4 h caused statistically significant transient injury to glomeruli (slightly increased urinary albumin and serum creatinine) and to proximal tubules (increased urinary alpha-GST). Other measures of injury did not differ significantly between anesthetics. Neither anesthetic given for 2 h at 1.25 MAC produced injury. We conclude that 1.25 MAC sevoflurane plus Compound A produces dose-related glomerular and tubular injury with a threshold between 80 and 168 ppm/h of exposure to Compound A. This threshold for renal injury in normal humans approximates that found previously in normal rats. IMPLICATIONS: Human (and rat) kidneys are injured by a reactive compound (Compound A) produced by degradation of the clinical inhaled anesthetic, sevoflurane. Injury increases with increasing duration of exposure to a given concentration of Compound A. The response to Compound A has several implications, as discussed in the article. PMID- 9356120 TI - Bilateral vocal cord paralysis after radical cystectomy in a patient with a history of bulbar polio. PMID- 9356121 TI - Difficult airway management with the intubating laryngeal mask. PMID- 9356119 TI - Quantitative differences in the production and toxicity of CF2=BrCl versus CH2F-O C(=CF2)(CF3) (compound A): the safety of halothane does not indicate the safety of sevoflurane. AB - Carbon dioxide absorbents degrade both halothane and sevoflurane to toxic unsaturated compounds (CF2=CBrCl and CH2F-O-C[=CF2][CF3] [i.e., Compound A], respectively). Given the long history of safe administration of halothane, comparable toxicities of these degradation products would imply a similar safety of sevoflurane. We therefore examined CF2=CBrCl in the context of four issues relevant to previous studies of the toxicity of Compound A: 1) reactivity of the degradation product in vitro; 2) rate of its production in vitro; 3) its in vivo toxicity; 4) importance of the beta-lyase pathway to the toxicity in vivo. We found the following. 1) CF2=CBrCl is less reactive than Compound A, degrading in human serum albumin at one-fifth the rate of Compound A. 2) Over a 3-h period of "anesthesia," a standard circle system containing Baralyme (Allied Healthcare Products, Inc., St. Louis, MO) produces 30 times as much Compound A from a minimum alveolar anesthetic concentration (MAC) concentration of sevoflurane as CF2=CBrCl from a MAC concentration of halothane; with soda lime, the difference is 60-fold. Correcting for differences in uptake of halothane versus sevoflurane decreases the differences to 20-40 times. 3) For a 3-h administration to rats, the partial pressure of Compound A causing minimal renal injury or necrosis of half the affected tubule cells exceeds the partial pressure of CF2=CBrCl causing minimal injury or necrosis of half the affected tubule cells by a factor of approximately 4-6. Thus, the ratio of production (Item 2 above) to the partial pressure causing injury with CF2=CBrCl is approximately a quarter of that ratio for Compound A. 4) Compounds that block the beta-lyase pathway either do not change (acivicin) or decrease (aminooxyacetic acid; AOAA) renal injury from CF2=CBrCl in rats, whereas these compounds increase (acivicin) or do not change (AOAA) injury from Compound A. We conclude that the safety of halothane cannot be used to support the safety of sevoflurane. IMPLICATIONS: Carbon dioxide absorbents degrade halothane and sevoflurane to unsaturated compounds nephrotoxic to rats. Relative to sevoflurane's degradation product, halothane's degradation product has less toxicity relative to production, less reactivity, and a different mechanism of injury. The clinical absence of halothane nephrotoxicity does not necessarily indicate a similar absence for sevoflurane. PMID- 9356122 TI - Pulmonary artery catheterization: high-tech or obsolete procedure? PMID- 9356123 TI - Emergence and extubation: a systematic approach. PMID- 9356124 TI - Anesthetic agents and platelet aggregation. PMID- 9356125 TI - Failure of a noninvasive automated blood pressure monitor to detect the blood pressure after repositioning of the patient's arm. PMID- 9356126 TI - Full disclosure in study design is essential. PMID- 9356127 TI - The flexible laryngeal mask as a nasal airway. PMID- 9356128 TI - Continuous stellate ganglion blockade revisited. PMID- 9356129 TI - Unbalancing the concept of the structured alarm systems for an anesthesia working area. PMID- 9356130 TI - Detection of tracheal stenosis by cineangiocardiography in infants with congenital heart disease. PMID- 9356131 TI - Management of difficult tracheal intubation with a video-optically modified schroeder intubation stylet. PMID- 9356132 TI - Is hydroxyethyl starch guilty or not? PMID- 9356133 TI - Spectrally constrained global analysis of fluorescence decays in biomembrane systems. AB - Dynamic and steady-state fluorescence spectroscopic properties of a dye probe measured in a multicomponent biological system are often required to be separated into the spectra and lifetimes of individual spectroscopically distinct species. The conventional method of obtaining decay-associated spectra fails when the lifetimes of the fluorophore in the membrane phase and in the aqueous phase are very close to each other. This paper describes a global analysis method which takes advantage of the known spectrum and lifetime of the dye in the aqueous phase. This method is used to identify the spectra for two fluorescent species (lifetimes, 0.84 and 1.97 ns) of the dye DODCI in EggPC vesicle membranes by keeping the spectrum and lifetime (0.68 ns) of the dye in the aqueous phase as fixed parameters. The structural identity of the two membrane-bound dye species was established by the effect of refractive index and/or viscosity of the aqueous medium on the lifetimes. PMID- 9356134 TI - Determination of trehalose by flow injection analysis using immobilized trehalase. AB - A new method for the determination of trehalose by flow injection analysis (FIA) is described. The basic principle is the hydrolysis of the disaccharide trehalose into its monomer d-glucose by trehalase, a periplasmic enzyme of Escherichia coli. d-glucose is quantified spectrophotometrically after reaction with hexokinase and glucose-6-phosphate dehydrogenase. Trehalase is prepared by osmotic shock from a recombinant E. coli strain and precipitated with ammonium sulfate. The enzyme is immobilized on VA-Epoxy Biosynth from Riedel-de-Haen. The immobilization rate is about 60%. The FIA signals show a nonlinear dependence on the trehalose concentration. The resulting curve corresponds to a second-order polynomial that serves as a calibration function for test samples. Immobilized trehalase was used during a period of 4 months without any loss of suitability. Several samples of fermentation broth were tested. The results are verified by HPLC. Within an interval of 2 to 10 g/L trehalose the recovery is about 100-120% with a precision of 7% (coefficient of variation). PMID- 9356135 TI - Evaluation of some fluorogenic substrates for continuous assay of aminopeptidase P. AB - Three potential fluorogenic substrates for assay of aminopeptidase P (AP-P) have been prepared and evaluated, using enzyme purified from porcine kidney. They are based on internal quenching of the synthetic, fluorescent amino acid (R, S)-2 amino-3-(7-methoxy4-coumaryl)propanoic acid ((R,S)-Amp) by a 2, 4dinitrophenyl (DNP) group. The compounds are X-Pro-Pro-(R, S)-Amp-NH2, where X is H-Lys(epsilon DNP), H-Orn(delta-DNP), or L-2-amino-3-(DNP)aminopropionic acid. The first two were found to be excellent substrates for AP-P, with respective Km values of 4.8 and 5.2 microM. An advantageous feature is that under the conditions of assay, using 4-mm2 cells, the substrates are without noticeable quenching effect on the fluorescence of Pro-Pro-(R,S)-Amp-NH2 (the product liberated by the action of AP P). At concentrations greater than about 30-50 microM, both substrates appear to inhibit the enzyme, but this has little practical consequence since assays can be carried out at substrate concentrations, giving up to approximately 80% of Vmax without this inhibitory effect being noticeable. The Lys derivative was found to be a very useful substrate for a continuous assay for AP-P and equally good in a discontinuous assay of multiple samples using microtiter plates. The racemic center at the Amp residue did not prevent total hydrolysis of the Lys derivative, suggesting that subsite specificity in AP-P does not extend as far as the P3' position. PMID- 9356136 TI - Determination of viridicatin in Penicillium cyclopium by capillary gas chromatography. AB - A sensitive gas chromatographic method for the determination of the quinoline alkaloid viridicatin, a fungal metabolite isolated from several Penicillium species and presumably a biogenetic precursor to naturally occurring diazepam like 1,4-benzo-diazepines, has been developed. Prior to conversion of viridicatin into a volatile trimethylsilyl derivative, a rapid sample clean-up was accomplished by solid-phase extraction from mould mycelium on a C18 cartridge. Using synthetical 6-bromo-viridicatin as an internal standard, the amount of viridicatin produced de novo in emerged grown cultures of Penicillium cyclopium strain SM 72 was calculated in correlation to the duration of mould cultivation. PMID- 9356138 TI - Determination of polyethylene glycol activated with cyanuric chloride in liposomes. AB - A simple method was developed for the determination of polyethylene glycol (PEG) 5000 activated with cyanuric chloride attached or adsorbed to the liposome membrane. The procedure is based in the measurement of the absorbance at 234 nm of the triazine ring of cyanuric chloride after disrupting the liposomal structure with CHCl3/MeOH (1/8, v/v). In this medium, the molar absorptivity of the molecule is the highest, and lipid does not interfere with the measurement. This procedure results in a convenient, sensitive, and rapid method for the determination of this kind of activated polyethylene glycol in liposomes. The minimal concentration of PEG measured is 50 microM, and so the method is sufficiently sensitive to quantify PEG 5000 in the minimal amounts used for protecting the liposomes from the action of serum. Values calculated by this method have been compared with those obtained by 1H NMR, and a good correlation between them has been obtained. PMID- 9356137 TI - Single-column high-performance liquid chromatographic-fluorescence detection of immature, mature, and senescent cross-links of collagen. AB - A high-performance liquid chromatographic-fluorescence detection method of reducible (immature) and nonreducible (mature and senescent) cross-links of collagen was established without the use of a radioisotope and preliminary fractionation step. This method used a gradient elution procedure of sodium citrate buffer containing 7% ethanol. The reducible cross-links (dihydroxylysinonorleucine, hydroxylysinonorleucine, and lysinonorleucine) and nonreducible cross-link (histidinohydroxylysinonorleucine) were detected by O phthalaldehyde derivatization with the postcolumn method, whereas other nonreducible cross-links (pyridinoline, deoxypyridinoline, and pentosidine) were detected by natural fluorescence. The linear ranges of contents of the O phthalaldehyde derivative cross-links and the natural fluorescent nonreducible cross-links were 20-600, 5-500 (pyridinoline, deoxypyridinoline), and 0.2-20 pmol (pentosidine), respectively. Tissue containing 1-2 mg dry wt of collagen was adequate for duplicate analyses of the reducible and nonreducible cross-links. An equivalent of 0.25 mg of hydrolyzed collagen could be analyzed by this HPLC system. Using this system, age-related changes in the cross-links of collagen from human connective tissues were also investigated. PMID- 9356139 TI - Determination of the relative affinities of antibody fragments expressed in Escherichia coli by enzyme-linked immunosorbent assay. AB - We have previously utilized an M13 phage expression system and codon-based mutagenesis to increase the avidity of the tumor-specific antibody, BR96, 65-fold (Yelton et al., 1995, J. Immunol. 155, 1994-2004). Mutants with improved affinity were identified by screening phage-expressed antibodies on a carcinoma cell line. In this study we describe a more broadly applicable assay which permits rapid and quantitative comparison of affinities of related antibodies produced in an M13 phage expression system. BR96 variants displaying a range of affinities were expressed as soluble antibody fragments (Fabs) in the periplasmic space of bacteria and isolated from small-scale cultures grown in a 96-well format, yielding between 142 ng and 1.06 microg of Fab. Although the small-scale cultures expressed variable levels of Fab, the lower quantities were sufficient to saturate microtiter plates coated with a limiting amount of anti-human Fab antibody, resulting in the capture of uniform quantities of the Fab variants. The relative affinities of the variants were then compared by assessing binding to biotinylated antigen followed by detection with streptavidin-alkaline phosphatase conjugates. This approach permitted the direct comparison of the relative affinities of large numbers of antibody variants in a single step without multiple antibody dilutions. The assay is readily adaptable for screening phage expressed antibody libraries against any biotinylated target and does not require purified antigen. For instance, the biotinylated BR96 antigen utilized in these studies was from a total cell extract prepared by labeling the surface of live tumor cells followed by detergent extraction. Thus, this approach may be applicable to the screening of antibody libraries against other cell surface antigens, such as transmembrane receptors, which are difficult to purify. PMID- 9356140 TI - A continuous coupled spectrophotometric assay for tyrosine aminotransferase activity with aromatic and other nonpolar amino acids. AB - A continuous assay for Escherichia coli tyrosine aminotransferase (TATase) that employs Lactobacillus delbrueckii ssp. bulgaricus hydroxyisocaproate dehydrogenase (HO-HxoDH) as a coupling enzyme is described. alpha-Keto acids, including those formed by TATase-catalyzed transamination of l-phenylalanine, l tyrosine, l-tryptophan, l-methionine, and l-leucine, are converted to the corresponding alpha-hydroxy acids by the auxiliary enzyme. The concomitant reduction of NADH by this enzyme can be followed as a decrease in absorbance at 340 nm. Importantly, HO-HxoDHcatalyzed reduction of alpha-ketoglutarate (alpha KG), a cosubstrate of TATase required to regenerate the pyridoxal-5'-phosphate cofactor of this enzyme from pyridoxamine-5'-phosphate, is a poor substrate and does not interfere with the assay. The kinetic parameters determined for the transamination of phenylalanine by TATase (kcat = 180 s-1, KM (L-Phe) = 0.56 mM, KM (alpha-KG) = 5 mM) with HO-HxoDH as a coupling enzyme are comparable to those reported in the literature, which were determined by direct monitoring of the formation of phenylpyruvate at 280 nm. This new assay offers the advantages of increased sensitivity and broad substrate specificity. PMID- 9356141 TI - Avoiding interferences from Good's buffers: A contiguous series of noncomplexing tertiary amine buffers covering the entire range of pH 3-11. AB - Of the 20 well-known buffers proposed by Good, all but 3 form metal ion complexes which can result in serious interferences, particularly in protein analyses. The structural features responsible for such complex formation have been identified. Based on a mechanistic analysis of the metal complexation process, it is proposed that tertiary amine compounds, having N-substituents which are ethyl or larger, are sterically inaccessible for initial bond formation with solvated metal ions in aqueous solution. Thus, in the absence of donor atoms on the alpha-, beta-, or gamma-carbons, metal complexation cannot proceed. The proposed noncomplexing compounds include Good's 3 noncomplexing buffers (Mes, Mops, Pipes) plus six related species. Mixed-mode acid dissociation constants (hydrogen ion in terms of activity, conjugate acid-base species in molar concentrations) have been determined for all compounds in their protonated form at 25 degrees C, mu = 0.10 M (NaNO3). The values for four compounds in this series are reported here for the first time: viz., N,N'-diethylpiperazine for which log Ka1m = 4.67 +/- 0. 03 and log Ka2m = 8.83 +/- 0.02; N,N,N',N'-tetraethylmethylenediamine for which log Ka1m < 1 and log Ka2m = 11.01 +/- 0.03; N,N'-diethyl-N,N'-bis(3 sulfopropyl)ethylenediamine for which log Ka1m = 5.75 +/- 0.03 and log Ka2m = 9.37 +/- 0.02; and piperazine-N,N'-bis(2-ethanesulfonic acid) (Pipes) for which log Ka1m = 2.81 +/- 0.01 (the value of log Ka2m having been previously reported). PMID- 9356142 TI - Correcting the circular dichroism spectra of peptides for contributions of absorbing side chains. AB - The aromatic and sulfur-containing side chains Trp, Tyr, Phe, Cys, and Met contribute to the CD spectra of peptides and proteins in the amide region, interfering with the analysis for secondary structure. We propose a method to correct the CD spectra of peptides undergoing the helix-coil transition for contributions due to absorbing side chains using singular value decomposition. The method uses the common basis vectors obtained from an analysis of the CD spectra of related peptides without the aromatic and sulfur-containing amino acids. The common basis vectors are fitted to a portion of the CD spectrum of the peptide being corrected, in the range that is unaffected by its sidechain contributions. Then the resulting coefficients from the fitting are used along with the common basis vectors to regenerate the entire corrected spectrum. The method is illustrated for the CD spectra of the peptide sequence acetyl-Y-VAXAK VAXAK-VAXAK-amide, where X is substituted with the 20 naturally occurring amino acids. This peptide model adopts a random-coil conformation in 2 mm sodium phosphate buffer, pH 5.5, and becomes an alpha helix in methanol/buffer solutions. The difference between the original and corrected spectra shows the contribution from the aromatic and sulfur-containing side chains. PMID- 9356143 TI - Characterization of insect cell lines: heteroduplex analysis employing a mitochondrial 16S ribosomal RNA gene fragment. AB - Routine cell line characterization procedures are not adequate for characterizing the cell lines of insect origin. Ribosomal RNA (rRNA) gene sequences and their comparisons have been used successfully for delineating species and phylogenetic analysis. Using similar principles, we have standardized a protocol for the confirmation of species identity of insect cell lines. The procedure includes PCR amplification of the mitochondrial 16S rRNA gene fragment from the cell line and larvae of known insect species and heteroduplex analysis to detect the sequence variation in the PCR-amplified rRNA gene fragments. If the PCR fragment of the cell lines yields a homoduplex with the larvae of known species, then the cell line is conspecific with the larvae. If the larvae and cell line are of two different species, then the analysis exhibits multiple bands of heteroduplexes. The technique also allows detection of cross-contamination of culture having two insect cell lines belonging to two different species. PMID- 9356144 TI - Analysis of interactions between huGATA-3 transcription factor and three GATA regulatory elements of HIV-1 long terminal repeat, by surface plasmon resonance. AB - Relative affinities of transcriptional regulatory elements for their respective factor have been essentially studied by bandshift analysis. Here we report a real time study of factor/DNA interactions using a surface plasmon resonance approach and further characterization of recovered proteins involved in this interaction. For this purpose, human GATA-3, either recombinant or in nuclear extracts, and three natural GATA elements of the HIV-1 long terminal repeat (sites 1, 2, and 3) were chosen, in which only site 2 is a noncanonical GATA site. Direct analysis of sensorgrams, with recombinant huGATA-3, allowed the comparison of association and dissociation profiles of the three DNA regions and their ranking according to their relative affinities. This result, confirmed by competitions with each GATA site, demonstrated the higher relative affinity (at least sevenfold) of site 3. Interactions between the canonical and unique GATA site 3 and nuclear extracts were also studied in real time and provided information on its association and dissociation rates for native huGATA-3. Finally, recovered protein was identified as genuine huGATA-3 by SDS-PAGE, Western blotting, and bandshift assays. PMID- 9356145 TI - Essential light chain exchange in smooth muscle myosin. AB - The exchange of 17-kDa essential light chain (LC17) in smooth muscle myosin was carried out by incubating myosin with a 10-fold molar excess of exogenously added LC17 over the corresponding endogenous light chain in the presence of trifluoperazine and 4.5 m ammonium chloride. Porcine aorta myosin contains two kinds of LC17 isoform, LC17nm and LC17gi, while chicken gizzard myosin contains only one kind of LC17 isoform. As LC17gi can be separated from LC17nm and gizzard LC17 by urea-gel electrophoresis, LC17nm in aorta myosin and LC17 in gizzard myosin were exchanged with LC17gi and LC17gi in aorta myosin was exchanged with LC17nm, and the degree of exchange was estimated by urea-gel electrophoresis. Under the optimal conditions (6 and 10 degrees C for aorta and gizzard myosin, respectively), nearly 90% of exchange, which is close to the theoretical value, was achieved for the former combinations, and a slightly lower exchange was obtained for the latter. The LC17-exchanged myosins contained stoichiometric amounts of the heavy and light chains and retained the original nature in the phosphorylation-dependent actin-activated ATPase activity, 6S-10S conformational transition, and filament assembly. PMID- 9356146 TI - The use of sequential high-performance liquid chromatography and capillary zone electrophoresis to separate the glycosylated peptides from recombinant tissue plasminogen activator to a detailed level of microheterogeneity. AB - The advantage of using the two-dimensional separation method was demonstrated by the separation of the complex tryptic peptides of recombinant human tissue plasminogen activator (rt-PA) glycoprotein. This method hybridizes two analytical methods where fractions containing glycopeptides are collected from reversed phase high-performance liquid chromatography (RP-HPLC) and separated by capillary zone electrophoresis (CZE). CZE readily resolved carbohydrate structural variants, based on sialic acid content and branching, on the same peptide. Nonglycosylated peptides incidentally collected in the same RP-HPLC fraction were well resolved from the glycopeptides. This combination of RP-HPLC and CZE was able to uniquely resolve all of the peptides and glycopeptide variants in rt-PA. Confirmation of the specific peaks in the CZE was made by matrix-assisted laser induced ionization time-of-flight mass spectrometry. The advantages and disadvantages of the existing methods for carbohydrate characterization in the biotech industry were compared. The advantage of this approach is to provide a simple extension of the existing tryptic digest protocols to include carbohydrate analysis to a detailed level of microheterogeneity. PMID- 9356147 TI - beta-Elimination of O-glycans from glycoproteins transferred to immobilon P membranes: method and some applications. AB - Selective beta-elimination of O-glycans from glycoproteins transferred from electrophoretic gels onto Immobilon P membranes is described. The experiments were performed with erythrocyte membrane proteins, in which glycophorins are the major poly-O-glycosylated components, and with lysates of human colon cancer cells CX-1.1. Lectins and monoclonal antibodies against peptidic, glycopeptidic, and carbohydrate epitopes were used to examine the effect of degradation. Experiments with erythrocyte membrane proteins showed that after heating the blots in 0.055 M NaOH for 16 h at 40 degrees C the O-glycans of glycophorins were undetectable, while N-glycans and peptidic epitopes of proteins were detected with unchanged or even increased intensity compared to untreated blots. The method was used to show that most protein-linked sialyl-Lea epitopes present on CX-1.1 cancer cells are located on O-glycosidic chains. Moreover, beta elimination on the blots allows examination of the dependence of peptidic epitopes on O-glycosylation. This was shown using monoclonal antibodies specific for blood group M- or N-related epitopes of glycophorin A (GPA). Most of these antibodies recognize glycopeptidic epitopes dependent on O-glycosylation and, therefore, they did not detect GPA on NaOH-treated blots. Some less frequent anti M antibodies cross-reacting with the rare GPA variant of Mg type are specific for a peptidic epitope which is unrelated to the MN blood group-specific amino acid sequence in unglycosylated peptides, but is recognized in GPA-M only in the glycosylated antigen. These antibodies, which showed specificity for GPA-M on untreated blots, detected GPA-M, GPA-N, and glycophorin B on NaOH-treated blots. PMID- 9356148 TI - Nonspecific amine immobilization of ligand can Be a potential source of error in BIAcore binding experiments and may reduce binding affinities. AB - The interaction of monovalent forms of NC41, an anti-viral neuraminidase antibody, and the antiidiotype antibody 11-1G10 has been used as a model system for BIAcore analysis to demonstrate the potential problems resulting from the nonspecific amine coupling procedure. To avoid complications due to antibody bivalency, monovalent Fab fragments and monomeric recombinant scFvs were used. When immobilized by amine coupling, the 11-1G10 anti-idiotype fragments were found to have an artificially reduced affinity for NC41 compared to the results obtained using site-directed immobilization via C-terminal thiol residue and from solution equilibrium measurements. The NC41 antibody fragments, on the other hand, were able to retain their 11-1G10 binding affinity when immobilized nonspecifically through free amine groups. These data, in combination with the known sequences of the two antibodies, suggested that nonspecific immobilization through one or more lysine residues close to or within the CDR2 region of the 11 1G10 VH domain was responsible for the reduced strength of the interaction with NC41. These results emphasize the need to use site-specific immobilization strategies when accurate kinetic measurements are required. PMID- 9356149 TI - Sensitivity enhancement of optical immunosensors with nanoparticles. AB - In recent years, several optical sensor techniques have been developed for the direct monitoring of biomolecular recognition processes at the surface of a sensor chip. Applications of these immunosensors for the determination of substances in serum could be demonstrated only for a few analytes due to the lack of sensitivity. Beside nonspecific binding of serum components to the sensor surface, the analytical sensitivity of these sensors is limited by the molecular weight of the analyte, so that smaller analyte molecules give only a moderate sensor response. In order to enhance the sensor signal, the use of mass labels, such as latex particles, was proposed in the literature. However, detection limits comparable to those of conventional ELISA techniques could not be realized so far. We demonstrate the optimization of a "nanoparticle enhanced immunosensor assay" for the detection of thyroid stimulating hormone, with respect to the particle coating, size, and nonspecific binding. The developed prototype assay requires a sample volume of 225 microL and has a measuring range up to 35 mIU/L. For the first time, we obtained a detection limit of 0.03 mIU/L (0.1 pm), which is fully competitive to conventional ELISA techniques. The assay allows serum samples to be measured with good precision and dilution linearity. The sensor can be reused several times and shows an excellent correlation to a commercial enzyme immunoassay. PMID- 9356150 TI - Determination of thioredoxin reductase activity in rat liver supernatant. PMID- 9356151 TI - Western blot of stained proteins from dried polyacrylamide gels. PMID- 9356152 TI - Calcium-modulated proteins change their immunoreactivity in the presence of Ca2+: a study of antibody recognition in a dot immunoassay for calmodulin, calcineurin (beta-subunit), and S100B. PMID- 9356153 TI - Separation of microsatellite fragments on small precast polyacrylamide gels and visualization by silver staining. PMID- 9356154 TI - Transformation of oral streptococci with single-stranded DNA plasmids and their compatibility. PMID- 9356155 TI - Bioaffinity chromatography in the 10 mM range of Kd. PMID- 9356156 TI - An alternative linker-mediated polymerase chain reaction method using a dideoxynucleotide to reduce amplification background. PMID- 9356157 TI - Evaluation of cell proliferation through a polymerase chain reaction-based cyclin A test. PMID- 9356158 TI - Application of formalin fixation to the purification of amyloid proteins. PMID- 9356159 TI - Environmental influences in hand preference: an African point of view. AB - In order to examine further the role of cultural and environmental factors in human manual preference, two surveys were undertaken in students from Ivory Coast and Sudan. In the first study (Abidjan, Ivory Coast) 382 secondary students, ages 12 to 22, answered a 20-item manual preference questionnaire. The observed frequency of left-hand preference was 7.9%, with very low left-hand use among the 18-22 age group (1%) and high among the 12-15 age group (14%). In the second study (Khartoum, Sudan) 759 undergraduates, ages 18 to 33, answered a 25-item questionnaire. The observed frequency of left manual preference was 5%. Subjects were also asked to indicate any pressure to change hand for writing, eating, or other manual activities and, in the second study, any upper limb injury which temporarily rendered the subject unable to use his (her) preferred hand. Report of an upper limb injury in the past was related to mixed (or inconsistent) hand preference. In both studies, the target activity against left-hand use was eating. These results show that cultural and environmental factors could change "natural" hand preference in three ways: (i) by changing the hand used for only one activity (e.g., eating), with no change for other familiar unimanual activities; (ii) by reducing the degree of hand preference; (iii) by changing the overall preferred hand, generally reducing the prevalence of left-handedness. The design of handedness studies should allow these possibilities to be distinguished. PMID- 9356160 TI - Menstrual cycle phase and mood effects on perceptual asymmetry. AB - Several studies have reported shifts in perceptual asymmetry during the menstrual cycle, but the potential confounding effect of mood changes has been largely ignored. In this study, 24 female subjects completed four visual laterality tasks and a mood questionnaire at three phases of the cycle. Results indicate no overall effect of cycle phase on any of the asymmetry or mood scores. However, results revealed significant associations between affect and perceptual asymmetry on a face perception task. Implications for mood effects on perceptual asymmetry and future research on cycle-related shifts in asymmetry are discussed. PMID- 9356161 TI - Profound developmental dyscalculia: evidence for a cardinal/ordinal skills acquisition device. AB - This article presents an analysis of a form of dysfunctional mathematical development. A child of normal intelligence was unable to acquire basic cardinal numerical skills, despite relatively intact ordinal number use. These findings provide evidence for an innate "cardinal/ordinal skills acquisition device" (COSAD). It is argued that if this COSAD is lacking, ordinal number use may be compensated for by linguistic logic and visual skills. Cardinal number skills, however, remain limited as these demand an innate internal representation of quantity, which cannot be compensated for. The findings are discussed in terms of different approaches to number development. PMID- 9356162 TI - Parsing schizophrenia with neurocognitive tests: evidence of stability and validity. AB - The stability and validity of a neurocognitive typology for schizophrenia were studied in 55 chronic patients who met DSMIII-R criteria for the illness. Subtypes were based on an earlier cluster analytic study by Heinrichs and Awad (1993) that utilized the following variables: IQ (WAIS-R), categories (Wisconsin Card Sorting Test), free recall intrusions (California Verbal Learning Test), and bilateral motor performance (Purdue Pegboard). Stability was examined by analyzing subtype assignment at the original assessment and 3 years later at follow-up. Stability over this interval was variable with an overall kappa of .45 and individual kappas from .12 to .66. Adjunct cognitive and clinical data gathered at follow-up provide evidence for the validity of several subtypes, especially in terms of their cognitive and functional differences. There was no evidence of symptom differences in this relatively asymptomatic medicated sample of patients. The results are discussed in terms of the possibility that several patterns of neurocognitive dysfunction may underlie schizophrenia, with implications for understanding the heterogeneity of the illness and its variable functional outcomes. PMID- 9356163 TI - Viewing strategies for simple and chimeric faces: an investigation of perceptual bias in normals and schizophrenic patients using visual scan paths. AB - Left hemi-face (LHF) perceptual bias of chimeric faces in normal right-handers is well-documented. We investigated mechanisms underlying this by measuring visual scan paths in right-handed normal controls (n = 9) and schizophrenics (n = 8) for simple, full-face photographs and schematic, happy-sad chimeric faces over 5 s. Normals viewed the left side/ LHF first, more so than the right of all stimuli. Schizophrenics viewed the LHF first more than the right of stimuli for which there was a LHF choice of predominant affect. Neither group demonstrated an overall LHF perceptual bias for the chimeric stimuli. Readjustment of the initial LHF bias in controls was probably a result of increased attention to stimulus detail with scanning, whereas the schizophrenics demonstrated difficulty in redirection of the initial focus of attention. The study highlights the role of visual scan paths as a marker of normal and abnormal attentional processes. PMID- 9356164 TI - Letter-case-specific priming in the right cerebral hemisphere with a form specific perceptual identification task. AB - In a form-specific perceptual identification task, subjects identify and write letter strings in the same letter case as they appear on a computer display. Letter-case-specific repetition priming was observed in this task when test items were presented directly to the right hemisphere, but not when they were presented directly to the left hemisphere, similar to results in previous word-stem completion experiments. This pattern of results was not obtained in a standard perceptual identification task. Results indicate that a specific visual-form subsystem, but not an abstract visual-form subsystem, operates more effectively in the right hemisphere than in the left, and task demands greatly affect which subsystems are recruited in different priming tests. PMID- 9356165 TI - Handedness and P300 from auditory stimuli. AB - The P3(00) event-related potential (ERP) was elicited in 20 left- and 20 right handed normal young adult male subjects using a simple auditory stimulus discrimination task. P3 amplitude from the target stimuli was larger at anterior electrode sites for left- compared to right-handed subjects. P3 latency from the standard stimuli was shorter for left- compared to right-handers. The N1, P2, and N2 components generally demonstrated similar handedness effects. The relationship of ERP amplitude and handedness to anatomical variables and cognitive factors is discussed. PMID- 9356166 TI - Distinct processes in line bisection according to severity of left unilateral spatial neglect. AB - We investigated the line bisection performances in 24 patients with left unilateral spatial neglect. They bisected lines of two lengths in three positions relative to the sagittal midplane of the body. The results showed that in the mild or moderate neglect patients, length and spatial locations of the lines affected the placement of the subjective midpoint. In the severe neglect patients, however, length had little effect on their performances, and location of the right endpoint in the egocentric space mainly determined the subjective midpoint. The line bisection process of the severe neglect patients was not only quantitatively but also qualitatively different from that of the mild or moderate cases. PMID- 9356167 TI - Onset of CNTFRalpha expression and signal transduction during neurogenesis in chick sensory dorsal root ganglia. AB - The expression of ciliary neurotrophic factor receptor alpha (CNTFRalpha) was investigated in the developing chick dorsal root ganglion (DRG) using affinity purified anti-CNTFRalpha antibodies. At thoracic levels, CNTFRalpha immunoreactivity (CNTFRalpha-IR) was first observed at stage 19 (E3) in cells with neuronal morphology. CNTFRalpha-IR is restricted to the neuronal lineage in the DRG throughout development. CNTFRalpha expression precedes that of neuron specific beta tubulin, Hu antigen, and Q211 antigen, which are markers expressed in developing sensory neurons. [3H]Thymidine-labeling studies showed the onset of CNTFRalpha expression during terminal mitosis of sensory neuron precursors, making CNTFRalpha the earliest known neuronal marker in the DRG. CNTFRalpha mediated signal transduction was demonstrated in E7 and E11 DRG neuron cultures by CNTF-induced STAT3 phosphorylation. Although low ligand concentrations (5 pM) elicit STAT3 phosphorylation in E7 and E11 DRG neurons, a survival response is only observed in neurons from E11 DRG. This implicates a complex readout mechanism downstream of STAT3 phosphorylation leading to different cellular responses that depend on the age of the DRG neuron. These results argue against a role of CNTFRalpha ligands in the control of early neuron survival but are compatible with other functions in neurogenesis and sensory neuron development. PMID- 9356168 TI - Graded and lamina-specific distributions of ligands of EphB receptor tyrosine kinases in the developing retinotectal system. AB - Molecular gradients have been postulated to control the topographic mapping of retinal axons in their central targets. Based initially on their expression patterns, and more recently on functional studies, members of the EphA subfamily of receptor tyrosine kinases and their ephrin-A ligands have been implicated in the guidance of retinal axons along the anterior-posterior axis of the chick optic tectum. The report that a receptor of the EphB subfamily, EphB2/Cek5/Nuk/Sek3, is expressed in a high ventral to low dorsal gradient in the developing chick retina and is present on ganglion cell axons suggests that it may be involved in the mapping of retinal axons along the corresponding dorsal ventral axis of the tectum. To address this issue, we have determined the expression and distribution of ephrin-B1/LERK-2/Cek5-L and ephrin-B2/LERK-5/Htk L/ELF-2, ligands for EphB2, in the developing chick retinotectal system using riboprobes, immunocytochemistry, and receptor affinity probes. Both ephrin-B1 and ephrin-B2 transcripts are expressed in a high dorsal to low ventral gradient in the developing retina, complementary to the distribution of EphB2. Ephrin-B1 and ephrin-B2 proteins are predominantly found in the developing plexiform layers, suggesting a role in the development of intraretinal connections. Neither protein is detected on ganglion cell axons. In tectum, ephrin-B1 transcripts are expressed in a high dorsal to low ventral gradient in the neuroepithelium and the protein is present along the processes of radial glia and is concentrated at their endfeet in the stratum opticum, at the time retinal axons are growing through it. This distribution of ephrin-B1 suggests that it influences retinal axon mapping along the dorsal-ventral tectal axis and may also be involved in intratectal development. In contrast, ephrin-B2 transcripts and protein are localized to the deeper retinorecipient laminae in the tectum at the time retinal axons begin to arborize in them, suggesting that this ligand may influence the laminar patterning of retinal axon terminations. PMID- 9356169 TI - Defects of the chorioallantoic placenta in mouse RXRalpha null fetuses. AB - The active derivatives of vitamin A (the retinoids) play important and multiple roles in mammalian development and homeostasis. We have previously shown that specific retinoic acid receptors are expressed in the chorioallantoic placenta of the mouse and that among these, RXRalpha is strongly expressed in the developing labyrinthine zone (Sapin, V., Ward, S. J., Bronner, S., Chambon, P., Dolle, P., Dev. Dyn. 208, 199-210, 1997). Here, we show that mouse fetuses with a targeted disruption of the RXRalpha gene develop defects of the chorioallantoic placenta. Both morphological abnormalities and alterations in the expression of molecular markers were found, mostly confined to the labyrinthine zone of placentas from mid-late gestation mutants. This region exhibited edema, abnormal stasis of maternal blood, and signs of disruption of the endothelial layer of fetal vessels. We also detected a reduction in the number of lipid droplets in the trophoblastic layer and abnormal fibrin deposits in the junctional zone of the mutant placentas. These abnormalities most probably result in an impairment of the functional capacities of exchange between the maternal and fetal circulations in the mutant placentas. Thus, placental defects could represent an extraembryonic cause of lethality for RXRalpha null mutant fetuses, in addition to the previously described embryonic cardiac defects. PMID- 9356170 TI - Separate elements in the 3' untranslated region of the mouse protamine 1 mRNA regulate translational repression and activation during murine spermatogenesis. AB - The mouse protamine mRNAs, Prm-1 and Prm-2, are translationally repressed for several days during male germ cell differentiation. The translational delay of mouse Prm-1 mRNA has previously been shown to be dependent upon cis-acting elements that reside in the last 62 nucleotides of the Prm-1 3' untranslated region (3' UTR). We have previously identified a 48/50-kDa protein that binds the 3' UTRs of both Prm-1 and Prm-2 mRNAs in a sequence-specific manner, is present in cytoplasmic fractions of postmeiotic round spermatids where the protamine mRNAs are translationally silent, and is markedly reduced in elongated spermatids where the protamine mRNAs become activated for translation. Surprisingly, the binding site for this activity maps to a region of the Prm-1 3' UTR not contained within the functional 62 nucleotides described above. In this report we show that the binding site for the 48/50-kDa protein can also delay translation of a reporter RNA in vivo, suggesting that the 48/50-kDa protein can repress the translation of Prm-1 mRNA during murine spermatogenesis. This observation proves that two separate regions of the Prm-1 3' UTR are sufficient to repress Prm-1 translation. In addition, immunocytochemistry and polysome analysis have revealed that this transgenic reporter mRNA fails to undergo proper translational activation. These results suggest that an additional region of the Prm-1 3' UTR is required for proper translational activation and that Prm-1 translational repression elements can be separated from those involved in translational activation. PMID- 9356171 TI - The intracellular calcium increase at fertilization in Urechis caupo oocytes: activation without waves. AB - The intracellular Ca2+ (Cai) increase at fertilization of the marine worm Urechis caupo (Echiura) was studied with conventional and confocal epifluorescence microscopy in oocytes microinjected with calcium green dextran or dually labeled with the calcium-insensitive dye tetramethylrhodamine dextran. Calcium green fluorescence was also measured with a photomultiplier system while the oocyte membrane potential was recorded and manipulated. The results show that Cai rises simultaneously around the oocyte cortex and peaks slightly later in the nucleoplasm. The Cai rise coincides with the initiation of the fertilization potential and we conclude that it is due primarily to external Ca2+ entering through the voltage-gated Ca2+ action potential channels that open during the fertilization potential because: (1) current clamping the oocyte membrane potential to positive values in the absence of sperm produces a similar Cai increase, (2) external Ca2+ is required, (3) and the confocal images are consistent with this mechanism. External application of sperm acrosomal peptide (P23) also caused a Cai increase that was inhibited in the presence of CoCl2. Cai and pHi (measured with BCECF dextran) were manipulated in experiments employing microinjection of BAPTA (to chelate Cai), external application of NH4Cl (to increase pHi) and CoCl2 (to block Ca2+ channels), and fertilization of eggs in pH 7 seawater (Cai increase without pHi increase). The results showed that increases in both Cai and pHi are required for GVBD; neither alone is sufficient. However, although nuclear and cytoplasmic Ca2+ levels tended to parallel each other in oocytes fertilized at pH 7, and during the initial Cai response in oocytes fertilized at pH 8, there was a disproportionate fluorescence increase in the nucleoplasm of the latter prior to GVBD which could not be explained by any artifact we tested, suggesting there may be a selective increase in nuclear Ca2+ associated with GVBD. Finally, electrophysiological experiments with BAPTA injected oocytes showed that the opening of the fertilization potential Na+ channels was Ca2+-independent, (although they did not close at the normal time). These and earlier results suggest that Urechis sperm may activate oocytes by interacting directly with the Na+ channels or associated receptors. PMID- 9356172 TI - Dorsal determinants in the Xenopus egg are firmly associated with the vegetal cortex and behave like activators of the Wnt pathway. AB - The Xenopus egg contains maternal dorsal determinants that are specifically located at the vegetal cortex. To study physical and functional properties of the dorsal determinants, we took advantage of the animal-vegetal reversed embryo. The animal-vegetal reversed embryo is produced by inversion of the fertilized egg, which results in formation of ectoderm and endoderm from the unpigmented and the pigmented halves, respectively [Neff et al. (1983). Dev. Biol. 97, 103-112; Black and Gerhart (1985). Dev. Biol. 108, 310-324]. We demonstrated by cytoplasmic transplantation that the dorsal activity was specifically localized to the unpigmented cortical cytoplasm of the inverted egg, which is segregated into the future ectodermal lineage. This result suggests that the dorsal determinants are associated with the unpigmented cortex and are not dislodged by the inversion. In addition, we found that two vegetally localized transcripts, Xcat2 and Vg1 mRNAs, were present in the reversed animal pole of the inverted egg, suggesting their association with the unpigmented cortex. In order to compare the dorsal determinant activity with known dorsalizing molecules, we examined the expression pattern of Xnr3 and Siamois in the reversed embryo because these two genes are activated by the Wnt-pathway activators (Xwnt-8, beta-catenin, etc.) but not by other dorsalizing molecules (noggin, BVg1, etc.). Animal cap of the reversed embryo, which received the unpigmented cortex of the egg, expressed Xnr3 and Siamois. However, Mix.1, a marker expressed in endoderm and mesoderm in the normal embryo in response to mesodermal inducers, was not detected in the animal cap of the reversed embryo. In addition, we found that beta-catenin protein accumulated in nuclei of unpigmented animal pole cells of the reversed embryo. These results suggest that the maternal dorsal determinants behave more similarly to the Wnt-pathway activators than noggin or BVg1. PMID- 9356173 TI - The exocytosis regulatory proteins syntaxin and VAMP are shed from sea urchin sperm during the acrosome reaction. AB - Syntaxin is a cytoplasmically oriented plasma membrane protein and VAMP (vesicle associated membrane protein; synaptobrevin) is a protein associated with the secretory vesicle membrane. These two proteins form part of a complex which is thought to mediate the fusion of plasma and vesicle membranes during exocytosis. This paper reports the identification of syntaxin and VAMP homologues in sea urchin sperm. During fertilization, sea urchin sperm release the contents of a single vesicle, the acrosomal vesicle, exposing the membrane destined to fuse with the egg. During acrosomal exocytosis, the plasma membrane over the acrosomal vesicle fuses at multiple points with the acrosomal membrane (vesiculation) and syntaxin and VAMP are shed with the resulting membrane vesicles. Sea urchin sperm syntaxin and VAMP are associated in a complex as detected by immunoprecipitation. Following acrosomal exocytosis, syntaxin and VAMP cosediment to denser fractions on sucrose gradients showing that they have undergone associative changes during or after the acrosome reaction. Syntaxin and VAMP localization and loss during acrosomal exocytosis support a role for these proteins in regulating the acrosome reaction. PMID- 9356174 TI - The differential effects of 12-O-tetradecanoylphorbol-13-acetate on the gap junctions and connexins of the developing mammalian lens. AB - Epithelial cells in primary ovine lens cultures express the gap junction proteins connexin43 (Cx43) and connexin49 (Cx49; a.k.a. MP70), a homologue of mouse connexin50. In contrast, lens cultures of differentiated, fiber-like cells (termed lentoid cells) express Cx49 and connexin46 (Cx46), but not Cx43. To investigate the regulation of lens cell gap junctions by protein kinase C (PKC), differentiating lens cultures were treated with the PKC activator 12-O tetradecanoylphorbol-13-acetate (beta-TPA). Within 10 min, beta-TPA significantly inhibited the transfer of Lucifer Yellow dye between epithelial, but not lentoid, cells. This inhibition was correlated with the phosphorylation of Cx43 and was followed by the gradual disappearance of Cx43 from cell interfaces. The protein kinase inhibitor staurosporine prevented Cx43 phosphorylation and the loss of Cx43 from intercellular junctions. Following treatment of cultures with beta-TPA for 2-6 hr, Cx49 disappeared from epithelial cell interfaces, and by 24 hr of beta-TPA treatment, levels of Cx49 detected on immunoblots of purified epithelial membrane fractions had also diminished significantly. The beta-TPA-induced loss of Cx49 both from regions of epithelial cell contact and from isolated membranes was correlated with the disappearance of Cx49 mRNA. In contrast to the epithelial connexins, the lentoid connexins Cx49 and Cx46 were unaffected by even extended beta-TPA treatment. In spite of lentoid dye transfer being refractory to beta TPA, significant levels of PKC-alpha (a beta-TPA-sensitive isoform) were detected in the lentoid cell. The response of lens gap junctions to beta-TPA depends upon the stage of differentiation and the complement of connexins expressed. The contrasting effects of beta-TPA on Cx43 and Cx49 in lens epithelial cells indicate a fundamental difference in the regulation of these connexin proteins in the developing mammalian lens. PMID- 9356175 TI - GFP-moesin illuminates actin cytoskeleton dynamics in living tissue and demonstrates cell shape changes during morphogenesis in Drosophila. AB - Moesin, ezrin, and radixin (MER) are components of the cortical actin cytoskeleton and membrane processes such as filopodia and microvilli. Their C terminal tails contain an extended region that is predicted to be helical, an actin binding domain, and a region(s) that participates in self-association. We engineered an in vivo fluorescent actin binding protein (GFP-moe) by joining sequences that encode the jellyfish green fluorescent protein (GFP) to sequences that encode the C-terminal end of the sole Drosophila MER homolog, moesin [Moesin like gene product, referred to previously as the D17 MER-like protein; Edwards et al., 1994, Proc. Natl. Acad. Sci. USA 91, 4589], and Dmoesin [McCartney and Fehon, 1996, J. Cell Biol. 133, 843]. Transgenic flies expressing this fusion protein under control of the hsp70 promoter were generated and used for analysis of cell shape changes during morphogenesis of various developmental stages and tissues. Following heat shock, high levels of stable fusion protein are produced by all somatic tissues. GFP-moe localizes to the cortical actin cytoskeleton, providing a strong in vivo marker for cell shape and pattern during epithelial morphogenesis. The protein also becomes highly enriched in pseudopods, microvilli, axons, denticles, the border cell process, and other membrane projections, potentially by binding to endogenous moesin as well as actin. We show that GFP-moe can be used to examine the development and behavior of these dynamic structures in live specimens. We observe a bright green fluorescent, presumably actin-rich, polar cell proboscis that inserts itself into the forming micropyle and appears to maintain an opening for sperm passage around which the chorion is formed. We also confirm the existence of an actin-rich purse string at the leading edge of the lateral epidermis and provide a dynamic analysis of its behavior as it migrates during dorsal closure. Observations of embryos, larvae, and pupae show that GFP-moe is also useful for labeling the developing nervous system and will be a good general marker of dynamic cell behavior during morphogenesis in live tissues and demonstrate that fusion of a subcellular localization signal to GFP greatly increases its utility as a cell marker. PMID- 9356177 TI - Evidence for distinct serine protease activities with a potential role in processing the sperm protein fertilin. AB - The guinea pig sperm protein fertilin (previously termed PH-30) plays an important role in sperm-egg fusion, and was the first recognized membrane anchored metalloprotease/disintegrin protein. Fertilin is a heterodimeric glycoprotein which undergoes at least two distinct proteolytic processing steps. Fertilin alpha is processed first, in the testis, whereas fertilin beta is processed separately during sperm maturation in the epididymis. The final processing of fertilin beta occurs immediately adjacent to its predicted integrin ligand domain, and exposes an epitope recognized by a fusion blocking monoclonal antibody. Here, we demonstrate that one or more serine protease activities associated with testicular sperm can process fertilin beta in vitro in a fashion that closely mimics the processing pattern observed in vivo during epididymal sperm maturation. In contrast, several proteases that were added to testicular sperm did not mimic the pattern observed in vivo. These findings raise the intriguing possibility that a fertilin beta converting protease(s) active in vivo may originate from sperm, instead of from the epididymal epithelium. Further, we show that fertilin alpha is most likely processed intracellularly in the secretory pathway based on three observations: (i) only processed fertilin alpha, but not the precursor pro-alpha can be cell-surface biotinylated; (ii) some processed fertilin alpha is sensitive to endoglycosidase H, suggesting cleavage occurs prior to the medial Golgi apparatus; (iii) a reanalysis of the N-terminus of processed fertilin alpha showed that the proteolytic cleavage site is next to four arginine residues, a consensus sequence for intracellular subtilysin type pro-protein convertases. The N-terminal sequence analysis further showed that processed fertilin alpha contains an intact membrane anchored disintegrin domain, and not a truncated disintegrin domain as reported previously (Blobel, C. P., Wolfsberg, T. G., Turck, C. W., Myles, D. G., Primakoff, P., and White, J. M., Nature 356, 248-252, 1992). Proteolytic processing is thought to play an important role in regulating the function of fertilin, and the present study represents a first step toward a better understanding of protease activities involved in the maturation of fertilin, and potentially other sperm surface proteins. PMID- 9356176 TI - glide/gcm is expressed and required in the scavenger cell lineage. AB - Glial cell differentiation in Drosophila melanogaster requires the activity of glide/gcm (glial cell deficient/glial cell missing). The role of this gene is to direct the cell fate switch between neurons and glial cells by activating the glial developmental program in multipotent precursor cells of the nervous system. In this paper, we show that glide/gcm is also expressed and required in the lineage of hemocytes/macrophages, scavenger cells that phagocytose cells undergoing programmed cell death. In addition, we show that, as for glial cells, glide/gcm plays an instructive role in hemocyte differentiation. Interestingly, it has been shown that in the development of the fly adult nervous system the role of scavenger cells is played by glial cells. These data and our findings on the dual role of glide/gcm indicate that glial cells and hemocytes/macrophages are functionally and molecularly related. PMID- 9356178 TI - Analysis of the process of localization of fertilin to the sperm posterior head plasma membrane domain during sperm maturation in the epididymis. AB - Fertilin is a heterodimeric (subunits alpha and beta) sperm plasma membrane protein. Both subunits belong to the ADAM protein family of surface proteins that contain a disintegrin and a metalloprotease domain. Fertilin functions in sperm egg fusion by binding the sperm to the egg plasma membrane via a binding site in the disintegrin domain of fertilin beta. On testicular sperm of guinea pig, fertilin is distributed on the plasma membrane over the entire sperm head, but is found only on the posterior head once sperm have passed through the epididymis. This redistribution of fertilin to the posterior head can be partially mimicked in vitro if testicular sperm are briefly treated with trypsin. In this study we used immunofluorescence and digital image analysis to analyze how fertilin becomes restricted to the posterior head. We found that fertilin became restricted to the posterior head by migration of anterior head fertilin molecules into the posterior head domain. Comparison of immunofluorescence patterns and immunoblots of fertilin from seven regions of the epididymis showed a temporal correlation between the beginning of fertilin's migration to the posterior head and the proteolytic processing of the full-length fertilin beta precursor (the 85 kDa pro-beta form) to a 75-kDa intermediate, pro-beta*. Completion of the migration coincided with the further cleavage of pro-beta* to the 25- to 28-kDa mature form. Our data suggest that the cleavage of fertilin pro-beta to pro-beta* may initiate fertilin's migration into the posterior head domain and, after localization to that membrane domain, pro-beta* is cleaved to mature beta. We also report evidence that a common mechanism may be used to change the localization pattern of other sperm surface molecules. Other surface proteins were shown to become localized to either the posterior or the anterior head membrane domains on sperm at the same time fertilin became localized to the posterior head. These restrictions of surface protein localizations were also shown to immediately precede the development of the sperm's ability to swim and undergo the acrosome reaction, and thus redistribution of surface proteins may be necessary before sperm become functional. PMID- 9356180 TI - ANNOUNCEMENT PMID- 9356181 TI - The Effect of Achievement Goals: Does Level of Perceived Academic Competence Make a Difference? AB - Researchers using a goal orientation framework have hypothesized that learning goals are associated with adaptive patterns of behavior, regardless of the level of perceived ability. In contrast, perceived ability is hypothesized to moderate the relation between performance goals and patterns of adaptive or maladaptive behavior. We examined this hypothesis in two samples of seventh grade middle school students, focusing on the math domain in one sample and on the English domain in the other. Using two different statistical methods, median split and multiple regression, we found only little support for the role of perceived competence as a moderator between performance goals and patterns of behavior. Contrary to what has been suggested, we found some evidence that perceived competence moderated the relation between learning goals and behavior. Implications of these findings for recent efforts to use goal theory to reform classrooms and schools are discussed. Copyright 1997Academic Press PMID- 9356179 TI - A role for WNT proteins in induction of dermomyotome. AB - Dorsoventral patterning of somites into sclerotome and dermomyotome involves antagonistic actions of ventralizing and dorsalizing signals originating from tissues surrounding the somites. The notochord and the floor plate of the neural tube provide a ventralizing signal(s) directing sclerotome development, whereas the surface ectoderm and dorsal neural tube provide a dorsalizing signal(s) directing dermomyotome development. Evidence has been provided that Sonic Hedgehog mediates the ventralizing effects of notochord and floor plate, but the dorsalizing signal(s) that patterns the dermomyotome has not been identified. The documented expression of Wnt1 and Wnt3a in the dorsal neural tube and of Wnt4 and Wnt6 in the surface ectoderm at the time of dermomyotome specification prompted us to investigate the involvement of WNT proteins in patterning the dermomyotome. Here we show that tissue culture cells expressing these WNT family members can maintain and induce dermomyotome marker expression in presomitic mesoderm explants, supporting the hypothesis that WNT proteins mediate the dorsalizing effects of the surface ectoderm and dorsal neural tube on somites. PMID- 9356182 TI - Situational Interest in Literary Text AB - This study examined relationships among text characteristics, situational interest, two measures of text understanding, and personal responses when reading a literary text. A factor analysis of ratings made after reading revealed six interrelated text characteristics. Of these, suspense, coherence and thematic complexity explained 54% of the variance in interest. Additional analyses found that situational interest was unrelated to a multiple choice test of main ideas; but was related to personal responses and holistic interpretations of the text. These results suggest that multiple aspects of literary texts are interesting to readers, and that interest is related to personal engagement variables, even when it is not related to the comprehension of main ideas. Copyright 1997Academic Press PMID- 9356183 TI - Validation of the Metacomprehension Scale AB - Evidence for the factorial, convergent and discriminant, and criterion-related validity of the Metacomprehension Scale (MCS) was examined in a sample of 237 young adults. The instrument was factorially heterogeneous but exhibited homogeneity within each of the seven subscales. Evidence for the convergent and discriminant validity of the MCS was examined by correlating the subscales from the MCS with subscales from metacognitive questionnaires measuring similar constructs from related domains. In general, correlations within constructs were larger than correlations between constructs, providing preliminary evidence of the convergent and discriminant validity of the MCS. The criterion-related validity of the MCS relative to other metacognitive measures was examined by using the metacognitive measures and the MCS to predict comprehension performance. The MCS predicted performance better than the other measures of metacognition and accounted for variance in performance not accounted for by the other measures. These results show promise for the value of self-assessments of metacomprehension. Copyright 1997Academic Press PMID- 9356184 TI - An Inquiry into the Spontaneous Transfer of Problem-Solving Skill AB - Problem solving, by definition, involves achieving new understanding in unfamiliar contexts, and is critical to all aspects of life, especially in the educational and scientific arenas. Students learn from many experiences to develop a repertoire of abilities, including the use of logic, which enable them to spontaneously transfer their problem-solving skill to unfamiliar situations. The purpose of this study is to explore the minimum conditions necessary to facilitate the spontaneous transfer of problem solving skill in an unfamiliar context. One hundred and seventy-five subjects were presented with logically identical problems based on the Wason selection task, which differed only in the degree to which a familiar schema could be invoked to help solve the problem. In the pretest stage, only 10.5% of subjects could solve the selection problem in an unfamiliar context, whereas 57.3% could solve it in one that was familiar. The effect of three interventions, prior exposure to a familiar scenario, repeat opportunities on like problems, and process-oriented feedback, on selection task performance in an unfamiliar context was assessed in a posttest stage. Overall, none of the interventions were effective, indicating that the minimum threshold for spontaneous transfer may be above the level of intervention included in this study. Schema theory, implications for instruction, and directions for future research are discussed. Copyright 1997Academic Press PMID- 9356185 TI - Training in Place-Value Concepts Improves Children's Addition Skills AB - The present study examined whether children's variation in arithmetic performance was related to differences in their understanding of place-value. Training in place-value concepts was provided to a group of Chinese children who were poor in arithmetic. Their performance before and after the training was compared to that of the children in two control groups. The results showed that there were reliable connections between place-value understanding and addition and subtraction skills. Furthermore, training in place-value concepts was found to be effective in enhancing the children's place-value understanding and addition skills. Implications for instructions in arithmetic were discussed. Copyright 1997Academic Press PMID- 9356186 TI - Metamemory Assessment and Memory Behavior in a Simulated Memory Professional Task AB - This study investigated metamemory knowledge related to a professional task and the relationships between metamemory knowledge and memory performance in a simulated professional task, which was a beverage-service job with memory constraints changes. Metamemory knowledge was assessed by interviewing student waiters about hypothetical recall tasks concerning lists of beverages. They then carried out a simulated beverage-service task, including a first paired-associate recall (beverage-customer), then a global recall (order to the bartender), followed by a second paired-associate recall (beverage-customer). Memory constraints were manipulated with table size and perceptive cues. Results revealed that in metamemory knowledge, task-strategy, and strategy were the only variables that were related. Metamemory knowledge produced an effect on all memory performance, whatever the constraints were. The implications of these findings for professional training are discussed in terms of strategy instruction for enhancing professional performance when memory demands change in the work environment. Copyright 1997Academic Press PMID- 9356187 TI - Recall of Descriptive Information: The Roles of Presentation Format, Annotation Strategy, and Individual Differences AB - The benefits of using a comprehensive annotation strategy (employing underlining/circling, making connections, asking questions, and making comments) with knowledge maps (spatial/verbal arrays) and traditional, linear text to improve free recall scores for learners with individual differences in vocabulary and comprehension ability were examined. Types and frequencies of annotations generated were also examined for each stimulus format condition. Multiple regression analyses indicate that the frequency of use of two component annotation strategies, asking questions and making connections, were significant predictors of recall scores, while frequency of underlining/circling and generating elaborations failed to predict recall scores. Text users generated more underlining/circling, while knowledge map users generated more connections between ideas, suggesting that knowledge maps may facilitate the application of more productive annotation strategies. Also examined were the interrelationships between vocabulary ability, comprehension ability, and free recall scores. Copyright 1997Academic Press PMID- 9356188 TI - Seroimmunological characteristics of Korean workers exposed to toluene diisocyanate. AB - Since type I allergy caused by specific IgE antibodies may play principal roles and IgG antibody-mediated reactions have been thought to be involved in some parts of the pathogenesis, this study was performed to investigate the role of IgE- or IgG-mediated hypersensitivity reactions in development of toluene diisocyanate (TDI) asthma in Korean workers. For 81 TDI spray painters, self administrative questionnaires and direct interviews on respiratory symptoms, chest auscultation, and measurements of forced vital capacity (FVC) and forced expiratory volume in 1 sec (FEV1.0) were performed. The TDI concentration in their working environments was measured. Levels of serum IgE and IgG specific to TDI were estimated by radioallergosorbent test (RAST) and ELISA using p-tolyl isocyanate-human serum albumin (TMI-HSA) as the antigen. When sputum, cough, and dyspnea aggravated by work or wheezing existed, when FVC or FEV1.0 was less than 80% of the normal reference value, or when IgE RAST for TDI was positive, the peak expiratory flow rate (PEFR) was recorded four times per day for over 2 weeks. If decrease of PEFR was over 20% of baseline PEFR and changing pattern of PEFR was closely related to workshift in time, then a diagnosis of TDI asthma was made. Changing patterns of PEFR of 8 (9.9%) workers corresponded to the diagnostic criteria of TDI-related occupational asthma. Levels of the specific IgE were increased in 9 (11.1%) of the 81 subject workers and in 3 (37.5%) of the 8 PEFR-positive workers. Levels of the specific IgG were increased in 9 (11.1%) workers, and in only 1 (12.5%) of the asthmatics sensitive to TDI. Neither elevated TDI-specific IgE levels nor PEFR test positivities were associated with increased IgG levels. The mean titer of the PEFR-test-positive workers was slightly lower than that of the PEFR-negative workers and that of the IgE RAST positive workers lower than that of the test-negative workers, but there was no statistical significance. These results suggest that IgG is not deeply involved in the pathogenesis of TDI-induced occupational asthma in Korean workers. PMID- 9356189 TI - Early pulmonary cytokine responses to zinc oxide fume inhalation. AB - Zinc oxide inhalation causes metal fume fever, a flu-like syndrome common among welders. Proinflammatory pulmonary cytokines play a role in mediating this occupational illness. The goal of this investigation was to characterize early pulmonary cytokine responses after experimental human exposure to inhaled purified zinc oxide fume. We quantified bronchoalveolar lavage (BAL) cytokine concentrations in 15 healthy volunteers 3 hr after inhalation of zinc oxide fume. We compared postexposure cytokine responses with postsham exposure responses in the same 15 subjects. We also compared cytokine responses with those of 14 "late follow-up" subjects previously studied by BAL 20 hr after zinc oxide fume exposure. Zinc oxide exposure was a statistically significant, dose-dependent predictor of increases in BAL TNF (mean exposure-sham difference +/- SE = 9.5 +/- 3.6 pg/mL, P = 0.02), IL-6 (mean exposure-sham difference +/- SE = 5.5 +/- 1.8 pg/mL, P = 0.009), and IL-8 (mean exposure-sham difference +/- SE = 64.1 +/- 23.9 pg/mL, P = 0.02). The TNF response was significantly greater at 3 hr follow-up compared with 20 hr follow-up, after adjusting for smoking status, zinc dose, and BAL macrophages (P = 0.004). Our findings provide evidence for a pulmonary inflammatory response 3 hr after inhalation of zinc oxide fume characterized by dose-dependent increases in BAL proinflammatory cytokine concentrations. These data indicate that TNF plays an important initial role in mediating metal fume fever. PMID- 9356190 TI - Tears while cooking: an indicator of indoor air pollution and related health effects in developing countries. AB - Indicators for cooking fuel pollution are needed to determine the extent of fuel related problems in developing countries and to assess the success of measures undertaken to reduce such problems. It is proposed that eye irritation in the form of tears or smarting eyes during cooking time [tears while cooking (TWC)] is a useful determinant of indoor air pollution from cooking-related sources. An analysis of data from three cities (Lusaka, Maputo, and Hanoi) showed that TWC is more prevalent in conditions of higher particulate pollution. Persons experiencing TWC were also found to have more respiratory symptoms. The prevalence of TWC provides a good indicator of groups that are at greater risk of health impairment due to indoor air pollution. Surveying for this condition is simple and nonintrusive, which makes it a useful screening indicator, though it cannot replace more thorough investigations in epidemiological studies. It can, however, be used in guiding interventions to the most needy groups. It can also be used to assess the success of measures introduced to reduce pollution hazards relating to the cooking environment, such as improved stoves, chimneys, and kitchen improvement. PMID- 9356192 TI - Mesothelioma incidence and community asbestos exposure. AB - This study evaluates the environmental, nonoccupational component of mesothelioma incidence among persons living in Manville, Somerset County, New Jersey, the location of the largest asbestos manufacturing plant in North America. Prior to removal of occupational cases, residents of Manville had an average annual (1979 1990) mesothelioma rate of 636 male cases and 96 female cases per million population, about 25 times higher than average state rates. Somerset County had 143 diagnosed mesothelioma cases reported to the population-based. New Jersey State Cancer Registry from 1979 through 1990. Cases were removed from the analysis when their "usual employment" was reported as being at the asbestos plant, as evidenced through union lists or occupational information from either the Cancer Registry or mortality records. Standardized incidence ratios (SIRs) were computed for residents of Manville and Somerset County (less the Manville population) by sex. New Jersey mesothelioma rates less the Somerset County contribution, 1979-1990, were used to generate the expected number of cases. The SIRs for Manville males and females were respectively 10.1 [95% confidence interval (CI): 5.8-16.4] and 22.4 (95% CI: 9.7-44.2). Male and female Somerset County mesothelioma incidence rates were 1.9 (95% CI: 1.4-2.5) and 2.0 (95% CI: 1.0-3.6). This record-based approach demonstrates a strong relationship between past asbestos exposure from living in Manville and eventual development of mesothelioma. The use of methods in this study may be helpful in evaluating hazards of other known occupational carcinogens found in community settings. PMID- 9356191 TI - The assessment of biomarkers to detect nephrotoxicity using an integrated database. AB - Groups of industrial workers exposed to heavy metals (cadmium, mercury, and lead) or solvents were studied together with corresponding control groups. The cohorts were collected from several European centers (countries). Eighty-one measurements were carried out on urine, blood, and serum samples and the results of these analyses together with questionnaire information on each individual were entered into a central database using the relational database package Rbase. After the completion of the database construction phase, the data were exported in a format suitable for analysis by the statistical package SAS. The potential value of each test as an indicator of nephrotoxicity was then assessed. Rigorous exclusion criteria were applied which resulted in the elimination of some tests and samples from the dataset. The measurable contributions of smoking, gender, metal exposure, and site were either singly or in combination assessed by biomarkers for nephrotoxicity. The parameters measured included three urinary enzymes, six specific proteins, total protein, two extracellular matrix markers, four prostaglandins and anti-GBM antibodies, and beta 2-microglobulin in serum. The most sensitive renal tests included the urinary enzymes N-acetyl-beta-D glucosaminidase (NAG) and intestinal alkaline phosphatase (IAP), brush border antigens, and urinary low-molecular-weight proteins. Of the newer tests investigated the prostaglandins were the most promising. Different patterns of biomarker excretion were observed following exposure to lead, cadmium, or mercury. The dataset provides a unique repository of data which could provide the basis of an enlarging source of information on normal human reference ranges and on the effects of exposure to toxins and the use of biomarkers for monitoring nephrotoxicity. PMID- 9356193 TI - Zinc and copper levels in ribs of cadmium-exposed persons with special reference to osteomalacia. AB - Cadmium (Cd), zinc (Zn), copper (Cu), calcium (Ca), phosphorus (P), and magnesium (Mg) were determined in ribs obtained at autopsy from 38 Cd-exposed and 17 nonexposed subjects to determine how levels of these elements in bone are affected by Cd exposure and whether they are associated with the bone lesions due to Cd exposure, osteomalacia, and osteoporosis. Cd in ribs was significantly higher in the Cd-exposed subjects than in nonexposed subjects. Zn tended to be higher, while Cu, Ca, P, and Mg tended to be lower in the ribs of Cd-exposed subjects, though these differences were not statistically significant. Zn, Ca, P, and Mg were highly correlated with each other in both Cd-exposed and nonexposed groups, but the associations of Ca, P, and Mg in the ribs with Zn concentrations differed in subjects and controls. Ca to Zn ratios were low in the Cd-exposed subjects, and the higher the grade of osteomalacia, the lower the Ca/Zn ratio. The decrease in Ca/Zn ratio was significantly correlated with increases in Cd. Cu showed a significant positive correlation with Cd and significant inverse correlation with Ca, P, and Mg in the Cd-exposed group. Cu and its relation to other elements did not show any association with osteomalacia. In conclusion, Ca/Zn ratio in bone was related to Cd exposure and the degree of osteomalacia in the Cd-exposed subjects. PMID- 9356194 TI - Effects of indoor environmental factors on respiratory health of children in a subtropical climate. AB - This study was conducted to determine whether indoor environmental factors affected respiratory symptoms in 4164 primary school children in Kaohsiung rural areas of Taiwan. Information on respiratory health symptoms and characteristics of the housing was obtained using a written questionnaire, completed by the parents of children. Multiple logistic regression analysis examined the relationship between respiratory health symptoms (cough, wheezing, bronchitis, asthma, and allergic rhinitis) and housing factors. Home dampness was significantly associated with all respiratory health symptoms. Incense burning and mosquito repellant burning showed effects on the reporting of coughing symptoms. No apparent associations were found with the other indoor factors included in this study or respiratory health symptoms. We conclude that dampness in the home has a pronounced effects on respiratory health symptoms and is a new public health issue in subtropical areas. PMID- 9356195 TI - A case-control study of lung cancer mortality in six Gila Basin, Arizona smelter towns. AB - To investigate factors related to lung cancer mortality in six Arizona copper smelter towns, we identified 185 lung cancer cases and two matched controls per case from decedent residents during 1979-1990. Detailed information on lifetime residential, occupational, and smoking history was obtained by structured telephone interviews with knowledgeable informants. Interviews were completed for 82% of 183 eligible cases and 88% of the targeted number (366) of controls. Estimated historical environmental exposures to smelter emissions, based on atmospheric diffusion modeling of measured SO2 concentrations, were linked with residential histories to derive individual profiles of residential exposure. Occupational histories were characterized by potential exposure to smelter emissions, asbestos, and ionizing radiation. Conditional logistic regression was used to compare study factors in cases and controls with adjustment for potential confounding factors: gender, Hispanic ethnicity, and smoking. In overall and gender-specific analyses, no statistically significant associations were observed between lung cancer risk and any of the measures of residential exposure to smelter emissions considered (town of residence at time of death, highest level of exposure, and duration or cumulative exposure above background levels), or any of the estimated occupational exposures (definite or potential asbestos, potential ionizing radiation, definite or potential smelter). Among male residents of some, but not all, towns, there was some evidence of a positive association between lung cancer risk and reported copper smelter-related employment (reported as definite), with the highest risk observed for Miami, Arizona. This study provided little evidence of a positive association between lung cancer mortality and residential exposure to smelter emissions. Specific factors associated with the apparent heterogeneity in lung cancer risk across study towns cannot be identified in this community-based study. PMID- 9356196 TI - Air pollution and doctors' house calls: results from the ERPURS system for monitoring the effects of air pollution on public health in Greater Paris, France, 1991-1995. Evaluation des Risques de la Pollution Urbaine pour la Sante. AB - This study examines short-term relationships between doctors' house calls and urban air pollution in Greater Paris for the period 1991-1995. Poisson regressions using nonparametric smoothing functions controlled for time trend, seasonal patterns, pollen counts, influenza epidemics, and weather. The relationship between asthma visits and air pollution was stronger for children. A relative risk (RRP95/P5) of 1.32 [95% confidence interval (CI) = 1.17-1.47)] was observed for an increase from the 5th to the 95th percentile (7-51 micrograms/m3) in daily concentrations of black smoke (BS). The risks for 24-hr sulfur dioxide and nitrogen dioxide levels were in the same range. Cardiovascular conditions, considered globally, showed weaker associations than angina pectoris/myocardial infarction, for which RRP95/P5 was 1.63 (95% CI = 1.10-2.41) in relation to ozone ambient levels. Eye conditions were exclusively related to ozone (RRP95/P5 = 1.17, 95% CI 1.02-1.33). Asthma visits and ozone showed an interaction with minimum temperature: an effect was observed only at 10 degrees C or higher. In two-pollutant models including BS with, successively, SO2, NO2, and O3, only BS and O3 effects remained stable. Along with mortality and hospital admissions, house call activity data, available on a regular basis, may be a sensitive indicator for monitoring health effects related to air pollution. PMID- 9356197 TI - Herbicide/pesticide effects on intestinal epithelial growth. AB - The purpose of the present study was to examine the effects of some common herbicides and pesticides on the growth of normal intestinal and colonic epithelial cells. Preconfluent cultures of normal rat intestinal cells (IEC-6 cell line) and normal human colonic epithelial cells were treated with 0.05-50 microM doses of atrazine, diazinon, and endosulfan. After 3 days of treatment, the change in cell proliferation was quantified by cell counting or the MTT growth assay. Both intestinal and colonic epithelial cell cultures had increases in cell growth when treated with as little as 1.0 microM atrazine, diazinon, or endosulfan. The observed changes in both cultured intestinal and colonic cell growth rates were not due to the influence of the vehicle control dimethyl sulfoxide (DMSO). That is, the treatment of the cell cultures with concentrations of DMSO as high as 0.5% for 3 days resulted in no change in cell growth compared with untreated control cultures. A consistent observation with all three of the compounds was that the highest doses (50 microM) had the least "proliferative potential" in stimulating either IEC-6 cell or human colonic epithelial cell growth. Within the concentration range used, none of the herbicides or pesticides caused a decrease in cell proliferation below that of the untreated control cultures. Overall, treatment of IEC-6 cell cultures with atrazine, diazinon, or endosulfan produced a biphasic growth response, whereas the same treatment in the human colonic epithelial cell cultures produced a more sustained level of growth over the same period. This culture system may provide the basis for an in vitro model to further study the cellular and molecular basis of the effects of herbicides and pesticides on intestinal epithelial proliferation. PMID- 9356198 TI - Toxicokinetics of indomethacin-induced intestinal permeability in the rat. AB - Numerous studies in humans have demonstrated increases in intestinal permeability resulting from the administration of non-steroidal anti-inflammatory drugs (NSAIDs). The increased permeability correlates well with ulceration. The time course of the changes in intestinal permeability, however, has not been studied, which makes comparative studies between different NSAIDs or different formulations of the same drug difficult. In the present study we have administered single doses of indomethacin to examine both the time course and pharmacokinetic/pharmacodynamic relationships of intestinal permeability in rats estimated by following the urinary excretion of [51Cr]-EDTA. The change in intestinal permeability was both time- and dose-dependent. Following both 10 mg kg-1 and 20 mg kg-1 oral doses of indomethacin, there was a rapid rise in intestinal permeability to a maximum level, after at least 12 h post-dose, which is longer than those previously observed for ibuprofen, ketoprofen, flurbiprofen and naproxen. The maximal effect lasted 12 and 36 h following 10 and 20 mg kg-1 doses, respectively. The side-effect-plasma concentration relationship demonstrated a counter-clockwise hysteresis. The relationship between the observed side-effect and the estimated deep effect compartment concentration was, on the other hand, linear. In comparative permeability studies of NSAIDs the time of administration, concentration and drug dependencies should be considered. PMID- 9356199 TI - Hypocholesterolemic effects of Chinese tea. AB - Chinese teas with different degrees of fermentation were examined for their effect on diet-induced hypercholesterolemia in rats. The teas tested were Chinese Green tea, Jasmine, Iron Buddha, Oolong and Pu erh. Hypercholesterolemia was induced by feeding rats with a cholesterol-enriched diet for 1 week. They were then treated with different tea extracts together with a cholesterol-enriched diet for another 8 weeks. Chinese Green tea and Jasmine tea, both with a minimum degree of fermentation, were found to have significant serum and liver cholesterol lowering effects. They also reduced the increase in liver weight due to lipid deposition. All tea treatments lowered the atherogenic index and increased the HDL-total cholesterol ratio, while HDL-cholesterol and triglyceride levels were not significantly affected. Analysis of catechin levels in tea extracts showed that the individual catechin component in Chinese Green tea and Jasmine tea were significantly higher than the others. (-)-Epicatechin gallate and (-)-epigallocatechin gallate in the tea extracts may account for their hypocholesterolemic effect. PMID- 9356201 TI - Experimental kallikrein gene therapy in hypertension, cardiovascular and renal diseases. AB - Hypertension is a multi-gene and multi-factorial disorder affecting about 25% of the population. Hypertensive subjects are more likely to develop other cardiovascular diseases such as peripheral vascular disease, coronary heart disease, congestive heart failure and cerebrovascular disease. To demonstrate potential therapeutic effects of somatic gene delivery in treating hypertension, we delivered human tissue kallikrein in the form of naked DNA or in an adenovirus vector into hypertensive rats. Naked DNA constructs were delivered into spontaneously hypertensive rats via intramuscular, intravenous, intraportal vein and intraperitoneal routes. A single injection of human kallikrein DNA construct caused a sustained reduction of blood pressure which began 1 week post-injection and continued for more than 6 weeks. The hypotensive effect caused by somatic gene delivery of human tissue kallikrein in hypertensive rats is reversed by aprotinin, a potent tissue kallikrein inhibitor. Both systemic and local delivery of the human tissue kallikrein gene in an adenovirus vector were found to be highly effective in producing a rapid and sustained reduction of blood pressure in hypertensive rat models such as spontaneously hypertensive rats; two kidney, one clip Goldblatt hypertensive rats; and Dahl salt-sensitive rats. The expression of human tissue kallikrein in rats was identified in the heart, kidney, aorta, lung and liver by reverse transcription-polymerase chain reaction followed by Southern blot analysis and by ELISA. Adenovirus-mediated kallikrein gene delivery also resulted in the attenuation of glomerular and tubular damage and reduction of the left ventricular mass and cardiomyocyte size in Dahl salt sensitive rats fed a high salt diet. The ability of kallikrein gene delivery to produce a wide spectrum of beneficial effects makes it an excellent candidate in treating salt-related hypertension as well as cardiovascular and renal diseases. These results suggest the feasibility of applying somatic gene therapy for treating hypertension and salt-related cardiovascular and renal disorders. PMID- 9356200 TI - Pharmacology of the kallikrein-kinin system. PMID- 9356202 TI - EFFECT OF EARLY BLOCKADE OF BRADYKININ B2-RECEPTORS ON THE BLOOD PRESSURE PHENOTYPE OF NORMOTENSIVE AND SPONTANEOUSLY HYPERTENSIVE RATS AB - We evaluated if chronic blockade of bradykinin B2-receptors by the long-acting antagonist Icatibant (D-Arg,'Hyp3,Thi5,D-Tic7,Oic8'-bradykinin) affects blood pressure of rats. Pairs of normotensive Wistar Kyoto rats or spontaneously hypertensive rats were mated and their offspring received Icatibant (25 nmol day 1 per kg body wt., s.c.) or vehicle from the 2nd day until the 7th week of life. Then, the administration of Icatibant or vehicle was continued by i.p. infusion using Alzet osmotic pumps. At 9 weeks of age, normotensive rats given Icatibant showed greater systolic blood pressure (135±1 vs 115±1 mmHg in vehicle-treated rats, P<0.01), while heart rate was similar. The group difference regarding blood pressure levels was confirmed by direct intra-arterial measurement. No difference was detected between vehicle- and Icatibant-treated spontaneously hypertensive rats regarding blood pressure increase with aging. In conclusion, chronic blockade of bradykinin receptors by Icatibant alters the adult cardiovascular phenotype of Wistar Kyoto rats, provided that the antagonist is given at the early phases of life. These results suggest that the B2-receptor is essential for the maintenance of cardiovascular homeostasis during development, whereas it does not exert a protective role against the progression of hypertension in a rat model of genetic hypertension.Copyright 1997 The Italian Pharmacological Society 1997The Italian Pharmacological Society PMID- 9356203 TI - Kinins and cardioprotection. AB - During the past decade it was recognised that kinins are potent endogenous vasoactive peptides with important cardioprotective actions which are of therapeutic relevance in the acute and chronic beneficial effects of ACE inhibitors. The effect of kinins is mediated through the release of autocoids from the endothelium, particularly nitric oxide (NO). In this review article the cardioprotective effects of kinins and their importance for the therapeutic effects of ACE inhibitors are highlighted. PMID- 9356204 TI - Bradykinin and cardioprotection: don't set your heart on it. PMID- 9356205 TI - Kallikreins and kinins in inflammatory-like events in the reproductive tract. AB - The normal reproductive events of proliferation of the endometrial lining of the uterus during the menstrual cycle and ovulation have been likened to inflammatory like events. The kallikrein-kinin system is involved in inflammatory processes in many tissues. In this review, we identify which components of the kallikrein kinin system--the enzyme, tissue kallikrein; the substrate, low molecular weight kininogen and the effector receptor for the generated bradykinin peptide, the B2 receptor--have been identified in the uterus and ovary and their known involvement in the function of these organs. All three components have been localized to the glandular epithelial cells of the human endometrium. Tissue kallikrein gene expression is elevated midcycle when estrogens levels are also rising. This is also a time of extensive endometrial proliferation and tissue remodelling in preparation for embryo implantation, an event which is likened to other inflammatory processes. Similarly, tissue kallikrein gene expression was elevated following the estrogen surge at proestrous in the rat uterus, suggesting tissue kallikrein gene expression may be regulated by estrogens. Tissue kallikrein enzyme activity and gene expression has been demonstrated in the rat ovary and shown to be variously altered at the time of ovulation. Bradykinin has also been implicated in the expulsion of the ovum at the time of ovulation. These findings show that various components of the kallikrein-kinin system are present in the uterus and ovary. Further studies are required to more fully delineate their role in reproductive function. PMID- 9356206 TI - Effects of ethane dimethanesulfonate on the structure of adult male rat adrenal cortex. AB - The influence of ethane dimethanesulfonate (EDS), a specific toxicant for Leydig cells, on the morphometric characteristic of adult male rat adrenal cortex was examined on day 15 after administration. As expected, the dose of 75 mg kg-1 of EDS produced drastic reduction in serum testosterone levels followed by a decrease in testis and seminal vesicle weight. However, a considerable drop (over 50%) in adrenal weight was also found. Stereological analysis of the adrenal cortex, performed under light microscope, revealed atrophy of all adrenocortical zones. The changes were most prominent in the inner zones. Thus, in the zona fasciculata the number of parenchymal cells was markedly decreased. In the zona reticularis a layer consisting mostly of atrophic parenchymal cells was localised at the border between zona reticularis and zona fasciculata. The remaining small number of cortical cells in this zone displayed a notable hypertrophy. It is concluded that EDS at a dose that destroys Leydig cells has a strong deleterious effect on steroidogenic cells of adult male rat adrenal cortex. PMID- 9356207 TI - An explanation for failure to predict cyclosporine area under the curve using a limited sampling strategy: a beginner's second note. AB - Recently, limited sampling strategies have been developed to predict the area under a blood drug concentration-time curve (AUC). Multiple stepwise linear regression is the method used to develop limited sampling strategies. Several limited sampling strategies are used to predict cyclosporine AUC. However, recent findings demonstrated that, of all the developed limited sampling strategies, none can predict cyclosporine AUC correctly. Although several explanations have been given for this, the possible mathematical reason has not yet been fully explored. In the present study, we demonstrated that: (i) a steady-state blood drug concentration can be decomposed into the linear and non-linear components using the Fourier series; (ii) the non-linear component of the steady-state blood drug concentration leads to an AUC with a non-linear component, which affects the validation of limited sampling strategies; (iii) the correlation coefficient in a limited sampling strategy can only account for the linear association of linear components between a steady-state blood drug concentration and the AUC; (iv) the average steady-state blood drug concentration is a linear component of the steady state blood drug concentration; and (v) when developing a limited sampling strategy, the non-linear component can be effectively ignored by: (a) choosing the sampling time around the middle point between the peak and the trough steady state blood drug concentrations; and (b) increasing the trough steady-state blood drug concentration. However, the latter is restricted by clinical considerations. PMID- 9356208 TI - Separation of DL-dopa by means of micellar electrokinetic capillary chromatography after derivatization with Marfey's reagent. AB - Micellar electrokinetic capillary chromatography was used to separate levodopa (L 3,4-dihydroxyphenyl-L-alanine), dextrodopa (D-3,4-dihydroxyphenyl-L-alanine) and DL-3-O-methyl-dopa, a metabolite of dopa, which were derivatised with the Marfey's reagent, in sodium borate, sodium dodecyl sulfate and acetonitile buffer system. Whereas DL-dopa derivatives could be separated, DL-3-O-methyl-dopa derivatives could not. PMID- 9356209 TI - Antibacterial activity of rifaximin reduces the levels of benzodiazepine-like compounds in patients with liver cirrhosis. AB - Benzodiazepine-like compounds are present in trace amounts in the blood of normal subjects and increase in liver cirrhotic patients with or without encephalopathy. Their increased presence may, however, represent an occasional precipitating factor of hepatic encephalopathy. The source of these compounds is still unknown, but they are constituents of our diet since benzodiazepine receptor ligands have been described in plants, vegetables and in animals. They may also be synthesized, at least in part, by intestinal bacterial flora. In this article we report that the level of these compounds in the blood decreased by 40% after therapy with rifaximin, which reduces the aerobic and anaerobic intestinal bacterial flora. This observation indicates that intestinal bacterial flora is involved in the production of these compounds and that repeated short-term medications with this non-absorbable antibiotic may be useful in reducing the levels of benzodiazepine-like compounds in patients with liver cirrhosis. PMID- 9356210 TI - Antinociceptive effects of the hydrophilic alpha 2-adrenoceptor agonist ST-91 in different test circumstances after intrathecal administration to Wistar rats. AB - The antinociceptive and motor effects of the hydrophilic alpha 2-adrenoceptor agonist ST-91 were studied after intrathecal administration to male Wistar rats in different heat-pain tests and different test settings. Intrathecal administration of ST-91 caused a dose-dependent increase in hind paw licking latency in the hot-plate test, while in contrast with morphine it had a much lower efficacy in the tail-flick test in freely moving conditions. Sprague-Dawley rats gave similar results to those for Wistar rats in this setting. However, when the tail-flick test was performed under chronic restraint conditions (after a 1-h restraining period), the compound caused a significant antinociception. No signs of motor impairment and no changes in electromyographic activity were detected after ST-91 administration. The results indicate a characteristic analgesic profile for ST-91. In the interpretation of ST-91 data, consideration should be paid both to test model differences and to test conditions. PMID- 9356211 TI - cDNA cloning and chromosomal mapping of a predicted coiled-coil proline-rich protein immunogenic in meningioma patients. AB - There is increasing evidence that tumor expressed genes induce immune responses in cancer patients. To identify meningioma expressed antigens, we established a meningioma expression library which was screened with autologous serum. Out of 20 positive cDNA clones eight share high sequence homologies as determined by sequence analysis. These eight clones can be grouped into three classes which differ in length and which are characterized by specific sequence variations. The longest open reading frame was found to be 2412 bp encoding an immunoreactive antigen termed meningioma expressed antigen 6 (MEA6). Using five sequence specific primer pairs, somatic hybrid panel mapping revealed locations of the three classes on several human chromosomes including chromosomes 2, 3, 6, 7, 9, 13 and 14. The mapping results were confirmed by fluorescence in situ hybridization. RT-PCR showed consistent expression of all classes in several meningiomas and additional tissues using the same set of primer pairs as for chromosomal mapping. The expression data were confirmed by northern blot analysis. For the predicted amino acid sequence BLASTX revealed a homology to a human C219-reactive peptide which was previously isolated by an antibody directed against p-glycoprotein. Sequence properties of the MEA protein include an acidic activation domain, a proline-rich region and two coiled-coil domains indicating protein binding and activation functions. PMID- 9356212 TI - Effects of allopurinol on striatal dopamine, ascorbate and uric acid during an acute morphine challenge: ex vivo and in vivo studies. AB - In the present study in vivo and ex vivo experiments were combined to evaluate the effects of allopurinol on the neurochemical changes induced by an acute morphine challenge (2 mg kg-1, s.c.). In samples from rat striatum, levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 3 methoxytyramine (3-MT), ascorbate (AA), dehydroascorbate (DHAA), hypoxanthine, xanthine and uric acid (UA) were measured. Brain microdialysis experiments were carried out in freely moving rats. Striatal dialysate levels were assayed for DA, DOPAC + HVA, AA and UA using liquid chromatography followed by electrochemical detection. Morphine administration increased the striatal levels of DA metabolites, UA and DHAA and the extracellular concentrations of DA, DOPAC + HVA, UA and AA. Allopurinol (50 mg kg-1 by gavage), an inhibitor of xanthine oxidase which catalyses oxidation of xanthine to UA, decreased basal UA and AA concentrations and the morphine-induced increase in DA metabolites and AA oxidation. Since oxidation of DA and xanthines generates reactive oxygen species (ROS) and AA and UA are the main cellular antioxidants, these findings suggest that: (a) single morphine administration increases DA and xanthine oxidative metabolism with a consequent increase in ROS production, which may account for changes in concentrations of extracellular AA and tissue DHAA; (b) allopurinol decreases morphine-induced DA and xanthine oxidation; (c) UA and AA may act in concert to regulate levels of ROS in the brain. PMID- 9356213 TI - Proto-oncogene activity in the testis of the lizard, Podarcis s. sicula, during the annual reproductive cycle. AB - Since proto-oncogenes play a central role in the regulation of cellular growth and differentiation, localization of MYC, FOS, and JUN proteins has been studied in the testis of the lizard, Podarcis s. sicula, during the annual reproductive cycle by immunocytochemistry using antisera against c-myc, c-fos, and c-jun products. MYC was localized in the nuclei of spermatogonia (SPG), I and II spermatocytes (SPC), and spermatids (SPT). Strong immunoreactivity was detected in Sertoli cells just prior to the onset of the early spring spermatogenic wave coinciding with the androgen peak. FOS protein was present in the nuclei of SPG and SPC. In SPG an exclusive nuclear localization was seen during the active spermatogenic period (February-March and September). A perinuclear localization was observed during other months. Immunoreactivity in Sertoli cells was also observed during the periods of active spermatogenesis. JUN protein was localized in the cytoplasm of SPG as well as in I and II SPC and was detected in the nuclei of I and II SPC during April and October when spermatogenic waves occur. These data suggest that proto-oncogene activities have regulatory roles in the spermatogenesis of the lizard. PMID- 9356214 TI - Sites of estrogen receptor and aromatase expression in the chicken embryo. AB - Estrogens have been implicated in sexual differentiation of both the gonads and the genitalia of birds. In chicken embryos, the gonads are steroidogenically active from an early age, and the aromatase gene, (cAROM), necessary for estrogen synthesis, is expressed only in females at the time of gonadal sex differentiation. However, no studies have directly demonstrated the distribution of estrogen receptor (cER) transcripts or proteins in the embryonic avian reproductive system. Whole-mount in situ hybridization and immunohistochemistry were used here to identify sites of estrogen receptor expression in the embryonic chicken urogenital system. Estrogen receptor mRNA was observed in both male and female gonads prior to morphological differentiation, at Stage 26 (4.5 days of incubation), and continued until after sexual dimorphism at Stage 32 (7.5 days). Transcripts of cER were also detected in the Mullerian ducts and developing external genitalia of both sexes. Estrogen receptor protein was analysed in the embryonic gonads by immunohistochemistry and found to be most abundant in the cortex of the left ovary, although it was also present in the medulla of both female gonads. No significant cER protein expression was detected in the male gonad by immunohistochemistry. In contrast, the aromatase gene was expressed in the gonads of female embryos from the onset of sexual dimorphism but was not detectable in male gonads at any stage examined. These findings suggest that estrogen involvement in both gonadogenesis and genital development in chickens is mediated by the estrogen receptor. PMID- 9356215 TI - Down-regulation of TSH subunit mRNA levels by thyroid hormones in the European eel. AB - The effects of 3,5,3'-triiodothyronine (T3) and thyroxine (T4) on alpha and thyroid-stimulating hormone (TSH) beta subunit mRNA pituitary levels were examined in a teleost, the European silver eel. Northern blot analysis showed that the number and length of mRNAs encoding TSH beta varied among individuals, a variability apparently not related to thyroidal status. When several bands were present, their intensities were summed for quantitative analysis. Increasing circulating thyroid hormones (THs) by implantation of T3 or T4 significantly decreased TSH beta mRNA levels. Depression of circulating THs by thiourea treatment increased alpha and TSH beta mRNA levels. In vitro studies showed that T3 and T4 decrease TSH beta mRNA levels in primary cultures of eel pituitary cells. In conclusion, in vivo and in vitro experiments indicate that T3 and T4 exert a negative feedback action on pituitary TSH beta mRNA level in the European eel and that this effect might be exerted, at least partly, through a direct action on the pituitary. PMID- 9356216 TI - Elephantfish proinsulin possesses a monobasic processing site. AB - Total pancreatic RNA from the holocephalan species Callorhyncus milii (elephantfish) was used to make cDNA as a template for the polymerase chain reaction. Three redundant primers based on the known amino acid sequence of elephantfish insulin were used to amplify a fragment of proinsulin comprising truncated B-chain, complete C-peptide, and complete A-chain. Whereas the C peptide/A-chain junction contained the expected dibasic cleavage site (-Lys-Arg ), the B-chain/C-peptide junction was found to contain only a single Arg, the first such site to be unequivocally associated with the proteolytic processing of a proinsulin to insulin. Examination of the flanking sequences around this site shows that a typical endocrine/neuroendocrine PC3 conversion enzyme should still be able to cleave, as the general requirements for precursor processing at a monobasic site are satisfied, notably a basic residue (Lys) at the -4 position. An acidic residue (in this case Asp) at the +1 position, which is seen in all known proinsulins, is maintained. The corresponding genomic DNA fragment of elephantfish proinsulin was also amplified by PCR, revealing a 402-bp intron at the conserved IVS-2 position within the C7 codon. PMID- 9356217 TI - Purification of gonadotropin II from a teleost fish, the hybrid striped bass, and development of a specific enzyme-linked immunosorbent assay. AB - A highly purified gonadotropin II (GtH II), referred to as striped bass GtH II (stbGtH II), and its alpha and beta subunits were prepared from pituitaries of sexually mature hybrid striped bass. Pituitary glycoproteins were extracted with ethanol and intact stbGtH II purified by gel-filtration chromatography on Sephadex G-100, ion-exchange chromatography (IEC) on DE-52, and fast-performance liquid chromatography (FPLC) on Superdex 75. The presence of GtHs during the purification procedure was monitored by characteristic elution on reversed-phase high-performance liquid chromatography (rpHPLC) and in vitro steroidogenic activity. The stbGtH II alpha and beta subunits were purified from the pituitary ethanol extract by gel-filtration, IEC, and rpHPLC, and their identities assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), rpHPLC, and N-terminal amino acid sequencing. Molecular weights of intact stbGtH II and its alpha and beta subunits, determined by SDS-PAGE, were 34.5, 14.8, and 20.4 kDa, respectively. The stbGtH II beta subunit was used to produce specific antibodies, and a competitive enzyme-linked immunosorbent assay was developed using intact stbGtH II for the standard curve. The sensitivity of the assay was 156 pg/ml (15.6 pg/well) and the intra- and interassay coefficients of variation (at 50% binding) were 7.7% (n = 16) and 8.7% (n = 10), respectively. Physiological validation of the assay was performed by measuring changes of plasma GtH II levels in mature striped bass females, after injection of GnRHa ([d Ala6,Pro9-NEt]-mGnRH, 100 microg/kg BW). A maximum surge of GtH II in plasma was observed at 12 hr postinjection (22.5 +/- 3. 01 ng/ml), whereas GtH II levels in control fish (around 4 ng/ml) remained unchanged. Displacement curves obtained with serial dilutions of plasma and pituitaries from a number of perciform species were parallel to the standard curve, indicating that this assay can be used for GtH II measurements in a variety of fish species. PMID- 9356218 TI - Plasma gonadotropin II, sex steroids, and thyroid hormones in wild striped bass (Morone saxatilis) during spermiation and final oocyte maturation. AB - The blood levels of gonadotropin II (GtH II), sex-steroid hormones, and thyroid hormones were determined in wild spermiating male striped bass (Morone saxatilis) in males and in females at various stages of final oocyte maturation (FOM), captured on their spawning grounds. The progression of spermiation was associated with increases in plasma GtH II and decreases in plasma testosterone (T), 11 ketotestosterone, and thyroxine (T4). Plasma triiodothyronine (T3) remained at high and relatively unchanged levels. Plasma levels of 17,20beta-dihydroxy-4 pregnen-3-one (17,20beta-P) and 17,20beta, 21-trihydroxy-4-pregnen-3-one (17,20beta,21-P), the proposed maturation-inducing steroids (MIS) in striped bass, were low and unchanged during the same period. It was concluded that low progestogen levels are adequate to induce spermiation in striped bass, and that higher levels may be associated with spawning behavior. In the females, based on the profiles of the studied hormones, FOM was separated into two phases. Early FOM, which included germinal vesicle (GV) migration and lipid-droplet coalescence, was associated with elevations in plasma GtH II, T, and estradiol 17beta. Late FOM, which included GV breakdown and yolk-globule coalescence, was associated with a further surge in plasma GtH II, increases in the levels of the two MIS, mainly 17, 20beta-P, and a drop in T4. Plasma T3 levels did not change during FOM. Examination of conjugated steroids demonstrated, in the males, a reduction in conjugated androgens at the peak of the spawning season and, in the females, a small increase in conjugated 17, 20beta-dihydroxylated and 5beta reduced,3alpha-hydroxylated steroids after spawning. This is the most comprehensive report, to date, on the endocrine regulation of gonadal maturation in wild striped bass, demonstrating that a two-stage process of FOM is regulated by different endocrine signals, providing further evidence for the involvement of 17,20beta-P as a MIS in the females, and indicating that both males and females are in an euthyroid state during the spawning season. PMID- 9356219 TI - Increased circulating levels of testosterone and corticosterone in southern toads, Bufo terrestris, exposed to coal combustion waste. AB - This study describes an interrenal stress response in adult toads, Bufo terrestris, after exposure to coal combustion waste (characterized by a variety of trace elements). In the first portion of this study, free-ranging male toads captured at the coal ash polluted site exhibited significantly higher circulating levels of corticosterone (B) in both June/July and August than conspecifics captured at uncontaminated sites. In addition, both calling and noncalling males from the polluted site had higher B levels than conspecifics engaged in the same behaviors at reference sites. Testosterone levels were elevated in toads from the polluted site, regardless of capture month or behavioral state, suggesting altered androgen production, utilization, and/or clearance. In the second portion of this study, male toads from reference sites were transplanted to enclosures at the polluted site or an uncontaminated site. Toads held at the polluted site exhibited significant increases in B after 10 days of exposure compared to toads held at the reference site. B levels remained significantly elevated in toads transplanted to the polluted site after 12 weeks. We hypothesize that high concentrations of various trace elements in the polluted site are responsible for these hormonal responses. PMID- 9356220 TI - Immunoreactive gonadotropin-releasing hormone (GnRH) is detected only in the form of chicken GnRH-II within the brain of the green anole, Anolis carolinensis. AB - The presence of multiple forms of gonadotropin-releasing hormone (GnRH) within a single brain is common among vertebrate species. In previous studies of reptiles, two forms of GnRH were isolated from the brain of alligators and the primary structure was determined to be that of chicken (c)GnRH-I and cGnRH-II. GnRH has also been detected by indirect methods in other reptiles including turtles, lizards, and snakes. We used a combination of high-performance liquid chromatography (HPLC) and radioimmunoassay to determine the number and molecular form(s) of GnRH in the brain of a lizard, Anolis carolinensis, that was reported to lack GnRH cells in the forebrain. Immunoreactivity was detected in the same HPLC elution position in which synthetic cGnRH-II elutes, but not in any other position. Detection was based on five antisera that among them detect the 12 known forms of GnRH; these antisera include ones that are specific to cGnRH-I and cGnRH-II. We conclude that the lizard A. carolinensis contains cGnRH-II, but not cGnRH-I or another known form of GnRH. These data, coupled with our earlier immunocytochemical study, suggest that the lizard studied here lacks cGnRH-I, the form that is found in the terminal nerve, olfactory bulb, and forebrain in nonsquamate reptiles and in birds. Our hypothesis is that the presence of both cGnRH-I and cGnRH-II in the brain is ancestral in the reptilian lineage and retained in the orders that include turtles (Chelonia) or alligators (Crocodilia). However, the pattern in the order Squamata varies: in A. carolinensis, only cGnRH-II is present in the brain and cGnRH-I is absent, whereas in the snake Thamnophilis sirtalis, cGnRH-I is retained and cGnRH-II is absent in the brain, as recently reported. This raises the question of how reproduction is controlled in reptiles that lack one form of GnRH. PMID- 9356221 TI - Inactivation of Dip-allatostatin 5 by membrane preparations from the cockroach Diploptera punctata. AB - Incubation of Dip-AST 5 (Asp-Arg-Leu-Tyr-Ser-Phe-Gly-Leu-NH2) with membrane preparations of midgut, hindgut, brain, or corpora allata (CA) results in its inactivation in terms of the inhibition of juvenile hormone biosynthesis. Dip-AST 5 is initially cleaved at Gly7-Leu8 to yield the N-terminal heptapeptide (Asp-Arg Leu-Tyr-Ser-Phe-Gly). At supraphysiological concentration, the half-life of Dip AST 5 varied from 24 min by membrane preparations of brain to approximately 53 min following incubation with midgut membrane preparations. At more physiological concentrations (nanomolar), Dip-AST 5 was still initially cleaved to yield the inactive N-terminal heptapeptide with a half-life ranging from 23 min with brain membrane preparations to 85 min with membrane preparations of midgut. The fact that Dip-AST 5 is rapidly degraded to an inactive product by membrane preparations or whole tissues (CA) indicates that Dip-AST 5 has a different metabolic fate in tissue preparations than in diluted hemolymph (Garside et al., 1997). These findings demonstrate that the degradation of allatostatins by tissue preparations of D. punctata may play an important role in the termination of their ability to inhibit juvenile hormone biosynthesis by the CA and/or to modulate muscle activity in the hindgut. PMID- 9356222 TI - Asynchronous male and female gonadal cycles and plasma steroid concentrations in a viviparous lizard, Niveoscincus ocellatus (Scincidae), from Tasmania. AB - The reproductive cycle in males of the skink, Niveoscincus ocellatus, is characterised by testicular development during summer, followed by mating in autumn. Plasma testosterone concentrations show a bimodal seasonal cycle, with the major peak (18.6 +/- 1.2 ng/ml) in late summer/autumn and a minor peak (7.4 +/- 1.0 ng/ml) at spring emergence. In contrast to the males, the females have a gonadal cycle in which mating is temporally dissociated from peak development of the gonads: ovulation occurs in spring and the young are born in summer. Fresh mating marks on females in spring indicate that at least part of the population mates for a second time after spring emergence. In females, plasma estradiol concentrations are significantly elevated (956 +/- 214 pg/ml) through vitellogenesis and are highest (1241 +/- 175 pg/ml) during the preovulatory phase. Plasma progesterone concentrations rise during gestation to 6.5 +/- 1.5 ng/ml, but fall in the final stage of gestation to 1.6 +/- 0.2 ng/ml. There is minimal atresia of vitellogenic follicles, suggesting that clutch size is determined when the follicles are recruited for vitellogenesis. PMID- 9356223 TI - Immunocytochemical localization of vitamin D receptors in the shell gland of immature, laying, and molting hens. AB - It is accepted that vitamin D is involved in the control of egg calcification in hens. The goal of this study was to localize the vitamin D receptors (VDR) in hen shell gland and to determine whether their localization was dependent on reproductive function. Frozen sections of the shell gland of immature, laying, and molting hens were immunostained for VDR, and the VDR in these tissues were also examined by Western blot analysis. Both apical and basal cells of the mucosal epithelium as well as tubular gland cells showed a strong immunoreaction for VDR in the shell gland of laying hens. In the magnum and isthmus, the basal cells of the mucosal epithelium showed a moderately strong immunoreaction for VDR, whereas the immunoreactions in the apical cells of the mucosal epithelium and tubular gland cells were weak. In the shell gland of immature birds, both the mucosal epithelium and tubular gland cells showed a moderately strong VDR immunoreaction. In molting hens, the mucosal epithelial cells and tubular gland cells showed a strong VDR immunoreaction although the mucosal tissue was regressed. Western blot analysis indicated that the mucosal tissue of the shell gland of immature, laying, and molting hens contained two forms of immunoreactive VDR, which were approximately 58 and 60 kDa. Because VDR were richer in the shell gland than in other oviductal segments, these results suggest that in laying hens the shell gland tissues are one of the significant targets for vitamin D. It is likely that the amount of shell gland VDR increases during sexual maturation and immunoreactive VDR remain even during the molting phase. PMID- 9356224 TI - Fatty acids stimulate trehalose synthesis in trophocytes of the cockroach (Periplaneta americana) fat body. AB - Trophocytes from the disaggregated fat body of the cockroach (Periplaneta americana) respond to synthetic hypertrehalosemic hormone (HTH) by increasing the rate of trehalose synthesis. The cells give a similar response when incubated with stearic, oleic, linoleic, or arachidonic acid. A maximal increase in trehalose synthesis was obtained with 1-10 microM fatty acids. Synthesis of trehalose by the trophocytes was also increased by 1 microM prostaglandin F2alpha to nearly the same extent as that evoked by HTH. Furthermore, the data show that the trophocytes are capable of converting linoleic acid into arachidonic acid. This suggests that the cells may convert arachidonic acid, formed from the linoleic acid released by the action of HTH, to a prostaglandin which serves as an integral part of the hypertrehalosemic mechanism. PMID- 9356225 TI - Effect of melatonin on the response of the thyroid to thyrotropin stimulation in vitro. AB - Thyroidal-melatonin interactions are of particular importance to amphibian development since the thyroid controls the progress of metamorphosis while melatonin may coordinate its rate with prevailing environmental conditions. Melatonin antagonized thyroxine (T4) action at the tissue level and directly inhibited baseline T4 secretion in culture, so the present work sought to determine if it antagonized the response of the thyroid to thyroid stimulating hormone (TSH) as well. A preliminary experiment showed that, in Rana pipiens, the concentration of TSH (0.2 microg/ml) used in the culture of tadpole thyroids stimulated T4 secretion as much as frog pituitaries, but more than late premetamorphic tadpole pituitaries. There was no significant effect of 1 to 15 microg/ml melatonin in TSH-containing thyroid cultures of various Rana species of tadpoles and frogs in experiments with media collected once every 24 or 48 hr, although 15 microg/ml melatonin tended to depress T4 secretion. In a final experiment, a higher melatonin concentration was used as well as more frequent media collections. Thyroids from prometamorphic Rana catesbeiana tadpoles were cultured in L-15 media with periodic stimulation by 0.2 microg/ml TSH, or TSH and 10 or 100 microg/ml melatonin. Media were collected at the end of two 3-hr TSH pulses, and every 8 hr thereafter for the next 3 days. Melatonin was administered until the end of Day 2 while TSH was given only on Day 2 in addition to the original 3-hr pulses. The secretion of T4 was inhibited significantly by 10 microg/ml melatonin at only two of the early media collections. In contrast, 100 micro;g/ml melatonin significantly depressed T4 secretion in response to TSH at all but one interval and completely inhibited the thyroidal response to TSH reintroduced into the media on Day 2. The findings suggest that a high concentration of melatonin is inhibitory to the thyroidal response to TSH, but that lower concentrations do not significantly overcome the TSH stimulus. PMID- 9356226 TI - Unilateral ovariectomy influences hypothalamic monoamine asymmetries in a lizard (Anolis) that exhibits alternation of ovulation. AB - The lizard Anolis carolinensis alternates ovulation, and the resultant ovarian asymmetry correlates with alternating asymmetry of hypothalamic catecholamines. Unilateral and bilateral ovariectomies of cycling females were performed to determine if ovarian manipulations influence hypothalamic catecholamine asymmetries. During the middle of the ovarian cycle, we removed the larger ovary, i.e., the next one to ovulate an egg (N = 9), the smaller ovary with its corpus luteum (N = 8), or both ovaries (N = 5). A sham-operated control group was included (N = 9). The diameter of the largest ovarian follicle in each ovary was measured at the beginning and end of the experiment. After 12 days, the hemihypothalami from the sides of the initial smaller ovary (SO) and larger ovary (LO) were dissected and frozen for determination of monoamines and their metabolites using HPLC and electrochemical detection. Monoamine and metabolite concentrations at the end of the experiment in the original SO and LO sides of each hypothalamus were compared with an asymmetry ratio, or AR, of (SO side - LO side)/(SO side + LO side). No female ovulated during the experiment. Unilateral ovariectomy caused compensatory growth of the largest follicle in the remaining ovary. Removal of the SO or LO caused the AR of DOPAC to favor the brain side ipsilateral to that of the ovarian removal. Removal of the LO switched the NE AR from the SO to the LO side. Removal of the LO or SO caused the MHPG AR to favor the LO side. Ovariectomy of any kind caused 5-HT, which in the sham-operated animals favored the SO side, to become symmetric, and removal of the LO caused the 5-HIAA AR to favor the LO side. We conclude that the ovaries influence hypothalamic catecholamine asymmetries in Anolis via direct neural (as well as hormonal) pathways and that sensory input from the ovaries to the hypothalamus could be involved in control of ovarian alternation via both neural and hormonal efferent mechanisms. PMID- 9356227 TI - Purification, amino acid sequence, synthesis, and receptor selectivity of alligator gastrin. AB - Gastrin-like immunoreactive peptides were extracted from the gastric antrum of the American alligator (Alligator mississippiensis) and purified by fractionation using C18 Sep-Paks, Sephadex G-50, pH stable C8 reversed-phase HPLC, and C18 reversed-phase HPLC. Three major immunoreactive peaks were purified and found to correspond to 49, 45, and 34 residue peptides by microsequence analysis. The amino acid sequence of the largest peptide was DWLASLSQDQ KHLISKFLPH IYGELAN QEN YWQEDDALHD HDYPGWMDF-amide. The two smaller peptides corresponded to carboxyl terminal 45 and 34 residue fragments of the 49 residue peptide. The putative proteolysis of the 49 residue peptide to the two shorter peptides occurs at cleavage sites that are unusual in biosynthetic processing. Mass spectral analysis confirmed the molecular weights that were predicted from the amino acid sequences, thus revealing the absence of any post-translational modifications, such as sulfation. Although the three alligator gastrins resemble mammalian cholecystokinin in having a tyrosine residue in the seventh position from the carboxyl terminus, this tyrosine is apparently nonsulfated as in turtle gastrin. When tested by radioreceptor assay, a synthetic replicate of alligator gastrin-49 exhibited a gastrin-like pattern of biological activity on mammalian CCK-A and CCK-B receptors. Comparison of the amino acid sequences of known peptides revealed that alligator gastrin is most similar to turtle gastrin (76% identical), followed by frog gastrin (51% identical), chicken gastrin (49% identical), and human gastrin (12% identical). These similarities closely reflect vertebrate phylogeny and support the hypothesis that functionally distinct gastrins evolved from CCK in early tetrapods. However, gastrin evolved via different mechanisms in the mammalian lineage (mechanism unknown) versus the amphibian and reptilian/avian lineages, in which two different single nucleotide base changes can account for the separate evolution of amphibian gastrin and reptilian/avian gastrin. PMID- 9356228 TI - Characteristics of GnRH binding in the gonads and effects of lamprey GnRH-I and III on reproduction in the adult sea lamprey. AB - In the present study, both lamprey GnRH-I and -III stimulated steroidogenesis and induced ovulation in adult female sea lampreys during their final reproductive stage. One injection of lamprey GnRH-III at 0.1 or 0.2 microg/g lamprey stimulated plasma estradiol levels in lampreys held at each of three water temperatures, 13 degrees , 17 degrees , and 19 degrees , corresponding to increasing stages of maturation. Four successive injections, 3 to 4 days apart, of lamprey GnRH-III at 0.1 or 0.2 microg/g body weight induced ovulation in 100 or 88% of lampreys, respectively, compared to 21% in controls by Day 31. Lamprey GnRH-III also had a direct stimulatory effect on estradiol production in the sea lamprey gonads in vitro. Lamprey GnRH-III at 100 or 1000 ng/ml stimulated estradiol levels in media incubated with either lamprey ovaries or testes. In contrast to a previous finding in which lamprey GnRH-III was more potent than lamprey GnRH-I in inducing spermiation in adult male sea lampreys (Deragon and Sower, 1994), the results from the present study indicate that lamprey GnRH-I and -III are equally potent in inducing ovulation and stimulating steroidogenesis in female sea lampreys. In addition, GnRH binding sites have been demonstrated for the first time in both the testis and the ovary of the adult sea lamprey using an analog of mammalian GnRH ([D-Lys6] mammalian GnRH) as a labeled ligand. Scatchard analysis suggested the presence of a high affinity binding site in both the testis and the ovary. In summary, lamprey GnRH-III is biologically active in stimulating the pituitary-gonadal axis in adult female sea lampreys. This is the first report demonstrating the presence of a GnRH binding site in the gonads of an Agnathan. The evidence for a direct stimulatory effect of lamprey GnRH in the gonads, the presence of GnRH binding site, and the absence of GnRH in the plasma suggest that, like other vertebrates including rat, rabbit, teleost fish, and human, there may be a GnRH-like factor produced in the gonads of the lamprey and it may act as a paracrine/autocrine modulator of gonadal function. This study further strengthens the paracrine regulatory role of GnRH peptides in the gonads of vertebrates, which appear to be evolutionarily conserved. PMID- 9356229 TI - Role of platelet-activating factor in hepatocellular Ca2+ alterations during hemorrhagic shock. AB - A role of the potent proinflammatory Ca2+ agonist platelet-activating factor (PAF) on hepatocellular Ca2+ homeostasis and oxidant injury was investigated, since Ca2+ dysregulation has been demonstrated as a pivotal pathomechanism causing hepatic dysfunction during hemorrhagic or septic shock. In anesthetized male Sprague-Dawley rats, blood was withdrawn to a mean arterial blood pressure of 40 mm Hg for 60 min. Rats were resuscitated with 60% of shed blood and threefold the shed blood volume of lactated Ringers' (shock group). After 60 min of resuscitation, hepatocytes were isolated by portal collagenase perfusion. Hepatocellular Ca2+ uptake (Caup2+), initial rate of Ca2+ influx (Cain2+), and membrane Ca2+ flux (Caflux2+) were determined using 45Ca2+ incubation techniques. Hepatocyte oxidant injury was fluorometrically determined by thiobarbituric acid reactive substances. Caup2+ (3.37 +/- 0.15 vs. 2. 53 +/- 0.08 nmole Ca2+/mg protein), Cain2+ (0.42 +/- 0.1 vs. 0.27 +/- 0.02 nmole Ca2+/mg protein/min), and Caflux2+ (31.3 +/- 4.3 vs. 16.9 +/- 2.4 pmole Ca2+/mg/min) significantly increased in the untreated shock group compared to untreated sham-operated rats (P < 0.05). The specific PAF receptor antagonist BN52021 given 5 min before (5 mg/kg b.w.) and continuously during resuscitation (5 mg/kg/hr) significantly reduced Caup2+ in the shock group (2.73 +/- 0.2; P < 0.01) and prevented hepatocyte lipid peroxidation (shock: 91.9 +/- 1; shockBN52021: 66.7 +/- 2 nmole/mg wet weight; P < 0.01). These data suggest that platelet-activating factor plays a pivotal role in promoting hepatocyte Ca2+ overload during hemorrhagic shock by releasing Ca2+ agonistic mediators and inducing oxidative membrane alterations both of which are capable of enhancing cellular Ca2+ influx. PMID- 9356230 TI - Serum hyaluronic acid level reflects volume and ATP levels of the liver after extended hepatectomy with and without preoperative portal vein occlusion. AB - The purpose of this study was to examine the hypothesis that the serum hyaluronic acid (HA) level reflects the volume and adenosine triphosphate (ATP) levels of the liver after extended hepatectomy (ExHx) with and without preoperative portal vein occlusion (PVO). Rats were randomly divided into two groups, PVO-ExHx and sham-ExHx. At the first stage, they underwent PVO or sham surgery by an occlusion of the portal vein supplying the left lateral and median lobes or by similar manipulation but without PVO, respectively. Seven days after first-stage surgery, both groups received ExHx exceeding portal vein-occluded lobes, which was the excision of the left lateral, median, and right lateral lobes of the liver reported as a 90% hepatectomy model. On Days 0, 1, 2, and 3 after ExHx, the serum HA concentrations, liver weights, and hepatic ATP levels were determined. Liver volumes were restored to similar levels in both groups, but restoration was faster in the PVO-ExHx group because preoperative PVO allowed the excised liver less volume. Lower serum HA levels were significantly associated with significantly higher hepatic ATP levels and with a lower mortality rate in the PVO-ExHx group. The serum HA level correlated significantly (P < 0.001) with the liver weight and hepatic ATP level. The serum HA level serves as a proxy for clinically important parameters following major hepatic surgery. PMID- 9356231 TI - Loss and recovery of liver regeneration in rats with fulminant hepatic failure. AB - We earlier described a model of fulminant hepatic failure (FHF) in the rat where partial hepatectomy is combined with induction of right liver lobes necrosis. After this procedure, lack of regenerative response in the residual viable liver tissue (omental lobes) was associated with elevated plasma hepatocyte growth factor (HGF) and transforming growth factor beta (TGF-beta1) levels and delayed expression of HGF and c-met mRNA in the remnant liver. Here, we investigated whether syngeneic isolated hepatocytes transplanted in the spleen will prolong survival and facilitate liver regeneration in FHF rats. Inbred male Lewis rats were used. Group I rats (n = 46) received intrasplenic injection of 2 x 10(7) hepatocytes and 2 days later FHF was induced. Group II FHF rats (n = 46) received intrasplenic injection of saline. Rats undergoing partial hepatectomy of 68% (PH; n = 30) and a sham operation (SO; n = 30) served as controls. In 20 FHF rats (10 rats/group), survival time was determined. The remaining 72 FHF rats (36 rats/group) were used for physiologic studies (liver function and regeneration and plasma growth factor levels). In Group I rats survival was longer than that of Group II controls (73 +/- 22 hr vs. 33 +/- 9 hr; P < 0. 01). During the first 36 hr, Group I rats had lower blood ammonia, lactate, total bilirubin, PT, and PTT values, lower activity of liver enzymes, and higher monoethylglycinexylidide (MEGX) production than Group II rats. In Group I rats, livers increased in weight at a rate similar to that seen in PH controls and showed distinct mitotic and DNA synthetic activity (incorporation of bromodeoxyuridine and proliferation cell nuclear antigen expression). Plasma HGF and TGF-beta1 levels in these rats decreased and followed the pattern seen in PH rats; additionally, c-met expression in the remnant liver was accelerated. Hepatocyte transplantation prolonged survival in FHF rats and facilitated liver regeneration. Even though the remnant liver increased in weight four times reaching 30% of the original liver mass, the transplant-bearing rats expired due to inability of the regenerating liver to support the rat. PMID- 9356232 TI - Studies on polymorphonuclear leukocyte bactericidal function III: the role of extracellular matrix proteins. AB - We investigated the effect of the extracellular matrix proteins fibronectin (Fn) and laminin (Ln) on polymorphonuclear leukocytes (PMN) bactericidal activity. Adherence of PMN to increasing concentrations of Ln significantly increased the killing of Escherichia coli after 240 min of adherence, while fibronectin significantly increased PMN staphlacidal activity after 240 min of adherence. The addition of IL-1beta and IL-8 but not TNF-alpha increased PMN bactericidal activity against E. coli when PMN were adhered to Ln, while TNF-alpha and IL-8 increased PMN bactericidal activity against Staphylococcus aureus when PMN were adhered to Ln. TNF-alpha increased PMN killing of E. coli when PMN were adhered to Fn, while only IL-1beta increased the killing of S. aureus when PMN were adhered to FN. Anti-VLA-3 (alpha3/beta1) monoclonal antibodies (mAbs) inhibited the effect of Ln on PMN bactericidal activity, while anti-VLA-5 (alpha5/beta1) mAbs inhibited the effect of Fn on PMN bactericidal activity. Progressive cross linkage of these two receptors led to a dose-dependent reduction in PMN bactericidal activity for both pathogens when PMN were adhered to Ln or Fn, respectively. These results demonstrate that extracellular matrix proteins +/- exogenously added cytokines have the capacity to regulate PMN bactericidal activity. The signals sent by these matrix proteins to increase PMN bactericidal activity are transduced primarily via separate alpha subunits of the beta1 integrin complex. Stimulation of these receptors might lead to potential upregulation of PMN bactericidal activity which would be potentially advantageous in vivo at sites of infection. PMID- 9356233 TI - NF-kappaB activation and modulation in hepatic macrophages during cholestatic injury. AB - Cholestatic liver injury induces an inflammatory response that follows the activation of hepatic macrophages. Constitutive activation of the transcription factor, NF-kappaB, was found in these macrophages over the course of hepatic injury. Since NF-kappaB activation has been shown to have a key role in the inflammatory process, the modulatory effects of the antioxidant, alpha-tocopherol succinate, and the glucocorticoid, dexamethasone, on NF-kappaB activation were examined in this study. Male Sprague Dawley rats underwent 2-7 days of common bile duct division and ligation (CBDL) or sham laparotomy. Hepatic macrophages were isolated by collagenase Pronase perfusion and purified by centrifugal elutriation. Activation was determined by electrophoretic mobility shift assay and ELISA. We determined that NF-kappaB activation in injured hepatic macrophages could only be inhibited by dexamethasone. Dexamethasone-mediated inhibition of NF kappaB activation required the synthesis of a regulatory protein since cycloheximide-treated cells were resistant to its effects. Furthermore, dexamethasone-treated hepatic macrophages showed elevated steady-state levels of IkappaB-alpha mRNA, suggesting the role of IkappaB-alpha as a potential regulatory mediator. Consistent with constitutive transcriptional activation we showed constitutive secretion of TNF-alpha from injured hepatic macrophages which could be inhibited by dexamethasone. These data show for the first time, in a biologically significant model of hepatic injury, constitutive activation of the key inflammatory transcription factor NF-kappaB and cytokine TNF-alpha. These results support an approach focused on the NF-kappaB/IkappaB-alpha pathway as a critical target for therapeutic intervention during hepatic injury, and the consideration of possible steroid-based therapies. PMID- 9356234 TI - Regional tissue blood flow and pH in the brain during deep hypothermic retrograde brain perfusion. AB - Deep hypothermic retrograde brain perfusion is used to protect the brain during aortic arch operations. However, all experiments have failed to demonstrate retrograde blood flow in the brain tissue. We developed an experimental model of sagittal sinus and simultaneous superior vena cava perfusion. Brain tissue blood flow was mapped with colored microspheres during deep hypothermic retrograde brain perfusion in 9 dogs. Regional brain pH was mapped photometrically using neutral red as a pH-indicating dye after 90 min of retrograde brain perfusion in 28 dogs and after 60 min of circulatory arrest in 8 dogs. Cerebral surface blood flow was also measured during retrograde brain perfusion. They were analyzed as functions of driving pressure between sagittal sinus and aorta. Total brain blood flow (ml/min/100 g) was 1.4 +/- 1.3, 3.8 +/- 2.6, and 4.6 +/- 2.6 when the driving pressure was 15, 25, and 35 mmHg, respectively (P < 0.05, 15 mmHg vs 25 mmHg). Regional cerebral blood flow (ml/min/100 g) with a driving pressure of 25 mmHg was 12.1 +/- 9.4, 7.0 +/- 5.6, 4.4 +/- 2.8, and 2.2 +/- 1.4 in the frontal cortex, anterior, mid, and posterior cerebrum, respectively. Cerebral cortex pH was 6.86 +/- 0.23, 7.15 +/- 0.18, and 6.46 +/- 0.13 after 90 min of retrograde brain perfusion with driving pressure of less than 20 mmHg, after that of above 20 mmHg, and after 60 min of circulatory arrest, respectively. Brain tissue pH, blood flows measured with microspheres, and laser flowmetry were highest when driving pressure was between 25 and 35 mmHg. We conclude that retrograde brain perfusion may provide maximum brain protection with driving pressure of 25 to 35 mmHg. PMID- 9356235 TI - Effect of dexamethasone on NF-kB activation, tumor necrosis factor formation, and glucose dyshomeostasis in septic rats. AB - Glucocorticoids are potent anti-inflammatory and immunosuppressive therapeutic agents. The protective effect of dexamethasone (DEX) on hepatic phosphoenolpyruvate carboxykinase (PEPCK) transcript level, hepatic NF-kB (nuclear factor-kB) activation, and serum tumor necrosis factor alpha (TNF) formation was investigated in peritoneal sepsis induced by cecal incision in rats. For the control the rats were sham-operated with laparotomies only. Each group (N = 6) was pretreated with either normal saline (NS) or DEX before surgery (NS/Sham, NS/Sepsis, DEX/Sham, and DEX/Sepsis). At 3 hr post cecal incision, DEX treatment inhibited sepsis-induced hepatic NF-kB activation by 23%, suppressed circulating TNF by 50%, reduced serum glucose by 36%, reduced hepatic glycogen depletion by 76%, and attenuated PEPCK mRNA level. These findings suggested that DEX treatment was beneficial in attenuating glucose dyshomeostasis and significantly inhibited two sepsis-induced inflammatory mediators, NF-kB and TNF, in the early phase of peritoneal sepsis. However, in the late (6 hr) septic phase, DEX treatment inhibited serum TNF by 69%, but had no effect on NF-kB activation, glycogen depletion, and PEPCK mRNA level suggesting liver function failure injury. PMID- 9356236 TI - Glucose-induced intestinal hyperemia is mediated by nitric oxide. AB - Glucose-induced absorptive hyperemia of the intestine has been well demonstrated through microsphere blood flow experiments. We have previously demonstrated that glucose, when applied topically to rat ileal epithelium, restores microvascular vessel diameters and blood flow following Escherichia coli bacteremia or hemorrhage/resuscitation. However, the mechanisms of this hyperemia are not completely understood. We hypothesize that nitric oxide is a mediator of the microvascular response to glucose exposure on the rat intestinal epithelium. METHODS: Male Sprague-Dawley rats, 200-225 g, were monitored for hemodynamic stability with mean arterial blood pressure and heart rate. A 2-cm segment of the terminal ileum with intact neurovascular supply was exposed for intravital videomicroscopy. Intestinal arteriolar diameters (A1D, inflow; and A3D, premucosal arterioles) and microvascular blood flow (A1Q) were measured following topical application of isoosmotic glucose or saline, with or without l-NAME (LN, 100 mM), a competitive inhibitor of nitric oxide synthase. Statistical analysis was performed by ANOVA followed by Tukey-Kramer honestly significant difference test. RESULTS: All data are expressed as mean percentage changes from baseline +/ standard error of the mean. Hemodynamic variables did not change during the experimental procedure and there were no significant differences among group baselines. Addition of isotonic glucose to the bath solution caused a significant increase in A3D that persisted throughout the experiment (at 30 min, 19.2 +/- 4.2 vs -3.9 +/- 4.5, P < 0.05). This vasodilation was blocked by topical administration of LN (3.1 +/- 2.9, P < 0.05). A1D remained at baseline levels (saline and glucose) or constricted (LN) in all groups. Topical LN also attenuated A1Q in both the saline and glucose groups. CONCLUSIONS: These data demonstrate that glucose-induced intestinal hyperemia is primarily characterized by premucosal A3 arteriole dilation in this model and that nitric oxide is a mediator of glucose-induced intestinal hyperemia. These findings suggest that either (1) glucose directly causes endothelial nitric oxide production or (2) epithelial cells transduce a vasodilatory signal through vascular endothelial derived nitric oxide during postprandial intestinal hyperemia. PMID- 9356237 TI - Engineering tissue-specific expression of a recombinant adenovirus: selective transgene transcription in the pancreas using the amylase promoter. AB - Recombinant adenovirus accomplishes highly efficient gene transfer in vivo. Adenoviral vectors would be more attractive vehicles for gene therapy if transgene expression was inducible and restricted to the target tissue. In these studies, we hypothesized that selective transgene expression of a recombinant adenovirus could be accomplished by using a tissue-specific promoter of transcription. A replication-defective adenoviral vector was engineered to express the lacZ marker gene under control of the murine pancreatic amylase promoter. Expression of this vector occurred exclusively in the pancreas in neonatal and adult mice, while a similar vector with a constitutive promoter accomplished transgene expression in several organs. Within the adenoviral construct, the amylase promoter retained its ability to be induced by dexamethasone and insulin. This model will serve as a paradigm for selective and inducible adenoviral transgene expression. PMID- 9356238 TI - Macrophage colony-stimulating factor accelerates wound healing and upregulates TGF-beta1 mRNA levels through tissue macrophages. AB - Macrophage colony-stimulating factor (M-CSF) is produced by many cell types involved in wound repair, yet it acts specifically on monocytes and macrophages. The monocyte-derived cell is thought to be important in wound healing, but the importance of the role of tissue macrophages in wound healing has not been well defined. Dermal ulcers were created in normal and ischemic ears of young rabbits. Either rhM-CSF (17 microg/wound) or buffer was applied to each wound. Wounds were bisected and analyzed histologically at Days 7 and 10 postwounding. The amounts of epithelial growth and granulation tissue deposition were measured in all wounds. The level of increase of TGF-beta1 mRNA level in M-CSF-treated wounds was examined using competitive RT-PCR. M-CSF increased new granulation tissue formation by 37% (N = 21, P < 0.01) and 50% (P < 0.01) after single and multiple treatments, respectively, in nonischemic wounds. TGF-beta1 mRNA levels in rhM-CSF treated wounds increased 5.01-fold (N = 8) over vehicle-treated wounds under nonischemic conditions. In contrast, no effect could be detected in ischemic wounds treated with rhM-CSF, and these wounds only showed a 1.66-fold increase in TGF-beta1 mRNA levels when compared to ischemic wounds treated with vehicle alone. GAPDH, a housekeeping gene, showed no change. As mesenchymal cells lack receptors for M-CSF, the improved healing of wounds treated with topical rhM-CSF must reflect a generalized enhancement of activation and function of tissue macrophages, as demonstrated by upregulation of TGF-beta. The lack of effect under ischemic conditions suggests that either macrophage activity and/or response to M-CSF is adversely affected under those conditions; this may suggest the pathogenesis of impaired wound healing at the cellular level. PMID- 9356239 TI - Efficiency of pentoxifylline in donor pretreatment in rat liver transplantation. AB - Donor pretreatment is a new concept in organ preservation. Pentoxifylline (PTX) has been reported to suppress the activation of Kupffer cells and to decrease injury to the hepatic graft after rat liver transplantation. We evaluated the efficiency of PTX pretreatment on the donor against hepatic injury following cold ischemia (CI) or warm ischemia (WI) using the rat liver transplantation model. Dose dependency: every rat was injected intraperitoneally with PTX (30, 50, or 80 mg/kg) or saline. One hour later, the portal vein (PV) and the hepatic artery (HA) were clamped for 30 min. Transplantation: the donor rat was injected intraperitoneally with 50 mg/kg PTX or saline, 1 hr before laparotomy. Animals were divided into two groups. In the CI group, grafts were preserved for 12 hr in University of Wisconsin solution at 4 degrees C and transplanted. In the WI group, the PV and the HA in the donor were clamped for 30 min before donor surgery, and the grafts were transplanted. Serum levels of tumor necrosis factor alpha (TNF-alpha), glutathione S-transferase-alpha (GST-alpha), and aspartate transaminase (AST) were measured at 30 min, 3 hr, and 24 hr after reperfusion of the PV. Compared with those of a control group, the serum levels of TNF-alpha, GST-alpha, and AST in the PTX-pretreated groups were significantly lower after both CI and WI at 30 min and further suppressed in the WI group at 24 hr. These results indicate that PTX pretreatment on the donor is effective for suppression of hepatic injury after both CI and WI. PMID- 9356240 TI - The N tails of histones H3 and H4 adopt a highly structured conformation in the nucleosome. AB - The histone N tails correspond to conserved amino acid sequences that are peripherally located in the nucleosome and undergo a variety of post-synthetic modifications during cell cycle. These N tails have been recently recognized as directly interacting with transcription-related proteins. We show here, based on circular dichroic evidence, that the N tails of both tetrameric histones H3 and H4 are highly organized as DNA-bound polypeptide segments in the nucleosome core particle, with about half of their residues, taken together, being alpha-helical. In contrast, the N tails of both dimeric histones H2A and H2B are found essentially in a random-coil conformation. The implications of these findings on nucleosome structure and recognition are discussed. PMID- 9356241 TI - Differential DNA replication origin activities in human normal skin fibroblast and HeLa cell lines. AB - A modification of the extrusion method for the isolation of nascent DNA from mammalian cells and a PCR-based assay has been used in order to compare the in vivo activities of DNA replication origins in different cell lines. Conventional PCR was firstly applied to detect the chromosomal activities of several known (origins associated with c-myc, hsp70, beta-globin, immunoglobulin mu-chain enhancer) and putative DNA replication origins (autonomously replicating sequences obtained from enriched libraries of human origins of DNA replication from normal and transformed cells) in four human cell lines (HeLa, NSF, WI-38 and SK-MG-1). Then, in nascent DNA samples from normal skin fibroblast (NSF) and HeLa cells, abundance of DNA sequences in the regions of five of these origins was determined by competitive PCR. Our results suggest that autonomously replicating sequences NOA3, S14, S3 and F15 are associated with functional chromosomal origins of replication. Quantitative comparison of origin activities demonstrates that origins associated with c-myc and NOA3 are approximately twice as active in HeLa cells as in NSF cells. The described approach can facilitate the identification of origins which may be differentially active in normal cells and transformed cells or in different cell types. PMID- 9356242 TI - NMR study of the exchange rate between two stacked conformers of a model Holliday junction. AB - Our investigations into the folding behavior of a series of small model Holliday junctions in the presence of magnesium ions revealed a sequence (junction J9a) displaying a 71/29 population ratio between the two differently folded stacked X conformers in slow exchange on the NMR chemical shift time scale. For the first time we report the rates of interconversion between two stacked X-conformers and their lifetimes as measured by chemical exchange (EXSY) NMR spectroscopy: k1 5.6 (+/-0.5) s-1, k-1 2.3 (+/-0.2) s-1. The corresponding lifetimes (tau=1/k) are: tau1 430 ms, tau2 180 ms and the conformational transition barrier amounts to DeltaGdouble dagger294 68 kJ/mol (16.2 kcal/mol). It is argued that this free energy barrier reflects the maximum barrier that separates stacked X-conformers in junction 9a from the unfolded structure. PMID- 9356244 TI - Peach latent mosaic viroid is locked by a 2',5'-phosphodiester bond produced by in vitro self-ligation. AB - Although some viroid-like satellite RNAs possess self-cleavage and self-ligation activities, we show that the peach latent mosaic viroid (PLMVd) is unique among all known viroids since it also has such activities. These catalytic activities should have important roles in the rolling circle replication of PLMVd. According to this proposed mechanism, self-cleavage of the multimeric strands occurs via hammerhead structures producing monomers possessing 2',3'-cyclic phosphate and 5' hydroxyl termini. In the most stable predicted secondary structure for PLMVd these two extremities are juxtaposed, in order for self-ligation to occur. To establish the nature of the phosphodiester bond produced by self-ligation, we followed the classical procedure of complete enzymatic RNA hydrolysis coupled with thin layer chromatography fractionation. Using this procedure, we report that the self-ligation of PLMVd transcripts produces almost exclusively the 2',5' isomer (>96%). Primer extension assays also revealed that reverse transcriptase can read througth this 2', 5' linkage, suggesting that it does not prevent further replication of the viroid. Moreover, we have observed that this 2',5' linkage is resistant to the debranching activity contained in nuclear extracts, as well as being capable of preventing further viroid self-cleavage. Thus, if viroids do indeed self-ligate in vivo, the resulting 2', 5'-phosphodiester bond could contribute to the stability of these RNA species. Finally, an analysis of both the sequence and the structural requirements for hammerhead self-cleavage and self-ligation suggests that these two RNA processes may be interrelated. We hypothesize that the intramolecular self-ligation which produces circular conformers may contribute to the circularization step of the rolling circle replication, while the intermolecular non-enzymatic ligation is a potential mechanism for the sequence reassortment of viroids and viroid-like species. PMID- 9356243 TI - Promoter-specific activation of gene expression directed by bacteriophage selected zinc fingers. AB - It has been shown that sequence-specific DNA-binding domains containing zinc fingers can be selected from libraries displayed on filamentous bacteriophage. The affinity and specificity of these peptides are well characterised in vitro, but few data are available to demonstrate specific DNA binding and discrimination between closely related DNA sequences in vivo. Transient transactivation assays were performed in mammalian cells, using expression plasmids which produce different amounts of a model transcription factor containing a phage-selected zinc finger DNA-binding domain, and reporter plasmids which carry systematic variations of the promoter sequence. When the intracellular concentration of the transcription factor was appropriate, activation of gene expression was absolutely dependent on a promoter having the same DNA sequence as that originally used to select the zinc finger domain by phage display. However, excessive intracellular concentrations of the transcription factor resulted in some less-specific DNA binding, leading to gene activation from similar promoters containing a maximum of two base changes. Thus, provided delivery is carefully controlled, highly specific control of gene expression in vivo can be achieved using artificial transcription factors containing phage-selected zinc finger DNA binding domains. PMID- 9356245 TI - Selective phage infection mediated by epitope expression on F pilus. AB - Proteins and peptides can be displayed on bacterial and bacteriophage surfaces as fusions to bacterial integral membrane proteins or phage coat proteins. We now report on the expression of peptide antigens on the surface of F pili, elaborated by F+ strains of Escherichia coli. The peptides were expressed as fusions to F pilin, the building block of the F pilus that is encoded by the traA gene on the F plasmid. Filamentous bacteriophage infection of E. coli is normally mediated by phage binding to pilin at the F pili tip. Expression of 13 to 15 amino acid long peptides on the F pilus completely blocked infection by derivatives of wild-type infectious M13 phage. However, when a phage displaying a specific recombinant antibody fragment was allowed to interact with F pili displaying an antigenic peptide a bacterial infection could be demonstrated. This infection, mediated by the antibody-antigen interaction, resulted in bacterial cells containing plasmids encoding both the protein and the ligand. In a model library, where a scFv antibody against the human cytomegalovirus AD-2 epitope was selected we achieved an enrichment of 2500 of phage carrying the specific antibody, indicating an efficient selective infection. PMID- 9356246 TI - An in vivo and in vitro structure-function analysis of the Saccharomyces cerevisiae U3A snoRNP: protein-RNA contacts and base-pair interaction with the pre-ribosomal RNA. AB - The structure and accessibility of the S. cerevisiae U3A snoRNA was studied in semi-purified U3A snoRNPs using both chemical and enzymatic probes and in vivo using DMS as the probe. The results obtained show that S. cerevisiae U3A snoRNA is composed of a short 5' domain with two stem-loop structures containing the phylogenetically conserved boxes A' and A and a large cruciform 3' domain containing boxes B, C, C' and D. A precise identification of RNA-protein contacts is provided. Protection by proteins in the snoRNP and in vivo are nearly identical and were exclusively found in the 3' domain. There are two distinct protein anchoring sites: (i), box C' and its surrounding region, this site probably includes box D, (ii) the boxes B and C pair and the bases of stem-loop 2 and 4. Box C' is wrapped by the proteins. RNA-protein interactions are more loose at the level of boxes C and D and a box C and D interaction is preserved in the snoRNP. In accord with this location of the protein binding sites, an in vivo mutational analysis showed that box C' is important for U3A snoRNA accumulation, whereas mutations in the 5' domain have little effect on RNA stability. Our in vivo probing experiments strongly suggest that, in exponentially growing cells, most of the U3A snoRNA molecules are involved in the 10-bp interaction with the 5'-ETS region and in two of the interactions recently proposed with 18S rRNA sequences. Our experimental study leads to a slightly revised version of the model of interaction proposed by J. Hughes. Single-stranded segments linking the heterologous helices are highly sensitive to DMS in vivo and their functional importance was tested by a mutational analysis. PMID- 9356247 TI - Affinity and specificity of trp repressor-DNA interactions studied with fluorescent oligonucleotides. AB - Fluorescence-based solution methods have been used to study the binding of the trp repressor of Escherichia coli to a series of oligonucleotides bearing all or partial determinants for high affinity specific binding. The tryptophan, salt concentration and competitor DNA dependence of the binding affinities was examined for these targets. Binding to a fluorescein-labeled 20 base-pair hairpin structure oligonucleotide, which contains a palindromic repressor binding site (GAACTAGTTAACTAGTAC) and is known to bind repressor in a 1 : 1 dimer-DNA complex, resulted in a protein concentration-dependent, competable static quenching of fluorescence in presence of co-repressor, l-tryptophan. The affinity recovered from the fits of these intensity profiles at 100 mM KCl was on the order of 4x10(8) M-1. In absence of co-repressor an increase in intensity at high repressor concentration (>10(-7) M) was observed. The salt concentration dependence of the specific binding of the holo-repressor to this oligonucleotide was approximately half as large as what would be predicted by the number of phosphate contacts in the crystal structures of the complex. Repressor binding to the fluorescein-labeled hairpin 20mer was compared with binding to a rhodamine labeled 36 base-pair oligonucleotide bearing two inverted structural half-sites GNACT separated by an eight base-pair spacer containing none of the natural intervening sequence. The rather low affinity observed for the 36mer revealed that the intervening sequence in the natural operators contains energetic specificity determinants. Binding to a rhodamine-labeled oligonucleotide bearing a completely non-specific sequence was shown to occur over the same concentration range (>100 nM), regardless of tryptophan concentration, whereas binding to sequences bearing partial specificity ratio between 100 and 1000, depending upon the salt concentration. Even in absence of added KCl, the specificity ratio of trp repressor was greater than 100, implicating a significant free energy contribution from non-electrostatic interaction forces. PMID- 9356248 TI - Streptomycin binds to the decoding center of 16 S ribosomal RNA. AB - Streptomycin, an error-inducing aminoglycoside antibiotic, binds to a single site on the small ribosomal subunit of bacteria, but this site has not yet been defined precisely. Here, we demonstrate that streptomycin binds to E. coli 16 S rRNA in the absence of ribosomal proteins, and protects a set of bases in the decoding region against dimethyl sulfate attack. The binding studies were performed in a high ionic strength buffer containing 20 mM Mg2+. The pattern of protection in the decoding region was similar to that observed when streptomycin binds to the 30 S subunit. However, streptomycin also protects the 915 region of 16 S rRNA within the 30 S subunit, whereas it did not protect the 915 region of the naked 16 S rRNA. The interaction of streptomycin with 16 S rRNA was further defined by using two fragments that correspond to the 3' minor domain of 16 S rRNA and to the decoding analog, a portion of this domain encompassing the decoding center. In the presence of streptomycin, the pattern of protection against dimethyl sulfate attack for the two fragments was similar to that seen with the full-length 16 S rRNA. This indicates that the 3' minor domain as well as the decoding analog contain the recognition signals for the binding of streptomycin. However, streptomycin could not bind to the decoding analog in the absence of Mg2+. This contrasts with neomycin, another error-inducing aminoglycoside antibiotic, that binds to the decoding analog in the absence of Mg2+, but not at 20 mM Mg2+. Our results suggest that both neomycin and streptomycin interact with the decoding center, but recognize alternative conformations of this region. PMID- 9356249 TI - Probing RNA-protein interactions using pyrene-labeled oligodeoxynucleotides: Qbeta replicase efficiently binds small RNAs by recognizing pyrimidine residues. AB - Binding of small RNAs by the RNA-dependent RNA polymerase of coliphage Qbeta was studied utilizing a fluorometric assay. A DNA oligonucleotide probe of sequence 5'-d(TTTTTCC) was 5'-end-labeled with pyrene. In this construct, the proximal thymine residues efficiently quench the fluorophore emission in solution. Upon stoichiometric binding of one probe per polymerase molecule, the pyrene steady state fluorescence increases by two orders of magnitude, the fluorescence anisotropy increases, and a long fluorescence lifetime component of 140 ns appears. With addition of replicable RNA, steady-state fluorescence decreases in a concentration dependent manner and the long lifetime component is lost. This observation most likely reflects displacement of the pyrene-labeled probe from the proposed nucleic acid binding site II of Qbeta replicase. The effect was utilized to access binding affinities of different RNAs to this site in a reverse titration assay format. In 10 mM sodium phosphate (pH 7.0), 100 mM NaCl, at 16 degrees C, equilibrium dissociation constants for different template midi- and minivariant RNAs were calculated to be in the nanomolar range. In general, the minus and plus strands, concomitantly synthesized by Qbeta replicase during replication, exhibited discriminative affinities, while their hybrid bound less efficiently than either of the single strands. Different non-replicable tRNAs also bound to the polymerase with comparable dissociation constants. By titration with DNA homo-oligonucleotides it was shown that the probed site on Qbeta replicase does not require a 2' hydroxyl group for binding nucleic acids, but recognizes pyrimidine residues. Its interaction with thymine is lost in an A.T base-pair, while that with cytosine is retained after Watson-Crick base-pairing. These findings can explain the affinities of RNA-Qbeta replicase interactions reported here and in earlier investigations. The sensitivity of the described fluorometric assay allows detection of RNA amplification by Qbeta replicase in real-time. PMID- 9356250 TI - A conformational transition at the N terminus of the prion protein features in formation of the scrapie isoform. AB - The scrapie prion protein (PrPSc) is formed from the cellular isoform (PrPC) by a post-translational process that involves a profound conformational change. Linear epitopes for recombinant antibody Fab fragments (Fabs) on PrPC and on the protease-resistant core of PrPSc, designated PrP 27-30, were identified using ELISA and immunoprecipitation. An epitope region at the C terminus was accessible in both PrPC and PrP 27-30; in contrast, epitopes towards the N-terminal region (residues 90 to 120) were accessible in PrPC but largely cryptic in PrP 27-30. Denaturation of PrP 27-30 exposed the epitopes of the N-terminal domain. We argue from our findings that the major conformational change underlying PrPSc formation occurs within the N-terminal segment of PrP 27-30. PMID- 9356251 TI - Distinct regions of bacterial flagellar switch protein FliM interact with FliG, FliN and CheY. AB - The FliG, FliM, and FliN proteins of the bacterial flagellar motor are believed to interact with one another to form the switch complex, which in turn is thought to interact with one of the chemotaxis proteins, CheY. In particular, FliM appears to be an intermediary between CheY and FliG: the current model suggests that CheY, when phosphorylated (CheY-P), binds to FliM and produces a conformational change in FliM that is propagated to FliG. The result of these interactions is to induce clockwise rotation of the flagellar motors and tumbling of the cell. Various genetic and biochemical studies have provided evidence that the switch proteins associate with each other and that CheY-P binds to FliM. Here, we have used affinity blotting to obtain direct evidence of interaction between Salmonella typhimurium FliM and FliN, FliM and FliG, and FliM and CheY-P. We have also examined the ability of various FliM deletion and truncation mutant proteins to bind to FliN, FliG, and CheY-P. From these data, we conclude that distinct regions of the FliM protein bind to each of these other proteins. We propose a model in which the N-terminal region of FliM binds to CheY-P, the middle region of FliM binds to FliG, and the C-terminal region binds to FliN. PMID- 9356253 TI - Structural and functional effects of PA700 and modulator protein on proteasomes. AB - Control and targeting of the proteolytic activity of the major intracellular protease, the proteasome, is accomplished by various regulatory protein complexes that may form higher-order assemblies with the proteasome. An activator of proteolytic activity, PA700, has been shown to have an ATP-dependent stimulatory effect on the peptidase activities of the proteasome, and another protein factor, the modulator, further enhances the effect of PA700. Here we show that the addition of PA700 endows the proteasome with the ability to cleave ubiquitinated proteins, a property associated with the previously isolated 26 S form of the proteasome. The modulator further stimulates this specific activity, without having any such effect on the proteasome alone. Using electron microscopy, we show that addition of PA700 causes the appearance of protein "caps" at one or both ends of proteasomes, forming structures that are indistinguishable from 26 S proteasomes. Quantitation of the numbers of uncapped, singly capped and doubly capped complexes indicates cooperativity in the association of PA700 with the two ends of the proteasome. Addition of modulator protein makes no further structural modification that is detectable by electron microscopy, but does cause an increase in the number of capped complexes visible at subsaturating concentrations of PA700. Hence PA700 converts the proteasome both functionally and structurally to the 26 S form, and the modulator promotes this transformation, apparently without stable association with the resulting complex. PMID- 9356252 TI - Structural and functional characterization of homo-oligomeric complexes of alpha and beta chaperonin subunits from the hyperthermophilic archaeum Thermococcus strain KS-1. AB - To elucidate the function of group II chaperonin, the gene for the chaperonin from the hyperthermophilic archaeum Thermococcus strain KS-1 was cloned and sequenced. Two distinct genes coding for chaperonin subunits, designated alpha and beta, were obtained, and their deduced amino acid sequences are highly homologous to those of group II chaperonins from other sources. The alpha and beta subunits were individually expressed in Escherichia coli. Both of the recombinant subunits assemble to constitute the homo-oligomeric double-ring complexes, which are prone to form large aggregates. The alpha aggregate is dissociated into the typical chaperonin ring complex by incubation in buffer containing 15% (v/v) methanol, while the beta aggregate cannot be dissociated. At high temperature, both of the recombinant complexes have weak ATPase activities. They are able to arrest refolding of a chemically denatured thermophilic enzyme in the absence of ATP, and refolding is resumed when ATP is supplemented. These results suggest that homo-oligomeric complexes of the archaeal chaperonin have activity. PMID- 9356254 TI - Polymorphism of bacteriophage T7. AB - For viruses made of nucleic acid and protein, the structure of the protein outer shell has, in the past, been found to be uniquely determined by the viral genome. However, here, non-denaturing agarose gel electrophoresis of bacteriophage T7 reveals two states of the mature T7 capsid; the conditions of growth are found to alter the population by T7 of these two electrophoretically defined states. Both states have been previously observed for a genetically altered T7 and they are observed here for wild-type T7. The average electrical surface charge density of a bacteriophage particle (delta) determines its state; the delta of particles in both states is negative. For a given condition of growth, the population of these two states is influenced by the extent to which the major T7 outer shell protein, p10A, is accompanied by its minor readthrough variant, p10B. Comparison of the two electrophoretic states reveals the following. (1) No difference in radius is present in the outer shell (+/-2%). (2) As the pH of electrophoresis is either increased or decreased from neutrality, the state becomes more highly populated for which delta is greater in magnitude (state 1). By changing the pH, some T7 particles are made to change state. (3) Particles in state 1 adsorb less quickly to host cells than do the particles in the alternative state (state 2). This latter observation suggests the hypothesis that state 1 evolved to reduce the probability of re-initiating an infection when conditions are not favorable for growth. This hypothesis is supported by the observation that, as conditions of growth become apparently more unfavorable, progeny increasingly populate state 1. PMID- 9356255 TI - Structure and conformation of helical nucleic acids: analysis program (SCHNAaP). AB - We present a new versatile program, SCHNAaP, for the analysis of double-helical nucleic acid structures. The program uses mathematically rigorous and fully reversible procedures for calculating the structural parameters: the Cambridge University Engineering Department Helix computation Scheme (CEHS) is used to determine the local helical parameters and an analogous procedure is used to determine the global helical parameters. These parameters form a complete set that conforms to the "Cambridge Accord" on definitions and nomenclature of nucleic acid structure parameters. In addition to the two standard Watson-Crick base-pairs, the program handles mismatched base-pairs and chemically modified bases. An analysis of the sugar-phosphate backbone conformation is included. Standardized base-stacking diagrams of each dinucleotide step with reference to the mid-step triad are generated. Structures are classified as one of the four polymorphic families, A/B, Z, W or R, although W- and R-DNA (two types of hypothetical structure) have yet to be observed experimentally. PMID- 9356256 TI - Structure and conformation of helical nucleic acids: rebuilding program (SCHNArP). AB - We present a program, SCHNArP, for rebuilding double-helical nucleic acid structures from a set of helical parameters. The parameter sets are based on mathematically reversible schemes that allow direct comparison of data from experimental X-ray crystal structures analyzed using the analysis program, SCHNAaP (see accompanying paper), and structures built using the rebuilding program, SCHNArP. The program uses either local CEHS helical parameters or global helical parameters. A number of standard parameter sets from the literature are included that allow comparison of oligomer and polymer structures generated using different models for sequence-dependent DNA bending. Exact atomic models are provided for the bases. Schematic models that trace the path of the backbone and use rectangular blocks for the bases can be generated. PMID- 9356257 TI - Formation of a denatured dimer limits the thermal stability of Arc repressor. AB - The thermal stability of the Arc repressor dimer normally increases with concentration because protein folding and subunit association are thermodynamically coupled. At Arc concentrations above 100 microM, however, thermal denaturation remains reversible and cooperative but tm does not continue to increase. In this concentration regime, thermally denatured Arc shows significantly reduced secondary structure and no evidence of a tightly packed core, but light scattering and fluorescence polarization studies indicate that the protein is dimeric. Higher order denatured oligomers are not observed and the stability of the non-native dimer is reduced by Arc mutations, indicating that non-native dimerization involves specific interactions between Arc subunits. PMID- 9356260 TI - Common core structure of amyloid fibrils by synchrotron X-ray diffraction. AB - Tissue deposition of normally soluble proteins as insoluble amyloid fibrils is associated with serious diseases including the systemic amyloidoses, maturity onset diabetes, Alzheimer's disease and transmissible spongiform encephalopathy. Although the precursor proteins in different diseases do not share sequence homology or related native structure, the morphology and properties of all amyloid fibrils are remarkably similar. Using intense synchrotron sources we observed that six different ex vivo amyloid fibrils and two synthetic fibril preparations all gave similar high-resolution X-ray fibre diffraction patterns, consistent with a helical array of beta-sheets parallel to the fibre long axis, with the strands perpendicular to this axis. This confirms that amyloid fibrils comprise a structural superfamily and share a common protofilament substructure, irrespective of the nature of their precursor proteins. PMID- 9356261 TI - Solution structure of the spectrin repeat: a left-handed antiparallel triple helical coiled-coil. AB - Cytoskeletal proteins belonging to the spectrin family have an elongated structure composed of repetitive units. The three-dimensional solution structure of the 16th repeat from chicken brain alpha-spectrin (R16) has been determined by NMR spectroscopy and distance geometry-simulated annealing calculations. We used a total of 1035 distance restraints, which included 719 NOE-based values obtained by applying the ambiguous restraints for iterative assignment (ARIA) method. In addition, we performed a direct refinement against 1H-chemical shifts. The final ensemble of 20 structures shows an average RMSD of 1.52 A from the mean for the backbone atoms, excluding loops and N and C termini. R16 is made up of three antiparallel alpha-helices separated by two loops, and folds into a left-handed coiled-coil. The basic unit of spectrin is an antiparallel heterodimer composed of two homologous chains, beta and alpha. These assemble a tetramer via a mechanism that relies on the completion of a single repeat by association of the partial repeats located at the C terminus of the beta-chain (two helices) and at the N terminus of the alpha-chain (one helix). This tetramer is the assemblage able to cross-link actin filaments. Model building by homology of the "tetramerization" repeat from human erythrocyte spectrin illuminates the possible role of point mutations which cause hemolytic anemias. PMID- 9356262 TI - Catalysis of protein folding by parvulin. AB - Recently a new family of prolyl isomerases was discovered, which is unrelated with the cyclophilins or the FK-506 binding proteins. Parvulin, the smallest member of this new family, is a protein with only 92 residues, but parvulin-like domains occur in several large proteins that are apparently involved in protein folding or activation processes. We show here that, in addition to its activity in assays with proline-containing tetrapeptides, parvulin catalyzes the proline limited folding of a variant of ribonuclease T1 with a kcat/Km value of 30,000 M 1 s-1. This value is much smaller than the kcat/Km value of 1.1x10(7) M-1 s-1 determined for the parvulin-catalyzed prolyl isomerization in the tetrapeptide succinyl-Ala-Leu-Pro-Phe-4-nitroanilide. Parvulin itself unfolds and refolds reversibly in a simple two-state reaction with a Gibbs free energy of stabilization of 28 kJ/mol at 10 degrees C. Most of the unfolded parvulin molecules refold in a slow reaction that involves prolyl isomerization and is catalyzed by cyclophilin, another prolyl isomerase. Moreover, parvulin accelerates its own refolding in an autocatalytic fashion, and the rate of refolding increases tenfold when the concentration of parvulin is increased from 0.5 to 3.0 microM. Apparently, small single-domain prolyl isomerases catalyze prolyl isomerization much better in short peptides than in protein folding reactions, presumably because the prolyl bonds are less accessible in refolding protein chains. It is possible that the additional domains of the large prolyl isomerases improve the affinity for protein substrates. PMID- 9356263 TI - Dynamic frequency shifts of complexed ligands: An NMR study of D--1-13C,1-2H glucose complexed to the Escherichia coli periplasmic glucose/galactose receptor. AB - The 13C multiplet structure of D--1-13C,1-2H-glucose complexed to the Escherichia coli periplasmic glucose/galactose receptor has been studied as a function of temperature. Asymmetric multiplet patterns observed are shown to arise from dynamic frequency shifts. Multiplet asymmetry contributions resulting from shift anisotropy-dipolar cross correlations were found to be small, with optimal fits of the data corresponding to small, negative values of the correlation factor, chiCD-CSA. Additional broadening at higher temperatures most probably results from ligand exchange between free and complexed states. Effects of internal motion are also considered theoretically, and indicate that the order parameter for the bound glucose is >/=0.9. PMID- 9356265 TI - Quantum treatment of intermolecular multiple-quantum coherences with intramolecular J coupling in solution NMR. AB - A recently introduced density matrix picture for dipolar effects in solution NMR (1996, J. Chem. Phys. 105, 874) gave complete solutions for intermolecular multiple-quantum coherences for single-component samples without scalar couplings. This paper, for the first time, shows that this quantum picture can lead to explicit signal expressions for multicomponent samples of molecules with internal scalar couplings (here assumed to generate a first-order spectrum) and long-range dipolar couplings. Experimental observation of a triplet in the indirectly detected dimension for a heteronuclear CRAZED sequence (13CHCl3 sample, ZQ or 2Q coherences) gives clear evidence that the coupling is due to the intermolecular dipolar coupling. We also make comparisons with classical pictures which introduce the dipolar demagnetization field in multicomponent spin systems. PMID- 9356266 TI - Fast spiral magnetic resonance imaging with trapezoidal gradients. AB - A modified spiral imaging technique is presented, in which the conventional sinusoidal gradient waveforms are replaced by trapezoidal ones. In addition to allowing a reduced data acquisition time, the new waveforms circumvent specific hardware restrictions on the minimum scan repetition time. PMID- 9356268 TI - Electron spin-nuclear spin cross-correlation effects on multiplet splittings in paramagnetic proteins. AB - The effects of cross-correlation between Curie spin-nuclear dipole and nuclear dipole-nuclear dipole interactions on the linewidths and resonance frequencies of the individual lines of an AX multiplet in paramagnetic systems have been calculated. The implication of the relaxation-induced frequency shift of the lines (dynamic frequency shift) for the accurate measurement of residual dipolar couplings in field-oriented systems has been discussed. Our simulations indicate that these effects may play a role in the precise measurement of residual dipolar couplings in systems which belong to the small and intermediate tumbling regime, i.e., correlation times less than 5 ns. PMID- 9356270 TI - Off-resonance irradiation effect in steady-state NMR saturation transfer. AB - The off-resonance irradiation effect (spill-over effect), occurring in steady state NMR saturation-transfer experiments, is studied theoretically and experimentally for a two-spin system in chemical exchange, when a contralateral irradiation is applied to record the reference spectrum. The relevant parameter chosen for this study is the saturation-transfer ratio. It is defined as the ratio between the value of one exchanging magnetization, obtained when saturating the other, and that of the same magnetization measured when applying a contralateral irradiation. The theoretical study is carried out via a model based on the Bloch equations modified for chemical exchange and expressed in a doubly tilted single rotating frame. The saturation-transfer ratio is expressed as a function of the saturating RF field magnitude. It is shown that the RF field applied off-resonance during the acquisition of the reference spectrum does not correct the experimental saturation-transfer ratio for the spill-over effect. In fact, the saturation-transfer ratio increases with the magnitude of this field. This result is qualitatively explained by the consequence of the effective magnetic fields' relative orientations upon the amount of exchanged magnetization. The validity of the theoretical description is tested experimentally with a solution of N,N-dimethylacetamide in which chemical exchange arises from internal hindered rotation. An experimental protocol is proposed to detect spill-over and correct it when necessary. The way to describe spill-over theoretically when more than two spins interact by chemical exchange and/or dipolar coupling is also given. PMID- 9356271 TI - Dynamic contrast-enhanced imaging and analysis at high spatial resolution of MCF7 human breast tumors. AB - High resolution, dynamic GdDTPA-enhanced images of MCF7 human breast tumors in immunodeficient mice were analyzed at pixel resolution. The analysis, based on a physiological model, was performed by applying a nonlinear least-square algorithm using a color coded scale. The final output mapped at pixel resolution capillary permeability times surface area and fraction of extracellular volume, for each tumor slice. In addition, the output included assessment of the fit to the model by determining the proportion of variability (R2) for each pixel. The spatial variation in the R2 values served to identify regions where the predominant mechanism of enhancement was leakage from the intravascular volume to the extracellular volume (R2 close to 1). In regions with low R2 other mechanisms of enhancement appear to be dominating presumably diffusion within the extracellular space. As expected, in necrotic regions lacking microcapillaries and identified by analyzing T2-weighted images of the same tumors, the model failed to fit the dynamic contrast enhanced data. The heterogeneous distribution of the determined pathophysiological features demonstrates the importance of recording and analyzing breast tumor images at high spatial resolution. PMID- 9356275 TI - Origin of the correlation time dependence of coherence transfer distortions in rotating frame cross-relaxation spectra. AB - HOHAHA distortions in conventional (CW) ROESY experiments are known to be prominent when the frequency of the spin-lock field is near the midpoint between the resonance frequencies of a pair of coupled spins. That the disappearance of these distortions with offset from the midpoint is more rapid for large molecules than smaller ones is less widely known, and less well understood. We provide a quantitative explanation of the latter phenomenon using a combination of theory and numerical simulations. The cause of this effect can be found in the differential relaxation of the various magnetization modes which are involved in HOHAHA transfer. These modes experience enhanced relaxation far from a Hartmann Hahn match. This enhancement is larger for molecules which have long correlation times. PMID- 9356276 TI - Determination of orientational anisotropy in glassy solids by 2D dipolar spectra with sample flipping. AB - A method is proposed for the quantitative measurement of orientational anisotropy in glassy solids based on 2D dipolar NMR spectra with sample flipping (dipolar DECODER experiment). Purely dipolar spectra are obtained by chemical shift refocusing by a multiple pulse sequence. The experiment is applied to an investigation of a doubly 13C-labeled sample of bisphenol-A polycarbonate deformed in a channel-die apparatus. The orientational distribution function is determined by an expansion of the distribution in terms of spherical harmonics up to degree four. PMID- 9356277 TI - Two-dimensional ct-HC(C)H-COSY for resonance assignments of smaller 13C-labeled biomolecules. PMID- 9356278 TI - Chloroform mode of action: implications for cancer risk assessment. AB - Risk assessment methodology, particularly pertaining to potential human carcinogenic risks from exposures to environmental chemicals, is undergoing intense scrutiny from scientists, regulators, and legislators. The current practice of estimating human cancer risk is based almost exclusively on extrapolating the results of chronic, high-dose studies in rodents to estimate potential risk in humans. However, many scientists are questioning whether the logic used in this current risk assessment methodology is the best way to safeguard public health. A major tool of human cancer risk assessment is the linearized multistage (LMS) model. The LMS model has been identified as an aspect of risk assessment that could be improved. One way to facilitate this improvement is by developing a way to incorporate a carefully derived, more biologically relevant mechanism of action data on carcinogenesis. Recent data on chloroform indicate that the dose-response relationship for chloroform-induced tumors in rats and mice is nonlinear, based upon events secondary to cell necrosis and subsequent regeneration as the likely mode of action for the carcinogenic effects of chloroform. In light of these data, there is a sound scientific basis for removing some of the uncertainty that accompanies current cancer risk assessments of chloroform. The following points summarize the critical data: (1) a substantial body of data demonstrates a lack of direct in vivo or in vitro genotoxicity of chloroform; (2) chloroform induces liver and kidney tumors in long-term rodent cancer bioassays only at doses that induce frank toxicity at these target sites; (3) the chloroform doses required to produce tumors in susceptible species exceed the MTD, often by a considerable margin; (4) cytotoxicity and compensatory cell proliferation are associated with the chloroform doses required to induce liver or kidney tumors in susceptible rodent species; (5) there are no instances of chloroform-induced tumors that are not preceded by this pattern of dose-dependent toxic responses; (6) it is biologically plausible that cytolethality leads to chronically stimulated cell proliferation and related events such as inflammation and growth stimulation which act to initiate and promote the carcinogenic process; and (7) the consistently linked cellular events of cytolethality and subsequent cell proliferation, for which doses of no adverse effect have been clearly shown to exist, are one of the biological prerequisites for chloroform-induced tumors in animals. Based on these data, it is inappropriate to extrapolate cancer risk from high doses that produce necrosis and regenerative cell proliferation to low doses that do not with a model that presumes genotoxicity and a linear dose-response relationship. The weight of the scientific evidence concerning chloroform-induced tumors in rodents is consistent with and supports a cancer risk assessment methodology based on mode of action as the basis for establishing regulatory standards for this compound. PMID- 9356279 TI - A multiyear workplace-monitoring program for refractory ceramic fibers: findings and conclusions. AB - Results of a monitoring program carried out by members of the Refractory Ceramic Fibers Coalition as part of a Consent Agreement with the U.S. Environmental Protection Agency to measure workplace concentrations of refractory ceramic fiber (RCF) are presented. More than 700 personal monitoring samples were collected and analyzed annually from workers in RCF production and processing plants, as well as from those employed by customers/end users. The data indicate that (i) approximately 90% of time-weighted average (TWA) workplace concentrations are below the industry's recommended exposure guideline of 1 fiber per cubic centimeter TWA; (ii) workplace concentrations vary with functional job category; (iii) concentrations are approximately lognormally distributed; (iv) workplace concentrations are generally decreasing; (v) there are significant differences in workplace concentrations among plants operated by both RCF producers and customers; (vi) equations can be developed to interconvert data analyzed using different measurement techniques and counting rules; (vii) usage of respirators varies with the functional job category of the worker and the average fiber concentration; and (viii) workplace samples differ from those used in animal inhalation experiments in terms of the ratio of respirable particles to fibers. PMID- 9356280 TI - Subchronic toxicity (90-day) study with para-nonylphenol in rats. AB - As a component to the risk assessment process for para-nonylphenol (NP; CASRN 84852-15-3), a 90-day study was conducted in rats following U.S. EPA TSCA guidelines and Good Laboratory Practice regulations. NP was administered to four groups of rats at dietary concentrations of 0, 200, 650, or 2000 ppm which corresponded to approximate dietary intakes of 0, 15, 50, or 150 mg/kg/day, respectively. There were 25 rats/sex/group in the control and high-dose groups and 15 rats/sex/group in the low- and middose groups. Ten of the 25 rats/sex in the control and high-dose groups were designated as recovery animals and were maintained on control diets for 4 weeks after completion of the 90-day exposure period to assess the reversibility of any effects which might be observed. To evaluate for the possible weak estrogen-like activity that has been reported for NP in a number of screening assays, estrous cyclicity was monitored using vaginal cytology during Week 8 of the study, and sperm count, motility, and morphology were evaluated at termination. In-life effects from NP exposure were limited to small decreases in body weight and food consumption in the 2000-ppm dose group. Postmortem measurements at Week 14 indicated a dose-related kidney weight increase in males and a decrease in renal hyaline globules/droplets in males from the high-dose group. The kidney weights showed complete recovery following the 4 week postdosing recovery period. Due to the small magnitude of the changes (i.e., all weights were within or near laboratory historical control values) and the lack of correlating clinical or histopathological changes, the kidney weight alterations were not considered toxicologically significant. The biological significance of reduced hyaline in the kidneys of male rats from the high-dose group is uncertain. Renal tubular hyaline is associated with the rat-specific protein, alpha-2u-globulin, and, therefore, this finding was not considered toxicologically relevant to humans. No other effects attributable to NP were observed. No changes were observed for estrous cycling, sperm evaluations, or effects on endocrine organs. NP, therefore, did not manifest any estrogen-like activity as measured in these parameters at dietary concentrations as high as 2000 ppm, the maximum dose administered in this study. Based on the minor findings for the 2000-ppm dose group, the NOAEL (no-observed-adverse-effect level) for NP in this study is considered to be 650 ppm in the diet, corresponding to an approximate intake of 50 mg/kg/day. PMID- 9356281 TI - Health risk assessment of drinking water contaminants in Canada: the applicability of mixture risk assessment methods. AB - The objectives of this article are: (i) to review the current approaches of Health Canada to the risk assessment of drinking water contaminants, and (ii) to examine the applicability of mixture risk assessment methods to drinking water contaminants. Health Canada's current approaches to drinking water risk assessment, like those of many regulatory agencies, focus almost solely on the effects of individual chemicals. As such, no formal method is currently used for developing mixtures guidelines or for modifying guidelines of individual chemicals to account for the possibility of the occurrence of interactions (supraadditive or infraadditive). Recent interest in the risk assessment of mixtures, at least in part, stems from concerns over the potential health risks of mixtures of very commonly occurring compounds in Canadian drinking water supplies, namely the disinfection by-products. Before any mixtures methods can be considered for incorporation into Health Canada's current approaches to the risk assessment of drinking water contaminants, it is essential to consider the limitations and data requirements of the various mixture risk assessment methods (i.e., whole mixture approach, similar mixture approach, components-based approaches, interactions-based assessment). Among the existing mixture risk assessment methods, the components-based and interactions-based approaches could be applicable to drinking water contaminants. Specifically, among the components based approaches, dose-addition, response-addition, and the toxic equivalency factor approaches are the most applicable ones for drinking water contaminants. Until an interactions-based, mechanistic risk assessment approach (e.g., physiological model-based approach) becomes available for routine use, the components-based approaches remain the default methods for consideration. Progress in the development and validation of an interactions-based risk assessment methodology should facilitate a more realistic assessment of risk due to drinking water contaminants without increasing the levels of uncertainty in risk estimates above those associated with existing single-chemical methods. PMID- 9356282 TI - Multiple myeloma and benzene exposure in a multinational cohort of more than 250,000 petroleum workers. AB - Case reports have suggested an association between benzene exposure and multiple myeloma. Because petroleum workers are exposed to benzene or benzene-containing liquids, studies of these workers provide an opportunity for investigating the relationship between benzene and multiple myeloma. A large number of cohort studies of petroleum workers have been conducted. However, few of them have reported results of multiple myeloma separately. One reason is that multiple myeloma is usually grouped with other lymphopoietic cancers in the analysis. Another reason is that multiple myeloma is relatively rare, and few individual studies are large enough to provide reliable risk estimates. To determine the risk of multiple myeloma in petroleum (refinery, distribution, production, and pipeline) workers, we have identified 22 cohort mortality studies of petroleum workers in the United States, the United Kingdom, Canada, and Australia. Authors of these studies were contacted, and data on the number of observed deaths and age-specific person-years of observation were requested. Data from individual studies were combined in a pooled analysis (meta-analysis). In addition to the pooled analyses, results for individual cohorts, most of which have never been reported before, are also presented. The combined multinational cohort consisted of more than 250,000 petroleum workers, and the observation period covered an interval of 55 years from 1937 to 1991. A total of 205 deaths from multiple myeloma were observed, compared to 220.93 expected, a total derived from respective national mortality rates. The corresponding standardized mortality ratio (SMR) was 0.93 and the 95% confidence interval (95% CI) was 0.81-1.07. Additional analyses were performed by type of facility and industrial process. Stratum-specific SMRs (95% CIs) were 0.92 (0.77-1.09) for refinery workers and 0.93 (0.69-1.23) for distribution workers. When individual cohorts were stratified by length of observation, no pattern was detected. The pooled analysis indicates that petroleum workers are not at an increased risk of multiple myeloma as a result of their exposure to benzene, benzene-containing liquids, or other petroleum products in their work environment. This conclusion is supported by cohort studies of workers in other industries who were exposed to benzene as well as by population-based case-control studies of multiple myeloma and occupational exposures. PMID- 9356283 TI - Randomized, double-blind, placebo-controlled, consumer rechallenge test of Olean salted snacks. AB - Olestra is a zero-calorie fat substitute that is neither digested nor absorbed. A randomized, double-blind, placebo-controlled, within-subject, crossover rechallenge study was conducted to compare the occurrence of gastrointestinal symptoms after ingestion of chips made with Olean brand of olestra or conventional triglycerides in subjects who had previously experienced gastrointestinal symptoms they attributed to consuming Olean. A total of 57 male or female subjects received 2 oz of Olean potato chips or triglyceride potato chips at each of four weekly site visits. The occurrence of gastrointestinal effects after product consumption was noted in follow-up telephone interviews 3 to 5 days after each visit. There was no significant difference in the frequency of any gastrointestinal symptoms (abdominal cramping, diarrhea, loose stools) following consumption of Olean chips or triglyceride chips, and the severity of diarrhea, loose stools, and abdominal cramping was similar. We conclude that consumption of a 2-oz serving of Olean chips is no more likely to result in reports of gastrointestinal symptoms than consumption of triglyceride snacks as a part of the usual diet, even in individuals who have claimed intolerance to Olean. The data suggest that subjects who previously experienced symptoms that they attributed to consuming products made with Olean may have mistakenly attributed their symptoms to these products. PMID- 9356284 TI - Review and analysis of the effects of olestra, a dietary fat substitute, on gastrointestinal function and symptoms. AB - Olestra, a dietary fat substitute, was recently made available to consumers in savory snacks in three cities. Early reports of gastrointestinal complaints attributed to olestra attracted media coverage and fostered confusion among physicians and consumers about the nature of olestra and its effects on the digestive system. We reviewed all published studies of olestra's gastrointestinal effects and all relevant unpublished studies submitted to the Food and Drug Administration. Each study was analyzed by a group of expert gastroenterologists and epidemiologists. The symptoms reported with olestra ingestion are similar to those reported with ingestion of fiber and sorbitol, although the mechanisms involved in changing stool characteristics differ among these food additives. Olestra's effects on stool habit and characteristics are due to its presence in the stool. Large amounts are more likely to induce gastrointestinal symptoms than small amounts. There is no evidence that olestra induces pathological change in bowel function: there is no increased fluid or electrolyte nor is there altered gastrointestinal motility or microflora. Olestra and triglyceride ingestion resulted in a similar frequency of symptoms in normal adults and children and in people with chronic inflammatory bowel disease in remission. Olestra traverses the digestive tract intact to become a stool additive. Some subjects develop a change in bowel habit and stool characteristics due to the presence of more olestra in the stool. These changes resemble those associated with ingestion of sorbitol and fiber. PMID- 9356286 TI - Neonatal mouse assay for tumorigenicity: alternative to the chronic rodent bioassay. AB - The chronic rodent bioassay for tumors has been utilized systematically for 25 years to identify chemicals with carcinogenic potential in man. In general, those chemicals exhibiting tumorigenicity at multiple sites in both mice and rats have been regarded as possessing strong carcinogenic potential in humans. In comparison, the value of data collected for those test chemicals exhibiting more sporadic tumorigenicity results (e.g., single species/single sex or dose independent) has been questioned. As knowledge of the carcinogenic process has increased, several alternative test systems, usually faster and less expensive than the 2-year bioassay, have been suggested for identification of the strongly acting, transspecies carcinogens. The International Conference on Harmonization for Technical Requirements for the Registration of Pharmaceuticals for Human Use has proposed an international standard that allows for the use of one long-term rodent carcinogenicity study, plus one supplementary study to identify potential human pharmaceutical carcinogens. The neonatal mouse assay for tumorigenicity has been used since 1959; however, relative to other alternate tests, little has been written about this system. It is clear that this assay system successfully identifies transspecies carcinogens from numerous chemical classes, thus recommending itself as a strong candidate for a supplementary study to identify potential human carcinogens. In contrast, there are decidedly less data available from this assay in response to pharmaceuticals shown to exhibit weak and/or conflicting results in the 2-year bioassay, knowledge invaluable to the regulatory process. This paper reviews the historical development and our experience with the neonatal mouse assay and includes suggestions for a standardized protocol and strategies to document its response to "weak" and/or "nongenotoxic" carcinogens. PMID- 9356285 TI - Issues in setting health-based cleanup levels for arsenic in soil. AB - Health risk assessments often do not take into account the unique aspects of evaluating exposures to arsenic in soil. For example, risks from ingestion of arsenic in soil are often based on toxicity factors derived from studies of arsenic (soluble arsenate or arsenite) in drinking water. However, the toxicity of arsenic in drinking water cannot be directly extrapolated to toxicity of soil arsenic because of differences in chemical form, bioavailability, and excretion kinetics. Because of the differences between soil arsenic and water arsenic, we conclude that risks from arsenic in soil are lower than what would be calculated using default toxicity values for arsenic in drinking water. Site-specific risk assessments for arsenic in soil can be improved by characterizing the form of arsenic in soil, by conducting animal feeding or in vitro bioavailability studies using site soils, and by conducting studies to evaluate the relationship between urinary arsenic and soil arsenic levels. Such data could be used to more accurately measure the contribution that soil arsenic makes to total intake of arsenic. Available data suggest that arsenic usually makes a small contribution to this total. PMID- 9356287 TI - Hyaluronan-phospholipid interactions. AB - Hyaluronan-phospholipid interactions have been studied in vitro by negative staining and rotary shadowing electron microscopy. Hyaluronan (HA) molecules of different molecular weights (around 170,000; 740,000, and 1.9 x 10(6) Da) were added to phospholipid suspensions (DPPC or egg lecithin) that were in the form of either unilamellar particles or multilamellar vesicles. Suspensions were then gently stirred and incubated at different temperatures from 24 hr up to 7 days. After 24 hr, at temperatures just above the melting point of the phospholipid used, both unilamellar particles and multilamellar vesicles were already shown to change their organization in the presence of HA, giving rise to the formation of (1) huge perforated membrane-like structures lying on the substrate; (2) 12-nm thick "cylinders" (rollers) with a tendency to aggregate and to form sheets. These structures were seen only in the presence of high-molecular-weight HA, whereas low-molecular-weight HA (170 kDa) induced fragmentation of liposomes and formation of a few short rollers. These data show that phospholipids and HA interact and suggest they may also do so in vivo within the joint cavity, where both chemical species are present, giving rise to complexes which might exhibit peculiar lubricating and protective properties. It is also proposed that such interactions may not be as efficient in arthritic joints, where HA is degraded to low-molecular-weight fragments. PMID- 9356289 TI - The conformation of packaged bacteriophage T7 DNA: informative images of negatively stained T7. AB - Within the icosahedral protein outer shell of bacteriophage T7, a 40-kbp DNA genome occupies a cavity also occupied by a protein cylinder that projects into the DNA from the outer shell. However, neither the internal cylinder nor separately resolved DNA segments are revealed in the conventional negatively stained specimens of intact bacteriophage T7. In the present study, a procedure of negative staining is used that reveals both internal proteins and separately resolved segments of packaged DNA during electron microscopy of intact particles of a hybrid T7 bacteriophage; the hybrid is genetically T7, except for a tail fiber gene that has a segment from the T7-related bacteriophage, T3. The negatively stained packaged DNA segments of this hybrid bacteriophage are found to be wrapped around the axis of the internal cylinder. To obtain additional information about the conformation of packaged T7 DNA, electron microscopy is performed of negatively stained capsids that are incompletely filled with DNA (ipDNA-capsids); a procedure is described for improved isolation of ipDNA-capsids from lysates of hybrid bacteriophage T7-infected cells. The packaged DNA segments of ipDNA-capsids are found not to be wrapped around any axis. Images of ipDNA capsids are explained by the hypothesis that DNA does not achieve its wrapped condition until the capsid is more than 40% full of DNA. Wrapping via folding is, therefore, proposed to explain the images of DNA packaged in bacteriophage T7. PMID- 9356288 TI - Atomic force microscopy of arthropod gap junctions. AB - Atomic force microscopy has been used to characterize gap junctions isolated from the hepatopancreas of Nephrops norvegicus. The major polypeptide of these gap junctions is ductin, a highly conserved 16- to 18-kDa protein. The hydrated gap junctions, imaged in phosphate-buffered saline, appeared as membrane plaques with a thickness of 14 nm, consistent with their being a pair of apposing membranes. The upper membrane was removed by force dissection using an increased imaging force. The thickness of the lower membrane was 6 nm, giving a separation or gap between the two membranes of 2 nm. High-resolution images show fine details of the force-dissected extracellular surfaces, as previously reported for vertebrate and heart gap junctions. In addition high-resolution AFM images show for the first time detailed substructure on the cytoplasmic face of hydrated gap junctions of either vertebrate or invertebrate. The plaques had particles on their exposed and force-dissected faces. These particles were packed in a hexagonal lattice (a = b = 8.9 nm on both faces) and had a diameter of approximately 6.5 nm, with a central, pore-like depression. Fourier maps calculated from the AFM data suggested that each particle was composed of six subunits. These images show a marked similarity to the widely accepted structure of the connexon channel of vertebrate gap junctions. PMID- 9356290 TI - A method for establishing the handedness of biological macromolecules. AB - When biological macromolecules are imaged in the transmission electron microscope (TEM), their inherent handedness is lost because the three-dimensional (3D) structure is projected onto a two-dimensional (2D) plane, and identical 2D projections can be made from either 3D enantiomer. Nevertheless, tilt experiments in the TEM can be used to determine handedness. These experiments have been performed successfully on negatively stained specimens. More recently, the method was applied to unstained, frozen-hydrated specimens imaged by means of cryoelectron microscopy (cryoTEM) methods. Tilt experiments involve recording two micrographs of the same particles at different tilt angles, computing enantiomeric reconstructions from particle images in one micrograph, predicting orientations of corresponding particles in the second micrograph, and comparing model projections with particle images in the second micrograph. In principle, this procedure can be used to determine the handedness of any biological macromolecule imaged by cryoTEM, provided the enantiomeric reconstructions are distinguishable. PMID- 9356292 TI - Ultrastructure of intermediate filaments of nestin- and vimentin-immunoreactive astrocytes in organotypic slice cultures of hippocampus. AB - Glial cells in rat hippocampal slices cultured for 4 weeks were examined with immunocytochemical and cryoelectron microscopical methods. Astrocytes possessing long processes were similarly stained with antibodies against nestin, vimentin, and glial fibrillary acidic protein as seen by confocal microscopy. The three antibodies also labeled intermediate filaments in these astrocytes. In order to examine the fine structure of these intermediate filaments, slices were rapid frozen for freeze-substitution and freeze-etching. By freeze-substitution the processes of the astrocytes were packed with large hundles of intermediate filaments. In rapid-freeze deep-etched slices, these filaments were often interconnected with filamentous cross-bridges. These cross-bridges were rather uniform in size and shape (mean 2.9 nm thick and 14.8 nm long). These results suggest that the filament network with these cross-linkers is important for shaping the long processes of nestin- and vimentin-immunoreactive astrocytes in slice cultures. PMID- 9356291 TI - Centrosome injury in cells infected with human cytomegalovirus. AB - The organization of the mitotic apparatus was studied in human embryo lung fibroblasts (HEL) and Vero cells at 4 days postinfection with human cytomegalovirus (HCMV) strain AD 169. The bipolar spindle was detected by immunofluorescence in p72-positive mitotic cells exhibiting a regular or C metaphase-like chromosome configuration. Electron-microscopic study of C metaphase-like cells revealed alteration of the centrosome structure which is characterized by the following features: (1) breakdown of the diplosome, (2) separation of the fibrillar material from centrioles, and (3) disruption of the centriolar cylinder. The spindle pole in the aberrant mitotic cells consisted of one or several foci of microtubules converging on the fibrillar aggregates. There are not any signs of the nuclear envelope reconstruction found in mitotic cells with highly condensed scattered chromosomes. Unlike in HEL cells, viral particles were not detected in Vero cells. A question arises as to whether centrosome injury is an integral part of the events leading to cell death unrelated to the reproduction of HCMV. PMID- 9356294 TI - Crystallization and characterization of bovine liver glutamate dehydrogenase. AB - Bovine liver glutamate dehydrogenase has been crystallized as an abortive complex with glutamic acid, NADH, and an inhibitor, GTP. Crystals of this complex were grown using the sitting drop vapor diffusion method with PEG 8000 as the precipitant and diffract to better than 2.5 A resolution. The crystals belong to the space group P2(1) with an entire enzyme hexamer in the crystallographic asymmetric unit. Self-rotation and self-Patterson functions clearly define the orientation and position of this hexameric enzyme. PMID- 9356293 TI - Crystallization and preliminary X-ray analysis of the class 1 alpha 1,2 mannosidase from Saccharomyces cerevisiae. AB - The alpha 1,2-mannosidase from Saccharomyces cerevisiae catalyzes the conversion of Man9GlcNAc2 to Man8GlcNAc2 during the formation of N-linked oligosaccharides and is a member of the Class 1 alpha 1,2-mannosidases conserved from yeast to mammals. The enzyme is a type II membrane protein and a recombinant form of the alpha 1,2-mannosidase from S. cerevisiae, lacking the transmembrane domain, has been expressed in Pichia pastoris and crystallized using the hanging drop vapor diffusion technique. The crystals grow as flat plates, with unit cell dimensions a = 57.5 A, b = 84.1 A, c = 107.1 A, alpha = beta = gamma = 90 degrees. The crystals exhibit the symmetry of space group P2(1)2(1)2(1) and diffract to a minimum d-spacing of 3.5 A resolution. On the basis of density calculations one monomer is estimated to be present in the asymmetric unit (Vm = 2.08 A3 Da-1). This is the first report of the crystallization of any glycosidase involved in N glycan biosynthesis. PMID- 9356296 TI - Diffraction contrast imaging of extracellular matrix components using zero-loss filtering. AB - The beta-chitin microfibrils from the deep-sea hydrothermal vent worm Riftia pachyptila were studied in both mature and fresh tubes experimentally obtained. The methods used were electron diffraction and electron diffraction contrast images, in conjunction with an electron-energy filter, operated in the zero-loss mode. In both studied samples, microfibrils are organized in successive layers, inside which they are parallel. However, the fresh tube is less densely packed and diffraction data show that these microfibrils may be in a partially hydrated state. These results indicate that a later step occurs in the compaction of the tube material once it has been extruded. Comparison of filtered and unfiltered diffraction patterns shows that zero-loss filtering significantly improves both working conditions and quality of diffraction recordings. PMID- 9356297 TI - Evidence that the tandem Ig domains near the end of the muscle thick filament form an inelastic part of the I-band titin. AB - In vertebrate striated muscle the titin/connectin molecule spans half a sarcomere, and in the I-band forms and elastic filament connecting the Z-line with the end of the thick filaments. The only part of the elastic filament that has been described in intact rest length muscle is a short extension to the thick filament observed in freeze-fractured cardiac muscle which has similarities to the end-filament of negatively stained isolated thick filaments. We report here further observations made in sections of rabbit psoas muscle. In very thin longitudinal sections thin extensions to the thick filaments some 0.11 micron long and 5-6 nm in diameter are seen. Transverse sections show that each thick filament has such an extension. Nothing similar is seen further into the I-band or at the Z-line. The common features of this structure in both cardiac and skeletal muscle suggest that it corresponds to a common sequence in their titins. Such a sequence is to be found in the 22 tandem Ig domains near the A/I junction. Taken together with other information about the arrangement of domains in this part of the sarcomere, this leads to a calculated length for the end extension of 104 nm. The length of the extension does not vary with sarcomere length between 2.2 and 3.0 microns and therefore it corresponds to an inelastic region of I-band titin over the physiological range. Each extension probably comprises part of three to six titin molecules depending on the complement of titin in the thick filament, as previously suggested. A polymer formed from several strands of Ig domains would make for a relatively rigid structure which would resist folding or stretching when subjected to the small passive forces which pertain over the physiological range of sarcomere lengths. The relationship of the N2-line with the end-filament has also been studied. The N2-line position varies with sarcomere length in an elastic manner. Only at short sarcomere lengths does the end of the end-filament coincide with the position of the N2-line. Taking into account recent work on the elasticity of titin in the I-band we conclude that the N2-line corresponds to part of the elastic PEVK region of titin and not the region of titin sequence designated N2. PMID- 9356295 TI - Electron density projection map of the botulinum neurotoxin 900-kilodalton complex by electron crystallography. AB - The 900-kDa botulinum neurotoxin complex serotype A has been crystallized by the lipid-layer two-dimensional crystallization technique. Based on the binding characteristics of the hemagglutinating portion of the complex, a number of ganglioside/ lipid mixtures were tested but only lactosyl ceramide/1-palmityl-2 oleoyl-sn-glycero-3-phosphocholine was found to crystallize the complex. The optimum lipid mixture contained 75 mass % lactosyl ceramide and 25 mass % 1 palmityl-2-oleoyl-sn-glycero-3-phosphocholine. Using protein concentrations from 5 to 500 micrograms/ml and pH and 5 acetate buffer, we have obtained crystals that diffract to better than 15 A when prepared in negative stain. A projection map with a resolution of 30 A was calculated with unit cell dimensions of a = b = 157 A and P3 symmetry. The complex is triangular in shape with six distinct lobes observed. Additionally, six smaller structures protrude from the triangular core. PMID- 9356298 TI - Crystallization and preliminary X-ray analysis of human D-dopachrome tautomerase. AB - D-Dopachrome tautomerase catalyzes the conversion of D-dopachrome to 5,6 dihydroxyindole. This protein has amino acid sequence homology with that of macrophage migration inhibitory factor (MIF), suggesting a pathophysiological role of this protein in inflammatory and immunological events. We previously determined the tertiary structure of MIF and revealed the functional and evolutional relationships of this protein to isomerase. However, the reaction mechanism of both proteins associated with the inflammatory response, immune system, or tautomerase activities in vitro have not yet been clarified. The tertiary structure of D-dopachrome tautomerase would provide insight into the molecular function and the mechanism of these proteins. In this study, we crystallized human D-dopachrome tautomerase by a hanging-drop vapor diffusion method. The crystals belong to the trigonal space group P3, with unit cell dimensions a = b = 84.2 A and c = 41.0 A. They contain three (or two) monomers in the asymmetric unit, corresponding to a VM value of 2.21 (or 3.32) A3 Da-1. The best crystals diffract X-ray to 1.6 A resolution using a synchrotron radiation source. Crystallization of the selenomethionyl derivative of the protein for applying the multiwavelength anomalous diffraction method was also successful. PMID- 9356300 TI - Crystallization and preliminary X-ray crystallographic study of the ribosomal protein S7 from Bacillus stearothermophilus. AB - Overproduction and crystallization of Bacillus stearothermophilus ribosomal protein S7 (BstS7), a primary 16S rRNA binding protein and also a translational repressor protein, have been performed to analyze its three-dimensional structure by X-ray crystallography. Ribosomal protein BstS7 was expressed in the cytoplasmic fraction of the E. coli cells and purified to homogeneity. This recombinant BstS7 was used to produce crystals with P2(1) symmetry that diffracted to 2.5 A resolution which are suitable for high-resolution X-ray crystallographic analysis. PMID- 9356299 TI - Crystallization and preliminary crystallographic analysis of the sarcosine oxidase from Bacillus sp. NS-129. AB - The sarcosine oxidase from Bacillus sp. NS-129 has been crystallized by vapor diffusion using ammonium sulfate as precipitant. The tetragonal crystals have P4(1)2(1)2 (or P4(3)2(1)2) symmetry with a = b = 177.0 and c = 72.6 A. The crystals probably contain two sarcosine oxidase molecules per asymmetric unit and diffract to at least 2.7 A resolution. PMID- 9356301 TI - Binding of arsenicals to proteins in an in vitro methylation system. AB - The dynamics of interactions between rat liver cytosolic proteins and arsenicals were examined in an in vitro methylation system that contained cytosol, glutathione, S-adenosylmethionine, and 1 microM -73As-arsenite. After incubation at 37 degrees C for up to 90 min, low-molecular-weight components of the assay system (<10 kDa) were removed by ultrafiltration and cytosolic proteins were separated by size-exclusion chromatography on Sephacryl S-300 gel. Five 73As labeled protein peaks were found in chromatographic profiles. The estimated molecular masses of 73As-labeled proteins eluting in the three earliest peaks were as follows: Vo, >/=1000 kDa; A, 135 kDa; and B, 38 kDa. Peak C eluted immediately before the total volume (VT) of the chromatographic column; peak D eluted after the VT. 73As bound to proteins was released by CuCl treatment and speciated by thin-layer chromatography. Amounts and ratios of inorganic As, methyl As, and dimethyl As associated with cytosolic proteins depended upon the incubation interval. Inorganic As was present in all protein peaks. Methyl As was primarily associated with peaks A and C; dimethyl As was associated with peaks B and C. To examine the effect of valence on the binding of methylarsenicals to cytosolic proteins, trivalent or pentavalent 14C-labeled methyl As or dimethyl As was incubated in an in vitro system designed to minimize the enzymatically catalyzed production of methylated arsenicals. Proteins in peaks A, B, and C bound preferentially trivalent methyl and dimethyl As. Peak D bound either trivalent or pentavalent methyl and dimethyl As. Protein-bound inorganic and methyl As were substrates for the production of dimethyl As in an in vitro methylation system, suggesting a role for protein-bound arsenicals in the biomethylation of this metalloid. PMID- 9356304 TI - Induction of growth arrest and DNA damage-inducible genes Gadd45 and Gadd153 in primary rodent embryonic cells following exposure to methylmercury. AB - Methylmercury (MeHg) is recognized as a significant environmental hazard, particularly to the development of the nervous system. Studies on the mechanism of MeHg-induced toxicity reveal that inhibition of cell cycle progression may be one way by which MeHg interferes with normal development. In this study, we utilized primary rodent embryonic neuronal cell (CNS) and limb bud (LB) cultures to determine the mRNA expression level of two genes involved in cell cycle arrest, Gadd45 and Gadd153, both during cellular differentiation and in response to MeHg exposure. A differential expression pattern of Gadd45 and Gadd153 was observed during CNS and LB differentiation in culture. However, both CNS and LB cells responded to MeHg exposure with a concentration-dependent increase in Gadd45 and Gadd153 mRNA. Previous studies have shown that MeHg exposure (2 microm) of CNS cells for 24 hr causes a fourfold decrease in the number of cells passing through the cell cycle. The present study shows that at the same exposure concentration, a five- to eightfold increase in Gadd45 mRNA levels and a two- to fourfold increase of Gadd153 was observed. Induction of Gadd45 was also noted in adult female mice chronically exposed to 10 ppm MeHg, a dose that caused developmental toxicity in vivo. Based on the known involvement of the Gadd genes in cell cycle arrest, activation of these genes could be one mechanism by which MeHg interferes with the cell cycle in adult and developing organisms. PMID- 9356303 TI - A physiologically based pharmacokinetic model for trichloroethylene and its metabolites, chloral hydrate, trichloroacetate, dichloroacetate, trichloroethanol, and trichloroethanol glucuronide in B6C3F1 mice. AB - A six-compartment physiologically based pharmacokinetic (PBPK) model for the B6C3F1 mouse was developed for trichloroethylene (TCE) and was linked with five metabolite submodels consisting of four compartments each. The PBPK model for TCE and the metabolite submodels described oral uptake and metabolism of TCE to chloral hydrate (CH). CH was further metabolized to trichloroethanol (TCOH) and trichloroacetic acid (TCA). TCA was excreted in urine and, to a lesser degree, metabolized to dichloroacetic acid (DCA). DCA was further metabolized. The majority of TCOH was glucuronidated (TCOG) and excreted in the urine and feces. TCOH was also excreted in urine or converted back to CH. Partition coefficient (PC) values for TCE were determined by vial equilibrium, and PC values for nonvolatile metabolites were determined by centrifugation. The largest PC values for TCE were the fat/blood (36.4) and the blood/air (15.9) values. Tissue/blood PC values for the water-soluble metabolites were low, with all PC values under 2.0. Mice were given bolus oral doses of 300, 600, 1200, and 2000 mg/kg TCE dissolved in corn oil. At various time points, mice were sacrificed, and blood, liver, lung, fat, and urine were collected and assayed for TCE and metabolites. The 1200 mg/kg dose group was used to calibrate the PBPK model for TCE and its metabolites. Urinary excretion rate constant values were 0. 06/hr/kg for CH, 1.14/hr/kg for TCOH, 32.8/hr/kg for TCOG, and 1. 55/hr/kg for TCA. A fecal excretion rate constant value for TCOG was 4.61/hr/kg. For oral bolus dosing of TCE with 300, 600, and 2000 mg/kg, model predictions of TCE and several metabolites were in general agreement with observations. This PBPK model for TCE and metabolites is the most comprehensive PBPK model constructed for P450 mediated metabolism of TCE in the B6C3F1 mouse. PMID- 9356302 TI - Inhibition of 8-hydroxyguanine repair in testes after administration of cadmium chloride to GSH-depleted rats. AB - The main goal of this study is to investigate the mechanism of cadmium (Cd) induced carcinogenesis by reactive oxygen species. Rats were divided into four groups and were treated with (i) saline (control), (ii) cadmium chloride (CdCl2), (iii) l-buthionine-[S, R]-sulfoximine (BSO, an inhibitor of GSH biosynthesis), and (iv) CdCl2 and BSO, respectively. They were euthanized at 0, 24, 48, and 72 hr after these treatments, and the lungs and testes were analyzed. After treatment with both CdCl2 and BSO, the testicular 8-OH-Gua level increased (48 hr), its repair activity decreased (48 and 72 hr), the GSH content was markedly suppressed (48 and 72 hr), the superoxide dismutase activities slightly (48 and 72 hr) decreased, and the lipid peroxidation level increased (24 and 72 hr) in the testes as compared to the control levels. These results suggest that under GSH-depleted conditions, CdCl2 inhibits 8-OH-Gua repair activity in the rat testis and 8-OH-Gua accumulates in the DNA, which may pertain to testicular carcinogenesis. PMID- 9356305 TI - Toxicity of cordycepin in combination with the adenosine deaminase inhibitor 2' deoxycoformycin in beagle dogs. AB - For 3 consecutive days, the nucleoside cordycepin (3'-deoxyadenosine) was administered as 1-hr iv infusions (0, 1, 4, 8, 10, or 20 mg/kg/day) to dogs. These doses were given 1 hr after a bolus iv injection (0.25 mg/kg/day) of 2' deoxycoformycin (dCF), a potent inhibitor of adenosine deaminase. The hypothesis was that dCF would affect the toxicity of cordycepin. Plasma adenosine deaminase activity was strongly inhibited during the dose period and for 5 days following the final dose of dCF. Dogs given cordycepin alone showed no drug-related toxicities. In dogs given only dCF, drug-related toxicity to lymphoid tissue (lymphopenia and thymus lymphoid depletion), thrombocytopenia, and decreases in food consumption were observed. Cordycepin in combination with dCF produced symptoms associated with severe gastrointestinal toxicity (decreased body weights, emesis, diarrhea, decreased food consumption, and necrosis of the gastrointestinal tract) and bone marrow toxicity (lymphopenia, thrombocytopenia, and depletion of hematopoietic cells). The gastrointestinal tract and bone marrow were sites associated with dose-limiting toxicities. In surviving dogs, most of the effects were reversible by Day 30. The maximum tolerated dose of cordycepin administered in combination with dCF was 8 mg/kg/day (160 mg/m2/day) given daily for 3 days. PMID- 9356306 TI - Promotion of enzyme altered foci in female rat 2,3,3',4,4',5-hexachlorobiphenyl. AB - The tumor promoting activity of 2,3,3',4,4',5-hexachlorobiphenyl (PCB 156) was studied in an initiation/promotion bioassay in female Sprague-Dawley rats initiated with N-nitrosodiethylamine after partial hepatectomy. PCB 156 (50, 300, 1500, or 7500 microg/kg body weight/week) was administered by once-weekly subcutaneous injections for 20 weeks. Some high dose animals were left without treatment for an additional 20 weeks to study posttreatment effects. The volume fraction of the liver occupied by glutathione S-transferase P-positive foci was significantly increased to 2.9, 3.3, and 12% at 300, 1500, and 7500 microg/kg body weight/week, respectively, compared to 1.2% in the controls. The volume fraction was 43% in the high dose group 20 weeks after treatment was stopped, probably reflecting the slow body clearance of PCB 156 as indicated by the sustained liver and adipose tissue concentrations. Treatment with PCB 156 following initiation caused decreased body weight gain, thymic atrophy, liver enlargement, induction of hepatic cytochrome P450 1A1/2 (CYP1A1/2) and CYP2B1/2 activities, histopathological effects, and increased activities of aspartate aminotransferase and gamma-glutamyltransferase in plasma. These results show that PCB 156 can enhance the growth of altered foci in rat liver and probably act as a tumor promoter of hepatocarcinogenesis. Based on promotional activity a relative potency of PCB 156 to 2,3,7, 8-tetrachlorodibenzo-p-dioxin of 0.0001-0.001 is proposed. PMID- 9356308 TI - Mycotoxin-induced elevation of free sphingoid bases in precision-cut rat liver slices: specificity of the response and structure-activity relationships. AB - Fumonisin B1 (FB1) is the predominant member of a family of toxic metabolites produced by several species of Fusarium and is commonly found on corn. FB1 is a potent competitive inhibitor of ceramide synthase, which catalyzes the conversion of sphinganine and sphingosine to ceramide. The resultant accumulation of free sphingoid bases and the disruption of sphingolipid metabolism is believed to be the mechanism of toxicity of the fumonisins. The objectives of this study were to determine the relative potency of analogs of FB1 to inhibit ceramide synthase and to determine whether the inhibition is specific to mycotoxins with fumonisin-like structures. Fumonisins B1, B2, B3, B4, C4, and TA toxin (a structurally similar mycotoxin produced by the tomato pathogen, Alternaria alternata f. sp. lycopersici) were approximately equipotent inhibitors. Hydrolyzed fumonisins B1, B2, and B3, which lack the tricarballylic side chains, were only 30-40% as potent as the parent toxins. N-acetylated FB1 (FA1) did not block ceramide synthase, suggesting that FA1 is nontoxic. Inhibition of ceramide synthase by fumonisin analogs did not appear to be related to the lipophilicity of the compounds, as determined by computer estimation of log P values. The ability of relatively high (10 and 100 microm) doses of other mycotoxins that bear no structural similarity to fumonisins, including aflatoxin B1, cyclopiazonic acid, beauvericin, T-2 toxin, sterigmatocystin, luteoskyrin, verrucarin A, scirpentriol, and zearalenone, to block ceramide synthase was also determined. All of the toxins tested were negative in the bioassay with the exception of fumonisins, indicating that disruption of sphingolipid metabolism is a specific cytotoxic response. PMID- 9356307 TI - Higher frequency of aberrant crypt foci in rapid than slow acetylator inbred rats administered the colon carcinogen 3,2'-dimethyl-4-aminobiphenyl. AB - Humans and other mammals such as rats exhibit a genetic polymorphism in acetyltransferase (NAT2) capacity, yielding rapid and slow acetylator phenotypes. The rapid acetylator phenotype has been associated with increased incidence of human colorectal cancer in some, but not all, epidemiological studies. In order to investigate this possible association, a rapid (F-344) and slow (WKY) acetylator inbred rat model was utilized to investigate the role of the acetylator genotype (NAT2) in the formation of aberrant crypt foci (ACF) following administration of colon carcinogens. Age-matched (retired breeder) female rapid and slow acetylator inbred rats received two weekly injections (50 or 100 mg/kg, sc) of 3,2'-dimethyl-4-aminobiphenyl (DMABP) or a single 50 mg/kg, sc, injection of 1,2-dimethyl-hydrazine (DMH). The rats were euthanized at 10 weeks and ACF were evaluated in the cecum, ascending, transverse, and descending colon, and rectum. ACF were observed in the colon and rectum, but not the cecum of rapid and slow acetylator inbred rats administered DMABP or DMH. ACF were more concentrated in the descending colon. ACF frequencies were significantly higher in colons of rapid than slow acetylator inbred rats administered DMABP, a colon carcinogen which is activated via O-acetylation catalyzed by polymorphic acetyltransferase (NAT2). At 50 mg/kg, ACF frequency in the distal colon was 2.29 +/- 0.57 in rapid acetylators versus 0.38 +/- 0.18 in slow acetylators. At 100 mg/kg, ACF frequency was 4.11 +/- 1.06 in rapid versus 1.57 +/- 0.48 in slow acetylators. ACF frequency did not differ significantly between rapid and slow acetylator inbred rats administered DMH, a colon carcinogen which is not metabolized by polymorphic acetyltransferase. The two inbred rat strains did not differ in hepatic microsomal phenacetin deethylase activity, which is a marker for CYP1A2 activity important for the activation of aromatic amines. These results support the hypothesis that rapid acetylator (NAT2) genotype is a risk factor in aromatic amine-induced colon carcinogenesis. PMID- 9356309 TI - Ozone-induced stimulation of pulmonary sympathetic fibers: a protective mechanism against edema. AB - Tropospheric ozone exerts well-described toxic effects on the respiratory tract. Less documented, by contrast, is the ability of ozone to induce protective mechanisms against agents that are toxic to the lungs. In particular, interactions between ozone and the sympathetic nervous system have never been considered. Using a model of permeability edema in isolated perfused rabbit lungs, we report here that, immediately after exposure of rabbits to 0.4 ppm ozone for 4 hr, the pulmonary microvascular responses to acetylcholine and substance P are completely blocked. Several lines of evidence, including partial inhibition of the ozone-induced protective effect by several drugs (alpha2- and beta-adrenergic antagonists, neuropeptide Y antagonist, guanethidine), measured levels of released catecholamines in blood and urine and the in vitro response of isolated lungs exposed to 0.4 ppm ozone all seem to suggest that ozone can stimulate pulmonary adrenergic fibers and induce the local release of catecholamines and neuropeptide Y, this resulting in transient protection against pulmonary edema. We also showed that, 48 hr after the exposure, ozone increased the baseline microvascular permeability and the response to low concentrations of acetylcholine. PMID- 9356310 TI - A mathematical model of production, distribution, and metabolism of melatonin in mammalian systems. AB - Melatonin is a neuroendocrine hormone which is currently receiving considerable attention as a treatment for jet lag, a treatment for insomnia and, by some, a possible "magic bullet" for delaying the effects of aging and preventing cancer. Production of melatonin is focused primarily in the pineal gland with very wide daily shifts in production controlled by the day/night cycle. The potential for increased disease as a consequence of lower or higher than average production of this hormone has not been well studied, although potential environmental agents may modulate circulating levels (e.g., electric and magnetic fields). In this manuscript, a physiologically realistic mathematical model for the production, distribution, and metabolism of melatonin is developed as a precursor to a future study of the role of chemicals and environmental agents in altering this system. Values for key aspects of the system (e.g., diurnal rates of production of the hormone in the pineal gland) were obtained from the literature and the model was validated against data on circulating levels. The mathematical equations and model parameters are presented. PMID- 9356311 TI - Additive estrogenic activities of a binary mixture of 2',4',6'-trichloro- and 2',3',4',5'-tetrachloro-4-biphenylol. AB - The estrogenic activity of 2',4',6'-trichloro-4-biphenylol (HO-PCB3), 2',3',4',5' tetrachloro-4-biphenylol (HO-PCB4), and an equimolar mixture of both compounds (HO-PCB3/HO-PCB4) was investigated in the 21-day-old B6C3F1 mouse uterus, MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 cells, and in a yeast-based reporter gene assay. Treatment of the animals with 17beta-estradiol (E2) (0.02 microg/kg/day x3) resulted in increased uterine wet weight, peroxidase activity and progesterone receptor binding. Treatment with 18, 73, 183 or 366 micromol/kg (x3) doses of HO-PCB3, HO-PCB4, or HO-PCB3/HO-PCB4 (equimolar) caused a dose dependent increase in estrogenic activity; a maximal-induced response was not observed at any dose and the activity of the mixture was additive. Binding of E2, HO-PCB3, HO-PCB4, and HO-PCB3/HO-PCB4 to the mouse uterine estrogen receptor (ER) was determined in a competitive binding assay using [3H]E2 as the radioligand. The IC50 values were 1.1 x 10(-8), 3.4 x 10(-6), 9.9 x 10(-7), and 4.25 x 10(-6) m, respectively. HO-PCB3 and HO-PCB4 maximally induced MCF-7 cell proliferation, rat creatine kinase, and human complement C3 (C3-LUC) reporter gene activity at concentrations of 10(-5) to 10(-6) m, and these compounds were 10(3) to 10(4) less potent than E2. The HO-PCB3/HO-PCB4 mixture was active at the high concentration (10(-5) m) and was additive for these responses. HO-PCB3 and HO PCB4 also exhibited estrogenic activity in human HepG2 cells cotransfected with C3-LUC and an ER expression plasmid, and the estrogenic activity of the HO-PCB mixture was additive. Similar results were obtained in yeast transformed with the human ER and a double estrogen responsive element upstream of the beta-gal reporter gene. The effects of variable ER expression on the potential synergistic interactions of HO-PCB3/HO-PCB4 were investigated in HepG2 cells cotransfected with C3-LUC (405 ng/well) and variable amounts of ER expression plasmid (270, 27, 2.7, or 0.27 ng/well). The results show that as ER levels decreased, the magnitude of the induction response by E2, HO-PCB3, HO-PCB4, and HO-PCB3/HO-PCB4 also decreased. However, the activities of the HO-PCB mixture were additive at high and low levels of ER. Similar results were obtained in MDA-MB-231 cells cotransfected with C3-LUC and variable amounts of ER expression plasmid. The results of this study demonstrate that for several estrogen-responsive assays in the mouse uterus; MCF-7, HepG2, and MDA-MBA-231 human cancer cells; and a yeast based-reporter gene assay, both HO-PCB3 and HO-PCB4 exhibited estrogenic activity. The estrogenic activity of an equimolar mixture of these compounds was additive at high and low levels of ER expression. PMID- 9356312 TI - In vitro methylation of inorganic arsenic in mouse intestinal cecum. AB - The capacity of mouse intestinal cecal microflora to methylate inorganic arsenicals (iAs) was examined in vitro under conditions of restricted bacterial growth. Cecal contents incubated under anaerobic conditions at 37 degrees C for 21 hr methylated up to 40% of either 0.1 microM arsenite (iAsIII) or 0.1 microM arsenate (iAsV). Methylarsenic (MAs) was the predominant metabolite; however, about 3% of either substrate was converted to dimethylarsenic (DMAs). Over the first 6 hr, the rate of methylation was several times greater for iAsIII than for iAsV. There was a 3-hr delay in the production of methylated metabolites from iAsV, suggesting that reduction of iAsV to iAsIII before methylation could be rate limiting. Over the concentration range of 0.1 to 10 microM of iAsIII or iAsV, there was an approximately linear increase in the production of MAs and DMAs. There was evidence of saturation or inhibition of methylation at 100 microM of either substrate. Substrate concentration had little effect on MAs/DMAs ratio. Incubation of cecal contents at 0 degrees C abolished methylation of either arsenical. Under aerobic or anaerobic conditions, cecal tissue homogenates produced little MAs or DMAs from either arsenical. Addition of potential methyl group donors, L-methionine and methylcobalamin, into cecal contents significantly increased the rate of methylation, especially for iAsV. Addition of glutathione, but not L-cysteine, had a similar effect. Selenite, a recognized inhibitor of iAs methylation in mammalian tissues, inhibited methylation of either substrate by cecal contents. These data suggest that cecal microflora are a high capacity methylation system that might contribute significantly to methylation of iAs in intact animals. PMID- 9356313 TI - Long-term maintenance of drug-metabolizing enzyme activities in rat hepatocytes after cryopreservation. AB - Recent studies have demonstrated that freshly isolated adult hepatocytes from various species can be hypothermically preserved for a short period or cryopreserved for a prolonged period before seeding in primary culture. This study was designed to determine whether rat hepatocytes could be maintained functional for a prolonged period after either hypothermic preservation or cryopreservation. Cold storage was carried out in University of Wisconsin solution (UW) and freezing in Leibovitz medium added with 10% fetal calf serum and 16% dimethyl sulfoxide. Rat hepatocytes were then set up either in pure conventional culture or in coculture with rat liver epithelial cells. Various functions were measured over 4- and 15-day periods, i.e., albumin secretion rate, deethylation of ethoxyresorufin and phenacetin, dealkylation of pentoxyresorufin, glucuronidation and sulfoconjugation of paracetamol, and N-acetylation of procainamide. No major differences were observed between unfrozen, frozen, and UW preserved cells. While in pure culture all the functions tested were markedly decreased after 3 or 4 days, they remained high over the 15-day period in coculture, being either maintained or increased after 7-12 days compared to initial values. These results clearly demonstrate that when maintained under suitable culture conditions, rat hepatocytes can fully recover after hypothermic preservation or cryopreservation and therefore represent a suitable in vitro model system for pharmacotoxicological studies. PMID- 9356314 TI - 4-Amino-2,6-dichlorophenol nephrotoxicity in the Fischer 344 rat: protection by ascorbic acid, AT-125, and aminooxyacetic acid. AB - A halogenated derivative of 4-aminophenol, 4-amino-2, 6-dichlorophenol (ADCP), is a potent nephrotoxicant and a weak hepatotoxicant in Fischer 344 rats. Although the mechanism of ADCP nephrotoxicity is unknown, ADCP could undergo oxidation to a reactive intermediate, such as a 4-amino-2,6-dichlorophenoxy radical or 2,6 dichloro-1,4-benzoquinoneimine, which can generate additional free radicals and/or covalently bind to cellular proteins. The toxic process might also be mediated by glutathione (GSH) conjugates of ADCP, as suggested for the mechanism of 4-aminophenol nephrotoxicity. In this study, the effects of modulators of oxidation and GSH conjugation-related metabolism or transport on ADCP-induced nephrotoxicity were examined. In one set of experiments, male Fischer 344 rats (four/group) were intraperitoneally (ip) administered ADCP (0.38 mmol/kg) only or coadministered an antioxidant, ascorbic acid (1.14 mmol/kg, ip) with ADCP. Administration of ascorbic acid markedly reduced both functional nephrotoxicity and morphological changes induced by ADCP. Administration of a gamma glutamyltransferase (GGT) inhibitor, l-(alphaS, 5S)-alpha-amino-3-chloro-4,5 dihydroxy-5-isoxazoleacetic acid (10 mg/kg, ip), or a cysteine conjugate beta lyase inhibitor, aminooxyacetic acid (0.5 mmol/kg, ip), 1 hr before ADCP (0.38 mmol/kg) challenge partially protected rats against ADCP nephrotoxicity. In contrast, administration of an organic anion transport inhibitor, probenecid (140 mg/kg, ip), 30 min before ADCP had little effect on ADCP nephrotoxicity. The GSH depletor, buthionine sulfoximine (890 mg/kg, ip), was given 2 hr prior to ADCP and only minimal protection was noted. In addition, the nonprotein sulfhydryl (NPSH) contents in renal cortex and liver were determined at 2 hr following the administration of ADCP only or ascorbic acid/ADCP. Ascorbic acid afforded complete prevention of the depletion of NPSH in the kidney and liver caused by ADCP administration and also prevented the elevation of renal glutathione disulfide content induced by ADCP. The results indicate that oxidation of ADCP appears to be essential to ADCP nephrotoxicity and that GSH or GSH-derived conjugates of ADCP may be partly responsible for the nephrotoxic effects of ADCP via a GGT-mediated mechanism. PMID- 9356315 TI - NR8383 alveolar macrophage toxic growth arrest by hydrogen peroxide is associated with induction of growth-arrest and DNA damage-inducible genes GADD45 and GADD153. AB - Breathing air exposes humans and other mammals to various toxic agents including oxidative contaminants associated with fine particles of less than 2.5 micron which may be deposited in the deep lung and have been implicated in the increased morbidity and mortality correlated with air pollution. Oxidative damage from inhaled particles may include damage to DNA, thereby adversely affecting the immunosurveillance provided by alveolar macrophages. Using the rat alveolar macrophage cell line NR8383, we demonstrated that cell proliferation was inhibited by exogenous hydrogen peroxide, an oxidant naturally produced in cellular respiration and phagocytosis. Mercaptosuccinate, a specific inhibitor of the antioxidant enzyme glutathione peroxidase, also inhibited cell growth. Genes known to be coordinatively regulated in response to growth arrest and DNA damage, GADD45 and GADD153, were induced compared to the housekeeping gene beta-ACTIN by equitoxic doses of hydrogen peroxide and mercaptosuccinate. Hydrogen peroxide treatment of cells in which glutathione peroxidase was inhibited by mercaptosuccinate resulted in even greater induction of both GADD genes. This approach using the NR8383 alveolar macrophage cell line provides a model for studying genotoxicity at the mechanistic level at which stress-responsive genes involved in growth arrest and DNA-damage response are modulated. PMID- 9356316 TI - Characterization of antioxidant properties of natural killer-enhancing factor-B and induction of its expression by hydrogen peroxide. AB - Natural killer-enhancing factor B (NKEF-B) belongs to a highly conserved family of recently discovered antioxidants. The role of NKEF-B as an antioxidant was demonstrated by its protection of transfected cells to oxidative damage by hydrogen peroxide. To further characterize the antioxidant properties of NKEF-B, we compared the sensitivity of a human endothelial cell line ECV304 and its transfectant, B/1 that hyperexpresses NKEF-B, to various oxidants. In addition, we investigated the changes in the expression of NKEF-B mRNA upon oxidative stress. We found that B/1 was significantly more resistant than the control cells to the oxidative stresses caused by t-butyl hydroperoxide (t-BHP) and methyl mercury (MeHg). In contrast, there was no difference in the sensitivity of B/1 and the control cells to sulfhydryl reactive agents, diethyl maleate and diamide. B/1 was also as sensitive as the control cells to buthionine sulfoximine. The expression of NKEF-B mRNA was induced when the parental cell line ECV304 was treated with 2 mm HP. The induction reached a maximum level around 2 hr and decreased to the basal level around 4 hr. NKEF-A mRNA was not induced by HP. These results demonstrate antioxidant activities of NKEF-B toward prooxidants such as alkyl hydroperoxide and MeHg. Together with its antioxidant activity, the induction of NKEF-B by HP indicates that NKEF-B is an important oxidative stress protein providing protection against a variety of xenobiotic toxic agents. PMID- 9356319 TI - Gene-Culture Coevolution and Sex Ratios: II. Sex-Chromosomal Distorters and Cultural Preferences for Offspring Sex AB - Cultural preferences for the sex of offspring may produce behavior, such as female infanticide, sex-selective abortion and sex-selective parental investment, which alter the sex ratio in a population. Empirical evidence suggests that some genetic sex-ratio distorters are located on the sex chromosomes. Interactions between cultural preferences and sex-linked sex-ratio distorters are examined. Criteria for the spread of cultural preferences and sex-chromosomal distorter alleles are derived analytically, and the coevolution of preferences and distorters is examined through numerical iteration. Evolutionary equilibria and trajectories of gene-culture interactions involving sex-chromosomal distorter alleles may produce severely male- or female-biased primary sex ratios and adult sex ratios in populations. Adult sex ratios, primary sex ratios, allele frequencies and the prevalence of cultural preferences in the population are sensitive to initial conditions and cultural transmission parameters. During the coevolutionary process phenoallelic association is observed in many cases and is associated with unusual dynamics. Copyright 1997 Academic Press PMID- 9356317 TI - Mineralogical features associated with cytotoxic and proliferative effects of fibrous talc and asbestos on rodent tracheal epithelial and pleural mesothelial cells. AB - Inhalation of asbestos fibers causes cell damage and increases in cell proliferation in various cell types of the lung and pleura in vivo. By using a colony-forming efficiency (CFE) assay, the cytotoxicity and proliferative potential of three mineral samples containing various proportions of fibrous talc were compared to NIEHS samples of crocidolite and chrysotile asbestos in cell types giving rise to tracheobronchial carcinomas, i.e., hamster tracheal epithelial (HTE) cells, and mesotheliomas, i.e., rat pleural mesothelial (RPM) cells. Characterization of mineralogical composition, surface area, and size distributions as well as proportions of fibers in all mineral samples allowed examination of data by various dose parameters including equal weight concentrations, numbers of fibers >5 micron in length, and equivalent surface areas. Exposure to samples of asbestos caused increased numbers of colonies of HTE cells, an indication of proliferative potential, but fibrous talc did not. RPMs did not exhibit increased CFE in response to either asbestos or talc samples. Decreased numbers of colonies, an indication of cytotoxicity, were observed in both cell types and were more striking at lower weight concentrations of asbestos in comparison to talc samples. However, all samples of fibrous minerals produced comparable dose-response effects when dose was measured as numbers of fibers greater than 5 micron or surface area. The unique proliferative response of HTE cells to asbestos could not be explained by differences in fiber dimensions or surface areas, indicating an important role of mineralogical composition rather than size of fibers. PMID- 9356318 TI - Determination of parameters responsible for pharmacokinetic behavior of TCDD in female Sprague-Dawley rats. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is the most toxic member of a class of planar and halogenated chemicals. Improvements in exposure assessment of TCDD require scientific information on the distribution of TCDD in target tissues and cellular responses induced by TCDD. Since 1980, several physiologically based pharmacokinetic (PBPK) models for TCDD and related compounds have been reported. Some of these models incorporated the induction of a hepatic binding protein in response to interactions of TCDD, the Ah receptor, and DNA binding sites and described the TCDD disposition in a biological system for certain data sets. Due to the limitations of the available experimental data, different values for the same physical parameters of these models were obtained from the different studies. The inconsistencies of the parameter values limit the application of PBPK models to risk assessment. Therefore, further refinement of previous models is necessary. This paper develops an improved PBPK model to describe TCDD disposition in eight target tissues. The interaction of TCDD with the Ah receptor and with hepatic inducible CYP1A2 were also incorporated into the model. This model accurately described the time course distribution of TCDD following a single oral dose of 10 microg/kg, as well as the TCDD concentration on Day 3 after six different doses, 0.01, 0.1, 0.3, 1, 10, and 30 microg TCDD/kg, in target tissues. This study extends previous TCDD models by illustrating the validity and the limitation of the model and providing further confirmation of the potential PBPK model for us in optimal experimental design and extrapolation across doses and routes of exposure. In addition, this study demonstrated some critical issues in PBPK modeling. PMID- 9356320 TI - Stochastic Dispersal Processes in Plant Populations AB - A dispersal model for airborne pollen based on assumptions about wind directionality, gravity, and a wind threshold at which pollen is taken by the wind is developed, using a three dimensional diffusion approximation. The bivariate probability distribution of pollen receipt by flowers at the same height as the pollen source is derived. Gravity, vertical random movements, and vegetation density turn out to have similar effects on this distribution. Maximum likelihood methods for estimating the combined parameters from data with multiple point or continuous pollen sources, and one or more plant varieties, are developed. Using an example data set from the literature, it is shown that our model gives a better fit than more traditional descriptive dispersal models of the form e-ar b. We also show that estimates of important properties of the dispersal distribution, such as the variances, become considerably smaller using our model than for the more traditional models. Finally, we discuss potential extensions and evolutionary implications of these types of models. Copyright 1997 Academic Press PMID- 9356321 TI - Coupling and the Non-degeneracy of the Limit in Some Plasmid Reproduction Models AB - We settle a problem raised by Seneta and Tavare concerning the normalized limits of several plasmid reproduction models. We also show that for countably infinitely many type branching processes the extinction probability and the mass at the origin of the supercritical limit need not be the same, in contrast to the finite type case. Copyright 1997 Academic Press PMID- 9356322 TI - Density-Dependent Patch Exploitation and Acquisition of Environmental Information AB - We study density-dependent resource harvest patterns due to Bayesian foraging for different distributions of resources. We first consider a forager with information about the stochastic properties of its environment. In this case we show that when the number of food items per patch follows a distribution from the exponential family, the density dependence is given by the ratio sigma2/μ of the distribution of number of food items per patch. Bayesian foraging can therefore lead to positive (negative binomial distribution) or negative (binomial distribution) density dependent resource harvest and even to density independent (Poisson distribution) resource harvest, depending on the distribution of resources in the environment. In a second stage we incorporate learning about the distribution of resources in the whole environment. The mean of the distribution of number of food items per patch of a given environment is learnt faster than the variance of the distribution. Learning occurs faster in poorer than richer environments. Copyright 1997 Academic Press PMID- 9356323 TI - A Markov Chain Model of Coalescence with Recombination AB - Trees that describe the ancestry of DNA sequences sampled from a population may differ between loci because of genetic recombination. We seek to understand the relationship between such trees for loci that are linked with non-zero recombination rate. We consider a coalescent process model with recombination, as described by Hudson (1983; 1990). For two loci and a sample size of two sequences, a detailed analysis of this process yields the joint distribution of the two trees (one at each locus). A number of interesting results follow from this analysis, including the distribution of the number of recombination events in the history of the sample. For the general case of m loci and samples of size n, we describe an algorithm for simulating the tree building process. Because analytic results are difficult to obtain in this case, we use simulation to study properties of trees at multiple linked loci such as total tree time and number of recombination events. Copyright 1997 Academic Press PMID- 9356324 TI - Revisiting Strategic Models of Evolution: The Concept of Neighborhood Invader Strategies AB - In game-theoretic or strategic models of species evolution, the phenotype of individual organisms in a population are regarded as alternate strategies for playing a competitive game. The evolutionary outcome is predicted to conform to the "solution" of that game. The most usual solution concept adopted for the evolutionary game is that of Maynard Smith, the so-called "evolutionary stable strategies" (ESS). In this paper we explore an alternative solution concept. We call it neighborhood invader strategy (NIS). A NIS is a phenotype which is capable of invading all established populations of its neighbors. This phenotype need not be, at the same time, an ESS; and the reverse is true as well. We shall analyze this concept for a single species whose evolutionary-possibility set is a one-dimensional continuum. Copyright 1997 Academic Press PMID- 9356325 TI - The Effects of a Limited Memory Capacity on Foraging Behavior AB - We introduce a simple mathematical model to describe the behavior of a forager with a limited memory capacity in the presence of two prey types that differ in their energetic values. The model is used to analyze the effect of memory capacity on foraging behavior and on foraging energetics and it contrasts with classic optimal foraging theory, which implicitly assumes that the forager has an infinite memory capacity. Classic optimal foraging theory dictates that the low value prey type should be invariably excluded from the diet when the high value type exceeds some critical relative frequency and invariably consumed otherwise. Our model forager behaves similarly except that, as its memory capacity declines, it is increasingly predisposed to consume the low value prey when it is suboptimal to do so. Nevertheless, our analysis indicates that the energetic efficiency of a forager with an infinite memory capacity can be approximated by a forager with a memory of modest capacity, perhaps one that contains information about 5-20 previously consumed prey items. In our model, memory constraints necessarily result in so-called "partial preferences" being exercised towards the low value prey type. Thus, limited memory capacity offers a possible explanation for observed violations of the "zero-one" rule of optimal diet composition. We also find a resemblance to a sigmoidal shape in the relationship between the representation of each prey type in the diet and its relative frequency. Thus, the memory-constrained forager exhibits behavior that resembles the type III functional response. Therefore, it is possible that memory constraints such as we envisage can contribute to the explanation of both partial preferences and functional responses. Copyright 1997 Academic Press PMID- 9356326 TI - Extinction risk in a temporally correlated fluctuating environment. AB - Usually extinction risk due to environmental stochasticity is estimated under the assumption of white environmental noise. This holds for a sufficiently short correlation time tauc of the fluctuations compared to the internal time scale of population growth r-1 (tauc/r-1<<1). Using a time-discrete simulation model we investigate when the white noise approximation is misleading. Environmental fluctuations are related to fluctuations of the birth and death rates of the species and the temporal correlation of these fluctuations (coloured noise) is described by a first-order autoregressive process. We found that extinction risk increases rapidly with correlation time tauc if the strength of noise is large. In this case the white noise approximation underestimates extinction risk essentially unless temporal correlation is very small (tauc/r-1<<0.1). Extinction risk increases only slowly with correlation time if the strength of noise is small. Then the white noise approximation may be used even for stronger temporal correlations (tauc/r-1>/=0.1). Thus, the estimation of extinction risk on the basis of white or coloured noise must be justified by time scale and strength of the fluctuations. Especially for species that are sensitive to environmental fluctuations the applicability of the white noise approximation should be carefully checked. PMID- 9356327 TI - Epidemic thresholds and vaccination in a lattice model of disease spread. AB - We use a lattice-based epidemic model to study the spatial and temporal rates of disease spread in a spatially distributed host population. The prevalence of the disease in the population is studied as well as the spread of infection about a point source of infection. In particular, two distinct critical population densities are identified. The first relates to the minimum population density for a epidemic to occur, whilst the second is the minimum population density for long term persistence to occur. Vaccination regimes are introduced that are used to measure the impact of spatially and nonspatially dependent intervention strategies. Specifically we show how a ring of vaccinated susceptibles, of sufficient thickness, can halt the spread of infection across space. PMID- 9356328 TI - Fisher's fundamental theorem of natural selection revisited. AB - W. J. Ewens, following G. R. Price, has stressed that Fisher's fundamental theorem of natural selection about the increase in mean fitness is of general validity without any restrictive assumptions on the mating system, the fitness parameters, or the numbers of loci and alleles involved, but that it concerns only a partial change in mean fitness. This partial change is obtained by replacing the actual genotypic fitnesses by the corresponding additive genetic values and by keeping these values fixed in the change of the mean with respect to changes in genotype frequencies. We propose an alternate interpretation for this partial change which uses partial changes in genotype frequencies directly consequent on changes in gene frequencies, the fitness parameters being kept constant. We argue that this interpretation agrees more closely with Fisher's own explanations. Moreover, this approach leads to a decomposition for the total change in mean fitness which explains, unifies, and extends previous decompositions. We consider a wide range of models, from discrete-time selection models with nonoverlapping generations to continuous-time models with overlapping generations and age effects on viability and fecundity, which is the original framework for Fisher's fundamental theorem. PMID- 9356329 TI - Linkage disequilibrium and the infinitesimal limit. AB - Under the classical Fisher-Bulmer infinitesimal model of quantitative genetics, the within-family distribution for an additive trait with no environmental component is Gaussian with mean at the mid-parent value and a variance which is the same for all families. When an additive trait is determined by unlinked loci, the Fisher-Bulmer model can arise in the limit as the number of loci contributing to variation in the trait increases. However, a counterexample is presented where the Fisher-Bulmer model fails to arise in the infinite locus limit because there is too much linkage disequilibrium. An example is also presented where a degenerate form of the Fisher-Bulmer model arises. Under what conditions does the Fisher-Bulmer model arise in the infinite locus limit? It follows from the central limit theorem that the within-family distribution is Gaussian. But, under what conditions is the within-family distribution the same for almost all families in the population? An alternative population genetic derivation of the Fisher-Bulmer model is presented for a population at linkage equilibrium. This approach is then extended to allow many patterns of linkage disequilibrium. Diallelic models are used to illustrate the type of linkage disequilibrium allowed. The results on the limiting behaviour of population genetic models with many unlinked loci can be regarded as special cases of a more general limiting property of sequences of random variables. A possible application of this more general result to models of cultural inheritance is suggested. PMID- 9356331 TI - Integration of visna virus DNA occurs and may be necessary for productive infection. AB - Proviral integration is thought to be an obligate step of the retroviral replication cycle but the lentivirus visna has been reported to replicate in sheep choroid plexus (SCP) cultures in the absence of proviral integration. Because of new evidence that visna virus has a functional integrase, we reexamined visna virus infection of SCP cultures and found that proviral integration does indeed occur in this setting. While the majority of viral DNA remains unintegrated, integrated proviruses arise early in infection and accumulate over time. The sequences of the resulting host-virus DNA junctions show that, like other retroviruses, visna loses terminal nucleotides from its DNA upon integration. However, unlike other retroviruses, in over half the host-U3 junctions analyzed only a single nucleotide was lost such that the universally conserved CA dinucleotide, two nucleotides from the end of unintegrated viral DNA, did not directly abut host sequences in the provirus. We analyzed the role of integration in visna replication by introducing a series of five mutations into the integrase gene of molecularly cloned visna virus LV1-1KS1. Each mutation abolished viral replication, suggesting that integration may be an obligatory step in replication. We also documented productive infection of SCP cultures in which cell division had been blocked by g-irradiation. The ability of visna to integrate and to replicate in nondividing cells points to the possible utility of visna-based vectors for gene transfer into differentiated cells. PMID- 9356330 TI - The dynamics of measles epidemics. AB - The interepidemic interval (T) of measles in London from 1647 to 1837 evolved progressively from 5-yearly to 2-yearly by 1800. Measles mortality was significantly ( p<0.001) cross-correlated with the annual wheat prices, a good index of nutrition although at a 2-year lag. Epidemics correlated with low autumn temperatures (p<0. 001). A linearised model of the dynamics of epidemics shows that T is determined by the product of population (N) and susceptibility (beta) and that the system will settle at its steady state unless the epidemics are driven. It is suggested that (i) the progressive change in T was caused by a rise in population size (N) and an increased susceptibility (beta) related to malnutrition and (ii) epidemics were driven by oscillations in low autumn temperature (p<0. 001) and by cycles of susceptible young children produced by malnutrition during pregnancy. PMID- 9356332 TI - Reconstitution of a functional bovine papillomavirus type 1 origin of replication reveals a modular tripartite replicon with an essential AT-rich element. AB - A functional replication origin was reconstituted using oligonucleotide cassettes corresponding to three sequence subelements within the Bovine Papillomavirus Type 1 (BPV-1) replication origin: the 23-bp AT-rich region (ATR), the 18-bp binding site for the viral replication initiator protein E1 (E1BS), and a binding site for the viral transcriptional transactivator and replication enhancer protein E2 (E2BS). Replication of the reconstituted origin depended on heterologous expression of both the E1 and E2 proteins and on the presence of both the E1BS and E2BS, indicating that it is functionally analogous to the authentic BPV-1 origin. In addition, pairwise testing of subelement combinations revealed that the ATR was also essential and that a functional origin required at least one copy of all three subelements. While the E1BS and E2BS are sequence-specific elements, the function of the BPV-1 ATR could be at least partially substituted with heterologous AT-rich sequences, suggesting that the role of this element is primarily AT content-dependent rather than sequence-dependent. A stringent requirement for the ATR was also observed in the context of an authentic minimal origin sequence confirming that it is an intrinsic property of the BPV-1 origin and not simply an artifact of the reconstitution system. This study indicates that the minimal functional BPV-1 origin shares the tripartite modular organization characteristic of other simple eukaryotic replication origins. The reconstitution system described now provides a convenient approach to define the physical and functional interrelationships between the three subelements in a systematic fashion. PMID- 9356333 TI - Inducible knockout of the interleukin-2 receptor alpha chain: expression of the high-affinity IL-2 receptor is not required for the in vitro growth of HTLV-I transformed cell lines. AB - Adult T cell leukemia (ATL) is an aggressive malignancy that is associated with HTLV-I infection and characterized by constitutive expression of the high affinity interleukin-2 receptor. The alpha subunit of the high-affinity receptor (IL-2Ralpha), which is normally present only on activated T cells, is specifically upregulated by HTLV-I and constitutively expressed on fresh leukemic cells from ATL patients as well as cell lines transformed by HTLV-I in vitro. Here we directly address the functional significance of IL-2Ralpha expression in HTLV-I transformed cell lines by using an endoplasmic reticulum-targeted single chain antibody to inhibit the cell surface expression of IL-2Ralpha. Using constitutive and tetracycline-repressible systems to express the ER-targeted antibody against IL-2Ralpha, we have reduced cell surface expression of IL 2Ralpha by more that 2 logs of mean fluorescence intensity to virtually undetectable levels in the IL-2-independent HTLV-I-transformed cell lines C8166 45 and HUT102. No toxicity was associated with the intracellular retention of IL 2Ralpha, and the growth rate of the IL-2Ralpha-negative cells was in each case comparable to that of the parental cell line. We conclude that cell surface expression of IL-2Ralpha is dispensable for the in vitro growth of these HTLV-I transformed cells. PMID- 9356334 TI - Effects of domain-switching and site-directed mutagenesis on the properties and functions of the VP7 proteins of two orbiviruses. AB - Based on the crystal structure of the VP7 major core protein of bluetongue virus serotype 10 (BTV-10) and that of the top domain of the VP7 protein of African horsesickness virus serotype 4 (AHSV-4), chimeras and site-directed mutants of the proteins were constructed and the products analyzed with respect to their properties and functions. Chimeras with the central upper domain of BTV-10 VP7 replaced by that of AHSV-4 VP7 (construct BAB) formed trimers, as did the converse construct (ABA). Further, both proteins exhibited the expected conformational epitopes of the constituent sequences. Using BAB VP7 it was demonstrated that residues of the upper domain of AHSV-4 VP7 contribute to the observed insolubility of the protein. By contrast, ABA VP7 protein was as soluble as wild-type BTV-10 VP7. Replacement of selected amino acid residues in the top domain (e.g., A167 by R; F209 by T) improved the solubility of BAB VP7. Since the trimeric BAB and ABA VP7 proteins did not form core-like particles (CLPs) when coexpressed with BTV VP3, it was concluded that trimerization of chimeric VP7 is not sufficient for CLP formation. When the N-terminal region of the ABA protein (aa 1-120) was replaced by the respective sequences of BTV VP7 (construct BBA), the protein aggregated and did not form CLPs with coexpressed BTV VP3, most likely due to disruption of the required contacts between the N- and C-terminal regions of the bottom domain, leading to incorrect folding of the chimera. PMID- 9356335 TI - Determination of the minimal amount of Tat activity required for human immunodeficiency virus type 1 replication. AB - The Tat protein of human immunodeficiency virus type 1 (HIV-1) is a potent trans activator of transcription from the viral LTR promoter. Previous mutagenesis studies have identified domains within Tat responsible for binding to its TAR RNA target and for transcriptional activation. The minimal Tat activation domain is composed of the N-terminal 48 residues, and mutational analyses identified a cluster of critical cysteines. The importance of four highly conserved aromatic amino acids within the activation domain has not been thoroughly investigated. We have systematically substituted these aromatic residues (Y26, F32, F38, Y47) of the HIV-1 LAI Tat protein with other aromatic residues (conservative mutation) or alanine (nonconservative mutation). The activity of the mutant Tat constructs was measured in different cell lines by transfection with a LTR-CAT reporter plasmid. The range of transcriptional activities measured for this set of Tat mutants allowed careful assessment of the level of Tat activity required for optimal viral replication. To test this, the mutant Tat genes were introduced into the pLAI infectious molecular clone and tested for their effect on virus replication in a T-cell line. We found that a twofold reduction in Tat activity already affects viral replication, and no virus replication was measured for Tat mutants with less than 15% activity. This strict correlation between Tat activity and viral replication demonstrates the importance of the Tat function to viral fitness. Interestingly, a less pronounced replication defect was observed in primary cell types. This finding may correlate with the frequent detection of proviruses with Tat-inactivating mutations in clinical samples. PMID- 9356336 TI - Characterization of cucumber mosaic virus. IV. Movement protein and coat protein are both essential for cell-to-cell movement of cucumber mosaic virus. AB - cDNA clones of cucumber mosaic virus (CMV) RNA 3 were modified to express the jellyfish green fluorescent protein (GFP) in place of the 3a movement protein (MP) or coat protein (CP), as fusions to the N (GFP-3a) or C (3a-GFP) terminus of the MP or from a separate open reading frame as part of tricistronic RNAs 3. CMV RNA transcripts containing the individual modified RNAs 3 were unable to infect either Nicotiana tabacum or Nicotiana benthamiana systemically. Infection, as measured by confocal microscopy of GFP fluorescence, generally was limited to one to three epidermal cells at each inoculation site. Limited cell-to-cell movement, but not systemic movement, could be detected by complementation involving expression of MP and CP from two different RNA 3 constructs, each also expressing GFP. Infection involving RNA 3 expressing the GFP-3a fusion showed bright granules of variable size distributed predominantly and nonuniformly throughout the cytoplasm and, to a lesser extent, associated with the cell wall in single fluorescent cells, while infections expressing the 3a-GFP fusion showed bright, punctate fluorescence associated only with the cell wall. Infected cells expressing either 3a-GFP or free GFP showed a halo of less bright, fluorescent, neighboring cells, indicating limited movement of GFP. The initially infected cells also allowed movement of 10-kDa fluorescent dextran to the neighboring halo cells, while infection did not spread, suggesting different requirements for movement of either MP or dextran versus RNA. PMID- 9356337 TI - Recovery of infectious SV5 from cloned DNA and expression of a foreign gene. AB - A complete cDNA clone of the genome (15,246 nucleotides) of the paramyxovirus SV5 was constructed from cDNAs such that an anti-genome RNA could be transcribed by T7 RNA polymerase and the correct 3' end generated by cleavage using hepatitis delta virus ribozyme. The plasmid encoding the antigenome sequence was transfected into cells previously infected with recombinant vaccinia virus that expressed T7 RNA polymerase, together with helper plasmids that expressed the viral replication proteins, NP, P, and L, under the control of the T7 polymerase promoter. Rescue of the RNA genome from DNA was demonstrated by recovering SV5 with the tag restriction sites introduced into the DNA clone, using RT-PCR of the genome RNA and nucleotide sequencing. Rescue of SV5 from DNA did not require expression of the viral V protein as a helper plasmid, suggesting that V protein is not essential for initial replication. The infectious cDNA of SV5 was also manipulated to express green fluorescent protein (GFP) under the control of SV5 transcriptional start and stop signals introduced between the HN and L genes. The amount of GFP that was expressed varied depending on the nature of the newly introduced transcription signals. PMID- 9356338 TI - Provirus variants of the bovine leukemia virus and their relation to the serological status of naturally infected cattle. AB - Infection of cattle with the bovine leukemia virus (BLV) results in a strong permanent antibody response to the BLV antigens some weeks after infection. However, cattle may carry provirus and not have detectable antibody titers. To prove the occurrence of different BLV provirus variants in German cattle and to study the influence of special BLV variants on the immunoreaction, a 444-bp fragment of the env gene of 35 naturally BLV infected animals was analyzed. Seven different groups of BLV provirus variants were found on the basis of restriction fragment length polymorphism. Three BLV provirus variant groups and five additionally sequenced BLV isolates showed a high similarity to BLV provirus isolates from other geographical areas. The variation in nucleotide sequence of the five BLV isolates compared with nine previously sequenced BLV isolates ranged up to 5. 3%. While BLV provirus variant groups A, C, D, E, F, and G were clearly related to agar-gel immunodiffusion test (AGID)- and enzyme-linked immunosorbent assay (ELISA)-positive animals, BLV provirus variant group B was solely found in permanent AGID- and ELISA-negative or in transient ELISA-positive animals. Altogether, these results indicate that special BLV provirus variants may be responsible for atypical forms of BLV infection in cattle. PMID- 9356339 TI - Isolation of RNA aptamers specific to the NS3 protein of hepatitis C virus from a pool of completely random RNA. AB - Hepatitis C virus (HCV) is a single-stranded RNA virus and its genome is translated into a single large polyprotein. The viral-encoded NS3 protein possesses protease, nucleoside triphosphatase, and helicase activities. Since these activities appear to be important for viral replication, efforts are being made to identify compounds that might inhibit the enzymatic activities of NS3 and serve as potential anti-HCV agents. We used a genetic selection strategy in vitro to isolate, from a pool of completely random RNA (120 random bases), those RNA aptamers that could bind to NS3. After six cycles of selection and amplification, 14% of the pooled RNAs could bind specifically to the NS3 protein. When the aptamers in the pool (cycle 6) were analyzed for binding and inhibition of the proteolytic activity of NS3 with the NS5A/NS5B peptide as substrate (S1), two aptamers, designated G6-16 and G6-19 RNA, were found to inhibit NS3 in vitro. Kinetic studies of the inhibition revealed that the aptamer G6-16 inhibited the NS3 protease with an inhibitory constant (Ki) of 3 microM. We also analyzed aptamers G6-16 and G6-19 for their action with a longer protein substrate (amino acid region 2203-2506) and found that these aptamers efficiently inhibited the proteolytic activity of NS3. In addition, both G6-16 and G6-19 aptamers were found to inhibit the helicase activity of NS3. Since these aptamers possesses dual inhibitory function for NS3, they could prove to be useful as anti-HCV drug leads. PMID- 9356340 TI - Capsid protein and helper component-proteinase function as potyvirus cell-to-cell movement proteins. AB - The role of bean common mosaic necrosis potyvirus (BCMNV) and lettuce mosaic potyvirus (LMV) proteins was investigated in terms of their capacity to function as viral movement proteins (MPs). Using Escherichia coli-expressed proteins and microinjection techniques, direct evidence was obtained that both the potyviral capsid protein (CP) and helper component- proteinase (HC-Pro) function in this capacity, in that both proteins (a) trafficked from cell to cell, (b) induced an increase in plasmodesmal size exclusion limit, and (c) facilitated cell-to-cell movement of viral RNA. CP and HC-Pro mutants were also produced and used in microinjection experiments. Mutations in the core region of the CP either impaired (single and double amino acid substitution mutants) or abolished (triple amino acid substitution mutant) cell-to-cell movement, as did C-terminal deletion mutants in HC-Pro. The BCMNV P1, CI, NIa, and NIb proteins did not exhibit viral MP properties, but NIa and NIb proteins were found to accumulate within the nuclei of injected cells. These results further establish the multifunctional nature of the potyvirus CP and HC-Pro. PMID- 9356341 TI - Inhibition of serum- and calcium-induced differentiation of human keratinocytes by HPV16 E6 oncoprotein: role of p53 inactivation. AB - We have recently shown that human papillomavirus (HPV16) E6 oncoprotein exhibits two separate biological activities in genital keratinocytes (PHKs). E6 protein by itself is capable of inducing colonies of proliferating cells resistant to serum and calcium-induced differentiation, whereas both E6 and E7 are required for immortalization of PHK. Using epitope-tagged E6 carboxy-terminal truncation mutants, we mapped the domain between amino acid residues 132 and 141 as being essential for the induction of differentiation resistance (L. Sherman and R. Schlegel, J. Virol. 70, 3269-3279, 1996). To determine whether E6 protein's ability to alter PHK response to serum and calcium was associated with its ability to inactivate p53, we evaluated each of the above E6 mutants and three E6 natural variants in these respective assays. Our results demonstrate that the E6 region spanning residues 132-141 is required for p53 degradation and for abrogation of p53 transactivation, suggesting a possible correlation between E6 biological activity in altering differentiation and loss of p53 function. To evaluate whether selective inactivation of p53 is sufficient for altering the response of PHK to serum and calcium we investigated the capacity of plasmids encoding a dominant mutant human p53 and human MDM-2 to functionally substitute for E6 in colony formation in PHK. Plasmids were verified for their ability to inactivate wild-type p53 by testing their capacity to abrogate the p53 transactivation function. The results obtained showed that vectors encoding human MDM-2 and mutant p53, while active in inhibition of p53-dependent transactivation and capable of expressing stable proteins in PHK, failed to induce colonies of proliferating cells resistant to serum and calcium differentiation. These data argue that p53 inactivation may not be the sole E6 function required for altering the response of PHK to serum- and calcium-triggered differentiation. PMID- 9356342 TI - North American and French caprine arthritis-encephalitis viruses emerge from ovine maedi-visna viruses. AB - The full extent of genetic diversity among small ruminant lentiviruses (SRLVs), i.e., caprine arthritis encephalitis viruses (CAEVs) and maedi-visna viruses (MVVs), remains unknown. This is due in part to the fact that few sequences of CAEV are available. To contribute to this knowledge, gag, pol, and env nucleotide sequences from an SRLV named CA680 originating from a goat from western France were determined. This analysis revealed that this virus is closely related to the Cork and 63 CAEV American isolates. Mismatched amino acids between the CA680 virus and prototype CAEVs ranged from 6.7, 0. 7, and 17.5% for gag, pol, and SU sequences, respectively. The differences between the CA680 virus and MVV prototypes ranged from 16.5, 12.5, and 32.3% for the protein sequences, respectively. A screening using a heteroduplex mobility assay (HMA) adapted to SRLVs revealed that 6 of 10 caprine virus field isolates were closely related to CA680, indicating that this latter isolate was a prototype of CAEVs common in the west of France. Phylogenetic trees drawn using CA, RT, or SU sequences of numerous SRLVs and rooted with EIAV sequences revealed that CA680 and CAEV prototypes, all infectious for goat, clustered in one group. From these HMA and phylogenetic analyses, it appears that U.S. and French caprine SRLVs form a clade that had emerged from a much more diverse group containing all SRLVs infectious for sheep. These ovine SRLVs form a more ancient group in which the EIAV is rooted. PMID- 9356343 TI - Definition of the primary structure of hepatitis B virus (HBV) pre-S hepatocyte binding domain using random peptide libraries. AB - The pre-S-specific monoclonal antibody MA 18/7 has been shown to inhibit the binding of HBV to HepG2 cells and liver membranes. This antibody can thus be used to identify the critical residues of the pre-S region involved in the hepatocyte binding domain. Using overlapping 7-mer peptides representing the pre-S region of HBV, the epitope recognized by MA 18/7 was shown to contain sequences from both the pre-S1 and pre-S2 regions, thus indicating that the hepatocyte-binding domain is conformationally dependent. To further characterize the primary structure of the hepatocyte-binding domain on the pre-S protein, a phage-displayed 15-mer peptide library and a 8-mer solid phase peptide library were used to analyze the fine specificity of the monoclonal antibody MA 18/7. Several mimotopes were identified with the phage-displayed peptide library, the majority of which possess a central motif with at least three identical residues present within the native pre-S1 sequence. No significant consensus sequences were found when these mimotopes were compared to the pre-S2 sequence. Mimotopes identified using the solid-phase peptide library also contained a similar motif. All phage mimotopes and a single mimotope from the solid-phase peptide library competed with recombinant HBsAg particles containing the pre-S1 region for binding to MA 18/7. Mouse antisera raised against four mimotopes from the phage display library reacted with HBsAg particles containing pre-S sequences. The data show that the structure of the pre-S molecule around the conserved DPAF motif in the pre-S region may have a functional role in binding HBV to cellular receptors, and that the central motif identified in mimotopes of this region may offer a novel strategy target for the improvement of existing hepatitis B vaccines which, at present, are mostly devoid of pre-S specificities. PMID- 9356344 TI - RNA binding activity of NIa proteinase of tobacco etch potyvirus. AB - The C-terminal domain of NIa protein (NIaPro) from tobacco etch potyvirus (TEV) is a sequence-specific proteinase required for processing of the viral polyprotein. This proteinase also interacts with NIb, the TEV RNA-dependent RNA polymerase. NIaPro and two NIaPro-containing polyproteins (NIa and 6/NIa) were analyzed from extracts of recombinant Escherichia coli. Using RNA-protein blot and UV-crosslinking assays, NIaPro and the NIaPro-containing polyproteins were shown to possess RNA-binding activity. NIaPro bound nonspecifically to several RNAs, including plus- and minus-strands of the TEV 5' and 3' noncoding regions. Saturation binding data obtained using the UV-crosslinking assay were consistent with a possible cooperative RNA-binding activity of NIaPro. In addition, the RNA binding activities of NIaPro and full-length NIa protein were similar. Based on its RNA-binding activity and other known functions, NIaPro or a NIaPro-containing polyprotein is proposed to serve one or more direct roles during TEV RNA synthesis. PMID- 9356345 TI - Subdivision of the pestivirus genus based on envelope glycoprotein E2. AB - Conventionally, the genus Pestivirus of the family Flaviviridae has been divided into bovine viral diarrhea virus (BVDV), classical swine fever virus (CSFV), and border disease virus (BDV). To date, BDV and BVDV have been isolated from different species, whereas CSFV seems to be restricted to swine. Pestiviruses are structurally and antigenically closely related. Envelope glycoprotein E2 is the most immunogenic and most variable protein of pestiviruses. We cloned E2 genes of many different pestivirus strains, including those from a deer and a giraffe. The E2 genes were transiently expressed, characterized with monoclonal antibodies, sequenced, and compared. Based on these data, we can delineate six major groups within the Pestivirus genus. Four groups correspond to defined genotypes, whereas the two other groups could be new genotypes within the Pestivirus genus. One group comprises CSFV strains isolated from swine. A second group consists of BDV strains Moredun, L83, and X818, which have been isolated from sheep, and strain F from swine. A third group contains strain BD78 from sheep, strain 5250 from swine, and strain 178003 from cattle. On the basis of E2, these viruses are very similar to BVDV strains associated with acute severe outbreaks of bovine viral diarrhea, so-called type 2 BVDV. The fourth group consists of BVDV strains originating predominantly from cattle. This BVDV group can be divided into two subtypes or subgroups BVDV Ia and Ib: BVDV Ia contains viruses from the United States, such as like NADL and Oregon, and some others, such as 150022 and 1138 from Europe. Subgroup BVDV Ib contains strain Osloss and several Dutch isolates. The fifth and sixth "groups" could be proposed as two new genotypes and contain strains Deer and Giraffe, respectively. PMID- 9356346 TI - Characterization of new simian foamy viruses from African nonhuman primates. AB - Simian foamy viruses (SFV) are exogenous retroviruses present in most if not all nonhuman primate species. Baboons and other African monkey species are known to harbor SFVs, yet there is presently no data in regard to their genetic relationship. Here we studied SFVs from baboons as compared to other SFVs isolated from a Hamlyn's guenon, a patas monkey, and a vervet. By Western blot analysis, the gag precursor proteins (p74/p70) were detected from all SFVs. In addition, the envelope glycoproteins from a vervet isolate (SFV-Agm2) were comparable in size to the env precursor gp130, the exterior glycoprotein (gp70), and the transmembrane protein (gp48) as detected by lentil lectin binding and radioimmunoprecipitation (RIPA). Molecular comparison of PCR amplified products from pol and LTR regions of each SFV demonstrated a close relationship among baboon SFVs while SFVs from patas, Hamlyn's guenon, and vervet clustered together. The baboon viruses only varied by 4% among each other in the LTR region; however, as much as 26% variation was noted when compared to the other African monkey SFVs. To determine the prevalence rate of SFV-Bab in our baboon colony, we employed both Western blotting and PCR analysis. Antibodies to SFV gag precursor proteins were seen in 7 of 10 infants; however, none were positive by PCR, suggesting that these infants were virus negative and that their antibodies were maternal in origin. Only one juvenile (1/10) and all adults (38/38) were infected with SFV. Taken together these results suggest that SFVs have arisen and diverged along with the evolution of their natural hosts. Furthermore, the high prevalence rates to SFV seen in adult baboons strongly suggest a sexual or oral routes of transmission. PMID- 9356348 TI - Identification of a consensus mutation in M protein of vesicular stomatitis virus from persistently infected cells that affects inhibition of host-directed gene expression. AB - In addition to its function in virus assembly, the viral matrix (M) protein of vesicular stomatitis virus (VSV) inhibits host-directed gene expression. The goal of this study was to determine whether sequence changes in M protein contribute to a reduced shut off of host gene expression in cells persistently infected with VSV. Viruses isolated from L cells persistently infected with VSV inhibited host RNA synthesis more slowly than wild-type (wt) VSV. M genes of the persistent viral population were cloned and sequenced. One mutation, an N to D change at position 163 of the protein sequence (N163D), was common to all the molecular clones. The N163D M protein was synthesized from transfected mRNA at a rate that was 30% of that of wt M protein, but was turned over at a rate that was similar to that of wt M protein. Transfection of mRNA encoding N163D M protein inhibited expression of a cotransfected target gene encoding chloramphenicol acetyl transferase (CAT), but the inhibition was 6 to 10 times less effective than transfection of equivalent amounts of wt M mRNA. This difference could not be accounted for by differences in translation of CAT mRNA. Thus, when the differences in M protein expression were taken into account, N163D M protein was 2 to 3 times less effective than wt M protein in the inhibition of host-directed gene expression, similar to the differences in host transcription observed in virus-infected cells. Point mutations in addition to the N163D mutation were found in about half of the M gene molecular clones. The M gene of an independently isolated molecular clone, N163D.2, contained two additional point mutations in its carboxy terminal region. N163D.2 M protein was highly defective in inhibition of host gene expression and was turned over more rapidly than wt M protein. These results support the idea that M gene mutations contribute to a reduced cytopathic effect in cells persistently infected with VSV. PMID- 9356347 TI - Analysis of 74 kb of DNA located at the right end of the 330-kb chlorella virus PBCV-1 genome. AB - This report completes a preliminary analysis of the sequence of the 330,740-bp chlorella virus PBCV-1 genome, the largest virus genome to be sequenced to date. The PBCV-1 genome is 57% the size of the genome from the smallest self replicating organism, Mycoplasma genitalium. Analysis of 74 kb of newly sequenced DNA, from the right terminus of the PBCV-1 genome, revealed 153 open reading frames (ORFs) of 65 codons or longer. Eighty-five of these ORFs, which are evenly distributed on both strands of the DNA, were considered major ORFs. Fifty-nine of the major ORFs were separated by less than 100 bp. The largest intergenic distance was 729 bp, which occurred between two ORFs located in the 2.2-kb inverted terminal repeat region of the PBCV-1 genome. Twenty-seven of the 85 major ORFs resemble proteins in databases, including the large subunit of ribonucleotide diphosphate reductase, ATP-dependent DNA ligase, type II DNA topoisomerase, a helicase, histidine decarboxylase, dCMP deaminase, dUTP pyrophosphatase, proliferating cell nuclear antigen, a transposase, fungal translation elongation factor 3 (EF-3), UDP glucose dehydrogenase, a protein kinase, and an adenine DNA methyltransferase and its corresponding DNA site specific endonuclease. Seventeen of the 153 ORFs resembled other PBCV-1 ORFs, suggesting that they represent either gene duplications or gene families. PMID- 9356349 TI - Screening human tumor samples with a broad-spectrum polymerase chain reaction method for the detection of polyomaviruses. AB - Polyomaviruses induce tumors of different histological types when inoculated into experimental animals. An etiological role for this virus group in the development of malignant tumors in humans remains questionable, despite several reports demonstrating the presence of SV40, JCV, and BKV DNA in human cancers. Only two human polyomavirus types are known to date: JCV, causing progressive multifocal leukoencephalopathy (PML) under severe immunosuppression, and BKV, first isolated from the urine of a renal transplant recipient and associated with hemorrhagic cystitis. We developed a degenerate polymerase chain reaction assay in an attempt to identify additional, presently unknown human polyomavirus types that may be involved in the malignant transformation of human tissues. A large part of the gene coding for the viral capsid protein VP1 is highly conserved in nine polyomavirus types (and their strains) and was therefore selected as most suitable for the primer design. Degenerate oligonucleotide primers were deduced from four different conserved amino acid motifs in this region. Three different sets of primers were included in each test to obtain the highest sensitivity in combination with primers with the lowest degeneracy numbers. The sensitivity obtained ranged from 1 copy/cell for bovine polyomavirus to 100 copies/cell for LPV after ethidium bromide staining and was increased at least 10-fold after hybridization with a radiolabeled probe. A subsequent seminested amplification allowed for the detection of 1 copy/cell for LPV. These degenerate primers were applied to analyze bladder carcinomas, Hodgkin lymphomas, meningiomas, Kaposi tumors, and Kaposi-derived cell lines. No polyomavirus DNA sequences could be detected. PMID- 9356350 TI - Functional conservation of HTLV-1 rex balances the immune pressure for sequence variation in the rex gene. AB - Naturally occurring mutations in Human T-cell Leukemia Virus Type 1 (HTLV-1) Tax protein lead to loss of recognition by cytotoxic T-lymphocytes. Most of these mutations also abolish or severely impair the transactivation function of Tax. Ninety percent of the rex gene, which encodes the viral regulator of mRNA splicing (Rex), overlaps with the tax gene. In this paper, we report that four previously described point mutations in tax that abolished CTL recognition and activity did not alter either the dimerisation function or the ability to export viral mRNA of the corresponding Rex proteins. Rex proteins containing two other amino acid changes were likewise functional. However, five Rex deletion mutants, predominantly but not exclusively found in HAM/TSP patients, had all lost these functions. We conclude that, although the Tax protein is subject to strong CTL mediated selection, there are stronger functional constraints on amino acid variation in Rex. This may limit the variation in the tax/rex nucleotide sequence which results in immune evasion. PMID- 9356351 TI - p53/E1b58kDa complex regulates adenovirus replication. AB - We have explored a role for the adenovirus (Ad5) E1b58kDa/p53 protein complex in adenovirus replication. This was done by using virus mutants containing different defects in the E1b58kDa gene and cell lines that express either a wild-type p53 protein or a mutant p53 protein. We find that infection of wild-type p53 containing cells with wild-type Ad5 causes a shutoff of p53 and alpha-actin protein synthesis by distinct mechanisms, but neither occurs in mutant p53 cells. Our data also indicate that the shutoff is dependent on formation of the p53/E1b complex and may also involve another virus protein, E4ORF6. Following from these observations we asked whether failure to form the complex resulted in impaired adenovirus replication. Our experiments showed that neither wild-type Ad5 nor the E1b mutant dl338 could replicate in cells expressing a mutant p53 protein, but that wild-type adenovirus replicated well in wild-type p53-expressing cells. Collectively, our data suggest that the interaction between p53 and the E1b58kDa protein is necessary for efficient adenovirus replication. This is the first time such a direct link between the complex and virus replication has been demonstrated. These data raise serious questions about the usefulness of E1b defective viruses in tumor therapy. PMID- 9356352 TI - Self-assembly and protein-protein interactions between the SV40 capsid proteins produced in insect cells. AB - Soluble SV40 capsid proteins were obtained by expression of the three late genes, VP1, VP2, and VP3, in Sf9 cells using baculovirus expression vectors. Coproduction of the capsid proteins VP1, VP2, and VP3 was achieved by infecting Sf9 cells with the three recombinant baculovirus species at equal multiplicities. All three proteins were found to be localized in the nuclear fraction. Electron microscopy of nuclear extracts of the infected cells showed an abundance of SV40 like capsid structures and heterogeneous aggregates of variable size, mostly 20 45 nm. Under the same staining conditions wild-type SV40 virions are 45 nm. The capsid-like particles sedimented in glycerol gradients similarly to authentic wild-type SV40 virions. Pentamers of the major capsid protein VP1 were also seen. Protein analysis on sucrose gradients demonstrated that the capsid-like particles can be disrupted by treatment with the reducing agent dithiothreitol and the calcium chelator EGTA. The capsid-like particles were found to be significantly less stable than SV40 virions and were partially stabilized by calcium ions. Understanding the complex interactions between the capsid proteins is important for the development of an efficient in vitro packaging system for SV40 virions and pseudovirions. PMID- 9356353 TI - TNFalpha cooperates with the protein kinase A pathway to synergistically increase HIV-1 LTR transcription via downstream TRE-like cAMP response elements. AB - Activating protein-1 (AP-1) binding TPA responsive elements (TRE) are located downstream of the transcription initiation site in the U5 region of the HIV-1 long terminal repeat (LTR). These downstream sequence elements, termed DSE, can bind both AP-1 and CREB/ATF transcription factors. Recently, we demonstrated that the DSE are also cAMP-responsive elements (CRE), since they mediated activation signals elicited by cholera toxin (Ctx), a potent activator of the cAMP-dependent protein kinase A (PKA) signal transduction pathway. In the present study, we demonstrate that the HIV-1 DSE can mediate the transcriptional synergy elicited by the combination of Ctx and TNFalpha. Ctx combined with TNFalpha or IL-1beta to produce a synergistic increase in p24 antigen production in U1 promonocytic cells. Transfection studies of LTR reporter constructs indicated that mutation of the DSE sites abrogated the LTR-mediated synergy induced by Ctx and TNFalpha, whereas the synergy induced by Ctx and IL-1beta was unaffected, suggesting TNFalpha and IL-1beta cooperate differently with the cAMP/PKA activation pathway to induce HIV-1 expression in U1 cells. Because the DSE are also TRE sites, we assessed the effect of the agonist combinations on AP-1-dependent transcription. TNFalpha as well as IL-1beta cooperated with Ctx to produce a synergistic activation of AP-1-mediated transcription. These data indicate that the TRE-like cAMP-responsive DSE sites within the 5'-untranslated leader can mediate the transcriptional cooperativity between TNFalpha and the cAMP/PKA pathway. Since the DSE and TRE sites cannot bind CREB/ATF homodimers, we propose a mechanism in which the HIV-1 DSE bind heterodimers composed of both AP-1 and CREB/ATF proteins. PMID- 9356354 TI - Productive penetration of rotavirus in cultured cells induces coentry of the translation inhibitor alpha-sarcin. AB - Internalization of rotavirus in MA104 cells was found to induce coentry of alpha sarcin, a toxin that inhibits translation in cell-free systems and to which cells are normally impermeable. Entry of the toxin, measured by inhibition of protein synthesis at early times after infection, correlated with virus penetration leading to expression of infectivity, since toxin entry (1) was induced only by trypsin-treated triple-layered virions, to a degree dependent on the toxin and the virus concentration; (2) correlated with the degree of permissivity of different cell lines to rotavirus infection; (3) was inhibited to a similar extent as infectivity by treatment of cells with neuraminidase; and (4) was inhibited by pre- or postadsorption incubation of the virus with neutralizing monoclonal antibodies to VP7 and VP4 (VP8*). Neither the virus infectivity nor the toxin coentry was significantly affected by treatment of cells with bafilomycin A1, an inhibitor of the vacuolar proton ATPase, indicating that both events are independent of the endosomal acid pH. Virus-like particles (VLP), composed of rotavirus proteins 2/6/7/4, but not 2/6/7 or 2/6, were able to induce toxin entry as efficiently as virions. Use of genetically modified VLP in combination with the toxin coentry assay, which measures entry through a productive pathway, should allow identification of the regions of the outer capsid proteins essential for rotavirus penetration. PMID- 9356356 TI - Variations in the VPg protein allow a potyvirus to overcome va gene resistance in tobacco. AB - The va gene is used in commercial Burley tobacco cultivars including cv TN 86 to confer resistance to tobacco vein mottling virus (TVMV) and, to some extent, other potyviruses. A naturally occurring strain of TVMV (TVMV-S), which overcomes this resistance, was isolated from TN 86 plants. To investigate the mechanism by which TVMV-S overcomes va gene resistance, a cDNA clone encompassing the complete genome of TVMV-S was produced. Using chimeric transcripts combining regions of TVMV-S and regions of the wild-type strain (TVMV-WT) to which TN 86 is resistant, it was demonstrated that a domain within the VPg protein is responsible for overcoming va resistance in TN 86. DNA sequence analysis revealed six amino acid differences between the two strains of TVMV within this domain. Inclusion of sequences for four of the TVMV-S VPg amino acids was sufficient to confer the resistance-overcoming phenotype to all corresponding transcripts. Coinoculation experiments suggested that the resistance of TN 86 to TVMV-WT was not due to the induction of a general host defense response. The results are compatible with the hypothesis that VPg must assume an appropriate configuration in order to interact with appropriate host components and facilitate systemic virus movement. PMID- 9356355 TI - The CM2 protein of influenza C virus is an oligomeric integral membrane glycoprotein structurally analogous to influenza A virus M2 and influenza B virus NB proteins. AB - We have undertaken a characterization of the CM2 protein of influenza C virus. The CM2 coding region of RNA segment 6 (nucleotides 731-1147) was cloned from two strains of influenza C virus and expressed using the vaccinia virus-bacteriophage T7 RNA polymerase (vac-T7) system. An antiserum raised to a C-terminal peptide in the CM2 open reading frame recognized the CM2 protein in influenza C virus infected cells and after vac-T7 expression of the CM2 open reading frame. CM2 is posttranslationally modified by addition of high-mannose carbohydrate chains (Mr approximately 18 kDa) and by further addition of polylactosaminoglycans (Mr approximately 21-35 kDa). The available data indicate that CM2 has a cleavable signal peptide at the N-terminus of the protein. Site-directed mutagenesis eliminated the single potential N-linked carbohydrate attachment site on CM2 and indicated that the protein has an NoutCin orientation in membranes. Nonreducing SDS-PAGE indicated that the protein was expressed as disulfide-linked dimers and tetramers. Cell surface biotinylation and indirect immunofluorescence showed the protein to be expressed at the cell surface. Elimination of the N-linked carbohydrate attachment site and addition of a C-terminal HA epitope tag did not adversely affect surface expression of CM2. The NoutCin membrane orientation of CM2, the size of the ectodomain and cytoplasmic tail of CM2, and its ability to form disulfide-linked oligomers are reminiscent of the structural properties of influenza A virus M2 and influenza B virus NB proteins. PMID- 9356377 TI - Hyperpolarization-activated inward current in neurons of the rat's dorsal nucleus of the lateral lemniscus in vitro. AB - Hyperpolarization-activated inward current in neurons of the rat's dorsal nucleus of the lateral lemniscus in vitro. J. Neurophysiol. 78: 2235-2245, 1997. The hyperpolarization-activated current (Ih) underlying inward rectification in neurons of the rat's dorsal nucleus of the lateral lemniscus (DNLL) was investigated using whole cell patch-clamp techniques. Patch recordings were made from DNLL neurons of young rats (21-30 days old) in 400 micro;m tissue slices. Under current clamp, injection of negative current produced a graded hyperpolarization of the cell membrane, often with a gradual sag in the membrane potential toward the resting value. The rate and magnitude of the sag depended on the amount of hyperpolarizing current. Larger current resulted in a larger and faster decay of the voltage. Under voltage clamp, hyperpolarizing voltage steps elicited a slowly activating inward current that was presumably responsible for the sag observed in the voltage response to a steady hyperpolarizing current recorded under current clamp. Activation of the inward current (Ih) was voltage and time dependent. The current just was seen at a membrane potential of -70 mV and was activated fully at -140 mV. The voltage value of half-maximal activation of Ih was -78.0 +/- 6.0 (SE) mV. The rate of Ih activation was best approximated by a single exponential function with a time constant that was voltage dependent, ranging from 276 +/- 27 ms at -100 mV to 186 +/- 11 ms at -140 mV. Reversal potential (Eh) of Ih current was more positive than the resting potential. Raising the extracellular potassium concentration shifted Eh to a more depolarized value, whereas lowering the extracellular sodium concentration shifted Eh in a more negative direction. Ih was sensitive to extracellular cesium but relatively insensitive to extracellular barium. The current amplitude near maximal-activation (about -140 mV) was reduced to 40% of control by 1 mM cesium but was reduced to only 71% of control by 2 mM barium. When the membrane potential was near the resting potential (about -60 mV), cesium had no effect on the membrane potential, current-evoked firing rate and input resistance but reduced the spontaneous firing. When the membrane potential was more negative than -70 mV, cesium hyperpolarized the cell, decreased current-evoked firing and increased the input resistance. Ih in DNLL neurons does not contribute to the normal resting potential but may enhance the extent of excitation, thereby making the DNLL a consistently powerful inhibitory source to upper levels of the auditory system. PMID- 9356379 TI - Parabrachial neural coding of taste stimuli in awake rats. AB - Parabrachial neural coding of taste stimuli in awake rats. J. Neurophysiol. 78: 2254-2268, 1997. In awake, behaving rats, the activity of 74 single neurons in the pontine parabrachial nucleus (PBN) was recorded in response to sapid stimulation by 15 chemicals. Of these, 44 taste cells were tested with all 15 stimuli. Based on their responsiveness to 4 standard stimuli, these neurons were categorized as follows: 23 NaCl-best, 15 sucrose-best, 5 citric acid-best, and 1 quinine HCl-best. Several forms of multivariate analyses indicated that the taste responses matched both the behavioral responses to and, less well, the chemical structure of, the sapid stimuli. A hierarchical cluster analysis of the neurons substantially confirmed the best-stimulus categorization, but separated the NaCl best cells into those that responded more to Na+-containing salts and those that responded more to Cl--containing salts. The cells that responded best to the Na+ moiety actually were somewhat more correlated with the sucrose-best cells than with those that responded to the Cl--containing stimuli. Citric acid-best neurons and the lone quinine-best unit formed a single cluster of neurons that responded well to acids, as well as to NH4Cl and, to a lesser extent, NaNO3. A factor analysis of the neuronal response profiles revealed that three factors accounted for 78.8% of the variance in the sample. Similar analyses of the stimuli suggested that PBN neurons respond to four or five sets of stimuli related by their chemical makeup or by human psychophysical reports. The capacity of rats to make these discriminations has been documented by other behavioral studies in which rodents generalize across sapid chemicals within each of 5 stimulus categories. Furthermore, a simulation analysis of the neural data replicated behavioral results that used amiloride, a Na+ channel blocker, in which rats generalized NaCl to non-Na+, Cl- salts. Thus, using a variety of analyses, in awake rats, the activity of PBN taste neurons tracks their behavioral responses to a variety of chemical stimuli. PMID- 9356378 TI - Contribution of outward currents to spike-frequency adaptation in hypoglossal motoneurons of the rat. AB - Contribution of outward currents to spike-frequency adaptation in hypoglossal motoneurons of the rat. J. Neurophysiol. 78: 2246-2253, 1997. Spike-frequency adaptation has been attributed to the actions of several different membrane currents. In this study, we assess the contributions of two of these currents: the net outward current generated by the electrogenic Na+-K+ pump and the outward current that flows through Ca2+-activated K+ channels. In recordings made from hypoglossal motoneurons in slices of rat brain stem, we found that bath application of a 4-20 microM ouabain solution produced a partial block of Na+-K+ pump activity as evidenced by a marked reduction in the postdischarge hyperpolarization that follows a period of sustained discharge. However, we observed no significant change in either the initial, early, or late phases of spike-frequency adaptation in the presence of ouabain. Adaptation also has been related to increases in the duration and magnitude of the medium-duration afterhyperpolarization (mAHP) mediated by Ca2+-activated K+ channels. When we replaced the 2 mM Ca2+ in the bathing solution with Mn2+, there was a significant decrease in the amplitude of the mAHP after a spike. The decrease in mAHP amplitude resulted in a decrease in the magnitude of the initial phase of spike frequency adaptation as has been reported previously by others. However, quite unexpectedly we also found that reducing the mAHP resulted in a dramatic increase in the magnitude of both the early and late phases of adaptation. These changes could be reversed by restoring the normal Ca2+ concentration in the bath. Our results with ouabain indicate that the Na+-K+ pump plays little, if any, role in the three phases of adaptation in rat hypoglossal motoneurons. Our results with Ca2+ channel blockade support the hypothesis that initial adaptation is, in part, controlled by conductances underlying the mAHP. However, our failure to eliminate initial adaptation completely by blocking Ca2+ channels suggests that other membrane mechanisms also contribute. Finally, the increase in both the early and late phases of adaptation in the presence of Mn2+ block of Ca2+ channels lends further support to the hypothesis that the initial and later (i.e., early and late) phases of spike-frequency adaptation are mediated by different cellular mechanisms. PMID- 9356381 TI - GABAA receptor-mediated Cl- currents in rat thalamic reticular and relay neurons. AB - GABAA receptor-mediated Cl- currents in rat thalamic reticular and relay neurons. J. Neurophysiol. 78: 2280-2286, 1997. Spontaneous and evoked inhibitory postsynaptic currents (sIPSCs and eIPSCs) and responses to exogenously applied gamma-aminobutyric acid (GABA), mediated by GABA type A (GABAA) receptors, were recorded in inhibitory neurons of nucleus reticularis thalami (nRt) and their target relay cells in ventrobasal (VB) nuclei by using patch clamp techniques in rat thalamic slices. The decay of sIPSCs in both nRt and VB neurons was best fitted with two exponential components. The decay time constants of sIPSCs in nRt neurons were much slower (tau1 = 38 ms; tau2 = 186 ms) than those previously reported in a variety of preparations and two to three times slower than those in VB neurons (tau1 = 17 ms; tau2 = 39 ms). GABAA receptor-mediated Cl- currents directly evoked by local GABA application also had a much slower decay time constant in nRt (225 ms) than in VB neurons (115 ms). Slow decay of GABA responses enhances the efficacy of recurrent intranuclear inhibition in nRt. The results suggest a functional diversity of GABAA receptors that may relate to the known heterogeneity of GABAA receptor subunits in these two thalamic nuclei. PMID- 9356380 TI - Properties of calcium spikes revealed during GABAA receptor antagonism in hippocampal CA1 neurons from guinea pigs. AB - Properties of calcium spikes revealed during GABAA receptor antagonism in hippocampal CA1 neurons from guinea pigs. J. Neurophysiol. 78: 2269-2279, 1997. Intracellular electrical responses and changes in intracellular calcium concentration ([Ca2+]i) in response to activation of synaptic inputs and to DC injections were recorded simultaneously from CA1 pyramidal neurons (n = 42) in guinea pig hippocampal slices. In the presence of the gamma-aminobutyric acid-A (GABAA) receptor antagonists, bicuculline (mu M) and picrotoxin (10 mu M, broad (>20 ms) all-or-none spikes were induced by activation of synaptic inputs (20 pulses, 30 Hz) and were accompanied by a simultaneous rapid and large rise in [Ca2+]i (34 of 34 cells). By contrast, direct depolarizing current (0.7 nA, 1 s) induced spikes having short duration, during which time the spike firing pattern was observed not to be significantly affected. When Na+ channels were blocked by QX-314 applied intracellularly through the recording microelectrode in the presence of GABAA receptor antagonists, broad spikes were frequently generated by activation of synaptic inputs (32 of 33 cells). These broad spikes were blocked by Cd2+ (200 mu M) or in Ca2+-free medium (6 of 6 cells) but were resistant to either tetrodotoxin (TTX; 1 micro M; 6 of 6 cells) or QX-314, whereas short duration spikes were blocked by both TTX and QX-314. Based on these findings we conclude that broad and short-duration spikes are Ca2+ and Na+ spikes, respectively. To investigate the properties of the Ca2+ spikes, antagonists of a voltage-operated Ca2+ channel were applied to the evoked responses. Nifedipine (30 mu M), a L-type Ca2+ channel blocker, suppressed the generation of Ca2+ spikes, whereas Ni2+ (100 mu M), the T- and R-type Ca2+ channel blocker, and omega-agatoxin-IVA (omega-Aga-IVA, 60 nM), a P-type Ca2+ channel blocker, had little effect on the generation of Ca2+ spikes. Nifedipine suppressed the rise in [Ca2+]i induced by synaptic inputs up to 26% of the control in the soma and 18 32% in the dendrites (n = 5), respectively, whereas Ni2+ suppressed the rise by 12-27% (n = 5) in both soma and dendrites. omega-Aga-IVA showed little effect (less than a 10% change; n = 7). These results suggest that the GABAA inhibitory system tonically suppresses dendritic Ca2+ spikes, and the L-type Ca2+ channel plays a major role in the generation of Ca2+ spikes and in Ca2+ influx. PMID- 9356382 TI - Metabotropic glutamate receptor enhancement of spontaneous IPSCs in neocortical interneurons. AB - Metabotropic glutamate receptor enhancement of spontaneous IPSCs in neocortical interneurons. J. Neurophysiol. 78: 2287-2295, 1997. Using neocortical layer I neurons as a model for GABAergic interneurons, we have studied gamma-aminobutyric acid-A (GABAA) receptor-mediated spontaneous inhibitory postsynaptic currents (IPSCs) and modulation by metabotropic glutamate receptors (mGluRs). In the presence of 0.5 mu M tetrodotoxin (TTX) and ionotropic glutamate receptor antagonists and under symmetrical Cl- conditions, the mean amplitude of miniature IPSCs (mIPSCs) was approximately 50 pA at a holding potential of -70 mV with individual events ranging from 10 to 400 pA. Averaged mIPSCs had a 10-90% rise time of approximately 0.6 ms. The decay was double exponential. The fast component had a time constant of approximately 4 ms and comprised approximately 40% of the total amplitude. The slow component had a time constant of approximately 22 ms. The frequency of spontaneous IPSCs (sIPSCs), recorded in the absence of TTX, was increased by bath application of the mGluR agonist 1S,3R-1 aminocyclopentane-1, 3-dicarboxylic acid (ACPD; 10-100 mu M) or the group I mGluR selective agonist quisqualic acid (Quis; 0.5-1 mu M). Under identical conditions, mIPSCs were not affected. The kinetics of sIPSCs and mIPSCs were not altered by ACPD or Quis. Quis (1 mu M) induced an inward current of approximately 70 pA at a holding potential of -70 mV, whereas ACPD (40-200 mu M) induced a smaller inward current. This current was linear over the voltage range -70 to +30 mV and reversed polarity near 0 mV. In current-clamp recordings, both Quis and ACPD induced a depolarization and action potential firing in layer I and deeper layer interneurons. We conclude that neocortical layer I neurons receive GABAA receptor mediated inhibitory synaptic inputs. Activation of mGluRs, possibly mGluR1 and/or mGluR5, causes an enhancement of inhibitory synaptic transmission by directly depolarizing corticalGABAergic interneurons through the opening of nonselective cation channels. PMID- 9356383 TI - Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia. AB - Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia. J. Neurophysiol. 78: 2296-2308, 1997. Allodynia is an unpleasant sequela of neural injury or neuropathy that is characterized by the inappropriate perception of light tactile stimuli as pain. This condition may be modeled experimentally in animals by the intrathecal (i.t.) administration of strychnine, a glycine receptor antagonist. Thus after i.t. strychnine, otherwise innocuous tactile stimuli evoke behavioral and autonomic responses that normally are elicited only by noxious stimuli. The current study was undertaken to determine how i.t. strychnine alters the spinal processing of somatosensory input by examining the responses of neurons in the ventroposterolateral thalamic nucleus. Extracellular, single-unit recordings were conducted in the lateral thalamus of 19 urethan-anaesthetized, male, Wistar rats (342 +/- 44 g; mean +/- SD). Receptive fields and responses to noxious and innocuous cutaneous stimuli were determined for 19 units (1 per animal) before and immediately after i.t. strychnine (40 microgram). Eighteen of the animals developed allodynia as evidenced by the ability of otherwise innocuous brush or air jet stimuli to evoke cardiovascular and/or motor reflexes. All (3) of the nociceptive-specific units became responsive to brush stimulation after i.t. strychnine, and one became sensitive to brushing over an expanded receptive field. Expansion of the receptive field, as determined by brush stimulation, also was exhibited by all of the low-threshold mechanoreceptive units (14) and wide dynamic range units (2) after i.t. strychnine. The use of air jet stimuli at fixed cutaneous sites also provided evidence of receptive field expansion, because significant unit responses to air jet developed at 13 cutaneous sites (on 7 animals) where an identical stimulus was ineffective in evoking a unit response before i.t. strychnine. However, the magnitude of the unit response to cutaneous air jet stimulation was not changed at sites that already had been sensitive to this stimulus before i.t. strychnine. The onset of allodynia corresponded with the onset of the altered unit responses (i.e., lowered threshold/receptive field expansion) for the majority of animals (9), but the altered unit response either terminated concurrently with symptoms of allodynia (6) or, more frequently, outlasted the symptoms of allodynia (10) as the effects of strychnine declined. The present results demonstrate that the direct, receptor-mediated actions of strychnine on the spinal processing of sensory information are reflected by changes in the receptive fields and response properties of nociceptive and nonnociceptive thalamic neurons. These changes are consistent with the involvement of thalamocortical mechanisms in the expression of strychnine-induced allodynia and, moreover, suggest that i.t. strychnine also produces changes in innocuous tactile sensation. PMID- 9356384 TI - Three kinetically distinct Ca2+-independent depolarization-activated K+ currents in callosal-projecting rat visual cortical neurons. AB - Three kinetically distinct Ca2+-independent depolarization-activated K+ currents in callosal-projecting rat visual cortical neurons. J. Neurophysiol. 78: 2309 2320, 1997. Whole cell, Ca2+-independent, depolarization-activated K+ currents were characterized in identified callosal-projecting (CP) neurons isolated from postnatal day 7-16 rat primary visual cortex. CP neurons were identified in vitro after in vivo retrograde labeling with fluorescently tagged latex microbeads. During brief (160-ms) depolarizing voltage steps to potentials between -50 and +60 mV, outward K+ currents in these cells activate rapidly and inactivate to varying degrees. Three distinct K+ currents were separated based on differential sensitivity to 4-aminopyridine (4-AP); these are referred to here as IA, ID, and IK, because their properties are similar (but not identical) K+ currents termed IA, ID, and IK in other cells. The current sensitive to high (>/=100 mu M) concentrations of 4-AP (IA) activates and inactivates rapidly; the current blocked completely by low (100 mu M), response amplitudes were suppressed below control levels. The enhancement of GABAA receptor activity on skate bipolar cells showed little voltage dependence, suggesting that zinc is acting on the extracellular domain of the GABAA receptor. In the presence of 10 mu M zinc, the dose-response curve for 4,5,6, 7-tetrahydroisoxazolo[5,4-c]pyridin 3-ol (THIP; a GABAA agonist that suppresses GABAC-activated currents) was shifted to the left of the curve obtained in the absence of zinc, but without a significant change in the response maximum. This finding indicates that the enhancing effect of zinc is due primarily to its ability to increase the sensitivity of the GABAA receptor. The novel enhancement of neuronal GABAA receptor activity by zinc, observed previously in the GABAA-mediated responses of skate Muller (glial) cells, may reflect the presence of a unique subtype of GABAA receptor on the bipolar and Muller cells of the skate retina. PMID- 9356393 TI - Reaching movements with similar hand paths but different arm orientations. II. Activity of individual cells in dorsal premotor cortex and parietal area 5. AB - Reaching movements with similar hand paths but different arm orientations. II. Activity of individual cells in dorsal premotor cortex and parietal area 5. J. Neurophysiol. 78: 2413-2426, 1997. Neuronal activity in primary motor cortex (MI) is altered when monkeys make reaching movements along similar handpaths at shoulder level with two different arm orientations, either in the natural orientation with the elbow positioned below the level of the shoulder and hand or in an abducted orientation with the elbow abducted nearly to shoulder level. The present study examines to what degree two other cortical areas, the dorsal premotor (PMd) and parietal area 5, also show modulation of cell activity related to arm geometry during reaching. The activity of most (89%) of the 207 cells in PMd recorded while monkeys made reaching movements showed a statistically significant change in activity between orientations [analysis of variation (ANOVA), P < 0.01]. A common effect of arm orientation on cell activity was a change in the overall level of discharge either before, during, and/or after movement (67%, ANOVA, task main effect, P < 0.01). Many cells (76%) showed a statistical change in their response to movement direction (ANOVA, task x direction interaction term, P < 0.01), including changes in dynamic range and changes in the preferred direction of cells that were directionally tuned in both arm orientations. Overall, these effects were similar qualitatively but not as strong quantitatively as those observed in MI. A sample of cells was recorded in area 5 of one monkey. Most (95%) of the 79 area 5 cells showed a change in activity when reaching movements were performed using different arm orientations (ANOVA, P < 0.01). As in PMd and MI, many area 5 cells (56, 71%) showed changes in their tonic discharge before, during, and/or after movement, and 70 cells (89%) showed changes in their response to movement direction (ANOVA, task x direction interaction term, P < 0.01). The observed changes in neuronal activity related to posture and movement in MI, PMd and area 5 demonstrate that single cell activity in these cortical areas is not simply related to the spatial attributes of hand trajectory but is also strongly influenced by attributes of movement related to arm geometry. PMID- 9356394 TI - Electrophysiological and morphological characteristics of neurons in perinuclear zone of supraoptic nucleus. AB - Electrophysiological and morphological characteristics of neurons in perinuclear zone of supraoptic nucleus. J. Neurophysiol. 78: 2427-2437, 1997. Neurons in the perinuclear zone (PZ) of the supraoptic nucleus (SON) are thought to serve as interneurons and may mediate changes in neurohypophysial hormone release in response to physiological changes in blood pressure. However, the morphology and electrophysiological characteristics of PZ neurons are unknown. In the present study, PZ neurons from male and female rats were recorded intracellularly to determine some membrane properties, then filled with biocytin or biotinamide for morphological analysis. In general, PZ neurons had faster spikes than magnocellular SON neurons, and the great majority were characterized by a subthreshold depolarizing hump when depolarized from a hyperpolarized (less than 80 mV) membrane potential. In most neurons, this hump was similar to low threshold spikes described in other CNS regions. Near-threshold, fast action potentials were clustered near the onset of these depolarizations. Conspicuously absent in all PZ neurons was the strong transient and subthreshold outward rectification characteristic of vasopressin and oxytocin neurons of the SON. These results suggest that PZ neurons are electrophysiologically distinct from neurosecretory neurons of the SON. No differences were found between male and female rats in any of the basic properties examined, including input resistance, membrane time constant, spike height, spike width, spike threshold, and the size of the spike afterhyperpolarization. Morphologically, PZ neurons were diverse but were divided into spiny and aspiny groups. Three spiny neurons and one aspiny neuron contributed an axonal projection to the SON characterized by varicosities suggestive of terminals. In the case of the three spiny neurons, the SON projection was clearly a minor collateral projection. The axon arborized in the PZ, but one or more branches were cut at the edge of the explant, indicating a longer projection. In the remaining neurons, no axonal projection to the SON was detected and several had axons leaving the explant. Some portion of the dendritic tree penetrated the SON in several neurons. The morphology of PZ neurons was thus heterogeneous and suggests that, for some cells at least, the projection to the SON may be a minor collateral component of a much wider axonal projection. PMID- 9356395 TI - First-spike timing of auditory-nerve fibers and comparison with auditory cortex. AB - First-spike timing of auditory-nerve fibers and comparison with auditory cortex. J. Neurophysiol. 78: 2438-2454, 1997. The timing of the first spike of cat auditory-nerve (AN) fibers in response to onsets of characteristic frequency (CF) tone bursts was studied and compared with that of neurons in primary auditory cortex (AI), reported previously. Tones were shaped with cosine-squared rise functions, and rise time and sound pressure level were parametrically varied. Although measurement of first-spike latency of AN fibers was somewhat compromised by effects of spontaneous activity, latency was an invariant and inverse function of the maximum acceleration of peak pressure (i.e., a feature of the 2nd derivative of the stimulus envelope), as previously found in AI, rather than of tone level or rise time. Latency-acceleration functions of all AN fibers were of very similar shape, similar to that observed in AI. As in AI, latency acceleration functions of different fibers were displaced along the latency axis, reflecting differences in minimum latency, and along the acceleration axis, reflecting differences in sensitivity to acceleration [neuronal transient sensitivity (S)]. S estimates increased with spontaneous rate (SR), but values of high-SR fibers exceeded those in AI. This suggests that S estimates are biased by SR per se, and that unbiased true S values would be less tightly correlated with response properties covarying with SR, such as firing threshold. S estimates varied with CF in a fashion similar to the cat's audiogram and, for low- and medium-SR fibers, matched those for AI neurons. Minimum latency decreased with increasing SR and CF. As in AI, the standard deviation of first-spike timing (SD) in AN was also an inverse function of maximum acceleration of peak pressure. The characteristics of the increase of SD with latency in a given AN fiber/AI neuron and across AN fibers/AI neurons revealed that the precision of first-spike timing to some stimuli can actually be higher in AI than in AN. The data suggest that the basic characteristics of the latency-acceleration functions of transient onset responses seen in cortex are generated at inner hair cell-AN fiber synapses. Implications for signal processing in the auditory system and for first spike generation and adaptation in AN are discussed. PMID- 9356396 TI - Electrical properties of a cockroach motor neuron soma depend on different characteristics of individual Ca components. AB - Electrical properties of a cockroach motor neuron soma depend on different characteristics of individual Ca components. J. Neurophysiol. 78: 2455-2466, 1997. The "fast" coxal depressor motor neuron (Df) of the cockroach is among the most extensively studied of insect neurons. It has been shown that the cell body of this neuron can exhibit active electrical properties, which may change over time or with chemical modulation. To further understand these electrical events and their modulation, inward currents in Df have been characterized under conditions in which outward currents have been suppressed. The inward current activated at potentials positive to -60 mV and peaked between -10 and 0 mV when measured in barium saline and between 0 and +10 mV when measured in calcium saline. The inward current was insensitive to Ni2+ (100 mu M) but reduced by verapamil (50 mu M) and abolished by Cd2+ (1 mM). Two components of ICa were identified by their sensitivity to either 50 mu M nifedipine or micromolar Cd2+. The nifedipine-sensitive component activated positive to -60 mV and peaked between -10 and 0 mV, whereas the Cd2+-sensitive component activated positive to 40 mV and peaked between +10 and +20 mV. Immediately after dissection, depolarization of Df evoked plateau potentials, whereas 1-4 h after dissection, depolarization evoked action potentials. The plateau potentials were insensitive to 100 mu M Cd2+ but blocked by 50 mu M nifedipine, whereas the spikes required a combination of nifedipine (50 mu M) and Cd2+ (100 mu M) for complete suppression, indicating that only one component of ICa contributes to the plateau potential, whereas both components contribute to action potentials. Currents measured in calcium saline decayed faster than currents measured in barium saline. The inactivation characteristics were investigated with the use of double-pulse voltage-clamp experiments. ICa showed a greater degree of inactivation and slower recovery from inactivation than did IBa. Current decay and the extent of inactivation were reduced after injection of the calcium-chelator 1,2-bis(2 aminophenoxy)ethane-N,N, N',N'-tetraacetic acid (BAPTA). This suggests that the calcium current of this neuron displays calcium-dependent inactivation. An additional mechanism, most probably voltage-dependent inactivation, also occurs because IBa, even in neurons injected with BAPTA, displayed some inactivation. The inactivation characteristics may be important in determining activity displayed by Df. Indirect evidence suggests that intracellular calcium is high immediately after dissection. At this time, the calcium current may therefore be reduced due to calcium-dependent inactivation. This may, at least partly, explain why the cell does not spike shortly after dissection. PMID- 9356398 TI - Interaction between tetanus long-term potentiation and hypoxia-induced potentiation in the rat hippocampus. AB - Interaction between tetanus long-term potentiation and hypoxia-induced potentiation in the rat hippocampus. J. Neurophysiol. 78: 2475-2482, 1997. The interaction between tetanus-induced long-term potentiation (LTP) and hypoxia induced potentiation was investigated by performing extracellular recordings in the CA1 region of rat hippocampus using a two-pathway design. Hippocampal slices were placed in an interface chamber containing artificial cerebrospinal fluid (ACSF) solution with high magnesium concentration. Hypoxia was induced by replacing the 5% CO2-95% O2 gas mixture with 5% CO2-95% N2 for 2 min. Tetanus-LTP was induced with 1-s, 100-Hz current pulses. Significant hypoxia-induced potentiation of the slope of the dendritic excitatory postsynaptic potential (EPSP) was found in ACSF containing 2 mM of magnesium 2, 27 +/- 10% (mean +/- SE; n = 16; P < 0.01) with no change in the mean amplitude of the presynaptic volley. All experiments in which a stable control baseline was obtained were used for data analysis. The data show that short episodes (2 min) of hypoxia can induce LTP of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-mediated synaptic transmission. The present study demonstrated that after tetanus-LTP, 33 +/- 3% (n = 10; P < 0.01), hypoxia further potentiated the field EPSP slopes by a mean value of 16 +/- 5% (n = 10; P < 0.05). Moreover, using a two-pathway design, we showed that hypoxia produced similar potentiation in both the control [19 +/- 5%; n = 10; P < 0.01) and tetanus-induced LTP pathway, and the total potentiation produced by a combination of tetanus then hypoxia, 63 +/- 13% (n = 10; p < 0.01), was significantly larger (P < 0.01) than hypoxia alone. These data suggest that hypoxia-induced potentiation is additive with tetanus-LTP. Occlusion experiments were performed to verify whether the mechanisms responsible for hypoxia-induced potentiation are independent of preexisting synaptic levels induced by high frequency stimulation. Hypoxia produced significant potentiation (23 +/- 7%; n = 7; P < 0.05) after successful occlusion of the LTP pathway. Therefore, because the magnitude of hypoxia-induced potentiation is both independent of preexisting synaptic levels and also additive, synaptic specificity associated with LTP is preserved. The magnitude of tetanus-LTP induced 20 min after hypoxia (15 +/- 4%; n = 10) was significantly smaller (P < 0.01) relative to LTP after normoxic conditions (33 +/- 3%; n = 10). Additionally, hypoxia blocked the transient, robust potentiation occurring during the early phase of LTP induction. This study suggests that although hypoxia modifies neuronal processing by general excitation, synaptic specificity associated with tetanus-LTP still is preserved. However, hypoxia can disrupt neuronal processing by inhibiting new modification of synaptic transmission. PMID- 9356397 TI - Ca2+ current in rabbit carotid body glomus cells is conducted by multiple types of high-voltage-activated Ca2+ channels. AB - Ca2+ current in rabbit carotid body glomus cells is conducted by multiple types of high-voltage-activated Ca2+ channels. J. Neurophysiol. 78: 2467-2474, 1997. Carotid bodies are sensory organs that detect changes in arterial oxygen. Glomus cells are presumed to be the initial sites for sensory transduction, and Ca2+ dependent neurotransmitter release from glomus cells is believed to be an obligatory step in this response. Some information exists on the Ca2+ channels in rat glomus cells. However, relatively little is known about the types of Ca2+ channels present in rabbit glomus cells, the species in which most of the neurotransmitter release studies have been performed. Therefore we tested the effect of specific Ca2+ channel blockers on current recorded from freshly dissociated, adult rabbit carotid body glomus cells using the whole cell configuration of the patch-clamp technique. Macroscopic Ba2+ current elicited from a holding potential of -80 mV activated at a Vm of approximately -30 mV, peaked between 0 and +10 mV and did not inactivate during 25-ms steps to positive test potentials. Prolonged ( approximately 2 min) depolarized holding potentials inactivated the current with a V1/2 of -47 mV. There was no evidence for T-type channels. On steps to 0 mV, 6 mM Co2+ decreased peak inward current by 97 +/- 1% (mean +/- SE). Nisoldipine (2 mu M), 1 mu M omega-conotoxin GVIA, and 100 nM omega-agatoxin IVa each blocked a portion of the macroscopic Ca2+ current (30 +/- 5, 33 +/- 5, and 19 +/- 3% after rundown correction, respectively). Simultaneous application of these blockers revealed a resistant current that was not affected by 1 mu M omega-conotoxin MVIIC. This resistant current constituted 27 +/- 5% of the total macroscopic Ca2+ current. Each blocker had an effect in every cell so tested. However, the relative proportion of current blocked varied from cell to cell. These results suggest that L, N, P, and resistant channel types each conduct a significant proportion of the macroscopic Ca2+ current in rabbit glomus cells. Hypoxia-induced neurotransmitter release from glomus cells may involve one or more of these channels. PMID- 9356399 TI - Calcium-dependent plateau potentials in rostral ambiguus neurons in the newborn mouse brain stem in vitro. AB - Calcium-dependent plateau potentials in rostral ambiguus neurons in the newborn mouse brain stem in vitro. J. Neurophysiol. 78: 2483-2492, 1997. The nucleus ambiguus contains vagal and glossopharyngeal motoneurons and preganglionic neurons involved in respiration, swallowing, vocalization, and control of heart beat. Here we show that the rostral compact formation's ambiguus neurons, which control the esophageal phase of swallowing, display calcium-dependent plateau potentials in response to tetanic orthodromic stimulation or current injection. Whole cell recordings were made from visualized neurons in the rostral nucleus ambiguus using a slice preparation from the newborn mouse. Biocytin-labeling revealed dendritic trees with pronounced rostrocaudal orientations confined to the nucleus ambiguus, a morphological profile matching that of vagal motoneurons projecting to the esophagus. Single-stimulus orthodromic activation, using an electrode placed in the dorsomedial slice near the nucleus tractus solitarius, evoked single excitatory postsynaptic potentials (EPSPs) or short trains of EPSPs (500 ms to 1 s). However, tetanic stimulation (5 pulses, 10 Hz) induced voltage dependent afterdepolarizations or long-lasting plateau potentials (>1 min) with a constant firing pattern. Depolarizing or hyperpolarizing current pulses elicited voltage-dependent afterdepolarizations or plateau potentials lasting a few seconds to several minutes. Constant spike activity accompanied the long-lasting plateau potentials, which ended spontaneously or could be terminated by weak hyperpolarizing current pulses. Current-induced afterdepolarizations and plateau potentials were dependent on extracellular and intracellular Ca2+, as they were blocked completely by extracellular Co2+, Cd2+, or intracellular bis-(o aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA). Orthodromically induced afterdepolarizations and plateau potentials were blocked by intracellular BAPTA. Afterdepolarizations and plateau potentials were completely blocked by substitution of extracellular Na+ with choline. Afterdepolarizations persisted in tetrodotoxin. We conclude that rostral ambiguus neurons have a Ca2+-activated inward current carried by Na+. Synaptic activation of this conductance may generate prolonged spike activity in these neurons during the esophageal phase of swallowing. PMID- 9356400 TI - Hippocampal interneurons are excited via serotonin-gated ion channels. AB - Hippocampal interneurons are excited via serotonin-gated ion channels. J. Neurophysiol. 78: 2493-2502, 1997. Serotonergic neurons of the median raphe nucleus heavily innervate hippocampal GABAergic interneurons located in stratum radiatum of area CA1, suggesting that this strong subcortical projection may modulate interneuron excitability. Using whole cell patch-clamp recording from interneurons in brain slices, we tested the effects of serotonin (5-HT) on the physiological properties of these interneurons. Serotonin produces a rapid inward current that persists when synaptic transmission is blocked by tetrodotoxin and cobalt, and is unaffected by ionotropic glutamate and gamma-aminobutyric acid (GABA) receptor antagonists. The 5-HT-induced current was independent of G protein activation. Pharmacological evidence indicates that 5-HT directly excites these interneurons through activation of 5-HT3 receptors. At membrane potentials negative to -55 mV, the current-voltage (I-V) relationship of the 5-HT current displays a region of negative slope conductance. Therefore the response of interneurons to 5-HT strongly depends on membrane potential and increases greatly as cells are depolarized. Removal of extracellular calcium, but not magnesium, increases the amplitude of 5-HT-induced currents and removes the region of negative slope conductance, thereby linearizing the I-V relationship. The axons of 5-HT-responsive interneurons ramify widely within CA1; some of these interneurons also project to and arborize extensively in the dentate gyrus. The organization of these inhibitory connections strongly suggests that these cells regulate excitability of both CA1 pyramidal cells and dentate granule cells. As our results indicate that 5-HT may mediate fast excitatory synaptic transmission onto these interneurons, serotonergic inputs can simultaneously modulate the output of both hippocampus and dentate gyrus. PMID- 9356401 TI - Monkey cutaneous SAI and RA responses to raised and depressed scanned patterns: effects of width, height, orientation, and a raised surround. AB - Monkey cutaneous SAI and RA responses to raised and depressed scanned patterns: effects of width, height, orientation, and a raised surround. J. Neurophysiol. 78: 2503-2517, 1997. The aim of this study was to examine the slowly adapting type I (SAI) and rapidly adapting (RA) primary afferent representation of raised and depressed surface features. Isolated, raised, and depressed squares and small raised squares with a circular surround were scanned across the receptive fields of SAI and RA mechanoreceptive afferents innervating the distal fingerpads of the rhesus monkey. Pattern height ranged from -620 to +620 micron and width ranged from 0.2 to 7.0 mm. The surround radii ranged from 3.0 to 7.0 mm. Previous combined psychophysical and neurophysiological studies have provided evidence that SAI afferent responses are responsible for the perception of spatial form and texture and that RA afferents are responsible for the detection of stimuli that produce minute skin motion (flutter, slip, microgeometric surface features). Our results strengthen these hypotheses. Response properties shared by both SAI and RA afferent types were that both responded only to the edges of the larger raised and depressed patterns, both responded to falling edges half as vigorously as to rising edges, both responded to rising and falling edges with impulse rates that were proportional to the sine of the angle between the edge and the scanning direction, and both had suppressed responses to a small raised surface feature when a raised surround was closer than 6 mm. Response differences consistent with the hypothesis that SAI afferents are specialized for the representation of form were that SAI responses were confined to areas around the features that evoked them in areas that were 40-50% smaller than the comparable RA response areas, SAI responses were more than four times more sensitive to stimulus height than were RA afferents over the range from 280 to 620 micron, and SAI (but not RA) afferents responded 20-50% more vigorously to corners than to edges. Response differences consistent with the hypothesis that RA afferents are specialized for the detection of minute surfaces features were that only RA afferents responded to very small surface depressions, depressed squares 0.8 mm wide, that were detectable by palpation. Mechanisms underlying the many differences in SAI and RA response properties are discussed. PMID- 9356402 TI - Quantal organization of reflex and conditioned eyelid responses. AB - Quantal organization of reflex and conditioned eyelid responses. J. Neurophysiol. 78: 2518-2530, 1997. Upper lid movements and the electromyographic activity of the orbicularis oculi muscle were recorded in behaving cats during spontaneous and experimentally evoked reflex blinks, and conditioned eyelid responses. Reflex blinks evoked by the presentation of air puffs, flashes, or tones consisted of a fast downward lid movement followed by late, small downward waves, recurring at approximately 50-ms intervals. The latency, maximum amplitude, peak velocity, and number of late waves depended on the modality, intensity, and duration of the evoking stimulus. The power spectra of acceleration records indicated a dominant frequency of approximately 20 Hz for air puff-evoked blinks. Flashes and tones usually evoked small and easily fatigable reflex responses of lower dominant frequencies (14-17 and 9-11 Hz, respectively). A basic approximately 20-Hz oscillation was also noticed during lid fixation, and ramplike lid displacements evoked by optokinetic stimuli. Five classical conditioning paradigms were used to analyze the frequency-domain properties of conditioned eyelid responses. These learned lid movements differed in latency, maximum amplitude, and profile smoothness depending on the modality (air puff, tone), intensity (weak, strong), and presentation site (ipsi-, contralateral to the unconditioned stimulus) of the conditioned stimulus. It was found that the characteristic ramplike profile of a conditioned response was not smooth, but appeared to be formed by a succession of small waves at a dominant frequency of approximately 20 Hz. The amplitude (and number) of the constituting waves depended on the characteristics of the conditioned stimulus and on the time interval until unconditioned stimulus presentation. Thus conditioned responses seemed to be formed from lid displacements of 2-6 degrees in amplitude and approximately 50 ms in duration, which increased in number throughout conditioning sessions, until a complete (i.e., lid closing) conditioned response was reached. It is suggested that a approximately 20-Hz oscillator underlies the generation of reflex and conditioned eyelid responses. The oscillator is susceptible to being neurally modulated to modify the velocity of a given quantum of movement, and the total duration of the lid response. Learned eyelid movements are probably the result of a successively longer release of the oscillator as a function of the temporal-spatial needs of the motor response. PMID- 9356404 TI - Regulation of the NMDA component of EPSPs by different components of postsynaptic GABAergic inhibition: computer simulation analysis in piriform cortex. AB - Regulation of the NMDA component of EPSPs by different components of postsynaptic GABAergic inhibition: computer simulation analysis in piriform cortex. J. Neurophysiol. 78: 2546-2559, 1997. Physiological analysis in the companion paper demonstrated that gamma-aminobutyric acid-A (GABAA)-mediated inhibition in piriform cortex is generated by circuits that are largely independent in apical dendritic and somatic regions of pyramidal cells and that GABAA-mediated inhibitory postsynaptic currents (IPSCs) in distal dendrites have a slower time course than those in the somatic region. This study used modeling methods to explore these characteristics of GABAA-mediated inhibition with respect to regulation of the N-methyl--aspartate (NMDA) component of excitatory postsynaptic potentials. Such regulation is relevant to understanding NMDA-dependent long-term potentiation (LTP) and the integration of repetitive synaptic inputs that can activate the NMDA component as well as pathological processes that can be activated by overexpression of the NMDA component. A working hypothesis was that the independence and differing properties of IPSCs in apical dendritic and somatic regions provide a means whereby the NMDA component and other dendritic processes can be controlled by way of GABAergic tone without substantially altering system excitability. The analysis was performed on a branched compartmental model of a pyramidal cell in piriform cortex constructed with physiological and anatomic data derived by whole cell patch recording. Simulations with the model revealed that NMDA expression is more effectively blocked by the slow GABAA component than the fast. Because the slow component is present in greater proportion in apical dendritic than somatic regions, this characteristic would increase the capacity of dendritic IPSCs to regulate NMDA mediated processes. The simulations further revealed that somatic-region GABAergic inhibition can regulate the generation of action potentials with little effect on the NMDA component generated by afferent fibers in apical dendrites. As a result, if expression of the NMDA component or other dendritic processes were enabled by selective block of dendritic inhibition, for example, by centrifugal fiber systems that may regulate learning and memory, the somatic-region IPSC could preserve system stability through feedback regulation of firing without counteracting the effect of the dendritic-region block. Simulations with paired inputs revealed that the dendritic GABAA-mediated IPSC can regulate the extent to which a strong excitatory input facilitates the NMDA component of a concurrent weak input, providing a possible mechanism for control of "associative LTP" that has been demonstrated in this system. Postsynaptic GABAB-mediated inhibition had less effect on the NMDA component than either the fast or slow GABAA components. Depolarization from a concomitant alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) component also was found to have comparatively little effect on current through the NMDA channel because of its brief time course. PMID- 9356403 TI - GABAA-mediated IPSCs in piriform cortex have fast and slow components with different properties and locations on pyramidal cells. AB - GABAA-mediated IPSCs in piriform cortex have fast and slow components with different properties and locations on pyramidal cells. J. Neurophysiol. 78: 2531 2545, 1997. A recent study in piriform (olfactory) cortex provided evidence that, as in hippocampus and neocortex, gamma-aminobutyric acid-A (GABAA)-mediated inhibition is generated in dendrites of pyramidal cells, not just in the somatic region as previously believed. This study examines selected properties of GABAA inhibitory postsynaptic currents (IPSCs) in dendritic and somatic regions that could provide insight into their functional roles. Pharmacologically isolated GABAA-mediated IPSCs were studied by whole cell patch recording in slices. To compare properties of IPSCs in distal dendritic and somatic regions, local stimulation was carried out with tungsten microelectrodes, and spatially restricted blockade of GABAA-mediated inhibition was achieved by pressure ejection of bicuculline from micropipettes. The results revealed that largely independent circuits generate GABAA inhibition in distal apical dendritic and somatic regions. With such independence, a selective decrease in dendritic-region inhibition could enhance integrative or plastic processes in dendrites while allowing feedback inhibition in the somatic region to restrain system excitability. This could allow modulatory fiber systems from the basal forebrain or brain stem, for example, to change the functional state of the cortex by altering the excitability of interneurons that mediate dendritic inhibition without increasing the propensity for regenerative bursting in this highly epileptogenic system. As in hippocampus, GABAA-mediated IPSCs were found to have fast and slow components with time constants of decay on the order of 10 and 40 ms, respectively, at 29 degrees C. Modeling analysis supported physiological evidence that the slow time constant represents a true IPSC component rather than an artifactual slowing of the fast component from voltage clamp of a dendritic current. The results indicated that, whereas both dendritic and somatic-region IPSCs have both fast and slow GABAA components, there is a greater proportion of the slow component in dendrites. In a companion paper, the hypothesis is explored that the resulting slower time course of the dendritic IPSC increases its capacity to regulate the N-methyl--aspartate component of EPSPs. Finally, evidence is presented that the slow GABAA-mediated IPSC component is regulated by presynaptic GABAB inhibition whereas the fast is not. Based on the requirement for presynaptic GABAB-mediated block of inhibition for expression of long-term potentiation, this finding is consistent with participation of the slow GABAA component in regulation of synaptic plasticity. The lack of susceptibility of the fast GABAA component to the long-lasting, activity-induced suppression mediated by presynaptic GABAB receptors is consistent with a protective role for this process in preventing seizure activity. PMID- 9356405 TI - Receptive properties of embryonic chick sensory neurons innervating skin. AB - Receptive properties of embryonic chick sensory neurons innervating skin. J. Neurophysiol. 78: 2560-2568, 1997. We describe a new in vitro skin-nerve preparation from chick embryos that allows detailed study of the functional properties of developing sensory neurons innervating skin. Functionally single sensory afferents were isolated by recording from their axons in microdissected filaments of the cutaneous femoralis medialis nerve, which innervates skin of the thigh. A total of 157 single neurons were characterized from embryos [embryonic days 17-21 (E17-E21), n = 115] and hatchlings up to 3 wk old (n = 42). Neurons were initially classified on the basis of their conduction velocity; those conducting below 1.0 m/s were being classified as C fibers and faster conducting fibers as A fibers. The proportions of A and C fibers encountered in embryonic and hatchling preparations were not very different, indicating that myelination and axon growth proceeds quite slowly over the period studied. Afferent fibers that could subserve nociceptive and nonnociceptive functions were identified in the time period studied. Subpopulations of low-threshold myelinated afferent units exhibited rapidly or slowly adapting discharges to constant force stimuli and could have tactile functions. Many afferent fibers responded to noxious heat and were excited and sensitized by exposure to inflammatory mediators, suggesting that they are nociceptors. The behavior of these units changed in several respects over the period studied. The discharge of C fibers to noxious heat increased with age as did their mechanical thresholds. A substantial population of heat-responsive neurons (34% of the A fibers) present in embryos were not encountered in hatchling chicks. This indicates that substantial changes in the physiological response properties of sensory afferents occur after hatching. We conclude that this new preparation can be used for quantitative assessment of the receptive properties of developing sensory neurons and has considerable potential for the investigation of factors, such as neurotrophins, that specify and influence the functional phenotype of sensory neurons during embryonic development in vivo. PMID- 9356406 TI - Conditions for the induction of long-term potentiation and long-term depression by conjunctive pairing in the dentate gyrus in vitro. AB - Conditions for the induction of long-term potentiation and long-term depression by conjunctive pairing in the dentate gyrus in vitro. J. Neurophysiol. 78: 2569 2573, 1997. The conditions under which long-term potentiation (LTP) and long-term depression (LTD) of excitatory postsynaptic currents were induced by the conjunctive pairing-type protocol of afferent stimulation and postsynaptic depolarization were studied in the medial perforant pathway-granule cell synapse of the dentate gyrus in vitro. The conjunctive pairing of 1-Hz afferent stimulation and steady state postsynaptic depolarization to 0 mV did not induce LTP or LTD. Inhibition of LTD induction with a phosphatase inhibitor or ruthenium red resulted in induction of LTP after the conjunctive pairing. Such LTP induction was N-methyl--aspartate dependent. Conversely, inhibition of LTP induction with a kinase inhibitor resulted in LTD induction after the conjunctive pairing. Thus the failure to induce LTP or LTD with the pairing protocol involving depolarization to 0 mV membrane potential was due to simultaneous activation of intracellular processes that generate the induction of LTP and LTD. Increasing the frequency of afferent stimulation to 200 Hz, even for just eight stimuli, resulted in LTP induction. The studies show that two factors govern the induction of LTP/LTD, membrane potential and frequency of afferent stimulation, with either increased depolarization or increased afferent stimulation favoring LTP induction. PMID- 9356407 TI - LTP induction dependent on activation of Ni2+-sensitive voltage-gated calcium channels, but not NMDA receptors, in the rat dentate gyrus in vitro. AB - LTP induction dependent on activation of Ni2+-sensitive voltage-gated calcium channels, but not NMDA receptors, in the rat dentate gyrus in vitro. J. Neurophysiol. 78: 2574-2581, 1997. A N-methyl--aspartate receptor (NMDAR) independent long-term potentiation (LTP) has been investigated in the dentate gyrus of the hippocampus in vitro in the presence of the NMDAR antagonist, -2 amino-phosphonopentanoate (50-100 mu M), at a concentration that completely blocked NMDAR-mediated excitatory postsynaptic currents (EPSCs). LTP of patch clamped EPSCs was induced by pairing low-frequency evoked EPSCs (1 Hz) with depolarizing voltage pulses designed to predominately open low-voltage-activated (LVA) Ca2+ channels. Voltage pulses alone induced only a short-term potentiation. The LTP was blocked by intracellular application of the rapid Ca2+ chelator bis (o-aminophenoxy)-N,N,N',N'-tetraacetic acid, demonstrating that a rise in intracellular Ca2+ is required for the NMDAR-independent LTP induction. The NMDAR independent LTP induction also was blocked by Ni2+ at a low extracellular concentration (50 mu M), which is known to strongly block LVA Ca2+ channels. However, Ni2+ did not inhibit the NMDAR-dependent LTP induced by high-frequency stimulation (HFS). The NMDAR-independent LTP induction was not blocked by high concentrations of the L-type Ca2+ channel blocker nifedipine (10 mu M). The NMDAR independent LTP was inhibited by the metabotropic glutamate receptor ligand (+) alpha-methyl-4-carboxyphenylglycine. These experiments demonstrate the presence of a NMDAR-independent LTP induced by Ca2+ influx via Ni2+-sensitive, nifedipine insensitive voltage-gated Ca2+ channels, probably LVA Ca2+ channels. Induction of the NMDAR-independent LTP was inhibited by prior induction of HFS-induced NMDAR dependent LTP, demonstrating that although the NMDAR-dependent and NMDAR independent LTP use a different Ca2+ channel for Ca2+ influx, they share a common intracellular pathway. PMID- 9356408 TI - Ionic mechanisms of spontaneous GABAergic events in rat hippocampal slices exposed to 4-aminopyridine. AB - Ionic mechanisms of spontaneous GABAergic events in rat hippocampal slices exposed to 4-aminopyridine. J. Neurophysiol. 78: 2582-2591, 1997. Ion-selective (H+ and K+) microelectrode techniques as well as conventional extra- and intracellular recordings were used to study the ionic mechanisms of propagating spontaneous GABAergic events (SGEs) in rat hippocampal slices exposed to 4 aminopyridine (4-AP, 50-100 mu M). All experiments were made in the presence of antagonists of ionotropic glutamate receptors [10 mu M 6-nitro-7 sulphamoylbenzoquinoxaline-2,3-dione (NBQX) and 40 mu M -2-amino-5 phosphonopentanoic acid (AP5)]. The SGEs were composed of a negative-going change in field potential with a temporally coincident increase (0.7 +/- 0.3 mM; mean +/ SE) in extracellular K+ ([K+]o) and an alkaline transient (0.01-0.08 units) in extracellular pH (pHo) in stratum radiatum of the area CA1. Simultaneous intracellular recordings showed a triphasic hyperpolarization-depolarization-late hyperpolarization response in pyramidal cells. Application of pentobarbital sodium (PB, 100 mu M) decreased the interval between SGEs from a mean value of 35 to approximately 20 s and shortened the period of refractoriness of stimulus evoked propagating events. This was accompanied by an increase in the amplitude of the field potential response of the [K+]o and the pHo shifts and of the depolarizing phase of the pyramidal-cell response. The SGEs were completely blocked by the gamma-aminobutyric acid-A (GABAA) receptor antagonist, picrotoxin (PiTX; 100 mu M). The amplitudes of the negative-going field potential and of the depolarizing phase of the pyramidal-cell response as well as the ionic shifts associated with SGEs were strongly suppressed in the nominal absence of CO2/HCO 3. There was a five-fold increase in the interevent interval, and propagating SGEs could not be evoked by stimuli given at intervals shorter than approximately 2-3 min. Exposure to inhibitors of carbonic anhydrase, benzolamide (BA; 10 micro M) or ethoxyzolamide (EZA; 50 mu M) fully blocked the alkaline pHo transients and turned them into acid shifts. The poorly membrane-permeant BA had no discernible effect on the other components of the SGEs, but application of EZA had effects reminiscent to those of CO2/HCO-3-free medium. Addition of the GABAA receptor permeant weak-acid anion, formate (20 mM) reestablished the SGEs that were first suppressed by exposure to the CO2/HCO-3-free medium. No SGEs were seen in the presence of a similar concentration of the GABAA receptor-impermeant anion propionate. Unlike the alkaline transients associated with HCO-3-driven SGEs, those supported by formate were not blocked by BA. The present data suggest that an inward current carried by bicarbonate is necessary for the generation of SGEs and that the GABAA receptor-mediated excitatory coupling among GABAergic interneurons is essentially dependent on the availability of intracellular bicarbonate. PMID- 9356409 TI - Inwardly rectifying and Ca2+-permeable AMPA-type glutamate receptor channels in rat neocortical neurons. AB - Inwardly rectifying and Ca2+-permeable AMPA-type glutamate receptor channels in rat neocortical neurons. J. Neurophysiol. 78: 2592-2605, 1997. Current-voltage (I V) relations and Ca2+ permeability of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)type glutamate receptor channels were investigated in neurons of rat neocortex by using the whole cell patch-clamp technique in brain slices. To activate AMPA receptor channels, kainate was used as a nondesensitizing agonist. A patch pipette was filled with solution containing 100 mu M spermine to maintain the inward rectification of Ca2+-permeable AMPA receptor channels. Three types of responses to kainate were observed: type I response with outwardly rectifying I-V relation, type II response with I-V relation of marked inward rectification, and intermediate response with I-V relation of weaker inward rectification. Neurons with type I, type II and intermediate I-V relations were referred to as type I, type II, and intermediate neurons, respectively. Of a total of 223 recorded cells, 90 (40.4%) were type I, 129 (57.8%) intermediate, and 4 (1.8%) type II neurons. Properties of AMPA receptor channels were examined in the former two types of neurons. The value of PCa:PCs, the ratio of the permeability coefficients of Ca2+ and Cs+, was estimated from the reversal potentials of kainate responses in the outside-out patches bathed in Na+-free solution containing 100 mM Ca2+ according to the constant-field equation. They ranged from 0.05 to 0.10 (0.08 +/- 0. 02, mean +/- SD, n = 8) for type I neurons and from 0.14 to 1.29 (0. 60 +/- 0.37, n = 11) for the intermediate neurons. There was a close correlation between the inward rectification and the Ca2+ permeability in AMPA receptor channels in these neurons. Intermediate neurons stained with biocytin were nonpyramidal cells with ellipsoidal-shaped somata. Type I neurons had either triangular- or ellipsoidal shaped somata. Excitatory postsynaptic currents (EPSCs) recorded in both type I and intermediate neurons had 6-cyano-7-nitroquinoxaline-2,3-dione-sensitive fast and -2-amino-5-phosphonovalerate-sensitiveslow components. The I-V relation of the fast component exhibited inward rectification in the intermediate neuron, whereas that in the type I neuron showed slight outward rectification. The fast component of EPSCs in the intermediate neuron was suppressed more prominently (to 56 +/- 15% of the control, n = 12) than that in the type I neuron (to 78 +/- 6% of the control, n = 6) by bath application of 1 mM spermine. These results indicate that inwardly rectifying and Ca2+-permeable AMPA receptor channels are expressed in a population of neurons of rat neocortex and are involved in excitatory synaptic transmission. PMID- 9356410 TI - Converging inputs to the entorhinal cortex from the piriform cortex and medial septum: facilitation and current source density analysis. AB - Converging inputs to the entorhinal cortex from the piriform cortex and medial septum: facilitation and current source density analysis. J. Neurophysiol. 78: 2602-2615, 1997. The entorhinal cortex receives sensory inputs from the piriform cortex and modulatory inputs from the medial septum. To examine short-term synaptic facilitation effects in these pathways, current source density (CSD) analysis was used first to localize the entorhinal cortex membrane currents, which generate field potentials evoked by stimulation of these afferents. Field potentials were recorded at 50-micron intervals through the medial entorhinal cortex in urethan-anesthetized rats and the one-dimensional CSD was calculated. Piriform cortex stimulation evoked a surface-negative, deep-positive field potential component in the entorhinal cortex with mean onset and peak latencies of 10.4 and 18.4 ms. The component followed brief 100-Hz stimulation, consistent with a monosynaptic response. CSD analysis linked the component to a current sink, which often began in layer I before peaking in layer II. A later, surface positive field potential component peaked at latencies near 45 ms and was associated with a current source in layer II. Medial septal stimulation evoked positive and negative field potential components which peaked at latencies near 7 and 16 ms, respectively. A weaker and more prolonged surface-negative, deep positive component peaked at latencies near 25 ms. The early components were generated by currents in the hippocampal formation, and the late surface-negative component was generated by currents in layers II to IV of the entorhinal cortex. Short-term facilitation effects in conscious animals were examined using electrodes chronically implanted near layer II of the entorhinal cortex. Paired pulse stimulation of the piriform cortex at interpulse intervals of 30 and 40 ms caused the largest facilitation (248%) of responses evoked by the second pulse. Responses evoked by medial septal stimulation also were facilitated maximally (59%) by a piriform cortex conditioning pulse delivered 30-40 ms earlier. Paired pulse stimulation of the medial septum caused the largest facilitation (149%) at intervals of 70 ms, but piriform cortex evoked responses were facilitated maximally (46%) by a septal conditioning pulse 100-200 ms earlier. Frequency potentiation effects were maximal during 12- to 18-Hz stimulation of either the piriform cortex or medial septum. Occlusion tests suggested that piriform cortex and medial septal efferents activate the same neurons. The CSD analysis results show that evoked field potential methods can be used effectively in chronically prepared animals to examine synaptic responses in the converging inputs from the piriform cortex and medial septum to the entorhinal cortex. The short-term potentiation phenomena observed here suggest that low-frequency activity in these pathways during endogenous oscillatory states may enhance entorhinal cortex responsivity to olfactory inputs. PMID- 9356411 TI - Stochastic threshold characterization of the intensity of active channel dynamical action potential generation. AB - Stochastic threshold characterization of the intensity of active channel dynamical action potential generation. J. Neurophysiol. 78: 2616-2630, 1997. This paper develops a stochastic intensity description for action potential generation formulated in terms of stochastic processes, which are direct analogues of the physiological processes of the pre- and postsynaptic complex of the cochlear nerve: 1) neurotransmitter release is modeled as an inhomogeneous Poisson counting process with release intensity mu t, 2) the excitatory postsynaptic conductance (EPSC) process is modeled as a marked, linearly filtered Poisson process resulting from the linear superposition of standard shaped postsynaptic conductances of size G, and 3) action potential generation is modeled as resulting from the EPSC exceeding a random threshold determined by active channel dynamics of the Hodgkin-Huxley type. The random threshold is defined to be the least upper bound in the size of a standard-shaped neurotransmitter release injected at time t given the previous action potential time and the number of releases occurring in a short preconditioning time increment. The action potential process is modeled as a self-exciting point process with stochastic intensity resulting from the probability that the random threshold process crosses the threshold in some small time increment that is a function of time since previous action potential, release intensity, and the probability that a single synaptic event exceeds the stochastic threshold. The stochastic intensity model is consistent with a direct simulation of the nonlinear Hodgkin-Huxley differential equations over a variety of parameters for the vesicle release intensity, vesicle size, vesicle duration, and temperatures. Results are presented showing that the regularity properties seen in the vestibular primary afferent in the lizard, Calotes versicolor, associated with a slow-to-activate potassium channel resulting in a long afterhyperpolarization can be accommodated directly by the stochastic intensity description. The stimulus dependence of the model is attributed to synaptic transmission and the probabilistic nature to the threshold conductance process, which is dependent upon the EPSC process. The stochastic intensity is seen to have a form consistent with the phenomenologically based Siebert-Gaumond model, a stimulus-related function of time multiplied by a refractory-related function of time since previous action potential. PMID- 9356412 TI - Properties of carbachol-induced oscillatory activity in rat hippocampus. AB - Properties of carbachol-induced oscillatory activity in rat hippocampus. J. Neurophysiol. 78: 2631-2640, 1997. The recent resurgence of interest in carbachol oscillations as an in vitro model of theta rhythm in the hippocampus prompted us to evaluate the circuit mechanisms involved. In extracellular recordings, a regularly spaced bursting pattern of field potentials was observed in both CA3 and CA1 subfields in the presence of carbachol. Removal of the CA3 region abolished oscillatory activity observed in CA1, suggesting that the oscillatory generator is located in CA3. An alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor antagonist, 6,7-dinitroquinoxaline-2,3 dione (DNQX), blocked carbachol oscillations, indicating that AMPA receptor mediated synaptic currents are necessary for the population oscillation. Moreover, the spread of oscillatory activity into CA1 required intact N-methyl- aspartate receptors. These data are more consistent with epileptiform bursting than with theta rhythm described in vivo. In the presence of carbachol, individual CA3 pyramidal cells exhibited a slow, rhythmic intrinsic oscillation that was not blocked by DNQX and that was enhanced by membrane hyperpolarization. We hypothesize that this slower oscillation is the fundamental oscillator that participates in triggering the population oscillation by exciting multiple synaptically connected CA3 neurons. gamma-aminobutyric acid-A (GABAA) receptors are not necessary for carbachol to elicit synchronous CA3 field events but are essential to the bursting pattern observed. Neither GABAB nor metabotropic glutamate receptors appear to be necessary for carbachol oscillations. However, both nicotinic and M1 and M3 muscarinic cholinergic receptors contribute to the generation of this activity. These results establish the local circuit elements and neurotransmitter receptors that contribute to carbachol-induced oscillations and indicate that carbachol-induced oscillations are fundamentally distinct from theta rhythm in vivo. PMID- 9356413 TI - Innervation territories of mechanically activated C nociceptor units in human skin. AB - Innervation territories of mechanically activated C nociceptor units in human skin. J. Neurophysiol. 78: 2641-2648, 1997. Innervation territories of single mechanically activated C nociceptors in the skin of the leg and foot were explored in normal human subjects. Microneurographic recordings were obtained in the peroneal nerve from 70 mechano-heat responsive (CMH) and 7 mechano-(but not heat) responsive (CM) units. Units were identified by their constant long-latency response to intracutaneous electrical stimulation of their terminals. Responsiveness to mechanical, heat, or transcutaneous electrical stimuli was verified by transient slowing of conduction velocity after activation by such stimuli. We determined their thresholds to mechanical stimuli (mean 33.7 mN, median 30 mN, range 3-750 mN) and heat (mean 42.5 degrees C, median 42.5 degrees C, range 37-49 degrees C). Most mechano-receptive fields (mRFs) were found on the foot dorsum (60 units) and some on the lower leg (14 units) and toes (3 units). Most units had one continuous mRF, but 10 units had more complex fields. Areas of mRFs mapped with a von Frey filament (750 mN) ranged from 10 to 363 mm2 (mean, 106 mm2). The mRFs were oval or irregularly shaped with greatest diameters ranging from 3 to 45 mm. Mean areas of mRFs were largest on the lower leg (198 mm2), smaller on the foot dorsum (88 mm2), and smallest on the toes (35 mm2). Forty-nine of the 77 units had identical mRFs and electro-receptive fields (eRFs). Twenty-six units had larger eRFs than mRFs, whereas the opposite was found for two units only. Areas of eRFs ranged from 16 to 511 mm2 (mean 121 mm2). An estimate of the innervation density based on the present data and the presumed number of C fibers in cutaneous fascicles of the peroneal nerve suggests a considerable overlap of nociceptive endings in the skin. Such overlapping nociceptor innervation in the skin allows for substantial spatial summation in response to punctate noxious stimuli, which may be a prerequisite for high accuracy in localizing painful events from a C-fiber input. The reduction in size of innervation territories distally allows for finer discrimination of spatial dimensions of noxious stimuli distally as compared with proximal regions of the extremities. Mean maximal diameters of the mechano-receptive fields of CMH and CM units on the lower leg (22.3 mm) and foot (15.3 mm) are of similar size as the radius of axon reflex flares evoked by noxious mechanical stimuli in these regions. PMID- 9356414 TI - Effects of Pb2+ on delayed-rectifier potassium channels in acutely isolated hippocampal neurons. AB - Effects of Pb2+ on delayed-rectifier potassium channels in acutely isolated hippocampal neurons. J. Neurophysiol. 78: 2649-2654, 1997. The effects of Pb2+ on delayed-rectifier potassium currents were studied in acutely isolated hippocampal neurons (CA1 neurons, CA3 neurons, granule cells) from the guinea pig using the patch-clamp technique in the whole cell configuration. Pb2+ in micromolar concentrations decreased the potassium currents in a voltage-dependent manner, which appeared as a shift of the current-voltage relation to positive potentials. The effect was reversible after washing. The concentration-responsiveness measured in CA1 neurons revealed an IC50 value of 30 mu mol/l at a potential of 30 mV. The half-maximal shift of the current-voltage relation was reached at 33 mu mol/l and the maximal obtainable shift was 13.4 mV. For the different types of hippocampal neurons, the shift of the current-voltage relation was distinct and was 7.9 mV in CA1 neurons, 13.7 mV in CA3 neurons, and 14.2 mV in granule cells with 50 micro mol/l Pb2+. The effects described here of Pb2+ on the potassium currents in hippocampal neurons and the differences between the types of hippocampal neurons correspond with the known properties and distributions of cloned potassium channels found in the hippocampus. As a whole, our results demonstrate that Pb2+ in micromolar concentration is a voltage-dependent, reversible blocker of delayed-rectifier potassium currents of hippocampal neurons. This effect has to be taken into consideration as a possible contributing mechanism for the neurological symptoms of enhanced brain activity seen during Pb2+ intoxication. PMID- 9356415 TI - Independent coding of wind direction in cockroach giant interneurons. AB - Independent coding of wind direction in cockroach giant interneurons. J. Neurophysiol. 78: 2655-2661, 1997. In this study we examined the possible role of cell-to-cell interactions in the localization processing of a wind stimulus by the cockroach cercal system. Such sensory processing is performed primarily by pairs of giant interneurons (GIs), a group of highly directional cells. We have studied possible interactions among these GIs by comparing the wind sensitivity of a given GI before and after removing another GI with the use of photoablation. Testing various combinations of GI pairs did not reveal any suprathreshold interactions. This was true for all unilateral GI pairs on the left or right side as well as all the bilateral GI pairs (left and right homologues). Those experiments in which we were able to measure synaptic activity did not reveal subthreshold interactions between the GIs either. We conclude that the GIs code independently for a given wind direction without local GI-GI interactions. We discuss the possible implications of the absence of local interactions on information transfer in the first station of the escape circuit. PMID- 9356416 TI - Response sensitivity and voltage gain of the rod- and cone-bipolar cell synapses in dark-adapted tiger salamander retina. AB - Response sensitivity and voltage gain of the rod- and cone-bipolar cell synapses in dark-adapted tiger salamander retina. J. Neurophysiol. 78: 2662-2673, 1997. Rods, cones, and bipolar cells were recorded in superfused, flat-mounted isolated retinas of the larval tiger salamander, Ambystoma tigrinum, under dark-adapted conditions. Voltage responses of 24 rods, 15 cones, and 41 bipolar cells in dark adapted retinas to 500 nm light steps of various intensities were listed and fitted with hyperbolic functions, and their step sensitivities and relative sensitivities (log sigma) were estimated. In the linear response-intensity ranges, the step sensitivity of rods, SS(rod), is -1.0 mV photon-1 micron2 s or 0.034 mV Rh*-1 s rod and that of the cones, SS(cone), is approximately 0. 00146 mV photon-1 micron2 s or 0.000048 mV Rh*-1 s rod. The rod and cone responses were relatively homogenous with little variations in response amplitude and sensitivity. In contrast, bipolar cell responses were heterogenous with large variations in response amplitude and sensitivity. The maximum response amplitude of bipolar cells varied from 5 to 25 mV, and the relative response sensitivity (log sigma) varied >6 log units (-8.11 to -2.32). The step sensitivity of bipolar cells in the linear response-intensity range varied from 0.0000438 to 51.82 mV photon-1 micron2 s. Bipolar cells in dark-adapted tiger salamander retinas fell into two groups according to their relative sensitivities with very few cells falling in the intermediate light intensity region. The mixed bipolar cells (DBCM and HBCM) exhibited relative response sensitivity ranged from -8.11 to -5.54, and step sensitivity ranged from 1.22 to 51.82 mV photon-1 micron2 s. The cone-driven bipolar cells (DBCC and HBCC) exhibited relative response sensitivity ranged from -3.45 to -2.32, and step sensitivity ranged from 0.0000438 to 0. 00201 mV photon 1 micron2 sec. The chord voltage gain of the rod-DBCM or rod-HBCM synapses near the rod dark membrane potential ranged from 1.14 to 48.43 and that of the cone DBCC or cone-HBCC synaptic gain near the cone dark membrane potential ranged from 0.03 to 1.38. The highest voltage gains were found near the rod or cone dark membrane potentials. By the use of linear subtraction method, we studied the synaptic inputs from cones to five mixed bipolar cells, and the voltage gains of the cone synapses in each of the bipolar cells were very close to the voltage gain of the rod synapses. This result suggests that although the responses of mixed bipolar cells are mediated mainly by rods when lights of short and medium wavelengths are used, their responses to long wavelength lights (>650 nm) are mediated by both rods and cones with comparable synaptic gains. Functional roles of the mixed and cone-driven bipolar cells in information processing in dark adapted retinas are discussed. PMID- 9356418 TI - Mechanisms of potentiation by calcium-calmodulin kinase II of postsynaptic sensitivity in rat hippocampal CA1 neurons. AB - Mechanisms of potentiation by calcium-calmodulin kinase II of postsynaptic sensitivity in rat hippocampal CA1 neurons. J. Neurophysiol. 78: 2682-2692, 1997. Preactivated recombinant alpha-calcium-calmodulin dependent multifunctional protein kinase II (CaMKII*) was perfused internally into CA1 hippocampal slice neurons to test the effect on synaptic transmission and responses to exogenous application of glutamate analogues. After measurement of baseline transmission, internal perfusion of CaMKII* increased synaptic strength in rat hippocampal neurons and diminished the fraction of synaptic failures. After measurement of baseline responses to applied transmitter, CaMKII* perfusion potentiated responses to kainate but not responses to N-methyl--aspartate. Internal perfusion of CaMKII*potentiated the maximal effect of kainate. Potentiation by CaMKII* did not change the time course of responses to kainate, whereas increasing response size by pharmacologically manipulating desensitization or deactivation rate constants significantly altered the time course of responses. Nonstationary fluctuation analysis of responses to kainate showed a decrease in the coefficient of variation after potentiation by CaMKII*. These data support the hypothesis that CaMKII increases postsynaptic responsiveness by increasing the available number of active alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate channels and suggests that a similar process may occur during the expression of long-term potentiation. PMID- 9356417 TI - Ca2+-activated K+ currents in rat locus coeruleus neurons induced by experimental ischemia, anoxia, and hypoglycemia. AB - Ca2+-activated K+ currents in rat locus coeruleus neurons induced by experimental ischemia, anoxia, and hypoglycemia. J. Neurophysiol. 78: 2674-2681, 1997. The effects of metabolic inhibition on membrane currents and N-methyl--aspartic acid (NMDA)-induced currents were investigated in dissociated rat locus coeruleus (LC) neurons by using the nystatin perforated patch recording mode under voltage-clamp conditions. Changes in the intracellular Ca2+ concentration ([Ca2+]i) during the metabolic inhibition were also investigated by using the microfluometry with a fluorescent probe, Indo-1. Removal of both the oxygen and glucose (experimental ischemia), deprivation of glucose (hypoglycemia), and a blockade of electron transport by sodium cyanide (NaCN) or a reduction of the mitochondrial membrane potential with carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone(FCCP) as experimental anoxia all induced a slowly developing outward current (IOUT) at a holding potential of -40 mV. The application of 10(-4) M NMDA induced a rapid transient peak and a successive steady state inward current and a transient outward current immediately after washout. All treatments related to metabolic inhibition increased the NMDA-induced outward current(INMDA-OUT) and prolonged the one-half recovery time of INMDA-OUT. The reversal potentials of both IOUT and INMDA-OUT were close to the K+ equilibrium potential (EK) of -82 mV. Either charybdotoxin or tolbutamide inhibited the IOUT and INMDA-OUT, suggesting the contribution of Ca2+-activated and ATP-sensitive K+ channels, even though the inhibitory effect of tolbutamide gradually diminished with time. Under the metabolic inhibition, the basal level of [Ca2+]i was increased and the one-half recovery time of the NMDA-induced increase in [Ca2+]i was prolonged. The IOUT induced by NaCN was inhibited by a continuous treatment of thapsigargin but not by ryanodine, indicating the involvement of inositol 1,4, 5-trisphosphate (IP3) induced Ca2+ release (IICR) store. These findings suggest that energy deficiency causes Ca2+ release from the IICR store and activates continuous Ca2+-activated K+ channels and transient ATP-sensitive K+ channels in acutely dissociated rat LC neurons. PMID- 9356419 TI - A fast synaptic potential mediated by NMDA and non-NMDA receptors. AB - A fast synaptic potential mediated by NMDA and non-NMDA receptors. J. Neurophysiol. 78: 2693-2706, 1997. Excitatory synaptic transmission in the CNS often is mediated by two kinetically distinct glutamate receptor subtypes that frequently are colocalized, the N-methyl--aspartate (NMDA) and non-NMDA receptors. Their synaptic currents are typically very slow and very fast, respectively. We examined the pharmacological and physiological properties of chemical excitatory transmission at the mixed electrical and chemical synapses between auditory afferents and the goldfish Mauthner cell, in vivo. Previous physiological data have suggested the involvement of glutamate receptors in this fast excitatory postsynaptic potential (EPSP), the chemical component of which decays with a time constant of <2 ms. We demonstrate here that the pharmacological and voltage-dependent characteristics of the synaptic currents are consistent with glutamatergic transmission and that both NMDA and non-NMDA receptors are involved. The two components surprisingly exhibit quite similar kinetics even at resting potential, with the NMDA response being only slightly slower. Due to its fast kinetics and characteristic voltage dependence, NMDA receptor-mediated transmission at these first-order synapses contributes significantly to paired pulse and frequency-dependent facilitation of successive fast EPSPs during high-frequency repetitive firing, a presynaptic impulse pattern that induces activity-dependent homosynaptic changes in both electrical and chemical transmission. Thus NMDA receptor kinetics in this intact preparation are suited to its functional requirements, namely speed of information transmission and the ability to trigger changes in synaptic efficacy. PMID- 9356420 TI - Attenuation of paired-pulse facilitation associated with synaptic potentiation mediated by postsynaptic mechanisms. AB - Attenuation of paired-pulse facilitation associated with synaptic potentiation mediated by postsynaptic mechanisms. J. Neurophysiol. 78: 2707-2716, 1997. The relationship between paired-pulse facilitation (PPF) and synaptic potentiation induced by various protocols and their cellular and molecular mechanisms were examined by extracellular field potential and current- or voltage-clamp recordings at CA1 synapses in rat hippocampal slices. Microelectrodes were used for both intracellular recordings and injections of modulators of calcium (Ca2+) and Ca2+/calmodulin (CaM) signaling pathways into postsynaptic neurons. Basal synaptic transmission was not accompanied by changes in PPF. Tetanic stimulation induced long-term potentiation (LTP) of synaptic transmission and attenuated PPF. Experiments stimulating two independent Schaffer collateral/commisural(S/C) pathways showed that PPF attenuation and tetanus-LTP were pathway specific. Postsynaptic injections of pseudosubstrate inhibitors of CaM-dependent protein kinase II and protein kinase C (CaM-KII/PKC), [Ala286]CaMKII286-302 plus PKC19 31, almost completely attenuated tetanus-LTP and reversed PPF attenuation but did not affect synaptic transmission and PPF under basal conditions. Postsynaptic injections of heparin and dantrolene (inhibitors of IP3 and ryanodine receptors at intracellular Ca2+ stores) prevented tetanus-LTP induction and PPF attenuation. Postsynaptic injections of calcineurin (CaN) inhibitors, CaN autoinhibitory peptide (CaN-AIP) or FK-506, enhanced synaptic transmission and decreased PPF. CaN-inhibited synaptic potentiation and PPF attenuation were unaffected by (-)-a-Amino-5-phosphonopentanoic, but blocked by coinjecting 1, 2 bis(2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, heparin plus dantrolene, calmodulin-binding peptide, or [Ala286]CaMKII281-302 plus PKC19-31. PPF attenuation associated with tetanus-LTP or CaN-inhibited synaptic potentiation resulted from smaller increases in the potentiation of the second synaptic responses (R2) compared with the potentiation of the first responses (R1). Our results indicate that PPF attenuation is associated with synaptic potentiation mediated by postsynaptic mechanisms, and postsynaptic Ca2+/CaM signaling pathways play a dual role in synaptic plasticity. CaN activity limits synaptic transmission under basal conditions, whereas the activation of Ca2+-dependent protein kinases enhances synaptic transmission and attenuates PPF at central synapses. PMID- 9356421 TI - Coding for auditory space in the nucleus of the brachium of the inferior colliculus in the ferret. AB - Coding for auditory space in the nucleus of the brachium of the inferior colliculus in the ferret. J. Neurophysiol. 78: 2717-2731, 1997. The nucleus of the brachium of the inferior colliculus (BIN) projects topographically to the deeper layers of the superior colliculus (SC), which contain a two-dimensional map of auditory space. In this study, we have used broadband stimuli presented in the free field to investigate how auditory space is represented in the BIN of the ferret. Response latencies and temporal firing patterns were comparable with those in the SC, and both properties showed some variation with stimulus location. We obtained spatial response profiles at two sound levels (5-15 and 25 35 dB above unit threshold). A large proportion of azimuth profiles (41% in the suprathreshold condition, 80% in the near-threshold condition) presented a single peak, indicating that they were tuned to single regions in space. For some of these units, the preferred speaker position varied considerably with sound level. The remaining units showed predominantly either broad "hemifield" or spatially ambiguous "bilobed" response profiles. At suprathreshold sound levels, the preferred azimuths of the tuned cells were ordered topographically along the rostrocaudal axis of the BIN, although this representation is considerably more scattered than that in the SC. In contrast to the SC, we observed no systematic variation in the distribution of near-threshold best azimuths, which were instead concentrated around the interaural axis in the contralateral hemifield. The azimuth tuning of individual units in the BIN was generally broader at both sound levels than that in the SC. Many units also were tuned for the elevation of the sound source (48% for supra-, 77% for near-threshold stimulation), but there was no evidence for topographic order in the distribution of preferred elevations within the BIN. These results suggest that the BIN sends inputs to the SC that are already selective for sound azimuth and elevation and that show some degree of topographic order for sound azimuth. These inputs then presumably are sharpened and their topography refined by a mechanism that is likely to involve convergence of other inputs and activity-dependent fine tuning of terminal connections, to result in a precise two-dimensional map of auditory space in the SC. PMID- 9356422 TI - Visual response properties and visuotopic representation in the newborn monkey superior colliculus. AB - Visual response properties and visuotopic representation in the newborn monkey superior colliculus. J. Neurophysiol. 78: 2732-2741, 1997. Visually responsive neurons were recorded in the superior colliculus (SC) of the newborn rhesus monkey. The receptive fields of these neurons were larger than those in the adult, but already were organized into a well-ordered map of visual space that was very much like that seen in mature animals. This included a marked expansion of the representation of the central 10 degrees of the visual field and a systematic foveal to peripheral increase in receptive field size. Although newborn SC neurons had longer response latencies than did their adult counterparts, they responded vigorously to visual stimuli and exhibited many visual response properties that are characteristic of the adult. These included surround inhibition, within-field spatial summation, within-field spatial inhibition, binocularity, and an adult-like ocular dominance distribution. As in the adult, SC neurons in the newborn preferred a moving visual stimulus and had adult-like selectivities for stimulus speed. The developmentally advanced state of the functional circuitry of the newborn monkey SC contrasts with the comparative immaturity of neurons in its visual cortex. It also contrasts with observations on the state of maturation of the newborn SC in other developmental models (e.g., cat). The observation that extensive visual experience is not necessary for the development of many adult-like SC response properties in the monkey SC may help explain the substantial visual capabilities shown by primates soon after birth. PMID- 9356423 TI - Absence of a prevalent laminar distribution of IPSPs in association cortical neurons of cat. AB - Absence of a prevalent laminar distribution of IPSPs in association cortical neurons of cat. J. Neurophysiol. 78: 2742-2753, 1997. The depth distribution of inhibitory postsynaptic potentials (IPSPs) was studied in cat suprasylvian (association) cortex in vivo. Single and dual simultaneous intracellular recordings from cortical neurons were performed in the anterior part of suprasylvian gyrus (area 5). Synaptic responses were obtained by stimulating the suprasylvian cortex, 2-3 mm anterior to the recording site, as well as the thalamic lateral posterior (LP) nucleus. Neurons were recorded from layers 2 to 6 and were classified as regular spiking (RS, n = 132), intrinsically bursting (IB, n = 24), and fast spiking (FS, n = 4). Most IB cells were located in deep layers (below 0.7 mm, n = 19), but we also found some IB cells more superficially (between 0.2 and 0.5 mm, n = 5). Deeply lying corticothalamic neurons were identified by their antidromic invasion on thalamic stimulation. Neurons responded with a combination of excitatory postsynaptic potentials (EPSPs) and IPSPs to both cortical and thalamic stimulation. No consistent relation was found between cell type or cell depth and the amplitude or duration of the IPSPs. In response to thalamic stimulation, RS cells had IPSPs of 7.9 +/- 0.9 (SE) mV amplitude and 88.9 +/- 6.4 ms duration. In IB cells, IPSPs elicited by thalamic stimulation had 7.4 +/- 1.3 mV amplitude and 84.7 +/- 14.3 ms duration. The differences between the two (RS and IB) groups were not statistically significant. Compared with thalamically elicited inhibitory responses, cortical stimulation evoked IPSPs with higher amplitude (12.3 +/- 1.7 mV) and longer duration (117 +/- 17.3 ms) at all depths. Both cortically and thalamically evoked IPSPs were predominantly monophasic. Injections of Cl- fully reversed thalamically as well as cortically evoked IPSPs and revealed additional late synaptic components in response to cortical stimulation. These data show that the amount of feed forward and feedback inhibition to cat's cortical association cells is not orderly distributed to distinct layers. Thus local cortical microcircuitry goes beyond the simplified structure determined by cortical layers. PMID- 9356424 TI - Monaural spectral contrast mechanism for neural sensitivity to sound direction in the medial geniculate body of the cat. AB - Monaural spectral contrast mechanism for neural sensitivity to sound direction in the medial geniculate body of the cat. J. Neurophysiol. 78: 2754-2771, 1997. Central auditory neurons vary in sound direction sensitivity. Insensitive cells discharge well to all sound source directions, whereas sensitive cells discharge well to certain directions and poorly to others. High-frequency neurons in the latter group are differentially sensitive to binaural and monaural directional cues present in broadband noise (BBN). Binaural directional (BD) cells require binaural stimulation for directional sensitivity; monaural directional (MD) cells are sensitive to the direction of monaural stimuli. A model of MD sensitivity was tested using single-unit responses. The model assumes that MD cells derive directional sensitivity from pinna-derived spectral cues (head related transfer function, HRTF). This assumption was supported by the similarity of effects that pinna orientation produces on locations of HRTF patterns and on locations of MD cell azimuth function peaks and nulls. According to the model, MD neurons derive directional sensitivity by use of excitatory/inhibitory antagonism to compare sound pressure in excitatory and inhibitory frequency domains, and a variety of observations are consistent with this idea. 1) Frequency response areas of MD cells consist of excitatory and inhibitory domains. MD cells exhibited a higher proportion of multiple excitatory domains and narrower excitatory frequency domains than BD cells, features that may reflect specialization for spectral dependent directional sensitivity. 2) MD sensitivity requires sound pressure in excitatory and inhibitory frequency domains. Directional sensitivity was evaluated using stimuli with frequency components confined exclusively to excitatory domains (E-only stimuli) or distributed in both excitatory and inhibitory domains (E/I stimuli). Each of 13 MD cells that were tested exhibited higher directional sensitivity to E/I than to E-only stimuli; most MD cells exhibited relatively low directional sensitivity when frequency components were confined exclusively to excitatory domains. 3) MD sensitivity derives from excitatory/inhibitory antagonism (spectral inhibition). Comparison of responses to best frequency and E/I stimuli provided strong support for spectral inhibition. Although spectral facilitation conceivably could contribute to directional sensitivity with direction-dependent increases in response, the results did not show this to be a significant factor. 4) Direction-dependent decreases in responsiveness to BBN reflect increased sound pressure in inhibitory relative to excitatory frequency domains. This idea was tested using the strength of two-tone inhibition, which is a function of stimulus levels in inhibitory relative to excitatory frequency domains. The finding that two-tone inhibition was stronger at directions where BBN responses were minimal than at directions where they were maximal supports the model. PMID- 9356425 TI - Direction selectivity of synaptic potentials in simple cells of the cat visual cortex. AB - Direction selectivity of synaptic potentials in simple cells of the cat visual cortex. J. Neurophysiol. 78: 2772-2789, 1997. The direction selectivity of simple cells in the visual cortex is generated at least in part by nonlinear mechanisms. If a neuron were spatially linear, its responses to moving stimuli could be predicted accurately from linear combinations of its responses to stationary stimuli presented at different positions within the receptive field. In extracellular recordings, this has not been found to be the case. Although the extracellular experiments demonstrate the presence of a nonlinearity, the cellular process underlying the nonlinearity, whether an early synaptic mechanism such as a shunting inhibition or simply the spike threshold at the output, is not known. To differentiate between these possibilities, we have recorded intracellularly from simple cells of the intact cat with the whole cell patch technique. A linear model of direction selectivity was used to analyze the synaptic potentials evoked by stationary sine-wave gratings. The model predicted the responses of cells to moving gratings with considerable accuracy. The degree of direction selectivity and the time course of the responses to moving gratings were both well matched by the model. The direction selectivity of the synaptic potentials was considerably smaller than that of the intracellularly recorded action potential, indicating that a nonlinear mechanism such as threshold enhances the direction selectivity of the cell's output over that of its synaptic inputs. At the input stage, however, the cells apparently sum their synaptic inputs in a highly linear fashion. A more constrained test of linearity of synaptic summation based on principal component analysis was applied to the responses of direction-selective cells to stationary gratings. The analysis confirms that the summation in these cells is highly linear. The principal component analysis is consistent with a model in which direction selectivity in cortical simple cells is generated by only two subunits, each with a different receptive-field position and response time course. The response time course for each of the two subunits is derived for four analyzed cells. Each derived subunit is linear in spatial summation, suggesting that the neurons that comprise each subunit are either geniculate X-cells or receive their primary synaptic input from X-cells. The amplitude of the response of each subunit is linearly related to the contrast of the stimulus. The subunits are nonlinear in the time domain, however: the response to a stationary stimulus whose contrast is modulated sinusoidally in time is nonsinusoidal. The principal component analysis does not exclude models of direction selectivity based on more than two subunits, but such higher-order models would have to include the constraint that the extra subunits form a smooth continuum of interpolation between the properties derived from the two subunit solution. PMID- 9356426 TI - Axotomy increases the excitability of dorsal root ganglion cells with unmyelinated axons. AB - Axotomy increases the excitability of dorsal root ganglion cells with unmyelinated axons. J. Neurophysiol. 78: 2790-2794, 1997. To better understand the neuronal mechanism of neuropathic pain, the effect of axotomy on the excitability of dorsal root ganglion (DRG) cells with unmyelinated axons (C cells) was investigated. Whole cell patch-clamp recordings were performed on intact DRG cells with intact axons or with axons transected 7-12 days earlier. C cells were identified by 1) soma size, 2) action potential morphology, 3) conduction velocity, and 4) in some cases, injection of Fast Blue into the injured nerve fibers. Axotomy reduced (more negative) action potential threshold but did not significantly change resting membrane potential, action potential duration, or maximal depolarization rate. Axotomy significantly increased the peak sodium current measured under voltage-clamp conditions. In Fast Blue-labeled (injured) cells, the tetrodotoxin (TTX)-sensitive current was enhanced while the TTX-resistant current was reduced. These results suggest that axotomy increased the excitability of C cells, possibly because of a preferential increase in expression of TTX-sensitive sodium currents. PMID- 9356427 TI - Increase of extracellular dopamine in primate prefrontal cortex during a working memory task. AB - Increase of extracellular dopamine in primate prefrontal cortex during a working memory task. J. Neurophysiol. 78: 2795-2798, 1997. The dopamine innervation of the prefrontal cortex is involved importantly in cognitive processes, such as tested in working memory tasks. However, there have been no studies directly investigating prefrontal dopamine levels in relation to cognitive processes. We measured frontal extracellular dopamine concentration using in vivo microdialysis in monkeys performing in a delayed alternation task as a typical working memory paradigm and in a sensory-guided control task. We observed a significant increase in dopamine level in the delayed alternation task as compared both with the sensory-guided control task and the basal resting level. The increase was seen in the dorsolateral prefrontal but not in the arcuate or orbitofrontal areas. The increase appeared to reflect the working memory component of the task and was observed mainly in the lip areas of principal sulcus. Although there was no significant difference in dopamine level between delayed alternation and sensory guided control tasks in the premotor area, significant increases in dopamine concentration were observed during both tasks as compared with the basal resting level, indicating the importance of premotor dopamine for the motor response itself. PMID- 9356429 TI - Epileptogenesis following neocortical trauma from two sources of disinhibition. AB - Epileptogenesis following neocortical trauma from two sources of disinhibition. J. Neurophysiol. 78: 2804-2810, 1997. Intracellular and field potential recordings were obtained from superficial and deep neurons from both intact coronal rat somatosensory slices, and from slices which had been acutely divided into a superficial strip of cortex ( approximately 450 micron from the pia) and a deep segment. Membrane properties for cells in the traumatized slices were similar to those of their counterparts in intact slices. However, synaptic hyperexcitability developed in the deep segments in which a majority of cells likely underwent dendrotomy. This hyperexcitability was manifested by epileptiform activity in 54% of traumatized slices. Measurements of fastGABAergic inhibitory strength showed these slices were disinhibited. Superficial delivery of tetrodotoxin to the upper 450 micron of intact slices led to disinhibition of fast GABAergic transmission as well as an attendant increase in excitatory postsynaptic potential strength but not epileptogenesis. Pharmacological maneuvers aimed at preventing glutamate-triggered increases in intracellular calcium [glutamate ionotropic antagonists, dantrolene, and bis-(o-aminophenoxy) N,N,N', N'-tetraacetic acid (BAPTA)-AM] showed that a 1 h treatment in these agents conferred protection against epileptogenesis. These results demonstrate that the seizure-like activity developing in deep dendrotomized cortical segments resulted from two sources of GABAergic disinhibition: the physical removal of important superficial inhibitory circuits and glutamate-triggered increases in intracellular calcium. PMID- 9356428 TI - NMDA receptor involvement in neuroplastic changes induced by neonatal capsaicin treatment in trigeminal nociceptive neurons. AB - NMDA receptor involvement in neuroplastic changes induced by neonatal capsaicin treatment in trigeminal nociceptive neurons. J. Neurophysiol. 78: 2799-2803, 1997. This study examines whether 1) the neonatal loss of C-fiber afferents results in neuroplastic changes in the mechanoreceptive field (RF) properties and spontaneous activity of nociceptive neurons in trigeminal subnucleus caudalis (medullary dorsal horn) of adult rats, and that 2) N-methyl--aspartic acid (NMDA) receptor mechanisms are involved in these neuroplastic changes. Compared with vehicle-treated (i.e., control, CON) rats, capsaicin-treated (CAP) rats showed a marked increase in neuronal spontaneous activity and RF size per se, but these neuroplastic changes could be significantly reduced by MK-801 (1 mg/kg, iv), a noncompetitive NMDA receptor antagonist; RF size and spontaneous activity remained unchanged in CON rats after MK-801 administration and in CAP rats after vehicle (saline, iv). Administration of 7-chlorokynurenic acid intrathecally (5 microgram/10 microliter), an antagonist of strychnine-insensitive glycine bindin sites on the NMDA receptor, also significantly reduced neuronal RF size and spontaneous activity in CAP rats, but not in CON rats. These data provide evidence that C-fiber afferents play a role in shaping the properties of nociceptive neurons and that the neuroplastic changes involve NMDA receptor mechanisms. PMID- 9356430 TI - Primate head-free saccade generator implements a desired (post-VOR) eye position command by anticipating intended head motion. AB - Primate head-free saccade generator implements a desired (post-VOR) eye position command by anticipating intended head motion. J. Neurophysiol. 78: 2811-2816, 1997. When we glance between objects, the brain ultimately controls gaze direction in space. However, it is currently unclear how this is allocated into separate commands for eye and head movement. To determine the role of desired final eye position commands, and their coordination with intended head movement, we trained three monkeys to make large gaze shifts while wearing opaque goggles with a monocular 8 degrees aperture. Animals eventually developed a new set of context-dependent eye-head coordination strategies, in particular expanding the head range and compressing the eye-in-head range toward the aperture (while wearing the goggles). However, when we shifted the location of the aperture to a different subsection of the normal head-free oculomotor range (by covering the original aperture and creating a new one), eye-head saccades failed to acquire visual targets, because they continued to drive the eye ultimately toward the now occluded original aperture. Even when a head-stationary saccade acquired the new aperture, subsequent head-free saccades drove the eye eccentrically toward a point that anticipated the intended head movement, such that the subsequent vestibuloocular reflex slow phase brought the eye onto the location of the original aperture. Animals could only acquire the new aperture consistently after several days of retraining. These results suggest that 1) eye-head coordination is achieved by a plastic, context-dependent neural operator that uses information about initial eye/head position and intended movement to compute desired combinations of final eye/head position and 2) acquisition of these positions involves sophisticated anticipatory compensations for subsequent movement components, akin to those observed previously in complex oral and manual behaviors. PMID- 9356431 TI - Gain adaptation of eye and head movement components of simian gaze shifts. AB - Gain adaptation of eye and head movement components of simian gaze shifts. J. Neurophysiol. 78: 2817-2821, 1997. To investigate the site of gaze adaptation in primates, we reduced the gain of large head-restrained gaze shifts made to 50 degrees target steps by jumping the target 40% backwards during a targeting saccade and then tested gain transfer to the eye- and head-movement components of head-unrestrained gaze shifts. After several hundred backstep trials, saccadic gain decreased by at least 10% in 8 of 13 experiments, which were then selected for further study. The minimum saccadic gain decrease in these eight experiments was 12.8% (mean = 18.4%). Head-unrestrained gaze shifts to ordinary 50 degrees target steps experienced a gain reduction of at least 9.3% (mean = 14.9%), a mean gain transfer of 81%. Both the eye and head components of the gaze shift also decreased. However, average head movement gain decreased much more (22.1%) than eye movement gain (9.2%). Also, peak head velocity generally decreased significantly (20%), but peak eye velocity either increased or remained constant (average increase of 5.6%). However, the adapted peak eye and head velocities were appropriate for the adapted, smaller gaze amplitudes. Similar dissociations in eye and head metrics occurred when head-unrestrained gaze shifts were adapted directly (n = 2). These results indicated that head-restrained saccadic gain adaptation did not produce adaptation of eye movement alone. Nor did it produce a proportional gain change in both eye and head movement. Rather, normal eye and head amplitude and velocity relations for a given gaze amplitude were preserved. Such a result could be explained most easily if head-restrained adaptation were realized before the eye and head commands had been individualized. Therefore, gaze adaptation is most likely to occur upstream of the creation of separate eye and head movement commands. PMID- 9356432 TI - A total system approach to sustainable pest management. AB - A fundamental shift to a total system approach for crop protection is urgently needed to resolve escalating economic and environmental consequences of combating agricultural pests. Pest management strategies have long been dominated by quests for "silver bullet" products to control pest outbreaks. However, managing undesired variables in ecosystems is similar to that for other systems, including the human body and social orders. Experience in these fields substantiates the fact that therapeutic interventions into any system are effective only for short term relief because these externalities are soon "neutralized" by countermoves within the system. Long term resolutions can be achieved only by restructuring and managing these systems in ways that maximize the array of "built-in" preventive strengths, with therapeutic tactics serving strictly as backups to these natural regulators. To date, we have failed to incorporate this basic principle into the mainstream of pest management science and continue to regress into a foot race with nature. In this report, we establish why a total system approach is essential as the guiding premise of pest management and provide arguments as to how earlier attempts for change and current mainstream initiatives generally fail to follow this principle. We then draw on emerging knowledge about multitrophic level interactions and other specific findings about management of ecosystems to propose a pivotal redirection of pest management strategies that would honor this principle and, thus, be sustainable. Finally, we discuss the potential immense benefits of such a central shift in pest management philosophy. PMID- 9356433 TI - Gatekeepers of organ growth. PMID- 9356434 TI - Archaeal chromatin: virtual or real? PMID- 9356436 TI - Effective intercellular communication distances are determined by the relative time constants for cyto/chemokine secretion and diffusion. AB - A cell's ability to effectively communicate with a neighboring cell is essential for tissue function and ultimately for the organism to which it belongs. One important mode of intercellular communication is the release of soluble cyto- and chemokines. Once secreted, these signaling molecules diffuse through the surrounding medium and eventually bind to neighboring cell's receptors whereby the signal is received. This mode of communication is governed both by physicochemical transport processes and cellular secretion rates, which in turn are determined by genetic and biochemical processes. The characteristics of transport processes have been known for some time, and information on the genetic and biochemical determinants of cellular function is rapidly growing. Simultaneous quantitative analysis of the two is required to systematically evaluate the nature and limitations of intercellular signaling. The present study uses a solitary cell model to estimate effective communication distances over which a single cell can meaningfully propagate a soluble signal. The analysis reveals that: (i) this process is governed by a single, key, dimensionless group that is a ratio of biological parameters and physicochemical determinants; (ii) this ratio has a maximal value; (iii) for realistic values of the parameters contained in this dimensionless group, it is estimated that the domain that a single cell can effectively communicate in is approximately 250 micron in size; and (iv) the communication within this domain takes place in 10-30 minutes. These results have fundamental implications for interpretation of organ physiology and for engineering tissue function ex vivo. PMID- 9356435 TI - p53-mediated protective responses to UV irradiation. PMID- 9356439 TI - A general method to design dominant negatives to B-HLHZip proteins that abolish DNA binding. AB - We describe a method to design dominant-negative proteins (D-N) to the basic helix-loop-helix-leucine zipper (B-HLHZip) family of sequence-specific DNA binding transcription factors. The D-Ns specifically heterodimerize with the B HLHZip dimerization domain of the transcription factors and abolish DNA binding in an equimolar competition. Thermal denaturation studies indicate that a heterodimer between a Myc B-HLHZip domain and a D-N consisting of a 12-amino acid sequence appended onto the Max dimerization domain (A-Max) is -6.3 kcal.mol-1 more stable than the Myc:Max heterodimer. One molar equivalent of A-Max can totally abolish the DNA binding activity of a Myc:Max heterodimer. This acidic extension also has been appended onto the dimerization domain of the B-HLHZip protein Mitf, a member of the transcription factor enhancer binding subfamily, to produce A-Mitf. The heterodimer between A-Mitf and the B-HLHZip domain of Mitf is -3.7 kcal.mol-1 more stable than the Mitf homodimer. Cell culture studies show that A-Mitf can inhibit Mitf-dependent transactivation both in acidic extension and in a dimerization-dependent manner. A-Max can inhibit Myc-dependent foci formation twice as well as the Max dimerization domain (HLHZip). This strategy of producing D-Ns may be applicable to other B-HLHZip or B-HLH proteins because it provides a method to inhibit the DNA binding of these transcription factors in a dimerization-specific manner. PMID- 9356438 TI - Trajectory of DNA in the RNA polymerase II transcription preinitiation complex. AB - By using site-specific protein-DNA photocrosslinking, we define the positions of TATA-binding protein, transcription factor IIB, transcription factor IIF, and subunits of RNA polymerase II (RNAPII) relative to promoter DNA within the human transcription preinitiation complex. The results indicate that the interface between the largest and second-largest subunits of RNAPII forms an extended, approximately 240 A channel that interacts with promoter DNA both upstream and downstream of the transcription start. By using electron microscopy, we show that RNAPII compacts promoter DNA by the equivalent of approximately 50 bp. Together with the published structure of RNAPII, the results indicate that RNAPII wraps DNA around its surface and suggest a specific model for the trajectory of the wrapped DNA. PMID- 9356440 TI - The ribosome-in-pieces: binding of elongation factor EF-G to oligoribonucleotides that mimic the sarcin/ricin and thiostrepton domains of 23S ribosomal RNA. AB - An oligoribonucleotide (a 27-mer) that mimics the sarcin/ricin (S/R) domain of Escherichia coli 23S rRNA binds elongation factor EF-G; the Kd is 6.9 microM, whereas for binding to ribosomes it is 0.7 microM. Binding saturates when EF-G and the S/R RNA are equimolar; at saturation 70% of the input RNA is in complexes with EF-G. Binding of EF-G to S/R RNA does not require GTP but is inhibited by GDP; the inhibition by GDP is overcome by GTP. The effects of mutations of the S/R domain nucleotides G2655, A2660, and G2661 suggest that EF-G recognizes the conformation of the RNA rather than the identity of the nucleotides. EF-G also binds to an oligoribonucleotide (an 84-mer) that has the thiostrepton region of 23S rRNA; however, EF-G binds independently to S/R and thiostrepton oligoribonucleotides. PMID- 9356441 TI - A dileucine motif in the C terminus of the beta2-adrenergic receptor is involved in receptor internalization. AB - The cytoplasmic C terminus of the beta2-adrenergic receptor and many other G protein-coupled receptors contains a dileucine sequence that has been implicated in endosome/lysosome targeting of diverse proteins. In the present study, we provide evidence for an essential role of this motif in the agonist-induced internalization of the beta2-adrenergic receptor. Mutation of Leu-339 and/or Leu 340 to Ala caused little changes in surface expression, ligand binding, G protein coupling, and signaling to adenylyl cyclase, when these receptors were transiently or stably expressed in CHO or HEK-293 cells. However, agonist-induced receptor internalization was markedly impaired in the L339,340A double mutant and reduced in the two single mutants. This impairment in receptor internalization was seen by using various approaches to determine internalization: binding of hydrophobic vs. hydrophilic ligands, loss of surface beta2-adrenergic receptor immunoreactivity, and immunofluorescence microscopy. The selective effects of these mutations suggest that the C-terminal dileucine motif is involved in agonist-induced internalization of the beta2-adrenergic receptor. PMID- 9356442 TI - Crystal structure of truncated human apolipoprotein A-I suggests a lipid-bound conformation. AB - The structure of truncated human apolipoprotein A-I (apo A-I), the major protein component of high density lipoprotein, has been determined at 4-A resolution. The crystals comprise residues 44-243 (exon 4) of apo A-I, a fragment that binds to lipid similarly to intact apo A-I and that retains the lipid-bound conformation even in the absence of lipid. The molecule consists almost entirely of a pseudo continuous, amphipathic alpha-helix that is punctuated by kinks at regularly spaced proline residues; it adopts a shape similar to a horseshoe of dimensions 125 x 80 x 40 A. Four molecules in the asymmetric unit associate via their hydrophobic faces to form an antiparallel four-helix bundle with an elliptical ring shape. Based on this structure, we propose a model for the structure of apo A-I bound to high density lipoprotein. PMID- 9356443 TI - RNA-peptide fusions for the in vitro selection of peptides and proteins. AB - Covalent fusions between an mRNA and the peptide or protein that it encodes can be generated by in vitro translation of synthetic mRNAs that carry puromycin, a peptidyl acceptor antibiotic, at their 3' end. The stable linkage between the informational (nucleic acid) and functional (peptide) domains of the resulting joint molecules allows a specific mRNA to be enriched from a complex mixture of mRNAs based on the properties of its encoded peptide. Fusions between a synthetic mRNA and its encoded myc epitope peptide have been enriched from a pool of random sequence mRNA-peptide fusions by immunoprecipitation. Covalent RNA-peptide fusions should provide an additional route to the in vitro selection and directed evolution of proteins. PMID- 9356444 TI - Atomic structure of a thermostable subdomain of HIV-1 gp41. AB - Infection by HIV-1 involves the fusion of viral and cellular membranes with subsequent transfer of viral genetic material into the cell. The HIV-1 envelope glycoprotein that mediates fusion consists of the surface subunit gp120 and the transmembrane subunit gp41. gp120 directs virion attachment to the cell-surface receptors, and gp41 then promotes viral-cell membrane fusion. A soluble, alpha helical, trimeric complex within gp41 composed of N-terminal and C-terminal extraviral segments has been proposed to represent the core of the fusion-active conformation of the HIV-1 envelope. A thermostable subdomain denoted N34(L6)C28 can be formed by the N-34 and C-28 peptides connected by a flexible linker in place of the disulfide-bonded loop region. Three-dimensional structure of N34(L6)C28 reveals that three molecules fold into a six-stranded helical bundle. Three N-terminal helices within the bundle form a central, parallel, trimeric coiled coil, whereas three C-terminal helices pack in the reverse direction into three hydrophobic grooves on the surface of the N-terminal trimer. This thermostable subdomain displays the salient features of the core structure of the isolated gp41 subunit and thus provides a possible target for therapeutics designed selectively to block HIV-1 entry. PMID- 9356445 TI - Expression of hepatocyte nuclear factor 6 in rat liver is sex-dependent and regulated by growth hormone. AB - Growth hormone (GH) binding to its receptor modulates gene transcription by influencing the amount or activity of transcription factors. In the rat, GH exerts sexually dimorphic effects on liver gene transcription through its pattern of secretion which is intermittent in males and continuous in females. The expression of the CYP2C12 gene coding for the female-specific cytochrome P450 2C12 protein is dependent on the continuous exposure to GH. To identify the transcription factor(s) that mediate(s) this sex-dependent GH effect, we studied the interactions of the CYP2C12 promoter with liver nuclear proteins obtained from male and female rats and from hypophysectomized animals treated or not by continuous GH infusion. GH treatment induced the binding of a protein that we identified as hepatocyte nuclear factor (HNF) 6, the prototype of a novel class of homeodomain transcription factors. HNF-6 competed with HNF-3 for binding to the same site in the CYP2C12 promoter. This HNF-6/HNF-3 binding site conveyed both HNF-6- and HNF-3-stimulated transcription of a reporter gene construct in transient cotransfection experiments. Electrophoretic mobility shift assays showed more HNF-6 DNA-binding activity in female than in male liver nuclear extracts. Liver HNF-6 mRNA was barely detectable in the hypophysectomized rats and was restored to normal levels by GH treatment. This work provides an example of a homeodomain-containing transcription factor that is GH-regulated and also reports on the hormonal regulation of HNF-6. PMID- 9356446 TI - Silencer elements controlling the B29 (Igbeta) promoter are neither promoter- nor cell-type-specific. AB - The murine B29 (Igbeta) promoter is B cell specific and contains essential SP1, ETS, OCT, and Ikaros motifs. Flanking 5' DNA sequences inhibit B29 promoter activity, suggesting this region contains silencer elements. Two adjacent 5' DNA segments repress transcription by the murine B29 promoter in a position- and orientation-independent manner, analogous to known silencers. Both these 5' segments also inhibit transcription by several heterologous promoters in B cells, including mb-1, c-fos, and human B29. These 5' segments also inhibit transcription by the c-fos promoter in T cells suggesting they are not B cell specific elements. DNase I footprint analyses show an approximately 70-bp protected region overlapping the boundary between the two negative regulatory DNA segments and corresponding to binding sites for at least two different DNA binding proteins. Within this footprint, two unrelated 30-bp cis-acting DNA motifs (designated TOAD and FROG) function as position- and orientation independent silencers when located directly 5' of the murine B29 promoter. These two silencer motifs act cooperatively to restrict the transcriptional activity of the B29 promoter. Neither of these motifs resembles any known silencers. Mutagenesis of the TOAD and FROG motifs in their respective 5' DNA segments eliminates the silencing activity of these upstream regions, indicating these two motifs as the principal B29 silencer elements within these regions. PMID- 9356447 TI - Peptide nucleic acid-DNA duplexes: long range hole migration from an internally linked anthraquinone. AB - The discovery that peptide nucleic acids (PNA) mimic DNA and RNA by forming complementary duplex structures following Watson-Crick base pairing rules opens fields in biochemistry, diagnostics, and medicine for exploration. Progress requires the development of modified PNA duplexes having unique and well defined properties. We find that anthraquinone groups bound to internal positions of a PNA oligomer intercalate in the PNA-DNA hybrid. Their irradiation with near-UV light leads to electron transfer and oxidative damage at remote GG doublets on the complementary DNA strand. This behavior mimics that observed in related DNA duplexes and provides the first evidence for long range electron (hole) transport in PNA-DNA hybrid. Analysis of the mechanism for electron transport supports hole hopping. PMID- 9356448 TI - Human fatty acid synthase: assembling recombinant halves of the fatty acid synthase subunit protein reconstitutes enzyme activity. AB - Our model of the native fatty acid synthase (FAS) depicts it as a dimer of two identical multifunctional proteins (Mr approximately 272,000) arranged in an antiparallel configuration so that the active Cys-SH of the beta-ketoacyl synthase of one subunit (where the acyl group is attached) is juxtaposed within 2 A of the pantetheinyl-SH of the second subunit (where the malonyl group is bound). This arrangement generates two active centers for fatty acid synthesis and predicts that if we have two appropriate halves of the monomer, we should be able to reconstitute an active fatty acid-synthesizing site. We cloned, expressed, and purified catalytically active thioredoxin (TRX) fusion proteins of the NH2-terminal half of the human FAS subunit protein (TRX-hFAS-dI; residues 1 1,297; Mr approximately 166) and of the C-terminal half (TRX-hFAS-dII-III; residues 1,296-2,504; Mr approximately 155). Adding equivalent amounts of TRX hFAS-dI and TRX-hFAS-dII-III to a reaction mixture containing acetyl-CoA, malonyl CoA, and NADPH resulted in the synthesis of long-chain fatty acids. The rate of synthesis was dependent upon the presence of both recombinant proteins and reached a constant level when they were present in equivalent amounts, indicating that the reconstitution of an active fatty acid-synthesizing site required the presence of every partial activity associated with the subunit protein. Analyses of the product acids revealed myristate to be the most abundant with small amounts of palmitate and stearate, possibly because of the way the fused recombinant proteins interacted with each other so that the thioesterase hydrolyzed the acyl group in its myristoyl state. The successful reconstitution of the human FAS activity from its domain I and domains II and III fully supports our model for the structure-function relationship of FAS in animal tissues. PMID- 9356450 TI - Mosquito transferrin, an acute-phase protein that is up-regulated upon infection. AB - When treated with heat-killed bacterial cells, mosquito cells in culture respond by up-regulating several proteins. Among these is a 66-kDa protein (p66) that is secreted from cells derived from both Aedes aegypti and Aedes albopictus. p66 was degraded by proteolysis and gave a virtually identical pattern of peptide products for each mosquito species. The sequence of one peptide (31 amino acids) was determined and found to have similarity to insect transferrins. By using conserved regions of insect transferrin sequences, degenerate oligonucleotide PCR primers were designed and used to isolate a cDNA clone encoding an A. aegypti transferrin. The encoded protein contained a signal sequence that, when cleaved, would yield a mature protein of 68 kDa. It contained the 31-amino acid peptide, and the 3' end exactly matched a cDNA encoding a polypeptide that is up-regulated when A. aegypti encapsulates filarial worms [Beerntsen, B. T., Severson, D. W. & Christensen, B. M. (1994) Exp. Parasitol. 79, 312-321]. This transferrin, like those of two other insect species, has conserved iron-binding residues in the N terminal lobe but not in the C-terminal lobe, which also has large deletions in the polypeptide chain, compared with transferrins with functional C-terminal lobes. The hypothesis is developed that this transferrin plays a role similar to vertebrate lactoferrin in sequestering iron from invading organisms and that degradation of the structure of the C-terminal lobe might be a mechanism for evading pathogens that elaborate transferrin receptors to tap sequestered iron. PMID- 9356449 TI - TAK, an HIV Tat-associated kinase, is a member of the cyclin-dependent family of protein kinases and is induced by activation of peripheral blood lymphocytes and differentiation of promonocytic cell lines. AB - We have previously identified a cellular protein kinase activity termed TAK that specifically associates with the HIV types 1 and 2 Tat proteins. TAK hyperphosphorylates the carboxyl-terminal domain of the large subunit of RNA polymerase II in vitro in a manner believed to activate transcription [Herrmann, C. H. & Rice, A. P. (1995) J. Virol. 69, 1612-1620]. We show here that the catalytic subunit of TAK is a known human kinase previously named PITALRE, which is a member of the cyclin-dependent family of proteins. We also show that TAK activity is elevated upon activation of peripheral blood mononuclear cells and peripheral blood lymphocytes and upon differentiation of U1 and U937 promonocytic cell lines to macrophages. Therefore, in HIV-infected individuals TAK may be induced in T cells following activation and in macrophages following differentiation, thus contributing to high levels of viral transcription and the escape from latency of transcriptionally silent proviruses. PMID- 9356451 TI - Control of alternative pre-mRNA splicing by distributed pentameric repeats. AB - Multiple copies of the hexamer TGCATG have been shown to regulate fibronectin pre mRNA alternative splicing. GCATG repeats also are clustered near the regulated calcitonin-specific 3' splice site in the rat calcitonin/CGRP gene. Specific mutagenesis of these repeats in calcitonin/CGRP pre-mRNA resulted in the loss of calcitonin-specific splicing, suggesting that the native repeats act to enhance alternative exon inclusion. Mutation of subsets of these elements implies that alternative splicing requires a minimum of two repeats, and that the combination of one intronic and one exonic repeat is necessary for optimal cell-specific splicing. However, multimerized intronic repeats inhibited calcitonin-specific splicing in both the wild-type context and in a transcript lacking endogenous repeats. These results suggest that both the number and distribution of repeats may be important features for the regulation of tissue-specific alternative splicing. Further, RNA containing a single repeat bound cell-specific protein complexes, but tissue-specific differences in protein binding were not detected by using multimerized repeats. Together, these data support a novel model for alternative splicing regulation that requires the cell-specific recognition of multiple, distributed sequence elements. PMID- 9356453 TI - Cholesterol feeding reduces nuclear forms of sterol regulatory element binding proteins in hamster liver. AB - Cholesterol feeding reduces the mRNAs encoding multiple enzymes in the cholesterol biosynthetic pathway and the low density lipoprotein receptor in livers of hamsters. Here we show that cholesterol feeding also reduces the levels of the nuclear NH2-terminal domains of sterol regulatory element binding proteins (SREBPs), which activate transcription of sterol-regulated genes. We show that livers of hamsters, like those of mice and humans, predominantly produce SREBP-2 and the 1c isoform of SREBP-1. Both are produced as membrane-bound precursors that must be proteolyzed to release the transcriptionally active NH2-terminal domains. Diets containing 0.1% to 1.0% cholesterol decreased the amount of nuclear SREBP-1c without affecting the amount of the membrane precursor or its mRNA, suggesting that cholesterol inhibits the proteolytic processing of SREBP-1 in liver as it does in cultured cells. Cholesterol also appeared to reduce the proteolytic processing of SREBP-2. In addition, at high levels of dietary cholesterol the mRNA encoding SREBP-2 declined and the amount of the precursor also fell, suggesting that cholesterol accumulation also may inhibit transcription of the SREBP-2 gene. The high-cholesterol diets reduced the amount of low density lipoprotein receptor mRNA by 30% and produced a more profound 70 90% reduction in mRNAs encoding 3-hydroxy-3-methylglutaryl CoA synthase and reductase. Treatment with lovastatin and Colestipol, which increases hepatic demands for cholesterol, increased the amount of SREBP-2 mRNA as well as the precursor and nuclear forms of the protein. This treatment caused a reciprocal decline in SREBP-1c mRNA and protein. Considered together, these data suggest that SREBPs play important roles in controlling transcription of sterol-regulated genes in liver, as they do in cultured cells. PMID- 9356452 TI - Novel mechanism for carbamoyl-phosphate synthetase: a nucleotide switch for functionally equivalent domains. AB - Carbamoyl-phosphate synthetases (CPSs) utilize two molecules of ATP at two internally duplicated domains, B and C. Domains B and C have recently been shown to be structurally [Thoden, J. B., Holden, H. M., Wesenberg, G., Raushel, F. M. & Rayment, I. (1997) Biochemistry 36, 6305-6316] and functionally [Guy, H. I. & Evans, D. R. (1996) J. Biol. Chem. 271, 13762-13769] equivalent. We have carried out a site-directed mutagenic analysis that is consistent with ATP binding to a palmate motif rather than to a Walker A/B motif in domains B and C. To accommodate our present findings, as well as the other recent findings of structural and functional equivalence, we are proposing a novel mechanism for CPS. In this mechanism utilization of ATP bound to domain C is coupled to carbamoyl-phosphate synthesis at domain B via a nucleotide switch, with the energy of ATP hydrolysis at domain C allowing domain B to cycle between two alternative conformations. PMID- 9356454 TI - Large kinetic isotope effects in enzymatic proton transfer and the role of substrate oscillations. AB - We propose an interpretation of the experimental findings of Klinman and coworkers [Cha, Y., Murray, C. J. & Klinman, J. P. (1989) Science 243, 1325-1330; Grant, K. L. & Klinman, J. P. (1989) Biochemistry 28, 6597-6605; and Bahnson, B. J. & Klinman, J. P. (1995) Methods Enzymol. 249, 373-397], who showed that proton transfer reactions that are catalyzed by bovine serum amine oxidase proceed through tunneling. We show that two different tunneling models are consistent with the experiments. In the first model, the proton tunnels from the ground state. The temperature dependence of the kinetic isotope effect is caused by a thermally excited substrate mode that modulates the barrier, as has been suggested by Borgis and Hynes [Borgis, D. & Hynes, J. T. (1991) J. Chem. Phys. 94, 3619-3628]. In the second model, there is both over-the-barrier transfer and tunneling from excited states. Finally, we propose two experiments that can distinguish between the possible mechanisms. PMID- 9356455 TI - Attenuated T2 relaxation by mutual cancellation of dipole-dipole coupling and chemical shift anisotropy indicates an avenue to NMR structures of very large biological macromolecules in solution. AB - Fast transverse relaxation of 1H, 15N, and 13C by dipole-dipole coupling (DD) and chemical shift anisotropy (CSA) modulated by rotational molecular motions has a dominant impact on the size limit for biomacromolecular structures that can be studied by NMR spectroscopy in solution. Transverse relaxation-optimized spectroscopy (TROSY) is an approach for suppression of transverse relaxation in multidimensional NMR experiments, which is based on constructive use of interference between DD coupling and CSA. For example, a TROSY-type two dimensional 1H,15N-correlation experiment with a uniformly 15N-labeled protein in a DNA complex of molecular mass 17 kDa at a 1H frequency of 750 MHz showed that 15N relaxation during 15N chemical shift evolution and 1HN relaxation during signal acquisition both are significantly reduced by mutual compensation of the DD and CSA interactions. The reduction of the linewidths when compared with a conventional two-dimensional 1H,15N-correlation experiment was 60% and 40%, respectively, and the residual linewidths were 5 Hz for 15N and 15 Hz for 1HN at 4 degrees C. Because the ratio of the DD and CSA relaxation rates is nearly independent of the molecular size, a similar percentagewise reduction of the overall transverse relaxation rates is expected for larger proteins. For a 15N labeled protein of 150 kDa at 750 MHz and 20 degrees C one predicts residual linewidths of 10 Hz for 15N and 45 Hz for 1HN, and for the corresponding uniformly 15N,2H-labeled protein the residual linewidths are predicted to be smaller than 5 Hz and 15 Hz, respectively. The TROSY principle should benefit a variety of multidimensional solution NMR experiments, especially with future use of yet somewhat higher polarizing magnetic fields than are presently available, and thus largely eliminate one of the key factors that limit work with larger molecules. PMID- 9356456 TI - Design of fast enzymes by optimizing interaction potential in active site. AB - The diffusional encounter between substrate and enzyme, and hence catalytic efficiency, can be enhanced by mutating charged residues on the surface of the enzyme. In this paper we present a simple method for screening such mutations. This is based on our earlier result that electrostatic enhancement of the enzyme substrate binding rate constant can be accounted for just by the interaction potential within the active site. Assuming that catalytic and structural integrity is maintained, the catalytic efficiency can be optimized by surface charge mutations which lead to stronger interaction potential within the active site. Application of the screening method on superoxide dismutase shows that only charge mutations close to the active site will have practical effect on the catalytic efficiency. This rationalizes a large number of findings obtained in previous simulation and experimental studies. PMID- 9356457 TI - A method that allows the assembly of kinetochore components onto chromosomes condensed in clarified Xenopus egg extracts. AB - Kinetochores are complex macromolecular structures that link mitotic chromosomes to spindle microtubules. Although a small number of kinetochore components have been identified, including the kinesins CENP-E and XKCM1 as well as cytoplasmic dynein, neither how these and other proteins are organized to produce a kinetochore nor their exact functions within this structure are understood. For this reason, we have developed an assay that allows kinetochore components to assemble onto discrete foci on in vitro-condensed chromosomes. The source of the kinetochore components is a clarified cell extract from Xenopus eggs that can be fractionated or immunodepleted of individual proteins. Kinetochore assembly in these clarified extracts requires preincubating the substrate sperm nuclei in an extract under low ATP conditions. Immunodepletion of XKCM1 from the extracts prevents the localization of kinetochore-associated XKCM1 without affecting the targeting of CENP-E and cytoplasmic dynein or the binding of monomeric tubulin to the kinetochore. Extension of this assay for the analysis of other components should help to dissect the protein-protein interactions involved in kinetochore assembly and function. PMID- 9356458 TI - Hereditary hemochromatosis: effects of C282Y and H63D mutations on association with beta2-microglobulin, intracellular processing, and cell surface expression of the HFE protein in COS-7 cells. AB - Hereditary hemochromatosis (HH) is the most common autosomal recessive disorder known in humans. A candidate gene for HH called HFE has recently been cloned that encodes a novel member of the major histocompatibility complex class I family. Most HH patients are homozygous for a Cys-282-->Tyr (C282Y) mutation in HFE gene, which has been shown to disrupt interaction with beta2-microglobulin; a second mutation, His-63-->Asp (H63D), is enriched in HH patients who are heterozygous for C282Y mutation. The aims of this study were to determine the effects of the C282Y and H63D mutations on the cellular trafficking and degradation of the HFE protein in transfected COS-7 cells. The results indicate that, while the wild type and H63D HFE proteins associate with beta2-microglobulin and are expressed on the cell surface of COS-7 cells, these capabilities are lost by the C282Y HFE protein. We present biochemical and immunofluorescence data that indicate that the C282Y mutant protein: (i) is retained in the endoplasmic reticulum and middle Golgi compartment, (ii) fails to undergo late Golgi processing, and (iii) is subject to accelerated degradation. The block in intracellular transport, accelerated turnover, and failure of the C282Y protein to be presented normally on the cell surface provide a possible basis for impaired function of this mutant protein in HH. PMID- 9356459 TI - The amphiphysin-like protein 1 (ALP1) interacts functionally with the cABL tyrosine kinase and may play a role in cytoskeletal regulation. AB - cABL is a protooncogene, activated in a subset of human leukemias, whose protein product is a nonreceptor tyrosine kinase of unknown function. cABL has a complex structure that includes several domains and motifs found in proteins implicated in signal transduction pathways. An approach to elucidate cABL function is to identify proteins that interact directly with cABL and that may serve as regulators or effectors of its activity. To this end, a protein-interaction screen of a phage expression library was undertaken to identify proteins that interact with specific domains of cABL. An SH3-domain-containing protein has been identified that interacts with sequences in the cABL carboxyl terminus. The cDNA encoding ALP1 (amphiphysin-like protein 1) was isolated from a 16-day mouse embryo. ALP1 has high homology to BIN1, a recently cloned myc-interacting protein, and also shows significant homology to amphiphysin, a neuronal protein cloned from human and chicken. The amino terminus has homology to two yeast proteins, Rvs167 and Rvs161, which are involved in cell entry into stationary phase and cytoskeletal organization. ALP1 binds cABL in vitro and in vivo. Expression of ALP1 results in morphological transformation of NIH 3T3 fibroblasts in a cABL-dependent manner. The properties of ALP1 suggest that it may be involved in possible cytoskeletal functions of the cABL kinase. Additionally, these results provide further evidence for the importance of the cABL carboxyl terminus and its binding proteins in the regulation of cABL function. PMID- 9356460 TI - Hormone-regulatable neoplastic transformation induced by a Jun-estrogen receptor chimera. AB - The v-jun oncogene encodes a nuclear DNA binding protein that functions as a transcription factor and is part of the activator protein 1 complex. Oncogenic transformation by v-jun is thought to be mediated by the aberrant expression of specific target genes. To identify such Jun-regulated genes and to explore the mechanisms by which Jun affects their expression, we have fused the full-length v Jun and an amino-terminally truncated form of v-Jun to the hormone-binding domain of the human estrogen receptor. The two chimeric proteins function as ligand inducible transactivators. Expression of the fusion proteins in chicken embryo fibroblasts causes estrogen-dependent transformation. PMID- 9356461 TI - Bok is a pro-apoptotic Bcl-2 protein with restricted expression in reproductive tissues and heterodimerizes with selective anti-apoptotic Bcl-2 family members. AB - In the intracellular death program, hetero- and homodimerization of different anti- and pro-apoptotic Bcl-2-related proteins are critical in the determination of cell fate. From a rat ovarian fusion cDNA library, we isolated a new pro apoptotic Bcl-2 gene, Bcl-2-related ovarian killer (Bok). Bok had conserved Bcl-2 homology (BH) domains 1, 2, and 3 and a C-terminal transmembrane region present in other Bcl-2 proteins, but lacked the BH4 domain found only in anti-apoptotic Bcl-2 proteins. In the yeast two-hybrid system, Bok interacted strongly with some (Mcl-1, BHRF1, and Bfl-1) but not other (Bcl-2, Bcl-xL, and Bcl-w) anti-apoptotic members. This finding is in direct contrast to the ability of other pro-apoptotic members (Bax, Bak, and Bik) to interact with all of the anti-apoptotic proteins. In addition, negligible interaction was found between Bok and different pro apoptotic members. In mammalian cells, overexpression of Bok induced apoptosis that was blocked by the baculoviral-derived cysteine protease inhibitor P35. Cell killing induced by Bok was also suppressed following coexpression with Mcl-1 and BHRF1 but not with Bcl-2, further indicating that Bok heterodimerized only with selective anti-apoptotic Bcl-2 proteins. Northern blot analysis indicated that Bok was highly expressed in the ovary, testis and uterus. In situ hybridization analysis localized Bok mRNA in granulosa cells, the cell type that underwent apoptosis during follicle atresia. Identification of Bok as a new pro-apoptotic Bcl-2 protein with restricted tissue distribution and heterodimerization properties could facilitate elucidation of apoptosis mechanisms in reproductive tissues undergoing hormone-regulated cyclic cell turnover. PMID- 9356462 TI - Nonmuscle myosin II-B is required for normal development of the mouse heart. AB - We used targeted gene disruption in mice to ablate nonmuscle myosin heavy chain B (NMHC-B), one of the two isoforms of nonmuscle myosin II present in all vertebrate cells. Approximately 65% of the NMHC-B-/- embryos died prior to birth, and those that were born suffered from congestive heart failure and died during the first day. No abnormalities were detected in NMHC-B+/- mice. The absence of NMHC-B resulted in a significant increase in the transverse diameters of the cardiac myocytes from 7.8 +/- 1.8 micron (right ventricle) and 7.8 +/- 1.3 micron (left ventricle) in NMHC-B+/+ and B+/- mice to 14.7 +/- 1.1 micron and 13.8 +/- 2.3 micron, respectively, in NMHC-B-/- mice (in both cases, P < 0.001). The increase in size of the cardiac myocytes was seen as early as embryonic day 12.5 (4.5 +/- 0.2 micron for NMHC-B+/+ and B+/- vs. 7. 2 +/- 0.6 micron for NMHC-B-/- mice (P < 0.01)). Six of seven NMHC-B-/- newborn mice analyzed by serial sectioning also showed structural cardiac defects, including a ventricular septal defect, an aortic root that either straddled the defect or originated from the right ventricle, and muscular obstruction to right ventricular outflow. Some of the hearts of NMHC-B-/- mice showed evidence for up-regulation of NMHC-A protein. These studies suggest that nonmuscle myosin II-B is required for normal cardiac myocyte development and that its absence results in structural defects resembling, in part, two common human congenital heart diseases, tetralogy of Fallot and double outlet right ventricle. PMID- 9356463 TI - Dystrobrevin and dystrophin: an interaction through coiled-coil motifs. AB - Dystrobrevin, a dystrophin-related and -associated protein, has been proposed to be important in the formation and maintenance of the neuromuscular junction. Dystrobrevin coprecipitates with both the acetylcholine receptor complex as well as the dystrophin glycoprotein complex. Although the nature of dystrobrevin's association with the dystrophin glycoprotein complex remains unclear, it is known that dystrobrevin binds directly to the syntrophins, a heterologous group of dystrophin-associated proteins. Using the yeast two-hybrid system to identify protein-protein interactions, we present evidence for the heterodimerization of dystrobrevin directly with dystrophin. The C terminus of dystrobrevin binds specifically to the C terminus of dystrophin. We further refined this site of interaction to these proteins' homologous coiled-coil motifs that flank their respective syntrophin-binding sites. We also show that the interaction between the dystrobrevin and dystrophin coiled-coil domains is specific and is not due to a nonspecific coiled-coil domain interaction. From the accumulated evidence of protein-protein interactions presented here and elsewhere, we propose a partially revised model of the organization of the dystrophin-associated glycoprotein complex. PMID- 9356464 TI - Grb2-associated binder-1 mediates phosphatidylinositol 3-kinase activation and the promotion of cell survival by nerve growth factor. AB - Nerve growth factor (NGF) prevents apoptosis through stimulation of the TrkA receptor protein tyrosine kinase. The downstream activation of phosphatidylinositol 3-kinase (PI 3-kinase) is essential for the inhibition of apoptosis, although this enzyme does not bind to and is not directly activated by TrkA. We have found that the addition of NGF to PC-12 cells resulted in the phosphorylation of the Grb2-associated binder-1 (Gab1) docking protein and induced the association of several SH2 domain-containing proteins, including PI 3 kinase. A substantial fraction of the total cellular PI 3-kinase activity was associated with Gab1. PC-12 cells that overexpressed Gab1 show a decreased requirement for the amount of NGF necessary to inhibit apoptosis. The expression of a Gab1 mutant that lacked the binding sites for PI 3-kinase enhanced apoptosis and diminished the protective effect of NGF. Hence, Gab1 has a major role in connecting TrkA with PI 3-kinase activation and for the promotion of cell survival by NGF. PMID- 9356465 TI - Prominin, a novel microvilli-specific polytopic membrane protein of the apical surface of epithelial cells, is targeted to plasmalemmal protrusions of non epithelial cells. AB - Using a new mAb raised against the mouse neuroepithelium, we have identified and cDNA-cloned prominin, an 858-amino acid-containing, 115-kDa glycoprotein. Prominin is a novel plasma membrane protein with an N-terminal extracellular domain, five transmembrane segments flanking two short cytoplasmic loops and two large glycosylated extracellular domains, and a cytoplasmic C-terminal domain. DNA sequences from Caenorhabditis elegans predict the existence of a protein with the same features, suggesting that prominin is conserved between vertebrates and invertebrates. Prominin is found not only in the neuroepithelium but also in various other epithelia of the mouse embryo. In the adult mouse, prominin has been detected in the brain ependymal layer, and in kidney tubules. In these epithelia, prominin is specific to the apical surface, where it is selectively associated with microvilli and microvilli-related structures. Remarkably, upon expression in CHO cells, prominin is preferentially localized to plasma membrane protrusions such as filopodia, lamellipodia, and microspikes. These observations imply that prominin contains information to be targeted to, and/or retained in, plasma membrane protrusions rather than the planar cell surface. Moreover, our results show that the mechanisms underlying targeting of membrane proteins to microvilli of epithelial cells and to plasma membrane protrusions of non epithelial cells are highly related. PMID- 9356467 TI - alpha-tocopherol transfer protein stimulates the secretion of alpha-tocopherol from a cultured liver cell line through a brefeldin A-insensitive pathway. AB - Vitamin E (alpha-tocopherol) is a fat-soluble antioxidant that is transported by plasma lipoproteins in the body. alpha-Tocopherol taken up by the liver with lipoprotein is thought to be resecreted into the plasma in very low density lipoprotein (VLDL). alpha-Tocopherol transfer protein (alphaTTP), which was recently identified as a product of the causative gene for familial isolated vitamin E deficiency, is a cytosolic liver protein and plays an important role in the efficient recycling of plasma vitamin E. To throw light on the mechanism of alphaTTP-mediated alpha-tocopherol transfer in the liver cell, we devised an assay system using the hepatoma cell line McARH7777. Using this system, we found that the secretion of alpha-tocopherol was more efficient in cells expressing alphaTTP than in matched cells lacking alphaTTP. Brefeldin A, which effectively inhibits VLDL secretion by disrupting the Golgi apparatus, had no effect on alpha tocopherol secretion, indicating that alphaTTP-mediated alpha-tocopherol secretion is not coupled to VLDL secretion. Among other agents tested, only 25 hydroxycholesterol, a modulator of cholesterol metabolism, inhibited alpha tocopherol secretion. This inhibition is most likely mediated by oxysterol binding protein. These results suggest that alphaTTP present in the liver cytosol functions to stimulate secretion of cellular alpha-tocopherol into the extracellular medium and that the reaction utilizes a novel non-Golgi-mediated pathway that may be linked to cellular cholesterol metabolism and/or transport. PMID- 9356468 TI - Is sexual selection and species recognition a continuum? Mating behavior of the stalk-eyed fly Drosophila heteroneura. AB - If behavioral isolation between species can evolve as a consequence of sexual selection within a species, then traits that are both sexually selected and used as a criterion of species recognition by females should be identifiable. The broad male head of the Hawaiian picture-winged fly Drosophila heteroneura is a novel sexual dimorphism that may be sexually selected and involved in behavioral isolation from D. silvestris. We found that males with broad heads are more successful in sexual selection, both through female mate choice and through aggressive interactions. However, female D. heteroneura do not discriminate against hybrids on the basis of their head width. Thus, this novel trait is sexually selected but is not a major contributor to species recognition. Our methods should be applicable to other species in which behavioral isolation is a factor. PMID- 9356466 TI - MAD2 associates with the cyclosome/anaphase-promoting complex and inhibits its activity. AB - Cell cycle progression is monitored by checkpoint mechanisms that ensure faithful duplication and accurate segregation of the genome. Defects in spindle assembly or spindle-kinetochore attachment activate the mitotic checkpoint. Once activated, this checkpoint arrests cells prior to the metaphase-anaphase transition with unsegregated chromosomes, stable cyclin B, and elevated M phase promoting factor activity. However, the mechanisms underlying this process remain obscure. Here we report that upon activation of the mitotic checkpoint, MAD2, an essential component of the mitotic checkpoint, associates with the cyclin B ubiquitin ligase, known as the cyclosome or anaphase-promoting complex. Moreover, purified MAD2 causes a metaphase arrest in cycling Xenopus laevis egg extracts and prevents cyclin B proteolysis by blocking its ubiquitination, indicating that MAD2 functions as an inhibitor of the cyclosome. Thus, MAD2 links the mitotic checkpoint pathway to the cyclin B destruction machinery which is critical in controlling the metaphase-anaphase transition. PMID- 9356469 TI - Expression of multiple insulin and insulin-like growth factor receptor genes in salmon gill cartilage. AB - In mammals, one of the major actions of insulin-like growth factor I (IGF-I) is to increase skeletal growth by stimulating new cartilage formation. IGF-I stimulates chondrocytes in vitro to synthesize new cartilage matrix, measured by enhanced uptake of 35S-sulfate, but the addition of insulin does not produce a similar effect except when added at high concentrations. However, recent studies have shown that, in teleosts, both insulin and IGF-I are potent activators of 35S sulfate uptake in gill cartilage. To further characterize the growth-promoting activities of these hormones in fish, we have used reverse transcriptase-linked PCR to analyze the expression of insulin receptor family genes in salmon gill cartilage. Partial cDNA sequences encoding the tyrosine kinase domains from six distinct members of the IR gene family were obtained, and sequence comparisons revealed that four of the cDNAs encoded amino acid sequences that were highly homologous to human IR whereas the encoded sequences from two of the cDNAs were more similar to the human type I IGF receptor (IGF-R). Furthermore, a comparative reverse transcriptase-linked PCR assay revealed that the four putative IR mRNAs expressed in toto in gill cartilage were 56% of that found in liver whereas the expressed amount of the two IGF-R mRNAs was 9-fold higher compared with liver. These results suggest that the chondrogenic actions of insulin and IGF-I in fish are mediated by the ligands binding to their cognate receptors. However, further studies will be required to characterize the binding properties and relative contribution of the individual IR and IGF-R genes. PMID- 9356470 TI - Strong balancing selection at HLA loci: evidence from segregation in South Amerindian families. AB - The genotypic proportions for major histocompatibility complex loci, HLA-A and HLA-B, of progeny in families in 23 South Amerindian tribes in which segregation for homozygotes and heterozygotes could occur are examined. Overall, there is a large deficiency of homozygotes compared with Mendelian expectations (for HLA-A, 114 observed and 155.50 expected and for HLA-B 110 observed and 144.75 expected), consistent with strong balancing selection favoring heterozygotes. There is no evidence that these deficiencies were associated with particular alleles or with the age of the individuals sampled. When these families were divided into four mating types, there was strong selection against homozygotes, averaging 0.462 for three of the mating types over the two loci. For the other mating type in which the female parent is homozygous and shares one allele with the heterozygous male parent, there was no evidence of selection against homozygotes. A theoretical model incorporating these findings surprisingly does not result in a stable polymorphism for two alleles but does result in an excess of heterozygotes and a minimum fitness at intermediate allele frequencies. However, for more than two alleles, balancing selection does occur and the model approaches the qualities of the symmetrical heterozygote advantage model as the number of alleles increases. PMID- 9356471 TI - Double muscling in cattle due to mutations in the myostatin gene. AB - Myostatin (GDF-8) is a member of the transforming growth factor beta superfamily of secreted growth and differentiation factors that is essential for proper regulation of skeletal muscle mass in mice. Here we report the myostatin sequences of nine other vertebrate species and the identification of mutations in the coding sequence of bovine myostatin in two breeds of double-muscled cattle, Belgian Blue and Piedmontese, which are known to have an increase in muscle mass relative to conventional cattle. The Belgian Blue myostatin sequence contains an 11-nucleotide deletion in the third exon which causes a frameshift that eliminates virtually all of the mature, active region of the molecule. The Piedmontese myostatin sequence contains a missense mutation in exon 3, resulting in a substitution of tyrosine for an invariant cysteine in the mature region of the protein. The similarity in phenotypes of double-muscled cattle and myostatin null mice suggests that myostatin performs the same biological function in these two species and is a potentially useful target for genetic manipulation in other farm animals. PMID- 9356472 TI - Transgenic rats reveal functional conservation of regulatory controls between the Fugu isotocin and rat oxytocin genes. AB - We have asked whether comparative genome analysis and rat transgenesis can be used to identify functional regulatory domains in the gene locus encoding the hypothalamic neuropeptides oxytocin (OT) and vasopressin. Isotocin (IT) and vasotocin (VT) are the teleost homologues of these genes. A contiguous stretch of 46 kb spanning the Fugu IT-VT locus has been sequenced, and nine putative genes were found. Unlike the OT and vasopressin genes, which are closely linked in the mammalian genome in a tail-to-tail orientation, Fugu IT and VT genes are linked head to tail and are separated by five genes. When a cosmid containing the Fugu IT-VT locus was introduced into the rat genome, we found that the Fugu IT gene was specifically expressed in rat hypothalamic oxytocinergic neurons and mimicked the response of the endogenous OT gene to an osmotic stimulus. These data show that cis-acting elements and trans-acting factors mediating the cell-specific and physiological regulation of the OT and IT genes are conserved between mammals and fish. The combination of Fugu genome analysis and transgenesis in a mammal is a powerful tool for identifying and analyzing conserved vertebrate regulatory elements. PMID- 9356473 TI - Intron self-complementarity enforces exon inclusion in a yeast pre-mRNA. AB - Skipping of internal exons during removal of introns from pre-mRNA must be avoided for proper expression of most eukaryotic genes. Despite significant understanding of the mechanics of intron removal, mechanisms that ensure inclusion of internal exons in multi-intron pre-mRNAs remain mysterious. Using a natural two-intron yeast gene, we have identified distinct RNA-RNA complementarities within each intron that prevent exon skipping and ensure inclusion of internal exons. We show that these complementarities are positioned to act as intron identity elements, bringing together only the appropriate 5' splice sites and branchpoints. Destroying either intron self-complementarity allows exon skipping to occur, and restoring the complementarity using compensatory mutations rescues exon inclusion, indicating that the elements act through formation of RNA secondary structure. Introducing new pairing potential between regions near the 5' splice site of intron 1 and the branchpoint of intron 2 dramatically enhances exon skipping. Similar elements identified in single intron yeast genes contribute to splicing efficiency. Our results illustrate how intron secondary structure serves to coordinate splice site pairing and enforce exon inclusion. We suggest that similar elements in vertebrate genes could assist in the splicing of very large introns and in the evolution of alternative splicing. PMID- 9356474 TI - Cells that register logical relationships among proteins. AB - Two-hybrid methods have augmented the classical genetic techniques biologists use to assign function to genes. Here, we describe construction of a two-bait interaction trap that uses yeast cells to register more complex protein relationships than those detected in existing two-hybrid systems. We show that such cells can identify bridge or connecting proteins and peptide aptamers that discriminate between closely related allelic variants. The protein relationships detected by these cells are analogous to classical genetic relationships, but lend themselves to systematic application to the products of entire genomes and combinatorial libraries. We show that, by performing logical operations on the phenotypic outputs of these complex cells and existing two-hybrid cells, we can make inferences about the topology and order of protein interactions. Finally, we show that cells that register such relationships can perform logical operations on protein inputs. Thus these cells will be useful for analysis of gene and allele function, and may also define a path for construction of biological computational devices. PMID- 9356475 TI - Growth suppression of glioma cells by PTEN requires a functional phosphatase catalytic domain. AB - Deletions of all or part of chromosome 10 are the most common genetic alterations in high-grade gliomas. The PTEN gene (also called MMAC1 and TEP1) maps to chromosome region 10q23 and has been implicated as a target of alteration in gliomas and also in other cancers such as those of the breast, prostate, and kidney. Here we sought to provide a functional test of its candidacy as a growth suppressor in glioma cells. We used a combination of Northern blot analysis, protein truncation assays, and sequence analysis to determine the types and frequency of PTEN mutations in glioma cell lines so that we could define appropriate recipients to assess the growth suppressive function of PTEN by gene transfer. Introduction of wild-type PTEN into glioma cells containing endogenous mutant alleles caused growth suppression, but was without effect in cells containing endogenous wild-type PTEN. The ectopic expression of PTEN alleles, which carried mutations found in primary tumors and have been shown or are expected to inactivate its phosphatase activity, caused little growth suppression. These data strongly suggest that PTEN is a protein phosphatase that exhibits functional and specific growth-suppressing activity. PMID- 9356477 TI - Schizosaccharomyces pombe cdc20+ encodes DNA polymerase epsilon and is required for chromosomal replication but not for the S phase checkpoint. AB - In fission yeast both DNA polymerase alpha (pol alpha) and delta (pol delta) are required for DNA chromosomal replication. Here we demonstrate that Schizosaccharomyces pombe cdc20+ encodes the catalytic subunit of DNA polymerase epsilon (pol epsilon) and that this enzyme is also required for DNA replication. Following a shift to the restrictive temperature, cdc20 temperature-sensitive mutant cells block at the onset of DNA replication, suggesting that cdc20+ is required early in S phase very near to the initiation step. In the budding yeast Saccharomyces cerevisiae, it has been reported that in addition to its proposed role in chromosomal replication, DNA pol epsilon (encoded by POL2) also functions directly as an S phase checkpoint sensor [Navas, T. A., Zhou, Z. & Elledge, S. J. (1995) Cell 80, 29-39]. We have investigated whether cdc20+ is required for the checkpoint control operating in fission yeast, and our data indicate that pol epsilon does not have a role as a checkpoint sensor coordinating S phase with mitosis. In contrast, germinating spores disrupted for the gene encoding pol alpha rapidly enter mitosis in the absence of DNA synthesis, suggesting that in the absence of pol alpha, normal coordination between S phase and mitosis is lost. We propose that the checkpoint signal operating in S phase depends on assembly of the replication initiation complex, and that this signal is generated prior to the elongation stage of DNA synthesis. PMID- 9356476 TI - Biological characterization of Drosophila Rapgap1, a GTPase activating protein for Rap1. AB - The activity of Ras family proteins is modulated in vivo by the function of GTPase activating proteins, which increase their intrinsic rate of GTP hydrolysis. We have isolated cDNAs encoding a GAP for the Drosophila Rap1 GTPase. Drosophila Rapgap1 encodes an 850-amino acid protein with a central region that displays substantial sequence similarity to human RapGAP. This domain, when expressed in Escherichia coli, potently stimulates Rap1 GTPase activity in vitro. Unlike Rap1, which is ubiquitously expressed, Rapgap1 expression is highly restricted. Rapgap1 is expressed at high levels in the developing photoreceptor cells and in the optic lobe. Rapgap1 mRNA is also localized in the pole plasm in an oskar-dependent manner. Although mutations that completely abolish Rapgap1 function display no obvious phenotypic abnormalities, overexpression of Rapgap1 induces a rough eye phenotype that is exacerbated by reducing Rap1 gene dosage. Thus, Rapgap1 can function as a negative regulator of Rap1-mediated signaling in vivo. PMID- 9356478 TI - The absence of effect of gid or mioC transcription on the initiation of chromosomal replication in Escherichia coli. AB - Despite the widely accepted view that transcription of gid and mioC is required for efficient initiation of cloned oriC, we show that these transcriptions have very little effect on initiation of chromosome replication at wild-type chromosomal oriC. Furthermore, neither gid nor mioC transcription is required in cells deficient in the histone-like proteins Fis or IHF. However, oriC that is sufficiently impaired for initiation by deletion of DnaA box R4 requires transcription of at least one of these genes. We conclude that transcription of mioC and especially gid is needed to activate oriC only under suboptimal conditions. We suggest that either the rifampicin-sensitive step of initiation is some other transcription occurring from promoter(s) within oriC, or the original inference of transcriptional activation derived from the rifampicin experiments is incorrect. PMID- 9356479 TI - The prion model for [URE3] of yeast: spontaneous generation and requirements for propagation. AB - The genetic properties of the non-Mendelian element, [URE3], suggest that it is a prion (infectious protein) form of Ure2p, a mediator of nitrogen regulation in Saccharomyces cerevisiae. Into a ure2Delta strain (necessarily lacking [URE3]), we introduced a plasmid overproducing Ure2p. This induced the frequent "spontaneous generation" of [URE3], with properties identical to the original [URE3]. Altering the translational frame only in the prion-inducing domain of URE2 shows that it is Ure2 protein (and not URE2 RNA) that induces appearance of [URE3]. The proteinase K-resistance of Ure2p is unique to [URE3] strains and is not seen in nitrogen regulation of normal strains. The prion-inducing domain of Ure2p (residues 1-65) can propagate [URE3] in the absence of the C-terminal part of the molecule. In contrast, the C-terminal part of Ure2p cannot be converted to the prion (inactive) form without the prion-inducing domain covalently attached. These experiments support the prion model for [URE3] and extend our understanding of its propagation. PMID- 9356480 TI - Multiple imprinted sense and antisense transcripts, differential methylation and tandem repeats in a putative imprinting control region upstream of mouse Igf2. AB - The mouse insulin-like growth factor 2 (Igf2) locus is a complex genomic region that produces multiple transcripts from alternative promoters. Expression at this locus is regulated by parental imprinting. However, despite the existence of putative imprinting control elements in the Igf2 upstream region, imprinted transcriptional repression is abolished by null mutations at the linked H19 locus. To clarify the extent to which the Igf2 upstream region contains autonomous imprinting control elements we have performed functional and comparative analyses of the region in the mouse and human. Here we report the existence of multiple, overlapping imprinted (maternally repressed) sense and antisense transcripts that are associated with a tandem repeat in the mouse Igf2 upstream region. Regions flanking the repeat exhibit tissue-specific parental allelic methylation patterns, suggesting the existence of tissue-specific control elements in the upstream region. Studies in H19 null mice indicate that both parental allelic methylation and monoallelic expression of the upstream transcripts depends on an intact H19 gene acting in cis. The homologous region in human IGF2 is structurally conserved, with the significant exception that it does not contain a tandem repeat. Our results support the proposal that tandem repeats act to target methylation to imprinted genetic loci. PMID- 9356481 TI - Genetic interactions with Rap1 and Ras1 reveal a second function for the fat facets deubiquitinating enzyme in Drosophila eye development. AB - The Drosophila fat facets gene encodes a deubiquitinating enzyme that regulates a cell communication pathway essential very early in eye development, prior to facet assembly, to limit the number of photoreceptor cells in each facet of the compound eye to eight. The Fat facets protein facilitates the production of a signal in cells outside the developing facets that inhibits neural development of particular facet precursor cells. Novel gain-of-function mutations in the Drosophila Rap1 and Ras1 genes are described herein that interact genetically with fat facets mutations. Analysis of these genetic interactions reveals that Fat facets has an additional function later in eye development involving Rap1 and Ras1 proteins. Moreover, the results suggest that undifferentiated cells outside the facet continue to influence facet assembly later in eye development. PMID- 9356482 TI - CDC45 is required in conjunction with CDC7/DBF4 to trigger the initiation of DNA replication. AB - The initiation of DNA replication in Saccharomyces cerevisiae requires the protein product of the CDC45 gene. We report that although Cdc45p is present at essentially constant levels throughout the cell cycle, it completes its initiation function in late G1, after START and prior to DNA synthesis. Shortly after mitosis, cells prepare for initiation by assembling prereplicative complexes at their replication origins. These complexes are then triggered at the onset of S phase to commence DNA replication. Cells defective for CDC45 are incapable of activating the complexes to initiate DNA replication. In addition, Cdc45p and Cdc7p/Dbf4p, a kinase implicated in the G1/S phase transition, are dependent on one another for function. These data indicate that CDC45 functions in late G1 phase in concert with CDC7/DBF4 to trigger initiation at replication origins after the assembly of the prereplicative complexes. PMID- 9356483 TI - Parasite-mediated nuclear factor kappaB regulation in lymphoproliferation caused by Theileria parva infection. AB - Infection of cattle with the protozoan Theileria parva results in uncontrolled T lymphocyte proliferation resulting in lesions resembling multicentric lymphoma. Parasitized cells exhibit autocrine growth characterized by persistent translocation of the transcriptional regulatory factor nuclear factor kappaB (NFkappaB) to the nucleus and consequent enhanced expression of interleukin 2 and the interleukin 2 receptor. How T. parva induces persistent NFkappaB activation, required for T cell activation and proliferation, is unknown. We hypothesized that the parasite induces degradation of the IkappaB molecules which normally sequester NFkappaB in the cytoplasm and that continuous degradation requires viable parasites. Using T. parva-infected T cells, we showed that the parasite mediates continuous phosphorylation and proteolysis of IkappaBalpha. However, IkappaBalpha reaccumulated to high levels in parasitized cells, which indicated that T. parva did not alter the normal NFkappaB-mediated positive feedback loop which restores cytoplasmic IkappaBalpha. In contrast, T. parva mediated continuous degradation of IkappaBbeta resulting in persistently low cytoplasmic IkappaBbeta levels. Normal IkappaBbeta levels were only restored following T. parva killing, indicating that viable parasites are required for IkappaBbeta degradation. Treatment of T. parva-infected cells with pyrrolidine dithiocarbamate, a metal chelator, blocked both IkappaB degradation and consequent enhanced expression of NFkappaB dependent genes. However treatment using the antioxidant N-acetylcysteine had no effect on either IkappaB levels or NFkappaB activation, indicating that the parasite subverts the normal IkappaB regulatory pathway downstream of the requirement for reactive oxygen intermediates. Identification of the critical points regulated by T. parva may provide new approaches for disease control as well as increase our understanding of normal T cell function. PMID- 9356485 TI - ER-27319, an acridone-related compound, inhibits release of antigen-induced allergic mediators from mast cells by selective inhibition of fcepsilon receptor I-mediated activation of Syk. AB - Engagement of the mast cell high-affinity receptor for immunoglobulin E (IgE), FcepsilonRI, induces tyrosine phosphorylation of Syk, a non-receptor tyrosine kinase, that has been demonstrated as critical for degranulation. Herein we describe a synthetic compound, ER-27319, as a potent and selective inhibitor of antigen or anti-IgE-mediated degranulation of rodent and human mast cells. ER 27319 affected neither Lyn kinase activity nor the antigen-induced phosphorylation of the FcepsilonRI but did effectively inhibit the tyrosine phosphorylation of Syk and thus its activity. As a consequence, tyrosine phosphorylation of phospholipase C-gamma1, generation of inositol phosphates, release of arachidonic acid, and secretion of histamine and tumor necrosis factor alpha were also inhibited. ER-27319 did not inhibit the anti-CD3-induced tyrosine phosphorylation of phospholipase C-gamma1 in Jurkat T cells, demonstrating a specificity for Syk-induced signals. In contrast the tyrosine phosphorylation and activation of Syk, induced by in vitro incubation with the phosphorylated immunoreceptor tyrosine-based activation motif (ITAM) of FcepsilonRI gamma subunit or by antigen activation of RBL-2H3 cells, was specifically inhibited by ER-27319. However, when ER-27319 was added to immunoprecipitated Syk, derived from activated cells, no effect was seen on Syk activity. ER-27319 did not inhibit the tyrosine phosphorylation of Syk induced by activation in the presence of Igbeta ITAM or the anti-IgM-induced phosphorylation of Syk in human peripheral B cells. Therefore, ER-27319 selectively interferes with the FcepsilonRI gamma phospho-ITAM activation of Syk in vitro and in intact cells. These results confirm the importance of Syk in FcepsilonRI-mediated responses in mast cells and demonstrate the mast cell selectivity and therapeutic potential of ER-27319 in the treatment of allergic disease. PMID- 9356484 TI - Therapeutic passive vaccination against chronic Lyme disease in mice. AB - Passive and active immunization against outer surface protein A (OspA) has been successful in protecting laboratory animals against subsequent infection with Borrelia burgdorferi. Antibodies (Abs) to OspA convey full protection, but only when they are present at the time of infection. Abs inactivate spirochetes within the tick and block their transmission to mammals, but do not affect established infection because of the loss of OspA in the vertebrate host. Our initial finding that the presence of high serum titers of anti-OspC Abs (5 to 10 microg/ml) correlates with spontaneous resolution of disease and infection in experimentally challenged immunocompetent mice suggested that therapeutic vaccination with OspC may be feasible. We now show that polyclonal and monospecific mouse immune sera to recombinant OspC, but not to OspA, of B. burgdorferi resolve chronic arthritis and carditis and clear disseminated spirochetes in experimentally infected C.B. 17 severe combined immunodeficient mice in a dose-dependent manner. This was verified by macroscopical and microscopical examination of affected tissues and recultivation of spirochetes from ear biopsies. Complete resolution of disease and infection was achieved, independent of whether OspC-specific immune sera (10 microg OspC-specific Abs) were repeatedly given (4x in 3- to 4-day intervals) before the onset (day 10 postinfection) or at the time of fully established arthritis and carditis (days 19 or 60 postinfection). The results indicate that in mice spirochetes constitutively express OspC and are readily susceptible to protective OspC-specific Abs throughout the infection. Thus, an OspC-based vaccine appears to be a candidate for therapy of Lyme disease. PMID- 9356486 TI - A retro-inverso peptide corresponding to the GH loop of foot-and-mouth disease virus elicits high levels of long-lasting protective neutralizing antibodies. AB - Peptides corresponding to the immunodominant loop located at residues 135-158 on capsid protein VP1 of foot-and-mouth disease virus (FMDV) generally elicit high levels of anti-peptide and virus-neutralizing antibodies. In some instances, however, the level of neutralizing antibodies is low or even negligible, even though the level of anti-peptide antibodies is high. We have shown previously that the antigenic activity of peptide 141-159 of VP1 of a variant of serotype A can be mimicked by a retro-inverso (all-D retro or retroenantio) peptide analogue. This retro-inverso analogue induced greater and longer-lasting antibody titers than did the corresponding L-peptide. We now show that a single inoculation of the retro-inverso analogue elicits high levels of neutralizing antibodies that persist longer than those induced against the corresponding L peptide and confer substantial protection in guinea pigs challenged with the cognate virus. In view of the high stability to proteases of retro-inverso peptide analogues and their enhanced immunogenicity, these results have practical relevance in designing potential peptide vaccines. PMID- 9356487 TI - Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin. AB - Dendritic cells (DC) have been thought to represent a family of closely related cells with similar functions and developmental pathways. The best-characterized precursors are the epidermal Langerhans cells, which migrate to lymphoid organs and become activated DC in response to inflammatory stimuli. Here, we demonstrate that a large subset of DC in the T cell-dependent areas of human lymphoid organs are nonactivated cells and belong to a separate lineage that can be identified by high levels of the interleukin 3 receptor alpha chain (IL-3Ralphahi). The CD34+IL 3Ralphahi DC progenitors are of myeloid origin and are distinct from those that give rise to Langerhans cells in vitro. The IL-3Ralphahi DC furthermore appear to migrate to lymphoid organs independently of inflammatory stimuli or foreign antigens. Thus, DC are heterogeneous with regard to function and ontogeny. PMID- 9356488 TI - T cell functions in granulocyte/macrophage colony-stimulating factor deficient mice. AB - Immunological functions were analyzed in mice lacking granulocyte/macrophage colony-stimulating factor (GM-CSF). The response of splenic T cells to allo antigens, anti-mouse CD3 mAb, interleukin 2 (IL-2), or concanavalin A was comparable in GM-CSF +/+ and GM-CSF -/- mice. To investigate the responses of CD8(+) and CD4+ T cells against exogenous antigens, mice were immunized with ovalbumin peptide or with keyhole limpet hemocyanin (KLH). Cytotoxic CD8+ T cells with specificity for ovalbumin peptide could not be induced in GM-CSF -/- mice. After immunization with KLH, there was a delay in IgG generation, particularly IgG2a, in GM-CSF -/- mice. Purified CD4+ T cells from GM-CSF -/- mice immunized with KLH showed impaired proliferative responses and produced low amounts of interferon-gamma (IFN-gamma) and IL-4 when KLH-pulsed B cells or spleen cells were used as antigen presenting cells (APC). When enriched dendritic cells (DC) were used as APC, CD4+ T cells from GM-CSF -/- mice proliferated as well as those from GM-CSF +/+ mice and produced high amounts of IFN-gamma and IL-4. To analyze the rescue effect of DC on CD4(+) T cells, supernatants from (i) CD4(+) T cells cultured with KLH-pulsed DC or (ii) DC cultured with recombinant GM-CSF were transferred to cultures of CD4(+) T cells and KLH-pulsed spleen cells from GM-CSF -/- mice. Supernatants from both DC sources contained a factor or factors that restored proliferative responses and IFN-gamma production of CD4(+) T cells from GM-CSF -/- mice. PMID- 9356489 TI - A very limited number of keywords (main patterns) describes all sequences of the human variable heavy (VH) and kappa (Vkappa) domains. AB - Sequences of the variable heavy (VH) and kappa (Vkappa) domains of Ig structures were divided into 21 fragments that correspond to strands, loops, or parts of these structural units of the variable domains. Amino acid sequences of fragments (termed "words") were collected from the 1,172 human heavy and 668 human kappa chains available in the Kabat database. Statistical analysis of words of 17 fragments was performed (fragments that comprise the complementary determining regions' fragments will not be discussed in this paper). The number of different words (those with different residues in at least one position) ranged, for various fragments, from 11 to 75 in the kappa chains, and from 23 to 189 in the heavy chains. The main result of this study is that very few keywords, or main patterns of words, were necessary to describe over 90% of the sequences (no more than two keywords per fragment in the kappa and no more than five per fragment in the heavy chains). No identical keywords were found for different fragments of the variable domains. Keywords of aligned fragments of the VH and Vkappa domains were different in all but two instances. Thus, knowing the keywords, one can determine whether any given small part of a sequence belongs to a heavy or kappa chain and predict its precise localization in the sequence. In addition, by using all of the keywords obtained through analysis of the Kabat database, it was possible to describe completely the sequences of the human VH and Vkappa germ line segments. PMID- 9356490 TI - Effects of in vivo adventitial expression of recombinant endothelial nitric oxide synthase gene in cerebral arteries. AB - Nitric oxide (NO), synthesized from L-arginine by NO synthases (NOS), plays an essential role in the regulation of cerebrovascular tone. Adenoviral vectors have been widely used to transfer recombinant genes to different vascular beds. To determine whether the recombinant endothelial NOS (eNOS) gene can be delivered in vivo to the adventitia of cerebral arteries and functionally expressed, a replication-incompetent adenoviral vector encoding eNOS gene (AdCMVNOS) or beta galactosidase reporter gene (AdCMVLacZ) was injected into canine cerebrospinal fluid (CSF) via the cisterna magna (final viral titer in CSF, 10(9) pfu/ml). Adventitial transgene expression was demonstrated 24 h later by beta galactosidase histochemistry and quantification, eNOS immunohistochemistry, and Western blot analysis of recombinant eNOS. Electron microscopy immunogold labeling indicated that recombinant eNOS protein was expressed in adventitial fibroblasts. In AdCMVNOS-transduced arteries, basal cGMP production and bradykinin-induced relaxations were significantly augmented when compared with AdCMVLacZ-transduced vessels (P < 0.05). The increased receptor-mediated relaxations and cGMP production were inhibited by eNOS inhibitors. In addition, the increase in cGMP production was reversed in the absence of calcium, suggesting that the increased NO production did not result from inducible NOS expression. The present study demonstrates the successful in vivo transfer and functional expression of recombinant eNOS gene in large cerebral arteries. It also suggests that perivascular eNOS gene delivery via the CSF is a feasible approach that does not require interruption of cerebral blood flow. PMID- 9356492 TI - The affected gene underlying the class K glycosylphosphatidylinositol (GPI) surface protein defect codes for the GPI transamidase. AB - The final step in glycosylphosphatidylinositol (GPI) anchoring of cell surface proteins consists of a transamidation reaction in which preassembled GPI donors are substituted for C-terminal signal sequences in nascent polypeptides. In previous studies we described a human K562 cell mutant, termed class K, that accumulates fully assembled GPI units but is unable to transfer them to N terminally processed proproteins. In further work we showed that, unlike wild type microsomes, microsomes from these cells are unable to support C-terminal interaction of proproteins with the small nucleophiles hydrazine or hydroxylamine, and that the cells thus are defective in transamidation. In this study, using a modified recombinant vaccinia transient transfection system in conjunction with a composite cDNA prepared by 5' extension of an existing GenBank sequence, we found that the genetic element affected in these cells corresponds to the human homolog of yGPI8, a gene affected in a yeast mutant strain exhibiting similar accumulation of GPI donors without transfer. hGPI8 gives rise to mRNAs of 1.6 and 1.9 kb, both encoding a protein of 395 amino acids that varies in cells with their ability to couple GPIs to proteins. The gene spans approximately 25 kb of DNA on chromosome 1. Reconstitution of class K cells with hGPI8 abolishes their accumulation of GPI precursors and restores C-terminal processing of GPI-anchored proteins. Also, hGPI8 restores the ability of microsomes from the mutant cells to yield an active carbonyl in the presence of a proprotein which is considered to be an intermediate in catalysis by a transamidase. PMID- 9356491 TI - Reduction of HIV-1 in blood and lymph nodes following potent antiretroviral therapy and the virologic correlates of treatment failure. AB - Potent antiretroviral therapy can reduce plasma HIV RNA levels below the threshold of detection for periods of a year or more. The magnitude of HIV RNA reduction in the lymphoid tissue in patients with suppression of HIV RNA levels in plasma beyond 6 months has not been determined. We evaluated levels of HIV RNA and DNA and characterized resistance mutations in blood and inguinal lymph node biopsies obtained from 10 HIV-infected subjects who received 36-52 weeks of indinavir (IDV)/zidovudine (ZDV)/lamivudine (3TC), IDV, or ZDV/3TC. After 1 year of therapy, viral RNA levels in LN of individuals remained detectable but were log10 = 4 lower than in subjects on the triple drug regimen with interruption of therapy or in those treated with ZDV/3TC alone, who had viral loads in their lymph nodes indistinguishable from those expected for untreated patients. In all cases viral DNA remained detectable in lymph nodes and peripheral blood mononuclear cells (PBMC). When plasma virus suppression was incomplete, lymph node and PBMC cultures were positive and drug resistance developed. These studies indicate that pronounced and sustained suppression of plasma viremia by a potent antiretroviral combination is associated with low HIV RNA levels in the lymph nodes 1 year after treatment. Conversely, the persistence of even modest levels of plasma virus after 1 year of treatment reflects ongoing viral replication, the emergence of drug resistance, and the maintenance of high burdens of virus in the lymph nodes. PMID- 9356493 TI - Synergistic inhibition of human cancer cell growth by cytotoxic drugs and mixed backbone antisense oligonucleotide targeting protein kinase A. AB - Protein kinase A type I plays a key role in neoplastic transformation, conveying mitogenic signals of different growth factors and oncogenes. Inhibition of protein kinase A type I by antisense oligonucleotides targeting its RIalpha regulatory subunit results in cancer cell growth inhibition in vitro and in vivo. A novel mixed backbone oligonucleotide HYB 190 and its mismatched control HYB 239 were tested on soft agar growth of several human cancer cell types. HYB 190 demonstrated a dose-dependent inhibition of colony formation in all cell lines whereas the HYB 239 at the same doses caused a modest or no growth inhibition. A noninhibitory dose of each mixed backbone oligonucleotide was used in OVCAR-3 ovarian and GEO colon cancer cells to study whether any cooperative effect may occur between the antisense and a series of cytotoxic drugs acting by different mechanisms. Treatment with HYB 190 resulted in an additive growth inhibitory effect with several cytotoxic drugs when measured by soft agar colony formation. A synergistic growth inhibition, which correlated with increased apoptosis, was observed when HYB 190 was added to cancer cells treated with taxanes, platinum based compounds, and topoisomerase II selective drugs. This synergistic effect was also observed in breast cancer cells and was obtained with other related drugs such as docetaxel and carboplatin. Combination of HYB 190 and paclitaxel resulted in an accumulation of cells in late S-G2 phases of cell cycle and marked induction of apoptosis. A cooperative effect of HYB 190 and paclitaxel was also obtained in vivo in nude mice bearing human GEO colon cancer xenografts. These results are the first report of a cooperative growth inhibitory effect obtained in a variety of human cancer cell lines by antisense mixed backbone oligonucleotide targeting protein kinase A type I-mediated mitogenic signals and specific cytotoxic drugs. PMID- 9356494 TI - The Epstein-Barr virus oncogene product latent membrane protein 1 engages the tumor necrosis factor receptor-associated death domain protein to mediate B lymphocyte growth transformation and activate NF-kappaB. AB - The Epstein-Barr virus latent membrane protein 1 (LMP1) is essential for the transformation of B lymphocytes into lymphoblastoid cell lines. Previous data are consistent with a model that LMP1 is a constitutively activated receptor that transduces signals for transformation through its carboxyl-terminal cytoplasmic tail. One transformation effector site (TES1), located within the membrane proximal 45 residues of the cytoplasmic tail, constitutively engages tumor necrosis factor receptor-associated factors. Signals from TES1 are sufficient to drive initial proliferation of infected resting B lymphocytes, but most lymphoblastoid cells infected with a virus that does not express the 155 residues beyond TES1 fail to grow as long-term cell lines. We now find that mutating two tyrosines to an isoleucine at the carboxyl end of the cytoplasmic tail cripples the ability of EBV to cause lymphoblastoid cell outgrowth, thereby marking a second transformation effector site, TES2. A yeast two-hybrid screen identified TES2 interacting proteins, including the tumor necrosis factor receptor associated death domain protein (TRADD). TRADD was the only protein that interacted with wild-type TES2 and not with isoleucine-mutated TES2. TRADD associated with wild-type LMP1 but not with isoleucine-mutated LMP1 in mammalian cells, and TRADD constitutively associated with LMP1 in EBV-transformed cells. In transfection assays, TRADD and TES2 synergistically mediated high-level NF-kappaB activation. These results indicate that LMP1 appropriates TRADD to enable efficient long-term lymphoblastoid cell outgrowth. High-level NF-kappaB activation also appears to be a critical component of long-term outgrowth. PMID- 9356495 TI - Binding of factor VIIa to tissue factor induces alterations in gene expression in human fibroblast cells: up-regulation of poly(A) polymerase. AB - Tissue factor (TF) is the cellular receptor for an activated form of clotting factor VII (VIIa) and the binding of factor VII(a) to TF initiates the coagulation cascade. Sequence and structural patterns extracted from a global alignment of TF confers homology with interferon receptors of the cytokine receptor super family. Several recent studies suggested that TF could function as a genuine signal transducing receptor. However, it is unknown which biological function(s) of cells are altered upon the ligand, VIIa, binding to TF. In the present study, we examined the effect of VIIa binding to cell surface TF on cellular gene expression in fibroblasts. Differential mRNA display PCR technique was used to identify transcriptional changes in fibroblasts upon VIIa binding to TF. The display showed that VIIa binding to TF either up or down-regulated several mRNA species. The differential expression of one such transcript, VIIa induced up-regulation, was confirmed by Northern blot analysis. Isolation of a full-length cDNA corresponding to the differentially expressed transcript revealed that VIIa-up-regulated gene was poly(A) polymerase. Northern blot analysis of various carcinomas and normal human tissues revealed an over expression of PAP in cancer tissues. Enhanced expression of PAP upon VIIa binding to tumor cell TF may potentially play an important role in tumor metastasis. PMID- 9356496 TI - Transglutaminase-catalyzed inactivation of glyceraldehyde 3-phosphate dehydrogenase and alpha-ketoglutarate dehydrogenase complex by polyglutamine domains of pathological length. AB - Several adult-onset neurodegenerative diseases are caused by genes with expanded CAG triplet repeats within their coding regions and extended polyglutamine (Qn) domains within the expressed proteins. Generally, in clinically affected individuals n >/= 40. Glyceraldehyde 3-phosphate dehydrogenase binds tightly to four Qn disease proteins, but the significance of this interaction is unknown. We now report that purified glyceraldehyde 3-phosphate dehydrogenase is inactivated by tissue transglutaminase in the presence of glutathione S-transferase constructs containing a Qn domain of pathological length (n = 62 or 81). The dehydrogenase is less strongly inhibited by tissue transglutaminase in the presence of constructs containing shorter Qn domains (n = 0 or 10). Purified alpha-ketoglutarate dehydrogenase complex also is inactivated by tissue transglutaminase plus glutathione S-transferase constructs containing pathological-length Qn domains (n = 62 or 81). The results suggest that tissue transglutaminase-catalyzed covalent linkages involving the larger poly-Q domains may disrupt cerebral energy metabolism in CAG/Qn expansion diseases. PMID- 9356497 TI - A targeted mutation in the murine gene encoding the high density lipoprotein (HDL) receptor scavenger receptor class B type I reveals its key role in HDL metabolism. AB - Plasma high density lipoprotein (HDL), which protects against atherosclerosis, is thought to remove cholesterol from peripheral tissues and to deliver cholesteryl esters via a selective uptake pathway to the liver (reverse cholesterol transport) and steroidogenic tissues (e.g., adrenal gland for storage and hormone synthesis). Despite its physiologic and pathophysiologic importance, the cellular metabolism of HDL has not been well defined. The class B, type I scavenger receptor (SR-BI) has been proposed to play an important role in HDL metabolism because (i) it is a cell surface HDL receptor which mediates selective cholesterol uptake in cultured cells, (ii) its physiologically regulated expression is most abundant in the liver and steroidogenic tissues, and (iii) hepatic overexpression dramatically lowers plasma HDL. To test directly the normal role of SR-BI in HDL metabolism, we generated mice with a targeted null mutation in the SR-BI gene. In heterozygous and homozygous mutants relative to wild-type controls, plasma cholesterol concentrations were increased by approximately 31% and 125%, respectively, because of the formation of large, apolipoprotein A-I (apoA-I)-containing particles, and adrenal gland cholesterol content decreased by 42% and 72%, respectively. The plasma concentration of apoA I, the major protein in HDL, was unchanged in the mutants. This, in conjunction with the increased lipoprotein size, suggests that the increased plasma cholesterol in the mutants was due to decreased selective cholesterol uptake. These results provide strong support for the proposal that in mice the gene encoding SR-BI plays a key role in determining the levels of plasma lipoprotein cholesterol (primarily HDL) and the accumulation of cholesterol stores in the adrenal gland. If it has a similar role in controlling plasma HDL in humans, SR BI may influence the development and progression of atherosclerosis and may be an attractive candidate for therapeutic intervention in this disease. PMID- 9356498 TI - Inhibition of ubiquitin/proteasome-dependent protein degradation by the Gly-Ala repeat domain of the Epstein-Barr virus nuclear antigen 1. AB - The Epstein-Barr virus (EBV) encoded nuclear antigen (EBNA) 1 is expressed in latently infected B lymphocytes that persist for life in healthy virus carriers and is the only viral protein regularly detected in all EBV associated malignancies. The Gly-Ala repeat domain of EBNA1 was shown to inhibit in cis the presentation of major histocompatibility complex (MHC) class I restricted cytotoxic T cell epitopes from EBNA4. It appears that the majority of antigens presented via the MHC I pathway are subject to ATP-dependent ubiquitination and degradation by the proteasome. We have investigated the influence of the repeat on this process by comparing the degradation of EBNA1, EBNA4, and Gly-Ala containing EBNA4 chimeras in a cell-free system. EBNA4 was efficiently degraded in an ATP/ubiquitin/proteasome-dependent fashion whereas EBNA1 was resistant to degradation. Processing of EBNA1 was restored by deletion of the Gly-Ala domain whereas insertion of Gly-Ala repeats of various lengths and in different positions prevented the degradation of EBNA4 without appreciable effect on ubiquitination. Inhibition was also achieved by insertion of a Pro-Ala coding sequence. The results suggest that the repeat may affect MHC I restricted responses by inhibiting antigen processing via the ubiquitin/proteasome pathway. The presence of regularly interspersed Ala residues appears to be important for the effect. PMID- 9356499 TI - Cytokine-mediated enhancement of epidermal growth factor receptor expression provides an immunological approach to the therapy of pancreatic cancer. AB - The increased expression of epidermal growth factor receptor induced by tumor necrosis factor alpha renders pancreatic cancer cells more susceptible to antibody-dependent cellular cytotoxicity by a mAb specific for this receptor. Laboratory studies with athymic mice bearing xenografts of human pancreatic cancer cells demonstrated a cytokine-induced ability of the mAb to cause significant tumor regression. In a phase I/II clinical trial, 26 patients with unresectable pancreatic cancer were enrolled into three cohorts receiving variable amounts of the antibody together with a constant amount of tumor necrosis factor alpha. With increasing doses of antibody, the growth of the primary tumor was significantly inhibited. This was reflected by a longer median survival, with one complete remission lasting for 3 years obtained with the highest dose of antibody employed. Thus, a combination of the cytokine, tumor necrosis factor alpha, with a mAb to the epidermal growth factor receptor offers a potentially useful approach for the treatment of pancreatic cancer. PMID- 9356500 TI - Enhancement of DNA repair in human skin cells by thymidine dinucleotides: evidence for a p53-mediated mammalian SOS response. AB - Thymidine dinucleotide (pTpT) stimulates melanogenesis in mammalian pigment cells and intact skin, mimicking the effects of UV irradiation and UV-mimetic DNA damage. Here it is shown that, in addition to tanning, pTpT induces a second photoprotective response, enhanced repair of UV-induced DNA damage. This enhanced repair results in a 2-fold increase in expression of a UV-damaged chloramphenicol acetyltransferase expression vector transfected into pTpT-treated skin fibroblasts and keratinocytes, compared with diluent-treated cells. Direct measurement of thymine dimers and (6-4) photoproducts by immunoassay demonstrates faster repair of both of these UV-induced photoproducts in pTpT-treated fibroblasts. This enhanced repair capacity also improves cell survival and colony forming ability after irradiation. These effects of pTpT are accomplished, at least in part, by the up-regulation of a set of genes involved in DNA repair (ERCC3 and GADD45) and cell cycle inhibition (SDI1). At least two of these genes (GADD45 and SDI1) are known to be transcriptionally regulated by the p53 tumor suppressor protein. Here we show that pTpT activates p53, leading to nuclear accumulation of this protein, and also increases the specific binding of this transcription factor to its DNA consensus sequence. PMID- 9356501 TI - Archaeal nucleosomes. AB - Archaea contain histones that have primary sequences in common with eukaryal nucleosome core histones and a three-dimensional structure that is essentially only the histone fold. Here we report the results of experiments that document that archaeal histones compact DNA in vivo into structures similar to the structure formed by the histone (H3+H4)2 tetramer at the center of the eukaryal nucleosome. After formaldehyde cross-linking in vivo, these archaeal nucleosomes have been isolated from Methanobacterium thermoautotrophicum and Methanothermus fervidus, visualized by electron microscopy on plasmid and genomic DNAs, and shown by immunogold labeling, SDS/PAGE, and immunoblotting to contain archaeal histones, cross-linked into tetramers. Archaeal nucleosomes protect approximately 60 bp of DNA and multiples of approximately 60 bp from micrococcal nuclease digestion, and immunoprecipitation has demonstrated that most, but not all, M. fervidus genomic DNA sequences are associated in vivo with archaeal histones. PMID- 9356503 TI - Cerebral cortical astroglia from the trisomy 16 mouse, a model for down syndrome, produce neuronal cholinergic deficits in cell culture. AB - Trisomy 21 (Down syndrome) is associated with a high incidence of Alzheimer disease and with deficits in cholinergic function in humans. We used the trisomy 16 (Ts16) mouse model for Down syndrome to identify the cellular basis for the cholinergic dysfunction. Cholinergic neurons and cerebral cortical astroglia, obtained separately from Ts16 mouse fetuses and their euploid littermates, were cultured in various combinations. Choline acetyltransferase activity and cholinergic neuron number were both depressed in cultures in which both neurons and glia were derived from Ts16 fetuses. Cholinergic function of normal neurons was significantly down-regulated by coculture with Ts16 glia. Conversely, neurons from Ts16 animals could express normal cholinergic function when grown with normal glia. These observations indicate that astroglia may contribute strongly to the abnormal cholinergic function in the mouse Ts16 model for Down syndrome. The Ts16 glia could lack a cholinergic supporting factor present in normal glia or contain a factor that down-regulates cholinergic function. PMID- 9356502 TI - Yersinia enterocolitica induces apoptosis in macrophages by a process requiring functional type III secretion and translocation mechanisms and involving YopP, presumably acting as an effector protein. AB - Yersiniae, causative agents of plague and gastrointestinal diseases, secrete and translocate Yop effector proteins into the cytosol of macrophages, leading to disruption of host defense mechanisms. It is shown in this report that Yersinia enterocolitica induces apoptosis in macrophages and that this effect depends on YopP. Functional secretion and translocation mechanisms are required for YopP to act, strongly suggesting that this protein exerts its effect intracellularly, after translocation into the macrophages. YopP shows a high level of sequence similarity with AvrRxv, an avirulence protein from Xanthomonas campestris, a plant pathogen that induces programmed cell death in plant cells. This indicates possible similarities between the strategies used by pathogenic bacteria to elicit programmed cell death in both plant and animal hosts. PMID- 9356504 TI - Deciphering a neural code for vision. AB - Deciphering the information that eyes, ears, and other sensory organs transmit to the brain is important for understanding the neural basis of behavior. Recordings from single sensory nerve cells have yielded useful insights, but single neurons generally do not mediate behavior; networks of neurons do. Monitoring the activity of all cells in a neural network of a behaving animal, however, is not yet possible. Taking an alternative approach, we used a realistic cell-based model to compute the ensemble of neural activity generated by one sensory organ, the lateral eye of the horseshoe crab, Limulus polyphemus. We studied how the neural network of this eye encodes natural scenes by presenting to the model movies recorded with a video camera mounted above the eye of an animal that was exploring its underwater habitat. Model predictions were confirmed by simultaneously recording responses from single optic nerve fibers of the same animal. We report here that the eye transmits to the brain robust "neural images" of objects having the size, contrast, and motion of potential mates. The neural code for such objects is not found in ambiguous messages of individual optic nerve fibers but rather in patterns of coherent activity that extend over small ensembles of nerve fibers and are bound together by stimulus motion. Integrative properties of neurons in the first synaptic layer of the brain appear well suited to detecting the patterns of coherent activity. Neural coding by this relatively simple eye helps explain how horseshoe crabs find mates and may lead to a better understanding of how more complex sensory organs process information. PMID- 9356505 TI - Dynamic regulation of c-Jun N-terminal kinase activity in mouse brain by environmental stimuli. AB - Activation of the recently identified c-Jun N-terminal kinases (JNKs) typically results in programmed cell death (apoptosis) in neurons and other cell types grown in culture. However, the effects of JNK activation in the central nervous system in vivo are unknown. At baseline, JNK activity in mice was on average 17 fold higher in brain than in peripheral organs, whereas JNK protein levels were similar. In brain, JNK was expressed primarily in neurons. Restraining mice or allowing them to explore a novel environment rapidly increased JNK activity 3- to 15-fold in various brain regions, but these manipulations did not increase brain activity of the extracellular signal-regulated kinase. Because noninvasive environmental stimuli that do not induce neurodegeneration elicited prominent increases in JNK activity in the brain, we conclude that acute activation of the JNK cascade in central nervous system neurons does not induce neuronal apoptosis in vivo. In contrast, the high baseline activity of JNK in the brain and the activation of the JNK cascade by environmental stimuli suggest that this kinase may play an important physiological role in neuronal function. PMID- 9356506 TI - Localization of neuropeptide Y Y1 receptors in cerebral blood vessels. AB - The localization of neuropeptide Y (NPY) Y1 receptor (R) -like immunoreactivity (LI) has been studied in cerebral arteries and arterioles of the rat by immunohistochemistry using fluorescence, confocal, and electron microscopy. High levels of Y1-R-LI were observed in smooth muscle cells (SMCs) in the small arterioles of the pial arterial network, especially on the basal surface of the brain, and low levels in the major basal cerebral arteries. The levels of Y1-R-LI varied strongly between adjacent SMCs. Y1-R-LI was associated with small endocytosis vesicles, mainly on the outer surface of the SMCs, but also on their endothelial side and often laterally at the interface between two SMCs. NPY immunoreactive (Ir) nerve fibers could not be detected in association with the Y1 R-rich small arterioles but only around arteries with low Y1-R levels. A dense network of central NPY-Ir nerve fibers in the superficial layers of the brain was lying close to the strongly Y1-R-Ir small arterioles. The results indicate that NPY has a profound effect on small arterioles of the brain acting on Y1-Rs, both on the peripheral and luminal side of the SMCs. However, the source of the endogenous ligand, NPY, remains unclear. NPY released from central neurons may play a role, in addition to blood-borne NPY. PMID- 9356507 TI - Effects of muscarinic blockade in perirhinal cortex during visual recognition. AB - Stimulus recognition in monkeys is severely impaired by destruction or dysfunction of the perirhinal cortex and also by systemic administration of the cholinergic-muscarinic receptor blocker, scopolamine. These two effects are shown here to be linked: Stimulus recognition was found to be significantly impaired after bilateral microinjection of scopolamine directly into the perirhinal cortex, but not after equivalent injections into the laterally adjacent visual area TE or into the dentate gyrus of the overlying hippocampal formation. The results suggest that the formation of stimulus memories depends critically on cholinergic-muscarinic activation of the perirhinal area, providing a new clue to how stimulus representations are stored. PMID- 9356508 TI - Identification of genes differentially expressed by nerve growth factor- and neurotrophin-3-dependent sensory neurons. AB - The neurotrophins nerve growth factor (NGF) and neurotrophin-3 (NT3) support the survival of subpopulations of primary sensory neurons with defined and distinct physiological characteristics. Only a few genes have been identified as being differentially expressed in these subpopulations, and not much is known about the nature of the molecules involved in the processing of sensory information in NGF dependent nociceptive neurons or NT3-dependent proprioceptive neurons. We devised a simple dorsal root ganglion (DRG) explant culture system, allowing the selection of neuronal populations preferentially responsive to NGF or NT3. The reliability of this assay was first monitored by the differential expression of the NGF and NT3 receptors trkA and trkC, as well as that of neuropeptides and calcium-binding proteins. We then identified four differentially expressed sodium channels, two enriched in the NGF population and two others in the NT3 population. Finally, using an optimized RNA fingerprinting protocol, we identified 20 additional genes, all differentially expressed in DRG explants cultured with NGF or NT3. This approach thus allows the identification of large number of genes expressed in subpopulations of primary sensory neurons and opens the possibility of studying the molecular mechanisms of nociception and proprioception. PMID- 9356509 TI - Ultrastructural localization of zinc transporter-3 (ZnT-3) to synaptic vesicle membranes within mossy fiber boutons in the hippocampus of mouse and monkey. AB - Zinc transporter-3 (ZnT-3), a member of a growing family of mammalian zinc transporters, is expressed in regions of the brain that are rich in histochemically reactive zinc (as revealed by the Timm's stain), including entorhinal cortex, amygdala, and hippocampus. ZnT-3 protein is most abundant in the zinc-enriched mossy fibers that project from the dentate granule cells to hilar and CA3 pyramidal neurons. We show here by electron microscopy that ZnT-3 decorates the membranes of all clear, small, round synaptic vesicles (SVs) in the mossy fiber boutons of both mouse and monkey. Furthermore, up to 60-80% of these SVs contain Timm's-stainable zinc. The coincidence of ZnT-3 on the membranes of SVs that accumulate zinc, and its homology with known zinc transporters, suggest that ZnT-3 is responsible for the transport of zinc into SVs, and hence for the ability of these neurons to release zinc upon excitation. PMID- 9356510 TI - Association of INAD with NORPA is essential for controlled activation and deactivation of Drosophila phototransduction in vivo. AB - Visual transduction in Drosophila is a G protein-coupled phospholipase C-mediated process that leads to depolarization via activation of the transient receptor potential (TRP) calcium channel. Inactivation-no-afterpotential D (INAD) is an adaptor protein containing PDZ domains known to interact with TRP. Immunoprecipitation studies indicate that INAD also binds to eye-specific protein kinase C and the phospholipase C, no-receptor-potential A (NORPA). By overlay assay and site-directed mutagenesis we have defined the essential elements of the NORPA-INAD association and identified three critical residues in the C-terminal tail of NORPA that are required for the interaction. These residues, Phe-Cys-Ala, constitute a novel binding motif distinct from the sequences recognized by the PDZ domain in INAD. To evaluate the functional significance of the INAD-NORPA association in vivo, we generated transgenic flies expressing a modified NORPA, NORPAC1094S, that lacks the INAD interaction. The transgenic animals display a unique electroretinogram phenotype characterized by slow activation and prolonged deactivation. Double mutant analysis suggests a possible inaccessibility of eye specific protein kinase C to NORPAC1094S, undermining the observed defective deactivation, and that delayed activation may similarly result from NORPAC1094S being unable to localize in close proximity to the TRP channel. We conclude that INAD acts as a scaffold protein that facilitates NORPA-TRP interactions required for gating of the TRP channel in photoreceptor cells. PMID- 9356511 TI - Degeneration of neurons, synapses, and neuropil and glial activation in a murine Atm knockout model of ataxia-telangiectasia. AB - Neural degeneration is one of the clinical manifestations of ataxia telangiectasia, a disorder caused by mutations in the Atm protein kinase gene. However, neural degeneration was not detected with general purpose light microscopic methods in previous studies using several different lines of mice with disrupted Atm genes. Here, we show electron microscopic evidence of degeneration of several different types of neurons in the cerebellar cortex of 2 month-old Atm knockout mice, which is accompanied by glial activation, deterioration of neuropil structure, and both pre- and postsynaptic degeneration. These findings are similar to those in patients with ataxia-telangiectasia, indicating that Atm knockout mice are a useful model to elucidate the mechanisms underlying neurodegeneration in this condition and to develop and test strategies to palliate and prevent the disease. PMID- 9356512 TI - Acoustic and neural bases for innate recognition of song. AB - In behavior reminiscent of the responsiveness of human infants to speech, young songbirds innately recognize and prefer to learn the songs of their own species. The acoustic and physiological bases for innate recognition were investigated in fledgling white-crowned sparrows lacking song experience. A behavioral test revealed that the complete conspecific song was not essential for innate recognition: songs composed of single white-crowned sparrow phrases and songs played in reverse elicited vocal responses as strongly as did normal song. In all cases, these responses surpassed those to other species' songs. Although auditory neurons in the song nucleus HVc and the underlying neostriatum of fledglings did not prefer conspecific song over foreign song, some neurons responded strongly to particular phrase types characteristic of white-crowned sparrows and, thus, could contribute to innate song recognition. PMID- 9356513 TI - Synchronization of oscillatory responses in visual cortex correlates with perception in interocular rivalry. AB - In subjects suffering from early onset strabismus, signals conveyed by the two eyes are not perceived simultaneously but in alternation. We exploited this phenomenon of interocular suppression to investigate the neuronal correlate of binocular rivalry in primary visual cortex of awake strabismic cats. Monocularly presented stimuli that were readily perceived by the animal evoked synchronized discharges with an oscillatory patterning in the gamma-frequency range. Upon dichoptic stimulation, neurons responding to the stimulus that continued to be perceived increased the synchronicity and the regularity of their oscillatory patterning while the reverse was true for neurons responding to the stimulus that was no longer perceived. These differential changes were not associated with modifications of discharge rate, suggesting that at early stages of visual processing the degree of synchronicity rather than the amplitude of responses determines which signals are perceived and control behavioral responses. PMID- 9356514 TI - Direct measurement of nitric oxide generation from nitric oxide synthase. AB - Although nitric oxide synthase (NOS) is widely considered as the major source of NO in biological cells and tissues, direct evidence demonstrating NO formation from the purified enzyme has been lacking. It was recently reported that NOS does not synthesize NO, but rather generates nitroxyl anion (NO-) that is subsequently converted to NO by superoxide dismutase (SOD). To determine if NOS synthesizes NO, electron paramagnetic resonance (EPR) spectroscopy was applied to directly measure NO formation from purified neuronal NOS. In the presence of the NO trap Fe2+-N-methyl-D-glucamine dithiocarbamate, NO gives rise to characteristic EPR signals with g = 2.04 and aN = 12.7 G, whereas NO- is undetectable. In the presence of L-arginine (L-Arg) and cofactors, NOS generated prominent NO signals. This NO generation did not require SOD, and it was blocked by the specific NOS inhibitor N-nitro-L-arginine methyl ester. Isotope-labeling experiments with L [15N]Arg further demonstrated that NOS-catalyzed NO arose from the guanidino nitrogen of L-Arg. Measurement of the time course of NO formation demonstrated that it paralleled that of L-citrulline. The conditions used in the prior study were shown to result in potent superoxide generation, and this may explain the failure to measure NO formation in the absence of SOD. These experiments provide unequivocal evidence that NOS does directly synthesize NO from L-Arg. PMID- 9356518 TI - Ethnohistory, genetics, and cancer mortality in Europeans. AB - Geographic variation in cancer rates is thought to be the result of two major factors: environmental agents varying spatially and the attributes, genetic or cultural, of the populations inhabiting the areas studied. These attributes in turn result from the history of the populations in question. We had previously constructed an ethnohistorical database for Europe since 2200 B.C., permitting estimates of the ethnic composition of modern European populations. We were able to show that these estimates correlate with genetic distances. In this study, we wanted to see whether they also correlate with cancer rates. We employed two data sets of cancer mortalities from 42 types of cancer for the European Economic Community and for Central Europe. We subjected spatial differences in cancer mortalities, genetic, ethnohistorical, and geographic distances to matrix permutation tests to determine the magnitude and significance of their association. Our findings are that distances in cancer mortalities are correlated more with ethnohistorical distances than with genetic distances. Possibly the cancer rates may be affected by loci other than the genetic systems available to us, and/or by cultural factors mediated by the ethnohistorical differences. We find it remarkable that patterns of frequently ancient ethnic admixture are still reflected in modern cancer mortalities. Partial correlations with geography suggest that local environmental factors affect the mortalities as well. PMID- 9356517 TI - Efficient gene tagging in Arabidopsis thaliana using a gene trap approach. AB - Large quantities of DNA sequence information about plant genes are rapidly accumulating in public databases, but to progress from DNA sequence to biological function a mutant allele for each of the genes ideally should be available. Here we describe a gene trap construct that allowed us to disrupt transcribed genes with a high efficiency in Arabidopsis thaliana. In the T-DNA vector used, the expression of a bacterial reporter gene coding for neomycin phosphotransferase II (nptII) depends on the in vivo generation of a translation fusion upon the T-DNA integration into the Arabidopsis genome. Analysis of 20 selected transgenic lines showed that 12 lines are T-DNA insertion mutants. The disrupted genes analyzed encoded ribosomal proteins (three lines), aspartate tRNA synthase, DNA ligase, basic-domain leucine zipper DNA binding protein, ATP-binding cassette transporter, and five proteins of unknown function. Four tagged genes were new for Arabidopsis. The results presented here suggest that gene trapping, using nptII as a reporter gene, can be as high as 80% and opens novel perspectives for systematic gene tagging in A. thaliana. PMID- 9356519 TI - Vitamin C pharmacokinetics: it's deja vu all over again. PMID- 9356520 TI - Distinguishing malnutrition from disease: the search goes on. PMID- 9356521 TI - Dairy products and colon cancer: mechanisms of the protective effect. PMID- 9356522 TI - Functional foods in the prevention and treatment of disease: significance of the Dietary Approaches to Stop Hypertension Study. PMID- 9356523 TI - Dietary carbohydrates and insulin sensitivity: a review of the evidence and clinical implications. AB - Insulin resistance is associated with diabetes mellitus, ischemic heart disease, and hypertension both independently and as part of syndrome X. Environmental influences on SI are incompletely understood. Exercise has a strong beneficial effect and obesity a strong adverse effect. The balance of evidence suggests that a high-fat diet is likely to reduce insulin sensitivity but the effects of dietary carbohydrates are more controversial. Extensive studies in animals showed a detrimental effect of diets very high in fructose or sucrose, particularly in association with induction of hypertriglyceridemia. The more limited studies in humans had conflicting results, partly because of heterogeneity of design. Certain groups of subjects may be more sensitive to adverse effects of high intakes of dietary sucrose or fructose. More carefully controlled studies in humans are needed to provide evidence on which to base public health policies with respect to dietary carbohydrates and SI. PMID- 9356525 TI - Body mass index and height from childhood to adulthood in the 1958 British born cohort. AB - The purpose of this study was to assess relations among height, weight, and body mass index (BMI) at different ages from childhood to adulthood, and to examine long-term relations among timing of puberty, height, and BMI. Longitudinal data from the 1958 British birth cohort (all children born between March 3rd and 9th, 1958) were used. Height and weight were measured at ages 7, 11, 16, 23 (self reported), and 33 y; pubertal status was assessed at ages 11 and 16 y. Data for 5700 females and 5512 males were analyzed. Adult height was well predicted from childhood, with strong correlations (r = 0.7 for both sexes) between height at ages 7 and 33 y. Correlations for BMI were weaker, especially between childhood and early adulthood (r = 0.33 for males and 0.37 for females, ages 7 and 33 y), although they increased with increasing age. Although the fattest children had the highest risks of adult obesity, most obese adults had not been fat at earlier ages: only 17% and 18% of obese 33-y-old men and women, respectively, had been fat at age 7 y. A strong and evenly graded association was found between timing of puberty and BMI, with higher mean BMIs for the earlier maturers at ages 7-33 y. The moderate prediction of adult BMI in this large and unselected sample suggests that although the prevention of childhood fatness may be desirable, most obese adults could not be identified from their childhood BMI, and hence, preventive strategies need to be population-based. PMID- 9356524 TI - Basal metabolism of weight-stable chronically undernourished men and women: lack of metabolic adaptation and ethnic differences. AB - The purpose of this study was to investigate whether weight-stable chronically energy-deficient subjects exhibit evidence of metabolic adaptation and to establish whether international predictive equations overestimate the basal metabolic rate (BMR) of tropical populations. BMR, body weight, height, and fat free mass (FFM) by underwater weighing were measured in healthy, physically active urban dwellers of low socioeconomic status (178 men and women aged 22-38 y) in Bangalore, Southern India. Subjects were selected on the basis of body mass index (BMI; in kg/m2) and classified in three groups: severely undernourished (BMI < 17.0; n = 30 men, n = 25 women), marginally undernourished (BMI = 17.0 18.5; n = 31 men, n = 30 women), and well nourished (BMI > 18.5; n = 27 men, n = 35 women). The BMR of the well-nourished group, expressed in absolute terms (6.20 and 5.18 MJ/d for men and women, respectively), was significantly higher (P < 0.000) than that of the severely undernourished group (5.72 and 4.64 MJ/d for men and women, respectively). Normalizing BMR for either body weight or FFM by analysis of covariance abolished all differences. The mean BMR of the low-BMI study group was substantially higher (11-14%) than reported previously for undernourished Indian adults. The BMR of both men and women, regardless of their nutritional status, was accurately estimated by age- and sex-specific FAO/WHO/UNU equations. These findings suggest the absence of an enhanced metabolic response in weight-stable chronically undernourished adults. This is in contrast with earlier reports, and supports more recent views. The study also provides evidence of the absence of ethnic-specific energy turnover in Indians. PMID- 9356527 TI - Thermic effect of food during each phase of the menstrual cycle. AB - The effect of the menstrual cycle on the thermic effect of food (TEF) was examined in eight healthy, normal-weight [mean +/- SD: 56.1 +/- 5.6 kg and body mass index (in kg/m2) 21.3 +/- 1.8] women aged 22-38 y. Their lean body mass and fat mass were 39.4 +/- 2.7 kg and 16.9 +/- 6.5 kg, respectively. TEF was measured on 4 separate days selected to match the four phases of a menstrual cycle: early follicular, follicular, luteal, and late luteal. The volunteers consumed a 3138 kJ liquid meal (54.5% carbohydrate, 14.0% protein, and 31.5% fat) on each test day. Resting metabolic rate was measured for 55 min before the meal and every 30 min after the start of the meal for 205 min. Although resting metabolic rate remained unchanged, there was a significant difference (P < 0.01 by ANOVA) in mean (+/- SEM) values for TEF among the four phases of the cycle: 0.94 +/- 0.05 kJ/min during the early follicular phase, 0.86 +/- 0.09 kJ/min during the follicular phase, 0.70 +/- 0.10 kJ/min during the luteal phase, and 0.76 +/- 0.07 kJ/min during the late luteal phase. TEF decreased significantly (P < 0.025 by paired t test) during postovulation (average of luteal and late luteal phases), when it was 0.73 +/- 0.07 kJ/min, compared with preovulation (average of early follicular and follicular phases), when it was 0.90 +/- 0.06 kJ/min. In conclusion, TEF decreased during the luteal phase of the menstrual cycle, possibly as a result of impairment of glucose uptake and slower transit of food through the upper gastrointestinal tract. PMID- 9356526 TI - Breast milk or animal-product foods improve linear growth of Peruvian toddlers consuming marginal diets. AB - Although breast-feeding is widely accepted as important for infant health, its benefits during the second year of life have been questioned. We analyzed data from 107 breast-fed and weaned Peruvian children living in a periurban community to determine whether breast milk contributed to improved linear growth between 12 and 15 mo of age. Breast-feeding frequency was self-reported; intakes of complementary foods and animal products were estimated from a food-frequency survey. Multivariate-linear-regression analysis was used to predict the length of the children at 15 mo of age. Determinants of length included length and weight for-length at 12 mo of age (US National Center for Health Statistics standards), interval between 12- and 15-mo measurements, breast-feeding frequency, incidence of diarrhea, and intakes of complementary and animal-product foods. Complementary foods, animal-product foods, and breast milk all promoted toddlers' linear growth. In subjects with low intakes of animal-product foods, breast-feeding was positively associated (P < 0.05) with linear growth. There was a 0.5-cm/3 mo difference in linear growth between weaned toddlers and children who consumed the average number of feedings of breast milk. Linear growth was also positively associated with intake of animal-product foods in children with low intakes of complementary foods. The negative association between diarrhea and linear growth did not occur in subjects with high complementary-food intakes. When the family's diet is low in quality, breast milk is an especially important source of energy, protein, and accompanying micronutrients in young children. Thus, continued breast-feeding after 1 y of age, in conjunction with feeding of complementary foods, should be encouraged in toddlers living in poor circumstances. PMID- 9356528 TI - Lactation and weight retention. AB - The effect of lactation on weight retention was investigated longitudinally, with data collected at 0.5, 2, 4, 6, 12, and 18 mo after parturition in 110 women aged 20-40 y who had been nulliparous or primiparous. At each evaluation women were categorized as fully breast-feeding, partly breast-feeding, or bottle-feeding including infants weaned to a bottle (bottle feeding/weaned). Postpartum weight retention was calculated by subtracting weight before pregnancy from weight at each evaluation. Lactation practices were found to be significantly associated (P < 0.05) with postpartum weight retention by longitudinal regression analysis. Women who bottle-fed their infants retained more weight over time than women who breast-fed their infants. Significantly slower rates of weight loss were observed when women ceased breast-feeding or switched from fully to partly breast-feeding. Weight retention over time was greater in women who were older, unmarried, or had greater weight gain during pregnancy (P < 0.05). A pattern of weight gain rather than weight loss was observed in unmarried women. Our findings suggest that lactation influences the pattern of postpartum weight retention; however, the effect of lactation on weight retention was sufficiently limited to warrant minimal emphasis on lactation as a means of minimizing postpartum weight retention. PMID- 9356529 TI - Consumption of green or black tea does not increase resistance of low-density lipoprotein to oxidation in humans. AB - Epidemiologic studies indicated that tea consumption reduces the risk of cardiovascular disease. We assessed the effect of green or black tea consumption on resistance of low-density lipoprotein (LDL) to oxidation ex vivo and on serum lipid concentrations in healthy volunteers. In a 4-wk parallel comparison trial, 45 volunteers consumed 900 mL (6 cups) mineral water, green tea, or black tea/d. Blood samples drawn while subjects were fasting were obtained before and after the study. The effect on resistance of subsequently isolated LDL to oxidation of adding green or black tea extract to plasma was investigated in an in vitro experiment. Consumption of 900 mL (6 cups) green or black tea/d did not affect serum lipid concentrations, resistance of LDL to oxidation, or markers of oxidative damage to lipids in vivo, although consumption of green tea slightly increased total antioxidant activity of plasma. The in vitro experiment showed that resistance of isolated LDL to oxidation increased only after incubation of plasma with very high amounts of green or black tea. These amounts, when converted to tea catechin concentrations, were much higher than those expected in vivo. We conclude that daily consumption of 900 mL (6 cups) green or black tea/d for 4 wk had no effect on serum lipid concentrations or resistance of LDL to oxidation ex vivo. Future research should focus on mechanisms by which tea flavonoids may reduce the risk of cardiovascular disease other than by increasing the intrinsic antioxidant status of LDL. PMID- 9356530 TI - Interactions in the postprandial appearance of beta-carotene and canthaxanthin in plasma triacylglycerol-rich lipoproteins in humans. AB - We investigated the plasma appearance of beta-carotene and canthaxanthin, an oxycarotenoid, in normolipidemic premenopausal women (n = 9) who ingested beta carotene alone, canthaxanthin alone, and a combined dose. Blood samples were taken hourly for 12 h; additional blood samples were collected over 528 h. In a subset of the women (n = 5), plasma lipoproteins were separated into chylomicrons, very-low-density-lipoproteins (VLDL) subfractions, intermediate density lipoproteins (IDLs), and low-density lipoproteins (LDLs). The appearance of beta-carotene in plasma was biphasic, with a minor peak at 5 h followed by a sustained peak at 24-48 h. The plasma appearance of canthaxanthin was monophasic, with a rapid increase to the final hourly measurement at 12 h and a steady decrease from the next measurement at 24 h. At 6 h, 23.4 +/- 2.9% of the increase in plasma canthaxanthin was associated with LDL, in contrast with 2.4 +/- 1.4% of the increase in plasma beta-carotene (P < 0.005). Ingestion of a combined dose of beta-carotene and canthaxanthin inhibited the appearance of canthaxanthin in plasma, chylomicrons, and each VLDL subfraction (P < 0.05), but did not significantly affect the rapid accumulation of canthaxanthin in LDL within 10 h. In contrast, ingestion of the combined dose did not significantly affect the appearance of beta-carotene in plasma or plasma lipoproteins. These findings suggest distinct mechanisms of incorporation into lipoproteins and specific interactions of beta-carotene and canthaxanthin during intestinal absorption in humans. PMID- 9356533 TI - Effect of hemolysis on serum retinol as assessed by direct fluorometry. AB - To assess the effect of hemolysis on serum retinol concentrations determined by direct fluorometry, we assayed 196 blood samples from children 6-72-mo of age with various grades of hemolysis for serum retinol by both fluorescence and HPLC. Mean serum retinol concentrations determined by HPLC did not differ significantly according to hemolysis grade; however, fluorometric values did. Additionally, serum retinol concentrations obtained from HPLC and those obtained from direct fluorometry were significantly different in samples with severe hemolysis. Multivariate-regression analysis showed that hemolysis grade was a significant predictor of the difference in mean serum retinol values determined by the two methods. Although severe hemolysis interfered with determinations of serum retinol by direct fluorometry, this method is still a viable choice for field studies of vitamin A status. PMID- 9356531 TI - Effect of an acute reduction in carbohydrate intake on subsequent food intake in healthy men. AB - We investigated the effect of a combined carbohydrate and energy deficit in the regulation of food intake during 1 d. Seven lean, male subjects were studied in a crossover design. After 7 d of consuming a baseline diet (40% of energy as fat, 45% as carbohydrate, and 15% as protein), subjects were deprived of carbohydrate for 24 h; baseline amounts of fat and protein were consumed but only one-third of the baseline amount of carbohydrate. On the following outcome day, subjects were free to select ad libitum from a selection of either high-carbohydrate or low carbohydrate food. On the baseline diet subjects consumed on average 10.9 +/- 1.7 MJ/d (carbohydrate: 305 +/- 49 g/d; fat: 116 +/- 18 g/d) and there was no difference in baseline intake between the two phases of the crossover study. During the deficit day, intake was reduced to 7.7 +/- 1.2 MJ/d [carbohydrate: 110 +/- 25 g/d (66% reduction); fat: 116 +/- 18 g/d]. On the outcome day, energy intake from high-carbohydrate foods was on average 10.5 MJ/d (carbohydrate: 430 +/- 112 g/d; fat: 48 +/- 20 g/d) compared with 16.6 MJ/d from high-fat foods (carbohydrate: 312 +/- 84 g/d; fat: 258 +/- 78 g/d). We conclude that the restoration of an energy deficit is not the main factor determining acute food intake. Rather, the data support the hypothesis that, under the conditions of our experiment, the intake of carbohydrate required to maintain carbohydrate balance was a more important factor in the regulation of acute food intake than was the restoration of energy deficit is not. PMID- 9356534 TI - Pharmacokinetic perspectives on megadoses of ascorbic acid. AB - Ascorbic acid (vitamin C) is commonly used as a dietary supplement, often in megadoses. However, as the daily oral dose is increased, the concentration of ascorbic acid in the plasma and other body fluids does not increase proportionally, but instead tends to approach an upper limit. For example, when the daily dose is increased from 200 to 2500 mg (from 1.1 to 14.2 mmol) the mean steady state plasma concentration increases only from approximately 12 to 15 mg/L (from 68.1 to 85.2 mumol/L). Published data were reanalyzed with an integrated modeling approach to shed new quantitative light on this phenomenon. This analysis is based on the renal clearance of ascorbic acid, which rises sharply with increasing plasma concentrations as a result of saturable tubular reabsorption. The analysis indicates that both saturable gastrointestinal absorption and nonlinear renal clearance act additively to produce the ceiling effect in plasma concentrations. As a consequence of this ceiling effect, there is no pharmacokinetic justification for the use of megadoses of ascorbic acid. PMID- 9356532 TI - Metabolic effects of digestible and partially indigestible cornstarch: a study in the absorptive and postabsorptive periods in healthy humans. AB - To compare the effects of digestible (pregelatinized) and partially indigestible (retrograded) cornstarches on some metabolic indexes, we studied eight healthy volunteers during two periods separated by 1 wk. In each period, fasting volunteers consumed at 0800 the test meal containing either the digestible or partially indigestible cornstarch; blood and breath were sampled in the absorptive period for 8 h. To study its late effects, the same test meal as that served at 0800 was given again at 2200, and blood and breath were sampled for 3 h in the postabsorptive period the next morning, i.e., 10 h after ingestion of the test meal. In the absorptive period, blood glucose and insulin were significantly higher after ingestion of digestible cornstarch than after partially indigestible cornstarch. In the postabsorptive period concentrations of blood glucose, insulin, and fatty acids were not significantly different, whereas concentrations of blood acetate, breath hydrogen, methane, and 13CO2, and the respiratory quotient and satiety were significantly higher (P < 0.05) and concentrations of blood glycerol significantly lower (P < 0.05) after ingestion of partially indigestible cornstarch than after digestible cornstarch. We conclude that in healthy humans, digestion of partially indigestible cornstarch is slow in the small intestine and its colonic fermentation continues 10-13 h after its ingestion. Compared with pregelatinized cornstarch, the shift in starch digestion induced by retrogradation leads to a reduction in glycemic and insulinemic responses in the absorptive period and in lipolysis in the postabsorptive. PMID- 9356535 TI - Calcium and magnesium balance in 9-14-y-old children. AB - Few data are available regarding calcium and magnesium absorption and endogenous fecal excretion in children. We used a multitracer stable isotope technique to assess calcium and magnesium balance in 12 boys and 13 girls aged 9-14 y (mean weight: 42 kg) maintained on relatively high calcium intakes (mean: 1310 +/- 82 mg/d). There were no significant differences in absorption of calcium or magnesium from milk between boys and girls. Calcium retention (balance) correlated positively with calcidiol (25-hydroxyvitamin D) concentration (r = 0.48, P = 0.02) and serum alkaline phosphatase activity (r = 0.44, P = 0.03). There was no significant relation between magnesium balance and concentration. When data from this study were combined with our previously reported data, an increase in total calcium absorption was seen for pubertal (Tanner stages 2-4) but not prepubertal (Tanner stage 1) white children over the range of intakes from approximately 750 to 1350 mg/d. Despite intakes similar to the 1989 recommended dietary allowance for magnesium (mean intake: 6.4 +/- 1.2 mg.kg-1.d 1), 11 of the 25 subjects (6 girls and 5 boys) were in negative magnesium balance. We conclude that benefits from higher calcium intakes, < or = 1350 mg/d, were most apparent in pubertal children. In addiction, higher magnesium intakes should be considered for children. PMID- 9356536 TI - Effect of iron supplementation on the iron status of pregnant women: consequences for newborns. AB - We studied the effect of iron supplementation on the iron status of mothers and on biochemical iron status and clinical and anthropometric measures in their infants. The subjects were 197 pregnant women selected at 28 wk +/- 21 d of gestation at a mother-and-child health center in Niamey, Niger. Ninety-nine women received 100 mg elemental Fe/d throughout the remainder of their pregnancies and 98 received placebo. The prevalence of anemia and iron deficiency decreased markedly during the last trimester of pregnancy in the iron-supplemented group but remained constant in the placebo group. Three months after delivery, the prevalence of anemia was significantly higher in the placebo group. At delivery, there were no differences between the two groups in cord blood iron variables. Three months after delivery, serum ferritin concentrations were significantly higher in infants of women in the iron-supplemented group. Mean length and Apgar scores were significantly higher in infants with mothers in the iron group than in those with mothers in the placebo group. PMID- 9356537 TI - Effect of high fiber intake in fish oil-treated patients with non-insulin dependent diabetes mellitus. AB - The short-term effect of high fiber intake on fish-oil treatment in 15 free living, non-insulin-dependent diabetic patients was evaluated by using a controlled, sequential study design. During an 8-wk fish-oil-treatment period when patients received 20 g fish oil/d, the usual daily fiber intake was increased with a 15-g pectin supplement at midpoint. Fish oil alone lowered triacylglycerol and very-low-density-lipoprotein-cholesterol concentrations by 41% and 36%, respectively (both P < 0.01 by the end of the treatment period) with unchanged mean total, low-density-, and high-density-lipoprotein-cholesterol concentrations. When the fiber intake was increased, however, total and low density-lipoprotein-cholesterol concentrations decreased significantly (P < 0.001 and < 0.05, respectively) with fish-oil treatment. The cholesterol ester fraction of plasma lipids was reduced by 34% when compared with fish oil alone (P < 0.05). The plasma triacylglycerol fraction decreased further by 44% (P < 0.001). Other beneficial effects observed included a 30% decline in the fatty acid fraction (P < 0.002) by end of the treatment period. Diabetic control was maintained during the 12-wk study. In conclusion, a high fiber intake may be beneficial in fish oil treated diabetic patients. PMID- 9356538 TI - Increased circulating concentrations of parathyroid hormone in healthy, young women consuming a protein-restricted diet. AB - Increasing dietary protein induces hypercalciuria and a negative calcium balance. Despite this, the influence of dietary protein on the parathyroid hormone (PTH) I a-hydroxylase axis is not well understood. We therefore examined the effect of three amounts of dietary protein: low (0.7 g/kg), medium (1.0 g/kg), and high (2.1 g/kg) on mineral metabolism and the PTH-1-alpha-hydroxylase axis in 16 healthy women aged 26.7 +/- 1.3 y. By day 4, urinary calcium decreased significantly with the low-protein diet and increased significantly with the high protein diet compared with the medium-protein diet (control). Also by day 4, there were striking elevations in serum PTH and calcitriol [1,25-dihydroxyvitamin D] in subjects consuming the low-protein diet. Serum PTH, by two different assays, was 1.5-2.4 times higher and by day 14 1.6-2.7 times higher during the low-protein diet compared with the medium-protein diet. This was accompanied by a significant increase in both nephrogenous cyclic adenosine monophosphate (cAMP), a sensitive and specific indicator of PTH bioactivity, and serum calcitriol by day 14. In comparison, there were relatively minor changes in the calcitropic hormones with the medium- and high-protein diets. The stimulus for the elevation in PTH induced by protein restriction is unclear, but probably does not involve a simple renal mechanism and could reflect either a decline in intestinal calcium absorption, a reduction of bone turn-over, or both. Our data indicate that dietary protein is a powerful regulator of calcium metabolism. Further study is needed to both clarify the mechanisms by which these changes are induced and to better define the amount of dietary protein that will optimize skeletal health in young women. PMID- 9356539 TI - Interactive effects of exercise, alcohol, and vegetarian diet on coronary artery disease risk factors in 9242 runners: the National Runners' Health Study. AB - In a national survey, 199 male and 152 female vegetarian runners and 7054 male and 1837 female omnivorous runners provided data on weekly intakes of alcohol, red meat, fish, and fruit, and weekly distance run. This information was compared with physician-supplied medical data to test whether 1) running benefits vegetarians, 2) alcohol and running distance contribute independently to concentrations of high-density-lipoprotein (HDL) cholesterol, and 3) running mitigates the hypertensive effects of alcohol. Greater reported weekly distance run by vegetarians was associated with greater HDL-cholesterol concentrations [slopes +/- SEs for men and women, respectively: 0.003 +/- 0.001 and 0.005 +/- 0.002 (mmol/L)/km] and lower waist (-0.06 +/- 0.02 and-0.08 +/- 0.02 cm/km), hip (-0.05 +/- 0.03 and -0.07 +/- 0.02 cm/km), and chest (-0.05 +/- 0.02 cm/km for both) circumferences. In men and women, alcohol and running distance contributed independently to higher HDL-cholesterol concentrations. Men who ran > 72 km and drank > 177 mL (6 oz) alcohol/wk were five times more likely to have clinically defined high HDL cholesterol (> or = 1.55 mmol/L, or > or = 60 mg/dL) than were nondrinkers running < 24 km/wk. Regardless of running level, men's blood pressure increased in association with alcohol intake. These data suggest that 1) running distance in vegetarians and vegans has the same relation to HDL cholesterol (increasing) and adiposity (decreasing) as reported previously for omnivores, 2) alcohol and running distance contribute independently to higher HDL cholesterol, and 3) running does not abate the hypertensive effects of alcohol in men. Also, vigorous exercise provides important health benefits beyond those obtained by diet. PMID- 9356540 TI - Effects of family history of heart disease, apolipoprotein E phenotype, and lipoprotein(a) on the response of children's plasma lipids to change in dietary lipids. AB - We examined the effects of family history of coronary artery disease (CAD), apolipoprotein E (apo E) phenotype, and lipoprotein(a) [Lp(a)] on the response of plasma lipids to change in dietary lipid intake after 3 mo of nutrition education in 125 children aged 4-10 y. The subjects were healthy children with elevated low density-lipoprotein (LDL)-cholesterol concentrations who participated in the Children's Health Project, a nutrition-education program designed to lower plasma cholesterol by means of dietary modifications in accordance with recommendations of the National Cholesterol Education Program. Dietary and plasma lipids were measured by three 24-h recalls and assessments of two fasting plasma samples collected before and 3 mo after the start of intervention. Family history of CAD was determined by questionnaires administered to parents at baseline. Apo E phenotyping was done with isoelectric focusing followed by immunostaining; Lp(a) was measured with two-site immunoradiometric assays of frozen aliquots of plasma samples collected at baseline and 3 mo. After adjustment for intervention group, age, sex, and body mass index, analysis of covariance showed that baseline plasma lipid concentrations were the strongest independent predictors of change in plasma lipids after 3 mo. Plasma total and LDL-cholesterol concentrations in children with less family history of CAD were significantly more responsive to change in dietary cholesterol than concentrations in children with a stronger family history of CAD. Neither apo E phenotype nor Lp(a) significantly influenced change in plasma lipids independently or interactively with change in dietary lipids. PMID- 9356541 TI - Effect of vitamin E supplementation on prostaglandin concentrations in aspirin induced acute gastric injury in aged rats. AB - Nonsteroidal antiinflammatory drugs (NSAIDs), such as aspirin, frequently cause gastric mucosal injury in the elderly. Impairment of prostaglandin synthesis is a crucial step by which aspirin attenuates mucosal defense capacity. Vitamin E has been shown to decrease prostanoid concentrations, which implies an ulceropermissive effect of vitamin E. To assess the effect of vitamin E on aspirin-induced gastric injury and mucosal prostanoid concentrations, 20 male rats aged 20 mo were divided into two groups and fed diets containing either 30 (physiologic requirement) or 500 mg all-rac-alpha-tocopheryl acetate/kg. After 6 wk, all rats received two intragastric doses of aspirin (1.4 mumol/kg body wt). A third group of six animals fed the high-vitamin E diet received a vehicle solution without aspirin. Mucosal samples for vitamin E and prostaglandin E2, 6 keto-prostaglandin F1 alpha, and thromboxane A2 measurements were collected. The prevalence and degree of mucosal lesions were not significantly different among all groups. Rats fed the high-vitamin E diet had significantly higher mucosal vitamin E concentrations than rats fed the low-vitamin E diet. Mucosal concentrations of all three prostanoids were 95% lower in aspirin-treated rats than in controls (P = 0.0001 in all instances). The high-vitamin E diet group had significantly lower mucosal 6-keto-prostaglandin F1 alpha concentrations (P = 0.02) than the low-vitamin E diet group, indicating decreased prostacyclin formation, whereas concentrations of prostaglandin E2 and thromboxane A2 were similar in the aspirin-treated groups. Aspirin markedly reduced mucosal prostanoid concentrations in rats, without apparent effects on gastric injury, whereas vitamin E supplementation significantly reduced mucosal 6-keto prostaglandin F(1 alpha) concentrations. Nevertheless, vitamin E supplementation did not result in more gastric injury in aspirin-treated rats than in controls. PMID- 9356543 TI - Prevalence of malnutrition in nonsurgical hospitalized patients and its association with disease complications. AB - The prevalence of malnutrition and its predictive value for the incidence of complications were determined in 155 patients hospitalized for internal or gastrointestinal diseases. At admission, 45% of the patients were malnourished according to the Subjective Global Assessment (physical examination plus questionnaire), 57% according to the Nutritional Risk Index [(1.5 x albumin) + (41.7 x present/usual weight)], and 62% according to the Maastricht Index [(20.68 - (0.24 x albumin) - (19.21 x transthyretin (prealbumin) - (1.86 x lymphocytes) - (0 04 x ideal weight)]. Crude odds ratios for the incidence of any complication in malnourished compared with well-nourished patients during hospitalization were 2.7 (95% CI: 1.4, 5.3) for the Subjective Global Assessment, 2.8 (1.5, 5.5) for the Nutritional Risk Index, and 3.1 (1.5, 6.4) for the Maastricht Index. Odds ratios were reduced to 1.7 (0.8, 3.6), 1.6 (0.7, 3.3), and 2.4 (1.1, 5.4), respectively, after a multivariate analysis that included disease category and disease severity. Because the confounding factors adjusted for are not only a measure of the severity of the disease but may also be influenced by malnutrition itself, the actual risk for complications due to malnutrition could be higher than the adjusted odds ratios. In conclusion, malnutrition was frequent in patients with gastrointestinal disease and other internal diseases at the time of admission. The severity of malnutrition in the patients predicted the occurrence of complications during their hospital stay and this association was not completely explained by confounding factors. PMID- 9356542 TI - Insulin sensitivity and intake of vitamins E and C in African American, Hispanic, and non-Hispanic white men and women: the Insulin Resistance and Atherosclerosis Study (IRAS). AB - Elevated fasting insulin concentrations and insulin resistance have been associated with non-insulin-dependent diabetes mellitus (NIDDM), obesity, atherosclerosis, and hypertension. Vitamin E supplementation in persons with and without NIDDM may be related to greater insulin sensitivity (SI). The cross sectional associations of the intake of vitamins E and C with SI and insulin concentrations were evaluated among African American, Hispanic, and non-Hispanic white men and women with a wide spectrum of glucose tolerance included in the Insulin Resistance and Atherosclerosis Study (IRAS) (n = 1151). Insulin sensitivity was measured by minimal model analysis of a 12-sample, insulin modified, frequently sampled intravenous glucose tolerance test. Nutrient intake (including vitamin supplement use) was assessed with a food-frequency questionnaire modified to include foods consumed by the three ethnic groups. Linear-regression models were used, including rank of SI and the log of fasting insulin as the outcome variables. Pearson correlation coefficients for vitamins E and C in relation to rank SI were r = 0.07 (P = 0.01) and r = 0.07 (P = 0.02), respectively. After adjustment for total energy and BMI these associations were no longer statistically significant and did not differ between ethnic groups. Results were similar when vitamins E and C were combined in categories of low and high antioxidant intake. Models replicated with log of fasting insulin as the outcome variable also did not produce significant associations with vitamins E or C. Thus, these cross-sectional analyses do not support the hypothesis of improved SI with increased intake of vitamins E and C. PMID- 9356544 TI - Effect of cholesterol-rich diets with and without added vitamins E and C on the severity of atherosclerosis in rabbits. AB - Oxysterols as oxidation products of cholesterol are considered an atherogenic factor in the development of atherosclerosis in the arteries of cholesterol-fed rabbits. We compared the atherogenic effects of diets enriched either with 0.5% oxidized cholesterol (OC; characterized by high amounts of oxysterols) or with pure cholesterol (PC). The effects of antioxidant vitamins E and C added to the PC diet were also evaluated in view of their antioxidative properties for lipoproteins and cholesterol and how this could affect the severity of atherosclerosis. Four groups of rabbits were fed the following for 11 wk: 1) a nonpurified stock diet, 2) this stock diet plus 0.5% OC, 3) the stock diet plus 0.5% PC, and 4) the stock diet plus 0.5% PC and 1000 mg vitamin E and 500 mg vitamin C/kg diet (PC + antioxidants). The OC and PC diets were equally hyperlipidemic and hypercholesterolemic. The severity of atherosclerotic lesions was highest with the OC diet and lowest with the PC + antioxidants diet. The plasma oxysterol concentration was proportional to the severity of atherosclerosis in all three groups of cholesterol-fed rabbits. beta-Very-low density-lipoprotein modification was minimized by vitamins E and C as indicated by its polyacrylamide gel electrophoretic pattern and its increased binding to the rabbit liver membrane in vitro. This study indicated that OC and PC were equally atherogenic but that the addition of antioxidants to the PC diet significantly reduced its severity, even when hypercholesterolemia persisted. This indicated that atherogenesis can result from an excessive accumulation of oxidation products of cholesterol in the plasma. PMID- 9356545 TI - Predicting energy needs in ventilator-dependent critically ill patients: effect of adjusting weight for edema or adiposity. AB - Predicting energy needs in critical illness can be difficult because of uncertainties about the influence of multiple factors on energy expenditure. Understanding these components is important to avoid limiting optimal outcome by underfeeding and to avoid complications of overfeeding. Prediction strategies often use a patient's weight to estimate needs. For overweight patients, there is controversy as to whether actual or modified weight should be used in predictions. This study was designed to evaluate a proposed technique to improve the accuracy of predicting energy needs in critically ill, overweight subjects. Subjects' energy needs were predicted [with Harris-Benedict equation (HBE) and kilojoules per kilogram (KPK) strategies] by using both actual weight and an adjusted weight developed to attempt to more accurately reflect lean mass. Results were compared with measured energy expenditure determined by indirect calorimetry. Results indicated that use of actual weights in predictions for overweight subjects may lead to overfeeding. Use of adjusted weights led to more accurate energy predictions with the KPK than with the HBE strategy. Adjusted weight strategies could explain > 45% of the variability of resting energy expenditure in subjects 130-159% of ideal body weight. Results of this study suggest that using adjusted weights with the KPK prediction strategy may be preferable for this population, particularly for patients > or = 130% of ideal body weight. This study also indicated that multiple diagnoses may not lead to increased energy requirements. PMID- 9356546 TI - A cross-sectional study of dental caries, intake of confectionery and foods rich in starch and sugars, and salivary counts of Streptococcus mutans in children in Spain. AB - In this cross-sectional study of 236 schoolchildren living in Manresa, Spain, we evaluated the association between prevalence of dental caries and frequency of consumption of various food groups, including sweetened baked goods and similar foods (rich in starch and sugars) and confectionery (rich in sugars but not starch), using a food-frequency questionnaire. Because Streptococcus mutans is associated with the cariogenicity of carbohydrates, we also evaluated the modification of these associations by salivary counts of this microorganism. Odds ratios (ORs) were used to measure the association between caries and tertiles of consumption. Sex, age, use of fluorides, tooth-brushing frequency, frequency of dental visits, socioeconomic status, and intake of other potentially cariogenic food groups were considered as potential confounders. We did not find a significant association between any of the food groups evaluated and caries prevalence. Failure to detect an association could have been due to the low prevalence of caries in our population (decayed, missing, or filled permanent teeth = 1.3 at age 10.6 y) or to underestimation of the association due to diet misclassification. In this population, the association between consumption of sweetened baked goods and caries appeared to be modified by the numbers of S. mutans [OR = 6.1 (95% CI: 1.6, 23.0) for low compared with high intake in children with moderate-to-high S. mutans counts and OR = 0.3 (95% CI: 0.1, 1.6) for low compared with high intake in children with low S. mutans counts]. These results suggest that a high intake of sweetened baked goods may be a determinant of caries prevalence in children with moderate-to-high salivary counts of S. mutans. PMID- 9356547 TI - An insulin index of foods: the insulin demand generated by 1000-kJ portions of common foods. AB - The aim of this study was to systematically compare postprandial insulin responses to isoenergetic 1000-kJ (240-kcal) portions of several common foods. Correlations with nutrient content were determined. Thirty-eight foods separated into six food categories (fruit, bakery products, snacks, carbohydrate-rich foods, protein-rich foods, and breakfast cereals) were fed to groups of 11-13 healthy subjects. Finger-prick blood samples were obtained every 15 min over 120 min. An insulin score was calculated from the area under the insulin response curve for each food with use of white bread as the reference food (score = 100%). Significant differences in insulin score were found both within and among the food categories and also among foods containing a similar amount of carbohydrate. Overall, glucose and insulin scores were highly correlated (r = 0.70, P < 0.001, n = 38). However, protein-rich foods and bakery products (rich in fat and refined carbohydrate) elicited insulin responses that were disproportionately higher than their glycemic responses. Total carbohydrate (r = 0.39, P < 0.05, n = 36) and sugar (r = 0.36, P < 0.05, n = 36) contents were positively related to the mean insulin scores, whereas fat (r = -0.27, NS, n = 36) and protein (r = -0.24, NS, n = 38) contents were negatively related. Consideration of insulin scores may be relevant to the dietary management and pathogenesis of non-insulin-dependent diabetes mellitus and hyperlipidemia and may help increase the accuracy of estimating preprandial insulin requirements. PMID- 9356549 TI - Effect of inflammation on measures of antioxidant status in patients with non small cell lung cancer. AB - This study examined the effect of an inflammatory response on measures of antioxidant status in patients with non-small cell lung cancer (NSCLC). In healthy, control subjects (n = 13) and NSCLC patients (n = 22) fasting concentrations of albumin, C-reactive protein, cholesterol, and the antioxidants alpha-tocopherol, retinol, lutein, lycopene, and alpha- and beta-carotene were measured. The two groups were similar in terms of age, sex, and body mass index. However, the cancer group had an inflammatory response as evidenced by significantly increased C-reactive protein concentrations. Concentrations of all the measured antioxidants of the NSCLC group were significantly lower than those of the control group (P < 0.01). The lowest concentrations were those of the carotenoids lycopene and alpha- and beta-carotene. In the cancer group there were significant negative correlations between concentrations of C-reactive protein and retinol (r = -0.682, P < 0.01), alpha-tocopherol (r = -0.464, P < 0.05), and lutein (r = -0.599, P < 0.01). The results of this study have implications for the interpretation of circulating antioxidant concentrations in patients with NSCLC. PMID- 9356548 TI - Shift from a dairy product-rich to a dairy product-free diet: influence on cytotoxicity and genotoxicity of fecal water--potential risk factors for colon cancer. AB - Several epidemiologic studies have suggested that dairy product intake is associated with a decreased incidence of colon cancer. To determine whether the cytotoxicity and genotoxicity of the aqueous portion of human stool (two potential risk markers for the disease) were affected by a change in dairy product intake, 18 healthy male and female volunteers were randomly divided into two groups. In a crossover design, the volunteers shifted from their normal dairy product-rich diet to a dairy product-free diet. Nutritional analysis of the food consumed during the study period showed a significant decrease in energy intake from 9000 to 7866 kJ/d because of a decreased intake of protein and fat. Carbohydrate and fiber intakes remained unchanged during the intervention. Calcium intake decreased significantly from 1488 to 372 mg/d, with similar significant decreases in phosphate and vitamin D intakes. Cytotoxicity of fecal water, analyzed by the HT-29 cytotoxicity assay, indicated a significant decrease in cell survival from 34% to 20% when dairy products were excluded from the participants' diets. Single-cell gel electrophoresis (COMET assay), used to analyze genotoxicity of fecal waters, indicated no differences brought about by the dietary intervention. In conclusion, our findings indicate that a shift from a dairy product-rich to a dairy product-free diet resulted in a significant effect on an accepted risk marker for colon cancer and may suggest that the mechanism by which dairy products are protective is at the level of tumor promotion rather than initiation. PMID- 9356551 TI - Low-fat, high-carbohydrate diets and risk factors for ischemic heart disease. PMID- 9356550 TI - Bacterial fermentation of fructooligosaccharides and resistant starch in patients with an ileal pouch-anal anastomosis. AB - Patients with large bowel disease may undergo ileal pouch-anal anastomosis, in which the colon is removed and part of the distal ileum is used to construct a pelvic reservoir. Competence of the ileal pouch to ferment carbohydrates is associated with the absence of pouchitis. However, the extent to which bacterial fermentation takes place and whether it is affected by diet are unclear. We investigated fermentation of two nondigestible carbohydrates, fructooligosaccharides and resistant starch, in 15 healthy patients with an ileal pouch by using a placebo-controlled crossover design (with glucose as the placebo). Apparent fermentability of fructooligosaccharides was 83%; that of resistant starch was 46%. Resistant starch increased fecal excretion of butyrate by 69% whereas fructooligosaccharides reduced excretion of amino acid-derived isobutyrate by 94% and of isovalerate by 77%. Fructooligosaccharides also significantly increased fecal weight (651 compared with 541 g/d) and breath hydrogen excretion (286 compared with 85 ppm x h). Bacterial fermentation of nondigestible carbohydrates in pouches takes place to an appreciable extent and in a substrate-specific manner. The relation between such fermentation and inflammation of the pouch (pouchitis) deserves study. PMID- 9356552 TI - Body mass index in patients with unusual proportions. PMID- 9356553 TI - WHO anthropometric reference data, with special reference to growth in adolescence. PMID- 9356554 TI - We can talk: individual psychotherapy for schizophrenia. PMID- 9356555 TI - Images in neuroscience. Clinical genetics, V. Association of genetic and personality characteristics. PMID- 9356556 TI - DST studies in psychotic depression: a meta-analysis. AB - OBJECTIVE: Although hypersecretion of cortisol has frequently been reported in psychotic depression, the findings have been mixed. The authors performed a meta analysis of the existing studies to determine the significance of differences in nonsuppression of cortisol across studies. METHOD: Fourteen studies that compared dexamethasone suppression test (DST) results in psychotic and nonpsychotic patients were examined, and a Mantel-Haenszel meta-analysis was performed. For comparison purposes, 19 studies of the DST with respect to melancholic/nonmelancholic and inpatient/outpatient distinctions were similarly reviewed. RESULTS: This analysis indicated a highly significant probability that a greater rate of cortisol nonsuppression occurs in psychotic depression. The nonsuppression rate was substantially higher in patients with psychotic depression (64%) than in nonpsychotic patients (41%). In the 19 studies of melancholia, nonsuppression was less frequent (36%), and when inpatient/outpatient status was controlled, melancholic depression was not significantly associated with nonsuppression. In nonmelancholic outpatients with major depression, the nonsuppression rate was low (12%). CONCLUSIONS: Among patients with major depression, psychotic depression is the subtype that is most closely associated with nonsuppression of cortisol on the DST. Hypercortisolemia is usually present in psychotic depression and may be important in understanding the pathophysiology of this syndrome. PMID- 9356557 TI - Three-year trials of personal therapy among schizophrenic patients living with or independent of family, I: Description of study and effects on relapse rates. AB - OBJECTIVE: The study of individual psychotherapeutic approaches to the treatment of schizophrenia has yielded equivocal findings, partly because of methodologic problems. Further, the ability of psychosocial treatments to prevent psychotic relapse appears to lessen over time. The authors' goal was to develop and test a demonstrably effective individual therapy for schizophrenia. METHOD: Using a study design that addressed previous methodologic issues, the authors evaluated personal therapy specifically designed to forestall late relapse in patients with schizophrenia. They evaluated the effectiveness of personal therapy over a period of 3 years after hospital discharge among 151 patients with schizophrenia or schizoaffective disorder diagnosed according to Research Diagnostic Criteria. The patients were randomly assigned to receive either personal therapy or contrasting therapies in one of two concurrent trials. One trial studied patients who were living with family (N = 97); the other studied patients who were living independent of family (N = 54). RESULTS: All of the patients had extensive psychiatric histories, but only 44 (29%) experienced recurrent psychotic episodes over the 3-year study period, and only 27 (18%) prematurely terminated the study; most of those who left the study were in the no-personal-therapy conditions. Among patients living with family, personal therapy was more effective than family and supportive therapies in preventing psychotic and affective relapse as well as noncompliance. However, among patients living independent of family, those who received personal therapy had significantly more psychotic decompensations than did those who received supportive therapy. CONCLUSIONS: Personal therapy had a positive effect on adverse outcomes among patients who lived with family. However, personal therapy increased the rate of psychotic relapse for patients living independent of family. The application of personal therapy might best be delayed until patients have achieved symptom and residential stability. PMID- 9356558 TI - Three-year trials of personal therapy among schizophrenic patients living with or independent of family, II: Effects on adjustment of patients. AB - OBJECTIVE: Previous analyses of the personal and social adjustment of outpatients with schizophrenia have either relied on the assessment of unrepresentative patients who survived without relapse or used analyses that included relapse assessments, a potential confound when different rates of relapse existed among treatment conditions. The authors' goal was to conduct a study of the effects of personal therapy on outcome that was designed to take into consideration the effects of relapse. METHOD: They evaluated the effectiveness of personal therapy over 3 years after hospital discharge among 151 patients with schizophrenia or schizoaffective disorder. The patients were randomly assigned to receive personal therapy or contrasting therapies in one of two concurrent trials. One trial included patients who were living with family (N = 97); the other included patients who were living independent of family (N = 54). Patients were assessed at 6-month intervals over 3 years of treatment on measures of personal and social adjustment; patients who relapsed and restabilized and those who did not relapse were included. RESULTS: Personal therapy had positive effects on broad components of social adjustment (role performance) but had few differential effects on symptoms, and patients receiving personal therapy remained more anxious than patients who received family or supportive therapy. For patients who were living with family, personal therapy led to better outcomes in overall performance than did the other treatments. Although family therapy had only one positive effect on patients' social adjustment, the personal adjustment (residual symptoms) of patients who received family therapy appeared to improve more than that of patients receiving personal or supportive therapy. For patients not living with family, personal therapy was more successful than supportive therapy in improving work performance and relationships out of the home. Longitudinal effects of personal therapy on symptoms were similar to those of family and supportive therapies, particularly in the first 2 years, but personal therapy effect sizes increased over time on measures of social adjustment. CONCLUSIONS: Personal therapy has pervasive effects on the social adjustment of patients with schizophrenia that are independent of relapse prevention. Supportive therapy, with or without family intervention, produces adjustment effects that peak at 12 months after discharge and plateau thereafter. However, personal therapy, a definitive psychosocial intervention, continues to improve the social adjustment of patients in the second and third years after discharge. Brief treatment would appear to be less effective than a long-term, disorder-relevant intervention for schizophrenia. PMID- 9356560 TI - Clinical and neurocognitive aspects of source monitoring errors in schizophrenia. AB - OBJECTIVE: Source monitoring, an aspect of memory that involves judgments about the origin of information, has been found to be more prone to errors in schizophrenic subjects than in normal persons. To examine the precise nature of such errors and their relationship to clinical and neurocognitive variables, the authors compared schizophrenic and normal subjects. METHOD: Schizophrenic subjects who had been medication free for 1 week (N = 26) and demographically matched normal subjects (N = 21) performed a source monitoring task and were assessed on current psychiatric symptoms, IQ, and frontal lobe functioning. RESULTS: The schizophrenic subjects had normal recognition memory of target words (recognition hits) and a normal generation effect but made more errors than the comparison subjects in identifying the source of target words. Specifically, the schizophrenic subjects made more errors in remembering the source of new and self generated items, and they tended to attribute items to an external source. In 11 retested subjects, these errors were stable and independent from medication status after a 2-year interval. Secondary analyses suggested that certain source monitoring errors may be associated with hostility and lower IQ. When the effect of IQ was controlled, correlations with frontal dysfunction were not significant. CONCLUSIONS: Schizophrenic subjects make significantly more source monitoring errors than normal subjects, but not because of problems with recognition memory hits or with the generation effect. This tendency may be trait like and may be related to hostility. Lower IQ in schizophrenia plays a partial role in these errors, but frontal dysfunction does not. PMID- 9356559 TI - PET evidence that loxapine is an equipotent blocker of 5-HT2 and D2 receptors: implications for the therapeutics of schizophrenia. AB - OBJECTIVE: Loxapine, a dibenzoxazepine antipsychotic, is closely related to clozapine and shares clozapine's high affinity for binding to serotonin 5-HT2 and dopamine D4 receptors. The purpose of this study was to document loxapine's 5-HT2 and D2 receptor occupancy in vivo in patients with psychoses. METHOD: Ten patients who were taking loxapine (10-100 mg/day) had their D2 and 5-HT2 receptors assessed by means of positron emission tomography with [11C]raclopride and [18F]setoperone, respectively. RESULTS: The D2 receptor occupancy ranged from 43% to 90%; 5-HT2 occupancy varied from 27% to near saturation. Statistical comparison of the results showed that loxapine was equipotent in blocking 5-HT2 and D2 receptors. CONCLUSIONS: Loxapine differs from typical neuroleptics in demonstrating a high degree of 5-HT2 receptor occupancy. However, it is not "atypical" like clozapine and risperidone, since its 5-HT2 occupancy is not higher than its D2 occupancy. The results demonstrate that a high level of 5-HT2 occupancy is not a sufficient condition for atypicality. If atypical antipsychotic action is predicated on a combination of 5-HT2 and D2 effects, then it requires > 80% 5-HT2 occupancy in conjunction with < 80% D2 occupancy. PMID- 9356561 TI - Clinical correlates of memory in schizophrenia: differential links between depression, positive and negative symptoms, and two types of memory impairment. AB - OBJECTIVE: This study investigated clinical correlates of memory impairment in schizophrenic patients. In particular, the authors hypothesized that depressive symptoms would be linked to memory efficiency, as found in other clinical populations. In addition, they tested Frith's pathophysiological model predicting links between negative symptoms and failure to respond, as well as between positive symptoms and production of erroneous responses. METHOD: Thirty-one patients were given several memory tasks: long-term free recall of nonorganizable and organizable lists in immediate and delayed conditions, recognition in immediate and delayed conditions, implicit memory (stem completion task), and short-term memory (digit span). Superficial encoding of information was also assessed by the ability to recall the items sequentially; deep encoding was assessed by the ability to organize the items according to their semantic properties. Two types of memory measures were individualized: measures reflecting memory efficiency and measures reflecting production of erroneous memory responses (intrusions, perseverations, false alarms). RESULTS: Consistent correlations appeared between severity of depressive symptoms and measures reflecting deep but not superficial encoding; none, however, was correlated with negative symptoms. Two of the three types of erroneous memory responses were positively linked to positive symptoms. CONCLUSIONS: Efficiency of memory processes relying on deep encoding seemed linked to depressive symptoms. In addition, the two distinct types of impairment predicted by Frith's model were found. The expected link of one with positive symptoms was verified, but the link of the other with negative symptoms was not. PMID- 9356562 TI - Premorbid social functioning in schizophrenia and bipolar disorder: similarities and differences. AB - OBJECTIVE: This research examined social functioning in childhood and adolescence among patients with schizophrenia and patients with bipolar disorder compared with healthy subjects and investigated the relation between premorbid adjustment and risk factors for psychosis. METHOD: Maternal recall was used to assess the premorbid adjustment of patients with schizophrenia (N = 70) and patients with bipolar disorder (N = 28) recruited from a survey of consecutive hospital admissions for psychosis and of healthy comparison subjects (N = 100) drawn from the same catchment area. RESULTS: The patients with schizophrenia had significantly poorer premorbid adjustment in childhood and adolescence than the comparison subjects and were impaired in both sociability and school adjustment. The patients with bipolar disorder exhibited poorer social impairment in adolescence than the comparison subjects, though to a lesser degree than the schizophrenic subjects, but functioned well at school. There were significant linear trends in the risk of psychosis with worsening premorbid functioning, which was most marked in the schizophrenic group, and a specific linear relation between low birth weight and poor premorbid adjustment among the schizophrenic patients. CONCLUSIONS: Impaired premorbid social functioning is not specific to schizophrenia and is seen also in bipolar disorder. The data support the view that poor premorbid social adjustment is one manifestation of vulnerability to adult psychotic disorders. These results are consistent with other findings pointing to early developmental deficits in patients who subsequently develop psychosis. PMID- 9356563 TI - Depressive symptoms in the early course of schizophrenia: relationship to familial psychiatric illness. AB - OBJECTIVE: This study examined the relation between the presence of depressive symptoms in schizophrenic patients with a recent first psychotic episode and affective disorders among their relatives. METHOD: Data on depressive symptoms in 70 patients with schizophrenia diagnosed according to the DSM-III-R criteria, who had had a recent first psychotic episode, and psychiatric diagnostic information on 293 of their first-degree and 674 of their second-degree relatives were collected. Depressive symptoms in the schizophrenic probands were examined at the index psychotic episode (at study entry) and systematically over a 1-year follow through period. The majority of first-degree family members were interviewed in person with the use of semistructured diagnostic interviews. RESULTS: The linear regression findings confirmed the hypothesis that depressive symptoms in the early course of schizophrenia are associated with a family history of unipolar affective illness. CONCLUSIONS: Because depression in the patients was associated with a family history of depression, this suggests that depression in schizophrenia is not solely either a reaction to having had a psychotic episode or part of the recovery process. The findings are consistent with a model in which a familial genetic liability to affective disorder, when present, is viewed a s exerting a modifying influence on the patient's schizophrenic illness to increase expression of depressive symptoms. PMID- 9356564 TI - Respiratory psychophysiology of panic disorder: three respiratory challenges in 98 subjects. AB - OBJECTIVE: Respiratory abnormalities may play a central role in the pathophysiology of panic disorder. The current study was undertaken to examine the respiratory response in the largest series of subjects to date during three respiratory challenges that used improved methodology. METHOD: Fifty-nine patients with DSM-III-R panic disorder and 39 normal volunteers were challenged with 5% and 7% CO2 inhalation and room air hyperventilation separated by room air breathing with continuous spirometry. RESULTS: Patients with panic disorder were more sensitive to the anxiogenic effects of CO2 than were normal subjects, and CO2 was a more potent stimulus to panic than hyperventilation. Patients increased their respiratory rate more quickly during CO2 inhalation than did comparison subjects, and this increase preceded the panic attacks. Patients who panicked in response to 5% CO2 demonstrated continued rise in end-tidal CO2, while the end tidal CO2 of the comparison groups stabilized. Low end-tidal CO2 and high variance in minute ventilation at baseline predicted panic attacks during CO2 inhalation. Following CO2 or hyperventilation challenges, respiratory rate dropped sharply, while tidal volume remained elevated longer in patients than in comparison subjects. CONCLUSIONS: The findings confirm the greater behavioral and physiological sensitivity of patients with panic disorder to CO2 inhalation and identify a series of respiratory abnormalities. Panic attacks in panic disorder may be explained by inefficient compensatory mechanisms, primarily of respiratory rate. PMID- 9356565 TI - Association of panic disorder with a history of traumatic suffocation. AB - OBJECTIVE: An important recent hypothesis suggests that panic disorder results from a false suffocation alarm. However, the association of panic disorder with a history of traumatic suffocation experiences (e.g., near-drowning and near choking) has not been well studied. This study examined whether a history of traumatic suffocation might be more common in patients with panic disorder who have predominantly respiratory symptoms. METHOD: Patients with panic disorder (N = 176) and psychiatric comparison subjects (N = 60) were questioned about a history of traumatic suffocation experiences. The panic disorder patients were classified as having predominantly respiratory, cardiovascular, occulovestibular, or gastrointestinal symptoms in order to determine a possible association between traumatic suffocation and symptom subtype. RESULTS: The frequency of traumatic suffocation was significantly higher among the panic disorder patients (19.3%) than among the comparison subjects (6.7%). Within the panic disorder group, patients with a history of traumatic suffocation were significantly more likely to exhibit predominantly respiratory symptoms and nocturnal panic attacks, while patients without such a history were significantly more likely to have predominantly cardiovascular symptoms, occulovestibular symptoms, and agoraphobia. CONCLUSIONS: There may be a specific association between panic disorder and a history of traumatic suffocation, and such a history in turn appears associated with predominantly respiratory symptoms and nocturnal panic attacks. Although additional studies are needed to confirm these data, a history of traumatic suffocation might be hypothesized to play a role in the etiology of panic disorder in some patients and may provide a useful window on understanding the psychobiology of this disorder. PMID- 9356566 TI - Multicenter collaborative panic disorder severity scale. AB - OBJECTIVE: To address the lack of a simple and standardized instrument to assess overall panic disorder severity, the authors developed a scale for the measurement of panic disorder severity. METHOD: Ten independent evaluators used the seven-item Panic Disorder Severity Scale to assess 186 patients with principal DSM-III-R diagnoses of panic disorder (with no or mild agoraphobia) who were participating in the Multicenter Collaborative Treatment Study of Panic Disorder. In addition, 89 of these patients were reevaluated with the same scale after short-term treatment. A subset of 24 patients underwent two independent assessments to establish interrater reliability. Internal consistency, convergent and discriminant validity, and sensitivity to change were also determined. RESULTS: The Panic Disorder Severity Scale was associated with excellent interrater reliability, moderate internal consistency, and favorable levels of validity and sensitivity to change. Individual items showed good convergent and discriminant validity. Analysis suggested a two-factor model fit the data best. CONCLUSIONS: The Panic Disorder Severity Scale is a simple, efficient way for clinicians to rate severity in patients with established diagnoses of panic disorder. However, further research with more diverse groups of panic disorder patients and with a broader range of convergent and discriminant validity measures is needed. PMID- 9356567 TI - Posttraumatic stress disorder in a community group of former prisoners of war: a normative response to severe trauma. AB - OBJECTIVE: The goal of this study was to assess and describe the long-term impact of traumatic prisoner of war (POW) experiences within the context of posttraumatic psychopathology. Specifically, the authors attempted to investigate the relative degree of normative response represented by posttraumatic stress disorder (PTSD) in comparison to other DSM axis I disorders often found to be present, either alone or concomitant with other disorders, in survivors of trauma. METHOD: A community group of 262 U.S. World War II and Korean War former POWs was recruited. These men had been exposed to the multiple traumas of combat, capture, and imprisonment, yet few had ever sought mental health treatment. They were assessed for psychopathology with diagnostic interviews and psychodiagnostic testing. Regression analyses were used to assess the contributions of age at capture, war trauma, and postwar social support to PTSD and the other diagnosed disorders. RESULTS: More than half of the men (53%) met criteria for lifetime PTSD, and 29% met criteria for current PTSD. The most severely traumatized group (POWs held by the Japanese) had PTSD lifetime rates of 84% and current rates of 59%. Fifty-five percent of those with current PTSD were free from the other current axis I disorders (uncomplicated PTSD). In addition, 34% of those with lifetime PTSD had PTSD as their only lifetime axis I diagnosis. Regression analyses indicated that age at capture, severity of exposure to trauma, and postmilitary social support were moderately predictive of PTSD and only weakly predictive of other disorders. CONCLUSIONS: These findings indicate that PTSD is a persistent, normative, and primary consequence of exposure to severe trauma. PMID- 9356568 TI - Prevalence and characteristics of trauma and posttraumatic stress disorder in a southwestern American Indian community. AB - OBJECTIVE: High rates of violence and trauma in many American Indian communities have been reported. The authors investigated the relationship between both the frequency and type of traumatic events and the prevalence of posttraumatic stress disorder (PTSD) in a Southwestern American Indian tribe. METHOD: A structured psychiatric interview and the Traumatic Events Booklet were administered to a subset of 247 tribal members from an overall study population of 582. Subjects were recruited from the community on the basis of membership in pedigrees, and not by convenience. DSM-III-R diagnoses were assigned by consensus after the interviews were evaluated blindly by independent raters. RESULTS: The prevalence of lifetime PTSD was 21.9% (N = 54), and 81.4% of the subjects (N = 201) had experienced at least one traumatic event apiece. The most predictive factor for lifetime PTSD among women was the experience of physical assault, and for men the most predictive factors were a history of combat and having experienced more than 10 traumatic events. CONCLUSIONS: In this Southwestern American Indian community, the prevalences of lifetime PTSD and of exposure to a traumatic event were higher than in the general U.S. population. However, the nearly 4:1 ratio of subjects who reported at least one traumatic event to those with PTSD diagnoses is similar to findings from studies of non-Indians. Individuals with a history of multiple traumatic events (66.0%, N = 163) had a significantly higher risk of developing PTSD. Chronic and multiple trauma did not preclude the identification of acute and discrete traumatic events that resulted in PTSD. PMID- 9356569 TI - What happens to anxiety levels on giving up smoking? AB - OBJECTIVE: DSM-IV lists increased anxiety as a nicotine withdrawal symptom. Increased anxiety has been reported to follow smoking cessation in most but not all studies. Indeed, there is some evidence for a reduction in anxiety, compared with precessation levels, after the first few weeks of abstinence. This study reports data from 101 smokers who attempted to stop smoking and who satisfied DSM III-R criteria for nicotine dependence. METHOD: Unlike most studies in this area, a strict criterion of lapse-free abstinence was adopted. It is argued that lapses during an attempt at cessation may underlie a transient increase in anxiety. Anxiety was measured both by a single rating typical in withdrawal studies and by the State-Trait Anxiety Inventory--State Form X. Patients were rated 2 weeks and 1 week before cessation, immediately before cessation, 24 hours after cessation, and 1, 2, 3, and 4 weeks after cessation. RESULTS: Seventy patients were abstinent for the 4-week follow-up period. There was no evidence of an increase in anxiety following smoking cessation. However, there was a significant decrease in anxiety from the first week of abstinence. CONCLUSIONS: The results weaken the view that increased anxiety is a robust and central element of the nicotine withdrawal syndrome and suggest that giving up smoking is quite rapidly followed by a reduction in anxiety that may reflect removal of an anxiogenic agent, nicotine. PMID- 9356570 TI - Comparability of telephone and face-to-face interviews in assessing axis I and II disorders. AB - OBJECTIVE: The present study examined the comparability of data obtained by telephone and face-to-face interviews for diagnosing axis I and II disorders. METHOD: Sixty young adults from the community were interviewed face-to-face and over the telephone regarding axis I disorders; another 60 subjects were interviewed twice regarding axis II disorders. The order of interviews was counterbalanced, and subjects with a history of disorder were oversampled. Agreement between telephone and face-to-face interviews was contrasted with interrater values, which were obtained by having a second interviewer rate a recording of the original interview. RESULTS: Interrater reliability was excellent. Agreement between telephone and face-to-face assessment was excellent for anxiety disorders and very good for major depressive disorder and alcohol and substance use disorders; agreement was problematic, however, for adjustment disorder with depressed mood. Strong support was shown for the validity of the axis II telephone assessment format. Small but consistent trends were noted for lower rates of psychopathology reported in the second interview. CONCLUSIONS: This is the first study in which telephone and face-to-face assessments of axis I and II psychopathology were conducted with the same subjects assigned to conditions in a counterbalanced manner. The present findings provide qualified justification for the use of telephone interviews to collect axis I and II data. The apparent concerns do not appear sufficient to override the economic and logistic advantages of telephone interviewing. PMID- 9356571 TI - Clinical case: unknown. PMID- 9356572 TI - Images in Psychiatry. al-Razi (Rhazes), 865-925. PMID- 9356573 TI - Basilar artery response to hyperventilation in panic disorder. AB - OBJECTIVE: The purpose of this study was to measure the response of basilar artery blood flow to hyperventilation in patients with panic disorder. METHOD: Transcranial Doppler ultrasonography was used to measure basilar artery flow during rest and after hyperventilation in 16 patients with panic disorder and eight normal comparison subjects. The subjects rated their dizziness at each phase. RESULTS: The patients with panic disorder demonstrated greater reduction in flow rates and greater increases in dizziness than the normal comparison subjects. CONCLUSIONS: The greater basilar artery sensitivity to hyperventilation shown by panic disorder patients suggests a possible mechanism for the development of neurological symptoms during panic attacks. PMID- 9356574 TI - Long-term mental health effects of the Chernobyl disaster: an epidemiologic survey in two former Soviet regions. AB - OBJECTIVE: This study assessed the long-term mental health effects of the nuclear accident at Chernobyl. METHOD: Two population samples (N = 3,044), one from the Gomel region, close to the accident site, and one from Tver, 500 miles away, were studied 6 1/2 years after the event with the use of a variety of self-report questionnaires and a standardized psychiatric interview. RESULTS: The prevalence of psychological distress and DSM-III-R psychiatric disorders was exceptionally high in both regions. Scores on the self-report scales were consistently higher in the exposed region; however, a higher risk of DSM-III-R psychiatric disorders could be demonstrated only among women with children under 18 years of age in the exposed region. CONCLUSIONS: A substantial long-term mental health effect of the Chernobyl incident was demonstrated, mainly at a subclinical level. PMID- 9356575 TI - Possible discrimination in recruitment of psychiatry residents? AB - OBJECTIVE: The authors sought to determine whether there is a selection bias against international medical graduate applicants for U.S. residency training positions in psychiatry. METHOD: Identical requests for a program application were sent by two resident applicants--one international medical graduate and one graduate of a U.S. medical school--to 193 residency training programs, and the rate and character of responses were compared. RESULTS: The response rate to requests for an application form was significantly higher for the U.S. medical school graduate (159 responses) than the international medical graduate (87 responses). The quality of responses was also different in some cases. CONCLUSIONS: Some residency programs in psychiatry are attempting to limit the influx of international medical graduate applicants at the very first level: the request for an application form. The reasons for this practice are not known, but discrimination could be a possible explanation. PMID- 9356576 TI - One-year follow-up of secondary versus primary mental disorder in persons with comorbid substance use disorders. AB - OBJECTIVE: Service utilization and outcomes of dually diagnosed patients with independent mental disorders and those with substance-induced mental disorders were compared. METHOD: Diagnosis, service use, and severity of substance use problems at baseline and 1 year later were assessed in consecutively admitted inpatients with independent mental disorders plus substance use disorders (N = 71), substance-induced mental disorders plus substance use disorders (N = 38), and independent mental disorders only (N = 59). RESULTS: At follow-up, patients with substance-induced mental disorders at baseline were more likely to have been rehospitalized than the other groups, were more likely to have used outpatient substance abuse services, were less likely to have an independent mental disorder, and had the most severe alcohol- and drug-related impairment. CONCLUSIONS: Treatment programs for both types of dual diagnosis patients must address mental health concerns. PMID- 9356577 TI - Cognitive impairment in adolescents with schizophrenia. AB - OBJECTIVE: The purpose of this study was to determine whether adolescent schizophrenia is characterized by neuropsychological deficits. METHOD: The performance on a battery of neuropsychological tests of 17 adolescents with schizophrenia (mean age = 15.71 years) was compared with that of 17 normal adolescents (mean age = 15.12 years). RESULTS: Compared with the normal subjects, the patients were impaired on 10 of the 13 measures; larger effect sizes were shown for measures involving working memory and attention than for those involving secondary memory, generative naming, and executive functions. CONCLUSIONS: Adolescents with schizophrenia have generalized cognitive dysfunction, which is most apparent on tests of attention and working memory. PMID- 9356579 TI - Psychotic episode associated with dexfenfluramine. PMID- 9356578 TI - Tramadol-induced mania. PMID- 9356580 TI - Treatment of psychotic depression. PMID- 9356581 TI - High-dose olanzapine for treatment-refractory schizophrenia. PMID- 9356582 TI - Androgenic activity in autism. PMID- 9356583 TI - Compliance complications in cardiac patients. PMID- 9356584 TI - Consistency of traumatic memories. PMID- 9356585 TI - Consistency of traumatic memories. PMID- 9356586 TI - Borderline-dissociation comorbidity. PMID- 9356587 TI - Urge to splurge. PMID- 9356588 TI - Repetitive behaviors and D8/17 positivity. PMID- 9356589 TI - High sucrose preference in alcoholic men. PMID- 9356590 TI - Compulsive sexual behavior characteristics. PMID- 9356591 TI - HIV and depression. PMID- 9356593 TI - Obsessive-compulsive symptoms in schizophrenia. PMID- 9356594 TI - Religion and psychopathology. PMID- 9356592 TI - Disputing psychiatry's redefinition. PMID- 9356595 TI - Antimicrobial prophylaxis in surgery. PMID- 9356596 TI - Prevention of bacterial endocarditis. PMID- 9356597 TI - The heart and conduit vessels in hypertension. AB - The heart and conduit vessels, integral components of a pulsatile pumping system, undergo complex adaptive and degenerative changes in response to the increased load of hypertension. Over the last two decades, great technological strides have been made with regards to further discovering the role of the heart and conduit vessels in hypertension. This article reviews the adaptation of the heart and vessels to hypertension, the clinical implications of these structural and functional changes, and the effects of therapy on cardiovascular function. PMID- 9356598 TI - Nephrosclerosis and hypertension. AB - In patients with benign nephrosclerosis, the histologic changes are characterized by hyaline degeneration of afferent arterioles with reduced kidney size. Although the glomeruli are nearly intact in patients with adult essential hypertension, the greatest numbers of sclerotic glomeruli are seen in nephrosclerosis with the aging process. Aging undoubtedly plays a role. In the authors' experience, the kidney of an elderly subject, although with normotensive pressure and normal level of cholesterol, shows an increased mesangial matrix and hypertrophic vascular medial smooth muscle cells. Kidneys of elderly subjects also are associated with a large number of sclerotic glomeruli. Experimental evidence supports the notion that the pathogenesis of glomerulosclerosis with nephrosclerosis has been demonstrated as important factors: (1) the elevation of PG (glomerular hypertension); (2) mesangial dysfunction, such as mesangiolysis and increased mesangial matrix; and (3) genetic abnormalities (apoptosis) in mesangial cells with glomerular hypertension. Malignant nephrosclerosis is characterized histologically by vascular endothelial damage and fibrinoid necrosis of afferent and interlobular arteries. In an afferent arteriole or a glomerulus, NOS or endothelin produced in endothelial cells may play a role in the reduction or the maintenance of vascular tone. The frequency of malignant hypertension has decreased because of the effective treatment of essential hypertension with new antihypertensive agents: calcium antagonists, ACE inhibitors, and angiotensin II receptor antagonists. Therefore, the importance of the prevention of essential hypertension with these antihypertensive agents, by slowing and stopping the increase in blood pressure from mild hypertension, has received widespread recognition in the prevention of organ damage, such as cases of cardiovascular disease and ESRD. Thus, prevention of renal injuries is an important goal of antihypertensive therapy. PMID- 9356599 TI - Nonpharmacologic approaches to hypertension. Weight, sodium, alcohol, exercise, and tobacco considerations. AB - Weight loss decreases blood pressure, and this change can be sustained over the long-term when the lower weight is maintained. Salt restriction may be effective in blood pressure control only in salt-sensitive individuals. Heavy drinkers (those who drink more than three drinks [30 mL] daily) experience deleterious effects such as hypertension and more cardiovascular risk factors. Consequently, they should be advised to reduce alcohol intake to less than 30 mL daily. Endurance training with dynamic exercise appears to be beneficial for hypertensive patients, although recommendation guidelines are still imprecise. Finally, smoking cessation has not been proven to decrease blood pressure levels but should nonetheless be recommended because of its favorable effects on cardiovascular morbidity and mortality. PMID- 9356600 TI - Current status of diuretics, beta-blockers, alpha-blockers, and alpha-beta blockers in the treatment of hypertension. AB - The article describes the current status of four main antihypertensives. Diuretics are making a bit of a comeback after seeing their popularity wane during the 1980s. beta-blockers also saw a bit of a popularity decrease in the 1980s due to some adverse side effects which the author feels were somewhat exaggerated. alpha-blockers have yet to be particularly successful in the treatment of hypertension, due to adverse side effects. alpha-beta-blockers appear to hold significant promise in the further treatment of hypertension. PMID- 9356601 TI - Antihypertensive therapy. Angiotensin-converting enzyme inhibitors, angiotensin II receptor antagonists, and calcium antagonists. AB - ACEIs, angiotensin II receptor antagonists, and calcium antagonists are effective and well-tolerated antihypertensive agents but, except in special situations, should be considered alternative drugs for first line therapy until randomized trials show that they are at least as effective as diuretics and beta-blockers in preventing cardiovascular morbidity and mortality for a broad spectrum of hypertensive patients. ACEIs are particularly indicated for managing patients with congestive heart failure due to systolic dysfunction and patients with diabetic nephropathy, especially in Type I diabetes. Theoretically, the AII receptor antagonists will be equally effective for these indications, and randomized trials are now underway to demonstrate this. Special indications for calcium antagonists in the management of hypertension include angina pectoris, and for the non-dihydropyridine calcium antagonists, paroxysmal supraventricular tachycardia, and atrial fibrillation with rapid ventricular rate. Isolated systolic hypertension in the elderly is a special indication for long-acting dihydropyridine calcium antagonists, although diuretics are preferred. Calcium antagonists have been particularly effective in managing hypertension induced by cyclosporine. They are contraindicated in CHF due to systolic dysfunction and in the management of acute myocardial infarction. The long-term cardioprotective effect of calcium antagonists after a myocardial infarction has been demonstrated only for verapamil and diltiazem in patients with no evidence of LV dysfunction during their infarction. Calcium antagonists should be prescribed for this purpose only when beta-blockers are poorly-tolerated or contraindicated. PMID- 9356602 TI - Hypertension in special populations. AB - Essential hypertension is a common disorder, and a variety of antihypertensive drugs are available to lower blood pressure in the normal range. Identifying special subpopulations by differences in age, gender, race, and body weight has taught clinicians to be more selective in antihypertensive therapy. The rationale for this selectivity is often speculative and has not been corroborated by any hard data. It is hoped that some of the prospective, randomized trials currently in progress will throw some light on this question. PMID- 9356603 TI - Cyclosporine-induced hypertension in cardiac transplantation. AB - Cyclosporine-induced hypertension occurs in more than 90% of patients following cardiac transplantation. This article underlines the clinical characteristics as well as the mechanisms that can be associated with the development of cyclosporine-induced hypertension. In addition, the clinical trials up to date for the treatment of hypertension following cardiac transplantation are discussed. However, in view of the possible long-term sequelae associated with cyclosporine-induced hypertension, further studies to evaluate the long-term efficacy and safety of antihypertensive agents and finally the long-term effects of hypertension on the cardiac allograft are needed. PMID- 9356604 TI - New developments in drug therapy of hypertension. AB - The explosion of new knowledge about the complex mechanisms mediating high blood pressure is providing new targets for drug therapy of hypertension and other cardiovascular disorders. This article reviews the current status of several new approaches in the management of hypertension, including vasopressin antagonists, natriuretic peptide clearance inhibitors, endothelin antagonists, renin inhibitors, angiotensin receptor antagonists, and selective T-type calcium ion channel antagonists. PMID- 9356605 TI - Accountability in managed health care. PMID- 9356606 TI - CT of blunt abdominal trauma in adults. PMID- 9356607 TI - Treatment of peripheral vascular disease with stent-grafts. PMID- 9356608 TI - Diagnosis of hyperacute ischemic infarct with CT: key to improved clinical outcome after intravenous thrombolysis? PMID- 9356609 TI - Liver-specific contrast media: a magic bullet or a weapon for dedicated targets? PMID- 9356610 TI - A second look at the second-look angiogram in cases of subarachnoid hemorrhage. PMID- 9356611 TI - Acute stroke: usefulness of early CT findings before thrombolytic therapy. AB - PURPOSE: To determine whether the extent of subtle parenchymal hypoattenuation detected on computed tomographic (CT) scans obtained within 6 hours of ischemic stroke is a factor in predicting patients' response to thrombolytic treatment. MATERIALS AND METHODS: The baseline CT scans of 620 patients, who received either recombinant tissue plasminogen activator (rt-PA) or a placebo, in a double-blind, randomized multicenter trial were prospectively evaluated and assigned to one of three categories according to the extent of parenchymal hypoattenuation: none, 33% or less (small), or more than 33% (large) of the middle cerebral artery territory. The association between the extent of hypoattenuation on the baseline CT scans and the clinical outcome in the placebo-treated and the rt-PA-treated groups after 3 months was analyzed. RESULTS: In 215 patients with a small hypoattenuating area, treatment increased the chance of good outcome. In 336 patients with a normal CT scan and in 52 patients with a large hypoattenuating area, rt-PA had no beneficial effect but increased the risk for fatal brain hemorrhage. CONCLUSION: The response to rt-PA in patients with ischemic stroke can be predicted on the basis of initial CT findings of the extent of parenchymal hypoattenuation in the territory of the middle cerebral artery. PMID- 9356612 TI - Intracranial aneurysm: anatomic factors that predict the usefulness of intraoperative angiography. AB - PURPOSE: To correlate the size and location of intracranial aneurysm with the need to reposition the aneurysm clip after intraoperative angiography. MATERIALS AND METHODS: In 199 consecutive patients with 234 clipped intracranial aneurysms, 273 intraoperative angiographic studies were retrospectively reviewed. Aneurysm size and location, determined with preoperative angiographic and surgical reports, were correlated with the frequency of clip repositioning because of parent- or branch-vessel compromise or unexpected residual aneurysm. RESULTS: Findings from intraoperative angiograms resulted in clip repositioning in 46 of 273 (16.8%) studies. Clip repositioning was statistically significantly less frequent with aneurysms of the posterior communicating (three of 52 [5.7%] studies) and anterior choroidal (none of 12 studies) arteries. High rates of clip repositioning were found in aneurysms of the superior hypophyseal artery (seven of 18 [38.9%] studies), superior cerebellar artery (three of five [60.0%] studies), and bifurcation of the internal carotid artery (three of nine [33.3%] aneurysms). In 98 conventional follow-up angiographic studies, seven (7%) false negative cases with unsuspected aneurysm neck remnant were found. CONCLUSION: The rate of clip repositioning in aneurysms of the posterior communicating or anterior choroidal arteries was less than that at other locations (P < .05). Intraoperative angiography may not be necessary when aneurysms are at these two locations. PMID- 9356613 TI - Evaluation of abdominal aortic aneurysm for stent-graft placement: comparison of gadolinium-enhanced MR angiography versus helical CT angiography and digital subtraction angiography. AB - PURPOSE: To determine whether magnetic resonance (MR) angiography can be used alone to evaluate abdominal aortic aneurysms (AAAs) for endovascular placement of stent grafts. MATERIALS AND METHODS: Sixty-one patients with AAAs underwent gadolinium-enhanced MR angiography of the abdominal aorta and pelvic arteries. Measurements of the size and extent of the AAAs were compared with helical computed tomographic (CT) and digital subtraction angiographic measurements; 95% confidence intervals for the differences in the means were determined. RESULTS: Because of the larger field of view, MR angiography was superior to CT angiography in assessing visceral iliac artery disease. Both modalities were equal in evaluating the proximal extent of the AAA (mean difference, -0.16 mm; 95% CI, -0.31, 0.64) and in measuring all aortic dimensions (e.g., mean difference in the proximal neck diameter, -0.74 mm; 95% CI, -0.98, -0.49). MR angiography was inferior to CT angiography in depicting accessory renal arteries (seven of 12) and in grading renal artery stenoses (sensitivity, 100% [95% CI, 0.90, 1.00]; specificity, 84% [95% CI, 0.74, 0.91]). CONCLUSION: Gadolinium enhanced MR angiography is a fast, reliable means of providing all the information relevant to the preoperative assessment of endovascular aortic stent graft placement. PMID- 9356614 TI - US-based evaluation of hemodynamic parameters in the common carotid artery: a nomogram trial. AB - PURPOSE: To evaluate hemodynamic data of the common carotid artery (CCA) with an ultrasound (US)-based technique in an age- and sex-matched healthy population. MATERIALS AND METHODS: Measurements from a color M-mode tracing of each CCA in 205 healthy subjects (age range, 20-87 years) were performed with a time domain processing system. Lumen change of the color M-mode tracing, flow velocity, volume flow rate, and diastolic volume index (DVI) were calculated five times. Blood pressure was measured after the US examination. Fifteen subjects were excluded from the nomogram trial because of hypertension, and 12 and four subjects were excluded because of an asymptomatic carotid artery stenosis or atrial fibrillation, respectively. RESULTS: Mean volume flow rate in the CCA of the remaining 174 healthy volunteers was 415.0 mL/min +/- 87.0 (men [n = 84], 441.0 mL/min +/- 95.5; women [n = 90], 390.2 mL/min +/- 70.5). The following statistically significant associations were noted: between vessel lumen and sex and blood pressure; between peak systolic velocity and sex and age; between volume flow rate and sex, age, and systolic blood pressure; and between DVI and age, heart rate, and systolic and diastolic blood pressure. CONCLUSION: Time domain processing is an intrasessionally well-reproducible US method for measuring velocities and volume flow in the CCA. PMID- 9356615 TI - Gd-EOB-DTPA and Yb-EOB-DTPA: two prototypic contrast media for CT detection of liver lesions in dogs. AB - PURPOSE: To characterize computed tomographic (CT) attenuation of iodine, gadolinium, and ytterbium in vitro and to study CT liver enhancement after administration of two prototypic hepatocyte-directed contrast media in dogs. MATERIALS AND METHODS: Samples with increasing concentrations of iodine, gadolinium, and ytterbium were measured for CT attenuation in a water phantom at tube voltages of 80, 120, and 137 kV. Three groups of five adult beagle dogs each received a 0.5 mmol/kg dose of either gadoxetic acid disodium (gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid [EOB-DTPA]) or Yb-EOB-DTPA as intravenous infusions or Yb-EOB-DTPA as an intravenous bolus injection. RESULTS: At 120 kV, in vitro CT attenuation of gadolinium and ytterbium exceeded that of iodine by 41% and 45%, respectively, when measured at equal mass concentrations (in milligrams per milliliter of saline). In dogs, CT liver enhancement values above the precontrast liver attenuation were 32.6 HU +/- 2.3, 31.4 HU +/- 1.1, and 33.4 HU +/- 5.2 after 0.5 mmol/kg Gd-EOB-DTPA or Yb-EOB-DTPA infusion or Yb EOB-DTPA bolus, respectively. The values were not significantly different (P > .05) CONCLUSION: Both Gd-EOB-DTPA and Yb-EOB-DTPA provide excellent CT liver enhancement in dogs. CT liver enhancement is not significantly altered by either the absorbing element (gadolinium vs ytterbium) or the mode of application (infusion vs bolus). PMID- 9356616 TI - Hepatic metastases: percutaneous radio-frequency ablation with cooled-tip electrodes. AB - PURPOSE: To assess the feasibility and safety of using cooled-tip electrodes to increase the volume of coagulation necrosis obtained or reduce the number of treatment sessions necessary with percutaneous tumor radio-frequency (RF) ablation. MATERIALS AND METHODS: Twenty-nine patients with 44 hepatic metastases (1.3-5.1 cm diameter) from colorectal (n = 22), gastric (n = 5), pancreatic (n = 1), or breast (n = 1) carcinoma were treated with RF ablation using cooled-tip, 18-gauge electrodes with 2-3 cm tip exposure. Each tumor was treated in one or two treatment sessions. RESULTS: Technical success, ablation of all visualized tumor, was achieved in 40 (91%) metastases. Findings at computed tomography (CT) and magnetic resonance (MR) imaging performed 3-6 months after treatment confirmed complete necrosis of the entire metastasis in 66%. Disease-free survival was 50% at 12 months and 33% at 18 months, with localized progression of disease in 34% of treated lesions. Overall survival was 100%, 94%, and 89% at 6, 12, and 18 months, respectively. Only one complication, self-limited hemorrhage, was seen. CONCLUSION: Use of cooled-tip electrodes was a safe and feasible adjunct for tumor RF ablation therapy that produced larger volumes of coagulation necrosis with fewer electrode insertions than is produced with other RF ablation techniques. PMID- 9356617 TI - Femoropopliteal artery stent placement: evaluation of long-term success. AB - PURPOSE: To evaluate the long-term success of femoropopliteal artery stent placement after technical failure of balloon angioplasty and to determine potential factors predictive of stent failure. MATERIALS AND METHODS: In 80 patients, flexible tantalum stents were implanted for treatment of residual stenosis greater than 30% after balloon angioplasty of femoropopliteal artery stenoses (n = 33) or occlusions (n = 47). Follow-up examinations were performed at 1-72 months to evaluate early and late stent failure according to risk factors such as lesion type, anatomic location, stent length, runoff quality, and restenosis after balloon angioplasty. RESULTS: Early stent thrombosis occurred within 10 days after stent placement in four (5%) patients; the frequency was higher (P < .01) in stents longer than 4 cm (14%) versus that in stents 4 cm or shorter (2%). For all stents, the 2-year primary patency rate was 51% and the 3 year rate was 48%. The 2-year primary patency rate was significantly better (P < .05) for shorter stents (59%) versus that for longer stents (30%) and for treated stenoses (73%) versus treated occlusions (33%). Multivariate analysis revealed that lesion type and lesion length were the most reliable factors predictive of stent failure. CONCLUSION: Successful stent therapy does not seem to depend on the type of stent used but rather on lesion type, lesion length, and other predisposing factors. PMID- 9356618 TI - Patient dose in radiologically guided interventional vascular procedures: conventional versus digital systems. AB - PURPOSE: To calculate the difference in the patient radiation dose in radiologically guided interventional vascular procedures between conventional and digital systems and to estimate the effective dose and the energy imparted with the digital system. MATERIALS AND METHODS: A total of 318 procedures (in 318 patients) in 15 different examination groups were analyzed. The dose-area product was determined by using a transmission chamber fitted to an x-ray-tube light-beam diaphragm; the effective dose was determined by using software. RESULTS: Urinary and biliary tract procedures showed small differences in the average dose-area product between conventional and digital systems. The dose-area products in the vascular procedures were higher with the digital than with the conventional system. The average effective dose and energy imparted were 0.88 mSv and 129 mJ, respectively, in the subcutaneous placement of a reservoir for analgesic administration and as much as 25.7 mSv and 829 mJ, respectively, in spermatic vein embolization. CONCLUSION: The dose-area product was higher with the digital system than with the conventional system in 13 of the 15 groups. To reduce the patient dose in vascular interventional radiology procedures, the training of personnel and the frequent use of conventional fluoroscopy and low-dose imaging are required. PMID- 9356619 TI - Entrance skin exposure and mean glandular dose: effect of scatter and field gradient at mammography. AB - PURPOSE: To investigate the effect of scatter and x-ray field gradient on entrance skin exposure and mean glandular dose by using the measurement protocol of the American College of Radiology. MATERIALS AND METHODS: The exposure difference between the off-axis geometry of the American College of Radiology protocol and the central-axis geometry was measured by using eight common ionization chambers and two mammographic units (Senographe DMR; GE Medical Systems, Milwaukee, Wis; and Mam-CP II; Transcontinental X-ray, Charlotte, NC) at 25, 28, and 30 kVp. The field gradient was also measured on film with a scanning densitometer. RESULTS: The central-axis exposure was 2.0%-3.6% higher than the off-axis exposure for the Senographe DMR unit and 3.1%-8.0% higher for the Mam-CP II unit. The maximum phantom scatter was 0.7% (Senographe DMR) and 1.3% (Mam-CP II). CONCLUSION: Entrance skin exposure varies, depending on the type of ionization chamber and the characteristics of the mammographic unit. Determination of mean glandular dose with the American College of Radiology protocol may lead to underestimates of the actual measured dose by 4%-8% for the x-ray machines used in this study. PMID- 9356622 TI - Clip placement after stereotactic vacuum-assisted breast biopsy. AB - PURPOSE: To assess accuracy and usefulness of placement of a localizing clip after stereotactic, vacuum-assisted breast biopsy. MATERIALS AND METHODS: Retrospective review was performed of 57 lesions that underwent placement of a localizing clip after stereotactic vacuum-assisted biopsy with an 11-gauge (n = 42) or 14-gauge (n = 15) probe. The clip was placed when images obtained after stereotactic biopsy suggested that the lesion seen at mammography was removed. Coordinates of the clip on stereotactic images obtained after placement were compared with lesion coordinates determined before biopsy. Surgery was performed in 25 cases. Mammographic and histopathologic findings were reviewed. RESULTS: The distance from clip to lesion site was less than 1 cm in 40 (95%) of 42 lesions that underwent clip placement with the 11-gauge probe versus 11 (73%) of 15 lesions that underwent clip placement after 14-gauge biopsy (P < .04). The biopsy site was identified in the surgical specimen in 19 (100%) lesions with clips after 11-gauge biopsy and five (83%) of six lesions with clips after 14 gauge biopsy. No complications occurred. CONCLUSION: A localizing clip can be placed in proximity to the stereotactic biopsy site through an 11-gauge probe. Clip placement can enable accurate localization for surgical excision. PMID- 9356623 TI - Canceled stereotactic core-needle biopsy of the breast: analysis of 89 cases. AB - PURPOSE: To determine reasons for cancellation of stereotactic core-needle breast biopsy and outcome in canceled cases. MATERIALS AND METHODS: Among 572 scheduled stereotactic core-needle biopsies, 89 cases (16%) in 88 patients were canceled. In canceled cases, mammogram origin, mammographic abnormality, reason for cancellation, and outcome were determined. RESULTS: In canceled cases, 50 (57%) of 88 patients were referred from another facility. Mammographic abnormality in most cases (72 [81%] of 89 canceled biopsies) was a mass(es); calcifications occurred in 14 cases (16%). Reasons for cancellation included (a) lesion was not recognized (26 cases [29%]), (b) lesion was reassessed as benign (17 cases [19%]), (c) cysts were diagnosed with ultrasound (US) (12 cases [13%]) or aspiration (11 cases [12%]), (d) lesion location was suboptimal (12 cases [13%]), (e) patient was intolerant of procedure (seven cases [8%]), and (f) other (four cases [4%]). Numbers of canceled biopsies from another facility and those from the authors' institution differed in cases in which lesions were reassessed as benign (12 and five cases, respectively) or cysts were diagnosed with US (10 and two cases, respectively). Lesions that could not be targeted included many pseudolesions and three cancers. CONCLUSION: Complete work-up, including US examination, of breast lesions is necessary before stereotactic core-needle biopsy is scheduled. Inability to recognize a suspected lesion on stereotactic images should not preclude biopsy with another method. PMID- 9356621 TI - Tissue marking clip for stereotactic breast biopsy: initial placement accuracy, long-term stability, and usefulness as a guide for wire localization. AB - PURPOSE: To determine initial placement accuracy, long-term stability, and usefulness as a guide for wire localization for metallic marker clips placed percutaneously after stereotactic breast biopsy. MATERIALS AND METHODS: One hundred forty-nine marker clips were placed percutaneously with a straight-needle or through-probe method, and clip positions were measured. The locations of 31 marker clips were followed up from deployment to first follow-up mammography. Thirty-six biopsy sites with marker clips were excised surgically and examined; 18 of these marker clips were targets for wire localization. The locations of 22 benign lesions were measured over time to calibrate the measurement system. RESULTS: Baseline variability was 8 mm. Initial marker clip deployment averaged 5 mm above baseline from the center of the target lesion (P < or = .01). Compared with baseline variability, marker clips remained in place from initial deployment to first imaging follow-up (mean, 8.6 months). Potentially clinically meaningful misplacement rates (deployment > 24 mm from target lesion center) were 7% for the through-probe method and 11% for the straight-needle method (not significantly different; P = .33). CONCLUSION: The marker clips appear to be useful targets for wire localization when the entire target lesion is removed at directional, vacuum assisted breast biopsy. Upright, two-view mammography is recommended after deployment of the marker clip to document location. PMID- 9356620 TI - Digital tomosynthesis in breast imaging. AB - PURPOSE: To describe and evaluate a method of tomosynthesis breast imaging with a full-field digital mammographic system. MATERIALS AND METHODS: In this tomosynthesis method, low-radiation-dose images were acquired as the x-ray source was moved in an arc above the stationary breast and digital detector. A step-and expose method of imaging was used. Breast tomosynthesis and conventional images of two imaging phantoms and four mastectomy specimens were obtained. Three experienced readers scored the relative lesion visibility, lesion margin visibility, and confidence in the classification of six lesions. RESULTS: Tomosynthesis image-reconstruction algorithms allow tomographic imaging of the entire breast from a single arc of the x-ray source and at a radiation dose comparable with that in single-view mammography. Except for images of a large mass in a fatty breast, the tomosynthesis images were superior to the conventional images. CONCLUSION: Digital mammographic systems make breast tomosynthesis possible. Tomosynthesis may improve the specificity of mammography with improved lesion margin visibility and may improve early breast cancer detection, especially in women with radiographically dense breasts. PMID- 9356624 TI - Breast carcinoma: MR imaging before re-excisional biopsy. AB - PURPOSE: To investigate the role of magnetic resonance (MR) imaging in the examination of patients after excisional biopsy of breast carcinoma before re excision. MATERIALS AND METHODS: Forty-seven patients underwent contrast material enhanced MR imaging after initial excisional biopsy of breast carcinoma before further surgery. RESULTS: The positive predictive value of MR imaging for predicting residual disease was 82%; the negative predictive value was 61%. Fourteen patients had multifocal (n = 6) or diffuse (n = 8) carcinoma. The extent of tumor was correctly identified with MR imaging alone in nine of the 14 patients, with both mammography and MR imaging in three patients, with mammography alone in one patient, and with no imaging modality in one patient. In four of the 14 patients, management was altered from re-excision to mastectomy (n = 3) or from breast-conservation therapy to mastectomy (n = 1). CONCLUSION: MR imaging has a high positive predictive value for predicting residual tumor after excisional biopsy. The identification of mammographically and clinically unsuspected multifocal or extensive residual tumor may lend support for mastectomy rather than re-excision. However, false-negative findings due to postsurgical changes and false-positive findings due to enhancement of granulation tissue and benign breast tissue remain limitations. PMID- 9356625 TI - MR imaging of the breast in patients with occult primary breast carcinoma. AB - PURPOSE: To evaluate the utility of contrast material-enhanced magnetic resonance (MR) imaging of the breast for localization of the site of primary breast carcinoma in patients with isolated ipsilateral axillary metastasis, without other focal findings at physical examination or mammography. MATERIALS AND METHODS: Twelve women with biopsy-proved metastatic adenocarcinoma to axillary lymph nodes and occult primary tumor underwent MR imaging at 1.5 T with a three dimensional spoiled gradient-echo pulse sequence before and after administration of gadopentetate dimeglumine. Enhancing areas were correlated with histopathologic findings at mastectomy (n = 8) or breast-conserving treatment (n = 4). RESULTS: In nine (75%) of the 12 patients, enhancement was seen on MR images that represented the site of the primary tumor at surgery. Eight had focal masses, and one had an area of regional enhancement. Two patients had negative MR findings, and no tumor was found at mastectomy. One patient with regional enhancement on MR images had no corresponding tumor at surgery. CONCLUSION: Breast MR imaging enables identification of the site of primary tumor in most patients suspected of having occult primary breast cancer, and MR findings can influence surgical treatment. PMID- 9356626 TI - Screening mammography: experience in a health maintenance organization. AB - PURPOSE: To evaluate (a) the relationship between mammogram interpretation and diagnosis of new breast cancer and (b) interprovider variation in mammogram interpretation. MATERIALS AND METHODS: Interpretations of screening mammograms (133,668 mammograms in 114,899 women) acquired during 21 months in a large health maintenance organization were categorized (categories 1-5) with use of a standard format. During 1 year after mammography, new breast cancer was identified with use of claims data. Interprovider variation in the categories read was evaluated, and percentages of these categories were correlated with breast cancer detection. RESULTS: Over the 21 months, 1,018 mammograms were followed by a diagnosis of new breast cancer. The category of mammogram interpretation was strongly associated with the diagnosis of new breast cancer; in 47.5% cases of category 5 mammograms, breast cancer was diagnosed. There was substantial interprovider variation in the percentages of category 3, 4, or 5 mammograms read. The percentage of category 4 and 5 mammograms read correlated inversely with the likelihood of cancer detection (Pearson correlation coefficient [r] = -.4778 after log-log transformation, P < .001). CONCLUSION: A strong correlation existed between a mammographic abnormality suggestive of cancer and its detection; however, substantial interprovider variation in the reading of category 3, 4, and 5 mammograms and their positive predictive values existed. Reduction of interprovider variation should improve quality of care because the number of false-negative and false-positive mammograms should decrease. PMID- 9356628 TI - Acute pulmonary embolism: role of helical CT in 164 patients with intermediate probability at ventilation-perfusion scintigraphy and normal results at duplex US of the legs. AB - PURPOSE: To assess prospectively the clinical effectiveness of helical computed tomography (CT) in the evaluation of patients with unresolved suspicion for pulmonary embolism (PE). MATERIALS AND METHODS: Helical CT was performed in 164 consecutive patients suspected of having acute PE, intermediate probability at ventilation-perfusion (V-P) scintigraphy, and normal findings at duplex ultrasonography (US) of the legs. Fifteen patients also underwent pulmonary angiography. Helical CT results were analyzed immediately to help plan anticoagulant treatment. If helical CT did not show PE, anticoagulant treatment was not indicated. Clinical outcome for these patients was assessed during 3 month follow-up. RESULTS: In 40 (24.4%) of 164 patients, the diagnosis of PE was based on results at helical CT (n = 39) or pulmonary angiography (n = 1). Repeated Doppler US of the legs depicted one thrombus in the calf of three patients with normal results at helical CT that could have been responsible for PE. During 3-month follow-up, three patients experienced recurrent PE (one death, two recurrences). Therefore, PE occurred in six (5.4% [95% confidence interval, 1.3%, 9.7%]) of 112 patients with normal findings at helical CT who did not receive anticoagulant treatment. CONCLUSION: Findings at helical CT allowed accurate diagnosis of acute PE in patients with intermediate probability at V-P scintigraphy and without deep venous thrombosis at duplex sonography of the legs. PMID- 9356627 TI - Pulmonary embolism: prospective comparison of spiral CT with ventilation perfusion scintigraphy. AB - PURPOSE: To compare prospectively the accuracy of spiral computed tomography (CT) with that of ventilation-perfusion scintigraphy for diagnosing pulmonary embolism. MATERIALS AND METHODS: Within 48 hours of presentation, 142 patients suspected of having pulmonary embolism underwent spiral CT, scintigraphy, and (when indicated) pulmonary angiography. Pulmonary angiography was attempted if interpretations of spiral CT scans and of scintigrams were discordant or indeterminate and intermediate-probability, respectively. RESULTS: In the 139 patients who completed the study, interpretations of spiral CT scans and of scintigrams were concordant in 103 patients (29 with embolism, 74 without). In 20 patients, intermediate-probability scintigrams were interpreted (six with embolism at angiography, 14 without); diagnosis with spiral CT was correct in 16. Interpretations of spiral CT scans and those of scintigrams were discordant in 12 cases; diagnosis with spiral CT was correct in 11 cases and that with scintigraphy was correct in one. Spiral CT and scintigraphic scans of four patients with embolism did not show embolism. Sensitivities, specificities, and kappa values with spiral CT and scintigraphy were 87%, 95%, and 0.85 and 65%, 94%, and 0.61, respectively. CONCLUSION: In cases of pulmonary embolism, sensitivity of spiral CT is greater than that of scintigraphy. Interobserver agreement is better with spiral CT. PMID- 9356629 TI - HIV-associated human herpesvirus 8-positive primary lymphomatous effusions: radiologic findings in six patients. AB - PURPOSE: To define the imaging features of body cavity-based lymphoma (BCBL) related to the acquired immunodeficiency syndrome. This is a peculiar type of non Hodgkin lymphoma harboring infection by human herpesvirus 8 (HHV-8) and displaying a peculiar tropism for the serous body cavities. MATERIALS AND METHODS: At diagnosis of BCBL, six consecutive patients were investigated with radiography of the chest and conventional computed tomography (CT) of the chest and abdomen. For all patients, clinical features and results of biologic characterization of the lymphoma were also available. RESULTS: In all patients, chest radiographs displayed bilateral or unilateral pleural effusion in the absence of parenchymal opacities or mediastinal enlargement. CT scans confirmed chest radiographic findings and revealed a slight thickening of the parietal pleura in all patients and a pericardial thickening in four patients. CT also depicted pericardial and abdominal effusions in five and two patients, respectively, in the absence of solid tumor masses or parenchymal abnormalities. CONCLUSION: BCBL must be included among the differential diagnoses of serous effusions detected radiologically in individuals infected with the human immunodeficiency virus (HIV). Since the combination of serous effusion, slight serosal thickening, and absence of solid masses is compatible with, though not specific for, BCBL in the context of HIV infection, radiologic findings need to be complemented by a detailed biologic and virologic characterization of tumor cells. PMID- 9356630 TI - Mosaic attenuation pattern on thin-section CT scans of the lung: differentiation among infiltrative lung, airway, and vascular diseases as a cause. AB - PURPOSE: To determine whether infiltrative lung, airway, or vascular disease can be differentiated as the cause of mosaic attenuation on thin-section computed tomographic (CT) scans of the lung. MATERIALS AND METHODS: Thin-section CT scans were reviewed in 70 patients examined at three institutions. A mosaic attenuation pattern and pathologic or clinical proof of a specific type of disease were demonstrated. Causes of the mosaic pattern included infiltrative lung disease (n = 37), airway disease (n = 22), and vascular disease (n = 11). Thin-section CT findings were assessed independently by two observers blinded to clinical findings. RESULTS: The type of disease was identified correctly at CT in 58 (83%) of 70 patients by observer 1 and 57 (81%) of 70 patients by observer 2. Infiltrative lung disease was diagnosed correctly by both observers in 34 (92%) of 37 cases. Observer 1 identified 21 (95%) of 22 cases of airway disease and three (27%) of 11 cases of vascular disease. Observer 2 identified 19 (86%) of 22 cases of airway disease and four (36%) of 11 cases of vascular disease. CONCLUSION: Infiltrative lung disease and airway disease may be differentiated reliably as the cause of mosaic attenuation on lung CT scans, whereas vascular disease is often misinterpreted as infiltrative lung disease or airway disease. PMID- 9356631 TI - Solitary pulmonary nodules: evaluation of blood flow patterns with dynamic CT. AB - PURPOSE: To evaluate the efficacy of dynamic computed tomography (CT) for differentiating benign from malignant solitary pulmonary nodules (SPNs). MATERIALS AND METHODS: Sixty-five patients with noncalcified SPNs (diameter, < or = 30 mm; 42 malignant, 16 benign, seven inflammatory) underwent single-location dynamic contrast material-enhanced (100 mL, 4 mL/sec) serial CT. Peak height of time-attenuation curves and ratio of peak height of the SPN to that of the aorta were measured. Precontrast attenuation and enhancement pattern were recorded. Perfusion was calculated from the maximum gradient of the time-attenuation curve and the peak height of the aorta. RESULTS: Peak heights of malignant (41.9 HU +/- 2.8) and inflammatory (43.6 HU +/- 7.7) SPNs were significantly higher than that (13.4 HU +/- 2.2) of benign SPNs (P < .001; P < .01). SPN-to-aorta ratios in malignant and inflammatory SPNs were significantly higher than that in benign SPNs (P < .001, P < .05). No statistically significant differences in the peak height and SPN-to-aorta ratio were found between malignant and inflammatory SPNs. Precontrast attenuation of inflammatory SPNs was lower than that of malignant SPNs (P < .05). Perfusion values in malignant and inflammatory SPNs were significantly higher than that of the benign SPNs (P < .01). CONCLUSION: Dynamic CT provides quantitative information about blood flow patterns of SPNs and is an applicable diagnostic method for differentiating SPNs. PMID- 9356632 TI - Hiring by radiology groups in 1996. AB - PURPOSE: To determine the hiring activity of radiology groups in 1996. MATERIALS AND METHODS: A survey was mailed to a stratified, random sample of 794 radiology groups in autumn 1996. The response rate was 78%. Responses were weighted to be representative of all of the approximately 3,300 groups in the United States. Findings were compared with those of previous, similar surveys. RESULTS: In the 12 months before the survey, groups sought to hire 1,732 +/- 155 (+/- 1 standard error) diagnostic radiologists and radiation oncologists; 788 +/- 105 of these positions were to fill expansion positions, 562 +/- 86 were replacements for persons who had left the profession, and 382 +/- 78 were replacements for persons who moved to other radiology positions. During the year, an additional 500 +/- 119 positions were vacated that groups did not seek to refill. Groups succeeded in hiring 1,438 +/- 143 radiologists. The percentage of available positions that were filled did not differ across fields. Managed health care reduced the probability a group was expanding but did not otherwise affect hiring activity. CONCLUSION: The 1991-1995 decline in hiring has ceased and perhaps even reversed. There were approximately as many positions available in 1996 as were needed by graduates, although there may have been a small shortfall. PMID- 9356633 TI - Ureteropelvic junction injuries secondary to blunt abdominal trauma. AB - PURPOSE: To describe the clinical and imaging findings of ureteropelvic junction (UPJ) injuries caused by blunt trauma. MATERIALS AND METHODS: In two children (aged 10 and 16 years) and eight adults (aged 23-82 years) with UPJ injuries, findings at computed tomography (CT) (n = 10), excretory urography (n = 6), and retrograde pyelography (n = 8) were retrospectively reviewed to identify the location and extent of contrast material extravasation. Clinical and follow-up data were correlated with radiologic findings. RESULTS: CT and urography played complementary roles in diagnosis. UPJ avulsion, defined as complete transection of the ureter with no filling of the ipsilateral ureter below the level of the UPJ, was diagnosed in four patients. UPJ laceration, defined as contrast material extravasation from the UPJ with contrast material in the ipsilateral ureter distal to the point of injury, was diagnosed in six patients. Medial perirenal contrast extravasation was seen in all 10 patients but failed to help differentiate UPJ avulsion from laceration. A distinctive pattern of contrast material extravasation at CT termed "circumrenal urinoma" was present in five patients and was found to be specific for UPJ injury. CONCLUSION: Medial perinephric contrast material extravasation was highly suggestive of UPJ injury. Demonstration of ureteral filling differentiated UPJ laceration from avulsion. PMID- 9356634 TI - Normal placenta: gadolinium-enhanced dynamic MR imaging. AB - PURPOSE: To establish the appearance of the normal placenta on dynamic, gadolinium-enhanced magnetic resonance (MR) images. MATERIALS AND METHODS: Eleven patients underwent MR imaging for suspected uterine (four patients) or placental (seven patients) abnormalities (but not placental insufficiency or intrauterine growth retardation). Unenhanced or gadolinium-enhanced, T1-weighted, fat suppressed, spoiled gradient-recalled-echo MR images and T2-weighted, half Fourier, single-shot, spin-echo-train MR images were obtained. Two investigators retrospectively evaluated the images to determine the rate, pattern, and degree of placental contrast material enhancement with myometrial enhancement as a reference. RESULTS: The diagnoses were normal placenta (six patients), abnormal placental position (four patients), and subchorionic hematoma (one patient). The placenta rapidly and intensely enhanced on images acquired immediately after contrast material administration and preceded substantial enhancement of the myometrium. Immediately postcontrast images showed closely packed 2-3-cm lobules of placental enhancement in third-trimester pregnancies and heterogeneous placental enhancement in second-trimester pregnancies. In all cases, placental enhancement became more homogeneous over time. The placenta could be readily distinguished from the myometrium. All neonates were healthy with no evidence of intrauterine growth retardation. CONCLUSION: Normal placental enhancement is intense on immediately postcontrast images and precedes substantial myometrial enhancement. Third-trimester placentas exhibit a lobular pattern of enhancement, while second-trimester placentas exhibit heterogeneous enhancement. PMID- 9356635 TI - Angiomyolipoma: imaging findings in lesions with minimal fat. AB - PURPOSE: To investigate a method of diagnosing angiomyolipoma that contains minimal fat. MATERIALS AND METHODS: In six cases of angiomyolipoma with minimal fat, the attenuation on contrast material-enhanced and unenhanced computed tomographic (CT) images, the echogenicity on sonograms, the signal intensity on T2-weighted magnetic resonance (MR) images, and the gross configuration of the lesion were retrospectively analyzed. In 100 cases of renal cell carcinoma, the same parameters were analyzed, and results were compared with those of angiomyolipoma. RESULTS: When compared with the surrounding renal parenchyma, all six angiomyolipomas showed homogeneously high attenuation on unenhanced CT images, homogeneous enhancement on contrast-enhanced CT images, and homogeneous isoechogenicity on sonograms. Of the five angiomyolipomas examined with MR imaging, four were hypointense and one was isointense on T2-weighted images. All six angiomyolipomas protruded from the renal margin. None of the 100 renal cell carcinomas showed homogeneously high attenuation on unenhanced CT images, homogeneous enhancement on contrast-enhanced CT images, or homogeneous isoechogenicity on sonograms. CONCLUSION: In the kidney, homogeneously high attenuation on unenhanced CT images, homogeneous enhancement on contrast-enhanced CT images, and homogeneous isoechogenicity on sonograms are suggestive of angiomyolipoma that contains abundant muscle and minimal fat. PMID- 9356636 TI - Acute colonic diverticulitis: prospective comparative evaluation with US and CT. AB - PURPOSE: To compare the accuracy of ultrasonographic (US) and computed tomographic (CT) findings for diagnosis of acute colonic diverticulitis. MATERIALS AND METHODS: US and CT were prospectively performed in 64 consecutive patients suspected of having acute colonic diverticulitis. Images were interpreted independently in a blinded fashion. Imaging data were compared with the final diagnosis, which was based on initial clinical and follow-up examination results (n = 64) and pathologic (n = 22), endoscopic (n = 21), and contrast enema (n = 15) examination findings. RESULTS: Final diagnosis was acute colonic diverticulitis (n = 33), other acute abdominal condition (n = 24), or unknown (n = 7). Both CT and US findings yielded 84% accuracy. US and CT findings were not statistically significant different in terms of sensitivity (85% and 91%, respectively) and specificity (84% and 77%, respectively). Positive predictive value was 85% for US and 81% for CT; negative predictive value was 84% for US and 88% for CT. When determining alternative diagnoses, US and CT findings yielded sensitivity of 33% and 50%, respectively (difference not statistically significant). CT scans depicted a small pneumoperitoneum overlooked on plain radiographs and US scans. Six pericolic abscesses were depicted with both techniques; three were depicted with CT only. CONCLUSION: US and CT findings result in similar accuracy for the evaluation of patients suspected of having diverticulitis. PMID- 9356637 TI - Pancreatic malignancy: effect of dual-phase helical CT in tumor detection and vascular opacification. AB - PURPOSE: To determine the relative value of hepatic arterial and portal venous phase helical computed tomographic (CT) scans for tumor detection and vascular opacification in patients with pancreatic malignancy. MATERIALS AND METHODS: Ninety-five patients who had or were suspected of having pancreatic disease underwent dual-phase helical CT. Arterial phase scans were acquired 20-40 seconds after contrast material administration; venous phase scans, 70-100 seconds after administration. Three readers independently scored images in a blinded fashion for the presence of tumor, for lesion attenuation relative to normal pancreas, and for vascular opacification. RESULTS: The final diagnosis was pancreatic malignancy (n = 60), acute or chronic pancreatitis (n = 22), and normal pancreas (n = 13). The readers identified possible or definite tumors on arterial phase studies in 47-50 patients and on venous phase studies in 48-53 patients (P > .10). There was no statistically significant difference in tumor attenuation between scans from the two phases (P > .05). Agreement between the readers for tumor detection was not affected by the scanning phase (P > .10). Opacification of arteries and of veins was greater on arterial phase scans and on venous phase scans, respectively (P < .001). CONCLUSION: The acquisition of arterial phase scans in addition to venous phase scans does not result in improved detection of pancreatic malignancies. PMID- 9356638 TI - Intestinal ischemia in patients in whom small bowel obstruction is suspected: evaluation of accuracy, limitations, and clinical implications of CT in diagnosis. AB - PURPOSE: To determine the accuracy of computed tomography (CT) in diagnosis of intestinal ischemia in patients with possible intestinal obstruction and the limitations and clinical implications of use of CT. MATERIALS AND METHODS: In 100 patients in whom intestinal obstruction was suspected clinically, CT findings were correlated with surgical findings in 77 patients and with follow-up clinical findings after nasogastric suction in 23 patients. The interval between CT and surgical exploration in patients with ischemic bowel was 1-98 hours (mean, 13 hours). RESULTS: Correlation of CT findings of strangulation obstruction with surgical findings revealed 72 true-negative, 19 true-positive, five false positive, and four false-negative CT results. Sensitivity was 83%, specificity was 93%, accuracy was 91%, positive predictive value was 79%, and negative predictive value was 95%. CONCLUSION: CT enables accurate detection of bowel ischemia, particularly when small bowel obstruction is present. Exploratory laparotomy should be performed when unexplained disparities exist between equivocal CT findings and a deteriorating clinical condition in patients with possible small bowel obstruction or mesenteric infarction. PMID- 9356639 TI - Choledocholithiasis and bile duct stenosis: diagnostic accuracy of MR cholangiopancreatography. AB - PURPOSE: To evaluate the accuracy of magnetic resonance (MR) cholangiopancreatography for detecting bile duct calculi and stenosis. MATERIALS AND METHODS: At MR cholangiopancreatography, 108 patients suspected of having bile duct calculi or stenosis were examined with two-dimensional fast spin-echo MR sequences and respiratory gating. On the basis of findings at surgery and/or intraoperative, endoscopic retrograde, and/or percutaneous cholangiography, final diagnoses were normal bile ducts (n = 38), choledocholithiasis (n = 23), Mirizzi syndrome (n = 3), benign or malignant bile duct stenosis (n = 40), choledochal cyst (n = 1), and bile duct dilatation without calculi or stenosis (n = 3). MR cholangiopancreatographic images were analyzed retrospectively by three reviewers who were unaware of final diagnoses. RESULTS: Choledocholithiasis was diagnosed with a sensitivity of 88%-92% and a specificity of 91%-98%. False-negative readings occurred because small or impacted calculi at the distal common bile duct or ampulla were difficult to detect or distinguish from stenosis. Bile duct stenosis was diagnosed with a sensitivity of 93%-100% and a specificity of 98%. Presence or absence of bile duct abnormality was assessed with a sensitivity of 97%-99% and a specificity of 95%-97%. Interobserver agreement was very good (kappa = 0.86-0.96). CONCLUSION: With MR cholangiopancreatography, bile duct calculi and stenoses can be diagnosed with high accuracy and good interobserver agreement. PMID- 9356640 TI - Aplasia or hypoplasia of the pancreatic uncinate process: comparison in patients with and patients without intestinal nonrotation. AB - PURPOSE: To investigate whether the pancreatic uncinate process is hypoplastic in patients with intestinal nonrotation, and to evaluate the association of hypoplasia or aplasia of the uncinate process with mesenteric vascular inversion. MATERIALS AND METHODS: Computed tomographic (CT) scans of the pancreas in five patients with intestinal nonrotation and in 101 patients with normal intestinal rotation were reviewed to assess the morphology of the uncinate process. The transverse diameter of the uncinate process was measured, and the relationship with the superior mesenteric artery and vein was defined. RESULTS: The uncinate process was absent in three and unusually small in two of the five patients with nonrotation. The uncinate process was clearly present in all patients with normal rotation. The mean transverse diameter (2.60 mm +/- 2.61 [standard deviation]) of the uncinate process in patients with nonrotation was significantly smaller than that (15.1 mm +/- 5.0) in the patients with normal rotation (P < .001). Four of the five patients with nonrotation had mesenteric vascular inversion, and three of these patients had no uncinate process. None of the patients with normal rotation had mesenteric vascular inversion. CONCLUSION: The uncinate process was aplastic or hypoplastic in patients with nonrotation, which may have been associated with incomplete rotation of the ventral bud of the pancreatic primordium, as well as mesenteric vascular inversion. PMID- 9356641 TI - Primary non-Hodgkin lymphoma of the large bowel. AB - PURPOSE: To characterize the natural history of primary non-Hodgkin lymphoma of the large bowel and identify prognostic factors. MATERIALS AND METHODS: Twenty three patients with primary non-Hodgkin lymphoma according to strict criteria were identified. Seventeen patients underwent resection, and six patients underwent biopsy. Among 19 patients with intermediate- or high-grade lymphoma, 13 had diffuse large cell lymphoma. Ann Arbor stage was I in 15 cases, II in seven cases, and IV in one case. In 15 patients, the International Prognostic Index was available: 0, eight patients; 1, six patients; and 3, one patient. Postoperatively, six patients received combined chemotherapy and radiation therapy, eight patients received chemotherapy, and six patients received radiation therapy. Overall and relapse-free survival were calculated actuarially, and univariate analysis was performed with regard to stage, treatment, extent of surgery, and the International Prognostic Index. RESULTS: Median follow-up was 144 months. Two patients' disease recurred. Overall and relapse-free survival at 10 years were 61% and 82%, respectively. The International Prognostic Index was the only significant prognostic factor for overall survival (P = .03, log-rank test). CONCLUSION: The prognosis of primary non-Hodgkin lymphoma appears to be as good as that of low- or intermediate-grade lymphoma. The only significant prognostic factor for overall survival is the International Prognostic Index. PMID- 9356642 TI - The shoulder: adaptive motion correction of MR images. AB - PURPOSE: To evaluate an adaptive-motion-correction technique to reduce global motion in shoulder magnetic resonance (MR) images. MATERIALS AND METHODS: In the adaptive-motion-correction technique, interleaved navigator echoes are used to provide a measure of view-to-view displacement along the craniocaudal direction for each image echo in the acquisition. The information is then retrospectively applied to the k-space data to correct for global shoulder motion. This algorithm was evaluated in a series of 143 consecutive patient shoulder examinations by comparing the original image set for each patient with the same image set after retrospective correction by means of this algorithm. RESULTS: The average amplitude of craniocaudal motion was 1.4 mm. Image degradation due to motion was apparent in 100 (70%) of the 143 examinations. Application of the adaptive-motion correction technique improved image quality in 73 (73%) of these 100 examinations or 51% of all 143 examinations. CONCLUSION: Adaptive motion correction improved image quality in approximately three-quarters of the examinations in which motion was present. PMID- 9356643 TI - Spatial variation of T2 in human articular cartilage. AB - PURPOSE: To determine the spatial variation of in vivo cartilage T2 in young asymptomatic adults. MATERIALS AND METHODS: Quantitative T2 maps of seven asymptomatic young male adults and one male volunteer with a history of previous intraarticular chondroid fragments were calculated by using a multiecho, spin echo magnetic resonance imaging sequence at 3.0 T. The T2 maps were bilinearly interpolated to generate T2 profiles across the thickness of cartilage. RESULTS: All seven asymptomatic volunteers demonstrated a monotonic increase in T2, which increased from 32 msec +/- 1 in the deep radial zone and 48 msec +/- 1 in the deep transitional zone to 67 msec +/- 2 in the outer transitional superficial zone. The T2 profile of the volunteer with superficial fibrillation observed at arthroscopy demonstrated marked spatial heterogeneity and a statistically significant increase in cartilage T2. CONCLUSION: There is a reproducible pattern of increasing T2 that is proportional to the known spatial variation in cartilage water and is inversely proportional to the distribution of proteoglycans. The authors postulate that these regional T2 differences are secondary to the restricted mobility of cartilage water within an anisotropic solid matrix. PMID- 9356645 TI - Sciatic nerve: paradoxic hypertrophy after amputation in young patients. AB - PURPOSE: To evaluate the appearance of the sciatic nerve after leg amputation. MATERIALS AND METHODS: Magnetic resonance (MR) images were obtained in seven patients (age at amputation, 11-19 years) who underwent above-knee amputation to treat osteogenic sarcoma. Images were evaluated for sciatic nerve enlargement. Findings were correlated with the time after amputation. RESULTS: All seven patients were found to have a markedly enlarged sciatic nerve in the stump of the amputated leg. The enlargement extended proximally from the point of nerve transection to a level posterior to the femoral neck (8-27 cm) depending on the length of the stump. No evidence of sciatic nerve enlargement was found in the opposite leg or on preoperative MR images that were available in three of the patients. Moreover, in one patient with a sarcoma who underwent a leg-sparing procedure, no sciatic nerve enlargement was seen postoperatively. The thickness of the distal sciatic nerve was related to the time after amputation. CONCLUSION: Hypertrophy of the sciatic nerve occurred after above-knee amputation in young patients. This finding differed from atrophy of the nerve that has been reported previously in older patients. PMID- 9356644 TI - Glycosaminoglycan in articular cartilage: in vivo assessment with delayed Gd(DTPA)(2-)-enhanced MR imaging. AB - PURPOSE: To investigate the feasibility of applying magnetic resonance (MR) imaging with use of an anionic compound, Gd(DTPA)2- (gadolinium diethylenetriamine-pentaacetic acid), for measuring glycosaminoglycan concentration in human cartilage in clinical studies. MATERIALS AND METHODS: Penetration of Gd(DTPA)2- into cartilage was monitored through sequential T1 calculated images obtained after intraarticular (n = 2) or intravenous (n = 2) injection. T1-weighted and T1-calculated image series were then obtained in seven volunteers (nine knees) after penetration of Gd-(DTPA)2- into cartilage. If T1 was heterogeneous on Gd(DTPA)(2-)-enhanced images, images were also obtained after penetration of the cartilage with the nonionic contrast agent, gadoteridol. RESULTS: Gd(DTPA)2- penetrated cartilage from the articular surface after intraarticular injection and from both the articular surface and the subchondral bone after intravenous injection. The latter resulted in shorter overall penetration time. T1 values on Gd(DTPA)(2-)-enhanced images were homogeneous in four knees, but in five knees T1 differences of up to 30% were observed. These T1 differences were not seen in the presence of gadoteridol. These variations in T1 reflected about 50% variations in glycosaminoglycan. CONCLUSION: The data suggest that Gd(DTPA)(2-)-enhanced MR imaging has potential for monitoring glycosaminoglycan content of cartilage in vivo. PMID- 9356646 TI - Impaired cerebrovascular autoregulation after hypoxic-ischemic injury in extremely low-birth-weight neonates: detection with power and pulsed wave Doppler US. AB - PURPOSE: To evaluate regional cerebral blood flow with power and pulsed wave Doppler ultrasound (US) in extremely low-birth-weight neonates with periventricular leukomalacia (PVL), germinal matrix hemorrhage (GMH), or both. MATERIALS AND METHODS: The lenticulostriate arteries of 17 preterm neonates (birth weight < or = 1,100 g) were assessed daily with Doppler US during the first 5-6 days of life. The mean arterial pressure and bilateral peak velocity, resistive index, coronal vascular cross-sectional area, and product of the peak velocity and vascular cross-sectional area were measured. RESULTS: Five neonates developed PVL, GMH, or both; results of follow-up examinations in 11 patients were normal. One neonate with severe intrauterine growth retardation and renal tubular acidosis was excluded. Neonates with PVL, GMH, or both showed significantly greater mean values and more variable values of vascular cross sectional area and product of peak velocity and cross-sectional area than neonates without PVL or GMH (P < .025). Mean resistive index was significantly lower in neonates with PVL, GMH, or both than in neonates without (P < .01). There were no significant differences between mean arterial pressure in neonates with and those without PVL, GMH, or both. CONCLUSION: By enabling the detection of autoregulatory fluctuations in cerebral blood flow associated with hypoxic ischemic injury, power and pulsed wave Doppler US may enable identification of preterm neonates who are at risk of developing PVL, GMH, or both during the 1st week of life. PMID- 9356647 TI - Treatment of ectopic pregnancy: is a human chorionic gonadotropin level of 2,000 mIU/mL a reasonable threshold? AB - PURPOSE: To determine whether a human chorionic gonadotropin (hCG) level of 2,000 mIU/mL is a reasonable threshold for diagnosing ectopic pregnancy in the absence of ultrasound (US) findings of intrauterine pregnancy (IUP) and thus to prevent inappropriate treatment that will result in the loss of an otherwise normal pregnancy in women with early IUPs. MATERIALS AND METHODS: The authors reviewed the medical records of and US scans obtained in 676 patients in whom ectopic pregnancy was clinically suspected between January 1, 1994, and December 31, 1995. RESULTS: Five hundred forty-eight patients had evidence of a normal or abnormal IUP. Fifty-one (40%) of the 128 patients without evidence of an IUP had an hCG level of more than 2,000 mIU/mL. Of these 51 patients, 15 (29%) were treated for ectopic pregnancy; 17 (33%) were not immediately treated for ectopic pregnancy and had a normal IUP at follow-up US. CONCLUSION: An hCG level of 2,000 mIU/mL without US findings of IUP, while suggestive of an abnormal pregnancy, is not diagnostic. Per the results of recent studies, it is reasonable to closely follow up rather than treat many of these early, stable cases of ectopic pregnancy. PMID- 9356648 TI - Gel tubes: a method of core-specimen retrieval and transport. AB - The utility of saving core biopsy specimens in sterile gel tubes was evaluated with 30 liver and 30 kidney core specimens obtained at open biopsy with an 18 gauge needle during autopsy. The core specimens were saved with one of three methods: gel tube, formalin swirl, or scalpel retrieval. The combined-organ mean time for the radiologist to save core specimens was not statistically significantly different with gel tubes (3.8 seconds) and the formalin-swirl method (4.5 seconds). Both of these methods, however, were significantly faster than the scalpel-retrieval method (11.2 seconds) (P < .001). The combined radiologist and cytotechnologist time was greatest with the gel tubes (35.1 seconds vs 14.7 and 21.2 seconds, respectively). Core specimens were broken with the formalin-swirl and scalpel-retrieval methods but not with the gel-tube method. PMID- 9356650 TI - Extreme variability of cerebrovascular US results and some solutions. PMID- 9356649 TI - Endovascular therapy for intracranial aneurysms. PMID- 9356651 TI - Future of teleradiology. PMID- 9356652 TI - Enhancement velocity not necessarily reflective of contrast material uptake in breast MR imaging. PMID- 9356653 TI - Vascular rings and the straight trachea. PMID- 9356654 TI - Percutaneous US-guided ablation of liver metastases. PMID- 9356655 TI - Hepatitis G virus (HGV)--association with graft failure after hematopoietic stem cell transplantation? AB - A group of 28 children was investigated after hematopoietic stem cell transplantation for evidence of hepatitis G virus (HGV) infection. HGV RNA was detected in 14 of the 28 patients (50%) and persisted in 9 of 11 patients with follow-up samples for up to 32 months. Whereas thrombopoiesis was delayed in 2 of the 14 HGV-RNA-negative patients (14.3%), 6 out of 14 (42.9%) patients in the HGV RNA-positive group had a delayed thrombopoiesis and 2 of the latter group had to be retransplanted because of complete graft failure. These were the only cases with hepatitis C virus (HCV) coinfection. Significant liver diseases were also found only in these 2 patients with HGV and HCV coinfection. These results suggest that HGV infection may significantly influence the engraftment in patients after hematopoietic stem cell transplantation, particularly if coinfection with HCV occurs. PMID- 9356657 TI - Photodynamic virus inactivation of thrombocyte concentrates by phenothiazine dyes. PMID- 9356656 TI - Infection cycle of herpes viruses after photodynamic treatment with methylene blue and light. AB - Herpes simplex virus type 1 was inactivated by illumination with red light in the presence of low concentrations (1 microM) of methylene blue (MB). In ultrastructural examinations after photodynamic treatment, no significant damage of the viral envelope was observed. The inactivated viruses were able to bind to host cells and to penetrate them. Viral DNA was released within the cells, but DNA replication was completely inhibited by photodynamic treatment. In Southern blots, a degradation of the viral DNA was detected with increasing illumination time. Taken together, these data show that MB/light treatment of herpes viruses gives rise to DNA damage and blocks DNA replication. PMID- 9356658 TI - Comparative investigations of quality of erythrocyte concentrates in preservation media SAG-M, PAGGS-M and Adsol without and with leukocyte depletion. PMID- 9356659 TI - Transport of potassium ions in human red blood cells and lymphocyte survival in erythrocyte concentrates after gamma irradiation and during storage. PMID- 9356660 TI - Cryopreservation of peripheral blood progenitor cells: characteristics of suitable techniques. AB - The influence of 6 different cooling rates (1.4, 5, 10, 15, 20, 160 K/min) and 5 different compositions of the suspensions to be frozen (% DMSO/% HES: 10/0, 7.5/2.5, 5/6, 2.5/7.5, 0/10) were investigated in 30 aliquots from each of 12 peripheral blood progenitor cells (PBSC) products which had been obtained by leukapheresis from 11 patients with hematological malignancies and solid tumors and 1 healthy donor treated with 5-24 micrograms/kg body weight/day granulocyte colony stimulating factor (G-CSF) over 5 days. The MNC concentration in the products obtained amounted to 4.51 +/- 1.55 x 10(7)/ml (mean +/- SEM), range: 2.10-7.65 x 10(7)/ml). For freezing, cooling rates were generated by means of a liquid nitrogen(LN2)-operated, computer-controlled freezer, a mechanical -80 degrees C freezer, in the vapor phase over LN2, and by submerging into LN2. The statistical analysis of the results clearly indicates that optimum results compared with the prefreeze values for numerical recovery (80.6 +/- 20.1%, Coulter counter), viability (membrane integrity by Trypan blue exclusion 91.6 +/- 4.1%), and colony-forming potential (56.2 +/- 45.8% erythroid burst-forming units [BFU-E], 63.4 +/- 72.8% myeloid colonies [CFU-GM]) were achieved at a cooling rate of 1.4 K/min with 10% DMSO being present. The values obtained at a cooling rate of 5 K/min (-80 degrees C mechanical refrigerator) in the presence of 5% DMSO and 6% HES did not differ significantly (i.e., membrane integrity 91.8 +/- 3.9%, BFU-E 53.0 +/- 37.4%, CFU-GM 47.8 +/- 58.2%). At cooling rates above 5 K/min and DMSO concentrations lower than 5% (in spite of higher HES concentrations) there was a significant drop of CFU recovery (CFU-GM plus BFU-E) to almost 0%. In parallel, numerical recovery and viability dropped as well, but less pronounced, indicating that both methods are unsuitable for the prediction of CFU recovery when different freezing protocols are applied. We need at least 5% DMSO (in the presence of 6% HES) in spite of the undesirable histamine releasing effect of this compound. The cooling rate is not critical in the range from 1 to 5 K/min and can easily be achieved by -80 degrees C freezers. PMID- 9356661 TI - Leuko-reduction using an integral Sepacell filter early (4 h, 24 h) or late (48 h, 120 h) after plateletpheresis does not affect platelet function. AB - Leuco-reduction of aliquots of single-donor platelet concentrates with an integral Sepacell PLS-5A filter was performed 4, 24, and 48 h after apheresis including a control after 120 h (not recommended by the manufacturer) to evaluate the effect of both early and late filtration on the concentrates in a paired study. Highly efficient (> 2 log10) leuko-reduction was consistently observed for filtration any time after apheresis. Various proaggregatory stimuli were tested to determine the stimulus concentration (EC50 values) required for half-maximal aggregation of platelet samples (200 platelets/nl). Neither glycoprotein IIb/IIIa dependent (thrombin-, collagen-, and APD-induced) nor glycoprotein Ib-mediated (ristocetin-induced) platelet function were affected by filtration 4, 24, 48, or 120 h after plateletpheresis. For single-donor plateletpheresis using a closed system with an integral Sepacell filter, our in vitro results suggest that the timing of leuko-reduction does not affect the platelet-dependent hemostatic qualities of the product. PMID- 9356662 TI - Platelet aggregation in response to collagen and thrombin reliably detects the ingestion of low-dose aspirin. AB - The exposure of blood donors to aspirin is not reliably excluded by pharmacokinetic measurements due to the irreversible effects of aspirin which persist even after elimination of aspirin and its metabolites from plasma. Tests of platelet functions do overcome this deficit, but are usually limited by substantial inter-individual variability of parameters of platelet function. We have evaluated platelet aggregation ex vivo in 20 aspirin-treated (100 mg single oral dose/day) patients in comparison with a control group of 20 aspirin-free donors. The results demonstrate a significant reduction in the collagen(2.5 micrograms/ml)-induced platelet aggregation by aspirin treatment, whereas thrombin(50 mumol/l TRAP-6)-induced platelet aggregation was not affected at all. Assessment of collagen-induced platelet aggregation relative to platelet responses of the same subject elicited either by thrombin or by a combination of collagen and thrombin does substantially improve the reliability of functional assays of aspirin. PMID- 9356663 TI - New definition and methods for isolation of the earliest peripheral blood-derived hematopoietic stem cells. AB - Peripheral blood contains transplantable pluripotent hematopoietic progenitor cells, which are defined by the expression of certain surface antigens, such as CD34, and the absence of lineage-specific markers (lin-). Here we describe CD34- DR- adherent growing hematopoietic progenitor precursors which were separated and isolated from peripheral blood by interleukin 6(IL-6)-mediated plastic adherence. These peripheral blood mononuclear cells attach to the plastic surface of tissue culture flasks in the presence of human recombinant IL-6 and assume fibroblast like morphology. These adherent cells are CD34- and HLA-DR-, and respond to various growth factors with the expression of surface antigens, changes in morphology and the expression of cell cycle-controlling kinases. Stem cell factor induces differentiation of the CD34- DR- adherent cells to become nonadherent and to differentiate into CD34+, and eventually HLA-DR+ cells, and produce colony forming units (CFU) in semi-solid agar. While IL-6 conveys adherence of these cells, the addition of stem cell factor induces differentiation into nonadherent cells. Our observations suggest the possibility of isolating a true hematopoietic stem cell before the level of CD34 and HLA-DR expression. PMID- 9356664 TI - Haematopoietic reconstitution after autologous transplantation of CD34(+) selected versus non-selected peripheral blood progenitor cells. AB - Selection of CD34+ cells for autologous transplantation is increasingly being used to reduce potential tumor cell contamination of the autograft. Haematopoietic reconstitution in 40 patients after transplantation of CD34(+) selected versus non-selected G-CSF-mobilized PBPC was compared and was almost identical in the two groups of patients. Delayed platelet engraftment was only observed in patients transplanted with a CD34+ cell dose of < 2.5 x 10(6)/kg body weight. It has to be shown whether the positive selection of CD34+ cells will improve the disease free survival after autologous PBPC transplantation. PMID- 9356665 TI - High efficiency of CD34+ selection by removal of platelets from the apheresis product. PMID- 9356666 TI - Rapid donor type isoagglutinin production after allogeneic peripheral progenitor cell transplantation. AB - Signs of haemolysis and donor-type isoagglutinin production were observed in a patient with minor ABO incompatibility following peripheral progenitor cell transplantation with posttransplant cyclosporine for GvHD prophylaxis. Renal failure was avoided by immediate forced diuresis. PMID- 9356667 TI - The influence of extracorporeal peripheral blood stem cell apheresis on platelet antigens. PMID- 9356668 TI - Detection of malignant cells of epithelial origin in mononuclear cell fractions. AB - Qualitative and quantitative detection of contaminating malignant cells in stem cell apheresis products intended for supportive therapy after high-dose chemotherapy of sensitive solid tumors is a demanding task for the laboratory. Human breast cancer cells in differing concentrations among mononuclear cells were quantitated by multicolor flow cytometry and immunocytochemistry. Comparing specificity, sensitivity, and laboratory processing time, the immunocytochemical detection of the above-mentioned malignant cells of epithelial origin was demonstrated to be the superior method. PMID- 9356669 TI - Positive selection of CD34+ cells by immune affinity. PMID- 9356670 TI - Identification of IgG alloantibodies in patients with high-titer IgM cold agglutinins by serum/plasma affinity chromatography. AB - The detection of IgG alloantibodies in the presence of high-titer cold autoagglutinins (CAs) can be extremely difficult, especially under pressure of time when transfusion of red blood cells is urgently needed. Here we demonstrate that IgG alloantibodies in the presence of high-titer IgM CAs can be easily detected by quantitative IgG purification from serum or plasma by affinity chromatography. In comparison with the routinely used methods for IgG alloantibody identification, affinity chromatography shows better or identical results and is the method leading to results most rapidly. PMID- 9356671 TI - CMV-induced anti-Sia-b1 cold agglutinin in an immunocompromised patient. AB - In postinfection cold agglutination, certain cold agglutinin (CA) specificities are associated with distinct infectious agents. The combined occurrence of anti-I and anti-Sia-b1 CAs following Mycoplasma pneumoniae infection has been reported recently. After renal transplantation and hyperacute graft rejection, transiently occurring CAs were observed in an 18-year-old boy. The CAs were characterized by serum cold absorption with sialidase-treated red cells and warm elution from the cells. An anti-Sia-b1 CA could be differentiated from an accompanying low-liter anti-I. Fresh infections with Mycoplasma pneumoniae, Epstein-Barr virus, rubella, and varicella viruses were excluded, but CMV infection was demonstrated. This is the first case of a postinfection anti-Sia-b1 CA associated with CMV infection. PMID- 9356672 TI - Case report: anti-Coa in a Co-(a+)-typed patient with chronic renal insufficiency. AB - The recently identified molecular structure of the Colton blood group system is characterized by an amino acid substitution at position 45 (Colton a: alanine, Colton b: valine) of the archetypical water channel protein Aquaporin-1 (AQP1), which regulates water homeostasis in the erythrocyte membrane and in the proximal tubule of the nephron. We identified a patient with the unique constellation of an antibody with the specificity anti-Coa and the Co (a+) phenotype. The serological antigen typing was confirmed by molecular typing with PCR-RFLP. The antibody has to be interpreted as an antibody against a partial Colton a antigen or as an autoantibody despite a negative direct antiglobulin test (DAT). The patient is suffering from chronic renal insufficiency of unknown origin, rising speculation about a pathophysiological relationship between the serological constellation and the clinical disease under the aspect of localization of the Colton antigens on AQP1. PMID- 9356674 TI - On the formation of epitopes in the Rh system. PMID- 9356673 TI - Limits of the MAIPA assay when differentiating high-titered platelet-reactive antibodies. AB - Experience with the MAIPA assay for the diagnosis of platelet-reactive antibodies has shown that high-titered antibodies falsify the test results. We here demonstrate 2 cases: i) A serum with high-titered HLA antibodies (100% panel reactivity in the LCT, titer between 4,000 and 12,000), and ii) Serum with a high titered anti-HPA-1a (titer in the MAIPA assay 1,000). In both cases, it can be demonstrated that these antibodies led to unspecific reactions. In the 1st case, they interfered with the diagnosis of additional platelet-specific antibodies. Only the use of HLA-compatible platelets allowed a correct identification. On the other hand, in the high-titered anti-HPA-1a unspecific reactions were seen with the glycoproteins Ib/IX, Ia/IIa, and beta 2-microglobulin, leading to misinterpretations. These examples demonstrate that, in the test conditions as described, a correct diagnosis of high-titered sera might only be achieved by using compatible HLA or HPA cells. PMID- 9356675 TI - Routine rhesus-D genotyping in amniotic fluid: analysis of exon 10 versus intron 4 by polymerase chain reaction. AB - Both the exon 10 and the intron between exons 4 and 5 of the Rh-D gene were analyzed in DNA from amniotic cells by polymerase chain reaction (PCR) in order to determine the fetal Rh-D status. Residual (0.3-2 ml) samples of amniotic fluid obtained by diagnostic amniocentesis in Rh-D-negative women were analyzed. It is important to standardize the sampling and preanalytical storage and to optimize DNA extraction procedures as well as the detection of PCR products for maximal sensitivity. Genotyping was successful in approximately 90% (229 of 256) of amniotic fluid samples tested. Consistent Rh-D results were obtained by both methods in all 229 typed samples with a single exception. Our experience demonstrates that routine genotyping of the Rh-D blood group in small volumes of amniotic fluid (0.5-2 ml) is feasible. Its validity is currently tested by comparison of the prenatal genotype with the serotype of the newborn. PMID- 9356676 TI - Enhancement of Sia-lb1 antigenicity by sulphur-containing substituents at C-5 of free N-acetylneuraminic acid. AB - The Sia-lb1 epitope, recognized by anti-Sia-lb1 cold agglutinins, is unique since it is represented by the alpha-N-acetylneuraminic acid (alpha NeuNAc) monosaccharide. Chemical modifications of the chain at C-5 of alpha NeuNAc have shown that the natural 2-carbon and the artificial 3-carbon chains are optimal for anti-Sia-lb1 binding. Sia-lb1 antigenicity of alpha NeuNAc could be tenfold enhanced by replacement of the carbonyl oxygen by sulphur. The structural requirements of the Sia-lb1 epitope for optimal antibody binding were identified. PMID- 9356677 TI - Mutations in severe hemophilia A: distribution within the factor VIII gene, origin and influence on inhibitor development. PMID- 9356678 TI - The diversity of the HLA class I introns reflects the serological relationship of the coding regions. AB - The introduction of PCR-based HLA typing techniques has uncovered that the HLA system is much more variable than it has been expected from the conventional typing methods. More and more new alleles are detected which are not characterized by new sequence motifs, but by new combinations of already existing sequence motifs. This variability, reflecting the immunological necessity to have the greatest capacity for presenting antigenic peptides, is increasingly complicating DNA typing techniques based on the diversity of the coding region. We have determined the sequence of the 1st through 3rd intron of the majority of HLA-A and HLA-B alleles in 48 well-defined cell lines and 195 PCR-typed clinical samples. The few published sequences emerged to contain substantial errors. The introns turned out to be highly polymorphic. Besides extensive homologies, numerous locus- and group-specific sites could be identified. The most intriguing finding was that most of the polymorphic motifs were related to serological families. These sequence motifs were extremely beneficial for setting up PCR based typing systems. In particular, sequencing-based typing strategies benefited from intron-restricted priming for amplification and sequencing by enabling complete analysis of the polymorphic exons. The determination of cis/trans linkages of sequence motifs was substantially facilitated. Apart from these advantages, the intron sequences were useful for evolutionary studies, delivering more insights into the genetic relationship between different alleles and the mechanisms involved in the development of the diversity of HLA. Moreover, the variability of the introns may provide a structural basis for the identification of regulatory elements acting on the level of transcription. PMID- 9356679 TI - Allele-specific PCR amplification of factor V Leiden to identify patients at risk for thromboembolism. AB - Resistance to activated protein C is the most common hereditary cause of thrombophilia and is significantly linked to factor V Leiden. We designed primers in order to identify factor V Leiden by allele-specific PCR amplification. Amplification specificity for factor V was ensured by a 3' primer located at the intron 9/exon 10 border of the gene. One sense and two antisense primers were used in two separate primer mixes specific for factor V ARG506 (wild-type) or factor V GLN506 (factor V Leiden). In each PCR reaction a pair of primers amplifying a fragment of the human growth hormone gene was included as an internal positive amplification control. The presence or absence of specific PCR amplification allowed definite allele assignment without the need for any postamplification specificity step. The internal positive control primers indicate a successful PCR amplification, allowing the assignment of homozygosity. In a prospective study 126 patients with thromboembolic events were analyzed using this technique and PCR-RFLP. The concordance between these methods was 100%. In 27 patients a heterozygous factor V GLN506 mutation was detected, whereas 1 patient with recurrent thromboembolism was homozygous. No false positive or false-negative results were observed in the homozygous as well as heterozygous samples. Additionally, in 15 samples identified to carry the point mutation by allele-specific PCR amplification, automatic sequencing has confirmed the heterozygous or homozygous point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor V Leiden. PMID- 9356681 TI - Proceedings of workshop: status of peritoneal dialysis in the United States. National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. PMID- 9356680 TI - Effects of standard unfractionated and low-molecular-weight heparins on platelets of patients undergoing total hip replacement surgery. AB - For prevention of venous thromboembolism in general and in orthopedic surgery, patients are treated prophylactically with standard unfractionated heparins (SH) or with low-molecular-weight heparins (LMWH). Patients (n = 22) undergoing total hip replacement surgery received either 5,000 IU SH (Heparin-Na) three times per day (n = 10 patients) or LMWH (Fraxiparin) once per day (n = 12 patients). Blood samples using CTAD (citrate, theophylline, adenine, dipyridamole) as anticoagulant were collected perioperatively after the first heparin administration, after the operation, and daily until day 4 postoperatively. After cooling at 4 degrees C blood samples were centrifuged, platelet-rich plasmas (PRPs) prepared, and after platelet counting PRPs were divided into platelet sediments (PS) and platelet-poor plasmas (PPP). Cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), serotonin, P-selectin, and laminin were analyzed in PPP and in PS after two freezing/thawing cycles. Platelet serotonin contents and serotonin release did not differ in either heparin group (SH and LMWH). In the LMWH group, 72 h postoperatively the intraplatelet cAMP was significantly (p < 0.01) higher. P-selectin values in PPP and platelet P-selectin release did not vary between the SH and the LMWH group; on the contrary, the platelet P-selectin content increased in the LMWH group 72 and 96 h postoperatively, while in the SH group this parameter showed a small decrease. The differences were significant (p < 0.05). The platelet-bound laminin underwent a slight change 48 and 72 h postoperatively in the LMWH group, but in the SH group the platelet-bound laminin increased permanently and significantly (p < 0.05; p < 0.01) until 72 h postoperatively, but 96 h postoperatively there was a small decline. In the SH group, 24-96 h postoperatively the platelet-bound laminin was significantly (p < 0.05; p < 0.005) augmented, compared with the LMWH group. The higher cAMP and P-selectin contents in the LMWH group suggest that platelets are less impaired by LMWH, and the augmented platelet-bound laminin in the SH group could express the platelet impairment evoked by standard unfractionated heparins. PMID- 9356682 TI - End-stage renal disease, managed care, and capitation: implications for the renal community. PMID- 9356683 TI - Some of the small print on managed care proposals for end-stage renal disease. AB - In this article we discuss selected issues related to Medicare's end-stage renal disease (ESRD) managed care demonstration project and Congressional proposals to remove the barrier to ESRD patients enrolling in Medicare managed care plans. We discuss financial incentives to keep patients healthy; beneficiary obligations under fee-for-service and managed care; risk selection by beneficiaries among plans; and the baseline determination of a capitation rate. The ESRD demonstration offers the opportunity to evaluate the consequences of making Medicare managed care options available to a high cost and clinically vulnerable population. Careful evaluation is necessary to ensure that ESRD managed care options are structured to be beneficial to taxpayers, caregivers, and, most importantly, the beneficiaries choosing these options. Certainly, the potential exists for managed care to benefit patients by changing the fractured system in which each provider only has an incentive to worry about its own costs. However, the possible unintended consequences highlighted in this article strongly suggest that the evaluation of the demonstration project be undertaken before managed care options are made widely available outside the demonstration sites. Problems of a more technical nature, such as how to best use available Health Care Financing Administration data in the rate-setting process, are likely to be overcome, but the time and effort necessary to resolve them should not be underestimated. PMID- 9356684 TI - Capitation in nephrology: the time has come. AB - Nephrology represents a powerful example of the impact of new medical technology; almost 200,000 patients are alive in the United States because of renal replacement therapy. However, ESRD patient morbidity and life expectancy are still serious concerns despite advances in medical treatment. A capitated payment system for end-stage renal disease (ESRD) could be a great benefit to the renal community as it would force us to rethink the way ESRD care is provided. By applying the concepts of integration of services, disease management, and case management to ESRD care under a capitated payment system, providers may be able to improve patient outcomes and overall well-being. PMID- 9356685 TI - Development of an end-stage renal disease managed care demonstration. AB - At present, end-stage renal disease (ESRD) beneficiaries cannot enroll in health maintenance organizations (HMOs) or social health maintenance organizations (SHMOs), but HMO members who develop ESRD may remain enrolled, and the Health Care Financing Administration (HCFA) pays the HMO a state-specific, but otherwise unadjusted, capitation rate that is 95% of fee-for-service (FFS) costs. Thus, more than 6,000 ESRD beneficiaries were enrolled in HMOs in 1993, when Congress mandated an ESRD SHMO demonstration in which not only Medicare-covered services, but extra benefits were to be provided to Medicare beneficiaries, with the SHMO receiving a capitation rate based on 100% of FFS costs. The demonstration will test (1) the feasibility of year-round open enrollment of ESRD beneficiaries in HMOs; (2) a capitation system based on treatment status--dialysis, transplant, or functioning graft--and adjusted for age and whether diabetes was the cause of renal failure; (3) the effect of the additional benefits; and (4) whether managed care can improve ESRD quality outcomes. HCFA made demonstration awards in September 1996 to Kaiser-Permanente in Southern California; Health Options in Southern Florida; and Phoenix Healthcare in Central Tennessee. The sites are expected to have 1 year of planning and development before beginning the congressionally mandated 3 years of service delivery. There will be an independent evaluation. PMID- 9356686 TI - The institution of care pathways in nephrology patient care: a response to the changing health care climate. AB - The development of managed health care in the United States has provided an impetus for new strategies that promote efficiency, streamline healthcare delivery, and maintain quality care. The increasing number of end-stage renal disease patients, their complexity of care, and a looming manpower shortage in nephrology strain the present system trying to meet these demands. One mode of healthcare delivery that may address specific needs in the nephrology population is case management. This approach to medical care uses a care pathway that serves as a multidisciplinary blueprint for patient care. Such pathways eliminate duplicated services and maximize efficiency by keeping the healthcare team focused. In response to market forces in our community, we implemented care pathways for percutaneous renal biopsy and vascular access surgery. Costs per procedure and hospital length of stay were reduced. Patient outcomes and procedure success rates were unchanged from pre-pathway years. Moreover, patients preferred the care pathway care for their problems. Case management and care pathways are tools that are effective in their scope for helping deliver better care for nephrology patients. While they should not be considered a panacea for the problems facing renal care providers, these tools should be considered as part of nephrology healthcare delivery in the future. PMID- 9356687 TI - The relationship between quality of care and financial performance in dialysis: "doing well by doing good". AB - Dialysis is an archetype for the application of the principles of continuous quality improvement (CQI) to medicine. Dialysis treatment dosage and hematocrit values are just two examples of data that can be readily obtained for reliable and valid quality analysis and improvement of the dialysis process. A dialysis CQI program should focus on the improvement of care processes to effect improvements in patient outcomes and should function through the teamwork of caregivers at all levels within an organization. By succeeding in the continuous improvement of the multiple processes involved in the delivery of chronic dialysis to patients, dialysis providers can expect to see improvement in patient outcomes and, as a direct result, improvement in financial performance, both in the fee-for-service, as well as in the capitated model of reimbursement. PMID- 9356688 TI - The use of mediation to manage patient-staff conflict in the dialysis clinic. AB - The use of mediation to manage a disruptive incident between a staff nurse and a hemodialysis patient is the focus of this multidisciplinary case review. The growing interest in techniques to manage disruptive patient behavior in the dialysis clinic is analyzed. A mediation model designed for the dialysis clinic is presented for the reader. This case presentation also examines the experience of conflict from several perspectives including the patient, the nurse, the nurse manager, and the social worker. This discussion is intended to provide the reader with support for viewing conflict as unavoidable in the chronic dialysis treatment environment and for enlisting the support of the patient when responding to disruption in the treatment relationship. PMID- 9356689 TI - Mediation for challenging patients--a promising approach. PMID- 9356690 TI - Central venous dialysis catheter dysfunction. AB - Central venous catheter dysfunction is a limiting factor in regard to renal replacement therapy efficiency and can thus influence patient morbidity. Early catheter dysfunction is frequently due to mechanical problems such as inadequate positioning, kinking, or constriction, but early fibrin deposition can develop soon after insertion. Delayed dysfunction usually results from thrombus formation, either within the lumen, around the catheter ("fibrin sleeve"), or in the host vein. Catheter dysfunction is suspected clinically or documented by simple imaging studies. It is usually evident and manifested by failure to aspirate blood from the lumen(s), inadequate blood flow and/or high resistance pressures during hemodialysis. However, a more subtle dysfunction may lead to a high recirculation of dialyzed blood and be overlooked if dialysis adequacy is not monitored regularly. Local instillation of a fibrinolytic agent is usually successful in restoring catheter patency. Central venous dialysis catheters present intrinsic limitations consequent to their composition and design, whereas extrinsic limitations result from site of insertion, blood properties and anatomic particularities of a given individual. These characteristics largely determine overall catheter performances. Performance parameters to consider include maximal consistently achievable blood flow rate, resistance to blood flow indicated by arterial and venous pressures during hemodialysis, and blood recirculation rate. Catheter longevity is an important consideration for cuffed catheters implanted for long-term use. The tolerated blood recirculation within central venous dialysis catheters should be below 10% to 15%, and is ideally between 3% to 7% in most clinical settings. Several recent studies confirm that short femoral catheters recirculate significantly more than is desirable. Well functioning and nonreversed internal jugular and subclavian venous catheters have, in general, recirculation rates less than 5%. With regard to various performance criteria, the TwinCath (Medcomp, Harleysville, PA) appears particularly advantageous. In any case, a good catheter maintenance program is of critical importance for the prevention and the early detection of catheter dysfunction. PMID- 9356691 TI - Wound care management in the end-stage renal disease population. AB - As the end-stage renal disease patient population ages, the management of pressure ulcers and vascular ulcers becomes an increasingly common problem. The practice of wound care management and the devices used to manage wounds has changed dramatically. Basic knowledge of the healing process and new techniques in wound care is paramount for the nephrology team. Identification of patients at risk for wounds and the development of wound care models is one step in the goal towards preventative care. PMID- 9356692 TI - Collaborative practice of renal nutrition in end-stage renal disease patient care. PMID- 9356693 TI - My kidney failure experiences. PMID- 9356694 TI - Effect of cytoxan-induced heteropenia on the response of specific-pathogen-free chickens to infectious bronchitis. AB - Epithelial damage in infectious bronchitis occurs early in the disease process. Heterophil infiltration into the tracheal mucosa is greatest at that time. To determine the contribution of heterophils to tracheal epithelial damage of infectious bronchitis, eight 3-wk-old specific-pathogen-free chickens were made heteropenic by four daily intramuscular injections of cyclophosphamide at 75 mg/kg body weight. Infection with Massachusetts 41 infectious bronchitis virus was timed to coordinate heteropenia with peak tracheal epithelial damage. Heteropenia was monitored by total leukocyte and differential cell counts of peripheral blood. Tissue damage and heterophil infiltrate were monitored by histopathology of tissues taken at termination of the study. Heteropenic birds had lower peripheral blood and tracheal heterophil numbers than nonheteropenic birds. No difference was found in epithelial damage of heteropenic and nonheteropenic birds. Epithelial damage in infectious bronchitis is most likely due to damage by the virus and not due to the infiltrated heterophils. PMID- 9356696 TI - A simple immunoperoxidase plaque assay to detect and quantitate Marek's disease virus plaques. AB - We report an immunoperoxidase-based staining technique that can be used to rapidly and accurately detect and quantitate Marek's disease virus (MDV) plaques. Monolayer cultures were fixed and incubated with a monoclonal antibody specific for MDV. After washing, a second antibody of horseradish peroxidase-conjugated goat anti-mouse IgG was applied, incubated for 1 hr, and washed with phosphate buffered saline. After the cultures were incubated with diaminobenzidine, CoCl2, and H2O2, the plaques appeared as black spots and were easily seen and counted. Significantly more immunoperoxidase-stained serotype 1 MDV plaques could be counted at 4 days postinoculation than were seen in unstained cultures. With serotype 2 MDV-infected cells, the difference in plaque counts was less dramatic. Nevertheless, at 3 days postinoculation, significantly more stained serotype 2 plaques were seen than unstained plaques. Immunoperoxidase staining of turkey herpesvirus plaques did not increase the sensitivity of viewing plaques. Similar numbers of stained and unstained plaques were seen at 2 days postinoculation. We also demonstrated that we could count serotype-specific MDV plaques in a mixed infection that contained all three serotypes. PMID- 9356695 TI - Systemic and local antibody responses to infectious bronchitis virus in chickens inoculated with infectious bursal disease virus and control chickens. AB - Serum and local (respiratory) antibody responses to infectious bronchitis virus (IBV) were studied in 5-wk-old white leghorn-type control chickens and chickens inoculated with infectious bursal disease virus (IBDV) at 1 day of age. Of the chickens inoculated with IBV alone, 93% had detectable levels of IBV antibodies in the sera and 87% had detectable antibodies in the respiratory lavage fluids. Compared to this group, only 73% and 65% of IBDV-IBV inoculated chickens had serum and respiratory antibodies, respectively. In chickens inoculated with IBV alone, the IBV antibodies were evenly associated with immunoglobulin classes IgM, IgG, and IgA, whereas the IBDV-IBV inoculated chickens mainly produced IgM associated antibodies with low to negligible levels of IgA- and IgG-associated antibodies. These results suggest that the lack of adequate IgA- and IgG associated antibody production in IBDV-infected chickens may account for their increased susceptibility to IBV infection. PMID- 9356697 TI - Invasion of chicken reproductive tissues and forming eggs is not unique to Salmonella enteritidis. AB - Experiments were conducted in which Salmonella enteritidis Phage Type 8, Phage Type 2, and RDNC (reaction does not conform) or three isolates of Salmonella typhimurium of diverse origin were fed to adult laying hens to determine if S. enteritidis has a selective advantage over S. typhimurium, which is now rarely isolated from chicken eggs, in its capacity to invade reproductive tissues. The results revealed that S. enteritidis and S. typhimurium may be equal in their potential to colonize the tissues of the reproductive tract and eggs that are forming in the oviduct prior to oviposition. S. enteritidis, but not S. typhimurium, was isolated from egg contents after oviposition. The degree to which intestinal, hepatic, splenic, or reproductive tissues were colonized by either serotype was not seen to affect the rate of colonization of eggs forming in the oviduct or the contamination of eggs after oviposition. Virulence factors related to the difference in the association of S. enteritidis and S. typhimurium with egg-borne salmonellosis remain to be defined. PMID- 9356698 TI - Various blood parameters in commercial hens acutely and chronically infected with Mycoplasma gallisepticum and Mycoplasma synoviae. AB - Two trials were conducted to study the effects of acute (Trial 1) and chronic (Trial 2) mycoplasma infections on differential leukocyte counts in chickens. The trials initially included either 20 (Trial 1) or 40 (Trial 2) 6-wk-old commercial leghorn chickens negative for antibodies to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). Chickens were inoculated with F strain MG (FMG), MS (WVU 1853), or both. One group of chickens remained uninoculated and served as a negative control for both trials. Chickens were housed in fiberglass isolation units from 6 to 10 wk (Trial 1) or 6 to 70 wk of age (Trial 2). Differential leukocyte counts were examined from 6 to 10 wk (Trial 1) or 66 to 70 wk of age (Trial 2) in all chickens. Also, in Trial 2, packed cell volumes (PCVs) and plasma protein values were examined from 66 to 70 wk of age. In the acute study (Trial 1), differential leukocyte counts revealed statistically significant differences (P < 0.05) in heterophil, lymphocyte, monocyte, eosinophil, and basophil values among treatments. In general, the differential counts of FMG- and MS-infected birds were characterized by heterophilia, lymphopenia, monocytosis, eosinopenia, and basopenia. Histopathologic examination of the spleen, liver, kidney, and bone marrow revealed a high degree of lymphoid foci within the spleen and bone marrow of all infected chickens. In the chronic study (Trial 2), no statistically significant differences (P < 0.05) were observed in differential leukocyte counts, PCV, and plasma protein values among treatments. Histopathologic examination of spleen, liver, kidney, and bone marrow did not reveal any difference among treatments. PMID- 9356699 TI - Inhibition of Salmonella typhimurium attachment to chicken cecal mucus by intestinal isolates of Enterobacteriaceae and lactobacilli. AB - The ability of selected strains of Enterobacteriaceae or lactobacilli isolated from the intestines of adult chickens to inhibit in vitro attachment of Salmonella typhimurium 3333/O to cecal mucus in the presence or absence of D mannose was determined. Attachment in the absence of mannose was reduced by prior exposure of mucus to cultures of two isolates of Enterobacteriaceae, an Escherichia coli and a Hafnia alvei strain, but not to a third isolate, an Enterobacter agglomerans strain. Attachment of S. typhimurium was not inhibited when mannose was present in the blocking or attachment step. Formation of fimbriae by the two inhibitory Enterobacteriaceae strains and the S. typhimurium strain, as indicated by titers of mannose-sensitive hemagglutination of guinea pig erythrocytes was optimal in Z biphasic medium (consisting of tryptone, yeast extract, dextrose, and NaCl) incubated anaerobically at 42 C. Fimbriae of each of three strains prepared from these cultures also inhibited attachment. These are characteristics consistent with attachment and inhibition of attachment mediated by a mannose-sensitive adhesin associated with type 1 fimbriae on bacterial cells of Enterobacteriaceae strains. Attachment in the presence of mannose was significantly reduced by prior exposure of mucus to cultures of a Lactobacillus salivarius strain and a Lactobacillus delbrueckii delbrueckii strain but not to a strain of Lactobacillus for which the species had not been determined. Washed cells or spent culture supernatant fluid from brain-heart infusion broth, Z broth, or Z biphasic cultures of the inhibitory strains of lactobacilli incubated at 37 or 42 C inhibited this form of attachment. Of 27 intestinal isolates of Enterobacteriaceae and 21 of lactobacilli, the lactobacilli strains were generally more hydrophobic than the Enterobacteriaceae as determined by adherence to hexadecane. The lactobacilli isolates did not agglutinate guinea pig erythrocytes. The data suggest more than one mechanism for mediating attachment of inhibitory bacterial strains and for subsequent attachment of S. typhimurium. PMID- 9356700 TI - Differences in frequency, level, and duration of cecal carriage between four outbred chicken lines infected orally with Salmonella enteritidis. AB - Four chicken lines, L2, B13, PA12 (egg-type), and Y11 (meat-type), were tested for experimental carrier state of Salmonella enteritidis (SE) in two identical trials. After oral inoculation of SE at 1 wk of age with 5 x 10(4) SE colony forming units (CFU), 10 chickens per line were necropsied weekly for 6 wk and then every 8 or 15 days until the 12th week postinoculation (PI). Liver, spleen, ovary, and ceca were examined for level of SE colonization. Numbers of positive livers and spleens and levels of the challenge strain in these organs differed little between the four chicken lines. Only three positive ovaries were detected. According to the chicken line, ceca exhibited generally significant (P < 0.05) differences in the number of positive organs during weeks 5-11 PI, in the SE CFU levels (P < 0.05) in the first 5 wk PI and during weeks 8 and 10 PI, and in the duration of colonization. L2 and B13 chickens generally carried SE in their ceca at higher levels, in more animals, and for a longer time than PA12 and Y11 chickens. Y11 chickens were the most resistant to SE cecal colonization. PMID- 9356701 TI - Colicin V38 and microcin C38 produced by Escherichia coli strain 38. AB - Escherichia coli strain 38, an isolate from turkeys, has been previously shown to produce one or more broad-spectrum bacteriocins against other related enteric bacteria. Using a collection of E. coli strains that synthesized well characterized colicins or microcins, along with a set of colicin/microcin insensitive mutants, we were able to classify the bacteriocins produced by strain 38. We determined that strain 38 produced a microcin (microcin C38) and a colicin (colicin V38) and that the amount of microcin C38 depended on the type of media on which it was grown. Escherichia coli strain 38 was found to have cross immunity with the microcin C7-producing strain MC4100 and with the colicin V producing strain 4674. OmpF mutant cells were found to be insensitive to microcin C38, whereas colicin V38 was not active on cells that had a cir mutation. Both microcin C38 and colicin V38 were inactivated by proteases. Microcin C38 was stable at extremes of pH (pH 1.5 and pH 13) and heat (10 min at 98 C) conditions, whereas colicin V38 was not. In addition, microcin C38 was found to be active against a broader spectrum of gram-negative bacteria than was colicin V38. PMID- 9356702 TI - Experimental infection of turkey poults with western equine encephalitis virus. AB - The pathogenicity of a field isolate of western equine encephalitis (WEE) virus, which was recovered from a breeder hen during investigations of egg production drops in California turkey flocks, was tested in 2-wk-old turkey poults. No symptoms or mortality were observed in poults inoculated intramuscularly with 4.2 log10 50% embryo lethal doses of virus; however, the infection did result in mild to moderate lymphoid necrosis in the bursa of Fabricius and thymus glands beginning on the first day postinoculation. In addition, WEE virus could be isolated from the blood of infected poults for up to 3 days postinoculation. PMID- 9356703 TI - Antigenic characterization of a turkey coronavirus identified in poult enteritis- and mortality syndrome-affected turkeys. AB - A turkey coronavirus (TCV [NC95]) was characterized by antigenic comparison with other avian and mammalian coronaviruses using immunofluorescence (FA) and immunoperoxidase (IP) procedures. Based on FA and IP procedures, TCV (NC95) was determined to be antigenically indistinguishable from turkey enteric (bluecomb) coronavirus (TECV). In addition, TCV (NC95) and TECV were found to be closely related to infectious bronchitis virus (IBV); a one-way antigenic relationship was demonstrated. Polyclonal antibodies specific for TECV and IBV reacted strongly against TCV (NC95), as determined by FA procedures. Monoclonal antibodies (MAbs) specific for IBV matrix protein (MAb 919) reacted strongly against TCV (NC95) and TECV as determined by FA and IP procedures; an IBV peplomer protein-specific MAb (MAb 94) did not recognize the two viruses. These studies suggest an identification of TCV (NC95) as a strain of TECV, and provide evidence of a close antigenic relationship between these viruses and IBV. PMID- 9356704 TI - Newcastle disease oil emulsion vaccines prepared with animal, vegetable, and synthetic oils. AB - Animal, vegetable, and synthetic oils were tested as potential replacements for mineral oil in Newcastle disease oil emulsion vaccines. Emulsifying surfactants of seed oil origin comprised 10% of the the oil phase that was used to prepare water-in-oil emulsion vaccines that contained a final concentration of 20% aqueous antigen. The hemagglutination inhibition responses of chickens inoculated with 46 of the newly formulated oil vaccines were, in most cases, not significantly different from those of control chickens inoculated with mineral oil vaccine. Tissue reactions associated with animal, vegetable, and synthetic oil vaccines were less severe than those associated with mineral oil vaccines. Viscosity of the mineral oil formulations ranged from 1/2 to 3 1/2 times that of the mineral oil control vaccines. These findings indicate that any of several oils may be more suitable than mineral oil for preparation of immune adjuvants for poultry vaccines. PMID- 9356705 TI - Pathogenicity of attenuated infectious bronchitis viruses for oviducts of chickens exposed in ovo. AB - A fixed effects, completely randomized factorial design was used to study the effect of infectious bronchitis virus (IBV) inoculation at two different exposure ages and three postinoculation (PI) durations on chick oviduct pathology. Maternal antibody-positive chicken embryos at 18 days of embryonation (ED) and newly hatched chicks were inoculated with an IBV vaccine (V-IBV) or with an IBV vaccine that had been serially passaged 21 times in chick kidney tissue culture (P-IBV). Hatchability of eggs inoculated with V-IBV at 18 ED was significantly lower (27%) than eggs that were not inoculated with IBV or were inoculated with P IBV (45-58%, P < 0.01). Chicks from all treatment groups survived to 5 days after hatch. Pathologic changes in the oviduct were evaluated at 9, 18, and 27 days PI by light microscopy. Inoculation of V-IBV and P-IBV in the presence of maternal antibodies did not result in any oviduct pathology at 9, 18, and 27 days PI. Respiratory clinical signs, however, were observed in 61% and 5% of chicks inoculated with V-IBV at 18 ED and at hatch, respectively. Respiratory clinical signs were not observed in control birds, birds inoculated with P-IBV at 18 ED, or birds inoculated with P-IBV at hatch. PMID- 9356706 TI - Spiking mortality of turkey poults: 1. Experimental reproduction in isolation facilities. AB - Spiking mortality of turkeys (SMT) is an infectious disease of 5-to-25-day-old turkey poults characterized by acute enteritis and bursal and thymic atrophy. Brooding 1-day-old poults on litter taken from naturally occurring cases successfully reproduced SMT 5 days postexposure. Oral exposure to an organ homogenate made of tissue samples from naturally occurring cases successfully reproduced SMT 5 days postinoculation. Coronaviruses were present in intestinal and bursal contents taken from poults with naturally occurring SMT. They were also present 5 days after exposure in the experimentally reproduced disease. Severe intestinal villus atrophy, bursal follicular lymphoid depletion, and thymic cortical atrophy were present histologically in naturally occurring SMT and in SMT reproduced by either experimental method. PMID- 9356707 TI - Identification and partial characterization of Arkansas isolates of chicken anemia virus. AB - Chickens from both broiler and broiler breeder pullet flocks experiencing symptoms of chicken anemia virus (CAV) infection were first observed at the Poultry Health Research Laboratory at the University of Arkansas in September 1992. Flocks had experienced higher than normal mortality with subcutaneous hemorrhages on the wings, neck, and thorax. Postmortem and histopathologic evaluation revealed thymus and bursal atrophy and lesions consistent with those reported for CAV infection. Because this infection had not previously been observed by Poultry Health Research Laboratory personnel in Arkansas-grown chickens, the establishment of a definitive diagnosis was deemed important. The presence of CAV was established by infecting MSB-1 cells with pooled liver homogenates from groups of 10 specific-pathogen-free chickens that had previously been inoculated in an attempt to experimentally reproduce the disease observed in the field. Cytopathic effects in the infected MSB-1 cells were first evident following the fifth passage. Indirect fluorescent antibody technique identified infected MSB-1 cells following at least five blind passages. To further confirm the presence of CAV, a polymerase chain reaction (PCR) technique was used to amplify a specific portion of the virus genome from infected MSB-1 cells and tissue extracts from several submitted chickens. Sequence analysis of a 186-bp PCR amplification product revealed that the Arkansas isolate was very similar to the Cuxhaven-1 isolate (99.5% sequence identity). PMID- 9356708 TI - Expression of MD infectious bursal disease viral proteins in baculovirus. AB - Genomic segment A of the variant infectious bursal disease virus (IBDV) strain MD was amplified by reverse transcriptase/polymerase chain reaction and further characterized by baculovirus expression. Three different baculovirus clones were constructed containing the genes encoding VP2, VP2/VP4, and the complete polyprotein cloned into the baculovirus transfer vector pVL1392. Baculovirus recombinants were identified by dot blot hybridization and were plaque purified three times. Baculovirus expression of the recombinants produced IBDV-specific proteins that were comparable in molecular size to native MD IBDV viral proteins VPX (48 kD), VP2 (45 kD), VP3 (32 kD), and VP4 (28 kD) as determined by sodium dodecyl sulfare-polyacrylamide gel electrophoresis and western immunoblot analysis. All three recombinants produced a 48-kD protein that possibly represents VPX, the precursor product of VP2. In addition to the 48-kD protein, the VP2/VP4 recombinant produced an IBDV-specific protein corresponding to the 28 kD VP4. The baculovirus-expressed polyprotein gene produced, in addition to the 48-kD protein, a 32-kD (VP3) IBDV-specific protein and a 29-kD protein that migrated slightly slower than MD VP4. The baculovirus-expressed proteins were used as antigens in an indirect enzyme-linked immunosorbent assay (ELISA). The ELISAs detected antibodies against the variant IBDV strains MD, GLS, and IN and the classic IBDV strains SAL and STC but did not detect antibodies against the variant Del-A and classic IBDV strain BVM or the serotype 2 IBDV strain OH. PMID- 9356709 TI - Restriction fragment length polymorphisms in the VP2 gene of infectious bursal disease viruses. AB - Infectious bursal disease viruses (IBDVs) were examined for restriction fragment length polymorphisms in a fraction of the VP2 gene with the use of the reverse transcriptase/polymerase chain reaction-restriction fragment length polymorphism (RT/PCR-RFLP) assay. The restriction enzymes BstNI and Mbol were used to obtain RFLP results. A third enzyme, StyI, was tested, but its utility for differentiation of IBDV strains was limited. Thirteen vaccine viruses and five IBDV strains that were previously characterized were placed into five molecular groups. Two groups contained viruses described as being classic strains, and two groups contained viruses described as being variant strains. The fifth group contained both classic and variant strains. The RFLP observed for the serotype 2 IBDV strain OH was unlike any of the RFLPs observed in viruses in the five molecular groups. Seven IBDV strains from commercially reared chickens in the United States, Mexico, Puerto Rico, and Thailand were tested in the RT/PCR-RFLP assay to determine if they were similar to the commercial IBDV vaccine strains tested. These viruses were selected because they were associated with lesions in the bursa of chickens that should have been protected by maternal antibodies or active immunity. Each of the viruses tested contained a unique RFLP compared with the IBDV strains and vaccine viruses examined in this study and, thus, did not fit into any of the five molecular groups. These viruses also were distinguishable from each other. PMID- 9356710 TI - Appearance of T cells in the bursa of Fabricius and cecal tonsils during the acute phase of infectious bursal disease virus infection in chickens. AB - Groups of 3-wk-old specific-pathogen-free chickens were inoculated intraocularly with two virulent and one vaccine strain of infectious bursal disease virus (IBDV). During the acute phase of infection, the bursa of Fabricius and cecal tonsil were examined by immunohistochemistry for IBDV antigen and T cell. With all three viruses examined, the infection resulted in the appearance of viral antigen in the bursa accompanied by the presence of CD3+ cells. The CD3+ cells were predominantly TCR2+ and appeared at the site where viral antigens were present. The CD3+ cells continued to persist in the bursa after most of the IgM+ cells and IBDV antigen-positive cells had disappeared. Similar appearance of CD3+ cells was noted following viral antigen accumulation in the germinal centers of cecal tonsils in virulent IBDV-infected chickens. These results demonstrated that infection with IBDV resulted in extensive viral replication in the bursa and the cecal tonsils with an associated accumulation of T cells. The role these T cells may play in the pathogenesis of IBDV infection remains to be established. PMID- 9356711 TI - Polymerase chain reaction to detect infectious laryngotracheitis virus in conjunctival swabs from experimentally infected chickens. AB - The polymerase chain reaction (PCR) was standardized and assessed as a potential diagnostic test for infectious laryngotracheitis using conjunctival swabs collected from experimentally infected chickens. Polymerase chain reaction primers based on the sequence of a 1.1-kb BamHI restriction enzyme fragment of the Ontario 1598 (Ont 1598) strain of infectious laryngotracheitis virus were selected and 300 fg of purified viral DNA were detected by ethidium bromide staining of agarose gels or 30 fg were detected by DNA hybridization. At least five different strains (Ont 1598, ATCC N-71851, LT-IVAX, Laryngo-Vac, and a local strain 322) were amplified whereas other avian pathogens and uninfected cell cultures tested negative. Swabs collected from experimentally infected chickens were analyzed by both PCR and virus isolation on various days postinfection. A comparison of virus isolation to PCR indicated that, in the mid-postinfection phase, PCR and virus isolation appeared to be comparable with a kappa value of greater than 0.8. The polymerase chain reaction was shown to be the better test later in infection, when clinical recovery had occurred. PMID- 9356712 TI - Amplification of avian reovirus RNA using the reverse transcriptase-polymerase chain reaction. AB - A reverse transcriptase-polymerase chain reaction method was developed for the detection of avian reovirus. The origin of primers was from the S1 gene of the avian reovirus genome. A reovirus-specific 532-base pair cDNA product was amplified by these primers from six reference strains and 23 field isolates of avian reoviruses, but not from seven different avian pathogenic viruses and bacteria. As little as 1 pg of avian reovirus RNA was detected using gel electrophoresis and Southern blot hybridization. PMID- 9356713 TI - Antigenic and S-1 genomic characterization of the Delaware variant serotype of infectious bronchitis virus. AB - A previously unrecognized infectious bronchitis virus (IBV) serotype, referred to hereafter as the Delaware variant (DE var), was isolated from commercial broiler chickens during a severe, widespread respiratory disease epornitic in the Delmarva peninsula region of the United States in January-March 1992. The DE var serotype was found to be antigenically unrelated by virus-neutralization (VN) test to nine reference IBV serotypes from North America. Additional VN tests indicated that the DE var isolates (DE/072/92, DE/121/ 92, DE/152/92, and DE/174/92) from broilers were fully or partially neutralized by monospecific antisera prepared against themselves and against two IBV field isolates (DE/492/90 and DE/903/90) recovered from a Delmarva commercial layer flock experiencing egg production losses in 1990. Antigenic relatedness values determined by VN indicated layer isolate DE/492/90 was more closely related to the broiler DE var isolates than was layer isolate DE/903/90. Cross-challenge tests performed in specific-pathogen-free chickens also demonstrated the antigenic similarity of the broiler (DE/072/92 and DE/174/92) and the layer isolates (DE/492/90 and DE/903/90), with heterologous strain protection values ranging from 55% to 100%. Protection values of DE var isolates vs. Massachusetts 41 and Arkansas DPI were considerably lower (0-60%). The S-1 gene of the US/DE/072/92 isolate of the DE var serotype was amplified by reverse transcription polymerase chain reaction, cloned, and sequenced. The DE var S-1 gene sequence was compared with the S-1 gene sequences of IBV serotypes from North America, Europe, and Australia. A dendrogram based on this analysis supported the conclusion that the DE var serotype is highly novel among IBV. A high degree of similarity (> 88%) was observed between the S-1 genes of the DE var broiler isolates (DE/072/92 and DE/174/92) and layer isolates (DE/492/90 and DE/903/90). These data, taken with the VN and cross-challenge results, establish a genetic as well as an antigenic link between the isolates from layers and broilers and indicate the DE var serotype was responsible for both infectious bronchitis outbreaks. PMID- 9356714 TI - Pathogenicity of a strain of Trichomonas gallinarum in turkeys and its possible interaction with cecal coccidia. AB - The pathogenicity of Trichomonas gallinarum (TG) in turkeys and chickens was assessed in a series of four experiments. TG was shown to be resistant to freezing for a period of 1 hr at -20 C; birds administered an emulsion of previously frozen TG were readily infected. Young birds receiving this inoculum were more likely to be infected with TG in both ceca compared with birds administered TG emulsion that had remained at room temperature for the same length of time. In this and other experiments, birds infected with the parasite consistently produced a yellow frothy liquid in the ceca, as well as small raised papulae on the mucosal surface of the ceca. Histologically, the lesions were located in the lamina propria, with openings that extended from the apex of the lesion to the crypts. The lamina propria was consistently infiltrated by lymphocytes and scattered heterophiles. Although TG is likely involved in the pathogenesis of the lesions, the resulting pathology could not be linked definitively to TG alone because inoculation was performed with a cecal contents homogenate containing significant numbers of cecal bacteria. Combined infections of TG and Eimeria adenoides (EA) were also studied. Turkeys administered both parasites were more frequently infected with TG in both ceca compared with those that received TG alone. Ceca infected with TG alone tended to be enlarged and gas filled, whereas those infected with the combination of TG and EA were smaller and usually lacked the yellow frothy liquid contents. PMID- 9356715 TI - Detection of Pasteurella multocida-specific DNA in turkey flocks by use of the polymerase chain reaction. AB - A polymerase chain reaction (PCR)-based assay using primers constructed to amplify the gene (psl) encoding the P6-like protein (Psl) of Pasteurella multocida was developed. After Southern blotting and hybridization with psl, the assay (PCR-H) was found to be specific (it did not detect a variety of other avian bacterial pathogens) and sensitive (detected > or = 10 P. multocida organisms or > or = 24 femtograms of extracted P. multocida DNA). Samples were collected from the oropharynx of randomly selected birds housed on premises that had recently experienced an outbreak of avian cholera (outbreak farms) or from birds housed on premises that had not reported an outbreak of this disease during the preceding 12 mo (control farms). The PCR-H assay detected 11 infected turkeys out of a total of 178 sampled on six outbreak farms as compared with isolation of P. multocida from 23 turkeys by using mouse inoculation. Neither method detected P. multocida in samples collected from 174 turkeys sampled on six control farms. Statistical analysis using the Kappa test demonstrated that the results of the two tests showed poor agreement from five outbreak flocks (K = 0, 0, 0, 0.35, 0.47) and strong agreement from one outbreak flock (K = 0.89). Combined results from the outbreak flocks showed poor agreement (K = 0.49) between the two methods. PMID- 9356716 TI - Biological and molecular characterization of Newcastle disease virus isolates from surveillance of live bird markets in the northeastern United States. AB - Newcastle disease virus (NDV) is frequently recovered from surveillance samples collected by U.S. Department of Agriculture, Animal and Plant Health Inspection Service personnel in live bird markets. Six NDV isolates, five from chickens and one from a pheasant, were characterized for comparison with reference NDV isolates from poultry and other birds. All isolates tested were of low virulence for chickens. Four of the six isolates were similar to reference lentogens B1 and La Sota, but two isolates, one from a chicken and one from a pheasant, were different. The aberrant chicken isolate had a monoclonal antibody-binding profile like an unusual Canadian pigeon isolate. Sequence analysis of the matrix gene of this isolate demonstrated that it differed from all isolates included in the comparison and therefore may represent a third phylogenetic NDV group. The pheasant isolate had a monoclonal antibody-binding profile typical of other U.S. NDV lentogens but had a matrix gene sequence and hemagglutinin thermostability similar to strains Ulster and Queensland V4 (QV4), viruses originally isolated in Northern Ireland and Australia, respectively. The pheasant virus is the first lentogen isolated in the United States known to be closely related phylogenetically to Ulster and QV4. The unusual chicken and pheasant isolates were readily shed from the intestinal tract during chicken passage, whereas the other isolates were shed from the respiratory tract with little or no intestinal shedding. The frequency in live bird markets of viruses similar to those previously thought to be exotic to the United States in unknown. PMID- 9356717 TI - Cytotoxic activity of cells recovered from the respiratory tracts of chickens inoculated with infectious bronchitis virus. AB - Previously uninoculated control chickens and chickens exposed to infectious bursal disease virus (IBDV) at 1 day of age were intranasally exposed to the M41 strain of infectious bronchitis virus (IBV) at 5 wk of age. Between 7 and 13 days after inoculation with IBV, cells were collected from the respiratory tracts of both groups of chickens and assayed for in vitro cytotoxic activity against a lymphoblastoid LSCC-RP9 target cell line using a 4-hr 51chromium-release assay (CRA). Compared to thymocytes from uninfected chickens, which were used as negative controls in the CRA, respiratory tract cells from both groups consistently displayed significant cytotoxic activity against LSCC-RP9 target cells. This cytotoxic activity, attributed to natural killer cells, was statistically more pronounced (P < 0.05) in IBDV plus IBV-infected chickens than in chickens inoculated with IBV alone. PMID- 9356718 TI - Comparative evaluation of in vitro and in vivo assays for the detection of reticuloendotheliosis virus as a contaminant in a live virus vaccine of poultry. AB - Indirect immunofluorescence (IFA), polymerase chain reaction (PCR), and enzyme linked immunosorbent assay (ELISA) were used for detection of reticuloendotheliosis virus (REV) as a contaminant in a live virus fowl pox (FP) vaccine of poultry. A FP vaccine known to be contaminated with REV was tested by in vitro and in vivo assays in chicken embryo fibroblasts (CEFs) and day-old specific-pathogen-free (SPF) chicks, respectively. Using in vitro assays, IFA and PCR were more sensitive than ELISA in detection of REV in CEFs inoculated with REV-contaminated FP vaccine. However, when the vaccine was tested by in vivo assays using SPF chickens, the sensitivity of ELISA was comparable with that of IFA and PCR. Antibody to REV was not detected in SPF chickens within 4 wk postinoculation with REV-contaminated FP vaccine at hatch. Filtration of vaccine to eliminate vaccine virus from the inoculum before testing in CEFs resulted in a significant reduction in the frequency of REV detection by PCR or IFA. The data suggest that the sensitivity of IFA, PCR, and ELISA depends on the concentration of REV in the vaccine and that in vivo assays of vaccines for contamination with REV should include a test for virus because a negative antibody test may be misleading. PMID- 9356719 TI - Protection against coccidiosis in outbred chickens elicited by gamma-irradiated Eimeria maxima. AB - In an effort to develop an attenuated coccidiosis vaccine against coccidiosis, we exposed Eimeria maxima oocysts to an optimum dose of gamma irradiation (17 kRad) that does not affect sporozoite invasion of the intestinal mucosa but does prevent asexual parasite development. Irradiated E. maxima oocysts were suspended in gelatin slabs and placed in battery cages for ingestion by 1-day-old chickens. Separate groups of chickens were given gelatin slabs containing nonirradiated E. maxima oocysts or were inoculated per os with either irradiated or nonirradiated E. maxima oocysts. Chickens infected with irradiated or nonirradiated oocysts by either oral inoculation or gel delivery showed a dose-dependent protection against weight loss associated with E. maxima challenge compared with unimmunized controls. In general, nonirradiated oocysts elicited protective immunity at lower immunization doses compared with irradiated oocysts. These experiments were extended to a floor pen study wherein 1-day-old male and female broiler chickens were given irradiated or nonirradiated E. maxima oocysts in gelatin slabs in hatching boxes and challenged at 4 wk of age. A significant reduction (P < 0.05) in lesion scores was observed for chickens immunized with either irradiated or nonirradiated oocysts compared with unimmunized controls. Although no significant difference (P > 0.05) was observed in weight gain between these groups, both male and female chickens inoculated with irradiated E. maxima oocysts showed about a 10% greater weight gain than unimmunized controls. For both male and female chickens, average weights at challenge were greater in groups that were immunized with 17-kRad-irradiated E. maxima oocysts compared with those animals immunized with nonirradiated oocysts. PMID- 9356720 TI - Comparison of drag-swab environmental protocols for the isolation of Salmonella in poultry houses. AB - Three surveys were conducted during November 1995 and March and May 1996 to compare the use of double-strength skim milk (wet) or no transport media (dry) drag swabs for the detection of salmonellae in 10 broiler houses. Salmonellae were isolated from 57 of 120 individual wet drag-swab samples, compared with 21 of 120 dry drag-swab samples. Furthermore, Salmonella was detected at a higher frequency with wet drag swabs (66.7%) than with dry drag swabs (40%) when compared on an individual growout house basis. A total of seven different serotypes were isolated from the 10 broiler houses. Although double-strength skim milk drag swabs are more labor intensive than dry drag swabs, double-strength skim milk drag swabs are more efficient for detecting Salmonella than are dry drag swabs with no transport media. PMID- 9356721 TI - Analysis of avian lymphocyte proliferation by a new, simple, nonradioactive assay (lympho-pro). AB - An assessment of T-cell-mediated immune functions (i.e., lymphocyte proliferation assay) in the chicken, unlike the determination of antibody levels, is not routinely performed. This is primarily because of difficulties in the isolation of relatively pure populations of lymphocytes and the use of radioactive isotopes. To address these issues, the goals of our study were to optimize a method for isolating and enriching avian lymphocyte populations and to develop a nonradioactive lymphocyte proliferation assay. To accomplish these goals, we used a multiple slow-speed centrifugation technique combined with a "swirl" collection technique for lymphocyte isolation from chicken peripheral blood. After a fraction enriched with lymphocytes was obtained, a simple, rapid colorimetric and fluorometric assay (lympho-pro) to indirectly determine mitogen-induced proliferation was adapted and compared with the "Gold Standard" [3H]thymidine. Chickens of different ages and two genetic strains were used in this study. Lymphocytes were stimulated with various concentrations of concanavalin A (Con A, T-cell mitogen) or phorbol 12-myristate 13-acetate + ionomycin (pan lymphocyte mitogen). Our studies showed that the pattern of lymphocyte proliferation assessed by the Alamar blue-based lympho-pro assay was similar to the [3H]thymidine incorporation assay. Younger birds had higher levels of mitogen induced proliferation when compared with adults of the same genetic strain. Because the lympho-pro assay, unlike [3H]thymidine, does not require lysis of cells to assess proliferation, cells that have undergone stimulation/proliferation can be subsequently characterized by staining with antibodies against cell surface antigens and analysis by flow cytometry. Another notable advantage of the lympho-pro assay is the rapidity of assessment and nontoxicity. In conclusion, this assay may be of value in assessing some aspects of T-cell-mediated immunity in both avian research and avian medicine diagnostic settings. PMID- 9356722 TI - Characterization of an avian adenovirus associated with inclusion body hepatitis in day-old turkeys. AB - A group I avian adenovirus isolated from day-old turkeys with inclusion body hepatitis (IBH) was identified as turkey adenovirus serotype 2 (TAV2) based on cross-neutralization assays and DNA restriction endonuclease analyses. Yolk sac inoculation of embryonated turkey eggs resulted in embryo mortality and significantly (P < 0.01) decreased hatchability compared with sham-inoculated controls. Embryo mortality occurred primarily between day 24 of incubation and the time embryos hatched. Focal necrosis was detected in livers of 11/52 virus inoculated embryos that died postinoculation and 1/27 hatchlings; in three embryos, areas of necrosis contained intranuclear inclusion bodies. These findings identify the IBH isolate as TAV2, incriminate the virus as a potential cause of suboptimal hatchability in turkeys, and provide additional evidence for causal involvement in IBH. PMID- 9356724 TI - Severe hepatitis resulting from toxoplasmosis in a barred owl (Strix varia) from Quebec, Canada. AB - A female adult barred owl (Strix varia) had been hurt by a car. Its general status declined gradually within 2 wk with anorexia and inactivity. Necropsy examination revealed marked multifocal pale areas in liver, emaciation, and mild airsacculitis and pericarditis. Histopathologic examination revealed severe acute multifocal hepatic necrosis with numerous protozoal tachyzoites within necrotic foci and in the cytoplasm of hepatocytes and macrophages. These tachyzoites stained with an indirect immunohistochemistry method for Toxoplasma gondii antigens. This is the first reported case of hepatitis resulting from toxoplasmosis in a raptor. PMID- 9356723 TI - Genotypic evaluation of Salmonella enteritidis isolates of known phage types by arbitrarily primed polymerase chain reaction. AB - Salmonella enteritidis isolates of known phage types 8, 13a, and 14b were inoculated separately into a group of 14-wk-old specific-pathogen-free chickens. S. enteritidis isolates from pre- and postinoculum were analyzed with the use of arbitrarily primed polymerase chain reaction to determine any genotypic changes after reisolation from the chickens. The preinoculum isolates and the isolates of S. enteritidis recovered at 5, 10, 20, and 35 days postinoculation from the inoculated chickens with various phage types were similar. Therefore, no changes of S. enteritidis organisms of similar phage types occurred at the genotypic level. However, SE-3-B7001 (phage type 8) was changed to phage type 14b after inoculation into the chickens. PMID- 9356725 TI - Septicemia associated with Hafnia alvei in laying hens. AB - In the present work Hafnia alvei was isolated from laying hens displaying a reduction in egg production, loss of appetite, opisthotonus, and death. Multifocal necrotizing hepatitis and splenitis were the most prominent lesions. The organism was identified microbiologically. Laying hens were experimentally inoculated by the oral and intraperitoneal route to show the pathogenicity of the organism. A very similar clinicopathologic effect resulted from this trial. Several experimentally infected laying hens died due to septicemia. We conclude that H. alvei may cause a septicemia similar to that reportedly caused by Salmonella spp. in avian species. PMID- 9356726 TI - Intestinal cryptosporidiosis in pigeons (Columba livia). AB - An intestinal disease in pigeons (Columba livia) from the Canary Islands characterized by diarrhea and body weight loss is described. Intestinal cryptosporidiosis was identified in three young pigeons. Cryptosporidia were associated with hyperplasia of the intestinal crypts and moderate inflammatory infiltration in lamina propria. This is the first report of cryptosporidiosis in pigeons. PMID- 9356727 TI - Metastatic pheochromocytoma in a parakeet. AB - Pheochromocytoma, a tumor of the adrenal medulla, was diagnosed in a budgerigar based on gross, histopathologic, and electron microscopic findings. PMID- 9356728 TI - Basal cell carcinoma in a blue-fronted amazon parrot (Amazona aestiva). AB - Tumors of the integumentary system are relatively common in companion birds. Dermal tumors in pet birds can be epithelial, mesenchymal, or vascular in origin. Basal cell carcinomas appear to be extremely rare in birds. An adult female blue fronted Amazon parrot was examined because it exhibited bilateral cervical masses that extended from the base of the skull to the ingluvial region. The tumors were removed by surgical excision. Microscopic examination of the masses revealed neoplastic epithelial cells that extended to all borders of the sections; scattered vessels with neoplastic cells within their lumens were also found. The histopathologic diagnosis was basal cell carcinoma. Six weeks postoperatively, the masses recurred and the bird was euthanatized. This report suggests that basal cell carcinomas should be considered as a differential for avian dermal tumors. This neoplastic condition can be aggressive and has the potential to metastasize. PMID- 9356729 TI - Bone mineral density reflects bone mass but also the degree of mineralization of bone: therapeutic implications. PMID- 9356730 TI - Estrogen receptor gene polymorphism and bone mineral density at the lumbar spine of pre- and postmenopausal women. AB - In order to analyze the role of the estrogen receptor (ER) gene allelic polymorphisms on bone mineral density (BMD), 173 pre- and postmenopausal women were divided into four groups according to their menstrual status (group A: premenopausal women; group B: late premenopausal women; group C: postmenopausal women who had menopause for 5 years or less; and group D: postmenopausal women who had menopause for more than 5 years), and the relationship between ER gene polymorphism and lumbar spine BMD, the percent annual change in BMD and biochemical markers were studied. The restriction fragment length polymorphism (RFLPs) were represented as Xx (XbaI) and Pp (PvuII), with upper case and lower case letters signifying the absence or presence of restriction sites, respectively. In group A, the Xx genotype had significantly higher BMD (p < 0.01) than the xx genotype, but the difference was lost in groups B, C, and D. Because the percent annual change in BMD of group A was 0.052% and was not statistically different among genotypes, it is suggested that RFLP by Xba I is closely linked with peak bone mass that was attained during the subject's late thirties. In group B, serum N-region osteocalcin (N-OC) levels and the percent annual change in BMD showed a significantly larger increase than that of group A, indicating postmenopausal bone loss had commenced. Because the N-OC level of the Xx genotype was significantly higher than that of the xx genotype (p < 0.05), and the percent annual change in BMD of the Xx genotype showed a tendency to increase (p = 0.072), it is suggested that the high BMD of the Xx genotype is rapidly lost during menopausal transition. There were no significant relationships between RFLP and BMD in groups C and D, and between RFLP and BMD in groups C and D, and between RFLP by PvuII and BMD. The present study suggests that the Xx genotype is involved in accretion of BMD during young adulthood, but the effect was lost during menstrual transition. PMID- 9356731 TI - Temporal expression of PTHrP during endochondral bone formation in mouse and intramembranous bone formation in an in vivo rabbit model. AB - Expression of parathyroid hormone-related protein (PTHrP) messenger RNA (mRNA) and protein was investigated throughout the developmental progression of endochondral bone formation in mouse and intramembranous bone formation in an in vivo model in rabbit, using in situ hybridization and immunohistochemistry. Endochondral bone formation was investigated in a developing embryo, newborn, and adult mouse. In fetal long bones through to newborn (day 7), PTHrP mRNA and protein were consistently expressed in chondrocytes within the proliferative, transitional, and hypertrophic zones. In addition, high levels of PTHrP were also detected in osteoblasts on the surface of trabecular bone surfaces. Similarly, at the adult stage (week 7), PTHrP mRNA and protein were consistently expressed in chondrocytes at epiphyseal ends of the subarticular cartilage, within cortical periosteum, as well as in osteoblasts located at the metaphyseal trabecular bone surfaces. Using an in vivo intramembranous bone formation model in rabbits, expression of PTHrP mRNA and protein was demonstrated in preosteoblasts prior to trabecular bone formation (1-week bone harvest). As bone formed (2-, 3-, and 4 week bone tissue harvests), PTHrP mRNA and protein were highly expressed in actively synthesizing osteoblasts and in those osteocytes embedded within the superficial layers of the bone matrix. Lining osteoblasts and osteocytes buried deeply in the bone matrix displayed weak or no signal for PTHrP. The pattern of spatial and temporal expression of PTHrP demonstrated in cartilage cells and osteoblasts in the two systems suggests an important role of PTHrP in both endochondral and intramembranous bone formation. PMID- 9356732 TI - Purinergic transmitters inhibit bone formation by cultured osteoblasts. AB - Adenosine triphosphate (ATP) and other purinoceptor agonists cause a transient rise in [Ca2+]i in cultured osteoblast-like cells and have a mitogenic effect, as does parathyroid hormone (PTH), and there is evidence that ATP and PTH can act synergistically on osteoblasts. The likelihood that nucleotides, acting through purinoceptors, are important local factors in bone remodeling is therefore considerable. However, their effect on bone formation is unknown. We recently developed a culture system in which appositional bone formation occurs only in narrow grooves cut in a substratum. We have used this as an assay to measure the effects of ATP (50 and 500 mumol/L), ATP gamma S (20 mumol/L), 2-MeSATP (2 and 20 mumol/L), uridine triphosphate (UTP) (0.2, 2, and 20 mumol/L), adenosine (20 mumol/L), bovine PTH (0.25 and 0.5 IU/mL), rat PTH1-34 (10(-8) and 10(-7) mol/L), and rat PTHrP1-40 (10(-9) and 10(-8) mol/L) on bone formation by rat calvarial osteoblasts. The culture medium was renewed 3 times/week (every 2 or 3 days), and the number of bone loci and length and area of Alizarin red-stained mineralized bone formed in the grooves of each specimen in 16-29 days were measured. Compared with controls, ATP gamma S, 2-MeSATP, and ATP reduced the amount of bone formed in a 2-3 week culture period. Adenosine had no effect, and UTP either had no effect or at 2 mumol/L stimulated bone formation. PTH and PTHrP completely abolished bone formation in 4 week cultures. Our findings are consistent with evidence for more than one P2 purinoceptor subtype in bone, and show for the first time that the effect of ATP on appositional bone formation by osteoblasts in vitro is, like PTH and PTHrP, inhibitory. PMID- 9356733 TI - Application of automatic image segmentation to tibiae and vertebrae from ovariectomized rats. AB - Automatic contextual segmentation algorithms were developed to objectively identify bone compartments in pQCT images of tibiae, femora, and vertebrae. Principal advantages of this approach over existing techniques such as histomorphometry are as follows: (a) the algorithms can be implemented in a fast, uniform, nonsubjective manner across many images, allowing unbiased comparisons of therapeutic efficacy; (b) much larger volumes in the region of interest can be analyzed to derive true volumetric parameters for trabecular and cortical bone compartments; and (c) pQCT can be used to quantitate bone effects longitudinally in vivo. An automatic contextual segmentation algorithm was used to analyze over 600 scans of proximal tibiae, distal femora, and L-4 vertebrae from studies with ovariectomized rats. Accuracy and precision analyses were performed, and correlation to histomorphometry parameters showed that pQCT trabecular bone density correlates to Tb.N with r = 0.93, while BV/TV correlates to Tb.N with r = 0.95. In other words, pQCT correlates as well to histomorphometry as histomorphometry does to itself. We conclude that the developed automatic segmentation algorithm provides fast, precise, and objective quantitation of bone compartments that are highly correlated with histomorphometry measurements. PMID- 9356734 TI - Calcium hydroxide ameliorates tobramycin toxicity in cultured chick tibiae. AB - Results from this laboratory have shown that bone metabolism is directly related to extracellular pH and that high concentrations of tobramycin released from impregnated polymethylmethacryrate (PMMA) beads has pH-dependent toxic effects on bone. In the present study, beneficial effects of calcium hydroxide-impregnated PMMA were investigated regarding tobramycin toxicity and bone metabolism in chick embryo tibiae in vitro. Also using Ca(OH)2 as a pH regulator, the antibiotic efficacy of tobramycin-impregnated PMMA was evaluated with respect to inhibition of Staphylococcus aureus growth. When Ca(OH)2 was added to PMMA beads containing tobramycin, the beads released hydroxyl and calcium ions into the culture medium and released more antibiotic than beads containing only tobramycin. Bone metabolism (glycolysis, total protein synthesis, and collagen synthesis) was enhanced by Ca(OH)2-impregnated beads with or without tobramycin. Additionally, bacterial growth was inhibited more strongly when S. aureus was incubated with tobramycin- and Ca(OH)2-impregnated PMMA disks than with disks containing only tobramycin. This study demonstrates the feasibility of adding Ca(OH)2 to tobramycin-impregnated PMMA beads as a regulator of local pH and a promoter of bone metabolism for protection of bone when high concentrations of tobramycin are used to treat osteomyelitis. It also suggests that lower concentrations of antibiotic may be effective if Ca(OH)2 and tobramycin are administered simultaneously. PMID- 9356735 TI - Changes in canine cortical and cancellous bone mechanical properties following immobilization and remobilization with exercise. AB - The purpose of this study was to assess cortical and cancellous bone responses to unilateral limb immobilization and, subsequently, to remobilization with exercise, in a young adult canine model. Right forelimbs of 14 1-2-year old mongrel dogs were immobilized in a non-weight-bearing position by a bandage for 16 weeks. Six control dogs were untreated. At 16 weeks, seven immobilized and three control dogs were euthanized. The remaining seven immobilized dogs began a recovery protocol consisting of 16 weeks of kennel confinement (without the right forelimb bandaged) followed by 16 weeks of treadmill exercise conducted three times per week. These seven dogs and three control dogs were euthanized at 48 weeks. Bone mineral density of the proximal radii was determined with dual-energy X-ray absorptiometry and humeral middiaphyseal cross-sectional areas were determined with computed tomography. Humeri were tested in cranio-caudal three point bending to failure. Cancellous bone cores from the lateral humeral condyles had wet apparent density determined and were tested to failure in compression. Mechanical properties, bone density, and cross-sectional areas were compared between immobilized (right forelimb), contralateral weight bearing (left forelimb), and control forelimbs with Kruskal-Wallis and post hoc tests. At 16 weeks, bone mineral density, cortical load, yield, and stiffness as well as cancellous bone failure stress, yield stress, and modulus were significantly lower (p < 0.02) for immobilized limbs than control limbs. Immobilized limb cancellous bone mechanical properties were 28%-74% of control values, and cortical bone mechanical properties were 71%-98% of control values. After 32 weeks of remobilization, cortical and cancellous bone mechanical properties were not different from control values except that cortical bone failure stress and modulus were significantly higher (p < 0.01) between remobilized and control limbs. In summary, 16 weeks of forelimb immobilization was associated with significantly lower mechanical properties, and with greater differences in cancellous than cortical bone properties. Mechanical properties were not different from control values after 32 weeks of recovery that included 16 weeks of treadmill exercise. PMID- 9356736 TI - Effect of enzyme replacement therapy on bone formation in a feline model of mucopolysaccharidosis type VI. AB - A range of skeletal abnormalities are evident in mucopolysaccharidosis type VI (MPS VI, Maroteaux-Lamy syndrome) including short stature and dysostosis multiplex, resulting from a deficiency in the lysosomal hydrolase N acetylgalactosamine-4-sulphatase (4S). In this article, bone pathology was assessed in a feline model of MPS VI to evaluate the efficacy of enzyme replacement therapy (ERT) as a treatment modality for this genetic disorder. Osteopenia is clearly evident in MPS VI animals, with bone mineral volume (BV/TV) falling well below that of normal animals (4.39% vs. 20.11%, respectively). Trabecular bone architecture was also affected in MPS VI with fewer, thinner, and more widely spaced trabeculae apparent. Bone formation rate (BFR/BS) was also lower in MPS VI animals than controls (0.0011 mm3/mm2 per day vs. 0.008 mm3/mm2 per day, respectively). Vertebral and tibial bone length in MPS VI animals progressively fell behind normal values with increasing age, as did cortical bone thickness. Vertebral body shape was also altered. ERT with recombinant human 4S (rh4S) resulted in a vertebral BV/TV of 8.23% in animals treated with an intravenous enzyme dose of 1 mg/kg and a BV/TV of 14.33% in animals treated with a dose of 5 mg/kg. BFR/BS also increased to 0.0034 mm3/mm2 per day in animals treated with enzyme doses of either 1.0 or 5.0 mg/kg rh4S. All other affected histomorphometric parameters also improved with ERT to a level intermediate between MPS VI untreated animals and normals. However, individual animals treated with 0.2 mg/kg rh4S intravenously or 1.0 mg/kg rh4S administered subcutaneously did not exhibit an improvement over untreated MPS VI animals. Vertebral and tibial bone lengths, tibial cortical bone thickness, and vertebral body shape also responded to ERT, with a trend away from the untreated group. Thus, ERT had a positive effect on bone development in MPS VI animals that was dependent upon the dose of enzyme administered and the route of administration. PMID- 9356737 TI - Aromatase cytochrome P450 transcripts are detected in fractured human bone but not in normal skeletal tissue. AB - Peripheral conversion of gender steroid precursors has been implicated in playing a role in bone turnover in postmenopausal women. It has been reported that aromatase cytochrome P450 (P450arom) is present in primary bone and bone marrow (BM), and that P450arom mRNA has been identified in cultured BM and osteoblast like cell lines. However, there are no reports that P450arom transcripts have been detected in skeletal tissue that has not been cultured. We therefore elected to test for the presence of P450arom mRNA in primary human bone and BM in normal and fractured necks of femora using the reverse transcription-polymerase chain reaction method. Although the RNA extracted from these tissues was of good quality as demonstrated by the expression of transcripts for interleukin-6, P450arom transcripts failed to be detected in normal primary cortical bone and fatty BM containing trabecular bone. However, P450arom transcripts were detected in the latter when they were cultured. Transcripts for P450arom were also detected in total RNA extracted from six fractured necks of femora and semiquantitative PCR demonstrated that P450arom mRNA was present in similar abundance in the same amount of RNA analyzed from buttock adipose tissue and fractured bone/BM. P450arom mRNA expression was also detected in cultured peripheral blood leukocytes, suggesting that this might be the source of the enzyme. In these cultures no correlation was detected between the expression of P450arom mRNA and cell proliferation. PCR failure was excluded in cases when P450arom transcripts failed to be detected in bone/BM by coamplifying RNA from human and rat brain mRNA, known to express P450arom mRNA, using primers that detect both P450arom mRNA from both species. These products were analyzed by Southern blot using oligonucleotide probes, which label either human or rat P450arom cDNA. The blots confirmed the absence of P450arom in nonfractured human bone and BM and preclude PCR failure. Our results indicate that P450arom mRNA is not detected in either normal human bone or BM, but can be induced in this microenvironment under pathological conditions. We propose that tissue grown in vitro is analogous to a wound and this explains why P450arom transcripts were detected in cultured normal skeletal tissue, whereas they failed to be detected in primary normal bone and BM. PMID- 9356738 TI - Mandibular condyle bone mineral density measurement by quantitative computed tomography: a gender-related difference in correlation to spinal bone mineral density. AB - We conducted volunteer studies to assess age-related changes of mandibular condyle bone mineral density (BMD) and its correlation to the spinal BMD. Quantitative computed tomography was performed on the condyles and spines (L1-3) of 210 healthy subjects (114 men and 96 women, aged 5-85 years). A separate study was performed on 73 young student subjects (39 men and 34 women, aged 23-25 years). The mandibular condyle BMD showed a decrement rate similar to spinal BMD in men, but in women the decrement rate of the mandibular condyle BMD was lower than that of the L1-3 BMD. On the other hand, correlation coefficients in BMD between the mandibular condyle and spine were similar in women and men. Gender related differences were found to be dramatic when assessed in the young student group; the mandibular condyle and spinal BMDs were highly correlated in women (r = 0.82, p < 0.0001), but no correlation was found in men (r = 0.22). Taken together, these results suggest that the same regulatory mechanisms exist in the mandibular condyle and spine BMDs. However, aside from the spine BMD, additional undefined factor(s), including mechanical stress from the occlusion, may be involved in maintaining mandibular BMD. PMID- 9356739 TI - The benefit of hormone replacement therapy on bone mass is greater at the vertebral body than posterior processes or proximal femur. AB - The aim of this study was to determine whether the higher vertebral bone mass in women receiving hormone replacement therapy (HRT) is confined to the trabecular rich vertebral body rather than the predominantly cortical posterior processes, and to determine whether the protective effect of HRT at the proximal femur, a predominantly cortical site, is less than at the spine. Bone mass (g) of the third lumbar vertebra (total, vertebral body and posterior processes, measured by lateral scanning), and bone mineral density (g/cm2) of the femoral neck, Ward's triangle, and trochanter were measured using dual X-ray absorptiometry in a cross sectional study of 71 women receiving HRT for 5.7 +/- 0.4 years (mean +/- SEM), ranging from 1 to 21 years, 69 age-matched controls, and 42 premenopausal controls aged 20 to 40 years. Relative to untreated postmenopausal controls, total bone mass of the third lumbar vertebra (body plus posterior processes) by postero-anterior (PA) scanning was 0.4 +/- 0.1 SD or 9.6 +/- 3.0% higher in HRT treated women (p < 0.01). By lateral scanning, total bone mass was higher than age-matched controls (z score 0.4 +/- 0.1 SD or 11.2 +/- 3.4%, p < 0.01). This difference was confined to the vertebral body (z score 0.6 +/- 0.1 SD, p < 0.001), which was 17.1 +/- 3.3% higher than in age-matched controls (p < 0.001). Bone mass of the posterior processes was no higher [z score 0.1 +/- 0.1, not significant (NS)]. The deficit at the vertebral body in HRT-treated women, relative to premenopausal controls, was half the deficit at the vertebral body in untreated postmenopausal women (t score -0.7 +/- 0.1 vs. -1.4 +/- 0.1 SD, respectively; p < 0.001) but no less at the posterior processes (t score -1.6 +/- 0.2 vs. -1.9 +/- 0.2 SD, respectively; NS). Similarly, the deficit in the vertebral body in the HRT treated group was half the deficit at their posterior processes (t score -0.7 +/- 0.1 SD vs. -1.6 +/- 0.2, respectively; p < 0.001). In HRT-treated women, bone mass diminished significantly with age at the posterior processes (r = -0.31, p < 0.01), but not at the vertebral body (r = -0.21, p = 0.07). Bone mass diminished significantly with age at the vertebral body and posterior processes in untreated women (r = -0.55, p < 0.001; r = -0.45, p < 0.001, respectively). Bone density (g/cm2) diminished at all femoral sites with advancing age in HRT-treated women. A protective effect was seen at the femoral neck and Ward's triangle, but not trochanter (z score 0.2 +/- 0.1, p = 0.06; 0.3 +/- 0.1, p < 0.05; 0.0 +/- 0.1, NS, respectively). In conclusion, the protective effect of HRT against bone loss at the vertebral body, the site of fracture in osteoporosis, may be underestimated by PA scanning. The greater benefit at the vertebral body, and more modest effect at the proximal femur, suggests that HRT may be a more effective means of reducing the risk of spine than hip fractures. PMID- 9356740 TI - The influence of bone morphology on fracture toughness of the human femur and tibia. AB - The influence of porosity, osteon density, osteonal area, osteonal lamellar area, osteon size, and haversian canal size on the tension and shear fracture toughness, that is, the mode I and mode II strain energy release rate (GIc and GIIc), respectively, were investigated for the human femur and the tibia. The results suggest that porosity and osteon density were the best explanatory morphological parameters for GIc and GIIc. Both GIc and GIIc significantly decrease with increasing porosity. They also increase with increasing osteon density, the increase being significant for the femur only. Morphological parameters, altogether, can explain 49%-68% of the variation in fracture toughness. We concluded that, although there must be other factors such as biochemical components and microdamage, osteon morphology has an important influence on fracture resistance of the cortical bone. PMID- 9356741 TI - Case-control study of risk factors for hip fractures in the Japanese elderly by a Mediterranean Osteoporosis Study (MEDOS) questionnaire. AB - A case-control study of hip fracture among the Japanese elderly was carried out in order to assess the risk factors for fractures. On the data obtained from 249 cases and 498 controls matched with ethnicity, sex, age, and residential area, significant risk factors on the lifestyle by multivariate analyses included drinking more than three cups of coffee daily, living in rural areas in the past, sleep disturbance, stroke with hemiplegia, and sleeping in a (Western-type) bed. In contrast, in addition to possession of a large body mass index, moderate alcohol intake and eating fish appeared to be associated with a reduced risk of hip fracture. In conclusion, some traditional Japanese lifestyle characteristics may prevent hip fractures among the Japanese elderly. PMID- 9356742 TI - Assessing quality in primary care. Can we ask the right questions? PMID- 9356743 TI - Improving residents' clinical care and outcomes. Practice Audit Program of the Department of Family Medicine at McMaster University. PMID- 9356744 TI - How public health units fit in. PMID- 9356745 TI - Physician availability and hospital privileges. PMID- 9356746 TI - Breastfeeding and mastitis. PMID- 9356747 TI - Up your nose. PMID- 9356748 TI - Asthma during pregnancy. PMID- 9356749 TI - Ophthaproblem. Embolus. PMID- 9356750 TI - Dermacase. Molluscum contagiosum. PMID- 9356751 TI - Mallet finger. PMID- 9356752 TI - Chronic cough. What's the diagnosis? PMID- 9356753 TI - St. John's wort: an herbal remedy for depression? PMID- 9356754 TI - What factors affect quality of care? Using the Peer Assessment Program in Ontario family practices. AB - OBJECTIVE: To describe the relationship between the quality of care provided by family and general practitioners in Ontario and the demographics of the practitioners. DESIGN: Descriptive study using univariate and multivariate analysis to relate physician demographics to quality of care. SETTING: Ambulatory family and general practices in Ontario. PARTICIPANTS: Each year from 1990 to 1994, all non-specialist physicians in Ontario reaching 70 years of age and a random sample of physicians younger than 70 who had been in practice more than 5 years were selected for assessment. After exclusion criteria were applied, the sample size was 922 physicians. MAIN OUTCOME MEASURES: Grades assigned by the College of Physicians and Surgeons of Ontario's Peer Assessment Committee. RESULTS: Practices were assessed and graded by the Peer Assessment Committee. Grades were related to many variables, but many variables were correlated. Four variables remained significant at the P < .05 level. Younger physicians, female physicians, certificates of the College of Family Physicians of Canada, and urban physicians had, on average, higher grades. CONCLUSION: Grades reflecting quality of care and record keeping were significantly associated with age, sex, certification status, and practice location. These findings should be examined and, for the benefit of patients, methods for improving care should be developed and explored. PMID- 9356755 TI - Self-reported cardiovascular disease and risk factors. Prevalence in Ontario among women 50 and older. AB - OBJECTIVE: To determine the prevalence of self-reported cardiovascular disease and risk factors among Ontario women aged 50 and older. DESIGN: Analysis of the 1990 Ontario Health Survey, a population-based, cross-sectional survey. SETTING: Ontario communities. PARTICIPANTS: Residents of Ontario communities during 1990 who responded to the 1990 Ontario Health Survey (61,239 respondents in 35,479 households), weighted to represent the population of Ontario. MAIN OUTCOME MEASURES: Reported heart disease, hypertension, diabetes, height and weight, physical activity, and smoking habits. RESULTS: Nearly 11% of women aged 50 and older report "heart disease"; 24.9% hypertension, and 5.4% diabetes. Women were less likely than men to smoke daily, to smoke 25 or more cigarettes a day, and to be overweight, but were more likely to have lower levels of physical activity. CONCLUSIONS: The prevalence of self-reported heart disease and medical and lifestyle risk factors for heart disease is relatively high among Ontario women aged 50 and older. Physicians and public health officials must keep women in mind when designing or implementing programs or services for heart disease. PMID- 9356756 TI - Family physicians and sports-injury care. Perceptions of coaches. AB - OBJECTIVE: To describe coaches' education in injury care and management and their club's access to medical care, to describe coaches' perceptions of how family physicians care for sports injuries, and to describe strategies used for overcoming perceived poor advice. DESIGN: A telephone survey using both closed and open-ended questions was conducted. Information was collected as background information to a larger study investigating coaches' decisions about allowing injured athletes to compete. SETTING: All 28 competitive gymnastics clubs in the province of Alberta. The clubs trained athletes for all competitive levels. PARTICIPANTS: All 70 coaches registered with the Alberta Gymnastics Federation as working with female gymnasts were approached; 64 coaches were interviewed. MAIN OUTCOME MEASURES: Injury education, access to medical care, perceptions of sports injury treatment provided by family physicians, strategies employed for overcoming perceived poor advice. RESULTS: Education in injury care and management was varied, as was access to medical care. Direct access to sport specific medical care was available at three of the five elite-level clubs, an arrangement stemming from dissatisfaction with the conventional health care system. At all competitive levels, most coaches were dissatisfied with the recommendations they received from family physicians. Various strategies were employed to acquire more suitable advice. CONCLUSIONS: The results point to a need for improved communication between family physicians and coaches. PMID- 9356757 TI - Screening for cognitive impairment in the elderly. AB - OBJECTIVE: To evaluate the extent and type of screening for cognitive impairment primary care physicians use for their elderly patients, to identify perceived barriers to screening, and to explore whether physicians would be willing to use the clock drawing test as a cognitive screening tool. DESIGN: Mailed questionnaire. SETTING: Primary care practices in the Ottawa-Carleton region. PARTICIPANTS: Family physicians and general practitioners culled from the Yellow Pages and Canadian Medical Directory; 368 of 568 questionnaires were returned for a response rate of 70%. Six respondents had fewer than 30 patients weekly and two responded too late to be included in the analysis; 360 cases were included in the analysis. MAIN OUTCOME MEASURES: Responses to 10 questions on cognitive screening and five on demographics and the nature of respondents' practices. RESULTS: About 80% of respondents reported doing at least one mental status examination during the past year. Only 24% routinely screened patients, although 82% believed screening was needed. Major barriers to cognitive screening were lack of time, risk of offending patients, and possible negative consequences of follow up. Clock drawing was perceived as an acceptable method of screening, if it were proven effective. CONCLUSIONS: Most primary care physicians believe cognitive screening is needed, but few routinely screen their elderly patients. Lack of time is the most important perceived barrier to screening. Primary care physicians are receptive to using the clock drawing test, and, because it is not time-consuming, are less likely to consider lack of time a barrier to testing. The clock test might help bridge the gap between perceived need for screening and actual screening. PMID- 9356758 TI - Long-acting beta 2-agonists. Role in primary care asthma treatment. AB - OBJECTIVE: To examine the efficacy of long-acting beta 2-agonists and their role in primary care asthma management and to review briefly the pharmacology of these agents. QUALITY OF EVIDENCE: Most data presented were derived from randomized, double-blind, placebo-controlled trials. Studies were selected for relevance to asthma management in primary care. MAIN FINDINGS: Long-acting beta 2-agonist use is associated with improvements in both objective and subjective measures of asthma control. At present no evidence suggests that long-acting beta 2-agonists have anti-inflammatory potential. While salmeterol has a longer duration of action than short-acting beta 2-agonists, its onset of action is slower. Salmeterol and formoterol, therefore, should not be used for relief of acute bronchospasm. CONCLUSION: Long-acting beta 2-agonists could be useful for treating asthma in primary care, particularly for controlling symptoms of nocturnal asthma and exercise-induced asthma and for providing convenient maintenance therapy for patients who require regular use of short-acting beta 2 agonists despite concomitant use of optimal doses of inhaled anti-inflammatory medication. PMID- 9356759 TI - Shared mental health care in Canada. Making a joint effort to define our roles. PMID- 9356760 TI - Our strength for tomorrow: valuing our children. Part 3: Child health and the environment. PMID- 9356761 TI - Muscle fatigue and induction of stress protein genes: a dual function of reactive oxygen species? AB - Definitive characterization of the mechanisms of skeletal muscle fatigue is still an area of active investigation. One emerging theory concerns a role for the reactive oxygen species (ROS) produced primarily as a consequence of elevated rates of mitochondrial respiration. It has been theorized that the long-lasting effects of low-frequency fatigue (LFF) can be attributed to disruption of some stage of the excitation contraction coupling (ECC) process. Recent evidence suggests that ROS likely denature one or more proteins directly associated with the sarcoplasmic reticulum (SR) Ca2+ release mechanism. Given the potential of ROS to damage intracellular proteins during subsequent bouts of muscle contractions, the capacity of preexisting antioxidant pathways may be complemented by the synthesis of inducible heat-stress proteins (HSPs). HSPs collectively function to maintain cellular protein conformation during stressful proteotoxic insults. The goal of this article is to illustrate how recent findings suggest a dual role of ROS generated during muscle contractions. PMID- 9356762 TI - The effects of bicycle crank arm length on oxygen consumption. AB - The purpose of this investigation was to determine the effects of various crank arm lengths on oxygen consumption for trained cyclists. Secondary purposes were, if optimal crank arm lengths existed, to determine if these lengths could be predicted based on an individual's leg length. Six trained cyclists completed four experimental protocols riding at a workload of approximately 68% of VO2 max using crank arm lengths of 165, 170, and 175 mm. During each protocol, the cadence, oxygen consumption, and distance traveled were determined, and values were combined to give a VO2.m-1.min-1 value. The values then were placed in either a high, medium, or low efficiency category. Significant differences were found among the three protocols. No significant correlations were found between each subject's most efficient crank arm length and leg length. The results of the study suggest that each subject has a most efficient crank arm length, but it does not appear that optimal crank arm length can be predicted by leg length. PMID- 9356763 TI - A model of oxygen transport capacity changes for independently living older men and women. AB - The purpose of the present investigation was to describe, for a subset of a large random survey of men and women, restricted to the ages of 55 to 85 years, the physiological decay pattern for aerobic fitness and contributing factors of cardiovascular and pulmonary function. The time course of the age-related changes in maximal oxygen uptake (VO2max), ventilatory threshold (TVE), maximal ventilation (VEmax), maximal heart rate (HRmax), and O2 pulse (VO2max/HRmax) were examined by fitting the data to a decaying exponential model by use of a least squares parameter estimation technique. The time constant (tau) was used to describe the rate of decline. The women showed a much slower decline in VO2max (tau = 47.4 years) and TVE (tau = 83.3 years) than the men (tau = 20.8 and 15.4 years, respectively). There was a significant age-related decrease in body weight (0.45 kg.yr-1) in the men, whereas the women showed no change. Pulmonary function did not limit performance based on the very slow decline in VEmax and the normal FEV1.0. The decay in HRmax was better described by a linear model, resulting in an extremely slow tau. Maximal O2 pulse clearly exhibited an exponential decay, with a shorter tau (tau men = 13.5 years; tau women = 28.5 years) than any other variable. PMID- 9356764 TI - The influence of recovery duration on high-intensity exercise performance after oral creatine supplementation. AB - The purpose of this study was to determine the effects of creatine supplementation on the ability to reproduce and maintain a high percentage of peak power output during the second of two bouts of high-intensity cycle sprinting following four different recovery intervals. Eighty healthy, active male subjects were randomly assigned to one of two groups (creatine or placebo) and one of four recovery intervals (30, 60, 90, or 120 s). Two maximal cycle ergometer sprints, separated by the assigned recovery interval were performed before and after a 5-day supplementation protocol in which 20 g/day of creatine (plus 4 g/day glucose) or 24 g/day glucose placebo were ingested by subjects from creatine and placebo groups, respectively. Maximal peak power output (PP) and the absolute time to fatigue (TTF) were compared pre- versus postsupplementation. No significant group interactions were noted in this study. Specifically, creatine supplementation had no effect on subjects' ability to reproduce or maintain a high percentage of PP during the second bout of exercise. PMID- 9356765 TI - The muscle activation-force relationship is unaffected by ischaemic recovery. AB - Since reported changes in muscle activation following fatigue could be affected by alterations in muscle contractile properties, the plantar flexors' activation force relationship was investigated before and following an isometric, intermittent, submaximal fatigue protocol. Voluntary and evoked force and muscle activation was tested pre- and postfatigue with ischaemic and nonischaemic recovery. The muscle activation-force relationship of ischaemic and nonischaemic groups was best described by a second-order polynomial equation with similar y intercepts, slopes, and curvature of the slopes. A significantly increased muscle activation-force slope during recovery may be attributed to decreased muscle activation and not impaired muscle kinetics. The index of muscle activation immediately postfatigue was not significantly different between ischaemic and nonischaemic groups (88.5% vs. 92.7%). No significant difference in the estimate of muscle activation postfatigue with polynomials and interpolated twitch (IT) ratios (superimposed/potentiated doublets) suggested that IT ratios can be used as a general estimate of muscle inactivation following fatigue. PMID- 9356766 TI - Temperature of ingested water and thermoregulation during moderate-intensity exercise. AB - The effect of the temperature of ingested water on the rise in core temperature (Tco) during exercise is not clear. Seven trained subjects were recruited to complete 2 hr of recumbent cycling at 51% VO2peak in a temperate environment (Ta = 26 degrees C, relative humidity = 40%) on four occasions, while ingesting either no fluid (trial NF26), cold water (0.5 degree C; trial CD26), cool water (19 degrees C; trial CL26), or warm water (38 degrees C; trial WA26) during the second hour of exercise. A fifth trial was conducted during which convective and radiative heat loss were reduced by raising Ta to 31 degrees C. During this trial, subjects ingested cold water (0.5 degree C; trial CD31). When compared to WA26, over the second hour of exercise, CD26 attenuated the time-averaged changes in (Tco) and forearm blood flow and decreased whole-body sweat rate and forearm sweat rate (p < .05). Similarly, relative to WA26, the CL26 trial attenuated the time-averaged changes in Tco and reduced whole-body sweat rate (p < .05) during the second hour of exercise, but CL26 had no significant effect on forearm sweat rate or blood flow. Finally, regardless of beverage temperature, water ingestion (vs. NF26) reduced the time-averaged changes in Tco and in heat storage during the second hour of exercise (p < .05). PMID- 9356767 TI - Moderate physical activity can increase dietary protein needs. AB - Six healthy men completed three 1-hr bouts of treadmill walk-jogging at low (L; 42 +/- 3.9% VO2max), moderate (M; 55 +/- 5.6%), and high (H; 67 +/- 4.5%) exercise intensity in order to determine whether moderate physical activity affects dietary protein needs. Both sweat rate and sweat urea N loss were greater (p < .10) with increasing exercise intensity. Seventy-two hour postexercise urine urea N excretion was elevated (p < .05) over nonexercise control (26.6 +/- 2.96 g) with both M (31.0 +/- 3.65) and H (33.6 +/- 4.39), but not L (26.3 +/- 1.86), intensities. Total 72-hr postexercise urea N excretion (urine + sweat) for the M and H exercise was greater than control by 4.6 and 7.2 g, respectively. This suggests that 1 hr of moderate exercise increases protein oxidation by about 29 45 g, representing approximately 16-25% of the current North American recommendations for daily protein intake. These data indicate that the type of exercise typically recommended for health/wellness can increase daily protein needs relative either to sedentary individuals or to those who exercise at lower intensities. PMID- 9356768 TI - Shared mental health care in Canada: a timely document. PMID- 9356769 TI - The nature of generalized anxiety disorder and pathological worry: current evidence and conceptual models. AB - OBJECTIVE: To examine the nature and conceptualization of generalized anxiety disorder (GAD) and chronic worry as well as data bearing on the validity of GAD as a distinct diagnosis. METHOD: Narrative literature review. RESULTS: Although a wealth of data have been obtained on the epidemiology, genetics, and nature of GAD, many important questions remain regarding the validity of current conceptual models of pathological worry and the discriminability of GAD from certain emotional disorders (for instance, mood disorders) and higher-order trait vulnerability dimensions (for example, negative affect). CONCLUSION: Because the constituent features of GAD are salient to current conceptual models of emotional disorders (for example, models that implicate negative affect or worry/anxious apprehension as vulnerability factors), research on the nature of GAD and its associated features should provide important information on the pathogenesis, course, and co-occurrence of the entire range of anxiety and mood disorders. PMID- 9356770 TI - Assessment and treatment of social phobia. AB - Social phobia is an anxiety disorder characterized by heightened fear and avoidance of one or more social or performance situations, including public speaking, meeting new people, eating or writing in front of others, and attending social gatherings. People with social phobia are typically anxious about the possibility that others will evaluate them negatively and/or notice symptoms of their anxiety. Social phobia affects up to 13% of individuals at some time in their lives and is usually associated with at least moderate functional impairment. Research on the nature and treatment of social phobia has increased dramatically over the past decade. As with many of the anxiety disorders, sensitive assessment instruments and effective treatments now exist for people suffering from heightened social anxiety. Typical assessment strategies include clinical interviews, behavioural assessments, monitoring diaries, and self-report questionnaires. Treatments with demonstrated efficacy for social phobia include pharmacotherapy (for example, phenelzine, moclobemide, selective serotonin reuptake inhibitor [SSRI] medications) and cognitive behaviour therapy (CBT) (for example, cognitive restructuring, in vivo exposure, social skills training). Although preliminary comparative studies suggest that both approaches are about equally effective in the short term, each approach has advantages and disadvantages over the other. Trials examining combined psychological and pharmacological treatments are now under way, although no published data on the relative efficacy of combined treatments are currently available. PMID- 9356771 TI - A biopsychosocial understanding of the irritable bowel syndrome: a review. AB - OBJECTIVES: To review and examine the clinical and research literature on irritable bowel syndrome (IBS) with a view to establishing the role that psychiatric factors play in the pathogenesis and treatment of this syndrome. RESULTS: Comorbid psychiatric illness is common with IBS, yet only a small proportion of these patients seek medical attention. Many patients are either satisfied by reassurance or experience symptom relief from medical treatment directed at target symptoms. A small group of patients do not experience much relief, and it is largely this group who come to the psychiatrist's attention. Psychotropic medication is helpful when clinically indicated, and tricyclic antidepressants in small doses (for example, 50 mg) may be helpful for those patients with a pain-predominant pattern of IBS. Psychotherapy (including cognitive, behavioural, relaxation, thermal-biofeedback, insight-oriented therapy, and hypnosis) has been shown to provide relief, although it has often been difficult to differentiate this improvement from a placebo response. CONCLUSIONS: The group of patients with "refractory IBS" used a large amount of health care resources in an attempt to find relief to their distress. Further study is needed to gain a better understanding of which component of psychotherapy is most cost-effective and which patients are most likely to benefit. The large group of those who admit to symptoms compatible with IBS but who do not seek medical attention has to a large extent been excluded from most studies. Exploring this group may provide further insight into this perplexing syndrome. PMID- 9356772 TI - Clinical profile of mania in children and adolescents from the Indian subcontinent. AB - OBJECTIVES: To see whether classic DSM-III-R criteria for mania are applicable to Indian youngsters and to examine the clinical presentation of mania in an Indian child and adolescent psychiatric sample. METHOD: Fifty subjects with a diagnosis of functional psychosis as per the definition in ICD-9 were recruited from the population referred during the study period of approximately one year (n = 840) to the Child and Adolescent Psychiatry (CAP) clinic of the National Institute of Mental Health and Neuro Sciences (NIMHANS), Bangalore, South India. The subjects were systematically evaluated using a standardized clinical interview and demographic questionnaire and were classified according to DSM-III-R. The subjects who satisfied DSM-III-R criteria for mania formed the sample for this study. RESULTS: Twenty-one subjects received a diagnosis of mania according to DSM-III-R. The most common symptoms of mania included pressure of speech, irritability, elation, distractibility, increased self-esteem, expansive mood, flight of ideas, and grandiose delusions. No subject had comorbid attention deficit hyperactivity disorder (ADHD). Additionally, 13 (61%) of the 21 manic subjects had delusions and/or hallucinations. The other common symptoms included psychomotor agitation, reduced sleep, anger, temper tantrums, decreased concentration, disobedience, aggression, and hyperactivity. CONCLUSIONS: Mania was diagnosable in Indian children and adolescents using classic DSM-III-R criteria. The clinical profile appears to be generally similar to that seen in adults. ADHD is not a comorbid condition. The presence of aggressive or disruptive behaviours and hyperactivity in childhood- and adolescent-onset mania, however, could lead to a misdiagnosis of attention-deficit hyperactivity disorder/conduct disorder (ADHD/CD). Similarly, the presence of psychotic features could lead to a misdiagnosis of schizophrenia. PMID- 9356773 TI - Psychiatric admissions of Asian Canadians to an adolescent inpatient unit. AB - OBJECTIVE: To compare the psychiatric diagnoses for Asian Canadians admitted to an adolescent inpatient unit with those of their white Canadian peers. METHOD: A literature review was first completed and then followed by a hospital file review of the Asian Canadians admitted over a 5-year period to the adolescent inpatient psychiatric unit. The data extracted (relating to psychiatric diagnosis, age, length of stay, referral source, family type, and gender) were then compared with a random sample of white Canadians admitted to the same unit during the same 5 year time frame. RESULTS: There were far fewer Asian Canadians admitted than would be expected based on Calgary's demographics. There was equal gender representation among those who were admitted, and they tended to be older and to have a greater preponderance of severe psychiatric symptomatology than their white Canadian peers. CONCLUSIONS: This paper adds to previous research in emphasizing that ethnocultural factors play a significant role in the utilization of psychiatric services by immigrant populations. PMID- 9356774 TI - Expression of depressed mood: a comparative study among Japanese and Canadian aged people. AB - OBJECTIVE: To investigate differences of expression regarding depressed mood between Japanese and Canadian aged people. METHOD: The Zung Self-Rating Depression Scale (SDS) was applied to people aged 65 and over in Ohira, Japan, and Steveston, British Columbia, Canada. RESULTS: The number of subjects who filled out the SDS completely was 2180 for the Japanese sample and 183 for the Canadian sample. The mean SDS indexes of the Japanese and the Canadian samples were 44.03 and 44.34, respectively. The Canadian sample showed a higher average score in 11 items out of 20, whereas the Japanese sample showed a higher score on only 4 items. The factor analysis of those samples showed only small differences. CONCLUSIONS: The Canadian sample showed a higher average score in more items compared with the Japanese sample. This indicates that Canadian aged people express their depressed moods more clearly and spontaneously than Japanese aged people. PMID- 9356775 TI - The incidence of delirium in psychiatric inpatient units. AB - OBJECTIVES: To estimate prospectively the incidence of delirium in psychiatric inpatients and to identify risk factors for delirium in this population. METHOD: The subjects were nondelirious patients newly admitted to the Calgary General Hospital. The Delirium Symptom Interview (DSI), the Confusion Assessment Method (CAM), and the Mini-Mental State Examination (MMSE) were used to identify incident cases of delirium. In order to evaluate the potential impact of selection bias, we conducted a supplementary analysis using record linkage to an electronic administrative data base with coverage of the target population. RESULTS: Of 420 admissions to the hospital, 401 subjects provided informed consent and were not delirious at the time of admission. There were 9 incident cases of delirium. The cumulative incidence rate was, therefore, 2.14 per 100 admissions. The record linkage analysis did not uncover evidence of selection bias. Delirium was associated with a significantly increased length of stay in hospital. CONCLUSIONS: Delirium is an uncommon incident event in the psychiatric inpatient population. The incidence rate reported here may be useful as a benchmark for the identification of excessive rates in other inpatient settings. Since delirium is sometimes related to modifiable therapeutic factors, an excessive rate should prompt a search for its causes. PMID- 9356776 TI - Disorientation in chronic psychiatric patients. AB - OBJECTIVE: To explore the effect of chronic institutionalization on cognitive performance in chronic psychiatric patients with emphasis on age disorientation, a phenomenon that was found in previous research to occur in up to 25% of chronic schizophrenic patients. METHOD: One hundred and ten chronic psychiatric patients, forming 4 main groups--schizophrenic patients, nonschizophrenic patients, institutionalized, and noninstitutionalized--were examined for age disorientation (inability to give one's chronological age correctly on request), and their Minimental State scores (MMSE) were compared across the 4 groups. RESULTS: Twelve out of 43 patients (26%) who were institutionalized according to our definition were age-disoriented and had significantly lower MMSE scores than the other 3 groups. The chronic, noninstitutionalized schizophrenic group and the other chronic psychiatric patients, whether they were institutionalized or not, were negative for this phenomenon. One of the 12 age-disoriented patients was age delusional, and 5 of the 12 had a total MMSE score consistent with dementia (21 or lower). CONCLUSION: Age disorientation is a specific phenomenon that characterizes a subgroup of chronically ill and institutionalized schizophrenic patients. It is unlikely that chronicity per se or prolonged hospitalization alone will lead to cognitive impairment. PMID- 9356777 TI - An investigation of competency to participate in legal proceedings in Canada. AB - OBJECTIVE: To assess fitness to stand trial, competency to plead guilty, and competency to understand Charter cautions to determine if the level of competency varies across these domains. METHODS: The Fitness Interview Test-Revised (FIT-R) and the Test of Charter Comprehension (ToCC) were administered to a group of individuals held on remand for fitness evaluations. Additionally, several questions from the FIT-R that address the ability to make a guilty plea were assessed separately and constituted an individual measure of competency to plead guilty (CoP). RESULTS: As predicted, the results indicated that the fact that an individual is competent at one juncture in the criminal proceedings does not mean that the individual necessarily is competent at all other stages of the proceedings. CONCLUSIONS: These findings suggest a need for a stage-specific approach to forensic competency assessments, requiring specialized instruments designed to assess the legal issues of competency at the various stages of legal proceedings. PMID- 9356778 TI - Re: Clomipramine treatment and behaviour therapy with agoraphobic women. PMID- 9356779 TI - Re: Extrapyramidal side effects with low-dose risperidone. PMID- 9356780 TI - Influence of neutrophil separation on the expression of adhesion molecules. AB - In many assays of polymorphonuclear neutrophil (PMN) function the first step is separation of PMN from whole blood. In the present investigation it was examined if PMN separation leads to an altered expression of neutrophil surface membrane adhesion molecules. Samples have been taken from 20 healthy volunteers (10 male, 10 female; 39.7 +/- 11.8 years of age). PMN activation was measured cytometrically using the following antibodies against PMN surface membrane receptors: L-selectin (CD 62 L), beta-2-integrin Mac-1 (CD 11b) and Intercellular Adhesion Molecule 1 (CD54). PMN activation was determined in whole blood and after separation of PMN using density gradients. After PMN separation all three adhesion molecules appeared increased but the effect was only statistically significant for CD 54 (Wilcoxon test). Data (mean fluorescence intensity in arbitrary units) were: CD 62 L: 62 +/- 37 in whole blood, 82 +/- 28 after separation; p = 0.0674, CD 11b: 94 +/- 55 in whole blood, 111 +/- 47 after separation; p = 0.1454, CD 54; 13 +/- 12 in whole blood, 81 +/- 35 after separation; p < 0.0001. With the present data available it can be assumed that separation of PMN from whole blood can influence the results of flow cytometric assays. PMID- 9356782 TI - Comparison of two methods in erythrocyte microrheology determination using glutaraldehyde-treated cells. AB - The change in human red blood cell microrheology in different glutaraldehyde concentrations (1.5, 3.0 and 5.0 x 10(-6) mol l-1) was studied. The method of millipore filtration was compared with the method of cation-osmotic hemolysis. The obtained results revealed that the prolongation of the erythrocyte filtration time correlated with the curve shifts found in cation-osmotic hemolysis. Contrary to the filtration method, significant differences between two lower concentrations of glutaraldehyde (1.5 and 3.0 x 10(-6) mol l-1) were found. Therefore, we conclude that the cation-osmotic hemolysis is more sensitive than the filtration method in determining the red blood cell deformability. PMID- 9356781 TI - Reduction of myocardial ischemia-reperfusion injury by isovolumic hemodilution. AB - In order to study the effects of isovolumic hemodilution and its combination with Danshen solution on acute ischemic reperfused canine myocardium, 24 adult hybrid dogs were used and divided into four groups. Group I was the control group, groups II-IV were treated with Dextron 40, Danshen solution and a combination of the two, respectively. The results showed that either Dextron 40 or Danshen solution alone had a significant increase of +/- dp/dt-max when compared with group I (p < 0.05). Although no significant difference existed between group II and III, the former showed more rapid action. The combination of the two therapies improved +/- dp/dt-max and LVSP, and significantly reduced the necrotic sizes and the MDA contents in the ischemic myocardia compared with not only group I, but also group II or group III (all p < 0.05). The results suggest that isovolumic hemodilution or Danshen may protect the ischemic reperfused myocardium and the former may come into action more rapidly, and that the combination of the two may show a better synergism than each one of the two by itself. PMID- 9356783 TI - The effect of long-term treatment with hypotensive drugs on blood viscosity and erythrocyte deformability in patients with essential arterial hypertension. AB - This study examined ninety-six patients with essential arterial hypertension (WHO grade I-II) who had been under treatment for a period of at least one year before participating in the study. When the study began the patients received no drug treatment for one month. At the end of this washout period their basal situation was evaluated and drug treatment was begun (26 patients on calcium antagonists, 39 on beta-blockers and 31 on angiotensin converting enzyme inhibitors). The patients were evaluated again after one and two years of uninterrupted treatment with the chosen medication. The results obtained indicate that in a basal situation hypertensive patients have higher blood viscosity and erythrocyte rigidity values than the control group. Regardless of the drug treatment used, the patients experienced during the study a progressive deterioration of their hemorheological situation consisting of an increase in red blood cell rigidity and increased blood viscosity. These results indicate the importance of evaluating the hemorheological parameters of hypertensive patients and suggest that it may be advisable to include in their treatment some kind of medication that prevents progressive rheological deterioration. PMID- 9356784 TI - Erythrocyte, platelet and polymorphonuclear leukocyte membrane dynamic properties in essential hypertension. AB - In subjects with essential hypertension we evaluated, respectively, the red cell membrane protein lateral mobility (obtained marking intact red blood cells with pyrene-3-maleimide (3-PM)), the erythrocyte membrane fluidity (obtained marking intact erythrocytes with 10-(1-pyrene) decanoic acid), the red cell membrane transverse fluidity gradient (obtained marking intact red blood cells with a set of fatty acid fluorescent probes (2-AP, 6-AS, 9-AS, 12-AS)), the platelet membrane fluidity (obtained marking intact and unstimulated platelets with 1,6 diphenyl-1,3,5-hexatriene (DPH) and with 1-(4-(trimethylamino)phenyl)-6-phenyl 1,3,5-hexatriene (TMA-DPH)) and the polymorphonuclear membrane fluidity (obtained marking intact and unstimulated polymorphonuclear cells with TMA-DPH). From the obtained data it is evident that: (1) red cell membrane protein lateral mobility does not distinguish normals from hypertensives; (2) erythrocyte membrane fluidity and red cell membrane transverse fluidity gradient clearly discriminate normals from hypertensives; (3) platelet membrane fluidity differentiates normals from hypertensives only when DPH is used as fluorescent probe; (4) polymorphonuclear membrane fluidity does not distinguish normals from hypertensives. Our results show that in essential hypertension a different behaviour of the membrane dynamic properties in the circulating blood cells is evident. PMID- 9356785 TI - Variations of erythrocyte morphology in different pathologies. AB - In our study we evaluated erythrocytic morphology in different pathologies which can modify flowing red cells. We followed the methodology proposed by Zipursky which allows a three-dimensional evaluation of the red cell and a classification according to the shapes observed through the optical microscope. We studied 150 subjects: 20 normal subjects, 58 patients suffering from vascular diseases, 40 affected by diabetes (type II) (10 without and 30 with vascular diseases), 22 patients with liver disease, 5 patients with monoclonal gammopathies and 5 dehydrated patients. Results show that in normal subjects bowls, which is the shape of the most deformable red cells, are more (55%) than discocytes (44%); the altered forms are only 1%. In vascular patients we noted a statistically significant increase of discocytes (60%). There are no significant differences between subjects affected by diabetes without vascular disease and normal subjects. In diabetics with vascular diseases there are more discocytes (57%) and some altered forms (3%). In patients suffering from chronic hepatitis a great increase (13%) in echinocytes and knizocytes was noticed, which suggests an alteration in the fluidity of the membrane. Our observations testify the importance of this simple methodology in focusing the morphological alterations which can be accounted for both by pathologies of the red cells and by changes in their metabolism. PMID- 9356786 TI - Polymorphonuclear leukocyte membrane fluidity and cytosolic Ca2+ content in different clinical conditions. AB - The aim of the study was to evaluate the polymorphonuclear leukocyte (PMN) membrane fluidity and PMN cytosolic Ca2+ content in several clinical conditions: diabetes mellitus, vascular atherosclerotic disease (VAD), chronic renal failure (CRF), essential hypertension (EH). In 13 subjects with insulin-dependent diabetes mellitus (IDDM), in 24 subjects with non-insulin-dependent diabetes mellitus (NIDDM), in 42 VAD subjects, in 23 VAD subjects with NIDDM, in 15 subjects with CRF and in 12 subjects with EH, we determined the PMN membrane fluidity, obtained marking unstimulated PMN cells with fluorescent probe 1-[4 (trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH), and considering the fluorescence polarization degree, and the PMN cytosolic Ca2+ content, obtained marking unstimulated PMN cells with the fluorescent probe Fura2-AM and considering the ratio between the Fura2-Ca2+ complex and the unchelated Fura 2 fluorescence intensity. From the obtained data it is evident that PMN membrane fluidity does not distinguish normals from IDDM subjects, NIDDM subjects, VAD subjects with and without NIDDM, CRF subjects and hypertensives. PMN cytosolic Ca2+ content, in comparison with normal controls, is significantly increased in VAD subjects (p < 0.01), in VAD subjects with NIDDM (p < 0.001), in CRF subjects (p < 0.001) and in hypertensives (p < 0.05). No correlation was found between PMN membrane fluidity and PMN cytosolic Ca2+ content. The study of these PMN parameters can be useful in the understanding of the role of leukocytes in the vascular damage that characterizes these clinical conditions. PMID- 9356787 TI - Evaluation of the success of hemodilution therapy for fetal growth retardation by Doppler sonography. AB - The aim of our study was to evaluate the success of a hemodilution therapy in patients with intrauterine growth retardation (IUGR) using Doppler sonography. Therefore, 22 patients with IUGR were subjected to hemodilution therapy using infusions of 500 ml hydroxyethylstarch in combination with 500 ml Ringer solution on 14 successive days. The 22 patients were divided into two groups on the basis of the actual birth weight, whereas 13 patients gave birth to an eutrophic infant (AGA group) while 9 infants remained dystrophic (SGA group). The following parameters were determined: hematocrit, prolongation of gestation, pulsatility and resistance index of the umbilical artery, the fetal aorta, the uterine arteries and the middle cerebral artery as well as the fetal outcome. Although the hematocrit decreases in both groups were almost identical, the Doppler sonographic examinations in the AGA group revealed in all cases more favorable improvements in perfusion under therapy in comparison with the SGA group. Considering the fetal aorta we could determine a worsening of perfusion in the SGA group. In the course of our investigations we have found that hemodilution therapy with hydroxyethylstarch represents a promising approach to counteract retarded fetal growth. Doppler sonography progress monitoring appears to be highly suitable for evaluating the response to therapy, especially since it can be assumed that the absence of an improvement in flow indicates only a slight advantage for the child. PMID- 9356788 TI - Brain abscess. AB - The past 20 years have seen major advances in the diagnosis and management of brain abscess, with a corresponding improvement in the survival rates. The advances in radiographic scanning, the availability of new antimicrobials, and the development of novel surgical techniques have all contributed to the decreases in associated morbidity and mortality. The relative rarity of brain abscess and the frequent delays in making the diagnosis render this condition a significant challenge for the clinician. A high index of suspicion is required so that effective therapy can be instituted as soon as possible. Close coordination of care between neurosurgeons and infectious diseases specialists is increasingly important in the complicated management of brain abscess. Adequate abscess drainage and appropriate antimicrobial therapy remain the cornerstones of proper treatment of this condition. PMID- 9356789 TI - Quality standards for immunization. Guidelines from the Infectious Diseases Society of America. AB - This is the third in a series of guidelines commissioned by the Infectious Diseases Society of America through its Practice Guidelines Committee. It is presented as a standard of care rather than a practice guideline because the case for following these recommendations is very strong and it should be followed with rare exceptions. The purpose of this guideline is to provide assistance to clinicians when making decisions on providing immunizations to healthy infants, children, and adults. The document is a summary of guidelines already developed by national organizations. The targeted providers are pediatricians, family practitioners, internists, and other physicians who provide immunizations. The IDSA provided the funding. Panel members represent experts in adult and pediatric infectious diseases. The guidelines are evidence-based. A standard ranking system is used for the strength of the recommendations and the quality of the evidence cited in the literature reviewed. The document has been subjected to external review by peer reviewers as well as by the Practice Guidelines Committee and was approved by the IDSA Council. An executive summary and tables highlight the major recommendations. Indicators for measuring compliance with the standards are included. PMID- 9356790 TI - Practice guidelines for community-based parenteral anti-infective therapy. ISDA Practice Guidelines Committee. AB - This is the fourth in a series of practice guidelines commissioned by the Infectious Diseases Society of America through its Practice Guidelines Committee. The purpose of this guideline is to provide assistance to clinicians when making decisions on when and how to best administer parenteral antimicrobial therapy. The targeted providers are internists, pediatricians, family practitioners, and other providers of outpatient antiinfective therapy. Criteria for selecting the appropriate patients and settings to deliver therapy in the community are described. Panel members represented experts in adult and pediatric infectious diseases. The guidelines are evidence-based. A standard ranking system is used for the strength of the recommendations and the quality of the evidence cited in the literature reviewed. The document has been subjected to external review by peer reviewers as well as by the Practice Guidelines Committee and was approved by the IDSA Council. An executive summary and tables highlight the major recommendations. PMID- 9356791 TI - Human immunodeficiency virus/AIDS Medicaid managed care network. PMID- 9356792 TI - Molecular fingerprinting of Mycobacterium tuberculosis: how can it help the clinician? AB - In just a few years, molecular fingerprinting of Mycobacterium tuberculosis has provided clinicians with significant insight into the epidemiology of tuberculosis. This methodology has allowed for a new understanding of the extent of new transmission of tuberculosis among residents of various communities and within institutions. It has also allowed for differentiation between episodes of reinfection and relapse, a task hitherto almost impossible to accomplish. In addition, molecular fingerprinting has allowed assessment of situations where laboratory cross-contamination is suspected. Thus, this technology has in many ways made clinicians reexamine many of their long-held beliefs regarding tuberculosis. In this report, Drs. Behr and Small provide a lucid description of molecular fingerprinting of M. tuberculosis, its current uses, and its future potential value. PMID- 9356793 TI - Differential prognosis of gram-negative versus gram-positive infected and sterile pancreatic necrosis: results of a randomized trial in patients with severe acute pancreatitis treated with adjuvant selective decontamination. AB - Results of a previous randomized multicenter trial involving 102 patients with severe acute pancreatitis treated with or without adjuvant selective decontamination (SD) were analyzed additionally with regard to the bacteriologic status of (peri)pancreatic necrosis. The incidence of gram-negative pancreatic infection was significantly reduced in patients treated with SD (P = .004). Once such an infection develops, mortality increases 15-fold (P < .001) in comparison with that for patients with sterile necrosis. Among patients in whom only gram positive infection of pancreatic necrosis was found, there was no significant increase in mortality. These results were similar in both treatment groups. In addition, the hospital stay was significantly longer in cases of gram-negative infected necrosis. The incidence of gram-positive infected necrosis in patients treated with SD did not increase. Gram-negative pancreatic infection can be prevented with adjuvant SD, thereby reducing mortality among patients with severe acute pancreatitis. PMID- 9356794 TI - Selective decontamination of the digestive tract in acute severe pancreatitis--an indication whose time has come. PMID- 9356795 TI - Streptococcus bovis infection of the central nervous system: report of two cases and review. AB - Streptococcus bovis is an uncommon cause of meningitis and subdural empyema. We report one case each of meningitis and subdural empyema in which S. bovis biotype II was isolated from both the spinal fluid and blood. In one case, the organisms were seen on a gram-stained preparation of cerebrospinal fluid. The first patient presented with gastrointestinal symptoms of unknown etiology, was immunosuppressed, and recovered. The second patient presented with syncope, developed a subdural empyema, and died; at autopsy, a colonic adenoma was found. A review of the English-language literature revealed only 14 previously reported cases of meningitis due to S. bovis and no cases of subdural empyema due to S. bovis. These cases indicate the importance of complete laboratory identification of specific organisms and confirm the need for a thorough neurological examination and search for underlying gastrointestinal disease in cases of S. bovis infection. PMID- 9356796 TI - Effectiveness of clemastine fumarate for treatment of rhinorrhea and sneezing associated with the common cold. AB - Limited data support the use of first-generation antihistamines for treatment of the common cold. The purpose of this study was to test the effectiveness of clemastine fumarate, a first-generation antihistamine, for treatment of sneezing and rhinorrhea associated with naturally occurring common colds. Four hundred three subjects (202 clemastine fumarate recipients and 201 placebo recipients) who reported new onset (< 24 hours) of cold symptoms that included rhinorrhea or sneezing were studied. At baseline (day 1), the mean symptom-severity scores +/- SEM for the clemastine fumarate and placebo groups were not significantly different. The mean rhinorrhea-severity score +/- SEM was not different on day 2; however, on day 3, the mean rhinorrhea-severity score +/- SEM was 1.02 +/- 0.07 for the clemastine fumarate group and 1.39 +/- 0.07 for the placebo group (P < .001). This treatment effect persisted on day 4. A significant effect on sneezing was noted on days 2-4. Sedation occurred in 14% of the clemastine fumarate treated subjects and 1.5% of the placebo-treated subjects (P < .0001). PMID- 9356797 TI - Yersinia enterocolitica: a cause of chronic polyarthritis. AB - To investigate the role of Yersinia persistence in chronic undifferentiated arthritis, two patients who had chronic undifferentiated polyarthritis and circulating IgA and IgG antibodies to Yersinia outer proteins were studied. Immunofluorescence using antibodies directed against Yersinia adhesin A was performed on colonic and synovial tissue. Synovial tissue T cells were cloned aspecifically and screened for their proliferative responses to Yersinia enterocolitica. Furthermore, a Yersinia-specific polymerase chain reaction (PCR) was performed on synovial tissue. Both patients were found to have Yersinia antigens in colonic and synovial tissue. Y. enterocolitica-positive T-cell clones were grown from the synovial tissue: 4 CD4+ clones of 37 clones from patient 1 and 6 CD4+ clones of 53 clones from patient 2. Yersinia-specific PCR products were not detected in the synovial tissue specimens. The results support the hypothesis that an immune-mediated response to Yersinia antigens may play an important role in the pathogenesis of chronic undifferentiated arthritis. PMID- 9356798 TI - Four additional cases of Burkholderia gladioli infection with microbiological correlates and review. AB - Burkholderia gladioli has only recently been reported to be a human pathogen. Four cases of B. gladioli infection (including bacteremia, pneumonia, and cervical adenitis) in two adults and two young children are reported. Three of these four patients were severely immunocompromised. Commercial systems were frequently unable to identify this bacterium correctly. Antimicrobial susceptibility patterns indicated that B. gladioli strains were susceptible to the quinolones, aminoglycosides, and imipenem. In vitro laboratory investigations demonstrated that B. gladioli strains were susceptible to complement-mediated lysis of pooled human serum, thus implying that healthy individuals should be immune to infection. These four cases together with three previously reported cases suggest that B. gladioli primarily causes disease in severely immunocompromised individuals. The lack of mortality associated with infection, coupled with susceptibility to serum and lack of recognizable virulence associated factors, suggests that this species has a low pathogenic potential. PMID- 9356799 TI - Point prevalence of oropharyngeal carriage of fluconazole-resistant Candida in human immunodeficiency virus-infected patients. AB - To estimate the prevalence of both clinically evident and asymptomatic carriage of fluconazole-resistant Candida, we prospectively surveyed 128 adults infected with human immunodeficiency virus (HIV). The patients had an average CD4 cell count of 206/mm3. Ninety-seven isolates of Candida were obtained from the oropharynx of 82 patients (64%). Of these 82 patients, 76% carried C. albicans alone; 18%, both albicans and non-albicans isolates; and 6%, non-albicans species alone. Oropharyngeal candidiasis was evident in only 38 (46%) of the 82 patients for whom a culture was positive and was never seen unless C. albicans was present. When MICs were measured by using the National Committee for Clinical Laboratory Standards M27-T methodology and grouped by using recently proposed breakpoints, we found that eight of the 38 patients with oropharyngeal candidiasis and six of the 44 patients who were asymptomatically colonized carried C. albicans isolates resistant to fluconazole (MIC, > or = 64 micrograms/mL); estimated rates of carriage were 21% (95% confidence interval, 10%-37%) and 14% (95% confidence interval, 5%-27%), respectively. Carriage of resistant isolates of C. albicans by HIV-infected adults is more common than previously suspected, and clinicians should be alert to the possible need for either higher doses of fluconazole or alternative treatment modalities. PMID- 9356800 TI - Neurological complications of chlamydial infections: case report and review. AB - We describe a patient with Chlamydia pneumoniae infection who presented with cerebellar dysfunction, followed by respiratory failure requiring mechanical ventilation. C. pneumoniae is an important respiratory pathogen, and other clinical manifestations, including neurological syndromes, are being increasingly recognized. Meningoencephalitis and other neurological complications have also been described in patients with infections due to Chlamydia psittaci and Chlamydia trachomatis. Chlamydial infections should be included in the differential diagnosis of neurological syndromes, including cerebellar dysfunction. PMID- 9356801 TI - Risk factors for meningitis after transsphenoidal surgery. AB - To evaluate possible risk factors for meningitis, we retrospectively reviewed 228 transsphenoidal operations (in which a standard regimen of amoxicillin prophylaxis was used) for sellar pathology. The incidence of meningitis was 3.1% (seven of 228 cases). Cultures of preoperative specimens from the anterior nasal vestibule in three of seven patients yielded Staphylococcus aureus, but none of these patients developed S. aureus meningitis. Two of three patients with significant preoperative paranasal sinus abnormalities developed meningitis compared with only five of 225 patients without significant paranasal sinus abnormalities (P < .005). Three of 22 patients with intraoperative cerebrospinal fluid (CSF) leakage developed meningitis compared with four of 206 patients without intraoperative CSF leakage (P < .05). Six of seven patients with postoperative CSF rhinorrhea and only one of 221 patients without postoperative CSF rhinorrhea developed meningitis (P < .00001). In conclusion, postoperative CSF leakage is an important risk factor for meningitis after transsphenoidal surgery. Cultures of preoperative specimens from the anterior nasal vestibule did not have any predictive value in our study. PMID- 9356802 TI - The clinical microbiology laboratory and infection control: emerging pathogens, antimicrobial resistance, and new technology. AB - The clinical microbiology laboratory is an essential component of an effective infection control program. Laboratory personnel have a broad range of technologies, from traditional methods of detecting and identifying organisms to modern molecular typing methods, that they can use to support and enhance the efforts of the infection control staff. If the infection control team applies these technologies appropriately, it can prevent problems and solve nosocomial mysteries efficiently. In this era of cost-containment, staff members in the laboratory and in the infection control program must work hard to communicate their unique and shared goals, needs, and problems. If the laboratory and infection control personnel cooperate and collaborate rather than compete, both programs will be successful and the patients and the hospital will benefit because the risk of nosocomial infections and the frequency of resistant organisms will be reduced. PMID- 9356803 TI - Adjunctive corticosteroid therapy for tuberculosis: a critical reappraisal of the literature. AB - An extensive, although largely forgotten, literature addresses the utility of adjunctive corticosteroid therapy in the management of tuberculosis. Corticosteroid therapy probably improves neurological outcomes of, and decreases mortality due to, tuberculous meningitis of moderate severity. Although therapy for tuberculous pericarditis is simplified (with less need for operative intervention) by adjunctive corticosteroid administration and there are fewer deaths, the incidence of subsequent constriction is not changed. The signs and symptoms of typical reactivation tuberculous pneumonia, tuberculous pleurisy, and probably primary tuberculous disease (with lymphadenopathy) seem to decrease rapidly with corticosteroid therapy, although no differences in final outcomes have been observed. Corticosteroid regimens used in most studies varied greatly in duration and dosage and generally caused significant side effects. Corticosteroids do not appear to diminish the efficacy of adequate antimycobacterial therapy. Adjunctive corticosteroid therapy appears to offer significant short-term but (other than for tuberculous meningitis and effusive pericarditis) minimal long-term benefit for patients with tuberculosis. PMID- 9356804 TI - Iron, infections, and anemia of inflammation. AB - Iron is essential to all microorganisms. To obtain iron from the very low concentrations present in their environment, microorganisms have developed sophisticated mechanisms such as the siderophore system. As a primitive defense mechanism, humans have developed mechanisms to withhold iron from microorganisms. Iron-binding proteins such as transferrin, ferritin, and lactoferrin have a central role in human ferrokinetics. These iron-binding proteins also participate in the process of decreasing iron availability for the microorganisms. They do so by decreasing iron reutilization. Anemia of inflammation (previously called anemia of chronic disease) is seen in the setting of infectious, inflammatory, and neoplastic diseases. It results, in part, from changes in the intracellular metabolism of iron. Alterations of iron physiology seen in many clinical circumstances make excess iron available to microorganisms, thus enhancing their pathogenicity. Understanding the molecular basis of iron withholding by the human host, both in the absence of and during infection, and that of iron acquisition by microorganisms may provide us with new and innovative antimicrobial agents and vaccines. PMID- 9356805 TI - Pneumonia associated with near-drowning. AB - Drowning and near-drowning can abruptly devastate the lives of both the affected victims and their families. In addition to the complications directly caused by the submersion, several indirect causes of morbidity exist. Infection is one of the complications associated with near-drowning, and pneumonia is the most severe of these infectious complications. The risk factors, microbiological causes, diagnostic approach, and appropriate therapy for pneumonia associated with near drowning are not well described in the literature. Herein, we review the epidemiology and pathophysiology associated with near-drowning, discuss the potential mechanisms of infection, and describe the likely risk factors for pneumonia related to near-drowning. We also detail the microbiological causes of this entity and provide important clinical and epidemiological information associated with specific pathogens. Finally, we summarize an appropriate diagnostic and therapeutic approach to pneumonia associated with near-drowning. PMID- 9356806 TI - Development of fluconazole resistance in Candida albicans causing disseminated infection in a patient undergoing marrow transplantation. AB - Oral candidiasis due to azole-resistant Candida albicans is an increasing problem in patients with AIDS who received prolonged periods of fluconazole prophylaxis. Infection with C. albicans is also frequent in patients undergoing transplantation. However, azole resistance has not been appreciated as a major problem for these patients, presumably because they receive a relatively short duration of fluconazole prophylaxis. We describe a case of disseminated candidiasis due to fluconazole-resistant C. albicans in a patient following marrow transplantation. Restriction fragment length polymorphism analysis with use of the C. albicans strain-specific Ca3 probe was performed on sequential isolates. Identical banding patterns were obtained, thereby confirming that a fluconazole-susceptible endogenous C. albicans acquired azole resistance during a brief exposure to the drug and subsequently caused disseminated infection. This observation raises questions regarding the incidence, significance, and mechanism of azole resistance in fungi causing infection in this population. PMID- 9356807 TI - Progressive vaccinia treated with ribavirin and vaccinia immune globulin. AB - A 67-year-old man with metastatic melanoma and chronic lymphocytic leukemia was inadvertently given a vaccinia melanoma oncolysate vaccination. He developed progressive vaccinia at the site of inoculation. The lesion started to heal only when he was treated with ribavirin. Vaccinia immune globulin was administered and appeared to help control the initial lesion and limit the development of satellite lesions. PMID- 9356808 TI - Invasive Haemophilus influenzae type b (Hib) infection in an adult patient with a selective deficiency of antibody to the Hib capsular polysaccharide. AB - A 31-year-old woman without any underlying disease contracted severe invasive Haemophilus influenzae type b (Hib) infection but developed no antibodies to the Hib capsular polysaccharide. Serum immunoglobulin levels were normal, but she had an isolated deficiency of antibody to Hib. Subsequently, immunization with a tetanus toxoid-conjugated Hib vaccine induced only a minimal response. However, she had a protective level of antibody (> 1 microgram/mL) after the fifth vaccination. PMID- 9356809 TI - Successful treatment of Edwardsiella tarda prosthetic valve endocarditis in a patient with AIDS. PMID- 9356810 TI - An unusual case of renal abscess caused by Streptococcus pneumoniae. PMID- 9356811 TI - Pasteurella multocida prosthetic valve endocarditis: case report and review. PMID- 9356812 TI - Phaeohyphomycosis following cardiac surgery: case report and review of serious infection due to Bipolaris and Exserohilum species. PMID- 9356813 TI - Delayed parasite clearance in a splenectomized patient with falciparum malaria who was treated with artemisinin derivatives. PMID- 9356814 TI - Primary septic arthritis and osteomyelitis due to Mycobacterium avium complex in a patient with AIDS. PMID- 9356816 TI - Detection of serum antibodies to Chlamydia pneumoniae in patients with endogenous uveitis and acute conjunctivitis. PMID- 9356815 TI - Microsporidium species in pulmonary cavitary lesions of AIDS patients infected with Rhodococcus equi. PMID- 9356818 TI - Cervical cytomegalovirus infection in a woman with AIDS. PMID- 9356817 TI - A semiquantitative analysis of the fecal flora of patients with vancomycin resistant enterococci: colonized patients pose an infection control risk. PMID- 9356820 TI - Nocardia farcinica pneumonia in a previously healthy woman: species characterization with use of a digoxigenin-labeled cDNA probe. PMID- 9356819 TI - Unusual prolonged hypocalcemia due to foscarnet in a patient with AIDS. PMID- 9356821 TI - Rapidly fatal infection due to Photobacterium (Vibrio) damsela. PMID- 9356822 TI - Successful treatment of multidrug-resistant Pseudomonas aeruginosa meningitis with high-dose ciprofloxacin. PMID- 9356823 TI - Hypertrophy of the breasts in a patient treated with indinavir. PMID- 9356824 TI - Use of the polymerase chain reaction for the detection of Legionella pneumophila DNA in serum samples. PMID- 9356826 TI - Lancefield serogrouping alone is insufficient for species assignment of streptococci. PMID- 9356825 TI - Problems with antibiotic resistance in Spain and their relation to antibiotic use in humans elsewhere. PMID- 9356827 TI - Enzyme-linked immunosorbent assay, not agglutination, is the test of choice for the diagnosis of neurobrucellosis. PMID- 9356828 TI - Detection of laboratory cross-contamination of Mycobacterium Mycobacteriu cultures. PMID- 9356829 TI - Primary amebic meningoencephalitis in a patient with AIDS: unusual protozoological findings. PMID- 9356831 TI - Preface to the 1997 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with human immunodeficiency virus. USPHS/IDSA Prevention of Opportunistic Infections Working Group. US Public Health Service/Infectious Diseases Society of America. PMID- 9356830 TI - Emerging antibiotic resistance in Indian communities. PMID- 9356832 TI - 1997 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with human immunodeficiency virus: disease-specific recommendations. USPHS/IDSA Prevention of Opportunistic Infections Working Group. US Public Health Services/Infectious Diseases Society of America. PMID- 9356833 TI - Radiation therapy for primary gallbladder cancer. AB - This paper presents a survey into the role of radiation therapy in the treatment of advanced gallbladder cancer and provides information on the radiobiological aspects for combining surgery and radiation therapy. This treatment modality has resulted in improved median survival time even in patients with stage IV disease when it was applied to microscopic post-resection residue. PMID- 9356834 TI - Staging of carcinoma of the gallbladder--an ultrasonographic evaluation. AB - BACKGROUND/AIMS: Preoperative staging of carcinoma of the gallbladder is important in choosing the most appropriate treatment modality. Ultrasonography is commonly used for the diagnosis of carcinoma of the gallbladder, but its role in staging of the disease has not been adequately evaluated. The aim of this study was to prospectively evaluate the role of ultrasonography performed by a single operator in the staging of carcinoma of the gallbladder. METHODOLOGY: Twenty-six patients with carcinoma of the gallbladder were evaluated in this study. Ultrasonographic staging was done by a single senior radiologist and compared with surgical staging using the AJCC TNM staging system. A retrospective evaluation was done of 122 operated patients with carcinoma of the gallbladder, and the findings were compared to the results of the prospective study. RESULTS: In the prospective study, the overall accuracy of ultrasonogaphy in correctly staging the disease was 38% (10/26). Only 1 patient was overstaged, while the majority of patients were understaged (15) due to missed distant metastasis (10) and missed advanced local infiltration (5). The sensitivity in detecting liver infiltration was 50%; lymph node metastasis, 50%; and liver metastasis, 8%. In the retrospective study, the similar detection rates were 27%, 25% and 10%, respectively. CONCLUSION: Real time ultrasonography has been found to have inherent limitations in staging carcinoma of the gallbladder. Its accuracy, though improved in a prospective study, is inadequate in staging the disease accurately. More accurate imaging modalities are therefore required to accurately stage the disease preoperatively, so as to avoid unnecessary laparotomies. PMID- 9356835 TI - Factors predictive of early complications of endoscopic treatment of bile duct calculi. AB - BACKGROUND/AIMS: Factors associated with an increased early complication rate of the endoscopic sphincterotomy procedure have been identified. Precut or needle knife papillotomy has been shown to improve the success rate of endoscopic retrograde cholangiography and endoscopic sphincterotomy, but has often been reported to be hazardous. In order to identify patients with bile duct stones at risk for a complicated course in connection with endoscopic clearance of the calculi, factors predictive of early complications were sought. METHODOLOGY: 417 consecutive patients with bile duct calculi at endoscopic retrograde cholangiography were considered for endoscopic treatment in our department from 1981 to 1992. Endoscopic sphincterotomy was performed in 246 patients with intact gallbladders and in 147 with prior cholecystectomy, 55 of whom had retained calculi. RESULTS: There was a 9.4% overall and 7.1% major early complication rate of the EST procedure and a 30-day mortality of 0.5% (2 patients, non-procedure related). In 22% (6/27) of the patients with major complications, surgery was required or preferred to additional endoscopic measures. Complete stone removal failed in 35/393 patients (8.9%). The immediate and early complication rate of standard sphincterotomy was not found to be increased in patients with prior or present biliopancreatic complications, failed bile duct clearance at first attempt, or juxtapapillary diverticula. It was the same after standard sphincterotomy as after precut papillotomy followed by immediate or delayed sphincterotomy. No increased morbidity was found after failed therapy as compared to failed diagnostic precut papillotomy. There was neither a greater need for, nor an increased complication rate following, precut papillotomy in patients with, as compared to those without, juxtapapillary diverticula. Endoscopic experience did not influence the complication rate. There were no significant differences regarding outcome or risk factors associated morbidity between patients with and without intact gallbladder. CONCLUSIONS: These findings confirm that endoscopic treatment is safe and that precut papillotomy can be performed without increased morbidity. Furthermore, none of the commonly identified factors associated with increased morbidity were found to be risk factors in this study. PMID- 9356836 TI - Carcinoma of the extrahepatic bile duct: mode of spread and its prognostic implications. AB - BACKGROUND/AIMS: The postoperative course of patients with bile duct carcinoma after surgical resection remains dismal. The purpose of this study was to examine the mode of spread from the original site of the carcinoma and its prognostic significance. METHODOLOGY: A total of 46 Japanese patients with extrahepatic bile duct carcinoma who underwent surgical resection from January 1976 to August 1995 were retrospectively reviewed. RESULTS: Out of 24 patients with upper bile duct carcinoma, 16 (67%) were papillary or well differentiated tubular adenocarcinoma of the polypoid or nodular type on gross configuration, whereas 7 of 11 patients (64%) with lower bile duct carcinoma had moderately differentiated tubular adenocarcinoma or poorly differentiated adenocarcinoma of the annular constrictive or diffusely infiltrating type (p < 0.01). A noteworthy feature was perineural invasion (18/24, 75%) in the former group. Lymphatic permeation and venous invasion were seen in 50% and 38% of the former group and these were present in 73%, and in 73% of the latter (p < 0.01). Lymph node metastasis was most frequent in patients with lower bile duct carcinoma (5/11, 45%). Periductal spread along the bile duct toward the liver was more frequent and extensive in the infiltrating type than in the polypoid, nodular, or annular constrictive type. Carcinoma of the polypoid type often extended along the mucosa and rarely through the periductal layer. The mean distance between the edge of carcinoma invasion estimated by cholangiography and that proved by histology on the resected specimens was 6.1 +/- 6.1 mm in the hepatic and 6.2 +/- 9.1 mm in the duodenal direction. In all 11 patients with lower bile duct carcinoma, surgical margins were free of cancer cells, while they were affected by malignant cells in 17 of 24 patients with upper bile duct carcinoma. Univariate log-rank analysis showed that venous invasion, perineural infiltration, and the presence or absence of cancer cells at the cut edge of the bile duct in the hepatic direction and at the resection margins in the transverse direction were significant prognostic factors. Multivariate Cox regression analysis revealed that cancer cells at the edge of the bile duct in the hepatic direction constitute a significant and independent prognostic variant. CONCLUSIONS: Extrahepatic bile duct carcinoma should be excised to a distance of 1.5 cm from the edge of the carcinoma as estimated on cholangiography to achieve cancer-free margins, especially at the resected margins in the hepatic direction. PMID- 9356837 TI - Multiple hepatic cysts along the intrahepatic bile duct--case report. AB - A case of multiple hepatic cysts of the periductal gland located along the left intrahepatic bile duct is described. Ultrasonography and computed tomography disclosed many cystic lesions along the left portal vein in the left lateral segment of the liver. Percutaneous transhepatic cholangiography showed many compressed lesions. The resected specimen revealed multiple cysts of 2-7mm in diameter along the intrahepatic bile duct. Microscopically, cysts within the large Glisson's capsule were intermixed with lobuli of the periductal glands, thus suggesting periductal gland origin. Histopathological features of these cysts were similar to those of "multiple hilar cysts" reported by Nakanuma, but lack of portal hypertension and underlying chronic liver disease is the significant characteristics in this case which is different from "multiple hilar cysts". PMID- 9356838 TI - Cholangitis after endoscopic biliary drainage for hilar lesions. AB - BACKGROUND/AIMS: Cholangitis is a morbid complication following endoscopic biliary drainage. This study was undertaken to determine the subgroup of patients more likely to develop cholangitis after endoscopic drainage and to determine how to manage this complication. METHODOLOGY: Although we have used transhepatic biliary drainage as the procedure of choice for relieving obstructive jaundice, we recently encountered ten patients who underwent endoscopic drainage before referral. RESULTS: Three patients with a hilar obstruction developed cholangitis and cholangiolytic liver abscesses after endoscopic drainage, while seven with a distal obstruction did not develop cholangitis following drainage. Thus, the incidence of cholangitis in patients with a hilar obstruction (100%) was significantly higher than that (0%) in patients with a distal obstruction (p = 0.008). Cholangitis in the three patients was managed with percutaneous drainage without mortality, although this procedure was technically difficult because of the presence of collapsed intrahepatic bile ducts following the endoscopic drainage. CONCLUSIONS: Hilar lesions present a higher risk of cholangitis after endoscopic biliary drainage than distal lesions. Hence, percutaneous, rather than endoscopic, drainage is indicated for such lesions. Percutaneous drainage is the procedure of choice for the management of cholangitis after endoscopic drainage. PMID- 9356839 TI - Studies on reoperations of extrahepatic biliary tree. AB - Cholelithiasis is one of the most common health disorders, but biliary surgery is burdened with numerous specific complications. These reoperations may be divided into early and late reoperations in terms of their precise therapeutic and prognostic differentiation. Recently, residual stones have been successfully treated by endoscopic sphincterotomy and percutaneous or trans-T-tube tract choledochoscopy, and reoperation is not frequently required. The purpose of this study was to identify the most common complications of biliary surgery which require reoperation so as to establish the predisposing factors and suggest alternative strategies. Thus a retrospective review of biliary surgery at the Medical University of Varna, Bulgaria was performed to identify those cases which required reoperation for complications of biliary surgery and the specific complications which required additional surgery. Of 2874 biliary operations for benign conditions, 87 reoperations were performed: 34 due to early complications and 53 due to late complications. Forty-nine of the 87 patients had undergone the initial surgery in our hospital, while 38 had been in different hospitals. Residual stones were the cause for reoperation in 37 patients. Other causes were: stenosing papillitis: 22 (with or without stones and cholangitis); iatrogenic injuries (5); strictures (5); pancreatitis (7); fistulae, abscesses, and bleeding. The most commonly performed procedure during reoperation was biliodigestive anastomosis (36), followed by T-tube drainage (28), papillosphincteroplasty (4), and double drainage (PSP + BDA), which was performed in 4 patients. In order to avoid reoperation after biliary surgery, exact preoperative diagnosis (by ERCP or US), intraoperative cholangiography and choledochoscopy and accurate operative technique are of great importance. PMID- 9356840 TI - Ceftriaxone in the treatment of spontaneous bacterial peritonitis: ascitic fluid polymorphonuclear count response and short-term prognosis. AB - BACKGROUND/AIMS: In this study, ascitic fluid polymorphonuclear (PMN) response, short-term prognosis, and factors related to hospital mortality were investigated in 62 cases of spontaneous bacterial peritonitis occurring in cirrhotic patients treated with Ceftriaxone (1g every 12 hours). METHODOLOGY: The diagnostic criteria for (SBP) were ascitic fluid PMN count < 250 cells/mm3 and no evidence of secondary peritonitis. Analysis of ascitic fluid samples were obtained on admission, and on the 4th and 10th days of antibiotic therapy. RESULTS: The etiology of cirrhosis was alcohol in 63% of the cases, and 79.5% of patients belonged to Child-Pugh Class C. Ascitic fluid analysis showed positive cultures in 47% of the cases, and a marked decrease in PMN count during treatment (admission: 7762 +/- 2837; 4th day: 388 +/- 91; 10th day: 173 +/- 59 cells/mm3). Ascitic PMN was < 250 cells/mm3 within 4 days of treatment in 33% of the cases. The hospital mortality rate was 24%, and was related to gastrointestinal hemorrhage, hepatic encephalopathy, renal failure and 4th day ascitic fluid PMN count. CONCLUSION: Ceftriaxone is a safe and effective option for the treatment of SBP. PMID- 9356841 TI - An experimental study of hepatic resection using an in situ hypothermic perfusion technique. AB - BACKGROUND/AIMS: Recently, along with the progression of hepatic surgery, the in situ hypothermic perfusion technique has been used for major hepatic resection. The aim of this study was to elucidate the utility of the hypothermic perfusion technique in hepatic resection. METHODOLOGY: Two experimental models: a Total Vascular Occlusion group (TVO) and a Total Vascular Occlusion under Hypothermic Perfusion group (HP) were utilized using adult mongrel dogs. An approximately 40 45% hepatic resection was performed under total vascular occlusion with a systemic and portal shunt. In the HP group, the liver was perfused through the portal vein with lactated ringer's solution at 4 degrees C. In the TVO group, hepatic resection was done without perfusion. RESULTS: Serum AST and ALT levels after reperfusion were significantly increased in the TVO group. From the change of serum hyaluronic acid and xanthine oxidase activity, sinusoidal endothelial cell function was maintained in HP more than in TVO. The mean arterial pressure, portal venous pressure and portal venous blood flow were maintained in HP after reperfusion. Examination of vascular permeability using monastral blue showed that vascular permeability of the small intestine, lung and liver was clearly increased in TVO. Chemiluminescense intensity of the hepatic venous blood after reperfusion gradually increased only in TVO. In addition, the chemiluminescense intensity of the hepatic venous blood congested in the liver increased markedly as vascular occlusion continued in TVO. The hepatic venous blood serum congested in the liver induced morphological changes in the human umbilical vein endothelial cells and increased their permeability. The SDS-PAGE of the hepatic venous blood serum congested in the liver revealed several proteins between 37 and 42 KD. CONCLUSION: The hypothermic perfusion technique in hepatic resection may be very useful in preserving the hepatocytes and sinusoidal endothelial cells and in maintaining stability of the systemic or hepatic circulation after reperfusion because of the cooling of the liver and the washing out of congested blood in the liver. PMID- 9356842 TI - Class I HLA antigens hepatic display and beta-2-microglobulin serum values in chronic hepatitis C: effect of treatment with recombinant alpha interferon. AB - BACKGROUND/AIMS: Enhanced hepatocellular display of class I HLA antigens together with rising serum beta-2-microglobulin (a subunit of class I HLA molecule) and transaminases is reported in patients with chronic hepatitis B during treatment with interferon as an index of immune lysis of virus infected cells. METHODOLOGY: We studied class I HLA antigens and beta-2-microglobulin display in the livers of 23 patients with chronic hepatitis C before and after a 12 month treatment with recombinant alpha interferon. Beta-2-microglobulin serum values were monitored. In all the patients before treatment, class I HLA antigens and beta-2 microglobulin were diffusely displayed in the bile duct epithelium, in the sinusoidal lining cells, in approximately 50% of the inflammatory cells and in the hepatocyte membrane with marked staining in the areas of periportal and lobular necrosis. RESULTS: At the end of the treatment, class I HLA antigens and beta-2-microglobulin were no longer or only faintly detectable in the hepatocytes of 12 patients who showed clinical and histological improvement. The immunohistochemical pattern was unchanged in the 11 patients who did not respond to the therapy. Baseline serum beta-2-microglobulin values were high in all the patients and decreased significantly only in the group of responders. No peaks of transaminases were registered. CONCLUSIONS: The disappearance or reduction of HLA hepatocellular display without acute increase of serum beta-2-microglobulin values and transaminases during successful treatment with interferon in chronic hepatitis C suggests a clearance of the virus due to direct antiviral rather than immunologically mediated mechanism. PMID- 9356843 TI - Interstitial laser thermotherapy in pig liver: effect of inflow occlusion on extent of necrosis and ultrasound image. AB - BACKGROUND/AIMS: The aim was to investigate the effect of blood inflow occlusion on lesion size and ultrasonographic findings during interstitial laser thermotherapy of normal liver. METHODOLOGY: Pigs were treated with or without hepatic inflow occlusion at a laser power of 3W or without inflow occlusion at 5 W (target temperature 43 degrees C). The thermotherapy system consisted of an Nd:YAG laser and a temperature feedback circuit. Ultrasonography was performed immediately after treatment. Lesion size was determined using light microscopy including immunohistochemistry with bromodeoxyuridine. RESULTS: Hyperechoic ultrasonographic changes were observed after treatment with inflow occlusion or when there was carbonization. If carbonization did not occur, unoccluded blood flow was associated with hypoechoic lesions. Following inflow occlusion, maximum lesion width 2 and 6 days after thermotherapy averaged 21.9 +/- 1.3 and 20.2 +/- 0.8 (means +/- SEM) mm, respectively. This was larger than the corresponding values of 10.8 +/- 0.8 and 11.1 +/- 2.0 observed after treatment without inflow occlusion at 3W (p < 0.01). Increase in laser power from 3 to 5W in experiments without inflow occlusion produced early carbonization and a slight increase in lesion size that did not match that produced by inflow occlusion. Ultrasound gave a correct prediction of necrosis size after treatment with inflow occlusion but overestimated the necrosis when inflow occlusion was not used. Ultrasound was furthermore unable to predict size of necrosis in individual experiments. CONCLUSION: Blood flow has a major influence on lesion size in interstitial laser thermotherapy of the liver and affects ultrasonographic images. Also, it appears that intraoperative ultrasonography cannot monitor lesion size with an accuracy that is sufficient for clinical use. PMID- 9356844 TI - Cystadenocarcinoma of the pancreas: characteristics and surgical strategy. AB - When a cystic lesion of the pancreas is encountered, the first priority is to differentiate whether it is an inflammatory or neoplastic cyst. Failure to recognize the true nature of a neoplastic cyst will lead to an incorrect treatment strategy. This was dramatically proven in one of our patients. Nevertheless, because of the slow growth of this type of tumor, its tendency not to infiltrate adjacent structures and the relatively low likelihood of metastasis, operative resection should be attempted in all patients, even in more advanced stages. PMID- 9356845 TI - Extrahepatic portal obstruction without hepatopetal pathway associated with congenital arterioportal fistula: a case report. AB - Extrahepatic portal obstruction is one of the causes of portal hypertension, in which well-developed hepatopetal pathways are commonly recognized. Herein an extremely rare case of extrahepatic portal obstruction without hepatopetal pathway, probably caused by arterioportal fistula, is reported. The patient was a normally matured 16-year-old girl admitted for further evaluation of jaundice, presenting with the clinical manifestations of the portal hypertension associated with hypersplenism and portosystemic venous shunt. Celiac angiography clearly demonstrated an intrahepatic arterial aneurysm fed by the right hepatic artery shunting to the superior mesenteric vein, and portography disclosed complete obstruction of the portal trunk with conspicuous hepatofugal pathway but no hepatopetal collateral veins. The exact mechanism of this phenomenon is not known and whether the extrahepatic portal obstruction was primary or secondary is still obscure. However, this is the first case report in the world literature describing extrahepatic portal obstruction with absence of hepatopetal pathway. PMID- 9356846 TI - Superior vena cava syndrome: the significance of endosonography in diagnosing enlarged mediastinal lymph nodes--a case report. AB - Superior vena cava syndrome is a rare, life-threatening clinical entity associated with occlusion of venous outflow from the head, neck and upper extremities. It is usually caused by an intrathoracic neoplasm, thrombosis, an aneurysm, or external compression. Benign diseases rarely cause this syndrome. Malignant neoplasms, including lymphoma, lung cancer and breast cancer frequently cause this syndrome. Herein, the case of a 63-year old patient who developed superior vena cava syndrome and dysphagia is reported. Endosonographic and CT investigation of the mediastinum confirmed enlarged lymph nodes exerting pressure on the superior vena cava and the esophagus, particularly at the level of the aortic arch. Cytologic examination of the lymph node specimen confirmed metastatic adenocarcinoma of the lung. The patient was treated by radiotherapy of the right lung and mediastinum. Patients with mediastinal tumors or enlarged lymph nodes frequently have dysphagic problems due to pressure on the esophagus. Endoscopy usually confirms a constriction of the lumen, but it cannot determine the cause. Endoscopic ultrasonography makes a precise differentiation between submucosal tumors and the causes of exterior compression possible. PMID- 9356847 TI - Congenital multiple arterioportal fistula: a case report treated with portocaval shunt. AB - A case of 48-year-old man, with diffuse arterioportal fistula (APF) considered congenital in origin, is described. The patient presented with recurring intractable variceal hemorrhage which was treated by endoscopic ligation or sclerotherapy. In this case, gastroduodenal artery and branches of mesenterial artery were diffusely involved, so these lesions could not be resected. Portocaval shunt between the superior mesenteric vein and the right iliac vein was successfully performed. Follow-up endoscopic examination one year later revealed no evidence of esophageal and gastric varices. To our knowledge, this is the second case in which a patient underwent portocaval shunt successfully for congenital APF. PMID- 9356848 TI - The role of acupuncture in the treatment of irritable bowel syndrome: a pilot study. AB - BACKGROUND/AIMS: The aim of this pilot study was to investigate the potential value of acupuncture in the treatment of irritable bowel syndrome (IBS). METHODOLOGY: The study was an open design study of 7 patients with established irritable bowel syndrome in which assessment was by a diary card. RESULTS: The results showed a significant improvement both in general well-being and in symptoms of bloating. CONCLUSIONS: Acupuncture seems to be effective in the treatment of irritable bowel syndrome and merits further study. PMID- 9356849 TI - Modified surgical techniques for the superlow anterior resection. AB - BACKGROUND/AIMS: Superlow anterior resection remains a technical challenge because it involves bowel resection and anastomosis in deep, limited pelvic space. Some modifications of the standard double stapling technique are thus needed to facilitate the procedure, making it safer, easier and more reliable. METHODOLOGY: A Roticulator is applied to make the first staple if the pelvic space is wide enough. A puncture hole is made manually in the closed rectal stump with the anvil tip under direct vision. If the pelvis is too narrow to apply a Roticulator, a purse-string suture is first placed before bowel resection, and is then tied securely around the anvil shaft to close the rectal stump. In either condition, the anvil and the attached sigmoid colon are pushed manually to evert the closed rectal stump. The stapled end-to-end anastomosis is then made outside the anus. RESULTS: From July 1994 to June 1996, 42 superlow anterior resections were performed using the modified techniques. There were only 2 cases of anastomotic leakage. The function results were acceptable. CONCLUSIONS: The modified techniques are safe, effective and easy to perform. Their major advantages are that the stapled anastomosis is made under direct vision and without the hindrance of pelvic tissue. PMID- 9356850 TI - Seasonal variation in exacerbations of ulcerative colitis. AB - BACKGROUND/AIMS: The aim of this study was to determine whether there is seasonal variability in exacerbations of ulcerative colitis. METHODOLOGY: The timing of ulcerative colitis relapses was retrospectively studied in a group of consecutive patients with quiescent ulcerative colitis. Ninety-four patients were followed-up at least every three months for a mean of 29.3 (range: 12-67) months. RESULTS: In total, 248 relapses of ulcerative colitis were observed with a mean number of 2.6 (range: 0-9) per patient. The timing of the relapses was characterized by a clear monthly and seasonal pattern (p < 0.001). In particular, the occurrence of relapses peaked during October and November (observed/expected (O/E): 30/20 in both months) and showed three troughs: during July and August, during December, and during February (O/E: 13/21, 7/21, 8/20, and 15/21, respectively). Moreover, the relapse rate was high during autumn and spring (O/E: 84/62 and 72/61, respectively) and low during summer and winter (O/E: 45/61 and 47/64, respectively). CONCLUSIONS: These data support the premise that there is seasonal variability in terms of relapses in ulcerative colitis patients and suggest that the role of seasonal triggering factors must be further investigated. PMID- 9356851 TI - Prophylactic perioperative treatment in a patient with colon carcinoma complicated by polycythemia vera. AB - A patient with colon carcinoma complicated by polycythemia vera (PV) who underwent a partial colectomy concomitant with prophylactic perioperative treatment resulting in successful outcome is herein described. Seven weeks after the cessation of the latest exacerbation of PV, a partial colectomy was performed. In order to prevent the development of disseminated intravascular coagulation and thrombotic complications, the following perioperative treatment was performed: administration of gabexate mesilate (2,000 mg/day), fresh frozen plasma (300 ml/day), heparin (5,000 IU/day) for 7 days and anti-thrombin-III for 4 days, and a potent antibiotic therapy for 12 days and graded elastic bandages around the bilateral lower extremities for 14 days. As a result, an uneventful postoperative course was achieved. The present case suggests that these treatments are useful in the perioperative management of PV patients. PMID- 9356852 TI - Fatal intestinal hemorrhage complicating ileal lymphoma after cyclosporine for unresponsive celiac disease. AB - BACKGROUND/AIMS: Unresponsive celiac disease may benefit from immunosuppressive therapy. Malignant small intestinal lymphoma is the most serious complication of celiac disease, also being noted as a complication of immunosuppressive therapy. The diagnosis of small intestinal lymphoma complicating celiac disease is notoriously difficult. Perforation is the most common complication of small intestinal lymphoma, frank hemorrhage being unusual. We report a case of massive, fatal hemorrhage from small intestinal lymphoma complicating unresponsive celiac disease treated with cyclosporine. The patient was presented with severe diarrhea and nutritional deterioration. Unresponsive celiac disease was diagnosed on the basis of clinical and histologic criteria with no response while on a gluten-free diet, corticotherapy and octreotide acetate injections. Cyclosporine therapy was advised. The patient had a remarkable clinical response. After 3 months from the start of the cyclosporine therapy, the patient returned with massive intestinal bleeding. The patient underwent emergency surgery diagnosing an enteropathy associated lymphoma. We conclude that cyclosporine therapy for unresponsive celiac disease should be considered in select, severely ill patients only after a full-thickness biopsy of the small intestinal wall to disclose a latent super imposed lymphoma, which course may be accelerated by immunosuppressive therapy. PMID- 9356853 TI - Modified intraparietal vagotomy in the treatment of perforated duodenal ulcer. AB - BACKGROUND/AIMS: The purpose of this study was to introduce modified intraparietal vagotomy as a safe procedure and a method of choice in the treatment of perforated duodenal ulcers. METHODOLOGY: Eighty-six patients with perforated duodenal ulcers underwent oversewing of the perforated ulcer and modified intraparietal selective vagotomy. The site of perforation was sewn over and an abdominal cavity lavage was performed. The posterior vagal nerve was resected, and a modified intraparietal anterior vagotomy was performed. During the postoperative period, after twenty days, six months and one year, respectively, we analyzed the following data: body weight, signs of gastroesophageal reflux, subjective discomfort, early postoperative complications, gastroduodenoscopic findings, basal acid output (BAO), and maximal acid output stimulated by pentagastrin (PAO). RESULTS: There was no mortality in our group, the post-operative morbidity was insignificant, and the duration of operation was shorter in comparison to other vagotomy methods. BAO and PAO values were similar to those in the literature when proximal selective vagotomy (PSV) was performed. There were no cases of duodenogastric or gastroesophageal reflux nor re-occurrence of ulcer disease, as confirmed by gastroduodenoscopy. According to the modified Visick's criteria, 94% of the patients followed-up were classified as group 1. CONCLUSION: We consider the modified intraparietal vagotomy to be the method of choice in the treatment of perforated duodenal ulcers because of the simple surgical technique involved, the shorter duration of surgery, and the avoidance of standard PSV complications. The surgery can be performed even by a less experienced surgeon, independently of the patient's age and condition. This modification is suitable for laparoscopic surgery. PMID- 9356854 TI - Duodenal obstruction by gallstones (Bouveret's syndrome). Presentation of a new case and literature review. AB - Bouveret's syndrome is a rare entity consisting in a duodenal obstruction due to the passage of gallstones from the gallbladder to the duodenum through a cholecystoduodenal fistula. Approximately 225 cases are reported in the literature. It is most common in old women with a previous history of biliary tract disease. The clinical picture is nonspecific and pre-operative diagnosis is not easy. Oral endoscopy is the main diagnostic procedure and sometimes, a therapeutic option, too. Surgery is the elective treatment specially when endoscopy is unsuccessful. We report a new case of this syndrome successfully treated by surgery, and an extensive review of the literature concerning this issue, focusing mainly on the clinical findings, diagnosis, therapeutic procedures and results. We conclude that Bouveret's syndrome is rare but more frequent in older females with previous biliary disease, better diagnosed by pyloric obstruction syndrome, plain abdominal x-ray, ultrasonography, contrast gastric study and/or gastroscopy (confirming and best procedure). When conservative endoscopic procedure fails, surgical treatment must be carried out, thus obtaining good results. PMID- 9356855 TI - Orthotopic liver transplantation for alcoholic liver disease: rates of survival, complications and relapse. AB - BACKGROUND/AIMS: Liver transplantation for alcoholic end-stage liver disease remains controversial at many transplant centers. The aim of the present study was to evaluate the outcome of patients with alcoholic liver disease who underwent liver transplantation at Centro Trapianti, Policlinico S. Orsola, Bologna. METHODOLOGY: We describe the outcomes of 18 alcoholic patients with end stage liver disease who received orthotopic liver transplants at our center from April, 1986 to February, 1996. The data obtained was compared with that of 114 patients with virus-related cirrhosis selected as transplant controls. An absolute period of abstinence from alcohol consumption for at least six months was required. RESULTS: Regarding the actuarial survival rate and non-fatal post transplant complications, no significant differences were noted in comparing the non-alcoholic with the alcoholic recipients, except for a higher incidence of Cytomegalovirus infection in the alcoholic group followed-up for more than four months. The alcoholic relapse rate was 27.2%, but only one patient returned to harmful drinking. Seventy-three percent of subjects who were followed-up for at least six months were occupied in gainful employment. Alcoholic relapse did not affect employment status. CONCLUSION: This data demonstrates that liver transplantation for selected patients with end-stage alcohol-related cirrhosis achieves good results in survival, complications and employment status, and it appears difficult to defend an inflexible claim to have demonstrated an absolute long-term abstinence before transplantation when a severe deterioration of liver function is present. PMID- 9356856 TI - Abstinence-induced oxidative stress in moderate drinkers is improved by bionormalizer. AB - BACKGROUND/AIMS: The aim of this investigation was to study the oxidative phenomena which take place in the early recovery phase after alcohol withdrawal. Furthermore, the effects of a novel natural antioxidant, Bionormalizer (BN), in such a clinical setting was studied. METHODOLOGY: Forty-six alcoholics with moderate drinking habits (daily ethanol intake: > 80g to < 120g) were enrolled in the study, divided into two groups and given either a placebo or 9g of BN by mouth every night for one week. The patients agreed to stop drinking alcohol, and daily blood sampling was obtained for routine tests and to check plasma and erythrocyte levels of MDA, SOD, GPX and the hydroperoxide level. The groups were comparable in terms of initial biochemical parameters. RESULTS: BN prevented the early increase of plasma TBARS observed in the placebo group, enabling a near-to normal level of plasma and erythrocyte MDA by the fourth day. BN also prevented the significant drop of erythrocyte GPX and the transient decrease of plasma SOD observed in the placebo group. Despite alcohol withdrawal, plasma lipid hydroperoxide remained significantly elevated in the placebo group, but this phenomenon was rapidly improved by BN. CONCLUSIONS: To a significant extent, BN is able to prevent the free radical-mediated lipoperoxidative changes that occur soon after alcohol withdrawal, while fastening the recovery mechanisms. PMID- 9356857 TI - Physician's working diagnosis compared to the Euricterus Real Life Data Diagnostic Tool Trial in three jaundice databases: Euricterus Dutch, independent prospective and independent retrospective. AB - BACKGROUND/AIMS: In the European Union Euricterus Project on (sub)Icterus proforma, the history and physical examination items were to be used for the physician's working diagnosis (PWD) and 'among others, for the development of the real life data electronic diagnostic tool, Trial. Trial delivers diagnosis probabilities based on Bayes' Theorem (B), completed by Trial Algorithm (TA). We wanted to compare the diagnostic accuracies (PWD and Trial probabilities as a percentage of the final diagnosis (FD) in a patient population) in 3 Dutch databases. METHODOLOGY: The inclusion criteria for both Euricterus and Trial were age > or = 16 and bilirubin > or = 20 mmol/l. Euricterus data gathering took place at the bedside on a proforma with (among other questions) 79 questions on history and physical examination as well as the diagnosis levels for the PWD (1 alternative possible) and FD (17 disease categories, dc). Trial was developed on the data of 7,104 Euricterus patients and its data-entry Demo has the same questions. It calculates the probability of each diagnosis of the 17 dc as a percentage, as each significant finding is encountered (BO, Bayesian Overall). It can simultaneously calculate the resemblance of the patient's signs and symptoms to each disease concomitantly (BV, Bayesian Vertical), and to any subset of a disease. Any probability is further tested for compatibility using TA, a subset of BV, delivering TA-PWD, TA-BO and TA-BV. The Trial test patients came from 3 databases: a Euricterus Dutch Patients Random Sample EDRS (n = 184, internal database) and 2 independent databases: prospective P (n = 80) and retrospective R (n = 152), totalling 416 patients. RESULTS: The accuracies of PWD and Trial showed no differences between the databases, and the results are therefore pooled (n = 416). With testing on the highest probability found, the PWD accuracy was 78%, TA-PWD 81%, TA-BO 74% and TA-BV 72%. The true FD's were mentioned (at any probability) in the PWD in 86%, TA-PWD in 92%, TA-BO in 94% and TA-BV in 91% of the patients. Testing only patients whose FD was "certain" or whose data were without omissions did not improve accuracy. Testing on probability > 95% improved BO and BV accuracy, but not TA-BO or TA-BV. CONCLUSIONS: The Physician's Working Diagnosis accuracy was approximately 80% and did not greatly improve after TA. The Trial TA-BO and TA-BV accuracies were only slightly less than the PWD. For well-trained physicians, Trial strengthens the physician's judgment, and for those less trained (or those to be trained), it delivers a (sub)icterus diagnostic disease probability at nearly consultant level. PMID- 9356858 TI - Assessment of liver cirrhosis severity in 1015 patients of the Euricterus database with Campbell-Child, Pugh-Child and with ascites and ascites-nutritional state (ANS) related classifications. Euricterus Project Management Group. AB - BACKGROUND/AIMS: The assessment of disease stage in cirrhosis is important for the individual patient (prognosis, timing and risk for requiring surgical intervention) and also for population comparisons and trials. There are several established methods, and we have aimed at comparison of the methods within a large cirrhosis population. METHODOLOGY: In the European Union Euricterus database, there are 1015 patients with a "certain" diagnosis of cirrhosis, each of whom in one session had a protocol work-up of history, physical examination and all laboratory investigations needed for this study. The Child-Turcotte (CT), Campbell-Child (C) and Pugh-Child (P) classifications, as well as ascites/no ascites, ascites 1, 2, 3 (no, therapy responsive, nonresponsive) and ascites/nutritional state (ANS, 1-9) scores were used. CT and C have the same 5 variables, P has prothrombin time instead of nutritional state. CT, C and P variables score 1-3 each. C and P furthermore have variable range scores of 5-15. CT, C and P have classes A-C. The variables used were ascites, nutritional state, encephalopathy, bilirubin, albumin and prothrombin time. RESULTS: Only 53 patients (5%) fit within the CT criteria. C and P variable range scores (5-15) correlated strongly (r = 0.84). Cross-over calculation showed slightly different results in the P and C choice of variables, while the variable ranges (1-3) did not matter. Different selection of score ranges for the A-C classes in C and P resulted in 69% class C in P (35% in C) and 3% A in P (19% in C). The patients with ascites (70%) had worse bilirubin, albumin, nutritional states and C and P 5 15 scores (p < 0.0001). Patients with ascites 3 had all variables and also C, P 5 15 scores worse than those with ascites 2 (p < 0.02). ANS scoring showed wasting in 33% of the patients without ascites (ANS 3), 50% of the patients with ascites 2 (ANS 6) and 60% with ascites 3 (ANS 9) (p < 0.0003), and C and P scores were higher in the 3 ANS scores with wasting. CONCLUSIONS: Campbell and Pugh 5-15 scores correlated closely and can be used interachangeably. As C does not contain the more elaborate prothrombin time determination, it probably can be used anywhere in the world. Ascites (degree) and Ascites/Nutritional State (ANS) scoring only use history and physical examination and are, or remain, although less refined, clinically relevant. PMID- 9356859 TI - Left trisegmentectomy for hepatocellular carcinoma with tumor thrombi extending to the third branches in the remnant liver. AB - BACKGROUND/AIMS: A successful left trisegmentectomy and tumor thrombectomy for hepatocellular carcinoma (HCC) of the right third branches in the remnant liver is described. METHODOLOGY: In this case, the hepatocellular carcinoma originated in the internal segment of an HBs- and HBe-Ag positive cirrhotic liver, involved the portal vein of the umbilical portion across the portal trunk to the contralateral third branches, and had many metastatic nodules in the anterior segment of the liver. To remove tumor thrombi by directly visualizing the lumen of the third branches of the portal vein, the portal trunk to the anterior branch of the portal vein was completely isolated by transecting the hepatic parenchyma along the intersegmental plane through a hilar approach. Tumor thrombi were removed by incising the portal trunk toward the anterior branch of the portal vein. Portal flow to the remnant liver was restored preceding liver resection to preserve hepatic function. After division of the anterior Glissonean code, the left trisegment and caudate lobes of the liver were resected. RESULTS: The patient was discharged 10 weeks after surgery and remained well for the first six months thereafter. Recurrent tumors, however, appeared in the remnant liver 7 months after surgery. CONCLUSIONS: This procedure may be applicable in cases of HCC with portal tumor thrombi extending into the third branches in the remnant liver. PMID- 9356860 TI - Adverse effect of perioperative blood transfusions on survival after hepatic resection for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Experimental and clinical studies have found a relationship between blood transfusion and altered immune function. We estimated the risk of transfusions for shorter survival on patients with hepatocellular carcinoma who underwent hepatic resection. METHODOLOGY: The impact of perioperative blood transfusions on 235 patients with hepatocellular carcinoma who had resections from January 1981 to December 1988 was retrospectively examined. All patients underwent hepatic resection and received no additional chemotherapy. RESULTS: Using the Cox proportional hazard model, the number of perioperative blood transfusions was found to be a significant prognostic factor for patient outcome (p = 0.03). Overall, patients who received less than 12 transfused units had a significantly better 5-year survival rate than those who received more than 13 transfused units (46.3% vs. 24.5%, p < 0.001). This result was also seen when the patients were subdivided by stage: 5-year survival in the early stage group (57.2% vs. 35.5%, p < 0.01) and in the advanced stage group (30.0% vs. 18.2%, p < 0.05). The number of perioperative blood transfusions also influenced the survival of patients who underwent a curative resection (66.2% vs. 38.5%, p < 0.01), but did not affect the survival of those who received a non-curative resection (7.9% vs. 7.4%). CONCLUSION: This study suggests that the number of perioperative blood transfusions is a significant prognostic factor in patients with hepatocellular carcinoma who undergo hepatic resection. PMID- 9356861 TI - Comparison between thoracoabdominal and abdominal approaches in occurrence of pleural effusion after liver cancer surgery. AB - BACKGROUND/AIMS: Pleural effusion is a complication occasionally encountered in hepatic surgery. The production of pleural effusion was compared between thoracoabdominal and abdominal approach for hepatic surgery of hepatocellular carcinoma. METHODOLOGY: All the 98 patients undergoing liver resection for hepatocellular carcinoma at the National Cancer Center Hospital from May 1992 to March 1994 were included into the study, of those 70 were by the thoracoabdominal and 28 by the standard abdominal approach. Comparisons were made in regard to the rate of pleural effusion, the rate of postoperative thoracentesis, the number of postoperative thoracentesis procedures per patient, the total volume of pleural effusion obtained by thoracentesis per patient, and the duration of pleural effusion. RESULTS: Forty-three percent of patients treated with the abdominal approach in contrast to 73% of patients treated with the thoracoabdominal approach developed pleural effusion (p < 0.01). Seven percent of the patients treated with the abdominal approach in contrast to 17% of the patients treated by the thoraco-abdominal approach required thoracentesis. The number of thoracentesis required for the abdominal approach was 1, for the thoracoabdominal approach was 3 (p < 0.02). The bile leakage rate was 17% for thoracoabdominal versus 33% for abdominal approach. CONCLUSIONS: Due to frequent pleural effusion, the thoracoabdominal approach should not be used for every liver operation. However, when treatment to the neck of the right hepatic vein is necessary, the thoracoabdominal approach might be recommended because of the easy access to the operating field and the reduced rate of bile leakage. PMID- 9356862 TI - Hepatocellular carcinoma and hepatic cirrhosis in Mexico: a 25 year necroscopy review. AB - BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is a common form of cancer which is found throughout the world. In recent years, the rates of HCC seem to have increased in European and North American countries. Herein, a retrospective review of necroscopy records over a 25-year period was undertaken in order to determine the incidence of HCC in a Mexican population. METHODOLOGY: A retrospective review of necroscopy records was performed to determine the incidence of HCC and then to determine the presence or absence of associated cirrhosis in these cases. The tumor/cirrhosis relationship was determined by computing the TC/T and TC/C indexes. RESULTS: Of 12556 autopsies studied, 73 cases of histologically proven HCC were reported, representing a total necropsy carcinoma incidence of 0.59%. Fifty-five cases were associated with cirrhosis (0.43%), and 18 were not (0.14%). HCC was two times more common in males (67%) than in females (33%), with a ratio of 2:1. During this period, the necropsy incidence of HCC rose steadily to twice its original level (1965-69 incidence 0.35%; 1985-89 incidence 0.69%). The necropsy incidence of cirrhosis was 4% (329 males, 185 females). The overall TC/T index was 75% (87% for males and 50% for females). The overall TC/C index was 10.7% (13% for males and 6.4% for females). CONCLUSIONS: There was a two-fold increase in the incidence of HCC in the Mexican population studied over a 25-year period. HCC was associated with cirrhosis in the majority of cases. HCC was two times more common in males than in females in patients with cirrhosis; in patients without cirrhosis, the ratio was 1:1. The incidence of cirrhosis was 4%, which remained unchanged with the passage of time. PMID- 9356863 TI - Dual infection with hepatitis C and B viruses: clinical and histological study in Saudi patients. AB - BACKGROUND/AIMS: The purpose of this study was to assess the clinical and histological significance of dual infection with Hepatitis C virus and Hepatitis B virus. METHODOLOGY: The clinical presentation and histologic findings of 20 patients with HCV and HBV were reviewed and compared to 33 patients with only HCV, as controls. None of the patients were treated and none had anti-HDV or HIV 1/HIV2. RESULTS: The patients with dual HCV and HBV infection had more decompensated liver disease. Most of these patients were classified in the Child Pugh group C as compared to the controls (36.8% vs 0%, p < 0.01). Liver cirrhosis was more common in patients with HCV and HBV infection than it was in the controls infected with HCV only (95% vs 48.5%, p < 0.01). Similarly, hepatocellular carcinoma was more common in patients with dual HCV and HBV infection than it was in the controls (63% vs 15%, p < 0.01). CONCLUSION: Liver disease seems to be more severe in patients with dual HCV and HBV infection than in patients with HCV infection only, both clinically and histologically. PMID- 9356864 TI - Characteristic differences of hepatocellular carcinoma in Japan, with special reference to liver weight at autopsy. AB - BACKGROUND/AIMS: Liver weight at autopsy in patients with hepatocellular carcinoma (HCC) has been found to differ by geographic region. The clinicopathological characteristics of HCC in relation to liver weight at autopsy have not been reported for Japanese cases. METHODOLOGY: From 1980 to 1992, a total of 107 consecutive HCC autopsy cases were studied. Seven cases were excluded (5 surgically resected cases and 2 cases of occult-type HCC (Berman) which were diagnosed at autopsy for the first time). The 100 cases of HCC were divided into 4 groups according to liver weight at autopsy: less than 999 g (group A, n = 15, 15%); 1000-1999 g (group B, n = 50, 50%); 2,000-2,999 g (group C, n = 20, 20%); and more than 3,000 g (group D, n = 15, 15%). Various clinical and pathological findings, including age, sex, hepatitis B surface antigen (HBsAg) status, history of blood transfusions, treatment, cause of death, underlying liver disease, survival after diagnosis, tumor size, macroscopic findings, microscopic findings, splenic weight and liver weight were studied. RESULTS: Significant differences between groups were found in sex (p < 0.01), underlying liver disease (p < 0.001), tumor size (p < 0.0001), microscopic findings (p < 0.001) and survival (p < 0.01). The lowest liver weight group (group A) had a significantly longer mean survival rate than the other three groups (p < 0.001). CONCLUSIONS: These results showed that 15% of HCC occurred in large size livers (more than 3,000 g), 15% occurred in small size livers (less than 999 g) and 70% occurred in livers between 1,000 g and 2,999 g, and that tumor size (p < 0.0001) and microscopic findings (p < 0.001) were the major determinants of liver weight in HCC patients. The total liver volume reflected by liver weight in HCC cases may be valuable for the assessment of the clinicopathological features of HCC and its prognosis. PMID- 9356865 TI - False-positive endotoxemia derives from gauze glucan after hepatectomy for hepatocellular carcinoma with cirrhosis. AB - BACKGROUND/AIMS: The purpose of this study was to evaluate the occurrence of false-positive endotoxemia after hepatectomy for hepatocellular carcinoma in patients with cirrhosis. The chromogenic conventional limulus test (Toxicolor test) revealed transiently increased blood endotoxin levels after hepatic resection for hepatocellular carcinoma in patients with liver cirrhosis. However, clinical signs of endotoxemia were not observed in all of the cases. METHODOLOGY: Pre- and postoperative changes of blood endotoxin levels and beta-glucan were measured in 20 patients with liver cirrhosis who underwent hepatic resection for hepatocellular carcinoma, using the Toxicolor test, the Endospecy test (which specifically reacts with endotoxin) and the Gluspecy test (which specifically reacts with beta-glucan). The changes in endotoxin levels and beta-glucan in ascitic fluid during surgery were also studied. RESULTS: The Toxicolor test revealed transiently increased blood endotoxin levels, but Endospecy test results were not elevated. The changes in the Gluspecy test were almost the same as in the Toxicolor test. During surgery, beta-glucan levels in the ascitic fluid increased remarkably due to their release from the surgical gauze. Digestion studies using 1-3-beta-D-glucanase on specimens which showed high Toxicolor and Gluspecy test values resulted in a lack of response for both tests. CONCLUSION: The cause of false-positive endotoxemia was determined to be caused by beta glucan, which was released from the surgical gauze. PMID- 9356866 TI - Effect of misoprostol on the course of viral hepatitis B. AB - BACKGROUND/AIMS: Some prostaglandins revealed hepatoprotective effects, that was confirmed mostly in experimental liver injury. The aim of the study was to determine the effects of misoprostol on the course of viral hepatitis B and associated gastric mucosal injury. METHODOLOGY: Fifty two male patients with viral hepatitis B were assigned at random to receive either misoprostol (800 micrograms/day), silymarin (210 mg/day) or no drug treatment at all (control group). Biochemical indices of liver injury were measured once a week, and HBsAg clearance was analysed 6 months later. Moreover serum levels of endogenous prostaglandins (PGE2, PGI2) and stomach mucosal injury (scored endoscopically) were evaluated before and after treatment. RESULTS: Decrease of serum concentration of bilirubin, activities of transaminases and alkaline phosphatase as well as hepatomegaly reduction were significantly faster in misoprostol treated patients, that resulted in significantly shorter time of hospitalization. The best effect of misoprostol treatment on liver injury was observed in patients suffering from a severe course of the disease. HBV elimination, evaluated 6 months after the disease onset, was similar in both groups. Misoprostol treatment had a significant effect on the improvement of stomach mucosal injury. Serum concentrations of main endogenous prostaglandins produced by the liver (PGE2 and PGI2), were not affected by exogenous administration of misoprostol. There were no side effects related to misoprostol or silymarin treatment. CONCLUSIONS: Our data demonstrates the beneficial effect of misoprostol treatment in patients with viral hepatitis B. Faster convalescence related to normalization of biochemical indices of liver injury and healing of associated stomach mucosal lesions was also observed. PMID- 9356867 TI - Auxiliary heterotopic partial liver transplantation in pigs with acute ischemic liver failure. AB - BACKGROUND/AIMS: Fulminant hepatic failure (FHF) is usually fatal without liver transplantation. Auxiliary heterotopic partial liver transplantation (AHPLT) may offer advantages over orthotopic liver transplantation (OLT) or any other heterotopic procedure for the treatment of patients with fulminant liver failure. We studied AHPLT in a severe acute hepatic failure model in pigs. METHODOLOGY: Group A (control: n = 5) underwent portal vein and hepatic artery ligation and side-to-side portocaval shunting. Group B (AHPLT: n = 15) underwent host portal vein and hepatic artery ligation and AHPLT. RESULTS: All of the pigs in group A died within 48 hours from massive liver necrosis. Ten of the 15 pigs (67%) in group B had well-functioning grafts. Five of these ten died between 8 and 17 days postoperatively due to various complications. The remaining five survived for sixty days postoperatively in healthy condition. At the time of sacrifice, four of these five had well-functioning grafts weighing 739 +/- 52 g (mean +/- SEM) and regenerated, but still atrophied, host livers weighing 262 +/- 23 g (p < 0.0002). On the other hand, the one remaining pig had an atrophied graft weighing 310 g and a well-regenerated host liver weighing 470 g, probably due to a late, poorly functioning graft associated with severe rejection. CONCLUSION: AHPLT may result in survival despite host hepatic failure, and the host liver may recover within two months, despite total interruption of blood inflow. PMID- 9356868 TI - Hepatic resection for icteric type hepatocellular carcinoma. AB - Icteric-type hepatocellular carcinoma, which initially presents as jaundice, is known to be rare. Furthermore, the number of such cases that undergo hepatic resection is also very small. The purpose of this study was to clarify the characteristics of icteric type hepatocellular carcinoma and discuss the efficacy of hepatic resection in this condition. Herein, we present five cases of icteric type hepatocellular carcinoma which were among a study of 438 patients who underwent hepatic resection. Most of these cases were in the advanced stages, and a high incidence of early death was recognized. However, two patients are doing well, without further recurrence (8 years and 7 months, and 13 months, respectively). It is important to consider icteric type hepatocellular carcinoma whenever a patient has a potential risk for hepatocellular carcinoma. In addition, it is also important to understand that in diagnosing icteric type HCC, sometimes neither choledocholithiasis nor cholangiocellular carcinoma can be clearly ruled out. Extensive examinations of the biliary tract, including percutaneous transhepatic cholangiography as well as endoscopic retrograde cholangiography are indicated for such patients when they exhibit either temporary cholangitis or jaundice, as well as when there is biliary dilatation within the liver. Furthermore, hepatic resection is also considered to be a viable alternative for such cases. PMID- 9356869 TI - Arterial ketone body ratio during hepatectomy. AB - The arterial ketone body ratio (AKBR) has been proposed as an accurate indicator of liver mitochondrial redox potential. However, the efficacy of the AKBR as a biochemical marker has been recently called into question. To resolve this issue, we studied the effect of temporary vascular occlusion on the AKBR during hepatectomy. Twenty patients undergoing hepatectomy were divided into two groups: those with hepatocellular carcinoma with a history of hepatic cirrhosis (n = 10; cirrhotic group) and those with liver disease without cirrhosis (n = 10; non cirrhotic group). To minimize blood loss during hepatectomy, temporary vascular occlusion was applied using the Pringle maneuver. Acetoacetate and beta hydroxybutyrate concentrations in the arterial blood and the AKBR were determined before and after vascular occlusion. In 25% of the two groups combined, the AKBR increased following normothermic ischemia, as compared with the levels prior to clamping; in 20% of cases in the cirrhotic group, it increased immediately following reperfusion, as compared with the levels prior to clamping. Changes in the AKBR during hepatectomy did not correlate with preoperative hepatocellular function. An AKBR of less than 0.7 prior to clamping which persisted during surgery was not a consistent risk factor for postoperative complications. The AKBR was not a useful predictor of liver viability in partial liver resection with temporary vascular occlusion. PMID- 9356870 TI - The relationship between DNA content, clinicopathology and prognosis in enteropancreatic carcinoids. AB - BACKGROUND/AIMS: The clinical behavior of carcinoid tumors is sometimes difficult to determine from their histological appearance. The aim of the present study was to evaluate whether the DNA distribution pattern seen in carcinoid tumors can be correlated with histopathological parameters and the patient's clinical course and prognosis. METHODOLOGY: The paraffin-embedded material of 44 enteropancreatic tumors underwent deparaffinization, was rehydrated, and mechanically and enzymatically processed into a single cell solution. For evaluation of the DNA histogram, analysis was performed with the help of an automatic single cell cytophotometric study. RESULTS: A correlation was seen between the DNA content and tumor stage (p = 0.01), radicalness of surgery (p = 0.03), tumor localization (p = 0.02) and patient's age (p = 0.05). In univariate analysis, patient's age (p = 0.04), tumor localization (p = 0.03), tumor stage (p = 0.0001), radicalness of surgery (p = 0.0006) and DNA content (p = 0.01) influenced prognosis. In multivariate analysis including these parameters, only tumor stage had an independent influence on prognosis. CONCLUSION: The clinical relevance of DNA measurement in carcinoid tumors is still unknown. PMID- 9356871 TI - A comparison of the complication rate for three pancreaticojejunostomy techniques. AB - BACKGROUND/AIMS: A variety of techniques for pancreaticojejunostomy have been employed with introduction of safer anastomosing methods in our department as an extended excision of the pancreas was performed. In this study, complications associated with the various techniques of pancreaticojejunostomy were investigated in order to find a safer anastomosing method. METHODOLOGY: The extent of excision and complication rate of three types of pancreaticojejunostomy (impaction method, subserosal anastomosis and non-division procedure) were investigated in 64 patients who underwent pancreatoduodenectomy for diseases of the pancreas and biliary system. The end-to-side anastomosis of the pancreas and jejunum without division of the jejunoserosal tunica muscularis (non-division method) is an anastomotic method with a low risk of suture insufficiency. RESULTS: The incidence of secondary complications in the patients with suture insufficiency was higher than that in the patients with leakage of pancreatic juice. Once leakage of pancreatic juice and suture insufficiency occurred, the incidence of secondary complications was high after extended excision. CONCLUSIONS: From these results, the non-division method was found to be a safe anastomosing method. After an extended excision is performed, intraperitoneal drainage is necessary. PMID- 9356873 TI - Predictive factors for long-term survival in patients with pancreatic carcinoma. AB - BACKGROUND/AIMS: Although numerous clinico-pathological parameters have been demonstrated to predict the prognosis in patients with pancreatic carcinoma after surgical resection, the factor most significant for their post-operative survival is still controversial. Herein, the authors have performed histopathological studies on patients with pancreatic adenocarcinoma and reviewed their clinical records to detect the most significant factor influencing their long term survival. METHODOLOGY: We reviewed clinical records and histological findings retrospectively. Then we performed univariate and multivariate analyses to find a correlation between the factors and the survival of the patients. RESULTS: The overall postoperative morbidity and mortality rates were 30.0% and 5.6%, respectively. The overall 5-year survival was 9.0%. Univariate analysis resulted in the detection of 14 factors which correlated with patient prognosis. Multivariate analysis resulted in the detection of histologic differentiation as an independent predictor for longterm survival. CONCLUSION: Histological differentiation is recommended as the most reliable predictor for the prognosis of patients with pancreatic carcinoma after surgical removal. PMID- 9356872 TI - Ras gene point mutations in gallbladder lesions associated with anomalous connection of pancreatobiliary ducts. AB - BACKGROUND/AIMS: The present study was undertaken to investigate possible changes in the K-ras oncogene in patients with gallbladder lesions (carcinoma, adenoma or hyperplasia) in relation to the presence or absence of an anomalous connection of pancreatobiliary ducts (ACPBD). METHODOLOGY: Gallbladder specimens were obtained from 44 patients with lesions that were either with or without ACPBD, and DNA samples were analyzed using PCR-SSCP. Point mutations in codons 12, 13 and 61 were analyzed by direct sequencing methods with oligonucleotide primers. RESULTS: The K-ras codon 12 was detected in 83.3% (5/6) of carcinomas the one adenoma tested and in 35.7% (5/14) of hyperplastic lesions with ACPBD, as opposed to only 36.4% (4/11) of carcinomas without ACPBD. The one case of gallbladder adenoma and 11 cases of normal gallbladder without ACPBD studied demonstrated no point mutations in the K-ras oncogene. CONCLUSIONS: Alteration of the K-ras oncogene appears to be involved in the early stages of gallbladder carcinogenesis when in association with ACPBD. The results further suggest that hyperplasia in cases with ACPBD may be a significant pre-cancerous lesion. PMID- 9356874 TI - Intraoperative radiotherapy for pancreatic carcinoma with hepatic or peritoneal metastases. AB - BACKGROUND/AIMS: The purpose of this study was to determine the efficacy of intraoperative radiotherapy (IORT) for unresectable pancreatic carcinoma associated with hepatic or peritoneal metastasis. METHODOLOGY: Between 1991 and 1994, 53 patients with pancreatic carcinoma associated with hepatic or peritoneal metastasis underwent surgery. Twenty-four of these patients received IORT, while 29 received no radiation therapy. The efficacy of IORT on the postoperative survival and pain relief for these patients was retrospectively analyzed. RESULTS: Postoperative survival was lowest in the subgroup of patients (n = 18) with both hepatic and peritoneal metastases, and this group did not benefit from IORT (IORT, n = 6; no IORT, n = 12) in terms of survival. Similarly, there was no significant difference in the survival rates between patients undergoing IORT (n = 10) and patients without IORT (n = 11) in the subgroup of patients with hepatic metastasis but without peritoneal metastasis. However, patients with peritoneal metastasis but without hepatic metastasis benefited significantly from IORT (IORT, n = 8; no IORT, n = 6) (p < 0.05). Pain relief following IORT was observed in 9 out of 10 patients who had experienced pain prior to surgery. CONCLUSION: Pancreatic carcinoma associated with peritoneal metastasis but without hepatic metastasis can be palliated by IORT. In addition, pain palliation in patients who require gastrointestinal or biliary drainage can also be achieved by IORT. PMID- 9356875 TI - Our approach to the surgical treatment of chronic pancreatitis. AB - BACKGROUND/AIMS: Optimal surgical management of the patients suffering of chronic pancreatitis still remains a controversial problem. The base for the evaluation of patients is for us the Marseille-Roma classification from 1988 with 4 fundamental types of the disease. Surgical therapy is usually indicated in patients with intractable or continuous pain, stenosis of the common bile duct and duodenal stenosis. The source of the above mentioned problems is related to the head of the pancreas and we prefer as the procedure of choice duodenum sparing resection of the pancreatic head. METHODOLOGY: Seventy patients were treated by our team in the years from 1985 to 1995 surgically and 55 of these patients underwent resection of the head of the pancreas. RESULTS: Eighteen patients were treated by hemipancreatoduodenectomy and 37 patients by the duodenum sparing resection of the pancreatic head. CONCLUSION: Left sided resection and drainage procedures appear to be much less effective from the point of view of long term follow-up. PMID- 9356876 TI - Bile duct carcinoma without jaundice: clues to early diagnosis. AB - BACKGROUND/AIMS: Most bile duct carcinomas are diagnosed at an advanced stage, after the appearance of jaundice. The features of bile duct carcinomas without jaundice were analyzed with the aim of allowing early diagnosis in such cases. METHODOLOGY: Clinicopathological features, images and surgical outcomes were compared between 18 non-jaundiced and 85 jaundiced patients with extrahepatic bile duct carcinoma. RESULTS: Among the non-jaundiced patients, 13 were symptomatic. Abnormalities on hepatic function and tumor marker tests were seen in 56% and 44%, respectively. In all 18 cases, ultrasonography demonstrated biliary abnormalities including masses (9 patients) and strictures (5 patients). The diagnosis was confirmed histologically by transpapillary bile duct biopsy in eight of 10 non-jaundiced patients. The non-jaundiced patients (83%) had a higher rate of resectability than jaundiced patients (58%). Pathological findings of resected specimens showed no significant differences between the two groups. The non-jaundiced group had a significantly higher survival rate than the jaundiced group: 50% vs. 22% at 5 years. CONCLUSION: For early diagnosis of bile duct carcinomas not associated with jaundice, detailed ultrasonographic examination is useful, and subtle changes indicate a need for direct cholangiography. Non jaundiced cases have the potential for curative resection. PMID- 9356877 TI - Metastatic liver disease--a review. AB - A common site of metastasis due to various forms of primary malignancies, is the liver. At present, increased screening of patients at risk has allowed early detection of the disease. Consequently, surgeons have started to perform hepatic resection for metastatic cancer, with the help of new or improved techniques with an enlarged spectrum of indications. New therapeutic modalities have also become available with the use of anti-cancer drugs, via new delivery devices, prior or after liver resection. Locoregional transarterial immuno-chemotherapy and isolated liver perfusion belong to this form of therapy. Although prognosis has improved impressively and low mortality and morbidity have emerged through improved surgical procedures, many patients still succumb to the disease due to locoregional recurrence of metastatic liver disease. PMID- 9356878 TI - Adenocarcinoma in the middle third of the stomach--an evaluation for the prognostic significance of clinicopathological features. AB - BACKGROUND/AIMS: The prognosis of patients with gastric adenocarcinoma varies with the location of the tumor. Adenocarcinoma in the middle third of the stomach has been claimed to have a better outcome than those in other locations. However, there is still very limited information specifically regarding the prognostic factors which influence the survival time of patients with adenocarcinoma in the middle third of the stomach. This retrospective study was designed with the aim to evaluate and uncover the possible significant clinicopathological parameters for adenocarcinoma in the middle third of the stomach. METHODOLOGY: Between 1986 and 1992, 363 patients underwent gastric resection for primary gastric adenocarcinoma at this hospital. Fifty-two (14.3%) of these patients were included in this study and they all met the following criteria: 1) tumor primarily located in the middle third of the stomach without distant metastases or peritoneal seeding, 2) undergoing curative resection and 3) undergoing R2 nodal dissection, at least. The clinicopathological findings were obtained by detailed review of the medical records and the histologic slides. All surviving patients were also contacted and their current conditions were recorded. RESULTS: The overall 5-year survival rate (Kaplan-Meier method) was 42.5%. In univariate survival analysis by Kaplan-Meier method and long-rank test, serosal invasion (p < 0.01), lymph node metastasis (p < 0.01) and lymphatic involvement (p < 0.01) had an individual prognostic significance. When a multivariate analysis using Cox proportional hazards regression was performed, serosal invasion (P < 0.01) and lymphatic involvement (p < 0.05) appeared as the only two independent prognostic factors regarding long-term survival. When these 52 patients were categorized into patients with early gastric cancer (n = 10) and patients with advanced gastric cancer (n = 42), there was a significant difference (p < 0.01) between the survival rates (90.0% vs. 29.1%). When these tumors were further categorized into early gastric cancer (n = 10), early simulating advanced gastric cancer (n = 14) and Borrmann type advanced gastric cancer (n = 28), there were significant differences (P < 0.01 and P < 0.01, respectively) in 5-year overall survival rates between early gastric cancer (90.0%) and Borrmann type advanced gastric cancer (18.9%), also between early simulating advanced gastric cancer (52.5%) and Borrmann type advanced gastric cancer (18.9%). UICC stage also had significant influence (P < 0.01) on the survival rates. CONCLUSIONS: Serosal invasion and lymphatic involvement are the significant, independent prognostic factors in predicting the survival rate of patients with adenocarcinoma in the middle third of the stomach. Since more advanced stage tumors usually carry a poorer prognosis, early detection is of extreme importance for improving the survival rate. PMID- 9356879 TI - Can optimal acid suppression prevent rebleeding in peptic ulcer patients with a non-bleeding visible vessel: a preliminary report of a randomized comparative study. AB - BACKGROUND/AIMS: The hypothesis that profound acid suppression might prevent clot lysis and thus benefit patients with a non-bleeding visible vessel has not been confirmed. Omeprazole can suppress gastric acid remarkably and may be beneficial for patients with peptic ulcer bleeding. METHODOLOGY: Fifty-two patients with a non-bleeding visible vessel at the ulcer base were enrolled and randomized into four groups (N = 13 in each group). In the cimetidine group, the patients received cimetidine 300 mg i.v. bolus followed by 300 mg i.v. every 6 hr during hospitalization. In the heater probe thermocoagulation + cimetidine group, the patients received heater probe thermocoagulation and cimetidine 300 mg i.v. bolus followed by 300 mg i.v. every 6 h during hospitalization. In the omeprazole q.d. group, the patients received omeprazole 40 mg i.v. bolus followed by 40 mg i.v. daily for two days. In the omeprazole q 12 h group, the patients received omeprazole 40 mg i.v. bolus followed by 40 mg i.v. every 12 h for two days. A 24 hr intragastric pH was recorded for every case. RESULTS: The mean 24 hr intragastric pH were higher in the omeprazole q.d. (mean 5.8) and the omeprazole q 12 h groups (mean 6.4) than in the cimetidine (mean 4.3) and the heater probe thermocoagulation + cimetidine groups (mean 4.9) (p < 0.05). Rebleeding occurred in 5, 2, 2 and 2 patients in the cimetidine, heater probe thermocoagulation + cimetidine, omeprazole q.d., and omeprazole q 12 h groups, respectively (p > 0.05). Volume of blood transfusion and number of days in hospital were not statistically different among the four groups. CONCLUSIONS: Omeprazole can remarkably suppress gastric acid when it is compared to that of the H2 receptor blocker. Patients with a non-bleeding visible vessel using omeprazole do not exhibit a decrease in the rebleeding rate as compared with those patients using cimetidine. PMID- 9356880 TI - Effect of hyperthyroidism on antral myoelectrical activity, gastric emptying and dyspepsia in man. AB - BACKGROUND/AIMS: The objective of the present study was to investigate the effect of hyperthyroidism on antral myoelectrical activity, gastric emptying and dyspepsia in man. METHODOLOGY: Twenty-three patients with manifest hyperthyroidism and dyspepsia confirmed by a standardized protocol were studied by electrogastrography (EGG). The following EGG parameters were determined: dominant frequency (DF cycles per minute (cpm), DF (%) in the normal range (2-4 cpm)), bradygastria (< 2 cpm), tachygastria (4-10 cpm), dominant frequency instability coefficient (DFIC), and postprandial to fasting power ratio. Data were correlated to results obtained in 18 age- and gender-matched controls. In 10 patients, a control measurement was performed after antithyroid therapy. In addition, in 15 consecutive patients, EGG data were compared to gastric emptying of radionuclides recorded simultaneously (gamma camera). RESULTS: Hyperthyroid patients revealed a significant increase in preprandial DF, and in pre- and post prandial tachygastrias compared to controls (3.3 cpm vs 3.1 cpm; 8.8% vs 3.5%; 12.3% vs 3.5%; p < 0.05). After antithyroid therapy, postprandial tachygastrias were reduced significantly. About 50% of the patients exhibited delayed gastric emptying compared to previously evaluated normal values (t 60 nuclide retention: > 68%). However, these patients did not differ in tachygastria and the other EGG parameters from those with normal gastric emptying (p > 0.05). Both EGG and radioscintigraphy did not correlate significantly with dyspepsia. CONCLUSIONS: Dyspeptic patients with hyperthyroidism frequently display tachygastria and delayed gastric emptying. However, tachygastria has no important effect on gastric motor activity in hyperthyroidism. PMID- 9356881 TI - Risk factors of esophagojejunal anastomotic leakage after total gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: The purpose of this study was to investigate the incidence of esophagojejunal anastomotic leakage (EJAL) after total gastrectomy. METHODOLOGY: Four hundred and four consecutive gastrectomy cases were reviewed to determine the incidence of esophagojejunal anastomotic leakage. RESULTS: EJAL developed in 33 patients (8.2%). The rate of leakage was found to be significantly related to the preoperative lymphocyte count and serum albumin level. Cases of para-aortic lymph node dissection (D4) had a significantly higher rate (16.1%) of EJAL than in conventional lymph node dissection (D2,3: 5.3%). The left upper abdominal evisceration group demonstrated a significantly higher EJAL rate (20.0%) than the cases without combined resection (4.8%). CONCLUSION: Aggressive surgery for advanced gastric cancer increases the risk of esophagojejunal anastomotic leakage. When aggressive surgery is necessary for curative purposes, meticulous preoperative, intraoperative and postoperative care are indispensable. PMID- 9356882 TI - Primary non-Hodgkin lymphoma of the stomach--role of surgery. AB - Thirty-one patients with Non-Hodgkin lymphoma of the stomach were retrospectively investigated. The prognostic factors regarding survival after treatment were determined in an univariate analysis. Twenty-seven of the patients revealed a stage I or II disease (according to the Ann Arbor classification) at the time of operation. The analysis showed no significance for grading, staging, lymph node-, serosa- or splenic involvement of our patients. Significant prognostic factors for survival of gastric Non-Hodgkin lymphomas were age (< 60 years) and resection margins R0 or R1/R2 (p < 0.05). The post-treatment evaluations for prognostic differentation between radical resections, patients with complete response after palliative resection including additional chemotherapy and patients without therapy response showed a significant difference in survival (p < 0.01). PMID- 9356883 TI - Plasma interleukin-6 is not a mediator of changes in lipoprotein lipase activity in cancer patients. AB - BACKGROUND/AIMS: Cancer cachexia is characterized by a variety of metabolic disorders. Alterations in fat metabolism have been reported to be associated with suppression of tissue lipoprotein lipase (LPL) activity in tumor-bearing animals. Interleukin-6 (IL- 6) has been documented to reduce tissue LPL activity and may play a role in inducing cancer cachexia. This study was conducted to clarify the changes in LPL activity and the role of IL-6 in patients with either gastrointestinal cancer or breast cancer. METHODOLOGY: Twelve patients with colorectal cancer, 7 patients with gastric cancer, 7 patients with breast cancer and 5 normal volunteers were studied. Serum concentrations of triglycerides (TG), non-esterified fatty acids (NEFA) and IL-6 were measured. LPL activity was measured in plasma post-heparin administration. The relationships of LPL activity to tumor progression, body weight loss and serum IL-6 levels were examined. The effect of tumor resection on LPL activity was also studied. RESULTS: LPL activity was suppressed with tumor progression in patients with either gastrointestinal cancer or breast cancer. Suppression of LPL activity and the degree of weight loss were negatively correlated in patients with either gastric or colorectal cancer (r = -0.5826, p = 0.011) but not in patients with breast cancer. The decrease in LPL activity was not always reversed after resection of the tumor. Circulating IL-6 did not correlate with either plasma LPL activity or tumor progression. CONCLUSIONS: Reduced LPL activity in patients with advanced gastrointestinal or breast cancer may reflect changes in nutritional status. Serum IL-6 is less likely to be a mediator of these alterations. PMID- 9356884 TI - The long-term course of severe anorexia nervosa in adolescents: survival analysis of recovery, relapse, and outcome predictors over 10-15 years in a prospective study. AB - OBJECTIVE: To assess the long-term course of recovery and relapse and predictors of outcome in anorexia nervosa. METHOD: A naturalistic, longitudinal prospective design was used to assess recovery and relapse in patients ascertained through a university-based specialty treatment program. Patients were assessed semiannually for 5 years and annually thereafter over 10-15 years from the time of their index admission. Recovery was defined in terms of varying levels of symptom remission maintained for no fewer than 8 consecutive weeks. RESULTS: Nearly 30% of patients had relapses following hospital discharge, prior to clinical recovery. However, most patients were weight recovered and menstruating regularly by the end of follow-up, with nearly 76% of the cohort meeting criteria for full recovery. Relapse after recovery was relatively uncommon. Of note, time to recovery was protracted, ranging from 57-79 months depending on definition of recovery. Among restrictors at intake, nearly 30% developed binge eating, occurring within 5 years of intake. A variety of predictors of chronic outcome and binge eating were identified. There were no deaths in the cohort. CONCLUSION: The course of anorexia nervosa is protracted. Predictors of outcome are surprisingly few, but those identified are in keeping with previous accounts. The intensive treatment received by these patients may account for the lower levels of morbidity and mortality when considered in relation to other reports in the follow-up literature. PMID- 9356885 TI - Six-year course of bulimia nervosa. AB - OBJECTIVE: Because little is known about the longer-term course of bulimia nervosa, the 2- and 6-year course and outcome of 196 consecutively treated females with bulimia nervosa-purging type (BN-P) was assessed. METHOD: One hundred ninety-six females with BN-P were assessed longitudinally at four points of time: at the beginning of therapy, at the end of therapy, at 2-year follow-up, and at 6-year follow-up. Self-rating scales as well as expert ratings from interview data were used. Eating disorder specific and general psychopathology was assessed. RESULTS: The general pattern of results over time showed substantial improvement during therapy, a slight (in most cases nonsignificant) decline during the first 2 years after treatment, and further improvement and stabilization from 3 to 6 years posttreatment. At 6-year follow-up, 20.9% had BN P, 0.5% BN-nonpurging type (BN-NP), 1.1% had shifted from BN to binge-eating disorder, 3.7% had anorexia nervosa, 1.6% were classified as eating disorder not otherwise specified (ED-NOS), and 2 patients had died; obesity with a body mass index (BMI) of > 30 was seen in 6.0%; the majority (71.1%) showed no major DSM-IV eating disorder. CONCLUSIONS: Based on a composite global outcome score at 6 years follow-up, 59.9% achieved a good outcome, 29.4% an intermediate outcome, 9.6% a poor outcome, and 2 (1.1%) persons were deceased. Course and outcome were generally more favorable than in anorexia nervosa. PMID- 9356886 TI - Beyond body image: the integration of feminist and transcultural theories in the understanding of self starvation. AB - OBJECTIVE: The present study represents an intersection between cross-cultural theorizing and teminist scholarship. It is an attempt to provoke as well as augment prevailing biomedical models that esteem fear of fatness as the primary motivation for voluntary starvation in anorexic women. METHOD: Recent studies of eating disturbance in both Eastern and Western societies are invoked to demonstrate the ways in which women straddling two worlds, be it generational, work-family, cultural or traditional and modern, may employ food denial as an instrumental means of negotiating the transition, disconnection, and oppression that they uniformly endure. RESULTS: A feminist/transcultural interpretation of the literature suggests that by construing anorexia nervosa as a body image disorder or Western culture-bound syndrome, extant models miss the broader contexts and varied meanings of food refusal. DISCUSSION: The implications of cross-disciplinary perspectives for theory building and treatment are discussed, acknowledging not only the gendered nature of eating disorders but their embodiment of power differentials as well. PMID- 9356887 TI - A comparison of ever-married and never-married women with anorexia nervosa or bulimia nervosa. AB - OBJECTIVE: The results of the scant research on anorexia nervosa and marriage suggest that married anorexics may exhibit more severely disordered eating. However, past research has not controlled for the greater age of married versus unmarried anorexics, and very little research has been conducted on marriage and women with bulimia nervosa. We investigated differences in disordered eating and clinical traits between ever-married and never-married women with anorexia nervosa or bulimia nervosa and statistically controlled for age. METHOD: Adult women ages 20-45, who were assessed in an outpatient eating disorders clinic and diagnosed with anorexia nervosa (n = 91) or bulimia nervosa (n = 223), completed several measures of disordered eating and related traits at the point of initial evaluation. RESULTS: In simple comparisons, ever-married women differed from their never-married peers with regard to several indices of symptom history and severity. However, after controlling for age, ever-married women differed only with regard to an earlier onset of menarche and, for women with bulimia nervosa, an earlier onset of sexual intercourse. DISCUSSION: Results are discussed with regard to possible explanation and directions for future research. PMID- 9356888 TI - Food cravings in women with a history of anorexia nervosa. AB - OBJECTIVE: The objective of the present study was to determine the prevalence and characteristics of food cravings in women with a history of anorexia nervosa. METHOD: One hundred one control women selected at random and 64 women with a diagnosis of anorexia nervosa 10 to 14 years earlier (cases) completed the Diagnostic Interview for Genetic Studies, a food craving questionnaire, the Temperament and Character Inventory and the Three-Factor Eating Questionnaire. RESULTS: A similar proportion of cases and controls reported food cravings. A greater proportion of cases reported strong cravings with two or more features of intensity (p = .02). Cravings in the cases were more likely to be characterized by difficulty resisting the craved food (p = .0008), anxiety when the craved food was unavailable (p = .002), and a high frequency of occurrence (p = .001). The cases who craved were significantly more likely to have had lifetime BN (p = .02). CONCLUSION: A similar prevalence of food craving in cases as in controls suggests that successful control of food intake and/or denial of hunger overrides dietary restriction as a precondition for craving in anorexia nervosa. A dysfunction in the serotonergic system, the provision of intermittent reinforcement by binge episodes, and/or frustration due to unsuccessful attempts at dietary restraint may mediate the association of cravings with the presence of lifetime bulimia. PMID- 9356889 TI - Persistence of binge-eating patterns after a history of restriction with intermittent bouts of refeeding on palatable food in rats: implications for bulimia nervosa. AB - OBJECTIVE: To test the hypothesis that experience with food restriction produces persistent binge eating. The Minnesota semistarvation experiment and studies of prisoners-of-war show that chronic food restriction produces dramatic changes in eating behavior (including binge eating) that endure decades after restriction has ceased. Bulimia nervosa patients who restrict also binge. Restriction may be a risk factor in the etiology of binge eating and bulimia. METHOD: Animals were subjected to four different patterns of 12-week restriction-refeeding cycles. The rats were either food restricted (dieting) or not restricted and refed regular or palatable food (binging). RESULTS: Thirty days after normalization (full feeding, no restriction cycling), rats with a history of cycles of restriction and hyperphagia continued to exhibit persistent binge eating. This effect was shown particularly with palatable food, in stated conditions, and in response to acute 24-hr deprivation. DISCUSSION: Results from this animal model implicate restriction and overeating on palatable food as biological determinants of binge eating behaviors, including bulimia nervosa. PMID- 9356890 TI - Normal and neurotic perfectionism in eating disorders: an interactive model. AB - OBJECTIVE: Previous nonclinical research found that both normal (or adaptive) and neurotic (or maladaptive) perfectionism were related, in the positive direction, to attitudes and behaviors associated with eating disorders. However, based on a related body of research, it was hypothesized that these two aspects of perfectionism would relate to body esteem in an interactive rather than an additive fashion. METHOD: Anorexic and bulimic patients (n = 123) were assessed on multidimensional aspects of perfectionism, neuroticism, and body esteem. RESULTS: Predictions were confirmed. Normal perfectionism was positively associated with body esteem, but only when levels of neurotic perfectionism were low. Conversely, body-image disparagement was most pronounced when normal and neurotic perfectionism were both elevated. DISCUSSION: These findings demonstrate that the interpretation of simple relationships among personality variables--at least in the area of body image and eating disorder research--may provide a misleading picture. PMID- 9356891 TI - Stress, coping, and disturbed eating attitudes in teenage girls. AB - OBJECTIVE: This study explored the relationship between stressors and disturbed eating attitudes among adolescent females, assessing the moderating role of coping and the mediating influence of poor self-esteem. METHOD: Two hundred eighty-six teenage girls were recruited from local schools, and completed standardized measures of stressors, coping, self-esteem, perfectionism, and disturbed eating attitudes. Regression analyses were used to test for moderating and mediating effects. RESULTS: Stressors and emotion-focused coping were found to be associated with low self-esteem, which in turn was strongly associated with disturbed eating attitudes. Stressors were also directly related to disturbed eating attitudes. DISCUSSION: The findings provide partial support for existing models of the etiology and maintenance of eating psychopathology, but have wider implications for our understanding of the eating disorders and their treatment. PMID- 9356892 TI - Self-reported dieting: how should we ask? What does it mean? Associations between dieting and reported energy intake. AB - OBJECTIVES: To determine whether self-reports of dieting to control weight are associated with reported energy intake, how this association varies with the phrasing of questions on dieting behaviors, and whether this association differs by educational level and weight status among adult male and female respondents. METHODS: The study population included 996 women and 227 men, aged 20-45, who volunteered to participate in a weight gain prevention trial. Participants completed surveys at baseline regarding their dieting behaviors and nutritional intake. RESULTS: The association between self-reported dieting and energy intake varied according to the phrasing of the questions assessing dieting behaviors. Multi-item scales and nonambiguous single-item questions (e.g., "current dieting") were more strongly associated with reported energy intake than more general single-item questions (e.g., "doing anything to lose weight"). Overweight dieters reported lower energy intake than overweight nondieters. Among nonoverweight persons, associations between dieting and energy intake were not significant. The association between dieting and energy intake did not differ by educational level among women. Among men, dieting predicted lower energy intake in those with low educational levels, although the number of men with low educational levels who were dieting was small. Reported dieting was not associated with energy intake among men with higher educational levels. DISCUSSION: Associations between self-reports of dieting and reported energy intake vary according to the phrasing of specific questions about dieting, gender, education, and weight status. These factors should be taken into account in the design of instruments for measuring these behaviors and in the interpretation of results, especially across studies using different methodologies. PMID- 9356893 TI - Patients with chronic fatigue syndrome and accurate feeling-of-knowing judgments. AB - Many Chronic Fatigue Syndrome (CFS) patients complain of memory impairments which have been difficult to document empirically. Subjective complaints of memory impairment may be due to a deficit in metamemory judgment. CFS patients and matched controls were tested with a computerized Trivia Information Quiz that required them to rate their confidence about correctly recognizing an answer in a multiple choice format that they had been unable to remember in a fact-recall format. Even though CFS patients reported significantly greater amounts of fatigue, cognitive, and physical symptoms, the accuracy of their confidence levels and recognition responses were similar to controls. This finding suggests that a metamemory deficit is not the cause of the memory problems reported by CFS patients. PMID- 9356894 TI - Estimating premorbid WAIS-R intelligence in the elderly: an extension and cross validation of new regression equations. AB - Recently developed equations that combine WAIS-R subtests and demographic information to estimate premorbid intelligence were evaluated in persons 75 years and older. The equations underestimated the obtained IQs of elderly persons. Therefore, new equations were developed using the old age WAIS-R standardization sample. The new equations cross-validated well and demonstrated adequate ability to detect possible intellectual deterioration in a brain damaged sample. The demographic formulas of Barona, Reynolds, and Chastain worked as well as the new equations in suggesting possible deterioration in a sample of elderly persons with chronic and diffuse cognitive impairment. PMID- 9356895 TI - Which tests do neuropsychologists use? AB - A nationwide survey of accredited neuropsychologists in Australia was conducted to examine test use. Clinicians were asked to list the tests they give most often. Results are expressed as endorsement frequencies for tests. Comparisons with international surveys of test use are provided. Suggestions for clinicians and others concerned with test use are included to demonstrate how survey results can be used to improve neuropsychological services. PMID- 9356896 TI - Rey Auditory-Verbal Learning Test: structure of a modified German version. AB - The Auditory Verbal Learning Test (AVLT) is widely used in scientific research as well as in clinical practice. But there exists little research on the structure of the AVLT. We investigated the structure of a German version of the AVLT and VLMT, in 232 patients of a psychiatric clinic and in 872 patients of an epileptologic clinic. First we stated a theoretical LISREL model relating the observed variables of the VLMT to short-term memory (STM) and long-term memory (LTM) as latent variables. Then we estimated the postulated LISREL model in the two samples. The proposed model showed excellent fit in both samples, and there were no significant deviations between the estimated and the observed covariance matrices. Thus, STM and LTM suffice to explain the structure of the VLMT, and the proposed structural equations model can be used to estimate STM and LTM capacity from VLMT data. PMID- 9356897 TI - Longitudinal and psychometric profiles of two cognitive status tests in very old adults. AB - Forty-four nondemented adults, over the age of 75 years, were tested at six-month intervals spanning two years. Study goals were to examine the validity of the Mini-Mental State Examination (MMSE) and the Wechsler Adult Intelligence Scale Revised (WAIS-R), to assess the long-term reliability of these instruments, and to examine the longitudinal profile of this sample. Results showed that the MMSE was moderately correlated with the WAIS-R. The MMSE had low internal consistency, although the total score behaved reliably across the five occasions. The WAIS-R scales showed a high degree of internal consistency and test-retest reliability. WAIS-R performance remained stable across occasions; however, decline was evident in the MMSE subtests of Attention and Language. PMID- 9356898 TI - Norms for an abbreviated Raven's Coloured Progressive Matrices in an older sample. AB - Percentile age norms for ages 55 to 85 using overlapping intervals at specified age midpoints are presented for the sum scores of sections A and B of Raven's Coloured Progressive Matrices (RCPM). The representative age and gender stratified sample (N = 2,815) used is derived from the Longitudinal Aging Study Amsterdam (the Netherlands). As RCPM scores appear to be strongly associated with education, percentile norms for three educational levels are presented: low (0-9 years), middle (10-15 years) and high (16 years and more). PMID- 9356899 TI - Neurotoxicity of chronic low-dose exposure to organic solvents: a skeptical review. AB - The health effects of long-term, low-level exposure to organic solvents have been studied for many years. While the volume of literature is great, definitive conclusions regarding chronic neurobehavioral effects of environmental exposure are premature. Methodological shortcomings in research preclude confidence in studies allegedly supporting a causal link between chronic low-dose solvent exposure and lasting neurobehavioral deficits. In this article, the shortcomings reviewed include selection bias in recruitment of research subjects, overreliance on subjective recall in determining levels and duration of exposure, between study variability in kinds of solvents examined, variability in tests selected to assess neurobehavioral functioning, and diversity in reported findings. The implications of these for characterizing the state of organic solvent research are discussed. PMID- 9356900 TI - Comparison of NART and Barona demographic equation premorbid IQ estimates in Alzheimer's disease. AB - Two methods of estimating premorbid WAIS-R intelligence were compared in matched samples of normal and AD persons. The NART and Barona 1984 demographic equations accurately predicted the IQs of the normal group and overestimated the IQs of the AD subjects. When the AD group was divided into mild and moderately impaired subgroups, the more severely demented subjects displayed lower WAIS-R IQs and NART estimated IQs, revealing that NART performance is sensitive to dementia severity. However, the NART estimated IQs for the mild and moderately impaired AD subgroups were larger than the WAIS-R IQs, suggesting that while the NART is sensitive to dementia severity, it may still provide relevant clinical information. PMID- 9356901 TI - Construct validity of the computerized version of the Category Test. AB - The purpose of the current study was to investigate the construct validity of the computer version of the Category Test (CT) in relation to the standard version. As part of a comprehensive neuropsychological assessment, outpatient rehabilitation clients completed either the standard CT (n = 49) or the computerized version (n = 56). Two principal component factor analyses were performed and the factor structures compared to a solution reported previously in the literature. Inspection of the factor loadings revealed a similarity between the computerized CT and standard CT groups as well as with the previously reported analysis. The results of this study suggest that regardless of the mode of administration the CT loads on a "spatial abilities", factor and provides support for the construct validity of the computerized CT as part of a comprehensive neuropsychological testing battery. PMID- 9356902 TI - Construct validity of the MMPI-168(L) with mentally retarded adults and adolescents. AB - Fifty-three persons residing in an institution and diagnosed with mild or moderate mental retardation were assessed with a modification of the MMPI-168. Forty-one of the residents also had psychiatric diagnoses. Construct validity of the MMPI-168(L) was examined by correlating T scores obtained on the scales of this instrument with the results of an 8-item "Behavioral Survey." The survey required unit clinicians to rate the severity of behavioral disturbance in eight categories. Six of the items on the "Behavioral Survey" were found to correlate with one or more MMPI scales. Stepwise multiple regression procedures revealed additional collective relations between elevations on the MMPI and ratings of behavioral disturbance. PMID- 9356903 TI - The Neurobehavioral Cognitive Status Examination: accuracy of the "screen-metric" approach in a clinical sample. AB - This study assessed the accuracy of the "screen" versus "metric" portions of eight subtests of the Neurobehavioral Cognitive Status Examination (NCSE). As part of a routine hospital assessment, 95 male patients were administered both portions of the instrument regardless of outcome on the screen. Results indicate that the screen items of some of the NCSE subtests produced a relatively high false negative rate, where the screen was passed, but the metric was failed. It is recommended that all items of the subtests be administered to more fully assess each domain and, therefore, reduce the probability of overlooking significant deficits. PMID- 9356904 TI - EEG correlates of a paper-and-pencil test measuring hemisphericity. AB - The present study examined the validity of the Preference Test (PT), a questionnaire that intends to measure the extent to which subjects rely on right hemisphere and left-hemisphere cognitions. Normal subjects (N = 20) completed the PT and were then assigned to a group with a relatively strong preference for left hemisphere cognitions or a group with a relatively strong preference for right hemisphere cognitions. Next, background EEG was recorded. Subjects with a preference for right-hemisphere cognitions were found to display relatively more alpha power over the left than over the right frontal areas, compared to subjects with a preference for left-hemisphere cognitions. This finding provides partial support for the validity of the PT. PMID- 9356905 TI - MMPI-2 base rates for 492 personal injury plaintiffs: implications and challenges for forensic assessment. AB - This study reports base rates of MMPI-2 clinical scales. PTSD scales, and validity scales for 492 personal injury plaintiffs, 230 men and 262 women. Scales studied included L, F, K, F minus K, Ds-r, Fake Bad, Ego Strength, Back F. Total Obvious minus Subtle, VRIN, and TRIN. Forensic high points resembled outpatient profiles but not the MMPI-2 psychiatric sample and shared only code type 13/31 with the normative sample. The most common two-point code type for men was 13/31, followed by 12/21 and 23/32, and for women was 13/31, followed by 23/32 and 12/21. Fifty percent of the forensic sample were code type 13/31, 12/21, or 23/32. Validity measures suggested possible malingering on approximately 20 to 30% of the profiles but the majority of profiles were valid. Validity problems discussed include attorney coaching and the congruence of plaintiff personality characteristics with the demand characteristics of litigation. Examples of attorney coaching are provided. The modal plaintiff appears to be an unhappy somatizer involved in a social context which encourages rationalization, projection of blame, and complaining. PMID- 9356906 TI - Rey II: redesigning the Rey screening test of malingering. AB - This study has redesigned the Rey 15-item Visual Memory Test (1964) by introducing more complex figures and by increasing internal logic and pattern redundancy. Standardized administrative procedures and rules for a simple qualitative scoring system were established. Performance on the original Rey continued to be significantly contaminated by ability components and illness while performance on the Rey II qualitative scoring system was not significantly related to intelligence, age, mental status or memory. The Rey II demonstrated improved face validity. Linear Discriminant Function Analysis indicated that the qualitative scoring system had a higher classification accuracy than the quantitative system on both instruments; the Rey II qualitative scoring system accurately detected 31% more college malingerers than the Rey quantitative and 21% more clinical malingerers than the Rey II quantitative. A malingering cut-off of two qualitative errors gave the Rey II a 79% higher sensitivity in the college malingerers and 29% higher specificity in the clinical population than the standard quantitative Rey cut-off of nine items. PMID- 9356907 TI - Acquired immunodeficiency syndrome dementia complex. AB - Acquired immunodeficiency syndrome (AIDS) has become an epidemic in the United States. AIDS dementia complex (ADC) is a neurological dysfunction which has been indicated in 25-90% of AIDS patients, 30-40% of HIV-infected patients, and may be the only presenting manifestation of AIDS. Researchers have investigated many aspects of ADC including clinical features, etiology, epidemiology and prevalence, diagnosis (psychological parameters and laboratory investigations such as CSF, EEG, CT, MRI, PET, and ERP), assessment, neurological features (including neuropsychiatric and neuropsychological measures, and neuropathology), prognosis, and treatment. The research is controversial, complex, and contradictory. A discussion of the many areas of ADC and many hypotheses will be included. PMID- 9356908 TI - The cost of growth: our professional crossroads. Educational Enhancement Project- an overview. PMID- 9356910 TI - Long-term efficacy of total SILASTIC implants: a subjective analysis. AB - A retrospective analysis of the long-term efficacy of total SILASTIC implant arthroplasty performed before 1986 is presented. A total of 50 patients responded to subjective questionnaires regarding pain, function, complications, and overall patient satisfaction. The average age of the patients at the time of surgery was 55.1 years with an average follow-up of 13.4 years (range 10.7 to 16.9 years). Ninety-seven percent of patients reported relief from pain, and the overall success rating was 90.7%. Results were calculated based on a modification of the American Orthopaedic Foot and Ankle Society clinical rating system; the mean rating was 87.3. Attention must be directed at realigning the joint via appropriate osteotomies and soft tissue balancing procedures for increased success. Although radiographic deterioration of the implant was demonstrated in all implants, this deterioration did not correlate with patient satisfaction and should not be the sole criterion for implant removal. We conclude that total implant arthroplasty is a proven procedure for long-term relief of pain in selected patients with degenerative joint disease of the first metatarsophalangeal joint. PMID- 9356909 TI - The role of cheilectomy in the treatment of hallux rigidus. AB - The aim of this study was to evaluate and compare the outcome of cheilectomy for patients with grades 1, 2, and 3 hallux rigidus according to Regnauld's classification. We evaluated 39 patients at a mean of 3.8 years after surgery for pre- versus postoperative changes in pain, activity level, difficulties with footwear, tiptoe walking, and metatarsophalangeal joint range of motion. We used Wilcoxon paired rank tests to compare outcomes for each grade of hallux rigidus. There was significant improvement (p < 0.05) in pain, activity level, tiptoe walking, and range of motion for all three grades of hallux rigidus. Footwear selection was significantly improved in Grades 1 and 2 patients but not in patients with Grade 3 hallux rigidus. PMID- 9356911 TI - The nonfixated Austin bunionectomy: a retrospective study of one-hundred procedures. AB - A retrospective evaluation of 64 randomly selected patients with 100 nonfixated Austin bunionectomy procedures was performed. A radiographic and a clinical evaluation were performed, including an analysis of preoperative and postoperative angles as well as postoperative complications. This study demonstrates a similar complication rate for nonfixated Austin bunionectomies as compared with previous studies with internal fixation. Removal of the fibular sesamoid was performed in 90% of the cases and did not increase the incidence of hallux varus. The nonfixated Austin bunionectomy is an acceptable alternative to the correction of hallux valgus. If internal fixation is utilized, the most cost effective device should be used. PMID- 9356912 TI - Our fixation with fixation: are screws clinically superior to external wires in distal first metatarsal osteotomies? AB - The purpose of this study was to evaluate the immediate postoperative morbidity, the structural correction attained, and the long-term range of motion following fixation with a single external Kirschner wire and an internal cortical screw. We abstracted records for 69 patients undergoing, distal unicorrectional chevron osteotomies. Thirty-three patients received percutaneous 0.062-inch K-wire fixation and 36 patients received single 2.7-mm. cortical screw fixation. Among these age- and sex-matched subjects, there was not a significant difference between any of the correctional or morbid outcomes measured in this study on the basis of type of fixation employed. Patients with rigid internal screw fixation did not return to shoe gear sooner, develop fewer postoperative infections, or have increased long-term range of motion than the group receiving external fixation with a single K-wire. Surgical time was significantly longer for those patients undergoing rigid internal fixation with a screw (42.5 +/- 9.5 vs. 35.1 +/- 6.6 minutes, p < 0.001). We conclude that there is no significant difference in postoperative infection, dehiscence, long-term structural correction attained, or range of motion achieved between rigid internal screws and external K-wires used to fixate distal metatarsal osteotomies. PMID- 9356913 TI - Modified Jones procedure for post-polio claw hallux deformity. AB - A prospective study was carried out on 12 patients with postpoliomyelitis clawing of the hallux treated by a modified Jones procedure using Kirschner wire for the interphalangeal joint fusion. The purpose of this study was to evaluate the outcome of a modified Jones procedure in the treatment of postpolio claw hallux deformity. All the patients had symptoms related to the claw hallux deformity; foot biomechanics and gait were affected. Patients were assessed both pre- and postoperatively using Axer's criteria. Mean follow-up was 32 months. Ten patients had very good results and two patients had fair results. The transferred extensor hallucis longus became loose in two patients who had a coexistent tight Achilles tendon. Recurrence of medial cavus deformity occurred 3 months after the operation in both of these patients. This was treated by shortening the transferred extensor hallucis longus after elongating the tight Achilles tendon. No pseudoarthrosis of the interphalangeal joint was identified. When the Jones procedure is performed, the motor power of the extensor hallucis longus should be Medical Research Council Grade V before transfer. The Achilles tendon should be evaluated for equinus preoperatively and lengthened when equinus deformity is present in order to avoid residual foot deformities. PMID- 9356914 TI - Neuropathic ulcerations plantar to the lateral column in patients with Charcot foot deformity: a flexible approach to limb salvage. AB - Neuroarthropathy of the midfoot may lead to a structural deformity that predisposes the diabetic patient to skin breakdown and ulceration. In some cases, conservative management is not adequate, making surgical intervention necessary. The authors performed a retrospective study to look at those patients who required surgical intervention for a specific pattern of neuroarthropathy. Over a 2 1/2-year period, 32 feet (31 patients) underwent surgical procedures for treatment of nonhealing neuropathic ulcerations beneath the lateral column of Charcot feet. All feet underwent exostectomy with 17 undergoing excision of the ulcer with primary closure, 8 closure via rotational fasciocutaneous flap with transpositional intrinsic muscle flap, and 6 through an incision placed adjacent to the ulcer. One patient whose ulcer was healed at the time of surgery had the incision placed directly over the prominence. Overall, 29 of 32 feet maintained functional limb salvage. This included eight patients who required revisional surgery, either by resection of more bone or creation of a local flap for coverage. Life-table analysis resulted in an 89% overall success rate. The results show that a flexible approach to skin and soft tissue coverage is necessary to heal these patients, provided attention is directed to the underlying bony prominence. PMID- 9356915 TI - Sonographic evaluation of interdigital neuromas. AB - The purpose of this study is to evaluate the effectiveness of sonography to accurately identify interdigital neuromas. Twenty patients, all complaining of interdigital neuroma type pain, underwent sonographic evaluation of the symptomatic intermetatarsal space and adjacent interspaces. Each patient subsequently had surgical exploration of the symptomatic site. Fourteen patients had sonographic evidence of neuromas which were confirmed surgically; one patient had sonographic evidence of a neuroma that was not confirmed surgically. The remaining five patients did not have sonographic or surgical evidence of neuroma in spite of clinical signs and symptoms. No adjacent neuromas were appreciated. The correlation of sonography and clinical measurements was determined using Pearson's product moment correlation coefficient (r = 0.804, p < 0.001). Measurements performed with ultrasound were highly correlated with surgical findings. The mean size (standard deviation) of neuromas identified with ultrasound and during surgical inspection was 5.35 mm. (1.36) and 5.43 mm. (1.04). Ultrasound demonstrated a sensitivity of 100% and a specificity of 83.3%. Ultrasound was able to accurately predict the presence, size, and location of Morton's neuromas. PMID- 9356916 TI - Human articular cartilage biomechanics of the second metatarsal intermediate cuneiform joint. AB - Intrinsic material properties and histomorphometry of freshly frozen, human cadaveric cartilage from the second metatarsal intermediate cuneiform (SMIC) articulation were obtained to provide biomechanical mapping of the surfaces. The biphasic creep indentation methodology and an automated creep indentation apparatus were used to measure aggregate modulus, Poisson's ratio, permeability, shear modulus, and thickness. Biomechanical experiments were performed on four sites of the SMIC joint in 14 specimens (seven pairs): two sites in the second metatarsal base and two sites in the intermediate cuneiform head. Results of the study indicate that no significant variations exist in the biomechanical comparisons between specific articulations, gross articulations, and left and right joints. For example, cartilage from the second metatarsal base and intermediate cuneiform head had an aggregate modulus of 0.99 MPa and 1.05 MPa, respectively. The Poisson's ratio and permeability of all test sites grouped together were found to be 0.08 and 3.05 x 10(-15) m4/N.s, respectively. Cartilage thickness was measured at 0.61 mm. This biomechanical study suggests that similarities in cartilage properties may be beneficial in preventing the human SMIC articulation from developing early degenerative changes. Histological evaluation demonstrated that SMIC cartilage exhibits structural characteristics (such as the absence of chondrocyte columnar arrangement in the deep zone) which may be typical of cartilage that does not experience habitually high compressive stresses. This knowledge could aid surgeons in generating a deeper perspective of the relationship between clinical pathologies of articular cartilage and intrinsic biomechanical etiologies of degenerative joint diseases. PMID- 9356917 TI - Repair of an osteochondral tumor of the talus utilizing a fresh-frozen cadaveric graft. AB - A case involving a benign osteochondral tumor of the talar dome is presented. Graft considerations are reviewed and some of their specific characteristics are highlighted. Selection of a fresh-frozen cadaveric graft in this case was based on the goal of maintaining articular congruity in the ankle joint. The authors' experience demonstrated that normal joint function can be preserved with this type of graft. PMID- 9356918 TI - Podiatric surgical considerations in the Ehlers-Danlos patient. AB - Ehlers-Danlos syndrome is a relatively common, heritable disorder of connective tissue with multiple manifestations affecting the lower extremities. A review of the world literature reveals limited documentation of podiatric surgical procedures performed on this unique group of patients. Potential podiatric surgical complications in the Ehlers-Danlos syndrome patient population are derived from a thorough review of previously performed surgical procedures. PMID- 9356919 TI - Rupture of the medial collateral ligament of the first metatarsophalangeal joint in a professional soccer player. AB - Worldwide, more people play soccer than any other team sport. The Federation Internationale de Football Association (FIFA) registered more than 150 million players in 1984. Although foot injuries in soccer range from midfoot sprains to stress fractures to capsulitis of the first metatarsophalangeal joint, we could find no case reports of a rupture of the lateral collateral ligaments of the great toe in soccer players. This is a report of the diagnosis, treatment, and outcome of such an injury in a professional soccer player. PMID- 9356920 TI - The use of the offset blade system in foot surgery. PMID- 9356921 TI - Osteonecrosis of the tibial and fibular sesamoids in an aerobic dancer. PMID- 9356922 TI - Distribution of adrenocorticoid receptors in the rat CNS measured by competitive PCR and cytosolic binding. AB - Combined quantitative polymerase chain reaction (PCR) and cytosolic binding assay techniques are used to measure mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) mRNA, Kd, and Bmax in various rat central nervous system (CNS) regions, namely amygdala, hypothalamus, hippocampus, cortex, pituitary, and cervical, thoracic, and lumbar spinal cord. Two internal standards (i.s.) cDNA were cloned for quantitative PCR purposes. The i.s. templates differed from the respective wild-type (wt) templates for a single base-pair mutation introduced by PCR that generated a unique restriction site, thus allowing amplification products arising from coamplification of wt and i.s. to be distinguished. Results show that cerebellum, which displayed average Bmax values for both receptors, contained the highest level of MR and GR mRNA. Hippocampus also had a high level of MR mRNA. Low mRNA content was found in the hypothalamus for MR and GR as well as in the cortex for GR. High Bmax values for both MR and GR were found in the lumbar spinal cord, despite a modest mRNA content. The lowest Bmax values were found in the cortex for both receptors. It is, therefore, concluded that mRNA content and Bmax are not closely correlated in the rat CNS. These data suggest a differential regulation of various adrenocorticoid receptor isoforms. Moreover, this quantitative PCR method is very sensitive and can be used to assay small amounts of material in order to obtain absolute measurements of mRNA expression. PMID- 9356924 TI - Expression of mRNA for Akt, serine-threonine protein kinase, in the brain during development and its transient enhancement following axotomy of hypoglossal nerve. AB - By in situ hybridization histochemistry, expression of mRNAs for the two species of serine/ threonine protein kinase Akt, Akt1 and Akt2, were examined in the mouse brain during normal development and in the hypoglossal nucleus following axotomy. On the embryonic days, the gene expression for Akt1 and Akt2 was detected at high levels throughout the entire neuroaxis, then decreased gradually to adult levels during postnatal development. In the adult brain, the gene expression for Akt1 and Akt2 was weak in almost all neurons with no difference of expression levels. The expression level of Akt1 mRNA in the affected hypoglossal nucleus increased dramatically after 48 h to 7 d following axotomy of the hypoglossal nerve, whereas no change was seen in the level of Akt2 mRNA. The present findings suggest that Akt may contribute some important roles not only in neurogenesis, but also in regeneration of injured neuron. PMID- 9356923 TI - Acidic and basic fibroblast growth factor messenger RNA and protein show increased expression in adult compared to developing normal and dystrophic rat retina. AB - To further elucidate the possible roles of fibroblast growth factors (FGFs) in retinal pathophysiology, messenger RNA levels of acidic and basic FGF (aFGF and bFGF, respectively) were measured throughout embryonic and postnatal development until adulthood in normal and dystrophic (Royal College of Surgeons, RCS) rat retinas using sensitive reverse transcription-coupled polymerase chain reaction (PCR) techniques. In normal rats, both aFGF and bFGF transcript levels remained steadily low throughout embryogenesis and up until 7 d of postnatal age. By 13 d bFGF mRNA had increased 30-fold, and by adulthood (4 mo) levels were 150 times greater than in newborn retina. Dystrophic RCS retinas followed the same basic pattern, except that bFGF expression levels were increased relative to normal rats: By 4 d postnatal RCS retinas contained three times more bFGF mRNA than normal, by 7 d they contained six times more, and by 10 d they contained eight times more. In contrast, aFGF mRNA levels rose only threefold between embryonic and adult stages, and did not show any differences between normal and RCS rats. In parallel, staining of lightly fixed frozen sections of young (< 20 d) normal rat retina with antibodies to bFGF revealed only faint labeling of neural cells, whereas adult retinal sections were labeled strongly, especially within the photoreceptor layer. Twenty-day RCS rat retina showed detectable bFGF-like immunoreactivity. Hence, these data indicate that major aFGF and bFGF expression occurs only late in retinal maturation, suggesting these factors act principally as survival factors, especially for photoreceptors. In addition, the increased expression in a degenerative mutant strain may indicate the early onset of general cellular stress. PMID- 9356926 TI - Lack of interactions between amyloid precursor protein and hydrophilic domains of presenilin 1 and 2 using the yeast two hybrid system. AB - Mutations in the two related genes, presenilin 1 (PS1) and presenilin 2 (PS2), which are predicted multispanning membrane proteins, are responsible for the majority of early-onset familial Alzheimer's disease (FAD). To demonstrate direct interactions between presenilins (PS) and amyloid precursor protein (APP), the authors utilized a yeast two-hybrid system. Various hydrophilic domains derived from PS and those of APP were coexpressed in yeast and tested for the interaction. No detectable interactions were found in any PS/APP set examined. The authors' studies suggest that PS and APP do not interact through their hydrophilic domains in yeast, raising the possibility that interaction may occur indirectly or require proper conformation or subunit formation. PMID- 9356925 TI - A chimeric tyrosine/tryptophan hydroxylase. The tyrosine hydroxylase regulatory domain serves to stabilize enzyme activity. AB - The neurotransmitter biosynthetic enzymes, tyrosine hydroxylase (TH), and tryptophan hydroxylase (TPH) are each composed of an amino-terminal regulatory domain and a carboxyl-terminal catalytic domain. A chimeric hydroxylase was generated by coupling the regulatory domain of TH (TH-R) to the catalytic domain of TPH (TPH-C) and expressing the recombinant enzyme in bacteria. The chimeric junction was created at proline 165 in TH and proline 106 in TPH because this residue is within a conserved five amino-acid span (ValProTrpPhePro) that defines the beginning of the highly homologous catalytic domains of TH and TPH. Radioenzymatic activity assays demonstrated that the TH-R/TPH-C chimera hydroxylates tryptophan, but not tyrosine. Therefore, the regulatory domain does not confer substrate specificity. Although the TH-R/TPH-C enzyme did serve as a substrate for protein kinase (PKA), activation was not observed following phosphorylation. Phosphorylation studies in combination with kinetic data provided evidence that TH-R does not exert a dominant influence on TPH-C. Stability assays revealed that, whereas TH exhibited a t1/2 of 84 min at 37 degrees C, TPH was much less stable (t1/2 = 28.3 min). The stability profile of TH-R/TPH-C, however, was superimposable on that of TH. Removal of the regulatory domain (a deletion of 165 amino acids from the N-terminus) of TH rendered the catalytic domain highly unstable, as demonstrated by a t1/2 of 14 min. The authors conclude that the regulatory domain of TH functions as a stabilizer of enzyme activity. As a corollary, the well-characterized instability of TPH may be attributed to the inability of its regulatory domain to stabilize the catalytic domain. PMID- 9356928 TI - A nuclear matrix attachment region is highly homologous to a conserved domain of olfactory receptors. PMID- 9356927 TI - cDNA cloning and mRNA expression analysis of the human neuronatin. High level expression in human pituitary gland and pituitary adenomas. AB - The authors cloned the nearly complete cDNA of human neuronatin with the aid of an expressed sequence tag (EST) database, and analyzed its expression in various human tissues by Northern blot analysis. The nucleotide and deduced amino acid sequences of the human neuronatin showed a high similarity to those of rodents. The Northern blot analysis revealed that the human neuronatin message was expressed predominantly in the fetal brain in the brain-specific manner, but only faintly in the adult brain. Among the various adult human tissues examined, the anterior pituitary gland was shown to be the only place where the neuronatin mRNA was strongly expressed. Intense neuronatin expression was also observed in several human pituitary adenomas, including ACTH-producing, GH-producing, and nonfunctioning adenomas, but hardly detected in other brain tumors. PMID- 9356929 TI - Influence of rotator cuff tearing on glenohumeral stability. AB - This study hypothesizes that full-thickness tearing of the rotator cuff can lead to joint instability and that the degree of instability depends on the size and location of the tear. Twelve cadaveric shoulder specimens were divided into two groups: group 1 had a circular tear centered at the critical area, and group 2 had a circular tear centered at the rotator interval. Each group was tested at 2.5 cm and 5 cm tear sizes. Unloaded, and with the arm in 90 degrees flexion and full internal rotation, the humeral head shifted posteriorly. With loading, a large and more anteriorly located defect had the most influence on stability. The tear size had the greatest effect on stability in the inferior direction for group 1 and on the anterior direction for group 2. The tear location had the most significant effect on stability in the inferior and anterior directions for the smaller tear and on the anterior direction for a larger tear. PMID- 9356930 TI - The influence of intramedullary fixation on figure-of-eight wiring for surgical neck fractures of the proximal humerus: a biomechanical comparison. AB - The resistance to torsional load was measured in a human cadaver model of a surgical neck fracture. Ten fresh-frozen human cadaver shoulders were thawed, dissected free of soft tissue attachments, and analyzed with dual energy x-ray absorptiometry to establish bone mineral density. Osteotomies were fixed with figure-of-eight wire alone and figure-of-eight wire supplemented with intramedullary Enders rods. Intramedullary Enders rods improved the mean maximum load by 1.5 times (p < 0.05). No statistically significant correlation was found between mean maximum load and bone mineral density. PMID- 9356931 TI - Shoulder muscle moment arms during horizontal flexion and elevation. AB - The instantaneous muscle moment arms of 10 shoulder muscles including the three portions of the deltoid and the rotator cuff and scapulohumeral muscle groups during four specified glenohumeral motions were calculated. Moment arm values were derived from a plot of tendon excursion versus glenohumeral joint rotation angle during horizontal flexion along the 90 degrees elevation plane and elevation in the sagittal, scapular, and coronal planes. The deltoid had the largest moment arm in elevation. The anterior deltoid has a larger moment arm in the anterior planes, whereas the midportion is greater in the scapular and coronal planes. The muscles with the largest depressor (adductor) moment arms are the pectoralis major, latissimus dorsi, and teres major. Contrary to the findings of other investigators, the supraspinatus and infraspinatus have a smaller potential elevation torque in the scapular plane than has been previously reported. Furthermore the subscapularis may potentially be a more important elevator in the scapular plane than either the supraspinatus or infraspinatus, especially in the latter phases of motion. The pectoralis major has the largest horizontal flexion moment arm with the humerus elevated 90 degrees, whereas the posterior deltoid and infraspinatus have the largest horizontal extension moment arms in this plane. PMID- 9356932 TI - Evaluation of a proprioception pathway in patients with stable and unstable shoulders with somatosensory cortical evoked potentials. AB - Histologic studies have documented the presence of mechanoreceptors in the glenohumeral ligaments, capsule, and labrum; however, direct evidence of an intact afferent electrical pathway originating in structures in the shoulder is lacking. Because somatosensory cortical evoked potentials are transmitted by way of the dorsal columns of the spinal cord and carry proprioceptive information, this technique can be easily applied to evaluate the potential proprioceptive function of various intraarticular structures for shoulder stability. Patients have somatosensory cortical evoked potentials monitored while undergoing shoulder arthroscopy. The inferior glenohumeral ligament, middle glenohumeral ligament, subscapularis tendon, biceps tendon, supraspinatus rotator cuff capsule, glenoid labrum, and humeral head were evaluated. The intraarticular structures were stimulated with a monopolar electrode probe inserted through the anterior portal, and the evoked potentials were recorded with scalp electrodes. Generated wave forms were recorded and evaluated by measuring the peak-to-peak amplitude and latency. Three groups of patients with shoulder complications were studied: (1) no intraarticular pathologic condition and stable, (2) anterior instability with a Bankart lesion, and (3) anterior instability with a loose capsule. The articular cartilage of the humeral head generated no wave form in any subject. All other intraarticular structures generated consistent wave forms. No statistically significant difference was seen among the three groups when both amplitude and latency for the intraarticular structures were compared. PMID- 9356933 TI - Reduction of triceps muscle force after shortening of the distal humerus: a computational model. AB - Bone deficiency resulting in shortening of the distal humerus may occur after fractures, treatment of nonunions, or revision of total elbow arthroplasty. A biomechanical model of the triceps muscle and tendon was used to investigate the effect of distal humeral shortening on triceps force production. The analysis indicated that shortening of the distal humerus primarily influences the media head of the triceps, which contributes most to elbow extension strength in extended elbow positions. In a posture of 30 degrees elbow flexion, shortening the distal humerus by 1, 2, and 3 cm reduced the extension strength by 17%, 40%, and 63%, respectively. At 90 degrees of flexion, strength was reduced by 11%, 15%, and 21%, respectively. This result suggests that shortening the humerus by 1 cm may be well tolerated, but shortening by 2 or more cm may cause a significant reduction in triceps force. This would be particularly important in patients requiring terminal extension strength for weight bearing. PMID- 9356934 TI - Hemiarthroplasty for glenohumeral osteoarthritis: results correlated to degree of glenoid wear. AB - Thirty patients (31 shoulders) were retrospectively reviewed after hemiarthroplasty for glenohumeral osteoarthritis. Ten shoulders had primary osteoarthritis, and 21 shoulders had secondary osteoarthritis. Glenoid surface wear was evaluated and classified as either type I, concentric, (15 shoulders) or type II, nonconcentric, (16 shoulders). Postoperative results were reviewed with the American Shoulder and Elbow Surgeons' evaluation form, Neer classification, and the Constant score. Overall, 23 (74%) shoulders achieved satisfactory results, and 8 (26%) shoulders had unsatisfactory results. Results were similar in the primary and secondary osteoarthritis groups. Outcome correlated most significantly with the status of posterior glenoid wear. Patients with concentric, type I glenoids achieved 86% satisfactory results, whereas patients with nonconcentric, type II glenoids had only 63% satisfactory results. Although pain relief was similar in both groups, the unsatisfactory results were attributed to loss of forward elevation and external rotation in patients with type II glenoids. On the basis of these results hemiarthroplasty can be an effective treatment for both primary and secondary arthritis but should be reserved for patients with a concentric glenoid, which affords a better fulcrum for glenohumeral motion. PMID- 9356935 TI - Suprascapular nerve entrapment caused by supraglenoid cyst compression. AB - Twenty-two cases of suprascapular nerve entrapment caused by supraglenoid cyst compression were reviewed. Pain and weakness were the presenting symptoms in 14 shoulders and pain alone in 8. Twenty of the cysts were diagnosed by magnetic resonance imaging, and two were confirmed at surgical exploration. Electromyography of 20 shoulders was positive for neurologic involvement for both the infraspinatus and supraspinatus in 4 cases, for the infraspinatus only in 12, and negative in 4. Sixteen shoulders were treated by open excision, arthroscopy, or both. Superior labral lesions were diagnosed in 11 of 12 patients who underwent arthroscopy. At follow-up 10 of the patients who underwent surgery had complete resolution of symptoms, 5 had occasional pain or weakness, and 1 recurrence required a second surgery. Of six patients treated without surgery, two improved and four had no change. Supraglenoid ganglion cysts are common and can easily be diagnosed by magnetic resonance imaging. For patients with symptoms arthroscopy with repair of the superior labral lesion and either arthroscopic debridement or direct open decompression and excision of the cyst is recommended. PMID- 9356936 TI - Arthroscopic-assisted rotator cuff repair: patient selection and treatment outcome. AB - Over a 4-year period 24 patients out of 376 who required a rotator cuff repair were selected for arthroscopic-assisted rotator cuff repair. Preoperative selection criteria were refractory pain in the setting of good range of motion and strength (after an impingement test), absence of radiographic superior humeral head translation, and magnetic resonance imaging evidence of minimally retracted tear without rotator cuff muscle atrophy. Intraoperative selection criteria were the findings of an avulsion-type tear configuration with good tendon quality and absence of subscapularis tendon involvement. Based on these intraoperative criteria, 7 of the 24 patients were converted to an open approach to mobilize retracted and friable tendon tissue in a complex tear configuration. The remaining 17 patients underwent a transosseous arthroscopic-assisted rotator cuff repair with an average postoperative follow-up of 23 months. Evaluation by an independent therapist determined the postoperative American Shoulder and Elbow Surgeons Shoulder Function Index of 96 +/- 3 for the operative shoulder. The Functional Rating Scores for Activities of Daily Living and Sports Activity Score were 89% +/- 10% and 87% +/- 12%, respectively. Instrumented isometric strength for abduction and external rotation strength in the operated shoulder were 94% +/ 20% and 93% +/- 20%, respectively, compared with the contralateral unoperated side. Five of eight patients who performed overhead sports returned to a premorbid level of performance, and 14 of 15 patients available for follow-up believed that their result was excellent. We conclude that through careful selection one can identify patients optimally suited for arthroscopic-assisted rotator cuff repair, but some may have to be converted to an open end approach because of the quality of the tendon tissue and configuration of the tear requiring soft tissue releases. PMID- 9356937 TI - Strain of the anterior band of the inferior glenohumeral ligament during capsule failure. AB - Efficacious surgical treatment of glenohumeral instability requires a combination of anterior band origin repair and capsuloligamentous plication. The purpose of this article was to determine anterior band of the inferior glenohumeral ligament stretching at the time of glenohumeral failure. Thirteen fresh-frozen cadaver glenohumeral joints were thawed and dissected of soft tissue except for the capsuloligamentous structures. Testing was performed with a material testing system device, simulating the anterior instability apprehension position of the shoulder with 90 degrees of shoulder abduction and the humerus externally rotated until the bicipital groove was aligned with the supraglenoid tubercle. The length of each anterior band of the inferior glenohumeral ligament was obtained, and a variable reluctance transducer was applied to the anterior band midsubstance. Tensile testing at a strain rate of 100%/sec ensued until complete capsular failure occurred. Mid-substance strain of the anterior band of the inferior glenohumeral ligament at the time of capsular failure averaged 7.23% +/- 2.25% (mean +/- SD) with a range of 3.68% to 10.68%. Load to failure was 712.9 +/- 238.2 N (range 363.6 to 1136.9 N). All of the glenohumeral capsules failed at the glenoid origin, simulating a Bankart lesion, except for one that failed at the humeral insertion. When the intact capsuloligamentous tissue of the glenohumeral joint is tensile-tested in the apprehension position, there is only slight anterior band strain and failure occurring, predominantly at the glenoid insertion site. This has implications for the success of surgical procedures designed for acute repair of Bankart lesions. PMID- 9356938 TI - Posterior fracture-dislocation of the shoulder: treatment with acute osteochondral grafting. PMID- 9356939 TI - Pulmonary embolism caused by thrombosis of the axillary vein after shoulder arthroplasty. PMID- 9356940 TI - Oblique stress fracture of the olecranon in baseball pitchers. PMID- 9356941 TI - Destruction of contrast microbubbles during ultrasound imaging at conventional power output. AB - Inhomogenous opacification of cardiac chambers has been frequently observed after intravenous administration of long-persisting echocardiographic contrast agents. We observed this phenomenon to be most pronounced at high acoustic powers with incomplete opacification of the left ventricular apex and left ventricular outflow tract. Reducing the acoustic energy to which the contrast was exposed by decreasing transmit power or intermittently suspending insonification resulted in homogenous opacification of the entire left ventricular cavity. We systematically examined the effect of varying insonification power on the persistence of three investigational ultrasound contrast agents in both in vitro and in vivo models. We found an inverse relationship between the insonifying power and the persistence of the contrast agents. Contrast intensity decay could be reduced either by decreasing exposure to ultrasound by minimizing the transmit power of the system or by intermittently suspending ultrasound generation (triggering). Minimization of ultrasound contrast exposure to ultrasound energy thus improves echocardiographic contrast duration and homogeneity. PMID- 9356942 TI - Smaller intravenous perfluorocarbon-containing microbubbles produce greater myocardial contrast with intermittent harmonic imaging and better delineation of risk area during acute myocardial ischemia. AB - The purpose of this article was to determine whether applying negative or positive pressure to perfluorocarbon-containing microbubbles before intravenous injection would improve the myocardial contrast when using newer imaging techniques such as harmonic and intermittent imaging. Perfluorocarbon-containing microbubbles were exposed to sustained negative or positive pressure before intravenous injection in 10 dogs. Microbubble size distribution and concentration were measured after each exposure. Peak myocardial videointensity with intermittent harmonic imaging with each sample was compared. Microbubbles exposed to -200 mm Hg pressure before intravenous injection produced both the highest concentration of microbubbles and greater numbers of microbubbles less than 4 microns. Peak myocardial videointensity did not correlate with microbubble concentration or size but did correlate with the absolute number of microbubbles < 4 microns (mean r value 0.76, range 0.61 to 0.90). Risk area was best visualized with perfluorocarbon-containing microbubble samples containing the smallest microbubbles. We conclude that the myocardial contrast observed with perfluorocarbon-containing microbubbles can be enhanced by applying negative pressure before injection. The mechanism for this improved contrast appears to be related to creation of smaller microbubbles. PMID- 9356943 TI - Safety and efficacy of QW7437, a new fluorocarbon-based echocardiographic contrast agent. AB - The aim of this study was to evaluate the safety and efficacy of QW7437, a new fluorocarbon-based transpulmonary myocardial echocardiographic contrast agent. QW7437 is an anionically charged 2% dodecafluoropentane emulsion molecule, similar to EchoGen, a contrast agent previously shown to be efficacious in providing myocardial opacification by means of venous injection. This new agent has theoretical potential to provide greater safety and efficacy as a result of (1) reduced adherence to the negatively charged vascular endothelium and (2) reduced microbubble coalescence. Myocardial contrast echocardiography was performed in 10 dogs to evaluate the safety and efficacy of this agent. QW7437 (0.05 ml/kg) was injected as an intravenous bolus during intermittent harmonic epicardial imaging. Hemodynamic variables including heart rate, blood pressure, pulmonary artery pressure, pulmonary capillary wedge pressure, cardiac output, and arterial blood gases were determined at baseline and serially after contrast administration. Left ventricular fractional area shortening the regional myocardial blood flow at rest and during hyperemia (adenosine 140 micrograms/kg/min) were measured before and after contrast echocardiography. QW7437 provided dense myocardial opacification which persisted for more than 2 minutes in all subjects. This prolonged contrast effect did not result in significant changes in any hemodynamic variables, left ventricular function, or myocardial blood flow. Future studies should address the potential of this agent for human use. PMID- 9356944 TI - Global and regional atrial function after rapid atrial pacing: an echo Doppler study. AB - We examined diastolic transmitral, left atrial appendage, and pulmonary venous flow pulsed-wave Doppler velocities in five dogs sedated with Valium and ketamine both before and 1 week after rapid (400 beats/min) atrial pacing with transesophageal echocardiography. Left ventricular systolic function was assessed with long-axis fractional area shortening with the use of transesophageal echocardiography. Transmitral and left atrial appendage atrial systolic and pulmonary vein systolic velocities decreased significantly. The transmitral and left atrial appendage early-to-late velocity ratios increased significantly and the pulmonary venous systolic-to-diastolic velocity ratio decreased significantly after rapid atrial pacing. Thus, 1 week of rapid atrial pacing results in impaired global and regional atrial pump function and enhanced conduit function of the left atrium with preservation of left ventricular systolic function. These findings may have implications for anticoagulation therapy in patients with atrial arrhythmia. PMID- 9356945 TI - The value of dobutamine stress echocardiography for the detection of coronary artery disease in women. AB - To determine whether there were any gender-based differences in the detection of coronary artery disease by dobutamine stress echocardiography, we examined 288 patients (187 men and 101 women) who underwent coronary angiography within 8 weeks of dobutamine stress testing. Abnormal test results were indicated by let ventricular wall motion abnormalities at rest, which did not improve or worsen, or inducible wall motion abnormalities in two or more segments with dobutamine. Overall, dobutamine stress echocardiography showed a high sensitivity, specificity, and accuracy in both men and women: 85%, 96%, and 88% anx 90%, 79%, and 86%, respectively. The sensitivity in detecting significant coronary artery disease in our population was not influenced by gender. However, the sensitivity of the test was influenced by the extent and location of coronary disease and the pattern of left ventricular, hypertrophy. The sensitivity was 80% in patients with single-vessel disease, whereas the sensitivity was 91% in patients with multivessel disease. In addition, patients with single-vessel disease had lower sensitivity when the abnormality was located in the left circumflex coronary artery territory (59% versus 86% in the left anterior descending and right coronary territories). Our data indicated that there is no gender-based difference in the sensitivity and specificity of dobutamine stress echocardiography in detecting coronary artery disease and that the limitations of the test should be attributed to the extent and location of coronary disease. PMID- 9356946 TI - Measure the gap! A proposed simplified approach for measuring the descent of the base of the left ventricle. AB - The evaluation of left ventricular systolic function represents a common request for an echocardiographic examination. Several parameters have been proposed to quantitate left ventricular systolic function. The descent of the base of the left ventricle is founded on sound principle but currently requires multiple, time-consuming steps to derive during an echocardiographic examination. This article has reviewed the basic tenet of the descent of the base, reviewed the current required echocardiographic measurements needed to obtain a descent of the base value and has proposed a simplified approach by measuring the gap seen during the routine calculation of left ventricular end-systolic volume, end diastolic volume, and ejection fraction by the recommended method of discs. PMID- 9356947 TI - Echocardiographic estimation of left ventricular cavity area with a newly developed automated contour tracking method. AB - Development of an automated contour tracking method provides detection and tracking of the endocardial boundary using the energy minimization method without tracing a region of interest. The purpose of this study was to compare the automated contour tracking method and manually drawn methods for the measurement of left ventricular cavity areas and fractional area change. Apical four-chamber view was visualized and recorded for off-line analysis in 11 patients by means of two-dimensional echocardiography. The automated contour tracking method automatically traces the endocardial border from the recorded images and calculates left ventricular cavity areas (end-diastole and end-systole) and fractional area change. In the same images selected as end-diastole and end systole in the automated contour tracking method, left ventricular endocardial border was manually traced to calculate left ventricular cavity areas and fractional area change. Both methods were compared by linear regression analysis for the measurement of cavity areas and fractional area change. Left ventricular areas measured by the automated contour tracking method showed an excellent correlation with those by the manual method (end-diastole: r = 0.99, y = 0.83x + 2.6; standard error of the estimate = 1.5 cm2; end-systole: r = 0.99, y = 0.96x 0.8, standard error of the estimate = 1.2 cm2). The mean differences between the automated contour tracking and manual methods were -3.1 +/- 5.1 cm2 and -1.6 +/- 2.4 cm2 at end-diastole and end-systole, respectively. Fractional area change determined by the automated contour tracking method correlated well with that by the manual method (r = 0.95, y = 1.17x -6.5, standard error of the estimate = 3.4%). The mean difference between the automated contour tracking and manual methods was -0.8% +/- 7.1%. In conclusion, a newly developed automated contour tracking method correlates highly with the manual method for the estimation of left ventricular cavity areas and fractional area change in high-quality images. This suggests that this new technique may be useful in the automated quantitation of left ventricular function in patients with high-quality images with no dropout and no intercavity artifact or structure. PMID- 9356948 TI - Quantitative three-dimensional echocardiography by rapid imaging from multiple transthoracic windows: in vitro validation and initial in vivo studies. AB - Three-dimensional echocardiography has demonstrated superiority over two dimensional techniques in the determination of left ventricular mass and volumes. We describe a technique based on a magnetic tracking system which provides rapid three-dimensional image acquisition from multiple acoustic windows. Interactive three-dimensional border tracking and reconstruction with a piecewise smooth subdivision model accurately reproduced phantom volume (calculated volume = 1.00 true volume - 0.6 ml, r = 1.000, standard error of the estimate = 1.3 ml), in vitro heart volume (calculated volume = 1.02 true volume - 1.3 ml, r = 1.000, standard error of the estimate = 0.4 ml), in vitro heart mass (calculated mass = 0.98 true mass + 1.4 gm, r = 0.998, standard error of the estimate = 2.5 gm), and in vivo stroke volume (calculated stroke volume = 1.18 Doppler stroke volume - 17.9 ml, r = 0.990, standard error of the estimate = 2.8 ml). The three dimensional in vivo data sets, which include views from three acoustic windows, were acquired in less than 90 seconds. We conclude that this method of three dimensional echocardiographic data acquisition and analysis overcomes limitations inherent in currently available systems. PMID- 9356949 TI - Transthoracic three-dimensional echocardiographic volumetry of distorted left ventricles using rotational scanning. AB - The purpose of this study was to evaluate the relation of transthoracic three- and two-dimensional echocardiographic left ventricular volumetry to cineventriculographic volumetry. Twenty-five patients with distorted left ventricles were included in the study. To demonstrate the impact of acquiring data by rotational scanning, we performed three- and two-dimensional echocardiography in 36 latex ventricles with data acquisition in different areas of the ultrasound sectors. Interobserver and intraobserver variability were calculated to test for reproducibility. The three-dimensional imaging system consisted of a rotation motor device, a transthoracic 2.5 MHz transducer, a conventional ultrasound unit, and a work-station (TomTec) which provides data acquisition, post-processing, and two- or three-dimensional visualization of digitized data. The transducer moved automatically at 2-degree increments with data acquisition at each tomographic level. The mean investigation time for three dimensional echocardiography was 21 +/- 6 minutes. In the central near field of the transducer, differences from true volumes in latex ventricles were remarkably smaller for three-dimensional compared with two-dimensional echocardiography (root mean square percent error: three-dimensional echocardiography = 5.3% versus two-dimensional echocardiography = 14.6%). In three-dimensional echocardiography, there was considerable overestimation of volumes in the lateral far field (root mean square percent error = 13.2%) of the ultrasound sector. Differences between two-dimensional echocardiographic human left ventricular volumes and cineventriculography increased with larger volumes. In three-dimensional echocardiography the differences remained constant. Interobserver and intraobserver variability is reduced nearly twofold by three-dimensional echocardiography. Three-dimensional echocardiographic volumetry provides fewer discrepancies to cineventriculography and lesser variability than two-dimensional echocardiography. With the use of rotational scanning, the ventricle has to be positioned in the central near field of the transducer. PMID- 9356950 TI - Freehand three-dimensional echocardiography for determination of left ventricular volume and mass in patients with abnormal ventricles: comparison with magnetic resonance imaging. AB - OBJECTIVE: The objective of this study was to validate the freehand three dimensional echocardiographic method in patients with abnormal ventricular geometry compared with two-dimensional echocardiography using magnetic resonance imaging as a standard. BACKGROUND: Two-dimensional echocardiographic methods for estimating left ventricular volume and mass in clinical use today are limited by inaccuracies and variations caused by use of geometric assumptions and errors in image plane positioning. Freehand three-dimensional echocardiography with operator guidance by a "line of intersection" display eliminates these assumptions and errors. This method of volume and mass computation has been validated as highly accurate and reproducible in healthy subjects. METHODS: Left ventricular end-systolic and end-diastolic volumes and myocardial mass were determined by freehand three-dimensional echocardiography, by conventional two dimensional echocardiography using the apical biplane summation of discs method (volume) and the truncated ellipsoid method (mass), by M-mode echocardiography using the Penn method (mass), and by magnetic resonance imaging in 30 patients selected only for the presence of an abnormal ventricle. Results were compared by means of linear regression and the Bland-Altman method of analysis. RESULTS: There was excellent correlation, low bias, and low variability between three dimensional echocardiography and magnetic resonance imaging for end-diastolic volume (r = 0.90, standard error of the estimate = 31.8 ml, bias = -28.4 ml), end systolic volume (r = 0.93, standard error of the estimate = 24.1 ml, bias = -13.1 ml), and mass (r = 0.90, standard error of the estimate = 27.3 gm, bias = -22.6 ml). Two-dimensional echocardiography was less accurate and more variable as follows: end-diastolic volume (r = 0.70, standard error of the estimate = 39.8 ml, bias = -33.5 ml), end-systolic volume (r = 0.78, standard error of the estimate = 31.2 ml, bias = -26.7 ml), and mass (r = 0.80, standard error of the estimate = 37.3 gm, bias = 28.9 ml). M-mode echocardiography mass determination (Penn method) was least accurate and most variable (r = 0.075, standard error of the estimate = 78.3 gm, bias = 78.3 gm). CONCLUSIONS: Freehand three-dimensional echocardiography is a method of high accuracy and low variability for computing left ventricular volume and mass in clinical patients with abnormal ventricles. It is superior to conventional one- and two-dimensional echocardiography. The improvement achieved is attributed to elimination of geometric assumptions and image plane positioning errors and additional sampling of the ventricle. PMID- 9356951 TI - Ruptured aneurysm of the sinus of Valsalva into the left ventricle: a case report and review of the literature. AB - This report describes a case of right coronary sinus of Valsalva aneurysm which ruptured into the left ventricle. The diagnosis was made with two-dimensional transthoracic echocardiography which showed an abnormal structure extending from the aortic root into the left ventricle adjacent to the interventricular septum. Subsequent examinations with transesophageal echocardiography and aortic root angiography and surgical findings confirmed the diagnosis of transthoracic echocardiography. The patient underwent aortic valve replacement. At follow-up 12 months later, the patient was without symptoms and repeated echocardiographic examinations showed no recurrence. PMID- 9356952 TI - Pseudoaneurysm of the left ventricular outflow tract with reentry into the ascending aorta: an iatrogenic left ventricular ascending aortic fistula. AB - We report a patient with an iatrogenic pseudoaneurysm of the left ventricular outflow tract with reentry into the ascending aorta above the level of a prosthetic aortic valve. This pathology has not been previously described and was well demonstrated by transthoracic echocardiography. PMID- 9356953 TI - The tensor apparatus in double-orifice mitral valve: interpretation of echocardiographic findings. AB - We describe a rare case of double-orifice mitral valve discovered with echocardiography. The tensor apparatus composed of four papillary muscles and anomalous attachment of chordae tendineae represents the main findings. Two supernumerary muscles in combination with the altered chordal insertion on the central portion of the anterior mitral leaflet are responsible for the V-shaped ("seagull wing") and spectacle-like features of the mitral valve in the short axis view. These altogether allow precise identification of this congenital malformation. PMID- 9356955 TI - Resolution of vegetations with anticoagulation after myocardial infarction in primary antiphospholipid syndrome. AB - We report here a case of primary antiphospholipid syndrome with all three clinical features with acute myocardial infarction. Echocardiography showed large vegetations at both mitral valve leaflets. Laboratory evaluation showed presence of antiphospholipid antibodies. High-intensity anticoagulation was begun, and repeat echocardiographic study in 4 months showed disappearance of the mitral valve vegetations. PMID- 9356954 TI - Detection of a small left atrial myxoma: value and limitations of four imaging modalities. AB - Coronary angiography in a 52-year-old woman with angina-type chest pain showed tumor circulation with feeding arteries arising from the circumflex artery. Transthoracic echocardiography and magnetic resonance imaging both failed to show any intracardiac masses. A sessile mass measuring 1.5 cm in diameter attached to the atrial septum was readily detected by transesophageal echocardiography. Histologic analysis confirmed the tumor to be a myxoma. Coronary angiography may provide the first clue to the presence of a small myxoma. Transesophageal echocardiography is the imaging modality of choice for further evaluation. PMID- 9356956 TI - Adherence issues in HIV therapeutics. Introduction: the situation. PMID- 9356957 TI - Adherence as a particular issue with protease inhibitors. AB - Suboptimal therapeutic doses of HIV protease inhibitors lead to the emergence of drug resistance and reduced drug efficacy. It is imperative, then, that patients and clinicians alike are fully educated about the importance of patients taking all the pills in these new, admittedly complex, antiretroviral regimens. With protease inhibitors especially, missing doses and/or taking drug holidays or partial doses will mean the rapid emergence of HIV isolates that are resistant to these drugs. PMID- 9356958 TI - Antiretroviral therapy: factors associated with adherence. AB - Clinical effectiveness of the new highly active antiretroviral therapies depends in large part on patients' ability to adhere to demanding medication regimens because the suboptimal drug levels associated with nonadherence are, in turn, associated with the development of antiretroviral resistance. However, definitions of adherence are inconsistent, and the concept is difficult to measure. Adherence to medical and health regimens is the outcome of a dynamic process of human behavior and interaction. Factors influencing this process include characteristics of the regimen, the provider, the patient, and society. PMID- 9356959 TI - The case manager's role in adherence. AB - The nurse as case manager has a responsibility to help patients become fully informed before making decisions about specific antiretroviral therapy regimens. In addition, the case manager can help patients assess their current situation with regard to the possibility of taking and adhering to complex antiretroviral regimens. For the patient who is currently taking antiretroviral therapy, the case manager is the point person who oversees continuity of care and ensures communication among all of the patient's health care providers. PMID- 9356960 TI - HIV drug adherence: special situations. AB - Among the highly diverse population of persons living with HIV/AIDS are individuals with particularly challenging life circumstances that can be called "special situations." Substance abuse and homelessness are examples of special situations that require additional consideration when attempting to determine the appropriateness of prescribing complex antiretroviral regimens. When individual cases are examined in the context of relevant models of care and the principles of those models applied, such clinical decisions can be made with the patient. Withholding protease inhibitors from an entire population group, it is argued, is the epitome of practicing bad medicine. PMID- 9356961 TI - Adherence to prescribed HIV-1 protease inhibitors in the home setting. AB - The issue of adherence to any therapy prescribed by a health care provider is multidimensional and includes personal challenges that exist in the daily life of any client, as well as the individual practices and attitudes of the health care provider. In addition, since most health is delivered in congregate settings such as the hospital, clinic, or office, it is important to consider those factors that may be present in a client's home that can impede compliant behavior. This article reviews challenges to compliance that should be considered when developing a care plan for clients, focusing on variables that may be present in the home setting. PMID- 9356963 TI - Will a presidential public apology be needed for HIV/AIDS care in the future? PMID- 9356964 TI - Correlates of spiritual well-being in terminally ill persons with AIDS and terminally ill persons with cancer. AB - In an effort to determine if terminally ill patients with AIDS had greater religious and spiritual care needs than other terminally ill patient populations, particularly those with cancer, a study was conducted in a community-based hospice in the southeast. The purpose of the study was to compare the perceptions of spiritual well-being, loneliness, social support, health hardiness, pain, and functional status among terminally ill clients with cancer and terminally ill clients with AIDS in a hospice setting and to examine predictors of spiritual well-being in a hospice population. A sample of 55 hospice patients completed the Correlates of Spiritual Well-Being Scale (COSWEB), which includes a demographic data sheet and instruments to measure spiritual well-being, loneliness, health hardiness, social support, functional status, and pain. Patients with AIDS reported significantly lower spiritual well-being than did patients with cancer and other chronic, terminal illnesses. Patients with AIDS also reported significantly greater loneliness than other patient populations. The number of social supports for patients with AIDS was significantly lower than for cancer patients and other groups; moreover, patients with AIDS were significantly more dissatisfied with their supports than other patient groups. The best predictors of spiritual well-being in this study were social support and loneliness, which explained 47% of the variance in spiritual well-being. The results of this study suggest differences between specific groups of hospice patients. Patients with AIDS may be less spiritually well than other terminally ill patient populations due to decreased support systems, dissatisfaction with supports, greater feelings of loneliness, younger ages on entry to hospice, fewer family supports, lack of recognized long-term relationships, and related issues such as homophobia, perceived rejection by religious denominations, unstable living environments, economic disadvantages, and less time to process life events/meaning. Findings in this study and similar future studies can better enable health care providers to allocate time and resources to various terminally ill patient populations to achieve higher quality care outcomes in general and greater spiritual well-being in particular. PMID- 9356962 TI - HIV therapeutics: confronting adherence. PMID- 9356965 TI - Reconstruction case management. PMID- 9356966 TI - Development of an HIV educational needs assessment tool. AB - The learning needs of persons with HIV/AIDS continue to grow more complex as new treatments are developed and HIV disease shifts from being a terminal to a chronic illness. Clinicians could use a simple tool to help them to focus on what clients think is important to know about living with HIV/AIDS. This article describes the development of the HIV Educational Needs Assessment Tool (HENAT). HIV+ persons (N = 363) who were receiving health care in a variety of ambulatory and institutional settings participated in this research during 1990 and 1993 1994. Factor analysis was used to shorten the instrument into a form that could be used in any nurse/client interaction. Principal axis factor extraction and varimax rotation deleted 16 items. The remaining 34 items were grouped into six factors: Treatments, Entitlements, Relationships, Preventing Infections, Social Support, and Working. Cronbach's alphas were computed and results ranged from .70 to .88. Two month test-retest correlations for a subset of participants (n = 195) ranged from .54 to .67. HENAT can be used (a) to examine differences in perception of learning needs between clients with HIV disease and their health care providers, (b) to measure shifts in learning needs over time, (c) to relate HIV-specific learning needs to length of time living with HIV disease, (d) to give clients an opportunity to assess their learning needs, and (e) as part of a larger intervention study that evaluates the effectiveness of patient education. PMID- 9356967 TI - The role of self-esteem in safer sexual practices. AB - Self-esteem appears in the literature as a variable that influences the practice of risky sexual behaviors. It is often assumed that higher levels of self-esteem are associated with safer sexual behaviors, especially those that prevent the spread of HIV. The research literature was reviewed to examine the relationship between self-esteem and the practice of safer sexual behaviors. Research indicates that higher levels of self-esteem are found in adolescents who practice risky sexual behaviors and have more sexual partners. Research and clinical implications are discussed. PMID- 9356968 TI - Nutritional issues and HIV/AIDS: assessment and treatment strategies. AB - Ongoing nutritional assessment is vital in the care of adults and children with HIV infection. Nutritional deficiency is a common manifestation of HIV disease and results from a variety of factors. Nurses caring for individuals with HIV infection in a variety of settings can be instrumental in identifying nutritional deficits, linking patients with appropriate medical/nutritional support services, and assuring that appropriate nutritional interventions are implemented. This article summarizes etiologies of HIV-associated nutritional deficiencies, reviews important components of nutrition assessment (including nutrition-related side effects of approved medications commonly used in HIV disease), provides an overview of common nutritional problems and interventions, and lists some available nutritional resources. PMID- 9356969 TI - Ritonavir (Norvir). PMID- 9356970 TI - Heterosexual transmission of HIV in women. PMID- 9356971 TI - A sage brain, a sturdy skeleton, and a funny bone: a longevity lesson from Madame Calment. PMID- 9356972 TI - News from the Society for the Advancement of Women's Health Research. PMID- 9356973 TI - Setting the research agenda for the 21st century. PMID- 9356974 TI - Gender is important: lessons in autoimmunity. PMID- 9356975 TI - Selective estrogen receptor modulators and postmenopausal women's health. AB - Selective estrogen receptor modulators represent an alternative approach to the use of estrogen replacement therapy or hormone replacement therapy for decreasing postmenopausal bone loss, as well as for reducing the incidence of serious cardiovascular disease in this population. Of particular interest is raloxifene, a benzothiophene compound, which binds with high affinity to the estrogen receptor and produces effects similar to estrogen on the skeleton and cardiovascular system but behaves as a complete estrogen antagonist in the uterus and the breast. The pharmacologic profile of raloxifene, a discussion of a possible mechanism of action, and the potential role of this drug in women's postmenopausal health are the subjects of this review. PMID- 9356976 TI - Medical complications of anorexia nervosa. AB - Anorexia nervosa is often characterized by progressive deterioration in many different organ systems. Most medical complications are the result of starvation and can be reversed with a well-planned refeeding program. While some of the complications of anorexia nervosa are predictable physiologic adaptations to the self-imposed starvation, many others are potentially life threatening. It is therefore incumbent upon all primary care physicians to become familiar with this disorder, because it is increasing in incidence and is commonly burdened by substantial chronicity and recidivism. PMID- 9356977 TI - Symptoms of post-traumatic stress disorder in abused women in a primary care setting. AB - Abuse is a major source of trauma to women, and post-traumatic stress disorder (PTSD) results from exposure to extreme trauma. To describe the relationship between symptoms of PTSD and severity of abuse, an ethnically stratified cohort of 131 abused women in a primary care setting was interviewed. Symptoms of PTSD, both intrusion (i.e., trouble falling asleep, strong waves of feelings about the abuse) and avoidance (i.e., trying not to think or talk about the abuse, staying away from reminders of the abuse), were significantly (p < 0.01) correlated to severity of abuse, regardless of ethnicity. When asked about childhood physical or sexual abuse, women reporting physical abuse had significantly (p < 0.05) higher intrusion scores, whereas those reporting sexual abuse had significantly (p < 0.004) higher avoidance scores. Sixty-five percent of the women reported dreams, flashbacks, or terror attacks and had significantly (p < 0.001) higher mean results on both intrusion and avoidance. The need to offer abused women information about the connection between severity of abuse and symptoms of PTSD is discussed. We recommend that clinicians ask all abused women about dreams, flashbacks, or terror attacks to assess for further symptoms of PTSD. PMID- 9356979 TI - The decision-making process for the treatment of abnormal uterine bleeding. AB - This pilot study was conducted to investigate the treatment decision-making process of patients and physicians for abnormal uterine bleeding (AUB). Frequently, women with AUB are referred for hysterectomy without diagnostic workup, alternative therapeutic management, or patient input (i.e., patient treatment preferences). Variations in treatment strategies used for patients may be related to a number of factors external to the patient's underlying disease. However, little is known about which factors are most influential or about the extent to which they influence physicians' and patients' decisions. We prospectively followed the management and treatment of 52 women with complaints of AUB and examined differences in treatment among these patients. Extensive previsit interviews were conducted with these women to identify each patient's symptoms, health status, functional status, and preferences for and expectations of treatment. We then conducted telephone interviews within a week of the visit and again 9-12 months later to determine the treatment plan, patient level of participation in and satisfaction with the treatment, symptoms, and functional status. Overall, our findings suggest that patients want to be involved in making treatment decisions and that when women were presented with alternatives to hysterectomy, many chose alternative medical therapy or other surgical procedures. In addition, women reported that these alternative treatments produced significant improvement in symptom intensity and functioning. Increased patient participation in decision making enhanced patient satisfaction. These findings suggest that hysterectomy rates may be decreased by offering women alternative treatments and by finding ways to increase women's participation in their treatment decisions. PMID- 9356978 TI - Thyroid function and perimenopausal lipid and weight changes: the Thyroid Study in Healthy Women (TSH-W). AB - We designed a prospective observational trial to study the relationship of thyroid function to cholesterol and weight changes at menopause. Subjects were participants in the ongoing Healthy Women Study, a prospective study of cardiovascular risk factor change through menopause. Healthy premenopausal women were recruited from a random sample of licensed drivers in selected ZIP codes of Allegheny County, Pennsylvania. Participants had to be 42-50 years of age, have menstruated within the last 3 months, not have had surgical menopause, have diastolic blood pressure < 100 mm Hg, and not be taking medications (including insulin, estrogen, lipid-lowering drugs, or thyroid or antihypertensive medications) at the baseline examination. The substudy included three groups of women who were premenopausal at baseline and were categorized according to change noted at follow-up regarding menopausal status and use of hormone replacement therapy (HRT). The groups comprised 95 women who remained premenopausal, 96 postmenopausal women not on HRT, and 61 postmenopausal women using HRT. The main outcome measures were baseline and follow-up measurements for serum levels of thyroid-stimulating hormone (TSH), thyroid peroxidase, and thyroglobulin, as well as serum cholesterol, total high-density lipoprotein (HDL) cholesterol, triglycerides, and calculated low-density lipoprotein (LDL) cholesterol, height, and weight. Covariates included cigarette smoking and alcohol intake. The prevalence of thyroid antibodies in this healthy population was high at both time points (range 27%-31%) and did not differ by menopausal status. The presence of thyroid antibodies was associated with increased TSH concentration. Women with antibodies at both time points had lower levels of total and LDL cholesterol compared with those with no antibodies, significant only for those women who remained premenopausal during the follow-up period. Thyroid function during menopause in this healthy population is unlikely to account for the observed changes in levels of serum lipoprotein and body weight. The presence of thyroid antibodies may be associated with lower total and LDL cholesterol, possibly through an underlying inflammatory disorder. PMID- 9356980 TI - Use of mental health services by HIV-infected women. AB - In this study, the first to examine the use of mental health services among a large cohort of HIV-positive women, our objective was to quantify the use of such services by infected women enrolled at one site of the HIV Epidemiology Research Study and to describe the factors associated with such use. One hundred sixty seven HIV-seropositive women and 67 seronegative women were interviewed between March and November 1995 during their semiannual study visit. Women were asked to report visits they had made to mental health counselors (psychiatrists, social workers, or psychologists) or HIV support groups and any psychiatric hospitalizations during the previous 6 months. About half the seropositive women had a history of injection drug use, were Caucasian, and lived with children, three quarters were insured, and one third were church members. A minority of seropositive women (38%) sought at least one outpatient mental health visit, and 4% had been hospitalized during the previous 6 months. Being a member of a church, having a high school education, and being Caucasian were significantly associated (p < 0.05) with seeing a mental health counselor. Among HIV-positive women who reported at least one visit, their CD4 cell count was the only factor associated with the number of mental health visits. Only 13% of women had attended an HIV support group. Among seronegative women, 27% had at least one outpatient mental health visit during the preceding 6 months. The use of mental health services by women with HIV has economic, not just therapeutic, implications for all HIV service delivery systems. PMID- 9356981 TI - The quality of information on women's health on the Internet. AB - The World Wide Web is an excellent information resource for professionals and the public to gain clinical knowledge. Internet technology has changed the way we perceive and present data because of the speed with which complex data manipulations are now possible and the vast quantities of data involved. The need for current, comprehensive data gave rise to a prevailing tradition of clear, factual, and impartial information resources on the Internet. Most arguments in favor of regulating and restricting the information that is accessible to patients underestimate the power of the new health care consumers and fail to distinguish between quality of information and quality of knowledge. PMID- 9356982 TI - Search on urinary incontinence in elderly females. PMID- 9356983 TI - C-X-C chemokine receptor desensitization mediated through ligand-enhanced receptor phosphorylation on serine residues. PMID- 9356984 TI - Generation of monoclonal antibodies to chemokine receptors. PMID- 9356985 TI - Chemokine receptors in developing human brain. PMID- 9356986 TI - Expression of chemokine receptors in insect cells using baculovirus vectors. PMID- 9356988 TI - Molecular approaches to identifying ligand binding and signaling domains of C-C chemokine receptors. AB - The construction of chimeric receptors provides a useful starting point for the identification of ligand-binding and G-protein-coupling sites on chemokine receptors. Correlation of the binding and signaling properties of a set of complementary receptor chimeras is a powerful approach for probing structure function relationships. Further molecular resolution can subsequently be achieved by site-directed mutagenesis and/or alanine scanning. PMID- 9356987 TI - Chimeric chemokine receptors for analysis of structure-function relationships. PMID- 9356989 TI - Characterization of functional activity of chemokine receptors using the Cytosensor Microphysiometer. PMID- 9356990 TI - Calcium flux assay of chemokine receptor expression in Xenopus oocytes. PMID- 9356991 TI - Cell-cell fusion assay to study role of chemokine receptors in human immunodeficiency virus type 1 entry. PMID- 9356992 TI - Iodination of chemokines for use in receptor binding analysis. PMID- 9356993 TI - Expression of chemokine receptors by endothelial cells: detection by intravital microscopy using chemokine-coated fluorescent microspheres. PMID- 9356994 TI - Neutralization of interleukin-8 in in vivo models of lung and pleural injury. PMID- 9356995 TI - Murine experimental autoimmune encephalomyelitis: a model of immune-mediated inflammation and multiple sclerosis. PMID- 9356997 TI - Role of chemokines in antibacterial host defense. PMID- 9356996 TI - In vitro and in vivo systems to assess role of C-X-C chemokines in regulation of angiogenesis. PMID- 9356998 TI - Animal models of asthma: role of chemokines. AB - In studies of disease processes, increasing knowledge leads to an increased awareness of the complexity of the underlying mechanisms. The intense research activity in the chemokine field has made this acutely manifest. Numerous chemokines have been discovered through the use of (1) bioassay of in vitro cell culture supernatants and in vivo exudates from animal models of inflammation and (2) molecular biology techniques. Any one chemokine can often be produced by a number of different cell types and exert its effects on different target cells. This has been interpreted by some as implying a high degree of redundancy. Although this is understandable, in disease processes parallel and sequential mechanisms are possible, and potentially important therapeutic targets have emerged. There is compelling evidence from animal and clinical studies that eosinophils are important effector cells in asthma, but this relationship is as yet unproven in the human disease. Two possible targets to prevent eosinophil recruitment to the lung are IL-5 and its receptor, which are important in several aspects of eosinophil biology, and eotaxin and its receptor, CCR3. The eotaxin receptor is particularly attractive as a target as it is expressed in high numbers on eosinophils, but not other leukocytes, and appears to be the major detector of the eosinophil for eotaxin and other chemokines such as MCP-4. Eotaxin and CCR3 knockout mice are being developed, and animal models will continue to be invaluable when antagonists are available. In the shape of receptor antagonists, the chemokine field may yet provide the final proof of concept for the long-established eosinophil theory of asthma in humans. PMID- 9356999 TI - Identification and structural characterization of chemokines in lesional skin material of patients with inflammatory skin disease. PMID- 9357002 TI - Adenylate cyclase assays to measure chemokine receptor function. PMID- 9357001 TI - G-protein activation by chemokines. PMID- 9357000 TI - Calcium mobilization assays. PMID- 9357003 TI - Analysis of signal transduction following lymphocyte activation by chemokines. PMID- 9357004 TI - Calcium mobilization and phosphoinositide turnover as measure of chemokine receptor function in lymphocytes. PMID- 9357005 TI - Effect of differentiation on the leucine enkephalin-degrading soluble enzymes released by the K562(S) cell line. AB - Leu-enkephalin hydrolysis kinetics were measured in the presence of soluble supernatants obtained from cultures of the K562(S) leukaemic cell line. Under these conditions, the substrate is degraded with formation of two distinct patterns of the hydrolysis by-products: in one pattern, similar amounts of Tyr and Tyr-Gly are formed; in the other, only Tyr-Gly can be measured. Kinetic data suggest that soluble proteolyses are released by these cells, and that either dipeptidylaminopeptidases alone, or both aminopeptidases and dipeptidylaminopeptidases are involved in substrate hydrolysis. This alternation of hydrolysis patterns appears consistent with existing data on the heterogeneity of K562 cells. In contrast with these results, chromatographic separation of the soluble enzymes indicates the release of all three classes of proteolyses known to hydrolyze enkephalins: aminopeptidases, dipeptidylaminopeptidases and dipeptidylcarboxypeptidases. In cells induced to differentiate by treatment with butyric acid, substrate hydrolysis is increased, and the pattern of the enzymes released is modified. In these cells, variations in both total proteolytic activity, and ratio between the three enzyme classes mentioned above are only minor, while the ratio between the different enzyme species within each class is greatly modified. Data obtained suggest that the expression of soluble enzymes is modified by differentiation. These data may also be interpreted as stressing the role of competition in controlling substrate hydrolysis by the multiple enzymes co-released by K562(S) cells. PMID- 9357006 TI - Intrinsic innervation of the chicken lower digestive tract. AB - We have studied the different components of the enteric nervous system in the rectum and cloaca of the chicken by means of histochemical and immunohistochemical techniques. We found cholinergic neuronal bodies as well as nervous fibers, which constitute part of the Meissner and Auerbach plexuses. We also observed plentiful catecholaminergic fibers in both plexuses, though there were no catecholaminergic neuronal bodies. With respect to the Vasoactive Intestinal Peptide (VIP) and substance P (SP) positive peptidergic innervation, only positive fibers were found, which were less abundant than in the other zones of the gastrointestinal tract. The optic microscopy results were confirmed by electron microscopy. PMID- 9357007 TI - Effects of beta-amyloid-(25-35) peptides on radioligand binding to excitatory amino acid receptors and voltage-dependent calcium channels: evidence for a selective affinity for the glutamate and glycine recognition sites of the NMDA receptor. AB - The neurotoxic fragment corresponding to residues 25-35 of the beta-amyloid (A beta) peptide [A beta-(25-35)] has been shown to exert effects on (+)-[3H]5 methyl-10, 11-dihydro-5H-dibenzo[a,d]-cyclohepten-5,10-imine maleate ([3H]MK-801) binding to the cation channel of the N-methyl-D-aspartate (NMDA) receptor. In the present study, we investigated whether the amidated and carboxylic acid C terminated forms of A beta-(25-35) [A beta-(25-35-NH2) and A beta-(25-35-COOH), respectively] exert effects on other excitatory amino acid receptor and cation channel types in rat cortical membranes. Both A beta-(25-35-NH2) and A beta-(25 35-COOH) gave statistically significant dose-dependent inhibitions of [3H]glutamate and [3H]glycine binding to the agonist recognition sites of the NMDA receptor. Ten microM A beta-(25-35-NH2) and A beta-(25-35-COOH) gave 25% and 20% inhibitions of [3H]glutamate binding and 75% and 70% inhibitions of [3H]glycine binding, respectively. A beta-(25-35-NH2), but not A beta-(25-35 COOH), gave a small (ca. 17% at 10 microM) statistically significant increase of [3H]amino-3-hydroxy-5-methylisoxazole-4-propionate ([3H]AMPA) binding. [3H]kainate binding was not significantly affected by either peptide. Similarly, neither peptide affected either the maximal level or EC50 value for calcium stimulation of [3H]nitrendipine binding. It is concluded that A beta-(25-35) shows slight affinity for the agonist recognition sites of the NMDA receptor, but not for other excitatory amino acid receptor types or for L-type voltage dependent calcium channels. PMID- 9357008 TI - Manganese decreases glutamate uptake in cultured astrocytes. AB - Recent data have shown an accumulation of manganese in the basal ganglia in patients with chronic hepatic encephalopathy (HE). Astrocytes and ammonia are critically involved in the pathogenesis of HE, and we have recently demonstrated that ammonia decreases glutamate uptake in cultured astrocytes. Since failure by astrocytes to take up glutamate may represent an important pathogenetic mechanism in HE, we, therefore, examined the effect of manganese on glutamate transport in these cells. Treatment of cultured astrocytes with 100 microM manganese for 2 days resulted in a 54% decrease in the uptake of D-aspartate, a nonmetabolizable analogue of glutamate. Kinetic analysis revealed a 28% decline in Vmax, with no change in the K(m). Treatment of cultures with 5 mM NH4 Cl inhibited D-aspartate uptake by 21%, and a combination of 5 mM NH4Cl with 100 microM manganese produced an additive effect on uptake inhibition. These results suggest a pathogenetic role for manganese in HE, possibly involving glutamate transport. PMID- 9357009 TI - Neurotrophic activity of organosulfur compounds having a thioallyl group on cultured rat hippocampal neurons. AB - Several organosulfur compounds found in garlic extract promoted the survival of rat hippocampal neurons in vitro. From the analysis of structure-activity relationship, thioallyl group in these compounds is essential for the manifestation of neurotrophic activity. S-Allyl-L-cysteine (SAC), one of the organosulfur compounds having thioallyl group in garlic extract, also promoted the axonal branching of cultured neurons. These results suggest that thioallyl compounds make a unique group of neurotrophic factors. PMID- 9357010 TI - N-terminal arginylation of sciatic nerve and brain proteins following injury. AB - N-terminal protein arginylation has been demonstrated in vitro and in situ and has been reported to increase following injury to sciatic nerves of rats. The present study attempts to demonstrate these reactions in vivo by applying [3H]Arg to the cut end of sciatic nerves in anesthetized rats and assaying for N-terminal arginylation using Edman chemistry and acid precipitation of labeled proteins in the proximal nerve segment. No evidence was found for arginylation in an aqueous soluble fraction. However, N-terminal arginylation was detected in a urea soluble fraction at 2 hours after nerve crush. The data show that arginylation of rat sciatic nerve proteins occurs in vivo and suggest that the arginylated proteins formed an aqueous insoluble/urea soluble aggregate after arginylation. In other experiments, rat brains were injured and assayed for arginylation in vitro to test the hypothesis that injury causes an up-regulation of these reactions. Results showed an activation of the reaction at 2 hours post crush and indicate that increases in N-terminal arginylation are likely to be a general response to injury in nervous tissue. PMID- 9357012 TI - Interaction of divalent metal ions with Zn(2+)-glycerophosphocholine cholinephosphodiesterase from ox brain. AB - The effect of divalent metal ions on the activity of glycerophosphocholine cholinephosphodiesterse from ox brain was examined. Zn(2+)- and Co(2+) glycerophosphocholine cholinephosphodiesterases were prepared from the exposure of apoenzyme to Zn2+ and Co2+, respectively, and the properties of two metallo phosphodiesterases were compared to those of native phosphodiesterase. Although two metallo-enzymes were similar in expressing Km value, optimum pH or sensitivity to Cu2+, they differed in the susceptibility to the inhibition by thiocholine or tellurite; while Co(2+)-phosphodiesterase was more sensitive to tellurites, Zn(2+)-phosphodiesterase was more susceptible to inhibition by thiocholine. In addition, Zn(2+)-phosphodiesterase was more thermo-stable than Co2+ enzyme. Separately, when properties of native phosphodiesterase were compared to those of each metallo-phosphodiesterase, native phosphodiesterase was found to be quite similar to Zn(2+)-phosphodiesterase in many respects. Even in thermo-stability, native enzyme resembled Zn(2+)-phosphodiesterase rather than Co(2+)-enzyme. Consistent with this, the stability of native phosphodiesterase was maintained in the presence of Zn2+, but not Co2+, Mn2+ was also as effective as Zn2+ in the stabilization of the enzyme. Noteworthy, the native enzyme was found to be inhibited competitively by Cu2+ with a Ki value of 20 microM, and its inhibitory action was antagonized effectively by Zn2+ or Co2+. Also, choline, another competitive inhibitor of the enzyme, appeared to antagonize the inhibitory action of Cu2+. Taken together, it is suggested that there may be multiple binding sites for divalent metal ions in the molecule of glycerophosphocholine cholinephosphodiesterase. PMID- 9357011 TI - Effect of osmolality and myo-inositol deprivation on the transport properties of myo-inositol in primary astrocyte cultures. AB - myo-Inositol uptake measured in primary astrocyte cultures was saturable in the presence of Na+ with a Km of 13-18 microM and a Vmax of 9.4 nmoles/mg protein/hour in myo-inositol-fed cells, indicating a high affinity transport system. In myo-inositol-deprived cells, Km was about 53 microM with a Vmax of 13.2 nmoles/mg protein/hour. Decreasing osmolality decreased the Vmax to about 1.9 nmoles/mg protein/hour whereas increasing osmolality increased Vmax about 5 fold, while Kms were essentially unchanged in myo-inositol fed cells. In cells deprived of myo-inositol, Vmax decreased in hypotonic medium and increased in hypertonic medium almost 10-fold, but with more than a doubling of the Km regardless of the osmolality. Glucose (25 mM) inhibited myo-inositol uptake 51% whereas the other hexoses used inhibited uptake much less. Our findings indicate that myo-inositol uptake in astrocytes occurs through an efficient carrier mediated Na(+)-dependent co-transport system that is different from that of glucose and its kinetic properties are affected by myo-inositol availability and osmotic stress. PMID- 9357013 TI - Excitatory sulfur-containing amino acid-induced release of [3H]GABA from rat olfactory bulb. AB - The effect of L-cysteine sulfinic acid (CSA) and L-homocysteic acid (HCA) on the release of tritiated gamma-amino butyric acid ([3H]GABA), from the external plexiform layer (EPL) of the rat olfactory bulb, was compared with that of glutamate. These amino acids induced release of GABA was strongly inhibited by the glutamate uptake blocker, pyrrolidine-2,4-dicarboxylate (2,4,PDC) (50 microM), while it was not inhibited by the specific GABA uptake blockers nipecotic acid (0.5 mM) or NO-711 (5 microM). Only the HCA induced GABA release was 60% inhibited by beta-alanine (0.5 mM), a glial GABA uptake blocker and 78% by the NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid (AP-5) (100 microM). The non-NMDA receptor antagonists 6-cyano-2,3-dihydroxy-7-nitro quinoxaline (CNQX) up to 500 microM had no effect on HCA or CSA stimulated GABA release. These results bring evidence for an excitatory role of HCA and CSA together with glutamate on GABAergic neuronal or glial elements, in the olfactory bulb. This role could be mediated through the reversal of the glutamate or/and the glial GABA transporter and through the activation of a NMDA type receptor. PMID- 9357014 TI - M1 muscarinic receptor stimulation decreases aspartate release in the rat neostriatum. AB - This study investigates the effects of different muscarinic receptor agonists on extracellular glutamate and aspartate concentrations in the rat neostriatum. In vivo intracerebral perfusions were undertaken in the conscious rat using a concentric push-pull cannulae system. Amino acid concentrations in samples were determined by HPLC with fluorometric detection. The intrastriatal perfusion of arecoline, a M1-M2 muscarinic receptor agonist, produced a significant decrease in extracellular [ASP] (45% of decrease) but not in extracellular [GLU]. These effects were blocked by scopolamine, a M1-M2 muscarinic receptor antagonist. McN A-343, a M1 muscarinic receptor agonist, but not the M2 muscarinic receptor agonist, oxotremorine, produced a significant decrease in extracellular [ASP] (40% of decrease) but not in extracellular [GLU]. The effects of McN-A-343 on extracellular [ASP] were blocked by pirenzepine, a M1 muscarinic receptor antagonist. These results suggest that the decrease in extracellular [ASP] could be mediated, at least in part, by M1 muscarinic receptor activation in the rat neostriatum. PMID- 9357016 TI - Variable deposition of amyloid beta-protein (A beta) with the carboxy-terminus that ends at residue valine40 (A beta 40) in the cerebral cortex of patients with Alzheimer's disease: a double-labeling immunohistochemical study with antibodies specific for A beta 40 and the A beta that ends at residues alanine42/threonine43 (A beta 42). AB - Amyloid beta-protein (A beta) deposits in the cerebral cortices of patients with Alzheimer's disease (AD) were investigated immunohistochemically to determine their carboxy terminal sequences. Antibodies specific for A beta terminating at residue valine40 (A beta 40) and at residues alanine42/threonine43 (A beta 42) were used. Virtually all parenchymal A beta deposits were positive for A beta 42. Many of these deposits were also partially or completely labeled for A beta 40. The degree of A beta 40 labeling varied from area to area within a given brain and from AD case to AD case. In contrast to parenchymal deposits, A beta 40 labeled essentially all the vascular deposits which constitute amyloid angiopathy (AA), with A beta 42 occurring variably in some of these deposits. Occasional AA was found, however, in which A beta 42 predominated or was exclusively deposited. Such a diversity of A beta species, both in brain parenchyma and in AA, suggests that multiple C-terminal processing mechanisms occur in the cell types responsible for these deposits. PMID- 9357015 TI - Effects of 4-aminopyridine on extracellular concentrations of glutamate in striatum of the freely moving rat. AB - 4-aminopyridine (4-AP) is a voltage-sensitive K(+)-channel blocker extensively used in in vitro experiments as a depolarizing agent for the release of glutamate (GLU). This research investigated whether 4-AP could be used in in vivo experiments using microdialysis. For that, the effects of 4-AP on the extracellular concentrations of glutamate (GLU), glutamine (GLN), taurine (TAU) and citrulline (CIT) in striatum of the freely moving rat were investigated. The effects of 4-AP were compared with those produced by perfusion with a high K+ (100 mM) medium. Intrastriatal perfusion with 4-AP (1, 5 and 10 mM) produced no effects on extracellular [GLU], [TAU] and [CIT], but decreased extracellular [GLN]. Perfusion with a high K+ (100 mM) medium increased extracellular [GLU] and [TAU], decreased extracellular [GLN], and had no effects on [CIT]. To test whether the lack of effects of 4-AP on extracellular [GLU] was due to GLU uptake mechanisms, 4-AP was perfused after a previous inhibition of GLU uptake with L trans-pyrrolidine-2,4-dicarboxylic acid (PDC). Under the effects of PDC (1 mM), 4 AP (1 mM) had no effects on extracellular [GLU], [TAU] and [CIT], but decreased extracellular [GLN]. These results show that 4-AP decreased extracellular [GLN] but failed to produce a significant release of GLU in striatum of the freely moving rat. Thus, 4-AP can not be used as a depolarizing agent for stimulating the release of GLU in in vivo studies using microdialysis. PMID- 9357017 TI - Effect of gamma-decanolactone on glutamate binding in the rat cerebral cortex. AB - Epilepsy is one of the most common neurological disorders. Even though antiepileptic drugs can afford a reasonably satisfactory treatment for 80% of diagnosed patients, chronic intractable epilepsy still affects a significant number of people and more effective and less harmful antiepileptic drugs are needed. Previous studies have shown that gamma-decanolactone has dose-dependent sedative effects, including hypnotic, anticonvulsant and hypothermic properties in mice. The present study reports an inhibitory effect of gamma-decanolactone on glutamate binding (96.8% with 5 mM) in rat cortex membranes. The non competitive nature of glutamate binding inhibition as a neurochemical correlate of the anticonvulsant activity of gamma-decanolactone may be a relevant mode of action for further drug development. PMID- 9357019 TI - Effect of nitric oxide donors on GABA uptake by rat brain synaptosomes. AB - The effect of nitric oxide donors and L-arginine on the uptake of GABA was studied in synaptosomes purified from rat brain. The neurotransmitter uptake was significantly reduced by S-nitrosoacetylpenicillamine and by sodium nitroprusside, although in this case to a lesser extent. A slight inhibitory effect was found preincubating rat brain synaptosomes with 1 mM L-arginine as well. The S-nitrosoacetylpenicillamine effect gradually disappeared with decomposition of the substance by exposure to light. The nitric oxide effect appears to be mainly due to a decrease in the V for synaptosomal GABA uptake and seems to be related to a partial collapse of nerve endings ionic gradients. Functionally, it could result over time in a reduced availability of GABA at the synapses involved. PMID- 9357018 TI - Tau-like proteins in the nervous system of goldfish. AB - This report describes the presence of a group of tau-like proteins (TLPs) in goldfish central nervous system. The TLPs were immunoreactive with antibodies that recognized the carboxy-terminal domain of mammalian tau, but not with antibodies that recognized the amino-terminus. The TLPs of goldfish exhibited the basic properties of tau proteins including neuronal specificity, structural heterogeneity, heat stability and the ability to co-assemble with tubulin. We propose that TLPs may represent a precursor of tau, that share the microtubule binding domain and the carboxy-terminal domain with mammalian tau proteins. In contrast the amino-terminus of the TLPs is much shorter and may represent a more variable domain of tau proteins. PMID- 9357021 TI - The immune system in eating disorders: an overview. AB - Eating disorders, such as anorexia nervosa and bulimia nervosa, are becoming more and more common in our society. Although they are psychiatric illnesses, there are many factors involved, including abnormal food behavior. Nutrients play an important role in the development and functionality of immunocompetent cells. An impaired immunocompetence has been shown to be an important causal factor in the increased susceptibility of malnourished individuals to infectious disease. Therefore, studies on the immune system are of great interest when assessing the extent to which the nutritional status of these patients could be affected. However, the literature in this field is controversial, and the mechanisms are not yet completely defined, although some hypotheses try to clarify the disturbances caused in the organism under these bizarre circumstances. In spite of the fact that the immune system is altered by distorted food behaviors, such as in eating disorders, the awareness of characteristics of other systems involved, and therefore altered, by these pathologies would be very helpful for understanding the mechanisms triggered in these syndromes. In fact, the interactions among the immune and other systems in eating disorders are beginning to be studied. Finally, the main goals are to limit the evolution of these illnesses through early diagnosis, and to devise a long-lasting, definitive cure for these patients through appropriate therapy. PMID- 9357020 TI - Iron-induced inhibition of Na+, K(+)-ATPase and Na+/Ca2+ exchanger in synaptosomes: protection by the pyridoindole stobadine. AB - The effect of oxidative stress, induced by Fe(2+)-EDTA system, on Na+,K(+) ATPase, Na+/CA2+ exchanger and membrane fluidity of synaptosomes was investigated. Synaptosomes isolated from gerbil whole forebrain were incubated in the presence of 200 microM FeSO4-EDTA per mg of protein at 37 degrees C for 30 min. The oxidative insult reduced Na+,K(+)-ATPase activity by 50.7 +/- 5.0% and Na+/Ca2+ exchanger activity measured in potassium and choline media by 47.1 +/- 7.2% and 46.7 +/- 8.6%, respectively. Membrane fluidity was also significantly reduced as observed with the 1,6-diphenyl-1,3,5-hexatriene probe. Stobadine, a pyridoindole derivative, prevented the decrease in membrane fluidity and in Na+/Ca2+ exchanger activity. The Na+,K(+)-ATPase activity was only partially protected by this lipid antioxidant, indicating a more complex mechanism of inhibition of this protein. The results of the present study suggest that the Na+/Ca2+ exchanger and the Na+,K(+)-ATPase are involved in oxidation stress mediated disturbances of intracellular ion homeostasis and may contribute to cell injury. PMID- 9357022 TI - Glutamine-enhanced bacterial killing by neutrophils from postoperative patients. AB - Neutrophils play an important role in host defense by phagocytosing and destroying invading bacteria. A recent investigation revealed that glutamine (Gln) augmented the in vitro bactericidal activity of neutrophils from burn patients. However, it is unclear whether Gln enhances the function of neutrophils in postoperative patients. This study was designed to investigate the effect of Gln on the in vitro Escherichia coli-killing activity of neutrophils from postoperative patients. Nine randomly selected patients were included in this study. On the morning of the first postoperative day, blood was drawn and neutrophils were isolated. Eight healthy volunteers served as controls. E. coli was opsonized with pooled normal serum. Neutrophils (5 x 10(6)), together with opsonized E. coli (5 x 10(5)), were incubated for 2 h at 37 degrees C in Hanks' balanced salt solution supplemented with 0, 100, 500, or 1000 nmol/mL of Gln. The bactericidal function of neutrophils was determined by counting the number of viable bacteria. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL 8, and granulocyte elastase levels in the cell culture supernatant were measured. Plasma C-reactive protein (CRP), cortisol, and amino acids were also analyzed. The plasma concentration of Gln was significantly lower in the postoperative patients than in the controls. Following culture with patient neutrophils, the number of viable E. coli decreased by 26% as the in vitro Gln concentration was increased from 500 to 1000 nmol/mL (P < 0.01). We defined the Gln 1000/Gln 500 ratio of the number of viable bacteria as the number of viable E. coli at an in vitro Gln concentration of 1000 nmol/mL divided by the number of viable E. coli at an in vitro Gln concentration of 500 nmol/mL. A positive correlation was thus demonstrated between the plasma Gln level and the Gln 1000/Gln 500 ratio of the number of viable bacteria in the patients (r = 0.69, P = 0.04). This finding indicated that as plasma Gln fell, there was an enhancement of neutrophil E. coli killing activity by neutrophils in in vitro tests when the Gln concentration was increased from 500 to 1000 nmol/mL. Gln supplementation caused no appreciable changes in TNF-alpha, IL-1 beta, IL-8, or granulocyte elastase levels in cell culture supernatants. A negative correlation was recognized between the patient plasma Gln level and the Gln 1000/Gln 500 ratio of the cell culture supernatant IL-8 level (r = -0.73, P = 0.025). In conclusion, Gln supplementation enhanced the in vitro bactericidal function of neutrophils from postoperative patients. PMID- 9357023 TI - A 10-year survey of nutritional support in a surgical ICU: 1986-1995. AB - Total parenteral nutrition (TPN) has long been considered the optimal nutrition technique in critically ill patients, but recently the use of enteral nutrition (EN) has increased. This study describes the evolution of the different nutritional support techniques in a surgical intensive care unit (ICU) in a university hospital, through (1) a global survey over 10 y assessing the evolution of the use of EN and TPN, and (2) a prospective study performed over 6 mo. Severity of illness and diagnostic categories were stable (n = 11,539 patients). From 1986 to 1990, the proportion of TPN administered increased from 10-25% of ICU days, decreasing to 10% thereafter. EN was used in about 5% of ICU days in 1986, and had increased to 30% of total ICU treatment days in 1995. The proportion of nutrients actually delivered to the patients was 75% with EN and 88% with TPN. Major changes in nutritional support have been observed since 1986. The frequency of nutritional support provided in general has increased to 40% of ICU treatment days. TPN has been largely overtaken by EN, with the risk of insufficient energy delivery, related to the difficulties of EN in the critically ill. These results reinforce the importance of continuous quality control by daily assessment of nutrient supply. PMID- 9357024 TI - Nutritional and immunologic evaluation of patients with gastric cancer before and after surgery. AB - The main objective was to evaluate a patient's immunologic and nutritional status as a prognostic indicator of morbidity and mortality in patients with gastric cancer. A prospective clinical study carried out at the National Cancer Institute in Bogota, Colombia. Our study group consisted of 40 patients with a diagnosis of gastric adenocarcinoma that was treated surgically. Blood samples were taken before and 5 d after surgery; mononuclear cell typing was done by flow cytometry allowing a bicolor analysis. Nutritional evaluation was obtained through measurement of albumin levels, average weight loss, and nutritional risk index (NRI). Half of the malignancies were localized to the middle and lower third of the stomach: stage I, 17.55%; stage II, 10%; stage III, 55%; and stage IV, 17.5%. Twenty subtotal gastrectomies, 11 total gastrectomies, 7 gastrojejunostomies, and 2 esophagogastrectomies with D1 and D2-D3 lymph node resection were performed. A postoperative morbidity of 22.5% and a mortality of 7.5% were observed. A preoperative cellular immunosuppression was identified, with a helper lymphocyte (CD4) to suppressor/cytotoxic lymphocyte (CD8) ratio of 1.38 normal value (NV > 1.5), which increased according to the stage of the disease. Patients who died presented with a significantly greater preoperative cellular immunosuppression than those who survived (P = 0.05). Postoperative mortality correlated significantly with hypoalbuminemia (P = 0.008). In those who died, weight loss was greater than in those who survived (P = 0.06). Patients with severe malnutrition had greater postoperative mortality according to the NRI. Severe preoperative cellular immunosuppression (CD4/CD8 < 1), hypoalbuminemia, weight loss, and severe NRI have a positive predictive value for mortality in patients with gastric cancer. PMID- 9357025 TI - Effects of MCT/LCT and LCT emulsions on plasma lipids and nitrogen retention in streptozotocin-induced diabetic rats receiving total parenteral nutrition. AB - This study was designed to investigate the effects of emulsions containing medium chain triacylglycerols (MCT) or long-chain triacylglycerols (LCT) on plasma lipids and nitrogen retention in diabetic rats receiving total parenteral nutrition (TPN). Diabetes was induced in rats by streptozotocin (STZ). Control and diabetic rats were divided into two TPN groups. The TPN groups received solutions at an energy level of 30 kcal/100 g body weight with 37.5% of the nonprotein energy provided as fat. All TPN solutions were isonitrogenous and identical in nutrient composition except for the fat emulsion, which was composed of LCT or MCT/LCT (1:1). The results showed that plasma triacylglycerol (TG), nonesterified fatty acids (NEFA), and beta-hydroxybutyrate levels were higher in diabetic rats compared with control rats, whereas plasma insulin levels and nitrogen retention were lower. Plasma glucose levels, TG, NEFA, and beta hydroxybutyrate levels were significantly decreased after TPN administration in diabetic groups. Plasma glucose and TG levels, however, remained higher in diabetic groups than in control groups. No difference in the concentrations of plasma TG, cholesterol, NEFA, beta-hydroxybutyrate or nitrogen retention were observed between the two diabetic groups. These results suggest that MCT/LCT infusion did not lead to hyperketonemia and hypercholesterolemia as compared with LCT infusion, and had no beneficial effect on nitrogen retention in rats with STZ induced diabetes under the present experimental conditions. PMID- 9357026 TI - The effect of alpha-adrenergic antagonism upon nitrogen loss during endotoxemia. AB - Sixty male Sprague-Dawley rats were randomized to receive parenteral nutrition (PN) only; PN plus continuous infusion of Escherichia coli 026:B6 lipopolysaccharide (PN + LPS) at 6 mg.kg-1.d-1; or PN plus LPS plus a continuous infusion of the alpha-adrenergic antagonist phentolamine (PN + LPS + PHEN) at 5 mg.kg-1.d-1 or 20 mg.kg-1.d-1 for 48 h. All animals received isocaloric, isonitrogenous PN. LPS significantly lowered nitrogen balance (mmol/48 h) from PN control; however, addition of PHEN substantially worsened nitrogen balance compared with LPS (14.2 +/- 3, 2.4 +/- 5.2, -1.6 +/- 4.5, -0.8 +/- 5.4, for the PN, PN + LPS, PN + LPS + PHEN5 and PN + LPS + PHEN20 groups, respectively; P < 0.0001). Urinary 3-methylhistidine/creatinine ratio (3-meH/creat) paralleled the nitrogen balance data (0.30 +/- 0.09, 0.45 +/- 0.12, 0.51 +/- 0.14, 0.60 +/- 0.12, respectively; P < 0.0001). The high-dose PHEN resulted in 82 +/- 9% blockade. To ascertain if any beneficial effect upon body protein loss is achieved during severe stress, 30 rats were given PN + LPS at 12 mg.kg-1.d-1 or PN + LPS12 + PHEN20. These data showed similar changes in nitrogen balance and 3 methylhistidine/creatinine with the use of PHEN during severe endotoxemia. alpha adrenergic antagonism with PHEN worsens body protein loss as measured by nitrogen balance and 3-methylhistidine/creatinine in PN-fed endotoxemic rats. PMID- 9357028 TI - Does intra-abdominal fluid increase the resting energy expenditure? AB - In patients with intra-abdominal fluid collection, caloric needs are based on an estimated dry weight. This is done because intra-abdominal fluid has been assumed to be metabolically inactive. One recent study of patients with slowly resolving ascites suggested otherwise. In our study, the effect of intra-abdominal fluid on the resting energy expenditure (REE) and apparent lean body mass was determined in 10 stable patients requiring peritoneal dialysis. For each subject, in both the empty and full state, we measured REE by indirect calorimetry, and body composition by the bioelectric impedance method. In the full state, the VCO2 was significantly increased (210 +/- 11 versus 197 +/- 9 mL/min, P < 0.02) compared with the empty state. This caused an increase in the calculated resting energy expenditure (1531 +/- 88 kcal/d empty versus 1593 +/- 94 kcal/d full, P < 0.05). The magnitude of increase in REE was similar to the expected calories derived from glucose absorbed out of the dialysate. Estimates of body fat, lean body mass, and total water also were not affected by the intra-abdominal fluid. We conclude that intra-abdominal fluid will not affect the measured REE and hence may be considered to be metabolically inactive. PMID- 9357027 TI - Benfluorex, a hypotriglyceridemic drug, reduces lipid peroxidation and alleviates adverse metabolic complications of copper deficiency. AB - The pathologies associated with copper deficiency in rats fed fructose may be induced, in part, by hypertriglyceridemia and lipid peroxidation. Reducing triacylglycerol levels in plasma may result in lowering lipid peroxidation, which in turn could ameliorate metabolic effects resulting from the combination of fructose feeding and copper deficiency. Benfluorex, a hypolipidemic factor able to reduce hypertriglyceridemia, was administered to weanling male rats fed either copper-deficient (0.6 microgram Cu/g) or adequate (6.0 micrograms Cu/g) diets containing fructose as the sole dietary carbohydrate. In copper-deficient rats, benfluorex (50 micrograms.kg-1.d-1) reduced plasma triacylglycerols from 45 to 31 mg/dL, reduced lipid peroxidation by approximately 50%, and prevented the enlargements of heart and liver size and the atrophy of the pancreas, and ameliorated anemia. It is suggested that lipid peroxidation associated with hypertriglyceridemia may be responsible for the pathologies induced by the combination of fructose consumption and copper deficiency. PMID- 9357029 TI - Nutrition of the fetus and premature infant. AB - Nutrition in the fetus and the premature infant is a rapidly changing field, not solely due to the acquisition of new knowledge but also because there have been major conceptual advances that have altered our approach to nutrition during early stages of development. This special report will highlight some of these conceptual advances in this area. PMID- 9357031 TI - Glutamine and infections. PMID- 9357030 TI - The clinical management of short bowel syndrome: steps to avoid parenteral nutrition. PMID- 9357032 TI - Nutrition in the ICU--from overfeeding to starvation. PMID- 9357035 TI - Lipid peroxidation and DNA damage. PMID- 9357034 TI - Novel candidate biomarkers for dietary chemoprevention of colon cancer. PMID- 9357033 TI - Vitamin D compounds as potential therapeutics for estrogen-independent breast cancer. PMID- 9357036 TI - Modulation of RAS expression in human malignant cells by dietary supplements. PMID- 9357037 TI - Adolescent diet and the risk of breast cancer in adulthood: a role for vitamin A? PMID- 9357039 TI - Cost-effectiveness of nutrition support teams. Are they necessary? PMID- 9357040 TI - Carpe diem: the Taiwanese health care reform. PMID- 9357038 TI - Changes in Medicare reimbursement policy may restrict nutrition therapy options. PMID- 9357041 TI - Present conditions and the future role of hospital dietitians in Japan. PMID- 9357042 TI - Filtration of parenteral nutrition admixtures: friend or foe. PMID- 9357043 TI - Incisional hernias after laparoscopy. AB - Laparoscopy, using a two-puncture technique, has been used for a variety of gynecologic indications for more than two decades. The procedure is considered safe and effective, although rare complications, such as incisional hernias, have been reported. In this review, the issue of postlaparoscopic incisional hernias is discussed in terms of incidence, predisposing factors, time of appearance, and preventive measures. With the evolvement of operative laparoscopy, larger trocars and cannulas have been introduced, increasing the incidence of postlaparoscopic incisional hernias. Awareness of the possibility of this complication will lead to the use of proper surgical techniques, as suggested, while knowledge of the postoperative signs and symptoms will lead to early diagnosis and prevention of sequelae. PMID- 9357044 TI - Complementary and alternative medicine. Part I: Clinical studies in obstetrics. AB - Studies in which complementary and alternative medicine (CAM) interventions were used in the care of obstetric patients were identified by searching The National Library of Medicine's electronic database. This paper is a selected review of both randomized and nonrandomized clinical trials. There are a number of CAM therapeutic practices that may have potential benefit for patients. Additional clinical research with appropriate scientific methodology is warranted to determine the merit and value of integrating some CAM modalities into conventional medical obstetric care. PMID- 9357045 TI - Complementary and alternative medicine. Part II: Clinical studies in gynecology. AB - This review describes randomized and nonrandomized clinical studies in which complementary and alternative interventions were used to treat gynecologic illness and disease. The articles were identified through the use of The National Library of Medicine's electronic database. Statistically significant clinical benefit could not be identified for most practices. However, additional scientific investigation with appropriate research methods would help clarify a number of provocative insights and suggestions of potential clinical effectiveness. PMID- 9357046 TI - Structure-activity relationships of FMRFamide-related peptides contracting Schistosoma mansoni muscle. AB - This study reports the potent myoactivity of flatworm FMRFamide-related peptides (FaRPs) on isolated muscle fibers of the human blood fluke, Schistosoma mansoni. The turbellarian peptides YIRFamide (EC50 4 eta M), GYIRFamide (EC50 1 eta M), and RYIRFamide (EC50 7 eta M), all induced muscle contraction more potently than the cestode FaRP GNFFRFamide (EC50 500 eta M). Using a series of synthetic analogs of the flatworm peptides YIRFamide, GYIRFamide and RYIRFamide, the structure-activity relationships of the muscle FaRP receptor were examined. With a few exceptions, each residue in YIRFamide is important in the maintenance of its myoactivity. Alanine scans resulted in peptides that were inactive (Ala1, Ala2, Ala3 and Ala4 YIRFamide; Ala4 and Ala5 RYIRFamide) or had much reduced potencies (Ala1, Ala2 and Ala3 RYIRFamide). Substitution of the N-terminal (Tyr1) residue of YIRFamide with the non-aromatic residues Thr or Arg produced analogs with greatly reduced potency. Replacement of the N-terminal Tyr with aromatic amino acids resulted in myoactive peptides (FIRFamide, EC50 100 eta M; WIRFamide, EC50 0.5 eta M). The activity of YIRFamide analogs which possessed a Leu2, Phe2 or Met2 residue (EC50's 10, 1 and 3 eta M, respectively) instead of Ile2 was not significantly altered, whereas, YVRFamide had a greatly reduced (EC50 200 eta M) activity. Replacement of the Phe4 with a Tyr4 (YIRYamide) also greatly lowered potency. Truncated analogs were either inactive (FRFamide, YRFamide, HRFamide, RFamide, Famide) or had very low potency (IRFamide and MRFamide), with the exception of nLRFamide (EC50 20 eta M). YIRF free acid was inactive. In summary, these data show the general structural requirements of this schistosome muscle FaRP receptor to be similar, but not identical, to those of previously characterized molluscan FaRP receptors. PMID- 9357047 TI - Neuropeptide FF-like immunoreactivity in human cerebrospinal fluid of chronic pain patients and healthy controls. AB - Neuropeptide FF (NPFF) is a neuropeptide with some antiopioid characters found in several mammalian species. In human brain it might be an important pain regulating peptide. Using a specific and sensitive radioimmunoassay we found a mean concentration of NPFF in human cerebrospinal fluid (CSF) of healthy volunteers of 1.6 +/- 1.1 pg/ml (n = 19) and in chronic pain (CPD) patients of 1.4 +/- 1.2 pg/ml (n = 16). The NPFF concentrations in CSF and plasma did not correlate. There was no difference in the NPFF concentrations in CSF and plasma between CPD patients and healthy controls. NPFF in CPD patients did not correlate significally with any pain characteristic. This study provides evidence for the presence of NPFF in human brain, but does not support the hypothesis that chronic pain is a consequence of elevated production of NPFF. PMID- 9357048 TI - afp-1: a gene encoding multiple transcripts of a new class of FMRFamide-like neuropeptides in the nematode Ascaris suum. AB - We have identified a gene, afp-1, that encodes a new subfamily of six FMRFamide like neuropeptides in the nematode Ascaris suum. The predicted peptides share the C-terminal sequence PGVLRF-NH2 but have different N-terminal extensions. We discuss possible functional roles of these different peptides based upon experiments with Ascaris as well as results from other organisms. Three of the peptides were previously isolated from extracts of A. suum (4) and three other are novel sequences. The translated product of afp-1 is a precursor protein containing two main halves: a C-terminal region containing a series of putative peptides separated by characteristic processing sites and a relatively hydrophobic N-terminal region with no obvious peptides. Although the overall structure of the translated product of afp-1 is similar to flp-1 from C. elegans (18), there is little evidence for homology between the two nematode neuropeptide genes. At least four different transcripts of afp-1 have been identified. These transcripts differ in their 3' and 5' untranslated regions, and one of the transcripts predicts a truncated precursor protein which contains only the C terminal peptide-containing region. PMID- 9357049 TI - Identification of three allatostatins and their cDNA from the mosquito Aedes aegypti. AB - Three allatostatins have been isolated from the mosquito Aedes aegypti. These peptides have the following structures: Ser-Pro-Lys-Tyr-Asn-Phc-Gly-Leu-amide, Leu-Pro-His-Tyr-Asn-Phe-Gly-Leu-amide, and Arg-Val-Tyr-Asp-Phe-Gly-Leu-amide. A cDNA encoding these peptides was isolated from an abdominal ganglia cDNA library and sequenced. It was found to encode two additional allatostatins: Ala-Ser-Ala Tyr-Arg-Tyr-His-Phe-Gly-Leu-amide and Leu-Pro-Asn-Arg-Tyr-Asn-Phe-Gly-Leu-amide. Northern analysis of whole mosquito mRNA revealed a single prepro-allatostatin message of around 3,000 bases. Identification of a partial prepro-allatostatin cDNA from a midgut cDNA library shows that the same gene is also expressed in the mosquito midgut. PMID- 9357050 TI - The NMDA receptor antagonist, NPC 12626, reduces the pronociceptive effects of orphanin FQ and kappa opiate antinociception in the land snail, Cepaea nemoralis. AB - The heptadecapeptide, orphanin FQ or nociceptin (Phe-Gly-Phe-Thr-Gly-Ala-Arg-Lys Ser-Ala-Arg-Lys-Leu-Ala-Asn-Gln), originally isolated from rat brain has been identified as an endogenous ligand for the orphan opioid-like receptor. Although orphanin FQ shares some sequence and structural homology with kappa-opioid peptides, it has been speculated to exert its effects through novel nonopioid mechanisms. Kappa opioids have also been suggested to have nonopioid actions in rodents involving the N-methyl-D-aspartate (NMDA) receptor. The present study examined the effects of the competitive NMDA antagonist, NPC 12626, on the antinociceptive effects of the specific kappa-opiate receptor agonist, U69,593, and the pronociceptive effects of orphanin FQ in an invertebrate system, the land snail, Cepaea nemoralis. NPC 12626 had no effect on the basal nociceptive sensitivity of snails, as measured by the latency of response to a thermal (40 degrees C) surface. As reported for rodents, NPC 12626 dose-dependently reduced U69,593-induced antinociception in a manner comparable to that produced by the specific kappa-opiate antagonist, nor-binaltorphimine, while slightly enhancing the antinociceptive effects of the predominately mu-opiate agonist, morphine. Similarly, NPC 12626 dose-dependently reduced the pronociceptive effects of orphanin FQ. These findings with the snail, Cepaea, indicate that NMDA systems/receptors are associated with the mediation of the nociceptive effects of both kappa opioids and orphanin FQ. They suggest an early evolutionary development and phylogenetic continuity of NMDA opioid and related neuropeptide interactions in the mediation of nociception. PMID- 9357052 TI - Acetyl salmon endorphin-like immunoreactivity in the ovary of two teleostean species: changes with environmental conditions. AB - The presence of salmon acetylated endorphin (acetyl sEP) in the ovary of seabream and sea bass was investigated through immunocytochemical and biochemical techniques in order to compare aquatic species with terrestrial ones. Endorphin like immunoreactivity was found in the cytoplasm of oogonia and similar immunostaining was present in the granulosa layer of mature follicles. In both pituitary and ovarian extracts of the two teleostean species, acetyl sEP-like immunoreactivity was distributed over three main peaks, the second one corresponding to the elution time of the reference synthetic peptide. Serial dilutions of HPLC fraction II of the ovaries of both fishes ran parallel with the standard curve obtained with reference peptide. The ovarian content of acetyl sEP, obtained by calculating the integrated area of the fraction II peak, indicates large and highly significant (p < 0.01) differences in the amount of peptide found in ovarian tissues of wild seabream in comparison with that of farmed fish. Increased peptide values in wild animals with respect to farmed fish were also found in the sea bass. These data indicate that not only the pituitary, but also the ovary is sensitive to environmental cues, and strongly suggest the role of opioid peptides in adaptation. PMID- 9357051 TI - Differential metabolism of dynorphins in substantia nigra, striatum, and hippocampus. AB - To map the proteolytic enzymes metabolizing dynorphins in brain structures, size exclusion chromatography linked to electrospray ionization mass spectrometry was used. Enzymes extracted from rat hippocampus, striatum, and substantia nigra were tested for their capability of converting dynorphin-related peptides. Dynorphin A was the most resistant to proteolytic conversion, whereas Big dynorphin and dynorphin B-29 were slowly converted to dynorphin A and dynorphins A and B, respectively. Dynorphin B and alpha-neoendorphin were the least resistant. Dynorphin B was rapidly converted to Leu-enkephalin in the striatum and hippocampus but to Leu-enkephalin-Arg6 in the substantia nigra. alpha Neoendorphin was converted to Leu-enkephalin in all tissues investigated. PMID- 9357053 TI - ACTH/CLIP immunoreactivity in the cat brain stem. AB - The distribution of adrenocorticotropin hormone/corticotropin-like intermediate lobe peptide was studied in the cat brain stem, using an indirect immunoperoxidase technique. No immunoreactive cell bodies were observed. However, a high density of immunoreactive fibers was found in the periaqueductal gray, the dorsal nucleus of the raphe, the locus coeruleus, and the marginal nucleus of the brachium conjunctivum. A moderate density was found in the central linear nucleus, the central tegmental field, the Kolliker-Fuse nucleus, the inferior central nucleus, and the postpyramidal nucleus of the raphe. A low density was found in the superior and inferior colliculi, the interpeduncular nucleus, the nucleus sagulum, the superior central nucleus, the cuneiform nucleus, the accessory dorsal tegmental nucleus, the nucleus of the solitary tract, the dorsal motor nucleus of the vagus, and the paralemniscal, magnocellular, gigantocellular, and lateral tegmental fields. Moreover, single immunoreactive fibers were observed in numerous nuclei of the cat brain stem. In comparison with previous studies carried out in the same region of the cat, as well as the rat and the human, our results point to a more widespread distribution of adrenocorticotropin hormone/corticotropin-like intermediate lobe peptide immunoreactive structures in the cat brain stem. This widespread distribution indicates that the peptide might be involved in several physiological functions of the cat brain stem. PMID- 9357054 TI - Angiotensinergic neurons physiologically inhibit prolactin, growth hormone, and thyroid-stimulating hormone, but not adrenocorticoptropic hormone, release in ovariectomized rats. AB - Angiotensin II (AII)-containing neurons with cell bodies in the rostral medial hypothalamus and axons project to the external layer of the median eminence, so that AII maybe released into the hypophyseal portal vessels for actions on the pituitary gland. Indeed, intrahypothalamic actions of the peptide on the release of hypothalamic hormones and direct actions on the pituitary have been reported. To determine the role of endogenously released AII in hypothalamic-pituitary hormone release, we have determined the effects of central immunoneutralization of AII upon the plasma concentrations of prolactin (PRL), growth hormone (GH), thyroid-stimulating hormone (TSH), and adrenocorticotropic hormone (ACTH). Specific antiserum directed against AII (AB-AII) or normal rabbit serum (NRS), as a control, was microinjected into third ventricular (3 V) cannulae of conscious, ovariectomized (OVX) rats. Immediately before and at various intervals after this procedure, blood samples were withdrawn through previously implanted external jugular catheters. Three hours after injection of the AB-AII, plasma PRL levels diverged from those of the NRS-injected animals and progressively increased from 4 to 24 h after administration of the antiserum. Results were similar with respect to plasma GH, except that the increase in the AB-AII animals above that in the NRS-injected controls from 4 to 6 h was not significant, but was highly significant on measurement 24 h after injection, at which time plasma GH was three times higher than in control rats. Similarly, following injection of AB AII, plasma TSH values did not diverge significantly from those of the NRS injected controls until 3 h after injection. From 3 to 5 h they remained constant and significantly elevated above values in the NRS-injected controls with a further nonsignificant increase at 6 h. At 24 h, there was no longer a difference between the values in both groups. In contrast to the significant elevations in plasma hormone levels observed with respect to PRL, GH, and TSH following injection of the antiserum, there was no change in plasma ACTH between the AB-AII injected and NRS-injected animals throughout the same period of observation. Previous results by others have shown that intraventricular injection of AII has a suppressive action on the release of PRL, GH, and TSH. Consequently, we believe that the antiserum is acting intrahypothalamically to block the action of AII within the hypothalamus, resulting in the elevation of the three hormones mentioned. Therefore, the AII neurons appear to have a physiologically significant suppressive action on the release of hypothalamic neurohormones controlling the release of PRL, GH, and TSH. In contrast, there apparently is no effect of intrahypothalamically released AII on the secretion of corticotropin releasing factors under these nonstress conditions. We cannot rule out an action of the antiserum at the pituitary level; however, in view of the fact that the actions of AII directly on the gland are to stimulate PRL, GH, TSH, and ACTH release, it appears that the antiserum was acting at the hypothalamic level. PMID- 9357055 TI - Role of brain angiotensin in thirst and sodium appetite of rats. AB - The role of brain angiotensin II (ANG II) in water, Na and food intake of rats was studied. Intracerebroventricular (i.c.v.) infusion (100 micrograms/h) of the non-peptide ANG II receptor antagonist losartan (type 1), but not PD123319 (type 2), completely blocked water intake caused by i.c.v. infusion of ANG II at 50 ng/h. Following food deprivation, food intake was reduced by PD123319 and associated water intake was decreased by losartan or PD123319. Neither water intake after water deprivation nor Na intake after Na depletion was altered by losartan or PD123319. In conclusion, evidence was consistent with a role for brain ANG II in both food and water intake after food deprivation but not in thirst subsequent to water deprivation or Na intake after Na depletion alone. PMID- 9357057 TI - Bombesin regulation of adrenocorticotropin release from ovine anterior pituitary cells. AB - Mammalian members of the bombesin-like peptide family (gastrin releasing peptides; GRP) have been localized in the ovine median eminence and in hypophysial-portal blood, suggesting a role in the regulation of anterior pituitary function. In this study we have shown that although bombesin cannot stimulate ACTH secretion alone, it potentiates release by ovine CRF, an effect blocked by the GRP receptor antagonist D-Tyr6bombesin (6-13) propylamide. Bombesin did not potentiate AVP-stimulated ACTH release; instead release was attenuated when bombesin was given at a 10-fold or greater molar excess over AVP, with no interaction seen at lower concentrations. We conclude that ovine corticotrophs express bombesin receptors, and that GRP may act in concert with other hypothalamic releasing factors to regulate ACTH secretion. PMID- 9357056 TI - Functional vasopressin V1 type receptors are present in variant as well as classical forms of small-cell carcinoma. AB - Vasopressin and other neuropeptides are believed to serve as autocrine growth factors for small-cell carcinoma of the lung (SCCL), and these mitogenic influences are reported to involve increases in intracellular Ca2+. Of the classical and variant forms of SCCL, the latter is not only more drug-resistant but also refractory to vasopressin, and other peptides, with respect to changes in intracellular Ca2+. It is currently unclear if this refractiveness of variant SCCL is due to the absence of involved peptide receptors, to the production of abnormal receptors, or to abnormalities in components of induced transduction cascades. In this study, the presence of structurally-normal and functional vasopressin V1a receptors, was examined in a classical SCCL cell line (NCI H345) that is Ca(2+)-responsive to vasopressin, and a variant SCCL cell line (NCI H82) that is unresponsive in this regard to the peptide. Both cell lines were shown to express an mRNA of 1.9 Kb for the vasopressin V1a receptor. RT-PCR, cloning, and DNA sequencing revealed the structure of the mRNA was identical for both cell lines, and, in turn, identical to the mRNA expressed for this receptor by human liver cells. In both cell lines and liver, this mRNA was shown by Western analysis and RIA to generate major protein products of approximately 70,000 and 43,000 daltons. Vasopressin action on NCI H82 cells resulted in a substantial rise in the levels of total inositol phosphates. However, it was reaffirmed that these changes in inositol phosphates were not accompanied by a rise in Ca2+ levels. All of these data indicate that variant SCCL, as well as classical SCCL, expresses structurally-normal and functional vasopressin V1a receptors, but their activation in variant SCCL raises IP3 levels without a corresponding rise in intracellular Ca2+. This difference between the two SCCL sub-types therefore involves either steps in the inositol triphosphate cascade beyond the activation of phospholipase C, or alternatively, components of other transduction events that might be involved with changes in intracellular Ca2+. PMID- 9357058 TI - Binding and biological activity of C-terminally modified melanocortin peptides: a comparison between their actions at rodent MC1 and MC3 receptors. AB - Five subtypes of melanocortin receptors have to date been identified, but to date little is known about the different structural requirements for binding and biological activity at these receptors. In this study, the role of C-terminal melanocortin peptide residues in imparting selectivity for the receptor subtypes was examined. C-terminally modified analogues of alpha-MSH and gamma-MSH were synthesized and their interaction with MC1 and MC3 melanocortin receptors was investigated. This study provides further evidence for an important role of proline 12 (numbering with respect to alpha-MSH) for binding and activity at the MC1 receptor. Although the influence of C-terminal amino acids on binding and activity at MC3-R was less marked, some of them were nevertheless observed to be beneficial for the interaction with this receptor subtype. PMID- 9357059 TI - Selectivity of cyclic [D-Nal7] and [D-Phe7] substituted MSH analogues for the melanocortin receptor subtypes. AB - The binding of the 2 cyclic lactam MSH (4-10) analogues (MTII, SHU9119), and 5 cyclic [Cys4, Cys10] alpha-MSH analogues were tested on cells transiently expressing the human MC1, MC3, MC4 and MC5 receptors. The results indicate a differential importance of the C-terminal (Lys-Pro-Val) and N-terminal (Ser-Tyr Ser) of cyclic [Cys4, Cys10] alpha-MSH analogues in binding to the MC receptor subtypes. Substitution of D-Phe7 by D-Nal(2')7 in both the cyclic lactam MSH (4 10) and the cyclic disulphide MSH (4-10) analogues resulted in a shift in favour of selectivity for the MC4 receptor; the disulphide analogue, [Cys4, D-Nal(2')7 Cys10] alpha-MSH (4-10) (HS9510), showing the highest selectivity for the MC4 receptor among all the substances tested. However, the cyclic lactams displayed an over all higher affinity for the MC receptors, than any of the cyclic disulphide MSH (4-10) analogues. PMID- 9357061 TI - Effects of aging and melatonin administration on gonadotropin-releasing hormones (GnRH) gene expression in the male and female rat. AB - It is well documented that in the rat of both sexes aging is associated with a decline in reproductive functions. We have recently shown that melatonin exerts a positive influence on GnRH gene expression in the adult male rats. In order to evaluate the effect of aging as well as melatonin on GnRH mRNA levels, we have studied the effect of 2.5-day administration of melatonin to young (50-55 day of age) and aged (18 month of age) rats of both sexes. In the young males melatonin induced a 11% increase in the hybridization signal. In the aged males, the GnRH mRNA levels were 13% lower than those observed in the young animals. Melatonin administration to aged animals completely restored GnRH mRNA levels when compared to those observed in the young untreated male rats. In contrast, melatonin did not modify the hybridization signal in young female rats, while aging induced a 20% decrease in mRNA levels. Melatonin administration to aged female induced a 18% increase in GnRH mRNA levels, thus completely reversing the influence of aging. These results indicate that the decrease in GnRH gene expression which is likely involved in the decline of reproductive functions in aging can be totally reversed by a short term administration of melatonin, then suggesting that the pineal hormone may be involved in the decrease of GnRH neuronal activity during aging. PMID- 9357060 TI - Trophic effects of melanotropin-potentiating factor (MPF) on cultures of cells of the central nervous system. AB - MPF is a tetrapeptide (structure Lys-Lys-Gly-Glu) that elicits a variety of neurotrophic effects in vivo consistent with a role in neuronal regeneration. In support of this role, we now show that MPF stimulates the proliferation of cultured astrocytes and neurite outgrowth from cultures of neocortical cholinergic and mesenchephalic dopaminergic neurons. The dose-response relationships are biphasic ("bell shaped"), maximal responses being obtained with 10(-6) M concentrations of MPF. MPF and nerve growth factor seem to act on different receptors, because their effects on cholinergic neurons are synergistic. PMID- 9357062 TI - Detection of GnRH molecular forms in brains and gonads of the crested newt, Triturus carnifex. AB - Gonadotrophin-releasing hormone (GnRH) immunoreactivity is detectable in the brain, ovary, and testis of the newt, Triturus carnifex, collected during February (reproductive phase), May, and July (nonreproductive phase). In the brain of May animals, chicken GnRH-II positive cell bodies are located within the terminal nerve, the anterior preoptic area, and the preoptic nucleus, which appears to be devoid of immunoreactive mammalian GnRH cell bodies. During February and July, both chicken GnRH-II and mammalian GnRH are detected only within the terminal nerve and anterior preoptic area. Generally, in the reproductive as well as the nonreproductive periods, chicken GnRH-II fibers are widely distributed in the brain; however, the distribution of fibers of both molecular forms suggests that they exert hypophysiotropic activity. High-pressure liquid chromatography (HPLC) coupled with radioimmunoassay indicates the presence of an early-eluting GnRH peak in brains and gonads but not in plasma. Using chicken GnRH-II antiserum, immunoreactivity is observed in spermatocytes, spermatozoa, and the external theca layer. Seasonal changes of the GnRH-like material are observed in both sexes, and its high concentration detectable during February is in good correlation with the timing of reproduction. PMID- 9357064 TI - Lactation and salt loading similarly alter neuropeptide Y, but differentially alter somatostatin, in separate sets of rat neural lobe axons. AB - Neuropeptide Y (NPY) and somatostatin immunoreactivities are present in neural lobe axons of the rat pituitary. Both peptides are upregulated during lactation, because NPY gene expression increases in the hypothalamus and plasma concentrations of somatostatin are elevated. However, the effects of lactation on NPY and somatostatin in the neural lobe are unknown. Although NPY immunoreactivity increases in the neural lobe following salt loading of male rats, the somatostatin response is unknown. To answer these questions, NPY and somatostatin immunoreactivities in the neural lobe were examined during lactation and salt loading using immunohistochemistry and image analysis. On day 2 of lactation, the area covered by immunoreactivity, a combined measurement of axon density and size of axonal swellings, of both NPY and somatostatin increased compared to ovariectomized rats. The increase in NPY was four- to fivefold greater than that of somatostatin. By day 10 of lactation, values returned to those of ovariectomized rats. Following 10 days of salt loading, the area covered by NPY immunoreactivity increased approximately 10-fold over control male rats, whereas somatostatin remained unchanged. NPY and somatostatin were not colocalized in neural lobe axons in either paradigm, demonstrating that two different neuronal populations were involved in both cases. These data indicate that NPY and somatostatin were regulated similarly during lactation, but differentially following salt loading. PMID- 9357063 TI - Effect of excitatory amino acids on rat hypothalamic somatostatin secretion in vitro. AB - We studied the effect of various agonists of excitatory amino acid (EAA) receptor subtypes on somatostatin (SRIF) release from incubated rat hypothalamic slices. N Methyl-D-aspartic acid (NMDA) and L-glutamate (1 x 10(-7) to 1 x 10(-3) M) stimulated, in a dose-dependent fashion, SRIF release. The maximal effect was obtained at a concentration of 1 x 10(-4) M for both drugs. The IC50 was 3.2 x 10(-5) M and 2.1 x 10(-5) M for NMDA and L-glutamate, respectively. Incubation with 2.5 x 10(-4) M D-2-amino-5-phosphonovalerate (a NMDA receptor antagonist) or 2-amino-4-phosphonobutyrate (a metabotropic receptor antagonist) was without significant effect on basal SRIF secretion and completely blocked the increase in SRIF release induced by 5 x 10(-5) M NMDA or L-glutamate, respectively. Incubation with 1 x 10(-4) M kainate or 0.5 x 10(-4) M alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA) did not change basal SRIF secretion. Incubation with 2 x 10(-4) M gamma-D-glutamylglycine (a specific antagonist of kainate and AMPA receptors) had no effect under basal conditions or during exposure to kainate or AMPA. Our data demonstrate that EAAs stimulate SRIF secretion in vitro, by an action through NMDA and metabotropic receptors but not kainate or AMPA receptors. PMID- 9357065 TI - Expression of adrenomedullin mRNA in the human brain and pituitary. AB - We have recently reported the presence of immunoreactive (IR) adrenomedullin (ADM) in the human brain. In the present study, the expression of ADM mRNA was studied by Northern blot analysis in the human brain and pituitary, and the presence of IR-ADM in the human pituitary was studied by radioimmunoassay. ADM mRNA was clearly detected in every region of the brain examined and in the pituitary. High concentrations of IR-ADM were present in the whole pituitary (16.7 +/- 2.0 pmol/g wet weight, mean +/- SEM, n = 4). Reverse phase high performance liquid chromatography of the pituitary showed a peak eluting in the position of human ADM(1-52). These findings suggest that ADM acts as a neuromodulator or a neurotransmitter in the brain, and as an autocrine factor, a paracrine factor, or a neurohormone in the pituitary. PMID- 9357066 TI - Gastric inhibitory polypeptide and splanchnic blood perfusion: augmentation of the islet blood flow increase in hyperglycemic rats. AB - The aim of the study was to investigate how the incretin candidate hormone gastric inhibitory polypeptide (GIP) affects splanchnic blood flow, especially pancreatic islet blood flow. For this purpose, male Sprague-Dawley rats were injected intravenously with either saline or GIP (5 or 15 micrograms/kg body weight) 10 min before blood flow measurements by a microsphere technique. Furthermore, 3 min before the blood flow measurements, 1 ml of either saline or 30% D-glucose was given intravenously. All glucose-injected animals were markedly hyperglycemic (> 20 mmol/liter) at the time of the blood flow measurements. Both doses of GIP potentiated basal and glucose-stimulated insulin release. In the normoglycemic rats, the lowest dose of GIP did not affect the blood perfusion to any of the investigated organs. The highest dose of GIP decreased whole pancreatic and duodenal blood flow, whereas islet blood flow was unaffected. As a result, fractional islet blood flow was increased. In the hyperglycemic rats, where the islet blood flow was increased compared with control animals, both doses of GIP further enhanced islet blood flow. No effect on pancreatic, fractional islet, or duodenal blood flow was seen after GIP administration to hyperglycemic animals. It is concluded that administration of GIP can further augment the glucose-induced stimulation of islet blood flow. This may contribute to facilitating release of insulin from the islets. PMID- 9357067 TI - Neurohormonal mechanisms of cigarette smoke-induced duodenal mucosal bicarbonate secretion in the rat. AB - The effect of cigarette smoke and nicotine on duodenal mucosal bicarbonate secretion (DMBS) was studied in rats. Cigarette smoke but not intravenous nicotine administered acutely to anesthetized rats via a tracheostomy tube stimulated DMBS by 47 +/- 6%. The increase was neurally mediated via atropine sensitive postganglionic cholinergic neurons. Introduction of cigarette smoke after the infusion of vasoactive intestinal peptide and porcine histidine isoleucine (PHI) also caused a delayed increase in DMBS. However, the magnitude of this increase was similar to that seen in control non-peptide-infused rats. The increase in bicarbonate secretion predominantly involved Brunner's glands. Rats exposed to cigarette smoke for 4 and 8 days before direct instillation of smoke via tracheostomy tube did not show any increase in their DMBS. These studies indicate that in the rat, cigarette smoke increases DMBS, most likely secreted by the Brunner's glands. The increase is neurally mediated via atropine sensitive postganglionic cholinergic neurons. Gastroenteric neuropeptides do not exert any influence on cigarette smoke-mediated DMBS secretion in the rat. Unlike acute exposure to cigarette smoke, chronic exposure (4 and 8 days) of rats to cigarette smoke abolishes increase in DMBS induced by subsequent exposure to cigarette smoke. This last observation may, in part, may explain the tendency of chronic smokers who have duodenal bulb ulcers to show greater propensity to higher rate of recurrence and protracted healing. PMID- 9357068 TI - Gastrin-releasing peptide stimulates possum gallbladder contractility in vitro. AB - Reports of the action of gastrin-releasing peptide (GRP) on gallbladder contraction are limited to a few species. We compared the action of GRP, acetyl gastrin-releasing peptide 20-27 (GRP 20-27), and cholecystokinin-octapeptide (CCK 8) on gallbladder contractility with and without pretreatment with the neural inhibitor tetrodotoxin (TTX) and the bombesin antagonist [D-Phe 6, Des-Met 14] bombesin 6-14 ethylamide (BBS 6-14 E). Full-thickness muscle strips were prepared and suspended in organ baths. The maximum GRP, GRP 20-27, and CCK-8 responses were 54.2 +/- 4.2%, 74.6 +/- 6.4%, and 69.3 +/- 6.9% of that of carbachol, respectively. Pretreatment with TTX influenced the action of GRP 20-27, and pretreatment with BBS 6-14 E influenced that of GRP and GRP 20-27. These studies show that GRP and GRP 20-27 are potent agonists of gallbladder contractility, acting via GRP-preferring receptors, and that GRP 20-27 also acts via a neural component. PMID- 9357069 TI - Processing of [D-Arg1,D-Phe5,D-Trp7,9,Leu11]substance P in xenograft bearing Nu/Nu mice. AB - [D-Arg1,D-Phe5,D-Trp7,9,Leu11]Substance P is a broad-spectrum neuropeptide growth factor antagonist that has exhibited in vitro activity against a range of human cancer cell lines. The fate of this compound in vivo following i.p. administration at 12 micrograms/g to nu/nu mice bearing the H69 small-cell lung cancer xenograft has been studied. Metabolism was confined to the C-terminus producing [D-Arg1,D-Phe5,D-Trp7,9,Leu11]substance P acid and [D-Arg1,D-Phe5,D Trp7,9]substance P(1-10). The peptide had a long half-life in plasma (45.9 min) and became widely distributed among the tissues studied with the highest accumulation observed in the liver (AUC 1102 micrograms/g x min) and the lowest in the brain (5 micrograms/g x min). Uptake into the tumor xenograft was poor (AUC 189 micrograms/g x min); however, uptake into the lungs was much greater (AUC 507 micrograms/g x min), offering encouragement that therapeutic concentrations may be targeted to primary lung tumors. PMID- 9357070 TI - Role of adrenomedullin and related peptides in the regulation of the hypothalamo pituitary-adrenal axis. AB - Adrenomedullin (ADM) is a hypotensive peptide, originally isolated from human pheochromocytomas, and then found to be widely distributed in the various body systems. ADM derives from preproadrenomedullin, a 185-amino acid residue prohormone, containing at its N-terminal a 20-amino acid sequence, named proadrenomedullin N-terminal 20 peptide (PAMP). ADM and PAMP immunoreactivities have been detected in the hypothalamo-pituitary-adrenal (HPA) axis of humans, rats, and pigs. Adrenal glands possess binding sites for both ADM and PAMP, the former being mainly of the subtype 1 of calcitonin gene-related peptide (CGRP) receptors. ADM exerts a direct inhibitory action on angiotensin II- or potassium stimulated aldosterone secretion of zona glomerulosa cells. This effect is mediated by the CGRP1 receptor and its mechanism probably involves the blockade of Ca2+ influx. In contrast, ADM enhances aldosterone production by in situ perfused rat adrenals and human adrenal slices (containing medullary chromaffin cells), again through the activation of CGRP1 receptors. This aldosterone secretagogue effect of ADM is blocked by the beta-adrenoceptor antagonist l alprenolol, thereby suggesting that it is indirectly mediated by the release of catecholamines by chromaffin cells. The effects of ADM on adrenal glucocorticoid release are doubtful and probably mediated by the increase in adrenal blood flow rate and the inhibition of ACTH release by pituitary corticotropes. The concentrations reached by ADM and PAMP in the blood rule out the possibility that they act on the HPA axis as circulating hormones. Conversely, their content in both adrenal and hypothalamo-pituitary complex is consistent with a paracrine mechanism of action, which may play a potentially important role in the regulation of fluid and electrolyte homeostasis. PMID- 9357071 TI - Experimental diabetes upregulates the expression of uretereral endothelin receptors. AB - We investigated the binding characteristics of endothelin (ET) receptors in the ureters of rats with experimentally induced diabetes and diuresis. Receptor binding experiments demonstrated an upregulation in the expression of [125I]ET-1 binding sites in the diabetic rat ureter but not in the diuretic rat ureter. ET 1, ET-3, IRL 1620, and BQ 610 inhibited [125I]ET-binding to the rat ureter consistent with the predominance of ETA receptors in these tissues. The subtype specificity of ET receptors in ureteral tissues was confirmed with inhibition data obtained from cloned human ETA and ETB receptors. PMID- 9357073 TI - People who move: new reproductive health focus. PMID- 9357072 TI - Melanin-concentrating hormone (MCH) involvement in pentylenetetrazole (PTZ) induced seizure in rat and guinea pig. AB - Intraperitoneal injection of 60 mg/kg of pentylenetetrazole (PTZ) induced seizure in rats, but was subthreshold and did not result in seizure in guinea pigs. Three days after intracerebroventricular (i.c.v.) injection of 75 micrograms 6 hydroxydopamine (6-OHDA) in guinea pigs, PTZ induced seizures similar to those seen in rats. In both rats and 6-OHDA-treated guinea pigs, i.c.v. injection of melanin-concentrating hormone (MCH) 15 min before PTZ prevented seizure activity. These results suggest that MCH-containing neurons may participate in the neural circuits involved in expression of PTZ-induced seizure. PMID- 9357074 TI - Improvement of a respiratory ozone analyzer. AB - The breath-to-breath measurement of total respiratory ozone (O3) uptake requires monitoring O3 concentration at the airway opening with an instrument that responds rapidly relative to the breathing frequency. Our original chemiluminescent analyzer, using 2-methyl-2-butene as the reactant gas, had a 10% to 90% step-response time of 110 msec and a minimal detectable concentration of 0.018 parts per million (ppm) O3 (Ben-Jebria et al. 1990). This instrument was suitable for respiratory O3 monitoring during quiet breathing and light exercise. For this study, we constructed a more self-contained analyzer with a faster response time using ethylene as the reactant gas. When the analyzer was operated at a reaction chamber pressure of 350 torr, an ethylene-to-sample flow ratio of 4:1, and a sampling flow of 0.6 liters per minute (Lpm), it had a 10% to 90% step response time of 70 msec and a minimal detectable concentration of 0.006 ppm. These specifications make respiratory O3 monitoring possible during moderate-to heavy exercise. In addition, the nonlinear calibration and the carbon dioxide (CO2) interference exhibited by the original analyzer were eliminated. In breath to-breath measurements in two healthy men, the fractional uptake of O3 during one minute of quiet breathing was comparable to the results obtained by using a slowly responding commercial analyzer with a quasi-steady material balance method (Wiester et al. 1996). In fact, fractional uptake was about 0.8 regardless of O3 exposure concentration (0.11 to 0.43 ppm) or ventilation rate (4 to 41 Lpm/m2). PMID- 9357075 TI - Record high U.S. life expectancy. AB - In 1996 life expectancy in the United States rose to a record high of 75.9 years for all persons combined. A new peak was also recorded among newborn boys--72.8 years--while average future lifetime for infant girls increased to 79.0 years- 0.1 year shy of the 1992 record of 79.1 years. In recent years longevity gains among males have outpaced those for females, with the result that the sex differential in longevity at birth in favor of females has narrowed considerably. In 1996 newborn girls could anticipate living 6.2 years longer than boys--the gap was 6.8 years in 1992 and 7.0 years in 1990. Projections indicate that the trend in longevity improvements in favor of males will continue. As a consequence, the sex differential gap in average future lifetime is anticipated to diminish to 4.6 years by 2050. PMID- 9357076 TI - Variations in average charges for strokes and TIAs: United States, 1995. AB - The 1995 in-hospital charges for 6,628 group health insured stroke victims averaged $11,010 across the country. This total was over twice the average charge for the 1,584 patients hospitalized with a transient ischemic attack (TIA), $4,940. The Mountain and the neighboring Pacific areas of the country reported the highest charges for a stroke, 24 and 16 percent, respectively, higher than the U.S. average. The charges in the East North Central and East South Central areas were the lowest, each under $9,000 and 20 and 25 percent below the norm, respectively. Between study states, the highest stroke charge was reported in Arizona ($17,590) and the lowest in Ohio ($6,670). Hospital charges comprised 81 percent of the total bill to insurance, averaging $8,940. Physicians' charges averaged $2,070, with those in New York 34 percent above the norm and those in Alabama 30 percent below ($1,450). The New Jersey hospital stays averaged 8.1 days, whereas the stay in Oregon was 5.2 days. The total TIA charge was just under $5,000 across the country. Illinois reported the highest TIA in-hospital charge, $6,160, 25 percent above the U.S. average and almost twice the total in Alabama ($3,170). The hospital charges comprised 87 percent of the total, averaging $4,290. Physicians' charges in Pennsylvania were the highest ($890, 37 percent above the U.S. norm of $650) and those in Alabama the lowest ($450, 31 percent below). The average length of stay was 3.7 days for a TIA, ranging from 5.4 days in New York to 2.3 days in Arizona. PMID- 9357077 TI - Trends in stimulation and induction of labor 1989-1995. AB - Between 1989 and 1995 the rates for stimulation and induction of labor rose every year, representing a 48 and 77 percent, respectively, total rise over the time period. In 1995 the rate of stimulation was 161 per 1,000 live births and of induction 160. Two percent, or 74,167, of the 3,899,589 births in 1995 had both procedures performed. While rates of stimulation decline with advancing maternal age, the induction rates tend to be higher for older women. Rates for both procedures increased between 1989 and 1995 for both black and white women in all age categories. Women whose pregnancies have extended beyond the expected gestation of 37 weeks consistently had much higher rates of both stimulation and induction. Rates for both procedures rose for doctors of medicine (MD's), doctors of osteopathy (DO's) and certified nurse-midwives (CNM's). DO's had the greatest increases in both stimulation and induction rates. Declines in the cesarean section rate were greater for births that were stimulated or induced than for those without either of these procedures. The rates for stimulated or induced vaginal birth after cesarean (VBAC) were double those of VBACs without such procedures. PMID- 9357078 TI - Personal income and finances: regional comparisons. AB - As the "baby boom" generation moves closer to retirement and Social Security and Medicare face potential problems, the financial condition of U.S. households and families has become a much discussed topic. The personal saving habits, wealth and debt burden of society have been intensely analyzed in order to gauge the future impacts on the economy. This article views personal income growth from a regional perspective, pointing to the vastly differing outcomes of total and per capita data. The importance of regional differences in cost of living and taxation and the possible impacts on personal finances are highlighted. A review and debate on the broad brush approach of many federal programs is suggested. PMID- 9357079 TI - A century of suicide in Italy: a comparison between the old and the young. AB - This study analyzes suicide rates from 1887 to 1993 in the Italian population between the ages of 15 and 24 years old and over 65 years of age, based on official data published in the Health Statistics Year Book. The rates of death by suicide (per 100,000) subjects) are calculated for each year and for 10-year periods, as are the mortality rates relative to each method of suicide, standardized by gender. The latter analysis was possible starting from 1951 only, when it became customary to record method. The findings indicate an increase in the suicide phenomenon in the elderly population in Italy over the test period. Rates are at least 3 times higher for men than for women. The highest rates are reported for elderly men, but there appears to have been a greater proportional increase in the number of suicides committed by elderly women. The rise was statistically significant in both males and females. By contrast, a rather constant decrease in suicide rates in young people emerges from the beginning of the century through to the present date. This decrease is more marked in females, although suicide rates are lower for females than for males. Over the study period, substantial changes have come about in the suicide methods used by both young and old people. There was an increase in poisoning by care exhaust fumes, jumping from heights, hanging, and firearms. PMID- 9357081 TI - Temporal trends and geographic patterns of teen suicide in Alaska, 1979-1993. AB - The author analyzed death certificate and U.S. census data to document trends in suicide rates among Alaskans 14-19 years of age. During 1979-1993, Alaskan teenagers had a suicide rate of 31.5 per 100,000 persons per year. Suicide rates varied up to sixfold by race, gender, and local census area of residence; in particular, Alaska Native males had one of the highest documented suicide rates in the world. Suicide rates increased two- to threefold during the study period for persons less than 18 years of age, while remaining stable for older teenagers. Within census areas, suicide rates correlated inversely with the percentage of all households headed by a married couple. PMID- 9357080 TI - Youth suicide in Norway, 1990-1992: a comparison between children and adolescents completing suicide and age- and gender-matched controls. AB - In Norway 1990-1992, the suicide rate was 18.6 per 100,000 individuals per year for boys 15-19 years old and 6.3 for girls, and for 10-14 year olds the rate was 2.7 for boys and 0.5 for girls. Comparison of all completed suicides (N = 129) with gender- and age-matched control subjects identified depression (OR = 19.9; CI = 11.2, 35.5), disruptive disorders (OR = 6.0; CI = 3.1, 11.4), and previous suicidal behavior (OR = 3.4, CI = 2.0, 5.6) as main risk factors. Of the suicide completers, 74% had mental disorders. Suicidal intent was previously expressed by 48%, but few (24%) had received treatment, despite well-developed health services. A history of disruptive disorders (17%) and substance abuse (10%) were less frequently found than in previous studies, but binge drinking may contribute to the adolescent suicide rate. PMID- 9357082 TI - Adolescents' misperceptions of the dangerousness of acetaminophen in overdose. AB - Clinical observations suggest that adolescents commonly and naively use acetaminophen in suicide attempts even when they do not wish to die. It is estimated that 18,500-mg acetaminophen tablets can lead to hepatotoxicity, while death is usually associated with ingestion of 50 or more tablets. A sample comprising 569 adolescent students completed an author-designed survey assessing teenagers' knowledge of acetaminophen's therapeutic and harmful effects. The findings support our original data that adolescents have ready access to acetaminophen and use it in suicide attempts, but underestimate its potential for toxicity. Forty-two percent of this sample underestimated the dose to cause harm, believing it would require 20 or more tablets, and 50% underestimated the dose to cause death, stating 100 or more pills would be necessary. Adolescents appear to seriously underestimate the dangerousness of acetaminophen in overdose, and lack knowledge regarding side effects of overdose. PMID- 9357083 TI - Adolescent Multiphasic Personality Inventory and its utility in assessing suicidal and violent adolescents. AB - Adolescent Multiphasic Personality Inventory profiles of inpatient adolescents were examined to identify differences between suicidal (danger to self, n = 145) and violent (danger to others, n = 36) adolescents. Participants were 181 inpatients (12-17 years old, mean age, 14.7 years) admitted to an adolescent psychiatric unit of a southern California county hospital. Results indicate that the suicide group had significantly higher scores than the violent group on five scales (Hypochondriasis, Psychasthenia, Paranoia, Schizophrenia, and Social Introversion). This scales, singly or in combination, have been found to be indicative of psychotic process. This suggests the presence of psychotic process in suicidal, but not violent, adolescents. PMID- 9357084 TI - The Big Ten Student Suicide Study: a 10-year study of suicides on midwestern university campuses. AB - The Big Ten Student Suicide Study was undertaken from 1980-1990 to determine the suicide rates on Big Ten University campuses. The study design attempted to address many of the statistical and epidemiological flaws identified in previous studies of campus student suicides. The 10-year study collected demographic and correlational data on 261 suicides of registered students at 12 midwestern campuses. The largest number of suicides for both males and females were in the 20-24-year-old age group (46%), and amongst graduate students (32%). The overall student suicide rate of 7.5/100,000 is one half of the computed national suicide rate (15.0/100,000) for a matched sample by age, gender, and race. Despite the overall lower suicide rate, the analyses revealed that students 25 and over have a significantly higher risk than younger students. Although women have rates roughly half those of men throughout their undergraduate years, graduate women have rates not significantly different from their male counterparts (graduate women 9.1/100,000 and graduate men 11.6/100,000). PMID- 9357085 TI - The effectiveness of suicide prevention centers: a review. AB - This article reviews 14 studies examining whether suicide prevention centers have a preventive effect on suicide rates. Seven studies were identified that provide some support for a preventive effect, one found an increase in the suicide rates, and six failed to find any significant effects (either preventive or facilitative). The studies' different methodologies are reviewed, and limitations on the authors' conclusions pointed out. The conclusion of this article is that the evidence provides support for a preventive effect from suicide prevention centers, albeit small and inconsistently found. PMID- 9357086 TI - John Holland: the near-death of a frightened sailor: specific phobia and suicide related behavior. PMID- 9357088 TI - What's in a name... PMID- 9357087 TI - Survivor siblings, the case of schizophrenia: a response to "Impact of adolescent suicide on siblings and parents" by Brent, Moritz, Bridge, Perper, and Canobbio. PMID- 9357089 TI - Vigabatrin serum concentration to dosage ratio: influence of age and associated antiepileptic drugs. AB - The relationship between the ratio of vigabatrin concentration to dosage (VGB C/D) and both patient age and the presence of other antiepileptic drugs (AEDs) was analyzed retrospectively by bivariate and multivariate methods in 179 patients with epilepsy (114 children and 65 adults). Of the 179 patients, 33 received VGB alone (30 children and 3 adults) and 146 received VGB with other AEDs (84 children and 62 adults). Vigabatrin trough steady-state serum concentration correlated better with VGB dosage in milligrams per kilogram than the dosage in milligrams in children (r = 0.62 vs. r = 0.17, P < 0.001) but not in adults (r = 0.51 vs. r = 0.49, NS). The correlation between milligrams per kilogram and serum concentration of VGB was greater in children on monotherapy (r = 0.83) than in those on polytherapy (r = 0.46). Vigabatrin C/D ratio increased significantly with age (r = 0.51, P < 0.001), being lower in children than in adults both by Student's t-test (0.087 +/- 0.039 vs. 0.128 +/- 0.057, mean +/- SD, P < 0.001) and by two-way analysis of variance when controlling for other AEDs (P < 0.001). Inducing AEDs seemed to increase VGB C/D ratio in the bivariate tests, but this influence decreased and even disappeared if patient age was considered in the multivariate analysis. However, the increase in VGB C/D ratio with VPA serum concentration (r = 0.46, P < 0.001) was confirmed by multiple regression including age (P < 0.001). Intrapatient variability of VGB C/D ratio was 29 +/- 18%. It was concluded that trough steady state VGB serum concentration may be more predictable in children based on the milligrams per kilogram dosage than on the milligram dosage, and that the influence of patient age should be considered if the VGB C/D ratio is used to estimate patient compliance. PMID- 9357091 TI - Patterns of azathioprine metabolites in neutrophils, lymphocytes, reticulocytes, and erythrocytes: relevance to toxicity and monitoring in recipients of renal allografts. AB - Monitoring of azathioprine (AZA) therapy by the measurement of 6-thioguanine nucleotides (6-TGN) concentrations in red blood cells (RBC) may improve safety and ensure optimal immunosuppressive effects of AZA in organ transplantation. The authors explored the rationale for such monitoring by measuring thiopurine metabolites in peripheral blood cell types that are more relevant to the effects and kinetics of AZA and its active metabolites. Neutrophil granulocytes were isolated by density gradient centrifugation, and CD4+ lymphocytes and reticulocytes by using specific immunomagnetic beads. In neutrophils, 6-TGN concentrations had median measurements 31 times higher than in RBCs. In contrast to the high methylated mercaptopurine (me-MP) concentrations in RBCs, these metabolites were not detected in the neutrophils. Thiopurine metabolite levels were lower than the analytic limit of detection in all the CD4+ samples. The concentrations of 6-TGN and me-MPs were lower in reticulocytes than in RBCs in general, indicating that thiopurine metabolites are taken up by RBCs in the circulation. This study's findings, that 6-TGN concentrations are very high in neutrophils, whereas me-MPs are undetectable, many explain the specific neutropenic adverse effect of AZA. The results also add support to monitoring AZA through measurements of 6-TGN and me-MPs in RBCs. PMID- 9357090 TI - No increase in carbamazepine-10,11-epoxide during addition of lamotrigine treatment in children. AB - It has been suggested that lamotrigine (LTG) may enhance the toxicity of carbamazepine (CBZ) by increasing the concentration of the active metabolite carbamazepine-10,11-epoxide (CBZ-E) in adult patients. The authors investigated this hypothesis in an add-on study in 11 children and 3 adolescents, aged 6-22 years, who had been treated for more than 1 year with CBZ in monotherapy or with CBZ in combination with one or two other antiepileptic drugs. The LTG dosage was increased step by step until clinical response or side effects were observed. The plasma concentrations of LTG, CBZ, and CBZ-E were monitored during steady state conditions before and after the addition of LTG. It was found that LTG had no effect on mean CBZ concentrations and that it decreased rather than increased the mean plasma concentration of CBZ-E from 6.4 +/- 2.6 to 4.9 +/- 2.4 mumol/l (mean +/- SD, n = 14, P = 0.019). Observed side effects were diplopia in two children, agitation in two, and increased number of seizures in one. None of these five patients had unusually high CBZ-E levels when the side effect developed. It is concluded that addition of lamotrigine in children treated with carbamazepine children does not result in a pharmacokinetic interaction with a toxic accumulation of carbamazepine-10,11-epoxide. PMID- 9357092 TI - Digoxin intoxication in a patient with end-stage renal disease: efficacy of digoxin-specific Fab antibody fragments and peritoneal dialysis. AB - Digoxin intoxication is a serious medical problem, and impairment of renal function is a common risk factor for toxicity. Digoxin specific antibody fragments (Fab) is the most effective treatment available for severe digitalis intoxication. The use of Fab therapy in a patient with renal disease is considered as effective as in patients with normal renal function, although the increased risk of rebound digoxin toxicity mandates a longer period of observation. In patients with kidney failure, neither digoxin nor Fab can be removed efficiently from the systemic circulation by hemodialysis or continuous arteriovenous hemofiltration. Knowledge about the clearance of both compounds by peritoneal dialysis is limited. The authors describe a patient with end stage renal disease who was treated with Fab and peritoneal dialysis for life threatening digoxin intoxication. Like other forms of dialysis, peritoneal dialysis, even when performed in an intensive schedule, is not associated with an enhanced clearance of digoxin. PMID- 9357094 TI - Caffeine in saliva after peroral intake: early sample collection as a possible source of error. AB - The influence of collection time on the correlation of caffeine concentrations in saliva and serum was examined in six healthy adults after peroral administration of 5 mg/kg caffeine citrate. Saliva was obtained from three different salivary glands (sublingual, right parotid, and left parotid) and evaluated separately. Caffeine concentrations in saliva and serum samples were determined by high performance liquid chromatography. There were no differences in the caffeine concentrations in saliva from the three investigated glands (alpha = 0.05). Saliva samples collected earlier than 2 hours after caffeine intake showed higher caffeine concentrations than could be expected from the corresponding serum samples. Gingiva contamination was shown to be responsible for the higher caffeine concentrations in saliva, and it was concluded that saliva is a feasible matrix for therapeutic drug monitoring of caffeine. If caffeine is administered orally, saliva samples should be taken at least 2 hours after caffeine intake. If caffeine-containing beverages are used as the source of caffeine or if subjects do not cooperate by rinsing the mouth of caffeine contamination, an additional 60 minutes should be added before saliva sampling. PMID- 9357093 TI - Lack of kinetic interaction between valproic acid and citrus pectin. AB - To determine the influence of ingestion of dietary fiber (citrus pectin) in the kinetic disposition of valproic acid, two studies were conducted in adult volunteers. Twelve healthy subjects took single oral doses of 500 mg valproic acid in a form of sodium valproate as coated tablets, on two occasions: under fasting conditions (treatment A) or immediately after ingestion of 14 g citrus pectin (treatment B). In each study, a randomized, single-dose, two-way crossover design was used, and 8 days elapsed between the two trials. Assays were carried out for valproic acid in plasma samples, which were collected from zero to 48 hours after drug administration. The peak plasma concentration (60.52 vs. 55.74 micrograms x ml-1), the time to peak concentration (3.47 vs. 3.91 h), the absorption rate constant (0.12 vs. 0.12 h-1), and the area under the plasma concentration-time curve (1324.0 vs. 1268.5 micrograms x h x ml-1) did not differ significantly between treatments A and B (median, Wilcoxon test). These results suggest that the administration of citrus pectin does not alter the rate and extent of absorption of valproic acid administered as a single dose to healthy volunteers. PMID- 9357096 TI - Correlation of morphine sulfate in blood plasma and saliva in pediatric patients. AB - This study sought to determine whether saliva concentrations of morphine correlate with plasma levels of morphine in pediatric patients receiving morphine analgesia for severe pain, and to evaluate whether the measurement of saliva morphine concentrations would be a useful, noninvasive, clinical tool to diagnose systemic exposure to morphine. Fifteen pediatric patients were enrolled; for the control group, 18 adult volunteers were recruited. Patients received continuous morphine drips to ameliorate pain caused by a sickle cell vasoocclusive crisis (range, 10-40 micrograms/kg.h). Control subjects were randomized into those receiving acetaminophen with either 8 mg (n = 13) or 30 mg (n = 5) of codeine. All participants fasted at least 2 hours before sample collection. Blood and saliva samples were collected simultaneously. All samples were analyzed by radioimmunoassay for morphine. There was no correlation between saliva and plasma morphine concentrations in either the patients receiving intravenous morphine (r = 0.04, P = 0.89) or in the controls receiving codeine (r = 0.43, P = 0.08). There was no observed difference in the mean counts per minute (CPM) for saliva samples in the pH range 3.96 to 8.06. Saliva concentrations of morphine cannot be used to predict the plasma concentration of morphine in children or adults. However, the concentration of morphine in saliva may be used as a qualitative indicator of systemic exposure to morphine in a subject. PMID- 9357095 TI - Plasma itraconazole concentrations in patients with neutropenia: advantages of a divided daily dosage regimen. AB - A group of 36 patients in the hematology department of Saint-Antoine Hospital, Paris, France, was on chemotherapy. The patients were also given antiacid drugs to prevent gastrointestinal toxicity and itraconazole as prophylaxis against aspergillosis. The antifungal drug was given as 100-mg capsules three times a day shortly after meals. The plasma itraconazole and hydroxyitraconazole concentrations were measured by high-performance liquid chromatography at steady state. Of 36 patients, 29 (81%) had adequate plasma itraconazole concentrations (> or = 250 ng/ml) after 8 +/- 2 days. The 7 patients with low plasma itraconazole concentrations were given 200 mg three times a day. Of the 36 patients, 34 (94%) had effective plasma concentrations within 2 weeks of the beginning of treatment. The two remaining patients were lost to follow-up. The proposed itraconazole regimen provides effective prophylaxis against aspergillosis and represents a substantial economic advantage over a single daily dose of 400 to 600 mg. The marked intrapatient and interpatient variations in plasma itraconazole indicate the need for regular therapeutic drug monitoring to ensure effective plasma itraconazole concentrations in all neutropenic patients. PMID- 9357097 TI - Elevation of plasma carbamazepine concentrations by ketoconazole in patients with epilepsy. AB - The effect of ketoconazole (200 mg/d orally for 10 days) on the plasma concentrations of carbamazepine (CBZ) and its active metabolite carbamazepine 10,11-epoxide (CBZ-E) was assessed in eight patients with epilepsy stabilized on CBZ therapy. Administration of ketoconazole was associated with a significant increase in plasma CBZ concentrations (from 5.6 +/- 1.9 to 7.2 +/- 2.9 micrograms/ml on day 10 [means +/- SD, P < 0.02]), whereas plasma concentrations of CBZ-E were unchanged. After ketoconazole was discontinued, plasma CBZ levels decreased to pretreatment values. This interaction was probably mediated by an inhibiting action of ketoconazole on cytochrome CYP3A4, the main enzyme responsible for CBZ metabolism. PMID- 9357098 TI - Quantification of the O- and N-demethylated and the glucuronidated metabolites of codeine relative to the debrisoquine metabolic ratio in urine in ultrarapid, rapid, and poor debrisoquine hydroxylators. AB - The O-demethylation of codeine is polymorphic and catalyzed by CYP2D6. The metabolites of codeine formed through O- and N-demethylation as well as glucuronidation were quantified in the ultrarapid metabolizers of debrisoquine and compared with the normal extensive (EM) and poor metabolizers (PM). The urinary codeine and its seven metabolites were detected after 25 mg codeine in 24 healthy Caucasian subjects with low debrisoquine metabolic ratios (MR, < or = 0.11) and a group of 132 subjects tested earlier with codeine and debrisoquine including 114 EMs (MR < 12.6) and 18 PMs (MR > 12.6). Whereas the O-demethylated metabolites accounted for < 0.4% of the total recovery on average in the PMs and 1.7% to 8.7% in the EMs, they accounted for 15.3% in the 24 subjects with ultrarapid metabolism of debrisoquine. This study suggests that the ultrarapid debrisoquine hydroxylators may develop increased O-demethylated metabolite dependent effects or side-effects of codeine. PMID- 9357099 TI - Quick onset of severe abdominal pain after codeine in an ultrarapid metabolizer of debrisoquine. AB - The authors describe a 33-year-old woman who experienced severe pain in the epigastrium after codeine intake. This side-effect is consistent with that of morphine. Later, the patient was phenotyped and genotyped as an ultrarapid metabolizer with high capacity to metabolize codeine to morphine. PMID- 9357100 TI - Measurement of low serum methotrexate concentrations with the Syva Emit Methotrexate Assay. PMID- 9357101 TI - Antisense oligonucleotides: a new therapeutic approach. AB - The use of antisense oligonucleotides as therapeutic agents has heralded a new field of genetic pharmacology. Oligonucleotides are relatively easy to design and display increased affinity and selectivity for their nucleic acid targets compared with traditional drugs. However, the development of antisense therapy has not been as simple as was first believed; many critical issues had to be addressed. The first generation of oligonucleotides investigated as drug candidates were phosphorothioate oligonucleotides. Several animal experiments have provided evidence that antisense oligonucleotides can inhibit gene expression of disease-associated proteins. These promising studies led to the advancement of these compounds into clinical trials in such diverse fields as infectious diseases, cancer and inflammation. The insights gained through ongoing clinical trials has opened the pathway to the design of second-generation oligonucleotides which have an improved safety profile and efficacy. PMID- 9357102 TI - Timed ELISA: an alternative approach to quantitative enzyme-linked immunosorbent assay. AB - As the uses for ELISA (enzyme-linked immunosorbent assay) increase, so does the need for a quantitative procedure that does not require a spectrophotometer or other expensive equipment. 'Timed ELISA' employs an 'iodine clock' as the final step such that quantitative measurements may be made using a stopwatch. Catalase, coupled to the primary antibody, reduces the concentration of H2O2 available to generate iodine in the clock reaction. Iodine stains the starch component blue, but catalase prolongs the time taken for the change in colour to be observed. After the time delay occurs the transition to full colour development is extremely rapid (< 1 s) at all analyte concentrations, allowing clear definition of the end point. The performance of Timed ELISA is similar to that obtained using a horseradish peroxidase-conjugated system employing the customary spectrophotometric determination. PMID- 9357103 TI - Characterization of the acidic oligosaccharides assembled on the Pichia pastoris expressed recombinant kringle 2 domain of human tissue-type plasminogen activator. AB - The N-linked glycans assembled in Pichia pastoris on the recombinant kringle 2 domain of human tissue-type plasminogen activator (r-[K2tPA]) are composed of approx. 80% neutral and 20% charged species. After peptide:N4-(N-acetyl-beta glucosaminyl)asparaginyl amidase-catalysed liberation of the oligosaccharides from the purified glycopeptide, the glycan mixture was resolved by HPLC on amino silica-based resin. Oligosaccharide mapping of the resulting mixture by HPLC, gel filtration and time-of-flight matrix-assisted laser-desorption-ionization-with delayed-extraction mass spectrometry (TOF-MALDI DE-MS) revealed that > 90% of the charged species consisted of a series of oligosaccharides possessing molecular masses that were consistent with a range of saccharides comprising phospho Man10GlcNAc2-phospho-Man14GlcNAc2, with phospho-Man11GlcNAc2 representing the major species. The remaining material in the charged fraction contained identifiable phosphorylated glycans that were one or two mannose units shorter, and one to four mannose units longer, than those present in the above range of oligosaccharides. Treatment of the native charged glycan pool with alkaline phosphatase did not result in molecular-size alterations, showing that phosphomonoesters are not present. Mild acid hydrolysis of the glycans led to a decrease in the size of all charged glycans by one mannose residue, providing phospho-Man9GlcNAc2-phospho-Man13GlcNAc2. Following this procedure, treatment with alkaline phosphatase resulted in size decreases that were equivalent to the loss of one phosphate group from each glycan. This demonstrates that all charged glycans isolated contained phosphate in phosphodiester bonds to two mannose units. The present study shows that P. pastoris cells possess the capability of assembling phosphorylated glycans having the phosphate moiety present in phosphodiester linkages with two mannose units. These saccharides, like the neutral oligosaccharides, contain considerably smaller amounts of mannose than glycans present in other strains of yeast. PMID- 9357104 TI - Application of a statistical technique to the production of Saccharomyces cerevisiae (baker's yeast). AB - The identification of a static model for Saccharomyces cerevisiae (baker's yeast) production under aerobic conditions has been realized. Two-level factorial experimental design was used to identify a statistical model. In order to find the optimal operating conditions by using the statistical model, Box-Wilson's steepest-ascent method was used [Box and Wilson (1951) J. R. Stat. Soc. Ser. B 13, 1]. Four independent variables which have an effect on aerobic yeast production were selected. These were temperature, pH, air flow rate and agitation rate. Yeast productivity was selected as the dependent variable. A first-order statistical model was considered to illustrate the dependence of the yeast productivity on the operating parameters. Optimum pH, temperature, air flow rate and agitation rate were determined as 5, 32 degrees C, 1 vol./vol. per min and 600 rev./min respectively. PMID- 9357105 TI - Enhanced production of cellobiose dehydrogenase in cultures of Phanerochaete chrysosporium supplemented with bovine calf serum. AB - The enzyme cellobiose dehydrogenase (CDH), produced by many wood-degrading fungi has, in recent years, attracted considerable interest for its possible role in both cellulose and lignin degradation. To characterize the enzyme better and to identify its role in the degradation of wood and wood components, it is desirable to produce it in higher amounts. We report here that the addition of bovine calf serum to cellulose-grown cultures of Phanerochaete chrysosporium enhances the production of certain enzymes, CDH in particular. The highest CDH production was obtained with 45 ml of serum/litre of medium added on day 3 or 4. The resultant CDH yield was approx. 700-800 units/litre, which was 3.5-4 times higher than that in cultures without serum. Serum addition also enhanced the production of beta glucosidase. However, the impact on CDH production was the most dramatic. The enhanced enzyme production cannot be explained by increased rates of spore germination, simple nutrient effects or cofactor effects. Fractionation of serum by Cohn's fractionation technique showed that the albumin (BSA) fraction had almost the same effect as whole serum. However, purified BSA had less effect than crude BSA (fraction V of Cohn's fractions), suggesting that an additional factor, probably a protease inhibitor in serum, also contributed to the effect of serum. PMID- 9357106 TI - Conjugation of catalase to a carrier antibody via a streptavidin-biotin cross linker. AB - Targeting of catalase could be useful in antioxidative protection of cells challenged with H2O2. In the present study we conjugated catalase to a carrier model antibody using a biotin-streptavidin (SA) cross-linker and characterized the functional activity of the conjugate. Neither biotinylation nor conjugation with SA decreased the enzymic activity of catalase. Further coupling of radiolabelled biotinylated catalase (b-catalase) to biotinylated antibody (b-Ab) via SA using a two-step conjugation procedure did not change enzymic activity of b-catalase. b-Ab-SA-b-catalase specifically bound to antigen-coated plastic wells, but not to albumin-coated plastic wells. Substitution of b-Ab with control biotinylated IgG (b-IgG) abrogated binding of the catalase to the antigen. H2O2 was degraded in antigen-coated wells preincubated with b-Ab-SA-b-catalase, but not with b-IgG-SA-b-catalase. Thus b-Ab-SA-b-catalase specifically binds to immobilized antigen and degrades H2O2 after binding to the target. The methodology described in the present paper may be useful for the development of a novel strategy for antioxidant therapy. PMID- 9357108 TI - Purification of mouse H1 histones expressed in Escherichia coli. AB - We amplified the coding regions of the previously cloned H1 genes H1-1, H1 zero and H1t and inserted them into the expression vector pET-11d. The synthesis of the H1 histones can be induced in the appropriate strains of bacteria, and the H1 histones can be readily purified. We report detailed protocols for the purification of the expressed proteins using combinations of ion-exchange and reverse-phase HPLC. Sufficient amounts of each pure variant protein can be obtained for use in physical studies of H1-DNA interactions. PMID- 9357107 TI - Purification and characterization of lipase from a raw-milk yeast (Trichosporon asteroides). AB - A lipase-producing yeast strain was isolated from raw milk. It was identified as Trichosporon asteroides strain LP005. The lipase from this yeast was purified 8 fold to homogeneity for further characterization. The purification process for this lipase included (NH4)2SO4 precipitation at 70% saturation and gel filtration on Sephadex G-200. The molecular mass of the lipase was 37 kDa as determined by SDS/PAGE. The optimum pH and temperature for activity were pH 5.0 and 60 degrees C. The lipase was stable over a wide pH range (3.0-10.0) and at temperatures lower than 70 degrees C. The chelating agent EDTA did not affect activity of the enzyme, and this suggested that it was not a metalloenzyme. Treatment of tuna oil with T. asteroides LP005 lipase gave approx. 35 and 47% increases in the concentration of eicosapentaenoic acid and docosahexaenoic acid respectively. Thus, this lipase could potentially be used for the concentration step in the production process of such polyunsaturated fatty acids. PMID- 9357109 TI - [The 37 annual meeting of the Japanese Society of Nuclear Medicine. Omiya City, Japan. November 19-21, 1997. Abstracts]. PMID- 9357110 TI - [44th meeting of the Japanese Society of Clinical Pathology. Kobe, Japan. October 30-November 1, 1997. Abstracts]. PMID- 9357111 TI - [The 39th congress of the Japanese Society of Clinical Hematology. Tokyo, Japan. October 27-29, 1997. Abstracts]. PMID- 9357112 TI - Education and the cabinet of curiosities at the Francke Foundations. PMID- 9357113 TI - The traffic in Halle Orphanage medications: medicinals, philanthropy, and colonial mission. PMID- 9357115 TI - How shall I take my medicine? Dosages and other matters in eighteenth-century medicine. PMID- 9357114 TI - Academic and practical medicine in Halle during the era of Stahl, Hoffmann, and Juncker. PMID- 9357116 TI - Illness and therapy in two eighteenth-century physician texts. PMID- 9357118 TI - Gadolinium can cool, man. PMID- 9357117 TI - The traffic in medical ideas: popular medical texts as German imports and American imprints. PMID- 9357119 TI - Tobacco: the Third World war. Advice from General Sun Tzu. PMID- 9357120 TI - Research to policy decisions. PMID- 9357121 TI - Attitudes towards genetic counselling and testing among medical students and newly qualified doctors. AB - OBJECTIVES: To determine knowledge about four genetic disorders (Down's syndrome (DS), haemophilia (haem), spinal muscular atrophy type 1 (SMA1) and Huntington's disease (HD)), attitudes towards counselling, acceptability of prenatal diagnosis and termination of pregnancies affected with these conditions. DESIGN: Questionnaire survey of a cohort of medical students and newly qualified doctors. SETTING: Faculty of Medicine, University of Ruhuna. RESULTS: 227 completed questionnaires (111 fourth year and 86 final year students, and 30 demonstrators) were analysed. Awareness of DS and haem, was higher than of SMA1 and HD, and was highest among the demonstrators. Over 80% of the cohort would not counsel directively about future pregnancies and would discuss the diseases with the family or at risk individuals. Prenatal diagnosis was found acceptable for DS, haem and SMA1 by a majority of the cohort. Attitudes to termination of affected pregnancies varied, 88%, 77%, 55% and 36% finding it acceptable for DS, SMA1, haem, and HD respectively, provided legal terminations were available and termination was requested by parents. CONCLUSIONS: This cohort of students and doctors appear to accept the principles of clinical genetics involving non directive counselling, prenatal diagnosis and in some disorders, termination of pregnancy. PMID- 9357122 TI - An audit of structure, process and outcome of care of the diabetic clinic, National Hospital of Sri Lanka. AB - AIMS: To audit the structure, process and outcome of care. SETTING: The diabetic clinic, National of Hospital Sri Lanka (NHSL). METHODS: A previously validated MCQ paper of 10 questions which assessed knowledge of diabetes on insulin therapy, dietary management, management during acute illness and management of emergencies was administered to all patients. The function of the clinic was assessed using previously validated audit case record forms. MEASURES OF OUTCOME: Diabetes knowledge among patients, waiting times, bypassing of local institutions, availability of diagnostic equipment, screening activities and time spent for consultation. RESULTS: The clinic had a daily average attendance of 186 patients seen between 0800 to 1200 hours. A single medical officer spent 2.1 minutes for each patient. No screening was performed. There were no facilities to examine patients or for them to sit during consultation. The diabetes knowledge score was 15.1 (SD 3) from a maximum score of 40.43% had bypassed a local institution. Reasons for bypass included non-availability of drugs and the expectation of quality care at NHSL. Patients spent a mean of 1.5 (SD 0.7) hours travelling to the clinic and waited a mean of 1.56 (SD 0.4) hours to see the doctor and 1.3, (SD 0.12) hours to obtain drugs. CONCLUSIONS: The services of the diabetic clinic do not meet the standards expected of a clinic at a tertiary referral centre. Lack of planning and resources (space, manpower and management skills) can be identified as principal shortcomings. PMID- 9357123 TI - Patient mixed biphasic insulin in a diabetic clinic. AB - INTRODUCTION: Improved glycaemic control is possible with the use of multiple injections of premixed insulin. These are expensive, and not available in state hospitals. OBJECTIVES: To study the cost, patient acceptance and efficacy of a patient mixed and administered combination of soluble and lente (biphasic) insulin administered twice a day. PATIENTS: A cohort of 25 patients with poor glycaemic control on a single dose of 100 units or more of lente insulin. 25 patients matched for age and glycaemic control were used as a control. SETTING: The diabetic clinic of the National Hospital Sri Lanka. METHOD: A prospective study of a cohort of patients. RESULTS: Mean fasting blood glucose decreased from 8.3 mmol/l (SD 3.1) to 6.9 mmol/l (SD 2.3, p < 0.01) and mean blood glucose levels declined from 12.3 mmol/l (SD 4.1) to 10.1 mmol/l (SD 4.7, p < 0.01) in the biphasic group. Total mean insulin dose fell from 80 units (SD 12) to 61 units (SD 11) in the biphasic group, but increased in the control group from 82 units (SD 16) to 91 units (SD 13.1). The diabetes well-being score in the biphasic group was 91.5 (SD 35.3), while the control group had a score of 63.7 (SD 21.3 p < 0.01). Mean glycosylated haemoglobin (HbA1c %) was 8.1 (SD 2.7) in the biphasic group compared to 9.2 (SD 3.3) in the control group. CONCLUSION: Patient mixed and administered biphasic insulin on a twice daily basis is feasible, acceptable to patients, results in better glycaemic control and affords better patient satisfaction. PMID- 9357124 TI - Tuberculous meningitis. PMID- 9357125 TI - Rubella immunisation and pregnancy. PMID- 9357126 TI - Bubonic plague--a clinical case with typical microscopic morphology. PMID- 9357127 TI - Is it necessary to give rabies post-exposure treatment after rodent (rats, mice, squirrels and bandicoots) bites? PMID- 9357128 TI - An audit of admissions to an intensive care unit in a regional centre in Sri Lanka. PMID- 9357129 TI - Sinus histiocytosis with massive lymphadenopathy. PMID- 9357130 TI - Compression neuropathies, including carpal tunnel syndrome. PMID- 9357131 TI - Parainfluenza virus infections in England and Wales. PMID- 9357132 TI - Changes in the incidence of AIDS and in AIDS deaths: the effects of antiretroviral treatment. PMID- 9357133 TI - Seasonal rise in whooping cough. PMID- 9357134 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 9357135 TI - Complement in human disease. PMID- 9357136 TI - Biosensors in immunology: the story so far. AB - In conclusion, biosensors are versatile tools with a range of applications. With a thorough knowledge of possible artefacts and limitations, it possible to perform assays that were heretofore not practicable in immunology. PMID- 9357137 TI - Switching gears during T-cell maturation: RANTES and late transcription. AB - Although much is understood about the induction of genes expressed early (within 24 h) after T-cell activation, little is known about the regulation of expression of genes expressed 'late' (three or more days) post-stimulation. A better understanding of transcriptional regulation at this important stage of T-cell maturation may yield new insights into T-cell development and new immunotherapeutic targets. PMID- 9357138 TI - Speed congenics: a classic technique in the fast lane (relatively speaking). AB - Marker-assisted selection protocol (MASP)-based strategies produce congenic strains with the target gene contained on clearly defined donor-derived genomic intervals in less than half the member of generations required by the classic protocol. Thus, the quality and speed of congenic strain construction are enhanced by this methodology. Here, Edward Wakeland and colleagues compare various MASP-based strategies and discuss their advantages with reference to immunological traits. PMID- 9357139 TI - Th1-type immunity is incompatible with successful pregnancy. AB - Several years ago, the rather provocative question was raised: is successful pregnancy a T helper 2 (Th2) phenomenon? Implicit in this argument is the corollary that unsuccessful pregnancy is a Th1-type situation. Here, evidence from murine and human pregnancy is presented to show that, since Th1-type cytokines mediate pregnancy loss, a shift towards Th1-type immunity may help resolve 'unexplained' pregnancy failure. PMID- 9357140 TI - Protective genes expressed in endothelial cells: a regulatory response to injury. AB - Endothelial cells (ECs) have evolved to guard against insults that incite inflammation. Response to injury is an active process that, if uncontrolled, can progress to EC death (apoptosis). Here Fritz Bach and colleagues suggest that ECs have a balancing component to their proinflammatory response: they upregulate a set of protective genes, including anti-apoptotic genes, that serve to limit the activation process and thereby regulate the response to injury. PMID- 9357142 TI - The host-tumor immune conflict: from immunosuppression to resistance and destruction. AB - A successful immune response against a tumor is dependent on the cytokine repertoire present at the tumor site. Salem Chouaib and colleagues discuss evidence that, to escape the immune system, tumor cells not only produce immunosuppressive cytokines but also employ strategies involving altered susceptibility to tumor necrosis factor and Fas cytotoxic pathways and, in some circumstances, use of the Fas ligand to neutralize effector cells. PMID- 9357141 TI - Taming TNF: strategies to restrain this proinflammatory cytokine. AB - Recent studies have demonstrated the essential role of tumor necrosis factor alpha (TNF-alpha) in rheumatoid arthritis and Crohn's disease. This article discusses agents known to suppress the formation or activity of TNF-alpha, and summarizes clinical studies using anti-TNF-alpha antibodies. PMID- 9357143 TI - CD6-ligand interactions: a paradigm for SRCR domain function? AB - The scavenger receptor cysteine-rich (SRCR) superfamily, which includes proteins expressed by leukocytes, can be subdivided into groups A and B. Group B contains the lymphocyte cell-surface receptor CD6. This article reviews recent progress in understanding the interaction between CD6 and its ligand, activated leukocyte cell adhesion molecule (ALCAM). Analysis of the CD6-ALCAM interaction may help to understand how other SRCR domains bind to their ligands. PMID- 9357144 TI - B-1/macrophages as 'living fossils'. PMID- 9357145 TI - Blind T-cell homeostasis and HIV infection. PMID- 9357146 TI - Th-cell modulation in multiple sclerosis. PMID- 9357148 TI - A placebo controlled trial of two dosages of LPC antagonist--choline in the management of bronchial asthma. AB - The present study was conducted to compare choline with a placebo for its dose effect relationship in management of bronchial asthma. Three groups: Group A: taking placebo (sorbitol), Group B: low dose choline (500 mg TDS), and Group C: high dose choline (1000 mg TDS) were assessed after a trial of four months. A decrease in symptoms score and increase in percent asymptomatic days was observed in all the three groups but was statistically significant only in the groups taking choline (Group B and C). Further, it was more significant in the group taking higher dose of choline (Group C). Average drug-requirement decreased in all the three groups but was significant only in the high dose choline group (Group C) Specific airway conductance (SGaw) at FRC and RV improved significantly only in group taking higher dose of choline (Group C), while no significant change in percent fall in SGaw at RV was observed in any group. It is, thus, concluded that choline is a useful prophylactic drug in the management of bronchial asthma. The improvement was more significant at a higher dose. Further studies are required to establish optimal dose of choline in the management of bronchial asthma. PMID- 9357149 TI - Study of oxygen saturation, breathing pattern and arrhythmias in patients of interstitial lung disease during sleep. AB - Twenty patients of interstitial lung disease (ILD) and same number of healthy adults were selected to monitor arterial hemoglobin oxygen saturation (SaO2) breathing pattern and arrhythmias during sleep. The maximum fall in SaO2 during sleep was 13.1% (10-16%) in ILD patients as compared to 4.8% (3-6%) in controls and the difference was significant (p < 0.005). The ILD patients spent 16.9% of mean total sleep time (TST) below 85% SaO2 and 0.7% of mean TST below 80% SaO2 whereas none of the healthy subjects had SaO2 below 90% during sleep. These patients had more disturbed sleep than controls. Abnormally high breathing frequency demonstrated by ILD patients while awake, was not altered during sleep. Both tidal volume (VT) and minute ventilation (Vmin) decreased by 6.6% and 11.5%, respectively in ILD patients during sleep though it was not significant (p > 0.25) statistically. The respiratory drive was declined during sleep in ILD patients. The percent of tidal volume contributed by rib cage (% RC) lessened during sleep in all the subjects. The ratio of the total excursion of the rib cage and abdomen during inspiration without considering the direction of movement, divided by tidal volume (TCD/VT) revealed asynchronous breathing in ILD patients during sleep. Arrhythmias were found in 6 (30%) of ILD patients and 4 (20%) of control subjects. Observed apnea-hypopnea did not qualify for sleep apnea syndrome in any case. PMID- 9357150 TI - Pulmonary functions in healthy females of Haryana. AB - Pulmonary functions including lung volumes and pulmonary diffusing capacity were assessed in 137 healthy female subjects, 18-52 years of age. The results were analysed by age-wise division of subjects in six groups. It was observed that there were wide variations in pulmonary function values from one individual to another depending upon age and height of the subjects. Majority of ventilatory functions attained maximum values by the age of 25 years while diffusion functions improved upto 35 years of age. Pulmonary functions showed better correlation with age and height as compared to weight. Taking into consideration age and height of subjects, regression equations for different pulmonary functions have been derived for female subjects. PMID- 9357147 TI - Intersubject variability of serum theophylline concentration after single dose of uncoated controlled release theophylline tablet administration in asthmatic patients. AB - Various reports indicate significant intersubject variation in the rate of absorption of controlled release theophylline products that are commercially available. Serum theophylline concentration was estimated in 17 asthma patients treated with controlled release theophylline tablet (mean dose: 5.6 +/- 0.6 mg/kg, mean body wt: 49.82 +/- 9.05 kg). The mean maximal concentration (Cmax) of 5.57 +/- 1.65 micrograms/ml occurred at mean time of maximal concentration (Tmax) of 3.3 +/- 1.36 (mean +/- SD) hours. Mean minimal concentration (Cmin) was 3.164 +/- 1.1 micrograms/ml (mean +/- SD) at mean time of minimal concentration (Tmin) of 5.77 +/- 1.77 (mean +/- SD) hours. The mean percent fluctuation was 112.95 +/- 183.45 (mean +/- SD) but in 4 out of 17 patients, percent fluctuation was 107.29 to 823.10. Therefore, 23.5% patients treated with controlled release theophylline tablet had wide fluctuations in serum theophylline concentration. PMID- 9357151 TI - Pulmonary sequestration: the setting sun sign. PMID- 9357152 TI - Infantile mediastinal primary endodermal sinus tumour. AB - Primary extragonadal mediastinal endodermal sinus tumour is rare, and to date very few cases have been reported in the literature. We present here a case of rare extragonadal highly malignant commonest germ cell tumour in an infant who presented with a rapidly progressive mediastinal mass with dry non-productive cough, tachypnea without significant respiratory distress or toxicity. PMID- 9357153 TI - Multiple neoplasms of the salivary gland and the lung. AB - We describe a patient who presented with Warthin's tumour of the salivary gland which was followed by a bronchial carcinoid and subsequently by a squamous cell carcinoma of the lung. Such a presentation of three neoplasms in an individual is an uncommon occurrence. PMID- 9357154 TI - Constrictive bronchiolitis: a CT scan appearance. AB - Bronchiolitis obliterans or constrictive bronchiolitis is a distant clinical entity with obstructive airway disease. High resolution computed tomography (HRCT) of the thorax can often suggest the diagnosis with the characteristic "mosaic pattern". We present here two cases of bronchiolitis obliterans in which the HRCT showed the presence of this characteristic appearance. PMID- 9357155 TI - Castleman's disease mimicking neoplasia: a case report with review of literature. AB - Castleman's disease is an unusual entity which may at times mimic malignancy but is entirely benign in nature. It is topical as it has been noted to occur with AIDS and Kaposi's sarcoma. We report an interesting case with a mediastinal mass which was thought to be neoplastic but turned out to be Castleman's disease on histopathology. PMID- 9357156 TI - Clot lysis: role of plasminogen activator inhibitors in haemostasis and therapy. AB - An unimpeded circulation of blood depends on the concerted activation of coagulation and fibrinolytic factors. The latter entails the controlled, localised conversion of plasma zymogen plasminogen to the active enzyme plasmin mediated by tissue-type plasminogen activator (tPA). Bulk of tPA activity is in the proximity of the endogenous plasminogen activator inhibitor (PAI) as an active complex. The advent of molecular biology techniques has enabled isolation of cDNA for the inhibitors PAI-1, PAI-2 and PAI-3 and data indicate that these belong to the serine protease inhibitor (Serpine) family with arginine as its active site but immunologically distinct from each other. Enhanced tPA or PAI-1 forms one of the risk factors related to cardiac diseases and thrombotic disorders. A line of therapy entails lowering of PAIs with concomitant increase in tPA levels leading to net enhancement in fibrinolytic activity. In as much as plasminogen activators exert their action extracellularly, they are accessible to inhibitors and therefore PAIs could have a therapeutic potential and serve as prognostic indicators in cancer. Documented findings related to the biochemical characteristics and therapeutic potential of PAIs are presented and discussed in the review. PMID- 9357157 TI - Role of proteases in tumor invasion and metastasis. AB - Cancer remains a major cause of worldwide deaths due to ability of cancer cells to form secondary tumors at other sites by multistep process called metastasis. In order to migrate from their original site, tumor cells have to cross several barriers like basement membranes, interstitial tissues and extracellular matrices, which are composed primarily of collagen, proteoglycans, elastin, laminin and other glycoproteins. Tumor cells over express and secrete proteases which are capable of degrading the components of these barriers and thus facilitate their migration. The classes of proteases which have been implicated in the process of tumor invasion and metastasis include metalloproteases, serine proteases and cathepsins. Cancer cells in general have elevated levels of proteases belonging to more than one class. In some studies, process of invasion has been inhibited by using specific inhibitors of these proteases. Expression of some proteases has been observed only in some specific tumors. These proteases have been proposed to be of diagnostic/prognostic value. However a better understanding of the process of metastasis and tumor invasion is required before proteases can be used as therapeutic targets for blocking the spread of cancer. PMID- 9357158 TI - Experimental methods for evaluation of psychotropic agents in rodents: I--Anti anxiety agents. AB - Rodent models of clinical anxiety are extensively used for evaluating putative anxiolytic activity. In the present review, the available methods which can be utilized by most laboratories, have been discussed. These methods have been categorized as methods involving conditioning techniques and those not involving conditioning. In most cases, the methodology has been briefly discussed in terms of experimental use and efficacy of benzodiazepine and the newer non benzodiazepine anxiolytics. PMID- 9357159 TI - Effect of vitamin A excess on germ cell development in prepubertal rat testis. AB - Vitamin A in graded doses of 125, 250 and 375 U.S.P./kg body wt, po, for 10 days (d 21-30) drastically reduced the testicular weight by 25 to 62% and seminiferous tubular diameter by 14 to 35% in prepubertal rats in lowest and highest doses of the treatment. The treatment induced disproportionate enlargement of nuclei and cytoplasm of the germ cells; predominantly the preleptotene and pachytene spermatocytes. These abnormal germ cells, often with 2 or 3 nuclei displayed vacuolated cytoplasm surrounding pyknotic or granulated or dispersed chromatin granules within the nuclei in a dose proportionate manner. The round spermatids were the most sensitive cell types which completely disappeared in two higher doses of treatment. Vacuolation of Sertoli cell cytoplasm in about 25% of the tubules with associated increase in intertubular space was also observed in rats treated with the highest dose of the vitamin. Circulatory levels of FSH, LH and testosterone remained unaltered following the vitamin excess treatment. Therefore, it is suggested that excess vitamin A even for shorter duration like the present one is detrimental to developing cell types and prevents the progress of the spermatogenic process beyond the round spermatid stage. PMID- 9357161 TI - Relative potentiality of some contragestational agents on trauma induced deciduoma formation in immature rats. AB - Relative progestational and antiprogestational profile of four contragestational agents, viz. RMI-12,936, RU-38,486, STS-557 and WIN-32,729, known to possess either or both the properties in other test systems, was determined in immature rats using deciduoma induction as marker. The deciduoma was produced by needle traumatization in one of the uterine horn, and the extent of stimulation was analysed, both macro and microscopically, against the control nontraumatized horn. The results indicated that the agents which demonstrated better potentiality(ies) as progestational and/or antiprogestational in this bioassay exhibit contraceptive efficacy at relatively lower doses as compared to those that showed mild to moderate potentiality. PMID- 9357160 TI - Adoptive immunotherapy of murine lymphosarcoma: comparative study using allo immune or tumor-immune T cells. AB - The antitumor effect of allosensitization with lymphocytes and skin graft of DBA/2 mice was evaluated using immunogeneic, transplantable Lymphosarcoma (LS-A) syngeneic to Swiss mice. A dose dependent tumor inhibitory effect in terms of tumor free mice was observed in mice sensitized i.p. with lymphocyte doses between 10-100 million per animal. Sensitization with allogeneic primary skin graft was more effective than lymphocyte immunization. The antitumor immunity could be adoptively transferred in syngeneic Swiss mice using either allo-immune or tumor-immune T cells. Analysis of T cell phenotypes using monoclonal antibodies against cell surface markers CD4 and CD8, indicated absolute dependence on the CD4+ T cells subset in tumor cure in case of allo-immune as well as tumor-immune T cells. CD8+ T cell subset was found essential only in case of allo-immune T cell therapy. Immunosuppression of mice with whole body gamma irradiation (4 Gy), 6 hr before transfer of allo-immune or tumor-immune T cells did not abrogate the therapeutic ability of allo-immune or tumor-immune T cells. Our results suggest that allosensitization could be an effective method of generating effector lymphocyte populations that might be used to treat tumors that exhibit detectable immunogenecity. PMID- 9357162 TI - Effect of pinealectomy and testosterone on gonadal regression and accessory sex organs in Indian palm squirrel, Funambulus pennanti. AB - Indian palm squirrel (Funambulus pennanti), is a tropical seasonal breeder presents a short gonadal regression period (October-November), concomitant with declining plasma testosterone level. If pinealectomized (Px), however, squirrels maintain full gonadal activity and plasma testosterone levels. The aim of the present study was, therefore, to determine if the Px and exogenous testosterone in intact and Px squirrels would prevent natural gonadal regression. Experiment was performed during testicular regression phase (i.e. August end to November) revealed that both the Px (60 days) and exogenous testosterone (100 micrograms/day/squirrel/30 days) prevented gonadal and accessory sex organ regression. This could be due to negative feedback by testosterone treatment which was more obvious in intact than the Px squirrels. A decline in day time plasma melatonin level after testosterone treatment was also indicative of inverse relationship between the gonadal and pineal hormones which in turn did not allow the collapse of accessory sex organs. Further, the pineal gland is under direct control of steroid hormones since receptors and aromatizing enzymes are noted in the same. Therefore, a pineal secreted melatonin stimulation of hypothalamic negative feedback centers for gonadal regression could be suggested in this tropical rodent as receptors for melatonin have been detected in above areas. PMID- 9357163 TI - Isolation and characterization of soluble antigens of sheep poxvirus. AB - A soluble antigen fraction of sheep poxvirus (SPV) isolated from infectious virus particles by ultracentrifugation and purified by subtractive immunoaffinity chromatography was characterized. Exclusion chromatography studies revealed 10 proteins of molecular weight (MW) 220, 168, 87.3, 71.5, 52.5, 36.7, 31.0, 23.4, 18.3 and 14.2 kDa. Nine of them were found to be precipitinogens and 5 were identified as structural components of the virus particles. SDS-PAGE analysis revealed a polypeptide profile of 10 bands with 2 prominent polypeptides of 64 and 42 kDa. Western blotting, however, detected 2 immunogenic polypeptides of MW 100 and 64 kDa. Moreover, crossed immunoelectrophoresis showed the presence of proteins of varied electrophoretic mobility and sharing of antigenic determinants among a few soluble antigens. Physico-chemical characterization further revealed that these precipitinogens can withstand ambient temperatures, but were sensitive to trypsin and ether whereas, chloroform had no effect on immunoprecipitation pattern of soluble antigens. PMID- 9357164 TI - Differential biochemical response of freshly isolated rat hepatocytes to paracetamol, carbon tetrachloride and D-galactosamine toxicity. AB - Differential response of freshly isolated rat hepatocytes to paracetamol (acetaminophen, AAP), carbon tetrachloride (CCl4) and D-galactosamine (GalN) was examined. The viability of the cells in suspension culture was not altered for 4 hr when incubated in Hank's balanced salt solution (HBSS) supplemented with 4.2 mM NaHCO3, 10 mM HEPES buffer and 0.5% bovine serum albumin. AAP induced time and dose dependent depletion of GSH as an early manifestation of AAP toxicity. Hepatocytes exposed to AAP exhibited lower lactate dehydrogenase (LDH) activity released into the medium than in controls. This was due to the interaction of a reactive metabolite of AAP, i. e. N-acetyl p-benzoquinoneimine (NAPQI) with cell proteins. Hepatocytes isolated from rats pretreated with 3-methylcholanthrene expressed higher sensitivity to AAP toxicity at least by a factor of 5. Furthermore, AAP-induced toxicity was not found to be related to any lipoperoxidative stress. CCl4 on the other hand elicited a highly lipoperoxidative response in hepatocytes and consequent leakage of cellular enzymes with lengths of incubation. Again the sensitivity of the response to CCl4 was enhanced remarkably in hepatocytes isolated from phenobarbital- pretreated rats; LDH leakage increased by 3-fold and thio-barbituric acid reactive substances by 25-fold. Unlike the two toxicants, galactosamine depleted UDP glucuronic acid in a concentration and time-related manners. The differential biochemical response of hepatocytes to three hepatotoxicants investigated may prove useful as rapid in vitro screen for selection of compounds of hepatoprotective potentials. PMID- 9357165 TI - Silymarin mediated differential modulation of toxicity induced by carbon tetrachloride, paracetamol and D-galactosamine in freshly isolated rat hepatocytes. AB - Influence of silymarin on the modulation of hepatotoxicity induced by carbon tetrachloride (CCl4), paracetamol (AAP) and D-galactosamine (GalN) was examined in freshly isolated rat hepatocytes in suspension culture. While the three hepatotoxicants produced differential biochemical response, the flavone was able to restore biochemical alterations only in hepatocytes exposed to CCl4 and AAP induced toxicity. Silymarin at 0.4 mM was able to counteract lipid peroxidation and enzyme leakage induced by 3 mM CCl4 The flavone also offered protection by more than 60% in hepatocytes isolated from PB pre-treated rats where CCl4 at 2 mM produced enhanced toxicity over hepatocytes isolated from untreated control rats. Similarly, the flavone protected AAP-induced GSH depletion by more than 75% in hepatocytes isolated from untreated and 3-methylcholanthrene treated rats. However, instead of protecting GalN-induced depletion of UDP-glucuronic acid in hepatocytes, the flavone itself reduced the nucleotide content very rapidly compared to GalN, the later exerted time dependent effect. Silymarin at 0.4 mM reduced UDPGA by more than 60%. The results suggested that freshly isolated hepatocytes in suspension culture offer a simple and convenient method for evaluation of pharmaceutical agents of antihepatotxic potentials against various hepatotoxicants. PMID- 9357166 TI - Dose-dependent response of central dopaminergic systems to metoclopramide in mice. AB - Metoclopramide (5 to 40 mg/kg, i.p.) induces catalepsy and antagonised apomorphine induced cage climbing behaviour in mice. This further indicate its postsynaptic striatal and mesolimbic D 2 dopamine (DA) receptor blocking activity. Metoclopramide at 1.25 and 2.5 mg/kg, i.p. induced stereotyped cage climbing behaviour in mice. Pretreatment with haloperidol and alpha-methyl-p tyrosine significantly antagonised metoclopramide (1.25 and 2.5 mg/kg)-induced stereotyped cage climbing behaviour. Metoclopramide at these doses induces stereotyped cage climbing behaviour by releasing DA from the mesolimbic dopaminergic neurons with resultant activation of the postsynaptic mesolimbic D 2 DA receptors by the released DA. DA releasing action of metoclopramide (1.25 and 2.5 mg/kg, i.p.) and the subsequent induction of the stereotyped cage climbing behaviour by these doses of metoclopramide is explained on the basis of selective blockade of the presynaptic D 2 DA autoreceptors by these doses of metoclopramide. PMID- 9357167 TI - Neurobehavioral effects of low level fenvalerate exposure in mice. AB - Effect of oral administration of fenvalerate, a pyrethroid insecticide was studied in different behavioral paradigms in mice. Fenvalerate at 15, 30 and 45 mg/kg dose increased start latency, decreased ambulation and rearing in open field, increased immobility in tail-suspension test and attenuated haloperidol induced catalepsy in a dose-dependent manner. The time-course of data shows that these effects of fenvalerate may sustain up to several hours of its oral administration. The study indicates that pyrethroids can cause adverse behavioral effects even after a very low-level exposure. Although, it is difficult to extrapolate these findings directly to behavioral changes in man, they indicate that subtle behavioral dysfunction also occur in humans at exposures which do not cause acute toxicity. PMID- 9357168 TI - Interspecific hybridization in poeciliids. AB - Seventy per cent attempts to ensure interspecific hybridization between Poecilia velifera and P. sphenops were successful and led to the production of true hybrids, but not gynogens or triploids, as evidenced by the mottled or striped colour, chromosome number (2n = 46) and response from scale transplantation. Most hyrbids were infertile as they failed to cross among themselves or with their respective parents; however they were more closely related to P. sphenops as indicated by mating responses and scale transplantation studies. Heterospecific impregnation resulted in 40% reduction in fecundity but retention of interparturition period characteristic of the female species. A skewed ratio of 3 Female Female:2 Male Male, observed in the laboratory populations of both P. velifera and P. sphenops, was also sustained among the progenies sired from heterospecifically inseminated females. The colour patterns of hybrids were of two types: mottled and striped, the latter one being reported for the first time. PMID- 9357169 TI - Bioactivity of marine organisms: Part VIII--Screening of some marine flora of western coast of India. AB - Methanolic extracts of 31 botanically identified species of marine flora, collected from Gujarat Coast, have been screened for a wide range of biological activities. Of these, 3 extracts showed anti-implantation, 2 had antiviral, 2 showed hypotensive, 1 had anti-inflammatory while 12 extracts showed diuretic activities. The antiviral activity; against EMCV, was confirmed in one alga. The active principles and results of these studies are reported. PMID- 9357170 TI - Identification and characterization of some PR-proteins induced by kitazin and Rhizoctonia solani causing sheath blight of rice. AB - Significant reduction (68.38%) in sheath blight disease of rice was noticed when foliar spray of a systemic fungicide, kitazin (480 micrograms mg-1), was applied twice at an interval of 2 days before inoculation. SDS-PAGE analysis of proteins of Rhizoctonia-infected rice leaf sheaths revealed the presence of 16 proteins ranging from 20 to 90 kDa (approx.). Six were identified as constitutive defense proteins (increased after infection), 6 as secondary defense proteins (formed de novo) and the rest 4 appeared non-defense proteins. Non-inoculated kitazin treated leaf sheaths showed 15 proteins of which 5 were constitutive and 4 secondary defense proteins (both are PR-proteins). Among the PR-proteins, five beta-1,3-glucanases and one chitinase was identified and characterized. One rice chitinase (MW 20 kDa) and 2 glucanases (60 & 69 kDa) showed serological relationships with tobacco chitinase (32 kDa) and tobacco glucanase (33 kDa) respectively. The implications of results have been discussed in relation to biotic and abiotic induction of PR-proteins in rice. PMID- 9357171 TI - Mechanism of cardiotoxicity induced by a marine toxin isolated from Ptychodiscus brevis. AB - An organophosphate toxin of marine origin isolated from red tide dinoflagellate P. brevis produced a dose-dependent dual effect on rat atria, i.e. positive inotropic effect at low concentrations (2.8 x 10(-8) to 8.4 x 10(-7) M) and negative inotropic and chronotropic responses at an elevated dose (4.8 x 10(-6) to 7.2 x 10(-4) M). The negative chronotropic and inotropic responses of the toxin were potentiated with physostigmine and ouabain whereas antagonized by atropine and hemicholinium-3 pretreatments and those effects remained unaltered by isoproterenol, phenylephrine and ouabain pretreatments. The results indicate that the toxin induced negative inotropic and chronotropic effects are mediated through release of acetylcholine from the nerve endings and consequent activation of muscarinic receptor. In atria exposed to guanethidine, bretylium, propranolol and tyramine tachyphylaxis, the positive inotropic response of the toxin was not modified. However, the response was antagonized by EGTA, nifedipine, ryanodine, calcium-free ringer and potentiated with caffeine and amiloride pretreatments. The results suggest that the positive inotropic effect of the toxin is mediated through Ca2+ influx and impairment of Na+/Ca2+ exchange process. PMID- 9357172 TI - Hepatoprotective effect of Liv-52 and kumaryasava on carbon tetrachloride induced hepatic damage in rats. AB - Oral administration of Liv-52 and kumaryasava to carbon tetrachloride (CCl4) treated rats improved growth. Kumaryasava was more effective in reducing the liver weight increase due to hepatotoxicity of CCl4. Hepatic arginase, cathepsin B, acid phosphatase, ribonuclease activity which were decreased on CCl4 treatment was stimulated by both Liv-52 and kumaryasava. Results indicate that Liv-52 and kumaryasava have protective effect on hepatic enzyme induced due to CCl4 hepatotoxicity. PMID- 9357173 TI - Genotoxic effects of poly-D-lysine in m-AMSA (amsacrine) treated Chinese hamster ovary (CHO) cells. AB - Cultured Chinese hamster ovary (CHO) cells were pre-treated with m-AMSA (amsacrine) and post-treated with different doses of polycationic compound poly-D lysine (PDL) during G2 period in order to test on the frequency of chromatid-type aberrations. The present results indicate that PDL enhances the genotoxic action of m-AMSA measured as induced chromatid aberrations. PMID- 9357174 TI - Glutathione status of some homeothermic vertebrate species and its relation with diabetogenesis. AB - The pancreatic glutathione levels of birds are higher than that of mammalian species. Refractoriness of birds towards the action of diabetogens may indeed be correlated with this fact. However, the higher pancreatic glutathione value found in owl, in comparison to that of pigeon, seems contradictory since owl is more susceptible to diabetogens. PMID- 9357175 TI - An electronic device to record behavioural activity of bivalves. AB - An electronic device to record the valvular movements of clams was fabricated using a Hall effect transducer. It was used to record the responses of Anadara granosa, an arcid clam harvested from coastal waters of Bombay, to a chemical toxicant (10 ppm CuSO4) after 96 hr exposure to naphthalene (5, 10 and 15 ppm). The clams exposed to naphthalene did not respond to the presence of a chemical toxicant (10 ppm CuSO4) while the control clams responded and closed their shells rapidly. PMID- 9357176 TI - Response of some Calvin cycle enzymes subjected to salinity shock in vitro. AB - Leaf enzyme extracts of 10 day old seedlings of horsegram were subjected to NaCl or Na2SO4 treatment in vitro. Salinity shock caused decline in the activities of RuBP carboxylase, R-5-P kinase, R-5-P isomerase and NADP-Gly-3-P dehydrogenase. At low concentrations, Na2SO4 did not alter the activities of R-5-P kinase, R-5-P isomerase and NADP-Gly-3-P dehydrogenase. RuBP carboxylase was found to be more sensitive to salt shock than the other enzymes studied. Further, NaCl was more toxic to the enzyme activities as compared to Na2SO4. PMID- 9357177 TI - PCR amplification of ribosomal DNA spacer regions of Salmonella enteritidis isolates. AB - Polymerase chain reaction (PCR) was used to amplify ribosomal DNA spacer regions from nine Salmonella enteritidis field isolates. Unique products of 480 and 660 bp were obtained from the isolates. PCR product (480bp) was then cloned into pUC18 vector by blunt end ligation. PMID- 9357178 TI - Evaluation of different rodenticidal baits against rodent population in cucumber (Cucumis sativus) crop fields. AB - The baits used consisted of 2% zinc phosphide and 0.005% bromadiolone. The baitings were carried out at the flowering stage of cucumber crop. Double baiting treatments comprising zinc phosphide and bromadiolone in different combinations resulted in better reduction of rodent population and crop damage. Deployment of different rodenticidal treatments saved yield loss from 7.00 to 11.82q/ha. Thus, accomplishing cost benefit ratio of 1:88 to 1:107. Bromadiolone followed by bromadiolone, the most profitable combination is suggested for control of rodent population in cucumber crop fields. PMID- 9357179 TI - Antifeedant activity of some new heterocyclic compounds against the larvae of Spodoptera litura F. AB - The antifeedant activity of diisoflavones a synthetic products is reported for the first time against S. litura. The antifeedant activity was tested employing the non-choice test method against the 4th instar pre-starved larvae. PMID- 9357180 TI - Interrelationship between amines level in brain regions of fish, Oreochromis mossambicus during organophosphorus stress. AB - Amines such as dopamine, norepinephrine and epinephrine were analysed in the brain regions of O.mossambicus exposed to quinalphos, phenthoate and their combination for 96 hr. The three types of treatments significantly (P < 0.05) altered the amines level at various intervals in the brain regions. PMID- 9357181 TI - Is it curtains for chronic amebiasis? PMID- 9357182 TI - Does Entamoeba histolytica cause irritable bowel syndrome? AB - BACKGROUND: Symptoms of patients with irritable bowel syndrome (IBS) closely mimic those of patients with non-dysenteric amebic colitis. AIM: To examine the clinical relevance of presence and types of Entamoeba histolytica in stools of patients with IBS. METHODS: IBS was diagnosed by Manning's criteria. Stool examination was done 4-weekly for 48 weeks to detect E. histolytica cysts or trophozoites. Patients underwent initial sigmoidoscopy. Sera of 22 IBS patients, 23 asymptomatic cyst passers and 36 healthy volunteers whose stools were also examined were tested for presence of antiamebic antibodies. Stools were cultured for amebae; positive cultures were subjected to polyacrylamide-gel electrophoresis (PAGE) using hexokinase (HK) isoenzyme to distinguish between pathogenic (fast-moving band) E. histolytica infection and nonpathogenic (slow band) species of Entamoeba dispar. RESULTS: E. histolytica cultured from stool samples of four IBS patients had slow-moving band of HK on PAGE. All patients spontaneously eradicated the infection during the next eight to 24 weeks; all had negative serology for antiamebic antibodies, and normal rectal mucosa on sigmoidoscopy. No change in symptom score occurred on follow up in IBS patients, although all of them cleared the infection. Three additional E. histolytica isolates from IBS patients obtained from another laboratory also showed nonpathogenic isoenzyme pattern. CONCLUSION: Bowel symptoms in IBS patients were not related to E. histolytica infection. The term non-dysenteric amebic colitis thus appears to be inappropriate, since it may be used erroneously for patients with IBS with nonpathogenic ameba, leading to injudicious treatment with antiamebic drugs. PMID- 9357183 TI - Mesenteric vasculopathy in intestinal tuberculosis. AB - BACKGROUND: Involvement of mesenteric vessels in intestinal tuberculosis and its role in the pathogenesis of the intestinal changes have not been studied histologically. AIM: To study mesenteric vessels in patients undergoing surgery for complications of intestinal tuberculosis. METHODS: Resected intestinal specimens from 68 patients presenting with intestinal perforation and intestinal obstruction were examined; involvement of the major mesenteric vessels was evaluated. RESULTS: Granulomas were seen in the vessel wall in one case and near the vessel wall in 11 cases, intraluminal thrombi were seen in 23 cases, and subintimal fibrosis in nine cases. Perivascular cuffing was seen in intramural and subserosal vessels in ten cases. CONCLUSIONS: Changes in the vessel wall may lead to gut ischemia, which may contribute to the development of strictures and stercoral perforation in intestinal tuberculosis. PMID- 9357184 TI - Metronidazole relieves symptoms in irritable bowel syndrome: the confusion with so-called 'chronic amebiasis'. AB - BACKGROUND: Metronidazole is often administered to patients with irritable bowel syndrome with an erroneous diagnosis of 'chronic amebiasis'. AIMS: To assess how patients with irritable bowel syndrome respond to metronidazole in the absence of amebae in their stools. METHODS: We randomly allocated 45 patients (35 men; aged 15-59 years) with irritable bowel syndrome to receive isapghul (10 g bid x 60 days), metronidazole (400 mg tid X 10 days, followed by placebo x 50 days), or placebo (1 capsule bid x 60 days). Symptoms were evaluated and scored on days 0, 15, 30, 45 and 60. Rectosigmoid manometry was performed in 5 of 15 patients in each group on days 0 and 60. RESULTS: There was a significant time effect and treatment effect on the symptom scores in all groups (isapghul > metronidazole > placebo); total score decreased from mean 25.8, 24.0 and 24.6 on day 0 to 7.2, 10.9 and 18.1 on day 60, respectively. Severity, duration and frequency of pain; and mucus in stool were all significantly reduced in all treatment groups (p < 0.001 for each). Treatment with isapghul increased the mean amplitude of propagated activity from 26.2 mmHg to 30.1 mmHg at 20 cm (p < 0.025) and from 23.1 mmHg to 27.4 mmHg at 10 cm (p < 0.05) from the anal verge, as well as the total duration of propagated activity at both sites (p < 0.05), with decrease in number of propagated contractions per 10 min (p < 0.025). Metronidazole and placebo had no effect on manometric findings. CONCLUSIONS: Metronidazole provides symptom relief in irritable bowel syndrome, without affecting rectosigmoid motility. This symptom response may be misinterpreted as supporting a diagnosis of 'chronic amebiasis'. PMID- 9357185 TI - Mucin histochemistry of esophagus in health and malignancy. AB - AIMS: To study mucin histochemistry of the normal esophagus, esophageal adenocarcinoma, and carcinoma exhibiting glandular and squamous elements, to ascertain the origin of these tumors. METHODS: Mucin histochemistry was studied in sections of the normal cardioesophageal junction obtained from 25 post-mortem specimens and in 12 mucin-secreting esophageal carcinomas. RESULTS: The normal submucosal esophageal glands and three adenocarcinomas secreted predominantly sulfomucins; a mixture of neutral and sialomucins was seen in the nine carcinomas with squamous and glandular traits. Barrett's metaplasia was not encountered. CONCLUSIONS: In the absence of Barrett's metaplasia, esophageal adenocarcinoma probably arises from the submucosal glands, whereas squamous carcinomas with mucin-secreting component could arise from metaplastic change in squamous epithelium, cardiac glands, or multipotent stem cells in the epithelium. PMID- 9357186 TI - Transrectal ultrasonography versus computed tomography in staging rectal carcinoma. AB - BACKGROUND: Since therapy of rectal carcinoma depends on the extent of disease, staging becomes important. AIM: To assess the ability of transrectal ultrasonography (TRUS) and computed tomography (CT) to stage rectal carcinoma. METHODS: Ten patients with rectal carcinoma were examined by TRUS and plain and contrast-enhanced CT scan; their findings were compared with each other and with those at surgery. RESULTS: TRUS identified wall invasion in all ten cases and perirectal fat infiltration in all five cases in whom these were present. Node involvement was detected in five cases on TRUS and two of six cases on CT. Metastasis to bladder (one case) was not recognized by TRUS but was seen on CT. CONCLUSION: TRUS is inexpensive and superior to CT in staging early rectal carcinoma; limited depth of penetration is its major limitation. CT is useful for the diagnosis of advanced disease. PMID- 9357187 TI - Liver resection. PMID- 9357188 TI - Combined percutaneous-endoscopic approach for biliary endoprosthesis placement. AB - A nonoperative method of palliation was used in four patients with malignant obstructive jaundice in whom biliary endoprosthesis could not be placed endoscopically. A guide wire was manipulated through the lesion by a percutaneous transhepatic route and retrieved from the duodenum through an endoscope. A 10 Fr stent was then passed through the endoscope over the guide wire across the stricture. The procedure was successful in all four patients, with no complication. PMID- 9357189 TI - Endoscopic management of pancreatic diseases. AB - Endoscopic management has recently been used for a variety of chronic pancreatic diseases. We used this approach in five patients with pancreatic diseases (calcific pancreatitis 2, pancreatic pseudocyst 3). Nasocystic drain was placed in a patient with pancreatic pseudocyst at the tail end of the pancreas; a 5 Fr stent was placed over 0.021"/0.035" guide wire in the main pancreatic duct in the others. All patients had relief of pain. Nasocystic drain led to resolution of pseudocyst, perisplenic collection and pleural effusion. Endoscopic treatment is safe and effective in various pancreatic disorders. PMID- 9357190 TI - Intestinal lymphangiectasia: presentation in pregnancy and association with herpes zoster and alopecia. AB - We report a woman with intestinal lymphangiectasia whose symptoms were wrongly attributed to pregnancy; the diagnosis was made in the postpartum period. She also developed alopecia and herpes zoster. PMID- 9357191 TI - Injection polypectomy of gastric polyps. PMID- 9357192 TI - ERCP in situs inversus: do we need to turn the other way? AB - Endoscopic procedures are difficult in patients with situs inversus owing to left right reversal of viscera. Conventionally, reversal of the position of the endoscopist in relation to the patient is advocated to overcome the anatomical difficulty. We describe a patient with chronic calcific pancreatitis and pseudocyst of the pancreas who had situs inversus. ERCP was performed with minor modification of maneuvers with the patient in the usual left lateral decubitus and the endoscopist on the left of the table. PMID- 9357193 TI - Adult intrahepatic solitary choledochal cyst. AB - A 20-year-old woman presented with dull ache in the abdomen. Ultrasonography and CT scan showed a solitary liver cyst. Roux-en-Y cystojejunostomy was done since there was bile in the cyst. Histology of the cyst wall revealed it to be a choledochal cyst. Solitary intrahepatic choledochal cyst and presentation in adult life are rare. PMID- 9357194 TI - Adenocarcinoma of cervical esophagus arising in aberrant gastric mucosa. AB - Ectopic gastric epithelium is common in the cervical esophagus. However, complications arising from such tissue are rare. We report an adenocarcinoma arising in ectopic gastric mucosa of the cardiac type in the cervical esophagus of a 60-year-old man. PMID- 9357195 TI - Persistent transaminase elevation due to heterozygous (familial) apolipoprotein B deficiency. AB - Homozygous apolipoprotein B deficiency can present with fatty liver and raised levels of transaminases. Subjects with heterozygous deficiency are almost always asymptomatic. We report an asymptomatic 26-year-old man with persistently raised transaminases, in whom the diagnosis of heterozygous (familial) apolipoprotein B deficiency was made on the basis of characteristic lipid profile. PMID- 9357196 TI - Mesenteric lipoma: an unusual cause of small intestinal volvulus. AB - Mesenteric lipoma as a cause of small intestinal volvulus has not been reported before. We report a middle-aged man with this entity. PMID- 9357197 TI - Tracheo-bronchial remnants: a rare cause of dysphagia. AB - A 14-year-old girl presented with dysphagia since the age of one year. Investigations revealed two strictures at the lower esophagus. Endoscopic mucosal biopsy from the strictures showed pseudostratified columnar epithelium suggesting that tracheobronchial remnants was the cause of the stenosis. Esophageal dilatation failed to relieve her symptoms. PMID- 9357198 TI - Seroconversion with rDNA hepatitis B vaccine of Cuban origin. PMID- 9357199 TI - Diagnosis of subclinical intestinal obstruction using hepatobiliary scintigraphy. PMID- 9357200 TI - HBsAg prevalence in blood donors in Jaipur. PMID- 9357202 TI - Prevention of vertical transmission of human immunodeficiency virus. PMID- 9357201 TI - HIV liver disease: the Indian scenario. PMID- 9357203 TI - A clinicopathologic profile of adrenocortical tumors. AB - OBJECTIVE: To study the clinical, biochemical, hormonal, radiological and histopathological profile of adrenocortical tumors in children; to assess the clinicopathological correlations and note the future outcome. DESIGN: Retrospective and prospective study. SETTING: Hospital based; Endocrine Service of our institution and other institution based services. SUBJECTS: 14 children (Females = 11, Males = 3) with adrenocortical tumor, aged 8 months to 13 years (mean age 5.1 +/- 3.42 years), seen over a period of 9 years. RESULTS: Females predominated (F:M = 3.7:1). Majority (64%) had a mixed picture with cushingoid features and virilization, whereas 36% presented only for virilization. Elevated serum cortisol levels with loss of diurnal variation was noted only in those with mixed clinical presentation. Adrenal androgen elevation was noted in majority of cases as virilization was common to all. CT confirmed the diagnosis of tumor, 7 on either side. Thirteen cases were operated. Histopathologic diagnosis was carcinoma in 7 and adenoma in 6 cases. Three of the seven with carcinoma died within 3 months to 2 years but two of these with small tumours (weight 60-65 g and diameter < 6 cm) were well at 2 and 5 years, while as one of the six with a large adenoma had recurrence and metastasis after three years. CONCLUSION: Female preponderance was marked (4 times), 43% of tumors had occurred by 3 years of age and 64% by 6 years. Neither the hormonal parameters nor the histopathology correlated well with the biological behavior and outcome. Prolonged and vigilant follow up is essential. PMID- 9357204 TI - Early predictors of neurodevelopmental outcome in high risk infants. AB - OBJECTIVE: To find out a few simple and easily elicitable items at three and six months of age that can predict neurodevelopmental outcome at one year in high risk babies. DESIGN: One year longitudinal follow up study. SETTING: Hospital based study including inborn and outborn infants discharged from the Neonatal Intensive Care Unit (NICU) of a referral hospital, followed up in a High Risk Clinic. METHODS: Sixty high risk babies were followed up longitudinally for a period of one year. A detailed neurodevelopmental examination was done with special attention to the following items-axillary suspension, head support, social smile, disappearance of primitive reflexes and neurobehavior at three months age while pull to sit, rolling over, sitting momentarily without support, transfer of objects and voluntary reach were evaluated at six months age. Bayley Scales of Infant Development (Baroda Norms) was used for assessing the outcome at one year. RESULTS: Babies with absence of social smile, abnormal neurobehavior at three months and absent pulling to sit position, absent voluntary reach, and absent transfer of objects, remained delayed at one year. The specificity of each of these items was 100%. These items had a positive predictive value of 100%. CONCLUSIONS: Inability to achieve social smile and abnormal neurobehavior at three months age and absence of pulling to sit position, transfer of objects and voluntary reach at six months age, warrant early intervention. These items are easy to elicit, do not require any special kit or elaborate training. Hence these items can be tested even by those working at the primary level or in office practice. PMID- 9357205 TI - Comparative efficacy of jet nebulizer and metered dose inhaler with spacer device in the treatment of acute asthma. AB - OBJECTIVE: To compare the relative efficacy of jet nebulizer and metered dose inhaler (MDI) with spacer for the administration of aerosolized salbutamol in an acute exacerbation of bronchial asthma. DESIGN: Randomized prospective study. SETTING: Emergency Room. METHODS: In 60 subjects with acute asthma aged between 1 to 12 years, clinical and laboratory assessment of severity at recruitment included heart rate, respiratory rate, pulsus paradoxus, arterial blood gas analysis (all cases) and peak expiratory flow rate (wherever possible). The subjects were randomized into two equal groups to receive aerosolized salbutamol either via nebulizer (Group I) or MDI-spacer (Group II) as per the Consensus Guidelines. The response to therapy was sequentially assessed after 20, 40 and 60 minutes of institution of therapy. RESULTS: A significantly (p < 0.02) greater number of subjects in Group II presented with severe dyspnea and intercostal muscle retraction (subjective assessment). However, the objectively evaluable outcome parameters were comparable (p > 0.05) in both groups at presentation. All the outcome measures showed a significant (p < 0.05) improvement with time in both the groups. The recovery parameters were comparable (p > 0.05) at different time periods in the two groups. CONCLUSION: MDI-spacer is as effective as a nebulizer for the aerosolized administration of salbutamol in an acute exacerbation of asthma in children. However, for developing countries, distinct advantages (economic and power requirement) argue strongly for utilization of MDI spacer in preference to nebulizer. PMID- 9357206 TI - Hematopoietic stem cell transplantation. PMID- 9357207 TI - Hepatitis B and breastfeeding. PMID- 9357208 TI - Adreno-cortical tumors: clinico-pathological profile of five cases. PMID- 9357209 TI - Experience with diarrhea training and treatment unit in Shimla. PMID- 9357210 TI - Pulmonary agenesis. PMID- 9357211 TI - Bardet-Biedl syndrome. PMID- 9357212 TI - Osteopenia of prematurity. PMID- 9357213 TI - Primary Fanconi syndrome. PMID- 9357214 TI - Pneumocystis Carinii pneumonia in pediatric acquired immunodeficiency syndrome. PMID- 9357215 TI - Measles vaccination in atopic dermatitis. PMID- 9357216 TI - No seroconversion after hepatitis B immunization. PMID- 9357217 TI - National Family Welfare Programme: too little, too late. PMID- 9357218 TI - Beta hemolytic streptococci in sore throat infections. PMID- 9357219 TI - Folic acid prevents neural tube defects in high prevalence area. PMID- 9357220 TI - Pregnancy termination for growth retardation--is it useful? PMID- 9357222 TI - Dental practice: a paradigm for ethical change. PMID- 9357221 TI - High-dose dexamethasone for chronic/refractory immune thrombocytopenia. PMID- 9357223 TI - Dental education--a clinical analysis and opinion. PMID- 9357224 TI - Bioethics and Canadian dentistry. AB - Formal ethical reflection and analysis have become expected components of clinical decision making for all health professionals. The implicit acceptance of professional ethics as the sole determinant of "the right and the good" has been replaced by an expectation for more broadly-based understanding of ethical issues. Dentists, like physicians, must be clear about professional ethics and competent in interdisciplinary and interprofessional discussions of the right and good. Modern bioethics is providing approaches to these issues to help practitioners make ethical clinical decisions. These approaches may not be appropriate for dentistry, especially as practised in Canada. Clearly, there are fundamental questions concerning dentistry as a profession, patient-dentist interactions and dental health that must be addressed to form a basis for an ethic relevant to professional practice. The answers to these questions have profound implications for the initial and continuing education of Canadian dentists and for dentistry itself. Some possible starting points for a truly Canadian ethic for dentistry are suggested from a non-dentist, physician ethicist. PMID- 9357225 TI - Legal aspects of dentist-to-dentist referrals. PMID- 9357226 TI - Cotton/resin "wedges" and provisional wire splints. PMID- 9357228 TI - Scientific innovation of 'hysteresis loop' article lauded. PMID- 9357227 TI - Lack of osteopathic component in anxiety supplement. PMID- 9357229 TI - 'Don't eat that hamburger!': examining foodborne illnesses. PMID- 9357230 TI - New medical therapies in glaucoma management. AB - New and potent medications such as alpha 2-adrenergic agonists, topically active carbonic anhydrase inhibitors, and prostaglandin analogs, are now available for use in the treatment of glaucoma. These ophthalmic medications have the potential to cause serious drug interactions and systemic side effects. The primary care physician should be able to recognize and to monitor any potential adverse effects and interact with the prescribing ophthalmologist for optimal patient care. PMID- 9357231 TI - Adverse reactions to fluconazole: illustrative case with focus on desensitization. AB - Fluconazole is an azole antifungal agent. Because it can be taken orally, it is preferred over amphotericin B for long-term treatment. Indications for fluconazole are increasing. Reports of hypersensitivity have been described. If adequate alternative therapy is not available, desensitization may be necessary. Desensitization with fluconazole has not been described. The patient described in this report required fluconazole and was successfully desensitized after manifesting a type 1 hypersensitivity reaction. The patient underwent desensitization during a 15-day period. The starting dose was less than 0.001 dose, with doubling each day. During desensitization, the patient experienced a slight transient rash that resolved with continued therapy. After desensitization, he has continued using fluconazole daily without adverse effects. This report indicates that desensitization to fluconazole can be accomplished safely in selected patients. A suggested desensitization protocol is provided. PMID- 9357232 TI - Comparison of AUDIT and CAGE questionnaires in screening for alcohol use disorders in elderly primary care outpatients. AB - Alcohol use disorders (AUD) can be destructive in the elderly because of drug interactions, higher blood alcohol levels per amount consumed, and limited functional reserve. However, physicians diagnose only about 30% of elderly with AUD. The objective of this study was to screen for AUD in rural elderly family medicine outpatients using the Alcohol Use Disorders Identification Test (AUDIT). A survey of all presenting patients aged 65 years or older who consented (N = 93) was done in four family practices in Southeast Ohio. Measurements included the CAGE and AUDIT questionnaires. On the AUDIT, 13 subjects (14.0%), [10 men, 3 women], screened positive for AUD, scoring 5 or more points, and seven subjects (7.5%) [six men, one woman] screened positive for AUD, scoring 8 or more points. On the CAGE, five men (5.4%) but no women screened positive (> or = 2 affirmatives). The prevalence of AUD found in this survey (5.4% to 14.0%) is consistent with previous studies. Based on these findings, the AUDIT may be a useful screening instrument in the elderly population. PMID- 9357233 TI - Protozoan parasites of the intestinal tract: a review of Coccidia and Microsporida. AB - Ubiquitous in nature, members of the Coccidia and Microsporida are being reported with increasing frequency in the immunocompromised as well as the immunocompetent population. These protozoans are primarily waterborne, but foodborne disease has also been reported. These organisms are responsible for acute, as well as protracted, cases of watery diarrhea with various other related sequelae. The Coccidia includes three genera--Cryptospridium, Isospora, and Cyclospora. The latter two are of lesser importance in terms of morbidity and mortality. The Microsporida includes genera (Enterocytozoon, Encephalitozoon) only recently recognized as important agents of disease. Unlike the Coccidia, these organisms are more restricted to the immunocompromised population. Increased incidence and numbers of patients with prolonged diarrhea due to these forms indicate the need for increased clinical vigilance with regard to prevention, diagnosis, and treatment. PMID- 9357234 TI - Anatomy of an OPTI: Part I. Form, function, and relationships. AB - In July 1995, the American Osteopathic Association (AOA) Board of Trustees passed new regulations for the accreditation of osteopathic graduate medical education (GME) programs by establishing the Osteopathic Postdoctoral Training Institutions (OPTI) system, to be implemented over 4 years. The resulting changes include requirements for college cosponsorship of GME programs and the establishment of standards for the minimum number of residency programs, interns, and residents. The OPTIs will be subject to AOA inspections at least every 5 years. Proponents of the OPTI system claim it will strengthen the profession by promoting educational collaboration, raising academic standards, and requiring appropriate resources to support osteopathic medical education. Opponents fear that it will be too resource intensive, create an additional layer of unnecessary bureaucracy, and have a negative impact on small colleges, hospitals, and states. Despite the controversy, a process for applying for OPTI status has been developed by the AOA, and a number of hospitals and colleges are already developing OPTIs. This article, the first in a two-part series, identifies issues and barriers to be considered in the formation of OPTIs and articulates principles underlying successful collaborations. In Part 2 these issues, principles, and barriers will be reinforced by describing the process used to form a large OPTI--the Ohio University College of Osteopathic Medicine (OU-COM) Centers for Osteopathic Regional Education (CORE) System. PMID- 9357235 TI - Aortopulmonary fistula in an infected Dacron graft for coarctation of the aorta: an uncommon cause of hemoptysis. AB - Hemoptysis secondary to an aortobronchial fistula is rare and uniformly fatal when left untreated. The authors describe a case of massive hemoptysis caused by an aortopulmonary fistula in an infected Dacron graft used successfully to repair a coarctation of the aorta. PMID- 9357236 TI - Magnetic resonance imaging of Takayasu's aortitis in an infant. AB - A fusiform aneurysm involving the ascending aorta and aortic arch in an 8-month old infant was imaged with magnetic resonance. Histologic studies of the excised aneurysm indicated Takayasu's arteritis. Takayasu's arteritis has rarely been reported in infants, and involvement of the ascending aorta and aortic arch is an unusual finding in this age group. PMID- 9357237 TI - Unchecked perinatal mortality in India--problems and challenge. PMID- 9357238 TI - Reflux oesophagitis in acid peptic disease (a fibre-optic endoscopic study). AB - This study is based on entry criteria of visual findings of the upper gastro intestinal fibre-optic endoscopy performed on 100 patients suffering from acid peptic disease (dyspepsia). The diagnostic criteria and grading of reflux oesophagitis advocated by Savary and Miller have been adopted. Reflux oesophagitis was found in 43 cases having grade I oesophagitis in 48.84%, grade II in 32.56%, grade III in 14.00% and grade IV in 4.60% cases. Reflux oesophagitis was associated with hiatus hernia in 23.26%, chronic gastritis in 9.30% of cases. The age varied from 16-80 years with almost equal incidence of male and female, and the severity of the disease increased with the advancement of age. Endoscopy is advised in every case of dyspepsia before any treatment is initiated. PMID- 9357239 TI - Epilepsy in rural Haryana--prevalence and treatment seeking behaviour. AB - A community-based epidemiological study on epilepsy was conducted in rural North India in 1992-94 to estimate the extent of the problem and to study the treatment seeking behaviour of epilepsy patients. A key informant-based survey was conducted by a social worker to enlist suspect epilepsy cases in catchment population (30,000) spreading over in 40 villages of a primary health centre. These cases were verified by a neurologist. The prevalence of epilepsy was 4.2/1000 and it was more in males and in young children (> 6 to 15 years). Majority (81%) had already been diagnosed by qualified doctors. Many (57%) had a fit in last year. In 68% cases the disease onset was in childhood (up to 14 years). Half of the cases had the disease for 6 years or more. Multiple agencies were consulted by most (74%) of them. Faith healers and local practitioners (not medical graduates) were consulted as the first agency in 57% cases. Positive family history was elicited in 36% cases. PMID- 9357240 TI - Profile of electrocardiographic changes in Duchenne muscular dystrophy. AB - Cardiac changes often culminating in cardiac failure are at times a dramatic cause of death in patients of Duchenne muscular dystrophy. These changes are probably invariable in such cases though they may escape detection in early stages by clinical examination or radiological investigation. Electrocardiography serves as a sensitive, non-invasive and inexpensive tool to detect these changes. Fifty-four cases of Duchenne muscular dystrophy were studied clinically and were confirmed by biochemical and electroneuromyographic studies. They were then subjected to electrocardiographic studies. A number of electrocardiographic changes were observed, like tachycardia in 77.77%, deep Q in leads I, aVL, V6 in 53.70%, prolonged VAT in 37.03%, deep Q in leads II, III, aVF in 29.62% and prolonged Q-Tc interval in 25.92% cases. Some of these were distinctive enough to warrant attention for cases of Duchenne muscular dystrophy. PMID- 9357242 TI - Academic performance of various categories of students admitted to Lady Hardinge Medical College, New Delhi. AB - A retrospective study of 4 consecutive academic batches of students was conducted to analyse the performance level of students in 3 university examinations of MBBS. The students were divided into 3 categories based on the criteria of entrance into the medical college. The category I included students who entered through a tough pre-medical test, category II comprised those nominated by government and defence personnel and category III included the Schedule Caste and Schedule Tribe students. The results of the category I was remarkably better than that of II while that of category III was dismal. The last two categories had a large number of students leaving the college without completing their course resulting in 5.4% wastage of precious medical seats and psychological distress. Necessary remedial measures must be planned at the earliest. PMID- 9357241 TI - Role of low dose aspirin in prevention of pregnancy induced hypertension. AB - In a prospective randomised clinical study 50 primi or multigravidae with history of essential hypertension or pregnancy induced hypertension (PIH) in previous pregnancy were selected from antenatal clinics, on the basis of positive roll over test (ROT) carried out between 28 to 30 weeks of pregnancy. The study group comprised 25 women who received 50 mg aspirin daily from day of positive ROT till 37 weeks of pregnancy. Other 25 women served as control. The incidence of PIH in study and control group was 4% versus 28% and that of pre-term birth was 4% versus 24%. The mean birth weight of newborns in the two groups was 3.04 +/- 0.38 kg and 2.71 +/- 0.48 kg respectively. All these differences were statistically significant. No adverse maternal or neonatal complication due to aspirin was observed. PMID- 9357243 TI - Management of macrovascular disease in diabetes mellitus. PMID- 9357244 TI - Argyrophilic nucleolar organiser regions--a new concept in the determination of cytomalignancy. PMID- 9357245 TI - Primary pure red cell aplasia. PMID- 9357247 TI - Lipoleiomyomas of the uterus--a report of 2 cases. PMID- 9357246 TI - Kerosene dependence: a case report. PMID- 9357249 TI - The Consumer Protection Act. PMID- 9357250 TI - The perforation of tuberculous lesion of the intestine is extremely rare. PMID- 9357248 TI - Squamous cell carcinoma arising on the sinus of chronic osteomyelitis of tibia. PMID- 9357251 TI - Cervical cancer. PMID- 9357252 TI - Additives in medicine. PMID- 9357254 TI - Concerning medical treatment of smokers. PMID- 9357253 TI - Consent form for operations. PMID- 9357255 TI - Adolescent pregnancy: a high risk group. AB - In a retrospective study the obstetric behaviour and outcome in 80 teenage pregnancies (< or = 19 years of age) were compared to a control group (n = 80) of women (20-30 years) of same parity. There were 32 booked cases (40%) in study group and 45 (56.2%) in control group (p < 0.05). The overall teenage pregnancy was 3.2%. Majority of patients were 18 years (27.5%) and 19 years (65.0%) in study group and most of them (87.5%) were primiparas. Of all cases 27.5% were grouped under Kuppuswamy classification III in assessing socio-economic status. Anaemia (27.5%), intra-uterine growth retardation (27.5%) and hypertension (15%) were mostly found as complications in study group as compared to controls (11.2%, 8.7% and 8.7% respectively). The incidence of forceps delivery was higher (17.4%) in the study group as compared to controls (6.2%). Stillbirth rate was 1.25% and there was no maternal mortality. PMID- 9357256 TI - Comparative evaluation of fine needle aspiration cytology and biopsy of testis in diagnosis of male infertility. AB - Infertility has a fair degree of male factor contribution in its aetiology, hence needs complete evaluation of male partner especially the status of spermatogenesis. In the present study comparative evaluation of fine needle aspiration cytology (FNAC) and biopsy of testis showed 90% accuracy of FNAC in respect of histopathological diagnosis of spermatogenesis. FNAC is a safe, fairly accurate, outdoor investigation in infertile man and it is devoid of the complications of haematoma formation, suppression of spermatogenesis and antigenic stimulation as seen with testicular biopsy. PMID- 9357257 TI - Pattern of cervical dilatation in previous lower segment caesarean section patients. AB - The pattern of cervical dilatation during labour in 100 patients with previous lower segment caesarean section (LSCS) was determined in a prospective partographic study. Eighty-four subjects delivered successfully by vaginal route. The mean initial dilatation rate (IDR) and average dilatation rate (ADR) were 0.884 cm/hour and 1.255 cm/hour respectively. The mean IDR and ADR of the patients who delivered vaginally were 0.96 cm/hour and 1.41 cm/hour respectively, while of those who required repeat LSCS mean IDR was 0.44 cm/hour and mean ADR was 0.42 cm/hour. Hence ADR in cases who required repeat LSCS was significantly slower as compared to those who delivered vaginally (p < 0.01). Most (87.5%) of the cases who required repeat LSCS crossed the alert line as compared to 34.5% of patients who delivered vaginally. The mean admission delivery interval (ADI) was 9.45 +/- 4.29 hours in patients with no previous vaginal delivery and 8.02 +/- 4.83 hours in patients with previous vaginal delivery. The mean durations of 1st and 2nd stages of labour were 11.8 +/- 5.35 hours and 29.4 +/- 27.3 minutes respectively. It is concluded that partographic evaluation is an important aspect in management of labour of such patients. PMID- 9357258 TI - Congenital abnormalities in Durgapur Steel Plant Hospital with special reference to neural tube defect. AB - In a retrospective study of congenital abnormalities from January, 1991 to December, 1993 in Durgapur Steel Plant Hospital, a referral hospital covering wide industrial belt, the incidence of neural tube defects was detected remarkably high. It was revealed that increased incidence of neural tube defects was found in patients from low socio-economic status with gross nutritional deficiency and inadequate antenatal care. PMID- 9357259 TI - Practice of standards in female sterilisation. AB - During training of trainers (TOT) courses organised for medical personnel of Haryana Civil Medical Services (HCMS) by COE Medical College, Rohtak, 55 doctors involved in female sterilisations were interrogated regarding practices in counselling, informed decision, asepsis, surgical procedure, operative and postoperative care and follow-up of the clients accepting sterilisation as contraception. Counselling was the responsibility of auxiliary nurse cum midwife (ANM) lady health volunteer (LHV)/other paramedical workers as viewed by 89.1% participants whereas 85.4% thought that the registration clerk should take the informed consent. Eligibility criteria were always adhered to by 10.9% participants. Asepsis and sterilisation of instruments, etc, were maintained by operation theatre (OT) attendant or OT nurse as answered by 90.9% doctors. Skin preparation was done by a solution containing cetrimide and chlorhexidine alone by 70.8% doctors. The ligation and excision was the method practised by all. Catgut suture was used by only 43.6% doctors. Twenty-six maternal deaths were reported by 20 participants during their whole career. There were 7 deaths on the table, all with laparoscopic sterilisation. Peritonitis with septicaemia was the major cause of death in majority of cases. To ensure high quality and safety of voluntary surgical contraception, programmes must establish a system to ensure that standards are maintained. PMID- 9357260 TI - Analgesic nephropathy. PMID- 9357261 TI - Finance Bill, 1997--important income-tax amendments. PMID- 9357262 TI - Reifenstein syndrome: report of 3 cases. PMID- 9357263 TI - Hypoplastic left heart syndrome: a report of two cases with review of literature. PMID- 9357264 TI - Lymphangioma of the ovary. PMID- 9357266 TI - Risk factor hypothesis. PMID- 9357265 TI - Intra-ocular malignant medullo-epithelioma--two cases reported in ten years. PMID- 9357267 TI - Quo vadis, IMA. PMID- 9357268 TI - Free radicals and aging. PMID- 9357269 TI - Pedicled omental transfer for ischaemic limbs--a 5-year experience. AB - Chronic occlusive arterial diseases form a single largest entity amongst the peripheral vascular diseases. Current operative methods available for improving circulation often elicit poor results and the patient has to undergo an amputation. The technique of pedicled omental transfer has given hope of saving such unsalvageable limbs. Although symptomatic and clinical improvement has been reported by this method of "biological by-pass revascularisation", there are no simple, objective and easily reproducible tests to assess improvement in circulation. In this study pulse oximetry and stress testing have been used to assess revascularisation. This study comprised 56 patients (78 limbs) suffering from chronic occlusive arterial disease, spanning a period of 5 years. Patients were investigated and subjected to pedicled omental transplantation (omentopexy). Symptomatological assessment showed improvement in intermittent claudication in about 85% of patients, relief from rest pain in 86% and healing of chronic ulcers in 73% of patients. Objective tests of stress testing and pulse oximetry also showed improvement in circulation. Relief from ischaemia was more in cases of Buerger's disease (TAO) than in cases of atherosclerosis obliterans (ASO). PMID- 9357270 TI - Impact of mode of delivery on maternal mortality in eclampsia. AB - Determinants of maternal mortality and causes of death pertaining to mode of delivery have been discussed. There were 23 deaths (case fatality rate of 7.2%) and maximum deaths occurred in intrapartum eclampsia (12 ie, 52.17%). Caesarean section was performed in 92 cases (28.7%) of which 4 women died (4.3%). Maternal mortality in cases who delivered vaginally was 7.1% (16 out of 225) and 3 cases died undelivered. Authors feel that at the referral centres early caesarean section in eclampsia may help in reducing maternal mortality. PMID- 9357272 TI - Pattern of solid malignant tumours in children--a ten-year study. AB - Solid malignant tumours (n = 263) excluding brain and spinal cord tumours in children up to 14 years of age were studied. Retinoblastoma (27%) constituted the largest group followed by Wilms' tumour (14.1%) and lymphoma (13.7%). Most patients (55%) were of less than 5 years age and maximum incidence of embryonal tumours was found in this age group; other tumours were more frequent in higher age. A male preponderance was noted (male to female ratio as 1.6:1). Amongst lymphoma, 61% were non-Hodgkin's lymphoma and rest were Hodgkin's disease; 2 cases of Burkitt's lymphoma were found. Other notable tumours encountered in the study were embryonal rhabdomyosarcoma (n = 14), hepatoblastoma (n = 9), neuroblastoma (n = 7), Ewing's sarcoma (n = 21), osteogenic sarcoma (n = 19) and germ cell tumours (n = 14). PMID- 9357271 TI - Role of epidural tramadol hydrochloride on postoperative pain relief in caesarean section delivery. AB - Two groups comprising 25 patients in each went for caesarean section delivery under epidural anaesthesia. Group I patients received 50 mg (1 ml) of tramadol hydrochloride with 14 ml of 2% lignocaine with adrenaline (1:200,000) and group II cases received 15 ml of 2% lignocaine with adrenaline (1:200,000). Both the groups of patients were comparable in age and body weight. In both the groups, there were good operative conditions, insignificant changes in pulse and blood pressure. The neonatal status was also similar in both the groups. The patients belonging to group I showed longer duration (15.39 +/- 0.45 hours) of analgesia in comparison to group II patients (2.46 +/- 0.54 hours). PMID- 9357274 TI - If you 'possess' it, you pay for it. PMID- 9357273 TI - Exchange transfusion in neonates. PMID- 9357275 TI - Multiple myeloma: unusual presentation as mono-articular arthritis. PMID- 9357276 TI - Pseudo-crossed renal ectopia. PMID- 9357277 TI - Non-clinical delivery systems: an alternative approach for distribution of iron and folic acid tablets. PMID- 9357278 TI - Correspondence certificate course by the IMACGP. PMID- 9357279 TI - A new epidemic. PMID- 9357280 TI - Carcinoma of the ovary masqurading as gall bladder neoplasm. PMID- 9357281 TI - A case of suburethral growth. PMID- 9357282 TI - Presence of both testes on the same side. PMID- 9357283 TI - Congenital haemangioma of the tongue. PMID- 9357284 TI - Perineal breast. PMID- 9357285 TI - Slipped catheter in urinary bladder. PMID- 9357286 TI - Epidermoid cyst of brain--an incidental autopsy finding. PMID- 9357288 TI - Surgical management of intrathoracic goiter. AB - Intrathoracic goiter is a rare clinical entity. This goiter can develop slowly and the patient may be asymptomatic for many years. A significant number of these patients, however, may develop various complications as a result of compression of vital structures or malignancy. In this report, the surgical management of 18 cases of intrathoracic goiter are reviewed. Six of the patients underwent right thoracotomy with resection of the posterior mediastinal goiter. The remaining 12 patients had their tumor removed via median sternotomy. There were no surgical deaths within this series. Although the incidence of intrathoracic tumor is limited, the threat of malignancy and compressive symptoms clearly indicates the need for surgical excision. Our group recommends thoracotomy and median sternotomy for surgical excision. PMID- 9357287 TI - Accessory paragonadal spleen simulating testicular neoplasm. PMID- 9357289 TI - 25 years--a look back. PMID- 9357290 TI - Computer requirements for medical school students--implications for admissions. AB - Computers are increasingly being used in clinical practice settings. Aware of the need to educate students regarding computer applications in medicine, the University of Kentucky College of Medicine is in the midst of developing a computer curriculum. To that end, many courses and clerkships have devised software packages for transmittal of course information and for evaluation of student performance. This paper outlines requisite computer skills that applicants applying to medical school should possess, broadly reviews how those computer skills will be used in medical school, and suggests means for attaining computer competency prior to making application to medical school. PMID- 9357291 TI - Determinability of model parameters in a two-stage deterministic cancer model. AB - We investigate the determinability of model parameters for a two-stage cancer model, using as an example chemically induced skin cancer in mice. When the time course of the papilloma number and cancer rate are known, the rate of creation of initiated cells and their net-growth rate can be uniquely determined. These two high-level parameters are sufficient to uniquely simulate the experimental papilloma data. However, the mitotic and death rates of initiated and transformed cells cannot be determined from the available experimental data. The rate of creation of transformed cells and their net-growth rate cannot be determined independently. Thus, although the deterministic two-stage cancer model can simulate the kinetics of papilloma formation and skin cancer data, many of the basic underlying biological parameters (i.e., mitotic and death rates) cannot be uniquely determined from the usually available biological data. PMID- 9357292 TI - Backward bifurcation in epidemic control. AB - For a class of epidemiological SIRS models that include public health policies, the stability at the uninfected state and the prevalence at the infected state are investigated. Backward bifurcation from the uninfected state and hysteresis effects are shown to occur for some range of parameters. In such cases, the reproduction number does not describe the necessary elimination effort; rather the effort is described by the value of the critical parameter at the turning point. An explicit expression is given for this quantity. The phenomenon of subcritical bifurcation in epidemic modeling is also discussed in terms of group models, pair formation, and macroparasite infection. PMID- 9357293 TI - Dynamic consequences of reproductive delay in Leslie matrix models with nonlinear survival probabilities. AB - The dynamic consequences of reproductive delay in Leslie matrix models with nonlinear survival probabilities p are analyzed. In consideration of two-age classes, proof is presented for a wide range of p functions that, outside the strongly resonant cases, the transfer from stability to instability goes through a supercritical Hopf bifurcation and, moreover, that the nonlinear development has a strong resemblance of three or four cycles, either exact or approximate. In three-age class models, the tendency toward four-periodical dynamics is shown to be even more pronounced, a qualitative finding that gradually disappears as we turn to the higher-dimensional cases. We also prove that for models of any dimension n > 1 theme are regions in parameter space where the equilibrium is unstable at its creation and we demonstrate that the dynamics in this age-class extinguishing case is 2k.n cyclic. PMID- 9357294 TI - "Strange young women on errands". Obstetric nursing between two worlds. PMID- 9357296 TI - "For Zion's sake": the emergence of Mormon nursing. PMID- 9357298 TI - Called to a mission of charity. The Sisters of St. Joseph in the Civil War. PMID- 9357295 TI - Political women, professional nurses, and the creation of Alberta's District Nursing Service, 1919-1925. PMID- 9357297 TI - An experiment in leadership. The rise of student government at Philadelphia General Hospital Training School, 1920-1930. PMID- 9357299 TI - "A somewhat duskier skin": Mary Seacole in the Crimea. PMID- 9357300 TI - Alternative visions. The nurse-technology relationship in the context of the history of technology. PMID- 9357301 TI - Synergistic sterilization. PMID- 9357302 TI - Stability of deslorelin injection. AB - The stability of deslorelin, a nonapeptide drug, in an injectable drug formulation was studied for 60 months. An HPLC analytical method was developed to quantify deslorelin and benzyl alcohol in deslorelin injection. The samples containing 500 micrograms/mL of deslorelin were stored at--10, 6, 25, 40 and 50 degrees C. No deslorelin degradation was observed in samples stored at 6 degrees C, but the loss of deslorelin in samples stored at 25, 40 and 50 degrees C was found to increase with temperature. The T90 of the product at 6 degrees C was estimated to be greater than 20 years from the short-term stability data of four lots stored at 25, 40 and 50 degrees C. The long-term stability results confirmed that there was no degradation of deslorelin stored at 6 degrees C for 5 years. Liquid secondary ionization mass spectrometry analysis of aged samples showed hydrolysis of deslorelin and the production of a hexapeptide (< Ser-Tyr-D-Trp-Leu Arg-Pro-NH-Et) with a molecular weight of 847. A second product resulted from the oxidation of the tryptophan indoylyl ring. The oxyindoyle decapeptide structure had an ion m/z of 1298. The mass spectrum of pure deslorelin also is reported. PMID- 9357303 TI - Preformulation studies for the development of a parenteral liquid formulation of an antitumor agent, AG337. AB - AG337 is a potential anticancer agent designed by using protein structure-based techniques. The objective of this work was to evaluate the feasibility of a high concentration liquid formulation of AG337 intended for intravenous administration. The solubility of AG337 in pure water was above 100 mg/mL at pH < 3. The drug's solubility decreased precipitously as the solution pH increased above 3 upon titration with 0.1 N NaOH. The solubility of AG337 in water as a function of temperature (ranged from 2-40 degrees C) was determined. As anticipated, the drug's solubility increased somewhat linearly as the solution temperature increased. Degradation kinetics of 15% and 10% AG337 solutions at elevated temperatures was determined to assess the feasibility of a liquid formulation as opposed to previously developed lyophilized powder for injection. Only one major degradation product was detected in the HPLC as a result of chemical hydrolysis of AG337 to AG408. Arrhenius plot (i.e., kobs versus 1/T) revealed an activation energy of 25 kcal/mol. The shelf life (t95%) of 10% AG337 solution of pH 2 at 25 degrees C was predicted to be roughly 8 years. Various terminal sterilization methods, which include moist/dry autoclaving (121 degrees C), e-beam, and gamma-irradiation, were evaluated for the 10% AG337 solution. Autoclaving cycles, ranged from 20 to 90 minutes, caused instantaneous degradation of AG337 solution and induced further degradation upon long-term storage. Again, AG408 was the major degradation product following autoclaving. On the other hand, irradiation techniques induced very little degradation, but turned clear glass vials to brown upon irradiation. PMID- 9357304 TI - Pharmaceutical container/closure integrity. I: Mass spectrometry-based helium leak rate detection for rubber-stoppered glass vials. AB - The development of mass spectrometry-based leak detection for pharmaceutical container integrity was undertaken to provide an alternative to microbial challenge testing. Standard 10-mL vials were modified to contain pinholes (0.5 to 10 microns) by affixing micropipettes with epoxy into 2-mm vial side wall holes. The absolute leak rate was determined using vials that were sealed in a tracer (helium) environment with butyl rubber stoppers and crimps. Alternatively leak rates were determined using vials that were sealed in room air and exposed to tracer under pressure (charging or bombing). Tracer leak rates were measured with mass spectrometry leak rate detectors. The absolute leak rate was correlated the squared nominal leak radius which suggested that the mode of gas flow through the glass pipette leaks was more turbulent than viscous even at low leak rates typically associated with viscous flow. The minimum observed absolute leak rate was about 10(-6.6) std cc/sec and was likely due to helium permeation through the rubber stoppers. Heat-stressed rubber stoppers did not affect the baseline absolute leak rate. Adsorption of helium tracer to the test unit surfaces was found to confound baseline leak rate measurement reliability but was eliminated as a source of variation by exposing the test units to ambient air for > or = 12 hours. The absolute leak rate and the leak rate measured after charging were related in a mathematically predictable way. PMID- 9357305 TI - Pharmaceutical container/closure integrity. II: The relationship between microbial ingress and helium leak rates in rubber-stoppered glass vials. AB - Helium leak rate measurements were quantitatively correlated to the probability of microbial ingress for rubber-stoppered glass vials subjected to immersion challenge. Standard 10-mL tubing glass vials were modified by inserting micropipettes of various sizes (0.1 to 10 microns nominal diameter) into a side wall hole and securing them with epoxy. Butyl rubber closures and aluminum crimps were used to seal the vials. The test units were sealed in a helium-filled glove bag, then the absolute helium leak rates were determined. The test units were disassembled, filled with media, resealed, and autoclaved. The test units were thermally treated to eliminate airlocks within the micropipette lumen and establish a liquid path between microbial challenge media and the test units' contents. Microbial challenge was performed by immersing the test units in a 35 degrees C bath containing magnesium ion and 8 to 10 logs of viable P. diminuta and E. coli for 24 hours. The test units were then incubated at 35 degrees C for an additional 13 days. Microbial ingress was detected by turbidity and plating on blood agar. The elimination of airlocks was confirmed by the presence of magnesium ions in the vial contents by atomic absorption spectrometry. A total of 288 vials were subjected to microbial challenge testing. Those test units whose contents failed to show detectable magnesium ions were eliminated from further analysis. At large leak rates, the probability of microbial ingress approached 100% and at very low leak rates microbial ingress rates were 0%. A dramatic increase in microbial failure occurred in the leak rate region 10(-4.5) to 10(-3) std cc/sec, which roughly corresponded to leak diameters ranging from 0.4 to 2 microns. Below a leak rate of 10(-4.5) std cc/sec the microbial failure rate was < 10%. The critical leak rate in our studies, i.e. the value below which microbial ingress cannot occur because the leak is too small, was observed to be between 10(-5) and 10(-5.8) std cc/sec, which corresponds to an approximate leak diameter of 0.2-0.3 micron. PMID- 9357306 TI - Pharmaceutical container/closure integrity. III: Validation of the helium leak rate method for rigid pharmaceutical containers. AB - Validation of a helium leak rate method for pharmaceutical container/closure integrity quality assurance required the demonstration that this physical testing method was as good or better than microbial immersion challenge testing in detecting potential integrity failures. One lot of rubber-stoppered, broth-filled glass vials also containing defective vials with known leaks were subjected to both helium leak rate and microbial challenge testing. The defective vials were prepared by affixing glass micropipettes (0.1 to 10 microns) into the vial side walls. The validation lot included a 10% seeded defect rate of which about 50% contained leaks with a predicted probability of failing a microbial challenge (> 10%). Helium tracer was placed in the test units by charging them for 4 hours under a 40 psi helium pressure. The critical leak rate after charging was determined to be 10(-7) standard cc/second, and test units with measured leak rates greater than this value were considered helium leak rate failures. Microbial immersion challenge was conducted by exposing the test units in a bath inoculated with 10(9-10) viable E. coli and B. diminuta organisms for 24 hours followed by a 13 day (35 degrees C) incubation. Microbial failures were determined visually. The helium and microbial leak test methods were compared statistically using mean failure rates. The mean helium failure rate was 6.9%, whereas the mean microbial failure rate was 2.8%. The difference between helium and microbial failure rates was significantly greater than zero. Thus, helium leak rate testing was demonstrated to be a suitable pharmaceutical container/closure integrity method for microbial quality assurance of rigid containers. PMID- 9357307 TI - Cumulative viral titer reduction demonstrated by sequential challenge of a tangential flow membrane filtration system and a direct flow pleated filter cartridge. AB - The efficiency of sequential incorporation of two approaches to virus removal, i.e., tangential flow ultrafiltration using the Omega 300K VR Maximate System, and, direct flow microfiltration using the Ultipor VF grade DV50 virus removal filter, was evaluated using bacteriophage phi 6 in Dulbecco's modified Eagle medium (MEM) as the carrier fluid. A combined log titer reduction value of > 10 was demonstrated, with each step, individually, providing > 4.5 log reduction. Significant improvement in filter life was also demonstrated when the combined system was used. The data suggest that this system may have applications in the manufacture of biologicals and biopharmaceuticals, where there is a requirement for multiple robust methods of virus removal to ensure freedom from endogenous or adventitious viral contamination. PMID- 9357308 TI - PDA comments to USP on elastomeric closures for injection (381). Parenteral Drug Association. PMID- 9357309 TI - Extinction is forever. PMID- 9357310 TI - MDM2--arbiter of p53's destruction. PMID- 9357311 TI - SUMO-1: wrestling with a new ubiquitin-related modifier. PMID- 9357312 TI - Endosomal proteases and antigen processing. AB - T cells are activated by fragments of antigenic proteins bound to major histocompatibility complex (MHC) molecules and displayed on the cell surface. MHC class II proteins scavenge processed protein antigens from within endosomal compartments. The antigenic peptides are generated within these and other intracellular compartments using the array of proteolytic enzymes normally involved in terminal protein degradation. Antigen-presenting cells use different mechanisms to exploit and control the activity of these enzymes so as to ensure the generation of a wide variety of peptides, while preventing the destruction of antigenic epitopes by excessive proteolysis. PMID- 9357313 TI - The ubiquitin system. AB - Eukaryotes contain a highly conserved multi-enzyme system that covalently links ubiquitin to a variety of intracellular proteins that bear degradation signals recognized by this system. The resulting ubiquitin-protein conjugates are degraded by the 26S proteasome, a large ATP-dependent protease. Pathways that involve ubiquitin underlie a multitude of processes, including cell differentiation, the cell cycle and responses to stress. PMID- 9357314 TI - Caspases and caspase inhibitors. AB - Five years ago, little was known about mechanisms of apoptotic execution. Now, one class of cell-death gene, the cysteine and aspartases (caspases) has come under intensive study. This review discusses the two classes of caspases, the reasons why humans may have so many caspase genes, the growing list of caspase substrates, and viral and pharmacological caspase inhibitors. PMID- 9357315 TI - Temporal and spatial control of the adenovirus proteinase by both a peptide and the viral DNA. AB - The adenovirus proteinase (AVP) uses both an 11-amino acid peptide (pVIc) and the viral DNA as cofactors to increase its catalytic rate constant 6000-fold. The crystal structure of an AVP-pVIc complex at 2.6-A resolution reveals a new protein fold of an enzyme that is the first member of a new class of cysteine proteinases, which arose via convergent evolution. PMID- 9357317 TI - The tortuous story of Asp ... His ... Ser: structural analysis of alpha chymotrypsin. PMID- 9357316 TI - Self-compartmentalizing proteases. AB - Among the hundreds of proteases characterized so far, most of which are monomeric or dimeric, there is a small group that form compartments through self association and that segregate their proteolytic active sites to the interior of these compartments. Although few in number, they represent the main agents of intracellular protein breakdown. They belong to different hydrolase families but have converged towards the same barrel-shaped architecture. Frequently, they are coupled to chaperone-like ATPases of similar quaternary structure that regulate the access to the proteolytic compartments and appear to have been recruited from the same branch of P-loop NTPases. PMID- 9357318 TI - Potassium channels still hot. PMID- 9357319 TI - The dual nature of the tachykinin NK1 receptor. PMID- 9357320 TI - A three-state receptor model of agonist action. AB - The concept that receptors can exist in multiple conformational states is becoming a physical reality. A fundamental question is how many active states need to be proposed in order to account for pharmacological observations, in particular, the finding that the same receptor type can exhibit a different agonist pharmacology when coupled to different effector pathways. In this article, Paul Leff, Clare Scaramellini, Clare Law and Ken McKechnie propose and develop a three-state receptor model in which two active conformations are assumed to exist. They show that this relatively simple theoretical model provides a basis for predicting variable agonist and inverse agonist behaviour in different systems containing the same receptor, and that it is able to account for emerging data obtained on promiscuously coupled receptors. It is argued that, while these new theoretical considerations challenge the fundamental assumptions and concepts of traditional receptor theory, the general principles of pharmacological receptor classification are largely preserved. PMID- 9357321 TI - Identifying neuronal non-L Ca2+ channels--more than stamp collecting? AB - The pharmacology of the majority of Ca2+ channels in the nervous system is very limited. Although attempts have been made to constrain native Ca2+ channels into the framework provided by the six pore-forming molecules cloned to date, refined biophysical analysis of Ca2+ currents, expression techniques and the use of selective toxins have helped to identify unambiguously only a limited number of Ca2+ channels. In fact, many native Ca2+ channel activities remain as 'orphans', waiting for their molecular counterparts to be defined. In this article, Janet Nooney, Regis Lambert and Anne Feltz systematically delineate the well characterized non-L Ca2+ channel activities and the missing elements in our knowledge of the Ca2+ channel family. PMID- 9357322 TI - Nonpeptide antagonists of neuropeptide receptors: tools for research and therapy. AB - The recent development of selective and highly potent nonpeptide antagonists for peptide receptors has constituted a major breakthrough in the field of neuropeptide research. Following the discovery of the first nonpeptide antagonists for peptide receptors ten years ago, numerous other antagonists have been developed for most neuropeptide families. These new, metabolically stable compounds, orally active and capable of crossing the blood-brain barrier, offer clear advantages over the previously available peptide antagonists. Nonpeptide antagonists have provided valuable tools to investigate peptide receptors at the molecular, pharmacological and anatomical levels, and have considerably advanced our understanding of the pathophysiological roles of peptides in the CNS and periphery. Evidence from animal and clinical studies suggests that nonpeptide antagonists binding to peptide receptors could be useful for the treatment of disease states associated with high levels of neuropeptides. In this article Catalina Batancur, Mounia Azzi and William Rostene will address the recent developments in nonpeptide antagonists for neuropeptide receptors, with a particular focus on their CNS actions. PMID- 9357323 TI - Potential signalling roles for UTP and UDP: sources, regulation and release of uracil nucleotides. AB - Increasing evidence for receptors for uracil nucleotides has focused interest on specific signalling mechanisms involving UTP and UDP. At least three metabotropic P2 receptors are stimulated by uracil nucleotides with equal or greater potency than by adenine nucleotides, and there might be ionotropic receptors as well. Regulation of uridine and uracil nucleotide levels is important when considering the receptor-mediated effects of these compounds. Cells can synthesize uracil nucleotides de novo or by salvage of uridine. UTP made from salvage might be preferentially used for RNA synthesis in the nucleus, while UTP synthesized de novo seems to be used for UDP-sugar and CDP-phospholipid production. UTP from both pathways can enter a free UTP pool, from which UTP can be released from cells. UTP and UDP can stimulate pyrimidinoceptors, but metabolism by ecto nucleotidases limits their effects. Alternatively, UTP might be a substrate for ecto-protein kinases, and this could contribute to its extracellular regulation. Cells can reclaim uridine, using nucleoside transport processes, following dephosphorylation of UTP, UDP and UMP. In this article Christopher Anderson and Fiona Parkinson discuss how understanding the processes that regulate uridine and uracil nucleotide concentrations will enhance our ability to manipulate UTP/UDP signalling pathways for pharmacological effect. PMID- 9357324 TI - Interstitial cells of Cajal as targets for pharmacological intervention in gastrointestinal motor disorders. AB - Interstitial cells of Cajal (ICCs) have recently been identified as the pacemaker cells for contractile activity of the gastrointestinal tract. These cells generate the electrical 'slow-wave' activity that determines the characteristic frequency of phasic contractions of the stomach, intestine and colon. Slow waves also determine the direction and velocity of propagation of peristaltic activity, in concert with the enteric nervous system. Characterization of receptors and ion channels in the ICC membrane is under way, and manipulation of slow-wave activity markedly alters movement of contents through the gut organs. Here Jan Huizinga, Lars Thuneberg, Jean-Marie Vanderwinden and Juri Rumessen, suggest that, as ICCs are unique to the gut, they might be ideal targets for pharmacological intervention in gastrointestinal motility disorders, which are very common and costly. PMID- 9357325 TI - Expanded Programme on Immunization (EPI). PMID- 9357326 TI - [Structures and functions of ribonucleases]. AB - 1. In order to understand the differences in pH optima and reaction rates of RNase A towards low molecular weight substrates and polymer substrates, the subsite structure of bovine pancreatic RNase A was studied. The kinetic studies of various sizes of oligouridylic acids showed that the size of the subsite is three nucleotides long. The kinetic studies on the inhibition of pUp, X-ray crystallographies of RNase A-ApC and pTp complexes, 31P-NMR studies on the binding of RNase A-pAp, and pTp showed the presence of P0, P2 and B3 sites. The location of the P0 site was assigned to be Lys66 by X-ray crystallography of the RNase A-pTp complex. The location of the P2 and/or P3/B3 site was determined by studying the enzymatic activities of several S-peptide analogs in which N-Leu was substituted for Lys7 and/or Lys1 coupled with S-protein toward various chain lengths of oligouridylic acids. The experiment suggested that P2 is Lys7 and P3/B3 is Lys1. 2. Several new pyrimidine base specific RNases were isolated and their primary structures were determined. They were two non-secretory RNases, a bovine liver alkaline RNase, a bovine brain RNase, and a bullfrog liver RNase. The bovine brain RNase has extra 16 amino acids at the C-terminus with O glycosylated Ser. The bullfrog liver RNase was an extremely heat-stable RNase so far known. 3. Two new RNases belonging to RNase T1 family were isolated and their primary structures were elucidated. They were RNases isolated from Aspergillus saitoi and a mushroom (hiratake). The former RNase has a similar structure to RNase T1, but it was a base non-specific and guanylic acid preferential enzyme. From the results of X-crystallographic studies of this RNase, we suggested that the mechanism of RNase T1 RNase is essentialy a general acid-base catalysis between His40 and Glu58. 4. We isolated several fungal, plant and animal base non specific acid RNases with a molecular mass about 24 kDa or more, and elucidated their primary structures. These RNases contain two sequences containing common 7 8 amino acid residues in common which include most of the amino acid residues important for the catalysis. Therefore, we proposed to designate these RNases as RNase T2 family RNase. On the basis of chemical modifications, kinetic studies and protein engineering studies of RNase Rh from Rhizopus niveus and RNase M from A. saitoi, we assigned that the catalytic site of RNase Rh consists of His46, His104, His109, Glu105, and Lys108. In the mechanism we proposed for RNase Rh, His46 and His109 work as a general acid and base catalysts. His104 was a phosphate binding site, and Glu105 and Lys108 might work to polarize a P=O bond of the substrate or stabilize the pentacovalent intermediate. However, in the reverse reaction of the transfer reaction step and the hydrolysis step of RNase Rh, His109 and His46 work as an acid and base catalyst, respectively. The X-ray crystallographic studies of RNase Rh, an RNase Rh-2'-AMP or d(ApC)complex, and the protein engineering studies of several mutant enzymes assigned the components of the major base recognition site (B1 site) and the minor base recognition site (B2 sites) of RNase Rh. The enzymatic studies of several mutant enzymes indicated that (i) Asp51 is very crucial for adenine base recognition, and the replacement of Asp51 by other amino acid, such as Thr, Ser, Glu, Asn makes RNase Rh more guanylic acid preferential, (ii) the replacement of Trp49 by Phe, and Tyr57 by Trp make the enzyme more pyrimidine and purine bases preferential, respectively. These trials are the first example of marked artificial change in the base specificity of RNases. PMID- 9357327 TI - [Transformation of natural products into more potent compounds: chemical modification of monensin]. AB - Monensin (1) is a representative compound of polyether ionophore antibiotics, which selectively transport Na+ ions. In order to obtain potent Na+ ionophores, the modification of the carboxyl group of monensin was carried out to yield monensylamino acids (2) and monensylamino acid-1,29-lactones (3). The Na+ permeability of ion through the erythrocyte membrane of 2 and 3 was evaluated by the 23Na-NMR method. Compound 2 showed less Na+ ion transport activity than monensin, probably due to the lower lipophilicity caused by the conformational change of the chain moiety of the molecules. Although 3 showed higher lipophilisity than 1, 3 had no Na+ ion permeability, probably due to loss of the carboxyl group. As more lipophilic compounds possessing a carboxyl group was supposed to have more ion transport activity, 7-O-acylmonensins (8) and 7-O alkylmonensins (11) were synthesized. Among these compounds, the value of Na+ ion permeability of 7-O-benzylmonensin (11c) was 1.4 time that of 1. Further investigation was carried out by preparing various 7-O-(substituted benzyl)monensins (13), and 7-O-(p-ethylbenzyl)monensin (13b) exhibited the largest Na+ ion permeability, about twice the value of 1. In order to convert monensin (1) to Ca2+ ionophore, 7-carboxylmethylmonensin (18) via protected 7 oxomonensin (15), and 25-carboxylmonensin (26) were prepared. In the course of the synthesis, 15 was clarified as a useful intermediate to give 7-amino and 7 alkyl derivatives. Ca2+ ion transport activities of 18 and 26 were determined by a CHCl3 liquid membrane system. 25-carboxylmonensin (26) showed 70% of the activity of Ca2+ ionophore, lasalocid A, and compound 26 could be the lead compound for the preparation of a new Ca2+ ionophore. PMID- 9357328 TI - [Studies on compounds with antioxidant activity--development of hypoglycemic agents, troglitazone (CS-045)]. AB - An interest in compounds having both the quenching activity of the active oxygen species (antioxidant activity) and hypoglycemic and/or hypolipidemic activities significantly increases in the field of angiopathic disease, especially in the elderly. We studied hindered phenols in order to find biologically active compounds having the above activities. As a result, several compounds, such as 1,3-benzoxathioles, phenoxypentanoic acids, phenoxypentanols, 2-chloro-3 phenylpropionic acids, and thiazolidines, were found. Among these compounds, a thiazolidine, troglitazone, (CS-045), 1, showed desired biological properties both in vitro and in vivo without causing any increase in liver weight, and so was developed as a new type of antidiabetic agent. In this review, the structural design and the biological activities including antioxidant activities are described. PMID- 9357329 TI - [Studies on 5-lipoxygenase inhibitors. Synthesis of and structure--activity relationship in p-hydroxyarylalkenylbenzoazoles]. AB - In a previous paper we reported that 2-(p-hydroxyarylbutadienyl)benzoxazoles are highly potent 5-lipoxygenase inhibitors. We synthesized their ethenyl homologues and benzothiazole derivatives, and evaluated their 5-lipoxygenase inhibitory activity in vitro with cell-free rat basophilic leukemia (RBL-1). In most cases the replacement of benzoxazolyl with benzothiazolyl resulted in an enhancement of the activity. All compounds with butadienyl spacers tested herein exhibited strong inhibitory activities. While most of the ethenyl homologues showed weaker activities than their corresponding butadienyl homologues, some ethenyl compounds in the benzothiazole derivatives were found to be as potent as their corresponding butadienyl homologues. The inhibitory activity was also affected by the variation in the p-hydroxyaryl functionality. PMID- 9357330 TI - [Studies on antioxidant effect of Hypsizigus marmoreus. I. Effects of Hypsizigus marmoreus for antioxidant activities of mice plasma]. AB - The antioxidant effects of Hypsizigus marmoreus, one of the most popular Japanese edible mushrooms, were investigated by the use of the augmentation effect of antioxidant activity (AOA) in the mice plasma. An aqueous extract of the mushroom fruit-body was found to have a slight trap activity for peroxyl and alkoxyl radicals. On the other hand, the blood plasma of mice fed with a fodder containing 5-10% of the dried powder of the mushroom extract augmented significantly AOA for alkoxyl radicals. It was suggested from analysis of the plasma by the HPLC post column AOA method that the increase of AOA in the mice plasma was caused by the induction of high molecular weight fractions having AOA produced by feeding Hypsizigus marmoreus. The amount of lipoperoxide in the mice plasma as values of thiobarbituric acid reactive substances showed a tendency to be lowered by intake of Hypsizigus marmoreus. These results suggest that oral administration of the fruit-body of Hypsizigus marmoreus can induce an antioxidant effect in the mice plasma. PMID- 9357331 TI - [Sustained release double-layered progesterone suppository for luteal support therapy]. AB - A sustained release suppository containing progesterone with a double-layered structure was prepared for the treatment of the luteal phase defect. Hydroxypropylcellulose-H (HPC) and Carbopol-934P (CP) were used as bases of the inner layer and Witepsol W35 was used as a base of the outer layer. The strength of the inner layer (stick) decreased with the increase of the rate of content of HPC component. The strength of the stick which was prepared from a mixture of HPC and CP in a ratio of 1:1, was inverse by proportional to the rate of the addition of crystalline cellulose (CC) and the amount of released drug was proportional to the rate of the addition of CC. The area under the drug release curve of the stick containing 60% of CC in the base was about 12 times of the stick containing no CC (control stick). Furthermore, the mean release time of the stick containing 60% of CC became about a half of the control stick. It was suggested that the drug release of progesterone from the stick could be controlled by changing the rate of the addition of CC. Two types of suppository which containing progesterone in both phases (suppository A) and in the stick alone (suppository B) were prepared. Both suppositories showed a sustained release property and suppository B had a lag time of two hours. When the suppositories were administered in to the vagina of rabbits, they showed a sustained release property and a rapid rise in the serum concentration was more suppressed than an ordinary Witepsol suppository. One hour after the administration of the two layered suppository, some parts of the suppository was identified macroscopically to be remained in the vagina. The usefulness of the double-layered suppository as a hospital preparation should be suggested after the attainment of the optimization of the formulation. PMID- 9357333 TI - Grading leniency is a removable contaminant of student ratings. AB - It is well established that students' evaluative ratings of instruction correlate positively with expected course grades. The authors identify 4 additional data patterns that, collectively, discriminate among 5 theories of the grades--ratings correlation. The presence of all 4 of these markers in student ratings data (obtained at University of Washington) was most consistent with the theory that the grades--ratings correlation is due to an unwanted influence of instructors' grading leniency on ratings. This conclusion justifies use of a statistical correction--illustrated here with actual ratings data--to remove the unwanted inflation of ratings produced by lenient grading. Additional research can profitably seek other inappropriate influences on ratings to identify more opportunities for validity-enhancing adjustments. PMID- 9357332 TI - Validity concerns and usefulness of student ratings of instruction. AB - The validity of student rating measures of instructional quality was severely questioned in the 1970s. By the early 1980s, however, most expert opinion viewed student rating measures as valid and as worthy of widespread use. In retrospect, older discriminant-validity concerns were not so much resolved as they were displaced from research attention by accumulating evidence for convergent validity. This article introduces a Current Issues section that gives new attention to validity concerns associated with student ratings. The section's 4 articles deal, respectively, with (a) conceptual structure (are student ratings unidimensional or multidimensional?), (b) convergent validity (how well do ratings correlate with other indicators of effective teaching?), (c) discriminant validity (are ratings influenced by factors other than teaching effectiveness?), and (d) consequential validity (are ratings used effectively in personnel development and evaluation?). Although all 4 articles favor the use of ratings, they disagree on controversial points associated with interpretation and use of ratings data. PMID- 9357334 TI - Can preventive gerontology be on the way? PMID- 9357335 TI - Goliath and some Davids in the tobacco wars. PMID- 9357336 TI - Issues in equalizing Medicare expenditures--the devil is in the details. PMID- 9357337 TI - The knowledge base for public health strategies. PMID- 9357339 TI - From public health science to prevention policy: placing science in its social and political contexts. PMID- 9357338 TI - Ethical challenges posed by clinical progress in AIDS. PMID- 9357340 TI - The invisibility of public health: population-level measures in a politics of market individualism. AB - This paper models the prevailing political attack on government as a heuristic, a judgmental strategy that simplifies complex phenomena by applying simple tests to a limited set of relevant data. The heuristic of market individualism offers three tools for analyzing the problems of governing: the supremacy of the free market as a regulatory device, a belief in individual freedom of choice and personal responsibility, and the elevation of individual satisfaction as the chief goal of society. Because public health is inherently concerned with the health of the population rather than individual health, because the market itself is a major source of ill health, and because individual choice is socially mediated, use of the heuristic precludes the recognition of the value of public health work. Although some degree of accommodation to current political realities is a practical necessity, public health advocates must not neglect the task of fashioning radically different alternative visions over the long term. PMID- 9357341 TI - State variations in Medicare expenditures. AB - OBJECTIVES: This study examined variations in Medicare expenditures across states. METHODS: 1992 data on average Medicare expenditures per enrollee, users of services per 1000 enrollees, service use per user, and payment per unit of service were compared across states for various services. Weighted least squares regression analysis was employed to examine total Medicare expenditures per enrollee by state. RESULTS: Variation in Medicare expenditures across states is driven more by average number of service users per 1000 enrollees and average service units per user than by average payment per service unit. Medicare expenditures per enrollee by state are primarily a function of Medicare HMO penetration rate (P = .000), urban area (P = .001), hospital bed supply (P = .005), elderly mortality rate (P = .012), Medicare physician assignment rate (P = .026), percentage of primary care practitioners (P = .042), and interactions between urban elderly and percentage of primary care physicians (P = .005) and Black elderly and nursing home bed supply (P = .012). CONCLUSIONS: Before sweeping Medicare cuts are undertaken or excessive reliance on managed care occurs, attention should be focused on the current disproportionate distribution of expenditures across states. PMID- 9357343 TI - Prescription drug spending: the impact of age and chronic disease status. AB - OBJECTIVES: The purpose of this study was to examine how pharmaceutical expenditures vary by age and the presence of chronic health problems. METHODS: Data from the 1987 National Medical Expenditure Survey were used to obtain nationally representative estimates of outpatient prescription drug expenditures for the noninstitutionalized population and the fraction of total health expenditures used to purchase medications for age-chronic disease population subgroups. RESULTS: Although the elderly make up 12% of the population, they account for 34% of total pharmaceutical expenditures. Pharmaceutical expenditures are 9% of total expenditures for children, 13% for nonelderly adults, and 23% for the elderly. Among nonelderly adults, approximately one third have at least one chronic condition and account for over two thirds of drug expenditures. Among the elderly, 36% have three or more chronic conditions and account for 57% of drug expenditures for this group; 41% of total drug expenditures are for cardiovascular or renal drugs. CONCLUSIONS: Significant pharmaceutical spending is for treatment of chronic conditions, which subjects insurance coverage to adverse selection and could affect the design of prescription drug benefit packages. Current enrollees in Medicare risk management plans who have drug benefits may face significantly higher out-of-pocket expenses for pharmaceuticals if capitation rates are cut as a means of controlling Medicare program expenditures. PMID- 9357342 TI - Age, time, and cohort effects on functional status and self-rated health in elderly men. AB - OBJECTIVES: This study investigated age-related changes in functional status and self-rated health in elderly men, taking into account changes over time and differences between birth cohorts. METHODS: The Zutphen Elderly Study is a longitudinal study of men born in the Netherlands between 1900 and 1920. Functional status and self-rated health were measured in 513 men in 1990, in 381 men in 1993, and in 340 men in 1995. Age, time, and cohort effects were analyzed in a mixed longitudinal model. RESULTS: Longitudinal analyses showed that during 5 years of follow-up, the proportion of men without disabilities decreased from 53% to 39%, whereas the percentage who rated themselves as healthy decreased from 50% to 35%. Cross-sectional analyses confirmed changes in functional status, suggesting an age effect. Time-series analyses confirmed changes in self-rated health, suggesting a time effect. No birth-cohort effects were found. CONCLUSIONS: Functional status deteriorates with age, whereas self-rated health is not related to age in men aged 70 years and older. The observed 5-year decline in self-rated health seemed to be due to a secular trend. PMID- 9357344 TI - Excess mortality attributable to hip fracture in white women aged 70 years and older. AB - OBJECTIVE: The purpose of this study was to estimate the excess mortality attributable to hip fracture. METHODS: The 6-year survival rate of community dwelling White female hip fracture patients aged 70 years and older entering one of seven hospitals from 1984 to 1986 (n = 578) was compared with that of White female respondents aged 70 years and older interviewed in 1984 for the Longitudinal Study on Aging (n = 3773). RESULTS: After age, education, comorbidity, and functional impairment were controlled, the mortality differential between the two groups accumulated to an excess among hip fracture patients of 9 deaths per 100 women 5 years postfracture. Among those with three or more functional impairments or one or more comorbidities, the excess was 7 deaths per 100: the effect of the fracture had disappeared in these groups by 4 years. In contrast, those with two or fewer impairments and those with no comorbidities had a continuing trend of increased mortality, with an excess of 14 deaths per 100 by 5 years. CONCLUSIONS: There is an immediate increase in mortality following a hip fracture in medically ill and functionally impaired patients, whereas among those with no comorbidities and few impairments, there is a gradual increase in mortality that continues for 5 years postfracture. PMID- 9357345 TI - The effects of planned duration of residential drug abuse treatment on recovery and HIV risk behavior. AB - OBJECTIVE: This study assessed the effects of planned duration of residential drug abuse treatment on recovery from drug use and on human immunodeficiency virus (HIV) risk behaviors. METHODS: Two concurrent randomized controlled trials of programs differing in planned duration were conducted: 6-month vs 12-month versions of a traditional therapeutic community program, and 3-month vs 6-month versions of a modified therapeutic community incorporating a relapse prevention and health education program. Outcomes, measured at least 16.5 months after admission, included time from admission to first drug use; severity of drug, alcohol, legal, and employment problems; and risky drug injection and sexual behaviors. RESULTS: Among 539 clients (86% of those enrolled), there were no significant effects of planned duration of treatment upon Addiction Severity Index scores or HIV risk behavior. In the relapse prevention program, clients randomized to the 6-month program had a longer time to first drug use than those in the 3-month program (hazard ratio = 0.74; 95% confidence interval = 0.58, 0.93). Employment problems at follow-up were significantly less severe among clients treated in the therapeutic community than among those in the relapse prevention program. CONCLUSIONS: No overall benefit of extending treatment beyond 6 months was found. PMID- 9357346 TI - Are the best coronary artery bypass surgeons identified by physician surveys? AB - OBJECTIVES: This study assessed the validity of surveys for identifying the best coronary artery bypass surgeons. METHODS: Data on physicians who performed coronary artery bypass surgery were available from New York, Pennsylvania, and Wisconsin. Data on physicians' reputation were obtained from one national and five city surveys. The measure of surgical performance was the mortality ratio (MR), that is, the ratio of the observed to the predicted patient mortality rate. RESULTS: Mortality ratios were very similar for the 10,722 patients treated by the 31 surgeons defined as "best" doctors in the surveys (MR = 98) and for the 74,854 patients treated by 243 other surgeons who had more than a minimal number of cases (MR = .96). The mortality ratio was 1.34 for the patients treated by surgeons with the lowest volumes and .87 for the surgeons who performed more than 400 coronary artery bypass surgeries in 3 years. CONCLUSIONS: These results suggest that the quality of a coronary artery bypass surgeon may be more closely associated with patient volume than with the surgeon's reputation among peers. PMID- 9357347 TI - Daytime sleepiness: an epidemiological study of young adults. AB - OBJECTIVES: Although excessive daytime sleepiness is associated with increased risks for accidents, decreased productivity, and interpersonal difficulties, information on its epidemiology is scarce. This paper examines correlates of and suspected risk factors for daytime sleepiness from a longitudinal epidemiological study of young adults. METHODS: The sample consisted of 1007 randomly selected young adults from a large health maintenance organization in southeast Michigan. Data were gathered in personal interviews conducted with 97% of the sample 5.5 years after baseline. Information on sleep characteristics in the last 2 weeks, including daytime sleepiness, nocturnal sleep onset, snoring, and hours of sleep, was collected on a self-administered instrument. Psychiatric disorders were measured by the National Institute of Mental Health's Diagnostic Interview Schedule. RESULTS: The average length of nocturnal sleep on weekdays was 6.7 hours. Daytime sleepiness was inversely related to hours of sleep and positively related to the ease of falling asleep at night; it varied significantly by employment and marital status. Snoring was associated with increased daytime sleepiness, as was recent major depression. CONCLUSIONS: Factors that might increase daytime sleepiness among young adults include social factors (being single and being employed full time) and pathological conditions (frequent snoring and major depression). PMID- 9357348 TI - The changing distribution of HIV infection: HIV surveillance in Lazio, Italy, 1985 through 1994. Lazio HIV Surveillance Collaborative Group. AB - OBJECTIVES: This study sought to describe the human immunodeficiency virus (HIV) surveillance system in Lazio, Italy, and to analyze exposure patterns and time trends of HIV serodiagnoses from January 1985 to December 1994. METHODS: A linkage procedure made it possible to identify newly diagnosed HIV cases. Anonymous information was collected on demographic and exposure factors for each individual. RESULTS: Of 35,425 reports, 13,660 were newly diagnosed HIV cases, 70.9% of them in men. The proportion of women increased at the beginning of the study period (the male:female ratio declined from 3.5 in 1985 to 2.6 in 1986) and then remained stable. The proportion of subjects reporting heterosexual exposure, in men and women, respectively, increased from 1.5% and 2.0% in 1985 to 21.2% and 60.8% in 1994. Starting in 1992, heterosexual contact has become the main transmission route for women. CONCLUSIONS: A changing pattern in the HIV epidemic is emerging, with a shift in the incidence of HIV diagnosis from "core" high-risk groups (drug injectors) to the large low-risk population (the general population) exposed through heterosexual transmission. This is probably occurring in other areas (e.g., large urban centers in the United States) with a similar epidemiological situation. PMID- 9357349 TI - Improving publicly funded substance abuse treatment: the value of case management. AB - OBJECTIVES: This study evaluated the impact of case management on client retention in treatment and short-term relapse for clients in the publicly funded substance abuse treatment system. METHODS: A retrospective cohort design was used to study clients discharged from the following four modalities in 1993 and 1994: short-term residential (3112 clients), long-term residential (2888 clients), outpatient (7431 clients), and residential detox (7776 clients). Logistic regression models were used to analyze the impact of case management after controlling for baseline characteristics. RESULTS: The odds that case-managed clients reached a length of stay previously identified as associated with more successful treatment were 1.6 (outpatient programs) to 3.6 (short-term residential programs) times higher than the odds for non-case-managed clients. With the exception of outpatient clients, the odds of case-managed clients' being admitted to detox within 90 days after discharge (suggesting relapse) were about two thirds those of non-case-managed clients. The odds of case-managed detox clients' transitioning to post-detox treatment (a good outcome) were 1.7 times higher than the odds for non-case-managed clients. CONCLUSIONS: Case management is a low-cost enhancement that improves short-term outcomes of substance abuse treatment programs. PMID- 9357350 TI - State smoking prevalence estimates: a comparison of the Behavioral Risk Factor Surveillance System and current population surveys. AB - OBJECTIVES: This study examined whether there are systematic differences between the Behavioral Risk Factor Surveillance System (BRFSS) and the Current Population Survey (CPS) for state cigarette smoking prevalence estimates. METHODS: BRFSS telephone survey estimates were compared with estimates from the US Census CPS tobacco-use supplements (the CPS sample frame includes persons in households without telephones). Weighted overall and sex- and race-specific BRFSS and CPS state estimates of adults smoking were analyzed for 1985, 1989, and 1992/1993. RESULTS: Overall estimates of smoking prevalence from the BRFSS were slightly lower than estimates from CPS (median difference: -2.0 percentage points in 1985, -0.7 in 1989, and -1.9 in 1992/1993; P < .05 for all comparisons), but there was variation among states. Differences between BRFSS and CPS estimates were larger among men than among women and larger among Blacks than among Hispanics or Whites; for most states, these differences were not significant. CONCLUSIONS: The BRFSS generally provides state estimates of smoking prevalence similar to those obtained from CPS, and these are appropriate for ongoing state surveillance of smoking prevalence. PMID- 9357352 TI - Injury and death associated with hospital bed side-rails: reports to the US Food and Drug Administration from 1985 to 1995. AB - OBJECTIVES: Hospital bed side-rails, while intended for patient protection, can contribute to injury and death. Reports to the Food and Drug Administration (FDA) of hospital bed side-rail entrapment have increased. In this paper entrapment cases are reviewed and the population potentially at risk identified. METHODS: FDA's database was searched for events involving hospital beds from January 1985 to August 1995 and entrapment cases were identified. RESULTS: Of 111 entrapments, 65% were associated with death and 23% with injury. CONCLUSIONS: Advanced age, female sex, low body weight, and cognitive impairment may be associated with increased risk. Preventive measures are detailed. PMID- 9357351 TI - Telephone support as an adjunct to transdermal nicotine in smoking cessation. AB - OBJECTIVES: Transdermal nicotine patches have shown considerable promise in improving smoking cessation outcomes. The present study assessed telephone support as an adjunct to a managed care-based, single-session group orientation smoking cessation program with nicotine patch therapy. METHODS: The unit of randomization was the orientation session (n = 35). Subjects (n = 509) were randomly assigned to a group session without telephone support, the session plus access to a toll-free help line, or the session with telephone help line plus active telephone outreach. RESULTS: Contrary to hypothesis, there were no differences between treatment conditions. Overall abstinence rates were 22% at 6 months and 21% at 1 year. Fewer than 1% of eligible subjects called the toll-free help line. An average of 3.8 of a possible 4 calls were completed in the telephone outreach condition. CONCLUSIONS: Abstinence results obtained in this program were comparable to those obtained with more extensive counseling. However, there was no evidence of benefit from telephone support beyond the initial physician-led group orientation session. PMID- 9357353 TI - Consequences of foot binding among older women in Beijing, China. AB - OBJECTIVES: This study examined the prevalence and consequences of foot binding in older Chinese women. METHODS: Women older than 70 years in Beijing, China, were assessed for bound feet, falls, functional status, and bone density. RESULTS: Thirty-eight percent of women aged 80 years and older and 18% of women aged 70 through 79 years had bound-foot deformities. Women with bound feet were more likely to fall, less able to squat, and less able to stand up from a chair without assistance than women with normal feet. They also had 14.3% less functional reach (a test of balance) and 5.1% lower hip bone density. CONCLUSIONS: Foot binding has caused substantial disability that is still evident in many elderly Chinese women. PMID- 9357354 TI - The contribution of six chronic conditions to the total burden of mobility disability in the Dutch population. AB - OBJECTIVES: This study assessed the proportions of the burden of mobility disability in the Dutch population that are attributable to musculoskeletal diseases, lung diseases, neurological disorders, heart diseases, diabetes, and cancer. METHODS: National survey data were analyzed with an elimination technique that combines the results of logistic regression analysis and the disease prevalence. RESULTS: Of the total prevalence of disability (20.5%), 33.7% can be attributed to these six chronic conditions. Musculoskeletal disorders account for the major part, whereas the contribution of cancer is very small. CONCLUSIONS: The potential benefits of effective curative or preventive treatments for chronic conditions, in terms of reduction of the disability burden in the population, are limited. PMID- 9357356 TI - The effect of ordinances requiring smoke-free restaurants and bars on revenues: a follow-up. AB - OBJECTIVES: The purpose of this study was to extend an earlier evaluation of the economic effects of ordinances requiring smoke-free restaurants and bars. METHODS: Sales tax data for 15 cities with smoke-free restaurant ordinances, 5 cities and 2 counties with smoke-free bar ordinances, and matched comparison locations were analyzed by multiple regression, including time and a dummy variable for the ordinance. RESULTS: Ordinances had no significant effect on the fraction of total retail sales that went to eating and drinking places or on the ratio between sales in communities with ordinances and sales in comparison communities. Ordinances requiring smoke-free bars had no significant effect on the fraction of revenues going to eating and drinking places that serve all types of liquor. CONCLUSIONS: Smoke-free ordinances do not adversely affect either restaurant or bar sales. PMID- 9357355 TI - A two-step intervention of increase mammography among women aged 65 and older. AB - OBJECTIVES: This study evaluated a two-step intervention for mammography screening among older women. METHODS: Four hundred and sixty women, identified from physician practices, were randomized to a control or a two-step intervention (physician letter and peer counseling call) group. Women in the intervention group who obtained a mammogram received a grocery coupon. RESULTS: Over the 12 months of the study, more women in the intervention group than in the control group obtained mammograms (38% vs 16%). The most dramatic difference was in the higher odds that women in the intervention group would obtain a mammogram within 2 months (odds ratio = 10.5). CONCLUSIONS: The intervention significantly increased screening mammography. Future efforts must be multifaceted and incorporate the unique concerns of older women. PMID- 9357357 TI - Thyroxine values from newborn screening of 919 infants born before 29 weeks' gestation. AB - OBJECTIVES: Severe transient hypothyroxinemia in premature infants is associated with cerebral palsy and mental retardation: this study assessed its prevalence in very premature infants. METHODS: Congenital hypothyroidism screening programs in three states provided thyroxine values for 919 newborn infants younger than 29 weeks who were enrolled in a multicenter study. RESULTS: Thyroxine values were lower than 4.0 micrograms/dL in 21% of survivors and increased each week by 0.6 microgram/dL (95% confidence interval [CI] = 0.4, 0.7). At tests done 1 to 2 days after birth, levels were 2.5 micrograms/dL higher (95% CI = 1.8, 3.3) than at tests done at 8 to 14 days. In New York, levels were 1.0 microgram/dL higher (95% CI = 0.3, 1.6) than elsewhere. The levels of infants who died were 1.3 micrograms/dL lower (95% CI = 0.6, 2.0) than those of survivors. CONCLUSIONS: Severe transient hypothyroxinemia is common in very premature infants and deserves further study. PMID- 9357358 TI - Residential lead-based-paint hazard remediation and soil lead abatement: their impact among children with mildly elevated blood lead levels. AB - OBJECTIVES: This prospective study describes the impact of residential lead-based paint hazard remediations on children with mildly elevated blood lead levels. METHODS: Changes in blood lead levels were observed following paint hazard remediation alone and in combination with soil abatement. RESULTS: After adjustment for the confounding variables paint hazard remediation alone was associated with a blood lead increase of 6.5 micrograms/dL (P = 0.5), and paint hazard remediation combined with soil abatement was associated with an increase of 0.9 microgram/dL (P = 36). CONCLUSIONS: Lead-based-paint hazard remediation as performed in this study, is not an effective secondary prevention strategy among children with mildly elevated blood lead levels. PMID- 9357359 TI - Dog bite incidence in the city of Pittsburgh: a capture-recapture approach. AB - OBJECTIVES: The purpose of this study was to estimate the number of dog bite injuries occurring in the city of Pittsburgh in 1993. METHODS: The capture recapture method was used, along with long-linear modeling. Three sources were used to identify victims hospital reports, animal control reports, and police/victim reports. RESULTS: In 1993, 790 dog bites were reported. The capture recapture method estimated that there were 1388 unreported dog bites, with an estimated incidence rate of 58.9 per 10,000. CONCLUSIONS: Dog bite is a common our preventable injury. To improve surveillance, the focus should be on educating the general public about the serious consequences of dog bite injuries. PMID- 9357361 TI - Unintended pregnancy and breast-feeding behavior. AB - OBJECTIVES: This study assessed the effect of unintended pregnancy on breast feeding behavior. METHODS: All women delivering a live birth between January 1, 1995, and July 31, 1996 (n = 33,735), in the 15-county central New York region were asked whether they had intended to become pregnant and their breast-feeding plans. RESULTS: Women with mistimed pregnancies, and pregnancies that were not wanted were significantly less likely to breast-feed than were women whose pregnancies were planned. After adjustment for confounding variables and contraindications for breast-feeding, the odds ratios of not breast-feeding remained significant. CONCLUSIONS: Promoting breast-feeding among women with unintended pregnancies is important to improve health status. PMID- 9357360 TI - 'Call fast, Call 911': a direct mail campaign to reduce patient delay in acute myocardial infarction. AB - OBJECTIVES: A 10-month direct mail campaign was implemented to increase use of emergency medical services via 911 calls and to reduce prehospital delay for individuals experiencing acute myocardial infarction symptoms. METHODS: This prospective, randomized, controlled trial involved three intervention groups (receiving brochures with informational, emotional, or social messages) and a control group. RESULTS: Intervention effects were not observed except for individuals who had a history of acute myocardial infarction and who were discharged with a diagnosis of acute myocardial infarction; their 911 use was meaningfully higher in each intervention group than in the control group. CONCLUSIONS: The mailings affected only the individuals at greatest risk. PMID- 9357363 TI - Hepatitis A among schoolchildren in a US-Mexico border community. AB - OBJECTIVES: A cross-sectional study investigated the association of hepatitis A seropositivity with environmental and personal risk factors among children in a United States-Mexico border community. METHODS: Hepatitis A serological markers and a questionnaire identifying risk factors were evaluated for 523 primary school children. RESULTS: Of the children studied, 16.9% tested positive for total antihepatitis A virus. Risk factors included being in the first grade, low maternal educational attainment, living in Mexico for more than 6 months, household crowding, and inadequate excreta disposal systems. CONCLUSIONS: To decrease enteric disease, improvements in excreta disposal infrastructures and educational programs are needed. Hepatitis A vaccine should be administered before school age. PMID- 9357364 TI - Diabetes in Hawaii: estimating prevalence from insurance claims data. AB - OBJECTIVES: The purpose of this study was to determine the prevalence of diabetes mellitus in Hawaii from insurance claims data. METHODS: Information from two major health plans covering approximately 66% of the state's population was used to estimate prevalence rates by sex, age group, and geographic area. Weighted multiple linear regression was applied to identify predictors of diabetes prevalence. RESULTS: The statewide diabetes prevalence was estimated at 43.8 per 1000 persons. The ethnic composition of the population and rural residence partially explained the geographic variation in diabetes prevalence. CONCLUSIONS: Insurance claims data may be a useful tool for population-based diabetes surveillance. PMID- 9357362 TI - Plasma polychlorinated biphenyl levels in Dutch preschool children either breast fed or formula-fed during infancy. AB - OBJECTIVES: This study examined the influence of lactational and in utero exposure to polychlorinated biphenyls (PCBs) on plasma PCB levels in children. METHODS: Plasma PCB levels were measured in 173 children at 3.5 years, of whom 91 were breast-fed and 82 were formula-fed in infancy. RESULTS: Median plasma PCB levels were 3.6 times higher in breast-fed children (0.75 microgram/L) than in their formula-fed peers (0.21 microgram/L). Breast-feeding period and breast-milk PCB levels were important predictors for PCB levels in the breast-fed group. For children in the formula-fed group, PCB levels were significantly related to their material plasma PCB levels. CONCLUSIONS: PCB levels in Dutch preschool children are related to transfer of maternal PCBs; therefore, strategies should be aimed at reducing maternal PCB body burden. PMID- 9357365 TI - An investigation of increased tuberculosis case reports in Santa Clara County, California, 1993-1994. PMID- 9357367 TI - Hepatitis C virus infection among Alaskan drug users. PMID- 9357366 TI - On 'accidents'. PMID- 9357368 TI - Lead and other metals in play kit and craft items composed of vinyl and leather. PMID- 9357369 TI - Do we ask too much from community-level interventions or from intervention researchers? PMID- 9357371 TI - Human immunodeficiency virus infection in a patient with primary immunodeficiency. PMID- 9357370 TI - Endotoxin: a role in latex allergy? PMID- 9357372 TI - Sick building syndrome--a wolf in sheep's clothing. AB - OBJECTIVE: Reading this article will acquaint the reader with possible outcomes associated with the diagnosis of "sick building syndrome." The definition, epidemiology, and precipitating events of this symptom complex are distinguished from other defined building-related illnesses. DATA SOURCE: The author's experience with many patients presenting with this diagnostic label and selected studies on indoor pollution and "sick building syndrome" are carefully reviewed. STUDY SELECTION: Pertinent scientific investigations on "sick building syndrome" and previously published reviews on this and related subjects that met the educational objectives were critically reviewed. RESULTS: "Sick building syndrome" is a pseudodiagnosis composed of nonspecific, transient symptoms without proven biologic markers. Its application in the clinical setting invites frequent subsequent linkage to other similar vague diagnoses associated with chronic debility and lack of therapeutic intervention. CONCLUSION: The reader is encouraged to avoid the use of this term in favor of a simpler, descriptive diagnosis (e.g., transient office-related annoyance and/or irritation) or if this seems inadequate, adoption of the diagnostic label of "idiopathic building intolerance." PMID- 9357373 TI - Progressive immunodeficiency in a patient with IgA deficiency. PMID- 9357374 TI - Endotoxin as a factor in adverse reactions to latex gloves. AB - BACKGROUND: Endotoxin is an inflammatory made by gram negative bacteria that can irritate the skin, induce respiratory problems, fever, and shock. It is an adjuvant for both delayed hypersensitivity and IgE production and has been shown to magnify antigen specific mediator release. Since many of the clinical problems associated with natural latex products involve similar clinical sequelae, we investigated the possibility that latex gloves might be contaminated with endotoxin. OBJECTIVE: To measure the endotoxin content of a variety of natural latex gloves, investigate the its distribution and origin, associated with latex proteins, and determine the particle sizes associated with its release. METHODS: Endotoxin, protein, and allergen were measured using a quantitative kinetic Limulus assay, modified Lowry, and RAST inhibition, respectively. Particle size and density were determined using an Anderson multistage air sampler and CsCl2 gradient. RESULTS: Endotoxin was found to be a highly significant contaminant of some latex gloves. Levels ranged from 0.09 ng to 2.8 micrograms/g of glove. Protein levels ranged from < 25 to 1150 micrograms/g of glove while allergen levels ranged from < 1 to 837 micrograms/g of glove. Endotoxin and protein eluted rapidly from the interior of the gloves tested. Greater than 70% of the endotoxin was found to be associated with particles in the < 7 microns aerodynamic diameter range. The highest levels of endotoxin were found in nonsterile examination gloves with a tendency towards powdered gloves containing more endotoxin and protein. A slurry containing cross-linked dextran through which gloves were dipped revealed very high endotoxin contamination (64 micrograms/mL) while unused cross-linked dextran has very little associated endotoxin. CONCLUSIONS: These data demonstrate that some natural rubber latex gloves, particularly nonsterile examination gloves, are contaminated with high amounts of endotoxin and proteins. These were found mostly on the inside of gloves and were released as very small respirable particles that were not physically associated with the powder. These findings support the hypothesis that endotoxin may be responsible for some of the tissue irritation associated with latex glove use. In addition, this material may be responsible for the enhancement of delayed and immediate hypersensitivity reactions to chemicals and proteins found in these products and offers a possible explanation for the disproportionate severity of these reactions. PMID- 9357375 TI - Maintaining theophylline compliance/adherence in severely asthmatic children: the role of psychologic functioning of the child and family. AB - BACKGROUND: Noncompliance with asthma therapy by asthmatic children and their families is a common cause of treatment failure. Psychologic factors have been reported to influence adherence patterns. OBJECTIVE: To evaluate the relationship of psychologic function of severely asthmatic children and families with theophylline compliance during a 1-year follow-up period after inpatient asthma rehabilitation. METHOD: Thirty-seven severely asthmatic children with a median age of 9 years (range, 2 to 17 years) were treated in an inpatient rehabilitation program (median, 15 days). Established clinical rating scales of psychologic adjustment were used for children (Child Global Assessment Scale) and families (Family Global Assessment Scale) at the time of admission. Patients were defined as noncompliant if 30% or more of theophylline levels obtained during the 1-year follow-up period were less than 5 mg/L (mumol/L). RESULTS: Compliant children had higher scores on the Child Global Assessment Scale than noncompliant children (64 versus 48, P = .09). Families of compliant children had significantly higher scores on the Family Global Assessment Scale than did families with noncompliant children (63 versus 54, P < .05). There was no statistical relationship between compliance status and parent-reported behavior problems, demographic factors (age, race, sex, and number of parents at home), health insurance, specialty referral at admission, measures of medical morbidity (days hospitalized, number of emergency care visits, and number of corticosteroid bursts), or length of rehabilitation stay. Both compliant and noncompliant children had comparable reductions in morbidity (hospital and emergency care, number of corticosteroid bursts required). CONCLUSION: Psychologic functioning of the child and family may be related to theophylline compliance. PMID- 9357376 TI - Paracetamol (acetaminophen) hypersensitivity. AB - BACKGROUND: Paracetamol (acetaminophen) intolerance can occur in less than 5% of aspirin-sensitive subjects as a result of inhibition of prostaglandin synthesis but hypersensitivity to paracetamol without aspirin sensitivity is rare. METHODS: We report a case of an acute life-threatening hypersensitivity reaction to paracetamol. RESULTS: An oral challenge with 125 mg of paracetamol was carried out and followed by generalized anaphylactic reaction. An oral provocation test with 500 mg aspirin was well tolerated in the patient. CONCLUSION: An acute hypersensitivity reaction to paracetamol is described in this study. The results of the study suggest a real allergic mechanism rather than inhibition of prostaglandin synthesis as responsible. PMID- 9357378 TI - Azelastine nasal spray as adjunctive therapy to azelastine tablets in the management of seasonal allergic rhinitis. AB - BACKGROUND: Azelastine rhinitis medications (nasal spray and tablets) have been shown to relieve the symptoms of allergic rhinitis. Nevertheless, many rhinitic subjects suffer from acute exacerbations of symptoms that sometimes require additional treatment. OBJECTIVE: To assess the efficacy and safety of azelastine nasal spray as adjunctive therapy to azelastine tablets in the management of symptomatic seasonal allergic rhinitis in subjects who remain symptomatic despite the oral medication. METHODS: A 2-day, randomized, multicenter, double-blind, placebo-controlled, parallel-group study. Two hundred thirty-three subjects with symptomatic allergic rhinitis received azelastine tablets (0.5 mg bid) for a minimum of seven days prior to receiving either azelastine nasal spray (2 sprays per nostril bid) or placebo nasal spray as adjunctive therapy. Efficacy was determined by improvement in rhinitis symptoms that were grouped according to total and major symptom complex severity scores. RESULTS: Mean percent improvements in the total symptom complex severity scores for azelastine were statistically significant (P < or = .05) or showed a trend toward statistical significance (.05 < or = P < .10) versus placebo from the second through the first ten hours after the initial dose and for each of the last five hours of the second day, demonstrating a rapid onset of action and sustained efficacy over the 2-day study period. Azelastine was well tolerated, and no subject discontinued therapy with azelastine due to an adverse experience. CONCLUSION: Azelastine nasal spray can be effectively administered as adjunctive therapy, in an outdoor environment in which subjects are exposed to pollen and other aeroallergens. PMID- 9357377 TI - Pilot study of bronchodilator response to inhaled albuterol delivered by metered dose inhaler and a novel dry powder inhaler. AB - BACKGROUND: The metered-dose inhaler is currently one of the most prescribed methods of delivering drugs to the lungs. In the United States, most currently marketed metered dose inhalers use chlorofluorocarbons as the system propellant and require patient breath coordination. These factors lead to the need for a delivery system that is independent of propellants and patient coordination. OBJECTIVE: To compare the magnitude and time course of bronchodilation between albuterol delivered by Ventolin metered dose inhaler and albuterol sulfate powder (Rotacaps) delivered by a novel dry powder inhaler that generates a respirable drug aerosol over a range of inspiratory flow rates. METHODS: A single-center, single-dose, randomized, placebo-controlled, partial-blind, 3-way crossover study was conducted in an outpatient asthma Clinical Research Center. Twelve mild to moderate asthmatic patients 12 to 36 years of age participated in this study that involved three treatments, each separated by three to eight days, consisting of 2 puffs (90 micrograms/puff) albuterol by Ventolin metered-dose inhaler, two inhalations (100 micrograms/puff) albuterol sulfate powder (Rotacaps) by dry powder inhaler, and two inhalations (12.5 mg/inhalation) lactose powder by dry powder inhaler. Spirometry, blood pressure, and heart rate were measured at 30 minutes, 15 minutes, and immediately before treatment and then at 15, 30, 45, 60, 90, 120, 180, 240, and 300 minutes after each treatment. Serum potassium and glucose, and electrocardiograms were measured at 30 minutes before, and 30, 60, 90, and 180 minutes after each treatment. Endpoints were compared with analysis of variance. RESULTS: Five patients (one metered-dose inhaler and four dry powder inhaler) did not respond with > 15% FEV1 increase over baseline within 30 minutes. Metered-dose inhaler and dry powder inhaler mean FEV1 results, respectively, for 11 and 8 responders were 15 minutes in onset, 202.9 and 185.4 minutes in duration, 24.8% and 25.1% maximum change, and 18.6 and 18.2 area-under FEV1-bronchodilation-curve. Statistical analysis of all patients and responders only revealed both active treatments to be different from placebo (P = .0018), but not different from each other (P = .1291). No safety endpoints were significantly different among all three treatments (P > .10 for all safety endpoints). CONCLUSIONS: In this study, the dry powder inhaler safely and effectively delivered a commercially available albuterol sulfate powder (Rotacaps) into human lungs with bronchodilation comparable to Ventolin metered dose inhaler. PMID- 9357379 TI - Esophageal candidiasis as a complication of inhaled corticosteroids. AB - BACKGROUND: Oropharyngeal candidiasis is a well-described side effect of inhaled corticosteroids. Nevertheless, few cases of esophageal candidiasis have been reported. OBJECTIVE: To present a patient with esophageal candidiasis associated with inhaled corticosteroids. METHODS: Case report. RESULTS: Our patient is a 70 year-old white woman with a 20-year history of intrinsic asthma, well controlled on triamcinolone acetonide 400 micrograms, ipratropium bromide 36 micrograms, and pirbuterol acetate 400 micrograms, each inhaled four times daily. She reported no oral steroid use for > 4 years and that she always rinsed her mouth following triamcinolone acetonide inhalation. The patient had gastritis with peptic ulcer disease in the past and developed worsening dyspeptic pain and heartburn. Following discontinuation of cimetidine and initiation of ranitidine without improvement, esophagogastroduodenoscopy was performed. Several small white patches in the mid and distal esophagus could not be removed with pressure. A biopsy confirmed the diagnosis of candidal esophagitis. Following a 4-week course of fluconazole, the patient was clinically improved and follow-up esophagogastroduodenoscopy was normal. There was no evidence of underlying cellular immunosuppression, malignancy, or diabetes mellitus and no history of recent antibiotic usage. Delayed skin tests revealed 5 x 5 mm induration to dermatophytin. Delayed hypersensitivity to Candida and mumps tests was absent. There was strong in vitro lymphocyte transformation and a positive immediate skin test response to Candida. ELISA for human immunodeficiency virus was negative. T and B cell counts were normal with CD4 = 630/mm3, CD8 = 520/mm3, and absolute B cell = 120/mm3. It is possible that this patient's immediate hypersensitivity response to Candida suppressed her delayed response. Candidal esophagitis is a rare, yet important, complication of inhaled corticosteroid use. CONCLUSION: Immunocompetent patients on inhaled corticosteroids with medically unresponsive symptoms of esophagitis should be investigated for esophageal candidiasis. PMID- 9357380 TI - Mugwort and sage (Artemisia) pollen cross-reactivity: ELISA inhibition and immunoblot evaluation. AB - BACKGROUND: Plants of the genus Artemisia are a source of fall allergic symptoms, particularly in the western United States. Studies have characterized the allergens in one of the major species (A. vulgaris) but currently there are no cross-reactivity data on the major United States species. OBJECTIVE: The purpose of this study was to investigate the in vitro cross-reactivity among nine Artemisia species: A. frigida, A. annua, A. biennis, A. filifolia, A. tridentata, A. californica, A. gnaphalodes, A. ludoviciana, and A. vulgaris. METHODS: The cross-reactivity was demonstrated with the use of enzyme-linked immunosorbent assay (ELISA) inhibitions and immunoblotting techniques utilizing a serum pool from patients allergic to Artemisia species. RESULTS: The enzyme-linked immunosorbent assay inhibitions revealed strong cross-reactivity among all nine species with A. biennis and A. tridentata being two of the strongest inhibitors. The polyacrylamide gel electrophoresis showed a great deal of similarity in the bands among the nine species. The nitrocellulose blots showed similar IgE binding patterns among the Artemisia species with strong inhibition among all nine extracts. CONCLUSIONS: These data all demonstrate very strong in vitro cross reactivity among the nine Artemisia species studied. Such data have significant clinical relevance, suggesting that a single Artemisia species may be sufficient for allergy skin testing and formulation of immunotherapy extracts. PMID- 9357381 TI - Tourette's syndrome in patients referred for allergy evaluation. AB - BACKGROUND: Tourette's syndrome is a relatively common disorder that may present as a spectrum that can include both motor and behavioral features. METHODS: We report four patients referred to allergists to rule out atopy as a reason fro their presumed respiratory symptoms who were subsequently found to have Tourette's syndrome. A review of the literature was also performed using Medline. RESULTS: The symptoms of Tourette's syndrome may overlap those associated with allergy involving the respiratory tract. There is little published information on the association of Tourette's syndrome with atopy. CONCLUSIONS: Allergists should be aware of Tourette's syndrome and the variable clinical spectrum of this disorder as Tourette's syndrome patients may be referred prior to the diagnosis of Tourette's syndrome. PMID- 9357382 TI - Unsuspected sources of soybean exposure. AB - BACKGROUND: Hidden allergens in processed foods can provoke severe allergic reactions in sensitive individuals. The presence of soy proteins in typical Spanish sausage products (chorizo, salchichon, mortadella, and boiled ham), doughnut and soup stock cubes has not been reported previously. METHODS: The present article reports two examples of severe allergic reactions due to the ingestion of foods that unexpectedly contained soybean proteins. Allergollogic study included skin prick tests with the implicated foods and their components, serum specific IgE and bronchial and oral provocation tests. RESULTS: Skin test, serum-specific IgE, and bronchial and oral challenge tests (the latter, in one patient) were positive against soy and the above mentioned processed foods in which the presence of soybean flour was confirmed. CONCLUSIONS: This report demonstrates the importance of hidden allergens in allergic reactions to foods and the need to scrutinize closely every food component. PMID- 9357383 TI - Demographic predictors of asthma treatment site: outpatient, inpatient, or emergency department. AB - OBJECTIVE: To identify the demographic predictors of asthma treatment site: outpatient clinic, emergency department, or hospital. METHODS: From the November 1993 to July 1995 claims data of the University of Connecticut Health Center, asthmatic patient sex, age, racial/ethnic group, address, and health insurance status were examined to identify predictors of treatment site. Patient addresses generated maps and census data. RESULTS: 3288 visits were made by 1455 patients; 8%, 34%, and 58% came from poverty level, low, and higher income residential areas, respectively. Insurance type and then age were the most significant predictors of treatment site. Adults having commercial insurance or Medicare were most likely treated as outpatients, self-pay patients 5 times more likely in the emergency department, and those receiving public assistance 2.4 times more likely in the hospital. Only 9% of Medicaid children and 22% with commercial insurance were evaluated as outpatients. Neither sex nor race/ethnicity was an important predictor of treatment site. CONCLUSION: Although not population-based, this group of asthmatic patients represents a group diverse in socioeconomic status and racial/ethnic background. Insurance category was the most influential factor predicting asthma treatment site, suggesting that economic status may be the most important determinant of higher morbidity. Children were treated predominantly in acute care settings. PMID- 9357384 TI - Serum eosinophil cationic protein levels and bronchodilator responses at acute asthma exacerbation. AB - BACKGROUND: Serum levels of eosinophil cationic protein are an indirect measure of airway inflammation in asthma. It is proposed that the extent to which broncho constriction or airway inflammation contributes to airflow obstruction in acute asthma may determine responsiveness to bronchodilator therapy. OBJECTIVE: To test the hypothesis that subjects with acute asthma exacerbations who respond poorly to inhaled bronchodilator treatment may have more marked airway inflammation than those who respond well to identical therapy. METHODS: Forty-eight asthmatic children who visited the emergency room due to acute exacerbations were studied. Serum levels of eosinophil cationic protein were measured at the time of acute exacerbations and of clinical remissions. At acute exacerbation, FEV1 was assessed before and after the administration of aerosolized salbutamol. RESULTS: The mean serum level of eosinophil cationic protein at acute exacerbation (41.1 +/- 12.8 micrograms/L) was significantly higher (P < .01) than that at clinical remission (30.0 +/- 8.5 micrograms/L) in the study population. The level at acute exacerbation was even higher in group A (n = 18: postbronchodilator FEV1 < 75% predicted) than in group B (n = 30: postbronchodilator FEV1 > or = 75% predicted), whereas both groups showed similar levels at clinical remission. The level at acute exacerbation correlated positively with severity of exacerbation (r = .47, P < .01) and negatively with bronchodilator responses (r = -.56, P < .01). This negative correlation was valid among subjects with a similar degree of exacerbation. CONCLUSION: A higher level of eosinophil cationic protein at acute asthma exacerbation was associated not only with more severe exacerbation but also with a lower degree of bronchodilator responsiveness. This suggests that degree of airway inflammation may be one determinant of degree of responsiveness to initial bronchodilator therapy at acute asthma exacerbation. PMID- 9357385 TI - Comparison of once daily mometasone furoate (Nasonex) and fluticasone propionate aqueous nasal sprays for the treatment of perennial rhinitis. 194-079 Study Group. AB - BACKGROUND: Mometasone furoate (Nasonex), in a new once-daily aqueous nasal spray formulation, has been shown to be as effective and well-tolerated as twice-daily beclomethasone dipropionate aqueous nasal spray in treating symptoms of seasonal allergic rhinitis and perennial rhinitis. OBJECTIVE: To compare the effectiveness and tolerability of mometasone furoate to placebo and of fluticasone propionate aqueous nasal spray, all treatments administered once-daily, in patients with perennial rhinitis. METHODS: This was a 3-month, randomized, double-blind, double dummy, parallel group study in 550 patients, aged 12 to 77 years, at 25 centers in Canada, Latin America, and Europe. Patients allergic to at least one perennial allergen, with confirmed allergy history, skin test positivity, and moderate to severe symptomatology, were eligible to receive one of the following treatments, once daily in the morning: mometasone furoate 200 micrograms, fluticasone propionate 200 micrograms, or placebo. The primary efficacy variable was the change from baseline in total AM plus PM diary nasal symptom score over the first 15 days of treatment. RESULTS: Four hundred fifty-nine patients were valid for efficacy. For the primary efficacy variable, mometasone furoate was significantly (P < .01) more effective than placebo and was not statistically different from fluticasone propionate (percent reductions from baseline were 37, 39, and 22 for mometasone furoate, fluticasone propionate, and placebo, respectively). Generally, similar trends were seen for physician-evaluated total nasal symptoms, and patient-rated and physician-rated overall condition and response to therapy. Overall, mometasone furoate was at least as effective as fluticasone propionate at equivalent doses. There was no evidence of tachyphylaxis. All treatments were well tolerated. CONCLUSION: Mometasone furoate and fluticasone propionate adequately controlled symptoms of perennial rhinitis and were well tolerated. PMID- 9357386 TI - Geriatric clinical pharmacologist and medical oncologist: a new partnership? PMID- 9357387 TI - Validation of the five-drug "Pittsburgh cocktail" approach for assessment of selective regulation of drug-metabolizing enzymes. AB - OBJECTIVES: To determine whether the probe drugs caffeine, chlorzoxazone, dapsone, debrisoquin (INN, debrisoquine), and mephenytoin can be simultaneously administered as a metabolic cocktail to estimate in vivo cytochrome P450 (CYP) and N-acetyltransferase enzyme activities. METHODS: Fourteen healthy nonsmoking male volunteers (mean age +/- SD, 21.6 +/- 2.2 years) received 100 mg caffeine, 250 mg chlorzoxazone, 100 mg dapsone, 10 mg debrisoquin, and 100 mg mephenytoin individually and in four and five-drug combinations in a randomized manner using a 7 x 7 Latin square. Each drug or drug combination was given orally after an overnight fast, with a minimum 1-week washout between administrations. In each session, urine was collected from 0 to 8 hours and plasma was obtained at 4 and 8 hours after drug administration. Plasma and metabolite concentrations were used to estimate phenotypic trait measures for the efficiency of each drug's metabolism. RESULTS: The phenotypic indexes determined for caffeine, chlorzoxazone, dapsone, debrisoquin, and mephenytoin were not significantly different when given alone than when given in combination. The median percentage change of the trait measures observed during administration of all five compounds compared with individual administration ranged from -10.7% for the 6 hydroxychlorzoxazone to chlorzoxazone plasma ratio to +2.2% for the debrisoquin recovery ratio. CONCLUSIONS: The results of this study show that caffeine, chlorzoxazone, dapsone, debrisoquin, and mephenytoin in low doses can be simultaneously administered without metabolic interaction. This cocktail approach can thus simultaneously provide independent in vivo phenotypic measures for multiple CYP enzymes and N-acetyltransferase. PMID- 9357389 TI - Inhibition of the sulfoxidation of omeprazole by ketoconazole in poor and extensive metabolizers of S-mephenytoin. AB - BACKGROUND: The metabolism of omeprazole includes hydroxylation catalyzed by CYP2C19 and, to a minor extent, sulfoxidation, presumably by CYP3A4. Sulfoxidation may be the predominant pathway in individuals devoid of the genetically determined CYP2C19 activity. Ketoconazole is a known CYP3A4 inhibitor in daily doses from 200 to 400 mg. In this study ketoconazole was used as a probe to investigate the extent to which CYP3A4 is involved in omeprazole metabolism in vivo. METHODS: A single oral 20 mg dose of omeprazole before and after four daily doses of 200, 100, or 50 mg ketoconazole was given to 10 healthy subjects, previously phenotyped as poor or extensive metabolizers of S-mephenytoin. Concentrations of omeprazole, 5-hydroxyomeprazole, omeprazole sulfone, and ketoconazole were analyzed with reversed-phase HPLC methods in plasma samples collected repeatedly for 12 hours after dosing. RESULTS: After intake of 20 mg omeprazole with 0, 50, 100, and 200 mg ketoconazole, mean values for omeprazole sulfone area under the plasma concentration versus time curve from 0 to 6 hours [AUC(0-6)] were 482, 206, 167, and < 100 nmol/L.hr in extensive metabolizers and 3160, 2430, 937, and 534 nmol/L.hr in poor metabolizers, respectively. Mean omeprazole AUC(0-6) increased from 1660 to 2265 nmol/L.hr in extensive metabolizers and from 7715 to 15319 nmol/L.hr in poor metabolizers after intake of 200 mg ketoconazole. CONCLUSIONS: An oral daily dose of 100 to 200 mg ketoconazole is sufficient to provide a marked inhibition of the formation of the omeprazole sulfone in both extensive and poor metabolizers and leads to a doubling of omeprazole levels in poor metabolizers, whereas 50 mg ketoconazole provides only partial inhibition. We concluded that CYP3A4 catalyzes the sulfoxidation of omeprazole and that this is the predominant metabolic pathway of omeprazole in poor metabolizers of S-mephenytoin. PMID- 9357388 TI - Isoniazid acetylation metabolic ratio during maturation in children. AB - Isoniazid acetylation metabolic ratio (MR) was studied in 61 children with tuberculosis after administration of isoniazid. MR was calculated as the molar acetylisoniazid to isoniazid concentration ratio. MR was used as a probe for N acetyltransferase activity and to determine the acetylation phenotype. MR had a bimodal distribution with an antimode between 0.48 and 0.77. MR and the percentage of fast acetylators increased significantly with age. The cumulative frequency of fast acetylators increased with age, with a plateau reached around 4 years. MR value was checked during treatment in 44 children. All children but one who initially appeared as fast acetylators remained in this group after repeated testing. Among the 30 slow acetylators, 12 became fast acetylators, and 10 showed a variable phenotyping at different ages. A bimodal distribution of the isoniazid acetylation MR was shown in children, with an antimode close to that described in the literature and a maturation of isoniazid acetylation during the first 4 years. PMID- 9357390 TI - Tobramycin population pharmacokinetics in neonates. AB - OBJECTIVE: To establish a tobramycin dosing schedule for neonates of various gestational ages. METHODS: This was a retrospective study with prospective validation. A retrospective study in 470 neonates, with suspected septicemia in the first week of life, was performed. All patients received tobramycin according to the following scheme: neonates with a gestational age of less than 28 weeks received 3.5 mg/kg every 24 hours, neonates from 28 to 36 weeks received 2.5 mg/kg every 18 hours, neonates older than 36 weeks received 2.5 mg/kg every 12 hours. Trough and peak tobramycin serum levels were determined before drug administration and 30 minutes after the fourth dose. Tobramycin data were analyzed according to a one-compartment open model with use of NONMEM population pharmacokinetic software. Individual empirical Bayes estimates were generated on the basis of the population estimates and used to calculate predicted peak and trough levels for different doses and dosing intervals. To establish an optimal dosing regimen, target trough levels were set at below 2 mg/L and target peak levels were set above 5 to 10 mg/L. The dosing regimen was prospectively evaluated in 23 patients. RESULTS: Of the 470 patients, 19.1% of measured peak and 32.8% of measured trough tobramycin serum levels were outside the desired therapeutic range, and 48.8% of neonates with a gestational age of less than 28 weeks had an aberrant trough level. With use of population estimates, the following dosing regimen was recommended: gestational age below 32 weeks, 4 mg/kg every 48 hours; gestational age between 32 and 37 weeks, 4 mg/kg every 36 hours, gestational age above 37 weeks, 4 mg/kg every 24 hours. With this dosing schedule, predicted peak levels were higher than 5 mg/L in 95.1% of the neonates. Predicted trough levels were higher than 2 mg/L in 1.9% of the neonates and higher than 1 mg/L in 7.6%. Prospectively measured peak levels were higher than 5 mg/L in all but one infant. Measured trough levels were higher than 2 mg/L in three patients and marginally higher than 1 mg/L in four patients. CONCLUSIONS: With the use of this proposed schedule, taking into account differences in gestational ages, predicted peak levels will be therapeutic, whereas predicted trough levels will minimize toxicity. PMID- 9357391 TI - Induction of CYP2D6 in pregnancy. AB - Expression of the drug-metabolizing enzyme cytochrome P4502D6 (CYP2D6) is predominantly under genetic control, and enzyme-inducing drugs have little influence on its activity. We studied the activity of CYP2D6 during pregnancy. One hundred forty pregnant women were genotyped for CYP2D6. Seventeen of them (four poor metabolizers, seven heterozygous extensive metabolizers, and six extensive metabolizers) were phenotyped with dextromethorphan in late pregnancy and 7 to 11 weeks after parturition. During pregnancy the dextromethorphan/dextrorphan metabolic ratio was significantly reduced (p = 0.0015) among homozygous and heterozygous extensive metabolizers, indicating increased CYP2D6 activity. In contrast, poor metabolizers showed an increased metabolic ratio during pregnancy. These results are consistent with previous findings of a marked increase in metabolism of the CYP2D6 substrate metoprolol during pregnancy. Both studies indicate an increase in CYP2D6 activity during pregnancy, which may be caused by an induction of the CYP2D6 enzyme. PMID- 9357392 TI - Effects of age, gender, and diurnal variation on the steady-state pharmacokinetics of BMS-181101, an antidepressant, in healthy subjects. AB - OBJECTIVES: To investigate the effects of age, gender, and diurnal variation on the safety, tolerability, and steady-state pharmacokinetics of BMS-181101, an antidepressant, in humans. METHODS: This was a multiple-dose parallel-design study in 51 healthy subjects (12 young and 12 elderly men and 12 young and 15 elderly women). Each subject received a 15 mg oral dose of BMS-181101 every 12 hours on days 1 through 6 and one dose on day 7. After the evening dose on day 6 and morning dose on day 7, serial blood samples were collected at specified times after administration. Plasma was analyzed for BMS-181101 with use of an HPLC method. RESULTS: Male subjects tolerated BMS-181101 better than female subjects. The mean values for area under the plasma concentration-time curve over the dosing interval tau (AUC tau; 58.8 to 102.4 ng.hr/ml) and elimination half-life t1/2; 5.7 to 10.4 hours) for the elderly subjects were significantly greater than those for the young subjects (39.0 to 64.3 ng.hr/ml and 3.2 to 4.5 hours). The mean values for peak plasma concentration (Cmax; 14.7 to 25.2 ng/ml) and AUC tau (52.4 to 102.4 ng.hr/ml) for the women were significantly greater than those for the men (9.08 to 15.3 ng/ml and 39.0 to 73.6 ng.hr/ml). The mean values for Cmax (14.7 to 25.2 ng/ml) and AUC tau (54.8 to 102.4 ng.hr/ml) on the morning of day 7 were significantly greater than those after the evening dose on day 6 (9.08 to 17.3 ng/ml; 39.0 to 83.4 ng.hr/ml). CONCLUSIONS: An initial lower dose or appropriate titration of daily doses of BMS-181101 may be necessary for the treatment of elderly and female subjects, and the pharmacokinetics of BMS-181101 exhibited significant diurnal effects. PMID- 9357393 TI - Effect of fluconazole on the pharmacokinetics of eprosartan and losartan in healthy male volunteers. AB - OBJECTIVE: To investigate the effect of steady-state fluconazole administration on the disposition of eprosartan, losartan, and E-3174. METHODS: Sixteen healthy male subjects received 300 mg eprosartan every 12 hours, and 16 received 100 mg losartan every 24 hours on study days 1 to 20. All 32 subjects received 200 mg fluconazole every 24 hours beginning on day 11 and continuing through day 20. Serial blood samples were collected over one dosing interval on study days 10 and 20 for measurement of plasma concentrations of eprosartan, losartan, and E-3174 (the active metabolite of losartan). RESULTS: There was no significant difference in eprosartan area under the concentration-time curve from time 0 to time of last quantifiable concentration [AUC(0-t)] or maximum concentration (Cmax) when administered alone and with fluconazole. After concomitant administration with fluconazole, losartan AUC(0-t) and Cmax were significantly increased 66% and 30%, respectively, compared with those values for losartan alone. The AUC(0-t) and Cmax for E-3174 were significantly decreased 43% and 56%, respectively, after administration of losartan with fluconazole. CONCLUSIONS: Fluconazole significantly increases the steady-state AUC of losartan and inhibits the formation of the active metabolite of losartan, E-3174. In contrast, fluconazole administration has no effect on the steady-state pharmacokinetics of eprosartan. PMID- 9357394 TI - Population analysis of the pharmacokinetics and pharmacodynamics of seratrodast in patients with mild to moderate asthma. AB - BACKGROUND: Seratrodast, a potent thromboxane receptor antagonist, is approved in Japan for the treatment of asthma and currently is being developed in the United States. METHODS: This was a phase II, randomized, double-blind, parallel-group, placebo-controlled 15-center study of seratrodast in patients with mild to moderate asthma. A total of 183 patients were randomly assigned to receive daily doses of either placebo, or 80 mg seratrodast, or 120 mg seratrodast for 8 weeks. Pharmacokinetic and pharmacodynamic modeling was carried out by means of the population approach. A two-compartment model with zero-order input and first order elimination best fitted the plasma concentration-time data. RESULTS AND CONCLUSIONS: The pharmacokinetics of seratrodast were linear after single and multiple dosing for 8 weeks. The population estimates for oral clearance and apparent volume of distribution were 8.5 ml/hr/kg and 43.3 ml/kg, respectively. All pharmacokinetic parameters (the oral central compartment clearance, the volumes of distribution of the central and peripheral compartments, and the intercompartmental clearance) were estimated with a precision of 10% or less and were found to be associated with body weight. The residual variability was 30%. The values of oral clearance estimated in this study in male patients were similar to those previously estimated in healthy male subjects. Seratrodast at a dose of 120 mg daily produced an increase in forced expiratory volume in 1 second (FEV1) from baseline that was linearly correlated with its plasma concentrations. The average slope of the concentration-effect relationship was 0.222% and 0.470% per microgram/ml after single and multiple dosing, respectively. Interpatient variability in response was mainly affected by the initial severity of the disease. A lower percentage of predicted FEV1 (i.e., more severe obstruction) was associated with higher slopes, and greater increases in FEV1. PMID- 9357395 TI - Vasodilation in black Americans: attenuated nitric oxide-mediated responses. AB - BACKGROUND: Attenuated vasodilation in response to the intra-arterial administration of the beta-adrenergic agonist isoproterenol (INN, isoprenaline), an endothelium-independent vasodilator, has previously been observed in normotensive black Americans. To determine whether this reflected a more generalized attenuation of responses to vasodilators, we compared forearm blood flow responses to the endothelium-dependent vasodilator methacholine and the endothelium-independent vasodilator sodium nitroprusside in young normotensive black men and white men. METHODS: Forearm blood flow responses to the intra arterial administration of isoproterenol (10 to 400 ng/min), methacholine (0.25 to 8 micrograms/min), and sodium nitroprusside (0.25 to 8 micrograms/min) were measured with use of venous occlusion plethysmography in 11 normotensive black men (mean +/- SE age, 30.5 +/- 2.2 years) and nine normotensive white men (mean age, 28.0 +/- 3.2 years). RESULTS: Baseline characteristics, including baseline forearm blood flow, were similar in the black and the white subjects. Vasodilation in response to isoproterenol, sodium nitroprusside, and methacholine was significantly attenuated in black subjects, resulting respectively in a 3.7 fold, 3.6-fold, and 5.0-fold increase in forearm blood flow in black subjects and a 7.5-fold, 5.2-fold, and 6.9-fold increase in forearm blood flow in white subjects (ANOVA; isoproterenol, p < 0.0001; sodium nitroprusside, p < 0.0001; methacholine, p = 0.01). CONCLUSIONS: Our finding of attenuated nitric oxide mediated vasodilation in response to methacholine and sodium nitroprusside in healthy black American men suggests that attenuated vasodilation in black subjects is a relatively generalized phenomenon, resulting in attenuated responses to multiple vasodilators that act through different receptor- and nonreceptor-mediated mechanisms. PMID- 9357396 TI - Pharmacodynamics of temazepam in primary insomnia: assessment of the value of quantitative electroencephalography and saccadic eye movements in predicting improvement of sleep. AB - BACKGROUND AND OBJECTIVE: Quantitative electroencephalographic parameters and saccadic eye movements are frequently used as pharmacodynamic measures of benzodiazepine effect. We investigated the relationship between these measures and the hypnotic effect. METHODS: The correlation between the pharmacodynamic measures and sleep quality was determined in 21 patients with primary insomnia. The pharmacokinetic-pharmacodynamic relationships were characterized after administration of 20 mg oral temazepam. The hypnotic effect was determined on the basis of polysomnographic sleep recordings and a subjective sleep evaluation questionnaire. Correlations between pharmacodynamic measures and the improvement of sleep were investigated. RESULTS: The pharmacokinetic-pharmacodynamic relationships for the parameters derived from electroencephalography and saccadic eye movements showed considerable interindividual variability. Administration of temazepam led to a significant improvement in the objective parameters sleep period efficiency, wake time after sleep onset, and sleep efficiency and in the subjective assessment of sleep quality. No significant correlations were observed between the pharmacokinetic-pharmacodynamic-derived parameters and the improvement in objective or subjective sleep parameters. CONCLUSION: In subjects with primary insomnia the administration of 20 mg oral temazepam results in changes in both the pharmacodynamic measures and in quality of sleep. No individual correlations between the pharmacodynamic measures and quality of sleep were observed. We concluded that the investigated pharmacodynamic measures are of value in the first assessment of clinical efficacy and for the selection of the dose(s) to be investigated in subsequent trials that aim at showing clinical efficacy. However, the conclusive quantification of clinical efficacy should be performed only on the basis of the clinical end point itself. PMID- 9357397 TI - Arteriovenous differences in plasma concentration of nicotine and catecholamines and related cardiovascular effects after smoking, nicotine nasal spray, and intravenous nicotine. AB - BACKGROUND AND OBJECTIVES: Delivery of a high concentration bolus of nicotine through the arterial circulation is believed to be an important determinant of the addictive, behavioral, and physiologic effects of nicotine. To better understand the pharmacologic features of nicotine with different routes of administration, we measured arterial and venous plasma concentrations of nicotine, cotinine, epinephrine, and norepinephrine after tobacco smoking, intravenous nicotine infusion, and use of a nicotine nasal spray. SUBJECTS AND METHODS: Arterial and venous blood samples were drawn simultaneously from 12 male smokers. Six subjects received a single dose of 1 mg nicotine nasal spray, and six subjects smoked cigarettes, one puff per minute for 10 minutes. All 12 subjects were administered nicotine as a 30-minute infusion beginning 70 minutes after administration of the nicotine nasal spray or commencement of smoking. RESULTS: The mean peak arterial plasma concentrations of nicotine (Cmax) after smoking or administration of nicotine nasal spray, or intravenous nicotine averaged twofold those of venous plasma. For nicotine nasal spray, the time to Cmax was much faster for arterial than for venous plasma (median, 5 versus 18 minutes, p < 0.01). Intravenous nicotine produced the greatest increase in plasma epinephrine concentration, although smoking had a greater chronotropic effect. Acute tolerance to the chronotropic effects of nicotine was suggested at pharmacodynamic analysis with venous nicotine concentrations, whereas analysis of arterial concentrations found the opposite--a time lag between plasma concentration and effect. CONCLUSION: Nicotine is rapidly absorbed from nicotine nasal spray. The Cmax of nicotine after smoking or administration of nicotine nasal spray, or intravenous nicotine is substantially higher in arterial than venous plasma. Acute tolerance to the chronotropic effects of nicotine is not apparent if arterial plasma concentrations are measured. PMID- 9357399 TI - Reducing disability among the elderly in Europe. PMID- 9357398 TI - Olsalazine and 6-mercaptopurine-related bone marrow suppression: a possible drug drug interaction. AB - A patient with refractory Crohn's disease had two separate episodes of bone marrow suppression while receiving 50 to 75 mg 6-mercaptopurine a day and 1000 to 1750 mg olsalazine a day. This adverse reaction necessitated dose reduction of 6 mercaptopurine on the first occasion and withdrawal of 6-mercaptopurine and olsalazine on the second occasion. The patient's red blood cell thiopurine methyltransferase (TPMT) activity was 1.2 U per milliliter red blood cells (low normal range) and her TPMT genotype was wild-type sequence for all known alleles of TPMT that result in low TPMT enzyme activity. In vitro enzyme kinetic studies confirmed the hypothesis that olsalazine and olsalazine-O-sulfate are potent noncompetitive inhibitors of recombinant human TPMT. We suggest that the patient's relatively low baseline level of TPMT activity was inhibited by olsalazine and olsalazine-O-sulfate, leading to decreased clearance of 6 mercaptopurine and its accumulation. This ultimately increased intracellular 6 thiopurine nucleotide levels to toxic concentrations, which caused bone marrow suppression. PMID- 9357400 TI - Cultivation of Whipple bacillus: the irony and the ecstasy. PMID- 9357401 TI - Genetic determination of bone density. PMID- 9357402 TI - Thin cutaneous malignant melanoma and the MIN terminology. PMID- 9357403 TI - Coronary artery calcification on computed tomography. PMID- 9357404 TI - Integrated management of childhood illness. PMID- 9357405 TI - Randomised controlled trial of ketanserin and aspirin in prevention of pre eclampsia. AB - BACKGROUND: Pre-eclampsia is associated with extensive endothelial-cell damage and platelet activation, resulting in lower production of vasodilator prostaglandins and increased release of the vasoconstrictors thromboxane A2 and serotonin. Damage to endothelial-cell serotonin-1 receptors leaves vasoconstriction and platelet aggregation mediated by serotonin-2 receptors unopposed. We investigated the role of ketanserin, a selective serotonin-2 receptor antagonist, in lowering the rate of pre-eclampsia among pregnant women with mild to moderate hypertension. METHODS: We recruited 138 pregnant women into a double-blind, randomised, placebo-controlled trial. They had diastolic blood pressure persistently more than 80 mm Hg before 20 weeks' gestation. 69 women received ketanserin and 69 received placebo. Both groups also received aspirin. Patients were initially given two tablets daily, increased to four tablets daily in diastolic blood pressure was more than 90 mm Hg. Primary outcomes were the development of pre-eclampsia and severe hypertension, and perinatal mortality. FINDINGS: There were significantly fewer cases of pre-eclampsia (two vs 13; relative risk 0.15 [95% CI 0.04-0.66], p = 0.006) and severe hypertension (six vs 17; p = 0.02) in the ketanserin than in the placebo group. There was also a trend towards less perinatal mortality (one vs six deaths) but this was not significant (p = 0.28). Rates of abruptio placentae and pre-eclampsia before 34 weeks' gestation were lower in the ketanserin group, and mean birthweight was significantly higher. INTERPRETATION: We found an association between the addition of ketanserin to aspirin and a decrease in the number of cases of pre eclampsia and severe hypertension, as well as improved pregnancy outcome among patients with mild to moderate midtrimester hypertension. PMID- 9357406 TI - Randomised trial of magnesium in in-hospital cardiac arrest. Duke Internal Medicine Housestaff. AB - BACKGROUND: The apparent benefit of magnesium in acute myocardial infarction, and the persistently poor outcome after cardiac arrest, have led to use of magnesium in cardiopulmonary resuscitation. Because few data on its use in cardiac arrest were available, we undertook a randomised placebo-controlled trial (MAGIC trial). METHODS: Patients treated for cardiac arrest by the Duke Hospital code team were randomly assigned intravenous magnesium (2 g [8 mmoles] bolus, followed by 8 g [32 mmoles] over 24 h; 76 patients) or placebo (80 patients). Only patients in intensive care or general wards were eligible; those whose cardiac arrest occurred in emergency, operating, or recovery rooms were excluded. The primary endpoint was return of spontaneous circulation, defined as attainment of any measurable blood pressure or palpable pulse for at least 1 h after cardiac arrest. The secondary endpoints were survival to 24 h, survival to hospital discharge, and neurological outcome. Analysis was by intention to treat. FINDINGS: There were no significant differences between the magnesium and placebo groups in the proportion with return of spontaneous circulation (41 [54%] vs 48 [60%], p = 0.44), survival to 24 h (33 [43%] vs 40 [50%], p = 0.41), survival to hospital discharge (16 [21%] vs 17 [21%], p = 0.98), or Glasgow coma score (median 15 in both). INTERPRETATION: Empirical magnesium supplementation did not improve the rate of successful resuscitation, survival to 24 h, or survival to hospital discharge overall or in any subpopulation of patients with in-hospital cardiac arrest. PMID- 9357407 TI - Hormone replacement therapy and risk of non-fatal stroke. AB - BACKGROUND: The effect of postmenopausal hormone replacement therapy (HRT) on the risk of subtypes of stroke is as yet unclear. To investigate the effect of oestrogen and combined oestrogen-progestagen therapy on the risk of non-fatal haemorrhagic and thromboembolic stroke, we carried out a case-control study. METHODS: From the Danish National Patient Register we identified all Danish women aged 45-64 years who had a non-fatal, first-ever cerebrovascular attack during 1990-92. Two age-matched controls were randomly selected for each case from the Danish National Person Register. Important correlates of hormone use and stroke, on which information was obtained from postal questionnaires, were controlled for by multivariate analyses based on log-linear graphical models. The analyses included data on 1422 cases classified in four subtypes of stroke (160 subarachnoid haemorrhage, 95 intracerebral haemorrhage, 846 thromboembolic infarction, 321 transient ischaemic attack) and 3171 controls. FINDINGS: After adjustment for confounding variables and correction for the trend in sales of HRT preparations, no significant associations were detected between current use of unopposed oestrogen replacement therapy and non-fatal subarachnoid haemorrhage (odds ratio 0.52 [95% CI 0.23-1.22]), intracerebral haemorrhage (0.15 [0.02 1.09]), or thromboembolic infarction (1.16 [0.86-1.58]), respectively, compared with never use. Current use of combined oestrogen-progestagen replacement therapy had no significant influence on the risk of subarachnoid haemorrhage (1.22 [0.79 1.89]), intracerebral haemorrhage (1.17 [0.64-2.13]), or thromboembolic infarction (1.17 [0.92-1.47]). A significantly increased incidence of transient ischaemic attacks among former users of HRT and among current users of unopposed oestrogen may to some extent be explained by selection--HRT users being more aware of symptoms than non-users. INTERPRETATION: Unopposed oestrogen and combined oestrogen-progestagen replacement therapy have no influence on the risk of non-fatal thromboembolic or haemorrhagic stroke in women aged 45-64 years. PMID- 9357408 TI - Tuberculosis in a prison population in Malawi. AB - BACKGROUND: Much concern has been expressed about the high prevalence of tuberculosis in prisons in industrialised countries. Since there is almost no information from developing countries, we investigated the rate of pulmonary tuberculosis in a large prison in Malawi. METHODS: Between May and July, 1996; we carried out an active case-finding survey in Zomba Central Prison, Malawi, through the National Tuberculosis Control Programme. We interviewed prisoners, and those with a cough of at least 1 week's duration were screened by sputum smear microscopy. If microscopy was negative, prisoners underwent chest radiography. We offered HIV testing, with voluntary consent and counselling before and after tests, to all prisoners, whether positive or negative for pulmonary tuberculosis. FINDINGS: 914 (70%) of 1315 prisoners were screened (905 men, nine women; mean age 30 years [SD 11]). 47 (5%) screened prisoners (all men) had pulmonary tuberculosis: 14 were taking antituberculosis treatment and 33 were undiagnosed at the start of the study; 18 were sputum-smear positive and 15 were sputum-smear negative. 16 (73%) of 22 prisoners with previously undiagnosed pulmonary tuberculosis and 30 (75%) of 40 prisoners with cough but no evidence of pulmonary tuberculosis were HIV seropositive. In all prisoners, except one, symptoms of pulmonary tuberculosis had developed after they had entered prison. INTERPRETATION: We found a high rate of pulmonary tuberculosis in Zomba Central Prison, which suggests active transmission of tuberculosis. As a result of this study, the National Tuberculosis Control Programme has implemented interventions in eight prisons in Malawi to improve tuberculosis control, including collection of health data, education of prisoners and clinical staff about tuberculosis, active screening of prisoners for pulmonary tuberculosis by sputum-smear microscopy, and active case-finding in the prisons. PMID- 9357409 TI - UKPDS 25: autoantibodies to islet-cell cytoplasm and glutamic acid decarboxylase for prediction of insulin requirement in type 2 diabetes. UK Prospective Diabetes Study Group. AB - BACKGROUND: Autoantibodies to islet-cell cytoplasm (ICA) and glutamic acid decarboxylase (GADA) can occur in apparently typical, non-insulin dependent diabetes mellitus (type 2). We investigated whether the presence of either or both antibodies characterises a subtype of diabetes and provides better prediction of requirement for insulin therapy by 6 years' follow-up than clinical variables. METHODS: We measured ICA and GADA at diagnosis of diabetes in a representative population of 3672 white patients with type 2 diabetes, aged between 25 and 65 years. The phenotype was assessed by age of onset, body-mass index, percentage haemoglobin A1c (HbA1c), and islet beta-cell function. We investigated the need for insulin therapy among 1538 patients not assigned insulin and followed up for 6 years from diagnosis. FINDINGS: The proportion of patients with ICA and GADA decreased with increasing age at diagnosis (from 33 [21%] of 157 patients aged 25-34 [corrected] to 66 [4%] of 1769 aged 55-65 for ICA; from 53 [34%] to 122 [7%] for GADA). Among patients younger than 35 at diagnosis, those with ICA or GADA had lower body-mass index than those without (mean 24.9 [SD 6.0] vs 31.7 [7.3] kg/m2; p < 0.0001 and had higher percentage of HbA1c (9.7 vs 8.7%, p < 0.05). 94% of patients with ICA and 84% of those with GADA required insulin therapy by 6 years, compared with 14% of those without the antibodies (p < 0.0001). Among patients older than 55 at diagnosis, the difference between those with and without antibodies in body-mass index was smaller (27.2 [5.4] vs 28.6 [4.8] kg/m2, p < 0.001); 44% of those with ICA, 34% of those with GADA, and 5% with neither antibody required insulin therapy by 6 years (p < 0.0001). Among patients older than 45 years, body-mass index and HbA1c provided little predictive information for insulin requirement, whereas the positive predictive values of GADA (> or = 60 U/L) alone, or both GADA (> or = 20 U/L) and ICA (> 5 U/L), for insulin therapy were 52% and 68%. INTERPRETATION: Among young adults with type 2 diabetes, the phenotype of those with ICA or GADA antibodies was similar to that of classic juvenile-onset insulin-dependent diabetes, and either phenotype or antibodies predicted insulin requirement. In older adults, the phenotype was closer to that of patients without antibodies and only the presence of antibodies predicted an increased likelihood of insulin requirement. PMID- 9357411 TI - Abnormal renal function as a cause of false-positive biochemical screening for Down's syndrome. PMID- 9357410 TI - Delirium and a subclavian abscess. PMID- 9357412 TI - Induction of interstitial collagenase (MMP-1) by UVA-1 phototherapy in morphea fibroblasts. PMID- 9357413 TI - Parathyroid allotransplantation without immunosuppression. PMID- 9357414 TI - Impairment of bronchoprotection by nitric oxide in severe asthma. PMID- 9357415 TI - High mortality rates among patients with tuberculosis in Bangui, Central African Republic. PMID- 9357417 TI - Cyclic vomiting syndrome and mitochondrial DNA mutations. PMID- 9357416 TI - Autoimmune hepatitis type 2 induced by HCV and persisting after viral clearance. PMID- 9357418 TI - Acute pancreatitis during treatment with amiodarone. PMID- 9357419 TI - Bat bite? PMID- 9357420 TI - Hepatic encephalopathy and ascites. AB - The first abnormality leading to sodium and water retention in cirrhosis is the renal tubular defect that is related to deteriorating liver function and hyperaldosteronism. With progression of liver disease and portal hypertension, renal blood flow declines because of the hepatorenal reflex, and is then maintained by the vasoactive hormonal systems. With increasing peripheral vasodilatation, intrarenal factors for maintenance of renal perfusion cause intense cortical vasoconstriction. The systemic vasoactive factors are predominantly compensatory; any attempts to counteract their action risk circulatory collapse. Future studies should be directed at intrarenal factors. The ideal drug for the treatment of portal hypertension would reduce portal pressure, increase renal blood flow, and produce insignificant changes in arterial pressure. PMID- 9357421 TI - Host immunobiology and vaccine development. AB - As the rules of immunoregulation become clearer, the design of vaccines and adjuvants is becoming more scientific. To understand these rules, the interactions between three kinds of cells need to be grasped. Antigen-presenting cells (APCs) initiate the immunoglobulin cascade. The most important of these are dendritic cells, which must first capture antigen, a process aided by particulate matter, the presence of natural or acquired antibodies, or the capacity to activate complement. Then T cells become activated through conjoint action of processed antigenic peptides and APC surface and secreted molecules. T cells mediate inflammation, develop cytotoxic capacity, and help in antibody formation. Whether cells of type 1 or type 2 predominate influences the direction of both cellular and humoral responses. B cells are then activated, leading to antibody formation and often to better antigen presentation. Both T and B cell memory, embedded in long-lived lymphocyte populations, aid heightened immune reactivity when the antigen is re-encountered. The best vaccines stimulate strong memory. PMID- 9357422 TI - Should organs from patients in permanent vegetative state be used for transplantation? International Forum for Transplant Ethics. PMID- 9357423 TI - Terathanasia, folic acid, and birth defects. PMID- 9357424 TI - Terathanasia, folic acid, and birth defects. PMID- 9357425 TI - Terathanasia, folic acid, and birth defects. PMID- 9357426 TI - Persistent vegetative state. PMID- 9357427 TI - Persistent vegetative state. PMID- 9357428 TI - 5-HT2A receptor gene polymorphisms, anorexia nervosa, and obesity. PMID- 9357429 TI - Chronotherapy with 5-fluorouracil, oxaliplatin, and folinic acid in colorectal cancer. PMID- 9357430 TI - Resistance to methicillin. PMID- 9357431 TI - Resistance to methicillin. PMID- 9357432 TI - Methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. PMID- 9357433 TI - Left-ventricular volume reduction or mitral valve reconstruction? PMID- 9357434 TI - Osteoarthritis of the knee. PMID- 9357435 TI - Osteoarthritis of the knee. PMID- 9357436 TI - Mesalazine preparations. PMID- 9357437 TI - Medical editors' trial amnesty. PMID- 9357438 TI - Tuberculosis control in India: role of private doctors. PMID- 9357439 TI - Ultrasound in developing world. PMID- 9357440 TI - Ultrasound in developing world. PMID- 9357441 TI - Immune tolerance mediated by recombinant proteolipid protein prevents experimental autoimmune encephalomyelitis. AB - Proteolipid protein (PLP), a transmembrane protein expressed only in the central nervous system (CNS), is a candidate target autoantigen for autoimmune-mediated demyelination. We have evaluated the effect of a recombinant form of the PLP protein, delta PLP4, in a murine model of experimental autoimmune encephalomyelitis (EAE). PLP-specific T-cell responses were observed following immunization of SJL/J, PL/J and SWR mice with delta PLP4, demonstrating processing of the protein to several distinct antigenic epitopes. Clinical EAE associated with inflammation and demyelination in the CNS also developed after sensitization of mice with delta PLP4 in adjuvant. Conversely, tolerance to delta PLP4 in adult mice and prevention of PLP peptide 139-151-induced EAE was induced by intravenous injection of soluble delta PLP4. The prevention of disease onset was paralleled by a significant reduction in demyelination and CNS inflammatory cell infiltration and diminished PLP139-151-specific T-cell proliferative responses. These results are consistent with the establishment of peripheral T cell tolerance and reinforce the notion that recombinant myelin antigens and intravenous tolerance induction may prove useful in the modulation of the human demyelinating disease, multiple sclerosis (MS). PMID- 9357443 TI - Relative contributions of peripheral and central sources to levels of IL-1 alpha in the cerebral cortex of mice: assessment with species-specific enzyme immunoassays. AB - The peripheral administration or release of cytokines is associated with central nervous system (CNS) effects that are often due to the actions of cytokines behind the blood-brain barrier (BBB). It is not known whether the majority of cytokine behind the BBB is derived from blood or is released from the CNS in response to peripheral signals. We addressed this question for interleukin-1 alpha (IL-1 alpha) by infusing human IL-1 alpha (humIL-1 alpha) into mice and measuring humIL-1 alpha and murine IL-1 alpha (murIL-1 alpha) in cerebral cortex and serum with specific, sensitive enzyme immunoassays. In adult mice receiving 50 micrograms/kg-24 h of humIL-1 alpha subcutaneously for 48 h, brain and blood samples contained humIL-1 alpha but no murIL-1 alpha. This shows that in our study blood-borne IL-1 alpha did not self-stimulate its release in blood or brain. The presence of humIL-1 alpha in brain could only have originated from blood, where it was administered; the brain/blood ratio of 0.126 ml/g indicates that at steady state, brain levels reach about 12% of blood levels. In neonatal mice, both murIL-1 alpha and humIL-1 alpha were detected in brain and blood after the acute subcutaneous injection of humIL-1 alpha. However, the vast majority of immunoactivity in blood and brain was humIL-1 alpha. These results show that most of the IL-1 alpha appearing in response to circulating IL-1 alpha in areas of the CNS behind the BBB is due to passage across the BBB and not to release from stores endogenous to the CNS. PMID- 9357442 TI - Prolactin is an autocrine growth factor for the Jurkat human T-leukemic cell line. AB - Despite convincing evidence of cooperation between IL-2 and endogenous prolactin (PRL) during T cell activation, the individual role of PRL as a T-cell lineage cytokine remains to be defined. We have examined the production and function of PRL on the Jurkat human T-leukemic cell line, which does not constitutively produce IL-2. The majority of Jurkat cells expressed PRL receptor (R) under standard culture conditions, whereas appearance of the alpha chain of the IL-2-R required PHA-PMA stimulation, as did IL-2 synthesis. Western blotting revealed a predominant band at 23.5 kDa and a weaker band at 25.5 kDa in both Jurkat cell lysates and human (h) pituitary PRL. Metabolic labeling of the cell lysates with 35S-methionine and immunoprecipitation with an antiserum against hPRL showed that both forms of PRL are actively synthesized by the Jurkat cell line. PRL released in the medium was biologically active in the rat Nb2 lymphoma mitogenic assay. Depletion of medium PRL with two polyclonal anti-hPRL antisera inhibited the growth of Jurkat cells in a dose-dependent manner, as evaluated by cell number and 3H-TdR uptake. Purified pituitary or recombinant hPRL at a wide range of concentrations had no significant effect on their growth, but reversed the blocking activity of the anti-hPRL antibody. Recombinant IL-2 had no effect on the antibody-induced growth inhibition. Taken as a whole, these results demonstrate that PRL can act as an autocrine T cell growth factor independently of IL-2 and are the first evidence of its involvement in human leukemic growth and possibly in leukemic transformation. PMID- 9357444 TI - Immune system genes in multiple sclerosis: genetic association and linkage analyses on TCR beta, IGH, IFN-gamma and IL-1ra/IL-1 beta loci. AB - The role of genetic factors in the etiology of multiple sclerosis (MS) has been clearly demonstrated but the loci determining susceptibility to this disease remain largely unidentified. A contribution from several immune system genes has been suggested based on animal models and association/linkage analyses on MS patients and families. With the exception of the findings from the HLA complex, studies on candidate immune system genes have provided controversial results. Here we have performed genetic association and linkage analyses on four chromosomal regions containing immune system genes. A possible role for each of these loci in MS has been previously suggested. In data-sets derived from the Finnish population we found no evidence for contribution of the T-cell receptor beta chain (TCR beta chromosome 7q35), immunoglobulin heavy chain (IGH chromosome 14q32), interferon-gamma (IFN-gamma chromosome 12q14-q15) or interleukin-1 receptor antagonist/interleukin-1 beta (IL-1ra/IL-1 beta chromosome 2q14-q21) loci in the genetic susceptibility to MS. PMID- 9357445 TI - A low virulent strain of Candida albicans enhances brain anticryptococcal defenses: characterization of the local immune reaction by RT-PCR and histochemical analysis. AB - Here we studied the involvement of PCA-2, a low-virulent strain of Candida albicans known to act as a potent stimulating agent in the development of cryptococcal meningoencephalitis. To this purpose, mice received saline or PCA-2 intracerebrally 7 days before lethal local challenge with Cryptococcus neoformans. We found that, following C. neoformans challenge, PCA-2-treated but not saline-treated mice exhibited (a) delayed brain colonization, (b) enhanced median survival times, (c) massive local immune reaction consisting of abundant astrocytes, microglial and inflammatory cells, and (d) a peculiar trend of cytokine gene expression, including high steady-state levels of interleukin (IL) 1 beta and tumor necrosis factor alpha transcripts, fluctuating levels of interferon gamma and inducible nitric oxide synthase mRNA and lately detectable IL-6 gene expression. PCA-2-mediated immunostimulating properties were partially impaired by aminoguanidine or pentoxifylline treatment, further strengthening the conclusion that soluble mediators, including proinflammatory cytokines and nitric oxide, are important defense elements against cryptococcal meningoencephalitis. PMID- 9357446 TI - Linomide activates the adrenocortical axis in the rat: inhibition of experimental autoimmune encephalomyelitis by linomide is not related to the increase of corticosterone. AB - Linomide is a synthetic compound that affects various immunological functions and inhibits experimental autoimmune encephalomyelitis (EAE). In the present study we evaluated the effect of linomide on the HPA axis functions under basal and stress induced conditions and examined whether the effect of linomide on the HPA axis is involved in linomide-induced amelioration of EAE in rats. Linomide caused a significant increase of serum ACTH and corticosterone (CS). The adrenocortical response to various stress modalities as well as the negative feedback exerted by glucocorticoids was not affected. The marked reduction of thymus weight following linomide treatment was abrogated in adrenalectomized rats. The induction of EAE in adrenalectomized rats was completely inhibited by linomide treatment. These results suggest that the increased CS levels induced by linomide are responsible for the decrease in thymus weight but do not play a role in the therapeutic effect of this drug in EAE. PMID- 9357448 TI - Humoral immune response against Campylobacter jejuni lipopolysaccharides in Guillain-Barre and Miller Fisher syndrome. AB - In this study we characterized the IgG antibodies against lipopolysaccharides (LPS) of Campylobacter jejuni in serum from patients with Guillain-Barre syndrome (GBS), Miller Fisher syndrome (MFS), C. jejuni enteritis and normal controls. In patients with GBS and MFS long-lasting titers of IgG1 and IgG3 antibodies against LPS from GBS and MFS associated C. jejuni were found. The subclass and course of these antibodies were highly associated with those of antibodies against GM1 and GQ1b in GBS and MFS patients. However, in C. jejuni enteritis and normal controls anti-LPS antibodies were predominantly IgG2. Antibody binding with LPS was reduced after treatment with choleratoxin and sialidases, suggesting that the ganglioside-like epitopes in LPS are immunodominant. These results further indicate that antecedent C. jejuni infections determine the specificity and isotype of anti-ganglioside antibodies in GBS and MFS patients. PMID- 9357447 TI - Therapeutic potential of phosphodiesterase type 4 inhibition in chronic autoimmune demyelinating disease. AB - It was recently demonstrated that selective phosphodiesterase type 4 (PDE4) inhibition suppresses the clinical manifestations of acute experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), and inhibits the production of tumor necrosis factor-alpha (TNF-alpha), a pathogenetically central cytokine. Since the most common presentation of MS in humans is a relapsing-remitting course, we investigated the therapeutic potential of PDE4 inhibition in the relapsing-remitting EAE model of the SJL mouse. Administration of rolipram, the prototypic PDE4 inhibitor, reduced the clinical signs of EAE during both the initial episode of disease and subsequent relapses. In parallel, there was marked reduction of demyelination and also less inflammation throughout the central nervous system (CNS) of rolipram-treated animals. Gene expression of proinflammatory cytokines in the CNS was reduced in most of the rolipram-treated animals. Additional experiments demonstrated that PDE4 inhibition acted principally by inhibiting the secretion of Th1 cytokines, however, the encephalitogenic potential of myelin basic protein-specific T cells was not impaired. Our findings suggest that PDE4 inhibitors are a promising cytokine-directed therapy in chronic demyelinating disease. PMID- 9357449 TI - T-cell receptor V beta-element expression in peripheral nerves of Lewis rats suffering from experimental autoimmune neuritis. AB - In experimental autoimmune neuritis (EAN), peripheral nerves are infiltrated by T lymphocytes and macrophages. By RT-PCR and sequence analysis we characterized TCR V beta-element usage in sciatic nerve tissue of Lewis rats suffering from EAN induced by immunization with peripheral myelin antigens. Several TCR V beta-chain sequences were detected, which did not show homology to sequences of P2-reactive T cells published so far. In EAN induced with peripheral nerve myelin, but not with P2-protein or P2 peptide aa 53-78, TCR V beta 8.2 sequences identical to sequences of encephalitogenic myelin basic protein (MBP) reactive T-cells were identified. These results provide further evidence for a contribution of MBP directed T-cell reactivity to the pathogenesis of myelin induced EAN and may have implications for the pathogenesis of human demyelinating neuropathies. PMID- 9357451 TI - Neuronal-associated tumor necrosis factor (TNF alpha): its role in noradrenergic functioning and modification of its expression following antidepressant drug administration. AB - Tumor necrosis factor-alpha (TNF alpha) and the alpha 2-adrenergic agonist clonidine regulate norepinephrine (NE) release from noradrenergic nerve terminals in the central nervous system (CNS). In the present study, superfusion and electrical field stimulation were applied to a series of rat hippocampal brain slices in order to investigate the regulation of [3H]-NE release. NE release had been previously determined to be decreased by TNF alpha in a concentration dependent manner, an effect which was potentiated by the alpha 2-adrenergic antagonist idazoxan. Presently, we demonstrate that similar to alpha 2-adrenergic activation, TNF alpha regulation of NE release in a region of the brain rich in noradrenergic nerve terminals, is dependent upon the frequency of electrical stimulation applied to the hippocampal slice. Furthermore, immunoperoxidase staining has verified our previous findings of constitutive TNF alpha protein in the rat brain. Staining for TNF alpha appears to be largely localized to neurons and neuronal processes, further substantiating the proposal that TNF alpha is either synthesized de novo or is accumulated in and released by neurons. After administration of the tricyclic antidepressant desipramine, tissue sections obtained from the rat hippocampus and locus coeruleus are devoid of neuronal associated TNF alpha immunoreactivity. TNF alpha localization in neurons and its modification of NE release comparable to alpha 2-adrenergic receptor activation, explains a functional role for the cytokine as a neuromodulator in the CNS. PMID- 9357450 TI - Effects of beta-IFN-1b treatment in MS patients on adhesion between PBMNCs, HUVECs and MS-HBECs: an in vivo and in vitro study. AB - The in vivo effects on the expression of adhesion molecules and on the adhesion between mononuclear cells and multiple sclerosis human brain endothelial cells (MS-HBECs) were investigated at the beginning of beta-IFN-1b treatment of MS patients. MS-HBECs were isolated from a surgical specimen obtained from an MS patient undergoing brain surgery for vascular aneurysm. 48 h after the first single administration of beta-IFN-1b, PBMNCs of 10 MS patients were analyzed for HLA-DR, CD11a, CD18 and VLA-4 expression and the adhesion between PBMNCs and both stimulated and unstimulated MS-HBECs evaluated. sICAM-1 and sVCAM-1 dosage in the serum of the patients was checked as well. The experiments were repeated using HUVECs in order to detect possible endothelial organ-specific differences. The experiments were also performed after six months of beta-INF-1b treatment on HUVECs. No significant effects on mononuclear cells/endothelium adhesion were detected at 48 h, but adhesion of PBMNCs to HUVECs decreased at six months. An increase in HLA-DR and VLA-4 and a decrease of CD18 was detected in monocytes. The serum level of sVCAM-1 increased at T2 and was still higher than at T0 at six months. The effect of the beta-IFN-1b treatment on both MS-HBECs and HUVECs, was selectively studied in vitro by testing the expression of cytokine-induced adhesion molecules HLA-DR, ICAM-1 and VCAM-1. The in vitro experiments confirmed that beta-IFN-1b is able to antagonize gamma-IFN-induced HLA-DR expression on MS human brain endothelial cells without relevant effects on VCAM-1 and ICAM-1. PMID- 9357452 TI - T helper 2 cytokines induce preproenkephalin mRNA expression and proenkephalin A in human peripheral blood mononuclear cells. AB - We investigated the regulatory influence of several cytokines on the expression of preproenkephalin (PPE) mRNA in human peripheral blood mononuclear cells (PBMC). By use of a quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR), we demonstrate that the T helper 2 cytokines IL-4 and IL-10 are more potent in upregulating PPE mRNA expression in human PBMC than the T helper 1 cytokines IL-2 and gamma-IFN. In addition, TGF-beta is also an effective inducer of PPE mRNA. TGF-beta, IL-4 and IL-10 increase the cytoplasmatic concentration of met-enkephalin in PBMC. Secretion of met-enkephalin in the culture supernatant of IL-4- or IL-10-stimulated PBMC could not be observed, but proenkephalin A-derived met-enkephalin containing peptides could be demonstrated. IL-4 and IL-10 do not induce PPE mRNA via the same pathways. We could observe that PKA is involved in IL-4 mediated PPE mRNA induction, whereas IL-10 apparently uses another route. PMID- 9357454 TI - Is intermittent claudication improved by percutaneous transluminal angioplasty? A randomized controlled trial. AB - PURPOSE: Percutaneous transluminal angioplasty (PTA) is an increasingly popular invasive treatment for peripheral arterial disease, but there have been very few controlled trials to justify its use. This randomized controlled clinical trial was performed to determine in patients with mild and moderate intermittent claudication differences in outcome between PTA and conventional medical treatment after 2 years. METHODS: Six hundred patients with claudication were screened at the Peripheral Vascular Clinic, Royal Infirmary of Edinburgh. Sixty two patients with short femoral artery stenoses or occlusions (47 patients) and iliac stenoses (15 patients) were randomized to either PTA plus medical treatment (PTA group, 30 patients) or to medical treatment alone (control group, 32 patients). Medical treatment consisted of daily low-dose aspirin and advice on smoking and exercise. Outcome measures studied were patient-reported maximum walking distance, exercise treadmill distance until onset of claudication, treadmill maximum walking distance, ankle-brachial pressure index (ABPI), quality of life (Nottingham Health Profile), and duplex ultrasound-measured extent of occlusive disease. RESULTS: At 2 years of follow-up, the PTA group and control subjects did not differ significantly in patient-reported maximum walking, treadmill onset to claudication, treadmill maximum walking distances, or ABPI (p > 0.05). However, the PTA group had significantly fewer occluded arteries (p = 0.003) and a lesser degree of stenosis (expressed in terms of the velocity ratio; p = 0.004) in patent arteries. Quality of life was not demonstrably different between the two groups (p > 0.05). CONCLUSIONS: Two years after PTA, patients had less extensive disease than medically treated patients, but this did not translate into a significant advantage in terms of improved walking or quality of life. There are important implications for patient management and future clinical research. PMID- 9357455 TI - Results of aortic bifurcation grafts for aortoiliac occlusive disease: a meta analysis. AB - PURPOSE: To summarize mortality, morbidity, and long-term patency data of bifurcated aortoiliac or aortofemoral bypass graft procedures in aortoiliac occlusive disease. METHODS: A Medline search was performed of the medical literature published between 1970 and 1996. Studies were included if (1) they reported patency rates based on life tables and the number at risk was provided at yearly intervals; and (2) patient and study characteristics were reported in sufficient detail. Mortality and morbidity risks were pooled using a fixed effects model. The patency data were combined using a technique that enables adjustment for differences across studies in patient characteristics or reporting methods. In the current analysis, we corrected for the symptomatic status of the patients at the time of surgery (claudication vs ischemia) and the unit of observation used in reporting the patency (limb vs patient). RESULTS: We identified 23 studies that met the inclusion criteria. The aggregated operative mortality risk in the older studies (started before 1975) was 4.6%, as compared with 3.3% in the more recent studies (p = 0.01). The aggregated systemic morbidity risk was 13.1% in the older studies and 8.3% in the more recent studies (p < 0.001). Limb-based patency rates for patients with claudication were 91.0% and 86.8% at 5 and 10 years, respectively, as compared with 87.5% and 81.8% for patients with ischemia. Patency rates reported in the older studies were markedly similar to those of more recent studies (p = 0.58). CONCLUSIONS: Our study suggests that mortality and systemic morbidity rates of aortic bifurcation graft procedures have dropped since 1975, whereas patency rates seem to be fairly constant over the years. PMID- 9357453 TI - Randomized trial comparing infrainguinal polytetrafluoroethylene bypass grafting with and without vein interposition cuff at the distal anastomosis. The Joint Vascular Research Group. AB - PURPOSE: A multicenter randomized prospective study was undertaken to determine whether an interposition vein cuff improved the short-term and medium-term patency and limb salvage rates of femoral-above-knee and femoral-below-knee popliteal artery polytetrafluoroethylene (PTFE) bypass procedures. METHODS: Two hundred sixty-one bypass operations were randomized (133 to vein cuff and 128 to no vein cuff). One hundred fifty grafts were to the above-knee popliteal artery, 96 to the below-knee popliteal artery, and 15 to tibial vessels. The median follow-up was 617 days. RESULTS: The 12-month patency rates for cuffed and uncuffed above-knee popliteal artery PTFE bypass grafts were 80% and 84%, and the 2-year patency rates were 72% and 70%, respectively. The patency rates for bypass grafts to the below-knee popliteal artery at 12 months were 80% and 65% and at 2 years 52% and 29%, respectively (p = 0.03). At the below-knee site, this was reflected in 24-month difference in limb salvage rates of 84% and 62%, respectively (p = 0.08). CONCLUSIONS: There was no improvement in the patency rate with the use of a distal anastomosis interposition vein cuff in femoral above-knee popliteal PTFE bypass grafts, but there was a statistically significant advantage when PTFE bypass grafts were anastomosed to the popliteal artery below the knee. PMID- 9357456 TI - Perioperative ischemia and cardiac complications in major vascular surgery: importance of the preoperative twelve-lead electrocardiogram. AB - PURPOSE: To investigate the associations between specific preoperative 12-lead electrocardiogram (ECG) abnormalities, perioperative ischemia, and postoperative myocardial infarction or cardiac death in major vascular surgery. METHODS: Two prospective studies on perioperative myocardial ischemia performed in two tertiary university hospitals were combined to include 405 patients. All preoperative ECGs were analyzed according to the Sokolow-Lyon criteria for left ventricular hypertrophy by investigators who were blinded to the patients' perioperative clinical course. Perioperative myocardial ischemia was detected by continuous ECG recording, and postoperative cardiac complications included myocardial infarction and cardiac death. RESULTS: A total of 19 postoperative cardiac complications occurred (two cardiac deaths and 17 myocardial infarctions). Voltage criteria for left ventricular hypertrophy (78 patients, 19%) and ST segment depression greater than 0.5 mm (98 patients, 24.2%) on preoperative ECGs were both significantly associated with postoperative myocardial infarction or cardiac death (odds ratio, 4.2 and 4.7; p = 0.001 and 0.0005, respectively) and with longer intraoperative and postoperative myocardial ischemia. In each of the two study groups, a preoperative ECG abnormality that involved voltage criteria, ST segment depression, or both (134 patients, 33.1%) was more predictive of postoperative cardiac complications than any other preoperative clinical variable, including a history of myocardial infarction or angina pectoris, diabetes mellitus, pathologic Q-wave by ECG, or preoperative myocardial ischemia. The combined duration of intraoperative and postoperative ischemia and the preoperative ECG with either voltage criteria or ST segment depression were the only independent factors associated with adverse cardiac events by multivariate analysis (p < or = 0.0001 and p = 0.02, respectively). CONCLUSION: Left ventricular hypertrophy and ST segment depression on preoperative 12-lead ECGs are important markers of increased risk for myocardial infarction or cardiac death after major vascular surgery. PMID- 9357457 TI - Prevention of postoperative thrombotic stroke after carotid endarterectomy: the role of transcranial Doppler ultrasound. AB - PURPOSE: To determine the incidence of particulate embolization after carotid endarterectomy (CEA), the effect of dextran-40 infusion in patients with sustained postoperative embolization, and the impact of transcranial Doppler (TCD) monitoring plus adjuvant dextran therapy on the rate of postoperative carotid thrombosis. METHODS: Prospective study in 100 patients who underwent CEA with 6-hour postoperative monitoring using a TCD that was modified to allow automatic, intermittent recording from the ipsilateral middle cerebral artery waveform (10 minute sample every 30 minutes). An incremental dextran-40 infusion was commenced if 25 or more emboli were detected in any 10-minute period. RESULTS: Overall, 48% of patients had one or more emboli detected in the postoperative period, particularly in the first 2 hours. However, sustained embolization that required Dextran therapy developed in only five patients. In each case, the rate of embolization rapidly diminished. CONCLUSIONS: A small proportion of patients have sustained embolization after CEA, which in previous studies has been shown to be highly predictive of thrombotic stroke. Intervention with dextran reduced and subsequently stopped all the emboli in those in whom it was used and contributed to a 0% perioperative morbidity and mortality rate in this series. PMID- 9357458 TI - Juxtalumenal location of plaque necrosis and neoformation in symptomatic carotid stenosis. AB - PURPOSE: The structural features that underlie carotid plaque disruption and symptoms are largely unknown. We have previously shown that the chemical composition and structural complexity of critical carotid stenoses are related to plaque size regardless of symptoms. To further determine whether the spatial distribution of individual plaque components in relation to the lumen corresponds to symptomatic outcome, we evaluated 99 carotid endarterectomy plaques. METHODS: Indications for operation were symptomatic disease in 59 instances (including hemispheric transient ischemic attack in 29, stroke in 19, and amaurosis fugax in 11) and angiographic asymptomatic stenosis > 75% in 40. Plaques removed after remote symptoms beyond 6 months were excluded. Histologic sections from the most stenotic region of the plaque were examined using computer-assisted morphometric analysis. The percent area of plaque cross-section occupied by necrotic lipid core with or without associated plaque hematoma, by calcification, as well as the distance from the lumen or fibrous cap of each of these features, were determined. The presence of foam cells, macrophages, and inflammatory cell collections within, on, or just beneath the fibrous cap was taken as an additional indication of plaque neoformation. RESULTS: The mean percent angiographic stenosis was 82% +/- 11% and 79% +/- 13% for the asymptomatic and symptomatic groups, respectively (p > 0.05). The necrotic core was twice as close to the lumen in symptomatic plaques when compared with asymptomatic plaques (0.27 +/- 0.3 mm vs 0.5 +/- 0.5 mm; p < 0.01). The percent area of necrotic core or calcification was similar for both groups (22% vs 26% and 7% vs 6%, respectively). There was no significant relationship to symptom production of either the distance of calcification from the lumen or of the percent area occupied by the lipid necrotic core or calcification. The number of macrophages infiltrating the region of the fibrous cap was three times greater in the symptomatic plaques compared with the asymptomatic plaques (1114 +/- 1104 vs 385 +/- 622, respectively, p < 0.009). Regions of fibrous cap disruption or ulceration were more commonly observed in the symptomatic plaques than in the asymptomatic plaques (32% vs 20%). None of the demographic or clinical atherosclerosis risk factors distinguished between symptomatic and asymptomatic plaques. CONCLUSIONS: These findings indicate that proximity of plaque necrotic core to the lumen and cellular indicators of plaque neoformation or inflammatory reaction about the fibrous cap are associated with clinical ischemic events. The morphologic complexity of carotid stenoses does not appear to determine symptomatic outcome but rather the topography of individual plaque components in relation to the fibrous cap and the lumen. Imaging techniques that precisely resolve the position of the necrotic core and evidence of inflammatory reactions within carotid plaques should help identify high-risk stenoses before disruption and symptomatic carotid disease. PMID- 9357459 TI - Relationship of age, gender, race, and body size to infrarenal aortic diameter. The Aneurysm Detection and Management (ADAM) Veterans Affairs Cooperative Study Investigators. AB - PURPOSE: To assess the effects of age, gender, race, and body size on infrarenal aortic diameter (IAD) and to determine expected values for IAD on the basis of these factors. METHODS: Veterans aged 50 to 79 years at 15 Department of Veterans Affairs medical centers were invited to undergo ultrasound measurement of IAD and complete a pre-screening questionnaire. We report here on 69,905 subjects who had no previous history of abdominal aortic aneurysm (AAA) and no ultrasound evidence of AAA (defined as IAD > or = 3.0 cm). RESULTS: Although age, gender, black race, height, weight, body mass index, and body surface area were associated with IAD by multivariate linear regression (all p < 0.001), the effects were small. Female sex was associated with a 0.14 cm reduction in IAD and black race with a 0.01 cm increase in IAD. A 0.1 cm change in IAD was associated with large changes in the independent variables: 29 years in age, 19 cm or 40 cm in height, 35 kg in weight, 11 kg/m2 in body mass index, and 0.35 m2 in body surface area. Nearly all height-weight groups were within 0.1 cm of the gender means, and the unadjusted gender means differed by only 0.23 cm. The variation among medical centers had more influence on IAD than did the combination of age, gender, race, and body size. CONCLUSIONS: Age, gender, race, and body size have statistically significant but small effects on IAD. Use of these parameters to define AAA may not offer sufficient advantage over simpler definitions (such as an IAD > or = 3.0 cm) to be warranted. PMID- 9357460 TI - Effect of intraluminal thrombus on abdominal aortic aneurysm wall stress. AB - PURPOSE: Abdominal aortic aneurysms (AAAs) rupture when the wall stress exceeds the strength of the vascular tissue. Intraluminal thrombus may absorb tension and reduce AAA wall stress. This study was performed to test the hypothesis that intraluminal thrombus can significantly reduce AAA wall stress. METHODS: AAA wall stresses were determined by axisymmetric finite element analysis. Model AAAs had external diameters ranging from 2.0 to 4.0 cm. Model parameters included: AAA length, 6 cm; wall thickness, 1.5 mm; Poisson's ratio, 0.49; Young's modulus, 1.0 MPa; and luminal pressure, 1.6 x 10(5) dyne/cm2. Stresses were calculated for each model without thrombus, and then were recalculated with thrombus filling 10% of the AAA cavity. Calculations were repeated as thrombus size was increased in 10% increments and as thrombus elastic modulus increased from 0.01 MPa to 1.0 MPa. Maximum wall stresses were compared between models that had intraluminal thrombus and the unmodified models. Stress reduction greater than 25% was considered significant. RESULTS: The maximum stress reduction of 51% occurred when thrombus with elastic modulus of 1.0 MPa filled the entire AAA cavity. Stresses were reduced by only 25% as modulus decreased to 0.2 MPa. Similarly, decreasing thrombus size by 70% resulted in stress reduction of only 28%. Large AAAs experienced greater stress reduction than small AAAs (48% vs 11%). CONCLUSION: Intraluminal thrombus can significantly reduce AAA wall stress. PMID- 9357462 TI - Observations on delayed neurologic deficit after thoracoabdominal aortic aneurysm repair. AB - PURPOSE: To describe the phenomenon of delayed-onset neurologic deficit after thoraco-abdominal aortic aneurysm repair and to discuss management of this type of deficit in a case series. METHODS: Since September 1992 we have used cerebrospinal fluid drainage and distal aortic perfusion routinely to reduce the risk of neurologic deficit in thoracoabdominal aortic aneurysm patients. All patent intercostal arteries were reattached when this was technically feasible. Delayed neurologic complications occurred in eight patients who underwent operation for thoracoabdominal aortic aneurysm between September 1992 and March 1997, between 1 and 14 days after awakening from anesthesia. All patients had immediate cerebrospinal fluid drainage on recognition of their symptoms. RESULTS: Patients were evaluated by an independent neurologist and were classified by a modified Tarlov score between 0 and 5. All eight patients improved at least two points by discharge. The mean change in Tarlov score from onset to discharge was 2.4 +/- 1.1 (p = 0.008). CONCLUSIONS: Cerebrospinal fluid drainage significantly improved late-onset neurologic deficit that occurred between 1 day and 2 weeks after operation in our series. Immediate drainage should be considered when signs of neurologic deficit first begin to appear. PMID- 9357461 TI - Coagulopathy associated with residual dissection after surgical treatment of type A aortic dissection. AB - PURPOSE: This study was performed to evaluate the effects of a residual dissection on coagulation, fibrinolysis, and platelet function after surgical treatment of acute type A aortic dissection. METHODS: Between 1987 and 1995, 48 consecutive patients underwent emergency surgery for acute type A aortic dissection. Thirty-five of 41 survivors were followed-up for periods ranging from 6 to 112 months (median, 30.3 months). These survivors were classified into three groups by computed tomographic scanning and angiography. Fifteen patients had no residual dissection (group I). Of the 20 patients who had residual dissection, nine had an enlarged aorta greater than 45 mm in maximal diameter (group II), and 11 had an aorta less than 45 mm in maximal diameter (group III). For all patients, blood samples were collected for coagulation, fibrinolysis, and platelet function studies on the same day that the computed tomographic scanning had been performed. RESULTS: beta-thromboglobulin, thrombin-antithrombin III complex, D-dimer, and alpha 2 plasmin inhibitor-plasmin complex concentrations were significantly higher in group II than in the other two groups. Strong correlations between the maximal diameter of the dissected aorta and beta thromboglobulin, thrombin-antithrombin III complex, D-dimer, and plasmin inhibitor-plasmin complex concentrations were evident. In contrast, correlations between the length of the dissected aorta and coagulation/fibrinolysis measurements were weak. CONCLUSIONS: Our findings suggest that the coagulopathy worsened in proportion to the degree of dilatation of the dissected aorta. PMID- 9357463 TI - Measurement of spinal cord blood flow by an inhalation method and intraarterial injection of hydrogen gas. AB - PURPOSE: This study was performed to determine spinal cord blood flow using the H2 clearance method. METHODS: In 12 dogs we measured blood flow determined by hydrogen clearance techniques (BF) in both gray and white matter by spinal cord puncture, and compared the results with BF measured by a catheter inserted intrathecally to avoid spinal cord injury. We studied direct intraarterial and intravenous injection of hydrogen in addition to the inhalation method because hydrogen is an explosive gas. RESULTS: BF measured intrathecally with catheters adherent to either the ventral or the dorsal funiculus did not differ significantly from that of the gray matter. BF measured with a catheter inserted into the epidural space was about one fourth of the BF measured intrathecally. Values measured by the intraarterial injection method did not differ significantly from those obtained by the inhalation method. CONCLUSIONS: BF measured with a catheter inserted intrathecally reflects blood flow in the gray matter of the spinal cord. Using intraarterial injection of H2, BF was safety and accurately measured while avoiding the risk of explosion. PMID- 9357464 TI - Skin perfusion pressure measurement is valuable in the diagnosis of critical limb ischemia. AB - PURPOSE: Critical limb ischemia (CLI) is equated with a need for limb salvage. Arterial reconstruction and major amputation are the therapies ultimately available to such patients. We studied whether measurements of skin perfusion pressure (SPP) can be used to accurately identify those patients with CLI who require vascular reconstruction or major amputation and distinguish them from patients whose foot ulcer would heal with local wound care or minor amputation. METHODS: Fifty-three patients with a total of 61 limbs with a nonhealing foot ulcer (age range, 47 to 88 years; mean, 70.8 +/- 9.8 years; 33 men, 20 women) who were referred to the Vascular Laboratory at Morristown Memorial Hospital for evaluation of arterial insufficiency were studied in a prospective, double blinded fashion. Patients were included in the study if informed consent was obtained, and patients were excluded if there was uncontrolled sepsis or if they required guillotine amputation. The size and site of the foot ulcer was recorded. If gangrene was present, the location and extent was also noted. The pulses were examined and recorded, and the ankle-brachial index was determined for each limb. Measurements of SPP were made at the proximal margin of the ulcer in viable tissue (not in the bed of the ulcer). SPP measurements were made independent of the vascular surgeon's evaluation of the limb and were not part of his clinical decision regarding management of the foot ulcer. The SPP measurements were compared (Fischer's exact test) with the clinical decision for therapy (group I, arterial reconstruction or major amputation; or group II, wound debridement, minor amputation, or both). SPP was also compared with the outcome (ulcer healed or failed to heal) of therapy in group II. From contingency tables we calculated the sensitivity, specificity, positive and negative predictive values (PPV, NPV), and the overall accuracy of SPP measurement as a diagnostic test for critical limb ischemia. RESULTS: There was no difference in the size or location of foot ulcers between groups I and II, nor was there a difference in ulcer size or location between limbs that healed and did not heal in group II. The prevalence of diabetes was similar in all groups and subgroups. The ABI was not predictive of the need for reconstruction or major amputation nor the outcome of local therapy. SPP measurements identified 31 of 32 limbs diagnosed as having CLI by clinical evaluation (i.e., group I, those limbs that required vascular reconstruction or major amputation). Of those patients who were clinically assessed as not having CLI (group II), SPP measurements diagnosed 12 of the 14 limbs that did not heal as having CLI (PPV, 75%) and 11 of 15 limbs that did heal as not having CLI (NPV, 85%). The sensitivity of SPP less than 30 mm Hg as a diagnostic test of CLI was 85%, and the specificity was 73%. The overall diagnostic accuracy of SPP less than 30 mm Hg as a diagnostic test of critical limb ischemia was 79.3% (p < 0.002, Fischer's exact test). CONCLUSIONS: We conclude that SPP measurement is an objective, noninvasive method that can be used to diagnose critical limb ischemia with approximately 80% accuracy. PMID- 9357465 TI - Variability and reliability of air plethysmographic measurements for the evaluation of chronic venous disease. AB - PURPOSE: Air plethysmography (APG) has the potential to help evaluate different treatments for the prevention of recurrence of venous ulcers; however, there are little reported data on the variation and reliability of the different parameters. This study aimed to assess the variation in different APG parameters in patients with chronic venous disease and to evaluate the reliability of APG in test-retest situations. METHOD: Seventeen patients (18 limbs) with chronic venous disease were recruited into this study. Subjects were asked to undergo tests on two occasions, 1 to 6 weeks apart. Three tests were performed at each visit, and three patients had 10 tests performed at one visit. The coefficients of variation were calculated for repeated measurements and test-retest reliability, and the differences between the means of three tests and the 10 tests were also analyzed. RESULTS: The coefficients of variation for the repeated measurements ranged from 7.5% to 27% for the majority of parameters of APG. The differences between the means of three tests and the means of 10 tests were less than 10% in this study. The coefficients of variation of method error were approximately 10% in test retest measures. CONCLUSIONS: This study has shown that evaluations of calf pump function and venous reflux using APG display variations in repeated measurements and in the test-retest measures. The variations found within patients and on retesting patients on different days suggest that APG is very unlikely to be able to detect small changes in the parameters of venous reflux and calf pump function. It is essential to understand the inherent variation of APG measurements when they are used to assess treatments that are designed to improve venous function. PMID- 9357466 TI - Iatrogenic vascular lesions in extremely low birth weight and low birth weight neonates. AB - PURPOSE: Aggressive treatment has improved the long-term outcome of extremely low birth weight (ELBW) and low birth weight (LBW) neonates, but it has also increased the risk of iatrogenic lesions. The aim of this paper is to evaluate the incidence of vascular injuries observed in the neonatal intensive care unit of our hospital. METHODS: From 1987 to 1994, 2898 neonates were admitted to the neonatal intensive care unit; 335 of them were either LBW or ELBW (11.5%). A review of the charts of these neonates disclosed nine neonates (four male, five female) with vascular lesions (2.6%); the mean gestational age of these patients was 28.7 weeks (range, 24 to 33 weeks), the mean weight at birth was 880 g (range, 590 to 1450 g), and the mean weight at diagnosis was 1825 g (range, 1230 to 2700 g). In the same period, 10 neonates with vascular injuries were reported in the 2563 neonates who weighed more than 1500 g (0.3%). The injuries observed in LBW and ELBW group were arteriovenous fistulas (two bilateral) at the femoral level (six neonates), carotid lesion (one neonate), and limb ischemia (two neonates). Injury was associated with venipuncture in seven neonates, and with umbilical catheter in one; the case of carotid lesion was related to surgical error. No general symptoms were observed. RESULTS: The carotid lesion and five arteriovenous fistulas were repaired by microsurgical techniques; one case of limb ischemia was resolved with thrombolytic drugs, whereas an amputation at the knee level was required in the other after 10 days of medical treatment. One neonate with an arteriovenous fistula was just observed according to the parents' wishes. At clinical and echo-color Doppler follow-up, seven of nine neonates had normal vascular function without sequelae. CONCLUSIONS: In our experience, LBW and ELBW neonates are at greater risk than older neonates of the development of iatrogenic vascular lesions. We advocate aggressive microsurgery, medical treatment, or both to obtain good results and prevent late sequelae. PMID- 9357467 TI - Role of nitric oxide and tumor necrosis factor on lung injury caused by ischemia/reperfusion of the lower extremities. AB - PURPOSE: Acute aortic occlusion with subsequent ischemia/reperfusion (I/R) of the lower extremities is known to predispose to lung injury. The pathophysiologic mechanisms of this injury are not clear. In the present study, we studied the role of tumor necrosis factor (TNF) and nitric oxide (NO) in lung injury caused by lower extremity I/R. METHODS: A rat model in which the infrarenal aorta was cross-clamped for 3 hours followed by 1 hour of reperfusion was used. The rats were randomized into five groups: group 1, aorta exposed but not clamped; group 2, aorta clamped for 3 hours, followed by 1 hour of reperfusion; group 3, 1 mg/kg dexamethasone administered before the aorta was clamped; group 4, 25 mg aminoguanidine, a specific inducible NO synthase (iNOS) inhibitor, administered before the aorta was clamped; and group 5, 2 mg/kg TNFbp, a PEG-ylated dimeric form of the high-affinity p55 TNF receptor I (RI), administered before the aorta was clamped. NO concentration in the exhaled gas (ENO) was measured, as an index of NO production by the lung, in 30 minute intervals during I/R. Serial arterial blood samples for TNF assay were obtained during the course of the experiment. At the end of the experiment, the lungs were removed and histologically examined for evidence of injury. RESULTS: ENO in group 2 increased from 0.7 +/- 0.3 ppb at baseline to 54.3 +/- 7.5 ppb at the end of ischemia and remained stable during reperfusion (54.6 +/- 8.5 ppb at the end of reperfusion). ENO production was blocked by aminoguanidine, by dexamethasone, and by TNFbp given before aortic occlusion. Serum TNF in groups 2, 3 and 4 increased rapidly during early ischemia, reaching its peak value 60 minutes after occlusion of the aorta, then gradually declined to baseline levels at the end of ischemia, and remained low during reperfusion. TNFbp decreased serum TNF concentration significantly when it was given before aortic occlusion. Histologic examination of the lungs at the end of the experiment revealed that aminoguanidine, dexamethasone, and TNFbp had a protective effect on the lungs. CONCLUSIONS: Serum TNF increases rapidly during lower extremity ischemia and causes increased production of NO from the lung by upregulating iNOS. Increased NO is associated with more severe lung injury, and iNOS blockade has beneficial effects on the lung. TNF blockade before ischemia decreases NO production by the lung and attenuates lung injury. ENO can be used as an early marker of lung injury caused by lower extremity I/R. PMID- 9357468 TI - Enhancement of nitric oxide production after arterial reconstruction in patients with arteriosclerosis obliterans. AB - PURPOSE: Nitric oxide (NO) not only relaxes vascular smooth muscles, but it also reduces platelet adhesion and is itself a potent antiaggregatory substance. Experimental studies have shown that the release of NO is modulated by the blood flow. However, little clinical information is available about the effects of hemodynamic changes after arterial reconstruction on NO production. We therefore examined whether the plasma levels of nitrite (NO2-) and nitrate (NO3-) ions increased after arterial reconstruction in patients with arteriosclerosis obliterans (ASO). METHODS: Blood samples were obtained from the femoral artery in seven patients who underwent arterial reconstruction and seven healthy individuals (control). NO2- and NO3- levels were measured using high-performance liquid chromatography before the operation and 1 hour and 14 days after the operation. In addition, the mean femoral artery blood flow and ankle-brachial pressure index (ABI) were also measured using a duplex and Doppler velocimeter both before and after the operations. RESULTS: In the control subjects, the mean plasma NO2-, NO3-, and NOx (NO2- plus NO3-) levels in the femoral artery were 0.37 +/- 0.15 mumol/L, 45.6 +/- 10.8 mumol/L, and 46.0 +/- 10.9 mumol/L, respectively. Before the operation in the patients with ASO, the mean plasma NO3- (23.8 +/- 2.2 mumol/L) and NOx levels (24.0 +/- 2.3 mumol/L) were significantly lower than those in the control subjects, whereas the plasma NO2- levels (0.27 +/ 0.04 mumol/L) were comparable between the two groups. At 14 days after operation, the mean plasma NO3- and NOx levels in the femoral artery were significantly increased to 42.8 +/- 5.6 mumol/L and 43.4 +/- 5.6 mumol/L compared with those before the operation, whereas the mean plasma NO2- levels (0.50 +/- 0.05 mumol/L) changed significantly. The mean ABI and the mean flow rate before the operation were 0.32 +/- 0.07 and 344 +/- 145 ml/min, respectively. Both the ABI and the mean flow rate significantly increased to 1.04 +/- 0.06 and 627 +/- 141 ml/min after the operation. CONCLUSIONS: In patients who have ASO, the mean plasma level of NO is significantly lower than that of healthy individuals. In patients with ASO, the mean blood flow increased significantly after arterial reconstruction. This hemodynamic improvement may thus enhance NO production and may also help to maintain the patency of the bypass graft or native artery. PMID- 9357469 TI - Arterial thrombosis induces early upregulation of intercellular adhesion molecule in the media. AB - PURPOSE: Thrombosed peripheral vessels that are pharmacologically or mechanically recanalized have diminished long-term patency rates compared with vessels that are repaired before occlusion. We hypothesized that thrombosis induces proinflammatory changes in the arterial media that may contribute to postthrombotic vascular remodeling. METHODS: We studied expression of intercellular adhesion molecule (ICAM), a mediator of leukocyte recruitment, in the arterial wall after thrombosis. Thrombosis was induced in rabbit superficial femoral arteries by embolizing polystyrene beads (Thr-emb) or by ligation (Thr lig). Control vessels were dissected but not ligated (C-dis) or were subjected to bead embolization and immediate removal (C-emb). Arterial wall ICAM expression was measured by indirect immunohistochemical analysis at 6 hours, 24 hours, and 1 week. Staining intensity was graded from 0 (none) to 4 (intense) by observers who were blinded to the experimental conditions. RESULTS: No increase in ICAM expression by thrombosed vessels was present at 6 hours. After 24 hours, ICAM expression in the media of thrombosed vessels was increased (Thr-emb, 2.3 +/- 0.5; Thr-lig, 2.0 +/- 0) compared with control vessels (C-dis, 0 +/- 0; C-emb, 0.8 +/- 0.5; p < 0.004). This difference became more marked at 1 week. ICAM staining localized to actin-staining regions of the media. CONCLUSIONS: Arterial thrombosis, but not surgical injury, induces pronounced early and sustained upregulation of ICAM expression in smooth muscle-containing regions of the arterial media. Upregulation of ICAM is likely to promote recruitment of inflammatory cells or mediate vascular remodeling after luminal thrombosis. PMID- 9357472 TI - A new technique for laparoscopic aortobifemoral grafting in occlusive aortoiliac disease. AB - PURPOSE: This article describes an original laparoscopic technique that allows performance of aortobifemoral bypass grafting. METHODS: The technique described is the result of 6 years of in vitro and animal experimentation. It also represents the end result of prior clinical research with laparoscopy-assisted aortoiliac surgery and totally laparoscopic retroperitoneal aortobifemoral bypass grafting. The technique consists of the creation of a flap of retroperitoneum that is used to separate the intraperitoneal organs from the content of the retroperitoneal cavity. Surgery can then be conducted with no intrusion of any intraabdominal organ into the operative field. Another advantage is that the pneumoperitoneum is equally distributed among the two cavities. A conventional aortobifemoral bypass procedure is then performed with laparoscopic instrumentation. RESULTS: The described technique has been performed in three patients to date. The patients' intraoperative blood loss did not exceed 500 ml, and no complication arose. The intraoperative need for crystalloids was of the order of 3 L (almost half the quantity usually administered). The patients' analgesia requirement was low in these patients, and return to walking was rapid. They were sent home between the fourth and sixth postoperative days. CONCLUSIONS: The innovative technique described here is safe and appears to ease the patient's postoperative course. Data recovered from the multicenter study, which is now in its preliminary phase, should help answer numerous questions. We expect the procedure to be reproducible in other university centers that are participating in the trial. PMID- 9357471 TI - Effect of retroviral transduction on human endothelial cell phenotype and adhesion to Dacron vascular grafts. AB - PURPOSE: Retroviral transduction for genetic enhancement of endothelial cell (EC) anti-thrombotic phenotype offers potential for improving the clinical success of vascular graft seeding; however, application of this technique may bring concomitant alteration in cell functionality. METHODS: Human microvascular ECs were transduced with a retroviral vector encoding for the marker gene beta galactosidase. Transduced endothelial cells (rtECs) and nontransduced endothelial cells (ntECs) were evaluated by flow cytometry for expression of intercellular adhesion molecule (ICAM)-1 and tissue factor (TF) on both smooth (coverslips) and graft (Dacron, 6 mm inside diameter) surfaces under static and shear exposed conditions. Graft EC retention was measured after 6-hour pulsatile perfusions. Platelet and neutrophil adherence was measured on perfused coverslips. RESULTS: Lower levels of ICAM-1 were expressed by rtECs on coverslips under both static (p < 0.01 vs static ntECs) and shear exposed conditions (p < 0.01 vs static and shear ntECs). Accordingly, fewer polymorphonuclear leukocytes adhered to rtEC monolayers (p < 0.01 vs ntECs). No difference in ICAM-1 and TF expression by static graft seeded rtECs and ntECs was observed. However, graft-seeded rtECs that were exposed to wall shear stress displayed less TF than sheared ntECs (p < 0.05). Transduction did not affect EC retention to the sheared graft surface. CONCLUSIONS: These data suggest that retroviral transduction does not elicit a prothrombotic/proinflammatory phenotype, rather indices of these states appear in some conditions to be reduced. Further, transduction does not adversely affect EC adherence to Dacron graft surfaces under arterial hemodynamics. PMID- 9357473 TI - Thoracic aorta transobturator bipopliteal bypass as eventual durable reconstruction after removal of an infected aortofemoral graft. AB - A 36-year-old man was referred with aortofemoral graft infection and perigraft duodenal erosion. The aortofemoral graft was removed, and bilateral axillo superficial femoral grafts were constructed. Recurrent failures of these grafts prompted us to convert to a more-durable reconstruction. A straight graft was anastomosed to the lower thoracic aorta, routed retroperitoneally, and attached to an inverted U-shaped bilateral transobturator bypass graft, which was anastomosed to both above-knee popliteal arteries. After 3 years, the patient has remained well and the grafts are patent. This operation represents a durable in line reconstruction that avoids all previously infected areas after removal of an infected aortofemoral graft. PMID- 9357470 TI - Tenascin expression is associated with a chronic inflammatory process in abdominal aortic aneurysms. AB - PURPOSE: This study was performed to test whether tenascin, a large oligomeric glycoprotein of the extracellular matrix, is present in AAA disease and whether it could play a pathophysiologic role in the development of this disease. METHODS: Tenascin immunoreactivity was investigated from samples of the aneurysmal walls of 17 patients with AAAs, and the results were compared with the results of those of six patients with aortoiliac occlusive disease (AOD) and one normal control patient. To study the source of tenascin mRNA synthesis, some tissue samples were also examined with a tenascin RNA probe by in situ hybridization. RESULTS: The difference in immunoreactivity between the AODs and AAAs was especially prominent in the adventitial layer, where the specimens from AAAs displayed strong diffuse and reticular immunostaining. In AAAs the immunostaining was clearly associated with the degree of mononuclear inflammatory cell infiltrate and with the neovascularization of the adventitial layer. CONCLUSION: Tenascin expression is evident in AAA disease and is distinctly associated with mononuclear inflammatory cells. The adhesive properties of tenascin may offer a relevant explanation for the mechanism by which monocytes transmigrate into the aortic wall. The definitive role of tenascin in AAA process may be more complex, however, and will necessitate further investigation. PMID- 9357474 TI - Exclusion of a sciatic artery aneurysm and an obturator bypass. AB - This case report describes surgical treatment in a sciatic artery aneurysm with hypoplastic external iliac and femoral arteries. An obturator bypass grafting procedure from the internal iliac artery to the distal sciatic artery was performed after aneurysmal exclusion was achieved by proximal and distal ligation. This method offers an acceptable option for surgery in some types of sciatic artery aneurysms. PMID- 9357475 TI - Rupture of a nonaneurysmal atherosclerotic infrarenal aorta. AB - We report a case of spontaneous rupture of a nonaneurysmal infrarenal aorta through an area of atherosclerotic thinning. Close follow-up, aggressive antihypertensive therapy, and serial imaging of asymptomatic patients with documented aortic ulceration is warranted. In symptomatic patients, surgical intervention should be undertaken so that transmural aortic rupture can be averted. PMID- 9357477 TI - Salvage of femoropedal bypass graft complicated by interval gangrene and vein graft blowout using a flow-through radial forearm fasciocutaneous free flap. AB - We report the case of a 71-year-old man who had interval gangrene of his calf with subsequent vein graft blowout 3 months after undergoing a femoral-to dorsalis pedis saphenous vein bypass grafting procedure. To provide wound coverage, restore vascular continuity, and preserve functional ambulation, a flow through radial forearm fasciocutaneous free flap was interposed between cut ends of the bypass graft. Venous drainage of the flap was from the cephalic vein to the popliteal vein. At 1 month after the operation, the patient had complete wound healing and began to ambulate. At 11 months an asymptomatic high-grade stenosis in the distal radial artery segment of the reconstruction was successfully treated with percutaneous angioplasty. After 22 months of follow-up there have been no further complications, and the patient continues to have full, functional ambulation. The radial forearm flow-through free flap allows single stage restoration of bypass graft continuity and coverage of extensive, complex tissue defects. This technique represents a novel approach to this difficult problem and provides a viable alternative to major limb amputation. PMID- 9357476 TI - Mucopolysaccharidosis presenting as pediatric multiple aortic aneurysm: first reported case. AB - Within the pediatric population, the rare aortic aneurysm is most often brought on by congenital cardiovascular malformation or connective tissue disorder, trauma, inflammatory disease, or infection. Thus our 8-year-old patient who had multiple aortic aneurysms and evidence of mucopolysaccharidosis presented a doubly unique case. Three and one-half months after the patient underwent emergency aortic valve replacement, we performed resection and graft replacement of both her descending thoracic aorta and thoracoabdominal aorta. Histologic analysis of the aneurysm wall displayed severe medial degeneration with large deposits of acid mucopolysaccharides. Subsequent evaluation, although negative for connective tissue disorders, showed glycosaminoglycans, chondroitin sulfate, and heparan sulfate in the patient's urine. These findings are diagnostic for a heterogeneous group of storage diseases termed mucopolysaccharidoses, although testing of the patient's cultured fibroblasts failed to reveal any specific previously described enzymatic defect. After reviewing the literature, we believe that this is the first known successfully treated pediatric aortic aneurysm associated with mucopolysaccharidosis. PMID- 9357480 TI - Demographic transition poses a challenge to societies worldwide. PMID- 9357478 TI - Noninvasive vascular imaging in the diagnosis and treatment of adventitial cystic disease of the popliteal artery. AB - This brief case report describes the successful outcome after surgical excision of multiple adventitial cysts of the popliteal artery in a 75-year-old man with rapidly worsening claudication. It highlights several unsettled points concerning the diagnosis, cause, and management of cystic adventitial disease of the popliteal artery and compares duplex ultrasound, computed tomography, and magnetic resonance angiography in the noninvasive diagnosis and treatment of this condition. PMID- 9357479 TI - Regarding "Iliac vein compression syndrome: case report and review of the literature". PMID- 9357481 TI - The role of the Federation of European Societies for Tropical Medicine and International Health. PMID- 9357482 TI - A bioassay for evaluating antimalarial activity and for measuring concentration in plasma. AB - Our objective was to develop a bioassay method based on the isotopic microtest of Desjardins and to validate it by comparing the results with high performance liquid chromatograph (HPLC). The bioassay was developed using a continuous culture of P. falciparum, and the test material consisted of 32 human serum samples from 22 Filipino patients treated only with chloroquine (CQ) for noncomplicated malaria. A blind assay of the serum was done using the Desjardins method and HPLC. A good correlation (r2 = 0.90, n = 32) was observed between the CQ plus monodesethylchloroquine (DCQ) concentrations obtained by HPLC and the equivalent CQ by bioassay. It was also shown that the antimalarial activity of CQ is practically identical to that of its main metabolite DCQ, the activity of DCQ being 0.9 times the activity of CQ. The bioassay is reliable, semi-automated, and reproducible. This method may be used as a complementary technique to HPLC for determining biological activity in serum and for measuring the kinetics of this activity, as well as to indicate the existence of new metabolites. The bioassay can be applied to other protozoans for which an in vitro model is available. PMID- 9357484 TI - Compliance with and tolerance of mefloquine and chloroquine + proguanil malaria chemoprophylaxis in French short-term travellers to sub-Saharan Africa. AB - To compare the compliance with and tolerance of mefloquine (MQ) and chloroquine + proguanil (CQ-PRO) chemoprophylaxis, we conducted a study using a self-reported questionnaire in 2 groups of native French adult visitors to Senegal or Kenya. CQ (100 mg daily) + PRO (200 mg daily) prophylaxis was prescribed for all patients travelling to Senegal and for those going to Kenya when MQ was contraindicated; MQ (250 mg weekly) was prescribed for the other subjects. There were no significant differences in age, sex, exposition and measures of protection against mosquito bites, concomitant drug use or mean duration of chemoprophylaxis between the 2 groups, and compliance during travel was excellent in both. Chemoprophylaxis was more frequently interrupted prematurely in the MQ group. The rates of overall side-effects attributed to malaria chemoprophylaxis were 16% for MQ against 12% CQ-PRO (not significant). However, nonserious neuropsychiatric adverse events are more frequent with MQ: 11.5% compared to 2% with CQ-PRO. MQ should be used with caution. PMID- 9357483 TI - Emerging viral pathogens in long-term expatriates (II): Dengue virus. AB - Dengue virus infections have been well known for many years; still dengue virus is regarded as an 'emerging' pathogen, as the disease profile is changing. Its geographical range and overall incidence, and the incidence of the associated complications, dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS), are on the increase. Modern-day travel and increasing urbanization seem to be the main contributing factors. In order to estimate the risk of infection during long term stays in dengue-endemic countries, we tested sera obtained from 323 development aid workers and their family members who had spent on average 9.8 years in dengue-endemic regions for the presence of dengue virus antibodies. Dengue virus antibody screening was done by a commercially available immunofluorescence test (IF). Reactive samples were re-tested by an in-house IF and also tested for cross-reactivity to yellow fever virus using yellow fever IF and neutralization test (NT). Evaluation of the results revealed that the screening test has a specificity of at least 63.2%. In 12 of 19 initially positive cases crossreacting antibodies against yellow fever virus could be ruled out. Three cases remained indeterminable, whereas four of the reactive and 10 (out of 12) of the borderline reactive cases showed crossreactivity with yellow fever virus, probably due to previous vaccination. We found seroprevalence rates of 4.3% with no significant differences related to gender or area of upbringing. Seroprevalence rates were evaluated according to region of suspected or confirmed infection. In two cases the dengue infection had taken a classical clinical course; in another three cases an extraordinary febrile illness was reported in the history. None of the other seropositive individuals had a history of an illness possibly attributable to dengue virus infection. Our results show that there definitely is a risk for long-term expatriates to acquire (mostly non- or oligo-symptomatic) dengue infection, which might be important especially in the light of the supposed aetiology of DHF or DSS as a secondary infection with another dengue virus serotype. PMID- 9357485 TI - The pharmacokinetics of a single dose of artemisinin in subjects with liver cirrhosis. Bach Mai-Amsterdam Research Group on Artemisinin. AB - Artemisinin is mainly eliminated by hepatic transformation. To investigate whether the clearance of artemisinin in patients with liver cirrhosis is different from healthy volunteers, a pharmacokinetic study was performed in male Vietnamese patients with Child B cirrhosis of the liver who received 500 mg of artemisinin orally. The results were compared to those found in a previous study in healthy subjects. The mean (+/- SD) area under the concentration time curve was 2365 (+/- 1761) h ng/ml; the mean (+/- SD) clearance, 382 (+/- 303)L/h. The elimination half life was 4 (+/- 1.3) h extimated by log-linear regression and 2.4 +/- 0.9 h estimated by non-linear regression using a one-compartment first order elimination model. The mean (+/- SD) absorption time was 1.55 (+/- 0.8) h. These results were not different from the results of healthy subjects and show that liver disease has no effect on the availability and clearance of oral artemisinin, indicating that artemisinin has an intermediate hepatic extraction ratio and that there is no significant first pass effect. PMID- 9357486 TI - Leopard or chameleon? The changing character of international health economics. AB - Over the last 25 years the discipline of health economics has developed substantially. As an applied discipline, it has adapted and changed over time in response to the changing concerns of policy-makers, planners and managers. This paper questions whether it is like a chameleon, changing its appearance in response to the external environment, or like the leopard that never changes its spots. In answering the question, the paper presents an overview of the development of health economics as it has been applied in low and middle income countries distinguishing three eras, the 1970s, 1980s, and 1990s, and argues that in each of these eras the preoccupations of health economists have been somewhat different. In each era the key contributions of health economics are identified. The paper ends by considering future research priorities, and the obligations of developed country institutions in terms of research topics and mode of work. PMID- 9357488 TI - Successful control of onchocerciasis with community-based ivermectin distribution in the Rio Santiago focus in Ecuador. AB - Onchocerciasis is a major blinding disease in equatorial Africa and Central and South America. Ivermectin is a safe and effective drug in the treatment of this disease and now forms the basis of disease control in most endemic areas. We report the findings of long-term control of this infection in the Rio Santiago focus in Ecuador, between January 1990 and December 1996, using a strategy of giving ivermectin treatments biannually in hyperendemic communities and annually in meso- and hypoendemic communities. Ivermectin was administered by local health workers from each community. A high level of compliance to ivermectin was achieved, with 81.9% to 98.0% of those eligible receiving the drug at each treatment instance. The impact of ivermectin therapy was monitored using a cohort of 120 randomly selected infected individuals from 8 hyperendemic communities. The geometric mean microfilarial density of this group declined from 19.3 to 0 mf/mg over the 84-month observation period. Ivermectin had a significant impact on anterior segment ocular disease, acute onchodermatitis and sowda. The rate of infection of blackflies declined from 1.1% in 1989-0.08% in 1996, which is below the vectorial capacity of the Simulium vector and, as no new nodules were detected after 1994 and no children under 5 became infected over the observation period, it is likely that the transmission of this infection was interrupted in the study area. PMID- 9357487 TI - Evaluation of a commercial immunodiagnostic kit incorporating lipoarabinomannan in the serodiagnosis of pulmonary tuberculosis in Ghana. AB - We evaluated 'Mycodot', a commercially marketed immunodiagnostic test for tuberculosis which detects antibodies to lipoarabinomannan antigen. Serum was tested from 52 patients with newly diagnosed smear-positive pulmonary tuberculosis, of whom 20 were HIV-positive and 32 HIV-negative. Control sera were taken from 40 patients of whom 20 had acute non-tuberculous lobar pneumonia and 20 patients had no respiratory disease. The test was found to have a very high specificity of 97.5% (95% CI:92.5-100%). However, the sensitivity in HIV-negative patients was 56% (95% CI:39-73%), and was substantially lower at 25% (95% CI:6 44%) in HIV-positive patients. IN CONCLUSION: 'Mycodot' was found to be a highly specific and easily performed assay. However, the poor sensitivity, especially in HIV-infected patients, renders it unlikely to be useful either as a primary or adjunctive diagnostic test for tuberculosis, particularly in countries with a high prevalence of HIV. A larger trial of this assay in Ghana was not deemed necessary. PMID- 9357489 TI - Studies of the hepatic mitochondrial and microsomal mixed-function oxidase system during Plasmodium yoelii infection and inducer treatment in Swiss albino mice. AB - Plasmodium yoelii infection resulted in depression of hepatic mitochondrial and microsomal mixed-function oxidase system indices, e.g. cytochrome P-450, cytochrome b5 and phase II detoxification enzyme glutathione-S-transferase, while heam and haemozoin registered a marked increase in Swiss albino mice. Phenobarbitone (inducer) treatment showed induced levels of hepatic mitochondrial and microsomal cytochrome P-450 and glutathione-S-transferase in normal as well as in infected mice. The induced cytochrome P-450 and glutathione-S-transferase activities were similar in normal and infected mice. The findings were further supported by the isoenzymic profile and drug-binding properties of the terminal monoxygenase, cytochrome P-450. PMID- 9357490 TI - The need for research on dementia in developing countries. AB - Dementia is one of the commonest and most disabling late-life mental disorders. Its prevalence and incidence have been assessed in developed countries, and show little geographical variation between countries and regions. Although most older people live in developing countries, little research has been carried out in those settings. There is some evidence that age-specific prevalence rates for dementia in developing countries may be relatively low. More research is needed to allow developing countries to estimate the current extent, type and cost of medical and social service provision, and to make confident predictions of future need. Research in different cultures with different levels of economic and industrial development will also increase the variance of environmental exposure, facilitating the identification of environmental risk factors and gene environment interactions for dementia. Research methodologies need to be adapted to the different circumstances of developing countries, with implications for sampling, cognitive screening and definitive dementia diagnosis. PMID- 9357491 TI - Poverty and mortality among the elderly: measurement of performance in 33 countries 1960-92. AB - This paper analyses the effect of income and education on life expectancy and mortality rates among the elderly in 33 countries for the period 1960-92 and assesses how that relationship has changed over time as a result of technical progress. Our outcome variables are life expectancy at age 60 and the probability of dying between age 60 and age 80 for both males and females. The data are from vital-registration based life tables published by national statistical offices for several years during this period. We estimate regressions with determinants that include GDP per capita (adjusted for purchasing power), education and time (as a proxy for technical progress). As the available measure of education failed to account for variation in life expectancy or mortality at age 60, our reported analyses focus on a simplified model with only income and time as predictors. The results indicate that, controlling for income, mortality rates among the elderly have declined considerably over the past three decades. We also find that poverty (as measured by low average income levels) explains some of the variation in both life expectancy at age 60 and mortality rates among the elderly across the countries in the sample. The explained amount of variation is more substantial for females than for males. While poverty does adversely affect mortality rates among the elderly (and the strength of this effect is estimated to be increasing over time), technical progress appears far more important in the period following 1960. Predicted female life expectancy (at age 60) in 1960 at the mean income level in 1960 was, for example 18.8 years; income growth to 1992 increased this by an estimated 0.7 years, whereas technical progress increased it by 2.0 years. We then use the estimated regression results to compare country performance on life expectancy of the elderly, controlling for levels of poverty (or income), and to assess how performance has varied over time. High performing countries, on female life expectancy at age 60, for the period around 1990, included Chile (1.0 years longer life expectancy), China (1.7 years longer), France (2.0 years longer), Japan (1.9 years longer), and Switzerland (1.3 years longer). Poorly performing countries included Denmark (1.1 years shorter life expectancy than predicted from income), Hungary (1.4 years shorter), Iceland (1.2 years shorter), Malaysia (1.6 years shorter), and Trinidad and Tobago (3.9 years shorter). Chile and Switzerland registered major improvements in relative performance over this period; Norway, Taiwan and the USA, in contrast showed major declines in performance between 1980 and the early 1990s. PMID- 9357493 TI - Growth hormone, skin and wound healing--experimental studies in the rat. PMID- 9357492 TI - Cellular and molecular effects of growth hormone and estrogen on human bone cells. AB - The aim of the present thesis is to examine some aspects of the biological effects of growth hormone (GH) and estrogen on bone cells in vitro. The first part of the thesis describes characterization of model systems to study normal human osteoblasts and osteoclasts in vitro. Three culture systems for human osteoblasts have been characterized, representing different stages of osteoblasts differentiation/maturation: (1) mature osteoblasts cultured from trabecular bone explants (trabecular osteoblasts), (2) less mature osteoblasts (stromal osteoblasts) cultured from bone marrow and (3) osteoblast precursor cells cultured also from bone marrow. This classification is based on quantitative and qualitative differences in the expression of osteoblast phenotypic markers by these cells. These systems are useful in studying the regulation of hormones and growth factors of different stages of osteoblast differentiation. Further characterization of the osteoblast differentiation pathway is still needed, especially the identification of surface markers that can definitively identify intermediate stages of osteoblast differentiation. Normal human osteoclasts were cultured from bone marrow mononuclear cells and exhibited the main characteristics of osteoclast phenotype: production of tartrate-resistant acid phosphatase, presence of a ruffled border and the ability to resorb mineralized matrix. This model is useful for studying factors regulating osteoclast commitment and differentiation. The second part of the thesis deals with the effects of GH on proliferation and differentiation of trabecular and stromal osteoblasts. Effects of GH on human osteoblasts were dependent on their degree of maturation. GH stimulated cell proliferation in both trabecular and stromal osteoblasts. While it increased the functional activity of trabecular osteoblasts, these effects were absent in stromal osteoblast cultures. Human trabecular osteoblasts produce mainly IGF-II, IGFBP-3 and minute quantities of IGF-I in culture. GH does not seem to regulate the local production of IGF-II or IGFBP-3. However, IGFs and their binding proteins may exert important regulatory effects on the biological effects of GH on human osteoblasts, and this role needs to be studied. Sincer GH exerted profound effects on the biological functions of human osteoblasts in vitro, the hypothesis that either decreased production of GH or decreased sensitivity of bone cells to its action leads to bone loss and osteoporosis was examined. No differences in the basal or GH-stimulated production of IGF-I, IGF-II or IGFBP-3 were found between osteoporotic patients and age-matched normals. Similarly, The response of bone cells to in vivo and in vitro stimulation by GH was similar in the two groups. These studies do not support the hypothesis of the presence of major defects in production of GH or the presence of resistance to its effects in bone cells in patients with osteoporosis. The last part of the thesis deals with the cellular mechanisms mediating estrogen actions on bone cells. Thus, the potential role of estrogen regulation of bone-resorbing cytokines in the pathogenesis of bone loss in postmenopausal women was examined. Estrogen was found to inhibit IL-6 production and gene expression in an immortalized human osteoblastic cell line expressing high levels of estrogen receptors (hFOB/ER9 cell line). To further examine the relationship between estrogen deficiency and early postmenopausal bone loss, levels of IL-6, IL-6sR, IL-1 alpha, IL-1 beta and IL-1ra were measured in bone marrow plasma and bone marrow cultures obtained from 40 postmenopausal women, half of whom were on estrogen replacement therapy. No difference was found between the groups in any of these parameters. This suggests that neither IL-1 nor IL-6 by itself is the major mediator for increased bone loss due to estrogen deficiency in the early postmenopausal period. (ABSTRACT TRUNCATED) PMID- 9357494 TI - Complications of injectable synthetic polymers in facial augmentation. AB - BACKGROUND: Injectable synthetic materials have been used for augmentation of soft tissue defects, correction of wrinkles, and augmentation of facial features such as the nasal dorsum. Success has been limited by inflammatory reactions, material migration, and the difficulty of removal should complications occur. OBJECTIVE: To evaluate complications resulting from soft tissue augmentation with injectable alloplastic materials. METHODS: Retrospective review of seven cases. Clinical history, treatment, histopathologic findings, and outcomes are assessed. RESULTS: Inflammatory reaction and tissue damage were refractory to antibiotics and steroids, and surgery was required to remove the foreign material. Histologic examination revealed giant cell foreign body reaction in all cases. CONCLUSION: Injectable synthetic polymers can produce significant complications including deformity and inflammatory tissue destruction, the control of which is complicated by the difficulty of removing the materials. Removable tissue fillers, such as e-PTFE, or natural materials such as collagen, autologous, fat, or Alloderm, should be considered instead. PMID- 9357495 TI - Resurfacing of pitted facial scars with a pulsed Er:YAG laser. AB - BACKGROUND: Laser resurfacing has beneficial effects for the treatment of several skin conditions. Recently, the pulsed Er:YAG laser has been shown to be a highly effective treatment for several kinds of pitted facial scars. OBJECTIVE: The purpose of this study was to assess the efficacy and safety of pulsed Er:YAG laser skin resurfacing for pitted facial scars. METHODS: Four patients with small pox scars, five patients with chicken pox scars, and 21 patients with acne scars were included in this study. All patients were skin type III and IV. All patients were instructed to use tretinoin cream 0.05% nightly for 2-4 weeks prior to the laser treatment. The pulsed Er:YAG laser with 2-mm handpiece at the setting of 500 mJ/pulse, 3.5-4.5 W was used. Two weeks after laser treatment, topical application of hydroquinone 4%, tretinoin 0.05%, and hydrocortisone 1% cream was recommended for 2-4 weeks. Facial photographs were obtained at baseline and 2 week intervals postoperatively with a 35-mm single lense reflex camera equipped with a lense mounted ring flash. The results of treatment were evaluated for the changes of skin texture and color at 2 weeks, 1 month, and 3 months. Three patients with acne scars agreed to skin biopsy. RESULTS: Three months after laser treatment, all patients with small pox and chicken pox scars were improved about 55%, and patients with acne scars were improved about 40% on average. CONCLUSION: Pulsed Er:YAG laser skin resurfacing is an effective and safe treatment for pitted facial scars. PMID- 9357496 TI - The carbon dioxide laser. An alternative for the treatment of actinically damaged skin. AB - BACKGROUND: The treatment of complex and diffuse actinic keratoses involving the face presents a problem in that they frequently recur despite traditional treatment modalities. The carbon dioxide (CO2) laser is an effective method for resurfacing actinically damaged facial skin. OBJECTIVE: The purpose of this study is to show the usefulness of the CO2 laser for the treatment of actinically damaged skin in patients with proven actinic keratoses and squamous cell carcinoma in situ of the face. METHODS: In an office surgery setting, the Sharplan 1030 or 40C CO2 laser with the SilkTouch flashscanner attachment was utilized to treat various regions of the face in 14 patients. RESULTS: All patients were satisfied with the aesthetic outcome of their laser procedures and no clinical evidence of residual or recurrent lesions have been noted. There were no long-term complications reported. CONCLUSIONS: Based on this preliminary report, the CO2 laser appears to be an excellent alternative for the surgical treatment of premalignant lesions of the face and can be used effectively without significant complications. PMID- 9357497 TI - Increased smooth muscle actin, factor XIIIa, and vimentin-positive cells in the papillary dermis of carbon dioxide laser-debrided porcine skin. AB - BACKGROUND: Pulsed carbon dioxide (CO2) laser debridement is now being used as therapy for photodamaged skin. It has been proposed that the long duration of erythema and a tissue scaffold, which results from tightening of the collagen helix induced by the laser heat, may lead to tightening of sagging skin and skin creases of lesser magnitude. METHODS: Weanling pigs exposed to mild and moderate erythema producing doses of sulfur mustard (bis-2-chloroethyl sulfide; HD) were treated with the CO2 laser (Tru-Pulse) at 6, 24, and 48 hours after exposure. In addition to histologic examination of laser-debrided and nondebrided biopsy specimens obtained at 14 days after exposure, immunohistochemical staining with antibodies to smooth muscle actin, Factor XIIIa, vimentin, and CD3 was performed. RESULTS: CO2 laser debridement of the HD-exposed skin resulted in clearing of the cytologic atypia induced by this chemical carcinogen and reduced the inflammatory infiltrate. In addition laser debridement resulted in increased numbers of stromal cells within the papillary dermis, which showed immunohistochemical staining for smooth muscle actin, Factor XIIIa, and vimentin. CONCLUSIONS: CO2 laser debridement is effective in clearing the epidermis of cytologically damage cells in HD as well as solar-damaged skin. In addition CO2 laser debridement may result in tightening of sagging skin and produce a decrease in skin creases initially, by inducing increased stromal cells within the papillary dermis, with prominent contractile actin filaments. The collagen produced by these stromal cells may subsequently maintain these improvements in the photoaged skin. PMID- 9357498 TI - Superficial congenital compound melanocytic nevus. Another pitfall in the diagnosis of malignant melanoma. AB - METHODS: We reviewed the histology of 42 superficial congenital compound melanocytic nevi and found features that simulate malignant melanoma. In some, there were asymmetry and poor circumscription, an increased number of single melanocytes, which predominated over nests of melanocytes in some high power fields, single melanocytes were not equidistant from one another, scatter of single melanocytes was above the dermoepidermal junction, and there was confluence of nests of melanocytes, which are features in common with a malignant melanoma. RESULTS: Fifty-six percent of the cases had three to four criteria present, and 92% of the cases had three to six criteria present. The mean age of the patients at the time of biopsy was 20 years, and the mean size of the nevus at time of biopsy was 1.7 cm. CONCLUSIONS: Appreciation that simulation of a melanoma in situ by the epidermal component of a superficial congenital compound melanocytic nevus in older children and adults is an expected finding and may be similar to congenital nevi biopsied shortly after birth has not been emphasized. Awareness of these simulants may prevent misinterpretation of a benign superficial congenital compound melanocytic nevus as a malignant melanoma. PMID- 9357499 TI - Laser skin resurfacing with the Q-switched Nd:YAG laser. AB - BACKGROUND: Recent studies have shown that char-free pulsed carbon dioxide lasers are useful in the treatment of rhytids. Other infrared lasers have also been observed to induce changes in the skin. OBJECTIVE: In this pilot study, we evaluated the rhytid resurfacing capability of the Q-switched neodymium:yttrium aluminum-garden (QS Nd:YAG) laser at 1064 nm as compared with char-free carbon dioxide lasers at 10,600 nm. METHODS: Three lasers were used in the study: The QS Nd:YAG laser, the SilkTouch carbon dioxide laser, and the UltraPulse carbon dioxide laser. Eleven subjects were treated in either the periorbital or perioral region with the QS Nd:YAG laser on one side of the face, and both of the char free pulsed carbon dioxide lasers on the other side. The subjects were evaluated at posttreatment days 7, 30, 60, and 90 for improvement of rhytids, healing, pigmentary changes, and erythema. RESULTS: All 11 patients treated with the char free carbon dioxide lasers improved. Nine of 11 patients treated with the QS Nd:YAG laser were improved. Healing (complete reepithelialization) was noted to occur 3-6 days earlier in sites treated with the QS Nd:YAG than in sites treated with char-free carbon dioxide lasers. Pigmentary changes were not observed in any treatment site. Erythema was observed at 1 month after treatment in all areas treated with the char-free carbon dioxide lasers, but only three patients treated with the QS Nd:YAG exhibited erythema. These were the same three QS Nd:YAG treated patients whose clinical improvement was comparable with that of the char free carbon dioxide lasers. CONCLUSION: The Q-switched Nd:YAG laser may play a role in the treatment of rhytids. PMID- 9357500 TI - Long-term epilation using the EpiLight broad band, intense pulsed light hair removal system. AB - BACKGROUND: The long-term epilation of hair is the goal of several lasers and intense pulsed light systems. OBJECTIVE: The purpose of the study is to use the EpiLight Hair Removal System to assess long-term epilation and to assess its safety profile following a single treatment session. METHODS: Patients received a single treatment with the Epilight Hair Removal System after entering the patient's skin type, skin color, hair color, and hair density into the system's computer software. Treatment parameters include various wavelengths of light, pulse duration, pulse delay, and energy fluence. Thirty-seven subjects received a single treatment using one of four cut-off filters consisting of two to five pulses with energies of 34-55 J/cm2. RESULTS: The results of a single treatment show hair clearances occurring immediately and over a 12-week study period. Approximately 60% hair removal was noted at 12 weeks. CONCLUSIONS: The EpiLight Hair Removal System is an effective and safe method for long-term epilation of unwanted hair. PMID- 9357501 TI - Rotation flaps to reconstruct nasal tip defects following Mohs surgery. AB - BACKGROUND: Skin cancer involving the nasal tip is fairly common. The repair of these nasal tip defects is often a challenging opportunity. The current range of reconstructive techniques include partial closure and second intention healing to skin grafts and midline forehead flaps. OBJECTIVE: Here we present an interesting method of nasal tip repair harvesting a rotation flap that uses adjacent skin from lateral and upper bridge of nose. METHODS: Using three case studies examples of the rotation flap are demonstrated to reconstruct the nasal tip defects following Mohs surgery for skin cancer. RESULTS: For medium size defects of up to 1 cm in diameter, rotation flaps offer an alternative surgical one-stage method of reconstruction of the nasal tip in selected cases. CONCLUSION: Satisfactory results can often be achieved with a modest amount of planning, intra-, and postoperative care. Rotation flaps possess a circular configuration that allow recapitulation of the natural creases and contours of nasal tip anatomy resulting in aesthetic repairs. PMID- 9357502 TI - Intralesional electrodesiccation of syringomas. AB - BACKGROUND: Syringomas are common, benign adnexal tumors. In the periorbital area, they pose a cosmetic dilemma for both patients and physicians alike. Many different therapeutic modalities potentially can cause scarring, and recurrences are common. OBJECTIVE: To develop a treatment method that minimizes scarring and subsequent recurrences. METHOD: Each syringoma is treated with short bursts of high frequency low voltage electrodesiccation delivered with a fine needle electrode that is inserted into the center of the syringoma, as deeply as the reticular dermis. RESULTS: Twelve patients treated over a 4-year period showed no permanent adverse effects postoperatively and no recurrences. CONCLUSIONS: Intralesional electrodesiccation is a safe, nonscarring and, reliable method that can be used to eradicate periorbital syringomas. PMID- 9357504 TI - Merkel cell carcinoma. Comparison of Mohs micrographic surgery and wide excision in eighty-six patients. AB - BACKGROUND: Merkel cell carcinoma is an uncommon malignant tumor of the skin that, after standard surgical excision, tends to recur locally and develop regional nodal spread. OBJECTIVE: This study evaluated the use of Mohs micrographic surgery for this aggressive neoplasm. METHODS: A retrospective study of 86 patients with Merkel cell carcinoma established rates of local persistence and the development of regional metastasis after standard surgical excision. Detailed follow-up was available on a subgroup of 13 patients treated with Mohs surgery. RESULTS: Standard surgical excision for local disease was associated with high rates of local persistence (13 of 41 [31.7%]) and regional metastasis (20 of 41 [48.8%]). Mean follow-up was 60 months. Mean follow-up for the group treated with Mohs was 36 months. Only one of 12 (8.3%) Mohs-treated patients with histologically confirmed clearance has had local persistence of disease. This patient underwent a second Mohs excision and has remained disease free for 84 months. Regional metastasis developed in four of 12 cases (33.3%). Regional metastasis developed in none of the four patients treated with radiotherapy after Mohs surgery and in four of eight patients treated with Mohs surgery without postoperative radiotherapy. CONCLUSION: Mohs surgery compares favorably with standard surgical excision. Radiotherapy after Mohs surgery may further reduce persistent metastases in transit and nodal disease. PMID- 9357503 TI - Eccrine porocarcinoma. Report of nine cases. AB - BACKGROUND: Eccrine porocarcinoma is a rare malignancy of the eccrine sweat gland and usually has a long-standing growth on a lower extremity. OBJECTIVE: The aim of this review was to analyze clinical and histopathological findings of eccrine porocarcinoma. METHODS: We report nine cases of eccrine porocarcinoma that we have seen during last 10 years. RESULTS: Eccrine porocarcinoma affects elderly patients in both sexes and is usually found on the lower extremities, but in our review it is similar on the head. The mean size was 1.9 cm in our cases. Elective primary treatment was excision and we did not perform elective lymph node dissection. We have not found evidence of metastases in any of our cases. PMID- 9357505 TI - Pressure differences of elastic compression stockings at the ankle region. AB - BACKGROUND: Elastic compression stockings are widely used medical devices for the prevention of edema and treatment of venous diseases. Many beneficial effects have been described in the past. The exact amount of pressure underneath elastic compression stockings at different areas of the leg remains controversial. OBJECTIVE: To examine the pressure at the skin underneath class II elastic compression stockings at different places around the ankle at the B level. METHODS: Patients with known venous insufficiency and regularly using class II elastic therapeutic compression stockings (25-35 mm Hg) for venous diseases were included. All subjects were wearing completely new stockings at the time of the study. Measurements were performed with an electropneumatic interface pressure measuring device (Oxford Pressure Monitor MK II) with inflatable small sensoring cells. Six sensors were placed around the smallest circumference at the ankle (B area). The highest and lowest recordings as well as the means were evaluated. RESULTS: The mean pressure of class II elastic stockings measured around the ankle was 24.7 mm Hg (SD, 8.4). The lowest pressures were found at the medial site (18.3 mm Hg; 74% of the mean), and the highest at the pretibial zone (33.9 mm Hg, 137%). CONCLUSIONS: The pressure exerted by pressure class II elastic compression stockings a the medial site just above the ankle (B area) is too low to have influence and improve the venous insufficiency. For phlebology this is the target area. It might be a reason for the high recurrence rate of venous ulcers even if patients wear stockings. Also, the mean pressure of class II stockings was found to be below the normal levels of its pressure class (25-35 mm Hg according to the European CEN classification). The results advocate the use of pressure class III elastic compression stocking more often and the use of pelottes or foam pads. PMID- 9357506 TI - Treatment of essential telangiectasias with an intense pulsed light source (PhotoDerm VL) AB - BACKGROUND: The flashlamp-pumped pulsed dye laser (585 and 577 nm) has proven to be a very effective and safe treatment option in the therapy of essential telangiectasias (ETE). Nevertheless, the postoperative intracutaneous hematomata, which most patients see as cosmetically disfiguring, always has been a matter of concern. OBJECTIVE: To test the efficacy and safety of a new, large spot size, intense pulsed light source, the PhotoDerm VL, which omits noncoherent light adjustable within the 515-1200-nm range, in the treatment of ETE. METHODS: Fourteen patients were treated with the PhotoDerm VL. They suffered from ETE of the face, postoperative teleangiectasis of the nose, ETE of both legs, and poikiloderma of Civatte. RESULTS: All treated lesions could be abrogated with excellent results by this new device. There were no unpleasant side effects of the treatment. Additionally, due to the large spot size (2.8 cm2), a larger area could be treated within one session. No anesthesia was required. CONCLUSION: The PhotoDerm VL is an innovative, highly effective, and comparably safe therapeutic alternative to the laser in the treatment of ETE. The rate of cosmetically relevant side effects is considerably smaller, the patient compliance is excellent, and the method can be applied easily in an outpatient setting. PMID- 9357507 TI - Complications of ambulatory phlebectomy. AB - BACKGROUND: Ambulatory phlebectomy, as described by Muller, is a remarkable esthetic, effective, and cost-sparing technique for definitive removal of varicose veins. As this technique is becoming more and more popular, potential complications and incidents have to be recognized by all phlebologists. OBJECTIVE: To review all possible complications after ambulatory phlebectomy and establish their frequency, relevance, treatment, and prevention. METHODS: Extensive review of the European and American literature devoted to phlebectomy, with particular consideration of the complications, are discussed in comparison with the author's personal experience. CONCLUSIONS: Notable adverse incidents after ambulatory phlebectomy are rare. Minor inconveniences are common, depending partially on surgical indications, operator's skill, and experience. Adequate training allows one to minimize untoward reactions. A great risk of ambulatory phlebectomy is the presumed facility of this surgical technique combined with its easy accessibility to poorly trained physicians in phlebology and dermatologic surgery. PMID- 9357508 TI - Pericapillary fibrin cuffs in venous disease. A reappraisal. AB - BACKGROUND: Over the last several years, a number of hypotheses have been proposed to explain the pathogenesis of venous ulceration. According to an early model suggested by Browse and Burnand, pericapillary fibrin cuffs, developing as a result of venous hypertension and extravasation of fibrinogen, act as a barrier to the diffusion of oxygen and nutrients; ultimately, tissue anoxia, cell death, and ulceration would follow. More recent hypotheses include the idea that macromolecules leaking from the vasculature trap growth factors and adhesion molecules, and the notion that white blood cells adhere to and damage endothelial cells in the microcirculation. OBJECTIVE: To review the available evidence for or against a pathogenic role of fibrin cuffs, and hope to provide a useful perspective for this controversial topic. METHODS: We have reviewed the different hypotheses for venous ulceration and the evidence for and against fibrin cuffs acting as a barrier. CONCLUSIONS: Venous ulceration is likely to be the result of a number of distinct pathogenic events. Pericapillary fibrin cuffs remain a prominent feature, whether they act as a barrier, a marker for endothelial cell damage, or as part of an overall mechanism of macromolecular leakage and trapping. PMID- 9357509 TI - Successful treatment of cutaneous Kaposi's sarcoma by the 585-nm pulsed dye laser. AB - The clinical appearance of Kaposi's sarcoma (KS) can cause significant disfigurement and lead to functional impairment, particularly if the lesions ulcerate and become secondarily infected. We describe a patient with a KS plaque on the face that was successfully treated with 585-nm pulsed dye laser (PDL) therapy. No recurrence of the tumor was noted 12 months after the final laser treatment. PMID- 9357510 TI - Granular cell dermatofibroma. A benign tumor that can simulate malignancy. AB - We report on an unusual dermatofibroma with granular cells. The dermatofibroma contained mitotic figures raising a differential diagnosis that included malignant granular cell tumor. Granular cell dermatofibroma is an uncommon variant, but one that clinicians and surgeons should become aware of to ensure accurate diagnosis. PMID- 9357511 TI - Stump the experts. Malignant hidroacanthoma in situ. PMID- 9357512 TI - Dibasic benzo[b]thiophene derivatives as a novel class of active site-directed thrombin inhibitors. 1. Determination of the serine protease selectivity, structure-activity relationships, and binding orientation. PMID- 9357513 TI - 6-Chloro-5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxy]-5-pyridyl]carbamoyl]- indoline (SB-242084): the first selective and brain penetrant 5-HT2C receptor antagonist. PMID- 9357515 TI - Selective, orally active 5-HT1D receptor agonists as potential antimigraine agents. PMID- 9357514 TI - 3-[2-(Pyrrolidin-1-yl)ethyl]indoles and 3-[3-(piperidin-1-yl)propyl]indoles: agonists for the h5-HT1D receptor with high selectivity over the h5-HT1B subtype. PMID- 9357516 TI - Bicyclo[2.2.1]heptane and 6,6-dimethylbicyclo[3.1.1]heptane derivatives: orally active, potent, and selective prostaglandin D2 receptor antagonists. PMID- 9357517 TI - Preparation, characterization, DNA binding, and in vitro cytotoxicity of the enantiomers of the platinum(II) complexes N-methyl-, N-ethyl- and N,N-dimethyl (R)- and -(S)-3-aminohexahydroazepinedichloroplatinum(II). AB - A series of chiral diaminedichloroplatinum(II) complexes derived from [Pt(ahaz)Cl2] (ahaz = 3-aminohexahydroazepine) have been synthesized and tested for cytotoxic activity. Novel synthetic pathways were developed to produce the structural derivatives of the ahaz ligand, with alkyl substituents on the exocyclic nitrogen atom. The platinum(II) complexes of these ligands were synthesized and characterized by NMR and CD spectroscopy, confirming isomeric and enantiomeric purity. The crystal structures of three of these complexes, [Pt(S meahaz)-Cl2], [Pt(R-etahaz)Cl2], and [Pt(S-dimeahaz)Cl2] (meahaz = N-methylahaz, etahaz = N-ethylahaz, dimeahaz = N,N-dimethylahaz), have been determined to establish the orientation of the protons and alkyl substituents on the nitrogen donor atoms. In vitro assays of the cytotoxic activity of the complexes have revealed a significant and reproducible enantioselective trend with the R enantiomers more active than the S-enantiomers in all cell lines. Increasing the steric bulk on the amine groups was found to have only a modest effect on activity. No enantioselectivity was observed in the binding of R- and S [Pt(etahaz)Cl2] to calf-thymus DNA. PMID- 9357518 TI - Novel tacrine analogues for potential use against Alzheimer's disease: potent and selective acetylcholinesterase inhibitors and 5-HT uptake inhibitors. AB - Several novel analogues of tacrine have been synthesized and tested for their ability to inhibit acetylcholinesterase, butyrylcholinesterase, and neuronal uptake of 5-HT (serotonin) and noradrenaline. Changes in the size of the carbocyclic ring of tacrine produced modest potency against cholinesterase enzymes. Addition of a fourth ring resulted in compounds with marked selectivity for acetylcholinesterase (AChE) over butyrylcholinesterase (BChE): e.g. 6-amino 4,5-benzo-5H-cyclopenta[1,2-b]-quinoline (14a) had an IC50 of 0.35 microM against AChE and 3.1 microM against BChE. Some tetracyclic compounds are 100-400 times more active than tacrine as inhibitors of neuronal uptake of serotonin, in particular 13-amino-6,7-dihydro-5H-benzo-[3,4]cyclohepta[1,2-b]quinoline (18), which had an IC50 of 20 nM. These compounds would be expected to facilitate both cholinergic and monoaminergic transmission. They should be worth investigating in models of memory impairment. PMID- 9357519 TI - Inhibition of tubulin polymerization by 5,6-dihydroindolo[2,1-alpha]isoquinoline derivatives. AB - 6-Alkyl-12-formyl-5,6-dihydroindolo[2,1-alpha]isoquinolines have been shown to inhibit the growth of human mammary carcinoma cells by an unknown mode of action. One of the possible molecular targets is the tubulin system which is involved in cell division. A number of 5,6-dihydroindolo[2,1-alpha]isoquinolines with methoxy or hydroxy groups in positions 3, 9, and/or 10 and various functional groups such as formyl, acetyl, cyano, alkylimino, and alkylamino in position 12 were synthesized and evaluated for both inhibition of tubulin polymerization and cytostatic activity in MDA-MB 231 and MCF-7 human breast cancer cells. In the tubulin polymerization assay, only hydroxy derivatives were active, whereas both the hydroxy derivatives and some of the methoxy compounds inhibited cell growth. In order to establish a correlation between the inhibition of tubulin polymerization and cytostatic activity in the hydroxy series, two of the most active racemates were separated into the enantiomers. In both assays, the relative potencies of the hydroxy derivatives were in a similar order. Highest activity was found for the (+)-isomers of 6-propyl- (6b) and 6-butyl-12-formyl 5,6-hydro-3,9-dihydroxyindolo[2,1-alpha]isoquino line (6c) with IC50 values of 11 +/- 0.4 and 3.1 +/- 0.4 microM, respectively, for the polymerization of tubulin at 37 degrees C (colchicine: 2.1 +/- 0.1 microM). The active hydroxy derivatives displaced 40-70% of [3H]colchicine from its binding site in the tubulin at concentrations 10-fold higher than that of colchicine. The data suggest that hydroxy-substituted indolo[2,1-alpha]isoquinolines bind to the colchicine-binding site and inhibit the polymerization of tubulin. This action can be assumed to be responsible for the cytostatic activity of the hydroxy derivatives and might also contribute to the antitumor effect of the corresponding methyl ethers. PMID- 9357520 TI - Studies on selection blockers. 5. Design, synthesis, and biological profile of sialyl Lewis x mimetics based on modified serine-glutamic acid dipeptides. AB - We have rationally designed a sLe(x) mimetic based on molecular modeling, synthesized type II and type II' beta-turn dipeptides (3a,b), and evaluated their biological profiles both in vitro and in vivo. Against E-selectin-sLe(x) binding, the type II beta-turn dipeptide L-Ser-D-Glu 3a (IC50, 13 microM) and the type II' beta-turn dipeptide D-Ser-L-Glu 3b (IC50, 5.5 microM) were 20-100-fold more potent blockers than sLe(x) (1; IC50, 600 microM) and a 3'-sulfated Le(x) analog (2; IC50, 280 microM). On the other hand, other stereoisomers, such as L-Ser-L Glu 3c and D-Ser-D-Glu 3d, were very weak blockers, with IC50 > 1000 microM for both 3c,d. Against the P- and L-selectins, despite much different stereochemistry of compounds 3a-d, the dipeptides 3a-d were all more potent blockers than either sLe(x) or compound 2. Interestingly, compound 3b provided significant in vivo efficacy against an immunoglobulin E-mediated skin reaction in a mouse model. These findings indicate that sLe(x) mimetics with type II and type II' beta-turn dipeptides could be useful in the design of an active selectin blocker in vitro and/or in vivo. PMID- 9357522 TI - Discovery of 3-aminobenzyloxycarbonyl as an N-terminal group conferring high affinity to the minimal phosphopeptide sequence recognized by the Grb2-SH2 domain. AB - The observation that anthranilic acid as N-terminal group produces a dramatic increase of the binding affinity of the phosphopeptide sequence Glu-pTyr-Ile-Asn for the Grb2-SH2 domain was rationalized by molecular modeling. The model, which invokes a stacking interaction between the N-terminal group and the SH2 domain residue Arg alpha A2, was subsequently used to design the 3 aminobenzyloxycarbonyl N-terminal group. The latter confers high affinity (IC50 = 65 nM in an ELISA assay) to the minimal sequence pTyr-Ile-Asn recognized by the Grb2-SH2 domain. PMID- 9357521 TI - Differentiation between partial agonists and neutral 5-HT1B antagonists by chemical modulation of 3-[3-(N,N-dimethylamino)propyl]-4-hydroxy- N-[4-(pyridin-4 yl)phenyl]benzamide (GR-55562). AB - The synthesis and binding affinity at cloned h5-HT1D, h5-HT1D, and h5-HT1A receptors of 3-[3-(N,N-dimethylamino)propyl]-4-hydroxy- N-[4-(pyridin-4 yl)phenyl]benzamide (2, GR-55562) and four O-methylated analogs are described. The functional activity of these compounds was determined at the h5-HT1B receptor using a [35S]GTP gamma S binding assay. The four analogs have been prepared in order to evaluate the influence of the alkylamino side chain conformation on binding and intrinsic activity. Whereas 2 and its derivatives display a similar binding affinity profile, major differences arise from analysis of the intrinsic activity data at h5-HT1B receptors. The O-methylated analog of 2, 3-[3-(N,N dimethylamino)propyl]-4-methoxy- N-[4-(pyridin-4-yl)phenyl]benzamide (3a), and the (1Z)-3-(N,N-dimethylamino)prop-1-enyl derivative (3c) act as neutral and potent antagonists (in a similar way to 2), while the 3-(N,N-dimethylamino)-prop 1-ynyl (3b) and (1E)-3-(N,N-dimethylamino)prop-1-enyl (3d) analogs display nonnegligible agonist activity. Molecular modeling studies show that, when the common triaryl portions of the molecules are overlapped, the partial agonists and the neutral antagonists differ by a near-orthogonal orientation of the NH+ projection to the hydrogen-bond receptor site. PMID- 9357523 TI - Antimalarial activity of molecules interfering with Plasmodium falciparum phospholipid metabolism. Structure-activity relationship analysis. AB - A series of 80 compounds, primary, secondary, and tertiary amines and quaternary ammonium and bisammonium salts, most of them synthesized as potential choline or ethanolamine analogs, were tested against the in vitro growth of Plasmodium falciparum, the human malaria parasite. They were active over the 10(-3)-10(-8) M concentration range. A structure-activity relationship study was carried out using autocorrelation vectors as structural descriptors, and multidimensional analysis. Principal component analysis, ascending hierarchical classification, and stepwise discriminant analysis showed that both the size and shape of the molecule were essential for antimalarial potency, making the lipophilicity and electronegativity distribution in the molecular space essential. Using the autocorrelogram describing the molecular shape and the electronegativity distribution on the molecular graph, 98% of the molecules were correctly classified either as poorly active or active with only three explanatory variables. The most active compounds were quaternary ammoniums salts whose nitrogen atom had only one long lipophilic chain of 11 or 12 methylene groups (E5, E6, E10, E13, E20, E21, E22, E23, F4, F8), or the bisammoniums whose polar heads were linked by linear alkyl chains of 10 to 12 carbon atoms (G4, G23). The hydroxyethyl group of choline was not very beneficial, whereas the charge and substitutions of nitrogen (aimed at increasing lipophilicity) were essential for optimal interactions. A crude topographic model of the ligand (choline) binding site was thus drawn up. PMID- 9357524 TI - Biologically active heteroarotinoids exhibiting anticancer activity and decreased toxicity. AB - A series of retinoids, containing heteroatoms in a cyclic ring and called heteroarotinoids, were synthesized, and their biological activity was evaluated using tissue culture lines that have measurable responses to trans-retinoic acid (t-RA). Transglutaminase (TGase) was assessed in the human erythroleukemia cell line (GMO6141A) as an indicator of differentiation and apoptosis. Proliferation was evaluated in a human cervical cell line, CC-1, which exhibits dose-dependent alterations in growth rate in response to treatment with trans-retinoic acid. Activation of nuclear retinoic acid receptors was determined in a reporter cell line established from CC-1. The reporter line, called CC-B, contains a reporter gene controlled by a retinoic acid responsive element (RARE) and a thymidine kinase (tk) promoter. Treatment of the CC-B line with the heteroarotinoids resulted in a dose-responsive and retinoid-dependent regulation of reporter gene expression. The heteroarotinoids exhibited activity in all assays and correlated in a statistically significant manner between assays. RARE transactivation activity in CC-B cells correlated with induction of TGase in GMO6141A (R = 0.96) and with a decrease in the growth rate of CC-1 cells (R = -0.90). The ability of the selected heteroarotinoids to induce differentiation, inhibit proliferation, and activate nuclear receptors demonstrates the chemotherapeutic potential of these agents. In view of the biological activity cited, an in vivo toxicity study was conducted on male B6D2F1 mice with three heteroarotinoids, namely 8 [(2E,4E,6E)-3,7-dimethyl-7-(1,2,3,4-tetrahydro-4,4-dimeth ylthiochroman-6-yl) 2,4,6-heptatrienoic acid], 10 [(2E,4E,6E)-3,7-dimethyl-7-(1,2,3,4-tetrahydro-4,4 dimeth ylchroman-6-yl)-2, 4,6-heptatrienoic acid], and 13 [(E)-p-[2-(4,4 dimethylchroman-6-yl)propenyl]benzoic acid]. The mice were used with gavage of heteroarotinoids in corn oil [0.1, 0.2, 0.4, or 0.8 mg/kg] and with 0.01 or 0.05 mg/kg of TTNPB (5) [(E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2 naphthalenyl)-1- propenyl]benzoic acid] as reference controls. The target organs affected in the mice by the three heteroarotinoids were those typically associated with t-RA (1) toxicity. The maximum tolerated dose (MTD) of 13 was 9.4 mg/kg/day, which was equal in toxicity to that of t-RA (1) and 1000-fold less toxic than TTNPB (5). The MTDs of 8 and 10 were 34 and 32 mg/kg/day, respectively, which is 3-fold less toxic than t-RA (1) and 3000-fold less toxic than TTNPB (5). The 3000-fold reduced toxicity, compared with only a 27% reduction biological activity of 8 and 10 with respect to that of TTNPB, observed in our assays indicates a good therapeutic ratio of these heteroarotinoids over the parent compound. The biological activity and reduced toxicity of these heteroartinoids demonstrate the potential efficacy as anticancer agents. PMID- 9357525 TI - Novel fluoroquinolone antibacterial agents containing oxime-substituted (aminomethyl)pyrrolidines: synthesis and antibacterial activity of 7-(4 (aminomethyl)-3-(methoxyimino)pyrrolidin-1-yl)-1-cyclopropyl-6- fluoro-4-oxo-1,4 dihydro[1,8]naphthyridine-3-carboxylic acid (LB20304). AB - New pyrrolidine derivatives, which bear an alkyloxime substituent in the 4 position and an aminomethyl substituent in the 3-position of the pyrrolidine ring, have been synthesized and coupled with various quinolinecarboxylic acids to produce a series of new fluoroquinolone antibacterials. These fluoroquinolones were found to possess potent antimicrobial activity against both Gram-negative and Gram-positive organisms, including methicillin resistant Staphylococcus aureus (MRSA). Variations at the C-8 position of the quinolone nucleus included fluorine, chlorine, nitrogen, methoxy, and hydrogen atom substitution. The activity imparted to the substituted quinolone nucleus by the C-8 substituent was in the order F (C5-NH2) > F (C5-H) > naphthyridine > Cl = OMe = H against Gram positive organisms. In the case of Gram-negative strains, activity was in the order F (C5-NH2) > naphthyridine = F (C5-H) > H > Cl > OMe. The advantages provided by the newly introduced oxime group of the quinolones were clearly demonstrated by their comparison to a desoximino compound 30. In addition, the oxime moiety greatly improved the pharmacokinetic parameters of the novel quinolones. Among these compounds, compound 20 (LB20304) showed the best in vivo efficacy and pharmacokinetic profile in animals, as well as good physical properties. The MICs (microgram/mL) of LB20304, compound 30, and ciprofloxacin against several test organisms are as follows: S. aureus 6538p (0.008, 0.031, and 0.13), methicillin resistant S. aureus 241 (4, 16, and 128), Streptococcus epidermidis 887E (0.008, 0.016, and 0.13), methicillin resistant S. epidermidis 178 (4, 32, and 128), Enterococcus faecalis 29212 (0.063, 0.13, and 1), Pseudomonas aeruginosa 1912E (0.25, 0.5, and 0.13), Escherichia coli 3190Y (0.008, 0.016, and 0.008), Enterobacter cloacae P99 (0.008, 0.031, and 0.008), Actinobacter calcoaceticus 15473 (0.063, 0.13, and 0.25). On the basis of these promising results, LB20304 was selected as a candidate for further evaluation. PMID- 9357526 TI - Design, synthesis, and dopamine receptor modulating activity of diketopiperazine peptidomimetics of L-prolyl-L-leucylglycinamide. AB - The diketopiperazine "C5" conformational mimic has been incorporated into the L prolyl-L-leucylglycinamide (PLG, 1) structure and into the bicyclic lactam PLG peptidomimetic structure 3 to give compounds 5 and 6, respectively. These analogues were designed to explore the idea that the N-terminal "C5" conformation, which was found in the crystal structure of 2 and which was mimicked in 4 by the diketopiperazine function, was a factor in the high potency of these two agents. Through the use of the [3H]spiroperidol/N propylnorapomorphine (NPA) D2 receptor competitive binding assay, both 5 and 6 were found to increase the affinity of the dopamine receptor for agonists and both were found to increase the percentage of D2 receptors which existed in the high-affinity state. These effects were observed when Gpp(NH)p was either absent or present, and they were analogous to the effects observed previously for PLG and the PLG peptidomimetics 2 and 4. However, the potency seen with 5 and 6 was less than that seen for 2 and 4, suggesting that while the N-terminal "C5" conformation may play a role in the potency of the gamma-lactam peptidomimetics of PLG, it does not appear to be the primary factor. In the 6-hydroxydopamine lesioned animal model of Parkinson's disease, 5 altered apomorphine-induced rotational behavior in a dose-dependent manner. The maximum effect occurred at a dose of 0.01 mg/kg i.p. and resulted in a 52.27 +/- 13.96% (p < 0.001, n = 7) increase in rotations compared to apomorphine administered alone. PMID- 9357527 TI - Use of a pharmacophore model for the design of EGF-R tyrosine kinase inhibitors: 4-(phenylamino)pyrazolo[3,4-d]pyrimidines. AB - In the course of the random screening of a pool of CIBA chemicals, the two pyrazolopyrimidines 1 and 2 have been identified as fairly potent inhibitors of the EGF-R tyrosine kinase. Using a pharmacophore model for ATP-competitive inhibitors interacting with the active site of the EGF-R protein tyrosine kinase (PTK), the class of the pyrazolo[3,4-d]pyrimidines was then optimized in an interactive process leading to a series of 4-(phenylamino)-1H-pyrazolo[3,4-d] pyrimidines as highly potent inhibitors of the EGF-R tyrosine kinase. The most potent compounds 13, 14, 15, 17, 19, 22, 26, 28, and 30 of this series inhibited the EGF-R PTK with IC50 values below 10 nM. High selectivity toward a panel of nonreceptor tyrosine kinases (c-Src, v-Abl and serine/threonine kinases (PKC alpha, CDK1) was observed. In cells, EGF-stimulated cellular tyrosine phosphorylation was inhibited by compounds 13, 15, 19, 22, and 23 at IC50 values below 50 nM, whereas PDGF-induced tyrosine phosphorylation was not affected by concentrations up to 10 microM, thus indicating high selectivity for the inhibition of the ligand-activated EGF-R signal transduction pathway. Compounds 15 and 19 inhibited proliferation of the EGF-dependent MK cell line with IC50 values below 0.5 microM. In addition, two compounds, 9 and 11, showing satisfactory oral bioavailability in mice after oral administration, exhibited good in vivo efficacy at doses of 12.5 and 50 mg/kg in a nude mouse tumor model using xenografts of the EGF-R overexpressing A431 cell line. From SAR studies, a binding mode for 4-(phenylamino)-1H-pyrazolo[3,4-d]pyrimidines, especially for compound 15, at the ATP-binding site of the EGF-R tyrosine kinase is proposed. 4 (Phenylamino)-1H-pyrazolo[3,4-d]pyrimidines represent a new class of highly potent tyrosine kinase inhibitors which preferentially inhibit the EGF-mediated signal transduction pathway and have the potential for further evaluation as anticancer agents. PMID- 9357528 TI - Potent anandamide analogs: the effect of changing the length and branching of the end pentyl chain. AB - To examine the effect of changing the length and branching of the end pentyl chain (C5H11) of anandamide (AN), various analogs 1a-h and 2a-f were synthesized from either the known aldehyde ester 6a or from the alcohol 6b and tested for their pharmacological activity. A reproducible procedure was developed for the conversion of arachidonic acid to 6a or 6b in gram quantities (overall yield 15%). The appropriate tetraene esters 7 were prepared by carrying out a Witting reaction, between 6a and the ylide generated from the phosphonium salt of the appropriate alkyl halide or between the ylide of 6d (prepared from 6a-->6b-->6c- >6d) and the appropriate alkyl aldehydes. They were then hydrolyzed to the corresponding acids and transformed into AN analogs 1 via their acid chlorides then treated with excess ethanolamine. alpha-Alkylation of esters 7 gave compounds 8 which were hydrolyzed to the corresponding acids. These acids via their acid chlorides and subsequent treatment with excess fluoroethylamine gave the target compounds 2. In this way analogs 1e and 2a-c were synthesized from 6d while all the remaining analogs were prepared from 6a. In order to assess the optimal length of the alkyl terminus, analogs 1a-d were prepared and showed moderately high affinities (18-55 nM). However analogs 1a-c failed to produce significant pharmacological effects at doses up to 30 mg/kg. Analog 1d was found to be a weak partial agonist. The reason for the lack of activity in 1a-c is presently not clear. Like the THCs, the branching of the end pentyl chain in AN (1e-h) increased potency both in in vitro and in vivo activities; the dimethylheptyl (DMH) analog 1e was the most potent in the series. Similar alkyl substitutions were carried out in the fluoro-2-methylanandamide series (2a-f), and all of these analogs had high receptor affinities (1-14 nM), the DMH analog 2a being the most potent. With a few exceptions they showed robust pharmacological effects, and AN-like profiles. It was shown that the SAR of the end pentyl chain in AN is very similar to that of THCs. However, the magnitude of enhanced potency observed when the side chain of THC was changed from straight to branched was not observed when the end chain of AN was similarly changed. PMID- 9357530 TI - Structure-activity relationships of 2'-deoxy-2',2'-difluoro-L-erythro pentofuranosyl nucleosides. AB - Following the recent discoveries that some L-nucleosides are more or equal potent than their D-counterparts, we synthesized 2'-deoxy-2',2'-difluoro-L-erythro pentofuranosyl nucleosides as potential antiviral agents. The target compounds were synthesized via the key intermediates 7a or 7b from L-gulono gamma-lactone. Compound 2 was oxidatively cleaved and coupled with ethyl bromodifluoroacetate in the presence of activated zinc under Reformatsky conditions to obtain a diasteomeric mixture of 4(R) and 4(S), in a 4:1 ratio. The major 4(R) isomer was cyclized and treated appropriately to obtain the mesylate 8a or 8b, which was condensed with various silyl-protected pyrimidines. Condensation of the alcohol 7a or 7b with 6-chloropurine under Mitsunobu conditions afforded the 6 chlorpurine analogs 53a or 53b and 54a or 54b. Further treatment of the compounds 53a, 54a and 53b, 54b afforded the inosine and adenine derivatives 57-60, respectively. The condensation of 2-amino-6-chloropurine with compound 8a and subsequent treatment with 2-mercaptoethanol/sodium methoxide afforded the guanine analogs 63 and 64. All of the synthesized nucleosides 31-52, 57-60, 63, and 64 were evaluated for antiviral activity and for cellular toxicity. Adenine derivative 57 showed a moderate activity against HIV-1 in PBM cells (3.4 microM). None of the other compounds showed any significant activities against HIV-1, HBV, HSV-1, HSV-2, and toxicity in Vero, CEM, and PBM cell lines up to 100 microM. The X-ray structure of the 5-iodocytosine analog showed a 2'-exo/3'-endo conformation for the carbohydrate moiety, which is different from those of the biologically active compounds (-)-FTC and L-FMAU. PMID- 9357529 TI - Synthesis and pharmacological comparison of dimethylheptyl and pentyl analogs of anandamide. AB - (Dimethylheptyl)anandamide [(16,16-dimethyldocosa-cis-5,8,11,14 tetraenoyl)ethanolamine ] (17a) and its amide analogs were synthesized by Wittig coupling of a ylide derived from a fragment of arachidonic acid. These amides were compared to the endogenous cannabinoid receptor ligand arachidonylethanolamide (anandamide, 2a) and its amide analogs in pharmacological assays for potential enhancement of cannabimimetic activities. The receptor affinity to rat brain membranes of the dimethylheptyl (DMH) analogs increased by an order of magnitude in most comparisons to the corresponding anandamides in displacement assays versus the cannabinoid agonist [3H]CP 55,940 or antagonist [3H]SR141716A, for which rank order differences in affinity were observed. An order of magnitude enhancement of potency with comparable or higher efficacy in behavioral assays in the mouse tetrad of tests of cannabinoid activity was observed in 17a versus 2a. In contrast, no enhancement in potency for the pentyl to DMH side chain exchange was seen in the mouse vas deferens assay. The data indicate a structural equivalence between classical plant cannabinoids and 2a as well as different receptor-ligand interactions that characterize multiple receptor sites or binding modes. PMID- 9357531 TI - Synthesis of willardiine and 6-azawillardiine analogs: pharmacological characterization on cloned homomeric human AMPA and kainate receptor subtypes. AB - Both willardiine and azawillardiine analogs (18-28) have been reported to be potent and selective agonists for either AMPA or kainate receptors. We report here the novel synthesis and pharmacological characterization of a range of willardiine (18-23) and 6-azawillardiine (24-28) analogs on cells individually expressing human homomeric hGluR1, hGluR2, hGluR4, or hGluR5 receptors. Reaction of the sodium salts of substituted uracils (7-12) or 6-azauracils (13-16) with (S)-3-[(tert-butoxycarbonyl)amino]oxetan-2-one (17) in dry DMF, subsequent deprotection in TFA, and purification by ion-exchange chromatography gave mainly the willardiine analog in which alkylation took place on N1 of the uracil ring. We have investigated the subtype selectivity of these compounds by examining their binding affinity for homomeric hGluR1, -2, -4, or -5 (and hGluR6 in the case of 5-iodowillardiine (22)). From this study we have demonstrated that 22 has high affinity for hGluR5 and, compared to kainate, displays excellent selectivity for this receptor over both the AMPA receptor subtypes and the homomeric kainate receptor, hGluR6. 5-Fluorowillardiine (19) has higher affinity than AMPA for both homomeric hGluR1 and hGluR2 and compared to AMPA displays greater selectivity for AMPA receptor subtypes over the kainate receptor, hGluR5. Some structural features required for optimal activity at homomeric AMPA or kainate receptor subtypes have also been identified. It would appear that quite large lipophilic substituents at the 5-position of the uracil ring not only are accommodated by hGluR5 receptors but also lead to enhanced affinity for these receptors. In contrast to this, for optimal binding affinity to hGluR1, -2, or -4, smaller, electron-withdrawing substituents are required. For optimal activity at hGluR4 receptors a 6-aza-substituted willardiine is favored. The subtype-selective compounds described here are likely to be useful tools to probe the distribution and the physiological roles of the various glutamate receptor subunits in the central nervous system. PMID- 9357532 TI - Constrained corticotropin-releasing factor antagonists with i-(i + 3) Glu-Lys bridges. AB - Hypothesis driven and systematic structure-activity relationship (SAR) investigations have resulted in the development of effective central nervous system (CNS) antagonists of corticotropin (ACTH)-releasing factor (CRF) such as alpha-helical CRF(9-41) and analogues of our assay standard [DPhe12,Nle21,38]hCRF(12-41). On the other hand, equally potent CRF antagonists that block the hypothalamic/pituitary/adrenal (HPA) axis had not been described until recently. Predictive methods, physicochemical measurements (nuclear magnetic resonance spectrometry and circular dichroism spectroscopy), and SAR studies suggest that CRF and its family members (urotensins and sauvagine) assume an alpha-helical conformation when interacting with CRF receptors. To further test this hypothesis, we have systematically scanned the hCRF(9-41) or hCRF(12 41) sequences with an i-(i + 3) bridge consisting of the Glu-Xaa-Xbb-Lys scaffold which we and others had shown could maintain or enhance alpha-helical structure. From this series we have identified seven analogues that are either equipotent to, or 3 times more potent than, the assay standard; in addition, as presented earlier cyclo(30-33)[DPhe12,-Nle21,38,Glu30, Lys33]hCRF(12-41) (astressin) is 32 times more potent than the assay standard in blocking ACTH secretion in vitro (rat pituitary cell culture assay). In vivo, astressin is also significantly more potent than earlier antagonists at reducing hypophysial ACTH secretion in intact stressed or adrenalectomized rats. Since the corresponding linear analogues that were tested are significantly less potent, our interpretation of the increased potency of the cyclic analogues is that the introduction of the side chain to side chain bridging element (Glu30-Lys33, and to a lesser extent that of Glu14 Lys17, Glu20-Lys23, Glu23-Lys26, Glu26-Lys29, Glu28-Lys31, Glu29-Lys32, and Glu33 Lys36) induces and stabilizes in the receptor environment a putative alpha helical bioactive conformation of the fragment that is not otherwise heavily represented. The effect of the introduction of two favored substitutions [(cyclo(20-23) and cyclo(30-33)] yielded 37 with a potency 8 times that of the assay standard but actually 12 times less than expected if the effect of the two cycles had been multiplicative. These results suggest that the pituitary CRF receptor can discriminate between slightly different identifiable conformations, dramatically illustrating the role that secondary and tertiary structures play in modulating biological signaling through specific protein-ligand interactions. PMID- 9357533 TI - Induction of the estrogen specific mitogenic response of MCF-7 cells by selected analogues of estradiol-17 beta: a 3D QSAR study. AB - Analogues of estradiol-17 beta (E2) have been evaluated for estrogen receptor (ER) binding affinity and mitogenic potential in the human breast cancer cell line MCF-7. These 42 compounds represent subtle modifications of the natural estrogen structure through the placement of hydroxyl, amino, nitro, or iodo groups around the ring system in addition to, or as replacement of, the 3- and 17 beta-hydroxyls of E2. The mitogenic activity of the analogues was found to be related to ER binding only to a limited extent. In order to elucidate structural features that are uniquely responsible for receptor binding affinity or mitogen potential of estrogens, the three-dimensional quantitative structure-activity (QSAR) method Comparative Molecular Field Analysis (CoMFA) was employed. Separate CoMFA models for receptor binding and cell growth stimulation were optimized through the use of various alignment rules and region step size. Whereas the CoMFA contour plots did outline the shared structural requirements for the two measured biological properties, specific topological features in this set of estrogens were delineated that distinguish mitogenic potential from ER binding ability. In particular, steric interference zones which affected growth extend in a band from above the A-ring to position 4 and below, whereas the ER binding steric interference zones are limited to isolated polyhedra in the 1, 2 and 4 positions and the alpha face of the B-ring. In addition, electronegative features located around the A-, B-, or C-rings contribute to receptor affinity. However, growth is dependent only on electronegative and electropositive properties near the 3-position. In a final QSAR model for the mitogenic response, the value of ER binding was included along with structural features as a descriptor in CoMFA. The resulting 3D-QSAR has the most predictive potential of the models in this study and can be considered a prototype model for the general evaluation of a steroidal estrogen's growth stimulating ability in MCF-7 cells. For example, the location of D-ring contours illustrate the model's preference for 17 beta-hydroxy steroids over the less mitogenic 17 alpha- and 16 alpha-hydroxy compounds. In addition, the enhanced mitogenic effect of steric bulk in the 11 alpha-position is also evident. The QSAR studies in this report illustrate the fact that while ER binding may be a required factor of the estrogen dependent growth response in MCF 7 cells, particular structural characteristics, in addition to those responsible for tight receptor binding, must be present to induce an optimal mitogenic response. Therefore, this report demonstrates that the CoMFA QSAR method can be utilized to characterize structural features of test compounds that account for different types of estrogenic responses. PMID- 9357534 TI - Novel and highly potent 5-HT3 receptor agonists based on a pyrroloquinoxaline structure. AB - The synthesis and the biological evaluation of a series of novel pyrroloquinoxaline derivatives are described. In binding studies several compounds proved to be potent and selective 5-HT3 receptor ligands. The most active pyrroloquinoxalines, 11d and 11e, showed a subnanomolar affinity for 5-HT3 receptor and were able to functionally discriminate the central and peripheral 5 HT3 receptor, being agonists and antagonists, respectively. In functional studies ([14C]-guanidinium accumulation test in NG 108-15 cells, in vitro) most of the synthesized compounds showed clear-cut 5-HT3 agonist properties. In in vivo studies on the von Bezold-Jarisch reflex test (a peripheral interaction model) the behavior of the tested compounds ranged from agonist to antagonist, while clear agonist properties were obtained with 12a on cortical acetylcholine release in freely moving rats. Pharmacokinetic studies with 11e and 12c indicate that the compounds easily cross the blood-brain barrier (BBB) after systemic administration with a brain/plasma ratio of 17.5 and 37.5, respectively. Thus compounds 11e and 12c represent the most potent central 5-HT3 agonists identified to date that are able to cross the blood-brain barrier. PMID- 9357535 TI - 5-(N-oxyaza)-7-substituted-1,4-dihydroquinoxaline-2,3-diones: novel, systemically active and broad spectrum antagonists for NMDA/glycine, AMPA, and kainate receptors. AB - A group of 5-aza-7-substituted-1,4-dihydroquinoxaline-2,3-diones (QXs) and the corresponding 5-(N-oxyaza)-7-substituted QXs were prepared and evaluated as antagonists of ionotropic glutamate receptors. The in vitro potency of these QXs was determined by inhibition of [3H]-5,7-dichlorokynurenic acid ([3H]DCKA) binding to N-methyl-D-aspartate (NMDA)/glycine receptors, [3H]-(S)-alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid ([3H]AMPA) binding to AMPA receptors, and [3H]kainate ([3H]KA) binding to KA receptors in rat brain membranes. 5-(N Oxyaza)-QXs 12a-e all have low micromolar or submicromolar potency for NMDA/glycine receptors and low micromolar potencies for AMPA and KA receptors. QXs 12a-e display 2-12-fold selectivity for NMDA/glycine receptors compared to AMPA receptors, and approximately 2-fold difference between AMPA and KA potency. In contrast to other QXs that either show high selectivity for NMDA (such as ACEA 1021) or AMPA (such as NBQX) receptors, these molecules are broad spectrum antagonists of ionotropic glutamate receptors. 7-Nitro-5-(N-oxyaza)-QX (12e) is the most potent inhibitor among 12a-e, having IC50 values of 0.69, 1.3, and 2.4 microM at NMDA, AMPA, and KA receptors, respectively. In functional assays on glutamate receptors expressed in oocytes by rat cerebral cortex poly(A+) RNA, 7 chloro-5-(N-oxyaza)-QX (12a) and 7-nitro-5-(N-oxyaza)-QX (12e) have Kb values of 0.63 and 0.31 microM for NMDA/glycine receptors, and are 6- and 4-fold selective for NMDA over AMPA receptors, respectively. 5-(N-Oxyaza)-7-substituted-QXs 12a-e all have surprisingly high in vivo potency as anticonvulsants in a mouse maximal electroshock-induced seizure (MES) model. 7-Chloro-5-(N-oxyaza)-QX (12a), 7-bromo 5-(N-oxyaza)-QX (12b), and 7-methyl-5-(N-oxyaza)-QX (12c) have ED50 values of 0.82, 0.87, and 0.97 mg/kg i.v., respectively. The high in vivo potency of QXs 12a-e is particularly surprising given their low log P values (approximately 2.7). Separate studies indicate that QXs 12a and 12e are also active in vivo as neuroprotectants and also have antinociceptive activity in animal pain models. In terms of in vivo activity, these 5-(N-oxyaza)-7-substituted-QXs are among the most potent broad spectrum ionotropic glutamate antagonists reported. PMID- 9357536 TI - Synthesis of a series of potent and orally bioavailable thrombin inhibitors that utilize 3,3-disubstituted propionic acid derivatives in the P3 position. AB - As part of an effort to prepare efficacious and orally bioavailable analogs of the previously reported thrombin inhibitors 1a, b, we have synthesized a series of compounds that utilize 3,3-disubstituted propionic acid derivatives as P3 ligands. By removing the N-terminal amino group, the general oral bioavailability of this class of compounds was enhanced without excessively increasing the lipophilicity of the compounds. The overall properties of the molecules could be drastically altered depending on the nature of the groups substituted onto the 3 position of the P3 propionic acid moiety. A number of the compounds exhibited good oral bioavailability in rats and dogs, and numerous compounds were efficacious in a rat FeCl3-induced model of arterial thrombosis. Compound 7, the 3,3-diphenylpropionic acid derivative, showed the best overall profile of in vivo and in vitro activity. Molecular modeling studies suggest that these compounds bind in the thrombin active site in a manner essentially identical to that previously reported for compound 1a. PMID- 9357537 TI - 2,4-Diaminothieno[2,3-d]pyrimidine lipophilic antifolates as inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase. AB - Ten previously unreported 2,4-diaminothieno[2,3-d]pyrimidine lipophilic dihydrofolate reductase inhibitors were synthesized as potential inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase. Pivaloylation of 2,4-diamino-5-methylthieno[2,3-d]pyrimidine followed by dibromination with N bromosuccinimide in the presence of benzoyl peroxide gave 2,4-bis(pivaloylamino) 6-bromo-5-(bromomethyl)thieno[2,3-d]pyrimid ine, which after condensation with substituted anilines or N-methylanilines and deprotection with base yielded 2,4 diamino-6-bromo-5-[(substituted anilino)methyl]thieno[2,3-d]pyrimidines. Removal of the 6-bromo substituent was accomplished with sodium borohydride and palladium chloride. The reaction yields were generally good to excellent. The products were tested as inhibitors of dihydrofolate reductase (DHFR) from P. carinii, T. gondii, and rat liver. Although the IC50 could not be reached for the 6 unsubstituted compounds because of their extremely poor solubility, three of the five 6-bromo derivatives were soluble enough to allow the IC50 to be determined against all three enzymes. 2,4-Diamino-5-[3,5-dichloro-4-(1 pyrrolo)anilino]methyl]- 6-bromothieno[2,3-d]pyrimidine was the most active of the 6-bromo derivatives, with an IC50 of 7.5 microM against P. carinii DHFR, but showed no selectivity for either P. carinii or T. gondii DHFR relative to the enzyme from rat liver. PMID- 9357538 TI - (S)-homo-AMPA, a specific agonist at the mGlu6 subtype of metabotropic glutamic acid receptors. AB - Our previous publication (J. Med. Chem. 1996, 39, 3188-3194) described (RS)-2 amino-4-(3-hydroxy-5-methylisoxazol-4-yl)butyric acid (Homo-AMPA) as a highly selective agonist at the mGlu6 subtype of metabotropic excitatory amino acid (EAA) receptors. Homo-AMPA has already become a standard agonist for the pharmacological characterization of mGlu6 (Trends Pharmacol. Sci. Suppl. 1997, 37 39), and we here report the resolution, configurational assignment, and pharmacology of (S)- (6) and (R)- (7) Homo-AMPA. Using the "Ugi four-component condensation", 3-(3-ethoxy-5-methylisoxazol-4-yl)propanal (10) was converted into the separable diastereomeric derivatives of 6 and 7, compounds 12 and 11, respectively. Deprotection of 12, in one or two steps, gave extensively racemized 6, which was converted in low yield into 6 (99.0% ee) through several crystallizations. 6 (99.7% ee) and 7 (99.9% ee) were finally obtained by preparative chiral HPLC. The configurational assignments of 6 and 7 were based on 1H NMR spectroscopic studies on 12 and 11, respectively, and circular dichroism studies on 6 and 7. Values of optical rotations using different solvents and the chiral HPLC elution order of 6 and 7 supported the results of the spectroscopic configurational assignments. The activities of 6 and 7 at ionotropic EAA (iGlu) receptors and at mGlu1-7 were studied. (S)-Homo-AMPA (6) was shown to be a specific agonist at mGlu6 (EC50 = 58 +/- 11 microM) comparable in potency with the endogenous mGlu agonist (S)-glutamic acid (EC50 = 20 +/- 3 microM). Although Homo-AMPA did not show significant effects at iGlu receptors, (R)-Homo-AMPA (7), which was inactive at mGlu1-7, turned out to be a weak N-methyl-D-aspartic acid (NMDA) receptor antagonist (IC50 = 131 +/- 18 microM). PMID- 9357539 TI - Variable aberrant cDNAs in single diphtheria toxin-resistant human fibroblasts. AB - We treated transformed human fibroblasts with diphtheria toxin (DT) and isolated 40 single cells that were toxin resistant but unable to propagate. In 13 of them toxin resistance was associated with the presence of one or more aberrant transcripts of the structural gene for elongation factor 2 (EF-2). cDNA obtained from these transcripts had 164-447 bp-long deletions. Each of these deletions was associated with 2-8 base pairs-long repeats at its breakpoints. Only 10 out of 16 cDNA deletions were associated with presumed exon junctions. A role is suggested for errors in transcription in producing the aberrant transcripts which gave rise to the deletion-bearing cDNA species. PMID- 9357540 TI - Effects of dNTPs on the SCE frequency in plant root tip cells. AB - The effects of deoxynucleoside triphosphate (dNTPs) on the frequency of sister chromatid exchange (SCE) have been studied in plant root tip cells. Treatment with dATP caused a dose-dependent decrease in SCE frequency, while dGTP, caused a dose-dependent increase in SCE frequency both in Hordeum vulgare and in Vicia faba root tip cells. Treatment with dCTP and dTTP significantly increased SCE frequency in H. vulgare root tip cells, but significantly decreased SCE frequency in V. faba root tip cells. The SCE induced by treatment with dGTP could be totally reversed by treatment with an equal concentration of any of the other three dNTPs (dATP, dCTP and dTTP) both in H. vulgare and in V. faba. Treatment with a mixed equal concentration of dATP, dCTP, dGTP and dTTP did not alter the SCE frequency. These results suggest that dNTP pool imbalance is an SCE-affecting factor in plant cells. PMID- 9357541 TI - Increased level of chromosomal aberrations in lymphocytes of Chernobyl liquidators 6-10 years after the accident. AB - Chromosomal aberrations (CA) were used to investigate the level of cytogenetical damage in peripheral blood lymphocytes from the liquidators in a remote period (6 10 years) after the Chernobyl accident. There was a significantly higher frequency of chromosomal radiation markers (dicentrics and rings) in the peripheral lymphocytes of the liquidators than in the control subjects. No differences between these groups were demonstrated by the micronucleus (MN) test. Increased frequency of chromatid exchanges was associated with the smoking habits of the liquidators. PMID- 9357542 TI - X-ray-induced synaptonemal complex damage during meiotic prophase in female fetuses of Rattus norvegicus. AB - The different types of damage (synaptic anomalies, chromosome reorganizations and nucleolar fragmentation) observed in oocytes from female rat fetuses irradiated at 14, 16 and 18 days of gestation (d.g.), respectively, indicates the existence of a special sensitivity at the different stages of prophase to X-rays. At the highest dose (5 Gy), we observed a decrease in some of the synaptonemal complex indicators of chromosomal damage, probably reflecting a selection against cells with the highest degree of chromosome anomalies. PMID- 9357543 TI - Genotoxic effects of structurally related beta-carboline alkaloids. AB - beta-Carboline alkaloids, found in medicinal plants, tobacco smoke and well cooked foods, have shown a variety of actions in biological systems related to their interaction with DNA. Therefore, these alkaloids can be considered potentially mutagenic. In this work, the genotoxic, mutagenic, and cytotoxic activities of three aromatic beta-carboline alkaloids (harman, harmine, and harmol) and two dihydro-beta-carboline alkaloids (harmaline and harmalol) were evaluated by means of the Salmonella/microsome assay (Salmonella typhimurium TA98, TA97, TA100, and TA102) and SOS chromotest (Escherichia coli PQ37) with and without metabolic activation. Moreover, harman and harmine were analyzed by the micronucleus assay in vivo. It was shown that genotoxicity was inhibited by the addition of S9 mix for aromatic beta-carbolines harman and harmol in TA97. However, harmine showed signs of mutagenicity only in the presence of S9 mix in TA98 and TA97 frameshift strains. In the SOS chromotest, only harman induced SOS functions in the absence of S9 mix. Dihydro-beta-carbolines were not genotoxic in any of the microorganisms used. The negative responses obtained in the micronucleus assay indicated that harman and harmine were not able to induce chromosomal mutations. PMID- 9357544 TI - Chromosomal mutations and chromosome loss measured in a new human-hamster hybrid cell line, ALC: studies with colcemid, ultraviolet irradiation, and 137Cs gamma rays. AB - Small mutations, megabase deletions, and aneuploidy are involved in carcinogenesis and genetic defects, so it is important to be able to quantify these mutations and understand mechanisms of their creation. We have previously quantified a spectrum of mutations, including megabase deletions, in human chromosome 11, the sole human chromosome in a hamster-human hybrid cell line AL. S1- mutants have lost expression of a human cell surface antigen, S1, which is encoded by the M1C1 gene at 11p13 so that mutants can be detected via a complement-mediated cytotoxicity assay in which S1+ cells are killed and S1- cells survive. But loss of genes located on the tip of the short arm of 11 (11p15.5) is lethal to the AL hybrid, so that mutants that have lost the entire chromosome 11 die and escape detection. To circumvent this, we fused AL with Chinese hamster ovary (CHO) cells to produce a new hybrid, ALC, in which the requirement for maintaining 11p15.5 is relieved, allowing us to detect mutations events involving loss of 11p15.5. We evaluated the usefulness of this hybrid by conducting mutagenesis studies with colcemid, 137Cs gamma-radiation and UV 254 nm light. Colcemid induced 1000 more S1- mutants per unit dose in ALC than in AL; the increase for UV 254 nm light was only two-fold; and the increase for 137Cs gamma-rays was 12-fold. The increase in S1- mutant fraction in ALC cells treated with colcemid and 137Cs gamma-rays were largely due to chromosome loss and 11p deletions often containing a breakpoint within the centromeric region. PMID- 9357545 TI - QSAR treatment of multiple toxicities: the mutagenicity and cytotoxicity of quinolines. AB - A series of 15 quinoline congeners were assayed for mutagenicity and cytotoxicity in the Ames test using strain TA100 bacteria. Statistical analysis of the data allowed simultaneous determination of the mutagenicity and cytotoxicity of each quinoline. These data were used to develop three quantitative structure-activity relationships (QSAR). In all three QSAR, the strength of the relationship between hydrophobicity (as measured by log P) and biological activity was similar as h was near 1 in all three cases. For the mutagenicity of these quinolines, both hydrophobic and steric interactions appear to be important. In contrast, the cytotoxicity is mainly affected by increasing hydrophobicity and by the addition of electron withdrawing substituents to the quinoline ring. Comparison to other QSAR from our laboratory and others lends support to these findings. Both simultaneous consideration of different biological activities and the comparison of newly developed QSAR with previous data for the purpose of lateral validation should be encouraged in future QSAR studies. PMID- 9357546 TI - Normal telomere maintenance in immortal ataxia telangiectasia cell lines. AB - Telomeres are maintained in germ line cells and immortal cell lines, but shorten with each cell division in most somatic cells. Blood lymphocytes from individuals with ataxia telangiectasia (AT) demonstrate an accelerated rate of telomere shortening and high levels of telomere associations. This accelerated loss of telomeres in somatic cells in AT could be due to either the loss of more telomeric DNA with every cell division or an increased rate of cell division. The gene for AT shares homology with the yeast TEL1 gene, in which mutations result in abnormally shortened telomeres. Thus, mutations in the gene for ataxia telangiectasia may also influence the ability of germ line cells and immortal cell lines to properly maintain telomere homeostasis. To investigate a possible defect of telomere maintenance in AT we have analyzed 8 simian virus 40 (SV40) immortalized AT cell lines and twelve SV40-immortalized non-AT cell lines for both telomerase activity and telomere length. The results demonstrate that telomere length in AT cells is maintained via telomerase or an alternative (ALT) pathway in a manner indistinguishable from cell lines derived from normal cells. We also investigated telomere dynamics in one telomerase-positive AT cell line by analyzing the changes in the length of a single telomere, and found that this telomere maintained its equilibrium mean length (EML) similar to normal cell lines with stable chromosomes. The combined results show no significant differences between the telomeres of immortal AT and non-AT cell lines, demonstrating that the absence of wild-type ATM does not result in a fundamental defect in telomere maintenance in these cells. PMID- 9357547 TI - Spectrum of spontaneous and 2,4,6-trinitrotoluene (TNT)-induced mutations in Salmonella typhimurium strains with different nitroreductase and O acetyltransferase activities. AB - Spontaneous and 2,4,6-trinitrotoluene (TNT)-induced mutation spectra were determined at the hisD3052 allele of Salmonella typhimurium strains TA98, YG1021 (nitroreductase-overproducing) and YG1024 (O-acetyltransferase-overproducing). In TA98, 55% (11/20) of the spontaneous reversions and 95% (19/20) of reversions in TNT-treated plates were deletions of two bases at the same site (-2 hotspot deletions), whereas the respective figures were 65% (13/20) and 80% (16/20) in YG1021, and 75% (15/20) and 95% (19/20) in YG1024. Other mutations observed in the TNT treatment were complex frameshifts consisting of either a -2 hotspot deletion and a base substitution, or a +1 addition and base substitution at the stop codon. In addition, different kinds of deletions were recovered in the spontaneous spectra. The elevated enzymatic activities of strains, YG1021 and YG1024, resulting in enhanced mutagenicity of TNT, did not seem to have an effect on the spectrum of TNT-induced mutations. However, the YG strains, which also have a higher spontaneous revertant yield than the parental strain TA98, seemed to differ from TA98 in their spontaneous spectra. The increase consisted of revertants containing the -2 hotspot deletion, possibly indicating elevated activation of exogenous or endogenous premutagens by the higher enzyme activities of the YG strains. PMID- 9357549 TI - Reactive oxygen species-induced DNA damage and its modification: a chemical investigation. AB - The main purpose of this study was to determine whether well-known reactive oxygen species (ROS)-generating agents can induce DNA damage in a simple chemical system with or without Fenton reaction components (iron and reducing agents), and to explore whether antioxidants which normally exist in the cellular environment can modify such damage, i.e. to determine chemical reactions of relevance to biological systems. A neutral electrophoresis technique was used to investigate DNA double stranded breaks (DSBs) caused by chemical treatments of lambda-DNA in eppendorf tubes by various ROS-generating compounds and the degree of DNA damage was categorised by analysis of enhanced digital images. Double strand breaks were induced by hydroquinone (HQ), benzoquinone (BQ), benzenetriol (BT), hydrogen peroxide (H2O2), bleomycin (BLM) and sodium ascorbate (Vit C). DNA damage was modulated by various agents including catalase (CAT), superoxide dismutase (SOD), desferoxamine mesylate (DFO), ferrous chloride (FeCl2), reduced glutathione (GSH), trolox, silymarin and myricetin. Individual chemicals (except BLM) at the concentration of 1 mM did not induce large numbers of DSBs without iron [Fe(II) or Fe(III) at 25 microM]. GSH enhanced the damaging effect of HQ, BT and Vit C, did not alter the non-damaging effect of H2O2, but had a small protective effect on BLM. When compared with the non-enzyme protein, bovine serum albumin (BSA), SOD had a protective effect against BT, H2O2 and BLM; in the presence of GSH, SOD diminished the effect of HQ, BQ and Vit C but enhanced the effect of BT, H2O2 and BLM. With both GSH and Fe and compared with BSA, SOD enhanced the effect of HQ, BQ and BLM, ameliorated the effect of H2O2, and did not affect the others. CAT showed a protective effect for almost all examined compounds, but had little effect on BLM. With GSH alone, DFO enhanced the effect of HQ, BQ, H2O2 and ameliorated the effect of BT, BLM and Vit C and trolox was largely protective. With GSH and Fe, DFO was protective for all compounds except doxorubicin (Dox), trolox was protective for all compounds except Dox and BLM, silymarin was protective except that it had little effect on BLM and Dox, but myricetin did not show any protective effect. In conclusion, the results from the present study have further highlighted the adverse potential of reducing agents and redox cycling agents, and also the need for a cautious view of antioxidants. PMID- 9357548 TI - Characterization of a macromolecular matrix isolated from tobacco suspension cell cultures and its role in the activation of promutagenic m-phenylenediamine. AB - The medium recovered from the tobacco cell suspension cultures (TX1MX) activated the promutagenic aromatic amine m-phenylenediamine (mPDA) and a macromolecular complex (gel) responsible for the arylamine activation was isolated from the medium. The gel formation and the role of the gel components in the plant activation of mPDA to products mutagenic in S. typhimurium YG1024 were studied. The activation of mPDA was caused by the peroxidases present in TX1MX. We demonstrated an association of the peroxidase activity and gel pectins. Formation of a stable mutagenic association of mPDA with the macromolecular material was observed. The data indicate that the gel isolated from TX1MX is the macromolecular component of the arylamine conjugate proposed in earlier work. PMID- 9357550 TI - Nitric oxide as a carcinogen: analysis by yeast functional assay of inactivating p53 mutations induced by nitric oxide. AB - We have used a yeast p53 functional assay to study induction of mutations in the p53 tumor suppressor gene by nitric oxide and cytosine methylation. The yeast assay identifies only biologically important p53 mutations. p53 cDNA was treated with the nitric oxide donor sydnonimine, PCR-amplified and transfected into yeast. Sydnonimine produced a significant, dose-dependent increase in C:G-->A:T transversions. Many important p53 mutational hotspots are postulated to arise by deamination of methylCpG in tumors. We therefore examined nitric oxide induction of mutations in p53 cDNA methylated by PCR-mediated substitution of 5 methylcytosine for cytosine or by treatment with the SssI CpG methylase. Both methylation procedures increased the baseline mutation rate, and nitric oxide treatment produced a further increase in mutation yield. Sequence analysis showed that methylation alone led to C:G-->T:A transitions, whereas nitric oxide treatment simply produced more C:G-->A:T transversions. Thus the most important factor in C:G-->T:A transition at CpG sites identified in this experimental system is cytosine methylation, consistent with spontaneous conversion of 5 methylcytosine to thymine by deamination. PMID- 9357551 TI - Induction of specific-locus and dominant lethal mutations in male mice by 1,3 bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1 nitrosourea (CCNU). AB - 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1 nitrosourea (CCNU) induced dominant lethal and specific-locus mutations in male mice. For both compounds the germ cell stage sensitive to the induction of dominant lethal mutations was dose dependent. A dose of 5 mg BCNU per kg b.wt. induced dominant lethal mutations primarily in spermatocytes, whereas higher doses of BCNU induced dominant lethals in spermatids and spermatocytes. Following doses of 5 and 10 mg CCNU per kg b.wt. dominant lethals were induced in spermatids and spermatocytes similar to the results for higher doses of BCNU. Higher dose exposure to BCNU and CCNU was associated with dominant lethals expressed as pre-implantation loss (reduction in total number of implants). In addition, higher doses of CCNU showed a cytotoxic effect in differentiating spermatogonia. Both compounds induced specific-locus mutations in post spermatogonial germ cell stages of mice. However, CCNU increased also the specific-locus mutation frequency in spermatogonia in two out of three experiments. We conclude in analogy with criteria developed by IARC, that BCNU and CCNU are potential human mutagens. PMID- 9357552 TI - Computerized image analysis of sister chromatid exchanges in Chinese hamster ovary cells. AB - In this paper we show that sister chromatid exchanges (SCE) can be analysed by computer-supported determination of areas and shapes of darkly stained chromatid sections. In combination with appropriate statistical analyses, our method leads to measurements of SCE frequencies in neocarzinostatin-treated Chinese hamster ovary cells which are nearly as precise as the ones obtained by classical microscopical analyses. We discuss our data as an approach to the development of a fully automated SCE scoring system. PMID- 9357553 TI - Mutagenicity and cytotoxicity of reactive oxygen and nitrogen species in the MN 11 murine tumor cell line. AB - There is increasing evidence that endogenously generated reactive oxygen (ROS) and reactive nitrogen (RNS) species at sites of inflammation and in tumors may be genotoxic. We have developed a murine tumor model (MN-11) in which mutations at the hypoxanthine phosphoribosyltransferase (HPRT) locus, arising both in vitro and in vivo, can be detected. In the present report, we describe an in vitro study of the ability of ROS and RNS to induce mutations in our model system. 137Cs radiation and radiomimetic drugs caused a dose-dependent increase in mutant frequency. At D0, radiation induced about 170 mutants per 10(5) viable cells, compared to 50 and 95 for streptonigrin and bleomycin, respectively. H2O2 induced a lower frequency of mutants, 20-30 per 10(5), for enzymatically generated or bolus, respectively. For the following treatments, mutant frequency at 50% survival is shown. Incubation with human granulocytes induced a low frequency of mutants (about 15 per 10(5)). RNS was tested using a series of NO-donating drugs. Spermine/NO. induced cytotoxicity but no mutants while S-nitroso-N acetylpenicillamine induced a low level, 10 per 10(5). Both release nitrogen monoxide spontaneously, with a t1/2 < 3 h. Glyceryl trinitrate and sodium nitroprusside are two drugs that were slowly metabolized by MN-11 cells (> 12 h). Glyceryl trinitrate induced about 20 per 10(5) while nitroprusside induced 50 per 10(5). Our results indicate that RNS can readily induce mutations detectable in MN-11 cells. At equicytotoxic doses, the induced mutant frequency varied considerably for different drugs, suggesting that different states of nitrogen monoxide (such as NO+ or NO.) may be generated and these may vary in their mutagenic/cytotoxic potential. PMID- 9357554 TI - Elevated levels of the Kearns-Sayre syndrome mitochondrial DNA deletion in temporal cortex of Alzheimer's patients. AB - A mitochondrial hypothesis of Alzheimer's disease (AD) has been proposed based on a number of studies which establish altered oxidative phosphorylation (OXPHOS) and ATP synthesis in AD tissue. Four out of five complexes in the OXPHOS pathway are partly encoded by mitochondrial DNA (mtDNA); thus, this may be a crucial site of lesions that alter brain activity. We examined temporal cortex autopsy tissue for deleted mtDNA by PCR-based methods and Southern analysis. AD tissue was obtained from autopsy-confirmed cases that had a postmortem delay ranging from 5 to 27 h. Using a rat brain model system to examine postmortem effects by Southern analysis, no evidence of mtDNA degradation after 30 h of postmortem delay at room temperature was found. Nine tissue samples taken from AD autopsy brain (average age 68 years) and nine age-matched controls (average age 66 years) were assessed by serial dilution PCR for the 5 kb deletion (mtDNA delta 4977) previously associated with Kearns-Sayre syndrome. Using this method we determined that AD temporal cortex had a 6.5-fold greater frequency of mtDNA delta 4977 than controls (0.0593% vs. 0.0092%, p = 0.0269, one-tailed; p = 0.0530, two-tailed), indicating that damaged mtDNA preferentially accumulates in AD compared to aged brain. PMID- 9357555 TI - Comparison of DNA damage in peripheral blood and spleen lymphocytes using the single-cell gel assay. AB - The alkaline single-cell gel (SCG) or 'comet' assay has been applied to the detection of DNA damage from a number of chemical and biological factors in vivo and in vitro. In the past, a number of cell types has been used with peripheral blood lymphocytes being the most readily accessible. This study was designed to determine whether lymphocytes sequestered in the spleen might prove more sensitive to DNA damage than those in the peripheral circulation. This would result in a more effective SCG assay. Baseline DNA length to width ratios for peripheral blood and splenic lymphocytes did not differ significantly from each other (1.27 and 1.21, respectively). Neither did ratios of lymphocytes from the two sources, sampled 20 and 48 h after injection with 100 mg/kg methyl methanesulfonate (MMS) (3.81 and 3.62 at 20 h, respectively; and 1.96 and 2.21 at 48 h, respectively). Recovery from MMS damage at 168 h postinjection was also not different in the two groups of cells (1.13 and 1.16, respectively). However, an examination of cell profiles of DNA damage showed that splenic lymphocytes had a significantly higher percentage of damaged cells (63.33%) than did peripheral blood lymphocytes (40.67%) 48 h postinjection. Of the hypotheses proposed for this difference, the most likely seems to involve the different proportions of B- and T-lymphocytes present in the peripheral blood and the spleen. Since the difference between peripheral blood and splenic lymphocytes was seen only at 48 h postinjection, the use of splenic lymphocytes in the SCG assay is not advantageous under most circumstances. PMID- 9357556 TI - Euglena gracilis as an eukaryotic test organism for detecting mutagens and antimutagens. AB - The unicellular flagellate Euglena gracilis was used in order to assess the inhibition of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and N-methyl-N nitrosourea (MNU) mutagenicities, which induce white mutants due to the irreversible loss of chloroplasts. All tested compounds, including o-aminobenzoic acid and p-aminobenzoic acid, salicylic acid, acetylsalicylic acid, sodium salicylate and p-aminosalicylic acid, were not mutagenic per se and inhibited MNNG mutagenicity by at least 50%. The last two compounds inhibited by at least 50% also MNU mutagenicity. PMID- 9357557 TI - Evaluation of the genotoxic potential of alkylalkanolamines. AB - Three alkylalkanolamines, N,N-dimethylethanolamine, N-methyldiethanolamine, and tert-butyldiethanolamine, were evaluated for potential genotoxic activity using the Salmonella/microsome reverse gene mutation test, the CHO/HGPRT forward gene mutation test, a sister chromatid exchange test in cultured CHO cells, and an in vivo peripheral blood micronucleus test in Swiss-Webster mice. None of the three alkylalkanolamines produced any significant or dose-related increases in the frequencies of mutations, sister chromatid exchanges or micronuclei. These results indicate that N,N-dimethylethanolamine, N-methyldiethanolamine, and tert butyldiethanolamine are not genotoxic in the tests conducted. PMID- 9357558 TI - Heavy metal content and mutagenic activity, evaluated by Vicia faba micronucleus test, of Tiber river sediments. AB - Tiber river sediment samples, collected in October 1995, were tested for mutagenicity by the micronucleus test in Vicia faba root tips. Four stations were studied within the urban area of Rome: (1) Castel Giubileo, at the entry of the urban area; (2) Ponte Tor di Quinto, immediately after the confluence of the tributary river Aniene; (3) Ponte Sublicio, in the middle of the city; and (4) Ponte della Magliana, immediately outside Rome, 20 km from the sea. Since no significant increase in micronucleus frequency was observed in any of the tested stations compared to control (while in previous campaigns mutagenic activity was observed in some of the same stations), it can be assumed an interesting recovery from mutagenic pollution in the 2 last years. The samples were analysed for pH value, nitrogen, organic matter and carbonate content, and the concentration of some potentially mutagenic heavy metal ions (Zn, Cd, Ni, V, Cu) was assessed. In all samples, a concentration of heavy metals higher than unpolluted areas was observed. However, the alkaline pH measured should keep them as non-bioavailable elements. PMID- 9357559 TI - Genotoxicity induced in human lymphoblasts by atmospheric reaction products of naphthalene and phenanthrene. AB - The genotoxic risks from exposure to polycyclic aromatic hydrocarbons (PAH) have long been recognized. Less well understood are the potential genotoxic risks of the atmospheric reaction products of this class of compounds. In this investigation, we have utilized several human cell genotoxicity assays to evaluate naphthalene, phenanthrene, and their atmospheric reaction products 1 nitronaphthalene, 2-nitronaphthalene, 1-hydroxy-2-nitronaphthalene, 2-hydroxy-1 nitronaphthalene, 1,4-naphthoquinone and 2-nitrodibenzopyranone. In addition, reaction products of naphthalene were generated in a 6700-1 Teflon environmental chamber, collected on a solid adsorbent, extracted and fractionated by normal phase HPLC. Individual fractions were then analyzed using GC-MS, and tested for genotoxicity. Genotoxicity was determined using the human B-lymphoblastoid cell line, MCL-5, which expresses several transfected P450 and epoxide hydrolase genes. Mutagenicity was evaluated at both the heterozygous tk locus and the hemizygous hprt locus, permitting detection of both intragenic and chromosomal scale mutational events. Test compounds were also screened using the CREST modified micronucleus assay. Genotoxicity results indicate that 2 nitronaphthalene and 2-nitrodibenzopyranone possess greater mutagenic potency than their parent compounds, and interestingly, both compounds induced significant increases in mutation frequency at tk but not hprt. These results suggest a mechanistic difference in human cell response as compared to bacteria, where both compounds were previously shown to induce point mutations in the Salmonella reversion assay. The genotoxicity of 2-nitronaphthalene and 2 nitrodibenzopyranone in human cells, together with their high concentrations in ambient air relative to nitro-PAH directly emitted from combustion sources, emphasizes the need to consider atmospheric reaction products of PAH in genotoxicity assessments. PMID- 9357560 TI - Development of a mouse model for studying in vivo T-cell receptor mutations. AB - An experimental system was established to study in vivo T-cell receptor alpha beta (TCR) mutations in murine CD4+ T-lymphocytes. The frequency of TCR-defective mutant T-cells that have the CD3-4+ surface phenotype, was measured using two color flow cytometry of splenic T-cells passed through nylon wool. The spontaneous TCR mutant frequency (MF) in BALB/c mice (2.3 x 10(-4)) was significantly lower than the frequencies of C57BL/6 (4.0 x 10(-4)) and C3H/He (4.2 x 10(-4)) mice. The general trend of the TCR MF started to increase at 3 days after whole-body X-irradiation, reached a peak level at 2-3 weeks, and then gradually decreased with a half-life of about 2 weeks. To analyze how the dose responses for each strain of mouse differed 2 weeks after X-irradiation, the TCR MF dose responses were fitted to a linear-quadratic or a quadratic curve. The coefficients of the quadratic terms in both models for BALB/c mice were significantly higher than those for the other two strains. These findings suggest that some genetic factor(s) may control the susceptibility of somatic genes to both spontaneous and radiation-induced mutagenesis. Establishing an animal model for in vivo TCR mutations will contribute to the clarification of certain unresolved aspects of TCR mutagenesis in humans and will further advance knowledge of screening for environmental mutagens. PMID- 9357561 TI - Detection of in vivo genotoxicity of 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H] furanone (MX) by the alkaline single cell gel electrophoresis (Comet) assay in multiple mouse organs. AB - We tested the genotoxicity of 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H] furanone (MX) in the mouse in 6 organs (liver, lung, kidney, brain, spleen, and bone marrow) and in the mucosa of stomach, jejunum, ileum, colon, and bladder using the alkaline single-cell gel electrophoresis (SCG) (Comet) assay modified by us. Mice were sacrificed 1, 3, 6, and 24 h after oral administration of the mutagen at 100 mg/kg. MX yielded statistically significant DNA damage in the liver, kidney, lung, and brain and in all the mucosa samples. While DNA damage persisted in the gastrointestinal and urinary tract for 6-24 h after a single oral dosing, it peaked in the liver at 1 h and returned to almost the control level at 3 h. Our present results suggest that MX is genotoxic for various mouse organs, but not for the hematopoietic system, and that the alkaline SCG assay with a homogenization technique can be used to predict genotoxicity in the gastrointestinal and urinary tracts. PMID- 9357562 TI - Spontaneous and radiation-induced micronuclei in erythrocytes from four species of wild rodents: a comparison with CBA mice. AB - Almost 100 animals of 4 different species of small wild rodents (bank vole, Clethrionomys glareolus; field vole, Microtus agrestis; yellow-necked mouse, Apodemus flavicollis; and wood mouse, Apodemus sylvaticus) were trapped in central Sweden and used in experiments to determine the spontaneous and radiation induced frequencies of polychromatic (fMPCE) and normochromatic erythrocytes (fMNCE) from bone marrow (bm) and peripheral blood (pb) using flow cytometric analysis. The results were compared with those from similar experiments with CBA mice. The saving of time and labour by the use of the flow cytometer-based analysis was a prerequisite for this study in which about 135 million PCE were analysed. The two species of voles had a mean background fMPCE (bm) of about the same value as CBA mice, while the yellow-necked mice had about five times higher fMPCE (bm). Wood mice had more than twice the fMPCE (bm) compared to CBA mice. Between individual animals in each of the 4 species, the background fMPCE (bm) varied more than between individual CBA mice, and the elimination of micronucleated erythrocytes was considerable. When exposed to ionizing radiation, the voles did not show a significant response. The response of the two Apodemus species was similar to that of the CBA mice, although it varied between individual animals and was not correlated to their background fMPCE. This study indicates that bank voles and field voles are unsuitable testing objects in the in vivo micronucleus assay. On the other hand, yellow-necked mice and wood mice seem to be useful in this test. Since the variation between individuals is considerable in wild Apodemus mice, large groups will be needed for obtaining statistically significant results when exposure to a genotoxic agent is low. Alternatively, repeated samples can be taken from individual wild mice to study the effect of a decreased exposure after keeping the animals for a period of time in an uncontaminated environment. PMID- 9357563 TI - Quantification of mutagenic/carcinogenic heterocyclic amines, MeIQx, Trp-P-1, Trp P-2 and PhIP, contributing highly to genotoxicity of river water. AB - Four mutagenic/carcinogenic heterocyclic amines (HCAs), 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline (MeIQx), 3-amino-1,4-dimethyl-5H-pyrido[4,3 b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido[4,3-b]indole(Trp-P-2) and 2-amino 1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), in organic extracts obtained by blue rayon hanging method from the Yodo River water were quantified. Blue rayon extracts obtained were separated in two stages of fractionation by reversed-phase high performance liquid chromatography (HPLC), and the quantification of corresponding fractions was performed by HPLC with an electrochemical detector for MeIQx and a fluorometric detector for Trp-P-1, Trp-P-2 and PhIP. The geometrical mean values of MeIQx, Trp-P-1, Trp-P-2 and PhIP in extracts collected at 11 locations from the Yodo River systems were 4.8, 26.9, 37.3, and 11.9 ng/g blue rayon equivalent, respectively. The total amounts of four HCAs accounted for mean 24% of the genotoxicity of blue rayon extracts evaluated by the umu test using an O-acetyltransferase-overproducing strain NM2009. PMID- 9357564 TI - Improvement of carcinogen detection in the BALB/3T3 cell transformation assay by using a rich basal medium supplemented with low concentration of serum and some growth factors. AB - To improve the detection sensitivity and reproducibility of the transformation assay using BALB/3T3 cells, we scrutinized a new assay method in which the cells were replated in a medium containing a low concentration of serum after carcinogen treatment. Dulbecco's modified Eagle's medium plus Ham F12 (DME.F12) supplemented with a mixture of insulin, transferrin, ethanolamine and sodium selenite (ITES) and a low concentration of fetal calf serum (FCS) caused transformation at a high frequency with a high reproducibility. The transformation frequency in the culture treated with N-methyl-N'-nitro-N nitrosoguanidine was the highest in DME.F12 medium containing ITES and 2% FCS, and it decreased at either a lower or higher FCS concentration. Moreover, the transformation frequency was not markedly influenced by the difference in lot of FCS, probably due to reduction in FCS concentration. According to the present method, the transformation frequency was 2-times higher and transformed foci appeared much earlier than by the method using the original medium. Next, we examined some geno- and non-genotoxic carcinogens, and some genotoxic non carcinogens to confirm the availability of this method for predicting potential carcinogens. This method could precisely identify not only genotoxic carcinogens but also non-genotoxic carcinogens and genotoxic non-carcinogens. In conclusion, these findings suggest that this method is useful for detection of chemical carcinogens because it provides high sensitivity, high reproducibility and accurate predictivity, without the requirement of 12-O-tetradecanoylphorbol 13 acetate as a promoter, making it harmless to examiner. PMID- 9357565 TI - Hypothermia induces micronuclei in mouse bone marrow cells. AB - We investigated the effect of hypothermia on micronucleus induction in mouse bone marrow cells. Reserpine, which was negative in an in vitro chromosome aberration test, was administered intraperitoneally at 1, 5, 10, 100, and 1000 mg/kg to mice to induce hypothermia. Doses of 10-1000 mg/kg decreased rectal temperature to less than 33.3 degrees C from 24 h to 96 h after dosing and produced a statistically significant (p < 0.01) increase in micronucleated polychromatic erythrocyte frequencies (4.0-12.0/1000). When mice that were administered reserpine at 50, 100, or 200 mg/kg were exposed to an environmental temperature of 30 degrees C for 40 h to keep their body temperature within normal range, the frequency of micronucleated erythrocytes did not increase, while it did without increased environmental temperature. In addition, relatively large micronuclei (diameter of micronucleus > or = 1/4 diameter of cytoplasm) accounted for approximately 50% of the induced micronuclei. The results suggest that the low body temperature of less than 33 degrees C for 40 h induced micronuclei in bone marrow cells, and one possible mechanism was disturbance of the mitotic apparatus. PMID- 9357566 TI - Detection of DNA lesions induced by chemical mutagens by the single cell gel electrophoresis (Comet) assay. 1. Relationship between the onset of DNA damage and the characteristics of mutagens. AB - We evaluated the relationship between the onset of DNA damage and the characteristics of 5 model chemical mutagens with the single-cell gel electrophoresis (SCG) assay using L5178Y mouse lymphoma cells. We treated the cells with each chemical for 3 h and sampled them 0, 21, and 45 h after treatment. DNA damage induced by UV mimetic mutagens MMS and MNU, and X-ray mimetic mutagen BLM was observed just after treatment, crosslinking agent MMC induced DNA damage was detected 21 h after treatment, and 6-MP as an inhibitor of DNA synthesis did not induce DNA damage at any sampling time. These results suggest that the SCG assay detects DNA lesions just after treatment with UV and X ray mimetic mutagens, but needs a waiting period after treatment with crosslinking agents. PMID- 9357567 TI - Detection of DNA lesions induced by chemical mutagens using the single-cell gel electrophoresis (comet) assay. 2. Relationship between DNA migration and alkaline condition. AB - The alkaline condition is an important factor for the alkaline single-cell gel electrophoresis (SCG) assay to detect the genotoxic effects of chemicals. In order to understand the relationship between DNA migration and alkaline condition, the effect of 13 model chemical mutagens with different modes of action was evaluated with the alkaline SCG assay under two different alkaline conditions (pH 12.1 and 12.6). CHO cells were sampled just after treatment for 1 h. The X-ray mimetic mutagen BLM increased DNA migration at pH 12.1 and 12.6 and the results were the same at both pH values. Six alkylating mutagens MNU, ENU, MNNG, ENNG, MMS, and EMS and one base adduct inducer 4-NQO induced a dose dependent response only at pH 12.6. Two DNA crosslinking agents, MMC and DDP, and AMD had negative results. MMC and DDP, however, reduced the positive response of BLM, suggesting that DNA crosslinks could be detected. These results demonstrated that the alkaline condition was important factor for the alkaline SCG assay to detect the genotoxic effects of chemicals. PMID- 9357568 TI - Possible mechanisms of antimutagens by various teas as judged by their effects on mutagenesis by 2-amino-3-methylimidazo[4,5-f]quinoline and benzo[a]pyrene. AB - Possible mechanisms of antimutagenicity of various tea extracts (green, pouchong, oolong and black tea) toward 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and benzo[a]pyrene (B[a]P) were investigated using a Salmonella/microsome assay. Tea extracts exhibited no inhibitory effects toward IQ and B[a]P in bio-antimutagenic assays, indicating that their antimutagenic activity is desmutagenic in nature. The mutagenicities of IQ and B[a]P decreased as the reaction periods of tea extracts with promutagens, S9 mix, or mutagen metabolites increased. The antimutagenicity of tea extracts toward IQ could be attributed (primarily) to an interaction between tea extracts and S9 mix. Apart from their interaction with S9 mix, tea extracts also exhibited antimutagenicity by markedly decreasing the mutagenicity of B[a]P metabolites. These results suggest that tea extracts: (1) inhibit the cytochrome P-450-mediated metabolism of IQ and B[a]P to their ultimate mutagenic metabolite form; and (2) interact with both promutagens and their metabolites in such a way as to reduce their mutagenic potentials. Therefore, the antimutagenic actions of tea extracts are due to a combination of the above distinctive mechanisms, and can vary with the type of mutagen under test. PMID- 9357569 TI - Comparative mutagenic and genotoxic effects of three propionic acid derivatives ibuprofen, ketoprofen and naproxen. AB - The mutagenicity of three propionic acid derivatives, namely ibuprofen, ketoprofen and naproxen, was tested in the Ames mutagenicity assay (in strains TA97a, TA100 and TA102) and in vivo genotoxicity was tested by sister chromatid exchange (SCE) in bone marrow cells of mice. These are the anti-inflammatory drugs frequently used in different parts of the world. Mutagenicity results showed no mutagenic effects in strains TA97a, TA100 and TA102 for all three drugs. Results of in vivo SCE assays indicate that these three drugs are weakly genoxic in bone marrow cells of mice. This is the first report of the Ames mutagenicity assay for ketoprofen and in vivo SCE assay for three drugs. PMID- 9357570 TI - Detection of genotoxicity of polluted sea water using shellfish and the alkaline single-cell gel electrophoresis (SCE) assay: a preliminary study. AB - We exposed two species of shellfish, Patunopecten yessoensis and Tapes japonica, for 4 h to artificial sea water in which N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), ethyl nitrosourea (EMS), 3-chloro-4-dichloromethyl-5-hydroxy-2(H) furanone (MX), or benzo[a]pyrene (B[a]P) were dissolved. We then assessed the DNA damage in cells isolated from the gills using the alkaline single-cell gel electrophoresis (SCG) assay. A statistically significant increase in DNA damage was observed for all exposures. Therefore, the alkaline SCG assay detected DNA damage in gill cells produced by direct mutagens and promutagen dissolved in sea water. T. japonica was exposed to sea water sampled from two Pacific Ocean coasts of Japanese local cities--Hachinohe (Aomori Prefecture, Tohoku) and Nakatsu (Oita Prefecture, Kyushu)--and three bay coasts of the industrial megalopolises--Tokyo, Osaka, and Kobe. A significant increase in DNA damage was observed after the exposure to sea water from Tokyo, Osaka, and Kobe, but not from Hachinohe and Nakatsu. These results suggested the utility of the alkaline SCG assay with shellfish gill cells for monitoring sea water genotoxicity. PMID- 9357571 TI - The micronucleus assay in the zebra mussel, Dreissena polymorpha, to in situ monitor genotoxicity in freshwater environments. AB - Caged zebra mussels, Dreissena polymorpha, were transplanted to 6 monitoring sites receiving industrial effluents suspected of containing genotoxic chemicals. After a residence time of 2 months, the induction of micronuclei (MN) in haemocytes was determined as a criterion for genetic damage. The mean MN frequencies observed in mussels exposed to effluents ranged between 5.0 and 8.8/1000. These rates were significantly higher than the baseline level of 2.0/1000 recorded in a concurrent control mussel group reintroduced at the reference location. Biological fitness descriptors (mortality, attachment biotest, condition index, gonadic index) revealed no relationship between the general well-being of the mussels exposed under contrasted environmental conditions and the frequency of MN induced in their haemocytes. The biological feasibility of the transfer technique of zebra mussels, together with a moderate, but significant, inducibility of MN, are major features towards the development of a first tool for in situ monitoring of genotoxicity in freshwater environments using a common indicator species. PMID- 9357572 TI - Cytogenetic effects of 935.2-MHz (GSM) microwaves alone and in combination with mitomycin C. AB - This paper focuses on the genetic effects of microwaves from mobile communication frequencies (935.2 MHz) alone and in combination with a chemical DNA-damaging agent (mitomycin C). Three cytogenetic endpoints were investigated after in vitro exposure of human whole blood cells. These endpoints were the 'classical' chromosome aberration test, the sister chromatid exchange test and the alkaline comet assay. No direct cytogenetic effect was found. The combined exposure of the cells to the radiofrequency fields followed by their cultivation in the presence of mitomycin C revealed a very weak effect when compared to cells exposed to mitomycin C alone. PMID- 9357573 TI - The herbal preparation abana protects against radiation-induced micronuclei in mouse bone marrow. AB - The induction of micronuclei was studied in mouse bone marrow treated or not with 5, 10 and 20 mg/kg b.wt. of abana (a herbal preparation) before exposure to 0-3 Gy of gamma-radiation. Whole-body irradiation of mice resulted in a dose dependent increase in the frequency of micronuclei. Treatment of mice with various doses of abana before exposure to different doses of gamma-rays resulted in a significant reduction of the micronucleus frequency at all exposure doses. The highest decline in the frequencies of micronuclei was observed after administration of 20 mg/kg abana, where the frequency of micronuclei was approximately 4-fold less than that of the concurrent control. The PCE/NCE ratio was significantly higher in the drug-treated group compared to DDW + irradiated control and it was almost restored to normal level after administration of 20 mg/kg abana. Our results demonstrate that abana protects mice against radiation induced micronucleus formation and radiation-induced decline in cell proliferation. PMID- 9357575 TI - A new method for the enrichment of single renal proximal tubular cells and their first use in the comet assay. AB - A protocol was developed to isolate and enrich single renal proximal tubular cells, performing the following steps: in situ kidney perfusion; isolation of renal tissue pieces by collagenase digestion; selective enrichment of proximal tubular fragments by Percoll gradient centrifugation; and isolation of single proximal tubular cells by digestion of proximal tubular fragments with trypsin. The mean enrichment rate, determined by the glucose-6-phosphatase staining method, was 78.9% with a mean cell viability of 93.8%. After modification of the comet assay protocol, genotoxicity in proximal tubular cells could be investigated. A dose-dependent genotoxic effect of ethyl methanesulphonate in these cells was proven. PMID- 9357576 TI - The resurgence of rubella? PMID- 9357574 TI - Characterisation of genotoxic properties of 2',2'-difluorodeoxycytidine. AB - The genotoxic properties of 2',2'-difluorodeoxycytidine (dFdC) were characterised using diploid, mortal low-passage fibroblasts (LPF cells) and the spontaneously transformed fibroblast cell line V79. In both cell types, incorporation of dFdC into the DNA led to an increase of DNA single-strand breaks evaluated by an in situ nick translation assay and to an accumulation of cells in the S-phase of the cell cycle. At concentrations below those leading to cell cycle arrest, dFdC neither induced sister chromatid exchange (SCE) nor structural chromosome aberrations in LPF cells, whereas V79 cells accumulated SCEs as well as chromosome breaks over a broad dose range. In LPF cells treated with dFdC, chromosomal alterations were detected by the micronucleus assay within a narrow concentration range, whereas in V79 cells, a dose-dependent increase in the appearance of micronuclei was seen up to cytotoxic concentrations. In addition, V79 cells went into apoptosis, as evaluated by nuclear fragmentation and condensation, whereas this phenomenon was not detectable in LPF cells. PMID- 9357577 TI - Capturing clinical reports in a large academic medical center: feeding a central patient data repository. AB - Clinical reports, notes, and other narratives are highly used components in the patient record. Unfortunately, the methods by which these reports are generated are as diverse as the fiscal autonomy of academic clinical departments in a university-based health science center. In this paper, we report on electronically capturing clinical reports, notes, and other text fragments from several hospital sources and many outpatient clinics. The purpose of the capture is to feed the ACIS (Advanced Clinical Information System) central patient data repository that is in use at the University of Utah Health Sciences Center (UUHSC). A survey conducted in early 1994 indicated that about 917,150 reports were generated per year at UUHSC representing about 1.2 million pieces of paper, occupying about 2.3 gigabytes of storage. The most crucial problem encountered in capturing the reports was linking them to the proper patient. Systems that had functioning and well-maintained admit-discharge-transfer (ADT) information performed well, but systems that relied on the human dictator to identify patients, produced patient linkage errors. In our open loop telephone dictation systems this error rate averaged between 6 and 10%. Subsequent to the wide-spread availability of clinical reports on ACIS, this error rate dropped to 3-5%, presumably due to increased demand for on-line availability of this information. From clinical secretaries who use their word processor to create the clinical reports, the linkage error rate was < 1% due to the use of our Advanced Text Upload (ATU) utility. The clinical text component in ACIS contributed significantly to the success of a JCAHO site visit in December 1995. PMID- 9357578 TI - Use of relational database management system by clinicians to create automated MICU progress note from existent data sources. AB - We designed and built an application called MD Assist that compiles data from several hospital databases to create reports used for daily house officer rounding in the medical intensive care unit (MICU). After rounding, the report becomes the objective portion of the daily "SOAP" MICU progress note. All data used in the automated note was available in digital format residing in an institution wide Sybase data repository which had been built to fulfill data needs of the parent enterprise. From initial design of target output through actual creation and implementation in the MICU, MD Assist was created by physicians with only consultative help from information systems (IS). This project demonstrated a method for rapidly developing time saving, clinically useful applications using a comprehensive clinical data repository. PMID- 9357579 TI - Progress notes model. AB - The largest part of the medical record today consists of notes documenting the care delivered to patients and the clinical events relevant to diagnosis and treatment. These "progress notes" serve as the repository of medical facts and clinical thinking, and are intended as a concise vehicle of communication about a patient's condition to those who access the health record. They should be readable, easily understood, complete, accurate, and concise. They must also be flexible enough to logically convey to others what happened during an encounter, e.g., the chain of events during the visit, as well as guaranteeing full accountability for documented material, e.g., who recorded the information and when it was recorded. This paper describes a model for progress notes, which addresses the above needs, and outlines the rationale and principles which led to that model. PMID- 9357580 TI - The rubber meets the road: integrating the Unified Medical Language System Knowledge Source Server into the computer-based patient record. AB - Ongoing improvements in the content of the Unified Medical Language System, coupled with the recent release of the Internet-based Knowledge Source Server (KSS), have prompted us to develop an interface between the KSS and our computer based patient record. We confronted many challenges while developing a robust interface to an Internet-based server, and in integrating the process of codification into the workflow of clinicians. An initial evaluation of the interface in the clinical environment suggests that acceptable performance is attainable. The benefits of using an Internet-accessible tool in a clinical information system appear to justify the effort required. PMID- 9357581 TI - Generic database design for patient management information. AB - Patient management information tracks general facts about the location of the patient and the providers assigned to care for the patient. The Clinical Data Repository at Columbia Presbyterian Medical Center employs a generic schema to record patient management events. The schema is extremely simple, yet can support several different views of patient information, as required by different applications: a longitudinal view of patient visits, including both inpatient and outpatient encounters; a visit-oriented view, to record facts related to a current encounter; a location-based view to provide a census of a nursing ward; and a provider-based view to give a list of the patients currently being cared for by a given clinician. All of these views can be supported in a highly efficient manner by the use of appropriate indexes. PMID- 9357582 TI - Dynamic link between ECG and clinical data by a CORBA-based query engine and temporal mapping. AB - It is important to create a dynamic link method to link distributed patient data across multiple hospitals on an "as needed" basis because the pre-defined links (an item of data has a character or group of characters that indicates the storage of another item of data) are difficult to be managed, or can only be established in part, or are not necessary to be pre-defined in many cases, especially in linking the descriptive data such as history data with the corresponding examination data across multiple hospitals. A method of linking electrocardiogram (ECG) with clinical data dynamically in a Common Object Request Broker Architecture (CORBA) environment has been achieved and verified in a real computing environment to approach to this goal. By this method, distributed patient data can be linked dynamically by a CORBA-based query engine and temporal mapping no matter where they are located on the Internet. The necessary temporal information is provided by either computing or human being. Since multiple time axes for different databases are involved in, some temporal reasoning methods (such as mapping occurrences across temporal contexts and determining bounds for absolute occurrences, etc.) are applied to this study, and a series of temporal mappings including the first mapping, the secondary mapping, the contextual mapping, the extended mapping, the previous mapping and the next mapping are created. In comparison with the pre-defined link, the major strengths of this method are the dynamic link on an "as needed" basis, no limitation of institutional boundaries, easy creation, simplifying the data storage, and the high flexibility, etc. PMID- 9357583 TI - Development of an enterprise-wide clinical data repository: merging multiple legacy databases. AB - We describe the development of a clinical data repository whose core consists of four years of inpatient administrative and billing data from the mainframe legacy systems of the University of Virginia Health System (UVAHS). To these data we have linked a cardiac surgery clinical database and our physician billing data (inpatient and outpatient). Other databases will be merged in the future. A relational database management system (Sybase) running on a dedicated IBM RS/6000 minicomputer was employed to assemble 2.5 Gigabytes of core data describing approximately 100,000 hospital admissions over the four year period. To enable convenient data queries, the system has been equipped with a custom-built WWW user interface, which generates Structured Query Language (SQL) automatically. We illustrate the rapid reporting capabilities of the resulting system with reference to patients undergoing coronary artery bypass graft surgery (CABG). We conclude that this information system: a) constitutes a convenient and low-cost method to increase data availability across the UVAHS; b) provides clinicians with a tool for surveillance of patient care and outcomes; c) forms the core of a comprehensive database from which clinical research may proceed; d) provides a flexible interface empowering a wide variety of clinical departments to share and enrich their own clinical data. PMID- 9357585 TI - Data privacy and confidentiality in the public arena. AB - Public policy debates concerning the collection of healthcare information for use as aggregate databases to underpin healthcare planning are growing increasingly rankerous. Provider concerns for future patient relationships and public fear of damages resulting from information disclosure are driving the development of data collection policy. Through a special Task Force, the State of Maryland has addressed these issues and developed policy recommendations specific to data collection and use. PMID- 9357584 TI - CytoBase: an electronic medical record for cervical cytology. AB - A partnership of medical laboratories in Ontario, Canada has developed and made operational a centralized interactive database of cervical cytology (Pap) reports. This system automatically registers some 60,000 reports monthly, which are submitted electronically from geographically diverse sources. Patient histories are provided on-line to authorized pathologists to support ongoing diagnoses. A formalized coding system has been developed to represent elements of Pap reports in machine readable format, including diagnosis, clinical information about the patient, specimen adequacy, methods of specimen collection and follow up recommendations. The system operates over a private network based on Internet standards. A review of operations, which commenced June 1, 1996, provides insights into applied EMR technology. PMID- 9357586 TI - Design of access control methods for protecting the confidentiality of patient information in networked systems. AB - Public awareness of the potential for violation of personal privacy in clinical information systems is increasing. Much of this increase can be attributed to the popularity and publicity of the World Wide Web. Nightly news reports of intruder break-ins and flaws in Internet software security have stimulated public interest in the security of clinical information systems available over the web. As part of the development of systems designed to provide clinical narratives to physicians over the Internet, we are exploring designs that provide additional protection and security to these systems. Specifically, we are developing and testing automated access control measures based on provider-patient relationships for controlling access to personally identifiable patient information. PMID- 9357587 TI - Guaranteeing anonymity when sharing medical data, the Datafly System. AB - We present a computer program named Datafly that maintains anonymity in medical data by automatically generalizing, substituting, and removing information as appropriate without losing many of the details found within the data. Decisions are made at the field and record level at the time of database access, so the approach can be used on the fly in role-based security within an institution, and in batch mode for exporting data from an institution. Often organizations release and receive medical data with all explicit identifiers, such as name, address and phone number, removed in the incorrect belief that patient confidentiality is maintained because the resulting data look anonymous; however, we show the remaining data can often be used to re-identify individuals by linking or matching the data to other databases or by looking at unique characteristics found in the fields and records of the database itself. When these less apparent aspects are taken into account, each released record can be made to ambiguously map to many possible people, providing a level of anonymity determined by the user. PMID- 9357588 TI - A clinical data repository enhances hospital infection control. AB - We describe the benefits of a relational database of hospital clinical data (Clinical Data Repository; CDR) for an infection control program. The CDR consists of > 40 Sybase tables, and is directly accessible for ad hoc queries by members of the infection control unit who have been granted privileges for access by the Information Systems Department. The data elements and functional requirements most useful for surveillance of nosocomial infections, antibiotic use, and resistant organisms are characterized. Specific applications of the CDR are presented, including the use of automated definitions of nosocomial infection, graphical monitoring of resistant organisms with quality control limits, and prospective detection of inappropriate antibiotic use. Hospital surveillance and quality improvement activities are significantly benefited by the availability of a querable set of tables containing diverse clinical data. PMID- 9357589 TI - Knowledge based functions for routine use at a German university hospital setting: the issue of fine tuning. AB - In this paper we present the introduction of knowledge based functions into clinical routine at Giessen University Hospital. For this purpose a therapy planning module at the medical intensive care unit has been extensively redesigned in order to support the structured documentation of drug prescriptions. After introduction of this new HIS component in January 1996 research has been initiated to establish a basic drug therapy knowledge base. The main components of a knowledge based system have been fully incorporated into the hospital information system WING and are in routine use since December 1996. During a pre-production phase warnings of reminder functions were logged and reviewed by an interdisciplinary team in order to adapt the system to the actual clinical environment. The paper describes experiences during this fine tuning and adaptation process which was necessary to bring a small set of knowledge modules into clinical routine. PMID- 9357591 TI - Using software agents to maintain autonomous patient registries for clinical research. AB - A software agent is an application that can function in an autonomous and intelligent fashion. We have used mobile software agents to maintain clinicians' patient research databases (patient registries). Agents were used to acquire data from the clinician and place it into the registries, copy data from hospital databases into the registries, and report data from the registries. The agents were programmed with the intelligence to navigate through complex network security, interact with legacy systems, and protect themselves from various forms of failure at multiple levels. To maximize the separation between our system and the hospital information infrastructure we often used Java, a platform independent language, to program and distribute our software agents. By using mobile agents, we were able to distribute the computing time required by these applications to underutilized host machines upon which the registries could be maintained. PMID- 9357592 TI - Technique for efficient information retrieval in outpatient systems. AB - In the era of managed care, quality of medical care standards continue to materialize. Most of these standards have long, cumbersome, and complex rules. In light of such problematic rules, efficient ways of retrieving information for a computerized score card are needed. A technique for making such rules less difficult to use is to create Boolean expressions for each quality of care indicator. These Boolean expressions partition the indicators into key words and phrases so that information can be retrieved readily from a system. This study incorporates an outpatient clinical information system of a major university hospital. The technique used to retrieve information and related issues are discussed in the following text. PMID- 9357590 TI - Automatic stenosis detection and quantification in renal arteriography. AB - Visual assessment of the degree of renal artery stenosis on renal arteriography has a large inter- and intraobserver variability. This degree is usually estimated by the ratio between the most narrowed portion of the artery and the reference diameter. The latter is a priori unknown information and thus operator dependent. The objective of the present work was to test the performances of a computer system that was designed to analyze and quantify lesions on 2D renal arteriograms. The main hypothesis was to consider that the most frequent diameter computed along the artery was a good candidate to approximate the reference diameter. Forty nine patient images were collected from the EMMA randomized trial, a multicenter study comparing two treatment strategies in unilateral atheromatous renal artery stenosis of at least 60%. For each image, the degree of stenosis was evaluated by five independent experts and the mean value was used to represent the gold standard for the computer system. The system is based on a fuzzy automaton and performs a syntactic analysis of the arterial segment providing automatic and reproducible quantification of lesions. Both the radiologist caring for the patient and the system were compared to the gold standard. Compared to individual radiologists, the computer system gave a more precise estimation of percent stenosis and did not over or under estimate the severity of the lesion. PMID- 9357593 TI - Automated direct-from-patient information collection for evidence-based diabetes care. AB - BACKGROUND: Computer-based clinical decision support tools can improve physician performance in ambulatory care settings. Acquiring and entering the patient specific information necessary to take advantage of this technology, however, can often be a major impediment. OBJECTIVE: To develop and evaluate a self administered electronic questionnaire for acquiring patient-specific information to be used for generating diabetes-related evidence-based care recommendations. METHODS: An initial paper questionnaire was developed that evaluated current diabetes management, complications, and screening interventions. This was then coded for electronic presentation using software that can also analyze patient responses and produce personal feedback. To evaluate the electronic questionnaire, 47 patients completed it using a small laptop computer and also responded to a personal interview that assessed similar topics. RESULTS: Patients required between 7-29 minutes to complete the questionnaire (mean: 15 min.). For 21 of the 23 topic assessed, the agreement between the electronic questionnaire and the personal interview was 80% or higher. CONCLUSIONS: An electronic, self administered questionnaire can be used to acquire information for generating patient-specific care recommendations. PMID- 9357594 TI - An empirical study of the Health Status Questionnaire System for use in patient computer interaction. AB - Patient involvement in the health care process is very important to any attempt to improve health care quality and patient satisfaction. Although many computerized medical record systems have been introduced, physicians are the only players in the process of data collection and interpretation. A computerized version of the Health Status Questionnaire has been developed to provide a simple, inexpensive method of direct patient entry into the medical record. The system philosophy emphasizes user-centered design and an empirical study was conducted with one hundred twelve outpatients to evaluate the interface aspects of the system as well as the hardware preference of the patients. Statistical analysis indicate that the patients involved in the study rated the user interface of the Health Status Questionnaire System highly. The study also revealed that a considerable number of the general population still have negative preconceptions about their ability to handle a computer or similar looking machinery. When they were asked to use a desktop computer with a mouse, 26 out of 50 patients refused, while 61 out of 62 agreed to use a hand-held pen computer. PMID- 9357595 TI - Clinical care management and workflow by episodes. AB - This paper describes the implementation of clinically defined episodes of care and the introduction of an episode-based summary list of patient problems across Mayo Clinic Rochester in 1996 and 1997. Although Mayo's traditional paper-based system has always relied on a type of 'episode of care' (called the "registration") for patient and history management, a new, more clinically relevant definition of episode of care was put into practice in November 1996. This was done to improve care management and operational processes and to provide a basic construct for the electronic medical record. Also since November 1996, a computer-generated summary list of patient problems, the "Master Sheet Summary Report," organized by episode, has been placed in all patient histories. In the third quarter of 1997, the ability to view the episode-based problem summary online was made available to the 3000+ EMR-capable workstations deployed across the Mayo Rochester campus. In addition, the clinically oriented problem summarization process produces an improved basic "package" of clinical information expected to lead to improved analytic decision support, outcomes analysis and epidemiological research. PMID- 9357596 TI - Using Logical Observation Identifier Names and Codes (LOINC) to exchange laboratory data among three academic hospitals. AB - Using a standard set of names and codes to exchange electronic laboratory data would facilitate multiinstitutional research and data pooling. This need has led to the development of the Logical Observation Identifier Names and Codes (LOINC) database and its test naming convention. We conducted a study which required 3 academic hospitals (in 2 separate medical centers) to extract raw laboratory data from their local information system for a defined patient population, translate tests into LOINC, and provide aggregate data which could then be used to compare laboratory utilization. We found that the coding of local tests into LOINC can often be complex, especially the "Kind of Property" field, and apparently trivial differences in choices made by individual institutions can result in nonmatches in electronically pooled data. In our study, 72-86% of the failures of LOINC to match the same tests between different institutions were due to differences in local coding choices. LOINC has tremendous potential to eliminate the needing for detailed human inspection during the pooling of laboratory data from diverse sites, and perhaps even a built-in capability to adjust matching stringency by selecting subsets of LOINC fields required to match. However, a quality, standard coding procedure at all sites is critical. PMID- 9357597 TI - Medical data mining: knowledge discovery in a clinical data warehouse. AB - Clinical databases have accumulated large quantities of information about patients and their medical conditions. Relationships and patterns within this data could provide new medical knowledge. Unfortunately, few methodologies have been developed and applied to discover this hidden knowledge. In this study, the techniques of data mining (also known as Knowledge Discovery in Databases) were used to search for relationships in a large clinical database. Specifically, data accumulated on 3,902 obstetrical patients were evaluated for factors potentially contributing to preterm birth using exploratory factor analysis. Three factors were identified by the investigators for further exploration. This paper describes the processes involved in mining a clinical database including data warehousing, data query and cleaning, and data analysis. PMID- 9357598 TI - A foundational model of time for heterogeneous clinical databases. AB - Differences among the database representations of clinical data are a major barrier to the integration of databases and to the sharing of decision-support applications across databases. Prior research on resolving data heterogeneity has not addressed specifically the types of mismatches found in various timestamping approaches for clinical data. Such temporal mismatches, which include time-unit differences among timestamps, must be overcome before many applications can use these data to reason about diagnosis, therapy, or prognosis. In this paper, we present an analysis of the types of temporal mismatches that exist in databases. To formalize these various approaches to timestamping, we provide a foundational model of time. This model gives us the semantics necessary to encode the temporal dimensions of clinical data in legacy databases and to transform such heterogeneous data into a uniform temporal representation suitable for decision support. We have implemented this foundational model as an extension to our Chronus system, which provides clinical decision-support applications the ability to match temporal patterns in clinical databases. We discuss the uniqueness of our approach in comparison with other research on representing and querying clinical data with varying timestamp representations. PMID- 9357599 TI - Demystifying mental health information needs through integrated definition (IDEF) activity and data modeling. AB - Today, the process of mental health assessments and treatment are difficult to describe and assimilate into other medical areas. Integrated patient summaries and treatment descriptions are poorly standardized. Any aggregate data analysis must rely on the very few standardized patient data points that may include some demographic information, diagnosis and codable procedures. This paper describes one of the first published attempts to use business process reengineering (BPR) Integrated Computer Assisted Manufacturing Definition (IDEF) activity and data modeling within a focused clinical setting. The clinician's desire to focus on patient care has been used to create both a current and an idealized activity and data model. Clinical patient information was used to build an analyzable database. This provides the potential to track a patient in data throughout a continuum of care. Conceptually, outcomes management is able to use clinical, rather than administrative or claims, data. These models were used to create a prototype for a computer-based patient record which would allow outcomes management. It was tested successfully as a proof of concept. PMID- 9357600 TI - Evaluating large scale health information systems: from practice towards theory. AB - With the introduction of large scale health information systems which are incrementally developed from legacy systems, evaluators are faced with difficult methodological and practical problems. Some of the problems involved in multidisciplinary multi-method evaluations care discussed. It is argued that the development of a framework for evaluation is necessary in order to successfully plan an evaluation, understand the implications of the results and make future predictions based upon them. Some suggestions for arriving at such a framework are put forward. PMID- 9357601 TI - Improving information access with an emergency department system. AB - An emergency department (ED) clinical system was developed by in-house personnel, with ED physician, nursing, registration and clerical staff input. The utilization of existing hardware and customization of the hospital's mainframe hospital information system (HIS) facilitated the implementation of a cost effective system that meets the information access needs of a busy, state-of-the art academic ED. The transition to automation of the ED was facilitated through the use of a comprehensive training plan and change strategies. PMID- 9357602 TI - Design of a general clinical notification system based on the publish-subscribe paradigm. AB - We describe the design and initial implementation of a notification sub-system, as a component of a modern information management architecture. The system, based on the publish-subscribe paradigm, provides a framework of event-based communications for the implementation of various important clinical applications including the notification of alerts and reminders with escalation algorithms, the reliable distribution of documents, and the implementation of intelligent patient-specific monitoring processes. The initial implementation of the system, providing the notification of the unit staff about new orders, indicates that the model is viable both in terms of functionality and ability to scale up. PMID- 9357603 TI - Accuracy and efficiency of an automated system for calculating APACHE II scores in an intensive care unit. AB - We evaluated the reliability and efficiency of an automated system for calculating APACHE II scores. We imported an automated APACHE II scoring system developed at another institution. We scored a convenience sample of 50 consecutive intensive care unit (ICU) admissions using three methods: (1) the automated system (2) an expert scorer using a manual data abstraction method, and (3) the current manual scoring method used in the ICU. We analyzed interrater reliability among the three groups, and compared scoring time between the automated and the expert groups. Interrater reliability testing demonstrated a very high agreement between the computer and the expert scorers, (r = 0.97 p < 0.01) and a high agreement between the computer and the nursing staff (r = 0.80, p < 0.01). The mean time required to complete the data sheet manually and input data into a standalone computer manually was 4 minutes and 55 seconds, compared to total mean time of 33 seconds for the automated system. Automated APACHE II scoring yielded results more reliable than those of an expert scorer, as judged by a second researcher. The time required to score patients was reduced and reliability of scores was improved with use of an automated APACHE II scoring system. PMID- 9357604 TI - Nurses use of health status data to plan for patient care: implications for the development of a computer-based outcomes infrastructure. AB - The purpose of this study was to examine the relationships between the patient's health status at hospital admission and the initial care planned by the nurse. Functional status, engagement in care, and psychosocial well-being were measured by the Health Status Outcome Dimensions(HSOD) instrument. The HSOD is the foundation for developing a computer-based infrastructure for the analysis of health related outcomes. The consecutive, convenience sample of 308 subjects was drawn from five acute clinical populations: pulmonary; cerebrovascular, cardiac; gastrointestinal; and infection. Logistic and multiple regression analyses were used to test the relationships between control (patient and setting) variables, health status, and the dependent variables of type of problem identified, number of problems identified, and the time required to implement interventions ordered for the patient. In seven of ten models, control variables of facility, age, and/or severity of illness contributed to a model at p < .01. In six of ten models, at least one health status measure significantly explained variation beyond the control variables, at p < .01. Study results support using data gathered during the course of care, to evaluate the process of that care. Further work is needed to understand the effects of setting and provider variables on the use of health status data in care planning. Computer-based outcomes infrastructures are essential to support the collection and analysis of health status over time. PMID- 9357605 TI - Documenting 'what nurses do'--moving beyond coding and classification. AB - A variety of strategies for knowledge representation have been applied to the texts from a number of medical domains. Many of the techniques rely on the well defined ways in which medical terms are used within a given domain, a phenomenon referred to as 'sublanguage.' Because much of nursing documentation involves the use of 'everyday' language, the viable application of sublanguage-based approaches to knowledge representation of nursing documentation is not a forgone conclusion. We propose an approach utilizing semantic markup of nursing notes as a strategy for determining whether the documentation of 'what nurses do' is a sublanguage Results of an initial feasibility study utilizing the approach are presented. PMID- 9357607 TI - Cross-institutional reuse of a problem statement knowledge base. AB - This article describes client and server applications for a problem statement knowledge base derived from a large corpus of provider entered terminology. The current status and potential for integration of the server into the Vanderbilt University Medical Center computing environment are discussed. Finally, an experiment in multiple dimensions of reuse for problem list terms is introduced, and possible strategies to mediate between free text and coded data are examined. PMID- 9357606 TI - A patient workflow management system built on guidelines. AB - To provide high quality, shared, and distributed medical care, clinical and organizational issues need to be integrated. This work describes a methodology for developing a Patient Workflow Management System, based on a detailed model of both the medical work process and the organizational structure. We assume that the medical work process is represented through clinical practice guidelines, and that an ontological description of the organization is available. Thus, we developed tools 1) for acquiring the medical knowledge contained into a guideline, 2) to translate the derived formalized guideline into a computational formalism, precisely a Petri Net, 3) to maintain different representation levels. The high level representation guarantees that the Patient Workflow follows the guideline prescriptions, while the low level takes into account the specific organization characteristics and allow allocating resources for managing a specific patient in daily practice. PMID- 9357608 TI - Structuring clinical practice guidelines in a relational database model for decision support on the Internet. AB - The rapid proliferation of clinical practice guidelines (CPGs) has made computerization increasingly useful to clinicians. Computerization, however, requires transformation of the content and logic of each guideline into a computer-accessible form. In this project, we sought to use a relational database to construct a generalized guideline knowledge base for use with Internet-based decision support applications. We hypothesized that knowledge representation schemes could be developed to capture guideline content and logic within the constraints of a relational database model. In this paper we describe a database schema based on a relational model for computerizing CPGs using a hybrid of structured and procedural knowledge representation schemes. We developed and refined this model in the context of five diverse CPGs and found it accommodated all necessary representational requirements. PMID- 9357609 TI - Development and use of a Guideline Entry Wizard to convert text clinical practice guidelines to a relational format. AB - Computerization of clinical practice guidelines (CPGs) has been proposed as one solution to enhance the use of guidelines in influencing standard clinical care. However, the conversion of text guidelines to the format required by a computer program is a major barrier. Clinicians who best understand the content of CPGs are typically ill equipped to convert textual guidelines into a computer accessible format. The potential of knowledge acquisition tools to assist in this process has been documented in the literature. In this paper we describe an application prototype, the Guideline Entry Wizard, created to assist in the conversion of text CPGs to a structured format within a relational database. We have tested this application through the input of information from several CPG. The application is a prototype for a more advanced tool. We have used this prototype to enter several CPGs and have demonstrated its effectiveness in inputting guideline content into a knowledge base. PMID- 9357610 TI - Inducing practice guidelines from a hospital database. AB - Improving health care quality requires the elimination of unnecessary variation in the care process. Decision support applications already exist that can foster adherence to standards. The challenge resides in developing standards consistent with good medical practice. In this paper we present our efforts in determining where sufficient clinical data are captured electronically to automatically define a care process, and what analyses can be done to identify additional data that would allow a care process to be defined. Data routinely collected by a hospital information system have been examined. The analysis tools utilized include logistic regression, a neural network, a Bayesian network, and a rule induction program. PMID- 9357612 TI - The CliniCon framework for context representation in electronic patient records. AB - A well-known problem of current electronic patient records in that they usually fail to represent the semantic relationships between the involved clinical data. This has to be viewed as a problem especially in the domains characterized by a complex and long-term treatment, as the medical decision making process may not be comprehensible anymore from the data entries themselves. Context representation can overcome these limitations, enabling the record to express causality, revisions, conflicts, or individual heuristics explicitly. This article introduces CLINICON which is a formal framework for domain-independent context representation based on Sowa's conceptual graphs. PMID- 9357611 TI - Decision-analytic valuation of clinical information systems: application to an alerting system for coronary angiography. AB - BACKGROUND: Many patients who need coronary angiography fail to get it and they have decreased survival as a result. This study demonstrates the use of decision analysis to predict the survival value of an alerting system for necessary angiography. METHODS: Data on the use of angiography and survival after myocardial infarction (MI) were taken from a published cohort study. The expected value of information (EVI) was calculated for alerts that angiography is necessary. Maximal EVI was estimated by assuming that alert advice is always followed. Sensitivity analysis relaxed that assumption. Hypothetical data were generated to demonstrate EVI analysis for narrower subcohorts. RESULTS: A maximally effective alerting system would increase survival in this cohort by 2.2% over 1-4 years after MI. The system would therefore need to be applied to 46 people to prevent one death. Its effectiveness would decrease linearly with decreasing adherence to its advice. Given sufficiently detailed outcome and prevalence data, EVI analysis could also predict the survival value of the system's individual data elements. CONCLUSIONS: An alerting system that ensures necessary angiography post-MI should have a survival value comparable to the value of t-PA over streptokinase. EVI analysis provides a framework for predicting the overall effectiveness of information systems and for understanding the contribution of individual features to a system's effectiveness. PMID- 9357613 TI - Toward implementation of artificial neural networks that "really work". AB - Artificial neural networks are established analytical methods in bio-medical research. They have repeatedly outperformed traditional tools for pattern recognition and clinical outcome prediction while assuring continued adaptation and learning. However, successful experimental neural networks systems seldom reach a production state. That is, they are not incorporated into clinical information systems. It could be speculated that neural networks simply must undergo a lengthy acceptance process before they become part of the day to day operations of health care systems. However, our experience trying to incorporate experimental neural networks into information systems lead us to believe that there are technical and operational barriers that greatly difficult neural network implementation. A solution for these problems may be the delineation of policies and procedures for neural network implementation and the development a new class of neural network client/server applications that fit the needs of current clinical information systems. PMID- 9357614 TI - Clinical event monitoring at the University of Pittsburgh. AB - Although the literature on event monitoring is extensive, it does not cover all issues that we encountered while developing an event monitor at our institution. We had to resolve issues related to event detection, scalability, what topics were suitable for asynchronous decision support, and overlap of efforts of other groups at the institution attempting to improve quality and lower cost of care. In this paper, we describe our experience deploying CLEM, the clinical event monitor at the University of Pittsburgh with emphasis on these topics. PMID- 9357616 TI - Incremental learning of probabilistic rules from clinical databases based on rough set theory. AB - Several rule induction methods have been introduced in order to discover meaningful knowledge from databases, including medical domain. However, most of the approaches induce rules from all the data in databases and cannot induce incrementally when new samples are derived. In this paper, a new approach to knowledge acquisition, which induce probabilistic rules incrementally by using rough set technique, is introduced and was evaluated on two clinical databases. The results show that this method induces the same rules as those induced by ordinary non-incremental learning methods, which extract rules from all the datasets, but that the former method requires more computational resources than the latter approach. PMID- 9357615 TI - Use of CLIPS for representation and inference in a clinical event monitor. AB - We developed a clinical event monitor that is currently deployed in an inpatient setting. We selected CLIPS as the basis for its KB and inference engine. This paper describes the considerations that went into that decision, how we represented drug and laboratory knowledge in CLIPSs, and how we extended CLIPS to deal with temporal inferences. We also review the published literature about the use of CLIPS in medicine. PMID- 9357617 TI - Modelling cardiac patient set residuals using rough sets. AB - Many medical studies deal with the assessment of the prognostic or diagnostic power of some particular test with respect to some particular medical condition. However, even though a test is deemed to be powerful in this respect, the test may not be strictly needed to perform for everyone. If the test is costly or invasive, this issue is of particular interest. This paper presents a methodology based on rough set theory and Boolean reasoning that can be used to identify those patients for whom performing the test is redundant or superfluous. Furthermore, the methodology enables one to automatically construct a set of descriptive and minimal if-then rules that model the patient group in need of the test. A reanalysis of a previously published real-world dataset of patients with chest pain is used as a case study. PMID- 9357618 TI - An expert-guided decision tree construction strategy: an application in knowledge discovery with medical databases. AB - With the steady growth in electronic patient records and clinical medical informatics systems, the data collected for routine clinical use have been accumulating at a dramatic rate. Inter-disciplinary research provides a new generation of computation tools in knowledge discovery and data management is in great demand. In this study, an expert-guided decision tree construction strategy is proposed to offer an user-oriented knowledge discovery environment. The strategy allows experts, based on their expertise and/or preference, to override inductive decision tree construction process. Moreover, by reviewing decision paths, experts could focus on subsets of data that may be clues to new findings, or simply contaminated cases. PMID- 9357619 TI - Prototype expert system to assist with the stabilization of neonates prior to transport. AB - The transport of sick or premature newborns from community hospitals to acute care facilities is often necessary for the infants to receive the level of care required. Prior stabilization of these infants is imperative to a successful transport. The knowledge required to treat and stabilize sick and premature newborns is specialized and may not be available in community hospitals. "The S.T.A.B.L.E. Assistant" is a prototype rule based decision support system based on the educational program "Transporting Newborns the S.T.A.B.L.E. Way". "The S.T.A.B.L.E. Assistant" accepts clinical information related to an infant's breathing status, blood glucose status and selected lab values and provides instructions as to what interventions are needed to stabilize and prepare the infant for transport. The computerized program was evaluated using data collected from 19 charts of newborns requiring transport from a community hospital to a tertiary care center. Patient data were entered into the computerized program and the subsequent instructions were reviewed by a group of neonatology and neonatal transport experts. Reviewers evaluated "The S.T.A.B.L.E. Assistant" for the safety of the program, the extent to which the computerized program follows the educational program guidelines and the degree to which the instructions generated are within the scope of community caregivers' practice. Results were positive, indicating "The S.T.A.B.L.E. Assistant" prototype is safe, and within the guidelines of neonatal clinical practice. PMID- 9357620 TI - Usability testing in medical informatics: cognitive approaches to evaluation of information systems and user interfaces. AB - This paper describes an approach to the evaluation of health care information technologies based on usability engineering and a methodological framework from the study of medical cognition. The approach involves collection of a rich set of data including video recording of health care workers as they interact with systems, such as computerized patient records and decision support tools. The methodology can be applied in the laboratory setting, typically involving subjects "thinking aloud" as they interact with a system. A similar approach to data collection and analysis can also be extended to study of computer systems in the "live" environment of hospital clinics. Our approach is also influenced from work in the area of cognitive task analysis, which aims to characterize the decision making and reasoning of subjects of varied levels of expertise as they interact with information technology in carrying out representative tasks. The stages involved in conducting cognitively-based usability analyses are detailed and the application of such analysis in the iterative process of system and interface development is discussed. PMID- 9357621 TI - Willingness-to-pay utility assessment: feasibility of use in normative patient decision support systems. AB - The authors developed an automated patient interviewing tool to elicit individuals' willingness-to-pay (WTP) utilities under conditions of uncertainty and examined the reliability of this method and its potential usefulness in clinical decision support. We tested this method in 52 healthy volunteers using a computer-based interview that trained subjects in standard gamble (SG) and WTP methods, and elicited preferences for moderate Gaucher disease using WTP and SG. We assessed the validity of the WTP method by calculating the cost-effectiveness threshold implied by subjects' WTP and SG utilities; we also assessed subjects' understanding and comfort with using WTP for decision making by a questionnaire. The WTP method had good test-retest reliability (r = 0.796), and produced a cost effectiveness ratio and ratings for understanding and clarity that support its validity. Moreover, many subjects felt that WTP was a reasonable (83%) method for therapeutic decision making and expressed comfort (62%) in using the method for their own health care decisions. These results suggest that a probabilistic method for WTP utility assessment is potentially useful for acquiring patient preferences for use in normative decision support systems. PMID- 9357622 TI - Evaluating the potential effectiveness of using computerized information systems to prevent adverse drug events. AB - In this study a dynamic computer simulation model is used to estimate the effectiveness of various information systems applications designed to detect and prevent medication errors that result in adverse drug events (ADEs). The model simulates the four stages of the drug ordering and delivery system: prescribing, transcribing, dispensing and administering drugs. In this study we simulated interventions that have been demonstrated in prior studies to decrease error rates. The results demonstrated that a computerized information system that detected 26% of medication errors and prevented associated ADEs could save 1,226 days of excess hospitalization and $1.4 million in hospital costs annually. Those results suggest that such systems are potentially a cost-effective means of preventing ADEs in hospitals. The results demonstrated the importance of viewing adverse drug events from a systems perspective. Prevention efforts that focus on a single stage of the process had limited impact on the overall error rate. This study suggests that system-wide changes to the drug-ordering and delivery system are required to significantly reduce adverse drug events in a hospital setting. PMID- 9357623 TI - The impact of anticipatory patient data displays on physician decision making: a pilot study. AB - Computerized patient records have long offered the promise of facilitated access to patient data for clinical decision-making. Nonetheless, the decision process benefits of improved patient data access have been poorly quantified by prior informatics research. We conducted a pilot study to test the feasibility of study methods and gather data for the planning of a future clinical trial designed to assess the impact of patient data summary displays on serum lipid test interpretation time, on targeted data retrieval time for related data, and on decision quality. The pilot demonstrated feasibility and high face validity of the decision-making simulation methods used. Problem-focused patient data summaries appear to reduce time-based decision performance measures by 40-50%, and may improve decision quality even without the inclusion of knowledge-based recommendations or guideline representations. PMID- 9357624 TI - New computer-based tools for empiric antibiotic decision support. AB - Since 1995 we have been developing a decision-support model, called Q-ID, which uses a series of infectious disease knowledge bases to make recommendations for empirical treatment or to check the appropriateness of current antibiotic therapy. From disease manifestations and risk factors, a differential diagnosis for the patient is generated by a diagnostic medical expert system. The resulting probability of each: disease is multiplied by the expected benefit in improved mortality and morbidity from optimal antibiotic treatment of each disease. To generate empirical treatment recommendations, site-specific data on sensitivity to antibiotics of each organism is used as an estimate of the likelihood of achieving maximum benefit for each disease on the patient's differential. Combining this data with drug and patient specific factors, the model recommends the antibiotic(s) most likely to produce the optimal benefit in this patient with the least risk and expense. In this paper the model is described, excerpts from each of the knowledge bases are presented, and performance of the model in a real case is shown for illustration. PMID- 9357625 TI - Advanced alerting features: displaying new relevant data and retracting alerts. AB - We added two advanced features to our automated alerting system. The first feature identifies and displays, at the time an alert is reviewed, relevant data filed between the login time of a specimen leading to an alerting result and the time the alert is reviewed. Relevant data is defined as data of the same kind as generated the alert. The other feature retracts alerts when the alerting value is edited and no longer satisfies the alerting criteria. We evaluated the two features for a 14-week period (new relevant data) and a 6-week period (retraction). Of a total of 1104 alerts in the 14-week evaluation, 286 (25.9%) had new relevant data displayed at alert review time. Of the 286, 75.2% were due to additions of comments to the original piece of alerting data; 24.1% were due to new or pending laboratory results of the same type that generated the alert. Two alerts (out of 490) were retracted in a 6 week period. We conclude that in our system, new clinically relevant data is often added between the time of specimen login and the time that an alerting result from that specimen is reviewed. Retractions occur rarely but are important to detect and communicate. PMID- 9357627 TI - Using data mining to characterize DNA mutations by patient clinical features. AB - In most hereditary cancer syndromes, finding a correspondence between various genetic mutations within a gene (genotype) and a patient's clinical cancer history (phenotype) is challenging; to date there are few clinically meaningful correlations between specific DNA intragenic mutations and corresponding cancer types. To define possible genotype and phenotype correlations, we evaluated the application of data mining methodology whereby the clinical cancer histories of gene-mutation-positive patients were used to define valid or "true" patterns for a specific DNA intragenic mutation. The clinical histories of patients with their corresponding detailed attributes without the same oncologic intragenic mutation were labeled incorrect or "false" patterns. The results of data mining technology yielded characterizing rules for the true cases that constituted clinical features which predicted the intragenic mutation. Some of the initial results derived correlations already independently known in the literature, adding to the confidence of using this methodological approach. PMID- 9357626 TI - Evaluation of a decision support system for pressure ulcer prevention and management: preliminary findings. AB - A decision support system for prevention and management of pressure ulcers was developed based on AHCPR guidelines and other sources. The system was implemented for 21 weeks on a 20-bed clinical care unit. Fifteen nurses on that unit volunteered as subjects of the intervention to see whether use of the system would have a positive effect on their knowledge about pressure ulcers and on their decision-making skills related to this topic. A similar care unit was used as a control. In addition, the system was evaluated by experts for its instructional adequacy, and by end users for their satisfaction with the system. Preliminary results show no effect on knowledge about pressure ulcers and no effect on clinical decision making skills. The system was rated positively for instructional adequacy, and positively for user satisfaction. User interviews related to satisfaction supplemented the quantitative findings. A discussion of the issues of conducting experiments like this in today's clinical environment is included. PMID- 9357628 TI - A decision support system for microbiology quality control. AB - Manual review of antibiotic sensitivity testing results is an essential component of a microbiology laboratory's quality control process. Such review is tedious and prone to human error, however. An expert system is described that remembers which susceptibility patterns are considered typical or atypical by expert reviewers, then uses these to prescreen future isolates. It uses a similarity function to allow matching against this library when two patterns are close, but not identical. Use of this system allows more efficient and reliable review of the laboratory's antibiotic sensitivity testing results. PMID- 9357630 TI - A qualitative study of clinicians ways of using a decision-support system. AB - We have studied how clinicians approached a decision-support system to manage patient cases. The design of the system under study was based on an integration of hypertext and rule-based systems. World-Wide Web technology was used for the implementation of the system. By using grounded theory and stimulated recall, we found that getting patient-specific support and continuing medical education were the two major usages of the system and that the three parameters relevance, validity, and work were important in describing how the system was experienced by the users. PMID- 9357629 TI - Changes in diagnostic decision-making after a computerized decision support consultation based on perceptions of need and helpfulness: a preliminary report. AB - We examined the degree to which attending physicians, residents, and medical students' stated desire for a consultation on difficult-to-diagnose patient cases is related to changes in their diagnostic judgments after a computer consultation, and whether, in fact, their perceptions of the usefulness of these consultations are related to these changes. The decision support system (DSS) used in this study was ILIAD (v4.2). Preliminary findings based on 16 subjects' (6 general internists, 4 second-year residents in internal medicine, and 6 fourth year medical students) workup of 136 patient cases indicated no significant main effects for 1) level of experience, 2) whether or not subjects indicated they would seek a diagnostic consultation before using the DSS, or 3) whether or not they found the DSS consultation in fact to be helpful in arriving at a diagnosis (p > .49 in all instances). Nor were there any significant interactions. Findings were similar using subjects or cases as the unit of analysis. It is possible that what may appear to be counter-intuitive, and perhaps irrational, may not necessarily be so. We are currently examining potential explanatory hypotheses in our ongoing current, larger study. PMID- 9357631 TI - Assessing the value of newer pharmacologic agents in non-ST elevation patients: a decision support system application. AB - Newer pharmacologic agents have demonstrated significant clinical and economic benefit in high-risk percutaneous transluminal coronary angioplasty (PTCA) patients. However, the higher costs of these agents may prohibit their use in lower-risk coronary artery disease (CAD) populations. We developed a decision support system (DSS) to determine the level of clinical effectiveness these newer agents must exhibit to be either cost-neutral or cost-effective in non-ST elevation patients. Our DSS evaluated six month cumulative costs, increased years of life saved (YOLS), and lifetime cost-effectiveness. We found that these therapies can cost as much as $1500 and be cost-neutral at six months if they reduce the composite endpoint of death, myocardial infarction (MI), or revascularization by 15%, and they may cost as much as $3000 and be cost effective if they reduce this endpoint by 10%. PMID- 9357632 TI - Mixed integer programming optimization models for brachytherapy treatment planning. AB - Mixed integer programming is proposed as an approach for generating treatment plans for brachytherapy. Two related but distinct, mixed integer programming models are tested on data from eight prostate cancer patients. The results demonstrate that in some cases, "good" treatment plans can be obtained in less than five CPU minutes. PMID- 9357633 TI - Operationalization of clinical practice guidelines using fuzzy logic. AB - There are a number of obstacles to successful operationalization of clinical practice guidelines, including the difficulty in accurately representing a statement's decidability or an action's executability. Both require reasoning with incomplete and imprecise information, and we present one means of processing such information. We begin with a brief overview of fuzzy set theory, in which elements can have partial memberships in multiple sets. With fuzzy inferencing, these sets can be combined to create multiple conclusions, each with varying degrees of truth. We demonstrate a fuzzy model developed from a published clinical practice guideline on the management of first simple febrile seizures. Although the creation of fuzzy sets can be an arbitrary process, we believe that fuzzy inferencing is an effective tool for the expression of guideline recommendations, and that it can be useful for the management of imprecision and uncertainty. PMID- 9357634 TI - HyperCare: a prototype of an active database for compliance with essential hypertension therapy guidelines. AB - HyperCare is a prototype of a decision support system for essential hypertension care management. The medical knowledge implemented in HyperCare derives from the guidelines for the management of mild hypertension of the World Health Organization/International Society of Hypertension, and from the recommendations of the United States Joint National Committee on Detection, Evaluation and Treatment of High Blood Pressure. HyperCare has been implemented using Chimera, an active database language developed at the Politecnico di Milano. HyperCare proves the possibility to use active database systems in developing a medical data-intensive application where inferential elaboration of moderate complexity is required. PMID- 9357636 TI - A temporal database mediator for protocol-based decision support. AB - To meet the data-processing requirements for protocol-based decision support, a clinical data-management system must be capable of creating high-level summaries of time-oriented patient data, and of retrieving those summaries in a temporally meaningful fashion. We previously described a temporal-abstraction module (RESUME) and a temporal-querying module (Chronus) that can be used together to perform these tasks. These modules had to be coordinated by individual applications, however, to resolve the temporal queries of protocol planners. In this paper, we present a new module that integrates the previous two modules and that provides for their coordination automatically. The new module can be used as a standalone system for retrieving both primitive and abstracted time-oriented data, or can be embedded in a larger computational framework for protocol-based reasoning. PMID- 9357635 TI - A prototype Web site for immunization knowledge maintenance. AB - IMM/Web is a prototype Web site designed to assist in the maintenance of the knowledge required to perform computer-based forecasting for childhood immunization. IMM/Web operates in conjunction with IMM/Serve, a immunization forecasting program that takes a child's immunization history and produces recommendations as to which vaccinations are due, and which should be scheduled next. IMM/Serve's domain knowledge is expressed in both tabular and rule-based form. Using IMM/Web, the various tabular forecasting parameters can be modified via the Web. Test cases can then be generated automatically which can be used to help verify the new version of logic. Finally, the test cases can be automatically passed for IMM/Serve to analyze using the newly defined parameters. The IMM/Web project is exploring how the process of updating, customizing, and testing new versions of a computer-based clinical guideline might be performed and guided in an organized fashion via the World-Wide Web. PMID- 9357637 TI - Creating an environment for linking knowledge-based systems to a clinical database: a suite of tools. AB - A difficulty in using knowledge-based systems has been linking them to clinical databases. The challenge is in making a correct mapping from the data in the knowledge base to the data in the database. At Columbia-Presbyterian Medical Center, we have built a suite of tools developed to create queries that address this challenge. The tools were designed to allow users to easily retrieve data from the database without requiring the users have extensive database and vocabulary knowledge. The tools help users write correct queries (Query Builder), find correct terms in the clinical database (MED Browser), aggregate the resulting data into a useful form (Clinical Database Browser), and allow the user to test the query within the environment of the knowledge-based system (Event Playback). The tools have been in use for one year. PMID- 9357638 TI - Towards improved knowledge sharing: assessment of the HL7 Reference Information Model to support medical logic module queries. AB - Because clinical databases vary in structure, access methods and vocabulary used to represent data, the Arden Syntax does not define a standard model for querying databases. Consequently, database queries are encoded in ad hoc ways and enclosed in "curly braces" in Medical Logic Modules (MLMs). However, the nonstandard representation of queries impairs sharing of MLMs, an impediment that has come to be known as the "curly braces problem." As a first step in solving this problem, we evaluated the proposed HL7 Reference Information Model (RIM) as a foundation for a standard query model for the Arden Syntax. Specifically, we analyzed the MLM knowledge base at the Columbia-Presbyterian Medical Center and compared the queries in these MLMs to the RIM. We studied 488 queries in 104 MLMs, identifying 674 total query data elements. Laboratory tests accounted for 45.8% of these elements, while demographic and ADT data accounted for 37.6%. Pharmacy orders accounted for 10.5%, medical problems for 4.3% and MLM output messages for 1.6%. We found that the RIM encompasses all but those data elements signifying MLM output (1.6% of the total). We conclude that the majority of queries in the CPMC knowledge base access a relatively small set of data elements and that the RIM encompasses these elements. We propose extensions of this analysis to continue construction of an Arden query model capable of solving the "curly braces problem." PMID- 9357639 TI - Generation of development environments for the Arden Syntax. AB - Providing appropriate development environments for specialized languages requires a significant development and maintenance effort. Specialized environments are therefore expensive when compared to their general-language counterparts. The Arden Syntax for Medical Logic Modules (MLM) is a standardized language for representing medical knowledge. We have used PROTEGE-II, a knowledge-engineering environment, to generate a number of experimental development environments for the Arden Syntax. MEDAILLE is the resulting MLM editor, which provides a user friendly environment that allows users to create and modify MLM definitions. Although MEDAILLE is a generated editor, it has similar functionality, while reducing the programming effort, as compared to other MLM editors developed using traditional programming techniques. We discuss how developers can use PROTEGE-II to generate development environments for other standardized languages and for general programming languages. PMID- 9357640 TI - Experiences in deployment of a Web-based CIS for referring physicians. AB - Improving the timeliness and efficiency of information exchange between the hospital and clinicians in the health care community is an area of active interest at the Massachusetts General Hospital (MGH). Providing computer-based access to referring physicians who are not formally affiliated with the hospital is a particular challenge since these offices are not connected to the hospital network and lack the standard hospital workstation hardware and software. Installing clients for the hospital's clinical applications at these sites has been a difficult and costly proposition. The emergence of Web technology yields an alternative method for developing clinical applications for this remote, diverse user population. We present our experiences during the first six months of deployment of a Web-based clinical information system designed for use by referring physicians. PMID- 9357641 TI - Do electronic mail discussion lists act as virtual colleagues? AB - Anesthesiology Discussion Group (ADG), an electronic mail (email) discussion list, has previously been shown to be a clinically oriented, cost-effective form of telemedicine. ADG is composed of an international collection of anesthesia providers. Discussions with colleagues are generally informal in nature and are examples of types of information-seeking behavior which frequently occur in hallways or lounges of a hospital or clinic. Information-seeking occurs when a health care provider searches for information which will be used to solve or satisfy a patient's problem or need. We surveyed practitioners who had previously submitted non-rhetorical, clinical questions to the group. After analysis of the questionnaire results, we conclude that ADG is a valuable resource used for information-seeking and is a clinically effective form of telemedicine. Many of the respondents indicated that they used ADG to obtain second opinions from the collective expertise of group members. Respondents also indicated that they were generally satisfied with the quality of responses and would not hesitate to use ADG for future clinical questions. PMID- 9357642 TI - Making effective referrals: a knowledge-management approach. AB - Patients and physicians often choose specially consultants with only limited knowledge of the available options. Access to information about specialists that was directly relevant to patient and clinician preferences could improve the effectiveness of the referral process. We have developed a prescriptive representation of the process of selecting consultants. This "referral map," based on decision theory, uses patient and provider preferences elicited through a literature review and interviews with physicians and provides a formal framework for representing referral knowledge and for evaluating referral options. Our method suggests that the goals and processes of selecting consultants can be managed more systematically using explicit repositories. Such systematic management promises to have a beneficial impact on the delivery of health care, as well as on patient satisfaction. PMID- 9357643 TI - Practical lessons in remote connectivity. AB - Community Health Information Networks (CHINs) require the ability to provide computer network connections to many remote sites. During the implementation of the Washington Heights and Inwood Community Health Management Information System (WHICHIS) at the Columbia-Presbyterian Medical Center (CPMC), a number of remote connectivity issues have been encountered. Both technical and non-technical issues were significant during the installation. We developed a work-flow model for this process which may be helpful to any health care institution attempting to provide seamless remote connectivity. This model is presented and implementation lessons are discussed. PMID- 9357644 TI - Patient-Centered Access to Secure Systems Online (PCASSO): a secure approach to clinical data access via the World Wide Web. AB - The Internet's World-Wide Web (WWW) provides an appealing medium for the communication of health related information due to its ease of use and growing popularity. But current technologies for communicating data between WWW clients and servers are systematically vulnerable to certain types of security threats. Prominent among these threats are "Trojan horse" programs running on client workstations, which perform some useful and known function for a user, while breaching security via background functions that are not apparent to the user. The Patient-Centered Access to Secure Systems Online (PCASSO) project of SAIC and UCSD is a research, development and evaluation project to exploit state-of-the art security and WWW technology for health care. PCASSO is designed to provide secure access to clinical data for healthcare providers and their patients using the Internet. PCASSO will be evaluated for both safety and effectiveness, and may provide a model for secure communications via public data networks. PMID- 9357646 TI - Virtual consolidation of Boston's Beth Israel and New England Deaconess Hospitals via the World Wide Web. AB - With the advent of Integrated Healthcare Delivery Systems, medical records are increasingly distributed across multiple institutions. Timely access to these medical records is a critical need for healthcare providers. The CareWeb project provides an architecture for World Wide Web-based retrieval of electronic medical records from heterogeneous data sources. Using Health Level 7 (HL7), web technologies and readily available software components, we consolidated the electronic records of Boston's Beth Israel and Deaconess Hospitals. We report on the creation of CareWeb (freya.bidmc.harvard.edu/careweb.htm) and propose it as a means to electronically link Integrated Health Care Delivery Systems and geographically distant information resources. PMID- 9357645 TI - A strategy for the development of secure telemedicine applications. AB - Healthcare applications based on computer-supported collaboration technologies have the potential to improve the quality of care delivered to patients. Such applications can help overcome barriers to quality healthcare in the small, scattered populations of rural areas enabling telemedicine to be a part of the practice of medicine. However the growing concern about the potential for abuse through disclosure of personal health information to unauthorized parties has restricted the deployment and adoption of these potentially valuable tools. The authors, who built ARTEMIS--an Intranet healthcare collaboration facility, now describe their approach to develop secure telemedicine applications for rural healthcare practitioners. PMID- 9357647 TI - Clinical communication among health providers and systems using Web tools. AB - Three needs have driven the development of a Web front end to our legacy system. 1) A Web Intranet is needed to provide service for the quantity and diversity of platforms within our health care system. 2) Information transfer in our system is required in more than one format: in viewer-friendly, HTML format and in a database-friendly, down-loadable format. 3) The system encounters the need to electronically exchange information with providers that are not employees of our health care enterprise. This presents a problem with the authentication aspect of security for which we have devised a system to allow the carefully-monitored exchange of records with care providers who are "strangers" to our system. PMID- 9357648 TI - The PartnerWeb Project: a component-based approach to enterprise-wide information integration and dissemination. AB - A component-based health information resource, delivered on an intranet and the Internet, utilizing World Wide Web (WWW) technology, has been built to meet the needs of a large integrated delivery network (IDN). Called PartnerWeb, this resource is intended to provide a variety of health care and reference information to both practitioners and consumers/patients. The initial target audience has been providers. Content management for the numerous departments, divisions, and other organizational entities within the IDN is accomplished by a distributed authoring and editing environment. Structured entry using a set of form tools into databases facilitates consistency of information presentation, while empowering designated authors and editors in the various entities to be responsible for their own materials, but not requiring them to be technically skilled. Each form tool manages an encapsulated component. The output of each component can be a dynamically generated display on WWW platforms, or an appropriate interface to other presentation environments. The PartnerWeb project lays the foundation for both an internal and external communication infrastructure for the enterprise that can facilitate information dissemination. PMID- 9357649 TI - The importance of Java and CORBA in medicine. AB - One of the most powerful tools available for telemedicine is a multimedia medical record accessible over a wide area and simultaneously editable by multiple physicians. The ability to do this through an intuitive interface linking multiple distributed data repositories while maintaining full data integrity is a fundamental enabling technology in healthcare. We discuss the role of distributed object technology using Java and CORBA in providing this capability including an example of such a system (TeleMed) which can be accessed through the World Wide Web. Issues of security, scalability, data integrity, and usability are emphasized. PMID- 9357650 TI - A virtual repository approach to clinical and utilization studies: application in mammography as alternative to a national database. AB - A national mammography database was proposed, based on a centralized architecture for collecting, monitoring, and auditing mammography data. We have developed an alternative architecture relying on Internet-based distributed queries to heterogeneous databases. This architecture creates a "virtual repository", or a federated database which is constructed dynamically, for each query and makes use of data available in legacy systems. It allows the construction of custom tailored databases at individual sites that can serve the dual purposes of providing data (a) to researchers through a common mammography repository and (b) to clinicians and administrators at participating institutions. We implemented this architecture in a prototype system at the Brigham and Women's Hospital to show its feasibility. Common queries are translated dynamically into database specific queries, and the results are aggregated for immediate display or download by the user. Data reside in two different databases and consist of structured mammography reports, coded per BIRADS Standardized Mammography Lexicon, as well as pathology results. We prospectively collected data on 213 patients, and showed that our system can perform distributed queries effectively. We also implemented graphical exploratory analysis tools to allow visualization of results. Our findings indicate that the architecture is not only feasible, but also flexible and scaleable, constituting a good alternative to a national mammography database. PMID- 9357651 TI - HOLON: a Web-based framework for fostering guideline applications. AB - HOLON is a research and development effort in extending middleware in the healthcare field to support application development, in general, and guideline applications, in particular. This framework makes use of open standards for architecture, software, guideline KBs, clinical repository models, information encodings, and intelligent system modules and agents. By pursuing the use of such standards in our middleware components, we hope eventually to maximize reusability of the HOLON framework by others who also adhere to these open standards. This research reflects lessons learned about the extensions needed in these standards if healthcare middleware frameworks are to transparently support application developers and their users over the web. PMID- 9357652 TI - Telemedicine for AIDS patients accommodations. AB - People suffering from AIDS are subject to frequent hospitalisations. In some cases, they cannot go back home after hospitalisations, due to severe illness, family or sociologic problems. This is the reason why some therapeutic flats are at their disposal to make easier their medical follow-up after the hospital's discharge. In these Therapy Accommodation, they are treated by trained GP who often suffer from lack of information and lack of expertise in difficult cases. For this purpose we included these flats in the regional Telemedicine AIDS network to give these physicians free access to the computerised multimedia medical record of their patients and to provide them with synchronous co operation facilities. PMID- 9357653 TI - Mobile Telemedicine Testbed. PMID- 9357655 TI - Supporting shared care for diabetes patients. The synapses solution. AB - In this paper we discuss the construction of a Federated Health Care Record server within the context of the European R&D project Synapses. We describe the system using the five ODP viewpoints. From an analysis of the business process to be supported by the distributed system (the shared care for diabetes patients) requirements for the server are derived. PMID- 9357654 TI - A decentralized, community-based design for statewide immunization registries in Minnesota. AB - Incomplete immunization records and an increasingly complex immunization schedule make it difficult for parents and providers to know what shots their children or clients need. Complete and accurate immunization records are needed for day care, sports, camp, and school, but this is difficult--especially when previous immunizations have been received at different clinics. Population-based immunization registries help make complete and accurate records more easily available to parents and health care providers. Registries foster the timely sending of reminder notices for children who are due for immunizations and make it possible for providers to quickly assess immunization rates in their clinic. Public health officials use registries to determine immunization rates, to identify pockets of need where immunization rates are low and to target resources. In Minnesota, over 85% of immunizations are delivered in the private sector. Minnesota is also extensively covered by managed care organizations with an estimated 75% of the total population enrolled in some type of managed care. Strong local community public health agencies in each county also drive local solutions to community needs. These factors and others led to a de-centralized approach to the implementation of registries. The "Minnesota Model" is based on the development of community-based registries which link together local clinics, hospitals, health plans, public health departments, and schools in each region. Each community-based registry is designed to link to a state hub. This decentralized open architecture design is based on standards for data, not hardware or software. The building begins, not by implementing a state registry into which all immunizations are entered, but at the community level. Currently, 38% of Minnesota counties (representing 52% of statewide births) are involved in implementing a community-based registry, and 53% (representing 43% of statewide births) have initiated discussions with private providers. Only 9% of counties (5% of statewide births) have no current registry activity. This paper describes the steps which have been taken towards developing a decentralized statewide immunization information system for Minnesota, based on recommendations put forth by The State Immunization Practices Task Force Work Group on Immunization Registries. PMID- 9357657 TI - Desktop teleradiology in support of rural orthopedic trauma care. AB - Research has shown that diagnostic quality images for most teleradiology applications requires a sophisticated telemedicine system and access to a large amount of bandwidth. While the ideal standards have been set by those involved in evaluating teleradiology, these standards are impractical for many small rural health centers which deliver routine trauma care. While there is no disagreement about the ultimate need for this level of teleradiology support, the purpose of this research was to determine whether Orthopedists would be able to read plain radiographs of orthopedic trauma injuries using a desktop teleradiology system in support of rural trauma care. METHOD: Two radiology residents and two orthopedic residents viewed forty radiographs, twenty through a desktop teleradiology system and twenty in person. Diagnostic findings and certainty of diagnosis were recorded. FINDINGS: There was no statistically significant difference between modalities in orthopedic residents' ability to correctly diagnose orthopedic trauma injuries. Further, for those instances when the diagnosis was imprecise, the residents were aware of their inability to make an accurate diagnosis. CONCLUSION: Although the study was relatively limited and further research needs to be done, the use of desktop teleradiology in support of rural orthopedic trauma consultation is a promising alternative to the more expensive forms of telemedicine technology. PMID- 9357656 TI - Efficacy of tele-endoscopy in a rural capitated market. AB - PURPOSE: To attempt to quantify the potential for success of tele-endoscopy as a component of the VTMEDNET Plus telemedicine implementation, a multi-part prospective study was undertaken by faculty of the Vermont Initiative for Rural Health Informatics and Telemedicine. METHOD: The study was comprised of three separate parts, evaluation of image quality, cost analysis, and identification of referring providers needs and attitudes regarding tele-endoscopy. FINDINGS: The image quality was satisfactory to support remote diagnosis in most cases; there was significant cost savings in a managed care environment; referring providers were generally positive about the attributes of tele-endoscopy. CONCLUSION: Tele endoscopy is a viable and cost-effective component within a telemedicine system. PMID- 9357658 TI - Evaluation of clinical data by remote observation in trauma. PMID- 9357659 TI - Telematics in the neonatal ICU and beyond: improving care for high-risk newborns and their families. AB - The Beth Israel-Deaconess has recently been awarded one of 19 contracts from the National Library of Medicine (NLM) to develop, implement and test a telemedicine application to support the care of Very Low Birth Weight Infants. This project is the only one to focus on the care of newborns. We believe that this project will provide a new national approach to managing the care of high-risk newborns by leveraging evolving communication technology. PMID- 9357661 TI - Documenting the information content of images. AB - A standards-based message and terminology architecture has been specified to enable large-scale open and non-proprietary interchange of imaging-procedure descriptions and image-interpretation reports providing semantically-rich linkage of linguistic and non-linguistic information. The DICOM Structured Reporting Supplement, now available for trial use, embodies this interdependent message/terminology architecture. A DICOM structured report object is a self describing information structure that can be tailored to support diverse clinical observation reporting applications by utilization of templates and context dependent terminology from an external message/terminology mapping resource such as the SNOMED DICOM Microglossary (SDM), HL7 Vocabulary, or Terminology Resource for Message Standards (TeRMS). PMID- 9357660 TI - Application of UMLS indexing systems to a WWW-based tool for indexing of digital images. PMID- 9357662 TI - Evaluation of a visualization-based approach to functional brain mapping. AB - We describe a method for mapping stimulation data, obtained at the time of neurosurgery for intractable epilepsy, onto a 3D MRI-based neuroanatomic model of the individual patient. The mapping is done by comparing an intraoperative photograph of the exposed cortical surface with a computer-based MR visualization of the surface, interactively indicating corresponding stimulation sites, and recording 3-D MR machine coordinates of the indicated sites. Repeatability studies were performed to validate the accuracy of the mapping technique. Six observers--a neurosurgeon, a radiologist, and four computer scientists, independently mapped 218 stimulation sites from 12 patients. The mean distance of the six locations from the mean location of each site was 2.07 mm, with a standard deviation of 1.5 mm, or within 5.07 mm with 95% confidence. Since the surgical sites are accurate within approximately 1 cm, these results show that the visualization-based approach is accurate within the limits of the stimulation maps. When incorporated within the kind of information system envisioned by the Human Brain Project, this anatomically-based method will not only provide a key link between non-invasive and invasive approaches to understanding language organization, but will also provide the basis for studying the relationship between language function and anatomical variability. PMID- 9357663 TI - Representation of the Gastrointestinal Endoscopy Minimal Standard Terminology in the SNOMED DICOM microglossary. AB - In Gastroenterology, endoscopic images and interpretation reports are essential elements of the patient record. The Digital Imaging and Communications in Medicine (DICOM) Visible Light and Structured Reporting Standards provide a standard representation of images and reports. However, the message standards are not sufficient in themselves. Controlled terminology is needed to enable interchange of patient records and to facilitate the pooling of multi-center data for large-scale outcomes studies and clinical research. The ASGE has joined with European and Japanese colleagues to develop and publish a lexicon of endoscopic terminology. The lexicon is being tested now in a multi-center trial. In addition, the ASGE is collaborating with the DICOM Standards Committee to transform the endoscopic lexicon into a database structure that is suitable for use with the DICOM Visible Light and Structured Reporting Standards. The combination of an internationally accepted, tested and non-proprietary lexical standard and a DICOM message standard supporting endoscopic images and reports represents a powerful tool for clinicians to improve communication, research and the quality of care. PMID- 9357664 TI - Critical success factors for a hospital-wide PACS. AB - Since 1996 a picture archiving and communications system (PACS) is installed at the university hospital of Freiburg. The PACS is integrated in the hospital information system (HIS) and several modalities of different vendors are attached to it. During the implementation phase three critical factors to the success of our PACS installation where identified: the support of the workflow, the interface of the radiological information system (RIS) to the modalities, and the security policy to allow hospital-wide access to the images and results in the PACS. PMID- 9357665 TI - Acceptability and usage patterns of an image analysis workstation. AB - Critical to the successful deployment and use of new computer systems is the acceptance of the system by the users, i.e., the clinicians. We describe a study which evaluated, in an experimental setting, the potential acceptability of an image analysis workstation for radiation therapy. The acceptability and usage patterns were measured using semi-structured questionnaires and maintaining logs of user interactions. The results of the study showed that the radiation oncologists, who were the subjects for the study, perceived the workstation as acceptable. The results also suggested several areas for improvement of workstation that could increase its acceptance in the clinical setting. PMID- 9357667 TI - Managing medical research data with a Web-Interfacing Repository Manager. AB - This paper describes the Web-Interfacing Repository Manager (WIRM), a perl toolkit for managing and deploying multimedia data, which is built entirely from free, platform-independent components. The WIRM consists of an object-relational API layered over a relational database, with built-in support for file management and CGI programming. The basic underlying data structure for all WIRM data is the repository object, a perl associative array whose values are bound to a row of a table in the relational database. Based on our experience implementing a target application (the Brain Mapper Console), we describe five stages through which a system passes as it evolves from a primitive file hierarchy to a full-fledged repository console. PMID- 9357666 TI - The Virtual Microscope. AB - We present the design of the Virtual Microscope, a software system employing a client/server architecture to provide a realistic emulation of a high power light microscope. We discuss several technical challenges related to providing the performance necessary to achieve rapid response time, mainly in dealing with the enormous amounts of data (tens to hundreds of gigabytes per slide) that must be retrieved from secondary storage and processed. To effectively implement the data server, the system design relies on the computational power and high I/O throughput available from an appropriately configured parallel computer. PMID- 9357668 TI - Automated measurement of retinal vascular tortuosity. AB - Automatic measurement of blood vessel tortuosity is a useful capability for automatic ophthalmological diagnostic tools. We describe a suite of automated tortuosity measures for blood vessel segments extracted from RGB retinal images. The tortuosity measures were evaluated in two classification tasks: (1) classifying the tortuosity of blood vessel segments and (2) classifying the tortuosity of blood vessel networks. These tortuosity measures were able to achieve a classification rate of 91% for the first problem and 95% on the second problem, which confirms that they capture much of the ophthalmologists' notion of tortuosity. PMID- 9357670 TI - Using 3-D shape models to guide segmentation of MR brain images. AB - Accurate segmentation of medical images poses one of the major challenges in computer vision. Approaches that rely solely on intensity information frequently fail because similar intensity values appear in multiple structures. This paper presents a method for using shape knowledge to guide the segmentation process, applying it to the task of finding the surface of the brain. A 3-D model that includes local shape constraints is fitted to an MR volume dataset. The resulting low-resolution surface is used to mask out regions far from the cortical surface, enabling an isosurface extraction algorithm to isolate a more detailed surface boundary. The surfaces generated by this technique are comparable to those achieved by other methods, without requiring user adjustment of a large number of ad hoc parameters. PMID- 9357669 TI - Modeling mechanical cardiopulmonary interactions for virtual environments. AB - We have developed a computer system for modeling mechanical cardiopulmonary behavior in an interactive, 3D virtual environment. The system consists of a compact, scalar description of cardiopulmonary mechanics, with an emphasis on respiratory mechanics, that drives deformable 3D anatomy to simulate mechanical behaviors of and interactions between physiological systems. Such an environment can be used to facilitate exploration of cardiopulmonary physiology, particularly in situations that are difficult to reproduce clinically. We integrate 3D deformable body dynamics with new, formal models of (scalar) cardiorespiratory physiology, associating the scalar physiological variables and parameters with corresponding 3D anatomy. Our approach is amenable to modeling patient-specific circumstances in two ways. First, using CT scan data, we apply semi-automatic methods for extracting and reconstructing the anatomy to use in our simulations. Second, our scalar models are defined in terms of clinically-measurable, patient specific parameters. This paper describes our approach and presents a sample of results showing normal breathing and acute effects of pneumothoraces. PMID- 9357671 TI - Automatic detection of cardiac contours on MR images using fuzzy logic and dynamic programming. AB - This paper deals with the use of fuzzy logic and dynamic programming in the detection of cardiac contours in MR Images. The definition of two parameters for each pixel allows the construction of the fuzzy set of the cardiac contour points. The first parameter takes into account the grey level, and the second the presence of an edge. A corresponding fuzzy matrix is derived from the initial image. Finally, a dynamic programming with graph searching is performed on this fuzzy matrix. The method has been tested on several MR images and the results of the contouring were validated by an expert in the domain. This preliminary work clearly demonstrates the interest of this method, although a formal evaluation has to be done. PMID- 9357672 TI - Dynamic organization of search results using the UMLS. AB - When people search the medical literature, they often are overwhelmed by the large number of documents retrieved. Many systems try to solve this problem by helping the user formulate a more specific search strategy. However, when users do not have a more specific question, they need tools to help them explore and understand the results, rather than to eliminate a portion of those results. This paper describes an approach that addresses this need by automatically grouping the results of a broad search into meaningful categories based on the user's query. This approach combines the main benefit of clustering techniques with the main benefit of classification techniques by taking advantage of the domain knowledge present in the UMLS. I present a preliminary evaluation that demonstrates that a categorization produced by this approach corresponds reasonably well to a physician's categorization. PMID- 9357673 TI - Query expansion using the UMLS Metathesaurus. AB - Recent work has demonstrated the importance of query expansion for improving retrieval effectiveness when applying statistically-based systems to MEDLINE citations. The research has suggested the use of retrieval feedback for enhancing the original text of users' queries. As an alternative method of query expansion, we propose the use of the MetaMap program for associating UMLS Metathesaurus concepts with the original query. Our experiments show that query expansion based on MetaMap compares favorably with retrieval feedback. We conclude that the optimal strategy would be to combine the two techniques. PMID- 9357674 TI - MedWeaver: integrating decision support, literature searching, and Web exploration using the UMLS Metathesaurus. AB - Integrating functions from disparate and widely-distributed information systems has been an interest of the medical informatics community for some time. Barriers to progress have included the lack of network-accessible information sources, inadequate methods for inter-system messaging, and lack of vocabulary translation services. With the advent of the World Wide Web (WWW) and the evolution of the National Library of Medicine's Unified Medical Language System (UMLS), it is now possible to develop applications that integrate functions from diverse, distributed systems. In this paper we describe one such system, MedWeaver, a WWW application that integrates functions from a decision support application (DXplain), a literature searching system (WebMedline), and a clinical Web searching system (CliniWeb) using the UMLS Metathesaurus for vocabulary translation. This system demonstrates how application developers can design systems around anticipated clinical information needs and then draw together the needed content and functionality from diverse sources. PMID- 9357675 TI - A UMLS-based method for integrating information databases into an Intranet. AB - The Internet and the World Wide Web provide today end-users with capabilities to access universally to information in various and heterogeneous databases. The biomedical domain benefits from this new technology, specially for information retrieval by searching and browsing various sites. Nevertheless, end-users may be disoriented by specific ways to access information on different servers. In the framework of an Intranet design and development, we present a method for integrating information databases based on knowledge sources of the UMLS. The method provides designers of a Web site with facilities to implement an easy and homogeneous access to information. The pages are built dynamically and displayed according to a style sheet and their content stored in a database during the design phase. The database also describes the links between pages. Moreover, this organization provides administrators with powerful capabilities to manage Web sites. PMID- 9357676 TI - Standardized problem list generation, utilizing the Mayo canonical vocabulary embedded within the Unified Medical Language System. AB - VOCABULARY: The Mayo problem list vocabulary is a clinically derived lexicon created from the entries made to the Mayo Clinic's Master Sheet Index and the problem list entries made to the Impression/ Report/Plan section of the Clinical Notes System over the last three years. The vocabulary was reduced by eliminating repetition including lexical variants, spelling errors, and qualifiers (Administrative or Operational terms). Qualifiers are re-coordinated with other terms, at run-time, which greatly increased the number of input strings which our system is capable of recognizing. IMPLEMENTATION: The Problem Manager is implemented using standard windows tools in a Windows NT environment. The interface is designed using Object Pascal. HTTP calls are passed over the World Wide Web to a UNIX based vocabulary server. The server returns a document, which is read into Object Pascal structures, parsed, filtered and displayed. STUDY: This paper reports the results of a recent Usability Trial focused on assessing the viability of this mechanism for standardized problem entry. Eight clinicians engaged in eleven scenarios and responded as to their satisfaction with the systems performance. These responses were observed, videotaped and tabulated. Clinicians in this study were able to find acceptable diagnoses in 91.1% of the scenarios. The response time was acceptable in 92.5% of the scenarios. The presentation of related terms was stated to be useful in at least one scenario by seven of the eight participants. All clinicians wanted to make use of shortcuts which would minimize the amount of typing necessary to encode the concept they were searching for (e.g. Abbreviations, Word Completion). CONCLUSIONS: Clinicians are willing to choose a canonical term from a suggested list (as opposed to their own wording). Clinicians want an "intelligent" system, which would suggest terms within a category (e.g. Types of "Migraine"). They are able to make functional use of our system, in its current state of development. Finally, all clinicians appreciate the value of encoding their problems in a standardized vocabulary, toward improved research, education and practice. PMID- 9357677 TI - The Stanford Health Information Network for Education: integrated information for decision making and learning. AB - Although multiple decision support systems have been built for physicians, efficient delivery of valid and complete medical knowledge remains an elusive goal. In this paper we describe a new project, the Stanford Health Information Network for Education (SHINE). SHINE unifies core medical resources in an intuitive interface to support clinical decision making. Included in the description is a novel paradigm for continuing medical education (CME). PMID- 9357679 TI - Comparing SNOMED and ICPC retrieval accuracies using relational database models. AB - While SNOMED International has been generally accepted by the international community of pathologists, its use for primary and secondary care remains limited. This can probably be attributed to the coding complexity of clinical concepts into this multiaxial postcoordinated nomenclature. The SNOMED editors propose the use of multiple codes (aggregates) for any nuanced clinical concept, thus allowing alternative rigorous representations of the concept with SNOMED codes. Some classification critics argue whether such redundant coding precludes precise retrieval of data. This research was initiated to compare the retrieval accuracies of a relational database using a simplified model of SNOMED against a classification-based model. SNOMED-based queries showed improvement over ICPC based queries, regardless of the use of SNOMED cross-references. The addition of the latter significantly improved the queries sensitivity and false negative rate. In conclusion, the authors recommend using aggregates of SNOMED codes in relational database designs over classification-based designs in order to improve retrieval accuracy. PMID- 9357678 TI - Puya: a method of attracting attention to relevant physical findings. AB - Puya is a method that compares the physical exam in an electronic clinical note with a set of stereotypical physical exam sentences that have been previously classified as "normal". The note is then displayed in a web browser with normal findings clearly delineated. The list of stereotypical sentences comes from a set of physical findings found within extensive electronic medical record. This list is then screened to select only those that represent "normal" findings, a process that yields 96% total agreement among 4 clinicians surveyed. This final list of stereotypical "normal" sentences accounts for 64% of the clinical narrative text. Sentences in the clinical note that do not match sentences in the "normal" list are assumed to be "abnormal". Puya screened 98 clinical notes consisting of 610 individual sentences. Puya achieved a sensitivity of 100%, a specificity of 63%, a positive predictive value of 44% and a negative predictive value of 100%. This leads to an application that reduces informational noise. PMID- 9357680 TI - Conceptual mapping of user's queries to medical subject headings. AB - This paper describes a way to map users' queries to relevant Medical Subject Headings (MeSH terms) used by the National Library of Medicine to index the biomedical literature. The method, called SENSE (SEarch with New SEmantics), transforms words and phrases in the users' queries into primary conceptual components and compares these components with those of the MeSH vocabulary. Similar to the way in which most numbers can be split into numerical factors and expressed as their product--for example, 42 can be expressed as 2*21, 6*7, 3*14, 2*3*7,--so most medical concepts can be split into "semantic factors" and expressed as their juxtaposition. Note that if we split 42 into its primary factors, the breakdown is unique: 2*3*7. Similarly, when we split medical concepts into their "primary semantic factors" the breakdown is also unique. For example, the MeSH term 'renovascular hypertension' can be split morphologically into reno, vascular, hyper, and tension--morphemes that can then be translated into their primary semantic factors--kidney, blood vessel, high, and pressure. By "factoring" each MeSH term in this way, and by similarly factoring the user's query, we can match query to MeSH term by searching for combinations of common factors. Unlike UMLS and other methods that match at the level of words or phrases, SENSE matches at the level of concepts; in this way, a wide variety of words and phrases that have the same meaning produce the same match. Now used in PaperChase, the method is surprisingly powerful in matching users' queries to Medical Subject Headings. PMID- 9357681 TI - Virtual shelves. II: A unified catalog for a heterogeneous collection. AB - We describe a unified catalog of traditional and digital resources in medical informatics, using standard cataloging principles (AACR2), schemes (LCC), and coding formats (USMARC). The unified catalog integrates the bibliographic records of physical items in the heterogeneous collection with the bibliographic records of network accessible digital items, using prescribed cataloging formats to new effect. The unified catalog is collections-based. We do not use the MARC 856 field to specify the network location of a digital item. The location of a digital item is determined by mapping its call number through a location guide to a network address. This mapping is strictly isomorphic to the way the shelf location of a physical item is determined within the bounds of a controlled collection. PMID- 9357682 TI - Supporting infobuttons with terminological knowledge. AB - We have developed several prototype applications which integrate clinical systems with on-line information resources by using patient data to drive queries in response to user information needs. We refer to these collectively as infobuttons because they are evoked with a minimum of keyboard entry. We make use of knowledge in our terminology, the Medical Entities Dictionary (MED) to assist with the selection of appropriate queries and resources, as well as the translation of patient data to forms recognized by the resources. This paper describes the kinds of knowledge in the MED, including literal attributes, hierarchical links and other semantic links, and how this knowledge is used in system integration. PMID- 9357683 TI - A method for representing search results in three dimensions. AB - This paper presents a new method for representing results of an information retrieval search in a three dimensional environment. Aside from the fact that users find 3-D interfaces visually appealing, there are strong practical reasons for developing 3-D representations of search results. Traditional information retrieval systems present results in ordered lists which are difficult to browse, and exclude useful information. The current method employs a multivariate statistical method called Local Latent Semantic Indexing (LLSI) to create meaningful local dimensions in which to view search results. A prototype Internet ready system is described which utilizes Virtual Reality Modeling Language (VRML) to display search results. Preliminary tests of this system with a small collection of MEDLINE articles are very encouraging. PMID- 9357685 TI - Internet access to a medical case repository for teaching and analysis. AB - The acute radiation syndrome is a rare disease. Teaching its treatment and the achievement of scientific progress require a sufficient number of accessible case histories. An information system for standardized recording of acute radiation syndrome case histories has been established in the framework of the World Health Organization Radiation Medical Emergency Preparedness and Assistance Network Centers. The information system has been realized in client/server architecture. The centers are able to access the database by easy to use graphical user interfaces via the Internet. Focusing on optimization of network traffic and flexible information retrieval the hypertext transfer protocol has not been applied. At the moment the system is tested on the campus of the University of Ulm before it will be installed at the other centers. PMID- 9357684 TI - A Java-based multi-institutional medical information retrieval system. AB - JAMI (Java-based Agglutination of Medical Information) is designed as a framework for integrating heterogeneous information systems used in healthcare related institutions. It is one of the implementations under the W3-EMRS project 1 aimed at using the World Wide Web (Web) to unify different hospital information systems. JAMI inherited several design decisions from the first W3-EMRS implementation described in, including using the Web as the communication infrastructure and HL7 as the communication protocol between the heterogeneous systems and the W3-EMRS systems. In addition, JAMI incorporates the growing Java technologies and has a more flexible and efficient architecture. This paper describes JAMI's architecture and implementation. It also present two instances of JAMI, one for the integration of different hospital information systems and another for the integration of two heterogeneous systems within a single hospital. Some important issues for the further development of JAMI, including security and confidentiality, data input and decision support are discussed. PMID- 9357686 TI - The Healthcare Administrator's Associate: an experiment in distributed healthcare information systems. AB - The Healthcare Administrator's Associate is a collection of portable tools designed to support analysis of data retrieved via the Internet from diverse distributed healthcare information systems by means of the InfoSleuth system of distributed software agents. Development of these tools is part of an effort to enhance access to diverse and geographically distributed healthcare data in order to improve the basis upon which administrative and clinical decisions are made. PMID- 9357687 TI - Linking a clinical system to heterogeneous information resources. AB - We present a model for providing clinical information system (CIS) users with quick access to high quality information resources. We have developed a chest X ray information button application which is attached to the chest X-ray reports in the CIS at the Columbia Presbyterian Medical Center. The application generates questions based on clinical information, user interest, generic question templates, and resource availability. It then provides answers to the questions through integrated access to heterogeneous information resources including the CIS itself and publicly accessible Web resources. PMID- 9357688 TI - Conducting the NLM/AHCPR Large Scale Vocabulary Test: a distributed Internet based experiment. AB - The Large Scale Vocabulary Test, sponsored by the National Library of Medicine (NLM) and the Agency for Health Care Policy and Research (AHCPR), was conducted to determine the extent to which a combination of existing health-related terminologies cover vocabulary needed in health care information systems. The test was conducted over the Internet using a sophisticated World Wide Web interface with over 60 participants and over 40,000 terms submitted. This paper discusses the issues encountered in the design and execution of the experiment, including the design of the interface and the issues of recruitment, training, and guidance of remote participants over the Internet. Test data are currently undergoing expert review. Upon completion of the expert review, the results of the test will be fully reported. PMID- 9357690 TI - A clinically derived terminology: qualification to reduction. AB - Mayo Foundation is developing synonym rich entry points for the recording of patient problems by clinicians, which will map to the KP-Mayo Convergent Medical Terminology. We describe the empirical sources for these terminology components, and how the number and complexity of the terms could be substantially reduced by the introduction of a Qualifier axis. The expressive power of these entry points is dramatically enhanced by this axis. This work is being integrated into terminology navigation modules being jointly developed with Lexical Technology, which leverages UMLS content. It will from the basis for structured problem entry into Mayo's Computer-based Electronic Record. PMID- 9357691 TI - WWW-available conceptual integration of medical terminologies: the ONIONS experience. AB - We present the most applicable aspects of our research in the conceptual integration of terminologies. From past experience, we claim that the conceptualizations provided for terminological ontologies need to be philosophically and linguistically grounded. We developed ONIONS, a methodology for integrating domain terminologies by exploiting a library of generic ontologies. Our current focus is on flexible and cooperative modelling of terminological ontologies. We adopt modular and negotiable architectures of ontologies and some WWW-oriented tools, such as Ontolingua and Ontosaurus. PMID- 9357689 TI - Department of Veterans Affairs, University of Utah consortium participation in the NLM/AHCPR Large Scale Vocabulary Test. AB - The Large Scale Vocabulary Test (LSVT) was designed to evaluate how well the Metathesaurus plus planned additions to Meta covered the documentation needs of clinicians. Our consortium collected 10,538 clinical narratives from patient problem lists recorded at 65 Veterans Hospitals, internal medicine ambulatory care practices, diagnostic history and physical examination data elements from Iliad, and nursing shift notes and emergency transport patient records. The results showed 94% of submitted terms resulted in acceptable matches. 49% of submitted terms were judged to be synonymous with the match terms, 35% were judged to be more specific (usually due to modifiers), 2%, were less specific, and 6% had an associative relationship. In 8% of cases either no match was found by the LSVT interface or all proposed matches were rejected by the raters. The LSVT content was quite suitable for coding our narratives. Necessary improvements for an electronic record would include the ability to compose modifiers together with root concepts. PMID- 9357692 TI - Assessing the feasibility of large-scale natural language processing in a corpus of ordinary medical records: a lexical analysis. AB - OBJECTIVE: Identify the lexical content of a large corpus of ordinary medical records to assess the feasibility of large-scale natural language processing. METHODS: A corpus of 560 megabytes of medical record text from an academic medical center was broken into individual words and compared with the words in six medical vocabularies, a common word list, and a database of patient names. Unrecognized words were assessed for algorithmic and contextual approaches to identifying more words, while the remainder were analyzed for spelling correctness. RESULTS: About 60% of the words occurred in the medical vocabularies, common word list, or names database. Of the remainder, one-third were recognizable by other means. Of the remaining unrecognizable words, over three-fourths represented correctly spelled real words and the rest were misspellings. CONCLUSIONS: Large-scale generalized natural language processing methods for the medical record will require expansion of existing vocabularies, spelling error correction, and other algorithmic approaches to map words into those from clinical vocabularies. PMID- 9357693 TI - Corpus-based identification and refinement of semantic classes. AB - Medical Language Processing (MLP), especially in specific domains, requires fine grained semantic lexica. We examine whether robust natural language processing tools used on a representative corpus of a domain help in building and refining a semantic categorization. We test this hypothesis with ZELLIG, a corpus analysis tool. The first clusters we obtain are consistent with a model of the domain, as found in the SNOMED nomenclature. They correspond to coarse-grained semantic categories, but isolate as well lexical idiosyncrasies belonging to the clinical sub-language. Moreover, they help categorize additional words. PMID- 9357694 TI - Evaluating a normalized conceptual representation produced from natural language patient discharge summaries. AB - The Menelas project aimed to produce a normalized conceptual representation from natural language patient discharge summaries. Because of the complex and detailed nature of conceptual representations, evaluating the quality of output of such a system is difficult. We present the method designed to measure the quality of Menelas output, and its application to the state of the French Menelas prototype as of the end of the project. We examine this method in the framework recently proposed by Friedman and Hripcsak. We also propose two conditions which enable to reduce the evaluation preparation workload. PMID- 9357695 TI - Towards a comprehensive medical language processing system: methods and issues. AB - Natural language processing (NLP) systems can help solve the data entry problem by providing coded data from textual reports for clinical applications. A number of NLP systems have shown promise, but have not yet achieved wide-spread use for practical applications. In order to achieve such use, a system must have broad coverage of the clinical domain and not be restricted to limited applications. In addition, an NLP system must perform satisfactorily for real-world applications. This paper describes methods and issues associated with an ongoing extension of MedLEE, an operational NLP system, from a limited domain to a domain that encompasses comprehensive clinical information. PMID- 9357696 TI - Maintaining the integrity of a Web-based medical vocabulary glossary. AB - Medical vocabulary databases are vital components of electronic medical record (EMR) systems. While their performance and efficiency has been extensively explored by many authors, few have dealt with the maintenance of their semantic integrity. Clinicians' preference for an optimistic system has introduced a need for monitoring and filtering proposed additions to the vocabulary tables. We propose the use of batch processing and memo-posting as means of implementing a World-Wide-Web-based Controlled Vocabulary Glossary as a monitored optimistic system. PMID- 9357697 TI - Development of a change model for a controlled medical vocabulary. AB - Managing change in controlled medical vocabularies is labor intensive and costly, but change is inevitable if vocabularies are to be kept up to date. The changes that are appropriate for a controlled medical vocabulary depend on the data stored for that vocabulary, and those data in turn depend on the needs of users. The set of change operations is the change model; the data stored about concepts comprise the concept model. Because the change model depends directly on the concept model, a discussion of the former necessitates a discussion of the latter. In this paper, we first present a set of tasks that we believe controlled medical vocabularies should handle. Next, we describe our concept model for a controlled medical vocabulary. Then, we review the literature on changes in existing vocabulary systems. Finally, we present our change model. We call our system, which incorporates the concept model and change model, the General Online Dictionary of Medicine (GOLDMINE). PMID- 9357698 TI - Terminological systems: bridging the generation gap. AB - A rigorous formal description of the intended behaviour of a compositional terminology, a 'third generation' system, enables powerful semantic processing techniques to assist in the building of a large terminology. Use of an intermediate representation derived from such a formalism, but simplified to resemble a 'second generation' system, enables authors to work in an simpler and more familiar environment, avoiding many of the technical complications of the 'third generation' system. PMID- 9357699 TI - Semantic quality through semantic definition: refining the Read Codes through internal consistency. AB - Checks of internal consistency in controlled medical vocabularies facilitate their development and assist refinement of the underlying terminological model. Two simple checks of consistency between knowledge in the subtype hierarchy and that in semantic definitions of concepts are described. It is proposed that these checks are a helpful adjunct to, but not a replacement for, large-scale involvement of domain experts in construction of controlled vocabularies. PMID- 9357700 TI - Compositional and enumerative designs for medical language representation. AB - Medical language is in essence highly compositional, allowing complex information to be expressed from more elementary pieces. Embedding the expressive power of medical language into formal systems of representation is recognized in the medical informatics community as a key step towards sharing such information among medical record, decision support, and information retrieval systems. Accordingly, such representation requires managing both the expressiveness of the formalism and its computational tractability, while coping with the level of detail expected by clinical applications. These desiderata can be supported by enumerative as well as compositional approaches, as argued in this paper. These principles have been applied in recasting a frame-based system for general medical findings developed during the 1980s. The new system captures the precise meaning of a subset of over 1500 medical terms for general internal medicine identified from the Quick Medical Reference (QMR) lexicon. In order to evaluate the adequacy of this formal structure in reflecting the deep meaning of the QMR findings, a validation process was implemented. It consists of automatically rebuilding the semantic representation of the QMR findings by analyzing them through the RECIT natural language analyzer, whose semantic components have been adjusted to this frame-based model for the understanding task. PMID- 9357701 TI - Formative evaluation of a frame-based model of locative relationships in human anatomy. AB - The verb structure of narrative text in a gross anatomy textbook was analyzed to identify locative relationships. The 169 locative indicators were organized semantically to construct a frame-based model. The validity and coverage of the model was assessed and compared with the UMLS Semantic Net Relations using a novel test set of 71 indicators. All mapped directly to the frame model, while 60% mapped directly to UMLS. PMID- 9357702 TI - Partitioning a vocabulary's IS-A hierarchy into trees. AB - Controlled medical vocabularies are useful in application areas such as medical information-systems and decision-support. However, such vocabularies are large and complex, and working with them can be daunting. It is important to provide a means for orienting users to the vocabulary's contents. This paper introduces a methodology for partitioning a vocabulary into small, meaningful pieces. The partitioning is done with respect to the vocabulary's IS-A hierarchy. The methodology, based on a set of rules for refining the IS-A hierarchy, is a process carried out by a user in conjunction with the computer. The methodology is demonstrated on a complex portion of a vocabulary. PMID- 9357703 TI - SGML as a message interchange format in healthcare. AB - INTRODUCTION: In 1993, The European Committee for Standardization (CEN) studied several syntaxes for interchange formats in healthcare, but excluded SGML due to resource constraints. We sought to extend the CEN report and formally evaluate the use of SGML as a message interchange format. METHODS: We followed the methodology set forth by CEN, using their example scenarios and healthcare data model. General message descriptions based on this model set the functional requirements for the interchange format. These general requirements are then mapped into SGML to see how well they can be supported. RESULTS: Results follow the CEN format, enabling a direct comparison of SGML with ASN.1, ASTM E1238, EDIFACT, EUCLIDES, and ODA (those syntaxes studied by CEN). CONCLUSION: SGML compares favorably with other syntaxes investigated by CEN. None of the interchange formats support all functional requirements. Optimal and standard mechanisms of combining different formats through a modular approach to achieve greater overall functionality requires further study. PMID- 9357704 TI - SNOMED RT: a reference terminology for health care. AB - We describe the framework for SNOMED RT (Reference Terminology), designed to complement the broad coverage of medical concepts in SNOMED with a set of enhanced features that significantly increases its value as a reference terminology for representing clinical data. We describe what is meant by a reference terminology, and differentiate SNOMED RT from specialized terminologies that enable user interfaces, electronic messaging, or natural language processing, as well as from other specialized reference terminologies whose primary purpose is for representing data that is not primarily clinical in nature. We then describe how SNOMED RT represents multiple hierarchies and incorporates description logic. We believe that such a comprehensive set of concepts at multiple levels of granularity, with multiple logic-based subsumption hierarchies can meet the requirements of a reference terminology for health care. PMID- 9357705 TI - Evaluating the terminology requirements to support multi-disciplinary diabetes care. AB - Diabetes mellitus is a multi-system disease requiring lifetime multi-disciplinary care, which has proven individual and economic benefits. The delivery of service involves co-operation and communication between patient, carer and health care professionals, and systematic auditing of processes and outcomes. Sustained improvement necessitates regular data acquisition, aggregation and analysis. The terminology requirements to support patient-centred records and identified datasets are examined, and differences in purpose and scope highlighted. The many stakeholders involved in diabetes care have their own sublanguages and terminology requirements which need harmonising around a common core. The problems and solutions of accommodating these needs are explored in relation to the Read Thesaurus. PMID- 9357706 TI - Conceptual schemata for terminology: a continuum from headings to values in patient records and messages. AB - We developed a technique of reverse engineering to extract a conceptual schema- also called "categorial structure" in the European standard CEN ENV 12264 (MoSe)- from a set of terminological phrases. The technique was originally applied to coding systems, ie. to large value sets. We applied this technique to subsets of two new terminological resources for message standards: headings of patient record from Clinical LOINC and names of "Context Groups" for structured reporting from SNOMED DICOM Microglossary (SDM). Both sources provide context-independent names for message fields and domains of admitted values. Therefore conceptual schemata on the potential content for a field are compatible with the ones on names of the fields themselves. Both kinds of schemata can be compared and integrated with conceptual schemata for the information system that manages the patient record. This continuity in the schemata allows the coupling of applications with different organization of data, and will facilitate mapping from an application to standard messages and viceversa. Moreover, the simplified representation produced according the MoSe approach is easy to understand by healthcare operators, allowing their progressive involvement in cooperative efforts of design, discussion and validation of the schemata. PMID- 9357707 TI - Expressiveness of the Breast Imaging Reporting and Database System (BI-RADS). AB - The Breast Imaging Reporting and Database System (BI-RADS) was developed by the American College of Radiology and is used by a number of computerized mammography tracking systems. The ability of BI-RADS to encode the data contained in 300 mammography reports at the Columbia-Presbyterian Medical Center was examined. BI RADS was able to encode normal reports and "special masses" (such as lymph nodes) without difficulty. However, none of the general masses and only 17% of the calcifications could be encoded in BI-RADS. The implications of this for the design of mammography databases are discussed. PMID- 9357708 TI - Teaching skills for accessing and interpreting information from systematic reviews/meta-analyses, practice guidelines, and the Internet. AB - Skills and practice related to accessing and interpreting clinical information from systematic reviews/meta-analyses, practice guidelines, and the Internet have been integrated into a new senior year elective designed to teach medical students how to critically appraise information from a variety of sources and evaluate it's applicability to patient care. Small groups of senior medical students under the direction of a multidisciplinary team (behavioral scientist, information specialist, physician) facilitate discussions of clinical articles using checklists designed to evaluate their quality. The central feature of the course is a demonstration of the Cochrane Database of Systematic Reviews (CDSR), an electronic journal distributed by BMJ Publishing, and the requirement that students conduct a literature review on a topic of their choice and present an oral and written summary in the form of a "draft" meta-analysis. Students are provided with strategies to "surf" the Internet/WWW for information, e.g., practice guidelines/treatment protocols, descriptions of on-going clinical trials. A total of 52 students have participated to date. Students have selected project topics across a wide range of medical disciplines, including internal medicine, family practice, OB/GYN, pediatrics, surgery, neurology, emergency medicine, and psychiatry. The course is one of the most favorably evaluated of all senior electives and rated more favorably than the overall mean ratings for all electives combined on 8 of 9 scales, including "Quality of course overall" (4.39 vs. 3.92 on 5-point scale). PMID- 9357709 TI - A decision analytic method for scoring performance on computer-based patient simulations. AB - As computer based clinical case simulations become increasingly popular for training and evaluating clinicians, approaches are needed to evaluate a trainee's or examinee's solution of the simulated cases. We developed a decision analytic approach to scoring performance on computerized patient case simulations. We developed decision models for computerized patient case simulations in four specific domains in the field of infectious disease. The decision models were represented as influence diagrams. A single decision node represents the possible diagnoses the user may make. One chance node represents a probability distribution over the set of competing diagnoses in the simulations. The value node contains the utilities associated with all possible combinations of diagnosis and disease. All relevant data that the user may request from the simulation are represented as chance nodes with arcs to or from the diagnosis node and/or each other. Probabilities in the decision model were derived from the literature, where available, or expert opinion. Utilities were assessed by standard gamble from clinical experts. The process of solving computer based patient simulations involves repeated cycles of requesting data (history, physical examination or laboratory) and receiving these data from the simulations. Each time the user requests clinical data from the simulation, the influence diagram is evaluated with and without an arc from the corresponding chance node to the decision node. The difference in expected utility between the two solutions of the influence diagram represents the expected value of information (VOI) from the requested clinical datum. The ratio of the expected VOI from the data requested and the expected value of perfect information about the diagnosis is a normative measure of the quality of each of the user's data requests. This approach provides a continuous measure of the quality of the user's data requests in a way that is sensitive to the previous data collected. The score distinguishes serious from minor misdiagnoses. And the same influence diagram can be used to evaluate performance on multiple simulations in the same clinical domain. PMID- 9357710 TI - Meeting the information needs of patients: results from a patient focus group. AB - Changing roles in health care call for patients to share increased responsibility for managing their health. Patients may need additional health-related information to participate more fully in health care decisions. We examined patients' information needs from the perspective of clinicians, educational software vendors, and patients. The most instructive information came directly from patients in focus groups. The participants in our focus groups clearly sought more information about their health than they had received during visits with their physicians. Patients' needs were specific to their individual clinical situation, and timing was critical. Although physicians spend a significant amount of time on patient education during an encounter, patients typically formulate their questions after the encounter. We used the results of focus groups to develop desired characteristics of patient education material that addresses patients' information needs. Providers who understand and address these needs will be in a better position to effectively engage patients' active participation in their health care. PMID- 9357711 TI - Implementing computers in ambulatory care: implications of physician practice patterns for system design. AB - This paper presents pre-implementation data from the internal medicine division of a large physician group practice scheduled to implement an electronic medical record (EMR). Data were gathered through short-term participant observation and interviews. Findings indicate that (1) most physicians anticipate enough benefits to be willing to use the system; (2) computers must be accessible, easy to log into, and provide for physician movement and interrupted sessions; (3) many physicians are concerned about losing eye contact with patients; (4) it is unrealistic to expect even good typists to enter their own long notes; (5) staged implementation, with order entry introduced first, may help physicians adapt gradually; and (6) training should include protected time for instructional sessions for physicians, simulated patient encounters to help physicians adapt their practice patterns, and tutors available to answer questions in the clinical setting. PMID- 9357712 TI - Factors affecting the diffusion of the Computer-Based Patient Record. AB - A survey of the perceptions of 629 informatics experts representing 67 institutions with accredited schools of medicine was used to identify factors most important in implementing the Computer-Based Patient Record. A model outlined three theoretical factors: Innovation Attributes (attributes inherent in the CPR itself); Organizational Attributes; and Boundary-Spanning Attributes (related to marketing efforts). The model was explored using multiple regression techniques to delineate the relative importance of 15 variables within the three sets of factors and their effect on two measures of diffusion. The two dependent variables were internal diffusion, or spread of usage of the CPR, and infusion, or depth of usage. Data from the 144 respondents indicate that for diffusion, the organizational variables of "decision making" and "planning" had a significant impact, although the relation between "planning" and diffusion was negative. For infusion, the Innovation Attributes variable "visibility" was significant. The major implication is that successfully encouraging usage of the CPR entails attention and resources devoted to managing the organizational aspects of implementation. PMID- 9357713 TI - Improving information management in family practice: testing an adult learning model. AB - Information management training has been neglected in family practice in the UK in the past. An adult learning model for such training is introduced. A pilot study using the adult learning approach showed improvements in information management processes over the six-month study period. The research project described in this paper compares the effectiveness of on-site training using adult learning methods, written information, and no intervention, in 33 family practices in the UK. Nine of the eleven practices in the on-site training group completed the training sessions and eight provided full data, whereas only one of the eleven practices in the written information group, and only one of the eleven practices in the control group provided full data. Preliminary analysis demonstrates that on-site training practices made considerable changes to the information systems in their practices, and appreciated the importance of high quality data, both for patient care and reporting requirements. Full comparisons of data quality and information management methods are presented, and an assessment of priority training needs for maximum benefit is made. PMID- 9357714 TI - E-mail access to NetCME: implementation of server push paradigm. AB - We describe the implementation of a Continuing Medical Education project which utilizes e-mail delivery of HTML documents to facilitate participant access to case material. HTML e-mail is displayed directly within the e-mail reader of the Netscape browser. This system of proactive educational content delivery ensures simultaneous distribution to all participants. Although a more effective method of content distribution, the system preserves user confidentiality and maintains security. HTML e-mail is non-proprietary and could be integrated into existing Internet-based educational projects to facilitate user access. PMID- 9357715 TI - Performance support concepts for Web-based informatics instruction. AB - Duke University first offered World Wide Web (WWW) based courses in Nursing Informatics in January of 1997. The first class enrolled 18 nurses who were completing either a Post-Master's Certificate Program or were near completion of their Master's degree. Courses were designed around principles of advanced nursing practice, performance support, mastery learning, and virtual learning communities. Extensive learning assessment included traditional papers, real world application projects, and a variety of pre and post-test measurements. PMID- 9357716 TI - Implementing cognitive learning strategies in computer-based educational technology: a proposed system. AB - Switching the development focus of computer-based instruction from the concerns of delivery technology to the fundamentals of instructional methodology, is a notion that has received increased attention among educational theorists and instructional designers over the last several years. Building upon this precept, a proposed methodology and computer support system is presented for distilling educational objectives into concept maps using strategies derived from cognitive theory. Our system design allows for a flexible and extensible architecture in which an educator can create instructional modules that encapsulate their teaching strategies, and mimics the adaptive behavior used by experienced instructors in teaching complex educational objectives. PMID- 9357717 TI - The development and evaluation of an adaptable computer aided instruction(CAI) program for acquiring problem solving skills in biochemistry on the WWW: The "BioChem Thinker". AB - BioChem Thinker is a CAI program that was developed to enhance problem solving skills and ability to integrate knowledge in biochemistry for medical and dental students. The program runs on a WWW browser. BioChem Thinker is adaptable, it enables the teacher to create a new problem solving assignment, or edit existing assignments without in-depth knowledge of computer programming. This provides teachers with greater independence and flexibility so as to be able to adapt the program to their own course requirements. The program was implemented and evaluated in the 3rd year biochemistry course of The Hebrew University-Hadassah Medical School. The tool used to develop Biochem Thinker can be utilized to develop similar CAI in other biomedical areas. PMID- 9357718 TI - Probabilistic predictions of penetrating injury to anatomic structures. AB - This paper presents an interactive 3D graphical system which allows the user to visualize different bullet path hypotheses and stab wound paths and computes the probability that an anatomical structure associated with a given penetration path is injured. Probabilities can help to identify those anatomical structures which have potentially critical damage from penetrating trauma and differentiate these from structures that are not seriously injured. PMID- 9357720 TI - Word frequency analysis of dictated clinical data: a user-centered approach to the design of a structured data entry interface. AB - The design of a functional interface for direct entry of physical exam data by physicians remains a formidable challenge for developers of clinical information systems. Many developers use a theoretical approach, basing the interface on a model of the structure of the information and of the user-system interaction that is developed with one or more clinical domain expert(s). We explored the use of empirical analysis as a basis for the design of a structured data entry (SDE) interface. A collection of physical examination data from actual trauma patients, dictated by trauma surgeons, was used for the analysis. Using simple parsers written in Visual BASIC, we used word frequency analysis (WFA) and manual editing to identify the frequencies of unique terms used by physicians in recording 688 HEENT and 712 LUNG physical exams. A second-pass WFA was used to determine associated descriptive terms. A simple SDE interface was created based on the results of these analyses. The interface was then evaluated by assessing the extent to which the HEENT and LUNG segments of similar physical exams could be fully recorded using the empirically-based SDE interface. Using this interface, 68% of 200 trial HEENT exams, and 85% of 200 trial LUNG exams could be fully recorded. The interface was also considered helpful in recording substantial portions of the remainder of the exams. We believe that WFA can be a useful tool for finding empirical basis for SDE design. PMID- 9357719 TI - Controlled vocabulary and design of laboratory results displays. AB - Traditional data-review displays are driven by the ancillary systems that produced the data. A different paradigm is being used at Columbia-Presbyterian Medical Center (CPMC) where a controlled medical vocabulary-the Medical Entities Dictionary (MED) is the driving force behind laboratory data-review displays. Using hierarchical and semantic networks the authors have constructed a Web-based tool that considerably simplifies the MED-editing task required to create new displays. The tool uses knowledge in the MED to extract contextually relevant hierarchic and semantic sub-nets from the MED. The tool has a sensitivity of 92.2% and a relevance of 94.7% for retrieval of terms from the MED. Based on these results and given sufficient domains' structure within controlled vocabularies, we conclude that similar algorithms will enable applications to design and generate customized displays on-the-fly. PMID- 9357721 TI - Lightweight, mobile E-mail for intra-clinic communication. AB - We have developed a mobile messaging system designed for use in the clinic setting. The system is designed to facilitate quick, informal, interactions that occur in a clinical setting, e.g., requests for assistance or information. The system includes safeguards to make sure that the sender of a message is aware if a message is not read in a timely fashion. Evaluation of the system shows message delivery was about 50% slower than our target of 30 seconds. Although the mobile device used is fairly small when combined with a radio unit, it is too bulky and users did not necessarily carry the system with them. This led to delays (over eleven minutes on average) before messages were seen. We expect that improvements in hardware and clinical software will lead to more common use of such adjunct software systems. PMID- 9357722 TI - Integrating communicative goals for real-time clinical decision support. AB - A critiquing system evaluating a physician's management plan may produce a set of individual comments that, taken together, appear repetitious or incoherent. This paper presents TraumaGEN, a system for integrating sets of possibly inter-related communicative goals into one or more coherent messages. TraumaGEN takes account of the purpose of the messages, the situation in which the messages will be received, and the social role of the system. Preliminary evaluation of TraumaGEN indicates that it produce coherent integrated messages. PMID- 9357723 TI - Acceptance of a speech interface for biomedical data collection. AB - Speech interfaces have the potential to address the data entry bottleneck of many applications is the field of medical informatics. An experimental study evaluated the effect of perceptual structure on a multimodal speech interface for the collection of histopathology data. A perceptually structured multimodal interface, using speech and direct manipulation, was shown to increase speed and accuracy. Factors influencing user acceptance are also discussed. PMID- 9357724 TI - Iterative usability testing: ensuring a usable clinical workstation. AB - Once the users' needs are determined, how does one ensure that the resulting software meets the users' needs? This paper describes our application of a process, usability testing, that is used to measure the usability of systems as well as guide modifications to address usability problems. Usability testing is not a method to elicit opinions about software, but rather a method to determine scientifically a product's level of usability. Our application of usability testing is designed to determine the current usability level of a workstation designed for the clinician's use, determine specific problems with the Clinical Workstation's usability, and then evaluate the effectiveness of changes that address those problems. PMID- 9357725 TI - The availability of unavailable information. AB - Currently, developers of decision-support systems try to integrate these systems with the electronic medical record. The drawback is a limited amount of recorded medical data. System developers who face the choice between designing an integrated 'non-inquisitive' system and an integrated 'inquisitive' system need insight into the availability of information that is being missed by the support system. Therefore, we have investigated in a simulation study, the reasons why information that was being missed from the electronic medical records of patients with asthma/COPD by reviewers, had not been recorded by general practitioners. Important reasons were: the physicians had not recorded the information explicitly, they assumed the requested information to be common knowledge, and the information was available elsewhere in the electronic medical record. Also, we investigated the reasons why information that was being missed, could not be made available by the physicians. Important reasons were: the decision had been made by another decision maker, or the physician had not recorded the information at the time of the encounter. In addition to insight into the availability of missing information, system developers need to have insight into the significance of this information for the quality of the decision support, before the final choice between a non-inquisitive and an inquisitive design can be made. PMID- 9357726 TI - Design considerations in migrating an obstetrics clinical record to the Web. AB - Recently the American College of Obstetricians and Gynecologists (ACOG) embarked on an effort to promote the development of nationally networked obstetrical records. The Laboratory of Computer Science (LCS) is collaborating with them to help achieve this goal through the development of a web-based prototype of an electronic medical record (EMR) which would allow the entry and display of typical clinical information for the obstetric patient. The process of porting a stand alone application to the web environment necessitated the development of a robust software scheme that could exploit the strengths of Web-based technologies and avoid some of the drawbacks inherent in a stateless environment. PMID- 9357727 TI - Experiences with ARTEMIS--an Internet-based telemedicine system. AB - ARTEMIS is one of the first systems to exploit the Internet/Intranet technologies for exchanging patient information among health care providers. The primary project goal was to develop and demonstrate a regional telehealth environment specifically to support real-time consultations among health care providers via a computer network, provide secure access to multi-media patient records and discharge summaries, facilitate authentication/digital sign-off, multi-media mail based referrals, and network-based dictation/transcription. A prototype is deployed in southern West Virginia in a Community Care Network (CCN). The CCN consists of providers, hospitals, clinics, laboratories, that make up one "Virtual" clinic on the "Intranet". ARTEMIS employs new technologies such as Java and JavaScript for the browser, and CORBA-based "middleware" for interoperability at the server-end. Several experiments were designed for evaluating the impact of ARTEMIS on patient care. In this paper we discuss the challenges we faced and the means by which we plan to meet these challenges. We conclude by outlining new thrust areas in which we are concentrating in our next phase of development of ARTEMIS. PMID- 9357728 TI - Implementing NCEP guidelines in a Web-based disease-management system. AB - DMS is a Web-based disease-management system, which facilitates easy access for users and close connection to hospital information systems, based on clinical practice guidelines. Currently we are prototyping DMS in the area of hyperlipidemia management. However our approach is general. For each office visit, DMS generates an encounter form with recommendations based on the National Cholesterol Education Program (NCEP) guidelines. In between visits, DMS provides email notifications to clinicians about delinquent laboratory studies and recommendations for patient management based on recently available information. By reviewing previous efforts for implementing NCEP guidelines and some of the pitfalls that were encountered, we first constructed DMS for hyperlipidemia management. A detailed description of DMS is provided in this paper. PMID- 9357729 TI - SecondOpinion: interactive Web-based access to a decision model. AB - In this paper, we describe a computer architecture, which we call SecondOpinion, designed for automated, normative patient decision support over the World Wide Web. SecondOpinion custom tailors the discussion of therapy options for patients by eliciting their preferences for relevant health states via an interactive WWW interface and then integrating those results in a decision model. The SecondOpinion architecture uses a Finite State Machine representation to track the course of a patient's consultation and to choose the next action to take. The consultation has five distinct types of interactions: explanation of health states, assessment of preferences, detection and correction of errors in preference elicitations, and feedback on the implications of preference. A linear "summary model" speeds calculations of predictions from the decision model and makes it possible to dynamically calculate 95% confidence intervals for the marginal utility of each treatment option. Preferences for states are assessed in the order of their variance contribution to the models predictions in an iterative fashion. Only the states required to obtain a 95% Confidence Interval (CI) that excludes zero are assessed. In Monte Carlo simulation studies, the average number of utility assessments required for the 95% CI to exclude zero in an individual was 4.24 (SD = 1.97) out of 8 relevant health states. the SecondOpinion architecture provides an efficient, "discussion-like" experience leading to an individual-specific treatment recommendation. It may be a cost effective approach to bring decision analytic advice to the bedside. PMID- 9357730 TI - SAM: speech-aware applications in medicine to support structured data entry. AB - In the last two years, improvement in speech recognition technology has directed the medical community's interest to porting and using such innovations in clinical systems. The acceptance of speech recognition systems in clinical domains increases with recognition speed, large medical vocabulary, high accuracy, continuous speech recognition, and speaker independence. Although some commercial speech engines approach these requirements, the greatest benefit can be achieved in adapting a speech recognizer to a specific medical application. The goals of our work are first, to develop a speech-aware core component which is able to establish connections to speech recognition engines of different vendors. This is realized in SAM. Second, with applications based on SAM we want to support the physician in his/her routine clinical care activities. Within the STAMP project (STAndardized Multimedia report generator in Pathology), we extend SAM by combining a structured data entry approach with speech recognition technology. Another speech-aware application in the field of Diabetes care is connected to a terminology server. The server delivers a controlled vocabulary which can be used for speech recognition. PMID- 9357731 TI - Can data representation and interface demands be reconciled? Approach in ORCA. AB - Research in the domain of computer-based patient records had always faced the conflicting demands of efficiency for the practicing physician and suitability of the record contents for data analysis in view of decision support, research, and quality assessment. Interface and contents pose different demands on the data model underlying the record. The challenge is to combine the most suitable model for data representation with the interface that best fits the clinical setting. ORCA (Open Record for CAre) provides a solution by making the distinction between domain dependent and domain independent data and letting domain dependence be decisive for the choice of model. Interactive definition of custom-views provides interface flexibility for domain dependent data. Views on domain independent data need not cope with the limitations of multiple table views in relational DBMSs. A standard set of single table queries can support recording of domain independent data, irrespective of the clinical setting. PMID- 9357732 TI - Preserving context in a multi-tasking clinical environment: a pilot implementation. AB - The Partners Clinical Application Suite (CAS) is a multi-tasking software architecture that facilitates the development, deployment, and use of advanced clinical information management applications. This paper describes 1) a software shell in which clinical applications run; 2) an application programming interface (API); and 3) development of a set of "Look & Feel" guidelines. Through its emphasis on support for multi-tasking and application interoperability, CAS facilitates preservation of the user's context. PMID- 9357733 TI - Data collection and information presentation for optimal decision making by clinical managers--the Autocontrol Project. AB - The Autocontrol methodology has been developed in order to support the optimisation of decision-making and the use of resources in the context of a clinical unit. The theoretical basis relates to quality assurance and information systems and is influenced by management and cognitive research in the health domain. The methodology uses population rather than individual decision making and because of its dynamic feedback design promises to have rapid and profound effect on practice. Most importantly the health care professional is the principle user of the Autocontrol system. In this methodology we distinguish three types of evidence necessary for practice change: practice based or internal evidence, best evidence derived from the literature or external evidence concerning the practice in question, and process based evidence on how to optimise the process of practice change. The software used by the system is of the executive decision support type which facilitates interrogation of large databases. The Autocontrol system is designed to interrogate the data of the patient medical record however the latter often lacks data on concomitant resource use and this must be supplemented. This paper reviews the Autocontrol methodology and gives examples from current studies. PMID- 9357734 TI - Web-based data integration and annotation in the intensive care unit. AB - Integrating patient data to facilitate review and annotation is a challenging task in the intensive care unit (ICU), partially due to the wide variety of proprietary systems and types of data involved. This paper describes SIMON-Web, a Java-based user interface to integrate data from an existing bedside monitoring system and a clinical information system (CIS). SIMON-Web displays graphical data from physiologic monitors and other bedside devices as well as information from the clinical laboratory and physician order-entry systems, and allows users to annotate the data using a point-and-click or free-text interface. By continuing to add functionality to SIMON-Web's extensible user interface, we hope to continue to augment and eventually replace manual care provider data charting in the ICU. As of March, 1997, SIMON-Web has been implemented on two bedside personal computer workstations in the cardiac care unit (CCU) at Vanderbilt University Medical Center. PMID- 9357735 TI - Web client and ODBC access to legacy database information: a low cost approach. AB - A new method has been developed for the Department of Orthopaedics of Vanderbilt University Medical Center to access departmental clinical data. Previously this data was stored only in the medical center's mainframe DB2 database, it is now additionally stored in a departmental SQL database. Access to this data is available via any ODBC compliant front-end or a web client. With a small budget and no full time staff, we were able to give our department on-line access to many years worth of patient data that was previously inaccessible. PMID- 9357736 TI - Electronic forms: benefits drawbacks of a World Wide Web-based approach to data entry. AB - It has long been realized that, compared to paper-based records, electronic record systems provide many advantages in the healthcare environment, including increased availability, improved legibility, long-term accessibility, (potentially) greater completeness, data encoding, and automated decision support and analysis. In spite of these recognized benefits, collection of patient data at the point of service generally does not occur, in large part because each such effort usually requires application-specific software and hardware, and, most significantly, provider time. Given the presence of WWW browsers now available on nearly every desktop, the support and access concerns for data entry applications can be substantially lessened. Despite these advantages, there are also downsides to the use of the WWW for data entry, including user interface issues and security. At CPMC, we are currently using web-based forms to gather patient charge data from physical and occupational therapists. Benefits of this approach have included a 98.2% user compliance rate for at least weekly data entry, and the reduction of charge posting from an average of 24.3 days to 2.3 days following the date of service. Drawbacks to WWW-based applications have included increased security exposure and persistent human tendencies to enter data in batches rather than at the time of service. A final conclusion was that, in the absence of a strong central mandate, providers must perceive a clear benefit in order to be willing to learn and use a new technology. PMID- 9357737 TI - Meeting clinician information needs by integrating access to the medical record and knowledge resources via the Web. AB - MINDscape is a web based integrated interface to diverse sources of clinical information including both patient specific information (electronic medical record) as well as medical knowledge (the "digital library") to provide "just in time" information at the point of care. It was developed at the University of Washington to meet clinical information needs both as identified locally and by a review of the literature. Beta testing by over 600 clinicians is in progress and medical centers wide access scheduled for Fall 1997. We describe the information needs we sought to meet and the ongoing evaluation approach we are taking to ensure the information needs of a diverse group of clinicians are met. The iterative evolution of the interface from prototype, to alpha to large scale beta testing is reported. Integration of information occurs at three levels: integration of information by patient, integration of information by provider, and integration of patient specific information with medical reference material and decision support tools. PMID- 9357738 TI - A natural language parsing system for encoding admitting diagnoses. AB - Free-text or natural language documents make up an increasing part of the computerized medical record. While they do provide accessible clinical information to health care personnel, they fail to support processes that require clinical data coded according to a shared lexicon and data structure. We have developed a natural language parser that converts free-text admitting diagnoses into a coded form. This application has proven acceptably accurate in the experimental laboratory to warrant a test in the target clinical environment. Here we describe an approach to moving this research application into a production environment where it can contribute to the efforts of the Health Information Services Department. This transition is essential if the products of natural language understanding research are to contribute to patient care in a routine and sustainable way. PMID- 9357739 TI - Text structures in medical text processing: empirical evidence and a text understanding prototype. AB - We consider the role of textual structures in medical texts. In particular, we examine the impact the lacking recognition of text phenomena has on the validity of medical knowledge bases fed by a natural language understanding front-end. First, we review the results from an empirical study on a sample of medical texts considering, in various forms of local coherence phenomena (anaphora and textual ellipses). We then discuss the representation bias emerging in the text knowledge base that is likely to occur when these phenomena are not dealt with--mainly the emergence of referentially incoherent and invalid representations. We then turn to a medical text understanding system designed to account for local text coherence. PMID- 9357740 TI - Searching for information on the Internet using the UMLS and Medical World Search. AB - Medical World Search is a search engine for medical information on the Internet that distinguishes itself from other search engines by its built-in knowledge of medical terminology through its use of the National Library of Medicine's UMLS and its carefully selected but large database of medical sites. After discussing some of the previous uses of the UMLS for medical information retrieval, we describe the Medical World Search system. In October 1996, Medical World Search became operational on the World Wide Web at http:@www.mwsearch.poly.edu. It has been operating uninterrupted since then. We review our experiences with creating a search engine for medical information on the Internet and using the UMLS in this application. The UMLS has some clear advantages in this application. Some aspects of the UMLS also decrease its usefulness in information retrieval. Medical World Search's usage by medical information seekers is summarized. future directions for research are outlined. PMID- 9357741 TI - Identification of findings suspicious for breast cancer based on natural language processing of mammogram reports. AB - There is need for encoded data for computerized clinical decision support, but most such data are unavailable as they are in free-text reports. Natural language processing offers one alternative for encoding such data. MedLEE is a natural language processing system which is in routine use for encoding chest radiograph and mammogram reports. In this paper, we study MedLEE's ability to identify mammogram findings suspicious for breast cancer by comparing MedLEE's encoding with a logbook of all suspicious findings maintained by the mammography center. While MedLEE was able to identify all the suspicious findings, it varied in the level of granularity, particularly about the location of the suspicious finding. Thus, natural language processing is a useful technique for encoding mammogram reports in order to detect suspicious findings. PMID- 9357743 TI - In vitro evaluation of a glass-ceramic restorative material. AB - The aim of the present study was to evaluate the clinically relevant properties of the recently introduced ceramic material IPS Empress, which is marketed for all-ceramic restorations. The following parameters were investigated: three- and four-point bending strength, bi-axial flexure strength, compressive and diametral tensile strength, compressive strength and marginal fit of full crowns. The results show that this material is a highly developed glass-ceramic with physical properties making this dental material well suitable for adhesively luted restorations. PMID- 9357742 TI - The closure of the gingival crevice following gingival retraction for impression making. AB - Scant attention has been paid to the effectiveness of chemomechanical displacement of the gingiva prior to impression making for fixed partial dentures. The closure of the gingival crevice following removal of medicated retraction cord was observed using a miniature video camera. Sulcular widths were measured at time intervals at the midbuccal and transitional line angle areas. The closure rate of the transitional line angle area was significantly faster than that of the mid-buccal area during the first 90 s. An average sulcular width of 0.2 mm was reached at the transitional line angle after less than 30 s. PMID- 9357744 TI - Marginal adaptation of a sintered ceramic inlay system before and after cementation. AB - The long term clinical performance of porcelain inlays depends on a number of factors and the marginal adaptation is one of significant interest. The purpose of this in vitro study was to evaluate the marginal integrity of a sintered inlay technique (Ducera), before and after cementation. MOD cavities without bevels were prepared on 10 human mandibular molar teeth and porcelain inlays were fabricated according to the manufacturer's instructions. Inlays were evaluated microscopically for their adaptation to the occlusal and approximal margins of the tooth by means of a replica technique. Inlays were cemented with a dual-cured hybrid composite luting material (Enforce). After polishing, each tooth was sectioned in buccal/lingual and mesial/distal directions following the same procedure in the sectioning of replicas. The marginal gap and the thickness of exposed cement were measured at each section. The mean marginal gap of 71.83 +/- 8.93 microm recorded for the occlusal margin before cementation was significantly smaller than that of 105.6 +/- 39.33 microm calculated at the approximal margin. Following the cementation, the adaptation of the inlays at the occlusal margin was also found to be superior to that of the approximal margin. Comparison of mean gap values before and after cementation revealed that the marginal gap increased by 6.94 microm and 23.25 microm at the occlusal and approximal margins, respectively. Although polishing was performed after cementation, excess luting material was still observed, that caused an increase in the width of the exposed luting cement. PMID- 9357745 TI - Characteristics of masticatory movement in relation to inclination of occlusal plane. AB - The relationship between masticatory movement and the inclination of the occlusal plane in sagittal plane was analysed in 41 young adults. It was found that the occlusal plane and the masticatory closing path were consistent in maintaining an almost perpendicular relationship with each other, regardless of the variation in inclination of the occlusal plane. This finding can be explained by the observation that the timing of the balancing-side condylar translation during closure correlated with the inclination of the occlusal plane. The inclination of the occlusal plane also influenced the masticatory closing pattern in the sagittal plane. Anterior convex closure patterns dominated when the occlusal plane inclined in the anterior direction. In contrast, the majority of posterior convex closure patterns were induced by the posteriorly inclined occlusal plane. The appearance of these types seems to reflect a harmonious relationship between the inclination of the occlusal plane, tooth guidance, and other central and peripheral control. The correlation between the inclination of the occlusal plane and masticatory closing movement could serve as the functional background for the significance of the occlusal plane. PMID- 9357746 TI - A long-term study of transverse stability of maxillary teeth in patients with unilateral complete cleft lip and palate. AB - The aim of this study was to investigate the long term post-treatment transverse stability of the maxillary dental arch in subjects with unilateral complete cleft lip and palate (UCLP) treated by the Harvold/Bohn method of orthodontic expansion and prosthodontic retention. The treatment of 22 consecutive patients, primarily operated on during the period 1957-60, was completed at a mean age of 18.1 years by the provision of a fixed partial retention prosthesis across the cleft using the cleft side central incisor and canine only as abutment teeth. The cleft side lateral incisor was missing in each case. Dental casts were made at the time of abutment preparation and at six subsequent times with the final observation 13.5 years after treatment completion. Measurements of any shift in the transverse position of cleft side and non-cleft side canines, premolars and first molars were made on standardized photographs of the casts. A constructed anteroposterior palatal line served as 'midline' reference. A mean reduction of width at the final observation, as recorded from the palatal surface to the reference line, was for the cleft side canine: -0.4 mm, the premolar immediately distal to the prosthesis and the first molar: both -1.2 mm. The corresponding mean width reductions on the noncleft side were: canine -0.9 mm, premolar -1.2 mm, first molar -1.6 mm. The rate of movement towards the midline decreased linearly with In(time) for all variables (P < 0.02) but for the cleft side canine. PMID- 9357748 TI - An optical system for measuring inclination and area of occlusal facets. AB - Occlusal facets are regarded as an individual record of occlusal contacts that result from various mandibular movements. Analysing characteristics of the facets is important for the functional assessment of the stomatognathic system. A new system which quantifies such characteristics optically has been developed. This system is composed of optical devices, a graphic image-processor, a set of stages for mounting and positioning the model and laser indicators of the measuring point. The system enables the inclination and area of each facet and its location to be measured and its processed image displayed on the graphic monitor. PMID- 9357750 TI - Study of jaw movement and masticatory muscle activity during unilateral chewing with and without balancing side molar contacts. AB - This research evaluated the relationship between balancing side molar contacts and chewing patterns measured with a jaw movement analyser and multi-channel electromyography (EMG) of the masticatory muscles. Nine healthy subjects with relatively normal occlusions participated in the experiment and were divided into those with balancing side molar contacts and those without. The block gum chewing task was performed on each side of the mouth for 10 s. The results showed more asymmetrical levels of jaw closing muscle activity during unilateral chewing in the group with balancing side molar contacts when compared with the group without these contacts. PMID- 9357749 TI - The effect of posterior tooth guidance on non-working side arbitrary condylar point movement. AB - Occlusal form is frequently modified in clinical practice and yet we do not have detailed knowledge of the possible effects of these changes on condylar movement. The aim of this study was to quantify the effects of an alteration in the occlusion on condylar movement during a lateral excursive jaw movement. Posterior tooth guidances (i.e. metal overlays) were attached to both maxillary first molars. The movement of arbitrary condylar points on the non-working side was recorded in seven subjects during lateral excursion under natural tooth guidance (control) and was compared with that after placement of the overlays (guidance). The guidance resulted in statistically significant changes to the displacement of the arbitrary condylar points on the non-working side. For example, at a standardized (3 mm) displacement along the mid-incisor point trajectory during the lateral excursion for both control and guidance in all subjects, the corresponding displacements of the condylar points were statistically significantly decreased under the guidance situation in comparison with the control situation. These data suggest that, for the same magnitude of mandibular displacement during lateral excursion, the introduction of a posterior tooth guidance limits condylar displacement on the non-working side. PMID- 9357747 TI - Clinical evaluation of cervical dentine sensitivity in a population of patients referred to a specialist periodontology department: a pilot study. AB - The prevalence of tooth sensitivity [Cervical Dentine Sensitivity (CDS)] in adult populations indicates that 8-35% of subjects reported CDS depending on the population studied and the methodology used. Few studies, however, have reported on the prevalence of CDS in periodontal patients. The aim of the study was to determine the prevalence, severity and distribution of CDS in patients referred for specialist periodontal diagnosis. Fifty-one patients [27 male, 24 female; mean age 48.5 years (standard deviation 11.63)] who gave their informed written consent were clinically evaluated for CDS using recognized methods of assessment, namely Yeaple probe, cold air blast and subjective evaluation. Other clinical variables (e.g. plaque and recession scores) were also recorded at this visit. Regression analysis and correlation coefficients were used to determine the relationship between the clinical variables. The results demonstrated a prevalence of CDS ranging between 72.5 and 98% of patients, with no significant gender difference. Results for the distribution of tooth types showed that molar teeth were mainly affected, followed by left canines and premolars. No correlation was noted between plaque, recession, response to tactile or thermal stimulation. Pain response from tactile and thermal stimulation showed no significant difference between tooth surfaces. Cold stimulation was perceived to be the dominant pain-producing stimulus as had been previously reported. The results of this investigation support those from another study, which found the prevalence of CDS to be higher in periodontal patients than has been reported elsewhere. This finding would suggest that previous periodontal treatment and/or periodontal disease may play a role in the aetiology of CDS. PMID- 9357751 TI - Posterior bridges retained by resin-bonded cast metal inlay retainers: a report of 60 cases followed for 6 years. AB - Between May 1989 and July 1994, 60 adhesive fixed partial posterior dentures retained by resin-bonded cast metal inlays were placed under controlled conditions. The influence of various prognostic factors on the event free and overall service duration was investigated with univariate and multivariate analysis. The majority of the failures (16 of 18) were caused by loss of adhesion at the metal cement interface and were observed either as debonding of the restorations completely or dislodgement of a single retainer. The other two failures appeared as secondary caries. Univariate analysis demonstrated that, retainer type, approximal configuration and dentine exposition had no effect on the event free service duration. On the other hand, gingival finishing level and luting agent were found to have an effect. Univariate testing was also conducted for the overall service duration and none of the variables were found to have an effect. Multivariate Cox Regression Analysis was performed to estimate the influence of categorical covariates: type of retainer, approximal preparation modification, gingival finishing level, preparation depth and luting agent on survival rates of event free and overall service duration. Luting agent was found to be the single independent prognostic variate (P < 0.0001) for the event free service duration and the other covariates were rejected. For overall service duration, none of the variables were found to be effective. PMID- 9357752 TI - Autoradiographic determination of marginal leakage of a pressed glass ceramic inlay. AB - The marginal integrity and microleakage of pressed glass ceramic inlays were evaluated using autoradiography. IPS/Empress ceramic inlays were fabricated for 10 human molar mandibular teeth. After adjusting the inlays, they were etched with 37% phosphoric acid gel for 30 s and silanized with Monobond S for 30 s. Before cementation with dual cure resin cement the inlays and cavity walls were gently covered with a thin layer of bonding agent. When the cementation process was completed the samples were cycled 300 times between a 55 degrees C hot bath and a 5 degrees C cold bath. The samples were placed in each bath for 60 s, with 5 s intervals between immersions, then the specimens were immersed in an aqueous solution of Ca-45. After 24 h the inlay and tooth assemblies were removed, rinsed with water and placed in cold-cured acrylic resin, then sectioned through the long axis for autoradiographic analysis. According to the penetration of Ca-45, the microleakage level was scored for each section. The results indicated slight penetration of Ca-45 on autoradiographic films. PMID- 9357753 TI - Three-dimensional cell cultures: from molecular mechanisms to clinical applications. AB - This article reviews actual advances in the development and application of three dimensional (3-D) cell culture systems. Recent therapeutically oriented studies include characterization of multicellular-mediated drug resistance, novel ways of quantifying hypoxia, and new approaches to more efficient immunotherapy. Recent progress toward understanding the development of necrosis in tumor spheroids has been made using novel spheroid models. 3-D cultures have been used for studies on molecular mechanisms involved in invasion and metastasis, with a major focus on the role of E-cadherin. Similarly, tumor angiogenesis and the significance of vascular endothelial growth factor have been investigated in a variety of 3-D culture systems. There are many ongoing developments in tissue modeling or remodeling that promise significant progress toward the development of bioartificial liver support and artificial blood. Perhaps one of the most interesting areas of basic research with 3-D cultures is the characterization of embryoid bodies obtained from stable embryonic stem cells. These models have greatly increased the understanding of embryonic development, in particular through the notable exceptional advances in cardiogenesis. PMID- 9357754 TI - Glutamine utilization by rat neutrophils: presence of phosphate-dependent glutaminase. AB - The capacity of rat neutrophils to utilize glutamine was investigated by 1) determination of oxygen consumption in the presence of glucose or glutamine, 2) measurement of maximal activity of phosphate-dependent glutaminase, 3) Northern blot, Western blot, and immunocytochemical detection of glutaminase, and 4) measurement of glutamine utilization and also production of ammonia, glutamate, aspartate, alanine, and lactate and decarboxylation of [U-14C]glutamine in cells incubated for 1 h. The rate of respiration by isolated neutrophils in the absence of added substrate was 5.0 nmol x min(-1) x 10(7) cells(-1). Maximal activity of phosphate-dependent glutaminase was 56 nmol x min(-1) x mg protein(-1) in freshly obtained neutrophils; the Michaelis-Menten constant was 3.5 mM for glutamine. This enzyme activity was inhibited by 2 mM glutamate, 2 mM oxoglutarate, and 2 mM NH4Cl. The presence of glutaminase protein (65 kDa) was confirmed by Western blot and immunocytochemical detection and the presence of the mRNA (6.0 kb) by Northern blot analysis. Glutamine was utilized by neutrophils incubated for 1 h at a rate of 12.8 nmol x min(-1) x mg protein(-1) when the amino acid was added to the medium at 2 mM, which is three to four times higher than the physiological concentration. In the presence of 0.5 mM glutamine, the amino acid was utilized at a rate of 2.9 nmol x min(-1) x mg protein(-1). The addition of 0.5 mM glutamate to the incubation medium caused a marked reduction (by 70%) in glutamine utilization by neutrophils. Glucose was utilized at 7.7 nmol x min(-1) x mg protein(-1) when cells were incubated in 5 mM glucose. The conversion of [U 14C]glutamine to 14CO2 was very low: <1% was totally oxidized. The formation of ammonia was approximately 27% of glutamine utilization, and the conversion of glutamine to glutamate, aspartate, alanine, and lactate accounted for approximately 84.6% of the total amino acid utilized by neutrophils. In this study, evidence is presented that, in addition to lymphocytes and macrophages, neutrophils also utilize glutamine. PMID- 9357755 TI - Intestinal ischemia and reperfusion injury in transgenic mice overexpressing copper-zinc superoxide dismutase. AB - Superoxide dismutase (SOD) scavenges oxygen radicals that are implicated in the pathogenesis of intestinal ischemia-reperfusion injury. The effect of intestinal ischemia and reperfusion was investigated in transgenic mice overexpressing human Cu-Zn SOD. Ischemia was induced by occluding the superior mesenteric artery. Myeloperoxidase activity was determined as an index of neutrophil infiltration, and malondialdehyde levels were measured as an indicator of lipid peroxidation. Forty-five minutes of intestinal ischemia followed by 4 h of reperfusion caused an increase in intestinal levels of malondialdehyde in both nontransgenic and transgenic mice, but the concentration of malondialdehyde was significantly greater in nontransgenic mice. Intestinal ischemia-reperfusion also caused an increase in intestinal and pulmonary myeloperoxidase activity in nontransgenic and transgenic mice, but the transgenic mice had significantly lower levels of myeloperoxidase activity than nontransgenic mice. Transgenic mice had higher levels of intestinal SOD activity than nontransgenic mice. There were no significant differences in the catalase or glutathione peroxidase activities. In conclusion, our study demonstrates that the overexpression of SOD protects tissues from neutrophil infiltration and lipid peroxidation during intestinal ischemia-reperfusion. PMID- 9357756 TI - Role of beta1- and beta3-adrenoceptors in the regulation of lipolysis and thermogenesis in rat brown adipocytes. AB - To evaluate the physiological functions of beta1-, beta2-, and beta3 adrenoceptors (ARs) in brown adipose tissue, the lipolytic and respiratory effects of various adrenergic agonists and antagonists were studied in rat brown adipocytes. The beta-agonists stimulated both lipolysis and respiration (8-10 times above basal levels), with the following order of potency (concentration eliciting 50% of maximum response): CL-316243 (beta3) > BRL-37344 (beta3) > isoproterenol (mainly beta1/beta2) > norepinephrine (NE; mainly beta1/beta2) > epinephrine (mainly beta1/beta2) >> dobutamine (beta1) >> procaterol (beta2). Schild plot coefficients of competitive inhibition experiments using ICI-89406 (beta1 antagonist) revealed that more than one type of receptor mediates NE action. It is concluded from our results that 1) NE, at low plasma levels (1-25 nM), stimulates lipolysis and respiration mainly through beta1-ARs, 2) NE, at higher levels, stimulates lipolysis and respiration via both beta1- and beta3 ARs, 3) beta2-ARs play only a minor role, and 4) beta3-ARs may represent the physiological receptors for the high NE concentrations in the synaptic cleft, where the high-affinity beta1-ARs are presumably desensitized. It is also suggested that lipolysis represents the flux-generating step regulating mitochondrial respiration. PMID- 9357758 TI - Hyaluronic acid-specific regulation of cytokines by human uterine fibroblasts. AB - The physiological inflammatory response can provide an effective mechanism for delivering the baby at the time of parturition. We characterized the mechanisms by which hyaluronic acid (HA) regulates interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and interleukin-8 (IL-8) production in human uterine fibroblasts. A dose-dependent increase in cytokine release was observed over an HA concentration range of 10 microg/ml to 1 mg/ml. The action of HA on the cytokine production is mediated by CD44. Under serum-free conditions, HA induced cytokine generation was significantly less compared with production in the presence of serum, suggesting involvement of serum proteins. Addition of inter-alpha-trypsin inhibitor (ITI) under serum-free conditions enhanced the HA induced synthesis of TNF-alpha, which stimulated the temporary release of IL-8. In addition, HA and IL-1beta stimulated the release of hyaluronidase by the fibroblasts. These results indicate that cytokine production in human uterine fibroblasts is regulated in a CD44-HA-ITI-specific fashion. HA may be involved in the regulation of delivery in part through the selective release of cytokines that contribute to uterine cervical ripening. PMID- 9357757 TI - Expression of dopamine D2 receptor in PC-12 cells and regulation of membrane conductances by dopamine. AB - PC-12 cells depolarize during hypoxia and release dopamine. The hypoxia-induced depolarization is due to inhibition of an O2-sensitive K+ current. The role of dopamine released during hypoxia is uncertain, but it could act as an autocrine to modulate membrane conductance during hypoxia. The current study was undertaken to investigate this possibility. Reverse transcription-polymerase chain reaction and sequence analysis revealed that the D2 isoform of the dopamine receptor is expressed in rat PC-12 cells. Exogenously applied dopamine and the D2 agonist quinpirole elicited inhibition of a voltage-dependent K+ current (I(K)) that was prevented by sulpiride, a D2 receptor antagonist. Dopamine and quinpirole applied during hypoxia potentiated the inhibitory effect of hypoxia on I(K). We also found that quinpirole caused reversible inhibition of a voltage-dependent Ca2+ current (I(Ca)) and attenuation of the increase in intracellular free Ca2+ during hypoxia. Our results indicate that dopamine released from PC-12 cells during hypoxia acts via a D2 receptor to "autoregulate" I(K) and I(Ca). PMID- 9357759 TI - Activation of NF-kappaB in intestinal epithelial cells by enteropathogenic Escherichia coli. AB - The initial response to infection is recruitment of acute inflammatory cells to the involved site. Interleukin (IL)-8 is the prototypical effector molecule for this process. Transcription of the IL-8 gene is primarily governed by the nuclear transcription factor (NF)-kappaB. Intestinal epithelial cells produce IL-8 in response to infection by enteric pathogens yet remain quiescent in a milieu where they are literally bathed in normal bacterial flora. We therefore sought to investigate NF-kappaB activation in response to enteropathogenic Escherichia coli (EPEC), nonpathogenic E. coli, and bacterial lipopolysaccharide in an intestinal epithelial cell (T84) model and to determine whether EPEC-induced activation of NF-kappaB factor is causally linked to IL-8 production. We report herein that NF kappaB is activated by EPEC, yet such a response is not extended to nonpathogenic organisms or purified E. coli lipopolysaccharide. Transcription factor decoys significantly diminished IL-8 production in response to EPEC, demonstrating a causal relationship. Furthermore, deletion of specific EPEC virulence genes abrogates the NF-kappaB-activating property of this pathogen, suggesting that specific bacterial factors are crucial for inducing this response. These studies show for the first time that infection of intestinal epithelial cells with EPEC activates NF-kappaB, which in turn initiates IL-8 transcription, and highlight the differential response of these cells to bacterial pathogens vs. nonpathogens. PMID- 9357760 TI - Ca2+-sensing receptors in intestinal epithelium. AB - Expression of Ca2+-sensing receptors (CaR) was demonstrated in several human intestinal epithelial cell lines (T84, HT-29, and Caco-2) and in rat intestinal epithelium by both reverse transcriptase-polymerase chain reaction (PCR) and Northern blotting of RNA. Restriction patterns of the PCR products were of the sizes predicted by the human and rat sequences. CaR agonists (Ca2+, poly-L arginine, protamine) mediated an increase in intracellular Ca2+ in HT-29-18-C1 cells (monitored by changes in fura 2 fluorescence), which was dependent on release from thapsigargin-sensitive stores. U-73122, an inhibitor of phosphatidylinositol-phospholipase C, eliminated the CaR agonist-mediated rise in intracellular Ca2+, whereas its inactive analog, U-73343, had no effect. Pertussis toxin pretreatment had no effect on CaR agonist-mediated modulation of intracellular Ca2+. Taken together, these studies demonstrate that CaR are expressed in intestinal epithelial cells and couple to mobilization of intracellular Ca2+. The presence of CaR in intestinal epithelial cells presents a new locus for investigations into the role(s) of extracellular Ca2+ in modulating intestinal epithelial cell differentiation and transepithelial Ca2+ transport. PMID- 9357762 TI - Carbachol induces oscillations in membrane potential and intracellular calcium in a colonic tumor cell line, HT-29. AB - The patch-clamp technique was used to study the effects of carbachol (CCh) on HT 29 cells. During CCh exposure, the cells (n = 23) depolarized close to the equilibrium potential for Cl- (E(Cl-); -48 mV) and the membrane potential then started to oscillate (16/23 cells). In voltage-clamp experiments, similar oscillations in whole cell currents could be demonstrated. The whole cell conductance increased from 225 +/- 25 pS in control solution to 6,728 +/- 1,165 pS (means +/- SE, n = 17). In substitution experiments (22 mM Cl- in bath solution, E(Cl-) = 0 mV), the reversal potential changed from -41.6 +/- 2.2 mV (means +/- SE, n = 9) to -3.2 +/- 2.0 mV (means +/- SE, n = 7). When the cells were loaded with the calcium-sensitive fluorescent dye, fluo 3, and simultaneously patch clamped, CCh caused a synchronous oscillating pattern of fluorescence and membrane potential. In cell-attached patches, the CCh-activated currents reversed at a relative membrane potential of 1.9 +/- 3.7 mV (means +/- SE, n = 11) with control solution in the pipette and at 46.2 +/- 5.3 mV (means +/ SE, n = 10) with a 15 mM Cl- solution in the pipette. High K+ (144 mM) did not change the reversal potential significantly (P < or = 0.05, n = 8). In inside-out patches, calcium-dependent Cl- channels could be demonstrated with a conductance of 19 pS (n = 7). It is concluded that CCh causes oscillations in membrane potential that involve calcium-dependent Cl- channels and a K+ permeability. PMID- 9357761 TI - Cloning and functional expression of a ClC Cl- channel from the renal cell line A6. AB - Cl- channels are important for ion transport and cell volume regulation in A6 renal cells. In the present study, we used reverse transcriptase (RT)-polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) to identify proteins homologous to ClC Cl- channel proteins in A6 cells. Using degenerate primers designed on consensus sequences for members of the ClC family, we amplified an RT-PCR product that had significant homology to the ClC sequences. RACE-PCR was then used to isolate several full-length clones that had total lengths from 2,764 to 3,016 base pairs. Although the coding regions were identical, sequence differences occurred in the 5' noncoding regions. The amino acid sequences of the clones had high homologies to rat and human ClC-5 (85 and 84%, respectively, if the 5th methionine of the open reading frame represents the start codon). Three parts of the protein (53, 80, and 63 amino acids in length) were 97-100% homologous to the mammalian sequences. Ribonuclease protection assay analysis revealed mRNA for this protein in oocytes, kidney, intestine, liver, brain, and blood, with lower amounts in stomach, muscle, and skin. Expression of the clones in Xenopus laevis oocytes resulted in an outwardly rectifying Cl- current that was inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and possessed an anion selectivity of I- > Cl- >> gluconate. PMID- 9357763 TI - Functional upregulation of H+-ATPase by lethal acid stress in cultured inner medullary collecting duct cells. AB - The response of H+-ATPase to lethal acid stress is unknown. A mutant strain (called NHE2d) was derived from cultured inner medullary collecting duct cells (mIMCD-3 cells) following three cycles of lethal acid stress. Cells were grown to confluence on coverslips, loaded with 2',7'-bis(carboxyethyl)-5(6) carboxyfluorescein, and monitored for intracellular pH (pHi) recovery from an acid load. The rate of Na+-independent pHi recovery from an acid load in mutant cells was approximately fourfold higher than in parent cells (P < 0.001). The Na+ independent H+ extrusion was ATP dependent and K+ independent and was completely inhibited in the presence of diethylstilbestrol, N,N'-dicyclohexylcarbodiimide, or N-ethylmaleimide. These results indicate that the Na+-independent H+ extrusion in cultured medullary cells is mediated via H+-ATPase and is upregulated in lethal acidosis. Northern hybridization experiments demonstrated that mRNA levels for the 16- and 31-kDa subunits of H+-ATPase remained unchanged in mutant cells compared with parent cells. We propose that lethal acid stress results in increased H+-ATPase activity in inner medullary collecting duct cells. Upregulation of H+-ATPase could play a protective role against cell death in severe intracellular acidosis. PMID- 9357765 TI - F-actin modulates swelling-activated chloride current in cultured chick cardiac myocytes. AB - The integrity of F-actin and its association with the activation of a Cl- current (I(Cl)) in cultured chick cardiac myocytes subjected to hyposmotic challenge were monitored by whole cell patch clamp and fluorescence confocal microscopy. Disruption of F-actin by 25 microM cytochalasin B augmented hyposmotic cell swelling by 51% (from a relative volume of 1.54 +/- 0.10 in control to 2.33 +/- 0.21), whereas stabilization of F-actin by 20 microM phalloidin attenuated swelling by 15% (relative volume of 1.31 +/- 0.05). Trace fluorochrome-labeled (fluorescein isothiocyanate or tetramethylrhodamine isothiocyanate) phalloidin revealed an intact F-actin conformation in control cells under hyposmotic conditions despite the considerable changes in cell volume. Sarcoplasmic F-actin was very disorganized and occurred only randomly beneath the sarcolemma in cells treated with cytochalasin B, whereas no changes in F-actin distribution occurred under either isosmotic or hyposmotic conditions in cells treated with phalloidin. Swelling-activated I(Cl) (68.0 +/- 6.0 pA/pF at +60 mV) was suppressed by both cytochalasin B (22.7 +/- 5.1 pA/pF) and phalloidin (22.5 +/- 3.5 pA/pF). On the basis of these results, we suggest that swelling of cardiac myocytes initiates dynamic changes in the cytoarchitecture of F-actin, which may be involved in the volume transduction processes associated with activation of I(Cl). PMID- 9357764 TI - Effects of PKC alpha activation on Ca2+ pump and K(Ca) channel in deoxygenated sickle cells. AB - We have previously shown that a pretreatment with phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C (PKC), reduced deoxygenation-induced K+ loss and Ca2+ uptake and prevented cell dehydration in sickle anemia red blood cells (SS cells) (H. Fathallah, E. Coezy, R.-S. De Neef, M.-D. Hardy-Dessources, and F. Giraud. Blood 86: 1999-2007, 1995). The present study explores the detailed mechanism of this PMA-induced inhibition. The main findings are, first, the detection of PKC alpha and PKC zeta in normal red blood cells and the demonstration that both isoforms are expressed at higher levels in SS cells. The alpha-isoform only is translocated to the membrane and activated by PMA and by elevation of cytosolic Ca2+. Second, PMA is demonstrated to activate Ca2+ efflux in deoxygenated SS cells by a direct stimulation of the Ca2+ pump. PMA, moreover, inhibits deoxygenation-induced, charybdotoxin-sensitive K+ efflux in SS cells. This inhibition is partly indirect and explained by the reduced deoxygenation induced rise in cytosolic Ca2+ resulting from Ca2+ pump stimulation. However, a significant inhibition of the Ca2+-activated K+ channels (K(Ca) channels) by PMA can also be demonstrated when the channels are activated by Ca2+ plus ionophore, under conditions in which the Ca2+ pump is operating near its maximal extrusion rate, but swamped by Ca2+ plus ionophore. The data thus suggest a PKC alpha mediated phosphorylation both of the Ca2+ pump and of the K(Ca) channel or an auxiliary protein. PMID- 9357766 TI - Protein synthesis-dependent potentiation by thyroxine of antiviral activity of interferon-gamma. AB - We have studied the prenuclear signal transduction pathway by which thyroid hormone potentiates the antiviral activity of human interferon-gamma (IFN-gamma) in HeLa cells, which are deficient in thyroid hormone receptor (TR). The action of thyroid hormone was compared with that of milrinone, which has structural homologies with thyroid hormone. L-Thyroxine (T4), 3,5,3'-L-triiodothyronine (T3), and milrinone enhanced the antiviral activity of IFN-gamma up to 100-fold, a potentiation blocked by cycloheximide. The 5'-deiodinase inhibitor 6-n-propyl-2 thiouracil did not block the T4 effect. 3,3',5,5'-Tetraiodothyroacetic acid prevented the effect of T4 but not of milrinone. The effects of T4 and milrinone were blocked by inhibitors of protein kinases C (PKC) and A (PKA) and restored by PKC and PKA agonists; only the effect of T4 was blocked by genistein, a tyrosine kinase inhibitor. In separate models, milrinone was shown not to interact with nuclear TR-beta. T4 potentiation of the antiviral activity of IFN-gamma requires PKC, PKA, and tyrosine kinase activities but not traditional TR. PMID- 9357767 TI - Human umbilical vein and dermal microvascular endothelial cells show heterogeneity in response to PKC activation. AB - Changes in endothelial cell (EC) phenotype are central to the function of endothelium in inflammation. Although these events mainly occur in the microvasculature, previous studies have predominantly used large-vessel EC. Using enzyme-linked immunosorbent and flow cytometric assays, we compared the responses of human umbilical vein endothelial cells (HUVEC) and dermal microvascular endothelial cells (DMEC) to the activation of protein kinase C (PKC). Stimulation with phorbol 12,13-dibutyrate and more selective PKC agonists, including 12 deoxyphorbol 13-phenylacetate 20-acetate (dPPA), induced morphological changes and proliferation in both EC types. PKC activation induced a marked increase in Thy-1 expression on DMEC and only a moderate rise on HUVEC. Furthermore, heterogeneity in the induction of the adhesion molecules intercellular adhesion molecule 1, vascular cell adhesion molecule 1 IVCAM-1), and E-selectin between the two EC types following activation of PKC was demonstrated. In particular, E selectin and VCAM-1 were significantly upregulated on HUVEC but not DMEC. The data indicate that the PKC pathway is unlikely to be important for E-selectin and VCAM-1 expression in the microvasculature but are consistent with a role for PKC in angiogenesis. This diversity in signaling in response to PKC activation may depend on differential utilization of PKC isozymes and may facilitate specialized endothelial responses. PMID- 9357768 TI - Regulation of vascular angiotensin II receptors by EGF. AB - After vascular endothelial injury, angiotensin II (ANG II) plays a role in the resulting hypertrophic response, and expression of epidermal growth factor (EGF) is enhanced. Therefore, we tested the possibility that EGF regulates vascular ANG II action and receptor expression. Incubation of cultured aortic vascular smooth muscle cells (VSMC) with EGF (or basic fibroblast growth factor but not platelet derived growth factor isoforms) resulted in concentration-dependent (1-50 ng/ml EGF), time-dependent (>8 h), and reversible decreases in ANG II surface receptor density. For example, a 50% reduction was observed after exposure to 50 ng/ml EGF for 24 h. Incubation of cultured VSMC with 50 ng/ml EGF for 24 h resulted in a 77% reduction in ANG II-stimulated inositol phosphate formation. EGF not only prevented but also reversed ANG II receptor upregulation by 100 nM corticosterone. The specific tyrosine kinase inhibitor tyrphostin A48 (50 microM) reduced EGF-stimulated thymidine incorporation and EGF-stimulated phosphorylation of mitogen-activated protein kinase but did not prevent EGF from reducing ANG II receptor density. Neither pertussis toxin (100 ng/ml) nor downregulation of protein kinase C by phorbol myristate acetate (100 nM for 24 h) prevented EGF from reducing ANG II receptor density. In summary, EGF is a potent negative regulator of vascular ANG II surface receptor density and ANG II action by mechanisms that do not appear to include tyrosine phorphorylation, pertussis toxin-sensitive G proteins, or phorbol ester-sensitive protein kinase C. The possibility that EGF shifts the cell culture phenotype to one that exhibits reduced surface ANG II density cannot be eliminated by the present studies. PMID- 9357769 TI - Peroxide resistance of ER Ca2+ pump in endothelium: implications to coronary artery function. AB - We examined the effects of peroxide on the sarco(endo)plasmic reticulum Ca2+ (SERCA) pump in pig coronary artery endothelium and smooth muscle at three organizational levels: Ca2+ transport in permeabilized cells, cytosolic Ca2+ concentration in intact cells, and contractile function of artery rings. We monitored the ATP-dependent, azide-insensitive, oxalate-stimulated 45Ca2+ uptake by saponin-permeabilized cultured cells. Low concentrations of peroxide inhibited the uptake less effectively in endothelium than in smooth muscle whether we added the peroxide directly to the Ca2+ uptake solution or treated intact cells with peroxide and washed them before the permeabilization. An acylphosphate formation assay confirmed the greater resistance of the SERCA pump in endothelial cells than in smooth muscle cells. Pretreating smooth muscle cells with 300 microM peroxide inhibited (by 77 +/- 2%) the cyclopiazonic acid (CPA)-induced increase in cytosolic Ca2+ concentration in a Ca2+-free solution, but it did not affect the endothelial cells. Peroxide pretreatment inhibited the CPA-induced contraction in deendothelialized arteries with a 50% inhibitory concentration of 97 +/- 13 microM, but up to 500 microM peroxide did not affect the endothelium dependent, CPA-induced relaxation. Similarly, 500 microM peroxide inhibited the angiotensin-induced contractions in deendothelialized arteries by 93 +/- 2%, but it inhibited the bradykinin-induced, endothelium-dependent relaxation by only 40 +/- 13%. The greater resistance of the endothelium to reactive oxygen may be important during ischemia-reperfusion or in the postinfection immune response. PMID- 9357770 TI - Expression of smooth muscle myosin heavy chains and unloaded shortening in single smooth muscle cells. AB - The functional significance of the variable expression of the smooth muscle myosin heavy chain (SM-MHC) tail isoforms, SM1 and SM2, was examined at the mRNA level (which correlates with the protein level) in individual permeabilized rabbit arterial smooth muscle cells (SMCs). The length of untethered single permeabilized SMCs was monitored during unloaded shortening in response to increased Ca2+ (pCa 6.0), histamine (1 microM), and phenylephrine (1 microM). Subsequent to contraction, the relative expression of SM1 and SM2 mRNAs from the same individual SMCs was determined by reverse transcription-polymerase chain reaction amplification and densitometric analysis. Correlational analyses between the SM2-to-SM1 ratio and unloaded shortening in saponin- and alpha-toxin permeabilized SMCs (n = 28) reveal no significant relationship between the SM-MHC tail isoform ratio and unloaded shortening velocity. The best correlations between SM2/SM1 and the contraction characteristics of untethered vascular SMCs were with the minimum length attained following contraction (n = 20 and r = 0.72 for alpha-toxin, n = 8 and r = 0.78 for saponin). These results suggest that the primary effect of variable expression of the SM1 and SM2 SM-MHC tail isoforms is on the cell final length and not on shortening velocity. PMID- 9357771 TI - Partial cloning and characterization of Slc12a2: the gene encoding the secretory Na+-K+-2Cl- cotransporter. AB - The Slc12a2 gene encodes a widely expressed bumetanide-sensitive Na+-K+-2Cl- cotransporter that participates in various functions such as Cl- secretion and cell volume regulation. We isolated and characterized 75 kilobases of the murine gene encoding the cotransporter. The cotransport protein is encoded by 27 exons. Ribonuclease protection assay and primer extension demonstrated tissue-specific transcription initiation sites located within 270 base pairs upstream of the start codon. Nucleotide sequence analysis of the proximal 5'-flanking region revealed the presence of a weak TATA box, multiple Sp1/GC consensus sites, and the consensus sequence of a putative transcriptional initiator. Transfection of luciferase reporter gene constructs in mouse inner medullary collecting duct (mIMCD-3) cells confirmed the location of the minimal promoter within a 120-base pair fragment upstream of the cDNA. We also report the identification of an alternatively spliced variant of the cotransporter, expressed primarily in brain. This new spliced variant lacks exon 21, which encodes a 16-amino acid peptide located in the COOH-terminal tail of the protein. The absence of this exon causes the loss of the single protein kinase A consensus site of the cotransport protein. PMID- 9357772 TI - Block by MOPS reveals a conformation change in the CFTR pore produced by ATP hydrolysis. AB - ATP hydrolysis by the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel predicts that energy from hydrolysis might cause asymmetric transitions in the gating cycle. We found that 3-(N-morpholino)propanesulfonic acid (MOPS) blocked the open channel by binding to a site 50% of the way through the electrical field. Block by MOPS revealed two distinct states, O1 and O2, which showed a strong asymmetry during bursts of activity; the first opening in a burst was in the O1 state and the last was in the O2 state. Addition of a nonhydrolyzable nucleoside triphosphate prevented the transition to the O2 state and prolonged the O1 state. These data indicate that ATP hydrolysis by the nucleotide-binding domains drives a series of asymmetric transitions in the gating cycle. They also indicate that ATP hydrolysis changes the conformation of the pore, thereby altering MOPS binding. PMID- 9357773 TI - Cell cycle-dependent expression of a glioma-specific chloride current: proposed link to cytoskeletal changes. AB - We recently demonstrated expression of a novel, glioma-specific Cl- current in glial-derived tumor cells (gliomas), including stable cell lines such as STTG1, derived from a human anaplastic astrocytoma. We used STTG1 cells to study whether glioma Cl- channel (GCC) activity is regulated during cell cycle progression. Cells were arrested in defined stages of cell cycle (G0, G1, G1/S, S, and M phases) using serum starvation, mevastatin, hydroxyurea, demecolcine, and cytosine beta-D-arabinofuranoside. Cell cycle arrest was confirmed by measuring [3H]thymidine incorporation and by DNA flow cytometry. Using whole cell patch clamp recordings, we demonstrate differential changes in GCC activity after cell proliferation and cell cycle progression was selectively altered; specifically, channel expression was low in serum-starved, G0-arrested cells, increased significantly in early G1, decreased during S phase, and increased after arrest in M phase. Although the link between the cell cycle and GCC activity is not yet clear, we speculate that GCCs are linked to the cytoskeleton and that cytoskeletal rearrangements associated with cell division lead to the observed changes in channel activity. Consistent with this hypothesis, we demonstrate the activation of GCC by disruption of F-actin using cytochalasin D or osmotic cell swelling. PMID- 9357774 TI - Tunicamycin increases intracellular calcium levels in bovine aortic endothelial cells. AB - Tunicamycin is a nucleoside antibiotic that inhibits protein glycosylation and palmitoylation. The therapeutic use of tunicamycin is limited in animals because of its toxic effects, particularly in cerebral vasculature. Tunicamycin decreases palmitoylation of the endothelial isoform of nitric oxide synthase, stimulates nitric oxide synthesis, and increases the concentration of intracellular calcium ([Ca2+]i) in bovine aortic endothelial cells (B. J. Buckley and A. R. Whorton. FASEB J. 11: A110, 1997). In the present study, we investigated the mechanism by which tunicamycin alters [Ca2+]i using the Ca2+-sensitive dye fura 2. We found that tunicamycin increased [Ca2+]i without increasing levels of inositol phosphates. When cells were incubated in the absence of extracellular Ca2+, [Ca2+]i rapidly rose in response to tunicamycin, although a full response was not achieved. The pool of intracellular Ca2+ mobilized by tunicamycin overlapped with that mobilized by thapsigargin. Extracellular nickel blocked a full response to tunicamycin when cells were incubated in the presence of extracellular Ca2+. The effects of tunicamycin on [Ca2+]i were partially reversed by washing out the drug, and the remainder of the response was inhibited by removing extracellular Ca2+. These results indicate that tunicamycin mobilizes Ca2+ from intracellular stores in a manner independent of phospholipase C activation and increases the influx of Ca2+ across the plasma membrane. PMID- 9357775 TI - Ryanodine receptor expression is associated with intracellular Ca2+ release in rat parotid acinar cells. AB - The ryanodine receptor mediates intracellular Ca2+ mobilization in muscle and nerve, but its physiological role in nonexcitable cells is less well defined. Like adenosine 3',5'-cyclic monophosphate and inositol 1,4,5-trisphosphate, cyclic ADP-ribose (0.3-5 microM) and ADP (1-25 microM) produced a concentration dependent rise in cytosolic Ca2+ in permeabilized rat parotid acinar cells. Adenosine and AMP were less effective. Ryanodine markedly depressed the Ca2+ mobilizing action of the adenine nucleotides and forskolin in permeabilized cells and was likewise effective in depressing the action of forskolin in intact cells. Cyclic ADP-ribose-evoked Ca2+ release was enhanced by calmodulin and depressed by W-7, a calmodulin inhibitor. A fluorescently labeled ligand, 4,4-difluoro-1,3,5,7 tetramethyl-4-bora-3,4-diaza-s-indac ene-3-propionic acid-glycyl ryanodine, was synthesized to detect the expression and distribution of ryanodine receptors. In addition, ryanodine receptor expression was detected in rat parotid cells with a sequence highly homologous to a rat skeletal muscle type 1 and a novel brain type 1 ryanodine receptor. These findings demonstrate the presence of a ryanodine sensitive intracellular Ca2+ store in rat parotid cells that shares many of the characteristics of stores in muscle and nerve and may mediate Ca2+-induced Ca2+ release or a modified form of this process. PMID- 9357776 TI - The Ca2+-sensing receptor: a target for polyamines. AB - The Ca2+-sensing receptor (CaR) is activated at physiological levels of external Ca2+ (Ca(o)) but is expressed in a number of tissues that do not have well established roles in the control of Ca(o), including several regions of the brain and the intestine. Polyamines are endogenous polyvalent cations that can act as agonists for the CaR, as shown by our current studies of human embryonic kidney (HEK-293) cells transfected with the human CaR. Cellular parameters altered by polyamines included cytosolic free Ca2+ (Ca(i)), inositol phosphate production, and the activity of a nonselective cation channel. Spermine stimulated Ca(i) transients in CaR-transfected HEK cells, with a concentration producing a half maximal response (EC50) of approximately 500 microM in the presence of 0.5 mM Ca2+, whereas sustained increases in Ca(i) had an EC50 of approximately 200 microM. The order of potency was spermine > spermidine >> putrescine. Elevation of Ca(o) shifted the EC50 for spermine sharply to the left, with substantial stimulation below 100 microM. Addition of subthreshold concentrations of spermine increased the sensitivity of CaR-expressing HEK cells to Ca(o). Parathyroid hormone secretion from bovine parathyroid cells was inhibited by 50% in the presence of 200 microM spermine, a response similar to that elicited by 2.0 mM Ca(o). These data suggest that polyamines could be effective agonists for the CaR, and several tissues, including the brain, may use the CaR as a target for the actions of spermine and other endogenous polycationic agonists. PMID- 9357777 TI - ANG II AT1 and AT2 receptors both inhibit bFGF-induced proliferation of bovine adrenocortical cells. AB - Angiotensin II (ANG II) has long been known for its pressor and growth-promoting effects, which are both mediated by the AT1 receptor. By contrast, the AT2 receptor has recently been reported to mediate inhibition of proliferation through as yet undefined mechanisms. We report here that in bovine adrenal fasciculata cells ANG II by itself does not affect growth but inhibits basic fibroblast growth factor (bFGF)-induced DNA synthesis and blocks the cells in G1 phase. Consistent with this, ANG II inhibits cyclin D1 expression and cyclin D1 associated kinase activity. The antimitogenic effect of ANG II is partly mimicked by the AT2-selective agonist CGP-42112. It is also blocked partly and in an additive fashion by the AT1- and AT2-selective antagonists losartan and PD 123319, indicating the contribution of both receptor subtypes to this response. AT1-dependent antiproliferation is selectively blocked by the cyclooxygenase inhibitor indomethacin and restored by prostaglandin E2, whereas AT2-receptor mediated inhibition of growth is suppressed by the tyrosine phosphatase inhibitors orthovanadate and bpV(pic). Both pathways are, however, pertussis toxin sensitive. We hypothesize that, in fasciculata cells, the AT1 receptor inhibits bFGF-induced proliferation by stimulating prostaglandin synthesis, whereas the AT2 receptor mediates its effect through a pathway that requires protein tyrosine phosphatase activation. PMID- 9357778 TI - Male sex steroids are responsible for depressing macrophage immune function after trauma-hemorrhage. AB - Recent studies suggest beneficial effects of castration before soft tissue trauma and hemorrhagic shock on splenocyte immune functions. Nonetheless, it remains unknown whether this effect of testosterone depletion is limited to splenocytes or is a generalized effect on immune function. The present study was therefore carried out to determine whether androgen depletion before trauma-hemorrhage also has salutary effects on splenic and peritoneal macrophage as well as on Kupffer cell function, as indicated by interleukin (IL)-1 and IL-6 release. Male C3H/HeN mice were castrated or sham-castrated 2 wk before the experiment and were killed at 24 h after trauma-hemorrhage and resuscitation. Significant depression of macrophage IL-1 and IL-6 release was only observed in sham-castrated mice, as opposed to normal levels of cytokine release from castrated animals after trauma hemorrhage. In addition, only sham-castrated animals showed significantly increased levels of IL-6 release from Kupffer cells, which is believed to contribute to the systemic inflammatory response to trauma-hemorrhage. These observations suggest that the beneficial effects of androgen depletion before trauma-hemorrhage are not limited to splenocyte immune functions but are more global in nature. These results in surgically castrated animals suggest that androgen-blocking agents should be studied for their potential to reverse the immunodepression associated with trauma-hemorrhage. PMID- 9357779 TI - Role of water and electrolyte influxes in anoxic plasma membrane disruption. AB - The role of water and electrolyte influxes in anoxia-induced plasma membrane disruption was investigated using rabbit proximal tubule suspension. The results indicated that normal proximal tubule (PT) cells have a great capacity for expanding cell volume in response to water influx, whereas anoxia increases the susceptibility to water influx-induced disruption, and this was attenuated by glycine. However, resistance of anoxic plasma membranes to water influx-induced stress is not lost, although their mechanical strength was diminished, compared with normoxic membranes. Anoxic membranes did not disrupt under an intra-to extracellular osmotic difference as great as 150 mosM. Potentiating or attenuating water influx by incubating PT cells in hypotonic or hypertonic medium, respectively, during anoxia, did not affect anoxia-induced membrane disruption. After the transmembrane electrolyte concentration gradient was eliminated by a "intracellular" buffer or by permeabilizing the plasma membrane to molecules <4 kDa using alpha-toxin, anoxia still caused further membrane disruption that was prevented by glycine or low pH. These results demonstrate that 1) water or net electrolyte influxes are probably not a primary cause for anoxia-induced membrane disruption and 2) glycine could prevent the plasma membrane disruption during anoxia independently from its effect on transmembrane electrolyte or water influxes. The present data support a biochemical rather than a mechanical alteration of the plasma membrane as the underlying cause of membrane disruption during anoxia. PMID- 9357780 TI - Parathyroid hormone stimulates calcium influx and the cAMP messenger system in rat enterocytes. AB - Direct effects of parathyroid hormone (PTH) on calcium uptake by isolated rat duodenal cell preparations enriched in enterocytes were investigated. PTH significantly stimulated enterocyte 45Ca2+ influx in a time-dependent (1-10 min) manner and at all doses tested (2 x 10(-13) to 10(-7) M). The Ca2+ channel antagonists verapamil (10 microM) and nitrendipine (1 microM) completely blocked the stimulation of Ca2+ influx by the hormone (10(-8) M). PTH markedly increased cAMP levels in rat duodenal cells (88, 167, and 67%, after 1, 2, and 3 min, respectively). In agreement with these observations, forskolin (adenylate cyclase activator), dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP), and Sp-cAMPS (cAMP analogs) mimicked, whereas Rp-cAMPS (cAMP antagonist) suppressed PTH and DBcAMP activation of enterocyte calcium uptake. Furthermore, the effects of DBcAMP were abolished by nitrendipine. These results show direct rapid effects of PTH on duodenal cells' Ca2+ influx, which involve the activation of a dihydropyridine-sensitive Ca2+ influx pathway and the cAMP second messenger system. PMID- 9357781 TI - Adenosine stimulates Cl- channels of nonpigmented ciliary epithelial cells. AB - Ciliary epithelial cells possess multiple purinergic receptors, and occupancy of A1 and A2 adenosine receptors is associated with opposing effects on intraocular pressure. Aqueous adenosine produced increases in short-circuit current across rabbit ciliary epithelium, blocked by removing Cl- and enhanced by aqueous Ba2+. Adenosine's actions were further studied with nonpigmented ciliary epithelial (NPE) cells from continuous human HCE and ODM lines and freshly dissected bovine cells. With gramicidin present, adenosine (> or = 3 microM) triggered isosmotic shrinkage of the human NPE cells, which was inhibited by the Cl- channel blockers 5-nitro-2-(3-phenylpropylamino)benzoate (NPPB) and niflumic acid. At 10 microM, the nonmetabolizable analog 2-chloroadenosine and AMP also produced shrinkage, but not inosine, UTP, or ATP. 2-Chloroadenosine (> or = 1 microM) triggered increases of whole cell currents in HCE cells, which were partially reversible, Cl- dependent, and reversibly inhibited by NPPB. Adenosine (> or = 10 microM) also stimulated whole cell currents in bovine NPE cells. We conclude that occupancy of adenosine receptors stimulates Cl- secretion in mammalian NPE cells. PMID- 9357782 TI - Regulation of apolipoprotein A-I gene expression in Hep G2 cells depleted of Cu by cupruretic tetramine. AB - Studies were designed to examine the regulation of apolipoprotein (apo) A-I gene expression in Cu-depleted Hep G2 cells. The cupruretic chelator N,N'-bis(2 aminoethyl)-1,3-propanediamine 4 HCl (2,3,2-tetramine or TETA) was used to maintain a 77% reduction in cellular Cu in Hep G2 cells. After two passages of TETA treatment, the relative abundance of apoA-I mRNA was elevated 52%. In TETA treated cells, the rate of apoA-I mRNA decay measured by an actinomycin D chase study was accelerated 108%, and the synthesis of apoA-I mRNA determined by a nuclear runoff assay was enhanced 2.5-fold in TETA-treated cells. All of those changes could be reverted toward the control values with Cu supplementation for only 2 days. In transient transfection assays, a 26.7% increase in chloramphenicol O-acetyltransferase (CAT) activity for the reporter construct 256AI-CAT was observed in the treated cells. However, the ability of apoA-I regulatory protein 1 (ARP-1) to repress the CAT activity was not affected by the depressed Cu status. In addition, gel retardation experiments demonstrated that Cu depletion enhanced the binding of hepatocyte nuclear factor 4 (HNF-4) and other undefined nuclear factors to oligonucleotides containing site A, one of three regulatory sites of the apoA-I gene promoter. Moreover, the relative abundance of HNF-4 mRNA was increased 58% in the Cu-depleted cells. Thus the observed increase in apoA-I gene transcription may be mediated mostly by an elevated level of the regulatory factor, HNF-4. In summary, the present findings established the mechanism by which a depressed cellular Cu status can enhance apoA-I mRNA production and subsequently increase apoA-I synthesis. PMID- 9357783 TI - Beta-adrenergic regulation of constitutive nitric oxide synthase in cardiac myocytes. AB - Nitric oxide (NO) has been implicated in endogenous control of myocardial contractility. However, NO release has not yet been demonstrated in cardiac myocytes. Accordingly, endogenous NO production was measured with a porphyrinic microsensor positioned on the surface of individual neonatal or adult rat ventricular myocytes (n > 6 neonatal and adult cells per experiment). In beating neonatal myocytes, there was no detectable spontaneous NO release with each contraction. However, norepinephrine (NE; 0.25-1 microM) elicited transient NO release from beating neonatal (149 +/- 11 to 767 +/- 83 nM NO) and noncontracting adult (157 +/- 13 to 791 +/- 89 nM NO) cells. NO was released by adrenergic agonists with the following rank order of potency: isoproterenol (beta1beta2) > NE (alpha/beta1) > dobutamine (beta1) approximately epinephrine (alpha/beta1beta2) > tertbutylene (beta2); NO was not released by phenylephrine (alpha). NE-evoked NO release was reversibly blocked by N(G)-monomethyl-L arginine, trifluoperazine, guanosine 5'-O-(2-thiodiphosphate), and nifedipine but was enhanced by 3-isobutyl-1-methylxanthine (0.5 mM = 14.5 +/- 1.6%) and BAY K 8644 (10 microM = 11.9 +/- 1%). NO was also released by A-23187 (10 microM = 884 +/- 88 nM NO), guanosine 5'-O-(3-thiotriphosphate) (1 microM = 334 +/- 56 nM NO), and dibutyryl adenosine 3',5'-cyclic monophosphate (10-100 microM = 35 +/- 9 to 284 +/- 49 nM NO) but not by ATP, bradykinin, carbachol, 8-bromoguanosine 3',5' cyclic monophosphate, or shear stress. This first functional demonstration of a constitutive NO synthase in cardiac myocytes suggests its regulation by a beta adrenergic signaling pathway and may provide a novel mechanism for the coronary artery vasodilatation and enhanced diastolic relaxation observed with adrenergic stimulation. PMID- 9357784 TI - Physiological regulation of epithelial tight junctions is associated with myosin light-chain phosphorylation. AB - Tight junctions serve as the rate-limiting barrier to passive movement of hydrophilic solutes across intestinal epithelia. After activation of Na+-glucose cotransport, the permeability of intestinal tight junctions is increased. Because previous analyses of this physiological tight junction regulation have been restricted to intact mucosae, dissection of the mechanisms underlying this process has been limited. To characterize this process, we have developed a reductionist model consisting of Caco-2 intestinal epithelial cells transfected with the intestinal Na+-glucose cotransporter, SGLT1. Monolayers of SGLT1 transfectants demonstrate physiological Na+-glucose cotransport. Activation of SGLT1 results in a 22 +/- 5% fall in transepithelial resistance (TER) (P < 0.001). Similarly, inactivation of SGLT1 by addition of phloridzin increases TER by 24 +/- 2% (P < 0.001). The increased tight junction permeability is size selective, with increased flux of small nutrient-sized molecules, e.g., mannitol, but not of larger molecules, e.g., inulin. SGLT1-dependent increases in tight junction permeability are inhibited by myosin light-chain kinase inhibitors (20 microM ML-7 or 40 microM ML-9), suggesting that myosin regulatory light-chain (MLC) phosphorylation is involved in tight junction regulation. Analysis of MLC phosphorylation showed a 2.08-fold increase after activation of SGLT1 (P < 0.01), which was inhibited by ML-9 (P < 0.01). Thus monolayers incubated with glucose and myosin light-chain kinase inhibitors are comparable to monolayers incubated with phloridzin. ML-9 also inhibits SGLT1-mediated tight junction regulation in small intestinal mucosa (P < 0.01). These data demonstrate that epithelial cells are the mediators of physiological tight junction regulation subsequent to SGLT1 activation. The intimate relationship between tight junction regulation and MLC phosphorylation suggests that a critical step in regulation of epithelial tight junction permeability may be myosin ATPase-mediated contraction of the perijunctional actomyosin ring and subsequent physical tension on the tight junction. PMID- 9357786 TI - Kinetics of creatine kinase in an experimental model of low phosphocreatine and ATP in the normoxic heart. AB - To study the dependence of the forward flux of creatine kinase (CK) on its substrates and products we designed an acute normoxic model of steady-state depletion of phosphocreatine (PCr) and adenylate in the isovolumic acetate perfused rat heart. Various concentrations of PCr and ATP were induced by prior perfusion with 2 deoxy-D-glucose in the presence of insulin. The apparent rate constant (k(f)) and the forward CK flux were measured under metabolic and contractile steady state by progressive saturation-transfer 31P nuclear magnetic resonance (NMR). At high adenylate content CK flux was constant for a twofold reduction in PCr concentration ([PCr]); CK flux was 6.3 +/- 0.6 mM/s (vs. 6.5 +/- 0.2 mM/s in control) because of a doubling of k(f). Although, at the lowest ATP concentration and [PCr], CK flux was reduced by 50%, it nevertheless always remained higher than ATP synthesis estimated by parallel oxygen consumption measurement. NMR-measured flux was compared with the flux computed under the hypothesis of CK equilibrium. CK flux could not be fully predicted by the concentrations of CK metabolites. This is discussed in terms of metabolite and CK isozyme compartmentation. PMID- 9357785 TI - Evidence for heteromeric gap junction channels formed from rat connexin43 and human connexin37. AB - Homomeric gap junction channels are composed solely of one connexin type, whereas heterotypic forms contain two homomeric hemichannels but the six identical connexins of each are different from each other. A heteromeric gap junction channel is one that contains different connexins within either or both hemichannels. The existence of heteromeric forms has been suggested, and many cell types are known to coexpress connexins. To determine if coexpressed connexins would form heteromers, we cotransfected rat connexin43 (rCx43) and human connexin37 (hCx37) into a cell line normally devoid of any connexin expression and used dual whole cell patch clamp to compare the observed gap junction channel activity with that seen in cells transfected only with rCx43 or hCx37. We also cocultured cells transfected with hCx37 or rCx43, in which one population was tagged with a fluorescent marker to monitor heterotypic channel activity. The cotransfected cells possessed channel types unlike the homotypic forms of rCx43 or hCx37 or the heterotypic forms. In addition, the noninstantaneous transjunctional conductance-transjunctional voltage (Gj/Vj) relationship for cotransfected cell pairs showed a large range of variability that was unlike that of the homotypic or heterotypic form. The heterotypic cell pairs displayed asymmetric voltage dependence. The results from the heteromeric cell pairs are inconsistent with summed behavior of two independent homotypic populations or mixed populations of homotypic and heterotypic channels types. The Gj/Vj data imply that the connexin-to-connexin interactions are significantly altered in cotransfected cell pairs relative to the homotypic and heterotypic forms. Heteromeric channels are a population of channels whose characteristics could well impact differently from their homotypic counterparts with regard to multicellular coordinated responses. PMID- 9357788 TI - Predicted changes in concentrations of free and bound ATP and ADP during intracellular Ca2+ signaling. AB - High Ca2+ concentrations can develop near Ca2+ sources during intracellular signaling and might lead to localized regulation of Ca2+-dependent processes. By shifting the amount of Ca2+ and other cations associated with ATP, local high Ca2+ concentrations might also alter the substrate available for membrane associated and cytoplasmic enzymes. To study this, simultaneous equations were solved over a range of Ca2+ and Mg2+ concentrations to determine the general effects of Ca2+ on the concentrations of free and Ca2+- and Mg2+-bound forms of ATP. To obtain a more specific picture of the changes that might occur in smooth muscle cells, mathematical models of Ca2+ diffusion and regulation were used to predict the magnitude and time course of near-membrane Ca2+ transients and their effects on the free and bound forms of ATP near the membrane. The results of this work indicate that changes in free Ca2+ concentration over the range of 50 nM-100 microM would result in significant changes in free ATP concentration, MgATP concentration, and the CaATP-to-MgATP concentration ratio. PMID- 9357787 TI - Effect of osmolarity on LDL binding and internalization in hepatocytes. AB - The present study has been performed to elucidate a possible role of cell volume in low-density lipoprotein (LDL) binding and internalization (LDL(b+i)). As shown previously, increase of extracellular osmolarity (OSMe) and K+ depletion, both known to shrink cells, interfere with the formation of clathrin-coated pits and thus with LDL(b+i). On the other hand, alterations of cell volume have been shown to modify lysosomal pH, which is a determinant of LDL(b+i). LDL(b+i) have been estimated from heparin-releasable (binding) or heparin-insensitive (internalization) uptake of 125I-labeled LDL. OSMe was modified by alterations of extracellular concentrations of ions, glucose, urea, or raffinose. When OSMe was altered by varying NaCl concentrations, LDL(b+i) decreased (by 0.5 +/- 0.1%/mM) with increasing OSMe and LDL(b+i) increased (by 1.2 +/- 0.1%/mM) with decreasing OSMe, an effect mainly due to altered affinity; the estimated dissociation constant amounted to 20.6, 48.6, and 131.6 micro/ml at 219, 293, and 435 mosM, respectively. A 25% increase of OSMe increased cytosolic (by 0.46 +/- 0.03) and decreased lysosomal (by 0.14 +/- 0.02) pH. Conversely, a 25% decrease of OSMe decreased cytosolic (by 0.28 +/- 0.02) and increased lysosomal (by 0.17 +/- 0.02) pH. Partial replacement of extracellular Na+ with K+ had little effect on LDL(b+i), although it swelled hepatocytes and increased lysosomal and cytosolic pH. Hypertonic glucose, urea, or raffinose did not exert similar effects despite a shrinking effect of hypertonic raffinose. Monensin, which completely dissipates lysosomal acidity, virtually abolished LDL(b+i). In conclusion, the observations reveal a significant effect of ionic strength on LDL(b+i). The effect is, however, not likely to be mediated by alterations of cell volume or alterations of lysosomal pH. PMID- 9357789 TI - Gene expression of natriuretic peptide receptors in rats with DOCA-salt hypertension. AB - In our previous studies, we found that the atrial natriuretic peptide (ANP) binding and guanylyl cyclase activity of A-type natriuretic peptide receptors (NPR-A) were upregulated in renal papillae but downregulated in vascular tissues and glomeruli of rats with deoxycorticosterone acetate (DOCA)-salt hypertension [E. Nuglozeh, G. Gauquelin, R. Garcia, J. Tremblay, and E. L. Schiffrin. Am. J. Physiol. 259 (Renal Fluid Electrolyte Physiol. 28): F130-F137, 1990]. To further understand the molecular significance of these regulations, we measured the relative abundance of the transcripts of NPR-A and NPR-B by Northern blot in the aorta, mesenteric arteries, adrenal cortex, renal papillae, and lungs in DOCA salt hypertensive and control rats. In renal papillae we also examined the translation and transcription of NPR-A by ribosome loading and run-on assay. Compared with controls, the steady-state levels of mRNA for NPR-A were increased in the aorta and mesenteric arteries but were decreased in the adrenal cortex and renal papillae in DOCA-salt-treated rats. NPR-B mRNA was decreased in the aorta, mesenteric arteries, and adrenal cortex in hypertensive rats. In lungs the mRNA for both receptors was unchanged. Translation of NPR-A mRNA, as assessed by ribosome loading, was reduced in renal papillae. Transcriptional activity of its gene was not detectable in these tissues. Guanosine 3',5'-cyclic monophosphate levels generated by NPR-A in renal papillae and by NPR-A and NPR-B in the adrenal cortex, aorta, and mesenteric arteries of DOCA-salt-treated rats remained increased in hypertension. The higher NPR-A activity in the presence of a lower level of its mRNA in renal papillae and the higher NPR-B activity in the presence of a lower level of its mRNA in the vasculature, adrenal cortex, and lungs can alternatively be explained by receptor stabilization or increased receptor recycling. PMID- 9357790 TI - The role of Cl- channels in volume regulation in bovine pigmented epithelial cells. PMID- 9357791 TI - Binding sites for amiloride in intact epithelia. PMID- 9357793 TI - Intracellular mechanisms underlying prostaglandin F2alpha-stimulated phasic myometrial contractions. AB - These studies sought to test the hypothesis that prostaglandin F2alpha (PGF2alpha)-stimulated phasic myometrial contractions are characterized by the activation of the phosphatidylinositol-signaling pathway resulting in the generation of cytosolic calcium oscillations. For the experiments described in this report rat myometrial tissue was used, after the tissue was loaded with fura 2, to perform cytosolic calcium imaging studies and to perform computer digitalized in vitro isometric contraction studies. Consistent with the above hypothesis, the cytosolic calcium-imaging studies demonstrated PGF2alpha stimulated cytosolic calcium oscillations occurring simultaneously with phasic contractions. The in vitro isometric contraction studies confirmed that previously reported inhibitors of the phosphatidylinositol-signaling pathway and cytosolic calcium oscillation mechanisms resulted in significant inhibition of PGF2alpha-stimulated phasic myometrial contractions. In summary, these studies have provided substantial support for the hypothesis that PGF2alpha-stimulated phasic myometrial contractions are generated by intracellular signaling mechanisms involving activation of the phosphatidylinositol-signaling pathway and the production of cytosolic calcium oscillation-like phenomena. PMID- 9357792 TI - Insulin is degraded extracellularly in wounds by insulin-degrading enzyme (EC 3.4.24.56). AB - The exact mechanism by which insulin reverses impaired wound healing is unknown. Previous investigators have shown that insulin is degraded in experimental wounds, suggesting that the action of insulin may be locally modified. The following study corroborates these findings and identifies the major proteinase responsible for insulin degradation in wound fluid (WF). Adult male Fisher rats were wounded by subcutaneous implantation of polyvinyl alcohol sponges while under pentobarbital sodium anesthesia. WF and serum were collected on 1, 5, 10, and 14 days postinjury. Decreased insulin concentration in late WF correlated with an increased insulin-degrading activity. Multiple proteinases appear to participate in the overall degradation of insulin in WF. However, the primary enzyme responsible for insulin degradation in WF was characterized by immunoprecipitation and immunoblotting and identified as the neutral thiol dependent metalloproteinase, insulin-degrading enzyme (EC 3.4.24.56). Exogenous steroid administration caused a decrease in WF insulin-degrading activity. Glucagon and adrenocorticotrophin degradation was also observed, whereas minimal degradation of insulin-like growth factors I and II and epidermal growth factor was detected in WF. The ability to extracellularly degrade insulin may represent a unique mechanism for the regulation of this hormone's role in healing wounds. PMID- 9357794 TI - Role of epinephrine and norepinephrine in the metabolic response to stress hormone infusion in the conscious dog. AB - The role of epinephrine and norepinephrine in contributing to the alterations in hepatic glucose metabolism during a 70-h stress hormone infusion (SHI) was investigated in four groups of chronically catheterized (20-h-fasted) conscious dogs. SHI increased glucagon (approximately 5-fold), epinephrine (approximately 10-fold), norepinephrine (approximately 10-fold), and cortisol (approximately 6 fold) levels. Dogs received either all the hormones (SHI; n = 5), all the hormones except epinephrine (SHI-Epi; n = 6), or all the hormones except norepinephrine (SHI-NE; n = 6). In addition, six dogs received saline only (Sal). Glucose production (Ra) and gluconeogenesis were assessed after a 70-h hormone or saline infusion with the use of tracer ([3-(3)H]glucose and [U-(14)C]alanine) and arteriovenous difference techniques. SHI increased glucose levels (108 +/- 2 vs. 189 +/- 10 mg/dl) and Ra (2.6 +/- 0.2 vs. 4.1 +/- 0.3 mg x kg(-1) x min(-1)) compared with Sal. The absence of an increase in epinephrine markedly attenuated these changes (glucose and Ra were 140 +/- 6 mg/dl and 2.7 +/- 0.4 mg x kg(-1) x min(-1), respectively). Only 25% of the blunted rise in Ra could be accounted for by an attenuation of the rise in net hepatic gluconeogenic precursor uptake (0.9 +/- 0.1, 1.5 +/- 0.1, and 1.1 +/- 0.2 mg x kg(-1) x min(-1) for Sal, SHI, and SHI Epi, respectively). The absence of an increase in norepinephrine did not blunt the rise in arterial glucose levels, Ra, or net hepatic gluconeogenic precursor uptake (they rose to 195 +/- 21 mg/dl, 3.7 +/- 0.5 mg x kg(-1) x min(-1), and 1.7 +/- 0.2 mg x kg(-1) min(-1), respectively). In summary, during chronic SHI, the rise in epinephrine exerts potent stimulatory effects on glucose production principally by enhancing hepatic glycogenolysis, although the rise in circulating norepinephrine has minimal effects. PMID- 9357796 TI - Electrical stimulation induces fiber type-specific translocation of GLUT-4 to T tubules in skeletal muscle. AB - Insulin and contraction independently stimulate glucose transport in skeletal muscle. Whereas insulin activates glucose transport more in muscles composed of type I and IIa fibers, electrical stimulation increases glucose transport at least as much in type IIb fiber-enriched muscles despite the fact that the latter fiber type contains less GLUT-4 glucose transporters. The aim of the present study was to test the hypothesis that a greater GLUT-4 translocation to the cell surface may underlie the higher contraction-stimulated glucose transport in type IIb myofibers. Leg muscles from rats were stimulated in situ at 100 Hz (200 ms) each 2 s via the sciatic nerve over a period of 20 min while the contralateral leg was kept at rest. Muscle 2-[3H]deoxy-D-glucose uptake (2-DG) was measured in separated red gastrocnemius (RG, type I and IIa fibers) and white gastrocnemius (WG, type IIb fibers) muscles. Resting 2-DG uptake was greater in RG than WG. Electrical stimulation increased 2-DG uptake over resting values similarly in WG and RG. Fractions enriched with either plasma membranes, transverse (T) tubules, triads, or GLUT-4-enriched intracellular membranes were isolated from RG and WG using a recently developed subcellular fractionation procedure. Electrical stimulation similarly increased GLUT-4 protein content in plasma membranes of RG and WG, whereas it stimulated GLUT-4 translocation more (approximately 50%) in T tubules of WG than in RG. GLUT-4 content was not changed in triads of both muscle types. The increments in cell surface GLUT-4 protein levels were paralleled by significant reductions in the amount of the transporter in the intracellular membrane fractions of both muscle types (by 60% in RG and 56% in WG). It is concluded that electrically induced contraction stimulates GLUT-4 translocation more in T tubules of WG than RG. The physiological implications of this finding for glucose uptake by contracting RG and WG muscles is discussed. PMID- 9357795 TI - Regulation of glucose transporter GLUT-4 and hexokinase II gene transcription by insulin and epinephrine. AB - Transport of glucose across the plasma membrane by GLUT-4 and subsequent phosphorylation of glucose by hexokinase II (HKII) constitute the first two steps of glucose utilization in skeletal muscle. This study was undertaken to determine whether epinephrine and/or insulin regulates in vivo GLUT-4 and HKII gene transcription in rat skeletal muscle. In the first experiment, adrenodemedullated male rats were fasted 24 h and killed in the control condition or after being infused for 1.5 h with epinephrine (30 microg/ml at 1.68 ml/h). In the second experiment, male rats were fasted 24 h and killed after being infused for 2.5 h at 1.68 ml/h with saline or glucose (625 mg/ml) or insulin (39.9 microg/ml) plus glucose (625 mg/ml). Nuclei were isolated from pooled quadriceps, tibialis anterior, and gastrocnemius muscles. Transcriptional run-on analysis indicated that epinephrine infusion decreased GLUT-4 and increased HKII transcription compared with fasted controls. Both glucose and insulin plus glucose infusion induced increases in GLUT-4 and HKII transcription of twofold and three- to fourfold, respectively, compared with saline-infused rats. In conclusion, epinephrine and insulin may regulate GLUT-4 and HKII genes at the level of transcription in rat skeletal muscle. PMID- 9357797 TI - Potassium currents in ventricular myocytes from genetically diabetic rats. AB - Our previous study demonstrated the longer duration of action potential in ventricular myocytes from genetically diabetic WBN/Kob rats without change in calcium channel density compared with age-matched controls [Tsuchida, K., H. Watajima, and S. Otamo. Am. J. Physiol. 267 (Heart Circ. Physiol. 36): H2280 H2289, 1994]. In the present study we examined the alteration of potassium currents, especially transient outward current, in ventricular myocytes of genetically diabetic WBN/Kob rats. WBN/Kob rats gradually develop hyperglycemia with aging and show some similarity to non-insulin-dependent diabetes mellitus models, which differ from the insulin-dependent streptozotocin-treated rat model. The density of the intracellular calcium ion-independent transient outward current (I(to)) from 17- to 19-mo diabetic rat myocytes was significantly smaller than that from age-matched control rat myocytes. In addition, the density of I(to) from 17- to 19-mo rat myocytes was significantly less than that from 2-mo rat myocytes, suggesting that aging-induced alteration of I(to) was accelerated by the diabetic state. The steady-state inactivation curves of I(to), the recovery from I(to) inactivation, and the other outward currents were not significantly altered between diabetic myocytes and age-matched control myocytes. In conclusion, the prolonged duration of action potential from genetically diabetic rat myocytes is mainly due to the depressed I(to). PMID- 9357798 TI - Impaired adaptation of first-phase insulin secretion in postmenopausal women with glucose intolerance. AB - This study examined whether insulin secretion, insulin sensitivity, glucose effectiveness, and hepatic extraction of insulin are altered in subjects with impaired glucose tolerance (IGT). The frequently sampled intravenous glucose tolerance test was performed in postmenopausal women (age 63 yr, body mass index range 21.6-28.9 kg/m2) with IGT (n = 10) or normal glucose tolerance (NGT; n = 10). Insulin sensitivity (S(I)) was significantly lower in IGT than in NGT (P = 0.030). In contrast, insulin secretion was not significantly different between the two groups as determined by area under the curve for insulin and C-peptide, acute insulin response to glucose (AIR(G)), and glucose sensitivity of first phase (phi1) or of second-phase (phi2) insulin secretion. In NGT (r = -0.68, P = 0.029) but not in IGT (r = -0.05, not significant), S(I) correlated negatively with phi1. The B-cell "adaptation index" (S(I) x phi1) was lower in IGT than in NGT [83 +/- 25 vs. 171 +/- 29 min(-2)/(mmol/l), P = 0.042]. Also, the B-cell "disposition index" (S(I) times AIR(G)) was lower in IGT (83 +/- 25 10(-4) min( 1)) than in NGT (196 +/- 30 10(-4) min(-1), p = 0.011). In contrast, glucose effectiveness or hepatic extraction of insulin was not different between IGT and NGT. We conclude that postmenopausal women with IGT fail to adequately adapt to lowered S(I) by increasing first-phase insulin secretion. PMID- 9357800 TI - Caloric restriction increases HDL2 levels in rhesus monkeys (Macaca mulatta). AB - Caloric restriction (CR) prolongs the life of rodents and other small animals, but the benefits of CR for primates and people are as yet unknown, and mechanisms by which CR may slow aging remain unidentified. A study of rhesus monkeys, Macaca mulatta, is underway to determine if CR might prolong life span in primates and to evaluate potential mechanisms for life prolongation. Thirty rhesus monkeys in three age cohorts, restricted to 70% of ad libitum calorie intake for 6-7 yr, were compared with 30 controls. Plasma lipid, lipoprotein, and high-density lipoprotein (HDL) apolipoproteins and subfractions were measured and compared with weight, percent fat, glucose, and insulin level. CR caused decreased triglyceride levels in adult monkeys and increased levels of HDL2b, the HDL subfraction associated with protection from atherosclerosis. Multivariate statistical analyses showed that differences in lipid and lipoprotein levels occurring with CR could be accounted for, at least in part, by decreased body mass and improved glucose regulation. These studies have used a novel dietary modification paradigm in nonhuman primates focused on calorie reduction. Results suggest that CR, as mediated by its beneficial effect on body composition and glucose metabolism, could prolong human life by decreasing the incidence of atherosclerosis. PMID- 9357799 TI - Tissue triglycerides, insulin resistance, and insulin production: implications for hyperinsulinemia of obesity. AB - Obesity is associated with both insulin resistance and hyperinsulinemia. Initially hyperinsulinemia compensates for the insulin resistance and thereby maintains normal glucose homeostasis. Obesity is also associated with increased tissue triglyceride (TG) content. To determine whether both insulin resistance and hyperinsulinemia might be secondary to increased tissue TG, we studied correlations between TG content of skeletal muscle, liver, and pancreas and plasma insulin, plasma [insulin] x [glucose], and beta-cell function in four rat models with widely varying fat content: obese Zucker diabetic fatty rats, free feeding lean Wistar rats, hyperleptinemic Wistar rats with profound tissue lipopenia, and rats pair fed to hyperleptinemics. Correlation coefficients >0.9 (P < 0.05) were obtained among TG of skeletal muscle, liver, and pancreas and among plasma insulin, [insulin] x [glucose] product, and beta-cell function as gauged by basal, glucose-stimulated, and arginine-stimulated insulin secretion by the isolated perfused pancreas. Although these correlations cannot prove cause and effect, they are consistent with the hypothesis that the TG content of tissues sets the level of both insulin resistance and insulin production. PMID- 9357802 TI - Effects of exercise in diabetic rats before and during gestation on maternal and neonatal outcomes. AB - This study was designed to evaluate the effects of chronic endurance training on glucose and lipid homeostasis in diabetic mothers and their offspring. Female Sprague-Dawley rats were rendered diabetic (>20 mmol/l glucose) by streptozotocin and subdivided into three treatments (n = 10/group): exercise (20 m/min; 0% grade; 1 h/day; 5 days/wk) before and during gestation (EE), exercise before gestation with cessation on conception (ES), and sedentary before and during gestation (SS). Response of dams to a preconception and third trimester glucose tolerance test, litter number (EE = ES = SS = 3), and average litter size (EE = 9.7 +/- 1.5; ES = 9.0 +/- 1.5; SS = 8.3 +/- 0.3) did not differ among groups. Number of offspring remaining viable was significantly different among groups (EE = 17; ES = 0; SS = 14). Response to a glucose challenge and fasting glucose and insulin were different between the EE and SS pups. Exercise before and during gestation did not reduce the viability of offspring. Cessation of exercise during early pregnancy negatively affected offspring viability. PMID- 9357801 TI - Interleukin-8 can mediate acute-phase protein production by isolated human hepatocytes. AB - During the course of studies designed to identify the role of cytokines in the reprioritization of hepatic protein synthesis associated with cachexia we detected a hepatocyte-stimulating moiety in the supernatants of pancreatic cancer cells that was unrelated to interleukin (IL)-6. This study identifies that moiety as IL-8 and investigates the role of IL-8 in the induction of acute-phase protein production. The human pancreatic cancer cell line MIA PaCa-2 produced >1 ng/ml of IL-8 per 24 h, and supernatants from this cell line induced C-reactive protein (CRP) production from isolated human hepatocytes. Addition of neutralizing anti human IL-8 antibody to such supernatants produced almost complete inhibition of CRP production. The addition of recombinant human IL-8 to hepatocytes resulted in a dose-dependent increase in CRP, alpha1-acid glycoprotein, and alpha1 antichymotrypsin production and a decrease in the production of transferrin and prealbumin. This study demonstrates that recombinant or tumor-derived IL-8 can modulate acute-phase protein production from isolated human hepatocytes and from human hepatoma cells. PMID- 9357803 TI - Effects of estrogen and growth hormone on skeleton in the ovariectomized rat with hypophysectomy. AB - To investigate whether growth hormone (GH) and 17beta-estradiol (E2) replacement can prevent osteopenia induced by pituitary and ovarian hormone deficiency [by hypophysectomy and ovariectomy (HX+OV)], we administered relatively low doses of GH (2.3 IU x kg(-1) x day(-1)) and E2 (100 microg x kg(-1) x wk(-1)) in experiment 1 and relatively high doses of GH (13.5 IU x kg(-1) x day(-1)) and E2 (3,500 microg x kg(-1) x wk(-1)) in experiment 2 to 2-mo-old HX+OV Sprague-Dawley rats for 6 wk. Our data show that the HX+OV of rats results in diminished periosteal bone formation, longitudinal bone growth, and decreased cancellous bone volume. Administration of either the low or high dose of GH to these rats increased their systemic growth, serum levels of osteocalcin, and cortical bone formation. Either low or high doses of GH or E2 alone only partially prevent cancellous bone loss. However, the combined treatment of GH plus E2 resulted in an additive increase in the cancellous bone mass. We conclude that the additive effect of GH plus E2 on cancellous bone is attributed to the suppressive effect of E2 on bone resorption and the anabolic effect of GH on bone formation. PMID- 9357804 TI - Abnormal regulation of HGP by hyperglycemia in mice with a disrupted glucokinase allele. AB - Glucokinase (GK) catalyzes the phosphorylation of glucose in beta-cells and hepatocytes, and mutations in the GK gene have been implicated in a form of human diabetes. To investigate the relative role of partial deficiencies in the hepatic vs. pancreatic GK activity, we examined insulin secretion, glucose disposal, and hepatic glucose production (HGP) in response to hyperglycemia in transgenic mice 1) with one disrupted GK allele, which manifest decreased GK activity in both liver and beta-cells (GK+/-), and 2) with decreased GK activity selectively in beta-cells (RIP-GKRZ). Liver GK activity was decreased by 35-50% in the GK+/- but not in the RIP-GKRZ compared with wild type (WT) mice. Hyperglycemic clamp studies were performed in conscious mice with or without concomitant pancreatic clamp. In all studies [3-(3)H]glucose was infused to measure the rate of appearance of glucose and HGP during 80 min of euglycemia (Glc approximately 5 mM) followed by 90 min of hyperglycemia (Glc approximately 17 mM). During hyperglycemic clamp studies, steady-state plasma insulin concentration, rate of glucose infusion, and rate of glucose disappearance (Rd) were decreased in both GK+/- and RIP-GKRZ compared with WT mice. However, whereas the basal HGP (at euglycemia) averaged approximately 22 mg x kg(-1) x min(-1) in all groups, during hyperglycemia HGP was suppressed by only 48% in GK+/- compared with approximately 70 and 65% in the WT and RIP-GKRZ mice, respectively. During the pancreatic clamp studies, the ability of hyperglycemia per se to increase Rd was similar in all groups. However, hyperglycemia inhibited HGP by only 12% in GK+/-, vs. 42 and 45%, respectively, in the WT and RIP-GKRZ mice. We conclude that, although impaired glucose-induced insulin secretion is common to both models of decreased pancreatic GK activity, the marked impairment in the ability of hyperglycemia to inhibit HGP is due to the specific decrease in hepatic GK activity. PMID- 9357806 TI - Fatty acids reduce heparin-releasable LPL activity in cultured cardiomyocytes from rat heart. AB - Varying glucose and fatty acid (FA) concentrations in the medium of cultured cardiomyocytes from adult rat hearts were tested for effects on lipoprotein lipase (LPL) activity. Glucose (5.5, 11, and 25 mM in the culture medium for 18 22 h) had no effect on either heparin-releasable LPL (HR-LPL) or on cellular LPL (C-LPL) activities. When cardiomyocytes were cultured overnight with 60 microM oleate, HR-LPL activity was reduced to 20% of control, with no change in C-LPL activity or total C-LPL mass. Similar results (HR-LPL and C-LPL activities) were obtained with 60 microM concentrations of palmitate and myristate; linoleate and eicosapentaenoate did reduce C-LPL activity, but the decrease in HR-LPL activity was much greater. Oxfenicine, an FA oxidation inhibitor, did not alter the inhibitory effect of 60 microM oleate on HR-LPL. Short-term incubations (1 and 3 h) of cultured cardiomyocytes with 60 microM oleate did not displace LPL into the medium. Immunodetectable LPL on the cell surface of oleate-treated cultured cardiomyocytes was increased compared with control cells, but heparin treatment released the same amount of LPL mass that had reduced catalytic activity. PMID- 9357805 TI - Induction of NO and prostaglandin E2 in osteoblasts by wall-shear stress but not mechanical strain. AB - The nature of the stimulus sensed by bone cells during mechanical usage has not yet been determined. Because nitric oxide (NO) and prostaglandin (PG) production appear to be essential early responses to mechanical stimulation in vivo, we used their production to compare the responsiveness of bone cells to strain and fluid flow in vitro. Cells were incubated on polystyrene film and subjected to unidirectional linear strains in the range 500-5,000 microstrain (microepsilon). We found no increase in NO or PGE2 production after loading of rat calvarial or long bone cells, MC3T3-E1, UMR-106-01, or ROS 17/2.8 cells. In contrast, exposure of osteoblastic cells to increased fluid flow induced both PGE2 and NO production. Production was rapidly induced by wall-shear stresses of 148 dyn/cm2 and was observed in all the osteoblastic populations used but not in rat skin fibroblasts. Fluid flow appeared to act through an increase in wall-shear stress. These data suggest that mechanical loading of bone is sensed by osteoblastic cells through fluid flow-mediated wall-shear stress rather than by mechanical strain. PMID- 9357807 TI - Lipolytic suppression following carbohydrate ingestion limits fat oxidation during exercise. AB - This study determined if the suppression of lipolysis after preexercise carbohydrate ingestion reduces fat oxidation during exercise. Six healthy, active men cycled 60 min at 44 +/- 2% peak oxygen consumption, exactly 1 h after ingesting 0.8 g/kg of glucose (Glc) or fructose (Fru) or after an overnight fast (Fast). The mean plasma insulin concentration during the 50 min before exercise was different among Fast, Fru, and Glc (8 +/- 1, 17 +/- 1, and 38 +/- 5 microU/ml, respectively; P < 0.05). After 25 min of exercise, whole body lipolysis was 6.9 +/- 0.2, 4.3 +/- 0.3, and 3.2 +/- 0.5 micromol x kg(-1) x min( 1) and fat oxidation was 6.1 +/- 0.2, 4.2 +/- 0.5, and 3.1 +/- 0.3 micromol x kg( 1) x min(-1) during Fast, Fru, and Glc, respectively (all P < 0.05). During Fast, fat oxidation was less than lipolysis (P < 0.05), whereas fat oxidation approximately equaled lipolysis during Fru and Glc. In an additional trial, the same subjects ingested glucose (0.8 g/kg) 1 h before exercise and lipolysis was simultaneously increased by infusing Intralipid and heparin throughout the resting and exercise periods (Glc+Lipid). This elevation of lipolysis during Glc+Lipid increased fat oxidation 30% above Glc (4.0 +/- 0.4 vs. 3.1 +/- 0.3 micromol x kg(-1) x min(-1); P < 0.05), confirming that lipolysis limited fat oxidation. In summary, small elevations in plasma insulin before exercise suppressed lipolysis during exercise to the point at which it equaled and appeared to limit fat oxidation. PMID- 9357808 TI - Age-related decreases in stimulatory G protein-coupled adenylate cyclase activity in osteoblastic cells. AB - In this study we examined parathyroid hormone (PTH)-, forskolin (FSK)-, and cholera toxin (CTX)-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in rat osteoblastic cells (ROB) isolated from young (4 mo), mature (12 mo), and old (24-28 mo) male rats. Exposure to PTH increased cAMP accumulation in a concentration-dependent manner in all ROB cells examined. However, the maximum response in ROB from young rats was threefold greater than the maximum response in those from mature and old rats. Exposure to FSK also stimulated cAMP accumulation in a concentration-dependent manner, but there were no significant differences in responsiveness among ROB isolated from young, mature, and old rats. Exposure to CTX resulted in a dramatic concentration dependent increase in cAMP in ROB from young rats but only a modest increase in ROB from mature and old rats. PTH binding kinetics were similar in ROB from rats in each age group. These data suggest an age-related defect in stimulatory G protein coupling to adenylate cyclase, which contributes to decreased osteoblastic responsiveness to PTH. PMID- 9357809 TI - Myosin light-chain phosphorylation controls insulin secretion at a proximal step in the secretory cascade. AB - The aim of this study was to investigate how insulin secretion is controlled by phosphorylation of the myosin light chain (MLC). Ca2+-evoked insulin release from pancreatic islets permeabilized with streptolysin O was inhibited by different monoclonal antibodies against myosin light-chain kinase (MLCK) to an extent parallel to their inhibition of purified MLCK. Anti-MLCK antibody also inhibited insulin release caused by the stable GTP analog guanosine 5'-O-(3 thiodiphosphate), even at a substimulatory concentration (0.1 microM) of Ca2+. Free Ca2+ increased MLC peptide phosphorylation by beta-cell extracts in vitro. In contrast to the phosphorylation by purified MLCK or by calmodulin (CaM) kinase II, the activity partially remained with the beta-cell under nonstimulatory Ca2+ (0.1 microM) conditions. The MLCK inhibitor ML-9 inhibited the activity in the beta-cell with both substimulatory and stimulatory Ca2+, whereas KN-62, an inhibitor of CaM kinase II, only exerted an influence in the latter case. ML-9 decreased intracellular granule movement in MIN6 cells under basal and acetylcholine-stimulated conditions. We propose that MLC phosphorylation may modulate translocation of secretory granules, resulting in enhanced insulin secretion. PMID- 9357810 TI - Effects of aging on in vivo synthesis of skeletal muscle myosin heavy-chain and sarcoplasmic protein in humans. AB - A decline in muscle mass and contractile function are prominent features of the sarcopenia of old age. Because myosin heavy chain is an important contractile protein, it was hypothesized that synthesis of this protein decreases in sarcopenia. The fractional synthesis rate of myosin heavy chain was measured simultaneously with rates of mixed muscle and sarcoplasmic proteins from the increment of [13C]leucine in these proteins purified from serial needle biopsy samples taken from 24 subjects (age: from 20 to 92 yr) during a primed continuous infusion of L-[1-(13)C]leucine. A decline in synthesis rate of mixed muscle protein (P < 0.01) and whole body protein (P < 0.01) was observed from young to middle age with no further change with advancing age. An age-related decline of myosin heavy-chain synthesis rate was also observed (P < 0.01), with progressive decline occurring from young, through middle, to old age. However, sarcoplasmic protein synthesis did not decline with age. Myosin heavy-chain synthesis rate was correlated with measures of muscle strength (P < 0.05), circulating insulin-like growth factor I (P < 0.01), and dehydroepiandrosterone sulfate (P < 0.05) in men and women and free testosterone levels in men (P < 0.01). A decline in the synthesis rate of myosin heavy chain implies a decreased ability to remodel this important muscle contractile protein and likely contributes to the declining muscle mass and contractile function in the elderly. PMID- 9357811 TI - Differential effect of pp120 on insulin endocytosis by two variant insulin receptor isoforms. AB - The insulin receptor is expressed as two variably spliced isoforms that differ by the absence (isoform A) or presence (isoform B) of a 12-amino acid sequence encoded by exon 11 at the carboxy terminus of the alpha-subunit. Coexpression of the A isoform and pp120, a substrate of the insulin receptor tyrosine kinase, in NIH 3T3 fibroblasts increased receptor A-mediated insulin endocytosis and degradation by two- to threefold compared with cells expressing receptors alone. Because B is the predominant isoform in the liver and binds insulin with lower affinity than A, we have examined the effect of pp120 on receptor B-mediated endocytosis. In contrast to isoform A, the effect of pp120 on isoform B-mediated insulin internalization and degradation in stably transfected NIH 3T3 cells was minimal. PMID- 9357812 TI - Maximal aerobic capacity in African-American and Caucasian prepubertal children. AB - The purpose of this study was to examine differences in resting, submaximal, and maximal (VO2max) oxygen consumption (VO2) in African-American (n = 44) and Caucasian (n = 31) prepubertal children aged 5-10 yr. Resting VO2 was measured via indirect calorimetry in the fasted state. Submaximal VO2 and VO2max were determined during an all out, progressive treadmill exercise test appropriate for children. Dual-energy X-ray absorptiometry was used to determine total fat mass (FM), soft lean tissue mass (LTM), and leg soft LTM. Doubly labeled water was used to determine total energy expenditure (TEE) and activity energy expenditure (AEE). A significant effect of ethnicity (P < 0.01) was found for VO2max but not resting or submaximal VO2, with African-American children having absolute VO2max approximately 15% lower than Caucasian children (1.21 +/- 0.032 vs. 1.43 +/- 0.031 l/min, respectively). The lower VO2max persisted in African-American children after adjustment for soft LTM (1.23 +/- 0.025 vs. 1.39 +/- 0.031 l/min; P < 0.01), leg soft LTM (1.20 +/- 0.031 vs. 1.43 +/- 0.042 l/min; P < 0.01), and soft LTM and FM (1.23 +/- 0.025 vs. 1.39 +/- 0.031 l/min; P < 0.01). The lower VO2max persisted also after adjustment for TEE (1.20 +/- 0.02 vs. 1.38 +/- 0.0028 l/min P < 0.001) and AEE (1.20 +/- 0.024 vs. 1.38 +/- 0.028 l/min; P < 0.001). In conclusion, our data indicate that African-American and Caucasian children have similar rates of VO2 at rest and during submaximal exercise, but VO2max is approximately 15% lower in African-American children, independent of soft LTM, FM, leg LTM, TEE, and AEE. PMID- 9357813 TI - Intestinal growth is associated with elevated levels of glucagon-like peptide 2 in diabetic rats. AB - Glucagon-like peptide 2 (GLP-2) has recently been identified as a novel intestinal growth factor. Because experimental diabetes is associated with bowel growth, we examined the relationship between GLP-2 and intestinal growth in rats made diabetic by streptozotocin (STZ) injection and treated with or without insulin for 3 wk. Ileal concentrations of the intestinal proglucagon-derived peptides, i.e., glicentin + oxyntomodulin, and GLPs 1 and 2, were increased by 57 +/- 20% above those of controls in untreated STZ diabetes (P < 0.05-0.001). Similar increases in plasma concentrations of glicentin + oxyntomodulin (77 +/- 15% above controls, P < 0.01) and GLP-2 (91 +/- 32% above controls, P < 0.05) were seen in untreated STZ diabetes. Both wet and dry small intestinal weight increased by 74 +/- 20% above controls (P < 0.01) in STZ diabetes, and macromolecular analysis indicated parallel increases in both protein (P < 0.001) and lipid (P < 0.05) content. Villus height (P < 0.001) and crypt depth (P < 0.01) were also increased in untreated diabetic rat intestine. Insulin therapy prevented the changes in plasma GLP-2 and intestinal mass seen in untreated STZ diabetes. Thus STZ diabetes is associated with both increased production of GLP-2 and enhanced bowel weight, thereby suggesting a role for GLP-2 in diabetes associated bowel growth. PMID- 9357814 TI - A kinetic mass balance model for 1,5-anhydroglucitol: applications to monitoring of glycemic control. AB - The polyol 1,5-anhydroglucitol (AG) present in human plasma is derived largely from ingestion and is excreted unmetabolized. Reduction of plasma [AG] has been noted in diabetics and is due to accelerated excretion of AG during hyperglycemia. Plasma [AG] has therefore been proposed as a marker for glycemic control. A precise understanding of its utility relies on a quantitative understanding of the mass balance for AG. In this study, non-steady-state data from the literature were analyzed to develop a dynamic mass balance model for AG that is based on the two-compartment model proposed by Yamanouchi et al. [T. Yamanouchi, Y. Tachibana, H. Akanuma, S. Minoda, T. Shinohara, H. Moromizato, H. Miyashita, and I. Akaoka. Am. J. Physiol. 263 (Endocrinol. Metab. 26): E268-E273, 1992]. The data are consistent with a model in which exchange between tissue and plasma pools is rapid and in which the tissue compartment mass is two to three times the mass of the plasma compartment. According to model estimates, accelerated excretion of AG due to hyperglycemia can cause marked net depletion of total AG over a time scale of days. Recovery from a depleted state is slow because the total body capacity represents >5 wk of normal intake. Accordingly, AG monitoring should be able to indicate the presence of past glucosuric hyperglycemic episodes during a period of days to weeks, as well as provide information on the extent to which high deviations from the average plasma glucose concentration are operative. PMID- 9357815 TI - Epithelial cell growth and differentiation. IV. Controlled spatiotemporal expression of transgenes: new tools to study normal and pathological states. AB - The gut epithelium represents a dynamic, well-organized developmental system for examining self-renewal, differentiation, repair, and tumorigenesis. The apical pole of the enterocytes, the brush border, is composed of an array of well organized actin microfilaments that support the plasma membrane. Villin, one actin-binding protein that contributes to the assembly and dynamics of the microvillus bundle, exhibits special features such as restricted tissue specificity and early expression in the immature crypt cells. The regulatory elements of the villin gene are suitable to control the expression of transgenes in intestinal cells. Engineering genetically modified animals by classic transgenesis using the villin promoter or by gene targeting in the villin locus will allow the establishment of animal models that may recapitulate human intestinal disorders. PMID- 9357817 TI - Intestinal inflammation: a complex interplay of immune and nonimmune cell interactions. AB - Intestinal inflammation has traditionally been viewed as a process in which effector immune cells cause the destruction of other mucosal cells that behave as passive bystander targets. Progress in understanding the process of intestinal inflammation has led to a much broader and more integrated picture of the various mucosal components, a picture in which cytokines, growth factors, adhesion molecules, and the process of apoptosis act as functional mediators. Essentially all cellular and acellular components can exert immunelike activities, modifying the classical concept of selected immune cells acting on all other cells that has been the dogma of immunologically mediated tissue damage for decades. The existence of specialized communication pathways between epithelial cells and T cells is well documented, including abnormal epithelial cell-mediated T cell activation during inflammation. Mesenchymal cells contribute to fibrosis in the inflamed gut but are also responsible for retention and survival of leukocytes in the mucosa. In chronically inflamed intestine the local microvasculature displays leukocyte hyperadhesiveness, a phenomenon that probably contributes to persistence of inflammation. The extracellular matrix regulates the number, location, and activation of leukocytes, while metalloproteinases regulate the quantity and type of deposited matrix proteins. This evidence from the intestinal system, consolidated with the use of data from other organs and systems, reveals a rich network of reciprocal and finely orchestrated interactions among immune, epithelial, endothelial, mesenchymal, and nerve cells and the extracellular matrix. Although these interactions occur under normal conditions, the dysfunction of any component of this highly integrated mucosal system may lead to a disruption in communication and result in pathological inflammation. PMID- 9357816 TI - Cell adhesion and migration. I. Neutrophil adhesive interactions with intestinal epithelium. AB - In many inflammatory conditions of the gastrointestinal tract, disease activity and patient symptoms correlate with the histological finding of neutrophil (PMN) migration across the epithelium. PMN interactions with intestinal epithelium can influence epithelial functions ranging from barrier maintenance to electrolyte secretion. Additionally, PMN recruitment to the epithelium can be modulated by epithelial interactions with luminal enteric pathogens. Adhesive interactions between PMN and intestinal epithelial cells have been shown to be distinct from interactions of PMN with endothelia. In particular, PMN transepithelial migration is modulated by a distinct array of cytokines including interferon-gamma and interleukin-4 and requires the PMN beta2-integrin CD11b/CD18 but is independent of CD11a/CD18, selectins, and intercellular adhesion molecule 1. Additionally, an integral membrane protein termed CD47 has recently been shown to play an important role in PMN transepithelial migration at point(s) subsequent to initial adhesive interactions. This article provides a brief overview of PMN interactions with epithelia and their functional consequences in relation to inflammatory disease. PMID- 9357818 TI - In vitro analysis of rat intestinal wall movements at rest and during propagated contraction: a new method. AB - Intestinal wall motions are not easily studied and are frequently deduced from manometric and electromyographic measurements. This study aimed to establish a method of wall movement analysis based on an automatic technique of image processing. Segments of rat jejunum were fixed in an organ bath under isometric conditions. A real-time edge-detection algorithm was used to find the contours of the intestine using video imaging. After the measurement, a mapping of intestinal wall movements was performed based on diameter variations. In the 260 experiments without stimulation, intestinal wall activity was always detected. Propagated activity was found in 40% of the experiments and periodic wall motion in 60%, with 0.5-Hz activity found more frequently (41%) than 0.24-Hz activity (19%). These cyclic activities, related to intestinal slow waves, had their amplitude decreased by acetylcholine and were modified by vapreotide. Analysis of a propagated wave after cholinergic stimulation showed that it is characterized by an increase of the diameter of the intestine followed by a decrease. Moreover, this methodology allows analysis of the initiation of a propagated wave. PMID- 9357819 TI - Ca2+ channel blockade by verapamil inhibits GMCs and diarrhea during small intestinal inflammation. AB - The aim of this study was to investigate whether the blockade of L-type Ca2+ channels with verapamil suppresses giant migrating contractions (GMCs) and therefore diarrhea during small intestinal inflammation. Small intestinal inflammation was induced by infection with the nematode Trichinella spiralis. T. spiralis infection alone significantly increased the frequency of GMCs and decreased the frequency of phase III activity in the small intestine for 9 days. The increased frequency of GMCs was associated with diarrhea. Immunohistochemical staining with specific antibodies indicated that the number of neutrophils and mast cells increased significantly in the jejunal lamina propria during T. spiralis infection. Only the neutrophils increased significantly in the muscularis externa of the jejunum. Myeloperoxidase (MPO) activity increased significantly in the jejunal and ileal lamina propria. Daily verapamil administration during T. spiralis infection significantly reduced the frequency of GMCs and diarrhea but had no further significant effect on the already reduced frequency of phase III activity. Verapamil administration, however, did not reduce MPO activity or immunocyte infiltration in the jejunum or ileum. We conclude that blockade of L-type Ca2+ channels selectively reduces the frequency of GMCs and therefore diarrhea during small intestinal inflammation. The decreased frequency of GMCs is not secondary to a reduction in the inflammatory response. PMID- 9357820 TI - Hydrogen peroxide-induced liver cell necrosis is dependent on AP-1 activation. AB - To determine whether intracellular signaling events involved in apoptosis may also mediate necrosis, the role of the transcription factor AP-1 was investigated in a hepatoma cell model of cellular necrosis induced by oxidant stress. Treatment of the human hepatoma cell line HuH-7 with H2O2 caused dose-dependent necrosis as determined by light microscopy, fluorescent staining, and an absence of DNA fragmentation. H2O2 treatment led to increases in c-fos and c-jun mRNA levels, Jun nuclear kinase activity, and AP-1 DNA binding. AP-1 transcriptional activity measured with an AP-1-driven luciferase reporter gene was also increased. To determine whether this AP-1 activation contributed to H2O2-induced cell necrosis, HuH-7 cells were stably transfected with an antisense c-jun expression vector. Cells expressing antisense c-jun had decreased levels of AP-1 activation and significantly increased survival after H2O2 exposure. These data indicate that AP-1 activation occurs during oxidant-induced cell necrosis and contributes to cell death. Necrosis is therefore not always a passive process but may involve the activation of intracellular signaling pathways similar to those that mediate apoptosis. PMID- 9357821 TI - Fibronectin and cytokines increase JNK, ERK, AP-1 activity, and transin gene expression in rat hepatic stellate cells. AB - Cytokines, growth factors, and alterations in the extracellular matrix composition may play a role in maintaining hepatic stellate cells (HSC) in the activated state that is responsible for hepatic fibrogenesis. However, the signal transduction pathways that are stimulated by these factors in HSC remain to be fully elucidated. Recent evidence indicates that the mitogen-activated protein kinase (MAPK) family, including c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), plays an important role in the cellular response to stress. The aims of this study were to investigate whether fibronectin (FN) or the inflammatory cytokines interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) activate JNK, ERK, and AP-1 activity in HSC and induce the gene expression of the matrix metalloproteinase transin. Treatment of HSC with FN resulted in an up to 4.5-fold increase in ERK activity and a 2.1-fold increase in JNK activity. IL-1alpha and TNF-alpha produced up to a fourfold increase in JNK activity and a twofold increase in ERK activity. We then compared the effects of FN, IL-1alpha, and TNF-alpha on AP-1 activity and metalloproteinase mRNA induction. All three compounds increased AP-1 binding and promoter activity, and transin mRNA levels were increased 1.8-fold by FN, 2.2 fold by IL-1alpha, and 2.8-fold by TNF-alpha. Therefore, FN and inflammatory cytokines increase MAPK activity, stimulate AP-1 activity, and increase transin gene expression in HSC. Signal transduction pathways involving the MAPK family may play an important role in the regulation of matrix metalloproteinase expression by cytokines and FN in HSC. PMID- 9357822 TI - Oxidant-induced disruption of intestinal epithelial barrier function: role of protein tyrosine phosphorylation. AB - The effect of hydrogen peroxide (H2O2) on intestinal epithelial barrier function was examined in Caco-2 and T84 cell monolayers. H2O2 reduced transepithelial electrical resistance (TER) of Caco-2 and T84 cell monolayers. This decrease in TER was associated with a decrease in dilution potential and an increase in [3H]mannitol permeability, suggesting an H2O2-induced disruption of the paracellular junctional complexes. H2O2 administration also induced tyrosine phosphorylation of several proteins (at the molecular mass ranges of 50-90, 100 130, and 150-180 kDa) in Caco-2 cell monolayers. Phenylarsine oxide and sodium orthovanadate, inhibitors of protein tyrosine phosphatase, decreased TER and increased mannitol permeability and protein tyrosine phosphorylation (PTP). A low concentration of sodium orthovanadate also potentiated the effect of H2O2 on TER, dilution potential, mannitol permeability, and PTP. Pretreatment with genistein (30-300 microM) and tyrphostin (100 microM) inhibited the effect of H2O2 on TER, dilution potential, mannitol permeability, and PTP. These studies show that H2O2 increases the epithelial permeability by disrupting paracellular junctional complexes, most likely by a PTP-dependent mechanism. PMID- 9357823 TI - Epithelial cell kinase-B61: an autocrine loop modulating intestinal epithelial migration and barrier function. AB - Epithelial cell kinase (Eck) is a member of a large family of receptor tyrosine kinases whose functions remain largely unknown. Expression and regulation of Eck and its cognate ligand B61 were analyzed in the human colonic adenocarcinoma cell line Caco-2. Immunocytochemical staining demonstrated coexpression of Eck and B61 in the same cells, suggestive of an autocrine loop. Eck levels were maximal in preconfluent cells. In contrast, B61 levels were barely detectable in preconfluent cells and increased progressively after the cells reached confluence. Caco-2 cells cultured in the presence of added B61 showed a significant reduction in the levels of dipeptidyl peptidase and sucrase isomaltase mRNA, markers of Caco-2 cell differentiation. Cytokines interleukin 1beta (IL-1beta), basic fibroblast growth factor, IL-2, epidermal growth factor, and transforming growth factor-beta modulated steady-state levels of Eck and B61 mRNA and regulated Eck activation as assessed by tyrosine phosphorylation. Functionally, stimulation of Eck by B61 resulted in increased proliferation, enhanced barrier function, and enhanced restitution of injured epithelial monolayers. These results suggest that the Eck-B61 interaction, a target of regulatory peptides, plays a role in intestinal epithelial cell development, migration, and barrier function, contributing to homeostasis and preservation of continuity of the epithelial barrier. PMID- 9357824 TI - Hepatocyte nuclear factor-1alpha regulates transcription of the guanylin gene. AB - To study the molecular mechanisms controlling guanylin expression, we have cloned the mouse guanylin gene, including 2.7 kb of upstream sequence. We show that the first 133 base pairs (bp) of the upstream guanylin promoter are sufficient to drive near maximal (6-fold over basal) luciferase reporter gene expression in Caco-2 intestinal cells; at least 300 bp of upstream promoter are required for reporter gene expression in HT-29 intestinal cell lines. Using electromobility shift assays, we demonstrate that nuclear proteins bind to the hepatocyte nuclear factor-1 (HNF-1) consensus sequence in the guanylin promoter. The HNF-1 consensus sequence, located in the immediate 5' flanking region, is required for transcriptional activation of the guanylin gene in both intestinal cell lines. Mutagenesis of the HNF-1 consensus sequence abolishes transcriptional activation of guanylin promoter-luciferase reporter gene constructs. Cotransfection of these constructs with HNF-1alpha augments transcriptional initiation of the reporter gene. In contrast, HNF-1beta has no significant effect on transcription of the reporter gene. These experiments demonstrate that HNF-1alpha is an important regulatory element in the transcriptional activation of guanylin. PMID- 9357825 TI - cAMP increases liver Na+-taurocholate cotransport by translocating transporter to plasma membranes. AB - Adenosine 3',5'-cyclic monophosphate (cAMP), acting via protein kinase A, increases transport maximum of Na+-taurocholate cotransport within 15 min in hepatocytes (S. Grune, L. R. Engelking, and M. S. Anwer. J. Biol. Chem. 268: 17734-17741, 1993); the mechanism of this short-term stimulation was investigated. Cycloheximide inhibited neither basal nor cAMP-induced increases in taurocholate uptake in rat hepatocytes, indicating that cAMP does not stimulate transporter synthesis. Studies in plasma membrane vesicles showed that taurocholate uptake was not stimulated by the catalytic subunit of protein kinase A but was higher when hepatocytes were pretreated with cAMP. Immunoblot studies with anti-fusion protein antibodies to the cloned Na+-taurocholate cotransport polypeptide (Ntcp) showed that pretreatment of hepatocytes with cAMP increased Ntcp content in plasma membranes but not in homogenates. Ntcp was detected in microsomes, endosomes, and Golgi fractions, and cAMP pretreatment resulted in a decrease only in endosomal Ntcp content. It is proposed that cAMP increases transport maximum of Na+-taurocholate cotransport, at least in part, by translocating Ntcp from endosomes to plasma membranes. PMID- 9357826 TI - Alanine uptake activates hepatocellular chloride channels. AB - Cells involved in the retrieval and metabolic conversion of amino acids undergo significant increases in size in response to amino acid uptake. The resultant adaptive responses to cell swelling are thought to include increases in membrane K+ and Cl- permeability through activation of volume-sensitive ion channels. This viewpoint is largely based on experimental models of hypotonic swelling, but few mammalian cells experience hypotonic challenge in vivo. Here we have examined volume regulatory responses in a physiological model of cell-swelling alanine uptake in immortalized hepatocytes. Alanine-induced cell swelling was followed by a decrease in cell volume that was temporally associated with an increase in membrane Cl- currents. These currents were dependent both on alanine concentration and Na+, suggesting that currents were stimulated by Na+-coupled alanine uptake. Cl- currents were outwardly rectifying, exhibited an anion permeability sequence of I- > Br- > Cl-, and were inhibited by the Cl- channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid, features similar to those reported for a widely distributed class of volume-sensitive anion channels evoked by experimental hypotonic stress. These findings suggest that volume-sensitive anion channels participate in adaptive responses to amino acid uptake and provide such channels with a new physiological context. PMID- 9357827 TI - Characterization of the pharyngo-UES contractile reflex in humans. AB - Preliminary human studies suggest the presence of an upper esophageal sphincter (UES) contractile reflex triggered by pharyngeal water stimulation. The purposes of this study were to further characterize this reflex and determine the threshold volume for its activation. We studied 10 healthy young volunteers by manometric technique before and after topical pharyngeal anesthesia. UES pressure responses to various volumes and temperatures of water injected into the pharynx were elucidated. At a threshold volume, rapid-pulse and slow continuous pharyngeal water injection resulted in significant augmentation of UES pressure in all volunteers. Threshold volume for inducing UES contraction averaged 0.1 +/- 0.01 ml for rapid-pulse injection and was significantly smaller than that for slow continuous injection (1.0 +/- 0.2 ml). UES pressure increase duration averaged 16 +/- 4 s. Augmentation of UES resting tone by injection of water with three different temperatures was similar. This augmentation was abolished after topical anesthesia. Conclusions were that stimulation of the human pharynx by injection of minute amounts of water results in a significant increase in resting UES pressure: the pharyngo-UES contractile reflex. The magnitude of pressure increase due to activation of this reflex is not volume or temperature dependent. Loss of pharyngeal sensation abolishes this reflex. PMID- 9357828 TI - Effect of acute hyperglycemia on colorectal motor and sensory function in humans. AB - Increased use of laxatives and constipation are more common among people with diabetes mellitus than matched nondiabetic people in the same community. The mechanism of constipation in diabetes is unclear. Acute hyperglycemia was previously reported to reduce the gastrocolonic response. Our aim was to determine the effects of acute hyperglycemia on the colon compliance and motor response to feeding and on the sensory function of the colon and rectum in healthy human subjects. Eleven healthy individuals were studied under conditions of hyperglycemia (mean blood glucose 280 +/- 13 mg/dl) and euglycemia. We evaluated three parameters: 1) colonic motility and compliance by a multilumen manometry and barostatic balloon assembly in the descending colon (motility was studied during fasting and for 2 h postprandially); 2) perception of isobaric distensions of polyethylene balloons in the rectum and colon; and 3) rectal compliance. Initial tonic response to meal ingestion (0-5 min) was slightly lower during hyperglycemia (P = 0.3). However, colonic tone, motility, compliance, and sensation, as well as rectal compliance and sensation, were not significantly different under the conditions of euglycemia and acute hyperglycemia. In healthy individuals, acute hyperglycemia does not significantly change colonic or rectal motor functions or the perception of mechanosensory stimuli in the colon or rectum compared with euglycemia. These results do not support the hypothesis that hyperglycemia abolishes the colonic response to feeding. PMID- 9357829 TI - Morphometry and strain distribution in guinea pig duodenum with reference to the zero-stress state. AB - The aim of the present study is to determine the distribution of residual circumferential strains along the duodenum in anesthetized guinea pigs. A silicone elastomer was allowed to harden in the duodenal lumen under a pressure of 0.7 kPa. The duodenum was excised with the cast and photographed. The zero stress state was obtained by cutting rings of duodenum radially. The geometric configuration at the zero-stress state is of fundamental importance, because it is the basic state with respect to which the physical stresses and strains are defined. A basic piece of information is the way the tangent vector rotates from one end of the circumference to the other. In the duodenum at zero-stress state, the total rotation of the tangent from one tip to the other is -500 to -850 , with the lowest absolute value in the proximal duodenum. In other words, the duodenum usually turns itself inside out on changing from a loaded state to the zero-stress state. The serosal circumference, the duodenal wall thickness, and the ratio of wall thickness to mucosal circumference decreased in the distal direction. In the pressurized state, the serosal Cauchy strain was tensile and increased in the distal direction; the mucosal Cauchy strain was compressive in the proximal half of the duodenum and tensile in the distal half. The large circumferential residual strains must be taken into account in a study of physiological problems in which the stresses and strains are important, e.g., the bolus transport function. PMID- 9357830 TI - IGF-I induces collagen and IGFBP-5 mRNA in rat intestinal smooth muscle. AB - Insulin-like growth factor (IGF) binding protein 5 (IGFBP-5) mRNA was studied in intestines of rats with peptidoglycan-polysaccharide enterocolitis by Northern analysis and in situ hybridization. IGFBP-5 mRNA was increased 2.4 +/- 0.5-fold in inflamed rat colon compared with controls and was highly expressed in smooth muscle. Cultured rat intestinal smooth muscle cells were used to study the regulation of IGFBP-5 and type I collagen synthesis. IGF-I (100 ng/ml) increased IGFBP-5 mRNA (1.9 +/- 0.1-fold) and collagen type alpha1(I) mRNA (1.6 +/- 0.2 fold) in cultured smooth muscle cells. IGF-I induced a dose- and time-dependent increase in IGFBP-5 in conditioned medium by Western ligand blot and by immunoblot. IGF-I did not affect the IGFBP-5 mRNA decay rate after transcriptional blockade. Cycloheximide abolished IGFBP-5 mRNA. In conclusion, IGFBP-5 mRNA is expressed by intestinal smooth muscle and is increased during chronic inflammation. IGF-I increases IGFBP-5 and collagen mRNAs in intestinal smooth muscle cells. PMID- 9357831 TI - Responsiveness to growth factors in aortic vascular smooth muscle cells from rats with cirrhosis. AB - Hemodynamic changes in cirrhosis may be associated with alterations in aortic vascular smooth muscle cell (AVSMC) function. The present study compared the proliferative response to serum and growth factors in cirrhotic and control AVSMC. Serum from cirrhotic rats, cirrhotic cell lysates, and the conditioned medium of cultured cirrhotic AVSMC induced an increase in [3H]thymidine incorporation in control but not in cirrhotic AVSMC. Platelet-derived growth factor-beta (PDGF-BB) induced a greater increase in [3H]thymidine incorporation in cirrhotic than in control cells. [3H]thymidine incorporation induced by cirrhotic conditioned medium was blocked by anti-PDGF antibody. Immunoblot studies showed that the anti-PDGF antibody recognized a 30-kDa protein in the conditioned medium of cirrhotic AVSMC culture, a protein corresponding to PDGF. Binding studies of PDGF-BB indicated a twofold increase in receptor density in cirrhotic AVSMC with no alteration in affinity for PDGF-BB. We conclude that an increased responsiveness of cirrhotic AVSMC to the PDGF could contribute to alterations in AVSMC and muscle cell tone that may play a role in the hemodynamic changes in cirrhosis. PMID- 9357833 TI - Regulation of hepatic enzymes and insulin levels in offspring of rat dams fed a reduced-protein diet. AB - We have hypothesized that permanent changes caused by poor growth during early development due to maternal malnutrition may be exacerbated by overnutrition of offspring in later life. To test this hypothesis, rats were exposed to a maternal 20% protein diet or an isocaloric 8% protein diet during fetal and postnatal life. All offspring were weaned onto laboratory chow. At 6 wk, rats were fed laboratory chow or a highly palatable diet (high fat and high calorie with adequate protein) and studied at 12 wk after a 48-h fast. The highly palatable diet resulted in excess weight gain and higher plasma insulin levels in all animals. Plasma insulin concentrations were significantly increased in male offspring of dams fed a reduced-protein diet compared with male offspring of dams fed an adequate-protein diet, but no differences were observed between the female offspring. The key hepatic enzymes of glucose homeostasis programmed in offspring of protein-restricted rat dams retained the ability to respond to overnutrition during adult life. In these offspring, however, the enzymes were regulated around a "set point" that was different from that in the controls. PMID- 9357832 TI - Molecular mechanisms for the growth factor action of gastrin. AB - We have previously observed that gastrin has a cholecystokinin B (CCK-B) receptor mediated growth-promoting effect on the AR42J rat pancreatic acinar cell line and that this effect is paralleled by induction of expression of the early response gene c-fos. We undertook these experiments to elucidate the mechanism for induction of c-fos and the linkage of this action to the trophic effects of gastrin. Gastrin (0.1-10 nM) dose dependently induced luciferase activity in AR42J cells transfected with a construct consisting of a luciferase reporter gene coupled to the serum response element (SRE) of the c-fos promoter. This effect was blocked by the specific CCK-B receptor antagonist D2 but not by the specific CCK-A receptor antagonist L-364,718 or by pertussis toxin, indicating that gastrin targets the SRE via specific CCK-B receptors through a mechanism independent of Gi. Inhibition of protein kinase C (PKC) either by prolonged (24 h) exposure of the cells to the phorbol ester 12-O-tetradecanoylphorbol 13 acetate (100 nM) or by incubation with the selective inhibitor GF-109203X (3.5 microM) resulted in an 80% reduction in luciferase activity. Similar results were observed in the presence of the specific extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor PD-98059 (50 microM). We measured ERK2 activity in AR42J cells via in-gel kinase assays and observed that gastrin (1 pM-100 nM) induced ERK2 enzyme activity in a dose-dependent manner. Addition of GF-109203X and PD-98059, either alone or in combination, produced, respectively, partial and total inhibition of gastrin-induced ERK2 activity. Gastrin induction of ERK2 activity also resulted in a threefold increase in the transcriptional activity of Elk-1, a factor known to bind to the c-fos SRE and to be phosphorylated and activated by ERK2. PD-98059 blocked the growth-promoting effect of gastrin on the AR42J cells, demonstrating that this effect depends on activation of MEK. Our data lead us to conclude that the trophic actions of gastrin are mediated by ERK2 induced c-fos gene expression via PKC-dependent and -independent pathways. PMID- 9357834 TI - Control of rate and extent of the proliferative response after partial hepatectomy. AB - To examine the role of the early changes occurring in the liver within the first hours after a partial hepatectomy and in an attempt to demonstrate the involvement of subsequent regulatory mechanisms, the size of the remnant liver was modified at various times and by different surgical techniques. Male Wistar rats were submitted to a two-thirds "temporary partial hepatectomy" produced by a 3-h occlusion of the pedicle of the anterior lobes protected by local hypothermia. Various indexes of cell proliferation ([3H]thymidine uptake and 5 bromo-2'-deoxyuridine and proliferating cell nuclear antigen labeling) were not increased despite a c-myc expression as high as that observed after a two-thirds partial hepatectomy. The temporary partial hepatectomy and a sham operation induced modifications of the hepatocytes, allowing rapid DNA synthesis after a subsequent two-thirds partial hepatectomy. After this initial nonspecific response, the extent of the regenerative response is determined according to the size of the liver mass present approximately from the 10th to the 18th hour after the initial stimulus. For instance, when a one-third partial hepatectomy was converted into a two-thirds partial hepatectomy at the 10th hour, the DNA synthesis at the 24th hour reached the value observed after a straightforward two thirds partial hepatectomy. Inversely, the regenerative response was significantly reduced when additional liver lobes were connected to neck vessels between the 14th and the 18th hour after a two-thirds partial hepatectomy. In conclusion, the actual liver mass present during the period corresponding to mid- to late G1 appears to control the magnitude of the proliferative response, which is not the simple consequence of the early changes following a partial hepatectomy. PMID- 9357835 TI - Mechanism of inhibition of Na+-glucose cotransport in the chronically inflamed rabbit ileum. AB - In a rabbit model of chronic ileal inflammation, we previously demonstrated that coupled NaCl absorption was reduced because of an inhibition of Cl-/HCO3- but not Na+/H+ exchange on the brush-border membrane (BBM) of villus cells. In this study we determined the alterations in Na+-stimulated glucose [Na+-O-methyl-D-glucose (Na+-OMG)] absorption during chronic ileitis. Na+-OMG uptake was reduced in villus cells from the chronically inflamed ileum. Na+-K+-adenosinetriphosphatase (ATPase), which provides the favorable Na+ gradient for this cotransporter in intact cells, was found to be reduced also. However, in villus cell BBM vesicles from the inflamed ileum Na+-OMG uptake was reduced as well, suggesting an effect at the level of the cotransporter itself. Kinetic studies demonstrated that Na+ OMG uptake in the inflamed ileum was inhibited by a decrease in the maximal rate of uptake for OMG without a change in the affinity. Analysis of steady-state mRNA and immunoreactive protein levels of this cotransporter demonstrates reduced expression. Thus Na+-glucose cotransport was inhibited in the chronically inflamed ileum, and the inhibition was secondary to a decrease in the number of cotransporters and not solely secondary to an inhibition of Na+-K--ATPase or altered affinity for glucose. PMID- 9357836 TI - Glucagon-like peptide-1 inhibits gastric emptying via vagal afferent-mediated central mechanisms. AB - Exogenous administration of glucagon-like peptide-1-(7-36) amide (GLP-1), an insulinotropic hormone, inhibits gastric emptying and acid secretion in humans. The role of GLP-1 as a regulator of gastric function is elusive. In gastric fistula rats, vagal afferent denervation and peripheral administration of the GLP 1 receptor antagonist exendin-(9-39) amide enhanced emptying of a glucose meal, whereas intracerebroventricular exendin was ineffective. The rate of saline emptying was attenuated by peripheral as well as by central administration of GLP 1, and pretreatment with exendin by the respective routes reversed the inhibition by GLP-1. Vagal afferent denervation abolished the central and peripheral action of GLP-1 on gastric emptying. Neither peripheral cholinergic nor adrenergic blockade altered the delay of methyl cellulose meal emptying by intracisternal GLP-1 injection. Acid secretion in conscious pylorus-ligated rats was inhibited by intracisternal GLP-1 administration, whereas systemic GLP-1 was ineffective. These results support the notion that GLP-1 receptors participate in the central and peripheral regulation of gastric function. Furthermore, vagal afferent nerves mediate the inhibitory action of GLP-1 on gastric motor function. GLP-1 may be a candidate brain-gut peptide that acts as a physiological modulator of gastric function. PMID- 9357838 TI - Caerulein pancreatitis increases mRNA but reduces protein levels of rat pancreatic heat shock proteins. AB - We have recently reported that preconditioning through hyperthermia induces expression of pancreatic heat shock proteins (HSPs) and protects against caerulein pancreatitis. Here, we investigate caerulein-mediated effects on pancreatic HSPs without prior hyperthermia. Caerulein time and dose dependently increased pancreatic mRNA levels of the constitutive isoform of HSP70 (HSC70). However, pancreatic HSC70 protein levels were decreased, as were HSP60 protein levels. Also, in contrast to hyperthermia preconditioning, caerulein did not induce measurable levels of mRNA or protein of the inducible isoform of HSP70. Thus the pancreas reacts to different kinds of stress (hyperthermia vs. hyperstimulation) with differential induction of HSP mRNAs. Clearly, hyperthermia leads to induction of HSP protein expression, whereas caerulein treatment does not. Therefore, our current study further supports the idea that hyperthermia induced protection against caerulein pancreatitis may be mediated through increased protein levels of pancreatic HSPs. It is further tempting to hypothesize that failure to appropriately increase HSP protein levels in response to high doses of caerulein might be a factor in the development of pancreatitis. PMID- 9357837 TI - Acute experimental colitis decreases colonic circular smooth muscle contractility in rats. AB - Distal colitis decreases the contractility of the underlying circular smooth muscle. We examined how time after injury and lesion severity contribute to the decreased contractility and how colitis alters the calcium-handling properties of the affected muscle. Distal colitis was induced in rats by intrarectal administration of 4% acetic acid. Contractile responses to acetylcholine, increased extracellular potassium, and the G protein activator NaF were determined for circular muscle strips from sham control and colitic rats at days 1, 2, 3, 7, and 14 postenemas. Acetylcholine stimulation of tissues from day 3 colitic rats was performed in a zero calcium buffer, in the presence of nifedipine, and after depletion of intracellular stores of calcium. The colitis was graded macroscopically as mild, moderate, or severe. Regardless of agonist, maximal decrease in force developed 2 to 3 days posttreatment, followed by a gradual return to control by day 14. The inhibitory effect of colitis on contractility increased with increasing severity of inflammation. Limiting extracellular calcium influx had a greater inhibitory effect on tissues from colitic rats; intracellular calcium depletion had a greater inhibitory effect on tissues from control animals. The data suggest that both lesion severity and time after injury affect the contractile response of circular smooth muscle from the inflamed distal colon. Impaired utilization of intracellular calcium may contribute to the decreased contractility. PMID- 9357839 TI - Effects of a panel of dietary lectins on cholecystokinin release in rats. AB - We previously showed that soybean lectin (SBL) releases cholecystokinin (CCK) and have now asked whether other dietary lectins have this effect and if extracellular calcium is involved. Lectins and vehicle were first infused into the duodenum of anesthetized rats. The CCK response to vehicle was 3.1 +/- 0.6 pmol/l (P < 0.05 vs. basal). SBL and peanut lectin (PNL) (84 microg/ml) significantly increased plasma CCK concentrations from 2.0 +/- 0.4 pmol/l to a maximum of 8.4 +/- 0.5 pmol/l (P < 0.01 vs. vehicle, mean +/- SE) and from 1.9 +/ 0.5 to 7.0 +/- 0.6 pmol/l (P < 0.05 vs. vehicle, mean +/- SE), respectively. Wheat germ lectin (WGL) (840 microg) also increased plasma CCK levels from 1.5 +/ 0.3 pmol/l to a maximum of 9.7 +/- 1.3 pmol/l (P < 0.05 vs. vehicle, mean +/- SE). Corresponding increases in pancreatic protein output occurred. Broad bean lectin (BBL) had no effect on either parameter. Dose-dependent responses were seen with SBL, PNL, and WGL (1, 10, and 100 microg/ml) in perifused rat intestinal cells. These responses were abolished in calcium-free medium and in the presence of the competing sugars of the lectins. Therefore, SBL, PNL, and WGL, which bind to motifs including N-acetyl-D-galactosamine, galactose, and N acetylglucosamine, respectively, released CCK, but BBL, which binds to mannose and glucose, did not. Ingestion of lectins may have major CCK-mediated effects on gastrointestinal function and growth. PMID- 9357841 TI - Dynamics of esophageal bolus transport in healthy subjects studied using multiple intraluminal impedancometry. AB - The dynamics of a bolus transport through the esophagus are largely unexplored. To study this physiological process, we applied multiple intraluminal impedancometry in 10 healthy subjects. Three different protocols were used: 1) liquid bolus administered with subject supine, 2) liquid bolus with subject upright, or 3) semisolid bolus with subject supine. Transit of different parts of a bolus (bolus head, body, and tail) was analyzed at different anatomic segments, namely the pharynx and the proximal, middle, and distal thirds of the esophagus. A characteristic pattern of bolus transport was seen in all subjects. Impedance changes related to air were observed preceding the bolus head. The bolus head propelled significantly faster than did the bolus body and tail. Pharyngeal bolus transit was significantly faster than esophageal bolus transit. Within the esophagus, bolus propulsion velocity gradually decreased. Bolus transport was significantly accelerated in the upright position and delayed with increase of bolus viscosity. In conclusion, the dynamics of a bolus transport from the pharynx into the stomach are complex. It varies within both different anatomic segments and different parts of the bolus and depends on bolus characteristics and test conditions. The spatial and temporal resolution of a bolus transport can be obtained by the impedance technique. PMID- 9357840 TI - Use of measurements of ethanol absorption from stomach and intestine to assess human ethanol metabolism. AB - Controversy exists concerning the site (stomach vs. liver) and magnitude of first pass metabolism of ethanol. We quantitated gastric and total ethanol absorption rates in five male subjects and utilized these measurements to evaluate first pass metabolism. Gastric emptying of ethanol (0.15 g/kg) was determined via a gamma camera and gastric absorption from the ratio of gastric ethanol to [14C]polyethylene glycol. Gastric absorption accounted for 30% and 10% of ethanol administered with food and water, respectively. With food, estimated gastric mucosal ethanol concentrations fell from 19 to 5 mM over 2 h. Calculations using these concentrations and kinetic data for gastric alcohol dehydrogenase showed <2% of the dose underwent gastric metabolism. Application of observed ethanol absorption rates to a model of human hepatic ethanol metabolism indicated that only 30% and 4% of the dose underwent first-pass metabolism when administered with food and water, respectively. We conclude that virtually all first-pass ethanol metabolism occurs in the liver and first-pass metabolism accounts for only a small fraction of total clearance. PMID- 9357842 TI - Demonstration of changes in fetal liver erythropoiesis using echo-planar magnetic resonance imaging. AB - This study investigated the variation in magnetic resonance characteristics of the fetal liver during a time of changing erythropoietic function. Echo-planar imaging was carried out in 25 normal pregnant women at 20 and 26 wk gestation. The signal intensity from regions of the fetal liver, background image, and maternal back muscle and the highest signal intensity from the maternal spinal cord were measured and compared with the signal intensity of amniotic fluid. Data are expressed as ratios, in arbitrary units (median pixel values; interquartile range shown in parentheses), and analyzed with the use of Wilcoxon's signed-rank test. At 20 wk, the signal intensity ratio of liver to amniotic fluid was 0.309 (0.231-0.365). At 26 wk, the ratio was 0.544 (0.429-0.616). The change was highly significant (P < 0.0001). No change in the signal intensity ratios of amniotic fluid compared with other measured parameters was noted. These data are consistent with known changes in fetal liver erythropoiesis occurring between 20 and 26 wk gestation and have potential use in early noninvasive physiological assessment of the fetus. PMID- 9357843 TI - Glucose and glutamine provide similar proportions of energy to mucosal cells of rat small intestine. AB - The objectives of this study were to establish a reliable method for quantifying glycolytic flux in intestinal epithelial cells, to determine the proportion of energy provided to small intestine epithelial cells by glucose vs. glutamine, and to determine whether there was an energetic advantage to having both substrates present simultaneously. There was substantial retention of 3H in alanine and lactate when [2-(3)H]glucose was used as tracer for quantifying glycolysis, and the magnitude of the 3H retention was influenced by the presence of other substrates and metabolites. Detritiation was at least 99% complete, however, when [3-(3)H]glucose was used as tracer in this system and the tritium was recovered as 3H2O. Glycolytic flux was six- to sevenfold higher in cells of the proximal than distal small intestine but was not significantly different for young adult (4 mo) vs. aged adult (24 mo) rats. Net ATP production from exogenous substrates was higher when both glucose and glutamine were present simultaneously than when either substrate was present alone, and glucose was calculated to provide 50-60% of the net ATP produced from these two substrates. Most of the energy produced from glucose was produced via the anaerobic metabolic pathways (78% for glucose alone, 95% with glucose and glutamine). Net energy production was calculated to be 10% lower in cells from aged animals than in those from young animals, since CO2 production from these major substrates was lower in cells from aged animals. PMID- 9357844 TI - Surfactant metabolic consequences of overexpression of GM-CSF in the epithelium of GM-CSF-deficient mice. AB - Granulocyte macrophage colony-stimulating factor (GM-CSF) is a regulator of surfactant metabolism because GM-CSF-deficient mice have abnormally slow clearance and catabolism of saturated phosphatidylcholine (Sat PC) and surfactant protein (SP)-A in airspaces and lung tissue. Overexpression of GM-CSF only in respiratory epithelial cells of mice deficient in GM-CSF using the SP-C promotor (GM-/-,SP-C-GM+/+) resulted in increased type II cell numbers and normalization of alveolar Sat PC pool sizes. Metabolic measurements demonstrated that incorporation of radiolabeled choline and palmitate was increased more than twofold, but the amount of radiolabeled Sat PC that accumulated in airspaces relative to the amount incorporated was decreased by 50% relative to normal GM+/+ mice. The clearance of dipalmitoylphosphatidylcholine and SP-B from the airspaces was more rapid for GM-/-,SP-C-GM+/+ mice than for GM+/+ mice. Loss of Sat PC and SP-B from the lungs (alveolar plus lung tissue) was similar in the two strains of mice. The normal surfactant pools in the GM-/-,SP-C-GM+/+ mice were achieved by the net effects of increases in type II cell numbers, increased incorporation, decreased accumulation, and increased reuptake rates for surfactant components, demonstrating the multiple effects of GM-CSF on surfactant metabolism. PMID- 9357845 TI - GM-CSF enhances lung growth and causes alveolar type II epithelial cell hyperplasia in transgenic mice. AB - The human surfactant protein (SP)-C gene promoter was used to direct expression of mouse granulocyte macrophage colony-stimulating factor (GM-CSF; SP-C-GM mice) in lung epithelial cells in GM-CSF-replete (GM+/+) or GM-CSF null mutant (GM-/-) mice. Lung weight and volume were significantly increased in SP-C-GM mice compared with GM+/+ or GM-/- control mice. Immunohistochemical staining demonstrated marked type II cell hyperplasia, and immunofluorescent labeling for proliferating cell nuclear antigen was increased in type II cells of SP-C-GM mice. Abundance of type II cells per mouse lung was increased three- to fourfold in SP-C-GM mice compared with GM+/+ and GM-/- mice. GM-CSF increased bromodeoxyuridine labeling of isolated type II cells in vitro. Type II cells, alveolar macrophages, and endothelial and bronchiolar epithelial cells were stained by antibodies to the GM-CSF receptor alpha-subunit in both GM+/+ mice and GM-CSF gene-targeted mice that are also homozygous for the SP-C-GM transgene. High levels of GM-CSF expression in type II cells of transgenic mice increased lung size and caused type II cell hyperplasia, demonstrating an unexpected role for the molecule in the regulation of type II cell proliferation and differentiation. PMID- 9357846 TI - Hyperoxia inhibits fetal rat lung fibroblast proliferation and expression of procollagens. AB - The direct effects of hyperoxia on collagen production by fetal lung fibroblasts are unknown and would be important to the understanding of the molecular mechanisms involved in bronchopulmonary dysplasia in premature infants. We studied the effect of hyperoxia on 1) proliferation, 2) mRNA levels for type I and III procollagens, and 3) net collagen production in primary cultures of fetal rat lung fibroblasts. Fibroblasts from 19-day-old rat fetuses (term is 22 days) were obtained. Test plates were incubated in hyperoxia and controls in room air for varying time periods. Cell viability in both conditions was >97% as determined by trypan blue exclusion. Fibroblast proliferation in nonconfluent cultures was found to be significantly reduced with exposure to hyperoxia (P < 0.001). Steady-state levels of type I and III procollagen mRNAs, analyzed on Northern blots hybridized to [32P]cDNA probes, were significantly decreased in hyperoxia (P < 0.01). This effect was noted as early as 4 h of exposure to hyperoxia and persisted for 5 days. There was a significant inverse correlation between duration of exposure to O2 and steady-state levels of mRNA for alpha1(I) procollagen (r = -0.904) and alpha1(III)-procollagen (r = -0.971). There were no significant changes in steady-state levels of beta-actin mRNA. We also found a significant decrease in collagen synthesis in hyperoxia-exposed fibroblasts (P < 0.05). We conclude that hyperoxia directly effects a reduction in fetal lung fibroblast proliferation and net collagen production at a pretranslational level. PMID- 9357847 TI - The nature of leukocyte shape changes in the pulmonary capillaries. AB - The size discrepancy between leukocytes [white blood cells (WBCs)] and pulmonary capillaries requires WBCs to deform. We investigated the persistence of this deformation on cells leaving the capillary bed and the role played by the cytoskeleton. Isolated rabbit lungs were perfused in situ via the pulmonary artery with effluent fractions collected from the left ventricle. Washout curves from cell counts in each fraction confirmed that WBCs are preferentially retained over erythrocytes. WBC deformation present on exit from the circulation was compared with that present after recovery in paired fractions, fixed either immediately or 60 min later. These cells were compared with cells recovered from the capillary in perfused fixative or fixed in peripheral blood. Our results show that leukocyte deformation persisted after the cells exited the pulmonary circulation. This deformation was associated with minimal submembranous F-actin staining, and microtubule distribution and cell polarization were unchanged. We conclude that cytoskeletal changes that occur during WBC deformation in the pulmonary capillaries are minimal and differ from those known to occur in actively migrating cells during chemotaxis. PMID- 9357848 TI - Exosurf enhances adenovirus-mediated gene transfer to alveolar type II cells. AB - We assessed the role of surfactant replacement mixtures in the enhancement of adenovirus-mediated gene transfer to pulmonary epithelial cells both in vitro and in vivo. A549 cells, a pulmonary epithelium-derived adenocarcinoma cell line, were incubated with either media alone or media containing 10 microg phospholipid/ml Exosurf or Infasurf for 50 min followed by addition of a replication-deficient adenovirus (E1-deleted) expressing the luciferase reporter gene [AdCMV-Luc; 10 plaque-forming units (PFU)/cell] for 4 h. Pretreatment with Exosurf, but not Infasurf, at 37 degrees C, but not at 4 degrees C, enhanced luciferase activity in A549 cells 24 h later by 156% (P < 0.01). Intratracheal instillation of AdCMV-Luc (2 x 10(9) PFU) into rats resulted in luciferase expression mainly in alveolar macrophages and to a smaller extent in alveolar type II (ATII) cells 24 h later. However, when the AdCMV-Luc instillation was preceded by Exosurf (250 microl; 25 mg/ml), a 10-fold increase in ATII cell luciferase activity was noted. Preincubation of cultured ATII cells with Exosurf also enhanced their transfection by AdCMV-Luc by 515% (P < 0.001). The results of these studies provide a new strategy for targeting ATII cells for gene delivery. PMID- 9357849 TI - Rhinovirus infection of primary cultures of human tracheal epithelium: role of ICAM-1 and IL-1beta. AB - Exacerbations of asthma are often associated with respiratory infection caused by rhinoviruses. To study the effects of rhinovirus infection on respiratory epithelium, a primary target for respiratory viruses, human rhinovirus (HRV)-2 and HRV-14 were infected to primary cultures of human tracheal epithelial cells. Viral infection was confirmed by showing that viral titers of supernatants and lysates from infected cells increased with time and by polymerase chain reaction. HRV-2 and HRV-14 infections upregulated the expression of intercellular adhesion molecule-1 (ICAM-1) mRNA, the major rhinovirus receptor, on epithelial cells, and they increased the production of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha in supernatants. Antibodies to ICAM-1 inhibited HRV 14 infection of epithelial cells and decreased the production of cytokines after HRV-14 infection, but they did not alter HRV-2 infection-induced production ofcytokines. IL-1beta upregulated ICAM-1 mRNA expression and increased susceptibility to HRV-14 infection, whereas other cytokines failed to alter ICAM 1 mRNA expression. Furthermore, a neutralizing antibody to IL-1beta significantly decreased viral titers of supernatants and ICAM-1 mRNA expression after HRV-14 infection, but a neutralizing antibody to TNF-alpha was without effect. Immunohistochemical studies revealed that both HRV-14 infection and IL-1beta increased ICAM-1 expression on cultured epithelial cells. These findings imply that HRV-14 infection upregulated ICAM-1 expression on epithelial cells through increased production of IL-1beta, thereby increasing susceptibility to infection. These events may be important for amplification of airway inflammation after viral infection in asthma. PMID- 9357850 TI - Involvement of the ICE family of proteases in silica-induced apoptosis in human alveolar macrophages. AB - Exposure to silica dust can result in lung inflammation that may progress to fibrosis for which there is no effective clinical treatment. The mechanisms involved in the development of pulmonary silicosis have not been well defined; however, most current evidence implicates a central role for alveolar macrophages in this process. We have previously demonstrated that fibrotic agents, such as asbestos and silica, induce apoptosis in human alveolar macrophages. The goal of this study was to identify molecular events in the silica-induced apoptotic process to better understand the mechanism by which fibrotic agents may be inducing apoptosis in human alveolar macrophages. To elucidate the possible mechanism by which silica causes apoptosis, we investigated the involvement of the interleukin-converting enzyme (ICE) family of proteases. Human alveolar macrophages were treated with silica in vitro and were examined for the involvement of ICE, Ich-1L, and cpp32beta in silica-induced apoptosis. Pretreatment of cells with 10 microM of the ICE inhibitor z-Val-Ala-Asp fluoromethyl ketone and the cpp32beta inhibitor Asp-Glu-Val-Asp-fluoromethyl ketone completely blocked silica-induced apoptosis. Additionally, an increased formation of the active p20 fragments of ICE and Ich-1L as well as degradation of the inactive zymogen form of cpp32beta protein were observed in silica-treated human alveolar macrophages, indicating activation of these proteases. Furthermore, degradation of the nuclear protein poly(ADP-ribose) polymerase was observed within 2 h of silica treatment. These results suggest that silica induced apoptosis involves activation of the ICE family of proteases and is the first step in elucidating the intracellular mechanism of particulate-induced apoptosis in human alveolar macrophages. PMID- 9357851 TI - Oxidant stress regulates basal endothelin-1 production by cultured rat pulmonary endothelial cells. AB - Endothelin-1 (ET-1) is a pluripotent mediator that modulates vascular tone and influences the inflammatory response. Patients with inflammatory lung disorders frequently have elevated circulating ET-1 levels. Because these pathophysiological conditions generate reactive oxygen species that can regulate gene expression, we investigated whether the level of oxidant stress influences ET-1 production in cultured rat pulmonary arterial endothelial cells (RPAEC). Treatment with the antioxidant 1,3-dimethyl-2-thiourea (10 mM) or the iron chelator deferoxamine (1.8 microM) doubles basal ET-1 release. Conversely, exposing cells to H2O2 generated by glucose and glucose oxidase (0.1-10 mU/ml) for 4 h causes a concentration-dependent decrease in ET-1 release. This effect occurs at concentrations of glucose oxidase that do not affect [3H]leucine incorporation or specific 51Cr release from RPAEC. Catalase prevents the decrease in ET-1 synthesis caused by glucose and glucose oxidase. Glucose and glucose oxidase decrease not only ET-1 generation but also ET-1 mRNA as assessed by semiquantitative polymerase chain reaction. Our results indicate that changes in oxidative stress can either up- or downregulate basal ET-1 generation by cultured pulmonary endothelial cells. PMID- 9357852 TI - Role of protein kinase C in calcium sensitization during muscarinic stimulation in airway smooth muscle. AB - Muscarinic receptor stimulation increases Ca2+ sensitivity, i.e., the amount of force produced at a constant submaximal cytosolic Ca2+ concentration ([Ca2+]i), in permeabilized smooth muscle preparations. It is controversial whether this increase in Ca2+ sensitivity is in part mediated by protein kinase C (PKC). With the use of a beta-escin permeabilized canine tracheal smooth muscle (CTSM) preparation, the effect of four putative PKC inhibitors [calphostin C, chelerythrine chloride, a pseudosubstrate inhibitor for PKC [PKC peptide-(19 31)], and staurosporine] on Ca2+ sensitization induced by acetylcholine (ACh) plus GTP was determined. Preincubation with each of the inhibitors did not affect subsequent Ca2+ sensitization induced by muscarinic receptor stimulation in the presence of a constant submaximal [Ca2+]i, neither did any of these compounds reverse the increase in Ca2+ sensitivity induced by ACh plus GTP. Administration of a 1,2-diacylglycerol analog, 1-oleoyl-2-acetyl-sn-glycerol, did not induce Ca2+ sensitization at a constant submaximal [Ca2+]i. Thus we found no evidence that PKC mediates increases in Ca2+ sensitivity produced by muscarinic receptor stimulation in permeabilized CTSM. PMID- 9357853 TI - Ascorbate deficiency and oxidative stress in the alveolar type II cell. AB - The objective of this study was to determine the impact of limited ascorbate (Asc) availability on type II cell sensitivity to oxidant stress. Guinea pigs were fed diets with or without Asc for 18 days, and type II cells were isolated. Although lung Asc was decreased by 90% in deficient animals (scorbutic), type II cell Asc was decreased by 50%. Upon treatment with 250 microM H2O2, the necrotic injury was twofold greater in scorbutic cells compared with control cells. With 100 microM H2O2 treatment, apoptotic injury was twofold greater in scorbutic cells compared with control cells. Although there was less necrotic injury in cells exposed to 95% O2, the scorbutic cells were more sensitive than control cells. Asc pretreatment protected against necrosis and apoptosis. The Asc analog isoascorbate provided partial protection and suggested that part of the protection was not chemical detoxification but was Asc specific. We conclude that limited Asc availability resulted in a functional type II cell but a cell more sensitive to oxidant-induced injury. PMID- 9357854 TI - Differential induction of c-fos, c-jun, and apoptosis in lung epithelial cells exposed to ROS or RNS. AB - Reactive oxygen (ROS) or nitrogen (RNS) species can affect epithelial cells to cause acute damage and an array of pulmonary diseases. The goal of this study was to determine patterns of early response gene expression and functional end points of exposure to nitric oxide (NO.), H2O2, or peroxynitrite (ONOO-) in a line of rat lung epithelial (RLE) cells. Our focus was on c-fos and c-jun protooncogenes, as these genes play an important role in proliferation or apoptosis, possible end points of exposure to reactive metabolites in lung. Our data demonstrate that NO. generated by spermine 1,3-propanediamine N-14-[1-(3-aminopropyl)-2-hydroxy-2 nitrosohydrazino]-butyl] or S-nitroso-N-acetylpenicillamine as well as H2O2 cause increased c-fos and c-jun mRNA levels, nuclear proteins, and complexes binding the activator protein-1 recognition sequence in RLE cells. These agents also lead to apoptosis and increased membrane permeability. In contrast, exogenously administered ONOO- or 3-morpholinosydnonimine do not induce protooncogenes or apoptosis in RLE cells despite nitration oftyrosines. We conclude that ROS and RNS can elicit distinct molecular and phenotypic responses in a target cell of pulmonary disease. PMID- 9357855 TI - Impairment of cation transport in A549 cells and rat alveolar epithelial cells by hypoxia. AB - A reduced cation reabsorption across the alveolar epithelium decreases water reabsorption from the alveoli and could diminish clearing accumulated fluid. To test whether hypoxia restricts cation transport in alveolar epithelial cells, cation uptake was measured in rat lung alveolar type II pneumocytes (AII cells) in primary culture and in A549 cells exposed to normoxia and hypoxia. In AII and A549 cells, hypoxia caused a PO2-dependent inhibition of the Na-K pump, of Na-K 2Cl cotransport, and of total and amiloride-sensitive 22Na uptake. Nifedipine failed to prevent hypoxia-induced transport inhibition in both cell types. In A549 cells, the inhibition of the Na-K pump and Na-K-2Cl cotransport occurred within approximately 30 min of hypoxia, was stable >20 h, and was reversed by 2 h of reoxygenation. There was also a reduction in cell membrane-associated Na-K ATPase and a decrease in Na-K-2Cl cotransport flux after full activation with calyculin A, indicating a decreased transport capacity. [14C]serine incorporation into cell proteins was reduced in hypoxic A549 cells, but inhibition of protein synthesis with cycloheximide did not reduce ion transport. In AII and A549 cells, ATP levels decreased slightly, and ADP and the ATP-to-ADP ratio were unchanged after 4 h of hypoxia. In A549 cells, lactate, intracellular Na, and intracellular K were unchanged. These results indicate that hypoxia inhibits apical Na entry pathways and the basolateral Na-K pump in A549 cells and rat AII pneumocytes in culture, indicating a hypoxia-induced reduction of transepithelial Na transport and water reabsorption by alveolar epithelium. If similar changes occur in vivo, the impaired cation transport across alveolar epithelial cells might contribute to the formation of hypoxic pulmonary edema. PMID- 9357857 TI - Rhinovirus stimulation of interleukin-8 in vivo and in vitro: role of NF-kappaB. AB - Neutrophil infiltration is a well-documented early event in the pathogenesis of rhinovirus (RV) infections. To further understand the mechanisms responsible for this neutrophilia, we determined whether interleukin (IL)-8 was present at sites of experimental RV infection in vivo and characterized the mechanism(s) by which RV stimulates IL-8 production in vitro. IL-8 was readily detectable in the nasal washings of all normal volunteers and did not increase with sham nasal inoculation. In contrast, RV infection caused a significant additional increase in nasal IL-8, the levels of which peaked 48-72 h after virus inoculation. RV was a potent stimulator of IL-8 protein production by A549 epithelial-like cells, MRC 5 fibroblasts, and normal human bronchial epithelial cells in vitro. This induction was associated with a significant increase in IL-8 mRNA accumulation and gene transcription. RV also stimulated IL-8 promoter-driven luciferase activity. This stimulation was significantly decreased by mutation of the nuclear factor (NF)-IL-6 site and was completely abrogated by mutation of the NF-kappaB site in this promoter. In addition, NF-kappaB-DNA binding activity was rapidly induced in RV-infected cells. This inducible binding was made up of p65 and, to a lesser extent, p50 NF-kappaB moieties. These studies demonstrate that IL-8 is present in normal nasal secretions and that the levels of IL-8 are further increased after RV infection. They also demonstrate that RVs are potent stimulators of IL-8 production and that this induction is mediated, at least in part, by an NF-kappaB-dependent transcriptional activation pathway. IL-8 may contribute to the pathogenesis of RV infection, and NF-kappaB activation may be a central event in RV-induced pathologies. PMID- 9357856 TI - Regulation of cGMP by soluble and particulate guanylyl cyclases in cultured human airway smooth muscle. AB - Although guanosine 3',5'-cyclic monophosphate (cGMP) acts as a relaxant second messenger, the regulation of intracellular cGMP has not been comprehensively studied in human airway smooth muscle. We studied the production of cGMP by cultured human airway smooth muscle cells (HASMC) after stimulation with activators of soluble guanylyl cyclase [sodium nitroprusside (SNP) and S-nitroso N-acetylpenicillamine (SNAP)] and particulate guanylyl cyclase [atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), and Escherichia coli heat stable enterotoxin (STa)]. cGMP was measured by enzyme-linked immunosorbent assay. Both SNP (10(-6) to 10(-3) M) and SNAP (10(-6) to 10(-3) M) caused concentration-dependent elevation of cGMP in the presence of the nonselective phosphodiesterase (PDE) inhibitor 3-isobutyl-1 methylxanthine (10(-3) M), with cGMP increasing 6- and 15-fold in response to SNP and SNAP, respectively, at the highest concentration tested (10(-3) M). The increases in cGMP in response to SNP (5 x 10(-5) M) and SNAP (10(-5) M) were inhibited by hemoglobin (Hb; 5 x 10(-5) M), a nitric oxide scavenger, and methylene blue (MB; 5 x 10(-4) M), an inhibitor of guanylyl cyclase. cGMP accumulation after SNAP was abolished by both Hb and MB. The response to SNP was inhibited by 79% with Hb and was abolished with MB. ANP, BNP, and CNP (10(-9) to 10(-5) M) + phosphoramidon (10(-6) M) caused a concentration-dependent elevation in cGMP with an order of potency ANP > BNP > CNP. cGMP formation in the presence of the highest concentration of the most potent natriuretic peptide (10(-5) M ANP) was two- to threefold greater than with the highest concentration of SNAP. The increase in cGMP seen with natriuretic peptides was similar in the presence or absence of phosphoramidon, a neutral endopeptidase (NEP) inhibitor, suggesting that NEP is not playing a role in modulating the effect of natriuretic peptides in HASMC. STa (400 IU/ml) had no effect on cGMP levels. SNAP- and ANP-induced cGMP accumulation was increased by the selective type V PDE inhibitors SKF-96231 and zaprinast, suggesting that type V PDE is responsible for cGMP breakdown in HASMC. These results suggest that cultured HASMC contain both soluble and particulate guanylyl cyclases. The order of potency of the natriuretic peptides ANP > BNP > CNP suggests that type A particulate membrane-bound guanylate cyclase predominates. PMID- 9357859 TI - Macrophages primed by overnight culture demonstrate a marked stimulation of surfactant protein A degradation. AB - The current study examined whether long-term culture of macrophages affects their metabolism of surfactant components. Compared with freshly isolated resting macrophages in culture for 1 h, macrophages attached to plastic dishes for 24 h showed evidence of conversion to a "primed" state with 1) an altered morphology characterized by a larger size, ruffled membranes, lamellipodia, and a "foamy" appearance after attachment to glass and 2) a fivefold greater respiratory burst in response to phorbol 12-myristate 13-acetate stimulation. On incubation with iodinated surfactant protein A (SP-A), the 24-h alveolar or tissue macrophages showed a 5- or a 23-fold greater increase in SP-A degradation, respectively, than macrophages cultured for 1 h. Conditioned media experiments demonstrated that the elevated rate of SP-A degradation after prolonged culture was not a result of proteases secreted by the macrophages. Incubation of cells with NH4Cl reduced the degradation of SP-A to a similar extent (to 33% of control values) in resting and primed tissue macrophages. On the other hand, length of time of cell culture did not affect macrophage uptake and degradation of [3H]dipalmitoylphosphatidylcholine in mixed unilamellar liposomes. Thus freshly isolated resting tissue and alveolar macrophages can be primed to specifically increase their rate of SP-A degradation. Activation of macrophages associated with lung disease may be important for SP-A metabolism and surfactant function. PMID- 9357858 TI - Dexamethasone upregulates the Na-K-ATPase in rat alveolar epithelial cells. AB - Previous studies in kidney, heart, and liver cells have demonstrated that dexamethasone regulates the expression of Na-K-ATPase. In the lungs, Na-K-ATPase has been reported in alveolar epithelial type II (ATII) cells and is thought to participate in active Na+ transport and lung edema clearance. The aim of this study was to determine whether Na-K-ATPase would be regulated by dexamethasone in cultured rat ATII cells. Regulation of the Na-K-ATPase by dexamethasone could lead to a greater understanding of its role in active Na+ transport and lung edema clearance. Rat ATII cells were isolated, plated for 24 h, and exposed to 10(-7) and 10(-8) M dexamethasone. These cells were harvested at 0, 3, 6, 12, and 24 h after dexamethasone exposure for determination of steady-state Na-K-ATPase mRNA transcript levels, protein expression, and function. The steady-state Na-K ATPase beta1-mRNA transcript levels increased in ATII cells 6, 12, and 24 h after dexamethasone exposure (P < 0.05). However, the steady-state alpha1-mRNA transcript levels were unchanged. The protein expression for the alpha1- and beta1-subunits increased in ATII cells exposed to dexamethasone compared with controls in association with a temporal increase in Na-K-ATPase function after dexamethasone exposure. These results suggest that dexamethasone regulates Na-K ATPase in ATII cells possibly by transcriptional, translational, and posttranslational mechanisms. PMID- 9357860 TI - Biosynthesis of sulfated extracellular matrices by alveolar type II cells increases with time in culture. AB - The aim of this study was to determine the extent to which sulfate incorporated into biosynthesized basement membrane (BM) components increased as isolated type II cells progress toward a more type I cell-like phenotype from 7 to 21 days in culture. Specific sulfate cytochemistry, using high iron diamine, showed that type I-like cells in 21-day cultures deposited a more highly sulfated extracellular matrix. Biosynthetic labeling experiments using [35S]cysteine or [35S]sulfate as precursors confirmed the increased capacity of 21-day type I-like cells to biosynthesize sulfated BM components compared with type II-like cells in 7-day cultures, including a novel sulfated laminin. These biochemical changes in sulfation of BM components coincide with the established phenotypic transition from type II to type I cells during prolonged culture. More importantly, the data suggest that regulation of sulfation constitutes a potential mechanism by which type I and type II cells alter their environment in such a manner as to stabilize phenotype and modulate responses to growth factors. PMID- 9357861 TI - Hypoxia stimulates human preproendothelin-1 promoter activity in transgenic mice. AB - Significant elevations in endothelin (ET)-1 levels accompany many diseases, but the underlying regulatory mechanisms are unclear. To investigate the in vivo regulation of human preproendothelin-1 (PPET-1), we examined the activity of the PPET-1 promoter in transgenic mice exposed to hypoxia. Mice expressing one of three PPET-1 promoter-luciferase (PPET-1/LUC) reporter transgenes (approximately 2.5 kb, 138 bp, or none of the 5'-flanking sequences of the PPET-1 gene) were generated. LUC expression was reduced in mice with a truncated 138-bp PPET-1 promoter. Exposure of mice bearing the 2.5-kb PPET-1/LUC transgene to hypoxia (10% O2 for 24 h) increased LUC expression sixfold in pulmonary tissue but only twofold in other tissues. In situ hybridization revealed the strongest transgene expression in the pulmonary vasculature and bronchiolar epithelium. These data are consistent with the hypothesis that hypoxic induction of the PPET-1 gene leads to increased pulmonary production of ET-1 in diseases associated with low O2 tension. PMID- 9357862 TI - Overexpression of metallothionein decreases sensitivity of pulmonary endothelial cells to oxidant injury. AB - Metallothionein (MT) is a low-molecular-weight cysteine-rich protein with extensive metal binding capacity and potential nonenzymatic antioxidant activity. Despite the sensitivity of vascular endothelium to either heavy metal toxicity or oxidative stress, little is known regarding the role of MT in endothelial cells. Accordingly, we determined the sensitivity of cultured sheep pulmonary artery endothelial cells (SPAEC) that overexpressed MT to tert-butyl hydroperoxide (t BOOH), hyperoxia, or 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN; peroxyl radical generator). Nontoxic doses of 10 microM Cd increased MT levels from 0.21 +/- 0.03 to 2.07 +/- 0.24 microg/mg and resulted in resistance to t-BOOH and hyperoxia as determined by reduction of Alamar blue or [3H]serotonin transport, respectively. SPAEC stably transfected with plasmids containing either mouse or human cDNA for MT were resistant to both t-BOOH and hyperoxia. In addition, we examined transition metal-independent, noncytotoxic AMVN-induced lipid peroxidation after metabolic incorporation of the oxidant-sensitive fluorescent fatty acid cis-parinaric acid into phospholipids and high-performance liquid chromatography separation. SPAEC that overexpressed MT after gene transfer completely inhibited peroxyl oxidation of phosphatidylserine, phosphatidylcholine, and sphingomyelin (but not phosphatidylethanolamine) noted in wild-type SPAEC. These data show for the first time that MT can 1) protect pulmonary artery endothelium against a diverse array of prooxidant stimuli and 2) directly intercept peroxyl radicals in a metal-independent fashion, thereby preventing lipid peroxidation in intact cells. PMID- 9357863 TI - Divergent regulation of 92-kDa gelatinase and TIMP-1 by HBECs in response to IL 1beta and TNF-alpha. AB - In this study, we addressed the question of whether human bronchial epithelial cells (HBECs) contribute to the regulation of 92-kDa gelatinase activity by secreting tissue inhibitor of metalloproteinase (TIMP)-1. We investigated expression of 92-kDa gelatinase and TIMP-1 in response to lipopolysaccharide (LPS) and to the proinflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha. Confluent HBECs from explants were cultured in plastic dishes coated with type I and III collagen. We demonstrated that TIMP-1 was expressed at both the protein and mRNA levels by primary cultures of HBECs. Gelatin zymography of HBEC-conditioned media showed that exposure of HBECs to LPS, IL-1beta, or TNF-alpha induced a twofold increase in the latent form of 92 kDa gelatinase production, as well as its activation. Also, quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR) demonstrated a twofold increase in the 92-kDa mRNA level in response to both cytokines. In contrast, TIMP-1 production evaluated by immunoblotting was unchanged in the presence of LPS and IL-1beta and was clearly decreased in the presence of TNF-alpha. Quantitative RT-PCR demonstrated that TIMP-1 mRNA levels remained unchanged in response to LPS or IL-1beta but decreased by 70% in the presence of TNF-alpha. All of these results strongly suggest that the control mechanisms regulating the expression of 92-kDa gelatinase and TIMP-1 by HBECs in response to inflammatory stimuli are divergent and result in an imbalance between 92-kDa gelatinase and TIMP-1 in favor of the metalloproteinase. Such an imbalance may contribute significantly to acute airway inflammation. PMID- 9357864 TI - Pulmonary dysfunction in neonatal SP-B-deficient mice. AB - Pulmonary function was assessed in newborn wild-type and homozygous and heterozygous surfactant protein B (SP-B)-deficient mice after birth. SP-B +/+ and SP-B+/- mice became well oxygenated and survived postnatally. Although lung compliance was decreased slightly in the SP-B+/- mice, lung volumes and compliances were decreased markedly in homozygous SP-B-/- mice. They died rapidly after birth, failing to inflate their lungs or oxygenate. SP-B proprotein was absent in the SP-B-/- mice and was reduced in the SP-B+/- mice, as assessed by Western analysis. Surfactant protein A, surfactant proprotein C, surfactant protein D, and surfactant phospholipid content in lungs from SP-B+/- and SP-B-/- mice were not altered. Lung saturated phosphatidylcholine and precursor incorporation into saturated phosphatidylcholine were not influenced by SP-B genotype. Intratracheal administration of perfluorocarbon resulted in lung expansion, oxygenation, and prolonged survival of SP-B-/- mice and in reduced lung compliance in SP-B+/+ and SP-B+/- mice. Lack of SP-B caused respiratory failure at birth, and decreased SP-B protein was associated with reduced lung compliance. These findings demonstrate the critical role of SP-B in perinatal adaptation to air breathing. PMID- 9357865 TI - Contribution of type I NOS to expired gas NO and bronchial responsiveness in mice. AB - Nitric oxide (NO) can be measured in the expired gas of humans and animals, but the source of expired NO (F(E)NO) and the functional contribution of the various known isoforms of NO synthase (NOS) to the NO measured in the expired air is not known. F(E)NO was measured in the expired air of mice during mechanical ventilation via a tracheal cannula. F(E)NO was significantly higher in wild-type B6SV129J +/+ mice than in mice with a targeted deletion of type I (neural) NOS (nNOS, -/-) (6.3 +/- 0.9 vs. 3.9 +/- 0.4 parts/billion, P = 0.0345, for +/+ and /- mice, respectively), indicating that approximately 40% of the NO in expired air in B6SV129 mice is derived from nNOS. Airway responsiveness to methacholine (MCh), assessed by the log of the effective dose of MCh for a doubling of pulmonary resistance from baseline (ED(200)R(L)), was significantly lower in the /- nNOS mice than in the wild-type mice (logED(200)R(L), 2.24 +/- 0.07 vs. 2.51 +/- 0.06 microg/kg, respectively; P = 0.003). These findings indicate that nNOS significantly contributes to baseline F(E)NO and promotes airway hyperresponsiveness in the mouse. PMID- 9357866 TI - rhs-TM prevents ET-induced increase in pulmonary vascular permeability through protein C activation. AB - We have previously demonstrated that recombinant human soluble (rhs) thrombomodulin (TM) inhibits the endotoxin (ET)-induced increase in pulmonary vascular permeability by inhibiting leukocyte activation. In the present study, we examined whether rhs-TM could inhibit the ET-induced increase in pulmonary vascular permeability in rats by activating protein C. rhs-TM did not inhibit ET induced increases in pulmonary vascular permeability when its protein C activation ability was selectively inhibited by a monoclonal antibody (MAb) against rhs-TM (MAb R5G12). Histological examination revealed that neutrophil infiltration in lung tissues after ET administration was significantly reduced by rhs-TM, but infiltration was not reduced by MAb R5G12-pretreated rhs-TM. ET induced intravascular coagulation was prevented by rhs-TM and by MAb R5G12 pretreated rhs-TM. However, ET-induced coagulation was not prevented by rhs-TM that had been treated with MAb F2H5, which cannot bind thrombin or activate protein C. These observations strongly suggest that rhs-TM prevents ET-induced pulmonary vascular injury by inhibiting pulmonary accumulation of leukocytes through thrombin binding and the subsequent protein C activation and may prevent ET-induced intravascular coagulation through thrombin binding. PMID- 9357867 TI - Airway surface fluid composition in the rat determined by capillary electrophoresis. AB - The apical surface of respiratory epithelial cells is covered by a thin layer of low-viscosity fluid termed airway surface fluid (ASF), about which relatively little is known. We collected samples of ASF from anesthetized rats, which were then analyzed using capillary electrophoresis, a method that enables extremely small quantities of fluid to be analyzed. We found values for Na+ (40.57 +/- 3.08 mM), K+ (1.74 +/- 0.36 mM), and Cl- (45.16 +/- 1.81 mM), indicating that this fluid is hypotonic compared with rat plasma. In contrast, the concentrations of nitrite and nitrate within ASF were higher than reported plasma values. Additionally, intravenous administration of the cholinergic agonist methacholine (MCh) resulted in a dose-dependent increase in the concentration of Na+ and Cl- within the ASF. This increase is approximately 50% in these ions after a dose of 100 ng MCh/g body wt. This animal model, together with this microanalytical technique, may be useful for investigating the in vivo regulation of ASF composition. PMID- 9357869 TI - A commentary on anesthesia gas delivery equipment and adverse outcomes. PMID- 9357868 TI - Cloning of guinea pig surfactant protein A defines a distinct cellular distribution pattern within the lung. AB - A full-length cDNA to guinea pig pulmonary surfactant protein (SP) A was cloned by screening a newborn guinea pig lung cDNA library with a human SP-A cDNA probe. The full-length guinea pig SP-A cDNA consists of 1,839 bp and is highly conserved at both nucleotide and amino acid sequence levels with those from other species. As expected, guinea pig SP-A mRNA is abundantly expressed in adolescent lung tissue and is undetectable in nonpulmonary tissues. In situ hybridization studies clearly show a unique cellular distribution pattern of SP-A mRNA within the guinea pig lung. SP-A mRNA expression is confined to cells of the alveolar epithelium with no expression in the bronchiolar epithelial cells, whereas SP-B mRNA is expressed in both alveolar and bronchiolar epithelial cell populations. This distinct expression pattern suggests that the guinea pig lung will be a useful model in which to study expression of transcription factors implicated in the regulation of SP genes. PMID- 9357870 TI - Drugs, memory, and sedation: specificity of effects. PMID- 9357871 TI - Methylnaltrexone: reversing the gastrointestinal effects of opioids. PMID- 9357872 TI - Transient neurologic symptoms: now, with phenylephrine? PMID- 9357873 TI - Richard J. Traystman: a personal story. PMID- 9357874 TI - Adverse anesthetic outcomes arising from gas delivery equipment: a closed claims analysis. AB - BACKGROUND: Anesthesia gas delivery equipment is a potentially important source of patient injury. To better define the contribution of gas delivery equipment to professional liability in anesthesia, the authors conducted an in-depth analysis of cases from the database of the American Society of Anesthesiologists Closed Claims Project. METHODS: The database of the Closed Claims Project is composed of closed US malpractice claims that have been collected in a standardized manner. All claims resulting from the use of gas delivery equipment were reviewed for recurrent patterns of injury. RESULTS: Gas delivery equipment was associated with 72 (2%) of 3,791 claims in the database. Death and permanent brain damage accounted for almost all adverse outcomes (n = 55, 76%). Equipment misuse was defined as fault or human error associated with the preparation, maintenance, or deployment of a medical device. Equipment failure was defined as unexpected malfunction of a medical device, despite routine maintenance and previous uneventful use. Misuse of equipment (n = 54, 75%) was three times more common than equipment failure (n = 17, 24%). Misconnects and disconnects of the breathing circuit made the largest contribution to injury (n = 25, 35%). Reviewers judged that 38 of 72 claims (53%) could have been prevented by pulse oximetry, capnography, or a combination of these two monitors. Overall, 56 of 72 gas delivery claims (78%) were deemed preventable with the use or better use of monitors. The year of occurrence for claims involving gas delivery equipment ranged from 1962 to 1991 and did not differ significantly from claims involving other adverse respiratory events. CONCLUSIONS: Claims associated with gas delivery equipment are infrequent but severe and continue to occur in the 1990s. Educational and preventive strategies that focus on equipment misuse and breathing circuit configuration may have the greatest potential for enhancing the safety of anesthesia gas delivery equipment. PMID- 9357875 TI - The comparative amnestic effects of midazolam, propofol, thiopental, and fentanyl at equisedative concentrations. AB - BACKGROUND: The authors evaluated the effects of midazolam, propofol, thiopental, and fentanyl on volunteer participants' memory for words and pictures at equisedative concentrations. METHODS: Sixty-seven healthy volunteers were randomized to receive intravenous infusions of midazolam (n = 11), propofol (n = 11), thiopental (n = 10), fentanyl with ondansetron pretreatment (n = 11), ondansetron alone (n = 8), or placebo (n = 16) in a double-blind design. Three increasing and then two decreasing sedative concentrations were achieved by computer-controlled infusion in each volunteer. Measures of sedation, memory, and drug concentration were obtained at each target concentration. Drug concentrations were normalized to equisedative effects using both Emax and logistic regression methods of pharmacodynamic modeling. The serum concentrations at 50% memory effect (Cp50s) were determined using four different memory end points. The relative potencies compared with midazolam for memory impairment were determined. RESULTS: Equisedative concentrations were midazolam, 64.5 +/- 9.4 ng/ml; propofol, 0.7 +/- 0.2 microg/ml; thiopental, 2.9 /- 1.0 microg/ml; and fentanyl, 0.9 +/- 0.2 ng/ml. The Cp50s for 50% loss of memory for words were midazolam, 56 +/- 4 ng/ml; propofol, 0.62 +/- 0.04 microg/ml; thiopental, 4.5 +/- 0.3 microg/ml; and fentanyl, 3.2 +/- 0.4 ng/ml. Compared with midazolam, relative potencies (with 95% confidence intervals) were propofol, 0.96 (0.44-1.78); thiopental, 0.76 (0.52-0.94); and fentanyl, 0.34 (0.05-0.76). Large effects on memory were only produced by propofol and midazolam. CONCLUSIONS: At equal sedation, propofol produces the same degree of memory impairment as midazolam. Thiopental has mild memory effects whereas fentanyl has none. Ondansetron alone has no sedative or amnesic effects. PMID- 9357877 TI - The addition of phenylephrine contributes to the development of transient neurologic symptoms after spinal anesthesia with 0.5% tetracaine. AB - BACKGROUND: Recent reports indicate that transient neurologic symptoms commonly occur after single-injection spinal anesthesia with lidocaine. Information regarding tetracaine has been limited to a single case report. In addition, little is known about the cause of these symptoms or the cofactors that affect their occurrence. The present study sought to determine whether the presence of phenylephrine or the concentration of glucose in the anesthetic solution affects the incidence of transient neurologic symptoms after spinal anesthesia with 0.5% tetracaine. METHODS: One-hundred sixty patients classified as American Society of Anesthesiologists physical status I or II who were scheduled for elective surgery on a lower limb or perineum were sequentially assigned to one of four equal groups to receive intrathecal 0.5% tetracaine in 7.5% or 0.75% glucose, with or without 0.125% phenylephrine. Patients were evaluated on postoperative day one for the presence of pain, dysesthesia, or both in the legs or buttocks by an investigator unaware of the drug given. RESULTS: Symptoms were present in 10 patients (12.5%) receiving a spinal anesthetic containing phenylephrine, but in only one patient (1.3%) receiving spinal anesthesia without phenylephrine. There was no significant difference in the incidence of symptoms between groups receiving 7.5% glucose and those receiving 0.75% glucose (8.8% and 5% of patients, respectively). CONCLUSIONS: These results suggest that adding phenylephrine to tetracaine for spinal anesthesia increases the potential for transient neurologic symptoms, but that the concentration of glucose does not affect their occurrence. PMID- 9357876 TI - Opioid-induced delay in gastric emptying: a peripheral mechanism in humans. AB - BACKGROUND: Opioids delay gastric emptying, which in turn may increase the risk of vomiting and pulmonary aspiration. Naloxone reverses this opiate action on gastric emptying, but it is not known whether this effect in humans is mediated by central or peripheral opiate antagonism. The importance of peripheral opioid receptor antagonism in modulating opioid-induced delay in gastric emptying was evaluated using methylnaltrexone, a quaternary derivative of the opiate antagonist naltrexone, which does not cross the blood-brain barrier. METHODS: In a randomized, double-blind, crossover placebo-controlled study, 11 healthy volunteers were given either placebo (saline), 0.09 mg/kg morphine, or 0.09 mg/kg morphine plus 0.3 mg/kg methylnaltrexone on three separate occasions before ingesting 500 ml deionized water. The rate of gastric emptying was measured by two methods: a noninvasive epigastric bioimpedance technique and the acetaminophen absorption test. RESULTS: The epigastric bioimpedance technique was sufficiently sensitive to detect opioid-induced changes in the rate of gastric emptying. The mean +/- SD time taken for the gastric volume to decrease to 50% (t0.5) after placebo was 5.5 +/- 2.1 min. Morphine prolonged gastric emptying to (t0.5) of 21 +/- 9.0 min (P < 0.03). Methylnaltrexone given concomitantly with morphine reversed the morphine-induced delay in gastric emptying to a t0.5 of 7.4 +/- 3.0 (P < 0.04). Maximum concentrations and area under the concentration curve from 0 to 90 min of serum acetaminophen concentrations after morphine were significantly different from placebo and morphine administered concomitantly with methylnaltrexone (P < 0.05). No difference in maximum concentration or area under the concentration curve from 0 to 90 min was noted between placebo and methylnaltrexone coadministered with morphine. CONCLUSIONS: The attenuation of morphine-induced delay in gastric emptying by methylnaltrexone suggests that the opioid effect is mediated outside the central nervous system. Methylnaltrexone may have the potential to decrease the side effects of opioid medications, which are mediated peripherally, while maintaining the central analgesia effect of the opioid. PMID- 9357878 TI - Determination of plasma concentrations of propofol associated with 50% reduction in postoperative nausea. AB - BACKGROUND: Subhypnotic doses of propofol possess direct antiemetic properties. The authors sought to determine the plasma concentration of propofol needed to effectively manage postoperative nausea and vomiting. METHODS: Patients aged 18 70 yr who were classified as American Society of Anesthesiologists physical status 1 or 2 and had surgery during general anesthesia were approached for the study. Only patients who had nausea (verbal rating score > 5 on a 0- to 10-point scale), retching, or vomiting in the postanesthetic care unit participated. Propofol was administered to these patients to achieve target plasma concentrations of 100, 200, 400, and 800 ng/ml using a computer-assisted continuous infusion device. Target concentrations were increased every 15 min until patients described at least a 50% reduction in symptoms on the verbal rating score. An arterial blood sample was obtained at each step. The measured plasma propofol concentrations were used to analyze data. Blood pressure, heart and respiratory rates, arterial blood saturation, sedation score, and overall satisfaction with treatment were recorded. RESULTS: Of the 89 patients who consented to the study, 15 patients met entry criteria and were enrolled. Five of these patients also had retching or vomiting when they entered the study. Fourteen patients responded successfully to treatment. One patient did not achieve the required response at plasma concentrations of 830 ng/ml. Hence the success rate for the treatment of postoperative nausea and vomiting was 93%. Among patients who responded, the median plasma concentration associated with an antiemetic response was 343 ng/ml. There was no difference in sedation scores from baseline and no episodes of desaturation. Hemodynamic parameters were stable during the study. CONCLUSIONS: Propofol is generally efficacious in treating postoperative nausea and vomiting at plasma concentrations that do not produce increased sedation. Simulations indicate that to achieve antiemetic plasma propofol concentrations of 343 ng/ml, a bolus dose of 10 mg followed by an infusion of approximately 10 microg x kg(-1) x min(-1) are necessary. PMID- 9357879 TI - Epidural epinephrine and clonidine: segmental analgesia and effects on different pain modalities. AB - BACKGROUND: It is not known whether epidural epinephrine has an analgesic effect per se. The segmental distribution of clonidine epidural analgesia and its effects on temporal summation and different types of noxious stimuli are unknown. The aim of this study was to clarify these issues. METHODS: Fifteen healthy volunteers received epidurally (L2-L3 or L3-L4) 20 ml of either epinephrine, 100 microg, in saline; clonidine, 8 microg/kg, in saline; or saline, 0.9%, alone, on three different days in a randomized, double-blind, cross-over fashion. Pain rating after electrical stimulation, pinprick, and cold perception were recorded on the dermatomes S1, L4, L1, T9, T6, T1, and forehead. Pressure pain tolerance threshold was recorded at S1, T6, and ear. Pain thresholds to single and repeated (temporal summation) electrical stimulation of the sural nerve were determined. RESULTS: Epinephrine significantly reduced sensitivity to pinprick at L1-L4-S1. Clonidine significantly decreased pain rating after electrical stimulation at L1 L4 and sensitivity to pinprick and cold at L1-L4-S1, increased pressure pain tolerance threshold at S1, and increased thresholds after single and repeated stimulation of the sural nerve. CONCLUSIONS: Epidural epinephrine and clonidine produce segmental hypoalgesia. Clonidine bolus should be administered at a spinal level corresponding to the painful area. Clonidine inhibits temporal summation elicited by repeated electrical stimulation and may therefore attenuate spinal cord hyperexcitability. PMID- 9357880 TI - Comparative hemodynamic depression of sevoflurane versus halothane in infants: an echocardiographic study. AB - BACKGROUND: The cardiovascular side effects of volatile anesthetics are one of the chief causes of postoperative complications in children, and infants seem to be at the greatest risk for this. This study compared cardiovascular changes at equipotent concentrations of sevoflurane and halothane in infants. METHODS: Thirty infants classified as American Society of Anesthesiologists physical status I or II who required elective surgery were randomized to receive either halothane or sevoflurane for inhalation induction. Cardiovascular and echocardiographic data were recorded in both groups at baseline and at end-tidal concentrations of 1 and 1.5 minimum alveolar concentration (MAC). RESULTS: Sevoflurane did not alter heart rate or cardiac index at all concentrations compared with awake values. Sevoflurane significantly decreased blood pressure and systemic vascular resistance compared with awake values at all concentrations. Shortening fraction and rate-corrected velocity of circumferential fiber shortening decreased at 1.5 but not at 1 MAC. Myocardial contractility assessed by stress-velocity index and stress-shortening index decreased significantly at all concentrations, but did not fall into the abnormal range at any concentration. Halothane caused a greater decrease in heart rate, shortening fraction, stress-shortening index, velocity of circumferential fiber shortening, stress-velocity index, and cardiac index at all concentrations than did sevoflurane. CONCLUSION: Sevoflurane causes a lesser decrease in cardiac output than does halothane in infants. PMID- 9357881 TI - Effects of rapid increases of desflurane and sevoflurane to concentrations of 1.5 MAC on systemic vascular resistance and catecholamine response during cardiopulmonary bypass. AB - BACKGROUND: Airway irritation was hypothesized to trigger the transient cardiovascular stimulation associated with desflurane. The authors administered desflurane during cardiopulmonary bypass (CPB), thus avoiding airway contact, and compared the effects of rapid increases of desflurane to 1.5 MAC on systemic vascular resistance index (SVRI) and catecholamine response to those of 1.5 MAC sevoflurane. METHODS: Forty-eight patients, undergoing elective coronary bypass surgery, were randomly allocated to receive either desflurane or sevoflurane during hypothermic (32-33 degrees C) nonpulsatile CPB at exhaust gas concentrations of 1.5 MAC for 15 min. SVRI was calculated at baseline, 1, 2, 3, 4, 5, 7, 9, 12, and 15 min after starting volatile anesthetics' delivery. Plasma catecholamine concentrations were determined in 12 desflurane-treated patients and 12 sevoflurane-treated patients at baseline, 5, and 15 min. RESULTS: The time course of deltaSVRI, (changes in SVRI from baseline), from baseline to 5 min was significantly different between desflurane- and sevoflurane-treated patients, whereas there was no difference from 7 to 15 min. In the desflurane group, SVRI from 1 to 7 min remained unchanged to baseline level, thereafter declining to significantly lower values at 9, 12, and 15 min compared with values from 0 to 5 min, whereas sevoflurane produced an immediate and significant reduction in SVRI. With desflurane, catecholamine concentrations remained unchanged to baseline level at 5 and 15 min; with sevoflurane, they decreased with time. CONCLUSIONS: The authors' results indicate that desflurane is associated with a different hemodynamic and catecholamine response compared with sevoflurane when administered into the oxygenator's gas supply line during CPB. PMID- 9357883 TI - Measurement of cardiac output by pulse dye densitometry using indocyanine green: a comparison with the thermodilution method. AB - BACKGROUND: A new method for determining cardiac output (CO, l/min) using dye dilution combined with pulse dye densitometry (PDD), based on the principle of pulse oximetry, has been developed. The aim of the study was to determine the accuracy and precision of PDD by comparing it with the thermodilution method. METHODS: A prospective study was performed in 22 patients having surgery who were monitored using a pulmonary arterial catheter. In addition to the catheter, a specially designed photodetector was placed on the nasal wing. Ten milliliters of ice-cold indocyanine green dissolved in a 5% glucose solution (0.5 mg/ml) was injected. The dye and thermal dilution curves were simultaneously measured to calculate CO. Three to six injections were performed before and after surgery. Paired data were assessed in absolute terms, and the percentage errors were calculated by the degree of agreement and compared at three levels of CO (low < or = 3.5 < medium < or = 6 < high) by analysis of variance. RESULTS: The mean and SDs of the differences between dye and thermodilution CO were 0.16 +/- 0.80 l/min or 4.5 +/- 19.6% for 191 paired data. Measurement after surgery failed in one patient. The percentage error with low CO (9.3 +/- 19.3%) was greater (P < 0.05) than those obtained with other CO. CONCLUSIONS: Pulse dye densitometry could measure CO repeatedly in patients having major surgery with the same degree of accuracy as the thermodilution method; however, a considerable degree of error was observed in some patients. PMID- 9357882 TI - Bispectral index monitoring allows faster emergence and improved recovery from propofol, alfentanil, and nitrous oxide anesthesia. BIS Utility Study Group. AB - BACKGROUND: The bispectral index (BIS), a parameter derived from the electroencephalograph (EEG), has been shown to correlate with increasing sedation and loss of consciousness. This study determined whether addition of BIS monitoring to standard anesthetic practice results in improvements in the conduct of anesthesia or in patient outcomes. METHODS: Three hundred two patients receiving a propofol-alfentanil-nitrous oxide anesthetic were studied at four institutions. Thirty-four patients were initially enrolled to determine preexisting anesthetic practice and patient outcomes at each institution. Subsequent patients were randomized to either standard clinical practice (SP group), or standard practice plus BIS monitoring (BIS group). In all patients, the anesthesiologist attempted to provide a stable anesthetic with the fastest possible recovery. BIS was recorded for all patients, but viewed only in the BIS group. In the BIS group, propofol infusions were adjusted to achieve a target BIS between 45-60, increasing to 60-75 during the final 15 min of the case. In the SP group, propofol dose adjustments were made based only on standard clinical signs. Drug use, intraoperative responses, and patient recovery parameters were recorded. RESULTS: Demographics were similar between groups. Compared with the SP group, patients in the BIS group required lower normalized propofol infusion rates (134 vs. 116 microg x kg[-1] x min[-1]; P < 0.001), were extubated sooner (11.22 vs. 7.25 min; P < 0.003), had a higher percentage of patients oriented on arrival to PACU (43% vs. 23%; P < 0.02), had better postanesthesia care unit (PACU) nursing assessments (P < 0.001), and became eligible for discharge sooner (37.77 vs. 31.70 min; P <0.04). There was no significant difference in the incidence of intraoperative responses between the groups. CONCLUSIONS: Titrating propofol with BIS monitoring during balanced anesthesia decreased propofol use and significantly improved recovery. Intraoperative course was not changed. These findings indicate that the use of BIS may be valuable in guiding the administration of propofol intraoperatively. PMID- 9357884 TI - Factors influencing the tracheal fluctuation of inhaled nitric oxide in patients with acute lung injury. AB - BACKGROUND: Inhaled nitric oxide (NO) improves arterial oxygenation in patients with acute lung injury (ALI) by selectively dilating pulmonary vessels perfusing ventilated lung areas. It can be hypothesized that NO uptake from the lung decreases with increasing ventilation perfusion mismatch. This study was undertaken to determine the factors influencing the fluctuation of tracheal NO concentration over the respiratory cycle as an index of NO pulmonary uptake in patients with ALI. METHODS: By using a prototype system (Opti-NO) delivering a constant flow of NO only during the inspiratory phase, 3 and 6 ppm of NO were administered during controlled mechanical ventilation into a lung model and to 11 patients with ALI. All patients had a thoracic computed tomography (CT) scan. Based on an analysis of tomographic densities, lungs were divided into three zones: normally aerated (-1.000 to -500 Hounsfield units [HU]), poorly aerated ( 500 to -100 HU), and nonaerated (-100 to +100 HU), and the volume of each zone was computed. Concentrations of NO in the inspiratory limb and trachea were continuously measured by a fast-response chemiluminescence apparatus. RESULTS: In the lung model, tracheal NO concentration was stable with minor fluctuation. In contrast, in patients, tracheal NO concentration fluctuated widely during the respiratory cycle (55 +/- 10%). Because uptake of NO from the lungs was absent in the lung model but present in the patients, this fluctuation was considered as an index of pulmonary uptake of NO. This was further substantiated by (1) the coincidence of the peak and minimum tracheal NO concentration with the end inspiratory and end-expiratory phases, respectively, and (2) continued decrease of tracheal NO concentration during prolonged expiratory phase. In patients with ALI, the fluctuation of tracheal NO concentration expressed as the difference between inspiratory and expiratory NO concentrations divided by inspiratory NO concentration was greater at 6 ppm than at 3 ppm (P < 0.01), was linearly correlated with normally aerated lung volume, inversely correlated with alveolar dead space and with poorly aerated lung volume. CONCLUSION: In patients with ALI, fluctuation of tracheal NO concentration over the respiratory cycle can be considered as an index of NO uptake from the lungs that depends on aerated lung volume and perfusion of ventilated lung areas. At bedside, it may be used to follow the evolution of ventilation-perfusion mismatch. PMID- 9357886 TI - Titration of volatile anesthetics using bispectral index facilitates recovery after ambulatory anesthesia. AB - BACKGROUND: The bispectral (BIS) index has previously been shown to be a quantifiable measure of the sedative and hypnotic effects of anesthetic drugs. This study was designed to assess the effect of BIS monitoring on the utilization of volatile anesthetics and their recovery profiles after ambulatory surgery. METHODS: Sixty consenting women undergoing outpatient laparoscopic tubal ligation procedures were randomly assigned to one of four treatment groups. After a standardized induction, anesthesia was maintained with either desflurane (Groups I and II) or sevoflurane (Groups III and IV) in combination with nitrous oxide, 65%, and fentanyl. In the control groups (Groups I and III), the anesthesiologists were blinded to the BIS value, and the volatile anesthetics were administered according to standard clinical practice. In Groups II and IV, the volatile anesthetics were titrated to maintain the BIS value at 60. The volatile anesthetic usage and the times from discontinuation of anesthesia to verbal response, orientation, and home-readiness were recorded. RESULTS: During the maintenance period, the BIS values were significantly lower in the control groups (mean, 42) compared with the BIS-titrated groups (mean, 60). The volatile anesthetic usage in the BIS-titrated groups was 30-38% lower (P < 0.05) compared with the control groups. Similarly, the times to verbal responsiveness were 30 55% shorter in the BIS-titrated (vs. control) groups. CONCLUSIONS: Titrating desflurane and sevoflurane using the BIS monitor decreased their utilization and contributed to a faster emergence from anesthesia in outpatients undergoing laparoscopic tubal ligation procedures. PMID- 9357885 TI - Dexmedetomidine does not alter the sweating threshold, but comparably and linearly decreases the vasoconstriction and shivering thresholds. AB - BACKGROUND: Clonidine decreases the vasoconstriction and shivering thresholds. It thus seems likely that the alpha2 agonist dexmedetomidine will also impair control of body temperature. Accordingly, the authors evaluated the dose dependent effects of dexmedetomidine on the sweating, vasoconstriction, and shivering thresholds. They also measured the effects of dexmedetomidine on heart rate, blood pressures, and plasma catecholamine concentrations. METHODS: Nine male volunteers participated in this randomized, double-blind, cross-over protocol. The study drug was administered by computer-controlled infusion, targeting plasma dexmedetomidine concentrations of 0.0, 0.3, and 0.6 ng/ml. Each day, skin and core temperatures were increased to provoke sweating and then subsequently reduced to elicit vasoconstriction and shivering. Core-temperature thresholds were computed using established linear cutaneous contributions to control of sweating, vasoconstriction, and shivering. The dose-dependent effects of dexmedetomidine on thermoregulatory response thresholds were then determined using linear regression. Heart rate, arterial blood pressures, and plasma catecholamine concentrations were determined at baseline and at each threshold. RESULTS: Neither dexmedetomidine concentration increased the sweating threshold from control values. In contrast, dexmedetomidine administration reduced the vasoconstriction threshold by 1.61 +/- 0.80 degrees C x ng(-1) x ml (mean +/- SD) and the shivering threshold by 2.40 +/- 0.90 degrees C x ng(-1) x ml. Hemodynamic responses and catecholamine concentrations were reduced from baseline values, but they did not differ at the two tested dexmedetomidine doses. CONCLUSIONS: Dexmedetomidine markedly increased the range of temperatures not triggering thermoregulatory defenses. The drug is thus likely to promote hypothermia in a typical hospital environment; it is also likely to prove an effective treatment for shivering. PMID- 9357888 TI - Predictive factors in global and anesthesia satisfaction in ambulatory surgical patients. AB - BACKGROUND: Patient satisfaction is one of the variables that affect the outcome of health care and the use of health-care services. As more procedures are performed on an ambulatory basis, the role of the anesthesiologist becomes more important. To improve the delivery of care, the predictors of dissatisfaction with the entire process (global dissatisfaction) of ambulatory surgery and with anesthesia itself must be identified. The authors conducted a hypothesis generating study to identify predictors; specifically, they hypothesized that satisfaction with anesthesia was a predictor of global satisfaction with ambulatory surgery and that 24-h postoperative symptoms were a predictor of satisfaction with anesthesia. METHODS: The authors prospectively studied 5,228 consecutive patients having surgery in the ambulatory setting during a 1-yr period. Preoperative, intraoperative, and postoperative variables were gathered and patient satisfaction was assessed using a postoperative telephone questionnaire administered 24 h after operation in 2,730 respondents. Significant univariate variables and clinically important variables were entered into multiple logistic regression models. Qualitative data on dissatisfaction were obtained by asking patients' reasons for dissatisfaction. RESULTS: Sixty-eight of the 2,730 respondents (2.5%) had global dissatisfaction with ambulatory surgery. Nine of these patients were dissatisfied with anesthesia. Dissatisfaction with anesthesia was associated with a 12-fold increase in global dissatisfaction (P = 0.0001). Thirty-one of the 2,730 respondents (1.1%) were dissatisfied with anesthesia. An increasing number of symptoms occurring 24 h after operation was associated with an exp(0.28 x N)-fold increase in dissatisfaction with anesthesia for N number of symptoms (P = 0.0001). Qualitative data showed that the most common reason for global dissatisfaction with ambulatory surgery was personal preference for inpatient care (26%), whereas intraoperative and postoperative adverse outcomes were the major causes of dissatisfaction with anesthesia (88%). CONCLUSIONS: Dissatisfaction with anesthesia is a predictor of global dissatisfaction with ambulatory surgery. An increasing number of symptoms 24 h after operation is a predictor of dissatisfaction with anesthesia. The rate of global dissatisfaction and anesthesia dissatisfaction is very low. The predictors from this model need to be validated by a second data set from either this or another center. Given the low rate of dissatisfaction, a focused study testing specific interventions to improve patient satisfaction would be difficult. PMID- 9357887 TI - Alfentanil blocks reflex pupillary dilation in response to noxious stimulation but does not diminish the light reflex. AB - BACKGROUND: Estimation of the mu-agonist opioid effect in anesthetized and paralyzed patients is often imprecise and can be obscured by concomitant administration of drugs that affect the sympathetic nervous system, such as beta adrenergic blocking agents. As an alternative to hemodynamic measures of opioid effect, the authors tested the hypothesis that the pupillary light reflex or pupillary reflex dilation correlated with alfentanil concentrations during isoflurane anesthesia. METHODS: Six volunteers were anesthetized on 4 days with 0.8% isoflurane. Alfentanil was administered intravenously to target total plasma concentrations of 0, 25, 50, and 100 ng/ml. A 5-s tetanic electrical stimulus was applied to the skin. Pupil size and the pupillary light reflex were recorded before and after alfentanil administration, and before and for 8 min after the stimulus. RESULTS: Alfentanil exponentially impaired reflex pupillary dilation, decreasing the maximum response amplitude from 5 mm at 0 ng/ml, to 2.3 mm at 25 ng/ml, to 1.0 mm at 50 ng/ml, and finally to 0.2 mm at 100 ng/ml. In contrast, only the highest concentration of alfentanil depressed the dilation of the pupil in the first 2 s after the stimulus. Alfentanil administration had no effect on the pupillary light reflex. CONCLUSIONS: Dilation of the pupil in response to a noxious stimulus is a measure of opioid effect in isoflurane-anesthetized volunteers. In contrast, the pupillary light reflex is unaffected by alfentanil during isoflurane anesthesia. These data suggest that stimulus-induced pupillary dilation may be used to evaluate the analgesic component of a combined volatile and opioid anesthetic. PMID- 9357889 TI - Development of a measure of patient satisfaction with monitored anesthesia care: the Iowa Satisfaction with Anesthesia Scale. AB - BACKGROUND: The authors describe development of the Iowa Satisfaction with Anesthesia Scale (ISAS) for monitored anesthesia care (MAC). Patients complete the self-administered written questionnaire before discharge from the hospital. The authors designed the ISAS to measure satisfaction with MAC itself, not the perioperative experience. Patients respond to eleven statements (e.g., "I felt pain") by placing a mark along a six-choice vertical response column (e.g., "Disagree moderately") below each statement. The mean of their responses to each of the 11 statements gives a single number, which is a quantitative measure of a patient's satisfaction with their MAC. METHODS: Adult, English-speaking patients completed the questionnaires following admission to a phase II postanesthesia care unit after MAC. RESULTS: Response rate for MAC was 92% (86 of 94 patients). Patients completed the questionnaire in 4.6 +/- 2.3 min. Internal consistency, Cronbach's alpha, equaled 0.80. Patients' scores were positively correlated with those predicted by their anesthesia provider (r2 = 0.23) and with responses to the question "I was satisfied with my anesthetic care" (Kendall's tau = +0.41). Scores on initial and repeat questionnaires were positively correlated (r2 = 0.74). Scores on initial questionnaires and those completed within 4.4 +/- 1.7 days postoperatively were positively correlated (r2 = 0.76). CONCLUSIONS: The authors have developed and tested an internally consistent, reliable, and valid measure of patient satisfaction with MAC. PMID- 9357890 TI - Intrathecal sufentanil for labor analgesia does not cause a sympathectomy. AB - BACKGROUND: Intrathecal sufentanil (ITS) is frequently used to provide analgesia during labor. Decreases in blood pressure and sensory changes in this circumstance suggest that ITS may have a local anesthetic effect and thus cause a sympathectomy. To determine whether ITS given to laboring women causes a sympathectomy, the authors evaluated central and lower extremity temperature changes after ITS administration. These findings were compared with those in a control group of women having spinal anesthesia with bupivacaine for elective cesarean section in whom an extensive sympathectomy was expected. METHODS: Twenty parturients classified as American Society of Anesthesiologists' physical status 1 or 2 had temperatures measured centrally, at the calf, and at the great toe at frequent intervals after receiving 10 microg ITS for labor analgesia (sufentanil group, n = 10), or hyperbaric bupivacaine 12 mg in their spinal anesthetic for cesarean section (bupivacaine group, n = 10). Calf-to-toe temperature indices (C T) were calculated by subtracting toe temperature from calf temperature. A decrease in this index means that the toe had warmed compared with the calf and is an indication of vasodilation and a sympathectomy. RESULTS: There was no significant change in the C-T indices or central temperature in the sufentanil group, but the C-T indices and central temperature decreased significantly in the bupivacaine group. CONCLUSIONS: The decreases in the C-T index and central temperature in the bupivacaine group indicate the presence of a sympathectomy. The lack of change in the C-T indices and central temperature in the sufentanil group indicates that no significant vasodilation occurred. Therefore, the decrease in blood pressure seen after ITS administration for labor analgesia is unlikely to be the result of a sympathectomy. PMID- 9357891 TI - Estrogen-induced changes in protein binding of bupivacaine during in vitro fertilization. AB - BACKGROUND: Patients having in vitro fertilization (IVF) procedures that use gonadotropin-releasing hormone agonist down-regulation undergo hormonal manipulation of estrogen concentrations to induce oocyte maturation. After achieving minimal estrogen concentrations (baseline), stimulation increases estrogen concentrations to supraphysiologic levels, leading to egg retrieval. The isolated effect of estrogen on protein binding has not previously been reported. This study was conducted to measure the effect of estrogen concentrations on protein binding of two concentrations of bupivacaine, 1 microg/ml and 5 microg/ml, corresponding, respectively, to systemic concentrations expected after administration of epidural anesthesia and associated with bupivacaine toxicity. Serum proteins were measured to address the mechanism. METHODS: Twenty-nine healthy women undergoing IVF procedures were enrolled and venous samples were drawn at times of minimal and maximal estrogen concentrations. The percentage of free bupivacaine was determined at fixed concentrations of 1 and 5 microg/ml. Serum albumin and alpha1-acid glycoprotein concentrations were measured at baseline and at retrieval in a group of 24 women. RESULTS: The percentage of free bupivacaine increased between times of minimal and maximal serum estrogen concentrations, corresponding to decreased protein binding. Concentrations of serum albumin and alpha1-acid glycoprotein decreased between baseline and retrieval times. CONCLUSIONS: Patients undergoing IVF procedures demonstrate a decrease in protein binding of bupivacaine from baseline concentrations. These changes may be explained by a decrease in albumin and alpha1-acid glycoprotein. During anesthesia for egg retrieval, clinicians should consider the implications of increased free fraction of drug, especially for highly protein-bound agents. PMID- 9357892 TI - Computer simulation of the effects of alterations in blood flows and body composition on thiopental pharmacokinetics in humans. AB - BACKGROUND: Understanding the influence of physiological variables on thiopental pharmacokinetics would enhance the scientific basis for the clinical usage of this anesthetic. METHODS: A physiological pharmacokinetic model for thiopental previously developed in rats was scaled to humans by substituting human values for tissue blood flows, tissue masses, and elimination clearance in place of respective rat values. The model was validated with published serum concentration data from 64 subjects. The model was simulated after intravenous thiopental administration, 250 mg, over 1 min, to predict arterial plasma concentrations under conditions of different cardiac outputs, degrees of obesity, gender, or age. RESULTS: The human pharmacokinetic model is characterized by a steady state volume of distribution of 2.2 l/kg, an elimination clearance of 0.22 l/min, and a terminal half-life of 9 h. Measured thiopental concentrations are predicted with an accuracy of 6 +/- 37% (SD). Greater peak arterial concentrations are predicted in subjects with a low versus a high cardiac output (3.1 and 9.4 l/min), and in subjects who are lean versus obese (56 and 135 kg). Acutely, obesity influences concentrations because it affects cardiac output. Prolonged changes are due to differences in fat mass. Changes with gender and age are relatively minor. CONCLUSIONS: The physiological pharmacokinetic model developed in rats predicts thiopental pharmacokinetics in humans. Differences in basal cardiac output may explain much of the variability in early thiopental disposition between subjects. PMID- 9357893 TI - Intravenous anesthetics attenuate phenylephrine-induced calcium oscillations in individual pulmonary artery smooth muscle cells. AB - BACKGROUND: The authors investigated the effects of intravenous anesthetics on alpha-adrenergic-induced oscillations in intracellular free calcium concentration ([Ca2+]i) in individual pulmonary artery smooth muscle cells (PASMCs). METHODS: PASMCs were cultured from explants of canine intrapulmonary artery. Fura-2-loaded PASMCs were continuously superfused with phenylephrine (10 microM) at 37 degrees C on the stage of an inverted fluorescence microscope. Measurement of [Ca2+]i was via a dual wavelength spectrofluorometer. Intravenous anesthetics were added to the superfusate to assess their effects on the phenylephrine-induced [Ca2+]i oscillations. RESULTS: Resting [Ca2+]i was 103 +/- 6 nM. Phenylephrine stimulated [Ca2+]i oscillations, reaching a peak concentration of 632 +/- 20 nM and a frequency of 1.53 +/- 0.14 transients/min. The effects of phenylephrine were dose dependent. The effects of intravenous anesthetics on phenylephrine-induced [Ca2+]i oscillations were dose-dependent. Ketamine (100 microM) reduced the amplitude (221 +/- 22 nM) but not the frequency (1.48 +/- 0.11/min) of the oscillations, whereas thiopental (100 microM) decreased the amplitude (270 +/- 20 nM) and the frequency (1.04 +/- 0.10/min). Propofol (100 microM) and the Intralipid vehicle inhibited the amplitude (274 +/- 11 nM) but not the frequency (1.39 +/- 0.11/min) of the oscillations. The effects of ketamine and thiopental, but not propofol, were evident at clinically relevant concentrations. CONCLUSION: Ketamine, thiopental, and propofol exerted differential effects to inhibit the amplitude or the frequency of phenylephrine-induced [Ca2+]i oscillations in individual PASMCs. Thus, intravenous anesthetics may alter the pulmonary vascular response to alpha-adrenoreceptor activation by directly inhibiting [Ca2+]i signaling in PASMCs. PMID- 9357894 TI - Halothane and xanthine oxidase increase hepatocellular enzyme release and circulating lactate after ischemia-reperfusion in rabbits. AB - BACKGROUND: Multiple-organ injury often occurs after aortic occlusion reperfusion. Oxidants derived from xanthine oxidase have been implicated as a source of injury after aortic occlusion-reperfusion. Halogenated anesthetics modify oxidant-mediated injury. The current study determined if halothane modifies hepatocellular enzyme release (e.g., alanine aminotransferase) and circulating lactate after aortic occlusion-reperfusion. METHODS: Rabbits were randomly assigned to one of four groups that underwent 40 min of thoracic aortic occlusion and 2 h of reperfusion: Two groups were given either halothane or fentanyl plus droperidol anesthesia and two groups were given either anesthetic and sodium tungstate (xanthine oxidase inactivator). Each of the four groups was then matched with a similarly treated group that did not undergo aortic occlusion. RESULTS: Halothane anesthesia was associated with significantly (P < 0.05) increased release of alanine aminotransferase (34 +/- 9 U/l at baseline and 539 +/- 370 U/l at 120 min of reperfusion; mean +/- SD) and increased plasma lactate concentrations (2.8 +/- 2.0 mM at baseline and 12.1 +/- 9.7 mM at 120 min of reperfusion) after aortic occlusion-reperfusion compared with fentanyl plus droperidol anesthesia (alanine aminotransferase, 33 +/- 12 U/l and 148 +/- 109 U/l; lactate, 3.4 +/- 2.0 mM and 3.8 +/- 1.2 mM at baseline and 120 min of reperfusion, respectively). Inactivation of xanthine oxidase significantly decreased the release of hepatocellular enzymes (P < 0.05) and decreased circulating lactate in animals anesthetized with halothane after aortic occlusion reperfusion. CONCLUSIONS: Halothane increased hepatocellular enzyme release and circulating lactate after aortic occlusion-reperfusion compared with fentanyl plus droperidol anesthesia. Xanthine oxidase activity inactivation also decreased hepatocellular enzyme activity release during reperfusion. These findings justify further investigations to determine if halogenated anesthetics modify tissue injury in clinical settings involving oxidant stress. PMID- 9357895 TI - Transient neuronal depolarization induces tolerance to subsequent forebrain ischemia in rats. AB - BACKGROUND: Minor cortical injury has previously been shown to improve survival in animals subjected to ischemic insults. Although the mechanism by which an ischemia-tolerant state is achieved is not clear, transient neuronal depolarization is thought to play a central role in the development of the tolerance. One way of producing transient neuronal depolarization is by the induction of cortical spreading depression (CSD). The present study was conducted to evaluate the effect of preischemic transient depolarization, induced by CSD, on postischemic neuronal outcome in rats. METHODS: Unilateral CSD was induced by application of KCl to the frontal cortex (CSD hemisphere) in three groups of isoflurane-anesthetized rats (CSD groups; n = 8/group). Sham animals (n = 12) did not undergo CSD. In a fifth group (n = 8), ketamine was administered during KCl application to inhibit CSD. One, three, or seven days after CSD, animals were subjected to forebrain ischemia produced by bilateral carotid artery occlusion. Injury to the striatum, hippocampus, and cortex was evaluated in hematoxylin and eosin-stained brain sections 3 days after ischemia. RESULTS: Preischemic CSD reduced postischemic injury in the ipsilateral cortex. The ratio of the number of injured neurons in the CSD hemisphere to that in the non-CSD hemisphere was significantly less in the groups subjected to CSD 1 day (0.51 +/- 0.33), 3 days (0.56 /- 0.22), and 7 days (0.40 +/- 0.17) before ischemia than in the sham operated group (1.11 +/- 0.47). In the ketamine group (CSD inhibition), there were no differences in the extent of injury in the two hemispheres (ratio = 0.84 +/- 0.47). Injury to the striatum and hippocampus was similar among the groups. Within each group, injury to these subcortical structures in the CSD hemisphere was not different from that in the non-CSD hemisphere. CONCLUSIONS: The data suggest that preischemic depolarization induced by CSD results in an adaptive response that reduces the vulnerability of cortical neurons to subsequent ischemic injury (ischemic tolerance). PMID- 9357896 TI - Bupivacaine inhibition of L-type calcium current in ventricular cardiomyocytes of hamster. AB - BACKGROUND: The local anesthetic bupivacaine is cardiotoxic when accidentally injected into the circulation. Such cardiotoxicity might involve an inhibition of cardiac L-type Ca2+ current (ICa,L). This study was designed to define the mechanism of bupivacaine inhibition of ICa,L. METHODS: Cardiomyocytes were enzymatically dispersed from hamster ventricles. Certain voltage- and time dependencies of ICa,L were recorded using the whole-cell patch clamp method in the presence and absence of different concentrations of bupivacaine. RESULTS: Bupivacaine, in a concentration-dependent manner (10-300 microM), tonically inhibited the peak amplitude of ICa,L. The inhibition was characterized by an increase in the time of recovery from inactivation and a negative-voltage shift of the steady-state inactivation curve. The inhibition was shown to be voltage dependent, and the peak amplitude of ICa,L could not be restored to control levels by a wash from bupivacaine. CONCLUSIONS: The inhibition of ICa,L appears, in part, to result from bupivacaine predisposing L-type Ca channels to the inactivated state. Data from washout suggest that there may be two mechanisms of inhibition at work. Bupivacaine may bind with low affinity to the Ca channel and also affect an unidentified metabolic component that modulates Ca channel function. PMID- 9357897 TI - Effect of midazolam on propofol-induced positive affective state assessed by place conditioning in rats. AB - BACKGROUND: The effect of either midazolam or the combination of midazolam and propofol on the affective state was assessed in rats at subanesthetic doses and at recovery from anesthesia. METHODS: The putative drug(s)-induced affective states were repeatedly paired with one of two distinguishable compartments of an experimental cage, whereas the vehicle(s)-induced effect was repeatedly paired with the other compartment. During a subsequent choice test for one compartment over the other, the rats' preference for the drug(s)-paired compartment over the vehicle(s)-paired compartment is indicative of a pleasant state induced by the drug(s). In experiment 1, rats were conditioned with different doses of midazolam either at subanesthetic states or at recovery from anesthesia. In experiment 2, groups of rats were conditioned with different combinations of midazolam and propofol either at subanesthetic states or at recovery from anesthesia induced jointly by midazolam (10 mg/kg) and propofol (60 mg/kg). Experiment 3 was conducted in the same way as experiment 2, except that midazolam was paired with both compartments. In addition, these groups were tested not only in an undrugged state but also in a drugged (with midazolam) state. RESULTS: In experiment 1, rats exhibited a place preference for the environment previously associated with midazolam, at subanesthetic and anesthetic doses. Experiment 2 showed that a propofol-induced place preference was found to be dose-dependently suppressed by midazolam. Experiment 3 replicated the findings of experiment 2 and extended them to the mechanism by which midazolam blocked a propofol-induced place preference. CONCLUSIONS: Midazolam administered before propofol blocked the expression of a propofol-induced pleasant state. PMID- 9357898 TI - The effect of thiopental and propofol on NMDA- and AMPA-mediated glutamate excitotoxicity. AB - BACKGROUND: Glutamate excitotoxicity has been implicated as an important cause of ischemic, anoxic, epileptic, and traumatic neuronal damage. Glutamate receptor antagonists have been shown to reduce anoxic, ischemic, and epileptic damage. The effects of thiopental and propofol on N-methyl-D-aspartate (NMDA) and alpha-amino 3-hydroxy-5-methyl-4-isoxazole proprionate (AMPA)-induced neuronal damage were investigated in this study. METHODS: The Schaffer collateral pathway was stimulated, and a postsynaptic-evoked population spike was recorded from the CA1 pyramidal cell layer of rat hippocampal slices. The recovery of the population spike amplitude was an indicator of neuronal viability. The duration of NMDA (25 microM) or AMPA (15 or 10 microM) treatment was 10 min. Thiopental (600 microM), propofol (112 microM), or the vehicle was present 15 min before, during, and 10 min after the NMDA or AMPA treatment. RESULTS: Thiopental prolonged the time required to completely block the population spike after the addition of NMDA or AMPA. Thiopental improved the recovery of the population spike after 25 microM NMDA (79% vs. 44%) and 15 microM AMPA (50% vs. 15%). Propofol worsened the recovery of the population spike from NMDA-induced damage. The recovery was 8% with propofol compared with 40% with NMDA alone. Propofol did not significantly alter the AMPA-induced neuronal damage. CONCLUSIONS: Thiopental attenuates NMDA- and AMPA-mediated glutamate excitotoxicity. This may be one way barbiturates reduce anoxic, ischemic, and epileptic damage. Propofol enhances NMDA-induced neuronal damage. These results demonstrate that thiopental and propofol have different properties with respect to glutamate excitotoxicity. PMID- 9357899 TI - Isoflurane and halothane do not alter the enhanced afterload sensitivity of left ventricular relaxation in dogs with pacing-induced cardiomyopathy. AB - BACKGROUND: The afterload dependence of left ventricular (LV) relaxation is accentuated in the failing heart. The authors tested the hypothesis that isoflurane and halothane alter the afterload sensitivity of LV relaxation in dogs with pacing-induced cardiomyopathy. METHODS: Dogs (n = 6) were chronically instrumented for measurement of LV and aortic pressures and subendocardial segment length. Hemodynamics were recorded, and LV relaxation was evaluated with a time constant of isovolumic relaxation (tau) under control conditions and during decreases and increases in LV load produced by abrupt inferior vena caval (IVC) occlusion and phenylephrine (intravenous infusion), respectively, in the conscious state and during isoflurane and halothane anesthesia (1.5 MAC) on separate days before and after the development of pacing-induced cardiomyopathy. The slope (R) of the tau versus LV end-systolic pressure (P[es]) relation was also used to determine the afterload sensitivity of LV relaxation. RESULTS: IVC occlusion and phenylephrine produced similar or less profound changes in P(es), regional end-systolic force (an index of LV afterload), and end-systolic segment length in cardiomyopathic compared with healthy dogs. However, IVC occlusion and phenylephrine caused more pronounced alterations in tau in conscious and isoflurane- and halothane-anesthetized dogs after the development of cardiomyopathy. R was also greater in cardiomyopathic compared with healthy dogs (e.g., 0.32 +/- 0.03 before pacing to 1.00 +/- 0.13 ms/mmHg in conscious dogs). No differences in the load dependence of LV relaxation were observed between the conscious and anesthetized states before and after production of LV dysfunction. CONCLUSIONS: The results indicate that isoflurane and halothane do not alter the afterload dependence of LV relaxation in the normal and cardiomyopathic heart. The lack of effect of the volatile anesthetics is probably related to anesthetic induced reductions in the resistance to LV ejection concomitant with simultaneous negative inotropic effects. PMID- 9357901 TI - Informed consent. PMID- 9357900 TI - The effect of imidazoline receptors and alpha2-adrenoceptors on the anesthetic requirement (MAC) for halothane in rats. AB - BACKGROUND: Recent evidences have documented that several pharmacologic actions of alpha2-adrenoceptor agonists are mediated via activation of not only alpha2 adrenoceptors, but also by imidazoline receptors, which are nonadrenergic receptors in the central nervous system. However, the effect of imidazoline receptors on the anesthesia is not well known, and it is important to clarify the effects of both receptors on anesthesia. METHODS: Seventy-two rats were anesthetized with halothane, and the anesthetic requirement for halothane was evaluated as minimum alveolar concentration (MAC). The MAC for halothane was determined in the presence of dexmedetomidine (0, 10, 20, and 30 microg/kg, intraperitoneally [IP]), a selective alpha2-adrenoceptor agonist with weak affinity for imidazoline receptors. Then, the authors evaluated the inhibitory effect of rauwolscine (20 mg/kg, IP), an alpha2-adrenoceptor antagonist with little affinity for imidazoline receptors, on the MAC-reducing action of dexmedetomidine (30 microg/kg). Further, the effect of rilmenidine (20, 50, 100, 1000 microg/kg, IP), a selective imidazoline receptor agonist, on the MAC for halothane was also investigated. RESULTS: Dexmedetomidine decreased the MAC for halothane dose-dependently, and this MAC-reducing action of dexmedetomidine was completely blocked by rauwolscine. Rilmenidine alone did not change the MAC for halothane. CONCLUSIONS: The present data indicate that the anesthetic sparing action of dexmedetomidine is most likely mediated through alpha2- adrenoceptors, and the stimulation of imidazoline receptors exerts little effect on the anesthetic requirement for halothane. PMID- 9357902 TI - Critical hemoglobin desaturation will occur before return to an unparalyzed state following 1 mg/kg intravenous succinylcholine. PMID- 9357903 TI - Spinal hematoma following spinal anesthesia in a patient with spina bifida occulta. PMID- 9357905 TI - Inhaled nitric oxide in sickle cell disease with acute chest syndrome. PMID- 9357904 TI - Extreme hemodilution due to massive blood loss in tumor surgery. PMID- 9357907 TI - Management of massive grain aspiration. PMID- 9357906 TI - Factor XII deficiency and cardiopulmonary bypass: use of a novel modification of the activated clotting time to monitor anticoagulation. PMID- 9357908 TI - Respiratory failure after laparoscopic cholecystectomy in a patient with chronic hemidiaphragm paralysis. PMID- 9357909 TI - Intraoperative immediate diagnosis of acute obstruction of tricuspid valve and pulmonary embolism due to renal cell carcinoma with transesophageal echocardiography. PMID- 9357910 TI - Fiberoptic intracranial pressure monitoring during magnetic resonance imaging. PMID- 9357911 TI - Lactic acidosis as a serious perioperative complication of antidiabetic biguanide medication with metformin. PMID- 9357912 TI - Severe dysphonia after the use of a laryngeal mask airway. PMID- 9357913 TI - Anesthesia-related deaths during obstetric delivery in the United States. 1979 1990. PMID- 9357914 TI - Cardiac arrest and epidural anesthesia. PMID- 9357915 TI - Mechanism of hyperchloremic nonanion gap acidosis. PMID- 9357916 TI - Dilutional acidosis: a nonentity? PMID- 9357917 TI - Dilutional acidosis or altered strong ion difference. PMID- 9357918 TI - Decrease in the total amount of extracellular bicarbonate is not dilution. PMID- 9357919 TI - CACI in cardiac surgery. PMID- 9357921 TI - Postdural puncture headache and epidural blood patch. PMID- 9357920 TI - Platelet aggregation inhibited by sevoflurane, or by ethanol? PMID- 9357922 TI - Severe bradycardia after remifentanil. PMID- 9357923 TI - Workshop on how to perform clinical outcome studies. University of Munster, Munster, Germany, May 22-23, 1997. PMID- 9357924 TI - Immunologic basis of chronic allergic diseases: clinical messages from the laboratory bench. AB - During the past decade there have been significant advances in our understanding of the mechanisms underlying allergic responses. Immediate hypersensitivity reactions are mediated primarily by mast cells in an IgE-dependent manner. After the local release of various mediators, proinflammatory cytokines, and chemokines, there is a cell-mediated response that is dominated by eosinophils and T lymphocytes. The majority of T cells in early allergic reactions are memory T cells secreting helper type 2 (TH2)-like cytokines, i.e. IL-4, IL-5, and IL-13, but not interferon-gamma. These cytokines regulate IgE synthesis and promote eosinophil differentiation and cell survival, thus contributing to allergic inflammatory responses. Failure to control immune activation early in the course of allergic inflammation may blunt the response to glucocorticoid therapy and contribute to long-term morbidity of disease. The identification of key cells and cytokines involved in the initiation and maintenance of allergic inflammation is likely to become an important therapeutic target in the future management of this important group of diseases. PMID- 9357925 TI - Medium chain 3-ketoacyl-coenzyme A thiolase deficiency: a new disorder of mitochondrial fatty acid beta-oxidation. AB - A Japanese male neonate died at 13 d of age after presenting at 2 d of age with vomiting, dehydration, metabolic acidosis, liver dysfunction, and terminal rhabdomyolysis with myoglobinuria. Multiple urine organic acid analyses consistently revealed a markedly elevated excretion of lactic acid, 3 hydroxybutyric acid, and saturated and unsaturated C6-C16 dicarboxylic acids, with predominant C12-C16 species. Oxidation of [1-14C]octanoic acid in cultured skin fibroblasts was significantly reduced (0.59 nmol/h/mg of protein; controls, 1.93 +/- 0.65), [1-14C]palmitic acid oxidation was 1.11 nmol/h/mg of protein (controls, 1.63 +/- 0.41). A systematic study of the catalytic activities of nine enzymes of the beta-oxidation cycle using the respective optimal substrate revealed a deficiency of a single enzyme not previously associated with a metabolic disorder, medium chain 3-ketoacyl-CoA thiolase (patient, 3.9 nmol/min/mg protein; controls (n = 6), 10.2 +/- 2.3). Immunoprecipitation with antibodies raised against medium chain 3-ketoacyl-CoA thiolase revealed a 60% decrease compared with controls. PMID- 9357926 TI - Cerebrospinal fluid and plasma total homocysteine and related metabolites in children with cystathionine beta-synthase deficiency: the effect of treatment. AB - The neurologic complications of cystathionine beta-synthase deficiency are thought to be secondary to accumulation of homocyst(e)ine in the CNS. Treatment of this disorder with betaine has been shown to improve the behavior of individuals, to reduce plasma total homocysteine, and to correct secondary abnormalities of serine. To test the hypothesis that homocyst(e)ine accumulates within the CNS and that this can be reduced by treatment with betaine, we measured total homocysteine and related metabolites in the plasma of 10 children with cystathionine beta-synthase deficiency and cerebrospinal fluid of five children before and during betaine therapy. In plasma, betaine significantly lowered total homocysteine (but not to the normal range) and had a variable effect on methionine. In the cerebrospinal fluid, total homocysteine was raised before treatment (mean 1.2 microM) and was significantly reduced by betaine (mean 0.32 microM) but not to the normal range (<0.10 microM). Cerebrospinal fluid methionine was raised before and during treatment, but betaine did not cause a significant further increase. Cerebrospinal fluid serine was significantly reduced before treatment and rose to the normal range with betaine. Cerebrospinal fluid S-adenosylmethionine was normal before treatment and rose significantly with treatment; there were no significant changes in cerebrospinal fluid 5 methyltetrahydrofolate. The demonstration of accumulation of homocysteine within the CNS lends support to the hypothesis that this may be one cause of the neurologic complications of cystathionine beta-synthase deficiency. Betaine is effective in reducing cerebrospinal fluid homocysteine, but concentrations are still significantly raised during treatment. PMID- 9357927 TI - Deficient muscle carnitine transport in primary carnitine deficiency. AB - Primary carnitine deficiency is associated with deficient blood and tissue carnitine concentrations. The clinical syndrome is dominated by heart and skeletal muscle symptoms, and the clinical response to oral carnitine supplementation is life-saving. Carnitine uptake has been shown to be defective in cultured skin fibroblasts and leukocytes obtained from patients with this condition. We report a new case of primary carnitine deficiency and offer direct evidence consistent with an impairment of carnitine uptake in differentiating muscle culture. The patient presented with severe and progressive cardiomyopathy and moderate proximal limb weakness. Plasma and muscle carnitine levels were very low, and the maximal rate of carnitine transport in cultured fibroblasts was deficient. An asymptomatic sister with intermediate levels of carnitine in plasma showed partially deficient carnitine uptake in fibroblasts, indicating heterozygosity. The patient's condition improved dramatically with oral carnitine therapy. Further studies were performed in cultured muscle cells at different stages of maturation, which demonstrated deficient maximal rates of carnitine uptake. Our findings are consistent with the concept that primary carnitine deficiency is the result of a generalized defect involving carnitine transport across tissue membranes. PMID- 9357928 TI - Angiotensin-converting enzyme inhibition decreases growth factor expression in the neonatal rat kidney. AB - The renin-angiotensin system plays an important role in renal growth and development: exposure of the fetus or neonate to angiotensin-converting enzyme (ACE) inhibitors increases mortality and results in growth retardation and abnormal renal development. This study was designed to investigate the effects of ACE inhibition in the neonatal rat on the expression of genes known to modulate renal cellular proliferation, cell interactions, and extracellular matrix. Newborn rat pups were treated with enalapril (30 mg/kg/d) or vehicle for 14 d, and kidneys were removed for Northern analysis of mRNA for transforming growth factor-beta1 (TGF-beta1), prepro epidermal growth factor (EGF), clusterin, and renin. Distribution of TGF-beta1, EGF, and clusterin was also determined by immunohistochemistry. Enalapril treatment resulted in 40% mortality by d 14, reduced body and kidney weight, decreased glomerular area, and caused tubular dilatation (p < 0.05 versus vehicle group). Enalapril decreased renal TGF-beta1 and EGF mRNA expression, and increased renal clusterin and renin expression (p < 0.05). Renal tubular immunoreactive EGF was decreased, and clusterin was increased by enalapril treatment. These results indicate that ACE inhibition in the developing kidney reduces the renal expression of critical growth factors, which may account for renal growth impairment. Clusterin expression may increase either due to blockade of tonic angiotensin-mediated inhibition, or as an adaptive response to renal ischemia. PMID- 9357929 TI - Increased leptin messenger RNA and serum leptin levels in children with Prader Willi syndrome and nonsyndromal obesity. AB - To study the potential role of the ob gene pathway in childhood obesity, we have investigated leptin mRNA levels in s.c. adipose tissue obtained from nonobese prepubertal children (n = 20), obese nonsyndromal children (n = 6), and children with Prader-Willi syndrome (n = 6) by in situ hybridization histochemistry. We have also investigated the fasting serum leptin levels in such children. Compared with nonobese children, leptin mRNA expression was higher both in children with Prader-Willi syndrome and in children with nonsyndromal obesity (p < 0.01). Furthermore, the serum leptin levels were also significantly higher in both children with Prader-Willi syndrome and nonsyndromal obesity compared with the nonobese children (p < 0.001). However, no significant differences in adipose tissue leptin mRNA or serum leptin levels were observed between children with Prader-Willi syndrome and nonsyndromal obese children. As expected both fasting serum leptin levels and leptin mRNA expression levels correlated to body mass index (rs = 0.80 and 0.73, respectively, p < 0.005). No difference in leptin expression between Prader-Willi syndrome and nonsyndromal childhood obesity could be revealed in the present study. However, differences in the hypothalamic response to leptin between the two forms of obesity cannot be excluded. PMID- 9357930 TI - Platelet-activating factor (PAF) up-regulates plasma and tissue PAF acetylhydrolase activity in the rat: effect of cycloheximide. AB - Platelet-activating factor (PAF) is a proinflammatory phospholipid mediator implicated in necrotizing enterocolitis. Regulation of PAF-acetylhydrolase (AH), the enzyme degrading PAF, is poorly understood. In this study we found that administration of a dose of PAF (1.5 microg/kg, i.v.), which does not cause gross intestinal injury, increased plasma and intestinal PAF-AH in the rat. Cycloheximide (CHX, 5 mg/kg, i.v.) reduced the activity of plasma (but not intestinal tissue) AH in control, as well as in PAF-injected rats, and aggravated systemic inflammation and tissue injury in the latter. The intestinal necrosis induced by PAF and CHX was ameliorated by posttreatment with WEB2170 (a PAF antagonist), indicating a role of endogenous PAF in mediating injury. Both WEB2170 and anti-TNF antibody reduced PAF-induced AH activity in intestinal tissue, but not in the plasma. Allopurinol largely prevented the injury induced by PAF and CHX, but had no effect on the up-regulation of AH. We conclude: 1) de novo protein synthesis is required to maintain physiologic AH level in the plasma; 2) PAF up-regulates plasma and intestinal AH activity; 3) CHX enhances the injurious effect of PAF; 4) endogenous PAF and TNF also play a role in the up regulation of intestinal AH; the former probably mediating the intestinal injury by PAF; and 5) reactive oxygen species may mediate the injurious effect of PAF plus CHX, but do not contribute to the regulation of AH by PAF. PMID- 9357931 TI - Thyroid function in very preterm infants: influences of gestational age and disease. AB - It is not known how immaturity and disease influence postnatal thyroid function in infants <30 wk of gestational age. We performed serial measurements of plasma thyroxine (T4), free T4 (FT4), triiodothyronine (T3), reverse T3 (rT3), TSH, and T4-binding globulin (TBG) in 100 infants of <30 wk of gestation, during the first 8 postnatal weeks, to investigate the influences of disease and gestational age on the time course of thyroid hormones. One hundred infants were divided twice into two groups: 1) in a group of 25-28 and of 28-30 wk of gestation; and 2) in a sick and a healthy group, with similar gestational ages. The time course of T4, FT4, T3, TSH, and TBG, but not rT3 differed significantly (p < 0.005) between the gestational age groups. T4 and FT4 decreased to levels below the cord blood value with a deeper FT4 nadir on d 7 in the youngest group. Disease decreased T4, FT4, T3, TSH, and TBG concentrations especially during the 1st wk after birth (p < 0.005). However, the FT4 nadir on d 7 was similar in sick and healthy infants. After 3 wk, T4, FT4, T3, and TBG were higher in the sick group compared with the healthy group. rT3 levels were not increased in sick infants. We conclude that the extent of the FT4 decrease after birth in infants of <30 wk gestation is mainly influenced by gestational age and probably reflects a transient depletion of thyroidal hormone reserves. rT3 cannot be used as a marker of nonthyroidal illness in very preterm infants. PMID- 9357932 TI - Hypothyroidism delays fetal stratum corneum development in mice. AB - The epidermal permeability barrier, required for terrestrial life, is localized to lipid-enriched lamellar membranes in the extracellular spaces of the stratum corneum (SC). Immaturity of the SC is a significant contributor to morbidity and mortality in premature infants. Previous studies have shown that supraphysiologic concentrations of thyroid hormone accelerate epidermis/SC ontogenesis. Here we studied SC development in Hyt/Hyt mice who are genetically hypothyroid due to a mutation in the TSH receptor. In control mice on d 18 of gestation (term 19.5 d), only focal areas displayed a mature SC membrane pattern. By 19 d of gestation there was a mature multilayered SC with lamellar unit structures filling the extracellular spaces similar to that seen in mature mice. In Hyt/Hyt mice SC development was delayed at both 18 and 19 d of gestation. In both strains of mice, within the first day after birth there were no differences in epidermal or SC appearance, and the SC was fully mature. These findings indicate that thyroid hormone plays a physiologic role during normal intrauterine development of the SC. However, normal SC maturation ultimately occurs, indicating that thyroid hormone is not absolutely essential. Previous studies have shown that glucocorticoids accelerate SC development in euthyroid rats, and in the present study we demonstrate that glucocorticoids also accelerate SC ontogenesis in euthyroid mice. In contrast, in Hyt/Hyt mice glucocorticoids did not accelerate or normalize SC development, indicating that the glucocorticoid effect on SC maturation requires a euthyroid state or that glucocorticoids act via thyroid hormone. These studies demonstrate that thyroid hormone status is an important regulator of fetal SC development. PMID- 9357933 TI - Modulation of the effect of bilirubin on protein phosphorylation by lysine containing peptides. AB - Protein phosphorylation is an important mechanism for regulation of cell processes. Bilirubin inhibits phosphorylation of several peptides/proteins by a number of different kinases, and this may contribute to the toxic effects of bilirubin on cells, particularly neurons. Bilirubin binds to lysine residues on both albumin and ligandin. The ATP-binding subdomain II on protein kinases contains an invariant lysine, which might hypothetically be involved in mediation or modulation of bilirubin-inhibition of protein phosphorylation. We have studied the ability of lysine-containing peptides to modulate the effects of bilirubin, using phosphorylation of a phospholemman peptide catalyzed by protein kinase A as a model system. Addition of bilirubin (50 microM) decreased the activity of the catalytic subunit of protein kinase A by 75%. A synthetic lysine-containing decapeptide which mimicked part of subdomain II on the protein kinase family was partially able to prevent the bilirubin effects. Similar effects were not observed with two other decapeptides in which lysine had been replaced by arginine or alanine. Polylysine (100 microM) completely prevented the inhibitory effect of 50 microM bilirubin, whereas polyglutamate and polyarginine did not have this effect. Poly-D-lysine and poly-L-lysine appeared to be equivalent in their ability to prevent the bilirubin effect. These data support the notion that binding of bilirubin to lysine may play a role in the mediation and/or modulation of bilirubin neurotoxicity. PMID- 9357934 TI - Duodenal motor responses in preterm infants fed formula with varying concentrations and rates of infusion. AB - Feeding intolerance is frequently reflected in preterm infants by delayed gastric emptying. Gastric emptying is delayed by the physical characteristics of ingested nutrient as well as the rate of feeding. Because gastric emptying is dependent upon duodenal function, the present studies were undertaken to assess duodenal motor responses to feeding of differing nutrient content and rate of feeding. Using a Latin square design we recorded duodenal motor responses in 14 preterm infants given four test feedings in random order over 18 h. Three were given as a 120-min infusion containing no nutrient, a 10 cal/oz formula, and a 20 cal/oz formula. The fourth test feeding consisted of a 20 cal/oz formula given as a bolus over 15 min. Although caloric density was altered, osmotic load and nutrient proportions of the formulas were not. Motor responses were recorded using a low compliance continuous perfusion manometric system. When infants were fed "water" and half-strength formula as a slow infusion, they demonstrated little or no duodenal motor response to feeding. When these infants were fed full strength formula as a slow infusion, they displayed a brisk increase in motor activity (p < 0.05), but profound motor quiescence when fed the same volume by bolus over 15 min (p < 0.05). Of the four test feedings, only full-strength formula given as a slow infusion triggered adult-like duodenal motor responses to feeding. We speculate that feedings of full-strength formula given slowly by infusion will improve feeding tolerance. PMID- 9357935 TI - Treatment of extrahepatic biliary atresia with interferon-alpha in a murine infectious model. AB - The etiology of extrahepatic biliary atresia (EHBA) in newborns remains unknown, although a first infectious animal model with complete obstruction of the common bile duct could be established. Intraperitoneal inoculation of newborn Balb/c mice with rhesus rotavirus induced cholestasis, leading, in most cases, to biliary atresia with lethal outcome, similar to EHBA in human newborns. The influence of interferon-alpha (IFN-alpha) on the hepatotropism of rotavirus infection was investigated in this animal model. Single-dose therapy with 10000 IU of IFN-alpha protected all rhesus rotavirus-infected pups from cholestatic disease. The same dose, injected 5 d after infection, had no protective effect. Starting with onset of cholestatic symptoms, the treatment with 10000 IU of IFN alpha daily showed good results in 29 mice. Seventy-six percent of the mice recovered after 1 wk of therapy. Histologic investigation revealed normal findings in the hepatobiliary tract of clinically normal mice. Twenty-one percent of the descendants of infected and prophylactic IFN-alpha-treated mice showed cholestatic symptoms after infection with rhesus rotavirus (79% in an untreated control group) and a milder form of the illness. In conclusion, we found that prophylactic treatment with IFN-alpha prevented the hepatobiliary system of newborn Balb/c mice from severe damage by rhesus rotavirus in this artificially designed infectious model for EHBA. Infected and icteric mice, treated for 1 wk with IFN-alpha, had good prospects for recovery and prevention of complete and irreversible occlusion of the extrahepatic bile ducts. Infected and prophylactic IFN-alpha-treated dams gave good protection to their descendants. This means that EHBA in this model could probably be averted by maternal antibodies against rotavirus. PMID- 9357936 TI - The frequency of cells secreting interferon-gamma and interleukin-4, -5, and -10 in the blood and duodenal mucosa of children with cow's milk hypersensitivity. AB - Enzyme-linked immunoabsorbant spots (ELISPOTs) have been used to analyze the frequency of cells spontaneously secreting interferon-gamma (INF-gamma), IL-4, IL 5, or IL-10 in mononuclear cells isolated from the blood of children with cow's milk-sensitive enteropathy (CMSE), cow's milk allergy (CMA), and age-matched controls. In addition, cytokine profiles of duodenal lamina propria lymphocytes were compared in patients with CMSE and control subjects. In blood, spontaneous cytokine-secreting cells were uncommon, but there was significantly increased IFN gamma, IL-4, IL-5, and IL-10 ELISPOTs in children with CMSE and CMA compared with control subjects. IL-4 ELISPOTs were significantly greater in the blood of children with CMA compared with those with CMSE. In the lamina propria the frequencies of spontaneous cytokine-secreting cells were high compared with that in blood. Significantly increased ELISPOTs for IFN-gamma and IL-4 were found in CMSE compared with controls. IL-5 ELISPOTs were unchanged, and IL-10 ELISPOTs were reduced in CMSE compared with controls. These results show a general enhancement of Th1 and Th2-type cytokine-secreting cells in the blood of children with cow's milk hypersensitivity, although the increased IL-4-secreting cells in blood in CMA may be of relevance in view of the fact that this disease is IgE mediated. In the lamina propria, there is also enhancement of IFN-gamma- and IL-4 secreting cells in CMSE compared with control subjects; however, cells secreting IFN-gamma are 10 times more numerous than cells secreting IL-4, showing a dominance of Th1-type responses in both controls and CMSE patients. PMID- 9357937 TI - Antiinflammatory effects of human milk on chemically induced colitis in rats. AB - We examined the effects of a human milk diet on rats with chemical colitis induced with a 4% acetic acid enema. Colonic myeloperoxidase activity was used as a surrogate marker for neutrophil infiltration. Control rats fed rat chow had little colonic myeloperoxidase activity; geometric mean, 0.27 U/g of tissue. Rats with colitis fed rat chow had significantly increased colonic myeloperoxidase activity (geometric mean, 6.76 U/g, p < 0.01 versus no colitis), as did rats with colitis fed infant formula or Pedialyte (geometric mean, 6.92 and 8.13 U/g, respectively, both p < 0.01 versus no colitis). Animals with colitis fed human milk had significantly lower colonic myeloperoxidase activity (geometric mean, 2.34 U/g) than did animals with colitis fed either chow or infant formula (p < 0.001). Similar effects were seen in rats with colitis fed infant formula supplemented with recombinant human IL-1 receptor antagonist (geometric mean, 1.95 U/g). These data show that orally administered human milk has an antiinflammatory effect on chemically induced colitis in rats, which may be mediated in part by IL-1 receptor antagonist contained in human milk. PMID- 9357938 TI - Antibodies to Escherichia coli and Shigella flexneri in milk from undernourished mothers: studies on sodium dodecyl sulfate-polyacrylamide gel electrophoresis separated antigens. AB - Analysis of IgA, IgM, and IgG antibodies against Escherichia coli O6, its lipopolysaccharide (LPS), and Shigella flexneri were performed in the milk of moderately undernourished Guatemalan women receiving either a low or a high calorie supplement, using SDS-PAGE. As expected, the immunostaining analysis of milk antibodies showed that IgA was the predominant isotype in both groups. Concerning the other Igs, antibodies against O6 LPS were mainly of the IgM isotype, whereas IgG antibodies were more prominent than IgM against the bacterial whole cell preparations. Seven to nine distinct bands, ranging in molecular mass from 13.5 to 109 kD were selected for each antigen to compare the milk antibodies between the two groups of women. After a 20-wk supplementation period, the IgA and IgG antibodies to the E. coli, O6 LPS, and S. flexneri were not found negatively affected by a low calorie intake. A significantly lower immunostaining intensity was, however, obtained for the low calorie intake group regarding the IgM antibody activity against four high molecular mass bands of the E. coli whole cell preparation. A decreased immunostaining intensity was also found in the same group for IgM antibodies against two bands of E. coli O6 LPS when analyzing paired samples collected at d 0 and wk 20. No differences were found for IgM antibodies against any of the S. flexneri antigens. In conclusion, the results suggest that low calorie intake does not significantly affect the production of milk IgA antibodies to E. coli and S. flexneri antigens in these women. Still, IgM antibodies against certain proteins and LPS molecules of E. coli may be decreased. IgG antibodies, although also present in milk, seemed to be directed mainly against bacterial proteins and not to LPS. PMID- 9357939 TI - Serum amyloid A and high density lipoprotein participate in the acute phase response of Kawasaki disease. AB - In this study we report changes in HDL concentration and composition in acute and convalescent Kawasaki disease. Notable reductions in plasma HDL-cholesterol (0.54 +/- 0.2 mmol/L, normal level 0.7-1.81 mmol/L) and apolipoprotein A-I (apoA-I) (56 +/- 28 mg/dL, normal level 141 +/- 22 mg/dL) were observed in all 24 patients studied during the acute phase of Kawasaki disease. These changes were accompanied by the marked appearance of serum amyloid A (SAA) protein in the plasma, associated with HDL3-like lipoprotein particles. The distribution of apoA I was analyzed in five patients and showed a significant increase in lipid-free apoA-I in the bottom fraction (28.8 +/- 4.1%, normal range 10-15%), suggesting displacement of apoA-I from the HDL particles by SAA. Within 2 wk after acute Kawasaki disease, levels of HDL-cholesterol and apoA-I returned to the normal range, and SAA disappeared from the plasma. The HDL of patients with Kawasaki disease was markedly enriched in triglyceride even in the absence of changes in total plasma triglyceride. The core composition of HDL returned to the normal range more slowly than the plasma HDL-cholesterol and apoA-I levels. This suggests that Kawasaki disease has a profound effect on the lipoprotein profile acutely and a more subtle sustained effect on the HDL composition. We interpret these changes as manifestations of the acute phase response in Kawasaki disease. PMID- 9357940 TI - Apoptosis of CD4+ and CD8+ T cells isolated immediately ex vivo correlates with disease severity in human immunodeficiency virus type 1 infection. AB - Apoptosis of CD4+ and CD8+ T cells has been shown in peripheral blood mononuclear cells (PBMCs) from HIV-infected adults analyzed after overnight culture. Because cell death may be an artifact of in vitro culture, and because there is little information on apoptosis in pediatric HIV disease, we undertook a cross-sectional analysis of apoptosis in PBMCs analyzed immediately ex vivo in HIV-infected children and adults. PBMCs from 22 children, four adolescents, and nine adults and seronegative age-matched control subjects were stained for CD4 and CD8 surface markers. Apoptotic cells were detected in a newly characterized flow cytometric assay by diminished forward and increased side scatter. Children with the most advanced disease had 9.9% (SEM 1.8) apoptotic CD4+ T cells above control, significantly higher than in asymptomatic patients [0.4% (SEM 2.3)], those with mild disease [2.2% (SEM 1.83)], and those with moderate disease [2.5 (SEM 3.6)] (p = 0.015). The percentages of both CD4+ and CD8+ T cell apoptosis were directly related to CD4+ T cell depletion (R2 = 0.23; p = 0.006; n = 32 and R2 = 0.2; p = 0.012; n = 30, respectively). Patients who responded to antiretroviral therapy with the greatest increase in CD4+ T cell percentage had the least CD4+ T cell apoptosis (R2 = 0.15; p = 0.1; n = 19). These findings show that the rate or extent of T cell death by apoptosis percentage of T cell apoptosis is significantly increased in HIV-infected children. The observed correlation of both CD4+ and CD8+ T cell apoptosis with CD4+ T cell depletion suggests that apoptosis plays a role in HIV pathogenesis and may be a useful marker of disease activity. PMID- 9357941 TI - Diagnostic and predictive value of auditory evoked responses in preterm infants: I. Patient characteristics and long-term neurodevelopmental outcome. AB - The diagnostic and predictive value of brainstem, middle latency, and cortical auditory evoked responses, obtained in the neonatal period, in 81 preterm infants was assessed in relation to neurodevelopmental outcome. Eighteen healthy term infants served as a control group. In this report the patient characteristics and neurodevelopmental outcome are presented. The preterm infants were neonatally classified according to risk category and gestational age. At 5 y of age the neurodevelopmental outcome was assessed based on neurologic and neuropsychologic evaluations. The neuropsychologic test results showed the highest IQ scores in term infants, intermediate IQ scores in low risk preterm infants, and lowest IQ scores in high risk preterm infants. The intermediate IQ scores in the low risk preterm group were due to significantly lower test scores in a small subgroup of low risk preterm infants. In a post hoc analysis 12 low risk preterm infants with an unfavorable outcome could be identified. The neuropsychologic test results of the remaining low risk infants showed no clear differences compared with the term infants. The results suggest that the unfavorable outcome of the low risk preterm group as a whole is due to moderate to severe impairment of the few, rather than slight impairment of the majority. PMID- 9357942 TI - Diagnostic and predictive value of auditory evoked responses in preterm infants: II. Auditory evoked responses. AB - In this study, the diagnostic and predictive value of brainstem, middle latency, and cortical auditory evoked responses (BMC-AERs) obtained in the neonatal period in 81 preterm infants was assessed in relation to neurodevelopmental outcome. The preterm infants were neonatally classified according to risk category and gestational age. The BMC-AERs were analyzed with respect to detectability, latencies, and amplitudes as well as derived latency and amplitude measures. At 5 y of age the neurodevelopmental outcome was assessed from neurologic and neuropsychologic evaluations. The results showed that BMC-AER differences mainly correlated with risk category (low risk/high risk) and to some extent with degree of prematurity. In view of these findings the degree of prematurity and the effect of risk category have to be taken into account, when BMC-AERs are applied in the preterm period to predict neurodevelopmental outcome. In this study the BMC-AERs for infants with abnormal neurodevelopmental outcome were scarcely distinguishable from the BMC-AERs for infants with normal neurodevelopmental outcome. Thus far, this and previous reports have indicated that BMC-AERs in preterm infants are useful in maturational studies and with infants showing symptoms related to lesions or dysfunction of the peripheral and/or central auditory system. For predicting neurodevelopmental outcome in preterm infants, BMC-AERs are of limited clinical value. PMID- 9357943 TI - Sensorimotor function and neuropathology five to six weeks after hypoxia-ischemia in seven-day-old rats. AB - Various therapeutic interventions after hypoxia-ischemia (HI) have been shown to reduce brain injury in the short-term perspective, but it remains uncertain whether such findings are accompanied by long-term functional and structural improvements. HI was induced in 7-d-old rats as follows. The left carotid artery was ligated, and the rat was exposed to 100 min of hypoxia (7.70% oxygen in nitrogen). At postnatal d 42 the rats were assessed using four sensorimotor tests. The results were correlated with the extent of brain damage expressed as volume of deficit of the left hemisphere as percent of the right hemisphere. In the grip-traction test, the time to falling was 2.2 times shorter in the HI animals compared with controls (p < 0.01). Asymmetries of limb-placing and foot faults (p < 0.001) were detected in HI animals, and the motor function was abnormal in the postural reflex test (p < 0.001). We found a moderate correspondence between functional and neuropathologic outcome (r = 0.842, p < 0.001). A set of four easily performed sensorimotor tests is presented for the long-term evaluation of neurologic function in the 7-d-old rat model of HI. PMID- 9357944 TI - Apoptosis in the brains of infants suffering intrauterine cerebral injury. AB - This study addressed the hypothesis that in human infants severe in utero insults induce a significant proportion of brain cells to undergo apoptosis. Morphologic criteria were used to quantify apoptosis and necrosis in the cingulate gyrus of two groups of infants: six infants who died after severe birth asphyxia with hypoxic-ischemic encephalopathy, and six others who suffered unexpected and apparently sudden intrauterine death at or close to term. The fraction of apoptotic cells was much higher than basal levels determined in animal experiments, and within both groups increased in proportion to the severity of injury as determined by total cell death (p < 0.05). The mean fraction of apoptotic cells was similar in asphyxiated infants, 8.3% (95% confidence interval for the population, 3.7-12%), and in stillbirths, 6.7% (0.2-13.6%). In the asphyxiated group, 20.8% (11-30.6%) of cells were necrotic, but significantly less necrosis, 3% (0.4-5.6%), was seen in stillborn infants (p < 0.05). Cell death was apoptotic after birth asphyxia in 26% (1-51%) and 78% (41-100%) in stillborn infants. In situ end labeling studies confirmed the presence of DNA fragmentation in apoptotic cells. These results demonstrate that infants who die after intrauterine insults, both those with evidence of delayed cerebral injury after hypoxia-ischemia and those without, have a significant number of cells in the brain with the morphologic characteristics of apoptosis. They confirm that apoptosis contributes significantly to cerebral damage in the perinatal period. PMID- 9357945 TI - Cross-spectral analysis of cerebral autoregulation dynamics in high risk preterm infants during the perinatal period. AB - In preterm infants intraventricular hemorrhage occurs predominantly within the perinatal period, which may be due to a "lost autoregulation" of cerebral blood flow (CBF). In this study, perinatal autoregulation dynamics were investigated in high risk preterm infants by cross-spectral analysis (CSA), which is a statistical tool in the analysis of time series. In 15 ventilated preterm infants of 25-32 gestational weeks, a total number of 30 records were made between 24 and 96 h of life. Doppler-derived CBF velocity (CBFv), used as a quantitative measure for CBF, and direct mean arterial blood pressure (MABP) were measured continuously for 10 min. The spectral power of low frequency (LF, 0.02-0.2 Hz) oscillations in CBFv and MABP was quantified by spectral analysis. From the results of CSA, a LF phase-shift between the CBFv and MABP LF oscillations was calculated in each record. Within the study group, the LF spectral power of CBFv and MABP was initially low and increased significantly until 96 h of life. The LF phase-shift was about 0 degrees at 24 h and increased significantly to 55 degrees at 96 h of life. The initially low LF spectral power of CBFv and MABP may indicate a perinatal depression of autonomic nervous centers, which are thought to control LF oscillations of vital parameters. In the light of a high pass filter model for autoregulation, the initially low LF phase-shift may indicate an initially impaired autoregulation, which supports the "lost autoregulation" hypothesis. PMID- 9357946 TI - Modest hypothermia preserves cerebral energy metabolism during hypoxia-ischemia and correlates with brain damage: a 31P nuclear magnetic resonance study in unanesthetized neonatal rats. AB - Recent studies have shown that mild to moderate (modest) hypothermia decreases the damage resulting from hypoxic-ischemic insult (HI) in the immature rat. To determine whether suppression of oxidative metabolism during HI is central to the mechanism of neuroprotection, 31P nuclear magnetic resonance (NMR) spectroscopy was used to measure high energy metabolites in 7-d postnatal rats under conditions of modest hypothermia during the HI. The rats underwent unilateral common carotid artery ligation followed by exposure to hypoxia in 8% oxygen for 3 h. Environmental temperature was decreased by 3 or 6 degrees C from the control temperature, 37 degrees C, which reliably produces hemispheric damage in over 90% of pups. The metabolite parameters and tissue swelling (edema) at 42 h recovery varied very significantly with the three temperatures. Tissue swelling was 26.9, 5.3, and 0.3% at 37, 34, and 31 degrees C, respectively. Core temperature and swelling were also measured, with similar results, in parallel experiments in glass jars. Multislice magnetic resonance imaging, histology, and triphenyltetrazolium chloride staining confirmed the fairly uniform damage, confined to the hemisphere ipsilateral to the ligation. The NMR metabolite levels were integrated over the last 2.0 h out of 3.0 h of HI, and were normalized to their baseline for all surviving animals (n = 25). ATP was 47.9, 69.0, and 83.0% of normal, whereas the estimator of phosphorylation potential (phosphocreatinine/inorganic phosphorus) was 16.9, 27.8, and 42.6% of normal at 37, 34, and 31 degrees C, respectively. There was a significant correlation of both phosphocreatinine/inorganic phosphorus (p < 0.0001) and ATP levels (p < 0.0001) with brain swelling. Abnormal brain swelling and thus damage can be reliably predicted from a threshold of these metabolite levels (p < 0.0001). Thus for all three temperatures, a large change in integrated high energy metabolism during HI is a prerequisite for brain damage. With a moderate hypothermia change of 6 degrees C, where there is an insufficient change in metabolites, there is no subsequent HI brain damage. In general, treatment for HI in our 7-d-old rat model should be aimed at preserving energy metabolism. PMID- 9357947 TI - Thyrotropin-releasing hormone accelerates fetal mouse lung ultrastructural maturation via stimulation of extra thyroidal pathway. AB - Maternal administration of TSH-releasing hormone (TRH) in the euthyroid mouse accelerates fetal lung ultrastructural maturation. However, the mechanism(s) of TRH in fetal lung development remains unclear; it could be due to its neuroendocrine and/or neurotransmitter effects. Although the neuroendocrine effect of TRH is mediated via stimulation of the fetal pituitary-thyroid axis, the neurotransmitter effect is mediated via stimulation of fetal autonomic nervous system activity. In the hyt/hyt mouse there is a point mutation in the beta subunit of the TSH receptor in the thyroid gland of the Balb-c mouse. In these mice TSH does not bind to its receptors, leading ultimately to the development of primary hypothyroidism, which is transmitted as an autosomal recessive trait. A maturational delay in the lung ultrastructure of the hyt/hyt mouse fetus has been observed. This investigation was undertaken to study the effect of maternal TRH treatment on lung ultrastructural maturation in the hyt/hyt mouse fetus. If the effect of TRH is mediated via stimulation of fetal pituitary-thyroid axis, TRH treatment should not enhance lung maturity in the hyt/hyt fetus and vice versa. Adult hyt/hyt mice made euthyroid by triiodothyronine supplementation were mated to carry hyt/hyt pups. Saline or TRH (0.4 or 0.6 mg/kg/dose) was administered to the mother (i.p.) on d 16 and 17 (b.i.d.) and on d 18 of pregnancy 1 h before killing (term, approximately 20 d). The fetal lung electron micrographs were subjected to ultrastructural morphometric analysis of the number of lamellar bodies and glycogen/nuclear ratio in type II cells, and the alveolar/parenchymal ratio by Chalkley point counting with an interactive computerized image analyzer (Optimas, Bioscan). Fetal lungs exposed to the lower dose of TRH (n = 7) showed no significant difference in their ultrastructural maturation when compared with saline-treated controls (n = 5). However, fetal lungs exposed to a higher dose of TRH (n = 6) showed increased numbers of lamellar bodies per type II cell, an increase in the alveolar/parenchymal ratio, larger air spaces, thinner alveolar septa, presence of tubular myelin, and increased numbers of air-blood barriers. We conclude that the effect of TRH in accelerating fetal mouse lung maturation is at least in part mediated via stimulation of extra thyroidal pathways. PMID- 9357948 TI - Abnormal expression of pulmonary bombesin-like peptide immunostaining cells in infants with congenital diaphragmatic hernia. AB - Infants with congenital diaphragmatic hernia (CDH) have a high neonatal mortality and morbidity owing to lung hypoplasia and persistent pulmonary hypertension. Pulmonary neuroendocrine cells produce bombesin-like peptide (BLP), a peptide with growth factor-like properties involved in lung development. We examined the expression of BLP immunostaining in pulmonary neuroendocrine cells (PNEC), and in clusters of these cells called neuroepithelial bodies (NEB), in the lungs of three groups of infants: patients with CDH, newborns with lung hypoplasia due to other causes, and control subjects without lung abnormalities. Morphometric analysis included: 1) percent immunostained airways; 2) percent immunostained epithelium (i.e. frequency of PNEC and NEB); and 3) NEB size. Controls and infants with lung hypoplasia did not differ with respect to BLP immunostaining. The ipsilateral and the contralateral lungs in CDH had a similar BLP immunostaining pattern of PNEC and NEB. The BLP immunostaining varied between CDH cases, possibly due to the differences in clinical presentation. The mean NEB size was significantly increased in infants with CDH compared with the other two groups (p = 0.02). Some CDH cases with large NEBs also showed a high percentage of immunostained epithelium. Lung-body weight ratio correlated positively with percent immunostained airways, and negatively with the NEB size. We conclude that in lungs of CDH patients BLP immunostaining in PNEC and NEB differs from that of infants with lung hypoplasia due to other causes and controls. The increased BLP immunostaining observed in some cases of CDH might reflect a compensatory mechanism related to impaired lung development and/or failure of neuropeptide secretion during neonatal adaptation. PMID- 9357949 TI - Pulmonary deposition of salbutamol aerosol delivered by metered dose inhaler, jet nebulizer, and ultrasonic nebulizer in mechanically ventilated rabbits. AB - The deposition efficiency of three methods of aerosol delivery of salbutamol into lungs of ventilated rabbits was compared: 1) metered dose inhaler (MDI) with holding chamber (HC), 2) jet nebulizer (JN), and 3) ultrasonic (US) nebulizer. The latter system was tested using two different sized medication reservoirs, a large (20 mL) cup (US20) and a small (10 mL) cup (US10). After delivery of technetium-99m-labeled salbutamol aerosol, deposition in the lungs, trachea, and ventilator circuit were estimated by a gamma counter. Total pulmonary deposition [mean(SEM)] as a percentage of the prescribed drug was: MDI + HC 0.22(0.05)%; JN 0.48(0.05)%; US20 0.90(0.13)%; US10 3.05(0.49)%. Only the deposition from the US10 was statistically significantly higher than the other modes (p < 0.05). Dynamic scintigraphy showed that, among the nebulizers, the US10 continued to deliver medication for longer than either the JN or the US20. We conclude that the US10 appears to be more efficient in delivering aerosol to the lung in this rabbit model and merits further evaluation for clinical efficiency. PMID- 9357950 TI - Cloning of a novel maize endosperm-specific protein with partial sequence homology to a pollen surface protein. AB - A gene encoding a novel maize endosperm protein has been cloned and sequenced. The gene encodes a 43 135 Da polypeptide which is 42% identical over two segments of an alfalfa pollen protein sequence. The gene is expressed in developing endosperm tissue, and not in other tissues such as shoot, pollen, or embryo. PMID- 9357952 TI - Control analysis of enzyme mechanisms in terms of the classical steady-state description. AB - The algebraic rate equations of steady-state enzyme kinetics generally allow the derivation of analytical expressions for the distribution of rate control among the elementary reactions defined by the kinetic mechanism. It is shown that these analytical expressions complete the macroscopic (probabilistic) description of enzyme-catalysed chemical reactions without involving any assumption at the microscopic (molecular) level. By also surveying some directly relevant results of the graph theoretical treatment of enzyme mechanisms, it is shown that control analysis is an integral part of steady-state enzyme kinetics. PMID- 9357951 TI - Sequence analysis of the cDNA encoding the precursor of equinatoxin V, a newly discovered hemolysin from the sea anemone Actinia equina. AB - A cDNA encoding the 214-amino-acid (aa) precursor of equinatoxin V (EqtV) has been isolated from an Actinia equina cDNA library. The sequence of the mature toxin is preceded, as that of EqtII, by a signal peptide of 19 aa and a hydrophilic propeptide of 16 aa ending with a pair of basic residues. This is similar to the precursors of calitoxins from another sea anemone Calliactis parasitica and to those of some antimicrobial peptides of the magainin and dermaseptin families from vertebrates. The deduced aa sequence of the potential cell attachment Arg-Gly-Asp motif-containing EqtV shows 82% identity to that of EqtII. PMID- 9357953 TI - Structure and function of the low Mr phosphotyrosine protein phosphatases. AB - Phosphotyrosine protein phosphatases (PTPases) catalyse the hydrolysis of phosphotyrosine residues in proteins and are hence implicated in the complex mechanism of the control of cell proliferation and differentiation. The low Mr PTPases are a group of soluble PTPases displaying a reduced molecular mass; in addition, a group of low molecular mass dual specificity (ds)PTPases which hydrolyse phosphotyrosine and phosphoserine/threonine residues in proteins are known. The enzymes belonging to the two groups are unrelated to each other and to other PTPase classes except for the presence of a CXXXXXRS/T sequence motif containing some of the catalytic residues (active site signature) and for the common catalytic mechanism, clearly indicating convergent evolution. The low Mr PTPases have a long evolutionary history since microbial (prokaryotic and eukaryotic) counterparts of both tyrosine-specific and dsPTPases have been described. Despite the relevant number of data reported on the structural and catalytic features of a number of low Mr PTPases, only limited information is presently available on the substrate specificity and the true biological roles of these enzymes, in prokaryotic, yeast and eukaryotic cells. PMID- 9357954 TI - A novel thermostable inhibitor of trypsin and subtilisin from the seeds of Brassica nigra: amino acid sequence, inhibitory and spectroscopic properties and thermostability. AB - A novel thermostable protein inhibitor of trypsin and subtilisin, called BN, was isolated from the seeds of Brassica nigra. The purified protein gave a single band on SDS-PAGE, corresponding to a molecular mass of 15 500 +/- 1000 Da. The inhibitor is composed of two disulfide-linked polypeptide chains, consisting of 39 and 90 residues, respectively. The amino acid sequence of the two chains was determined by Edman degradation of peptides, isolated from enzyme hydrolysates with TPCK-trypsin, EndoLysC proteinase and a Glu-specific proteinase of reduced and vinylpyridinated protein samples. A segment of the 'heavy' chain, between residues 65 and 81, showed homology with the reactive site loop region of the 6 kDa trypsin inhibitors from Nicotiana alata. The basic residue in position 39 (N. alata) or 70 (napins) is conserved as arginine or lysine in all inhibitors from N. alata and in all napins hitherto sequenced. Probably, the two families of trypsin inhibitors have structurally similar reactive sites. BN exhibits an extremely high thermostability: CD measurements showed that during heating to 97 degrees C it preserves a considerable part of the polypeptide backbone folding. Studies on the fluorescence properties of the inhibitor BN in the absence and presence of neutral or ionic quenchers demonstrated that the intrinsic emission of this protein is dominated by a tryptophyl residue, buried in the interior of the protein matrix. 20% of the light absorbed by Tyr 63 of the 'heavy' chain is transferred to Trp 26 of the 'light' chain. PMID- 9357956 TI - Purification and characterization of adenine phosphoribosyltransferase from Saccharomyces cerevisiae. AB - Adenine phosphoribosyltransferase (APRT) from Saccharomyces cerevisiae was purified approximately 1500-fold. The enzyme catalyzes the Mg-dependent condensation of adenine and 5-phosphoribosylpyrophosphate (PRPP) to yield AMP. The purification procedure included anion exchange chromatography, chromatofocusing and gel filtration. Elution of the enzyme from the chromatofocusing column indicated a pI value of 4.7. The molecular mass for the native enzyme was 50 kDa; however, upon electrophoresis under denaturing conditions two bands of apparent molecular mass of 29 and 20 kDa were observed. We have previously reported the presence of two separate coding sequences for APRT, APT1 and APT2 in S. cerevisiae. The appearance of two bands under denaturing conditions suggests that, unlike other APRTs, this enzyme could form heterodimers. This may be the basis for substrate specificity differences between this enzyme and other APRTs. Substrate kinetics and product inhibition patterns are consistent with a ping-pong mechanism. The Km for adenine and PRPP were 6 microM and 15 microM, respectively and the Vmax was 15 micromol/min. These kinetic constants are comparable to the constants of APRT from other organisms. PMID- 9357955 TI - Aspartate 155 of human transketolase is essential for thiamine diphosphate magnesium binding, and cofactor binding is required for dimer formation. AB - Active human transketolase is a homodimeric enzyme possessing two active sites, each with a non-covalently bound thiamine diphosphate and magnesium. Both subunits contribute residues at each site which are involved in cofactor binding and in catalysis. His-tagged transketolase, produced in E. coli, was similar to transketolase purified from human tissues with respect to Km apps for cofactor and substrates and with respect to cofactor-dependent hysteresis. Mutation of aspartate 155, corresponding to a conserved aspartate residue among thiamine diphosphate-binding proteins, resulted in an inactive protein which could not bind the cofactor-magnesium complex and which could not dimerize. The results are consistent with the suggestion that aspartate 155 is an important coordination site for magnesium. In support of this interpretation, binding of cofactor by wild type apo-transketolase required the presence of magnesium. Additionally, monomeric apo-his-transketolase required both magnesium and cofactor binding for dimer formation. PMID- 9357957 TI - Novel seminal phospholipase A2 is inhibited by the major proteins of bovine seminal plasma. AB - The activity of a novel phospholipase A2, isolated from bovine seminal plasma, was completely inhibited by low concentrations of the major proteins of bovine seminal plasma (BSP proteins). The inhibition was observed towards micellar as well as liposomal phosphatidylcholine. By solid phase binding assay it was determined that the inhibition involved a reduced accessibility to the lipidic substrate. Since the BSP proteins interact with choline phospholipids and since this interaction can be prevented by low concentrations of free choline, the effect of choline on the inhibition was studied. Choline could only partially relieve, even at high concentrations, the phospholipase A2 inhibition. The reduced accessibility appears to proceed via two distinct interactions: binding to the substrate on one hand and to the enzyme on the other hand. These results indicate that the BSP proteins may act as spermatozoa stabilizing agents by preventing premature lipolysis of the sperm surface. PMID- 9357959 TI - Characterization of guanosine kinase from Brevibacterium acetylicum ATCC 953. AB - Guanosine kinase (GKase) activity was identified in a cell-free extract of Brevibacterium acetylicum ATCC 953. We have purified the enzyme 4000-fold from the cell-free extract to apparent homogeneity. Molecular weight of 71,300 and 36,300 determined by gel filtration and sodium dodecyl sulfate gel electrophoresis, respectively, revealed that the enzyme is a dimer molecule. Maximum activity of the GKase was attained when the magnesium/ATP concentration ratio was close to 1. The GKase activity was dependent on the presence of a divalent cation. The Km values for guanosine, inosine, and 2'-deoxyguanosine as phosphate acceptors were determined to be 0.022, 0.87, and 2.83 mM, respectively. In addition to ATP and dATP, GTP and dGTP were shown to be effective phosphate donors for the GKase. The optimal pH seemed to be around pH 8.3, although relatively high activity was observed in the alkaline pH range of 7.5-9.8. The addition of 0.1 mM pyrimidine nucleotides, especially CMP and CTP, activated the GKase activity. On the other hand, the addition of 1 mM AMP, ADP, and GMP inhibited the GKase activity. It is thus likely that the GKase activity might be regulated in vivo by nucleotide concentrations and may control the nucleoside monophosphate level by efficiently recycling guanosine. PMID- 9357958 TI - Protective protein in the bovine lysosomal beta-galactosidase complex. AB - Cathepsin A [EC 3.4.16.1], so called protective protein, occurs as an enzyme complex with lysosomal beta-galactosidase [3.2.1.23] and is involved in the stable enzymic expression of lysosomal sialidase [3.2.1.18]. In this study we investigated the enzymatic properties of cathepsin A in the bovine beta galactosidase complex and how it is involved in the molecular multiplicities of the beta-galactosidase and sialidase complexes. Bovine protective protein homologous to the human protein had a molecular weight of 48 kDa on SDS-PAGE and cathepsin A activity optimum around pH 6.0. It hydrolyzed dipeptide substrates composed of hydrophobic amino acids much faster than any other type of substrate tested. This specificity was found to be conserved from human to a non-mammal, chicken. Immunoprecipitation using an anti beta-galactosidase antibody demonstrated that cathepsin A is a component of both the sialidase and beta galactosidase complexes. The over 700 kDa sialidase complex depolymerized by a brief incubation at pH 7.5 and the sialidase was inactivated irreversibly via formation of an enzyme active smaller species of sialidase. The 669 kDa beta galactosidase complex dissociated reversibly into a 120 kDa beta-galactosidase and a 170 kDa cathepsin A, but the 120 kDa beta-galactosidase, free from the cathepsin A, formed a 260 kDa aggregate under the same conditions. Inactivation of cathepsin A by heat treatment did not affect its complex forming activity. The 170 kDa protective protein dissociated into a 50 kDa one at pH 7.5, which no longer formed the complex. These findings indicate that the 170 kDa protective protein could be the minimum unit required for in vitro reconstitution of the complex, and that its complex forming activity is carried in a heat-stable domain. Both beta-galactosidase and cathepsin A activities were labile under the dissociated condition, indicating that it physiologically stabilizes not only beta-galactosidase but also itself by forming the complex. PMID- 9357960 TI - Multivariate analysis of the association of bromocresol purple anion with bovine serum albumin. AB - The interaction of bromocresol purple (BCP) anions with bovine serum albumin (BSA) was investigated by principal factor analysis method, and reaction model, the number of molecular species and spectra of each component present in the reaction mixture were determined. The number of molecular species concerning the absorption intensity was three, including free BCP anion. Most part of the spectral change could be explained by the monomer binding of bromocresol purple anion (D) to serum albumin (P), a simple one step equilibrium, P + D = PD. A second type of association of BCP anions with serum albumin was also present, though in a small amount. Of six models tested which consisted of three or four molecular species, the sequential two step reaction model, P = PD = PD2, was the best model to explain the spectral data, and an existence of BCP anions as a dimer on the serum albumin was demonstrated. The dissociation constants were estimated at K1 = 1.6 x 10(-6) M for the first step and K2 = 1.2 x 10(-5) M for the second step. PMID- 9357962 TI - Phosphorylation and calcium influx are not sufficient for the activation of cytosolic phospholipase A2 in U937 cells: requirement for a Gi alpha-type G protein. AB - Differentiation with dibutyryl cyclic AMP (dBcAMP) of the human, premonocytic U937 cell line toward a monocyte/granulocyte-like cell results in the cell acquiring an ability to release arachidonate upon stimulation. In contrast, the calcium ionophore ionomycin was able to stimulate phospholipase C, as measured by inositol 1,4,5-trisphosphate formation, to equal extents in both undifferentiated and dBcAMP-differentiated U937 cells. The role and regulation of cytosolic phospholipase A2 (cPLA2) in the production of arachidonate in these cells when either the chemotactic peptide fMLP or ionomycin are used as stimulus were investigated. The ionomycin- and fMLP-stimulated release of arachidonate were sensitive to the cPLA2 inhibitor arachidonyl trifluoromethylketone (IC50 values of 32 and 18 microM, respectively), but were not inhibited by E-6 (bromomethylene)-tetrahydro-3-(1-naphthalenyl)-2 H-pyran-2-one, a bromoenol lactone inhibitor of the calcium-independent phospholipase A2. These results, coupled with the inhibition of ionomycin-induced arachidonate production by electroporation of differentiated cells to introduce an anti-cPLA2, demonstrate that the cPLA2 is the enzyme responsible for arachidonate release in differentiated cells. SDS-PAGE and immunoblot analysis of differentiated cells showed the cells to contain both phosphorylated and unphosphorylated forms of cPLA2 (ratio of about 2: 3). Surprisingly, undifferentiated cells contain 30% more enzyme than differentiated cells and contain a higher percentage (approximately 75%) of the phosphorylated in the absence of stimulation. The inability of undifferentiated cells to produce arachidonate is not due to insufficient intracellular calcium concentrations since ionomycin induces large (820-940 nM) influxes of intracellular calcium in both differentiated and undifferentiated cells. This demonstrates that phosphorylation of cPLA2 andan influx of intracellular calcium are not sufficient to activate the enzyme to produce arachidonate. Instead, activation of a pertussis toxin-sensitive Gi alpha type G-protein is required as evidenced by the production of arachidonate in undifferentiated cells stimulated with mastoparan, an activator of Gi alpha subunits, in combination with ionomycin. This activation of a Gi alpha-type G protein is independent of modulations of adenylyl cyclase activity since cellular cAMP levels were not modulated upon treatment with mastoparan and ionomycin. PMID- 9357963 TI - Theoretical study of the electrostatic and steric effects on the spectroscopic characteristics of the metal-ligand unit of heme proteins. 3. Vibrational properties of Fe(III)CN-. AB - The vibronic theory of chemical activation and quantum chemical calculations are applied to calculate the stretching vibrational frequency of cyanide, coordinated by the complex of ferric porphyrin with imidazole. The results show that the frequency of the stretching vibration of the cyanide strongly depends on its coordination geometry and is hardly affected by the electrostatic perturbations of reasonable magnitude. The comparison of these results with the experimental data on the cyanide complexes of different heme proteins and their models allows to elucidate the cyanide coordination geometry. The combined infrared and resonance Raman scattering experimental investigation of the cyanide and carbonyl complexes with the same heme protein is proposed to distinguish between the steric and electrostatic contributions to the heme-protein interaction. PMID- 9357961 TI - Active site modification of aldose reductase by nitric oxide donors. AB - Nitric oxide (NO) donors sodium nitrosoprusside (SNP), S-nitroso-N acetylpenicillamine (SNAP), and 3-morpholinosydnonemine (SIN-1) caused a time- and concentration-dependent loss of catalytic activity of recombinant human placental aldose reductase. Modification of the enzyme was prevented by NADPH and NADP and reversed partially by dithiothreitol (DTT) and sodium borohydride. The protection by NADPH was lost in the presence of both substrates (NADPH and glyceraldehyde), indicating that the enzyme becomes sensitive to inhibition by SNP during catalysis. Site-directed mutant form of the enzyme, in which active site cys-298 was substituted with serine (C298S) was not inactivated by NO donors, whereas, ARC80S and ARC303 were as sensitive as the wild type enzyme, indicating that inactivation of aldose reductase is due to modification of the active site at cys298. These results suggest that NO may be an endogenous regulator of aldose reductase, and consequently the polyol pathway of glucose metabolism; which has been implicated in the pathogenesis of secondary diabetic complications. PMID- 9357964 TI - GAS6, the ligand of Axl and Rse receptors, is expressed in hematopoietic tissue but lacks mitogenic activity. AB - GAS6, a gene previously identified as growth arrest specific, has been demonstrated to be the ligand of Axl, a novel tyrosine kinase receptor widely expressed in both normal and neoplastic hematopoietic tissue. We have observed previously that GAS6 mRNA was present in whole bone marrow. This preliminary finding prompted us to investigate the presence of GAS6 in hematopoietic tissue and the possible role of this molecule in controlling the proliferation of hematopoietic precursors. We report here that the protein GAS6 is diffusely present in hematopoietic tissue, both in stromal and in hematopoietic cells, and that, among these cells, positivity is observed in megakaryocytes and myelomonocytic precursors. Furthermore, our data suggest that GAS6 is not a growth factor for hematopoietic progenitors or stromal fibroblasts. Despite the fact that both the Axl receptor and its ligand, GAS6, are expressed in hematopoietic tissue, the biological role of their interactions remains to be determined. PMID- 9357965 TI - Expression of the human major vault protein LRP in acute myeloid leukemia. AB - Overexpression of a 110-kD protein (lung resistance-related protein [LRP]) may predict a poor response to chemotherapy in patients with acute myeloid leukemia (AML) and ovarian carcinoma. The LRP gene has recently been mapped to chromosome 16, close to the multidrug resistance-associated protein (MRP) gene. Seventy seven samples from 67 patients with AML were examined for expression of LRP, MRP, and multidrug resistance (MDR1) mRNA using a semiquantitative reverse transcription polymerase chain reaction (RT-PCR) assay. Results were compared with 29 normal samples (11 normal peripheral blood and 18 normal bone marrow). Thirty-three patients with untreated AML were evaluable for response to chemotherapy. Levels of LRP, but not of MRP or MDR1 mRNA, were significantly higher in eight patients who failed to achieve complete remission (CR) compared with 25 patients who achieved CR (p = 0.033). A positive correlation was demonstrated between LRP and MRP (R = 0.368, p = 0.001) and between MRP and MDR1 mRNA levels (R = 0.301, p = 0.01) in the 77 clinical samples analyzed. In AML samples, a significant difference in MDR1 mRNA levels was found between presentation (47 samples) and relapse (30 samples) (p = 0.031). No significant difference was seen in LRP mRNA levels between these two groups or in eight patients studied sequentially at both presentation and relapse. Thirteen samples (10 at presentation, 3 at relapse) were analyzed for LRP protein expression by flow cytometry. Eight (5 at presentation, 3 at relapse) displayed greater than 10% positive cells (range 15-86%). These data suggest that LRP gene overexpression may constitute a novel mechanism of multidrug resistance. PMID- 9357967 TI - Effect of recombinant canine stem cell factor, a c-kit ligand, on hematopoietic recovery after DLA-identical littermate marrow transplants in dogs. AB - We studied the effect of recombinant canine stem cell factor (rcSCF) on hematopoietic recovery, incidence of graft failure, graft-vs.-host disease (GVHD), and survival after marrow transplantation from dog leukocyte antigen (DLA)-identical canine littermates. Ten animals received 100 microg rcSCF/kg/day b.i.d. by subcutaneous injection on days 1 through 10 after 920 cGy total body irradiation and transplantation of a mean of 3.7x10(8) marrow cells/kg body weight. None of the dogs received GVHD prophylaxis. All animals showed hematopoietic engraftment. The median number of days to achieve 1000 neutrophils/mm3 was 9; 100 monocytes/mm3 were reached after 15 days, 500 lymphocytes/mm3 after 21 days, and 20,000 platelets/mm3 after 16 days. One animal developed GVHD involving skin, gut, and liver and died of bacterial pneumonia 21 days after transplantation. The remaining nine dogs were observed for a median of 37 days (range 29-84 days) posttransplantation until they were killed. Facial edema was seen in three dogs during the first 2-3 days of rcSCF administration. These results show that within the limits of this study it appears to be safe to administer SCF after DLA-identical littermate marrow transplants in dogs. Comparison with previously published data in the same model showed that neutrophil and monocyte recovery was significantly faster in dogs receiving SCF treatment compared with dogs without growth factor treatment (recovery to achieve 1000 neutrophils/mm3: median 9 days vs. 13 days, p = 0.002; recovery to 100 monocytes/mm3: median 15 days vs. 105 days, p = 0.0002). Otherwise, no significant differences were seen. Results obtained with SCF treatment were similar to those previously obtained in the same model with recombinant human granulocyte colony-stimulating factor (rhG-CSF) treatment except that recovery of lymphocytes to 500/mm3 appeared to be more rapid in G-CSF-treated dogs (median 15 days vs. 21 days, p = 0.03). PMID- 9357966 TI - The relative population sizes of megakaryocytic cells in mouse bone marrow as determined by mpl ligand responsiveness. AB - The relative population sizes of mpl ligand-responsive megakaryocytic cells were investigated, and all megakaryocytes grown in culture were assessed. Three groups were analyzed: 1) immature cells with the ability to form single mature megakaryocytes; 2) cells with the ability to divide once before forming megakaryocytes (doublets); and 3) progenitor cells with the ability to form colonies, i.e., to undergo both cytokinesis and maturation. Immature cells forming single megakaryocytes proved most sensitive to the mpl ligand. Committed megakaryocyte progenitors required approximately 30 times more mpl ligand to achieve maximum growth than did the immature megakaryocyte population. Similar numbers of committed megakaryocyte progenitors responded to interleukin (IL)-3 and to mpl ligand. The amplification potential of these progenitor cells responding to each growth factor was assessed by measuring the number of megakaryocytes per colony. In response to mpl ligand progenitor, cells generated smaller colonies, with most cell divisions completed at a signifcantly earlier time point compared with progenitor cells responding to IL-3. The growth of more primitive megakaryocyte progenitors was best achieved in combination with other growth factors, notably IL-3; mpl ligand alone was ineffective in this regard. A novel finding was the significant number of megakaryocytes that grew in culture as closely coupled pairs (doublets). Data reported indicate that doublet formation may be a result of detection and stimulation of immature megakaryocytes rather than the diminished mpl ligand responsiveness of a proportion of megakaryocyte progenitors. By combining the number of mpl ligand-responsive cells forming single megakaryocytes with those forming megakaryocyte doublets, it is estimated that the size of the immature megakaryocyte pool greatly exceeds previous calculations. Thus we conclude that the immature megakaryocyte population is significantly larger than previously estimated and that these cells are the most sensitive to mpl ligand. Accordingly, these cells are potentially crucial in bone marrow responsiveness to mpl ligand that results from acute thrombocytopenia, being capable not only of endomitosis and maturation but perhaps of cell division as well. PMID- 9357968 TI - Human T lymphotropic virus-type I Tax induction of CD21/Epstein-Barr virus receptor expression on T cells and its significance in leukemogenesis of adult T cell leukemia. AB - CD21, which is expressed on B cells, is also expressed on human T lymphotropic virus-type I (HTLV-I)-infected T cell lines. CD21 also serves as a receptor of Epstein-Barr virus (EBV). We evaluated the mechanism of CD21 induction on HTLV-I infected T cells and its clinical significance in the leukemogenesis of adult T cell leukemia (ATL). CD21 induction was detected at very low levels in T cell lines (Jurkat and CEM cells), and in non- or low-Tax-producing HTLV-I-infected T cell lines (Oh13T, S1T, and Su9T01 cells). In contrast, marked induction of CD21 was detected in high-Tax-producing HTLV-I-infected T cell lines (K3T, F6T, and MT 2). A Jurkat T cell clone stably transfected with tax-expressing cDNA expressed a significant amount of CD21 on the cell surface. These results strongly suggest that HTLV-I Tax induces CD21 on T cells. On two-color analysis, CD21 expression was detected in CD4+ T cells of the primary ATL cells from a subset of patients, suggesting that EBV infection may be associated with the leukemogenesis of ATL, at least in part. However, no genome of EBV was detected in the genomic DNA of six HTLV-I-infected T cell lines or the primary ATL cells separated from all patients, indicating the irrelevance of EBV infection to ATL leukemogenesis. PMID- 9357970 TI - Pharmacological purging of minimal residual disease from peripheral blood stem cell collections of acute myeloblastic leukemia patients: preclinical studies. AB - In this paper we describe an experimental model for ex vivo purging of contaminating tumor cells from peripheral blood stem cell (PBSC) collections obtained from patients with acute myeloblastic leukemia (AML). We studied the combination of the alkylating agent nitrogen mustard (NM; concentrations ranging from 0.25 to 1.25 microg/mL) and etoposide (VP-16; constant dose of 20 microg/mL), and the conventional cyclophosphamide (Cy)-derivative mafosfamide (concentrations: 20-175 microg/mL). THE AIMS OF OUR STUDY WERE: 1) To compare the toxicity of the purging protocols on bone marrow (BM) and circulating trilineage precursors collected from normal donors after priming with granulocyte colony stimulating factor (G-CSF) or after complete remission (CR) consolidation chemotherapy and G-CSF (leukemic patients); 2) to demonstrate the survival of very primitive hematopoietic progenitors (LTC-IC) in the peripheral blood (PB) and the BM after pharmacological treatment; and 3) to evaluate the antineoplastic efficacy of purging protocols on PBSC collections using 3 well-established leukemic cell lines. Our results demonstrated that the toxicity on BM and PB progenitor cells could be correlated with the complete killing of committed granulocyte-macrophage colony-forming units (CFU-GMs) and erythroid precursors (BFU-Es), a condition reached at the concentration of 1.5 microg/mL of NM (in addition to 20 microg/mL of VP-16) and 175 microg/mL of mafosfamide. Notably, early and late megakaryocyte progenitor cells (CFU-MKs and BFU-MKs, respectively) showed higher sensitivity to NM/VP-16, but not to mafosfamide, than did CFU-GMs and BFU-Es. The dose of NM capable of inhibiting 95% of CFU-MKs and BFU-MKs (ID95) was 0.75 microg/mL. After incubation with the same dose of NM, the recovery of CFU-GMs and BFU-Es was 20 +/- 8% SD and 25 +/- 10% SD, respectively (p < 0.05). Long-term liquid cultures showed the recovery of primitive hematopoietic cells after incubation with the highest concentrations of NM/VP-16 and mafosfamide, with no significant differences between PB and BM samples. Under the same experimental conditions, we observed a more than 5-log reduction of contaminating leukemic cell lines (i.e., K-562, KG-1, and HL-60). In conclusion, we demonstrated that NM/VP-16 and mafosfamide purging agents are capable of killing leukemic cell lines that contaminate leukapheresis products from patients with AML, whereas an acceptable proportion of primitive LTC-IC is spared. Moreover, despite the different kinetic and functional profile of mobilized and steady-state BM progenitors, we did not observe any difference in toxicity of antineoplastic agents on hematopoietic cells at different levels of differentiation. These data suggest that pharmacological strategies developed for eliminating minimal residual disease (MRD) from BM autografts can be effectively and safely applied to circulating stem cell harvests. PMID- 9357969 TI - Stem cell factor suppresses apoptosis in neuroblastoma cell lines. AB - Stem cell factor (SCF) is a glycoprotein growth factor produced by marrow stromal cells that acts after binding to its specific surface receptor, which is the protein encoded by the protooncogene c-kit. SCF synergizes with specific lineage factors in promoting the proliferation of primitive hematopoietic progenitors, and has been administered to expand the pool of these progenitors in cancer patients treated with high-dose chemotherapy. SCF and its c-kit receptor are expressed by some tumor cells, including myeloid leukemia, breast carcinoma, small cell lung carcinoma, melanoma, gynecological tumors, and testicular germ cell tumors. Previous studies of SCF in neuroblastoma have produced conflicting conclusions. To explore the role of SCF in neuroblastoma, we studied five neuroblastoma lines (IMR-5, SK-N-SH, SK-N-BE, AF8, and SJ-N-KP) and the neuroepithelioma line CHP-100. All lines expressed mRNA for c-kit and c-kit protein at low intensity as measured by flow cytometry, and secreted SCF in medium culture as shown by ELISA. Exogenous SCF did not modify 3H thymidine uptake in the neuroblastoma and neuroepithelioma cell lines. After 6 days' culture in the presence of anti-c-kit, the number of viable neuroblastoma cells was significantly lower than the control, and terminal deoxynucleotidyl transferase assay showed a substantial increase of apoptotic cells: The percentage of positive cells was 1-3% in the control lines, whereas in the presence of anti c-kit it varied from 29% of SK-N-BE to 92% of CHP-100. After 9 days' culture in the presence of anti-c-kit, no viable cells were detectable. These data indicate that SCF is produced by some neuroblastoma cell lines via an autocrine loop to protect them from apoptosis. PMID- 9357971 TI - Diamond-Blackfan anemia: expansion of erythroid progenitors in vitro by IL-9, but exclusion of a significant pathogenetic role for the IL-9 gene and the hematopoietic gene cluster on chromosome 5q. AB - Diamond-Blackfan anemia (DBA) is a congenital pure red blood cell aplasia that often requires lifelong transfusional therapy. Autosomal dominant and recessive inheritance have both been reported, suggesting genetic heterogeneity, but most cases occur sporadically. The origin of impaired erythropoiesis is unknown. Several erythroid growth factors have been thought to have a role in the pathogenesis of DBA. However, there is neither molecular nor clinical evidence for the involvement of erythropoietin (EPO), its receptor, stem cell factor (SCF), or interleukin (IL)-3, even if the addition of SCF to IL-3 and EPO does significantly increase the growth of erythroid progenitors in in vitro cultures in most patients. In this work we evaluated the possible role of another early acting erythroid growth factor, IL-9. We found that the addition of IL-9 to SCF, IL-3, and EPO further increases burst-forming unit-erythroid growth in in vitro cultures of those DBA patients who responded to SCF. To investigate the role of the IL-9 gene, we evaluated its segregation in 22 families with members who have DBA by using a polymorphic microsatellite located within its intron 4. Lod score analysis ruled out any statistically significant involvement of the IL-9 gene in the pathogenesis of DBA. Moreover, linkage analysis with 11 highly polymorphic markers spanning 5q31.1-q33.2 excluded this region, which is included in the major cluster of genes active in hematopoiesis of the human genome. PMID- 9357972 TI - Altered production of GM-CSF and IL-8 in cytomegalovirus-infected, IL-1-primed umbilical cord endothelial cells. AB - The human cytomegaloviruses (HCMVs) appear to have the potential to disrupt production of hematopoietic cytokines. We examined the production of granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-8 by cultured and CMV-infected human umbilical vein endothelial cells (HUVECs) and compared this production with that of uninfected cells. Endothelial cells are, among other things, an integral component of human bone marrow stroma, and are responsible for production of factors that modulate the proliferation and differentiation of human hematopoietic progenitors. HCMV infection increased the production of GM-CSF in IL-1-primed HUVECs without altering GM-CSF levels in infected but unprimed HUVECs. However, this same virus was capable of causing increased production of the inhibitory cytokine IL-8. Both the viral pellet and the cleared viral supernatant appeared to contribute equally to the increased IL 8 and GM-CSF production, because each of these preparations alone was capable of exerting only half the effect seen with whole virus preparations. That both live virus and soluble protein factors within the viral stock contributed to the enhancement in GM-CSF and IL-8 production was further confirmed by inactivation with either ultraviolet or heat treatment of the viral stocks. Although the identity of the factor within the HCMV stock that contributes to this effect remains unknown, studies conducted in the presence of neutralizing antibodies or polymyxin B ruled out a role for tumor necrosis factor-alpha, IL-6, or endotoxin, all known inducers of GM-CSF. These studies indicate that HCMVs can exert both direct and indirect effects on the production of the hematopoietic factor GM-CSF and the inflammatory/inhibitory cytokine IL-8. PMID- 9357973 TI - Transgenic mice expressing the tyr22 variant of murine DHFR: protection of transgenic marrow transplant recipients from lethal doses of methotrexate. AB - Expression of the arg22, drug-resistant variant of dihydrofolate reductase (DHFR) in hematopoietic cells has been demonstrated to confer resistance to methotrexate (MTX) in mice, even though this variant suffers from low catalytic activity. The recently reported tyr22 variant has the advantage of higher catalytic activity combined with significant resistance to MTX. To evaluate the resistance conferred by tyr22-DHFR in vivo, we generated several transgenic mouse lines carrying a tyr22-DHFR minigene regulated by its natural promoter. The transgene copy number in 11 lines ranged from 6 to 68 copies and ribonuclease protection analysis demonstrated that 4 of these lines expressed significant transgenic DHFR mRNA at 20 to 68% of the endogenous DHFR mRNA level. Marrow from 4 of the 11 lines conferred significant increases in MTX-resistance in comparison with normal marrow when transplanted into lethally irradiated recipients. The ability of the tyr22-DHFR transgenic marrow to confer MTX-resistance to bone marrow transplant (BMT) recipients did not correlate with the level of mRNA expression or the number of transgene copies. However, two lines (lines 11 and 15) that were most effective in maintaining normal hematocrit levels in BMT recipients receiving 1 mg/kg/day MTX exhibited the greatest ability to form MTX-resistant hematopoietic progenitor colonies in vitro. Furthermore, MTX dose escalation studies demonstrated that line 11 marrow conferred resistance in BMT recipients receiving up to 6 mg/kg/day MTX. Southern blot analysis of the BMT recipients 7 months posttransplantation showed a preponderance of transgenic donor-derived cells in bone marrow and spleen, as well as a surprisingly high level in the small intestine. These results indicate that tyr22-DHFR is likely to be superior to arg22-DHFR in conferring MTX-resistance in BMT recipients, illustrating its usefulness for chemoprotection during MTX chemotherapy and also potentially for in vivo selection of transduced cells in gene therapy trials. PMID- 9357974 TI - Neplanocin A, a potent inhibitor of S-adenosylhomocysteine hydrolase, potentiates granulocytic differentiation of acute promyelocytic leukemia cells induced by all trans retinoic acid. AB - Several neplanocin A analogs were synthesized and their growth-inhibiting and differentiation-inducing activities on myelogenous leukemia cells were examined. An adenosine kinase-ineffective analog of neplanocin A was effective in inducing differentiation, suggesting that phosphorylation of the nucleoside is not essential for inducing the differentiation of leukemia cells. Neplanocin A induced functional and morphological differentiation of HL-60 cells, but did not effectively induce differentiation of NB4, a cell line derived from a leukemia patient with t(15;17). However, these cells have been known to undergo granulocytic differentiation upon treatment with all-trans retinoic acid (ATRA), and are used as a model for differentiation therapy in acute promyelocytic leukemia. Preexposure of NB4 cells to low concentrations of neplanocin A greatly enhanced the ATRA-induced differentiation of the cells, whereas representative antileukemic drugs such as cytosine arabinoside and daunomycin did not enhance this differentiation. A clinical strategy that combines intermittent treatment with neplanocin A analogs and a low dose of ATRA may increase the clinical response and decrease the adverse effects of ATRA. PMID- 9357975 TI - Expression of protein-tyrosine phosphatases in the major insulin target tissues. AB - Protein-tyrosine phosphatases (PTPs) are key regulators of the insulin receptor signal transduction pathway. We have performed a detailed analysis of PTP expression in the major human insulin target tissues or cells (liver, adipose tissue, skeletal muscle and endothelial cells). To obtain a representative picture, all tissues were analyzed by PCR using three different primer sets corresponding to conserved regions of known PTPs. A total of 24 different PTPs were identified. A multiprobe RNase protection assay was developed to obtain a semiquantitative measure of the expression levels of selected PTPs. Surprisingly, PTP-LAR, previously suggested to be a major regulator of the insulin receptor tyrosine kinase, was expressed in extremely low levels in skeletal muscle, whereas the related receptor-type PTP-sigma and PTP-alpha were expressed in relatively high levels in all four tissues. The low levels of LAR PTP mRNA in skeletal muscle were further confirmed by Northern blot analysis. PMID- 9357976 TI - Muscle glycogen synthase translocates from the cell nucleus to the cystosol in response to glucose. AB - We have studied the intracellular localization of muscular glycogen synthase by fusing the green fluorescent protein (GFP) of the jelly-fish Aequorea victoria to the N-terminus of human muscle glycogen synthase (HMGS), and expressing the chimeric protein in C2C12, COS-1 cells, and primary cultured rat hepatocytes. In contrast to what we have recently found for the hepatic glycogen synthase (Fernandez-Novell et al. (1997) Biochem. J. 321, 227-231), the GFP/HMGS fusion protein is localized to the nucleus of the cell in the absence of glucose, and in the presence of the sugar it is essentially found in the cytosol. Insulin is not required for the translocation of the enzyme. PMID- 9357977 TI - Crystal structure of cytochrome P-450cam complexed with the (1S)-camphor enantiomer. AB - The crystal structure of cytochrome P-450cam complexed with the enantiomer (1S) camphor has been solved to 1.8 angstroms resolution and compared with the structure of the (1R)-camphor P-450cam complex. The overall protein structure is the same for both enantiomer complexes. However, the orientation of the substrates in the heme pocket differs. In contrast to (1R)-camphor, the (1S) enantiomer binds in at least two orientations. The major binding mode of (1S) camphor resembles the one of the (1R)-enantiomer in that there is a hydrogen bond between Tyr-96 and the quinone group of camphor, and the 10-methyl group points towards the I-helix. The binding differs in that C-5 is not at a position suitable for hydroxylation. In the other orientation (1S)-camphor is not hydrogen bonded, but C-5 is located suitably for hydroxylation. PMID- 9357978 TI - Mapping of protein-protein interactions between c-myb and its coactivator CBP by a new phage display technique. AB - We have developed a phage display technique for the mapping of protein-protein interaction sites and characterized the interaction between the c-myb proto oncogene product and its co-activator CBP. Arbitrary DNA segments of the c-myb gene were cloned into a modified phagemid which allowed for expression in all possible reading frames. The mini-library encompassing all functional domains of the protein was propagated as phages and screened with different bait proteins. Alignment of the sequences revealed that the amino acids 317-342 of Myb interact with the CBP protein. Furthermore, an intramolecular interaction of the N terminal Myb DNA binding domain with the C-terminus (amino acids 541-567) could be detected. PMID- 9357979 TI - Distinct patterns of all-trans retinoic acid dependent expression of HOXB and HOXC homeogenes in human embryonal and small-cell lung carcinoma cell lines. AB - The expression patterns of the class I homeogenes HOXB and HOXC clusters in the presence of retinoic acid (RA) were studied in two human small-cell lung cancer (SCLC) cell lines and compared to that of NT2/D1 embryonal carcinoma cells. Contrasting with the sequential 3'-5' induction of the HOX genes observed after RA treatment of embryonic NT2/D1 cells, in the SCLC cells the responding genes (induced or down-regulated) were interspersed with insensitive genes (expressed or unexpressed), while no genomic alteration affected the corresponding clusters. These findings imply that HOX gene regulatory mechanisms are altered in non embryonic SCLC cells, perhaps reflecting their ability to respond to more diversified stimuli, in relation with their origin from adult tissues. PMID- 9357980 TI - Crystallization and preliminary X-ray analysis of arrestin from bovine rod outer segment. AB - We present the first X-ray study of a member of the arrestin family, the bovine retinal arrestin. Arrestin is essential for the fine regulation and termination of the light-induced enzyme cascade in vertebrate rod outer segments. It plays an important role in quenching phototransduction by its ability to preferentially bind to phosphorylated light-activated rhodopsin. The crystals diffract between 3 angstroms and 3.5 angstroms (space group P2(1)2(1)2, cell dimensions a = 169.17 angstroms, b = 185.53 angstroms, c = 90.93 angstroms, T = 100 K). The asymmetric unit contains four molecules with a solvent content of 68.5% by volume. PMID- 9357981 TI - Ceramide-induced translocation of protein kinase C zeta in primary cultures of astrocytes. AB - The present research was undertaken to study the possible involvement of the atypical protein kinase C (PKC) zeta in ceramide signal transduction in primary cultures of rat astrocytes. As shown by Western blot analysis, translocation of immunoreactive PKCzeta to the particulate fraction occurred upon exposure of astrocytes to cell-permeable ceramide analogs or to exogenous sphingomyelinase. The particulate fraction may correspond to a perinuclear area, as indicated by immunocytochemical techniques. Furthermore, treatment of cells with N octanoylsphingosine led to an increased phosphorylation of PKCzeta. Results thus show that stimulation of PKCzeta may be one of the intracellular events triggered by activation of the sphingomyelin pathway. PMID- 9357982 TI - Different expression of adenylyl cyclase isoforms after retinoic acid induction of P19 teratocarcinoma cells. AB - We have investigated the adenylyl cyclase (AC) activity and gene expression in retinoic acid (RA)-primed murine P19 teratocarcinoma cells, which recapitulate in vitro the first stages of neuroectodermal formation. Here we show that the P19 stem cells possess a basal Ca2+/CaM-stimulated AC activity, which increases about 10-fold after RA induction. The rise of AC activity is associated with a stage specific up-regulation of AC2, AC5 and AC8 mRNAs and a down-regulation of AC3 mRNA. P19 cells provide a powerful model to investigate the role and specific regulation of AC isoforms during neuronal differentiation. PMID- 9357983 TI - Glucokinase is located in secretory granules of pancreatic D-cells. AB - We immunohistochemically examined the distribution of glucokinase in rat pancreatic islets. Glucokinase immunoreactivity under light microscopy was detected in the cytoplasm of somatostatin cells as well as in that of insulin cells. No specific immunoreactivity was detected in glucagon and pancreatic polypeptide cells. In somatostatin cells, glucokinase immunoreactivity was located by electron microscopy exclusively within secretory granules. PMID- 9357984 TI - Bactericidal activity of human lysozymes carrying various lengths of polyproline chain at the C-terminus. AB - The amphiphilic polypeptide polyproline having different chain lengths was connected to the C-terminus of human lysozyme by the recombinant DNA technique. The hydrophobicity of human lysozyme increased with increasing length of the polyproline chain. Although the bactericidal activity of wild-type lysozyme is limited to gram-positive bacteria and the hydrolytic activity of the mutant lysozyme decreased with increasing chain length of polyproline, the mutant lysozymes showed bactericidal activity to gram-negative bacteria and the activity increased with increasing hydrophobicity of the mutant enzyme. Experiments with Escherichia coli phospholipid liposomes revealed that the mutant human lysozymes dissipated the valinomycin-induced transmembrane electrochemical potential, and the dissipation increased with increasing hydrophobicity. The increased hydrophobicity of the mutant enzyme may induce interaction of lysozyme with the outer membrane and subsequent penetration into the inner membrane of E. coli, resulting in an increase of bactericidal activity. PMID- 9357985 TI - Affinity and folding properties both influence the selection of antibodies with the selectively infective phage (SIP) methodology. AB - We investigated which molecules are selected from a model library by the selectively infective phage (SIP) methodology. As a model system, we used the fluorescein binding single-chain Fv fragment FITC-E2, and from a 3D-model, we identified 11 residues potentially involved in hapten binding and mutated them individually to alanines. The binding constant of each mutant was determined by fluorescence titration, and each mutant was tested individually as well as in competitive SIP experiments for infectivity. After three rounds of SIP, only molecules with KD values within a factor of 2 of the tightest binder remain, and among those, a mutant no longer carrying an unnecessary exposed tryptophan residue is preferentially selected. SIP is shown to select for the best overall properties of the displayed molecules, including folding behavior, stability and affinity. PMID- 9357986 TI - Mitochondrial cytochrome c oxidase subunit IV is phosphorylated by an endogenous kinase. AB - This study was undertaken to identify novel mitochondrial membrane proteins that are potential targets for phosphorylation. Mitochondrial membranes were incubated in the presence of [gamma-32P]ATP and the Triton X-114 extractable protein was subjected to ion-exchange and polyacrylamide gel chromatography to purify a major phosphorylated protein of approximately 17000 Da. The determined peptide sequence of the purified phosphoprotein corresponded to a segment of cytochrome c oxidase subunit IV, an inner membrane protein of 17160 Da. The identity of the phosphoprotein was confirmed by two-dimensional electrophoresis and Western blotting. The results identify mitochondrial cytochrome c oxidase subunit IV as a protein which is phosphorylated by an endogenous kinase. PMID- 9357987 TI - Nitric oxide directly activates calcium-activated potassium channels from rat brain reconstituted into planar lipid bilayer. AB - Using the planar lipid bilayer technique, we tested whether NO directly activates calcium-activated potassium (Maxi-K) channels isolated from rat brain. We used streptozotocin (STZ) as NO donor, and the NO release was controlled with light. In the presence of 100-800 microM STZ, the Maxi-K channel activity increased up to 3-fold within several tens of seconds after the light was on, and reversed to the control level several minutes after shutting off the light. Similar activation was observed with other NO donors such as S-nitroso-N acetylpenicillamine and sodium nitroprusside. The degree of activity increase was dependent upon the initial open probability (P[init]). When the P(init) was lower, the activity increase was greater. These results demonstrate that NO can directly affect the Maxi-K channel activity, and suggest that the Maxi-K channel might be one of the physiological targets of NO in brain. PMID- 9357988 TI - Expression and analysis of heparin-binding regions of the amyloid precursor protein of Alzheimer's disease. AB - Deletion mutagenesis studies have suggested that there are two domains within APP which bind heparan sulphate. These domains have been cloned and expressed in the yeast Pichia pastoris. Both recombinant proteins bound to heparin. One domain (APP316-447) was further characterised by binding studies with peptides encompassing this region. Peptides homologous to APP316-346 and APP416-447 were found to bind heparin. Circular dichroism studies show that APP416-447 shifted towards an alpha-helical conformation in the presence of heparin. This study suggests that heparin-binding domains may lie within regions high in alpha helical structure. PMID- 9357989 TI - Cross-linked trimers of bovine ribonuclease A: activity on double-stranded RNA and antitumor action. AB - Trimers of bovine pancreatic RNase A were obtained by cross-linking native RNase A with dimethyl suberimidate. They degrade double-stranded RNA more efficiently than dimers and monomers of RNase A, and display significant cytotoxic and/or cytostatic actions against C4-I cells (a human cell line derived from squamous carcinoma of the uterus cervix). On the same cell line cross-linked dimers of RNase A appear to be ineffective. PMID- 9357990 TI - The effect of hydroxylation of linoleoyl amides on their cannabinomimetic properties. AB - As yet, the physiological significance of hydroxylation of anandamide and linoleoyl amides is unknown. Therefore, we investigated whether hydroxylation of ODNHEtOH and ODNH2 influences their binding abilities to the CB-1 receptor and whether it alters their reactivity towards a fatty acid amide hydrolase (FAAH) from rat brain. Neither the fatty acid amides nor their hydroxylated derivatives were able to displace the potent cannabinoid [3H]CP 55.940 from the CB-1 receptor (Ki > 1 microM). Hydroxylation of ODNHEtOH resulted in a strong reduction of the maximum rate of hydrolysis by a FAAH, but the affinity of FAAH for the substrate remained of the same order of magnitude. Hydroxylation of ODNH2 led to a decrease in the affinity of FAAH for the substrate, but its maximum rate of conversion was unaffected. Furthermore, hydroxylation of ODNHEtOH enhanced its capacity to inhibit competitively the hydrolysis of anandamide. The resulting prolonged lifetime of anandamide and other fatty acid amide derivatives may have a considerable impact on cellular signal transduction. PMID- 9357991 TI - Role of the constriction loop in the gating of outer membrane porin PhoE of Escherichia coli. AB - Porins form voltage-gated channels in the bacterial outer membrane. These proteins are composed of three identical subunits, each forming a 16-stranded beta-barrel. In this study, the role in voltage gating of a loop that forms a constriction within the pore was studied. The channel characteristics of mutant PhoE porins, in which the tip of the constriction loop was connected to the barrel wall, were determined. Whereas the properties of several mutant channels were changed, all of these channels could still be closed at high potential, showing that a gross movement of the constriction loop within the channel is not implicated in voltage gating. PMID- 9357992 TI - Inhibition of DNA synthesis and tube morphogenesis of cultured vascular endothelial cells by chondromodulin-I. AB - Cartilage is an avascular tissue, and exhibits anti-angiogenic properties. Cartilage extracts have been shown to contain an inhibitor for DNA synthesis in vascular endothelial cells in vitro. Here we purified the inhibitory activity in the 10-50 kDa fraction of guanidine extracts from fetal bovine epiphyseal cartilage, and found that the inhibitor was identical with chondromodulin-I (ChM I). Purified ChM-I inhibited tube morphogenesis of cultured vascular endothelial cells, as well as DNA synthesis. These results indicate that cartilage-specific glycoprotein ChM-I may participate in the maintenance of avascularity and anti angiogenic properties of cartilage. PMID- 9357994 TI - An extremely heat-stable extracellular proteinase (aeropyrolysin) from the hyperthermophilic archaeon Aeropyrum pernix K1. AB - An extracellular metalloproteinase, which we had designated aeropyrolysin, from the aerobic marine hyperthermophilic archaeon Aeropyrum pernix K1 (JCM 9820), was purified by ammonium sulfate precipitation, anionic exchange chromatography, and gel filtration chromatography. The purified enzyme was composed of a single polypeptide chain with a molecular mass of 52 kDa as determined by SDS-PAGE. The proteinase had a broad pH optimum (pH 5-9) with a maximal activity at pH 6-8 for azocasein hydrolysis. The optimum temperature for enzyme activity was 100 degrees C in the absence of 1 mM CaCl2 and 110 degrees C in the presence of 1 mM CaCl2. The enzyme activity was completely inhibited by EDTA and EGTA, indicating that it was a metalloproteinase. The enzyme was highly resistant to the denaturing reagents urea, guanidine-HCl, dithiothreitol, 2-mercaptoethanol and SDS. The enzyme also showed a high activity with the metalloproteinase specific substrate MOCAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2. The enzyme was extremely thermostable showing half-lives of 2.5 h at 120 degrees C and 1.2 h at 125 degrees C in the presence of 1 mM CaCl2. These results indicate that this enzyme is one of the most thermostable extracellular proteinases reported to date. PMID- 9357993 TI - Arachidonic acid, a principal product of Rac-activated phospholipase A2, stimulates c-fos serum response element via Rho-dependent mechanism. AB - Previously, we have reported that phospholipase A2 (PLA2) is one of the major downstream targets by which Rac GTPase mediates the activation of c-fos serum response element (SRE) in response to agonists such as EGF [FEBS Lett. 407 (1997) 7-12]. Thus, the potential activity of arachidonic acid (AA), a principal product of Rac-activated PLA2, on c-fos SRE stimulation has been suggested. Here, we provide evidence about the biological activity of AA on c-fos SRE activation. Further, we observed that co-transfection with expression plasmid of either RhoN19, a dominant negative RhoA mutant, or botulinum C3 transferase which inhibits Rho via ADP ribosylation, selectively repressed AA- or Rac-induced SRE activation, suggesting that Rho activity is critical for the signaling cascade of 'Rac-PLA2-AA' to c-fos SRE. Thus, Rac signaling to the nucleus appears to be, at least partly, mediated by a Rho-linked pathway and this Rac-Rho signaling connection is mediated by AA. In accordance with the role of Rho as a potential mediator of AA signaling to the nucleus, AA induces a rapid translocation of RhoA. PMID- 9357995 TI - Concomitant and hormonally regulated expression of trp genes in bovine aortic endothelial cells. AB - Recent findings have suggested that the vertebrate trp family of channel proteins is the structural basis for Ca2+ influx through the capacitative calcium entry (CCE) pathway. We have discerned, in bovine aortic endothelial cells, the concomitant expression of four such vertebrate genes: trp-1 (two splice variants), trp-3, trp-4 and trp-5. Exogenous hormones rendered dynamic effects on the transcript levels of these genes. Most notably, beta-estradiol significantly down-regulated trp-4 while trans-retinoic acid dramatically up-regulated trp-5; yet these hormones rendered little change in CCE. These findings suggest that the extent of a given trp channel's participation in CCE is not reflected in alterations of its transcript level. PMID- 9357996 TI - Role of glutathione and nitric oxide in the energy metabolism of rat liver mitochondria. AB - Previous studies have suggested that nitric oxide (NO) inhibited mitochondrial respiration. NO and/or its intermediate(s) react with various molecules, such as hemeproteins and free SH groups. The inhibitory effect of NO on mitochondrial respiration was decreased by exogenously added glutathione (GSH). However, a decrease of intramitochondrial GSH by pretreating animals with L-buthionine sulfoximine had no appreciable effect on the inhibitory effect of isolated mitochondria. Furthermore, the effect of NO was not affected by depleting free SH residues in mitochondria by N-ethylmaleimide. These results suggest that cytosolic but not intramitochondrial GSH might be an important factor that determines the NO-dependent regulation of mitochondrial energy metabolism. PMID- 9357997 TI - Tissue inhibitor of metalloproteinases (TIMP)-1, -2 and -3 in human endometrium during the menstrual cycle. AB - The extensive remodelling of the human endometrium throughout the menstrual cycle is accompanied by changes in production of matrix metalloproteinases, the activity of which can be inhibited by specific tissue inhibitors or by tissue inhibitors of metalloproteinases (TIMP)s with a 1:1 stoichiometry. This study immunolocalized TIMP-1, TIMP-2 and TIMP-3 in dated normal human endometrium across the menstrual cycle and examined cultured endometrial cells for their production. All three TIMPs were present in the major cellular compartments, luminal epithelium, glands, stroma, endothelial cells and vascular smooth muscle cells with the most intense immunoreactivity in the luminal epithelium. TIMP-1 and -3 were lower in the mid-to-late proliferative phase with a nadir of TIMP-3 particularly in the late proliferative phase. Decidualized stromal cells stained strongly positive for TIMP-1, -2 and -3. Cells of haematopoietic origin never stained. Monensin treatment of tissue resulted in accumulation of TIMPs in all cellular compartments but particularly of TIMP-1 in epithelium. Cultured endometrial stromal cells released more TIMP-1 than TIMP-2 or TIMP-3 into culture medium and all were increased following decidualization in vitro. Epithelial cells in culture produced less TIMPs than stromal cells, and only a few epithelial cells in each culture were immunopositive for TIMP-1. The ubiquitous distribution of TIMPs implicates them in maintenance of endometrial integrity, with changes in the matrix metalloproteinases without concomitant changes in TIMPs determining endometrial matrix degradation. PMID- 9357998 TI - RU486 inhibits synthesis of an endogenous inhibitor of cell arachidonate release from choriodecidua tissue. AB - Gravidin is a potent phospholipase A2 inhibitor that may contribute to the maintenance of human pregnancy. The aims of this study were to determine firstly, the site of gravidin synthesis within placental membranes and secondly, whether the antiprogestin compound, RU486, regulates synthesis. Membrane explants were taken from placentae, dissected and cultured in the presence of RU486, progesterone or inhibitors of DNA or protein synthesis and gravidin production was measured by a specific enzyme-linked immunosorbent assay. Production of gravidin was consistently greatest from choriodecidua compared with amnion and decidua. The antibiotics, cycloheximide and actinomycin D inhibited production of gravidin. The progesterone receptor antagonist RU486 at 10(-8) and 10(-7) M reduced gravidin production to 55 and 39% of the control value in membranes cultured after and before the onset of labour respectively. Progesterone at 10( 6) M stimulated gravidin production by 47% after the onset of labour. The increase in gravidin production was correlated with progesterone concentration (r = 0.47, F = 6.38, P = 0.019) in labour tissue. We conclude that the data are consistent with the hypothesis that gravidin production may be partially regulated by progesterone. PMID- 9357999 TI - mRNA expression of insulin-like growth factor-I (IGF-I) is suppressed and those of IGF-II and IGF-binding protein-1 are constantly expressed in the endometrium during use of an intrauterine levonorgestrel system. AB - Insulin-like growth factors (IGF-I and IGF-II) are believed to mediate and modulate steroid hormone actions in the endometrium. In this study we determined the effects of an intrauterine system (IUS), releasing 20 microg levonorgestrel (LNG) daily, on endometrial expression of mRNAs encoding IGFs and insulin-like growth factor binding protein (IGFBP)-1. In Northern blotting, IGF-I mRNA was undetectable in all endometrial specimens from women with an LNG-IUS (n = 11) and in pregnancy decidua, whereas several transcripts of 0.6-7.6 kb were detected in proliferative and secretory phase endometria. In contrast, mRNAs encoding IGF-II and IGFBP-1 were strongly expressed in pregnancy and in all endometrial samples from women with an LNG-IUS, but were undetectable in proliferative or early to mid-secretory phase endometria. Using the more sensitive reverse transcriptase polymerase chain reaction (RT-PCR) method, IGF-I and IGF-II mRNAs were detectable in all cycling endometria, in early pregnancy decidua and in LNG-exposed endometrium. IGFBP-1 mRNA was constantly expressed in LNG-exposed endometrium, in early pregnancy decidua and in premenstrual endometrium, but was undetectable in all specimens from proliferative or early to mid-secretory endometrium. Our data demonstrate that progestin treatment can affect the gene expression of endometrial growth factors in vivo. The consistent expression of mRNAs encoding IGF-II and IGFBP-1, with suppression of IGF-I mRNA in endometria exposed to LNG, suggests that this mode of hormone treatment can inhibit IGF-I action in the endometrium. If IGF-I mediates and modulates oestrogen action, suppression of IGF I mRNA may be one of the molecular mechanisms which accounts for the antiproliferative effects of progestogens on oestrogen-primed endometrium and the atrophy of endometrial epithelium resulting from use of an LNG-IUS. PMID- 9358000 TI - Sperm nitric oxide and motility: the effects of nitric oxide synthase stimulation and inhibition. AB - Nitric oxide (NO) is synthesized from L-arginine by a family of enzymes known as the nitric oxide synthases (NOS). We have recently shown a NOS similar to constitutive brain NOS (bNOS) and endothelial NOS (ecNOS) to be present in spermatozoa. The aim of this study is to investigate NO production by human spermatozoa and the effects of stimulation and inhibition of NOS. This was carried out using the Iso-NO, an isolated NO meter and sensor, which provides rapid, accurate and direct measurements of NO. Semen samples with normozoospermic and asthenozoospermic profiles were prepared using a direct swim-up technique. Basal concentrations of NO and stimulated NO production were measured after exposure to the calcium ionophore (A23187; 0.01-10 microM) a potent activator of constitutive NOS. NO production in human spermatozoa was significantly increased by the addition of A23187 30 seconds after stimulation. Furthermore, this response was greatly diminished by pre-incubating the samples with competitive inhibitors of L-arginine, the substrate for NOS, before treatment with calcium ionophore. In the presence of N(G)-nitro-L-arginine methyl ester (L-NAME), N(G) nitro-L-arginine (L-NA) or N(G)-methyl-L-arginine (L-NMMA; all at 10 microM), NO production was inhibited with a rank order of potency L-NAME > L-NMMA > L-NA which is in accordance with the inhibition of an endothelial type of constitutive NOS. PMID- 9358001 TI - Autometallographic demonstration of zinc ions in rat sperm cells. AB - An in-vitro technique for autometallographic (AMG) demonstration of chelatable zinc in electroejaculated sperm cells and spermatozoa from the epididymis is presented and the localization of zinc ions in rat spermatozoa is described. Sperm cells from caput epididymis showed zinc staining in all parts of the tail and a sparse, dispersed staining in the acrosome. Spermatozoa from cauda epididymis showed heavy staining in the acrosome but no staining in the tail, or post-acrosomal part of the sperm head. This distinct acrosomal AMG staining was also found in ejaculated spermatozoa, but additionally a segmentation of the tail was seen based on differences in staining intensity. The membrane penetrating chelator diethyldithiocarbamate (DEDTC) was found to block the AMG staining whereas calcium-EDTA, known not to pass through cell membranes, did not influence the staining, proving that the detected zinc ions are intracellularly located. Two different approaches for demonstrating the presence of a chelatable zinc pool at electron microscope levels are presented, and the ultrastructural presence of AMG grains located in the acrosome and in the mitochondria of the midpiece is demonstrated. It is postulated that an exchange of zinc ions takes place between the epididymal epithelium and the sperm cells as they pass along the epididymal duct. PMID- 9358002 TI - Developmental regulation of telomerase activity in human fetal tissues during gestation. AB - Telomerase is a ribonucleoprotein that adds hexanucleotide repeats (telomeres) to the ends of linear chromosomes, compensating for the loss of telomeric DNA which occurs with DNA replication. In humans, telomerase has been previously detected in germ-line tissues, blastocysts, 16-20 week old fetal tissue, and most cancers, but not in mature sperm or ova, or in most normal somatic tissues. It has been hypothesized that telomerase is suppressed during somatic development and reactivated in malignancy. To test the hypothesis that telomerase is suppressed during somatic development, human fetal tissues of 8-21 weeks gestational age were assayed for telomerase activity. All tissues expressed telomerase at the earliest ages examined. Lung, liver, spleen, and testis maintained telomerase activity through the latest age assayed, namely 21 weeks. Brain and kidney telomerase activity was present up to the 16th week and was undetectable thereafter. Heart tissue did not display activity beyond the 12th week. Lysates of heart, brain, and kidney without telomerase activity did not inhibit the activity of known telomerase-positive cells, suggesting that suppression of telomerase activity during gestational development is due to a lack of active telomerase rather than to the presence of an inhibitor. These findings demonstrate tissue-specific and developmental regulation of telomerase in the human fetus, suggesting an important role for this ribonucleoprotein in human fetal tissue differentiation and development. PMID- 9358004 TI - Differential regulation of the plasminogen activator inhibitor-1 (PAI-1) gene expression by growth factors and progesterone in human endometrial stromal cells. AB - Plasminogen activator inhibitor-1 (PAI-1) has important regulatory functions in haemostasis, extracellular matrix turn-over and cell adhesion. We studied PAI-1 gene expression in primary cultures of endometrial stromal cells, and found that PAI-1 protein and mRNA were increased both by agents associated with differentiation, i.e. progesterone and transforming growth factor beta1 (TGFbeta1), and by those promoting proliferation, i.e. epidermal growth factor (EGF), TGFalpha and basic fibroblast growth factor (bFGF). In order to further elucidate the mechanism of regulation, we transfected stromal cells with an expression construct containing 804 bp of the PAI-1 promoter fused to a chloramphenicol acetyl transferase (CAT) reporter gene. After stimulation with the polypeptide growth factors TGFbeta1, EGF and bFGF we found increased CAT activity, indicating that these stimulators had initiated interaction with the transfected promoter fragment. On the other hand, stimulation with progesterone did not increase CAT activity, even though these cells were perfectly able to respond with increased secretion of PAI-1 protein. Run off experiments demonstrated that progesterone increased the stability of PAI-1 mRNA in endometrial stromal cells. We conclude that the polypeptide growth factors TGFbeta1, EGF and bFGF increase PAI-1 expression by increasing gene transcription. Progesterone, on the other hand, does not interact with the 804 bp promoter region, but increases the stability of PAI-1 mRNA. PMID- 9358003 TI - Glutathione S-transferase M1 gene polymorphism and susceptibility to endometriosis in a French population. AB - Endometriosis is a multifactorial disease with possible genetic predisposition and involvement of environmental factors in its pathogenesis. The genetic polymorphism of glutathione S-transferase M1 (GSTM1) gene, which codes for glutathione S-transferase 1, class mu foreign compound conjugating enzyme of phase II detoxification system, was studied by polymerase chain reaction from the blood spots in patients with different stages of endometriosis (n = 50) and in controls (n = 72) of French origin. A total of 86.0% of patients appeared to lack GSTM1 enzyme activity due to the presence of an extended deletion (GSTM1 0/0 genotype), compared with 45.8% in a control group (P < 0.0001), which was consistent with the frequency of GSTM1 deletion in French population. Moreover, the distribution of GSTM1-active genotypes was significantly different in patients and controls (P < 0.0001), as no patient with GSTM1A/B genotype, which is correlated with the highest activity of GSTM1 enzyme, has been found so far (18.1% in a control group). The unusually high frequency of homozygotes for the GSTM1 gene deletion among patients with endometriosis suggests a possible contribution of environmental toxins in the pathogenesis of this disease due to the absence or low activity of GSTM1 enzyme. PMID- 9358005 TI - Regulation of the human leukaemia inhibitory factor (LIF) promoter in HEC-1B endometrial adenocarcinoma cells. AB - Leukaemia inhibitory factor (LIF) is a pleiotropic cytokine which has been found to be expressed in the human endometrium and to play an important role in human reproduction. In the present study we investigated expression and regulation of the human LIF promoter in HEC-1B endometrial adenocarcinoma cells using a luciferase reporter plasmid bearing a 666 bp promoter fragment (h666LIF-Luc) in transient transfection assays. HEC-1B cells were first shown by reverse transcription-polymerase chain reaction (RT-PCR) to be able to produce endogenous LIF mRNA. The LIF promoter was efficiently transcribed in HEC-1B cells, showing much higher levels of basal activity than in the previously studied Jurkat T lymphoma cells and SKUT-1B uterine mesodermal tumour cells. The activity of the LIF promoter was stimulated in HEC-1B cells by a combination of phorbol ester (TPA) and ionomycin, which we had previously found to strongly induce its activity in Jurkat T-lymphoma cells. We next studied the effect of progestin (medroxyprogesterone acetate; MPA) on the LIF promoter activity in HEC-1B cells. The LIF promoter was not stimulated by MPA treatment in the presence of transfected progesterone receptor B (PR-B) expression vector in HEC-1B cells, while we had previously described its induction by MPA in SKUT-1B cells. This indicates that progestin-dependent regulation of the LIF promoter in uterine tumour cells is different in cells of epithelial and mesodermal origin. PMID- 9358006 TI - The cortex-medulla oocyte growth pattern is organized during fetal life: an in vitro study of the mouse ovary. AB - The cortex-medulla growth pattern of oocytes in the developing ovary is formed by a rim of small non-growing oocytes at the periphery and growing oocytes at the inner part of the cortex and in the medulla. In this study we aimed to: (i) develop an in-vitro model using fetal mouse ovaries to evaluate when the cortex medulla growth pattern is organized during ovarian differentiation; and (ii) study the interaction between small, non-growing and growing oocytes. Fetal mouse ovaries of days 11, 13 and 16 post coitus (E11, E13 and E16) were cultured for 14, 12 and 9 days respectively, corresponding to post-partum day 7 (P7). A cortex medulla growth pattern had developed by P7 in most ovaries from E16 and in half of those from E13. No ovaries from E11 developed a cortex-medulla pattern. The organization of this pattern within the ovary is concurrent with the initiation of meiosis. Ovaries with a cortex-medulla growth pattern had the same number of growing follicles (approximately 87), regardless of the number of small, non growing oocytes present. In contrast, in most ovaries without a cortex-medulla pattern, almost all oocytes began to grow. Hence, the geographically determined growth pattern of oocytes in the developing mouse ovary may be organized at the onset of meiosis and is sustained by a balanced interaction between small and growing follicles. PMID- 9358007 TI - Cloning and sequence analysis of rat fertilin alpha and beta--developmental expression, processing and immunolocalization. AB - Fertilin alpha and beta are members of the MDC (metalloproteinase-like, disintegrin-like, cysteine-rich) protein family and are expressed on the sperm surface where they have been proposed to play a role in mammalian fertilization. Inhibition of sperm-oocyte binding and sperm-oocyte fusion make fertilin an attractive target for the development of an immunocontraceptive vaccine. Full length cDNAs encoding alpha and beta fertilin subunits were isolated from a rat testis cDNA library and sequenced. Using reverse transcription-polymerase chain reaction (RT-PCR), the developmental expression of fertilin alpha and beta was determined in pre-pubertal and mature rat testes. Fertilin alpha mRNA was present at all stages of development, suggesting that it is not exclusively expressed in post-meiotic germ cells. In contrast, fertilin beta mRNA was first identified in day 19 testes, coincident with the presence of pachytene spermatocytes. Polyclonal antisera raised against a 28-residue peptide (corresponding to part of the disintegrin domain) and two recombinant fusion proteins identified a 90 kDa protein in testicular sperm extracts and a 60 kDa protein in caput and cauda epididymidal sperm extracts, the predicted sizes for rat fertilin beta precursor and mature protein respectively. Indirect immunofluorescence using the anti peptide antisera stained the acrosomal cap of permeabilized testicular, caput and caudal spermatozoa and elongating spermatids in testicular sections. PMID- 9358008 TI - Oligoasthenospermia associated with multiple mitochondrial DNA rearrangements. AB - A patient who wished to be treated for infertility by intracytoplasmic sperm injection (ICSI) was referred to our group for assessment. Upon clinical examination, a ptosis (partial closure of the eyelid) was noted, and histology revealed ragged red fibres in the skeletal muscle. Southern blot analysis of spermatozoa and skeletal muscle revealed the presence of multiple mitochondrial DNA deletions. This kind of rearrangement may be of nuclear origin since three nuclear loci have been ascribed to multiple mitochondrial DNA deletions in humans. Since mitochondrial DNA is maternally transmitted, the use of ICSI was feasible. However, an alteration of nuclear gene product affecting the integrity of mitochondrial DNA, and thus sperm mobility, might be transmitted to the offspring with the risk of developing a mitochondrial DNA disease. PMID- 9358009 TI - Estimation of aneuploidy for chromosomes 3, 7, 16, X and Y in spermatozoa from 10 normospermic men using fluorescence in-situ hybridization. AB - Fluorescence in-situ hybridization (FISH) is a fast and efficient method of estimating aneuploidy in human spermatozoa. In this study, we have estimated baseline disomy frequencies in spermatozoa from a group of 10 normospermic men, using stringent scoring criteria. A triple-probe FISH procedure was used for chromosomes 3, X and Y, while a double-probe FISH method was used for chromosomes 7 and 16. A total of 101273 spermatozoa were scored for chromosomes 3, X and Y, resulting in 97.83% haploidy (3X or 3Y), 0.39% disomy (33X, 33Y, 3XX, 3YY or 3XY) and 0.35% diploidy (33XX, 33YY or 33XY). A total of 100760 spermatozoa were scored for chromosomes 7 and 16, giving 98.9% haploidy (716), 0.11% disomy (7716 or 71616) and 0.27% diploidy (771616). Disomy frequencies for individual chromosomes differed (chromosome 3, 0.20%; chromosome 7, 0.05%, chromosome 16, 0.06%; X + Y, 0.19%). The frequency of disomy 3 was significantly higher than disomy 7 (P = 0.019) and disomy 16 (P = 0.022), while the frequency of sex chromosome disomy was significantly higher than disomy 7 (P = 0.0058) and disomy 16 (P = 0.0067), but not disomy 3 (P = 0.73). The disomy and diploidy (0.27 0.35%) estimates obtained for this normospermic population were generally low and were similar to other recent reports. PMID- 9358010 TI - Hoechst staining and exposure to UV laser during flow cytometric sorting does not affect the frequency of detected endogenous DNA nicks in abnormal and normal human spermatozoa. AB - Controlling the sex of offspring by the separation of X and Y chromosome-bearing spermatozoa using flow cytometry has been reported as a clinical technique aiding prevention of X-linked diseases. Although this technique has resulted in several hundred normal births in animals and at least one human birth, there is still concern over its genetic safety due to the involvement of two potentially mutagenic agents: UV light and the fluorochrome dye, Hoechst 33342 (H33342). Human spermatozoa, particularly those considered abnormal, may be more likely to suffer DNA damage following exposure to mutagenic agents, compared with other mammalian species. The stability of normal fresh and decondensed human spermatozoa were examined after exposure to a range of levels of UV and H33342 staining, using an assay that detects endogenous nicks in the DNA of spermatozoa. The stability of abnormal and normal, fresh and frozen-thawed human spermatozoa was examined following UV laser, H33342 staining and flow cytometry treatments utilizing the same assay. There was an increase in the presence of endogenous nicks when spermatozoa were decondensed compared with fresh spermatozoa. There was no increase in the incidence of nicks in any group of spermatozoa after UV and fluorochrome exposure compared with controls without exposure. PMID- 9358011 TI - The spectrum of beta-thalassaemia mutations in the Lebanon. AB - We screened 110 DNA samples from carriers of beta-thalassaemia, using the ARMS PCR technique with primers for common Mediterranean mutations. Unidentified samples were subjected to a heteroduplex analysis with Universal Heteroduplex Generators covering the beta-globin gene, followed by DNA sequencing. In total, 16 different mutations were detected, the most frequent being IVSI-110 (40%), followed by other common Mediterranean mutations (IVSI-1, IVSII-1, IVSI-6). Other mutations detected were of Lebanese, Turkish, Iranian, Kurdish, Bulgarian and Asian Indian origin. The most heterogeneous religious group seems to be the Sunni Muslims, with 13 mutations, while only 2 mutations were detected among the Christian Maronites. Results from this study are compared with those from other Mediterranean and neighbouring countries. PMID- 9358012 TI - Angiotensin-converting enzyme deletion polymorphism is associated with hypertension in a Sikh population. AB - The deletion polymorphism, situated in intron 16, of angiotensin-converting enzyme (ACE) gene (17q23) has been observed to be associated with an increased risk for myocardial infarction and left ventricular hypertrophy in Caucasian populations. The homozygous genotype for the deletion allele (DD) has additionally been observed at greater frequencies in hypertensive individuals of African-American and Japanese origin. In a population-based study of a Sikh population, we compared the occurrence of the insertion/deletion polymorphism at the ACE gene in subjects with hypertension to those with normal blood pressure. The ACE deletion allele was observed with a greater frequency in hypertensive subjects than in the normotensive subjects (p < 0.0001). These findings raise the possibility that in some ethnic subgroups, variation in or near the ACE gene may contribute to the development, and severity, of hypertension. PMID- 9358013 TI - Molecular and cytogenetic investigations of the fragile X region including the Frax A and Frax E CGG trinucleotide repeat sequences in families multiplex for autism and related phenotypes. AB - We undertook molecular and cytogenetic analyses in 25 families multiplex for autism and related disorders. Three of the multiplex families exhibited fragile X, and the affected offspring all exhibited CGG triplet repeat insertion mutations in the FMR-1 gene. One of these families contained an affected pair of monozygotic female twins. Both had similar-sized CGG triplet repeat expansions, but different phenotypic manifestations. One suffered from autism and the other from mild mental retardation and marked social anxiety. PCR and Southern hybridization analysis of the CGG repeat sequences characterizing fragile X A (Frax A) and E and the methylation status of FMR-1 showed no evidence of abnormal CGG repeat expansion or FMR-1 hypermethylation in the remaining 22 multiplex families. Moreover, there was no correlation between the Frax A or E (CGG)n repeat length with affected status, nor any association with the low-level (< 3 %) expression of cytogenetic fragility at Xq27 previously reported in these families. Our findings indicate that most instances of recurrence in families multiplex for autism and related disorders are not accounted for by Frax A and E. They also indicate that the phenotypic manifestations of Frax A may be influenced by stochastic, environmental and other biological factors. PMID- 9358014 TI - Ethnic differences in the HFE codon 282 (Cys/Tyr) polymorphism. AB - Recent studies have shown that hereditary hemochromatosis (HH) is likely to be caused by homozygosity for a Cys282Tyr mutation in the HFE gene located 4.5 Mb telomeric to HLA-A. Population studies of this polymorphism are facilitated by the fact that the Cys282Tyr mutation creates a Rsal restriction site. We have studied the codon 282 (Cys/Tyr) polymorphism in different ethnic groups. In agreement with previous observations the Tyr allele appeared to be rare or absent in Asiatic (Indian, Chinese) populations. The highest allele frequency (7.5%) was found in Swedes. Saamis (2%) and Mordvinians (1.8%) had significantly lower frequencies of the Tyr allele. Comparisons with allele frequencies based on prevalence estimates of HH showed some disagreements with the RFLP data, particularly in Finns. The newly described HFE marker provides a new approach to the screening of HH as well as studies of the relationship between the HFE Tyr allele and different disorders including cancer. PMID- 9358015 TI - Linkage analysis of manic depression (bipolar affective disorder) in Icelandic and British kindreds using markers on the short arm of chromosome 18. AB - Attempts were made to follow up results of a previous linkage study which suggested that a locus-modifying susceptibility to bipolar and related unipolar affective disorder might be present in the pericentromeric region of the short arm of chromosome 18. Twenty-three multiply affected pedigrees collected from Iceland and the UK were genotyped using three highly polymorphic microsatellite markers at D18S37, D18S40 and D18S44 which span the region implicated. Lod score analyses under the assumption of heterogeneity and non-parametric linkage analyses were performed. The total lod scores obtained were strongly negative, and analysis allowing for heterogeneity did not suggest that any subgroup of the families was linked. Model-free linkage analysis using extended relative pair analysis and MFLINK also failed to detect any evidence for linkage. Our study provides no support for the presence of a locus-modifying genetic susceptibility to bipolar affective disorder in the pericentromeric region of chromosome 18q11. Further analyses in independent samples should help to reveal whether our negative results are due to locus heterogeneity or whether the original results were false-positive. PMID- 9358016 TI - Investigation of complement C4B deficiency in schizophrenia. AB - Several lines of evidence suggest that autoimmune mechanisms might contribute to the development of schizophrenia. Important factors involved in immune responses in man include the human leukocyte antigens and components of the complement system. In the present study we attempted to confirm a positive association between a homozygous deficiency in complement factor C4B and schizophrenia as previously reported. We also determined parental genotypes in a subset of our schizophrenic patients to test the hypothesis of a genetic mechanism depending on the mother's genotype. C4B deficiency was found in similar frequency among patients (n = 176) and controls (n = 145). There was also no increased frequency of C4B deficiency in the mothers of schizophrenic patients. Our study does not support a widespread or consistent association between a deficiency in complement component C4B and schizophrenia. PMID- 9358017 TI - Haptoglobin polymorphism in Korean patients with cardiovascular diseases. AB - We investigated the relation between hepatoglobin (Hp) polymorphism and plasma lipid levels in 913 Korean subjects. The distribution of Hp phenotypes did not show any significant differences between the healthy controls and the patients with cardiovascular disease. In the control group, however, the subgroup of > or = 50-year-olds had a significantly higher Hp*1 allele frequency than the subgroup <50 years (p < 0.005). This was not seen in the patient group. Hp phenotypes were associated with levels of high-density lipoprotein cholesterol in the hypertensive group. The results indicate that Hp polymorphism, at least in the Korean population, does not predispose to the occurrence of cardiovascular diseases. PMID- 9358018 TI - Paternity analysis in cases of father-daughter incest using multiallelic loci. AB - The inclusion of DNA polymorphism into paternity analysis represents a major advance since it allows one to achieve a high probability of exclusion. This is of particular importance in paternity cases in which the mother and the alleged father are close relatives. The purpose of this article is to demonstrate an approach to calculate the probability of exclusion and to compute the paternity index in case of father-daughter incest using data from multiallelic loci that can be obtained with a single locus probe. The global probability of exclusion achieved at a multiallelic locus, in cases of father-daughter incest, is lower than the one obtained when both parents are not relatives. The probability of exclusion of each mother-offspring pair is different from that obtained when the parents are not close relatives. Among mother-offspring pairs having the same genotype, the probability of exclusion is zero, and in such a condition, the locus is not informative. To obtain a high exclusion capability, it is necessary to analyze several independent loci in which both the mother and the child have different genotypes. Expressions to obtain the paternity index for all the possible combinations of mother-offspring-alleged father in cases of father daughter incest are derived and discussed. PMID- 9358019 TI - A CA repeat in the first intron of the CFTR gene. AB - Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, that encompasses 250 kb of genomic DNA, cause cystic fibrosis. More than 5-10% of CF patients in most populations studied carry undefined mutations and hence intragenic CA repeats are important tools in genetic counselling. To date, polymorphic intragenic repeats have been found in introns 6a, 8 and 17b. We have identified a novel CA repeat within intron 1 of the CFTR gene that lies about 70 kb 5' to intron 6a and so will be a useful additional diagnostic marker. PMID- 9358020 TI - Association between allele 1 of T102C polymorphism, 5-hydroxytryptamine 2a receptor gene and schizophrenia in Chinese males in Singapore. AB - The T102C polymorphism at the 5-hydroxytryptamine 2a receptor (5HTR2a) gene was studied in 101 Chinese male schizophrenics and 103 controls. Genotyping revealed a predominance of allele 1 among schizophrenics. This is in contrast to previous reports in Caucasians and Japanese which showed an association of allele 2 of this polymorphism with schizophrenia. PMID- 9358022 TI - Naked DNA gene delivery to the liver. PMID- 9358021 TI - Bystander effects in enzyme/prodrug gene therapy. PMID- 9358024 TI - Retrovirus-mediated gene transfer of the multidrug resistance-associated protein (MRP) cDNA protects cells from chemotherapeutic agents. AB - Transduction of hematopoietic progenitors with a multidrug resistance gene like mdr-1 or mrp aims to protect bone marrow from toxicity of chemotherapeutic agents. The interest in the use of mrp as an alternative to mdr-1 gene transfer for bone marrow protection lies in its different modulation. Indeed, classical P gp reversal agents, tested in the clinic to decrease mdr-1 tumor resistance, have little or no effect on MRP function. This would allow, in the same patient, the use of reversal agents to decrease P-gp tumor resistance without reversing bone marrow protection of the transduced hematopoietic cells provided by multidrug resistance-associated protein (MRP). As a first step, we have constructed and tested two different mrp-containing vectors with either the Harvey retroviral long terminal repeat (LTR) or PGK as promoters and generated ecotropic producer cells. We have shown by Southern blot analysis that retroviral supernatant from these producer cells can efficiently transmit the mrp gene to target cells. Mrp expression could be detected by fluorescence-activated cell sorting (FACS) analysis in the producer cells. The transduced cells have increased resistance to doxorubicin, vincristine, and etoposide. Furthermore, chemoprotection of the transduced cells was increased after selection with chemotherapeutic agents in the presence of glutathione, a co-factor for MRP function. These data indicate that mrp retroviral vectors may be useful for chemoprotection and selection. PMID- 9358023 TI - VEGF gene transfer reduces intimal thickening via increased production of nitric oxide in carotid arteries. AB - Thickening of the arterial intima and smooth muscle cell (SMC) proliferation remain major problems after vascular surgery and other types of vascular manipulations. We studied the effect of endothelial cell (EC)-specific vascular endothelial growth factor (VEGF) gene transfer on the thickening of the intima using a silicone collar inserted around carotid arteries that acted both as the agent that caused intimal SMC growth and as a reservoir for the transfected gene. The model preserved EC integrity and permitted direct extravascular gene transfer without any intravascular manipulation. Compared to beta-galactosidase (lacZ) transfected control arteries, plasmid/liposome-mediated VEGF gene transfer significantly reduced intimal thickening 1 week after the gene transfer. Administration to the experimental animals of the nitric oxide (NO) synthase inhibitor L-NAME abolished the difference in intimal thickening between VEGF and lacZ-transfected arteries. Furthermore, VEGF caused NO release from cultured human umbilical vein EC. It is concluded that extravascular VEGF gene transfer attenuates intimal growth and could be useful for the prevention of intimal thickening during vascular surgery. Our results further suggest that VEGF may reduce SMC proliferation via a mechanism that involves VEGF-induced NO production from the endothelium. PMID- 9358025 TI - Gene therapy in hypertension: adenovirus-mediated kallikrein gene delivery in hypertensive rats. AB - Tissue kallikrein has been shown to play a role in blood pressure regulation, and abnormalities in the kallikreinkinin system are considered to be a factor in the pathogenesis of hypertension. To elucidate the potential therapeutic effects of kallikrein gene delivery in hypertension, an adenoviral vector containing the human tissue kallikrein gene under the control of a cytomegalovirus promoter, Ad.CMV-cHK, was intravenously injected into spontaneously hypertensive rats (SHR). A single injection of Ad.CMV-cHK into SHR caused a sustained delay in the increase in blood pressure from day 2 to day 41 post injection, as compared to control rats receiving Ad.CMV-LacZ adenovirus. Adenovirus-mediated kallikrein gene delivery had no effect on the blood pressure of normotensive Wistar-Kyoto rats. Human tissue kallikrein mRNA was detected in the liver, kidney, spleen, adrenal gland, and aorta. Immunoreactive human tissue kallikrein can be detected in sera and urine of rats receiving kallikrein gene delivery. Human tissue kallikrein in rat serum was at the highest level 5 days post injection, and the level declined gradually. Urinary kinin and cGMP levels were significantly increased in rats receiving kallikrein gene delivery compared to Ad.CMV-LacZ control rats. These results show that adenovirus-mediated delivery of human tissue kallikrein results in high-efficiency expression and blood pressure reduction in SHR. Application of adenovirus-mediated systemic expression of the tissue kallikrein gene may provide a unique way of delivering the gene product into the vasculature and could have important therapeutic implications in treating hypertension. PMID- 9358026 TI - Expression of naked plasmid DNA injected into the afferent and efferent vessels of rodent and dog livers. AB - A variety of reporter genes within plasmid constructs were injected into the afferent and efferent vessels of the liver in mice, rats, and dogs. Efficient plasmid expression was obtained following delivery via the portal vein, the hepatic vein, and the bile duct. The use of hyperosmotic injection solutions and occlusion of the blood outflow from the liver substantially increased the expression levels. Combining these surgical approaches with improved plasmid vectors enabled uncommonly high levels of foreign gene expression in which over 15 microg of luciferase protein/liver was produced in mice and over 50 microg in rats. Equally high levels of beta-galactosidase (beta-Gal) expression were obtained, in that over 5% of the hepatocytes had intense blue staining. Expression of luciferase or beta-Gal was evenly distributed in hepatocytes throughout the entire liver when either of the three routes were injected. Peri acinar hepatocytes were preferentially transfected when the portal vein was injected in rats. These levels of foreign gene expression are among the highest levels obtained with nonviral vectors. Repetitive plasmid administration through the bile duct led to successive events of foreign gene expression. The integration of these findings into laboratory and clinical protocols is discussed. PMID- 9358027 TI - A bicistronic retroviral vector for protecting hematopoietic cells against antifolates and P-glycoprotein effluxed drugs. AB - Chemoresistance gene transfer is an experimental method to protect hematopoietic cells from the toxicity of anticancer drugs. Because multiple drugs are usually given together in cancer therapy, this strategy will ultimately require vectors expressing multiple chemoresistance genes. For this reason, we designed a bicistronic retroviral vector (HaMID) containing a modified human multidrug resistance-1 cDNA and a mutant human dihydrofolate reductase cDNA bearing a leucine to tyrosine substitution at codon 22 (L22Y). To determine if this vector would confer dual drug resistance to hematopoietic cells, recombinant retrovirus was used to transduce the human CEM T lymphoblastic cell line as well as primary murine myeloid progenitors. Growth suppression assays, using polyclonal transduced CEM cells, demonstrated increased resistance to taxol (13-fold), trimetrexate (8.9-fold), vinblastine (5.6-fold), methotrexate (2.5-fold), and etoposide (1.5-fold) when used as single agents. HaMID-transduced cells also grew at a logarithmic rate in the simultaneous presence of 25 nM taxol and 100 nM trimetrexate while control cells were entirely growth inhibited by this drug combination. Similarly, HaMID-transduced murine myeloid progenitors acquired increased resistance to taxol (2.9-fold) and trimetrexate (140-fold), and were able to form colonies in the simultaneous presence of both drugs. Our results suggest that retroviral transfer of HaMID into primary hematopoietic cells should reduce the myelosuppression associated with the combined use of antifolates and P glycoprotein-effluxed drugs. PMID- 9358028 TI - Expression of biologically active human insulin-like growth factor-I following intramuscular injection of a formulated plasmid in rats. AB - Recent evidence has shown that insulin-like growth factor-I (IGF-I) plays an important role in the development, maintenance, and regeneration of peripheral nerves and skeletal muscle. IGF-I offers the potential to treat neuromuscular diseases in humans. We have developed a nonviral gene therapy method to express and produce localized and sustained therapeutic levels of IGF-I within target muscles by intramuscular injection of formulated plasmids. The purpose of the present study was to demonstrate that intramuscular injection of a plasmid encoding human IGF-I (hIGF-I) and engineered to restrict expression to skeletal muscle produces sustained local concentrations of biologically active hIGF-I. Normal rats received a single intramuscular injection of plasmids formulated as a complex with polyvinylpyrrolidone (PVP). Results show that hIGF-I mRNA and hIGF-I protein were detectable in the injected muscles for the duration of the study (28 days), whereas the hIGF-I protein was not detected in blood. Biological activity of hIGF-I was determined by immunodetection of a nerve-specific growth-associated protein, GAP-43, an indicator of motor neuron sprouting. Placement of human growth hormone (hGH) 3' untranslated region enhanced GAP-43 staining, probably due to improved secretion of hIGF-I. Enhanced immunoreactivity of GAP-43 was observed in muscles injected with the formulated hIGF-I plasmid when compared to controls. These results demonstrate that intramuscular injection of hIGF-I plasmid formulated as a complex with PVP produces a localized and sustained level of biologically active hIGF-I. PMID- 9358030 TI - A "distant" bystander effect of suicide gene therapy: regression of nontransduced tumors together with a distant transduced tumor. AB - Antitumor gene therapy using herpes simplex type 1 thymidine kinase (TKh) and ganciclovir (GCV) treatment has revealed an important intratumoral bystander effect. A whole tumor can be eliminated when only a fraction of its tumor cells express TKh. We now report that the bystander effect not only acts within a tumor, but also between distant tumors. One TKh+ tumor was generated simultaneously with one or multiple TKh- tumors in different rat liver lobes such that there was no contact between the resulting tumors. Both the TKh+ and the TKh tumors regressed after GCV treatment and showed infiltration with macrophages and T lymphocytes. This distant bystander effect, which is likely immune mediated, should be of major importance for gene therapy of disseminated tumors. PMID- 9358029 TI - Long-term erythropoietin expression in rodents and non-human primates following intramuscular injection of a replication-defective adenoviral vector. AB - Erythropoietin (Epo)-responsive anemia is a debilitating complication of chronic renal failure and human immunodeficiency virus (HIV) infection that effects more than 150,000 Americans. Patients with Epo-responsive anemias are currently treated with repeated injections of recombinant human Epo. In the studies described in this report, we have examined the safety and efficacy of using a single intramuscular (i.m.) injection of replication-defective adenoviral vectors (RDAd) encoding Epo for the treatment of Epo-responsive anemias in both mice and non-human primates. Our results demonstrate that there is a threshold dose of virus (2.5-8 x 10(7) pfu/gram of body weight) which is required to obtain long term Epo expression and polycythemia in both species. A single i.m. injection of mice with 10(9) pfu of an RDAd encoding murine Epo (AdmEpo) resulted in elevations in hematocrits from control values of 49 +/- 0.9% to treated values of 81 +/- 3%, which were stable for more than 1 year. Similarly, a single i.m. injection of a monkey with 4 x 10(11) pfu of an RDAd-encoding simian Epo (AdsEpo) resulted in elevations of hematocrits from control levels of 40% to treated levels of > or =70%, which were stable for 84 days. Intramuscular injection of monkeys with AdsEpo appeared to be safe in that we did not detect abnormalities in chest X-rays, serum chemistries, hematologic, or clotting profiles (apart from elevated hematocrits) or organ histologies during the 84-day time course of the experiment. Taken together, these results suggest the feasibility of using i.m. injection of RDAd for the treatment of Epo-responsive anemias in humans. PMID- 9358032 TI - Bystander effects of different enzyme-prodrug systems for cancer gene therapy depend on different pathways for intercellular transfer of toxic metabolites, a factor that will govern clinical choice of appropriate regimes. AB - Transfer of suicide genes into tumor cells renders them sensitive to cytotoxic effects of specific prodrugs. We show here that both the herpes simplex virus thymidine kinase/ganciclovir (tk/GCV) and thymidine phophorylase/5'-deoxy-5 fluorouridine (tp/DFUR) suicide gene systems can induce cell death in tumor cells. Additionally in mixed cultures of cells with and without the suicide gene, death occurred in both cell types, indicative of a bystander effect. We demonstrate, in human and rodent cell lines, that the tk/GCV bystander effect requires gap junctional intercellular communication (GJIC). Where cultures lack GJIC, no bystander effect was observed. In communicating cultures, no correlation between level of GJIC and bystander effect was seen and this was due to high levels of tk activity. Additionally, we demonstrate that transfer of toxic metabolites from tk+ to tk- cells occurs within 2 hr of GCV application and, as no apoptosis could be detected until after this time, apoptosis is the result, not the cause, of the tk/GCV bystander effect. In the tp/DFUR system, a medium mediated bystander effect, independent of GJIC and apoptosis, was observed. We demonstrated that combining tk/GCV and tp/DFUR suicide gene systems in culture was more effective than either therapy alone. PMID- 9358031 TI - Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene. AB - Human hematopoietic stem cells remain one of the most promising target cells for gene therapeutic approaches to treat malignant and nonmalignant diseases. To rapidly characterize transduced cells and to isolate these from residual nontransduced, but biologically equivalent, cells, we have used a Moloney murine leukemia virus (Mo-MuLV)-based retroviral vector containing the intracytoplasmatically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene. Supernatant transduction of CD34+ cells (mean purity 97%) in fibronectin-coated tissue culture flasks resulted in 5.5-45% (mean 26%) transduced cells expressing deltaLNGFR (LNGFR+ cells). After transduction, more than 65% of the transduced cells remained CD34+. Compared with control (mock and nontransduced) CD34+ cells, transduction did not decrease the cloning efficiency of CD34+ cells. Immunomagnetic selection of the transduced cells with a monoclonal anti-LNGFR antibody resulted in >90% LNGFR+ cells. Further phenotypic characterization of these highly enriched LNGFR+ cells indicated that the majority co-expressed the CD34 and CD38 antigens. These results show that transduced cells expressing an ectopic cell-surface protein can be rapidly and conveniently quantitated and characterized by fluorescence-activated cell sorting (FACS) analysis and fast and efficiently enriched by immunoadhesion using magnetic beads. The use of cell-surface reporters should facilitate optimization of methods of gene transfer into more primitive hematopoietic progenitors. PMID- 9358033 TI - Organization, generation and replication of amphimeric genomes: a review. AB - Genomes comprising a pair of separated inverted repeats and called 'amphimers' are reviewed. Amphimeric genomes are observed in a large variety of different organisms, ranging from archaebacteria to mammals. The widespread existence of amphimeric genomes in nature could be due to their particular dynamic structure. Amphimeric genomes containing long inverted segments may provide the only form in which a duplicated segment is stably retained in genomes. Amphimers are often found in amplified subgenomes, indicating that they could promote a special mechanism of DNA replication and amplification. The possible mechanisms of generation, isomerization and replication/amplification of different types of amphimeric genomes are discussed. The study of amphimeric mitochondrial petite genomes of yeast could be a good model system for the study of the role of inverted repeat sequences in genome dynamics. PMID- 9358034 TI - Identification of a novel Escherichia coli gene whose expression is dependent on the flagellum-specific sigma factor, FliA, but dispensable for motility development. AB - FliA is an alternative sigma factor specific for class 3 flagellar operons. Using a promoter-probe vector, we randomly cloned Escherichia coli DNA fragments, which showed FliA-dependent promoter activities. Among the DNA fragments cloned, one was found to be derived from a non-flagellar region. Hybridization analysis with the Kohara E. coli library indicated that this DNA fragment is located at around 35.4 min on the E. coli chromosome where no flagellar gene has been reported yet. DNA sequence analysis revealed that it contains an FliA-dependent promoter-like sequence followed by an open reading frame (ORF) that can encode a 110-amino-acid protein. A rho-independent terminator-like sequence follows this ORF. This putative gene was named flxA. A gene disruptant was constructed by inserting the kan gene cassette into the flxA gene on the chromosome. This mutant was found to be actively motile, suggesting that this gene is unlikely to be involved in the motility phenotype of E. coli. PMID- 9358035 TI - Two vectors for the insertion of mammalian selectable genes into yeast artificial chromosome cloned DNA. AB - The introduction of cloned DNA into mammalian cells allows functional testing of genes contained on the fragments. In many cases, the exogenous DNA introduced into mammalian cells requires selectable genes that mark the presence of input DNA. Two new vectors, carrying mammalian selectable markers encoding for either neomycin-resistance (neo) or histidinol-resistance (hol), have been constructed for targeted integration to specific single-copy sites within yeast artificial chromosome (YAC) insert DNA. The integration cassettes comprise a single selectable yeast gene adjacent to a mammalian selectable gene, either LEU2 with neo or HIS3 with hol. Modification of the YAC occurs in yeast by transfection with linear DNA containing YAC-specific, unique, recombinogenic ends, thereby ensuring co-integration of the markers. Analysis of modified YACs confirms that both vectors correctly integrate into the targeted unique sites. The precise localization of selectable marker genes in the cloned DNA ensures the integrity of the genomic fragments during functional testing. Placement of mammalian selectable markers within the YAC insert DNA should allow for YAC-based gene targeting experiments in a variety of mammalian cell lines. PMID- 9358036 TI - A leech homolog of twist: evidence for its inheritance as a maternal mRNA. AB - In the development of leeches such as Helobdella robusta, mesodermal and ectodermal fates segregate to cells DM and DNOPQ, respectively, at fourth cleavage. As one step in identifying genes that may act in mesoderm determination, we have cloned the H. robusta homolog to the Drosophila gene twist. This homolog, designated Hro-twi, exhibits high (> 90%) amino acid identity with other twist-class genes within its basic-helix loop-helix (b-HLH) DNA binding motif and dimerization domain. Like twist, Hro-twi contains CAX-rich stretches: three stretches 5' to the b-HLH and one located 3' of the b-HLH motif. RT-PCR analysis suggests that Hro-twi is present throughout development, beginning as a maternal transcript in the oocyte. PMID- 9358037 TI - Human syntaxin 7: a Pep12p/Vps6p homologue implicated in vesicle trafficking to lysosomes. AB - The movement of hydrolases and other proteins to lysosomes is accomplished by vesicle trafficking. Specific vesicles are targeted from the trans-Golgi network via a prelysosomal compartment to lysosomes. The specificity of vesicle transport is thought to occur through the interaction of vesicle proteins with receptors on a particular target membrane. The syntaxins are a family of transmembrane proteins that have been implicated as vesicle receptors involved in vesicle docking and fusion. Syntaxins 1-4 are localized to the plasma membrane, and in particular, syntaxin 1a mediates synaptic vesicle docking in the nerve terminal. Syntaxins 5 and 6 have been localized to cis-Golgi and trans-Golgi network compartments, respectively. We now report the identification of syntaxin 7 from a human brain cDNA library. The syntaxin 7 gene is localized to human chromosome 6. By Northern analysis, the syntaxin RNA was found to be broadly distributed. Based on its homology to yeast and plant vacuolar syntaxins, we propose that syntaxin 7 has a role in vesicle trafficking between the Golgi complex and lysosomes. In vitro binding studies reveal that syntaxin 7 binds alphaSNAP, a key regulator of transport vesicle fusion at multiple stages of the secretory pathway. PMID- 9358038 TI - Sequence of the ponA gene and characterization of the penicillin-binding protein 1A of Pseudomonas aeruginosa PAO1. AB - The nucleotide sequence of the ponA gene encoding the high molecular-mass penicillin-binding protein 1A (PBP1A) of Pseudomonas aeruginosa (Pa) PAO1 was determined and characterized. The predicted PBP1A protein of 822 amino acids (aa) has a calculated molecular mass of 91.2 kDa corresponding to the size of the protein expressed in vitro and in vivo. A penicillin-binding (PB) assay showed that the Pa ponA gene product covalently binds penicillin. The deduced PBP1A aa sequence has features typical of class-A high-molecular-mass PBPs: a highly hydrophobic N-terminus portion containing a potential transmembrane segment which might anchor the protein to the cytoplasmic membrane; an N-terminal module with the conserved boxes 1 (E86D(DN)F(AN)H(Y)G), 2 (G117GS(T)I(TM)Q), 3 (R139K(IN)E(ILL)AL) and 4 (R221R(NW)IL); a PB module with the conserved boxes 5 (S461SFK), (S520RN) and (K695TG); an internal extension at aa 297-407 between the N-terminal and PB modules; and a C-extension at the end of the PB module at aa 742 to 822. The highest percentage of similarity (62.8%) was found with the class A high-molecular-mass PBP1A of Escherichia coli (Ec) and Haemophilus influenzae. The observed extensive homology in the modular design of the Pa PBP1A with the bifunctional Ec PBP1A suggests structural and functional relationships between these proteins and refutes the proposed correspondence between Pa PBP1A and Ec PBP1B. PMID- 9358039 TI - Structure and sequence of the bovine butyrophilin gene. AB - The complete sequence of the bovine butyrophilin gene (BTN) is described and compared with the mouse gene (Btn). Both genes contain seven exons separated by six introns, and the organisation of exons is closely associated with structural domains of the protein. Individual exons of BTN and Btn are 68-87% similar in sequence. There are no canonical TATA or CCAAT boxes associated with the transcription initiation sites in the genes of either species. However, a number of potential binding sites for transcription factors were identified in the 5' flanking DNA, some of which may function in regulating expression of the gene in mammary tissue. Conservation of a 110-bp region in the promoters of BTN and Btn may have some functional significance. Cloning and sequencing of BTN provides an additional mammary-specific gene promoter that may be used for driving the expression of transgenes in the lactating mammary gland, and for determining the basis for tissue-specific gene expression. In addition, the sequence of BTN may be used to map intragenic polymorphisms and identify quantitative trait loci in commercial livestock. PMID- 9358040 TI - Conservation of Y chromosome-specific sequences immediately 5' to the testis determining gene in primates. AB - Sex is determined in mammals by the SRY gene, which is located on the non recombining region of the Y chromosome. Although the presence of mutations in SRY associated with male to female sex reversal clearly indicates that this gene is essential for testis formation, little is known concerning other genes in this process or how the expression of SRY itself is controlled. This is mainly due to the absence of an appropriate in vitro cellular model. Previous studies have indicated that SRY coding sequences are undergoing rapid evolution in mammals. In this study, we cloned and compared a Y chromosome-specific region immediately 5' to the SRY open reading frame in several primate species. The divergence of the region 5' to SRY between primates was found to be comparable with that described for autosomal sequences. An alignment of sequences within the primate lineage, together with sequences from the cow and pig, revealed the presence of several highly conserved motifs. These domains may have a function in the control of SRY expression during fetal development. PMID- 9358041 TI - pRIBOTEX expression vector: a pTEX derivative for a rapid selection of Trypanosoma cruzi transfectants. AB - To improve the selection phenotype of the expression plasmid pTEX, a Trypanosoma cruzi rDNA (DNA coding for rRNA) gene spacer fragment (806 bp) containing a mapped transcription start point (tsp) was cloned in the vectors pTEX and pTEX cat, generating the plasmids pRIBOTEX and pRIBOTEX-cat. T. cruzi cultures transiently transfected with pRIBOTEX-cat expressed a chloramphenicol (Cm) acetyltransferase (CAT) activity 16,000-fold greater than the activity observed with the parental vector pTEX-cat. Moreover, T. cruzi cells transformed with pRIBOTEX and pRIBOTEX-cat exhibited logarithmic growth in the presence of Geneticin (G418) 2 weeks earlier than that observed with controls transformed with pTEX. The plasmid copy number in stably transformed trypanosomes was about 50-times higher in cultures transformed with pTEX-cat than in cells transformed with pRIBOTEX or pRIBOTEX-cat. However, the neo RNA steady-state level and the CAT activity observed among the stably transfected cultures showed only modest differences. Finally, it was found that the pRIBOTEX vector was not episomally maintained as pTEX, but integrated into a chromosome indistinguishable from the one encoding rRNA. These features make pRIBOTEX a useful tool for transfection and rapid expression of genes in T. cruzi. PMID- 9358042 TI - Cloning and characterization of the uvrD gene from an extremely thermophilic bacterium, Thermus thermophilus HB8. AB - The uvrD gene encodes a DNA helicase which plays an important role in prokaryotic nucleotide (nt) excision repair, mismatch repair and DNA replication. A cosmid based genomic DNA library for Thermus thermophilus (Tt) HB8 was constructed, and this was screened by Southern hybridization using a uvrD fragment amplified by PCR as the probe. The nt sequence of cloned Tt uvrD was then determined. Characteristic helicase motifs, made up of seven elements, were all conserved in the amino acid (aa) sequence of Tt UvrD. The aa sequence showed 41% homology with that of Escherichia coli (Ec). In the aa composition of Tt UvrD, the number of Asn, Gln, Met and Cys residues was decreased, and the number of Pro residues was increased. The distribution of Pro residues and recent data on X-ray crystallographic structure suggested the importance of the structural dynamics of the protein. These changes are thought to stabilize the native protein conformation against heat denaturation. Tt uvrD complemented the UV sensitivity of a Ec uvrD mutant. Thus, the thermophilic bacterium has a UvrD helicase, whose function is common to Ec UvrD. PMID- 9358043 TI - The hsp60 gene of the human pathogenic fungus Coccidioides immitis encodes a T cell reactive protein. AB - A heat shock protein-encoding gene (hsp60) from the human respiratory fungal pathogen, Coccidioides immitis (Ci), was cloned, sequenced, chromosome-mapped, expressed and immunolocalized in parasitic cells. Both the genomic and cDNA sequences are presented. The transcription start point and poly (A) addition site were confirmed. The hsp60 gene contains two introns and a 1782-bp ORF which translates a 594-amino acid (aa) protein of 62.4 kDa and pI of 5.6. The translated protein revealed two potential N-glycosylation sites. The deduced HSP60 showed 78-83% aa sequence similarity to reported fungal HSP60 proteins. The hsp60 gene was mapped to chromosome III of Ci and was shown to be a single copy gene by Southern and Northern hybridization. Expression of a 1737-bp cDNA fragment of the hsp60 gene in E. coli resulted in production of a recombinant protein. Amino acid sequence analysis of the recombinant protein confirmed that it was encoded by the Ci hsp60 gene. Antiserum raised in mice against the isolated recombinant protein immunolocalized HSP60 in the cytoplasm and wall of parasitic cells of Ci. The recombinant HSP60 was used to immunize BALB/c mice and was shown to induce proliferation of T cells isolated from lymph nodes of these animals. The hsp60 gene of Ci is the first reported heat-shock protein gene of this human pathogen. PMID- 9358044 TI - The sequences of hypF, hypC and hypD complete the hyp gene cluster required for hydrogenase activity in Bradyrhizobium japonicum. AB - A region of DNA 6 kb downstream of the hydrogenase (H2ase) structural genes and directly downstream of the hypB gene of Bradyrhizobium japonicum was shown by mutational analysis to be necessary for H2ase synthesis. Sequencing of this region revealed two complete open reading frames, and the 5' fragment of a third ORF. They encode proteins with homologies to the HypF, HypC and the N-terminus of HypD from other H2ase-containing organisms. The hypF of B. japonicum encodes a 753-aa protein with a predicted molecular mass of 80.3 kDa that contains the two zinc-finger motifs characteristic of other HypF proteins. The hypC encodes a 85 aa protein with a predicted molecular mass of 8.4 kDa. The 5' portion of hypD, which encodes the first 35 aa, upon combining with the previously reported C terminus of HypD, designated HypD' (Van Soom et al. (1993) Mol. Gen. Genet. 239, 235-240) encodes a protein with a predicted molecular mass of 42.4 kDa. Complementation studies on a H2 uptake defective strain of B. japonicum containing a polar mutation in the hyp operon revealed that the products of the hyp F, C, D, E genes are required for H2ase production. Evidence is also presented that the hyp genes are co-transcribed from a large operon together with the downstream genes hupGHIJK, making a polycistronic message of 11 genes. PMID- 9358045 TI - A member of the TGF-beta receptor gene family in the parasitic nematode Brugia pahangi. AB - The full length cDNA sequence of a Type I transforming growth factor-beta (TGF beta) receptor has been isolated from the filarial parasitic nematode Brugia pahangi. This new gene, designated Bp-trk-1, encodes a predicted 645 amino acid sequence with an N-terminal hydrophobic stretch which may act as a signal peptide. The extracellular portion (residues 15-187) is cysteine-rich and has three potential N-glycosylation sites. At positions 250-255 the protein contains the glycine-serine rich motif characteristic of Type I receptors. The closest homologue is a Caenorhabditis elegans gene (Q09488) in cosmid C32D5.2 which shares 67% amino acid identity with Bp-trk-1 in the most conserved kinase domain (aa 259-482). Other type I receptors such as C. elegans daf-1 and Drosophila tkv show 38-53% identity in the same region. Some residues conserved in Drosophila and vertebrates are not present in the B. pahangi sequence. RT-PCR amplification has been used to show that the transcript is expressed in the three main stages of the B. pahangi life cycle: microfilariae, infective larvae and adults. The ligand remains unknown at this time but is likely to be most similar to that for C. elegans Q09488. PMID- 9358046 TI - Identification and characterization of the thiamine transporter gene of Saccharomyces cerevisiae. AB - A positive selection scheme is described that selects for thiamine transporter clones. The scheme is based on the rescue of lethality, under non-permissive conditions, of Saccharomyces cerevisiae strains that are conditional for thiamine biosynthesis and are defective in thiamine transport. Transport defective strains were generated by selection for resistance to the lethal thiamine analog, pyrithiamine. Pyrithiamine resistance was shown to be a recessive, single gene trait that resulted from the mutation of the thiamine transporter gene, as suggested by previous work. Conditional thiamine biosynthesis was generated by cloning THI4, a thiamine biosynthetic gene, into a URA3 containing plasmid and transforming a strain disrupted in THI4. Thus, plating on 5-fluoroorotic acid causes the loss of thiamine synthesis ability. The gene for the yeast thiamine transporter, THI7, was cloned using this scheme. The predicted 598 amino acid transporter is a member of the major facilitator superfamily of transporters and thus possesses 12 transmembrane spanning segments with amino and carboxy termini intracellularly located. Several alterations in the coding region were characterized that result in greatly reduced ability to transport thiamine. The level of transporter mRNA was found to be rapidly and dramatically reduced by the addition of thiamine to the growth medium. PMID- 9358047 TI - Exon/intron structure of the human transferrin receptor gene. AB - A PCR-based intron jumping strategy has been utilized to investigate the exon/intron structure of the human transferrin receptor gene and determine the sequences of exon/intron junctions. There are 18 exons and introns 5' to a large exon encoding the last translated segment and a sizable 3' untranslated segment. All of the translated segments are encoded by exons 2-19. The tight turn motif, which is critical to the process of endocytosis, is encoded by exon 3. Based on recent studies of human/chicken receptor chimeras, it appears that the residues most likely to be involved in transferrin binding are encoded by exons 17-19. Exon 12 exhibits the greatest degree of homology with the gene for the prostate specific membrane antigen. A polymorphism has been tentatively identified at nucleotide position 519 in exon 4; the presence of either adenine or guanine should result in either serine or glycine, respectively, at position 142 of the amino acid sequence. This analysis of genomic structure will permit further detailed studies of the regulation, expression and evolution of the human transferrin receptor gene. PMID- 9358048 TI - Direct cloning of the Brassica S locus by using a P1-derived artificial chromosome (PAC) vector. AB - Self-incompatibility of Brassica is regulated by the S locus, which contains several genes including SLG and SRK. We found that both SLG and SRK genes were located at an approx. 80-kb MluI fragment in an S9 haplotype of B. campestris. Therefore, we cloned this MluI fragment into a BssHII site of the P1-derived artificial chromosome (PAC) vector. The utility of the direct cloning method is discussed in this study. PMID- 9358049 TI - Drosophila DPP2C1, a novel member of the protein phosphatase 2C (PP2C) family. AB - We report the molecular cloning, chromosome mapping and developmental transcription pattern of a putative serine/threonine protein phosphatase 2C (PP2C), DPP2C1, from Drosophila melanogaster. The 6-kb transcript of this first Drosophila PP2C gene encodes a 1428-aa deduced protein. The DPP2C1 protein contains a approximately 330-aa PP2C-like catalytic domain flanked by extensive N and C-terminal sequences showing no similarities to other PP2Cs. The dpp2c1 gene maps to 4E1-2 on the X chromosome, 1.5 kb upstream of the ddlc1 gene. Northern blot analyses showed that dpp2c1 transcription is developmentally regulated, accumulating maximally during early (0-6 h) and late (12-24 h) embryogensis. The presented molecular characterisation provides the basis for a genetic dissection of DPP2C1 function. PMID- 9358050 TI - Cloning and sequence analysis of cDNAs encoding plant cytosolic malate dehydrogenase. AB - Here we report the first complete sequence of plant cytosolic malate dehydrogenase (EC 1.1.1.37). The phylogenetic relationships between malate dehydrogenases from different cell compartments are discussed. The constructed phylogenetic tree shows that cytosolic NAD-MDH and chloroplast NADP-MDH have evolved through gene duplication of the pre-existing nuclear gene. PMID- 9358051 TI - Construction and expression of a modular gene encoding bacteriophage T7 RNA polymerase. AB - A modular gene that encodes T7 RNA polymerase (T7 RNAP) and consists of cassettes delimited by unique restriction sites was constructed. The modular and wild-type genes of T7 RNAP were cloned into a vector designed to express His-tagged proteins. The modular and wild-type genes provided the same level of protein expression (i.e., T7 RNAP represented up to 30% of the total protein in Escherichia coli strain BL21). Purification of both proteins by immobilized metal ion affinity chromatography (IMAC) resulted in similar yields (700-800 microg of enzyme per 20 ml of culture) and purity (>95%) as indicated by Coomassie blue staining, Western blotting and the absence of detectable contaminating nuclease activities. Both proteins exhibited identical efficiency in transcription assays, and their specific activities (about 200 U/microg) were close to that of a commercial T7 RNAP preparation. The modular gene provides a useful tool for cassette directed mutagenesis of T7 RNAP. PMID- 9358052 TI - Bacteriophage B103: complete DNA sequence of its genome and relationship to other Bacillus phages. AB - The genome of Bacillus subtilis bacteriophage B103 consists of double-stranded linear DNA 18,630 bp long. The DNA was sequenced, and the sequence was compared with DNA sequences of closely related phages, namely the members of the phage phi29 family. Among them, phage Nf was shown to be the most closely related to B103. Comparisons of several open reading frames (ORFs) among the family members helped to identify genes 1 and 5. A cluster of ORFs between genes 16 and 17 contains two ORFs with partial homology with two phi29 ORFs located in the same region. There are three more ORFs in this region of B103 with good ribosome binding sites (RBS) and optimal codon usage that are not homologous to any of the phi29 ORFs. The function of these five ORFs remains unexplained. It was shown that major promoters characterized in phi29 are retained in B103. Where many substitutions occur in the vicinity of a promoter, at least the -10 and -35 boxes are conserved. PMID- 9358053 TI - Upstream genomic sequence of the human connexin26 gene. AB - Human connexin 26 (Cx26) has been considered to be a candidate suppressor gene in mammary epithelial cells. To gain insight into the transcriptional regulation of this gene, we have cloned and sequenced the 5' portion of the gene, which extends 4.8 kb upstream from the ATG translation start site. The 3' end of the non-coding exon 1 (160 bp) is located at 3149 bp upstream from the 5' end of exon 2. Comparison between the human Cx26 gene and the mouse gene reveals a highly conserved promoter region with 81% homology. In addition to six GC boxes and two GT boxes, a TTAAAA box is located at -24 to -19 bp upstream of the transcription start point. Analogous to the mouse beta-casein gene, the promoter region of the human Cx26 gene also contains a YY1-like binding site and a consensus mammary gland factor binding site. PMID- 9358054 TI - Tissue-specific alternative RNA splicing of rat vesicle-associated membrane protein-1 (VAMP-1). AB - The vesicle-associated membrane protein (VAMP) family is essential to vesicle mediated protein transport. Three mammalian isoforms, VAMP-1, VAMP-2, and cellubrevin, play a role in protein transport to the plasma membrane. In this study, we describe a new rat VAMP-1 isoform produced by alternative pre-mRNA splicing. Only one VAMP-1 isoform dominates in each tissue. Analysis of the nucleotide sequence for the newly discovered isoform, VAMP-1b, reveals that its expression is determined by whether an intron is retained or removed. The predicted amino acid sequences for the VAMP-1 isoforms differ at the carboxy terminal end of the protein. A similar process has been described for VAMPs in Drosophila melanogaster and suggests a conserved function for the carboxy terminal domain that can be modulated. PMID- 9358056 TI - Cloning and characterisation of a novel P-glycoprotein homologue from barley. AB - P-glycoproteins are members of a large superfamily of transport proteins (the 'traffic ATPases') that utilize ATP to translocate a wide range of substrates across biological membranes. Using a PCR-based approach, and degenerate oligonucleotides corresponding to conserved motifs, two 300-bp cDNA fragments (pBMDR1 and pBMDR2) with a significant sequence similarity to mammalian P glycoproteins were amplified from barley (Hordeum vulgare) root poly A+ RNA and used as probes to screen a barley root cDNA library. A single full-length clone pHVMDR2 coding for a polypeptide of 1232 residues (c. 134 kDa) was isolated. Comparison of this barley sequence with Arabidopsis ATPGP1 and human MDR1 and MDR3 P-glycoprotein sequences showed that the barley cDNA has 44%, 37% and 38% amino acid (aa) identity, respectively, with these sequences, and conserved structural features. RNase protection analysis showed that HVMDR2 mRNA is expressed at low levels in both barley roots and leaves. Southern blot analyses indicated that there is a small multigene family related to P-glycoproteins in barley. Possible functions for these barley P-glycoproteins are discussed. PMID- 9358055 TI - Ku80 gene expression is Sp1-dependent and sensitive to CpG methylation within a novel cis element. AB - The Ku70/80 complex, known as Ku, constitutes the DNA end binding component of the DNA-dependent protein kinase (DNA-PK). We have characterized the promoter region of the mouse and human Ku80 genes to delineate transcriptional elements necessary for basal gene expression and proliferation-dependent regulation. Consensus Sp1 recognition elements were identified in both promoters, and were determined to be essential for basal expression. We further identified a near perfect palindrome of 21 base pairs located immediately 5' to one Sp1 element. This sequence was present once within the mouse Ku80 promoter and seven times, in a head-to-tail tandem array, within the human Ku80 promoter. This sequence possessed homology with a methylation-sensitive promoter element, Enh2, present in the LTR of mouse intractisternal A-particles. Promoter deletion studies and expression analysis of in-vitro methylated reporter gene constructs provided strong evidence that, in vivo, this repeat sequence regulates Ku80 gene expression in cis, through a mechanism involving CpG methylation. Evidence is also presented, suggesting that Ku is directly involved in this regulatory process. PMID- 9358057 TI - Characterization of the gntT gene encoding a high-affinity gluconate permease in Escherichia coli. AB - We characterized the gntT gene encoding a high-affinity gluconate permease of Escherichia coli K-12. Primer extension and lacZ-operon fusion analyses revealed that gntT has one strong and two weak promoters, all of which are regulated positively by cAMP-CRP and negatively by GntR. The weak promoters became constitutive when separated from the upstream region including the strong promoter that overlaps a putative GntR-binding sequence. Gluconate-specific uptake activity was observed with cells harboring the gntT plasmid clone, which was enhanced by the presence of gntK encoding gluconate kinase. PMID- 9358058 TI - Analysis of bovine selenoprotein P-like protein gene and availability of metal responsive element (MRE) located in its promoter. AB - Selenoprotein P-like protein, similar to selenoprotein P, uses multiple TGAs for incorporation of selenocysteines but not as stop codons. It is also characterized by having a His-Pro-rich domain and a regionally differential expression pattern. Hence, in addition to selenium metabolism, this protein is considered to have a developmental function. In the present study, the structure of the selenoprotein P-like protein gene was analyzed. The gene consisted of five exons, and the 5' flanking region contained a TATA box, TCF-1-CS, bHLH-CS, gamma-IRE-CS, c-Myb-CS, C/EBP-CS, HNF-5-CS, MRE2-CS, etc. The presence of motifs like TCF-1-CS, c-Myb-CS, etc. supports the suggestion that this protein is involved in cellular maturation. Since the presence of MRE2-CS suggests that this protein is related to the antidote effect of selenium against heavy metal intoxication, the availability of this motif was examined using bovine kidney cell lines, CKT-1 and MDBK. Metallothionein mRNA markedly increased 6 h after administration of 10(-6) M CdCl2 and ZnCl2 in both cell lines. No significant alteration was observed in selenoprotein P-like protein mRNA, whereas its basal expression was high, indicating that this protein is constitutively expressed. Thus, it is still possible that this protein acts as an antidote, even though it is not inducible by heavy metals. PMID- 9358059 TI - Identification of adjacent genes encoding the major catalase and a bacterioferritin from the plant-beneficial bacterium Pseudomonas putida. AB - The catA gene, encoding the major CatA from a root-colonizing isolate Pseudomonas putida (Pp), was cloned by complementation into a catalase (Cat)-deficient Escherichia coli (Ec) strain UM2. The ORF for catA consisted of 479 aa with a higher degree of identity with typical Cat from eukaryotes than prokaryotes. Chromosomal homologous exchange with a mutant gene bearing an insertion of a luxAB-npt cassette into the SfiI site of catA generated a CatA-deficient Pp isolate. This mutant and another mutant, J1M, derived by EMS mutagenesis, were highly sensitive to hydrogen peroxide. CatA activity and resistance to hydrogen peroxide were restored in both mutants by catA. Adjacent to the 3' end of catA was a potential ORF of 462 nt that had high identitity with other bfr genes that encode iron-storage proteins. Northern analysis of the bfr gene from Pp revealed a transcript of approximately 500 nt. CatA and bfr probes hybridized to the same size restriction fragments in genomic DNAs from other root-colonizing and plant pathogenic pseudomonads. Thus, the genes for an iron-storage protein and the heme containing Cat appear to be conserved in adjacent loci in certain pseudomonads. PMID- 9358060 TI - Cloning and sequence analysis of the gene (eprA1) encoding an extracellular protease from Aeromonas hydrophila. AB - A gene (eprA1) encoding the extracellular protease of Aeromonas hydrophila AH1 has been cloned and sequenced. Nucleotide sequence analysis of eprA1 predicted a single open reading frame (ORF) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. When the eprA1 gene was expressed in minicells, one major band of approx. 37 kDa was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kDa determined by SDS-PAGE analysis of the minicells. The deduced C-terminal aa region (Arg-290 to Gly-313) showed sequence homology to partial C-terminal sequences of other zinc metalloproteases including Penicillium citrinum metalloprotease (PlnC), Aspergillus oryzae metalloprotease (NpII), Aspergillus flavus metalloprotease (MepA), and Aspergillus fumigatus metalloprotease (Mep20), particularly with respect to zinc-binding residues. PMID- 9358061 TI - Isolation of the Staphylococcus aureus hemCDBL gene cluster coding for early steps in heme biosynthesis. AB - We have recently reported [Kafala, B., Sasarman, A., 1994. Can. J. Microbiol. 40, 651 657] the cloning and sequencing of the Staphylococcus aureus hemB gene. This gene purportedly encodes the delta-aminolevulinic acid dehydratase of the heme pathway. In this present communication, we report the sequences and identities of three putative hem genes. Two of these genes are located immediately upstream from hemB. Complementation analysis of Escherichia coli and Salmonella typhimurium hemC and hemD mutants and the comparison of the Sa nucleotide sequences with those of Bacillus subtilis and Ec showed that these two open reading frames, ORF1 and ORF2, are likely to be the hemC gene coding for porphobilinogen deaminase and the hemD gene coding for uroporphyrinogen III synthase, respectively. The third hem gene, hemL, is located immediately downstream of hemB, and encodes glutamate 1-semialdehyde 2,1-aminotransferase. Sequencing of the region which extends past hemL indicates that no further hem genes are located downstream of hemL. In Sa, hemC, hemD, hemB and hemL are proposed to constitute a hem cluster encoding enzymes required for the synthesis of uroporphyrinogen III from glutamate 1-semialdehyde (GSA). PMID- 9358063 TI - Positioning of 72 potentially full size LTRs of human endogenous retroviruses HERV-K on the human chromosome 19 map. Occurrences of the LTRs in human gene sites. AB - Seventy-two near full size long terminal repeats (LTRs) of human endogenous retrovirus of K-family (HERV-K) have been precisely located on the metric map of human chromosome 19. The LTR-related sequences were identified and assigned to cosmids by hybridization with two independent chromosome 19 specific cDNA clones corresponding to different parts of U3 region of LTR of HERV-K. The presence of full-size LTR sequences in a cosmid was further verified by PCR assay with a pair of primers complementary to the termini of the LTR. Coincidences of the LTR and the known genes positions are discussed. PMID- 9358062 TI - Structure, organization and putative function of the genes identified within a 23.9-kb fragment from Arabidopsis thaliana chromosome IV. AB - In the framework of the complete genome sequencing programme of the crucifer Arabidopsis thaliana, a 23.9-kb fragment from the long arm of chromosome IV has been analysed. This paper presents a methodological approach, integrating computerized predictions, database screening, the sequencing of cognate cDNAs and a PCR-based detection of expression that allows the accumulation of an important amount of information from an anonymous sequence. This work revealed the organization of novel genes and the vestige of a copia-like retrotransposon. The gene AtRH1 encodes the first member of a new subfamily of the plant DEAD box RNA helicases. A recurrent and complete search of dbEST has been used to evaluate the number of different RNA helicases expressed in A. thaliana. On the 18 discriminated members of the family, only a small number seems to be expressed at a relatively high level. The putative gene AtTS1 encodes a novel terpene synthase in A. thaliana, and the genes G14587-5 and G14587-6 encode unknown proteins. This study illustrates most of the situations that could be encountered during the analysis of an anonymous sequence from A. thaliana. PMID- 9358064 TI - Cloning, nucleotide sequence and characterization of a full-length cDNA encoding the myosin heavy chain from adult chicken pectoralis major muscle. AB - Four cDNA clones, encoding the chicken adult sarcomeric MyHC, have been isolated from a pectoralis major muscle cDNA library using gene-specific DNA probes. These clones were sequenced and then subcloned into a full-length, 6-kb, chicken adult sarcomeric MyHC cDNA. The entire cDNA consists of 5873 nucleotides with 19 bp 5' untranslated region and 34 bp 3'-untranslated region. The complete cDNA encodes a 1939-aa polypeptide whose molecular weight is 223 kDa. The calculated isoelectric point of this protein is approximately 5.7. Analysis of the deduced amino acid sequence and comparison with a previously published amino-acid sequence of the same MyHC isoform reveals that six amino acid residues are different. Hydrophilicity analysis of this adult MyHC amino-acid sequence shows a similar pattern as the embryonic MyHC. A recombinant baculovirus, carrying this full length adult MyHC cDNA, has also been generated and expressed in the Sf9 insect cell line. A approximately 220-kDa recombinant MyHC was synthesized and reacted specifically with chicken adult MyHC monoclonal antibodies. PMID- 9358065 TI - The Azotobacter vinelandii alg8 and alg44 genes are essential for alginate synthesis and can be transcribed from an algD-independent promoter. AB - A 2.8-kb DNA region, located immediately downstream of algD, contains the A. vinelandii alg8 and alg44 genes, whose sequences are highly homologous to those of the corresponding Pseudomonas aeruginosa genes. These genes occur on a transcript that does not include algD, and are transcribed from a promoter different from that transcribing algD; this is the fourth promoter described within the alginate biosynthetic gene cluster. alg8 and alg44 mutants were constructed and shown to be completely impaired in alginate production. Alg8 shares 28.20% identity and 38.09% similarity to Azorhizobium caulinodans NodC, a glycosyl transferase catalyzing the formation of beta-1,4 linkages. A topological model is predicted, which supports the idea of Alg8 being the polymerase responsible for alginate synthesis. PMID- 9358066 TI - A family of repetitive DNA sequences in Old World primates. AB - A dispersed family of repetitive DNA sequences that is amplified in Old World primates has been characterized. The sequences are present in about 250-350 copies in humans, found on all chromosomes, and some are at least 1 kb in size. Within the core repeat is a 178-bp region that is moderately-to-highly conserved. A representative sequence exhibited strong promoter activity when placed in front of a promoterless gene and transfected into human cells. This promoter activity has been localized to a 138-bp region of the repeat that is about 150 bp downstream of the 178-bp conserved region. Transcripts of the sequences were not detected in six human breast epithelial and teratocarcinoma cell lines. Based upon the work of Pavelitz et al. [Pavelitz, T., Rusche, L., Matera, A.G., Scharf, J.M., Weiner, A.M., 1995. EMBO J. 14, 169-177], the sequence appears to be related to the LTR of an HERV-K class human endogenous retrovirus. PMID- 9358067 TI - Sequencing analysis of prion genes from red deer and camel. AB - An abnormal isoform of the prion protein (PrP) appears to be the agent responsible for transmissible spongiform encephalopathies (TSE). The normal isoform of PrP is host-encoded and expressed in the central nervous system. The recent bovine spongiform encephalopathy (BSE) epidemic in the UK and the incidence of prion-related diseases in other animals could indicate that ruminants are highly susceptible to infection via ingestion of prion-contaminated food. Sequence analysis of PrP gene open reading frames from red deer and camel was carried out to investigate sequence variability of these genes among ruminants. PMID- 9358069 TI - Codon optimization for high-level expression of human erythropoietin (EPO) in mammalian cells. AB - Codon bias has been observed in many species. The usage of selective codons in a given gene is positively correlated with its expression efficiency. As an experimental approach to study codon-usage effects on heterologous gene expression in mammalian cells, we designed two human erythropoietin (EPO) genes, one in which native codons were systematically substituted with codons frequently found in highly expressed human genes and the other with codons prevalent in yeast genes. Relative performances of the re-engineered EPO genes were evaluated with various combinations of promoters and signal leader sequences. Under the comparable set of combinations, mature EPO gene with human high-frequency codons gave a considerably higher level of expression than that with yeast high frequency codons. However, the levels of EPO expression varied, depending on the alternate combinations. Since the promoters and the signal leader sequences that we used are known to be equally efficient in gene expression, we hypothesized that the varied expression levels were due to the linear sequence between the promoter and the coding gene sequence. To test this possibility, we designed the EPO gene with hybrid codon usage in which the 5'-proximal region of the EPO gene was synthesized with yeast-biased codons and the rest with human-biased codons. This codon-usage hybrid EPO gene substantially enhanced the level of EPO transcripts and proteins up to 2.9-fold and 13.8-fold, respectively, when compared to the level reached by the original counterpart. Our results suggest that the linear sequence between the promoter and the 5'-proximal region of a gene plays an important role in achieving high-level expression in mammalian cells. PMID- 9358068 TI - A 'core ATPase', Hsp70-like structure is conserved in human, rat, and C. elegans STCH proteins. AB - We have identified the rat and Caenorhabditis elegans homologues of a 'core ATPase'-encoding Hsp70-like gene, designated Stch. We observed that the human, rat, and C. elegans Stch genes have conserved a stop codon immediately distal to the sequence encoding the Hsp70 ATPase domain. This results in the functional equivalent of an N-terminal, proteolytically cleaved fragment of Hsc70/BiP. Each homologue contains a hydrophobic signal sequence, demonstrates striking identity within the Hsp70 ATPase domain, and retains a similar C-terminal sequence (STCH specific cluster III) that is unique among Hsp70 proteins and which truncates the peptide binding domain. In addition, we have identified an internal 35-aa region that is homologous to the minimal sequence of the Hip chaperone co-factor that is required for direct binding to the ATPase domain of Hsp70. Adjacent to this region, the rat and human STCH protein sequences diverge within a short internal 'insertion' sequence that interrupts the ATPase subdomain between the phosphate-2 and adenosine ATP-binding sites. We have also demonstrated that both human and rat Stch are constitutively produced and are induced by the calcium ionophore A23187, but not by heat shock. The recognition that the truncated 'core ATPase' structure of the STCH molecule is conserved in human, rat, and C. elegans tissues suggests an important role for this unique member of the membrane-bound Hsp70 family. PMID- 9358070 TI - Primary structure of Neurospora crassa gamma-tubulin. AB - We report the isolation and characterization of a cDNA-clone for gamma-tubulin from N. crassa. The deduced amino-acid sequence of the cDNA-clone most resembles that of gamma-tubulins from other fungi. Differential Southern blot analysis demonstrates that most probably a single gene represents gamma-tubulin. In addition, Northern blot analysis shows that most probably only one single mRNA encoding gamma-tubulin is transcribed in N. crassa. In a phylogenetic tree that includes all known gamma-tubulins, N. crassa gamma-tubulin is grouped, as expected, together with other fungal gamma-tubulins. PMID- 9358071 TI - Can insulin-dependent diabetes mellitus be understood at the molecular level through Na+/K+-adenosine triphosphatase? PMID- 9358072 TI - Recombinant human erythropoietin to assist autologous blood donation by patients with anemia. PMID- 9358073 TI - Matrix and adhesion molecules in kidney pathology: recent observations. AB - The purpose of this article is to review a set of recently obtained data concerning matrix and matrix adhesion molecules in renal disease. Our goal is not to cover the entire topic, but rather to focus on findings obtained with an experimental model for chronic lupus nephritis, evoked in mice by inducing graft versus-host disease (GVHD). The overall aim of these studies was to investigate the role of adhesion molecules as targets for autoantibodies, in the recruitment of inflammatory cells, and in the accumulation of matrix in kidney disorders. In addition, we set out to discover how matrix proteins in renal diseases differ from normal matrix molecules both quantitatively, in their increased frequency, and qualitatively, in their intramolecular structure. The advances in understanding and methodology described in this review imply a substantial capability for greater insight into the pathogenesis of kidney disease; for making better use of renal biopsies, such as in applying competitive reverse transcriptase-polymerase chain reaction (RT-PCR) in RNA analysis for matrix; and in developing more effective treatment strategies for patients with kidney disease. PMID- 9358074 TI - Polycythemia vera: a retrospective and reprise. AB - This article, by two of the late John H. Lawrence's fellows of the 1940s, traces the development of the knowledge of polycythemia vera from Vaquez, who wrote the first description of this disease, and Osler, who recognized it as "a new clinical entity," through John H. Lawrence and the use of 32P as a treatment for polycythemia vera, to the formation of French and Italian polycythemia study groups. In particular, the history of polycythemia vera after the Second World War, and its more recent history, can be traced through the development of an algorithm for evaluating an elevated hematocrit and the development of the first (O1) protocol of the Polycythemia Vera Study Group (PVSG), a randomized trial of the efficacy of 32P, chlorambucil, and phlebotomy for treating polycythemia vera. It was in 1948, only 9 years after the first use of 32P for treating polycythemia vera, that Byron Hall reported the occurrence of acute leukemia following this use of the isotope. This led to the formation of the PVSG. After completing enrollment of patients in the first protocol of the PVSG, an attempt to find a replacement for 32P as a myelosuppressive agent led to the testing of hydroxyurea as a putative non-leukemogenic drug for this purpose. However, the use of hydroxyurea for treating polycythemia vera is coming into question, as is the ability to maintain patients with phlebotomy alone. The PVSG as such no longer exists as an operational group; its files are maintained at the Mount Sinai School of Medicine in New York City. However, the French group created for the study of polycythemia vera has had a consensus conference, and the Italian group has developed a low-dose aspirin protocol for treating the disease. PMID- 9358075 TI - Modifications induced by insulin-dependent diabetes mellitus on human placental Na+/K+-adenosine triphosphatase. AB - The causes of the reduced activity of Na+/K+-adenosine triphosphatase (ATPase) in human diabetes are still the object of controversy. The aim of this work was to investigate the mechanisms of inhibition by means of the study of the Na+/K+ ATPase purified from human placenta. We purified Na+/K+-ATPase from term placentas of six healthy women and six age-matched women with insulin-dependent diabetes mellitus (IDDM) in good metabolic control. The enzymatic activity was reduced in both the microsomal fraction and the purified Na+/K+-ATPase obtained from diabetic women, whereas no difference was found in the number of active molecules determined by anthroyl ouabain binding. The Na+/K+-ATPase purified from women with IDDM did not show any modification in the ouabain affinity or changes in the physicochemical structure of the ouabain binding site investigated by dynamic fluorescence or alterations in lateral diffusion. The activation energy of the enzyme was increased, whereas the tryptophan accessibility of the enzyme was lower in women with IDDM. The fluidity of the lipid anulus of the enzyme was higher in women with IDDM than in control women, as suggested by fluorescence polarization of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene. The adenosine triphosphate-binding site, investigated by anisotropy decay studies of the fluorescent probe pyrene isothiocyanate, was modified in women with IDDM. It appears that the Na+/K+-ATPase of human placenta is altered in its disposition in IDDM. PMID- 9358076 TI - The erythropoietic response to erythropoietin in patients with rheumatoid arthritis. AB - We studied whether orthopedic surgical patients with rheumatoid arthritis (RA) can generate an erythropoietic response to either endogenous erythropoietin or to recombinant human erythropoietin (EPO) therapy to the same extent as patients without rheumatoid arthritis (non-RA). Seventy patients (10 RA, 60 non-RA) were entered into clinical trials of aggressive autologous blood donation before elective orthopedic surgery at one institution, randomized to receive EPO (600 U/kg, iv, 6 times over 3 weeks) or placebo. RA patients given EPO had red blood cell (RBC) production that was enhanced by 624 +/- 137 ml (mean +/- SD) as compared with 271 +/- 174 ml (p = 0.02) for RA patients given placebo treatment. Preoperative RBC volume expansion in 10 RA patients was 5.9 +/- 3.7 ml/kg as compared with 7.4 +/- 3.9 ml/kg for 60 non-RA patients (p = 0.13). RA patients can benefit to the same extent as non-RA patients from aggressive blood conservation programs that incorporate erythropoietin-modulated erythropoiesis. PMID- 9358077 TI - Gene expression of lipoprotein lipase in experimental nephrosis. AB - Nephrotic syndrome (NS) is commonly associated with marked hypertriglyceridemia, impaired triglyceride-laden lipoprotein clearance, and reduced peripheral tissue uptake of triglycerides from chylomicrons. Lipoprotein lipase (LPL) is the rate limiting step in triglyceride-rich lipoprotein metabolism. Earlier studies have demonstrated a marked reduction of plasma post-heparin lipolytic activity and LPL protein in NS. However, the effect of NS on gene expression of LPL has not been elucidated. We studied rats with puromycin aminonucleoside-induced NS and the placebo-injected control animals. Heart, soleus muscle, and fat body LPL activity, protein mass, and mRNA were measured and plasma lipid levels were quantitated. The NS group exhibited marked proteinuria, hypoalbuminemia, and hypertriglyceridemia. This was associated with significant reductions of LPL activity and immunodetectable protein in the heart, adipose tissue, and soleus muscle in the NS group. The reduction in LPL protein mass in the tissues tested was accompanied by a parallel reduction in LPL mRNA of the heart but not of either adipose tissue or skeletal muscle, suggesting translational or posttranslational modifications. A negative correlation was found between plasma triglyceride concentration and the LPL, activities of the tissues tested in the study population. Thus this study has revealed a significant down-regulation of tissue LPL protein in experimental NS. This phenomenon can, in part, account for hypertriglyceridemia, impaired catabolism of chylomicrons, and very low-density lipoprotein by peripheral tissues and decreased postheparin lipolytic activity in NS. PMID- 9358078 TI - Angiotensin-converting enzyme and neutral endopeptidase modulate smokeless tobacco-induced increase in macromolecular efflux from the oral mucosa in vivo. AB - Smokeless tobacco elicits plasma exudation from the oral mucosa that is mediated by bradykinin, and it decreases the activity of tissue angiotensin-converting enzyme (ACE), a peptidase that cleaves and inactivates bradykinin. However, the mechanisms regulating bradykinin-induced responses during exposure to smokeless tobacco are uncertain. The purpose of this study was to begin to address this issue by determining whether inhibitors of ACE and neutral endopeptidase (NEP), a membrane-bound peptidase widely distributed in the oral mucosa that also cleaves and inactivates bradykinin, potentiate a smokeless tobacco-induced increase in macromolecular efflux from the oral mucosa in vivo. Using intravital microscopy, we found that suffusion of an aqueous extract of smokeless tobacco elicited a significant concentration-dependent increase in fluorescein isothiocyanate labeled dextran (molecular mass 70 kd) leaky site formation in the hamster cheek pouch (p < 0.05). This response was significantly potentiated by captopril and lisinopril, two ACE inhibitors, and by phosphoramidon and thiorphan, two NEP inhibitors (p < 0.05). The effects of ACE and NEP inhibitors were additive. By contrast, a mixture of proteinase inhibitors consisting of leupeptin, Bestatin, and DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid had no significant effects on smokeless tobacco extract-induced responses. Overall, these data suggest that ACE and NEP each play a role in modulating a smokeless tobacco induced increase in macromolecular efflux from the in situ oral mucosa, in part by regulating local bradykinin catabolism. PMID- 9358079 TI - Monitoring platelet glycoprotein IIb/IIIa-fibrin interaction with tissue factor activated thromboelastography. AB - Computerized thromboelastography (TEG) was used to study platelet glycoprotein IIb/IIIa function, characterize the consequences of the interaction between polymerizing fibrin and activated platelets, and establish a quantitative assay of platelet function. The ability of platelets to augment the shear elastic modulus of blood clots was measured by TEG under conditions of maximal platelet activation during ex vivo clot formation accelerated by recombinant human tissue factor (TF). Under these conditions, platelets significantly enhance clot strength eightfold (relative to platelet-free fibrin clots). This effect, inhibited by cytochalasin D and c7E3 Fab, appears to be dependent on the transmission of platelet contractile force to fibrin, through glycoprotein IIb/IIIa receptors. The c7E3 Fab dose response of TF-TEG clot strength is identical to results with platelet aggregation induced by the thrombin receptor agonist peptide (50% inhibitory concentration (IC50 = 3.6 microg/ml); adenosine diphosphate-induced aggregation is easier to inhibit (IC50 = 1.2 microg/ml). The results obtained with this system are reproducible (coefficient of variation for clot strength 4%; intraclass correlation coefficient 0.96). As a clinical assay, TF-triggered computerized TEG is easy to perform at the bedside, provides on-line results within 30 minutes, and may offer advantages over conventional measures of platelet function. PMID- 9358081 TI - Technical and surgical aspects of continuous vascular access in freely moving small animals. AB - Many systems and techniques for continuous vascular access in small animals have been described. Problems with these systems have included (1) insufficient free movement, (2) sepsis, (3) high cost, (4) complicated construction, (4) thrombosis, and (5) dislocations of the intravenous catheter. The described operative techniques and a new experimental setup overcome these complications. The apparatus involves a swivel that is connected with an intravenously placed polyurethane catheter. A leather harness on the back of the animal is connected with the end of the swivel joint via a silicone tube in which the intravenous catheter runs to the swivel. The swivel, a modified conventional glass syringe, is positioned in ball bearings and a Johnson joint. The swivel, ball bearings, and Johnson joint are counterbalanced and can move up and down. When this system was used, the catheters functioned well for as long as 28 days, with a mean duration of 24.4 +/- 1.8 days (n = 420). Five catheter dislocations resulted from harness failure, and three dislocations were caused by animals twisting. All animals gained weight (3.53 +/- 0.37 gm/day (mean +/- SEM)). The rotary portion of the swivel and the Johnson joint secure stressless movement of the animal, avoiding twisting and dislocation of the catheter, which overcomes typical problems of existing methods. The low thrombogenicity of the polyurethane catheter also reduces complications. A further advantage is low cost, because prefabricated, reusable materials are used. PMID- 9358080 TI - A baboon model for hematologic studies of cardiopulmonary bypass. AB - Objective investigation of new inhibitors of blood protein or cellular systems that are activated during cardiopulmonary bypass (CPB) is impeded by the absence of a satisfactory animal model. Because most baboon hematologic proteins immunologically cross-react with those used for human assays, we developed a robust, reusable baboon model of CPB. Blood samples were obtained from adult baboons at six time intervals before, during, and after 60 minutes of partial CPB at 37 degrees C with peripheral cannulas. Both membrane (n = 7) and bubble oxygenators (n = 7) were investigated. We measured platelet and white blood cell counts; platelet response to adenosine diphosphate and release of beta thromboglobulin; fibrinopeptide A, prothrombin fragment F1.2, thrombin antithrombin complex, D-dimer, and plasmin-antiplasmin complex; activated complement (C3b/c and C4b/c); elastase-alpha1 proteinase inhibitor complex; and bleeding times. Adherent glycoprotein IIIa antigen in Triton X-100 washes of the perfusion circuit was also measured. Markers of baboon platelet, complement, and neutrophil activation and thrombosis significantly increased during CPB with bubble oxygenator systems but did not change appreciably in membrane oxygenator circuits. Markers of fibrinolysis, D-dimer, and plasmin-antiplasmin complex did not change with either oxygenator. The baboon model of CPB, when a bubble oxygenator is used, is a robust, reusable animal model for evaluating inhibitors of platelet, complement, and neutrophil activation and thrombosis during and after CPB. PMID- 9358082 TI - Interactions of recombinant hemoglobin (rHb1.1) and endotoxin in vivo: effects on systemic tumor necrosis factor and interleukin-6 levels in lethal and sublethal murine models of endotoxemia. AB - The effects of acellular hemoglobin-based oxygen carriers in preclinical models of sepsis and endotoxemia have been inconclusive with regard to outcomes reported for survival. In the present study, mice were infused with 1 gm/kg of recombinant human hemoglobin, rHb1.1, and the effects on mortality and systemic tumor necrosis factor (TNF) and interleukin-6 (IL-6) levels were determined by using both lethal and sublethal bolus endotoxin challenge. Pretreatment of mice with rHb1.1 and challenge with 20 mg/kg of lipopolysaccharide (LPS) at an LD100 resulted in a 100% mortality rate by 20 hours, whereas the same mortality rate with the vehicle or 5% albumin groups occurred at 50 hours. Mice challenged with lower LPS concentrations of 10 and 2.5 mg/kg, corresponding to LD15 and LD0, respectively, had 100% and 17% mortality rates in the rHb group and 17% and 0% mortality rates in the vehicle-treated animals. These doses of LPS resulted in maximal increases in systemic TNF, and there were only modest differences between the rHb and the vehicle groups at LPS challenge doses of 2.5 and 20 mg/kg, whereas no difference was observed at the 10 mg/kg concentration. At LPS concentrations below 10 microg/kg, the increases in circulating TNF were dose dependent and no differences were observed in serum TNF levels between the rHb1.1 and vehicle groups. In addition, there were generally no differences in IL-6 levels between the experimental groups, although at 10 mg/kg LPS, a twofold increase in plasma IL-6 levels over those in the controls was observed in the rHb1.1-treated animals. Infusion of rHb1.1 alone did not induce any increase in circulating IL-6 or TNF. These data demonstrate that endotoxin exacerbation, although apparent, was observed only at the highest doses of LPS and that at lower concentrations, there were no differences in the extent of cytokine elevation or in survival rate when rHb1.1-, albumin-, or vehicle-pretreated animals were compared. PMID- 9358083 TI - Glucose measurement in patients with diabetes mellitus with dermal interstitial fluid. AB - Although measurement of capillary blood glucose remains the standard method of self-monitoring for persons with diabetes mellitus, a less-invasive method of monitoring would be desirable. Measurement of dermal interstitial fluid glucose might meet this need. To test this possibility, plasma glucose, capillary blood glucose (current standard), and dermal interstitial fluid glucose were measured in 17 subjects with type I diabetes during a 5-hour pre- and postprandial period when plasma glucose was changing rapidly. The objective was to assess the ability of dermal interstitial fluid glucose to accurately predict plasma glucose over a wide range of potential glucose concentrations. Dermal interstitial fluid glucose was highly correlated with plasma glucose (r = 0.95, p < 0.0001). The mean absolute and percent differences between dermal interstitial fluid glucose and plasma glucose were 1.2 mmol/L (21 mg/dl) and 10.6%, respectively. The kinetics of dermal interstitial fluid glucose and plasma glucose were similar. There was no significant difference between dermal interstitial fluid glucose and plasma glucose in mean glucose excursion, peak glucose concentration, or time to peak glucose concentration. The correlation between dermal interstitial fluid glucose and plasma glucose was as strong as the correlation between capillary blood glucose and plasma glucose. In conclusion, dermal interstitial fluid glucose can be used to estimate plasma glucose, and has the potential to be used for monitoring patients with diabetes mellitus. PMID- 9358084 TI - Helicobacter pylori induces proinflammatory cytokines and major histocompatibility complex class II antigen in mouse gastric epithelial cells. AB - Although Helicobacter pylori has been reported to stimulate the release of various cytokines from gastric tissue, it remains unknown whether normal and nontumorous gastric epithelial cells produce these cytokines. Therefore, in this study, we used a normal mouse gastric surface mucous cell line (GSM06) to determine whether gastric epithelial cells produce proinflammatory cytokines in response to H. pylori. The expression of MHC class II antigen was also examined, to investigate whether gastric epithelial cells participate in the immune response to H. pylori. In the study, GSM06 cells were incubated with H. pylori or its lipopolysaccharide (LPS). Proinflammatory cytokines were detected by Northern and Western blot analysis. The expression of MHC class II antigen was examined by fluorescence activated cell sorter (FACS) analysis. Genetic expression of proinflammatory cytokines such as interleukin-1alpha, tumor necrosis factor alpha, and cytokine-induced neutrophil chemoattractant-2beta was enhanced by both intact and sonicated H. pylori, but not by H. pylori LPS. The expression of MHC class II antigen was induced by H. pylori more strongly than by interferon-gamma. We conclude that H. pylori induces the expression of proinflammatory cytokines and MHC class II antigen in gastric epithelial cells. Gastric epithelial cells may act as antigen-presenting cells and participate in the immune response to H. pylori infection. PMID- 9358086 TI - Transplantation of single and paired pediatric kidneys into adult recipients. AB - BACKGROUND: The transplantation of kidneys from cadaveric donors < or = 5 years of age into adult recipients is controversial. The large disparity between donor renal mass and recipient body mass is feared to be problematic. Controversy also exists whether to transplant kidneys from these young donors individually or as a pair into a single recipient. STUDY DESIGN: We retrospectively reviewed our experience from January 1991 to January 1995 with 22 adult renal transplantations using kidneys from cadaveric donors < or = 5 years of age. Ten patients received single allografts. Twelve received organs paired en bloc. Fifty-two adult recipients from cadaveric donors aged 18-55 years served as controls. All patients received cyclosporine-based immunosuppression. Recipient characteristics did not differ significantly between the groups. RESULTS: Actuarial patient and graft survival rates were similar for the two groups. The incidence of urinary complications was higher in the recipients of pediatric kidneys than in the adult donor group (18.2% versus 3.8%, respectively, p = not significant). No grafts were lost from urinary complications. Renal function, as determined by the calculated creatinine clearance, was significantly greater in the pediatric group (76.1 +/- 4.0 versus 61.4 +/- 23.2 mL/min, p = 0.035) by 6 months after transplantation. Recipients of paired pediatric kidneys initially had better renal function (63.9 +/- 21.4 mL/min) than those receiving single pediatric kidneys (38.2 +/- 11.6 mL/min) (p = 0.004), but by 6 months, no significant difference existed. At 2 years, renal function in the pediatric-donor group remained significantly better than in the adult-donor group. Hematocrit levels as a measure of erythropoiesis were similar for single pediatric, paired pediatric, and adult-donor recipients. CONCLUSIONS: Kidneys from cadaveric donors < or = 5 years of age are suitable for transplantation into adults. Pediatric kidneys provide excellent renal function despite an initially tremendous disparity between renal mass and recipient body mass. Rapid true renal growth probably occurs. No appreciable advantage is achieved by using two pediatric kidneys for a single recipient. PMID- 9358087 TI - Complications of open and laparoscopic antireflux surgery: 32-year audit at a teaching hospital. AB - BACKGROUND: Open or laparoscopic surgery for gastroesophageal reflux disease gives longterm freedom from symptoms in 83-100% of cases but has a certain percentage of complications. This study was undertaken to evaluate the early and late complication rates after primary or repeat antireflux operations. STUDY DESIGN: The records of all patients who underwent surgery for gastroesophageal reflux disease during a 32-year period at a university teaching hospital were reviewed retrospectively. Records for 793 adults (448 men and 345 women) aged 16 85 years (mean, 51) were retrieved for calculation of complication rates and statistical analysis. RESULTS: A total of 827 operations were performed: 793 primary and 41 for recurrent disease (2 patients were each reoperated on twice). There were 49 laparoscopic operations. Only two patients died (mortality, 0.3%), both after open operation. Morbidity was 24% after open surgery and 14% after laparoscopic operation. The total (early and late) complication rate was higher after reoperations than that after open or laparoscopic procedures. The overall complication rate in the open operations was similar in the first and the third decade of the study, namely, 24.6% and 26.1%, respectively. CONCLUSIONS: Surgical treatment of gastroesophageal reflux disease carries very low mortality when performed in a specialized unit. The main causes of morbidity after open operation are infectious complications. The incidence of complications is substantially lower after laparoscopic surgery than after open operation. Reoperation is seldom required, but it carries higher morbidity than the primary operations. PMID- 9358088 TI - Second surgical opinion programs: dead or alive? AB - BACKGROUND: In the 1970s, second surgical opinion programs were established in an effort to improve medical care and to control health care costs. The cost effectiveness of these programs has been questioned recently. STUDY DESIGN: A retrospective review was conducted of elective second-opinion surgical consultations for members of Local 32B-J of the International Service Employees Union for the years 1993-1994. Nonconfirmed consultations were reviewed against claims history data for the subsequent 2 years. Data were analyzed for rates of nonconfirmation by diagnosis and surgical specialty and for cost-effectiveness benefit. RESULTS: Of the 5,601 second surgical consultations performed, 490 procedures were not confirmed as medically necessary (9%). Claims history survey for these 490 patients for the 2 years following the consultation revealed that no operation was performed in 62%. The highest nonconfirmation rate (41%) was in plastic and reconstructive surgery, followed by gynecology (22%). The cost benefit ratio for the program was calculated to be 1.34. CONCLUSIONS: A second surgical opinion program confers both cognitive and psychologic beneficial effects on Joint Trust Fund members and their dependents who are advised to undergo elective operations. Our current second surgical opinion nonconfirmation rate is 9%, with hysterectomy, prostatectomy, and bunionectomy among the procedures most frequently nonconfirmed. The cost-benefit ratio was estimated at 1.34. PMID- 9358089 TI - Conservative surgery in high-risk epithelial ovarian carcinoma. AB - BACKGROUND: In epithelial ovarian cancer, conservative surgery has mainly been adopted for stage Ia disease. The aim of this study is to report on a conservative surgical approach used in selected young patients with ovarian cancer who would usually undergo radical operations. STUDY DESIGN: From 1980 through 1994, 10 patients with invasive epithelial ovarian cancer and with high grade or limited extraovarian disease were treated with conservative surgery. The mean age was 22.7 years. The stage was Ia grade 3 in 2 patients, Ic in 2 patients, IIIa in 2 patients, and IIIc in 4 patients. Eight patients were given adjuvant therapy (radiotherapy in 1 and chemotherapy in 7). RESULTS: All patients were alive and disease-free at a median followup time of 70 months (range 24-138 months). Nine patients were menstruating regularly and three had become pregnant. CONCLUSIONS: It seems that in selected patients, conservative operations can be used beyond the worldwide accepted criterion of stage Ia. This concept deserves additional investigation in larger series. PMID- 9358085 TI - Hepatic resection and transplantation for peripheral cholangiocarcinoma. AB - BACKGROUND: Recent publications have questioned the role of orthotopic liver transplantation (OLT) in treating advanced or unresectable peripheral cholangiocarcinoma (Ch-Ca). STUDY DESIGN: We reviewed our experience with Ch-Ca to determine survival rates, recurrence patterns, and risk factors in 54 patients who underwent either hepatic resection or OLT between 1981 and 1994. Liver transplantation was performed in patients with unresectable tumors (n = 12) and in those with advanced cirrhosis (n = 8). There were 33 women (61%) and 21 men (39%), with a mean age of 54.3 years. The median followup period was 6.8 years. Prognostic risk factors were analyzed by univariate and multivariate analyses. RESULTS: Mortality within 30 days was 7.4%. Overall patient and tumor-free survival rates were 64% and 57% at 1 year, 34% and 34% at 3 years, and 26% and 27% at 5 years after operation. Thirty-two patients (59.3%) experienced tumor recurrence. Univariate analysis revealed that multiple tumors, bilobar tumor distribution, regional lymph node involvement, presence of metastasis, positive surgical margins, and advanced pTNM stages were significant negative predictors of both tumor-free and patient survival. Multivariate analysis revealed that positive margins, multiple tumors, and lymph node involvement were independently associated with poor prognosis. When patients with these three negative predictors were excluded, the patient survivals at 1, 3, and 5 years were 74%, 64%, and 62%, respectively. CONCLUSIONS: Both hepatic resection and OLT are effective therapies for Ch-Ca when the tumor can be removed with adequate margins, the lesion is singular, and lymph nodes are not involved. PMID- 9358090 TI - Major vascular injuries during gynecologic laparoscopy. AB - BACKGROUND: This study was undertaken to report our experience with major vascular injuries in gynecologic laparoscopy in order to specify the circumstances under which they occurred, the means of diagnosis, the risk factors, and the means for prevention. STUDY DESIGN: Retrospective case review study. RESULTS: Seventeen patients with 21 major vascular injuries were identified. The average age of the patients was 33.8 +/- 11.6 years, and the mean body index mass was 21.6 +/- 3.08 kg/m2. Three of four of the accidents occurred during the set-up phase of laparoscopy (13 cases; 76.5%), and in 4 cases (23.5%) the accident occurred during the laparoscopic surgery procedure. Eleven (84.6%) of the complications occurring during the set-up phase were secondary to insertion of the umbilical trocar and 2 (15.4%) to insertion of the needle used to create the pneumoperitoneum (P-needle). Half (6 cases; 54.5%) of the major vascular injuries secondary to insertion of the umbilical trocar were observed when reusable trocars were used. In every case, the diagnosis was made during the operation. Two patients died, and two others presented a serious complication (phlebitis; acute ischemia requiring reoperation). CONCLUSIONS: Major vascular injuries are rare but serious complications of laparoscopic surgery. Prevention of these accidents relies on the surgeon's experience and scrupulous respect of the safety rules. In the vast majority of cases, it is necessary to convert to laparotomy immediately, calling in a vascular surgeon. PMID- 9358091 TI - Cyclical mastalgia: prevalence and impact in an outpatient breast clinic sample. AB - BACKGROUND: A descriptive study was conducted to examine prevalence of premenstrual breast symptoms, impact of cyclical mastalgia on various activities, and associated patterns of health care utilization among breast clinic outpatients. STUDY DESIGN: Patients (n = 231, age < 55 years) completed a questionnaire about lifetime and current premenstrual breast discomfort (cyclical mastalgia). RESULTS: Seventy-nine percent reported having regularly experienced cyclical breast symptoms; 48% have asked a health care provider about their mastalgia. Young women (< or = 35 years) were more than three times as likely to have had a mammogram (75%) if they regularly experienced cyclical mastalgia than if they did not (24%; p < 0.05). Current moderate to severe mastalgia lasting 5 days or more monthly was reported by 30% of women. This "clinical" level of mastalgia interferes with usual sexual activity for 33%, with physical activity for 29%, with social activity for 15%, and with work for 15% of these women. CONCLUSIONS: Reported prevalences of mastalgia obtained in this sample are higher than those reported in British studies; possible reasons for these differences are discussed. Cyclical mastalgia is a common problem sometimes severe enough to interfere with normal activity levels, and it is related to excessive use of mammography among young women. Although largely ignored both scientifically and clinically in the United States, this condition merits further biopsychosocial investigation. PMID- 9358092 TI - Primary enteric drainage of the pancreas allograft revisited. AB - BACKGROUND: Historically, primary enteric drainage (ED) of exocrine secretions in pancreas allografts was associated with a poor outcome, mostly as a result of infectious complications. On the other hand, bladder drainage (BD), which is presently used in the majority of institutions, is associated with substantial urologic morbidity. The aim of this study is to reassess the role of primary ED by reviewing our experience with ED versus BD in simultaneous pancreas-kidney transplantations. STUDY DESIGN: The records of all pancreas-kidney transplantations performed between October 1990 and September 1996 were reviewed (n = 42). Enteric drainage was used in the last 16 (38%) and BD in the first 26 (62%). The BD and ED groups were comparable with respect to donor and recipient characteristics. RESULTS: Length of stay for the transplantation (mean +/- standard deviation) was significantly shorter with ED than with BD (12.9 +/- 5.6 versus 20.4 +/- 9.6 days, p = 0.007). The total number of readmissions (1.7 +/- 1.5 versus 1.2 +/- 1.2 days, p = 0.2) and the length of hospital stay in the first 6 months after discharge (13.7 +/- 16.2 versus 10 +/- 11.3 days, p = 0.4) were similar between BD and ED. Complications requiring admission were distributed as follows in BD and ED recipients: recurrent/persistent urinary complications (46% versus 6%, p = 0.01), dehydration (27% versus 6%, p = 0.05), symptomatic graft pancreatitis (8% versus 6%, p = 0.9), gastrointestinal disturbance (27% versus 12%, p = 0.1), and wound infection (12% versus 19%, p = 0.5). The duration of the operative procedure was shorter in ED than in BD (4.3 +/- 0.9 versus 5.4 +/- 0.8 hours, p = 0.01). Reoperation during the initial transplantation stay was necessary in 23% of the patients having BD, compared with none having ED (p = 0.04). Similarly, fewer ED patients underwent reoperations compared with BD patients in the first 6 months after discharge (38% versus 69%, p = 0.04). Hospital charges for ED were lower than for BD for the initial admission ($73,458 +/- 17,103 versus $107,193 +/- 32,965, p = 0.001). Actuarial patient (96% versus 94%, p = 0.6), kidney (85% versus 87%, p = 0.9), and technically successful pancreas (90% versus 85%, p = 0.6) survival rates at 1 year were similar for BD and ED. CONCLUSIONS: Our results indicate that, compared with BD, ED is associated with less morbidity and shorter hospitalization without compromising outcome. Primary ED is a viable alternative to BD in simultaneous pancreas-kidney transplantation. More clinical experience with careful cost effectiveness analysis is needed to better assess the implications of primary ED. PMID- 9358093 TI - Prognostic study of gastric cancer without serosal invasion: reevaluation of the definition of early gastric cancer. AB - BACKGROUND: Early gastric cancer, a term defined by Japanese researchers in the 1960s, is equivalent to pT1 tumor stage regardless of nodal status. Recently, there were suggestions to exclude node-positive pT1 gastric cancer from this entity and to consider node-negative pT2 gastric cancer as early gastric cancer. STUDY DESIGN: A survival analysis was conducted of 294 patients who underwent resection for gastric cancers confined within the gastric wall (pT1, n = 164; pT2, n = 130) between 1986 and 1992. RESULTS: The cumulative 5-year survival rate was 93.5% for pT1 patients and 67.9% for pT2 patients, with an overall survival of 82.5%. There was a significant difference in the 5-year survival rate between node-positive and node-negative pT1 patients (72.8% versus 95.6%; p = 0.0095). The 5-year survival rate of node-negative pT2 patients (80.4%) was significantly worse than that of node-negative pT1 patients (p = 0.011) but was not significantly better than that of node-positive pT1 patients (p = 0.4). If excellent prognosis is a prerequisite for the definition of early gastric cancer, then node-positive pT1 cancer and node-negative pT2 cancer should not be considered early gastric cancer. CONCLUSIONS: In the 1990s, now that new imaging techniques such as endoscopic ultrasonography has been introduced, the preoperative staging of gastric cancer can be made more accurately than in the 1960s, when the term "early gastric cancer" was defined. Because the prognosis of early gastric cancers, if subcategorized by nodal status, is not homogeneously excellent, a reevaluation of the definition of early gastric cancer may be necessary. PMID- 9358094 TI - Thoracoscopic excision of mediastinal parathyroid adenomas: a report of two cases and review of the literature. AB - BACKGROUND: Most abnormal parathyroid glands can be removed through a standard cervical incision; even those in the superior mediastinum. Those located in certain areas of the mediastinum, for example posteriorly or in the aortopulmonic window, historically have required excision through a median sternotomy or thoracotomy. Angioablation is a nonsurgical alternative to management of these lesions. STUDY DESIGN: We present two case reports of mediastinal parathyroid adenomas that were excised thoracoscopically, and review the literature regarding the management of mediastinal parathyroid adenomas. RESULTS: Both patients who underwent precise localization and thoracoscopic excision of their mediastinal parathyroid adenomas had resolution of their hypercalcemia with minimal associated morbidity and shortened recovery periods. CONCLUSIONS: We suggest that thoracoscopic excision of mediastinal parathyroid adenomas is the better means of controlling hypercalcemia secondary to parathyroid adenoma in those patients considered for either median sternotomy, thoracotomy or angiographic ablation where the exact location of the lesion can be established preoperatively. PMID- 9358095 TI - Stapled pulmonary tractotomy: a rapid way to control hemorrhage in penetrating pulmonary injuries. PMID- 9358096 TI - Aortic cannulation in organ donors with pathology of the infrarenal aorta. PMID- 9358097 TI - Repair of a spigelian hernia. PMID- 9358098 TI - The "Yanni" knot: a simpler method of intracorporeal laparoscopic knot tying. PMID- 9358099 TI - Anal canal and perianal epidermoid cancers. AB - Squamous cell carcinoma of the anal canal serves as a paradigm for the successful application of multimodality treatment of solid tumors. Since 1974, multimodality treatment with combined radiation and chemotherapy has become the standard. The compelling advantage of sphincter preservation and the substantial survival benefit compared with surgery alone prompted investigators to adopt chemoradiation treatment. Several questions regarding the optimal radiation dose and chemotherapy in initial as well as salvage therapy remain, and only recently have results of several prospective randomized studies become available to address some of these unresolved issues. We reviewed the clinical aspects and historical treatment results of anal canal and perianal epidermoid cancers in light of the results of these modern trials. Current management strategies are redefined, and future directions of clinical studies are outlined. PMID- 9358100 TI - Effect of natural HIV-1 envelope V1-V2 sequence diversity on the binding of V3 specific and non-V3-specific antibodies. AB - In past years, much attention has been paid to the HIV-1 envelope (env) protein variable region 3 (V3), termed the principal neutralizing determinant. HIV-1 vaccines were often designed to target V3, and vaccine efficacy was often measured with V3-based assays. Thus, some disappointment resulted when volunteers in first clinical vaccine trials generated V3-specific antibodies that could not protect against V3-similar viruses. We describe an analysis of V1 and V2 sequence effects on antibody binding to V3 and non-V3 determinants. This study involved the preparation of seven full-length (gp160), chimeric env proteins in a vaccinia virus (VV) expression system. Chimeric proteins displayed different V1-V2 sequences but were otherwise identical. A panel of 12 monoclonal antibodies was then tested for binding activities toward the seven chimeras. Results showed that V1-V2 sequences affected antibody binding to env, both in V3 and non-V3 positions. These data demonstrate the enormous complexity of HIV-1 env protein conformation and antigenic determinants. Respect for the complexity of antibody antigen interactions encourages the design of sophisticated immunoglobulin and protein cocktails for use in HIV-1 therapies and vaccines, respectively. PMID- 9358101 TI - Rapid and selective depletion of CD4+ T lymphocytes and preferential loss of memory cells on interaction of mononuclear cells with HIV-1 glycoprotein expressing cells. AB - Contact of HIV glycoprotein-expressing cells with CD4+ T lymphocytes in vitro causes cell-cell fusion and/or cytopathogenicity. The question of whether this process similarly underlies the death of helper T cells in vivo has not yet been resolved. To investigate the loss of uninfected CD4+ T cells in an environment that may reflect the in vivo situation, unfractionated, unstimulated peripheral blood mononuclear cells were cocultured with HIV-1 glycoprotein-expressing cells, and early alterations of T-cell numbers were quantitated using a newly developed quantitative flow cytometric assay. The results demonstrate that a large fraction of normal-sized, regular CD4+ T cells disappeared immediately on cocultivation with envelope glycoprotein-expressing cells. In contrast, CD8+ T lymphocytes remained unaffected. Significant loss of uninfected T-helper cells required the presence of less than 1% infected cells. Moreover, memory T cells (CD45RO+, CD29 hi+) were depleted more rapidly than naive cells (CD45RO-, CD29 lo+). The observation that a large fraction of intact primary T-helper cells disappeared on contact with HIV glycoprotein-expressing cells suggests that a similar process may occur in vivo and contribute to the loss of T-helper cells in the infected individual. In addition, the preferential loss of memory cells may account for the early loss of immune functions in the course of HIV infection. PMID- 9358102 TI - Elevated CD38 antigen expression on CD8+ T cells is a stronger marker for the risk of chronic HIV disease progression to AIDS and death in the Multicenter AIDS Cohort Study than CD4+ cell count, soluble immune activation markers, or combinations of HLA-DR and CD38 expression. AB - The prognostic value of several immunologic markers were compared in Los Angeles Multicenter AIDS Cohort Study (MACS) participants, most of whom had been infected with HIV for >8 years. Markers studied included CD4+ cell number, flow cytometric measurements of CD8+ cell expression of CD38 and HLA-DR antigens, and serum markers of immune activation including neopterin, beta2-microglobulin, soluble interleukin-2 receptor, soluble CD8, and soluble tumor necrosis factor receptor alpha (TNF-alpha) type II. Cox proportional hazards models indicated that elevated CD38 on CD8, a flow cytometric measurement of CD8+ T-lymphocyte activation, was the most predictive marker of those studied for development of a clinical AIDS diagnosis and death. As compared with the reference group, who had CD38 on CD8 <2470 molecules per CD8+ cell and in whom 4 of 99 developed clinical AIDS within 3 years, participants with CD38 on CD8 between 2470 and 3899, 3900 and 7250, and >7250 had relative risks (and numbers developing AIDS within 3 years) of 5.0 (15 of 81), 12.3 (24 of 60), and 41.4 (36 of 49), respectively. The strong prognostic value of CD38 on CD8 measurements and the fundamental importance of chronic immune activation in the pathogenesis of HIV disease suggests that this marker might have utility in the clinical management of HIV infected persons. PMID- 9358103 TI - Prognostic value of serum HIV-RNA levels at virologic steady state after seroconversion: relation to CD4 cell count and clinical course of primary infection. AB - The objectives of this study were to evaluate the prognostic value of a single serum HIV-RNA measurement 6-24 months after HIV seroconversion, and to investigate whether any differences in outcome related to the clinical course of primary infection could be explained by presumed steady-state HIV-RNA levels at 6 24 months after seroconversion. Disease progression was analyzed by life tables and Cox proportional hazard models. A total of 93 HIV seroconverters followed for a median of 76 months (range, 19-143 months) were included in the study. The main outcome measures were development of AIDS stratified by age, the year of seroconversion, serum HIV-RNA levels, CD4 cell counts, and duration of primary illness. The proportion of patients who developed AIDS was 36% (95% confidence interval, 22%-50%) at 8 years and 50% (26%-75%) at 10 years. In the unadjusted analyses, clinical progression was significantly associated with serum HIV-RNA levels, CD4 cell counts, and duration of primary illness. The adjusted analyses indicated that HIV-RNA was the strongest predictor. Patients with long-lasting symptoms associated with primary infection had significantly higher serum HIV-RNA levels than those with less severe presentations (median counts, 11,660 vs. 2880 copies/ml, p = 0.001). It is concluded that the serum HIV-RNA level in early HIV infection is a strong independent predictor of clinical progression. Patients with long-lasting primary illnesses reach a higher viral load in steady state after seroconversion, which is probably the main reason for the poorer prognosis observed in this group of patients. PMID- 9358104 TI - Itraconazole maintenance treatment for histoplasmosis in AIDS: a prospective, multicenter trial. AB - PURPOSE: To study the efficacy and safety of maintenance treatment with itraconazole for disseminated histoplasmosis in patients with AIDS. PATIENTS AND METHODS: This was a prospective, multicenter, open-label study conducted at university-based hospitals participating in the AIDS Clinical Trial Group (ACTG). Forty-six AIDS patients with mild to moderate disseminated histoplasmosis who had successfully completed 12 weeks of induction treatment with itraconazole were treated with itraconazole, 200 mg once daily (42 patients) or 400 mg once daily (4 patients). Patients were followed at monthly intervals with clinical and laboratory assessment for relapse or toxicity. Primary outcome measures were relapse of histoplasmosis and survival. Secondary outcome measures included drug limiting toxicity and changes in serum and urine Histoplasma polysaccharide antigen (HPA) levels. RESULTS: Two patients relapsed during a median follow-up period of 87 weeks. The 1-year relapse-free rate was estimated to be 95.3% (95% CI, 85.3%-99.7%). One relapse may have been related to poor adherence to treatment and the second to concurrent administration of rifampin. From the start of maintenance treatment, the estimated 1-year survival rate was 73.0% (95% CI, 67.5%-77.9%). Five patients discontinued treatment because of suspected drug toxicity, three of whom had possible or probable hepatotoxicity. Median serum and urine HPA levels declined significantly during treatment. The only patient in whom antigen levels rose >2 U developed clinical relapse 1 week later; antigen levels were unavailable in the other relapsing patient. CONCLUSIONS: Itraconazole, 200 mg daily, is effective in preventing relapse of disseminated histoplasmosis in patients with AIDS. It is generally well tolerated, but clinicians should be alert for drug interactions and possible hepatotoxicity. PMID- 9358105 TI - Assessment of the changing willingness to participate in phase III HIV vaccine trials among men who have sex with men. AB - This paper describes the willingness of 1267 men who have sex with men (MSM) enrolled in a prospective HIV vaccine preparedness study from Chicago, Denver, and San Francisco to enroll in HIV vaccine efficacy trials. Respondents were interviewed at baseline and followed-up at 6, 12, and 18 months. At each visit respondents were tested for HIV antibodies using enzyme-linked immunosorbent assay (ELISA) testing with Western blot confirmation. Over 18 months, the annualized HIV seroincidence of this cohort was 2.4%. At baseline, 37% of the men reported that they would be "definitely" willing to participate in an HIV vaccine efficacy trial; however, this dropped to 21% at 12 months and remained stable at 18 months. Greater willingness to participate (WTP) was related to lower education, engaging in HIV risk behavior, living in Denver, white ethnicity, and older age. Changing WTP suggests that the decision to participate in HIV vaccine efficacy trials may be complex and dynamic and take an extended time. These data underscore the importance of informed consent and raise questions regarding the influence of decision-making processes on HIV vaccine efficacy trial design, compliance, and validity. PMID- 9358106 TI - Estimating the number of AIDS-defining opportunistic illness diagnoses from data collected under the 1993 AIDS surveillance definition. AB - Expansion of the surveillance definition for AIDS in the United States in 1993 caused a substantial distortion in the trend in AIDS incidence, mainly because CD4-positive (CD4+) T-lymphocyte criteria were added to the definition. To evaluate trends in the rate at which HIV-infected persons develop the opportunistic illnesses listed in the AIDS surveillance definition (AIDS-OIs), we developed a procedure for estimating the incidence of these diseases. This estimate is based primarily on the probability distributions of the time from a CD4+ count in given ranges to the diagnosis of the first AIDS-OI. Our estimates of AIDS-OI incidence change by <4% during most calendar quarters during 1991 through 1995 if we also include the estimated effects of unreported AIDS-OIs among persons with AIDS reported based on the CD4+ criteria. Our procedure eliminates the transient effect of adding the CD4+ criteria to the AIDS surveillance definition and permits us to evaluate trends in the incidence of AIDS-OIs. PMID- 9358107 TI - Redefining the growth of the heterosexual HIV/AIDS epidemic in Chicago. AB - A dramatic shift in the relative distribution of the five categories of heterosexual transmission for AIDS cases diagnosed in Chicago since 1991 prompted a mode-of-transmission validation study of what had become the most frequently reported heterosexual exposure: heterosexual relations with a person with AIDS (PWA) or documented HIV infection whose risk is not specified. METHODS: For 395 cases with originally reported heterosexual exposure, one or more of three supplemental data sources were employed: medical records were reviewed, medical providers were interviewed, and patients or proxies (i.e., spouse, significant other, or family member) were interviewed when possible. When reported HIV exposure could not be validated or reclassified, the transmission category employed was "no identifiable risk" (NIR). RESULTS: Eighty-five percent (336 of 395 cases) were reclassified into different transmission categories. Most notably, 69% (272 of 395 cases) were reclassified into transmission categories that did not involve heterosexual contact, including NIR. The cumulative percentage of cases attributable to heterosexual contact declined from 8% to 5% as a result of reclassification. Additionally, reclassification resulted in a reduction of nearly 50% in the number of AIDS cases attributable to heterosexual contact diagnosed in 1993 and 1994. CONCLUSIONS: In Chicago, an emerging problem in AIDS surveillance appears to be the use of an ambiguous heterosexual exposure category as a default when other information is not readily available. This study has found the growth in AIDS cases among persons exposed to HIV through heterosexual contact to be much slower than previously perceived. This finding may have important implications for the national debate over the extent to which heterosexual people are being infected and how funding and prevention strategies should be prioritized. PMID- 9358108 TI - Strategies for universalistic and targeted HIV prevention. AB - The controversy over "targeted" versus "universalistic" programs for HIV prevention has persisted throughout the history of the HIV/AIDS epidemic in the United States and in some European countries. Building on previous analyses, we outline methods for integrating universalistic and targeted HIV prevention programming. The outline considers possible synergy between targeted and universalistic programs, rather than a forced choice between the two. Components within this framework include a continuum of the intensity of targeted programs, specification of local risk behavior populations, categories of risk behavior, and HIV seroprevalence within local risk-behavior populations. Given the scarce resources currently available, preventing all new HIV infections is not a realistic public health goal, but with better use of current scientific knowledge, it should be possible to greatly reduce the rate of new HIV infections. PMID- 9358109 TI - The rise and fall of allergic contact dermatitis. AB - When a simple chemical hapten contacts the skin, it triggers a complex cellular and molecular reaction that eventuates in the cutaneous reaction clinically recognized as allergic contact dermatitis. This review summarizes the crucial role of the cytokines and adhesion molecules that direct the traffic of cells into, and out of, the skin after contact with an allergen. Particularly stressed are the molecular interactions between antigen-bearing Langerhans cells and responding CD4(+) T cells: this interaction results in the amplification of the memory cells that add specificity to the immune response. Finally, negative feedback mechanisms for dampening the response are briefly discussed. PMID- 9358110 TI - Electrical impedance for quantification and classification of experimental skin reactions. AB - BACKGROUND: Pathophysiologic events in biological tissue are characterized by a shift of the electrical impedance spectra of the tissue under study. OBJECTIVE: To discuss studies on experimental skin reactions with an improved impedance spectrometer. The instrumentation is related to noninvasive techniques based on other physical principles. METHODS: The results from studies on patients with allergic contact reactions (n = 8), prick tests (n = 10), and irritant contact reactions (benzalkonium chloride [n = 14], sodium lauryl sulfate [n = 12], and nonanoic acid [n = 14]), and an appropriate number of controls are reviewed. RESULTS: Results show statistically significant changes of the impedance parameters when compared with relevant controls, at different types of experimental cutaneous reactions, both allergic and irritant type. Each reaction type had a specific impedance index pattern. CONCLUSIONS: Current data indicate that the improved impedance technique offers a possible noninvasive alternative for characterization and perhaps differentiation, not only between the skin responses induced by either an allergen or an irritant, but also a capability to distinguish responses induced by chemically different irritants. The assumption that the impedance method is capable to distinguish allergic from irritant contact reactions has not been proven yet in direct comparative studies. PMID- 9358111 TI - An evaluation of the allergic contact dermatitis potential of colloidal grain suspensions. AB - BACKGROUND: Colloidal grain suspensions have been used for decades as adjuncts in the treatment of atopic dermatitis, especially in the US. In Italy, many young children have been exposed to colloidal grains. Recently, it was suggested that these bath therapies may induce allergic contact dermatitis in some young atopic children. OBJECTIVE: To evaluate the allergic skin reactions to topical oat and rice colloidal grain suspensions of normal and atopic children with and without previous exposure to colloidal grain suspensions. METHODS: A double-blind, randomized patch study. Two concentrations of oat and rice colloidal grains (0.007% and 0.7%) were applied occlusively to the backs of 65 children living in Italy, ages 6 months to 2 years (43 were atopic and 22 were normal). RESULTS: There were neither immediate urticarial nor allergic reactions in any of the 65 study subjects, atopic or nonatopic; 5 of 43 (12%) atopic subjects developed irritant reactions to the test materials. Radioallergosorbent tests (RAST) tests were performed on 55 subjects. The negative RAST test results found in the nonatopic group correlated well with nonatopic status, but positive RAST tests were found in only 8 of 35 (23%) atopic dermatitis subjects. None of the sera from positive RAST scores corresponded to subjects with irritant patch reactions. CONCLUSIONS: The data indicate that topical colloidal grains can be used as an adjunct in the management of mild atopic dermatitis in children under 2 years of age. There was no evidence of sensitization to topical colloidal grains in the group studied. PMID- 9358112 TI - Occupational skin disease in workers from the electronics industry in Singapore. AB - Of 149 workers from the electronics industry with occupational dermatoses seen at the Joint Occupational Dermatoses Clinic at the National Skin Centre, Singapore, 51.0% (76) were diagnosed to have irritant contact dermatitis, 40.9% (61) had allergic contact dermatitis, and 8.1% (12) had noncontact dermatitis. More than half of the patients were younger than 30 years of age. Common irritants were soldering flux, oils and coolants, solvents, and acids/alkalis. The most common allergens were nickel and resins, followed by rubber chemicals and the constituents of flux. Of the noncontact dermatitis, 8 were caused by occupationally relevant exacerbation of endogenous eczema, whereas there was one case each of miliaria, frictional dermatitis, and fatal toxic epidermal necrolysis caused by trichloroethylene allergy. PMID- 9358114 TI - Occupational dermatitis in the manufacture of color television tubes. AB - An outbreak of dermatitis occurred in the Flow Coat sector of a large and modern color television factory. After investigating the working procedures in this area, a risk of contamination of the skin and clothing with ammonium dichromate was found when ammonium dichromate was weighed and mixed with polyvinyl alcohol (PVA). All of the workers involved in this process were clinically examined; 9 of 18 showed dermatitis to the back of the hands and forearms. Three of these workers were transferred from the sector and were cured of their dermatitis. Patch tests were carried out using 40 allergens, including those used in the Flow Coat process. The tests for PVA and for the phosphorescent blue, green, and red pigments gave negative results at the three concentrations tested. A high incidence of sensitization was found; 9 of 18 (50%) of the workers were found to be sensitized to ammonium dichromate, which is used as a fixer. The preventive measures adopted consisted of improving the quality of the personal protective equipment (PPE) and transferring the weighing and mixing of dichromate to the company laboratory. During the next 18 months, there were no new cases of dermatitis in the Flow Coat sector. PMID- 9358115 TI - Thespesia populnea dermatitis. AB - BACKGROUND: The allergenic properties of milo wood have not been known before now. OBJECTIVE: We investigated the wood and elucidated the structure of its main sensitizer. METHODS: A bowl turner suffered from allergic contact dermatitis over years. Wood shavings were extracted, and the constituents were isolated, purified, and identified. The sensitizing capacity of the main compound was determined experimentally in guinea pigs. RESULTS: Initially, patch tests with sawdust suspensions and the isolated main constituent were positive. During the course of the study, however, the patient developed tolerance. The main component of milo wood was identified as a new mansonone: 7-hydroxy-2,3,5,6-tetrahydro-3,6, 9-trimethylnaphtho [1,8 bc] pyran-4,8-dione. In the sensitizing experiments, it showed to be a moderate sensitizer. CONCLUSION: Extensive and long-lasting exposure to certain sensitizers may cause the phenomenon of hardening as has been shown in the wood working industry. PMID- 9358116 TI - Pustular allergic contact dermatitis to isoconazole nitrate. AB - The topical imidazole antimycotics are widely used and are an infrequent cause of contact allergy. We report on a woman, who developed an unusual clinical picture of allergic contact dermatitis, namely papulo-pustular reaction, evoked by an isoconazole nitrate-containing cream. The histopathologic changes included subcorneal pustules, spongiosis, and an inflammatory infiltrate composed mainly of lymphocytes and some eosinophils and neutrophils. The patient demonstrated a severe vesicular reaction to isoconzole nitrate as is and at an imidazoles concentration of 0.5%. No cross-reactivity with other imidazole antimycotics was noted. PMID- 9358117 TI - Contact allergy and cross-reactions caused by prilocaine. AB - BACKGROUND: Allergic contact dermatitis from local amide anesthetics such as lidocaine (Xylocaine), bupivacaine (Marcaine, Bicain), mepivacaine (Carbocaine), etidocaine (Duranest), and prilocaine (Citanest) is rare. Allergic contact dermatitis from prilocaine has been reported only from EMLA cream. OBJECTIVES: We report on a patient sensitized from prilocaine used as a local infiltration anesthetic. METHODS: Conventional patch testing with various local amide anesthetics as well as intracutaneous tests with amide anesthetics were performed. RESULTS: Patch testing with EMLA cream (as is) (Astra, Sweden), containing prilocaine and lidocaine provoked an allergic patch test reaction, whereas patch testing with lidocaine was negative. Patch testing with Citanest containing 2% prilocaine and adrenaline was positive. Also articaine gave an allergic patch test reaction, whereas mepivacaine, benzocaine, and caine-mix were negative. Intracutaneous tests with prilocaine and articaine were positive. CONCLUSION: Our patient had been sensitized from prilocaine and cross-reacted to articaine, but not to other amide anesthetics. Cross-reactions between prilocaine and amide anesthetics were reviewed. As there seem to be no clear rules regarding cross-reactivity among various amide anesthetics, it is safest to test as many amide type anesthetics as possible in case of a delayed type allergy to any member of this family. PMID- 9358118 TI - Allergic contact dermatitis from hydrocolloid dressings. AB - BACKGROUND: Hydrocolloid wound dressings have been in use for nearly two decades, and have rarely caused allergic contact dermatitis. DuoDERM E (DuoDERM CGF) is a newer version of DuoDERM (ConvaTec Ltd, a division of Bristol-Myers Squibb Co, Princeton, NJ) that contains a sensitizing derivative of colophony. OBJECTIVE: We describe three patients who developed eczematous lesions under this type of wound covering. METHODS: The patients were patch tested to the European standard series, to a glues and adhesives series, and to pieces of various adhesive dressings. RESULTS: The patients displayed positive patch tests to colophony and to DuoDERM E or DuoDERM CGF hydrocolloid dressings. CONCLUSION: These dressings contain the pentaerythritol ester of hydrogenated rosin as a tackifying agent, and this substance retains the sensitizing potential of colophony. The addition of this compound is an important change that may negatively alter the good safety record of ConvaTec dressings. PMID- 9358119 TI - Is it really fragrance-free? AB - BACKGROUND: Fragrance allergy is the most common cause of cosmetic contact dermatitis. Many occult sources of fragrance exist. Those which cause the most concern are some "fragrance-free" products that contain fragrance raw ingredients. Thus, the very patients requiring fragrance-free items may be exposed to potential perfume allergens or cross-reactors in seemingly safe products. Additionally, medications dermatologists recommend, both prescription and over-the-counter, sometimes contain fragrance. OBJECTIVE: This report describes a patient with chronic hand dermatitis sensitized to multiple fragrance ingredients including rose oil, present in the "fragrance-free" soap she used. Additionally, the tar soaks recommended to her also contained fragrance. CONCLUSION: It is no longer sufficient to recommend the use of products labeled fragrance-free to fragrance-sensitive patients. These patients must be educated to read labels and look for plant extracts that are potential perfume sensitizers and cross-reactors. Rose oil, which has been felt to be a rare sensitizer, may be a more common allergen than previously recognized, perhaps because of its existence in a popular "fragrance-free" soap and, conceivably, in many "all natural" products. Further testing with rose oil should be conducted in the future. Finally, manufacturers need to be more forthright in the labeling of their products. PMID- 9358120 TI - Lawyers, regulations, and cosmetic claims. PMID- 9358121 TI - Sunscreen update. PMID- 9358122 TI - Allergic contact dermatitis from aromatherapy. AB - Allergic contact dermatitis secondary to aromatherapy has been only rarely reported. We present 39-year-old woman who had used aromatherapy products for approximately 2 to 3 years who presented with an erythematous eruption on her face and chest. Patch testing showed a positive reaction to neomycin and fragrance mix. On cessation of her aromatherapy products, her eruption rapidly resolved. Aromatherapy products containing essential oils may need to be considered as a cause of allergic contact dermatitis because of the increasing popularity of this treatment. PMID- 9358126 TI - A comparison of recombinant hirudin with a low-molecular-weight heparin to prevent thromboembolic complications after total hip replacement. AB - BACKGROUND: Patients who undergo total hip replacement have a high risk of thromboembolic complications. Recombinant hirudin (desirudin), a specific inhibitor of thrombin, represents a new development in antithrombotic therapy. We compared the efficacy and safety of desirudin with those of a low-molecular weight heparin (enoxaparin) for the prevention of thromboembolic complications in patients undergoing primary total hip replacement. METHODS: Both treatments, which were assigned in a randomized, double-blind manner, were started preoperatively: enoxaparin on the evening before surgery, and desirudin within 30 minutes before the start of surgery. The dose of desirudin was 15 mg subcutaneously twice daily, and the dose of enoxaparin was 40 mg subcutaneously once daily. The duration of treatment was 8 to 12 days. Deep-vein thrombosis was verified by bilateral venography performed at the end of the treatment period or earlier, if there were clinical signs of deep-vein thrombosis. RESULTS: At 31 centers in 10 European countries, 2079 eligible patients were randomly assigned to receive desirudin or enoxaparin. A total of 1587 patients were included in the primary analysis of efficacy. In the desirudin group, as compared with the enoxaparin group, there was a significantly lower rate of proximal deep-vein thrombosis (4.5 vs. 7.5 percent, P=0.01; relative reduction in risk, 40.3 percent) and a lower overall rate of deep-vein thrombosis (18.4 vs. 25.5 percent, P=0.001; relative reduction in risk, 28.0 percent). The safety profiles were similar in the two treatment groups. CONCLUSIONS: When administered 30 minutes before total hip replacement surgery, desirudin is more effective than enoxaparin in preventing deep-vein thrombosis. PMID- 9358127 TI - Color duplex ultrasonography in the diagnosis of temporal arteritis. AB - BACKGROUND: The diagnosis of temporal arteritis usually requires a biopsy of the temporal artery. We examined the usefulness of color duplex ultrasonography in patients suspected of having temporal arteritis. METHODS: In this prospective study, all patients seen in the departments of rheumatology and ophthalmology from January 1994 to October 1996 who had clinically suspected active temporal arteritis or polymyalgia rheumatica were examined by duplex ultrasonography. The final diagnoses, made according to standard criteria, were temporal arteritis in 30 patients, 21 with biopsy-confirmed disease; polymyalgia rheumatica in 37; and negative histologic findings and a diagnosis other than temporal arteritis or polymyalgia rheumatica in 15. We also studied 30 control patients matched for age and sex to the patients with arteritis. Two ultrasound studies were performed and read before the biopsies; one ultrasonographer was unaware of the clinical information. RESULTS: In 22 (73 percent) of the 30 patients with temporal arteritis, ultrasonography showed a dark halo around the lumen of the temporal arteries. The halos disappeared after a mean of 16 days (range, 7 to 56) of treatment with corticosteroids. Twenty-four patients (80 percent) had stenoses or occlusions of temporal-artery segments, and 28 patients (93 percent) had stenoses, occlusions, or a halo. No halos were identified in the 82 patients without temporal arteritis; 6 (7 percent) had stenoses or occlusions. For each of the three types of abnormalities identified by ultrasonography, the interrater agreement was > or =95 percent. CONCLUSIONS: There are characteristic signs of temporal arteritis that can be visualized by color duplex ultrasonography. The most specific sign is a dark halo, which may be due to edema of the artery wall. In patients with typical clinical signs and a halo on ultrasonography, it may be possible to make a diagnosis of temporal arteritis and begin treatment without performing a temporal-artery biopsy. PMID- 9358129 TI - Sexually transmitted infection as a cause of anal cancer. AB - BACKGROUND: The incidence of anal cancer has increased in recent decades, particularly among women. To identify underlying risk factors, we conducted a population-based case-control study in Denmark and Sweden. METHODS: We conducted telephone interviews with 324 women and 93 men in whom invasive or in situ anal cancer was diagnosed between 1991 and 1994, 534 controls with adenocarcinoma of the rectum, and 554 population controls. The interviews covered a wide spectrum of possible risk factors for anal cancer. Odds ratios were calculated by logistic regression. Specimens of anal-cancer tissue and samples of rectal adenocarcinomas were tested for human papillomavirus (HPV) DNA with the polymerase chain reaction. RESULTS: Multivariate analysis revealed consistent and statistically significant associations between measures of sexual promiscuity and the risk of anal cancer in both men and women. There was a significant trend toward an association between higher numbers of partners of the opposite sex in women (P<0.001) and men (P<0.05) and strong associations with a variety of venereal diseases. In women, receptive anal intercourse, particularly before the age of 30 years, and venereal infections in the partner were also associated with an increased risk (odds ratios, 3.4 and 2.4, respectively). Fifteen percent of the men with anal cancer reported having had homosexual contact, as compared with none of the controls (P<0.001). High-risk types of HPV, notably HPV-16, were detected in 84 percent of the anal-cancer specimens examined, whereas all rectal adenocarcinoma specimens tested were negative for HPV. CONCLUSIONS: Our study provides strong evidence that a sexually transmitted infection causes anal cancer. The presence of high-risk types of HPV, notably HPV-16 (which is known to cause cancer of the cervix), in the majority of anal-cancer tissue specimens suggests that most anal cancers are potentially preventable. PMID- 9358128 TI - Human papillomavirus infection in women infected with the human immunodeficiency virus. AB - BACKGROUND: Among women infected with the human immunodeficiency virus (HIV), there is a high prevalence of human papillomavirus (HPV) infections. However, little is known about the natural history of HPV infections in HIV-seropositive women, and persistent HPV infections may explain the increased risk of cervical squamous intraepithelial lesions and invasive cervical cancer in HIV-seropositive women. METHODS: A total of 220 HIV-seropositive and 231 HIV-seronegative women in the New York City area were evaluated at two or more semiannual gynecologic examinations that included a Pap test, a test for HPV DNA, and colposcopy. RESULTS: HPV DNA was detected at the initial examination in 56 percent of the HIV seropositive and 31 percent of the HIV-seronegative women. After four examinations, the cumulative prevalence of HPV infection was 83 percent in the seropositive women and 62 percent in the seronegative women (P<0.001). Persistent HPV infections were found in 24 percent of the seropositive women but in only 4 percent of the seronegative women (P<0.001). Twenty percent of the seropositive women and 3 percent of the seronegative women had persistent infections with HPV 16-associated viral types (16, 31, 33, 35, or 58) or HPV-18-associated types (18 or 45) (P<0.001), which are most strongly associated with cervical cancer. The detection of HPV DNA in women with previously negative tests was not associated with sexual activity during the interval since the preceding examination. CONCLUSIONS: HIV-seropositive women have a high rate of persistent HPV infections with the types of HPV that are strongly associated with the development of high grade squamous intraepithelial lesions and invasive cervical cancer. These persistent infections may explain the increased incidence of squamous intraepithelial lesions in HIV-seropositive women. PMID- 9358130 TI - Images in clinical medicine. Pancoast's syndrome. PMID- 9358132 TI - Superior pulmonary sulcus tumors and Pancoast's syndrome. PMID- 9358131 TI - Shattuck lecture--cardiovascular medicine at the turn of the millennium: triumphs, concerns, and opportunities. PMID- 9358133 TI - Clinical problem-solving. High on the differential. PMID- 9358134 TI - Antithrombotic agents and thromboembolic disease. PMID- 9358135 TI - Sonography in giant-cell arteritis. PMID- 9358136 TI - Human papillomaviruses and anogenital cancers. PMID- 9358137 TI - Effect of inhaled formoterol and budesonide on exacerbations of asthma. Formoterol and Corticosteroids Establishing Therapy (FACET) International Study Group. AB - BACKGROUND: The role of long-acting, inhaled beta2-agonists in treating asthma is uncertain. In a double-blind study, we evaluated the effects of adding inhaled formoterol to both lower and higher doses of the inhaled glucocorticoid budesonide. METHODS: After a four-week run-in period of treatment with budesonide (800 microg twice daily), 852 patients being treated with glucocorticoids were randomly assigned to one of four treatments given twice daily by means of a dry powder inhaler (Turbuhaler): 100 microg of budesonide plus placebo, 100 microg of budesonide plus 12 microg of formoterol, 400 microg of budesonide plus placebo, or 400 microg of budesonide plus 12 microg of formoterol. Terbutaline was permitted as needed. Treatment continued for one year; we compared the frequency of exacerbations of asthma, symptoms, and lung function in the four groups. A severe exacerbation was defined by the need for oral glucocorticoids or a decrease in the peak flow to more than 30 percent below the base-line value on two consecutive days. RESULTS: The rates of severe and mild exacerbations were reduced by 26 percent and 40 percent, respectively, when formoterol was added to the lower dose of budesonide. The higher dose of budesonide alone reduced the rates of severe and mild exacerbations by 49 percent and 37 percent, respectively. Patients treated with formoterol and the higher dose of budesonide had the greatest reductions -- 63 percent and 62 percent, respectively. Symptoms of asthma and lung function improved with both formoterol and the higher dose of budesonide, but the improvements with formoterol were greater. CONCLUSIONS: In patients who have persistent symptoms of asthma despite treatment with inhaled glucocorticoids, the addition of formoterol to budesonide therapy or the use of a higher dose of budesonide may be beneficial. The addition of formoterol to budesonide therapy improves symptoms and lung function without lessening the control of asthma. PMID- 9358138 TI - A comparison of low-dose inhaled budesonide plus theophylline and high-dose inhaled budesonide for moderate asthma. AB - BACKGROUND: Inhaled glucosteroids and oral theophylline are widely used to treat asthma. We compared the benefits of adding theophylline to inhaled glucosteroid with those of doubling the dose of inhaled glucosteroid in patients with persistent symptoms despite the use of inhaled glucosteroid. METHODS: In a double blind, placebo-controlled trial, we randomly assigned 62 patients to receive either 400 microg of inhaled budesonide (low-dose budesonide) with 250 or 375 mg of theophylline (depending on body weight) or 800 microg of inhaled budesonide (high-dose budesonide). All doses were given twice daily for three months. Lung function was measured serially, and patients kept records of peak expiratory flow, symptoms, and albuterol use. The effects of treatment on endogenous cortisol levels were also assessed. RESULTS: Both treatments resulted in improvements in lung function that were sustained throughout the study. As compared with treatment with high-dose budesonide, treatment with low-dose budesonide plus theophylline resulted in greater improvements in forced vital capacity (P=0.03) and forced expiratory volume in one second (P= 0.03). There were significant and similar reductions in beta2-agonist use and the variability of peak expiratory flow, a correlate of bronchial hyperresponsiveness and the severity of asthma. Serum cortisol concentrations were significantly reduced in the group given high-dose budesonide (from a mean [+/-SE] of 18.4+/-2.4 microg per deciliter to 15.9+/-2.1 microg per deciliter, P=0.02) but were unchanged in the other group. The median serum theophylline concentration was 8.7 microg per milliliter (therapeutic range, 10 to 20) among those who received theophylline. Both treatments were well tolerated. CONCLUSIONS: For patients with moderate asthma and persistent symptoms, low-dose inhaled budesonide with theophylline and high-dose inhaled budesonide produced similar benefits. Effects were achieved at theophylline concentrations below the recommended therapeutic range. The addition of low-dose theophylline to inhaled glucosteroid may be preferable to and cheaper than increasing the dose of inhaled glucosteroid. PMID- 9358140 TI - Propranolol therapy for ectopic beta-adrenergic receptors in adrenal Cushing's syndrome. PMID- 9358139 TI - Pathophysiology of premature skin aging induced by ultraviolet light. AB - BACKGROUND: Long-term exposure to ultraviolet irradiation from sunlight causes premature skin aging (photoaging), characterized in part by wrinkles, altered pigmentation, and loss of skin tone. Photoaged skin displays prominent alterations in the collagenous extracellular matrix of connective tissue. We investigated the role of matrix-degrading metalloproteinases, a family of proteolytic enzymes, as mediators of collagen damage in photoaging. METHODS: We studied 59 whites (33 men and 26 women, ranging in age from 21 to 58 years) with light-to-moderate skin pigmentation, none of whom had current or prior skin disease. Only some of the participants were included in each of the studies. We irradiated their buttock skin with fluorescent ultraviolet lights under standard conditions and obtained skin samples from irradiated and nonirradiated areas by keratome or punch biopsy. In some studies, tretinoin and its vehicle were applied to skin under occlusion 48 hours before ultraviolet irradiation. The expression of matrix metalloproteinases was determined by in situ hybridization, immunohistology, and in situ zymography. Irradiation-induced degradation of skin collagen was measured by radioimmunoassay of soluble cross-linked telopeptides. The protein level of tissue inhibitor of matrix metalloproteinases type 1 was determined by Western blot analysis. RESULTS: A single exposure to ultraviolet irradiation increased the expression of three matrix metalloproteinases -- collagenase, a 92-kd gelatinase, and stromelysin -- in skin connective tissue and outer skin layers, as compared with nonirradiated skin. The degradation of endogenous type I collagen fibrils was increased by 58 percent in irradiated skin, as compared with nonirradiated skin. Collagenase and gelatinase activity remained maximally elevated (4.4 and 2.3 times, respectively) for seven days with four exposures to ultraviolet irradiation, delivered at two-day intervals, as compared with base-line levels. Pretreatment of skin with tretinoin (all-trans retinoic acid) inhibited the induction of matrix metalloproteinase proteins and activity (by 70 to 80 percent) in both connective tissue and outer layers of irradiated skin. Ultraviolet irradiation also induced tissue inhibitor of matrix metalloproteinases-1, which regulates the enzyme. Induction of the inhibitor was not affected by tretinoin. CONCLUSIONS: Multiple exposures to ultraviolet irradiation lead to sustained elevations of matrix metalloproteinases that degrade skin collagen and may contribute to photoaging. Treatment with topical tretinoin inhibits irradiation-induced matrix metalloproteinases but not their endogenous inhibitor. PMID- 9358141 TI - Images in clinical medicine. Cutaneous melanoma metastases. PMID- 9358143 TI - Pernicious anemia. PMID- 9358142 TI - Fifty years later: the significance of the Nuremberg Code. PMID- 9358145 TI - Controversial Journal editorials. PMID- 9358144 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 35-1997. A 65-year-old woman with a dry cough and pulmonary nodules. PMID- 9358146 TI - Asthma disease management. PMID- 9358147 TI - Understanding premature skin aging. PMID- 9358149 TI - The ups and downs of nucleic acid duplex stability: structure-stability studies on chemically-modified DNA:RNA duplexes. AB - In an effort to discover novel oligonucleotide modifications for antisense therapeutics, we have prepared oligodeoxyribonucleotides containing more than 200 different modifications and measured their affinities for complementary RNA. These include modifications to the heterocyclic bases, the deoxy-ribose sugar and the phosphodiester linkage. From these results, we have been able to determine structure-activity relationships that correlate hybridization affinity with changes in oligonucleotide structure. Data for oligonucleotides containing modified pyrimidine nucleotides are presented. In general, modifications that resulted in the most stable duplexes contained a heteroatom at the 2'-position of the sugar. Other sugar modifications usually led to diminished hybrid stability. Most backbone modifications that led to improved hybridization restricted backbone mobility and resulted in an A-type sugar pucker for the residue 5'to the modified internucleotide linkage. Among the heterocycles, C-5-substituted pyrimidines stood out as substantially increasing duplex stability. PMID- 9358150 TI - BplI, a new BcgI-like restriction endonuclease, which recognizes a symmetric sequence. AB - Bcg I and Bcg I-like restriction endonucleases cleave double stranded DNA specifically on both sides of their asymmetric recognition sequences which are interrupted by several ambiguous base pairs. Their heterosubunit structure, bifunctionality and stimulation by AdoMet make them different from other classified restriction enzymes. Here we report on a new Bcg I-like restriction endonuclease, Bpl I from Bacillus pumilus , which in contrast to all other Bcg I like enzymes, recognizes a symmetric interrupted sequence, and which, like Bcg I, cleaves double stranded DNA upstream and downstream of its recognition sequence (8/13)GAGN5CTC(13/8). Like Bcg I, Bpl I is a bifunctional enzyme revealing both DNA cleavage and methyltransferase activities. There are two polypeptides in the homogeneous preparation of Bpl I with molecular masses of approximately 74 and 37 kDa. The sizes of the Bpl I subunits are close to those of Bcg I, but the proportion 1:1 in the final preparation is different from that of 2:1 in Bcg I. Low activity observed with Mg2+increases >100-fold in the presence of AdoMet. Even with AdoMet though, specific cleavage is incomplete. S -adenosylhomocysteine (AdoHcy) or sinefungin can replace AdoMet in the cleavage reaction. AdoHcy activated Bpl I yields complete cleavage of DNA. PMID- 9358151 TI - Oligonucleotide dendrimers: synthesis and use as polylabelled DNA probes. AB - Oligonucleotide dendrimers were synthesized using a novel phosphoramidite synthon, tris-2,2,2-[3-(4,4'-dimethoxytrityloxy) propyloxymethyl]ethyl- N , N diisopropylaminocyanethoxy phosphoramidite. Label, incorporated using [gamma 32P]ATP and polynucleotide kinase, was increased in proportion to the number of 5'-ends. There was a similar increase in signal when these multiply labelled oligonucleotides were used as probes to oligonucleotide arrays. A dendrimeric oligonucleotide was used successfully as a primer in the PCR. The strand bearing the dendrimer was resistant to degradation by T7 Gene 6 exonuclease making it easy to convert the double-stranded product of the PCR to a multiply-labelled, single-stranded probe. PMID- 9358152 TI - The ETS family member ERM contains an alpha-helical acidic activation domain that contacts TAFII60. AB - Transcription factors are modular entities built up of discrete domains, some devoted to DNA binding and others permitting transcriptional modulation. The structure of DNA binding domains has been thoroughly investigated and structural classes clearly defined. In sharp contrast, the structural constraints put on transactivating regions, if any, are mostly unknown. Our investigations focus on ERM, a eukaryotic transcription factor of the ETS family. We have previously shown that ERM harbours two transactivating domains (TADs) with distinct functional features: AD1 lies in the first 72 amino acids of ERM, while AD2 sits in the last 62. Here we show that AD1 is a bona fide acidic TAD, for it activated transcription in yeast cells, while AD2 did not. AD1 contains a 20 amino acid stretch predicted to form an alpha-helix that is found unchanged in the related PEA3 and ER81 transcription factors. Circular dichroism analysis revealed that a 32 amino acid peptide encompassing this region is unstructured in water but folds into a helix when the hydrophobic solvent trifluoroethanol is added. The isolated helix was sufficient to activate transcription and mutations predicted to disrupt it dramatically affected AD1-driven transactivation, whereas mutations decreasing its acidity had more gentle effects. A phenylalanine residue within the helix was particularly sensitive to mutations. Finally, we observed that ERM bound TAFII60 via AD1 and bound TBP and TAFII40, presumably via other activation domains. PMID- 9358153 TI - Functional recognition of fragmented operator sites by R17/MS2 coat protein, a translational repressor. AB - The R17/MS2 coat protein serves as a translational repressor of replicase by binding to a 19 nt RNA hairpin containing the Shine-Dalgarno sequence and the initiation codon of the replicase gene. We have explored the structural features of the RNA operator site that are necessary for efficient translational repression by the R17/MS2 coat protein in vivo . The R17/MS2 coat protein efficiently directs lysogen formation for P22 R17 , a bacteriophage P22 derivative that carries the R17/MS2 RNA operator site within the P22 phage ant mRNA. Phages were constructed that contain fragmented operator sites such that the Shine-Dalgarno sequence and the initiation codon of the affected gene are not located within the RNA hairpin. The wild-type coat protein directs efficient lysogen formation for P22 phages that carry several fragmented RNA operator sites, including one in which the Shine-Dalgarno sequence is positioned 4 nt outside the coat protein binding site. Neither the wild-type R17/MS2 coat protein nor super-repressor mutants induce lysogen formation for a P22 phage encoding an RNA hairpin at a distance of 9 nt from the Shine-Dalgarno sequence, implying that a discrete region of biological repression is defined by the coat protein-RNA hairpin interaction. The assembly of RNA species into capsid structures is not an efficient means whereby the coat protein achieves translational repression of target mRNA transcripts. The R17/MS2 coat protein exerts translational regulation that extends considerably beyond the natural biological RNA operator site structure; however, the coat protein still mediates repression in these constructs by preventing ribosome access to linear sequence determinants of the translational initiation region by the formation of a stable RNA secondary structure. An efficient translational regulatory mechanism in bacteria appears to reside in the ability of proteins to regulate RNA folding states for host cell and phage mRNAs. PMID- 9358156 TI - Binding of Hoechst 33258 to the TAR RNA of HIV-1. Recognition of a pyrimidine bulge-dependent structure. AB - The transactivation response region (TAR) RNA is an essential component in transcriptional regulation of the human immunodeficiency virus type-1 (HIV-1) genome. We have examined the interaction between TAR RNA and the bisbenzimidazole derivative Hoechst 33258. Previous studies have shown that this drug, which is well known as an AT-selective DNA minor groove binder, can also interact with GC rich sequences in DNA as well as with RNA, possibly by intercalation. Absorption spectroscopy, circular dichroism and electric linear dichroism, as well as RNase A footprinting, were employed to compare binding of Hoechst 33258 to wild-type RNA and its analogue lacking the pyrimidine bulge. The uridine bulge, which is an important contributor to the structural stability of TAR, plays an essential role in drug binding. Deletion of the bulge destabilizes both free and drug-bound forms of TAR and markedly affects the orientation of the drug chromophore complexed with the RNA. According to the linear dichroism data, the bisbenzimidazole is oriented more or less perpendicular to the RNA helix axis. The data are compatible with a model in which the bisbenzimidazole chromophore is inserted into the existing cavity created by the pyrimidine bulge. The footprinting experiments, showing that the drug binds to a unique site opposite the unpaired uridine residues, also support this model. The binding of Hoechst 33258 to the sequence 5'-GCUCU, which delimits the cavity, reflects the greater accessibility of that region compared with other sites in the RNA molecule. The identification of a binding site for small molecules in TAR offers promising perspectives for developing drugs that would block the access of TAR RNA to proteins and therefore for the design of anti-HIV agents. PMID- 9358155 TI - TAR RNA decoys inhibit tat-activated HIV-1 transcription after preinitiation complex formation. AB - The ability of the HIV-1 Tat protein to trans -activate HIV-1 transcription in vitro is specifically inhibited by a circular TAR RNA decoy. This inhibition is not overcome by adding an excess of Tat to the reaction but is partially overcome by adding Tat in combination with nuclear extract, suggesting that TAR RNA might function by interacting with a complex containing Tat and cellular factor(s). A cell-free transcription system involving immobilized DNA templates was used to further define the factor(s) that interact with TAR RNA. Preinitiation complexes formed in the presence or absence of Tat were purified on immobilized templates containing the HIV-1 promoter. After washing, nucleotides and radiolabelled UTP were added and transcription was measured. The presence of Tat during preinitiation complex formation resulted in an increase in the level of full length HIV-1 transcripts. This Tat-activated increase in HIV-1 transcription was not inhibited by circular TAR decoys added during preinitiation complex formation but was inhibited by circular TAR decoys subsequently added during the transcription reaction. These results suggest that TAR decoys inhibit Tat activated HIV-1 transcription after preinitiation complex formation, perhaps by interacting with components of transcription complexes. PMID- 9358154 TI - The motif V of plum pox potyvirus CI RNA helicase is involved in NTP hydrolysis and is essential for virus RNA replication. AB - The plum pox potyvirus (PPV) protein CI is an RNA helicase whose function in the viral life cycle is still unknown. The CI protein contains seven conserved sequence motifs typical of RNA helicases of the superfamily SF2. We have introduced several individual point mutations into the region coding for motif V of the PPV CI protein and expressed these proteins in Escherichia coli as maltose binding protein fusions. Mutations that abolished RNA helicase activity also disturbed NTP hydrolysis. No mutations affected the RNA binding capacity of the CI protein. These mutations were also introduced in the PPV genome making use of a full-length cDNA clone. Mutant viruses carrying CI proteins with reduced RNA helicase activity replicated very poorly in protoplasts and were unable to infect whole plants without rapid pseudoreversion to wild-type. These results indicate that motif V is involved in the NTP hydrolysis step required for potyvirus RNA helicase activity, and that this activity plays an essential role in virus RNA replication inside the infected cell. PMID- 9358158 TI - New dye-labeled terminators for improved DNA sequencing patterns. AB - We have used two new dye sets for automated dye-labeled terminator DNA sequencing. One set consists of four, 4,7-dichlororhodamine dyes (d-rhodamines). The second set consists of energy-transfer dyes that use the 5-carboxy-d rhodamine dyes as acceptor dyes and the 5- or 6-carboxy isomers of 4' aminomethylfluorescein as the donor dye. Both dye sets utilize a new linker between the dye and the nucleotide, and both provide more even peak heights in terminator sequencing than the dye-terminators consisting of unsubstituted rhodamine dyes. The unsubstituted rhodamine terminators produced electropherograms in which weak G peaks are observed after A peaks and occasionally C peaks. The number of weak G peaks has been reduced or eliminated with the new dye terminators. The general improvement in peak evenness improves accuracy for the automated base-calling software. The improved signal-to-noise ratio of the energy-transfer dye-labeled terminators combined with more even peak heights results in successful sequencing of high molecular weight DNA templates such as bacterial artificial chromosome DNA. PMID- 9358157 TI - The yeast gene YNL292w encodes a pseudouridine synthase (Pus4) catalyzing the formation of psi55 in both mitochondrial and cytoplasmic tRNAs. AB - The protein products of two yeast Saccharomyces cerevisiae genes (YNL292w and CBF5) display a remarkable sequence homology with Escherichia coli tRNA:pseudouridine-55 synthase (encoded by gene truB). The gene YNL292w coding for one of these proteins was cloned in an E.coli expression vector downstream of a His6-tag. The resulting recombinant protein (Pus4) was expressed at high level and purified to homogeneity by metal affinity chromatography on Ni2+-NTA-agarose, followed by ion-exchange chromatography on MonoQ. The purified Pus4p catalyzes the formation of pseudouridine-55 in T7 in vitro transcripts of several yeast tRNA genes. In contrast to the known yeast pseudouridine synthase (Pus1) of broad specificity, no other uridines in tRNA molecules are modified by the cloned recombinant tRNA:Psi55 synthase. The disruption of the corresponding gene YNL292w in yeast, which has no significant effect on the growth of yeast cells, leads to the complete disappearance of the Psi55 formation activity in a cell-free extract. These results allow the formal identification of the protein encoded by the yeast ORF YNL292w as the only enzyme responsible for the formation of Psi55 which is almost universally conserved in tRNAs. The substrate specificity of the purified YNL292w-encoded recombinant protein was shown to be similar to that of the native protein present in yeast cell extract. Chemical mapping of pseudouridine residues in both cytoplasmic and mitochondrial tRNAs from the yeast strain carrying the disrupted gene reveals that the same gene product is responsible for Psi55 formation in tRNAs of both cellular compartments. PMID- 9358159 TI - Replication initiation and elongation fork rates within a differentially expressed human multicopy locus in early S phase. AB - Replication of the 400 copies of the 43 kb human ribosomal RNA (rDNA) locus spans most of the S phase. To examine the basis for the unusual pattern of rDNA replication, a sensitive strategy was developed to map origins of DNA replication and measure apparent rates of fork progression within a chromosomal locus. This technique, termed differential intragenomic replication timing, revealed that initiation within the actively transcribed rDNA occurred in early S within a 10.7 kb region spanning the promoter and 5' external transcribed spacer. Forks emanating from this early bidirectional origin progressed at an apparent slow rate with the sense and anti-sense forks moving at 0.32 and 0.23 kb/min. Using a photochemical-based technique, the chromatin status of the rDNA repeats was assayed throughout the S phase. Approximately 85% of the rDNA repeats were in a transcriptionally active chromatin structure at the start of S phase. A progressive decrease in the transcription state of the rDNA loci was observed, reaching a minimum between 3 and 6 h in mid S phase. Altogether, the data suggest a link between RNA polymerase I mediated transcription and site-specific initiation of DNA replication within the rDNA multicopy locus. PMID- 9358160 TI - Characterization of multiple alternative RNAs resulting from antisense transcription of the PR264/SC35 splicing factor gene. AB - The PR264/SC35 splicing factor belongs to the family of SR proteins which function as essential and alternative splicing factors. Here, we report that the human PR264/SC35 locus is bidirectionally transcribed. Double in situ hybridization experiments have allowed simultaneous detection of sense and antisense RNA in human CCRF-CEM cells, suggesting that expression of the corresponding genes is not mutually exclusive. We have characterized three main classes of ET RNAs encoded by the opposite strand of the PR264/SC35 gene and containing PR264/SC35-overlapping sequences, PR264/SC35-non overlapping sequences or a combination of both. We show that their expression results from the use of alternative promoters, exons and polyadenylation signals. PR264/SC35-non overlapping ET mRNA species potentially encode two protein isoforms (449 and 397 amino acids) and are expressed from the PR264/SC35 promoting region. Northern blots and RNase protection analyses indicate that ET polyadenylated RNAs are differentially expressed in several human cell lines. Similar studies performed in the mouse have revealed that the bidirectional transcription of the PR264/SC35 locus is a conserved mechanism and that the open reading frame identified in a subset of human ET mRNAs is highly conserved (93% homology). Northern blot analyses performed with several murine tissues confirmed the differential expression of the ET gene and revealed that it is predominantly expressed in the testis. PMID- 9358161 TI - NMR determination of the conformational and drug binding properties of the DNA heptamer d(GpCpGpApApGpC) in aqueous solution. AB - 1D and 2D NMR spectroscopy (500/600 MHz) has been used to investigate the equilibrium conformational states of the deoxyheptanucleotide 5' d(GpCpGpApApGpC), as well as its complexation with the phenanthridinium drug ethidium bromide (EB). Quantitative determination (reaction constants and thermodynamic parameters) of the conformational equilibrium of the heptamer in solution and its complexation with EB was based on analysis of the dependence of proton chemical shifts on concentration (at two temperatures, 298 and 308 K) and on temperature (in the range 278-353 K). The experimental results were analysed in terms of a model of the dynamic equilibrium between single-stranded, hairpin and bulged dimer forms of the deoxyheptanucleotide and its complexes with EB. Calculation of the relative amounts of the different complexes reveals important features of the dynamic equilibrium as a function of both temperature and the ratio of the drug and heptamer concentrations. The quantitative analysis also provides the limiting proton chemical shifts of EB in each complex which have been used to determine the most favourable structures of the intercalated complexes of EB with the (GC) sites of both the hairpin and dimer forms of the heptanucleotide. PMID- 9358162 TI - Higher fidelity of RNA-dependent DNA mispair extension by M184V drug-resistant than wild-type reverse transcriptase of human immunodeficiency virus type 1. AB - Reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) has low fidelity compared with RTs of other retroviruses and cellular DNA polymerases. We and others have previously found that the fidelity of DNA-dependent DNA polymerization (DDDP) of M184V-mutated HIV-1 RT is significantly higher than that of wild-type RT. Viruses containing the M184V substitution are highly resistant to (-)-2'-dideoxy-3'-thiacytidine (3TC) in vitro and in patients treated with 3TC monotherapy. It was of interest to determine the fidelity of RNA-dependent DNA polymerization (RDDP) of M184V RT compared with wild-type because this step occurs first in reverse transcription; errors made during this step may be copied in subsequent polymerization steps. Using an in vitro mispaired primer extension assay, M184V-mutated RT exhibited 3-49-fold decreased frequency of mispair extension compared with wild-type RT. Fidelity differences between M184V and wild type RT were most marked in extension of A:G (49-fold) and A:C (16-fold) mispairs, with only a marginal (3-fold) decrease in the extension of A:A mispairs. RT containing a methionine to isoleucine (M184I) mutation showed only slight increases in RDDP fidelity compared with wild-type, ranging from 1.5- to 6 fold increases. Of the three RTs tested, wild-type RT was the most error-prone, with mispair extension frequencies ranging from 6.674 x 10(-1) to 7.454 x10(-2). PMID- 9358164 TI - The RNA moiety of chick embryo 5-methylcytosine- DNA glycosylase targets DNA demethylation. AB - We have previously shown that DNA demethylation by chick embryo 5-methylcytosine (5-MeC)-DNA glycosylase needs both protein and RNA. RNA from enzyme purified by SDS-PAGE was isolated and cloned. The clones have an insert ranging from 240 to 670 bp and contained on average one CpG per 14 bases. All six clones tested had different sequences and did not have any sequence homology with any other known RNA. RNase-inactivated 5-MeC-DNA glycosylase regained enzyme activity when incubated with recombinant RNA. However, when recombinant RNA was incubated with the DNA substrate alone there was no demethylation activity. Short sequences complementary to the labeled DNA substrate are present in the recombinant RNA. Small synthetic oligoribonucleotides (11 bases long) complementary to the region of methylated CpGs of the hemimethylated double-stranded DNA substrate restore the activity of the RNase-inactivated 5-MeC-DNA glycosylase. The corresponding oligodeoxyribonucleotide or the oligoribonucleotide complementary to the non methylated strand of the same DNA substrate are inactive when incubated in the complementation test. A minimum of 4 bases complementary to the CpG target sequence are necessary for reactivation of RNase-treated 5-MeC-DNA glycosylase. Complementation with double-stranded oligoribonucleotides does not restore 5-MeC DNA glycosylase activity. An excess of targeting oligoribonucleotides cannot change the preferential substrate specificity of the enzyme for hemimethylated double-stranded DNA. PMID- 9358163 TI - High affinity binding of MEF-2C correlates with DNA bending. AB - To regulate lineage-specific gene expression in many cell types, members of the myocyte enhancer factor-2 (MEF-2) family of transcription factors cooperate with basic helix-loop-helix (bHLH) proteins, which show only limited intrinsic DNA binding specificity. We investigated the DNA binding properties of MEF-2C in vitro and show that the inherent bendability of the MEF site is one of the principal structural characteristics recognized by MEF-2C. Measurements of the apparent dissociation constants of MEF-2C complexes with several DNA sequences revealed that MEF-2C bound with high affinity to DNA sequences containing a MEF site. Mutations in the MEF site which did not affect the bendability of the DNA changed the free energy of binding only marginally. However, reducing the intrinsic bendability of the DNA binding site through an AA-->GC substitution increased the half-maximal binding concentration of MEF-2C by almost one order of magnitude. Electrophoretic mobility shift assays revealed markedly reduced MEF-2C binding to DNA containing 2,6-diaminopurine. On binding to MEF-2C the maximum ellipticity at 275 nm in the CD spectrum of DNA containing a MEF site was red shifted by 4 nm and its intensity reduced significantly, while a slight blue shift of <1 nm was observed for a mutant DNA sequence with reduced bendability (AA-->GC). Bending analysis by circular permutation assay revealed that the DNA in the cognate complex was bent by 49 degrees , while the DNA in the complex with the mutant oligonucleotide was largely unbent. PMID- 9358165 TI - Structural studies on tRNA acceptor stem microhelices: exchange of the discriminator base A73 for G in human tRNALeu switches the acceptor specificity from leucine to serine possibly by decreasing the stability of the terminal G1 C72 base pair. AB - Correct recognition of transfer RNAs (tRNAs) by aminoacyl-tRNA synthetases (aaRS) is crucial to the maintenance of translational fidelity. The discriminator base A73 in human tRNALeuis critical for its specific recognition by the aaRS. Exchanging A73 for G abolishes leucine acceptance and converts it into a serine acceptor in vitro . Two RNA microhelices of 24 nt length that correspond to the tRNALeuacceptor stem and differ only in the discriminator base were synthesized: a wild-type tRNALeumicrohelix, where nt 21 corresponds to the discriminator base position 73, and an A21G mutant microhelix. To investigate whether different identities of both tRNAs are caused by conformational differences, NMR and UV melting experiments were performed on both microhelices. Two-dimentional NOESY spectra showed both microhelices to exhibit the same overall conformation at their 3'-CCA ends. Thermodynamic analysis and melting behaviour of the base paired imino protons observed by NMR spectroscopy suggest that the A21G (A73G in tRNA) exchange results in a decrease of melting transition cooperativity and a destabilization of the terminal G1-C20 (G1-C72 in tRNA) base pair. Furthermore, the fact that this 3'-terminal imino proton is more solvent-exposed at physiological temperature might be another indication for the importance of the stability of the terminal base pair for specific tRNA recognition. PMID- 9358166 TI - The yeast 8-oxoguanine DNA glycosylase (Ogg1) contains a DNA deoxyribophosphodiesterase (dRpase) activity. AB - The yeast OGG1 gene was recently cloned and shown to encode a protein that possesses N-glycosylase/AP lyase activities for the repair of oxidatively damaged DNA at sites of 7,8-dihydro-8-oxoguanine (8-oxoguanine). Similar activities have been identified for Escherichia coli formamidopyrimidine-DNA glycosylase (Fpg) and Drosophila ribosomal protein S3. Both Fpg and S3 also contain a deoxyribophosphodiesterase (dRpase) activity that removes 2-deoxyribose-5 phosphate at an incised 5' apurinic/apyrimidinic (AP) sites via a beta elimination reaction. Drosophila S3 also has an additional activity that removes trans-4-hydroxy-2-pentenal-5-phosphate at a 3' incised AP site by a Mg2+ dependent hydrolytic mechanism. In view of the substrate similarities between Ogg1, Fpg and S3 at the level of base excision repair, we examined whether Ogg1 also contains dRpase activities. A glutathione S-transferase fusion protein of Ogg1 was purified and subsequently found to efficiently remove sugar-phosphate residues at incised 5' AP sites. Activity was also detected for the Mg2+ dependent removal of trans -4-hydroxy-2-pentenal-5-phosphate at 3' incised AP sites and from intact AP sites. Previous studies have shown that DNA repair proteins that possess AP lyase activity leave an inefficient DNA terminus for subsequent DNA synthesis steps associated with base excision repair. However, the results presented here suggest that in the presence of MgCl2, Ogg1 can efficiently process 8-oxoguanine so as to leave a one nucleotide gap that can be readily filled in by a DNA polymerase, and importantly, does not therefore require additional enzymes to process trans -4-hydroxy-2-pentenal-5-phosphate left at a 3' terminus created by a beta-elimination catalyst. PMID- 9358167 TI - Placement of the alpha-sarcin loop within the 50S subunit: evidence derived using a photolabile oligodeoxynucleotide probe. AB - We report the synthesis of a radioactive, photolabile oligodeoxyribonucleotide probe and its exploitation in identifying 50S ribosomal subunit components neighboring the alpha-sarcin loop. The probe is complementary to 23S rRNA nt 2653 2674. Photolysis of the complex formed between the probe and 50S subunits leads to site-specific probe photoincorporation into proteins L2, the most highly labeled protein, L1, L15, L16 and L27, labeled to intermediate extents, and L5, L9, L17 and L24, each labeled to a minor extent. Portions of each of these proteins thus lie within 23 A of nt U2653. These results lead us to conclude that the alpha-sarcin loop is located at the base of the L1 projection within the 50S subunit. Such placement, near the peptidyl transferase center, provides a rationale for the extreme sensitivity of ribosomal function to cleavage of the alpha-sarcin loop. PMID- 9358168 TI - RAGA: RNA sequence alignment by genetic algorithm. AB - We describe a new approach for accurately aligning two homologous RNA sequences when the secondary structure of one of them is known. To do so we developed two software packages, called RAGA and PRAGA, which use a genetic algorithm approach to optimize the alignments. RAGA is mainly an extension of SAGA, an earlier package for multiple protein sequence alignment. In PRAGA several genetic algorithms run in parallel and exchange individual solutions. This method allows us to optimize an objective function that describes the quality of a RNA pairwise alignment, taking into account both primary and secondary structure, including pseudoknots. We report results obtained using PRAGA on nine test cases of pairs of eukaryotic small subunit rRNA sequence (nuclear and mitochondrial). PMID- 9358170 TI - Solution structure of RNA duplexes containing alternating CG base pairs: NMR study of r(CGCGCG)2 and 2'-O-Me(CGCGCG)2 under low salt conditions. AB - Structures of r(CGCGCG)2 and 2'-O-Me(CGCGCG)2 have been determined by NMR spectroscopy under low salt conditions. All protons and phosphorus nuclei resonances have been assigned. Signals of H5'/5" have been assigned stereospecifically. All 3JH,H and 3JP,H coupling constants have been measured. The structures were determined and refined using an iterative relaxation matrix procedure (IRMA) and the restrained MD simulation. Both duplexes form half-turn, right-handed helices with several conformational features which deviate significantly from a canonical A-RNA structure. Duplexes are characterised as having C3'-endo sugar pucker, very low base-pair rise and high helical twist and inclination angles. Helices are overwound with <10 bp per turn. There is limited inter-strand guanine stacking for CG steps. Within CG steps of both duplexes, the planes of the inter-strand cytosines are not parallel while guanines are almost parallel. For the GC steps this pattern is reversed. The 2'-O-methyl groups are spatially close to the 5'-hydrogens of neighbouring residues from the 3'-side and are directed towards the minor groove of 2'-O-Me(CGCGCG)2 forming a hydrophobic layer. Solution structures of both duplexes are similar; the effect of 2'-O methylation on the parent RNA structure is small. This suggests that intrinsic properties imposed by alternating CG base pairs govern the overall conformation of both duplexes. PMID- 9358169 TI - 2'-Fluoro modified nucleic acids: polymerase-directed synthesis, properties and stability to analysis by matrix-assisted laser desorption/ionization mass spectrometry. AB - Fragmentation is a major factor limiting mass range and resolution in the analysis of DNA by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Protonation of the nucleobase leads to base loss and backbone cleavage by a mechanism similar to the depurination reactions employed in the chemical degradation method of DNA sequencing. In a previous study [Tang,W., Zhu,L. and Smith,L.M. (1997) Anal. Chem ., 69, 302-312], the stabilizing effect of substituting the 24 hydrogen with an electronegative group such as hydroxyl or fluorine was investigated. These 24 substitutions stabilized the N-glycosidic linkage, blocking base loss and subsequent backbone cleavage. For such chemical modifications to be of practical significance, it would be useful to be able to employ the corresponding 24-modified nucleoside triphosphates in the polymerase directed synthesis of DNA. This would provide an avenue to the preparation of 24 modified PCR fragments and dideoxy sequencing ladders stabilized for MALDI analysis. In this paper methods are described for the polymerase-directed synthesis of 24-fluoro modified DNA, using commercially available 24 fluoronucleoside triphosphates. The ability of a number of DNA and RNA polymerases to incorporate the 24-fluoro analogs was tested. Four thermostable DNA polymerases [Pfu (exo-), Vent (exo-), Deep Vent (exo-) and UlTma] were found that were able to incorporate 24-fluoronucleotides with reasonable efficiency. In order to perform Sanger sequencing reactions, the enzymes' ability to incorporate dideoxy terminators in conjunction with the 24-fluoronucleotides was evaluated. UlTma DNA polymerase was found to be the best of the enzymes tested for this purpose. MALDI analysis of enzymatically produced 24-fluoro modified DNA using the matrix 2,5-dihydroxy benzoic acid showed no base loss or backbone fragmentation, in contrast to the extensive fragmentation evident with unmodified DNA of the same sequence. PMID- 9358171 TI - Crystal structure of 2'-O-Me(CGCGCG)2, an RNA duplex at 1.30 A resolution. Hydration pattern of 2'-O-methylated RNA. AB - The molecular and crystal structure of 2'-O-Me (CGCGCG)2 has been determined using synchrotron radiation at near-atomic resolution (1.30 A), the highest resolution to date in the RNA field. The crystal structure is a half-turn A-type helix with some helical parameters deviating from canonical A-RNA, such as low base pair rise, elevated helical twist and inclination angles. In CG steps, inter strand guanines are parallel while cytosines are not parallel. In steps GC this motif is reversed. This type of regularity is not seen in other RNA crystal structures. The structure includes 44 water molecules and two hydrated Mg2+ions one of which lies exactly on the crystallographic 2-fold axis. There are distinct patterns of hydration in the major and the minor grooves. The major groove is stabilised by water clusters consisting of fused five- and six-membered rings. Minor groove contains only a single row of water molecules; each water bridges either two self-parallel cytosines or two self-parallel guanines by a pair of hydrogen bonds. The structure provides the first view of the hydration scheme of 2'-O-methylated RNA duplex. PMID- 9358172 TI - Opening of the extraordinarily stable mini-hairpin d(GCGAAGC). AB - For the purposes of the antisense strategy oligodeoxyribonucleotides can be protected against serum and cell nuclease digestion by tagging at their 3'-end with a sequence naturally forming a very stable hairpin, d(GCGAAGC). This nuclease-resistant hairpin is also known for its high thermostability. We demonstrate in this study that attachment of d(GCGAAGC) at the 3'-end of an oligodeoxyribonucleotide does not hinder hybridization of the 5'-part of this oligonucleotide to a complementary DNA strand. Moreover, the hairpin is in equilibrium between a folded and an open structure, with an energy minimum in favor of pairing if it is possible, even with mismatches. PMID- 9358173 TI - Deoxyribonucleotide-containing RNAs: a novel class of templates for HIV-1 reverse transcriptase. AB - Deoxyribonucleotide-containing RNA-like polynucleotides (dcRNAs) were synthesized by mutant T7 RNA polymerase and their structures confirmed by sequencing. dcRNAs annealed with a 20mer oligodeoxyribonucleotide primer were tested as templates/primers in the reverse transcription reaction catalyzed by HIV-1 reverse transcriptase (RT). All dcRNAs were shown to be efficient templates for both wild-type RT and RT mutants, containing 'AZT-resistant' mutations. Differences in the patterns of the DNA products of RNA- and dcRNA-driven reverse transcription were demonstrated. The kinetic characteristics for dcRNAs utilization were compared with the corresponding parameters for RNA/DNA and DNA/DNA templates/primers. The respective K m values for dcRNAs appear to be intermediate between those for RNA and DNA templates. A correlation equation connecting apparent K m value for template/primer and the number of deoxyribonucleotide substitutions in RNA template is proposed. PMID- 9358174 TI - Analysis of the yeast genome: identification of new non-coding and small ORF containing RNAs. AB - The genome sequences from increasing numbers of organisms allow for rapid and organized examination of gene expression. Yet current computational-based paradigms for gene recognition are limited and likely to miss genes expressing non-coding RNAs or mRNAs with small open reading frames (ORFs). We have utilized two strategies to determine if there are additional transcripts in the yeast Saccharomyces cerevisiae that were not identified in previous analyses of the genome. In one approach, we identified strong consensus polymerase III promoters based on sequence, and determined experimentally if these promoters drive the expression of an RNA polymerase III transcript. This approach led to the identification of a new, non-essential 170 nt non-coding RNA. An alternative strategy analyzed RNA expression from large sequence gaps>2 kb between predicted ORFs. Fifteen unique RNA transcripts ranging in size from 161 to 1200 nt were identified from a total of 59 sequence gaps. Several of these RNAs contain unusually small potential ORFs, while one is clearly non-coding and appears to be a small nucleolar RNA. These results suggest that there are likely to be additional previously unidentified non-coding RNAs in yeast, and that new paradigms for gene recognition will be required to identify all expressed genes from an organism. PMID- 9358175 TI - Statistical modeling and analysis of the LAGLIDADG family of site-specific endonucleases and identification of an intein that encodes a site-specific endonuclease of the HNH family. AB - The LAGLIDADG and HNH families of site-specific DNA endonucleases encoded by viruses, bacteriophages as well as archaeal, eucaryotic nuclear and organellar genomes are characterized by the sequence motifs 'LAGLIDADG' and 'HNH', respectively. These endonucleases have been shown to occur in different environments: LAGLIDADG endonucleases are found in inteins, archaeal and group I introns and as free standing open reading frames (ORFs); HNH endonucleases occur in group I and group II introns and as ORFs. Here, statistical models (hidden Markov models, HMMs) that encompass both the conserved motifs and more variable regions of these families have been created and employed to characterize known and potential new family members. A number of new, putative LAGLIDADG and HNH endonucleases have been identified including an intein-encoded HNH sequence. Analysis of an HMM-generated multiple alignment of 130 LAGLIDADG family members and the three-dimensional structure of the I- Cre I endonuclease has enabled definition of the core elements of the repeated domain (approximately 90 residues) that is present in this family of proteins. A conserved negatively charged residue is proposed to be involved in catalysis. Phylogenetic analysis of the two families indicates a lack of exchange of endonucleases between different mobile elements (environments) and between hosts from different phylogenetic kingdoms. However, there does appear to have been considerable exchange of endonuclease domains amongst elements of the same type. Such events are suggested to be important for the formation of elements of new specficity. PMID- 9358176 TI - Increased DNA binding and sequence discrimination of PNA oligomers containing 2,6 diaminopurine. AB - The synthesis of a diaminopurine PNA monomer, N-[N6-(benzyloxycarbonyl)-2,6 diaminopurine-9-yl] acetyl-N-(2-t-butyloxycarbonylaminoethyl)glycine, and the incorporation of this monomer into PNA oligomers are described. Substitution of adenine by diaminopurine in PNA oligomers increased the T m of duplexes formed with complementary DNA, RNA or PNA by 2.5-6.5 degrees C per diaminopurine. Furthermore, discrimination against mismatches facing the diaminopurine in the hybridizing oligomer is improved. Finally, a homopurine decamer PNA containing six diaminopurines is shown to form a (gel shift) stable strand displacement complex with a target in a 246 bp double-stranded DNA fragment. PMID- 9358177 TI - Relative stability of triplexes containing different numbers of T.AT and C+.GC triplets. AB - We have used DNase I footprinting to compare the stability of parallel triple helices containing different numbers of T.AT and C+. GC triplets. We have targeted a fragment containing the 17mer sequence 5'-AGGAAGAGAAAAAAGAA with the 9mer oligonucleotides 5'-TCCTTCTCT, 5'-TTCTCTTTT and 5'-TTTTTTCTT, which form triplexes at the 5'-end, centre and 3'-end of the target site respectively. Quantitative DNase I footprinting has shown that at pH 5.0 the dissociation constants of these oligonucleotides are 0.13, 4.7 and >30 microM respectively, revealing that increasing the proportion of C+.GC triplets increases triplex stability. The results suggest that the positive charge on the protonated cytosine contributes to triplex stability, either by a favourable interaction with the stacked pisystem or by screening the charge on the phosphate groups. In the presence of a naphthylquinoline triplex binding ligand all three oligonucleotides bind with similar affinities. At pH 6.0 these triplexes only form in the presence of the triplex binding ligand, while at pH 7.5 footprints are only seen with the oligonucleotide which generates the fewest number of C+.GC triplets (TTTTTTCTT) in the presence of the ligand. PMID- 9358178 TI - Non-homologous DNA end joining in plant cells is associated with deletions and filler DNA insertions. AB - Double strand DNA breaks in plants are primarily repaired via non-homologous end joining. However, little is known about the molecular events underlying this process. We have studied non-homologous end joining of linearized plasmid DNA with different termini configurations following transformation into tobacco cells. A variety of sequences were found at novel end junctions. Joining with no sequence alterations was rare. In most cases, deletions were found at both ends, and rejoining usually occurred at short repeats. A distinct feature of plant junctions was the presence of relatively large, up to 1.2 kb long, insertions (filler DNA), in approximately 30% of the analyzed clones. The filler DNA originated either from internal regions of the plasmid or from tobacco genomic DNA. Some insertions had a complex structure consisting of several reshuffled plasmid-related regions. These data suggest that double strand break repair in plants involves extensive end degradation, DNA synthesis following invasion of ectopic templates and multiple template switches. Such a mechanism is reminiscent of the synthesis-dependent recombination in bacteriophage T4. It can also explain the frequent 'DNA scrambling' associated with illegitimate recombination in plants. PMID- 9358179 TI - Both the polypyrimidine tract and the 3' splice site function prior to the first step of splicing in fission yeast. AB - While it is known that several trans -acting splicing factors are highly conserved between Schizosaccharomyces pombe and mammals, the roles of cis -acting signals have received comparatively little attention. In Saccharomyces cerevisiae, sequences downstream from the branch point are not required prior to the first transesterification reaction, whereas in mammals the polypyrimidine tract and, in some introns, the 3' AG dinucleotide are critical for initial recognition of an intron. We have investigated the contribution of these two sequence elements to splicing in S.pombe. To determine the stage at which the polypyrimidine tract functions, we analyzed the second intron of the cdc2 gene (cdc 2-Int2), in which pyrimidines span the entire interval between the branch point and 3' splice site. Our data indicate that substitution of a polypurine tract results in accumulation of linear pre-mRNA, while expanding the polypyrimidine tract enhances splicing efficiency, as in mammals. To examine the role of the AG dinucleotide in cdc 2-Int2 splicing, we mutated the 3' splice junction in both the wild-type and pyrimidine tract variant RNAs. These changes block the first transesterification reaction, as in a subset of mammalian introns. However, in contrast to the situation in mammals, we were unable to rescue the first step of splicing in a 3' splice site mutant by expanding the polypyrimidine tract. Mutating the terminal G in the third intron of the nda 3 gene (nda 3-Int3) also blocks the first transesterification reaction, suggesting that early recognition of the 3' splice site is a general property of fission yeast introns. Counter to earlier work with an artificial intron, it is not possible to restore the first step of splicing in cdc 2-Int2 and nda 3-Int3 3' splice site mutants by introducing compensatory changes in U1 snRNA. These results highlight the diversity and probable redundancy of mechanisms for identifying the 3' ends of introns. PMID- 9358180 TI - Baculovirus-mediated expression and characterization of the full-length murine DNA methyltransferase. AB - The original cDNA sequence reported for the murine DNA methyltransferase (MTase) was not full length. Recently, additional cDNA sequences have been reported that lie upstream of the original and contain an extended open reading frame with three additional ATGs in frame with the coding region [Tucker et al . (1996) Proc. Natl. Acad. Sci. USA , 93, 12920-12925; Yoder et al . (1996) J. Biol. Chem . 271, 31092-31097]. Genomic DNA upstream of this ATG contains two more ATGs in frame and no obvious splice site. We have constructed, and expressed in baculovirus, MTase clones that begin at each of these four ATGs and examined their properties. Constructs beginning with any of the first three ATGs as their initiator methionines give a predominant DNA MTase band of approximately 185 kDa on SDS-PAGE corresponding to translational initiation at the third ATG. The fourth ATG construct gives a much smaller protein band of 173 kDa. The 185 kDa protein was purified by HPLC, characterized by mass spectrometry and has a measured molecular mass of 184 +/- 0.5 kDa. All of these MTases were functional in vitro and steady state kinetic analysis showed that the recombinant proteins exhibit similar kinetic properties irrespective of their length. The homogeneous recombinant enzyme from the fourth ATG construct shows a 2.5-fold preference for a hemi-methylated DNA substrate as compared to an unmethylated substrate, whereas the 185 kDa protein is equally active on both substrates. The kinetic properties of the recombinant enzyme are similar to those reported for the native MTase derived from murine erythroleukemia cells. The new clones are capable of yielding large quantities of intact MTases for further structural and functional studies. PMID- 9358181 TI - Peptide nucleic acid-anthraquinone conjugates: strand invasion and photoinduced cleavage of duplex DNA. AB - A bis-peptide nucleic acid (PNA)-anthraquinone imide (AQI) conjugate has been synthesized and shown to form strand invasion complexes with a duplex DNA target. The two arms of the bis-PNA each consist of five consecutive thymine residues and are linked by a flexible, hydrophilic spacer. Probing with potassium permanganate reveals that the bis-PNA complexes to duplex DNA at A5.T5sites with local displacement of the T5DNA strand. The 5 bp sequence targeted by the PNA is the shortest strand invasion complex reported to date. Irradiation of the strand invasion complex results in asymmetric cleavage of the displaced strand, with more efficient cleavage at the 3'-end of the loop. This result indicates that the bis-PNA binds to the DNA such that the C-terminal T5sequence forms the strand invasion complex, leaving the N-terminal T5sequence to bind by triplex formation, thereby placing the AQI closer to the 3'-end of the displaced strand, consistent with the observed photocleavage pattern. The ability of the PNA to directly report its binding site by photoinduced cleavage could have significant utility in mapping the secondary and tertiary structure of nucleic acids. PMID- 9358182 TI - RnaViz, a program for the visualisation of RNA secondary structure. AB - RnaViz is a user-friendly, portable, windows-type program for producing publication-quality secondary structure drawings of RNA molecules. Drawings can be created starting from DCSE alignment files if they incorporate structure information or from mfold ct files. The layout of a structure can be changed easily. Display of special structural elements such as pseudo-knots or unformatted areas is possible. Sequences can be automatically numbered, and several other types of labels can be used to annotate particular bases or areas. Although the program does not try to produce an initially non-overlapping drawing, the layout of a properly positioned structure drawing can be applied to a newly created drawing using skeleton files. In this way a range of similar structures can be drawn with a minimum of effort. Skeletons for several types of RNA molecule are included with the program. PMID- 9358183 TI - Exploiting unassigned codons in Micrococcus luteus for tRNA-based amino acid mutagenesis. AB - An alternative to suppression of stop codons for the biosynthetic insertion of non-natural amino acids has been developed. Micrococcus luteus , a Gram-positive bacterium, is incapable of translating at least two codons. One of these unused codons was inserted in a gene to act as a nonsense site. An aminoacylated tRNA was synthesized which was complementary to this codon. The gene containing the missing codon was expressed in vitro in a M.luteus transcription/translation system. Read-through of the missing codon occurred only when the complementary tRNA was included. The results demonstrate that M.luteus can be used for incorporation of amino acids via synthetically prepared aminoacylated tRNAs. The use of a M. luteus translation system provides a method for incorporation of non natural amino acids which avoids the use of stop codons. PMID- 9358184 TI - Construction of a semisynthetic antibody library using trinucleotide oligos. AB - A semisynthetic antibody library composed of single chain Fv fragments (scFv) was constructed by replacing the heavy chain CDR3 region of a human scFv by a random sequence of eight amino acids using trinucleotide codons. After cloning into a phage display vector, an antibody library was generated with a complexity of 8 x 10(8) independent clones. The library was screened for binders to dinitrophenol, fluorescein isothiocyanate and 3-nitro-4-hydroxy-5-iodophenylacetic acid. scFv antibodies that specifically bound the antigen were obtained in each case. PMID- 9358185 TI - Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. AB - A very simple, fast, universally applicable and reproducible method to extract high quality megabase genomic DNA from different organisms is described. We applied the same method to extract high quality complex genomic DNA from different tissues (wheat, barley, potato, beans, pear and almond leaves as well as fungi, insects and shrimps' fresh tissue) without any modification. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low technology laboratories. The amount of tissue required by this method is approximately 50-100 mg. The quantity and the quality of the DNA extracted by this method is high enough to perform hundreds of PCR based reactions and also to be used in other DNA manipulation techniques such as restriction digestion, Southern blot and cloning. PMID- 9358186 TI - Adaptor-tagged competitive PCR: a novel method for measuring relative gene expression. AB - A simple and reliable PCR-based method to quantitate gene expression is described. Following the digestion of double-stranded cDNA by a restriction enzyme, an adaptor is ligated to a cDNA from a first RNA sample, and another adaptor to a second RNA sample. The two adaptors share a common sequence at the outer region, but differ in size. Equal amounts of the ligated samples are mixed, and amplified by an adaptor-primer and a primer specific to the gene of interest. Products derived from the two sources differ in size, and can be separated by denaturing polyacrylamide gel electrophoresis. The ratio of the two products reveals the relative level of gene expression. Since the technique avoids the need to construct internal standards, it is especially useful for the analysis of many different gene transcripts. PMID- 9358187 TI - Monitoring the amplification of CATCH, a 3SR based cooperatively coupled isothermal amplification system, by fluorimetric methods. AB - Three different types of fluorescence detection methods were employed to monitor amplification of a previously established isothermal cooperatively coupled amplification system as it can serve as a tool for the investigation of fundamental issues in evolutionary optimization. By using 5'IRD-41 fluorescent labeled primers, the intercalating dye TOPRO-1 and a 5'fluorescin/3'DABCYL 4-(4 dimethylamino-phenylazo)benzoic acid labeled ss 24 nt DNA, evolving molecular cooperation is accessible, sequence specifically as well as non-sequence specifically without using radioactivity. PMID- 9358188 TI - Isolation of nuclei for chromatin analysis in fission yeast. AB - The methods available for analysis of the chromatin of Schizosaccharomyces pombe are time consuming (>8 h) and/or result in some degradation of the chromatin. Here we report an optimised method for the preparation of spheroplasts and the isolation of nuclei which takes <25 min and is suitable for analysis of chromatin structure by micrococcal nuclease, restriction endonuclease or by immunoprecipitation. PMID- 9358189 TI - Pharmacokinetics of continuous-release carbidopa/levodopa. AB - This article reviews the pharmacokinetics of Sinemet CR, a controlled-release (CR) levodopa preparation. The main influences on the kinetic profile are as follows: absorption, which depends on the dissolution characteristics of the tablet, the pattern of gastric emptying, and the rate of uptake in the small intestine; distribution, which is determined by rates of levodopa transport from gut to blood and from blood to brain; and biotransformation, which is affected peripherally by L-aromatic amino acid decarboxylase (LAAAD) and catechol-O-methyl transferase (COMT), and centrally by LAAAD, COMT, and monoamine oxidase. The kinetics of Sinemet CR are limited by rates of absorption (which depend on the dose administered), the conformation of the tablet, and daily variations in the patterns of gastric emptying (influenced by the presence or absence of food). Levodopa must also compete with food-derived amino acids for transport across the gut-blood and blood-brain barriers; this competition effectively limits the drug's rate of distribution. Finally, biotransformation is limited by the activity of LAAAD and COMT in the periphery. Plasma profiles of levodopa after administration of Sinemet CR can vary, depending on the age of the patient and the time of day when the drug is administered. Nevertheless, the pharmacokinetic profile of the preparation has a number of advantages over that of Sinemet, in that it offers a steadier climb to peak plasma concentrations that are less extreme and of greater duration. PMID- 9358191 TI - Early idiopathic parkinsonism: initiation and optimization of treatment. AB - Once a diagnosis of idiopathic parkinsonism has been made, the choice and timing of therapy depend almost entirely on the patient's need for symptomatic relief, as no presently available therapy has any effect on the pathogenesis of the disease. Five categories of drugs are available for the treatment of idiopathic parkinsonism. Anticholinergic agents are effective against tremor but have prominent adverse effects. Amantadine has similar effects but is more active against rigidity and bradykinesia. Selegiline is a monoamine oxidase-B inhibitor; once thought to affect the pathogenesis of idiopathic parkinsonism, it is now known to offer only symptomatic relief. The dopamine agonists (bromocriptine, pergolide, and lisuride) stimulate D2 receptors; they have antiparkinsonian effects and tolerance profiles broadly similar to those of levodopa but are slightly less efficacious. Pleural effusions and pulmonary fibrosis are unusual but important complications of these drugs; chest x-ray examinations are therefore recommended for all patients starting such treatment. Levodopa (combined with an extracerebral decarboxylase inhibitor to prevent nausea, the main adverse effect) has become the standard antiparkinsonism treatment. Patients using this preparation can suffer considerable variations in mobility and dyskinesia, which may be related to rapid, large-scale oscillations in plasma levodopa concentrations. Controlled-release (CR) preparations have been developed in an attempt to minimize these fluctuations and reduce long-term side effects. There is no universally agreed treatment for idiopathic parkinsonism. However, experience shows that a good balance of antiparkinsonian activity and adverse effects can be obtained by initiating treatment with a combination of levodopa and a decarboxylase inhibitor. A dopamine agonist can be added if the disease progresses and increased therapeutic activity is required. PMID- 9358190 TI - Clinical implications of sustained dopaminergic stimulation. AB - Fluctuations in motor performance are the major problems in chronic management of Parkinson's disease. Most of these fluctuations reflect the decline of levodopa availability. As a consequence, levodopa dosage might be increased and the interdose interval progressively shortened. The postsynaptic dopamine receptors at this point are exposed to a nonphysiologic shift in dopamine level, which may induce changes at the receptor site and contribute to the appearance of "on-off" phenomena and dyskinesias. We compared a group of 18 patients treated for 60 consecutive months with continuous subcutaneous lisuride infusion with a group of 20 patients treated with conventional oral levodopa treatment. The clinical evaluations performed during the study showed in the lisuride group only a worsening of dyskinesias, whereas the other symptoms remained unchanged. In the other group the evaluation scores showed a significant worsening of all long-term treatment complications. The slow-release preparations of levodopa may ensure a more continuous dopaminergic stimulation than standard formulations. However, the use of these compounds is difficult in severely fluctuating patients because the lack of a plasma peak level usually leads to a very long delay before patients turn "on." We studied the pharmacokinetic and clinical effects of the two slow release preparations of levodopa [Madopar HBS and Sinemet controlled-release (CR)] and a combination of Sinemet CR plus standard Sinemet in 13 fluctuating parkinsonian patients. The results of this study show that the combination of standard Sinemet and Sinemet CR ensures a more prolonged clinical effect with a very short latency to the "on" phase. PMID- 9358192 TI - Mid-stage parkinsonism with mild motor fluctuations. AB - Most patients with Parkinson's disease who undergo levodopa therapy eventually develop fluctuations in the diurnal control of their symptoms. These fluctuations, when mild, consist mostly of the "wearing-off" effect--the re emergence of symptoms several hours after the ingestion of each dose of levodopa. In some patients, "wearing-off" in the daytime is preceded by, or associated with, the development of nocturnal and early morning akinesia. Some patients at this stage also have mild (mostly peak dose or square-wave) choreoathetoid dyskinesias. In patients with "wearing-off," "on" periods tend to occur when plasma levels of levodopa are high; nocturnal and early morning akinesia and diurnal "off" periods tend to occur at times when plasma levodopa levels are low, reflecting insufficient concentrations of dopamine in the striatum. Treatment strategies in patients with mid-stage parkinsonism and mild fluctuations are primarily directed toward enhancing dopaminergic transmission in the striatum. This is achieved by increasing the supply of levodopa to the brain or by administering drugs that either prolong the effect of synaptic dopamine from levodopa, or activate postsynaptic dopamine receptors. This presentation will discuss the advantages and possible drawbacks of these different strategies, which include the use of controlled-release levodopa preparations, deprenyl, and the new monoamine oxidase (MAO)-B inhibitors, catechol-O-methyl transferase (COMT) inhibitors, and the various "direct" dopamine agonists. PMID- 9358193 TI - Problems with long-term levodopa therapy for Parkinson's disease. AB - The introduction of levodopa 25 years ago revolutionized the management of Parkinson's disease. However, it soon became apparent that the drug offered only symptomatic relief and did not affect the underlying pathology. Moreover, chronic use of the drug was associated with a range of adverse effects. Current therapeutic strategies seek to delay long-term complications of treatment for as long as possible. However, once they appear, most adverse effects are amenable to some form of management. A number of therapeutic strategies are available for treatment of Parkinson's disease. The final choice of therapy depends on the individual circumstances and requirements of the patient and should balance tolerance for adverse effects with the amount of symptomatic relief required. Patients receiving long-term levodopa therapy must contend with some adverse effects. After 5 years the majority of these patients suffer fluctuations, dyskinesias, toxicity, or loss of efficacy. Fluctuations can be reduced by changing the drug regimen to a combination therapy of Sinemet and Sinemet controlled-release (CR), or by the addition of deprenyl or a dopamine agonist. Variations in gastric emptying and absorption of levodopa and dietary factors become important. Dyskinesias in long-term levodopa therapy are poorly understood and difficult to manage, although dopamine agonists can be of some use. As the disease progresses, new disabilities appear that are less responsive to levodopa, and its efficacy can appear to diminish, with increased doses often leading to toxicity. PMID- 9358194 TI - Rationale for the prevention of disseminated Mycobacterium avium-intracellulare complex disease. AB - The survival rate in patients with AIDS who have CD4+ cell counts < 75 cells/microliter is increasing because of improved preventive and treatment strategies for opportunistic infections and also because of the efficacy of antiretroviral drug treatment. These patients are at high risk of developing disseminated Mycobacterium avium-intracellulare (MAC) disease, which decreases both quality of life and life expectancy. Measures aimed at preventing MAC contamination are largely ineffective in decreasing the incidence of disseminated MAC disease in patients with AIDS, because of the large natural reservoir of MAC. Chemoprophylaxis is superior to early bacteriological diagnosis as a preventive strategy, and it is preferable to wait for the appearance of symptoms of disseminated MAC disease before a curative treatment is initiated. Well-conducted studies of clarithromycin or rifabutin monotherapy as chemoprophylaxis have demonstrated a decrease in the incidence of disseminated MAC disease, as well as an increase in quality of life and survival. Clarithromycin, azithromycin and rifabutin have all been shown to be effective as prophylaxis against disseminated MAC disease. Although some combinations of drugs have been shown to be more effective than monotherapy in preventing disseminated MAC disease, these regimens are more costly and have less favourable tolerability profiles than single-agent treatment. In conclusion, chemoprophylaxis is the most effective preventive strategy against disseminated MAC disease and has been shown to improve quality of life and to decrease the risk of death associated with this disease in AIDS patients. PMID- 9358195 TI - Prevention strategies for Mycobacterium avium-intracellulare complex (MAC) infection. A review of recent studies in patients with AIDS. AB - In patients with AIDS, disseminated Mycobacterium avium-intracellulare complex (MAC) infection is a common bacterial infection and is associated with considerable morbidity and mortality. In placebo-controlled studies, rifabutin, clarithromycin and azithromycin (administered as single agents) have been shown to prevent the development of MAC bacteraemia in patients with advanced HIV disease. Clarithromycin also conferred a survival benefit over placebo, but this was not initially observed with either rifabutin or azithromycin. Subsequently, the efficacy of single agent therapy was compared with that of combination treatment as prophylaxis against the development of disseminated MAC. In the AIDS Clinical Trials Group (ACTG) 196/Community Programs for Clinical Research on AIDS (CPCRA) 009 study, clarithromycin monotherapy and clarithromycin and rifabutin combination therapy regimens were both more effective than rifabutin monotherapy in reducing the incidence of MAC bacteraemia. However, the combination regimen was generally not well tolerated. In the California Consortium Treatment Group (CCTG)/Multiple Opportunistic Prevention Prophylactic Strategy (MOPPS) study, azithromycin plus rifabutin was significantly more effective than either agent administered alone, and azithromycin was more effective than rifabutin. However, azithromycin (alone or in combination with rifabutin) caused frequent gastrointestinal adverse events. Emergence of resistance in those failing prophylaxis appeared to be higher with clarithromycin than with azithromycin or rifabutin. The use of the combination regimen of clarithromycin plus rifabutin did not reduce the selection of clarithromycin-resistant isolates. Several issues need to be considered in the choice of MAC prophylaxis for the individual patient. On the basis of efficacy and potential drug interactions with protease inhibitors, clarithromycin or azithromycin is preferred to rifabutin. However, rifabutin is less likely to result in the emergence of resistance than the macrolides, and is likely to prevent tuberculosis, whereas azithromycin and clarithromycin will prevent some bacterial infections. Combination therapy for prophylaxis is not indicated in most situations. PMID- 9358196 TI - Ancillary benefits of Mycobacterium avium-intracellulare complex prophylaxis with clarithromycin in HIV-infected patients. AB - Because of the significant morbidity and mortality associated with opportunistic infections, prophylaxis has become routine practice in the management of immunocompromised patients such as those with AIDS. Clarithromycin, an antimicrobial agent with a broad spectrum of activity against most common respiratory pathogens as well as many protozoa, has proven to be effective for both treatment and prophylaxis of Mycobacterium avium-intracellulare complex (MAC) infection in AIDS patients. Results of a large multinational placebo controlled study suggest that clarithromycin for MAC prophylaxis provides additional benefits. In this study, clarithromycin statistically significantly reduced the incidence of Pneumocystis carinii pneumonia (5.3% of clarithromycin recipients vs 10.0% of placebo recipients; p = 0.021), community-acquired pneumonia (7.1 vs 13.0%; p = 0.010), Giardia lamblia infection (0.9 vs 2.9%; p = 0.048), and neoplastic diseases (1.8 vs 4.1%; p = 0.010) in AIDS patients with CD4+ counts of < or = 100 cells/microliter. PMID- 9358197 TI - Prevention of the selection of clarithromycin-resistant Mycobacterium avium intracellulare complex. AB - The prevalence of clarithromycin-resistant mutants in untreated bacterial populations of Mycobacterium avium-intracellulare complex (MAC) has been demonstrated to be between 10(-7) and 10(-8) colony-forming units (CFUs) in the beige mouse model. Selection of these mutants occurred during clarithromycin monotherapy if treatment was initiated when the bacterial population size reached approximately 10(8) CFUs per spleen. Likewise, selection of clarithromycin resistant MAC was induced in AIDS patients during therapy with clarithromycin alone or in combination with drugs that were ineffective for the treatment or prevention of MAC infection. Because the emergence of clarithromycin resistance during preventive therapy was observed exclusively in AIDS patients with CD4+ cell counts < or = 25 cells/microliter, clarithromycin monotherapy can be recommended for the prevention of MAC infection in AIDS patients with CD4+ cell counts of > or = 50 cells/microliter. However, a clarithromycin-containing combination regimen is recommended for patients with CD4+ cell counts < 50 cells/microliter. Since preliminary animal experiments and clinical trials indicate that amikacin, ethambutol or rifabutin in combination with clarithromycin may prevent, or at least delay, the selection of clarithromycin resistant mutants, further preventive trials are urgently needed to confirm these observations. PMID- 9358198 TI - A comparison of the pharmacokinetics of meropenem after intravenous administration by injection over 2, 3 and 5 minutes. AB - The pharmacokinetics of meropenem were determined in 9 healthy volunteers after the administration of 1 g dose by injection over 2, 3 or 5 min. Peak plasma concentrations were not significantly different across the three rates of administration and, due to the finite time required for complete mixing of the blood in the central compartment, did not always occur at the end of the injection. Overall exposure to meropenem was unchanged by the more rapid rates of administration. Plasma clearance, terminal half-life and volume of distribution were virtually unchanged. Within 10 min after the start of the injection, the plasma concentrations from all three injections were very similar indicating that dosing over 2, 3 or 5 min would result in similar antimicrobial cover and, therefore, comparable efficacy. Comparison of the data derived from the three injections indicated that rapid administration of meropenem did not appreciably alter its disposition pharmacokinetics. Tolerability of meropenem was unchanged with the more rapid administration rate. PMID- 9358199 TI - Effect of LDL-apheresis on the pharmacokinetics of the lipophilic antilipidemic agent probucol. AB - The effect of LDL-apheresis on the pharmacokinetics of antilipidemic agents has not been evaluated thoroughly. In this study, we investigated the effect of LDL apheresis on the pharmacokinetics of probucol, a lipophilic antilipidemic agent, by studying its distribution and changes in the blood concentration of probucol after LDL-apheresis. The concentrations of lipoproteins were measured before and after LDL-apheresis in eight patients with familial hypercholesterolemia taking probucol. Concentrations of probucol in the various lipoprotein fractions and plasma were measured by HPLC. The serum concentrations of probucol before and after LDL-apheresis were 39.8 +/- 3.3 and 16.5 +/- 1.6 micrograms/ml, and the correlation coefficient between the changes in the serum probucol concentration and those in the serum cholesterol concentration before and after LDL-apheresis was significant (r = 0.73, P < 0.01). Changes in the probucol and cholesterol concentrations after LDL-apheresis were mainly found in the LDL fraction. The calculated reductions in the serum contents of probucol and cholesterol were similar to the contents of probucol and cholesterol in the irrigation fluid of the dextran sulfate column. These data suggest that changes of probucol concentration in plasma by LDL-apheresis are mainly due to reductions in the LDL fraction. PMID- 9358200 TI - Changes in cyclosporine A kinetics after experimental reduction of renal parenchyma. AB - Experimental chronic renal insufficiency (produced by 5/6 ablation of renal parenchyma) is associated with changes in the kinetics of oral (intragastric) cyclosporine A (CyA). Compared with animals with intact renal parenchyma, significantly lower levels of CyA are reached under these conditions. The factors responsible for reduced CyA availability under these conditions have not yet been identified. PMID- 9358202 TI - Excretion of tacrolimus glucuronides in human bile. AB - Tacrolimus is extensively metabolized by the cytochrome P-450 system. Hepatic metabolic phase I reactions of tacrolimus include mainly demethylation and/or hydroxylation. No valid data have been published on phase II pathways (glucuronide- or sulfo-conjugation). In order to investigate these pathways, different beta-glucuronidase/sulfatase enzyme preparations were used to hydrolyse the conjugates potentially present in human bile extracts. Two analytical methods were used: a non-specific method, MEIA, and a specific combined HPLC/MEIA method. The influence of the extraction pH was investigated. After beta-glucuronidase hydrolysis and extraction at pH 5, tacrolimus concentrations, obtained either from HPLC-MEIA or MEIA, always appeared significantly higher, suggesting the presence of glucuronides in the bile. When the extraction was performed at pH 1.5, only the HPLC-MEIA concentrations appeared higher after hydrolysis. MEIA concentrations obtained before and after hydrolysis were similar. These data are consistent with the fact that glucuronides are extracted at pH 1.5 but not at pH 5 and suggest first that, without hydrolysis, the extracted glucuronides are separated from the tacrolimus fraction in the HPLC-MEIA procedure, and second, that the glucuronides are cross-detected by the monoclonal antibody in the immunoassay. From these data, it is concluded that clues have been found, suggesting the presence in human bile of tacrolimus glucuronides, which cross react with the monoclonal antibody, provided they are extracted in the sample tested. PMID- 9358201 TI - Skin distribution of fipronil by microautoradiography following topical administration to the beagle dog. AB - To investigate the localisation of fipronil in dog skin, [14C]-fipronil was topically applied to a male beagle dog (spot-on administration) at the therapeutic dose of 10 mg/kg. By means of autohistoradiography, the radioactivity was precisely detected in the skin and appendages at various intervals after application. Radioactivity was predominantly observed within the stratum corneum, the viable epidermis, and in the pilo-sebaceous units (mainly in the sebaceous glands and epithelial layers). [14C]-fipronil was significantly detected in these structures up to 56 days post-treatment, in the application zone (neck) but also in the lumbar zone, thus indicating the mechanical displacement of fipronil. No radioactivity was detected in either the dermal or the hypodermal layers, confirming the low percutaneous passage of fipronil. PMID- 9358203 TI - The hepatic and duodenal activities of some drug metabolizing enzymes in chickens: influence of infection with Escherichia coli endotoxin and coccidiosis. AB - The activities of the drug metabolizing enzymes aminopyrine-N-demethylase, aniline hydroxylase and UDP-glucuronyl transferase, together with protein concentration, were measured in liver microsomes and duodenal mucosa from healthy chickens and chickens experimentally infected with Escherichia coli endotoxin, or naturally infected with Eimeria necatrix and Eimeria tenella (clinically classified as slight, moderate or severe infections). E. coli (2 mg/kg, 3 days) and severe coccidiosis significantly decreased the activities of the three enzymes in the liver and duodenum. However, infection by the E. coli endotoxin at lower doses (0.5 and 1 mg/kg, 3 days) and moderate or slight coccidial infections had no significant effect on the activities of these enzymes. Neither E. coli nor coccidial infections significantly affected the liver/body weight ratio. However, infection with the E. coli endotoxin (2 mg/kg, 3 days) and the moderate or severe coccidial infections significantly reduced microsomal protein concentration in the liver. PMID- 9358204 TI - Nonlinear binding of sex steroids to albumin and sex hormone binding globulin. AB - We have studied the binding of sex steroids to albumin and sex hormone binding globulin (SHBG) using gel filtration chromatography for the separation of the bound from the free fraction of the steroid. It was found that estradiol binds to the globulin and albumin in a nonlinear manner: a lag period of binding was observed at low concentrations of the proteins, followed by an exponential increase of the bound hormone as the protein concentration increased. The same was observed with dihydrotestosterone (DHT) and albumin but not with globulin. In the presence of a constant concentration of albumin, the increase of SHBG concentrations resulted in a rapid transfer of estradiol from albumin to globulin while the transfer of DHT was moderate. When whole serum was used, the increase of its amount again resulted in the transfer of estradiol from albumin to globulin. Our study showed that a substantial increase of globulin-bound hormone can occur, following small variations of the protein. This offers obvious advantages to the organism, by saving energy, material and time and plays a basic role in estradiol transfer from albumin to the much more biologically active globulin. PMID- 9358205 TI - Assessment of antibiotic levels in lung tissue with erosion-controlled dosage forms. AB - The concentration-time curve of ciprofloxacin has been assessed in the blood compartment and in the lung tissue following oral administration of the drug. Immediate release and erosion-controlled dosage forms have been examined. A numerical model based on finite differences and taking into account all relevant data has been built: the kinetics of drug release in the gastrointestinal tract, drug absorption in the blood compartment and elimination, and the transient diffusion of the drug through the lung tissue. A partition coefficient for the drug at the tissue-blood interface has been considered to express the drug concentration at the tissue surface. The effect of dose frequency and erosion rate on the antibiotic versus time curves in the plasma and the lung tissue has been studied in detail. PMID- 9358206 TI - The major metabolite of dipeptide piracetam analogue GVS-111 in rat brain and its similarity to endogenous neuropeptide cyclo-L-prolylglycine. AB - The metabolism of a new piracetam analogue, the dipeptide cognitive enhancer N phenylacetyl-L-prolylglycine ethyl ester (GVS-111) was studied in vivo. GVS-111 itself was not found in rat brain 1 h after 5 mg/kg i.p. administration up to limit of detection (LOD) under high performance liquid chromatography (HPLC) conditions. Three substances corresponding to the three possible GVS-111 metabolites, namely phenylacetic acid, prolylglycine and cyclo-prolylglycine, were found in experimental rat brain samples as well as in controls using HPLC, gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) methods. Only cyclo-prolylglycine concentration increased (2.5-fold) 1 h after GVS-111 administration. Cyclo-prolylglycine formation from GVS-111 in the presence of plasma and brain enzymes was shown in vitro. These data could be considered as evidence that GVS-111 is prodrug which converts in the body to cyclo prolylglycine, and which is identical to the endogenous cyclopeptide that produces the nootropic activity. PMID- 9358207 TI - Testosterone sulphation and glucuronidation in the human liver: interindividual variability. AB - Presystemic sulphation and glucuronidation at OH-C17 limits the bioavailability of testosterone; the aim of this investigation was to describe the variability in testosterone sulphation and glucuronidation rates in the human liver. Liver samples were obtained from 61 women and 40 men of similar age (mean 53 and 55 years, respectively) submitted to surgery. The mean rate of testosterone sulphation was significantly (P = 0.002) higher in men (22.4 pmol/min/mg) than in women (17.5 pmol/min/mg), was not age-dependent, followed bimodal distribution and varied over 7-fold in men and women. There was a weak, but significant negative correlation (r = -0.380; P = 0.003), between the rate of testosterone glucuronidation and age in the liver of women but not in that of men. The mean rate (pmol/min/mg) of testosterone glucuronidation was 155 (men) and 105 (women) (NS) and varied over 20-fold. When the rate of testosterone glucuronidation was expressed on the basis of g liver equivalent, the mean estimates were significantly (P = 0.003) greater in men (3323 pmol/min/g) than in women (1841 pmol/min/g). The present findings are consistent with the view that the hepatic activities of sulphotransferase and glucuronosyltransferase are higher in men than in women and that they vary in the human liver. PMID- 9358208 TI - Comparative pharmacokinetics of two nifedipine products in capsule form following single oral administration in healthy volunteers. AB - The pharmacokinetic parameters (AUC, Cmax, Tmax, and t1/2) of nifedipine following single oral administration of a 10 mg capsule of test product were compared to those of the same amount of a reference product. The two products in capsule form were administered according to a randomized two-way crossover design in 22 healthy male volunteers. Nifedipine plasma concentrations were determined using a rapid, sensitive and precise high performance liquid chromatography (HPLC) method with ultraviolet (UV) detection at 235 nm. The parametric 90% confidence intervals of the mean value of the ratio [Myogard (test product) /Adalat (reference product)] for pharmacokinetic parameters were 0.90-1.08, 0.80 1.07, and 0.93-1.12 for AUC0-->infinity, Cmax and t1/2, respectively. In each case, values were within the acceptable bioequivalence range of 0.8-1.25. Distribution free point estimate for the difference in expected medians of Tmax of the two products (Myogard-Adalat) was 0.00 h with a 90% confidence interval of 0.00-0.13 which is greater than the accepted bioequivalence of +/- 0.12. The kinetic parameters were comparable to those reported for nifedipine, and no statistically significant differences were found in any of them when comparing the two products by analysis of variance (ANOVA) on log-transformed data. Thus, the two products could be considered bioequivalent regarding absorption rate (Cmax and Tmax), extent of absorption (Cmax and AUC) and elimination (t1/2). PMID- 9358209 TI - Individualization of theophylline infusion rate on the basis of a nonlinear compartmental pharmacokinetic model. AB - In the present paper, a nonlinear compartmental model for theophylline pharmacokinetics is developed. The analytical solution of the model, in parametric form, is derived under plateau conditions for plasma metabolite concentration. The parameters are obtained from plasma and urine data using best fitting techniques and their values are used in order to calculate maintenance intravenous infusion. Numerical simulation is then performed in order to compare the drug concentration obtained by our approach with that of alternative intravenous regimens. The differences argue for individualized dosage regimens, since theophylline is a drug with a narrow therapeutic window and its concentration at the active sites strongly depends on characteristic parameters of the patient's response. Our results show that it is possible to estimate the patients' parameters during the first 8 h after intravenous administration of the drug and these parameters can be used to design an individualized dosage regimen in patients receiving theophylline intravenously. PMID- 9358210 TI - Management of bleeding, transfusion requirement and removal of catheters in transurethral prostate resection. AB - OBJECTIVE: The objective of the present study was to establish the therapeutic value of early coagulation of severe postoperative bleeding after transurethral prostate resection in unselected patients. PATIENTS AND METHODS: In a prospective study of 772 prostate resections carried out in 617 patients, bleeding complications, hemostatic measures, blood transfusions, catheter removals, and catheter-related and general complications were registered, and the factors influencing them were analyzed. RESULTS: Severe postoperative bleeding was coagulated endoscopically on the day of the operation in 70 resections (9.1%), and after removing the catheter in a further 19 cases (2.7%). Blood was transfused perioperatively in 14 patients (2.3%): in 11 patients (2.0%) because of preoperative anemia, and in 3 patients (0.3%) because of postoperative hemorrhage. In 96.3% of the resections, the catheter was removed on the first postoperative day, and in the last year of the study in 99.3% of the cases. Neither additional hemostasis nor early catheter removal had disadvantageous consequences. CONCLUSIONS: Transurethral prostate resections can be performed without any blood transfusion in more than 99.0% of patients without preexisting risk when severe postoperative hemorrhage is coagulated at an early stage. Moreover, this enables early catheter removal, after 24 h at the latest, in more than 99.0% of the cases. PMID- 9358211 TI - Reference ranges for the concentrations of total and complexed plasma prostate specific antigen and their ratio in patients with benign prostate hyperplasia. AB - OBJECTIVE: To establish the normal distribution and reference ranges of complexed and total prostate-specific antigen (PSA) and the complexed-to-total PSA ratio according to the age of the patients, so that PSA can be used to distinguish between prostate cancer and benign prostate hyperplasia (BPH). MATERIAL AND METHODS: Using specific ELISAs, total PSA and PSA complexed to alpha 1 antichymotrypsin (complexed PSA) were determined in 237 BPH patients, 160 with histologically confirmed BPH and 77 in whom prostate cancer was excluded by digital rectal examination, transrectal ultrasound and total PSA measurement. RESULTS: Both total and complexed PSA correlated with patient age (r = 0.424 and r = 0.379, p < 0.0001, respectively). However, no correlation was found between the complexed-to-total PSA ratio and age (r = 0.026, p > 0.2). The mean complexed to-total PSA ratio for the 237 BPH patients was 0.69 +/- 0.11, and only 23 had a ratio > 0.8. CONCLUSIONS: These results show that the cut-off point of 0.8 established for the complexed-to-total PSA ratio is the same for men of all ages, and that the use of this ratio may avoid many negative prostate biopsies, confirming that PSA: alpha 1ACT is a potential marker for differential diagnosis of prostate carcinoma and BPH. PMID- 9358212 TI - Clinical stage, prostate-specific antigen and Gleason grade to predict extracapsular disease or nodal metastasis in men with newly diagnosed, previously untreated prostate cancer. A multicenter study. A. Ur. O. Cooperative Group. AB - OBJECTIVE: To draw nomograms for preoperative predictions of extracapsular and of nodal disease based upon preoperative prostate-specific antigen, Gleason grade and clinical stage. METHODS: The complete charts of 1,738 patients submitted to radical retropubic prostatectomy in 34 Italian urological departments have been reviewed. The correlation between preoperative variables and pathological examination was tested by both univariate and multivariate techniques. Logistic regression analysis with the likelihood ratio chi 2 test was used to predict the pathological features (T > = 3; N+) of a patient for various combinations of preoperative variables. RESULTS: Probability plots were constructed for the prediction of either extracapsular disease or lymph node involvement by the above mentioned combination of preoperative variables. CONCLUSIONS: The obtained probability curves could be useful for patient counselling, for planning a staging laparoscopic lymphadenectomy in high-risk patients and for deciding whether to perform a nerve-sparing prostatectomy. PMID- 9358214 TI - Endoscopic correction of intractable stress incontinence with silicone micro implants. AB - OBJECTIVE: To assess the clinical value of silicone micro-implants (Macroplastique, Uroplastique, Bioplastique) in the treatment of intractable genuine stress incontinence. MATERIALS AND METHODS: Between January 1992 and March 1995, 34 patients with failed previous surgery for genuine stress incontinence were treated with transurethral submucosal injection of Macroplastique. RESULTS: The initial success rate was high, 90% at 1 month and 75% at 3 months postoperatively. However, at 2 years of follow-up the success rate fell to 48%. Patient satisfaction was high. CONCLUSION: Macroplastique is a relatively simple and effective day-case procedure for intractable genuine stress incontinence. It may have a place as a last-resort surgery for genuine stress incontinence predominantly due to sphincter incompetence in an unfit patient with failed previous anti-incontinence procedures despite good repositioning of the bladder neck. However, many more patients and a much longer follow-up are needed before any further meaningful statements can be made about the success or failure of this material. PMID- 9358215 TI - Urethroplasty by superficial membranous fascia for long urethral strictures: a new approach. AB - 36 patients has single-stage repair of severe bulbar urethral strictures using a superficial membranous fascia tubed flap. The length of follow-up varied from 9 months to 2 years (mean 15 months). Recurrence occurred in 1 case. Urodynamic studies in 35 cases before and after urethroplasty showed a marked improvement in urinary flow and voiding postoperatively. This procedure is safe, simple, economically preferable and has a no higher risk than other 1- and 2-stage procedures. PMID- 9358213 TI - 5-Fluorouracil versus folinic acid and 5-fluorouracil in advanced, hormone resistant prostate cancer: a prospective randomized pilot trial. AB - OBJECTIVES: Results of cytotoxic chemotherapy for hormone-resistant prostate cancer are not impressive. One of the substances which seems to have a therapeutic benefit is 5-fluorouracil (5-FU). The effect of 5-FU can be modulated by addition of folinic acid (FA). We tested in a prospective, randomized phase II trial monotherapy with 5-FU versus the combination of 5-FU and high-dose FA. METHODS: 25 patients received 600 mg/m2 5-FU, and 24 patients 400 mg/m2 FA plus 600 or 400 mg/m2 5-FU. They were treated for two cycles for 5 days in a 21-day interval followed by a weekly single-day application until progression occurred. Pain remission, toxicity, time to progression and survival were evaluated. RESULTS: Both regimens led to a pain remission in nearly 70% of the patients. Mucosal side effects like diarrhea and stomatitis occurred more often in the combination arm, whereas leukopenias were more frequent in the monotherapy are. We observed no statistically significant difference between the two treatment arms regarding time to progression and survival. CONCLUSIONS: Although both regimens led to a pain remission, side effects are too severe to recommend these protocols for standard treatment of hormone-resistant prostate cancer. PMID- 9358216 TI - Spot urine samples for the metabolic evaluation of urolithiasis patients. AB - OBJECTIVES: This prospective study was initiated to assess the significance of spot urine specimens (SU) for the metabolic evaluation of stone formers. METHODS: 68 stone patients (51 males, 17 females) and 20 controls (9 males, 11 females) participated. On 3 consecutive days, urine was collected. Fasting (SU1) and postprandial (SU2) SU were obtained. From these, aliquots were taken, all the other urine was mixed to obtain 24-hour timed specimens (24hU). In all specimens, pH, specific gravity, creatinine, calcium, magnesium, phosphate, citrate, uric and oxalic acids were measured. The latter analytes were related to creatinine (mmol/g creatinine). Pearson correlation coefficients with their levels of significance and the day-to-day variation were calculated. Using the values in the control group, normal values (means +/- 2 SD) were established. RESULTS: There was a significant correlation (p < 0.0001) between SU and 24hU for all parameters examined. The day-to-day variation of all analytes was considerable in SU and 24hU. CONCLUSIONS: Despite a minor inaccuracy by relating parameters to creatinine, SU are sufficient in the routine metabolic evaluation of stone formers, since a third of all 24hU has to be rejected because of considerable collection errors. SU circumvent this problem. Because of the day-to-day variation, 3 SU should be obtained to overcome the doubtful significance of one single specimen. PMID- 9358217 TI - Piezoelectric extracorporeal shock-wave lithotripsy of lower pole nephrolithiasis. AB - OBJECTIVE: To evaluate the efficacy of the EDAP LT 02 lithotriptor for the treatment of lower pole nephrolithiasis. METHODS: From January 1994 to September 1995, 91 patients presenting with solitary radiopaque calculi of the lower pole calix were treated by piezoelectric extracorporeal shock-wave lithotripsy (ESWL) with the EDAP LT 02. Among them, 82 were available for follow-up. The stones' largest diameter of these patients varied from 5 to 15 mm (mean = 8.1). Indications for ESWL were pain in 63 (77%), hematuria in 5 (6%), associated infection in 6 (7.5%) and stone size in 8 (10%) asymptomatic patients. Stone localization was assessed as very easy in 74 cases (90%) and difficult in 8 cases (10%) but no intraoperative IVP was needed. ESWL sessions were performed with intravenous sedo-analgesia in 69 cases (80%) and general anesthesia in 17 cases (20%). After ESWL we advised patients to combine a diuresis with postural drainage. RESULTS: Most patients were treated with one session of ESWL: none required more that two (mean = 1.05). The mean hospital stay for one session was 1.2 +/- 0.7 days. Obstructive complication rate was 11% and auxiliary treatments were necessary in 6%. The stone-free rate of in situ piezoelectric ESWL monotherapy was overall 84%, and 95% of patients with pain were cured. CONCLUSION: In the absence of abnormality of the upper urinary tract, the vast majority of small lower pole caliceal stones can be completely removed by piezoelectric ESWL without recourse to more invasive methods. PMID- 9358218 TI - Growing teratoma syndrome: experience of a single institution. AB - OBJECTIVE: To analyze the clinical outcome of patients diagnosed with growing teratoma syndrome (GTS) at a single center during a long follow-up. PATIENTS AND METHODS: Eleven patients with GTS are reported. GTS lesions were located in the metastatic sites involved at disease presentation. Involved sites were: retroperitoneum in 9 patients; lung in 3; supraclavicular lymph nodes in 2, and inguinal lymph nodes in 1. Surgical resection of the masses was the treatment of choice. RESULTS: Twenty-four surgical procedures were performed: 4 thoracotomies; 2 supraclavicular; 1 inguinal, and 17 retroperitoneal node resections. Three patients have not relapsed since surgery of the masses, at 37+, 110+ and 118+ months. Eight patients have relapsed, 6 with mature teratoma and 2 (22%) with cancer. To date, all the patients are alive, 6 of them without disease and 5 with teratoma after resection of the masses. CONCLUSIONS: GTS is an infrequent entity. Involved sites are only at locations previously affected by the disease. The treatment of choice is surgical resection but recurrence is common. Efforts should be done to complete resection of the masses. PMID- 9358219 TI - Correlation between semen parameters and retrograde flow into the pampiniform plexus before and after varicocelectomy. AB - OBJECTIVE: To compare, before and after repair of varicocele, the semen parameters (SP) in relation to the persistence of retrograde vein flow. MATERIAL AND METHODS: We examined the correlation between semen characteristics and retrograde flow (RF) into the pampiniform plexus in 158 patients with varicocele, before and after low inguinal spermatic vein ligation. RESULTS: The results of the investigation before surgery showed that all patients had RF and also a decrease in quantitative and qualitative SP. After surgery an improvement was observed in SP and there was a decrease in RF as a function of time elapsing from surgery. One year after surgery, an improvement in SP in 136 (86.1%) patients was observed. In 14 (8.8%) no change was noted and in 8 (5.1%) there was a deterioration in SP. Comparison between increased SP and persistence of RF 1 year after surgery revealed that of the 136 patients who showed improvement in SP, 124 (91.1%) did not have RF. Of the 14 patients with no improvement in SP, 12 (85.7%) were free of RF and 9 patients with deteriorated SP and bilateral RF. Amongst 132 infertile men, the wives of 55 (41.6%) became pregnant. None of these 55 men demonstrate RF. CONCLUSIONS: These results revealed a significant correlation between the improvement of SP, pregnancy in patients' wives and disappearance of RF. These results may be considered as a parameter of successful varicocelectomy. PMID- 9358221 TI - Endoscopic management of ureteroceles in children. AB - OBJECTIVE: Treatment of ureteroceles in children varies according to the anatomicopathological form and the choice of the surgical team. This study tries to determine the exact value of the endoscopic management of ureteroceles in children. METHODS: Between 1987 and 1993, 11 ureteroceles in 10 children were treated by endoscopic incision: 7 intravesical ureteroceles (4 single system and 3 duplex system) and 4 duplex-system ectopic ureteroceles. The procedure consists of a tiny transversal incision at the lower and median aspects of the ureterocele. RESULTS: The dilation of the upper urinary tract disappeared or decreased in all cases of intravesical ureteroceles and in half the cases of ectopic ureteroceles. Endoscopic incision of the ureterocele led to a vesicoureteral reflux in the associated ureter in 6 cases: 54.5% (43% of the intravesical ureteroceles, 75% of the ectopic ureteroceles). Following endoscopic treatment, no further surgery was required in 5 of the 7 cases with intravesical ureteroceles (71.5%), while every case of ectopic ureterocele needed a further operation (lower tract surgery in 3 cases, upper tract surgery in 1 case). CONCLUSIONS: Endoscopic incision of ureteroceles is a simple and quick procedure which allows obstruction to be removed and the dilation of the upper urinary tract and its corresponding kidney function to be improved, particularly in the neonate. The endoscopic management of ureteroceles may in itself suffice, without necessity of further surgery. These favorable results can more readily be seen in cases of intravesical ureterocele than in those of ectopic ureterocele. PMID- 9358220 TI - Urinary incontinence in Belgium: a population-based epidemiological survey. AB - OBJECTIVE: To investigate the prevalence, typology, and experience of urinary incontinence, as well as the available therapeutic modalities and information sources among people living at home in Belgium. METHODS: A representative population sample of 5,269 adults completed a questionnaire in their own homes. There were 2,499 men and 2,770 women aged 30 years and over. RESULTS: 130 men (5.2%) and 442 women (16.30%) had urinary incontinence at the time of the survey. Of 1,426 women aged over 50 years, 300 (21.0%) had urinary incontinence. 17.4% of the incontinent people experience several episodes daily. In women, incontinence increases with parity. Stress incontinence is by far the most common form of the disease (42%); 53% of incontinent women (9.9% of the female population) experience this kind of disorder. Overall, 95% of stress incontinent people are women. The relative frequency of urge incontinence is similar in both sexes (males 45%; females 55%). Urinary incontinence is considered as bothersome by about 30% of the affected subjects; 7 of 10 subjects with daily incontinence episodes consider the disorder as more or less bothersome. 29.9% of affected people report that they discuss the problem with their general practitioner, 11.4% with a specialist, 25.3% with their spouse or partner, and 17.4% with a family member. Of course, people who consider the disease as bothersome score higher on these items. On the other hand, 32.1% do not reveal the problem. Although 72.5% of incontinent people are aware of protection systems, the main therapeutic modalities are not very well known (drugs 33.9%; surgery 24.7%). Most people with incontinence (70-75%) take no specific measures, 20% use pads, panty liners or nappies, and only 9% look for a curative measure. The preferred information source is the family physician for 50%, the specialist physician for 11%, and the pharmacist for 9%. The role of the mass media is equivalent to that of specialist physicians and pharmacists. CONCLUSIONS: Incontinence has a profound effect on daily life, and is still considered by many as 'taboo'. An appropriate information system, in which the general practitioner plays a key role, is obviously desirable. PMID- 9358222 TI - Unicalyceal kidney associated with ureteral anomalies. AB - A unicalyceal kidney is a very rare anomaly of the urinary system. We report 4 cases of unicalyceal kidneys in infants. All cases were associated with ureteral anomalies: megaureter on the involved side, and/or ectopic ureter with vesicoureteral reflux or renal agenesis on the contralateral side. In order to preserve renal function we recommend surgical treatment for patients with unicalyceal kidney when they have ureteral anomalies. PMID- 9358223 TI - Changes in keratin expression during the development of benign prostatic hyperplasia. AB - OBJECTIVE: The relationship between different types of epithelial cells in the prostate and the regulatory mechanism underlying benign prostatic hyperplasia (BPH) are still obscure as is the association between BPH and prostate carcinoma (PCa.) On the basis of keratin immunophenotyping, a subpopulation of cells in normal rat prostate and human PCa have been identified as candidates for the 'amplifying cell' in the stem cell model. In this model the basal cell is described as being associated with the stem cell. From this precursor an intermediate cell type develops which may differentiate into the luminal-type cell. In this study these different cell types are investigated in the development of isolated BPH and BPH associated with PCa, using monoclonal antibodies to intermediate filaments of the keratin class. METHODS: We immunohistochemically stained 64 snap-frozen human prostatic tissues, using monoclonal antibodies against keratin 14 (marker for putative 'stem cell'), keratin 18 (marker for putative 'transit cell'), and MAb RCK103 (marker for putative 'amplifying cell' or intermediate cell). RESULTS: In basal cell hyperplasia, an atypical form of BPH, keratins previously associated with intermediate cells were frequently detected. Cells with this keratin phenotype were detected in the luminal compartment of BPH, and were more prevalent in BPH adjacent to PCa. This keratin expression pattern was similar to that of PCa. CONCLUSION: On the basis of keratin phenotyping we demonstrated that large numbers of cells with the keratin expression pattern of so-called intermediate cells were identified in BPH associated with PCa, while in isolated BPH these cells were infrequently found. This supports the concept that BPH with intermediate phenotype may have premalignant potential. Furthermore this is suggestive of an etiologic relationship between the two diseases. PMID- 9358224 TI - Photodynamic effects of sulfonated aluminum chlorophthalocyanine in human urinary bladder carcinoma cells in vitro. AB - OBJECTIVES: Topically applied photosensitizers are particularly suited for use in the photodynamic therapy of urinary bladder tumors. The objective of the present study was to investigate the photodynamic effects of sulfonated aluminum chlorophthalocyanine on human bladder carcinoma cells. METHODS: The photodynamic efficiency and the morphologic changes induced by sulfonated aluminum chlorophthalocyanine on human bladder carcinoma cells and normal bladder wall cells were investigated in vitro using light microscopy and in order to determine the subcellular target organelles using electron microscopy. RESULTS: Examination of tumor cells with light microscopy after previous photodynamic therapy initially showed distension of the cells and, depending on the duration, a significant blistering of the cell membrane, leading, in some cases, to rupture of the cells. Electron microscopy revealed destruction of the mitochondria. No changes in other cell organelles or structures were observed. CONCLUSIONS: Sulfonated aluminum chlorophthalocyanine exhibits good photodynamic activity in bladder tumor cells with relative tumor selectivity. The primary cellular target of photodynamic therapy seems to be the mitochondria. PMID- 9358225 TI - Artificial stimulation of cryopreserved human spermatozoa by sodium nitroprusside, 2-chloroadenosine, and 2-deoxyadenosine. AB - OBJECTIVE: We analyzed the effects of sodium nitroprusside (SNP), 2 chloroadenosine (2-CLA), and 2-deoxyadenosine (2-DA) on sperm motility and motion characteristics of cryopreserved human spermatozoa. METHODS: Thawed semen samples from healthy donors (n = 10) were incubated with the stimulants for 0, 30, 60, 120, and 180 min. The final concentrations used were SNP 25, 50, and 100 nM; 2 CLA 12.5, 25, and 50 microM, and 2-DA 0.5, 1, and 2.5 mM. Sperm motility and changes in motion characteristics were measured on a computer-assisted semen analyzer. RESULTS: Compared with control (0 min, no stimulant), a significant improvement in percentage motility was generally seen with all three stimulants even at 180 min. The sperm motility improved at concentrations of 50 and 100 nM for SNP (except at 120 min), 25 and 50 microM for 2-CLA, and at all concentrations of 2-DA. Other sperm motion characteristics increased to varying extent with the stimulants. Of the three stimulants, all concentrations of 2-DA resulted in significant increases in curvilinear velocity, average path velocity, and amplitude of lateral head displacement at 60 min of incubation. CONCLUSIONS: All three stimulants significantly increased percentage motility for extended intervals of time. 2-DA also improved sperm motion characteristics, though these changes were less uniform with 2-CLA. Motility stimulation by these chemicals may be beneficial in the treatment of cryopreserved and oligozoospermic sperm specimens for use in assisted reproduction. PMID- 9358226 TI - Comparison of muscle histology and generated pressures of two types of dynamic graciloplasties in rabbits. AB - OBJECTIVE: To compare the muscle histology and the generated pressures of the conventional spiral graciloplasty with those of the split sling graciloplasty in rabbits. METHODS: Six rabbits underwent a split sling graciloplasty in the left leg and a conventional graciloplasty in the right leg around polyurethane tubes. Beforehand muscle biopsy specimens were taken at several locations in both legs. After chronic stimulation once again biopsies were performed. Comparisons were made with regard to histology and pressures. RESULTS: The same level of global changes occurred in both legs. The type II fiber diameter increased significantly in both legs. The amount of connective tissue increased significantly in both legs, but the resulting percentages were comparable. The changes at the distal site of the split sling graciloplasty were comparable to other biopsy specimens. The mean pressures in the conventional graciloplasty were 42 and 52 cm H2O without and with stimulation, respectively. In the split sling graciloplasty these pressures were 48 and 76 cm H2O, respectively. The ability to sustain long lasting contractions was the same using both techniques. 25 Hz was the optimal frequency for muscle stimulation. CONCLUSIONS: Histologically the conventional graciloplasty is comparable with a split sling graciloplasty. The achieved pressures are the same or slightly higher with the split sling graciloplasty as compared with the conventional graciloplasty. PMID- 9358227 TI - Ultrastructural osteopontin localization in papillary stones induced in rats. AB - OBJECTIVES: To detect in situ the precise osteopontin (OPN) localization in papillary stones. METHODS: Immunocytochemical labelling procedures are applied to detect OPN localizations in crystalline material of renal papillary stones. The tissue-processing procedure for electron microscopy, which includes OsO4 postfixation, preserves both immunocytochemical OPN reactivity and cellular membrane contrast up to the ultrathin section. Reflection-contrast light microscopical images are correlated with high resolution transmission-electron microscopical observations from consecutive ultrathin epon sections. RESULTS: Preserved crystalline material in interstitial and peripheral papillary stones is recognized as calcium oxalate monohydrate. After section incubation with markers conjugated to an antibody against OPN (alpha OPN) the crystals are converted into ghosts. In the ghosts, alpha OPN markers are present around microcrystals. The size of these microcrystals ranges from several nanometers to micrometers. It is observed (due to the OsO4-preserved membranes) that interstitial cells are separated from the stone surfaces by unidentified extracellular material, also present in the center as a stone matrix. CONCLUSION: The microcrystal-growth inhibitor OPN is detected in situ in interstitial stones induced in the rat's papilla and at the surface of the papilla. PMID- 9358228 TI - Clinically occult Leydig cell tumor in a cryptorchid man. Report of a case presenting with unilateral gynecomastia and impotence. AB - Left painful breast enlargement and impotence were the main complaints of a right cryptorchid young man. On ultrasonography a hypoechoic mass was found in the right testis. Postoperatively the increased level of estrogens and the decreased levels of testosterone were normalized 6 months following the orchiectomy, gynecomastia subsided considerably and the impotence improved quite satisfactorily. PMID- 9358229 TI - Anterior mediastinal metastasis of testicular germ cell tumor: relation to benign thymic hyperplasia. AB - OBJECTIVE: To assess the anterior mediastinal mass in recurrent testicular cancer, with relation to thymic hyperplasia after treatment. METHODS: The anterior mediastinal regions were fully evaluated by chest computed tomography (CT) at the initial staging and after treatment in 24 of 44 patients with testicular cancer. RESULTS: One patient with stage IIB tumor had thymic hyperplasia before treatment, and one with stage III had benign thymic hyperplasia after chemotherapy with salvage surgery. Three of 4 patients who had recurrence had an anterior mediastinal mass. One had benign thymic hyperplasia confirmed by histology and 2 had metastatic tumor confirmed by histology and clinical course, in which the mass became so enlarged that it obstructed major vessels. CONCLUSION: Although the relationship of the CT finding to the response to treatment in the anterior mediastinal mass and other metastatic lesions provide some clues helpful in differentiating benign from malignant masses, surgical exploration is recommended for the patient with an indication for salvage surgery. PMID- 9358230 TI - Focal xanthogranulomatous pyelonephritis: partial nephrectomy as definitive treatment. AB - OBJECTIVES: We report 11 cases of focal xanthogranulomatous pyelonephritis (FXGP), a disease that is very uncommon. The aim of the present work is to assess the effectivity of conservative treatment. MATERIAL AND METHODS: Eleven of the 82 cases of XGP (12.5%) diagnosed between 1970 and 1995 presented the focal form (FXGF). Clinical features, laboratory findings, radiological imaging studies, surgical treatment and follow-up were evaluated. RESULTS: FXGP occurred in middle aged women (female/male ratio 4.5:1) who had a history of calculosis, urinary infections produced by Escherichia coli and Proteus mirabilis, or urinary tract abnormalities. FXGP was unilateral in all cases. The most frequent symptom was flank pain. Some hematological and biochemical parameters were altered. Intravenous urography and sonography revealed calculosis, hydronephrosis or renal mass, but these findings are nonspecific. Only abdominal CT scan can establish the correct diagnosis. Five of these patients (45.5%) underwent partial nephrectomy at our hospital. During the follow-up, patients showed no relapse in the ipsilateral or contralateral kidney. Serum parameters were in normal range. CONCLUSION: When FXGP is diagnosed, local excision is recommended in all cases, since relapse in the affected kidney is unusual. PMID- 9358231 TI - Chronic granulomatous disease masquerading as a bladder tumor: a potential source of diagnostic error. AB - Chronic granulomatous disease (CGD) is a rare inherited disease of childhood, characterized by recurrent bacterial or fungal infections. The underlying defect is a dysfunction of neutrophil granulocytes interfering with their ability to kill phagocytosed microorganisms. Genitourinary tract involvement has been reported in 38% of these patients. We report a case of CGD in whom the most important findings were urinary bladder tumors at different locations and a subsequent obstruction of the left ureter. A review of the pathogenesis of the disease, potential involvement of the urinary tract and treatment is presented. PMID- 9358232 TI - Protein kinase and phosphatase activity in the lungs of normoxic versus hyperoxic rats. AB - Studies were designed to examine aspects of phosphorylation and dephosphorylation in rat lung cells in response to hyperoxic exposure. Protein kinase and phosphatase activities were measured in preparations of lungs from normoxic rats, hyperoxia-exposed rats (95% O2 for 60h), and rats recovering in room air for 1 and 3 days. Protein kinase C (PKC) activity immediately postexposure was significantly lower than in normoxic controls (normoxia 127.1 +/- 13 vs. hyperoxia 101.5 +/- 6 pmol/min mg-1) and continued to decline during the recovery period (85.3 +/- 4 and 78.2 +/- 6 pmol/min mg-1 at 1 and 3 days recovery, respectively). The PKC activity did not translocate from cytoplasm to the membranes. In contrast, PKA activity did not change in response to hyperoxia exposure or recovery. Protein phosphatase activity was decreased significantly by hyperoxia exposure (normoxia 30.7 +/- 3 vs. hyperoxia 21.9 +/- 1 pmol/min microgram-1) but returned to normoxic control levels by 1 and 3 days (24.1 +/- and 31.5 +/- 1 pmol/min microgram -1, respectively). Protein phosphatase activity was inhibited by okadaic acid (Ki = 1 nM) and calyculin A (Ki = 0.61 pM), indicating a type 2A protein phosphatase. Enzyme activities in cultured type II alveolar cells paralleled those observed in whole lung preparations. Decreased enzyme activities in the lung may be located to the development of acute lung injury during hyperoxic exposure. PMID- 9358233 TI - Clearance of particles from small ciliated airways. AB - In recent years, there has been a debate on whether a considerable fraction of particles is retained after 24 h in the tracheobronchial region. In the present study, 8 healthy subjects inhaled 6.2-microns monodisperse Teflon particles labeled with 111 In twice, at flow rates of 0.45 and 0.045 L/s. According to theoretical calculations, the particles inhaled at 0.45 L/s should deposit mainly in large bronchi and in the alveolar region, whereas the particles inhaled at 0.045 L/s should be deposited mainly in small ciliated airways. Twenty-four hours after inhalation, about half of the particles inhaled with both modes of inhalation had cleared. Clearance during the period from 1 to about 30 days after inhalation, could, for both modes of inhalation, be described by the sum of two exponential functions. For the inhalation rate of 0.45 L/s, 15% cleared with a half-time of 3.4 days and 85% with a half-time of 190 days. For the inhalation of 0.045 L/s, 20% cleared with a half-time of 2.0 days and 80% with a half-time of 50 days. The results strongly indicate (1) that a considerable fraction of particles deposited in small ciliated airways had not cleared within 24 h, and (2) that these particles cleared differently from particles deposited in the alveolar region. The experimental data agree quite well with the IRTM predictions made using its default slow clearance fractions. PMID- 9358234 TI - Human bronchiolar deposition and retention of 6-, 8- and 10-micrograms particles. AB - Three groups, each consisting of 6 healthy subjects, inhaled, respectively, 6 micrograms (aerodynamic diameter), 8-micrograms, and 10-micrograms Teflon particles, labeled with indium-111. The particles were inhaled at an extremely low flow rate, 0.05 L/s. Lung retention was measured after 0, 24, 48, and 72 h. Two models were used to calculate particle deposition in the lungs in the various generations: the Karolinska Institute model (KI model) and the University of Southampton model (US model). From the experimental clearance data and the theoretical deposition data, it was calculated that the average retention after 24 h was around 100% for particles deposited in generations 13-16 (ciliated bronchioles) and around 20% in generations 0-12 (both large and small ciliated airways). In these calculations, it was assumed that the retained fractions were independent of particle size. The depositions in the bronchial region (generations 0-8), bronchiolar region (generations 9-15 or 9-16), and the alveolar region were calculated using the two models and compared with the recent ICRP model. On the whole, the three models agreed fairly well. PMID- 9358235 TI - Comparison of adult and newborn pulmonary cytokine mRNA expression after hyperoxia. AB - Neonatal animals of several species are more tolerant of hyperoxic exposure than are adults. However, the mechanisms of increased neonatal tolerance are unknown, as are the cell types that contribute to oxygen resistance. This study examined hyperoxic lung injury in neonatal and adult C57BL/6 mice. Adults and neonatal mice were exposed to > 95% oxygen for 78 h and 10 days, respectively. Lung mRNAs were assayed by RNase protection assay. After 72 h of exposure, the messages encoding tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta and 6 (IL 1 beta, IL-6) were increased 2-fold in adult lungs. However, at this time point these mice are near or at lethality. No alterations in neonatal lung mRNAs were detected until 7 days of oxygen exposure. At that time neonatal mice demonstrated increases in lung mRNAs encoding TNF-alpha, IL-1 beta, and IL-6 of 3-, 5-, and 8 fold, respectively. Acute alveolitis and slight edema were detected, but lethality wasn't observed until 10 days of exposure. In situ hybridization in neonatal mice suggests accumulation of TNF-alpha and IL-1 beta transcripts in pulmonary interstitial macrophages and in a subset of neutrophils after 7 days of exposure. Messages encoding IL-1 alpha, IL-2, IL-3, IL-4, IL-5,IL-10 interferon gamma (IFN-gamma), and TNF-beta were not altered from controls in either adult or neonatal mice at any time point examined. In conclusion, adult mice demonstrate little change in cytokine mRNA until lethality is imminent, whereas newborn mice demonstrate an acute induction of TNF-alpha, IL-1 beta, and IL-6 early in the development of hyperoxic injury, which suggests that a rapid cytokine response early in the development of hyperoxic injury may play an important role in the adaptation of neonatal lungs to toxicity from prolonged oxygen exposure. PMID- 9358236 TI - Hemodynamic stress enhances neutrophil responsiveness to chemotactic stimuli. AB - Circulating neutrophils are exposed to widely varying levels of hemodynamic stress induced by blood flow conditions. This study examined the effect of hemodynamic stress on the functional responsiveness of neutrophils obtained from healthy humans to chemotactic stimuli. To expose neutrophils to hemodynamic stress in vitro, isolated neutrophils were agitated under artificial flow conditions induced by a rotary tube apparatus. Although such hemodynamic stress produced no spontaneous or random migration of neutrophils, it enhanced neutrophil migration in response to the chemotactic peptide f-methionyl-leucyl phenylalanine (FMLP) by as much as 200%. Hemodynamic stress also enhanced polarization in response to FMLP, producing a change in shape characteristic of migration. Polarization was reversible when neutrophils were transferred to quiescent conditions after being exposed to hemodynamic stress. Hemodynamic stress also enhanced O2.- production and granular beta-glucuronidase release in response to FMLP and enhanced polarization and O2.- production in response to phorbol myristate acetate (PMA), a direct activator of protein kinase C. Extracellular Ca2+ was not required for the enhancement of chemotactic responsiveness by hemodynamic stress, and the stress produced no detectable change in intracellular Ca2+, intracellular cyclic AMP, or activated protein kinase C levels in neutrophils. The results show that hemodynamic stress enhances the functional responsiveness of neutrophils to chemotactic stimuli and provide insights into interpretation of in vitro data usually obtained from quiescent conditions. PMID- 9358238 TI - Cytochrome P-450 2E1 in rat liver peroxisomes: downregulation by ischemia/reperfusion-induced oxidative stress. AB - Cytochrome P-450 containing enzymes, known to be present in the endoplasmic reticulum and mitochondria, catalyze the oxidation of various compounds. In this study we have used highly purified peroxisomes (> 95%) to provide evidence by analytical cell fractionation, enzyme activity, Western blot, and immunocytochemical analysis that cytochrome P-450 2E1 (Cyp 2E1) is present in peroxisomes. Similar specific activities of aniline hydroxylase, a Cyp 2E1 dependent enzyme, in purified peroxisomes (0.72 +/- 0.03 nmol/min/mg protein) and microsomes (0.58 +/- 0.03 nmol/min/mg protein) supports the conclusion that peroxisomes contain significant amount of Cyp 2E1. This peroxisomal Cyp 2E1 was also induced in acetone-treated rat liver. The status of microsomal and peroxisomal Cyp 2E1 was also examined following ischemia/reperfusion-induced oxidative stress. Ischemia alone had no effect; however, reperfusion following ischemia resulted in decrease in Cyp 2E1 both in microsomes and peroxisomes. This demonstration of cytochrome P-450 2E1 in peroxisomes and its downregulation during ischemia/reperfusion describes a new role for this organelle in cytochrome P-450 related cellular metabolism and in oxidative stress induced disease conditions. PMID- 9358237 TI - Effects of retinol deficiency and hyperoxia on collagen gene expression in rat lung. AB - Exposure to hyperoxia results in lung injury and a decrease in lung collagen. Retinol is known to influence collagen gene expression, and retinol deficiency has been shown to potentiate hyperoxic lung injury. To investigate the combined effects of retinol deficiency and hyperoxia on lung collagen expression, retinol deficient rats were exposed to acute hyperoxia, and expression of the alpha-1 chains of type I procollagen [pro alpha 1 (I)] and type III procollagen [pro alpha 1 (III)] were determined using Northern hybridization analyses and immunohistochemical staining. Hyperoxia alone reduced pro alpha 1 (I) mRNA by 60 +/- 4% (p < .05) and pro alpha 1 (III) mRNA by 30 +/- 5% (p < .05), and retinol deficiency alone reduced pro alpha 1 (I) mRNA abundance by 49 +/- 8.8% (p < .05) and pro alpha 1 (III) mRNA abundance by 14 +/- 7.5% (p = not significant), respectively. Retinol deficiency plus hyperoxia did not cause any further reduction in procollagen mRNA than that seen with oxygen exposure alone. Immunohistochemical staining demonstrated decreased staining for type I collagen in retinol-deficient animals. Hyperoxic exposure resulted in decreased connective tissue staining and increased alveolar wall staining for type I collagen. Retinol deficiency and hyperoxia together resulted in a marked increase in alveolar exudates staining for type I collagen. No changes in type III collagen staining were seen. These findings demonstrate that while retinol deficiency does not potentiate hyperoxia-induced reductions in procollagen mRNA, it is associated with alterations in collagen staining in distal lung and immunohistologic evidence of collagen fragments in alveolar exudates. PMID- 9358239 TI - Gamma-irradiation damage to liposomes differing in composition and their protection by nitroxides. AB - The present study aims to determine the effect of bilayer composition on oxidative damage and the protection against it in lipid multicomponent membranes. Irradiation damage in 200-nm liposomes and the protection provided by the nitroxide radicals, 2,2,6,6-tetramethylpiperidine-1-oxyl (Tempo) and 4-hydroxy 2,2,6,6-tetramethylpiperidine--1-oxyl (Tempol) were assessed by monitoring several chemical and physical parameters. Liposomes were prepared in four different lipid compositions (mole ratios), DPPC:DPPG 10:1; DPPC:DPPG:cholesterol 10:1:4; EPC:EPG 10:1; and EPC:EPG:cholesterol 10:1:4, and gamma-irradiated with a dose of 32 kGy. Lipid degradation was determined by HPLC and GC analyses, whereas size and differential scanning calorimetry measurements were used to monitor physical changes in the liposomal dispersions. The results indicate that: (1) addition of 5 mM Tempo or Tempol, or freezing of the sample inhibited radiation induced lipid degradation; (2) Tempo and Tempol caused neither physical nor chemical changes in the liposomal dispersions; and (3) both nitroxides prevented or reduced some of the radiation-induced changes in thermotropic characteristics of the liposomes, preventing a shift in the temperature of the maximum of the main phase transition. PMID- 9358240 TI - Identification of possible reactive oxygen species involved in ultraviolet radiation-induced oxidative DNA damage. AB - We have previously demonstrated that each region of the ultraviolet (UV) spectrum (UVA, UVB, and UVC) induces the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in purified calf thymus DNA and HeLa cells in a fluence-dependent manner. In the present study, we further characterize the possible reactive oxygen species (ROS) that are involved in the induction of 8-oxodGuo by UV radiation. Sodium azide, a singlet oxygen (1O2) scavenger though its quenching effect on HO. was also reported, inhibited 8-oxodGuo production in calf thymus DNA exposed to UVA, UVB, or UVC in a concentration-dependent fashion with maximal quenching effect of over 90% at a concentration of 10 mM. Catalase, at a concentration of 50 U/ml, reduced the yields of UVA- and UVB-induced 8-oxodGuo formation by approximately 50%, but had little effect on UVC-induced 8-oxodGuo production. In contrast, 50 U/ml of superoxide dismutase (SOD) did not affect induction of 8-oxodGuo by any portion of the UV spectrum. Hydroxyl radical (HO.) scavengers mannitol and dimethylsulfoxide (DMSO) moderately reduced the levels of 8-oxodGuo induced by UVA and UVB, but not those by UVC. Instead, mannitol and DMSO enhanced the formation of 8-oxodGuo induced by UVC. These results suggest that certain types of ROS are involved in UV-induced 8-oxodGuo formation with 1O2 playing the predominant role throughout the UV spectrum. Except for UVC, other ROS such as hydrogen peroxide (H2O2) and HO. may also be involved in UVA- and UVB induced oxidative DNA damage. Superoxide anion appears not to participate in UV induced oxidation of guanosine in calf thymus DNA, as SOD did not display any quenching effects. PMID- 9358241 TI - Hydroxyl radical formation in hyperglycemic rats during middle cerebral artery occlusion/reperfusion. AB - Preexisting hyperglycemia is associated with enhanced reperfusion injury in the postischemic rat brain. The goal of this study was to evaluate whether the hyperglycemic exacerbation of brain injury is associated with enhanced generation of hydroxyl radicals in rats subjected to middle cerebral artery occlusion (2 h), followed by reperfusion (2 h). Magnetic resonance images revealed the exacerbation of focal brain injury in hyperglycemic rats. The salicylate trapping method was used in conjunction with microdialysis to continuously estimate hydroxyl radical production by measurement of the stable adducts 2,3- and 2,5 dihydroxybenzoic acid (DHBA) during ischemia/reperfusion. In normoglycemic rats, from a mean baseline level of 130 nmol/l, 2,3-DHBA levels surged to peak levels of 194 nmol/l 45 min into ischemia and to 197 nmol/l 15-30 min into the reperfusion period, returning to baseline by 2 h into reperfusion. A similar temporal profile was observed in hyperglycemic rats, except that absolute 2,3 DHBA levels were higher (165 nmol/l at baseline, 317 nmol/l peak during ischemia, 333 nmol/l peak during reperfusion), and levels remained significantly high (p < .05) throughout the reperfusion period. These results suggest that hydroxyl radical is an important contributor to the exacerbation of neuronal and cerebrovascular injury after focal ischemia/reperfusion in hyperglycemic rats. PMID- 9358242 TI - Reactive oxygen species contribute to epidermal hyaluronan catabolism in human skin organ culture. AB - Hyaluronan (HA) is produced by keratinocytes in human skin organ culture, and degraded locally in epidermis by an unknown metabolic route. The present work tested whether reactive oxygen species (ROS), spontaneously produced in the tissue, could contribute to HA catabolism in epidermis. Epidermal HA was endogenously labeled with 3H-glucosamine for 24 h, then chased for 24 h in the presence of superoxide dismutase (SOD) and catalase to reduce the concentration of ROS. In control cultures, 35% of labeled HA was degraded during the 24 h chase while the corresponding figures in the presence of SOD and catalase were 19% and 23%, respectively (p < 0.05). Methionine, a quencher of hypochlorous acid, did not significantly inhibit the degradation. In additional experiments, the iron and copper chelator Detapac was even more effective, reducing the degradation to 8-9%, and suggesting that the ROS responsible for the degradation were produced in the Fenton reaction. Dermal HA, and proteoglycans in both epidermis and dermis were not influenced by the treatments, indicating that the inhibition by SOD, catalase and Detapac on epidermal HA catabolism was specific. It is suggested that endogenous ROS is involved in the catabolism human epidermal HA. PMID- 9358243 TI - Antioxidant properties of S-adenosyl-L-methionine: a proposed addition to organ storage fluids. AB - Glutathione (GSH) depletion adversely affects the survival of organ grafts. Supplementation of commercial organ preservation solutions with GSH is complicated by the ease of oxidation of its thiol group and its ability to act as a pro-oxidant under certain conditions. Alternative sulphur-containing compounds such as S-adenosyl-L-methionine (SAM) can reduce ischaemia-reperfusion injury, possibly by acting as glutathione precursors, and are effective when added to preservation solutions. Although the antioxidant properties of GSH are known in some detail, there is little information on the ability of SAM to interact directly with reactive oxygen species (ROS) produced during ischaemia-reperfusion injury. This work compares the interaction of SAM and GSH with several ROS which may be formed during ischaemia-reperfusion. In a variety of lipid peroxidation systems, SAM and GSH had little effect except at high concentrations (5 mM) where they became pro-oxidant. Scavenging of O2.- by both species was slow. SAM was less effective than GSH at preventing damage by peroxynitrite or HOCl. In contrast, SAM was more effective than GSH in scavenging hydroxyl radicals (.OH) and in chelating iron ions to inhibit .OH generation. Unlike GSH, SAM did not stimulate .OH formation at low concentrations. The beneficial effects of SAM in preservation solutions could therefore include direct radical scavenging as well as acting as a precursor for intracellular GSH. PMID- 9358245 TI - Free radical-induced tandem base damage in DNA oligomers. AB - A new tandem base lesion has been identified in two DNA oligomers, namely d(GpT) and d(CpGpTpA), exposed to X-irradiation in deoxygenated aqueous solution. In this lesion the C6 carbon atom of thymine is hydroxylated and a covalent link is formed between the C5 carbon atom of thymine and the C8 carbon atom of the adjacent guanine base. In addition, further evidence in the form of mass spectrometric data is presented confirming the structures of previously reported tandem base lesions that are produced by ionizing radiation in the presence of oxygen. New data is presented on the prevalence of a previously reported tandem base lesion in which the methyl carbon atom of thymine is covalently linked to the C8 carbon atom of the adjacent guanine base. The free radical-initiated processes by which tandem base damages are generated are discussed. To date four different radiation-induced tandem base lesion have been identified. The evidence suggests that tandem base damage is a significant component of free radical induced DNA damage. PMID- 9358244 TI - Benzophenone-sensitized photooxidation of sarcoplasmic reticulum membranes: site specific modification of the Ca(2+)-ATPase. AB - Benzophenone (BP) was used as a photosensitizer to initiate lipid peroxidation in model and native biological membranes at concentrations of BP that do not perturb bilayer structure, as assessed by stearic acid spin label dynamics. Illumination of BP partitioned into sarcoplasmic reticulum membranes (SR) results in an exponential decay of BP and a linear accumulation of conjugated dienes and other products of lipid peroxidation as observed previously for micelles of linoleic acid [Marcovic and Patterson. Photochem. Photobiol. 58:329-334, 1993]. Lipid peroxidation was substantially inhibited in the presence of membrane-spanning proteins in SR compared to protein-free lipid vesicles, suggesting the competitive reaction of the initiator (triplet BP) and BP-derived radical species with protein groups. Modification of the predominant integral membrane protein, the Ca(2+)-ATPase, was demonstrated by changes in Ca(2+)-ATPase amino acid composition as well as by its functional inhibition. The rate of calcium transport showed an immediate exponential decay to completion, while calcium dependent ATPase activity exhibited an initial lag before modest inactivation. These results are consistent with the respective localization of calcium transport sites within membrane-spanning peptides and the ATP-binding site within the cytosolic domain of the Ca(2+)-ATPase, further suggesting that photosensitization of BP models oxidative stress inside the hydrophobic interior of the SR membrane. PMID- 9358246 TI - alpha-Phenyl-N-tert-butylnitrone attenuates excitotoxicity in rat striatum by preventing hydroxyl radical accumulation. AB - Various in vitro experiments have indicated that oxygen-derived free radicals may contribute to excitotoxic neuronal death. In the present study we induced excitotoxicity in rat striatum by perfusing glutamate at a high concentration through a microdialysis probe. We observed an increased formation of hydroxyl radicals (.OH) during the perfusion of the excitotoxin and an extensive striatal lesion 24 h after the insult. The spin trap, alpha-phenyl-N-tert-butylnitrone (PBN), attenuated both hydroxyl radical levels and the volume of the lesion. This result suggests that the neuroprotection may be due to a free radical scavenging mechanism. It also implies that PBN may be used in pathological situations involving excitotoxicity such as stroke, brain trauma, and chronic neurologic diseases. PMID- 9358247 TI - Examination of the inhibitory effect of norathyriol in formylmethionyl-leucyl phenylalanine-induced respiratory burst in rat neutrophils. AB - Norathyriol, aglycone of a xanthone C-glycoside mangiferin isolated from Tripterospermum lanceolatum, concentration dependently inhibited the formylmethionyl-leucyl-phenylalanine (fMLP)-induced superoxide anion (O2.-) generation and O2 consumption in rat neutrophils. In cell-free oxygen radical generating system, norathyriol inhibited the O2.- generation during dihydroxyfumaric acid (DHF) autoxidation and in hypoxanthine-xanthine oxidase system. fMLP-induced transient elevation of [Ca2/]i and the formation of inositol trisphosphate (IP3) were significantly inhibited by norathyriol (30 microM) (about 30 and 46% inhibition, respectively). Norathyriol concentration dependently suppressed the neutrophil cytosolic phospholipase C (PLC). In contrast with the marked attenuation of fMLP-induced protein tyrosine phosphorylation (about 70% inhibition at 10 microM norathyriol), norathyriol only slightly modulated the phospholipase D (PLD) activity as determined by the formation of phosphatidic acid (PA) and, in the presence of ethanol, phosphatidylethanol (PEt). Norathyriol did not modulate the intracellular cyclic AMP level. In the presence of NADPH, the phorbol 12-myristate 13-acetate (PMA) activated particulate NADPH oxidase activity was suppressed by norathyriol in a concentration-dependent manner and the inhibition was noncompetitive with respect to NADPH. Norathyriol inhibited the iodonitrotetrazolium violet (INT) reduction in arachidonic acid (AA)-activated cell-free NADPH oxidase system at the same concentration range as those used in the suppression of PMA-activated particulate NADPH oxidase activity. Taken together, these results suggest that the scavenging ability of norathyriol contributes to the reduction of generated O2.-, however, the inhibition of O2.- generation from neutrophils by norathyriol is attributed to the blockade of PLC pathway, the attenuation of protein tyrosine phosphorylation, and to the suppression of NADPH oxidase through the interruption of electrons transport. PMID- 9358248 TI - Inhibitory effect of dipyridamole and its derivatives on lipid peroxidation in mitochondria. AB - Dipyridamole (DIP), 2,6-bis(diethanolamino)-4,8-dipiperidino-[5,4-d] pyrimidine, is a coronary vasodilator widely used in clinics. It has also been reported to have coactivator activity for a number of antitumour drugs and antioxidant activity in membrane systems. In recent years we have been studying the spectroscopic properties of this drug and several of its derivatives as well as their interaction with charged micelles and phospholipid monolayers. A strong interaction of DIP and DIP derivatives with these model membrane systems and a dependence of the strength of the interaction upon the chemical structure of the DIP derivative was observed. Here, the antioxidant effect of DIP and the derivatives, RA14, RA47, and RA25, was compared. We observed that although it strongly inhibits the iron-induced lipoperoxidation on mitochondria (IC50 = 1 microM), it shows no protection against an organic oxidant, cumene hydroperoxide. The order of hydrophobicity of the DIP derivatives, DIP > RA14 > RA47 > RA25, correlates very well with both the values of the association constants of these derivatives to micelles, their localization in the micelles, and phospholipid films and their antioxidant effect on mitochondria. So, a very good correlation of the structure of the drug in regarded to the nature of its substituents with the biological activity is observed. Essentially the same result was observed either measuring the lipid peroxidation or the membrane fluidity by ESR, suggesting that the effect of DIP and DIP derivatives is probably associated to their binding to the lipid bilayer and not to interaction with membrane proteins. PMID- 9358250 TI - Intermittent anoxia reduces oxygen free radicals formation during reoxygenation in rat hepatocytes. AB - The sensitivity of liver cells to anoxia is a major problem afflicting liver preservation and transplantation. Intermittent ischemia has been proposed to reduce reperfusion injury. The aim of the study was to assess oxygen free radical formation and cell injury during continuous or intermittent anoxia/reoxygenation in rat hepatocytes. Anion superoxide was measured by lucigenin-enhanced chemiluminescence and cell damage by LDH release and trypan blue uptake. During anoxia, superoxide generation dropped to background level in both groups; trypan blue uptake and LDH release, which increased progressively, were significantly greater in hepatocytes exposed to continuous compared to intermittent anoxia. During reoxygenation, a massive generation of superoxide anion formation, followed by a sharp increase in LDH release, was observed in both groups. However, both oxyradical generation and cell injury were significantly greater in cells exposed to continuous compared to intermittent anoxia. The data, showing that intermittent oxygen deprivation reduce liver cell injury and oxygen free radical formation determined by anoxia/reoxygenation, suggest a novel possible approach to the reduction of reperfusion injury. PMID- 9358249 TI - Decreased glutathione results in calcium-mediated cell death in PC12. AB - Neuronal damage in certain cellular populations in the brain has been linked to oxidative stress accompanied by an elevation in intracellular calcium. Many questions remain about how such oxidative stress occurs and how it affects calcium homeostasis. Glutathione (GSH) is a major regulator of cellular redox status in the brain, and lowered GSH levels have been associated with dopaminergic cell loss in Parkinson's disease (PD). We found that transfection of antisense oligomers directed against glutamylcysteine synthetase (GCS), the rate limiting enzyme in GSH synthesis, into PC12 cells resulted in decreased GSH and increased levels of ROS. Decreased GSH levels also correlated with an increase in intracellular calcium levels. Data from this study suggest that dopaminergic neurons are very sensitive to decreases in the internal oxidant buffering capacity of the cell caused by reductions in GSH levels, and that alterations in this parameter can result in disruption of calcium homeostasis and cell death. These results may be of particular significance for therapeutic treatment of PD, as those dopaminergic neurons that are spared in this disorder appear to contain the calcium binding protein, calbindin. PMID- 9358251 TI - Progressive effect of alpha-phenyl-N-tert-butyl nitrone (PBN) on rat embryo development in vitro. AB - In the present study we demonstrated the effects of the spin-trapping agent alpha phenyl-N-tert-butylnitrone (PBN) on the in vitro development of rat embryos at the early stage. In rat embryos, PBN increased the speed of the first cleavage and had no toxicity during pregnancy after embryo culture. These results showed that reactive oxygen species (ROIs) that were formed by activating molecular oxygens through redox reactions regulated the speed of development for early stage embryos. Thus, PBN caused a decrease in the level of ROIs and toxicity and an in increase in the level of the development of rat embryos. On the other hand, PBN could not decrease the 2-cell block in vitro nor increase the blastulation rate, in contrast to the fact that a scavenger of superoxide anions, SOD, is effective in doing so for mouse embryos. From these results it was concluded that free radicals play an important role in the in vitro development of rat embryos at the early stage, but play no role in the decrease of the 2-cell block or their blastulation rate. It should be noted that PBN had no toxicity for embryonic development at the 2-cell stage. PMID- 9358252 TI - Measurement of aldehydes in low density lipoprotein by high performance liquid chromatography. AB - A modified procedure is presented for the HPLC determination of nanomolar concentrations of n-alkanals, hydroxyalkenals, malondialdehyde and furfural in biological fluid. The modifications allow aldehyde profile analysis of small samples of fresh, human, low density lipoprotein (LDL), enabling more detailed studies of LDL fatty acid peroxidation. Aldehydes are reacted with 1,3 cyclohexanedione to produce fluorescent derivatives which are separated by gradient, reversed phase, high performance liquid chromatography (HPLC). Analysis time has been reduced by shortening the sample preparation. Sensitivity has been increased by miniaturization of the derivatisation procedure, reducing required sample size. Recoveries of added aldehydes have been improved. In addition, the method presented allows determination of three further aldehydes, not measured previously by CHD methods: malondialdehyde, formaldehyde and furfural. Recovery and variability data and concentrations of aldehydes found in human LDL are given. The capacity of the method for further development, to enable determination of other aldehydes such as the trans, 2-alkenals, is also demonstrated. PMID- 9358254 TI - Growing challenges to managed care. PMID- 9358253 TI - Absorbance changes of carotenoids in different solvents. AB - Carotenoids are typically measured in tissues with the high performance liquid chromatography (HPLC) and quantitation is usually done by calibrating with stock solutions in solvents. Four carotenoids including lutein, zeaxanthin, lycopene and beta-carotene were dissolved in hexane and methanol respectively, and their absorbance characteristics were compared. Lutein shows absorbance spectra that are almost independent of solvents at various concentrations. Spectra of zeaxanthin, lycopene and beta-carotene were found to be more solvent-dependent. The absorbance of zeaxanthin at lambda max is about approximately 2 times larger in methanol than in hexane at the higher concentrations, and increased non linearly with increasing concentration in hexane. The absorbance of lycopene at lambda max in hexane is approximately 4 fold larger than in methanol, but the absorbance of the methanol sample can be recovered by re-extracting this sample in hexane. The absorbance of beta-carotene in hexane is larger than in methanol, and increased linearly with increasing concentration. But beta-carotene showed a non-linear concentration effect in methanol. There are very small variations in lambda max for all four carotenoids between hexane and methanol, due to differences in molar extinction coefficients. The non-linear concentration effects for these carotenoids are probably due to differences in solubility leading to the formation of microcrystals. Thus, care should be taken with quantitation of tissue carotenoid values, when they depend on measurement of concentrations in stock solutions. PMID- 9358255 TI - Medical knowledge overload: a disturbing trend for physicians. PMID- 9358256 TI - Medical knowledge overload: a disturbing trend for physicians. PMID- 9358257 TI - Toward healthier hospitals. AB - This article builds around a framework of cure, care, control, and community, with collaboration at the center, to consider 12 issues common to many hospitals. These include, among others, the fragmentation of efforts, confusion in mission (and in mission statements), the problems of bundling research with clinical work, selectivity in informing board members, the dangers of professional management, and the difficulties of combining external advocacy with internal reconciliation in the senior manager's job. The article concludes that hospitals could better learn how to solve systemic problems systemically, and that to do so will require not the wish lists of strategic planning and structural reorganizing, but tangible changes in their collective behavior. PMID- 9358258 TI - Maintaining the new practice networks. AB - Unless acquired physicians are managed carefully, many will flee the acquiring networks in anger and frustration. Because older networks relied on a self selected population of physicians, the newer networks will have to develop alternative strategies to motivate those physicians who did not self-select for employment. This article makes recommendations about how to build a corporate practice culture under these new conditions. PMID- 9358259 TI - Physician practice management companies. AB - Physicians are increasingly courted by insurers and hospitals as partners for integrated delivery systems. A new integrative option has emerged for physicians- the physician practice management company (PPMC). PPMCs have formed in response to several supply and demand factors. They hold out great promise for physicians, but closer scrutiny suggests that this promise has not yet been realized. PPMCs warrant managerial and research attention due to their contracting potential as physician networks in dealing with employers and payers. PMID- 9358260 TI - Pure versus hybrid: performance implications of Porter's generic strategies. AB - This article identifies the strategic types in the hospital industry based on the hospital's use of Porter's generic strategies in their pure and hybrid forms. The article also examines differences in performance of hospitals across strategic types. Results indicate that hospitals that follow a focussed cost leadership strategy, in general, have superior performance on a variety of performance measures, while hospitals that use a combination of cost leadership and differentiation perform the poorest. Implications of findings for hospital administrators are also discussed. PMID- 9358261 TI - Skill-specific staffing intensity and the cost of hospital care. AB - Hospital managers have changed their staffing strategies to reduce the cost of care. Such hospitals may be both understaffed and underskilled in terms of some caregivers. Understaffing and underskilling may have indirect effects that offset the benefits of payroll cost reductions. This article indicates that in 1991 some California hospitals were understaffed in terms of specific bedside caregivers and had higher costs than did hospitals employing more of those caregivers. PMID- 9358263 TI - Hospital administrators' career paths: which way to the top? AB - This article examines career paths of aspirants to hospital administrator positions. It focuses on successful career objectives, barriers, and paths. The 1994 survey data from 162 hospital-employed executive track personnel in a western state facilitate comparisons with nonaspirants. Only 34 (21 percent) self reported a goal to become an administrator. Aspirants require institutional and mentor support, and need to follow more proactive and diverse career paths than they have done previously. PMID- 9358262 TI - Best practices for managing surgical services: the role of coordination. AB - Growing evidence exists that patient outcomes are related to how effectively health care organizations coordinate work responsibilities among their staffs. However, information is lacking on actual practices that can be used to achieve effective coordination. This article reports on a National Veterans Affairs Surgical Risk Study, in which the authors studied the coordination practices of 20 surgical services that, based on risk-adjusted mortality and morbidity rates, occupied different ends of the patient outcomes continuum. PMID- 9358264 TI - Trade-off between false positives and false negatives in the linkage analysis of complex traits. AB - This study examines the issue of false positives in genomic scans for detecting complex trait loci using subpair linkage methods and investigates the trade-off between the rate of false positives and the rate of false negatives. It highlights the tremendous cost in terms of power brought about by an excessive control of type I error and, at the same time, confirms that a larger number of false positives can occur otherwise in the course of a genomic scan. Finally, it compares the power and rate of false positives obtained in preplanned replicated studies conducted using a liberal significance level to those for single-step studies that use the same total sample size but stricter levels of significance. For the models considered here, replicate studies were found more attractive as long as one is willing to accept a trade-off, exchanging a much lower rate of false negatives for a slight increase in the rate of false positives. PMID- 9358265 TI - Self-contained subsets method for estimation of gene frequencies of truncated genetic data. AB - The self-contained subsets method subdivides a genetic data set into a number of subsets from which estimates are computed. The advantage of such a method is that when a subset is suspected of containing unreliable data then discarding that subset will not invalidate the remaining subsets for estimation. Thus, the complicated computation required to deal with truncated data can be avoided. In this paper, using estimation of gene frequencies as an example for one subset case, the marginal distribution of a subset total is derived and then, using this distribution, the variance of the frequency estimate of a gene frequency from the subdivision method is calculated. The subdivision method is also applied to impute the number of people of a truncated group. Finally, more complex cases where there are multiple subsets available for estimation are discussed. Results are compared to those of previous studies. PMID- 9358266 TI - Sample size required for predefined linkage decision quality. AB - A method for estimating the sample size required to attain a predefined linkage decision quality (type I and type II errors) is proposed using the linkage test power estimate developed by Ginsburg et al. [(1996) Genet Epidemiol 13:355-366]. The method is applicable for samples of arbitrarily structured pedigrees collected via proband. Comparison of different ascertainment schemes and pedigree structures by their consequent minimal sample size was performed. For recessive and dominant inheritance with complete penetrance, the relative ranks of the ascertainment schemes are invariant regardless of the true recombination fraction value and the trait and marker gene frequencies, which enables one to point out the better scheme. The feasibility of evaluating a sampling strategy by the cost of pedigree collection is also considered, and comparison between these two methods of sample planning is performed. PMID- 9358267 TI - Trends and patterns of mortality associated with birth defects and genetic diseases in the United States, 1979-1992: an analysis of multiple-cause mortality data. AB - Contemporary information on the trends and patterns of mortality associated with birth defects and genetic diseases is lacking in the United States. To study these trends and patterns, we used the Multiple-Cause Mortality Files of the National Center for Health Statistics. From 1979 through 1992, 320,208 deaths in the United States were associated with birth defects and genetic diseases. The age-adjusted mortality rates for people with birth defects declined from about 8.2/100,000 in 1979 to about 6.7/100,000 in 1992, and the mortality rates for people with genetic diseases increased from 2.2/100,000 in 1979 to 2.5/100,000 in 1992. The mortality rate was higher among men than among women and higher among blacks than among whites or other races for both birth defect- and genetic disease-associated deaths. The rate among infants with birth defects was more than 25 times higher than that among other age groups. About half of the children whose deaths were associated with birth defects had cardiovascular system defects, 15% had central nervous system defects, and 12% had chromosomal defects. For deaths associated with genetic diseases, hereditary neurologic or storage disorders were the most common genetic diseases (38%), followed by metabolic disorders (21%), sickle cell and thalassemia (12%). The decline in the rate of mortality from birth defects in the United States probably reflects improvements in medical and surgical care and other factors. Most of the mortality associated with birth defects remains in the pediatric age group (less than 15 years old). The upward trend we detected for the deaths with genetic diseases was most likely related to improved recognition and reporting of some genetic diseases rather than to the increased prevalence. PMID- 9358268 TI - Paternal age and sporadic neurofibromatosis 1: a case-control study and consideration of the methodologic issues. AB - Sporadic neurofibromatosis 1 (NF1) occurs in the absence of a family history of the disease and usually results from a new mutation in the germ cell of one of the parents, most commonly the father. Older paternal age may increase the risk for a new germinal NF1 mutation, but the results of studies to address this question conflict. We investigated paternal age in sporadic NF1 by using a case control study design. Patients who were seen at two specialty NF clinics in Houston, Texas, born between 1970 and 1992 and living in the Houston area and surrounding counties, were studied. Birth certificates with information on the father were found for 89 cases. For each case, two birth certificates were chosen at random from the same year and county of birth. In this way, the control group of 178 individuals was formed. Fathers of patients with NF1 were 1.5 years older than fathers of control subjects at the birth of the child, but the difference was only of borderline statistical significance (P = 0.07). This paternal age difference was not changed by adjustment for socioeconomic status or maternal age. These and previous data are consistent with either a small paternal age effect in sporadic NF1 or a bias such as that resulting from the selection of cases and/or controls. PMID- 9358269 TI - Therapeutic immunization against cancer antigens using genetically engineered cells. PMID- 9358271 TI - Displaying human interleukin-2 on the surface of bacteriophage. AB - Previous attempts to produce active human Interleukin-2 (hIL-2) in E. coli have failed, due to its aggregation in the form of cytoplasmic inclusion bodies, and the inability of the protein to enter the periplasmic export pathway, when fused to bacterial signal sequences. We have reasoned that these limitations could be overcome by introducing changes in the signal sequence and/or in some hIL-2 residues, not critical for its biological activity; and proceeded to test this hypothesis using a phagemid vector carrying the pelB secretion signal sequence, and the filamentous phage display system. Deletion of the Pro +2 in hIL-2 led to the export of a correct size (processed) molecule to the bacterial periplasm of Su- cells by the phagemid vector. However, this was achieved under growth conditions that would not favor phage assembly in Su+ strains. Changing the hydrophobic core of the leader peptide reversed this situation and allowed phage assembly and display of a pIII/hIL-2 hybrid protein in TG1 cells. The phage displayed hIL-2 is correctly folded, as judged by its ability to interact with a conformation-specific anti-hIL-2 monoclonal antibody, and maintains its biological activity when tested in a CTLL-2 cell proliferation assay. The changes introduced in hIL-2 and the signal sequence will make possible to use the powerful phage display technology for the selection of high-affinity variants from libraries of hIL-2 mutants. PMID- 9358270 TI - A TNF receptor antagonistic scFv, which is not secreted in mammalian cells, is expressed as a soluble mono- and bivalent scFv derivative in insect cells. AB - Single chain antibodies (scFv) are usually produced in E. coli, but generation of certain scFv derivatives, such as complex fusion proteins or glycosylated forms of scFv is restricted to eukaryotic expression systems. We investigated the production of soluble mono- and bivalent single chain antibodies (scFv) in eukaryotic cells and describe a cassette vector system for mammalian and baculovirus expression which is compatible with an established vector system for bacterial expression and phage display selection of scFvs. The applied model scFv was derived from a murine antibody (H398) against human tumor necrosis factor receptor 1 (TNFR60), known to be a potent antagonist of TNF action in its monomeric form and a potential therapeutic agent for treatment of TNF-mediated diseases. Surprisingly, the monomeric scFv form of H398 (scFv H398) is expressed but not secreted in different mammalian cells. In contrast, in insect cells using recombinant baculovirus, a monovalent scFv H398 and a bivalent scFv fusion protein with an human IgG1 Fc region were expressed and secreted with correctly processed signal sequence. Concerning the influence of valency of the model Ab and its derivatives on antigen binding affinity and neutralisation of TNF activity, we found that the mono- and bivalent form of scFv H398 possesses the same characteristics as proteolytically produced Fab H398 and original mAb H398, respectively. Furthermore, fusion of the Ig Fc protein to scFv H398 increase the in vitro half-life at 37 degrees C. We conclude that the described cassette vectors readily allow the eukaryotic expression of mono- and bivalent scFv derivatives to analyse the influence of valency of scFv molecules on antigen binding and biological activity. PMID- 9358272 TI - Evaluation of different lymphoid tissue sources for the construction of human immunoglobulin gene libraries. AB - BACKGROUND: Phage display technology allows the isolation of novel human monoclonal antibodies. The technology relies on the construction of a recombinant antibody library and its display on phage particles. The quality of an antibody library is affected by several factors including the size, diversity and source of immunoglobulin genes. OBJECTIVE: The aim of the project was to determine the best tissue source for the construction of antibody libraries. STUDY DESIGN: Three tissue sources were used in this study: peripheral blood mononuclear cells from a healthy donor, Epstein-Barr virus (EBV) transformed peripheral blood mononuclear cells and lymph node tissue from individuals with breast cancer. The quality of each tissue source was assessed using two criteria: (1) the number of mature and activated B cells in each source; (2) the amount of immunoglobulin heavy and light chain genes amplifiable by polymerase chain reaction (PCR). RESULTS: EBV-transformed peripheral blood mononuclear cells and lymph node tissue were shown to contain more B cells than peripheral blood mononuclear cells. A relatively larger amount of immunoglobulin gene products could be amplified from EBV-transformed peripheral blood mononuclear cells and the lymph node. However, immunoglobulin containing gamma 1 chains could not be amplified from EBV transformed mononuclear cells, and the resultant pattern of gene amplification suggests a possible selection bias. CONCLUSION: This study indicates that among the three tissue sources examined, lymph node tissue is the most suitable source for the construction of antibody libraries. PMID- 9358273 TI - Optimization of scFv antibody production in transgenic plants. AB - BACKGROUND: Plants offer various advantages for the production of pharmaceutical proteins over conventional production systems such as bacterial or mammalian cell culture. In order to explore transgenic plants for large-scale production and storage of recombinant antibodies we tried to optimize the accumulation and stability of functionally active single chain Fv (scFv) antibodies in transgenic tobacco plants. OBJECTIVES: Two different scFv antibodies which were expressed in different plant organs and plant cell compartments have been used for the study. Accumulation levels and antibody properties such as stability and antigen-binding activity were investigated. STUDY DESIGN: For ubiquitous expression in tobacco plants, transcription of the scFv genes was controlled by the strong cauliflower mosaic virus (CaMV) 35S promoter. We used seed specific legumin B4 (LeB4) and the unknown seed protein (USP) promoters from Vicia faba for storage organ specific expression. RESULTS: High accumulation of the two different scFv proteins in transgenic tobacco plants was only achieved by retention of the recombinant antibodies in the lumen of the endoplasmic reticulum (ER). Expression levels of scFv antibodies reached up to 4-6.8% of total soluble proteins (TSP) in leaves and up to 3-4% in ripe tobacco seeds. Transgenic tobacco seeds as well as tobacco leaves facilitated stable storage of ER-accumulated scFvs over an extended (seeds) or a short (leaves) period of time. Functionally active scFv proteins could be extracted after harvesting of the leaf material--drying and storage for 1 week at room temperature. Both the amount and the binding activity of the scFv proteins remained unchanged. CONCLUSION: A plant expression system where the scFv proteins are targeted in the ER provides not only the highest accumulation level of active single chain Fv antibodies ever reported but also a short- or long-term storage of the foreign protein in the harvested plant material. PMID- 9358274 TI - Regulation and expression of human Fabs under the control of the Escherichia coli arabinose promoter, PBAD. AB - BACKGROUND: The L-arabinose operon from E. coli contains an inducible promoter PBAD which has been extensively studied for the control of gene expression. PBAD has a number of potential advantages over Plac, and has been used successfully to promote high level expression of recombinant proteins. OBJECTIVES: The aim of this study was to investigate PBAD as an alternative system to Plac for the bacterial expression of recombinant Fabs. STUDY DESIGN: The promoter PBAD from the E. coli arabinose operon araBAD and the gene encoding the regulator of this promoter, were cloned into the phagemid expression vector MCO1. Expression of human recombinant tetanus toxoid (TT) and c-erbB2 Fabs under the control of PBAD was compared at two induction temperatures with the same Fabs produced under the control of Plac. RESULTS: Expression of TT and c-erbB2 Fabs under the control of PBAD was comparable to Fab expression from Plac. However, highly expressed TT Fab under the control of PBAD was localised to the soluble periplasmic fraction whereas under the control of Plac, there was greater leakage of Fab into the culture supernatant. In addition, Fab expression from PBAD could be more tightly repressed than from Plac. CONCLUSION: PBAD is a useful and cheaply inducible alternative to the more commonly used Plac for the rapid expression of soluble recombinant human antibody fragments. PMID- 9358275 TI - Effects of antisense hsp27 gene expression in osteosarcoma cells. PMID- 9358276 TI - A simplified method for tissue engineering skeletal muscle organoids in vitro. PMID- 9358277 TI - Chromosomal analysis of two established salmonid cell lines: CHSE-214 (Oncorhynchus tschawytscha) and RTG-2 (Oncorhynchus mykiss) PMID- 9358278 TI - Efficient introduction of genes into human ovarian surface epithelium. PMID- 9358279 TI - Establishment of uterine cell lines from p53-deficient mice. PMID- 9358281 TI - Irradiation of murine donor spleen cells with ultraviolet "B" light eliminates graft-versus-host and host-versus-graft response in allogeneic recipient mice. PMID- 9358280 TI - Cytotoxicity of beta-amyloid peptide 25-35 on vascular smooth muscle cells and attenuation by vitamin E. PMID- 9358283 TI - Sarcomatous and adenocarcinoma cell lines from the same nodule of cholangiocarcinoma. PMID- 9358282 TI - A primary cell culture model for defective cardiac myofibrillogenesis in Mexican axolotl embryos. PMID- 9358284 TI - An in vitro three-dimensional coculture model of cerebral microvascular angiogenesis and differentiation. AB - The microvasculature of the developing brain is plastic and responds differently to the many insults associated with preterm birth. We developed three-dimensional in vitro culture models for the study of the responses of the developing cerebral microvasculature. Beagle brain microvascular endothelial cells (BBMEC) were isolated by differential centrifugation from newborn beagle pups on postnatal Day 1 and placed in three-dimensional culture dispersed in a collagen gel. Alternatively, BBMEC were placed in a three-dimensional coculture with neonatal rat forebrain astrocytes. Cultures were analyzed for extracellular matrix components at 1 and 6 d, and total RNA was extracted for Northern analyses. Urokinase plasminogen activator activity was assayed in both mono- and cocultures of the two cell types. Studies of three-dimensional BBMEC/astrocyte cocultures demonstrated progressive tube formation with only low levels of endothelial proliferation. By 6 d in three-dimensional coculture, the BBMEC formed capillarylike tubes with a wrapping of glial processes, and basement membrane protein synthesis was noted. Urokinase plasminogen zymography suggested intercellular signaling by the two cell types. These data suggest that the three dimensional beagle brain germinal matrix microvascular endothelial cell/neonatal rat astrocyte coculture provides a good model for the investigation of microvascular responses in the developing brain. PMID- 9358285 TI - Morphological and biochemical characterization of primary culture of rabbit proximal kidney tubule cells grown on collagen-IV coated Millicell-CM. AB - The aim of this study was to better characterize rabbit proximal kidney tubule cells cultured on collagen IV-coated porous inserts, as compared to the same cells seeded in standard plastic wells. Total protein contents in confluent monolayers on permeable membranes were about twofold higher than those measured in confluent cultures in plastic wells. Microscopy examinations suggested that such a difference was probably due to a higher cell density and to an impressive development of the apical brush-border membrane. Moreover, measurement of unidirectional transport of p-aminohippuric acid and tetraethylammonium bromide confirmed the high polarization level of cultures on porous inserts. Results of methyl(alpha-D-[U-14C]glyco)pyranoside uptake suggested that cell phenotype was probably influenced by culture conditions. Analysis of different markers as a function of time in culture showed decreases of alkaline phosphatase (AP), gamma glutamyltranspeptidase (GGT), and Na(+)-K(+)-ATPase activities as well as increases in LDH, ATP, and glutathione levels, similar to those formerly reported for cells cultured in standard plastic plates. However, comparative data from 6-d old monolayers have shown that AP, GGT, Na(+)-K(+)-ATPase, glutathione reductase (GRED), and selenium-dependent glutathione peroxidase (Se-GPX) activities were 2.8-, 2.6-, 1.6-, 1.2-, and 2.1-fold, respectively, better preserved on precoated permeable membranes. On the other hand, this paper reports for the first time in the literature that GRED and SE-GPX, two phase II detoxification enzymes, were well maintained in cultures of rabbit proximal kidney tubule cells. Our results show that culturing rabbit proximal kidney tubule cells on collagen IV-coated porous membranes was accompanied by an improvement of both morphological and biochemical properties of the cells. PMID- 9358286 TI - Restoration of the differentiated functions of serially passaged chondrocytes using staurosporine. AB - Among the various directions explored in order to have a large number of differentiated articular chondrocytes easily available, the restoration of the differentiated properties after cell multiplication in monolayer has been proposed. It has been clearly shown that the synthesis of cartilage proteoglycans and type II collagen synthesis is coincident with the presence of a faint microfibrillar architecture but is absent in chondrocytes showing well-defined actin cables. Staurosporin, mainly described as a protein kinase C inhibitor, has also been shown to rapidly induce the disruption of the actin microfilaments. The purpose of this paper was to investigate whether properties of differentiated chondrocytes were reinitiated upon staurosporin treatment of serially passaged chondrocytes. Results showed, after staurosporine treatment of cells at Passage two for 5 d, complete suppression of type I and type III collagen synthesis and induction of type II collagen synthesis and of Alcian blue stainable matrix. Additionally, we showed that staurosporin restored metabolic responses that chondrocytes in primary culture exhibit upon interleukin-1 beta treatment (decrease of Alcian blue- positive cells, induction of expression of the 92 kDa gelatinase, nitric oxide production). We conclude that staurosporin is a potent redifferentiating agent of articular chondrocytes that have been subcultured up to Passage two for multiplication. Taking into account that the cellularity of cartilage is very low, staurosporine-treated chondrocytes could be useful as an alternative cellular model to evaluate pharmacotoxicological effects of drugs. PMID- 9358287 TI - Effect of a mistletoe extract (Iscador QuFrF) on viability and migratory behavior of human peripheral CD4+ and CD8+ T lymphocytes in three-dimensional collagen lattices. AB - Migration and tissue distribution of immunocompetent cells may be critical prerequisites for efficient immune surveillance. The effect of various concentrations of the mistletoe extract Iscador QuFrF on the locomotory behavior and viability of immunomagnetically isolated human CD4+ and CD8+ T lymphocytes within three-dimensional collagen gels was investigated. Although variation in baseline activities of spontaneously migrating T cells was donor-dependent, a dose-dependent stimulation of the locomotory activity in both CD4+ and CD8+ T cells for noncytotoxic concentrations of Iscador QuFrF (0.25-1.25 micrograms/ml) was detected. The optimal concentration of mistletoe extract and time of maximal response were specific for each donor. As shown by cell tracking and subsequent data analysis, CD4+ T cells exposed to the mistletoe extract displayed a significant increase in mean velocity and time locomoting; total distance migrated was nearly doubled. In contrast, CD8+ T cells showed less pronounced changes in these critical parameters. Cytotoxic effects of the mistletoe preparation on T lymphocytes, which could at least partially be attributed to the induction of apoptosis, were drastically reduced in the presence of fetal calf serum in the culture system. Our data suggest that the direct stimulation of T cell migration in the presence of mistletoe components may modulate in a dose dependent manner the system of immune surveillance and recognition in patients under mistletoe therapy. PMID- 9358289 TI - To sleep, perchance to dream. PMID- 9358290 TI - The state of adolescent health: looking back and planning ahead. AB - PURPOSE: A decade has passed since the 1986 Health Futures of Youth policy-making conference was held. The present study aimed to examine perceived changes in the field of adolescent health since this conference, as a basis for further planning in adolescent health and consideration of a possible follow-up conference. METHODS: The study included two parts: (a) a mailed survey which was completed by 68 of 90 conference participants; and (b) telephone interviews with a sample of leaders from federal agencies and major foundations and one professional organization serving youth in the United States. RESULTS: Respondents perceived that small to moderate improvements have occurred for 10 key recommendations made at the 1986 conference. There appears to be increased recognition of the special needs of youth and a greater understanding of the period of adolescence. Improvements in health-related activities and in research of health behaviors have occurred. However, perceived setbacks have occurred at the intersection of health and social welfare. The conference was viewed by respondents as having had a moderate influence on the field of adolescent health and on the participants' own work and working relationships. In addition, policy-making conferences were listed as the strongest source of influence on the agenda of national agencies and foundations serving youth. CONCLUSION: A follow-up conference is recommended and should focus on both health-related topics and issues of social welfare. PMID- 9358288 TI - Establishment and immunocharacterization of an immortalized pancreatic cell line derived from the H-2Kb-tsA58 transgenic mouse. AB - This study describes the establishment and characterization of an immortalized cell line derived from the pancreas of an adult H-2Kb-tsA58 transgenic mouse. These cells, designated IMPAN for IMmortalized PANcreatic cells, displayed a cobblestone appearance typical of confluent epithelial cells and a distinct polarity in the organization of their cytoplasmic organelles. Immunocytochemical studies revealed that all IMPAN cells stained positively for a wide range of markers characteristic of pancreatic acinar cells, namely the secretory products alpha-amylase, chymotrypsinogen, DNAse, the lectinlike secretory protein PAP (pancreatitis associated protein), and the zymogen granule membrane proteins GP-2 and gp300. They also stained positively for carbonic anhydrase II and cytokeratin 19, two proteins characteristic of pancreatic duct cells, as well as for rab3A, a small GTP-binding protein specifically localized in pancreatic islet cells. No reactivity was ever obtained with insulin antibodies. Taken together, these results show that the IMPAN cells exhibit a phenotype comparable to exocrine pancreatic acinar cells. However the expression of some proteins more specific to duct and islet cells make them similar to in vivo or in vitro growing acinar cells. The cell line should be a valuable model to study the mechanisms of growth, differentiation, and transformation of the exocrine pancreatic acinar cell. PMID- 9358291 TI - Barriers to health care for street youth. AB - This study investigates the barriers to health care faced by runaway adolescents. A convenience sample of 89 street youth located through community agencies was surveyed to elicit their perceptions of barriers to care. Results indicated that these youth experience a wide range of barriers to health care, both objective and subjective. They also experience fears with regard to receiving health care, many of which seem developmental in nature. The relative isolation of these youth compounds the objective barriers they face, yet many overcame these barriers and received needed care. PMID- 9358292 TI - Community and dating violence among adolescents: perpetration and victimization. AB - PURPOSE: Adolescents are both the perpetrators and victims of violence in the United States. To reduce violence, it is important to identify those most at risk within particular contexts. METHODS: A social learning framework was used to investigate involvement in violence in a survey of 719 high school students. Four outcomes (community violence perpetration, community violence victimization, dating violence perpetration, and dating violence victimization) were examined as a function of demographic characteristics, exposure to violence, and several potential mediating variables. RESULTS: Exposure to weapons and violent injury in the community was the sole consistent predictor across the four outcomes. Gender generally was an important correlate of violence; there were substantial gender differences in the correlates of dating violence perpetration and victimization, but relatively few gender differences in the correlates of community violence involvement. Other demographic characteristics typically were of limited importance, and were largely accounted for by exposure to violence or other mediators. Personal norms about the circumstances under which the use of violence is perceived as justified were important for three of the four outcome: community violence perpetration, and dating violence perpetration and victimization. CONCLUSIONS: Being exposed to violence in one context appears to have crossover effects to victimization and perpetration in another context. Furthermore, victimization and perpetration often co-occur. PMID- 9358293 TI - Factors associated with aggressive and delinquent behaviors among patients attending an adolescent medicine clinic. AB - PURPOSE: The purpose of this study was to examine the associations among delinquency/aggressiveness and alcohol or drug use, sexual risk behaviors, and other scales from the Problem-Oriented Screening Instrument for Teenagers (POSIT). METHOD: A total of 173 15- to 18-year-olds attending adolescent medicine clinic completed a 183-item questionnaire. The questionnaire contained the POSIT, the CAGE questions, a 22-item alcohol and drug screen (AOD), and a human immunodeficiency virus and sexual risk behavior screen. The dependent variable was the Aggressive Behavior/Delinquency (ABD) scale from the POSIT. RESULTS: There were no gender or racial differences in the ABD scale (p > 0.05). The ABD scale was significantly (p < or = 0.001) associated with the POSIT AOD scale (r = 0.46), the 22-item AOD drug screen (r = 0.57), the CAGE questions (r = 0.39), the sexual risk behavior scale (r = 0.39), and the POSIT physical health (r = 0.34), mental health (r = 0.51), family relations status (r = 0.37), peer relations (r = 0.57), educational status (r = 0.44), social skills (r = 0.31), and leisure/recreation (r = 0.16) scales. Multiple regression analysis indicated that the AOD screen and problems in the area of peer relations, family relations, and mental health accounted for 47.7% of the variation in the ABD scale (p < or = 0.00001). CONCLUSIONS: These findings suggest that adolescents who engage in more aggressive and delinquent behaviors are more likely to use alcohol and other drugs, engage in sexual risk behaviors, report problems with peer and family relationships, and report more mental health symptoms. PMID- 9358294 TI - Preventing the spread of AIDS in youth: principles of practice from 11 diverse projects. AB - PURPOSE: The purpose of this study was to identify the factors that contributed to intervention effectiveness in acquired immunodeficiency syndrome (AIDS) prevention projects targeting youth. METHODS: Eleven AIDS prevention projects funded by the Robert Wood Johnson Foundation whose target populations consisted of at least 60% youth were studied. A blended methodology resulted in quantitative data (i.e., survey responses) from all 11 projects supplemented with qualitative data (i.e., open-ended interviews) drawn from in-depth site visits to six projects. RESULTS: Projects reported using a mean of 16.6 intervention activities (selected from a list of 30). Six activities were used by all 11 projects. Small group discussions were rated as one of three most effective activities by 72.7% of the projects that used them. Project staff identified three elements of effective interventions: involvement of peer educators, recognition of the role of adults (e.g., parents, teachers), and use of structured settings to gain access to the target population (e.g., schools, clubs). CONCLUSIONS: The most powerful strategies described by project staff for reaching adolescents at risk for human immunodeficiency virus (HIV) transmission also bring considerable challenges. Opportunity costs associated with using peer educators, gatekeeper support, and structured settings may include limited control of the message, impaired credibility, and failure to reach those youth at greatest risk of HIV infection, respectively. Health educators will do well to consider the advantages and disadvantages of these factors when developing, implementing, and evaluating AIDS prevention programs for youth. PMID- 9358295 TI - Predictors of reported condom use in central Harlem youth as conceptualized by the health belief model. AB - PURPOSE: To examine the relationship of reported condom use to specific sociodemographics, psychosocial variables, and perceptions of and motivations for condom use as conceptualized by the Health Belief Model. METHODS: This study performed a cross-sectional survey of 557 adolescents enrolled in a hospital based pregnancy prevention program in an urban community hospital (Harlem Hospital). Multiple logistic regression analysis examined the combined relationship of the significant psychosocial variables to consistent condom use. RESULTS: Males were less likely than females to report teen-parent conflict and depression and more likely to report support for birth control, participation in community activities, and favorable attitudes toward delaying parenthood. Consistent with the Health Belief Model adjusting for age, the strongest predictors of consistent condom use were partner preference for condoms, perceived benefit of avoidance of pregnancy, male gender, and support for birth control (usually by a parent). CONCLUSIONS: The data on this urban, predominantly African-American sample of adolescents suggest the importance of the influences on specific motivations to use protection--that is, the wish to avoid pregnancy, human immunodeficiency virus/acquired immunodeficiency syndrome, and sexually transmitted diseases, although the mechanisms are still unclear. In addition, gender and the modifying effects of parental and partner support of the use of protection strongly influence the reported use of condoms by adolescents. These factors (in addition to psychosocial factors such as depression) may be important in planning interventions to increase condom use by sexually active teens. PMID- 9358296 TI - Role of school-based health centers in referral completion. AB - PURPOSE: This study examined referrals from School-based health centers (SBHCs) to a sponsoring hospital to determine factors influencing successful referral completion and to assess SBHCs' ability to coordinate care. METHODS: A total of 138 referrals from eight SBHCs to Boston City Hospital between September 1993 and October 1994 were reviewed via medical records, clinic logs, and hospital registration system. A data extraction tool was used to collect information. Statistical analyses were performed to identify associations between referral completion and study variables. RESULTS: Seventy-five percent of all referrals were completed: 55.4% on the first attempt. Forty-six percent of those referred a second time completed their referrals. Statistically significant associations between referral completion and reason for referral (p = 0.01), visit diagnosis (p = 0.005), and usual source of health care (p = 0.009) were found. Provider documentation, including referral log and patient chart, was also associated with referral completion. Neither gender, race, nor health insurance had any significant association with successful referrals. CONCLUSIONS: This study suggests that SBHCs can significantly contribute to coordinated care for adolescents, especially for the uninsured and those without a source of health care. SBHCs were particularly effective at facilitating referral to specific services including: family planning, tuberculosis prophylaxis, and subspecialty care. Provider action, such as making appointments and documentation, was also an important factor. PMID- 9358297 TI - Adolescents' and providers' perspectives on the need for and use of mental health services. AB - PURPOSE: This study examines need for and use of services from both the adolescent's and the service provider's viewpoints. METHODS: The Youth Services Project interviewed 792 youths from the juvenile justice, education, primary health care, and child welfare sectors (200 each); gathered anonymous tallies of the mental health of youthful clients at each sector; and conducted focus groups with providers. RESULTS: A high percentage of youths (12-15%) met DSM-IV criteria for a mental health disorder, yet the sector clients were not identified as having mental health problems. Juvenile justice and child welfare sectors identified the highest percentage of adolescent clients as having mental health problems, and provided the most services (50-80%). The primary health care sector recorded no mental health disorders among the tallied clients, and provided the fewest mental health services (< 20%). Providers' complaints that they lacked knowledge concerning mental health assessment and lacked referral or treatment resources closely paralleled the degree to which their sector underserviced youths. CONCLUSION: Lack of knowledge about the extent of need in adolescents, methods for assessing or treating, and referral resources handicap service providers and explain the gap between need and service. PMID- 9358299 TI - Overview of ethical issues perceived by allied health professionals in the workplace. AB - Allied health professionals comprise a major segment of the total health care workforce. Recent advances in technology combined with increasingly complex patient-related issues require health care practitioners to be aware of and sensitive to bioethical dilemmas. This research assesses how allied health personnel perceive ethical issues in health care, and depicts their involvement in ethical situations. A combined qualitative and quantitative methodology was employed with thirty-six case study subjects at a large mid-west urban medical center. Two-thirds of the population were female and more than half of the sample had formal training in bioethics. Several ethical themes emerged in the workplace with allied health practitioners including issues of team-work, confidentiality, assessment, and documentation. Professional judgment was expressed as essential by all of the subjects. The allied health professionals perceived the need for structured coursework as a link between formal training and the health care setting to maintain the dynamics of the provider-patient relationship. PMID- 9358298 TI - Epidemiologic study of sleep quality and troubles in French secondary school adolescents. AB - BACKGROUND: The purpose of the study was to assess the prevalence and correlates of sleep problems in adolescents. METHODS: A total of 763 students chosen at random among the 15 secondary schools of a French departement were given a self report questionnaire. RESULTS: As much as 40.8% reported at least one of the five sleep disturbances we studied including difficulties falling asleep and staying asleep, a need for more sleep, early awakenings, and chronic sleeping pill intake. These sleep problems were highly related to various personal and family disorders. Discriminant analysis for categorical data pointed to a consistent but not significant descriptive profile accounting for sleep problems in these students. Suicide, weight concerns, and stimulant abuse were the most informative personal correlates as parts of this profile. CONCLUSION: These findings suggest that the complaint of poor sleep should be regarded with special care in adolescents as a possibly meaningful and sensitive sign of severe family or personal disruption. PMID- 9358300 TI - Practice environment and the employment of nurse practitioners, physician assistants, and certified nurse midwives by community health centers. AB - This report examines the relation between state variations in the regulation of nurse practitioners (NPs), physician assistants (PAs), and certified nurse midwives (CNMs), and the employment of these nonphysician providers (NPPs) by community health centers (CHCs). Data for this report came from a 1991-92 survey of CHCs assessing the employment of NPPs, and secondary available data. The dependent variables examined were the numbers of NPPs currently employed by CHCs. Independent variables included 1992 practice environment scores, CHC location, number of CHC physicians, and NPP-to-population ratios. The number of NPs and PAs employed by CHCs was significantly associated with practice environment for these practitioners. NPP-to-population ratios and the number of CHC physicians are also significantly associated with NPP employment by CHCs. State decision makers may reduce legislative and regulatory barriers to practice as a way to improve the practice environment for nonphysician primary care providers, particularly NPs and PAs. Thus, community health centers can employ adequate number of NPPs to fulfill their mission of serving the poor and underserved population. PMID- 9358301 TI - A process of developing terminal competencies for an interdisciplinary training program. AB - This article describes the process used in developing terminal trainee competencies for an interdisciplinary training program for professional students representing 13 disciplines. The major steps of the process are delineated and described. Although the process is explained in terms of its use in this specialized training program, it can be applied to other training programs that are interdisciplinary in their focus. PMID- 9358302 TI - Accreditation, core curriculum and allied health education: barriers and opportunities. AB - There is considerable discussion today about the future directions of accreditation of allied health education. A number of discussions and public debates have taken place in recent years which consider and speculate on appropriate directions, how these might be achieved, and what the ramifications will be for allied health education. Current attention to core curriculum in allied health has implications for accreditation of such curricula. This paper reviews the definition and purpose of accreditation, addresses the current accreditation system in allied health, summarizes multiple perspectives in the current debate over the future of allied health accreditation, and examines how accreditation might help or hinder development of core curricula. A set of accreditation standards for evaluating a core curriculum in allied health is proposed. PMID- 9358303 TI - Educating rural allied health professionals: an interdisciplinary effort. AB - This article describes the process and outcome of an innovative, interdisciplinary educational effort which was initiated in rural Maine. Within the context of a comprehensive, statewide interdisciplinary rural health care training project that was federally funded through the Interdisciplinary Training for Health Care for Rural Areas (ITHCRA) grant program, a primary strategy for recruiting and training rural allied health practitioners was the development and implementation of a year long interdisciplinary graduate course sequence. The results of the evaluation suggested that interdisciplinary education is not only useful in learning the roles and functions of allied health professions, but is valuable tool in promoting the development of skills necessary for competent and relevant rural allied health practice. PMID- 9358304 TI - Allied health and physician assistants: a progressive partnership. AB - The recently published report of the National Commission on Allied Health stressed the perceived need for the development of more multi-skilled, generalist Allied Health professionals as a means of responding to the primary care focus of health care in the United States. There are many significant barriers to the development of such a professional in terms of educational requirements and licensing among other issues. Physician assistants have fulfilled this role in health care for nearly 30 years. The relationship between Allied Health and physician assistant education is not always clear although many Allied Health professionals pursue further education as physician assistants. Further development and clarification of this relationship presents an opportunity for Allied Health to participate in the training of generalist practitioners without complicating the educational requirements of other Allied Health professional programs. PMID- 9358305 TI - Influence of delayed injection time on the creep behavior of acrylic bone cement. AB - It has been proposed that creep of acrylic bone cement may contribute to loosening of cemented total joint replacements. If true, it is important that factors affecting creep of bone cement are identified. The objective of this study was to evaluate the effect of an operator-controlled variable, injection time, on the creep behavior of acrylic bone cement. Results from this investigation showed that injection time significantly (p < 0.0001) influenced creep behavior of bone cement. "Delayed" injection time of acrylic bone cement increased creep by approximately 5 times in 24 h compared to specimens prepared according to standard injection procedures. PMID- 9358307 TI - Fracture and material degradation properties of cortical bone under accelerated stress. AB - The fracture stress and material property degradation of bovine cortical bone specimens were investigated experimentally under accelerated cyclic tensile stress testing. The fracture stress of a typical specimen was found from a static tensile test, and the cyclic loading/unloading was calculated as a percentage of this fracture stress. The results of accelerated cyclic stress tests were compared to monotonically increased static tests to determine if loading/unloading has an effect on the damage mechanism in bone. It was found that fracture stress of the bone increases due to accelerated stress cycling whereas the modulus decreases in a logarithmic fashion with increasing cyclic stress. PMID- 9358306 TI - Selective in vitro removal of anti-A antibodies by adsorption on encapsulated erythrocyte-ghosts. AB - Large volume plasma exchanges are used for the removal of anti-A or anti-B antibodies from the plasma of patients undergoing transplantation from donors with major ABO incompatibility. Previous works suggest that solid-phase immunoadsorption can be substituted for plasma exchange in situations where antigens can be purified and immobilized on columns through which plasma is percolated. However, the preparation of purified antigens of the ABO system is large quantities is laborious and requires the use of considerable blood volumes. Studies were therefore undertaken to determine the feasibility of an original immunoadsorbent based on porous microparticles prepared by a water/oil/water emulsification-solvent evaporation method, within which erythrocytes-ghosts carrying blood group antigens were entrapped. The decrease of the antibody hemagglutinating titre after adsorption onto encapsulated ghosts suggests that antibodies can cross the polymeric membrane and bind to the antigens. This original approach of using encapsulated antigens for the batchwise removal of antibodies could be extended to affinity chromatography, and immunoadsorption therapy with a chromatographic column linked to an extracorporeal circulation could be considered. PMID- 9358308 TI - The expression of integrin subunits alpha 6 and beta 4 by corneal epithelial cells on modified hydrogel surfaces. AB - Our goal was to quantitate the expression and localization of integrin subunits alpha 6 and beta 4 by corneal epithelial cells on defined synthetic substrates. Previously we demonstrated that the cytoplasmic pH and translocation of the alpha 6 integrin subunit to the cell membrane was modified by ionic interactions. These results suggest that changes in the ionic interactions at the cell-substrate interface not only alter the intracellular milieu but ultimately affect the expression of adhesion proteins. To test this hypothesis, hydroxyethylmethacrylate (hema) hydrogels were modified by the addition of amines (N,N-dimethylaminoethylmethacrylate) or carboxyl moieties (methacrylic acid). Changes in the distribution of mRNA and protein were monitored using confocal laser scanning microscopy. The steady state level of integrin mRNA was evaluated, and the results indicate that while the plating efficiency was identical on all surfaces, the expression and localization of integrin subunits was surface dependent. Alpha 6 and beta 4 proteins were localized along the basal surface of nonpermeabilized cells cultured on laminin, on surfaces with amine moieties, and on those with amine and carboxyl moieties. The level of diffuse cytoplasmic staining increased with the presence of carboxyl moieties. Alpha 6 and beta 4 integrin subunits were negligible when the surfaces contained carboxyl moieties alone. The expression of alpha 6 and beta 4 mRNA was higher on surfaces containing amine moieties than on surfaces containing only carboxyl moieties. These results indicate that the characteristics of the substrate and the resulting cell-matrix interaction alter protein and mRNA expression of integrin subunits. PMID- 9358309 TI - Flow cytometric analysis of material-induced platelet activation in a canine model: elevated microparticle levels and reduced platelet life span. AB - Assessment of material-induced platelet activation is important given that it is thought to be a major mechanism of biomaterials thrombogenicity. We monitored, by flow cytometry, platelet microparticle (MP) levels in the circulation during the connection of polyvinyl alcohol (PVA) hydrogel and polyethylene (PE) test segments (3.18 mm ID, 20 and 50 cm L) to our chronically shunted beagle dogs. We report that circulating microparticle levels were dependent on test segment material, length, and time. The connection of 50-cm lengths of PVA hydrogel test segments led to MP levels two to three times greater than background at 48 h, while the connection of polyethylene test segments did not lead to elevated microparticle levels. MP levels were near background 24 h after removal of the PVA test segment. To determine platelet life span during the connection of test segments, platelets were labeled in vivo with biotin and their disappearance monitored flow cytometrically. While platelet life span for shunted dogs (no test segment) was 4.7 +/- 0.2 days, the connection of PVA hydrogel test segments led to a platelet life span of < 2 days. PMID- 9358310 TI - Incorporation of nitric oxide-releasing crosslinked polyethyleneimine microspheres into vascular grafts. AB - Over the years, many attempts have been made to increase the patency of small- to medium-sized prosthetic vascular grafts. However, none of them has greatly affected long-term rates. Recently, nitric oxide (NO) has been shown to inhibit thrombus formation in such grafts, suggesting that local delivery of NO may help to increase graft patency. This study describes the site-specific delivery of NO by entrapping NO-releasing microspheres in the pores of a vascular graft. NO releasing polyethyleneimine microspheres (PEIX) were developed using a novel water-in-oil emulsion technique involving chemical crosslinking with a bis epoxide. The PEIX microspheres were then derivatized with NO forming the [N(O)NO] moiety of the diazeniumdiolates formerly known as NONOates. These polymeric NO releasing particles were found to spontaneously release 194 nmol NO/mg with a half-life of over 66 h under physiologic conditions. Fluorescein isothiocyanate labeled microspheres were then embedded into the pores of a 60-micron nonreinforced Gore-tex vascular graft using a simple evacuation technique and evaluated for microsphere placement and NO release. Scanning electron microscopic analysis showed the microspheres entrapped in the pores of the vascular graft releasing 10 nmol NO/mg with a half-life of 51 h. The microspheres remained entrapped in the graft even after immersion and NO release, as confirmed by fluorescence of the medium. These results suggest that NO-releasing particles can be incorporated into the pores of a vascular graft to deliver therapeutic amounts of NO for the prevention of thrombosis in small-diameter prosthetic grafts. PMID- 9358311 TI - Enhancement of artificial juxtacrine stimulation of insulin by co-immobilization with adhesion factors. AB - Insulin was co-immobilized with a cell adhesion factor--fibronectin or polyallylamine--on a surface-hydrolyzed poly(methyl methacrylate) film. Chinese hamster ovary cells overexpressing human insulin receptors were cultured on the film in the absence of serum or soluble proteins. While insulin immobilization did not affect cell adhesion, insulin immobilized on fibronectin-immobilized film reduced the adhesion. Addition of the tetrapeptide Arg-Gly-Asp-Ser (RGDS) inhibited cell adhesion onto fibronectin-immobilized films while cell adhesion onto polyallylamine-immobilized films was not inhibited by RGDS. Small amounts of immobilized insulin (1 to 10% of the amount of free insulin required to achieve cell growth acceleration) were sufficient to stimulate cell proliferation. The maximal mitogenic effect of immobilized insulin was greater than that of free insulin. In addition, co-immobilization with the adhesion factor remarkably enhanced the mitogenic effect. The phosphorylation of the receptor with free insulin attained the maximum degree very rapidly but ceased quickly. On the other hand, the receptor phosphorylation with immobilized insulin was accompanied by a longer induction period and lasted a longer period of time than that with free insulin. Insulin co-immobilization on fibronectin or polyallylamineimmobilized films reduced the induction period by enhancement of cell adhesion. The early and long-lasting receptor activation might have been caused by the greater mitogenic effect of co-immobilized biosignaling polypeptides. PMID- 9358312 TI - Organic surface chemistry on titanium surfaces via thin film deposition. AB - In order to develop a synthetic strategy for the fine tuning of the interfacial properties of titanium-based implants and implant parts, a thin polymeric film was deposited from ethylene plasma on the surfaces of Ti foils. The intended aim was to further modify the adherent, delamination-resistant organic coating using the techniques of surface modification of polymers to direct interfacial interactions at the metal foil-biological phase interface. In particular, air plasma treatment and Ce(IV)-induced hydroxyethylmethacrylate grafting, two typical reactions of biomedical polymers surface chemistry, were used to improve cell adhesion or to impart cell resistance to the plasma-coated Ti. Results indicate that a plasma-deposited thin polymeric film effectively can act as a viable substrate for further surface chemical modifications and allow the application of a huge background of surface-modification polymers to metallic devices. PMID- 9358313 TI - Radiochemical studies on contact lens soilation. II. Lens uptake of cholesteryl oleate and dioleoyl phosphatidylcholine. AB - Employing an artificial tear preparation composed of six proteins and six lipids as a deposit model, uptake of the lipids 3H-cholesteryl oleate and 14C-dioleoyl phosphatidylcholine was measured on contact lenses representative of the four FDA hydrogel groups and on select RGP lenses. Cholesteryl oleate uptake after 24 h at 37 degrees C generally was less than 1 microgram/lens although occasionally reaching 1-2 micrograms. DuraSoft 3 lenses (Group IV) accumulated the deposits in greater amounts (p = 0.04) with other lens groups not differing significantly from each other. Ionic DuraSoft 2 and 3 lenses bound more phosphatidylcholine (also < 1 microgram) than other lens groups, possibly reflecting an interaction between the positively charged choline residue and the negative surface of the lens. Lysozyme deposition, measured simultaneously with cholesteryl oleate, bound to a far greater extent to Group IV lenses (e.g., DuraSoft 3, mean surface deposit 279 micrograms) than to other lens types (p < 0.01). Multiple application of the artificial tear solution did not produce a statistically significant increase in cholesteryl oleate accumulation. PMID- 9358314 TI - Chemical inactivators as sterilization agents for bovine collagen materials. AB - The use of collagen as a biomedical implant raises safety issues with regard to viruses and prions. Specific chemical agents that inactivate prion infectivity could be applied to collagen implants. The physicochemical changes and the in vitro and in vivo biocompatibility of collagen treated by formic acid (FA), trifluoroacetic acid (TFA), tetrafluorethanol (TFE), and hexafluoroisopropanol (HFIP) were investigated. In addition, the effects of these treatments on nucleic acids incorporated in collagen were analyzed. The molecules of FA and, more important, of TFA remained within collagen. FA, TFA, and HFIP treatments modify the secondary structure of collagen, as shown by Fourier transform infrared spectroscopy, while TFE does not. Differential scanning calorimetry measurements showed a decrease in the denaturation temperature compared to untreated collagen. However, resistance to collagenase was modified only after HFIP treatment. In vitro, cell growth was not impaired; in vivo, implants induced a temporary inflammatory reaction that was prolonged with TFA and HFIP treatments. TFE and FA treated collagen were thoroughly infiltrated by fibroblasts. On the other hand, FA and TFA resulted in extensive depurination of nucleic acids while HFIP and TFE did so to a lesser degree. Among the investigated chemical scrapie inactivators, FA treatment could offer a safe and biocompatible collagen-derived material for biomedical use. PMID- 9358315 TI - Retention of enzymatic activity immobilized on silanized Co-Cr-Mo and Ti-6Al-4V. AB - Biochemical surface modification of biomaterials utilizes immobilized biomolecules to induce preferred tissue responses. Operational stability, or retention of biological activity, of biochemically modified biomaterials is a fundamental determinant of their usefulness. The present study investigated retention of enzymatic activity immobilized on silanized Co-Cr-Mo and Ti-6Al-4V. A model enzyme, trypsin, was immobilized on monolayers and multilayers of silane deposited from aqueous or organic solutions of gamma-aminopropyltriethoxysilane (APS). Trypsin-conjugated biomaterials were incubated in cell culture medium at 37 degrees C for up to 96 h, and the residual immobilized activity was measured. Retention of bioactivity in physiological saline was dependent on the type of material and on the method of silanization. Activity of enzyme adsorbed on the metals was lost within 24-48 h. Both mono- and multilayers of APS deposited on Co Cr-Mo by aqueous silanization effectively retained enzymatic activity throughout the 96 h incubation period. The monolayer retained approximately 23% of the activity initially present, and the multilayers retained approximately 50% of the initial activity. Organic silanization of Co-Cr-Mo was marginally effective as it initially slowed the loss of activity. However, all activity was lost by 48-72 h of incubation. Neither organic nor aqueous silanization enhanced retention of enzymatic activity on Ti-6Al-4V. PMID- 9358316 TI - Kinetics of blood component adsorption on poly(D,L-lactic acid) nanoparticles: evidence of complement C3 component involvement. AB - After intravenous administration, nanoparticles suffer a major drawback in that they are rapidly and massively taken up by the cells of the mononuclear phagocyte system. The mechanisms involved in the opsonization, adhesion, and internalization of biodegradable nanoparticles by the mononuclear phagocyte system are still poorly understood. In this work, the kinetics of blood protein adsorption onto nanoparticles of poly(D,L-lactic acid) prepared by the salting out technique was investigated. Nanoparticles of 312 nm were incubated for variable periods of time (5-60 min) in human serum and citrated plasma. After incubation, the particles were washed and the proteins detached from them, denatured, and analyzed by two-dimensional polyacrylamide gel electrophoresis. In plasma, the predominant protein was immunoglobulin G (IgG), and the amount adsorbed was not dependent on incubation time. Albumin amounts were high for short incubation periods but decreased as a function of time, whereas apolipoprotein E levels increased significantly as a function of the incubation period. Owing to the possible complement cascade inactivation by addition of citrate to plasma, the kinetics of adsorption was also evaluated in serum. In this medium, adsorption of complement C3 components onto the surface of the nanoparticles was clearly evidenced by spots of increasing intensity and area, reaching levels comparable to those of the omnipresent IgG. This result confirms the important role of complement components in the opsonization process of poly(D,L-lactic acid) particles. PMID- 9358318 TI - Crosslinking of hyaluronic acid with water-soluble carbodiimide. AB - Hyaluronic acid (HA) was chemically crosslinked with a water-soluble carbodiimide (WSC) to produce low-water-content films when brought into contact with water. The crosslinking reaction was performed in two different ways; one was by using HA films and the other by casting HA solutions. Both methods produced water insoluble HA films. The lowest water content of the crosslinked HA films subjected to swelling with water was 60 wt % at 37 degrees C, which was lower than any reported values. Infrared spectra of the crosslinked films suggested that intermolecular formation of ester bonds between the hydroxyl and carboxyl groups belonging to different polysaccharide molecules led to crosslinking. For comparison, pectin which possesses hydroxyl and carboxyl groups in one molecule, similar to HA, was subjected to crosslinking with WSC. The finding on pectin also supported ester formation between different polysaccharide molecules. The crosslinking of HA film with WSC in the presence of L-lysine methyl ester prolonged the in vivo degradation of HA film, probably because of amide bond formation as the crosslink. PMID- 9358317 TI - Long-term evaluation of bone-titanium interface in rat tibiae using light microscopy, transmission electron microscopy, and image processing. AB - We conducted a 2-year histologic and histometric evaluation of the tibial bone titanium (Ti) implant interface in male rats. Thirty male 6-week-old rats were used in this study. They were divided into two groups: 15 for day 28 and 15 for day 730. Microscopic observation at day 28 revealed that the newly formed bone around the implant almost surrounded the implant, but fibroblastlike cells were interposed in some histologic sections. At day 730, in contrast, such cells were rarely seen, and the bone around the implant presented a lamellar structure. Transmission electron microscopic observation at day 28 disclosed mature or poorly mineralized bone near the implant; however, an electron-dense amorphous zone about 50 nm in thickness was interposed between the bone and Ti. In places slender cells were interposed between the bone and Ti. The amorphous zone was also observed at the cell-Ti interface. At day 730, a poorly mineralized layer remained in some areas between the mature bone and the titanium, and the interposed amorphous zone was still observed. Occasionally, a 200-nm-thick layer, thought to be cell remnant, was seen. As calculated in an image-processing, system analysis, the percent bone contact and the thickness and area of the surrounding bone for the Ti implant at day 28 were 43.6%, 30.4 microns, and 0.10 mm2, respectively, and those at day 730 were 89.9%, 53.5 microns, and 0.19 mm2, respectively. In summary, although the passage of time may affect bone maturity, interfacial cells remain at the bone-Ti interface as a uniform layer together with unmineralized bone. PMID- 9358319 TI - A study on adsorption structures of methacryloyloxyalkyl dihydrogen phosphates on silver substrates by infrared reflection absorption spectroscopy. AB - 10-Methacryloyloxydecyl dihydrogen phosphate (M10P) for use in dentistry has recently been noted as an adhesive monomer contained in a metal primer. Although the treatment of a metal surface with primer before the application of resin is recognized to improve the adhesion between metal and resin, the role of M10P in the adhesion process has not been clarified. In this study, infrared reflection absorption (IRA) spectroscopy was employed to study the adsorption structures of M10P as well as 2-methacryloyloxyethyl dihydrogen phosphate (M2P) on evaporated silver substrates. The IRA spectra of the self-assembled films of those phosphates verified the adsorption of M10P or M2P on silver substrates from the methyl methacrylate solutions (5 x 10(-5) mol/L). The saturation coverages of M10P and M2P were completed after about 50 and 25 min, respectively. Two characteristic bands around 980 and 1080 cm-1 due to the PO(2-)3 stretching vibrations were observed. These results indicate that the phosphate groups of both monomers are adsorbed to silver surfaces in the dissociated form, -PO(2-)3, and form hydrophobic monolayers. The monolayer of M10P was found to be more durable against thermocycling in water than that of M2P by IRA measurements. The roles of M10P in the metal primer are presumably to form such a monolayer with appreciable durability and to promote polymerization with resin monomers. PMID- 9358320 TI - Effects of N-methacryloyl-omega-amino acid primer pretreatment on the bond strength of the resin to acid-etched dentin. AB - To ascertain the adhesion mechanism of resins to etched dentin treated with hydrophilic primers such as N-methacryloyl-omega-amino acids (NM omega A), the effect of the application of NM omega A primers on the bond strength of the resin and also the characteristics of the "hybrid layer" were investigated. Here, the concept of "hybrid layer" has been proposed previously by Nakabayashi et al. When the demineralized dentin was treated with the NM omega A solution, the bond strength increased remarkably, thus indicating the formation of a hybrid layer. It can be construed that NM omega A primers allowed for diffusion of the bonding agent to the dentinal collageous layer that was exposed by acid etching, and thereby the creation of a hybrid layer. To obtain an understanding of how NM omega A primers improved bond strength at the interface between the resin and dentinal collagen, the 13C-NMR spectra of NM alpha A were observed in the absence and presence of demineralized dentin. The 13C peak intensities of all of the carbons of the NM alpha A species were dramatically reduced in the presence of the dentin. Specifically, the reduction of the carbon peak intensity of carboxylic acid in the NM alpha A species was reduced by 30%. This indicated that the unionized carboxylic acid in the NM alpha A primer interacted with the dentinal collagen. Thus, the composite resin can be considered to adhere to the dentinal collagen through the unionized NM alpha A that interacts with the dentinal collagen. PMID- 9358321 TI - Bonding of chemically treated titanium implants to bone. AB - A study was undertaken in rabbit tibiae to determine the effects of chemical treatments and/or surface-induced bonelike apatite on the bone-bonding ability of titanium (Ti) implants. Smooth-surfaced plates (10 x 10 x 2 mm) of pure Ti, alkalil- and heat-treated Ti, and bonelike apatite-formed Ti after the treatments were implanted into the tibial metaphyses of mature rabbits. The tibiae containing the implants were harvested at 4, 8, and 16 weeks after implantation and subjected to a tensile testing and histologic evaluation. Biomechanical results showed that both treated implants exhibited significantly higher failure loads compared with untreated Ti implants at all time periods. Histologic examination by Giemsa surface staining, contact microradiography (CMR), and scanning electron microscopy (SEM) in backscatter mode revealed that both treated Ti implants directly bonded to bone tissue during the early postimplantation period, whereas untreated Ti implants formed direct contact with the bone only at 16 weeks. SEM-electron-probe microanalysis (EPMA) examination showed a Ca-P-rich layer at the interface between the treated implants and bone, although the Ca-P rich layer was not detected on the surface of untreated implants during observation periods. The results of this study suggest that chemical treatments may accelerate the bone-bonding behavior of titanium implants and enhance the strength of bone-implant bonding by inducing a bioactive surface layer on Ti implants. PMID- 9358322 TI - Effect of gravity and diffusion interface proximity on the morphology of collagen gels. AB - Collagen solutions (0.25% w/v) were polymerized in microgravity (STS-77, 10 days) along with simultaneous ground controls. Assembly conditions were achieved by the passage of buffer ions across a dialysis membrane into a reaction chamber containing the dissolved collagen. The gels were analyzed macroscopically and microscopically to assess the influence of gravity and the oriented diffusion of buffer ions on the resulting product. Double-blind rankings based on visual observation of the gels established that all of the flight gels (n = 8) were more uniform in appearance than all of the ground gels (n = 6). Photography using side illumination of the gels revealed the more granular appearance of the ground gels relative to the highly uniform appearance of the flight gels. Scanning electron microscopy established this difference at the microscopic level. Proximity to the dialysis interface and the presence or absence of gravity were both found to control the porosity and uniformity of the matrix. PMID- 9358324 TI - The effect of heat treatment on the fatigue strength of microknurled Ti-6Al-4V. AB - Microknurling, a high pressure surface indentation technique, was devised as an alternative to traditional heat-bonded porous coatings found on many orthopedic implants designed for fixation by tissue ingrowth. Heat-bonded porous coating can cause at the surface of an implant stress concentrations that reduce fatigue strength. However, microknurling may reduce stress intensification without eliminating it. Thus the purpose of this work was to explore surface thermal/mechanical processing of Ti-6Al-4V to improve the fatigue strength of microknurled specimens via the production of a Ti-6Al-4V dual microstructure. The latter consists of a surface layer of equiaxed grains known to be effective against crack initiation and a bulk microstructure of lamellar grains that possesses optimum fatigue crack propagation resistance. Rotating-bending fatigue tests showed that such a microstructure had some benefits, but this was offset by the reduction in compressive strains imparted to the surface by the heat treatments needed to obtain this microstructure. PMID- 9358325 TI - An example of using mixed models and PROC MIXED for longitudinal data. AB - Longitudinal data, or data that are repeated measurements on various subjects across time, are commonplace in biostatistical studies. The general linear mixed model is a useful statistical tool for analyzing such data and drawing meaningful inferences about them. This paper discusses some of the most common mixed models and fits them to a prototypical example involving repeated measures on blood pressure. Computer implementation is via the MIXED procedure in the SAS System, and code descriptions and output interpretations accompany the example. PMID- 9358323 TI - A photochemical method for the surface modification of poly(etherurethanes) with phosphorylcholine-containing compounds to improve hemocompatibility. AB - Phosphorylcholine groups attached to polymer surfaces are known to improve hemocompatibility. A photochemical method is presented to couple phosphorylcholine-containing aryl azides to poly(etherurethane) surfaces (PEUs). Two aryl azides that consist of a photoactivatable 4-azidobenzoyl group, a short spacer chain, and a phosphorylcholine endgroup were synthesized. The two compounds differ only in the type of spacer used: triethylene glycol for compound 1 and hexanediol for compound 2. These compounds were physically adsorbed to PEU surfaces. Upon UV irradiation, reactive intermediates are formed that react with nucleophilic groups on the polymer surface. The modified surfaces showed decreased underwater contact angles, indicating that hydrophilic phosphorylcholine groups are present at the surface. ESCA measurements showed the presence of phosphorus and positively charged nitrogen atoms in the outermost polymer layers (analyzed depth about 50 A), which is a strong indication of the presence of phosphorylcholine groups. Hemocompatibility in vitro was tested with thrombin generation assays and platelet adhesion tests. In thrombin generation assays the clotting time of platelet-rich plasma in contact with the polymer surface is determined. Clotting times were clearly prolonged for the modified surfaces. Surfaces modified with compound 2 showed slightly higher clotting times than those modified with compound 1. Repeated surface modification with compound 2 further increased the clotting time. For the tested surfaces an increase in the clotting time corresponds to an increase in the concentration of phosphorylcholine groups at the surface (as measured by ESCA and contact angle). Platelet adhesion studies with scanning electron microscopy demonstrated that fewer platelets (showing less activation) adhered to the modified surfaces than to the unmodified polyurethane. PMID- 9358327 TI - Interim analyses and early termination of clinical trials. AB - The group sequential tests (GST) are appropriate for performing interim analyses in clinical trials. Various GST are reviewed and compared in this paper in terms of the expected sample size, the maximum sample size, and other practical aspects. Also discussed are the p-values of the significant differences for GST. Common problems and difficulties of using GST in practice are examined. One problem is difficulties associated with the delayed data accumulated after a trial is terminated at an interim test. The GST with O'Brien-Fleming type of boundaries are found to be safe in dealing with delayed observations. Possible approaches for performing futility analyses are illustrated with examples. It is recommended that futility analysis with GST be built into the design of large clinical trials. PMID- 9358326 TI - Longitudinal analyses for magnetic resonance imaging outcomes in multiple sclerosis clinical trials. AB - A basic feature of many clinical trials is the collection of longitudinal data on individual patients. Analysis of such data is often based on summaries over time. This allows use of standards methods to assess treatment effects but sacrifices information on patterns over time as well as potential greater efficiency of analysis. The purpose of this paper is to illustrate the utility of the generalized estimating equations (GEE) approach to the analysis of longitudinal binary, count, and continuous responses for the frequent magnetic resonance imaging (MRI) substudy of the 3-year pivotal trial of interferon beta-1 b in relapsing-remitting multiple sclerosis. PMID- 9358328 TI - A reviewer's perspective on multiple endpoint issues in clinical trials. AB - Multiplicity issues due to clinical endpoints frequently arise in clinical trials. Conducting tests of significance separately for each endpoint in a univariate manner, or ignoring the issue, could lead to inflation of the type 1 error probability in making treatment effect claims. This is of concern because inflation of the type I error probability could lead to approval of inefficacious therapies. Therefore, one generally requires that this error probability be controlled at some prespecified alpha-level. At the same time the method employed for this purpose should be one with optimal efficiency so as to be able to detect clinically meaningful treatment effect with high probability. In this presentation, we give a clinical and statistical background to the problem with a few examples and show some simulation results that illustrate the impact of ignoring multiplicity due to multiple endpoints on the type I error probability. This is then followed by an overview and discussion of some global methods in the literature and how they can be used to make endpoint specific tests of significance. Finally, we will introduce a Monte-Carlo simulation and resampling approach (with examples using real data) for controlling the type I error probability. PMID- 9358329 TI - In vitro-in vivo relationships for oral extended-release drug products. AB - It is of interest to predict the in vivo behavior of an oral extended-release drug product based on its in vitro dissolution profile. In some cases a suitable convolution-based prediction model can be found. We present a methodology for developing statistical models of in vitro-in vivo relationships under the framework of the mixed-effects nonlinear model and discuss methods for assessing the validity and strength of the relationship. These methods are illustrated and contrasted with a level A in vitro-in vivo correlation using data from a study involving four different formulations of an oral extended-release drug product. PMID- 9358330 TI - Statistical applications for in vitro diagnostic tests and other medical device clinical trials. AB - Some statistical methods applied to in vitro diagnostic tests for the three primary indications (screening, diagnosis, and monitoring) are discussed. Various examples with practical statistical applications are presented, including test for k by k ordered categorical matched-pair data for screening of cervical cancer, receiver operating characteristic (ROC) curve for diagnosis or screening, and the Cox time-to-event model to estimate relative risk of first cancer progression by monitoring carcinoembryonic (CEA) levels for stage IV breast cancer patients. PMID- 9358331 TI - Analysis and design of repeated measures in clinical trials using summary statistics. PMID- 9358333 TI - Adaptive survival trials. AB - We present a design for adaptive survival trials, where the probability of randomization to one of two treatments is skewed away from 0.5 according to the current value of the logrank statistic. A formula mapping the logrank statistic onto [0,1] is given, which is then used to bias a coin used for randomization. Simulation evidence shows that the allocation scheme works well and offers a more ethical alternative when lifetime data are available from other patients during the recruitment period. Power is not adversely affected by the resulting unequal allocation. The usual test statistic appears to be standard normal under the proposed allocation scheme. PMID- 9358334 TI - Statistical strategies for event rate comparisons in dental studies. AB - Dental researchers are interested in measures of association between explanatory variables and various disease indicators, where both can be measured at the tooth or surface level. The usual chi-square tests for event rate comparisons are inadequate to analyze such data since teeth within the same individual are correlated with one another. This situation may become further complicated when stratification adjustment is of interest, particularly when the stratification factor is also at the tooth or surface level, such as tooth type or presence of previous disease for the tooth. Consequently, statistical methods are needed to control for the correlation of sites in the mouth. A survey sampling approach is suggested; with this method, the sample is managed as a one stage cluster sample of patients to obtain the design base covariance matrix for the cell counts in a contingency table. The covariance matrix for the cell counts can be obtained from statistical software used to analyze data from large sample surveys. Approximate estimates for variance for various measures of association can then be obtained by Taylor series methods. Event rate comparisons through the odds ratio, relative risk, risk difference as well as other measures of association can be made with adjustments for the intra-patient correlation. Methods for summary measures of association across strata are considered as well. PMID- 9358332 TI - Evolution of the CANDA at Roche. AB - Computer-assisted NDAs (CANDAs) have evolved from an instrument that provides information to regulators to also providing information to internal researchers. They are still not as useful internally during the NDA process as they will be in the future. The portion of CANDA that allows summary, display, and query of the raw data is the most complicated part of the CANDA. Roche's current CANDA system, as well as the challenges in using, documenting, and validating such a system, is described, stressing the features that make it usable in-house. Roche's vision of the future is outlined, covering the strategies, targets, and expected savings. PMID- 9358335 TI - A logistic dose-ranging method for phase I clinical investigations trials. AB - This paper describes an alternative to the continual reassessment method (CRM) for phase I trials. The logistic dose ranging strategy (LDRS) uses logistic regression and a dose allocation scheme similar to the CRM. It can easily be implemented from any logistic regression program. The LDRS can be a stand alone dose allocation scheme or it can be incorporated into standard three on a dose strategies to indicate when escalation can proceed more rapidly. Finally, the effect of covariates such as age or comorbid conditions on the toxicity expected for the dose selected for a phase II trial can be examined. PMID- 9358336 TI - Crossover trials with a cumulative response. AB - We consider the use of a crossover design when the response is long-lasting and cumulative. Examples include the treatment of chronic diseases such as cerebral palsy, and educational and psychological intervention studies. Cumulative response models are developed for the two-treatment, two-period design. Various estimators for the treatment and carryover effects are compared, and ANOVA tables are derived. The crossover design including pretreatment observations is found to be more cost-efficient than the one-period randomised design. Without pretreatment data, it may be more or less efficient, depending on the reliability of the data, and the costs of subject recruitment, treatment, and observation. PMID- 9358337 TI - Sensitive monoclonal antibody-based sandwich ELISA for determination of the diabetes-associated autoantigen glutamic acid decarboxylase GAD65. AB - Although various methods for the detection of autoantibodies against glutamic acid decarboxylase (GAD65-AAb) are known, no sensitive method for the quantification of GAD65 as autoantigen is available. We describe a sandwich ELISA based on monoclonal GAD65 antibodies (Mc-GAD65-Ab) of different epitope specificities to quantify GAD65 in pancreatic islets and in different organ/cell extracts and during the preparation of GAD from brain extracts. GAD65 was captured via solid phase coated Mc-GAD65-Ab and detected via a second biotin labelled Mc-GAD65-Ab recognizing a NH2-terminal epitope of the molecule. The detection limit was estimated to be 0.03 ng GAD65/ml using alkaline phosphatase (AP)-conjugated streptavidin. GAD65 contents in islets of neonatal BB/OK rats and Lewis rats amounted to 37.4 and 43.7 pg/islet, respectively. Furthermore, GAD65 was quantified in brain extracts of pig (55.1 ng/mg protein), mouse (39.5 ng/mg), rat (243.8 ng/mg) and pig cerebellum (514.8 ng/mg) and in different organ extracts of Lewis rat. PMID- 9358339 TI - A simple and rapid immunoassay system using green fluorescent protein tag. AB - A fusion protein between green fluorescent protein (GFP) and neuron-specific enolase (NSE) was expressed in Escherichia coli. The GFP-NSE fusion protein migrated at 62 kDa in SDS-PAGE and retained the fluorescence under non-heating conditions. However, heat-denatured GFP-NSE was non-fluorescent and migrated at 74 kDa corresponding to the theoretical value. This suggests that the special structure of GFP, which is not denatured by SDS, influences its mobility in SDS PAGE under non-heating conditions. The fluorescence intensity of GFP-NSE was measurable over a wide range by spectrophotometry or densitometry. The competitive immunoassay for NSE was performed using GFP-NSE as labeled antigen. Under our assay conditions, the working range of this system was about 2 -60 ng. This simple and rapid fluorescence immunoassay (FIA) using GFP-tagged antigen may be applicable to many protein markers. PMID- 9358338 TI - Characterization of monoclonal antibody reactive with 10-hydroxy-12(Z) octadecenoic acid (10-OHODA) and its demonstration in cultured human macrophages. AB - 10-Hydroxy-12(Z)-octadecenoic acid (10-OHODA) has an inhibitory effect on the tension of papillary muscles in isolated guinea-pig hearts. To establish an immunoassay for 10-OHODA a mouse monoclonal antibody (MoAb), YM-1, was produced. In order to evaluate the ability of this MoAb to recognize various 10-OHODA analogs including leukotoxin (9, 10-epoxy-12-octadecenoic acid, LTx), a sensitive enzyme-linked immunosorbent assay (ELISA) was developed using the avidin-biotin complex (ABC). The detection limit for 10-OHODA was as low as 0.5 ng in this system. In order to demonstrate the presence of 10-OHODA in living cells, macrophages derived from the human leukemia cell line THP-1 by adding 160nM phorbol 12-myristate 13-acetate (PMA) were exposed to 95% O2, and 5% CO2 for 24 h. 10-OHODA and other fatty acids were extracted from the exposed macrophages with diethylether after phospholipase A2 treatment. The 10-OHODA content was determined using the new ELISA, and 18.5 ng 10-OHODA was detected in the macrophages exposed to the high oxygen concentration (1 x 10(6) cells). PMID- 9358340 TI - An enzyme immunoassay for rat growth hormone: validation and application to the determination of plasma levels and in vitro release. AB - A competitive enzyme immunoassay for rat growth hormone (rGH) has been developed using polyclonal anti-rGH antibodies and an acetylcholinesterase (EC 3.1.1.7.) enzymatic tracer coupled covalently with rGH. The assay was performed in 96-well microtiter plates coated with rabbit polyclonal anti-goat immunoglobulin antibodies. Molecular sieve filtration and Western blot analysis revealed a single immunoreactive peak for rat plasma or pituitary extracts. Cross-reactivity with other rat pituitary hormones or human GH was less than 1%. Assay of samples in a concentration range of 0.7 to 69 ng/ml by enzyme immunoassay and radioimmunoassay were well correlated (r = 0.87 and 0.85 respectively for plasma and culture medium samples). Intra- and inter-assay variations in plasma were 4 (n = 24) and 14% (n = 9) respectively. Minimal detectable amounts of rGH were 0.6 ng/ml. A two-site immunometric assay also developed with the same antibodies allowed a detection threshold of 0.25 ng/ml. PMID- 9358341 TI - Production of monoclonal antibodies against Nosema bombycis and their utility for detection of pebrine infection in Bombyx mori L. AB - Latex agglutination assay based on monoclonal antibodies (MCAs) described in this communication may be useful for detection of Pebrine infection in silkworm. Four murine MCAs were produced against Nosema bombycis spore. In ELISA all 4 MCAs (IgM isotype) reacted with alkali treated Nosema spores and to variable extent with acetone precipitated surface protein. However, MA-310 and MA-542 showed a low degree of cross reactivity with BmNPV. In contrast, MA-503 and MA-515 were devoid of reactivity with BmNPV, B. thuringiensis, S. marcescens, Azotobactor, Rhizobium and normal hemolymph protein in ELISA. Latex beads sensitized with a combination of MA-503 and MA-515 (50 micrograms each per ml of 0.4% latex beads) could detect 1 x 10(5) Nosema spores per test. Sensitization of the latex beads with the cocktail of these two MCAs through protein-A bridge further led to a 10-fold increase in the sensitivity (1 x 10(4) spores/test) of the assay. No agglutination was observed in presence of BmNPV, Rhizobium, Azotobactor, E. coli, B. thuringiensis, S. marcescens and normal hemolymph protein indicating the specificity of the test. The results obtained by latex agglutination assay on hemolymph samples of infected as well as normal larvae collected from field, II instar larvae infected in the laboratory and from infected mother moth revealed 100% correlation with results by microscopic examination. PMID- 9358342 TI - Enhanced cytokine detection by a novel cell culture-based ELISA. AB - Production of some cytokines, such as IL-4 and IL-10, often occurs at low levels and is difficult to detect by standard ELISA techniques. In many cases the level of detection is at or near to the limits of sensitivity of the assay due either to minimal synthesis and/or cytokine consumption. In an effort to enhance the quantitation of weakly detected cytokines we have developed a unique cell culture capture ELISA. Lymphocytes are incubated in an anti-cytokine antibody coated ELISA plate for the last 6 hours of a 24 hour in vitro activation period. Use of this cell culture capture method consistently enhanced detection of several T cell cytokines compared to conventional ELISA techniques. Moreover, this technique was found to enhance detection without altering the rate of cytokine secretion which occurred prior to the cell culture capture period. Thus, the cell culture capture ELISA may be useful for detection of a variety of cytokines which are produced at low levels and have traditionally been difficult to quantify. PMID- 9358343 TI - Aging, research and families. PMID- 9358344 TI - Corporate tyranny. PMID- 9358346 TI - The medical student and the suicidal patient. AB - Today's medical students are being confronted with ethical situations of far greater complexity than were their predecessors and yet the medical education system does little to prepare students for the ethical dilemmas which they inevitably face when entering the hospital environment. The following article addresses the issues surrounding a case where a patient has told a student in confidence of his plans to commit suicide. What should the student do? The only way for the student to prevent death is by breaking confidentiality because the student has insufficient clinical experience to provide adequate guidance. However, this requires ignoring the patient's right to autonomy, a right enshrined in both case law and medical ethics. Clearly the student's ethical, moral and legal position must be carefully evaluated. PMID- 9358345 TI - Experienced consent in geriatrics research: a new method to optimize the capacity to consent in frail elderly subjects. AB - OBJECTIVES: Cognitive and sensory difficulties frequently jeopardize informed consent of frail elderly patients This study is the first to test whether preliminary research experience could enhance geriatric patients' capacity to consent. DESIGN/SETTING: A step-wise consent procedure was introduced in a study on fluid balance in geriatric patients. Eligible patients providing verbal consent participated in a try-out of a week, during which bioelectrical impedance and weight measurements were performed daily. Afterwards, written informed consent was requested. Comprehension, risk and inconvenience scores (ranges: 0 10) were obtained before and after the try-out by asking ten questions about the study's essentials and by asking for a risk and inconvenience assessment on a ten points rating scale. SUBJECTS AND RESULTS: Seventy of the 78 eligible subjects started the try-out and 53 (68%) provided written consent. The comprehension score increased from 5.0 (+/- 2.3) to 7.0 (+/- 1.9) following the try-out (P < 0.001). The number of subjects capable of weighing risks and inconveniences increased from 32 to 48 (P < 0.001). CONCLUSIONS: Research experience improved the capacity to consent, still enabling an acceptable participation rate. Therefore, experienced consent seems a promising tool to optimize informed consent in frail elderly subjects. PMID- 9358347 TI - Should informed consent be based on rational beliefs? AB - Our aim is to expand the regulative ideal governing consent. We argue that consent should not only be informed but also based on rational beliefs. We argue that holding true beliefs promotes autonomy. Information is important insofar as it helps a person to hold the relevant true beliefs. But in order to hold the relevant true beliefs, competent people must also think rationally. Insofar as information is important, rational deliberation is important. Just as physicians should aim to provide relevant information regarding the medical procedures prior to patients consenting to have those procedures, they should also assist patients to think more rationally. We distinguish between rational choice/action and rational belief. While autonomous choice need not necessarily be rational, it should be based on rational belief. The implication for the doctrine of informed consent and the practice of medicine is that, if physicians are to respect patient autonomy and help patients to choose and act more rationally, not only must they provide information, but they should care more about the theoretical rationality of their patients. They should not abandon their patients to irrationality. They should help their patients to deliberate more effectively and to care more about thinking rationally. We illustrate these arguments in the context of Jehovah's Witnesses refusing life-saving blood transfusions. Insofar as Jehovah's Witnesses should be informed of the consequences of their actions, they should also deliberate rationally about these consequences. PMID- 9358348 TI - May we practise endotracheal intubation on the newly dead? AB - Endotracheal intubation (ETI) is a valuable procedure which must be learnt and practised, and performing ETI on cadavers is probably the best way to do this, although lesser alternatives do exist. Performing ETI on a cadaver is viewed with a real and reasonable repugnance and if it is done without proper authorisation it might be illegal. Some form of consent is required. Presumed consent would preferably be governed by statute and should only occur if the community is well informed and therefore in a position of being able to decline. Currently neither statute nor adequate informing exists. Endotracheal intubation on the newly dead may be justifiable according to a Guttman scale if the patient has already consented to organ donation and if further research supports the relevance of the Guttman scale to this question. A "mandated choice" with prior individual consent as a matter of public policy is the best of these solutions, however until such a solution is in place we may not practise endotracheal intubation on the newly dead. PMID- 9358349 TI - Doctors' stories, patients' stories: a narrative approach to teaching medical ethics. AB - Many senior doctors have had little in the way of formal ethics training, but express considerable interest in extending their education in this area. This paper is the report of an initiative in continuing medical education in which doctors were introduced to narrative ethics. We review the theoretical basis of narrative ethics, and the structure of and response to the two-day workshop. PMID- 9358350 TI - Ethical issues in long-term psychiatric management. AB - Two general ethical problems in psychiatry are thrown into sharp relief by long term care. This article discusses each in turn, in the context of two anonymised case studies from actual clinical practice. First, previous mental health legislation soothed doubts about patients' refusal of consent by incorporating time limits on involuntary treatment. When these are absent, as in the provisions for long term care which have recently come into force, the justification for compulsory treatment and supervision becomes more obviously problematic. Second, Anglo-American law does not normally allow the preventive detention of someone who may be dangerous but has not actually committed any crime. The justification for detaining a possibly dangerous user of mental health services without his or her consent can only be based on risk assessment, but this raises issues of moral luck. Is the psychiatrist who decides not to take out a supervision order for a possibly dangerous patient with an initial psychotic diagnosis morally at fault if that person harms someone in the community, or himself? Or is the psychiatrist merely unlucky? PMID- 9358351 TI - Moral assessment of growth hormone therapy for children with idiopathic short stature. AB - The prescription of growth hormone therapy for children who are not growth hormone deficient is one of the controversies in contemporary paediatric endocrinology. Is it morally appropriate to enhance the growth, by means of medical treatment, of a child wish idiopathic short stature? The medical, moral, and philosophical questions in this area are many. Data on the effects of human growth hormone (hGH) treatment will not on their own provide us with answers, as these effects have to be evaluated from a normative perspective. In this article we consider hGH treatment for children of idiopathic short stature from three normative perspectives: the goals of medicine, the good of the patient, and the public good. We argue that the prevention of psychological and social problems due to short stature (and not merely the enhancement of growth) should be the ultimate goal of medical treatment and research. PMID- 9358352 TI - Health care, human worth and the limits of the particular. AB - An ethics concerned with health care developments and systems must be historically continuous, especially as it concerns the application to managed structures of key moral-epistemic concepts such as care, love and empathy. These concepts are traditionally most at home in the personal, individual domain. Human beings have non-instrumental worth just because they are human beings and not by virtue of their capacities. Managed health care systems tend to abstract from this worth in respect of both individuals' distinctness and individual identity. The first, a common feature of quantitative approaches to health care assessment and delivery, is avoidable. The second, by contrast, is necessarily sacrificed in impersonally managed structures. Failure to distinguish the two encourages confusion and distress, and the demand for impossible medico-moral relationships. PMID- 9358353 TI - Teaching ethics using small-group, problem-based learning. AB - Ethics is the emphasis of our first-year Introduction to Clinical Medicine-1 course. Introduction to Clinical Medicine-1 uses problem-based learning to involve groups of seven to nine students and two facilitators in realistic clinical cases. The cases emphasize ethics, but also include human behaviour, basic science, clinical medicine, and prevention learning issues. Three cases use written vignettes, while the other three cases feature standardized patients. Groups meet twice for each case. In session one, students read the case introduction, obtain data from the written case or standardized patient, identify the case's ethical problems, formulate learning issues, discuss ways to resolve the moral conflicts, and assign research responsibilities. In session two, students discuss their assigned learning issues and specify and justify clinical actions to address the case's ethical dilemmas. Following three cases, groups write an essay discussing what they learned and describing how they would approach and resolve the case's learning issues. PMID- 9358354 TI - Medical negligence and wrongful birth actions: Australian developments. AB - Wrongful birth actions aim to compensate litigants who are negligently deprived by health professionals of their right to reproductive choice. Access to safe and legal abortion is integral to the action and wrongful birth claims in the United Kingdom have been facilitated by the Abortion Act 1967 (as amended). The recent Australian case CES v Superclinics (1995) 38 NSWLR 47 shows how judicial confusion about the legality of abortion can result in judges condoning medical negligence. The Superclinics case also suggests that doctors are not required to provide pregnant women with the same standard of care as other patients. These developments show that law can become incoherent and health professionals can act negligently with impunity when reproductive choice does not have a secure legal foundation. PMID- 9358356 TI - Compensation for the subjects of medical research. PMID- 9358355 TI - Medical decisions concerning the end of life: a discussion with Japanese physicians. AB - OBJECTIVES: Life-sustaining treatment at the end of life gives rise to many ethical problems in Japan. Recent surveys of Japanese physicians suggested that they tend to treat terminally ill patients aggressively. We studied why Japanese physicians were reluctant to withhold or withdraw life-support from terminally ill patients and what affected their decisions. DESIGN AND PARTICIPANTS: A qualitative study design was employed, using a focus group interview with seven physicians, to gain an in-depth understanding of attitudes and rationales in Japan regarding medical care at the end of life. RESULTS: Analysis revealed that physicians and patients' family members usually make decisions about life sustaining treatment, while the patients' wishes are unavailable or not taken into account. Both physicians and family members tend to consider withholding or withdrawing life-sustaining treatment as abandonment or even killing. The strongest reason to start cardiopulmonary resuscitation- and to continue it until patients' family members arrive-seems to be the family members' desire to be at the bedside at the time of death. All physicians participating in our study regarded advance directives that provide information as to patients' wishes about life-sustaining treatment desirable. All expressed concern, however, that it would be difficult to forego or discontinue life-support based on a patient's advance directive, particularly when the patient's family opposed the directive. CONCLUSION: Our group interview suggested several possible barriers to death with dignity and the appropriate use of advance directives in Japan. Further qualitative and quantitative research in this regard is needed. PMID- 9358357 TI - Cost effectiveness of medical ethics training. PMID- 9358358 TI - The ways and wherefores of immediate placement of implants into fresh extraction sites: a literature review. AB - A waiting period of 12 months or longer to allow total socket healing used to be accepted protocol for placing dental implants. More than 15 years of research and clinical practice were needed for the concept of immediate endossceous implantation into fresh extraction sites to be accepted. Today the dilemma is no longer when, but which, protocol to follow. The diverse recommendations found in the literature leave the practitioner confused as to the methodology of choice. The conclusions drawn after reviewing the relevant literature on immediate dental implantation are: 1) implants placed into fresh extraction sockets have a high rate of survival, ranging between 93.9% to 100%; 2) implants must be placed 3 to 5 mm beyond the apex in order to gain a maximal degree of stability; 3) implants should be placed as close as possible to the alveolar crest level (0 to 3 mm); 4) there is no consensus regarding the need for gap filling and the best grafting material; 5) the use of membrane does not imply better results-on the contrary, membrane exposure may carry complications in its wake; and 6) the absolute need for primary closure remains to be established. PMID- 9358359 TI - Histological comparison of early wound healing following dense hydroxyapatite granule grafting and barrier placement in surgically-created bone defects neighboring implants. AB - The purpose of this study was to examine early wound healing following grafting of dense hydroxyapatite granules (HA granules) and barrier placement in surgically-created bone defects surrounding implants. Eight healthy adult dogs with an average weight of 15 kg were used in this study. Thirty-two bone defects measuring 4 mm x 4 mm were removed with a surgical bur to form continuous bucco lingual bone defects and 32 implants (16 titanium [Ti]) and 16 hydroxyapatite coated [HA]) were then placed into the defects. Four implant groups were created: 1) grafting HA; 2) covering with an expanded polytetrafluoroethylene (ePTFE) membrane; 3) grafting HA and covering with ePTFE membrane; and 4) control (no treatment). Animals were sacrificed 28 days after surgery. Histological sections revealed large amounts of newly-formed bone in all bone defects surrounding the implants treated with ePTFE membranes alone. Fibrous encapsulation of HA granules was observed in the defects of the HA granules grafting group. In the group with grafting of HA granules and covering with ePTFE membranes, small amounts of bone tissue were observed among HA granules, but most HA granules were surrounded with fibrous tissue. Bone defects were completely filled with connective tissue in the control group. There were no differences in the histological findings between Ti and HA-coated implants in all cases. Histomorphometric data disclosed that the presence of HA granules in the bone defects significantly arrested bone formation. Our study suggests that the grafting of dense HA into bone defects surrounding implants will result in fibrous healing during the early healing stage. PMID- 9358360 TI - Effects of guided bone regeneration around commercially pure titanium and hydroxyapatite-coated dental implants. II. Histologic analysis. AB - The purpose of this study was to determine which treatment of a large osseous defect adjacent to an endosseous dental implant would produce the greatest regeneration of bone and degree of osseointegration: barrier membrane therapy plus demineralized freeze-dried bone allograft (DFDBA), membrane therapy alone, or no treatment. The current study histologically assessed changes in bone within the healed peri-implant osseous defect. In a split-mouth design, 6 implants were placed in edentulous mandibular ridges of 10 mongrel dogs after preparation of 6 cylindrical mid-crestal defects, 5 mm in depth, and 9.525 mm in diameter. An implant site was then prepared in the center of each defect to a depth of 5 mm beyond the apical extent of the defect. One mandibular quadrant received three commercially pure titanium (Ti) screw implants (3.75 x 10 mm), while the contralateral side received three hydroxyapatite (HA) coated root-form implants (3.3 x 10 mm). Consequently, the coronal 5 mm of each implant was surrounded by a circumferential defect approximately 3 mm wide and 5 mm deep. The three dental implants in each quadrant received either DFDBA (canine source) and an expanded polytetrafluoroethylene membrane (ePTFE), ePTFE membrane alone, or no treatment which served as the control. Clinically, the greatest increase in ridge height and width was seen with DFDBA/ePTFE. Histologically, statistically significant differences in defect osseointegration were seen between treatment groups (P < 0.0001: DFDBA/ePTFE > ePTFE alone > control). HA-coated implants had significantly greater osseointegration within the defect than Ti implants (P < 0.0001). Average trabeculation of newly formed bone in the defect after healing was significantly greater for HA-coated implants than for titanium (P < 0.0001), while the effect on trabeculation between treatments was not significantly different (P = 0.14). Finally, there were significantly less residual allograft particles in defect areas adjacent to HA-coated implants than Ti implants (P = 0.0355). The use of HA-coated implants in large size defects with DFDBA and ePTFE membranes produced significantly more osseointegration histologically than other treatment options and more than Ti implants with the same treatment combinations. The results of this study indicate that, although the implants appeared osseointegrated clinically after 4 months of healing, histologic data suggest that selection of both the implant type and the treatment modality is important in obtaining optimum osseointegration in large size defects. PMID- 9358362 TI - Tooth loss due to periodontal abscess: a retrospective study. AB - This retrospective study focused on the frequency of tooth loss due to periodontal abscess among 42 patients who were treated by a single clinician over a 5- to 29-year period. A total of 114 patients were selected from the active periodontal recall schedule of a single periodontist at The University of Iowa College of Dentistry. The criteria for inclusion in the study included having a history of moderate to advanced periodontitis, being on 3 to 6 month recall periodontal maintenance care, and completion of active periodontal therapy prior to October 1987. Other parameters evaluated were age; gender; number of teeth present and missing at the initial, reevaluation, and last periodontal recall visit; initial periodontal prognosis; furcation involvement; non-surgical and surgical periodontal therapy; and reasons for tooth loss. Patients were grouped according to the number of teeth lost following active periodontal treatment into well-maintained (0 to 3), downhill (4 to 9), and extreme downhill (10 to 23) groups. Forty-two of the 114 patients were identified as having one or more periodontal abscesses. A total of 109 teeth were affected by periodontal abscess of which 49 (45%) teeth were lost and 60 (55%) were successfully maintained over an average of 12.5 years (5 to 29 years). More furcated teeth were lost than nonfurcated teeth and teeth given a hopeless prognosis were lost more consistently than those given a questionable prognosis in all groups. The frequency of periodontal abscess and tooth loss per patient was greater in the downhill and extreme downhill response groups than the well-maintained group. This suggests that teeth with a history of periodontal abscess can be treated and maintained for several years. PMID- 9358361 TI - Quantitative measurement of volume changes induced by oral plastic surgery: validation of an optical method using different geometrically-formed specimens. AB - The purpose of this research was to study the validity and variability of a projection Moire system, measuring volume differences of geometrically different formed specimens mimicking localized alveolar ridge defects. Nine pairs of specimens were fabricated, each of which simulated a preoperative ridge defect and a corresponding surgically-corrected postoperative ridge defect. All specimen pairs had a mathematically defined form which allowed the accurate assessment of their volume differences by a mechanical 3-D coordinate measuring machine or by a software-controlled milling machine. Measurements achieved with these methods were used as the references for comparison. Six specimen pairs, A1 to A6, possessed a simple rectangular geometrical form which facilitated their fabrication. Three specimen pairs, B1 to B3, were milled and consisted of geometrically more complex 3-D sculptured surfaces, which came closest to a true imitation of a localized ridge defect. An optical measurement system in the form of the projection Moire was utilized, applying a 4-phase shift technique, and results obtained with this device were regarded as test volumes. The absolute variability of the test volume measurements differed between 0.397 mm3 to 15.872 mm3, corresponding to a relative variability of 0.83% to 2.83%. The mean of the relative variability was within 1.68% for the "A" specimens and 2.15% for the "B" specimens. However, the difference was not significant, probably due to the limited number of "B" specimens. The systematic error of the Moire measurements in relation to the reference methods was surprisingly low, ranging from -0.12 mm3 to 7.67 mm3. The relative systematic error, expressed as a percentage of reference volume, ranged between 0.06% and -2.23%. The mean of the relative error for the more complex "B" specimens was 1.37%, which was less accurate in comparison to the more simply formed "A" specimens with a relative systematic error of 0.35%. Therefore, in this in vitro model it was possible to measure volume differences of geometrically different formed specimens, mimicking localized alveolar ridge defects, with a validity within 2.2% and with a variability of less than 2.8%. PMID- 9358363 TI - Pathologic migration of anterior teeth in patients with moderate to severe periodontitis. AB - The purpose of this study was to determine the prevalence of pathologic migration of anterior teeth in patients with moderate to severe periodontitis. The correlation between pathologic migration of anterior teeth and attachment loss (AL) was investigated, and an attempt was made to identify the most common early form of pathologic migration. Prevalence of tooth migration was studied in a group of 343 patients with moderate to severe periodontitis before treatment. The presence of pathologic migration was determined from the chief complaint and patient awareness o tooth movement in the last 5 years. Forty-four patients (age range 18 to 69; mean = 48.75) with 75 pairs of migrated and non-migrated teeth were studied further to determine if there is a correlation between severity of periodontal AL and pathologic migration. Migrated teeth were compared to control contralateral teeth that did not have migration. In addition, tooth mobility of the anterior teeth on 36 of the 44 patients was measured using the mobility meter. It was anticipated that tooth mobility would follow the same pattern as AL in relation to pathologic migration. The type and severity of displacement was recorded for each tooth affected by migration. The types of pathologic migration recorded were diastema, extrusion, rotation, facial flaring, and drifting into edentulous spaces. Pathologic migration prevalence was 30.03% +/- 2.5 (103/343 subjects). The mean AL of migrated teeth (4.79 +/- 0.28 mm) was significantly greater (P < 0.0001) than control teeth (3.21 +/- 0.18 mm). The numeric values (called PTV) of migrated teeth (17.6 +/- 1.5) were significantly greater (P < 0.0001) than control teeth (9.4 +/- 1.1). It was difficult to identify a primary form of displacement, as most patients demonstrated a combination of movements. The percentage of the 44 patients who presented with a specific type of movement was: facial flaring (90.9 +/- 4.4%), diastema (88.6 +/- 4.8%), rotation (72.7 +/- 6.8%), extrusion (68.2 +/- 7.1%), and tipping (13.6 +/- 5.2%). The results of this study confirms clinical impressions that periodontal disease destruction of the attachment apparatus plays a major role in the etiology of pathologic migration. PMID- 9358364 TI - Putative periodontal pathogens in subgingival plaque of young adults with and without early-onset periodontitis. AB - This study assessed the associations between putative periodontal pathogens and early-onset periodontitis (EOP) in a population of 248 subjects, 13 to 19 years of age at baseline, derived from a representative sample of U.S. young adults. The subjects were selected based on the presence or absence of attachment loss at baseline. The attachment level was assessed clinically at baseline and at a 6 year follow-up examination, and the presence of 7 bacterial species was assessed at follow-up using DNA probes. The individuals were classified into generalized, localized, incidental EOP, and no-periodontitis groups based on the extent and severity of attachment loss; and classified as having rapid, moderate, slow, and no progression based on the rate of periodontal progression during the 6 preceding years. In the EOP groups there were significantly higher percentages of individuals with detectable levels of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, and Treponema denticola. In addition, the EOP group had significantly higher levels of these 5 microorganisms compared to the no-periodontitis group. There were also higher percentages of individuals with these species and higher levels of bacteria in the group showing disease progression than the group without progression. In a descending order of importance, P. gingivalis, T. denticola, and P. intermedia were the microorganisms significantly associated with the generalized and/or rapidly progressing disease. F. nucleatum and C. rectus were also associated with EOP, but to a lesser degree. In the present population Actinobacillus actinomycetemcomitans was not significantly associated with EOP, though it was recovered more often from subjects with localized EOP. Eikenella corrodens was present equally in subjects with and without disease. The results show that several bacterial species are associated with EOP, and that P. gingivalis and T. denticola are of particular importance and may play a significant role in the more severe and progressive forms of EOP. PMID- 9358365 TI - Guided tissue regeneration using a bioabsorbable membrane in the treatment of human buccal recession. A re-entry study. AB - A surgical technique involving a bioabsorbable membrane was used to treat localized buccal recession defects of between 2 and 5 mm on 17 sites in 13 patients. A conventional flap for coronal repositioning was raised consisting of a trapezoid flap with de-epithelialization of papillae, and the root surface planed back to reduce its convexity. The polylactate membrane with internal spacer bars was placed over the root surface with its coronal edge as near to the cemento-enamel junction (CEJ) as possible. This was held in place by a bioabsorbable retaining suture around the root, and the flap was coronally repositioned to cover the membrane. The patients were prescribed tetracycline for 2 weeks postsurgically, the sutures removed after 4 weeks, and cleaning procedures in the area reinstated after 6 weeks. Measurements were made using a custom made stent, and subjected to a Wilcoxon signed ranked analysis. These were clinical recession, clinical attachment level, probing depth, width of attached gingiva and the level of the buccal alveolar crest. The healing was monitored for a minimum of 1 year, and a surgical re-entry procedure was performed on 11 sites. The new buccal alveolar crest position was measured. This procedure resulted in a clinical and statistically significant reduction in clinical recession of 2.4 +/- 0.2 mm, a gain in attachment level of 2.7 +/- 0.2 mm, a gain in root coverage of 76 +/- 6% (P < 0.002), and a regeneration of buccal bone in all cases, with an average of 2.0 +/- 0.1 mm (P < 0.0023). This study shows that the use of a bioabsorbable membrane will predictably and significantly increase root coverage and regenerate buccal bone when used to treat localized buccal recession defects. PMID- 9358366 TI - Gingival microcirculation response to tooth brushing measured by laser Doppler flowmetry. AB - This study quantified changes in blood flow following tooth brushing, using laser Doppler flowmetry (LDF). Twenty subjects had polysiloxane stents fabricated with openings to permit placement of the LDF probe on the mesial papillae of 6 teeth. Probing depth, plaque index, and gingival index were recorded and subjects instructed in brushing. LDF initially recorded a 30 second baseline blood flow. The stent was removed and subjects brushed the site for either 3 or 10 seconds. The stent was repositioned and recordings again taken, followed by a control reading. The process was repeated 4 weeks later. Correlations between baseline and control readings were 0.585 (P < 0.001) at the first visit, and 0.654 (P < 0.001) at the return visit. The mean control blood flow was 156.4 perfusion units. The 3 and 10 second brushing increased the mean value 22.6 units and 21.2 units respectively (both P < 0.001). Tooth brushing for both 3 and 10 seconds significantly increased gingival blood flow in the papillary gingiva of healthy individuals. PMID- 9358367 TI - Integrated connective tissue in bioabsorbable barrier material and periodontal regeneration. AB - The objective of this study was to evaluate the relationship between integrated connective tissue (ICT), that is, the presence of connective tissue into the membrane structure, and the clinical outcome of membrane-supported periodontal surgery. Twenty-six systemically healthy subjects affected by chronic adult periodontitis were enrolled in the study. One tooth site per patient, associated with an angular bony defect and an attachment loss of > 7 mm, was selected to be treated by means of a guided tissue regeneration procedure using a bioabsorbable membrane. Barrier material was surgically removed after 4 weeks for SEM analysis. For each treated site, the difference in clinical attachment loss, probing depth, and gingival recession between the baseline examination and follow-up 6 months after the second surgery was calculated. Gain of attachment was statistically (P < 0.001) greater in sites with no membrane exposure when compared to sites with exposed barrier material (5.5 +/- 1.0 vs. 4.0 +/- 0.6), while further gingival recession was greater (3.0 +/- 0.9 vs. 2.1 +/- 0.5) in sites with clinically exposed membranes. The results of SEM analysis revealed that when connective tissue structures were observed on membrane surfaces, no bacteria could be detected; conversely, areas heavily colonized by bacteria did not show the presence of connective tissue. Regression analysis indicated that integrated connective tissue on the external layer of the barrier material was positively correlated with the amount of attachment gain and negatively with the amount of gingival recession. Bacterial colonization of the membrane was negatively correlated with attachment gain and positively with gingival recession. It was concluded that connective tissue integration is an important biological phenomenon in preventing membrane exposure and bacterial plaque colonization and thus in enhancing the clinical outcome following guided tissue regeneration surgery. PMID- 9358368 TI - Distribution of metallothionein in cigarette smokers and non-smokers in advanced periodontitis patients. AB - This study evaluated the effect of smoking on periodontal tissue in periodontal patients. Gingival biopsies were taken from the flap during periodontal surgery of 33 male patients with advanced periodontal disease (22 cigarette smokers; 11 non-smokers). Frozen sections were made immediately and used for hematoxylin eosin and Giemsa staining, and for detection of metallothionein (MT), a free radical scavenger. The expression and distribution of MT in these sections was studied using monoclonal antibody and immunohistochemical staining. The smokers had a large number of infiltrate cells in the gingival epithelium compared with non-smokers. Moreover, samples from the smokers showed high levels of MT in the prickle cell layer of the epithelium. In the non-smokers, basal and prickle cell layers of the gingival epithelium also showed detectable MT. Smokers, however, had a higher metallothionein-positive cell ratio in the prickle cell layer as compared to non-smokers. This increased level of MT in smokers may reflect an attempt to defend against free radicals in the gingiva of smokers. These results suggest that inflammation is much greater in periodontal tissues of cigarette smoking periodontal patients than in non-smoking periodontal patients, and that the defense against free radicals is heightened in smokers' gingiva. PMID- 9358369 TI - Differential attachment of oral treponemes to monolayers of epithelial cells. AB - This in vitro study describes the attachment properties of several oral treponemes to monolayers of epithelial cells and the effect of epithelial cell confluence on treponeme attachment. Four serotypes of Treponema denticola, Treponema scoliodontum, three subspecies of Treponema socranskii, and Treponema vincentii were tested with monolayers of epithelial cells of human and canine origin. Attachment of oral treponemes were compared to attachment by T. pallidum subsp. pallidum, and by the non-pathogen Treponema phagedenis. Results indicated that different serotypes of T. denticola had similar abilities to attach to epithelial cells. However, subspecies of T. socranskii differed in their ability to attach to epithelial cells. The proportion of epithelial cells susceptible to attachment by oral spirochetes was strongly related to the confluence level of the monolayer. In contrast, T. pallidum attached equally well to both epithelial cell lines at all confluence levels. T. phagedenis attached to < 1% of all epithelial cells. In general, attachment of oral treponemes to canine cells was lower than to human cells, suggesting species-specificity for adherence. Attachment of oral treponemes to epithelial cells may promote colonization of the periodontal pocket, as well as retention of treponeme colonies within plaque. The preference of oral treponemes to attach to cells of low confluence fields may translate in vivo to an increased ability to attach to cells which are actively dividing. Such cells are found in areas of repair, a common status within inflamed periodontal pockets. Furthermore, attachment of oral treponemes to epithelial cell barriers may promote or potentiate cytopathic processes. PMID- 9358370 TI - Traumatic bone cyst resembling apical periodontitis. AB - Among the pseudocysts of the jaws, the traumatic bone cyst is known as an asymptomatic lesion often noted unintentionally during routine radiographic examinations. The lesion neither devitalizes the teeth within its borders, nor does it cause resorption of their roots. The well-demarcated traumatic bone cyst often projects into the intraradicular septa and hence has been described as having scalloped borders. The following presentation is of a traumatic bone cyst that resembled periodontal pathology in its appearance. PMID- 9358372 TI - Re: Non-surgical treatment of patients with periodontitis. PMID- 9358371 TI - Re: Epidemiology of periodontal diseases (position paper)(1996;67:935-945) PMID- 9358373 TI - Adolescents are the orphans of immunization practices. PMID- 9358374 TI - The importance of adolescent immunization recommendations. PMID- 9358375 TI - The commitment to keeping adolescents healthy. PMID- 9358376 TI - Immunization services for adolescents within comprehensive school health programs. AB - Every school day, more than 95% of the nation's five- to 17-year-olds attend one of almost 110,000 elementary and secondary schools where they receive instruction in how to avoid major health-threatening behaviors. School is also the place where children and adolescents can be given preventive health care services, including administration of vaccines and monitoring of immunization levels. This article addresses how expanding school health services could improve immunization levels of school-aged youth. PMID- 9358377 TI - A multi-disciplinary curriculum for 11- to 13-year-olds: immunization, plus! AB - A sixth grade curriculum entitled "Immunization, Plus!" Was developed to promote adolescent immunization. This targeted immunization curriculum utilized contemporary learning theory and innovative teaching approaches and styles to maximize acceptability among educators. Because instructional time in school was limited, a thematic curriculum was created to embed immunization and communicable disease content within mathematics, science/health, and language arts units. The curriculum, which reflected the theory of multiple intelligences among students, offered an array of different learning formats, including linguistic, logical mathematical, spatial, and bodily-kinesthetic. The curriculum was made available free of charge to school districts in California, and its evaluation was planned to track distribution, utilization, and changes in students' knowledge, attitude, and behavior. PMID- 9358378 TI - Consent for adolescent vaccination: issues and current practices. AB - To identify and describe implementation of state-level informed consent requirements for adolescent immunizations, current state regulations on informed consent and immunization services for children and adolescents were identified through the LEXIS-NEXIS legal data base. Regulations were coded for informed consent characteristics, consent exemptions, and current immunization requirements. State immunization program directors, project managers, and state hepatitis coordinators were surveyed to catalogue how regulations were implemented and document new policies or regulations under consideration. Parental consent for immunizations is standard practice in 43 states. Most states (n = 34) require separate consent for each injection when more than one injection is required to complete a vaccination, but only for a limited number of medical procedures. Nine states allow adolescents to self-consent for hepatitis B vaccination in sexually transmitted disease clinics and family planning clinics as part of the exemption for minors' receipt of sexual health services. Most states require consent for vaccination services provided to adolescents. Parental consent requirements are a potential barrier to vaccinating adolescents in some settings. PMID- 9358379 TI - Incentives and motivators in school-based hepatitis B vaccination programs. AB - During a school-based vaccination program, incentives and education were offered to help motivate students to participate. Each student at all schools in the program received scholastic credit for returning a signed form, material rewards for receiving each vaccine dose, and free attendance at a social event after completing the vaccine series. In two of four schools, classes received a reward if every student in the classroom returned a signed form within five days: in these schools, 91% and 98% of students returned signed forms within five days, compared to 82% and 85%, respectively, in the two schools without this peer incentive. Approximately half the students receiving the peer incentive reported that it played a motivating role, whereas 60% cited wanting to be protected. Few students named individual rewards as motivators. Although peer incentives appeared effective in encouraging some students to return parent consent or refusal forms, the desire to be protected may have been a stronger motivator. PMID- 9358380 TI - Immunizations from ground zero: lessons learned in urban middle schools. AB - The Centers for Disease Control and Prevention funded a three-year demonstration project in San Francisco to assess the feasibility of a large-scale school-based vaccination effort. The project overcame a number of barriers, including lack of pre-existing health services, diversity of home languages, and an every-50-minute bell schedule. The project targeted seventh graders and all special education students for hepatitis B vaccine (HBVac). Of 4,928 students targeted, 3,509 (71%) consented to vaccination and received the first dose. Of these 3,509 students, 3,256 (93%) completed the three-dose series at school. Key lessons learned include emphasizing a collaborative process in the planning stage, offering an educational component for students, providing an incentive to get timely parental consent, planning distribution and collection of parent materials, and planning vaccination clinics to minimize interrupting the school day. The project clearly demonstrated that, with sufficient attention to political and logistical dimensions, school-based vaccination programs are possible in large urban schools. PMID- 9358381 TI - A state-based immunization campaign: the New Mexico experience. AB - Hepatitis B prophylaxis in the form of immunization has become an increasingly effective protective measure across the United States. Various cohorts have been targeted and methodologies used by public health officials to attain optimal coverage of the population. New Mexico is in its third year of school-based hepatitis B immunization programs. New Mexico's approach to this issue is characterized by significant rates of participation (66% in middle schools and 56% in high schools), high vaccination series completion (88% in middle schools and 89% in high schools), and minimal financial investment (cost of vaccine). The "Roll Up Your Sleeves" campaign was piloted, modified, and adapted to New Mexico's multi-lingual and multi-cultural environment, resulting in these successes. PMID- 9358382 TI - Asian-American adolescent immigrants: the New York City schools experience. AB - This article describes development and implementation of a school-based vaccination program that targeted Asian-American adolescents. The program was implemented by the Chinatown Health Clinic in New York City in two high schools and a junior high school in Lower Manhattan. The article examines strategies effective in vaccinating this varied group of adolescents. Rates of completion for the three-dose hepatitis B vaccine are compared between schools. Optional serology testing for hepatitis B infection was conducted as part of the school program and rates of infection in this population are presented. The article discusses the importance and effectiveness of school-based programs in providing essential health services to this group of adolescents. PMID- 9358383 TI - From university to community: the Baton Rouge experience. AB - This paper describes a successful hepatitis B vaccination program which expanded from one school vaccinating 475 students to 68 schools vaccinating 3,400 students. Issues associated with success include acquiring resources, organizing program logistics, developing an advisor board, determining eligibility for free vaccine, obtaining vaccine for those not eligible for publicly supplied vaccine, methods of consenting, and managing data. The process of obtaining support can increase public awareness and assist further program expansion. The program demonstrates that the time and energy expended in start-up activities for the first two or three years will evolve into a program with a life of its own, requiring only a moderate amount of attention while generating strong positive community support. PMID- 9358384 TI - Health education lessons learned: the H.A.P.I. Kids Program. AB - Challenges exist for effective health communication and health education within diverse populations of the United States. This article addresses the development process for educational materials and lessons learned from the Healthy Asian and Pacific Islander (H.A.P.I.) Kids Program, a vaccination demonstration project funded by the Centers for Disease Control and Prevention to promote catch-up hepatitis B vaccination for older American Asian and Pacific Islander children. Simplicity and a common message were incorporated in multiple strategies to disseminate information to a diverse population. Community representatives from the Cambodian, Hmong, Filipino, Lao, and Vietnamese communities were instrumental in the material development process, which included needs assessment, design, and translation. By making the target community part of the development process, important health messages can be disseminated effectively, carrying great impact to an otherwise hard-to-reach community. PMID- 9358385 TI - Managed care organizations and public health: exploring collaboration on adolescent immunizations. AB - Managed care organizations (MCOs) joined local and state public health agencies in a pilot effort to improve hepatitis B immunization rates of adolescents in an urban and a suburban/rural school district. The pilot also explored issues inherent in public and private collaboration on population health improvement. Local public health agencies provided links to schools in their communities, took the lead in implementing school-based immunization programs, and provided health education materials. MCOs contributed financial support necessary for the project. The final cost per fully vaccinated student, not taking into account the work group's planning and coordination time, was little more than the catalog price of the vaccine alone. Managed care organizations face challenges that complicate their participation and funding of school-based vaccinations: 1) Limited data on health plans of participating students complicate allocation of costs to each MCO; 2) Double-paying occurs for MCOs paying clinics a monthly, per member rate that already includes adolescent immunizations; 3) When schools provide adolescent immunizations, MCOs lose the "hook" that draws adolescents to clinics for comprehensive health services. When self-consenting is permitted, schools can achieve a high consent and completion rates for multi-dose adolescent immunizations such as hepatitis B. At the same time, MCOs have the responsibility to provide members with comprehensive care and should continue to examine both internal modifications and external partnerships as opportunities to improve their services to adolescents. PMID- 9358386 TI - Beyond band aids: school health nurses as program developers and coordinators. PMID- 9358387 TI - Parent involvement in health concerns for youth: the issue of adolescent immunization. AB - Parents play a primary role in the health and health education of their children. In particular, parent involvement in planning and promoting adolescent immunization campaigns is critical to successful efforts. Parents serve as their children's primary educators on health issues, but where can they get accurate health information? To help guide local PTA units in their programmatic efforts, the National PTA maintains positions and policy statements on multiple health issues: alcohol and other drug abuse; emergency preparedness; environmental issues; family life education; firearm safety; HIV prevention; health screenings; immunization (measles, mumps, rubella, and hepatitis B); lead poisoning; nutrition; protective helmet use; sexual assault prevention; TB testing; tobacco use and access; violence prevention; and youth suicide prevention. Likewise, the school-home partnership is key to promoting the health of adolescents. Comprehensive school health programs and integrated services are necessary to support parent and community efforts to promote adolescent health issues, including immunization programs. Techniques for effective parent involvement, based on the National Standards for Family/Parent Involvement issued by the National PTA January 1997, are discussed. PMID- 9358388 TI - From formal to friendly: creating materials that work with adolescents. AB - The Immunization Action Coalition, publisher of over 100 health educational pieces on immunization, offers a way to take a formal recommendation to vaccinate all 11- and 12-year-olds against hepatitis B by creating a friendly educational piece that can make adolescents and their parents sit up, take notice, roll up their sleeves, and run to doctors to demand their hepatitis B shots. The brochure created follows dictates of passion, words, graphics, feedback, and no copyright. The coalition's semi-annual newsletter, Needle Tips & the Hepatitis B Coalition News, encourages adaptation or use of the brochure, which is approved by the Centers for Disease Control and Prevention. PMID- 9358389 TI - Promoting hepatitis B vaccinations for adolescents: the use of advocacy organizations. PMID- 9358390 TI - Immunization of adolescents: recommendations of the Advisory Committee on Immunization Practices, the American Academy of Pediatrics, the American Academy of Family Physicians, and the American Medical Association. AB - This report concerning the immunization of adolescents (ie, persons 11-21 years of age, as defined by the American Medical Association [AMA] and the American Academy of Pediatrics [AAP]) is a supplement to previous publications (ie, MMWR 1994;43[No. RR-1] 1-38; the AAP 1994 Red Book: Report of the Committee on Infectious Diseases; Summary of Policy Recommendations for Periodic Health Examination, August 1996 from the American Academy of Family Physicians [AAFP]; and AMA Guidelines for Adolescent Preventive Services [GAPS]: Recommendations and Rationale). This report presents a new strategy to improve the delivery of vaccination services to adolescents and to integrate recommendations for vaccination with other preventive services provided to adolescents. This new strategy emphasizes vaccination of adolescents 11-12 years of age by establishing a routine visit to their health-care providers. Specifically, the purposes of this visit are to a) vaccinate adolescents who have not been previously vaccinated with varicella virus vaccine, hepatitis B vaccine, or the second dose of the measles, mumps, and rubella (MMR) vaccine; b) provide a booster dose of tetanus and diphtheria toxoids; c) administer other vaccines that may be recommended for certain adolescents; and d) provide other recommended preventive services. The recommendations for vaccination of adolescents are based on new or current information for each vaccine. The most recent recommendations from ACIP, AAP, AAFP, and AMA concerning specific vaccines and delivery of preventive services should be consulted for details. PMID- 9358391 TI - Adolescent immunization: focus on implementation. AB - On March 11-12, 1996, a workshop on how to implement new adolescent immunization (AI) recommendations was held in Atlanta, Ga. Sponsored by the Centers for Disease Control and Prevention, it was a collaborative effort of the National Immunization Program, the Division of Adolescent and School Health/National Center for Chronic Disease Prevention and Health Promotion, and the Hepatitis Branch/National Center for Infectious Diseases. The workshop brought together organizations and individuals interested in adolescent health and immunizations so they could address how new AI recommendations can be implemented most effectively. This article offers an overview of their discussions and suggestions, including issues of cooperation, education, legislation, and AI program development among health provider organizations, health department, schools, community groups and various other agencies relating to adolescent health services. PMID- 9358392 TI - Topographic distribution of sleep spindles in young healthy subjects. AB - The application of an automatic sleep spindle detection procedure allowed the documentation of the topographic distribution of spindle characteristics, such as number, amplitude, frequency and duration, as a function of sleep depth and of recording time. Multichannel all-night EEG recordings were performed in 10 normal healthy subjects aged 20-35 years. Although the interindividual variability in the number of sleep spindles was very high (2.7 +/- 2.1 spindles per minute stage 2 sleep), all but two subjects showed maximal spindle activity in centro-parietal midline leads. Moreover, this topography was seen in all sleep stages and changed only slightly--to a more central distribution--towards the end of the night. On the other hand, slow (11.5-14 Hz) and fast (14-16 Hz) spindles showed a completely different topography, with slow spindles distributed anteriorly and fast spindles centro-parietally. The number of sleep spindles per min was significant depending on sleep stages, with the expected highest occurrence in stage 2, and on recording time, with a decrease in spindle density from the beginning towards the end of the night. However, spindle amplitude, frequency and individual duration was not influenced by sleep depth or time of the night. PMID- 9358393 TI - Endogenous and exogenous components in the circadian variation of core body temperature in humans. AB - Core body temperature is predominantly modulated by endogenous and exogenous components. In the present study we tested whether these two components can be reliably assessed in a protocol which lasts for only 120 h. In this so-called forced desynchrony protocol, 12 healthy male subjects (age 23.7 +/- 1.4 y) were subjected one by one to an artificial light/dark cycle of 20 h (10 lux vs. darkness). Core body temperature was measured continuously. The temperature data were analysed by an iterative method based on the assumption that the endogenous and exogenous components contribute to body temperature in an additive way. The results show that the average temperature curve is an almost perfect addition of the two components. The endogenous component differs from a sinusoid, and the relative contributions of the endogenous and exogenous components to the raw temperature curves differ substantially between the subjects. The average amount of unexplained variance in the individual data was 17%. Averaging of the body temperature curves over subjects reduced the unexplained variance to only 2%. This reduction in unexplained variance upon averaging over subjects must be due to the fact that most of the variance is either differently dependent on circadian phase for the various subjects or not dependent on circadian phase at all. The circadian pacemaker component revealed an average value of tau of 24.30 +/- 0.36 h, which is consistent with recent findings in the literature. We conclude that a short forced desynchrony protocol is sufficient for the distinction between the masking and pacemaker components of core body temperature. The same protocol can be used to study the influence of these components on all kinds of other physiological and psychological signals. PMID- 9358394 TI - Effect of intracerebroventricular administration of alpha-helical CRH (9-41) on the sleep/waking cycle in rats under normal conditions or after subjection to an acute stressful stimulus. AB - It has been shown in a previous study that specific lesioning of the noradrenergic system of the locus coeruleus abolished the sleep increase induced by immobilization stress. Given the fact that brain corticotropin-releasing hormone (CRH) acts as a neurotransmitter in the locus coeruleus under stress conditions, the present study was designed to investigate the involvement of CRH in the sleep increase seen after immobilization stress and on the spontaneous wake/sleep cycle. One hundred micrograms of the specific CRH receptor antagonist alpha-helical CRH (9-41), or vehicle alone was injected into the right lateral ventricle 30 min either before subjecting the animals to immobilization stress or before the spontaneous sleep-waking recordings onset. A single intracerebroventricular (i.c.v.) injection of alpha-helical CRH (9-41) had no effect on spontaneous paradoxical sleep but abolished the stress-induced increase, while wakefulness and slow-wave sleep were unchanged under both normal and stressful conditions. We therefore report that the involvement of endogenous CRH in the paradoxical sleep mechanism is dependent on the environmental conditions and suggest that, while the paradoxical sleep increase induced by immobilization stress may be mediated by endogenous corticotropin-releasing hormone, other mechanisms, either CRH-independent or situated at a distance from antagonist activity, may be involved in spontaneous paradoxical sleep. These results show, for the first time, that endogenous CRH may be involved in sleep waking mechanisms only under stressful conditions and, in particular, as a fundamental component of the paradoxical sleep increase. PMID- 9358395 TI - Sleep deprivation increases somatostatin and growth hormone-releasing hormone messenger RNA in the rat hypothalamus. AB - We studied the effect of sleep deprivation (SD) on the amount of somatostatin (SRIF) and growth hormone-releasing hormone (GHRH) mRNA in rat hypothalamic nuclei. According to earlier studies SRIF possibly facilitates REM sleep and GHRH slow-wave sleep. Adult male rats were sleep deprived by the gentle handling method either for 6 h during the first half of the light phase or for 12 h during the dark phase. Undisturbed rats sacrificed at the same time as the SD rats served as controls. After oligonucleotide in situ hybridization the amount of SRIF and GHRH mRNA was measured in brain sections by image analysis and cell count. SD increased the amount of SRIF mRNA in the arcuate nucleus (ARC). In the periventricular nucleus (PE) there was no effect. The amount of GHRH mRNA increased in the paraventricular nucleus (PA) in the 6 h SD group but no effect was detected in ARC. In the periventromedial hypothalamic area (pVMH) the amount of GHRH mRNA was higher in the control rats sacrificed in the morning (09.00 hours) than in the afternoon (15.00 hours), and SD had no effect. We conclude that SRIF cells in ARC and GHRH cells in PA are modulated by sleep loss, which is in accordance with the possible sleep regulatory function of these neuropeptides. PMID- 9358396 TI - Psychophysiological insomnia: the behavioural model and a neurocognitive perspective. AB - A number of paradoxes are apparent in the assessment and treatment of psychophysiological insomnia and sleep state misperception. Three of these paradoxes exist as discrepancies between polysomnographic (PSG) measures and the subjective impressions regarding sleep quality and quantity. The remaining incongruity exists largely within the objective domain. In the case of subjective objective discrepancies, patients with insomnia: (1) frequently identify themselves as having been awake when awakened from PSG defined sleep; (2) tend to overestimate sleep latency and underestimate total sleep time as compared with PSG measures; (3) appear to derive more benefit from pharmacotherapy that can be explained by objective gains. The remaining paradox pertains to the observation that hypnotic medications, by and large, do not normalize sleep architecture or produce a more 'sleep-like' EEG. In this paper, we review possible explanations for these various paradoxes, introduce a new perspective and suggest possible research avenues. The model introduced is based on the observation that beta and/or gamma activity (which have been found to be associated with cognitive processes) is enhanced in insomnia at or around sleep onset. We propose that this kind of high frequency EEG activity may interfere with the normal establishment of sleep onset-related mesograde amnesia. As a result, the patient with insomnia maintains a level of information and/or memory processing that blurs the phenomenological distinction between sleep and wakefulness and influences retrospective judgments about sleep initiation and duration. PMID- 9358397 TI - Perceived tiredness among adolescents and its association with sleep habits and use of psychoactive substances. AB - This study investigated the variation in perceived tiredness among 11, 13 and 15 year-old Finnish adolescents (n = 4187). Additionally interrelationships between sleep habits, use of psychoactive substances (alcohol, tobacco and coffee) and perceived tiredness among 15-year-olds were examined. This study is part of an international, WHO-coordinated survey of school children's health and lifestyle (the HBSC Study). In Finland, research data represented the whole country. The data were collected in March-May 1994. Pupils responded anonymously to a standardized questionnaire during a class period. Subjective tiredness was very common and increased with age among adolescents. Perceived tiredness on at least four school mornings a week increased from 24 to 35% among boys and from 16 to 34% among girls. Feeling tired more often than once a week increased from 20 to 37% in girls and from 24 to 50% in boys. Structural equation models showed that interrelationships between three factors--sleep habits, use of psychoactive substances and perceived tiredness--were statistically significant. For these three factors the two remaining factors explained 24% of the variance of perceived tiredness among boys and 20% among girls, and the two remaining factors explained 42% (16%) of the variation in sleep habits. For the variance of the use of psychoactive substances sleep habits and perceived tiredness explained 26% (12%). Subjective tiredness is strongly age related; this together with the use of psychoactive substances and sleep habits regulate adolescents' daily life and well-being. PMID- 9358398 TI - Comparison of three measures of quality of life outcome in the evaluation of continuous positive airways pressure therapy for sleep apnoea. AB - Treatment of obstructive sleep apnoea (OSA) with nasal continuous positive airway pressure (NCPAP) has become a standard treatment since its introduction in 1981. Following such treatment the apnoeas disappear, sleep quality improves as apparently do daytime symptoms of sleepiness. Sleepiness is an unusual symptom and its impact on conventional indices of quality of life has rarely been measured. To allow comparison of NCPAP therapy with treatments for other conditions, measurements of quality of life before and after treatment using standard techniques are required. It is not clear which of the standard measures is most suited to measuring the health gain from nasal NCPAP, and indeed whether the disability of excessive sleepiness is included in all such measures. This study looks at three well recognized quality of life measures in OSA, before and after NCPAP therapy; the Short Form 36 (SF-36), Functional Limitations Profile (FLP), and the EuroQol (EQ-5D). The results were compared with data from normal populations. One hundred and eight patients with OSA undergoing a therapeutic assessment of NCPAP completed the three quality of life questionnaires before and 5 weeks after commencing treatment. Over 90 subjects completed all sections of the three measures on both occasions. The SF-36 revealed substantial adverse effects on subjective health of OSA and that NCPAP treatment produced dramatic positive effects. For example, the effect sizes (difference in score, divided by SD of baseline score) in the Energy/Vitality dimension was 0.98 and for the overall Mental and Physical Component Scores, 0.76 and 0.57, respectively: an effect size over 0.5 is considered moderate and over 0.8 as large. The FLP data showed similar pre treatment decrements in quality of life and substantial improvements following NCPAP. The changes with treatment in the majority of the dimensions from both the SF-36 and FLP were statistically significant (P < 0.01). In contrast the EQ-5D did not show significant improvements with therapy, presumably because of its failure to measure the aspects of quality of life related to severe sleep fragmentation and daytime sleepiness. In conclusion, this study has clearly shown considerable decrements in quality of life in patients with OSA, similar to other chronic disabling conditions. Furthermore, NCPAP therapy returns patients to a quality of life similar to the normal population. PMID- 9358399 TI - Metamemory in narcolepsy. AB - People with narcolepsy consistently report diminished memory function attributable to the disorder, however, objective evaluations of memory performance in this clinical group remain inconclusive. Previous evaluations of these subjective experiences have been primarily anecdotal with subjects required to provide global assessments of their memory function. The present study aimed to evaluate subjective assessments of memory dysfunction more extensively comparing responses by narcoleptics, subjects experiencing excessive daytime sleepiness, and controls, on the Metamemory in Adulthood (MIA) questionnaire. The results of the study indicate that subjects with narcolepsy have lower self efficacy for memory performance than either of the comparison groups, despite there being no significant difference between groups in relation to knowledge based aspects of memory functioning. This lowered self efficacy in narcolepsy is expressed through increased anxiety about memory function, decreased evaluations of memory capacity and increased perceptions of memory decline in relation to the comparison groups. It is argued that the negative cognitive self evaluations of narcoleptics potentially arise as a consequence of global psychosocial adjustment difficulties. PMID- 9358400 TI - Slow-wave sleep: do young adult men and women age differently? AB - The differential effects of ageing on polysomnographic and EEG spectral characteristics of sleep were explored in men and women between the ages of 20 and 40. Men and women in their twenties were found to have similar percentages of slow-wave sleep (SWS) (% Stage 3 and 4) and mean EEG slow wave activity (quantified by spectral analysis). Significant reductions in the percentage of SWS and mean slow wave activity over the night occurred in men during their thirties but not in the women. This suggests that gender difference in SWS may emerge between age 30 and 40 in young adults. Men in this sample were also found to have significant increases in Stage 2 sleep, and decreases in REM sleep time, REM activity, REM density and REM intensity. No significant effects of age were found for women in any visually scored sleep variables. Both men and women had age related reductions in spectral power in the spindle frequencies. Taken together, these findings suggest that the sleep of men and women over age 20-40 may age differently. PMID- 9358401 TI - What is your diagnosis? Ethylene glycol intoxication. PMID- 9358402 TI - Pharmacokinetics of a porcine insulin zinc suspension in diabetic dogs. AB - Ten dogs with naturally occurring diabetes mellitus were injected with a highly purified porcine insulin zinc suspension at a dose according to their expected requirement. Plasma insulin and glucose concentrations were measured at two hourly intervals over 24 hours following injection. There were either one or two peaks in plasma insulin concentration: one at about four hours (mean 4.3 +/- 1.3 [SD]) and another at about 11 hours (mean 11 +/- 1.85) after the injection. The second insulin peak was seen in only eight dogs. Persistence of elevated plasma insulin concentrations ranged from 14 to 24 hours (mean 17.4 +/- 3.65). These results compare favourably with those published for other intermediate-acting insulin preparations used to treat canine diabetes mellitus and suggest that this preparation has useful properties for the successful management of many canine diabetics. PMID- 9358403 TI - Ultrasonographic imaging of the tongue and larynx in normal dogs. AB - Ultrasonographic imaging of the tongue and larynx was performed in 10 dogs with no previous history of upper airway disease. The ultrasonographic findings were compared with the normal canine anatomy of this area and with the results described in the human literature. This study shows that the anatomical features of the canine larynx are adequately detectable using ultrasonography. This finding is in accordance with the findings described in the human literature. It is concluded that ultrasonography may offer a means of investigating canine laryngeal abnormalities. PMID- 9358404 TI - Avulsion of the tibial tuberosity in a litter of greyhound puppies. AB - Avulsion of the tibial tuberosity was diagnosed in six of seven greyhound littermates aged five and a half months. The puppies showed hindlimb lameness of varying severity. Radiological assessment of affected stifle joints revealed partial or complete avulsion of the tibial tuberosities. In four puppies the lesions were bilateral. Euthanasia of the two most severely affected puppies was performed; the changes observed on histopathological examination of their cranioproximal tibiae suggested that the underlying lesion was that of osteochondrosis. A hereditary predisposition in greyhounds to osteochondrosis of the physis between the apophysis and the cranioproximal tibial diaphysis is postulated. PMID- 9358406 TI - Arteriovenous fistula involving the prepuce of a dog. AB - A nine-year-old male Ibizan hound had a network of large tortuous pulsating blood vessels on the prepuce that enlarged gradually over a five month period. A diagnosis of arteriovenous fistula was based on clinical signs, angiography and Doppler ultrasonography. Ligation of the major vascular supply to the fistula resulted in only temporary improvement. Definitive treatment was by wide excision, with penile amputation and scrotal urethrostomy. PMID- 9358407 TI - Chronic caecal faecolithiasis in a dog. AB - A 14-year-old, spayed female Jack Russell terrier with a six month history of weight loss, lethargy, intermittent vomition and diarrhoea was diagnosed as having a chronic impaction of the caecum with mineralised faecal material. Diagnosis was based on the clinical findings and both survey and positive contrast radiographic studies. The diagnosis of caecal impaction was confirmed at surgery and a typhlectomy was performed with the aid of a linear stapler. Histopathology of the caecum confirmed the impaction to have resulted from faecolithiasis. The dog made a full recovery from the procedure, showing no recurrence of the clinical signs until euthanasia three months postoperatively for probable heart failure associated with mitral regurgitation. PMID- 9358405 TI - Resolution of exfoliative dermatitis and Malassezia pachydermatis overgrowth in a cat after surgical thymoma resection. AB - A four-year-old, male neutered domestic shorthaired cat was presented with a two week history of nasal and ocular discharge, generalised exfoliative dermatitis, intense pruritus, polydipsia, polyphagia, weight loss, intermittent hindlimb ataxia and lethargy. Cutaneous populations of Malassezia pachydermatis yeast organisms were found to be elevated. The generalised nature of the disease prompted survey radiography which revealed the presence of a cranial mediastinal mass which was subsequently resected and found to be a thymoma. Within six months of surgery, systemic and cutaneous signs had resolved and yeast counts had returned to normal, suggesting a causal relationship between the thymoma and the skin disease. PMID- 9358408 TI - Dorsal radiocarpal ligament sprain causing intermittent carpal lameness in high activity dogs. AB - Five dogs were presented with mild to moderate unilateral thoracic limb lameness attributable to second and third degree sprains of the dorsal radiocarpal ligament. Three were active working dogs and two were racing greyhounds. In four cases the lameness was long-standing and unresponsive to rest; the remaining case was seen as an acute injury. Treatment was by exploratory surgery with postoperative support or by external support alone. All the dogs made a complete recovery with no recurrence of injury. PMID- 9358409 TI - Horner's syndrome following intrathoracic tube placement. AB - Two dogs developed unilateral Horner's syndrome attributable to the placement of a thoracic drainage tube. Signs resolved following removal of the chest drains. PMID- 9358410 TI - Zygomatic salivary cyst with mucocele formation in a cat. AB - An eight-year-old neutered male domestic shorthair cat had a zygomatic salivary cyst with associated mucocele formation. A fluctuating swelling developed ventral to the right eye, causing bulging of the lower eyeild, and a corresponding swelling was present in the caudal vestibule of the oral cavity. Cytological examination of the fluid obtained from aspiration of the swelling was consistent with the appearance of saliva. A tentative diagnosis of zygomatic salivary mucocele was made. Surgical exploration and resection of the swelling demonstrated the presence of a multilobular cystic zygomatic salivary gland. The histopathological appearance of the resected tissue was interpreted as an inflammatory reaction to mucus derived from a multilocular salivary cyst. PMID- 9358411 TI - Echinococcosis. PMID- 9358412 TI - Problems common to older dogs and elderly humans. PMID- 9358413 TI - Nasal aspergillosis: treatment with clotrimazole. PMID- 9358414 TI - The effects of four, commercial ceruminolytic agents on the middle ear. AB - Four, commercially available ceruminolytic agents and physiological saline were screened for ototoxic and inflammatory reactions on the middle ear mucosae of guinea pigs (n = 38) and dogs (n = 24). Each solution was injected transtympanically in anesthetized animals. The effects were assessed by brain stem auditory evoked response (BAER) tests to evaluate hearing function and by histological examination of the middle ear structures. Varying degrees of hearing loss and inflammation were observed in some guinea pigs and dogs treated with solutions A, C, and D, whereas no abnormal finding was associated with solution B or saline. PMID- 9358415 TI - Clinical aspects and surgical treatment of hyperadrenocorticism in the domestic ferret: 94 cases (1994-1996). AB - The signalment, clinical findings, laboratory values, and histopathological results of 96 ferrets with signs (i.e., bilaterally symmetrical alopecia, return to male sexual behavior, or an enlarged vulva) suggestive of hyperadrenocorticism were evaluated retrospectively. Of these 96 ferrets, 94 (98%) were diagnosed with hyperadrenocorticism histologically. Treatment consisted of unilateral adrenalectomy for unilateral tumors (84%) and subtotal bilateral adrenalectomy for bilateral adrenal tumors (16%). The histopathological diagnosis included nodular hyperplasia (56%), adrenocortical carcinoma (26%), and adrenocortical adenoma (16%). Common concurrent diseases included splenomegaly (87%), islet-cell tumor (27%), and cardiomyopathy (10%). PMID- 9358416 TI - X-linked severe combined immunodeficiency in a family of Cardigan Welsh corgis. AB - Two, male, Cardigan Welsh corgi puppies, one of which was diagnosed with X-linked severe combined immunodeficiency (XSCID), are described in this report. The first puppy was euthanized before definitive immunological testing could be performed. When the second puppy was presented and the relationship between the two was discovered, immunological testing was pursued immediately due to this puppy's rapid deterioration. The immunological test results and genetic studies were compared to the XSCID basset hounds and found to be similar. By unveiling the mutation, the pedigree could be analyzed and the carrier females removed from the breeding population. PMID- 9358417 TI - The use of polymeric liquid enteral diets for nutritional support in seriously ill or injured small animals: clinical results in 200 patients. AB - This prospective, multicenter study evaluated the use of four polymeric liquid enteral (PLE) diets manufactured for dogs and cats in 200 ill or injured patients. Polymeric liquid enteral diets were administered by free-choice feeding, syringe, or feeding tube for up to 208 days. Overall results indicated a 4.9% incidence of vomiting in dogs and a 7.9% incidence in cats; an 8.9% incidence of diarrhea in dogs and an 18.4% incidence in cats. Patients fed the PLE diets seven days or longer had an average increase in body weight of 1.4% in dogs, an average decrease in body weight of 3.8% in cats, increases in lymphocyte counts, and mild decreases in serum albumin. PMID- 9358418 TI - Flavobacterium breve meningitis in a dog. AB - An unusual, gram-negative rod was isolated in significant numbers (4+) from the cerebrospinal fluid (CSF) of a dog. This isolate, Flavobacterium breve, has not been identified previously as a pathogen in the dog. The case and the characteristics of the organism are described. PMID- 9358419 TI - Endoscopic retrieval of a large, nasopharyngeal foreign body. AB - A 1.5-year-old golden retriever was presented for stertorous respiration, reverse sneezing, halitosis, and a bilateral mucopurulent nasal discharge. A bone density, nasopharyngeal foreign body was visualized on lateral radiographs of the skull. The foreign body was removed using a videoendoscope and a basket retrieval forcep. The dog's clinical signs resolved following foreign body removal. PMID- 9358421 TI - Gastrinoma: a retrospective study of four cases (1985-1995). AB - Islet-cell tumors of the pancreas, such as gastrinoma, are rare in veterinary medicine. Patients with gastrinoma or Zollinger-Ellison syndrome have elevated serum gastrin levels which ultimately cause gastrointestinal ulcerations. Due to their small size, gastrinomas are a challenge to localize prior to surgery. In veterinary medicine, exploratory surgery with biopsy for histopathology confirms the diagnosis of gastrinoma. This is a retrospective study of four dogs with gastrinoma. PMID- 9358422 TI - Scapular fractures in dogs: epidemiology, classification, and concurrent injuries in 105 cases (1988-1994). AB - A retrospective study of canine scapular fractures diagnosed and treated from 1988 through 1994 at four veterinary teaching hospitals was performed. Dogs (n = 105) with 109 scapular fractures were included. Most scapular fractures occurred in young (i.e., less than four years of age), male, medium- to large-breed (i.e., greater than 10 kg) dogs as the result of vehicular trauma. Concurrent injuries (primarily thoracic trauma) occurred in approximately 70% of cases. In-house follow-up evaluations were considered adequate in only 17% of the cases. A classification system that includes biomechanical principles for categorization is described to avoid discrepancies between various traditional classification systems. PMID- 9358420 TI - Pharmacokinetics and suggested oral dosing regimen of cisapride: a study in healthy cats. AB - The disposition of cisapride in seven healthy cats was determined following administration of either a single oral (2 mg/kg body weight) or intravenous (i.v.) (1 mg/kg body weight) dose. Cats were studied using a random crossover design. After administration of the oral capsule, maximum plasma drug concentration (Cmax) +/- standard deviation (SD) was 73.32 +/- 16.59 ng/ml, and bioavailability +/- SD was 29.0 +/- 22.6%. Following i.v. administration, extrapolated peak cisapride concentration (C0) +/- SD was 421.30 +/- 155.37 ng/ml, and clearance +/- SD was 15 +/- 0.67 ml/kg per minute. Elimination half life (T1/2) was similar for both routes of administration (T1/2(oral) +/- SD was 5.27 +/- 3.16 hr, T1/2(i.v.) +/- SD was 5.19 +/- 3.77 hr). Adverse effects were not observed. Based on these results, a dose of 1 mg/kg body weight per os (PO) every eight hours or 1.5 mg/kg body weight every 12 hours is expected to result in plasma drug concentrations within the therapeutic ranges established for humans. PMID- 9358423 TI - Chronic vaginocervical prolapse with visceral incarceration in a dog. AB - A bitch was presented for a vaginal prolapse of five years' duration. The prolapse was confirmed by physical examination and evaluated by contrast radiography. Herniation of the uterine body, urinary bladder, and distal aspect of the colon was identified within the prolapse. The prolapse was reduced surgically, and an ovariohysterectomy, cystopexy, and colopexy were performed. Compared to other vaginal disorders, vaginal prolapse is an uncommon condition in the bitch. The secondary involvement of abdominal viscera appears to be exceptionally rare. PMID- 9358425 TI - Tetracycline and niacinamide for the treatment of sterile pyogranuloma/granuloma syndrome in a dog. AB - A sterile pyogranuloma/granuloma syndrome in a dog is described. Diagnosis was based on cytological examinations of the skin and lymph nodes and histopathological examinations of the skin and nictitans. Although the condition initially was responsive to large doses of glucocorticoids, it subsequently was treated successfully with tetracycline and niacinamide. The excellent responses of this dog suggest that this drug combination may be a viable therapeutic option for dogs in which glucocorticoids cannot be used. PMID- 9358424 TI - Successful treatment of uterine torsion and fetal retention in a postparturient Great Pyrenees bitch with septic peritonitis and prothrombotic complications. AB - The treatment and favorable outcome of a bitch with uterine torsion and two retained fetuses are described. The condition was corrected surgically by ovariohysterectomy. Complications (i.e., septic shock, peritonitis, and hemostatic abnormalities) were managed with aggressive medical therapy. Torsion of the gravid uterus in dogs is a life-threatening condition which can have a successful outcome if medical complications encountered in the pre- and postoperative periods are treated quickly and effectively. PMID- 9358426 TI - A retrospective study of canine dilated cardiomyopathy (189 cases). AB - The case records of 189 dogs (including 38 breeds) with congestive heart failure caused by dilated cardiomyopathy were studied retrospectively. Airedale terriers, boxers, Doberman pinschers, English cocker spaniels, Newfoundlands, St. Bernards, and standard poodles were over-represented. German shepherd dogs were under represented. A male predominance was observed. Systolic murmurs were detected in 25% of the cases. Atrial fibrillation was the most common arrhythmia. Mild hyperglycemia and mild-to-moderate hypercholesterolemia were found in 38% and 33% of cases, respectively. Histopathological changes consisted of attenuated wavy fibers and interstitial fibrosis. PMID- 9358427 TI - High-resolution nuclear magnetic resonance spectroscopy--applications to fatty acids and triacylglycerols. AB - During the past two decades, nuclear magnetic resonance spectroscopy (NMR) has played an ever-increasing role in the structural determination of fatty acids, fatty acid derivatives and analogues, and in the analysis of the structures of triacylglycerols including the quantitative analysis of lipid mixtures. This article discusses some of the results obtained through the application of the NMR technique to lipid molecules and reviews the literature. To maintain brevity, this article does not cover the underlying theory of NMR spectroscopy as numerous books devoted to modern NMR spectroscopy have been published. PMID- 9358429 TI - Synthesis and nuclear magnetic resonance properties of all geometrical isomers of conjugated linoleic acids. AB - Pure geometric isomers of conjugated linoleic acid were prepared from castor oil as the primary starting material. Methyl octadeca-9Z,11E-dienoate (2) and methyl octadeca-9Z,11Z-dienoate (4) were obtained by zinc reduction of methyl santalbate (1, methyl octadec-11E-en-9-ynoate) and methyl octadec-11Z-en-9-ynoate (3), respectively, as the key intermediates. Methyl octadeca-9E,11E-dienoate (8) and methyl octadeca-9E,11Z-dienoate (9) were prepared by demesylation of the mesyloxy derivative of methyl ricinelaidate (6, methyl 12-hydroxy-octadec-9E-enoate). A study of the nuclear magnetic resonance spectral properties was carried-out, and the shifts of the olefinic carbon atoms of 18:2(9Z,11E) (2) and 18:2(9E,11Z) (9) were readily identified by a combination of incredible natural abundance double quantum transfer experiment, heteronuclear multiple bond correlation, and 1H-13C correlation spectroscopy correlation techniques. Doubts remain in the absolute identification of the individual olefinic carbon atoms of the 18:2(9Z,11Z) (4) and 18:2(9E,11E) (8), except the fact that the shifts of the "inner" (C-10 and C 11) and "outer" (C-9 and C-12) positioned olefinic carbon atoms of the conjugated diene system are distinguishable. PMID- 9358428 TI - Physicochemical characterization of psychosine by 1H nuclear magnetic resonance and electron microscopy. AB - Krabbe's disease is an autosomal recessive disease that affects the lysosomal enzyme galactosylceramidase. The storage of one of its substrates, psychosine (beta-galactosyl-sphingosine), is thought to be responsible for the induction of pathological changes. Psychosine has a free amine group which is necessary for the mediation of its toxic effects. In the present study, the physicochemical properties of psychosine were investigated. Nuclear magnetic resonance (NMR) detected pH titration was used to determine that the amine group had a pKa of 7.18 +/- 0.05. Pulsed-field gradient NMR spectroscopy was used to determine that the diffusion coefficient of 2.8 mM psychosine in D2O at pD 4.46 or 7.04 is 1.16 +/- 0.02 x 10(-10) m2/s or 0.77 +/- 0.02 x 10(-10) m2/s, respectively. Negative staining electron microscopy (EM) studies of acidic and neutral solutions of psychosine also were performed. At pH 4.5, spherical structures were formed, which were relatively stable between 3, 120, and 216 h following preparation; the diameter ranged from approximately 14 nm at the earliest time point to approximately 18 nm at the last time point. The critical micelle concentration (CMC) was 1.26 mM at pH 4.0. At pH 7.1, the structures changed from spherical structures with a diameter of 15-23 nm, at the earliest time point, to a heterogeneous population of structures ranging from spherical structures, with a diameter of only a few nm, to irregularly shaped oblong structures that had one or more dimensions exceeding 100 nm. The NMR and EM data indicate that the deprotonation of the amine group causes psychosine to form aggregates that are unstable, which prevents a determination of the CMC at a neutral pH. These data indicate that molecular interactions of psychosine at the acidic pH of the lysosome, where it is normally digested, are more orderly than those at the pH of the cytoplasm or extracellular space where psychosine goes during disease. PMID- 9358430 TI - Growth inhibitory effects of liposome-associated 1-O-octadecyl-2-O-methyl-sn glycero-3-phosphocholine. AB - The growth inhibitory effects of 1-O-octadecyl-2-O-methyl-sn-glycero-3 phosphocholine (ET-18-OCH3) and various liposome compositions of ET-18-OCH3 were compared in a standardized growth inhibition assay utilizing a diverse tumor cell line panel including cell lines expressing multidrug resistance. ET-18-OCH3 and ELL-12 (4:3:1:2, dioleoylphosphatidylcholine/ cholesterol/dioleoylphosphatidylethanolamine-glutaric acid/ET-18-OCH3), an optimal liposomal ET-18-OCH3 formulation, inhibited growth in the micromolar range in drug-sensitive and -resistant cells. In general, ET-18-OCH3-liposomes were about twofold less growth inhibitory than ET-18-OCH3. However, the known hemolytic effects of ET-18-OCH3 were greatly reduced, up to 20 or more times, by liposome association. The effects of ET-18-OCH3 and ELL-12 were compared in intracellular [Ca2+] modulation and DNA fragmentation assays. ET-18-OCH3 elicited both concentration- and serum-dependent transient and permanent increases in intracellular [Ca2+]. In contrast, ELL-12 did not modulate intracellular [Ca2+]. ET-18-OCH3 and ELL-12 similarly affected DNA fragmentation, which may be indicative of apoptosis. The results suggest that, although the specific growth inhibitory effects of ET-18-OCH3 and ELL-12 are similar, associating ET-18-OCH3 with stable well-characterized liposomes eliminates nonspecific cell membrane associated lytic effects. PMID- 9358431 TI - In vitro effect of garlic powder extract on lipid content in normal and atherosclerotic human aortic cells. AB - In the present study, the mechanism of the in vitro effect of garlic powder extract (GPE) on lipid content of cultured human aortic cells was investigated. The addition of GPE abolished atherogenic blood serum-induced accumulation of free cholesterol, triglycerides, and cholesteryl esters in smooth muscle cells derived from uninvolved (normal) intima. In cells isolated from atherosclerotic plaque, GPE lowered these lipids. GPE inhibited lipid synthesis both in normal and atherosclerotic cells. It inhibited acyl-CoA:cholesterol acyltransferase activity that participates in the cholesteryl ester formation and stimulated cholesteryl ester hydrolase that degrades cholesteryl esters. This may explain the lipid reduction caused by GPE in atherosclerotic cells. GPE inhibited the uptake of modified low density lipoprotein and degradation of lipoprotein-derived cholesteryl esters, thus considerably reducing the intracellular accumulation of cholesteryl esters. This suggests the mechanism responsible for the prevention of lipid accumulation in aortic cells caused by atherogenic blood serum. PMID- 9358432 TI - Breast milk fatty acid composition: a comparative study between Hong Kong and Chongqing Chinese. AB - The fatty acids of milk samples obtained from 51 Hong Kong Chinese and 33 Chongqing Chinese (Si Chuan Province, China) were analyzed by gas-liquid chromatography. Compared with those of published data for Canadian and other Western countries, the Chinese milk from both Hong Kong and Chongqing contained higher levels of longer-chain polyunsaturated fatty acids, particularly docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6). In contrast, the content of trans fatty acids in the Chinese milk was lower compared with those for Canadian and other Western countries. Longitudinally, the concentrations of 22:6n-3 and 20:4n-6 gradually decreased when lactation progressed from colostrum (week 1) to mature (week 6). Over the same interval, linoleic acid (18:2n-6) remained unchanged in Chongqing Chinese but significantly increased in Hong Kong Chinese. Unlike 18:2n-6, linolenic acid (18:3n-3) increased in Chongqing Chinese but remained unchanged in Hong Kong Chinese throughout the study. The total milk fat also increased with the duration of lactation. In addition, the milk of Chongqing Chinese had higher total milk fat than that of Hong Kong Chinese and Canadians. The content of erucic acid (22:1n-9) increased with the progression of lactation in Chongqing Chinese, indicating that there was a switch in dietary consumption from fats of animal origin to rapeseed oil when lactation reached week 6. The present study showed that Hong Kong and Chongqing Chinese had a different fatty acid profile in many ways, which largely reflected a different dietary habit and life-style in these two places. PMID- 9358433 TI - Comparative hypocholesterolemic effects of six dietary oils in cholesterol-fed rats after long-term feeding. AB - Rats (8 wk of age) fed a conventional diet were shifted to diets containing 10% Oenothera biennis Linn oil (OBLO, linoleic acid + gamma-linolenic acid) from a wild plant, evening primrose oil (EPO, linoleic acid + gamma-linolenic acid) from a cultivated plant, bio-gamma-linolenic acid oil from mold (BIO, palmitic acid + oleic acid + linoleic acid + gamma-linolenic acid), safflower oil (linoleic acid), palm oil (PLO, palmitic acid + oleic acid + linoleic acid), or soybean oil (linoleic acid + alpha-linolenic acid) with 0.5% cholesterol for 13 wk. Though there were no significant differences in the food intake among the groups, the body weight gain of the OBLO group was significantly lower than that of the other groups except for the BIO and PLO groups, and that of the EPO and SBO groups were the highest among the groups. The liver weight of the OBLO group was significantly lower than that of other groups, and that of the PLO group was the highest among the groups. The serum total cholesterol and very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) + low density lipoprotein (LDL) cholesterol concentrations of the OBLO and EPO groups were consistently lower than those in the other groups. However, those of the BIO group were higher than those in the OBLO and EPO groups. The liver cholesterol concentration of the PLO group was the highest among all groups except for the EPO group. The fecal neutral sterol and bile acid extraction of the BIO group tended to increase compared to the other groups. The results of this study demonstrate that OBLO and EPO inhibit the increasing of serum total cholesterol and VLDL + IDL + LDL-cholesterol concentrations in the presence of excess cholesterol in the diet compared with the other dietary oils. PMID- 9358435 TI - Molecular speciation of fish sperm phospholipids: large amounts of dipolyunsaturated phosphatidylserine. AB - The molecular species compositions of the main diacyl phosphoglyceride classes and ether-linked subclasses from sperm of three species of fish, sea bass Dicentrarchus labrax, Atlantic salmon Salmo salar and Chinook salmon Onchorhynchus tsawytscha, were determined. The phospholipids from sperm were highly unsaturated, dipolyunsaturated fatty acid (diPUFA) molecular species comprised 64.6 to 71.8% of phosphatidylserine (PS), 10.1 to 17.4% of phosphatidylethanolamine (PE), and 3.3 to 10.1% of phosphatidylcholine (PC). In sea bass sperm, di22:6n-3 phospholipid was the predominant diPUFA molecular species, but in both salmon species 22:5n-3/22:6n-3 was also a major constituent of PS. Phospholipids containing 22:6n-3 dominated in sea bass sperm with 16:0/22:6n-3 as a major component of PC and PE, and 18:0/22:6n-3 of PE and PS in addition to di22:6n-3 in the latter two classes. In contrast, both salmon species contained much more 20:5n-3 and less 22:6n-3 so that saturated/20:5n-3 and monounsaturated/20:5n-3 molecular species were more abundant than the corresponding molecules containing 22:6n-3. Ether-linked lipids comprised 11.3 36.3% of choline and ethanolamine phosphoglycerides in each fish species. Molecular species containing 22:6n-3 were the major components of 1-O-alkyl-2 acyl-glycerophosphocholine, especially 16:0a/22:6n-3 in sea bass and 18:1a/22:6n 3 in the two salmon species, while in 1-O-alk-1'-enyl-2-acyl glycerophosphoethanolamine, 16:0a/22:6n-3 was the major component in both salmon and 18:0a/22:6n-3 in sea bass with 18:1a/22:6n-3 abundant in all three species. In Atlantic salmon 1-O-alkyl-2-acyl-glycerophosphoethanolamine comprised 24.6% of ethanolamine glycerophospholipids which were predominantly 16:0a/22:6n-3 and 18:1a/22:6n-3. Phosphatidylinositol from sperm was dominated by stearoyl/C20 PUFA molecular species, in sea bass overwhelmingly 18:0/20:4n-6, while in both salmon species 18:0/20:4n-6 and 18:0/20:5n-3 were equally abundant. PMID- 9358436 TI - Lipids and buoyancy in Southern ocean pteropods. AB - The lipids of Clione limacina, a Southern Ocean pteropod (order Gymnosomata), contain 28% diacylglyceryl ether (DAGE) (as percentage of total lipid) whereas the pteropod Limacina helicina (order Thecosomata) lacks DAGE. The alkyl glyceryl ether diols (1-O-alkyl glycerols, GE) of Clione DAGE are dominated by 16:0 (60%) and 15:0 (21%), in contrast with deep-sea shark liver DAGE, which is dominated by 18:1 GE. The fatty acid profiles of Clione and Limacina are similar (28-32% polyunsaturated, 26-34% monounsaturated) as are the sterols, which include 24 methylenecholesterol, transdehydrocholesterol, cholesterol, and desmosterol. This finding probably reflects the fact that Limacina is the major food source for Clione. Spongiobranchaea australis, another Southern Ocean pteropod (order Gymnosomata), has 0.9-1.7% DAGE, but has less lipid (3.3-4.8 mg/g lipid, wet weight) than Clione (50.8 mg/g lipid, wet weight). We propose a buoyancy role for DAGE in Clione since Limacina has bubbles for flotation which Clione lack; DAGE provides 23% more uplift than triacylglycerol at a concentration of 1.025 g/mL seawater. PMID- 9358434 TI - Low-dose eicosapentaenoic or docosahexaenoic acid administration modifies fatty acid composition and does not affect susceptibility to oxidative stress in rat erythrocytes and tissues. AB - In view of the promising future for use of n-3 polyunsaturated fatty acids (PUFA) in the prevention of cancer and cardiovascular diseases, it is necessary to ensure that their consumption does not result in detrimental oxidative effects. The aim of the present work was to test a hypothesis that low doses of eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) do not induce harmful modifications of oxidative cell metabolism, as modifications of membrane fatty acid composition occur. Wistar rats received by gavage oleic acid, EPA, or DHA (360 mg/kg body weight/day) for a period of 1 or 4 wk. Fatty acid composition and alpha-tocopherol content were determined for plasma, red blood cell (RBC) membranes, and liver, kidney, lung, and heart microsomal membranes. Susceptibility to oxidative stress induced by tert-butylhydroperoxide was measured in RBC. EPA treatment increased EPA and docosapentaenoic acid (DPA) content in plasma and in all the membranes studied. DHA treatment mainly increased DHA content. Both treatments decreased arachidonic acid content and n 6/n-3 PUFA ratio in the membranes, without modifying the Unsaturation Index. No changes in tissue alpha-tocopherol content and in RBC susceptibility to oxidative stress were induced by either EPA or DHA treatment. The data suggest that EPA and DHA treatments can substantially modify membrane fatty acids, without increasing susceptibility to oxidative stress, when administered at low doses. This opens the possibility for use of low doses of n-3 PUFA for chemoprevention without risk of detrimental secondary effects. PMID- 9358437 TI - Sulfoquinovosyl diacylglycerols from the alga Heterosigma carterae. AB - An extract of the chloromonad Heterosigma carterae (Raphidophyceae), cultivated in natural seawater, contained a complex mixture of sulfoquinovosyl diacylglycerols. Palmitoyl (16:0), three isomers of hexadecenoyl (16:1 cis delta 9, delta 11, delta 13), and eicosapentenoyl (20:5) were found to be the main fatty acyl substituents. Exact double-bond sites were determined by mass spectrometry analysis of the corresponding nicotinyl derivatives. Four major sulfoquinovosyl diacylglycerol components were partially purified and identified as 1-4 by interpretation of their nuclear magnetic resonance and mass spectral data. In addition, complete analysis of the H. carterae sulfoquinovosyl diacylglycerols was performed using high-performance liquid chromatography combined with electrospray tandem mass spectrometry. PMID- 9358439 TI - Oxidation reactions of acetylenic fatty esters with selenium dioxide/tert-butyl hydroperoxide. AB - Reaction of methyl undec-10-ynoate (1) with selenium dioxide/tert-butyl hydroperoxide (TBHP) in aqueous dioxane gave methyl 9-oxo-undec-10-ynoate (2, 9%) and 9-hydroxy-undec-10-ynoate (3, 60%), while methyl octadec-9-ynoate (4) yielded mixtures of positional isomers of mono-keto (viz. methyl 8-oxo- and 11-oxo octadec-9-ynoate, 5, 5%), hydroxy-keto (viz. methyl 8-hydroxy-11-oxo- and 11 hydroxy-8-oxo-octadec-9-ynoate, 6, 10%), and dihydroxy (viz. methyl 8,11 dihydroxy-octadec-9-ynoate, 7, 24%) derivatives. Similar treatment of a conjugated diacetylenic fatty ester (methyl octadeca-6,8-diynoate, 8) furnished a mixture of methyl 5-oxo- and 10-oxo-octadeca-6,8-diynoate (9, 12%) and a complex mixture of very polar products. Reaction of methyl octadec-11E-en-9-ynoate (methyl santalbate) (10) with selenium dioxide/TBHP in aqueous dioxane gave exclusively a mixture of regiospecific products, viz. methyl 8-oxo-octadec-11(E)Z en-9-ynoate (11, 6%) and methyl 8-hydroxy-octadec-11E-en-9-ynoate (12, 70%). The structures of the various products were determined by a combination of spectroscopic and mass spectral analyses. PMID- 9358440 TI - Lateral geniculate activations can be detected using intersubject averaging and fMRI. AB - Applications of fMRI in functional brain imaging are mainly confined to single subject designs, prohibiting the assessment of subject or group by condition interactions (i.e., differential activations) or areas of conjoint activation. In this paper a framework for fMRI group designs, using statistical parametric mapping, is introduced. It is generally believed that intersubject averaging, which requires spatial normalization and smoothing, will decrease the effective spatial resolution of fMRI or its sensitivity. A subcortical activation of the lateral geniculate nucleus (LGN) was therefore chosen to demonstrate the feasibility and power of intersubject averaging in the context of fMRI. Seven volunteers were studied, while looking at a blank screen or radially moving dots. LGN activation was demonstrated in all single subject analyses and in the group analysis. PMID- 9358438 TI - Measurement of apolipoprotein B in various cell lines: correlation between intracellular levels and rates of secretion. AB - We have standardized simple but sensitive enzyme-linked immunoassays to understand a relationship between intracellular levels and secretion rates of apoB. The assays were based on commercially available antibodies and were specific to human apoB. A monoclonal antibody, 1D1, was immobilized on microtiter wells and incubated with different amounts of low density lipoproteins to obtain a standard curve. Conditioned media were added to other wells in parallel, and the amount of apoB was quantitated from a linear regression curve. To standardize conditions for the measurement of intracellular apoB, cells were homogenized and solubilized with different concentrations of taurocholate. We found that 0.5% taurocholate was sufficient to solubilize all the apoB in HepG2, Caco-2, and McA RH7777 cells. Next, a standard curve was prepared in the presence of taurocholate and used to determine intracellular levels of apoB in different cell lines. The intracellular levels (pmol/mg cell protein) and the rates of secretion (pmol/mg/h) of apoB100 were positively correlated (r2 = 0.81, P = 0.0009) in HepG2 cells. Furthermore, a positive correlation (r2 = 0.88, P < 0.0001) was found between intracellular and secreted apoB42 in stably transfected McA-RH7777 cells. In contrast, no correlation was observed for human apoB28 and apoB18 in stably transfected cells that were secreted either partially associated or completely unassociated with lipoproteins. These studies indicated that the rate of secretion of lipid-associated apoB, but not the lipid-free apoB, was tightly controlled. PMID- 9358443 TI - Low visibility of lactate in excised rat muscle using double quantum proton spectroscopy. AB - Lactate NMR visibility was investigated in excised rat muscle at 3 T by comparing the concentration determined in situ by double quantum (DQ) proton spectroscopy (150 ms effective echo time) to the concentration measured in vitro from perchloric acid extracts of the same muscle samples. After 1-2 h of ischemia, lactate NMR visibility was 32 +/- 3% (+/- SE, n = 9), and was only 21 +/- 1% (n = 6) after 10-12 h. Muscle lactate T2 was 140 +/- 11 ms and 184 +/- 6 ms, respectively. All potential mechanisms of DQ lactate signal attenuation (B0 and B1 inhomogeneity, DQ transverse relaxation, diffusion) were examined, and accounted for when necessary. A significant increase in lactate NMR visibility was demonstrated using a shorter effective echo time (79 ms) DQ editing sequence. These results are interpreted as reflecting muscle lactate compartmentation between a long T2 pool predominantly detected by DQ spectroscopy, and a short T2 pool virtually invisible with longer echo time NMR techniques. PMID- 9358442 TI - An in vivo model for functional MRI in cat visual cortex. AB - A protocol is described for obtaining functional magnetic resonance images in anesthetized cat brain based on the blood oxygenation level dependent (BOLD) contrast mechanism. A visual stimulus was used, which consisted of a high contrast drifting grating, whose speed and spatial frequency was optimized for cat area 18 (V2). Experiments were conducted at 4.7 Tesla using a gradient echo EPI sequence with a 29-ms echo time, yielding signal changes of between 0.7% and 2% in area 18. PMID- 9358441 TI - Brain MRI with laser-polarized 129Xe. AB - The feasibility of brain MRI with laser-polarized 129Xe in a small animal model is demonstrated. Naturally abundant 129Xe is polarized and introduced into the lungs of Sprague-Dawley rats. Polarized xenon gas dissolves in the blood and is transported to the brain where it accumulates in brain tissue. Spectroscopic studies reveal a single, dominant, tissue-phase NMR resonance in the head at 194.5 ppm relative to the gas phase resonance. Images of 129Xe in the rat head were obtained with 98-microliter voxels by 2D chemical shift imaging and show that xenon is localized to the brain. This work establishes that nuclear polarization produced in the gas phases survives transport to the brain where it may be imaged. Increases in polarization and delivered volume of 129Xe will allow clinical measurements of regional cerebral blood flow. PMID- 9358444 TI - High relaxivity linear Gd(DTPA)-polymer conjugates: the role of hydrophobic interactions. AB - A series of linear copolymers of DTPA-class Gd3+ conjugates, linked by alpha, omega-alkyldiamides with a varying number (n) of methylenes separating the amide function, were synthesized. Surprisingly, their relaxivities at all fields increased with increasing n. At MRI fields and 35 degrees C, the relaxivities of the n = 10 and n = 12 polymers were unexpectedly high, similar to those of rigid dendrimer-based Gd3+ chelates. The magnetic field dependence of solvent proton 1/T1 was measured for aqueous urea-free and urea-containing polymer solutions. The results for urea-free solutions imply an increase of rigidity (required for high relaxivities) with increasing n, arising from hydrophobic interactions of the methylene groups with solvent. This hypothesis is supported by a large decrease in the relaxivities upon addition of urea, which is known to weaken hydrophobic interactions. The relaxivities are also independent of polymer concentration, indicating that the hydrophobic interactions are intramolecular. PMID- 9358445 TI - Dual-frequency, dual-quadrature, birdcage RF coil design with identical B1 pattern for sodium and proton imaging of the human brain at 1.5 T. AB - Rapid quantification of tissue metabolites in vivo by MRS or MRI can be achieved using dual-frequency RF coils with identical B1 field distributions at the observation frequencies of the metabolites and tissue water protons. Tissue sodium is used as an example for optimizing the dual-frequency, dual-quadrature RF coils for such measurements in humans. In the setting of sodium imaging, the challenge of dual-quadrature birdcage configurations is to decouple the sodium and proton channels because the fourth harmonic of the sodium frequency is very close to the proton frequency. A generalizable method for effectively decoupling these two RF frequencies is presented in this paper. The method is demonstrated with the design of an EPI compatible, dual-quadrature, double-tuned, 23Na/1H birdcage coil. The performance of the RF probe is reported at 1.5 Tesla in terms of signal-to-noise ratio, B1 homogeneity and image quality. PMID- 9358446 TI - Theory of the quadrature elliptic birdcage coil. AB - This paper presents the theory of the quadrature birdcage coil wound on an elliptic cylindrical former. A conformal transformation of the ellipse to a circular geometry is used to derive the optimal sampling of the continuous surface current distribution to produce uniform magnetic fields within an elliptic cylinder. The analysis is rigorous for ellipses of any aspect ratio and shows how to produce quadrature operation of the elliptic birdcage with a conventional hybrid combiner. Insight gained from the transformation is also used to analyze field homogeneity, find the optimal RF shield shape, and specify component values to produce the correct current distribution in practice. Measurements and images from a 16-leg elliptic birdcage coil at both low and high frequencies show good quadrature performance, homogeneity, and sensitivity. PMID- 9358447 TI - Measurement of the point spread function in MRI using constant time imaging. AB - The point spread function is a fundamental property of magnetic resonance imaging methods that affects image quality and spatial resolution. The point spread function is difficult to measure precisely in magnetic resonance even with the use of carefully designed phantoms, and it is difficult to calculate this function for complex sequences such as echo-planar imaging. This report describes a method that measures the point spread function with high spatial resolution at each pixel in samples of uniform intensity distribution. This method uses additional phase encoding gradients before the echo-planar acquisition that are constant in length but vary in amplitude. The additional gradients are applied to image the contents within each individual voxel. This method has been used to measure the point spread function for echo-planar imaging to demonstrate the effects of limited k-space sampling and transverse relaxation, as well as the effects of object motion. By considering the displacement of the point spread function, local distortions due to susceptibility and chemical shift effects have been quantified and corrected. The method allows rapid assessment of the point spread function in echo-planar imaging, in vivo, and may also be applied to other rapid imaging sequences that can be modified to include these additional phase encoding gradients. PMID- 9358448 TI - Isotropic diffusion-weighted and spiral-navigated interleaved EPI for routine imaging of acute stroke. AB - An interleaved echo-planar imaging (EPI) technique is presented for the rapid acquisition of isotropic diffusion-weighted images of stroke patients. Sixteen isotropic diffusion-weighted images at three b values are acquired in less than 3 min. A spiral navigator echo is used to measure the constant and linear phase shifts across the head in both the x and y directions which result from motion during the isotropic diffusion- sensitizing gradients. The measured k-space errors are corrected during a gridding reconstruction. The gridding kernel has a constant width in kx and a variable width in ky which eliminates variable data density ghosts. The resulting isotropic diffusion-weighted images have excellent lesion-to-normal brain contrast, very good spatial resolution, and little sensitivity to susceptibility effects in the base of the brain. Examples of diffusion-weighted images and ADC maps from several stroke patients are shown. PMID- 9358449 TI - Axonal stimulation under MRI magnetic field z gradients: a modeling study. AB - The stimulation of axons under MRI magnetic fields is analyzed solving the cable equation along the axons in the presence of magnetic field z gradients. Axons are represented using a one-dimensional compartmental cable model including the kinetics of mammalian myelinated fibers. Computer simulations of the model were performed for sinusoidal and trapezoidal fields. Several axon and field parameters were tested to determine the threshold values for axonal stimulation of the magnetic field, induced electric field and time derivative of the magnetic field. The results indicate that 1) threshold for stimulation is lowest for the largest diameter axons terminating in the region where the magnetic field is maximum, 2) trapezoidal waveforms can be optimized to allow better sub-threshold resolution than sinusoidal waveforms, and 3) the induced electric field is better than the magnetic field and time derivative of the magnetic field as indicators of stimulation threshold. PMID- 9358450 TI - T2 accuracy on a whole-body imager. AB - MR oximetry requires a T2 measurement that is accurate within 5% in vivo. Simple methods are susceptible to signal loss and tend to underestimate T2. Current methods utilize RF pulses or RF cycling patterns that prevent signal loss at each data acquisition. However, using these methods with imperfect pulses, T2 tends to be overestimated due to temporary storage of the magnetization along the longitudinal axis where it decays more slowly with a time constant T1 > T2. To reduce the T1 dependence while preventing signal loss, we utilize simple 90x180y90x composite pulses and good RF cycling patterns. These trains are critical for T2 accuracy over typical ranges of RF and static field inhomogeneities and refocusing intervals. T1 signal decay during each 90x180y90x pulse must be accounted for to yield accuracy within 5% when the pulse-width is 10% or more of the refocusing interval. A simple correction scheme compensates for this T1-related error effectively. PMID- 9358451 TI - Study of the metabolism of choline and phosphatidylcholine in tumors in vivo using phosphonium-choline. AB - The results of an initial study on the feasibility of using the phosphonium analog of choline to follow the metabolism of phosphatidylcholine in tumors in vivo using 31P NMR are reported. C3H/He mice bearing a mammary carcinoma tumor on the foot pad were fed a choline-free diet supplemented with the phosphonium analog of choline. Metabolites of this compound, including the phosphonium analogs of phosphatidylcholine, phosphocholine, glycerophosphocholine, and betaine were observed noninvasively in vivo in tumors by 31P NMR after 2-3 weeks of feeding. Clearance of these phosphonium-labeled metabolites from tumors was measured after a change to a choline-containing diet. Significant decreases were seen in the levels of the analogs of betaine (P < 0.003) and phosphatidylcholine (P < 0.004) by Day 4. A significant increase in the level of authentic phosphocholine (P < 0.003) occurred over the same time period. PMID- 9358452 TI - Noninvasive comprehensive characterization of renal artery stenosis by combination of STAR angiography and EPISTAR perfusion imaging. AB - In a small fraction of patients with hypertension, the cause is stenosis of the renal artery. To date, there is no established noninvasive screening technique available to identify this population, for whom treatment with a surgical procedure or percutaneous transluminal angioplasty is often possible. In this study, the sensitivity and specificity of STAR angiography and EPISTAR perfusion imaging in characterizing renal artery stenosis were evaluated in an animal model. Qualitatively, STAR provided projection angiograms that were comparably sensitive to x-ray angiograms but obtained noninvasively. The sensitivity for detecting the stenosis was 100%. The specificity varied according to the definition of the threshold for significance, which varied 78-94%, depending on whether > 70% or > 50% diameter reduction was considered. Improvements in specificity will depend upon use of shorter echo times and higher spatial resolution. Our preliminary data indicate that EPISTAR provides sensitivity and specificity of 100% for characterizing stenosis with diameter reductions of > 70%. PMID- 9358453 TI - A simplified sequence for observing deoxymyoglobin signals in vivo: myoglobin excitation with dynamic unexcitation and saturation of water and fat (MEDUSA). AB - This paper describes a new, simplified pulse sequence for observing NMR signals from deoxymyoglobin in vivo. Paramagnetically shifted resonances from deoxymyoglobin can be exploited to noninvasively calculate intracellular oxygen tension in striated muscle. However, special sequences are required to observe these weak signals against the larger water and fat signals encountered in vivo. The pulse sequence described here, which is based on inversion recovery sequences, efficiently suppresses both water and fat resonances and can be implemented with short repetition rates. Moreover, it is perfectly suited for studies with surface coils, where RF inhomogeneities render other popular suppression sequences ineffective. PMID- 9358454 TI - Performance of an elliptical centric view order for signal enhancement and motion artifact suppression in breath-hold three-dimensional gradient echo imaging. AB - An elliptical centric phase-encoding (PE) order is applied to steady-state 3DFT imaging as performed during a single breath-hold or following contrast agent administration. In a set of simulation, phantom, and in vivo experiments, this truly centric PE order is shown to be both more resistant to breathing artifact and more capable of suppressing undesirable venous signals that can arise following peak arterial enhancement than a number of other centric PE techniques that are presently in use. Unlike other PE orders, the elliptical centric ordering changes with the relative dimensions of the two PE fields of view and is optimal based on increasing k-space radius. It thus creates a temporally diminishing importance to the PE order. The specific advantages of such an acquisition are demonstrated. PMID- 9358455 TI - Myocardial function in infarcted and remote regions early after infarction in man: assessment by magnetic resonance tagging and strain analysis. AB - Early after infarction in the perfusion bed of the left anterior descending coronary artery, cine MRI with spatial modulation of magnetization (SPAMM) tagging (7-mm grid) was used for short- and long-axis cardiac imaging. Two dimensional strain analysis of triangular finite elements was performed between end-diastole and end-systole. Patients (n = 10) were compared with age-matched healthy subjects (n = 8). The anteroseptal region at midventricular level was considered representative for "infarcted" and the posterolateral region at basal level was considered "remote". The left ventricular end-diastolic volume index was larger in the patients (69 +/- 15 ml/m2 versus 56 +/- 4 ml/m2, P < 0.05). Short-axis images showed in the infarcted region a decrease of first principal strain (greatest systolic lengthening: 1.10 +/- .06 versus 1.27 +/- 0.04, P < 0.0001), and in the remote region an increase (1.48 +/- 0.11 versus 1.36 +/- 0.07, P < 0.025). The lateral and inferior ventricular regions at mid- and basal levels were found to function normally. Long-axis images yielded similar results. Early after infarction, regions with dysfunction, normal function, and hyperfunction can be delineated with MR tagging. The compensatory increased contraction in the remote region is possibly triggered by the Frank-Starling mechanism. PMID- 9358456 TI - Statistical methods in functional magnetic resonance imaging with respect to nonstationary time-series: auditory cortex activity. AB - In awake animal and human auditory cortices, it is a common experience with electrophysiological and suitable imaging methods for responses to steady stimulation to be strongly state-dependent and to exhibit nonstationarities, even over short periods of observation. If such nonstationary behavior is also reflected by hemodynamic responses in the human auditory cortex, conventional methods of analysis of fMRI data, although applicable for instance to largely stationary responses in visual and other cortices, may be misleading in attempts to parcellate auditory cortex into fields and to demonstrate functional maps. Time-Windows, described in this article as a convenient tool for the detection and analysis of time-variant brain activities, solves some of these problems. Time-Windows demonstrates that activity is evoked reliably in three separate territories of human auditory cortex, parts of which may show nonstationary behavior, depending on the auditory stimuli and tasks. PMID- 9358457 TI - Evaluation of triple quantum-filtered 23Na NMR spectroscopy in the in situ rat liver. AB - Triple quantum (TQ)-filtered 23Na NMR spectroscopy and the shift reagent, TmDOTP5 , have been used to evaluate the contributions of intra- (Na+i) and extracellular (Na+e) sodium to the TQ-filtered signal in the rat liver, in situ. Na+e contributed significantly to the total TQ-filtered signal in live animals, and the intensity of this signal did not change postmortem. The TQ-filtered Na+i signal increased by approximately 380% over a period of 1 h postmortem, whereas the single quantum (SQ) Na+i increased by 90%. The constancy of the TQ-filtered Na+i signal indicates that changes in total TQ-filtered 23Na signal intensity in liver (without a shift reagent) may accurately reflect changes in TQ-filtered Na+i signal intensity. The large percent increase in the TQ-filtered Na+i signal as compared to the SQ signal suggests that the fraction of Na+i interacting with macromolecules increases after death. PMID- 9358458 TI - Implementation and evaluation of frequency offset corrected inversion (FOCI) pulses on a clinical MR system. AB - FOCI pulses are a variant of hyperbolic secant pulses in which the RF amplitude, RF frequency, and gradient waveform are all modulated by the same function A(t). This increases the usable gradient amplitude without requiring a corresponding increase in RF amplitude. In this paper the implementation and inversion profiles of FOCI pulses on a clinical MR system are described, showing improved slice definition and chemical-shift offset behavior. Their adiabatic behavior is confirmed, and their use with an ISIS sequence for localized MRS is illustrated. Finally the effects of waveform digitization are considered, and the implications for SAR and dB/dt are discussed. PMID- 9358459 TI - Improvements on an in vivo automatic shimming method [FASTERMAP]. AB - Improvements on a localized, automatic shimming method described by Gruetter and Boesch (J. Magn. Reson. 96, 323-334 (1992)) and Gruetter (Magn. Reson. Med. 29, 804-811 (1993)) are presented. A spin-echo sequence employing a double sech refocusing scheme is used to acquire a field map along linear projections that improves the signal-to-noise ratio by at least a factor of two over the stimulated echo sequence previously used. To further improve the reliability of shim adjustments, a variance-weighted polynomial regression analysis is performed. This also extends the scope of application of this technique to global shimming. Localized 1H spectra of human brain shimmed by this method are presented. PMID- 9358460 TI - Assessment of quantitative artificial neural network analysis in a metabolically dynamic ex vivo 31P NMR pig liver study. AB - Quantitative artificial neural network analysis for 1550 ex vivo 31P nuclear magnetic resonance spectra from hypothermically reperfused pig livers was assessed. These spectra show wide ranges of metabolite concentrations and have been analyzed using metabolite prior knowledge based lineshape fitting analysis which had proved robust in its biochemical interpretation. This finding provided a good opportunity to assess the performance of artificial neural network analysis in a biochemically complex situation. The results showed high correlations (0.865 < or = R < or = 0.992) between the lineshape fitting and artificial neural network analysis for the metabolite values, and the artificial neural network analysis was able to fully represent the trends in the metabolic fluctuations during the experiments. PMID- 9358461 TI - Temperature mapping using the water proton chemical shift: a chemical shift selective phase mapping method. AB - A proton-chemical-shift-based temperature imaging method, called chemical shift selective phase mapping, is proposed. The technique uses frequency-selective suppression to provide frequency selectivity to the phase mapping method. Separate imaging of the phase distributions of the water and nonwater signals reduced the error due to the presence of a nonwater signal in measuring the water proton chemical shift change in two-component samples. Imaging of the phase difference between water and oil yielded an internally referenced water proton chemical shift measurement to visualize the temperature change distribution, which was unaffected by motion-induced susceptibility changes. PMID- 9358463 TI - Planar gradient coil design by scaling the spatial frequencies of minimum inductance current density. AB - Gradient coil inductance has been remarkably reduced by the minimum-inductance design technique, which minimizes the magnetic energy stored by the gradient coil. The planar gradient coil designed by this technique, however, often has poor magnetic field linearity. Scaling the spatial frequencies of the current density function derived by this method, the magnetic field linearity of the planar gradient coil can be greatly improved with a small sacrifice of gradient coil inductance. A figure of merit of the planar gradient coil has been found to be improved by scaling the spatial frequencies. PMID- 9358462 TI - Signal-to-noise measurements in magnitude images from NMR phased arrays. AB - A method is proposed to estimate signal-to-noise ratio (SNR) values in phased array magnitude images, based on a region-of-interest (ROI) analysis. It is shown that the SNR can be found by correcting the measured signal intensity for the noise bias effects and by evaluating the noise variance as the mean square value of all the pixel intensities in a chosen background ROI, divided by twice the number of receivers used. Estimated SNR values are shown to vary spatially within a bound of 20% with respect to the true SNR values as a result of noise correlations between receivers. PMID- 9358465 TI - Two monoclonal antibodies better than one? PMID- 9358464 TI - Soluble complement component tackles reperfusion injury. PMID- 9358466 TI - Stimulating drugs in Alzheimer's disease. PMID- 9358467 TI - Novel drugs and strategies for gastrointestinal disorders. PMID- 9358469 TI - Memories are made of this: the genetic basis of memory. AB - Total amnesia is rare, but we face an 'epidemic' of memory loss. At present there are around 18 million people worldwide with Alzheimer's disease, and this figure is predicted to double in the next 25 years. While traditional clinical and experimental studies have elucidated much about the basic processes of memory and learning, modern genetic techniques. Only time will tell whether this knowledge will yield preventive or curative therapy for memory loss. PMID- 9358468 TI - Allergic contact dermatitis: understanding the immune response and potential for targeted therapy using cytokines. AB - Allergic contact dermatitis is the most common job-related disease of the western world. The only available treatments are avoidance of contact with the allergen and the use of potent corticosteroids. Recently, the role of cytokines in the pathogenesis of this disease has been studied and, besides defining the key molecules and basic cellular immune responses responsible for disease development, these studies might help to develop new therapeutic strategies to target cytokines and thereby try to alter or abrogate ongoing immune reactions. PMID- 9358470 TI - Idiotypic vaccination in B-cell malignancies. AB - Immunoglobulins contain unique portions, collectively termed idiotypes, that can be recognized by the immune system. Idiotypes expressed by tumor cells in B-cell malignancies can be regarded as tumor-specific antigens and targets for vaccine immunotherapy. Haptens and adjuvants, including cytokines, have been used in several animal models to increase idiotype immunogenicity and establish protective anti-idiotype immunity. These results have been extended by the use of DNA technology, and this has led to the development of a new generation of immunogens, namely fusion proteins and naked-DNA vaccines. The central role of antigen-presenting cells as initiators of anti-idiotype immune responses has also been recognized. Guided by the experimental data, idiotypic vaccination has come into medical use in patients with lymphoma and multiple myeloma. PMID- 9358471 TI - Leukoplakia: molecular understanding of pre-malignant lesions and implications for clinical management. AB - Oral leukoplakia is a white patch or plaque of the oral mucosa that cannot be characterized clinically or pathologically as any other diagnosable disease. Although oral leukoplakia is a well-known oral premalignant lesion with a high risk for development of oral cancer, little is known about its genetic basis. The development of oral cancer is a multistep process and it is believed that multiple genetic alterations are involved. Understanding this underlying genetic basis of oral cancer development will help us design better diagnosis and treatment plans in the cancer clinic. This review discusses recent progress in the identification of genetic and cytogenetic alterations in oral pre-malignant lesions and their potential clinical applications. PMID- 9358472 TI - Cell adhesion: a new target for therapy. AB - Intercellular adhesive events are involved in a wide range of biological processes, including pattern formation and morphogenesis during development, immune responses, leukocyte recirculation, wound repair, tumour growth and metastasis. In the multicellular state, signals from cell adhesion molecules, along with those from growth factor and cytokine receptors, provide a range of information to the cell that is integrated to yield a final message, perhaps to maintain the cell cycle (if it is a stem cell) or follow a path towards terminal differentiation. Aberrant cell adhesion plays a key role in many developmental defects, acute and chronic inflammatory disease and cancer. PMID- 9358474 TI - The ICD--progress, prospects, and problems. PMID- 9358473 TI - Epidermolysis bullosa: a spectrum of clinical phenotypes explained by molecular heterogeneity. AB - Great progress has recently been made in understanding the molecular basis of various heritable skin diseases. A prototype of such conditions is epidermolysis bullosa (EB), a heterogeneous group of mechano-bullous disorders characterized by fragility of the skin and other specialized epithelia. Blistering of the skin in EB results either from fragility of epidermal cells or from defective attachment of the epidermis to the underlying dermis, because of genetic lesions within molecules of the basement-membrane zone at the dermal-epidermal junction. Distinct mutations have been discovered in ten different genes encoding the structural components within this layer. The combinations and the types of mutations, as well as their positions in the altered gene products, collectively reflect the phenotypic variability observed in this group of heritable skin diseases. PMID- 9358475 TI - Role of sympathovagal balance in the initiation of idiopathic ventricular tachycardia originating from right ventricular outflow tract. AB - VT originating from the right ventricular outflow tract (RVOT) is prone to occur when sympathetic nervous activity is increased. beta-Blockade is, therefore, effective in suppressing this VT. The purpose of this study was to determine the role of sympathovagal balance assessed by heart rate variability (HRV) in the spontaneous initiation of repetitive premature ventricular contractions (PVCs) and VT (five or more consecutive PVCs) arising from RVOT in seven patients without structural heart diseases. Frequency-domain measures of HRV were determined by analyzing 24-hour Holter electrocardiographic recording with the maximum entropy method over a 1,280-second period immediately before the onset of 35 single PVCs, 26 episodes of 2-4 consecutive PVCs, and 21 episodes of VT. High frequency component (HF: 0.15-0.40 Hz) was used as an index of parasympathetic activity, and the ratio of low frequency component (LF: 0.04-0.15 Hz) to HF (LF/HF ratio), as an index of sympathovagal balance. NN50(%), a time-domain variable of parasympathetic activity, was also determined. Mean RR interval and any measures of HRV did not change significantly before single PVCs. Mean RR interval shortened and HF decreased prior to repetitive PVCs and VT. The LF/HF ratio, however, increased only before the onset of VT. NN50(%) tended to decrease before repetitive PVCs and decreased significantly before VT. With propranolol (30-60 mg/day), frequency of repetitive PVCs was suppressed from 2,048 +/- 1,201 to 746 +/- 658/day and VT was totally abolished, but frequency of single PVCs did not change significantly. In conclusion, sympathetic predominance plays an important role in the initiation of repetitive PVCs and VT originating from RVOT in patients without structural heart diseases. PMID- 9358476 TI - Ventriculoatrial conduction capability and prevalence of 1:1 retrograde conduction during inducible sustained monomorphic ventricular tachycardia in 305 implantable cardioverter defibrillator recipients. AB - Despite the advent of dual chamber ICDs, differentiation of VT (SMVT) with 1:1 VA conduction will remain a challenge. In this study, VA conduction capability and prevalence of inducible sustained monomorphic (SM) VT with 1:1 VA conduction was assessed in 305 ICD recipients. SMVT with a mean cycle length (CL) of 304 +/- 61 ms was induced in 161 (53%) patients. Twenty-six percent of the patients maintained 1:1 VA conduction to CL < or = 400 ms during incremental ventricular pacing, regardless of presenting tachyarrhythmia or presence of inducible SMVT. Among ten patients who had inducible SMVT with possible 1:1 VA conduction (based on SMVT CL comparable to the shortest CL associated with 1:1 retrograde conduction during ventricular pacing), all seven with available intracardiac tracings had documented 1:1 VA conduction during the induced SMVT--representing 4.4% of the patients with inducible SMVT (95% CI 1.2%-7.6%), and 2.3% of the entire ICD cohort (95% CI 0.6%-4.0%). We conclude that about one-fifth of ICD recipients possess 1:1 VA conduction to CL < or = 400 ms and that inducible SMVT with 1:1 VA conduction can be demonstrated in a small but nonnegligible proportion of ICD recipients. These data are relevant to the design of tachyarrhythmia-discrimination algorithms for dual chamber ICDs. PMID- 9358478 TI - Do permanent pacemakers need an insulative coating? Results of a prospective randomized double-blind study. AB - During conventional manufacturing of implanted pulse generators (IPGs), an insulative coating is often applied to prevent local muscle stimulation and myopotential sensing in unipolar pacing. This can limit the orientation of the IPG into its pocket, be a potential source of muscle stimulation via coating scratches, and result in an increase in IPG production costs. We hypothesized that advances in the design and construction of current IPGs and leads obviates the need for an insulative coating of the IPG. Using a double-blind prospective randomized design, 39 patients were implanted with either coated or uncoated otherwise identical IPGs (19 dual, 20 single chamber). All testing was done in unipolar and bipolar mode in both channels. A strength-duration curve for muscle stimulation was constructed for all patients with muscle stimulation. Myopotential sensing was established during isometric exercise. At 6-month follow up when tested in unipolar mode, 3 of 15 (20%) patients with coated IPGs and 3 of 20 (15%) with uncoated IPGs had muscle stimulation at 5.0 V/1.5 ms or lower (P = NS). No patients in either population had muscle stimulation at their normally programmed output. Myopotential sensing occurred in all patients in unipolar mode at a mean of 2.29 +/- 1.3 mV and 2.73 +/- 1.14 mV for coated versus uncoated, respectively (P = NS). The statistical power of these negative observations was 80%. An insulative coating for pacemakers does not appear to alter sensing performance or cause a significant difference in the occurrence or characteristics of muscle stimulation. PMID- 9358477 TI - Are double potentials markers of a specific zone of the atrioventricular junction in the isolated rabbit heart? AB - A study is made of the characteristics of the atrial potentials recorded in the Koch triangle and its proximity, their variations on modifying the site of cardiac pacing, and their usefulness as markers of a distinct zone of the AV junction. In 12 isolated and perfused rabbit heart preparations an analysis was made of the endocardial atrial electrograms recorded with a multiple electrode positioned in the AV junction. The electrograms were obtained during spontaneous rhythm and on pacing at the crista terminalis (CT), interatrial septum (IAS), left atrium, and right ventricle. Double potentials were frequently obtained. On pacing at the CT, high-low double potentials (DP [H-L]) were more frequent (P < 0.05) in the low CT (11% +/- 4% of the electrodes) and posterior zone of the Koch triangle (6% +/- 5%), than in the IAS (1% +/- 2%) and anterior zone of the Koch triangle (2% +/- 3%). A similar tendency was observed either on pacing at the left atrium or during spontaneous rhythm. During pacing at the IAS the percentages of low-high double potentials (DP (L-H]) were significantly higher (P < 0.05) in the low CT (7% +/- 6%). DP (H-L) were of low sensitivity in indicating a given zone; maximum sensitivity was 61% in the low CT when pacing at the CT. DP (L-H) proved even less sensitive in indicating a given zone, though their specificity was greater in the low CT (91%) during pacing at the IAS. The specific zones in which the highest percentages of DP (H-L) or DP (L-H) are obtained depend on the site of cardiac pacing. On pacing at the IAS, DP (L-H) are more specific of the low CT. During pacing at both the CT and at the IAS, DP (H L) sensitivity in indicating a given zone is low. PMID- 9358479 TI - Permanent atrial pacing in cardiac transplant patients. AB - Thirteen out of 223 consecutive cardiac transplant patients required permanent pacemaker implantation; 11 for sinus node dysfunction and 2 for complete AV block. Patients with sinus node dysfunction were considered for AAIR mode pacemakers if they had intact AV conduction defined as a Wenckebach point of > 120 beats/min. Ten of 11 patients with sinus node dysfunction had a single atrial lead placed. Atrial lead placement was most easily accomplished with a straight, active fixation lead and the use of manually curved stylets to find an optimal position in the donor atrium, although active fixation leads with a preformed atrial J were used as well. Two leads dislodged requiring revision. In contrast, only 1 of 250 atrial leads implanted in nontransplanted hearts dislodged (P < 0.0001). Transvenous endomyocardial biopsies have not caused atrial lead dislodgment. Most transplant recipients requiring permanent pacing have intact AV nodal function and require only atrial pacing. Atrial lead dislodgment requiring lead revision occurs more frequently in heart transplant recipients than in native hearts. Use of a straight active fixation lead with a manually formed curve in the stylet is useful in order to find the optimal position for pacing. PMID- 9358481 TI - A simple electrocardiographic algorithm for detecting ventricular tachycardia. AB - The purpose of this study was to determine whether a simple ECG algorithm could be developed for predicting susceptibility to ventricular tachyarrhythmias (VT) as defined by sustained spontaneous or inducible VT. Two different QT dispersion algorithms were determined by the difference between the longest and shortest QT interval measured in three orthogonal leads (I, aVF, V1; QTD3), and at least 11 of 12 leads (QTD12) from the 12-lead ECG. These QT dispersion algorithms were investigated (with and without the QRS duration from the 12-lead ECG) and compared to the signal-averaged ECG (SAECG) in order to determine their sensitivity and specificity for detecting VT. Only patients who underwent SAECG and were referred for programmed electrical stimulation were included in this study. A positive SAECG was defined by filtered QRS duration > 114 ms, and/or low amplitude signal duration > 38 ms, and/or root mean square voltage in the last 40 ms of < 20 microV. Sixty patients were enrolled in this study with a mean age of 63 +/- 2 years. Fifty-five percent of the patients had coronary artery disease. A simple ECG algorithm consisting of the sum of QTD3 plus the QRS duration had a sensitivity and specificity of 90% and 63%, respectively, wheras the SAECG had a sensitivity and specificity of 60% and 63%, respectively (P = 0.022). We conclude that a simple ECG algorithm is more sensitive than the SAECG for predicting VT. This algorithm combines two easily measured variables obtained from the 12-lead ECG, and can easily be performed without expensive computer equipment. PMID- 9358480 TI - Is dispersion of ventricular repolarization rate dependent? AB - QT dispersion has been adopted as a new index for the noninvasive assessment of the inhomogeneity of repolarization and has been evaluated in several clinical studies as an index of arrhythmia propensity. In most of these studies, indices of dispersion of repolarization were rate corrected by the Bazett formula calculating QT dispersion as QT cmax-QT cmin or JT dispersion as JT cmax-JT cmin, implying that dispersion of repolarization also changes with heart rate. This study aimed to determine in the electrically paced isolated heart whether dispersion of ventricular repolarization is rate dependent. Multiple (5-7) monophasic action potentials (MAPs) were recorded simultaneously from the epicardium and endocardium of both ventricles in 18 isolated Langendorff-perfused rabbit hearts. Hearts were paced from a right ventricular site at basic cycle lengths (CL) between 1,200 and 300 ms in 100-ms decrements. Action potential duration was measured at 90% repolarization (APD90), and recovery time (RT) was defined as the sum of APD90 and activation time in each of the simultaneous MAP recordings. The dispersion of APD90 and RT, respectively, were calculated as the maximal difference among all recordings. APD90 and RT shortened continuously throughout the range of paced steady-state CLs from 1,200 to 300 ms. APD90 was 197.6 +/- 6.1 ms at a CL of 1,200 ms and decreased to 148.5 +/- 2.5 ms at a CL of 300 ms (P < 0.0001). RT was 228.2 +/- 6.2 ms at a CL of 1,000 ms and decreased to 175.9 +/- 2.9 at a CL of 300 ms (P < 0.0001). In contrast, dispersion of APD90 and RT did not change significantly. Dispersion of APD90 was 24.8 +/- 2.3 ms at a CL of 1,200 ms, 26.1 +/- 1.9 msec at a CL of 1,000 ms, and 21.6 +/- 2.1 at a CL of 300 ms (NS). Dispersion of RT was 29.7 +/- 3.4 ms at a CL of 1,200 ms, 29.0 +/ 3.0 ms at a CL of 1,000 ms, and 32.7 +/- 3.2 ms at a CL of 300 ms (NS). In contrast to the duration of the QT interval, dispersion of ventricular repolarization does not change significantly with pacing induced changes in CL. Assuming that the rate-dependent behavior of action potential duration is similar between the rabbit and human heart, a rate correction of parameters of dispersion of repolarization is probably unnecessary. PMID- 9358482 TI - The effect of nonisodiametric design on the ease of extracting chronically implanted pacemaker leads. AB - Extracting permanently implanted transvenous pacemaker leads is often difficult because a fibrous sheath tends to trap the lead at various points along its course. Because many leads have bulbous or nonisodiametric portions, extraction may be rendered even more troublesome, because it is difficult to pull the larger portion through the narrow areas of the sheath. Furthermore, forceful extraction may have dire consequences, such as cardiac laceration. A study was undertaken in animals to evaluate the effect of lead isodiametricity on lead extraction. The results show that any increase in the diameter of the lead tip greatly reduces the ease of its removal. Consequently, leads designed to be isodiametric throughout their entire lengths will greatly enhance their removability. PMID- 9358483 TI - Hemodynamic deterioration following radiofrequency ablation of the atrioventricular conduction system. AB - Radiofrequency ablation of the atrioventricular conduction system (ACS) has become an established therapy for patients with drug refractory atrial fibrillation. We observed eight patients with hemodynamic deterioration after radiofrequency ablation of the atrioventricular conduction system. As we found hemodynamic deterioration related to worsening mitral regurgitation, we compared the clinical history, electrophysiological, and echocardiographic data from the patients with hemodynamic deterioration and worsening mitral regurgitation (group 1) to those without hemodynamic deterioration and stable mitral regurgitation after the procedure (group 2). Eight out of 108 patients (7.4%) undergoing ablation of the ACS deteriorated hemodynamically with acute pulmonary edema in three and congestive heart failure in five patients occurring at a mean of 3 and 8 weeks, respectively, after the procedure. Three of these patients were referred for mitral valve surgery. Two patients underwent ablation using a left-sided approach. A right-sided approach was used in five patients. In one patient, a left- and right-sided approach was used. Compared to group 2 patients, group 1 patients had significantly higher left ventricular end-diastolic diameters (64 +/ 6 mm vs 56 +/- 9 mm) at baseline despite similar fractional shortening (32% +/- 11% vs 34% +/- 13%), left ventricular end-systolic diameters (43 +/- 9 mm vs 36 +/- 7 mm) and degree of mitral regurgitation (1.4 +/- 1.1 vs 1.4 +/- 0.7) on echocardiographic analysis. Thus, hemodynamic deterioration together with progression of mitral regurgitation is a potential complication of ablation of the ACS (up to 7.4%). Patients with high left ventricular end-diastolic diameters and moderate mitral regurgitation at baseline seem prone to this complication. PMID- 9358484 TI - Atrial pacing leads following open heart surgery: active or passive fixation? AB - The right atrial appendage is often amputated at the time of cardiopulmonary bypass. Because of concerns regarding lead displacement, use of active fixation atrial leads has been recommended in patients who require permanent atrial or dual chamber pacing after open heart surgery. We evaluated the acute and chronic performance of active and passive fixation atrial leads implanted at our institution between 1985 and 1993 in patients with previous open heart surgery. Of 78 consecutive patients, 38 had an active fixation atrial lead, 28 had a passive fixation steroid-eluting lead, and 12 had a passive fixation lead without steroid-eluting properties. At implantation, sensed P wave amplitudes were similar in the three groups, but lead impedance and threshold were significantly higher for active fixation leads compared to all passive fixation leads. During follow-up, atrial pacing thresholds were significantly higher, and sensed P wave amplitudes significantly lower, in the patients with active fixation leads compared to those with passive fixation leads. Loss of sensing occurred in 6 of 38 (16%) patients with active fixation leads and 1 of 40 (2.5%) patients with a passive fixation lead (P = 0.027). Atrial lead displacement occurred in two patients with active fixation leads and one with a passive fixation lead. Comparison with a parallel group of patients without previous open heart surgery demonstrated that atrial lead performance was similar in the two groups. We conclude that, when permanent atrial or dual chamber pacing is necessary in patients with prior open heart surgery, it is appropriate to implant a passive fixation atrial lead except on the infrequent occasions when a stable atrial position cannot be obtained. PMID- 9358485 TI - Internal atrial defibrillation: effect on sinus and atrioventricular nodal function and implanted cardiac pacemakers. AB - Internal atrial defibrillation (IAD) has been extensively evaluated for clinical efficacy but the need for concomitant demand pacing and the effect of IAD shocks on pacemaker function is not well studied. We prospectively evaluated: (1) the incidence of bradycardia as a result of IAD shocks; and (2) effect of these shocks on functioning of implanted cardiac pacemakers. Consecutive consenting patients with atrial fibrillation (AF) requiring cardioversion or undergoing electrophysiological study were selected for IAD. IAD shocks were delivered using the right ventricle to right atrium (RV-RA), right ventricle to superior vena cava (RV-SVC), right atrium to axillary patch (RA-AX), and right atrium to left pulmonary artery or coronary sinus (RA-LPA/CS) lead configurations. Mean RR interval before and after the shocks and the time interval from shock delivery to first QRS complex were analyzed for unsuccessful and successful shocks. Pacing and sensing function was analyzed in patients with previously implanted pacemakers. Twenty-five patients, 18 men, mean age 67.9 +/- 10 years were included in the study. A total of 305 shocks (264 unsuccessful, 41 successful) were analyzed. For unsuccessful shocks the mean post-IAD shock RR interval (795 +/- 205 ms) and the time to first post-IAD shock QRS complex (970 +/- 438 ms) were both significantly greater than the pre-IAD shock RR interval (685 +/- 131 ms, P < 0.001). The increase in post-IAD shock RR interval and time to first post IAD shock QRS complex was seen with all four lead configurations used. With successful shocks the mean post-IAD shock sinus cycle length (1,105 +/- 450 ms) and time to first post-IAD shock QRS complex (1,126 +/- 443 ms) were both also significantly greater than the pre-IAD shock RR interval (766 +/- 172 ms). Nine patients (36%) had episodes of significant bradycardia after shock delivery. Shocks of up to 20 J using the RA-LPA/CS lead configuration did not affect pacemaker function. IAD can result in transient bradycardia related to sinus and atrioventricular nodal effects requiring backup ventricular pacing. Shocks can be safely delivered using RA-LPA or RA-CS lead configurations in patients with implanted bipolar cardiac pacemakers. PMID- 9358486 TI - A comparison of transvenous atrial defibrillation of acute and chronic atrial fibrillation and the effect of intravenous sotalol on human atrial defibrillation threshold. AB - The comparative efficacy and safety of transvenous defibrillation for acute and chronic AF and the effect of antiarrhythmic agents on this therapy have not been evaluated. Transvenous atrial defibrillation was performed in 25 patients with chronic AF and 13 patients with acute AF by delivering R wave synchronized, biphasic shocks between the right atrium and coronary sinus. The lowest energy and voltage resulting in successful defibrillation were considered to be atrial defibrillation threshold (ADFT). Intravenous sotalol (1.5 mg/kg) was then given over 15 minutes and ADFT was determined again. The mean ADFT was 1.5 J and 3.6 J for acute and chronic AF, respectively, and the threshold was highly reproducible. Sotalol reduced ADFT in patients with acute AF while the reduction in chronic AF group was not significant. There was no significant increase in creatinine kinase nor reduction in blood pressure, but prolonged pause after successful defibrillation required ventricular supporting pacing. We conclude that transvenous atrial defibrillation is a safe and effective means for defibrillating both acute and chronic AF. ADFT was lower in acute AF than in chronic AF. ADFT was highly reproducible during repeated defibrillation. Sotalol reduced ADFT in acute AF and to a lesser extent in chronic AF, and increased the defibrillation success rate. Ventricular pacing will often be required because of prolonged pause after successful defibrillation. PMID- 9358488 TI - Effect of ventricular pacing on coronary blood flow in patients with normal coronary arteries. AB - Although ventricular pacing is thought to produce impairment of left ventricular function by altering the sequence of ventricular activation and AV dyssynchrony, little is known about the effect of ventricular pacing on coronary blood flow. We measured coronary blood flow and coronary flow reserve in the left anterior descending coronary artery during sinus rhythm, and during both atrial and ventricular pacing at a rate of 100 ppm in 14 patients with normal coronary arteries. The double product increased significantly during both types of pacing. Coronary arterial diameter during ventricular pacing significantly increased compared to that during both sinus rhythm and atrial pacing. Coronary flow velocity during ventricular pacing was significantly lower compared to that during both sinus rhythm and atrial pacing. Coronary blood flow increased significantly during atrial pacing (30.7% +/- 12.1%; P < 0.001), but not significantly during ventricular pacing (23.6% +/- 47.0%; P = ns). While coronary flow reserve during both atrial (3.9 +/- 1.3) and ventricular pacing (3.8 +/- 0.9) was lower compared to its value during sinus rhythm (4.5 +/- 1.5), the difference was not significant. There was a significant positive correlation between the coronary flow reserve during sinus rhythm and the increase of coronary blood flow during ventricular pacing (R2 = 0.78; P < 0.001). We concluded that an increase in coronary blood flow during ventricular pacing is not a common finding regardless of the increase in metabolic demand. The increase of coronary blood flow during ventricular pacing was less in patients with a reduced coronary flow reserve. These findings suggest that preservation of AV synchrony and the presence of a normal sequence of ventricular activation may play an important role in preserving coronary blood flow in this subset of patients. PMID- 9358487 TI - Optimizing the AV delay in DDD pacemaker patients with high degree AV block: mitral valve Doppler versus impedance cardiography. AB - In DDD-pacemaker patients with high degree AV block, Doppler echocardiography of transmitral blood flow can be used to find the individually optimal AV delay (AVO) for left heart AV synchronization. This study tried to validate a Doppler method (ECHO) recently proposed to optimize left ventricular filling by comparing it to stroke volume data derived from impedance cardiography (ICG). It should be further elucidated if optimizing the AV delay (AVD) by means of this method is superior to fixed AVD settings and which differential AVD (pace-sense-offset) should be programmed for atrially triggered (ATP) and AV sequential (AVP) pacing, respectively. AVO as measured in 53 patients showed a linear correlation between ECHO and ICG for both ATP (r = 0.66, P < 0.00001) and AVP (r = 0.53; P < 0.005). The mean deviation in AVO between ECHO and ICG was +/- 26 ms (ATP) and +/- 30 ms (AVP), respectively, with a tendency to longer AVDs with the Doppler method. ECHO limitations could mainly be attributed to: (1) restrictions of AVD programming options (which may be compensated for by slight modification of the proposal); and (2) to pathophysiological mechanisms that alter mitral valve dynamics. Optimization of the AVD by Doppler produced a stroke volume that was significantly higher (19%) than with a fixed AVD (150 ms in ATP; 200 ms in AVP). There was a wide scatter in pace-sense-offsets between-7 and 134 ms, which was reflected by both methods. It is concluded that AVO determinations by ECHO are valid provided that methodological pitfalls and limitations caused by the disease are recognized. Tailoring AVD with respect to diastolic filling improves systolic function and is superior to nominal AVD settings. Fixed differential AVDs as offered by some manufacturers are far from being physiological. Thus modern pulse generators should offer free programmability over a wide range of AV delays. PMID- 9358489 TI - Analysis of electrophysiological activity in Koch's triangle relevant to ablation of the slow AV nodal pathway. AB - Atrioventricular junctional reentrant tachycardia (AVJRT) is the most common form of paroxysmal regular supraventricular tachycardia. In patients with disabling, drug refractory AVJRT, catheter ablation has evolved rapidly from a last-resort treatment in the form of interruption of atrioventricular (AV) conduction to selective modification of AV nodal function as an ideal treatment. This article will focus on the frequently unappreciated electrophysiological activities recordable in man in Koch's triangle during ablation of the so-called slow pathway. PMID- 9358492 TI - Use of an atrial loop to extend the duration of endocardial pacing in a neonate. AB - Use of an atrial loop has been proposed as a means of extending the longevity of endocardial pacing systems in small children who require ventricular pacing. A few reports have demonstrated the effectiveness of this method at reducing the number of interventions in infants and small children, but there is little reported experience in neonates. Permanent endocardial ventricular demand pacing was performed in a 3.4-kg neonate. The generator was placed in a prepectoral pocket and a redundant loop of lead was left in the atrium to cater for further growth. At 32 months he weighs 13.6 kg and the loop has uncoiled leaving additional lead slack for further growth. PMID- 9358490 TI - Syncope in the presence of newly developed bundle branch block: bradycardia or tachycardia related. PMID- 9358491 TI - A left free-wall, decrementally conducting, atrioventricular (Mahaim) fiber: diagnosis at electrophysiological study and radiofrequency catheter ablation guided by direct recording of a Mahaim potential. AB - A 64-year-old female with Wolff-Parkinson-White syndrome and an ECG demonstrating a right posterolateral accessory pathway was referred for electrophysiological study. During electrophysiological testing two AV pathways were identified: a right posterolateral pathway that displayed conventional electrophysiological properties: and a left free-wall pathway that conducted only anterogradely and demonstrated decremental properties. Two separate wide complex tachycardias were induced that utilized the left free-wall pathway anterogradely and either the AV node or the right posterolateral accessory pathway retrogradely. A discrete electrical potential on the free wall of the mitral annulus was identified during tachycardia and was utilized to facilitate mapping and ablation. PMID- 9358493 TI - False-positive behavior with the dP/dt sensing pacemaker: a rare complication of a physiological sensor. AB - As with "nonphysiological" devices, sensors that directly measure physiological variables have the potential to measure unexpected signals and for the physiological parameter being measured to respond in an unexpected manner. We present the case of a dP/dt sensing pacing system that functioned normally for 2 months and then developed upper rate behavior due to the sensing of a high frequency artifact on the pressure recording. Our case and others cited reinforce the need for future physiological rate responsive pacemakers to incorporate a second sensor to provide for backup rate response in cases of inappropriate rate response. PMID- 9358494 TI - Supernormality with cycle length-dependent intra-His alternans: a new cause of group beating. AB - This article describes a patient with baseline infranodal conduction disease. Following the administration of intravenous procainamide, the patient developed group beating. Careful electrophysiological evaluation revealed that the group beating was due to alternating intra-His conduction times. Systematic analysis of paced and spontaneous events provides strong evidence that the alternans results from supernormality within the His bundle. PMID- 9358495 TI - Transient data stream dissociation in computerized data acquisition system masquerading as a sensing abnormality. AB - During testing of a CPI model 1715 ICD, an apparent sensing abnormality was noted following shock delivery for VF. Close inspection of the recording prior to the defibrillation attempt revealed that the surface leads spontaneously lost 848 ms of data while the event marker was unaffected. Computer simulations revealed that an inadequate buffer size for the amplified (surface ECG) data was the likely source of data loss. It is important to recognize that a discordance between surface leads and event marker may represent an abnormality in the data acquisition system and simulate an ICD or lead malfunction. PMID- 9358496 TI - Radiofrequency ablation of a posteroseptal accessory pathway via the middle cardiac vein in a six-year-old child. AB - Although radiofrequency ablation is highly successful in patients with the Wolff Parkinson-White syndrome, certain pathways remain refractory to ablation. In particular, subepicardial pathways often fail ablation via an endocardial approach. In adult patients, left-sided subepicardial pathways have been treated successfully using energy delivery within the coronary sinus. To document the safety and efficacy of this approach in children, we present the case of a 6-year old boy who underwent radiofrequency ablation of a posteroseptal pathway via energy delivery within the middle cardiac vein. Follow-up study showed no evidence of recurrence or gross coronary vascular injury. PMID- 9358497 TI - Banning unipolar leads. PMID- 9358498 TI - Clinical perspective on risk stratification. AB - Surveys show that subjective risk assessment or risk stratification is often widely inaccurate. Objective data from large observational studies or from clinical trials identifies a persons absolute risk of an event in a given time in order to assess the risk/benefit ratio of a given treatment. In general, the higher the risk the better the risk/benefit ratio. Relative risk reduction by a given treatment is often similar across subgroups divided by sex, age, blood pressure etc.; however if the absolute risk is low it may not e worth taking a treatment with serious side effects (e.g., cerebral haemorrhage). Risk stratification is also important in assessing the cost effectiveness of treatment (e.g,. cholesterol reduction by statins for different groups of the population). Inappropriate surrogate end points should be avoided in cost benefit analysis (e.g., suppression of ventricular ectopic beats by antiarrhythmic drugs). A Bayesian approach should be adopted. PMID- 9358499 TI - Risk assessment and risk stratification in sudden cardiac death: a biostatistician's view. AB - Determining individual probabilities of developing lethal arrhythmia over time (risk assessment) and grouping individuals by that probability (risk stratification) are similar to, yet differ in purpose from, screening, diagnosis, risk factor identification, and prognostic staging. Methods of handling bias, use of multiple predictors, and evaluation of results provide challenges. A key purpose of risk assessment and stratification is examined. The role of operational definitions of predictors and events and of methods that account for multiple predictors and known confounding factors is analyzed. Constructed examples illustrate potential pitfalls in assessment and how multivariate techniques can deal with multiple predictors. A trial design to evaluate risk stratification for the identified purpose is elaborated and potential results are interpreted. Bias from predictors regressing to the mean can be minimized either by averaging a number of measurements or by equalizing the bias in comparison groups. An analysis of two predictors and two risk strata illustrates how the discrimination of combined predictors may be greater than the sum of the individual variables' discrimination. Risk stratification can be evaluated in trials that randomize competing interventions within different risk strata. Results of such trials indicate whether the risk strata adequately distinguish individuals by their responsiveness to particular intervention. Potential pitfalls, not easily recognized in risk stratification, can be avoided in the methods and in studies for evaluating those methods. Multivariate techniques maximize the discrimination of multiple predictors, but may increase complexity. Randomized trials of treatment provide evidence for utility of risk stratification. PMID- 9358501 TI - Classification of sudden and arrhythmic death. AB - Since all death is (eventually) sudden and associated with cardiac arrhythmias, the concept of sudden death is only meaningful if it is unexpected, while arrhythmic death is only meaningful if life could have continued had the arrhythmia been prevented or treated. Current classifications of death as being arrhythmic or sudden are all biased by the difficulty of having to decide on the degree of unexpectedness or the likelihood that life could continue without the arrhythmia. The uncertainties are enlarged by the fact that critical data (such as knowledge of arrhythmias at the time of death or autopsy) are available in only a few percent of cases. A main problem in using classifications is the lack of validation data. This situation has, with the MADIT trial, changed in the case of the Thaler and Hinkle classification of arrhythmic death. The MADIT trial demonstrated that arrhythmic death was nearly abolished by the implantable defibrillator, indicating that arrhythmic death by this classification is meaningful, at least in the population studied. For future investigations, a call is made for committees to present data in a way that allows the reader to examine the quality of the data used for evaluation. PMID- 9358500 TI - Analysis of clinical follow-up databases: risk stratification studies and prospective trial design. AB - Design of new prospective studies should utilize detailed retrospective evaluations of clinical data. For this purpose, clinical data are needed containing both prospectively recorded values of risk factors and follow-up events. A concept of a new trial can then be modeled within the existing data set. The development of such a model consists of the following steps: (a) the distribution of values of risk factors has to be investigated in the whole recorded population and the statistical association of the risk factors with follow-up events has to be established; (b) the stratification characteristics (sensitivity, specificity, and predictive accuracies) have to be evaluated for individual risk factors and for their multivariate combinations; (c) the stratification characteristics have to be converted into estimates of mortality reduction expected within the high risk group and used for the optimum trial design in terms of screened and randomized patient numbers. In this review, the strategy of designing a new trial is demonstrated using data of 644 survivors of acute myocardial infarction with available 3-year follow-up during which 74 patients died. A model of a new trial is considered involving reduced left ventricular ejection fraction, increased 24-hour mean heart rate, and depressed 24-hour heart rate variability as risk stratifiers. PMID- 9358502 TI - Use of left ventricular ejection fraction or wall-motion score index in predicting arrhythmic death in patients following an acute myocardial infarction. The TRACE Study Group. AB - All-cause mortality and morbidity following an acute myocardial infarction (AMI) are correlated to LV systolic dysfunction. The correlation is closest with mortality and morbidity associated with congestive heart failure (CHF). Prediction of arrhythmic death in patients with AMI relies on the correlation between arrhythmic death and "sudden unexpected death" defined as death within 1 hour of onset of new symptoms. Assessment of late potentials, heart rate variability (HRV), T wave alternans, arrhythmias seen on Holter monitoring or during exercise testing, electrophysiological testing, and baroreceptor assessment have all proven to be useful in the prediction of sudden death even when LV systolic function is known. In selected populations HRV is superior to LV systolic function assessment in predicting sudden death and/or arrhythmic events, and may even predict all-cause mortality with the same precision. Comparisons of other methods with LV function assessment should be interpreted with care because most methods have been evaluated in subgroups of infarct patients with a low risk of death. Results from a large series of high risk patients with AMI (the TRAndolapril Cardiac Evaluation study) have shown that even in patients with severe depressed LV systolic function around one-third of the patients will die suddenly. The current situation is that LV function appears to be the best method of predicting death whereas other methods appear very promising for detecting arrhythmic death in more selected populations. The optimal method for selecting patients at high risk of arrhythmic death has not yet been developed, but a combination of LV function and another method, i.e., HRV, appears promising. This may ensure that the enrolled patients have an increased risk of death and that this risk will be due to arrhythmic events. Patients with LVEF of 10% or less can be excluded as they will most likely not die suddenly. PMID- 9358505 TI - Signal-averaged P wave in patients with paroxysmal atrial fibrillation. AB - The theoretical and experimental rational of atrial signal-averaged ECG in patients with AF is delay in the intra-atrial and interatrial conduction. Similar to the ventricular signal-averaged ECG, the atrial signal-averaged ECG is an averaging of a high number of consecutive P waves that match the template created earlier P wave triggering is preferred over QRS triggering because of more accurate aligning. However, the small amplitude of the atrial ECG and its gradual increase from the isoelectric line may create difficulties in defining the start point if P wave triggering is used. Studies using P wave triggering and those using QRS triggering demonstrate a prolonged P wave duration in patients with paroxysmal AF. The negative predictive value of this test is relatively high at 60%-80%. The positive predictive value of atrial signal-averaged ECGs in predicting the risk of AF is considerably lower than the negative predictive value. All the data accumulated prospectively on the predictive value of P wave signal-averaging was determined only in patients undergoing coronary bypass surgery or following MI; its value in other patients with paroxysmal AF is still not determined. The clinical role of frequency-domain analysis (alone or added to time-domain analysis) remains undefined. Because of this limited knowledge on the predictive value of P wave signal-averaging, it is still not clinical medicine, and further research is needed before atrial signal-averaged ECG will be part of clinical testing. PMID- 9358504 TI - Prediction of arrhythmia risk based on signal-averaged ECG in postinfarction patients. AB - In patients surviving acute MI, identification of those at high risk for life threatening ventricular tachyarrhythmias and/or sudden death is of great importance. Numerous strategies based on indices such as the degree of left ventricular dysfunction, complex ventricular arrhythmias, or parameters of autonomic dysfunction have not yet led to an effective identification of the individual patient at risk. During the past decade, many investigators have recorded low amplitude, high frequency components in the terminal QRS complex (so called late potentials) from patients prone to sustained ventricular tachycardia. The SAECG has been used to predict life-threatening tachyarrhythmias in patients after acute MI and to screen for inducible ventricular tachycardia in patients with unexplained syncope or sustained ventricular tachycardia. This review article describes the most frequently applied methodology and clinical applications of the SAECG in post-MI patients and discusses the usefulness of noninvasive recordings in various other clinical settings. PMID- 9358506 TI - Present and future role of ambulatory Holter monitoring for arrhythmia risk stratification. AB - Risk stratification for arrhythmogenic events and sudden death in patients with organic heart disease, particularly those with coronary heart disease and a history of MI, continues to be one of the major tasks of clinical cardiologists, although advanced management strategies including thrombolysis, acute PTCA and surgical intervention dramatically reduced the percentage of sudden deaths following acute MIs, Noninvasive studies like resting and exercise ECG, echocardiography, signal averaging, 24-hour ECG, and radionuclide studies, as well as invasive techniques such as electrophysiologically programmed electrostimulation and coronary angiography, are being used routinely. Ambulatory Holter monitoring is an established noninvasive technique for risk stratification. There is evidence showing that its predictive potential for arrhythmogenic risks is enhanced, if more than one parameter is analyzed. Absence of ST segment changes and a normal HRV are the parameters signaling out low-risk patients. The use of additional parameters which escape electrocardiographic recording, like ventricular function and myocardial ischemia, improve the accuracy of predicting arrhythmogenic events. The most predictive combination of risk parameters is, however, still poorly understood. Future research should define normal ranges of parameters recordable by H-ECG, solve technical problems of recording data and analyzing them. In addition, the accuracy of measuring QT duration and documenting late potentials should be improved by more sophisticated methods. But it is unrealistic to expect that the QT interval will become amenable to automatic analysis in all patients. A fully automatic QT analysis without visually checking the measuring points at the tip and the end of the T wave for their consistency is hardly conceivable. The documentation of late potentials, in turn, is limited by artefacts caused by muscle contraction during physical activity. Clinical aspects, e.g., the predictability of arrhythmogenic events in patients with cardiomyopathies and valvular disease should be addressed. This will require studies combining the predictive potentials of rhythmologic and hemodynamic data. PMID- 9358507 TI - Heart rate variability used as an arrhythmia risk stratifier after myocardial infarction. AB - Heart rate variability (HRV) is considered to represent a noninvasive tool to assess cardiac autonomic tone at the level of the sinus node. It has been shown to have predictive power for risk assessment in patients surviving acute myocardial infarction. For this purpose, HRV should be assessed from 24-hour Holter recordings obtained 7-10 days following the infarction. Although there is some recovery of HRV during the first 3 months after infarction, HRV remains reduced in postinfarction patients compared to values obtained in healthy individuals. Compared to assessment of left ventricular function as a risk marker, HRV is superior with respect to prediction of arrhythmic events and sudden death whereas both parameters yield comparative power for prediction of total cardiac mortality. Since the predictive power of HRV analysis alone is relatively low, the combined use of HRV measurements together with traditional risk markers (such as ventricular ectopic beats, signal-averaged ECG, or left ventricular function) results in improved risk prediction with positive predictive accuracy in the range of 30%-50%. PMID- 9358508 TI - Baroreflex sensitivity as a cardiac and arrhythmia mortality risk stratifier. AB - The last two decades have provided clear evidence for the tight and casual relation existing between arrhythmic mortality and the autonomic nervous system, particularly with imbalances characterized by decreases in vagal and/or increases in sympathetic activity. A series of compelling experimental results has represented the driving force for the clinical evaluation of the potential prognostic value of baroreflex sensitivity (BRS), a measure that can provide information on the capability to augment vagal activity. This article reviews the methodology more commonly used to quantify the clinical evaluation of this parameter, and then focuses on the key clinical studies highlighting those performed in postmyocardial infarction patients. Among them the most informative is ATRAMI, a multicenter prospective study involving almost 1300 patients. The main conclusion is that both heart rate variability and BRS are strong and independent risk factors for post-infarction mortality, thus demonstrating the clinical usefulness of autonomic markers. PMID- 9358503 TI - Arrhythmia risk: electrophysiological studies and monophasic action potentials. AB - Shortly after in the introduction of programmed electrical stimulation (PES) of the heart to study and localize cardiac arrhythmias in the intact human heart, the technique was used for risk stratification of the arrhythmia patient. Two decades later we have to conclude that especially in ventricular arrhythmias the technique of PES did not live up to our expectations and the left ventricular function is a better long-term predictor than the induction of ventricular arrhythmias or the ability to find an antiarrhythmic drug able to prevent the initiation of the classically documented ventricular arrhythmia. Another sobering finding came from the analysis of the characteristics of the patient dying suddenly out-of-hospital, which showed that most of those patients could not be classified before the event as being at high risk using noninvasive or invasive testing, not even in those with a previous cardiac history. Monomorphic action potential (MAP) recordings have been of importance in our understanding of torsade de pointe arrhythmias in congenital and acquired QT prolongation. A major problem in case of a less generalized electrophysiological abnormality is the identification of the appropriate place where to put the MAP-electrode. PMID- 9358510 TI - QT interval dispersion and its clinical utility. AB - QT dispersion as a measure of interlead variations of QT interval duration in the surface 12-lead ECG is believed to reflect regional differences in repolarization heterogeneity and thus, may provide an indirect marker of arrhythmogenicity. Methodology for determining QT dispersion and reproducibility of this parameter vary significantly between studies and, together with some other unresolved problems with QT dispersion assessment, often lead to contradictory suggestions about potential clinical utility of this parameter. The results of our own study in 213 survivors of myocardial infarction, together with a comprehensive review of the literature, suggest that most of these inconsistencies reflect incomplete understanding of electrocardiographic correlates of both normal and abnormal ventricular repolarization. The application of more objective techniques, such as spectral analysis or combined assessment of different parameters (e.g., area beneath the T wave and its symmetricity) may add a new dimension to the noninvasive assessment of ventricular repolarization. PMID- 9358509 TI - Changes in repolarization dynamicity and the assessment of the arrhythmic risk. AB - At the present time, the assessment of the arrhythmic risk from surface ECG recordings is built on time-domain and frequent-domain analysis of high resolution ECG acquisition together with interlead variability of QT interval duration (QT dispersion). The corresponding raw ECG tracings are obtained in resting conditions. However, the dynamic aspects of the ECG signal is a rapidly evolving matter of interest. In addition to the beat-to-beat oscillations of the ventricular repolarization amplitude (QT alternans), there is growing evidence that the patterns of QT interval shortening with increasing heart rate are linked to susceptibility to ventricular arrhythmias. In this report, we will mainly address the association between QT dynamicity and the risk of developing torsades de pointes. PMID- 9358511 TI - Repolarization alternans: techniques, mechanisms, and cardiac vulnerability. AB - Sudden cardiac death continues to be the leading cause of mortality in developed countries. Electrical alternans of the ST segment and the T wave on the surface ECG as a noninvasive marker of patients at risk is a phenomenon that was initially observed early in this century and was seen then to be associated with cardiac rhythm disturbances. Substantial evidence indicates that T wave alternans (TWA) is related to myocardial ischemic as a harbinger of malignant ventricular arrhythmias because it reflects dispersion and heterogeneity of repolarization. Recent data have demonstrated a significant correlation between TWA and vulnerability to ventricular arrhythmias in individuals with or without organic heart disease, it also predicts the results of electrophysiological testing and arrhythmia-free survival in patients with a variety of cardiac diseases. This article reviews the historical background of TWA and discusses the early experimental and recent clinical evidence implying an integral link between TWA and ischemia-induced cardiac vulnerability. PMID- 9358512 TI - Holter, loop recorder, and event counter capabilities of implanted devices. AB - The current generation of cardiac pacemakers and implantable cardioverter defibrillators almost all have some capabilities to store data regarding device activity and patient events for future retrieval. This information may provide valuable information regarding device function and whether this is proving valuable in patient management. Examples include "pace-sense" counters, which can reveal under sensing of patient events, and serial lead impedance measurements, which are able to demonstrate trends not seen on isolated measurements. Holter capabilities become vital in more advanced devices for documenting the utility of, and fine tuning the programming of features such as antitachycardia pacing, rate-responsiveness, and mode-switching. Finally, the ability to store patient events as marker channels and even intracardiac electrograms adds a diagnostic capability not available through external monitoring. This role has now been advanced by the development of a purely diagnostic implantable loop recorder. PMID- 9358513 TI - Factors predisposing to the development of atrial fibrillation. AB - Atrial fibrillation (AF) is in most patients (approximately 70%) associated with organic heart disease including valvular heart disease, coronary artery disease, hypertension, hypertrophic cardiomyopathy, dilated cardiomyopathy, and congenital heart disease, mostly atrial septal defect in adults. In many chronic conditions, determining whether AF is the result or is unrelated to the underlying heart disease, remains unclear. The list of possible etiologies also include cardiac amyloidosis, hemochromatosis and endomyocardial fibrosis. Other heart diseases, such as mitral valve prolapse (without mitral regurgitation), calcifications of the mitral annulus, atrial myxoma, pheochomocytoma, and idiopathic dilated right atrium may present with AF. Atrial fibrillation may occur in the absence of detectable organic heart disease, the so-called "lone AF", in about 30% of cases. The term "idiopathic AF" implies the absence of any detectable etiology including hyperthyroidism, chronic obstructive lung disease, overt sinus node dysfunction, and overt or concealed preexcitation (Wolff-Parkinson-White syndrome), only to mention a few of other uncommon causes of AF. The autonomous nervous system may contribute to the occurrence of AF in some patients. AF occurs commonly. In patients with valvular heart disease, AF is common, particularly when the mitral valve is involved. The occurrence of AF is unrelated to the severity of mitral stenosis or mitral regurgitation but is more common in patients with enlarged left atrium and congestive heart failure. In patients with coronary artery disease, AF occurs predominantly in older patients, males, and patients with left ventricular dysfunction, Important predictive factors of AF include hypertension, left ventricular hypertrophy and diabetes. The risk of the development of AF, in an individual patient, is often difficult to assess. Increasing age, presence of valvular heart disease, and congestive heart failure increase the risk of atrial fibrillation. PMID- 9358514 TI - Cardioversion of atrial fibrillation and subsequent maintenance of sinus rhythm. AB - This article gives an overview of electrical cardioversion of AF and includes the discussion of newer strategies. DC external cardioversion is highly effective and carries a low risk of complications. Other approaches, like transesophageal cardioversion and high energy internal cardioversion, may improve the acute success rate but do not enhance long-term maintenance of sinus rhythm compared to external cardioversion. An atrial defibrillator may have important advantages which relate to the fact that the duration and possibly also the number of AF episodes become reduced. Supposedly, shortening the attacks of AF may exert an antiarrhythmic effect by limiting electrical, anatomical, and neurohumoral remodeling. So far, low energy biatrial defibrillation using an atrial defibrillator seems to be effective and safe (i.e., does not induce ventricular arrhythmias). However, discomfort limits its tolerability in clinical practice. Future improvement of leads and light sedation that is easy to administer may overcome this problem. In the second part of this overview, the probability of AF recurrence using a serial cardioversion approach is discussed. In middle-aged patients with a fair exercise tolerance and an arrhythmia duration < than 36 months this approach may be worthwhile. Young patients (age < 57 years) with an arrhythmia duration < 3 months and without hypertension may be cured from the arrhythmia with a single shock and without the institution of antiarrhythmic drugs. However, the serial electrical cardioversion approach is unlikely to succeed in elderly patients with a duration of AF exceeding 36 months and a poor exercise tolerance (NYHA Functional Class III or IV). PMID- 9358515 TI - Risk of complications of atrial fibrillation. AB - Atrial fibrillation is associated with three major risk of complications: thromboembolism, hemodynamic compromise, and arrhythmogenesis. In patients with chronic atrial fibrillation the incidence of embolization is about 5% per year. The risk of embolism and in particular of stroke can be reduced by warfarin anticoagulation. Aspirin is generally less effective than warfarin, although it is probably more effective than placebo. The hemodynamic complications which may occur during atrial fibrillation are mainly due to the loss of effective atrial contraction, the irregular ventricular rhythm, and the possible excessively rapid ventricular rate. Sudden death is a recognized manifestation of Wolff-Parkinson White syndrome and is considered to be precipitated by atrial fibrillation in the majority of patients. Torsades de pointes is perhaps the most widely recognized proarrhythmia associated with treatment of atrial fibrillation, especially with 1A antiarrhythmic drugs and sotalol. The chronic treatment with type 1C drugs in 3.5%-5% of patients may induce atrial flutter with 1:1 conduction with significant hemodynamic compromise. PMID- 9358516 TI - Ventricular arrhythmias in apparently healthy subjects. AB - While it is assumed that the normal heart does not predispose to serious arrhythmias, several conditions are now being recognized as being associated with short-lasting ventricular arrhythmias. It also becomes clear that idiopathic VT (or repetitive monomorphic VT) sometimes exists on the background of a compromised heart. Whether this dysfunction is due to the arrhythmia or vice versa is not evident. Finally, VF occurs in patients who, at a first glance, have no apparent heart disease, and it is then called idiopathic VF. These complex electrical abnormalities probably reflect disorders, which often are genetically determined. Recognition of these syndromes, often characterized by abnormal repolarization or a disturbed autonomic function is possible if appropriate techniques are used. PMID- 9358517 TI - Risk of ventricular arrhythmias in survivors of myocardial infarction. AB - The most recent studies have made it clear that the prognosis of asymptomatic post-MI patients has significantly improved in the last two decades. Holter monitoring as well as a low LVEF still is an important method for the risk stratification in the thrombolytic era of patients with post-MI. Patients with normal noninvasive tests do have a good prognosis. The electrophysiological stimulation seems to be the clinically most valuable single method to predict arrhythmic events. However, as an invasive procedure it is not suitable as a screening test for a large cohort. The stepwise risk stratification technique using first noninvasive followed by invasive procedures seem to be most suitable and effective for identifying asymptomatic infarct survivors which incidence of arrhythmic events is as high as the recurrence rate of patients who had been resuscitated from ventricular fibrillation. Consequently, prophylactic implantation of a defibrillator in asymptomatic MI patients, whose positive predictive value is around 30% becomes more and more interesting. PMID- 9358518 TI - Arrhythmias in hypertrophic cardiomyopathy. AB - Supraventricular and ventricular arrhythmias, particularly nonsustained ventricular tachycardia, and ventricular premature beats are a common finding in patients with hypertrophic cardiomyopathy. Several investigations have demonstrated that nonsustained ventricular tachycardia on Holter monitoring is associated with an increased risk of sudden cardiac death. It has been a long lasting controversial discussion whether suppression of these arrhythmias with drugs, such as amiodarone is capable to reduce the incidence of sudden cardiac death. While recent studies have indicated that nonsustained ventricular tachycardia in asymptomatic patients without additional risk factors, such as a positive family history of sudden cardiac death or syncope should not be treated prophylactically with amiodarone. Symptomatic patients with sustained ventricular tachycardias and/or syncope related to ventricular arrhythmias should undergo ICD implantation. The implantation of an ICD in asymptomatic patients should be limited to those who have several risk factors for sudden cardiac death. It is questionable whether other risk stratifiers, such as programmed electrical stimulation may be helpful to identify asymptomatic patients who are at risk to die suddenly. Moreover, whether the demonstration of electrocardiogram fractionation during electrophysiological study is superior to the induction of sustained ventricular arrhythmias for risk stratification, needs further investigation. PMID- 9358519 TI - Ventricular arrhythmias in dilated cardiomyopathy. AB - Although prognosis of dilated cardiomyopathy (DCM) has improved due to advances in diagnosis and therapy, still too many sudden cardiac deaths occur in DCM. Spontaneous ventricular ectopy is a very common finding in patients with DCM, but the prognostic significance of Holter monitoring remains controversial. Other noninvasive methods, e.g., late potentials and QT dispersion, have not yet contributed to the evaluation of prognosis for arrhythmogenic events in DCM. Programmed ventricular stimulation has been repeatedly used to stratify long-term prognosis, yet satisfactory data are still missing as many deaths occur in patients without inducible arrhythmias. Several prognostic studies are still in progress, and preliminary data for the use of ICDs already appear to be promising. In patients with poor left ventricular function and ICDs in situ, prognosis is determined by progression of heart failure. Heart transplantation may be the ultimate therapeutic instrument for end-stage heart failure patients. For patients with advanced DCM and increased risk for malignant arrhythmias who are unsuitable for orthotopic heart transplantation, the combined therapy with an ICD and dynamic cardiomyoplasty may be an alternative treatment. PMID- 9358520 TI - Management of patients with life-threatening ventricular tachyarrhythmias in the defibrillator era: the need to differentiate. AB - Patients with a history of sustained ventricular tachyarrhythmias form an extremely inhomogeneous group with respect to presenting arrhythmia, underlying cardiac disease, and therefore, risk of dying suddenly. For subgroups such as ventricular tachycardia in the absence of underlying cardiac disease, radiofrequency catheter ablation offers cure. In others, implantation of a cardioverter defibrillator already appears to have gained the therapy of first choice, leaving only a secondary role to antiarrhythmic drugs. It must be emphasized however, that these new therapeutic strategies have their pros and cons like the older, seemingly out-fashioned approaches of noninvasively or invasively guided antiarrhythmic drug therapy or empiric amiodarone treatment. Until the advent of controlled randomized trials comparing the implantable cardioverter defibrillator (ICD) with the best other, usually medical form of treatment, physicians must continue to base their individual therapeutic decisions on circumstantial published and personal experience. In doing so, the recent achievements of catheter ablation and defibrillator implantation have definitely improved patient care, but have not made antiarrhythmic drugs jobless. With all the alternatives at hand, it remains a challenging task to weigh the benefits and risks of the various approaches against each other in an attempt to tailor the antiarrhythmic intervention to the very individual need of the patient. PMID- 9358521 TI - Early initiation of chronic dialysis: role of incremental dialysis. PMID- 9358522 TI - The problem of mathematical coupling: how can statistical artifact and biological causation be separated when relating protein intake to clearance in 'predialysis' and dialysis patients? PMID- 9358523 TI - Will automated peritoneal dialysis be the answer? PMID- 9358524 TI - Uremic pruritus in CAPD patients. PMID- 9358525 TI - Adequacy of dialysis in automated peritoneal dialysis: a clinical experience. AB - OBJECTIVES: To compare the peritoneal clearances of urea and creatinine in continuous ambulatory peritoneal dialysis (CAPD) with three types of automated peritoneal dialysis (APD): continuous cycling peritoneal dialysis (CCPD), 50% tidal peritoneal dialysis (TPD), and 25% TPD and to assess the usefulness of the peritoneal equilibration test (PET) in predicting peritoneal clearances in overnight APD. PATIENTS: Eleven uremic patients (mean age 44.5 +/- 15.45 years with a mean time on dialysis of 42.63 +/- 25.62 months) were included in the study. MEASUREMENTS: PET for urea and creatinine following Twardowski's method. Peritoneal clearances for urea and creatinine CAPD: samples of blood and dialysate within 24 hours. APD: blood mean levels of urea and creatinine before and after nighttime dialysis. Dialysate: urea and creatinine in nocturnal and daytime dialysate. RESULTS: Peritoneal clearance of creatinine was 38.14 +/- 9.99 L/week/1.73 m2 in CAPD, 44.28 +/- 12.4 L/week/1.73 m2 in CCPD, 50.07 +/- 17.86 L/week/1.73 m2 in 50% TPD (p < 0.05) and 40.18 +/- 6.65 L/week/1.73 m2 in 25% TPD. Peritoneal clearance of urea improved significantly in the three modalities of APD: 51.91 +/- 12.58 L/week/1.73 m2 in CAPD; 66.7 +/- 9.9 L/week/1.73 m2 in CCPD (p < 0.05); 76.3 +/- 14.5 L/week/1.73 m2 in 50% TPD (p < 0.001) and 64.4 +/- 11.4 L/week/1.73 m2 in 25% TPD (p < 0.05). The dialysate/ plasma (D/P) ratio of creatinine at 30, 60, 120, 180, and 240 minutes showed significant correlation with nighttime APD clearance. Nevertheless, only the D/P ratio of urea at 30, 60, and 120 minutes correlated with overnight APD clearance. CONCLUSIONS: A remarkable improvement was observed with APD regarding the clearance of urea mainly when 50% tidal peritoneal dialysis was used, whereas it was less noticeable in the clearance of creatinine. The PET is a helpful tool in predicting overnight peritoneal clearances of creatinine but it is less useful in predicting urea clearance. PMID- 9358526 TI - Effect of dialysis modality and membrane transport characteristics on dialysate protein losses of patients on peritoneal dialysis. AB - OBJECTIVE: To determine if peritoneal dialysis modality has an impact on protein losses in dialysate. DESIGN: Retrospective, cross-sectional study. PATIENTS: 190 patients who had selected peritoneal dialysis were classified into one of four transport categories (high, high-average, low-average, or low) based on standard peritoneal equilibration test results. Patients were then assigned to continuous ambulatory peritoneal dialysis (CAPD) or nightly intermittent peritoneal dialysis (NIPD) based on membrane transport characteristics and individual preferences. RESULTS: Patients with similar membrane transport characteristics had essentially no differences in dialysate protein and albumin losses whether treated with CAPD or NIPD. CONCLUSIONS: Although high transporters may be better managed with short dwell therapies such as nocturnal intermittent peritoneal dialysis or daily ambulatory peritoneal dialysis, consistent marked decreases in protein losses cannot be cited as a benefit of NIPD over CAPD. PMID- 9358527 TI - Absence of modulating effects of cytokines on antioxidant enzymes in peritoneal mesothelial cells. AB - OBJECTIVE: To investigate the modulation of superoxide dismutase, glutathione peroxidase, and catalase by cytokines and endotoxin in human peritoneal mesothelial cells. DESIGN: Cultured human peritoneal mesothelial cells were treated with various concentrations of interleukin-1 alpha, tumor necrosis factor alpha (TNF-alpha), interleukin-6, interleukin-8, transforming growth factor-beta (TGF beta), and lipopolysaccharide. Cell morphology was observed and the activities of superoxide dismutase, catalase, and glutathione peroxidase were assayed. The antioxidant enzyme activities of human peritoneal mesothelial cells were also compared with those of human liver and kidney tissues. RESULTS: Interleukin-1 alpha, TNF alpha, TGF beta, and lipopolysacharide caused dose dependent cytotoxicities in mesothelial cells. The activities of these three antioxidant enzymes did not change after treatment with cytokines and endotoxin. The total superoxide dismutase activity of confluent human peritoneal mesothelial cells was found to be greater than that of human liver and kidney tissues and was composed mostly of manganese superoxide dismutase activity. Furthermore, glutathione peroxidase and catalase activities of human peritoneal mesothelial cells were lower than those of human liver and kidney tissues. CONCLUSION: In human peritoneal mesothelial cells, lack of induction of antioxidant enzymes by inflammatory cytokines, as well as high superoxide dismutase activity accompanied by insufficient glutathione peroxidase and catalase activities may both contribute to the susceptibility of these cells to oxidative damage. Therefore, appropriate management to decrease oxidative injury to the peritoneum should be taken into consideration when treating long-term continuous ambulatory peritoneal dialysis patients. PMID- 9358528 TI - Increase of the bioavailability of intraperitoneal erythropoietin in children on peritoneal dialysis by administration in small dialysis bags. AB - OBJECTIVE: To establish the effectivity of administration of erythropoietin intraperitoneally in a small amount of fluid in children with renal anemia on continuous ambulatory peritoneal dialysis (CAPD). DESIGN: Prospective study in which children with renal anemia on CAPD were treated with erythropoietin intraperitoneally, administered in a specially designed bag containing 50 mL NaCl 0.9%. SETTING: University hospital. PATIENTS: The patient population consisted of 9 children treated with CAPD and 1 treated with nightly intermittent peritoneal dialysis. The median age was 7.8 years (range 4.1-15.2). Four of these children had not been treated with erythropoietin before (group A), and 6 had been treated with erythropoietin administered intraperitoneally in 250 mL of dialysis fluid (group B). INTERVENTIONS: Patients in group A started on a dose of approximately 300 units/kg per week (group A). Patients in group B received their previous dose. Dosage was adjusted to achieve a target hemoglobin level of 6.5-7.0 mmol/L (104-112 g/L). Serum ferritin levels and transferrin saturation were monitored and iron supplementation was prescribed in the case of iron deficiency. MAIN OUTCOME MEASURES: Weekly erythropoietin dose in relation to hemoglobin level. RESULTS: In group A, median hemoglobin level rose from 5.3 mmol/L (85 g/L) to 6.6 mmol/L (106 g/L) after 6 months of therapy, whereas the median erythropoietin dose decreased from 266 to 234 U/kg/week. In group B, hemoglobin levels remained stable and median erythropoietin dose decreased from 262 to 194 U/kg/week. One patient in this group, for unknown reasons, never responded to erythropoietin treatment. He was excluded from further analysis. In the remaining 5 patients the median cumulative erythropoietin dose was 3250 U/kg in the 3-month period prior to the start of the study and 2713 in the 3-month period starting 6 months after the beginning of the study. This difference of 17% was statistically significant using a Wilcoxon test (p < 0.05). CONCLUSION: Intraperitoneal administration of erythropoietin in a small amount of dialysis fluid leads to a decrease in the required dose. PMID- 9358530 TI - Staphylococcus aureus carriage in adult peritoneal dialysis patients and their spouses. AB - OBJECTIVE: We hypothesized that carriage of Staphylococcus aureus among continuous ambulatory peritoneal dialysis (CAPD) patients was influenced by their spouses. Furthermore, this carrier status was compared to previous Staph. aureus peritonitis episodes in order to identify the influence of Staph. aureus carriage on peritonitis rate. DESIGN: A combined prospective surveillance study (Staph. aureus carriage) and retrospective chart review (Staph. aureus peritonitis). SETTING: A single peritoneal dialysis unit in a county hospital. PATIENTS AND METHODS: Cultures from patients (n = 32) and spouses (n = 16) were obtained twice, with a 1-month interval, from the anterior nares, the umbilical, and one groin area. All positive cultures were phage typed. Retrospective chart review of all episodes of Staph. aureus peritonitis among the patients was carried out. RESULTS: Twelve of 32 patients (37.5%) and 5 of 16 spouses (31%) evaluated were carriers. Half of the spouses of patients who were Staph. aureus carriers, were also carriers, as opposed to 20% of spouses of noncarrier patients (p = 0.30). Patients and spouses always shared the same phage type. Among patients, Staph. aureus was found in the nose only (n = 9), in all three regions (n = 2), and extranasally only (n = 1). If only one nasal culture was used to establish carriage, the sensitivity and negative predictive value would be 92% and 95%, respectively. A trend toward a higher incidence (p = 0.062) of Staph. aureus peritonitis was found among carriers (patients), 0.37 versus 0.28 peritonitis episode/dialysis-year. CONCLUSIONS: Only one positive nasal culture was necessary when carriage of Staph. aureus was to be established. Staph. aureus carriage was found more often in patients who had previously suffered from Staph. aureus peritonitis. The phage types isolated remained fairly constant, and the patients and spouses often had the same carrier state and shared the same phage types, although transmission does not always take place. PMID- 9358529 TI - Phospholipases and the activation and priming of neutrophils by peritoneal dialysis effluent. AB - OBJECTIVE: To investigate the role of phospholipase during the activation and priming of neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase by peritoneal dialysis effluent (PDE). DESIGN: Examine the action of 4-hour dwell PDE upon phospholipase activation in the circulating neutrophils obtained from healthy individuals. RESULTS: We have previously reported that PDE stimulated superoxide release by the NADPH oxidase of human neutrophils and primed the response to the bacterial peptide, fMLP (fMetLeuPhe). To elucidate the biochemical mechanisms underlying these observations, we have examined the roles of phospholipases (PL) C, D, and A2, whose activation causes the release of a range of intracellular secondary messengers. Following fMLP stimulation, we observed a rapid activation of both PLC and PLD as well as a small but nonsignificant increase in PLA2 activity. Peritoneal dialysis effluent alone failed to stimulate either PLC or PLD, while pre-incubation with PDE had no affect upon fMLP-induced PLC and PLD activation. However, PDE caused a small but nonsignificant increase in PLA2 activity (which was comparable to that observed with fMLP) and primed the fMLP-induced response. In common with a role for PLA2 and the subsequent release of arachidonic acid (AA), we have demonstrated dose dependent inhibition of PDE-induced superoxide release by the PLA2 inhibitor mepacrine, as well as activation and priming of the fMLP-induced superoxide generation by AA. CONCLUSIONS: These results imply that PDE-induced NADPH-oxidase activation and priming in human neutrophils is mediated via a PLA2-dependent but PLC- and PLD-independent mechanism. PMID- 9358531 TI - Oral versus intraperitoneal application of clindamycin in tunnel infections: a prospective, randomized study in CAPD patients. AB - OBJECTIVE: To evaluate the potential superiority of either oral or intraperitoneal treatment of catheter tunnel infections (TI), using clindamycin as a first-line antibiotic and ultrasound as a diagnostic tool. DESIGN: This was a prospective, randomized study in continuous ambulatory peritoneal dialysis patients. From August 1993 until August 1995, 16 clinically- and ultrasound proven episodes of TI were randomly assigned to either an oral or an intraperitoneal (IP) treatment (100 patients, 1414 patient-months). Main criteria for TI diagnosis were purulent drainage from the exit site and/or a positive ultrasound (pericatheter fluid collection of at least 2 mm, 7.5 MHz transducer). Initially, clindamycin (20 mg/kg body weight) was given via the oral (three times per day) or intraperitoneal route (four times per day). In the case of incompatibility or resistance to clindamycin, either oxacillin or ciprofloxacin were used orally or IP. RESULTS: Based on ultrasound criteria, the mean time until a > or = 50% reduction of pericatheter abscess diameter was 26 days (median) (range: 8-28 days) in the oral, and 15 days (8-27 days) in the IP group (p < or = 0.05). Showing no significant difference of pericatheter fluid at study entry with 4 mm (median) (range: 2-6 mm) in the oral group and 4 mm (2-4 mm) in the IP group, the IP treatment resulted in a decrease to 0 mm (0-2 mm) after 28 days (p < 0.05), while the diameter was still 2 mm (0-10 mm) (NS) in the oral group. Disappearance of exit-site infection was also somewhat earlier in the intraperitoneal group (51 vs 15 days, NS). Catheter removal had to be done once in the IP group and twice in the oral group within 6 months after study entry. CONCLUSIONS: The results give evidence for greater efficacy of the IP application of clindamycin as a first-line antibiotic compared to the oral route for the treatment of tunnel infections. PMID- 9358533 TI - Toward targets for initiation of chronic dialysis. AB - OBJECTIVES: To better define the targets for initiation of chronic dialysis, we compared the relationship between the normalized protein equivalent of nitrogen appearance (nPNA, g/kg standard weight/day) and weekly urea clearance (Kt) normalized to total body water (V) in predialysis chronic renal failure (CRF) patients and in patients on continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD). We also studied the relationships of other nutritional parameters to weekly Kt/Vurea in CRF patients. DESIGN: This cross-sectional study was a prospective observational design meant to study each patient once. SETTING: The University Hospital and Clinics and Harry S. Truman VA Medical Center, Columbia, Missouri. PATIENTS: Forty-five consecutive predialysis CRF patients were enrolled and the results compared with patients on CAPD and HD. RESULTS: In CRF, the nPNA calculated from urea appearance correlated with the weekly Kt/Vurea (r = 0.57, p < 0.0001) and, using exponential best-fit, nPNA = 1.217 x (1-e 0.769Kt/V). This exponential relationship was similar to that for CAPD and both were different from that in patients on HD. Likewise, nPNAs, calculated from Kjeldahl nitrogen output, and weekly Kt/Vurea were correlated (r = 0.37, p = 0.014) and, using exponential best-fit, nPNA = 1.102(1-e-0.867Kt/V), similar to the relationship in patients on CAPD. Evidence is presented that these relationships are not explained only by mathematical coupling. There was a significant correlation between the weekly Kt/Vurea and 24-hour urinary creatinine excretion. CONCLUSIONS: The findings suggest that in CRF, as in CAPD, a weekly Kt/Vurea less than 2.0 is likely to be associated with a nPNA less than 0.9 g/kg standard weight. In CRF patients, initiation of chronic dialysis should be considered if weekly renal Kt/Vurea falls below 2.0 and a nPNA greater than 0.8 is desired. PMID- 9358532 TI - Is heparin therapy necessary in CAPD peritonitis? AB - OBJECTIVE: Heparin therapy in continuous ambulatory peritoneal dialysis (CAPD) peritonitis seems well established; it is costly due to the necessity of hospitalization. There are no clinical studies that show a benefit of such a treatment. The aim of this study was to investigate whether heparin therapy in CAPD peritonitis is necessary. DESIGN AND PATIENTS: 194 samples of peritoneal dialysates were collected from 17 patients over a period of 24 months. Samples were subdivided into three groups: those without peritonitis (< 100 leukocytes/microL), those with mild peritonitis (100-499 leukocytes/microL), and those with severe peritonitis (> or = 500 leukocytes/microL). MEASUREMENTS: The number of leukocytes per microL dialysate and total protein concentrations were determined. Furthermore, dialysate concentrations of thrombin-antithrombin III- (TAT-) complexes (indicator of thrombin formation), D-dimers (indicator of fibrinolysis), and plasminogen activator inhibitor 1 (PAI-1) were measured. RESULTS: The dialysate protein concentration progressively increased from no peritonitis to mild and severe inflammation. In parallel, dialysate TAT-complex and D-dimer concentrations increased. Thrombin-antithrombin III-complex and D dimer concentrations correlated strongly in 179 cases (r = 0.76; 62 samples showing peritonitis, 117 samples with no evidence of peritonitis). In the remaining 15 samples of 3 patients, high PAI-1 levels (> 40 ng/mL) and low D dimer concentrations were found. Eleven of the 15 samples showed evidence of peritonitis. In these 11 samples with evidence of peritonitis, high levels of TAT complexes were detected, while D-dimer concentrations were found to be very low, pointing to a blocked fibrinolysis. The PAI-1 levels were not related to leukocyte counts or protein concentrations in the dialysates. CONCLUSIONS: Based on our findings, the routine intraperitoneal administration of heparin in CAPD peritonitis is not necessary. In rare cases an imbalance between coagulation and fibrinolysis due to high PAI-1 levels exists (15 of 194 dialysate samples, 11 of the 15 samples showing peritonitis). These cases--which do require heparinization -can be identified by demonstrating low D-dimer levels in CAPD dialysate at times of peritonitis. PMID- 9358534 TI - Foreign body granuloma at the exit site in patients on CAPD: use of 1% gentian violet as topical treatment. PMID- 9358535 TI - Increase in Kt/V increased serum albumin but not nPCR in a group of patients on continuous peritoneal dialysis. PMID- 9358536 TI - Bacillus cereus peritonitis in a chronic peritoneal dialysis patient. PMID- 9358538 TI - Literature. September-October 1997. PMID- 9358537 TI - Agrobacterium radiobacter peritonitis in a Down's syndrome child maintained on peritoneal dialysis. PMID- 9358540 TI - The determinants of physician attitudes and subjective norms toward drug information sources: modification and test of the theory of reasoned action. AB - PURPOSE: To improve upon the theory of reasoned action and apply it to pharmaceutical research, we investigated the effects of relevant appraisals attributes, and past behavior of physicians on the use of drug information sources. We also examined the moderating effects of practice characteristics. METHODS: A mail questionnaire asked HMO physicians to evaluate seven common sources of drug information on general appraisals (degree of usefulness and ease of use), specific attributes (availability, quality of information on harmful effects and on drug efficacy), and past behavior when searching for information on a new, simulated H2 antagonist agent. Semantic differential scales were used to measure each appraisal, attribute and past behavior. Information was also collected on practice characteristics. RESULTS: Findings from 108/200 respondents indicated that appraisals and attributes were useful determinants of attitudes and subjective norms toward use. Degree of usefulness and quality of information on harmful effects were important predictors of attitudes toward use for several sources of information. Ease of use and degree of usefulness were important predictors of subjective norms toward use. In many cases, moderating effects of practice characteristics were in opposing directions. Past behavior had significant direct effects on attitudes toward the PDR. CONCLUSIONS: The findings suggest ways to improve the usefulness of the theory of reasoned action as a model of decision-making. We also propose practical guidelines that can be used to improve the types of drug information sources used by physicians. PMID- 9358539 TI - Validation of a decision support system for use in drug development: pharmacokinetic data. AB - PURPOSE: Single dose pharmacokinetic data from several individuals can be used to predict the fraction of the population that is expected to be within a therapeutic range. Without having some measure of its reliability, however, that prediction is only likely to marginally influence critical drug development decision making. The system (Forecaster) described generates an approximate prediction interval that contains the original prediction and where, for example, the probability is approximately 85% that a similar prediction from a new set of data will also be within the range. The goal is to validate that the system functions as designed. METHODS: The strategy requires having a Surrogate Population (SP), which is a large number (> or = 1500) of hypothetical individuals each represented by set of model parameter values having unique attributes. The SP is generated so that a sample taken from it will give data that is statistically indistinguishable from the available experimental data. The automated method for building the SP is described. RESULTS: Validation studies using 300 independent samples document that for this example the SP can be used to make useful predictions, and that the approximate prediction interval functions as designed. CONCLUSIONS: For the boundary conditions and assumptions specified, the Forecaster can make valid predictions of pharmacokinetic-based population targets that without a SP would not be possible. Finally, the approximate prediction interval does provide a useful measure of prediction reliability. PMID- 9358541 TI - Cholera toxin B-mediated targeting of lipid vesicles containing ganglioside GM1 to mucosal epithelial cells. AB - PURPOSE: To determine whether the non-toxic pentameric B subunit of Cholera toxin (CTB) binding to ganglioside GM1 on both the lipid vesicles and epithelial cells may provide a means to target lipid vesicles to mucosal cells expressing surface GM1. METHODS: Sonicated lipid vesicles containing ganglioside GM1 were prepared. Inter-vesicle cross-linking due to pentameric CTB binding to these GM1 vesicles was determined with a sub-micron particle analyzer. Association of CTB to GM1 vesicles was analyzed with continuous sucrose gradient centrifugation. CTB mediated binding of GM1 vesicles to human mucosal epithelial cells (Caco-2 and HT 29), mucous membranes of mouse trachea, and nasal tissues were detected with fluorescent labeled vesicles. RESULTS: An increase in lipid particle size due to binding of CTB to lipid vesicles and inter-vesicles cross-linking was detected. At a 30-to-1 mole ratio of membrane-bound GM1-to-CTB, optimum increase in GM1 vesicle aggregation, was detected. Under such conditions, all the added CTB molecules were associated with GM1 vesicles. Time course analysis showed that inter-vesicles cross linking by CTB was detectable within 10 min. and reached a maximum value at 60 min. CTB associated GM1-vesicles bind to mucosal epithelial cells HT-29 and Caco-2 with similar affinity [Kd = 7.8 x 10(-4) M lipid (Caco-2) and 7.6 x 10(-4) M lipid (HT-29)]. GM1 mediated binding specificity was demonstrated by blocking with anti-GM1 antibody and the insignificant degree of CTB-associated GM1 vesicle binding to GM1 deficient C6 cells. CONCLUSIONS: The CTB-mediated GM1 binding to multiple membrane surfaces provides selective localization of GM1 vesicles to GM1 expressing mucosal cells and tissues. The strategy may be useful in localizing drugs and proteins to gut and respiratory tract mucosa. PMID- 9358542 TI - Transient expression of a purine-selective nucleoside transporter (SPNTint) in a human cell line (HeLa). AB - PURPOSE: The goal of this study was to develop a mammalian expression system for the cloned rat intestinal, Na(+)-dependent, purine-selective nucleoside transporter (SPNTint) and to study the interactions of nucleosides and nucleoside analogs with this transporter. METHODS: Lipofection was used to transfect HeLa cells with a mammalian expression vector (pcDNA3) containing the cDNA insert encoding SPNTint. Nucleoside transport activity was measured using [3H]inosine, [3H]uridine, [3H]-dideoxyinosine (ddI), and [3H]-2-chloro-2'-deoxyadenosine (2CdA) as model substrates. RESULTS: Expression of SPNTint was observed between 36 and 90 h post-transfection, with maximal expression at 66 h. At 66 h, Na(+) stimulated uptake of [3H]inosine in cells transiently transfected with SPNTint was approximately threefold greater than that in cells transfected with empty vector (p < 0.05). The Na(+)-stimulated uptake of both inosine and uridine was saturable (K(m) = 28.1 +/- 7.1 microM and 20.6 +/- 5.6 microM, respectively) in the transfected cells and was significantly inhibited by the naturally occurring nucleosides (1 mM) inosine and uridine and to a lesser extent by thymidine. The nucleoside analogs ddI (IC50 = 46 microM) and 2CdA (IC50 = 13 microM) also significantly inhibited the Na(+)-stimulated uptake of [3H]inosine. A Na(+) stimulated uptake of [3H]2CdA was observed suggesting that 2CdA is also a permeant of SPNTint. CONCLUSIONS: HeLa cells transiently transfected with SPNTint represent a useful tool to study the kinetics and interactions of drugs with SPNTint. PMID- 9358544 TI - The effect of beta-turn structure on the passive diffusion of peptides across Caco-2 cell monolayers. AB - PURPOSE: To investigate the relationships between the beta-turn structure of a peptide and its passive diffusion across Caco-2 cell monolayers, an in vitro model of the intestinal mucosa. METHODS: Linear hydrophilic peptides (Ac TyrProXaaZaaVal-NH2; Xaa = Gly, Ile and Zaa = Asp, Asn) and hydrophobic (Ac YaaPro-XaaIle Val-NH2; Yaa = Tyr, Phe and Xaa = Gly, Ile: and Ac-PhePro-XaaIle NH2; Xaa = Gly, Ile) peptides were synthesized and their effective permeability coefficients (Peff) were determined across Caco-2 cell monolayers. The lipophilicities of the peptides were estimated by measuring their partition coefficients (Po/w) between 1-octanol and HBSS. Two-dimensional NMR (2D-NMR) spectroscopy and circular dichroism (CD) spectroscopy was used to determine the solution structures of these model peptides. RESULTS: Using 2D-NMR spectroscopy and CD spectroscopy, the hydrophilic Gly-containing peptides (Ac-TyrProGlyZaaVal NH2; Zaa = Asp, Asn) were shown to exhibit a higher degree of beta-turn structure in solution than the Ile-containing peptides (Ac-TyrProIleZaaVal-NH2; Zaa = Asp, Asn). CD spectroscopy was used to show that the Gly-containing hydrophobic peptides (Ac-YaaProGlyIleVal-NH2; Yaa = Tyr, Phe: and Ac-PheProGlyIle-NH2) exhibited a higher degree of beta-turn structure in solution than the Ile containing hydrophobic peptides. The Peff values of all four hydrophilic peptides across unperturbed Caco-2 cell monolayers were very low and no statistically significant differences were observed between the Gly- and Ile-containing pentapeptides within either the Asp or Asn series. The Peff values for the hydrophobic Gly-containing peptides were significantly greater than the Peff values determined for their Ile-containing counterparts. The Gly-containing penta and tetrapeptides in the Phe series, which exhibited high permeation, were shown to be metabolically unstable. In contrast, the Gly- and Ile-containing pentapeptides in the Tyr series and the Ile-containing penta- and tetrapeptides in the Phe series, which exhibited low permeation, were metabolically stable. CONCLUSIONS: Hydrophobic peptides that exhibit significant beta-turn structure in solution are more lipophilic as measured by log Po/w and more readily permeate Caco-2 cell monolayers via the transcellular route than hydrophobic peptides that lack this type of solution structure. The ability of these peptides to permeate Caco-2 cell monolayers via the transcellular route also exposed them to metabolism, presumably by cytosolic endopeptidase. Similar secondary structural features in hydrophilic peptides do not appear to sufficiently alter the physicochemical properties of the peptides so as to alter their paracellular flux through unperturbed Caco-2 cell monolayers. PMID- 9358543 TI - Iontophoresis of poly-L-lysines: the role of molecular weight? AB - PURPOSE: (1) To determine the extent of iontophoretic transport as a function of molecular weight (MW) of the penetrant; and (2) to visually and quantitatively characterize the iontophoretic transport pathways (follicular (F) versus nonfollicular (NF) of the fluorescently-labeled poly-L-lysines employed. METHODS: A series of fluorescently-labeled poly-L-lysines (FITC-PLLs) [4 KDa, 7 KDa and 26 KDa] were used to study the extent and distribution of iontophoretic skin penetration as a function of MW using laser scanning confocal microscopy (LSCM). RESULTS: It was found that, relative to the passive controls, and under the electrical conditions considered, iontophoresis greatly enhanced the penetration of the 4 KDa analog, slightly elevated the delivery of the 7 KDa FITC-PLL, but had no effect on the transport of the larger 26 KDa FITC-PLL. Quantitative analyses of LSCM images revealed that iontophoresis increased transport via F pathways only slightly more than that through NF pathways for the 4 KDa and 7 KDa FITC-PLL molecules. CONCLUSIONS: It is visually apparent that the iontophoretic transport pathways taken are importantly determined by the physicochemical properties (including size and charge) of the penetrant. The results presented here demonstrate an inverse dependence of iontophoretic delivery upon the MW of the penetrant. PMID- 9358546 TI - Molecular modeling of polymers 16. Gaseous diffusion in polymers: a quantitative structure-property relationship (QSPR) analysis. AB - PURPOSE: The purpose of this study was to identify the key physicochemical molecular properties of polymeric materials responsible for gaseous diffusion in the polymers. METHODS: Quantitative structure-property relationships, QSPRs were constructed using a genetic algorithm on a training set of 16 polymers for which CO2, N2, O2 diffusion constants were measured. Nine physicochemical properties of each of the polymers were used in the trial basis set for QSPR model construction. The linear cross-correlation matrices were constructed and investigated for colinearity among the members of the training sets. Common water diffusion measures for a limited training set of six polymers was used to construct a "semi-QSPR" model. RESULTS: The bulk modulus of the polymer was overwhelmingly found to be the dominant physicochemical polymer property that governs CO2, N2 and O2 diffusion. Some secondary physicochemical properties controlling diffusion, including conformational entropy, were also identified as correlation descriptors. Very significant QSPR diffusion models were constructed for all three gases. Cohesive energy was identified as the main correlation physicochemical property with aqueous diffusion measures. CONCLUSIONS: The dominant role of polymer bulk modulus on gaseous diffusion makes it difficult to develop criteria for selective transport of gases through polymers. Moreover, high bulk moduli are predicted to be necessary for effective gas barrier materials. This property requirement may limit the processing and packaging features of the material. Aqueous diffusion in polymers may occur by a different mechanism than gaseous diffusion since bulk modulus does not correlate with aqueous diffusion, but rather cohesive energy of the polymer. PMID- 9358545 TI - The effect of beta-turn structure on the permeation of peptides across monolayers of bovine brain microvessel endothelial cells. AB - PURPOSE: To investigate the effects of the beta-turn structure of a peptide on its permeation via the paracellular and transcellular routes across cultured bovine brain microvessel endothelial cell (BBMEC) monolayers, an in vitro model of the blood-brain barrier (BBB). METHODS: The effective permeability coefficients (Peff) of the model peptides were determined across BBMEC monolayers. The dimensions of the aqueous pores in the tight junctions (TJs) of the BBMEC monolayers were determined using a series of hydrophilic permeants. This value and the molecular radius of each peptide were used to calculate the theoretical paracellular (PP*) and transcellular (PT*) permeability coefficients for each peptide. RESULTS: A comparison of the theoretical PP* values with the observed Peff values was made for a series of model peptides. For the most hydrophobic peptides (Ac-PheProXaaIle-NH2 and Ac-PheProXaaIleVal-NH2; Xaa = Gly, Ile), it was concluded that the Gly-containing peptide of each pair more readily permeates BBMEC monolayers via the transcellular pathway than the Ile-containing analog. In addition, the Gly-containing peptides, which exhibit more beta-turn structure, were shown to be more lipophilic than the Ile-containing peptides as estimated by the log of their 1-octanol:HBSS partition coefficients (log Po/w). However, the three hydrophilic peptide pairs (Ac-TyrProXaaAspVal-NH2, Ac TyrProXaaAsnVal-NH2, and Ac-TyrProXaaIleVal-NH2; Xaa = Gly, Ile) were found to permeate BBMEC monolayers predominantly via the paracellular pathway. No differences were observed in the Peff values of the hydrophilic peptides having higher beta-turn structures as compared to the peptides lacking these structural features. In addition, the Ile-containing peptides exhibited significantly higher log Po/w values than the Gly-containing hydrophilic peptides. CONCLUSIONS: Hydrophobic peptides that exhibit significant beta-turn structure in solution are more lipophilic as measured by log Po/w, and more readily permeate BBMEC monolayers via the transcellular route than hydrophobic peptides that lack this type of solution structure. Similar secondary structural features in hydrophilic peptides do not appear to sufficiently alter the physicochemical properties of the peptides so as to alter their paracellular flux through BBMEC monolayers. PMID- 9358547 TI - On the variability of dissolution data. AB - PURPOSE: To investigate dissolution data variability and its origins. METHODS: The Weibull function with four parameters t0 (dissolution lag-time), K (the rate parameter), beta (the shape parameter) and D (the fraction dissolved as t- >infinity), is used to describe the dissolution curve. The variance of the dissolution data is expressed in terms of these parameters and their individual variances sigma t02, sigma K2, sigma beta 2, and sigma D2. These four variances originate from variable physical properties of the dosage units and from a variable dissolution environment. Therefore, dissolution data variability depends on both, the functional form of the curve and on the variance of the physical conditions. The use of this method enables the elucidation of the sources of dissolution data variability. RESULTS: In the case of a sigmoidal dissolution curve (beta > 1), data variance is zero as dissolution begins (following dissolution lag-time). This initial variance diverges when the dissolution curve is non-sigmoidal (with beta < 1) but assumes a finite value, proportional to the dissolution lag-time variance (sigma t02) when the data fits a regular first order rate curve (beta = 1). Following a long dissolution time, data variance attains a constant value equal to the dissolution extent variance, sigma D2. When the dissolution curve is sigmoidal and the variability related to the dissolution extent is sufficiently small (sigma D/D < < 1), a maximum in the variance is expected at some intermediate time point (corresponding to the curve inflection point, when the main source of variability is dissolution lag-time t0, or around t = 1/K + t0, when the main sources of variability are the rate parameter K or the shape parameter beta). When the curve is sigmoidal (beta > 1) and the main source of variability relates to the dissolution extent, the overall variance grows with time all the way to the plateau of the dissolution curve. With a non sigmoidal dissolution curve (beta < or = 1), data variability decreases with time soon after dissolution begins. In that case, if the main source of variability is the dissolution lag-time (t0), the variance decreases all the way to the plateau of the dissolution curve. If the dissolution extent, D, is the main source of variability, a minimum in the variance is expected at some intermediate time point. The dissolution relative variance, on the other hand, diverges as dissolution begins and decreases with time at least until 63% of the drug is released, irrespective to the Weibull parameter values. Later, it may decrease or increase, attaining a fixed value (sigma D2/D2) at the plateau of the dissolution curve. CONCLUSIONS: The particular time dependence of dissolution data variance is well defined in terms of the Weibull shape parameters and their individual variances. Dissolution data variability may decrease or increase with time long the curve. It may attain a maximum or a minimum value at some intermediate time point. It may converge or diverge as dissolution begins. When the dissolution data is well fitted to the Weibull function, the sources of data variability (in terms of the Weibull parameters) may be elucidated. The variability of dissolution data originates from physical sources but is also dependent on the functional form of the curve. PMID- 9358548 TI - Evaluation of direct curve comparison metrics applied to pharmacokinetic profiles and relative bioavailability and bioequivalence. AB - PURPOSE: The intent was to investigate three direct curve comparison metrics, the Rescigno Index, f1, and the Chinchilli Metric as tools to assess relative bioavailability (BA) and bioequivalence (BE). The specific objectives were to 1) estimate relative bioavailability and bioequivalence and 2) compare detection of profile shape differences with typical (i.e. AUC and Cmax) criteria. METHODS: Product bioequivalence was estimated and the degree of concordance with typical criteria in detecting profile differences was determined from the eighteen bioequivalence studies (390 subjects). Descriptive statistical analysis was carried out on the concordant and discordant profile subsets. RESULTS: 1) Three of the eighteen studies failed typical criteria (AUC and Cmax). Of the curve metrics, 12 studies failed the Chinchilli metric criteria and 13 failed f1 criteria. Using three different exponents in the Rescigno Index calculation, 17 (exponent = 3), 13 (exponent = 1), and 11 (exponent = 1/3) failed the criteria for bioequivalence. The frequency of profiles found to be different was comparable across the metrics, but the specific profiles found to be different or not different varied across the metrics. The Chinchilli Metric and f1 agreed 71% and 72% with typical criteria in judging profiles to be different or not different. Descriptive evaluation suggested that the direct curve metrics more effectively detect differences in absorption time lags but less effectively detect differences in Cmax. The Rescigno Index showed dependence on the direction of the difference between test and reference concentrations. CONCLUSIONS: With the limits used here, the direct curve metrics represent a more conservative approach to evaluate product bioequivalence. With further investigation and development, the direct curve approach may be used effectively to evaluate relative BA and BE. PMID- 9358549 TI - Long-term and high-temperature storage of supercritically-processed microparticulate protein powders. AB - PURPOSE: The long-term and high-temperature storage of dry, micron-sized particles of lysozyme, trypsin, and insulin was investigated. Subsequent to using supercritical carbon dioxide as an antisolvent to induce their precipitation from a dimethylsulfoxide solution, protein microparticles were stored in sealed containers at -25, -15, 0, 3, 20, 22, and 60 degrees C. The purpose of this study was to investigate the suitability of supercritical antisolvent precipitation as a finishing step in protein processing. METHODS: Karl Fisher titrations were used to determine the residual moisture content of commercial and supercritically processed protein powders. The secondary structure of the dry protein particles was determined periodically during storage using Raman spectroscopy. The proteins were also redissolved periodically in aqueous buffers and assayed spectrophotometrically for biological activity and by circular dichroism for structural conformation in solution. RESULTS: Amide I band Raman spectra indicate that the secondary structure of the protein particles, while perturbed from that of the solution state, remained constant in time, regardless of the storage temperature. The recoverable biological activity upon reconstitution for the supercritically-processed lysozyme and trypsin microparticles was also preserved and found to be independent of storage temperature. Far UV circular dichroism spectra support the bioactivity assays and further suggest that adverse structural changes, with potential to hinder renaturation upon redissolution, do not take place during storage. CONCLUSIONS: The present study suggests that protein precipitation using supercritical fluids may yield particles suitable for long-term storage at ambient conditions. PMID- 9358551 TI - Interaction of human IgE with soluble forms of IgE high affinity receptors. AB - PURPOSE: Interaction of human IgE with its high affinity receptor (Fc epsilon RI) on mast cells and basophils is an important step for initiating IgE mediated immune responses. To characterize the IgE and Fc epsilon RI interaction, we investigated this interaction in terms of stoichiometry and binding affinity in solution. The binding of IgE and IgE Fc epsilon RI alpha chain, the extracellular portion of IgE high affinity receptor (sFc epsilon RI alpha) was compared with the binding of IgE and IgE immunoadhesin (Fc epsilon RI alpha-IgG). METHODS: The interaction was characterized by analytical ultracentrifugation, size exclusion chromatography, light scattering and ELISA. RESULTS: We show that the sFc epsilon RI alpha is only able to bind to one IgE, while the immunoadhesin can bind to two IgE. The interaction between IgE and Fc epsilon RI is very strong. Both forms of soluble receptors have similar intrinsic binding affinity with IgE. CONCLUSIONS: Both soluble receptors (Fc epsilon RI alpha-IgG and sFc epsilon RI alpha) can block the binding of IgE to its high affinity receptors on cell surface. The Fc epsilon RI alpha-IgG is a better IgE binding protein than sFc epsilon RI alpha at physiological relevant conditions. A humanized anti-IgE monoclonal antibody, rhuMAb E25 that also can block the binding of IgE to its high affinity receptors appears to bind to IgE at slightly different regions or in a different manner as the soluble forms of IgE receptors. PMID- 9358550 TI - The stability of insulin in crystalline and amorphous solids: observation of greater stability for the amorphous form. AB - PURPOSE: Generalizations based upon behavior of small molecules have established that a crystalline solid is generally much more stable toward chemical degradation than is the amorphous solid. This study examines the validity of this generalization for proteins using biosynthetic human insulin as the model protein. METHODS: Amorphous insulin was prepared by freeze drying the supernate from a suspension of zinc insulin crystals adjusted to pH 7.1. Storage stability at 25 degrees C and 40 degrees C were compared for the freeze dried material, the dried suspended crystals, and the starting batch of crystals. Samples were equilibrated at selected relative humidities between zero and 75% to obtain samples at various water contents. Assays for dimer formation were performed by size exclusion HPLC and assays for deamidated product were carried out by reverse phase HPLC. Degradation was found to be linear in square root of time, and the slopes from % degradation vs. square root of time were used to define the rate constants for degradation. Differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR) were used to characterize the state of the protein in the solids. RESULTS: As expected based upon previous results, the primary degradation pathways involve deamidation at the AsnA21 site and co-valent dimer formation, presumably involving the A-21 site. Contrary to expectations, amorphous insulin is far more stable than crystalline insulin under all conditions investigated. While increasing water content increases the rate of degradation of crystalline insulin, rate constants for degradation in the amorphous solid are essentially independent of water content up to the maximum water content studied (approximately 15%). CONCLUSIONS: Based upon the FTIR and DSC data, both crystalline and amorphous insulin retain some higher order structure when dried, but the secondary structure is significantly perturbed from that characteristic of the native solution state. However, neither DSC nor FTIR data provide a clear interpretation of the difference in stability between the amorphous and crystalline solids. The mechanism responsible for the superior stability of amorphous insulin remains obscure. PMID- 9358552 TI - Development and use of enzyme-linked immunosorbent assays (ELISA) for the detection of protein aggregates in interferon-alpha (IFN-alpha) formulations. AB - PURPOSE: Protein aggregates are thought to be involved in the immunogenicity of recombinant proteins in humans. To probe human IFN-alpha formulations for the presence of soluble protein aggregates, enzyme-linked immunosorbent assays (ELISA) were developed. METHODS: For the detection of IFN-alpha-IFN-alpha and HSA IFN-alpha aggregates, sandwich ELISAs were developed using a monoclonal anti-IFN alpha antibody as a capture antibody and the same anti-IFN-alpha antibody and an anti-human serum albumin (HSA) antibody (HRP-labeled), respectively. RESULTS: Marketed freeze-dried, HSA-containing IFN-alpha-formulations tested in the ELISAs all contained IFN-alpha-IFN-alpha and/or HSA-IFN-alpha protein aggregates, although in varying amounts. These aggregates were predominantly IFN-alpha dimers and 1:1 conjugates of HSA with IFN-alpha. Test formulations revealed that aggregation of IFN-alpha was strongly affected by the presence of pharmaceutical excipients, pH of the formulation, lyophilisation procedure, and storage temperature and time. CONCLUSIONS: The ELISAs are rapid, highly specific for aggregates in the presence of both IFN-alpha and HSA monomers and allow the direct detection of both types of aggregates in formulations in the nanogram range. The new assays will assist the monitoring of the aggregate-inducing processes during IFN-alpha formulation and storage in an early phase and the development of aggregate-free IFN-alpha formulations. PMID- 9358553 TI - Protection afforded by maltosyl-beta-cyclodextrin against alpha-chymotrypsin catalyzed hydrolysis of a luteinizing hormone-releasing hormone agonist, buserelin acetate. AB - PURPOSE: The present study addresses how maltosyl-beta-cyclodextrin (G2-beta-CyD) impacts upon the alpha-chymotrypsin-catalyzed hydrolysis of buserelin acetate, an agonist of luteinizing hormone-releasing hormone with emphasis upon the direct effect of G2-beta-CyD on the activity of the protease. METHODS: Kinetic and solubility studies were performed in isotonic phosphate buffer (pH 7.4) at 25 degrees C and 37 degrees C. The interaction of alpha-chymotrypsin with G2-beta CyD in the buffer solution was examined by differential scanning calorimetry. RESULTS: G2-beta-CyD decelerated the alpha-chymotrypsin-catalyzed hydrolysis of buserelin acetate to give the 1-3 tripeptide and the 4-9 hexapeptide fragments. This deceleration can be explained solely by a non-productive encounter between a complex of the substrate with G2-beta-CyD and the protease at relatively low CyD concentrations, while the direct inhibitory effect of G2-beta-CyD on the proteolytic activity made a considerable contribution to the overall deceleration of the hydrolysis at higher CyD concentrations. Calorimetric studies indicate the presence of intermediate states in the thermal unfolding of alpha-chymotrypsin, simultaneously accompanied by the autolysis. By contrast, a two-state thermal unfolding of alpha-chymotrypsin was observed in the presence of G2-beta-CyD, suggesting reduced proteolytic activity upon binding to G2-beta-CyD. CONCLUSIONS: These results suggest that G2-beta-CyD at higher concentrations inhibits the proteolytic action of alpha-chymotrypsin through direct interaction with the protease, as well as through the formation of a non-productive complex with the substrate. PMID- 9358554 TI - Stabilization of 10-hydroxycamptothecin in poly(lactide-co-glycolide) microsphere delivery vehicles. AB - PURPOSE: The purpose of this study was to investigate the potential of poly(lactide-co-glycolide) (PLGA) microspheres to stabilize and deliver the analogue of camptothecin, 10-hydroxycamptothecin (10-HCPT). METHODS: 10-HCPT was encapsulated in PLGA 50:50 microspheres by using an oil-in-water emulsion-solvent evaporation method. The influence of encapsulation conditions (i.e., polymer molecular weight (Mw), polymer concentration, and carrier solvent composition) on the release of 10-HCPT from microspheres at 37 degrees C under perfect sink conditions was examined. Analysis of the drug stability in the microspheres was performed by two methods: i) by extraction of 10-HCPT from microspheres and ii) by sampling release media before lactone--carboxylate conversion could take place. RESULTS: Microspheres made of low Mw polymer (inherent viscosity 0.15 dl/g) exhibited more continuous drug release than those prepared from polymers of higher Mw (i.v. = 0.58 and 1.07 dl/g). In addition, a high polymer concentration and the presence of cosolvent in the carrier solution to dissolve 10-HCPT were both necessary in the microsphere preparation in order to eliminate a large initial burst of the released 10-HCPT. An optimal microsphere formulation released 10-HCPT slowly and continuously for over two months with a relatively small initial burst of the released drug. Both analytical methods used to assess the stability of 10-HCPT revealed that the unreleased camptothecin analogue in the microspheres remained in its active lactone form (> 95%) over the entire 2 month duration of study. CONCLUSIONS: PLGA carriers such as those described here may be clinically useful to stabilize and deliver camptothecins for the treatment of cancer. PMID- 9358555 TI - A new ternary polymeric matrix system for controlled drug delivery of highly soluble drugs: I. Diltiazem hydrochloride. AB - PURPOSE: The purpose of this study was to develop a new ternary polymeric matrix system that is easy to manufacture and that delivers a highly soluble drug over long periods of time. METHODS: Pectin, hydroxypropylmethylcellulose (HPMC), and diltiazem HCl granulated with gelatin at optimized ratios were blended at different loading doses and directly compressed. Swelling behavior, dissolution profiles and the effect of hydrodynamic stress on release kinetics were evaluated. RESULTS: Diltiazem release kinetics from the ternary polymeric system was dependent on the different swelling behavior of the polymers and varied with the drug loading dose and hydrodynamic conditions. Drug release followed either non-Fickian or Case II transport kinetics. The relative influence of diffusion and relaxational/dissolution effects on release profiles for different drug loadings was calculated by a nonlinear regression approach. Photographs taken during swelling show that the anisotropic nature of the gel structure, drug loading dose, swelling capacity of polymers used, and the design of delivery system all play important roles in controlling the drug release and dissolution/erosion processes. CONCLUSIONS: Zero-order delivery of diltiazem HCl from a simple tablet matrix was achieved. The ternary polymeric system developed in this study is suitable for controlled release of highly soluble drugs. It offers a number of advantages over existing systems, including ease of manufacturing and of release modulation, as well as reproducibility of release profiles under well defined hydrodynamic conditions. Our delivery system has the potential to fully release its drug content in a controlled manner over a long time period and to dissolve completely. PMID- 9358556 TI - Characterization of poly(glycolide-co-D,L-lactide)/poly(D,L-lactide) microspheres for controlled release of GM-CSF. AB - PURPOSE: This study describes the preparation and characterization of a controlled release formulation of granulocyte-macrophage colony-stimulating factor (GM-CSF) encapsulated in poly(glycolide-co-D,L-lactide) (PLGA) and poly(D,L-lactide) (PLA) microspheres. METHODS: GM-CSF was encapsulated in PLGA/PLA microspheres by a novel silicone oil based phase separation process. Several different blends of PLGA and low molecular weight PLA were used to prepare the microspheres. The microspheres and the encapsulated GM-CSF were extensively characterized both in vitro and in vivo. RESULTS: Steady release of GM-CSF was achieved over a period of about one week without significant "burst" of protein from the microspheres. Analysis of microsphere degradation kinetics by gel permeation chromatography (GPC) indicated that low molecular weight PLA enhanced the degradation of the PLGA and thereby affected release kinetics. GM CSF released from the microspheres was found to be biologically active and physically intact by bioassay and chromatographic analysis. Analysis of serum from mice receiving huGM-CSF indicated that the GM-CSF was biologically active and that a concentration of greater than 10 ng/mL was maintained for a period lasting at least nine days. MuGM-CSF was not detected following in vivo administration of muGM-CSF microspheres. The tissues of mice receiving muGM-CSF microspheres were characterized by infiltration of neutrophils, and macrophages which were in significant excess of those found in mice administered with placebo controls (i.e. microspheres without GM-CSF). CONCLUSIONS: This study demonstrates the influence of formulation parameters on the encapsulation of GM-CSF in PLGA/PLA microspheres and its controlled release in biologically active form. The intense local tissue reaction in mice to muGM-CSF microspheres demonstrates the importance of the mode of delivery on the pharmacologic activity of GM-CSF. PMID- 9358558 TI - Effect of drug load and plate coating on the particle size distribution of a commercial albuterol metered dose inhaler (MDI) determined using the Andersen and Marple-Miller cascade impactors. AB - PURPOSE: The purpose of this study is to investigate the effect of drug load, the coating of impactor stages, and the design of cascade impactors on albuterol MDIs particle size distribution measurements. The results of the investigation will be used to explain the "loading effect" recently reported. METHODS: Particle size distribution parameters of a commercial albuterol MDI were measured using both Andersen (AI) and Marple-Miller (MMI) Cascade Impactors, where plates were either left uncoated or coated with silicone or glycerin. A previously validated HPLC-EC method was used for the assay of albuterol collected by the impactor and in single spray content determinations. RESULTS: Coating impactor collection plates had an impact on measured MMAD and GSD values for single puff measurements but very little or no effect for the multi puff measurements. Due to particle bounce, the percent of albuterol fine particles deposited in the filter and impactor finer stages (< 1.10 microns in AI and < 1.25 microns in MMI) in uncoated single puff experiments was much higher in comparison to either coated single puff or multi-puff (coated and uncoated) measurements. CONCLUSIONS: Evaluation of drug load and plate coating are necessary to determine whether observed particle size distributions are representative of the generated aerosol or are the result of particle bounce and reentrainment. In order to minimize particle bounce, especially for single puff determinations, it may be useful to apply a thin layer of a sticky coating agent to the surfaces of impactor plates. PMID- 9358557 TI - Chitosan and chitosan/ethylene oxide-propylene oxide block copolymer nanoparticles as novel carriers for proteins and vaccines. AB - PURPOSE: The aim of this study was to investigate the interaction between the components of novel chitosan (CS) and CS/ethylene oxide-propylene oxide block copolymer (PEO-PPO) nanoparticles and to evaluate their potential for the association and controlled release of proteins and vaccines. METHODS: The presence of PEO-PPO on the surface of the nanoparticles and its interaction with the CS was identified by X-ray photoelectron spectroscopy (XPS). The mechanism of protein association was elucidated using several proteins, bovine serum albumin (BSA), and tetanus and diphtheria toxoids, and varying the formulation conditions (different pH values and concentrations of PEO-PPO), and the stage of protein incorporation into the nanoparticles formation medium. RESULTS: BSA and tetanus and diphtheria toxoids were highly associated with CS nanoparticles partly due to electrostatic interactions between the carboxyl groups of the protein and the amine groups of CS. PEO-PPO also interacted electrostatically with CS, thus competing with the proteins for association with CS nanoparticles. A visible amount of PEO-PPO was projected towards the outer phase of the nanoparticles. Proteins were released from the nanoparticles at an almost constant rate, the intensity of which was closely related to the protein loading. Furthermore, the tetanus vaccine was released in the active form for at least 15 days. CONCLUSIONS: CS and CS/PEO-PPO nanoparticles prepared by a very mild ionic crosslinking technique are novel and suitable systems for the entrapment and controlled release of proteins and vaccines. PMID- 9358559 TI - Glucuronidation of entacapone, nitecapone, tolcapone, and some other nitrocatechols by rat liver microsomes. AB - PURPOSE: Nitrocatechol COMT inhibitors are a new class of bioactive compounds, for which glucuronidation is the most important metabolic pathway. The objective was to characterize the enzyme kinetics of nitrocatechol glucuronidation to improve the understanding and predicting of the pharmacokinetic behavior of this class of compounds. METHODS: The glucuronidation kinetics of seven nitrocatechols and 4-nitrophenol, the reference substrate for phenol UDP-glucuronosyltransferase activity, was measured in liver microsomes from creosote-treated rats and determined by non-linear fitting of the experimental data to the Michaelis-Menten equation. A new method that combined densitometric and radioactivity measurement of the glucuronides separated by HPTLC was developed for the quantification. RESULTS: Apparent K(m) values for the nitrocatechols varied greatly depending on substitution pattern being comparable with 4-nitrophenol (0.11 mM) only in the case of 4-nitrocatechol (0.19 mM). Simple nitrocatechols showed two-fold Vmax values compared with 4-nitrophenol (68.6 nmol min-1 mg-1), while all disubstituted catechols exhibited much lower glucuronidation rate. Vmax/K(m) values were about 10 times higher for monosubstituted catechols compared to disubstituted ones. The kinetic parameters for COMT inhibitors were in the following order: K(m) nitecapone > > entacapone > tolcapone; Vmax nitecapone > entacapone > tolcapone; Vmax/K(m) tolcapone > nitecapone > entacapone. CONCLUSIONS: Nitrocatechols can in principle be good substrates of UGTs. However, substituents may have a remarkable effect on the enzyme kinetic parameters. The different behaviour of nitecapone compared to the other COMT inhibitors may be due to its hydrophilic 5-substituent. The longer elimination half-life of tolcapone in vivo compared to entacapone could not be explained by glucuronidation kinetics in vitro. PMID- 9358560 TI - Biomimetic metabolism of artelinic acid by chemical cytochrome P-450 model systems. AB - PURPOSE: To study the reaction of artelinic acid with chemical model systems of cytochrome P-450 as a means of obtaining authentic samples of the putative metabolites necessary for identification of the mammalian metabolites of artelinic acid. METHODS: Artelinic acid was reacted with different organic complexes of iron(II). The reaction products were isolated and characterized by NMR and thermospray mass spectroscopy. RESULTS: Five compounds which are putative metabolites of artelinic acid were isolated from these reactions and unambiguously identified, while the identity of two other compounds await final confirmation. CONCLUSIONS: Standards of possible metabolites of artelinic acid can be produced by the reaction of the compound with ferrous complexes that may simulate cytochrome P-450 catalyzed metabolism of xenobiotics. This approach may provide a simple and versatile method for the formation of metabolites of artemisinin compounds which is more advantageous than previous approaches with fungal-based systems. PMID- 9358561 TI - The design and validation of a novel intravenous microdialysis probe: application to fluconazole pharmacokinetics in the freely-moving rat model. AB - PURPOSE: The purpose of this study was to design and validate a concentric, flexible intravenous microdialysis probe to determine drug concentrations in blood from the inferior vena cava of a freely-moving animal model. METHODS: An intravenous microdialysis probe was constructed using fused-silica tubing and an acrylonitrile/sodium methallyl sulfonate copolymer hollow fiber. The probe was tested in vitro for the recovery of fluconazole and UK-54,373, a fluconazole analog used for probe calibration by retrodialysis. Subsequent in vivo validation was done in rats (n = 7) that had a microdialysis probe inserted into the inferior vena cava via the femoral vein, and the femoral artery was cannulated for simultaneous blood sampling. Comparisons of fluconazole pharmacokinetic parameters resulting from the two sampling methods were performed at 2 and 10 days after probe implantation. RESULTS: There were no statistical differences between the microdialysis sampling and conventional blood sampling methods for the T1/2, Cl, Vdss, and dose-normalized AUC by paired t-test (p > 0.05) for repeated dosing at day 2 and day 10 after probe placement. The probe recovery, as determined by retrodialysis, significantly decreased over the ten day period. This finding indicates the necessity for frequent recovery determinations during a long-term blood microdialysis experiment. CONCLUSIONS: These results show that microdialysis sampling in the inferior vena cava using this unique and robust probe design provides an accurate method of determining blood pharmacokinetics in the freely-moving rat for extended experimental periods. The probe design allows for a simple surgical placement into the inferior vena cava which results in a more stable animal preparation for long-term sampling and repeated-measures experimental designs. PMID- 9358562 TI - The pharmacokinetics of topotecan and its carboxylate form following separate intravenous administration to the dog. AB - PURPOSE: To determine the relationship between topotecan and its ring opened hydrolysis product (SK&F 105,992) following intravenous administration of the two agents separately, and to determine the bio-availability of topotecan in female beagle dogs. METHODS: The pharmacokinetics of topotecan and SK&F 105,992 were determined following separate administration as 30 minute intravenous infusions in a cross-over design. Topotecan was also administered orally to the same dogs. RESULTS: When administered intravenously to dogs, SK&F 105,992 underwent interconversion to topotecan. Plasma concentrations of both topotecan and SK&F 105,992 appeared to decline multi-exponentially following i.v. infusion of either compound. A 2-compartment model was found to adequately characterize the data. CONCLUSIONS: The clearance of topotecan by other routes proceeded at a faster rate than its interconversion to SK&F 105,992, whereas the clearance of SK&F 105,992 by other routes was slower than the rate of its interconversion to topotecan. Any SK&F 105,992 formed in the GI tract did not appear to be well absorbed following oral administration of topotecan to dogs. The steady-state volume of distribution for topotecan was approximately 8- to 9-fold greater than that for SK&F 105,992 in the dog. After intravenous administration of topotecan, the amount of topotecan in the dog was much greater than that of the carboxylate, even though their respective plasma concentrations were similar. The bioavailability of topotecan, calculated from oral topotecan data or from SK&F 105,992 data, was approximately 50%. PMID- 9358564 TI - Protein aggregates seem to play a key role among the parameters influencing the antigenicity of interferon alpha (IFN-alpha) in normal and transgenic mice. AB - PURPOSE: During long-term treatment of various malignant or viral diseases with IFN-alpha up to 20% of patients develop anti-IFN-alpha antibodies for as yet unknown reasons. METHODS: To address this issue, a mouse model using Balb/C mice was established and the relevance of several potentially anti-IFN-alpha antibodies inducing factors was studied. RESULTS: The model revealed that both a higher frequency of injections and a higher dosage of IFN-alpha were more immunogenic and that the route of administration affected the antibody response to IFN-alpha. The intrinsic immunostimulatory activity of IFN-alpha itself also enhanced the immune response. IFN-alpha protein aggregates (IFN-alpha-IFN-alpha and human serum albumin (HSA)-IFN-alpha aggregates), which were recently identified in all marketed IFN-alpha products, were significantly more immunogenic than IFN-alpha monomers. These aggregates broke the tolerance against human IFN-alpha monomers in human IFN-alpha transgenic mice. CONCLUSIONS: Based on these animal studies it is proposed that the immune response to IFN-alpha in humans is most probably elicited by a combination of several factors among which IFN-alpha protein aggregates seem to play a key role. PMID- 9358563 TI - Effects of low and high density lipoproteins on renal cyclosporine A and cyclosporine G disposition in the isolated perfused rat kidney. AB - PURPOSE: This study investigated the effects of low (LDL) and high density lipoproteins (HDL) on renal cyclosporine A (CsA) and cyclosporine G (CsG) disposition in the isolated perfused rat kidney model. METHODS: Kidneys were perfused with CsA or CsG in perfusion medium containing 6% protein, bovine serum albumin only (BSA) (Control), LDL (200 mg/dl) and BSA, or HDL (200 mg/dl) and BSA. In vitro protein binding studies were conducted with CsA and CsG in the same media. RESULTS: The unbound fractions (fu) of CsA and CsG were significantly reduced with LDL and HDL in the perfusion media. In the presence of LDL, fu for CsA and CsG was 3.9% and 5.9%, respectively. With HDL, fu was 2.1% for CsA and 1.8% for CsG. fu for the controls was 14.7% for CsA and 11.9% for CsG. Renal clearance (CLR) of CsA and CsG was significantly reduced when perfused with perfusion medium containing LDL and HDL. LDL and HDL had similar effects on reducing CsA and CsG CLR, and were approximately four-fold lower when compared to controls (approximately 0.006 Vs. 0.023 ml/min). Renal CsA and CsG tissue (whole organ, cortex and medulla) concentrations were lower than corresponding controls when perfused with LDL or HDL. CONCLUSIONS: The interaction of CsA and CsG with LDL and HDL significantly reduced the CLR and extent of renal tissue distribution of both compounds. PMID- 9358565 TI - Lymphatic uptake and biodistribution of liposomes after subcutaneous injection: III. Influence of surface modification with poly(ethyleneglycol). AB - PURPOSE: The aim of the present paper was to assess the effect of inclusion of distearoylphosphatidylethanolamine-poly(ethyleneglycol) (DSPE-PEG) into liposomal bilayers on the lymphatic uptake and lymph node localization of liposomes after subcutaneous administration. METHODS: [3H]-Cholesteryloleylether labeled liposomes of various composition and sizes were injected s.c. into the dorsal side of the foot of rats. At several time-points after injection, blood levels of liposomes were determined. Lymphatic uptake from the s.c. site of injection and lymph node localization in regional lymph nodes were determined at the end of the 52 h observation period. RESULTS: The results demonstrate that inclusion of DSPE PEG into several types of liposomes has only a modest effect on lymphatic uptake. Also lymph node localization is only slightly affected by PEG-mediated steric stabilization. CONCLUSIONS: Factors other than the presence of a steric barrier are more important in determining lymphatic uptake from the s.c. injection site. The observation that lymph node localization was only slightly affected by PEG coating strongly suggests that macrophage uptake is not the only important mechanism of lymph node localization of s.c. administered liposomes. PMID- 9358567 TI - A new approach for direct in vivo dissolution studies of poorly soluble drugs. PMID- 9358566 TI - Relationship between size of injected microspheres and ultrasound imaging of heart by PLSR. AB - PURPOSE: To investigate the enhancement in ultrasound signal (US) in heart, for diagnostic purposes, as a function of size and number of injected air-filled microspheres by multivariate statistical methods. METHODS: The US enhancement in left ventricle was measured after injection of 31 suspensions of air-filled albumin microspheres divided on and injected in six dogs. The relationship between the size of microspheres between 1-38 microns and the US enhancement was explored by Pearson's product moment correlation analysis, ordinary least-squares regression, principal component regression (PCR) and partial least-squares regression (PLS). RESULTS: Relative advanced algorithms such as PCR and PLS were required to achieve accurate in vivo/in vitro correlation. The most effective microsphere sizes contributing to US enhancement in left ventricle in dogs were estimated to be in the 7-15 microns range. CONCLUSIONS: In general, the effective in vivo sizes are dependent on the type of formulation due to the surprisingly large active in vivo sizes found for the tested concept. PCR and PLS are suitable methods for in vivo/in vitro correlation, especially for covariated and noisy data. PMID- 9358568 TI - Accuracy of routine ultrasonography in screening heart disease prenatally. Gruppo Piemontese for Prenatal Screening of Congenital Heart Disease. AB - The aim of the present study was to assess the accuracy of the four-chamber view as a screening test for detection of congenital heart disease (CHD) prenatally in a low-risk population. A prospective observational study was conducted in 17 ultrasound units of the Piemonte Region, Italy, in pregnancies with no risk factors for CHD. At each routine scan, from 18 weeks of gestational age, the four chamber view of the heart was looked for. When an anomaly was suspected, the patients were referred to a specialized unit. Follow-up of the babies until discharge from the hospital was obtained. 11,232 sonograms were performed on 8299 pregnancies. Cardiac malformations were diagnosed in 40 newborns (4.8/1000). Six of them (15 per cent) had been recognized in utero. The sensitivity, specificity, and positive and negative predictive values were 15, 99.9, 50, and 99.6 per cent, respectively. When malformations that are not associated with an abnormal four chamber view were excluded from the analysis, the sensitivity increased to 35.3 per cent. The sensitivity found in this study is low, but it is probably realistic since it is comparable to that reported in other multicentric studies. This type of study should reflect the state of the art of the method applied in the field. Although the sensitivity is low, it would be nil if the test were not performed. Moreover, it will probably increase with better training of the operators and by extending the examination to the ventriculo-arterial connections. PMID- 9358569 TI - Prenatal sonographic diagnosis of vermal agenesis. AB - Agenesis of the vermis as detected during gestation by ultrasonography may indicate the existence of various malformation arrays or syndromes. We report on our observations of five cases of complete vermal agenesis that were detected at 22-31 weeks of gestation. All had a vertex presentation and transvaginal sonography established the diagnosis of vermal agenesis. Two of the vermal agenesis cases had no associated anomalies outside the central nervous system (CNS). In one, the cerebellar cleft was the only abnormality present and the other also had lobar holoprosencephaly. The three remaining fetuses had trisomy 13 and featured various additional extra-CNS anomalies. The association of complete vermal agenesis and trisomy 13 has not been previously reported. Our experience with this series suggests that supplementation with vaginal fetal sonography is a valuable tool for obtaining a more accurate view of the posterior fossa whenever a cyst or a cyst-like abnormality is detected by transabdominal sonography. A finding of isolated vermal agenesis appears to mandate a careful search for additional anomalies and the performance of karyotype analysis. PMID- 9358570 TI - Coelocentesis: a study of short-term safety. AB - Coelocentesis was performed in 20 singleton pregnancies at 6-10 weeks of gestation and 2-13 days before planned termination. The control group consisted of 100 women who were also undergoing planned termination and were matched with the study group for maternal age and gestation. During the follow-up period, there were five miscarriages (25 per cent) after coelocentesis and five in the control group (5 per cent) (chi 2 = 6.68; P < 0.01). The high risk of miscarriage following coelocentesis makes the technique unsuitable for early prenatal diagnosis. PMID- 9358571 TI - Fetal bradycardia following cordocentesis. AB - Several clinical investigations on the course and outcome of pregnancies following cordocentesis have mentioned the occurrence of fetal bradycardia at the time of umbilical cord puncture. The prognostic impact of this common complication has remained controversial. Our purpose was to investigate the prevalence and the short-term and long-term consequences of fetal bradycardia associated with cordocentesis. This study included all 339 cordocenteses performed in 290 fetuses at the Division of Prenatal Diagnosis and Therapy, University of Vienna, between 1991 and 1994. Clinically significant bradycardia was defined as a drop in the heart rate to less than 100 beats/min for a period of > or = 60 s. Bradycardia during or immediately after cordocentesis was observed in 13 cases (3.8 per cent). The fetal/neonatal loss rate per procedure was 61.5 per cent (8/13) in cases with bradycardia and 3.1 per cent (10/326) in those without bradycardia (P < 0.001). Early gestational age and hydrops fetalis correlated significantly with the development of bradycardia at cordocentesis. The other risk groups, including fetuses with intrauterine growth retardation, the puncture site, and the number of puncture attempts did not correlate with fetal bradycardia. Our results indicate that prolonged fetal bradycardia during or after cordocentesis is characteristic of a group of fetuses with an especially unfavourable prognosis. PMID- 9358572 TI - Clinical application of preimplantation diagnosis for myotonic dystrophy. AB - Myotonic dystrophy (DM) or Steinert's disease is a progressive autosomal dominant disease characterized by increasing muscle weakness, myotonia, cataracts, and endocrine abnormalities such as diabetes and testicular atrophy. The gene for DM was cloned in 1992 and the mutation was shown to be an expanded trinucleotide (CTG) repeat. A polymerase chain reaction (PCR)-based assay was described soon after that would allow (prenatal) diagnosis of the disease. Based on these PCR assays, we have developed a method for carrying out single-cell PCR for DM. In preimplantation diagnosis, embryos obtained in vitro are checked for the presence or absence of a disease, after which only embryos shown to be free of the disease under consideration are returned to the mother. A single-cell assay was developed for preimplantation diagnosis in couples where one of the parents is afflicted with DM. Twenty intracytoplasmic sperm injection (ICSI) cycles were carried out in eight patients and between one and four embryos were replaced in 17 out of 20 cycles. Two of the patients became pregnant and have had prenatal diagnosis which has confirmed that they are unaffected. PMID- 9358573 TI - Prenatal diagnosis of trisomy 9: cytogenetic, fish, and DNA studies. AB - A cytogenetic survey and follow-up studies were performed in eight cases of full, mosaic, and pseudomosaic trisomy 9 prenatally diagnosed among 36,213 prenatal samples in our department between August 1970 and July 1996. Besides conventional chromosome analysis, interphase fluorescent in situ hybridization (FISH) was employed. FISH turned out to be a rapid and accurate method for verification of trisomy cell lines and could provide additional information to the prenatal cytogenetic results. FISH also enables the study of uncultured specimens of amniotic fluid, not accessible for traditional cytogenetic analysis. In three cases, retrospective DNA analysis showed the supernumerary chromosome 9 to be of maternal origin. The disomic cell lines in both mosaic trisomy 9 cases showed maternal uniparental disomy. PMID- 9358574 TI - Biochemical markers of trisomy 21 and the pathophysiology of Down's syndrome pregnancies. AB - Using biochemical and immunocytochemical methods, we have investigated endogenous levels of various markers in tissues obtained from 67 Down's syndrome pregnancies after therapeutic abortion in the second trimester and in corresponding tissues from unaffected abortuses. Alpha-fetoprotein (AFP), intact and free beta human chorionic gonadotrophin (hCG), pregnancy-specific beta-1 glycoprotein (SP-1), placental alkaline phosphatase (PALP), pregnancy-associated plasma protein A (PAPP-A), and gamma glutamyl transferase (GGT) were investigated in placental tissue; AFP and GGT in fetal liver; and GGT in fetal intestine. The results indicate that maternal serum levels of placental products reflect those found in the placenta: intact hCG, free beta hCG, and SP-1 levels were elevated in Down's syndrome pregnancies, while PAPP-A and PALP levels were little changed. This suggests that membrane passage of these markers is not affected but there is altered synthesis of hCG and SP-1. AFP levels were strikingly elevated in placental homogenates and unchanged in liver homogenates from Down's syndrome pregnancies, while the levels in maternal serum were reduced, pointing to a possible transport defect specific to AFP. GGT levels were high in placenta and liver from Down's syndrome pregnancies but low in fetal intestine. PMID- 9358576 TI - The sonographic detection of early first-trimester conjoined twins. AB - The transvaginal transducer may be utilized to manipulate juxtaposed early first trimester twins in order to distinguish between conjoined and separate embryos. PMID- 9358577 TI - Mutation analysis for prenatal diagnosis of hereditary tyrosinaemia type 1. AB - Hereditary tyrosinaemia type 1 is a rare but serious metabolic disorder with an autosomal recessive mode of inheritance. We describe the prenatal diagnosis of an affected fetus performed by DNA-mutation analysis and a subsequent pregnancy with a healthy child in the same family. PMID- 9358575 TI - The effect of chorionic villus sampling on the number of fetal cells isolated from maternal blood and on maternal serum alpha-fetoprotein levels. AB - Fetal cells are present in the circulation of pregnant women and can be isolated using density gradient centrifugation and magnetic cell sorting. In the present study, maternal cell preparations were depleted for CD45- and CD14-positive cells and enriched for CD71-positive cells. The number of fetal nucleated cells was determined using fluorescence in situ hybridization for X and Y chromosomes. Analysis of maternal blood samples taken before and after transabdominal chorionic villus sampling (TA-CVS) showed an increase in the number of fetal cells in 10 out of 19 male pregnancies after the invasive procedure. This cellular transfusion was found to correlate with elevated maternal serum alpha fetoprotein levels. TA-CVS-induced cellular transfusion may form a good in vivo system to optimize fetal cell isolation procedures and to study fetal cell dynamics and characteristics. PMID- 9358578 TI - Prenatal diagnosis of a highly undifferentiated brain tumour--a case report and review of the literature. AB - Intracranial tumours, often presenting with progressive hydrocephalus, are rare congenital diseases accounting for 0.5-1.5 per cent of all cases of brain tumours diagnosed during childhood. The differential diagnosis includes vascular malformations, infarctions, and haemorrhages. Sonographic signs suggestive of glioblastoma, teratoma, and astrocytoma do not establish the histological diagnosis, however. We report a case of an undifferentiated fetal glioma detected at 29 weeks' gestation. The diagnosis of an undifferentiated brain tumour was suspected by sonography because of the lack of normal brain structures in conjunction with a diffuse echogenic central lesion and an external hydrocephalus. Because of the very poor prognosis, we induced labour by intravaginal and intravenous administration of prostaglandin E2 and achieved the vaginal delivery of a stillborn child whose head circumference corresponded to 38 weeks of pregnancy. Histological and immunochemical features of this undifferentiated congenital glioma (glioblastoma) are presented. PMID- 9358579 TI - Prenatal diagnosis of vein of Galen aneurysmal malformation: report of two cases with proposal for prognostic indices. AB - Vein of Galen aneurysmal malformation (VGAM) is a rare, intracranial vascular anomaly that, until recently, has usually been diagnosed postnatally. Today, however, with the advances in high-resolution ultrasonography and colour Doppler, prenatal diagnosis is relatively easy. Due to novel intravascular embolization techniques, the prognosis of neonates with VGAM has markedly improved. A healthy infant with normal neurodevelopmental and cardiovascular status is now a reality. For the best outcome, however, careful planning of the appropriate time, mode, and place of delivery should be undertaken. To achieve this goal, in utero prognostic factors should be determined. This report illustrates, for the first time, prenatal ultrasonographic indices that may predict the outcome in two cases with a prenatal diagnosis of VGAM. The indices included mapping of intracranial feeding arteries, assessment of the width of the straight sinus, assessment and measurement of flow in the straight sinus, existence of 'steal' retrograde aortic flow, and the appearance of high-output cardiac state. By using these prenatal ultrasonographic parameters, fetal outcome may be predicted and appropriate management decided upon. PMID- 9358580 TI - In utero ultrasonographic diagnosis of an aberrant umbilical vein associated with fetal hepatic hyperechogenicity. AB - Intra-hepatic abnormalities of the fetal umbilical venous system are poorly documented and clinically not well understood. A case of routine ultrasound examination at 23 weeks' gestation demonstrating foci of hepatic hyperechogenicity and cardiomegaly is presented. Colour Doppler detected absence of flow in the ductus venosus and markedly increased blood flow through an aberrant channel connecting the umbilical vein with the right atrium. The pregnancy was terminated and anomalous venous drainage of the umbilical vein into an enlarged hepatic vein was found, as well as hepatic congestion and focal hepatic necrosis and calcifications. Incidental findings of fetal hepatic hyperechogenicities require colour Doppler investigation of the intra- and extra hepatic venous systems. We propose that a thrombo-embolic mechanism may be involved in the pathogenesis of these lesions. PMID- 9358581 TI - Prenatal clinical expression of 3-methylglutaconic aciduria: Barth syndrome. PMID- 9358582 TI - Prenatal DNA diagnosis on demand--a possible new approach to DNA service provision. PMID- 9358583 TI - Current awareness in prenatal diagnosis. PMID- 9358584 TI - Prostate cancer brachytherapy. PMID- 9358585 TI - Ultrasound anatomy for prostate brachytherapy. AB - Even for surgeons experienced in prostate anatomy, the manipulation of perineally placed needles into the prostate under the guidance of an ultrasound probe in the rectum can be disorienting. For those embarking on ultrasound-guided brachytherapy for prostate cancer, there can be no substitute for experience gained performing transrectal ultrasound examinations. A review of the anatomy of the region is presented, together with images showing how that anatomy appears sonographically. Some of the concepts involved in the role of prostate planimetry for dosimetry planning will be touched upon. PMID- 9358586 TI - Brachytherapy in cancer of the prostate: an historical perspective. AB - Carcinoma of the prostate in the United States has increased dramatically in the last few years due to improved detection methods including prostatic specific antigen testing and transrectal ultrasound. More than half of all prostate cancers are discovered while still localized. Radical prostatectomy and definitive radiation are reserved for patients in good health, who have localized disease. Brachytherapy, with its inherent ability to deliver a high dose to an organ-confined tumor, while minimally irradiating the surrounding tissues, has successfully competed with external beam for the treatment of early prostatic tumors. Their respective role is constantly under scrutiny and re-evaluation to improve the accuracy of delivery of radiation. The present review focuses on the role of brachytherapy for treatment of early cancer of the prostate over the span of this century and its future in the next millennium. PMID- 9358587 TI - The Baylor College of Medicine experience with gold seed implantation. AB - Advances in imaging technology and implant technique have led to the resurgent interest and practice of brachytherapy for the treatment of prostate cancer. Brachytherapy is a form of radiation treatment in which radioactive sources are placed directly into the tumor; it offers the advantage of maximizing the radiation dose delivered to the tumor while sparing the adjacent normal tissue. Permanent implants have become an important component of radiation delivery. Interstitial gold radioisotope (Au-198) implants for prostate cancer were introduced at Baylor College of Medicine in 1965. The rationale for using Au-198, instead of the two most commonly used radioisotopes, Palladium-103 (Pd-103) and Iodine-125 (I-125), is discussed, and the Baylor implant technique is compared to that used in other centers. Retrospective review divides the patient population into pre-ultrasound versus post-ultrasound eras. Dosimetric calculation and disease control with the Au-198 seed implant for prostatic cancer are reviewed for the two different eras; toxicity is evaluated in the post-ultrasound era only. In the pre-ultrasound era, 510 patients were treated with pelvic lymph node sampling and gold seed insertion of the prostate followed by external beam radiation. In the post-ultrasound era, 54 patients were treated definitively with ultrasound-guided transperineal Au-198 implant followed by external beam irradiation. A small group of 30 patients in the post-ultrasound era were evaluated for the efficacy of Au-198 re-implantation for locally recurrent disease. PMID- 9358588 TI - Gold seed implantation in prostate brachytherapy. AB - Interstitial brachytherapy for treatment of prostate cancer with radioactive gold -initially with liquid gold and later with seed technique--is based on an experience of more than four decades. With biopsy results approaching a 80% negative rate, and, at 5 years, a cancer specific survival of 100% for Stages A and B1, 90% for Stage B2, and 76% for Stage C, this form of treatment offers an effective and well-tolerated alternative mode of therapy for patients with localized prostate cancer. PMID- 9358589 TI - The role of prostatic planimetry using transrectal sonography in prostatic diseases. AB - Transrectal sonography (TRS) remains the best method for prostatic planimetry, providing much information of value for the diagnosis and treatment of prostatic diseases, particularly of prostatic cancer. Currently, TRS is a routine tool in the detection and treatment of prostatic cancer. Prostatic planimetry is also available for a better understanding of elevated prostate-specific antigen (PSA). PSA indices modified by prostatic volume are applicable to the detection of prostatic cancer. The most critical application of prostatic planimetry is in monitoring prostatic cancer after treatment. Prostatic volume decreased exponentially after androgen deprivation therapy without exception in patients with prostatic cancer. The parameter "tau" (Tau), calculated from kinetic analysis based on sequential planimetric volumetry using TRS after the initiation of treatment, is a reliable prognosticator in an individual patient. Furthermore, tau predicts the manner of disease progression, and local or metastatic progression. Thus, transrectal ultrasonic prostatic planimetry is of special value in prostatic cancer. PMID- 9358590 TI - The history of interstitial brachytherapy of prostatic cancer. AB - The history of interstitial brachytherapy of the prostate began in 1917 when Barringer inserted radium needles transperineally into the prostate, guided by a finger in the rectum. In 1952, Flocks et al. injected radioactive gold solution in prostatic cancer during open operation. In 1972, Whitmore et al. described retropubic Iodine-125 seed implantation through an open operation with pelvic lymph node dissection. The basis for ultrasound (US)-guided seed implantation was established in the late 1960s by Kratochwil's description of the first puncture transducer for one-dimensional US-guidance and Watanabe's pioneer work in prostatic screening. Our group performed the first punctures guided by static two dimensional ultrasonic scanning (1969) and by dynamic ultrasound scanning (1974). During the 1970s, US-guided biopsies of almost all abdominal organs were performed and, in 1981, we introduced US-guided seed implantation of abdominal tumors and developed a technique for precise needle placement in the prostate guided by transrectal ultrasound. This was followed in 1983 by a technique for seed implantation guided by transrectal (transverse) ultrasonic scanning. That same year, Fornage described prostatic biopsy guided by transrectal longitudinal scanning. In 1990, we developed the seed implantation technique further by combining transverse and longitudinal scannings. In the late 1980s, Ragde from Seattle transferred the technique to the U.S., and since then, Ragde, Blasko, and others have treated numerous patients, refined the technique, taught many courses, and published extensively. They have obtained very promising prostate specific antigen (PSA)-based results which compare favorably with radical prostatectomy and external beam radiation in the treatment of prostatic cancer. PMID- 9358591 TI - Brachytherapy for clinically localized prostate cancer: results at 7- and 8-year follow-up. AB - In recent years, there has been a resurgence of interest in interstitial radiation as a cost-effective and efficient method of treating organ-confined prostate cancer. We describe our 7- and 8-year results with transperineal Iodine 125 and Palladium-103 implantation. A total of 551 consecutive patients were treated. Of these, 320/551 (58%) received implant alone (Group I), and 231/551 (42%)--considered higher risk patients--were also treated with a modest dose (45 Gy) of external beam irradiation (Group II). The median follow-up for Group I was 55 months, and for Group II, 60 months. At 7 years, the actuarial freedom from biochemical failure (prostate-specific antigen (PSA) < or = 1.0 ng/mL) was 80% in Group I patients, and, at 8 years, 65% in Group II patients. Morbidity was minimal if patients had not undergone prior transurethral prostate resections. The results indicate that interstitial radiation is a valid treatment for clinically localized prostate cancer. PMID- 9358592 TI - Current issues in techniques of prostate brachytherapy. AB - Adenocarcinoma of the prostate is the most common malignancy diagnosed among men in the United States today. Brachytherapy permits conformal radiotherapy and dose escalation, and it offers the convenience of a single-day outpatient procedure which is very attractive to patients with a busy life-style. The reported potency preservation rates with brachytherapy are superior to both external beam radiation therapy (EBRT) and surgery. The older retropubic techniques have been replaced by ultrasound or CT-guided transperineal techniques. Prostate brachytherapy may be temporary or permanent, and the planning techniques for either approach are similar. This review briefly discusses the advantages and limitations of each. Temporary techniques may be used with low dose rate or high dose rate applications. The basic steps include assessing prostate volume by any diagnostic modality (CT or ultrasonography), determining total activity needed to encompass the gland and deliver the appropriate minimum peripheral dose, and determining the pattern of placement of the seeds within the gland. Preplanning may be done either by ultrasound or by CT. The operative technique requires the visualization of the prostate in three dimensions and is performed using combination of ultrasound and fluoroscopy or fluoroscopy in two axes. The New York Hospital technique employs CT-based preplanning along with ultrasound and fluoroscopy during the operative procedure. Special circumstances that necessitate neoadjuvant hormonal therapy include interference from the pubic arch and large volume glands. An analysis of patients with stage T2a disease treated at the New York Hospital-Queens, from 1990-1995, reveals an actuarial clinical freedom from relapse of 79% at 5 years and a 5-year biochemical freedom from relapse of 64% which is comparable to that reported for similar risk groups of disease by other centers. Potency is preserved in greater than 80% of patients in our series. Patient selection criteria include the pre-treatment prostate specific antigen (PSA) level, tumor grade (Gleason), stage of disease, and presence or absence of bilateral positive biopsies and/or perineural invasion. Based on our review of the literature and our clinical results, we have divided patients with prostate cancer into good, intermediate and poor risk groups. We recommend brachytherapy as the sole procedure for good risk patients, and a combination of external beam radiation therapy (EBRT) and brachytherapy for the intermediate risk group. Future avenues for research include a search for improved imaging techniques and possibly newer isotopes. PMID- 9358594 TI - Quality of life after permanent prostate implant. AB - Quality of life is one of several endpoints commonly studied in prostate cancer treatment. It refers to how well an individual is functioning in life and his total sense of well being. There is increasing recognition that cancer therapy impacts significantly on the patient's ability to pursue relational, occupational and social interests. Fifty-one patients with clinically localized prostate cancer who had undergone transperineal permanent prostate implantation were evaluated. All patients were clinically staged as T1c or T2a and received an implant alone with Iodine 125 or Palladium 103 as definitive treatment. Six months after implant, data was collected using the European Organization for Research and Treatment of Cancer (EORTC) genitourinary group questionnaire and supplemental questions. Urinary symptoms such as nocturia, hesitancy, frequency, and dysuria were the most pronounced in the first few months after the implant and then decreased in most of patients; 40% noticed that they urinated more frequently and 17% had mild dysuria. All patients denied hematuria and none reported incontinence. Few patients reported any psychological distress or disruption in social or family life; none reported disruption in economic status or viability. All fifty-one patients said that they would have an implant again as definitive treatment. Seventy-nine percent reported an excellent quality of life post-implant. While survival is clearly a central goal of treatment for prostate cancer, the nature of this malignancy compels clinical attention to the quality of the patient's life after treatment. Sexual quality and function are maintained in the majority of patients and there is minimal interruption of their social and economic function. PMID- 9358593 TI - The effect of prognostic factors on therapeutic outcome following transperineal prostate brachytherapy. AB - The objectives of this study were to examine the effect of both disease and treatment related prognostic factors on biochemical control and post-treatment biopsy. Two-hundred fifty-eight patients underwent interactive ultrasound guided transperineal prostate implantation for T1-T2 prostate cancer using Iodine-125 (139 patients) and Palladium-103 (119 patients) and were followed from 6-67 months (median, 19). Hormonal therapy with 3 months of leuprolide and flutamide prior to implantation and two months given after the implant was used in 96 patients. Pre-treatment prostate-specific antigen (PSA) had the most significant effect on biochemical failure. Freedom from biochemical failure (FFBF) rates at 4 years were 75% for patients with PSA 1.3-10 ng/ml (144), 74% for patients with PSA 10.1-20 ng/ml (73), and 34% for patients with PSA > 20 ng/ml (41) (P = 0.0004). Gleason score also had a significant effect on FFBF rates. Four-year rates were 81%, 65% and 47% for patients with scores of 2-4 (68), 5-6 (130), and > or = 7 respectively (60) (P = 0.01). These two factors were also significant in multivariate analysis (P = 0.002, 0.007, respectively). Gleason score was the only factor to significantly affect post-treatment biopsy results. Patients with scores of 2-6 had 85% (63/ 74) negative 2-year biopsies versus 62% (13/21) for patients with scores > or = 7 (P = 0.02). Low-risk patients (PSA < or = 10 ng/ml, scores < 7 and stage < T2a) had a 4-year FFBF rate of 88% as compared to 60% for high-risk patients (PSA > 10 ng/ml, score > 6 or stage > or = T2b) (P = 0.02) and had a 95% negative biopsy rate versus 76% for high-risk patients (P = 0.06). Low risk patients demonstrate high FFBF and negative biopsy rates following implantation. Patients presenting with higher risk prognostic factors such as PSA > 20 ng/ml or Gleason scores > or = 7 may require more aggressive treatment regimens. PMID- 9358595 TI - Social work in mental health: trends and issues. PMID- 9358596 TI - Recovery and empowerment for people with psychiatric disabilities. AB - In this paper the concept of recovery from major mental illness and the empowerment process are explored. Subjective experiences from the author's own journey of recovery from mental illness as well as others are explored. The concept of recovery as a journey, not a destination or "cure" is emphasized. It is noted that one must recover not only from mental illness, but also from internalized stigma, low expectations and dehumanizing clinical practices. Suggestions for the clinical practitioner who wishes to support the recovery and empowerment process are also given. PMID- 9358597 TI - Community care and the origins of psychiatric social work. AB - Psychiatric social work began in the early twentieth century as a result of a movement for community care of the mentally ill. Formerly preoccupied with institutional care of the mentally ill, psychiatrists now became interested in the control and prevention of mental illness in the community. This paper reviews the contemporary literature on early psychiatric social work. The paper then discusses the mental hygiene movement and early psychiatric social work practice. It concludes with a consideration of the changes in psychiatric social work which accompanied World War I. PMID- 9358598 TI - Strategies for helping young adults with severe mental disorders. AB - Severe mental illness is defined in terms of psychosocial development and attention is given to socio-economic forces in these patients' lives. The difficulties faced by the young, severely mentally ill persons in the United States is discussed and data regarding prevalence and treatment strategies are outlined with implications for individual and family therapy modalities. Programs with a wide range of psychosocial, vocational and housing support systems together with counseling are emphasized as integral components if young, emotionally disabled persons are to be sustained in the community. The lay and professional communities must be mobilized to provide the needed resources. Available and accessible innovative programmatic concepts together with a more hopeful attitude on the part of professionals can make a difference. PMID- 9358599 TI - Social networks and psychological disability among housed and homeless users of self-help agencies. AB - We look at the effects of psychological disability on social networks and support of homeless and non-homeless individuals. We analyze a survey of 310 long-term users of client-run mental health agencies. Psychological disability is negatively associated with network characteristics for housed individuals, but not for the homeless. There is a positive relationship between psychological distress and network size for the homeless who receive SSI while homeless individuals who do not receive SSI show a negative, non-significant association. We suggest the financial resources of SSI enable network members to become expressively involved with homeless individuals with relatively more psychological disturbance. PMID- 9358600 TI - Discharge planning and community housing in Ontario. AB - As hospital budgets in Ontario (and elsewhere) continue to shrink in the face of governmental fiscal pressure, bed closures lead to the discharge of increasingly vulnerable persons. Many of these persons have no family and no obvious place to go. Community supports to assist people outside the hospitals are not provided at a level commensurate with the need. The result is inadequate housing, social isolation, non-existent care and, in too many cases, reinstitutionalization and/or preventable deaths. This paper describes the process by which vulnerable adults wind up in unsuitable community settings, as a result of ill-conceived deinstitutionalization in the province of Ontario. It places a particular focus on the difficult role played by the discharge planner as conduit from hospital to community. The planner is often caught in the middle, facing hospital (and physician) directives to empty beds precipitously, alongside an acute shortage of suitable housing in the community. Departing patients are often sent to settings that lack any form of governmental inspection, regulation, licensure, or control: they are at the mercy of often indifferent and, at times, overtly rapacious landlords who may take the welfare cheque and give little in return. Selected case material, including one recent inquest, highlight the difficulties. PMID- 9358601 TI - Working collaboratively with families. AB - Research studies indicate that significant tension characterizes the relationships between providers and families whose relative is being treated in the mental health system. The author recommends that genuinely collaborative relationships be developed in order that people receiving treatment receive optimal care. Collaboration is defined, barriers identified, and ways to overcome these barriers suggested. PMID- 9358602 TI - The universality of a self-help program of American origin: narcotics anonymous in Israel. AB - A phenomenological field study of Narcotics Anonymous (NA) in Israel focused on the way a self-help program, based on American Christian ideology was adopted in Israel. Acculturation problems were anticipated, due to cultural, demographic and religious differences. Participant observations and open-ended interviews supplied the raw data. Emphasis was placed on the factors and processes definable as typically American: voluntarism and pragmatism, personal sharing as a basis for relationships, spiritual rather than religious faith, the idea of a "personal God" guiding individuals, faith in God without religious tradition and formal ritualism. The results showed that, for the substance-dependents, the issue is generally irrelevant, and they accepted most "American" components of the program unquestioningly. However, two discrete features of Christian ideology required conscious incorporation by NA's Israeli members: (1) the concept of a "Loving God" who is non-punitive, which for many members was opposed to their traditions and upbringing, and (2) kneeling to pray, a recommendation which many members initially found problematic. The conclusion denotes a factor facilitating the transfer of therapeutic programs from one culture to another--that of personal suffering as a universal domain. It transcends all cultural boundaries and generates willingness to accept foreign concepts which reveal suffering and propose a pragmatic way to end it. PMID- 9358603 TI - The ethics of informed consent: a critical variable in the self-determination of health and mental health clients. AB - Informed consent is a critical variable in the self-determination of consumers receiving health and mental health services. Consent has become a more stringent requirement in recent years. However, providers emphasize legal requirements rather than the true participation of consumers--the "spirit" of informed consent. This article discusses the elements of informed consent, the issues that reflect the "spirit" of consent, and recommendations for actions that foster self determination in practice. PMID- 9358604 TI - Revamping mental health care in Israel: from the Netanyahu Commission to National Health Insurance Law. AB - This paper describes the basis for the reform in mental health care system in Israel as presented in the report of the Netanyahu Commission (State Commission for Investigation of Functioning and Efficiency of the Health Care System in Israel, 1990) and the report of the State Comptroller's office (State Comptroller, 1991). These reports pointed to seven major problem areas in the mental health care system: (1) segregation of mental health and general health care systems, (2) variations in availability of services across the country, (3) conflict of interests within Ministry of Health which provides services and oversight, (4) overuse of hospital based care and under use of community based care, (5) reliance on hospitals for custodial care, (6) lack of appreciation of mental health service needs of non-severely mentally ill, and (7) lack of regional service planning. The article describes these problems and the proposed solutions. PMID- 9358606 TI - Cholesterol biosynthesis inhibited by BM15.766 induces holoprosencephaly in the rat. AB - To confirm that blocking 7-dehydrocholesterol delta 7 reductase (7DHC reductase), as observed in Smith-Lemli-Opitz syndrome (SLOS), induces craniofacial defects, we tested BM15.766, which blocks 7DHC reductase but is chemically unrelated to the holoprosencephaly-inducing teratogen AY9944. Rats were given BM15.766 either in methylcellulose from days (D) 1 through D11 (3 treated groups: protocol A) or in olive oil from D4 through D7 (300 mg/kg/d: protocol B). The sera were sampled on D0, D3, and D5 or D6, D10, D14, and D21 to measure cholesterol and dehydrocholesterols in all groups and steroid hormones in protocol B. D21 fetuses showed the holoprosencephaly spectrum of malformations and the treated dams low cholesterol and accumulation of 7DHC, 8DHC, and trienols, as in SLOS-affected children. In the 3 dosage groups the malformations were dose-related and enzymatic cholesterol decreased to a plateau. The DHC reached 25-44% of the total sterols in the dams. In protocol B, one-third of the BM15.766-treated fetuses presented facial malformations and almost two-thirds pituitary agenesis. On D10, cholesterol reached a minimum and the DHC a maximum while estradiol 17 beta and progesterone were lowered, the latter decreasing in correlation with cholesterolemia. A sterol profile similar to that previously observed after AY9944 associated with a similarly high incidence of pituitary agenesis confirmed that time-limited inhibition of 7DHC reductase induces holoprosencephaly and that pituitary agenesis is the minor form of holoprosencephaly. PMID- 9358605 TI - Genetic landmarks for defects in mouse neural tube closure. AB - Many mutations cause neural tube closure defects (NTDs, exencephaly or spina bifida) in mice and the gene loci are widely distributed in the mouse genome. This compilation summarizes the map position of 40 mouse NTD mutations and the corresponding human linkage homology of each. It includes the nature of the gene product where known, and whether the NTD is part of a syndrome involving other developmental systems. Also listed are the several mouse strains known to have genetic susceptibility to exencephaly, with multifactorial genetic cause in at least one case. The purposes of this mouse NTD compilation are to enable recognition of patterns in genetic causes of NTDs, of molecular pathways essential for closure of specific regions of the mammalian neural tube, and of candidate regions for mapping loci contributing to human multifactorial NTDs. PMID- 9358607 TI - Fluorometric analysis of endocytosis and lysosomal proteolysis in the rat visceral yolk sac during whole embryo culture. AB - Using spectrofluorimetry and fluorescence microscopy, we analyzed the uptake and degradation of fluorescein isothiocyanate-conjugated bovine serum albumin (FITC albumin) by the rat visceral yolk sac (VYS) during whole embryo culture. Rat conceptuses exposed continuously to FITC-albumin had linear increases of both acid-soluble and acid-insoluble FITC fluorescence in the VYS. Smaller amounts of FITC fluorescence that were nearly all acid soluble accumulated in the extraembryonic fluid, while the embryo proper did not accumulate a significant amount of fluorescence. During a chase period following a pulse exposure to FITC albumin, FITC fluorescence in the VYS decreased linearly, while that in the extraembryonic fluid and culture medium increased. Addition of proteinase inhibitors to the culture medium together with FITC-albumin increased acid insoluble FITC-fluorescence in the VYS tissue but decreased acid-soluble fluorescent degradation products in the yolk sac, extraembryonic fluid, and the culture medium. Fluorescence microscopy of yolk sacs exposed to FITC-albumin revealed that the fluorescence was localized in apical vacuoles of the yolk sac epithelium and decreased substantially during a chase period. In conceptuses exposed to proteinase inhibitors, the yolk sac epithelium had enlarged vacuoles containing FITC-fluorescence whose clearance in pulse-chase experiments was effectively blocked. Overall, these data suggest that FITC-albumin resembles 125l albumin in its processing by the VYS and that the fluorescent protein is an attractive alternative tracer molecule for studies of the effects of embryotoxicants on yolk sac function during whole embryo culture. PMID- 9358608 TI - Induction of thermotolerance in early postimplantation rat embryos is associated with increased resistance to hyperthermia-induced apoptosis. AB - Previously we reported that hyperthermia (43 degrees C) induces cell death in neurulation stage rat embryos as part of the pathogenesis culminating in abnormal growth and development. We now show that hyperthermia-induced cell death occurs by a process termed apoptosis. DNA fragmentation, a hallmark of apoptosis, was noted as early as 2.5 hr after embryos were exposed to 43 degrees C. A smaller but significant increase in DNA fragmentation was also observed in embryos exposed to 42 degrees C, but only at the 5 hr time point. In control embryos, TUNEL-positive apoptotic bodies were consistently observed in the neuroepithelium at the point of neural tube closure and in the optic stalk. In embryos exposed to 43 degrees C, the number of TUNEL-positive apoptotic bodies was significantly increased. Using both gel electrophoresis and TUNEL, we also show that the induction of thermotolerance is associated with a significant reduction in DNA fragmentation. Together our results show that specific programmed cell death and hyperthermia-induced cell death correlate with internucleosomal DNA fragmentation characteristic of apoptosis. Finally, we show that the induction of thermotolerance in rat embryos is associated with a significant reduction in internucleosomal DNA fragmentation and associated apoptosis. PMID- 9358609 TI - Neonatal treatment with tamoxifen causes immediate alterations of the sexually dimorphic nucleus of the preoptic area and medial preoptic area in male rats. AB - Tamoxifen is an antiestrogen widely used for the treatment of breast cancer. Current evolutions in preventive strategies to include healthy premenopausal women warrant the study of its developmental toxicity. Perinatal treatment of male rodents with tamoxifen caused reproductive tract lesions and sexual behavior deficits similar to those induced by diethylstilbestrol (DES). Those abnormalities could originate, at least in part, from lesions of the hypothalamic pituitary axis. The initial alterations caused by tamoxifen in the hypothalamic medial preoptic area (MPOA) and sexually dimorphic nucleus of the preoptic area (SDN-POA) were studied in 6-day-old male rat pups treated with 100 micrograms tamoxifen (group 1), 1 microgram DES (group 2) or vehicle (group 3) from PN1 to 5. In situ hybridization was performed to analyze the expression of the GAP-43 gene, a marker of neuronal differentiation, and morphometry was used to study the neuronal density in the SDN-POA and MPOA and the volume and number of neurons in the SDN-POA. Tamoxifen reduced severely the volume and neuron numbers in the SDN POA (46.1% and 47.8% of controls, respectively). The neuronal density of the MPOA was not modified. GAP-43 gene expression was decreased as demonstrated by a greater percentage of unlabeled neurons (grade 0) mirrored by a lesser percentage of intensely labeled ones (grade 2) in the SDN-POA and MPOA. In contrast to the effects of the antiestrogen, DES did not affect the above endpoints. These data indicated that developmental exposure of male rat pups to tamoxifen-induced immediate neuronal loss in one and altered differentiation in two hypothalamic areas crucial to reproduction. How those initial alterations contribute to the pathogenesis of the reproductive disorders observed in the adult male needs further investigation. PMID- 9358610 TI - Ex-vivo whole blood cultures for predicting cytokine-release syndrome: dependence on target antigen and antibody isotype. AB - Ex-vivo whole blood assays have been evaluated for their ability to accurately predict the risk of a first-dose cytokine reaction developing in vivo following therapeutic antibody infusion. Tumour necrosis factor alpha (TNF alpha) release was rapidly detected in cultures incubated with either anti-CD52 antibodies of the human IgG1 or rat IgG2b isotype, and to a lesser extent with a human IgG4 isotype. Endotoxin contamination of the antibodies was not responsive for cytokine release, since polymixin B failed to inhibit cytokine release using concentrations of this antibiotic which neutralized the enhanced cytokine release seen from LPS-spiked antibody. A rat IgG2b antibody to CD45 and a human IgG1 anti CD3 also induced significant TNF release, however, an aglycosyl anti-CD3 mutant devoid of adverse side-effects in vivo, did not result in cytokine release in vitro. Since the pattern of cytokine release seen following the clinical use of these antibodies was in good agreement with the findings of the ex-vivo whole cultures, this demonstrates the usefulness of this assay to predict cytokine release in vivo. PMID- 9358611 TI - The development of anti-CD79 monoclonal antibodies for treatment of B-cell neoplastic disease. AB - The B-cell antigen receptor (BCR) consists of cell surface IgM associated with the CD79 alpha/beta heterodimer. In this paper we describe a panel of monoclonal antibodies (mAbs) recognising the extracellular regions of human CD79 alpha and beta. FACS analysis demonstrated that the mAbs bind to a range of Burkitt's lymphoma lines, a mouse B-cell line (JO-72) transfected with human CD79 alpha and beta, and tumour biopsies from NHL patients. The specificity of the mAbs was confirmed by immunoprecipitation. The Ka for the binding of IgG from the anti CD79 alpha mAbs to cell surface CD79 alpha on Ramos cells was 3 x 10(8) M-1, and their maximum level of binding, 1.7-2 x 10(5) molecules/cell, matched that obtained with anti-Fc mu and anti-Fd mu mAbs. All four anti-CD79 beta mAbs were of lower affinity. Interestingly, in growth arrest studies, we found that while all anti-Fc mu mAbs caused profound inhibition of proliferation of Ramos cells, a range of other anti-BCR mAbs, which included the anti-CD79, anti-Fab mu, anti gamma and anti-idiotype reagents, all performed poorly giving a maximum of 25% inhibition. These differences in performance are believed to relate to the ability of anti-BCR mAbs to cross-link neighbouring surface BCR and suggest that, unlike anti-Fc mu which favours cross-linking, most of these mAbs are binding in a monogamous, non-cross-linking, union with the BCR. PMID- 9358612 TI - Potential use of in vitro anterior chamber-associated immune deviation (ACAID) for the immunotherapeutic prevention of autoimmune disease and graft rejection. PMID- 9358613 TI - Rational development of tumour antigen-specific immunization in melanoma. AB - The identification and molecular characterization of antigens expressed on tumour cells, but not on most normal host tissues, has opened the possibility of specific immunization in the therapy of cancer, particularly of melanoma. Most antigens defined are class I MHC-binding peptides recognized by CD8+ cytolytic T lymphocytes (CTL). Methodologies for active immunization to induce effective anti tumour CTL are under development in a number of laboratories, and some of these approaches have entered clinical trials. Optimization of the anti-tumour immune response will depend on a thorough knowledge of the signals required for T cell activation, differentiation, and inactivation. PMID- 9358614 TI - Peripheral blood stem cell transplantation. AB - High dose chemo/radiotherapy requiring autologous haemopoietic stem cell support is increasingly used in a variety of malignant disorders. Mobilised peripheral blood stem cells (PBSC) have largely replaced the use of autologous bone marrow due to more rapid haemopoietic reconstitution with less resource use including blood and platelet transfusion requirements. PBSC graft adequacy is monitored by CD34+ cell and granulocyte-monocyte-colony-forming-cell measurements, and thresholds for rapid engraftment have been determined. Studies are in progress to determine the optimal mobilisation regimens that will permit the achievement of the necessary progenitor thresholds with only one or two aphereses. This will facilitate the use of multiple cycles of high dose therapy and possibly the use of PBSC collected by venesection rather than apheresis. PBSC are also increasingly used in the allogeneic setting where specific mobilisation protocols not using cytotoxic drugs are employed. These technical advances will aid the execution of large trials to determine the efficacy of high dose therapy. PMID- 9358615 TI - Evaluation of a malaria antibody ELISA and its value in reducing potential wastage of red cell donations from blood donors exposed to malaria, with a note on a case of transfusion-transmitted malaria. AB - BACKGROUND AND OBJECTIVES: Blood donations are often wasted for lack of a satisfactory procedure to evaluate donors potentially exposed to malaria. MATERIALS AND METHODS: We evaluated a commercial ELISA for the detection of antibodies to malaria and compared it with an immunofluorescent antibody test (IFAT). RESULTS: When 5,311 sera from routine non-exposed donors were tested, 24 (0.45%) were positive by the ELISA, using a Plasmodium falciparum antigen. Seventeen were subjected to confirmatory testing but none were positive by IFAT. Of 1,000 donors potentially exposed in endemic areas 15 (1.5%) were repeatably reactive by ELISA. 10 of these were tested by IFAT and 2 were positive. When 150 patients attending the Hospital for Tropical Diseases in London with acute malaria were tested, 73% of those infected with P. falciparum were repeatably reactive for malarial antibodies by ELISA and 56% with Plasmodium vivax. Of 88 stored clinical sera tested by both IFAT and ELISA 56 were positive by IFAT and of these 52 (93 degrees/0) were positive by ELISA. CONCLUSION: The ELISA is sufficiently sensitive and specific to screen at-risk donors. Its use could safely retrieve 40,000 red cell units currently discarded each year in Great Britain. PMID- 9358616 TI - Virological and immunological data of AIDS patients treated by passive immunotherapy (transfusions of plasma rich in HIV-1 antibodies). AB - BACKGROUND AND OBJECTIVES: In human immunodeficiency virus (HIV) infections, passive immunotherapy can be carried out through infusions of virus-inactivated plasma from symptomless HIV-infected persons with abundant HIV antibodies. MATERIALS AND METHODS: We carried out a prospective, randomized, double-blind, controlled, passive immunotherapy study, which compared two groups. One received plasma rich in HIV antibodies, the other a standard seronegative plasma. RESULTS: Measurement of the plasma HIV RNA load showed in both groups a significant decrease in the mean viral copy number at the end of the first month, followed by an increase at the third month. Beyond the third months, a significant decrease in viral load was observed only in the treatment group. A significant difference in favor of the treatment group was observed for plasma viremia by HIV culture. For the cytokines involved in the viral replication and for the immune activation markers such as neopterin and beta 2-microglobulin, the biological analysis in plasma failed to show a significant difference in either group. Clinically, the treatment group benefited by delay in the appearance of the first AIDS-defining event and reduction in the cumulative incidence of such events. CONCLUSION: One possible interpretation is that passive immunotherapy affects plasma viral load, but there is no evidence that HIV-specific antibodies are exclusively responsible for the observed effects. PMID- 9358617 TI - Factor VII and activated-factor-VII content of prothrombin complex concentrates. The PCC Study Group. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to determine the potencies of factor VII (FVII) and of activated FVII (FVIIa) in prothrombin complex concentrates (PCC). MATERIALS AND METHODS: We examined 56 lots of PCC from 5 manufacturers. Three brands were licensed preparations, and 1 product series had been involved in thromboembolic complications. FVII and FVIIa were measured using a two-stage amidolytic assay and a specific clotting assay, respectively. We also quantified FVII clotting activity by a one-stage assay reflecting a mixture of FVII zymogen and FVIIa. RESULTS: All PCC contained substantial amounts of FVII, and FVIIa could be detected in all lots. There were marked differences between manufacturers and some significant variabilities between batches. The two lots involved in thromboembolic events contained considerably more FVIIa than the PCC still licensed. The lowest FVIIa potencies were observed in an experimental product series, indicating that PCC can be produced without activation of FVII during the manufacturing process. CONCLUSION: FVIIa is present in all PCC containing FVII. High FVIIa potencies may contribute to the thrombogenic potential of these preparations, and determination of FVIIa potencies should be included in the in vitro characterization of PCC. PMID- 9358618 TI - Use of SERVQUAL to assess clinicians' satisfaction with the blood transfusion service. AB - BACKGROUND AND OBJECTIVES: Limited information is available on the level of satisfaction of clinicians with services delivered by blood banks. The purpose of this study was to evaluate the satisfaction of clinicians with our blood transfusion service. MATERIALS AND METHODS: We prepared a questionnaire based on SERVQUAL, a method used to measure customers' appreciation of quality of service, by assessing the gap between perceived and expected quality. The questionnaire consisted of 14 items grouped according to five dimensions of quality of service: assurance, empathy, responsiveness, reliability, tangibles. Clinicians were asked to give two scores on a scale from 1 to 7 for each item, score (e) representing what they expected from an 'excellent' service, score (r) how they graded the service received. We considered wide differences in scores of service expectation and receipt for a question to be indicative of either service above expected levels (r > e) or service below expectation (r < e); similar scores for both expected and received service (within 1 point on the grading scale) were taken to indicate that the service received was that which was expected. RESULTS: A total of 184 questionnaires (49%) were returned. For the 14 items considered, the proportion of clinicians expressing levels of satisfaction similar to or above expectation ranged from 67 to 96%. Three critical areas, which clinicians considered important (expectation scores 6-7) were associated with satisfaction below expectation in more than 20% of responders. They were: clarity of procedures, clarity of blood request forms, and convenience of blood request and issuing times, which were rated as important by 77, 80 and 72% of clinicians, respectively. CONCLUSION: SERVQUAL was useful to gather information on the level of clinicians' satisfaction with our transfusion service. PMID- 9358619 TI - Transfusion-related immunomodulation in Chinese children with thalassaemia. AB - BACKGROUND AND OBJECTIVES: Multiple transfusions can induce immunomodulation. This study was carried out to investigate the immunological status of 50 transfusion-dependent children with beta-thalassaemia, taking into account that lymphocyte characteristics are affected by sex, age and race. We paid particular attention to the influence of transfusion and serum ferritin on the lymphocyte subsets which may be affected by the exposure to foreign antigens. MATERIALS AND METHODS: By multicolour immunofluorescent analysis using flow cytometry, we determined lymphocyte characteristics with regard to major subsets (T lymphocytes, B lymphocytes and NK cells), activation (membrane IL-2 receptor CD25, HLA-D) and memory/naive T cells (CD45RO/CD45RA). Data from 51 age- and sex balanced children served as controls. RESULTS: The normal Chinese children had higher NK levels than the beta-thalassaemia children. The levels of CD25 and HLA D indicated a broad-based increase in activation status. Memory T cells were also increased when compared with their normal counterparts. We found additional and more marked alterations in the lymphocyte subsets of those who had received over 100 transfusions. While levels of NK cells were inversely correlated with the number of transfusions, CD25+ cells increased with transfusions. CONCLUSION: Many multitransfused beta-thalassaemia children have altered levels of lymphocyte subsets compared with normals. What remains to be investigated is the long-term consequence of possessing low NK and non-MHC-restricted T cells (CD3+CD56+CD16+) and a high activation status in terms of resistance of infections and development of malignancy. PMID- 9358620 TI - Immunochemical characterization of the Rh CW antigen using human monoclonal antibodies. AB - BACKGROUND AND OBJECTIVES: Human monoclonal antibodies have been produced against various Rh antigens. We report here the production of another one against Rh C and the first such antibody against Rh CW. MATERIALS AND METHODS: Two heterohybridomas secreting IgG human monoclonal antibodies against Rh CW and Rh C (MS-353 and MS-242 respectively) were produced from alloimmunized donors. Their specificity was confirmed by serological testing. RESULTS: MS-353 reacted with all CW-positive phenotypes but not with any CW-negative phenotypes, whereas MS 242 reacted with all CW-positive and C-positive phenotypes. Using 125I-labelled antibodies, the average number of CW sites/red cell was 32,000 (R1WR1W, 15,200 (R1WR1), 19,800 (R1WR2), 15,300 (R1Wr), 26,200 (r'Wr), and the average number of C sites per red cell on equivalent CW/C-positive phenotypes was similar: 22,400 (R1R1), 21,800 (R1WR1), 12,300 (R1R2), 11,700 (R1WR2), 13,400 (R1r), 15,100 (R1Wr), 21,100 (r'r), 18,700 (r'Wr). MS-353 and MS-242 were mutually inhibitory. Both antibodies specifically immunoprecipitated a 33- to 34-kD polypeptide from 125I-surface-labelled cells. CONCLUSION: MS-353 is suitable for the serological detection of CW-positive cells, and the immunochemical data are entirely consistent with the report that the CW antigen is associated with a point mutation in the RHCE gene [Blood, 1995;86:1196-1201]. PMID- 9358621 TI - Gene frequencies of the HPA-1 and HPA-2 platelet antigen alleles among the Amerindians. AB - BACKGROUND AND OBJECTIVES: Platelet-specific alloantigens are important in neonatal alloimmune thrombocytopenia, posttransfusion purpura, refractoriness to platelet transfusions, and population genetics. Data are scarce on allele frequencies in ethnic groups other than whites and Asians. MATERIALS AND METHODS: Using allele-specific restriction enzyme analysis, we studied the distribution of HPA-1 and HPA-2 alleles in six Brazilian Amazon tribes of Amerindians, belonging to five different language stocks. We compared these with the values obtained for blacks and whites. RESULTS: Only the HPA-1a allele was found among 132 Amerindian chromosomes, compared with a gene frequency of HPA-1b of 0.115 and 0.113, respectively, among blacks and whites. The frequency of HPA-2b among the Amerindians (0.042) is lower than that obtained for blacks and whites (0.148 and 0.100, respectively), and the lowest thus far observed in a population of Asian origin. CONCLUSION: Differences in DNA polymorphisms in Amerindian populations have not only anthropological and genetic interest, but also practical applications when they involve coding regions that may change the functional or immunologic features of the protein. PMID- 9358622 TI - Genotyping of the human platelet antigen-1 by ELISA detection of allele-specific amplicons. AB - BACKGROUND AND OBJECTIVES: The purpose of the study was to establish a valid and efficient method for genotyping of the human platelet alloantigen-1 (HPA-1) in large sample numbers. MATERIALS AND METHODS: Digoxigenin- and fluorescein labelled allele-specific primers and a biotinylated common primer were included in the hot-start PCR. Amplicons were bound to avidin-coated plates to identify the products by ELISA. RESULTS: This approach reduced the number of PCR analyses for HPA-1 typing by half. PCR products were detected with high sensitivity and good reproducibility (interassay-CV: < 8%) in the ELISA. The typing of 100 blood donors with both PCR plus gel electrophoresis and ELISA-PCR showed identical results. CONCLUSION: This method offers efficient HPA-1 genotyping in large numbers of donors and patients. PMID- 9358623 TI - Hepatitis A transmission by factor IX concentrates: control by severe dry heat treatment at 80 degrees C. PMID- 9358624 TI - Epidemiology of GBV-C in Japan. PMID- 9358626 TI - A new procedure for the specific high-performance liquid chromatographic determination of hydroxyproline. AB - A procedure suitable for a selective high-performance liquid chromatographic (HPLC) analysis of the imino acid hydroxyproline in the presence of a large excess of amino acids is proposed. To deaminate the amino acids, the well-known reaction with nitrous acid is exploited. The N-nitroso derivatives of imino acids obtained are extracted in ethyl acetate, denitrosated by hydrobromic acid, and treated with dabsyl-chloride. The final HPLC separations are carried out on a reversed-phase column in a rapid isocratic run. The use of the cis isomer of hydroxyproline as an internal standard allows good reproducibility. As an application of the described method, the hydroxyproline content in samples containing collagen and an excess of bovine serum albumine (up to 20:1) is determined. PMID- 9358625 TI - Management of alloimmunized, refractory patients in need of platelet transfusions. PMID- 9358627 TI - The effect of different amines added to eluents as silanol masking agents on the chromatographic behavior of some diuretics in reversed-phase high-performance liquid chromatography using C18 packings. AB - A preliminary gradient separation in reversed-phase liquid chromatography of a mixture of 25 solutes (diuretics, probenecide, and atenolol) is carried out using several C18 columns and an aqueous phosphoric acid solution (pH 3.2)-acetonitrile mobile phase as a control. Using this separation, the chromatographic behavior of these solutes is studied using 11 water-soluble primary, secondary, and tertiary amine modifiers in the range of 0.7-7.5 mM and a Spherisorb C18 column. This study reveals the presence in the complex sample of two groups of solutes with positive (five typical solutes showed improvements in peak symmetry and retention) or negative responses using these amines as mobile phase modifiers. After experimentation in the presence of amines, these differences are related to solute structure. Hexylamine is found to be an effective masking agent of silanols because of its structure and small required concentration. On these bases, the silanophilic and hydrophobic character of typical solutes and several C18 packings are evaluated under isocratic elution and a relative effectiveness index for amines, and a method for their assessment is proposed. The role of the amine structure on solute retention and the importance of selecting amines of suitable hydrophobic character, molecular geometry, and concentration is discussed. A model of the formation and stabilization of the silanol-amine complex based on hydrophobic and ionic interactions is also proposed. PMID- 9358628 TI - Immunoaffinity chromatography in the detection of dexamethasone in equine urine. AB - Due to the widespread use of dexamethasone in racing horses, mostly in low doses by intra-articular administration for the treatment of inflammatory processes, a method is developed to detect this drug in horse urine samples using liquid liquid extraction followed by immunoaffinity chromatography. Liquid chromatography with diode-array detection is used for the identification of the drug. The use of immunoaffinity columns enhances the selectivity of the analysis, and the results show that dexamethasone can be detected up to 28 h after intra articular administration. PMID- 9358629 TI - Probing the substrate specificity of an enzyme catalyzing inactivation of streptogramin B antibiotics using LC-MS and LC-MS/MS. AB - LC-MS and LC-MS/MS analyses indicated that an enzyme responsible for inactivating the antibiotic etamycin is specific for streptogramins and acts on both type B-I and B-II streptogramin subgroups. No enzymatic activity was detected for other cyclodepsipeptides such as surfactins and viscosin. It was demonstrated using analogs of etamycin that the picolinyl moiety is essential to obtain enzyme generated ring-opened compounds. Because the picolinyl moiety is also essential for the biological activity of streptogramins, it is proposed that this residue is a distinctive topographic feature in the binding of this group of antibiotics to enzyme active sites. PMID- 9358631 TI - Delayed extraction time-of-flight MALDI mass spectrometry of proteins above 25,000 Da. AB - Matrix-assisted laser desorption/ionization (MALDI) with delayed extraction (DE) has been optimized for mass analysis of high-mass proteins and glycoproteins with masses above 25,000 Da. Under optimized experimental conditions, i.e. using a weak extraction field strength and a long delay time, a steep drop in mass resolution above 30,000 Da is no longer observed and an improvement of more than a factor of 10 is obtained compared with the non-DE case, at least up to 66 kDa in a 1.2 m time-of-flight mass analyzer. On this level of resolution the factors limiting further improvements become apparent, i.e. adduct ion formation between matrix and analyte, but also cationization and further non-matrix-related adducts, as well as prompt fragmentation. Moreover, heterogeneity of the sample is often the reason for the detection of broad signals for larger proteins. Within these limitations, DHBs (a 9:1 mixture of 2,5-dihydroxybenzoic acid and 2 hydroxy-5-methoxybenzoic acid) gave by far the best results. PMID- 9358630 TI - Determination of low 13C-glutamine enrichments using gas chromatography combustion-isotope ratio mass spectrometry. AB - Glutamine is an essential fuel for tissues with high rates of cell replication, such as enterocytes and lymphocytes. Infusion of 13C-labeled glutamine tracers allows for measurement of the rates of production, utilization and oxidation of glutamine's carbon skeleton in vivo. The use of this tracer, however, has been limited by its high cost and/or the difficulty is measuring low enrichments in biological fluids using conventional gas chromatography-mass spectrometry (GC/MS) techniques. We have developed a method using gas chromatography-combustion isotope ratio mass spectrometry (GC-C-IRMS) that allows for the determination of low 13C enrichments (down to 0.06 mol.% excess) with a precision of 2% or better, and a within-day and between-day variability better than 5%, in plasma free glutamine. The method was applied to measuring the incorporation of 13C in plasma glutamine over the course of infusion of 13C-labeled acetate in a human subject. PMID- 9358632 TI - The influence of strongly acidic groups on the protonation of peptides in electrospray MS. AB - In order to obtain experimental data on the question of compensation of positive charges by anionic groups in multiply charged ions of polyfunctional molecules in electrospray MS, several pairs of peptides with the same basic structure but differentiated by one or two strongly acidic groups (phosphate or sulfonate) were investigated. It was found that depending on the density of basic centers and the strength of acidic groups present, the observed changes upon introduction of acidic groups ranged from complete elimination of the most highly charged state to absence of any difference. PMID- 9358633 TI - Distinction between linear and angular 1,2-alkylenedioxy-benzofurazans and benzofuroxans by mass spectrometry. PMID- 9358634 TI - Analysis of Vero toxins 1 and 2 by high-performance liquid chromatography/electrospray ionization mass spectrometry. PMID- 9358635 TI - Isolation, structure elucidation, and biological activity of the steroid oligoglycosides and polyhydroxysteroids from the Antarctic starfish Acodontaster conspicuus. AB - A total of 19 steroids, of which 13 steroidal oligoglycosides (nine new and four known) and six polyhydroxylated steroids (four new and two known), has been isolated from the Antarctic starfish Acodontaster conspicuus. The mixture is dominated by glycosides composed of steroidal aglycons having the hydroxyl groups typically disposed on one side of the tetracyclic nucleus, i.e., 3 beta,4 beta,6 alpha,8,15 beta-, with some having a sulfate at C-6, and differing in the side chains and/or in the disaccharide moieties that are usually attached at C-26, with some at C-28 and C-29. Those compounds are accompanied by minute amounts of glycosides with a delta 8(14)-double bond in the steroid, which is a structural feature not previously found among polyhydroxysteroids derived from starfish. Small amounts of six related unglycosidated polyhydroxysteroids and three higher molecular-weight asterosaponins complete the composition of the mixture. The structures of the new compounds were determined by interpretation of their spectral data and by comparison with spectral data of known compounds. Eighteen of these compounds were evaluated for their ability to inhibit growth in Antarctic marine bacteria isolated from either the water column or the surfaces of benthic marine invertebrates. Of these compounds, 50% were active against at least one Antarctic marine bacterium. This suggests that these compounds may play an important role in deterring microbial fouling. PMID- 9358636 TI - Laurencia rigida: chemical investigations of its antifouling dichloromethane extract. AB - From the CH2Cl2 extract of the temperate marine red alga, Laurencia rigida, which has antifouling properties, eight sesquiterpenes (1-8) were isolated. Of these, four (3-acetoxy-E-gamma-bisabolene (1), (-)-10 alpha-bromo-9 beta-hydroxy-alpha chamigrene (2), rigidol (3), and (+)-(10S)-10-bromo-beta-chamigrene (4)), were shown to be new natural products. For the known compound deschloroelatol (5), reassignment of the 1H- and 13C-NMR data was found to be necessary on the basis of extensive NMR measurements. For elatol (6), complete 1H- and 13C-NMR data are also reported. The antimicrobial and antialgal activities of all isolates were assessed. PMID- 9358637 TI - Constituents of Dragon's Blood. 5. Dracoflavans B1, B2, C1, C2, D1, and D2, new A type deoxyproanthocyanidins. AB - From Dragon's Blood, a resin produced by plants of the genus Daemonorops (Palmae), six new A-type flavanoid deoxyproanthocyanidins have been isolated. Their structure and stereochemistry, established by chemical degradation and extensive NMR analysis, is consistent with a mechanism of formation common to other constituents of the resin, which involves oxidation of a 6-methylflavan to a quinonemethide, followed by coupling with another flavan moiety. PMID- 9358638 TI - New steroidal alkaloids from Buxus longifolia. AB - Four new steroidal alkaloids, (+)-cyclovirobuxeine F (1), N-benzoyl-O acetylbuxalongifoline (2), buxasamarine (3), and (+)-cyclobuxamidine (4), along with two known steroidal bases, 16 alpha-acetoxybuxabenzamidienine (5) and trans cyclosuffrobuxinine (6), were isolated from the leaves of Buxus longifolia. The alkaloids 1-4 showed significant antibacterial activity. PMID- 9358639 TI - Two new puriniums and three new pyrimidines from Heterostemma brownii. AB - Two new puriniums, heteromines D (4) and E (5), and three new pyrimidines, heteromines F (6), G (7), and H (8), were isolated from the aerial parts of Heterostemma brownii Hay. Their structures were determined as 7,9-dimethyl-2-(N,N dimethylamino)guaninium chloride, 7,9-dimethyl-2-(N-methylamino)guaninium chloride, 6-methoxy-4-(N-methylamino)-2-(N,N-dimethylamino)-5- (N methylformamido)pyrimidine, 6-methoxy-2,4-bis(N-methylamino)-5-(N methylformamido)pyrimidine, and 2-amino-6-methoxy-4-(N-methylamino)-5-(N methylformamido)-pyrimidi ne, respectively. PMID- 9358640 TI - Three new pseudodistomins, piperidine alkaloids from the ascidian Pseudodistoma megalarva. AB - Bioassay-guided fractionation of the MeOH-CH2Cl2 extract of the Micronesian ascidian Pseudodistoma megalarva yielded three new piperidine alkaloids, pseudodistomins D-F (3-5) and the previously reported pseudodistomins B and C (1 and 2). The structure and stereochemistry of these compounds were established by interpretation of spectral data. Pseudodistomins B-F were found to be active in a cell-based assay for DNA damage induction, but the activity was due to an alternative mechanism. PMID- 9358641 TI - Characterization of anticholinesterase-active 3-alkylpyridinium polymers from the marine sponge Reniera sarai in aqueous solutions. AB - From the marine sponge Reniera sarai 3-alkylpyridinium oligomers and polymers have been isolated. 3-Alkylpyridinium polymers are potent anticholinesterase agents; in addition, they show hemolytic and cytotoxic activities. Oligomers with a molecular weight lower than 3000 Da do not possess any significant activity. We report structural characterization of 3-alkylpyridinium polymers and their behavior in aqueous solutions. We found that biologically active polymers are composed of head-to-tail 3-alkylpyridinium units. According to MALDI-TOF spectrometry two species of polymers exist, the smaller with a molecular weight of 5520 Da and the larger with a molecular weight of 18,900 Da. Both polymers are soluble only in water, while low molecular oligomers are readily soluble in organic solvents. Polymers form large water-dissolved supramolecular structures with an average hydrodynamic radius of 23 +/- 2 nm and, therefore, cannot be separated with size-exclusion chromatography. PMID- 9358643 TI - Porrigenins A and B, novel cytotoxic and antiproliferative sapogenins isolated from Allium porrum. AB - Four new sapogenins, porrigenins A (2a) and B (3a), identified as (25R)-5 alpha spirostan-2 beta,3 beta,6 beta-triol and (25R)-2-oxo-5 alpha-spirostan-3 beta,6 beta-diol, respectively, and neoporrigenins A (2b) and B (3b) were also isolated from Allium porrum. In addition, the known agigenin (1a) and its 25S epimer, neoagigenin (1b), were also identified. Their structure elucidation was provided by comprehensive spectroscopic analyses. Compounds 1a, 2a, and 3a exhibited cytotoxicity and high antiproliferative activity on four different tumor cell lines in vitro. PMID- 9358642 TI - Antiandrogenic natural Diels--Alder-type adducts from Brosimum rubescens. AB - The isolation and structure elucidation of five novel natural Diels--Alder-type adducts, named palodesangrens A-E (1-5), from the Peruvian folk medicine known as "palo de sangre" (Brosimum rubescens) is described. The structures of the Diels- Alder adducts, consisting of chalcone derivatives and a prenylcoumarin, were elucidated by analysis of spectroscopic data including 2D NMR. Some of these compounds showed potent inhibitory activity towards 5 alpha-dihydrotestosterone (DHT) binding with an androgen receptor to form a DHT-receptor complex that causes androgen-dependent diseases. PMID- 9358644 TI - Bioactive constituents of Morus australis and Broussonetia papyrifera. AB - The biological activities of the active principles of two plants in the Moraceae have been investigated. A new prenylflavonoid, australone A (1), and a new triterpenoid, 3 beta-[(m-methoxybenzoyl)oxy]urs-12-en-28-ioc acid (2) were isolated from the root bark of Morus australis, and their structures determined by spectroscopic methods. Also isolated from this plant were seven known compounds, morusin (3), kuwanon C (4), betulinic acid, beta-amyrin, quercetin, ursolic acid, and compound A. Morusin (3) showed significant effects on arachidonic acid-, collagen-, and PAF-induced platelet aggregation, while kuwanon C (4) was active in the arachidonic acid- and PAF-induced platelet aggregation assays. In biological work on a second plant, Broussonetia papyrifera, broussoflavonols F (5) and G (6), broussoflavan A (7), and broussoaurone A (8) potently inhibited Fe(2+)-induced lipid oxidation in rat-brain homogenate. Compounds 5-7 also significantly inhibited the proliferation of rat vascular smooth muscle cells. PMID- 9358645 TI - Antiprotozoal compounds from Asparagus africanus. AB - Two antiprotozoal compounds have been isolated from the roots of Asparagus africanus Lam. (Liliaceae), a new sapogenin, 2 beta, 12 alpha-dihydroxy-(25R) spirosta-4,7-dien-3-one (1), which was named muzanzagenin, and the lignan (+) nyasol (2), (Z)-(+)-4,4'-(3-ethenyl-1-propene-1,3-diyl)-bisphenol. The structure of the sapogenin was elucidated by MS and by 1D and 2D NMR methods and established by a single crystal X-ray analysis. (+)-Nyasol potently inhibits the growth of Leishmania major promastigotes, the IC50 being 12 microM, and moderately inhibits Plasmodium falciparum schizonts with the IC50 49 microM. These concentrations only moderately affect the proliferation of human lymphocytes. Muzanzagenin showed a moderate in vitro activity in all three tests, the IC50 against leishmania promastigotes was 70 microM, and against four different malaria schizont strains the IC50 values were 16, 163, 23, and 16 microM, respectively. PMID- 9358646 TI - Biomimetic cyclization of cnicin to malacitanolide, a cytotoxic eudesmanolide from Centaurea malacitana. AB - Malacitanolide (2), a new eudesmanolide isolated from the aerial parts of Centaurea malacitana, was characterized spectroscopically. The synthesis of 2 from cnicin (1), via the epoxide 3, confirmed the structure and stereochemistry of malacitanolide, as well as its biogenetic relationship with 1. Cytotoxic activity values for 2 are significantly higher than for 1. PMID- 9358647 TI - Microbial transformation of sclareolide. AB - The microbial transformation of sclareolide (1) by Curvularia lunata afforded five oxidized metabolites identified as 3-ketosclareolide (2), 1 beta hydroxysclareolide (3), 3 beta-hydroxysclareolide (4), 1 alpha,3 beta dihydroxysclareolide (5), and 1 beta, 3 beta-dihydroxysclareolide (6). Fermentations of 1 with Aspergillus niger also produced metabolites 2-6. Metabolites 2-5 were obtained by fermention with Gibberella fujikuroii, while fermentation with Fusarium lini afforded metabolites 4 and 5. PMID- 9358648 TI - Plakinidine D, a new pyrroloacridine alkaloid from two ascidians of the genus Didemnum. AB - A previously undescribed red Didemnum sp. collected in Indonesia contained a novel pyrroloacridine, plakinidine D (4), along with the known compounds 3,5 diiodo-4-methoxyphenethylamine (5) and ascididemin (6), both of which had previously been isolated from ascidians of the genus Didemnum. Plakinidine D (4) and 3,5-diiodo-4-methoxyphenethylamine (5) were also isolated from Didemnum rubeum from the Republic of Palau. Interestingly, a collection of D. rubeum from Indonesia did not contain plakinidine D (4), but instead contained 3,5-diiodo-4 methoxyphenethylamine (5) and ascididemin (6). The structure of plakinidine D (4) was elucidated by analysis of its spectral data. Plakinidine D (4) is closely related to plakinidines A-C (1-3), previously isolated from the sponge Plakortis sp. PMID- 9358649 TI - Plakinidine D, a new pyrroloacridine alkaloid from the ascidian Didemnum rubeum. AB - Plakinidine D (4), a new pyrroloacridine metabolite, was isolated together with the known compound 3,5-diiodo-4-methoxyphenethylamine (5) from the ascidian Didemnum rubeum collected near the island of Rota, Northern Mariana Islands. Spectroscopic methods were used to determine the structure of plakinidine D, which was also confirmed through the formation and characterization of the derivatives N-acetylplakinidine D and 11-deoxyplakinidine D. Plakinidine D represents the first metabolite in the pyrroloacridine family of compounds to be isolated from an ascidian. PMID- 9358650 TI - A triterpenoid saponin from Patrinia scabiosaefolia. AB - A new triterpenoid saponin, patrinia saponin H3 (3), was isolated from the aerial parts of Patrinia scabiosaefolia Fisch. and determined to be 3-O-beta-D glucopyranosyl(1-->3)-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranosyl hederagenin 28-O-alpha-L-rhamnopyranosyl (1-->4)-beta-D-glucopyranosyl(1-->6) beta-D-glucopyranosyl ester on the basis of NMR and FAB MS experiments and acid hydrolysis. PMID- 9358652 TI - Study on the binding of cerium(III) ion and spermine to TRNA(Phe). AB - Fluorescence of Ce(III) aqua ion at pH 6.0 is found to be in good linear relationship with its concentration, and the intensity is strong enough to be employed to determine its concentration. TRNA(Phe) evidently quench this fluorescence. Fluorescence titration experiments were performed to Ce(III) tRNA(Phe) system, the binding number and association constant was estimated with a Scatchard plot. Two classes of binding sites with association constant of 5.2 x 10(7) and 4.4 x 10(6) M, respectively, were found. Addition of spermine slightly decrease binding number and association constant of Ce(III) ion. PMID- 9358651 TI - Red edge excitation shift emission spectroscopic investigation of serum albumins and serum albumin-bilirubin complexes. AB - The microenvironments around the intrinsic and extrinsic fluorophores in bovine and human serum albumins as well as their complexes with bilirubin have been visualized by red edge excitation shift (REES) emission spectroscopic investigation. The two albumins and their bilirubin complexes in aqueous buffered solutions (pH 7.5) do not exhibit any appreciable shift in their emission maxima, upon gradual change in excitation wavelength towards the red edge of their respective absorption band. The addition of Triton X-100 triggers REES emission in both the fluorophores. The observations suggest that the microenvironment around the flurophores are not so rigid, and even the extrinsic flurophore bilirubin having two carboxylic acid groups acts as a hydrophobic non-polar molecule when bound to albumins. The ligand binding domains (receptor sites) are large enough and incorporation of Triton X-100 makes the fluorophore environments rigid and subtle polarity may also be induced. Whereas small polar molecules like CHCl3, ANS and L-trp fail to induce REES emission in either of the fluorophores. PMID- 9358653 TI - Two-photon excitation of dioxane: time-resolved measurements of excited state complex formation with water. AB - We observed emission from the non-aromatic hydrocarbon 1,4-dioxane upon illumination with ps pulses at 380 nm. The emission intensity depended quadratically on incident power at 380 nm, indicating a two-photon process. In the absence of water the intensity decay was close to a single exponential, but displayed some evidence of an excited state process. In the presence of 1% water the emission spectra shifted dramatically to long wavelength. Water also resulted in wavelength-dependent intensity decays with negative pre-exponential factors on the long wavelength side of the emission, demonstrating the presence of an excited state reaction. At this water concentration the results are consistent with a two-state model due to emission from dioxane and a dioxane and a dioxane water complex. PMID- 9358654 TI - Cell activation influences cell staining kinetics. AB - Stimulation of cells has so far been observed, among other methods, by the decrease of the intracellular fluorescein fluorescence polarization (IFFP). It is shown that the rate constant of leakage of the fluorescent marker out of the cells increases with stimulation much more significantly than the polarization decreases; thus it might provide a more sensitive method to observe cells stimulation. It is also shown that due to negligible leakage of the marker out of the cells shortly after initiation of the staining of the cell suspension, the fluorescein fluorescence polarization (FFP) of the cell suspension, is very close to IFFP. PMID- 9358655 TI - Carboxyfluorescein as a fluorescent probe for cytoplasmic effects of lymphocyte stimulation. AB - The application of carboxyfluorescein (CF), as an impermeable fluorescent probe for lymphocyte stimulation with phytohaemagglutinin (PHA), is investigated by following a decrease in the degree of fluorescence polarization. Since CF does not enter the mitochondria, the present results indicate that the measured effect of stimulation occurs in the cytoplasm. The results also reveal that the fluorescence yield of intracellular CF is smaller than that of extracellular CF. Moreover, the degree of fluorescence polarization of intracellular CF is inversely related to its concentration. Following cell disruption, fluorescence intensity increases and polarization decreases. These effects might indicate a weak or reversible association of intracellular CF with cytoplasmic proteins. PMID- 9358656 TI - Conformational features of charged dibucaine in solution as investigated by 13C- and 1H-NMR. AB - Conformational features of charged dibucaine in [2H6]DMSO were elucidated by measuring 13C and 1H spin-lattice relaxation rates and 1H-(1H) and 13C-(1H) nuclear Overhauser effects. The reorientational correlation time of the aromatic moiety was evaluated at 0.16 ns at room temperature and side chains were observed to display segmental motion. Relevant distances were calculated by isolating dipolar interaction terms of 1H-1H or 13C-1H pairs. The 'preferred' conformation in solution was shown to present several analogies, but also some differences, with the structures obtained by solid state experiments, energy calculations and LIS data. PMID- 9358657 TI - Expression and characterization of the recombinant gene encoding chitinase from Aeromonas caviae. AB - The gene encoding a chitinase from Aeromonas caviae was cloned by PCR techniques. Its recombinant gene expression was performed using pET20b(+) in Escherichia coli BL21 (DE3). The recombinant chitinase with the extra 33 and 13 amino acids in its N- and C-termini, respectively, was purified to near homogeneity using His-Tag affinity chromatography. The recombinant chitinase was found to be present in both the culture medium and the cytoplasm. A single protein band on the native polyacrylamide gel was confirmed by both the activity staining and protein staining. The optimum pH and temperature of the recombinant chitinase were determined to be 6.25-6.5 and 42.5 degrees C, respectively. It was stable within the pH range of 5-7. Significant activity stimulation by Cu2+ and inhibition by Fe3+ and Hg2+ were observed. Detergents such as SDS and Triton X-100 strongly inhibited the enzyme activity. Substrates such as 4-methylumbelliferyl-N,N' diacetylchitobioside and 4-methylumbelliferyl-N,N',N"-triacetylchitotriose were hydrolyzed by the recombinant chitinase; however, 4-methylumbelliferyl-N acetylglucosaminide was not cleaved during the activity assay periods. When chitin power was suspended in buffer with the chitinase (pH 6.5 and 42.5 degrees C), N-acetylchitooligosaccharides [(GlcNAc)n, n = 1-4] were detected at 24 h. PMID- 9358658 TI - The WHO/WHL patient education project "Teaching the Teacher". PMID- 9358659 TI - Therapeutic issues in prosthetic heart valve replacement. PMID- 9358661 TI - Is haemorrhagic myocardial infarction more common with streptokinase? AB - Autopsy reports and clinical data of 226 consecutive myocardial infarction deaths in whom postmortem studies could be carried out during the period 1980 to 1996 were analyzed retrospectively for the presence of haemorrhagic myocardial infarction (HMI). Of 53 autopsies done from 1980 to 1986 [prior to use of streptokinase (SK) therapy in our institution] none of the specimens showed haemorrhagic infarction. Of 173 autopsies done from 1987 to 1996 (intravenous SK therapy was utilised in this period), 20 specimens showed haemorrhagic infarctions. Sixteen of these 20 patients had received SK, while 66 of the remaining 153 non-haemorrhagic myocardial infarction patients received SK (statistically significant association of SK with HMI, p < 0.005). Acute mechanical complications [ventricular septal rupture (n = 10), papillary muscle rupture (n = 2), cardiac free wall rupture (n = 7)] were seen in 19 cases. Of these, 16 were HMIs and 14 of these patients had received streptokinase. These observations suggest a strong association of HMI with SK therapy and with acute mechanical complications. PMID- 9358660 TI - Thrombolytic therapy for prosthetic valve thrombosis in Third World countries. AB - In order to clarify the role of thrombolytic therapy for treatment of prosthetic valve thrombosis, all cases admitted in the intensive care unit (ICU), between March 1987 and March 1997 with the diagnosis of prosthetic valve thrombosis and treated with streptokinase, were analysed. In total, 42 patients with clinical and echocardiographic evidence of left side tilting disc prosthetic valve thrombosis were treated. All the patients had only mitral valve prosthesis involvement. Streptokinase was administered as a bolus of 2.5 lac units over 30 minutes followed by 1 lac units/hour for 48-72 hours. Thirty-seven (88%) patients had successful thrombolysis. Overall mortality occurred in 9.5 percent patients due to systemic embolism and bleeding complications. Serial clinical, radiological and echocardiographic studies showed successful thrombolysis in 88 percent patients. This study demonstrates that streptokinase therapy is safe and effective first line treatment for left-sided prosthetic valve thrombosis and surgery should be reserved for those patients who fail to respond to thrombolytic therapy. PMID- 9358662 TI - Immediate and follow-up clinical outcome after multivessel coronary stenting. AB - Seventy-two out of 656 patients treated by coronary stenting between January 1995 to May 1997 underwent elective multivessel stenting as a strategy for nonsurgical revascularization in patients with two-vessel (n = 37) and three-vessel (n = 35) disease. Their age ranged from 35 to 77 years (mean: 53.6 +/- 9.2) and the majority (77.8%) were males. The patients were included if the target vessel was more than 2.7 mm in diameter and subserved a moderate to large area of viable myocardium, provided the target lesion was considered approachable by stent. In all, 160 stents were deployed in 146 vessels with a mean of 2.2 stents per patient. The procedure was performed on all the target lesions in one stage in 51(70.8%) and two stages in 21(29.2%) patients. Two-vessel stenting was done in all except 2 patients who received stents in all the three major arteries. Successful deployment of the stent was achieved at the target site in all patients without any major in-hospital complications including subacute stent thrombosis, myocardial infarction (MI), emergency bypass graft surgery (CABG) or death. Clinical follow-up was available in 66(91.6%) patients at a mean of 7.8 +/ 5.5 months. The actuarial survival rates were 98.6, 96.7 and 94.6 percent, respectively at one, 3 and 6 to 12 months after the procedure with an event-free survival (absence of death, MI, recurrence of angina or any revascularization) of 98.5 percent at one, 93 percent at 3, 83.2 percent at 6 and 68.4 percent at 12 months. Only 15(22.7%) patients developed any event and target lesion revascularization was required in 8(12%) patients. In conclusion, multivessel stenting in patients with two- and three-vessel coronary disease is feasible, safe and effective in preventing major in-hospital complications as well as reducing the recurrence of clinical events and need for revascularization on follow-up. PMID- 9358663 TI - Analysis of local electrograms at successful and unsuccessful sites for slow pathway ablation of atrioventricular nodal reentrant tachycardia. AB - Anatomic and electrogram approaches have been described for ablation of slow pathway in patients with atrioventricular nodal tachycardia. The purpose of this study was to identify parameters to predict successful slow pathway ablation using the anatomic approach. Local electrograms at successful and unsuccessful sites were compared in 36 patients undergoing slow pathway ablation using anatomic approach. A total of 208 local electrograms were studied. Fragmented atrial electrogram was seen in 24/36 (67%) of successful and in 46/172 (26%) of unsuccessful sites (p < 0.001). The sensitivity, specificity and positive and negative predictive values of fragmented atrial electrogram were 67, 73, 34 and 91 percent respectively. A slow pathway potential was noted in three of successful sites. There was no difference in the atrial to ventricular amplitude ratio in these sites. In conclusion, fragmentation of atrial electrogram and presence of possible slow pathway potential are seen more often at successful than at unsuccessful sites. In our opinion, while using an anatomic approach for slow pathway ablation, an analysis of local electrogram may help in identifying the proper site and avoiding unnecessary radiofrequency energy delivery. PMID- 9358664 TI - Inappropriate shocks in patients receiving internal cardioverter-defibrillators for malignant ventricular arrhythmias. AB - Implantation of internal cardioverter defibrillators (ICDs) for treatment of malignant ventricular arrhythmias is complicated by failure of therapy or inappropriate shocks. We studied 81 patients (age range 16-72 years; mean 48 +/- 13 years) who underwent ICD implantation for device therapy. The underlying aetiology was ischaemic heart disease (39%), cardiomyopathies (32%) and others (28%). Information regarding shocks was collected using Holter monitoring, telemetry or device memory (stored electrograms) and lastly by clinical follow up. Fifty-eight patients completed 36 months of follow-up. Thirty-five patients experienced 337 spontaneous shocks, appropriate in 21, inappropriate in 12, and both in two patients. Of the 74 episodes of inappropriate discharges for rhythms other that ventricular tachycardia (VT) or ventricular fibrillation(VF), 55 percent were due to supraventricular arrhythmias (atrial flutter or fibrillation). Lead malfunction occurred in four and the device was replaced in two. Additional drugs controlled AF in one. There was no mortality in any of the 81 patients. The frequency of shocks was highest in the first six months after implantation and atrial fibrillation remains the main cause. In conclusion, inappropriate shocks are frequent in patients undergoing ICD implantation. PMID- 9358665 TI - Evaluation of left ventricular functions in chronic renal failure before and after acute hemodialysis. AB - Left ventricular functions were evaluated in 25 adult patients of chronic renal failure by 2-D echocardiography before and after four hours of standard hemodialysis session. Eighteen patients showed clinical evidence of fluid overload. Predialysis left ventricular end-diastolic diameter, left ventricular end-diastolic volume, left ventricular end-systolic diameter and left ventricular end-systolic volume were comparable in patients with or without fluid overload. Similarly, predialysis stroke volume and left ventricular ejection fraction were not significantly different in the two subsets. However, following hemodialysis there was a significant decrease in the left ventricular systolic and diastolic volumes and diameters in patients with fluid overload. The improvement in the left ventricular ejection fraction was of the same magnitude in the two subsets. The significant improvement in the left ventricular functions both in patients with and without fluid overload indicates that fluid overload may not be the only determinant of left ventricular functions in patients of chronic renal failure, but other factors, such as various uraemia toxins and metabolic changes might also be inhibiting the myocardial functions. PMID- 9358666 TI - Antitubercular treatment does not prevent constriction in chronic pericardial effusion of undetermined etiology: a randomized trial. AB - Patients of chronic exudative pericardial effusion are frequently treated with antitubercular treatment on presumptive grounds in developing countries, in a hope to prevent constrictive pericarditis. To assess the impact of antitubercular treatment on development of constrictive pericarditis in chronic large exudative pericarditis effusion of undetermined etiology, 25 patients above 12 years of age, with large pericarditis effusion beyond 12 weeks duration, were randomized in a prospective 2:1 fashion, to receive either 3-drug antitubercular treatment (group A) or placebo (group B) for six months. End points studied were, development of pericardial thickness as diagnosed by CT scan and constrictive pericarditis as diagnosed by cardiac catheterization. Twenty-one patients (14 in group A and 7 in group B) completed the study protocol. In all, five (23.8%) patients developed constrictive pericarditis/pericardial thickening. Histopathological examination of pericardiectomy specimens in over five patients were negative for tubercular pathology. Pericardial effusion resolved completely in another 10 (47.8%) patients. There was no significant difference in both the groups in development of constrictive pericarditis/pericardial thickening (group A: n = 3, 21.4% and group B: n = 2, 29.6%, p = NS). On multivariate analysis, development of constrictive pericarditis/pericardial thickening was associated with recurrent tamponade (p = 0.01), presence of tamponade at admission (p = 0.07) and haemorrhagic pericardial effusion (p = 0.08). Thus, antitubercular treatment does not prevent the development of constrictive pericarditis in patients of large chronic pericardial effusion of undetermined etiology. PMID- 9358667 TI - Familial Lutembacher's syndrome in mother and daughter. PMID- 9358668 TI - Double outlet left atrium. PMID- 9358669 TI - Complete common AV canal with long survival. PMID- 9358670 TI - Left atrial abscess with mitral stenosis and peripartum cardiomyopathy. PMID- 9358671 TI - Combined percutaneous coronary and carotid artery stenting. PMID- 9358672 TI - Percutaneous balloon dilatation of IVC obstruction using Inoue catheter. PMID- 9358673 TI - Balloon angioplasty followed by balloon mitral valvotomy and renal transplantation in a patient of end stage renal disease. PMID- 9358674 TI - Pregnancy with type II aortoarteritis and descending aortic occlusion. PMID- 9358676 TI - The tyranny of prevailing opinions. PMID- 9358675 TI - Antiarrhythmic drug trials: what lessons have we learned? PMID- 9358677 TI - Ethical concerns in modern medical practice. PMID- 9358678 TI - Meta-analysis of prevalence of hypertension in India. PMID- 9358679 TI - Haemorrhage resulting in isolated myocardial infarction without coronary atherosclerosis in a young lady. PMID- 9358680 TI - The prevalence and biometric structure of pathological dissociation in the general population: taxometric and behavior genetic findings. AB - Taxometric and biometric analyses were conducted on 2 North American samples to investigate the prevalence and biometric structure of pathological dissociation. Results indicated that approximately 3.3% of the general population belongs to a pathological dissociative taxon. A brief 8-item self-report scale called the DES T can be used to calculate taxon membership probabilities in clinical and nonclinical samples of adults (a SAS scoring program is provided for this purpose). The genetic and environmental architecture of pathological dissociative symptoms was explored by conducting a biometric analysis on DES-T ratings from 280 identical and 148 fraternal twins. The findings suggest that approximately 45% of the observed variance on the DES-T can be attributed to shared environmental influences. The remaining variance is due to nonshared environmental influences. PMID- 9358681 TI - Thoughts of agoraphobic people during scary tasks. AB - The authors examined the occurrence of theoretically derived patterns of thinking in 74 agoraphobic participants as they drove alone or tolerated an enclosed place. During the increasingly scary tasks in a behavioral test hierarchy, participants responded to a periodic beep by stating aloud what they were thinking at that moment, yielding more than 1,800 tape-recorded statements. Content analyses revealed that participants were mainly preoccupied with their current anxiety (expressed in 29% of the statements) and with their self-efficacy (15%). Despite participants' mounting feelings of anxiety, fewer than 1% of their statements expressed a thought of danger or an anticipation of future anxiety or panic. The rarity of danger thoughts poses an explanatory challenge for all cognitive theories of phobia and especially for the perceived danger theory of A. T. Beck (1976) and A. T. Beck, G. Emery, and R. L. Greenberg (1985). PMID- 9358682 TI - Temporal variability in global self-esteem and specific self-evaluation as prospective predictors of emotional distress: specificity in predictors and outcome. AB - Recent studies have found that temporal variability and reactivity in self-esteem (SE) are associated with risk for depressive symptoms subsequent to life stress. It is unclear, however, whether it is variability uniquely in SE that is critical, or whether variability in other domains, such as specific self evaluation (SSE) and affect, would show similar effects. Further, the specificity of these effects to depression is unknown. In the present study, initially nondepressed women completed 7 daily ratings of SE, SSE, and affect. Over a 6 week prospective interval, the interactions of stressful life events and variability in both SE and SSE predicted changes in depression, particularly in individuals with more severe worst lifetime episodes of depressive symptoms. These effects were independent of average level of SE and SSE, as well as neuroticism and self-concept uncertainty. In contrast, variability in affect failed to predict changes in depression in interaction with life stress. Finally, none of the predictor variables interacted with stressful life events in predicting changes in anxiety. PMID- 9358683 TI - Perceptual organization in schizophrenia: utilization of the Gestalt principles. AB - In 2 experiments, the authors investigated perceptual organization in schizophrenia to determine whether patients with schizophrenia used the Gestalt principles. In the visual embedded figures task (Experiment 1), the authors examined the principles of proximity and collinearity, whereas in the similarity task (Experiment 2), the authors examined similarity. Forty-three people participated in the study: patients diagnosed with schizophrenia (14) or with bipolar disorder (15), and "normal" participants (14). All 3 groups of participants showed performance superiority in the condition that was facilitated by the use of the Gestalt principles in both tasks. This study supports the hypothesis that visual perceptual organization, in terms of utilizing the Gestalt principles, is relatively intact in people with schizophrenia. PMID- 9358684 TI - The relation of anxiety sensitivity to higher and lower order personality dimensions: implications for the etiology of panic attacks. AB - The relation of anxiety sensitivity (AS) to personality dimensions has received little attention. In this study, 4 AS indexes were administered along with measures of personality, fears, and panic attacks to 220 undergraduates. At the higher order level, AS was positively correlated with negative emotionality (NE) but was largely unrelated to either positive emotionality or constraint. At the lower order level, AS was positively correlated with absorption and NE indexes. Most of these correlations were significant even among participants with no panic attack history. AS exhibited incremental validity above and beyond a number of personality variables, including absorption and trait anxiety, in the prediction of fears and panic attack history. These findings are consistent with the hypothesis that a propensity toward immersion in sensory experiences is a diathesis for panic attacks. PMID- 9358685 TI - Personality, temperament, and character dimensions and the DSM-IV personality disorders in substance abusers. AB - The authors evaluated the relationship between P. T. Costa and R. R. McCrae's (1992) NEO 5-factor model, C. R. Cloninger's (1993) 7-factor Temperament and Character Inventory (TCI), and the American Psychiatric Association's (1994) Diagnostic and Statistical Manual of Mental Disorders, 4th ed., personality disorders in 370 inpatient and outpatient alcohol, cocaine, and opiate abusers. NEO Neuroticism was associated with many disorders, and different patterns for Agreeableness, Conscientiousness, and Extraversion emerged for the different disorders. Several TCI scales were associated with different personality disorders, although not as strongly as the NEO dimensions. Results did not support most predictions made for the TCI. Normal personality dimensions contributed significantly to the prediction of personality disorder severity above and beyond substance abuse and depression symptoms. PMID- 9358687 TI - The impact of motivationally neutral cues on psychopathic individuals: assessing the generality of the response modulation hypothesis. AB - Psychopathic individuals' lack of responsiveness to punishment cues and poor self regulation have been attributed to fearlessness (D. T. Lykken, 1957, 1982, 1995). Alternatively, deficient response modulation (RM) may hinder the psychopathic individual's processing of peripheral information and self-regulation when they are engaged in goal-directed behavior (C. M. Patterson & J. P. Newman, 1993). Although more specific than the fearlessness hypothesis in some respects, the RM hypothesis makes the more general prediction that psychopathic individuals will have difficulty processing motivationally neutral as well as fear-related stimuli. The authors assessed this prediction by using psychopathic and nonpsychopathic male inmates subdivided by level of anxiety/negative affectivity (NA). As predicted by the RM hypothesis, peripheral presentation of motivationally neutral cues produced significantly less interference in low-NA psychopathic individuals than in low-NA controls. PMID- 9358686 TI - Which environmental variables are related to the onset of seasonal affective disorder? AB - The regular fall-winter onset of seasonal affective disorder is believed to be related to seasonal changes in the environment. However, the high correlation among various environmental variables has made it difficult to distinguish which ones may play a causal role. Photoperiod should explain variations in onset risk across both latitude and day of the year because it varies as a function of only these 2 factors. In Study 1, the authors found this to be the case using data from 5 locations. Environmental factors that vary from year to year should explain variations in onset risk across both time of year and actual year. In Study 2, the authors examined data from 7 years at 1 location and failed to find evidence of this effect for daily hours of sunshine, mean daily temperature, and total daily radiation. Findings support photoperiod as being related to the onset of seasonal affective disorder. PMID- 9358688 TI - Construct validity of psychopathy in a female offender sample: a multitrait multimethod evaluation. AB - The authors examined the construct of psychopathy as applied to 103 female offenders, using the multitrait-multimethod matrix proposed by D. T. Campbell and D. W. Fiske (1959). Instruments used in the study included the following: (a) Antisocial Scale of the Personality Assessment Inventory (L. C. Morey, 1991); (b) Psychopathy Checklist--Revised (R. D. Hare, 1990); and (c) Antisocial scale of the Personality Disorder Examination (A. W. Loranger, 1988). Criterion-related validity was also evaluated to determine the relationship between psychopathy and staff ratings of aggressive and disruptive behavior within the institution. Results revealed significant convergence and divergence across the instruments supporting the construct of psychopathy in a female offender sample. The measures of psychopathy demonstrated moderate convergence with staff ratings of violence, verbal aggression, manipulativeness, lack of remorse, and noncompliance. It is interesting to note that an exploratory factor analysis of the PCL-R identified a substantially different factor structure for women than has been previously found for male psychopathy. PMID- 9358689 TI - Cumulative and compensatory effects of competence and incompetence on depressive symptoms in children. AB - The authors tested 5 hypotheses from a competency-based model of child depression using classification and regression tree analysis. The authors obtained measures of 5 domains of competency (i.e., academic competence, social acceptance, sports competence, physical attractiveness, and behavioral conduct) and depressive symptoms that were derived from parent, teacher, peer, and self-reports on 1,063 3rd- and 6th-grade children. Results suggested that (a) multiple domains of competence related to depressive symptoms, (b) significant others' positive evaluations in multiple domains have a cumulative inverse relation to depressive symptoms, (c) negative evaluations in multiple domains have a cumulative but positive relation to depressive symptoms, (d) positive evaluations in one domain somewhat compensate for negative evaluations in another domain, and (e) negative evaluations in one domain offset positive evaluations in another domain. PMID- 9358691 TI - Self-verification and depression among youth psychiatric inpatients. AB - According to self-verification theory (e.g., W.B. Swann, 1983), people are motivated to preserve stable self-concepts by seeking self-confirming interpersonal responses, even if the responses are negative. In the current study of 72 youth psychiatric inpatients (36 boys; 36 girls; ages 7-17, M = 13.18; SD = 2.59), the authors provide the 1st test of self-verification theory among a youth sample. Participants completed self-report questionnaires on depression, self esteem, anxiety, negative and positive affect, and interest in negative feedback from others. The authors made chart diagnoses available, and they collected peer rejection ratings. Consistent with hypotheses, the authors found that interest in negative feedback was associated with depression, was predictive of peer rejection (but only within relatively longer peer relationships), was more highly related to cognitive than emotional aspects of depression, and was specifically associated with depression, rather than being generally associated with emotional distress. The authors discuss implications for self-verification theory and for the phenomenology of youth depression. PMID- 9358690 TI - The biphasic effects of alcohol on human physical aggression. AB - The authors assessed the biphasic effects of alcohol on human physical aggression. Sixty male social drinkers were assigned to 1 of 4 groups: alcohol ascending limb (AAL), alcohol descending limb (ADL), or 1 of 2 sober control groups. Aggression was assessed in the AAL and ADL groups at respective ascending or descending blood alcohol concentrations (BAC) of 0.08%. Each participant in the control groups was respectively yoked with a participant in either the AAL or the ADL group to control for the longer period of time needed to reach a BAC of 0.08% on the descending limb compared with the ascending limb (i.e., passage of time effect). The authors measured aggression using a modified version of the Taylor aggression paradigm (S. Taylor, 1967), in which electric shocks are received from and administered to a fictitious opponent during a competitive task. The AAL group was more aggressive than the ADL groups. There were no differences between the ADL group and the control groups, which suggests that alcohol does not appear to increase aggression on the descending limb. The control groups did not differ in aggression, thus ruling out a passage of time effect. PMID- 9358692 TI - Predicting stability and change in frequency of intoxication from the college years to beyond: individual-difference and role transition variables. AB - The authors examined whether individual-difference variables (e.g., family history of alcoholism, sex, personality traits, positive alcohol expectancies) and role transition-related variables (full-time work status, marital status, parenthood) moderate the "maturing-out" process whereby young adults who drink heavily during college decrease their drinking in the following years. Analyses were based on 288 young adults, assessed as full-time students (mostly college seniors, Year 4 of a larger study) and 3 years later (Year 7) when all had earned bachelor's degrees, and the analyses showed that frequency of intoxication (per week) decreased significantly (p < .0001). Entering the workforce full time, being male, and being less open to experience were associated with decreased postcollege drinking. Furthermore, relatively extraverted individuals were more likely to continue a pattern of frequent intoxication from Year 4 to year 7. The findings stress the importance of studying how individual-difference variables predict behavior across role transitions. PMID- 9358693 TI - Extinction of panicogenic effects of a 35% CO2 challenge in patients with panic disorder. AB - Inhalations of high concentrations of carbon dioxide (CO2) reliably produce panic attacks in patients with panic disorder. The present study evaluated whether cognitive-behavioral treatment (CBT) for panic disorder would extinguish CO2 induced panic and whether changes in panic and arousal-related cognitions were associated with the induction of panic. Patients with panic disorder (N = 54) were assigned to 1 of 3 experimental conditions: CBT with respiratory training (CBT-R), CBT without respiratory training (CBT), or delayed treatment. Participants received 5 repeated vital-capacity inhalations of 35% CO2/65% O2 prior to and following either 12 treatment sessions or a 12-week waiting period. During pretreatment assessments, 74% of patients experienced a panic attack during at least 1 inhalation. At posttreatment, only 20% of treated participants (CBT-R = 19%, CBT = 22%), compared with 64% of untreated participants, panicked. Forty-four percent of treated participants, compared with 0% of untreated participants, reported no anxiety during all posttreatment inhalations. Anxiety sensitivity as well as panic appraisals regarding the likelihood of panic and self-efficacy with coping with panic were significantly related to fearful responding to the CO2 challenge. PMID- 9358694 TI - The effects of antipsychotic medication on sequential inhibitory processes. AB - The authors used a Stroop negative priming paradigm to examine the effects of antipsychotic medication on selective attentional processes. The performance of 14 patients with schizophrenia who were withdrawn from neuroleptic medication was compared with that of 10 medicated patients and 16 matched controls. Results demonstrated an increase in negative priming to normal levels with neuroleptic therapy. In contrast, within-trial interference and facilitation effects appeared to be less sensitive to medication therapy. The sustainment of inhibitory processes over time may differentiate the inhibitory mechanisms of the medication withdrawn patients from both the medicated patients and the matched controls. The study of sequential inhibitory processes and their response to neuroleptic treatment could be important methods for understanding the temporal parameters associated with inhibition in schizophrenia. PMID- 9358695 TI - Phobia-related cognitive bias for pictorial and linguistic stimuli. AB - The purpose of this study was to examine whether anxiety-related cognitive bias for threat is stronger for threatening pictures than for threatening words. Spider-phobic participants (n = 31) and control participants (n = 33) performed a pictorial and linguistic spider Stroop task. Spider-phobic participants showed a marked bias for threat. However, this bias was similar for pictures and for words, although the spider-phobic group evaluated the pictures as being more aversive. The results suggest that automatic processing of threatening information in people with phobias is triggered in an on-off fashion, independent of subjective threat of the stimuli. This lack of distinction in automatic processing of weak and strong predictors of danger may be fundamental to the irrational nature of anxiety disorders. PMID- 9358696 TI - Schizotypy in community samples: the three-factor structure and correlation with sustained attention. AB - The authors examined cross-cultural applicability of the Schizotypal Personality Questionnaire (SPQ), the Perceptual Aberration Scale (PAS), and the Continuous Performance Test in community samples of Taiwanese adults and adolescents. The authors tested hypotheses that a 3-factor structure of the SPQ (Cognitive Perceptual, Interpersonal, and Disorganization: A. Raine et al., 1994) exists for both samples and that the Interpersonal factor is associated with poorer attention. The authors replicated the 3-factor model for both samples, and they externally validated the model in adults: The Interpersonal factor and possibly the Disorganization factor were associated with poorer attention, whereas the Cognitive-Perceptual factor was not. The PAS differed from the Cognitive Perceptual factor in its consistent association with poorer attention in both samples. These have important implications for the scales in the early detection of the schizotypy in various cultures. PMID- 9358697 TI - The Core Competencies for Basic Midwifery Practice. Critical ACNM document revised. American College of Nurse-Midwives. PMID- 9358698 TI - The Core Competencies for Basic Midwifery Practice. Adopted by the American College of Nurse-Midwives. May 1997. PMID- 9358699 TI - Epidural anesthesia in labor. Benefits versus risks. AB - Epidural anesthesia is used for relief of labor pain by 29% of women having hospital deliveries in the United States, a number that has doubled within the past 10 years. Although epidurals provide objective pain relief that is exponentially better than the other pain relief methods, there are many purported complications and side effects. This article reviews how epidurals work, summarizes the literature regarding complications, and presents some of the ethical dilemmas inherent in the use of this technology for labor. PMID- 9358700 TI - Informed consent for epidural analgesia in labor. AB - Epidural analgesia is widely available and increasingly popular in the United States for pain relief in childbirth. Although it provides superior pain relief for most women, it is not without significant short- and long-term side effects. It is costly and requires the use of numerous other technologic interventions. Because women have information needs surrounding childbirth and value self determination, full informed consent regarding epidural analgesia should be a priority in patient care. The best time and place in which to accomplish this is during the prenatal period. PMID- 9358701 TI - Macroscopic examination of the placenta immediately following birth. AB - Macroscopic examination of the placenta can be performed readily by the healthcare providers present at the time of birth. It can be accomplished with a basic knowledge of placental pathology and a general understanding of the abnormalities and variations that affect the placenta. These observations can easily be entered into the medical record and may often provide information about intrauterine life that will help the future care of both mother and infant. PMID- 9358702 TI - Clinical bulletin No. 2-January 1997. Early-onset group B strep infection in newborns prevention and prophylaxis. AB - A systematically applied strategy for selecting candidates for intrapartum chemoprophylaxis to decrease the incidence of early-onset GBS disease is an important component of midwifery care. The strategy that an individual practice chooses to follow will depend upon multiple factors including but not limited to: the prevalence of GBS colonization in the prenatal population, ability of clients to return for regular prenatal visits, availability of laboratory services, choices made for screening and treatment by collaborating obstetric and pediatric colleagues, place of birth and logistics required for protocol compliance. PMID- 9358703 TI - Risks of devices for direct detection of group B streptococcal antigen. PMID- 9358704 TI - Patient preference for self-collected cultures for group B streptococcus in pregnancy. AB - To determine pregnant women's preference for self-culture technique, 251 women between 24 and 42 weeks' gestation were interviewed after performing self collected cultures (vaginal and rectal) for group B streptococcus. Patient receptiveness to self-culture, the ability to perform self-culture, and the desire for choice in the future were derived using the Patient Preference Tool. The majority of women (77%, n = 194) gave positive descriptions of self-culture technique, and the majority of women preferred self-culture technique over nurse collected sampling (57%, n = 142). Seventy-nine percent (n = 197) stated their desire to have a choice about self-culture in the future when similar testing was needed, and 89% (n = 224) believed that other women would also like this choice. Additionally, patient samples were highly correlated with nurse-collected samples for accuracy of culture results. This study provides data supporting that women desire active participation in their care. PMID- 9358705 TI - Can water immersion stop labor? AB - The basic response to any sort of immersion is a redistribution of blood volume. The intrathoracic blood volume expansion is associated with an increased release of the Atrial Natriuretic Peptide (ANP) and an alteration of the activity of the posterior pituitary gland. Whereas the effects of ANP on vasopressin secretion have been studied extensively, there is little study of its effects on oxytocic release. This is one reason why a dialogue is needed between midwives and specialized physiologists regarding the relationship between water immersion and the physiology of labor. PMID- 9358706 TI - Midwifery as a discipline. AB - Recent developments within midwifery necessitate an examination of whether midwifery qualifies as a discipline as well as a profession. A discipline is a formalized branch of knowledge and is different from, although related to, a profession of the same name. The author contends that the discipline of midwifery is undeveloped at present but that the time is right for explicit discussion about what defines the discipline of midwifery. PMID- 9358707 TI - Trends in state laws and regulations affecting nurse-midwives: 1995-1997. AB - This article provides an overview of trends and changes to state laws and regulations governing the practice of nurse-midwifery in the United States from 1995 to 1997. It includes tables documenting state requirements related to education, certification, Medicaid and third-party reimbursement, prescriptive, authority, graduate practice, birth centers, regulatory oversight and postpartum discharge. PMID- 9358708 TI - The University of California, San Diego Nurse-Midwifery Bridge Program. An opportunity to learn intrapartum nursing skills. AB - This article describes an innovative program developed by the University of California, San Diego Nurse-Midwifery Education Program to supplement the experiential background of prospective nurse-midwifery students. This preprogram enables RNs without previous labor and delivery (maternal/newborn) nursing experience to complete the bridge program and enter the UCSD student nurse midwifery program on the same clinical level as other students and complete it within the standard time frame. The rationale for the program, its structural framework, and evaluation tools are presented. The authors hope that a description of the program will allow other institutions to replicate UCSD's success. PMID- 9358709 TI - Direct-entry midwifery education: history in the making. PMID- 9358710 TI - Grappling with the dilemma of whether to support the education of non-nurse midwives. PMID- 9358711 TI - What's in a name: defining the profession. PMID- 9358713 TI - Therapeutic intervention on four paws. PMID- 9358712 TI - It is likely that CNM's prescriptive privileges will, in the near future, cover mifepristone and misoprostol. PMID- 9358714 TI - A four-year review of patients with hepatitis C antibody in Department of Veterans Affairs facilities. AB - Hepatitis C virus [HCV] has recently been recognized as an emerging pathogen of surprising proportions. The clinical liver illness associated with HCV infection can be minimal or none, but in a notable number of persons it can ultimately cause debilitating chronic liver disease, fibrosis, cirrhosis, and hepatic failure, and it is likely related to an increased incidence of hepatocellular carcinoma. From 1991 to 1994, an annual electronic census was sent to 168 Veterans Health Administration facilities requesting serologic data on HCV in patients seen in Department of Veterans Affairs facilities. Response rate to the mandatory survey was 100%. In 1991, 6,612 individual patients were reported as positive for HCV antibody in the Department of Veterans Affairs system. This increased yearly from 1992 to 1994 with 8,365, 14,097, and finally 18,854 persons, respectively. This represents an increase of more than 285% during the 4 year period. Increases in HCV antibody for the same period were seen in all major regions of the United States and in the specified large metropolitan areas. In the New York area and in coastal California, this trend of new case identification may have plateaued during 1993 and 1994. Overall, total patients seen nationally in the Veterans Health Administration increased by only 4.87% during the same period, 1991 to 1994. Thus, the increase in persons positive for HCV antibody is not based on work load alone. The impact of HCV disease on patient well-being and health care costs cannot be overestimated. PMID- 9358715 TI - Smoking cessation in military personnel. AB - Tobacco use is the single most important preventable cause of death in military personnel. The purpose of this randomized clinical trial was to evaluate the effectiveness of two behavioral interventions when added to nicotine-replacement therapy on smoking cessation. The sample of 512 included 52% active duty military, 29% family, 11% retirees, and 8% Department of Defense civilians. There was a main effect of compliance at the end of the program (EOP); 69% of those who attended 75% of the classes were abstinent from tobacco; regression analysis found the more intensive program to be twice as effective at EOP and at 3 months, an outcome not continued at 6 months. The longer, more intensive Vanderbilt University Medical Center program was significantly more effective at helping the civilian portion of the population (85% versus 60% in the American Cancer Society program) but not the active duty participants. PMID- 9358716 TI - Medical support for Operation Cooperative Nugget '95: joint readiness training in the post-cold war era. AB - The purpose of this paper is to report the demographic characteristics, injury and illness profiles, and dispositions of patients seen at the 249th General Hospital during its month-long deployment in support of Operation Cooperative Nugget '95 at the Joint Readiness Training Center (JRTC), Fort Polk, Louisiana. A descriptive analysis of patient demographic, diagnostic, and disposition data was performed. A total of 769 patient contacts were made, with orthopedic injuries (31%), dermatologic disorders (17%), upper respiratory infections (6%), and heat injuries (5%) accounting for the majority of visits. Because of aggressive preventive medicine interventions, there were no cases of heat stroke despite daily heat indices of 110 to 120 degrees F. In addition to emphasizing the importance of anticipating environmental medical threats, the authors relate some lessons learned, which should be valuable to medical providers tasked for future multinational operations other than war at the JRTC and elsewhere. PMID- 9358717 TI - Improved colorectal cancer survival in an army community hospital. AB - In the later portion of 1988, surgeons at Darnall Army Community Hospital abruptly liberalized the use of colonoscopy in the evaluation of colorectal symptoms. During this period, colonoscopy rates jumped from a median of 31.6 examinations/100,000/year to 217.3 examinations/100,000/year. This study details the changes in outcome from colorectal cancer that followed this change. Before 1989, 74.5% of colorectal cancer patients presented with stage III or IV disease, and median survival was only 25 months. After 1988, the proportion of early cancers (stage 0, I, and II) increased dramatically to 56.4% (p = 0.002). Five year survival increased concomitantly from 34.0 to 53.5% (p = 0.042). The liberalization of colonoscopy is judged to have had a beneficial effect on the health of the population served by this community hospital. PMID- 9358718 TI - Family practice house officer perceptions of faculty teaching behaviors. AB - In 1985, Wolverton and Bosworth published an often referenced study of family practice teaching behaviors. Their research identified 20 "most helpful" and 10 "least helpful" behaviors. In 1994 all United States Army family practice house officers were asked by survey to rate each of the original 38 Wolverton and Bosworth teaching behaviors as "not at all helpful," "somewhat helpful," "moderately helpful," "very helpful," or "most helpful" in aiding their learning. Mean values were calculated for each teaching behavior. Twenty most helpful and 10 least helpful faculty teaching behaviors were identified for U.S. Army family practice house officers; several differences were identified in comparison with Wolverton and Bosworth's original study. Additionally, three most helpful and four least helpful teaching behaviors were clearly illuminated. Incorporating the results of this new study into residency programs could improve the learning opportunities afforded family practice house officers during their internships and residencies. PMID- 9358719 TI - Review of the idiopathic bone cavity of the jaws. AB - The IBC is a lesion that favors the anterior portions of the jaws and occurs principally in young patients. The lesion radiographically tends to scallop up between the roots of the affected teeth and result in cortical expansion. The cause of the IBC has not been determined, but trauma seems least likely as the cause. The IBC is best treated by curettage, and follow-up is mandated. A biopsy is required to establish a definitive diagnosis and to rule out aggressive lesions that can appear radiographically similar to the IBC, such as the ameloblastoma and the parakeratinizing odontogenic keratocyst. PMID- 9358720 TI - The psychological status of U.S. Army soldiers during recent military operations. AB - This study compared general psychological symptoms measures on all Brief Symptom Inventory symptom dimensions and the Global Severity Index from samples of deployed and nondeployed U.S. Army soldiers. Psychological symptom measures were taken from samples of soldiers during deployment to operations in the Persian Gulf, Somalia, Kuwait, Haiti, and Bosnia. The purpose of this study was to determine whether deployment and gender had an effect on levels of symptom measures. Results indicated that soldiers who deployed to the Persian Gulf, Somalia, and Bosnia had significantly elevated measures of general psychological distress compared with nondeployed soldiers. Gender difference had little to no effect on reported symptom measures among deployed soldier samples. All female soldiers, whether deployed or not, had elevated measures of interpersonal sensitivity and somatization symptoms. Further research is warranted to address which factors, to include yet not be limited to mission, life events-related, and physical symptoms, may relate to why some deployments are more stressful than others on Army soldiers. PMID- 9358721 TI - A behavioral analysis of eye protection use by soldiers. AB - One of the major problems faced by eye injury prevention programs in the military is the low compliance among individual soldiers with eye armor use. We use three different health behavioral models (the health belief model, the social learning theory, and the PRECEDE model) to analyze and explore the various factors involved in the use of eye armor. Some of the factors that appear to be important in affecting the behavior include environmental conditions (e.g., actual military deployment versus nondeployment activity), organizational attitude toward eye protection programs, community influence, individual knowledge and perception of eye injury, and belief in the efficacy of eye armor. An understanding of these factors can help influence the development of more effective strategies for eye injury prevention in the military. PMID- 9358722 TI - Contrast radiography in small bowel obstruction: a prospective, randomized trial. AB - Small bowel obstruction is a common cause of acute abdominal distress, accounting for up to 20% of emergency admissions to surgical services. Although the majority of obstructions will resolve with conservative therapy alone, there are currently no reliable tests for identifying the patients who will require operation. Barium contrast studies have the potential to rapidly identify patients with complete small bowel obstruction, but many surgeons are hesitant to use them for fear of inducing complications. We report the results of a randomized, prospective trial comparing immediate oral barium contrast studies with plain abdominal X-rays in patients presenting with signs and symptoms of small bowel obstruction. End points included time to resolution of the symptoms or operation, total number of hospital days, and morbidity. Sixty-four patients completed the study; of these, 23 received contrast studies and 41 had plain radiographs only. Six of the contrast group (26%) and 11 of the plain radiograph group (27%) ultimately went to operation. Barium contrast studies had a sensitivity of 100% for diagnosing complete obstruction, whereas the sensitivity of serial plain radiographs was only 82%. Among those going to operation, the time from admission to operation was 8.2 hours in the contrast group and 12.4 hours in the plain radiograph group, but this result did not reach statistical significance (p = 0.25). Total hospital days were similar between the two groups (8 vs. 12 days, p = 0.40). There were no complications resulting from the oral administration of barium. Small bowel contrast studies using barium are safe and may shorten the time to operation in patients presenting with signs and symptoms of small bowel obstruction. PMID- 9358723 TI - Prevalence and contributing factors of eating disorder behaviors in active duty Navy men. AB - Eating disorders have been widely studied among civilian women and among select groups of men (athletes and wrestlers). Gross disturbances in eating behaviors characterize the conditions of anorexia nervosa (AN), currently seen among 1 to 2% of non-active duty women. Bulimia nervosa (BN) is prevalent among 2% of the female population, and both disorders have a female-to-male ratio of 10:1. Another category of eating disorders known as not otherwise specified (NOS) occurs in between 3 and 35% of the population according to the reported literature. This study examined the prevalence of AN, BN, and NOS among a large sample of active duty Navy men. Multiple military, professional, and behavioral were analyzed to promote an increased understanding and awareness of these disorders among active duty men. This was a descriptive, correlational study among 4,800 Navy men targeted from hospitals, clinics, and ships at sea. Anonymous surveys were returned by 28% of prospective subjects (N = 1,425). The study revealed a prevalence of 2.5% for AN, 6.8% for BN, and 40.8% for NOS among active duty Navy men. AN and BN existed without difference among all active duty men regardless of rank/rate, job assignment, or age. Use of laxatives, diuretics, diet pills, vomiting, and fasting for standards increased during the body measurement and fitness periods, but year-round use of these methods existed at disturbing rates. Multiple logistic regression analysis predicted several factors that might predict the manifestation of eating disorders in this population of active duty members. PMID- 9358724 TI - The effectiveness of pressure-reducing table pads as an intervention to reduce the risk of intraoperatively acquired pressure sores. AB - The purpose of this study was to determine the effectiveness of specialty pads as an intervention to reduce the incidence of intraoperatively acquired pressure sores. A convenience sample (N = 361) was drawn from all inpatients who underwent cardiothoracic or major vascular surgery on the standard operating room table (group 1), the air-filled pad (group 2), or the specialty foam pad (group 3). This sample was inclusive of 100% of patients during the study period who met the criteria. The incidence of pressure sore development was seven in group 1, zero in group 2, and one in group 3. There was at statistically significant difference (p = 0.0003) between group 1 and group 2. Additionally, a statistically significant difference (p = 0.0003) was found between group 1 and group 3. The foam pad and the air-filled pad were effective interventions for reducing the risk of intraoperatively acquired pressure sores. PMID- 9358725 TI - Sevoflurane and the 885A field anesthesia machine: clinical report. AB - The Ohmeda 885A field anesthesia machine is equipped with a non-agent-specific, universal vaporizer that can be used with most volatile anesthetic agents. On a recent humanitarian medical mission to Honduras, the 885A was used to administer general anesthesia to 26 patients utilizing sevoflurane, a new inhalational anesthetic with a variety of clinical benefits, including less airway irritability, making it ideal for inhalation inductions. Flow rates for delivery of anesthetic agent were calculated by using the enflurane portion of the Verni Trol flow calculator wheel, because sevoflurane and enflurane have similar vapor pressures. Calculated anesthetic concentrations were compared with measured concentrations using linear regression analysis and found to have a Pearson product moment of 0.995. We find that the use of sevoflurane in the 885A is an excellent alternative to other inhalational anesthetic agents and may have applications for use during both military conflicts and peacetime missions in remote areas. PMID- 9358726 TI - Frequency of infected aneurysms among patients in Department of Veterans Affairs hospitals, 1986-1990: the role of Salmonella. AB - Patients with infected aneurysms discharged from Department of Veterans Affairs hospitals during fiscal years 1986 to 1990 were identified using International Classification of Diseases codes to determine the proportion of patients infected with Salmonella. Twenty-three patients with infected aneurysms were identified. All patients were males; the median age was 63 years. A Gram-positive organism was recovered from 16 patients (70%), a Gram-negative organism from 6 patients (26%), and a fungus from 1 patient. Three patients, all with aneurysms of the abdominal aorta, were infected with Salmonella. Overall, the vessels most often involved included arteries of the extremities (10 patients) and the abdominal or thoracic aorta (9 patients). An increased incidence of nontyphoidal salmonellosis in the United States during the last few decades, along with an aging of the U.S. population, suggests that Salmonella can be expected to maintain a niche in the bacteriology of infected aneurysms. PMID- 9358727 TI - Primary meningococcal arthritis: case report and review of the literature. AB - Meningococcal arthritis is a recognized manifestation of Neisseria meningitidis infection, the presentation of which may be confusing. Although arthritis occurs in the setting of meningococcal meningitis, it may also be seen as a primary event without neurological involvement and with or without cutaneous manifestations. We describe a patient with primary meningococcal arthritis and review the literature relating to the clinical types and pathogenic mechanisms. Comparisons of patient series from 1980 to the present with those reported before 1980 are described. PMID- 9358728 TI - Fabry's disease presenting as syncope, angiokeratomas, and spoke-like cataracts in a young man: discussion of the differential diagnosis. AB - A 44-year-old male with a remote history of skin lesions and cataracts presented with subacute onset of syncope. The presentation and the differential diagnosis in this case of Fabry's disease are discussed. The pathophysiology, diagnosis, clinical manifestations, and treatment of this disorder are discussed, with a review of the literature. Fludrocortisone acetate therapy resulted in a reduction in the patient's syncopal event frequency. Serum enzymatic assay and skin biopsy are useful in the diagnosis of Fabry's disease. We advocate a conservative approach to the oculocutaneous symptoms and suggest appropriate interventions for the renovascular and autonomic nervous system complications of this disorder. PMID- 9358729 TI - Vancomycin: interactions with a model cell membrane. PMID- 9358730 TI - The X-ray structure of the monoclinic crystal form of [D-Hyi2, L-Hyi4] meso valinomycin. AB - The conformation and intermolecular association of [D-Hyi2, L-Hyi4] meso valinomycin [cyclo[-D-Val-D-Hyi-L-Val-L-Hyi-(D-Val-L-Hyi-L-Val-D-+ ++Hyi)2-], C60H102N6O18] in a crystal form obtained from ethanol solution has been determined by x-ray crystallography. Two depsipeptides and one ethanol molecule per asymmetric unit crystallize in space group P2(1) (Z = 4); a = 14.579, b = 39.795, c = 13.928 A, beta = 116.90, Rl = 0.0757. The molecular conformation is very similar to that observed in the trigonal P3(2) crystal form obtained from acetone solution [V. Z. Pletnev et al. (1991) Biopolymers, Vol. 31, pp. 409-415]. Both independent molecules in the crystal adopt a similar distorted bracelet structure with a sterically inaccessible, partially formed, ion-binding center that is stabilized by six 4-->1 type H bonds. The observed conformation accounts for the inability of the molecule to complex ions. Close examination of the three crystallographically independent molecules reveals that differences in the backbone conformation associated with solvent interaction are significantly larger than those associated with hydrophobic van der Waals interactions of crystal packing. PMID- 9358731 TI - The crystal and molecular structure of a valinomycin analogue cyclo[(D-Val-L-Lac L-Ala-D-Hyi)2(D-Val-L-Lac-L-Val-D-Hyi)]. H2O(C50H82N6 O18.H2O). AB - The crystal and molecular structure of the valinomycin analogue, cyclo[(D-Val-L Lac-L-Ala-D-Hyi)2(D-Val-L-Lac-L-Val-D-Hyi)] has been solved by x-ray direct methods using the "Shake and Bake" procedure. The crystals, grown from a mixture of octane/CH2Cl2, belong to space group P2(1) (Z = 4) with cell parameters a = 10.29, b = 32.08, c = 18.73 A, beta = 97.05 degrees, and contain two molecules per asymmetric unit. After anisotropic refinement the standard reliability factor was Rl = 0.058. The conformations of both independent molecules is similar to that observed for isoleucinomycin, cyclo[-(D-Ile-L-Lac-L-Ile-D-Hyi)3] [V. Z. Pletnev et al. (1980) Biopolymers, Vol. 19, pp. 1517-1534]. The structure has an asymmetric conformation stabilized by six intramolecular H bonds, five bonds being of the 4-->1 type and one bond being of the 5-->1 type. One water molecule is caged in the internal cavity of each cyclodepsipeptide. This conformation could represent an intermediate state between free and complexed forms of valinomycin. PMID- 9358732 TI - Oligomerization of smooth muscle myosin light chain kinase and its modifications by melittin and calmodulin. AB - The catalytic activity of smooth muscle myosin light chain kinase (MLCKase) requires the presence of calcium and calmodulin [CM; J. T. Stull et al. (1993) Molecular and Cellular Biochemistry, Vols. 127/128, pp. 229-237] and can also be modified through its own oligomerization [E. B. Babiychuk et al. (1995) Biochemistry, Vol. 34, pp. 6366-6372]. In the present report we demonstrate that melittin, one of the most potent CM antagonists, interacted reversibly with the MLCKase apoenzyme with affinities comparable to those of CM and influenced the oligomeric state of the kinase. At low melittin to kinase ratios the kinase formed insoluble oligomers (aggregates) while at higher melittin concentrations it existed predominantly as soluble oligomers revealed by cross-linking as octamers and hexamers. The kinase alone exhibited similar biphasic solubility within a 5-30 microM range and its solubility was strongly influenced by the ionic strength of the medium. Melittin was also found to promote both the aggregation of the purified 24-kDa C-terminal fragment of the kinase and its analogue telokin, as well as of myosin light chains, but had no effect on the solubility of bovine serum albumin, caldesmon, or calmodulin. These data and our cross-linkage experiments indicate that the insoluble kinase oligomers arose via melittin-induced aggregation of the kinase dimers in which the relokin-like domain played a main role. The soluble oligomers, in turn, were formed after saturation of the kinase with melittin, which resulted in a weakening of the interaction between the protomers with an increase of the long-range order within the oligomers. This interpretation is consistent with the observed effects of melittin on MLCKase catalytic and autocatalytic activities. At concentrations of melittin required to produce soluble oligomers, the binding of the kinase to myosin filaments was considerably enhanced. A plausible mechanism for the formation of the soluble oligomers and aggregates is suggested and its relation to the possible MLCKase assemblies discussed in terms of a model. PMID- 9358733 TI - The free solution mobility of DNA. AB - The free solution mobility of DNA has been measured by capillary electrophoresis in the two buffers most commonly used for DNA gel electrophoresis, Tris-borate EDTA (TBE) and Tris-acetate-EDTA (TAE). The capillaries were coated with polymers of either of two novel acrylamide monomers, N-acryloylaminoethoxyethanol or N acryloylaminopropanol, both of which are stable at basic pH and effectively eliminate the electroendosmotic mobility due to the capillary walls. The free solution mobility of DNA in TAE buffer was found to be (3.75 +/- 0.04) x 10(-4) cm2 V-1 s-1 at 25 degrees C, independent of DNA concentration, sample size, electric field strength, and capillary coating, and in good agreement with other values in the literature. The free solution mobility was independent of DNA molecular weight from approximately 400 base pairs to 48.5 kilobase pairs, but decreased monotonically with decreasing molecular weight for smaller fragments. Surprisingly, the free solution mobility of DNA in TBE buffer was found to be (4.5 +/- 0.1) x 10(-4) cm2 V-1 s-1, about 20% larger than observed in TAE buffer, presumably because of the formation of nonspecific borate-deoxyribose complexes. PMID- 9358734 TI - Model for the alignment of actin filaments in endothelial cells subjected to fluid shear stress. AB - Cultured vascular endothelial cells undergo significant morphological changes when subjected to sustained fluid shear stress. The cells elongate and align in the direction of applied flow. Accompanying this shape change is a reorganization at the intracellular level. The cytoskeletal actin filaments reorient in the direction of the cells' long axis. How this external stimulus is transmitted to the endothelial cytoskeleton still remains unclear. In this article, we present a theoretical model accounting for the cytoskeletal reorganization under the influence of fluid shear stress. We develop a system of integro-partial differential equations describing the dynamics of actin filaments, the actin binding proteins, and the drift of transmembrane proteins due to the fluid shear forces applied on the plasma membrane. Numerical simulations of the equations show that under certain conditions, initially randomly oriented cytoskeletal actin filaments reorient in structures parallel to the externally applied fluid shear forces. Thus, the model suggests a mechanism by which shear forces acting on the cell membrane can be transmitted to the entire cytoskeleton via molecular interactions alone. PMID- 9358735 TI - Quantal currents and potential in the three-dimensional anisotropic bidomain model of smooth muscle. AB - The potential generated in the smooth muscle of the vas deferens on release of a quantum of transmitter from a varicosity was analyzed using a three-dimensional bidomain continuum model. Current was injected at the origin of the bidomain; this current had the temporal characteristics of the junctional current. The membrane potential, intracellular potential, and extracellular potential, as well as the extracellular current, were then calculated throughout the bidomain at different times. Calculations were performed to show the effect of changing the anisotropy ratios of the intracellular and extracellular conductivities on the spread of current and potential in each of the three dimensions. These results provide a theoretical framework for ascertaining the time course of transmitter interaction at a varicosity following the secretion of a quantum of transmitter. PMID- 9358737 TI - Agonist-induced calcium waves in oscillatory cells: a biological example of Burgers' equation. AB - Oscillations in cytosolic Ca2+ concentrations in living cells are often a manifestation of propagating waves of Ca2+. Numerical simulations with a realistic model of inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ wave trains lead to wave speeds that increase linearly at long times when (a) IP3 levels are in the range for Ca2+ oscillations, (b) a gradient of phase is established by either an initial ramp or pulse of IP3, and (c) IP3 concentrations asymptotically become uniform. We explore this phenomenon with analytical and numerical methods using a simple two-variable reduction of the De Young-Keizer model of the IP3 receptor that includes the influence of Ca2+ buffers. For concentrations of IP3 in the oscillatory regime, numerical solution of the resulting reaction diffusion equations produces nonlinear wave trains that shows the same asymptotic growth of wave speed. Due to buffering, diffusion of Ca2+ is quite slow and, as previously noted, these waves occur without appreciable bulk movement of Ca2+. Thus, following Neu and Murray, we explore the behavior of these waves using an asymptotic expansion based on the small size of the buffered diffusion constant for Ca2+. We find that the gradient in phase of the wave obeys Burgers' equation asymptotically in time. This result is used to explain the linear increase of the wave speed observed in the simulations. PMID- 9358738 TI - Qualitative modeling of mechanoelectrical feedback in a ventricular cell. AB - Mechanical changes in the heart muscle can influence its electrical properties through a process called mechanoelectrical feedback (MEF). This feedback can operate via changes in calcium dynamics during the cross-bridge cycle or via mechanosensitive (stretch-activated) channels. We present a four-variable ordinary differential equation (ODE) system that caricatures the electrical and mechanical activity of a ventricular cell and their mutual interactions. A three variable excitable system with restitution properties of the FitzHugh-Nagumo type is coupled to a fourth equation which describes changes in cell length during a lightly loaded contraction. The resulting four-variable system models MEF in a cell and can be incorporated into spatially distributed models for mechanoelectric behavior during wave propagation in the cardiac tissue. PMID- 9358736 TI - Role of fibroblast migration in collagen fiber formation during fetal and adult dermal wound healing. AB - Adult dermal wounds, in contrast to fetal wounds, heal with the formation of scar tissue. A crucial factor in determining the degree of scarring is the ratio of types I and III collagen, which regulates the diameter of the combined fibers. We developed a reaction-diffusion model which focuses on the control of collagen synthesis by different isoforms of the polypeptide transforming growth factor beta (TGF beta). We used the model to investigate the current controversy as to whether the fibroblasts migrate into the wound from the surrounding unwounded dermis or from the underlying subcutaneous tissue. Numerical simulations of a spatially independent, temporal model led to a value of the collagen ratio consistent with that of healthy tissue for the fetus, but corresponding to scarring in the adult. We investigated the effect of topical application of TGF beta and show that addition of isoform 3 reduces scar tissue formation, in agreement with the experiment. However, numerical solutions of the reaction diffusion system do not exhibit this sensitivity to growth factor application. Mathematically, this corresponds to the observation that behind healing wave front solutions, a particular healed state is always selected independent of transients, even though there is a continuum of possible positive steady states. We explain this phenomenon using a caricature system of equations, which reflects the key qualitative features of the full model but has a much simpler mathematical form. Biologically, our results suggest that the migration into a wound of fibroblasts and TGF beta from the surrounding dermis alone cannot account for the essential features of the healing process, and that fibroblasts entering from the underlying subcutaneous tissue are crucial to the healing process. PMID- 9358739 TI - Cell communities and robustness in development. AB - The robustness of patterning events in development is a key feature that must be accounted for in proposed models of these events. When considering explicitly cellular systems, robustness can be exhibited at different levels of organization. Consideration of two widespread patterning mechanisms suggests that robustness at the level of cell communities can result from variable development at the level of individual cells; models of these mechanisms show how interactions between participating cells guarantee community-level robustness. Cooperative interactions enhance homogeneity within communities of like cells and the sharpness of boundaries between communities of distinct cells, while competitive interactions amplify small inhomogeneities within communities of initially equivalent cells, resulting in fine-grained patterns of cell specialization. PMID- 9358740 TI - What mediates progressive glomerulosclerosis? The glomerular endothelium comes of age. PMID- 9358741 TI - Profound effects on vascular development caused by perturbations during organogenesis. PMID- 9358742 TI - Hepatocytes break the rules of senescence in serial transplantation studies. Is there a limit to their replicative capacity? PMID- 9358743 TI - The integrin alpha 6 beta 1 promotes the survival of metastatic human breast carcinoma cells in mice. AB - The role of the integrin alpha 6 beta 1 in breast carcinoma progression was studied by targeted elimination of this integrin in MDA-MB-435 cells, a human breast carcinoma cell line that is highly metastatic in athymic mice. The strategy used is based on the finding that expression of a cytoplasmic domain deletion mutant of the beta 4-integrin subunit (beta 4-delta CYT) in MDA-MB-435 cells eliminates formation of the alpha 6 beta 1 heterodimer. MDA-MB-435 cells that lacked alpha 6 beta 1 expression (beta 4-delta CYT transfectants) formed tumors in athymic mice that were suppressed in their growth and that exhibited a significant increase in apoptosis in comparison to the control tumors. Unlike the control MDA-MB-435 cells, the beta 4-delta CYT transfectants were unable to establish metastatic foci in the lungs. Also, the control transfectants grew substantially better than the beta 4-delta CYT transfectants in the liver after intrahepatic injection because of extensive apoptosis in the beta 4-delta CYT transfectants. These data suggest that a major function of the alpha 6 beta 1 integrin in breast carcinoma is to facilitate tumorigenesis and promote tumor cell survival in distant organs. PMID- 9358744 TI - Mice lacking the thrombin receptor, PAR1, have normal skin wound healing. AB - Thrombin's actions on platelets, macrophages, fibroblasts, and endothelial cells have prompted the hypothesis that thrombin may be important for inflammatory and fibroproliferative processes in wound healing. Protease-activated receptor 1 (PAR1) is a G-protein-coupled receptor that mediates many of the cellular activities of thrombin. To test the role of this receptor in vivo, we generated PAR1-deficient mice. Despite the observation that fibroblasts cultured from these mice lacked responsiveness to thrombin in vitro, we now report that there was no difference detected between wild-type and PAR1-deficient mice in skin wound healing assays including time to closure of open wounds, tensile strength of healed incisional wounds, wound histology, and hydroxyproline/DNA content of wound implants. We conclude that PAR1 is not necessary for normal skin wound healing in mice. PMID- 9358746 TI - Systemic activation of the vascular endothelial growth factor receptor KDR/flk-1 selectively triggers endothelial cells with an angiogenic phenotype. AB - The hypothesis that tumor growth is angiogenesis dependent has been documented by a considerable body of direct and indirect experimental data. A prerequisite for the development of novel anti-angiogenic agents is the design of drugs that would be active only on those endothelial cells with an angiogenic phenotype. We took advantage of the anti-idiotypic strategy to obtain circulating agonists specific for the vascular endothelial growth factor receptor KDR/flk-1 (J-IgG). They induced in the absence of VEGF cell proliferation in vitro and angiogenesis in the corneal pocket assay either through local or systemic delivery. Intraperitoneal injections of J-IgG in nude mice grafted with a prostatic adenocarcinoma led to tumor enlargement associated with an increase in both tumor vascularization and proliferation. In contrast KDR/flk-1 overstimulation had no detectable effect on normal tissues. These data underline that KDR/flk-1 is a functional marker of the angiogenic phenotype of endothelial cells. PMID- 9358745 TI - Internucleosomal DNA cleavage triggered by plasma membrane damage during necrotic cell death. Involvement of serine but not cysteine proteases. AB - Autolytic DNA breakdown, detected as smears in electrophoretic gels, is a late event in necrosis. On the other hand, internucleosomal DNA cleavage, visualized as ladders, is thought to be a hallmark of apoptosis. We now report that this specific form of DNA fragmentation also occurs during necrosis and is an early event but appears to be triggered by proteolytic mechanisms significantly different from those documented in apoptosis. Treatment of MDCK cells with a mitochondrial uncoupler and a Ca2+ ionophore led to ATP depletion, necrotic morphology, and progressive fragmentation of DNA in an internucleosomal or ladder pattern. DNA breakdown was immediately preceded by increased permeability of the plasma membrane to macromolecules. Provision of glycine along with the noxious agents did not modify the extent of ATP depletion, but prevented plasma membrane damage. This was accompanied by complete inhibition of DNA fragmentation. Internucleosomal DNA cleavage was observed also during necrosis after rapid permeabilization of plasma membranes by detergents or streptolysin-O in hepatocytes, thymocytes, and P19, Jurkat, and MDCK cells. DNA fragmentation associated with necrosis was Ca2+/Mg2+ dependent, was suppressed by endonuclease inhibitors, and was abolished by serine protease inhibitors but not by inhibitors of interleukin-1 beta converting enzyme (ICE)-related proteases or caspases. Moreover, unlike apoptosis, it was not accompanied by caspase-mediated proteolysis. On the other hand, the cleavage-site-directed chymotryptic inhibitor N-tosyl-L-phenylalanyl-chloromethyl ketone (TPCK) suppressed DNA fragmentation not only in necrotic cells but also during Fas-mediated apoptosis, without inhibiting caspase-related proteolysis. The results suggest a novel pathway of endonuclease activation during necrosis not involving the participation of caspases. In addition, they indicate that techniques based on double-strand DNA breaks may not reliably differentiate between apoptosis and necrosis. PMID- 9358747 TI - N-acetyl cysteine blocks mesangial VCAM-1 and NF-kappa B expression in vivo. AB - Inducible vascular cell adhesion molecule-1 (VCAM-1) in glomerular mesangial cells (GMC) exposed to lipopolysaccharide (LPS) in vitro involves the activation of nuclear factor-kappa B (NF-kappa B) and its interaction with the proximal VCAM 1 promoter. We used a murine model to assess the effect of the antioxidant, N acetyl cysteine on GMC activation in vivo. Single intraperitoneal administration of N-acetyl cysteine completely suppressed LPS-induced VCAM-1 expression on the GMC surface. When an oligonucleotide spanning the NF-kappa B binding region of the VCAM-1 promoter was incubated with extracts from the renal cortex of LPS treated animals, a single nucleoprotein complex formed. This complex was composed of p50 and p65, but not p52, c-Rel, or RelB, and its formation was dramatically inhibited by pretreatment with N-acetyl cysteine, D,L-Buthionine-[S,R] sulfoximide, a compound that depletes glutathione, augmented VCAM-1 expression inducible with a suboptimal amount of LPS to levels comparable with using 50 micrograms of LPS alone, D,L-Buthionine-[S,R]-sulfoximide also potentiated the p50-p65 binding activity induced with a suboptimal amount of LPS. These data provide a redox-sensitive, transcriptional link between NF-kappa B and VCAM-1 in GMC in vivo and implicate oxidative stress as an important regulatory signal in the pathogenesis of glomerular mesangial cell disorders. PMID- 9358748 TI - Rare glomerular capillary regeneration and subsequent capillary regression with endothelial cell apoptosis in progressive glomerulonephritis. AB - Glomerulonephritis (GN) leading to glomerular sclerosis remains an important cause of renal failure. The glomerulus is a capillary network, but endothelial and vascular reactions during progressive GN are not well understood. We have, therefore, examined the morphological alterations of glomerular capillary network and endothelial cells during the progression of damaged glomeruli to glomerular sclerosis. A progressive model of anti-glomerular basement membrane (GBM) GN was induced in Wistar-Kyoto (WKY) rats with a single injection of anti-rat GBM antibody. Severe necrotizing glomerular injuries were observed between day 5 and week 3 with a reduction in the number of total glomerular endothelial cells and total glomerular capillary lumina per glomerular cross sections. In necrotizing lesions, the glomerular endothelial cells were lost with the destruction of the glomerular capillary network. Moreover, angiogenic capillary repair with proliferation of endothelial cells was rare in severely damaged regions of glomeruli. Subsequently, mesangial hypercellularity and marked mesangial matrix accumulation occurred with absence of the development of a capillary network, and the necrotizing lesions progressed to sclerotic scars until 8 weeks. Although active necrotizing lesions could not be seen in damaged glomeruli between week 4 and week 8, the number of apoptotic endothelial cells gradually increased in the glomerular capillaries (0.10 +/- 0.01 apoptotic endothelial cells/glomerular cross section at week 8 versus 0.00 +/- 0.00 control cells (mean +/- SEM; P < 0.05) with the progression of glomerular sclerosis. Whereas the number of apoptotic endothelial cells increased in the damaged glomeruli, the number of total glomerular endothelial cells decreased (9.3 +/- 3.0 cells/glomerular cross section at week 8 versus 24.8 +/- 3.0 cells in control (mean +/- SD); P < 0.001) with regression of glomerular capillaries (3.6 +/- 2.5 capillary lumina/glomerular cross section at week 8 versus 35.0 +/- 5.0 capillary lumina in control (mean +/- SD); P < 0.001). Finally, glomerular endothelial cells could not be detected in the sclerotic lesions in progressive anti-GBM GN in WKY rats. These data indicate that the destruction of the capillary network of glomeruli and subsequent incomplete angiogenic capillary repair leads to glomerular sclerosis in progressive GN. Endothelial cell apoptosis with glomerular capillary regression may also contribute to the development of glomerular sclerosis. Injury of the glomerular capillary network with endothelial cell damage, including apoptosis and subsequent incomplete capillary repair, plays an important role in the progression of glomerular sclerosis during anti-GBM GN in WKY rats. PMID- 9358749 TI - Time course and localization of endothelin-1 gene expression in a model of renal disease progression. AB - Experimental and human proteinuric glomerulopathies are associated with tubulo interstitial injury that correlates with the decline of renal function even better than glomerular lesions do. Mechanism(s) leading to tubulo-interstitial damage are unknown. It has been proposed that excessive reabsorption of filtered proteins activates renal cells to produce vasoactive and inflammatory molecules including endothelin-1. The aim of the present study was twofold: we first evaluated the cellular origin of excessive renal endothelin-1 production in the renal mass reduction model and then related endothelin-1 distribution to the development of kidney lesions. Four groups of renal mass reduction (n = 15) and four groups of control rats (n = 5) were studied at 7, 14, 21, and 28 days after surgery. Urinary proteins in renal mass reduction rats were comparable with controls at day 7 but became significantly higher thereafter. Renal mass reduction rats first developed tubulo-interstitial changes, which were already evident at day 14 in the majority of them. At 28 days, renal mass reduction rats also developed glomerulosclerosis. A parallel increase of renal endothelin-1 gene expression and synthesis of the corresponding peptide in renal mass reduction rats versus controls was observed from day 14. Nonradioactive in situ hybridization confirmed a pattern of endothelin-1 mRNA consistent with the distribution of lesions. At day 14, endothelin-1 staining was stronger in renal mass reduction than in control kidneys and mainly localized to the cytoplasm of tubular cells, whereas glomeruli were negative. At day 28, endothelin-1 expression further increased in renal mass reduction rats as compared with controls, and the staining was apparent also in glomeruli. Thus, in renal mass reduction, a progressive up-regulation of endothelin-1 occurs during the development of renal injury, that first involves the tubules and, only in a subsequent phase, the glomeruli. PMID- 9358750 TI - Intraglomerular C3 synthesis in rats with passive Heymann nephritis. AB - Passive Heymann nephritis (PHN), a model of human membranous nephropathy, is an immune-complex-mediated glomerulonephritis characterized by the presence of complement-dependent tissue injury. Recent studies have confirmed the synthesis of C3, involved in both the classical and alternative pathways of complement, in injured human and animal renal tissues. However, there is little clear information on the role of local C3 synthesis in the pathogenesis of nephritides such as PHN. In the present study, using nonradioactive in situ hybridization and semiquantitative reverse transcriptase polymerase chain reaction, we examined C3 synthesis in the kidney and its contribution to tissue injury in a rat model of PHN induced by the injection of polyclonal anti-gp330 antibody. C3 mRNA was localized in mesangial cells, glomerular epithelial cells, and cells of Bowman's capsule. During the early stages of PHN, C3 mRNA expression was detected in mesangial cells and glomerular epithelial cells, whereas such expression was limited to mesangial cells during the late stages of the disease. Focal, weak C3 mRNA expression was detected in tubular epithelial cells and occasionally in the interstitium. Semiquantitative polymerase chain reaction demonstrated that the level of C3 mRNA expression correlated with that of proteinuria. Our results suggest that renal cells synthesize C3 mRNA in PHN in a site-specific manner and that locally produced C3 is associated with the development of proteinuria in this model. PMID- 9358751 TI - Ceramide is involved in triggering of cardiomyocyte apoptosis induced by ischemia and reperfusion. AB - Involvement of ceramide signaling in the initiation of apoptosis induction in myocardial cells by in vitro and in vivo ischemia and reperfusion was analyzed. Synthetic cell permeable C2-ceramide induced apoptotic death of rat neonatal cardiomyocytes in vitro. In vitro ischemia (oxygen/serum/glucose deprivation) led to a progressive accumulation of ceramide in cardiomyocytes. After 16 hours of simulated in vitro reperfusion (readdition of oxygen, serum and glucose), the level of ceramide in surviving cells was found to have returned to baseline, whereas, levels in nonadherent dead cells remained high. In the rat heart left coronary artery occlusion model, ischemia with the subsequent reperfusion, but not ischemia alone, induced apoptosis in myocardial cells as demonstrated by DNA electrophoresis and measurement of soluble chromatin degradation products. The content of ceramide in ischemic area was elevated to 155% baseline levels at 30 minutes, and to 330% after 210 minutes of ischemia. Ischemia (30 minutes) followed by reperfusion (180 minutes) increased the ceramide level to 250% in the ischemic area. The combination of results obtained in both in vitro and animal models demonstrate for the first time that ceramide signaling can be involved in ischemia/reperfusion death of myocardial cells. PMID- 9358752 TI - CD95/CD95L-mediated apoptosis of the hepatic stellate cell. A mechanism terminating uncontrolled hepatic stellate cell proliferation during hepatic tissue repair. AB - During liver tissue repair, hepatic stellate cells (HSC), a pericyte-like mesenchymal liver cell population, transform from a "quiescent" status ("resting" HSC) into myofibroblast-like cells ("activated" HSC) with the latter representing the principle matrix synthesizing cell of the liver. Presently, the mechanisms that terminate HSC cell proliferation when tissue repair is concluded are poorly understood. Controlled cell death known as apoptosis could be a mechanism underlying this phenomenon. Therefore, apoptosis and its regulation were studied in HSC using an in vitro and in vivo approach. Spontaneous apoptosis became detectable in parallel with HSC activation because resting cells (2 days after isolation) displayed no sign of apoptosis, whereas apoptosis was present in 8% (+/- 5%) of "transitional" cells (day 4) and in 18% (+/- 8%) of fully activated cells (day 7). Both CD95 (APO-1/Fas) and CD95L (APO-1-/Fas-ligand) became increasingly expressed during the course of activation. Apoptosis could be fully blocked by CD95-blocking antibodies in normal cells and HSC already entering the apoptotic cycle. Using CD95-activating antibodies, transition of more than 95% cells into apoptosis was evident at each activation step. The apoptosis regulating proteins Bcl-2 and p53 could not be detected in resting cells but were found in increasing amounts at days 4 and 7 of cultivation. Whereas p53 expression was induced by the CD95-activating antibody, no change was inducible in Bcl-2 expression. The Bcl-2-related protein bax could be found at days 2 and 4 in similar expression, was considerably up-regulated at day 7, but was not regulated by CD95-agonistic antibodies. In vivo, acute tissue damage was first accompanied by activation and proliferation of HSC displaying no sign of apoptosis. In the recovery phase, apoptotic HSC were detectable in parallel to a reduction in the total number of HSC present in the liver tissue. The data demonstrate that apoptosis becomes detectable in parallel with HSC activation, which suggests that apoptosis might represent an important mechanism terminating proliferation of activated HSC. PMID- 9358753 TI - Serial transplantation reveals the stem-cell-like regenerative potential of adult mouse hepatocytes. AB - Previous work has shown that adult mouse hepatocytes can divide at least 18 times in vivo. To test whether this represents the upper limit of their regenerative capacity, we performed serial transplantation of hepatocytes in the fumarylacetoacetate hydrolase deficiency murine model of liver repopulation. Hepatocytes from adult donors were serially transplanted in limiting numbers six times and resulted in complete repopulation during each cycle. This corresponds to a minimal number of 69 cell doublings or a 7.3 x 10(20)-fold expansion. No evidence for abnormal liver function or altered hepatic architecture was found in repopulated animals. We conclude that a fraction of adult mouse hepatocytes have growth potential similar to that of hematopoietic stem cells. PMID- 9358754 TI - Lacrimal gland inflammation is responsible for ocular pathology in TGF-beta 1 null mice. AB - Mice homozygous for a nonfunctional transforming growth factor-beta 1 gene develop rampant inflammation in vital organs that contributes to a shortened life span. The presence of circulating anti-nuclear anti-bodies, immune deposits in tissues, leukocyte infiltration, and increased major histocompatibility complex antigen expression resembles an autoimmune-like syndrome. One of the overt symptoms that appears in these mice lacking transforming growth factor-beta 1 is the development of dry crusty eyes that close persistently as their health declines. Histologically, the eyes appear normal with little or no inflammation. However, inflammatory lesions, predominantly lymphocytic, develop in the lacrimal glands, disrupting their structure and function and severely limiting their ability to generate tears. This histopathology and aberrant function mimic that of Sjogren's syndrome, a human autoimmune disease characterized by dry eyes and dry mouth. Impeding the leukocyte infiltration into the glands with synthetic fibronectin peptides, which block adhesion, not only prevents the inflammatory pathology but also prevents the persistent eye closure characteristic of these mice. PMID- 9358755 TI - Pancreatic carcinomas deposit laminin-5, preferably adhere to laminin-5, and migrate on the newly deposited basement membrane. AB - We studied the adhesion mechanism of pancreatic carcinoma using in vitro adhesion and migration assays of stable cell lines and tumors grown from these cell lines in nude mice. We also compared the results with the expression profiles of laminins and their receptors in pancreatic carcinomas to evaluate the relevance of these mechanisms in vivo. All of the cell lines preferably adhered to laminin 5, irrespective of their capability to synthesize laminin-5. Cell migration was studied in the presence of hepatocyte growth factor, as it increased the speed of migration manyfold. Herbimycin A treatment and antibodies against the beta 1 and alpha 3 integrin subunits and laminin alpha 3 chain almost entirely blocked cell migration of the BxPC-3 cell line, whereas migration was nearly unaffected by RGD peptide and only moderately inhibited by antibody against the alpha 6 integrin subunit. Indirect immunofluorescence microscopy of wounded BxPC-3 cells suggested a rapid endocytosis of alpha 3 integrin subunit in the cells at the margin of the wound and a rapid, polarized rearrangement of the alpha 6 beta 4 integrin. Especially HGF-treated cultures showed a prominent cytoplasmic reaction for laminin-5 at the margin of the wound. Xenografted cells formed tumors that produced and deposited the same laminin chains as the in vitro cultures. Frozen sections of human pancreatic carcinomas showed reactivity for laminin chains suggestive for expression of laminin-1 and laminin-5. Both xenografted tumors and human pancreatic carcinomas also showed stromal reactivity for laminin-5. Electron microscopy of the human tumors suggested that this was due to an abundant reduplication the basement-membrane-like material around the nests of malignant cells. Our results suggest that pancreatic carcinomas synthesize and deposit laminin-5 in the basement membrane in an abnormal manner. Invading cells adhere to this newly produced basement membrane and migrate on it by using the alpha 3 beta 1 integrin receptor recognizing laminin-5. PMID- 9358756 TI - Role of tumor necrosis factor-alpha in the spontaneous development of pulmonary fibrosis in viable motheaten mutant mice. AB - The viable motheaten mutant mouse is severely immunodeficient and dies from a naturally occurring progressive pulmonary inflammation at approximately 10 weeks of age. The pulmonary disease is characterized by excessive macrophage accumulation in the lung and fibrosis. We correlated the development of lung injury in viable motheaten mice with tumor necrosis factor-alpha (TNF-alpha) levels in serum and lung. Significantly increased serum TNF-alpha levels were observed by enzyme-linked immunosorbent assay in viable motheaten mice at 4, 6, and 10 weeks of age as compared with normal control littermate mice. These ages correlated well with observed changes in lung wet weights, lung collagen content, and histological evidence of pulmonary inflammation and fibrosis. Qualitative assessment of lung tissue TNF-alpha levels was performed by immunohistochemical staining using immunoperoxidase techniques. These studies revealed increased levels of TNF-alpha in macrophage-like cells in viable motheaten mice from 5 to 10 weeks of age as compared with littermate control animals. Alveolar macrophages isolated from viable motheaten mice produced significantly greater amounts of TNF alpha when stimulated with lipopolysaccharide compared with alveolar macrophages from control animals. In addition, administration of anti-TNF-alpha antibody to motheaten bone marrow recipient mice decreased the severity of acute lung injury. The results demonstrate a close correlation between the development of pulmonary injury and TNF-alpha levels in this model of spontaneously developing fibrotic lung disease. PMID- 9358757 TI - Tissue cytokine patterns distinguish variants of rheumatoid synovitis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease with primary manifestations in the synovial membrane. Tissue infiltrates are composed of T cells, B cells, and macrophages, but histopathological appearances vary widely and are rarely pathognomonic. Mechanisms underlying the phenotypic heterogeneity of rheumatoid synovitis are not known. To explore whether a correlation exists between the microscopic patterns of rheumatoid synovitis and in situ production of cytokines, tissue samples from 21 consecutive patients with clinically active RA were examined. Based upon the organization of the lymphocyte infiltrate, the synovial biopsies were categorized into three distinct subsets. Ten samples were characterized by diffuse lymphoid infiltrates without further microarrangement. In seven samples, lymphoid follicles with germinal center formation were detected, and in four specimens, granuloma formation was identified. In all specimens, cytokine transcription of interferon (IFN)-gamma, interleukin (IL)-4, IL-1 beta, tumor necrosis factor (TNF)-alpha, IL-10, and transforming growth factor-beta 1 was semiquantified with polymerase chain reaction and liquid phase hybridization. Each of the morphologically defined variants of synovitis displayed a unique cytokine profile. Low-level transcription of IFN-gamma, IL-4, IL-1 beta, and TNF-alpha was typical of diffuse synovitis. In follicular synovitis, IFN-gamma was the dominant cytokine, IL-4 was virtually undetectable, and IL-10 was abundant. Granulomatous synovitis demonstrated high transcription of IFN-gamma, IL-4, IL-1 beta, and TNF-alpha and could be clearly distinguished from the other phenotypes. To investigate whether differences in the synovial lesions were related to host factors, patients were compared for clinical parameters. Diffuse synovitis was seen in most of the patients with seronegative RA, the mildest form of the disease. In contrast, extra-articular spreading of RA with nodule formation was typically associated with granulomatous synovitis. In summary, RA patients display reproducible patterns in the organization and activity of synovial infiltrates. The correlation of microanatomy with tissue cytokine production suggests that several pathomechanisms can modulate the expression of the immune response in the synovial membrane. PMID- 9358759 TI - Differential expression of the immediate-early and early antigens in neuronal and glial cells of developing mouse brains infected with murine cytomegalovirus. AB - Brain disorders induced by congenital cytomegalovirus (CMV) infection may appear at a later time after birth as a consequence of persistent infection and/or the activation of a latent infection of the neural cells. We have analyzed the infection dynamics of the neural cells in the neonatal mouse brains infected with murine CMV (MCMV) in the late stage of gestation. First we prepared a rat monoclonal antibody to the major immediate-early (IE)-89K antigen and then used the antibody for comparison of the expression of early and late viral genes in the developing mouse brains. The cells expressing the IE-89K antigen were mostly localized in the ventricular and subventricular zones and were preferentially double stained with anti-glial fibrillary acidic protein and anti-nestin antibodies. In contrast, the cells expressing the early nuclear antigen, detected by the monoclonal antibody D5, were diffusely distributed in the cortex and the hippocampus and were mostly double labeled with anti-neuron-specific enolase antibody. In neonatal mouse brains infected congenitally with recombinant MCMV, which expressed lacZ as a late gene, the number of the early nuclear antigen positive cells was much higher than that of the beta-galactosidase-expressing cells, the number of which was almost the same as that of the IE-89K antigen positive cells. In addition, the distribution of viral DNA-rich cells detected by DNA-DNA hybridization was similar to that of the IE-89K antigen-positive cells. These results suggest that CMV may persistently infect neuronal cells, whereas lytic infection may preferentially occur in the glial cells in the developing brain. PMID- 9358758 TI - Keratinocytes derived from psoriatic plaques are resistant to apoptosis compared with normal skin. AB - Previously we observed that hyperplastic epidermal keratinocytes characteristic of psoriasis had abundant amounts of the cell survival protein Bcl-xL; however, whether this overexpression correlated with enhanced survival was unclear because the majority of epidermal cells possess nuclei that are positively labeled by an assay typically regarded as indicative of cells undergoing apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) staining). To clarify this apparent discrepancy, we explored the propensity of keratinocytes derived from psoriatic plaques to undergo apoptosis and also determined the reliability of TUNEL staining as an indicator of apoptosis in keratinocytes in vitro and in vivo. First, a keratinocyte cell line, HaCat, was examined before and after being suspended in semisolid medium (methylcellulose) using flow cytometry to detect TUNEL-positive cells, and the percentage of positive cells was correlated to the presence or absence of double-stranded DNA fragmentation using pulsed field gel electrophoresis. After 18 hours in methylcellulose suspension, apoptosis was detected in HaCat cells when at least 5% of the cell population was undergoing programmed cell death. Second, we examined 23 clinical specimens of skin (13 from psoriatic patients and 10 from healthy control subjects) and observed that no double-stranded DNA fragmentation was present in any of the freshly isolated keratinocytes from either normal or psoriatic patients. Keratinocytes from 9 of 12 normal skin samples underwent double-stranded DNA fragmentation after being in methylcellulose for 18 to 24 hours, which contrasts with keratinocytes from lesions of psoriasis where only 1 of 13 of the skin samples had these changes. Third, two-color immunofluorescence staining of psoriatic plaques revealed that numerous TUNEL-positive keratinocytes were also positive for proliferating cell nuclear antigen and Ki-67 antigens and that by flow cytometry TUNEL-positive keratinocytes obtained from psoriatic plaques possessed a DNA content profile indicative of proliferating and not dying cells. These results demonstrate that keratinocytes within psoriatic plaques do not have double-stranded DNA breaks, that they have a prolonged capacity to resist induction of apoptosis compared with normal-skin-derived keratinocytes or cultured HaCat cells, and that caution is necessary for proper interpretation related to detection of 3'-OH DNA ends (ie, TUNEL positivity) in skin, as it can be associated with DNA synthesis as well as cell death. PMID- 9358760 TI - Cellular localization of the chemokine receptor CCR5. Correlation to cellular targets of HIV-1 infection. AB - The chemokine receptor CCR5 has recently been described as a co-receptor for macrophage-tropic strains of human immunodeficiency virus (HIV)-1. In this study, using a panel of monoclonal antibodies specific for human CCR5, we show by immunohistochemistry and flow cytometry that CCR5 is expressed by bone-marrow derived cells known to be targets for HIV-1 infection, including a subpopulation of lymphocytes and monocyte/macrophages in blood, primary and secondary lymphoid organs, and noninflamed tissues. In the central nervous system, CCR5 is expressed on neurons, astrocytes, and microglia. In other tissues, CCR5 is expressed on epithelium, endothelium, vascular smooth muscle, and fibroblasts. Chronically inflamed tissues contain an increased number of CCR5+ mononuclear cells, and the number of immunoreactive cells is directly associated with a histopathological correlate of inflammatory severity. Collectively, these results suggest that CCR5+ cells are recruited to inflammatory sites and, as such, may facilitate transmission of macrophage-tropic strains of HIV-1. PMID- 9358761 TI - Characterization of tumor-infiltrating T lymphocytes in B-cell lymphomas of mucosa-associated lymphoid tissue. AB - B-cell lymphomas of mucosa-associated lymphoid tissue invariably contain large numbers of reactive tumor-infiltrating T cells. In the stomach, these lymphomas develop secondary to Helicobacter pylori infection, and clinical and in vitro studies have shown that their growth depends on help provided by H. pylori specific T cells. In this study we characterized tumor-infiltrating T cells in low- and high-grade B-cell lymphomas of mucosa-associated lymphoid tissue using immunohistochemistry. In most cases, CD4+ T cells dominated and almost all T cells were CD45RO+ memory cells. In 11 of 13 cases studied, the proliferating T cells were CD4+ and no proliferation was observed in the CD8+ subset. In low grade lymphomas, between 7 and 24% of T cells expressed CD40L whereas no CD40L expression was observed in the majority of high-grade tumors. Examination of homing receptor profile showed that both alpha 4 beta 7 integrin+ and L-selectin+ T cells were present. Examination of T cell diversity by a panel of antibodies against different T-cell receptor V beta regions and by analysis of T-cell receptor genes using polymerase chain reaction suggested that the T cells in these tumors were polyclonal. These results show that low-grade B-cell lymphomas of mucosa-associated lymphoid tissue contain a significant population of activated helper T cells that may be important in supporting tumor growth. PMID- 9358762 TI - Expression of lymphocyte homing receptors and vascular addressins in low-grade gastric B-cell lymphomas of mucosa-associated lymphoid tissue. AB - In this study, we examined lymphocyte homing receptor and vascular addressin expression in a case of primary gastric B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) with a secondary intestinal spread. We compared the findings with that observed in B cells of normal MALT and MALT acquired as a consequence of Helicobacter pylori-associated gastritis and other low-grade gastric B-cell MALT lymphomas. The neoplastic B cells in the gastric tumor were alpha 4 beta 7-, CD62L+, whereas the intestinal secondary was alpha 4 beta 7+, CD62L-. Incubation of isolated tumor cells from the stomach by H. pylori generated T-cell-dependent proliferation of neoplastic B cells and induced expression of alpha 4 beta 7 integrin similar to the intestinal tumor. These observations indicate that reversal of homing receptor profile in the gastric tumor by antigen specific stimulation may be responsible for secondary intestinal dissemination. In normal stomach and normal MALT, alpha 4 beta 7 and CD62L expression reflected the differentiation of the B cell. Plasma cells were alpha 4 beta 7+, CD62L-, whereas a subset of memory B cells were alpha 4 beta 7-, CD62L+. Homing receptor expression in MALT lymphoma B cells was heterogeneous, however, in line with their memory B-cell phenotype in the majority of cases, the neoplastic B cells were alpha 4 beta 7-, CD62L+. Neoplastic plasma cells were always alpha 4 beta 7+, CD62L-. The venules in normal gastric mucosa expressed mucosal addressin cell adhesion molecule-1 but not peripheral lymph node addressin. In normal MALT, H. pylori-associated follicular gastritis and MALT lymphomas high endothelial venules coexpressed mucosal addressin cell adhesion molecule-1 and peripheral lymph node addressin. These findings suggest expression of lymphocyte homing receptors by B cells and vascular addressins by mucosal venules are similar in normal MALT and MALT lymphomas, and factors controlling normal mucosal B-cell traffic are also operational in MALT lymphomas. PMID- 9358763 TI - Cerebral lipid deposition in aged apolipoprotein-E-deficient mice. AB - To assess the influence of age and diet on cerebral pathology in mice lacking apolipoprotein E (apoE), four male apoE knockout mice (epsilon -/-), and five male wild-type (epsilon +/+) littermate controls were placed on a high-fat/high cholesterol diet for 7 weeks beginning at 17 months of age. All four aged knockout mice developed xanthomatous lesions in the brain consisting mostly of crystalline cholesterol clefts, lipid globules, and foam cells. Smaller xanthomas were confined mainly to the choroid plexus and ventral fornix in the roof of the third ventricle, occasionally extending subpially along the choroidal fissure and into the adjacent parenchyma. More advanced xanthomas disrupted adjoining neural tissue in the fornix, hippocampus, and dorsal diencephalon; in one case, over 60% of one telencephalic hemisphere, including nearly the entire neocortex, was obliterated by the lesion. No xanthomas were observed in aged wild-type controls fed the high-fat/high-cholesterol diet. Brains from 42 additional animals, fed only conventional chow, were examined; 3 of 15 aged (15- to 23-month-old) apoE knockout mice developed small choroidal xanthomas. In contrast, no lesions were observed in five young (2- to 4-month-old) apoE knockout mice or in any wild-type controls between the ages of 2 and 23 months. Our findings indicate that disorders of lipid metabolism can induce significant pathological changes in the central nervous system of aged apoE knockout mice, particularly those on a high fat/high-cholesterol diet. It may be fruitful to seek potential interactions between genetic factors and diet in modulating the risk of Alzheimer's disease and other neurodegenerative disorders in aged humans. PMID- 9358764 TI - Endogenous regulation of angiogenesis in the rat aorta model. Role of vascular endothelial growth factor. AB - The purpose of this study was to investigate the role of vascular endothelial growth factor (VEGF) in the rat aorta model of angiogenesis. Freshly cut aortic rings generated microvascular outgrowths in serum-free collagen gel culture. Angiogenesis was reduced to 10% when the explants were embedded in collagen 10 to 14 days after excision from the animal. Immunochemical studies of conditioned medium demonstrated secretion of VEGF by the aortic cultures. Levels of VEGF decreased during the second week of culture when the explants became quiescent and microvessels stopped growing. Treatment of quiescent aortic rings with exogenous VEGF stimulated angiogenesis and restored microvascular growth to values observed in cultures of freshly cut explants. Reverse transcriptase polymerase chain reaction of vasoformative collagen gel cultures of rat aorta demonstrated the expression of the alternatively spliced isoforms VEGF165, VEGF189, and the high affinity VEGF receptor flk-1. Reverse transcriptase polymerase chain reaction of rat aorta-derived cell strains confirmed the presence of VEGF165 and VEGF189 in endothelial cells, smooth muscle cells, and fibroblasts. The flk-1 receptor was expressed by endothelial cells but not by fibroblasts or smooth muscle cells, which is consistent with the endothelial target specificity of VEGF. The spontaneous angiogenic response of freshly cut aortic rings was inhibited by 70% with a neutralizing antibody against VEGF, whereas nonimmune IgG had no effect (P < 0.001). These findings provide evidence for a VEGF-mediated autocrine/paracrine regulation of angiogenesis in the rat aorta model. PMID- 9358765 TI - Transformation of rabbit vascular smooth muscle cells by human cytomegalovirus morphological transforming region I. AB - The association of human cytomegalovirus with atherosclerosis and the monoclonal hypothesis of atherogenesis suggested that transformation of vascular smooth muscle cells may be an outcome of the virus-host cell interaction. To test this hypothesis, rabbit aorta smooth muscle cells were transfected with the morphological transforming region I (mtrI) of human cytomegalovirus (HCMV) linked to the neomycin resistance gene. Foci of neomycin-resistant and morphologically transformed cells were isolated and expanded into fourteen RCMV strains. Eight of these strains acquired immortalization, but only one strain (RCMV-21) retained recombined viral sequences integrated in the cellular DNA. RCMV strains were heterogeneous in their morphology, expression of smooth muscle alpha-actin, growth, and mitogenic response to serum and fibroblast growth factor (FGF)-2 and 4. All RCMV strains assayed except RCMV-3 showed DNA synthesis in low serum medium and, with the exception of RCMV-1 cells, all showed a significant mitogenic response to FGF-2 and FGF-4, Maintenance of the transformed phenotype appeared independent of the retention of the transforming viral sequences, which was suggestive of a "hit-and-run" mechanism. These results suggested that morphological transformation by HCMV DNA sequences could enhance the mitogenic response of vascular smooth muscle cells to fibroblast growth factors. PMID- 9358766 TI - Platelets play an important role in TNF-induced microvascular endothelial cell pathology. AB - Tumor necrosis factor-alpha (TNF) is known to be an important mediator in the pathogenesis of several inflammatory diseases. Vascular endothelial cells represent a major target of TNF effects. Platelet sequestration has been found in brain microvessels during experimental cerebral malaria and lung in experimental pulmonary fibrosis, implying that it may participate in TNF-dependent microvascular pathology. In this study, we investigated the mechanisms of platelet-endothelial interaction, using co-cultures between platelets and TNF activated mouse brain microvascular endothelial cells (MVECs). Adhesion and fusion of platelets to MVECs was evidenced by electron microscopy, dye transfer, and flow cytometry. It was induced by TNF and interferon-gamma and depended on LFA-1 expressed on the platelet surface and ICAM-1 expressed on MVECs. The adhesion and fusion also led to the transfer of platelet markers on the MVEC surface, rendering these more adherent for leukocytes, and to an enhanced MVEC sensitivity to TNF-induced injury. These results suggest that platelets can participate in TNF-induced microvascular pathology. PMID- 9358767 TI - VEGF induces cardiovascular malformation and embryonic lethality. AB - The essential function of vascular endothelial growth factor (VEGF) in embryonic angiogenesis has clearly been documented in murine embryos with targeted deletions of either VEGF or its receptors. The effects of VEGF in the organogenetic phase of development have not been studied to date. Therefore, we applied 0.7 to 0.9 microgram of VEGF via methylcellulose carriers into the midbrain or onto the right forelimb of 4.5-day-old quail embryos. Another group of embryos was treated with 1 microgram of platelet-derived growth factor and controls were carried out using carriers without any growth factor. VEGF-induced cardiovascular malformations resulted in embryonic lethality. The venous area of the vasculature was dilated in almost all organs. The heart was most seriously affected and showed typical characteristics of insufficiency. VEGF strongly increased endocardial cell proliferation and obviously induced impairment of the growth rates of myocardium and endocardium. The myocardium of the ventricles was extremely thin, and septation defects were observed. As a result of the disturbed outflow, the atria were extremely dilated and thin-walled. The morphology of the hearts was reminiscent of that observed in congenital malformations such as Uhl's and Osler's syndromes. Our results show that expression of VEGF has to be tightly controlled during development. PMID- 9358768 TI - Non-small-cell lung carcinoma tumor growth without morphological evidence of neo angiogenesis. AB - Neoplastic growth is usually dependent on blood supply, and it is commonly accepted that this is provided by the formation of new vessels. However, tumors may be able to grow without neovascularization if they find a suitable vascular bed available. We have investigated the pattern of vascularization in a series of 500 primary stage I non-small-cell lung carcinomas. Immunostaining of endothelial cells has highlighted four distinct patterns of vascularization. Three patterns (which we called basal, papillary, and diffuse) have in common the destruction of normal lung and the production of newly formed vessels and stroma. The fourth pattern, which we called alveolar or putative nonangiogenic, was observed in 16% (80/500) of the cases and is characterized by lack of parenchymal destruction and absence of both tumor associated stroma and new vessels. The only vessels present were the ones in the alveolar septa, and their presence highlighted, through the whole tumor, the lung alveoli filled up by the neoplastic cells. This observation suggests that, if an appropriate vascular bed is available, a tumor can exploit it and grows without inducing neo-angiogenesis. This could have implications for strategies aimed at inhibiting tumor growth by vascular targeting or inhibition of angiogenesis. PMID- 9358769 TI - Modulation of the expression of integrins on glial cells during experimental autoimmune encephalomyelitis. A central role for TNF-alpha. AB - Integrins comprise a group of adhesion receptors involved in cell-cell and cell extracellular matrix interactions. Evidence is accumulating that integrins expressed on mononuclear cells play a central role in the induction of autoimmune diseases of the central nervous system. The effects of integrins on glial cell behavior, myelination, and angiogenesis suggest that they may also have a role in resolving inflammation in the nervous system and in promoting tissue repair. We investigated the temporospatial expression of integrins in the rat central nervous system during the course of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. A higher expression of alpha v- and beta 4 integrin subunits in astrocytes and alpha 2 integrin in oligodendrocytes was observed in active lesions of experimental autoimmune encephalomyelitis, in comparison with controls. Proinflammatory cytokines, primarily TNF-alpha, also enhanced alpha v, beta 4, and alpha 2 expression in purified glial cells ex vivo. Furthermore, we observed that the expression of some integrin subunits was modulated in the cerebral vasculature during inflammation. Our results suggest an active role for glial and vascular integrins in the regulation of autoimmune diseases of the central nervous system, opening an avenue for new potential immunotherapies. PMID- 9358771 TI - Genetic progression and heterogeneity in intraductal papillary-mucinous neoplasms of the pancreas. AB - Intraductal papillary-mucinous neoplasms (IPMNs) of the pancreas are ideal neoplasms to study clonal progression and genetic diversity because of their large size and prominent intraductal component. We microdissected 55 histologically defined areas from 13 IPMNs, extracted the DNA from each, and performed polymerase chain reaction (PCR)-based microsatellite analysis to detect loss of heterozygosity on chromosome arms 1p, 3p, 6q, 8p, 9p, 17p, 18q, and 22q. LOH was identified at 1p in two cases, at 3p in four cases, at 6q in seven cases, at 8p in four cases, at 9p in eight cases, at 17p in five cases, at 18q in five cases, and at 22q in one of the IPMNs examined. In one of the IPMNs, the allelic losses were uniform throughout multiple microdissected areas, and in four of the IPMNs, there was evidence of clonal progression. In contrast, in three of the IPMNs, substantial allelic heterogeneity was seen. This remarkable heterogeneity may, in part, be due to the slow growth rate of these neoplasms. PMID- 9358770 TI - Mononuclear phagocyte hydrolytic enzyme activity associated with cerebral HIV-1 infection. AB - In patients with HIV encephalitis, activated macrophages and microglial cells in the brain are infected by the human immunodeficiency virus (HIV-1). Immune activation can release neurotoxic chemicals including cytokines, free radicals, autocoids, and hydrolytic enzymes. In this study, the presence of hydrolytic enzymes in acquired immune deficiency syndrome (AIDS)-related neurodegeneration was addressed. Activities of four lysosomal hydrolases were assayed in the frontal lobe of 69 males who died with AIDS and 31 age-matched control men. Activities of all four enzymes were increased significantly (1.6 to 3.6 times) in white matter of patients with AIDS. Less pronounced increases were present in cerebral cortex. Of 69 of the subjects with AIDS, 50 (72%), had at least one abnormally active enzyme. Patients with HIV encephalitis and other neuropathological changes were affected as were many subjects without any clear neuropathological anomaly. Lysosomal cathepsin D immunostaining revealed increased lysosomes within perivascular macrophages, multinucleated cells, activated microglial cells, and hypertrophic astrocytes. Increased enzyme activity was correlated significantly with assay results for HIV-1 DNA using the polymerase chain reaction. The release of acid hydrolases associated with cerebral HIV-1 infection could lead to unopposed hydrolysis of matrix and surface proteins. These post-translational disturbances could contribute to white matter and synaptic injury in AIDS. PMID- 9358772 TI - Expression of the complement regulatory proteins decay accelerating factor (DAF, CD55), membrane cofactor protein (MCP, CD46) and CD59 in the normal human uterine cervix and in premalignant and malignant cervical disease. AB - The membrane-bound complement regulators decay-accelerating factor (DAF, CD55), membrane cofactor protein (MCP, CD46), and CD59 are broadly expressed proteins that act together to protect host tissues from autologous complement. Comparison of expression profiles of these proteins between normal and pathological tissues could reveal a mechanism by which tumor cells evade complement-mediated killing. Expression of the regulators was therefore examined in the normal human uterine cervix, in cervical intraepithelial neoplasia (CIN; n = 23), and in cervical squamous carcinomas (n = 6). DAF and MCP were reciprocally expressed in normal ectocervical epithelium. MCP was confined predominantly to the basal and parabasal layers with more extensive expression in metaplastic squamous epithelium. An apparent expansion in MCP expression was observed in more severe premalignant lesions whereas cervical carcinoma were uniformly MCP positive. By contrast, DAF expression appeared unaltered in premalignant lesions and variable in carcinomas. However, increased DAF was observed in stromal cells directly adjacent to infiltrating tumor cells. A low molecular weight DAF product was detected in tumors, and preliminary evidence suggests this may be derived from stromal cells. Overall, changes in expression of C3 convertase regulators in both the stromal and epithelial compartments may be important for evasion of immune surveillance in cervical cancer. PMID- 9358773 TI - Keratinocyte growth factor in inflammatory bowel disease. Increased mRNA transcripts in ulcerative colitis compared with Crohn's disease in biopsies and isolated mucosal myofibroblasts. AB - Inflammation in the gastrointestinal tract is associated with increased epithelial cell proliferation. Keratinocyte growth factor (KGF) is an epithelial cell mitogen widely expressed by mesenchymal cells subjacent to the epithelial cells. In this study, we have investigated the expression and distribution of KGF in normal and diseased (Crohn's disease and ulcerative colitis(UC)) intestine by quantitative competitive reverse-transcriptase polymerase chain reaction in whole biopsies and purified lamina propria myofibroblasts and by in situ hybridization. Analysis of whole mucosal biopsies reveals significantly higher numbers of KGF mRNA transcripts in UC compared with Crohn's colitis and control colon (P < 0.001). KGF transcripts were also elevated in Crohn's ileitis compared with normal ileum. In situ hybridization showed a marked increase in cells expressing KGF mRNA throughout the lamina propria in both UC and Crohn's tissue. In Crohn's disease, positively hybridizing cells were only rarely seen in the submucosa but were abundant around the bases of the crypts and were not associated with lymphoid aggregates. In purified mucosal myofibroblasts, increased (15- to 20 fold) KGF mRNA expression was seen in UC compared with control and Crohn's tissue. These results confirm and extend earlier studies showing that KGF transcripts are elevated in inflammatory bowel disease, but they show for the first time that transcripts are higher in UC than Crohn's disease because of increased production by mucosal mesenchymal cells. PMID- 9358775 TI - Routine ultrasonography in the diagnosis of acute appendicitis: a valuable tool in daily practice? AB - Initial clinical examination, laboratory inflammation parameters, and routine ultrasonography (US) were evaluated prospectively in 205 patients admitted for suspected acute appendicitis. The purpose of the study was to compare initial clinical examination and laboratory tests with the accuracy of US obtained in daily practice. All ultrasonographic examinations were performed by residents in radiology and radiologists qualified for routine abdominal US but not specifically trained in the diagnostics of appendicitis. Initial clinical examination had the highest sensitivity, but the specificity was poor. The single laboratory tests were of limited value in predicting appendicitis. The accuracy of US was disappointing, and the reported promising results of previous studies were not confirmed. Sonographers with less experience had a higher sensitivity but a poorer specificity compared with more experienced sonographers, who had a high specificity at the cost of extremely poor sensitivity. Biases, dichotomization problems, and factors influencing the accuracy of US in patients with suspected acute appendicitis are discussed. PMID- 9358776 TI - Carcinoma of the common bile duct with superficial spread to the intrahepatic segmental bile ducts: a case report. AB - A 54-year-old woman presented with jaundice. Percutaneous transhepatic biliary drainage, cholangiography via a percutaneous transhepatic biliary, drainage catheter, and percutaneous transhepatic cholangioscopy were performed to alleviate the jaundice and to evaluate her biliary system. A diffuse-type tumor was detected in the common bile duct. The tumor had spread superficially up to the right anterior segmental duct and the left hepatic duct and involved the caudate branches. Curative surgery, which included a right anterior segmentectomy, total caudate lobectomy, and pylorus-preserving pancreatoduodenectomy, was performed. The histopathologic diagnosis was moderately differentiated tubular adenocarcinoma originating at the common bile duct. The extent of the superficial spread of the tumor corresponded to our preoperative determination. Her postoperative recovery was uneventful. In this case report, we discuss the accurate preoperative diagnosis and rational surgical treatment of bile duct carcinoma with superficial spread. PMID- 9358774 TI - Subependymal astrocytic hamartomas in the Eker rat model of tuberous sclerosis. AB - Tuberous sclerosis (TSC) is an autosomal dominant syndrome that is linked to two genetic loci: TSC1 (9q34) and TSC2 (16p13). Brain manifestations such as cortical tubers and subependymal hamartoma/giant cell astrocytomas are major causes of TSC related morbidity. In this study, we describe the central nervous system involvement in a unique rodent model of tuberous sclerosis. The Eker rat carries a spontaneous germline mutation of the TSC2 gene and is predisposed to multiple neoplasia. In a series of 45 adult Eker carriers (TSC2 +/-), three types of focal intracranial lesions were found, of which the subependymal and subcortical hamartomas were most prevalent (65%). There exist remarkable phenotypic similarities between the Eker rat and human subependymal lesions. Our study indicates that the predominant cellular phenotype of the subependymal hamartomas is astroglial and suggests that the neuronal contribution within these lesions is, in part, the result of pre-existing myelinated axons. The hamartomas did not show evidence of loss of the wild-type TSC2 allele; it remains to be determined whether TSC2 inactivation is necessary for their pathogenesis. This genetically defined rodent model may be useful in elucidating the molecular and developmental basis of the subependymal giant cell astrocytoma in humans. PMID- 9358777 TI - Yolk sac tumor of the anterior mediastinum: the role of palliative surgery. AB - Primary endodermal sinus tumor (yolk sac tumor) of the mediastinum is a very rare extragonadal germ-cell neoplasm. Most patients are young and male. Since initial description of the endodermal sinus tumor in 1959, this neoplasm has been found more and more frequently. alpha-Fetoprotein is an important tumor marker. The treatment consists of multimodal therapy. A combined approach with chemotherapy followed by surgical resection of residual tumor seems to be the optimal management of this tumor. Even a palliative surgical resection in advanced tumors is indicated. We describe the case of a patient with an advanced yolk sac tumor that was refractory to chemotherapy. This patient was treated by palliative surgical resection. PMID- 9358778 TI - Primary hepatic lymphoma: unusual presentation and clinical course. AB - Although primary hepatic lymphoma is rare, it should be considered in the differential diagnosis of a hepatic tumor, because it is usually associated with a favorable prognosis. This report describes an unusual case of primary hepatic lymphoma with an atypical presentation (only mild, right upper quadrant pain and no hepatomegaly) followed by acute fulminating hepatic failure, metabolic acidosis, followed by a rapidly fatal course. A review of the literature and discussion of the disease are also presented. PMID- 9358779 TI - Parathyroid carcinoma: diagnosis and management. AB - A patient with severe hypercalcemia and a palpable neck mass is presented. The highest calcium was 18.8 mg/dL. A left lower neck mass was felt on examination. The trachea was deviated to the right side on a chest film. A barium swallow demonstrated an indentation on the left side of the esophagus. An en-bloc resection of the mass including the thyroid lobe, the strap muscles, and the recurrent laryngeal nerve was done. The pathologic specimen revealed parathyroid carcinoma with dense fibrous septae, invasion of the capsule, and vascular invasion. The patient is alive and without evidence of hypercalcemia or recurrence of the disease 23 years after surgery, probably the longest survivor with carcinoma of the parathyroid gland. Parathyroid carcinoma should be suspected in any patient with severe hypercalcemia and a palpable mass. The best chance for cure is obtained by performing a wide surgical excision during the initial operation. PMID- 9358780 TI - Cystic pancreatic neoplasms: report of a case and review of the literature. AB - Cystic neoplasms of the pancreas are rare tumors. Because of very limited experience even in large medical centers, there is much debate regarding the evaluation and management of patients with these tumors. Recently, a patient presented to our community teaching hospital with an unusual complication of this rare tumor. She was found to have intra-abdominal hemorrhage from the erosion of a serous cystadenoma into the surrounding vasculature. In this report, we present this case and review the current literature with regard to the presentation, diagnosis, and management of cystic neoplasms of the pancreas. PMID- 9358781 TI - Massive hemorrhage from proximal gastric vascular ectasias. AB - Gastric vascular ectasias are rare lesions of the gastric antrum that may be the cause of iron deficiency anemia in the elderly. We report a case of proximal gastric vascular ectasias in a young man which caused rapidly fatal hemorrhage. This case is instructive because of the patient's youth, the location of the lesions, and the rapidly fatal hemorrhage. PMID- 9358782 TI - Pyloric exclusion in the management of duodenal trauma: is concomitant gastrojejunostomy necessary? AB - Pyloric exclusion with gastrojejunostomy (PE-GJ) has been recommended in patients with severe injuries to the pancreatoduodenal complex. Recently, the management philosophy for pancreatoduodenal injuries has been that less treatment is probably the best treatment. But whether gastrojejunostomy (GJ) should be used routinely with pyloric exclusion (PE) remains controversial. A retrospective review was conducted of patients who underwent PE at a Level I trauma center during a 36-month period. Forty-five patients had duodenal injuries and 12 of these (27%) underwent PE for management of complex duodenal injuries. Gunshot wounds were the cause of the injuries in 10 of the 12 patients (83%). Eight patients (67%) underwent PE-GJ and had a mean hospital stay of 25 days. Four patients (33%) underwent PE alone and had a mean hospital stay of 29 days. All 12 patients had spontaneous opening of the PE, regardless of the technique used. One patient (12.5%) in the PE-GJ group developed marginal ulceration and significant hemorrhage, and one patient died in the PE-GJ group. The reported incidence of marginal ulceration in the PE-GJ group, the spontaneous opening of the pylorus, and the need to limit the extent of surgical repair to focus on all other associated lesions present in these patients, suggest that GJ should not be used routinely in patients undergoing PE for the management of severe pancreatoduodenal injuries. PMID- 9358783 TI - The etiology of the adult indirect inguinal hernia: revisited. AB - It has generally been historically stated that indirect inguinal hernias develop only in patients who have a patient processus vaginalis that enlarges to become a hernia sac. Occasionally, this theory has been challenged but without any objective evidence. Herniography was performed by placing 50 mL of nonionic contrast material into the peritoneal cavity. The patient was then placed in a prone position with the head of the table elevated. Films of the inguinal fossae were obtained with the patient straining. The herniogram revealed a right indirect inguinal hernia. There was no left inguinal hernia, nor was there a patent processus vaginalis on the left side. Two years later, the patient developed left inguinal discomfort and swelling and was found to have a moderate sized left inguinal hernia. At the time of operation, an indirect sac of moderate size was present. A mesh plug repair was performed. This case report is the first published objective evidence that, contrary to common thought, a patent processus vaginalis is not a necessary prerequisite to the development of an indirect inguinal hernia. PMID- 9358784 TI - Internal iliac revascularization during aortic aneurysm replacement: a review and description of a useful technique. AB - The coexistence of infrarenal aortic aneurysm and internal iliac artery aneurysm may represent a management problem with regard to preservation of the pelvic blood supply. In this article, we review the methods available for maintaining the pelvic blood flow and describe a useful technique that we have successfully utilized in seven patients to preserve the hypogastric artery blood flow. PMID- 9358785 TI - Torsion of the gallbladder: a case report and review of the literature. AB - Acute torsion of the gallbladder is a rare and poorly understood entity. Since it was first described by A.V. Wendel in 1898, approximately 300 cases have been reported in the literature. The treatment of choice remains immediate cholecystectomy, and most cases are diagnosed intraoperatively. There are typical clinical and radiological findings consistent with torsion of the gallbladder that should raise the index of suspicion for this condition preoperatively. We present a case of gallbladder torsion and discuss the pertinent literature. A re emphasis is placed on the salient clinical features, and the availability of diagnostic tests is stressed. Given the possibility of laparoscopic cholecystectomy and the increasing incidence with which gallbladder torsion is being witnessed today, the importance of a preoperative suspicion is discussed. PMID- 9358786 TI - Supraglottic and subglottic airway injury due to deployment and rupture of an automobile airbag. AB - A 46-year-old white female with underlying interstitial lung disease and asthma was driving a pickup truck when it was struck broadside. The airbag inflated and then ruptured, forcing inhalation of its contents. This produced facial desquamation, productive cough, wheezing, and headache. Chest radiograph showed bilateral interstitial fibrosis. Pulmonary function tests demonstrated small airway obstruction, hyperinflation, and impaired diffusion. Computerized tomography showed extensive sinusitis. She improved following treatment with inhaled steroids, bronchodilators, and oral antibiotics. Inhalation of the airbag contents produced supraglottic and subglottic airway inflammation, resulting in sinusitis and exacerbation of her underlying asthma. PMID- 9358787 TI - Choledochal cyst and neoplasm: a comprehensive review of 106 cases and presentation of two original cases. AB - Choledochal cysts are prone to complications: cholangitis, biliary cirrhosis, portal hypertension, lithiasis, rupture, pancreatitis, and carcinoma. The coincidence of choledochal cysts and neoplasia ranges from 2.5 to 26 per cent. One hundred six cases of choledochal cysts with neoplasms have been collected from the literature. We have tabulated the results of 68 cases found to have a neoplasm at the initial laparotomy and of 38 patients with pristine choledochal cysts treated electively who subsequently developed a neoplasm. Two original cases are presented. The primary site of neoplasia was not confined only to choledochal cysts. There appears to be a propensity for malignancy to develop anywhere in the biliary tract or gallbladder or pancreas in conjunction with the choledochal cyst. Accompanying choledochal cysts is a high incidence of an anomalous relation at the pancreaticobiliary junction with subsequent malignancy formation. A pathogenetic basis is postulated. PMID- 9358788 TI - Stereotactic breast biopsy: comparison of 14- and 11-gauge Mammotome probe performance and complication rates. AB - The purpose of this study was to compare specimen weights, tissue acquisition times, and complication rates for 14- and 11-gauge Mammotome probes during stereotactic breast biopsy. Three hundred forty stereotactic Mammotome breast biopsies were reviewed for complications; 269 (79%) of these biopsies were performed with the 14-gauge Mammotome probe, and 71 (21%) with the 11-gauge probe. Aggregate specimen weights and tissue acquisition times were measured in 248 out of 340 stereotactic Mammotome breast biopsies. Of these 248 lesions, 186 (75%) were performed with the 14-gauge probe, and 62 (25%) with the 11-gauge probe. Complication rates were essentially the same for each probe: 1.1 per cent for the 14-gauge probe and 1.4 per cent for the 11-gauge probe (not significant). More total aggregate breast tissue was obtained during the biopsies performed with the 11-gauge Mammotome probes than with the 14-gauge probes (1730 versus 1067 mg; P < or = 0.0001). The number of specimens acquired was smaller during the 11-gauge biopsy sessions (18 versus 27; P < or = 0.0001). The average per specimen weight was significantly higher with the 11-gauge probe: 96 versus 40 mg per specimen (P < or = 0.0001). The average tissue harvesting time was approximately the same with each probe size, 15.6 minutes for the 11-gauge probe and 15.4 for the 14-gauge probe (not significant). Consequently, the harvesting rate (mg of tissue obtained per minute) was 111 mg/minute with the 11-gauge probe versus 69 with the 14-gauge probe (P < or = 0.0001). The per-specimen breast tissue yield, using the 11-gauge probe, was more than two times greater than the yield using the 14-gauge probe. There was no increase in procedure time, and there were no additional complications. PMID- 9358789 TI - Laparoscopy-assisted small bowel resection. AB - The increased use of laparoscopy in the management of gastrointestinal problems continues to expand. Procedures such as jejunostomies, diagnosis of intestinal obstruction or ischemia, resection of the small bowel, and lysis of adhesions can be managed with this technique. We present our experience with four cases undergoing laparoscopic resection of the small bowel. The mean age of the three males and one female was 55 years. The operative procedure was performed under general anesthesia with complete laparoscopic exploration of the abdominal cavity. The type of pathology and extent of disease was defined: one had leiomyoma, two had unspecific ileitis, and one had metastatic breast cancer. This was followed by exteriorization and resection. Patients were allowed to have a liquid diet the day of surgery. The average hospital stay was 3 to 4 days. The mean intraoperative time was 124 minutes. No postoperative complications were observed. Laparoscopic small bowel resection can be performed expeditiously and with minimal morbidity, allowing accurate diagnosis and treatment of these conditions. PMID- 9358790 TI - The utility of ultrasonography in the evaluation of groin masses: a case report. AB - Surgeons are often faced with the evaluation and management of groin masses. In most cases, an accurate history and physical examination will establish a diagnosis. Ultrasonography is being increasingly used in the evaluation of surgical problems. A case is presented in which bedside ultrasound was utilized in the evaluation of a large, symptomatic left inguinal mass found to be a synovial cyst on exploration. The differential diagnosis of cystic groin lesions and the impact of ultrasonography on diagnosis is reviewed with emphasis on synovial cysts. Surgeon-directed ultrasonography is an asset in the diagnosis of some patients with groin masses and may assist in the identification of those lesions requiring prompt operative intervention. PMID- 9358791 TI - Optimal diagnosis of splenic vein thrombosis: brief clinical report. AB - The presence of splenic vein thrombosis is sometimes very difficult to diagnose. We present a patient in whom the splenic vein was thought to be patent by ultrasound and conventional celiac angiography. Because of high clinical suspicion and continued bleeding, he underwent a selective intra-arterial digital splenic angiogram. The venous phase clearly showed proximal (hilar) splenic vein occlusion with filling via collaterals in real time. Splenectomy confirmed the diagnosis. We believe that a selective intra-arterial digital splenic angiogram is the radiographic study of choice for suspected splenic vein thrombosis. PMID- 9358792 TI - Gastrosplenic fistulas: a case report and review of the literature. AB - A case is presented of a patient who developed a gastrosplenic fistula during a course of chemotherapy for differentiated histiocytic lymphoma. The fistula was observed during upper gastrointestinal endoscopy (gastroscopy) and confirmed by CT scan with contrast. The fistula was followed endoscopically and noted to have closed spontaneously with confirmed closure at laparotomy. The clinical management of this complication is discussed, and the literature pertaining to this rare condition is reviewed. PMID- 9358793 TI - Malignant pleural mesothelioma diagnosed after chest trauma. AB - Two patients who presented with persistent pulmonary symptoms after chest trauma and were diagnosed to have malignant pleural mesothelioma are described. The symptoms, more than expected from trauma, prompted earlier diagnosis of this underlying disease. The possibility of unknown preexisting diseases should always be considered in posttraumatic patients with unusual presentations. PMID- 9358794 TI - Heparin versus citrate regional anticoagulation during autotransfusion in a porcine intra-abdominal hemorrhage model. AB - Our objective was to determine the effects of anticoagulants and blood loss on hemodynamic, hematologic, and coagulation parameters following autotransfusion in an animal model of intraabdominal hemorrhage. We performed a prospective, randomized observational animal study at an animal research laboratory at a university medical center. Eight Landrace, domestic pigs, weighing 17-23 kg, each underwent jugular venous and iliac arterial catheterization and laparotomy with retroperitoneal dissection for aortic exposure to simulate an operative environment. Following baseline laboratory and hemodynamic determinations, intra abdominal hemorrhage was accomplished via aortotomy in three sequential 10 mL/kg blood volumes. After allowing pooling in the exposed retroperitoneum to ensure tissue contact, the shed blood was suctioned, processed, and washed in an autotransfusion device utilizing either heparin (n = 4) or acid-citrate-dextrose (n = 4) as a system anticoagulant. Prior to autologous transfusion, each pig received a 20 mL/kg intravenous bolus of 0.9 per cent saline to treat shock. The processed blood was then infused, and laboratory and hemodynamic measurements were repeated following each cycle of hemorrhage and autotransfusion. Sequential fixed volume hemorrhage resulted in significant reductions in mean arterial pressure. Despite crystalloid infusion and transfusion of processed shed blood, postresuscitation mean arterial pressure did not return to baseline values, with no difference noted between anticoagulant groups. Infusion of increasing volumes of autologous blood resulted in significant reductions in hematocrit, platelet count, fibrinogen, antithrombin III, ionized calcium, and total protein. The decrease in concentration of each variable was independent of the choice of anticoagulant with the exception of antithrombin III, with higher levels noted in animals receiving blood anticoagulated with acid-citrate-dextrose. Prothrombin time and partial thromboplastin time were unaffected by volume of autologous transfusion or choice of anticoagulant. We conclude that changes in hemodynamic, hematologic, and coagulation parameters associated with hemorrhage and autotransfusion appear related more to the volume of blood loss and the cumulative pheresis of plasma than to the choice of anticoagulant. PMID- 9358795 TI - Fournier's gangrene: review of fifteen cases. AB - Fournier's gangrene is a synergistic necrotizing fasciitis of the perineum and abdominal wall along with the scrotum and penis in men and the vulva in women. The process was believed to be idiopathic in initial descriptions. Fifteen patients were treated for Fournier's gangrene between 1990 and 1995 in the Departments of General Surgery and Urology, School of Medicine, Ataturk University, Erzurum, Turkey. The most common causes were perianal sepsis and urogenital diseases. Escherichia coli and Staphylococcus aureus were identified most commonly in cultures of necrotic tissue. The mortality rate was 20 per cent despite aggressive surgical debridement and broad-spectrum antibiotics. PMID- 9358797 TI - A simple method of feeding jejunostomy tube placement. PMID- 9358796 TI - Balloon tamponade: an alternative in the treatment of liver trauma. AB - Penetrating hepatic injury remains a therapeutic challenge for the surgeon. Surgical technique for the management of penetrating hepatic trauma includes balloon tamponade, which is most useful for central gunshot wounds that pass through both lobes. Our patient had two entrance wounds and no exit wounds. However, a bullet was palpable in the subcutaneous tissue just beneath the right shoulder in the supraclavicular region. A Penrose drain and red rubber catheter balloon device were placed in the abdominal cavity. The balloon device was inflated for 48 hours, after which the Penrose drain was allowed to slowly deflate and was removed. The abdomen was re-explored, found to be negative, and then closed. The patient's postoperative course was uneventful, and he was discharged on postoperative day 14. The use of balloon tamponade is an option that should be kept in the surgeon's armamentarium for use in selected patients with hepatic trauma. PMID- 9358798 TI - Observations on early suture materials: the first stitch in time. PMID- 9358799 TI - Classification and grading of chronic venous disease in the lower limbs. A consensus statement. Ad Hoc Committee, American Venous Forum. AB - Advances in modern technology have made available a large number of both invasive and non-invasive investigations that can provide information not only on the presence, absence or anatomic extent but also quantitation of the abnormalities. It was soon realised that the combinations of presence, absence, extent and severity of reflux and/or obstruction in the deep, superficial or perforating veins are so large that the classification of chronic venous disease would be a major challenge. This challenge has been taken up by the Consensus Committee which met in Maui on 22-26 February 1995. The Consensus that developed is in three parts. Part I deals with a classification system that covers the Clinical picture, the Etiology, the Anatomic distribution and the Pathophysiology (the CEAP Classification). Part II suggests a scoring system intended to be evaluated on patients classified according to the CEAP classification, and Part III provides guidelines on the use of various investigations which according to the current medical literature will aid in the classification. The Consensus Committee has retained the copyright of the document so that it can be made available for reproduction to all interested parties. Reproduction is free provided there is no alteration and it is always reproduced in its entirety. The development of this consensus document is a continuous process. Suggestions for improvement are expected from all involved in the study and management of patients with chronic venous disease. The classification has already been presented and debated at several national and international meetings and plans for an updated version are already in progress. PMID- 9358800 TI - Ultrastructural demonstration of mast cells in varicose veins of lower limbs: presence of mast cell-mediated mechanism. AB - OBJECTIVE: Mast cells are suggested to play an essential role during development of varices in the lower limbs. EXPERIMENTAL DESIGN: Investigation of the ultrastructure of mast cells in varicose lesions. SETTING: Saga Medical School, Yamamoto Surgical Hospital. PATIENTS: Eighteen varicose veins of 17 patients and 9 normal saphenous veins of 9 patients were examined. Patients who had undergone sclerotherapy for varices were excluded. INTERVENTIONS: Radical stripping surgery was performed on all varicose veins. MEASURES: Ultrastructural observations. RESULTS: In normal saphenous veins, mast cells usually singly embedded in dense collagen bundles as resident cells. They have characteristic crystalline granules of storage type. In varicose veins, mast cells show different features such as an increase of altered granules of discharging type, degranulation and intimate relationship with fibroblasts and lymphocytes. CONCLUSIONS: The observations suggest the presence of mast cell-mediated mechanism by releasing some mediators in the development of varices. PMID- 9358801 TI - Angiomegaly. AB - The authors report a clinical and ultrastructural study on a group of patients with angiomegaly, a vascular disorder characterized by elongated and distended blood vessels affecting the arterial (arteriomegaly) and/or venous system (venomegaly). The arterial group, drawn from a large arteriographic series, focuses on a comparison between atherosclerotic arteriopathy and arteriomegaly. The venous group, drawn from a large ultrasound series of vein disorders, is made up of patients with venomegaly. Venomegaly gives rise to few or no symptoms and it appears to be less frequent than arteriomegaly but as the latter proved to be associated in the majority of cases studied. Based on ultrastructural findings, the chief abnormality of angiomegaly seems to lie in a specific alteration of the elastic component of the vessel wall. We found slightly osmiophil amorphous elastic material neighbouring the basement membrane of the myocytes of the vessel walls. In the superficial parts of these myocytes occurred a great number of pinocytotic vesicles indicating for a rich creation of the new elastic material. Middle or highly osmiophil thick elastic fibers with irregular side protrusions were also found among myocytes remembering the moth-eaten picture. Results from a large ultrasonographic study on patients' relatives suggest an inheritability of this vascular disorder. PMID- 9358802 TI - Secondary aortoduodenal fistulae. AB - Secondary aortoenteric fistula (SAF) is a rare but fatal complication of reconstructive aortoiliac surgery. The prevention, diagnosis and treatment of this complication remains a challenging problem in everyday practice. Nine cases of secondary aortoduodenal fistulae during the period of 1980 to 1992 are presented. Their main symptom was bleeding of the upper gastrointestinal tract. The mean time interval since the aortic surgical procedure was 32 months. Removal of the old graft and closure of the duodenal defect was the first stage of the operative procedure. One patient underwent replacement of the old graft, with a new graft, while in the remaining three patients extranatomical bypass was not necessary because of satisfactory circulation in the lower extremities. In five patients extranatomical revascularization of the lower limbs was performed postoperatively at various intervals. Three patients died postoperatively. Follow up of the remaining patients ranged from one month to 8 years. Bleeding of the upper gastrointestinal tract in patients with a history of intrabdominal reconstructive vascular surgery must raise severe suspicion as to the certainty of existance of SAF unless the diagnostic procedure, mainly exploratory laparotomy, excludes this possibility. PMID- 9358803 TI - Endogenic non-enzymatic antioxidative system of polyester grafts during their healing. AB - OBJECTIVE: Non-enzymatic low-molecular antioxidants are one of the important mechanisms which protect cells against the toxic effect of oxygen. The aim of the present study was to determine the content of glutathione, glutathione reductase, and ascorbic acid in the principal layers of polyester grafts. INTERVENTIONS AND MEASURES: The experiments were carried out on 24 mongrel dogs, in which polyester double velour DALLON grafts were implanted. Seven days, 1, 4, and 12 months after the operation the grafts were excised. The following were determined: glutathione content by use of a GSH-400 system, glutathione reductase activity by the method of Langdon and Mize, and ascorbic acid content by the Kyaw method. RESULTS: It was found that the glutathione content in the graft neointima was 33% lower, in the graft neomedia higher during the first 4 months, in the neoadventitia 50% lower after 4 months than in the corresponding layers of the aorta (p < 0.01). The activity of glutathione reductase was significantly higher in all the graft layers during 12 months' observation than in the normal aorta layers. The ascorbic acid content of the graft layers was lowest 7 days after the implantation, and then in time increased so that 12 months after the operation it reaches its highest values. CONCLUSIONS: Our study shows that low non-enzymatic antioxidative potential is not capable of proteoting the newly forming graft layers, particularly the neointima, against oxygen toxicity. Thus, it would be beneficial to administer antioxidants (vitamin C, vitamin E, and N acetylcysteine). PMID- 9358804 TI - Metabolic alterations of skeletal muscle during ischaemia and reperfusion. AB - Recent studies have demonstrated that skeletal muscle cells are resistant to prolonged periods of ischaemia, but damage is observed after reperfusion. Periods of time longer than three hours of normothermal ischaemia in skeletal muscle lead to irreversible lesions. In the present study muscle metabolism during ischaemia and reperfusion was studied. After three hours of ischaemia two experimental groups were produced depending on whether or not they were to be followed by two hours of reperfusion. Adult mongrel dogs were submitted to ischaemia of the gracilis muscle. In this tissue, energetic metabolism was evaluated by its mitochondrial function and by glycogen level measurement. In a second experimental group the same ischaemic period was followed by two hours of reperfusion. The contralateral muscle of the same animal was used as a control. No changes in mitochondrial function, analysed by respiratory control ratio (RCR) or in any of its components, basal (state IV respiration) or ADP-activated (state III respiration) was observed. Glycogen levels also remained unaffected during the three hour ischaemic period and after two hours of reperfusion. We conclude that in the present dog model of gracilis preparation the skeletal muscle displays great resistance to ischaemia and reperfusion. PMID- 9358805 TI - The diagnosis and management of splanchnic artery aneurysms. Report of 8 cases. AB - The purpose of this report is to describe the clinical characteristics and surgical technique for splanchnic artery aneurysms. Over the past 10 years we have surgically resected 8 cases of splanchnic artery aneurysms including 2 cases involving the superior mesenteric artery, 3 involving the renal artery, 1 involving the hepatic artery and 2 involving the splenic artery. Diagnosis was established preoperatively in all patients by splanchnic angiography. Surgical treatment for splanchnic artery aneurysms is indicated in any symptomatic patient, in all symptomatic patients with suspected renal aneurysmal expansion and in patients who have renal aneurysms occurring with functionally important renal stenosis, usually associated with hypertension, in all patients with surgical accessible hepatic artery aneurysms, in all patients who have superior mesenteric artery aneurysms having a high tendency to rupture, and in all patients with an asymptomatic splenic artery aneurysms 1.5 cm or larger in diameter. Although splanchnic artery aneurysms are uncommon and asymptomatic, we recommend that splanchnic arteries should be treated surgically because of their tendency to rupture or organ failure. PMID- 9358806 TI - Modified "elephant trunk" procedure obliterating the false lumen in aortic dissection. AB - We describe a new application of the "elephant trunk" procedure for repairing aortic dissection. The false lumen of the distal aorta is obliterated between an external sheath of Teflon felt and an internal Dacron graft used as the "elephant trunk". This simple technique makes possible secure anastomosis with the distal friable tissue, and prevents suture hole leaks. PMID- 9358808 TI - Defibrillation threshold and electrode configurations: an experimental study testing three configurations in twelve pigs. AB - OBJECTIVE: The choice between epicardial or subcutaneous patches remains unclear and depends essentially on the team's habits. This study tested how much an additional patch can decrease defibrillation threshold (DFT), and compared a Subcutaneous Array and an epicardial patch. Today most implantable automatic defibrillators have a transvenous endocardial configuration alone but when the DFT remains high an additional patch is necessary. EXPERIMENTAL DESIGN: Three different configurations were tested with biphasic shocks in 12 pigs: Endovenous lead (Endo), Endovenous lead + subcutaneous patch (Endo + SQ) and Endovenous lead + epicardial patch (Endo + Epi). For each animal DFTs were determined in a balanced random order for the 3 configurations. Ventricular fibrillation was induced by pacing (30 Hz, 8 V, for 5 seconds). DFT was accurately measured with the up/down procedure until 3 reversal of defibrillation success or failure occurred. DFTs (mean +/- SD) were extracted and compared using paired t test and analysis of variance. RESULTS: DFTs were 14.6 +/- 11.0 J for Endo and significantly decreased (p < 0.01) when an additional SQ (9.4 +/- 7.2 J) or epicardial patch (8.9 +/- 6.5 J) was added to endovenous lead, without significant difference between Endo + SC and Endo + Epi configurations. CONCLUSIONS: Regarding this important decrease of DFT (respectively -35% for Endo + SC and -39% for Endo + Epi), additional patches should be more often recommended in patients with low safety margin of DFT. In those cases subcutaneous patches should be preferred instead of epicardial patches. Moreover, an additional reason to recommend this attitude could be the possible generator batteries saving. PMID- 9358807 TI - Left-sided inferior vena cava. Report of a case occasionally encountered while performing an aorto bifemoral bypass and review of the literature. AB - We present a case of left-sided inferior vena cava unexpectedly observed during an operation of aorto bifemoral bypass in a patient with severe Leriche syndrome and almost complete obstruction of the infrarenal aorta. This very rare congenital malformation (0.2-0.5%) was not recognized by the duplex scanner performed preoperatively, probably because of the low level of suspicion carried on by an experienced operator. AngioCT or angioMR, which would have surely shown us the anomaly, were not done because, in the lack of an aneurysmal disease or other abdominal pathological situations, these investigations were not required before operation. The possible hazards of such an unrecognized malformation are great, mostly in terms of incontrollable intraoperative hemorrhages, but the final outcome of this case was positive. PMID- 9358809 TI - Increased risk of coronary artery bypass grafting for left ventricular dysfunction with dilated left ventricle. AB - The operative mortality and morbidity in patients with severe left ventricular dysfunction who undergo coronary artery bypass grafting (CABG) remain high. The low ejection fraction is the major risk factor for operative mortality. However, ejection fraction (EF) alone may not necessarily be an accurate predictor of operative mortality. We studied the correlation between indices of left ventricular volume and operative mortality. One thousand patients undergoing isolated coronary bypass operations were divided into three groups according to their preoperative ejection fraction. Fifty patients (group I) had severe left ventricular dysfunction (EF < or = 0.3), 56 patients (group II) had moderately left ventricular dysfunction (0.3 < EF < or = 0.4) and 894 patients (group III) had good left ventricular function (EF > 0.4). We analyzed the relationship between hospital mortality and left ventricular volume in 106 patients with an EF < or = 0.4. RESULTS: Cardiac index was not significantly different among the three groups. The left ventricular end-diastolic pressure (LVEDP) and mean pulmonary artery pressure in groups I an II were higher than those in group III. The left ventricular end-diastolic volume (LVEDV) was 146 +/- 44 ml/m2 in Group I, 112 +/- 31 ml/m2 in Group II and 82 + 30 ml/m2 in Group III, respectively (Group I versus II, p < 0.05, Group I and II versus III, p < 0.01). The left ventricular end-systolic volume (LVESV) was 111 +/- 38 ml/m2 in Group I, 72 +/- 21 ml/m2 in Group II and 30 +/- 14 ml/m2 in Group III, respectively (Group I versus II, p < 0.05, Group I and II versus III, p < 0.01). The LVEDV and LVESV were higher in Group I than in Group II and both in Groups I and II were higher than in Group III. The hospital mortality of any cause before discharge was 8.0% (4/50) in Group I, 3.6% (2/56) in Group II, and 2.0% (18/894) in Group III. The mortality in Group I was higher than that in Group III, but the mortality between Groups I and II was not different. We assessed correlations between large left ventricle with left ventricular dysfunction and operative mortality in 106 patients with ejection fractions of < or = 0.4. The hospital mortality in patients with both under fraction 0.4 and an LVESV > or = 140 ml/m2 was 50% (4/8). This rate was higher than in patients with an LVESV between 80 and 140 ml/m2 (1.8%, 1/55) (p = 0.0006) and an LVESV less than 80 ml/m2 (2.3%, 1/43), (p = 0.0013). The hospital mortality in patients with an LVEDV > or = 200 ml/m2 was 67% (4/6). It was also higher than that in patients with an LVEDV between 200 and 120 ml/m2 (1.7%, 1/58), (p = 0.0001), and an LVEDV less than 120 ml/m2 (2.4%, 1/42), (p = 0.0004). We conclude that patients with a low ejection fraction and an elevated LVESV or LVEDV are at increased risk for hospital death following CABG. PMID- 9358811 TI - Percutaneous transhepatic gallbladder drainage for acute acalculous cholecystitis following cardiovascular surgery. AB - Four (1.2%) out of 321 patients required percutaneous transhepatic gallbladder drainage (PTGBD) following cardiovascular surgery. Cholecystitis was initially suspected based upon the occurrence of postoperative fever and the results of abdominal X-ray films. The main physical finding was tenderness of the right upper quadrant abdomen in all patients. Spontaneous pain and Blumberg's sign were not apparent. Distension of the gallbladder and sludge in the gall-bladder were detected in all four patients by ultrasonography, but calculi were not observed. Thickening and edema of the gallbladder wall, generally suggestive of cholecystitis, were observed in only one patient. PTGBD was performed from 5 to 43 (mean 16) days after surgery. The drained fluid was concentrated bile and not purulent. High fever dropped and serum transaminase and C-reactive protein levels decreased within three days after PTGBD. Bacteriologic examinations of the bile and arterial blood were negative in all cases. No complications as a result of PTGBD introduction occurred. PTGBD is a safe and effective procedure, and therefore should be actively performed even in the early phase of acute cholecystitis. PMID- 9358810 TI - Long-term clinical performance of Sorin tilting-disc mechanical prostheses in the mitral and aortic position. AB - OBJECTIVE: Analyzing the long term performance of sorin tilting-disc mechanical prostheses. EXPERIMENTAL DESIGN: Retrospective patient-oriented study. The total follow-up was 460.2 patient-years. Follow-up data was obtained from the patients themselves or from their relatives. SETTING: Department of Cardiovascular Surgery in a general community hospital. PATIENTS: Seventy four patients undergoing valve replacement with Sorin tilting-disc mechanical prostheses between May, 1982 and July 1991. INTERVENTIONS: Thirty one of those patients underwent isolated mitral valve replacement (MVR) and 43 isolated aortic valve replacement (AVR). MEASURES: The incidence of the different complications is expressed as linearized rates. Actuarial analysis was performed with the Kaplan-Meier method. RESULTS: Linearized rates for MVR and AVR for the different complications (events per 100 patient-years) were, respectively: Late mortality: 4.5 +/- 1.6 and 1.8 +/- 0.8; Thromboembolism: 3.4 +/- 1.4 and 1.1 +/- 0.6; Anticoagulant-related hemorrhage: 2.8 +/- 1.3 and 0.3 +/- 0.3; Prosthetic endocarditis: 1.1 +/- 0.8 and 0.7 +/- 0.5; Non-structural dysfunction: 0.5 +/- 0.5 and 1.1 +/- 0.6; Reoperation: 1.1 +/ 0.8 and 0.3 +/- 0.3. Actuarial probabilities of freedom from the different complications were, respectively, at 13 years follow-up for MVR and 12 years follow-up for AVR, the following: Late mortality: 45.7 +/- 12.4% and 70.3 +/- 7.9%; Thromboembolism: 74.6 +/- 10.8% and 90.7 +/- 5.1%; Anticoagulant-related hemorrhage: 79.4 +/- 11.6% and 97.3 +/- 2.7%; Prosthetic endocarditis: 92.7 +/- 4.9% and 91.2 +/- 6.4%; Non-structural dysfunction: 95.6 +/- 4.3% and 88.2 +/- 6.6%; Reoperation: 83.6 +/- 11.8% and 97.3 +/- 2.7%. All valve-related mortality and morbidity: 42.2 +/- 11.0% and 56.7 +/- 8.6%. There was no instances of prosthetic structural failure. CONCLUSIONS: The Sorin mechanical prosthesis presents a good durability and its performance in the long term is comparable to other tilting-disc devices of the same generation. PMID- 9358813 TI - Surgical repair of pulmonary stenosis with intact ventricular septum in a 68-year old woman. AB - Patients with mild pulmonary stenosis after infancy rarely have symptoms or develop increasing obstruction. We experienced a 68-year-old woman with severe pulmonary valvar and infundibular stenosis (peak to peak pressure gradient = 80 mmHg). She had been pointed out heart disease at the age of six. Endocarditis at the age of 17 might induce calcification of valve and affect the progression of pulmonary stenosis, and moreover, which might gradually develop severe subvalvar obstruction and poststenotic aneurysm of pulmonary trunk. She refused operative intervention because of mild clinical symptoms (NYHA class II), but we recommended surgical repair due to repeated transient ischemic attacks, which were suspected paradoxical embolism through persistent foramen ovale. She underwent pulmonary valvotomy and infundibular resection and is doing well. PMID- 9358812 TI - Concomitant cardiac and pulmonary operation. Pulmonary mechanics and outcome of phrenic nerve injury. AB - OBJECTIVE: We describe the postoperative respiratory failure due to the phrenic nerve injury in the setting of concomitant cardiac and pulmonary operation. EXPERIMENTAL STUDY: Prospective study. SETTING: Department of Cardiac and Thoracic Surgery Osaka University Medical School. PATIENTS AND INTERVENTIONS: From January 1984 to December 1993, 5 patients (1.4%) underwent the concomitant cardiac and pulmonary operation out of 359 patients who received surgical treatment for lung cancer at our institution. MEASURES AND RESULTS: Three (60%) out of 5 patients required prolonged mechanical ventilation despite the absence of cardiac complication, lung edema or pneumonia. Diaphragm function and work of breathing were measured in two patients before and after weaning from mechanical ventilation. Phrenic nerve dysfunction was consistent with the result that trans diaphragmatic pressure (delta Pdi) was low, a ratio of gastric to esophageal pressure swing (delta Pga/delta Pes) was abnormally negative, and work of breathing (WOB) was high. Phrenic nerve function restored associated with clinical improvement. CONCLUSIONS: Diaphragm dysfunction and an increase in work of breathing may be potential causes of respiratory failure in patients after concomitant cardiac and pulmonary operation. This compromise in respiratory mechanics should not be overlooked in the postoperative care, which may lead to the best management in postoperative respiratory care. PMID- 9358814 TI - Unifocalization and systemic-to-pulmonary shunt using internal mammary artery for tetralogy of Fallot and pulmonary atresia with diminutive pulmonary artery and arborization anomaly. AB - We report successful palliation of tetralogy of Fallot and pulmonary atresia with diminutive pulmonary arteries and arborization anomaly. Unifocalization and systemic-to-pulmonary artery shunt using an internal mammary artery were successfully performed. The internal mammary artery shunt provided a sufficient blood supply for a diminutive and fragile pulmonary artery, and brought a progressive improvement of exercise intolerance and polycythemia probably due to the growth potential. PMID- 9358815 TI - Ruptured aneurysm sinus of Valsalva and Gerbode defect with severe tricuspid and aortic regurgitation. A case report and its surgical correction. AB - An unusual early, childhood presentation in a case with reputured non-coronary sinus of Valsalva aneurysm with Gerbode defect and severe pulmonary hypertension is described. The reasons for early rupture are discussed and anatomically important relations of membranous septum, fibroskeleton of heart and conduction system are schematically elucidated. Associated severe tricuspid and aortic regurgitation are explained to be secondary effects following the rupture of aneurysm. A technique of surgical correction of this rare association of anomalies using single PTFE patch is illustrated, cautiously safeguarding the closely related conduction system. Regurgitant aortic and tricuspid valves were also successfully repaired. In retrospect, early repair before rupture of aneurysm and onset of severe pulmonary hypertension may be more beneficial, which would also prevent the leakage of semilunar and atrioventricular valves. PMID- 9358816 TI - The effect of ischemic conditioning on gastric wound healing in the rat: implications for esophageal replacement with stomach. AB - BACKGROUND: Esophagectomy, with gastric pull up replacement, is not uncommonly complicated by leakage from the esophagogastrostomy anastomosis. Occult ischemia of the mobilized gastric fundus is a major etiological factor for anastomotic leakage. Gastric tissue perfusion can be improved by ischemic conditioning ("delay" phenomenon). OBJECTIVE: To test the hypothesis that ischemic conditioning will improve gastric wound healing, and reduce the incidence of anastomotic dehiscence, in a rodent model of partial gastric devascularization. EXPERIMENTAL DESIGN: Laboratory study of gastric wound healing in rats. ANIMALS: Forty-five Sprague-Dawley rats (3 groups of 15 rats). INTERVENTIONS: All animals underwent laparotomy on day 0. Group 1 (control) and group 3 (acute ischemia) rats had sham laparotomies done. Group 2 (ischemic conditioning) rats underwent laparotomy and left gastric artery ligation. On postoperative day 14, all animals underwent repeat laparotomy; gastrotomy wounds were created and sutured. Group 1 (control) and group 2 (ischemic conditioning) rats had gastrotomy alone, while group 3 (acute ischemia) rats also underwent left gastric artery ligation. All rats were sacrificed 5 days after gastrotomy and wound healing was assessed. MEASURES: Gastrotomy wounds were assessed for dehiscence, bursting strength, and hydroxyproline concentration. RESULTS: Anastomotic dehiscence did not occur in group 1 (control) or group 2 (ischemic conditioning) rats. Four of 15 rats (27%) in group 3 (acute ischemia) suffered anastomotic dehiscence (p = 0.028). Wound bursting pressure in the three groups was not significantly different (group 1- 96.3 +/- 8.3 mmHg, group 2--91.1 +/- 4.8 mmHg, group 3--70.9 +/- 12.7 mmHg, p = 0.13). Wound hydroxyproline concentration in the control group was significantly higher than in the other 2 groups (group 1--0.124 +/- 0.005 mumol/mg, group 2- 0.113 +/- 0.007 mumol/mg, group 3--0.102 +/- 0.006 mumol/ mg, p = 0.04), but there was no difference between the acute ischemia and the ischemic conditioning groups (p = 0.24). CONCLUSIONS: In this rodent model of partial gastric devascularization, ischemic conditioning reduced the incidence of anastomotic dehiscence. Wound bursting strength and hydroxyproline concentration were not affected by ischemic conditioning. Therefore, the harmful effect of ischemia, and the beneficial effect of ischemic conditioning, are probably not primarily related to synthesis of wound collagen. Ischemic conditioning of the stomach is a concept that may prove clinically useful in reducing the incidence of leakage from esophagogastrostomy anastomoses. PMID- 9358817 TI - CT-guided transthoracic needle biopsy in the diagnosis of chest tumours. AB - Percutaneous Transthoracic Needle Biopsy (PTNB) is an accepted technique for the diagnosis of suspected intrathoracic malignancy and, although the appropriate indications have not been clearly defined yet, its use has rapidly expanded in the last years. The authors reviewed retrospectively their experience over one year time biopsies and analyzed some controversies regarding this diagnostic procedure. PMID- 9358818 TI - Continuous paravertebral block for post thoracotomy analgesia in children. AB - The use of continuous paravertebral analgesia was studied in 15 children with a mean age of 9.8 years (2-16 years). Nine patients received pre-emptive and postoperative paravertebral analgesia while six children studied earlier in the series received only post operative paravertebral analgesia. Excellent pain relief was attained in all patients, as assessed by the graded pictures of facial expression or visual analogue pain scores and morphine requirements. There were no pulmonary complications and no complications related to the continuous paravertebral infusion of bupivacaine. We conclude that continuous paravertebral block is an effective and safe method for post thoracotomy pain relief in children. PMID- 9358819 TI - Radical excision of invasive thymoma with intracaval and intracardial extension: a successful case report. AB - A case of invasive thymoma with intracaval and intracardiac extension is reported. The use of cardiopulmonary bypass was necessary to achieve a radical excision of the tumor thus avoiding early death due to cardiovascular complications. This highly unusual mode of tumor presentation makes this particular case worth reporting. PMID- 9358820 TI - The secrets of the unknown trauma patient. The hazards of undiagnosed subclavian steal syndrome in the chest trauma patient: a case report. AB - Chest trauma can be complicated, among others, with cardiac tamponade. This life threatening condition should be treated promptly and adequately to assure a positive outcome. During the rapid events that take place in the emergency room with the arrival of a polytrauma patient, anamnestic data are not always available, especially if dealing with a non-cooperative, unaccompained traumatized patient. The following case report describes our experience with a chest trauma patient after a vehicle accident, who was admitted to our ward exhibiting a constellation of signs compatible with a cardiac tamponade. The only demonstrable objective signs included distended mediastinum and heart shadow on the chest X ray and muffled heart sounds. However, despite the impressive clinical picture, the patient continued to exhibit constant, though low blood pressure measurements and after a short period of observation, given the homodynamic stability, it was decided against pericardiocenthesis. The "secret" of our patient was finally discovered at angiography, when a left subclavian steal was diagnosed. The literature in this matter is discussed, stressing the importance of anamnestic data in the trauma patient. Most importantly, we address the significance of relying on hard clinical data (homodynamic stability) rather than on isolated signs (widened mediastinum/heart shadow) to reach as accurate a diagnosis as possible before pursuing invasive, usually not-innocuous procedures (pericardiocenthesis). PMID- 9358821 TI - Ischemic conditioning of the rat stomach is not mediated by endogenous basic fibroblast growth factor. PMID- 9358822 TI - Plasmapheresis is not justified in treatment of rhabdomyolysis and acute renal failure. PMID- 9358823 TI - Increase in HIV and tobacco-related cancers expected. PMID- 9358824 TI - Geriatric medicine in South Africa--the onus is on medical schools. PMID- 9358825 TI - Compensatory functions and therapies in the elderly--everything is not lost. PMID- 9358827 TI - A new model for the pathophysiology of Alzheimer's disease. Aluminium toxicity is exacerbated by hydrogen peroxide and attenuated by an amyloid protein fragment and melatonin. AB - OBJECTIVES: Although Alzheimer's disease (AD) is the leading cause of dementia in developed countries, there is an as yet unexplained lower prevalence of the disease in parts of Africa. AD is characterised by a catastrophic loss of neurons; free radicals (oxidative toxins) have been implicated in the destruction of the cells through the process of lipid peroxidative damage of cell membranes. Previously aluminium (Al) and a fragment of beta amyloid (A beta 25-35) were shown to exacerbate free-radical damage, while melatonin reduced this effect. The aim of the present study was: (i) to investigate the conditions determining the toxicity of Al and A beta 25-35; and (ii) to assess whether melatonin could attenuate the damage done by both aluminium and the amyloid fragment, thus suggesting a pathway for the aetiology of AD. DESIGN: An in vitro model system was used in which free radicals were generated, causing lipid peroxidation of platelet membranes, thus simulating the disease process found in the brain. RESULTS: 1. Al and A beta 25-35 caused lipid peroxidation in the presence of the iron (II) ion (Pe2+), Al being more toxic than A beta 25-35. 2. A beta 25-35 attenuated the lipid peroxidation promoted by Al. 3. Hydrogen peroxide (H2O2) greatly exacerbated the toxicity of Al and A beta 25-35. 4. Melatonin prevented lipid peroxidation by Al and A beta 25-35 in the absence of H2O2, but only reduced the process when H2O2 was present. CONCLUSIONS: In the light of the results obtained from the present study, the following hypotheses are formulated. 1. In AD, excessive quantities of Al are taken up into the brain, where the Al exacerbates iron-induced lipid peroxidation in the lysosomes. 2. In response, the normal synthetic pathway of amyloid protein is altered to produce A beta fragments which attenuate the toxicity of Al. In the process of sequestering the Al and iron, immature plaques are formed in the brain. 3. Microglia are activated, in an attempt to destroy the plaques by secreting reactive oxygen species such as H2O2. At this point in the disease process, lipid peroxidation causes a catastrophic loss of brain cells. 4. Melatonin, together with other free radical scavengers in the brain, reduces the free-radical damage caused by Al and A beta, except in the latter stages of the disease process. Since melatonin is produced by the pineal gland only in the dark, the excess of electric light in developed countries may help explain why AD is more prevalent in these countries than in rural Africa. PMID- 9358828 TI - Zinc and platelet membrane microviscosity in Alzheimer's disease. The in vivo effect of zinc on platelet membranes and cognition. AB - OBJECTIVES: To investigate the effects of oral zinc supplementation on: (i) plasma zinc concentrations; (ii) platelet membrane microviscosity in vivo; and (iii) cognitive function of Alzheimer's disease (AD) patients. DESIGN: An open labelled pilot study. SETTING: University of Stellenbosch Medical School and Stikland Hospital. SUBJECTS: Six volunteer AD patients. OUTCOME MEASURES: Plasma zinc levels, platelet membrane microviscosity and cognition (MMSE and ADAS-cog tests). RESULTS: Oral zinc supplementation (30 mg/day) did not increase plasma zinc levels significantly, but significantly increased platelet membrane microviscosity (P = 0.02; 6 patients). Four patients, who underwent 12 months of evaluation, showed modest cognitive improvement on psychometric testing (Mini Mental State Examination and the cognitive portion of the Alzheimer's Disease Assessment scale scores). CONCLUSIONS: While earlier literature promoted the use of zinc in AD patients, a recent study has contradicted this and implicated zinc in the aetiology of Alzheimer's disease. On the basis of the above results, it may be premature to single out zinc as a causal agent in AD. PMID- 9358829 TI - Thyroid dysfunction in the elderly. AB - OBJECTIVES: To determine the prevalence of thyroid dysfunction in institutionalised elderly people in Cape Town and to assess the usefulness of an abnormal thyroid-stimulating hormone (TSH) concentration as a screening test in this group. DESIGN: Cross-sectional survey. SETTING: Four old-age homes in Cape Town. SUBJECTS: Old-age home residents aged 60 years and over. OUTCOME MEASURES: Serum concentrations of TSH, free thyroxine and free tri-iodothyronine. RESULTS: Serum TSH estimations were performed on 658 participants, and were abnormal in 103 (15.6%)-41 (6.2%) being elevated (> 5.0 microU/ml) and 62 (9.4%) being low (> 0.4 microU/ml). There were 3 newly diagnosed cases of hyperthyroidism and 7 of hypothyroidism. Subclinical disease was diagnosed in 40 subjects. The overall prevalence of thyroid dysfunction in this population was 11.2%. In 22 (3.4%) this had previously been recognised, while in 50 (7.8%) the dysfunction was newly diagnosed by the current survey. The positive predictive value of a TSH concentration > 20 microU/ml in predicting hypothyroidism is 67%, while it will predict 100% of cases of subclinical hypothyroidism. A TSH concentration < 0.1 microU/ml will predict 23% of cases of hyperthyroidism, but 81% of cases of subclinical disease. CONCLUSIONS: The prevalence of thyroid dysfunction in institutionalised elderly people in Cape Town is similar to that reported for elderly people in other centres. Thyroid dysfunction had not previously been recognised in approximately two-thirds of the subjects in this study. The serum TSH concentration is a reliable screening test for thyroid dysfunction in the elderly, but is less useful if used to identify biochemical thyroid disease. An elevated TSH concentration is a better predictor of thyroid dysfunction in the elderly than a depressed TSH concentration. PMID- 9358830 TI - Physical activity, change in blood pressure and predictors of mortality in older South Africans--a 2-year follow-up study. AB - OBJECTIVE: A 2-year follow-up study of a cohort of 200 historically disadvantaged older South Africans was conducted to: (i) characterise current levels of habitual physical activity; (ii) relate physical activity to current risk factors for chronic disease; and (iii) identify risk factors associated with 2-year mortality. The baseline sample, drawn in 1993, was found to have a high prevalence of hypertension (71.7%). RESEARCH DESIGN: Retrospective cohort study. METHODS: A baseline sample of 200 persons aged > or = 65 years, resident in the Cape Peninsula, was randomly drawn by means of a two-stage cluster design. Baseline measurements included: anthropometry, waist/hip ratio, systolic and diastolic blood pressure, body mass index (BMI), serum albumin, serum ferritin, haemoglobin and fasting plasma glucose levels, plasma lipid profiles, oral glucose tolerance test and self-reported health status. Subjects were revisited after 2 years, at which time an adapted version of the Yale Physical Activity Survey was administered and measurements of blood pressure and anthropometry were repeated. STATISTICAL ANALYSES: Spearman's rank-order correlations were used to describe relationships between various current risk factors and physical activity. Logistic regression was used to determine predictors of 2-year mortality from baseline data. RESULTS: At follow-up, 142 of the subjects (66 men, 76 women) were traced and measurements collected. Thirty-two subjects were reported to have died by relatives living in the same household (22 men, 10 women). Levels of reported physical activity in the survivors were two-thirds lower than those reported in a sample of North Americans of similar age. There was an inverse association between age and physical activity (r = -0.31; P < 0.0005) and a positive association between BMI and physical activity (r = 0.29; P < 0.005). There was, however, no association between physical activity and systolic or diastolic blood pressure. In men, BMI in the lower tertile (P = 0.07) and serum ferritin levels were positively associated with increased mortality. Serum albumin levels were protective over the 2-year follow-up period (OR = 0.85; P < 0.05). In women, being diabetic (OR = 4.88; P = 0.06) and having a waist/hip ratio in the upper tertile (OR = 3.26; P = 0.06) were associated with mortality. CONCLUSIONS: Physical activity levels in this sample of older historically disadvantaged South Africans were habitually low. Simple anthropometric assessments incorporating weight and waist/hip ratio, together with serum albumin measurements, may be useful to screen general health risk for older adults at primary care level and provide indications for social or medical intervention. Further, strategies for earlier detection and effective management of diabetes, particularly in older women, may reduce premature mortality in this population. PMID- 9358831 TI - The role of chair exercises for older adults following hip fracture. AB - OBJECTIVES: To examine the influence of regular participation in chair exercises on postoperative deconditioning in respect of selected physiological, psychological and anthropometric variables. DESIGN: Quasi-experimental, non randomised control group pre-test/post-test design where test group (N = 20) and control group (N = 10) were not equivalent because random selection and assignment were not possible. SUBJECTS: Patients discharged from an orthopaedic ward 8-10 days after hip surgery who were cognitively competent (mean score on Mini-Mental State Examination 26 (SD 3.5)), sedentary (mean score on Habitual Physical Activity Questionnaire for the Elderly 7.4 (SD 3.3)) women aged 70 years and above (mean 80 (SD 6.6) years). SETTING: Hip fracture patients admitted to a multidisciplinary geriatric hospital for further medical observation. MEASUREMENTS: Abstraction of medical records provided information about co morbidities and questionnaires assessed demographic, affective and cognitive function. Physiological, psychological and anthropometric status was measured pre and post-intervention. RESULTS: Data revealed high variability, suggesting that the effect of the independent variable was obscured by the heterogeneity of the cohort. Both groups improved similarly in grip strength, and in levels of depression and confidence. Body composition data explained the weight maintenance as a consequence of significant gains in fat-free mass in the experimental group. The significant change in systolic blood pressure and heart rate over the exercise and recovery period suggested that the 6-week period of moderate intensity exercise was adequate to stimulate cardiovascular adaptations. CONCLUSIONS: Whether or not the submaximal chair exercise regimen was of optimal benefit remains unclear. However, the intervention did appear to have contributed to the maintenance of the physical condition of older women temporarily disabled as a result of a fracture and subsequent hip surgery. PMID- 9358832 TI - Hoarding symptoms in patients on a geriatric psychiatry inpatient unit. AB - BACKGROUND: While collecting may be a normal behaviour, hoarding is a symptom of various psychiatric disorders, including obsessive-compulsive disorder (OCD) and obsessive-compulsive personality disorder (OCPD). Although anecdotal reports suggest that hoarding is not uncommon in geriatric psychiatry populations, its psychopathological correlates in such samples have not been well characterised. METHODS: The presence of clinically significant hoarding symptoms was screened for in 100 consecutive patients in a geriatric psychiatry inpatient unit. Both patient and collateral histories were obtained. When hoarding symptoms were present, a detailed history of their phenomenology was obtained by means of a structured questionnaire and the response of hoarding symptoms to treatment during hospitalisation was monitored. RESULTS: Clinically significant hoarding was found in 5/100 subjects. Four of these 5 patients met DSM-IV criteria for schizophrenia (paranoid subtype), with onset of symptoms coinciding with increased symptoms of dementia. The fifth patient met criteria for bipolar disorder (manic episode), also had symptoms of dementia, and had a lifelong history of hoarding. Hoarding behaviours responded to antipsychotic treatment in 3 of the 5 patients. CONCLUSIONS: A history of hoarding may be useful in many psychiatric patients, but psychopathological correlates of this symptom are likely to vary with age. In a geriatric psychiatry inpatient population hoarding was associated not with OCD or OCPD, but rather with paranoid schizophrenia and increasing symptoms of dementia. Dopamine blockers appeared useful in decreasing hoarding in some patients, raising interesting questions about the neurobiology of this symptom. PMID- 9358833 TI - Are professional interpreters needed in the South African health care services? PMID- 9358836 TI - Kernicterus associated with elevated predominantly direct-reacting bilirubin. PMID- 9358837 TI - Parodoxical rise in urinary albumin levels after treatment of essential hypertension. PMID- 9358839 TI - Lack of evidence for early cases of HIV seropositivity/AIDS in the Free State region. PMID- 9358840 TI - Expressive dysphasia presenting as psychiatric disorder--a case report. PMID- 9358842 TI - Minimally invasive, maximally morbid. PMID- 9358841 TI - A difficult decision. PMID- 9358843 TI - Intracytoplasmic sperm injection--new hope for severe male factor infertility. PMID- 9358844 TI - Selective intra-operative internal jugular venous sampling for rapid immunoradiometric assay of intact parathyroid hormone during parathyroid surgery. PMID- 9358847 TI - Tyramine amplification technique in routine immunohistochemistry. AB - Signal amplification in immunohistochemistry via binding of biotinylated tyramine to proteins near the site of peroxidase-labeled antibodies is a promising new technique, but studies investigating a wide range of markers are lacking. The tyramine amplification technique (TAT) was investigated on 85 antibodies using a simple and fast protocol, and TAT results were compared to those obtained with conventional immunohistochemistry. Using TAT, most of the markers could be 5- to 50-fold further diluted and still showed identical staining results compared with standard stainings (maximal 500-fold). However, the variable reactivity of the different markers with TAT underlines the need for individual testing of every antibody to determine the optimal dilution. Some antibodies against cell adhesion molecules could be demonstrated for the first time in archival, formalin-fixed tissue sections. TAT, if carefully evaluated, offers a revolutionary improvement for modern immunostaining, either to increase sensitivity or primary antibody dilutions (cost reduction). From a methodological point of view, immunohistochemistry has not reached its limits by far and TAT is an important progressive step in this developmental process. PMID- 9358846 TI - Biotinyl-tyramide: a novel approach for electron microscopic immunocytochemistry. AB - The biotinyl-tyramide protocol recently introduced for sensitive light microscopic immunocytochemistry was applied to electron microscopy and revealed various tissue antigens with high resolution. The protocol consists of an indirect method in which thin tissue sections are incubated successively within a specific primary antibody, followed by a biotinylated secondary antibody, streptavidin-HRP, and then finally with biotinyl-tyramide. The reaction product appears as a dense filamentous material that is deposited over particular cellular compartments. The labeling obtained for the antigens tested, amylase and heat-shock protein 70 in pancreatic acinar cells, insulin in pancreatic beta cells, and carbamoyl phosphate synthetase and catalase in liver tissue, was found to be highly specific, with the labeling for each antigen confined to its particular cellular compartment. Background levels and nonspecific deposition of the staining were negligible. The use of biotinyl-tyramide therefore appears to be an alternative sensitive technique for immunoelectron microscopy. PMID- 9358848 TI - Receptors for the chemoattractants C5a and IL-8 are clustered on the surface of human neutrophils. AB - We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a "nonactivated" state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL 8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an "activated" state, by mixing the blood with N-formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration. PMID- 9358849 TI - Type IIA procollagen amino propeptide is localized in human embryonic tissues. AB - Type II procollagen is synthesized in two forms generated by the alternative splicing of its precursor mRNA. The alternatively spliced domain, exon 2, encodes the 69-amino-acid cysteine-rich region of the NH2 propeptide. Studies of mRNA expression have shown that the longer form, designated Type IIA procollagen, is synthesized by chondroprogenitor cells and various noncartilaginous tissues. The shorter form, Type IIB procollagen, is synthesized by differentiated chondrocytes. As the initial step in our investigations of the function of the Type IIA procollagen, the protein domain corresponding to exon 2 was created as a recombinant fusion protein and used to raise antibodies in rabbits. The resulting antiserum was specific for Type IIA procollagen NH2 propeptide as shown by ELISA, Western blotting, and immunofluorescent co-localization with the triple-helical domain of Type II collagen. Type IIA procollagen was identified in tissue culture medium of 54-day human fetal ribs. Confocal microscopy was used to localize the Type IIA NH2 propeptide in Day 50 and 53 human embryos. In the digital rays of the developing hand, where only Type IIA procollagen mRNA was detected, Type IIA procollagen NH2 propeptide was observed in the extracellular matrix. The presence of Type IIA procollagen NH2 propeptide was observed in the cartilage of the developing long bones of the lower arm and vertebral bodies even though these tissues synthesize Type IIB mRNA at this developmental stage. Type IIA procollagen NH2 propeptide was localized in the developing trachea, a cartilage that does not undergo endochondral bone formation. Type IIA NH2 propeptide was also localized in noncartilaginous tissues known to synthesize Type IIA mRNA, such as the intervertebral area, perichondrium, notochordal sheath, and neuroepithelium of the otic vesicle. In most tissues, co-localization with antiserum against the triple-helical domain of Type II collagen was observed. Positive immunoreactivity with the Type IIA NH2 propeptide antiserum indicates, for the first time, that this propeptide is present in the tissue. Co localization of NH2 propeptide antibodies with the triple-helical domain of the collagen molecule suggests that Type IIA procollagen is intact in the extracellular matrix of these tissues. Taken together, these results strongly suggest that around cells that synthesize Type IIA procollagen mRNA, Type IIA procollagen NH2 propeptide is secreted and deposited into the extracellular matrix. In light of these results, we predict that Type IIA procollagen plays a role in differentiation of tissues that augments its purely architectural function. PMID- 9358850 TI - Immunolocalization of CD44 and the ezrin-radixin-moesin (ERM) family in the stratum intermedium and papillary layer of the mouse enamel organ. AB - We studied the immunohistochemical localization of CD44 and the ezrin-radixin moesin (ERM) family of actin binding proteins in mouse enamel organ, using confocal laser scanning microscopy and transmission electron microscopy to clarify their role in cytoskeletal organization. At the differentiation stage of ameloblasts, immunoreactivity to CD44 was detected on the plasma membrane of the inner enamel epithelium, the stellate reticulum, the stratum intermedium, and the external enamel epithelium. In accordance with the differentiation of preameloblasts into secretory ameloblasts, immunoreactivity increased in the stratum intermedium cells. At the maturation stage, intense immunoreactivity was observed on the papillary layer cells. For the ERM family, the stratum intermedium and the papillary layer cells were stained with anti-ezrin and radixin monoclonal antibodies but not with the anti-moesin antibody. Electron microscopic observations revealed that CD44, ezrin, and radixin were localized in the region at which preameloblasts came into contact with the stratum intermedium at the differentiation stage. At the secretory and maturation stages, they were concentrated in the microvilli of the stratum intermedium and the papillary layer cells. These findings suggest that the CD44-ezrin-radixin-actin filament system is involved in cell-cell interaction between preameloblasts and the stratum intermedium, and in the cytoskeletal organization of the cells in the stratum intermedium and the papillary layer. PMID- 9358852 TI - Autometallographic silver enhancement of zinc sulfide crystals created in cryostat sections from human brain biopsies: a new technique that makes it feasible to demonstrate zinc ions in tissue sections from biopsies and early autopsy material. AB - We present a new technique that allows zinc ions in synaptic and secretory vesicles of biopsy and early autopsy material (< 2 hr post mortem) to be transformed to nanometer-sized zinc sulfide crystal lattices for subsequent autometallographic (AMG) development. Human brain biopsies, or other tissue samples containing zinc-enriched (ZEN) cells, are frozen in liquid nitrogen or by CO2 gas immediately after removal. The tissue blocks are cut in a cryostat and the sections placed on glass slides. The slides are transferred to an H2S exposure chamber placed in a -15 C freezer. After 1-24 hr of gas exposure the sections are removed from the chamber, fixed while thawing, and dehydrated. The sections are then exposed to an AMG developer. AMG causes silver enhancement of zinc sulfide crystal lattices created in the tissues through the H2S exposure, making them visible. It is imperative that the tissues are frozen instantaneously after removal, because loosely bound or free zinc ions start leaving their vesicular compartment soon after death. The AMG technique can, despite inadequate fixation and damage to the tissue caused by freezing, also be used to trace zinc ions at ultrastructural levels, and it is demonstrated that zinc ions in the human neocortex are located in synaptic vesicles. In the few human biopsies analyzed thus far, the light microscopic pattern created by the silver-enhanced ZEN terminals resembles that seen in the neocortex of rat brain. The technique has been applied to cryostat sections from neocortex biopsies of five individuals undergoing brain surgery. Biopsies from three patients resulted in satisfactory AMG-stained sections. Rat brains removed and frozen immediately after decapitation constituted the material on which the present technique was developed. Such material results in an almost uniform high quality of staining, and we found that unexposed sections can be stored for at least 5 months at -80 C without ensuing significant loss of AMG staining intensity. PMID- 9358851 TI - Copper-metallothionein in the kidney of macular mice: a model for Menkes disease. AB - Menkes disease is an X-linked disorder of copper metabolism. Excess amounts of copper in the kidney of Macular mice, a model for this disease, were found as copper-metallothionein (Cu-MT) from kidney of the mice. Histochemical studies of Cu-MT based on its autofluorescent emission properties showed that the protein was predominant in the proximal convoluted tubule (PCT) cells of the cortex. PCT cells are known to be the primary site of the nephrotoxicity caused by heavy metals. MT mRNA was also observed in the cortex, indicating that the protein was biosynthesized in this region. On the basis of these results, we suggest that biosynthesis and degradation of Cu-MT occur repeatedly in the PCT cells of the cortex. We also compared the histochemical localization of Cu-MT in Macular mice and Long-Evans cinnamon rats, a model for Wilson's disease. The significance of this comparison is discussed. PMID- 9358853 TI - Localization of B-DNA and Z-DNA in terminally differentiating fiber cells in the adult lens. AB - We examined histochemically and immunohistochemically the distribution of B- and Z-DNA in the epithelium and terminally differentiating dog lens fiber cells. On the basis of anti-DNA antibody reactivity, qualitative and quantitative data on B and Z-DNA in cells were determined. Anti-B-DNA immunoreactivity gradually declined throughout nucleated fibers, with a precipitous decrease at approximately 90 microns. Anti-Z-DNA antibody binding decreased with a sudden loss of immunoreactivity at approximately 90 microns. The pattern of anti-B- and Z-DNA staining correlates with the loss of alpha-crystallin immunoreactivity, the major lens crystallin, and decreased eosin staining of proteins. Germinative zone cell nuclei showed the highest DNA probe binding values, followed by the superficial fibers, central zone, middle fibers, and deep fibers. The presence of single-stranded (ss)DNA in deeper fibers was detected by anti-ss-DNA antibodies. This is indicative of DNA degradation. These observations suggest that a dramatic reorganization of lens fiber cells' supramolecular order occurs at approximately 90 microns, the phase transition zone. PMID- 9358854 TI - Changes in morphology and spatial position of coiled bodies during NGF-induced neuronal differentiation of PC12 cells. AB - Interphase nuclei are organized into structural and functional domains. The coiled body, a nuclear organelle of unknown function, exhibits cell type-specific changes in number and morphology. Its association with nucleoli and with small nuclear ribonucleo-proteins (snRNPs) indicates that it functions in RNA processing. In cycling cells, coiled bodies are round structures not associated with nucleoli. In contrast, in neurons, they frequently present as nucleolar "caps." To test the hypothesis that neuronal differentiation is accompanied by changes in the spatial association of coiled bodies with nucleoli and in their morphology, PC12 cells were differentiated into a neuronal phenotype with nerve growth factor (NGF) and coiled bodies detected by immunocytochemical localization of p80-coilin and snRNPs. The fraction of cells that showed coiled bodies as nucleolar caps increased from 1.6 +/- 0.9% (mean +/- SEM) in controls to 16.5 +/- 1.6% in NGF-differentiated cultures. The fraction of cells with ring-like coiled bodies increased from 17.2 +/- 5.0% in controls to 57.8 +/- 4.4% in differentiated cells. This was accompanied by a decrease, from 81.2 +/- 5.7% to 25.7 +/- 3.1%, in the fraction of cells with small, round coiled bodies. SnRNPs remained associated with typical coiled bodies and with ring-like coiled bodies during NGF-induced recruitment of snRNPs to the nuclear periphery. Together with the observation that coiled bodies are also present as nucleolar caps in sensory neurons, the results indicate that coiled bodies alter their morphology and increase their association with nucleoli during NGF-induced neuronal differentiation. PMID- 9358855 TI - Retrovirus-mediated gene transfer into rat salivary gland cells in vitro and in vivo. AB - A retroviral vector DAP that encodes the human placental alkaline phosphatase (PLAP) and the neomycin-resistant gene was used to transduce the salivary gland derived cell line A5 in vitro and acinar cells in rat submandibular gland in vivo. Expression of the transduced PLAP gene was established by histochemical staining for heat-resistant AP and by determination of enzyme activity. From the in vitro experiments, we concluded that the salivary gland-derived cell line A5 can be infected by the retroviral vector DAP. In the transduced cells the viral long terminal repeat (LTR) promoter was effective, and the cells expressed heat stable PLAP which was localized mostly in the plasma membrane and could be released by treatment with bromelain or phosphatidyinositol-specific phospholipase C. A5-DAP cells secreted PLAP into the medium. Clones of A5-DAP cells expressed various levels of the enzyme. The level of enzyme activity in different clones was unrelated to growth rate. Retrograde ductal injection of the viral vector into the duct of the submandibular gland of rats resulted in integration and long-term expression of PLAP gene in acinar cells. Expression of PLAP was seen up to 25 days, the limit of the observation period. To facilitate integration of the viral DNA, cell division of acinar cells was induced by administration of the beta-adrenergic agonist isoproterenol before administration of the virus. PLAP was secreted into submandibular saliva. The data support the notion that salivary glands are suitable targets for gene transfer in vivo by a retroviral vector. PMID- 9358856 TI - Mucins (MUC1 and MUC3) of gastrointestinal and breast epithelia reveal different and heterogeneous tumor-associated aberrations in glycosylation. AB - In a comprehensive study, we examined the expression of the membrane and secretory mucins MUC1 and MUC3, respectively, in normal and neoplastic gastrointestinal and breast epithelia before and after specific alterations of their glycan structures by neuraminidase, alpha-fucosidase, or carbohydrate specific periodate oxidation. MUC1 mRNA was also identified in normal colorectal tissues by in situ hybridization. The data revealed that normal colorectal epithelia express both MUC1 mRNA and protein, which were detectable after periodate oxidation with all tested MUC1-specific antibodies. During tumorigenesis in the colon, MUC1 became recognizable without periodate treatment concomitantly with highly dysplastic lesions and the malignant state. In the breast, in which MUC1 is detectable with most antibodies in normal epithelium as well as in carcinomas, staining could be enhanced by pretreatment with periodate and casually by enzyme treatments. MUC3 was detectable in normal and neoplastic colorectal tissues and was more intensely stained after periodate oxidation. It was absent in normal breast even after pretreatment but was expressed in seven of 20 breast carcinomas. Therefore, incomplete glycosylation, abnormal distribution, and ectopic expression of mucins are characteristics of malignancy. Periodate oxidation may be widely applicable to immunohistochemistry for examining changes in glycosylation and for detecting antigens masked by glycans. PMID- 9358857 TI - Application of photoshop-based image analysis to quantification of hormone receptor expression in breast cancer. AB - The benefit of quantifying estrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer is well established. However, in routine breast cancer diagnosis, receptor expression is often quantified in arbitrary scores with high inter- and intraobserver variability. In this study we tested the validity of an image analysis system employing inexpensive, commercially available computer software on a personal computer. In a series of 28 invasive ductal breast cancers, immunohistochemical determinations of ER and PR were performed, along with biochemical analyses on fresh tumor homogenates, by the dextran-coated charcoal technique (DCC) and by enzyme immunoassay (EIA). From each immunohistochemical slide, three representative tumor fields (x20 objective) were captured and digitized with a Macintosh personal computer. Using the tools of Photoshop software, optical density plots of tumor cell nuclei were generated and, after background subtraction, were used as an index of immunostaining intensity. This immunostaining index showed a strong semilogarithmic correlation with biochemical receptor assessments of ER (DCC, r = 0.70, p < 0.001; EIA, r = 0.76, p < 0.001) and even better of PR (DCC, r = 0.86; p < 0.01; EIA, r = 0.80, p < 0.001). A strong linear correlation of ER and PR quantification was also seen between DCC and EIA techniques (ER, r = 0.62, p < 0.001; PR, r = 0.92, p < 0.001). This study demonstrates that a simple, inexpensive, commercially available software program can be accurately applied to the quantification of immunohistochemical hormone receptor studies. PMID- 9358859 TI - Spontaneous emission of phosphane from animal slurry treatment processing. AB - The degree of the emission of the mutagenic phosphane from animal slurry have become an issue of hygienic which so far has not been investigated. This work clearly detects spontaneously emitted free phosphane from animal slurry for the first time and correlates the degree of its emission with different disposal technologies. The subjects of the investigations are the simple storage process and processes involving biogas plants for the digestion both of pig and cattle slurry. Pig slurry generates about one magnitude more phosphane than cattle slurry. The maximum concentration detected in putrefaction gas was 14621 ppt(v/v). Putrefaction gas samples from fresh fecal slurry in primary (mediary storage) tanks followed by storage basins and sedimenters contains the highest concentrations. The mainly methanogenic biogas process generates the smallest concentrations but the highest fluxes of phosphane. Fluxes and concentrations in open basins are significantly higher during summer than in winter. The correlation of phosphane and dimethyldisulfide concentrations indicates that primary lytic processes play a role in the liberation of phosphane. Individual samples of the emission in air give a maximum value of 35 ppt. By comparison, measurement of phosphane in Hungarian digester gas from municipal sewage treatment show that maximum concentrations could be some orders of magnitude higher. Therefore the data base for phosphane from animal slurry must in future be expanded. The results of analysis so far achieved cannot be interpreted from either a human or veterinary medical viewpoint. PMID- 9358858 TI - Production of a monoclonal antibody by in vitro immunization that recognizes a native chondroitin sulfate epitope in the embryonic chick limb and heart. AB - We report the production of a monoclonal antibody (d1C4) by in vitro immunization that has immunoreactivity with a native chondroitin sulfate epitope in embryonic chick limb and heart. Murine lymphocytes were stimulated by direct exposure to unfixed, unsolubilized precartilage mesenchymal aggregates in high-density micromass culture derived from Stage 22-23 chick limb buds. Specificity of d1C4 reactivity was demonstrated by sensitivity of immunohistochemical staining to pretreatment with chondroitinase ABC or AC, preferential immunoreactivity with chondroitin-6-sulfate glycosaminoglycan (CS-C GAG) in ELISA, and competition of immunohistochemical staining with CS-C GAG. Immunohistochemical analysis of the expression of the d1C4 epitope revealed a striking localization of immunoreactivity in the extracellular matrix (ECM) of precartilage aggregates of chick limb mesenchyme in high-density micromass culture by 16 hr and the prechondrogenic limb core at Stage 23 in vivo. Immunoreactivity in both cultured limb mesenchyme and the embryonic limb continued through differentiation of prechondrogenic condensations into cartilage tissue. In the developing chick heart, d1C4 staining was found throughout the ECM of atrioventricular cushion tissue by Stage 25, but was localized to mesenchyme adjacent to the myocardium in the outflow tract cushions. There was an abrupt demarcation between d1C4-reactive intracardiac mesenchyme and unreactive extracardiac mesenchyme of the dorsal mesocardium in the Stage 22 embryo. This study demonstrates the efficacy of in vitro immunization of lymphocytes for the production of MAbs to native ECM constituents, such as CS-GAGs. Immunohistochemical data utilizing d1C4 suggest that CS-GAGs bearing this epitope may be important in early morphogenetic events leading to cartilage differentiation in the limb and valvuloseptal morphogenesis in the heart. PMID- 9358860 TI - Aspects of the central nervous drive to growth hormone secretion during aging. PMID- 9358861 TI - Hyperactivity of the hypothalamic-pituitary-adrenal axis in aging: a gerogenetic or an adaptive factor? PMID- 9358862 TI - Melatonin and aging: facts and artifacts. PMID- 9358863 TI - Neurosteroids: a role in aging? New functions in the central and peripheral nervous systems. PMID- 9358864 TI - Estrogens as neuromodulators. PMID- 9358865 TI - Neuroendocrine responses to exercise in healthy elderly humans and aging rats. PMID- 9358866 TI - Peptides, memory, food intake and aging. PMID- 9358867 TI - Altered pulsatile and coordinate secretion of pituitary hormones in aging: evidence of feedback disruption. AB - A novel thesis is that healthy aging of neuroendocrine axes is marked by disruption of orderly patterns of hormone release. This can be quantified by a recently validated approximate entropy statistic applied to 24-hour hormone profiles, e.g., luteinizing hormone (LH), and growth hormone (GH). Moreover, more subtle disturbances of feedback control are indicated by decreased conditional regularity or synchrony (higher cross-approximate entropy) within coupled axes, such as ACTH-cortisol, LH-testosterone, etc. Such alterations can precede any changes in mean serum hormone concentrations, thus highlighting an impact of age on the feedback control mechanisms that coordinate the flow of signaling information within a neuroendocrine axis or network. PMID- 9358868 TI - Neuroendocrine circadian rhythms in aging. PMID- 9358869 TI - The aging immune system: subsets, signals, and survival. PMID- 9358870 TI - Neuroendocrine and immune responses to stress in aging. PMID- 9358871 TI - Neurochemical, immunological and pharmacological assessments in a transgenic mouse model of the endocrine changes in depression. PMID- 9358872 TI - Zinc, nervous system and aging. PMID- 9358873 TI - Stress, inflammation and natural immunity in the aging process: a new theory. PMID- 9358874 TI - Variability of natural killer (NK) cell immune function in normal aging and senile dementia: pathophysiological implications. PMID- 9358875 TI - Immune aspects in elderly depression: peripheral blood mononuclear cell responses to mitogen stimulation and cytokine plasma concentrations. PMID- 9358876 TI - ADH and oxytocin in age-related minor cognitive impairment. PMID- 9358877 TI - The neuropathogenesis of delirium. PMID- 9358878 TI - The HPA-system in late-life depression. PMID- 9358879 TI - Problems of eating and appetite control in the elderly. PMID- 9358880 TI - Effects of aging on energy balance--intake and expenditure. PMID- 9358881 TI - Dietary patterns and cognitive functions in elderly subjects. PMID- 9358882 TI - Neuropeptides vasopressin (AVP), oxytocin (OXT) and corticotropin-releasing hormone (CRH) in the human hypothalamus: activity changes in aging, Alzheimer's disease and depression. PMID- 9358884 TI - Biochemical and cerebral morphometric correlates of physiological aging and senile dementia. PMID- 9358883 TI - Oxidative stress and dementia: new perspectives in AD pathogenesis. PMID- 9358885 TI - Peripheral markers of Alzheimer's disease. PMID- 9358887 TI - Estrogen-induced hypothalamic synaptic plasticity. PMID- 9358886 TI - Testing pathophysiological hypotheses of Alzheimer's disease and pharmacological modulation of amyloid precursor protein metabolism: the contribution from the studies on cultured fibroblasts from affected donors. PMID- 9358888 TI - Age-related changes in gonadotropin secretion. PMID- 9358889 TI - Melatonin in aged women. Possible modulation by estrogens. PMID- 9358891 TI - Post-menopausal changes and dementing illness: ovarian steroids as the causal link? PMID- 9358890 TI - Aging female brain: is there a change of adaptive stress? PMID- 9358892 TI - Melanocortins, neural plasticity and aging. PMID- 9358894 TI - DHEA replacement therapy for human aging: a call for perspective. PMID- 9358893 TI - Pituitary-adrenal responses to dexamethasone and CRH administration in patients with untreated Parkinson's disease. PMID- 9358895 TI - Growth hormone treatment in aging: state of the art and perspectives. PMID- 9358897 TI - Senile osteoporosis: pathophysiology and therapeutic perspectives. PMID- 9358899 TI - Electroacupuncture and laser stimulation treatment: evaluated by somatosensory evoked potential in conscious rabbits. AB - Tooth pulp generated somatosensory evoked potential (TPSEP) recordings were used to evaluate analgesic responses to Electroacupuncture (EA) and laser stimulation (Ls) treatments in Dutch-hybrid male rabbits. TPSEP recorded 100 trials at 30 minute intervals for 120 minutes. Three groups, EA, Ls and control were analyzed. The EA group received intermittent 4 Hz, intensity 2-10 volts electro-stimulation for twenty minutes. In the Ls group, a gallium aluminium arsenide (GaAlAs) laser diode with wavelength 780 nm, switched pulse frequency 9720 Hz, energy density 0.6 J/point was employed. Both EA and Ls groups received stimulation of the points Ho-ku (LI-4) and Tzu-San-Li (ST-36). Results showed tooth pulp generated noxious stimulation produced a consistent late near-field SEP waveform, which is similar to those recorded in humans and correlated to the sensation of pains, thus confirming that tooth pulp stimulation is a reliable dolormetric index. EA and Ls TPSEP recordings showed decreased peak-wave amplitude in late near-field components. These decreases correlate to analgesia. This technique provides an objective and reliable dolormetric index. PMID- 9358896 TI - Neurotrophic factors, neuroprotection and hypothalamic function. PMID- 9358900 TI - Effects of electroacupuncture and transcutaneous electrical nerve stimulation on survival of musculocutaneous flap in rats. AB - The effects of electroacupuncture (EA) and transcutaneous electrical nerve stimulation (TENS) were investigated in the musculocutaneous flap in a rat model by measuring the surviving area and blood flow in the flap. Rats were divided into the control group, and groups treated with EA and TENS. Experimental results of this study show that flap survival area did not increase by EA but increased significantly by TENS treatment, and that blood flow in the periphery was significantly larger than that at the base. PMID- 9358898 TI - Experimental evidence of a plant meridian system: V. Acupuncture effect on circumnutation movements of shoots of Phaselus vulgaris L. pole bean. AB - When the first unifoliolate leaf of Phaselus vulgaris L. cv. Kentucky wonder pole bean was at the expended stage, two needles were inserted into opposite sides of the stem near the unifoliolate bud and left there for the entire experiment. Changes of the period of circumnutation movement at the shoots of Phaselus vulgaris L. pole bean were measured. Results from two separate experiments are reported in this communication. In the first experiment, the mean period of the ultradian rhythms of the horizontal circumnutation movement of shoots was reduced significantly (p = 0.0022) from 124.2 minutes in controls to 116.3 minutes in the treated plants. The average period of the ultradian rhythms of circumnutation movement in the treated and the untreated groups from the second experiments were 96.7 and 132.1, minutes respectively, which was statistically significant (p < 0.0001). This study demonstrates for the first time that acupuncture markedly shortens the period lengths of the ultradian rhythms of circumnutation movement of the shoots of the Phaselus vulgaris L. pole bean. PMID- 9358901 TI - Thermal and antiradical properties of indirect moxibustion. AB - The thermal and antiradical properties of indirect moxibustion stimulation were investigated by thermal qualitative and spectroscopic methods. The thermal effect of indirect moxibustion was mainly dependent on the spacing distance between the moxa and skin, and not on the moxa weight. The radical scavenging activities of moxa and moxa-tar were measured by a photometric absorbance method, chemical reaction with 1,1-diphenyl-2-picrylhydrazyl. The obtained results indicate that the inhibitory effects of moxa and moxa-tar on superoxide production are due to the radical scavenging mechanism. PMID- 9358902 TI - Changes in alpha wave and state anxiety during ChunDoSunBup Qi-training in trainees with open eyes. AB - We investigated the effects of ChunDoSunBup (CDSB) Qi-training, one of the Korean popular Qi-training systems, on EEG patterns, activation coefficients and state anxiety in 13 trainees with open eyes. CDSB Qi-training procedure consists of 3 stages: sound exercise, reciting Chunmoon, which is similar to a mantra; haeng gong, a kind of body motion; and mediation. Compared to the control state (resting state before Qi-training), subjects reported less state anxiety, their activation coefficients decreased significantly during sound exercise and meditation in the occipital regions. Mean relative power and changes of mean absolute power of alpha wave increased significantly during sound exercise and meditation in the occipital regions. These results suggest that sound exercise and meditation in ChunDoSunBup Qi-training may reduce activation of the visual cortex and influence the thalamus and other functions of the brain. These could reduce anxiety levels and modulate the psychological, neurological, and physiological functions in man. PMID- 9358903 TI - A substance isolated from Cornus officinalis enhances the motility of human sperm. AB - The effects of a Chinese herb, Cornus officinalis, on the motility of human sperm was studied. An aqueous extract was prepared from the dried fruits of the herb and used in this study. The crude extract at a final concentration of 0.5 microgram/microliter in phosphate buffered saline (pH 7.4) increased sperm motility from 25.8 +/- 7.7% to 42.8 +/- 10.3% (i.e. 68% increase, n = 7), as determined by the computer-aided-sperm-analysis (CASA) method. The crude extract was fractionated by high-performance liquid chromatography (HPLC) into four fractions: C1, C2, C3 and C4. Their effects on sperm motility were further studied by CASA. Only the C4 fraction showed substantial stimulatory effects on sperm motility. At a concentration of 5 ng/microliter, C4 increased the sperm motility from 15.7 +/- 3.8% to 34.5 +/- 6.4% (i.e. 120% increase, n = 6) by CASA and from 14.9 +/- 4.3 to 28.5 +/- 8.1 (i.e. 91% increase, n = 8) by transmembrane migration ratio (TMMR) method. This result suggests that C4 is the active component in Cornus officinalis that enhances sperm motility. PMID- 9358904 TI - Superoxide anion scavenge effect of Quercus glauca Thunb. in whole blood of patients with ankylosing spondylitis. AB - Nine phenolic compounds, catechin (1), epicatechin (2), gallocatechin (3), epigallocatechin (4), procyanidin B-4 (5), catechin-3-O-rhamnoside (6), rutin (7), querglanin (8) and isoquerglanin (9) were isolated from oak leaves (Quercus glauca Thunb. Fagaceae), and the latter two (8, 9) were identified as new compounds. Several Quercus species have been used in folk medicine as an astringent for hemorrhoids and for treatment of inflammation, jaundice, and tumor. In this study, these compounds were tested for scavenging effects of the superoxide anion in the whole blood of patients with ankylosing spondylitis by means of an ultra-sensitive chemoluminescence (CL) analyzer and lucigenin amplification. The results showed that at a concentration of 2.3 x 10(-5) M, isoquerglanin (9) displayed the strongest inhibition activity (73.55%), followed by querglanin (8) (68.81%) and then gallocatechin (3) and epigallocatechin (4) (66.97 and 60.17% inhibition, respectively). In addition, the blood chemoluminescence (CL) level of patients with ankylosing spondylitis was inhibited by superoxide dismutase (SOD) but not by catalase, suggesting that superoxide anion is the major component of reactive oxygen species (ROS) involved in this assay system. PMID- 9358905 TI - Effect of gegen-tang on painful gynecomastia in patients with liver cirrhosis: a brief report. AB - Four patients with liver cirrhosis and complaining of painful gynecomastia were treated with oral administration of Gegen-Tang (TJ-1). Pain disappeared in 3 patients in one week, and in one patient in 4 weeks. The size of gynecomastia did not change significantly on mammography, but palpable induration diminished or disappeared. The patients had been treated with Chaihu (saiko) group drugs for liver diseases, and TJ-1 was used in combination with these drugs. Serum levels of estrogen, progesterone, testosterone, and other sex hormones did not change significantly after TJ-1 treatment. These results suggest that TJ-1 could be used for the painful gynecomastia that is occasionally seen in cirrhotic patients. PMID- 9358906 TI - The hepatoprotective and therapeutic effects of propolis ethanol extract on chronic alcohol-induced liver injuries. AB - Propolis designates a mixture of gums, resins and balms, of viscous consistency, which are gathered on certain parts (buds and bark, mainly) of vegetables (especially coniferous trees) by honeybees. They bring this back to the hive, where it is modified and mixed with other substances (essentially their own wax and salivary secretions). In this study, the hepatoprotective and therapeutic effects of propolis ethanol extract on chronic alcohol-induced liver injuries were investigated in rats. 3.125 ml of 99.5% alcohol was added to animal's daily diet for four weeks to induce chronic alcohol liver injuries. After sacrifice, serum transaminases (GOT, GPT), triacylglyceride and hepatic triacylglyceride (HTG) concentration were assayed to observe liver injuries induced by chronic alcohol abuse. In addition, the phenomenon of alcohol induced fatty liver were also observed by histopathological changes. Different doses of propolis ethanol extract were p.o. administered three times per day for three days, after four weeks' alcohol administration. It was found that 10 mg/kg of propolis ethanol extract significantly decreased the elevations of serum GOT, GPT, TG and HTG. In histopathological examination, 30 mg/kg of propolis ethanol extract also remarkably decreased the hepatocellular fatty degeneration, apparent as vacuolization, induced by chronic alcohol abuse. PMID- 9358907 TI - Comparative study of oral and parenteral administration of sho-saiko-to (xiao chaihu-tang) extract on D-galactosamine-induced liver injury in rats. AB - The preventive effect of Sho-saiko-to (Xiao-Chaihu-Tang) extract (TJ-9) on the progression of D-galactosamine (GaIN)-induced liver injury was examined in five week-old male Wistar rats with oral (p.o.) or intraperitoneal (i.p.) administration of the same dose of TJ-9. Rats treated once with GaIN (500 mg/kg body weight, i.p.) received TJ-9 at a dose of 1.0 g/kg body weight (p.o. or i.p.) 2 hours after GaIN treatment at which time an apparent liver injury occurred. Both p.o. and i.p. administration of TJ-9 showed similar significant prevention against the progression of liver injury 24 hours after GaIN injection. Although total protein and albumin concentrations in serum and protein concentration in the liver decreased with the progression of GaIN-induced liver injury, oral or i.p. administration of TJ-9 prevented these decreases in similar degree. However, decreases in serum and liver triglyceride concentration with the progression of liver injury were not attenuated after p.o. or i.p. administration of TJ-9. The activities of liver 5'-nucleotidase and glucose-6-phosphatase, marker enzymes of liver plasma and microsomal membranes, respectively, decreased during the progression of liver injury. A similar preventive effect on the decrease of both enzyme activities was found after p.o. or i.p. administration of TJ-9. These results indicate that the preventive effect on progression of GaIN-induced liver injury by oral or i.p. administration is approximately equal, and that the effect may be through improving the impaired liver protein synthesis and disrupted liver plasma and microsomal membranes in a similar degree. PMID- 9358908 TI - Biphasic effects of pu-chung-i-chi-tang on sedation and excitation. AB - The biphasic effects of Pu-Chung-I-Chi-Tang (PCT) on sedation and excitation in acute treatment or after one-week consecutive treatment were studied. The results indicated that PCT produces sedation in acute treatment and excitation after one week consecutive treatment. The sedative mechanism of PCT in acute treatment might be due to an increase in serotonergic activity and a decrease in dopaminergic activity. However, the excitatory mechanism of PCT after one-week consecutive treatment might involve the increase in dopaminergic activity and the decrease in serotonergic activity. PMID- 9358910 TI - Pulse spectrum study on the effect of sie-zie-tang and Radix aconiti. AB - Extracts of the traditional Chinese formula Sie-Zie-Tang as well as one of its main components, Radix Aconiti were injected into rats intraperitoneally to observe pressure wave spectrum changes at the caudate artery. We found that Radix Aconiti decreased the C0 (DC term of the pulse), C5 and C6 (the harmonic proportions of the 5th and the 6th harmonic), but increased C2 and C3 (the harmonic proportions of the second and the third harmonic) significantly. For Sie Zie-Tang, the increases of C2, C3, and C4 were accompanied by the decreasing of C0. The decreases of C5, C6 were small and not significant. The additional ingredients in the formula reduce toxic side effects (arrhythmia or heart failure caused by faster and stronger heart beat) due to Radix Aconiti. For human subjects, low dose Sie-Zie-Tang tends to normalize the Fourier components of the pressure wave. Orally taking the formula elevates the harmonic proportion of the harmonic that is lower than normal, but suppresses the higher one. Our results provides a possible mechanism for heart meridian related herbs. It strengthens heart beats, and normalizes energy distribution to different meridians. The study on Sie-Zie-Tang reveals another formula construction to reduce toxic side effects. PMID- 9358909 TI - The use of Chinese herbal medicine on experimental fracture healing. AB - The purpose of this study was to examine the effect of Ru-Yih-Jin-Huang-Saan (RYJHS) and Jie-Guu-Saan (JGS) on experimental fracture healing in rats. Seventy five rats were fractured in the middle of the left tibia and fibula. They were randomly divided into experimental and control groups and injected with Ringer's solution and applied with Ru-Yih-jin-Huang-Saan externally or fed with Jie-Guu Saan respectively. The animals were sacrificed at 4, 8 and 12 days after fracture. The contents of hydroxyproline and the synthesis of DNA, RNA and protein were observed. The results demonstrate that groups treated with herbal medicine were more rapidly and thoroughly healed than the control group in respect to collagen formation and bone cell metabolism. PMID- 9358911 TI - Factors associated with the utilization of traditional Chinese medicine in a small town in Hong Kong. AB - The practice of Traditional Chinese Medicine (TCM) is largely unregulated in Hong Kong. Yet, as previous studies have shown, a sizable segment of the population consults TCM practitioners for health problems. This paper uses health care utilization data from a telephone health survey of 847 adult subjects in Tai Po District who had suffered from acute illness in the past month, to examine the profile of TCM users in the District. Women, older residents, unemployed workers, low skill laborers, current smokers and subjects dissatisfied with the quality of private sector clinics were significantly more likely to consult TCM practitioners. PMID- 9358912 TI - The CARET asbestos-exposed cohort: baseline characteristics and comparison to other asbestos-exposed cohorts. AB - The Carotene and Retinol Efficacy Trial (CARET) was a double-blind, placebo controlled trial of the daily administration of 25,000 IU vitamin A and 30 mg beta-carotene for the prevention of lung cancer. Of close to 18,500 participants, more than 4,000 were asbestos-exposed men recruited from shipyard and construction trades at five study centers in the United States. While the primary endpoint of the trial was the incidence of lung cancer, a number of questions about the natural history of asbestos-related disease will also be addressed. The mean age at entry into the trial was 57 years and the mean duration of follow-up on active intervention was 4 years. With the exception of 133 never-smoker pilot participants (3%), all subjects recruited were by intention current (38%) or ex smokers (58%), with a mean cumulative smoking exposure at entry of 43 pack-years. Mean years from first asbestos exposure were 35, and mean duration of asbestos exposure in a high-risk trade was 19 years. The distribution of radiographic abnormalities was as follows: normal, 34%; parenchymal opacities (ILO profusion score > 1/0) alone, 18%; pleural thickening alone, 27%; both parenchymal opacities and pleural thickening, 21%. The CARET cohort, when compared to previously reported asbestos-exposed cohorts, is characterized by substantial asbestos exposure and high proportion of asbestos-related radiographic findings. The active intervention was halted in 1996, after a mean duration of 40 years. Passive follow-up of the cohort will continue until the year 2000. PMID- 9358914 TI - Major and trace element compositions of the UICC standard asbestos samples. AB - Major and trace element compositions for chrysotile (2 samples), amosite, crocidolite, and anthophyllite UICC standard asbestos samples have been determined using UV-visible spectrophotometry, atomic absorption spectometry, flame photometry, volumetric analysis, and gravimetric analysis for major elements and x-ray and optical spectrometry for trace elements. The trace element data are for Li, S, Cl, Sc, V, Cr, Co, Ni, Cu, Zn, Ga, Rb, Sr, Zr, Nb, Ba, La, Ce, Pb, and Th and distribution in the various mineral phases is discussed. PMID- 9358913 TI - Lobe of origin and histologic type of lung cancer associated with asbestos exposure in the Carotene and Retinol Efficacy Trial (CARET). AB - Lower lobe origin and histologic diagnosis of adenocarcinoma have been described as useful parameters for attributing lung cancer to prior asbestos exposure. To assess whether these characteristics differed between asbestos-exposed individuals and smokers, we evaluated lobe of origin and histologic type of tumors in 78 asbestos-exposed and 214 nonexposed heavy smokers developing lung cancer during the Carotene and Retinol Efficacy Trial (CARET), a prospective cancer chemoprevention trial. Most tumors in both cohorts, regardless of radiographic fibrosis at baseline, originated in upper lobes, representing 67% in asbestos-exposed and 80% in smokers, respectively (adjusted OR for lower lobe = 1.41; 95% CI = 0.69-2.91). Adenocarcinoma represented 32% of lung tumors in the asbestos cohort, and 30% in the smoking cohort (adjusted OR = 0.78; 95% CI = 0.40 1.55), and was inversely associated with radiographic fibrosis (adjusted OR = 0.19; 95% CI = 0.06-0.62). We conclude that neither anatomic site nor histologic cell type of tumors distinguishes effectively between smoking and asbestos as causal factors in development of lung cancer. PMID- 9358915 TI - Silicosis, mixed dust pneumoconiosis, and lung cancer. AB - A total of 764 autopsy cases with a pathological diagnosis of nonasbestos pneumoconiosis were investigated in a search for lung cancer: 146 patients bore 148 lung cancers (19.1%). The incidence of a lung cancer was associated positively with aging longer occupational exposures, and smoking habits. A reverse correlation was found between carcinogenesis and the severity of pneumoconiosis. A statistically significant increase in the incidence of certain types of lung cancer (squamous cell carcinoma + small cell carcinoma) was found in silicotic lungs with massive fibrosis as compared to lungs with mixed dust pneumoconiosis of comparable severity. Although there appears to be no dose response relationship in general between silicosis and lung cancer, it is advisable to consider the possibility that a presumptive silica-induced carcinogenesis might be masked by the severe fibrosis of a silicotic type, which obliterates the lung tissue in a different way from asbestosis, which is associated with epithelial proliferation. PMID- 9358916 TI - Tuberculin reactivity among California Hispanic migrant farm workers. AB - We conducted a cross-sectional study of tuberculin reactivity among residents of two northern California migrant-farm-worker housing centers. Participants completed a brief health questionnaire and were offered tuberculin skin testing with radiologic and medical follow-up. Four hundred and sixty-nine persons (estimated participation rate: 70%) completed questionnaires. All but one were Hispanic. Two hundred and ninety-six (63%) participants completed tuberculin skin testing and 49 (16.6%) showed reactivity (> or = 10 mm induration at 48-72 hours). Increased prevalence was seen for the 15-39-year age group (vs. persons younger than 15: OR 2.59; 95% CI 0.79-8.47), former smokers (vs. never smokers: OR 3.11; 95% CI 1.20-8.09), and persons born outside the U.S. (OR 2.09; 95% CI 0.66-6.61). Prophylaxis with isoniazid was recommended for 23 persons; nine (39%) completed therapy. No cases of active tuberculosis were found. Prevalence of tuberculin reactivity in this population is lower than reported among Hispanic farm workers in the eastern and midwestern U.S. Higher prevalence may obtain among California farm workers not included in the study population, including homeless, single, and highly mobile persons. Public-health efforts in this population should focus on ever-smokers, young adults, and persons born outside the U.S. PMID- 9358917 TI - Correlations among human blood levels of specific PCB congeners and implications for epidemiologic studies. AB - Specific congeners of PCBs may differ with respect to their human health risks. For epidemiologic studies, however, measuring levels of specific congeners--as compared with estimating the concentration of total PCBs present, may be of limited value if levels of specific congeners are highly correlated. We examined the correlations among levels of specific congeners in three groups: controls from a case-control study of breast cancer in North Carolina and two groups from Wisconsin with exposure to fish from contaminated waters. Levels of specific congeners were, in general, highly correlated (Pearson r > 0.80). However, the level of congener 180, a heptachlorobiphenyl, tended to be less correlated with levels of lower-chlorinated biphenyls. Among the implications of these findings are that measurement of a select group of congeners may yield essentially the same information as measurement of a large panel, and may be more cost efficient. PMID- 9358918 TI - Cancer mortality among laundry and dry cleaning workers. AB - A cancer mortality study of 8,163 deaths occurring among persons formerly employed as laundering and dry cleaning workers in 28 states is described. Age adjusted sex-race cause-specific proportionate mortality ratios (PMRs) and proportionate cancer mortality ratios (PCMRs) were computed for 1979 through 1990, using the corresponding 28-state mortality as the comparison. For those aged 15-64 years, there were excesses in black men for total cancer mortality (PMR = 130, 95% confidence interval (CI) = 105-159) and cancer of the esophagus 1 (PMR = 215, 95% CI = 111-376), and in white men for cancer of the larynx (PMR = 318, 95% CI = 117-693). For those aged 65 years and over, there were statistically nonsignificant excesses for cancer of the trachea, bronchus, and lung in black women (PMR = 128, CI = 94-170) and for cancer of other and unspecified female genital organs in white women (PMR = 225, CI = 97-443). The results of this and other studies point to the need for the effective implementation of available control measures to protect laundry and dry cleaning workers. PMID- 9358919 TI - Upper limb work-related musculoskeletal disorders among newspaper employees: cross-sectional survey results. AB - At a metropolitan newspaper office in Canada with extensive video display terminal (VDT) use, researchers carried out a survey (n = 1,007, 84% response) to establish baseline prevalence of work-related musculoskeletal disorders (WMSDs) and to identify demographic, postural, task, and psychosocial factors associated with WMSD symptoms. One-fifth of the respondents reported moderate or worse upper limb pain recurring at least monthly or lasting more than a week over the previous year. Logistic regression showed that employees who faced frequent deadlines and high psychological demands (fast work pace and conflicting demands), had low skill discretion and social support, spent more time keyboarding, or who had their screen in a non-optimal position were more likely to report moderate to severe symptoms. Women reported significantly higher levels of symptoms than men. PMID- 9358920 TI - Surveillance of work-related musculoskeletal injuries among union carpenters. AB - Combined data sources, including union administrative records and workers' compensation claims, were used to construct event histories for a dynamic cohort of union carpenters from Washington State during the period 1989-1992. Person time at risk and the events of interest were stratified by age, sex, time in the union, and predominant type of carpentry work. Poisson regression techniques were used to identify subgroups at greatest risk of filing claims for a variety of musculoskeletal disorders defined by ANSI codes for body part injured and injury nature. Distinguishing different kinds of musculoskeletal disorders, even crudely with ANSI codes, led to different conclusions about the effects of the explanatory variables. Among older workers, the rates of fractures of the foot were higher, while rates of contusions of the hand and foot were lower. Women had higher rates of sprain/strains and nerve conditions of the wrist/forearm. Higher rates of injuries to the axial skeleton were seen among carpenters who did predominantly light commercial and drywall work, while piledrivers had lower rates of these injuries. Drywall workers had higher rates of sprains to the ankle/lower leg. Workers who were members of the union as long as four years had lower risks for the vast majority of musculoskeletal disorders studied. Similar patterns were seen for more serious claims that resulted in paid lost time from work. PMID- 9358921 TI - An ergonomic education and evaluation program for apprentice carpenters. AB - Eighteen new apprentice carpenters received sixteen hours of ergonomics awareness education as a part of their regular apprenticeship training during 1994 and 1995. An equal number of apprentices received no training but served as controls. The training took place in the Southwest Ohio District Council of Carpenter's Joint Apprenticeship and Training School. The curriculum was designed to be "learner-centered." Instruction included short lectures presented by a journeyman carpenter and emphasized participatory activities in the school's carpentry shop. Ongoing program evaluation assessed trainees' reactions to the content and structure of the curriculum and its influence on their behavior. Trainees and controls completed brief quizzes on ergonomic knowledge. Hands-on exercises enabled trainees to apply recently acquired ergonomic knowledge in the school's carpentry shop. Trainees scored significantly higher on one-half of the post session quizzes and the comprehensive test. Trainees preferred participatory teaching methods, especially those using redesigned tools (93%) and evaluating ergonomic risks (86%); and they supported continued safety and health education during apprentice training. The authors conclude that apprentice-ship programs should provide regular "learner-centered" occupational safety and health education that includes ergonomics, and these programs should be integrated with their shop-based manual arts instruction. PMID- 9358922 TI - Injury severity associated with nonfatal construction falls. AB - This study evaluated injury severity in a group of construction workers who sustained nonfatal falls at work. The sample consisted of 255 adults who were identified from Doctor's First Reports (DFRs) submitted to the California Department of Industrial Relations. For those that fell from heights (n = 195), the mean height of fall was 9.2 feet (SD = 7.1). The mean number of lost work days was 44.3 days (SD = 58.6) and the median was 10 days. Two measures of injury severity were used--the Injury Severity Score and the disability section of the Health Assessment Questionnaire (HAQ). Seventeen participants (7%; 95% CI, 4-10%) were deemed permanently disabled. A simultaneous multiple regression model, using five independent variables, explained approximately 21% of the variance in HAQ scores. Nonunion status and safety climate scores indicating increased risk were positively correlated with higher functional limitation as measured by HAQ scores, as were greater heights and impact on concrete surface. Higher scores on both injury severity measures were significantly and moderately associated with a greater number of days lost from work. These findings suggest that injury severity and permanent disability associated with falls in construction are notable, and identify key target areas for intervention and prevention. PMID- 9358923 TI - Nocturnal oxygen desaturation, as assessed by home oximetry, in long-term solvent exposed workers. AB - Recent studies have suggested that occupational exposure to solvents may be a cause of sleep apnea. Digital oximetry during one night was performed in solvent exposed offset printers (n = 21) and in a control group (n = 21), using a Palco 400 Pulse Oximeter. The threshold for recording was set at an arterial oxygen saturation (SaO2) of 90%. Furthermore, computerized neurobehavioral tests (NES) and a solvent-related complaints questionnaire (NSC-60) were administered. The mean exposure time was 15 years (SD = 10). Hygiene measurements revealed a large number of different solvents and a cumulative exposure between 15% and 97% of the "cumulative TLV." The exposed workers had more solvent-related complaints, especially regarding mood (analysis of covariance, P = 0.02), than the nonexposed workers. The neurobehavioral tests indicated that hand-eye coordination was significantly worse in the exposed group (analysis of covariance, P = 0.03). The frequency of nocturnal desaturation was significantly higher in the printers (1.7 events/hr +/- SD = 1.5) than in the controls (0.6 events/hr +/- SD = 1.3) (Mann Whitney test, P < 0.01). Also, the duration of desaturation was longer in the exposed workers: 3.2 min/hr (SD = 3.2) vs 1.2 min/hr (SD = 2.3) (Mann-Whitney test, P < 0.01). In the analysis of covariance, exposure (P = 0.04) and the interaction between smoking and exposure (P = 0.02) were shown to contribute significantly to the excess of nocturnal desaturation in the exposed. The same was true for the mean duration of desaturation (exposure: P = 0.02 and interaction exposure smoking: P = 0.02). The significant interaction was due to a more pronounced effect of solvent exposure among the nonsmoker group. No relation was found between the excess of complaints or the neuroperformance effects and the oximetry data. These data reinforce the presumption that occupational solvent exposure might contribute to sleep-disordered breathing. PMID- 9358924 TI - Prospective epidemiologic evaluation of laboratory animal allergy among university employees. AB - OBJECTIVES: Evaluation of incidence and risk factors for development of laboratory animal allergy (LAA) among new hires previously unexposed to lab animals. METHODS: Baseline, 6-month and yearly follow-up, questionnaires, pulmonary functions, and methacholine challenges were collected from 98 never before occupationally exposed and 90 control laboratory researchers. The two groups were followed between 6 and 36 months. RESULTS: At baseline, there were no differences in atopy, pulmonary functions, or methacholine reactivity between the two groups. The incidence of work-related asthma was comparable in the two groups, approximately 2.5% at 6 months and 4.5% at 24 months. The rate of decline in FEV1 was statistically significantly greater in the animal-exposed than nonanimal-exposed workers, and animal-exposed smokers' FEV1 declined significantly more rapidly than any other groups'. CONCLUSION: Despite the low incidence of laboratory-animal allergy and work-related asthma in this group, this study corroborates previously described interaction between smoking and animal exposure. PMID- 9358925 TI - Blood interleukin-8 production is increased in chemical workers with bronchitic symptoms. AB - Chemical exposure may result in respiratory conditions such as chronic bronchitis, bronchial hyperresponsiveness, and chronic airway obstruction. Clinical studies have shown that during the course of disease, cytokine networks are changed. In order to study the relationship between blood cytokines and respiratory symptoms in an occupational setting, we investigated 106 chemical workers during a routine yearly medical examination in 1995. Lung function was measured with flow volume curves and impedance using the forced oscillation technique (FOT). Smoking-status and respiratory symptoms were determined by questionnaires. Cytokines were selected on biological plausibility and measured both in a whole blood assay (TNF-alpha, IL-8) and in serum (IL-4, IL-5, IL-6, IFN gamma). The hypothesis is that blood levels of TNF-alpha and IL-8 are increased in bronchitis, while serum levels of IL4, IL-5 are increased and IFN-gamma is decreased in asthmatic workers. Spontaneous IL-8 release was significantly higher in workers with bronchitis (P < 0.05) or chronic bronchitis (P < 0.01) compared to workers without those respiratory symptoms, also after correction for age, pack-years, and blood lymphocyte numbers or compared to a matched control group. No correlation was present between specific cytokines and asthmatic symptoms. These data suggest that blood IL-8 may be considered as a useful marker for bronchitis. PMID- 9358926 TI - Immunological effects of CaEDTA injection: observations in two lead workers. AB - To evaluate the effects of calcium disodium ethylenediamine tetraacetate (CaEDTA) injection on human immune system in relation to exposure to lead, we administered CaEDTA by intravenous injection for 1 hr three times (three consecutive days) a week to two male lead workers. They had been engaged in recycling lead for 31 and 22 years, aged 61 and 53 years (workers 1 and 2), respectively. Before the treatment of CaEDTA, their blood lead concentrations (PbB) were 81 and 68 micrograms/dl, respectively. The administration of CaEDTA had been carried out to worker 1 for 10 weeks and to worker 2 for 6 weeks. A significant decrease in PbB between before and after three-times CaEDTA injection was found in both workers. Significant increases in IgG, IgA, IgM, CD8+, and CD57+ cells were found in worker 1. A significant increase in IgD was found in worker 2. During the study period, IgG in worker 1 and CD4+ cells in worker 2 were gradually increasing. There was a significant negative correlation between IgG and PbB in worker 1. It is suggested that the immunological function such as antibody formation in lead workers might be improved by CaEDTA injection. PMID- 9358927 TI - Work with video display terminals and the risk of reduced birthweight and preterm birth. AB - To determine whether the use of video display terminals (VDTs) is associated with an increased risk of reduced birthweight (RBW) and preterm birth, a cohort of telephone operators who used VDTs at work was compared to a cohort of non-VDT users. Among 2,430 women interviewed, 713 eligible singleton live births were reported. Exposure was estimated from company records and a representative sample of electromagnetic fields was measured at the VDT workstations. For RBW (< or = 2,800 g), we found no excess risk associated with any VDT use during pregnancy (odds ratio [OR] = 0.9; 95% confidence interval [CI] = 0.5-1.7). For preterm birth (< or = 37 weeks), we similarly found no excess risk (OR = 0.7; 95% CI = 0.4-1.1). The risks estimated did not change substantially when hours working with VDTs were used as exposure variables. By contrast, increased risks were found for several known risk factors for LBW and preterm birth. We conclude that occupational VDT use does not increase the risk of RBW and preterm birth. PMID- 9358928 TI - TV broadcast towers and cancer: the end of innocence for radiofrequency exposures. PMID- 9358929 TI - Comparison of single photon emission computed tomography findings in cases of healthy adults and solvent-exposed adults: correction of previous results. PMID- 9358930 TI - Re: Comparison of single photon emission computed tomography findings in cases of healthy adults and solvent-exposed adults. PMID- 9358931 TI - How threshold limits for lead were established in the 1950s. PMID- 9358932 TI - The other information revolution. PMID- 9358933 TI - Quality of life assessment in patients receiving adjuvant therapy for breast cancer: the IBCSG approach. The International Breast Cancer Study Group. AB - BACKGROUND AND PURPOSE: The International Breast Cancer Study Group (IBCSG) has developed and approach for assessing the impact of adjuvant therapy on quality of life (QL) within the framework of international, multilingual clinical trials. The major steps are summarized. Conceptual, methodological and practical issues are discussed with reference to results of two trials closed to accrual (IBCSG VI, VII) and one subsequent ongoing trial (IBCSG IX). PATIENTS AND METHODS: QL was assessed in pre- and post-menopausal patients with operable breast cancer. Various single-item linear analogue self-assessment (LASA) scales were used as indicators of components of QL, including global indicators of well-being, functioning and health perception, and specific indicators of symptoms of disease and treatment. In trials VI and VII, QL was assessed at baseline, during adjuvant treatment and follow-up, and at recurrence. Based on this experience, the QL form was revised for subsequent trials and further investigated in a subsample of patients randomized into trial IX. RESULTS: In trials VI and VII, the QL indicators were responsive to the impact of biomedical factors at baseline, various adjuvant treatments, changes over the first 18 months, and recurrence. In trial IX, the revised QL form was well accepted by patients and staff. Completing this form did not exceed five minutes. QL differences between on and off cytotoxic treatment strengthen the claim that these measures are responsive. Correlations and logistic regression analyses show the expected relationship among the various global and specific indicators. CONCLUSION: Results from two trials closed to accrual and an ongoing trial confirm the feasibility, validity and clinical relevance of the IBCSG approach for studying the impact of adjuvant breast cancer therapy on QL in international clinical trials. PMID- 9358934 TI - Topoisomerase I inhibitors: review and update. AB - This review presents a summary of preclinical and clinical data on the topoisomerase I (topo I) inhibitors that are under clinical development. To date, all of the topo I inhibitors that have been clinically evaluated are analogues of camptothecin, an extract of the Chinese tree Camptotheca acuminata. The therapeutic development of camptothecin was initially limited by its poor solubility and unpredictable toxicity. More recently, a number of water-soluble camptothecin analogues have undergone extensive evaluation and have demonstrated significant clinical activity. These include irinotecan (CPT-II), topotecan, and 9-aminocamptothecin (9-AC). Preliminary data are also reviewed on other camptothecin analogues (GG-211 and DX-8951f), on oral formulations, and on non camptothecin topoisomerase I inhibitors. The topoisomerase I inhibitors have already demonstrated a broad spectrum of antitumour activity, most probably due to their unique mechanism of action and lack of clinical cross-resistance with existing antineoplastic compounds. The challenge for the next five years is to identify ways to integrate the topo I inhibitors into multidrug and multimodality therapies to achieve optimal antitumour effect, while keeping the side effects of these therapies manageable. PMID- 9358935 TI - The dynamics of change: cancer patients' preferences for information, involvement and support. AB - BACKGROUND: While the importance of providing individualised communication to cancer patients is now well recognised, little is known about the stability and validity of patients' expressed preferences for information and involvement in decision-making. This study explored the stability and possible predictors of such preferences over time. PATIENTS AND METHODS: Cancer patients seeing two Medical Oncologists in an out-patient clinic at an Australian teaching hospital completed a questionnaire battery before and directly after one consultation, and before their next consultation. Eighty consecutive patients with heterogeneous cancers participated in the study. Preferences for general and specific information, involvement and support were elicited at each assessment. Locus of control and patient familiarity with the clinic were measured before the first consultation. Patient satisfaction with the consultation was assessed directly after the consultation. Demographic and disease data were recorded for each patient. RESULTS: General preferences for information and involvement were relatively stable, at least in the short term; however there was considerable variability in preferences for specific topics of information. Patients whose condition had recently worsened were more likely to want progressively less involvement in decision-making. Gender, the doctor seen and religion were also predictive of patient preferences. CONCLUSIONS: Situational factors, such as change in disease status, may alter a patient's preferences for information and involvement. If we wish to match the provision of information and support to the expressed needs of patients, we must ask patients at each consultation what those needs are. PMID- 9358936 TI - Phase I-II study of vinorelbine in combination with 5-fluorouracil and folinic acid as first-line chemotherapy in metastatic breast cancer: a regimen with a low subjective toxic burden. AB - BACKGROUND: Vinorelbine, is an active drug in the treatment of metastatic breast cancer and has a favorable toxicity profile. Its combination with other effective and well-tolerated cytotoxics may thus be beneficial. We investigated the therapeutic effect of a combination of vinorelbine plus 5-fluorouracil and folinic acid as first-line treatment in patients with metastatic breast cancer. PATIENTS AND METHODS: Forty-five patients with advanced or metastatic breast cancer were enrolled in this phase I-II study and treated with 5-fluorouracil (350 mg/m2 i.v. on day 1 to 3), folinic acid (100 mg/m2 i.v. on day 1 to 3) and vinorelbine given on days 1 and 3 at the dose of 25 mg/m2 (dose level 1), or 30 mg/m2 (dose level 2). Therapy was given on an outpatient basis every three weeks. RESULTS: Phase I: Dose limiting toxicity (DLT) occurred at the second dose level of vinorelbine (30 mg/m2), with two out of three patients developing severe constipation ('ileus-like syndrome' grade 4), and fever (grade 2). Consequently, the dose evaluated in the phase II study was 25 mg/m2. Phase II: Objective responses were observed in 24 of 39 evaluable patients (95% confidence interval (95% CI), 47% to 77%). There were seven complete responses (18%), 17 partial responses (44%), and for nine patients (23%) disease was stable. Only six patients (15%) experienced disease progression. The median response duration was 10 months (range 6 to 24+) and the median time to progression was eight months (range 2 to 24+). Granulocytopenia was the most frequently observed side effect, with a grade 4 nadir being observed in 30 patients (77%), with four hospital admissions due to febrile neutropenia. Nausea, vomiting, and anorexia were mild to moderate and reported by less than half of the patients. Alopecia was moderate and occurred in about one-third of the patients. The other side effects were mild and easily manageable. CONCLUSIONS: This effective combination chemotherapy of vinorelbine, 5-fluorouracil and folinic acid is comparable to other first-line regimens in terms of efficacy, and is subjectively well tolerated, thus deserving a test in randomized trials in the advanced and adjuvant settings. PMID- 9358937 TI - Prognosis of Kaposi's sarcoma as an initial and later AIDS associated illness. AB - BACKGROUND: The natural history of Kaposi's sarcoma (KS) as a primary presentation of AIDS has been well defined, but little is known about the prognosis of KS following a different AIDS defining illness (ADI). PATIENTS AND METHODS: Retrospective review of 852 consecutive individuals diagnosed with AIDS at Fairfield Hospital between 1984 and 1994. Demographic data, year of diagnosis, CD4 cell counts, treatment for KS and PCP prophylaxis were included in the analysis. Survival following a diagnosis of KS was evaluated, adjusting for the effects of year of diagnosis, primary or secondary KS and degree of immunodeficiency. RESULTS: The overall cumulative incidence of KS by three years post ADI was 34%. Median survival for KS as an ADI (n = 130) was 20 months versus 9 months for KS subsequent to another ADI (n = 75, P < 0.001). Those with KS as an ADI had a higher CD4 count (median 90 vs. 11, P < 0.001), lower incidence of visceral disease (5 of 130 vs. 11 of 75, P = 0.032) and fewer associated AIDS related illnesses (1 vs. 2, P < 0.001). Poorer survival following diagnosis of KS was associated with a lower CD4 count at diagnosis of KS (P = 0.002), extensive cutaneous or visceral KS at diagnosis (P = 0.009 and P < 0.001 respectively) and with the number of associated AIDS related illnesses (P < 0.001). A multivariate analysis suggested that, after adjusting for these factors, there was no difference in survival between primary and secondary KS. CONCLUSION: We found no difference in survival between primary and secondary KS after adjusting for potential confounding factors. We cannot exclude, however, that the greater incidence of visceral disease identified in secondary KS reflects an inherently more aggressive biology. PMID- 9358938 TI - Meeting patient expectations in the cancer consultation. AB - BACKGROUND: Low scores on satisfaction measures may be anticipated when patients' expectations of the doctor are unmet during the cancer consultation. We correlated discrepancies between patient expectations of their ideal doctor and their perceptions of their actual doctor with scores on a validated satisfaction scale to determine whether patients whose expectations were unmet were less satisfied. PATIENTS AND METHODS: The expectations questionnaire used a forced choice method designed to elicit patient preferences for either emotional or informational support from the physician. One hundred and five new patients with heterogeneous cancers, of five medical oncologists at a major teaching hospital were sampled. The patients were mostly female (55%) middle aged (mean age 54.3) and newly diagnosed with cancer (56% within two months prior to consultation). RESULTS: Patients did not demonstrate a clear preference for an emotionally or informationally supportive approach. Seventy percent of patients did not want emotionally negative physicians but most (88.4%) would tolerate negative information. The mean number of exact matches between patients expectations of the ideal and their perceptions of their actual doctor was 3.7 (from a total of six). 5.9% of patients received exactly the doctor they wanted. No significant differences in satisfaction were found between patients whose expectations were met and those whose expectations were not met. CONCLUSIONS: Patient satisfaction with the consultation was independent of patient expectation for informational or emotional support. PMID- 9358939 TI - Nongastrointestinal mucosa-associated lymphoid tissue (MALT) lymphomas: clinical and therapeutic features of 24 localized patients. AB - BACKGROUND: Peripheral B-cell lymphoma of the marginal zone (MALT, low-grade), presenting as localized, extranodal disease, usually affects the elderly. The gastrointestinal tract is the most frequently involved extranodal location, representing 70% of all MALT lymphomas. Recently, numerous other extranodal sites involved by MALT lymphomas have also been described. PATIENTS AND METHODS: From January 1990 to October 1995, 24 patients with untreated nongastrointestinal low grade MALT lymphoma were submitted to treatments ranging from the local approach of radiotherapy and local alpha-interferon (alpha-IFN) administration to chemotherapy. The tumours were located in the lung (seven cases), conjunctiva (four cases), lachrymal gland and orbital soft tissue (four cases), salivary glands (three cases), skin (three cases), breast (two cases), and thyroid (one case). All patients had low-grade stage IE tumours. RESULTS: Chemotherapy was administered in 11 patients (six with lung, three with salivary gland, one with breast, and one with thyroid locations); radiation therapy was employed in seven patients (three with lachrymal gland, three with skin, and one with breast locations); local alpha-IFN administration was administered in five patients (four with conjunctival, and one with lachrymal gland sites); and surgery was employed in one patient with a lung tumour. All patients achieved complete remissions; three local recurrences and two relapses in other sites were observed. The global five-year survival rate was 100% with a relapse-free survival rate of 79%. CONCLUSIONS: These data confirm the significant efficacy of different therapeutic approaches to specific sites inbes obtaining a good remission rate for nongastrointestinal localized low-grade MALT lymphomas. PMID- 9358940 TI - The efficacy of a combination of ondansetron, methylprednisolone and metopimazine in patients previously uncontrolled with a dual antiemetic treatment in cisplatin based chemotherapy. The French Ondansetron Study Group. AB - BACKGROUND: Cisplatin is one of the most effective cytotoxic drugs used in the treatment of certain neoplasms, but is also one which most frequently induces nausea and vomiting. Combination of corticosteroids with ondansetron enables greater control of emesis than that obtained with ondansetron alone, but some patients still experience symptoms. The objective of this randomised, double blind, multicentre, parallel group study was to examine the benefit of the addition of metopimazine (MPZ), a dopamine receptor antagonist, to the combination of ondansetron + methylprednisolone (O + M) in the prevention of cisplatin-induced nausea and vomiting in patients uncontrolled [i.e., at least one emetic episode (vomiting and/or retching) or moderate or severe nausea] during their previous course of cisplatin based chemotherapy, despite antiemetic treatment with a combination of a 5-hydroxytryptamine3 receptor antagonist (5HT3) with a corticosteroid. The impact of the treatment on the patients' quality of life was also evaluated using two specific questionnaires the FLIC (Functional Living Index for Cancer), and the FLIE (Functional Living Index for Emesis). PATIENTS AND METHODS: The intent-to-treat population consisted of 338 patients; 168 patients received the triple combination of ondansetron, methylprednisolone and metopimazine (O + M + MPZ), and 170 patients received ondansetron plus methylprednisolone (O + M). Tumour type was comparable in the two treatment groups, the most prevalent being lung cancer. Patients in group O + M + MPZ received ondansetron as an 8 mg intravenous injection prior to chemotherapy on day 1 followed by 8 mg tablets b.i.d. from D2 to D3, methylprednisolone as a 120 mg intravenous injection prior to chemotherapy on D1 and followed by 16 mg tablets b.i.d. from D2 to D3, and metopimazine as a 40 mg intravenous injection prior to chemotherapy on D1 and followed by 15 mg capsules b.i.d. on D2 to D3. Patients in group O + M received treatment with ondansetron and methylprednisolone as above. RESULTS: Analysis of the primary efficacy criterion (absence of emetic episode throughout the course of chemotherapy) revealed a success rate of 53% in the group receiving O + M + MPZ and 38% in the group receiving O + M (P = 0.008). Analysis of the secondary efficacy criteria (nausea grade, number of emetic episodes and global patient satisfaction on D1 and from D2 to D3) showed a statistically significant difference between the two groups, in favour of the O + M + MPZ treatment. The scores obtained with the FLIC and FLIE questionnaires did not reveal any significant differences between the two groups. Treatment was well tolerated in both groups. CONCLUSION: The study showed that the addition of MPZ to the combination O + M was an effective and well tolerated antiemetic treatment, with a 15% increase in efficacy compared to the combination in patients not controlled during their previous course of chemotherapy. The addition of metopimazine to existing regimens containing 5HT3 receptor antagonist and steroid combination should be considered for patients who fail on their previous course. PMID- 9358941 TI - Preliminary report of an intensified, short duration chemotherapy protocol for the treatment of pediatric non-Hodgkin's lymphoma in India. AB - BACKGROUND: In the past, the results of the treatment of non-Hodgkin's lymphomas (NHL) in Indian children have been poor, due to inadequate chemotherapy and poor supportive care. In an attempt to overcome these problems, we conducted a clinical trial in Bombay with a new protocol, MCP842. PATIENTS AND METHODS: Seventy-four previously untreated patients < 25 years were entered on study at the Tata Memorial Hospital, Bombay. Patients with lymphoblastic lymphoma (LL) (38) without bone marrow involvement and all patients with small noncleaved cell lymphoma (SNCL) (18) and large cell lymphoma (LCL) (18) were eligible. Treatment consisted of alternating cycles of two regimens, A and B. Patients with St. Jude stages I and II received six cycles, and those with stages III or IV received eight cycles. A cycles included cyclophosphamide, adriamycin, vincristine and ara C, and B cycles, etoposide, vincristine, methotrexate, ifosfamide and mesna. RESULTS: Complete response was achieved in 67 (91%) of patients. Event free survival (EFS) for all patients was 58%; 68% for patients with SNCL and LCL combined, and 48% for patients with LL. There was no significant difference in EFS by histology (LL versus non-LL), or stage. There were nine (12%) toxic deaths, two during induction and seven in patients in remission; six occurred in patients with LL. CONCLUSIONS: These results are better than past results in Bombay. Unlike earlier CCG protocols, in which the outcome between patients with LL and non-LL differed, this was not so in MCP842. Even patients with extensive LL without bone marrow disease received only eight cycles of therapy, suggesting that short duration therapy is curative in as many as half of such patients--an important observation in a country with limited resources. PMID- 9358942 TI - Risk factors for local recurrences after limb-salvage surgery for high-grade osteosarcoma of the extremities. AB - BACKGROUND: Improvements in preoperative staging as well as in chemotherapeutic regimens have made limb-salvage surgery a reliable modality of treatment for high grade osteosarcomas of the extremities, with local recurrences in most series of less than 10% after this type of surgery. The quality of surgical margins and local response to preoperative chemotherapy are known to be the most significant factors in recurrence [1, 8-10, 12], and complications related to the biopsy procedure may also be a significant factor. The study reported here comprised a histopathological analysis of our recurrent cases as part of an effort to identify the impact of each of the factors cited above. MATERIALS AND METHODS: Five hundred fourteen cases of high-grade, non-multicentric osteosarcoma of the extremities were treated at the Istituto Ortopedico Rizzoli between March 1983 and August 1991. In this study we analyzed 23 cases of local recurrence in patients with classic osteosarcoma who underwent limb-salvage procedures. RESULTS: In 15 cases we found correlation between the site of local recurrence and the site where the margins were less than wide. In five cases the recurrence was secondary to complications of the biopsy procedure (hematoma, delayed healing). In one case we suspect a previously undetected skip lesion. In the remaining two cases no clear explanation was found for the recurrence. There was also a statistically significant difference in the time of appearance of recurrences related to the tumor response to chemotherapy. CONCLUSIONS: For only two cases of recurrence was there no clear explanation. In one we suspect an undetected skip metastasis, and in the other there were certain factors which may have increased its risk of recurrence (non diagnostic trochar biopsy followed by an incisional biopsy, fair tumor necrosis, recurrence in a 'problem' anatomical site, i.e., the popliteal space). In the remaining cases the following factors were found to be directly related to the development of a local recurrence: a) the quality of the surgical margins, b) site of the biopsy as well as complications related to the biopsy procedure, c) local response to preoperative chemotherapy. PMID- 9358943 TI - Pulmonary infiltration associated with myelodysplasia. AB - Four case histories are reported in which the initial signs and symptoms were those of pulmonary infiltration and in which subsequently a diagnosis of myelodysplasia was made. The analysis of bronchoalveolar lavage fluid- demonstrating predominantly neutrophils and lymphocytes, and, occasionally blast cells as well as plasma cells--indicated that the pulmonary infiltration was related to the myelodysplastic process. As no other causes of pulmonary infiltration could be found, it seems that a pulmonary infiltrate can be the presenting symptom of a myelodysplastic syndrome. Although pleuropulmonary infiltrates most often are caused by infections, these cases illustrate that myelodysplasia related infiltrates should also be considered. PMID- 9358944 TI - Concomitant brain radiotherapy and high-dose ifosfamide in brain relapses of lung cancer. AB - PATIENTS AND METHODS: Twenty patients with lung cancer and brain metastasis were prospectively included in this feasibility study (four small-cell and 16 non small-cell lung cancers). There were two previously untreated patients and 18 pretreated patients for whom brain metastases constituted the first relapse after a treatment-free interval following chemotherapy for the primary lung cancer. Most of the patients had neurological symptoms and an ECOG performance index over 2. Treatment consisted of three courses of whole brain radiotherapy (18 Gy in 10 fractions) and ifosfamide: 3 g/m2 daily from day 1 through day 4, i.e., during the first four days of radiotherapy with uromitexan uroprotection and haematopoietic support (r-HuG-CSF). RESULTS: Seventeen patients completed the three-cycle programme. Sixteen patients had grade 4 neutropenia and six of them experienced a febrile episode. Other toxicities were mild to moderate and manageable. The received dose-intensity of ifosfamide was 90%. Response evaluation demonstrated stable disease for two patients, partial response for eight, complete response for six and progression for four. All responders benefited by a remission of symptoms and improvement of performance index. Median survival from start of protocol was 13 months. CONCLUSION: Brain radiotherapy plus high-dose ifosfamide is feasible in patients suffering from brain recurrences of lung cancer. PMID- 9358945 TI - Subcutaneous clodronate: a study evaluating efficacy in hypercalcemia of malignancy and local toxicity. AB - The logistics of administering intravenous bisphosphonates may be problematic in the care of advanced cancer patients, especially in the home setting. Hypodermoclysis is a convenient method of administering fluid via subcutaneous infusion, presently used in the domiciliary setting. Results of the administration of clodronate via this route are reported. PMID- 9358946 TI - Modulation of 5-fluorouracil with methotrexate and low-dose N-(phosphonacetyl)-L aspartate (PALA) is inactive in advanced pancreatic carcinoma. AB - PURPOSE: To evaluate the effect of biochemical modulation by PALA and methotrexate on the therapeutic activity of 5-fluorouracil (5-FU) in patients with advanced pancreatic adenocarcinoma. PATIENTS AND METHODS: The treatment protocol consisted of phosphonacetyl-L-aspartate (PALA) 250 mg/m2 i.v. 15-minute infusion followed by methotrexate 200 mg/m2 i.v. 30-minute infusion on day 1 and 5-FU 600 mg/m2 i.v. push on day 2. Folinic acid was given at 15 mg/m2 p.o. every six hours for eight doses, starting 24 hours after methotrexate infusion. Cycles were repeated every two weeks. RESULTS: Thirty patients with advanced chemotherapynaive pancreatic cancer were included; 26 had measurable disease. Median age 56 years (27-72); median PS 1 (0-2). One PR (3.9%) was achieved; nine patients had stable disease. Median time to progression was 91 days. Median survival was 177 days and one year survival was 13.3% (4 of 30 patients). Treatment was well tolerated; diarrhea WHO grade 2 or 3 occurred in six patients; stomatitis WHO grade 2 and 3 in nine patients. CONCLUSIONS: Modulation of 5-FU by PALA and MTX given in this dose and schedule appears to be ineffective in patients with advanced pancreatic adenocarcinoma. PMID- 9358947 TI - Paraneoplastic temporal lobe epilepsy and anti-Yo autoantibody. PMID- 9358948 TI - Recurrent bowel occlusion with oral ondansetron with no side effects of the intravenous route: a previously unknown adverse event. PMID- 9358949 TI - Dipole localization and test-retest reliability of frequency and duration mismatch negativity generator processes. AB - The mismatch negativity (MMN) is an event related potential component elicited by changes in duration, frequency or intensity of the stimuli during repetitive series of equal standard stimuli. In the present study we compared duration and frequency MMN using dipole source analysis concerning both the test-retest reliability of MMN-amplitudes and the locations of the potential sources. Furthermore, the influence of attention for test-retest-reliability was studied. Therefore, two groups of healthy subjects were investigated with different attentional manipulations. Twenty-one healthy subjects had to perform a visual attention task during the recording and 21 healthy subjects had no additional task to perform. All subjects were studied twice with a time interval of 3 weeks. Test-retest reliability was sufficiently high for the frequency but slightly lower for the duration MMN. The locations of the frequency and duration MMN dipoles were in the auditory cortex with a more anterior and caudal location for the frequency MMN-dipoles. The latter finding supports the hypothesis that the frequency and duration MMNs have separate neuronal generators. PMID- 9358950 TI - Sources and topography of supramodal effects of spatial attention in ERP. AB - This study aimed to finding modality-independent event-related potentials (ERP) modulations related to spatial attention by minimising several possible methodological constrains which may account for the rareness of this finding in past spatial attention ERP research. ERP were recorded while subjects performed a shifting spatial attention task of either auditory, visual or mixed (both auditory and visual) modality. Task requirements and stimulation devices were identical for either modality. Stimuli could appear (80%, valid trials) or not (invalid trials) at predicted locations, and subjects had to perform a speeded response to every stimulus. Reaction time benefits were found for valid trials. Instead of the usually found enhancement of sensory-evoked modality-specific components for valid trials, ERP showed modality-independent modulations as a function of attention. They consisted of a fronto-central activity starting 240 ms after stimulus onset and originated in the anterior cingulate gyrus, followed 40 ms later by a bilateral parietal cortex activation. These processes would be reflecting the activity, respectively, of the anterior and posterior attention systems postulated in Posner's model (Posner and Peterson 1990), which showed a higher degree of participation during invalid trials. P300-like processes were also observed for invalid trials. These results demonstrate that supramodal effects of spatial attention of ERP can be obtained. Furthermore, they extend Posner's model by indicating the timings of implication of the attention subsystems. PMID- 9358952 TI - Confidence interval of single dipole locations based on EEG data. AB - Noise in EEG and MEG measurements leads to inaccurate localizations of the sources. A confidence volume is used to describe the amount of localization error. Previous methods to estimate the confidence volume proved insufficient. Thus a new procedure was introduced and compared with previous ones. As one procedure, Monte Carlo simulations (MCS) were performed. The confidence volume was also estimated using two methods with different assumptions about a linear transfer function between source location and the distribution of the potential. One method used variable (LVM) and the other fixed dipole orientations (LFM). Finally, the confidence volume was estimated through a procedure in which there was no linearization of the transfer function. This procedure scans the confidence volume by varying the dipole location in multiple directions. Confidence volumes were calculated for simulated distributions of the electrical potential and for experimental data including somatosensory evoked responses to stimulation of lower lip, thumb, and little finger. Results from simulated data indicated that confidence volumes calculated with the MCS method were largest, and those calculated with the LFM method were smallest. For dipole locations close to the brain surface, the confidence volume was smaller than for a central deeper source. An increase in electrode density resulted in smaller confidence volumes. When the noise was correlated, only the method using the MCS produced acceptable results. Since the noise in experimental data is highly correlated, only the MCS method would appear to be useful in estimating the size of the confidence volume of the dipole locations. Thus, using real data with the MCS method, we easily distinguished separate and distinct representations of the thumb, little finger, and lower lip in the somatosensory cortex (SI). It was concluded that adequate estimation of confidence volumes is useful for localizing neural activity. On a practical level, this information can be used prior to an experiment for determining the conditions necessary to distinguish between different dipole sources, including the required signal to noise ratio and the minimum electrode density. PMID- 9358951 TI - Activity in posterior parietal cortex following somatosensory stimulation in man: magnetoencephalographic study using spatio-temporal source analysis. AB - We investigated the activation of posterior parietal cortex (PPC) to somatosensory stimulation in humans to determine its fundamental role as a somatosensory associated area using magnetoencephalography (MEG). We studied somatosensory evoked magnetic fields (SEF) after stimulation of median nerve, posterior tibial nerve and lip, and analyzed them by the single dipole model and also by the multidipole model using brain electric source analysis (BESA) system. In single source model analysis, the dipole at the peak latency of short-latency components following each site stimulation were located in the corresponding receptive fields in the primary somatosensory cortex (SI) contralateral to the stimulation. The dipole at the peak latency of the middle latency components were located in bilateral upper bank of Sylvian fissure (SII), By contrast, in the five-dipole model of BESA, the equivalent current dipoles (ECDs) of the middle latency SEF after stimulation of median nerve and posterior tibial nerve were identified in the contralateral SI and in the bilateral SII and PPC, while all activities of middle-latency SEF after lip stimulation appeared to be restricted in the contralateral SI and bilateral SII. Around 80 msec in latency, the ECD location in PPC after median nerve stimulation was, on the average, 2.4 cm posterior, 2.9 cm medial and 2.6 cm superior to the hand area in SI. The ECD in PPC after posterior tibial nerve stimulation was also located posterior to the foot area in SI, but it was close to the SI area of foot, their distance being approximately 1.3 cm. ECD in PPC was almost equally demonstrated in each hemisphere. These findings suggested that the somatosensory associated cortex in PPC represented somatotopic organization in parallel with 'homunculus' in SI, but the hand area was much wider than the foot area. It was not clear whether the lip area in PPC was absent or was too close to be separated from the SI. PMID- 9358953 TI - Methohexital-induced changes in spectral power of neuromagnetic signals: beta augmentation is smaller over the hemisphere containing the epileptogenic focus. AB - Previous research has suggested that methohexital, a short-term barbiturate, alters activity in the primary epileptogenic area. It can be assumed that drug induced activation of the epileptogenic focus provides a rapid and safe method to obtain a sufficient amount of information relevant for the lateralization and localisation of the primary epileptogenic area. This study shows that methohexital changes spectral power in the beta band derived from magnetoencephalographic (MEG) signals over the hemisphere ipsilateral to the primary epileptogenic area. This effect was demonstrated for 10/13 of the investigated patients suffering from unilateral temporal lobe epilepsy (TLE). The side and location of the primary epileptogenic area of these patients (5 left TLE, 8 right TEL) was determined invasively during presurgical evaluation. During a 1-2 minute interval after intravenous bolus injection of 100 mg methohexital a clear lateralization effect in the beta band was observed, which differed marginally between fronto-central, fronto-temporal and temporo-parietal brain regions. In addition, bilateral spectral power changes were obtained in the theta, alpha and gamma bands that differed between brain regions. Analyses of simultaneously recorded scalp electroencephalographic (EEG) data revealed effects consistent with those of the MEG analysis. The reduced enhancement of beta band spectral power of MEG recordings provides a potential application for the non invasive lateralization of the primary epileptogenic area. PMID- 9358954 TI - Combination treatment in antihypertensive drug trials. PMID- 9358955 TI - New developments in cardiovascular drugs: heart failure--an informal and personal viewpoint. PMID- 9358957 TI - Effect of ramipril on heart rate variability in digitalis-treated patients with chronic heart failure. AB - The aim of this study was to evaluate the effect of an angiotensin-converting enzyme (ACE) inhibitor, ramipril, on heart rate variability in patients with heart failure simultaneously treated with digitalis. This study was a multicentric, randomized, double-blind, placebo-controlled study including 50 patients with chronic heart failure (CHF). All patients were in NYHA functional class II and III. The etiology of CHF was mainly idiopathic dilated cardiomyopathy and ischemic heart disease. After a 4-week placebo run-in period with digoxin and diuretics, patients were randomized to receive additional ramipril or placebo. To assess heart rate variability (HRV) and arrhythmias, 24 hour ECGs were recorded at the end of the placebo run-in period, 8 and 24 weeks after randomization. Spectral analysis of HRV was performed during one diurnal and one nocturnal 5-minute time period. No statistically significant differences in HRV within low-, high-, and total-frequency bands were induced by ramipril in either the diurnal or nocturnal periods, both at 8 and 24 weeks after randomization. Ramipril produced a significant decrease in nonsustained ventricular tachycardia at 24 weeks of treatment (p = 0.01). These results run against previous observations showing an increase in parasympathetic tone with ACE inhibitors in heart failure. The present study thus suggests that the effects of ACE inhibitors in CHF are variable and depend on the patient and concomitant treatment that might influence HRV such as digoxin treatment. PMID- 9358956 TI - Angiotensin II receptor blockade in Syrian hamster (T0-2) cardiomyopathy does not affect microscopic cardiac material properties: implications for mechanisms of tissue remodeling. AB - This study delineates the role of angiotensin II type I (AT1) receptor in the remodeling of Syrian cardiomyopathic hamsters. Twelve cardiomyopathic (T0-2) hamsters received L-158,809 treatment and libitum in their drinking water (27 micrograms/ml) and 9 cardiomyopathic and 9 normal FL-B hamsters received tap water from 1 to 4 months of age. Although pharmacologically effective with regard to complete suppression of the blood pressure response to angiotensin II infusion, L-158,809 did not diminish the progression or severity of cardiomyopathy. Heart weight/100 g body weight and left ventricular wall thickness adjusted for body weight of both L-158,809 and cardiomyopathic control hamsters did not differ and exceeded those of F1-B controls (p < 0.05). Myocardial material properties (e.g., stiffness and density) of cardiomyopathic hamsters treated with L-158,809 were not affected. Thus, the progression of fibrosis, calcification, and necrosis in T0-2 cardiomyopathic hamsters was not sensitive to AT1 receptor blockade. PMID- 9358958 TI - Cardiovascular actions of the dopamine receptor agonist Z1046 in swine. AB - Dopamine receptor agonists can be useful in the treatment of hypertension or heart failure. Consequently, the present study investigated the hemodynamic profile of the novel dopamine D1/D2 receptor agonist Z1046 in open-chest, pentobarbital-anesthetized swine. Z1046 was administered in a dose of 10 micrograms/kg (n = 9) or 100 micrograms/kg (n = 8), which was injected over 1 minute; hemodynamic responses were studied for 90 minutes after administration. Both doses of Z1046 produced sustained decreases in mean aortic blood pressure (15-20%). The hypotension produced by the lower dose was principally due to a decrease in cardiac output, as the trend toward a lower systemic vascular resistance failed to reach levels of statistical significance. Conversely, the decrease in mean arterial blood pressure produced by the higher dose of Z1046 was mainly due to a decrease in systemic vascular resistance (up to 17%), as the trend toward a decrease in cardiac output at 60 and 90 minutes after administration was not different from the changes in the saline-treated group. Heart rate decreased slightly with both doses of Z1046. Z1046 decreased left ventricular myocardial blood flow (up to 28 +/- 9%, p < 0.05) in parallel with the decrease in myocardial oxygen consumption (up to 24 +/- 7%, p < 0.05), with no change in the transmural distribution of myocardial blood. Z1046 in a dose of 10 micrograms/kg did not produce significant vasodilation of regional vascular beds, but in a dose of 100 micrograms/kg produced vasodilator responses in the small intestine (34 +/- 2% decrease in vascular resistance), spleen (43 +/- 7%), and kidneys (22 +/- 3% all p < 0.05 vs. baseline). In conclusion, Z1046 produced systemic hypotension with negligible reflex activation of sympathetic tone. PMID- 9358959 TI - Effects of protease inhibitors on postischemic recovery of the heart. AB - It is well known that activation of proteases in the lysosomes and cytosol is one of the mechanisms of ischemic injury. It might thus be beneficial to determine whether the addition of several clinically available protease inhibitors to a cardioplegic solution can improve its protective ability. Using an isolated working rat heart preparation, the effects of several protease inhibitors (serine protease inhibitors; nafamostat mesilate and gabexate mesilate, a thiol-protease inhibitor; NCO-700; and a urinary trypsin inhibitor, urinastatin) on the postischemic recovery of function and enzyme leakage were investigated in this study. These protease inhibitors were added to either the cardioplegic solution or reperfusion solution. The addition of each of the protease inhibitors, except urinastatin, to the cardioplegic solution improved the postischemic recovery of function and reduced enzyme leakage. The dose-response characteristics of these three protease inhibitors were bell shaped, and the optimal concentrations of nafamostat mesilate, gabexate mesilate, and NCO-700 were 5 microM, 100 microM, and 20 microM, respectively. In contrast to the results of the preischemic treatment study, the addition of any of the protease inhibitors to the perfusion medium during Langendorff reperfusion failed to improve the postischemic recovery of function and to reduce enzyme leakage. Surprisingly, the addition of NCO-700 to the reperfusion solution at a concentration of 5 microM or higher had rather harmful effects on both functional recovery and enzyme leakage. These findings suggest that serine and thiol proteases may play an important role in myocardial injury during ischemia, but not necessarily during reperfusion. PMID- 9358960 TI - Infarct size as estimated from peak creatine kinase and lactate dehydrogenase is probably reduced in patients using calcium antagonists at the onset of symptoms. AB - In animal models, calcium antagonists (Ca-A) administered before ischemia and reperfusion reduced myocardial necrosis, attenuated postischemic contractile dysfunction, and reduced tissue calcium. In 753 patients with acute myocardial infarction (AMI), we examined if use of Ca-A at the onset of symptoms (n = 127 patients) reduced infarct size as estimated from peak creatine kinase (CKmax) and lactate dehydrogenase (LDmax) activities. The study had an observational exposed/nonexposed design, and both crude and adjusted effects were investigated. Crude effects: In the restricted cohort of patients not receiving thrombolytic treatment (thr- pts; n = 411 patients), CKmax and LDmax were lower in Ca-A+ patients than in Ca-A- patients, being 643 versus 887 U/l (2 p = 0.004) and 708 versus 867 U/l (2 p = 0.005), respectively. When using log (CKmax) and log (LKmax) as outcomes, the same results were found (2 p = 0.002). More of the restricted cohort of the pts used Ca-A in the lower quartiles of CKmax and LDmax (p for linear trend = 0.005 and 0.004 for CKmax and LDmax, respectively). Adjusted effects: Thrombolysis was an effect modifier of the association between Ca-A and peak enzyme levels. In thr-pts, the coefficients of Ca-A were negative and borderline significant for log (CKmax; 2 p = 0.088) and negative and highly significant for log (LDmax; 2 p = 0.010) when adjusting for confounders. The present observational study indicates that the use of a Ca-A at the onset of AMI reduces infarct size, as estimated from CKmax and LDmax activities. PMID- 9358962 TI - Antioxidant effects of lovastatin and vitamin E on experimental atherosclerosis in rabbits. AB - The effects of the administration of vitamin E (10 mg/day) plus lovastatin (2 mg/day; group A, n = 10), lovastatin alone (2 mg/day; group B, n = 10), and placebo (group C, n = 10) were compared over 24 weeks in a randomized, single blind controlled trial. All groups of rabbits received a trans fatty acid (TFA) rich diet (5-10 g/day) for 36 weeks. Treatment with vitamin E plus lovastatin (group A) and lovastatin (group B) started after 12 weeks of administration of TFA-rich diet was associated with a significant but similar decline in serum cholesterol, low-density lipoprotein (LDL) cholesterol, and triglycerides in both groups at 36 weeks. Lipid peroxides and diene conjugates showed a significant decline in association with a significant increase in the plasma level of vitamin E in group A rabbits at 36 weeks. However, the lovastatin group B showed a lesser but significant decrease in lipid peroxides and diene conjugates at 36 weeks, indicating that lovastatin may have antioxidant activity. In control group C, the increase in blood lipids and oxidative stress at 36 weeks was much greater than the decrease in groups A and B. After experimental lipid peroxidation at 24 weeks in all of the rabbits, 2 of 10 group B and 3 of 10 group C rabbits died due to coronary thrombosis; there were no deaths in group A. Thus antioxidant therapy with vitamin E can provide protection against death due to free radical stress. Aortic lipids and sudanophilia indicating athorosclorosis were significantly lower in groups A and B than in group C. The atherosclerotic coronary plaque sizes were significantly smaller in group A (18.5 +/- 3.6 microns) than in groups B (41.6 +/- 4.2 microns) and C (85 +/- 6.7 microns). Aortic plaque sizes were also smaller in group A than in group B and C. It is possible that antioxidant therapy with vitamin E, as an adjunct to lipid lowering with lovastatin, can provide additional benefit in the inhibition of oxidative stress and atherosclerosis. The antioxidant activity of lovastatin has not been reported, to our knowledge. PMID- 9358961 TI - Reduction of plasma cholesterol levels and induction of hepatic LDL receptor by cerivastatin sodium (CAS 143201-11-0, BAY w 6228), a new inhibitor of 3-hydroxy-3 methylglutaryl coenzyme A reductase, in dogs. AB - The effects of cerivastatin sodium (BAY w 6228), a new type of inhibitor of 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, on plasma cholesterol concentrations and the induction of hepatic LDL receptors were investigated with beagle dogs and Hep G2 cells. Oral administration of cerivastatin (0.01, 0.03, and 0.1 mg/kg per day) for 3 weeks reduced plasma total and very low-density lipoprotein plus low-density lipoprotein (VLDL + LDL) cholesterol concentrations and increased hepatic LDL receptor binding activity in dogs. Scatchard plot analysis revealed a 1.9-fold increase in the maximum binding capacity of hepatic LDL receptors in cerivastatin-treated animals. Similar results were obtained by administration of pravastatin (1.0 and 5.0 mg/kg/day) for 3 weeks. Binding activity of the LDL receptor, as well as receptor mRNA and protein concentrations, were increased in a dose-dependent manner (0.01-1.0 microM) by exposure of Hep G2 cells to cerivastatin. The results suggest that cerivastatin reduces plasma cholesterol concentrations by increasing hepatic LDL receptor expression. The mechanism of lowering cholesterol concentration by cerivastatin was the same as with the other previously examined HMG-CoA reductase inhibitors, but the effects with cerivastatin were apparent at doses much lower than the effective doses of the other drugs. Cerivastatin, therefore, shows potential for clinical use as a potent and efficacious plasma cholesterol-lowering drug. PMID- 9358963 TI - Nisoldipine CC and lisinopril alone or in combination for treatment of mild to moderate systemic hypertension. Canadian Nisoldipine CC Hypertension Trial Group. AB - The efficacy and safety of nisoldipine CC and lisinopril were compared in 278 patients with mild to moderate systemic hypertension in a double-blind, placebo run-in trial. Patients were randomized to nisoldipine CC or lisinopril for 8 weeks to achieve a trough sitting diastolic blood pressure (BP) < or = 90 mmHg. Responders were maintained on their optimal dose for a further 8 weeks. Nonresponders were switched to combination therapy and treated for 8 weeks. Twenty-four-hour ambulatory BP monitoring (ABPM) was carried out during placebo and monotherapy. The responder rate of 73.8% with nisoldipine CC after 8 weeks was greater than 56.1% with lisinopril (p = 0.007). The responder rate with combination therapy was 61%. ABPM showed that both nisoldipine CC and lisinopril produced constant blood pressure lowering effects over the 24-hour period and maintained circadian rhythm. Adverse effects were more frequent with nisoldipine CC (headache and peripheral edema) than with lisinopril (cough) monotherapy. Nisoldipine CC monotherapy was at least as effective as lisinopril monotherapy in the management of mild to moderate hypertension. Both agents were well tolerated. Combination therapy with nisoldipine CC and lisinopril was effective and well tolerated in patients with blood pressure not controlled by monotherapy alone. PMID- 9358964 TI - Effects of azimilide and d,l-sotalol on the heart rate and blood pressure response to isoproterenol in anesthetized rats. AB - The novel class III antiarrhythmic agent, azimilide, provides antifibrillatory protection in a rat model of ischemia-reperfusion arrhythmias. In other species azimilide's antifibrillatory mechanism is thought to be mediated predominantly through blockade of both the rapid and slow components of the delayed rectifier potassium current in ventricular myocytes. However, the delayed rectifier potassium current does not appear to control cardiac repolarization in the rat. One possible mechanism for antiarrhythmic efficacy in rats is the compound's beta adrenergic blocking effect, previously seen in isolated guinea pig hearts. The purpose of this study was to evaluate the beta-adrenergic antagonistic effect of azimilide in the rat. Beta-adrenergic blockade was evaluated in the intact anesthetized rat by studying the effects of intravenous azimilide (at or above the antifibrillatory dose) and d,l-sotalol (a known beta-adrenergic antagonist) on heart rate and blood pressure responses to isoproterenol (0.14 Mg/kg i.v.). d,l-Sotalol (6.0 mg/kg) reduced (p < 0.05) the tachycardic response to isoproterenol from 133 +/- 11 to 80 +/- 10 beats/min, and 3.0 mg/kg of d,l sotalol reduced the hypotensive response from -74 +/- 4 to -43 +/- 5 mmHg. Azimilide (5.0, 10.0, and 20.0 mg/kg) did not have a statistically significant effect on either the heart rate or blood pressure changes caused by isoproterenol. These data demonstrate that azimilide does not have a beta adrenergic antagonist effect in the rat at antifibrillatory doses. Therefore, the antiarrhythmic effect of azimilide in the rat is mediated through a mechanism other than beta-blockade. PMID- 9358965 TI - Loading doses of amlodipine safely increase plasma levels during the first days of treatment in patients with angina pectoris. PMID- 9358966 TI - Migraine without aura and ACE-gene deletion polymorphism: is there a correlation? Preliminary findings. PMID- 9358967 TI - Neurovascular relationships in the posterior cranial fossa, with special reference to trigeminal neuralgia. 1. Review of the literature and development of a new method of vascular injection-filling in cadaveric controls. AB - Vascular compression of cranial nerves adjacent to the brain stem has been implicated in a wide variety of disorders affecting their function. The considerable conflicts in published results relate primarily to flaws in study design. The design required of an adequate study is defined and a technique is presented, in 16 fresh human cadavers, of reliable and physiological injection filling of both the cerebral arterial and venous systems. It allowed for the accurate observation of the normal neurovascular relationships in the posterior cranial fossa during operative simulation. Part 2 of this article concerns the use of this design in the study of trigeminal neuralgia, a disorder thought to relate to vascular compression of the fifth cranial nerve. PMID- 9358968 TI - Neurovascular relationships in the posterior cranial fossa, with special reference to trigeminal neuralgia. 2. Neurovascular compression of the trigeminal nerve in cadaveric controls and patients with trigeminal neuralgia: quantification and influence of method. AB - The theory of neurovascular compression has been tested by comparing the neurovascular relationships of the trigeminal nerve in a series of operative observations in patients affected by trigeminal neuralgia with those of a control series of cadavers matched for age, sex and side, in which operative conditions were simulated during simultaneous arterial and venous injection--filling to physiological pressures, as described in Part 1 of this article. A rigorous system of classification of neurovascular relations is defined. In 46 patients with trigeminal neuralgia, 91% had a vessel in contact with the trigeminal nerve adjacent to the brain stem and in all but one a groove was created. Multiple vessels were found in 17% and in two both the root entry zone and lateral portions of the nerve were compressed. However, in 35 randomly selected fresh cadavers, not known to have suffered neurological disease, 14% had neurovascular contact and a further 26% had vessels "near" to the nerve. No vessel was associated with a groove and no multiple vessels, or sites of contact, were encountered. The difference between the control cadavers and the operative findings in patients related to an increase in the number of arteries. Injection filling of the cadaveric vessels doubled the numbers of vessels in contact with, and near to the nerve. The technique used and system of classification applied showed an association between arterial contact and trigeminal neuralgia. The technique may provide a suitable method for the testing of the neurovascular compression theory in other conditions. PMID- 9358969 TI - Anomalous branching order of the superior and lateral thoracic arteries. AB - The superior thoracic and lateral thoracic arteries usually arise from the axillary artery as the first and second branches, respectively. Although anomalies have been described, no reports of a reversal of the order of origin have been found In two cadaveric dissections, the lateral thoracic artery was found to arise cephalad to the superior thoracic artery. PMID- 9358970 TI - Anatomy and magnetic resonance imaging of the posterolateral structures of the knee. AB - The purpose of the present study was to provide detailed information of the morphological and radiological characteristics of the posterolateral structures of the knee. Muscles and ligaments of the posterolateral part of the knee were studied by dissections of 50 adult cadaver knees and by Magnetic Resonance Imaging (MRI) before and after dissections for comparisons. Diverse morphological characteristics of the arcuate ligament were found. The fabellofibular ligament was present in 42.1% of the knees dissected, whereas the popliteofibular ligament was found in 37.5%. A ligamentous structure, which could be called the posterior tibial ligament, was found in 31.6% of the cases that originated from the lateral part of the capsule proximally and inserted distally on the mid portion of the proximal tibia. By comparing the cross sections and the dissections of the cadaver knees, the popliteus muscle, the arcuate ligament, the fibular collateral ligament, the popliteofibular ligament, and the fabellofibular ligament could be identified in MRI. Comprehensive understanding of the posterolateral anatomy of the knee and improved identification of the structures in MRI will help clinicians to make a more accurate and noninvasive diagnosis of posterolateral instability. PMID- 9358971 TI - CT anatomy of the mediastinal structures at the level of the manubriosternal angle. AB - The mediastinal structures said to lie on a horizontal plane at the level of the manubriosternal joint (manubriosternal plane) in the cadaver include the bifurcation of the trachea, the concavity of the arch of the aorta, and the azygos vein as it arches over the right principal bronchus to enter the superior vena cava. We have reviewed CT scans of the thorax in 51 subjects to determine 1) whether these structures lie consistently at this level in the living thorax and 2) the vertebral level of this plane. We found that the bifurcation of the trachea lay at the plane in 41% of subjects, that the plane passed through the concavity of the arch of the aorta in 49% of subjects, and that, although there was notable individual variation, the manubriosternal plane passed through the upper part of the fifth thoracic vertebra in 53% of cases. PMID- 9358973 TI - Mandibular spine: a case report. AB - This is a brief review of the origin and normal variations of the genial tubercles, and specifically, a mandibular spine as an anatomical variation or abnormality. The case presented is that of abnormal genial tubercles observed during dissection of the floor of the mouth. A review and description on the morphology of the genial tubercles are given. PMID- 9358972 TI - Trigeminal neuralgia: an anatomically oriented review. AB - Trigeminal neuralgia (TGN) is a peculiarly painful paroxysmic disorder with an annual incidence of 4.3 per 100,000, which undergoes spontaneous remissions and recurrences. The pain, which is subserved by large not small fibers, can in some cases be triggered from outside the trigeminal territory and by other than mechanical stimuli. There are strong autonomic influences on the pain, and there is cutaneous vasoconstriction in the trigeminal territory in which it occurs. There are also sensory perception deficits for temperature in the affected region and for touch in the whole trigeminal territory. There is now increasing evidence that the majority of cases are caused by vascular compression of the fifth nerve at its point of entry into the pons, for the pain can be relieved (with restoration of the sensory deficit) by surgical decompression. No anatomical abnormalities of the (peripheral) trigeminal nerve have ever been satisfactorily demonstrated. Arguments are examined for the hypothesis that TGN is essentially a disorder of central processing, the term being taken to include the oligodendroglial-sheathed proximal segment of the nerve. PMID- 9358975 TI - Pathophysiology of central nervous system complications in diabetes mellitus. AB - It is now generally accepted that diabetes can alter central nervous system (CNS) function. Even in the absence of overt cerebrovascular accidents or repeated hypoglycemic reactions, uncontrolled hyperglycemia is associated with cognitive changes. These changes are documented both in patients with diabetes as well as in animal models of experimental diabetes. The cognitive impairment can be ameliorated with optimization of blood glucose control. The potential causes of CNS dysfunction in diabetes can be broadly categorized as either vascular causes including changes in the blood-brain barrier and metabolic changes. The latter causes include repeated hypoglycemic episodes, hyperglycemia, hyperosmolality, acidosis, ketosis, neuroendocrine or neurochemical changes. The other contributory causes of CNS dysfunction in diabetes include the presence of hypertension, uremia, peripheral and autonomic neuropathy and multiple drug use. PMID- 9358974 TI - Brachial plexus palsy and spinal subarachnoid pneumatosis following trauma. AB - The case of a 33-year-old male involved in a road traffic accident, resulting in brachial plexus palsy associated with subarachnoid pneumatosis, is described. A knowledge of the regional anatomy at the root of the neck is used to explain the patient's neurological signs and the mechanism of entry of air into the subarachnoid space. PMID- 9358976 TI - Glucose concentration and retinal function. AB - The rod and cone systems of the mammalian retina differ in their structure and functional properties as well as in their metabolic characteristics. This article summarizes basic observations on retinal glucose metabolism reflected in retinal electrophysiology. Metabolic factors might be related to the complex pathogenesis of diabetic retinopathy. Effects of changing glucose concentration and, independently, of insulin on retinal responses obtained in an isolated mammalian eye preparation in vitro and also in vivo are presented. Electron microscopy (EM) histochemical data reveal a distinctive distribution of glycogen in glia and in various subclasses of neurons in the cat retina. Low glucose, corresponding to hypoglycemia in vivo, affected the light-evoked electrical responses from the rod system, but not from the cone system in vitro. This could be confirmed in the anesthetized cat under glucose clamp conditions. Insulin had no influence on physiological retinal function, except under conditions of low glucose, where it enhanced the reduction in b-wave amplitude. This effect is interpreted as a sign of increased glucose utilization by the retinal Muller (glial) cells. PMID- 9358977 TI - Glucose availability and the electrophysiology of the human visual system. AB - Glucose is a main energy source to neurons in brain (with limited storage capability) and retina (where it is stored in glial Muller cells and supplied upon demand). Glucose availability and visual function are related. Human positron emission tomography studies indicate increased blood flow and glucose metabolic rate in primary visual cortex during stimulation, with retinotopic distribution. Retinal electrophysiology covaries with glucose concentration in in vitro models as well as in humans, at comparable concentrations in the physiological range. The interactions between retinal electrophysiology (notably the electroretinogram b-wave) and glucose metabolism appear more stringent than for cortical evoked responses. K-channels regulated by intracellular ATP are thought to link neuron excitability (and electrophysiological activity) on the metabolic state. High-affinity sulphonylurea binding sites for K-channels are widely distributed in brain. K-channels conceivably modulate neurotransmitter release and are inactivated by elevated glucose concentrations and sulfonylurea drugs used to treat diabetes. PMID- 9358978 TI - Clinical features and investigation of diabetic somatic peripheral neuropathy. AB - A multiplicity of peripheral nerve syndromes may develop in patients with diabetes mellitus, the commonest of which is a chronic symmetric sensory polyneuropathy, often associated with autonomic neuropathy. Once established, it is largely irreversible. Acute painful diabetic sensory neuropathy is a separate entity with a favorable prognosis. It now seems likely that chronic inflammatory demyelinating polyneuropathy occurs with greater frequency in diabetic subjects than in the general population and is one explanation for the occurrence of a predominantly motor polyneuropathy. Focal and multifocal peripheral nerve lesions are seen mainly in older diabetic patients and comprise cranial, thoracoabdominal and limb nerve lesions, the last including proximal lower limb diabetic motor neuropathy (diabetic amyotrophy). With this wide array of disorders and the frequency of diabetes, it is important to distinguish those that are directly or indirectly related to diabetes from those that have a coincidental relationship. PMID- 9358979 TI - Autonomic neuropathy: clinical and instrumental findings. AB - The development of sensitive techniques evaluating functions under autonomic control has allowed the early detection of widespread abnormalities in diabetes mellitus. However, despite a high frequency of functional abnormalities, an overt clinical syndrome develops slowly and is quite rare. Characteristic clinical features, more recent methods for evaluating autonomic function, diagnostic procedures, and main instrumental findings in a diabetic population are reported. Emphasis is given to more promising techniques evaluating autonomic control of the cardiovascular system, such as myocardial scintigraphy and assessment of 24-h blood pressure and heart rate variability. The clinical meaning of the number of functional abnormalities observed in diabetic patients is considered. While the role of autonomic neuropathy in the pathogenesis of gastrointestinal motor disorders, hypoglycaemia unawareness or diabetic impotence needs to be revised, the importance of autonomic-related sweating and blood flow abnormalities in the pathogenesis of diabetic foot lesions is now better documented. Moreover, growing evidence of the importance of autonomic control of cardiovascular system, together with cardiovascular dysfunction linked to diabetic autonomic neuropathy, supports the hypothesis of a possible role of autonomic neuropathy in the increased cardiovascular morbidity and mortality observed in diabetic patients. PMID- 9358980 TI - Neuropathology of diabetic neuropathy and its correlations with neurophysiology. AB - Although the detailed pathogenesis of diabetic polyneuropathy is not known, several mechanisms appear to be involved and may occur sequentially. Hence, the early and much researched activation of the polyol-pathway appears to secondarily affect nonenzymatic glycation, perturbation of vasoactive substances, the immune system and neurotrophism. These metabolic abnormalities may be differentially expressed in the neuropathy occurring in insulin dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM) diabetes. This notion is supported by differences in the structural abnormalities of the neuropathies in the two types of diabetes. Distinct and characteristic nodal changes occur in IDDM but not in NIDDM neuropathy, which also shows a milder axonal atrophy. On the other hand, nerve fiber loss which characterizes diabetic neuropathy tends to be focal in the older NIDDM patients, suggesting a more prominent vascular genesis. A further characteristic feature of diabetic neuropathy is blunted fiber regeneration, which probably is consequent to impairments of the necessary immune response and local synthesis of neurotrophic factors. Nerve biopsies from diabetic patients, although not necessary for diagnosis, provide valuable tissue for biochemical and molecular analysis of underlying mechanisms, the detailed elucidation of which will facilitate the design of targeted therapies. PMID- 9358981 TI - Pathogenesis of diabetic neuropathy. AB - Diabetes mellitus affects over 14 million people in the United States and the number of diabetics is increasing by 5% per year. Diabetic neuropathy (DN) is a common complication of diabetes and occurs in approximately 50% of diabetic patients over time. Clinical trials have proven that hyperglycemia almost certainly conditions the development of DN. Despite this fact, we still do not understand the mechanism(s) underlying DN. Several possible etiologies have been proposed including altered metabolism of polyol, lipids, or amino acids, vascular insufficiency, increased superoxide-induced free radical formation, impaired axonal transport or reduced neurotrophism. Accumulating evidence suggests that these defects are likely interrelated and that their interaction(s) within the diabetic milieu are responsible for the development and progression of DN. In this review we will discuss these theories, their interrelationships and how, collectively, these ideas may begin to explain the etiology of DN. PMID- 9358983 TI - Evoked potentials in diabetes mellitus. AB - Abnormalities of central afferent and efferent pathways have been revealed by evoked potential studies in diabetic patients. Central conduction time is only slightly prolonged; in afferent pathways the primary sensory neuron is more affected than in the subsequent stages, probably as an expression of a central peripheral distal axonopathy. Central nervous system abnormalities are more frequent in patients with peripheral neuropathy, but evoked potential can be abnormal even in patients without neuropathy. Brainstem auditory evoked potential (BAEP), somatosensory evoked potentials (SEPs) and visual evoked potentials (VEPs) can be affected together, but isolated abnormalities are more frequently observed. Diffuse neuropathological changes have been found in the optic nerves, periventricular regions, brainstem and spinal cord in postmortem pathological studies. Similar changes have been found in animals with experimental diabetes. The pathophysiology of central nervous system (CNS) abnormalities is uncertain, many causes are probably active in including neural damage: chronic hyperglycemia, hypoglycemic episodes, angiopathy, blood-brain barrier dysfunction and others, still unknown. PMID- 9358982 TI - Autoimmunity and diabetic neuropathy. AB - Autoimmunity has been implicated in the pathogenesis of diabetic neuropathy. A number of putative target antigens have been investigated in the attempt to find a relationship between the presence of autoantibodies and the presence of neuropathy. Attention has been given to antibodies against nerve growth factor, adrenal medulla, sympathetic and parasympathetic structures, glutamic acid decarboxylase and phospholipids. Data in the literature do not support a clear cut difference in frequency of positivity or titres between patients with or without diabetic neuropathy. Moreover, it is not clear whether autoantibodies to the above-mentioned antigens, whenever present, play a role in causing nerve damage or if they simply reflect the presence of nerve damage. Longitudinal studies are needed to clarify some conflicting data in the literature. PMID- 9358984 TI - Does neuropathy develop in animal models? AB - Defects of the peripheral nervous system are common in patients with diabetes mellitus. At least 50% of diabetic patients will develop a form of diabetic neuropathy within 25 years after diagnosis. Currently the cornerstone of treatment lies with the maintenance of euglycaemia using insulin, which has inherent problems of its own. In addition, the signs and symptoms of diabetic neuropathy are often intractable. Therefore, the development of effective treatments for diabetic neuropathy is urgently needed. Thus, animal models have been developed to investigate the pathogenesis of diabetic neuropathy and evaluate potential therapeutic agents. However, no model is perfect and no one would suggest that diabetic rats can replicate the human condition fully. In this review the appropriateness of established animal models of diabetic neuropathy is discussed with reference to the pathology and pathophysiology of the human case with the hope of addresssing some of the questions surrounding this general issue. PMID- 9358985 TI - Pulmonary complications of recreational drug usage. PMID- 9358990 TI - Cigaret smoking and the lung. PMID- 9358991 TI - Allografting for chronic myeloid leukemia. AB - Allogeneic stem cell transplantation is the only curative therapy for chronic myeloid leukemia. There have been several recent advances in this field. Early data suggest that blood-derived stem cells are an effective substitute for bone marrow. Allografting can be performed using progenitor cells from other family members, unrelated donors, and umbilical cord blood. Evidence of early relapse can be detected by assays for the fusion gene, BCR-ABL, using the polymerase chain reaction. Most patients who relapse following allogeneic stem cell transplantation can be treated effectively by transfusion of lymphocytes from their original donors. PMID- 9358989 TI - The effects of opiates on the lung. PMID- 9358986 TI - Respiratory toxicities from stimulant use. PMID- 9358992 TI - Hematopoietic stem cell transplantation for sickle cell anemia. AB - Hematopoietic stem cell transplantation is the only therapy able to cure sickle cell anemia at the present time. So far, transplantations have been undertaken in approximatively 140 sickle cell patients all over the world, with good results. The selection of patients for transplantation remains a subject of dilemma because of the unpredictable course of the disease and the lack of valuable prognostic markers. The selection criteria accepted so far concern young patients under the age of 16, with a morbid course of the disease and having a HLA compatible sibling. In Belgium, patients going back to their country of origin were also considered for transplantation. For 100 patients who underwent transplantation in Europe, the current Kaplan-Meier estimates of overall survival, event-free survival, and disease-free survival rates are 90%, 79%, and 81%, respectively. Benefits and side effects are analyzed. PMID- 9358987 TI - Marijuana. Respiratory tract effects. AB - Daily marijuana smoking has been clearly shown to have adverse effects on pulmonary function and produce respiratory symptomatology (cough, wheeze, and sputum production) similar to that of tobacco smokers. Based on the tobacco experience, decrements in pulmonary function may be predictive of the future development of chronic obstructive pulmonary disease (COPD). However, in the absence of alpha-1-antitrypsin deficiency, the habitual marijuana-only smoker would likely have to smoke 4-5 joints per day for a span of at least 30 yr in order to develop overt manifestations of COPD. The mutagenic/carcinogenic properties of marijuana smoke are also well-established. The potential for induction of laryngeal, oropharyngeal, and possibly bronchogenic carcinoma from marijuana has been documented by several case reports and observational series. Despite this, a relative risk ratio for the development of these tumors has not yet been quantified. Based on a higher frequency of case reports for upper airway cancer compared to bronchogenic carcinoma, marijuana smoking may have a more deleterious effect on the upper respiratory tract. However, this hypothesis remains speculative at best, pending confirmation by longitudinal studies. PMID- 9358993 TI - High-dose chemotherapy with autologous stem cell support for breast cancer. AB - High-dose chemotherapy is increasingly utilized for the treatment of metastatic and high-risk breast cancer. Autologous Blood and Marrow Transplant Registry data reveal that a single high-dose cycle of chemotherapy with stem cell support can produce high complete response rates, with 10% to 20% of patients disease-free at 5 years. For patients with locally advanced disease, pooled results estimate 63% without relapse at 3 years for stage II and III disease and 42% for inflammatory breast cancer. New strategies to improve on these results include the incorporation of newly identified active drugs into established regimens and the use of multiple high-dose cycles. Multicycle chemotherapy produces complete response rates ranging from 35% to 93% depending on the regimen. Additional strategies include the use of monoclonal antibodies, immunotherapy, and gene therapy for post-transplant consolidation. The strongest predictor of long-term disease-free survival in virtually all studies is a complete response prior to or following high-dose chemotherapy. Randomized studies are now in the process of being completed in Europe and the United States with reporting scheduled over the next several years. PMID- 9358988 TI - Volatile substance abuse. PMID- 9358994 TI - Hemopoietic blood and marrow transplants in the treatment of severe autoimmune disease. AB - The past year of activities represents virtually the first year of activities in hemopoietic blood and marrow transplants in the treatment of severe autoimmune disease. The concept of profound hematoimmunoablation followed by blood or marrow transplants for severe autoimmune disease is not new, but the first publications of such cases treated for autoimmune disease alone occurred only in late 1996. Other case reports followed, and the activity has expanded rapidly such that over 40 treated patients have been described, 35 of whom have been entered into a recently created European League Against Rheumatism/European Group for Blood and Marrow Transplantation database (now extended to include data from Australasia and the United States). Four international meetings have been held, the published proceedings of which have formed the basis for an ongoing international collaborative effort. Although very early, some trends have been observed: the majority of patients (mostly with multiple sclerosis or scleroderma) have clinically improved or stabilized; the toxicity of the procedure (mostly autologous bone marrow transplantation) was as observed previously in other disorders; and better results are suggested in patients achieving significant T cell depletion, either through conditioning, graft purging, or both. These impressions must be confirmed by prospective comparative studies using a limited number of regimens. PMID- 9358996 TI - Late effects after allogeneic bone marrow transplantation. AB - Late effects are delayed side effects that can occur months or years after bone marrow transplantation. Late effects become a matter of concern as the number of surviving patients and the length of survival time increase. Late effects express themselves as structural or functional impairment of organs or tissues or as neoplastic growth secondary to the primary treatment. Non-neoplastic late effects affect growth and development in children; endocrine and reproductive function; and the function of the eyes, lungs, kidneys, and other organs. Secondary neoplasms are malignant lymphomas and leukemias that occur early after transplantation. Solid tumors develop later and primarily involve skin and mucous membranes. Intensive chemotherapy and radiotherapy and chronic graft-versus-host disease and its immunosuppressive treatment are potential risk factors. Psychosocial rehabilitation is incomplete in a proportion of these patients. Further investigation is necessary for improvement in the prophylaxis and treatment of late effects. PMID- 9358995 TI - An update from the International Bone Marrow Transplant Registry and the Autologous Blood and Marrow Transplant Registry on current activity in hematopoietic stem cell transplantation. AB - The International Bone Marrow Transplant Registry (IBMTR) and the Autologous Blood and Marrow Transplant Registry (ABMTR) are international research organizations that collect and analyze data on recipients of allogeneic and autologous hematopoietic stem cell transplants, respectively. Over 300 institutions in 47 countries contribute data to the IBMTR and over 150 institutions in North and South America contribute data to the ABMTR. The IBMTR and ABMTR database have information on more than 65,000 transplant recipients, about 40% of allogeneic transplantations performed since 1964, and about half of all autotransplantations in North and South America since 1989. Current transplant activity, as reported to the IBMTR and ABMTR, is summarized in this article as well as key findings from recent IBMTR and ABMTR studies. PMID- 9358997 TI - Unrelated marrow donor registries. AB - Although large registries of volunteer, unrelated marrow donors are now operating around the world, potential recipients are frequently unable to identify suitable donors. These searching patients may benefit from 1) improved international coordination and cooperation; 2) a better understanding of the HLA system and the requirements for HLA matching; 3) improved methods for acquisition, storage, and manipulation of HLA-typing data; and 4) the development of alternative hematopoietic stem cell sources. PMID- 9358998 TI - Cord blood stem cells. AB - Use of bone marrow transplant therapy has been limited both by the availability of suitable stem cell donors as well as the various immunologic complications, namely graft-versus-host disease (GVHD) and profound immunosuppression resulting in opportunistic infection. As of March 1997, more than 450 patients with various malignant and nonmalignant diseases have been transplanted with umbilical cord blood from related and unrelated donors. Preliminary results suggest that rate of engraftment is high and risk of severe GVHD is low, regardless of donor type. For recipients of HLA-matched or HLA-1 antigen-mismatched sibling donor umbilical cord blood grafts (n = 62) as reported to the International Cord Blood Transplant Registry, the probabilities of engraftment and grade III-IV GVHD are 0.91 +/- 0.02 and 0.01 +/- 0.01, respectively. For recipients of HLA-matched or HLA 1-3 antigen-mismatched unrelated donor umbilical cord blood grafts (n = 85) transplanted at Duke University and University of Minnesota, the probabilities of engraftment and grade III-IV GVHD are 0.94 +/- 0.08 and 0.10 +/- 0.03, respectively. Notably, extensive chronic GVHD has yet to be reported in either series. The overall survival in recipients of unrelated donor umbilical cord blood grafts is 0.40 +/- 0.12 at 2 years after transplantation. Notably, no risk factor predicted higher risk of GVHD and only cell dose (P = 0.04) and possibly, degree of HLA mismatch (P = 0.06) impacted survival in multiple regression analysis. Although these data verify earlier reports that sufficient numbers of hematopoietic stem cells exist in umbilical cord blood for most recipients and that risks of acute and chronic GVHD are low despite increased HLA disparity, these data also provide us with a statistical analysis indicating which demographic, graft, and treatment factors possibly influence various outcomes after transplantation. PMID- 9358999 TI - Use of stem cells from mismatched related donors. AB - Mismatched haploidentical bone marrow transplantations from a related donor have been the topic of clinical and laboratory research for more than 20 years. During that time, new treatment strategies have been designed based on animal experiments, and, since our group introduced the megadose inoculum which combines T-cell-depleted bone marrow cells with a large number of granulocyte colony stimulating factor-mobilized peripheral blood stem cells and a more intensive conditioning regimen, have done much to overcome the problems of graft-versus host disease and graft rejection. As most patients have a full haplotype mismatched relative available, this technique means that a far greater number of patients with hematologic malignancies can be offered a T-cell-depleted transplantation as curative therapy. PMID- 9359000 TI - Purging marrow or peripheral blood stem cells for autografting. AB - Myeloablative therapy followed by autologous bone marrow or peripheral blood stem cell rescue is an alternative therapeutic option for patients who are not candidates for allogeneic transplantation due to the lack of an appropriate HLA matched donor or to advanced age. It allows therapy to be administered that is of comparable intensity to that used for allogeneic transplantation, and offers an alternative approach to achieving long-term disease-free survival. Although autologous transplantation obviates the risk of graft-versus-host disease, with its increased morbidity and mortality, it may also increase the rate of disease relapse due to the lack of an immune-mediated graft-versus-leukemia effect, as occurs in the allogeneic transplantation setting. The possibility that residual occult neoplastic cells, reinfused with the remission autografts following myeloablative therapy, can potentiate disease relapse has also been a concern in patients who undergo autologous bone marrow transplantation. This review discusses recent data concerning various strategies that are being used to "purge" autografts in patients undergoing autologous bone marrow transplantation in an effort to decrease the relapse rates and improve the disease-free and overall survival. PMID- 9359001 TI - Recent developments in the long-term preservation of red blood cells. AB - The first study to suggest the successful prolongation of the useful shelf life of red blood cells (RBCs) used a hypotonic additive solution containing glycerol. It was necessary to use twice the volume of this solution than the commercial additive solution per unit of packed RBCs. The final concentration of glycerol in the units was approximately 0.69% (75 mmol/L). A subsequent study demonstrated that the membranes of the exocytic microvesicles shed during storage had less of bands 3 and 4.1 than those in Adsol (Fenwal Laboratories, Deerfield, IL). Band 4.1 is important for strengthening the bonds between spectrin and actin in the cytoskeleton. In another study, glutamine or glutamine plus phosphate was used in a hypotonic additive solution, otherwise similar to the glycerol-containing additive. In the latter medium, phosphatidylethanolamine was less accessible to phospholipase action than in Adsol or when glutamine alone was added. Another group reported encouraging data regarding the action of L-carnitine when added to AS-3. Acylation of phosphatidylethanolamine mediated by the action of carnitine fatty acid transferase with acyl coenzyme A (acyl-CoA) occurred. Adenosine-5' triphosphate levels and red cell recovery were better in the test units. In the last paper reviewed, the authors demonstrated that oxidant damage of erythrocytes was less if the donors were given a mixture of antioxidants for 10 days prior to donating. PMID- 9359002 TI - Transfusion-associated iron overload. AB - Iron overload develops mainly via two mechanisms, by a defect in the regulation of iron absorption (hereditary hemochromatosis) or by parenteral route (chronic red cell transfusion for anemic patients without blood loss) especially in patients with different categories of refractory anemias, and in anemic patients with chronic infection, alcohol excess, and malignancies. The accurate assessment of body iron is indispensable for the correct diagnosis and for finding the optimal treatment schedule for each individual patient. Liver biopsy with quantitative iron determination and histochemistry is still the reference method for the assessment of body iron status for patients with iron overload. New noninvasive measurements (hepatic magnetic susceptibility, CT, and magnetic resonance imaging) are still investigational procedures. It is important to decrease the need for transfusion by judicious use of red cell concentrates, make more widespread use of erythrocytapheresis, determine the red blood cell phenotype of the patient before the onset of a regular transfusion regimen, treat concomitant hepatitis infections, consider splenectomy to diminish red blood cell requirements, and early on consider allogeneic bone marrow transplantation for thalassemic patients who have HLA-identical siblings. It is advisable to screen for the hereditary hemochromatosis gene before starting any kind or regular red blood cell transfusion therapy, and to avoid if possible, the risk of free radical release by transfusional iron overload during the physiologically hypercoagulable state of pregnancy and its effects on the highly proliferative tissues of the fetus. PMID- 9359003 TI - Transfusion-associated graft-versus-host disease. AB - Transfusion-associated graft-versus-host disease (TA-GVHD) is a devastating immunologic complication of blood transfusion. Patients at highest risk include premature infants and other patients who are immunosuppressed as a result of either congenital or acquired disease or because of the administration of immunosuppressive therapy. An additional high-risk group is immunocompetent patients who are heterozygous at a particular HLA locus and who receive blood from a donor who is homozygous at the same locus. The clinical syndrome consists of fever, skin rash, diarrhea, hepatic dysfunction, and bone marrow aplasia. The outcome is nearly always fatal, despite attempted treatments that have included the use of immunosuppressive agents. Hemorrhage and infection are the most common causes of death. Both humoral and cytotoxic mechanisms have been implicated in the pathophysiology of TA-GVHD. The complication of TA-GVHD can be prevented by the use of irradiated blood components. The use of ultraviolet B light-irradiated blood products and leukoreduction filters are also being investigated as potential preventive treatment modalities. PMID- 9359005 TI - Neutrophil antibodies. AB - Several advances have been made in understanding the polymorphisms of the neutrophil Fc-gamma-receptor IIIb (Fc gamma RIIIb). In one recent study, 21 individuals whose neutrophils lack Fc gamma RIIIb were found to be missing the entire Fc gamma RIIIB and Fc gamma RIIC genes. Another polymorphism of Fc gamma RIIIb, SH, has been characterized. New methods to determine the genotype of Fc gamma RIIIB for NA1, NA2, and SH using leukocyte genomic DNA have been described. A new monoclonal antibody to neutrophil-specific antigen NB1 was produced. Advances have been made in understanding alloimmunization in granulocyte transfusion recipients and the treatment of autoimmune neutropenia with granulocyte colony-stimulating factor (G-CSF). Granulocyte transfusion recipients were found to be alloimmunized both to neutrophil-specific and HLA antigens, suggesting that the transfusion of these patients with granulocytes matched only for HLA antigens will not be effective. A case report suggests that the beneficial effects of G-CSF on patients with autoimmune neutropenia is due in part to G-CSF's action of increasing plasma levels of soluble Fc gamma RIIIb. PMID- 9359004 TI - Transfusion in HIV disease. AB - Cytopenias often accompany infection with HIV. The pathophysiology responsible for the anemia, neutropenia, and thrombocytopenia in AIDS is complex and incompletely understood. Although HIV-infected patients with advanced disease were once routinely transfused for anemia accompanying zidovudine therapy, this practice has decreased as a result of decreased doses of zidovudine now being used and the more frequent use of HIV protease inhibitors. A more thorough understanding of the adverse effects of transfusion and the potential of immune modulation and facilitation of viral replication has no doubt led physicians to consider more thoroughly other factors before transfusing their patient. The results of trials examining the effect of leukodepletion on transfusion-related immune modulation are eagerly awaited. The availability of hematopoietic growth factors has also led to more effective treatment of the cytopenias without significant complications. PMID- 9359006 TI - Blood transfusion in disasters, war, and emergencies. AB - Historically, the needs of those wounded in war have led to many major advances in blood transfusion. The most important of these is probably the ability to draw blood in one location and transfuse it, at a later time, in a distant location. Another important lesson is the need for meticulous planning. Every hospital and blood center should have a disaster plan consisting of five components. What fluids to use, from where are they to be obtained, to what degree are they to be tested, how will they be transported to the disaster scene, and, once there, how will they be stored. Once drawn up, this plan must be regularly exercised and periodically revised. This will ensure rapid and efficient implementation when an emergency arises. Triage is vital in mass casualty situations, ensuring that scarce resources are used for those with the best chance of recovery. Although patients survive with low hemoglobin levels for considerable periods, speedy treatment of hypovolemia is imperative. When perflourochemicals and hemoglobin solutions are available for general clinical use, they will have a major role to play in disasters. Similarly, a simple method for the cryopreservation of red cells will allow stockpiles to be established. Unfortunately, none of these are presently available, although some are undergoing clinical trials. PMID- 9359007 TI - Transfusion-related bacterial sepsis. AB - Transfusion-related bacterial sepsis, although infrequent, is a serious and sometimes fatal transfusion complication. Several new studies confirm previous observations of the prevalence of contamination in red cell and platelet components. In addition, several recent reports have described the risk of bacterial contamination of hematopoietic progenitor preparations. Other studies have investigated potential interventions including development of alternative skin cleansing methods, alteration of whole blood storage time, cold storage of platelet suspensions, leukoreduction of red cells, development of rapid screening tests for bacteria, and a method for bacterial inactivation of platelet components. A generic approach to inactivation, such as those targeting bacterial nucleic acid, is particularly attractive because this method would not depend on the strain or source of contamination. PMID- 9359008 TI - Alloimmune refractoriness to platelet transfusions. AB - Patients who are transfused on multiple occasions with red cells or platelets may develop platelet-reactive alloantibodies and experience decreased clinical responsiveness to platelet transfusion. This situation, conventionally described as "refractoriness to platelet transfusions," is defined by an unsatisfactory low post-transfusion platelet count increment. If antibodies to HLAs are detected, improved clinical outcomes may result from transfusions of HLA-matched or donor recipient cross-matched platelets. Because refractoriness is an expected, frequently occurring phenomenon, prevention of HLA alloimmunization is an important management strategy. Prevention strategies include efforts to decrease the number of transfusions, filtration of cellular components to reduce the number of HLA-bearing leukocytes, or pretransfusion ultraviolet B irradiation of cellular components to decrease their immunogenicity. Other investigational approaches include reducing the expression of HLAs on transfused platelets, inducing a transient reticuloendothelial system blockade by infusions of specialized immunoglobulin products, or transfusing semisynthetic platelet substitutes (thromboerythrocytes, thrombospheres) or modified platelets (infusible platelet membranes, lyophilized platelets). PMID- 9359010 TI - Transfusion medicine. PMID- 9359009 TI - Hematopoietic stem cell transplantation. PMID- 9359011 TI - The evolving genetics of hepatitis C virus: separating the wheat from the chaff. PMID- 9359012 TI - Genetic heterogeneity of hepatitis C virus and its clinical implications. PMID- 9359014 TI - Inflammatory bowel disease and colon cancer: a review. AB - Extensive and long-standing colitis due to Crohn's disease and ulcerative colitis is associated with an increased incidence of colorectal cancer. Colonoscopy with biopsies for mucosal dysplasia can help stratify those patients who are at increased risk. However, the effectiveness of surveillance programs has been questioned. Newer molecular techniques may eventually lead to the development of more accurate screening tools, but at this time there is not enough evidence to support their widespread use. PMID- 9359013 TI - G protein-coupled receptor signaling: implications for the digestive system. AB - Extracellular signaling molecules regulate intracellular events by way of complex transduction assemblies composed of several proteins: receptor, G protein, effector, inactivating enzyme. Much is known about the structure and function of these transducer proteins. A signaling molecule initiates transduction by binding to the receptor which then prompts the G protein to undergo a reaction cycle. This cycle involves guanine nucleotide binding and hydrolysis, G protein subunit dissociation, and interactions with an effector (e.g. adenylyl cyclase, phospholipase C), as well as with inactivating molecules. The result is altered generation of intracellular second messengers, protein transcription, or another profound cellular response. This signal transduction system also contains multiple mechanisms for turning off the signal such as phosphorylating, internalizing, or downregulating receptors, uncoupling the receptor-G protein complex, or cell-surface peptidases, and precipitating conformational changes in transducer elements. These aspects of signal transduction are examined in two well studied systems, namely the beta 2-adrenergic and the substance P transducers. Both complexes are important physiological neuroregulators in the gut and elsewhere. Pathophysiological mechanisms involving aberrent signal transduction have been implicated in various diseases including major common illnesses such as heart failure and gastrointestinal disorders such as cholera, other infectious diarrheas, and colitis. PMID- 9359015 TI - Gastrointestinal motility considerations in patients with short-bowel syndrome. AB - Short-bowel syndrome results from large resections of the small intestine that result in the malabsorption of nutrients and fluids. Following intestinal resection both morphological and functional adaptations of the residual intestine occur. While we have witnessed progress in the understanding of morphological adaptation, little is known about the effects of gastrointestinal motility in short-bowel syndrome. This article reviews what is currently known about gastrointestinal motility in the context of short-bowel syndrome and the motility considerations that impact on clinical management. PMID- 9359016 TI - Abnormal gastric and small intestinal motor function in diabetes mellitus. AB - It is now well recognized that the prevalence of delayed gastric emptying in both insulin-dependent as well as noninsulin-dependent diabetes mellitus is high. Recently performed studies have shown that motor disorders of several parts of the upper gastrointestinal tract contribute to this delay in gastric emptying. Traditionally, disordered motility in diabetes mellitus has been attributed to irreversible autonomic nerve damage. However, recent observations indicate that hyperglycemia causes a reversible impairment of motility in various regions of the gastrointestinal tract. Upper gastrointestinal symptoms are highly prevalent in diabetes mellitus. These dyspeptic symptoms are not only induced by delayed gastric emptying, but altered visceroperception also plays a role in the genesis of dyspeptic symptoms. There is increasing evidence that impaired gastric emptying influences glycemia control, but the clinical consequences of these observations need further investigation. At present dyspeptic symptoms form the rationale for the treatment of delayed gastric emptying with prokinetic drugs. PMID- 9359017 TI - Molecular and genetic advances in gastrointestinal cancer: state of the art. AB - There have been major advances in the understanding of the molecular and genetic basis of gastrointestinal malignancies. The introduction of new genetic techniques has generated considerable insight into the understanding of familial cancer as is the case with hereditary nonpolyposis colon cancer and familial adenomatous polyposis syndrome and in the understanding of premalignant conditions such as the various dysplasias and adenomas. This new information will be of tremendous help in the early detection, the prognostication, and hopefully the therapy of premalignant and malignant gastrointestinal diseases. This article will attempt to review all these advances with added emphasis on the clinically relevant issues. PMID- 9359018 TI - Are right- and left-sided colon neoplasms distinct tumors? AB - Cancer of left and right colon has a differing prevalence at varying ages, in high- and low-incidence nations, as well as in men and in women. There also is a difference in clinical presentation, in prognosis, and possibly in genetic and environmental epidemiology. This review proposes that cancers of proximal and distal colon are different tumors because of their embryologic origin, genetic changes, and biologic identity. These factors are important in understanding the 'shift of tumors from more distal to more proximal sites in the colon' and in evaluating potential suggestions for instituting advances in diagnosis and prevention. PMID- 9359019 TI - Amiodarone hepatotoxicity. PMID- 9359020 TI - Genetic factors in osteoporosis. What are the implications for prevention and treatment? AB - Osteoporosis is a common disease that affects 1 in 3 women. Family and twin studies have demonstrated that there is a strong genetic component to this condition. Potential candidate genes examined for their regulatory effect on bone mass include those for collagen type I, estrogen and vitamin D receptors, and various cytokines and growth factors. To date, most work has focused on the vitamin D receptor (VDR) gene, and experience with this locus will probably act as a model for many future studies. There is increasing evidence, from population studies that have examined the relationship between VDR genotype and bone mineral density, of genetic heterogeneity and gene-environment interactions. Response to therapeutic agents may also be affected by an individual's underlying genotype, partly explaining the range of responses that are commonly observed in clinical practice. Knowledge of a person's genotype could, therefore, allow current therapies to be targeted to those most likely to benefit, with a possible reduction in adverse effects. Largescale genomic studies of osteoporosis may also identify novel genes, and this may lead to both a better understanding of disease pathophysiology and to the discovery of potential targets for drug development. PMID- 9359021 TI - Drug therapy for obesity in the elderly. AB - The prevalence of obesity is increasing rapidly in the US and other developed countries. Even though the percentage of older individuals is increasing worldwide, obesity has only recently become a recognised problem in this population. Obesity occurs when energy intake chronically exceeds energy expenditure. Moreover, advancing age is associated with an inability to couple energy intake with energy expenditure. Obesity contributes to many adverse health outcomes, including non-insulin-dependent (type II) diabetes mellitus, as well as to an increase in both cardiovascular and all-cause mortality. Only recently has the medical community begun to accept obesity as a disease with a multifactorial pathogenesis that requires systematic lifestyle changes and pharmacological treatment. Several groups of drugs are available for the pharmacotherapy of obesity; anorectic medications (e.g. fenfluramine, dexfenfluramine); substances affecting energy expenditure and body composition [e.g. chromium (chromium picolinate), ephedrine, anabolic steroids, beta 3-adrenoceptor agonists]; and drugs affecting the absorption of nutrients (e.g. orlistat). To date, few drugs have produced and sustained a significant bodyweight loss. However, some drugs induce a significant short term reduction in bodyweight compared with placebo. Moreover, there is a paucity of information regarding the effectiveness of these drugs in the treatment of obesity in the elderly. Furthermore, it is even debated whether obesity should be treated with drug intervention in the elderly. Clinicians prescribing medications for obesity treatment in the elderly need to carefully consider the benefit: risk ratio, given the high prevalence of polypharmacy in elderly patients. Furthermore, physiological changes that occur with aging may affect the pharmacokinetics of administered drugs and need to be taken into consideration. PMID- 9359022 TI - Systemic adverse effects of topical ophthalmic agents. Implications for older patients. AB - Topical ophthalmic medications are widely prescribed by growing numbers of eye care professionals. Increasingly, these agents are being prescribed by optometrists and ophthalmic-trained nurses in addition to ophthalmologists and general practitioners. As the number and variety of topical agents on the market rises, and as the number of clinicians involved in prescribing those agents increases; the risk of systemic adverse effects will also increase. Thus, professionals involved in the care of these patients must be aware of the risks associated with these drugs in order to minimise the likelihood of complications. Moreover, inadequate training may result in the clinician failing to associate a topical medication with a systemic condition, allowing an adverse effect to pass unrecognised. It is therefore in the interest of the ophthalmic and pharmaceutical communities to improve awareness of the potential dangers intrinsic in the use of topical eye medications. It is the elderly population who are at greatest risk of experiencing systemic adverse effects of topical agents. Chronic ophthalmic diseases, and hence long term ophthalmic drop treatments, are more prevalent among older people. Such individuals are also likely to have other medical conditions (e.g. cardiac, respiratory or neurological disease) that may be induced or exacerbated by topical ophthalmic agents. Moreover, polypharmacy is common in elderly people, and this is associated with an increased risk of drug interactions. PMID- 9359023 TI - Classification, diagnosis and treatment of vascular dementia. AB - Vascular dementia (VAD) is considered to be the second most common cause of dementia in Europe and the US. In Asia and many developing countries, it is more common than dementia of the Alzheimer's type (DAT). VAD is the most preventable form of dementia associated with later life. The pathogenesis of VAD is multifactorial, and it represents a heterogeneous, not a homogeneous, clinical entity. Classification of VAD by pathogenesis is important for its prevention and treatment. Control of the risk factors for VAD reduces its incidence and stabilises or improves cognitive performance following stroke. Proper diagnostic evaluation of VAD requires: (i) a well defined quantitative assessment of the cognitive deficits present; (ii) assessment of risk factors for stroke; (iii) identification of cerebral vascular lesions by history, neurological examination and neuroimaging; (iv) exclusion of other causes of dementia; (v) establishment of a positive diagnosis of possible, probable or definite VAD versus DAT or mixed VAD/DAT; and (vi) identification of the temporal relationship between cognitive deficits and cerebral vascular lesions. VAD can be subdivided into 8 major types, as follows: (i) multi-infarct dementia secondary to large cerebral emboli [type 1]; (ii) strategically placed infarctions causing dementia [type 2]; (iii) multiple subcortical lacunar lesions secondary to atherosclerosis or degenerative arteriolar changes [type 3]; (iv) Binswanger's disease (arteriosclerotic subcortical leukoencephalopathy) [type 4]; (v) mixtures of types 1, 2 and 3 [type 5]; (vi) haemorrhagic lesions causing dementia [type 6]; (vii) subcortical dementia secondary to hereditary factors (type 7); and (viii) mixtures of DAT and VAD (type 8). Treatment is dictated by the pathogenetic subtype of VAD that is present. PMID- 9359026 TI - Samarium 153Sm lexidronam. AB - Samarium 153Sm lexidronam is a medium energy beta-emitting radioisotope chelated to the tetraphosphate, ethylenediaminetetramethylenephosphonic acid (EDTMP). 153Sm has a physical half-life of 1.9 days. Samarium 153Sm lexidronam concentrates in bone tissue and has a metastatic lesion: normal bone ratio of 4. Clearance of nonskeletal radioactivity after administration of samarium 153Sm lexidronam is rapid, and is almost complete from the blood and urine within 1 and 6 hours, respectively. Some degree of pain relief was achieved in 72% of patients with bone metastases after a single dose of samarium 153Sm lexidronam 1 mCi/kg (37 MBq/kg) compared with 44% of placebo recipients (p < 0.025). Relief was generally attained within 2 weeks and lasted for a median of 8 to 15 weeks. After recurrence of pain, a second dose of samarium 153Sm lexidronam may produce further pain relief in some patients. The dose-limiting toxicity of samarium 153Sm lexidronam is reversible myelosuppression. PMID- 9359027 TI - Cyclins: relevance of subcellular localization in cell cycle control. PMID- 9359025 TI - How do drugs relieve neurogenic pain? AB - Neurogenic pain is experienced by about 1% of the population. The efficacy of drug treatment for this condition has been poorly evaluated, and only recently have certain treatments been shown to have significant analgesic effects. Monotherapy with topical agents such as capsaicin is not usually sufficient. Oral agents that have proven effective in treating neurogenic pain states include tricyclic antidepressants, selective serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitors and anticonvulsants. Local anaesthetics, administered intravenously, have been reported to relieve pain in selected patients, but data from controlled trials are sparse. Multiple mechanisms contribute to the generation of neurogenic pain. In the future, drug treatment for neurogenic pain is likely to target these mechanisms. Recent studies have shown that N-methyl-D aspartate (NMDA) receptor antagonists, adenosine receptor agonists and nitric oxide synthase inhibitors may become useful in the treatment of neurogenic pain. PMID- 9359028 TI - PP60c-src expression in osteoclasts from osteopetrotic children and in giant tumor cells. AB - Malignant infantile osteopetrosis is a severe congenital disease characterized by impaired osteoclast activity. Among the multiple factors that influence bone resorption, the c-src protooncogene product pp60c-src plays an essential role, since mice which lack pp60c-src develop osteopetrosis. To gain insight into the possible role of pp60c-csrc in the pathogenesis of infantile osteopetrosis, we examined the osteoclasts of three children displaying the typical features of the disease, aged respectively one, four and seven months. pp60c-csrc expression and localization, together with the expression of a 80/85-kilodalton pp60c-src substrate, cortactin, were examined by immunoelectron microscopy. Osteoclasts from two giant cell tumors were used as controls. Bone and tumor samples were fixed in 4% paraformaldehyde, included in LR-White resin at -30 degrees C and the sections processed with mAb 327 or mAb anti p80/85 by an immunogold technique. pp60c-src was expressed in the cytoplasm, in nuclear membranes and in nuclei of the osteoclasts of the three osteopetrotic children. The subcellular localization of the kinase was not different from the localization in giant tumor cells. In both cases cortactin was abundant. In conclusion, in three children with malignant osteopetrosis, pp60c-src expression in osteoclasts does not appear to be involved in the pathogenesis of the disease. The presence of this protein, however, does not necessarily reflect normal c-src tyrosine kinase activity, nor normal c-src-dependent intracellular signaling pathways. Moreover the presence of the protein in nuclear membranes, and especially around nuclear pores supports the hypothesis that in osteoclasts, c-src may participate in the regulation of RNA processing. PMID- 9359029 TI - MC22-33F monoclonal antibody shows unmasked polar head groups of choline containing phospholipids in cartilage and bone. AB - The importance of the role lipids may have in biological calcification, and the paucity and poor specificity of the methods available for studying their ultrastructural localization, call for further investigations involving new techniques. The monoclonal antibody MC22-33F displays a specific reaction with phosphatidylcholine, sphingomyelin and dimethylphosphatidylethanolamine, as confirmed by its reaction with synthetic lyposomes of pure phosphatidylcholine. For this reason, it was used to demonstrate the presence and ultrastructural localization of choline-containing phospholipids in calcifying cartilage and bone. The MC22-33F MAb immunoreaction was found in the cytoplasm of maturing and hypertrophic and, to a lesser extent, proliferating and degenerating chondrocytes; it was strongly positive in matrix vesicles, at the periphery of calcification nodules, and at the periphery of calcified matrix. In bone, the immunoreaction was especially strong along the peripheral membrane of the osteoblasts, in the interfibrillary spaces of the osteoid border, in matrix vesicles and in the peripheral zone of the calcification nodules. The central zone of the nodules, the fully calcified matrix and most of the intracellular membranes were negative in both cartilage and bone. These results confirm the presence of choline-containing phospholipids in early areas of calcification, including matrix vesicles. They also show that the intracellular membrane phospholipids are not accessible to the antibody, possibly because they are masked by other substances. These are not proteoglycans, because their enzymatic digestion does not increase or modify the reactivity of MC22-33F MAb. In spite of this limitation, MC22-33F MAb offers considerable advantages in the study of the calcification process, because the positive immunoreaction of the calcifying bone matrix, and the negativity of calcified matrix, confirm without doubt that phospholipids have an active role in biological calcification. PMID- 9359031 TI - Dating of fibrotic lesions by the Picrosirius-polarization method. An application using the lesions of Lechiguana (bovine focal proliferative fibrogranulomatous panniculitis). AB - This work was designed to verify if a simple quantitative procedure to estimate the state of collagen aggregation is useful in describing the natural history of a fibrous process. For this purpose sixteen cases of Lechiguana lesion were used. Histochemical evaluation of the collagen content and its state of aggregation was done by the Picrosirius-polarization method. Morphometric studies were done by means of a point-counting procedure, which allowed the determination of the areal fraction of thin and thick collagenous fibers within Lechiguana lesions collected at different times of clinical evolution (14 days through 8 months). Early lesions are characterized by thin collagenous fibers. This population of slender fibers decreases later on, when thick fibers become more prevalent. Curve fitting procedures were employed using the state of collagen aggregation as the dependent variable and time as the independent variable. The best fitting was obtained by linear and exponential functions. Statistical analysis indicates that the quantitative study of the degree of collagen organization allows an adequate determination of the time course of Lechiguana lesions. We concluded that simple determinations of collagen aggregation provide numerical data that may be useful to build mathematical models relating time of evolution of the disease to fibrosis. PMID- 9359032 TI - Detection of apoptotic cells by annexin V labeling at electron microscopy. AB - Annexin V is a Ca(++)-binding protein which is widely used as a marker for apoptotic cells, as it binds to the phosphatidylserine residues exposed at the surface of apoptotic cells. In this paper, we describe a method for the immunogold-labeling of biotin-conjugated Annexin V, to detect apoptotic thymocytes at electron microscopy. Etoposide-treated thymocytes were reacted in tissue culture medium with biotin-conjugated Annexin V, fixed with glutaraldehyde, and processed for resin embedding; thin sections were incubated with antibiotin antibodies coupled with colloidal gold. Cytometric estimates of the apoptotic index were also performed by evaluating either the DNA content after propidium iodide staining or the light-scattering values, as well as the positivity for fluorescein isothiocyanate-conjugated anti-biotin antibodies. At electron microscopy, gold-labeling of Annexin V was located on the plasma membrane only of apoptotic thymocytes and on cytoplasmic debris, likely resulting from the typical apoptotic blebbing. Unlabeled thymocytes always showed normal, non-apoptotic nuclear morphology. The application of Annexin V labeling at electron microscopy will allow a more refined description of the morphological events occurring during apoptosis. PMID- 9359030 TI - Potential role of streptozotocin in enhancing ossification of the posterior longitudinal ligament of the cervical spine in the hereditary spinal hyperostotic mouse (twy/twy). AB - We investigated the effects of streptozotocin-induced diabetes mellitus on ossification of the posterior longitudinal ligament (OPLL) in the murine cervical spine. A genetically-bound spinal hyperostotic mouse, the tip-toe walking Yoshimura (twy) mouse, was used in these experiments. Histological examination showed that streptozotocin enhanced membranous and enchondral ossification of the posterior longitudinal ligament of the cervical spine, particularly in the area of the ligamentous enthesis. It also increased the number of alkaline phosphatase positive osteoblast-like mesenchymal cells particularly around the enthesis, while the number of such cells was less in control twy mice and ICR mice treated with streptozotocin. The area of OPLL subsequently increased in size in streptozotocin-treated twy mice. We suggest that streptozotocin-induced diabetes enhances OPLL in the genetically-bound spinal hyperostotic mouse (twy/twy). PMID- 9359033 TI - Immunocytochemical identification of cells containing insulin, glucagon, somatostatin, and pancreatic polypeptide in islets of Langerhans of the wombat pancreas with electron microscopy. AB - The endocrine pancreas of the southern hairy-nosed wombat (Lasiorhinus latifrons) was investigated by means of electron-microscope immunocytochemistry using the protein A--gold technique on London resin (LR) white-embedded tissue. The primary antibodies used were raised against insulin, glucagon, somatostatin and pancreatic polypeptide. The morphology of the secretory granules differed in the four cell types. The insulin cells are pleiomorphic, and the secretory granules composed of an electron-dense core surrounded by an electron-lucent halo. The glucagon cells possess granules of varying density. Somatostatin cells have large, less dense secretory granules. The pancreatic polypeptide cells show small dense secretory granules. It is essential that it be corroborated by immunocytochemical data at the light or preferably electron-microscopical level for the definite identification of endocrine cell types. Recent developments in immuno-electron-microscopic techniques have contributed to a better knowledge of cells responsible for the secretion of a wide variety of hormones, as in this study. PMID- 9359024 TI - Immunodeficiency of aging. AB - Aging is associated with declines in multiple areas of immune function, but to date no single mechanism has emerged as being responsible for all the observed changes. Many changes occur at different rates within individuals as well as between individuals. With advancing age there is a concomitant increase in the incidence of many infections and cancers. It is being increasingly acknowledged that autoimmune processes play a proinflammatory role in the development of many pathological conditions, such as atherosclerosis. However, direct causal relationships between specific changes in immunity and the occurrence of specific diseases are rare. There is accumulating epidemiological, in vivo and in vitro evidence to support many such direct relationships in both animals and humans. It is likely that the mechanisms underlying age-related changes in immunity are multifactorial, with both genetic and environmental factors playing a significant role. Despite the current lack of unifying theories, much active and exciting work is proceeding in the area of immune stimulation. Studies describing age related changes in immunity, as well as the testing of interventions to reverse these changes, will continue to fill the gaps in our knowledge, leading to a more comprehensive understanding of immunosenescence. PMID- 9359034 TI - Effects of cocaine treatment on the nervous system of planaria (Dugesia gonocephala s. l.). Histochemical and ultrastructural observations. AB - Acute high dose treatment with cocaine in planaria has been shown to produce hyperkinesia followed by immobilization, thus suggesting progressive neuronal dopamine (DA) depletion. On the contrary, treatment with low doses of cocaine inhibits motor activity in planaria, without producing hyperkinesias. Here we investigated the morpho-functional changes of the DA presynaptic terminals following cocaine treatment in planaria (acute high dose and chronic low dose). Neuronal DA content was determined by means of histochemical methods, and nerve cell ultrastructure was examined by electron microscopy. The effects of cocaine were compared to those of L-dopa, reserpine (used as positive and negative controls, respectively) and normal untreated specimens. Presynaptic vesicles and DA content were significantly reduced by chronic low-dose cocaine treatment. These effects were even more robust when the drug was acutely administered at high dose. Thus, depletion of DA vesicles is produced by cocaine in planaria, as well as in mammals. The behavioral effects of chronic low-dose treatment with cocaine, however, suggest that the drug acts not only as a DA reuptake blocker, but also as a direct agonist on presynaptic DA receptors. Acute high-dose administration of cocaine also produced signs of neuronal suffering, thus providing evidence for a direct neurotoxic effect of the drug. PMID- 9359035 TI - Double labeling on the two faces of the same section with lectin-gold and silver enhancement. Ultrastructural detection of the terminal disaccharides sialic acid beta-galactose and sialic acid-alpha-N-acetylgalactosamine. PMID- 9359036 TI - Mutations in Hirschsprung disease: when does a mutation contribute to the phenotype. AB - Hirschsprung disease is a congenital disorder clinically characterized by the absence of colonic ganglia and genetically by extensive heterogeneity. Genes involved include RET, GDNF, EDNRB and EDN3. Mutations of these genes may give dominant, recessive, or polygenic patterns of inheritance. In particular in the case of missense mutations, it is therefore far from easy to assess whether a given mutation will contribute to the phenotype. We discuss criteria for such an assessment and pay special attention to functional assays. The interpretation of mutations as contributing to a disease phenotype or as merely representing a rare polymorphism has direct clinical consequences. Hirschsprung disease with major and modifying sequence variants in a variety of genes might well serve as a model for the many complex disorders for which the search for genes involved has only just been initiated. PMID- 9359038 TI - Two new mutations in the glucose-6-phosphatase gene cause glycogen storage disease in Hungarian patients. AB - Glycogen storage disease type 1a (von Gierke disease, GSD-1A) is caused by the deficiency of microsomal glucose-6-phosphatase (G6Pase) activity which catalyzes the final common step of glycogenolysis and gluconeogenesis. The cloning of the G6Pase cDNA and characterization of the human G6Pase gene enabled the identification of the mutations causing GSD-1a. This, in turn, allows the development of non-invasive DNA-based diagnosis that provides reliable carrier testing and prenatal diagnosis. Here we report on two new mutations E110Q and D38V causing GSD-1a in two Hungarian patients. The analyses of these mutations by site-directed mutagenesis followed by transient expression assays demonstrated that E110Q retains 17% of G6Pase enzymatic activity while the D38V abolishes the enzymatic activity. The patient with the E110Q has G222R as his other mutation. G222R was also shown to preserve about 4% of the G6Pase enzymatic activity. Nevertheless, the patient presented with the classical severe symptomatology of the GSD-1a. PMID- 9359037 TI - A number of schizencephaly patients including 2 brothers are heterozygous for germline mutations in the homeobox gene EMX2. AB - We report here that some patients affected by schizencephaly are heterozygous for mutations in EMX2, a homeobox gene implicated in the patterning of the developing forebrain. Schizencephaly is a very rare human congenital disorder characterized by a full-thickness cleft within the cerebral hemispheres. Large portions of these may be absent and replaced by cerebrospinal fluid. We previously reported the presence of EMX2 mutations in 7 out of 8 sporadic cases of schizencephaly. We now extend this analysis to 10 additional patients, including 2 brothers. Six patients were found to be heterozygous for de novo mutations in EMX2. In particular, the 2 brothers show the same mutation affecting the splicing of the first intron, while this mutation is absent in their parents and in the 2 unaffected siblings. PMID- 9359039 TI - Relationship between mutation genotype and biochemical phenotype in a heterogeneous Spanish phenylketonuria population. AB - Genotyping of the phenylalanine hydroxylase (PAH) gene offers a new tool for characterizing patients with phenylketonuria (PKU), refining the diagnosis and aiding in the prediction of the clinical outcome and in the implementation of a more adequate treatment. The primary goal of this work was the detailed study of the different allele combinations and the metabolic phenotypes in Spanish PKU patients in order to understand better the clinical heterogeneity of PAH deficiency in our population. The results show that the disease phenotype is a consequence of a combination of mutations at the PAH locus and this observation is valid throughout the spectrum of clinical and biochemical varieties found in Spanish PKU patients. A stronger correlation was found between the predicted residual activity, when known from previous in vitro studies of the mutant proteins, and the Phe tolerance than between the predicted residual activity and the inverse of Phe levels at diagnosis. The observed genotype-phenotype correlations and the available data on the in vitro residual activity of the mutant proteins has enabled the estimation of the severity of most of the mutations found in Spain. This study includes relevant data for clinicians and pediatricians adding to the present knowledge which relates allelic PAH genotypes to biological phenotypes. PMID- 9359040 TI - Estimating recessive disease allele frequency based on genetic maps. AB - For a recessive disease whose gene has been localized on the human gene map, a new method is described for estimating the population frequency of the disease allele. The method focuses on affected individuals whose parents are first cousins, where parents and grandparents are genotyped for highly polymorphic markers at the disease gene. The primary statistic is the proportion of such probands who are autozygous (homozygous due to identity by descent of the two disease alleles), where this proportion is a function of the disease allele frequency. Our map-based method is compared to Dahlberg's method of estimating recessive disease allele frequencies, which is based on the proportion of affected individuals whose parents are first cousins; this proportion is also a function of the disease allele frequency. For small to moderate sample sizes and traits that are not too common, our new method is more efficient (for some parameter values dramatically more efficient) than Dahlberg's method. PMID- 9359041 TI - Refined mapping of the Cohen syndrome gene by linkage disequilibrium. AB - The Cohen syndrome is a rare autosomal recessively inherited disorder. Contrary to many case reports published elsewhere, the phenotype is uniform in Finland including nonprogressive mental and motor retardation, typical dysmorphic features, granulocytopenia and marked ophthalmological changes. By linkage analysis in five Finnish multiplex nuclear families, the COH1 locus for the Cohen syndrome was recently assigned to a 10-cM region between loci D8S270 and D8S521 on the long arm of chromosome 8. Here we present results of linkage disequilibrium and haplotype analysis in an extended panel of 16 Finnish COH1 families using new markers localized in the COH1 region. By inferring historical recombinations in conserved haplotypes the COH1 gene was assigned in the region of marker loci D8S1808, D8S1762 and D8S546. Calculations of genetic distances based on linkage disequilibrium suggest that the most likely localization of COH1 is in the immediate vicinity of marker locus D8S1762. Haplotype analysis suggests the occurrence of one main COH1 mutation and possibly one or two rare ones in Finland. This information will be useful in the positional cloning of the gene. PMID- 9359042 TI - Refined subchromosomal location of 21 expressed sequence tags from unknown genes at region 11p15. AB - Twenty-one expressed sequence tags (ESTs) belonging to unidentified transcripts were mapped to 9 intervals at 11p 15.5-p15.3. Thirteen were mapped to a region of an estimated size of 7 Mb with the help of a YAC contig. The remaining eight were mapped outside this region, and the centromeric or telomeric position of the ESTs relative to the YAC contig was defined with the help of a cellular hybrid containing a derivative chromosome 11 with a break-point within the region covered by the contig. The average size of the intervals, where the ESTs were mapped, was estimated to be 0.3-0.4 Mb. The refinement of the chromosomal location of these ESTs could be of great help in the identification of potential candidate genes for disease locus mapping at 11p15. PMID- 9359044 TI - Refined localisation of the genes for nebulin and titin on chromosome 2q allows the assignment of nebulin as a candidate gene for autosomal recessive nemaline myopathy. AB - A locus for autosomal recessive nemaline myopathy (NEM2) has been assigned by linkage analysis to a 13-cM region between the markers D2S150 and D2S142 on 2q21.2-q22. The genes for the giant muscle proteins nebulin and titin have previously been assigned by FISH to 2q24.1-q24.2 and 2q31, respectively. By using radiation hybrid mapping, we have reassigned the nebulin gene close to the microsatellite marker D2S2236 on 2q22 and the titin gene to the vicinity of the markers D2S384 and D2S364 on 2q24.3. The genomic orientation of the nebulin gene was determined as 5'-3' and of TTN as 3'-5' from the centromere. We conclude that the nebulin gene resides within the candidate region for NEM2 on the long arm of chromosome 2, while the titin gene is located outside this region. PMID- 9359043 TI - Three novel point mutations in the keratinocyte transglutaminase (TGK) gene in lamellar ichthyosis: significance for mutant transcript level, TGK immunodetection and activity. AB - We have investigated 8 patients from 7 unrelated families with lamellar ichthyosis (LI) for defects in the keratinocyte transglutaminase (TGK) gene. We have characterized three novel homozygous mutations and a previously reported splice acceptor site mutation. One patient showed a C-to-T change in the binding site for the transcription factor Sp1 within the promoter region. Another patient had a Gly 143-to-Glu mutation in exon 3 and a third patient, affected with a particular form of LI sparing the four limbs, demonstrated a Val382-to-Met mutation within exon 7. These three patients exhibited drastically reduced transglutaminase activity and an absence of detectable TGK polypeptide, as assessed by immunofluorescence and immunoblotting. Northern blot analysis showed that the Sp1 site mutation was associated with profound reduction of TGK transcript levels whereas normal transcript levels were observed for the two missense mutations. We hypothesize that the Sp1 site mutation impairs transcription of the TGK gene, whereas the two missense mutations induce structural changes leading to protein instability. Linkage to TGK was excluded in another family and no evidence for TGK defect was found in 3 other patients. These results further support the involvement of TGK in some patients with LI. They identify a TGK mutation as a cause for non-generalized LI and further delineate the molecular mechanisms underlying TGK deficiency in LI. PMID- 9359045 TI - Lack of hemizygosity for the insulin-like growth factor I receptor gene in a quantitative study of 33 Silver Russell syndrome probands and their families. AB - Previous studies have shown that individuals with a deletion of 15q26.1-->qter, which includes the insulin-like growth factor I receptor (IGFIR) gene, may exhibit phenotypic characteristics similar to those individuals with Silver Russell Syndrome (SRS). Thirty-three SRS probands, with normal karyotypes, and their parents were investigated for the presence of both copies of IGFIR by gene dosage analysis of Southern blot hybridisation. All 33 SRS probands have both copies of IGFIR. Tetranucleotide repeat marker analysis for three locations on 15q also ruled out other deletions in these regions for those markers that were informative. Two important functional regions of IGFIR were also investigated for DNA mutations, using single-stranded conformational polymorphism analysis. No mutations were found in the cysteine-rich region involved in ligand binding (exon 3) or the ATP binding region (exon 16) which could contribute to the SRS phenotype. However, a silent mutation in the third position of one of the codons in the ATP region (3174G-->A, 1013 Glu-->Glu) was found. PMID- 9359046 TI - BOR and BO syndromes are allelic defects of EYA1. AB - Branchio-oto-renal (BOR) syndrome is an autosomal dominant disease characterized by varying combinations of branchial, otic and renal anomalies. By positional cloning, a candidate gene, EYA1, homologous to the drosophila eyes absent gene, has recently been identified at 8q13.3 and shown to underlie this syndrome. The name branchio-oto (BO) syndrome has been used to describe a similar combination of branchial and otic anomalies, without the association of renal anomalies. Whether BOR and BO syndromes involve the same gene was unknown. To address this question, we analyzed two large independent families for which each of the 8 affected members present exclusively with BO syndrome. In both families, linkage analysis mapped the causative gene to the same chromosomal region as EYA1. A search for mutations in 9 of the EYA1 coding exons identified a 2-bp insertion segregating in one family and an 8-bp deletion segregating in the other. These results demonstrate that EYA1 also underlies BO syndrome, and that BOR and BO syndromes are allelic defects of this gene. PMID- 9359047 TI - Endothelin-3 gene mutations in isolated and syndromic Hirschsprung disease. AB - Hirschsprung disease (HSCR, aganglionic megacolon) is a frequent congenital malformation regarded as a multigenic neurocristopathy. Four susceptibility genes have recently been identified in HSCR, namely the RET proto-oncogene, the glial cell line-derived neurotrophic factor (GDNF), the endothelin B receptor (EDNRB) and the endothelin-3 genes (EDN3). Homozygosity for EDN3 mutations has been previously shown to cause the Shah-Waardenburg syndrome, a combination of HSCR with features of the Waardenburg syndrome. Here, we report on heterozygous EDN3 missense mutations in isolatec HSCR. The present data give further support to the role of the endothelin signaling pathway in the development of neural crest derived enteric neurons. They also suggest the possibility that either recessive or weakly penetrant dominant alleles could occur at the EDN3 locus, depending on the nature of the mutation. PMID- 9359048 TI - Factors increasing the risk of allograft vascular disease in heart transplant recipients. AB - Allograft vascular disease is the major cause of late cardiac graft failure. A multifactorial etiopathogenesis is supposed. Our study investigated factors associated with allograft vascular disease occurrence. After stratifying our series on the basis of potential risk factors, we calculated allograft vascular disease incidence rate in 267 grafts from 258 patients who underwent transplant between November 1985 and August 1996. Chi-square test was used for the identification of univariate risk factors to be included in a multivariate model. Multivariate analysis was based on a Poisson model. Seventy of the 267 grafts (26.2%) were diagnosed with allograft vascular disease. Heart disease other than idiopathic dilated cardiomyopathy, donor's age, number of mismatches for HLA-B = 2, presence of systo-diastolic hypertension, number of acute rejection positive endomyocardial biopsies > or = 7 and the association of human Cytomegalovirus and hepatitis C virus infections proved to be univariate risk factors, and were included in the Poisson multivariate model. The association of Cytomegalovirus and hepatitis C infections multiplied allograft vascular disease incidence rate by 3.9, systo-diastolic hypertension by 2.2, occurrence of 2 HLA-B mismatches by 2, a high number (> or = 7) of acute rejection positive-endomyocardial biopsies by 1.8, and heart disease other than idiopathic dilated cardiomyopathy by 1.8. The association of human Cytomegalovirus and hepatitis C virus infections, of HLA B mismatches, of acute rejection-positive endomyocardial biopsies, as well as post-transplantation hypertension and native heart disease other than idiopathic dilated cardiomyopathy, proved to be positively associated with an increased risk of allograft vascular disease. Given the concordance of our data with those of numerous prior series, we are going to adopt a special surveillance angiographic protocol for patients with these factors. PMID- 9359049 TI - Ultra-early risk stratification after myocardial infarction via pharmacological stress echocardiography: the relative value of resting function, viability and myocardial ischemia. AB - BACKGROUND: Echocardiographically recognized resting function, myocardial viability (by low-dose dobutamine) and stress-induced ischemia (by high-dose dipyridamole) are potent predictors of subsequent events, but their relative value in patients evaluated very early after acute myocardial infarction remains to be established. AIM OF THE STUDY: To assess the feasibility and usefulness of an integrated approach with resting and pharmacological stress echo for risk stratification in patients evaluated very early after myocardial infarction. METHODS: Sixty acute myocardial infarction patients without contraindications to stress testing, and who were being given thrombolytic therapy, underwent resting echo (16-segment model of left ventricle, each segment scored from 1 = normo hyperkinetic to 4 = dyskinetic), high-dose dipyridamole (up to 0.84 mg/kg over 10') and low-dose dobutamine (up to 10 mcg/kg/min) echo on the third-fourth day after drug withdrawal. The response was "ischemic" with a dipyridamole-induced increase in the regional score > 1 in segments with a resting score < 3, and "viable" with a dobutamine-induced decrease in the regional score > 1 in segments with resting score > 1. All patients underwent coronary angiography on the tenth twelfth day after the acute event, and all of them were followed up for 15 +/- 10.04 months. RESULTS: Ischemia elicited by dipyridamole appeared in 29 patients (48%) and dobutamine-induced viability was observed in 28 (47%). Ischemic events occurred in 26 patients (43.4%), five of which during the early in-hospital period. There were three deaths (5%), one re-AMI (1.7%), 7 Canadian Class III-IV angina (12%) and 15 (25%) early revascularization procedures undertaken independently of stress echo results. Events occurred in 21 patients (72%) with dipyridamole-induced ischemia and in 5 (16%) without it (p < 0.001). Likewise, events occurred in 13 patients (46.4%) with dobutamine-induced inotropic recovery and in 13 (40.6%) without it (p = ns). Event-free survival occurred in 64% of dipyridamole-positive patients, as opposed to 90% of dipyridamole-negative patients (p = 0.025). Dipyridamole echocardiographic test sensitivity and specificity for events were 81 and 74%, respectively. Sensitivity and specificity for events of dobutamine viability were 46 and 55%, respectively. In a multivariate logistic analysis, dipyridamole-induced myocardial ischemia was the strongest predictor of subsequent events (p = 0.01). According to Cox analysis, dipyridamole positivity had a relative risk estimate of 4. CONCLUSIONS: Pharmacological stress echo is feasible even very early after acute myocardial infarction via a useful approach based on low-dose dobutamine to assess myocardial viability, and high-dose dipyridamole to assess ischemia. For risk stratification purposes, stress-induced myocardial ischemia outperforms resting function and myocardial viability, and it is independent of angiographic data. Revascularization procedures do not seem to be effective when only viability is present. PMID- 9359050 TI - Risk stratification after acute myocardial infarction: "rich lion" or "poor sheep" approach? PMID- 9359051 TI - [A longitudinal study on left atrial thrombosis in patients with non-rheumatic atrial fibrillation treated with anticoagulants]. AB - BACKGROUND: Although atrial thrombosis is common in patients with non-rheumatic atrial fibrillation (NRAF) (6-27%), there are no studies about the effect that anticoagulant or antiplatelet drugs have on it. AIM OF THE STUDY: We have investigated the role of anticoagulant therapy, followed by family physicians, on left atrial thrombosis detected via transesophageal echocardiography (TEE) in patients with NRAF. METHODS: Sixty patients enrolled in the TASAF (Trieste Area Study on non-rheumatic Atrial Fibrillation) (60% males, mean age 72 +/- 7 years, 17% with lone atrial fibrillation, duration of arrhythmia 111 +/- 79 months), in whom we found a left atrial and/or left atrial appendage thrombus via TEE, were anticoagulated. The thrombus was in the left atrial appendage in 57 patients and in the left atrium in the others. In 28 of them it was mobile and in 50, the left atrial appendage flow was low or absent. Lastly, in 46 patients we found spontaneous echocontrast and a bilobate appendage was present in 8 of them. At follow-up, we repeated the TEE to evaluate the effect of the therapy on the atrial thrombosis. RESULTS: Only 53 patients received anticoagulant therapy (45 correctly), while 7 were treated with antiplatelet drugs by their family physicians. After a mean follow-up of 16 months, all patients underwent a repeat TEE and the thrombus was no longer evident in 35 cases (58%). The thrombosis disappeared in 26 (58%) of the 45 patients who correctly took anticoagulant therapy and in 5 of the 8 who were not adequately anticoagulated. Moreover, we didn't observe the thrombus in 4 of the 7 patients who were treated with antiplatelet drugs. There was no significant statistical difference between the benefits of anticoagulants and antiplatelet therapy in dissolving left atrial thrombosis. During follow-up, we recorded only one embolic event in the retina. None of the patients on antiplatelet drugs complained of any side effects, whereas in 5 of the 53 on anticoagulant therapy, we recorded one fatal intracranial hemorrhage, one gastric hemorrhage and three minor complications. CONCLUSIONS: Left atrial thrombosis in NRAF disappears in anticoagulated patients in an high percentage of cases (the therapy probably acts on more recent thrombi). Moreover, this therapy decreases the incidence of embolic events, although it increases the risk of hemorrages. Since the management of this preventive treatment in general medicine is very difficult, we hope that our health organization will establish Anticoagulation Clinics for the centralized management of this therapy. PMID- 9359052 TI - Afterload mismatch and relation between cardiac output and peripheral resistance. PMID- 9359054 TI - Leptospirosis in California sea lions (Zalophus californianus) stranded along the central California coast, 1981-1994. AB - Prevalence of leptospirosis was determined in California sea lions (Zalophus californianus) stranded live along the central California (USA) coast between January 1981 and December 1994. Clinical signs of renal disease were seen in 764 (33%) of 2338 animals examined; 545 (71%) of these 764 animals died, with similar gross lesions of nephritis. In silver impregnation stains of sections of formalin fixed kidney, numerous loosely coiled spiral organisms were observed. Leptospira pomona kenniwicki was cultured from four kidney samples in 1991. Epizootics of leptospirosis occurred in 1984, 1988, 1991, and 1994, and were more common in the autumn, typically affecting juvenile males. In 1991 and 1994, 47 animals sampled had antibody titers to L. pomona greater than 1:3200. In 1992, 20 animals sampled were seronegative, and in 1993 three of 20 animals sampled had low titers to L. pomona. PMID- 9359053 TI - Characterization of Borrelia spp. isolated from the tick, Ixodes tanuki and small rodents in Japan. AB - Spirochetes were isolated from the tick, Ixodes tanuki, as well as wood mice (Apodemus speciosus) and voles (Clethrionomys rufocanus and Eothenomys smithii), caught in Fukui, Tokushima, and Hokkaido, Japan, from 1991 to 1993. Spirochetes were characterized on the basis of protein profiles, reactivities with monoclonal antibodies (mAbs), Outer surface protein A gene (ospA) and Outer surface protein B gene (ospB) amplification analysis, rRNA gene and flagellin gene restriction fragment length polymorphism (RFLP) analysis, and DNA homology values. Protein profiles of all isolates were homologous and reacted with mAb to OspA, OspB, OspC, flagellin, and heat shock protein 60. The primer reactivity to ospA and ospB were different from those of Borrelia burgdorferi sensu stricto, B. afzelii, B. japonica, and B. garinii. Based on the DNA/DNA homology value and RFLP analysis of rRNA and flagellin gene, these Borrelia sp. isolates are a new group of B. burgdorferi sensu lato. The isolates from ticks and the host rodents were identical in these assays. We propose that these Borrelia sp. are adapted to I. tanuki and are maintained in these field rodents. PMID- 9359055 TI - The pathogenicity of Brucella suis biovar 4 for bison. AB - The pathogenicity of Brucella suis biovar 4 for bison (Bison bison) was evaluated by inoculation of 2.1 x 10(7) colony forming units (CFU) in 0.1 ml saline into the conjunctival sac of six pregnant cows. Six pregnant bison were inoculated with 1.27 x 10(7) CFU of Brucella abortus strain 2308 as a positive control. Bison were inoculated on 23 January 1992, and observed until calving or abortion after which they were euthanized, and necropsied. Bacteriological and histological examinations were conducted on lymph nodes, reproductive tract, mammary gland, and internal organs. Terminal serum samples from calves and cows were evaluated by card, rivanol precipitation, standard tube agglutination, cold complement fixation tube, indirect bison conjugated enzyme linked immunosorbent assay (ELISA), competitive ELISA, and particle-concentration fluorescence immunoassay. No clinical signs of brucellosis were seen in bison inoculated with B. suis biovar 4, and infection was found only in lymph nodes of two animals. There was no evidence of metastasis of this organism to the mammary gland or the reproductive tract. There were no detectable levels of antibodies to Brucella spp. in terminal blood samples taken from B. suis biovar 4-challenged bison. Brucella abortus was isolated from several tissues in all control bison. All B. abortus-challenged animals developed uterine infection and five developed mammary gland infection. Reproductive disease resulted in abortions in five B. abortus challenged bison and neonatal death in the remaining calf. Brucella suis biovar 4 does not appear to be pathogenic for bison. PMID- 9359056 TI - Susceptibility of Dall sheep (Ovis dalli dalli) to pneumonia caused by Pasteurella haemolytica. AB - We evaluated susceptibility of Dall sheep (Ovis dalli dalli) to bacterial pneumonia induced by two strains of Pasteurella haemolytica of domestic sheep origin by evaluating the sensitivity of blood neutrophils of eight Dall sheep to lysis by cytotoxins of P. haemolytica, and by intratracheal inoculation of three Dall sheep, two bighorn sheep (Ovis canadensis), and two domestic sheep with 3.7 x 10(6) or 2.5 x 10(7) colony forming units of P. haemolytica. Neutrophils from the Dall sheep were more sensitive to lysis by cytotoxins from supernatants of a P. haemolytica, biotype A, serotype 2 (A2), of domestic sheep origin, than were neutrophils from six bighorn sheep. This cytotoxic bacterium was the same isolate that was used for intratracheal inoculation of two Dall sheep and two domestic sheep. Inoculation of this cytotoxic P. haemolytica A2 resulted in fatal fibrinopurulent pleuropneumonia in the first Dall sheep within 24 hr of inoculation, and pneumonic lesions in the second Dall sheep before it was euthanized 52 hr after inoculation. This strain of P. haemolytica A2 did not cause respiratory disease when inoculated into two domestic sheep. A noncytotoxic strain of P. haemolytica; biotype T, serotype 3,4,10 of domestic sheep origin did not result in pneumonia in the third Dall sheep or two bighorn sheep. Prior to inoculation, P. haemolytica, biotype T isolates were obtained from all three Dall sheep, but none of these isolates was cytotoxic. At necropsy, cytotoxic P. haemolytica A2 was isolated from lungs and other tissues of the two pneumonic Dall sheep. Based on these results, we conclude that Dall sheep appear to be at least as sensitive as bighorn sheep to pneumonia caused by P. haemolytica A2 of domestic sheep origin. Because in vitro and in vivo results appear closely correlated in this and other studies, we believe with additional evaluation and standardization, neutrophil cytotoxicity tests may serve as a substitute for live animal challenges in future studies of pathogenic P. haemolytica in wild sheep. PMID- 9359058 TI - Mule deer (Odocoileus hemionus) and elk (Cervus elaphus) as experimental definitive hosts for Fascioloides magna. AB - In August 1992, six mule deer (Odocoileus hemionus hemionus) fawns and four elk (Cervus elaphus) calves (n = 2) or yearlings (n = 2) each were inoculated orally with 50, 250, or 2,000 metacercariae of the liver fluke Fascioloides magna to evaluate their potential to serve as definitive hosts. Animals were maintained for up to 403 days. Three mule deer each inoculated with 50 metacercariae survived the infection and shed eggs in feces; thus mule deer can function as definitive hosts for F. magna. The other three mule deer inoculated with 50 (n = 1) or 250 (n = 2) metacercariae died from fluke infection on days 91, 150, and 162 days postinoculation, respectively, and only immature F. magna were recovered. One elk calf inoculated with 2,000 metacercariae died from fluke infection 44 days after inoculation. The remaining three elk, each inoculated with 250 metacercariae, survived infection, and two of the three shed eggs in feces. The third elk contained only one immature F. magna at necropsy. The prepatent period in mule deer and elk was approximately 6 to 7 months. PMID- 9359057 TI - Experimental contact of bighorn sheep (Ovis canadensis) with horses and cattle, and comparison of neutrophil sensitivity to Pasteurella haemolytica cytotoxins. AB - Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P. haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophil death in vitro) to bighorn sheep neutrophils, and four of 10 P. haemolytica biotype A isolates were cytotoxic to neutrophils of cattle; P. haemolytica biotype T (= Pasteurella trehelosi) isolates were noncytotoxic to neutrophils of bighorn sheep and cattle. When six bighorn sheep were pastured with three horses, only P. haemolytica biotype T isolates were recovered from the bighorn sheep throughout the study; Pasteurella spp. organisms were not isolated from the three horses. At initiation of a study where five bighorn sheep were pastured with three cattle, P. haemolytica biotype A, serotype 1, 2 was isolated from all three cattle, and only P. haemolytica biotype T isolates were recovered from the bighorn sheep. One bighorn sheep died in each of the horse and cattle copasturing experiments. Pasteurella haemolytica was not isolated from the bighorn sheep which died in the horse copasturing experiment. A noncytotoxic P. haemolytica biotype A, serotype 2 was isolated at necropsy from the bighorn which died in the cattle contact experiment. Based on these experiments, we believe bighorn sheep and horse association would not be detrimental to bighorns due to P. haemolytica induced pneumonia. PMID- 9359059 TI - Parasitism of Gobius bucchichii Steindachner, 1870 (Teleostei, Gobiidae) in protected and unprotected marine environments. AB - We collected 396 Gobius bucchichii, Steindachner, 1870 (Teleostei, Gobiidae) in and around the marine reserve of Cerbere-Banyuls, in the southeast of France, between March and July 1994. Five species of adult parasites were found: one acanthocephalan, Acanthocephaloides propinquus Dujardin, 1845 (Acanthocephala, Arhythmacanthidae); one nematode, Cucullanus sp. (Nematoda, Cucullanidae); and three species of digenetic trematodes, Helicometra sp. (Digenea, Opecoelidae), Derogenes sp. (Digenea, Hemiuridae) and Deretrema scorpaenicola Bartoli, 1990 (Digenea, Zoogonidae). Fishes collected in a protected area were on average, larger, older, had a higher percentage of regenerated scales, and harbored more parasites. PMID- 9359060 TI - Meningeal worm in experimentally-infected bighorn and domestic sheep. AB - In the first (July 1989) of two experiments, each of three bighorn sheep (Ovis canadensis) and three domestic sheep, respectively, was exposed to 25, 150, or 300 infective third-stage larvae (L3) of the meningeal worm, Parelaphostrongylus tenuis. Two bighorn sheep had temporary mild paresis and lumbar weakness; one developed paralysis and died suddenly 32 days after exposure. Adult P. tenuis were found deep within the brain and spinal cord of the one latter sheep. A generalized inflammatory response, characterized by subdural lymphoid aggregations adjacent to spinal nerve roots, was seen in the spinal cord of most domestic and bighorn sheep. In the second experiment (September 1990), each of six domestic sheep lambs and five white-tailed deer (Odocoileus virginianus) fawns was exposed to a single dose of 15 to 125 L3 of meningeal worm. Clinical signs were seen in only one sheep; it was dull and depressed. No worms were found in this sheep. One dead adult meningeal worm was found on the brain of another sheep. First-stage larvae and adult meningeal worms were found in all deer. PMID- 9359061 TI - Spontaneous cryptosporidiosis in captive white-tailed deer (Odocoileus virginianus). AB - In August 1994, cryptosporidiosis was diagnosed in a diarrheic fawn from a captive white-tailed deer (Odocoileus virginianus) herd maintained for research purposes at The University of Georgia's Warnell School of Forest Resources in Athens, Georgia (USA). From June through August 1995, 11 captive female white tailed deer were housed in individual barn stalls where they gave birth to 18 fawns. Feces collected at 2 or 3 day intervals from the 18 neonatal fawns for at least 21 days and from 11 adult females once from 1 to 30 days before fawns were born and on three to 12 occasions after their birth were examined for oocysts of Cryptosporidium spp. Feces from all animals appeared normal throughout the period of examination. Oocysts morphologically indistinguishable from those of Cryptosporidium parvum were detected intermittently in the feces of one adult female from 1 to 25 days after parturition and in the feces of her fawn from 11 to 22 days of age. Oocysts also were detected intermittently in feces from twin fawns from 9 to 20 days of age, but not from their mother. Oocysts from deer were infectious for neonatal mice as determined histologically, and for calves as determined by clinical signs and excretion of oocysts. PMID- 9359062 TI - Detection of agglutinating antibodies to Toxoplasma gondii in sera from free ranging eastern barred bandicoots (Perameles gunnii). AB - Sera from 150 eastern barred bandicoots (Perameles gunnii) were collected from two study sites in southern Tasmania between 1992 and 1995. Samples were tested for antibodies to the protozoan parasite, Toxoplasma gondii, using formalin treated tachyzoites as the antigen in direct (DAT) and modified agglutination tests (MAT). Cut-off titers were set based on confirmed cases of toxoplasmosis in this species. A total of 133 animals (89%) were classified as negative, seven (4.6%) had suspicious reactions, and 10 (6.7%) were diagnosed as positive. Five of the 10 positive animals were not retrapped after initial seroconversion; another three animals recorded high MAT titers on two consecutive bleedings, three months apart. Of the remaining two sero-positive bandicoots, one was found dead in a trap and generalized toxoplasmosis was diagnosed at necropsy, while the other animal had central nervous system disabilities consistent with toxoplasmosis but was accidently released and never recaptured. Based on these findings we propose that eastern barred bandicoots are likely to be highly susceptible to primary T. gondii infection. PMID- 9359063 TI - Monitoring of Culicoides spp. at a site enzootic for hemorrhagic disease in white tailed deer in Georgia, USA. AB - Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) were monitored at a Georgia (USA) site where epizootic hemorrhagic disease (EHD) and bluetongue (BT) viruses are enzootic among white-tailed deer (Odocoileus virginianus). Collections were made using a captive white-tailed deer and light traps from June 1993 through November 1994. We collected 210,482 females from the captive deer during morning and evening periods. Predominant species were C. lahillei (73%), C. stellifer (16%), C. biguttatus (6%), C. niger (3%), C. spinosus (2%), and C. paraensis (0.2%). Other species were C. venustus, C. obsoletus/sanguisuga, C. haematopotus, C. guttipennis, and C. arboricola, which together represented < 0.1% of the specimens collected. No C. variipennis, a known vector of EHD and BT viruses, were collected from the deer. An estimated 953,299 females were collected in 695 light-trap nights. The most common species in light-trap collections were C. spinosus (45%), C. biguttatus (27%) and C. stellifer (24%). Culicoides variipennis was rare in the light-trap samples, representing < 0.01% of the total collections. There was serological evidence from hunter-killed deer that local deer were infected with EHD and BT viruses during the study, particularly during 1994. A primary suspect vector was C. lahillei, which attacked the bait deer in large numbers during the summer and early fall of both 1993 and 1994. Based on their seasonality, relative abundance, and host-seeking activity, C. stellifer and C. spinosus also were considered as possible vectors. However, virus isolation attempts on 113,716 Culicoides, including 62,530 C. lahillei and 32,769 C. stellifer, were negative. PMID- 9359064 TI - Weights, hematology, and serum chemistry of seven species of free-ranging tropical pelagic seabirds. AB - I established reference values for weight, hematology, and serum chemistry for seven species of free-ranging Hawaiian tropical pelagic seabirds comprising three orders (Procellariiformes, Pelecaniformes, Charadriiformes) and six families (Procellariidae, Phaethontidae, Diomedeidae, Sulidae, Fregatidae, and Laridae). Species examined included 84 Hawaiian darkrumped petrels (Pterodoma phaeopygia), 90 wedge-tailed shearwaters (Puffinus pacificus), 151 Laysan albatrosses (Diomedea immutabilis), 69 red-footed boobies (Sula sula), 154 red-tailed tropicbirds (Phaeton rubricauda), 90 great frigatebirds (Fregata minor), and 72 sooty terns (Sterna fuscata). Hematocrit, total plasma solids, total and differential white cell counts, serum glucose, calcium, phosphorus, uric acid, total protein, albumin, globulin, aspartate aminotransferase and creatinine phosphokinase were analyzed. Among and within species, hematology and chemistry values varied with age, sex, season, and island of collection. Despite this variation, order-wide trends were observed. PMID- 9359065 TI - Quadricuspid aortic valve and single coronary artery in a greater white-toothed shrew, Crocidura russula. AB - An adult greater white-toothed shrew (Crocidura russula) had both a quadricuspid aortic valve and a single coronary artery arising from the aorta. The shrew was caught on 10 May 1994 in the environs of Malaga, southern Spain. Both congenital anomalies may be potential causes of cardiac dysfunction, but apparently produced no significant cardiac complication in the shrew. This is the first report of a quadricuspid aortic valve in a wild-living mammal. PMID- 9359066 TI - Cryptorchidism and delayed testicular descent in Florida black bears. AB - Retained testes were found in 11 (16%) of 71 black bears (Ursus americanus) examined over a 3-year period in Florida (USA). Four of the 11 bears were older than one year and weighed more than 32 kg; therefore, they were considered to be cryptorchid. The remaining seven bears may have had delayed testicular descent due to their apparent normal immature development. This is the first known published report of the prevalence of cryptorchidism and apparently normal delayed testicular descent in a black bear population. PMID- 9359067 TI - Hemorrhagic gastritis in free-living rodents in Idaho. AB - Between February 1992 and March 1994, four species of rodent from the Snake River Birds of Prey Area near Boise, Idaho (USA) were necropsied. Hemorrhagic gastritis was observed in 16 of 131 Townsend's ground squirrels (Spermophilus townsendii), one of 11 Ord's kangaroo rats (Dipodomys ordii) and the one Great Basin pocket mouse (Perognathus parvus) evaluated. No lesions were observed in 14 white-footed deer mice (Peromyscus maniculatus). Tissue from one Townsend's ground squirrel was negative for Helicobacter sp.-like bacteria. PMID- 9359068 TI - Lymphosarcoma in a raccoon (Procyon lotor). AB - A case of lymphosarcoma in a captive adult female raccoon (Procyon lotor) from northeastern Pennsylvania (USA) was observed in 1991. Prior to its death the raccoon had lost weight. At necropsy the carcass was in poor body condition and had pale mucous membranes. The thoracic and abdominal lymph nodes were enlarged, soft, and pale tan. Microscopically, there was effacement of normal lymph node architecture by sheets of mononuclear cells. These were well-differentiated small lymphocytes with distinct cell borders. Nuclei of these cells were darkly stained and mitotic figures were frequently seen. Similar but lesser numbers of neoplastic cells were seen in the parenchyma of liver, spleen, and the pancreas. Since the neoplasm involved several organs, we propose that the condition was of multicentric origin. Gross lesions, histopathologic findings and the organs involved differed from a previously described case of lymphosarcoma in a raccoon. PMID- 9359069 TI - Polycystic kidney disease in a raccoon (Procyon lotor). AB - During March 1988, a case of bilateral polycystic kidney disease (PKD) occurred in an aged raccoon (Procyon lotor) at a zoo in Wilmington, Delaware (USA). Prior to its death, the raccoon had no clinical signs. On necropsy there was bilateral enlargement of kidneys which, on cut sections, had many variable sized fluid filled cystic cavities. Endometrial hyperplasia with presence of multiple variable-sized cysts were also seen in the uterus of this raccoon. Microscopical examinations were characteristic of an end-stage renal failure due to PKD. Neither PKD nor cystic endometerial hyperplasia appears to have been previously described in this species. PMID- 9359070 TI - Evaluation for malignant hyperthermia susceptibility in black-tailed deer. AB - To investigate a possible link of malignant hyperthermia to capture myopathy, between June 1990 and July 1993 we anesthetized four black-tailed deer (Odocoileus hemionus columbianus) and challenged them with halothane and succinylcholine. Halothane had no significant effect on oxygen consumption. Succinylcholine significantly (P < 0.05) increased cardiac output (mean +/- SD), from 2.94 +/- 1.05 l/min to 5.26 +/- 1.79 l/min, and oxygen consumption, from 5.5 +/- 2.1 ml/kg/min to 10.1 +/- 2.9 ml/kg/min. Muscle biopsy specimens tested for malignant hyperthermia susceptibility responded normally to halothane and caffeine. We conclude that these deer did not experience malignant hyperthermia; suggesting no link to capture myopathy. PMID- 9359071 TI - Respiratory and pharyngo-esophageal iridovirus infection in a gopher tortoise (Gopherus polyphemus). AB - A free-living adult male gopher tortoise (Gopherus polyphemus) was found on Sanibel Island, Florida (USA), on 18 February 1992 with signs of upper respiratory disease. On necropsy after euthanasia on 27 February 1992, severe, extensive necrotizing ulcerative tracheitis, multifocal necrotizing pneumonia, and multifocal necrotizing ulcerative pharyngitis and esophagitis were observed. Large ovoid to round intracytoplasmic basophilic inclusions, which appeared to displace the nucleus to the cell periphery, occurred within degenerate and necrotic epithelial cells of the above tissues. On transmission electron microscopy of formalin-fixed trachea and lung, intracytoplasmic viral particles were observed within necrotic cells in the tracheal lumen and epithelial cells of the lung. Most infected cells also had a roughly spherical granular cytoplasmic inclusion that contained clusters of viral particles. Viral particles had an electron dense spherical to icosahedral core surrounded by a less electron dense icosahedral capsid. Mature extracellular virions were surrounded by an envelope and were 150 to 220 nm in diameter. Virions and cytoplasmic inclusions were morphologically similar to those of the Family Iridoviridae. PMID- 9359072 TI - Brief characterization of muskrat (Ondatra zibethicus) immunoglobulin G (IgG) separated from serum on protein A. AB - Muskrat (Ondatra zibethicus) immunoglobulin fraction was separated from whole serum by Protein A Sepharose chromatography. In serum electrophoresis, this fraction had a gamma motility; when electrophoresed on a polyacrylamide gel with sodium dodecyl sulfate it resolved into two protein bands of approximately 52 and 25 kilodaltons, respectively. These bands were consistent with molecular weights of known heavy and light chains of immunoglobulin G (IgG) in other closely related species. Furthermore, the putative muskrat immunoglobulins had a strong cross-reactivity with mouse IgG1, IgG3, and kappa chain in an enzyme-linked immunosorbent assay. We propose, that the proteins bound to the Protein A Sepharose represent muskrat immunoglobulins of the IgG class. PMID- 9359074 TI - Hemorrhagic disease in white-tailed deer in Texas: a case for enzootic stability. AB - Although antibodies to viruses in both the epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) sero-groups have been reported from white tailed deer (Odocoileus virginianus) in Texas (USA), there are few reports of hemorrhagic disease (HD) in these populations. To understand the extent and diversity of exposure to the North American EHDV and BTV serotypes in these deer populations, we serologically tested 685 white-tailed deer collected from November 1991 through March 1992 throughout their range in Texas. Overall, 574 (84%) of deer had antibodies to EHDV or BTV. Prevalence estimates varied according to ecological region, from 57% in the Gulf Prairies to 100% in the northwest Edwards Plateau. Based on serum neutralization tests, the deer had evidence of previous exposures to multiple EHDV and BTV serotypes, with evidence of exposure to two to five serotypes detected in each ecological region. The apparent lack of HD in relation to this high antibody prevalence cannot be explained, but may be related to enzootic stability in which a near perfect host virus relationship exists. PMID- 9359073 TI - Isolation of Mycoplasma felis from a serval (Felis serval) with severe respiratory disease. AB - We report cytologic observations and isolation of Mycoplasma felis in September 1992 from the lower respiratory tract of a 3-week-old captive serval (Felis serval) cub with pneumonia, in Florida (USA). Septic, neutrophilic inflammation with a large, monomorphic population of unique, pleomorphic, intracellular and extracellular rods was diagnosed from a transtracheal wash. Mycoplasma felis was the only bacterium isolated in significant numbers from the transtracheal wash. PMID- 9359075 TI - Parelaphostrongylus tenuis on Wassaw Island, Georgia: a result of translocating white-tailed deer. AB - Meningeal worms (Parelaphostrongylus tenuis) were found in each of five white tailed deer (Odocoileus virginianus) examined from Wassaw Island, Chatham County, Georgia, in September 1993. This represents the first reported occurrence of the parasite on a southeastern barrier island and extends its geographic distribution approximately 140 km beyond the nearest known infected mainland deer population. According to an anecdotal account, six white-tailed deer were imported from Pennsylvania and released on Wassaw Island in 1905 or shortly thereafter. Based on its absence elsewhere along the southeastern coast from North Carolina to Louisiana and its high prevalence in Pennsylvania, the enzootic focus of P. tenuis on Wassaw Island was attributed to translocation of infected deer. PMID- 9359076 TI - Porrorchis hylae (Johnston, 1914) (Acanthocephala: Plagiorhynchidae: Porrorchinae) in Lialis burtonis, Gray 1835 (Sauria: Pygopodidae); a new paratenic host record. AB - During necropsy of a Burton's snake lizard (Lialis burtonis Gray 1835) that was imported from Australia in January 1994, juvenile acanthocephalans were recovered from the mesentery and small bowel serosa which were identified as Porrorchis hylae (Johnston, 1914). This find represents a new paratenic host record for the species, and is recorded in a reptile native to Australia for the first time. PMID- 9359078 TI - Antibody responses to Psoroptes sp. mites in Dall sheep (Ovis dalli). AB - We determined that antibody responses to Psoroptes sp. mites were not present in 403 of 407 sera samples collected opportunistically from 1979 through 1991 from Dall sheep (Ovis dalli) from five locations in Alaska, USA (Eastern Arctic, n = 61; Central Arctic, n = 15; Western Interior, n = 122; Central Interior, n = 63; Eastern Interior, n = 146). Test values for four samples exceeded the positive cutoff value for the immunoassay, but exposure to mites could not be confirmed since the 95% confidence interval for true prevalence ranged from 0 to 2.3%. Therefore, we concluded that these were probably false positive results. Our analysis, coupled with the lack of previous reports of mites or lesions in Dall sheep or other Alaskan ungulates, provided indirect evidence that Psoroptes sp. are not enzootic in Dall sheep in Alaska. In contrast, Psoroptes sp. have been reported in bighorn sheep (Ovis canadensis) and other wild ungulate populations from southern Canada to Mexico. These findings are compatible with the hypothesis that Psoroptes sp. were introduced into North America with imported domestic sheep and were not introduced by ancestral wild sheep. PMID- 9359077 TI - Coccidioidomycosis in free-living California sea lions (Zalophus californianus) in central California. AB - Coccidioidomycosis is described in seven California sea lions (Zalophus californianus) admitted to The Marine Mammal Center, Sausalito, California (USA), between January 1986 and December 1994. Diagnoses were confirmed by histology in all seven cases, culture in three cases, and serology in one case. These are believed to be the first published cases of coccidioidomycosis in free-ranging California sea lions. PMID- 9359079 TI - Effect of ectoparasite removal procedures on recapture of Microtus californicus. AB - We tested the null hypothesis that anesthetizing meadow voles (Microtus californicus) and brushing them vigorously for ectoparasites would have no effect on the later recapture of these voles. Voles were trapped from 6 December 1993 to 10 January 1994 at Faye Slough Wildlife Area near Eureka, Humboldt County, California (USA). Alternate trapped voles were anesthetized with ethyl ether and brushed vigorously for ectoparasites. There was no significant difference in the frequency of recapture nor in the time to first recapture between those voles anesthetized and brushed, compared to control animals. PMID- 9359080 TI - The role of diagnostic statistics in medicinal chemistry. PMID- 9359081 TI - Alpha-adrenergic approach in the medical management of benign prostatic hyperplasia. PMID- 9359082 TI - Molecular targets for the rational design of antiepileptic drugs and related neuroprotective agents. PMID- 9359083 TI - Use of an open stereotactic ring for neurosurgical procedures. AB - We describe an open ring as a new design to the Zamorano-Dujovny (Z-D) stereotactic unit. The titanium base ring has an opening of 45 degrees that can be located in any chosen position. Imaging studies such as computed tomography, X ray, positron emission tomography, digital angiography, and digital substraction angiography can be performed with the open stereotactic ring for multimodality image localization. Preoperatively and intraoperatively, this open design provides the anesthesiologist with an unobstructed pathway for airway management. During the surgical procedure, it facilitates approach to any intracranial lesion, including orbitozygomatic, combined supra-infratentorial, and others. During awake craniotomies it not only allows for easy airway management, but also provides good access to the patient's face for intraoperative evaluation of speech and visual functions. Accuracy and reliability of this unit were similar to results obtained with the original circular ring. This system can be used in conventional stereotaxis with the Z-D arc, as well as a reference for intraoperative registration with any digitizer system. The open stereotactic unit is a relatively inexpensive, reliable, and easy-to-use solution for resections using conventional stereotaxis or interactive image guidance in any intracranial site. PMID- 9359085 TI - Stereotactic radiosurgery of skull base meningiomas. AB - Between April 1992 and February 1996, 97 patients with skull base meningiomas were treated at our department. The age of these patients ranged from 10 to 80 years. The male/female ratio was 1/2. Fifty-three of these patients had primary open surgery for partial removal or recurrent growth and subsequent radiosurgical treatment. Radiosurgery was performed as a primary treatment in 44 patients. The mean tumor volume was 13.7 cm3 (range: 0.8-82 cm3). These tumor volumes could be covered by mean isodose volumes of 45% (range: 20-70%) and were treated by a mean dose of 13.8 Gy (range: 7-25 Gy) at the tumor border. Six patients underwent radiosurgery with a staged treatment protocol with 4.6-6 months interval. In 78 patients, a total of 102 follow-up scans were available. The remaining 19 patients have not been included in the post-radiosurgical evaluation since the observation time was either too short or the patients were lost for follow-up. The mean interval between gamma knife treatment and last follow-up scan was 18.5 months, with a range from 6 to 46 months. Follow-up imaging (CT, MRI or both) revealed a decreased volume of the tumor in 31 cases (40%). In 44 cases (56%), tumor progression was stopped, and in 3 cases (4%) increased tumor volumes could be observed. In 8 cases marked central tumor necrosis was seen. Neurological follow-up examinations in 76 patients showed a stable neurological status in 71%, ameliorated status in 24% and worsening in 5% of the patients. PMID- 9359084 TI - Stereotactic neurosurgery planning with 3-D spiral CT-angiography. AB - OBJECTIVE: To assess the feasibility and value of spiral CT angiography of the brain vessels for the planning of neurosurgical stereotactic interventions. MATERIAL AND METHODS: Fourty-two patients harboring cerebral lesions underwent spiral CT angiography prior to stereotactic biopsy. Thin spiral CT slices with a collimator slice thickness of 1 mm and a pitch of 1 were used. Multiplanar reconstructions and maximum intensity projections (MIP) were obtained as well as 3-D tissue definition. RESULTS: There was a sufficient visualization of vessels and of their relationship to the lesion. Tumor neovascularization was clearly demonstrated. Arteries could be shown separately. Stereotactic coordinates of targets were chosen at a safe distance from the vessels and the simulation of tarjectories using the cine loop was made possible. In three cases the presence of a pathological vascularization warned against a stereotactic biopsy. CONCLUSION: Spiral CT angiography seems to yield enough topographical information for the accurate planning of stereotactic surgery for brain lesions. CT angiography with the helical technique is rapid and less invasive than digital subtraction angiography. PMID- 9359086 TI - Orbital roof craniotomy via an eyebrow incision: a simplified anterior skull base approach. AB - Utilizing the conceptual combination of brain protective skull base surgery and minimalism, a conventional frontal craniotomy for tumors in the subfrontal and parasellar regions is modified to an orbital roof craniotomy. Through a 4 to 5 centimeter (cm) long eyebrow incision an orbital roof craniotomy (measuring 2 cm by 3 cm), including the supraorbital arch, is made as a single piece bone flap. The orbital roof is opened up to the supraorbital fissure and to the optic canal by additional removal of the bone in the orbital roof. This will expose the globe and the orbitofrontal dura mater. When the dural incision is made at the orbital portion of the dura mater, the orbital contents are retracted by tack-up sutures. The tumor is removed utilizing the orbital space rather than the intracranial space. Brain retractors are not necessary and are not used to execute the tumor resection. This technique has been used in three patients with craniopharyngiomas, seven patients with meningiomas, and one patient with a subfrontal teratoma. Gross total resection was achieved in three patients with craniopharyngiomas and in five patients with subfrontal or parasellar meningiomas. Subtotal resection of the tumor was achieved in two patients with recurrent meningiomas and in the patient with a subfrontal teratoma. The surgeon's operating space through this exposure was sufficiently ample to achieve the goals of the operation. The direct eyebrow incision provides an additional vital working space with a width of more than 1 cm at the skull base by eliminating the scalp flap which a coronal incision employs. The surgical technique is described with a report of 11 cases. PMID- 9359087 TI - Experimental use of semiconductor diode laser for neuroendoscopic surgery. AB - The authors demonstrate an experimental use of the newly developed high power aluminium-gallium-arsenide (AlGaAr) diode laser (DIOMED 25, Olympus Optical Company, Tokyo, Japan). The unit consists of a compact body and fiberoptic probes with small accessories. There are two types (contact and non-contact) of probes. Tissue effects on rat liver, femoral artery, and brain tissue were examined. Adding that, we measured the thermal changes on the liver surface produced by the laser beam with a thermography system. For coagulation with the contact probe, 5 or 7 W was adequate but 10 W was too excess because of tissue adhesion. For cutting, low absorption of the laser in less vascularized tissue like brain white matter provided a deeper tissue damage compared with more vascularized tissue. The temperature at the center reached over 100 degrees C during 10 seconds after laser treatment with the cutting probe. These findings suggest that this system proved to be a good candidate for endoscopic hemostasis and cutting with meticulous maneuver. PMID- 9359088 TI - Asymmetric hydrocephalus: safe endoscopic perforation of septum pellucidum: technical note. AB - Asymmetric and/or loculated hydrocephalus can be treated with endoscopic septum fenestration to avoid or to simplify a shunt (1,2). In asymmetric lateral ventricles the septum pellucidum is dislocated to the opposite side and may even be in contact with the lateral wall of the contralateral ventricle (i.e., the thalamus). Perforating the septum with a catheter or a laser beam may damage the underlying tissue. The authors show a safe perforation technique: the septum is pulled towards the tip of the endoscope to enlarge the underlying space. Now the catheter can perforate the septum without the risk of damage to the underlying tissue. PMID- 9359089 TI - Endoscopic treatment of brain abscess in children. AB - Three children with intracerebral abscesses were treated endoscopically. Two of the treated abscesses were located in the left temporal lobe and one in the right parietal lobe. The presenting symptoms included headaches, seizures, hemiparesis and signs of infection. Burr hole craniotomy, insertion of a peelaway sheath, obtaining of a specimen, introduction of endoscope, and complete irrigation under view was performed. After this a draining catheter was positioned in the abscess. All three abscesses grew multiple organisms. The patients received longstanding intravenous treatment with antibiotics. The follow-up period in this group ranges between 5 and 32 months. The initial neurological deficits were relieved in all three patients. The follow-up MRI studies revealed minor residual changes without evidence of significant sequelae. Neuroendoscopic treatment of brain abscesses has additional advantages compared to stereotactic aspiration or more complete drainage and lavage. PMID- 9359090 TI - Intrasellar gangliocytoma resembling pituitary adenoma. AB - Cushing's disease resulting from intrasellar gangliocytomas is very rare and only three cases have been reported to date. All of the cases were female. We present a fourth case of Cushing's disease resulting from intrasellar gangliocytoma. Computed tomography and magnetic resonance imaging scans showed a pituitary macrotumor with suprasellar extension. A greenish-gray colored homogenous tumor was subtotally removed by a transcranial approach. Histological diagnosis was gangliocytoma. To the best of our knowledge, this is the first reported instance of a tumor causing Cushing's disease in a man in the absence of a pituitary adenoma component. PMID- 9359092 TI - Drug discovery in the postgenomic era. PMID- 9359091 TI - Computer navigational microscope for minimally invasive neurosurgery. AB - Frameless navigational devices have recently undergone an evoluation to the point where they are being used more efficiently in the clinical setting. Most of the present-day systems utilize some form of mechanical arm to correlate registered points in space to the computer terminal. However, the articulated arm can be cumbersome and is an additional obstacle in the surgical field. To avoid this problem some groups have elected to transmit registered points and probe position using ultrasound or electromagnetic fields. However, in using magnetic fields, ferromagnetic metals can interfere with accuracy. On the other hand with ultrasonic digitizers accuracy is dependent on variable factors such as humidity, local temperature, and positioning of the emitters. In response to some of these difficulties inherent with frameless stereotaxy, the MKM system was developed. Essentially it is an optically based system in which the CT and/or MRI data are superimposed three-dimensionally onto the surgical field as seen through the microscope using head-up display technology. It requires no opto-kinetic link such as a headframe, mechanical arm, ultrasonic, and magnetic field. We have used the MKM microscope system to guide 18 neurosurgical procedures. This report illustrates the advantages of this system when combined with keyhole surgery to provide accurate excision of brain lesions while preserving normal tissue. PMID- 9359094 TI - Profit margins and epistemology. PMID- 9359093 TI - The challenges of genome sequence annotation or "the devil is in the details". PMID- 9359095 TI - General electric goes to the mat in the PCB wars. PMID- 9359096 TI - From syntax to semantics. PMID- 9359097 TI - Debunking hCG. PMID- 9359098 TI - Carbohydrates down, but not out. PMID- 9359099 TI - Thalidomide checks might have saved Redux. PMID- 9359100 TI - Enbrel's phase III reinforces prospects in RA. PMID- 9359101 TI - On target with tumor blood vessel markers. PMID- 9359102 TI - Hormone peptidomimetics: seeing double. PMID- 9359103 TI - Repelling plant pathogens with ribonuclease. PMID- 9359104 TI - Growth hormone technology develops new twist. PMID- 9359106 TI - Finding a new language for bioinformatics. PMID- 9359105 TI - Getting the right drug into the right patient. PMID- 9359107 TI - The simplicity of complex MACs. AB - The development of mammalian artificial chromosomes (MACs) would be useful for biotechnology and biomedicine, including their use in functional genomics, transgenic animals and gene therapy. By analogy to large cloning systems in microorganisms, MACs may be engineered using endogenous chromosomal elements such as the yeast-based artificial chromosomes (YACs), or exogenous extra-chromosomal components derived from viruses and other cellular parasites such as the bacterial-based artificial chromosomes (BACs) and p1 artificial chromosomes (PACs). PMID- 9359108 TI - Increased potency of an erythropoietin peptide mimetic through covalent dimerization. AB - We have synthesized a chemically defined, dimeric form of an erythropoietin mimetic peptide (EMP) that displays 100-fold increased affinity for the erythropoietin receptor (EPOR) and correspondingly elevated potency in cell-based assays and in mice. The dimeric EMP1 was synthesized using a C-terminal lysine residue as a branch point. A beta-alanine residue was coupled to the main-chain (alpha) amino group of the lysine residue in order to provide a pseudosymmetrical scaffold where both the side-chain and main-chain were of approximately equal length. Using an orthogonal protection system, independently disulphide-cylized EMP1 moieties were synthesized upon this scaffold. The proposed mechanism of increased potency of the dimer over the parental compound EMP1 is consistent with the structure of a cocrystal of EMP1 and the extracellular domain of the EPOR in which a noncovalent peptide dimer is seen spanning the cleft between two molecules of the EPOR extracellular domain. PMID- 9359109 TI - Structure-based design and characterization of exocyclic peptidomimetics that inhibit TNF alpha binding to its receptor. AB - Exocyclic small peptidomimetics corresponding to three critical binding sites of tumor necrosis factor (TNF)-receptor(I) have been designed based on atomic features deduced from the crystal structures of TNF alpha and the TNF beta/TNF receptor(I) complex and a model of an anti-TNF alpha monoclonal antibody. TNF alpha antagonistic activities were evaluated by binding assays using soluble receptor or intact receptor on cells as well as an apoptosis/cytotoxicity assay. The most critical interaction site for rational design of peptidomimetics was localized to the loop1/domain3 of the TNF-receptor. The best antagonist showed 5 microM inhibition in the binding assay. Biologically, the mimetics inhibited TNF alpha-mediated apoptosis. PMID- 9359110 TI - Targeting by affinity-matured recombinant antibody fragments of an angiogenesis associated fibronectin isoform. AB - The oncofetal fibronectin (B-FN) isoform is present in vessels of neoplastic tissues during angiogenesis but not in mature vessels. B-FN could therefore provide a target for diagnostic imaging and therapy of cancer. Phage display libraries have been used to isolate human antibody fragments with pan-species recognition of this isoform. We describe the use of these fragments in nude mice to target an aggressive tumor (grafted F9 murine teratocarcinoma). Imaging in real time was done by infrared photodetection of a chemically coupled fluorophore. The targeting was improved by use of affinity-matured fragments with low kinetic dissociation rates (koff = 1.5 x 10(-4) s-1) and also by engineering dimeric fragments via a C-terminal amphipathic helix. PMID- 9359111 TI - Creating a bifunctional protein by insertion of beta-lactamase into the maltodextrin-binding protein. AB - Hybrid proteins were generated by inserting the penicillin-hydrolyzing enzyme, TEM beta-lactamase (Bla), into the maltodextrin-binding protein (MalE). The inserted Bla was functionally accommodated by MalE when it was placed within permissive sites. The maltose binding and penicillinase activities of purified hybrids were indistinguishable from those of the wild-type MalE and Bla proteins. Moreover, these hybrids displayed an additional unexpected property: maltose stabilized the active site of inserted Bla. PMID- 9359112 TI - A protein particle vaccine containing multiple malaria epitopes. AB - Ty virus-like particles consist of a single protein species that can be produced in yeast. Recombinant Ty-VLPs carrying a string of up to 15 defined cytotoxic T lymphocyte (CTL) epitopes from Plasmodium species prime protective CTL responses in mice following a single administration without adjuvant. Effective processing of epitopes from the string was demonstrated in vitro and in vivo and was not affected by flanking sequences. PMID- 9359113 TI - Enhanced growth by ectopic expression of growth hormone releasing hormone using an injectable myogenic vector. AB - Ectopic expression of a truncated growth hormone-releasing hormone (GHRH) from muscle tissues by a myogenic plasmid DNA vector directed by the skeletal alpha actin promoter, pSK-GHRH, results in growth hormone (GH) secretion. Skeletal muscle-secreted GHRH is biologically active. The application of conditioned media harvested from pSK-GHRH transfected muscle cells to cultured pig primary anterior pituitary cells elicits GH release. A single intramuscular injection of 100 micrograms pSK-GHRH DNA elevates serum GH levels threefold to fourfold for up to 2 weeks, enhancing liver IGF-1 gene expression and increasing body weight approximately 10%. PMID- 9359114 TI - Transgenic potato expressing a double-stranded RNA-specific ribonuclease is resistant to potato spindle tuber viroid. AB - We have produced transgenic potato lines expressing the yeast-derived double stranded RNA-specific ribonuclease pac1. Five lines of pac1 potato (Solanum tuberosum L., cultivar Russet Burbank) challenged with potato spindle tuber viroid (PSTVd) suppressed PSTVd infection and accumulation. All of the progeny potato tubers produced by resistant plants were also free of PSTVd. Because the pac1 gene product digested PSTVd in vitro, double-stranded regions in PSTVd molecule and/or replicative intermediates may be targeted by pac1 gene product in the transgenic potato plant. PMID- 9359115 TI - Transformation of wheat with high molecular weight subunit genes results in improved functional properties. AB - The high molecular weight (HMW) subunits of wheat glutenin are major determinants of the elastic properties of gluten that allow the use of wheat doughs to make bread, pasta, and a range of other foods. There are both quantitative and qualitative effects of HMW subunits on the quality of the grain, the former being related to differences in the number of expressed HMW subunit genes. We have transformed bread wheat in order to increase the proportions of the HMW subunits and improve the functional properties of the flour. A range of transgene expression levels was obtained with some of the novel subunits present at considerably higher levels than the endogenous subunits. Analysis of T2 seeds expressing transgenes for one or two additional HMW subunits showed stepwise increases in dough elasticity, demonstrating the improvement of the functional properties of wheat by genetic engineering. PMID- 9359116 TI - Recovery of homogeneous and functional beta 2-adrenergic receptors from extracellular baculovirus particles. AB - Expression in baculovirus-infected insect cells allows sufficient production of G protein coupled receptor for structural studies. An important drawback of this expression system comes from the presence of unprocessed and biologically inactive receptors that have to be eliminated during receptor purification steps. We show that viral particles released from Sf9 cells infected with a recombinant baculovirus coding for the human beta 2-adrenergic receptor (beta 2AR) cDNA contain glycosylated and biologically active beta 2AR. In addition, post translational modifications known to modulate receptor activity were found to occur in these particles. PMID- 9359117 TI - Catalytic antibodies. Some companies are taking an active interest in this promising technology. PMID- 9359118 TI - Psychosensory symptoms in bipolar disorder. AB - This study investigated psychosensory symptoms and their relationship to retrospective and prospective courses of illness, as well as therapeutic outcomes, in patients with bipolar disorder. Using the Silberman-Post Psychosensory Rating Scale (SP-PSRS), psychosensory symptoms were assessed in 51 patients who met Diagnostic and Statistical Manual, 3rd Edition-Revised (DSM-III R) criteria for bipolar disorder and in 39 healthy, normal controls. Patients with bipolar disorder were enrolled in a 3-year, double-blind, randomized study comparing the prophylactic efficacy of lithium or carbamazepine in the first year, a crossover to the other drug in the second year, and the combination of both medications in the third year. Psychosensory scores from patients with bipolar disorder were compared with scores from healthy controls and with a variety of retrospective and prospective course of illness and treatment variables. Psychosensory symptoms occurred frequently in patients with bipolar I and II disorders, but were rare in healthy controls. When depressed, patients with bipolar II disorder (n = 23) reported more psychosensory symptoms when compared to patients with bipolar I disorder (n = 28), and those with a history of rapid cycling (n = 29) reported more psychosensory symptoms when compared to patients without a history of rapid cycling (n = 21). Psychosensory symptoms were not related to response to carbamazepine, lithium, or the combination of both drugs. Although the presence of psychosensory symptoms is associated with some bipolar subtypes (patients with bipolar II disorder and patients with a history of rapid cycling), they do not appear to predict treatment response. Further studies are needed to assess the pathophysiologic implications of the presence of psychosensory symptoms and their potential implications, if any, for directing therapeutics. PMID- 9359119 TI - Pattern of neuropsychologic dysfunction in inactive systemic lupus erythematosus. AB - The pattern of neuropsychological dysfunction in patients with inactive systemic lupus erythematosus (SLE) was examined. Fifty-eight subjects with inactive SLE and 47 healthy controls were administered a standardized neuropsychological test battery. Summary scores reflecting 18 different cognitive processes were derived. Subjects were designated cognitively impaired if three or more summary scores differed significantly from premorbid estimates of cognitive functioning. Cognitive impairment was identified in 43% of subjects with inactive SLE and 19% of healthy controls. Subjects with inactive SLE, as a group, performed significantly worse than healthy controls on measures of auditory verbal memory, visual spatial memory, psychomotor speed, and motor functioning. A significantly greater proportion of subjects with inactive SLE than healthy controls was impaired only on a measure of visual spatial memory. Cognitive impairment in subjects with inactive SLE was associated with increasing age. There were no associations between cognitive impairment and current depressive symptoms or current corticosteroid use. These findings suggest that cognitive dysfunction occurs frequently in inactive SLE. The variability of performance of subjects with inactive SLE is consistent with the heterogeneity of CNS involvement in the disease. PMID- 9359120 TI - Age differences in intention to left and right hemispace using a dichotic listening paradigm. AB - This study assessed the influence of age (younger women and elderly women living in communities) on cerebral laterality using dichotic listening. Previous research has purported to show a relative right cerebral decline with age. To date, however, research on the right hemiaging hypothesis has provided mixed findings. It is possible that these mixed findings are caused by use of simple versus complex dichotic listening tasks. As a test of this hypothesis, older women were expected to have a heightened right ear advantage (REA) for phonemic speech sounds and greater difficulty switching intention to the left ear when instructed to focus to either the left or the right ear. No age difference was found using the traditional presentation of concurrent phonemes. However, the right hemiaging hypothesis was supported on the intentional task, in which older women were less able to switch intention to the left but not to the right ear. Implications for right hemiaging are discussed. PMID- 9359121 TI - Frontal functions in juvenile myoclonic epilepsy. AB - The authors investigated cognition in juvenile myoclonic epilepsy (JME), focusing on frontal functions as suggested by maximal spatial distribution of epileptiform activity seen over frontocentral regions. Fifteen patients with JME (mean age, 34.3 years; mean estimated IQ 101) were administered a battery of tests sensitive to frontal dysfunction. The number of patients with impaired test performance and the frequency of impairment per test were calculated. Performance on selected tests was compared with that of 15 patients with temporal lobe epilepsy (TLE) who were matched for estimated IQ using paired t-tests. Although the performance of the group with JME was not uniform--some patients showed marked impairment whereas others showed little or no deficit--a high frequency of impairment was found on tests of concept formation-abstract reasoning and mental flexibility, cognitive speed, and planning and organization. Significant differences were found between the group with JME and the group with TLE on tests requiring mental flexibility and concept formation-abstract reasoning. In conjunction with studies demonstrating intractable seizures in approximately 20% of patients, the results from this study suggest that JME is not a uniformly benign condition. Frontal deficits may have maladaptive behavioral consequences suggestive of personality dysfunction, as described anecdotally by previous investigators. PMID- 9359122 TI - Consistency of within-day and across-day performance after mild brain injury. AB - The objective of this study was to determine whether inconsistent and erratic within-day and across-day performance is a symptom of mild to moderate traumatic brain injury (TBI), and to determine whether impaired consistency of performance can coexist, in the same patient, with intact or "normal" performance on single administrations of neuropsychological and other cognitive tests. The design was a matched-pair study in which a computerized cognitive test battery was administered 30 times over 4 days to all subjects. Performance patterns between TBI and control subjects were compared. Subjects also received traditional neuropsychological testing. The setting was a rehabilitation hospital outpatient department. The subjects were 12 adult volunteers, six with documented TBI and six with no history of TBI, neurologic illness, or injury. Control subjects showed consistent improvement of performance over days 1 to 4, whereas subjects with TBI showed erratic and inconsistent performance across days. In addition to inconsistent performance, some subjects with TBI showed worsening performance across days. The main outcome measures were performance on the Automated Neuropsychological Assessment Metrics (ANAM) battery and performance on traditional neuropsychological tests. Some patients with TBI in the study who have normal initial performance on traditional clinical neuropsychological tests and newly developed computerized cognitive tests show abnormalities of sustained performance. Such abnormalities are most apparent when performance is observed over multiple days, and are characterized by erratic and inconsistent across-day performance. Inconsistent performance was observed even in those subjects with TBI whose initial performance was equal to or better than that of the control subjects. Deficits in dynamic performance may explain why some patients with TBI who have excellent neuropsychological test performance nonetheless complain of functional decrement from premorbid ability. PMID- 9359123 TI - Akinetic mutism: disconnection of frontal-subcortical circuits. AB - Akinetic mutism may result from anterior cingulate lesions or a disconnection of the limbic connections projecting from the cingulate through subcortical circuits. Based on nonhuman primate primate tracer studies, ventral pallidal lesions should disrupt the anterior cingulate frontal-subcortical circuit. A patient developed a rigid akinetic mute state caused by bilateral lesions of the globus pallidus interna with ventral extension. The anatomic basis of the patient's clinical findings support a similarity in frontal-subcortical anatomy between humans and nonhuman primates. Isolated pallidal lesions are rare. Future studies should document whether ventral extension below the anterior commissure is associated with a loss of motivation. PMID- 9359124 TI - Benzodiazepine receptor uptake in a patient with panic disorder after citalopram treatment. PMID- 9359365 TI - Three neural groups in the femoral chordotonal organ of the cricket Gryllus bimaculatus: central projections and soma arrangement and displacement during joint flexion. AB - The arrangement of neuronal somata and their displacement during joint flexion together with the central projection of the pro- and metathoracic femoral chordotonal organs (FCOs) in the cricket were investigated. The FCO consists of the partially fused ventral and dorsal scoloparia in the proximal femur. The ventrally located neurones (the ventral group) form chain-like rows in which somata became sequentially smaller distally and project their axons ipsilaterally to the dorso-lateral regions, giving off abundant branches and terminating in the region between the dorsal intermediate tract and the ventral intermediate tract in the thoracic hemiganglion. The dorsal scoloparium, composed of small, simply aggregated neurones, projects exclusively to the medioventral association centre (mVAC), which is known to be an auditory neuropile. In addition, another neural cluster (the dorsal group) was found in the proximo-dorsal region of the ventral scoloparium. This was composed of simply aggregated neurones with axons giving off sparse branches dorso-laterally and terminating in the peripheral region inside the mVAC. The somata of these three groups were displaced distally by flexion of the femoro-tibial joint: the ventral group showed the greatest displacement, with the degree of movement depending upon soma location, while the dorsal group and dorsal scoloparium neurones were hardly displaced, possibly because of their strong connection with the cuticle. These properties were similar in both the prothoracic FCO and the metathoracic FCO. Taken together, the above points suggest that there is greater functional differentiation of the FCO than was previously thought. PMID- 9359366 TI - Sexual signalling in bladder grasshoppers: tactical design for maximizing calling range. AB - Pair formation in the bladder grasshopper (Bullacris membracioides) is by duetting and male phonotaxis. Low-frequency stridulatory signals are emitted by an abdominal resonator in the male and are answered by females using a species specific time delay. Acoustic transmission in the natural environment was studied using playback of sexual signals over distances of 450m under two atmospheric conditions (day and night). Upward-refracting sound conditions and a sound shadow zone beyond approximately 50m prevailed during the day. Acoustic enhancement was demonstrated at night when downward-refracting temperature inversions created a tunnel effect with sound caught between the ground and zones of different temperatures. Transmission conditions are almost ideal at night when the species actually calls; calling distances of 150m for the male signal in the afternoon increased to 1.5-1.9km at night, arguably the largest calling distance yet reported for insects. In contrast, female calls transmit over a maximum of 50m, signifying a marked discrepancy in the active space of sex-specific signals. Transmission distance may, however, be profoundly affected by levels of masking noise. Adaptations to increase the signal range may variously be found in the signal itself, in behaviour patterns or in the sensory system. Here we demonstrate aspects of the first two types of adaptation in the sexual signalling system of a grasshopper in which maximizing the calling range appears to be the major selection pressure, with lesser effects imposed by inter- and intraspecific pressures and by the transmission channel. PMID- 9359367 TI - Sulfide acquisition by the vent worm Riftia pachyptila appears to be via uptake of HS-, rather than H2S. AB - Deep-sea hydrothermal vents are home to a variety of invertebrate species, many of which host chemosynthetic bacteria in unusual symbiotic arrangements. The vent tubeworm Riftia pachyptila (Vestimentifera) relies upon internal chemolithoautotrophic bacterial symbionts to support its large size and high growth rates. Because of this, R. pachyptila must supply sulfide to the bacteria, which are far removed from the external medium. Internal H2S ([H2S+HS-+S2-]) can reach very high levels in R. pachyptila (2-12mmoll-1 in the vascular blood), most of which is bound to extracellular hemoglobins. The animal can potentially take up sulfide from the environment via H2S diffusion or via mediated uptake of HS-, or both. It was expected that H2S diffusion would be the primary sulfide acquisition mechanism, paralleling the previously demonstrated preferential uptake of CO2. Our data show, however, that the uptake of HS- is the primary mechanism used by R. pachyptila to obtain sulfide and that H2S diffusion into the worm apparently proceeds at a much slower rate than expected. This unusual mechanism may have evolved because HS- is less toxic than H2S and because HS- uptake decouples sulfide and inorganic carbon acquisition. The latter occurs via the diffusion of CO2 at very high rates due to the maintenance of an alkaline extracellular fluid pH. H2S accumulation is limited, however, to sulfide that can be bound by the hemoglobins, protecting the animal from sulfide toxicity and the symbionts from sulfide inhibition of carbon fixation. PMID- 9359368 TI - Why do tuna maintain elevated slow muscle temperatures? Power output of muscle isolated from endothermic and ectothermic fish. AB - It has been hypothesised that regional endothermy has evolved in the muscle of some tunas to enhance the locomotory performance of the fish by increasing muscle power output. Using the work loop technique, we have determined the relationship between cycle frequency and power output, over a range of temperatures, in isolated bundles of slow muscle fibres from the endothermic yellowfin tuna (Thunnus albacares) and its ectothermic relative the bonito (Sarda chiliensis). Power output in all preparations was highly temperature-dependent. A counter current heat exchanger which could maintain a 10 degrees C temperature differential would typically double maximum muscle power output and the frequency at which maximum power is generated (fopt). The deep slow muscle of the tuna was able to operate at higher temperatures than slow muscle from the bonito, but was more sensitive to temperature change than more superficially located slow fibres from both tuna and bonito. This suggests that it has undergone some evolutionary specialisation for operation at higher, but relatively stable, temperatures. fopt of slow muscle was higher than the tailbeat frequency of undisturbed cruising tuna and, together with the high intrinsic power output of the slow muscle mass, suggests that cruising fish have a substantial slow muscle power reserve. This reserve should be sufficient to power significantly higher sustainable swimming speeds, presumably at lower energetic cost than if intrinsically less efficient fast fibres were recruited. PMID- 9359369 TI - Ventilation and gas exchange in lizards during treadmill exercise. AB - The extent to which lizards ventilate their lungs during locomotion is controversial. Direct measurements of airflow across the nostrils suggest a progressive reduction in tidal volume and minute ventilation with increased running speed, while other studies have demonstrated that arterial PO2 remains constant during exercise. To resolve these conflicting findings, we measured minute ventilation and gas exchange rate in five specimens of Varanus exanthematicus and five specimens of Iguana iguana during treadmill locomotion at speeds between 0.14 and 1.11ms-1 at 35 degrees C. These speeds are much lower than maximal running speeds, but are greater than the maximal aerobic speed. In both species, the ventilatory pattern during locomotion was highly irregular, indicating an interference between locomotion and lung ventilation. In Varanus exanthematicus, treadmill locomotion elicited a six- to eightfold increase in minute ventilation from a pre-exercise level of 102mlkg-1min-1, whereas the rate of oxygen uptake increased approximately threefold (from 3.9 to 12.6mlkg-1min-1). After exercise, both minute ventilation and gas exchange rate decreased immediately. Because minute ventilation increased more than did oxygen consumption, an increase in lung PO2 during exercise is predicted and, thus, Varanus exanthematicus appears effectively to ventilate its lungs to match the increased metabolic rate during locomotion at moderate speed. In Iguana iguana, both minute ventilation and gas exchange rate increased above resting values during locomotion at 0.28ms-1, but both decreased with further increases in locomotor speed. Furthermore, following exercise, both minute ventilation and oxygen uptake rate increased significantly. Iguana iguana, therefore, appears to be unable to match the increased oxygen demand with adequate ventilation at moderate and higher speeds. PMID- 9359371 TI - Energetics of swimming by the platypus Ornithorhynchus anatinus: metabolic effort associated with rowing. AB - The metabolism of swimming in the platypus Ornithorhynchus anatinus Shaw was studied by measurement of oxygen consumption in a recirculating water flume. Platypuses swam against a constant water current of 0.45-1.0 ms-1. Animals used a rowing stroke and alternated bouts of surface and submerged swimming. Metabolic rate remained constant over the range of swimming speeds tested. The cost of transport decreased with increasing velocity to a minimum of 0.51 at 1.0 ms-1. Metabolic rate and cost of transport for the platypus were lower than values for semiaquatic mammals that swim at the water surface using a paddling mode. However, relative to transport costs for fish, the platypus utilized energy at a similar level to highly derived aquatic mammals that use submerged swimming modes. The efficient aquatic locomotion of the platypus results from its specialised rowing mode in conjunction with enlarged and flexible forefeet for high thrust generation and a behavioral strategy that reduces drag and energy cost by submerged swimming. PMID- 9359370 TI - 3-Hydroxybutyrate co-infused with noradrenaline decreases resulting plasma levels of noradrenaline in Wistar rats. AB - Pentobarbital-anaesthetized male Wistar rats were infused with 6microgkg-1min-1 of noradrenaline. The infusion was supplemented with 8.5 mgkg-1min-1 of D-3 hydroxybutyrate (3-OHB) for 15 min in order to determine its effect on the adrenergic response of the rat. Plasma levels of noradrenaline rose to a plateau of approximately 50 nmoll-1 with infusion. In the group infused with noradrenaline alone, noradrenaline levels were maintained for 1h. Supplementation with 3-OHB induced a decrease in plasma noradrenaline level that was inversely correlated with 3-OHB level. Aortic and interscapular brown adipose tissue temperatures increased with noradrenaline infusion, but the rise was arrested by 3-OHB; replacing 3-OHB with glucose had no effect. Infusion of saline, glucose or 3-OHB in the absence of noradrenaline did not induce a rise in temperature in either tissue. Blood 3-OHB concentration increased to 1.2 mmoll-1 during 3-OHB infusion, decreasing rapidly at the end of infusion. Blood glucose levels increased with noradrenaline infusion; the presence of high 3-OHB levels decreased glucose concentration. The effects observed were transient and dependent on 3-OHB concentration; these effects may help explain most of the other effects of noradrenaline described here. The role of 3-OHB as a regulator of adrenergic responses seems to be part of a complex fail-safe mechanism which prevents wasting. PMID- 9359372 TI - A physiological evaluation of carbon sources for calcification in the octocoral Leptogorgia virgulata (Lamarck). AB - The union of calcium cations with carbonate anions to form calcium carbonate (CaCO3) is a fundamentally important physiological process of many marine invertebrates, in particular the corals. In an effort to understand the sources and processes of carbon uptake and subsequent deposition as calcium carbonate, a series of studies of the incorporation of 14C-labeled compounds into spicules was undertaken using the soft coral Leptogorgia virgulata. It has been surmised for some time that dissolved inorganic carbon in sea water is used in the biomineralization process. Furthermore, it was suspected that metabolically generated CO2 is also available for calcification. As a means of testing these possible sources of carbon in spicule calcification, key enzymes or transport systems in each pathway were inhibited. First, the enzyme carbonic anhydrase was specifically inhibited using acetazolamide. Second, the active transport of bicarbonate was inhibited using DIDS (4,4'-diisothiocyanato-stilbene-2,2' disulfonic acid). Third, CO2 generation resulting from glycolysis and the citric acid cycle was arrested using iodoacetic acid, which interferes specifically with the enzyme glyceraldehyde-3-phosphate dehydrogenase. The results indicate that dissolved CO2 is the largest source of carbon used in the formation of calcitic sclerites, followed by HCO3- from dissolved inorganic carbon. In L. virgulata, the dissolved inorganic carbon is responsible for approximately 67% of the carbon in the sclerites. The other 33% comes from CO2 generated by glycolysis. Two important conclusions can be drawn from this work. First, carbon for spiculogenesis comes not only from dissolved inorganic carbon in the environment but also from metabolically produced carbon dioxide. While the latter has been theorized, it has never before been demonstrated in octocorals. Second, regardless of the carbon source, the enzyme carbonic anhydrase plays a pivotal role in the physiology of spicule formation in Leptogorgia virgulata. PMID- 9359373 TI - The effect of metamorphosis on the repeatability of maximal locomotor performance in the Pacific tree frog Hyla regilla. AB - Measuring the repeatability of inter-individual differences in locomotor performance is an important first step in elucidating both the functional causes and the ecological consequences of performance variation. Thus, repeatability of whole-animal performance traits provides a crucial link between functional and evolutionary biology. In the present study, repeatability of maximal burst locomotor performance was estimated for a single population of the Pacific tree frog Hyla regilla. Animals were reared individually from eggs through metamorphosis in the laboratory. Maximum burst swimming speed of tadpoles was measured before metamorphosis (Gosner stage 37) and again at the onset of the metamorphic climax (stage 42). Maximum jump distance was measured on the same individuals as juvenile frogs. Locomotor performance was repeatable over a 24h period for both premetamorphic tadpoles and juvenile frogs. Performance was not repeatable across metamorphosis or between any two of the three developmental stages investigated. A high-performance individual at one developmental stage does not necessarily retain that performance advantage at another stage. This lack of repeatability contrasts sharply with several previous studies on non metamorphosing vertebrates, but concurs with a single previous study on a metamorphosing salamander. Metamorphosis appears to place strict temporal constraints on individual consistency in locomotor ability. PMID- 9359374 TI - Cyclic AMP induces a relaxation response in the bullfrog Rana catesbeiana, but nitric oxide does not. AB - Cholecystokinin octapeptide (CCK), acetylcholine (ACh) and ceruletide have been shown to produce contraction in bullfrog (Rana catesbeiana) gallbladder strips. Agents capable of relaxing the bullfrog gallbladder are less numerous. Calcitonin gene-related peptide reduced the amount of both CCK- and ACh-induced tension in bullfrog gallbladder strips. The purpose of this study was to determine whether vasoactive intestinal peptide (VIP), nitric oxide (NO) and the second messengers cyclic GMP or cyclic AMP had any effect on gallbladder motility in the bullfrog. In vitro tension studies using l-NG-nitro-arginine methyl ester, Methylene Blue, sodium nitroprusside and N2,2'-O-dibutyryl guanosine 3',5'-cyclic monophosphate suggested that nitric oxide did not modulate gallbladder motility in the bullfrog gallbladder. Histochemical staining for NADPH diaphorase (nitric oxide synthase) failed to demonstrate nerve fibers containing nitric oxide synthase in the bullfrog gallbladder. In vitro studies demonstrated that VIP had no effect on CCK induced tension. However, in vitro studies using either 8-bromoadenosine 3',5' cyclic monophosphate or forskolin demonstrated that both agents relaxed strips precontracted with CCK. The results of this study suggested that, while neither NO nor VIP had a role in modulating bullfrog gallbladder motility, cyclic AMP was capable of modulating bullfrog gallbladder motility. PMID- 9359397 TI - Mechanism and regulation of Mg-chelatase. AB - Mg-chelatase catalyses the insertion of Mg into protoporphyrin IX (Proto). This seemingly simple reaction also is potentially one of the most interesting and crucial steps in the (bacterio)chlorophyll (Bchl/Chl)-synthesis pathway, owing to its position at the branch-point between haem and Bchl/Chl synthesis. Up until the level of Proto, haem and Bchl/Chl synthesis share a common pathway. However, at the point of metal-ion insertion there are two choices: Mg2+ insertion to make Bchl/Chl (catalysed by Mg-chelatase) or Fe2+ insertion to make haem (catalysed by ferrochelatase). Thus the relative activities of Mg-chelatase and ferrochelatase must be regulated with respect to the organism's requirements for these end products. How is this regulation achieved? For Mg-chelatase, the recent design of an in vitro assay combined with the identification of Bchl-biosynthetic enzyme genes has now made it possible to address this question. In all photosynthetic organisms studied to date, Mg-chelatase is a three-component enzyme, and in several species these proteins have been cloned and expressed in an active form. The reaction takes place in two steps, with an ATP-dependent activation followed by an ATP-dependent chelation step. The activation step may be the key to regulation, although variations in subunit levels during diurnal growth may also play a role in determining the flux through the Bchl/Chl and haem branches of the pathway. PMID- 9359398 TI - Troponin I and troponin T interact with troponin C to produce different Ca2+ dependent effects on actin-tropomyosin filament motility. AB - We have developed an in vitro motility assay to make a detailed quantitative analysis of Ca2+ control of skeletal-muscle troponin-tropomyosin control of actin filament movement over immobilized myosin. Ca2+ regulates both filament velocity and the fraction of filaments that are motile. We have demonstrated that the two effects are due to separate interactions of troponin C with troponin I and troponin T. When 64nM of the complex actin-tropomyosin-troponin I-troponin C was added at pCa 5, more than 80% of filaments were moving and their velocity did not change. At pCa 9, more than 20% of the filaments were moving. When 20nM of the complex actin-tropomyosin-troponin T+troponin I+troponin C was added at pCa 5, filament motility remained high, whereas velocity increased. The 30% increase in velocity observed when troponin T was present was also observed when heavy meromyosin fragment 1 labelled with N-ethylmaleimide (NEM S-1) was added after actin-tropomyosin filaments. The NEM S-1 effect was not additive with the troponin T-dependent velocity increase. The pattern of motile behaviour is characteristic of myosin on silicone-treated glass and different from the behaviour on nitrocellulose-coated glass. PMID- 9359400 TI - The purified and reconstituted ornithine/citrulline carrier from rat liver mitochondria: electrical nature and coupling of the exchange reaction with H+ translocation. AB - The mechanism and the electrical nature of ornithine/citrulline exchange has been investigated in proteoliposomes reconstituted with the ornithine/citrulline carrier purified from rat liver mitochondria. The stoichiometry of the exchanging substrates was close to 1:1. The exchange was not affected by inducing electrogenic flux of K+ with valinomycin. In contrast, the pH gradient generated by the K+/H+ exchanger nigericin in the presence of an outwardly directed K+ gradient stimulated the ornithineout/citrullinein exchange, but not the ornithine/ornithine homoexchange. Experiments in which either the internal or the external pH was varied, while keeping constant the pH in the other compartment, indicated that maximal exchange rates are found at pH 6 in the compartment containing citrulline and at pH 8 in the compartment containing ornithine. Changes in fluorescence of the pH indicator pyranine, included inside the proteoliposomes, showed that the exchanges ornithineout/citrullinein and citrullineout/ornithinein are accompanied by translocation of H+ in the same direction as citrulline. It is concluded that the mitochondrial ornithine/citrulline carrier catalyses an electroneutral exchange of ornithine+ for citrulline plus an H+. A reasonable model is one in which ornithine binds to a deprotonated carrier and citrulline to a protonated carrier and both substrate carrier complexes are neutral. The physiological implications of this transport process are discussed. PMID- 9359399 TI - The heat-shock transcription factor HSF1 is rapidly activated by either hyper- or hypo-osmotic stress in mammalian cells. AB - Osmoregulation, the cellular response to environmental changes of osmolarity and ionic strength, is important for the survival of living organisms. We have demonstrated previously that an exposure of mammalian cells to hypo-osmotic stress, either in growth medium (30% growth medium and 70% water) or in binary solution containing sorbitol and water, prominently induced the DNA-binding activity of the heat-shock transcription factor (HSF1) [Huang, Caruccio, Liu and Chen (1995) Biochem. J. 307, 347-352]. Since hyperosmotic and hypo-osmotic stress usually elicit opposite biological responses, we wondered what would be the effect of hyperosmotic stress on HSF activation. In this study we have examined the HSF DNA-binding activity in HeLa cells maintained in the sorbitol/water binary solution over a wide concentration range (0.1-0.9 M) and in Dulbecco's medium supplemented with sorbitol or NaCl. We found that HSF-binding activity could be induced prominently under both hypo-osmotic (0.1-0.25 M) and hyperosmotic conditions (0.50-0.90 M). In both cases, HSF activation was observed within 5 min after changing the osmotic pressure. The activation was accompanied by both HSF trimerization and nuclear translocation, and appeared to be independent of protein synthesis. The effects of hypo- or hyper-osmotic stress on HSF activation could be reversed once the cells were returned to iso-osmotic conditions (0.30M) with a half-life (t12) of 25 min or less. This rapid turnover of the osmotic-stress-induced HSF-binding activity was inhibited by cycloheximide, a potent inhibitor of protein synthesis. Unlike heat shock, activation of HSF by either hypo- or hyper-osmotic stress did not lead to an accumulation of heat-shock protein 70 (HSP70) mRNA in HeLa cells. We propose that HSF activation during osmotic stress may serve physiological functions independent of the synthesis of heat-shock proteins. PMID- 9359401 TI - Biochemical characterization of Caenorhabditis elegans UBC-1: self-association and auto-ubiquitination of a RAD6-like ubiquitin-conjugating enzyme in vitro. AB - The Caenorhabditis elegans ubiquitin-conjugating enzyme UBC-1 is distinct from other RAD6 homologues in possessing a C-terminal tail 40 amino acid residues long [Leggett, Jones and Candido (1995) DNA Cell Biol. 14, 883-891]. Such extensions from the core catalytic domain have been found in a subset of known conjugating enzymes, where they have been shown to have diverse roles including target recognition, membrane attachment and sporulation. In the present study we used mutagenesis in vitro to examine the role of the tail in specific aspects of UBC-1 structure and activity. Cross-linking experiments with purified recombinant UBC-1 reveal that it forms dimers and probably tetramers. The acidic tail of UBC-1 has an important role in this interaction because deletions of the tail significantly decrease, but do not abolish, this self-association. Ubiquitin conjugation assays show that, in addition to accepting a thiol-bound ubiquitin at its active site, UBC-1 is stably mono-ubiquitinated. Deletion analysis and site-directed mutagenesis localize the site of ubiquitination to Lys-162 in the tail. These findings demonstrate that the C-terminal tail of UBC-1 is important both for its quaternary structure and post-translational modification in vitro. PMID- 9359402 TI - Fatty acid signals in Bacillus megaterium are attenuated by cytochrome P-450 mediated hydroxylation. AB - In previous publications [English, Hughes and Wolf (1994) J. Biol. Chem. 269, 26836-26841; English, Hughes and Wolf (1996) Biochem. J. 316, 279-283], we have demonstrated that peroxisome proliferators and non-steroidal anti-inflammatory drugs are inducers of the cytochrome P-450BM-3 gene in Bacillus megaterium ATCC14581. Their mechanism of action involves binding to and subsequent displacement of the transcriptional repressor, Bm3R1, from its operator site, which results in the activation of cytochrome P-450BM-3 gene transcription. We now present evidence that the branched-chain fatty acid, phytanic acid, is a potent inducer of cytochrome P-450BM-3. We have also observed that phytanic acid and peroxisome proliferators are inducers of Bm3R1 protein accumulation and associated DNA-binding activity. In contrast, several barbiturates, although capable of inducing cytochrome P-450BM-3 and Bm3R1 gene transcription, were unable to induce the Bm3R1 protein. We also demonstrate that cytochrome P-450BM-3 readily oxidizes phytanic acid, and provide evidence that, although the omega-1 hydroxy acid derivatives of phytanic acid can associate with Bm3R1, they do so with an affinity two orders of magnitude lower than the unmodified fatty acid. As a consequence, the ability of the hydroxylated product to induce cytochrome P 450BM-3 gene expression in vivo is markedly reduced. These data collectively suggest that metabolism of fatty acids by cytochrome P-450BM-3 leads to an attenuation of their ability to activate the transcription of the BM-3 operon. This work places the action of bacterial fatty acid hydroxylases in an autoregulatory loop where they may be responsible for the inactivation or clearance of the inducing fatty acid signal. PMID- 9359404 TI - Site-directed mutagenesis of the Actinomadura R39 DD-peptidase. AB - The role of various residues in the conserved structural elements of the Actinomadura R39 penicillin-sensitive dd-peptidase has been studied by site directed mutagenesis. Replacement of Ser-298 of the 'SDN loop' by Ala or Gly significantly decreased the kcat/Km value for the peptide substrate, but only by a factor of 15 and had little effect on the other catalytic properties. Mutations of Asn-300 of the same loop and of Lys-410 of the KTG triad yielded very unstable proteins. However, the N300S mutant could be purified as a fusion protein with thioredoxin that exhibited decreased rates of acylation by the peptide substrate and various cephalosporins. Similar fusion proteins obtained with the N300A, K410H and K410N mutants were unstable and their catalytic and penicillin-binding properties were very strongly affected. In transpeptidation reactions, the presence of the acceptor influenced the kcat/Km values, which suggested a catalytic pathway more complex than a simple partition of the acyl-enzyme between hydrolysis and aminolysis. These results are compared with those obtained with two other penicillin-sensitive enzymes, the Streptomyces R61 dd-peptidase and Escherichia coli penicillin-binding protein (PBP) 5. PMID- 9359403 TI - Acute regulation of glucose transport in a monocyte-macrophage cell line: Glut-3 affinity for glucose is enhanced during the respiratory burst. AB - Activation of the respiratory burst imposes acute metabolic demands on phagocytic cells. These are met by mobilizing internal energy stores and by increasing the utilization of exogenous energy, including glucose in the circulation. To determine whether the increased glucose uptake that is known to be associated with the respiratory burst involves the regulation of glucose transporter molecules, the intrinsic transport properties of glucose transporters on the macrophage cell line RAW 264.7 were determined after activation with PMA, N formyl-methionine-leucine-phenylalanine (fMLP) and the cytokines granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin 3 (IL 3). Treatment with PMA resulted in a 2-fold increase in respiratory burst activity within 10 min; this was associated with a 30-50% increase in 2 deoxyglucose uptake and a 4-fold increase in transporter affinity for glucose. Similarly, fMLP, GM-CSF and IL-3 treatments stimulated 2-deoxyglucose uptake that was associated with a 3-4-fold increase in transporter affinity for glucose. To determine whether the changes observed in 2-deoxyglucose uptake in response to PMA, fMLP and growth factors were influenced by phosphorylation of the sugar, 3-O methylglucose, which is not phosphorylated, was used. Increased 3-O-methylglucose uptake and increased transporter affinity for glucose were also observed after PMA, fMLP and GM-CSF treatments. Whereas both fMLP and GM-CSF stimulated superoxide production, IL-3 failed to activate respiratory burst activity. The protein kinase inhibitors genistein and staurosporine inhibited the increase in 2 deoxyglucose uptake observed with fMLP and GM-CSF, and partly reversed the affinity increase towards that of untreated control cells. In contrast, the phosphatidylinositol 3-kinase inhibitor wortmannin had little effect on 2 deoxyglucose uptake in response to these activators. Western blotting with subtype-specific antisera showed that Glut-3 was the predominant transporter on RAW 264.7 cells. These studies demonstrate that acute regulation of glucose transporters occurs in response to activators that promote respiratory burst activity, and show that this regulation involves both tyrosine kinases and protein kinase C activity. PMID- 9359405 TI - Metabolism of exogenous S-adenosylmethionine in isolated rat hepatocyte suspensions: methylation of plasma-membrane phospholipids without intracellular uptake. AB - Administration of S-adenosylmethionine (AdoMet), the main biological methyl donor, has been shown to exert favourable effects on liver disorders in man and animal models. The mechanism of action of AdoMet has, however, remained elusive, mainly owing to controversies with respect to its capacity to enter intact liver cells. Incubation of isolated rat hepatocytes with 2 or 50 microM -methyl-14C AdoMet showed that it was utilized predominantly to methylate cellular phospholipids, forming mainly phosphatidylcholine, although less than 0.2% of labelled AdoMet was found inside the cells. The concentration of neither AdoMet nor S-adenosylhomocysteine (AdoHcy), its demethylation product, was significantly elevated inside the cells. A slight elevation of intracellular AdoMet was only recorded on incubation with concentrations of AdoMet above 200 microM. AdoHcy, which does not penetrate cells, inhibited phospholipid methylation from [methyl 14C]AdoMet but not from [methyl-14C]Met. Elevation of intracellular AdoHcy by adenosine dialdehyde, an inhibitor of AdoHcy hydrolase, inhibited phospholipid methylation from [methyl-14C]Met, but virtually not at all from [methyl 14C]AdoMet. Taken together, these data indicate that exogenous AdoMet does not penetrate hepatocytes significantly but is utilized for phospholipid methylation on the outer surface of the plasma membrane. PMID- 9359407 TI - Characterization of endoproteases from plant peroxisomes. AB - In this work, the characterization of endoprotease (EP) isoenzymes in peroxisomes is reported for the first time in cell organelles purified from pea leaves (Pisum sativum L.). A comparative analysis of the endo-proteolytic activity in peroxisomes purified from young (15-day-old) and senescent (50-day-old) leaves was carried out. Peroxisomes purified from senescent leaves showed a much higher endo-proteolytic activity than organelles from young plants. A 16 h incubation with exogenous substrates was the threshold time for the detection of a linear increase in the endo-proteolytic activity of peroxisomes from senescent leaves. Three EP isoenzymes (EP2, EP4 and EP5), having molecular masses of 88, 64 and 50 kDa respectively, were found in young plants by using SDS/polyacrylamide-gradient gels co-polymerized with gelatin. However, four additional isoenzymes (EP1, EP3, EP6 and EP7), with molecular masses of 220, 76, 46 and 34 kDa respectively, were detected in senescent plants. All the isoenzymes detected in peroxisomes from both young and senescent leaves were neutral proteases. By using different class specific inhibitors, the electrophoretically separated EP isoenzymes were characterized as three serine-proteinases (EP1, EP3 and EP4), two cysteine proteinases (EP2 and EP6) and a metallo-proteinase (EP7), and EP5 might be a metal-dependent serine-proteinase. Moreover, a peroxisomal polypeptide of 64 kDa was recognized by an antibody against a thiol-protease. The serine-proteinase isoenzymes (EP1, EP3 and EP4), which represent approx. 70% of the total EP activity of peroxisomes, showed a notable thermal stability, not being inhibited by incubation at 50 degrees C for 1 h. PMID- 9359406 TI - Phosphorylation of erythropoietin receptors in the endoplasmic reticulum by pervanadate-mediated inhibition of tyrosine phosphatases. AB - Erythropoietin (EPO) is the major hormone regulating the proliferation of erythroid precursors and their differentiation into erythrocytes. Ligand binding to the erythropoietin receptor (EPO-R), a member of the cytokine receptor family, triggers Tyr phosphorylation of the surface form of the receptor, presumably mediated by the Janus kinase (JAK) 2. To study whether non-surface EPO-R can be phosphorylated, Ba/F3 cells stably transfected with EPO-R were treated with pervanadate (PV), which is widely used as a potent tool to inhibit cellular protein Tyr phosphatases, thus resulting in enhanced Tyr phosphorylation of cellular proteins. PV treatment caused the EPO-R to undergo Tyr phosphorylation in a time-dependent and dose-dependent manner. PV-mediated Tyr phosphorylation of EPO-R occurred at several intracellular sites including the endoplasmic reticulum (ER), because both endoglycosidase H (endo H)-resistant EPO-R and the ER-retained EPO-R mutant (DeltaWS1 EPO-R) were Tyr phosphorylated in response to PV. Moreover, in metabolic labelling experiments, endo H-sensitive EPO-R was also phosphorylated. The phosphorylated fraction accounted for only 30-50% of the newly synthesized EPO-R, the fraction that normally exits from the ER. Tyr phosphorylation could not be detected on proteolytic fragments of the EPO-R, suggesting that this is a highly regulated process. Unlike the wild-type (wt) EPO R, which was phosphorylated both on EPO binding and after inhibition of Tyr phosphatases by PV treatment, an EPO-R mutant (W282R EPO-R) that does not activate JAK2 was phosphorylated after PV treatment but not by EPO binding. Both EPO-R and JAK2 were phosphorylated with similar kinetics by PV treatment, suggesting that JAK2, as well as protein Tyr kinases different from JAK2, might mediate PV-dependent EPO-R phosphorylation. Furthermore the Tyr-phosphorylated ER retained EPO-R mutant DeltaWS1 co-immunoprecipitated with JAK2 kinase, indicating that the EPO-R might interact with JAK2 while in the ER. PMID- 9359408 TI - Recombinant expression of rat glycine N-methyltransferase and evidence for contribution of N-terminal acetylation to co-operative binding of S adenosylmethionine. AB - An expression vector was constructed that produced rat glycine N methyltransferase in Escherichia coli. Recombinant glycine N-methyltransferase was purified to homogeneity by DEAE-cellulose and gel-filtration chromatography, with a yield of more than 80 mg of pure enzyme from a 1 litre culture. HPLC of tryptic peptides and analysis of isolated peptides showed that the recombinant enzyme was structurally identical with the liver enzyme except for the absence of N-terminal blocking. The alpha-amino group of rat glycine N-methyltransferase is blocked by acetylation [Ogawa, Konishi, Takata, Nakashima and Fujioka (1987) Eur. J. Biochem. 168, 141-151]. In contrast with the liver enzyme, which shows sigmoidal kinetics toward S-adenosylmethionine at all pH values tested [Ogawa and Fujioka (1982) J. Biol. Chem. 257, 3447-3452], the recombinant enzyme exhibited hyperbolic kinetics at low pH and sigmoidal rate behaviour at high pH. The Hill coefficient increased with increasing pH and a pKa of 8.11 was obtained in this transition. The values of Vmax and Km for glycine were not different between the two enzymes. These results suggest that elimination of the positive charge at the N-terminal end either by acetylation or deprotonation is required for co operative behaviour. PMID- 9359409 TI - An additional mechanism of ribosome-inactivating protein cytotoxicity: degradation of extrachromosomal DNA. AB - Inhibition of protein synthesis by cleavage of the N-glycosidic bond of a specific adenine of 28 S rRNA has been accepted as the mechanism by which plant ribosome-inactivating proteins (RIPs) cause cytotoxicity. The cytotoxic action of gelonin on Plasmodium falciparum malaria parasites appears to occur by a different mechanism. Parasite intoxication, which is manifested by mitochondrial dysfunction and lack of nucleic acid synthesis in the erythrocytic cycle following exposure to the toxin, is caused by the elimination of the parasite 6 kb extrachromosomal (mitochondrial) DNA. This is the first report which demonstrates that the DNA-damaging activities of RIPs observed in vitro can contribute to their cytotoxicity. PMID- 9359410 TI - Activator-protein-1 binding potentiates the hypoxia-induciblefactor-1-mediated hypoxia-induced transcriptional activation of vascular-endothelial growth factor expression in C6 glioma cells. AB - The endothelial cell-specific mitogen vascular-endothelial growth factor (VEGF) plays a key role in both physiological and pathological angiogenesis. The up regulation of VEGF expression in response to reduced oxygen tension occurs through transcriptional and post-transcriptional mechanisms. To investigate the molecular mechanisms of transcriptional activation by hypoxia (1% oxygen), fine mapping of a hypoxia-responsive region of the human VEGF promoter was carried out using luciferase reporter-gene constructs in C6 glioma cells. Here, we report that the binding site of hypoxia-inducible factor 1 (HIF1) is crucial for the hypoxic induction of VEGF gene expression. However, an enhancer subfragment containing the HIF1 binding site was not sufficient to confer full hypoxia responsiveness. Addition of upstream sequences restored the full sensitivity to hypoxia induction. This potentiating effect is due to activator protein 1 binding. The 'potentiating' sequences are unable to confer hypoxia responsiveness on their own. Our results strongly suggest that in C6 glioma cells a complex array of trans-acting factors facilitates full transcriptional induction of VEGF gene expression by hypoxia. PMID- 9359411 TI - Interleukin-6-dependent and -independent regulation of the human C-reactive protein gene. AB - We have investigated the function of different mediators of the regulation of the human C-reactive protein (hCRP) gene in transgenic mice. hCRP was induced by lipopolysaccharide and wounding in interleukin-6 (IL-6) +/+ mice, but not in IL-6 -/- mice. This finding suggested that IL-6 is necessary for the induction of hCRP. However, injection of IL-6 did not induce the hCRP gene. Thus, the induction of hCRP by IL-6 seems to require an additional cofactor. Therefore, we screened different cytokines for their activity in IL-6 +/+ and IL-6 -/- mice. Surprisingly, interleukin-1beta, as well as oncostatin M or leukaemia inhibitory factor, led to an induction of hCRP in both genetic backgrounds. These results indicate an IL-6-dependent and -independent regulation of hCRP. These hCRP transgenic mice therefore represent a novel model system for defining the cytokine network involved in the regulation of acute-phase genes during the course of inflammation. PMID- 9359412 TI - Membrane-binding properties of phospholipase C-beta1 and phospholipaseC-beta2: role of the C-terminus and effects of polyphosphoinositides, G-proteins and Ca2+. AB - We have studied the binding of two G-protein-regulated phospholipase C (PLC) enzymes, PLCs-beta1 and -beta2, to membrane surfaces using sucrose-loaded bilayer phospholipid vesicles of varying compositions. Neither enzyme binds appreciably to pure phosphatidylcholine vesicles at lipid concentrations up to 10(-3) M. PLC beta1 and PLC-beta2 bind vesicles composed of phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine (molar ratio 1:1:1) with an approximate Kd of 10(-5) M. Inclusion of 2% PtdIns(4,5)P2 in these vesicles had no effect on the affinity of this interaction. As reported by others, removal of the C-terminus of PLC-beta1 and PLC-beta2 produces catalytically active fragments. The affinity of these truncated proteins for phospholipid vesicles is dramatically reduced suggesting that this region of the proteins contains residues important for membrane binding. Inclusion of G-protein alpha- and betagamma-subunit activators in the phospholipid vesicles does not increase the binding of PLC-beta1 or PLC-beta2, and the magnitude of G-protein-mediated PLC activation observed at low phospholipid concentrations (10(-6) M) is comparable to that observed at concentrations at which the enzymes are predominantly membrane-bound (10(-3) M). PLC-beta1 and -beta2 contain C2 domains but Ca2+ does not enhance binding to the vesicles. Our results indicate that binding of these enzymes to membranes involves the C-temini of the proteins and suggest that activation of these enzymes by G-proteins results from a regulated interaction between the membrane-bound proteins rather than G-protein-dependent recruitment of soluble enzymes to a substrate-containing phospholipid surface. PMID- 9359414 TI - Modification of the mitochondrial F1-ATPase epsilon subunit, enhancement of the ATPase activity of the IF1-F1 complex and IF1-binding dependence of the conformation of the epsilon subunit. AB - Treatment of bovine heart submitochondrial particles with a low concentration of 2-hydroxy-5-nitrobenzyl bromide (HNB), a selective reagent for the Trp residue of the epsilon subunit [Baracca, Barogi, Lenaz and Solaini (1993) Int. J. Biochem. 25, 1269-1275], enhances the ATP hydrolytic activity of the particles exclusively when the natural inhibitor protein IF1 is present. Similarly, isolated F1 [the catalytic sector of the mitochondrial H+-ATPase complex (ATP synthase)] treated with the reagent has the ATPase activity enhanced exclusively if IF1 is bound to it. These experiments suggest that the modification of the epsilon subunit decreases the inhibitory activity of IF1, eliciting the search for a relationship between the epsilon subunit and the inhibitory protein. Certainly, a reverse relationship exists because HNB binds covalently to the isolated F1 exclusively when the inhibitory protein is present. This finding is consistent with the existence of the epsilon subunit in different conformational states depending on whether IF1 is bound to F1 or not. Support for this assertion is obtained by measurements of the intrinsic phosphorescence decay rate of F1, a probe of the Trp epsilon subunit conformation in situ [Solaini, Baracca, Parenti-Castelli and Strambini (1993) Eur. J. Biochem. 214, 729-734]. A significant difference in phosphorescence decay rate is detected when IF1 is added to preparations of F1 previously devoid of the inhibitory protein. These studies indicate that IF1 and the epsilon subunit of the mitochondrial F1-ATPase complex are related, suggesting a possible role of the epsilon subunit in the mechanism of regulation of the mitochondrial ATP synthase. PMID- 9359413 TI - Lipoprotein lipase activity in rat cardiomyocytes is stimulated by insulin and dexamethasone. AB - Lipoprotein lipase (LPL) activity was studied in rat cardiomyocytes after overnight culture (16 h) in the presence of insulin (100 nM) and/or dexamethasone (100 nM). Insulin in combination with dexamethasone (INS/DEX) increased heparin releasable LPL activity by 71% over the control level (566+/-85 versus 331+/-48 nmol/h.mg cell protein). This was accompanied by a 61% increase in total cellular LPL activity (914+/-89 versus 567+/-64 nmol/h.mg cell protein). The increase in LPL activity occurred at sub-nanomolar concentrations of the hormones, but neither hormone was effective alone. LPL protein mass, quantified by ELISA, was the same in both control and INS/DEX-treated cells (27.7 versus 28.6 ng/mg cell protein, respectively), thus LPL specific activity in cardiomyocytes was increased by INS/DEX treatment (0.113 versus 0.069 mU/ng LPL protein). These findings emphasize the importance of hormonal interactions in the regulation of LPL in heart tissue. PMID- 9359415 TI - Gender difference in regulation of branched-chain amino acid catabolism. AB - Regulation of the activity state of the hepatic branched-chain 2-oxo acid dehydrogenase (BCODH) complex during the light-dark cycle differs markedly in male and female rats. Female rats exhibit a profound diurnal rhythm in the activity state of the complex that is not observed in male rats. Regardless of gender, most of the complex was dephosphorylated and active in the middle of the dark period and early in the light period, and this form of the complex predominated in male rats at the end of the light period. In contrast, most of the complex in female rats became phosphorylated and inactive by the end of the light period. Gonadectomy prevented the diurnal rhythm in females but was without effect in males, indicating that female sex hormones are required for this gender difference in regulation of the BCODH complex. Changes in levels of branched chain 2-oxo acids, known regulators of BCODH kinase, do not seem to be involved; rather, an increase in BCODH kinase activity occurring between morning and evening is responsible for inactivation of the BCODH complex in female rats. The increase in kinase activity is due to an increase in the amount of kinase protein associated with the BCODH complex. Thus a marked diurnal variation in the amount of BCODH kinase and therefore its activity results in large swings in the activity state of the liver BCODH complex in female rats. This study provides the first evidence for a gender-specific difference in the regulation of branched chain amino acid catabolism. PMID- 9359416 TI - Bovine retinal pigment epithelium contains novel types of phospholipase A2. AB - We have recently demonstrated the presence of phospholipase A2 (PLA2) activity in cells from bovine retinal pigment epithelium (RPE) [Jacob et al. (1996) J. Biol. Chem. 271, 19209-19218]. We report here our results on the characterization of this RPE-PLA2 activity. We show that RPE probably contains two types of PLA2 enzyme, as indicated by the results obtained with different PLA2-active fractions eluted from cation-exchange columns and treated with Ca2+/EGTA, dithiothreitol, p bromophenacyl bromide or heat. These results, in addition to those from PLA2 assays using different substrates, also suggest that RPE-PLA2 enzymes are different from the well-known secretory, cytoplasmic and Ca2+-independent forms. Sequential extraction of RPE with (1) isotonic, (2) hypertonic and (3) detergent containing PBS argues for the presence of weakly membrane-associated enzymes. Control experiments using 'back and forth' TLC allowed us to discriminate between PLA2 and phospholipase C/diacylglycerol lipase activity and confirmed that, in our assay conditions, the release of fatty acids was indeed due to PLA2 enzymes. These results, together with those obtained by treating RPE homogenates with H2SO4, guanosine 5'-[gamma-thio]triphosphate, ATP and different protease inhibitors, permitted us to make the first characterization of these RPE-PLA2 enzymes. We conclude that RPE contains novel types of PLA2 that are different from the secretory, cytoplasmic and Ca2+-independent forms. PMID- 9359418 TI - Proteoglycan synthesis in haematopoietic cells: isolation and characterization of heparan sulphate proteoglycans expressed by the bone-marrow stromal cell line MS 5. AB - Proteoglycans of bone-marrow stromal cells and their extracellular matrix are important components of the haematopoietic microenvironment. Recently, several studies have indicated that they are involved in the interaction of haematopoietic stem and stromal cells. However, a detailed characterization of the heparan sulphate proteoglycans synthesized by bone-marrow stromal cells is still lacking. Here we report on the isolation and characterization of proteoglycans from the haematopoietic stromal cell line MS-5, that efficiently supports the growth and differentiation of human and murine haematopoietic progenitor cells. Biochemical characterization of purified proteoglycans revealed that the haematopoietic stromal cell line MS-5 synthesizes, in addition to chondroitin sulphate proteoglycans, several different heparan sulphate proteoglycans. Immunochemical analysis, using specific antibodies against the different members of the syndecan family, glypican, betaglycan and perlecan, showed that MS-5 cells synthesize all these different heparan sulphate proteoglycans. These data were further supported by reverse-transcriptase PCR and confirmed by sequence and Northern blot analysis. The relative abundance of the different heparan sulphate proteoglycans was estimated on the protein and mRNA levels. PMID- 9359417 TI - Cholecystokinin-stimulated tyrosine phosphorylation of p125FAK and paxillin is mediated by phospholipase C-dependent and -independent mechanisms and requires the integrity of the actin cytoskeleton and participation of p21rho. AB - Recent studies show that the effects of some oncogenes, integrins, growth factors and neuropeptides are mediated by tyrosine phosphorylation of the cytosolic kinase p125 focal adhesion kinase (p125(FAK)) and the cytoskeletal protein paxillin. Recently we demonstrated that cholecystokinin (CCK) C-terminal octapeptide (CCK-8) causes tyrosine phosphorylation of p125(FAK) and paxillin in rat pancreatic acini. The present study was aimed at examining whether protein kinase C (PKC) activation, calcium mobilization, cytoskeletal organization and small G-protein p21(rho) activation play a role in mediating the stimulation of tyrosine phosphorylation by CCK-8 in acini. CCK-8-stimulated phosphorylation of p125(FAK) and paxillin reached a maximum within 2.5 min. The CCK-8 dose response for causing changes in the cytosolic calcium concentration ([Ca2+]i) was similar to that for p125(FAK) and paxillin phosphorylation, and both were to the left of that for receptor occupation and inositol phosphate production. PMA increased tyrosine phosphorylation of both proteins. The calcium ionophore A23187 caused only 25% of the maximal stimulation caused by CCK-8. GF109203X, a PKC inhibitor, completely inhibited phosphorylation with PMA but had no effect on the response to CCK-8. Depletion of [Ca2+]i by thapsigargin had no effect on CCK-8-stimulated phosphorylation. Pretreatment with both GF109203X and thapsigargin decreased CCK 8-stimulated phosphorylation of both proteins by 50%. Cytochalasin D, but not colchicine, completely inhibited CCK-8- and PMA-induced p125(FAK) and paxillin phosphorylation. Treatment with Clostridium botulinum C3 transferase, which inactivates p21(rho), caused significant inhibition of CCK-8-stimulated p125(FAK) and paxillin phosphorylation. These results demonstrate that, in pancreatic acini, CCK-8 causes rapid p125(FAK) and paxillin phosphorylation that is mediated by both phospholipase C-dependent and -independent mechanisms. For this tyrosine phosphorylation to occur, the integrity of the actin, but not the microtubule, cytoskeleton is essential as well as the activation of p21(rho). PMID- 9359419 TI - Carboxymethylation of nuclear protein serine/threonine phosphatase X. AB - Specific rabbit polyclonal antibodies against peptides corresponding to the highly homologous protein serine/threonine phosphatase 2A and X catalytic subunits (PP2A/C and PPX/C respectively) were used to investigate the cellular and subcellular distribution of PP2A/C and PPX/C, as well as their methylation state. Immunoblots of rat tissue extracts revealed a widespread distribution of these enzymes but particularly high levels of PP2A/C and PPX/C in brain and testes respectively. In addition, immunoblots of subcellular fractions and immunocytochemical analyses of rat brain sections demonstrated that PPX/C is predominantly localized to the nucleus, whereas PP2A/C is largely cytoplasmic. Treatment of nuclear extracts with alkali resulted in increased PPX/C immunoreactivity to a polyclonal antibody directed against the C-terminus; no change in PPX immunoreactivity was observed using an antibody against an internal peptide. Alkali treatment of brain and liver cytosolic and nuclear extracts did not change the molecular mass or the isoelectric point of PPX/C. Furthermore, tritiated PPX/C was immunoprecipitated from COS cell extracts incubated with the methyl donor S-adenosyl-l-[methyl-3H]methionine. Thus the increase in immunoreactivity probably results from removal of a carboxymethyl group from PPX/C, as has been shown previously for PP2A/C [Favre, Zolnierowicz, Turowski and Hemmings (1994) J. Biol. Chem. 269, 16311-16317]. Together, our results indicate that the PPX catalytic subunit is a predominantly nuclear phosphatase and is methylated at its C-terminus. PMID- 9359421 TI - Transient times in linear metabolic pathways under constant affinity constraints. AB - In the early seventies, Easterby began the analytical study of transition times for linear reaction schemes [Easterby (1973) Biochim. Biophys. Acta 293, 552 558]. In this pioneer work and in subsequent papers, a state function (the transient time) was used to measure the period before the stationary state, for systems constrained to work under both constant and variable input flux, was reached. Despite the undoubted usefulness of this quantity to describe the time dependent features of these kinds of systems, its application to the study of chemical reactions under other constraints is questionable. In the present work, a generalization of these magnitudes to linear metabolic pathways functioning under a constant-affinity constraint is carried out. It is proved that classical definitions of transient times do not reflect the actual properties of the transition to the steady state in systems evolving under this restriction. Alternatively, a more adequate framework for interpretation of the transient times for systems with both constant and variable input flux is suggested. Within this context, new definitions that reflect more accurately the transient characteristics of constant affinity systems are stated. Finally, the meaning of these transient times is discussed. PMID- 9359420 TI - Thiocyanate and chloride as competing substrates for myeloperoxidase. AB - The neutrophil enzyme myeloperoxidase uses H2O2 to oxidize chloride, bromide, iodide and thiocyanate to their respective hypohalous acids. Chloride is considered to be the physiological substrate. However, a detailed kinetic study of its substrate preference has not been undertaken. Our aim was to establish whether myeloperoxidase oxidizes thiocyanate in the presence of chloride at physiological concentrations of these substrates. We determined this by measuring the rate of H2O2 loss in reactions catalysed by the enzyme at various concentrations of each substrate. The relative specificity constants for chloride, bromide and thiocyanate were 1:60:730 respectively, indicating that thiocyanate is by far the most favoured substrate for myeloperoxidase. In the presence of 100 mM chloride, myeloperoxidase catalysed the production of hypothiocyanite at concentrations of thiocyanate as low as 25 microM. With 100 microM thiocyanate, about 50% of the H2O2 present was converted into hypothiocyanite, and the rate of hypohalous acid production equalled the sum of the individual rates obtained when each of these anions was present alone. The rate of H2O2 loss catalysed by myeloperoxidase in the presence of 100 mM chloride doubled when 100 microM thiocyanate was added, and was maximal with 1mM thiocyanate. This indicates that at plasma concentrations of thiocyanate and chloride, myeloperoxidase is far from saturated. We conclude that thiocyanate is a major physiological substrate of myeloperoxidase, regardless of where the enzyme acts. As a consequence, more consideration should be given to the oxidation products of thiocyanate and to the role they play in host defence and inflammation. PMID- 9359422 TI - Chondroitin sulphate composition and structure in alternatively spliced CD44 fusion proteins. AB - Previous studies have indicated that CD44 isoforms, spliced with variant exons, are heterogeneously glycanated with chondroitin sulphate and heparan sulphate chains. Because such alternative splicing may regulate divergent biological effects of the specific isoforms, we analysed the consequences of this process on the composition and structure of the chondroitin-sulphate chains. Recombinant chimaeras were engineered with and without exons V3-10 or V3,8-10 and expressed as Ig fusion proteins in COS cells. In addition, the chondroitin sulphates of wild-type isoforms were contrasted with those of isoforms mutated with serine-to alanine codon substitutions at a putative Ser-Gly-Ser-Gly glycosaminoglycan acceptor site within exon V3. The chondroitin sulphates contained both 4- and 6 sulphated galactosamine residues, although there was a high content of non sulphated galactosamine-containing repeat units. Splicing of exons V4-7, which contain no Ser-Gly consensus motifs, resulted in increased glycanation with chondroitin-sulphate chains, as well as increased sulphation levels of the polymers. Comparison of wild-type and acceptor-site mutant isoforms showed that chondroitin-sulphate content declined by more than 60-80% in the mutant, indicating that assembly of chondroitin-sulphate chains occurs there, and a general decrease in the sulphation level of the remaining chains was observed. Undersulphation of the recombinant chondroitin sulphates was shown by parallel analyses with native human keratinocyte CD44 molecules and is most probably an artifact of transient expression in COS cells. Our data indicate that combinatorial exon splicing exerts complex and distal effects on glycanation patterns and structure, which presumably modulate those functions that may be mediated though the chondroitin-sulphate moieties, such as motility and matrix invasion. PMID- 9359423 TI - Multiple muscle-specific regulatory elements are associated with a DNase I hypersensitive site of the cardiac beta-myosin heavy-chain gene. AB - Using nuclei isolated from neonatal cardiomyocytes, we have mapped the DNase I hypersensitive sites (DHSs) residing within the 5'-upstream regions of the hamster cardiac myosin heavy-chain (MyHC) gene. Two cardiac-specific DHSs within the 5 kb upstream region of the cardiac MyHC gene were identified. One of the DHSs was mapped to the -2.3 kb (beta-2.3 kb) region and the other to the proximal promoter region. We further localized the beta-2.3 kb site to a range of 250 bp. Multiple, conserved, muscle regulatory motifs were found within the beta-2.3 kb site, consisting of three E-boxes, one AP-2 site, one CArG motif, one CT/ACCC box and one myocyte-specific enhancer factor-2 site. This cluster of regulatory elements is strikingly similar to a cluster found in the enhancer of the mouse muscle creatine kinase gene (-1256 to -1050). The specific interaction of the motifs within the beta-2.3 kb site and the cardiac nuclear proteins was demonstrated using gel mobility-shift assays and footprinting analysis. In addition, transfection analysis revealed a significant increase in chloramphenicol acetyltransferase activity when the beta-2.3 kb site was linked to a heterologous promoter. These results suggest that previously undefined regulatory elements of the beta-MyHC gene may be associated with the beta-2.3 kb site. PMID- 9359424 TI - Evidence for redox regulation of the transcription factor NtcA, acting both as an activator and a repressor, in the cyanobacterium Anabaena PCC 7120. AB - NtcA has been identified as a nitrogen-responsive regulatory protein required for nitrogen assimilation and heterocyst differentiation in cyanobacteria. It is proposed that NtcA functions through the formation of DNA-protein complexes with its specific target sequence within the promoter regions of the regulated genes. In vitro, NtcA of Anabaena PCC 7120 binds to upstream regions of the genes whose products are involved in nitrogen assimilation, but also to the upstream region of rbcLS (carbon-fixation gene), xisA (encoding a site-specific recombinase expressed during heterocyst differentiation) and ntcA (encoding NtcA itself). However, the mechanism by which NtcA serves as a critical regulator for such diverse processes is not understood. With the use of electrophoretic mobility shift assays, NtcA from Anabaena PCC 7120 was here shown to interact with the promoter sequence of the gor gene, encoding glutathione reductase, thereby providing a novel example of NtcA's acting as a repressor, previously found only for the rbcLS gene. Furthermore we demonstrate that the binding of DNA by NtcA is regulated in vitro by a redox-dependent mechanism involving cysteine residues of the NtcA protein. These findings suggest that NtcA is a transcriptional regulator that responds not only to the nitrogen status but also to the cellular redox status, a function that might be particularly significant during heterocyst differentiation. PMID- 9359425 TI - Galactosamine in walls of slow-growing mycobacteria. AB - Galactosamine was found consistently as a minor component of the envelope of five species of slow-growing mycobacteria, including all the major human pathogens, but not three rapid-growing species. The amino sugar was a component of the arabinogalactan of the cell wall skeleton, and occurred at the level of about one residue per arabinogalactan chain. Its amino group was in the free, un-N acetylated state. Examination of oligosaccharides released by partial acid hydrolysis of arabinogalactan by fast atom bombardment-MS and gas chromatography MS identified a series of oligoarabinans, each possessing one GalN unit, linked to position 2 of arabinose. It is proposed that the GalN residues occur as stub branches of 1-->5-linked arabinose chains in the arabinogalactan. Possible functions of GalN are discussed. PMID- 9359426 TI - Association of bile-salt-dependent lipase with membranes of human pancreatic microsomes is under the control of ATP and phosphorylation. AB - Bile-salt-dependent lipase (BSDL) is secreted by the pancreas into the duodenum, where it catalyses the hydrolysis of dietary lipid esters on activation by bile salts. The secretion pathway of BSDL is comparable with that of other digestive enzymes produced by pancreatic acinar cells. However, in contrast with these other enzymes, BSDL is partly associated with endoplasmic reticulum membranes as part of a folding complex, including a Grp94-related protein to which BSDL is transiently linked. The release of BSDL from membranes occurs once its glycosylation is completed [Bruneau and Lombardo (1995) J. Biol. Chem. 270, 13524 13533]. In the present study, investigations concerning the mechanism of association/dissociation of BSDL with membranes of microsomes were performed. For this purpose the role of ATP and that of the possible phosphorylation of BSDL were examined. For the first time, it is shown that human pancreatic BSDL is phosphorylated, probably at a serine residue, during its transport within the acinar cell. The phosphorylation of BSDL is provoked by calphostin C, an inhibitor of protein kinase C. In the presence of 1-(isoquinolinesulphonyl)2 methylpiperazine, a non-specific inhibitor of serine/threonine protein kinase A, C or G, or of calcium chelator 1,2-bis(O-aminophenoxy)ethane-N,N,N', N'-tetra acetic tetra(acetoxymethyl)ester, the phosphorylation of BSDL elicited by calphostin C is abolished. These data suggested that the phosphorylation of BSDL within human pancreatic microsomes is under the control of a cascade of protein kinases. We have also shown that the phosphorylation of BSDL appears to be involved in the release of the enzyme from microsome membranes. Nevertheless ATP, which modifies the conformation of BSDL, triggers this association, and an unhydrolysable ATP analogue was unable to promote it. PMID- 9359427 TI - Engineering the substrate specificity of Bacillus megaterium cytochrome P-450 BM3: hydroxylation of alkyl trimethylammonium compounds. AB - Oligonucleotide-directed mutagenesis has been used to replace arginine-47 with glutamate in cytochrome P-450 BM3 from Bacillus megaterium and in its haem domain. The mutant has been characterized by sequencing, mass spectrometry, steady-state kinetics and by optical and NMR measurements of substrate binding. The mutant retains significant catalytic activity towards C12-C16 fatty acids, catalysing hydroxylation in the same (omega-1, omega-2, omega-3) positions with kcat/Km values a factor of 14-21 lower. C12-C16 alkyl trimethylammonium compounds are relatively poor substrates for the wild-type enzyme, but are efficiently hydroxylated by the arginine-47-->glutamate mutant at the omega-1, omega-2 and omega-3 positions, with kcat values of up to 19 s-1. Optical spectroscopy shows that the binding of the C14 and C16 alkyl trimethylammonium compounds to the mutant is similar to that of the corresponding fatty acids to the wild-type enzyme. Paramagnetic relaxation measurements show that laurate binds to the ferric state of the mutant in a significantly different position, 1.5 A closer to the iron, than seen in the wild-type, although this difference is much smaller ( approximately 0.2 A) in the ferrous state of the complex. The binding of a substrate having the same charge as residue 47 to the ferric state of the enzyme is roughly ten times weaker than that of a substrate having the opposite charge (and thus is able to make an ion-pair interaction with this residue). The results are discussed in the light of the three-dimensional structure of the enzyme. PMID- 9359429 TI - Biological variability in the structures of diphosphoinositol polyphosphates in Dictyostelium discoideum and mammalian cells. AB - Previous structural analyses of diphosphoinositol polyphosphates in biological systems have relied largely on NMR analysis. For example, in Dictyostelium discoideum, diphosphoinositol pentakisphosphate was determined by NMR to be 4- and/or 6-PPInsP5, and the bisdiphosphoinositol tetrakisphosphate was found to be 4, 5-bisPPInsP4 and/or 5,6-bisPPInsP4 [Laussmann, Eujen, Weisshuhn, Thiel and Vogel (1996) Biochem. J. 315, 715-720]. We now describe three recent technical developments to aid the analysis of these compounds, not just in Dictyostelium, but also in a wider range of biological systems: (i) improved resolution and sensitivity of detection of PPInsP5 isomers by microbore metal-dye-detection HPLC; (ii) the use of the enantiomerically specific properties of a rat hepatic diphosphatase; (iii) chemical synthesis of enantiomerically pure reference standards of all six possible PPInsP5 isomers. Thus we now demonstrate that the major PPInsP5 isomer in Dictyostelium is 6-PPInsP5. Similar findings obtained using the same synthetic standards have been published [Laussmann, Reddy, Reddy, Falck and Vogel (1997) Biochem. J. 322, 31-33]. In addition, we show that 10-25% of the Dictyostelium PPInsP5 pool is comprised of 5-PPInsP5. The biological significance of this new observation was reinforced by our demonstration that 5 PPInsP5 is the predominant PPInsP5 isomer in four different mammalian cell lines (FTC human thyroid cancer cells, Swiss 3T3 fibroblasts, Jurkat T-cells and Chinese hamster ovary cells). The fact that the cellular spectrum of diphosphoinositol polyphosphates varies across phylogenetic boundaries underscores the value of our technological developments for future determinations of the structures of this class of compounds in other systems. PMID- 9359428 TI - Regulation of inositol lipid-specific phospholipase cdelta by changes in Ca2+ ion concentrations. AB - Studies of inositol lipid-specific phospholipase C (PLC) have elucidated the main regulatory pathways for PLCbeta and PLCgamma but the regulation of PLCdelta isoenzymes still remains obscure. Here we demonstrate that an increase in Ca2+ ion concentration within the physiological range (0.1-10 microM) is sufficient to stimulate PLCdelta1, but not PLCgamma1 and PLCbeta1, to hydrolyse cellular inositol lipids present in permeabilized cells. The activity of PLCdelta1 is further enhanced in the presence of phosphatidylinositol transfer protein (PI TP). Both full activation by Ca2+ ions and stimulation in the presence of PI-TP require an intact PH domain involved in the membrane attachment of PLCdelta1. The physiological implication of this study is that PLCdelta1 could correspond to a previously uncharacterized PLC responsible for Ca2+ ion-stimulated inositol lipid hydrolysis observed in many cellular systems. PMID- 9359430 TI - Functional conformations of the nuclear 1alpha,25-dihydroxyvitamin D3 receptor. AB - The nuclear hormone 1alpha,25-dihydroxyvitamin D3 (VD) has important cell regulatory functions. Various synthetic VD analogues are under investigation to identify candidates with an improved therapeutic profile against hyperproliferative diseases. VD directly activates the transcription factor VD receptor (VDR), which in turn stimulates the expression of a cascade of primary and secondary VD-responsive genes. The activation of the VDR through binding of its natural and synthetic ligands is linked to a conformational change presenting the interface with co-activator proteins, referred to as the (trans)activation function 2 (AF-2) domain. Multiple conformations of the VDR might be the key to understanding a selective action of VD analogues. The method of limited protease digestion was used here to characterize up to three different functional VDR conformations stabilized individually by VD and its analogues. The relative potency of VDR ligands can be quantified in the interaction with these VDR conformations by determination of a functional dissociation constant, where a two concentration-point comparison has already provided important information. In this way seven amino acid residues in the AF-2 domain have been analysed as potential ligand contact points. Interestingly, residues Phe-422 and Val-418 seem to interact with all tested VDR ligands, whereas VD analogues such as the anti psoriatic drug MC903 displayed additional contact points within the AF-2 domain. Taken together, limited protease digestion is a powerful method for studying functional VDR conformations and seems to be very appropriate for screening VD analogues. PMID- 9359431 TI - Metabolism and possible compartmentalization of inositol lipids in isolated rat liver nuclei. AB - (1) The removal of the nuclear envelope from isolated rat-liver nuclei by washing with Triton X-100 (TX-100) was assessed by electron microscopy. All the envelope was removed by 0.04% (w/v) TX-100. (2) After this removal, phosphorylation of inositol lipids and diacylglycerol (DAG) from [gamma-32P]ATP still occurs, despite the near complete absence of detectable (by mass assay) DAG and PtdIns. This suggests that the majority of these two lipids in nuclei are present in the nuclear membrane, but the small amounts remaining after extraction, defined as intranuclear, are available for phosphorylation by lipid kinases (36% for DAG and 24% for PtdIns respectively, when expressed as a percentage of incorporation of intact nuclei). (3) PtdIns(4,5)P2 did not follow the same pattern as PtdIns and DAG; after removal of the nuclear membrane, 40% of the mass of this lipid was left in the nucleus. Moreover, a similar amount of PtdIns(4,5)P2 was also resistant to extraction with even higher concentrations of detergent, suggesting that PtdIns(4,5)P2 has a discrete intranuclear location, probably bound to nuclear proteins. (4) Addition of exogenous substrates, PtdIns, PtdIns(4)P and DAG, to membrane-depleted nuclei resulted in reconstitution of the majority of lipid phosphorylations from [gamma-32P]ATP (70%, 90% and 94% of intact nuclei respectively), suggesting a predominantly intranuclear location for the respective kinases. (5) Nuclei also showed phosphomonoesterase and phosphatidic acid hydrolase activity; dephosphorylation of pre-radiolabelled PtdIns(4)P, PtdIns(4,5)P2 and phosphatidic acid was observed when [gamma-32P]ATP was removed. However, some of the radioactivity was apparently resistant to these enzymes, suggesting the existence of multiple pools of these lipids. (6) Addition of excess non-radiolabelled ATP to nuclei pre-labelled with [gamma-32P]ATP resulted in an initial increase in the label in PtdIns(4,5)P2, implying a precursor product relationship between the radiolabelled pools of PtdIns(4)P and PtdIns(4,5)P2. This was confirmed by analysis of the incorporation of 32P into the 4'-phosphate group of PtdIns(4)P and the individual 4'- and 5'-phosphate groups of PtdIns(4,5)P2. The data from these experiments also indicated that PtdIns(4,5)P2 can be produced from a pre-existing pool of PtdIns(4)P, as well as de novo from PtdIns. (7) Taken together our data suggest that isolated rat-liver nuclei have an intranuclear inositol lipid metabolism mechanism utilizing enzymes and substrates equivalent to those found in cytosol and plasma membrane, and that there may be some, but not complete, compartmentalization of the components of the nuclear inositol cycle. PMID- 9359433 TI - Expression of placental alkaline phosphatase does not correlate with IgG binding, internalization and transcytosis. AB - The human homologue of FcRn, an IgG Fc receptor expressed in rat villous syncytiotrophoblasts, might be involved in IgG transfer from the maternal to the fetal circulation. However, because the receptor does not bind IgG at the physiological pH of the maternal blood (pH 7.4), FcRn is probably not involved in the initial uptake of IgG. A role in IgG internalization has been suggested for placental alkaline phosphatase (PLAP), which is highly expressed on the apical surface of syncytiotrophoblasts. To determine whether PLAP does indeed have a role in IgG uptake, we analysed the ability of PLAP to bind, internalize and transcytose IgG in BeWo choriocarcinoma cells endogenously expressing the protein, or in Madin-Darby canine kidney (MDCK) cells transfected with the PLAP cDNA. Although PLAP expression in MDCK cells resulted in increased IgG binding to intact cells, binding was not correlated with the level of PLAP expressed in the different cell lines. Furthermore our findings do not support a role for PLAP in IgG endocytosis or transcytosis. PMID- 9359432 TI - Glypican-3 is a binding protein on the HepG2 cell surface for tissue factor pathway inhibitor. AB - Tissue factor pathway inhibitor (TFPI) is a primary regulator of the initiation of blood coagulation. TFPI is internalized and degraded by HepG2 cells through the low-density-lipoprotein receptor-related protein (LRP) but also binds another molecule present on the cell surface at approx. 10-fold the abundance of LRP [Warshawsky, Broze and Schwartz (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 6664 6668]. When HepG2 cells are washed with heparin or dextran sulphate, a substance that binds TFPI is removed from the cell surface and can be detected in a slot blot assay. Preincubation with trypsin destroys the reactivity of the TFPI binding component in the slot-blot assay, suggesting that it is a protein. In addition, when the sulphation of glycosaminoglycans (GAGs) is prevented by growing the HepG2 cells in the presence of 30 mM sodium chlorate, TFPI binding is unaffected, whereas the binding of bovine lipoprotein lipase, a protein known to associate with cell-surface GAGs, falls to 50% of control levels. Dextran sulphate washes of HepG2 cells grown in sodium chlorate have an equal reactivity in slot-blot experiments to that of non-treated cells, suggesting that GAGs are not totally responsible for the binding activity observed. By using the slot blot to follow binding activity and conventional protein purification techniques, a protein species that migrates at 40 kDa after reduction was identified in the HepG2 cell wash. The binding of this protein to TFPI was confirmed with immobilized TFPI. Amino acid sequence analysis identified this protein species as a proteolytic fragment of glypican-3 (also called OCI-5), a member of the glypican family of glycosylphosphatidylinositol-anchored proteoglycans. PMID- 9359434 TI - Characterization of chicken-liver glutathione S-transferase (GST) A1-1 and A2-2 isoenzymes and their site-directed mutants heterologously expressed in Escherichia coli: identification of Lys-15 and Ser-208 on cGSTA1-1 as residues interacting with ethacrynic acid. AB - Escherichia coli-expressed chicken-liver glutathione S-transferase, cGSTA1-1, displays high ethacrynic acid (EA)-conjugating activity. Molecular modelling of cGSTA1-1 with EA in the substrate binding site reveals that the side chain of Phe 111 protrudes into the substrate binding site and possibly interacts with EA. Replacement of Phe-111 with alanine resulted in an enzyme (F111A mutant) with a 4.5-fold increase in EA-conjugating activity (9.2 mmol/min per mg), and an incremental Gibbs free energy (DeltaDeltaG) of 4.0 kJ/mol lower than that of the wild-type cGSTA1-1. Two other amino acid residues that possibly interact with EA are Ser-208 and Lys-15. Substitution of Ser-208 with methionine generated a cGSTA1-1(F111AS208M) double mutant that has low EA-conjugating activity (2.0 mmol/min per mg) and an incremental Gibbs free energy of +3.9 kJ/mol greater than the cGSTA1-1(F111A) single mutant. The cGSTA1-1(F111A) mutant, with an additional Lys-15-to-leucine substitution, lost 90% of the EA-conjugating activity (0.55 mmol/min per mg). The Km values of the cGSTA1-1(F111A) and cGSTA1-1(F111AK15L) mutants for EA are nearly identical. The wild-type cGSTA2-2 isoenzyme has a low EA-conjugating activity (0.56 mmol/min per mg). The kcat of this reaction can be increased 2. 5-fold by substituting Arg-15 and Glu-104 with lysine and glycine respectively. The KmEA of the cGSTA2-2(R15KE104G) double mutant is nearly identical with that of the wild-type enzyme. Another double mutant, cGSTA2 2(E104GL208S), has a KmEA that is 3.3-fold lower and a kcat that is 1.8-fold higher than that of the wild-type enzyme. These results, taken together, illustrate the interactions of Lys-15 and Ser-208 on cGSTA1-1 with EA. PMID- 9359435 TI - Aging of di-isopropyl-phosphorylated human butyrylcholinesterase. AB - Organophosphate-inhibited cholinesterases can be reactivated by nucleophilic compounds. Sometimes phosphylated (phosphorylated or phosphonylated) cholinesterases become progressively refractory to reactivation; this can result from different reactions. The most frequent process, termed 'aging', involves the dealkylation of an alkoxy group on the phosphyl moiety through a carbocation mechanism. In attempting to determine the amino acid residues involved in the aging of butyrylcholinesterase (BuChE), the human BuChE gene was mutated at several positions corresponding to residues located at the rim of the active site gorge and in the vicinity of the active site. Mutant enzymes were expressed in Chinese hamster ovary cells. Wild-type BuChE and mutants were inhibited by di isopropylfluorophosphate at pH 8.0 and 25 degrees C. Di-isopropyl-phosphorylated enzymes were incubated with the nucleophilic oxime 2-pyridine aldoxime methiodide and their reactivatability was determined. Reactivatability was expressed by the first-order rate constant of aging and/or the half-life of aging (t12). The t12 was found to be of the order of 60 min for wild-type BuChE. Mutations on Glu-197 increased t12 60-fold. Mutation W82A increased t12 13-fold. Mutation D70G increased t12 8-fold. Mutations in the vicinity of the active site serine residue had either moderate or no effect on aging; t12 was doubled for F329C and F329A, increased only 4-fold for the double mutant A328G+F329S, and no change was observed for the A328G mutant, indicating that the isopropoxy chain to be dealkylated does not directly interact with Ala-328 and Phe-329. These results were interpreted by molecular modelling of di-isopropylphosphorylated wild-type and mutant enzymes. Molecular dynamics simulations indicated that the isopropyl chain that is lost interacted with Trp-82, suggesting that Trp-82 has a role in stabilizing the carbonium ion that is released in the dealkylation step. This study emphasized the important role of the Glu-197 carboxylate in stabilizing the developing carbocation, and the allosteric control of the dealkylation reaction by Asp-70. Indeed, although Asp-70 does not interact with the phosphoryl moiety, mutation D70G affects the rate of aging. This indirect control was interpreted in terms of change in the conformational state of Trp-82 owing to internal motions of the Omega loop (Cys-65-Cys-92) in the mutant enzyme. PMID- 9359436 TI - Molecular cloning and expression of a chloride channel-associated protein pICln in human young red blood cells: association with actin. AB - We report the cloning and sequencing from human reticulocytes of cDNA coding for the Cl- channel-associated protein, pICln. Human reticulocyte pICln (HRpICln) cDNA encodes a protein (predicted molecular mass 26293Da) identical with human non-pigmented ciliary epithelial cell pICln. By using full-length HRpICln cDNA (approx. 1.2 kb) to probe human lymphocyte metaphase-chromosome spreads, the location of the human ICln gene was mapped to 11q13 by fluorescence in situ hybridization analysis. Polyclonal antibodies to recombinant HRpICln detected bands at approx. 43 kDa and approx. 37 kDa in both normal (AA) and sickle (SS) red blood cell (RBC) ghost membranes. In SS ghosts, and in ghosts from a patient with autoimmune haemolytic anaemia with 9.8% reticulocytes, the amount of HRpICln was increased compared with AA ghosts, suggesting that the expression or membrane assembly of HRpICln is cell age-dependent. Laser scanning confocal fluorescent microscopy immunolocalized HRpICln largely to the RBC membrane. The increased staining intensity of HRpICln in a reticulocyte-enriched AA RBC density-separated fraction is consistent with a dependence of HRpICln membrane content on cell age. HRpICln and beta-actin form stable complexes in vivo, demonstrated with the yeast two-hybrid system. Low-ionic-strength extraction of ghost membranes, which results in the extraction of the spectrin-actin cytoskeleton, also results in the extraction of HRpICln, consistent with the possibility for the association of these proteins in RBCs in vivo. The results presented here establish the presence of the Cl- channel-associated protein, pICln, in human RBCs, and raises the possibility that this protein has a role in RBC Cl- transport and volume regulation in young RBCs. Moreover the association of RBC pICln with actin offers a model in which to test interactions between RBC ion channels and the cytoskeleton. PMID- 9359438 TI - Timing of insults causing abnormal outcome in preterm infants 1989-1992. AB - OBJECTIVE: The purpose of this study was to clarify the influence of timing of brain insults causing abnormal outcome in preterm infants. METHODS: One hundred and thirty-one preterm infants were examined. The timing of brain insult was estimated from EEG or clinical findings. Development was assessed until a corrected age of 48 months. RESULTS: 39% and 4% of infants, respectively, born before and after the 28-week time point subsequently died (P < 0.05). Abnormal development was observed in 16% of the first group and 13% of the second (N.S.). None of those born before 28 weeks showed intrauterine injuries while nine of the infants which were born after this time showed intrauterine injuries (P < 0.05). Fetal distress was noted in all infants suffering neonatal death born after 28 weeks. CONCLUSION: Intrauterine brain insult was concluded to be the cause of neonatal death or abnormal development in many infants born after 28 weeks. PMID- 9359437 TI - Cloning and characterization of a complementary DNA for a thyroid hormone responsive protein in mature rat cerebral tissue. AB - A gene responsive to thyroid hormone (TH) has been identified in the adult rat brain cerebral tissue. A cDNA probe differentially expressed in euthyroid, hypothyroid and hyperthyroid rat cerebral tissue, generated by reverse transcriptase-PCR differential display of mRNA, was used to screen the rat brain cDNA library. A 3.4 kb positive clone hybridized in Northern blots with a 3.8 kb mRNA that proved to be TH responsive (THR). The remaining coding sequence and a part of the 5' untranslated region of this cDNA were obtained by 5' rapid amplification of cDNA ends. The deduced amino acid sequence revealed that THR protein (THRP), a 68 kDa moiety, has 83% sequence similarity with c-Abl interactor protein (Abi-2), which is a substrate for tyrosine kinase activity of c-Abl. The extensive similarity between the two proteins suggests a potential role for THRP as a substrate for c-Abl. Northern analysis showed that the expression of THR mRNA in hyperthyroid rats is 6-fold that in euthyroid rats. There is also a 4-6-fold increase in the concentration of THRP, as analysed by Western analysis. Owing to the extensive similarity between rat THRP and human Abi-2, a polyclonal anti- (human Abi-2) antibody was successfully used for Western analysis of proteins from the rat tissues. The observed increase in both the mRNA and the protein did not decline after beta-adrenergic system blockade with propranolol, suggesting that the action of TH on the expression of this gene is not mediated through the beta-adrenergic system. Immunohistochemical studies revealed that neuronal cells were particularly rich in THRP. Both THR mRNA and THRP are rapidly induced in vivo after intravenous administration of thyroxine. Tissue distribution studies indicated that the cerebral tissue was particularly enriched with THR mRNA and 68 kDa THRP. A cDNA clone for a THR gene could provide a useful tool to study the molecular mechanisms of TH effects on cerebral tissue in adult animals. PMID- 9359440 TI - External cephalic version at term with tocolysis and vibroacoustic stimulation. AB - OBJECTIVE: This study compared the obstetric outcome in women who had external cephalic version (ECV) for breech presentation after 36 weeks gestation with those who did not, to see whether ECV reduces breech deliveries and cesarean section rates with reduced complications. METHOD: External cephalic version was attempted in 200 women (study group) with the use of tocolysis and vibroacoustic stimulation. The control group (ECV not attempted) comprised of 278 women with breech presentation after 36 weeks. RESULT: The cesarean section rate was 14.0% in the successful version group compared with 55.2% in the unsuccessful version group. The overall cesarean section rate in the study group was 32.5%. In the control group of 278, the fetus remained in the breech presentation in labor in 269 women with a cesarean section rate of 51.4% which was not different from the unsuccessful version group (55.2%). CONCLUSION: This study supports the randomized trials conducted earlier in that ECV after 36 weeks gestation reduced the number of breech deliveries and cesarean sections (32.5% in the study group compared with 51.4% in the control group) (P > 0.00001). PMID- 9359439 TI - Cervicovaginal washing prolactin assay in prediction of preterm delivery. AB - OBJECTIVE: Our purpose was to determine the utility of cervicovaginal washing prolactin assay in prediction of preterm birth in women without rupture of membranes. METHODS: Sixty-six women with normal singleton pregnancy were submitted to cervicovaginal washing and serum prolactin assays. The latency period to delivery and gestational age at admission and at delivery were also recorded. According to uterine contractions and obstetrical history regarding the previous preterm delivery, the pregnant women were divided into 4 groups: 18 symptomatic (group 1) and 15 asymptomatic (group 2) pregnancies who had previously had preterm delivery, and 18 symptomatic (group 3) and 15 asymptomatic (group 4) pregnancies without a history of prior preterm delivery were enrolled in the study. RESULTS: The cervicovaginal washing prolactin concentrations were significantly higher in groups 1 and 3 than in group 4 (P < 0.0083). With respect to the latency period to delivery and the birth weeks, groups 2 and 4 were significantly higher than groups 1 and 3 (F < 0.0001). In the evaluation of the whole group, a significant negative correlation was observed both between cervicovaginal washing prolactin concentrations and the lapsed times to delivery, and the gestational ages at delivery. The finding of a cervicovaginal washing prolactin value exceeding 50 ng/ml in the 12 days preceding preterm delivery had sensitivity, specificity, positive and negative predictive values of 65%, 95%, 86%, and 81%, respectively. CONCLUSIONS: A cervicovaginal washing prolactin value more than 50 ng/ml precedes preterm delivery within 12 days at > 29 weeks. The easy application, the good feasibility, the success in identifying pregnancies at risk for preterm labor, and the cost effectiveness suggests cervicovaginal washing prolactin assay as a biochemical marker for preterm delivery. PMID- 9359441 TI - Pregnancy outcome beyond 41 weeks gestation. AB - OBJECTIVE: To determine maternal and perinatal morbidity and the spontaneous labor rate beyond 41 weeks of gestation. METHOD: Patients with uncomplicated pregnancy were recruited at 41 weeks and screened for fetal or maternal well being. Following observation between 41 and 42 weeks, patients were randomized to either serial monitoring by cardiotocography and measurement of amniotic fluid index, or to immediate induction. Comparisons were made using the chi(2) test. Results after 42 weeks were analyzed according to intention at randomization. RESULTS: Morbidity was not increased before 42 weeks. After 42 weeks, the cesarean section rate and incidence of meconium below the vocal cords were increased in monitored patients. The median gestational age in patients who were monitored was 298.5 (294-321) days. In patients observed from 41 weeks, 91.6% labored spontaneously. CONCLUSION: It is reasonable to observe uncomplicated pregnancy until 42 weeks with adequate monitoring. After 42 weeks, induction of labor is preferred. PMID- 9359442 TI - Routine colposcopic evaluation of patients with persistent inflammatory cellular changes on Pap smear. AB - OBJECTIVE: To assess the rate of undetected dysplasia in patients with two consecutive reports of inflammatory cellular changes without atypia on Pap smears despite anti-inflammatory therapy. METHOD: A prospective randomized study. RESULT: 2798 premenopausal non-pregnant patients were evaluated by Pap smears of the cervix. Of these, 397 (14.2%) were reported as 'inflammatory cellular changes'. A total of 238 (8.5%) had persistent inflammatory changes without atypia despite the anti-inflammatory therapy. Fourteen patients refused colposcope. The mean age and parity of the remaining 224 patients were 30.2 +/- 6.3 (18-46) and 1.7 +/- 2.3 (0-6), respectively. When these patients underwent colposcopically-directed biopsies of the cervix, in 51 (22.7%) patients human papillomavirus lesions, dysplasia and in situ cancer were noted. Mean age, parity, age of marriage, prevalence of smoking and contraceptive methods of the two groups of patients (173 vs. 51) did not show statistically significant differences. CONCLUSION: Colposcopically-directed biopsies of the cervix are indicated even when inflammatory cellular changes without atypia persist despite therapy. PMID- 9359443 TI - Sclerotherapy--an adjuvant therapy to endometriosis. AB - OBJECTIVE: Recurrent endometriomas after medical or surgical treatment is a difficult clinical problem for those patients who wish to perform ovulation induction. Therefore we tried to investigate the efficacy of sclerotherapy as an adjuvant management before ovulation induction to preserve more ovarian tissue for folliculogenesis in ART program. METHODS: Thirty-two patients with persistent or recurrent endometrioma after surgical or medical treatment were included in this study. Transvaginal ultrasound needle guided aspiration of the cyst followed by tetracycline instillation was performed before ovulation induction. RESULTS: There is an encouraging clinical pregnancy rate of 34.37%. Also, there is a disappointing recurrent rate of 46.87% in 12 months follow-up course. CONCLUSION: The increased interest in cost-effective outpatient therapy and the expected difficulty in surgical treatment of recurrent endometriomas made aspiration and sclerotherapy of endometrioma an attractive option before ovulation induction. PMID- 9359445 TI - Torsion of the pregnant uterus at term. PMID- 9359444 TI - Reliability of preoperative evaluation of prognostic factors in endometrial carcinoma. AB - OBJECTIVE: To define the accuracy of preoperative evaluation of prognostic factors in detecting patients with low risk of node metastasis in which different surgical approaches can be proposed. SUBJECTS: Seventy-five patients with a histologically proven endometrial carcinoma were considered in this study. METHODS: All the patients underwent a preoperative evaluation of grading (G), and myometrial invasion (M) by endometrial biopsy and transvaginal ultrasound (TVS). In 41 patients preoperative ploidy of carcinoma cells (P) was determined by flow cytometry. Pre-surgical G, M and P were then compared with surgical specimens. We considered 'low risk', patients with no or moderate myometrial invasion, well differentiated histological grading and diploid DNA. RESULTS: We were able to identify 19/23 (82.6%) low risk cases. Correct identification of high risk patients was obtained in 49/52 (94%) patients. In three low risk patients, correctly diagnosed preoperatively, the final FIGO stage was IIIA (two for adnexal involvement and one for positive peritoneal washing). CONCLUSIONS: Our findings suggest that it is possible to detect preoperatively patients with a low risk of node metastasis. Alternative surgical approaches, i.e. vaginal surgery, can be taken into account in such patients. PMID- 9359446 TI - Conservative approach in unruptured cornual pregnancy with a live fetus. PMID- 9359447 TI - Medical management of placenta accreta. PMID- 9359448 TI - Expectant management and methotrexate treatment of persistent ectopic pregnancy following laparoscopic salpingectomy. PMID- 9359449 TI - Anemia in pregnancy: case control study of risk factors. PMID- 9359450 TI - Fibroma of the vulva and uterine leiomyoma. PMID- 9359452 TI - How accurate is the hysteroscopic diagnosis of endometrial polyp? PMID- 9359451 TI - Extrapelvic endometriosis in Nigeria. PMID- 9359453 TI - Successful hormonal treatment of pulmonary parenchymal endometriosis. PMID- 9359454 TI - Munchausen's syndrome: gynecologic trauma with Stanley-knife blades. PMID- 9359455 TI - ACOG educational bulletin. Lower urinary tract operative injuries. Number 238, July 1997. American College of Obstetricians and Gynecologists. PMID- 9359456 TI - ACOG practice patterns. External cephalic version. Number 4, July 1997. American College of Obstetricians and Gynecologists. PMID- 9359457 TI - Diagnostic laparoscopy for acute pelvic pain. Number 25, July 1997. Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9359458 TI - Isoquinoline neurotoxins in the brain and Parkinson's disease. AB - Parkinson's disease is thought to be caused by some unknown endogenous or exogenous factors interacting with genetic dispositions. 1-Methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) is an exogenous neurotoxin producing parkinsonism in humans, monkeys and various animals as the result of monoamine oxidase type B (MAO-B)-catalyzed conversion of it to the 1-methyl-4-phenyl pyridinium ion (MPP+), which selectively kills the nigrostriatal dopaminergic neurons. Various isoquinoline derivatives were found in the brain of patients with Parkinson's disease. Isoquinoline derivatives have neurochemical properties similar to those of MPTP and they are considered to be the endogenous neurotoxins which cause Parkinson's disease. Among them, tetrahydroisoquinoline (TIQ), 1 benzyl-TIQ, and (R)-1,2-dimethyl-5,6-dihydroxy-TIQ [(R)-N-methyl-salsolinol)] have the most potent neurotoxicity. TIQs, like MPTP, may be activated via N methylation by N-methyltransferase and oxidation by MAO. TIQs as well as MPP+ inhibit complex I of the electron transport system in mitochondria, thereby reducing ATP formation and producing oxygen radicals. Although the properties of TIQs are similar to those of MPTP, the neurotoxicity of TIQs is weaker than that of MPTP. Since Parkinson's disease is a slowly progressing neurodegenerative disease, long term neurotoxic effects of IQs remain to be further examined in primates. PMID- 9359459 TI - Suppression of pair-pulse depression of the population spike in the dentate gyrus during carbachol-induced theta-like activity in guinea pig hippocampal slices. AB - In pair-pulse stimulation experiments, pair-pulse depression (PPD) of the population spike (PS) occurred at intervals shorter than 20 ms in the dentate gyrus in guinea pig hippocampal slices. Application of 50 microM carbachol resulted in an increase in the test PS amplitude and caused suppression of PPD. This suppression was antagonized by atropine sulfate, a muscarinic receptor antagonist. Carbachol at 50 microM induced intermittent bursts of theta-like activity (TLA). We compared the pair-pulse index (PPI) during TLA with that in a rest period between bursts of TLA. The PPI was defined as the ratio of the amplitude of the test PS to that of the conditioning PS. The PPI during TLA were significantly larger than that during the rest period, although there were no significant differences in the conditioning PS amplitude and the test pEPSP slope. When TLA was induced, the PPI during the rest period was increased by bicuculline. The PPI during TLA did not change significantly with the drug. The increase by bicuculline in the PPI during the rest period was caused by increase in the test PS amplitude. PPD can occur due to inhibition of granule cell activity by inhibitory neurons. Our findings suggest that the action of inhibitory neurons on granule cell activity is suppressed by activation of muscarinic receptors, with stronger suppression during TLA than during the rest period between bursts of TLA. PMID- 9359460 TI - Etorphine inhibits cell growth and induces apoptosis in SK-N-SH cells: involvement of pertussis toxin-sensitive G proteins. AB - Opiates have been used extensively in the treatment of pain but with the severe side effect of addiction, which is believed to be related to opiates' direct (primary) or indirect (secondary) neurotoxicity. In this study, the effects of opioids on cell growth and apoptosis have been examined in human neuroblastoma cell line SK-N-SH. Etorphine, a wide-spectrum and potent agonist of opioid receptors, was found to significantly inhibit cell growth and to induce apoptosis. The inhibitory and apoptotic activities of etorphine followed a dose- and time-dependent manner. The more specific agonists of opioid receptors such as morphine, [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAGO), [D-Pen2, D-Pen5] enkephalin (DPDPE), dynorphin A and nociceptin/orphanin FQ did not show similar toxic activities under the same conditions. In addition, the effects of etorphine could not be blocked by the opioid receptor antagonist naloxone, suggesting that the effects of etorphine might not be mediated by a classical opioid receptor. However, pretreatment of SK-N-SH cells with pertussis toxin (PTX) blocked the inhibition of cell growth and apoptosis induced by etorphine, indicating the involvement of PTX-sensitive G proteins in the processes. It was also shown that etorphine-induced apoptosis was prevented by actinomycin D (AD) and interleukin 1beta converting enzyme inhibitor I. Interestingly, etorphine was similarly potent to inhibit growth of pheochromocytoma (PC12) cells but less effective in SH-SY5Y neuroblastoma cells and C6 glioma cells. We propose that inhibition of cell growth and induction of apoptosis may be one mechanism of opioid neurotoxicity. PMID- 9359461 TI - Congo red reverses amyloid beta protein-induced cellular stress in astrocytes. AB - An amyloid-binding dye Congo red has been reported to prevent the neurotoxic effect of Alzheimer's amyloid beta protein (Abeta). In the present study, we investigated the effect of Congo red in cultured rat cortical astrocytes. Abeta (1 nM-10 microM) did not cause cell death, but potently inhibited the cellular redox activity as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction assay. Congo red (0.2-20 microM), when added together with or prior to Abeta, significantly blocked Abeta-induced inhibition of redox activity. Furthermore, when Congo red was added after treatment with Abeta, the inhibited redox activity was restored to normal, indicating that Congo red can reverse Abeta-induced cellular stress. The reversing effect of Congo red cannot be explained by the inhibition of Abeta fibril formation and suggests a novel aspect of the interaction of Congo red with Abeta. PMID- 9359462 TI - Single neurons in the spinal trigeminal and dorsal column nuclei project to both the cochlear nucleus and the inferior colliculus by way of axon collaterals: a fluorescent retrograde double-labeling study in the rat. AB - A number of single neurons in the spinal trigeminal nucleus (STN) and the dorsal column nucleus (DCN) were found to project simultaneously to the cochlear nucleus (CoN) and the external cortex of the inferior colliculus (ICe) by way of axon collaterals. Each rat was injected with Fluoro-Gold (FG) into CoN on one side and with tetramethylrhodamine-dextran amine (TMR-DA) into ICe on the side ipsilateral or contralateral to the FG injection. In these rats, a number of neuronal cell bodies in DCN and the interpolar and caudal subnuclei of STN were double-labeled retrogradely with both FG and TMR-DA, mainly on the side ipsilateral to the FG injection into CoN. These neurons in STN and DCN might mediate somatosensory inputs simultaneously to the two lower brainstem nuclei, CoN and ICe, which constitute the relays of the auditory pathway. PMID- 9359463 TI - Intradermal 5-HT induces Fos expression in rat dorsal horn neurons not via 5-HT3 but via 5-HT2A receptors. AB - We investigated the effects of peripherally administered 5-HT on the secondary neurons in the spinal cord of rats using Fos-like immunoreactivity (FLI) as a marker of neuronal activation. The intradermal administration of 5-HT (30, 60 microg) induced a large number of FLI neurons in the ipsilateral dorsal horn. In animals given 5-HT2A receptor agonists (DOI: 0.28 to 2.8 micromol/kg, alpha methyl 5-HT: 0.28 to 2.8 micromol/kg) intradermally, immunoreactive neurons were evoked in the same manner as those given 5-HT. Other agonists, including 5-HT3 receptor agonists (m-CPG: 16 to 32 micromol/kg, 2-methyl 5-HT: 0.0028 to 2.8 micromol/kg), did not induce FLI neurons at any dose examined. Furthermore, 5 HT2A receptor antagonist (ketanserin: 1 mg/kg, i.p.) suppressed the expression of FLI in the dorsal horn caused by peripheral 5-HT, but 5-HT3 receptor antagonist (tropisetron: 1 mg/kg, i.p.) did not. These findings suggest that the 5-HT induced nociceptive response is mediated by 5-HT2A receptors in the periphery. PMID- 9359464 TI - GABA(B) receptor activation of Purkinje cells in cerebellar slices. AB - The metabotropic GABA(B) receptors are densely represented in the molecular layer of the cerebellar cortex which contains the dendritic tree of the Purkinje cells (PCs). We report here the results obtained by applying Baclofen, the specific GABA(B) agonist, to PCs recorded intrasomatically in cerebellar slices. Diluted in the perfusion solution or applied by pressure to the molecular layer near to the recorded cell, Baclofen dose-dependently inhibited the PCs as seen by the suppression of Na and Ca dependent action potentials accompanied by a variable membrane hyperpolarization. The weak hyperpolarization was interpreted as due to the dendritic localization of the receptors. These results concerned postsynaptic receptor sites since they persisted after bath applied TTX blocking presynaptic activity. They also persisted in the presence of bicuculline, the GABA(A) antagonist, but they were reduced by bath application of 2-OH saclofen and CGP55845A, both being GABA(B) receptor antagonists. Current clamp experiments revealed a conductance increase with an equilibrium potential consistent with a K+ channel opening. The conclusions were reached that GABA inhibition of the PCs is mediated by GABA(B) receptors in the dendrites and GABA(A) receptors in the soma and dendrites. Therefore, the GABA released by stellate cells modulate PC activity through two inhibitory mechanisms. PMID- 9359465 TI - A mathematical model that reproduces vertical ocular following responses from visual stimuli by reproducing the simple spike firing frequency of Purkinje cells in the cerebellum. AB - A mathematical model that accurately reproduces eye movements from visual stimuli and incorporates intermediate neural signals is useful for quantitative analysis of the neural mechanisms involved in transforming visual stimuli to eye movements. Here we describe a mathematical model consisting of two systems: a non linear system that relates retinal slip to simple spike firing frequency of Purkinje cells in the ventral paraflocculus (VPFL) and a linear system that relates VPFL simple spike firing frequency to eye movement. This model accurately reproduced the firing frequency of Purkinje cells and ocular following responses from visual stimulation paradigms used in physiological experiments. PMID- 9359467 TI - Differential expression of mRNA for leptin receptor isoforms in the rat brain. AB - Leptin plays an important role in the control of food intake and energy metabolism by interacting with its receptor (OB-R) in the brain. Several alternatively spliced isoforms of OB-R have been identified. To study the expression patterns and the potential biological function of these OB-Rs in the brain, the distribution of mRNA encoding OB-R isoforms was examined by in situ hybridization. In agreement with previous studies, strong signals for OB-R mRNA were detected in the hypothalamus, thalamus and choroid plexus. In addition, intense signals were observed in several other brain areas including piriform cortex, granule cell layer of the cerebellum and substantia nigra. With isoform specific probes, a differential expression pattern of OB-Rs was revealed: OB-Ra and OB-Rb, but not OB-Rc and OB-Rf, are abundantly expressed in the hypothalamus, whereas OB-Ra, OB-Rc and OB-Rf, but not OB-Rb, are significantly expressed in the choroid plexus. The preferential expression of OB-Rb in the hypothalamus is in support of its role in mediating the satiety effect of leptin. The co-expression of OB-Ra with OB-Rb in the hypothalamus may suggest a possible interaction between the two isoforms. Finally, the detection of OB-R mRNA in a number of other brain regions may indicate the involvement of leptin in additional as yet undefined physiological functions. PMID- 9359466 TI - Differential effects of phorbol ester on AMPA and NMDA components of excitatory postsynaptic currents in dentate neurons of rat hippocampal slices. AB - Protein kinase C (PKC) is present abundantly in the mammalian central nervous system, and is involved in a variety of neuronal functions. Phorbol esters mimic the role of diacylglycerol, the physiological activator of PKC. We examined effects of phorbol 12,13-diacetate (PDAc) on excitatory synaptic transmission in neurons in the dentate granule cell layer of rat hippocampal slices using the whole-cell patch clamp technique. Excitatory postsynaptic currents (EPSCs) evoked by stimulation of the perforant path (pp) consisted of AMPA and NMDA receptor mediated components. The application of PDAc potentiated both components of the EPSC, but the effect was more pronounced on the NMDA component. The potentiating effect of PDAc on the NMDA component was dependent on the membrane potential, being most prominent at - 31 and -51 mV. Omega-agatoxin-IVA, a P-type Ca2+ channel blocker, suppressed both AMPA and NMDA components to a similar extent by reducing transmitter release. However, when the PDAc-potentiated AMPA component was reduced to the control level by applying omega-agatoxin-IVA, a substantial potentiation on the NMDA component remained. These results suggest that the potentiation of the NMDA component of the EPSC by PDAc is caused partly by a postsynaptic mechanism in the dentate neurons. PMID- 9359468 TI - Transforming growth factor-alpha stimulates insulin-like growth factor binding protein-4 (IGFBP-4) expression and blocks follicle-stimulating hormone regulation of IGFBP-4 production in rat granulosa cells. AB - The ability of TGF-alpha to regulate insulin-like growth factor binding protein-4 (IGFBP-4), was investigated. Primary cultures of rat granulosa cells (GC) were grown in serum-free medium with rat (r) TGF-alpha and/or rFSH, and secreted IGFBP 4 protein and its steady state mRNA levels were measured by Western immunoblotting and Northern blotting, respectively. Control (untreated) cells secreted IGFBP-4 spontaneously, and the levels were increased by rTGF-alpha in a dose- and time-dependent manner. rTGF-alpha abolished FSH-induced IGFBP-4 protease activity and suppressed FSH-dependent effects on IGFBP-4 production. IGFBP-4 mRNA levels were decreased and increased by FSH and TGF-alpha, respectively, and TGF-alpha blocked the FSH effects. These results demonstrate that TGF-alpha is a potent stimulator of IGFBP-4 expression in rat GC and can overcome the regulatory effects of FSH on IGFBP-4 production. The consequence of these TGF-alpha effects is a marked, sustained increase in the levels of IGFBP-4 in the microenvironment. PMID- 9359469 TI - Identification of neuropeptides in the midgut of parasitized insects: FLRFamides as candidate paracrines. AB - Parasitism of Manduca sexta (Lepidoptera: Sphingidae) larvae by the braconid wasp Cotesia congregata (Hymenoptera: Braconidae) leads to accumulation of peptides in host neurons and neurosecretory cells of the central nervous system (CNS) and neurons and endocrine/paracrine cells of the midgut. This accumulation has now facilitated the characterization of two new members of the FLRFamide family from midguts of parasitized larvae. The peptides, given the names F24 and F39, are 24 and 39 amino acids in length with the sequences VRDYPQLLDSGMKRQDVVHSFLRFamide and YAEAAGEQVPEYQALVRDYPQLLDSGMKRQDVVHSFLRFamide. The sequence of F24 is identical to the C-terminal 24 amino acids of F39. The C-terminal 10-mer of each is identical to a previously characterized decapeptide neurohormone (F10). This sequence is preceded by a potential processing site. In nonparasitized insects F39 was present at several-fold the amount of F24. In parasitized insects F24 and F39 accumulate in the middle and posterior regions of the midgut, which are enriched in endocrine/paracrine cells reacting with FLRFamide antisera. In the combined brain and subesophageal ganglion F39 was not detected and the amount of F24 never exceeded 2 fmol per Br/SEG. Of the three peptides, only F10 was found in the hemolymph. Thus, F24 and F39 may be intermediates in the biosynthesis of F10 and may themselves be released locally from endocrine/paracrine cells in the midgut epithelium. PMID- 9359470 TI - Impairment of ATP-induced Ca2+ -signalling in human thyroid cancer cells. AB - Extracellular nucleotides like ATP that activate the Ca2+ -phosphatidylinositol (PI) signalling pathway have been suggested to participate in the regulation of normal human thyroid function. We examined, whether P2y-purinergic receptors are expressed on human thyroid cancer cells and whether post-receptor Ca2+ signalling is altered by malignant transformation. Extracellular ATP caused a biphasic increase in cytosolic free Ca2+ ([Ca2+]i) in normal human thyrocytes and in human follicular (FTC) and papillary (PTC) thyroid carcinoma cells. In FTC and PTC cell lines the dose-response curves for ATP-induced changes in [Ca2+]i were shifted to the right when compared with normal thyrocytes, whereas in undifferentiated thyroid carcinoma (UTC) cells even high concentrations of ATP (500 microM) failed to stimulate a rise in [Ca2+]i. By contrast, ATP stimulated inositol 1,4,5 trisphosphate (IP3) formation and capacitative Ca2+ entry was operational as judged by thapsigargin in normal thyrocytes and all thyroid cancer cells. Thus, P2y-purinergic receptors are expressed on thyroid tumor cells independent of degree of differentiation. In UTC cells, however, impairment in the Ca2+ phosphatidylinositol (PI) signalling cascade occurs distal to the formation of IP3 and proximal to the activation of capacitative Ca2+ entry. Disturbed ATP induced Ca2+ -signalling and alterations in the Ca2+ -PI signalling cascade may contribute to decreased expression or loss of specific thyroid functions in thyroid cancer cells. PMID- 9359471 TI - Interleukin-1 beta inhibits progesterone accumulation in rat corpora luteal cell cultures in a mechanism dissociated from its effects on nitric oxide and prostaglandin E accumulation. AB - The objective of this study was to examine the effect of interleukin-1 beta (IL 1) on progesterone (P) biosynthesis and the potential intermediary involvement of prostaglandin (PG) E and nitric oxide (NO) in P accumulation in PMSG/hCG-primed rat corpora luteal (CL) cell cultures. Exposure of primed CL cells to IL-1 (10 ng/ml) for 48 h resulted in a 65-86% reduction (P < 0.01) in P accumulation concurrent with a 2-3.4-fold increase in PGE content, a 70% increase in PGF2 alpha content and a 1.9-3.3-fold increase in nitrite generation. These effects were abolished by the IL-1 receptor antagonist, suggesting specific IL-1 receptor mediated effects. Indomethacin, a cyclooxygenase inhibitor, abolished PGE and PGF2 alpha production and attenuated the basal (but not IL-1-stimulated) accumulation of P. N(G)-Nitro-L-arginine (NNLA), a competitive inhibitor of nitrite synthesis, slightly reduced basal P accumulation but had no effect on IL 1-induced suppression of P accumulation. NNLA reduced basal PGE accumulation and IL-1-stimulated PGE accumulation (55 and 61%, respectively). Transforming growth factor beta 1 (TGF-beta 1; 10 ng/ml) significantly attenuated the IL-1-stimulated PGE and NO production (61 and 42%, respectively), but did not affect the ability of IL-1 to suppress P accumulation. Thus, NO, PGF2 alpha and PGE are not obligatory intermediaries of IL-1-mediated suppression of P accumulation in rat CL, but are involved in basal P biosynthesis and NO seems to have a regulatory role in the biosynthesis of PGE. The present observations suggest a pleiotropic response of PMSG/hCG-primed CL cells to IL-1, characterized by an independent suppression of P accumulation and a concomitant increase in NO, PGF2 alpha and PGE generation. Since IL-1 attenuates P accumulation, these findings may imply a direct autocrine/paracrine function for IL-1 in the maintenance or the demise of rat CL. PMID- 9359472 TI - 17 beta-Estradiol-mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor: cell cycle effects. AB - Estrogen receptor (ER)-negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon (Ah)-responsiveness. Treatment of the stably transfected cells with 10 nM 17 beta-estradiol (E2) resulted in a significant inhibition (> 60%) of cell proliferation and DNA synthesis, which was blocked by 10(-7) M ICI 182 780. Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/G1 (from 68.8 to 89.4) and decreased cells in S (from 18.4 to 3.4) and G2/M (from 12.8 to 7.2) phases of the cell cycle. The effects of E2 on the major cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors, retinoblastoma protein (RB), E2F-1, and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells. The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis, including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h) and decreased E2F1 and PCNA protein levels. These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/G1 and inhibition of DNA synthesis. PMID- 9359474 TI - Dendritic cells as mediators of tumor-induced tolerance in metastatic melanoma. AB - Escape from immune surveillance is critical for tumor progression in metastatic melanoma. We assessed the function of melanoma-derived dendritic cells (DCs) in patients presenting simultaneously with responding (rM) or progressing (pM) melanoma metastases. These rare coincidences allowed us to compare syngeneically the function of tumor DCs. CD83+ DCs were purified freshly from large responding (rDCs) or progressing (pDCs) metastases following chemoimmunotherapy. rDCs were 5 times more potent inducers of allogeneic T-cell proliferation than the pDCs that were used as control. Phenotypic analysis showed a marked depression of CD86 expression on pDCs. Culture supernatants from pM showed production of Th2-type cytokines [interleukin-10 (IL-10)], whereas a Th1 pattern [IL-2, interferon-gamma (IFN-gamma), IL-12) predominated in rM. The IL-10 detected in progressing metastases was directly derived from melanoma cells. Culture supernatants from metastases applied to DC-supported allo-MLR assays suppressed T-cell responses by 50-75% in the case of pM, but not rM. Finally, in a co-stimulation-dependent anti CD3 tolerance assay, pDCs (but not rDCs) induced anergy in syngeneic CD4+ T cells. Anergy could be overcome by addition of IL-12 or IL-2. Our results show that melanoma-derived factors convert DC-antigen presenting cell function to tolerance induction against tumor tissue, changing tumor DCs to "silencers" of anti-tumoral immune responses. PMID- 9359473 TI - Regulation of cytochrome P450 cholesterol side-chain cleavage, 3 beta hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase type 1 and estradiol-17 beta-hydroxysteroid dehydrogenase mRNA levels by calcium in human choriocarcinoma JEG-3 cells. AB - In human placenta the cytochrome P450 side-chain cleavage (P450scc) and 3 beta hydroxysteroid dehydrogenase type 1 (3 beta-HSD-1) convert cholesterol and pregnenolone producing progesterone, whereas 17 beta-hydroxysteroid dehydrogenase type 1 (17 beta-HSD-1) mediates the interconversion of estrone and estradiol. We have examined the effects of calcium on phorbol ester- and cAMP-induced P450scc, 3 beta-HSD-1 and 17 beta-HSD-1 mRNAs in human JEG-3 cells. A23187 increased in a dose-dependent fashion in the 1.3 kb 17 beta-HSD-1 mRNA whereas a weaker increase followed by a gradual depletion effect of A23187 was observed on 3 beta-HSD-1 mRNA. No significant effect of A23187 on P450scc mRNA was observed. Using 0.50 microM of A23187 the induction of 3 beta-HSD-1 and 17 beta-HSD-1 mRNAs was maximum within about 6 h whereas P450scc mRNA levels stayed unaffected throughout the time-course period. The action of A23187 was synergistic on cAMP-stimulated 17 beta-HSD-1 mRNA levels, while in a dose-dependent manner A23187 progressively depleted 3 beta-HSD-1 and P450scc mRNA abundance probably by activation of a calcium-/calmodulin-dependent phosphodiesterase. On the phorbol 12-myristate, 13 acetate (PMA)-stimulated 3 beta-HSD-1, 17beta-HSD-1 and P450scc mRNA levels only the lowest concentration of A23187 potentialized the PMA effect on the 17 beta HSD-1 mRNA levels. Using thapsigargin (TG), a cell-permeable sesquiterpene lactone that releases calcium by inhibiting sarco/endoplasmic reticular calcium ATPase, our data indicated the presence in JEG-3 cells of TG-sensitive and TG insensitive calcium-ATPases regulating 3 beta-HSD-1 and 17 beta-HSD-1 mRNA levels. These results emphasized the complexity of calcium contribution with the protein kinase A and C pathways in the regulation of P450scc, 3 beta-HSD-1 and 17 beta-HSD-1 mRNA levels. In addition, the different sensitivity of these genes to calcium suggest they could be activated by different subclasses of PKCs. PMID- 9359475 TI - Social environment and prognosis of colorectal cancer patients: a French population-based study. AB - Colorectal cancer is a major public health problem in industrialised countries. Several studies have shown that social environment influences survival in cancer patients in many countries, but the causes remain unknown. In France, very little work has been done in this area. Our aim was to assess whether social environment influences survival of colorectal cancer patients in a well-defined French population and, if so, to what extent this could be explained by differences in stage at diagnosis or in treatment. The study population consisted of 1,642 colorectal cancer patients diagnosed between 1978 and 1987 in the French department of Calvados. Socio-demographic characteristics were assessed in terms of socio-professional category, place of residence (urban vs. rural) and distance from the place of residence to a specialised health-care centre. The relation between social environment, clinical factors and survival was studied using 2 multivariate methods (Cox model and relative survival method). Patients with poorer prognosis were found to be farmers of both sexes and individuals without occupation among males. Differences in survival were not explained entirely even when variations in stage at diagnosis and in treatment were taken into account. PMID- 9359476 TI - Anthropometric indices in relation to mammographic patterns among peri-menopausal women. AB - The relationship between body height, weight and body mass index and mammographic patterns was examined among 3,208 Norwegian women, aged 40-56 years, participating in the Third Tromso study. Standardized measurements of height and weight were recorded. Epidemiologic data were obtained through questionnaires. Mammograms were categorized into 5 groups based on anatomic-mammographic correlations. For analysis, patterns I-III were combined into a low-risk group and patterns IV and V into a high-risk group. Odd ratios (ORs), adjusted for menopausal status, age, parity, age at first birth, age at menarche and anthropometric measures, with 95% confidence intervals (CIs), were calculated. Body height was associated positively with high-risk patterns, while weight and body mass index were associated inversely with high-risk patterns. Women in the highest tertile of height were twice as likely (OR = 2.0, 95% CI 1.6-2.6) to have high-risk patterns compared with those in the lowest tertile, and women in the highest tertile of weight were 70% less likely (OR = 0.3, 95% CI 0.2-0.4) to have high-risk patterns compared with those in the lowest tertile. Associations with body mass index were similar to those with weight. All associations were present when stratified by menopausal status. Among post-menopausal women, the inverse associations between body weight and body mass index and high-risk patterns decreased with increasing number of years since menopause. Our results indicate that body height and weight are independently associated with the mammographic pattern among peri-menopausal women. We suggest that body height and weight are related to mammographic patterns through different mechanisms. PMID- 9359477 TI - Increased hyaluronidase levels in breast tumor metastases. AB - Hyaluronidase, a matrix-degrading enzyme, was assayed in extracts from breast primary tumors and regional metastases using a pool of human sera as a standard. Optimal activities of tumor extracts and serum were found for concentrations of 0.15-0.20 M NaCl in pH 3.8-4.0 buffer. In evaluating contamination by serum due to vascular proliferation, we expressed our results as the ratio of the entire activity (mU/l extract) on serum albumin content of tumors (g/l). Median (interquartile range) activities were 9.02 (6.04-14.34) for primary tumors and 37.36 (24.06-99.63) mU/g albumin for metastases. The difference was significant. Zymographic analysis showed that 3 bands of activity were detected which corresponded to 68, 53 and 49 kDa for tumoral hyaluronidase. The same pattern was observed for cellular extracts of breast cancer cell line CAL51, demonstrating that hyaluronidase detected in tumor extracts had mainly a cellular origin. Our results suggest that hyaluronidase may be involved in the metastatic process. PMID- 9359479 TI - Clonal analysis of high-grade squamous intra-epithelial lesions of the uterine cervix. AB - We previously reported that invasive squamous cell carcinomas of the uterine cervix are of monoclonal composition. In the current study, we extended our previous work to determine the clonal composition of cases of high-grade squamous intra-epithelial lesion (HSIL). Clonal analysis targeting the HUMARA locus was performed on cervical tissue from 9 cases, 8 showing heterozygosity at the HUMARA locus and being, therefore, informative for clonality analysis. Uterine cervices were cut into 12 blocks, fixed with formalin and embedded in paraffin, and DNA was extracted from targeted lesions of each block. A total of 30 samples of cervical intra-epithelial neoplasia 3 (CIN3) (14 samples of carcinoma in situ and 16 samples of severe dysplasia) and 1 sample of CIN2 (moderate dysplasia) were analyzed. Monoclonal composition of the lesions was demonstrated in 30/30 cases of CIN3. Polyclonal composition was seen in the single case of CIN2. In 6 uterine cervices, in which dysplastic lesions were present in more than 3 blocks, the pattern of X-chromosome inactivation was the same in all lesions, suggesting that these individual lesions were derived from a single cell, with intraepithelial extension within the cervical mucosa. By contrast, one uterus contained 2 discontinuous dysplastic foci with different patterns of X-chromosome inactivation, indicating that the 2 lesions developed independently from each other. Our results demonstrate that (i) lesions of CIN3 (severe dysplasia and carcinoma in situ) are composed of a clonal neoplastic population of cells and (ii) most cases of HSIL are unifocal in origin. PMID- 9359480 TI - Electrical power lines and childhood leukemia: a study from Greece. AB - Residential proximity to electrical power lines of different voltage in relation to childhood leukemia was investigated through a case-control study undertaken in Greece during 1993-1994. The study comprised 117 incident cases of childhood leukemia and 202 age-, gender- and place-of-residence-matched controls. Four measures of exposure to magnetic fields were developed, using data provided by the Public Power Corporation of Greece: Voltage (V) divided by the distance (d), V/d2, V/d3 and an adaptation of the Wertheimer-Leeper code. Conditional-logistic regression modeling was used to adjust for potential confounding influences of 18 variables. No significant trends of childhood leukemia risk with increasing exposure levels were noted, nor were there statistically significant elevations of disease risk at the higher exposure levels in each measure of exposure. These results do not support a causal link between residential proximity to electrical high-voltage wires and childhood leukemia risk, but in themselves do not refute a weak empirical association. PMID- 9359478 TI - Nasal T/natural killer (NK)-cell lymphomas are derived from Epstein-Barr virus infected cytotoxic lymphocytes of both NK- and T-cell lineage. AB - The cellular nature of nasal T/natural killer (NK)-cell lymphomas (NLs) remains controversial. It is still debatable whether these represent T-cell lymphomas with extensive loss of surface antigens or are, in fact, true NK-cell lymphomas. They are associated closely with Epstein-Barr virus (EBV), to the extent that EBV encoded small non-polyadenylated RNAs (EBER) expression can be used as a marker for the neoplastic cells. The cell lineage of this group of lymphomas was examined further by correlating immunophenotype, genotype and EBV status with the expression of cytotoxic granule-associated proteins, perforin and T-cell intracellular antigen-1 (TIA-1) in 13 cases of NL. Combined immunophenotypic and gene rearrangement analyses demonstrated that NLs can be identified clearly as either NK-cell or T-cell tumours. Nasal NK-cell lymphomas lacked clonal rearrangement of both T-cell receptor (TCR) gamma and immunogloulin heavy chain (IgH) genes and were either CD3(Leu4)-CD56+ (8 cases) or CD3(Leu4)+CD56+ (2 cases), whereas nasal T-cell lymphomas had rearranged TCRgamma and germ-line IgH genes and were either CD3(Leu4)+CD56+ (2 cases) or CD3(Leu4)+CD56- (1 case). Immunohistochemical (IH) studies showed that both perforin and TIA-1 were expressed universally in NL, irrespective of NK- or T-cell lineage. Dual labelling of TIA-1 by IH and EBER by in situ hybridisation demonstrated that the granule proteins were expressed predominantly by the EBER+ tumour cells. Our results indicate that NLs are derived from EBV-infected cytotoxic lymphocytes of both NK- and T-cell lineage. We postulate that cytotoxic lymphocytes generated during the cellular immune response to EBV infection or re-activation at the nasal region themselves may become targets for EBV infection and subsequent transformation. PMID- 9359481 TI - Ethnicity and variation in breast cancer incidence. AB - A breast cancer case-control study in Atlanta and 5 counties of central New Jersey involving interviews with 960 white and 281 black cases younger than 54 years of age enabled assessment of reasons for the varying incidence rates among these 2 ethnic groups. Of interest was why rates of breast cancer are higher among older white women, a trend that is reversed among very young women (<40 years). Calculation of the prevalence of exposure to classic and speculative risk factors and associated relative risks enabled derivation of population attributable risks (PARs) for the various combinations of age and ethnic groups. A higher PAR was derived for older (40-54 years) white (62%) than black (54%) women, which appeared to account for the observed difference in incidence between the 2 ethnic groups. Most of the difference in PARs between older whites and blacks was accounted for by whites having fewer births, later ages at first birth and slightly higher risks associated with reproductive and menstrual factors. Consideration of only well-established breast cancer risk factors showed a PAR among older whites of 57%, an estimate comparable to those previously published. Slightly higher overall PARs were derived when analyses considered several speculative but modifiable risk factors, including years of use of oral contraceptives, body size and alcohol consumption. Many of the analyses among younger women (20-39 years) were limited by available numbers, but it appeared that very little disease occurrence in young black women was associated with the factors studied. PMID- 9359482 TI - Prevailing papillomavirus types in non-melanoma carcinomas of the skin in renal allograft recipients. AB - The role of human papillomavirus (HPV) in the aetiology of in situ and invasive carcinoma of the genital tract is well established. In the rare disorder epidermodysplasia verruciformis (EV), in which patients develop extensive warts of unusual types and multiple cutaneous squamous cancers on light-exposed skin, current evidence suggests a probable role for a specific group of EV HPVs in the carcinogenic process. Determination of the possible role of HPV in the aetiology of non-melanoma skin cancers (NMSCs), which occur frequently in immunosuppressed organ allograft recipients, has been limited, until recently, by the lack of availability of a sensitive detection system for a wide range of cutaneous HPV types. We have used a combination of 2 sets of PCR primers to examine 68 benign and malignant tumours collected over a 12-year period from 25 renal allograft recipients. Cloning and sequencing of the PCR products were carried out to distinguish HPV DNA from cellular sequences. A combination of these techniques revealed HPV DNA in all viral warts, 65% of keratoses, 91% of intra-epidermal cancers and 91% of invasive squamous cancers. Both cutaneous and EV HPV types were detected, including 18 novel types. In 4 patients with multiple cancers, the most prevalent types were in the EV group: HPV 20, 23, 38 and 2 novel types, DL40 and DL267 (related to HPV 10 and 38, respectively). These 5 HPV types were present in a total of 73% of all malignant lesions tested. The technique described represents a reliable method of HPV DNA detection in NMSC. The EV group of HPVs predominate in the cancers, but the multiplicity of HPV types detected with double infection in some lesions suggests virus/virus in addition to virus/host interaction in the carcinogenic process. PMID- 9359483 TI - Mechanisms for high methoxymorpholino doxorubicin cytotoxicity in doxorubicin resistant tumor cell lines. AB - Methoxymorpholino doxorubicin (MMRDX) is an anthracycline analogue that is able to overcome tumor cell resistance to classical anthracyclines. Mechanisms for increased MMRDX cytotoxicity were analyzed in a small cell lung carcinoma cell line (GLC4), its 300-fold doxorubicin-resistant and multidrug resistance associated protein (MRP)-over-expressing subline (GLC4/ADR), an ovarian carcinoma cell line (A2780) and its 100-fold doxorubicin resistant and P-glycoprotein (P gp)-overexpressing subline A2780AD. Cross-resistance, measured with the MTT assay at MMRDX concentration resulting in 50% growth inhibition, was 1.8-fold in GLC4/ADR and 4.5-fold in A2780AD compared to their respective parental cell lines. Cellular MMRDX accumulation was equal in GLC4 and GLC4/ADR and 2-fold lower in A2780AD compared to A2780. Doxorubicin fluorescence was analyzed with confocal laser scan microscopy. Fluorescence was nuclear in sensitive, and cytoplasmic in resistant, cell lines, while MMRDX fluorescence was found in the nucleus in all cell lines. Pre-incubation with the MRP blocker MK 571 restored in GLC4/ADR cells the nuclear doxorubicin fluorescence pattern, as observed in GLC4 cells. MMRDX, thus, can largely overcome cross-resistance in these P-gp- and MRP overexpressing doxorubicin-resistant cell lines. Our results suggest that MMRDX is not a substrate for MRP-mediated resistance. PMID- 9359484 TI - Novel combination therapy for human colon cancer with adenovirus-mediated wild type p53 gene transfer and DNA-damaging chemotherapeutic agent. AB - Alteration of the wild-type (wt) p53 gene by mutation, deletion or re-arrangement is a major factor in the development of human colon cancer. Recent studies have demonstrated that p53 might be an essential component of the apoptotic pathway triggered by DNA-damaging stimuli such as chemotherapeutic agents and ionizing radiation. We examined the anti-tumor effects of adenovirus-mediated wt-p53 gene transfer in combination with a chemotherapeutic drug on the human colon cancer cell line WiDr, which is homozygous for a mutation in the p53 gene. Treatment with the chemotherapeutic drug cisplatin following infection with a replication deficient, recombinant adenoviral vector expressing wt-p53 (termed AdCMVp53) significantly suppressed the growth of WiDr cells compared to single treatments alone. To evaluate the in vivo efficacy of AdCMVp53 and cisplatin given sequentially, WiDr cells were inoculated s.c. in nu/nu mice. After 3 days, AdCMVp53 was injected s.c. into the area where tumor cells were implanted, followed by i.p. administration of cisplatin. Analysis of initial growth inhibition at 21 days demonstrated a profound therapeutic cooperativity, though administration of either AdCMVp53 or cisplatin alone was followed only by a slowing of growth. Our results suggest that gene therapy using wt-p53-expressing adenovirus in combination with a chemotherapeutic DNA-damaging drug could be a useful strategy for treating human colon cancer. PMID- 9359485 TI - Effects of chronic indomethacin therapy on the development and progression of spontaneous mammary tumors in C3H/HEJ mice. AB - The present study was designed to test the hypothesis that endogenous prostaglandin E (PGE) promotes the development, growth and metastasis of spontaneous mammary tumors in C3H/HeJ female retired breeder mice. The effect of chronic oral indomethacin (indo) therapy starting at 6 months of age was tested on these parameters as well as on animal survival, in comparison with control mice placed on 0.2% ethanol in drinking water for up to 25 months of age. Indo treatment delayed the initial (up to 27 weeks) development of primary tumors by 11-12 weeks; however, the subsequent rate of tumor appearance was unaffected (totaling 82% in indo-treated vs. 90% in controls by 25 months of age). Spontaneous regression of primary tumors (26% in controls) increased 2-fold (53%) with indo therapy. While the apparent reduction in the growth rate of primary tumors and the overall prolongation of animal survival were not significant, the lifespan of mice bearing multiple tumors was significantly prolonged by therapy. There was also a 2-fold reduction in the incidence of lung metastases in mice bearing detectable primary tumors, and this was more pronounced during the earlier phase of tumor development. Positive immunostaining for cyclooxygenase-2 enzyme (indicative of the cellular source of PGE) was exhibited by tumor cells, stromal cells and macrophages within the primary tumors. Tumors in indo-treated mice exhibited histological evidence of increased differentiation (acinar architecture), significant tumor cell death, mononuclear cell infiltration and reduction in vascularity, indicating that the beneficial effects of indo were due to multiple mechanisms, including improved immune response and reduced angiogenesis. PMID- 9359486 TI - Comparative effects of 28-day treatment with the new anti-estrogen EM-800 and tamoxifen on estrogen-sensitive parameters in intact mice. AB - Following 28 days of oral administration, in intact mice, the novel non-steroidal anti-estrogen EM-800 was at least 30-fold more potent than tamoxifen in inhibiting uterine weight. Moreover, the maximal inhibitory effect achieved with tamoxifen on uterine weight was only 40% that with EM-800. The pure anti estrogenic activity of EM-800 on the hypothalamo-pituitary-gonadal axis is illustrated by the increase in ovarian weight, while tamoxifen, due to its estrogenic activity, decreased ovarian weight. EM-800 is 10- to 30-fold more potent than tamoxifen in inhibiting uterine and vaginal estrogen receptors. Since 17beta-hydroxysteroid dehydrogenase (17beta-HSD) is the key enzyme in estradiol formation, the potent inhibitory effect of EM-800 on uterine 17beta-HSD could play an additional role by decreasing the availability of estradiol in the uterine tissue, while tamoxifen, on the contrary, stimulates activity of the enzyme. The atrophic changes in both the endometrial and myometrial layers achieved with EM-800 almost reached those observed 28 days after ovariectomy. EM 800 also resulted in a marked decrease in the number of ovarian developing follicles and corpora lutea, while the number of atretic follicles was increased. Tamoxifen treatment, on the other hand, produced an increase in both the number and crowding of the endometrial glands and a mild atrophy of the myometrial layer. Tamoxifen caused atrophic changes of the vaginal epithelium, especially at the highest doses, though the atrophy was much less pronounced than that following EM-800 treatment or ovariectomy. In addition to being at least 30-fold more potent than tamoxifen in inhibiting uterine weight, the novel anti-estrogen causes atrophy of the endometrium, stimulates the hypothalamo-pituitary-gonadal axis and inhibits uterine 17beta-HSD activity, while tamoxifen exerts opposite and estrogen-like effects on these parameters. PMID- 9359487 TI - Cure of malignant ascites and generation of protective immunity by monoclonal antibody-targeted activation of a glucuronide prodrug in rats. AB - We examined the in vivo efficacy of targeting beta-glucuronidase (betaG) to activate a glucuronide prodrug (BHAMG) of p-hydroxyaniline mustard (pHAM) at hepatoma ascites in Sprague-Dawley rats. Injection i.p. of 500 microg RH1-betaG, a conjugate formed between recombinant betaG and monoclonal antibody RH1 with specificity for an antigen expressed on AS-30D rat hepatoma cells, into rats bearing AS-30D ascites resulted in the accumulation of 54 microg conjugate per 10(9) tumor cells after 2 hr. Ascites fluid and serum contained 0.53 and 0 microg/ml, respectively, RH1-betaG 2 hr after injection of the conjugate. Conjugate binding to AS-30D cells was heterogeneous and non-saturated, as determined by flow cytometry. BHAMG was less toxic than pHAM to SD rats based on measures of animal mortality, weight loss and hematological toxicity. Treatment of rats bearing established hepatoma ascites with 500 microg RH1-betaG followed 2 hr later with a single i.p. injection of 30 mg/kg BHAMG or 3 i.p. injections of 10 mg/kg BHAMG 2, 3 and 4 hr later resulted in the cure of 6/8 and 8/8 animals, respectively. Treatment with BHAMG or pHAM alone did not produce cures, whereas treatment with a control antibody-betaG conjugate and BHAMG produced significantly greater hematological toxicity compared to treatment with RH1-betaG and BHAMG. All cured rats were completely protected from rechallenge with 2 x 10(7) AS-30D cells, indicating that successful treatment of animals induced protective immunity. PMID- 9359488 TI - Analysis of the interaction of procathepsin D activation peptide with breast cancer cells. AB - Cathepsin D, a lysosomal aspartic proteinase, is secreted in the form of enzymatically inactive proenzyme by many types of human breast cancer tissue and exerts mitogenic activity toward these tissues. Flow cytometry was used to test the binding of procathepsin D purified from the secretion of the breast cancer cell line ZR-75-1 to human breast cancer cells. No previously known surface antigens or soluble M6P-R or anti-M6P-R antibodies were found to inhibit the specific binding of procathepsin D-FITC. Similarly, none of these potential inhibitors was found to inhibit growth factor activity of procathepsin D. Our results indicate that procathepsin D growth factor activity is mediated by a new, previously unknown receptor moiety and that the binding activity can be localized in position 27-44 of the activation peptide of procathepsin D. Furthermore, in vivo experiments indicate that treatment with anti-procathepsin D antibodies can reverse the growth of human breast tumors in athymic nude mice. PMID- 9359489 TI - Suppression of pulmonary metastasis in murine B16 melanoma cells by transfection of a sialidase cDNA. AB - A cytosolic sialidase cDNA was transfected into a highly metastatic and invasive cell line, B16-BL6, derived from the murine B16 melanoma. Stable transfection of a cytosolic sialidase expression vector yielded 4 transfectants with high content of the exogenous sialidase protein as well as enzyme activity. These transfectants exhibited markedly decreased experimental pulmonary metastasis, invasiveness in collagen gels and cell motility on colloidal gold-coated glass plates but no change in cell attachment to fibronectin, collagen type VI or laminin. To cast light on the underlying mechanisms, cellular constituents of the transfectants were analyzed. Sialidase over-expression did not lead to any significant changes in cell surface carbohydrates or intracellular glycoproteins, as revealed by lectin flow cytometry and lectin blotting, respectively. Thin layer chromatography of intracellular glycolipids, however, revealed decreased ganglioside GM3 and increased lactosylceramide as major changes. PMID- 9359490 TI - Treatment of the P815 murine mastocytoma with cisplatin or etoposide up-regulates cell-surface Fas (CD95) expression and increases sensitivity to anti-Fas antibody mediated cytotoxicity and to lysis by anti-CD3-activated killer-T cells. AB - We have investigated the effect of pre-treatment with the anti-cancer drugs cisplatin and etoposide on the susceptibility of P815 murine mastocytoma cells to lysis by murine spleen-derived anti-CD3-activated killer-T (AK-T) cells. A 20 hr pre-treatment with cisplatin (0.2-2 microg/ml) or etoposide (0.01-1 microg/ml) rendered P815 cells significantly more sensitive to AK-T cell-mediated lysis in a 4 hr 51Cr-release assay than untreated control tumor cells. At lower concentrations, pre-treatment with cisplatin or etoposide had no direct cytotoxic effects on P815 tumor cells, as measured by the MTT assay. AK-T cell-mediated killing of P815 tumor cells pre-treated with 2 microg/ml cisplatin or 1 microg/ml etoposide was only partially inhibitable by the Ca2+ chelator EGTA, suggesting that the Ca2+-independent Fas (CD95)/Fas ligand cytolytic pathway of AK-T cells contributes to cytotoxicity. In comparison to untreated control P815 cells, 2 microg/ml cisplatin- or 1 microg/ml etoposide-treated P815 cells exhibited increased expression of Fas mRNA and cell-surface Fas, which correlated with increased sensitivity to lysis by AK-T cells. In addition, pre-treatment with cisplatin or etoposide caused P815 tumor cells to become sensitive to the cytotoxic effects of anti-Fas antibody in a 4 hr 51Cr-release assay. Taken together, our results demonstrate that short-term exposure to concentrations of cisplatin and etoposide in the low cytotoxic range and below up-regulates Fas expression by P815 tumor cells, thereby facilitating cytotoxicity mediated through the Fas/Fas ligand cytolytic pathway. PMID- 9359491 TI - Stimulation of proliferation in human colon cancer cells by human monoclonal antibodies against the TF antigen (galactose beta1-3 N-acetyl-galactosamine). AB - In many tissues, the TF (Thomsen-Friedenreich) blood group antigen (Galbeta1 3GalNAc alpha-) behaves as an onco-foetal carbohydrate antigen, showing increased expression in malignancy and hyperplasia. Dietary lectins which bind the TF antigen have marked effects on proliferation of epithelial cells without cytotoxicity. This led us to speculate that anti-TF antibodies, including those that naturally occur in humans, might have similar effects. Five anti-TF antibodies, TF2 (human), TF5 (human), 5A8 (mouse), 8D8 (mouse) and BM22 (mouse), but not TFI (human) or 49H.9 (mouse), showed marked dose-dependent stimulation (95-192%) of [3H]thymidine incorporation by HT29 human colon cancer cells. Similar stimulation of proliferation of HT29 cells by these monoclonal antibodies (MAbs) was found when cell count assessment was used. Antibody-stimulated proliferation was inhibited by co-incubation with glycoproteins expressing Galbeta1-3GalNAc alpha- (asialo glycophorin or [Galbeta1-3GalNAc alpha-O-p aminophenyl]n-human serum albumin). A proliferative effect of these antibodies was also demonstrated on human colon cancer cell lines LS174T and HT29-MTX but not on Caco-2 cells. Although immunoblotting showed similar binding patterns of all the antibodies on HT29 cell membrane extracts, there was little correlation between cell surface binding assessed by immunofluorescence and proliferative response, and internalization of the biotinylated antibody TF5 was demonstrated by confocal microscopy. Our results provide further evidence that cell surface glycoproteins which express TF antigen may play an important role in the regulation of cell proliferation and also suggest that human anti-TF antibodies may have proliferative effects on cells which express TF antigen. PMID- 9359492 TI - Human squamous-cell-carcinoma cell line (DJM-1) cells synthesize P-cadherin molecules via an elevation of extracellular calcium: calcium regulates P-cadherin gene expression at the translational level via protein tyrosine phosphorylation. AB - Spatially-regulated P-cadherin expression is crucial for maintaining the normal epidermal architecture. P-cadherin expression in cutaneous squamous-cell carcinomas (SCC) is altered, and may participate in tumor progression. We therefore investigated how P-cadherin expression was regulated in a cultured cutaneous SCC cell line (DJM-1). At low calcium concentration (0.05 mM), DJM-1 cells expressed P-cadherin weakly in the cytoplasm. At a higher calcium concentration, P-cadherin was promptly translocated to the cell surface within 30 min, gradually increased on the cell surface for up to 48 hr, and was continuously expressed for at least 7 days. During this time course, the total amount of P-cadherin protein had increased, whereas the steady-state mRNA levels for P-cadherin had not changed. The inhibition of protein synthesis by cycloheximide, but not the inhibition of gene transcription by actinomycin-D, completely suppressed the expression of P-cadherin. The effect of calcium was inhibited by tyrphostins but not by H-7, cholera toxin, or dibutylic cyclic AMP. Increments in the extracellular calcium concentration did not mobilize the intracellular calcium pool, and were accompanied by the tyrosine phosphorylation of a 62-kDa protein. In addition, DJM-1 cells expressed mRNA for a calcium sensing receptor originally demonstrated in the parathyroid gland. The results suggest an unique mechanism for regulating P-cadherin gene expression in DJM-1 cells by extracellular calcium, which stimulates the de novo synthesis of P cadherin at the translational level through protein tyrosine phosphorylation. PMID- 9359493 TI - Induction of apoptosis by cryptophycin 1, a new antimicrotubule agent. AB - The ability of cryptophycin 1, a new potent cytotoxic antimicrotubule agent, to initiate apoptosis was studied. Treatment of cells with cryptophycin 1 (50 pM) rapidly caused morphological changes consistent with the induction of apoptosis. DNA strand breakage and fragmentation of the DNA into oligonucleosome-sized fragments was observed, and this coincided with the loss of cellular DNA. Activation of the cysteine protease CPP32 (caspase 3, YAMA, apopain), a member of the ICE/CED-3-like protease family of apoptosis effectors, was consistent with the execution of cell death by a coordinated sequence of events. Low concentrations of cryptophycin 1 caused mitotic arrest with the formation of abnormal mitotic spindles without affecting interphase microtubule structures. Unlike other microtubule active agents, cryptophycin-induced mitotic arrest persisted for only a brief period before the onset of apoptosis. There was no evidence of release from G2/M cell cycle arrest. Our results show that low concentrations of cryptophycin 1 (50 pM) initiated cell death consistent with apoptosis. These data suggest that the cytotoxic effects of cryptophycin 1 are due in part to its ability to initiate apoptosis rapidly. PMID- 9359494 TI - Introduction of p53 induces cell-cycle arrest in p53-deficient human medullary thyroid-carcinoma cells. AB - The structural integrity of the p53 gene in a human thyroid-medullary-carcinoma derived cell line has been studied. Analysis of high-molecular-weight DNA showed that the p53 locus is severely rearranged. PCR and single-strand conformation polymorphism analysis revealed that a large portion of the 5' end of the p53 gene is lost, while a region encompassing exons 8 and 9 is rearranged. As a consequence, no virtual expression of a p53-specific transcript is detected in mRNA from the medullary-carcinoma cell line. The absence of a p53 protein prompted us to analyze the biological effect of exogenous expression of this tumor-suppressor gene on cell growth and viability, introducing retroviral constructs carrying full-length human wild-type p53 cDNA. Contrary to what has been described for other cell types, including most thyroid-carcinoma cell lines of follicular origin, these experiments allowed us to establish clonal-cell populations which constitutively express p53. Cytometric analysis revealed G1 specific cell-cycle arrest, responsible for growth retardation in the transfected clones when compared with the parental cell line. However, medullary-thyroid carcinoma cells expressing p53 are able to partially overcome the G1 block and progress through the cell cycle. In the search of the mechanism(s) involved in these processes, we describe the interaction of p53 with specific p21WAF1/Cip1 promoter sequences by gel-retardation assays. PMID- 9359495 TI - The haematological features of HIV infection. PMID- 9359496 TI - Haemopoietic stem cell transplantation for autoimmune diseases. PMID- 9359497 TI - Survival of patients with chronic myelogenous leukaemia relapsing after bone marrow transplantation: comparison with patients receiving conventional chemotherapy. AB - Treatment with busulphan and/or hydroxyurea rarely produces remission in patients with chronic myelogenous leukaemia (CML) in chronic phase. HLA-identical sibling transplants almost always produce remission, and only about 20% of patients relapse post-transplant. The increased anti-leukaemic efficacy of transplants results from intensive pretransplant treatment and immune-mediated anti-leukaemia effects. We studied 433 patients surviving > or = 2 years after diagnosis of CML to determine if patients who have relapsed after a transplant in chronic phase have longer survival from diagnosis than comparable subjects receiving chemotherapy. The chemotherapy cohort included 344 adults < 50 years of age treated on consecutive trials of the Italian Cooperative Study Group on CML between 1973 and 1986. The transplant cohort included 89 patients reported to the International Bone Marrow Transplant Registry who relapsed after an HLA-identical sibling bone marrow transplant carried out between 1978 and 1992. Survivals in the two groups were compared using Cox proportional hazards regression to adjust for prognostic variables. Median survival was 65 months in the chemotherapy cohort and 86 months in the transplant cohort. The 7-year probability (95% confidence interval) of survival was 34% (28-39%) in the chemotherapy cohort and 57% (43-70%) in the transplant cohort (P=0003). There was no difference in survival of patients relapsing after T-cell depleted and non-T-cell-depleted transplants. We conclude that patients who relapse after an HLA-identical sibling bone marrow transplant for CML in chronic phase have longer survival from diagnosis than comparable patients receiving chemotherapy. This effect is most likely to be the result of intensive chemotherapy and/or radiation given for pretransplant conditioning. PMID- 9359498 TI - Bone marrow transplantation for chronic myeloid leukaemia: the effects of differing criteria for defining chronic phase on probabilities of survival and relapse. AB - We studied actuarial survival and relapse in 251 patients with chronic myeloid leukaemia (CML) treated by bone marrow transplantation (BMT) from HLA-identical sibling donors at a single institution. According to the institutional criteria used to define disease phase at the time of BMT. the 5-year probabilities of survival were 58.1% (95% confidence interval 50-66%) for 205 chronic-phase patients and 21.5% (95%CI 12-37%) for 46 advanced-phase patients (P<0.00001); the corresponding values for relapse were 34.8% (95%CI 27-44%) versus 72.7% (95%CI 46 89%). When disease phase was defined strictly according to the criteria of the International Bone Marrow Transplant Registry, the survival for 154 chronic-phase patients increased to 60.1% (95%CI 51-69%) and that for 97 advanced-phase patients increased to 37.6% (95%CI 28-48%). There was a parallel change in probabilities of relapse in the two patient groups (33.9% [95%CI 25-44%] and 51.3% [95%CI 37-66%], respectively). We also observed that patients transplanted in advanced phase had a higher incidence of grades III-IV acute graft-versus-host disease (P=0.001) and transplant-related mortality (P=0.02) than those undergoing BMT for chronic-phase disease. We recommend that transplant centres reporting results of BMT should always specify the precise criteria used for defining disease phase in order to ensure that results between different centres are strictly comparable. PMID- 9359499 TI - Unrelated donor bone marrow transplantation in children and young adults with acute myeloid leukaemia in remission. AB - The role of unrelated donor bone marrow transplantation (UD-BMT) in the management of patients with acute myeloid leukaemia (AML) is uncertain. We describe 18 patients with a median age of 13 years (range 4-31) who received an ex vivo T-cell-depleted UD-BMT for AML (13 in second complete remission (CR2) and five in first complete remission (CR1) with high-risk features). Nine donor recipient pairs were fully matched; eight of these donor-recipient pairs had a single class I HLA mismatch; one patient had both single class I and class II HLA mismatches. Grade II GVHD of the skin occurred in four patients (22%) and limited chronic GVHD in two patients (11%). There have been four deaths: one from relapse and three from infection. With a median follow-up of 27 months, 14 patients survive and the actuarial event-free survival at 2 years is 70 +/- 20% (95% confidence interval). We conclude that unrelated donor BMT can result in prolonged disease-free survival in children and young adults with AML. PMID- 9359500 TI - Peripheral blood progenitor cell mobilization in patients with primary refractory lymphoma or at first relapse: comparison with patients at diagnosis and impact on clinical outcome. AB - Peripheral blood progenitor cell (PBPC) mobilization was evaluated in 53 patients receiving the high-dose sequential (HDS) regimen: 27 had non-Hodgkin's lymphoma or Hodgkin's disease, primary refractory or at first relapse, 26 had non Hodgkin's lymphoma at diagnosis. Mobilization was assessed following either 7 g/m2 cyclophosphamide (48 patients) or 2 g/m2 etoposide, both followed by G-CSF (filgrastim) at 5 microg/kg/d. PBPC mobilization was significantly higher in patients at diagnosis compared to refractory/relapsed patients (median peak values of circulating CFU-GM: 25,209/ml v 4270/ml, P < 0.0001 and CD34+ cells: 286/microl v 47/microl, P < 0.0001). All patients receiving HDS as up-front treatment mobilized enough PBPC for an autograft, often requiring a single leukapheresis; whereas only 15 patients under salvage treatment with HDS were able to complete PBPC autograft. Bone marrow (BM) cells, alone or with PBPC, were needed in six patients, and autograft could not be performed in six patients. Among refractory/relapsed patients, those having a high PBPC mobilization experienced a significantly longer EFS compared to those who had not; autograft completion also significantly enhanced EFS. Thus, the use of an effective mobilizing protocol does not ensure adequate PBPC mobilization in moderately pretreated patients; low mobilization must be considered as an early sign of poor outcome in patients receiving a high-dose salvage programme. PMID- 9359501 TI - Separation of G-CSF-mobilized PBSC transplants by counterflow centrifugal elutriation: modest enrichment of CD34+ cells but no loss of primitive haemopoietic progenitors. AB - The suitability of counterflow centrifugal elutriation (CCE) for reduction of the number of non-stem cells in autologous G-CSF-mobilized peripheral blood stem cell (PBSC) transplants was investigated. By cell size-monitored CCE, small cells could be rapidly separated from the haemopoietic progenitor cells present in leukapheresis product (LP) samples. The large cell fraction contained an average 86 +/- 25% of the CD34+ cells and 76 +/- 20% of the granulocyte-macrophage progenitors (CFU-GM) loaded into the separation chamber, and was depleted of 75 +/- 18% of the lymphocytes, 89 +/- 7% of the erythrocytes and 98 +/- 2% of the platelets (n = 21). Due to the presence of high numbers of large immature myeloid cells, which co-elutriated with progenitor cells, enrichment of CD34+ cells in the large cell fraction was only modest (average 1.8 times). No indication of preferential co-elutriation of primitive stem cells with the small cells was obtained. There was no difference in expression of CD38 or Thy-1 on CD34+ cells between the two elutriation fractions. Frequencies of cobblestone-area-forming cells (CAFC) week 6, which are considered to represent cells with long-term repopulating ability, were reduced in the small cell fractions as compared to those in the unseparated samples and the large cell fractions. On average, 100% of CAFC week 6 were recovered in the large cell fractions (n = 5). In conclusion erythrocytes, platelets and 40-50% of leucocytes can be depleted from G-CSF mobilized PBSC samples by CCE with an almost complete recovery of both clonogenic and primitive stem cells. PMID- 9359502 TI - Autologous peripheral blood stem cell transplantation in a patient with chronic autoimmune thrombocytopenia. AB - Immunoablation by high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (PBSCT) is one approach currently under discussion for the treatment and possible cure of patients with severe autoimmune diseases. Chronic immune thrombocytopenic purpura (ITP) can be refractory to current therapies and in severe cases may constitute a life-threatening condition. We performed autologous PBSCT with T-cell depletion in a patient with severe chronic ITP but observed no effect on platelet levels. This contrasts with a recent report where complete remission was induced by a similar regimen in two patients with ITP Further studies will be necessary to identify subgroups of patients who might benefit from this form of therapy. PMID- 9359503 TI - Sibling allogeneic bone marrow transplantation in a patient with type I Glanzmann's thrombasthenia. AB - Glanzmann's thrombasthenia is a rare inherited bleeding disorder caused by either quantitative or qualitative abnormalities of the platelet membrane glycoprotein (Gp) IIb/IIIa complex. Bleeding is usually mucocutaneous in origin and may be of a severe nature. We report the use of allegeneic bone marrow transplantation in a 5-year-old child with homozygous type I Glanzmann's thrombasthenia, using the patient's younger brother as marrow donor. Engraftment was successful and has resulted in a resolution of bleeding episodes. We conclude that allogeneic BMT is a potentially curative option for those with Glanzmann's thrombasthenia associated with severe bleeding symptoms. PMID- 9359504 TI - Disparate lympho-erythroid donor to recipient chimaerism in a beta(0) thalassaemia bone marrow transplant recipient with red cell indices indicative of apparent full engraftment. AB - A 4-year-old girl with transfusion-dependent beta(0)-thalassaemia received an HLA identical bone marrow transplant (BMT) from her beta(0)-thalassaemia trait sister. Prior to BMT, chromosomal analysis revealed the recipient to have 46,XX,9qh+, a polymorphic variant of the heterochromatin region of chromosome 9, which her donor did not have. Within 1 month post-BMT, 89% of nucleated bone marrow cells were of donor origin. One year later, donor engraftment had decreased to 44% and 34% in nucleated bone marrow cells and blood lymphocytes, respectively. By 2 years, donor lymphocyte engraftment fell to 5%, raising concern of possible graft rejection. To examine erythroid chimaerism, globin synthesis by individual erythroid progenitor cell derived colonies (BFU-E) was analysed. On days 1000 and 1130 post-BMT, 79% and 77% of colonies, respectively, synthesized beta-globin and therefore were of donor origin. PMID- 9359505 TI - Minimal residual disease in acute myelogenous leukaemia and myelodysplastic syndromes: a follow-up of patients in clinical remission. AB - The majority of patients with acute myelogenous leukaemia (AML) and myelodysplastic syndromes (MDS) relapse, especially those with unfavourable cytogenetics. This study was designed to investigate the presence and frequency of minimal residual disease (MRD) in patients with AML or MDS (n=35) and numerical abnormalities of chromosomes 6, 7, 8, 9, 10, 17 and 18 in clinical remission by using a combination of fluorescence activated cell sorting (FACS), fluorescence in-situ hybridization (FISH) and labelling with bromodeoxyuridine (BUdR). The technique enables the detection of as few as three leukaemic cells in 10(5) normal cells. MRD was detected in 33/35 patients in complete remission (CR). 16 patients relapsed (8/11 with monosomy 7, 4/17 with trisomy 8, and 4/7 with other cytogenetic abnormalities) after a median of 4.8 months (range 3-13). Levels of MRD (P=0.007) and proliferation index (P=0.011) were significantly higher in patients with monosomy 7 than in patients with trisomy 8 or other cytogenetic abnormalities. The percentage of cells in S-phase, the number of abnormal cells and cytogenetic class were related to time to relapse (P=0.001) with S-phase being the single most important prognostic factor (P=0.0001). We conclude that the combination of FACS/FISH/BUdR, which determines the number, phenotype and proliferation rate of very rare leukaemic cells in patients with AML or MDS in clinical remission, provides information that is useful in the identification of patients with high and low likelihood of relapse. PMID- 9359506 TI - MDR 1 expression is an independent prognostic factor for response and survival in de novo acute myeloid leukaemia. AB - The Multidrug Resistance gene (MDR 1) is frequently expressed in acute myeloid leukaemia (AML). MDR 1 is associated with resistance to chemotherapy in vitro and with a poor response rate in AML. We have investigated the prognostic value of MDR 1 expression in relation to other patient characteristics with respect to response and survival. One hundred and thirty patients aged 0-88 years were treated for de novo AML with standard induction and consolidation chemotherapy. MDR 1 expression was determined by immunocytochemistry. Univariate and multivariate analyses were conducted to identify prognostic factors for reaching complete remission (CR) and for overall survival from diagnosis, in order to compare MDR 1 with known prognostic factors. Univariate analysis showed that higher MDR 1 expression was an adverse prognostic factor for CR (P<0.001), as was higher age (P<0.001) and unfavourable karyotype (P<0.01). These factors were also negative prognostic factors for overall survival (P<0.001, P<0.05 and P<0.005, respectively). In the multivariate analysis MDR 1 (P<0.001), higher age (P<0.001) and karyotype (P<0.01) were independent adverse prognostic factors for CR as well as for overall survival (P<0.001, P<0.005, P<0.001, respectively). Our data indicate that MDR 1 expression is a disease-related unfavourable prognostic factor which has a significant impact on complete remission and overall survival in AML. Analysis of MDR 1 may be used to determine prognosis in individual patients. PMID- 9359507 TI - Intensification of treatment for adults with acute lymphoblastic leukaemia: results of U.K. Medical Research Council randomized trial UKALL XA. Medical Research Council Working Party on Leukaemia in Adults. AB - The MRC UKALL XA trial for patients aged 15 years and over with acute lymphoblastic leukaemia was designed to evaluate short blocks of intensive 'AML style' treatment. Between 1985 and 1992, 618 eligible patients were entered into the trial. 450 patients were randomized to receive early intensification at 5 weeks, late intensification at 20 weeks, both, or neither. Unlike the concurrent children's trial, UKALL X, which was of similar design, UKALL XA does not demonstrate a clear benefit for intensification, although there was a significant reduction in the relapse risk due to the early block. The estimated increase in disease-free survival at 5 years was 2% with 95% confidence interval from 1% reduction to 5% increase. There may be a real difference between the effect of these treatments in adults and in children, but this result may be somewhat weakened by poorer compliance, with a greater proportion of adults not receiving the treatment arm to which they were randomized. PMID- 9359510 TI - Frequent deletion of chromosome 12p12.3 in children with acute lymphoblastic leukaemia. AB - Cytogenetic deletions of the short arm of chromosome 12 are common recurring alterations found in a wide range of haematological neoplasias, including childhood acute lymphoblastic leukaemia (ALL), the most frequent paediatric malignancy. Such a loss of genetic material suggests the presence of a tumour suppressor gene which plays an important role in growth regulation or in the differentiation of haemopoietic stem cells. To substantiate this hypothesis and to determine more precisely the chromosomal location of this putative gene, we applied a deletion mapping strategy based on the detection of loss of heterozygosity (LOH) at specific genomic loci in tumour cells. 13 polymorphic markers were used to screen DNA samples from 20 children with ALL. LOHs at 12p12.3 were observed in almost 50% of informative B-cell precursor ALL patients analysed. This is one of the most frequent genetic alterations found in this disease. A common region of LOH was delimited by the markers D12S89 (distal) and D12S358 (proximal), separated by a genetic interval of approximately 3 cM. We refined the position of the putative 12p tumour suppressor gene to a physical interval of <1.3 Mb, a crucial step towards the identification of candidate genes. A yeast artificial chromosome clone contig that spans the entire critically deleted region includes two known genes: TEL, a member of the ets family of transcription factors, and p27KIP1, a cyclin-dependent kinase inhibitor. PMID- 9359509 TI - The majority of myeloid-antigen-positive (My+) childhood B-cell precursor acute lymphoblastic leukaemias express TEL-AML1 fusion transcripts. AB - The t(12:21) translocation fuses the TEL and AML1 genes and has been found in up to 28% of paediatric B-cell precursor acute lymphoblastic leukaemias (BCP-ALL). The AML1 gene is a transcription factor which regulates expression of several myeloid differentiation associated genes. A molecular analysis of TEL-AML1, E2A PBX1, MLL-AF4, BCR-ABL expression and an immunophenotypic study of CD13/CD33 myeloid antigen expression have been performed prospectively on tumour cells from 96 paediatric BCP-ALL patients. Percentages of CD13 or CD33 expressing leukaemic cells were found to be higher in TEL-AML1 positive cases (n = 22) than in TEL AML1 negative (n = 74) cases (P<0.001). In 22/96 cases (23%) >10% of neoplastic cells were found to express at least one of the two markers. In 14 of these cases (63%), TEL-AML1 expression was detected, whereas t(4;11), t(11;19) and t(9;22) translocations were found by molecular methods in only three cases (14%). In four cases (18%) no molecular marker was found. These data show that TEL-AML1 expression is significantly associated with myeloid antigen expression by leukaemic cells and suggests that the prognostic significance of myeloid antigen expression in paediatric ALLs should be re-evaluated in the light of molecular cytogenetic markers. PMID- 9359508 TI - Cytogenetics and prognosis in childhood lymphoblastic leukaemia: results of MRC UKALL X. Medical Research Council Working Party in Childhood Leukaemia. AB - We have analysed the prognostic influence of cytogenetic findings at diagnosis in a group of 502 children with acute lymphoblastic leukaemia (ALL), treated on MRC UKALL X, in whom clonal cytogenetic abnormalities were detected at diagnosis. Despite the overall improvement in outcome in children treated on this protocol compared with previous trials, some cytogenetically-defined groups were still associated with a poor outcome and ploidy retained some prognostic significance. Patients with high hyperdiploid ALL (39% of those with clonal abnormalities) had a favourable outcome with event free survival of 71% at 5 years. Those with near haploidy (1%), hypodiploidy (9%) and low hyperdiploidy (16.5%) had a relatively poor prognosis with event-free survival at 5 years of 17%, 42% and 49% respectively. Only two of 12 children with Ph-positive leukaemia are alive in remission and abnormalities of chromosome 11q23 were also associated with a high risk of treatment failure. In contrast, the t(1;19) was associated with improved event-free survival of 87.5% at 5 years. A number of other non-random abnormalities were identified with no clear prognostic significance. We conclude that identification of certain genetic changes remains important in the management of acute lymphoblastic leukaemia, although whether molecular diagnosis of clinically relevant abnormalities can now supplant cytogenetics in the clinical trials context remains to be determined. PMID- 9359511 TI - Crosslineage TCR delta rearrangements occur shortly after the DJ joinings of the IgH genes in childhood precursor B ALL and display age-specific characteristics. AB - In order to test the hypothesis that the most immature T-cell receptor (TCR) rearrangements occur after the DJ joining of the immunoglobulin heavy chain genes (IgH), we analysed the TCR Vdelta2-Ddelta3 rearrangements in precursor B-cell leukaemias (PBC ALL) from 25 children younger than 3 years at disease onset and found that most of the junctional regions had N nucleotides inserted. We then selected 14 of these PCB ALLs for DJH (DJ joining of the IgH) characterization. These joining regions showed homology-directed recombination and lack of N regions, indicating absence of terminal deoxynucleotidyl transferase (TdT) activity during their rearrangement. Most leukaemias with a DJH rearrangement without N region have no, or only one, nucleotide in the joining regions of their Vdelta2-Ddelta3 rearrangements. The N regions of the TCR delta rearrangements displayed 'age-specific' differences: in children younger than 3 years of age the N regions were shorter than in those older than 3 years, and the rearrangements frequently contained complete segments. We conclude that the Vdelta2-Ddelta3 rearrangement in childhood PCB ALLs is an early event following DJH rearrangement and that it occurs shortly before or after the first hit, leading to malignant transformation. PMID- 9359512 TI - Polymerase chain reaction on cerebrospinal fluid cells in the detection of leptomeningeal involvement by B-cell lymphoma and leukaemia: a novel strategy and its implications. AB - In the search of B-cell lymphoma/leukaemia dissemination to cerebrospinal fluid (CSF), we used the highly sensitive semi-nested PCR (snPCR) for the analysis of IgH gene rearrangements. This method detects a rearranged IgH gene from a single B lymphocyte which may or may not represent the neoplastic B-cell population. We therefore performed multiple snPCR (three to five) experiments on the same CSF sample, postulating that the detection of a band of the same size and sequence in different PCR runs was highly indicative of a clonal population. 17 consecutive cases with a differential diagnosis of primary (PCNSL) (n=10) or secondary (SCNSL) (n=7) CNS lymphoma or leukaemia were investigated by the new strategy. The clonal nature of the B-cell population was confirmed in 3/10 of suspected PCNSL, and in six other cases the PCR study was indicative of reactive lymphocytosis. One case revealed a clonal B-cell population in the clinical context of an autoimmune disorder. Evidence of clonal B-cell population was found in 4/7 of suspected SCNSL. In one of these cases the detected band and its sequence proved identical to that of the primary nodal lymphoma. We believe that the evaluation of B-cell clonality in CSF requires multiple snPCR amplification on the same sample to compare the size of the products and, if necessary, the DNA sequences to ascertain the diagnosis of malignancy in equivocal cytologic and clinical findings. PMID- 9359513 TI - Cyclin D1 and p16INK4A are preferentially expressed in immature and mature myeloma cells, respectively. AB - Myeloma cells consist of immature, intermediate and mature cells with respect to expression of VLA-5 (CD49e) and MPC-1 adhesion molecules. VLA-5(-)MPC-1(-) immature myeloma cells respond to interleukin 6 (IL-6) to proliferate in vitro. but VLA-5+MPC-1+ mature myeloma cells have almost no proliferative activity with higher secretory activity of M-protein in vitro. In order to further clarify the biological differences between these immature and mature myeloma cells, we examined survival of these cells with or without IL-6 in vitro, and investigated the underlying mechanism of the proliferative or non-proliferative character of these cells by examining expression of cell cycle regulators such as cyclin D1 and inhibitors for cyclin-dependent kinase (Cdk), p16INK4A, p21CIP1 and p27KIP1 by RT-PCR and immunohistochemistry. In vitro survival of these myeloma cells was examined by flow cytometric quantification of fluorescein diacetate (FDA) and propidium iodide (PI) staining. Immature myeloma cells rapidly entered apoptosis without IL-6, but mature myeloma cells could survive without IL-6 as well as normal mature plasma cells. Immature myeloma cells as well as myeloma cell lines expressed cyclin D1 mRNA and protein, but not any Cdk inhibitors. On the other hand, mature myeloma cells did not express cyclin D1 but expressed p16, not p21 or p27, as well as normal mature plasma cells. Therefore these results show that immature myeloma cells constitutively express cyclin D1 and can proliferate, and mature myeloma cells as well as normal mature plasma cells preferentially express p16 and can survive for a long time without proliferation. PMID- 9359514 TI - The in vitro effect of pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF) on megakaryopoiesis in normal subjects and patients with myelodysplasia and acute myeloid leukaemia. AB - Mpl ligand is a recently cloned haemopoietic growth factor that stimulates megakaryopoiesis in vitro and in vivo. We describe the in vitro effect of a truncated form of Mpl ligand, recombinant human megakaryocyte growth and development factor (rHuMGDF), on megakaryopoiesis in bone marrow from normal subjects and patients with myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). We used both semi-solid and suspension culture techniques to assess the effect of pegylated (PEG) rHuMGDF on megakaryocyte colony growth (CFU Mk) and on the production of CD61+ cells in 7d suspension cultures. PEG rHuMGDF increased CFU-Mk growth and CD61+ cell production in a dose-dependent fashion in all normal marrows tested. Normal CFU-Mk growth was increased threefold with the addition of 10 ng/ml PEG rHuMGDF to cultures and CD61+ cells were increased 8-10 fold by the same dose. Although increased CFU-Mk growth was only seen in 1/10 AML and 6/16 MDS marrows, CD61+ cell numbers in suspension culture were increased in 9/13 AML and 12/15 MDS samples, responses ranged from very limited to normal magnitude. There was no correlation between platelet count and CFU-Mk number, CD61+ cell number or response to PEG rHuMGDF. We did not find any increased CFU GM colony or cluster growth in response to PEG rHuMGDF and the CD61+ cells produced in suspension culture had features of megakaryocytic differentiation. These data suggest that PEG rHuMGDF can enhance megakaryocyte proliferation in some patients with MDS and AML, and may have a role in the treatment of thrombocytopenia in these patients. PMID- 9359515 TI - Fludarabine ability to down-regulate Bcl-2 gene product in CD5+ leukaemic B cells: in vitro/in vivo correlations. AB - CD5+ B-chronic lymphocytic leukaemia (B-CLL) and mantle cell lymphoma (MCL) in leukaemic phase are characterized by defects in cell death induction that primarily involves the Bcl-2 family of genes. Fludarabine (9-beta-D arabinofuranosyl-2-fluoradenine, F-ara-A) is a potent inducer of apoptosis in CLL cells. This study aimed to determine whether F-ara-A-induced apoptosis might be related to Bcl-2 modifications and to evaluate in vitro/in vivo correlations. Peripheral blood lymphocytes from eight B-CLL and four leukaemic MCL were cultured in the presence of different concentrations of F-ara-A +/- methylprednisolone (MP). F-ara-A down-regulated the expression of Bcl-2 in 5/12 cases. mRNA down-regulation was maximal at 48 h; protein down-regulation was prominent after 48 h. Both events were dose-dependent. The amount of apoptosis was significantly higher in the samples treated with F-ara-A than in those exposed to MP alone. In the seven remaining cases, no Bcl-2 down-regulation was observed after exposure to F-ara-A and the degree of F-ara-A-induced apoptosis overlapped that induced by MP. The in vivo outcome after treatment with three to six courses of F-ara-A was evaluable in 10 patients: 4/5 cases, whose cells had shown in vitro Bcl-2 down-regulation and prominent apoptosis after exposure to F ara-A, had a complete response (CR) and a partial response (PR) was observed in the remaining patient. Of the five patients whose cells had shown no in vitro Bcl 2 modulation after exposure to F-ara-A, two had a PR, but the other three did not show any in vivo clinical response. PMID- 9359516 TI - Fludarabine is effective in the treatment of splenic lymphoma with villous lymphocytes. AB - We report on the use of fludarabine in four patients with splenic lymphoma with villous lymphocytes (SLVL). All four had relapsed after, or failed to respond to, recommended first-line therapies. In each case fludarabine resulted in a complete clinico-haematological response with minimal toxicity, which, in the two patients with long-term follow-up, proved durable. Fludarabine is effective in the treatment of SLVL and should be considered as both a first-line therapeutic option as well as salvage therapy in this condition. PMID- 9359517 TI - Quantification of CMV viraemia in a case of transfusion-related graft-versus-host disease associated with purine analogue treatment. AB - We report a patient who developed transfusion-associated graft-versus-host disease (GvHD) and concurrent cytomegalovirus (CMV) infection, both complications thought to be related to severe T lymphocyte depletion induced by treatment with a purine analogue drug, fludarabine. CMV viraemia was detected by qualitative PCR and the viral load in positive samples was measured using a fully quantitative PCR assay. This quantitative assay enabled the evaluation of the efficacy of antiviral interventions based on the qualitative PCR result. The case illustrates the risks associated with the use of purine analogue drugs, as well as the value of quantitative CMV PCR assays for monitoring CMV infection in immunocompromised patients. PMID- 9359518 TI - Treatment of hairy cell leukaemia (HCL) with 2-chlorodeoxyadenosine (2-CdA): identification of parameters predictive of adverse effects. AB - 2-chlorodeoxyadenosine (2-CdA) induces high complete remission (CR) rates in hairy cell leukaemia (HCL), but is associated with serious toxicities. Therefore we reviewed our experience with 2-CdA in 16 HCL patients, with special attention to adverse effects. One-third of patients presented severe neutropenic infections and/or required prolonged blood support. Patients with low tumour mass and moderate cytopenias were more likely to achieve CR, whereas those with high tumour burden and severe bone marrow impairment were at increased risk of severe infection and blood product requirements. All these unfavourable parameters may be corrected by short-term alpha-interferon (IFN) therapy. Therefore we suggest that patients with unfavourable presenting features might benefit from IFN therapy before 2-CdA. PMID- 9359519 TI - Point mutations of the BCL-6 gene in Burkitt's lymphoma. AB - BCL-6 codes for a transcription factor implicated in chromosomal translocations of diffuse large cell lymphomas. Recent evidence indicates that BCL-6 may also be altered by mutations of its 5' non-coding regions. In this study we have investigated the distribution of BCL-6 5' mutations among 35 Burkitt's lymphoma cases representative of the epidemiologic variants of the disease. Mutations were detected in 6/21 (28.6%) sporadic Burkitt's lymphomas and 7/14 (50%) endemic Burkitt's lymphomas. These data expand the spectrum of B-cell non-Hodgkin's lymphomas associated with BCL-6 5' mutations and have implications for the pathogenesis, histogenesis and clinical monitoring of Burkitt's lymphoma. PMID- 9359520 TI - Adult onset of acute myeloid leukaemia (M6) in patients with Shwachman-Diamond syndrome. AB - Three male patients (two of whom were brothers) with Shwachman-Diamond (SDS) syndrome presented with acute myeloid leukaemia in adulthood. In all three cases there was trilineage myelodysplasia and the morphology was consistent with FAB subtype M6. SDS is an inherited bone marrow failure syndrome with a high propensity to leukaemic transformation. Since this may not occur until adulthood, SDS should be considered in the differential diagnosis of adults presenting with acute myeloid leukaemia, particularly where features of myelodysplasia are prominent. PMID- 9359522 TI - The production of tissue inhibitors of metalloproteinases (TIMPs) in megakaryopoiesis: possible role of platelet- and megakaryocyte-derived TIMPs in bone marrow fibrosis. AB - We quantified tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in serum and plasma in normal control subjects and patients with a low or high platelet count, using one-step sandwich enzyme immunoassays. The serum levels of TIMP-1 and TIMP-2 were 101.1 +/- 13.3 ng/ml, and 82.7 +/- 26.3 ng/ml, respectively, in normal subjects. In patients with an elevated platelet count, such as in essential thrombocytosis, polycythaemia vera, and myelofibrosis, serum levels of TIMP-1 and TIMP-2 were 351.6 +/- 200.9 ng/ml and 148.9 +/- 84.0 ng/ml, respectively. Serum levels of TIMP-1 and TIMP-2 in patients with a low platelet count, such as in aplastic anaemia and idiopathic thrombocytopenic purpura, were 57.2 +/- 25.8 ng/ml and 19.7 +/- 7.68 ng/ml, respectively. The serum level of TIMP-1 was significantly correlated with the platelet count in all subjects. The correlation between the serum level of TIMP-2 and the platelet count was not as strong. The level of TIMP-1 in platelet-depleted plasma was not correlated with the platelet count. Immunohistochemical staining using monoclonal antibodies against TIMP-1 and TIMP-2 showed that megakaryocytes and platelets were positive for both TIMP-1 and TIMP-2, confirming that they are rich sources of TIMPs. TIMP 1 and TIMP-2 stimulated the proliferation of bone marrow fibroblasts, although their effect was less potent than that of TGF-beta and PDGF. Erythroleukaemia and megakaryoblastic cell lines showed the highest secretion of TIMP-1 among the leukaemia cell lines examined. There was no lineage specificity for TIMP-2 secretion. These results suggest that TIMPs released from megakaryocytes or from local platelet coagulation may be important in the development of bone marrow fibrosis. PMID- 9359521 TI - The effects of anagrelide on human megakaryocytopoiesis. AB - Anagrelide, an inhibitor of platelet aggregation, decreases the number of platelets in normal subjects and in patients with myeloproliferative disorders. We describe studies aimed at discovering the general mechanism(s) by which anagrelide acts. We examined three hypotheses: (1) anagrelide shortens platelet survival, (2) anagrelide inhibits the proliferation of megakaryocytic-committed progenitor cells (CFU-M), and (3) anagrelide inhibits maturation of megakaryocytes. We observed that anagrelide did not shorten platelet survival. Proliferation of CFU-M in vivo was not affected by anagrelide, although high concentrations of anagrelide inhibited CFU-M in vitro. In-vivo and in-vitro anagrelide altered the maturation of megakaryocytes, causing a decrease in their size and changing other morphometric features. We conclude that anagrelide decreases the number of platelets primarily by interfering with the maturation of megakaryocytes. PMID- 9359523 TI - Signal transduction involved in the platelet adenylate cyclase sensitization associated with PGH2/TxA2 receptor desensitization. AB - The exposure of human platelets to prostaglandin H2 analogue (PGH2, U46619) induces homologous desensitization and a concomitant adenylate cyclase (AC) sensitization. We demonstrate the involvement of phospholipase C (PLC) in this enzyme sensitization. Pre-incubation of platelets with neomycin, a PLC activity inhibitor, prevented AC sensitization but not PGH2/thromboxane (Tx)A2 receptor desensitization. PGH2/TxA2 receptor desensitization, although necessary, is not sufficient to induce AC sensitization, since neomycin, which prevents AC sensitization, failed to prevent receptor desensitization. Inositol phosphate formation, determined in parallel, was also inhibited. Interestingly, no guanylate cyclase sensitization was noted, suggesting a specific relationship between PGH2/TxA2 receptor desensitization and AC sensitization. In addition, using alkaline phosphatase, a dephosphorylating enzyme, and the tyrosine kinase inhibitor erbstatin, we examined the role of phosphorylation-dephosphorylation on AC sensitization. Effectively, alkaline phosphatase, which has no effect by itself, enhances the cAMP production triggered by prostacyclin in control but not in desensitized platelets. In contrast, erbstatin failed to modify this synthesis, indicating the non-involvement of tyrosine kinase pathway in this process. Our results indicate that the AC sensitization was mediated by PLC and also suggest the participation of other mechanisms, including phosphorylation dephosphorylation processes. This specific enzyme sensitization may be relevant for the in vivo modulation of platelet activation, in different thrombotic diseases with an increased TxA2 generation. PMID- 9359524 TI - Initiation of contact system activation in plasma is dependent on factor XII autoactivation and not on enhanced susceptibility of factor XII for kallikrein cleavage. AB - Various mechanisms have been hypothesized to explain the initiation of contact system activation in plasma. We investigated the capability of dextran sulphate (DS) of different molecular weights to initiate contact system activation in normal human plasma, and compared this with their capability to support factor XII autoactivation and to enhance factor XII susceptibility for cleavage by kallikrein. Dextran sulphate of Mr 500,000 (DS500) and 50,000 (DS50) was able to initiate contact system activation in plasma (determined by measuring the amount of factor XIIa-C1-inhibitor, kallikrein-C1-inhibitor and factor XIa-C1-inhibitor complexes generated) as well as to support factor XII autoactivation and to enhance factor XII susceptibility for cleavage by kallikrein (as measured with amidolytic assays using purified proteins). In contrast, dextran sulphate of Mr 15,000 (DS15) and 5000 (DS5) neither induced contact system activation in plasma, nor supported autoactivation of factor XII, although both of these DS species enhanced the rate of activation of factor XII by kallikrein in the purified system. Based on these properties (i.e. binding of factor XII without inducing autoactivation), DS15 and DS5 were predicted to be inhibitors of contact system activation induced in plasma by DS500, which indeed was observed. We conclude that enhanced factor XII susceptibility for kallikrein activation and factor XII autoactivation are distinct phenomena, the latter being necessary to support activation of the contact system in plasma. PMID- 9359525 TI - Complications experienced with central venous catheters in children with congenital bleeding disorders. AB - The use of central venous catheters (CVC) in children with coagulation disorders allows home treatment, the use of prophylactic blood product replacement and induction of immune tolerance. Previous reports have suggested an almost complete lack of infective complications in this patient group. We reviewed 2 3 patients with bleeding disorders who have had 32 CVC inserted at this institution with a median follow-up of 27 months (range 1-92 months). There were 25 documented line associated infections, including two subcutaneous infections at the port site, and 23 bacteraemias (one episode per 26 patient months at risk). There were 15 Gram-positive, nine Gram-negative and one mixed infection. Infections occurred in 48% of the patients. 15 CVCs were removed: one for erosion through the skin, two for line blockage and 12 for infection. Five patients with inhibitors to factor VIII suffered 14 infections in 12 lines (one per 8.3 months) whereas the 18 without inhibitors suffered 11 infections in 20 lines (one per 50 months) (P<0.03). The use of CVCs is favoured by families of children with bleeding disorders in spite of these complications, but close liaison between families and experienced staff at a Haemophilia Centre is essential to ensure that patients gain the benefits of a CVC as safely as possible. PMID- 9359526 TI - Prevalence of hepatitis G virus RNA in a monocentric population of French haemophiliacs. AB - Hepatitis G virus (HGV) and hepatitis GB virus (GBV-C) have been reported as possible causes of non-A-E transfusional hepatitis. To assess the prevalence of hepatitis G virus infection in haemophiliacs we retrospectively investigated the presence of viral RNA in 92 patients with and without HCV infection. HGV/GBV-C RNA was reverse transcribed and amplified with primers from the 5' non-coding region of the genome. RNA was detected in 16/92 patients (17.4%). Restriction enzyme analysis revealed that the 16 patients belonged to the HGV-like genotype. Serology with E2-specific antibodies demonstrated that HGV viraemia underestimates previous infection by HGV. 33 patients were positive for HGV; all but two have cleared HGV RNA. 47/92 patients had a marker of prior infection by HGV. No difference between HGV RNA positive and negative patients was observed concerning age, diagnosis, HIV and HCV status. Previous HBV infection correlated with the frequency of HGV infection. There was no difference in alanine aminotransferase levels between HGV positive and negative patients. All 18 patients exposed to only virally inactivated plasma-derived concentrates were negative for both HGV RNA and anti E2 antibodies. Prior exposure to untreated concentrates correlated with HGV viraemia (P=0.03), HGV seropositivity (P=0.0002), and markers of HGV infection (P<0.0001). In haemophiliacs with a past exposure to non-inactivated concentrates, persistence of HCV RNA (53/74 patients) was more frequent than HGV RNA persistence (16/74 patients) although HGV viraemia is more frequent than HCV viraemia in blood donors. This may be related to a greater ability of individuals to clear HGV infection and suggests that hepatitis G virus infection in multi-transfused patients has a better outcome than infection with other blood-borne viruses. PMID- 9359527 TI - Spontaneous resolution of p58/EB6 antigen restricted NK-type lymphoproliferative disease of granular lymphocytes: role of Epstein Barr virus infection. AB - We describe a patient with a CD3- lymphoproliferative disease of granular lymphocytes (LDGL) characterized by proliferation of CD3-CD16+ GL, restricted to the expression of p58/EB6 antigen and lacking the p58/GL183 antigen. Using PCR analysis we demonstrated the presence of EBV DNA in the peripheral blood mononuclear cells and purified CD16+ GL from the patient; a monoclonal episomic configuration of the virus could not be demonstrated with Southern blot analysis. The presence of EBV DNA was also detected by PCR in the serum; this finding was associated with a serological pattern consistent with a previous, already seroconverted, EBV infection. During a 4-year follow-up the lymphocytosis spontaneously disappeared; interestingly, in terms of the p58 antigen expression, we provided evidence of the reconstitution of a normal pattern of circulating NK subsets (i.e. p58/EB6+ p58/GL183-, p58/EB6+ p58/GL183+, p58/EB6- p58/GL183-, p58/EB6-p58/GL183+). At the time of resolution of lymphocytosis, EBV-PCR analysis still demonstrated the persistence of EBV DNA in peripheral blood mononuclear cells, but not in the patient's serum. By indicating that inciting agents (in this case EBV) are involved in inducing the GL proliferation, our data contribute insights into the pathogenetic mechanisms accounting for in vivo GL accumulation in LDGL. It appears that a second, still unknown, event is required to determine the neoplastic transformation. PMID- 9359528 TI - Primary familial polycythaemia associated with a novel point mutation in the erythropoietin receptor. AB - Primary familial and congenital polycythaemia (PFCP) is a rare disease characterized by congenital erythrocytosis inherited in an autosomal dominant fashion. Recently, mutations in the erythropoietin receptor (EpoR) have been identified in PFCP families. We describe a Japanese family with an autosomal dominant inheritance of PFCP. An in vitro colony assay demonstrated hypersensitivity of erythroid progenitors to erythropoietin (Epo) in affected family members. Sequence analysis of RT-PCR products amplified from the C terminal region of EpoR transcripts in affected family members revealed that they were all heterozygous for C and T bases at position 5986, which suggested a genetic mutation (C to T) on one allele of EpoR. This mutation gave rise to a translation termination codon TAG at amino acid 435. Thus, the resulting EpoR is a truncated protein product lacking all 74 amino acids downstream of the mutation. To date, all genetic mutations affecting a family with PFCP, including this one, have been located in the cytoplasmic negative regulatory region of the EpoR. All mutations gave rise to truncated Epo receptors between Tyrosine 427 and Tyrosine 455. The phosphotyrosines in this region of EpoR have been demonstrated to be binding sites for SHP-1 phosphatase. Therefore PFCP is presumably brought about as a result of genetic mutations which cause the loss of the SHP-1 binding site in the cytoplasmic region of EpoR. PMID- 9359529 TI - Effect of hepatocyte growth factor on early human haemopoietic cell development. AB - Hepatocyte growth factor (HGF) stimulates cell proliferation, differentiation and migration by binding to its receptor, MET R. Whether the HGF/MET R axis plays an important regulatory role in human haemopoietic cell growth is an unresolved issue. To investigate this situation, we employed several complementary strategies including RT-PCR, FACS analysis, and mRNA perturbation with oligodeoxynucleotides (ODN). We found that very primitive, FACS sorted, CD34+ Kit+ marrow mononuclear cells (MNC) failed to express RT-PCR detectable MET R mRNA. In contrast, MET R expression was easily detectable by RT-PCR in marrow stroma fibroblasts, in cells isolated from BFU-E and CFU-GM colonies, and in unselected normal MNC. Subsequent FACS analysis revealed that MET R protein was detectable on approximately 5% of the latter cells. HGF, at concentrations of 1 50 ng/ml, had no demonstrable effect on survival or cloning efficiency of normal CD34+ MNC in serum-free cultures. Antisense ODN mediated perturbation of MET R mRNA expression in normal CD34+ MNC, with FACS documented decline in protein expression, had no effect on the ability of these cells to give rise to haemopoietic colonies of any lineage. We also examined the biology of HGF/MET R expression in malignant haemopoietic cells. Using the strategies described above, we found that MET R mRNA was expressed in many human haemopoietic cell lines, and that the protein was expressed at high levels on HTLV transformed T lymphocytes. Wild-type CML and AML blast cells also expressed MET mRNA, and HGF was able to co stimulate CFU-GM colony formation in approximately 20% of cases studied. Therefore, although the HGF/MET R axis appears to be dispensable for normal haemopoietic cell growth, it may play a role in the growth of malignant haemopoietic progenitor cells. PMID- 9359530 TI - Automated red cell exchange in sickle cell disease. PMID- 9359531 TI - Characterization of multiple myeloma plasma cells by B-B4 monoclonal antibody. PMID- 9359532 TI - Diagnosis of cobalamin deficiency: the old and the new. PMID- 9359534 TI - TEL/AML1 transcript and p16 gene deletion in a patient with childhood acute lymphoblastic leukaemia. PMID- 9359533 TI - Liver biopsy for haemophilic patients with chronic HCV infection. PMID- 9359535 TI - Frequency of factor V Leiden (Arg506Gln) in France. PMID- 9359536 TI - Effect of bezafibrate treatment over five years on coronary plaques causing 20% to 50% diameter narrowing (The Bezafibrate Coronary Atherosclerosis Intervention Trial [BECAIT]). AB - Recent reports indicate that most coronary events originate from plaques causing <50% diameter stenosis. A subgroup analysis of the Bezafibrate Coronary Atherosclerosis Intervention Trial (BECAIT) data was undertaken to determine the effects of bezafibrate in relation to baseline narrowing. BECAIT included 92 male postacute myocardial infarction patients <45 years of age. Each received double blind treatment with bezafibrate (200 mg 3 times daily) or placebo for 5 years, together with a low-fat diet. Coronary angiography was performed at baseline and after 2 and 5 years. The mean minimum lumen diameter of lesions causing 20% to <50% diameter stenosis at baseline did not narrow over 5 years in the bezafibrate group and decreased by 0.15 mm in the placebo group (p <0.05). In segments with > or =50% diameter stenosis at baseline, no change was seen in either of the 2 groups. In the analysis including only segments with 20% to <50% stenosis at baseline, coronary events were seen in 7 of 40 patients with a progression in minimum lumen diameter of more than the median value and in 3 of 41 patients with a change less than the median value. Thus, bezafibrate had a preferential effect in slowing the progression of narrowings causing <50% stenosis at baseline in young men followed up for a 5-year period after acute myocardial infarction. PMID- 9359537 TI - Rationale, design, and baseline characteristics of a trial comparing aggressive lipid lowering with Atorvastatin Versus Revascularization Treatments (AVERT). AB - This study describes the design, methodologic features, and baseline characteristics of an open-label randomized trial to determine whether aggressive lipid-lowering therapy with atorvastatin is an alternative to angioplasty or other catheter-based revascularization procedures in patients with significant coronary artery disease. Three-hundred forty-one patients with low-density lipoprotein (LDL) cholesterol > or = 115 mg/dl and > or = 1 defined narrowing of a major coronary artery were randomized to atorvastatin or the indicated catheter based revascularization and conventional care (including lipid-lowering therapy if prescribed). Ischemic events are tracked for 18 months. The primary efficacy parameter is the incidence of an ischemic event, defined as 1 of the following: cardiovascular death, cardiac arrest, nonfatal myocardial infarction, the need for coronary bypass grafting or angioplasty, cerebrovascular accident, and worsening angina verified by objective evidence requiring hospitalization (including unstable angina). PMID- 9359538 TI - Admission clinical and electrocardiographic characteristics predicting in hospital development of high-degree atrioventricular block in inferior wall acute myocardial infarction. AB - This study assessed the ability of simple clinical and electrocardiographic variables routinely obtained on admission to identify patients who are at high risk of developing high-degree atrioventricular (AV) block during hospitalization in 1,336 patients with inferior wall acute myocardial infarction (AMI). Patients were classified into 2 initial electrocardiographic patterns based on the J-point to R-wave amplitude ratio: pattern 1: those with J point/R wave <0.5 and pattern 2: patients with J point/R wave > or =0.5 in > or =2 leads of the inferior leads II, III, and aVF. High-degree AV block was found in 6.7% of patients (41 of 615) with pattern 1 versus 11.8% of the patients (85 of 721) with pattern 2 on admission electrocardiogram (p = 0.0008). Multivariate logistic regression analysis revealed that the only variables found to be independently associated with high-degree AV block were female gender (odds ratio [OR] 1.48; 95% confidence interval [CI] 0.98 to 2.23; p = 0.06); Killip class on admission > or =2 (OR 2.24; CI 1.43 to 3.51; p = 0.0004); initial electrocardiographic pattern 2 versus pattern 1 (OR 1.82; CI 1.22 to 2.21; p = 0.003); and absence of abnormal Q waves on admission (OR yes vs no 0.68; CI 0.44 to 1.05; p = 0.08). A simple electrocardiographic sign (J point/R wave > or =0.5 in > or =2 leads) is a reliable predictor of the development of advanced AV block among patients receiving thrombolytic therapy for inferior wall AMI. PMID- 9359539 TI - Significance of prolonged left ventricular wall motion abnormalities after exercise echocardiography following non-Q-wave acute myocardial infarction. AB - Exercise echocardiography was used to assess myocardial ischemia after non-Q-wave acute myocardial infarction in 40 consecutive patients. Resting parasternal long- and short-axis views and apical 4- and 2-chamber views were recorded, digitized, and stored. A maximal symptom-limited exercise test was performed within 21 days (mean 17.7 +/- 3) using a cycle ergometer with continuous monitoring and the echocardiogram was repeated in the same views. Resting and exercise echocardiograms were then compared. Coronary angiography was performed in all patients within 21 days of exercise echocardiography. Stenosis in > or =50% of the lumen diameter was considered significant. Of the 40 patients studied, 29 (72%) had continuing angina and 11 (28%) had no angina. Eighteen patients (62%) with angina developed angina during exercise testing and 19 (65%) developed ST segment depression. In patients without angina, 1 (9%) developed postexercise angina and 2 (18%) developed ST-segment depression. The mean wall motion score index after exercise increased from 1.2 +/- 0.3 to 1.8 +/- 0.4 in patients with continuing angina (p <0.001) and from 1.2 +/- 0.3 to 1.4 +/- 0.3 in patients without angina (p = NS). Prolonged wall motion abnormalities lasting >20 minutes persisted in > or =1 segment in 27 of 29 patients (93%) with angina or in 2 of 1 1 patients (18%) without angina (p <0.001). Patients with continued angina had predominantly 3-vessel coronary artery disease (22 of 29 [76%]) or 2-vessel disease (7 of 29 [24%]), and those without angina had 1-vessel disease (6 of 11 [55%]) or 2-vessel disease (4 of 11 [36%]). One patient had 3-vessel disease. The duration of wall motion abnormality demonstrated a significant relation to 2- and 3-vessel coronary artery disease (p <0.001). Thus, patients with non-Q-wave acute myocardial infarction had a high incidence of multivessel coronary disease not necessarily detected on routine exercise testing. There was also a significant incidence of prolonged wall motion abnormality. PMID- 9359540 TI - Relation of changes in left ventricular peak filling rate during exercise to exercise performance in systemic hypertension and in healed myocardial infarction. AB - Patients with systemic hypertension and coronary artery disease (CAD) often manifest abnormalities at rest in left ventricular (LV) diastolic function and reduced exercise tolerance. It is possible that abnormalities in filling persist during exercise and are partially related to abnormal exercise tolerance. We examined rest and exercise peak filling rate (PFR) to determine if changes in PFR during exercise influence exercise performance. We studied 20 patients with systemic hypertension who had no evidence of CAD (negative thallium-201 stress imaging) and 15 patients with prior myocardial infarction, preserved ejection fraction, and no ischemia by thallium-201 stress imaging. Results were compared with 20 normal subjects. All 55 subjects had rest and exercise radionuclide angiograms Peak workload, exercise time, and LV ejection fraction were reduced in subjects with CAD (57 +/- 24 W, 7.41 +/- 2.91 min, and 60 +/- 9%) compared with subjects with hypertension (72 +/- 21 W, 9.69 +/- 3.03 min, and 70 +/- 6%, p <0.05) and controls (80 +/- 30 W, 10.82 +/- 3.50 min, and 67 +/- 6%, p <0.05). PFR at rest was reduced in CAD subjects (2.40 +/- 0.70 end-diastolic volume per second [EDV/s]) compared with those with hypertension (2.89 +/- 0.70 EDV/s, p <0.02) and controls (3.23 +/- 0.52 EDV/s, p <0.0002). The increments in PFR during exercise were reduced in CAD patients (+1.76 +/- 0.95 EDV/s) compared with hypertensive subjects (+2.93 +/- 1.7 EDV/s) and controls (+3.22 +/- 1.4 EDV/s, p <0.05). The increment in PFR during exercise was related to exercise performance (r = 0.49, p <0.0002). These findings suggest that alterations in LV diastolic filling during exercise are important determinants of exercise performance. PMID- 9359541 TI - Effects of stenting of recent or chronic coronary occlusions on late vessel patency and left ventricular function. AB - Due to high rates of late vessel reocclusion, balloon angioplasty of recent or chronic coronary occlusions is not associated with a sustained improvement in left ventricular function. Recent studies have suggested that stent implantation at coronary occlusions significantly reduces late vessel occlusion. We thus designed a study to analyze the effect of stent implantation at coronary occlusions on late vessel potency and left ventricular function. Twenty-four consecutive patients with recent or chronic coronary occlusions had successful stent implantation and were enrolled in a 6-month angiographic follow-up program. Contrast left ventricular cineangiography, at baseline and 6-month follow-up, as well as preprocedural, postprocedural, and follow-up angiograms analyzed with quantitative angiography were available in 22 of the patients (92%). At follow up, no vessel reocclusion was observed and 32% of the patients, as analyzed by the >50% diameter stenosis criterion, had restenosis. There was a significant improvement in global left ventricular function with a decrease in both left ventricular end-diastolic volume index (LVEDVI, p <0.01) and left ventricular end systolic volume index (LVESVI, p <0.0001) and an increase in left ventricular ejection fraction (LVEF, p <0.0001). Similarly, regional wall motion in the territory of the recanalized artery was also significantly improved (p <0.05). These effects were associated with a reduction in left ventricular filling pressure (p <0.0001). Stent implantation following balloon angioplasty of recent or chronic coronary occlusion is associated with a low rate of late vessel reocclusion, a reduction in cardiac volume, and an increase in ejection fraction. Such effects on left ventricular volumes could have a significant impact on patient survival. PMID- 9359542 TI - Acute and 30-day results of the serpentine balloon expandable stent implantation in simple and complex coronary arterial narrowings. AB - We report the acute and 30-day results with a new serpentine-design, tubular, stainless steel, balloon-expandable stent (beStent) in the first 100 patients. One hundred forty-eight stents were used to treat 103 narrowings in the left anterior descending (n = 46), left circumflex (n = 20), and right coronary (n = 37) arteries. There were 85 de novo and 18 restenotic lesions (lesion length: < 10 mm [31], 10 to 20 mm [43] > 20 mm [29]; lesion type: A [10] B1 [29], B2 [20], C [44]; total occlusions, 23. More than 1 stent was used in 31 patients for treatment of long lesions that could not be covered by 1 stent. The stents used were 15-mm (n = 106), 25-mm (n = 38), or 35-mm (n = 4) long. Stent implantation strategy involved predilatation, deployment, and high-pressure dilatation, using the same balloon if possible. Clinical in-hospital success was 97% (2 patients had stent thrombosis that was recanalyzed, with myocardial infarction developing in 1, and 1 patient died on day 14 from retroperitoneal bleeding treated with surgery and complicated by sepsis). One-month event-free survival was 96%, with 1 death on day 21 due to hypertensive crisis. There were no other major adverse cardiac events in this first complex cohort of patients. In conclusion, the initial experience with this stent demonstrates its safety and efficiency for treating simple and complex coronary disease, with a relatively low rate of complications. Long-term clinical follow-up awaits further investigation. PMID- 9359543 TI - Lead system optimization for transvenous defibrillation. AB - Lead systems that include an active pectoral shell reduce defibrillation thresholds and permit transvenous defibrillation in nearly all patients. A further improvement in defibrillation efficacy is desirable to allow for smaller pulse generators with a reduced maximum output. Accordingly, the purpose of this study was to compare defibrillation thresholds with multiple transvenous lead systems including those with an active pectoral shell to determine which system would optimize defibrillation energy requirements. This prospective study was performed on 21 consecutive patients. Each subject was evaluated with 3 lead configurations with the order of testing randomized. The configurations were a dual coil transvenous lead (lead), the distal right ventricular coil and pectoral pulse generator shell (unipolar), and all 3 components (triad). The right ventricular coil was the cathode for the first phase of the biphasic defibrillation waveform. Delivered energy at defibrillation threshold was 11.2 +/ 3.4 J for the lead configuration, 10.1 +/- 5.2 J for the unipolar configuration, and 7.8 +/- 3.6 J for the triad configuration (p <0.01). Leading edge voltage (p <0.01) and shock impedance (p <0.001) were also decreased for the triad configuration compared with the lead or unipolar configurations, whereas peak current was minimized with the unipolar configuration (p <0.01). We conclude that the combination of a dual coil, transvenous lead and an active pectoral shell reduces defibrillation energy requirements compared with either the lead alone or unipolar configuration. Moreover, the defibrillation thresholds were < or =15 J in all patients using the triad lead system. PMID- 9359544 TI - Electrocardiographic findings in patients with diphenhydramine overdose. AB - QT interval prolongation and torsades de pointes ventricular tachycardia have been reported after therapeutic doses and overdosage of second generation antihistamines, such terfenadine and astemizol. Diphenhydramine (DPHM), a first generation H1 antagonist, is the most frequently used antihistaminic drug. Despite its widespread use, there are no data about cardiac action and electrocardiographic consequences of DPHM overdose. The 12-lead electrocardiograms of 126 patients (mean age 26 +/- 11 years) who had DPHM overdose were evaluated. The ingestion of large doses of DPHM (in majority of cases the dose was >500 mg) was primarily suicidal. Repolarization duration, dispersion, and morphology were evaluated in DPHM overdose patients and compared with those of healthy subjects. Mean heart rate of DPHM overdose patients was 103 +/- 25 beats/min. The QTc duration was significantly longer (453 +/- 43 vs 416 +/ 35 ms, respectively, p <0.001) and mean T-wave amplitude significantly lower (0.20 +/- 0.10 vs 0.33 +/- 0.15 mV, respectively, p <0.001) in DPHM-overdose patients than in control subjects. Dispersion of repolarization was significantly lower in DPHM-overdose patients than in control subjects (42 +/- 25 vs 52 +/- 21 ms, respectively; p = 0.003). None of the DPHM-overdose patients experienced torsades de pointes. In conclusion, DPHM overdose is associated with a significant increase in heart rate and a significant but moderate QTc prolongation. None of the studied patients, including those who had apparent QTc prolongation, experienced torsades de pointes ventricular tachycardia. PMID- 9359545 TI - Incidence of sudden death after radiofrequency ablation of the atrioventricular junction for atrial fibrillation. AB - This study assesses the incidence of sudden death and classifies the causes of death following radiofrequency ablation of the atrioventricular (AV) junction. We studied 220 patients with paroxysmal (n = 105) or chronic (n = 115) atrial fibrillation (AF) and a mean age of 64 +/- 12 years. These patients were followed 31 +/- 15 months after radiofrequency ablation of the AV junction and pacemaker implantation. In 86 patients, structural heart disease was identified before the procedure. All patients were traced via the Swedish National Civic Registry and Cause of Death Registry. The cause-of-death was classified according to data from death certificates, autopsy protocols, and medical records. Thirty-one patients (mean age 69 +/- 11 years, 16 men) died 15 +/- 15 months (range 0.2 to 60) after the procedure. There were 6 sudden unexplained deaths, 14 cardiovascular deaths, and 11 deaths from noncardiovascular causes. Eleven patients, all with structural heart disease, died suddenly out of hospital 16 +/- 16 months (range 0.2 to 42) after the procedure. In 6 of these there was no obvious cause of death. Three of these 6 patients underwent autopsy, which showed extensive coronary artery disease (n = 1), severe heart failure (n = 1) and cardiac hypertrophy and dilation (n = 1). The remaining 3 all had depressed left ventricular systolic function and a history of congestive heart failure. Five of the patients who died suddenly from cardiovascular causes had autopsies that revealed acute myocardial infarction (n = 4) and massive pulmonary embolism (n = 1). PMID- 9359546 TI - Developmental changes of atrioventricular nodal recovery properties. AB - Atrioventricular (AV) nodal recovery properties can be studied by a periodic premature stimulation protocol performed at a slow basic rate. Developmental aspects of these properties have not been determined. The purpose of this study was to determine the developmental changes of AV nodal recovery properties. Forty three children and young adults (male:female ratio 25:18) without AV nodal disease (aged 3.3 to 21.9 years) were studied by delivering premature atrial extrastimuli coupled to basic driven atrial beats. The individual recovery curve was fitted to the equation: A2H2 = A0H0 + exp(alpha -H1A2/tau) for H1A2 > or =theta, where A0H0 is the minimum AH interval, H1A2 is any recovery interval that exceeds the nodal effective refractory period, A2H2 is the corresponding nodal conduction time at any given H1A2, alpha is a constant, tau is the recovery time constant, and theta is the nodal effective refractory period. We found that: (1) A0H0 and alpha constant did not change significantly with age; (2) both tau (r = 0.324; p <0.05) and theta (r = 0.401; p <0.05) had a positive correlation with age; and (3) the maximum change in A2H2 with a 10-ms decrement in H1A2 was 32 ms and did not change significantly with age. Our results suggest that AV nodal recovery properties are age-dependent and both the recovery time constant and effective refractory period lengthen with age. PMID- 9359547 TI - Gender differences in presentation, management, and cardiac event-free survival in patients with syncope. AB - In a MEDLINE search of published English studies (1966 to 1996), no prior study was identified that examined gender-based differences in the management and prognosis of patients admitted with syncope. We studied 109 consecutive patients (48 women) admitted with syncope at the Massachusetts General Hospital (1989 to 1990). All patients underwent Holter monitoring, signal-averaged electrocardiography, and echocardiography according to study protocol. Follow-up was 100% complete (10 +/- 4 months). Women were older (74 +/- 2 vs 66 +/- 2 years, p <0.01) and less likely to have premonitory symptoms when compared with men (46% vs 66%, p <0.05). A greater proportion of men had left ventricular ejection fractions of <0.40 (18% vs 0%, p <0.01), abnormal signal-averaged electrocardiograms (28% vs 8%, p <0.01), and a cardiac cause for syncope (49% vs 25%, p <0.01). Although referral for diagnostic electrophysiologic testing was >3 times as frequent for men compared with women (20% of men vs 6% of women, p <0.05), this difference was not significant after adjustment for age, ventricular arrhythmia, and referral for coronary angiography. During follow-up, 21% of men versus 6% of women (p <0.05) had cardiac events (recurrent syncope, myocardial infarction, or sudden death). Cardiac event-free survival rates were worse for men (p = 0.045). Thus, we have identified gender-based differences in the clinical presentation of syncope for hospital admission. Left ventricular dysfunction and an abnormal signal-averaged electrocardiogram occur more frequently in men. Men are more likely to have cardiac syncope and worse cardiac event-free survival when compared with women. PMID- 9359548 TI - Enteroviral RNA and virus-like particles in the skeletal muscle of patients with idiopathic dilated cardiomyopathy. AB - The role of chronic viral infection in the etiopathogenesis of idiopathic dilated cardiomyopathy (IDC) has generated considerable research. Enteroviruses were the favorite candidates as etiologic agents of IDC. However, enteroviruses were rarely demonstrated in affected hearts. We investigated whether enteroviral infection persists in the heart and in extracardiac sites, particularly in skeletal muscle, in patients with IDC. Blood and myocardial and skeletal muscle samples were collected at cardiac transplantation from 31 IDC patients, 24 non IDC heart disease patients, and 3 heart donors. Samples underwent ultrastructural studies and ribonucleic acid (RNA) extraction. RNA was reverse-transcribed, and 2 nested fragments (bps 179 and 126) were amplified in the highly conserved 5' noncoding region of enteroviral genomic RNA. Enteroviral RNA was found in the skeletal muscle of 12 cases, whereas only 4 hearts (2 of which with positive skeletal muscle) were positive. Of the 24 controls, 2 were positive (1 muscle and heart, 1 muscle only). Automated sequencing confirmed the enteroviral nature of the amplified products. Ultrastructural study showed enterovirus-like particles in 4 of the enterovirus-positive muscles, and myopathic changes in all enterovirus-positive cases. Skeletal muscle hosts chronic enteroviral infection in more than one third of patients with sporadic IDC. Two hypotheses may explain this link. Myocardial damage may derive directly from recurrent subclinical heart infections caused by enteroviruses harbored in skeletal muscle. Alternatively, enterovirus-related myopathy may trigger an autoimmune response to antigens shared by muscle and myocardium. Further studies are needed to assess the importance of these, non-mutually exclusive mechanisms in IDC pathogenesis. PMID- 9359549 TI - Fate of mitral regurgitation following repair of atrioventricular septal defect. AB - The purpose of this study was to evaluate the fate of mitral regurgitation (MR) following repair of atrioventricular septal defects (AVSDs). Echocardiograms of all survivors of isolated AVSD surgery between 1986 and 1996, who had had > or =2 postoperative color Doppler studies (39 patients), were reviewed. On each study, MR severity was graded on a 1+ to 4+ scale, based upon the size of the MR jet. Median age at surgery was 9 months (range 3 to 169); median age at postoperative follow-up was 45 months (range 3 to 107). Mild deterioration of mitral valve function was fairly common. MR severity increased by > or =1 grade in 16 patients (41%) during the course of the study. However, the deterioration in mitral valve function occurred primarily during the early postoperative time intervals. After the initial 32 postoperative months, MR worsened on only 4 occasions and in each instance worsened by only 1 grade. Deterioration to 4+ MR occurred in only 3 patients, and was not observed after the initial 30 postoperative months. Survival curve analysis predicted a 90% probability of not having severe (4+) MR after 30 months (lower 95% confidence bound: 80%). Postoperative MR remains fairly stable following AVSD repair. Serious deterioration is rare, especially after the initial 30 postoperative months. PMID- 9359550 TI - Short- and long-term reproducibility of heart rate variability in patients with long-standing type I diabetes mellitus. AB - Heart rate variability (HRV) has been used to assess cardiac autonomic function noninvasively, understand the pathophysiologic mechanisms of heart disease, evaluate therapy, and assess long-term prognosis. We examined both the short- and long-term reproducibility of the time and frequency domain HRV parameters in 23 type I diabetics over a 12-month interval. Entry criteria included juvenile onset diabetes before age 35 years, >24-year duration of diabetes, diabetes difficult to control, and albuminuria. Standardized noninvasive autonomic testing and 24 hour ambulatory electrocardiographic recordings were obtained. Fifteen men and 8 women (mean age 36.7 years) were enrolled. Fifty-three percent of the men and 75% of the women were smokers, and women had higher cholesterol than men. All HRV parameters were markedly decreased when compared with normal persons. Using Pearson correlation, the time domain indicators of parasympathetic activity demonstrated very strong correlations at 3 and 6 months compared with baseline, with good correlations at 1 year. The average SD of all 5-minute RR intervals maintained a very strong correlation for the entire year (r >0.94). In the frequency domain, the measures of parasympathetic and sympathetic activity maintained a solid correlation for the entire study period. Reproducibility of HRV was also examined using repeated-measures analysis of variance. The time and frequency domain parameters demonstrated very little variation over the study period of 12 months. Thus, our investigation demonstrated that HRV in long-term diabetics using 24-hour ambulatory recordings is abnormal and reproducible over a 12-month interval; very little variation in all HRV parameters, especially in parameters of parasympathetic activity, occurred during the study period. PMID- 9359551 TI - Breath-hold dobutamine magnetic resonance myocardial tagging: normal left ventricular response. AB - Analysis of the changes in myocardial deformation produced by adrenergic stress has been limited by the imaging techniques used. We used rapid magnetic resonance imaging (MRI) myocardial tagging to map the dose-dependent response to incremental dobutamine in the normal human left ventricle. Thirteen volunteers underwent breath-hold tagged cine MRI during dobutamine infusion. Images were acquired throughout systole to a peak dose of 20 microg/kg/min. End-systolic percent circumferential shortening (%S) was measured at 3 transmural locations and 4 circumferential locations at 3 long-axis positions. Mean circumferential shortening velocity (CSV) was also calculated at each location and dose. Mean %S reached a maximum of 26 +/- 3% at 10 microg/kg/min compared with 21 +/- 4% at baseline (p <0.003). Peak %S was reached by 10 microg/kg/min before a significant increase in heart rate or blood pressure and was unchanged at higher doses. In contrast, CSV increased linearly with dobutamine dose from 4.4 +/- 0.9 mm/s at baseline to 9.8 +/- 1.4 mm/s at 20 microg/kg/min (p <0.0001). Breath-hold tagged dobutamine MRI is safe and effective in detecting regional and transmural changes in function during incremental dobutamine. CSV increased continuously across the dobutamine dose range. At low dose (< or =10 microg/kg/min) %S increased without any change in blood pressure or heart rate. Maintenance of peak %S beyond 10 microg/kg/min in the presence of decreasing systolic intervals resulted from a continued increase in CSV. Thus, CSV may be the preferred measure of contractile function during dobutamine stimulation in human myocardium. PMID- 9359552 TI - Beta-blocker prevention of proarrhythmia and proischemia: clues from CAST, CAMIAT, and EMIAT. Cardiac Arrhythmia Suppression Trial. Canadian Amiodarone Myocardial Infarction Arrhythmia Trial. European Myocardial Infarct Amiodarone Trial. PMID- 9359553 TI - Smokers undergoing percutaneous coronary revascularization present with fewer narrowings in the target coronary artery. AB - Among patients undergoing percutaneous coronary revascularization, cigarette smoking remained associated with fewer lesions in the target artery even after adjusting for age, extent of coronary artery disease, diabetes mellitus, and hypertension. These findings support the hypothesis that smokers have less active, yet more active, coronary artery disease. PMID- 9359554 TI - Effects of cardioselective beta blockers on ventilation and gas exchange in patients with heart disease during ramp treadmill testing. AB - The effects of cardioselective beta blockade on ventilation and gas exchange were investigated in 12 male subjects with coronary artery disease during ramp treadmill testing. Patients were able to maintain much of their functional capacity as measured by oxygen consumption in the beta-blocked condition, and also maintained minute ventilation by increasing respiratory rate despite a decrease in tidal volume. PMID- 9359555 TI - Association between plasma homocysteine and coronary artery disease in older persons. AB - The data demonstrate that high plasma homocysteine levels and low plasma folate and vitamin B12 levels are associated with a higher prevalence of coronary artery disease (CAD) in older men and women. Elevated plasma homocysteine levels were observed in 43% of the older men with CAD versus 18% of the older men without CAD, and in 37% of the older women with CAD versus 12% of the older women without CAD. PMID- 9359556 TI - Elevation of soluble adhesion molecules is associated with the severity of myocardial damage in acute myocardial infarction. AB - In 20 patients with acute myocardial infarction, blood samples were taken to study the serial changes in the soluble intercellular adhesion molecule-1 (ICAM 1), vascular cellular adhesion molecule-1 (VCAM-1) and E-selectin. Results indicated that soluble ICAM-1 increased significantly and persisted throughout the study period; however, soluble E-selectin was significantly elevated only transiently and decreased rapidly thereafter, and the VCAM-1 did not increase during entire study period. There was a significant correlation between the levels of ICAM-1, E-selectin 6 hours after admission, and peak creatine kinase level; the levels of ICAM-1 and E-selectin were also positively correlated with total leukocyte count at admission. PMID- 9359557 TI - Polymorphisms of HPA-1 through 6 on platelet membrane glycoprotein receptors are not a genetic risk factor for myocardial infarction in the Japanese population. AB - We examined HPA genotypes derived from polymorphisms of platelet membrane receptors in 88 Japanese patients with early-onset myocardial infarction and in 100 control subjects. The results indicated that HPA genotypes 1 through 6 are not associated with early-onset myocardial infarction. PMID- 9359558 TI - Do diabetes mellitus and systemic hypertension predispose to left ventricular free wall rupture in acute myocardial infarction? AB - Diabetes and systemic hypertension had no influence on left ventricular free wall rupture complicating acute myocardial infarction. Age <65 years and a history of coronary artery disease offers some protection from protection. PMID- 9359559 TI - Effects of a single, daily alcoholic beverage on lipid and hemostatic markers of cardiovascular risk. AB - There is substantial epidemiologic data, but limited experimental data, supporting the mortality benefit of low-dose alcohol consumption. A regimen of a single, daily alcoholic beverage was sufficient to increase both high-density lipoprotein (HDL) (4.4%, p = 0.03) and HDL2 (7.7%, p = 0.04) in men and women, but did not significantly affect hemostatic markers of cardiovascular risk. PMID- 9359560 TI - The effects of reflex parasympathetic stimulation on the QT interval and QT dispersion. AB - The effect of phenylephrine-induced reflex parasympathetic stimulation on QT interval and its dispersion was studied in 16 healthy subjects with a history of paroxysmal supraventricular tachycardia, both during sinus rhythm and during atrial pacing. Results demonstrate that rapid reflex parasympathetic stimulation does not influence QT interval duration or QT dispersion, and also emphasize the inappropriateness of Bazett's formula, the need for comparison of QT intervals during identical heart rates, and the importance of analyzing all 12 leads of a standard electrocardiogram when assessing the effects of various interventions on the QT interval. PMID- 9359561 TI - QT dispersion magnitude is related to the respiratory phase in healthy subjects. AB - Twelve-lead electrocardiograms in 20 healthy volunteers during quiet respiration, maximum inspiration, and maximum expiration were recorded. QT dispersion was statistically significantly shorter both at maximum inspiration and maximum expiration than during quiet breathing. PMID- 9359562 TI - Effect of verapamil on heart rate variability in subjects with normal hearts. AB - Heart rate variability on 24-hour electrocardiographic recording was assessed in 23 patients without structural heart disease before and after 2 months of oral treatment with verapamil prescribed for paroxysmal atrioventricular nodal reentrant tachycardia. Verapamil had no significant effect on overall heart rate variability in the frequency domain, but it increased ultra low frequency power and decreased the low-frequency/high-frequency ratio, deemed to be a marker of sympathetic activity. PMID- 9359563 TI - Usefulness of peak oxygen consumption in predicting outcome of heart failure in women versus men. AB - This investigation finds that percent of predicted maximum oxygen consumption, an age- and gender-adjusted measurement of exercise, capacity, describes the degree of functional impairment in women more accurately than peak oxygen consumption. This evidence must be considered when cardiopulmonary metabolic parameters are used for prognostic stratification of women with heart failure. PMID- 9359564 TI - Application of the continuity equation and valve resistance to the evaluation of St. Jude Medical prosthetic aortic valve dysfunction. AB - Doppler echocardiography was applied to the assessment of patients with surgically documented St. Jude medical aortic valve dysfunction. Derivation of effective orifice area and Doppler velocity index with the continuity equation and calculation of valve resistance accurately differentiated stenotic from regurgitant and normal valves. PMID- 9359565 TI - Features and prognosis of exertional syncope in light-chain associated AL cardiac amyloidosis. AB - Syncope is common in AL amyloid heart disease and in almost 1/3 of our patients who experienced syncope, it was precipitated by physiologic stress. Stress precipitated syncope was associated with a poor prognosis in such patients, both in terms of their median survival of 2 months and was frequently a precursor of sudden cardiac death. PMID- 9359566 TI - Use of a cerebral angiographic catheter facilitates crossing the pulmonary valve in neonates with critical or severe pulmonary valve stenosis. AB - The inability to cross the pulmonary valve in neonates with severe pulmonary stenosis remains a primary reason for procedural failure. We describe the use of a standard pediatric cerebral angiographic catheter that enables crossing the pulmonary valve in these neonates with relative ease and rapidity. PMID- 9359567 TI - Recognition of electrocardiographic left arm/left leg lead reversal. AB - The electrocardiographic error of left arm/left leg lead reversal is difficult to identify. PI amplitude greater than PII as a terminal positive component to PIII may diagnose 90% of such errors. PMID- 9359568 TI - Cardiac lithomyxoma. AB - Among 102 consecutive cardiac myxomas, 10 (10%) showed heavy calcification both on gross inspection and on x-ray examination. Mineralization was partial in 6 patients (mean age 56.8 years) and massive stone-like in 4 (mean age 70 years); relevant histologic findings in patients with partial calcification were Gamma Gandy bodies in 5, cartilaginous and bone metaplasia in 2, and aspecific calcium deposits in 1. All were incidental findings either at autopsy or during clinical investigation. PMID- 9359569 TI - Safety of dobutamine stress echocardiography in patients with left ventricular apical thrombus. AB - We demonstrate that patients with left ventricular mural apical thrombi can safely undergo dobutamine stress echocardiography. These patients also have more severe wall motion abnormalities at rest compared with a group of patients with left ventricular dysfunction without evidence of apical thrombus. PMID- 9359570 TI - Atrial flutter and fibrillation following bee stings. AB - Atrial tachyarrhythmias following bee stings in 2 patients are described. These have not been previously reported in the absence of anaphylaxis. PMID- 9359571 TI - Patch repair of subsemilunar conal septal defect resulting in severe left ventricular outflow tract obstruction. AB - A subsemilunar conal septal defect was closed at 8 months of age with a redundant Dacron patch that bowed into the left ventricular outflow tract, resulting in severe subaortic stenosis and massive left ventricular hypertrophy. Mistaken for cardiomyopathy or myocarditis, this rare complication of subsemilunar ventricular septal defect patch closure led to orthotopic cardiac transplantation followed by death. PMID- 9359573 TI - Characterization of the phosphoglucose isomerase gene from crickets: an analysis of phylogeny, amino acid conservation and nucleotide composition. AB - Although electrophoretic variation in phosphoglucose isomerase (PGI) is often detected in allozyme studies, the Pgi gene has rarely been characterized. Here, I present the cDNA sequence of the Pgi gene from two species of field cricket, Gryllus pennsylvanicus (U65475) and G. veletis (U65476), in which the PGI protein is suspected of being under balancing selection. Phylogenetic analyses support the conclusion that these sequences are truly cricket Pgi. The cricket amino acid sequences are compared to sequences from other taxa to determine conserved residues that may be essential for the function of the protein. Such analysis is necessary as there is no well-resolved structure of the PGI protein. In addition, the compositional bias of cricket Pgi is different from the other animal Pgi genes characterized to date. PMID- 9359572 TI - The best hospitals in the USA for heart disease--1997. PMID- 9359574 TI - Esterase gene amplification in Culex pipiens. AB - In the mosquito Culex pipiens one of the major resistance mechanisms to organophosphorous pesticides (OPs) is increased detoxification of insecticide. This resistance is the consequence of overproduction of two types of esterases, esterases A and B, coded at two loci, Est-3 (A esterase) and Est-2 (B esterase). We have analysed the genomic structure of these genes in different strains resistant to OPs and have attempted to characterize the different types of mutations leading to the resistant phenotypes. It is shown that, concerning the more frequent resistant phenotypes, mutations leading to resistance are of two main types. First, overproduction of one A esterase present in Southern France results from a regulatory mechanism. The second type of mutation is gene amplification which involves events that have initially generated the duplication of both the A and B esterase or only the B esterase locus. We report the point that the most frequent esterase overproductions are the results of eight different mutations and that, given the range of distribution of these genotypes, mutation leading to an efficient resistance gene is one of the most limiting factors for the evolution toward resistance in Culex pipiens. PMID- 9359575 TI - Cloning and characterization of three Musca domestica yolk protein genes. AB - The yolk protein (yp) genes encode the major nutritional polypeptides deposited in developing oocytes for subsequent utilization during embryogenesis, and represent a highly conserved family of genes in higher Diptera. Originally isolated from Drosophila melanogaster, they are expressed in a temporal-, tissue- and sex-specific manner in all species in which they have been identified. We report here the isolation of cDNAs encoding three independent yolk proteins from the common housefly, Musca domestica. Expression of the three M. domestica yp genes is analysed both by Northern and in-situ hybridization. We discuss in an evolutionary context both the significance of the expression patterns, and regions of apparent polypeptide sequence divergence. PMID- 9359576 TI - The white gene of the tephritid fruit fly Bactrocera tryoni is characterized by a long untranslated 5' leader and a 12kb first intron. AB - A 300 bp fragment from exon 6 of the white gene of Bactrocera tryoni was used to screen a B. tryoni genomic library. One positive (approximately 14 kb) insert contained exons 2-6 of white by nucleotide and amino acid sequence similarity to the white genes of D. melanogaster (O'Hare et al., 1984; Pepling & Mount, 1990). Lucilia cuprina (Garcia et al., 1996). Ceratitis capitata (Zwiebel et al., 1995) and Anopheles gambiae (Besansky et al., 1995). A white 5' cDNA fragment containing exons 1, 2 and part of exon 3 was amplified, cloned and sequenced. An inverse PCR fragment of genomic DNA was generated, containing the exon 1 coding region plus approximately 2.1 kb of upstream sequence, encompassing the putative promoter of the gene. Exon 1 was found to be 728 bp long, encoding the first twenty-five amino acids. The full length of intron 1 was shown to be 12 kb (amplified using long PCR protocols), up to 3 times the length of the longest white intron 1 isolated to date. PMID- 9359577 TI - Mosquito clathrin heavy chain: analysis of protein structure and developmental expression in the ovary during vitellogenesis. AB - We have deduced the amino acid sequences of clathrin heavy chain (CHC) polypeptides based on cDNA and genomic clones from the mosquito, Aedes aegypti. Two isoforms which differ in the very beginning of the N-terminal domain, ovary specific AaCHCa and somatic-specific AaCHCb, were identified, characterized and compared to one another as well as to CHC polypeptides from different species. The 1682 amino acid sequence of the AaCHCa isoform predicts a molecular mass (M[r]) of 191,743 daltons and an isoelectric point of 5.80, whereas the 1674 amino acid sequence of the AaCHCb isoform predicts a M(r) of 191,033 daltons and an isoelectric point of 5.71. Both mosquito AaCHC isoforms are highly conserved, with full-sequence identities of 88% to Drosophila melanogaster, 81% to mammal (rat, cow and human), 71% to C. elegans, 58% to Dictyostelium discoideum, and 49% to yeast CHC polypeptides. The highest degree of conservation is in the middle portion of the mosquito CHC molecule which includes the linker region and extended triskelion arm, with decreasing conservation through the N-terminal domain, trimerization domain, and the relatively diverged C-terminal region. The protein domains do not directly correspond to specific exons of the mosquito AaCHC gene, with the exception of exon 6 which encodes the C-terminal domain of the CHC polypeptide. Polyclonal antibodies raised against a bacteria-expressed AaCHC fusion protein recognized one major band of about 180 kDa in vitellogenic ovary whole-lysate. Immunogold labelling of the AaCHC polypeptide localized it to the coat of coated pits and coated vesicles in oocytes from vitellogenic follicles. Northern blot and in situ hybridization analyses suggest that regulation of AaCHC gene expression in the ovary is complex, and it likely involves both developmental and hormonal signals. PMID- 9359578 TI - Double-strand conformation polymorphism (DSCP) analysis of the mitochondrial control region generates highly variable markers for population studies in a social insect. AB - Genetic markers were obtained for the termite Nasutitermes corniger by DSCP (double-strand conformation polymorphism) analysis of PCR-amplified mitochondrial control region DNA. This procedure revealed twenty-one haplotypes in forty-four colonies, whereas a restriction fragment length polymorphism analysis detected only nine haplotypes. Sequence analysis of DSCP fragments of contrasting mobilities suggests that the electrophoretic haplotypes are caused by DNA curvature in this highly AT-rich region. DSCP markers showed that some termite colonies contained maternally unrelated queens, each of which produced worker offspring. This pattern is consistent with nest founding by unrelated queens. Due to the availability of conserved primers for the mtDNA control region, DSCP analysis may readily reveal comparatively high levels of variation in a wide variety of organisms. PMID- 9359579 TI - Molecular identification of sympatric chromosomal forms of Anopheles gambiae and further evidence of their reproductive isolation. AB - Three chromosomal forms of Anopheles gambiae s.s., designated as Bamako, Mopti and Savanna, were studied for diagnostic PCR assays based on the analysis of the X-linked ribosomal DNA (rDNA). The study was performed on a 1.3 kb fragment containing part of the 28S coding region and part of the intergenic spacer region. The amplified material was cut with fourteen restriction enzymes to detect Restriction Fragment Length Polymorphisms (RFLPs). The enzymes Tru9I and HhaI produced patterns of DNA bands which differentiated Mopti from Savanna and Bamako; moreover, a distinct 'hybrid' pattern was recognized in the F1 female progeny from the cross of Mopti with either one of the other two chromosomal forms. The diagnostic significance of the PCR-RFLP assay was verified on 203 karyotyped females from field samples collected in two villages in Mali and one village in Burkina Faso. Agreement was observed between the chromosomal and the molecular identifications. No 'hybrid' molecular patterns were detected even among carriers of rare heterokaryotypes hypothetically produced by crosses between Mopti and Savanna. The results confirm previous observations indicating barriers to gene flow within An. gambiae s.s. and supporting the specific status of the taxonomic units proposed on cytogenetic ground. PMID- 9359580 TI - Cloning and characterization of a serine protease from the human malaria vector, Anopheles gambiae. AB - The nucleotide and deduced amino acid sequence of a serine protease (AgSp24D) from the human malaria vector, Anopheles gambiae, is presented. The gene product is a 271 amino acid protein that contains the conserved serine, histidine and aspartic acid residues found in serine proteases, and has the highest identity to a serine protease of unknown function from Drosophila melanogaster. In situ hybridization to the polytene chromosomes detects a single band at 24D. Northern analysis reveals only low levels of transcripts in larvae and pupae, but more abundant transcription products occur in adults. Interestingly, this analysis also shows that adult males express much higher levels of AgSp24D mRNA than females. In addition, Plasmodium-refractory mosquitoes express higher levels of AgSp24D mRNA than susceptible mosquitoes although the biological significance of this remains to be examined. The thorax is the primary site for expression in the adults. The lack of a dramatic increase in AgSp24D mRNA levels following blood feeding suggests that this protease is not involved in digestive processes. Transcriptional induction does not follow cold shock, septic wounding, bacterial injection, laminarin injection or CM-Sephadex bead injection. PMID- 9359581 TI - Intracerebral microdialysis study of glutamate reuptake in awake, behaving rats. AB - The central nervous system has high-affinity uptake systems for the clearance of amino acid transmitters. These systems are found in both neurons and astrocytes. Previous studies have shown that the uptake of amino acid transmitters by astrocytes in culture can be modulated by adrenergic agents. The objectives of this study were to develop a methodology that evaluates the brain's reuptake capacity for glutamate in awake, behaving animals and to determine whether glutamate reuptake is under alpha-adrenergic regulation in the intact central nervous system. Male Sprague-Dawley rats weighing 250-450 g were used in this study. The extraction fraction of L-[3H]glutamate with [14C]mannitol as a reference was measured. The cortical extraction fraction of L-[3H]glutamate corrected for [14C]mannitol (EL-glu) reaches steady state rapidly and is both stable and repeatable. EL-glu is a measure of L-glutamate reuptake and not metabolism. EL-glu is decreased in a dose-dependent manner by the addition of the glutamate reuptake blocker D,L-threo-beta-hydroxyaspartic acid or unlabeled L- glutamate. In addition, EL-glu is increased in a dose-dependent manner by the alpha1-adrenergic agonist phenylephrine, and this increase is blocked by the alpha-adrenergic antagonist phentolamine. PMID- 9359583 TI - Gender difference in hypothalamic-pituitary-adrenal axis response to alcohol in the rat: activational role of gonadal steroids. AB - Alcohol administration activates the hypothalamic-pituitary-adrenal (HPA) axis of both male and female rats, with females secreting more adrenocorticotropin (ACTH) and corticosterone than males in response to the same dose of alcohol. Our earlier work suggested that this gender difference arises due to the activational effects of gonadal steroids. In particular, we hypothesized that both androgens and estrogens play a role, with androgens exerting an inhibitory influence while estrogens elevate activity of the HPA. In the present studies, we tested this hypothesis by manipulating steroidal milieu in male rats using surgical castration and chronic implantation of testosterone (T), dihydrotestosterone (DHT), or estradiol (E2). Intact male and female rats were included as controls. Injection of alcohol (3 g/kg b.wt., i.p.) resulted in elevation of blood alcohol levels, ACTH and corticosterone in all groups. However, the amount of ACTH secreted was greater in females and castrated males implanted with E2 than in intact males. In castrated males, regardless of androgen implantation, the ACTH response was intermediate, with mean levels between those of females and males, but not differing significantly from either. In contrast to the ACTH results, significantly higher corticosterone secretion was measured in females and castrated males which did not receive a steroid implant. Since there were no significant differences between groups in blood alcohol levels (BALs), these results are not due to steroid-dependent alterations in alcohol metabolism. Because the ACTH data confirmed an activational effect of E2, we sought to determine whether this steroid regulated levels of corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) mRNAs in the paraventricular nucleus of the hypothalamus (PVN). Four pretreatment groups were studied: intact males, intact females, castrated males, and castrated males implanted with E2. Two weeks after surgery, alcohol or vehicle was administered 3 h before brains were collected. In intact males, alcohol treatment elevated levels of both CRF and AVP mRNAs in the PVN, as previously reported. In contrast, this treatment decreased CRF mRNA in castrated males implanted with E2. In addition, steroid pretreatment alone elevated CRF mRNA levels in castrated males. Although we did not observe E2 dependent increases in CRF or AVP mRNAs, our data do support a complex effect of gonadal steroids on expression of these mRNAs in the PVN. PMID- 9359584 TI - CNS sites involved in sympathetic and parasympathetic control of the pancreas: a viral tracing study. AB - The viral transneuronal tracing method was used to identify the CNS cell groups that regulate the parasympathetic and sympathetic outflow systems of the pancreas. Pseudorabies virus (PRV) was injected into the pancreas of vagotomized rats and after 6 days survival, the pattern of transneuronal labeling in the CNS sympathetic regulatory regions was determined. The converse experiment was performed in order to elucidate the central parasympathetic cell groups that regulate the pancreas. Immunohistochemical methods were used to identify putative neuropeptide- and catecholamine-containing CNS neurons involved in these regulatory circuits. The major finding of this study indicates that five brain regions, viz., paraventricular hypothalamic nucleus, perifornical hypothalamic region, A5 catecholamine cell group, rostral ventrolateral medulla, and lateral paragigantocellular reticular nucleus, contain a considerable amount of overlap in cell body labeling. In addition, the ventrolateral part of the periaqueductal gray matter and gigantocellular reticular nucleus, ventral part also showed a similar overlap, but the numbers of neurons found in these areas were considerably lower than the five major regions. These data suggest that these brain regions may provide parallel and possibly redundant, autonomic pathways affecting glucagon and adrenaline release. PMID- 9359585 TI - Oscillatory fast wave activity in the rat pyriform cortex: relations to olfaction and behavior. AB - Bursts of rhythmical fast waves (> 1 mV, peak frequency approximately 16 Hz; mean frequency approximately 20 Hz) are elicited in the olfactory bulb and pyriform cortex in waking or urethane-anesthetized rats (1.25 g/kg, i.p.) by olfactory stimulation with organic solvents (xylene, toluene, methyl methacrylate, oil of turpentine) or components of anal gland secretions of rat predators (2 propylthietane, weasel; trimethyl thiazoline, red fox). These waves are specifically related to olfaction since they: (a) are blocked when the nares are sealed; (b) are not elicited by non-olfactory stimuli; (c) are unrelated to concurrent motor activity; and (d) can only be elicited in anesthetized tracheotomized rats when an odorous airstream is drawn through the nasal passages. Pyriform fast waves appear to be somewhat specific to the odors of organic solvents and predators as other strong odors (ammonia, caproic and butyric acids, cadaverine) are ineffective. During natural sleep or after treatment with scopolamine hydrobromide, low voltage pyriform background activity is replaced by larger amplitude, irregular 1-20 Hz waves. The scopolamine-induced waves are not blocked by spontaneous motor activity. We suggest that the pyriform cortex, like the archicortex and the neocortex, receives a cholinergic activating input. PMID- 9359582 TI - Opioid and non-opioid NMDA-mediated predator-induced analgesia in mice and the effects of parasitic infection. AB - The present study examined the nociceptive responses (50 degrees C, hot-plate) of uninfected and subclinically parasitized male mice exposed to the odor of a predator, an ecologically relevant threatening stimulus. In uninfected mice a 15 min exposure to 2-propylthietane, the major component of weasel odor, induced a naloxone-reversible opioid analgesia. A 30-s exposure elicited a shorter duration and lower amplitude 'non-opioid' analgesia that was insensitive to naloxone, partially sensitive to either the serotonin-1A (5-HT1A) agonist, 8-OH-DPAT, or the GABAA antagonist, bicuculline, and blocked by the competitive N-methyl-D aspartate (NMDA) antagonist, NPC 12626. In contrast, mice chronically (25 days) and subclinically infected with the murine nematode, Heligmosomoides polygyrus, failed to show a significant non-opioid analgesia and displayed a markedly lower level of opioid analgesia than uninfected mice. These results suggest that NMDA receptor mechanisms are potently associated with the expression of the analgesia arising from exposure to the naturally aversive stimulus of predator odor. These findings also demonstrate that parasites, and likely other subchronic infections, can have a significant impact on the display of opioid and non-opioid stress induced analgesia arising from exposure to the ethologically relevant stimulus of predator odor. PMID- 9359586 TI - Increased expression of extracellular signal regulated kinase 1 after axotomy in the dorsal motor nucleus of the vagus nerve and the hypoglossal nucleus. AB - The extracellular signal regulated kinases (Erks) cascade is a major signalling system by which cells transduce extracellular signals into intracellular responses. To obtain information about the role of Erks in retrograde neuronal reaction, we investigated the changes of Erk 1 and Erk 2 with in situ hybridization and immunohistochemical study in the dorsal motor nucleus of vagus nerve, which shows degenerative changes, and the hypoglossal nucleus, which shows regenerative changes, of adult rats after axotomy. The expression of mRNA and protein of Erk 1 increased between 7 and 28 days after axotomy both in the vagal and hypoglossal nuclei, however, there was no remarkable change in those of Erk 2. The increased expression of Erk 1 is common to both regenerative hypoglossal and degenerative vagal neurons. These findings indicate that Erk 1 is closely related with the retrograde neuronal reaction but whether neurons are destined to survive or die depends on some other factors. PMID- 9359587 TI - Effects of volatile anaesthetics on the membrane potential and ion channels of cultured neocortical astrocytes. AB - Volatile anaesthetics cause changes in the membrane resting potential of central neurons. This effect probably arises from actions on neuronal ion channels, but may also involve alterations in the ion composition of the extracellular space. Since glial cells play a key role in regulating the extracellular ion composition in the brains of mammals, we analyzed the effects of halothane, isoflurane and enflurane on the membrane conductances and ion channels of cultured cortical astrocytes. Astrocytes were dissociated from the neocortex of 0-2-day old rats and grown in culture for 3-4 weeks. Anaesthetic-induced changes in the membrane potential were recorded in the whole cell current-clamp configuration of the patch-clamp technique. We further studied the effects of halothane and enflurane on single ion channels in excised membrane patches. At concentrations corresponding to 1-2 MAC (1 MAC induces general anaesthesia in 50% of the patients and rats), membrane potentials recorded in the presence of enflurane, isoflurane and halothane did not differ significantly from the control values. At higher concentrations, effects of enflurane and halothane, but not of isoflurane, were statistically significant. Single-channel recordings revealed that halothane and enflurane activated a high conductance anion channel, which possibly mediated the effects observed during whole cell recordings. In less than 10% of the membrane patches, volatile anaesthetics either increased or decreased the mean open time of K+-selective ion channels without altering single-channel conductances. In summary, it seems unlikely that the actions of volatile anaesthetics described here are involved in the state of general anaesthesia. Statistically significant effects occurred at concentrations ten times higher than those required to cause half-maximal depression of action potential firing of neocortical neurons in cultured brain slices. However, it cannot be excluded that the changes observed in the membrane conductance of cortical astrocytes disturb the physiological function of these cells, thereby influencing the membrane resting potential of neurons. PMID- 9359588 TI - Mu and delta opioids but not kappa opioid inhibit voltage-activated Ba2+ currents in neuronal F-11 cell. AB - Whole-cell patch-clamp recordings were used to study Ba2+ currents through voltage-dependent Ca2+ channels in dorsal root ganglion x mouse neuroblastoma hybrid (F-11) cells. Opioid agonists selective for either mu (Tyr-D-Ala-Gly-Mephe Gly-ol; DAMGO) or delta (Tyr-D-Pen-Gly-Phe-D-Pen-OH; DPDPE) receptors inhibited high-threshold Ba2+ currents. The inhibition was reversible, naloxone-sensitive, and dose-dependent. The inhibitory effects of both DAMGO and DPDPE were blocked by pretreatment of the cells with pertussis toxin (PTX) as well as by brief exposure to the sulfhydryl alkylating agent, N-ethylmaleimide (NEM). The N-type Ca2+ channel antagonist omega-conotoxin GVIA (omega-CTX GVIA) irreversibly inhibited high threshold Ba2+ currents by 66% and blocked the inhibitory effect of DAMGO or DPDPE. In contrast, the L-type Ca2+ channel blocker nifedipine inhibited high threshold Ba2+ currents by 15% and failed to block the inhibitory effect of DAMGO or DPDPE. These results demonstrate that mu and delta opioid receptors are negatively coupled to N-type Ca2+ channels via PTX- and NEM sensitive GTP-binding proteins in F-11 cells. PMID- 9359589 TI - Immunohistochemical studies on glutamatergic, GABAergic and glycinergic axon varicosities presynaptic to parasympathetic preganglionic neurons in the superior salivatory nucleus of the rat. AB - After the superior salivatory nucleus (SSN) neurons were labeled by administration of cholera toxin B subunit (CTB) or wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) to the pterygopalatine ganglion, morphological interactions between SSN neurons and fibers afferent to SSN neurons were examined by light and electron microscopy with double-immunostaining techniques. Antibodies to either the neurotransmitters or its receptor were used to label glutamatergic, GABAergic and glycinergic synapses on these neurons. By light microscopy, SSN neurons were identified in the ipsilateral ventrolateral part of the rostral medulla oblongata, and rich distributions of glutamate- and GABA-immunoreactive (ir) axon varicosities were observed around SSN neurons. Electron microscopy revealed that dendrites of SSN neurons received asymmetric synapses from glutamate-ir axon varicosities. Somata as well as dendrites received symmetric synapses from GABA-ir varicosities, or showed immunoreactivity for glycine receptors. Quantitative analysis by electron microscopy showed that glutamate-ir axon varicosities comprised 45.3% of total axon profiles in the SSN region, while GABA-ir varicosities were 20.8% and varicosities presynaptic to glycine receptors were 19.9%. These findings suggest that glutamatergic, GABAergic and glycinergic inputs, originated from a variety of nuclei, directly affect the activity of SSN neurons, and play a role in the regulation of the pterygopalatine ganglion of the rat. PMID- 9359590 TI - A rat model of focal embolic cerebral ischemia. AB - We developed a new model of embolic cerebral ischemia in the rat which provides a reproducible and predictable infarct volume within the territory supplied by the middle cerebral artery (MCA). The MCA was occluded by an embolus in Wistar rats (n = 71). An additional three non-embolized rats were used as a control. Cerebral blood flow (CBF) was measured by means of laser Doppler flowmetry (LDF) and perfusion weighted imaging (PWI) before and after embolization. The evolution of the lesion was monitored by diffusion weighted imaging (DWI). Cerebral vascular perfusion patterns were examined using laser scanning confocal microscopy. Infarct volumes were measured on hematoxylin and eosin (H&E) stained coronal sections. The lodgment of the clot at the origin of the MCA and the ischemic cell damage were examined using light microscopy. Regional CBF in the ipsilateral parietal cortex decreased to 43 +/- 4.1% (P < 0.05) of preischemic levels (n = 10). Confocal microscopic examination revealed a reduction of cerebral plasma perfusion in the ipsilateral MCA territory (n = 6). MRI measurements showed a reduction in CBF and a hyperintensity DWI encompassing the territory supplied by the MCA (n = 4). An embolus was found in all rats at 24 h after embolization. The infarct volume as a percentage of the contralateral hemisphere was 32.5 +/- 3.31% at 24 h (n = 20), 33.0 +/- 3.6% at 48 h (n = 13), and 34.5 +/- 4.74% at 168 h (n = 12) after embolization. This model of embolic focal cerebral ischemia results in ischemic cell damage and provides a reproducible and predictable infarct volume. This model is relevant to thromboembolic stroke in humans and may be useful in documenting the safety and efficacy of fibrinolytic intervention and in investigating therapies complementary to antithrombotic therapy. PMID- 9359591 TI - Formalin-evoked Fos expression in spinal cord is enhanced in morphine-tolerant rats. AB - It has been hypothesized that tolerance to the analgesic effects of morphine results from the development of a compensatory response in neurons that express the opioid receptor or in neural circuits in which those neurons participate. The compensatory response establishes a sensitized state in these neurons. To determine if administration of a noxious stimulus can unmask a sensitization of dorsal horn neurons in morphine-pelleted rats, we injected morphine-tolerant and control rats with formalin into the plantar surface of the hindpaw, counted the number of flinches for 2 h and then processed the lumbar cord for Fos immunocytochemistry. Although there was no significant difference in flinching behavior between the morphine-tolerant and control groups, we recorded significantly increased total Fos-like immunoreactivity at the L4/5 and L2 segments both ipsilateral and contralateral to the site of formalin injection in the morphine-tolerant rats compared to the control rats. These results suggest that lumbar spinal cord neurons are sensitized during the development of tolerance, that the sensitization can be unmasked by the administration of a noxious stimulus and that it is manifested as increased expression of the Fos protein in the lumbar cord. PMID- 9359592 TI - The GABA(B) receptor antagonist CGP36742 attenuates the baclofen- and scopolamine induced deficit in Morris water maze task in rats. AB - Effects of CGP36742 (3-aminopropyl-n-butylphosphinic acid), an orally active GABA(B) receptor antagonist, on the baclofen- and scopolamine-induced deficit of place learning in the Morris water maze task were examined in rats. Rats were given four training trials per day with the submerged platform at a fixed location in the maze for 4 days. On day 4, the rats were required to swim in the pool without the platform after the fourth training trial (probe test). Intraperitoneal injection of baclofen (4 mg/kg) or scopolamine (0.3 mg/kg) significantly increased the escape latency to reach the platform and decreased the duration in the quadrant where the platform had been originally located. Increased latency in the training trials and decreased duration in the probe test induced by baclofen or scopolamine were significantly attenuated by oral administration of CGP36742 at doses of 10 and 30 mg/kg. In the rotarod test, CGP36742 at a dose of 100 mg/kg but not at doses of 10 or 30 mg/kg antagonized the baclofen-induced motor incoordination. Thus, there was dissociation between the effective doses of CGP36742 in the learning task and those in the sensory motor test. These results suggest the possible involvement of cholinergic systems as well as GABA(B) receptor systems in the CGP36742 action. PMID- 9359593 TI - A chronic MPTP model reproducing the slow evolution of Parkinson's disease: evolution of motor symptoms in the monkey. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been shown to induce parkinsonism both in man and non-human primates. Several models have now been developed, but acute MPTP administration does not consistently reproduce all the clinical features of the disease. To mirror the slow evolution observed in human pathology, a chronic model of intoxication is necessary. The present study describes a chronic MPTP protocol in the monkey. Six monkeys received daily injections of MPTP (0.2 mg/kg i.v.) until they reached a score over 8 on the clinical rating scale (15.5 days +/- 1.1). Full parkinsonism was first obtained on the 22nd day. Levodopa testing (20 mg/kg per os) alleviated motor abnormalities (51%), proving the parkinsonian nature of these disturbances. Histological lesions reproduced those observed in Parkinson's disease with a decrease in tyrosine hydroxylase immunoreactivity of 90%. This model so could be of great interest for the study of the dynamic physiopathological changes which occur in Parkinson's disease and consequently for research on new neuroprotective therapies. PMID- 9359594 TI - Recovery of striatal dopamine function after acute amphetamine- and methamphetamine-induced neurotoxicity in the vervet monkey. AB - In six vervet monkeys, presynaptic striatal dopamine function was assessed longitudinally by [18F]fluoro-L-DOPA (FDOPA)-positron emission tomography (PET) after administration (2 x 2 mg/kg, i.m., 4 h apart) of either amphetamine (Amp), n = 3, or methamphetamine (MeAmp), n = 3. At 1-2 weeks postdrug, both Amp and MeAmp exposure effected similar decreases (60-70%) in the FDOPA influx rate constant (FDOPA Ki), an index of striatal dopamine synthesis capacity. Subsequent studies in these subjects showed that FDOPA Ki values were decreased by 45-67% at 3-6 weeks, by 25% at 10-12 weeks and by 16% in one Amp-treated subject at 32 weeks. Biochemical analysis showed that striatal dopamine concentrations were decreased by 75% at 3-4 weeks and by 55% at 10-12 weeks. These results indicate that in vervet monkey striatum, an acute Amp or MeAmp drug dosage produces extensive striatal dopamine system neurotoxicity. However, these effects were reversible; observed time-dependent recovery in both FDOPA Ki and dopamine concentrations indicates that neurochemical plasticity remains active in the adult primate striatum. At 3-4 and 10-12 weeks postdrug, the concurrent characterization of the striatal FDOPA Ki and dopamine concentrations for individual subjects showed that Ki decreases between 24 and 67% corresponded to dopamine depletions of 55-85%. These relatively larger postdrug decrements in steady-state striatal dopamine concentrations suggest that compensatory increases in dopamine synthesis capacity develop in the partially lesioned striatum. In contrast to the dopamine depletion in striatum, substantia nigra concentrations remained unchanged from referent values at both 3-4 and 10-12 weeks postdrug. Thus, the integrity of the substantia nigra could not be inferred from decreases in the striatal FDOPA Ki parameter. This disparity between striatum and substantia nigra reactivity to systemic administration of amphetamines suggests that each has unique dopamine system regulatory mechanisms. PMID- 9359595 TI - Effects of prenatal exposure to ethanol on callosal projection neurons in rat somatosensory cortex. AB - The distribution and density of callosal projection neurons in the somatosensory cortex of mature rats was altered by prenatal exposure to ethanol. The density of callosal neurons was significantly greater in ethanol-treated rats than in controls. Ethanol exposure also altered the laminar distribution of callosal projection neurons. Whereas in control rats the cell bodies of callosal projection neurons were in layers II/III and V, in ethanol-treated rats most of these neurons were distributed in layers V and VI. Many of the ectopic neurons were generated toward the end of cortical neuronogenesis (i.e., on gestational day 20). This contrasts with controls wherein co-generated cohorts were distributed in layer II/III. Thus, the connectional phenotype of the callosal projection neurons is retained regardless of its laminar residence. These ethanol induced abnormalities apparently result from defects in neuronal migration and axonal pruning. PMID- 9359596 TI - Overexpression of alpha and beta isoforms of Ca2+/calmodulin-dependent protein kinase II in neuroblastoma cells -- H-7 promotes neurite outgrowth. AB - Since the alpha and beta isoforms of CaM kinase II are known to be expressed almost exclusively in the brain, we compared the effect of overexpression of the beta isoform of CaM kinase II with that of the alpha isoform. The subcellular distribution of the alpha isoform was different from that of the beta isoform, although the catalytic properties of the alpha and beta isoforms expressed in transfected cells were similar to those of brain CaM kinase II. The alpha isoform was found in the soluble fraction more than in the particulate fraction, whereas most of the beta isoform bound to subcellular structures. In the cell overexpressing alpha and beta isoforms of CaM kinase II, neurite extension was promoted when compared with the morphology of neo transfectants. Neurite outgrowth of cells overexpressing CaM kinase II was further stimulated by the treatment of 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), a selective but not absolutely specific inhibitor of protein kinase C. The morphological change was rapid and observed within 1 h followed by H-7 treatment. Morphological changes, such as the number of cells with neurites and length of neurites were greater in the beta cells than in the alpha cells. Chelerythrine, a specific inhibitor of protein kinase C, also stimulated the neurite outgrowth of these cells. Some substrates of CaM kinase II related to neurite outgrowth were detected in cells overexpressing CaM kinase II stimulated with H-7. These results suggest that CaM kinase H and protein kinase C play an important role in the control of cell change, and that the subcellular distribution of CaM kinase II is important for regulating cellular functions efficiently. PMID- 9359597 TI - Long-term consequences of neonatal exposure to diazepam on cerebral glucose utilization, learning, memory and anxiety. AB - The long-term consequences of neonatal exposure to diazepam (DZP) on behavioral abilities and local cerebral glucose utilization (LCGU) in 12 brain regions involved in the control of memory and anxiety were studied in adult rats. Rat pups received a daily subcutaneous injection of 10 mg/kg DZP or of the dissolution vehicle from postnatal day (P) 2 to 21. Learning and memory were tested in P60-P70 rats over 5 consecutive days in a T maze and an eight-arm maze while anxiety and reaction to novelty were tested in a two-compartment box with a two-step staircase on the enriched side. LCGU was measured in the P60 rat by the quantitative autoradiographic [14C]deoxyglucose method. In the T maze, when performed without delay between the two trials, the rate of alternation was significantly lower in DZP- than in vehicle-exposed rats on the first 2 days of testing and similar in both groups on days 3-5. In the procedure with a 30 s intertrial delay, the rate of alternation was similar in DZP- and vehicle-treated rats on all days of testing. In the eight-arm maze, DZP-treated rats were more active, i.e., entered more arms per minute than control animals. The number of arms entered before the first error was lower on day 1 and higher on day 3 in DZP compared to vehicle-exposed rats. In the two-compartment box, DZP-treated rats crossed more often and spent more time than controls on the lower step of the staircase while control rats made more rearings and spent more time than DZP exposed rats in the well protected corner of the box. LCGU were decreased by early DZP exposure in six regions which were mammillary body, septum, visual and prefrontal cortices, dorsomedian caudate nucleus and mediodorsal thalamus. In conclusion, postnatal DZP treatment induced at adulthood an increase in activity, a delay in task acquisition but no learning-memory impairment and reduced the level of anxiety allowing active responding to novelty. These quite subtle behavioral changes were accompanied by discrete metabolic decreases in regions mediating anxiety, reflecting a change in the level of anxiety and emotionality. PMID- 9359598 TI - Nicotinic acetylcholine sensitivity of rat petrosal sensory neurons in dissociated cell culture. AB - Using whole-cell, patch-clamp techniques we investigated acetylcholine (ACh) sensitivity of dissociated sensory neurons from rat petrosal ganglia after 4 h-14 days in vitro. In approx. 68% of petrosal neurons (PN; n = 109) ACh, applied by fast perfusion or pressure ejection from a 'puffer' pipette, caused a rapid depolarization associated with a conductance increase. Under voltage clamp near the resting potential (approx. - 60 mV), ACh induced a hexamethonium-sensitive, inward current (IACh), mimicked by nicotine application, suggesting the presence of neuronal nicotinic acetylcholine receptors (nAChR). The reversal potential of IACh occurred near 0 mV (n = 4), a region where the I-V curve displayed a prominent rectification. The dose-response relation for IACh versus ACh concentration was fitted by the Hill equation with EC50 = approx. 33.9 microM and Hill coefficient = approx. 1.6. The activation phase of IACh was well fitted by a single exponential with mean (+/- S.E.M.) time constant of 102 +/- 82 ms (n = 6); the desensitization phase of IACh was best fitted by the sum of two exponentials, with time constant of 870 +/- 210 ms (n = 6) and 8576 +/- 1435 ms (at -70 mV). Fluctuation analysis yielded an apparent single-channel conductance of 21.6 +/- 10 pS (mean +/- S.E.M.; n = 4). These data indicate that a major subpopulation of sensory neurons in visceral petrosal ganglia of the rat express nAChR. Thus, if similar receptors are present on corresponding nerve terminals, they could mediate fast afferent excitation in response to ACh released at peripheral targets, e.g., the chemosensory carotid body. PMID- 9359600 TI - Long-term enhancement of entorhinal-dentate evoked potentials following 'modified' ECS in the rat. AB - Electroconvulsive therapy (ECT) is widely used as a treatment for drug-resistant depression. The animal analogue of ECT is electroconvulsive shock (ECS) seizures. We have recently shown that repeated ECS seizures cause a long-lasting, perhaps permanent, enhancement in entorhinal-dentate evoked potentials in the rat. Our study, however, involved 'unmodified' ECS, whereas in clinical practice ECT is now usually given in its 'modified' form (with near-threshold currents, a short acting barbiturate, muscle relaxant and oxygen). We have therefore repeated our experiments using modified ECS. Entorhinal-dentate evoked potentials were measured in Long-Evans rats before and after: (1) eight modified ECS seizures; or (2) eight sham modified ECS trials. Despite the use of the modified procedure, a significant and long-lasting enhancement in population spike amplitude was seen in the ECS group. We conclude that the modified procedure does not protect rats against the long-lasting enhancement of evoked potentials. Similar changes may be occurring in the brains of patients subjected to modified ECT. PMID- 9359599 TI - Excitation of rat subthalamic nucleus neurones in vitro by activation of a group I metabotropic glutamate receptor. AB - The subthalamic nucleus (SThN) provides a glutamate mediated excitatory drive to several other component nuclei of the basal ganglia, thereby significantly influencing locomotion and control of voluntary movement. We have characterised functionally the metabotropic glutamate (mGlu) receptors in the SThN using extracellular single unit recording from rat midbrain slices. SThN neurones fired action potentials spontaneously at a rate of 10 Hz which was increased by the group I/II mGlu receptor agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate (1S,3 R-ACPD; 1-30 microM) and the group I selective agonist (S, R) dihydroxyphenylglycine (DHPG; 1-30 microM). However, both the group II selective agonist (1S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV; 1 microM) and the group III selective agonist (S)-2-amino-4-phosphonobutanoic acid (L-AP4; 10 microM) were without effect, indicating that the excitation was mediated by a group I mGlu receptor. The excitation caused by DHPG (3 microM) was reversed by co-application of the mGlu receptor antagonist (+)-alpha-methyl-4 carboxyphenylglycine (MCPG; 500 microM). Thus a group I mGlu receptor mediates excitation of SThN neurones, and suggests a use for group I mGlu receptor ligands for treatment of both hypo- and hyperkinetic disorders of basal ganglia origin, such as Parkinson's disease and Huntington's disease. PMID- 9359603 TI - Folate and 10-formyltetrahydrofolate dehydrogenase (FDH) expression in the central nervous system of the mature rat. AB - 10-Formyltetrahydrofolate dehydrogenase (10-FTHFDH) is a folate-binding protein that is important in folate metabolism. In addition, 10-FTHFDH catalyzes the rate limiting step in hepatic folate-dependent formate oxidation. We measured folate concentrations and examined cellular 10-FTHFDH expression in regions of the adult rat central nervous system (CNS), to study components of CNS oxidative formate metabolism. Folate was detected in every CNS region studied; the concentrations ranged from 3 to 14% of that detected in the liver. Immunohistochemical expression of 10-FTHFDH was identified in many CNS structures. 10-FTHFDH was mostly expressed by glia, especially astrocytes and ventricular ependyma. Neuronal expression was weak but evident in the cerebral cortex, basal ganglia, cerebellum, and spinal cord. Thus, CNS tissue has the chief components of folate dependent formate oxidation and the chief site of this oxidation appears to be glia. PMID- 9359602 TI - A 0.1-700 Hz current through a voltage-clamped pore: candidate protein for initiator of neural oscillations. AB - A protein of mass 7643 Da and sequence identical to that of subunit c, the pore part, of the mitochondrial adenosine triphosphate synthase complex, was co purified with cholesterol in crystals formed from a chloroform/methanol extract of bovine brain gray matter plasma membranes. Reconstitution of the protein containing crystals in phospholipid bilayers and assay of current by patch-clamp analysis, showed an oscillating cation current at constant voltage, typically of frequency 0.5-200 Hz. The ceroid-lipofuscinoses state in mammals and man (Batten disease), in which subunit c accumulates in lysosomes, affords a rich source of the protein. Pure subunit c from affected sheep liver (in the absence of cholesterol) was also assayed, the current displaying identical sodium oscillations to those of brain crystals. The results suggest that if a protein similar to subunit c resides in the plasma membrane of neural cells, it could be responsible for spontaneous oscillations in brain tissue. The relevance of these results to the pathogenesis of Batten disease is discussed. PMID- 9359601 TI - Subcellular localization of the K+ channel subunit Kv3.1b in selected rat CNS neurons. AB - Voltage-gated potassium channels constitute the largest group of heteromeric ion channels discovered to date. Over 20 genes have been isolated, encoding different channel subunit proteins which form functional tetrameric K+ channels. We have analyzed the subcellular localization of subunit Kv3.1b, a member of the Kv3 (Shaw-like) subfamily, in rat brain at the light and electron microscopic level, using immunocytochemical detection. Detailed localization was carried out in specific neurons of the neocortex, hippocampus and cerebellum. The identity of Kv3.1b-positive neurons was established using double labeling with markers for specific neuronal populations. In the neocortex, the Kv3.1b subunit was expressed in most parvalbumin-containing bipolar, basket or chandelier cells, and in some bipolar or double bouquet neurons containing calbindin. In the hippocampus, Kv3.1b was expressed in many parvalbumin-containing basket cells, as well as in calbindin-positive neurons in the stratum oriens, and in a small number of interneurons that did not stain for either parvalbumin or calbindin. Kv3.1b protein was not present in pyramidal cells in the neocortex and the hippocampus, but these cells were outlined by labeled presynaptic terminals from interneuron axons that surround the postsynaptic cell. In the cerebellar cortex, granule cells were the only population expressing the channel protein. Careful examination of individual granule cells revealed a non-uniform distribution of Kv3.1 staining on the somata: circular bands of labeling were present in the vicinity of the axon hillock. In cortical and hippocampal interneurons, as well as in cerebellar granule cells, the Kv3.1b subunit was present in somatic and unmyelinated axonal membranes and adjacent cytoplasm, as well as in the most proximal portion of dendritic processes, but not throughout most of the dendrite. Labeling was also seen in the terminals of labeled axons, but not at a higher concentration than in other parts of the axon. The distribution in the cells analyzed supports a role in action potential transmission by regulating action potential duration. PMID- 9359604 TI - Histamine modulates NMDA-dependent swelling in the neostriatum. AB - In the present study, infrared differential interference contrast videomicroscopy was used to examine the effect of histamine on N-methyl-D-aspartate-induced swelling in neostriatal neurons in a brain slice preparation. Histamine caused a concentration-dependent increase in swelling evoked by N-methyl-D-aspartate. By itself, histamine did not cause swelling. Electrical stimulation also caused N methyl-D-aspartate-dependent swelling which was enhanced by histamine. In addition, histamine was found to enhance N-methyl-D aspartate-induced swelling from postnatal day 7 to 28 but not at postnatal day 3. Finally, this histamine induced enhancement was prevented by treatment with either the H2 receptor antagonist cimetidine or with the potassium channel blocker tetraethylammonium chloride. Overall, these findings suggest that histamine modulates N-methyl-D aspartate receptor function in the neostriatum through a H2 receptor-mediated regulation of potassium channels. PMID- 9359605 TI - Focal cerebral ischemia enhances glial expression of ecto-5'-nucleotidase. AB - The effect of ischemia on the reactive expression of ecto-5'-nucleotidase in rat brain was studied 6 h and 1, 2 and 7 days after permanent middle cerebral artery occlusion (MCAO). The distribution of 5'-nucleotidase in the infarcted brain was compared to markers for astrocytes (glial fibrillary acidic protein (GFAP)) and microglia (complement receptor type 3, antibody OX42) using histological staining or immunohistochemistry. 5'-Nucleotidase could be associated with reactive astrocytes by immunohistochemistry and with reactive microglia by enzyme histochemistry. In the untreated control 5'-nucleotidase was associated with astrocytes only in the hippocampus and the submeningeal space. After ischemia the enzyme was expressed on reactive astrocytes in the tissue surrounding the volume of infarction. Individual reactive astrocytes were observed 6 h after MCAO and the astrocytic expression became continuously enhanced during the following days. An enzyme histochemical analysis of 5'-nucleotidase activity revealed a postischemic increase in reaction product around the infarcted tissue. Seven days after MCAO a discrete band (0.2-0.4 mm) of reaction product characterized the rim of the infarcted area. This band of activity of 5'-nucleotidase colocalized with a band of immunoreactivity for OX42, indicative of an intense accumulation of 5' nucleotidase expressing microglia. Our results suggest that ischemia following permanent MCAO results in an upregulation of the capacity for the hydrolysis of nucleotides within the tissue adjacent to the infarcted volume. Nucleotides released from the damaged cells can be hydrolyzed and the adenosine eventually formed may exert neuroprotective functions limiting the extent of damage. PMID- 9359606 TI - Changes in the H-reflexes of ankle extensor and flexor muscles at the initiation of a stepping movement in humans. AB - The underlying neural mechanisms of the silencing of electromyographic (EMG) activity observed between antagonistic muscles of the human ankle joint preceding a fast stepping movement were examined. Twelve subjects were asked to perform a unilateral stepping movement under a reaction time condition while standing upright. A silent phase (SP) which was composed of the time difference between the offset of tonic soleus (Sol) activity and the onset of phasic tibialis anterior (TA) activity was observed consistently in all subjects. At the time interval corresponding to the SP, inhibition of the Sol Hoffmann reflex (H reflex) and facilitation of the TA H-reflex took place. In nine of the 12 subjects, disynaptic reciprocal Ia inhibition (Ia inhibition) from TA Ia fibers to the Sol motoneuron (MN) pool increased before the TA EMG onset. We also observed a clear distinction between the changes in the time course of the Ia inhibition and that of long latency reciprocal (D1) inhibition in six of the 12 subjects. However, when the subjects performed the same motor task with their backs supported against a wall wherein the tonic Sol activation disappeared, no changes in the H-reflex occurred. These results suggest that these changes, occurring at the segmental level, might be controlled by the supraspinal motor center in a feed-forward manner during anticipatory postural adjustment. The functional significance would be to counterbalance the disturbance from an intentional forthcoming movement in order to maintain an adequate posture. PMID- 9359607 TI - Modulation of H reflexes in human tibialis anterior muscle with passive movement. AB - Significant movement-induced gain changes in H reflexes have been observed in soleus muscle following passive movement of the lower limb. Hypotheses from these concepts were tested on magnitudes of H reflexes in tonically contracted tibialis anterior. From eleven subjects at rates of 20 and 60 r.p.m. passive leg movement, statistically significant attenuation from controls and phasic modulation occurred. The results make more general the conclusions from soleus H reflexes. However, the functional effect should be much smaller, as tibialis anterior H reflexes are smaller compared to those in soleus. PMID- 9359608 TI - Subdiaphragmatic vagotomy does not prevent fever following intracerebroventricular prostaglandin E2: further evidence for the importance of vagal afferents in immune-to-brain communication. AB - Brain-mediated sickness responses can be blocked by subdiaphragmatic vagotomy, suggesting that vagal afferents signal peripheral inflammation or infection. This study tested whether subdiaphragmatic vagotomy disrupts sickness responses by interrupting effector pathways. If this explanation is correct, intracerebroventricular prostaglandin E2-induced fever should be blocked by this procedure. Fever was unaffected by subdiaphragmatic vagotomy, thus these data provide support for the conclusion that vagal afferents signal the brain during immune activation. PMID- 9359609 TI - Hippocampal cholinergic blockade enhances hypothalamic-pituitary-adrenal responses to stress. AB - We examined the role of the hippocampal cholinergic system, which is known to mediate processes related to fear and anxiety, in the regulation of stress induced hypothalamic-pituitary-adrenal (HPA) activity. Bilateral intra hippocampal injections (30 microg per side) of the muscarinic antagonist Scopolamine augmented adrenocorticotropin and corticosterone responses to restraint without altering basal HPA activity compared to vehicle-treated animals. These results suggest that the hippocampal cholinergic system regulates stress-induced HPA activity and may serve to coordinate behavioral and neuroendocrine responses to stress. PMID- 9359610 TI - Organization of the lamprey striatum - transmitters and projections. AB - The purpose of the present study is to characterize the striatum of the lamprey by immunohistochemical and tracing techniques. Cells immunoreactive for GABA and substance P (SP), and positive for acetylcholinesterase, are present in the lamprey striatum. Immunoreactive (ir) fibers were detected by antisera raised against SP, dopamine, enkephalin and serotonin. These immunoreactive fibers were mainly located in the periventricular neuropil that borders the striatum and in which GABAergic striatal neurons distributed their dendritic arbors. Putative connections between the striatum, the ventral part of the lateral pallium, and the diencephalic motor centers involved in the control of locomotion were studied by using fluorescein-coupled dextran amines (FDA) as a tracer. The striatum projects to the ventral part of the lateral pallium (lpv), where GABA-ir cells and SP-ir fibers were also present. The lpv in turn projects to the ventral thalamus, which has descending connections to the reticulospinal cells involved in the control of locomotion. These results, together with previous findings of histaminergic and neurotensin projections, suggest that the lamprey striatum and its inputs with regard to neurotransmitters/modulators are very similar to those of modem amniotes, including primates, and are thus conserved to a high degree. PMID- 9359611 TI - Hydrogen peroxide opposes the hypoxic depression of evoked synaptic transmission in rat hippocampal slices. AB - Hydrogen peroxide (H2O2, 3.3 mM) partially reversed the hypoxic depression of the evoked population spike recorded from CA1 region of rat hippocampal slices. It is known that elevated endogenous adenosine contributes to the hypoxic inhibition of the population spike. Exogenous adenosine (100 microM) inhibited the population spike that had been partially resuscitated by H2O2 during maintained hypoxia. It is concluded that the ability of H2O2 to oppose hypoxic depression does not occur at the level of the adenosine receptor since added adenosine was still effective in inhibiting the evoked potential in the presence of H2O2. PMID- 9359612 TI - The effect of a peripheral mononeuropathy on immunoreactive (ir)-galanin release in the spinal cord of the rat. AB - The pattern of ir-galanin release in the spinal cord of rats with a peripheral mononeuropathy was studied. On the side of the cord ipsilateral to the nerve injury enhanced ir-galanin release was found in the superficial dorsal horn. It is probable that, after nerve injury, some primary afferent neurons spontaneously release galanin from their central terminals. PMID- 9359613 TI - Audiogenic seizures after neck tourniquet-induced cerebral ischemia in the rat. AB - Development of audiogenic seizures (AGS) and their correlation with neurodegeneration were studied after 7.5 min of whole-brain ischemia. One day post-ischemia, all animals became hyperreactive and responded to auditory stimulation by generalized seizures. Neuronal necrosis developed already 6 h post ischemia in inferior colliculi, reticular thalamic nucleus and hippocampal hilar region. Repeated ischemia did not induce any neurological changes, suggesting that the neurological effects are consequences of selective neuronal injury. PMID- 9359614 TI - Neocortical and hippocampal electrical activities are similar in spontaneous and cholinergic-induced REM sleep. AB - Neocortical and hippocampal EEG power spectra obtained during REM-like sleep induced by unilateral carbachol microinjections (0.01 M, 0.02 M and 0.2 M; volume 20 nl) into the ventral part of the nucleus reticularis pontis oralis have been compared with EEG power spectra obtained during spontaneous REM sleep. Our findings indicate that neocortical and hippocampal electrical activities during the REM-like state generated by carbachol delivery in this pontine region mimic those present in spontaneous REM sleep. PMID- 9359615 TI - GABA injected into the anterior dorsal tegmentum (ADT) of the midbrain blocks stepping initiated by stimulation of the hypothalamus. AB - Previous work showed that the activity rates of certain neurons in the anterior dorsal tegmentum (ADT) of the midbrain correlated with the onset of stepping elicited by hypothalamic stimulation. This study determined if reversible inactivation of the ADT would block locomotion elicited by hypothalamic stimulation of anesthetized rats (urethane, 800 mg/kg). GABA (concentrations 0.25 1.0 mg/microl in saline) were injected in 52 sites in 21 rats. GABA at volumes of 0.1 or 0.2 microl blocked hindlimb stepping in 18 cases. Locomotor blocks occurred within 5 min of the injection, and typically recovered within 10-20 min. The effective blocking sites were clustered around the interstitial nucleus of the medial longitudinal fasciculus. Sites more dorsal and more anterior were not as effective as sites in and ventral to this nucleus. The data are consistent with a role for the ADT of the midbrain in locomotor initiation. PMID- 9359616 TI - Brain neurosteroids during the mouse oestrous cycle. AB - Concentrations of the neuroactive steroid 3alpha,5alpha-tetrahydroprogesterone (TH PROG or allopregnanolone) and its precursors progesterone (PROG) and 5alpha dihydroprogesterone (DH PROG) have been measured in mouse brain throughout the oestrous cycle. Plasma PROG concentrations were also measured for comparison. At each stage, circadian fluctuations were found in the concentrations of brain PROG and its metabolites. Such fluctuations were greater than those attributable to any particular stage of the oestrous cycle. Over the entire cycle, a significant correlation was found between brain TH PROG (or DH PROG) and PROG concentrations but not between brain TH PROG (or DH PROG) and plasma PROG concentrations. There was also no correlation between endogenous TH PROG (or DH PROG) and activity of the 5alpha-reductase converting 3H-PROG to 3H-DH PROG in whole brain homogenates. Concentrations of another neuroactive steroid, pregnenolone sulphate (PREG S), in the brain during the oestrous cycle were in phase with plasma PROG but not brain PROG concentrations. Our results indicate that circadian and ovarian influences on the concentrations of PROG and its metabolite TH PROG in female whole mouse brain are caused predominantly by changes in the supply of PROG from within the tissue, whatever the contribution of peripheral sources. PMID- 9359617 TI - Structural requirement for axonal regeneration-promoting effect of polyamines in cultured rat hippocampal neurons. AB - We have previously found that spermine, spermidine and putrescine promote axonal regeneration following axotomy in cultured rat hippocampal neurons. In the present study, we investigated which part of the polyamine molecule is responsible for the regeneration-promoting effect. Testing the effects of several synthetic analogues revealed that the butanediamine moiety is essential for the activity and the terminal primary amines are necessary for full agonist activity. The structure-activity relationship indicates that the regeneration-promoting effects of polyamines are not associated with NMDA receptors. PMID- 9359618 TI - Swelling and damage of glial cells by lactacidosis and glutamate: effect of alpha trinositol. AB - The therapeutical efficacy of alpha-trinositol (D-myo-inositol-1,2,6 trisphosphate), an isomer of the intracellular messenger IP3, was analyzed on cytotoxic swelling and damage of glial cells in vitro from lactacidosis or glutamate. C6 glioma cells suspended in a physiological medium were either exposed to pH 5.0 by administration of lactic acid, or to 1 mM glutamate. Cell swelling and viability were quantified by flow cytometry. Lactacidosis of pH 5.0 led to an increase in cell volume to 139.7 +/- 1.3% within 20 min whereas alpha trinositol was reducing the swelling response by approximately 25% (P < 0.01). In addition, at pH 5.0 the fraction of viable cells was lowered from 94.3 +/- 0.2% (control) to only 53.8 +/- 3.1% after 60 min. Alpha-trinositol was found to protect also cell viability; at 60 min of lactacidosis 70.2 +/- 1.6% of the cells still were viable (P < 0.01). The addition of glutamate (1 mM) to the cell suspension led to a steady increase in cell size, reaching 110% of control at 120 min, irrespectively of whether alpha-trinositol was added or not. PMID- 9359619 TI - Ultrastructural study of the GABAergic and cerebellar input to the nucleus reticularis tegmenti pontis. AB - The nucleus reticularis tegmenti pontis is an intermediate of the cerebrocerebellar pathway and serves as a relay centre for sensorimotor and visual information. The central nuclei of the cerebellum provide a dense projection to the nucleus reticularis tegmenti pontis, but it is not known to what extent this projection is excitatory or inhibitory, and whether the terminals of this projection contact the neurons in the nucleus reticularis tegmenti pontis that give rise to the mossy fibre collaterals innervating the cerebellar nuclei. In the present study the nucleus reticularis tegmenti pontis of the cat was investigated at the ultrastructural level following anterograde and retrograde transport of wheat germ agglutinin coupled to horseradish peroxidase (WGA-HRP) from the cerebellar nuclei combined with postembedding GABA immunocytochemistry. The neuropil of this nucleus was found to contain many WGA HRP labeled terminals, cell bodies and dendrites, but none of these pre- or postsynaptic structures was double labeled with GABA. The vast majority of the WGA-HRP labeled terminals contained clear spherical vesicles, showed asymmetric synapses, and contacted intermediate or distal dendrites. Many of the postsynaptic elements of the cerebellar afferents in the nucleus reticularis tegmenti pontis were retrogradely labeled with WGA-HRP, while relatively few were GABAergic. We conclude that all cerebellar terminals in the nucleus reticularis tegmenti pontis of the cat are nonGABAergic and excitatory, and that they contact predominantly neurons that project back to the cerebellum. Thus, the reciprocal circuit between the cerebellar nuclei and the nucleus reticularis tegmenti pontis appears to be well designed to function as an excitatory reverberating loop. PMID- 9359620 TI - Modeling drug-melanin interaction with theoretical linear solvation energy relationships. AB - The affinity of drugs and other xenobiotic agents for melanin is a well-known phenomenon, often occurring with serious physiological consequences. For example, the interaction of anti-psychotic drugs with neuromelanin may play a pivotal role in the induction of extrapyramidal movement disorders associated with the chronic administration of phenothiazine and other neuroleptic agents. Little, however, is known about the complete nature of melanin-drug binding and the impact of these interactions on the physico-chemical properties of melanin. Data, such as binding affinities, can be analyzed using recently developed computational methods that combine mathematical models of chemical structure with statistical analysis. In particular, theoretical linear solvation energy relationships provide a convenient model for understanding and predicting biological, chemical, and physical properties. By using this modeling technique, drug-melanin binding of a set of 16 compounds has been analyzed with correlation analysis and a set of theoretical molecular parameters in order to better understand and characterize drug-melanin interactions. The resulting correlation equation supports a charge transfer model for drug-melanin complex formation and can also be used to estimate binding constants for related compounds. PMID- 9359621 TI - X-ray microanalysis and phagocytotic activity of cultured retinal pigment epithelial cells in hypoxia. AB - PURPOSE: To investigate the influence of the functional and morphological changes induced in retinal pigment epithelial (RPE) cells by retinal ischemia, we evaluated the phagocytotic activity, the concentration of various elements, and ultrastructure in cultured RPE cells in hypoxia. METHODS: The concentrations of oxygen in incubators were adjusted to 20, 10, and 1% by the addition of nitrogen for 72 hr. To observe phagocytotic activity and its relationship to actin filaments, the filaments of RPE cells incubated with fluoresbrite carboxylate YG microspheres were stained with rhodamine phalloidin. Some of the specimens were subjected to X-ray microanalysis by scanning electron microscope after being fixed, freeze-dried, and coated with carbon to investigate the cytoplasmic concentration of elements. A part of the latter specimens was also observed by transmission electron microscope after being embedded in epon and cut into ultrathin sections to see the ultrastructural changes inside cell. RESULTS: Lowering oxygen concentrations from 20% to 1% swelled RPE cells and decreased the number of fluoresbrite carboxylate YG microspheres phagocytized by RPE cells. Phagocytosis of a large amount of latex beads (30 microl) for 24 hr in 1% oxygen caused a disruption of RPE cells. Na, S, and P were detected in RPE cells cultured in 20% oxygen. Reducing the oxygen concentration from 20 to 10 or 1% significantly decreased Na and increased S. Mitochondria were observed in RPE cells in 20 and 10% oxygen, but many vacuoles were observed in the cytoplasm in 1% oxygen. CONCLUSION: Hypoxia as low as 1% oxygen induced malfunction of phagocytosis and the fragility of RPE cells. We could speculate the imbalance of the electrolytes such as Na or a decrease of antioxidants such as glutathione containing S as a reason of disturbance of cell viability. PMID- 9359622 TI - Genetic and biochemical studies on ommochrome genesis in an albino strain of a terrestrial isopod, Armadillidium vulgare. AB - Genetic studies and quantitative determination of levels of 3-hydroxykynurenine and kynurenine were performed in an albino strain of a terrestrial isopod Armadillidium vulgare. From the results of matings between the albino and the albino, the red, the dark red, or the wild type individuals, the albino A. vulgare seems to be regulated by an autosomal gene(s) recessive to its wild allele. Litter mating of F1 progenies obtained by crossing the albino and the red mutant or the albino and the dark red mutant yielded progenies at a ratio of 3:6:3:4 for the red, the dark red, the wild, and the albino phenotypes, respectively. The albino gene(s) seems not to be allelic but to be epistatic to the red gene(s) with respect to ommochrome biosynthesis. Quantitative determination of 3-hydroxykynurenine carried out by high-performance liquid chromatography with electrochemical detection revealed that the 3 hydroxykynurenine content in the albino was significantly lower than that in the wild or the red type. The whole content of 3-hydroxykynurenine after enzymatic conversion of kynurenine to 3-hydroxykynurenine was still considerably lower than that found in the wild type, even though it increased after the conversion. The albino gene(s) seems to be associated with a blockage at distinct level(s) of ommochrome biosynthesis. PMID- 9359623 TI - The amphibian Kupffer cells build and demolish melanosomes: an ultrastructural point of view. AB - This ultrastructural research was carried out to investigate the nature of the liver pigment cells of anuran and caudate amphibians, the pattern of melanosome ontogenesis, and the demolition processes of old melanosomes. We demonstrate that these liver pigment cells are able to internalize zymosan particles and latex beads; therefore, being professional phagocytes, they, as liver resident macrophages, can be classified as Kupffer cells (KCs). They show "melanosomogenesis centers" in which several maturation stages of premelanosomes are visible; the premelanosomes are formed by two principal components: a filamentous structure that will constitute the "inner" area of the melanosome and a vesicular component, budding from the Trans Golgi Network and that carries enzymes, which will constitute the "cortical area" of the melanosome. Thus the KCs, thanks to the presence of the "melanosomogenesis centers," are also melanosome producing cells. They are also able to demolish melanosomes by heterophagocytosis and, probably, also by autophagocytosis. In conclusion, we propose a classification of vertebrate pigment cells. PMID- 9359624 TI - Characterisation of ACTH peptides in human skin and their activation of the melanocortin-1 receptor. AB - Alpha-Melanocyte-stimulating hormone (alpha-MSH) is a proopiomelanocortin (POMC) derived peptide, which is produced in the pituitary and at other sites including the skin. It has numerous effects and in the skin has a pigmentary action through the activation of the melanocortin-1 (MC-1) receptor, which is expressed by melanocytes. Recent evidence suggests that the related POMC peptides such as adrenocorticotrophin (ACTH), which is the precursor of alpha-MSH, is also an agonist at the MC-1 receptor. By using immunocytochemistry, we confirmed the presence of alpha-MSH in human skin where staining was evident in keratinocytes and especially strong in melanocytes and possibly Langerhans cells. ACTH was also present and tended to show the strongest reaction in differentiated keratinocytes. Immunostaining was also observed for the prohormone convertases, PC1 and PC2, which are involved in the formation of ACTH and its cleavage to alpha-MSH, respectively. The amounts of immunoreactive ACTH exceeded those of alpha-MSH. Using HPLC we identified for the first time the presence of ACTH1-39, ACTH1-17, ACTH1-10, acetylated ACTH1-10, alpha-MSH, and desacetyl alpha-MSH in epidermis and in cultured keratinocytes. The ability of these peptides to activate the human MC-1 receptor was examined in HEK 293 cells that had been transfected with the receptor. All peptides increased adenylate cyclase in these cells with the following order of potency: ACTH1-17 > alpha-MSH > ACTH1-39 > desacetyl alpha-MSH > acetylated ACTH1-10 > ACTH1-10. ACTH1-17 also increased the dendricity and melanin content of cultured human melanocytes indicating that the peptide was able to activate MC-1 receptors when present in their normal location. However, as found with alpha-MSH, not all cultures were responsive and, as we have previously suggested, we suspect that this was the result of changes at the MC-1 receptor. Nevertheless, it would appear that ACTH peptides can serve as natural ligands of the MC-1 receptor on human melanocytes and their presence in the skin suggests that, together with alpha-MSH, they may have a role in the regulation of human melanocytes. PMID- 9359625 TI - Agouti protein inhibits the production of eumelanin and phaeomelanin in the presence and absence of alpha-melanocyte stimulating hormone. AB - Melanocytes synthesise two types of melanin: the brown-black eumelanin and the red-yellow phaeomelanin. In mice, the relative proportions of these two melanins are regulated by alpha-MSH, which preferentially increases the synthesis of eumelanin and by the Agouti protein (AP), the expression of which correlates with the growth of yellow phaeomelanin-containing hair. It has been proposed that AP acts by antagonizing the action of alpha-MSH at the MC1 receptor, although it has been suggested that it may also act independently of alpha-MSH. In the present study we show that AP inhibits melanogenesis in B16F1 melanoma cells in the presence and absence of alpha-MSH and also causes dose-related decreases in the synthesis of both eumelanin and phaeomelanin. In the presence of alpha-MSH AP had a greater effect on eumelanin production and this is consistent with an antagonistic action at the MC1 receptor. In the absence of alpha-MSH however, AP produced similar reductions in the synthesis of both melanins. These changes were not seen in B16G4F cells which lack the MC1 receptor, suggesting that even in the absence of alpha-MSH AP acts at the MC1 receptor. How this action is mediated at the intracellular level is not yet clear, although it appears to be associated with a decrease in tyrosinase activity. PMID- 9359626 TI - Establishment of a murine model for metastasis of cytokine-producing tumor to the brain. AB - The A375 cell line, derived from human malignant melanoma, has characteristics of interleukin-6 (IL-6) production. By using this cell line, we have investigated a murine metastasis model of IL-6-producing tumors to the brain by injecting A375 cells directly into the left cardiac ventricle. Nude mice were anesthetized with intraperitoneal injection of pentobarbital sodium. Next, A375 cells suspended in phosphate-buffered saline (PBS) were injected into the left cardiac ventricle of mice. An intracardiac injection of 10(5) cells developed tumor colonies in the brain after 4 to 6 weeks. Metastatic cells were found in every lobe of the brain. An immunocytochemical study revealed IL-6 production by A375 cells at the metastatic sites in the brain. By the transfection of genes encoding proteins into A375 cells, a novel model of protein expression in the brain in vivo could be constructed. Our system does not require great skill. Our experimental model will facilitate future studies of the local effects of proteins in the brain. PMID- 9359627 TI - A periodical change of mercury in the coral reef, Heliofungia species obtained from the Okinawa Sea. PMID- 9359629 TI - Acetaldehyde depresses shortening and intracellular Ca2+ transients in adult rat ventricular myocytes. AB - Acetaldehyde (ACA), an ethanol metabolite, exerts both stimulatory and depressive effects on isolated myocardial tissue, but its impact on individual cardiac myocytes is unknown. The purpose of this study was to determine whether ACA induced myocardial depression is due to an intrinsic alteration of the contractile properties of heart at the cellular level. Mechanical properties of adult rat ventricular myocytes were evaluated using a video edge-detection system. Myocytes were electrically stimulated to contract at 0.5 Hz under isotonic conditions in a physiological buffer containing 1 mM CaCl2. Contractile properties analyzed include: peak twitch amplitude (PTA), time-to-PTA (TPT), time to-relengthening (TR90) and maximal velocities of shortening and relengthening (+/-dL/dt). Ca2+ transients were measured as fura-2 fluorescence intensity (FFI) changes. ACA (1-30 mM) disproportionately depressed PTA and FFI in a dose dependent manner, with maximal inhibitions of 57 and 19%, respectively. Neither the durations nor maximal velocities of shortening and relengthening were affected by ACA. The depression of cell shortening by ACA was either attenuated or blocked by BayK 8644 or elevated extracellular Ca2+ (2.7 mM). In addition, ACA also reduced caffeine-induced FFI changes. These results suggest that ACA-induced myocardial depression in multicellular preparations is due to an intrinsic action on individual myocytes. The mechanism underlying ACA-induced myocardial depression may be due, in part, to either reduced Ca2+ entry through voltage dependent Ca2+ channels and/or depression of sarcoplasmic reticular Ca2+ release. PMID- 9359628 TI - Water soluble free radicals as biologically responsive agents in electron paramagnetic resonance imaging. AB - Electron paramagnetic resonance imaging (EPRI) is currently being developed at frequencies between 200 MHz and 2 GHz. EPRI can map the in vivo distribution of paramagnetic species, such as water soluble free radicals; nitroxide free radicals are commonly used. EPR images reflect the complexity of metabolic actions on the exogenous delivered nitroxides. Their reduction rate in vivo is affected by parameters such as oxygen concentration, pH and biodistribution. This paper illustrates the main features of low frequency EPRI and reconstruction techniques. Examples of EPR imaging, such as two-dimensional (2D) spatial mapping of the distribution of a nitroxide free radical in phantoms and in whole rat, are given. PMID- 9359630 TI - Hexosaminidase in Trichinella spiralis is a single protein with alpha- and beta subunits catalytic activities. AB - Beta-N-acetylhexosaminidase is expressed as a single protein in Trichinella spiralis and has catalytic properties similar to the alpha- and beta-subunits of human and mouse isoenzymes A and B. It can hydrolyze the artificial substrates, 4 methylumbelliferyl-beta-D-glucosamine and 4-methylumbelliferyl-beta-D-glucosamine 6-sulphate which are respectively hydrolyzed by the beta- and alpha-subunits. The enzyme is thermostable, has a basic isoelectric point, and thus is similar to the B isoenzyme. Northern blotting experiments indicate that the enzyme is encoded by a single gene. Hexosaminidase from Trichinella spiralis shows that the substrate specificities of alpha- and beta-subunits precede the duplication of their genes. PMID- 9359631 TI - How to correct for errors in mRNA quantitation by competitive PCR due to heteroduplex formation of amplification products. AB - The Q-RT-PCR method for determining mRNA levels is an internally-controlled quantitative reverse transcriptase linked polymerase chain reaction to assess gene activity by monitoring the accumulated stable transcript level, in a given sample of extracted total RNA. Internal and competitive standards are used for mRNA titration because of the exponential nature of PCR; possible variations in RT efficiency are corrected for by the use of riboprobe RNA standards. A renin mRNA assay has been optimized by altering PCR primer parameters. Q-RT-PCR suffers from the occurrence of heteroduplex DNA formation between amplification products from homologous standard and target mRNA. Co-migration of various PCR products is shown to occur on neutral agarose gels, and this will lead to serious errors in mRNA determinations. Adjustment of the standard electrophoresis conditions is required for absolute mRNA quantitation by Q-RT-PCR. PMID- 9359632 TI - The role of cytoskeletal elements in the two-phase denucleation process of mammalian erythroblasts in vitro observed by laser confocal scanning microscope. AB - The cytoskeletal elements in the denucleation processes were observed using immunofluorescence and laser confocal scanning microscopy in the Friend virus (FVA) infected splenic erythroblasts of BALB/c mice. When cultured in the presence of erythropoietin (EPO), it was shown that the synchronized erythroid precursor cells proceeded to an autonomous nuclear extrusion when the three types of cytoskeletal elements were observed contributing to different phases of that process. The vimentin intermediate filament (IF) was shown as the nuclear anchorage elements with binding sites anchored from the nuclear lamina to the center as well as to the plasma membrane periphery. A dense perinuclear layer of vimentin fluorescence in erythroblasts was observable during the periods of 12, 24 and 36 hrs. in vitro culture. The amount of vimentin IF per cell was higher than that of tubulin and F-actin at 12-24 hrs. culture, but the vimentin filaments were observed to brake down and decreased steadily when the cells became differentiated into late erythroblasts at 36-48 hrs. Such an attenuation of vimentin filaments may facilitate the eccentric movement of the nucleus which can be regarded as the initial step (phase) of denucleation. The fluorescent intensity of tubulin and actin exhibited a significant rise and aggregated between the extruding nucleus and the incipient reticulocyte prior to and during the processes of denucleation, what indicated that the actin filaments and microtubules may play roles in the second phase of the denucleation process, or final commitment of enucleation. The erythroid differentiation-denucleation factor (EDDF), as an intrinsic factor, involved in the denucleation events, was also discussed. PMID- 9359633 TI - Modulation of proliferative activity and amino acid transport in chick embryo hepatocytes by EGF and retinoic acid. AB - In this work, the proliferative activity of chick embryo hepatocytes and its regulation by factors affecting cell growth, such as epidermal growth factor (EGF) and retinoic acid, were studied. The transport of nutritional molecules, like amino acids, was also investigated; in particular, the uptake mediated by the Na+ -dependent system A, known to be under hormonal and growth factor control. Moreover, some steps in the transduction pathway of mitogenic signal were analyzed, both for EGF and retinoic acid treatment. Results obtained suggest that chick embryo hepatocytes growth in an exclusive serum dependent way shows an early sensibility to EGF stimulation as regards to the proliferative activity and amino acid transport, responds to retinoic acid treatment and lack of contact inhibition. Moreover, as far as the signal transduction is concerned, at early stages, they do not seem to be able to utilize the same signal molecules as observed in adult life. PMID- 9359634 TI - In vivo sensitivity of murine haemopoietic progenitor cell populations to mixed gamma-rays - neutron irradiation at different gamma/n ratios. AB - Changes in mice haemopoietic cellular populations and in the radiosensitivity of CFU-C and BFUe progenitors cells were determined in vivo for mixed field radiations composed of a gamma-ray component and a neutron component. Five Dgamma/Dtotal ratios (gamma-rays over total dose ratios, quoted as tau in this report) were obtained (tau = 0.95, 0.83, 0.67, 0.33 and 0.09). Myelogram changes were enlarged with the increase of the neutron component. Radiosensitivity of the two progenitor cell lineages were increased with lower tau values (excess of neutrons). The radiosensitivity of haemopoietic progenitor cells exposed in vivo varies with the ratio of the high- and low-LET components in the mixture. The D0 value varied from 3.3 +/- 0.22 to 0.85 +/- 0.04 Gy with the decrease of tau for CFU-C and from 2.08 +/- 0.22 to 0.64 +/- 0.07 Gy for BFUe. The obtained relative biologic efficiency (RBE) varied from 1.2 +/- 0.08 to 4.7 +/- 0.24 for CFU-C and from 1.1 +/- 0.1 to 3.6 +/- 0.16 for the BFUe. The relation between RBE and tau could be somewhat non-linear for CFU-C and seems to be close to linear for BFUe. The higher is the neutron component, the higher is the radiosensitivity. These results indicate that variations of the quality of the mixed field in the haemopoietic local territory are of great importance in terms of radiation damage and cell killing as well as in terms of the ability to restore the haemopoietic system. PMID- 9359635 TI - Expression of myotonic dystrophy protein kinase gene during in vivo and in vitro mouse myogenesis. AB - Myotonic dystrophy (DM) is an autosomal dominant neuromuscular disorder characterized by a great variability in its clinical manifestations. The mutational basis underlying DM consists of an unstable (CTG)n trinucleotide repeat in the 3' untranslated region of the myotonic dystrophy protein kinase gene (DMPK). Conflicting results on DMPK gene expression in congenitally affected infants (CDM) have been published. Moreover, the prominence of satellite cells seen in muscle of CDM infants supports the notion that the congenital form is associated with an arrest in muscle development and suggests a role for the DMPK gene during differentiation and maturation of muscle. In order to clarify these findings, a comparative study of DMPK and myogenic factor mRNA levels was performed in developing mouse muscle tissues and cultured muscle cells at different developmental stages. Results show that DMPK gene expression is upregulated at a late stage of muscular development. This upregulation does not seem to depend on a given muscle specific bHLH factor. PMID- 9359636 TI - A cytotoxic factor for glial cells: a new avenue of research for multiple sclerosis? AB - A novel retrovirus, provisionally called Multiple Sclerosis RetroVirus (MSRV), was recently described in multiple sclerosis (MS). We report here that monocyte/macrophage culture supernatants from MS patients containing reverse transcriptase activity secrete a cytotoxin which induces death of primary mouse cortical glial cells. This cytotoxin, which was also found in MS cerebrospinal fluid, specifically causes death of mouse immortalized astrocytes and oligodendrocytes in vitro and seems to be associated to MSRV-specific RNA. This toxic factor, called gliotoxin, is present only in active cases of MS and is a stable glycosylated protein of 17 kDa, in CSF as well as in monocyte/macrophage culture supernatants. Since this gliotoxin is highly toxic for glial cells, it may represent an initial pathogenic factor, leading to the neuropathological features of MS, like blood brain barrier disruption and demyelination. PMID- 9359637 TI - Insulin-like growth factor-binding protein-1 in human follicular fluid: a marker for oocyte maturation. AB - In order to investigate the role of insulin-like growth factors (IGFs) in human ovulation, we evaluated the concentrations of IGF-binding proteins (IGFBPs) in human follicular fluid (FF). The concentrations of IGFBP-1 in the FFs of 15 women undergoing in vitro fertilization and embryo transfer were measured and related to those of 17beta-estradiol (E2), progesterone and androstenedione in the FFs. IGFBP-1 levels in the FFs were positively correlated with those of E2 and progesterone. No correlation was found between the IGFBP-1 and androstenedione levels in FFs. The concentrations of IGFBP-1 were significantly increased in the FFs which contained mature oocytes compared with those of immature oocytes, whereas IGFBP-3 in FFs tended to decrease as oocytes matured. It is suggested that IGFs may play important roles in human preovulatory processes, and that IGFBP-1 may be a valuable biochemical marker in the evaluation of oocytes. PMID- 9359638 TI - Direct intraperitoneal insemination compared to intrauterine insemination in superovulated cycles: a randomized cross-over study. AB - The aim of this randomized cross-over study was to determine whether direct intraperitoneal insemination (DIPI) is superior to intrauterine insemination (IUI) in hyperstimulated cycles. The treatment cycles were stimulated with either clomiphene citrate and human menopausal gonadotrophins, or buserelin and human menopausal gonadotrophins. 207 subfertile couples with a cervical factor, a male factor, a combined cervical and male factor, or an unexplained subfertility were randomly assigned to the first treatment cycle. IUI and DIPI were performed in alternate cycles to a maximum of 6 cycles per couple. Every treatment cycle was followed by a nontreatment cycle. The pregnancy rate per completed cycle was 24% for IUI and 16% for DIPI (p = 0.018), whereas the cumulative pregnancy rates for IUI and DIPI were 53 and 40%, respectively (p = 0.002). There were no significant differences between pregnancy rates for IUI and DIPI in the different categories of subfertility. We conclude that DIPI does not offer better pregnancy chances than IUI in superovulated cycles. PMID- 9359639 TI - Endogenous nitric oxide attenuates ethanol-induced vasoconstriction in the human placenta. AB - The purpose of this study was to clarify the role of endogenous nitric oxide and prostanoids in ethanol-induced perturbation of microcirculation in perfused human placenta. Infusion of ethanol into chorionic plate vessels at 10-65 mM increases perfusion pressure in a concentration-dependent fashion, and is an indicator of fetal-placental vasoconstriction. Simultaneous infusion of N(omega)-nitro-L arginine, methylene blue and endothelial cell removal significantly enhances the ethanol-induced increase in perfusion pressure. In contrast, sodium nitroprusside attenuates this effect. Indomethacin did not significantly modify the ethanol induced response. In conclusion, inhibition of the action of endogenous nitric oxide is associated with an increase in fetal-placental vasoconstriction. These results suggest that endogenous nitric oxide acts as a vasodilator that reduces ethanol-induced vasoconstriction, thus improving microcirculation, and leads to decreased placental damage. PMID- 9359640 TI - Anxiety of infertile women undergoing IVF-ET: relation to the grief process. AB - The psychological state of 102 infertile women undergoing in vitro fertilization and embryo transfer was investigated. The levels of anxiety were measured by the state trait anxiety inventory and the manifest anxiety scale. The process of accepting infertility and attitudes toward treatment were explored by a semistructured interview. The mean score of state anxiety for the subjects was 50.0 which was considerably higher than the standard score of 42 for the general Japanese females. Women undergoing in vitro fertilization and embryo transfer with higher levels of anxiety remain in the introversive stage of the grief process, have a more positive attitude toward treatment, a more pessimistic outlook on the possibility of successful pregnancy, and feel more agitation. PMID- 9359641 TI - Association of blood loss during delivery to B-hemoglobin. AB - OBJECTIVE: To study the association between blood loss during delivery and B hemoglobin before and after delivery (3 days and 10 weeks, respectively). MATERIALS AND METHODS: Information on blood loss for 693 women, 93.9% of all parturients during the study period, was extracted from the original medical records. B-hemoglobin was analyzed at the antenatal maternal health care unit at the last visit before delivery, in capillary samples taken on the ward on the 3rd day, or 10 weeks after delivery. Associations were estimated with Pearson's parametric correlation coefficient. RESULTS: Altogether, 31.3% of all parturients had a higher B-hemoglobin value on the 3rd day after delivery than on the last visit to the maternal health care unit before delivery; the mean intrapartum blood loss was 375 ml (range 100-2,200 ml). The correlation coefficient between B hemoglobin on the 3rd day after delivery and blood loss was r = -0.53. When the last value recorded at the antenatal maternal health care unit was included in the analysis, the correlation coefficient remained virtually unchanged (r = 0.52). Only 14 (blood loss < or = 600 ml), or 11% (blood loss >600 ml) of the variation in the B-hemoglobin was accounted for by the amount of blood loss. No significant correlation coefficient was evident for B-hemoglobin 10 weeks after delivery vis-a-vis blood loss, irrespective of whether iron supplementation was administered (r = 0.01) or not (r = 0.17). CONCLUSION: The weak association between intrapartum blood loss and B-hemoglobin suggests that the value of B hemoglobin determined after delivery may be less indicative than previously thought. This conclusion is strengthened by the fact that only a minor part (< or = 14%) of the variation in B-hemoglobin on the 3rd day after delivery is explained by the amount of blood lost. PMID- 9359642 TI - Preterm singleton breech in North Jordan: vaginal versus abdominal delivery. AB - The safety of vaginal birth for singleton preterm breech has not often been addressed before. We retrospectively compared the perinatal outcome of two groups of preterm breech delivery. Sixty-six patients delivered vaginally and 32 delivered abdominally between 26 and 36 completed weeks. Vaginal delivery was allowed under the same protocol for singleton breech delivery at term. Both groups had similar maternal characteristics. Intergroup differences in early neonatal outcome, as measured by Apgar score, were not significant. Intrapartum and early neonatal deaths in vaginal and cesarean delivery were compared. There was no significant difference in intrapartum death and early neonatal mortality between those who delivered vaginally and those who delivered by cesarean section (16.6 vs. 15.6%). So even with optimum neonatal care facilities, cesarean section does not offer any advantage over vaginal delivery in a developing country. This study does not advocate the routine use of cesarean section for delivering preterm breech fetuses. PMID- 9359643 TI - Plasma catecholamines and Doppler-derived cardiac time intervals in vaginally and cesarean delivered neonates. AB - OBJECTIVE: Our purpose was to compare Doppler-derived cardiac time intervals and cord plasma levels of catecholamines in vaginally and cesarean delivered neonates. METHODS: Doppler echocardiographic assessments of fetal and neonatal cardiac time intervals were made to determine differences in circulatory changes in 8 neonates delivered vaginally and 12 delivered by elective cesarean section. Aortic and pulmonary acceleration time (AT), ejection time (ET), and the AT/ET ratio in systole were assessed at various time points from antenatal to 72 h after delivery. Umbilical artery blood gas and acid-base values, and cord blood concentrations of epinephrine and norepinephrine were also measured. RESULTS: Umbilical artery blood pH in the cesarean section group was significantly lower than that in the vaginal delivery group (p < 0.05). Base deficit in umbilical artery blood and cord blood norepinephrine in cesarean delivered neonates were significantly higher than those in vaginally delivered neonates, respectively (p < 0.05). There were no significant changes in aortic cardiac time intervals between the 2 groups at any of the various time points. However, pulmonary AT and the AT/ET ratio were significantly lower in the normal vaginal delivery group than those in the cesarean section group 6 h after delivery (p < 0.05). CONCLUSIONS: The lower AT and AT/ET ratio of the pulmonary artery in vaginally delivered neonates may reflect an increase in pulmonary vascular resistance, indicating transient pulmonary hypertension. The findings were the reverse of what might be expected from an elevated pulmonary vascular resistance and transient pulmonary hypertension in cesarean delivered neonates. PMID- 9359644 TI - Nitric oxide synthase activity in umbilical and placental vascular tissue of gestational diabetic pregnancies. AB - OBJECTIVE: Nitric oxide (NO) synthase activity in the fetal-placental vasculature of gestational diabetic pregnancies was compared to non-diabetic controls. METHODS: Placentae were collected from non-diabetic deliveries (n = 5) and from patients with gestational diabetes (n = 8). Umbilical cord and chorionic plate arteries and veins and stem villous vessels were quickly dissected out, cleaned of contaminating tissue, snap frozen in liquid nitrogen and stored at -80 degrees C. NO synthase activity was measured in the homogenate of these vessels by the arginine to citrulline conversion assay using 5 microM 3H-L-arginine with 1 mM NADPH, 100 microM free calcium, 50 units/ml calmodulin, 10 microM tetrahydrobiopterin and 2 microM flavin adenine dinucleotide with 45-min incubation at 27 degrees C. Enzyme activity was expressed as picomoles of 3H-L citrulline formed per milligram of protein per minute. RESULTS: No significant differences in NO synthase activity were found between non-diabetic and gestational diabetics for umbilical cord artery (0.58 +/- 0.22 vs. 0.19 +/- 0.06), cord vein (0.39 +/- 0.21 vs. 0.07 +/- 0.03), chorionic plate artery (0.32 +/- 0.14 vs. 0.26 +/- 0.19) or vein (0.41 +/- 0.15 vs. 0.22 +/- 0.06), respectively (mean +/- SEM). Significantly greater activity was found in stem villous vessels of non-diabetic placentae (5.64 +/- 2.0) compared to those of gestational diabetic placentae (0.48 +/- 0.19; p < 0.01). CONCLUSION: The reduced blood flow and increased vascular resistance observed in diabetic pregnancies may be due to decreased NO synthesis and activity in the stem villous vessels which are the major determinants of resistance in the fetal-placental vasculature. PMID- 9359645 TI - Female and male sterilization and refertilization in Sweden. AB - In Sweden, approximately 1,500 men and 7,000 women are sterilized yearly. The frequency varies considerably between regions. The reasons for these differences are not fully known. More women than men are sterilized in Sweden, but a higher proportion of sterilized men undergo refertilization. The object of the study was to analyze counselling routines prior to sterilization, presterilization waiting time, the number of refertilizations performed, the attitudes of the operating departments towards refertilization, and the possible relation between frequency of refertilization and incidence of sterilization. A questionnaire was mailed to all units performing refertilizations and/or sterilizations of mean and women in Sweden (n = 161). The response was 93%. Counselling routines and waiting times before sterilization varied, which influenced the operation's availability. The frequency of refertilizations varied considerably between counties from 0.5 to 5.4 per 100,000 inhabitants. There was no correlation between frequency of sterilizations and refertilizations. The rate of refertilization appears to be determined mostly by the local attitudes of the health authority. The Swedish sterilization law has had a heterogeneous impact in the country. PMID- 9359646 TI - Sonographic measurement of endometrial thickness as a predictor of vaginal bleeding in women using continuous combined hormone replacement therapy. AB - The development of unpredictable vaginal bleeding is one of the main reasons for the discontinuation of treatment in users of continuous combined hormone replacement therapy (HRT). There is no accurate method of predicting which women will develop bleeding on this regimen. The aim of this study was to determine whether sonographic assessment of the endometrium could be used as a predictor of bleeding after the commencement of this treatment. Sonographic measurements of the endometrium were performed before treatment and then every 3 months for 1 year. Measurements of the endometrium were taken in both the transverse and sagittal planes and a record of the frequency of bleeding was maintained. Pretreatment measurements tended to be greater in those who developed bleeding within 1 month of the commencement of treatment, but the difference in measurements between those who developed bleeding and those who did not was not statistically significant. Endometrial thickness tended to be greater at 3, 6, 9 and 12 months of treatment in those who had bleeding compared with those who did not, but again the difference was not statistically significant. It is concluded that whilst bleeding on continuous combined HRT is more common in women with a thicker endometrium, sonographic assessment of the endometrium is not an accurate enough predictor of bleeding to be used in this clinical setting. PMID- 9359648 TI - Continuous local injection of carboplatin into the uterine cervix for cervical and endometrial cancer: preliminary study. AB - Platinum compounds are thought to be concentration- and time-dependent, but intravenous (i.v.) administration does not afford prolonged high platinum concentration in tumor tissue. In order to examine the influence of long-term local continuous (LC) injection of carboplatin, a pharmacokinetic study was performed. Twenty-six patients with uterine cancer were included. I.v. administration (11 patients): carboplatin (210 mg) was given i.v. and samples of target tissue were obtained at operation about 2 or 24 h after administration. LC administration (15 patients): the 21-gauge needle was implanted at the uterine cervix, and carboplatin was injected continuously (30 mg/day) for 3, 7 or 14 days using an external pump. The tissue platinum concentration was measured in the pelvic organs. The mean platinum levels at the cervix and vaginal wall in the LC (7 days) group were higher than those in the i.v. (2 h) group (p < 0.01). With LC injection, sustained platinum levels were maintained in the pelvic organs for a long time, with very few side effects. LC injection may be advantageous on the basis of pharmacokinetics. PMID- 9359647 TI - Cervical polyp as risk factor for hysteroscopically diagnosed endometrial polyps. AB - A case-control study was conducted searching through 590 diagnostic hysteroscopies in order to identify potential risk factors for endometrial polyps. Case (hysteroscopically positive for endometrial polyps) and control groups were compared for age, parity, cervical polyp, menopausal status, smoking and current use of contraceptive pills. A higher prevalence of endometrial polyps was found among women with a cervical polyp (26.9%) compared to those without a cervical polyp (7.1%, chi(2) = 27.52, p < 0.001). Increased risk of endometrial polyp was found to be associated with age (odds ratio = 1.75 every 10 years, p < 0.001), cervical polyp (odds ratio = 4.83, 95% CI = 2.43-9.52), and menopause (odds ratio = 2.03, 95% CI = 1.12-3.69). After multivariate analysis, only age (adjusted odds ratio = 1.90, p = 0.001) and cervical polyp (adjusted odds ratio = 5.42, p < 0.001) were independent variables still associated with increased risk of endometrial polyps. We conclude that age and cervical polyp are strong, independent risk factors for endometrial polyps. PMID- 9359649 TI - High frequency of adenomyosis in postmenopausal breast cancer patients treated with tamoxifen. AB - Pathologic evaluation for adenomyosis in uterine specimens as well as demographic characteristics, health habits and risk factors for endometrial cancer were compared in 28 postmenopausal breast cancer patients with tamoxifen (TAM) treatment and in 11 similar patients without TAM treatment in order to determine the association between postmenopausal TAM exposure and the frequency of adenomyosis. The same comparison was also made between TAM-treated patients with adenomyosis and TAM-treated patients without adenomyosis. Adenomyosis was histologically diagnosed in 53.6% TAM-treated patients and in 18.2% non-TAM patients. Overall, there were no significant statistical differences in all parameters tested between the 2 groups, as well as between the TAM-treated patients with adenomyosis and the TAM-treated patients without adenomyosis. It can be concluded that adenomyosis was significantly more common among postmenopausal breast cancer patients who were treated with TAM as compared to similar patients without TAM treatment (p = 0.0186). This significant high rate of adenomyosis may be attributed to the continuous and unopposed exposure to TAM. It is, however, impossible to predict which postmenopausal breast cancer patient will develop adenomyosis after treatment with TAM. PMID- 9359650 TI - Hydranencephaly with renal dysgenesis: a coincidental finding? Case report with review of the literature. AB - We present the case of a female fetus aborted in the 20th week of gestation due to severely dysplastic kidneys, anhydramnios and hydranencephalus. The combination of these malformations is extremely rare, resulting in only 4 cases described so far. Our case is the first ever presented in a female showing polycystic-dysplastic kidneys. Multiple multinucleated neurons were a remarkable finding in the remnants of the brain. The possibility of an underlying genetic disorder is discussed, together with a brief review of the literature to date. PMID- 9359651 TI - Successful treatment of a high-risk choriocarcinoma in Figo stage IV (WHO score 12). AB - In the present report, a case of high-risk choriocarcinoma (Figo stage IV, WHO score 12) with persisting vaginal and urethral bleedings is described. In addition to a course of multiagent chemotherapy, arterial embolization of both iliac vessels using Gianturco coils was carried out. This caused a dramatic improvement of the patient's general condition. A subsequently performed laparatomy with extirpation of the uterus and both adnexa finally led to remission. PMID- 9359653 TI - Silent infection of macaques inoculated with low-dose SIVmac251/spl. PMID- 9359652 TI - Central nervous system involvement secondary to metastatic mixed mullerian tumor of the uterus. AB - The central nervous system is traditionally considered an uncommon site for metastatic disease from female genital tract tumors. We report the case of a 48 year-old woman with malignant mixed mullerian tumor of the uterus, who developed spinal cord compression by epidural metastasis a few days after the diagnosis of the uterine malignancy. Emergency decompressive laminectomy was performed and a good recovery of the neurological function was achieved. In the following days, while submitted to extensive staging for the uterine malignancy, the patient complained of headache, confusion and visual disturbance. CT scan revealed multiple brain metastases. No other site of metastatic disease could be detected. The patient refused any further treatment and died 1 month later from progressive cerebral disease. Attention should be paid to the possibility of unusual distant metastases associated to uterine sarcoma in order to treat these patients promptly. PMID- 9359654 TI - HIV type 1 subtypes, coreceptor usage, and CCR5 polymorphism. AB - Identification of the chemokine receptors CCR5 and CXCR4 as the major coreceptors for HIV-1 entry has greatly assisted our understanding of HIV-1 pathogenesis, transmission, and tropism. However, most of our current knowledge on coreceptor usage comes from studies using HIV-1 strains or env genes derived from the genetic subtype B predominant in North America and western Europe. In this report, the coreceptor usage of 20 primary viral isolates representative of genetic subtypes A, B, C, D, E, and group O was examined. Thirty-nine full-length CCR5 sequences from individuals of diverse geographic origins were also obtained to examine the possible effect of CCR5 polymorphism on HIV-1 subtype distribution. Our results indicate that (1) CCR5 and CXCR4 serve as the two major coreceptors for viruses belonging to HIV-1 subtypes A, B, C, D, E, and group O, whereas other chemokine receptors such as CCR2b and CCR3 play only a minor role in facilitating viral entry into stimulated PBMCs; (2) the coreceptor usage is determined by the viral phenotype rather than its genotype because all NSI strains, irrespective of their subtype classification, utilize CCR5, whereas all SI strains are able to use CXCR4; and (3) there is no geographic clustering of CCR5 polymorphism in different ethnic populations, suggesting that CCR5 diversity is not the underlying explanation for differences in the spread of different HIV 1 subtypes. Therefore, the uneven worldwide distribution of HIV-1 subtypes is more likely the result of stochastic dissemination. PMID- 9359655 TI - C-C chemokine RANTES and HIV long terminal repeat-driven gene expression. AB - The C-C chemokines RANTES, MIP-1alpha, and MIP-1beta have been characterized as constituents of an HIV- and SIV-suppressive factor released by CD8+ cells. Furthermore, it has been demonstrated that chemokine receptors cooperate in HIV entry. However, these proteins are also known to have an effect on multiple intracellular signaling cascades that may affect the process of transcription. In the present study we demonstrate that treatment of CD4+ T cells with these chemokines or with cell supernatants from HTLV-I-immortalized CD8+ T cells results in significant reduction in the abundance of HIV-1-specific RNA as analyzed by Northern blot hybridization. To examine the possibility that such suppressive factors may inhibit HIV RNA transcription, we studied the effect of RANTES, the most effective HIV-suppressive chemokine, on basal and Tat-induced HIV-directed LTR expression of a reporter gene. Neither recombinant RANTES nor conditioned medium from CD8+ cells significantly altered HIV-1 LTR-directed chloramphenicol acetyltransferase expression in either transiently or stably transfected CD4+ T cell lines, either in the presence or in the absence of Tat. These results suggest that C-C chemokines do not inhibit viral RNA transcription. PMID- 9359656 TI - Low frequency of CCR5delta32 allele among Greeks in Cyprus. PMID- 9359657 TI - Early postinfection antiviral treatment reduces viral load and prevents CD4+ cell decline in HIV type 2-infected macaques. AB - Reports of significant reductions in plasma viral load by anti-HIV drugs have raised the possibility that antiviral therapy, if initiated sufficiently early, may result in sustained control of infection and prolonged clinical benefits. We evaluated the effects of intervention coincident with infection using an antiviral nucleoside, d4T, in Macaca nemestrina infected with a highly pathogenic isolate of HIV-2 (HIV-2[287]). Infection with this virus reproducibly results in high viremia and rapid CD4+ cell depletion, allowing a sensitive measurement of the treatment effect on viral load and clinical outcome. Compared to the control group, d4T-treated macaques showed significantly lower (2-3 log10) plasma- and cell-associated viral load. No CD4+ cell decline was observed in the treatment group while on therapy with d4T whereas CD4+ cells of control macaques declined from a preinfection mean of 32% of PBMCs to below 10%. Notably, when d4T treatment was withdrawn after 16 weeks, five of the six macaques continued to control viral load and have maintained normal CD4+ cell level for more than a year. These results demonstrate that early antiviral intervention, even of a limited duration, may constitute an important strategy against lentiviral-induced disease. PMID- 9359659 TI - Antiviral potency of drug-gene therapy combinations against human immunodeficiency virus type 1. AB - Gene therapy for the treatment of human immunodeficiency virus type 1 (HIV-1) infection using intracellular immunization strategies is currently being tested in clinical trials. With the continuing development of potent antiretroviral drugs (e.g., reverse transcriptase [RT] and protease [PR] inhibitors), it is likely that HIV-1 gene therapy will be applied to humans concurrently receiving such antiretroviral medication. In this study, we assessed the in vitro antiviral efficacy of two gene therapy strategies (trans-dominant RevM10, Gag antisense RNA) in combination with clinically relevant RT (AZT, ddC) or PR (indinavir) inhibitors. Retrovirally transduced, human T cell lines expressing antiviral gene constructs were inoculated with high doses of HIV-1HXB3 in the presence or absence of inhibitors. The combination of RevM10 or Gag antisense RNA with antiviral drugs inhibited HIV-1 replication 10-fold more effectively than the single antiviral drug regimen alone. More importantly, we also addressed whether gene therapy strategies are effective against drug-resistant HIV-1 isolates. Both the RevM10 and Gag antisense RNA strategies showed antiviral efficacy against several RT inhibitor-resistant HIV-1 isolates equivalent to their inhibition of HIV-1HXB3 replication. In summary, our data demonstrate the greater than additive antiviral efficacy of gene therapy strategies and RT or PR inhibitors, and that gene therapy approaches are effective against drug-resistant HIV-1 viral isolates. PMID- 9359658 TI - Course of specific T lymphocyte cytotoxicity, plasma and cellular viral loads, and neutralizing antibody titers in 17 recently seroconverted HIV type 1-infected patients. AB - Relationships were sought between specific anti-HIV cytotoxic T lymphocyte (CTL) responses (against structural and regulatory proteins of the HIV-1 LAI isolate) and plasma and cellular viral loads (VLs) in 17 recently HIV-1-infected patients including 3 displaying asymptomatic primary infection (PI) followed up for 12 months. Plasma VL was correlated directly with CD8 counts and inversely with CD4 counts. Cytotoxic reactions were observed in all patients and directed mainly against structural proteins. The earliest CTL responses were against Gag and Env proteins detected in 87 and 75% of the subjects, respectively, within the first month following PI. Anti-Env and Gag cytotoxic responses were inversely correlated with the plasma VL. Reactions against the pol gene products were thought to be either less involved in or less efficient for the initial decrease of viremia. Responses against regulatory gene products were weak and variable, apart from Nef, which was recognized by half of the subjects. Neutralizing antibodies were not detected before month 3, and were found only in six patients at subsequent times. Two of three patients with asymptomatic PI had a low viral burden and either a delayed response or one limited to a few protein CTL responses, suggesting that the magnitude of the CTL response depends on the initial plasma VL. The third patient displayed viral and CTL parameters identical to those of the patients with symptomatic PI. However, two subjects with symptomatic PI exhibited similarly low plasma VL and moderate CTL responses. Overall, the results suggest that the CTL response may not be the sole factor controlling viremia. PMID- 9359661 TI - Lymphocyte trafficking and HIV infection of human lymphoid tissue in a rotating wall vessel bioreactor. AB - The pathogenesis of HIV infection involves a complex interplay between both the infected and noninfected cells of human lymphoid tissue, the release of free viral particles, the de novo infection of cells, and the recirculatory trafficking of peripheral blood lymphocytes. To develop an in vitro model for studying these various aspects of HIV pathogenesis we have utilized blocks of surgically excised human tonsils and a rotating wall vessel (RWV) cell culture system. Here we show that (1) fragments of the surgically excised human lymphoid tissue remain viable and retain their gross cytoarchitecture for at least 3 weeks when cultured in the RWV system; (2) such lymphoid tissue gradually shows a loss of both T and B cells to the surrounding growth medium; however, this cellular migration is reversible as demonstrated by repopulation of the tissue by labeled cells from the growth medium; (3) this cellular migration may be partially or completely inhibited by embedding the blocks of lymphoid tissue in either a collagen or agarose gel matrix; these embedded tissue blocks retain most of the basic elements of a normal lymphoid cytoarchitecture; and (4) both embedded and nonembedded RWV-cultured blocks of human lymphoid tissue are capable of productive infection by HIV-1 of at least three various strains of different tropism and phenotype, as shown by an increase in both p24 antigen levels and free virus in the culture medium, and by the demonstration of HIV-1 RNA-positive cells inside the tissue identified by in situ hybridization. It is therefore reasonable to suggest that gel-embedded and nonembedded blocks of human lymphoid tissue, cocultured with a suspension of tonsillar lymphocytes in an RWV culture system, constitute a useful model for simulating normal lymphocyte recirculatory traffic and provide a new tool for testing the various aspects of HIV pathogenesis. PMID- 9359660 TI - Combination of a drug targeting the cell with a drug targeting the virus controls human immunodeficiency virus type 1 resistance. AB - Combinations of drugs targeting viral proteins have been used to limit or control drug resistance, which is the most important cause of treatment failure in HIV-1 infected individuals. We suggest an alternative approach, namely to target cellular proteins, which are less prone to mutations than viral proteins. Here we show that simultaneous inhibition of a cellular protein (by hydroxyurea) and a viral protein (by ddI) produces a consistent and sustained suppression of HIV-1 for as long as 40 weeks in the absence of virus rebound. We identified the mechanism to explain this lack of rebound: although the combination of the two drugs did not prevent the emergence of mutant viral strains resistant to didanosine (ddI) in these patients, the mutants were still sensitive to standard doses of ddI in the presence of hydroxyurea. These in vivo results were consistent with our in vitro observations: HIV-1 molecular clones resistant to ddI were rendered sensitive to this drug (at concentrations routinely achievable in vivo) after addition of hydroxyurea. This phenomenon can be explained by the observation that hydroxyurea decreases the level of dATP, the cellular competitor of ddI. A low level of dATP favors the incorporation of ddI, even if the viral reverse transcriptase is resistant to this nucleoside analog. This is a novel mechanism of control of resistance and it explains the efficacy of a treatment that is well tolerated, simple, and inexpensive. PMID- 9359662 TI - Cationic liposomes are a strong adjuvant for a DNA vaccine of human immunodeficiency virus type 1. AB - Liposomes have been widely used to enhance the immune response. In the present investigation, we studied their in vivo immunomodulation of an HIV-1-specific DNA vaccine candidate (pCMV160/REV) constructed with the cytomegalovirus (CMV) promoter-conjugated HIV-1 env and rev DNA plasmids. By immunizing with pCMV160/REV and cationic liposomes through various routes (intramuscular, intraperitoneal, subcutaneous, intradermal, and intranasal), we induced higher levels of both antibody production and delayed-type hypersensitivity (DTH) than by using DNA vaccine alone. The HIV-1-specific cytotoxic T lymphocyte (CTL) activity was observed to be stronger on immunization with the DNA vaccine and cationic liposome combination. The intramuscular, intraperitoneal, and intranasal inoculation routes were more effective in inducing strong DTH and antibody responses than the subcutaneous and intradermal routes. Taken together, these results suggest that cationic liposomes can be highly effective when used with DNA vaccines and administered by various routes. PMID- 9359663 TI - Transcriptional activation of the intercellular adhesion molecule 1 (CD54) gene by human T lymphotropic virus types I and II Tax is mediated through a palindromic response element. AB - In vitro infection of T cells with human T lymphotropic virus types I and II (HTLV-I and HTLV-II) resulted in constitutive expression of ICAM-1. Higher levels of ICAM-1 mRNA were expressed in HTLV-transformed cell lines (MT-2, MoT, C8166) when compared with uninfected T cell lines (A301). We demonstrate that this activation is conferred through a site on the ICAM-1 promoter that is activated in trans by the Tax protein of HTLV-I and HTLV-II. Enhanced promoter activity was detected when the ICAM-1 construct (-1162/+1) was transfected into HTLV-I infected (MT-2), HTLV-II-infected (MoT, AI 1050), or an HTLV-I Tax-only expressing (C8166) cell line as compared to the uninfected T cell line (A3.01). Cotransfection of the uninfected T cell line A3.01 with the ICAM construct along with Tax-I and Tax-II expression plasmid also resulted in increased promoter activity. Furthermore, experiments with deletion constructs of the ICAM-1 promoter region indicated that a region between -88 and -53 bp relative to the transcription start site is sufficient for Tax-inducible CAT expression. This segment includes an 11-bp palindromic segment (TTTCCGGGAAA) that has homology with the IFN-gamma and IL-6 response element. An 11-bp segment containing this regulatory region proved to be sufficient to confer Tax-I and Tax-II inducibility on a heterologous promoter (TK-CAT). Taken together these findings indicate that constitutive expression of ICAM-1 by HTLV-infected cells is influenced by the viral trans-activator protein Tax. This increased expression of ICAM-1 in response to the Tax protein may play an important role in the lymphoproliferation associated with HTLV infection. PMID- 9359666 TI - Learning from scientific misconduct. PMID- 9359664 TI - Diversity of full-length subtype E HIV type 1 env sequences in early seroconvertors from northern Thailand. PMID- 9359665 TI - HIV type 1 envelope sequences from seroconverting patients in Barbados. PMID- 9359667 TI - Italian population yields world's largest twin registry. PMID- 9359668 TI - Clear about cancer therapies. PMID- 9359670 TI - Heparan sulfate and viral tropism. PMID- 9359671 TI - Women in trials: clinical hold or stranglehold? PMID- 9359669 TI - VEGF-nitric oxide reciprocal regulation. PMID- 9359672 TI - The UK endorses DIY genetic screening. PMID- 9359673 TI - Satcher a safe bet for Surgeon General. PMID- 9359675 TI - Indian contract research -- economic necessity or sellout? PMID- 9359676 TI - New lease on life for aging research. PMID- 9359674 TI - "Innovation" in AIDS vaccines. PMID- 9359678 TI - Cancer researchers travel to China. PMID- 9359677 TI - AMA aims for ethical high road following Sunbeam disaster. PMID- 9359680 TI - Repeated transfusion for sickle cell. PMID- 9359679 TI - $34 million neuropeptide deal. PMID- 9359681 TI - Commercial interest in cord blood escalates. PMID- 9359682 TI - Italy encourages return of research scientists. PMID- 9359683 TI - Philanthropic goals. PMID- 9359684 TI - NF-kappaB and bone: the breaking point. PMID- 9359685 TI - A green light for virulence gene expression. PMID- 9359686 TI - To oxidize or not to oxidize? PMID- 9359687 TI - TNF inhibition and sepsis -- sounding a cautionary note. PMID- 9359688 TI - Healing hormones. PMID- 9359689 TI - The ABCs of AMD. PMID- 9359690 TI - The end replication problem: more than one solution. PMID- 9359691 TI - Dimers are a girl's best friend. PMID- 9359692 TI - Polyglutamines, nuclear inclusions and neurodegeneration. PMID- 9359693 TI - Quantitative angiogenesis assays: progress and problems. PMID- 9359694 TI - Estrogen accelerates cutaneous wound healing associated with an increase in TGF beta1 levels. AB - The cellular and molecular mechanisms underlying the effects of aging on human cutaneous wound healing are poorly understood, and the possible role of reproductive hormones in this process has never been investigated. We report that aging in healthy females was associated with a reduced rate of cutaneous wound healing, but an improved quality of scarring both microscopically and macroscopically, and with reduced levels of transforming growth factor-beta1 (TGF beta1) immunostaining and steady-state mRNA in the wound. These age-related changes were reversed by the systemic administration of hormone replacement therapy (HRT). Moreover, ovariectomized young female rodents exhibited a marked delay in repair of acute incisional wounds, which was reversed by the topical application of estrogen. The cellular mechanism underlying these changes appears to involve an estrogen-induced increase in latent TGF-beta1 secretion by dermal fibroblasts. These results suggest that both the rate and quality of wound healing depend on reproductive hormone levels. PMID- 9359695 TI - Expression of truncated utrophin leads to major functional improvements in dystrophin-deficient muscles of mice. AB - Dystrophin-deficient mice (mdx) expressing a truncated (trc) utrophin transgene show amelioration of the dystrophic phenotype. Here we report a multifunctional study demonstrating that trcutrophin expression leads to major improvements of the mechanical performance of muscle (that is, force development, mechanical resistance to forced lengthenings and maximal spontaneous activity) and of the maintenance of the intracellular calcium homeostasis. These are two essential functions of muscle fibers, known to be impaired in mdx mouse muscles and Duchenne muscular dystrophy (DMD) patients. Our results bring strong support to the hypothesis that muscle wasting in dystrophin-deficient DMD patients could be prevented by upregulation of utrophin. PMID- 9359696 TI - Halting angiogenesis suppresses carcinoma cell invasion. AB - The importance of angiogenesis in malignant tumor growth has been interpreted mainly in terms of oxygen and nutrient supply. Here we demonstrate its fundamental role for tumor invasion of malignant human keratinocytes in surface transplants on nude mice. Distinct patterns of angiogenesis and vascular endothelial growth factor receptor-2 (VEGFR-2) expression allowed us to distinguish between benign and malignant cells. Functional inactivation of VEGF R2 by a blocking antibody disrupted ongoing angiogenesis and prevented invasion of malignant cells, without reducing tumor cell proliferation. The reversion of a malignant into a benign phenotype by halting angiogenesis demonstrates a significant function of vascular endothelium for tumor invasion. PMID- 9359697 TI - Apoptosis-associated signaling pathways are required for chemotherapy-mediated female germ cell destruction. AB - Female sterility resulting from oocyte destruction is an unfortunate, and in many cases inevitable, consequence of chemotherapy. We show that unfertilized mouse oocytes exposed to therapeutic levels of the antitumor drug, doxorubicin (DXR), undergo apoptosis; however, fertilized oocytes do not initiate apoptosis, but enter cell-cycle arrest, when treated with DXR. Apoptosis induced by DXR in oocytes is blocked by sphingosine-1-phosphate, an inhibitor of ceramide-promoted cell death. Oocytes from Bax-deficient, but not p53-null, female mice display complete resistance to DXR-induced apoptosis in vivo and in vitro. Pretreatment of oocytes with a specific peptide inhibitor of caspases also abrogates the apoptotic response to DXR. These findings indicate that oocyte destruction caused by chemotherapy can be prevented by manipulation of apoptosis-associated signaling pathways. PMID- 9359698 TI - Antioxidants enhance the cytotoxicity of chemotherapeutic agents in colorectal cancer: a p53-independent induction of p21WAF1/CIP1 via C/EBPbeta. AB - Colorectal cancer (CRC) is the second leading cause of cancer deaths in the United States. Five-fluorouracil (5FU) remains the single most effective treatment for advanced disease, despite a response rate of only 20%. Herein, we show that the antioxidants pyrrolidinedithiocarbamate and vitamin E induce apoptosis in CRC cells. This effect is mediated by induction of p21WAF1/CIP1, a powerful inhibitor of the cell cycle, through a mechanism involving C/EBPbeta (a member of the CCAAT/enhancer binding protein family of transcription factors), independent of p53. Antioxidants significantly enhance CRC tumor growth inhibition by cytotoxic chemotherapy in vitro (5FU and doxorubicin) and in vivo (5FU). Thus, chemotherapeutic agents administered in the presence of antioxidants may provide a novel therapy for colorectal cancer. PMID- 9359699 TI - HIV-1 induction of CD40 on endothelial cells promotes the outgrowth of AIDS associated B-cell lymphomas. AB - Human immunodeficiency virus (HIV)-1 infection is associated with the development of aggressive extranodal B-cell non-Hodgkin's lymphomas. Using microvascular endothelial cell (MVEC)-enriched bone marrow stromal cultures, HIV infection of stromal MVECs from lymphoma patients induced the outgrowth of malignant B cells. MVECs were the only HIV-infected cells in the stroma, and purified brain MVECs also induced a phenotype supportive of neoplastic B-cell attachment and proliferation. HIV infection of MVECs stimulated surface expression of CD40 and allowed preferential induction of the vascular cell adhesion molecule VCAM-1 after CD40 triggering. B-lymphoma cells expressed the CD40 ligand (CD40L), and blocking of CD40-CD40L interactions between HIV-infected MVECs and B-lymphoma cells inhibited B-cell attachment and proliferation. These observations suggest that HIV promotes B-lymphoma cell growth through facilitating attachment of lymphoma cells to HIV-infected MVECs and represent a novel mechanism through which viruses may induce malignancies. PMID- 9359700 TI - HIV-specific mucosal and cellular immunity in HIV-seronegative partners of HIV seropositive individuals. AB - HIV-specific mucosal and cellular immunity was analyzed in heterosexual couples discordant for HIV status in serum and in HIV-unexposed controls. HIV-specific IgA but not IgG was present in urine and vaginal wash samples from HIV-exposed seronegative individuals (ESN), whereas both IgA and IgG were observed in their HIV-seropositive partners; antibodies were not detected in low-risk controls. Envelope protein (Env) peptide-stimulated interleukin-2 (IL-2) production by peripheral blood mononuclear cells (PBMCs) was detected in 9 out of 16 ESNs, 5 out of 16 HIV-infected patients and 1 out of 50 controls. Env peptide-stimulated PBMCs of ESNs produced more IL-2 and less IL-10 compared with those of HIV infected individuals; no differences were observed in chemokine production or in CCR5 expression. These data demonstrate that a compartmentalized immune response to pathogens is possible in humans and raise the possibility of protective roles for cell-mediated immunity and mucosal IgA in HIV-seronegative individuals exposed to HIV. PMID- 9359701 TI - Increased sensitivity of HIV-1 antibody detection. AB - Clinical trial results from 11,344 paired urine and serum samples revealed 1,181 HIV-1-positive individuals confirmed by western blot (WB). There were 25 discrepant samples: 10 were urine enzyme immunoassay (EIA) and WB positive, serum non-reactive and serum WB negative or indeterminate, and 15 were serum EIA and WB positive, urine EIA non-reactive or urine WB negative or indeterminate. Serum samples, HIV-1 antibody WB confirmed, revealed a 99.15% sensitivity (1,171 out of 1,181); urine samples, HIV-1 antibody WB confirmed, showed a 98.73% sensitivity (1,166 out of 1,181). This study demonstrated that neither serum nor urine results alone are as sensitive for HIV-1 antibody detection as combined results of both samples. PMID- 9359702 TI - In vivo evolution of HIV-1 co-receptor usage and sensitivity to chemokine mediated suppression. AB - Following the identification of the C-C chemokines RANTES, MIP-1alpha and MIP 1beta as major human immunodeficiency virus (HIV)-suppressive factors produced by CD8+ T cells, several chemokine receptors were found to serve as membrane co receptors for primate immunodeficiency lentiretroviruses. The two most widely used co-receptors thus far recognized, CCR5 and CXCR4, are expressed by both activated T lymphocytes and mononuclear phagocytes. CCR5, a specific RANTES, MIP 1alpha and MIP-1 receptor, is used preferentially by non-MT2-tropic HIV-1 and HIV 2 strains and by simian immunodeficiency virus (SIV), whereas CXCR4, a receptor for the C-X-C chemokine SDF-1, is used by MT2-tropic HIV-1 and HIV-2, but not by SIV. Other receptors with a more restricted cellular distribution, such as CCR2b, CCR3 and STRL33, can also function as co-receptors for selected viral isolates. The third variable region (V3) of the gp120 envelope glycoprotein of HIV-1 has been fingered as a critical determinant of the co-receptor choice. Here, we document a consistent pattern of evolution of viral co-receptor usage and sensitivity to chemokine-mediated suppression in a longitudinal follow-up of children with progressive HIV-1 infection. Viral isolates obtained during the asymptomatic stages generally used only CCR5 as a co-receptor and were inhibited by RANTES, MIP-1alpha and MIP-1beta, but not by SDF-1. By contrast, the majority of the isolates derived after the progression of the disease were resistant to C C chemokines, having acquired the ability to use CXCR4 and, in some cases, CCR3, while gradually losing CCR5 usage. Surprisingly, most of these isolates were also insensitive to SDF-1, even when used in combination with RANTES. An early acquisition of CXCR4 usage predicted a poor prognosis. In children who progressed to AIDS without a shift to CXCR4 usage, all the sequential isolates were CCR5 dependent but showed a reduced sensitivity to C-C chemokines. Discrete changes in the V3 domain of gp120 were associated with the loss of sensitivity to C-C chemokines and the shift in co-receptor usage. These results suggest an adaptive evolution of HIV-1 in vivo, leading to escape from the control of the antiviral C C chemokines. PMID- 9359703 TI - A novel protein that participates in nonself discrimination of malignant cells by homologous complement. AB - The human complement (C) system protects an individual against substances of nonself origin, including xenografts and microbial pathogens. Human cells express C-regulatory proteins, CD46 and CD55, thereby circumventing attack by C3, a major effector of C. Nevertheless, certain malignant cells, particularly those undergoing apoptotic stress, can activate homologous C, overcoming the regulatory actions of CD46 and/or CD55. The molecular mechanisms whereby malignant cells are tagged by homologous C3 remain largely unknown. We identified a novel gene product that converts human cells into targets for homologous complement. Only malignant cells and cell lines exposed to Fas or X-irradiation stimuli produced this protein, designated M161Ag, which was an unglycosylated 43-kDa protein. Analysis of cloned cDNAs indicated that this molecule was a secretory protein containing five amino acids encoded by TGA codons. Its functions were unique in that once secreted from the tumor cells, it bound back to the surface of these cells and activated homologous complement (C3) via the alternative pathway, allowing for C3 deposition on the membrane. This molecule may offer new insight into innate immunity; surveillance of tumor cells by complement is a common feature in the human immune system. PMID- 9359704 TI - Evidence for an alternative mechanism for maintaining telomere length in human tumors and tumor-derived cell lines. AB - The gradual loss of DNA from the ends of telomeres has been implicated in the control of cellular proliferative potential. Telomerase is an enzyme that restores telomeric DNA sequences, and expression of its activity was thought to be essential for the immortalization of human cells, both in vitro and in tumor progression in vivo. Telomerase activity has been detected in 50-100% of tumors of different types, but not in most normal adult somatic tissues. It has also been detected in about 70% of human cell lines immortalized in vitro and in all tumor-derived cell lines examined to date. It has previously been shown that in vitro immortalized telomerase-negative cell lines acquire very long and heterogeneous telomeres in association with immortalization presumably via one or more novel telomere-lengthening mechanisms that we refer to as ALT (alternative lengthening of telomeres). Here we report evidence for the presence of ALT in a subset of tumor-derived cell lines and tumors. The maintenance of telomeres by a mechanism other than telomerase, even in a minority of cancers, has major implications for therapeutic uses of telomerase inhibitors. PMID- 9359705 TI - Increased sensitivity to anticancer drugs and decreased inflammatory response in mice lacking the multidrug resistance-associated protein. AB - The multidrug resistance-associated protein (MRP) mediates the cellular excretion of many drugs, glutathione S-conjugates (GS-X) of lipophilic xenobiotics and endogenous cysteinyl leukotrienes. Increased MRP levels in tumor cells can cause multidrug resistance (MDR) by decreasing the intracellular drug concentration. The physiological role or roles of MRP remain ill-defined, however. We have generated MRP-deficient mice by using embryonic stem cell technology. Mice homozygous for the mrp mutant allele, mrp-/-, are viable and fertile, but their response to an inflammatory stimulus is impaired. We attribute this defect to a decreased secretion of leukotriene C4 (LTC4) from leukotriene-synthesizing cells. Moreover, the mrp-/- mice are hypersensitive to the anticancer drug etoposide. The phenotype of mrp-/- mice is consistent with a role for MRP as the main LTC4 exporter in leukotriene-synthesizing cells, and as an important drug exporter in drug-sensitive cells. Our results suggest that this ubiquitous GS-X pump is dispensable in mice, making treatment of MDR with MRP-specific reversal agents potentially feasible. PMID- 9359706 TI - Suppression of a CFTR premature stop mutation in a bronchial epithelial cell line. AB - Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) protein. While 70% of CF chromosomes carry a deletion of the phenylalanine residue 508 (deltaF508) of CFTR, roughly 5% of all CF chromosomes carry a premature stop mutation. We reported that the aminoglycoside antibiotics G-418 and gentamicin can suppress two premature stop mutations [a stop codon in place of glycine residue 542 (G542X) and arginine residue 553 (R553X)] when expressed from a CFTR cDNA in HeLa cells. Suppression resulted in the synthesis of full-length CFTR protein and the appearance of a cAMP-activated anion conductance characteristic of CFTR function. However, it was unclear whether this approach could restore CFTR function in cells expressing mutant forms of CFTR from the nuclear genome. We now report that G-418 and gentamicin are also capable of restoring CFTR expression in a CF bronchial epithelial cell line carrying the CFTR W1282X premature stop mutation (a stop codon in place of tryptophan residue 1282). This conclusion is based on the reappearance of cAMP-activated chloride currents, the restoration of CFTR protein at the apical plasma membrane, and an increase in the abundance of CFTR mRNA levels from the W1282X allele. PMID- 9359707 TI - Osteopetrosis in mice lacking NF-kappaB1 and NF-kappaB2. AB - The nfkb1 and nfkb2 genes encode closely related products regulating immune and inflammatory responses. Their role during development and differentiation remains unclear. The generation of nfkb1 null mice (p50-/-) resulted in altered immune responses, but had no effect on development. Similarly, nfkb2 knockout mice (p52 /-) did not show developmental defects (J.C. et al., manuscript submitted). We have investigated the potential for in vivo compensatory functions of these genes by generating double-knockout mice. The surprising result was that the animals developed osteopetrosis because of a defect in osteoclast differentiation, suggesting redundant functions of NF-kappaB1 and NF-kappaB2 proteins in the development of this cell lineage. The osteopetrotic phenotype was rescued by bone marrow transplantation, indicating that the hematopoietic component was impaired. These results define a new mouse osteopetrotic mutant and implicate NF-kappaB proteins in bone development, raising new directions in the treatment of bone disorders. PMID- 9359708 TI - New developments in the generation of Ad-free, high-titer rAAV gene therapy vectors. PMID- 9359709 TI - Immunotherapy of recurrent genital herpes with recombinant herpes simplex virus type 2 glycoproteins D and B: results of a placebo-controlled vaccine trial. AB - To determine the safety, immunogenicity, and efficacy of a recombinant herpes simplex virus type 2 glycoprotein D and B vaccine in the treatment of recurrent genital herpes, a randomized, placebo-controlled trial was held at two referral centers. Healthy patients with 4-14 recurrences per year received injections of both glycoproteins in MF59 adjuvant or of MF59 alone at 0, 2, 12, and 14 months. For 18 study months, the rate and number of recurrences, the duration and severity of the first confirmed recurrence, vaccine immunogenicity, and rates of local and systemic reactions were determined. The monthly rate of recurrences was not significantly improved, but the duration and severity of the first study outbreak was reduced significantly by vaccination. Glycoprotein-specific and neutralizing antibodies were boosted by vaccination for the duration of the study. This vaccine is safe and immunogenic and ameliorated an observed first postvaccination genital recurrence, but it does not reduce recurrence frequency. PMID- 9359710 TI - Seroconversion to human herpesvirus 6 following liver transplantation is a marker of cytomegalovirus disease. AB - Human herpesvirus 6 (HHV-6) infection is common after transplantation; HHV-6 is known to interact with other viruses and induce immunosuppression. Whether HHV-6 plays a role in the occurrence of cytomegalovirus (CMV) infection after transplantation was investigated. In a cohort of 247 liver transplant recipients, HHV-6 seroconversion was identified as a significant risk factor for development of symptomatic CMV infection (P < .001), including CMV organ involvement (P < .001), even in the presence of the other significant risk factors: D+/R- CMV serologic status (P < .001) or use of OKT3 after transplantation (P = .002). Subgroup analysis indicated that HHV-6 seroconversion was significantly associated with symptomatic CMV infection in the D+/R+ but not in the D+/R- CMV serologic group (P < .001 and P = .11, respectively). These results indicate that HHV-6 seroconversion is a marker for CMV disease after transplantation and suggest that additional studies using more sensitive diagnostic techniques are warranted to determine the relationship between HHV-6 and CMV infection after transplantation. PMID- 9359711 TI - Human papillomavirus type 11 (HPV-11) neutralizing antibodies in the serum and genital mucosal secretions of African green monkeys immunized with HPV-11 virus like particles expressed in yeast. AB - It has been shown previously that immunization of animals with recombinant virus like particles (VLPs) consisting of the viral capsid proteins L1 or L1 plus L2 protected animals against experimental viral challenge. However, none of these experimental models addresses the issue of whether systemic immunization with VLPs elicits a neutralizing antibody response in the genital mucosa. Such a response may be necessary to protect the uterine cervix against infection with genital human papillomavirus (HPV) types. African green monkeys systemically immunized with HPV-11 VLPs expressed in Saccharomyces cerevisiae and formulated on aluminum adjuvant elicited high-titered HPV-11 VLP-specific serum antibody responses. Sera from these immunized monkeys neutralized HPV-11 in the athymic mouse xenograft system. Significant levels of HPV-11-neutralizing antibodies also were observed in cervicovaginal secretions. These findings suggest that protection against HPV infection of the uterine cervix may be possible through systemic immunization with HPV VLPs. PMID- 9359712 TI - Risk for retinitis in patients with AIDS can be assessed by quantitation of threshold levels of cytomegalovirus DNA burden in blood. AB - Cytomegalovirus (CMV) retinitis in patients infected with human immunodeficiency virus (HIV) is a significant clinical problem. Seventy-five patients with CD4 T cell counts <100/mm3 were monitored prospectively every 2 months for CMV DNA burden. The target for DNA amplification was a 162-bp fragment from the CMV immediate early gene. CMV DNA burden, at levels of > or =320 in white blood cells or > or =32 in plasma (P = .001), particularly when sustained (P = .005 and .008, respectively), distinguished patients who developed retinitis from those who remained free of disease. Progression to retinitis was not consistently accompanied by increases in CMV burden, indicating that quantitation of CMV burden beyond threshold levels is not necessary to predict risk for development of retinitis. Virus isolation from WBC, but not urine, was also significantly associated with risk for retinitis (P = .001). PMID- 9359713 TI - Stavudine plus lamivudine in advanced human immunodeficiency virus disease: a short-term pilot study. AB - The short-term effects of stavudine (d4T) plus lamivudine (3TC) were evaluated among 48 human immunodeficiency virus-infected patients for whom zidovudine therapy had failed or who could not tolerate zidovudine. Patients were followed for 8 weeks after initiation of open-label d4T plus 3TC. Four patients discontinued therapy, because of neutropenia (1), hepatitis (1), or neuropathy (2). Reduction in virus load was -0.86 (+0.3 to -3.4) log10 copies/mL and CD4 cell increase was 30 (-100 to +290) cells/mm3. Virologic response was associated with a higher CD4 cell count, no prior exposure to d4T and 3TC, and no previous AIDS-defining illness. Virus load reduction for patients naive to 3TC and d4T was -1.47 (-0.14 to -3.37) log10 copies/mL. Short-term use of d4T plus 3TC is safe, well-tolerated, and associated with virologic and substantial immunologic benefits. Further evaluation of d4T and 3TC in combination is warranted. PMID- 9359714 TI - Predicting clinical progression or death in subjects with early-stage human immunodeficiency virus (HIV) infection: a comparative analysis of quantification of HIV RNA, soluble tumor necrosis factor type II receptors, neopterin, and beta2 microglobulin. Multicenter AIDS Cohort Study. AB - Quantification of human immunodeficiency virus (HIV) RNA by branched-chain DNA signal amplification, measurement of soluble tumor necrosis factor type II receptors (sTNFR-II), neopterin, beta2-microglobulin, or CD4 cell counts can be used to predict the risk of clinical progression or death in HIV infection but have not been compared in the same study. Ninety subjects were categorized into progression groups by their rate of CD4 cell decline and matched into triplets by initial CD4 cell count, age, race, and calendar time. By matched logistic regression, only the sTNFR-II and HIV RNA values were predictive of outcome across the progression groups. Categorization of baseline HIV RNA and sTNFR-II resulted in differences in progression to several clinical outcomes. sTNFR-II concentrations were the only immune marker examined that increased the prognostic utility of HIV RNA determination in early-stage subjects. Further studies in later stages of disease or after therapy are indicated. PMID- 9359715 TI - Chemokine-independent in vitro resistance to human immunodeficiency virus (HIV-1) correlating with low viremia in long-term and recently infected HIV-1-positive persons. AB - Chemokines have been implicated as protective factors against human immunodeficiency virus (HIV) infection, competing for binding to receptors that also function as coreceptors for HIV. In this study of HIV-positive donors, peripheral blood mononuclear cell (PBMC) culture resistance to endogenous and exogenous HIV correlated with low plasma viremia and high in vitro RANTES production. However, resistant cells were not rendered susceptible by neutralization of C-C chemokines, and addition of C-C chemokines did not consistently suppress endogenous virus or exogenous HIV-1MN. In contrast, CD8 T cell depletion markedly decreased the frequency of resistant cultures without reducing C-C chemokine production. Among newly infected persons, half exhibited phenotype switching from preinfection susceptibility to postinfection resistance, suggesting that genetically predetermined constitutive cytokine production or allelic receptor expression are not generally responsible for in vitro resistance and nonprogression. PMID- 9359716 TI - Inhibition of cytokine-driven human immunodeficiency virus type 1 replication by protease inhibitor. AB - Protease inhibitors block virus maturation and prevent the spread of human immunodeficiency virus (HIV)-1 in vitro. HIV-1-positive persons produce higher levels of proinflammatory cytokines that up-regulate HIV-1 replication. For the protease inhibitor to be effective in vivo, it must be able to suppress cytokine induced HIV-1 replication. The in vitro efficacy of protease inhibitor to block tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-1alpha, and IL-1beta induction of HIV-1 was investigated. While 100 U/mL of the respective cytokines induced a 208- to 22-fold increase in HIV-1 p24 production, addition of protease inhibitor completely inhibited this virus induction. The kinetics indicated a sustained HIV-1 inhibition despite high levels of endogenous TNF-alpha induction. Dilution of protease inhibitor led to increased HIV-1 replication. These results show that while protease inhibitor can prevent cytokine induction of HIV-1 replication, a continual effective dose is required for the inhibition to be sustained. PMID- 9359717 TI - Augmented serum neutralizing activity against primary human immunodeficiency virus type 1 (HIV-1) isolates in two groups of HIV-1-infected long-term nonprogressors. AB - Neutralizing activity against primary human immunodeficiency virus type 1 (HIV-1) isolates from 17 persons who were long-term disease nonprogressors (LTNPs) and 13 persons who were fast progressors (FPs) was compared. Sera from LTNPs showed higher neutralizing activity both in titer and in host spectrum than did sera from FPs. However, LTNP sera had limited neutralizing activity against HIV-1 subtypes from different geographic areas. Sera collected 6 years earlier from both groups had limited neutralizing activity, indicating that early responses are not predictive for disease progression. LTNPs had very low virus loads, as reflected by only one positive isolation, which was an MT-2-negative phenotype. Virus was isolated from all FPs, and the isolates showed a phenotype switch from MT-2 negative to MT-2 positive. Development of high-titer, broadly cross-reactive neutralizing antibodies is associated with control of virus replication and low virus load in HIV-1-infected LTNPs. PMID- 9359718 TI - Human immunodeficiency virus type 1 (HIV-1)-specific T cell responses correlate with control of acute HIV-1 infection in macaques. AB - Macaca nemestrina efficiently control acute human immunodeficiency virus type 1 (HIV-1) infection. The roles of helper (Th) and cytotoxic (CTL) T cells in controlling acute HIV-1 infection in both peripheral blood and lymph node mononuclear cells (PBMC and LNMC) were assessed in this model. Th and CTL responses to HIV-1 were detected within 2 weeks following HIV-1 infection, and CTL responses to HIV-1 antigens peaked at 4 weeks after infection (>100 HIV specific CTL/10[6] PBMC), coincident with reductions of HIV-1 RNA and DNA levels in peripheral blood. HIV-1-specific Th and CTL were present in LNMC 6 weeks after infection. Although HIV-1 antibodies were detected 2 weeks after infection, maximal HIV-1 antibody responses were not generated until > 13 weeks after inoculation. Thus, T cell responses temporally correlate with control of HIV-1 in macaques. The induction of a brisk HIV-1-specific CTL response may have been facilitated by a persistent Th response. PMID- 9359719 TI - Simian immunodeficiency virus burden in tissues and cellular compartments during clinical latency and AIDS. AB - In the course of human immunodeficiency virus infection or of the related simian immunodeficiency virus (SIV), progression to AIDS is associated with high virus burdens in blood. How virus burden in the bloodstream is related to virus burden in tissue reservoirs was addressed in an animal model of rhesus macaques infected with SIV. In situ hybridization and quantitative image analysis were used to quantitate virus burden. Animals who developed AIDS had high levels of virus production and storage in lymphoid tissue reservoirs and evidence of productive infection of macrophages in the nervous system. With the quantitative approach described, it should be possible to design and assess the impact of treatment and shed light on the outstanding issues in pathogenesis. PMID- 9359720 TI - Detection of hepatitis C virus RNA in CD19 peripheral blood mononuclear cells of chronically infected patients. AB - The presence of HCV RNA in peripheral blood mononuclear cells (PBMC) has been reported. To identify the cell populations carrying HCV RNA, the presence and amount of HCV RNA was investigated by limiting dilution nested reverse transcriptase-polymerase chain reaction (PCR) in PBMC subpopulations fractionated by automated cell sorting. Fifteen chronically HCV-infected patients were included in the study, 4 of whom also had mixed cryoglobulinemia. HCV RNA was present in the CD19 cells of all 15 patients, but only 5 (35.7%) of 14 and 5 (41.6%) of 12 showed HCV RNA in CD3 and CD14 cells, respectively (P < .001 by Fisher's test for each comparison). The median titer of HCV RNA was 1 PCR unit/380 CD19 cells, compared with median of 1 PCR unit/6600 PBMC as a whole. Titration was difficult in the CD3 and CD14 cells because of the frequent negativity of the first diluted sample. This study suggests that HCV RNA is selectively concentrated in B cells. PMID- 9359721 TI - Development of a humanized monoclonal antibody (MEDI-493) with potent in vitro and in vivo activity against respiratory syncytial virus. AB - Neutralizing polyclonal antibody to respiratory syncytial virus (RSV) has been shown to be an effective prophylactic agent when administered intravenously in high-risk infants. This study describes the generation of a humanized monoclonal antibody, MEDI-493, that recognizes a conserved neutralizing epitope on the F glycoprotein of RSV. The affinity of MEDI-493 was found to be equal to or slightly better than an isotype-matched chimeric derivative of the parent antibody. In plaque reduction, microneutralization, and fusion-inhibition assays, MEDI-493 was significantly more potent than the polyclonal preparation. Broad neutralization of a panel of 57 clinical isolates of the RSV A and B subtypes was demonstrated. Pretreatment of cotton rats with MEDI-493 resulted in 99% reduction of lung RSV titers at a dose of 2.5 mg/kg, corresponding to a serum concentration of 25-30 microg/mL. Further, MEDI-493 did not induce increased RSV infection or pathology in either a primary or a secondary challenge. PMID- 9359722 TI - A randomized evaluation of ethambutol for prevention of relapse and drug resistance during treatment of Mycobacterium avium complex bacteremia with clarithromycin-based combination therapy. California Collaborative Treatment Group. AB - Patients with AIDS and Mycobacterium avium complex (MAC) bacteremia are at high risk for relapse and emergence of resistant isolates during monotherapy with clarithromycin. Ninety-five AIDS patients with MAC bacteremia received clarithromycin plus clofazimine, with or without ethambutol, in a prospective, multicenter, randomized open-label trial. Of 80 patients with positive baseline cultures, sterilization or a 2 log10 reduction in colony-forming units of MAC in two consecutive blood cultures occurred in 69% of both groups. There were nine relapses in the two-drug arm and three in the three-drug arm. Kaplan-Meier estimates of risk of relapse at 36 weeks were 68% and 12%, respectively (P = .004). All relapse isolates were resistant to clarithromycin. Median time to clarithromycin resistance was 16 weeks with two drugs and 40 weeks with three drugs (P = .004). Ethambutol reduced relapses and emergence of clarithromycin resistance and should be considered an essential component of clarithromycin based therapies for MAC bacteremia. PMID- 9359723 TI - Multicenter typing comparison of sporadic and outbreak Clostridium difficile isolates from geographically diverse hospitals. AB - In a collaborative study by three laboratories, arbitrarily primed polymerase chain reaction (AP-PCR), HindIII restriction enzyme analysis (REA), and pulsed field gel electrophoresis (PFGE) using SmaI were compared for typing of Clostridium difficile. The study included 30 isolates from nosocomial outbreaks in six geographically disparate hospitals and 15 isolates from sporadic cases of C. difficile diarrhea. REA distinguished a total of 23 types representing 10 groups; AP-PCR performed at Deaconess Hospital resolved 19 types; AP-PCR performed at the Centers for Disease Control resolved 15 types. Thirty isolates exhibited degradation of larger sized fragments during processing and therefore were nontypeable by PFGE; among the remaining 15 isolates, PFGE resolved 11 types. Outbreak isolates in five different hospitals represented REA group J and constituted a single AP-PCR strain. In summary, nosocomial outbreaks of C. difficile diarrhea in five hospitals were associated with a single genetic lineage as resolved by multiple strain typing systems. PMID- 9359724 TI - The effect of interleukin-10 on meningeal inflammation in experimental bacterial meningitis. AB - Interleukin-10 (IL-10) is a cytokine with antiinflammatory effects. In a rabbit model of meningitis, IL-10 was given intracisternally or intravenously to evaluate the impact on inflammation induced by lipooligosaccharide (LOS), Haemophilus influenzae type b (Hib), or Listeria monocytogenes. Intracisternal IL 10 in concentrations >1 microg significantly reduced tumor necrosis factor-alpha (TNF-alpha) and lactate values in cerebrospinal fluid (CSF). Intravenous IL-10 (1 mg/kg) in two doses after intracisternal LOS significantly reduced CSF TNF-alpha and lactate. When Hib was used, animals were treated with ceftriaxone and dexamethasone with or without IL-10 (1 mg/kg). TNF-alpha was significantly reduced in animals treated with IL-10, dexamethasone, or both compared with levels in rabbits receiving ceftriaxone alone. Comparable results were obtained when L. monocytogenes was inoculated and animals were treated with ampicillin with or without IL-10, dexamethasone, or nothing. In conclusion, IL-10 modulates CSF TNF-alpha concentrations in experimental LOS, Hib, or L. monocytogenes meningitis. The maximal inhibitory effect was seen when IL-10 and dexamethasone were combined. PMID- 9359725 TI - Human immune response to nontypeable Haemophilus influenzae in chronic bronchitis. AB - Nontypeable Haemophilus influenzae (NTHI) causes recurrent respiratory tract infections in patients with chronic bronchitis. To elucidate the human immune response to NTHI, sera from 2 patients with exacerbations of chronic bronchitis due to NTHI were characterized. Both patients developed new bactericidal antibodies following infection. Immunoblot assays with homologous strains revealed antibodies to many antigens, with minimal difference between pre- and postexacerbation sera. By contrast, whole cell radioimmunoprecipitation, which detects antibodies exclusively to epitopes exposed on the bacterial surface, revealed that both patients made new antibodies to a limited number of antigens following infection, including P2, the major outer membrane protein of NTHI. Adsorption experiments showed that strain-specific, surface-exposed epitopes on the P2 molecule are targets for bactericidal antibodies. These results indicate that new bactericidal antibodies following infection by NTHI recognize antigenically heterogeneous surface-exposed epitopes on P2 and other surface proteins of NTHI. PMID- 9359726 TI - Bacteriologic response to oral cephalosporins: are established susceptibility breakpoints appropriate in the case of acute otitis media? AB - Bacteriologic response to cefuroxime axetil and cefaclor administered for 10 days was evaluated in acute otitis media (AOM) in patients aged 6-36 months. Middle ear fluid culture was obtained by tympanocentesis before treatment, on day 4 or 5 after initiation of treatment, and if clinical relapse occurred before day 17. Bacteriologic failure was observed in 32% of patients receiving cefaclor versus 15% of patients receiving cefuroxime axetil (P = .009). Failure rates increased with increasing MIC: For Streptococcus pneumoniae, 0.5 microg/mL (established as cutoff value for cefuroxime by the National Committee for Clinical Laboratory Standards [NCCLS]) discriminated between success and failure. For Haemophilus influenzae, high failure rates were observed for cefaclor, even with low MICs (< or = 1.0 microg/mL), and with both drugs they tended to increase with increasing MIC, even for values below the cutoff suggested by the NCCLS (8.0 and 4.0 microg/mL for cefaclor and cefuroxime, respectively). Thus, for AOM caused by H. influenzae, lower susceptibility cutoff levels for MICs should be established. PMID- 9359727 TI - Antiserum against Escherichia coli J5 contains antibodies reactive with outer membrane proteins of heterologous gram-negative bacteria. AB - The binding of IgG in antiserum to Escherichia coli J5 to the surface of Enterobacteriaceae and to cell wall fragments released from serum-exposed bacteria was studied in a search for potentially protective epitopes other than lipopolysaccharide (LPS). IgG titers to multiple heterologous gram-negative smooth bacteria increased following incubation of the bacteria in serum and decreased following absorption with serum-exposed heterologous bacteria. IgG eluted from absorbing bacteria bound to at least three conserved bacterial outer membrane proteins (OMPs), but not LPS, as assessed by immunoblotting. The same OMPs were present in LPS-containing macromolecular cell wall fragments released by incubation of heterologous gram-negative bacteria in human serum. Part of the protection offered by J5 antiserum could be from binding of IgG to conserved OMPs at the bacterial surface or to OMPs in cell-wall fragments released from dying bacteria. PMID- 9359728 TI - High-level tetracycline-resistant Neisseria gonorrhoeae in Ontario, Canada- investigation of a cluster of isolates, showing chromosomally mediated resistance to penicillin combined with plasmid-mediated resistance to tetracycline. AB - Between 1991 and 1994, plasmid-mediated, tetracycline-resistant Neisseria gonorrhoeae (TRNG) increased from 61.8% to 85.96% of all resistant isolates in Ontario, Canada. Ninety-nine isolates with tetracycline MICs >32 mg/L were characterized by auxotype/serovar (A/S) class, plasmid profile, hybridization with eight tetracycline-resistant probes, and pulsed-field gel electrophoresis (PFGE) of genomic DNA after digestion with NheI and SpeI restriction endonucleases. A cluster of 82 isolates with penicillin MICs of 2-4 mg/L and tetracycline MICs of 128 mg/L (chromosomally mediated resistance) belonged to A/S class NR/IB-1 and had identical or closely related PFGE profiles. Seventeen isolates, TRNG (10) and penicillinase-producing TRNG (7), with tetracycline MICs of 64-256 mg/L, belonged to eight A/S classes and displayed 12 different PFGE profiles. The 99 isolates hybridized only with the TetM probe. Phenotypic and molecular characterization indicated a diverse population throughout the Province of Ontario. PMID- 9359729 TI - Molecular epidemiology of sporadic (endemic) serogroup C meningococcal disease. AB - Understanding the basis of sporadic (endemic) meningococcal disease may be critical to prevention of meningococcal epidemic outbreaks and to understanding fluctuations in incidence. Active, prospective, population-based surveillance and molecular epidemiologic techniques were used to study sporadic serogroup C meningococcal disease in a population of 2.34 million persons (Atlanta area). During 1988-1994, in which no outbreaks or case clusters were reported, 71 patients developed sporadic serogroup C meningococcal disease (annual incidence, 0.51/100,000). Eighty-three percent of patients were >2 years old. By multilocus enzyme electrophoresis, pulsed-field gel electrophoresis, and serotyping, 84% (52/62) of the isolates available for study were identical or closely related members of the electrophoretic type 37 (ET 37) complex responsible for multiple serogroup C outbreaks in the United States in the 1990s. Sporadic disease caused by 9 clonal strains occurred over periods up to 4 years and accounted for 45% (28/62) of cases. Sporadic serogroup C meningococcal disease was most often due to a limited number of related strains that appear to slowly circulate in the population. PMID- 9359730 TI - Interaction of Neisseria maningitidis with the components of the blood-brain barrier correlates with an increased expression of PilC. AB - A fatal untreated case of fulminant meningococcemia was examined to investigate the crossing of the blood-brain barrier (BBB) by Neisseria meningitidis. Microscopic examination showed bacteria in vivo adhering to the endothelium of both the choroid plexus and the meninges. Comparison of the isolates cultivated from the blood and the cerebrospinal fluid (CSF) revealed no antigenic variation of the pilin or the class 5 protein, whereas the expression of the PilC protein was greater in the CSF and the choroid plexus than in the blood. This was due to an increased activity of one of the pilC promotors. This higher expression of PilC correlated in vitro with greater adhesiveness to endothelial cells. A mutation in the single pilC locus of this strain abolished in vitro pilus mediated adhesion to endothelial cells. These data suggest that PilC plays an important role in the crossing of the BBB, likely through pilus-mediated adhesion. PMID- 9359731 TI - Intraportal lipopolysaccharide suppresses pulmonary antibacterial defense mechanisms. AB - Translocation of enteric bacteria or their components (or both) has been postulated to play a role in precipitating sepsis or the systemic inflammatory response syndrome. To simulate the effects of translocation on pulmonary host defenses, lipopolysaccharide was injected into the portal vein of normal rats that were subsequently challenged by aerosol inoculation with Pseudomonas aeruginosa. Injection of LPS into the portal vein resulted in increased serum tumor necrosis factor (TNF)-alpha levels and reduction in lung clearance of P. aeruginosa after aerosol challenge. There were corresponding reductions in alveolar neutrophil recruitment, diminished alveolar macrophage phagocytosis and superoxide anion (O2-) production, and diminished lung TNF recovered by bronchoalveolar lavage. Furthermore, prior intravenous injection of recombinant TNF-alpha reproduced the defective bacterial clearance, the altered recruitment of airspace neutrophils, and the defective alveolar macrophage phagocytosis. Thus, systemic TNF-alpha is important in altering pulmonary defenses, and this work supports the concept that bacterial translocation may adversely affect host defenses in distant organs. PMID- 9359732 TI - Targeted suppression of cytokine production in monocytes but not in T lymphocytes by a tetravalent guanylhydrazone (CNI-1493). AB - Image analysis was used to study the cytokine-inhibitory effect of the nitric oxide inhibitor tetravalent guanylhydrazone (CNI-1493) in individual immunocytochemically stained human peripheral blood mononuclear cells (PBMC). CNI 1493 inhibited lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) alpha, interleukin (IL)-1alpha, IL-1beta, IL-6, and IL-8 production whether or not LPS stimulation was enhanced by interferon (IFN)-gamma priming. Addition of TNF-alpha to CNI-1493-exposed LPS-stimulated cells partially restored the incidence of IL-1alpha-, IL-1beta-, and IL-8-producing cells. TNF-alpha production induced by costimulation by ligation of CD3 and CD28 was inhibited by CNI-1493 in monocytes but not in T lymphocytes. The prevalence of IL-2-, IFN gamma-, and TNF-beta-producing T cells was not reduced by CNI-1493. Phorbol ester and ionomycin activation also resulted in a CNI-1493 -induced inhibition of TNF alpha in monocytes but resistant production of TNF-alpha, IL-2, and IFN-gamma by T cells. Thus, CNI-1493 preferentially inhibited synthesis of proinflammatory cytokines in monocytes. PMID- 9359733 TI - Relationship of tissue and cellular interleukin-1 and lipopolysaccharide after endotoxemia and bacteremia. AB - Distributions of immunoreactive interleukin-1 (IL-1) and lipopolysaccharide (LPS) were studied in the tissues of rats after intravenous injection of purified LPS or live Escherichia coli bacteria. IL-1 staining in the spleen peaked at 4-8 h, colocalized with LPS in marginal zone macrophages, and was undetectable 24 h after injection, whereas LPS staining peaked at 24 h and was detectable for 4 weeks. The tissue IL-1 response was similar for LPS and live bacteria. Thus, tissue IL-1 is down-regulated within hours despite maintenance of LPS in the same cells for weeks. Macrophages in liver and lung had only slight IL-1 staining despite intense staining for LPS. Tissue IL-1 production appears to be differentially regulated after gram-negative bacteremia; LPS cleared by liver and lung macrophages elicit minimal IL-1, whereas there is high local IL-1 production in the marginal zone of the spleen that may increase immune responses to bacterial wall antigens. PMID- 9359734 TI - Pneumocystis carinii host origin defines the antibody specificity and protective response induced by immunization. AB - To determine how the known host species-specific antigenic variation of Pneumocystis carinii would affect immune recognition, mice were immunized with either mouse- or ferret-derived P. carinii, with subsequent analysis of the immune response and the ability of the mice to resist infection after immunosuppression and challenge. Immunization with mouse-derived P. carinii produced a strong immune response to mouse but not ferret P. carinii. These mice were completely protected from P. carinii pneumonia when challenged by intratracheal inoculation with mouse P. carinii. In contrast, immunization with ferret P. carinii produced a limited antibody response to mouse P. carinii and had no protective effect. These results show that P. carinii from different host species are immunologically distinct, and any possible use of immunotherapy for P. carinii pneumonia in humans must take into consideration these biologically significant antigenic differences in P. carinii of animal origin. PMID- 9359735 TI - The novel oxygenated chalcone, 2,4-dimethoxy-4'-butoxychalcone, exhibits potent activity against human malaria parasite Plasmodium falciparum in vitro and rodent parasites Plasmodium berghei and Plasmodium yoelii in vivo. AB - Previous studies have shown that licochalcone A, an oxygenated chalcone, exhibits antileishmanial and antimalarial activities. The present study was designed to examine the antimalarial activity of an analog of licochalcone A, 2,4-dimethoxy 4'-butoxychalcone (2,4mbc). 2,4mbc inhibited the in vitro growth of both a chloroquine-susceptible (3D7) and a chloroquine-resistant (Dd2) strain of Plasmodium falciparum in a [3H]hypoxanthine uptake assay. The in vivo activity of 2,4mbc was tested in mice infected with Plasmodium berghei or Plasmodium yoelii and in rats infected with P. berghei. 2,4mbc administered either orally, intraperitoneally, or subcutaneously for 5 days protected the mice from otherwise lethal infections of these parasites. 2,4mbc administered orally for 5 days reduced parasitemia in the rats infected with P. berghei. These results demonstrate that 2,4mbc exhibits potent antimalarial activity and might be developed into a new antimalarial drug. PMID- 9359736 TI - Immunization with the cross-reactive antigens Ov39 from Onchocerca volvulus and hr44 from human retinal tissue induces ocular pathology and activates retinal microglia. AB - Antigen Ov39, derived from Onchocerca volvulus, cross-reacts on both the T and B cell level with a nonhomologous human retinal antigen, hr44. Lewis rats were immunized to investigate the potential of these antigens to induce eye disease. Histologic and immunohistologic examination of ocular tissues revealed pathologic changes as early as day 12, which included induction or up-regulation of class II and CD68-like antigen on perivascular cells, ramified retinal microglia, dendritiform cells of the iris epithelium, and ciliary epithelium and significant breakdown of anterior and posterior blood-ocular barriers. Extravascular immunoglobulin and staining for CD68-like antigen was detected in the optic nerve after immunization with Ov39. Unrelated structural abnormalities of retina and lens seen in 8% of eyes examined significantly predisposed eyes to the development of Ov39- or hr44-induced pathology. These findings suggest a role for cross-reactive immune responses in the development of ocular onchocerciasis. PMID- 9359737 TI - Anti-schistosome IgG4 and IgE at 2 years after chemotherapy: infected versus uninfected individuals. AB - Specific IgG4 and IgE responses to adult worm antigen (AWA) and soluble egg antigen (SEA) were examined in 37 subjects from an area in which schistosomiasis is endemic, who were previously infected with Schistosoma haematobium and who became reinfected or remained free of infection 2 years after chemotherapy. The reinfected group was significantly younger (median age, 11 years) than the uninfected group (median age, 24 years). Posttreatment levels of IgG4 to egg antigens (IgG4-SEA) were significantly correlated with reinfection intensity (r = .74, P < .0001), and 13-fold lower levels of IgG4-SEA were observed in uninfected subjects compared with reinfected subjects. Although no correlation was observed between posttreatment IgE to AWA or to SEA, pretreatment IgE-AWA was inversely correlated with the level of reinfection (r = -.39, P = .02). PMID- 9359738 TI - Review: the immune response in human tuberculosis--implications for tuberculosis control. PMID- 9359739 TI - Low detection rate and maternal provenance of hepatitis B virus S gene mutants in cases of failed postnatal immunoprophylaxis in England and Wales. AB - Hepatitis B virus (HBV) infection occurred despite full passive-active immunoprophylaxis in 20 of 321 infants born to mothers seropositive for hepatitis B e antigen. In 2 (12%) of 17 infected infants, mother-infant DNA sequence mismatches were found in a segment of the HBV S gene coding for antigenic determinants of the HBV surface antigen (HBsAg) amplified from sera by polymerase chain reaction (PCR). Point substitutions occurred in codons 120, 134, and 144 of the HBsAg polypeptide in the variant sequence of 1 infant and in codon 126 in the other; all were missense mutations. Mutant sequences could not be recovered from maternal sera by PCR cloning but were selectively generated using an amplification refractory mutation system. The frequency of potential vaccine escape mutants is therefore low, and these preexist maternally as minor variants. PMID- 9359740 TI - Short-term safety of live attenuated Japanese encephalitis vaccine (SA14-14-2): results of a randomized trial with 26,239 subjects. AB - The short-term safety of an effective and inexpensive new live attenuated Japanese encephalitis vaccine (SA14-14-2) was studied in a randomized trial, using block randomization. Of 26,239 children who were enrolled, half received the vaccine and half served as controls. Subjects were prospectively followed for 30 days for severe adverse events, such as encephalitis, meningitis, and "all cause" hospitalization. No cases of encephalitis or meningitis occurred in either group. The upper 95% confidence limit for adverse events not occurring among subjects receiving their first dose was 4.1/10,000. Risk ratios and 95% confidence intervals for other adverse events were 0.70 (0.43-1.15) for all-cause hospitalization, 0.91 (0.37-2.22) for seizure, and 0.79 (0.56-1.11) for fever lasting > or = 3 days. These data attest to the short-term safety of the SA14-14 2 virus strain and the hamster kidney cell substrate. PMID- 9359741 TI - The 30-bp deletion variant of Epstein-Barr virus-encoded latent membrane protein 1 prevails in acute infectious mononucleosis. AB - To assess the frequency of malignancy-associated 30-bp deletion variants of the latent membrane protein 1 (LMP-1) in benign conditions, a comparative sequence analysis was done using samples from 20 American children with acute infectious mononucleosis and 16 Swiss children with chronic tonsillar hyperplasia. The 30-bp deletion variant (LMP-1-del) was present in 66% of patients (12/20 with infectious mononucleosis and 12/16 with tonsillar hyperplasia). Two additional patients had a 3-bp deletion and an inframe insertion of 18 nucleotides, respectively. All but 1 isolate had numerous nonsilent point mutations. These data identify a hypervariable region within the C-terminus of LMP-1, in a domain required for maximal stimulation of NF-kappaB activity. These data demonstrate that LMP-1-del variants are frequent in acute infectious mononucleosis and tonsillar hyperplasia and identical to those observed in Epstein-Barr virus associated AIDS-related lymphoma. PMID- 9359742 TI - The incidence and genetic variability of small round-structured viruses in outbreaks of gastroenteritis in The Netherlands. AB - Small round-structured viruses (SRSV) are a group of RNA viruses that can cause gastroenteritis in persons of all ages. To determine the incidence of SRSV associated gastroenteritis in The Netherlands and to study the genetic variability of outbreak strains, all outbreaks that were reported to the epidemiologists of the regional health services in 1996 were investigated using a standardized protocol. In 60 (87%) of the 69 reported outbreaks, SRSV could be detected, showing the etiologic significance of SRSV in outbreaks of gastroenteritis in The Netherlands. Of these outbreaks, 84% occurred in semiclosed communities, such as nursing homes (59%) and hospital wards (25%). Sequence analysis of the outbreak strains revealed that the majority of the strains from January to November 1996 formed a tight cluster within genogroup II SRSV. In November 1996, a shift toward genogroup I SRSV occurred, suggesting a change to a new predominant strain. PMID- 9359743 TI - Lymph node expansion of CD4+ lymphocytes during antiretroviral therapy. AB - The evolution of lymphocyte subsets was analyzed in sequential lymph nodes (LN) biopsies and compared with that in the blood of 25 human immunodeficiency virus type 1 (HIV-1)-infected patients receiving highly active antiretroviral therapy. An average of 3 biopsies were obtained from each patient, with a mean follow-up of 5.6 +/- 0.6 months. A correlation was found between the CD4:CD8 ratio in blood and in LN at baseline but not after > or = 2 months of therapy. With therapy, there was a significant increase in CD2+ cells and a much higher CD4+ cell increase and CD8+ cell decrease in LNs compared with levels in blood. A subset of patients had increased expression of Ki-67 and a decreased expression of CD8CD38 or CD3HLA-DR. Expanded CD4+ cells in LNs were mainly CD45RO+, and changes were concomitant with a decrease in LN virus load. These data demonstrate that CD4 cell reconstitution in HIV-1 infection takes place primarily in secondary lymphoid organs and is not related to a simple redistribution of cells. PMID- 9359744 TI - Temporal relationship between human immunodeficiency virus type 1 RNA levels in serum and cellular infectious load in peripheral blood. AB - Cross-sectional analysis of 252 paired serum and peripheral blood mononuclear cell (PBMC) samples derived from 54 human immunodeficiency virus type 1 (HIV-1) infected persons revealed a correlation between HIV-1 RNA load in serum and infectious load in peripheral CD4 T cells after 18 months of follow-up and before an AIDS diagnosis (Pearson's correlation coefficient [r(p)] = .71, P < .001) and during antiviral treatment (r[p] = .78, P < .001). To gain insight into the temporal relationship between both measures of virus load, longitudinally obtained samples from 23 persons with various clinical courses (slow or rapid disease progression, long-term survival) and 22 persons undergoing antiviral therapy (zidovudine or didanosine, or both, or ritonavir) were analyzed. In general, the kinetics of changes in both measures of virus load were similar in the natural course of infection (78% of study participants) and during treatment (82% of participants). These findings suggest that PBMC and serum represent closely related, if not the same, viral compartments. PMID- 9359745 TI - Correlation of virus load in plasma and lymph node tissue in human immunodeficiency virus infection. INCAS Study Group. Italy, Netherlands, Canada, Australia, and (United) States. AB - The impact of long-term changes in plasma viremia, produced by effective combination antiretroviral therapy, on human immunodeficiency virus (HIV) burden within tissue reservoirs is unknown. Fifteen patients who had received at least 1 year of therapy with two or three drug combinations of zidovudine, didanosine, and nevirapine had suitable samples of lymph node tissue obtained by ultrasound guided core needle biopsy. HIV RNA was extracted from homogenized tissue samples and quantitated using a modified branched DNA assay. Results were correlated with antiretroviral treatment effect on the basis of plasma virus load measurements over the preceding 12-18 months. A statistically significant negative correlation was observed between magnitude of treatment effect on plasma viremia and lymph node virus load. These data suggest that combinations of antiretroviral drugs that produce sustained suppression of plasma HIV RNA may also be able to reduce the virus burden in lymphoid tissues. PMID- 9359746 TI - Zidovudine-resistant human immunodeficiency virus type 1 strains subcultured in the presence of both lamivudine and quinoxaline HBY 097 retain marked sensitivity to HBY 097 but not to lamivudine. AB - Replication of zidovudine-resistant human immunodeficiency virus type 1 (HIV-1) strains (containing the 41 Met-->Leu and 215 Thr-->Tyr mutations in reverse transcriptase [RT]) was inhibited to a significantly greater extent by the combination of lamivudine and quinoxaline HBY 097 than by either drug alone or even fully suppressed by concomitant HBY 097 and lamivudine administration at relatively low concentrations. The virus recovered after exposure to the drug combinations individually had acquired the 103 Lys-->Arg, 138 Glu-->Lys, 184 Met- >Ile, and 189 Val-->Ile mutations in the genetic zidovudine-resistance background of zidovudine-resistant HIV-1. These mutants retained marked sensitivity to HBY 097. The genotypic zidovudine-resistance mutations were maintained in the mutant virus RT genomes, and the viruses also remained phenotypically resistant to zidovudine. Given the exquisite potency of the combination of lamivudine and HBY 097 in suppressing viral replication, this combination should be further pursued in clinical trials examining treatment of HIV-1-infected persons. PMID- 9359747 TI - Syphilis serology in human immunodeficiency virus infection: evidence for false negative fluorescent treponemal testing. AB - Injection drug users were assessed serologically for human immunodeficiency virus infection and syphilis every 6 months. Treatment histories were reviewed for any high-titer biologic false-positive (BFP) reactors, that is, persons with rapid plasma reagin (RPR) titers > or = 1:4 and negative results for fluorescent treponemal antibody absorption (FTA-ABS) tests. Selected sera were analyzed further by immunoblotting for the presence of antibodies reactive with specific Treponema pallidum antigens. Of 112 BFP reactors, 35 (31%) had at least one RPR test reactive at a dilution >1:8 while the FTA-ABS test remained nonreactive. Five reactors (4.5%) converted from nonreactive to reactive by FTA-ABS test; 4 (3.6%) were reactive by FTA-ABS tests but later became nonreactive. Antibodies to T. pallidum membrane antigens were detected in some samples that were persistently nonreactive by FTA-ABS test. Serologic patterns over time, along with very high-titer BFP reactions and reactivity with T. pallidum-specific antigens, suggest that some BFP reactions may represent FTA-negative syphilis. PMID- 9359749 TI - Primary bone marrow progenitors of both granulocytic and monocytic lineages are susceptible to infection with the agent of human granulocytic ehrlichiosis. AB - Human granulocytic ehrlichiosis (HGE) is an emerging tickborne infection resulting in an acute febrile illness associated with cytopenias and characteristic intracellular organisms within peripheral blood granulocytes. The etiologic agent of HGE has recently been isolated and cultivated in the HL-60 promyelocytic leukemia cell line, but the spectrum of host cells that it naturally infects remains unknown. To determine if normal hematopoietic progenitors could be targets of infection, CD34+ primary human bone marrow cells, stimulated to differentiate along myelomonocytic lineages, were incubated with the HGE agent. Immature marrow progenitors and, remarkably, not only granulocytic but also CD14+ monocytic cells from these cultures supported replication of the HGE agent, suggesting that all are potential targets of infection in vivo. Infection of bone marrow progenitors may contribute to the hematologic manifestations of HGE. Furthermore, the ability of the agent to interact with monocytes has significant implications regarding disease pathogenesis and host response. PMID- 9359748 TI - BOX-polymerase chain reaction-based DNA analysis of nonserotypeable Streptococcus pneumoniae implicated in outbreaks of conjunctivitis. AB - Nonserotypeable isolates predominate in epidemic conjunctivitis caused by Streptococcus pneumoniae. Previous evaluations of outbreaks of pneumococcal conjunctivitis have relied on epidemiologic factors and the nontypeability of the isolates to infer that a single clone was involved. In the present study, BOX polymerase chain reaction DNA analysis was used to characterize nonserotypeable S. pneumoniae isolated by conjunctival culture during a recent conjunctivitis outbreak and to compare these isolates with those from outbreaks described earlier. The recent outbreak was caused by a single pneumococcal clone. Outbreaks in separate parts of the United States in 1980-1981 were all caused by the same clone. Cluster analysis revealed a high degree of genetic relatedness among isolates causing conjunctivitis compared with that among other nonserotypeable S. pneumoniae, with the closest relatedness being found among the 1996 and 1980-1981 conjunctival isolates. PMID- 9359750 TI - Neutralization assays using the BZ167 strain of human immunodeficiency virus type 1. PMID- 9359751 TI - Does prior tuberculosis protect human immunodeficiency virus-infected persons from Mycobacterium avium complex disease?